PMID- 11378237 TI - Which synapses are affected in aging and what is the nature of their vulnerability? A commentary on "life span and synapses: will there be a primary senile dementia?". PMID- 11378238 TI - Synaptic loss and pathological change in older adults--aging versus disease? PMID- 11378240 TI - Quantitative assessment of possible age-related change in synaptic numbers in the human frontal cortex. AB - To investigate possible age-associated changes in human synaptic connectivity, superior-middle frontal cortex (Brodmann area 9) was evaluated with ultrastructural techniques. Short post mortem autopsy tissue was obtained from 37 cognitive normal individuals ranging in age from 20 to 89 years. A minimum of five subjects represented each decade of life. Synaptic volume density (Nv) was quantified in lamina III and V of the superior-middle frontal cortex employing the physical disector. The stereological assessment demonstrated maintenance of Nv in both lamina III and V of the frontal cortex. The lack of synaptic decline in the frontal cortex in neurologically normal individuals older than 65 years lends support to the idea that many stereotypic views of age-related changes in the CNS do not apply to all brain regions. It also suggests that synaptic loss observed in pathological conditions such as Alzheimer's disease, may be the result of the disease process and not a consequence of normal aging. PMID- 11378241 TI - Distinguishable effects of presenilin-1 and APP717 mutations on amyloid plaque deposition. AB - Both APP and PS-1 are causal genes for early-onset familial Alzheimer's disease (AD) and their mutation effects on cerebral Abeta deposition in the senile plaques were examined in human brains of 29 familial AD (23 PS-1, 6 APP) cases and 14 sporadic AD cases in terms of Abeta40 and Abeta42. Abeta isoform data were evaluated using repeated measures analysis of variance which adjusted for within subject measurement variation and confounding effects of individual APP and PS-1 mutations, age at onset, duration of illness and APOE genotype. We observed that mutations in both APP and PS-1 were associated with a significant increase of Abeta42 in plaques as been documented previously. In comparison to sporadic AD cases, both APP717 and PS-1 mutation cases had an increased density (measured as the number of plaques/mm(2)) and area (%) of Abeta42 plaques. However, we found an unexpected differential effect of PS-1 but not APP717 mutation cases. At least some of PS-1 but not APP717 mutation cases had the significant increase of density and area of Abeta40-plaques as compared to sporadic AD independently of APOE genotype. Our results suggest that PS-1 mutations affect cerebral accumulation of Abeta burden in a different fashion from APP717 mutations in their familial AD brains. PMID- 11378242 TI - Correlation between cognitive deficits and Abeta deposits in transgenic APP+PS1 mice. AB - Doubly transgenic mAPP+mPS1 mice (15-16 months) had impaired cognitive function in a spatial learning and memory task that combined features of a water maze and a radial arm maze. Nontransgenic mice learned a new platform location each day during 4 consecutive acquisition trials, and exhibited memory for this location in a retention trial administered 30 min later. In contrast, transgenic mice were, on average, unable to improve their performance in finding the hidden platform over trials. The cognitive performance of individual mice within the transgenic group were inversely related to the amount of Abeta deposited in the frontal cortex and hippocampus. These findings imply that mAPP+mPS1 transgenic mice develop deficits in cognitive ability as Abeta deposits increase. These data argue that radial arm water maze testing of doubly transgenic mice may be a useful behavioral endpoint in evaluating the functional consequences of potential AD therapies, especially those designed to reduce Abeta load. PMID- 11378243 TI - Calreticulin functions as a molecular chaperone for the beta-amyloid precursor protein. AB - Processing of the beta-amyloid precursor protein (APP) in the endoplasmic reticulum and the Golgi apparatus may be critical in generating the beta-amyloid molecules linked to the pathogenesis of Alzheimer's disease. Since chaperone molecules such as calreticulin (Crt) have been shown to be important in the maturation of many glycoproteins, we investigated the interaction between Crt and APP. We show that APP binds transiently to Crt in a manner that is pH, divalent cation, and N-linked glycosylation-dependent. Both immature APP (containing only N-linked sugars) and mature APP (containing both N-linked and O-linked sugars) bind to Crt. Both proteins are part of a complex that appears to be large enough to accommodate other proteins as well. However, while most of the immature form is associated with the complexes, very little of the mature form is. The interaction between APP and Crt is likely to be of physiological significance with respect to APP maturation since Crt is involved in quality control of nascent glycoproteins in the secretory pathway. PMID- 11378244 TI - Increased protein glycation in cerebrospinal fluid of Alzheimer's disease. AB - Accumulation of advanced glycation end products occurs in the brain with ageing and was proposed to be involved in pathogenesis of Alzheimer's disease. We studied changes in the level of an early glycation product, an Amadori product, in cerebrospinal fluid (CSF) in ageing and in late-onset Alzheimer's disease. The work was carried out on 99 consecutive patients. The concentration of Amadori product in CSF correlated with CSF glucose concentration but was not changed with age (n = 70). In contrast, level of CSF Amadori product was 1.7-fold higher in Alzheimer's disease patients (n = 29) as compared with non-demented age-matched control group (n = 20; P < 0.0005), although CSF glucose concentration was similar in both groups (4.1 +/- 1.3 vs. 3.8 +/- 0.6 mmol/liter, resp.). An increased accumulation of Amadori products was found in all major proteins of CSF of Alzheimer's disease including albumin, apolipoprotein E and transthyretin. We propose that the increased early glycation of CSF proteins in the Alzheimer's patients may stimulate the formation and the consequent deposition of advanced glycation end products as well as oxidative stress in the brain. PMID- 11378245 TI - N(epsilon)(gamma-glutamyl)lysine in cerebrospinal fluid marks Alzheimer type and vascular dementia. AB - N(epsilon)(gamma-glutamyl)lysine isodipeptide is released from the breakdown of proteins cross-linked by transglutaminase enzymes. Transglutaminase activation is a marker of apoptosis and elevated isodipeptide concentrations in body fluids might correlate with the intensity of apoptotic cell turnover. The concentration of N(epsilon)(gamma-glutamyl)lysine was measured in the cerebrospinal fluid (CSF) of patients with probable Alzheimer's disease (n = 14) and vascular type dementia (n = 11) and compared with not demented surgical controls (n = 17). Baseline levels of 26-62 nM/l (mean 37.9 +/- 8.7 SD) free isodipeptide were detected in control patients. CSF isodipeptide levels showed significant elevation in vascular (mean 95.6 +/- 45.1 SD) as well as Alzheimer patients (176.6 +/- 77.1 SD). Isodipeptide concentrations above 120 nM/l were 72% specific and 77% sensitive to Alzheimer's dementia, although the difference between the two dementias was statistically insignificant (p > 0.05). Determination of CSF N(epsilon)(gamma-glutamyl)lysine isodipeptide concentration offers a novel method for measurement of neurodegeneration in primary and mixed dementias. PMID- 11378246 TI - Antihypertensive drugs and incidence of dementia: the Rotterdam Study. AB - There is increasing evidence that hypertension may contribute to the development of dementia. We investigated the relation of antihypertensive drug use and the risk of dementia in the cohort of the population based Rotterdam Study. The study cohort included 7046 elderly, free of dementia at baseline. Dementia was diagnosed in a stepwise procedure. Participants were first screened. Screen positives were further tested. Those suspected of dementia underwent a diagnostic work-up. Dementia and its subtypes were diagnosed according to prevailing criteria. A Cox proportional hazards model was used to estimate relative risks. After a mean follow-up of 2.2 years, subjects taking antihypertensive medication at baseline (n = 2015) had a reduced incidence of dementia (adjusted relative risk, 0.76; 95% confidence interval 0.52-1.12). This risk reduction was most pronounced for vascular dementia, (adjusted relative risk, 0.30; 95% confidence interval 0.11-0.99). For Alzheimer's disease the relative risk was 0.87, not significant. Dementia may be prevented by antihypertensive treatment. In order to confirm any relation in Alzheimer's disease larger observational studies with longer follow-up are needed. PMID- 11378247 TI - Plasma alpha1-antichymotrypsin in Alzheimer's disease; relationships with APOE genotypes. AB - Inflammatory processes are thought to be important contributors to the pathogenesis of Alzheimer's disease (AD). alpha1-antichymotrypsin (ACT) is a proteinase inhibitor characteristic of acute-phase inflammation and has been identified in amyloid plaques. We analyzed the plasma ACT levels in a sample of subjects with late-onset AD and correspondent controls. Plasma ACT was higher in AD patients (62.8 +/- 20.2 mg/dl) than in controls (58.8 +/- 18.1 mg/dl), but not significantly (P = 0.13). In the AD patients regression analysis showed a positive linear relationship between ACT levels and duration of the disease (P = 0.037). Increased ACT concentrations (64.6 +/- 21.2 mg/dl) were also found in patients with greater cognitive impairment (MMSE scores < 20), but since this factor depends on the duration of the disease as well, our present data seem to indicate a complex relationship involving elevated ACT levels, disease duration and cognitive impairment. Plasma ACT was found to differ significantly according to APOE genotypes (P = 0.017), the highest levels being associated to E3-E3 homozygotes (66.1 +/- 17.8 mg/dl) and the lowest to E4-E3 subjects (53.1 +/- 18.2 mg/dl). In patients not carrying APOE*4 allele the ACT levels were higher than in controls (P = 0.014), and the relationship between ACT and disease duration was stronger than that observed in the total AD sample (P = 0.003), but it was absent in those carrying APOE*4 (P = 0.67). Taken together our results seem to suggest that inflammation is a relevant factor in AD pathogenesis for subjects with E3-E3 and E3-E2 genotypes but less important for APOE*4 carrying subjects. PMID- 11378248 TI - Increased expression of estrogen receptor alpha and beta in the nucleus basalis of Meynert in Alzheimer's disease. AB - The human nucleus basalis of Meynert (NBM) is severely affected in Alzheimer's disease (AD). Since estrogens may reduce both the risk and severity of AD, possibly by an action on the cholinergic system, we determined whether estrogen receptors are present in the human NBM and what their changes are in normal aging and in AD. ERalpha was expressed to a higher degree than ERbeta and was localized mainly in the cell nucleus, while ERbeta was mainly confined to the cytoplasm. A significant positive correlation between the percentage of ERalpha nuclear positive neurons and age was found in men but not in women, whereas the proportion of ERbeta cytoplasm positive cells increased during aging in both sexes. In AD the proportion of neurons showing nuclear staining for both ERalpha and beta and cytoplasmic staining for ERbeta was markedly increased. The percentage of ERbeta nuclear positive neurons increased in AD only in women but not in men. The ApoE genotype had no effect on ER expression in the NBM in AD. In conclusion, whereas only minor sex- and age-related changes in both ERs were found in the human NBM, a clear upregulation of ERalpha and beta was observed in AD. PMID- 11378250 TI - Depressed baroreflex sensitivity in patients with Alzheimer's and Parkinson's disease. AB - Parkinson's disease (PD) and Alzheimer's dementia (AD) are often associated with an autonomic neuropathy. The extent of autonomic involvement, however is poorly defined and unpredictable. In order to assess the autonomic cardiovascular regulation baroreflex sensitivity (BRS) was determined non-invasively in 23 patients (age: 65 +/- 9.3 years) with PD and 24 patients with AD (age: 72.3 +/- 7.2 years). The results were compared with those on 22 healthy age- and sex matched volunteers. Patients with PD and AD exhibited marked abnormalities in cardiovascular autonomic reflex regulation showed by markedly depressed BRS. The possible predictive value of centrally based depression of baroreflex sensitivity necessitates further studies. PMID- 11378249 TI - 3D-Reconstruction of microglia and amyloid in APP23 transgenic mice: no evidence of intracellular amyloid. AB - Microglia cells are closely associated with compact amyloid plaques in Alzheimer's disease (AD) brains. Although activated microglia seem to play a central role in the pathogenesis of AD, mechanisms of microglial activation by beta-amyloid as well as the nature of interaction between amyloid and microglia remain poorly understood. We previously reported a close morphological association between activated microglia and congophilic amyloid plaques in the brains of APP23 transgenic mice at both the light and electron microscopic levels [25]. In the present study, we have further examined the structural relationship between microglia and amyloid deposits by using postembedding immunogold labeling, serial ultrathin sectioning, and 3-dimensional reconstruction. Although bundles of immunogold-labeled amyloid fibrils were completely engulfed by microglial cytoplasm on single sections, serial ultrathin sectioning and three dimensional reconstruction revealed that these amyloid fibrils are connected to extracellular amyloid deposits. These data demonstrate that extracellular amyloid fibrils form a myriad of finger-like channels with the widely branched microglial cytoplasm. We conclude that in APP23 mice a role of microglia in amyloid phagocytosis and intracellular production of amyloid is unlikely. PMID- 11378251 TI - Apolipoprotein E epsilon4 positive individuals demonstrate delayed olfactory event-related potentials. AB - Apolipoprotein E epsilon4 positive individuals have deficits in the ability to identify and remember odors, as demonstrated by psychophysical measures of olfactory function. The purpose of the present study was to identify olfactory deficits in this population using an objective electrophysiological measure: the olfactory event-related potential (OERP). Olfactory and auditory ERPs were recorded from the Fz, Cz, and Pz electrode sites in 10 epsilon4 positive individuals and 10 age and gender-matched epsilon4 negative individuals in a single-stimulus paradigm. The results indicate: (1) individuals who are positive for the apolipoprotein E epsilon4 allele demonstrated delays (of approximately 100 ms) in the processing of olfactory information compared to those who are epsilon4 negative; (2) OERP latency is more sensitive than psychophysical measures of olfactory function; (3) delays in the cognitive P3 component of the OERP are associated with deficits in the ability to identify an odor; and (4) unlike the moderate ranges found in auditory ERP, OERP latency showed high sensitivity and specificity in classifying epsilon4 positive and negative individuals. PMID- 11378253 TI - Effects of a 25-h sleep deprivation on daytime sleep in the middle-aged. AB - Our understanding of the mechanisms by which sleep deteriorates with age almost exclusively stems from comparisons of young and elderly subjects. The present study investigated the different effects of a 25-h sleep deprivation on the recovery sleep initiated in the morning (when circadian sleep propensity decreases) of young (20-39 y) and middle-aged subjects (40-60 y). Middle-aged subjects showed a steeper increase in the duration of wakefulness during daytime recovery sleep than the young subjects. Slow-wave sleep (SWS) and EEG slow-wave activity (SWA: spectral power between 0.5-4.5 Hz) were potentiated in both groups following sleep deprivation. However, the rebound of SWS and SWA was significantly less pronounced in the middle-aged than in the young. This reduction in homeostatic recuperative drive in middle-aged subjects might account for the decrease in their ability to maintain sleep when they have to recuperate at an abnormal circadian phase. These results helps to understand the increase in complaints related to shift work and jet lag in the middle years of life. PMID- 11378252 TI - MRI and genetic correlates of cognitive function in elders with memory impairment. AB - The present study investigated the relationship between genetic variation, MRI measurements and neuropsychological function in a sample of 58 elders exhibiting memory decline. In agreement with previous reports, we found that the epsilon4 allele of the apolipoprotein E (APOE) and the D allele of the angiotensin converting enzyme (ACE) polymorphisms negatively modulated the cognitive performance. Further, we found an association between the A allele of the apolipoprotein C1 (APOC1) polymorphism and poorer memory and frontal lobe function. No clear associations emerged between MRI measures of white matter lesions (WML) or hippocampal sulcal cavities (HSC) and the cognitive performance after controlling for age effects. Further, the degree of WML or HSC lesions was in general not predisposed genetically except for the presence of the A allele of the APOC1 polymorphism that was related to a higher severity of HSC scores. Our results suggest that WML or HSC do not represent important brain correlates of genetic influences on cognitive performance in memory impaired subjects. PMID- 11378254 TI - Ultrastructural and permeability features of microvessels in the hippocampus, cerebellum and pons of senescence-accelerated mice (SAM). AB - We previously reported that the accumulation of blood-borne radiolabelled serum albumin in brain parenchyma increased with aging, especially in senescence accelerated mice (SAMP8), which showed age-related deficits in learning and memory. In this study, in order to examine morphological events related to the age-related increase of the brain accumulation of serum albumin, the transvascular passage of blood-borne horseradish peroxidase (HRP) and ultrastructural features of microvessels were examined in the hippocampus, cerebellum and pons of SAMP8 and SAMR1 (control) mice. Ultrastructural examination of the hippocampus showed that the staining for HRP was occasionally spreading throughout the parajunctional cytoplasm of the endothelial cell of aged SAMP8 mice, but not in young SAMP8 mice nor in SAMR1 mice. The number of vessels showing the staining reaction for HRP in the parajunctional cytoplasm of the endothelial cells in aged SAMP8 mice increased significantly compared with that in the others. Electron microscopic morphometry showed that there were no significant differences among the number of HRP-positive vesicles per unit area of the endothelial cell cytoplasm in young and old mice of both strains. The staining reaction for HRP was not seen in the basal lamina of microvessels and the perivascular neuropil in all mice examined. Perivascular lipofuscin-like granules and collagen deposits, swelling of astroglial perivascular endfeet and perivascular cells containing foamy, lipid-like droplets were frequently found in several brain regions of aged SAMP8 mice. The perivascular cells with a few lipid like droplets and more electron-homogeneous lysosomes were occasionally seen in SAMR1 and young SAMP8, while the other findings were scarcely observed in SAMR1 and young SAMP8 mice. These findings suggest that the blood-brain barrier to HRP was preserved in microvessels in three brain regions of SAM mice but the blood microvessels showed some age-related ultrastructural alterations in SAMP8 brains. Uncontrolled passage of HRP through the parajunctional cytoplasm of the endothelial cells may partly contribute to the age-related increase of accumulation of serum albumin in SAMP8 brains. PMID- 11378255 TI - Deterioration in learning and memory of fear conditioning in response to context in aged SAMP8 mice. AB - This study examined age-dependent deficits in the learning and memory of fear conditioning, using a newly developed senescence-accelerated mouse (SAMP8) model of age-related brain dysfunction and its genetically related inbred strain (SAMR1). The mice were classically conditioned to tone by giving aversive foot shocks in a distinct experimental box (context). After conditioning, fear in response to the original context without the tone and to the tone in a different context were tested with no shocks. Freezing behavior was used as a reliable index of fear. At 4 and 8 months, contextual fear was weaker in the accelerated senescence-prone SAMP8 mice than in the accelerated senescence-resistant SAMR1 mice. However, at 1 and 2 months, both SAMP8 and SAMR1 mice showed significant contextual fear to equivalent levels. Aging did not affect the fear response to tone. These results indicate that SAMP8 mice have age-related learning and memory deficits in their fear response evoked by contextual but not explicit tone stimuli. Age-related hippocampal dysfunction is suggested to be the cause of these age-related deficits in contextual fear conditioning in SAMP8 mice. PMID- 11378256 TI - The acetylcholine release enhancer linopirdine induces Fos in neocortex of aged rats. AB - Centrally acting cholinergic agents induce the immediate early gene c-fos in the rat brain resulting in transient increases of Fos protein, most notably in the cerebral cortex. In this study we have monitored by Fos immunohistochemistry the effect of the acetylcholine release enhancer linopirdine (DUP996) on the immediate early gene c-fos in brains of 3 months and 30 months old rats. In young rats linopirdine had only a marginal effect on Fos expression. In contrast, in aged rats linopirdine caused widespread expression of Fos throughout neocortex. In somatosensory cortex, the induction of the c-fos gene by linopirdine was nearly completely blocked by atropine and scopolamine and strongly attenuated by the NMDA receptor blockers CPP and MK-801. The results suggest that the age related decline in acetylcholine release in rodents can be partially compensated for by administration of linopirdine. PMID- 11378257 TI - Nomifensine reveals age-related changes in K(+)-evoked striatal DA overflow in F344 rats. AB - To investigate the influence of age-associated changes in DA uptake on measures of potassium-stimulated DA overflow in the striatum, microdialysis was conducted in anesthetized young (6-month-old) versus aged (24-month-old) F344 rats. Extracellular levels of DA, DOPAC, and HVA were measured under basal and potassium-stimulated (10, 25, 50, & 100 mM) conditions. Basal levels of DA and metabolites did not differ significantly between the two age groups. At the 50 and 100 mM concentrations, potassium stimuli significantly increased DA overflow and decreased DOPAC and HVA--effects that did not differ with age. The addition of the DA uptake inhibitor nomifensine (100 microM) to the perfusion solutions revealed differences between the two age groups. Nomifensine augmented potassium evoked DA overflow at the 50 mM concentration in both groups, but only amplified the effect of the 100 mM concentration in the young animals. The results demonstrate that decreased DA transporter function in aged rats masks age-related differences in K(+)-evoked striatal DA release when microdialysis methods are used, resulting in net equalization of K(+)-evoked striatal DA overflow in young versus aged F344 rats. PMID- 11378258 TI - cAMP and excitability in neuroendocrine cells during reproductive senescence. AB - Excitability changes during reproductive senescence were investigated in the neurosecretory caudodorsal cells (CDCs) that control egg laying in the mollusc Lymnaea stagnalis. CDCs in the isolated central nervous system (CNS) were exposed to different discharge inducing treatments. Senescent CDCs (of animals 8 weeks after laying their last egg mass) and inhibited (I-) state CDCs (of egg-laying (EL) animals) were used. We showed that senescent and I-state CDCs closely resemble each other electrophysiologically. Electrical stimulation did not induce an afterdischarge in either type of CDC but exposure to release products of CDCs from EL animals or to saline with high potassium concentration did induce discharge activity. Also, 8-chlorophenylthio (8-CPT)-cAMP (10(-5) M) induced discharge activity. Exposure to the phosphodiesterase inhibitor 3-isobutyl-1 methylxanthine (IBMX) (10(-3) M) or to the adenylate cyclase activator Forskolin (10(-4) M), restored afterdischarge induction by electrical stimulation. Application of IBMX (10(-3) M) and Forskolin (10(-4) M) together induced discharges in the absence of electrical stimulation. Our results suggest that in senescent CDCs changes in the intracellular cAMP pathway may underlie afterdischarge failure. PMID- 11378259 TI - Meeting summary of the Tenth International Symposium on autologous blood and marrow transplantation. PMID- 11378260 TI - Treatment of autoimmune diseases by hematopoietic stem cell transplantation. AB - Remarkable advances have been made in bone marrow transplantation (BMT), which now has become a powerful strategy for the treatment of leukemia, aplastic anemia, congenital immunodeficiency disorders, and autoimmune diseases. Using various animal models, allogeneic BMT has been found to be useful in the treatment of autoimmune diseases. In MRL/lpr mice, which are radiosensitive (<8.5 Gy) and are an animal model for autoimmune disorders, conventional BMT resulted in only transient effects; the manifestations of the autoimmune diseases recurred 3 months after BMT. However, the combination of BMT plus bone grafts (to recruit donor stromal cells) was capable of preventing the recurrence of autoimmune diseases in MRL/lpr mice. This strategy was found to be ineffective in the treatment of MRL/lpr mice that had developed autoimmune diseases, because these mice were more sensitive to the effects of radiation after the onset of lupus nephritis due to uremic enterocolitis. We have recently discovered a safer strategy for treatment of autoimmune diseases, which includes fractionated irradiation (5.5 Gy x 2) (day -1) followed by portal venous injection (day 0) plus intravenous injection (day 5) of donor unfractionated bone marrow cells. We successfully treated autoimmune diseases in MRL/lpr mice using this strategy; 100% of MRL/lpr mice treated in this fashion survive >1 year after treatment. We identified the mechanisms underlying the components of this approach and have found that stromal cells play a crucial role in successful BMT. In this review, the conditions essential for successful allogeneic BMT are discussed. PMID- 11378261 TI - Clonal hemopoiesis and risk of thrombosis in young female patients with essential thrombocythemia. AB - OBJECTIVE: Several studies demonstrated a high prevalence of nonrandom X chromosome inactivation pattern (X-CIP) in essential thrombocythemia (ET). This study explored the incidence of clonal hemopoiesis in myeloid precursors and endogenous erythroid colonies (EECs) in ET patients and its correlation with thrombotic manifestations. MATERIALS AND METHODS: Clonal analysis of hemopoiesis using X-CIP was performed in 40 female patients with ET. Median age was 40.5 years (range 20-64), and median platelet count at testing time was 700 x 10(9)/L (range 220-1300 x 10(9)/L). Patients older than 65 years were excluded to reduce age-related skewing. Clonality was assessed on neutrophils, platelets, EECs, and bone marrow CD34(+) cells. RESULTS: Eight (20%) of 40 patients developed thrombosis mainly at diagnosis. Clonal hemopoiesis was found in 17 (42.5%) patients, 15 (37.5%) had polyclonal hemopoiesis, and 8 (20%) were considered uninterpretable due to constitutive skewing. Clonality was confirmed on purified CD34(+) subpopulations from bone marrow, documenting that clonality does not appear lineage-restricted. There were no statistical differences in age at diagnosis, median platelet count at testing time, and length of follow-up. Thrombotic episodes were significantly more frequent in the monoclonal group (p = 0.04, Fisher exact test). CONCLUSIONS: Young female patients with ET exhibiting a clonal pattern of hemopoiesis by X-CIP analysis are at higher risk for thrombosis. X-CIP analysis may contribute to defining the individual risk leading to appropriate treatment. X-CIP will allow a correct diagnosis in patients with latent myeloproliferative disorders and thrombosis in unusual sites. Clonal hemopoiesis is easily recognized by X-CIP, but its applicability is limited to the female sex and is hampered by the presence of age-related or constitutive skewing. PMID- 11378262 TI - Serum soluble transferrin receptor concentration is an accurate estimate of the mass of tissue receptors. AB - OBJECTIVE: Serum levels of the soluble transferrin receptor (sTfR) vary depending on the erythropoietic activity and iron status. In vitro, sTfR shed in the incubation medium correlates well with cellular TfR, but this relationship has never been established in vivo. To determine the value of serum sTfR as a quantitative marker of the body mass of tissue TfR, we designed experiments to examine the correlation between serum sTfR and tissue TfR in rats with various degrees of erythropoietic activity or iron status. MATERIALS AND METHODS: We studied changes in erythropoietic activity in normal rats as well as in animals experiencing hemolysis, phlebotomy-induced iron deficiency, transfusion- or thiamphenicol-induced erythroid aplasia, or inflammation. At the end of follow up, ferrokinetic studies were performed and animals were sacrificed. Organs were isolated and homogenized to determine the total mass of tissue TfR from the sum of tissue solubilized TfR in the bone marrow, spleen, liver, and blood cells (direct method). An indirect method was developed to derive the corporeal mass of tissue TfR from a representative marrow sample. RESULTS: As expected, serum sTfR and total mass of tissue TfR varied as a function of iron status and erythropoiesis. Relative erythroid expansion in the spleen was greater than in the bone marrow. With the exception of phlebotomized animals, the indirect method correlated very well with direct measurements of the total mass of tissue TfR (r = 0.97, p < 0.0001). There was a close relationship between the total mass of tissue TfR and the total mass of serum sTfR (r = 0.79, p < 0.0001). Serum sTfR represented approximately 5-6% of the total mass of tissue TfR in most experimental situations, but this ratio was twice as high during iron-restricted erythropoiesis. In addition, the ratio could be higher or lower in nonsteady state situations, because changes in tissue TfR occurred faster than those of serum sTfR. CONCLUSIONS: Serum sTfR represents a constant proportion of the total mass of tissue TfR over a wide range of erythropoietic activity. However, iron deficiency results in a higher proportion of serum sTfR, and the pace of change in serum sTfR levels is slower than that of tissue TfR mass. PMID- 11378263 TI - Isolation and characterization of canine hematopoietic progenitor cells. AB - The purpose of this study was to purify and characterize canine hematopoietic progenitor cells for surface antigen phenotype and reconstitution ability. Canine hematopoietic progenitor cells were isolated by density gradient sedimentation, lineage depletion with monoclonal antibodies, and fluorescence-activated cell sorting (FACS) for selection of cells with low-forward and right-angle scatter that were rhodamine 123 (Rh-123)(dull). Isolated cells were characterized for expression of CD34, c-kit, and Flt-3. A canine/murine xenograft model and a mixed chimerism assay were used to examine the in vivo proliferative potential of isolated cells. The lineage-positive (Lin(+)) cells represented 80 +/- 11% (n = 22) of the input mononuclear cells. Lineage depletion resulted in a fourfold increase in colony-forming unit granulocyte/monocyte (CFU-GM), a 2.5-fold increase in burst-forming unit-erythroid (BFU-E), and a twofold increase in the number of Rh-123(dull) cells over nonlineage-depleted bone marrow mononuclear cells (BMMCs). Lineage depletion led to a 2.7-fold enrichment of CD34 cells, a 10.4-fold enrichment of c-kit cells, and a 10.8-fold enrichment of CD34/c-kit(+1) cells over total BMMCs. Nineteen percent of lineage-negative (Lin(-)) cells were positive for Flt-3. Injection of canine cells into irradiated (400 rads) NOD/SCID mice resulted in the detection of canine CD45(+) cells with BMMCs, Lin(-) cells, or Rh-123(dull) cells. Transplantation of purified Lin(-) cells in dog leukocyte antigen-matched littermates resulted in low-level engraftment for at least 10 weeks. The development of methods for purification and characterization of canine hematopoietic progenitor cells should enhance the utilization of the canine model for a variety of experimental and therapeutic purposes. PMID- 11378264 TI - STAT3 is constitutively active in some patients with Polycythemia rubra vera. AB - OBJECTIVE: Polycythemia vera is a clonal stem cell disorder characterized by hyperproliferation of the erythroid, myeloid, and megakaryocytic lineages. While it has been shown that progenitor cells of P. vera patients are hypersensitive to several growth factors including erythropoietin, insulin-like growth factor-1, thrombopoietin, interleukin-3, and granulocyte/monocyte colony-stimulating factor, the molecular pathogenesis of this disease remains unknown. Growth factor hypersensitivity could be mediated by changes in signal transduction pathways. We therefore investigated a common downstream effector of cytokines, the signal transducers and activators of transcription (STATs). A constitutive activation of STAT factors could explain the increased proliferation of P. vera cells even in the absence of growth factor stimulation. METHODS: Peripheral granulocytes from patients with P. vera and from healthy volunteers were assayed for STAT1, 3, and 5 DNA binding by electrophoretic mobility shift assay. RESULTS: Four of 14 P. vera patients analyzed showed constitutive STAT3 DNA binding in unstimulated peripheral granulocytes, while none of the 17 healthy volunteers tested did. None of the subjects showed constitutive STAT1 or STAT5 activity. Western blotting demonstrated that, in the three patients, STAT3 is constitutively phosphorylated on Tyr 705, whereas it is unphosphorylated in the other patients and in controls. Interestingly, constitutive STAT3 activity did not correlate with the duration of disease or the treatment regimen. It was observed in a recently diagnosed patient and in two patients treated only with phlebotomy. CONCLUSION: Our data suggest that constitutive phosphorylation and activation of STAT3 is not a secondary event induced by mutagenizing agents or by prolonged hyperproliferation of hematopoietic cells, but rather represents a primary molecular aberration. Constitutively active STAT3 may contribute to the growth factor hypersensitivity of P. vera cells. PMID- 11378265 TI - UCN-01 induces cytotoxicity toward human CLL cells through a p53-independent mechanism. AB - OBJECTIVES: UCN-01, a novel protein kinase C inhibitor, is currently being tested in phase I clinical trials after being noted to induce apoptosis in lymphoid cell lines. We sought to study the in vitro activity of UCN-01 against human chronic lymphocytic leukemia (CLL) cells and potential mechanisms of action for inducing this cytotoxicity. METHODS: Detailed in vitro studies were performed from tumor cells derived from patients with CLL cells following attainment of written informed consent. RESULTS: The 50% loss of viability (LC(50)) in mononuclear cells from CLL patients (n = 10) following exposure to UCN-01 for 4 days was 0.4 microM (95% CI +/- 0.21; range 0.09-1.16). Loss of viability in human CLL cells correlated with early induction of apoptosis. Exposure of CLL cells to 0.4 and 5.0 microM of UCN-01 resulted in decreased expression of p53 protein. We therefore investigated the dependence of UCN-01 on intact p53 by exposing splenocytes from wild-type (p53(+/+)) and p53 null (p53(-/-)) mice, which demonstrated no preferential cytotoxicity when compared to the marked differential induced by F-Ara-A and radiation. CONCLUSIONS: UCN-01 has significant in vitro activity against human CLL cells that appears to occur independent of p53 status. While demonstration of in vitro cytotoxicity does not establish in vivo efficacy, the findings described support the early introduction of UCN-01 into clinical trials for patients with B-CLL. PMID- 11378266 TI - Leukemia blast-induced T-cell anergy demonstrated by leukemia-derived dendritic cells in acute myelogenous leukemia. AB - OBJECTIVE: To elucidate the mechanism of immunologic escape of leukemia cells and establish an effective anti-leukemia immunotherapy, we attempted to generate dendritic cells from leukemia cells in patients with acute myelogenous leukemia (AML). Using these leukemia-derived dendritic cells, we investigated leukemia cell-associated T-cell anergy. MATERIALS AND METHODS: Leukemia cells of 30 patients with AML were cultured with granulocyte-macrophage colony-stimulating factor, interleukin-4, and tumor necrosis factor-alpha. Cultured leukemia cells were evaluated for antigen-presenting ability by mixed leukocyte culture (MLC). Normal lymphocytes, which were cocultured with leukemia blasts in the first MLC, were cultured with leukemia-derived dendritic cells in the second MLC. RESULTS: In cultures of leukemia cells from 21 of 30 patients examined, cells with stellate morphology and cell fractions with CD1a(+) and/or CD83(+) were present. Autologous MLC using lymphocytes obtained in remission phase as responders as well as allogeneic MLC demonstrated antigen-presenting ability in leukemia derived dendritic cells. Leukemia cells of FAB-M0, M1, M2, M3, or M6 morphology/phenotype gave rise to dendritic cells as well as leukemia cells of M5. The leukemic origin of dendritic cells was suggested by in situ hybridization. By coculture with CD80(-) leukemia blasts, the response of normal lymphocytes to leukemia-derived dendritic cells cultured from the same individual as that of leukemia blasts was markedly reduced, compared with the lymphocytes cultured with leukemia blasts from a different individual as leukemia blasts. CONCLUSIONS: Escape of leukemia cells from anti-leukemia immunity may be associated with T-cell anergy caused by leukemia blasts. The results of the present study suggest that leukemia-derived dendritic cells can be applied efficiently in anti-leukemia immunotherapy. PMID- 11378267 TI - Injection of CD4(+) and CD8(+) cells with donor or host accessory cells induces acute graft-vs-host disease in human skin in immunodeficient mice. AB - OBJECTIVE: We examined cell subsets with respect to cutaneous graft-vs-host disease by cell sorting selection of subsets of human mononuclear cells and injecting the subsets subcutaneously in a mouse model. MATERIALS AND METHODS: Cell suspensions containing cultured human epidermal cells and dermal fibroblasts from a single donor mixed with lymphoid cell subsets positively selected using the FACSVantage cell sorting instrument and/or MACS cell isolation kits from unrelated individuals were injected into immunodeficient mice. This model is known to generate human skin with histologic findings similar to human graft-vs host disease. RESULTS: Donor T-cell subsets CD4(+) and CD8(+) plus either host or donor CD14(+) cells were necessary to cause acute cutaneous graft-vs-host disease. Although graft-vs-host disease can result from recognition of class I antigens expressed on human cutaneous cells by donor peripheral blood mononuclear cells, additional recognition of class II antigens expressed on host mononuclear cells resulted in more severe histologic manifestations. Dendritic cells that differentiated from donor and host monocytes also showed competent accessory cell function in this system. CONCLUSIONS: Based on this model, human cutaneous graft vs-host disease was caused by donor CD4(+) cells and CD8(+) cells activated through recognition of host antigens, including class I and class II antigens presented by either donor or host CD14(+) cells or dendritic cells. PMID- 11378268 TI - Antiapoptotic protein Bcl-x(L) is up-regulated during megakaryocytic differentiation of CD34(+) progenitors but is absent from senescent megakaryocytes. AB - OBJECTIVE: The expression of Bcl-x(L) has been shown to be regulated during the maturation process of different hematopoietic cell lineages (i.e., erythroid cells, neutrophils, monocytes/macrophages). In the present study, we examined the expression of Bcl-x(L) in megakaryocytes derived from CD34(+) progenitors and in the megakaryoblastic cell line UT7. MATERIALS AND METHODS: Expression of Bcl-x(L) was analyzed in CD41(+) cells cultured in the presence of thrombopoietin and in UT7 cells treated with phorbol diester by Western blot, flow cytometry, and immunocytochemistry analysis. Apoptosis was determined at different culture times by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling and propidium iodide uptake. RESULTS: Bcl-x(L) but not Bcl-2 was up-regulated in the megakaryocytic population (CD41(+)) during the first 15 days of culture, which was consistent with the pattern of Bcl-x(L) expression in UT7 cells differentiated to megakaryocytes by incubation with phorbol diester. However, by day 20 of culture, the levels of Bcl-x(L) in CD41(+) cells were greatly reduced, and this expression pattern was accompanied by an increase in the number of apoptotic cells. At this culture time, we detected the presence of cytoplasmic fragments resembling proplatelets with prominent Bcl-x immunostaining, most likely due to the Bcl-x(L) isoform, in close proximity to Bcl-x(-) senescent megakaryocytes. The presence of Bcl-x(L) but not of Bcl-2 in platelets was confirmed by Western blot analysis. CONCLUSION: Although little is known regarding the functional significance of survival proteins within the megakaryocytic compartment, the changes in the Bcl-x(L) expression pattern observed in UT7 and CD41(+) cells may play a role in the survival of developing megakaryocytes and the lifespan of mature platelets. PMID- 11378269 TI - Distinct hematopoietic support by two human stromal cell lines. AB - OBJECTIVE: The hematopoietic microenvironment is complex, and the role of myofibroblast in its function is crucial. In order to obtain a stable model reflecting this particular cell type, we have previously established human bone marrow cell lines from primary myofibroblastic Stro1(+) population (pStro1(+)). We placed HPV16 E6 and E7 expression under the control of different promoters. Here, we have characterized and studied the hematopoietic support for two cell lines corresponding to the promoters alpha-SM (alphaSM-56 line) and SV40 (SV40-56 line). MATERIALS AND METHODS: The expression profile was analyzed at the RNA level by gene array and at the protein level by Western blot, flow cytometry, and ELISA. Hematopoietic support determined using colony-forming unit (CFU) and stroma-adherent colony-forming cell (SA-CFC) assays. RESULTS: The phenotype of cell lines was not significantly modified compared with primary myofibroblastic cells. They secreted a broad spectrum of hematopoietic cytokines and nonspecific mediators. The two lines allowed the growth of hematopoietic precursors and had different support capabilities. CONCLUSIONS: We have extensively characterized two novel human bone marrow stromal cell lines. They retained a myofibroblastic phenotype and have substantial but different hematopoietic support capabilities. These lines provided a basis for determining stromal factors involved in stem cell regulation. PMID- 11378270 TI - Suppression of apoptosis and granulocyte colony-stimulating factor-induced differentiation by an oncogenic form of Cbl. AB - OBJECTIVE: The retroviral oncogene v-Cbl causes pre-B cell lymphomas and myeloid leukemias in mice, and its Drosophila homologue is oncogenic, causing enhanced receptor tyrosine kinase signaling. The human Cbl gene resides at 11q23. The aim of this study is to determine the effect of oncogenic Cbl on growth-regulating responses. MATERIALS AND METHODS: The oncogenic mutant of Cbl (CblDelta1-357) was transfected into factor-dependent 32Dcl3 myeloid cells. Consequently, cell survival and differentiation were measured. Lyn, Syk, MAP kinase, and phosphatidylinositol 3'(PI3')-kinase activities, protein phosphorylation, Bcl-2 promoter activity, ubiquitination, and levels of Bcl-2, Bax, Bad, and Bcl-x(L) were determined. In addition, the effect of v-Cbl on TF-1 cell survival upon granulocyte-macrophage colony-stimulating factor withdrawal was studied. RESULTS: 32Dcl3 and TF-1 cells expressing v-Cbl showed resistance to apoptosis upon growth factor withdrawal, and 32Dcl3 cells completely failed to respond to granulocyte colony-stimulating factor's induction of differentiation. Basal activities of Lyn, Syk, and PI3'-kinase were elevated in the v-Cbl line. There was neither enhanced tyrosine phosphorylation of cellular protein content, Cbl, or Jak2, nor serine phosphorylation of MAP kinase or Akt. After factor withdrawal, the level of Bcl-2 was greater in v-Cbl cells than in control cells. CONCLUSIONS: Neither increased Bcl-2 promoter activity nor decreased ubiquitination of Bcl-2 could account for increased Bcl-2 levels. v-Cbl-expressing 32Dcl3 cells were resistant to differentiation. v-Cbl suppresses apoptosis and differentiation, possibly through enhancement of Lyn, Syk, and PI3'-kinase activities and Bcl-2. PMID- 11378271 TI - Functional identification of secondary mutations inducing autonomous growth in synergy with a truncated interleukin-3 receptor: implications for multi-step oncogenesis. AB - OBJECTIVE: A truncated common beta chain (Deltabeta(C)) of the interleukin-3 (IL 3) receptor complex was previously identified as a key factor in inducing autonomous growth of IL-3-independent mutants. Expression of Deltabeta(C) in IL-3 dependent hematopoietic cells does not result in immediate factor-independent growth, but increases the frequency of obtaining autonomous mutants by three to four orders of magnitude. This study was designed to delineate the mechanisms by which Deltabeta(C) increases the frequency to autonomous growth. DESIGN AND METHODS: Retroviral vectors were used to express Deltabeta(C) into IL-3-dependent myeloid cells, which were then tested for factor-independent growth. To determine if secondary genetic events were required for conversion to autonomous growth, elements of the Cre-loxP recombinant system were used to excise Deltabeta(C) in factor-independent clones. RESULTS: Excision of Deltabeta(C) in factor independent clones revealed two types of phenotypes: reversion to factor dependent growth (1/8) or continued IL-3-dependent growth (7/8). Analysis of cells that remained factor independent revealed constitutive activation of STAT5, not observed in factor-dependent revertants. Analysis of revertant cells demonstrated the presence of interacting secondary mutations that synergize with Deltabeta(C)-induced proliferation. A cysteine residue within the truncated extracellular domain of Deltabeta(C) was also found to be required for its oncogenic potential, supporting a model of dimerization for receptor activation. CONCLUSIONS: The high incidence of obtaining factor-independent mutants from cells expressing Deltabeta(C) results from the selection of mutations that either complement Deltabeta(C) expression to promote proliferation or that singly or in synergy with other secondary mutations negate the requirement of Deltabeta(C) expression for proliferation. PMID- 11378272 TI - Tissue source dictates lineage outcome of human fetal CD34(+)CD38(-) cells. AB - OBJECTIVE: The translocation from fetal liver hematopoiesis to secondary organs occurs during the second trimester of human gestation. It has been hypothesized that stem cells migrate and acquire lineage potential based on cues specific to the adopted microenvironment. We evaluated primitive hematopoietic cell populations in the fetal human to determine if lineage restriction precedes or follows translocation to sites of hematopoietic activity including thymus, spleen, bone marrow, and liver. METHODS: Sets of hematopoietic tissues from individual second-trimester human abortuses were used to compare and quantitate the lineage outcome of immunophenotypically primitive cells from each of the hematopoietic organs using ex vivo myeloid and lymphoid differentiation systems. RESULTS: Despite uniformity in immunophenotype, functional capabilities were highly restricted by the tissue of origin and alteration in the ex vivo differentiation context did not lead to a change in differentiation outcome. CONCLUSION: Translocation of primitive cells from fetal liver to tissues of mature hematopoietic activity is associated with tissue-specific, quantitative changes in differentiation potential that are unresponsive to alternative differentiation environments. These data suggest that multipotentiality is lost prior to or upon stem-cell migration in the developing human. It is not persistent with residence in a secondary hematopoietic organ. PMID- 11378273 TI - Circulating factors may be responsible for murine strain-specific responses to mobilizing cytokines. AB - OBJECTIVE: To determine if circulating factors influence strain-specific responses to administration of hematopoietic stem-cell mobilizing cytokines, a murine model was employed. METHODS: Plasma aliquots from intact DBA2, Balb/c, and C57Bl/6 mice were injected into intact Balb/c mice prior to delivery of mobilizing cytokines. Control Balb/c mice were injected with mobilizing cytokines alone. Plasma from hemi-body irradiated Balb/c mice, known to inhibit mobilization, was also injected into Balb/c mice. Twenty-four hours later, spleen cells were harvested and assayed for granulocyte-macrophage colony-forming cells (GM-CFC) and high-proliferative-potential colony-forming cells (HPP-CFC). Simultaneously harvested blood aliquots were assayed for CD45(+)/CD34(+) cells using flow cytometric techniques. RESULTS: Mice receiving plasma from any source demonstrated significant inhibition of mobilization of HPP-CFC and GM-CFC to the spleen as compared to mobilized controls; for HPP-CFC, plasma from C57Bl/6 mice was more inhibitory than plasma from Balb/c (p = 0.001) or from DBA2 mice (p = 0.01), while for GM-CFC, plasma from C57Bl/6 mice was more inhibitory than Balb/c plasma but not more inhibitory than DBA2 plasma. Mice injected with plasma from previously irradiated Balb/c mice exhibited the expected HPP-CFC and GM-CFC mobilization inhibition, which was not statistically different from the inhibition seen in animals that received C57Bl/6 plasma. Mobilization of CD34(+)/CD45(+) cells to the blood also appeared to be inhibited by pretreatment with C57Bl/6 plasma, but not DBA2 plasma. CONCLUSION: These data suggest that strain-specific patterns of mobilization may be influenced by a circulating mobilization inhibitor(s). PMID- 11378274 TI - The effects of SCF/G-CSF prestimulation on radiation sensitivity and engraftment in nonmyeloablated murine hosts. AB - OBJECTIVE: Previous studies have shown improved engraftment in a murine model when granulocyte colony-stimulating factor (G-CSF) and stem cell factor (SCF) were administered for 5 days prior to irradiation, with significant levels of engraftment in the growth factor-preconditioned group even at very low radiation doses. We sought to explore the mechanisms behind this effect. METHODS: The radiation sensitivity of mice with or without 5 days of prestimulation with G-CSF (200 microg/kg/d) and SCF (50 microg/kg/d) was compared. To further evaluate whether growth factor prestimulation enhances engraftment by mobilization of hematopoietic progenitors into peripheral blood, thus creating less endogenous competition within the marrow compartment, female mice were pretreated with 5 days of G-CSF/SCF or control diluent. Engraftment of 40 x 10(6) peripheral blood stem cells (PBSCs) harvested from G-CSF/SCF-mobilized male mice was compared in the two recipient groups. RESULTS: There was no difference in survival between the pretreated and control mice at the radiation doses tested. Additionally, there was no significant difference in the recovery of blood counts, bone marrow cellularity, colony-forming unit (CFU) content, or stem cell numbers assessed 4 months later in a competitive repopulation model. Engraftment levels of male cells did not differ between G-CSF/SCF-pretreated and control recipients, and could be detected in 30% of recipients at 20-24 weeks (4/12 in each group) at overall levels of 0.1-1%. CONCLUSIONS: The enhanced engraftment in cytokine pretreated recipients is unlikely to be due to increased endogenous stem-cell killing or to the creation of endogenous marrow "space" by egress of endogenous stem cells after cytokine prestimulation. PMID- 11378275 TI - Lifelong hematopoiesis in both reconstituted and sublethally irradiated mice is provided by multiple sequentially recruited stem cells. AB - OBJECTIVE: To evaluate the dynamics of stem cell production to hematopoiesis, the number of active stem cell clones and the lifespan of individual clones were studied. MATERIALS AND METHODS: The clonal contribution of primitive hematopoietic stem cells (HSC) responsible for long-term hematopoiesis was determined using two approaches. In one model, irradiated female mice were reconstituted with retrovirally marked male hematopoietic cells. In the second model, mice were irradiated sublethally without hematopoietic cell transplantation. In both models, bone marrow cells were serially sampled from the same mouse throughout a 12- to 20-month period and injected into irradiated recipients for analysis of day 10 colony-forming unit-spleen (CFU-S). The donor origin of CFU-S was determined by the presence of retrovirally marked cells or cells with chromosomal aberrations. RESULTS: The results of the two essentially different models show that 1) hematopoiesis is mainly the product of small clones of hematopoietic cells; 2) the lifespan of the majority of clones is only 1 to 2 months; 3) the clones usually function locally; and 4) the vast majority of the clones replace one another sequentially. Primitive HSCs capable of producing long lived clones (about 10% among all clones), which exist during the entire life of a mouse, were detected by the radiation-marker technique only. CONCLUSION: Multiple short-living clones (at least on the level of CFU-S production) comprise the vast majority of the active stem cells in transplanted recipients or after endogenous recovery from sublethal irradiation. PMID- 11378277 TI - Plants used in home medicine in the Zenta River basin, Northwest Argentina. AB - The use of herbal pharmacopoeia in the domestic life of a rural community in the west of the Oran Department, Salta Province, Argentina is reported in this article. One hundred and seventeen plant species belonging to 52 families and 98 different genera were included. Their vernacular name, popular use, administration, used parts and ways of obtaining are given. It is concluded that home medicine constitutes a relevant resource in the area. PMID- 11378276 TI - Antiulcer activity of Opuntia ficus indica (L.) Mill. (Cactaceae): ultrastructural study. AB - In Sicily folk medicine, Opuntia ficus indica (L.) Mill. cladodes are used for the treatment of gastric ulcer. We studied the effect of administration of lyophilized cladodes on experimental ethanol-induced ulcer in rat. In this paper, we report the ultrastructural observations of gastric mucosa. The ultrastructural changes were observed by trasmission electronic microscopy (TEM) confirming the protective effect exercised by administration of lyophilized cladodes. Pre treatment test in rats revealed a protective action against ethanol-induced ulcer. Probably, the mucilage of Opuntia ficus indica is involved. PMID- 11378278 TI - Hypoglycemic effect of Suaeda fruticosa in streptozotocin-induced diabetic rats. AB - The purpose of this study was to examine the hypoglycemic activity of the aqueous extract of the aerial part of Suaeda fruticosa (SF) in normal and streptozotocin induced diabetic rats. The aqueous extract was administered intravenously (i.v.) and the blood glucose changes were determined within 4 h after starting the treatment. Plasma insulin, cholesterol and triglycerides levels were also determined. The aqueous extract at a dose of 192 mg/kg produced a significant decrease in blood glucose levels in normal rats (P < 0.05), and even more in diabetic rats (P < 0.001). This hypoglycemic effect might be due to an extra pancreatic action of the aqueous extract of SF, since that the levels of plasma insulin were unchanged between the values before and after treatment. In the other hand, the effect of the aqueous extract on the plasma cholesterol were also significant in both normal and diabetic rats (P < 0.05). But, there is no significant effect of SF on plasma triglycerides in both groups. In order to characterize the active principle(s), which could be responsible for the therapeutic effect, preliminary phytochemical analysis of the aqueous extract of the plant has been investigated. PMID- 11378279 TI - Screening of antibiotic resistant inhibitors from local plant materials against two different strains of Pseudomonas aeruginosa. AB - The methanolic extracts of 19 Jordanian plants were combined with seven different antibiotics and applied to check the inhibitory effects of the combination on the resistance of Pseudomonas aeruginosa. A resistant strain of Ps. aeruginosa, which was isolated from a patient and a standard strain of the same microorganism were used in this study. Our results showed that there are significant variations in the effects of some combinations used on the resistant and the standard strains probably due to structural changes. Almost all the plant materials used in combination with penicillin G and erythromycin allowed full growth of the standard strain, while the combination with some plant materials like Gundelia tournefortii L. and Lepidium sativum L. inhibited the growth of the resistant strain. Chloramphenicol, gentamicin and cephalosporin can be given advantageously with almost all the plant materials used with few exceptions on the resistant strain. Nalidixic acid activity was improved significantly when combined with all plant materials and tested on standard strain. On the other hand, its activity on the resistant strain was slightly improved using the same combinations. PMID- 11378280 TI - The anti-inflammatory, analgesic and antipyretic activity of Nigella sativa. AB - The aqueous extract of Nigella sativa (N. sativa) was investigated for anti inflammatory, analgesic and antipyretic activities in animal models. The extract has an anti-inflammatory effect demonstrated by its inhibitory effects on Carrageenan induced paw edema. It also produced significant increase in the hot plate reaction time in mice indicating analgesic effect. However, N. sativa crude suspension had no effect on yeast induced pyrexia. This study therefore, supports its use in folk medicine both as analgesic and anti-inflammatory agent and calls for further investigations to elucidate its mechanism of action. PMID- 11378281 TI - Hypericum perforatum as a nootropic drug: enhancement of retrieval memory of a passive avoidance conditioning paradigm in mice. AB - Depression, among other non-cognitive symptoms, is common in patients with dementia. The effect of Hypericum perforatum (St. John's Wort) extract, with well documented antidepressant activity, was tested on memory retrieval 24 h after training on a one-trial passive avoidance task in mice. Acute administration of Hypericum extract (4.0, 8.0, 12.0, and 25.0 mg/kg i.p.) before retrieval testing increased the step-down latency during the test session. The same doses of Hypericum extract, on the other hand, failed to reverse scopolamine-induced amnesia of a two-trial passive avoidance task. The involvement of serotonergic, adrenergic, and dopaminergic mechanisms in the facilitatory effect of Hypericum extract on retrieval memory was investigated. Pretreatment of the animals with serotonergic 5-HT1A receptor antagonist (-)-pindolol (0.3, 1.0, and 3.0 mg/kg), serotonergic 5-HT2A receptor blocker spiperone (0.01, 0.03, and 0.1 mg/kg), alpha adrenoceptor antagonist phentolamine (1, 5, and 10 mg/kg), beta receptor antagonist propranolol (5, 7.5, and 10 mg/kg), dopaminergic D1 receptor antagonist SCH 23390 (0.01, 0.05, and 0.1 mg/kg), and dopaminergic D2 receptor antagonist sulpiride (5, 7.5, and 10 mg/kg) revealed the involvement of adrenergic and serotonergic 5-HT1A receptors in the facilitatory effect of Hypericum extract on retrieval memory. It is concluded that Hypericum extract may be a better alternative for treatment of depression commonly associated with dementia than other antidepressants known to have anticholinergic side effects causing delirium, sedation and even exacerbating already existing impaired cognition. In dementias of old age, Hypericum perforatum would, therefore, serve as one medication targeting both depression and amnesia with lower potential side effects. PMID- 11378282 TI - Inhibitory effects of methanol extract of Cyperus rotundus rhizomes on nitric oxide and superoxide productions by murine macrophage cell line, RAW 264.7 cells. AB - The rhizomes of Cyperus rotundus (C. rotundus) have been used in oriental traditional medicines for the treatment of stomach and bowel disorders, and inflammatory diseases. Nitric oxide (NO) and superoxide (O2-) are important mediators in the pathogenesis of inflammatory diseases. This study was undertaken to address whether the metanol (MeOH) extract of rhizomes of C. rotundus could modulate NO and O2- productions by murine macrophage cell line, RAW 264.7 cells. The MeOH extract of rhizomes of C. rotundus showed the inhibition of NO production in a dose-dependent manner by RAW 264.7 cells stimulated with interferon-gamma plus lipopolysaccharide. The inhibition of NO production by the extract was due to the suppression of iNOS protein, as well as iNOS mRNA expression, determined by Western and Northern blotting analyses, respectively. In addition, the MeOH extract suppressed the production of O2- by phorbol ester stimulated RAW 264.7 cells in dose- and time-dependent manners. Collectively, these results suggest that the MeOH extract of rhizomes of C. rotundus could be developed as anti-inflammatory candidate for the treatment of inflammatory diseases mediated by overproduction of NO and O2-. PMID- 11378283 TI - Inhibitory activity of xanthine-oxidase and superoxide scavenger properties of Inga verna subsp. affinis. Its morphological and micrographic characteristics. AB - Hexane, dichloromethane and ethanolic extracts of Inga verna subsp. affinis were evaluated as inhibitors of xanthine-oxidase and as scavengers of the superoxide produced by the action of the enzyme. Ethanolic but not hexane and dichloromethane extracts showed inhibitory properties of xanthine-oxidase (IC50=27.3 microg/ml) with an additional superoxide scavenging capacity (IC50=12.7 microg/ml). The antioxidant potential was confirmed with the free radical 1,1-diphenyl-2-picryl hydrazyl (DPPH) assay, which showed that the ethanolic extract scavenges 50% DPPH free radicals at 11.6 microg/ml. HPLC study of the phenol content of the active extract, revealed the presence of ellagic and gallic acids as its main constituents. The main morphological and micrographic characteristics of Inga verna subsp. affinis are described in this paper too, in order to aid in its inequivocal identification since Inga spp. are noted for their morphological variation, which makes taxonomic classification very difficult. PMID- 11378284 TI - Study of antidiarrhoeal activity of four medicinal plants in castor-oil induced diarrhoea. AB - A study was undertaken to evaluate the effect of aqueous and methanolic plant extracts of Acorus calamus rhizome, Pongamia glabra leaves, Aegle marmelos unripe fruit and Strychnos nux-vomica root bark for their antidiarrhoeal potential against castor-oil induced diarrhoea in mice. The methanolic plant extracts were more effective than aqueous plant extracts against castor-oil induced diarrhoea. The methanolic plant extracts significantly reduced induction time of diarrhoea and total weight of the faeces. The result obtained establish the efficacy of these plant extracts as antidiarrhoeal agents. PMID- 11378285 TI - Anti-inflammatory activity of lupeol and lupeol linoleate in rats. AB - Two pentacyclic triterpenes, namely lupeol and lupeol linoleate, were investigated for their anti-inflammatory, antinociceptive, anti-pyretic and ulcerogenic properties in comparison with the commonly used non-steroidal anti inflammatory drug, indomethacin in rats. Lupeol, lupeol linoleate and indomethacin showed a reduction in paw swelling by 39, 58 and 35%, respectively, in adjuvant arthritis. Triterpenes were devoid of any antinociceptive, anti pyretic and ulcerogenic actions. However, indomethacin exhibited a positive response to these properties. These results suggest that the mechanism of action of triterpenes is different from the non-steroidal anti-inflammatory drug. PMID- 11378286 TI - Antinociceptive and antiedematogenic effects of the aqueous extract of Hyptis pectinata leaves in experimental animals. AB - The aqueous leaf extract of Hyptis pectinata (L.) Poit (Lamiaceae), popularly known in Brazil as "sambaicata" or "canudinho", was tested for its antinociceptive effects using the abdominal writhing, hot plate and formalin test models, and for its aniedematogenic effects using the carrageenin and arachidonic acid-induced rat paw edema. The aqueous extract of Hyptis pectinata administered orally at doses of 100, 200 and 400 mg/kg had a significant antinociceptive effect in the test of acetic acid-induced abdominal writhing, with 43, 51 and 54% reduction of writhes, respectively, compared to the control. An increase in hot plate latency of 47 and 37.5% was also observed in animals receiving doses of 200 and 400 mg/kg, p.o. when placed on a hot plate. In the formalin test, doses of 200 and 400 mg/kg, p.o. had no significant effect during the first phase of the test (0-5 min), while the dose of 200 mg/kg, p.o. reduced the nociceptive effect by 70% during the second phase (20-25 min). At the dose of 600 mg/kg, p.o., the aqueous extract inhibited carrageenin-induced rat paw edema by 34.1%, and the dose of 300 mg/kg administered intraperitoneally inhibited the rat paw edema induced by subplantar injection of arachidonic acid by 32.8%. These results suggest that the aqueous extract from the Hyptis pectinata leaves produces antiedematogenic and antinociceptive effects. The antinocipetion observed with the hot-plate test probably involves the participation of the opioid system. PMID- 11378287 TI - Purification and identification of active antibacterial components in Carpobrotus edulis L. AB - Very little is known about the chemical composition of Carpobrotus edulis, also known as Hotnotsfig or sourfig. However, some claims have been made in the past by traditional healers, regarding its usage as a medicinal plant. In this investigation it was initially illustrated that a crude methanolic extract of the plant exhibits strong anti-bacterial activity. Subsequently, the crude extract was fractionated by means of liquid-liquid chromatography, tannins removed by means of LH20 column chromatography and bioactive fractions with antibacterial properties isolated by means of preparative thin layer chromatography. Five bioactive compounds, individually or collectively responsible for the antibacterial property of C. edulis, were purified from an active ethyl acetate fraction. These compounds were initially identified as flavanoids using standard fingerprinting methods and eventually identified as rutin, neohesperidin, hyperoside, cactichin and ferulic acid using flavanoid standards. A sixth flavanoid with antibacterial activity was also purified but could not be identified in this way. The latter is currently isolated in larger volume for identification through nuclear magnetic resonance spectroscopy. PMID- 11378288 TI - Antifungal activity of Paraguayan plants used in traditional medicine. AB - The antifungal activity of aqueous, dichloromethane and methanol extracts from 14 Paraguayan plants used in traditional medicine for the treatment of skin diseases was assayed in vitro by the agar disk diffusion method against 11 fungal strains comprising several filamentous fungi and yeasts. Among them, the dichloromethane extracts of Acanthospermum australe, Calycophyllum multiflorum, Geophila repens and Tabebuia avellanedae, as well as the aqueous and methanol extracts of the latter, showed the highest activity. PMID- 11378289 TI - Antiplasmodial activities of some Ghanaian plants traditionally used for fever/malaria treatment and of some alkaloids isolated from Pleiocarpa mutica; in vivo antimalarial activity of pleiocarpine. AB - Fourteen Ghanaian plants used in folk medicine to treat fever/malaria were screened for activity against Plasmodium falciparum (strain K1) and were tested for general toxicity to the brine shrimp. Extracts from three of the plants, Pleiocarpa mutica, Cleistopholis patens and Uvaria chamae were found to have significant antiplasmodial activity. The extract of U. chamae was toxic to brine shrimps. These findings lend support to the use of these plants in traditional medicine. Possible toxicity due to U. chamae is a cause for concern. Five known alkaloids, pleiocarpine (1), kopsinine (2), pleiocarpamine (3), eburnamine (4) and pleiomutinine (5) were isolated from the roots of P. mutica. This is the first report of the occurrence of (4) in P. mutica. Compound (5) was the most active against P. falciparum (IC50 = 5 microM). Although (1) was inactive against malaria parasites in vitro, it was moderately active against P. berghei in mice (25 mg kg(-1) daily for 4 days reduced parasitaemia by 28.5% compared to untreated controls). PMID- 11378290 TI - Chemical composition of the essential oil from the leaves of Piper fulvescens, a plant traditionally used in Paraguay. AB - The essential oil from the leaves of the aromatic aniseed smelling plant Piper fulvescens, obtained by hydrodistillation, was investigated by GC and GC-MS, as well as by 13C-NMR after fractionation of the oil by column chromatography. A total of 37 constituents, representing 78.2% of the oil were identified. The main compounds were trans-anethole (26.4%) and ishwarane (12.1%), an unusual sesquiterpene hydrocarbon. PMID- 11378291 TI - Ginsenoside potentiates NO-mediated neurogenic vasodilatation of monkey cerebral arteries. AB - The aqueous extract of the Panax ginseng (GE) potentiated the relaxation induced by transmural electrical stimulation or nicotine in monkey cerebral arterial strips denuded of the endothelium and partially contracted with prostaglandin F(2 alpha). The response to electrical stimulation was abolished by tetrodotoxin, whereas that to nicotine was suppressed by hexamethonium. N(G)-nitro-L-arginine abolished both of the neurogenic relaxation. Atropine did not alter the potentiating effect of GE. Relaxations induced by exogenous NO were unaffected by GE. The enhancement by GE, of the neurogenic response, appears to be associated with increment in the synthesis or release of NO from the perivascular nerve. Blockade of muscarinic prejunctional inhibition, superoxide scavenging action and phosphodiesterase inhibition are not involved. PMID- 11378292 TI - Anti-diarrhoeal activity of the latex of Calotropis procera. AB - The dry latex (DL) of Calotropis procera (Asclepiadaceae), a potent anti inflammatory agent has been evaluated for anti-diarrhoeal activity. Like atropine and phenylbutazone (PBZ), a single oral dose of DL (500 mg/kg) produced a significant decrease in frequency of defecation, severity of diarrhoea and afforded protection from diarrhoea in 80% rats treated with castor oil. To understand the mechanism of its anti-diarrhoeal activity, we further evaluated its effect on intestinal transit, castor oil induced intestinal fluid accumulation (enteropooling) and electrolyte concentration in the intestinal fluid. DL produced a decrease in intestinal transit (27-37%) as compared to both normal and castor oil treated animals. Unlike atropine, DL significantly inhibited castor oil induced enteropooling. However, it did not alter the electrolyte concentration in the intestinal fluid as compared to castor oil treated rats. PMID- 11378293 TI - The aqueous extract of Rhodiola sachalinensis root enhances the expression of inducible nitric oxide synthase gene in RAW264.7 macrophages. AB - In the present study, we examined the effects of the aqueous extract of Rhodiola sachalinensis root (RSE) on the expression of inducible nitric oxide (NO) synthase (iNOS) gene in RAW264.7 macrophages. RSE synergistically increased NO synthesis in interferon-gamma-primed macrophages. Reverse transcriptase polymerase chain reaction and Northern blotting analysis revealed that RSE may provide a second triggering signal for the synergistic induction of iNOS mRNA expression. Thus, iNOS-mediated NO synthesis in response to RSE may be one mechanism whereby this herbal medicine elicits its therapeutic effects. PMID- 11378294 TI - In vitro effect of Derris scandens on normal lymphocyte proliferation and its activities on natural killer cells in normals and HIV-1 infected patients. AB - We investigated the effect of Derris scandens hydroalcoholic extract on lymphocyte proliferation, natural killer (NK) cell activity and secretion of IL-2 and IL-4. Lymphocyte proliferative response of normal individuals was significantly increased at concentrations of 10 ng/ml, 100 ng/ml, 1 microg/ml and 5 microg/ml, whereas the response was significantly decreased at 100 microg/ml. D. scandens at the concentrations of 10 ng/ml, 100 ng/ml, 1 microg/ml and 10 microg/ml significantly enhanced the function of NK cells of normal individuals. The NK cell activity of HIV-infected individuals was significantly increased at a concentration of 10 microg/ml. Furthermore, the extract was shown to induce the IL-2 secretion from normal peripheral blood mononuclear cells (PBMC), whereas the IL-4 was not induced in the presence of the D. scandens extract. Our data suggested that the hydroalcoholic extract of D. scandens possesses in vitro immunostimulating activity on human immunocompetent and immunocompromised PBMC. PMID- 11378295 TI - Otx genes in the development and evolution of the vertebrate brain. AB - Most of the gene candidates for the control of developmental programmes that underlie brain morphogenesis in vertebrates are the orthologues of Drosophila genes coding for signalling molecules or transcription factors. Among these, the orthodenticle group, including the Drosophila orthodenticle (otd) and the vertebrate Otx1 and Otx2 genes, is mostly involved in fundamental processes of anterior neural patterning. In mouse, Drosophila and intermediate species otd/Otx genes have shown a remarkable similarity in expression pattern suggesting that they could be part of a conserved control system operating in the brain and different from that coded by the HOX complexes controlling the hindbrain and spinal cord. In order to verify this hypothesis, a series of mouse models have been generated in which the functions of the murine Otx genes were: (i) fully inactivated, (ii) replaced with each other, and (iii) replaced with the Drosophila otd gene. The data obtained highlight a crucial role for the Otx genes in specification, regionalization and terminal differentiation of rostral central nervous system and lead to hypothesize that modification of their regulatory control may have influenced the morphogenesis and evolution of the brain. PMID- 11378296 TI - Developmentally regulated serotonin 5-HT2B receptors. AB - Serotonin (5-hydroxytryptamine, 5-HT) binds to numerous cognate receptors to initiate its biological effects. In this review, we have focused on the 5-HT2B receptor to address how signaling and expression of this receptor is specifically implicated in embryonic development and adult health and disease. Transduction of the 5-HT2B signaling is complex, including phospholipase C and A2 stimulation, cGMP production and a mitogenic signal that integrates the tyrosine kinase signaling pathway. Furthermore, 5-HT, through the 5-HT2B receptors, has the ability to control serotonergic differentiation of committed neuron-like cells. In addition, 5-HT2B receptors are actively involved in the transient action of 5 HT during embryonic morphogenesis. Our recent data presented the first genetic evidence that 5-HT via 5-HT2B receptors regulates cardiac embryonic development and adult functions and suggested that this receptor subtype may be involved in other physiopathological situations. In particular, 5-HT-dependent molecular mechanisms may be involved in embryonic development and postnatal maturation of the enteric nervous system. Also, the involvement of the 5-HT2B receptor in the vascular growth often observed in hypertension is likely. These probably result from reactivation of developmentally regulated receptors in pathological situations. Finally, embryonic functions of 5-HT2 receptors observed in Drosophila gastrulation suggest evolutionary conserved mechanisms. PMID- 11378297 TI - Glial cell development in Drosophila. AB - In the Drosophila central nervous system (CNS) about 10% of the cells are of glial nature. A set of molecular markers has allowed unraveling a number of genes controlling glial cell fate determination as well as genes required for glial cell differentiation. Here we focus on the embryonic CNS glia and review the recent progress in the field. PMID- 11378298 TI - Oligodendrocyte development in the spinal cord and telencephalon: common themes and new perspectives. AB - There are clear parallels between oligodendrocyte development in the spinal cord and forebrain. However, there is new evidence that in both of these regions oligodendrocyte lineage development may be more complex than we earlier thought. This stems from the recent identification of three new transcription factor genes, Olig1, Olig2 and Sox10, that are expressed from the early stages of oligodendrocyte lineage development. In this article, we highlight the common themes underlying specification and early development of oligodendrocytes in the spinal cord and telencephalon. Then, we discuss recent studies of Sox10 and the Olig genes and their implications for oligodendrocyte specification. We conclude that although the mechanisms of oligodendrogenesis appear to be fundamentally similar at different rostro-caudal levels of the neuraxis, there are still many unanswered questions about the details of oligodendrocyte specification. PMID- 11378299 TI - Role of Ca2+-stimulated adenylyl cyclases in LTP and memory formation. AB - Studies with invertebrates and vertebrates have strongly implicated the CREB/CRE transcriptional pathway in long-term memory (LTM) and transcriptionally-dependent L-LTP. It is hypothesized that LTM and L-LTP are both dependent upon a Ca2+ signal generated through activation of NMDA receptors. This review discusses evidence that Ca2+ signals generated through activation of NMDA receptors coactivate the Erk/MAP kinase and cAMP signal transduction pathways. It is hypothesized that activation of these two regulatory pathways increases the transcription of a family of genes through the CREB/CRE transcriptional pathway. Gene disruption studies have shown that Ca2+ activated adenylyl cyclases play a critical role in generating the cAMP signal required for LTM and L-LTP. Although cAMP may be required for several events in this complex signal transduction cascade, one of the major roles of cAMP may be to support nuclear translocation of Erk/MAP kinase in hippocampal neurons. PMID- 11378300 TI - Involvement of astrocytes in purine-mediated reparative processes in the brain. AB - Astrocytes are involved in multiple brain functions in physiological conditions, participating in neuronal development, synaptic activity and homeostatic control of the extracellular environment. They also actively participate in the processes triggered by brain injuries, aimed at limiting and repairing brain damages. Purines may play a significant role in the pathophysiology of numerous acute and chronic disorders of the central nervous system (CNS). Astrocytes are the main source of cerebral purines. They release either adenine-based purines, e.g. adenosine and adenosine triphosphate, or guanine-based purines, e.g. guanosine and guanosine triphosphate, in physiological conditions and release even more of these purines in pathological conditions. Astrocytes express several receptor subtypes of P1 and P2 types for adenine-based purines. Receptors for guanine based purines are being characterised. Specific ecto-enzymes such as nucleotidases, adenosine deaminase and, likely, purine nucleoside phosphorylase, metabolise both adenine- and guanine-based purines after release from astrocytes. This regulates the effects of nucleotides and nucleosides by reducing their interaction with specific membrane binding sites. Adenine-based nucleotides stimulate astrocyte proliferation by a P2-mediated increase in intracellular [Ca2+] and isoprenylated proteins. Adenosine also, via A2 receptors, may stimulate astrocyte proliferation, but mostly, via A1 and/or A3 receptors, inhibits astrocyte proliferation, thus controlling the excessive reactive astrogliosis triggered by P2 receptors. The activation of A1 receptors also stimulates astrocytes to produce trophic factors, such as nerve growth factor, S100beta protein and transforming growth factor beta, which contribute to protect neurons against injuries. Guanosine stimulates the output of adenine-based purines from astrocytes and in addition it directly triggers these cells to proliferate and to produce large amount of neuroprotective factors. These data indicate that adenine- and guanine-based purines released in large amounts from injured or dying cells of CNS may act as signals to initiate brain repair mechanisms widely involving astrocytes. PMID- 11378301 TI - Repeated prenatal corticosteroid administration delays myelination of the corpus callosum in fetal sheep. AB - Glucocorticoids regulate oligodendrocyte maturation and the myelin biosynthetic pathways. Synthetic glucocorticoids, the corticosteroids have been successfully used in clinical practice as a single course to enhance lung maturation and reduce mortality and morbidity in preterm infants with no long-term neurologic or cognitive side effects. However, a trend has arisen to use repeated courses despite an absence of safety data from clinical trials. We examined the effects of clinically appropriate, maternally administrated, repeated courses of corticosteroids on myelination of the corpus callosum using sheep as a large animal model. The corpus callosum is a major white matter tract that undergoes protracted myelination, underpins higher order cognitive processing and developmental damage to which is associated with, for example, cerebral palsy, mental retardation and attention deficit hyperactivity disorder. Pregnant ewes were given saline or betamethasone (0.5 mg/kg) at 104,111,118 and 124 days gestation, stages equivalent to the third trimester in humans. Lambs were delivered at 145 days (term), perfused and the corpus callosum examined light and electron microscopically. Total axon numbers were unaffected (P>0.05). However, myelination was significantly delayed. Myelinated axons were 5.7% in the experimental group and 9.2% in controls (P<0.05); conversely, unmyelinated axons were 88.3 and 83.7% (P<0.05). Myelinated axon diameter and myelin sheath thickness were also reduced (0.68 vs. 0.94 and 0.11 vs. 0.14 microm, P<0.05). Our data suggest that repeated prenatal corticosteroid administration delays myelination of the corpus callosum and that further safety data are needed to evaluate clinical practice. PMID- 11378302 TI - Mitogen-activated protein kinase signalling in oligodendrocytes: a comparison of primary cultures and CG-4. AB - Oligodendrocytes play a significant role in the central nervous system, as these cells are responsible for myelinating axons and allowing for the efficient conduction of nerve impulses. Therefore, any understanding we can gain about the functional biology of oligodendrocytes will give us important insights into demyelinating diseases such as multiple sclerosis, where oligodendrocytes and myelin are damaged or destroyed. Currently, much attention has focussed on the role of a family of mitogen-activated protein kinases in OL. This kinase family includes the extracellular signal-regulated protein kinases (ERKs), the stress activated c-Jun N-terminal kinase (JNK), and the 38 kDa high osmolarity glycerol response kinase (p38). The actions of mitogen-activated protein kinases in oligodendrocytes appear to range from proliferation and cell survival to differentiation and cell death. In the past, studies on oligodendrocytes have been hampered by the difficulties inherent in producing large enough quantities of these cells for experimentation. This problem arises in large part due to the post-mitotic nature of mature oligodendrocytes. Over the years, a cell line known as Central Glia-4 (CG-4) has become a popular oligodendrocyte model due to its potentially unlimited capacity for self-renewal. In this review, we will look at the suitability of the Central Glia-4 cell line as an oligodendrocyte model, specifically in respect to studies on mitogen-activated protein kinase signalling in oligodendrocytes. PMID- 11378304 TI - A tryptophan-deficient corn-based diet induces plastic responses in cerebellar cortex cells of rat offspring. AB - Sprague-Dawley male rats, fed with a tryptophan-deficient and 8% protein corn based diet were compared with a group of animals fed with 8% protein alone, and with a group fed with Chow Purina containing 23% protein. Retardation of Bergmann glial cell maturation and a concomitant retardation in granule cell migration were observed in the corn-fed group at 21 days. At 30 days of age, the dendrites of granule cells of both hypoproteic and corn-fed groups were larger than those of the Chow-fed animals. At 60 days of age, dendritic arborization of Purkinje cells was more profuse in both the hypoproteic and corn-fed rats compared with the Chow-fed group. This retardation in granule cell migration could be partially due to Bergmann glial cell immaturity. Consequently, several plastic and maybe compensatory events in both granule and Purkinje cells could have occurred, due to tryptophan deficiency resulting from the corn-based diet. PMID- 11378303 TI - Association of the HNK-1 epitope with the detergent-soluble G4 isoform of acetylcholinesterase from human neuroblastoma cells. AB - The HNK-1 carbohydrate epitope is expressed in neural and natural killer cells and is a mediator of cell adhesion. It is well documented that acetylcholinesterase has a secondary function in cell adhesion and differentiation. The presence of HNK-1 on isoforms of Torpedo and Electrophorus acetylcholinesterase, as well as isoforms from the bovine central nervous system has been described. In this paper, we have investigated the association of the epitope with acetylcholinesterase from human neuroblastoma cells. Acetylcholinesterase was extracted, with or without detergent, purified on immunoaffinity columns and the isoforms separated by sucrose density gradient sedimentation. Secreted acetylcholinesterase, from spent serum-free culture medium, was similarly treated. The presence of the HNK-1 epitope was determined by ELISA using the anti-HNK-1 and Elec 39 monoclonal antibodies. The epitope was found to be associated with the detergent-soluble G4 isoform, but not with the hydrophilic G1 nor the secreted hydrophilic G4 isoforms. Likewise, no HNK-1 was observed associated with human erythrocyte acetylcholinesterase. These results indicate that acetylcholinesterase-G4, anchored in the extracellular membrane, is capable of mediating cell-substrate adhesion through HNK-1. PMID- 11378305 TI - Differential distribution of the G protein gamma3 subunit in the developing zebrafish nervous system. AB - G proteins play an essential role in the transduction and propagation of extracellular signals across the plasma membrane. It was once thought that the G protein alpha subunit was the sole regulator of intracellular molecules. The G protein betagamma complex is now recognized as participating in many signaling events. While screening a zebrafish cDNA library to identify members of the protein 4.1 superfamily (Kelly, G.M., Reversade, B., Biochem. Cell Biol. 75 (1997), 623), we fortuitously identified a clone that encodes a zebrafish G protein gamma subunit. The 666 nucleotides of the zebrafish G protein gamma subunit cDNA encodes a polypeptide of 75 amino acids with high degree of homology to human, bovine, rat and mouse gamma subunits. BLAST search analysis of GenBank revealed that the zebrafish gamma subunit is 93% identical and 97% similar to the mammalian gamma3 subunit. The gamma3 gene was mapped to the zebrafish linkage group 21, approximately 10.76 cRays from bf, a gene with sequence homology to the human properdin factor gene. RT-PCR and in situ hybridization analyses first detected gamma3 mRNA during late somitogenesis, where it was expressed preferentially in the Vth cranial nerve, the forebrain and in ventrolateral regions of the mid- and hindbrain including the spinal cord. The ability of the zebrafish gamma3 subunit to form a signaling heterodimeric complex with a beta subunit was tested using a human beta2 subunit. The gamma3 formed a heterodimer with beta2 and the complex was capable of binding calmodulin in a calcium dependent manner. Overexpression of the beta2gamma3 complex in zebrafish embryos lead to the loss of dorsoanterior structures and heart defects, possibly owing to an up-regulation of mitogen-activated protein kinase activity and/or decline in protein kinase A signaling. Together, these data imply that a betagamma heterodimer plays a role in signal transduction events involving G protein coupled receptors and that these events occur in specific regions in the nervous system of the developing zebrafish. PMID- 11378306 TI - Effects of prenatal ionizing irradiation on the development of the ganglion cell layer of the mouse retina. AB - Prenatal exposure to ionizing irradiation has been shown to be an effective method to eliminate selectively certain neuronal population. This investigation studied the effects on the ganglion cell layer of the retinae of adult mice exposed to a gamma source (total dose=3 Gy) at 16 days gestation. There was a significant reduction in the total number of neurons (displaced amacrine+ganglion cells) in the ganglion cell layer (33%) that was mainly caused by a pronounced loss (59%) of displaced amacrine cells. The diameters of the surviving retinal ganglion cells were consistently larger than those of the controls. Prenatal irradiation is the first experimental approach that partially eliminates displaced amacrine cells. It is suggested that the morphogenesis of retinal ganglion cells may be affected by displaced amacrine cells. PMID- 11378308 TI - A MRI study of ocular hypertelorism in individuals at high risk of developing schizophrenia. PMID- 11378307 TI - Depletion of cortical target induced by prenatal ionizing irradiation: effects on the lateral geniculate nucleus and on the retinofugal pathways. AB - Studies using neonatal surgical lesions to reduce the target area of the retina have supported the idea that developing axons show only a limited specificity in their targeting. This investigation tested whether retinogeniculate axons adjust for partial target depletion by repositioning of axons. We used adult Swiss mice exposed to gamma rays at the time when layer IV cells are generated in the ventricular zone (16 days of gestation). Nissl-stained brain sections were used for histological analyses in thalamus and cortex. Retinal ganglion cells were backfilled from the optic tract with horseradish peroxidase. Intraocular injections of horseradish peroxidase were used to study the retinal projections. In the posterior cortex there was a nearly complete absence of layer IV. The irradiated animals showed a 75% reduction of the dorsal lateral geniculate nucleus. The ventral division, superior colliculus, and other visually related nuclei were not affected. The loss in the ganglion cells (15.7%) was significant but clearly smaller than that observed in the dorsal lateral geniculate nucleus (75%). Therefore, the shrinkage of the dorsal lateral geniculate nucleus led to a reduction in the area available for retinal projections. Despite partial target loss, pattern of retinal projections did not differ from that of the controls. The effect on the dorsal lateral geniculate nucleus is discussed in the light of differences between prenatal and neonatal damage of the presumptive visual cortex. The absence of aberrant retinal projections suggests that repositioning of axons is not the first mechanism employed by retinal axons to match connections in numerically disparate populations. PMID- 11378309 TI - A morphometric MRI study of the hippocampus in first-episode, neuroleptic-naive schizophrenia. AB - Magnetic resonance imaging (MRI) studies have frequently, although not unambiguously, reported hippocampal volume deficit in schizophrenia. Data on the hippocampal volumes in first-episode schizophrenia, however, are sparse. In addition, a recent topographic MRI study proposed a regionally specific volume loss in the hippocampus of chronic schizophrenics, but to date no reports have replicated this finding. In this study two-dimensional MRI-based topographic brain mapping was used to study the possibility of regional changes in the hippocampus of 22 controls and 18 patients with first-episode, neuroleptic-naive schizophrenia. Compared to controls, there were no significant differences between hippocampal volumes, regional volumes, or length of the hippocampus in the patients with schizophrenia. These data are at odds with the previous reports on hippocampal volume loss in first-episode schizophrenia, and with the hypothesis of regionally specific hippocampal volume deficit in schizophrenia. PMID- 11378310 TI - Corpus callosum area and functioning in schizophrenic patients with auditory- verbal hallucinations. AB - Auditory--verbal hallucinations (AVH) are a characteristic feature of schizophrenia. Patients with AVHs have been found to differ from non hallucinating patients in volumes of certain asymmetrical brain structures on MRI, and on certain neuropsychological measures. There is also evidence of corpus callosum (CC) abnormalities in schizophrenia, and it has been proposed that abnormalities of inter-hemispheric transmission may underlie hallucinations and other symptoms. The aim of this study was to examine whether patients with AVHs have smaller corpora callosa than those without AVH, and whether CC size is related to performance on neuropsychological tests of functional cerebral asymmetry. Seventy-one DSM-IV male schizophrenics were recruited on the basis of their hallucination history plus 33 matched normal controls. Twenty-nine patients had no history of AVH, and 42 had a strong history of AVH. The mid-sagittal surface area and longitudinal length of the CC were measured from T(1)-weighted spin echo images. Callosal area was divided into four sections. There were no significant differences in any of the measurements between the two patient groups, or between patients with schizophrenia and controls. There was no association between CC measures and handedness, or performance on dichotic listening or finger tapping tasks. The results of this study do not lend support for there being a major morphological abnormality of the corpus callosum in schizophrenic patients, or for a specific relationship to AVH. However, a significant association between CC area and overall grey and white matter volumes was noted in the hallucinating patients and, to a lesser extent, in the non hallucinators, which may point to differing influences on brain development or degeneration in such patients compared with normal controls. PMID- 11378311 TI - Cerebral gray and white matter reductions and clinical correlates in patients with early onset schizophrenia. AB - Few magnetic resonance imaging studies of schizophrenia have investigated brain tissue volumes and their relation to clinical symptoms in patients with an early age at illness onset. The twofold purpose of the study was to investigate both gray and white matter volumes in schizophrenic men with an early age at illness onset, and to determine whether clinical features correlated with tissue volume changes, using an automated voxel-by-voxel image analysis procedure. Twenty male patients with DSM-IV diagnoses of schizophrenia, and an early age at onset (m+/ SD=19+/-2) were compared with 20 age-matched health men. Magnetic resonance (1.5 T) scans were obtained with an Inversion-Recovery prepared fast gradient echo sequence enhancing gray and white matter contrast. Statistical Parametric Mapping was used for image segmentation and comparison. Patients had significant gray matter reductions in medial frontal gyri, left insula, left parahippocampus, and left fusiform gyrus; bilateral white matter reductions in frontal lobes, and increased total cerebrospinal fluid volume were also observed. Negative symptom scores were negatively related to white matter volumes in cingulate regions, and in the right internal capsule. These findings emphasize a pattern of left hemisphere gray matter abnormalities, and suggest that fronto-paralimbic connectivity may be altered in men with early onset schizophrenia. PMID- 11378312 TI - Differential activation of temporal cortex during sentence completion in schizophrenic patients with and without formal thought disorder. AB - The neural correlates of processing linguistic context in schizophrenic patients with formal thought disorder (FTD) were examined. Six right-handed male patients with prominent 'positive' FTD were compared with six schizophrenic patients without FTD and seven volunteers, matched for cognitive and demographic variables. Functional magnetic resonance imaging (IMRI) was used to measure cerebral activation while subjects read and completed sentence stems out loud. During a GENERATION condition, subjects were required to generate a word which completed the sentence stem appropriately. During a DECISION condition, subjects selected and articulated one of two presented terminal words. A READING condition served as baseline. The three conditions were compared with each other. Regions activated were identified in each group, and between-group differences were detected using an ANCOVA. When GENERATION was compared with READING, FTD patients showed less activation in the right superior temporal gyrus than patients without FTD or controls, but greater activation in the left inferior frontal, inferior temporal and fusiform gyri. FTD patients also showed an attenuated right temporal response when GENERATION was compared with DECISION. This differential engagement of the right temporal cortex was independent of differences in the speed or accuracy of responses, whereas the left fronto-temporal differences in activation were not evident after covarying for task errors. The attenuated engagement of right temporal cortex, which is implicated in language comprehension at the discourse level, is consistent with neuropsychological evidence linking thought disorder with deficits in processing linguistic context. PMID- 11378313 TI - Quantitative EEG in schizophrenia and in response to acute and chronic clozapine treatment. AB - Topographic quantitative electroencephalographic (EEG) power and frequency indices were collected in 17 treatment refractory, DSM-III diagnosed schizophrenic patients, before and after acute (single dose) and chronic (six weeks) clozapine treatment, as well as in 17 healthy volunteers. Prior to treatment, patients exhibited greater overall absolute theta power, slower mean alpha frequency and elevated absolute delta and total power in anterior regions. Acute dosing increased total spectrum power globally, slow wave power posteriorally, mean alpha frequency and beta power anteriorally and decreased alpha power posteriorally. Six weeks of clozapine treatment significantly reduced clinical ratings of positive and negative symptoms as well as symptoms of global psychopathology. Chronic treatment resulted in EEG slowing as shown by decreases in relative alpha power, mean beta/total spectrum frequency and by widespread increases in absolute total and delta/theta power. The preliminary findings suggest that brain electric profiling may be a promising tool for assessing and understanding the central impact of pharmacotherapeutic interventions in schizophrenia. PMID- 11378314 TI - Decreased functional connectivity of EEG theta-frequency activity in first episode, neuroleptic-naive patients with schizophrenia: preliminary results. AB - We explored and refined the hypothesis that during a first episode of acute schizophrenia a disorganization of brain functioning is present. A novel EEG measure was introduced, Global Field Synchronization (GFS), that estimates functional connectivity of brain processes in different EEG frequency bands. The measure was applied to EEG's from 11 never-treated, first-episode, young patients with an acute, positive, schizophrenic symptomatology and from 19 controls, residing in Bern, Switzerland. In comparison to age- and sex- matched controls, patients had significantly decreased GFS in the theta EEG frequency band, indicating a loosened functional connectivity of processes in this frequency. The result was confirmed in an independent, comparable patient group from Osaka, Japan (9 patients and 9 controls), thus making a total of 20 analyzed patients. Previous EEG research in healthy, awake subjects indicated a positive correlation of theta activity with memory functions. Thus, our result suggests a loss of mutual interdependence of memory functions in patients with acute schizophrenia, which agrees well with previous reports of working memory dysfunction in schizophrenia. PMID- 11378315 TI - Auditory verbal hallucinations and dysfunction of the neural substrates of speech. AB - OBJECTIVE: to evaluate the neural substrate of auditory verbal hallucinations (AVH), the correlation between AVH and subvocal speech (hereafter SVS), and the relationship between speech and AVH. METHOD: we reviewed the papers found by an electronic literature search on hallucinations and speech. The review was extended to the papers cited in these publications and to classical works. RESULTS: there is no conclusive evidence of structural abnormality of the speech perception area in hallucinating schizophrenic patients. However there is evidence of electrophysiological abnormalities of the auditory and speech perception cortices. Functional imaging data are inconsistent, yet point to the left superior temporal gyrus as one of the neural substrates for AVH. There is also evidence that SVS could accompany the experience of AVH. CONCLUSION: there is evidence that dysfunction of brain areas responsible for speech generation is a fundamental mechanism for generating AVH in schizophrenia. It results in a secondary activation of Wernicke's area (speech perception) and Broca's area (speech expression). The first leading to the experience of hallucinations, and the second, eventually, gives rise to a variable degree of vocal muscle activity detectable by EMG, and/or faint vocalizations detectable by sensitive microphones placed at proximity of the larynx. Direct stimulation or disease of Wernicke's area produces AVH without SVS. PMID- 11378316 TI - Improvement of schizophrenic patients' subjective well-being under atypical antipsychotic drugs. AB - Recent research indicates that subjective well-being is a major determinant of medication compliance in schizophrenia. However, it is yet unresolved whether atypical neuroleptics differ regarding subjective side-effects. A self-report instrument has been constructed to evaluate 'subjective well-being under neuroleptics' (SWN). The primary aims of the present study were to develop a short form of the SWN and to investigate the extent to which the atypical antipsychotic improves the patient's subjective well-being. The short form of the SWN was constructed following an item analysis based on data from 212 schizophrenic patients medicated with either typical or atypical antipsychotics. The short form of the SWN showed sufficient internal consistency and good construct validity. The SWN was only moderately correlated with positive and negative syndrome scale (PANSS) scores or changes in psychopathology (r=-0.20 to 0.37). SWN-ratings in patients receiving olanzapine were superior compared to those of patients medicated with either clozapine or risperidone on three of five domains of well-being. Clozapine reduced global psychiatric symptoms significantly more than risperidone. It is concluded that the assessment of subjective well-being under antipsychotic treatment provides an independent outcome measure which is relevant to compliance. PMID- 11378317 TI - Risperidone versus haloperidol in long-term hospitalized chronic patients in a double blind randomized trial: a post hoc analysis. AB - BACKGROUND: Patients who remain in hospital for an extended time pose a special therapeutic challenge. OBJECTIVES: The goal of this study was to examine whether the acute response of long-term hospitalized schizophrenic patients differs between haloperidol and risperidone based on a post hoc, sub-analysis of data from a large double blind pivotal trial. METHOD: Data on chronic schizophrenic patients who had been hospitalized for at least 60 days (median 351 days) prior to entering this 8-week randomized double blind controlled trial were examined. This included 75 patients treated with 4 mg of risperidone and 69 treated with 10mg of haloperidol. Changes in symptoms were assessed with the Positive and Negative Syndrome Scale for Schizophrenia (PANSS) and the Clinical Global Impression (CGI). Data were analyzed using analysis of variance. RESULTS: The analyses revealed that patients receiving risperidone improved significantly more than those treated with haloperidol. CONCLUSIONS: Results suggest that the most often prescribed dose of risperidone, 4 mg, might be more effective for long-stay chronic schizophrenic patients than haloperidol 10mg. PMID- 11378318 TI - Relationship of the factor structure of psychopathology in schizophrenia to the timing of initial intervention with antipsychotics. AB - Timing of intervention with antipsychotic medication may influence long-term outcome in schizophrenia in a manner that is poorly understood. This study evaluated psychopathology, its factor structure, and cognitive dysfunction in older patients with chronic schizophrenia in relation to the intervals from onset of psychosis to initiation of treatment with antipsychotics, and from initiation of antipsychotic treatment to current assessments. The subjects were 129 patients with schizophrenia, many of whom became ill in the preneuroleptic era. Their current psychopathology was assessed using the Positive and Negative Syndrome Scale, and its factor structure examined using principal component analysis. Current general and executive cognitive function was evaluated using the Mini Mental State Examination and the Executive Interview, respectively. Using multiple regression modelling, increasing duration of initially unmedicated psychosis, but not the much longer duration of subsequently treated illness, was the primary predictor of psychomotor poverty (negative symptoms) but not of reality distortion or disorganisation over the three domains of psychopathology resolved; duration of initially unmedicated psychosis marginally predicted the severity of general, but not of executive, cognitive dysfunction. Delayed intervention with antipsychotics appears associated with poorer long-term course in terms of increased severity of psychopathology in the psychomotor poverty domain. PMID- 11378319 TI - CASIG: a consumer-centered assessment for planning individualized treatment and evaluating program outcomes. AB - This paper reports the psychometric characteristics of a measure that assesses the treatment outcomes of individuals with serious and persistent mental illness. Unlike other outcome measures, this one is designed to be embedded in the clinical process of planning and evaluating treatment. It collects individualized data, structures how the data are used to plan and evaluate a client's treatment, and produces aggregate information relevant for research and program purposes. Two parallel versions were developed: one for the client's self-report, and one for an informant's report. The self-report measure was administered by peer interviewers to 244 community interviewees, and by inpatient peer-interviewers to 93 inpatient interviewees. The community interviewees also completed the BASIS-32 and SF-36. Informants for 103 of the community interviewees completed the informant version of the measure, and the CCAR. Inpatient staff completed the informant version for 161 inpatient residents without regard for matching the 93 inpatient interviewees. The two versions had acceptable internal consistency, test--retest, and interrater reliabilities. Correlations of the community interviewees' and informants' results with the BASIS-32, SF-36, and CCAR provided evidence of convergent and discriminant validity, as did contrasts between community and inpatients interviewees. The usefulness of the instrument for clinical, program and research purposes is discussed, with emphasis on the characteristics that enhance its value in clinical practice --- assessment of meaningful outcomes, operationalization of client empowerment, comprehensiveness, easy administration, and continuity across time and provider. Also discussed is a computer-based program to summarize and present the results in a rapid, clinically meaningful manner. PMID- 11378320 TI - Diagnosis and treatment of psychiatric disorders in children with a schizophrenic parent. AB - Many children of a schizophrenic parent may inherit a portion of the genetic risk factors for schizophrenia. The frequency of DSM-IV psychopathology in children of a schizophrenic parent and the frequency and type of mental health treatment accessed by these youths is not well understood. Twenty-eight adults with schizophrenia were identified and 43 of their 6--15 year old children recruited. Clinical diagnoses, based on a structured DSM-IV interview; severity of impairment, based on the Child Global Assessment Scale; and treatment histories were obtained. Seventy-four per cent of children-with-a-schizophrenic-parent met diagnostic criteria for a current Axis I psychiatric disorder. The most common diagnostic categories included attention deficit/hyperactivity disorder (40%), any anxiety disorder (23%), and any depressive disorder (12%). Psychosis was present in 9% of this childhood sample. Of those children with a psychiatric diagnosis, 47% demonstrated current moderate or severe impairment. Approximately half of the children had received mental health evaluations and 26% had experienced at least one psychiatric medication trial. Children-with-a schizophrenic-parent have frequent, often impairing, psychiatric problems. Despite this high prevalence, mental health evaluation and treatment is of similar frequency and type to other at-risk populations. The effectiveness and appropriateness of standard treatments remain unstudied in children-with-a schizophrenic-parent. PMID- 11378321 TI - Symptom dimensions in acutely ill patients with affective, schizoaffective, and schizophrenic disorders. PMID- 11378322 TI - Successful outcome using quetiapine in a case of treatment-resistant schizophrenia with assaultive behavior. PMID- 11378323 TI - Cytokine mRNA profiles in cultured human skin component cells exposed to various chemicals: a simulation model of epicutaneous stimuli induced by skin barrier perturbation in comparison with that due to exposure to haptens or irritant. AB - The skin protects our body by producing an efficient barrier membrane, the stratum corneum, from desiccation as well as from various damaging effects of environmental chemicals. Although the skin expresses various cytokines after barrier perturbation, exact cell types producing each cytokine have not been determined. Using a cell culture system, we analyzed the initial responses of various cutaneous cells to treatments simulating epicutaneous stimuli induced by a barrier perturbation of the skin in comparison with those caused by irritant or hapten exposure. We used cultured normal human epidermal keratinocytes (NHEK), human microvascular endothelial cells (HMVEC) and normal human dermal fibroblasts (NHDF). We treated them with the following chemicals and examined their cytokine mRNA levels 6 h later: high osmotic (0.5 molar) NaCl and hydrogen peroxide (H2O2), which simulate desiccation and exposure to high oxygen pressure, respectively, that may take place in vivo after perturbation of the barrier. In addition, we also studied their response to two representive haptens, nickel chloride (NiCl2) and dinitrochlorobenzene (DNCB), and an irritant, sodium dodecyl sulfate (SDS). We found that 0.5 M NaCl treatment increased mRNA levels of proinflammatory cytokines such as IL-1alpha, IL-6 and IL-8 as well as ICAM-1 in NHEK and IL-1alpha, IL-1beta and IL-6 mRNA levels in NHDF. In contrast, H2O2 treatment remarkably increased IL-10, GMCSF and ICAM-1 mRNA levels in NHEK, and IL-6 mRNA levels in HMVEC and NHDF. The exposure to haptens did not induce any remarkable increase in mRNA levels of the proinflammatory cytokines in NHEK. But NiCl2 increased IL-1alpha, IL-6 and IL-8 mRNA levels in HMVEC, while DNCB increased only their IL-6 mRNA levels. By contrast, SDS stimulated all the cell types to increase at least some of these proinflammatory cytokine mRNA levels. Our present data suggest that each skin component cell participates in inflammatory processes of the skin through its distinctive cytokine production profile when the skin barrier is compromized physically or chemically. PMID- 11378324 TI - Relationship between lymphocyte cyclosporin sensitivity and clinical progress of psoriasis. AB - Cyclosporin (CYA) is a therapeutic agent used in the treatment of psoriasis vulgaris. However, the effectiveness of CYA therapy varies among patients. In the present study, due to the fact that CYA mainly acts on lymphocytes, we hypothesized that a measurement of the sensitivity of lymphocytes to CYA in vitro (IC50) could be applied to patients with psoriasis vulgaris to determine therapeutic success. We measured IC50 levels of 32 patients presenting with psoriasis prior to CYA administration, and classified them into three groups according to IC50 levels: favorable, moderate and low sensitivity. CYA sensitivity levels were correlated with the degree of improvements in psoriasis area and severity index (PASI) scores, CYA dosage and occurrence of side-effects (hepatopathy and nephropathy). Results showed the degree of improvement in PASI scores differed significantly between the favorable and low sensitivity groups (P < 0.05). Furthermore, CYA dosage was lowest in the favorable sensitivity group and highest in the low sensitivity group. Moreover, hepatopathies and nephropathies were detected in the low and moderate sensitivity groups, but not in the favorable sensitivity group. These results suggest that the effectiveness of CYA therapy as a treatment of psoriasis vulgaris is affected by the sensitivity of lymphocytes in each patient. PMID- 11378325 TI - Neuron-specific PGP9.5 expression in rat hair follicle development and cycle. AB - Protein gene product PGP9.5 is a neuron-specific ubiquitin C-terminal hydrolase. We found that it also has immunoreactivity in the hair follicle of the Wistar rat dorsal skin and its expression patterns change with the development and cycle. During the morphogenesis, the PGP9.5 was expressed in the hair germ and hair peg elongated from the epidermis, and became restricted in the outer root sheath as the development progressed. In catagen, however, the PGP9.5 was detected in the tailing epithelial strand of the regressing proximal follicle epithelium, and in the keratinocytes directly contacted with the club hair, but rarely in the outer root sheath. With the beginning of the anagen of the second hair follicle, the PGP9.5 was again expressed in the second hair germ, and in the keratinocytes surrounding the remaining club hair and of distal follicle of the first hair. These findings showed that PGP9.5 is not specific to the neuron but is also involved in the hair follicle, and should provide new insight into the development and regression of the hair follicle. PMID- 11378326 TI - Role of stem cell factor and monocyte chemoattractant protein-1 in the interaction between fibroblasts and mast cells in fibrosis. AB - Mast cell infiltration and accumulation is known to occur in tissue fibrosis. Increased numbers of mast cells are detected in scleroderma or hypertrophic scar skin, however, neither the role of mast cells nor the interaction of fibroblasts and mast cells in fibrosis are fully understood. A growing body of evidence indicate that mast cells are rich source of cytokines, growth factors or chemokines, which are suggested to play an important role in the induction of fibrosis. Recent in vivo and in vitro studies suggest the involvement of monocyte chemoattractant protein-1 (MCP-1), a member of the C-C chemokine family, in fibrosis. Here, we examined the effect of stem cell factor (SCF), a mast cell growth factor, on MCP-1 gene expression in a human mast cell line, HMC-1, and as well as the effect of MCP-1 on alpha1(I) collagen gene expression in human skin fibroblasts. HMC-1 cells spontaneously expressed MCP-1 mRNA transcripts, which was detectable by in situ hybridization and Northern blot analysis. Stimulation with SCF further upregulated MCP-1 mRNA expression in a time- and dose-dependent manner, and stimulation with 100 ng/ml SCF for 24 h induced a 3-fold increase of MCP-1 mRNA expression in HMC-1 cells as compared with unstimulated cells. The concentration of MCP-1 protein in the culture supernatants of 50 ng/ml SCF stimulated HMC-1 cells (3816+/-70 pg/ml) was significantly elevated compared to unstimulated cells (2588+/-130 pg/ml) (P < 0.01), as assessed by ELISA. Adversely, MCP-1 induced alpha1(I) collagen mRNA expression in normal skin fibroblasts dose-dependently. Finally, comparative study revealed that the concentration of SCF in the culture supernatants of scleroderma fibroblasts at primary passages was significantly increased (344.6+/-182.4 pg/ml), as compared with normal skin fibroblasts (72.4+/-20.2 pg/ml) (P<0.05). These results suggest that fibroblast-derived SCF upregulates MCP-1 expression and synthesis in mast cells, which acts on fibroblasts to enhance alpha1(I) collagen mRNA expression. Our data may indicate an important interaction of fibroblasts and mast cells, via SCF and MCP-1, in the induction of fibrosis. PMID- 11378327 TI - Localized heat urticaria: a clinical study using laser Doppler flowmetry. AB - We studied the pathophysiology of localized heat urticaria using laser Doppler flowmetry (LDF) in two patients with this rare disease. In heat challenge tests, performed with different challenge times and temperatures, a heat stimulator with a thermoregulated metal disc was utilized. Immediately after removal of the heat source, cutaneous blood flow (CBF) changes in the tested sites were monitored with LDF. In both patients the increase in (CBF) took place at some intervals after a heat challenge, synchronous with the start of the urticarial response. This interval, or the latency time (LT), showed distinct inverse proportion to the intensity of heat stimuli and was prolonged by effective treatments, such as application of antihistamines and repeated heat exposure by LDF. Therefore, the time of latency might be regarded as a good indicator of the severity of illness and therapeutic effectiveness, and thus might reflect the relationship between the degree of heat stimuli and the releasing process of chemical mediator(s) in patients with localized heat urticaria (LHU). PMID- 11378328 TI - Interleukin-8-positive neutrophils in psoriasis. AB - We performed an immunohistochemical study to try to determine the cellular source of interleukin-8 (IL-8) in psoriatic skin lesions. IL-8 was positively stained in the vast majority of neutrophils but not in the mononuclear cells, macrophages, or keratinocytes. IL-8-positive neutrophils were seen both in Munro's microabcesses in cases of psoriasis vulgaris and in a small spongiform pustule and much larger macropustules of Kogoj in cases of pustular psoriasis. Some IL-8 positive neutrophils were observed in the upper dermis of pustular psoriasis. The staining was considered to be specific because it could be completely blocked by preabsorption with recombinant IL-8. In addition, stimulation of human neutrophils with lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF alpha) for 18 h induced IL-8 production in vitro. In our study, IL-8 was expressed in the neutrophils of psoriasis, suggesting that neutrophils are one of the sources of IL-8 in psoriasis. The expression of IL-8 and the influx of neutrophils led us to speculate that the IL-8 autocrine and/or paracrine system functions in the formation of the microabcesses and pustules in proriasis. PMID- 11378329 TI - Characterization of mutations of the type VII collagen gene (COL7A1) in recessive dystrophic epidermolysis bullosa mitis (M-RDEB) from three Korean patients. AB - In recent years, the molecular basis for the main subtypes of epidermolysis bullosa (EB) has been elucidated with pathogenetic mutations delineated in ten different genes encoding structural components of the dermal-epidermal junction. Both the autosomal dominant and recessive forms of dystrophic EB (DEB) is caused by mutations in the COL7A1 gene. Type VII collagen is a major component of anchoring fibrils, structural elements that stabilize the attachment of the basement membrane to underlying dermis. Recent delineation of the exon-intron organization of the COL7A1 gene provided the basis for the comprehensive design of PCR primer pairs that amplified exons in genomic DNA by placing the primers on the flanking introns. A number of COL7A1 mutations have been reported and some genotype-phenotype correlations are starting to emerge. In this study, we examined mutational analyses from three Korean patients with recessive dystrophic EB (RDEB) mitis. We designed and optimized primers according to the previously reported sequences. Such PCR amplification products can be examined by electrophoretic scanning technique, CSGE heteroduplex analyses. Utilizing heteroduplex analyses, we have identified a number of sequence variants in COL7A1 both in unaffected individuals and in patients with M-RDEB. Mutation detection of the COL7A1 gene revealed six allelic mutations (V6677E, P6685S, Y3749S, P6084S, P6695R and G6697C). We suggest that the full length of type VII collagen polypeptide are synthesized, but those missense mutations, that may affect a critical amino acid, can alter the conformation of the protein and interferes with the assembly and packing of type VII collagen molecules into anchoring fibrils. Immunohistochemical study of skin biopsies by use of anti-type VII collagen antibody showed markedly reduced staining and presence of a dermo/epidermal cleavage. This is the first report of a COL7A1 mutation study in DEB from Korean patients. We hope that these data contribute to the expanding database on COL7A1 mutations in dystrophic epidermolysis bullosa, and further illustrate the extensive diversity of mutational events that led to the RDEB phenotype. PMID- 11378330 TI - Expression of monocyte chemoattractant protein-1 in the lesional skin of systemic sclerosis. AB - Systemic sclerosis (SSc) is a connective tissue disease with unknown etiology characterized by excessive deposition of collagen in the skin as well as various internal organs. One of the characteristic histological features is the presence of infiltrating mononuclear cells in the dermis in its early stage. As well as T cells, macrophages are implicated to play an important role in the initial pathologic changes associated with SSc by releasing fibrogenic cytokines, including transforming growth factor-beta or platelet-derived growth factor. However, the precise mechanism for increased monocyte/macrophage recruitment in the lesional skin of SSc is still not completely elucidated. Monocyte chemoattractant protein-1 (MCP-1) is a predominant monocyte chemoattractant secreted by various cells types including mononuclear cells, fibroblasts, smooth muscle cells, endothelial cells, or keratinocytes. In this study, we examined the expression of MCP-1 protein and mRNA in the lesional skin of seven patients with SSc by immunohistochemistry and in situ hybridization. Results of immunohistochemistry showed that MCP-1 was detected on infiltrating mononuclear cells and fibroblastic cells in scleroderma skin, whereas normal skin showed only minimal MCP-1 expression. We demonstrated the expression of MCP-1 mRNA in infiltrating mononuclear cells and keratinocytes in scleroderma and contact dermatitis skin. In addition, signals were also detected in fibroblasts in the lesional skin of scleroderma, whereas fibroblasts in normal skin and contact dermatitis skin did not express MCP-1 mRNA. These findings suggest that MCP-1 plays a role in recruiting monocyte/macrophages in the lesional skin of scleroderma and that activated fibroblasts in scleroderma are involved in this process. PMID- 11378331 TI - RCAS1 antigen is highly expressed in extramammary Paget's disease and in advanced stage squamous cell carcinoma of the skin. AB - Receptor-binding cancer antigen expressed on SiSo cells (RCAS1), which is a type II membrane protein expressed on cervical carcinoma cells, induces apoptosis in RCAS1 receptor expressing cells. RCAS1 is thus presumed to protect tumor cells from immune surveillance by infiltrating RCAS1 receptor-positive immunocytes (Sonoda et al. Int J Oncol 1995; 6: 1899-1904; Nakashima et al. Nature Med 1999; 5: 938-942). We performed immunohistochemical analysis of RCAS1 expression in various skin tumors. RCAS1 was not detected in normal human epidermis. One of 21 seborrheic keratosis (4.8%), one of 12 actinic keratosis (8.3%), two of 16 keratoacanthomas (12.5%), and two of 14 basal cell carcinomas (14.2%) expressed RCAS1. RCAS1 was not detected in Bowen's disease (0/17). RCAS1 was positive in 45 of 61 (73.8%) squamous cell carcinomas. Interestingly, the expression of RCAS1 was mostly correlated with clinical stages of squamous cell carcinoma. It was found that 46.1% of stage I, 61.1% of stage II, 85.7% of stage III, and 83.3% of stage IV squamous cell carcinomas were RCAS1-positive. In addition, RCAS1 was found to be highly expressed in extramammary Paget's disease. Fifty nine of 63 extramammary Paget's disease samples (93.7%) were positive for RCAS1. Fifty eight (92%) showed co-expression of RCAS1 and carcinoembryonic antigen (CEA). While two of 24 cases of melanoma (8.3%) expressed RCAS1 antigen, none of 20 cases of nevus pigmentosus showed positive staining. These results indicate that RCAS1 is a highly sensitive marker for extramammary Paget's disease. RCAS1 is also expressed in various skin tumors including squamous cell carcinoma, where positive correlation with clinical staging was documented. PMID- 11378332 TI - Genotypic and phenotypic characterization of Staphylococcus aureus strains isolated in subjects with atopic dermatitis. Higher prevalence of exfoliative B toxin production in lesional strains and correlation between the markers of disease intensity and colonization density. AB - Staphylococcus aureus strains generally colonize eczematous lesions of subjects with atopic dermatitis much more frequently than in the skin of normal individuals. The aim of this study was to provide a detailed genotypic and phenotypic analysis of S. aureus strains colonizing four different sites (lesional and non-lesional skin areas, nasal and pharyngeal mucosas) of 49 patients with atopic dermatitis. The 88 isolates were analyzed in duplicate by pulsed field gel electrophoresis and in their exfoliative toxin A or B production by latex test. The patients were characterized by age, sex, severity scoring of atopic dermatitis and serum eosinophil cationic protein. Fourteen (28.6%) of the patients were completely negative for S. aureus while 35 (71.4%) were positive in at least one site. The severity scores and eosinophil cationic protein levels were significantly correlated variables (P<0.001), linked to the colonization intensity (P ranging between 0.05 and <0.001 depending on the site) and to the number of colonized sites (P at least <0.01). The genotypic patterns, widely heterogeneous, showed no restriction to peculiar patterns. Only eight strains produced exfoliative toxin B which was significantly restricted to the lesional isolates (P=0.012). PMID- 11378334 TI - Why remove four by chance when one will suffice? PMID- 11378333 TI - Up-regulation of putative hyaluronan synthase mRNA by basic fibroblast growth factor and insulin-like growth factor-1 in human skin fibroblasts. AB - Recently, cDNAs for the three putative human hyaluronan synthase (HAS) genes, HAS1, HAS2 and HAS3, have been cloned. In this study we investigated the effects of basic fibroblast growth factor (bFGF) and insulin-like growth factor-1 (IGF-1) on the expression of HAS genes in cultured skin fibroblasts. Northern blot analyses showed that treatment of fibroblasts with bFGF enhanced the mRNA levels of all three genes. HAS2 gene expression showed the strongest up-regulation with a more than 10-fold increase at 50 ng/ml of bFGF. bFGF also increased hyaluronan production. Incubation of fibroblasts with IGF-1 increased HAS1, HAS2, and HAS3 mRNA levels, as well as hyaluronan production. Our results suggest that up regulation of the HAS genes by bFGF and IGF-1 is closely associated with the stimulation of hyaluronan synthesis, and that effects of growth factors on HAS gene expression may have important implications for tissue remodeling, such as in development and wound healing. PMID- 11378335 TI - End of life care in patients with malignant disease. PMID- 11378336 TI - Sentinel node biopsy for breast cancer may have little to offer four-node samplers. results of a prospective comparison study. AB - The aims of the study were to determine how often four node axillary sampling (4NAS) encompasses the sentinel node (SN) and to compare the relative sensitivity of 4NAS with sentinel node biopsy (SNB) for axillary node staging. 200 patients with breast cancer were preoperatively injected with 27 MBq 99m-Tc-labelled colloid adjacent to the tumour. At operation, standard 4NAS was performed. Each node was counted ex vivo using a probe. A search was then made to find a node with higher counts in vivo directed by the probe. If found, it was excised. Each node was submitted separately to pathology. A SN was identified in 191 patients (96%). The SN was contained in the 4NAS in 153 patients (80%) and identified separately in 38 patients (20%). Of 60 node-positive patients, 49 were positive by 4NAS and SNB, the SN was not identified in 2 and in 8 the SN was falsely negative compared with 4NAS. For 1 patient, the SN was positive and the 4NAS negative. SNB performed using radiolabelled colloid has no advantage over 4NAS when nodes are assessed by standard histological technique. PMID- 11378337 TI - Deriving a compound quality of life measure from the EORTC-QLQ-C30/LC13 instrument for use in economic evaluations of lung cancer clinical trials. AB - Many clinical trials involve parallel collection of quality of life (QoL) and economic data, requiring patients to complete similar questionnaires at regular intervals. This additional burden often leads to disappointing response rates and inconclusive results. Data obtained in the LU-16 trial with the European Organization for Research and Treatment of Cancer Quality of Life (EORTC-QLQ C30)/LC-13 QoL instrument for lung cancer were re-analysed, using multivariate techniques. The analysis demonstrated the inherent non-linearity of QoL data, with resulting interpretational problems. A new integrated linear QoL measure was developed which maximises the use of the information collected and can serve as a proxy utility measure for economic evaluation. It was successfully validated with data from another lung cancer trial with encouraging results. For individual patients, trends in QoL are revealed more clearly with narrower confidence intervals. This approach yields relative weightings and rankings for the main issues affecting QoL ratings in lung cancer patients, most importantly fatigue, breathlessness, poor concentration and disruption to family and social life. PMID- 11378338 TI - Overexpression of c-erbB2 protein correlates with disease-stage and chromosomal gain at the c-erbB2 locus in non-small cell lung cancer. AB - Overexpression of the c-erbB2 protein is observed in a variety of malignancies including non-small cell lung cancer (NSCLC). We aimed to determine the rate of c erbB2-overexpression in our tumour collection and to clarify its correlation with the chromosomal status at the c-erbB2 locus 17q21 in NSCLC. Eighty-nine NSCLC were analysed immunohistochemically using a polyclonal c-erbB2 antibody (DAKO). The staining was scored according to the guidelines of the Clinical Trial Assay recommendations (0-3+). Of these, 44 cases were also analysed by comparative genomic hybridisation (CGH). Overexpression was observed in 37% of the cases (score>1) which was associated with higher disease stages and a positive nodal status in adenocarcinomas. Chromosomal gains at 17q21 were clearly correlated with overexpression of the gene (P=0.009). In addition, there was a highly significant correlation between the c-erbB2 expression comparing the whole section immunostaining analysis and a 127 lung tumour tissue array which included 74 of the 89 cases that were analysed by the classical procedure. We conclude that c-erbB2 is a marker of tumour progression in NSCLC which can be observed on protein level and reflects chromosomal alterations at 17q21. PMID- 11378339 TI - A randomised phase II study on neo-adjuvant chemotherapy for 'high-risk' adult soft-tissue sarcoma. AB - The aim of this study was to examine the strategy, feasibility and outcome of neo adjuvant chemotherapy, with doxorubicin and ifosfamide, in adult patients with 'high-risk' soft-tissue sarcomas. Patients with 'high-risk' soft-tissue sarcomas, defined as tumours > or =8 cm of any grade, or grade II/III tumours <8 cm, or grade II/III locally recurrent tumours, or grade II/III tumours with inadequate surgery performed in the previous 6 weeks and therefore requiring further surgery, were randomised between either surgery alone or three cycles of 3-weekly doxorubicin 50 mg/m(2) intravenous (i.v.) bolus and ifosfamide 5 g/m(2) (24 h infusion) before surgery. The type of surgery had to be planned at randomisation. Tumours were to be amenable to surgery by amputation, compartmental resection, wide or marginal excision. If chemotherapy was given, surgery had to be performed within 21 days after the last chemotherapy. Patients received postoperative radiotherapy in cases of marginal surgery, microscopically incomplete resection and no further possibility for surgery, and in cases of surgery because of local recurrence. 150 patients were entered into the study and 134 were eligible, 67 in each arm. The most frequent side-effects of chemotherapy were alopecia, nausea and vomiting (95%), and leucocytopenia (32%). One patient died of neutropenic fever after the first cycle of chemotherapy. Chemotherapy did not interfere with planned surgery and did not affect postoperative wound healing. Limb-salvage was achieved in 88%, amputation was necessary in 12% (all according to the plan at randomisation). The trial was closed after completion of phase II, since accrual was too slow to justify expanding the study into the scheduled phase III study. At a median follow-up of 7.3 years, the 5 year disease-free survival is estimated at 52% for the no chemotherapy and 56% for the chemotherapy arm (standard error: 7%) (P=0.3548). The 5 year overall survival for both arms is 64 and 65%, respectively (standard error 7%) (P=0.2204). Neo-adjuvant-chemotherapy with doxorubicin and ifosfamide at these doses and with this schedule was feasible and did not compromise subsequent treatment, surgery with or without radiotherapy. Although not powered to draw definitive conclusions on benefit, but with an at least 7 year median follow-up, the results render it less likely that major survival benefits will be achieved with this type of chemotherapy. PMID- 11378340 TI - Anti-apoptotic proteins, apoptotic and proliferative parameters and their prognostic significance in cervical carcinoma. AB - The inhibitor of apoptosis proteins (IAP) suppress apoptosis induced by a variety of stimuli. The aims of this study were to: (a) compare the expression of X linked IAP (Xiap) and Human IAP-2 (Hiap-2) in cervical carcinoma cells and normal cervix, (b) determine the correlation between IAP expression and tumour apoptosis or proliferation, and (c) assess their prognostic significance in cervical carcinomas. Paraffin-embedded tissue sections were retrieved from 77 patients with cervical squamous carcinomas prior to treatments and 47 normal subjects. Tumour apoptosis was determined by terminal deoxynucleotidyl transferase (TdT) mediated deoxyuracil triphosphate (dUTP) nick-end labelling (TUNEL) and apoptotic index (AI), and the proliferative rate was measured by Ki-67 and mitotic (MI) indices. Immunoreactive Xiap and Hiap-2 were found in both cervical cancer cells and normal tissues. IAP expressions in cancers did not correlate with apoptotic and proliferative parameters, disease stage and patient survival. The lower AI and Ki-67 index were associated with a better survival. In conclusion, the basal expression levels of IAPs have no prognostic significance, but AI and Ki-67 expression are potential prognostic indicators in cervical carcinoma. PMID- 11378341 TI - A pilot study of whole body hyperthermia and carboplatin in platinum-resistant ovarian cancer. AB - The aim of this study was to determine whether the addition of whole body hyperthermia (WBH) to carboplatin (CBDCA) can induce responses in patients with platinum-resistant ovarian cancer. 16 pretreated patients with platinum-resistant ovarian cancer were entered on a Systemic Hyperthermia Oncological Working Group (SHOWG) study; (14 patients were eligible with 14 evaluable for toxicity and 12 for response). The patients were treated with WBH (Aquatherm) 41.8 degrees C x 60 min in combination with carboplatin (CBDCA) (area under the curve (AUC) of 8) every 4 weeks. Disease status was evaluated every two cycles. Patients were treated for a maximum of six cycles. One patient had a complete response (CR) and 4 had a partial response (PR). 4 patients had stable disease (SD). 3 patients had progressive disease (PD). 2 patients were unevaluable: 1 had a bowel obstruction shortly after her first treatment; the second patient achieved a CR, but only had one treatment secondary to an idiosyncratic reaction to sedative drugs. 2 patients entered on study were ineligible, as they did not meet criteria for platinum resistance; 1 entered a CR and 1 had SD. Dose-limiting toxicity, which required CBDCA dose reductions, was grade 4 thrombocytopenia. Other toxicities included neutropenia (grade 3/4), and nausea and/or vomiting. Consistent with preclinical modelling, these results suggests that 41.8 degrees C WBH can overcome platinum resistance in ovarian cancer. These observations suggest further investigation of the therapeutic potential of WBH in a group of patients who historically fail to respond to salvage therapies is warranted. PMID- 11378342 TI - Studies on oestrogen receptor-alpha and -beta mRNA in breast cancer. AB - The oestrogen receptor (ER) is widely used to predict response to tamoxifen in patients with breast cancer. Recently a new form of ER known as ER-beta was discovered, the original ER is now designated ER-alpha. In this investigation, ER alpha and ER-beta were measured in 107 breast carcinomas and 22 fibroadenomas. Using reverse transcriptase-polymerase chain reaction (RT-PCR), ER-beta mRNA, but not ER-alpha mRNA was expressed more frequently in fibroadenomas than carcinomas. In the carcinomas, ER-beta mRNA was present in a greater proportion of samples positive for ER-alpha mRNA than in those lacking this form of the receptor. ER alpha, but not ER-beta mRNA, was significantly associated with ER protein positivity in the cancers. ER-alpha mRNA was also positively related to progesterone receptors (PR), but ER-beta mRNA showed an inverse relationship with PR. We conclude that the presently used enzyme-linked immunosorbent assay (ELISA) for ER appears to be mostly measuring ER-alpha and is unlikely to be detecting ER beta. PMID- 11378343 TI - Long-term prognosis of breast cancer patients with 10 or more positive lymph nodes treated with CMF. AB - The purpose of this investigation was to study the long-term prognosis of breast cancer patients with 10 or more positive lymph nodes after conventional chemotherapy treatment with cyclophosphamide, methotrexate and 5-fluorouracil (CMF). Between 1984 and 1989, 1048 node-positive patients were treated with CMF in two separate trials conducted by the German Breast Cancer Study Group (GBSG). Subgroups either received radiotherapy or tamoxifen in addition. In this study, long-term prognosis in the subgroup of 141 patients with 10 or more positive lymph nodes was investigated. Univariate and multivariate Cox models were used to evaluate the effect of prognostic factors on event-free survival (EFS) and overall survival (OS). Both univariate and multivariate analyses revealed the progesterone receptor (PR) status as the dominating prognostic factor for both EFS and OS, resulting in a strongly increased risk of more than 2-fold for receptor-negative patients. A large number of positive lymph nodes also affected the prognosis for EFS. In univariate analysis, the degree of lymph node involvement (i.e. percentage of positive nodes out of all examined nodes), oestrogen status (ER) status, and tumour grade also showed significant effects. To conclude, the prognosis in the subgroup of patients with 10 or more positive lymph nodes is heterogeneous. Some surprisingly high survival rates have been observed in case series of breast cancer patients treated with high-dose chemotherapy which may be explained by patient selection. From the usual factors investigated in this study, the PR status showed the strongest effect, and, at least this factor should be taken into account in the design and analysis of trials for breast cancer patients with a poor prognosis. PMID- 11378344 TI - Results of a phase III prospective, randomised trial, comparing mitoxantrone and vinorelbine (MV) in combination with standard FAC/FEC in front-line therapy of metastatic breast cancer. AB - This comparative phase III trial of mitoxantrone+vinorelbine (MV) versus 5 fluorouracil+cyclophosphamide+either doxorubicin or epirubicin (FAC/FEC) in the treatment of metastatic breast cancer was conducted to determine whether MV would produce equivalent efficacy, while resulting in an improved tolerance in relation to alopecia and nausea/vomiting. This multicentre study recruited and randomised 281 patients with metastatic breast cancer; 280 were evaluable for response survival and toxicity (138 received FAC/FEC, 142 received MV). Patient characteristics were matched in each arm and stratification for prior exposure to adjuvant therapy was made prospectively. The overall response rate (ORR) was equivalent in the two arms (33.3% for FAC/FEC versus 34.5% for MV), but MV was more effective in patients who had received prior adjuvant therapy (13% (95% confidence interval (CI) 3-23) for FAC/FEC versus 33% (95% CI 20-47) for MV P=0.025) with a better progression-free survival (PFS) (5 months (range 1-18 months) versus 8 months (range 1-27 months); P=0.0007 for FAC/FEC versus MV, respectively) while FAC/FEC was more effective in previously untreated patients (ORR 43% (95% CI 33-53) versus 35% (95% CI 25-45), P=0.26; PFS 9 months (range 0 29 months) versus 6 months (range 0-26 months) P=0.014). Toxicity was monitored through the initial six cycles of therapy; febrile neutropenia and delayed haematological recovery was more frequent for MV (P=0.001), while nausea/vomiting of grades 3-4 was greater for FAC/FEC (P=0.031), as was alopecia (P=0.0001), cardiotoxicity was the same for the two regimens. MV represents a chemotherapy combination with equivalent efficacy to standard FAC/FEC and improved results for patients who have previously received adjuvant chemotherapy. Toxicity must be balanced to allow for increased haematological suppression and risk of febrile neutropenia with MV compared with a higher risk of subjectively unpleasant side effects such as nausea/vomiting and alopecia with FAC/FEC. PMID- 11378345 TI - Veno-occlusive disease with multi-organ involvement following actinomycin-D. AB - Actinomycin-D (Act-D) is a rare cause of veno-occlusive disease (VOD). Between 1993 and 1998, we managed 6 patients, all male, median age 19 months (range 6-48 months) who received Act-D for Wilms' tumour (n=4), clear cell sarcoma (n=1) or rhabdomyosarcoma (n=1). VOD presented with a median platelet count of 12 x 10(9)/l, INR 3.8, fibrinogen 16 mg/l, fibrinogen degradation products (FDPs) > or =80 microg/l, aspartate aminotransferase (AST) 6922 IU/l, bilirubin 47 micromol/l. In 3 cases, transient liver dysfunction and thrombocytopenia without neutropenia had been observed after a previous course of Act-D. All six children developed encephalopathy, hepatomegaly, ascites, reversed portal flow and renal impairment. All received mechanical ventilation and two required haemofiltration. The treatment was supportive. Severe Adult Respiratory Distress Syndrome developed in 3 patients, all of whom died. 3 patients recovered. The outcome of VOD with multi-organ failure is poor. Intravascular coagulopathy precedes and characterises severe VOD during Act-D treatment. PMID- 11378346 TI - Incidence and prevalence of all cancerous diseases in Italy: trends and implications. AB - The burden of cancer in ageing populations is causing great concern, particularly in Italy with Europe's fastest growing elderly population. Studying all cancers combined in one group, although of limited medical value, is of great interest from the viewpoints of public health, epidemiology and the economy. Using mortality data and an estimate of cancer patients' survival we have estimated and projected incidence and prevalence in Italy of all cancers combined in one group. Five major phenomena are highlighted in the paper: (1) the decrease in the age adjusted cancer mortality rates among females and the stable mortality rates among males since 1990; (2) the changing pattern of cancer incidence since 1990, it has started to decrease for females and is stabilising for males; (3) the decrease in cancer incidence among males and females born after 1940; (4) the increase in the proportion of cancer patients that are cured with calendar years of diagnosis; (5) the increase in the total and the healthy life expectancy (i.e. cancer-free) among the Italian population since 1970. The declining and flat trends in age-adjusted cancer incidence and mortality rates since 1990 is the combined effect of survival improvements and cancer risk reduction for younger cohort groups, after 1940. These favourable trends contribute to the increase in healthy life expectation, thus supporting the idea that we live longer and healthier. PMID- 11378347 TI - Reduction in the local expression of complement component 6 (C6) and 7 (C7) mRNAs in oesophageal carcinoma. AB - Differential displays of tumour/normal pair specimens of human oesophagus identified complement component 7 (C7) as being enhanced in normal tissues, but remarkably reduced in carcinoma tissues. In situ hybridisation confirmed the localisation of C7 mRNA in normal oesophageal epithelial cells and its disappearance in tumour cells. When mRNA expressions of other components were examined by means of semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) in 10 tumour/normal pair specimens, significant reductions in C6 and C7 mRNAs were observed, while C3 and C5 mRNAs were enhanced in both normal and tumour tissues. A similar reduction was observed in colon and kidney cancers using the tumour/normal expression array analysis. Gene deletion of C7 was not found in the cell lines by Southern blot analysis. Our findings suggest a possible relationship between oesophageal tumorigenesis and reduced expression of C6 and C7 mRNAs, which is probably caused by a change in gene expression regulation and not by genetic loss of the locus. PMID- 11378348 TI - Electrochemotherapy of tumours resistant to cisplatin: a study in a murine tumour model. AB - The aim of the study was to determine whether electrochemotherapy with cisplatin could be implemented in treatment of cisplatin-resistant solid tumours. For this purpose, we used cisplatin-sensitive TBL.Cl2 cells and their cisplatin-resistant subclone TBL.Cl2 Pt, which can be grown as in vitro cell cultures and as solid subcutaneous tumours in C57Bl/6 mice. Cytotoxicity of cisplatin alone and combined with electroporation was determined by colony forming assay. Treatment effects of electrochemotherapy in vivo were assessed by tumour growth delay and tumour curability. Platinum content in the cells and tumours was determined by atomic absorption spectroscopy. In vitro, TBL.Cl2 Pt cells were equally sensitive to electrochemotherapy as their cisplatin-sensitive counterparts. In vivo, electrochemotherapy was effective on both tumour types, resulting in a prolonged tumour growth delay and tumour cures. However, electrochemotherapy was more effective on parental than cisplatin-resistant tumours, in which platinum content was significantly lower compared with parental tumours. In conclusion, electrochemotherapy is an effective treatment of cisplatin-resistant solid tumours and may prove useful in clinical chemotherapy for the treatment of tumours with intrinsic or acquired resistance to cisplatin. PMID- 11378349 TI - Different effects of methotrexate on DNA mismatch repair proficient and deficient cells. AB - Antifolates exert their antiproliferative activity through the inhibition of dihydrofolate reductase and, as a consequence, of thymidylate synthesis, thereby inducing nucleotide misincorporation and impairment of DNA synthesis. We investigated the processes involved in the repair of antifolate-induced damage and their relationship with cell death. Since misincorporated bases may be removed by DNA mismatch repair (MMR), the study was carried out on the MMR proficient human cell lines HeLa and HCT116+chr3, and, in parallel, on the MMR deficient cell lines HeLa cell-clone12, defective in the protein hPMS2, and HCT116, with an inactive hMLH1. After treatment with methotrexate (MTX), we observed that DNA repair synthesis occurs independently of the cellular MMR function. Clear signs of apoptosis such as nuclear shrinkage, chromatin condensation and degradation, DNA laddering, and poly (ADP-ribose) polymerase (PARP) proteolysis, were visible in both MMR(+) and MMR(-) cells. Remarkably, cell viability was lower and the apoptotic process was triggered more efficiently in the MMR-competent cells. PMID- 11378352 TI - Synthesis and antiviral activities of the major metabolites of the HIV protease inhibitor ABT-378 (Lopinavir). AB - The HIV protease inhibitor ABT-378 (Lopinavir) is metabolized rapidly and extensively by CYP-3A4 catalyzed oxidation. Three of the major metabolites identified were synthesized and their antiviral (HIV) activities determined. PMID- 11378353 TI - Identification of novel potent hydroxamic acid inhibitors of peptidyl deformylase and the importance of the hydroxamic acid functionality on inhibition. AB - Peptidyl deformylase (PDF) is a metallo protease that catalyzes the removal of a formyl group from the N-termini of prokaryotic prepared polypeptides, an essential step in bacterial protein synthesis. Screening of our compound collection using Staphylococcus aureus PDF afforded a very potent inhibitor with an IC(50) in the low nanomolar range. Unfortunately, the compound that contains a hydroxamic acid did not exhibit antibacterial activity (MIC). In order to address the lack of activity in the MIC assay and to determine what portion of the molecule was responsible for binding to PDF, we prepared several analogues. This paper describes our findings that the hydroxamic acid functionality found in 1 is mainly responsible for the high affinity to PDF. In addition, we identified an alternative class of PDF inhibitors, the N-hydroxy urea 18, which has both PDF and antibacterial activity. PMID- 11378354 TI - Synthesis and hydrolytic stability studies of albendazole carrier prodrugs. AB - Three N-acyl (2, 3, and 4), two N-alkoxycarbonyl (5 and 6), and one N acyloxymethyl (7) derivatives of albendazole (1) have been prepared and assessed as potential prodrugs. The determination of the aqueous solubility and partition coefficient, as well as the conversion of these derivatives to 1 in buffer solution, human plasma, and pig liver esterase were determined. PMID- 11378355 TI - A new Antennapedia-derived vector for intracellular delivery of exogenous compounds. AB - We describe the design, synthesis and cell translocation capacity of a peptide derived from the third alpha-helix of the homeodomain of Antennapedia. The new sequence appears to be an efficient and nontoxic means to deliver a covalently linked peptide cargo into cells. PMID- 11378356 TI - 6,6'-Bis(2-hydroxyphenyl)-2,2'-bipyridine manganese(III) complexes: a novel series of superoxide dismutase and catalase mimetics. AB - A series of novel manganese(III) complexes is described based on a 6,6'-bis(2 hydroxyphenyl)-2,2'-bipyridine template. These complexes show superoxide dismutase and catalase activity. The effect of the aromatic substitution pattern on the SAR is described. PMID- 11378357 TI - 5,5-Diaryl-2-amino-4-pentenoates as novel, potent, and selective glycine transporter type-2 reuptake inhibitors. AB - A novel series of 5,5-diaryl-2-amino-4-pentenoates was synthesized and found to be potent and selective glycine transporter type-2 reuptake inhibitors. PMID- 11378358 TI - Synthesis of potent and selective dopamine D(4) antagonists as candidate radioligands. AB - A series of dopamine D(4) antagonists was synthesized and evaluated as potential candidates for development as positron emission tomography (PET) radioligands. All new compounds display high affinity and selectivity for the D(4) receptors and compounds 5b, 5d, and 5e were identified as candidates for radioligand development. PMID- 11378359 TI - Structure-based design, synthesis and SAR of a novel series of thiopheneamidine urokinase plasminogen activator inhibitors. AB - The serine protease urokinase plasminogen activator (uPA) is thought to play a central role in tumor metastasis and angiogenesis. Molecular modeling studies suggest that 5-thiomethylthiopheneamidine inhibits uPA by binding at the S1 pocket of the active site. Further structure based elaboration of this residue resulted in a novel class of potent and selective inhibitors of uPA. PMID- 11378360 TI - Interaction between morphine and lysine. AB - The study by the molecular orbital theory displayed that morphine and lysine make two types of the interactions between them: type (A) by three hydrogen bondings and type (B) by one hydrogen bonding accompanied with a proton transfer. The stabilization energies were 45.3 and 34.9 kcal/mol for type (A) and type (B), respectively. The characters of these interactions are striking compared to the interactions of morphine with the other amino acids, suggesting that lysine is the binding point of morphine in the mu-opioid receptor. PMID- 11378361 TI - 4,1-Benzoxazepinone analogues of efavirenz (Sustiva) as HIV-1 reverse transcriptase inhibitors. AB - A series of 4,1-benzoxazepinone analogues of efavirenz (Sustiva) as potent NNRTIs has been discovered. The cis-3-alkylbenzoxazepinones are more potent then the trans isomers and can be synthesized preferentially by a novel stereoselective cyclization. The best compounds are potent orally bioavailable inhibitors of both wild-type HIV-1 and its clinically relevant K103N mutant virus, but are highly protein-bound in human plasma. PMID- 11378362 TI - Structure-activity relationships of phenylcyclohexene and biphenyl antitubulin compounds against plant and mammalian cells. AB - Phenylcyclohexenes (PCHs) [e.g., trans-4-nitro-5-(2,3,4 trimethoxyphenyl)cyclohexene, 2d] were found to bind weakly to the colchicine site of bovine tubulin, but are the first mimics of colchicine found to have high activity towards plant cells. Structure-activity relationships for PCHs and biphenyl AC-ring analogues of colchicine (e.g., 2,3,4,4'-tetramethoxy-2'-methyl 1,1'-biphenyl, 3e) are discussed. PMID- 11378363 TI - Synthesis and pharmacological evaluation of 2,5-cycloamino-5H-[1]benzopyrano[4,3 d]pyrimidines endowed with in vitro antiplatelet activity. AB - A series of new 2,5-cycloamino-5H-[1]benzopyrano[4,3-d]pyrimidines 3a-i have been synthesized and tested in vivo for the anti-inflammatory/analgesic/antipyretic effects and in vitro to evaluate the antiplatelet activity on guinea-pig platelet rich plasma aggregated by collagen, adenosine-5'-diphosphate (ADP) and arachidonic acid (AA). Title compounds were ineffective in vivo; however, the pyrrolidino derivatives 3a and 3c exhibited an antiplatelet activity against all the aggregants differing from that of acetylsalicylic acid (ASA) while the 5 morpholino derivatives 3g-i showed the most potent ASA-like antiplatelet activity. PMID- 11378364 TI - Indazolylamino quinazolines and pyridopyrimidines as inhibitors of the EGFr and C erbB-2. AB - Described herein is the design and synthesis of indazolylaminopyridopyrimidines and quinazolines as inhibitors of the class 1 tyrosine kinase receptor family. Data is presented for N(4)-(1-benzyl-1H-indazol-5-yl)-N(6),N(6) dimethylpyrido[3,4-d]pyrimidine-4,6-diamine 3B. This compound inhibited EGFr and c-erbB-2 enzymes selectively over other kinases. It inhibited the proliferation of a range of tumour cell lines in vitro and the growth of BT474 xenografts in SCID mice. PMID- 11378365 TI - MEK (MAPKK) inhibitors. Part 2: structure-activity relationships of 4-anilino-3 cyano-6,7-dialkoxyquinolines. AB - A series of 4-anilino-3-cyano-6,7-dialkoxyquinolines with different substituents attached to the 4-anilino group has been prepared that are potent MEK (MAP kinase kinase) inhibitors. The best activity is obtained when a phenyl or a thienyl group is attached to the para-position of the aniline through a hydrophobic linker, such as an oxygen, a sulfur, or a methylene group. The most active compounds show low nanomolar IC(50)'s against MEK (MAP kinase kinase), and have potent growth inhibitory activity in LoVo cells (human colon tumor line). PMID- 11378366 TI - Aryloxy substituted N-arylpiperazinones as dual inhibitors of farnesyltransferase and geranylgeranyltransferase-I. AB - A series of aryloxy substituted piperazinones with dual farnesyltransferase/geranylgeranyltransferase-I inhibitory activity was prepared. These compounds were found to have potent inhibitory activity in vitro and are promising agents for the inhibition of Ki-Ras signaling. PMID- 11378367 TI - Synthesis of a disulfide-linked octameric peptide construct carrying three different antigenic determinants. AB - In an effort to develop peptide vaccines against the influenza virus, we have successfully synthesized a disulfide-linked octameric homodimer that bears four copies of the influenza virus M2 protein ectodomain as well as two copies each of T-helper cell hemagglutinin epitopes, the I-E(d) restricted S1 and the I-A(d) restricted S2 fragments. Peptide attachment was via intermolecular disulfide formation from free sulfhydryl-bearing cysteine derivatives in solution. This reaction was efficient only when the amino-group of the cysteine was Fmoc protected. PMID- 11378368 TI - Modification of constrained peptides by ring-closing metathesis reaction. AB - Ring-closing metathesis (RCM) with alpha,alpha-diallylglycyl peptides is shown to furnish alpha,alpha-cyclopentenylglycyl peptides as conformationally restrained analogues in the form of post-translational type peptide modification suitable for both peptidomimetic and combinatorial chemistry applications. PMID- 11378369 TI - Effect of zinc ion on the inhibition of carboxypeptidase A by imidazole-bearing substrate analogues. AB - Competitive inhibitors of carboxypeptidase A, 2-(4-imidazoyl)hydrocinnamic acid (1) and its congeners (2-4) that bear an imidazole ring as the zinc-ligating functionality have been evaluated for their CPA inhibitory activity in the presence of zinc ion to find that the zinc ion augments the inhibitory potency by up to 212-fold. PMID- 11378370 TI - Amino acids and peptides. Part 39: a bivalent poly(ethylene glycol) hybrid containing an active site (RGD) and its synergistic site (PHSRN) of fibronectin. AB - Fibronectin contains the active sequence Arg-Gly-Asp (RGD), along with its synergic site Pro-His-Ser-Arg-Asn (PHSRN). However, the PHSRN peptide does not show synergic activity when it is mixed with the RGD peptide, indicating that a spatial array between RGD and PHSRN in fibronectin may be necessary for synergic activity. Here, we have used an amino acid type poly(ethylene glycol) derivative (aaPEG) to design a bivalent PEG hybrid of fibronectin active peptides. We prepared the aaPEG hybrid peptides PHSRN-aaPEG, aaPEG-RGD, and PHSRN-aaPEG-RGD, and tested their biological activity. Whereas aaPEG-RGD promoted cell spreading activity, PHSRN-aaPEG had no activity. The PHSRN-aaPEG-RGD hybrid strongly promoted cell spreading compared with aaPEG-RGD. These results suggest that the PHSRN sequence in the PHSRN-aaPEG-RGD molecule synergistically enhances the cell spreading activity of the RGD sequence, and that the bivalent aaPEG hybrid method may be useful for conjugating functionally active peptides. PMID- 11378371 TI - Discovery of a nuclease-resistant, non-natural dinucleotide that inhibits HIV-1 integrase. AB - Integration of HIV viral DNA into human chromosomal DNA catalyzed by HIV integrase is essential for the replication of HIV. Discovery of novel inhibitors of HIV integrase is of considerable significance in approaches to the development of therapeutic agents against AIDS. We have synthesized a new dinucleotide 1 with an internucleotide phosphate bond that is unusually resistant to exonucleases. This compound exhibits potent anti-HIV-1 integrase activity. PMID- 11378372 TI - 1,3,4-Trisubstituted pyrrolidine CCR5 receptor antagonists. Part 1: discovery of the pyrrolidine scaffold and determination of its stereochemical requirements. AB - A series of 1,3,4-trisubstituted pyrrolidines was discovered to have the ability to displace [(125)I]-MIP-1alpha from the CCR5 receptor expressed on Chinese hamster ovary (CHO) cell membranes. CCR5 activity was found to be dependent on the regiochemistry and the absolute stereochemistry of the pyrrolidine. PMID- 11378373 TI - Discovery of potent and selective phenylalanine derived CCR3 antagonists. Part 1. AB - The discovery of a series of phenylalanine derived CCR3 antagonists is reported. Parallel, solution-phase library synthesis has been utilized to delineate the structure-activity relationship leading to the synthesis of highly potent, CCR3 selective antagonists. PMID- 11378374 TI - Discovery of potent and selective phenylalanine derived CCR3 receptor antagonists. Part 2. AB - Highly potent CCR3 antagonists have been developed from a previously reported series of phenylalanine ester-based leads. Solution-phase, parallel synthesis optimization was utilized to identify highly potent, functional CCR3 antagonists. PMID- 11378375 TI - Inhibitors of the bacterial cell wall biosynthesis enzyme MurC. AB - A series of phosphinate transition-state analogues of the L-alanine adding enzyme (MurC) of bacterial peptidoglycan biosynthesis was prepared and tested as inhibitors of the Escherichia coli enzyme. Compound 4 was identified as a potent inhibitor of MurC from Escherichia coli with an IC(50) of 49nM. PMID- 11378376 TI - Hydrophobic forms of morphine-6-glucosides. AB - NMR spectroscopy of 6-acetylmorphine (6-AM), a chloroform-soluble model compound for the hydrophilic, highly potent analgesic drug morphine-6-glucoronide (M6G), in a hydrophobic solvent indicates one hydrogen bonded water molecule per molecule of 6-AM. By analysis of nuclear Overhauser enhancements (NOEs) we find a 6-AM dimer in which the monomers are linked by two water molecules. Molecular modeling studies underscore the stability of such dimeric structures involving water molecules for 6-AM and point out their more lipophilic character allowing penetration of the blood-brain barrier. PMID- 11378377 TI - Fine Tuning of physico-chemical parameters to optimise a new series of novobiocin analogues. AB - A novel series of novobiocin analogues has been synthesised by removing the lipophilic aryl chain in novobiocin and introducing an amino substituent. The structural modifications have been dictated by the control of lipophilicity and the dissociation constant of the resulting compounds. Antibacterial activity of the new coumarin derivatives could be correlated with the amount of uncharged form in physiological conditions. PMID- 11378378 TI - Arylsulphonyl hydroxamic acids: potent and selective matrix metalloproteinase inhibitors. AB - A series of novel matrix metalloproteinase inhibitors is described in which selectivity between MMP and 'sheddase' activity has been achieved and which demonstrate potent in vivo activity in models of arthritis and cancer. PMID- 11378379 TI - An aspartic protease analogue: intermolecular catalysis of peptide hydrolysis by carboxyl groups. AB - Two aspartic carboxyl groups act as key catalytic groups in the active site of an aspartic protease. We synthesized an aspartic protease analogue by positioning three salicylate residues in close proximity on a cross-linked polystyrene. The immobile artificial protease effectively hydrolyzed albumin into many small fragments by the catalytic action of carboxyl groups contained in the active site. The artificial protease manifested optimum activity at pH 3 just as aspartic proteases. PMID- 11378380 TI - Esters of 2-(1-hydroxyalkyl)-1,4-dihydroxy-9,10-anthraquinones with melphalan as multifunctional anticancer agents. AB - Eight esters of 2-(1-hydroxyalkyl)-1,4-dihydroxy-9,10-anthraquinone with melphalan were prepared and tested for their antitumor activity (S-180) and cytotoxicity. 2-[1-[4-(p-Bis(2-chloroethyl)-aminophenyl)-butanoyloxy]methyl]-1,4 dihydroxy-9,10-anthraquinone and 2-[1-[4-(p-bis(2-chloroethyl)-aminophenyl) butanoyloxy]ethyl]-1,4-dihydroxy-9,10-anthraquinone showed remarkable antitumor activity (T/C, 265 and 272%). PMID- 11378382 TI - QTL analysis identifies multiple behavioral dimensions in ethological tests of anxiety in laboratory mice. AB - BACKGROUND: Ethological tests of anxiety-related behaviors, such as the open field arena and elevated plus maze, are often carried out on transgenic animals in the attempt to correlate gene function with a behavioral phenotype. However, the interpretation of such tests is problematic, as it is probable that different tests measure different aspects of behavior; indeed, anxiety may not be a unitary phenomenon. Here, we address these questions by asking whether behaviors in five ethological tests of anxiety are under the influence of a common set of genes. RESULTS: Using over 1600 F2 intercross animals, we demonstrate that separate, but overlapping, genetic effects can be detected that influence different behavioral dimensions in the open field, elevated plus maze, square maze, light-dark box, and mirror chamber. We find quantitative trait loci (QTLs) on chromosomes 1, 4, and 15 that operate in four tests of anxiety but can be differentiated by their action on behavior in threatening and nonthreatening environments and by whether habituation of the animals to an aversive environment alters their influence. QTLs on chromosomes 7, 12, 14, 18, and X influenced a subset of behavioral measures. CONCLUSIONS: The chromosome 15 QTL acts primarily on avoidance behavior, the chromosome 1 QTL influences exploration, and the QTL on chromosome 4 influences activity. However, the effects of loci on other chromosomes are not so readily reconciled with our current understanding of the psychology of anxiety. Genetic effects on behaviors in these tests are more complex than expected and may not reflect an influence on anxiety. PMID- 11378384 TI - RNA-directed transcriptional gene silencing in plants can be inherited independently of the RNA trigger and requires Met1 for maintenance. AB - BACKGROUND: The association between DNA methylation and gene silencing has long been recognized; however, signals that initiate de novo methylation are largely unknown. In plants, recognition of RNAs that are inducers of posttranscriptional gene silencing (PTGS) can result in sequence-specific DNA methylation, and the aim of this work was to investigate whether heritable epigenetic changes can occur by this mechanism and if the Met1 methyltransferase is required. RESULTS: RNA-directed DNA methylation (RdDM) was initiated in 35S-GFP transgenic plants following infection with plant RNA viruses modified to carry portions of either the 35S promoter or the GFP coding region. Targeting of the promoter sequence resulted in both methylation and transcriptional gene silencing (TGS) that was inherited independently of the RNA trigger. Targeting the coding region also resulted in methylation; however, this was not inherited. Expression of Met1 was suppressed in order to investigate its role in initiation and maintenance of RdDM. Initiation of RdDM was found to be Met1-independent, whereas maintenance of methylation and TGS in the subsequent generations in the absence of the RNA trigger was Met1-dependent. Maintenance of methylation associated with systemic PTGS was also found to be Met1-independent. CONCLUSIONS: RNA-triggered events can lead to heritable changes in gene expression, and it is possible that initiation of other epigenetic phenomena such as trans-silencing and paramutation may have an RNA component. PMID- 11378383 TI - Completion of cytokinesis in C. elegans requires a brefeldin A-sensitive membrane accumulation at the cleavage furrow apex. AB - BACKGROUND: The terminal phase of cytokinesis in eukaryotic cells involves breakage of the intercellular canal containing the spindle midzone and resealing of the daughter cells. Recent observations suggest that the spindle midzone is required for this process. In this study, we investigated the possibility that targeted secretion in the vicinity of the spindle midzone is required for the execution of the terminal phase of cytokinesis. RESULTS: We inhibited secretion in early C. elegans embryos by treatment with brefeldin A (BFA). Using 4D recordings of dividing cells, we showed that BFA induced stereotyped failures in the terminal phase of cytokinesis; although the furrow ingressed normally, after a few minutes the furrow completely regressed, even though spindle midzone and midbody microtubules appeared normal. In addition, using an FM1-43 membrane probe, we found that membrane accumulated locally at the apices of the late cleavage furrows that form the persisting intercellular canals between daughter cells. However, in BFA-treated embryos this membrane accumulation did not occur, which possibly accounts for the observed cleavage failures. CONCLUSIONS: We have shown that BFA disrupts the terminal phase of cytokinesis in the embryonic blastomeres of C. elegans. We observed that membrane accumulates at the apices of the late cleavage furrow by means of a BFA-sensitive mechanism. We suggest that this local membrane accumulation is necessary for the completion of cytokinesis and speculate that the spindle midzone region of animal cells is functionally equivalent to the phragmoplast of plants and acts to target secretion to the equatorial plane of a cleaving cell. PMID- 11378385 TI - Hox genes and the phylogeny of the arthropods. AB - The arthropods are the most speciose, and among the most morphologically diverse, of the animal phyla. Their evolution has been the subject of intense research for well over a century, yet the relationships among the four extant arthropod subphyla - chelicerates, crustaceans, hexapods, and myriapods - are still not fully resolved. Morphological taxonomies have often placed hexapods and myriapods together (the Atelocerata) [1, 2], but recent molecular studies have generally supported a hexapod/crustacean clade [2-9]. A cluster of regulatory genes, the Hox genes, control segment identity in arthropods, and comparisons of the sequences and functions of Hox genes can reveal evolutionary relationships [10]. We used Hox gene sequences from a range of arthropod taxa, including new data from a basal hexapod and a myriapod, to estimate a phylogeny of the arthropods. Our data support the hypothesis that insects and crustaceans form a single clade within the arthropods to the exclusion of myriapods. They also suggest that myriapods are more closely allied to the chelicerates than to this insect/crustacean clade. PMID- 11378386 TI - Tumor suppressor and anti-inflammatory actions of PPARgamma agonists are mediated via upregulation of PTEN. AB - The PTEN tumor suppressor gene modulates several cellular functions, including cell migration, survival, and proliferation [1] by antagonizing phosphatidylinositol 3-kinase (PI 3-kinase)-mediated signaling cascades. Mechanisms by which the expression of PTEN is regulated are, however, unclear. The ligand-activated nuclear receptor peroxisome proliferator-activated receptor gamma (PPARgamma) [2] has been shown to regulate differentiation and/or cell growth in a number of cell types [3, 4, 5], which has led to the suggestion that PPARgamma, like PTEN [1, 6], could act as a tumor suppressor. PPARgamma has also been implicated in anti-inflammatory responses [7, 8], although downstream mediators of these effects are not well defined. Here, we show that the activation of PPARgamma by its selective ligand, rosiglitazone, upregulates PTEN expression in human macrophages, Caco2 colorectal cancer cells, and MCF7 breast cancer cells. This upregulation correlated with decreased PI 3-kinase activity as measured by reduced phosphorylation of protein kinase B. One consequence of this was that rosiglitazone treatment reduced the proliferation rate of Caco2 and MCF7 cells. Antisense-mediated disruption of PPARgamma expression prevented the upregulation of PTEN that normally accompanies monocyte differentiation and reduced the proportion of macrophages undergoing apoptosis, while electrophoretic mobility shift assays showed that PPARgamma is able to bind two response elements in the genomic sequence upstream of PTEN. Our results demonstrate a role for PPARgamma in regulating PI 3-kinase signaling by modulating PTEN expression in inflammatory and tumor-derived cells. PMID- 11378387 TI - Wnt signals are targets and mediators of Gli function. AB - There is growing evidence that Gli proteins participate in the mediation of Hedgehog and FGF signaling in neural and mesodermal development. However, little is known about which genes act downstream of Gli proteins. Here we show the regulation of members of the Wnt family by Gli proteins in different contexts. Our findings indicate that Gli2 regulates Wnt8 expression in the ventral marginal zone of the early frog embryo: activating Gli2 constructs induce ectopic Wnt8 expression in animal cap explants, whereas repressor forms inhibit its endogenous expression in the marginal zone. Using truncated Frizzled and dominant-negative Wnt constructs, we then show the requirement of at least two Wnt proteins, Wnt8 and Wnt11, for Gli2/3-induced posterior mesodermal development. Blocking Wnt signals, however, inhibits Gli2/3-induced morphogenesis, but not mesodermal specification. Gli2/3 may therefore normally coordinate the action of these two Wnt proteins, which regulate distinct downstream pathways. In addition, the finding that Gli1 consistently induces a distinct set of Wnt genes in animal cap explants and in skin tumors suggests that Wnt regulation by Gli proteins is general. Such a mechanism may link signals that induce Gli activity, such as FGFs and Hedgehogs, with Wnt function. PMID- 11378388 TI - Human populations show reduced DNA sequence variation at the factor IX locus. AB - Levels and patterns of human DNA sequence variation vary widely among loci. However, some of this variation may be due to the different populations used in different studies. So far, few studies of diverse human populations have compared different genetic loci for the same samples of populations and individuals. Here, we present new polymorphism data from intron 4 of the Factor IX gene (FIX) sequenced in diverse Old World populations. An explicit comparison is made with another X-linked gene, PDHA1, for which the sampling of individuals was very similar. Despite having a similar amount of divergence from chimpanzees, as do other nuclear genes, FIX has comparatively much less DNA sequence variation among humans. Nucleotide diversity at FIX is the lowest among the existing non-Y chromosome nuclear gene datasets and is less than 10% of the diversity found at PDHA1. Estimates of effective population size based on FIX are 8,558, about half of the value obtained for PDHA1, and the time to the most recent common ancestry among human FIX gene copies (282,000 years) is one of the most recent estimates reported for human genes. Analyses presented here suggest a history for the FIX region that includes recent positive directional selection, or background, selection. The general conclusion emerging is that very large variations can exist between the histories of similar genomic regions, even when sampling differences are minimized. PMID- 11378389 TI - A HERV-K provirus in chimpanzees, bonobos and gorillas, but not humans. AB - Evidence from DNA sequencing studies strongly indicated that humans and chimpanzees are more closely related to each other than either is to gorillas [1 4]. However, precise details of the nature of the evolutionary separation of the lineage leading to humans from those leading to the African great apes have remained uncertain. The unique insertion sites of endogenous retroviruses, like those of other transposable genetic elements, should be useful for resolving phylogenetic relationships among closely related species. We identified a human endogenous retrovirus K (HERV-K) provirus that is present at the orthologous position in the gorilla and chimpanzee genomes, but not in the human genome. Humans contain an intact preintegration site at this locus. These observations provide very strong evidence that, for some fraction of the genome, chimpanzees, bonobos, and gorillas are more closely related to each other than they are to humans. They also show that HERV-K replicated as a virus and reinfected the germline of the common ancestor of the four modern species during the period of time when the lineages were separating and demonstrate the utility of using HERV K to trace human evolution. PMID- 11378390 TI - Order of function of the budding-yeast mitotic exit-network proteins Tem1, Cdc15, Mob1, Dbf2, and Cdc5. AB - The Dbf2 protein kinase functions as part of the mitotic-exit network (MEN), which controls the inactivation of the Cdc28-Clb2 kinase in late mitosis [1]. The MEN includes the Tem1 GTP binding protein; the kinases Cdc15 and Cdc5; Mob1, a protein of unknown function; and the phosphatase Cdc14 [2]. Here we have used Dbf2 kinase activity to investigate the regulation and order of function of the MEN. We find that Tem1 acts at the top of the pathway, upstream of Cdc15, which in turn functions upstream of Mob1 and Dbf2. The Cdc5 Polo-like kinase impinges at least twice on the MEN since it negatively regulates the network, probably upstream of Tem1, and is also required again for Dbf2 kinase activation. Furthermore, we find that regulation of Dbf2 kinase activity and actin ring formation at the bud neck are causally linked. In metaphase-arrested cells, the MEN inhibitor Bub2 restrains both Dbf2 kinase activity [3] and actin ring formation [4]. We find that the MEN proteins that are required for Dbf2 kinase activity are also required for actin ring formation. Thus, the MEN is crucial for the regulation of cytokinesis, as well as mitotic exit. PMID- 11378391 TI - Transcriptional repression by suppressor of hairless involves the binding of a hairless-dCtBP complex in Drosophila. AB - Notch is the receptor for a conserved signaling pathway that regulates numerous cell fate decisions during development [1]. Signal transduction involves the presenilin-dependent intracellular processing of Notch and the nuclear translocation of the intracellular domain of Notch, NICD [2-6]. NICD associates with Suppressor of Hairless [Su(H)], a DNA binding protein, and Mastermind (Mam), a transcriptional coactivator [7-9]. In the absence of Notch signaling, Su(H) acts as a transcriptional repressor [10, 11]. Repression by Su(H) is relieved by the activation of Notch [12-16]. In the Drosophila embryo, this transcriptional switch from repression to activation is important for patterning the expression of the single-minded (sim) gene along the dorsoventral axis [12]. Here, we investigate the mechanisms by which Su(H) inhibits the expression of Notch target genes in Drosophila. We show that Hairless, an antagonist of Notch signaling [17 19], is required to repress the transcription of the sim gene. Hairless forms a DNA-bound complex with Su(H). Furthermore, it directly binds the Drosophila C terminal Binding Protein (dCtBP), which acts as a transcriptional corepressor. The dCtBP binding motif of Hairless is essential for the function of Hairless in vivo. We propose that Hairless mediates transcriptional repression by Su(H) via the recruitment of dCtBP. PMID- 11378392 TI - Fibroblast growth factor homologous factors are intracellular signaling proteins. AB - Fibroblast growth factors (FGFs) mediate cell growth, differentiation, migration, and morphogenesis by binding to the extracellular domain of cell surface receptors, triggering receptor tyrosine phosphorylation and signal transduction [1-5]. FGF homologous factors (FHFs) were discovered within vertebrate DNA sequence databases by virtue of their sequence similarity to FGFs [3, 6, 7], but the mechanism of FHF action has not been reported. We show here that FHF-1 is associated with the MAP kinase (MAPK) scaffold protein Islet-Brain-2 (IB2) [8] in the brain and in specific cell lines. FHF/IB2 interaction is highly specific, as FHFs do not bind to the related scaffold protein IB1(JIP-1b) [9, 10], nor can FGF 1 bind to IB2. We further show that FHFs enable IB2 to recruit a specific MAPK in transfected cells, and our data suggest that the scaffolds IB1 and IB2 have different MAPK specificities. Hence, FHFs are intracellular components of a tissue-specific protein kinase signaling module. PMID- 11378393 TI - Separate visual representations for perception and action revealed by saccadic eye movements. AB - Some 30 years ago, Trevarthen [1] introduced the idea of two separate visual systems, a focal system for fine motor acts and an ambient system for gross body movements such as ambulation. More recent developments indicating anatomically and physiologically separate pathways in primate vision [2] have led to a different idea of separate visual systems, one for conscious perception and one for action [3]. It has received empirical support from several studies showing that pointing, reaching, and grasping can remain accurate while the perceived position or size of objects is subject to illusory distortion [4-6]. However, much of this evidence has been challenged on the grounds of methodological flaws, particularly failure to match perfectly the conditions for verbal and motor tasks and failure to replicate results [7-10]. Here we take advantage of the strong compression of perceived position that occurs around the time of saccadic eye movements [11, 12]. Under normal lighting conditions, stimuli flashed briefly over a wide range of spatial positions just before saccadic onset are neither seen nor reached for in their veridical positions, but are compressed toward the saccadic target. We validate the idea of separate systems by showing that, in the dark, subjects are able to point accurately to the correct target position, even though their verbal reports are still subject to compression. PMID- 11378394 TI - A localized GTPase exchange factor, Bud5, determines the orientation of division axes in yeast. AB - GTPases are widespread in directing cytoskeletal rearrangements and affecting cellular organization. How they do so is not well understood. Yeast cells divide by budding, which occurs in two spatially programmed patterns, axial or bipolar [1-3]. Cytoskeletal polarization to form a bud is governed by the Ras-like GTPase, Bud1/Rsr1, in response to cortical landmarks. Bud1 is uniformly distributed on the plasma membrane, so presumably its regulators, Bud5 GTPase exchange factor and Bud2 GTPase activating protein, impart spatial specificity to Bud1 action [4]. We examined the localizations of Bud5 and Bud2. Both Bud1 regulators associate with cortical landmarks designating former division sites. In haploids, Bud5 forms double rings that encircle the mother-bud neck and split upon cytokinesis so that each progeny cell inherits Bud5 at the axial division remnant. Recruitment of Bud5 into these structures depends on known axial landmark components. In cells undergoing bipolar budding, Bud5 associates with multiple sites, in response to the bipolar landmarks. Like Bud5, Bud2 associates with the axial division remnant, but rather than being inherited, Bud2 transiently associates with the remnant in late G1, before condensing into a patch at the incipient bud site. The relative timing of Bud5 and Bud2 localizations suggests that both regulators contribute to the spatially specific control of Bud1 GTPase. PMID- 11378395 TI - Concerns mount over genetics and insurance. PMID- 11378396 TI - Fears over use of patient data for insurance. PMID- 11378397 TI - Chasing the BSE agent. PMID- 11378399 TI - Sex and death: microevolutionary trade-offs between reproductive and immune investment in dung flies. PMID- 11378398 TI - Cohesion, but not too close. PMID- 11378400 TI - Genomics: leprosy - a degenerative disease of the genome. AB - Analysis of the genome of the leprosy bacillus uncovers evidence of extensive deletion and inactivation of genes. Secluded in a specialised niche, it has discarded much of its genetic heritage, though retaining just enough to be a major human pathogen. PMID- 11378401 TI - Chromosome condensation: packaging the genome. AB - The packaging of centimetre long DNA molecules into compact metaphase chromosomes is essential for genome segregation in anaphase. The chromosomal condensin complex plays a crucial part in this packaging, and important new insight into condensin action in vitro and in vivo has recently been gained. PMID- 11378402 TI - Population genetics: the signature of selection. AB - There is hope that the structure of molecular variation within populations can give evidence for recent adaptive evolution. New work on Drosophila genes that seem to have been subject to adaptive changes illustrates the difficulties in calculating the statistical significance of data trends that seem to show this. PMID- 11378403 TI - Memory mechanisms: the yin and yang of protein phosphorylation. AB - Protein phosphorylation has long been known to play a key role in triggering the synaptic changes underlying learning and memory. Recent studies highlight the importance of tightly regulated dephosphorylation as a mechanism controlling the induction of long-term synaptic change and lasting memory. PMID- 11378404 TI - Bacterial cell division: a moving MinE sweeper boggles the MinD. AB - Placement of the division site in Escherichia coli is determined in part by three Min proteins. Recent studies have shown that MinE, previously thought to form a static ring near the division site at the midcell position, actually joins MinC and MinD in their rapid oscillation between the cell poles. PMID- 11378405 TI - Homeostasis: a scavenger receptor for haemoglobin. AB - Recent studies have identified a macrophage-specific receptor for the clearance of haemoglobin-haptoglobin complexes, which protects the host against the toxicity of free haemoglobin. PMID- 11378406 TI - Cell growth: how to grow and where to grow. AB - Root hairs provide a model system for studying tip growth in plants. The recent cloning of genes required for tip growth has shed new light on the link between ionic regulation, cell wall assembly and the cytoskeleton in cell growth. PMID- 11378407 TI - Evolutionary genetics: the evolution of plumage patterns. AB - The identification and sequencing of a gene affecting melanin production in the bananaquit, a bird species notable for its polymorphic plumage colour, paves the way for much greater understanding of the evolution of plumage patterns in birds, and the developmental modulations involved in producing new patterns. PMID- 11378408 TI - Malaria: a sporozoite runs through it. AB - A recent study reveals new insights into the development of Plasmodium sporozoites, the infectious agents of malaria. These findings may lead to changes in the approach to malaria vaccines and novel interpretations of the mechanisms of immunity to malaria. PMID- 11378409 TI - Two isoforms of a member of the arthropod defensin family from the soft tick, Ornithodoros moubata (Acari: Argasidae). AB - We previously purified and determined the partial amino acid sequence of a 4 kDa peptide having high homology with scorpion defensin from the hemolymph of adult fed female soft ticks, Ornithodoros moubata. In this study, the full length sequences of two defensin isoforms were obtained. Deduced amino acid sequences reveal a precursor protein of 73 amino acid residues with a mature portion consisting of 37 amino acid residues. This mature peptide contains six cysteine residues conserved in the same location as other invertebrate defensins. Phylogenetic analysis reveals that Ornithodoros defensin is most closely related to scorpion defensin and other more ancient arthropods. Ornithodoros defensin mRNA is constitutively expressed and up-regulated by blood-feeding and bacterial injection. Ornithodoros defensin gene expression occurs mainly in the midgut. This is the first report of the cloning and gene expression of an antibacterial peptide from the Acari. PMID- 11378410 TI - Molecular characterization of the VLDL receptor homolog mediating binding of lipophorin in oocyte of the mosquito Aedes aegypti. AB - Lipophorin (Lp) functions as a yolk protein precursor in the mosquito Aedes aegypti and it is internalized via receptor-mediated endocytosis (Insect Biochem. Mol. Biol., 30 (2000) 1161). We cloned and molecularly characterized a putative mosquito ovarian lipophorin receptor (AaLpRov) cDNA. The cDNA has a length of 3468 bp coding for a 1156-residue protein with a predicted molecular mass of 128.9 kDa. The deduced amino acid sequence of the cDNA revealed that it encodes a protein homolog of the LDL receptor superfamily, and that it harbors eight cysteine-rich ligand binding repeats at the N-terminus like vertebrate VLDL receptors. The deduced amino acid sequence of this mosquito ovarian receptor is most similar to that of the locust lipophorin receptor (LmLpR) (64.3%), and is only distantly related to the mosquito vitellogenin receptor (VgR) (18.3%), another ovarian LDLR homolog with a different ligand. The AaLpRov cDNA was expressed in a TnT Coupled Reticulocyte Lysate system, and co-immunoprecipitation experiments confirmed that the receptor protein specifically binds Lp. Developmental expression profiles clearly showed that AaLpRov transcripts are present in the vitellogenic ovary, with peak expression at 24-36 h post blood meal. In situ hybridization indicated that AaLpRov transcripts are present only in female germ line cells. Distance-based phylogenetic analyses suggest that the insect LpR and vertebrate LDL/VLDL receptor lineages separated after divergence from the insect VgR lineage. PMID- 11378411 TI - Purification and characterization of two serine protease inhibitors from the hemolymph of Mythimna unipuncta. AB - Two serine protease inhibitors, trypsin inhibitor and alpha-chymotrypsin inhibitor, were isolated from the hemolymph of Mythimna unipuncta. Mythimna trypsin and alpha-chymotrypsin inhibitors were purified by gel filtration and anion-exchange chromatography. They displayed molecular masses of 52 kDa and 43 kDa, respectively, as determined by electrophoresis under reducing and non reducing conditions on denaturing polyacrylamide gels. Their isoelectric points were evaluated by isoelectric focusing and two-dimensional electrophoresis. Their N-terminal sequences have been analyzed as APSDTTIAETLTITEEFFPD and FDESFGFQGPSTYEKTPLGEP, respectively. The role of these inhibitors in the regulation of the defense reaction of the insect is discussed. PMID- 11378412 TI - LcUSP, an ultraspiracle gene from the sheep blowfly, Lucilia cuprina: cDNA cloning, developmental expression of RNA and confirmation of function. AB - A DNA sequence corresponding to most of the DNA-binding domain of a Lucilia cuprina ultraspiracle protein (LcUSP) was amplified by PCR from genomic DNA and cloned. This cloned fragment was used to screen a L. cuprina cDNA library and to isolate a full-length LcUSP encoding sequence within a 3800-bp cDNA clone. The conceptually translated amino acid sequence of this open reading frame (467 amino acids) was used in alignment comparisons and phylogenetic analyses to reveal that LcUSP most closely resembles DmUSP relative to other known insect nuclear hormone receptors. An antisense RNA probe specific for the 5' end of Lcusp was used in ribonuclease protection assays to detect significant levels of Lcusp RNA throughout L. cuprina development. Highest levels were detected in embryos, late third instar larvae, pupae and adult females. This pattern parallels the pattern of expression observed for Dmusp RNAs during Drosophila melanogaster development. Finally, the LcUSP sequence was engineered for expression in mammalian cells and we now report that the cloned LcUSP is functional in vivo and can act as a partner for a chimeric L. cuprina ecdysone receptor to form an ecdysteroid dependent transcription factor in mammalian cells. PMID- 11378413 TI - Role of lipid transfer particle in delivery of diacylglycerol from midgut to lipophorin in larval Manduca sexta. AB - The present work analyzed the function of lipid transfer particle (LTP) in the process of exporting diacylglycerol from larval Manduca sexta midgut cells to lipophorin. When midgut sacs, which had been prelabeled in vivo with [(3)H]oleic acid, were incubated in vitro with a lipophorin-containing medium, a significant amount of radiolabeled diacylglycerol was transferred to lipophorin. Negligible amounts of diacylglycerol were released into lipophorin-free medium. In contrast, lipid-labeled lipophorin did not transfer diacylglycerol to the midgut sacs. The transfer of diacylglycerol from the midgut sac to lipophorin was blocked by preincubation of midgut sacs with antibody against LTP. Diacylglycerol transfer was restored to control values by the addition of purified LTP to midgut sacs that had been treated with antibody against LTP. Under these conditions the amount of diacylglycerol transferred was a function of the LTP concentration. These are the first results showing that LTP is required to export diacylglycerol from the midgut to lipophorin. PMID- 11378414 TI - Putative Drosophila odor receptor OR43b localizes to dendrites of olfactory neurons. AB - To gain insight into the role of the recently identified Drosophila seven transmembrane receptor family, we analyzed the cellular and subcellular localization of a member of this family, OR43b. The OR43b receptor is expressed exclusively in a subset of olfactory neurons in the third antennal segment. Consistent with a direct role in odorant transduction, receptor protein is concentrated within the dendrites, but is also present in the axons of the olfactory neurons in which it is expressed. OR43b protein is only detectable relatively late in development suggesting it may not be required for synaptic target choice of the olfactory neurons in which it is expressed. Flies carrying deletions removing one copy of OR43b have the same number of OR43b positive cells in the antenna as flies with two copies, suggesting that simple allelic exclusion of odor receptors may not occur in Drosophila. We show the OR43b gene on the balancer chromosome SM5 is expressed at reduced levels and contains nucleotide polymorphisms predicted to alter two amino acids in the receptor, including an arginine(128) to proline substitution in the first extracellular loop. The subcellular localization of OR43b in olfactory neurons supports the idea that some of the recently identified family of seven transmembrane receptors are odor receptors, and that Drosophila and vertebrates may differ in the developmental processes used to establish the neuronal architecture of the olfactory system. PMID- 11378415 TI - Stereospecificity of the (Z)-9 desaturase that converts (E)-11-tetradecenoic acid into (Z,E)-9,11-tetradecadienoic acid in the biosynthesis of Spodoptera littoralis sex pheromone. AB - Moth pheromone glands contain desaturases that catalyze the formation of conjugated dienoic fatty acids. In this article we present the first stereochemical study on one of these enzymes, namely the Delta(9) desaturase of (E)-11-tetradecenoic acid, using the moth Spodoptera littoralis as a biological model and enantiopure deuterated probes derived from tridecanoic acid. Gas chromatography coupled to mass spectrometry analysis of methanolyzed lipidic extracts from glands incubated with each individual probe showed that in the transformation of (E)-11-tetradecenoic acid into (Z,E)-9,11-tetradecadienoic acid both pro-(R) hydrogen atoms at C9 and C10 are removed from the substrate. PMID- 11378416 TI - The acetylcholinesterase gene and organophosphorus resistance in the Australian sheep blowfly, Lucilia cuprina. AB - Acetylcholinesterase (AChE), encoded by the Ace gene, is the primary target of organophosphorous (OP) and carbamate insecticides. Ace mutations have been identified in OP resistants strains of Drosophila melanogaster. However, in the Australian sheep blowfly, Lucilia cuprina, resistance in field and laboratory generated strains is determined by point mutations in the Rop-1 gene, which encodes a carboxylesterase, E3. To investigate the apparent bias for the Rop-1/E3 mechanism in the evolution of OP resistance in L. cuprina, we have cloned the Ace gene from this species and characterized its product. Southern hybridization indicates the existence of a single Ace gene in L. cuprina. The amino acid sequence of L. cuprina AChE shares 85.3% identity with D. melanogaster and 92.4% with Musca domestica AChE. Five point mutations in Ace associated with reduced sensitivity to OP insecticides have been previously detected in resistant strains of D. melanogaster. These residues are identical in susceptible strains of D. melanogaster and L. cuprina, although different codons are used. Each of the amino acid substitutions that confer OP resistance in D. melanogaster could also occur in L. cuprina by a single non-synonymous substitution. These data suggest that the resistance mechanism used in L. cuprina is determined by factors other than codon bias. The same point mutations, singly and in combination, were introduced into the Ace gene of L. cuprina by site-directed mutagenesis and the resulting AChE enzymes expressed using a baculovirus system to characterise their kinetic properties and interactions with OP insecticides. The K(m) of wild type AChE for acetylthiocholine (ASCh) is 23.13 microM and the point mutations change the affinity to the substrate. The turnover number of Lucilia AChE for ASCh was estimated to be 1.27x10(3) min(-1), similar to Drosophila or housefly AChE. The single amino acid replacements reduce the affinities of the AChE for OPs and give up to 8.7-fold OP insensitivity, while combined mutations give up to 35-fold insensitivity. However, other published studies indicate these same mutations yield higher levels of OP insensitivity in D. melanogaster and A. aegypti. The inhibition data indicate that the wild type form of AChE of L. cuprina is 12.4 fold less sensitive to OP inhibition than the susceptible form of E3, suggesting that the carboxylesterases may have a role in the protection of AChE via a sequestration mechanism. This provides a possible explanation for the bias towards the evolution of resistance via the Rop-1/E3 mechanism in L. cuprina. PMID- 11378417 TI - Characterization of two cDNAs encoding serine proteinases from the hard tick Haemaphysalis longicornis. AB - Host vaccination against tick infestation is at present the most practical and sustainable alternative tick control method to the current acaricide use which has serious limitations. However the success of this approach to control ticks depends upon the identification of target vaccine antigens. Members of the serine proteinase gene family may represent an interesting group of proteins to target as candidate antigens because of their involvement in regulation of many physiological functions and development processes in a wide range of organisms. We used RT-PCR with the 3' and 5' RACE to clone two cDNAs encoding full-length serine proteinases from the hard tick, Haemaphysalis longicornis. RT-PCR degenerate primers were designed from amino acid sequences surrounding active sites, His(57) and Ser(195) conserved among most known serine proteinase. Gene specific primers designed from nucleotide sequences of the RT-PCR products were used to prime the 3' and 5' RACE. Southern blotting analysis showed that both HLSG-1 and -2 are single copy. The 2 cDNAs, HLSG-1 and -2 are 1.2 and 1.0 kb long in size with open reading frames encoding polypeptides with 37.7 and 31.2 kDa predicted molecular mass respectively. Northern blotting analysis of total RNA from unfed and partially fed whole ticks showed that the expression of mRNAs for both HLSG-1 and -2 was induced by blood feeding. Expression analysis by RT-PCR showed that both HLSG-1 and -2 are expressed in other tick organs in addition to salivary glands and midguts. The 6 serine proteinase consensus cyteine residues are well conserved in both HLSG-1 and -2. We have discussed our findings with respect to tick vaccine development research. PMID- 11378418 TI - Isolation and developmental expression of two nuclear receptors, MHR4 and betaFTZ F1, in the tobacco hornworm, Manduca sexta. AB - The cDNAs for two members of the nuclear receptor superfamily were isolated from the tobacco hornworm, Manduca sexta. The deduced amino acid sequence of MHR4 shows 93-95% identity in the DNA-binding domain and the first portion of the hinge (D) region with the germ cell nuclear factor (GCNF)-related factors (GRFs) of the silkworm, Bombyx mori, and the mealworm, Tenebrio molitor, and with a genomic sequence from the fruit fly, Drosophila melanogaster. Northern blot hybridization showed that a 7.5 kb MHR4 mRNA appeared in Manduca abdominal epidermis just as the ecdysteroid titer began to decline during the larval molt, disappeared about 12 h later, then transiently reappeared shortly before larval ecdysis. During the pupal and adult molts, a similar pattern of expression was seen (the very end of the adult molt was not studied). At peak times of expression in the epidermis, MHR4 mRNA was also present in fat body and the central nervous system (CNS). The deduced amino acid sequence of Manduca FTZ-F1 is 100% and 96% identical to that of B. mori and Drosophila betaFTZ-F1, respectively, in the DNA-binding domain and the adjacent hinge region including the FTZ-F1 box. Northern blot analysis showed that the >9.5 kb betaFTZ-F1 mRNA appeared in Manduca epidermis during the decline of the ecdysteroid titer in the larval, pupal and adult molts as the first peak of MHR4 mRNA declined, then it disappeared in the larval and pupal molts before the second peak of MHR4 appeared. betaFTZ-F1 mRNA was also found in fat body and the CNS at the time of peak expression in the epidermis during the larval and pupal molts. Both MHR4 and betaFTZ-F1 mRNAs were found in the testis during the onset of spermatogenesis in the prepupal period. PMID- 11378419 TI - Increased variability of continuous overground walking in neuropathic patients is only indirectly related to sensory loss. AB - The present study was conducted to determine if peripheral neuropathy leads to significant changes in locomotor variability. Fourteen patients with severe peripheral neuropathy and 12 gender-, age-, height-, and weight-matched non diabetic controls participated. Sagittal plane angles of the right hip, knee, and ankle joints and tri-axial accelerations of the trunk were measured during 10 min of continuous overground walking. Standard deviations of stride times and stride to-stride standard deviations of each kinematic variable were calculated. Neuropathic patients walked slower and exhibited some increases in locomotor variability compared to control subjects. However, these increases in gait variability were primarily linked to reductions in self-selected walking speed and were not directly attributable to sensory loss itself. PMID- 11378420 TI - Changes in walking pattern caused by the possibility of a tripping reaction. AB - This study investigated in 15 young adults whether their walking pattern was altered after forewarning for a possible trip. Such changes might affect tripping reactions and consequently the validity of experimental results. Kinematics and dynamics were measured during overground walking. No changes occurred in walking velocity, step frequency, duration of stride cycle, stance, swing and double support time, or step length. A small increase was found in step width and foot clearance due to ankle dorsiflexion, but these changes were not expected to alter the probability of tripping nor the recovery reactions after tripping in an experimental setup. PMID- 11378421 TI - The effect of exercise on gait and balance in patients with chronic fatigue syndrome. AB - This study investigated anecdotal reports of gait and balance abnormalities in subjects with Chronic Fatigue Syndrome (CFS) by examining the effects of a light exercise test on postural sway and various gait parameters. Tests were performed on 11 CFS patients and 11 age- and sex-matched sedentary controls. Results demonstrated that postural sway was not significantly different in both groups before or after the exercise test. There were, however, significant differences in gait parameters between the two groups confirming anecdotal evidence, but these differences were not exacerbated by the exercise test. Heart rate responses demonstrated that both groups were exercising at similar loads, although this was perceived to be higher by the CFS group. PMID- 11378422 TI - Hemodynamics as a possible internal mechanical disturbance to balance. AB - The postural control system is assessed by observing body sway while the subject involved aims at maintaining a specified up-right posture. Internal masses generate internal reaction forces that constitute an internal mechanical stimulus that may contribute to cause segmental displacements, i.e. body sway. Thus, gaining knowledge about the amplitude and direction of these reaction forces would contribute to gain insights into the mechanisms that influence the maintenance of balance and into its control. The 3-D force vector that acts on the body centre of mass (COM) and is associated with the transient blood movement at each cardiac cycle was assessed in a population sample of 20 young adults during the maintenance of a quiet up-right posture. Typical patterns of the three components of this force vector were identified. Relevant parameters were selected and submitted to sample statistics. For a number of them, linear correlation with subject-specific parameters was found. The antero-posterior force component was characterised by a triphasic major wave, the peaks of which had values up to 0.40 N. The vertical component showed a repeatable triphasic wave with peak-to-peak values in the range 1.3-3.0 N. The medio-lateral component showed relatively low peak-to-peak values (in the range 0.05-0.10 N). The resultant vector had an amplitude that underwent several oscillations during the cardiac cycle and reached its maximal value in the range 0.6-1.7 N. PMID- 11378423 TI - Automatic postural response systems in individuals with congenital total blindness. AB - This study examined the effects of the absence of vision from birth on automatic postural responses to platform displacements during stance. Postural responses were induced by producing randomly four types of perturbations which consisted of forward and backward translations, and toe up and down rotations. Nine congenitally totally blind and nine sighted adults served as subjects. EMG signals were recorded from four muscles in the right leg, and reaction time to somatosensory stimuli generated by platform displacements was measured by pushing a hand-held button. To assess the ability to control postural balance, the root mean square (RMS) values for lateral and antero-posterior sway before, during, and after perturbations were calculated. The EMG amplitude in the gastrocnemius muscle of a blind subject was smaller than that of a sighted subject with eyes closed. No significant differences were found between blind and sighted subjects in EMG latencies of the lower extremity muscles in response to perturbations. The blind subjects had significantly faster reaction times to somatosensory stimuli triggered by platform displacements, but in toe down rotations no significant difference was found between blind and sighted subjects. The difference in the EMG latencies and reaction times between the two groups suggests that blindness from birth may not affect the spinal stretch reflex, but may affect a volitional act mediated through the motor cortex. There were also no significant differences in the RMS values for postural sway between blind and sighted subjects with eyes open or closed, although blind subjects swayed more after backward translations than did sighted subjects with eyes open. Results suggest that the ability to control postural balance during perturbations was not affected by vision loss from birth. Our findings suggest that the automatic postural response systems of humans are unaffected by the absence of vision from birth and are rather hard wired. PMID- 11378424 TI - Anticipatory postural adjustments in a bimanual, whole-body lifting task seem not only aimed at minimising anterior--posterior centre of mass displacements. AB - Anticipatory postural adjustments (APAs) were studied in a bimanual whole-body lifting task, using a mechanical analysis of the downward movement phase preceding loaded versus unloaded lifts. APAs in the backward ground reaction force were found to lead the perturbing forward box reaction with approximately 400 ms, thus inducing a backward centre of mass momentum. Both the APA onset and magnitude were scaled as a function of the load to be lifted. We conclude that, in this lifting task, the APAs served the generation of an appropriate extending moment of the ground reaction force after box pick-up, rather than the traditionally defined goal of minimising anterior-posterior centre of mass displacements. PMID- 11378425 TI - Stability analysis of four-point walking. AB - The aim of the experiment reported here was to determine the static and dynamic stability of two-point stance phases when walking on hands and knees at different speeds. In addition, we defined the methods and predicted the consequences of including two-point stance phases into crutch assisted functional electrical stimulation (FES) walking. Crawling on hands and knees was performed at three speeds by five healthy male persons. With twelve joint-position markers placed on the subject, we determined two stability indices for every instant of gait. We analysed the peak values of these two indices during the two-point stance phases. The results indicate that we have to ensure the proper position of the centre of gravity to increase the speed of walking. To reach speeds, lower than 0.6 m/s, it is not necessary to include statically unstable phases. The shift of the centre of gravity towards and across the leading stability edge can result in getting into the dynamically unstable state. Considering the results we can effectively introduce two-point stance phases into crutch assisted FES walking and therefore increase the speed and energy effectiveness of walking PMID- 11378426 TI - The evolution of clinical gait analysis part l: kinesiological EMG. AB - In 1996, I was asked by Roy Davis, President of the Gait and Clinical Movement Analysis Society, to be the presidential guest speaker at the Birmingham, AL, annual society meeting and present a talk on the development of clinical gait analysis. Following my presentation, James Gage, Editor-in-Chief for Gait and Posture, and David Winter, Associate Editor for review articles requested a manuscript for publication. To address this task I have the advantage of being a participant throughout this exciting era and of personally knowing most of the people mentioned in this manuscript. To prepare for this assignment, I wrote letters and/or made phone calls to them. Their replies to my inquiries, plus their publications, provide documentation for this review paper. The opinions expressed, for better or worse, are my own. Due to space limitations, only a partial list of the many that have contributed is presented and I regret that not all of the important contributors have been included. In some instances they will be found in Part II and Part III. Hopefully, later publications on this subject will correct the omissions. Emphasis has been given to the earliest years and to walking gait. The subject of upper extremity analysis has not been included, though studies of subjects with upper extremity motion problems are carried out in many motion laboratories including our own. A further disclaimer is that the flood of more recent publications does not receive equal coverage. History is being written daily as clinical gait analysis gains momentum. We have barely scratched the surface of the development and potential contributions of clinical gait analysis. PMID- 11378427 TI - Obesity in women of childbearing age: risks, prevention, and treatment. AB - This article reviews the health burden of obesity, its treatment and prevention, and potential barriers to care with special emphasis on adult women of childbearing age. From 1988 to 1994, 22% of nonpregnant women 18-49 years old in the United States were overweight (body mass index [BMI] >/= 25-29.9), and 22% were obese (BMI >/= 30). Both conditions increase the risk of chronic disease and mortality, and among women of childbearing age, overweight and obesity also increase the risk of infertility and adverse pregnancy outcomes.The three main strategies for preventing obesity are weight maintenance, weight loss for overweight and obese persons, and physical activity for all. More than 44% of nonpregnant women of childbearing age are trying to lose weight, and more than 33% are trying to maintain weight, but less than 21% of women of childbearing age use the recommended combination of physical activity and caloric restriction to try to lose or maintain weight. Pregnant women should try to gain no more than the recommended weight gain range for their prepregnancy BMI, yet about one third gain more weight.Although research has shown that advice from physicians can have an impact on their patients' eating habits and physical activity, many health professionals either provide no such advice or give inappropriate advice to women of childbearing age. Barriers may include inadequate reimbursement, time constraints, and lack of professional training. Frequent contact with women of childbearing age provides obstetricians and gynecologists and nurse specialists an opportunity to prevent and treat obesity successfully. PMID- 11378428 TI - Cholera. AB - Cholera, an infectious disease caused by Vibrio cholerae, is primarily transmitted by ingestion of contaminated food or water. In severe cases, cholera may lead to severe dehydration, metabolic acidosis, and ultimately, hypovolemic shock and death. The diagnosis is confirmed by identification of V. cholerae in a stool specimen. Treatment should be started immediately by rapid replacement of fluid and electrolytes. Antibiotics such as tetracycline and doxycycline shorten the duration of illness but do not significantly affect overall mortality. Cholera can be prevented by limiting spread, survival, and growth of the organism. The current parenteral cholera vaccine is not recommended by the Public Health Service or the World Health Organization because of its limited protection. A number of oral vaccines are currently being tested in clinical trials. PMID- 11378429 TI - Elevated serum CA 125 and adnexal mass mimicking ovarian carcinoma in a patient with tuberculous peritonitis. AB - A 49 year old woman presented with weight loss, diffuse abdominal pain and distension, low grade fever, massive ascites, and an increased serum CA 125 value. Computed tomography of the abdomen and pelvis and ultrasonography of the pelvis were suggestive of ovarian carcinoma. Exploratory laparotomy revealed massive ascites; diffuse peritoneal studding of numerous 1- to 2-mm implants in the bowel, omentum and abdominal wall; a cystic adnexal mass; and indurated head of the pancreas. Histopathological examination of several implants showed multiple epitheloid granulomata with rare central necrosis. The patient responded well to antituberculous therapy. PMID- 11378430 TI - The evolution of women's primary health care in california: the role of the obstetrician/gynecologist. AB - The role of the obstetrician/gynecologist as the primary health care provider for women has evolved rapidly in the past decade. There has not always been agreement in the specialty about the directions in which it should proceed. In particular, a cognitive dissonance has arisen regarding the primary care role for any one physician. Just as there are strong advocates,1,2 so are there dissentients.3,4 Access to care for women and patient choice of physician are universally acknowledged as desirable and central to the issue. The often acrimonious debate derives from more theoretical constructs of subspecialty identity and integrity, economics, dilution of specialty training, and educational barriers to successful implementation. The practical, salutary implications of primary care obstetrician/gynecologists have infrequently been evaluated in any formalized fashion. Five years ago, the State of California designated obstetrician/gynecologists as primary care physicians. This paper is a retrospective analysis of how the designation was achieved and the subsequent experience of the specialty over this period of time. PMID- 11378431 TI - Toxoplasmosis. AB - Toxoplasma gondii is a unicellular protozoan. The definitive hosts, cats, produce hardy oocysts and sporozoites. Ingestion by a nonfeline leads to the formation of tachyzoites acutely, which cause parasitemia and further dissemination, and bradyzoites, which lead to latent infection with the formation of tissue cysts in skeletal muscle, heart muscle, and central nervous system (CNS) tissue. Toxoplasmosis can be transmitted to humans by ingestion of tissue cysts in raw or inadequately cooked infected meat or in uncooked foods that have come in contact with contaminated meat, by inadvertent ingestion of oocysts and sporozoites in cat feces, or transplacentally. Immunocompetent adults and adolescents with primary infection are generally asymptomatic, but symptoms may include mild malaise, lethargy, and lymphadenopathy. Specific treatment for nonpregnant adults and adolescents is not required. Immunosuppressed patients may experience more severe manifestations, including splenomegaly, chorioretinitis, pneumonitis, encephalitis, and multisystem organ failure. These patients are also prone to reactivation of latent infection involving the CNS. All patients with human immunodeficiency virus infection and CD4 counts <100 cells per cubic millimeter should be treated prophylactically with pyrimethamine-sulfonamide. Congenital toxoplasmosis is marked by the classic triad of chorioretinits, intracranial calcifications, and hydrocephalus. Current studies have determined that prolonged treatment (1-2 years) of neonates with fansidar is important to prevent serious sequelae. Diagnosis of acute toxoplasmosis is mainly by antibody detection and generally only undertaken in pregnant patients with risk factors for transplacental transmission. All positive screening tests in pregnant women must be confirmed at a toxoplasma reference laboratory. Recent studies have shown that polymerase chain reaction testing of amniotic fluid is useful for identification or exclusion of fetal T. gondii infection. Ultrasound can be used as an adjunct to serological screening but cannot itself definitively diagnose disease. Early first-trimester maternal infections are less likely to result in congenital infection, but the sequelae are more severe. Transplacental passage is more common when maternal infection occurs in the latter half of pregnancy, but fetal injury is usually much less severe. Typically, infected pregnant patients are treated with pyrimethamine-sulfonamide for positive PCR-amniotic-fluid testing and with spiramycin for negative PCR-AF testing. PMID- 11378433 TI - Positron Emission Tomography (PET) and the Changing Face of China. PMID- 11378432 TI - Medical workplace violence. AB - Dr. Z., an obstetrician-gynecologist, refused to refill a narcotic prescription for a drug-seeking patient while on emergency room back-up call. For the next 6 months, he and his family were threatened and harassed by this patient. The constant fear of danger to his family dramatically affected Dr. Z., who questioned, "Is this the price of practicing medicine?"After months of being stalked and threatened by an angry former patient, Dr. A., an obstetrician/gynecologist, sought and was granted a court restraining order, which further infuriated the patient. When the threats became more pernicious, Dr. A. consulted a security expert, who evaluated her daily routine and advised her to dramatically revamp her life by changing where she shopped, continually altering her driving routes, learning self-defense skills, and constantly scanning her environment for the former patient. Despite making these striking changes, her life became unbearable as she loathed the practice of medicine that she once loved. Dr. A. stated, "Life will never be the same!" PMID- 11378434 TI - Combined PET/CT Imaging in Oncology. Impact on Patient Management. AB - Purpose: In this work, we describe five oncology patients whose clinical management were uniquely benefited by a novel scanner that acquires positron emission tomography (PET) and x-ray computed tomography (CT) in the same imaging session.Procedures: Co-registered 2-[F(18)]-fluoro-2-deoxy-D-glucose (FDG)-PET and CT images were acquired using a combined PET/CT scanner. Pathology and clinical follow-up data were used to confirm PET/CT scan results.Results: The combined PET/CT scanner demonstrated the ability to distinguish malignant lesions from normal physiologic FDG uptake in the striated muscles of the head and neck as well as excretory and bowel activity in the abdomen and pelvis. Additionally, the technology positively affected patient management through localization for surgical and radiation therapy planning as well as assessment of tumor response.Conclusion: Our experience indicates that simultaneous acquisition of co registered PET and CT images enabled physicians to more precisely discriminate between physiologic and malignant FDG uptake and more accurately localize lesions, improving the value of diagnostic PET in oncologic applications. PMID- 11378435 TI - Clinical Impact of 18F-FDG-PET in the Suspicion of Recurrent Ovarian Carcinoma Based on Elevated Tumor Marker Serum Levels. AB - Purpose: To retrospectively evaluate the contribution of 18F-fluorodeoxy-glucose positron emission tomography (FDG-PET) to the diagnosis and clinical management of patients who were suspected of recurrent ovarian carcinoma, based on elevated tumor markers levels with normal or equivocal computed tomography (CT) or nuclear magnetic resonance (NMR).Procedures: 20 patients with these characteristics underwent FDG-PET. PET findings were confirmed in 14, in 7 by surgery, and in the other 7 by clinical course.Results: Recurrence was confirmed in 12 patients, all with FDG-PET positive. In other 2, recurrence was rule out and in 1, FDG-PET was negative. FDG-PET accuracy was 93% with 4 surgeries avoided and guided other 6.Conclusions: FDG-PET is an useful technique for detecting recurrent ovarian carcinoma suspected by elevated tumor markers levels and normal or equivocal results in the morphologic imaging techniques and has an important clinical impact on the management of these patients. PMID- 11378436 TI - Attenuation-Correction Induced Artifact in F-18 FDG PET Imaging Following Total Knee Replacement. AB - Purpose: The clinical use of PET FDG in the work-up of patients with bone and soft tissue malignant tumors is rapidly increasing. The recognition of any source of artifact, therefore, is important to avoid interpretation pitfalls.Procedures: Two patients with complete knee joint replacement by metallic prosthesis in the course of their treatment for malignant bone and soft tissue sarcoma were evaluated by PET F-18 FDG imaging using a dual head coincidence gamma camera.Results: Both studies demonstrated in the attenuation-corrected images intense increase activity at the joint space between the metallic prosthetic surfaces at the level of the knee joint. No uptake, however, was noted in the same location on the non-attenuation-corrected images. Subsequent bone and thallium-201 scans confirmed the absence of tumor recurrence in the first patient. The second patient had multiple follow up F-18 FDG scans over a period of 16 months that show no changes from the baseline study.Conclusion: In the F-18 FDG PET images of patients with total knee metallic prosthesis, an intense activity tends to be seen in the joint space, only in the attenuation-corrected images. Such pattern of uptake is considered artifactual and should always be verified in the non-attenuated images. PMID- 11378437 TI - Cotranslational folding--omnia mea mecum porto? AB - Evidence for cotranslational folding on both prokaryotic and eukaryotic ribosomes is reviewed. Molecular chaperones appear to assist only a small fraction of newly synthesized proteins in folding into their native conformation. The recently published crystal structure of the large ribosomal subunit at 2.5 A resolution has provided the basis for understanding where and how peptide synthesis takes place on the ribosome. The nascent peptide is concluded to pass through a tunnel that extends about 100 A between the peptidyl transferase center and its exit site. The minimum diameter of the tunnel and the apparent physical and chemical properties of its walls appear to preclude complex folding of the nascent peptide within most of the length of the tunnel. However, results indicate that nascent peptides that are protected within the ribosomes vary in length from about 30 to 72 amino acid residues. This suggests that nascent peptides have different conformations. It is hypothesized that folding of the nascent polypeptide into its native conformation starts in the distal portion of the tunnel, and proceeds at the surface of the ribosomal subunit in a depression or bay near the exit opening of the tunnel. PMID- 11378438 TI - Substance p. AB - This article provides a brief overview of the history of substance P from its discovery in the 1930s to the present day. The development of substance P receptor agonists and antagonists, and more recently the employment of transgenic mice, provide a framework to explore the functional role of substance P. Chronic inflammation and pain are associated with a number of diseases, and it has been proposed that substance P, released from primary afferent nerve endings play a role in these conditions. Recent developments with substance P antagonists have demonstrated the importance of substance P in several models of disease that span from asthma to chronic bronchitis; from cystitis, inflammatory bowel disease to migraine; emesis, depression, pain and seizures. Advancements in the knowledge of the role of substance P, its agonists and antagonists could provide clinical solutions for a variety of chronic inflammatory conditions. PMID- 11378439 TI - Phosphorylation of the Fas associated factor FAF1 by protein kinase CK2 and identification of serines 289 and 291 as the in vitro phosphorylation sites. AB - We previously identified the human Fas associated factor (FAF1) as one of the interacting partners of protein kinase CK2 beta subunit. Since FAF1 is a phosphoprotein we investigated whether it is a substrate for CK2. Here, we report the full length human FAF1 cDNA sequence, expression of FAF1 in Escherichia coli and purification and characterization of FAF1 as a substrate for CK2. FAF1 as well as an N-terminal 40 kDa degradation product serve as substrates for both the recombinant CK2 holoenzyme (km 100 microM) and the isolated catalytic alpha subunit (km 200 microM). Despite the high k(m) values, we obtained evidence that CK2 is the major cellular kinase responsible for FAF1 phosphorylation, using tissue extracts as kinase sources. By MALDI-MS we identified the two serine residues at positions 289 and 291 as the major in vitro CK2 phosphorylation sites. These data may help us elucidate the functions of FAF1 and the involvement of CK2 mediated phosphorylation in processes such as apoptotic signaling, ubiquitination, nuclear translocation and embryonic development. PMID- 11378440 TI - Oxy5, a novel protein from Arabidopsis thaliana, protects mammalian cells from oxidative stress. AB - The use of molecular oxygen in various cellular processes results in the generation of toxic intracellular by-products termed reactive oxygen species (ROS). In Escherichia coli the oxyR gene product is a transcriptional regulator of the oxyR regulon that is induced in response to hydrogen peroxide-induced oxidative stress (OS). We have previously shown that an annexin-like protein from Arabidopsis thaliana, termed Oxy5, can replace the obligatory role of OxyR in E. coli. Here, we have investigated as to whether oxy5 can function across evolutionary boundaries to protect mammalian cells from OS. Overexpression of the oxyR gene in mammalian tumor cell lines protects them from hydrogen peroxide induced cell death, and these cells are also highly resistant to the superoxide ion producing compound paraquat. Oxy5 appears to be involved in the detection of calcium flux, as it binds to Ca2+ ions during hydrogen peroxide stress. Moreover, overexpression of Oxy5 leads to lowered protein kinase C activity. Thus, Oxy5 probably functions to sense and initiate protective responses to OS. In addition, Oxy5-overexpressing cells exhibit a reduction in endogenous superoxide ion levels, which concomitantly results in a dramatic decrease in their tumorigenic potential. Taken together, the results demonstrate an antioxidant role for plant oxy5 gene in mammalian cells, which can be potentially utilized in gene therapy programs aimed at reducing the deleterious effects of ROS. PMID- 11378441 TI - Low synthesis of retinoic acid due to impaired cytochrome P450 1a1 expression in mouse xeroderma pigmentosum fibroblasts. AB - New tumor formation was suppressed by retinoic acid (RA) administration in xeroderma pigmentosum (XP) patients who have a defect in nuclear excision repair. However, the inhibition is not due to enhanced removal of UV-damaged DNA. These results prompted us to investigate whether or not RA metabolism is abnormal in XP fibroblasts and what the underlying mechanism is. Compared with wild type fibroblasts, low activities of RA synthesis were determined on HPLC in mouse fibroblasts lacking XP group A (XPA) gene and UV-induced XPA deficient cancer cells. Moreover, we observed an impaired expression of cytochrome P450 1a1 in XPA deficient fibroblasts by RT-PCR and a decreased expression of retinoic acid receptor gamma in XPA deficient cancer cells by Western blotting. Finally, pre treatment of RA isoforms significantly protected the XPA deficient fibroblasts from UV-induced death. These results suggest that decreased structure activity of RA synthesis, resulting from impaired mRNA expression of cytochrome P450 1a1 may, at least together with UV irradiation, involve in skin carcinogenesis in XP patients. PMID- 11378442 TI - Effects of neurotoxin 6-aminonicotinamide on levels of enzyme activities and metabolites in quail plasma. AB - Effects of 6-aminonicotinamide (6-AN) on the levels of proteins, metabolites and enzyme activities in the plasma of Japanese quail were investigated. The concentrations of soluble proteins in the pectoral and hindlimb muscle of the 6 AN treated and the pair-fed groups were significantly reduced compared to the control group. In the plasma, the levels of total proteins and albumin were not affected, but the levels of globulin were significantly lower than those of the control and pair-fed groups. In contrast, the levels of glucose and creatine were significantly elevated. Cellulose acetate gel electrophoresis showed that 6-AN induced a new synthesis of prealbumin and also increased the levels of beta globulin relative to the control and pair-fed groups. In contrast, the levels of gamma-globulin were markedly lower than those of the control group, whereas the levels of alpha-globulin were not affected. The specific activity of alkaline phosphatase of the 6-AN group was significantly lower than that of the control and pair-fed groups and that of aspartate aminotransferase only lower than that of the control group but not the pair-fed group. The specific activities of creatine phosphokinase and lactate dehydrogenase of the 6-AN group were the greatest among the three groups, whereas those of the pair-fed group were greater than those of the control group. The results suggest that 6-AN may interfere with the proper maintenance of energy charges and the immune system function. PMID- 11378443 TI - How does hexachlorobenzene treatment affect liver uroporphyrinogen decarboxylase? AB - The aims of the present work were: (1) to investigate whether the strong decrease of liver uroporphyrinogen decarboxylase (UroD) activity observed in experimental porphyria cutanea tarda is due to alteration of the enzymatic protein and (2) to improve the knowledge about the normal liver enzyme. With these purposes, several physicochemical studies for enzymatic characterization were carried out comparatively on the 12-fold purified liver enzyme of both normal and hexachlorobenzene porphyric rat. The study shows that the enzyme from porphyric rats has a higher activation energy, lower reactivity index and lower optimum pH than the normal one. In addition, it did not reach the Vmax at any of the substrate concentrations assayed (up to 28 microM uroporphyrinogen III), while the normal enzyme reached the plateau around 14 microM. The porphyric enzyme appears to be more protected than the normal against the inhibitory action of several metals, particularly Cu2+ and Pb2+, and against thermal inactivation. Zn2+ did not affect enzymatic activity, whereas Cu2+, Hg2+, Fe2+, Pb2+, and Cd2+ lowered the activities of both normal and porphyric enzyme in a dose-related way. It was also observed that the larger the atomic radius in its hydrated state, the lower the effect of the metal. Neither glutathione nor dithiothreitol significantly altered enzymatic activity in the range of concentrations assayed. beta-Mercaptoethanol had diverse effects, as regards both the concentration assayed and the enzymatic sample used. Assays with cystine showed a dual behaviour of both normal and porphyric enzymatic activity. Western blots for both preparations revealed a single band (65 kDa) with a similar intensity. This study show that hexachlorobenzene treatment modifies the physicochemical properties of liver UroD leading to a sharp decrease of its activity, without affecting its antigenic reactivity probably as a consequence of changes at the conformational level promoted by the binding of its reported inhibitor. PMID- 11378444 TI - PDZ-binding kinase participates in spermatogenesis. AB - A cDNA clone denoted SPK (spermatogenesis-related protein kinase) was isolated from human fetal brain cDNA library. The deduced protein was 99% identical to human PDZ-binding kinase (PBK), which is a mitotic kinase. Its isoform, which is about 300 bp longer at 3' end, was also isolated in this study. SPK gene was assigned to human chromosome 8p21.2 by radiation hybrid. Northern blot analysis indicated that SPK mRNA was predominantly expressed in testis. SPK mRNA was localized to the outer cell layer of seminiferous tubules by in situ hybridization, suggesting involvement of it in the process of spermatogenesis. PMID- 11378445 TI - SECuring the perimeter. AB - Exquisitely regulated trafficking and fusion of vesicles is crucial for proper cell function. The molecules that regulate vesicle fusion are highly conserved among eukaryotes, but they have also undergone expansion and specialization within single genomes. With diversity comes the potential for functions in unique cell processes, and recent work in Arabidopsis reveals how a member of the SEC1 family, KEULE, functions in plant cell cytokinesis. PMID- 11378446 TI - Antibiotic-free chloroplast genetic engineering - an environmentally friendly approach. AB - Chloroplast genetic engineering offers several advantages over nuclear genetic engineering, including gene containment and hyperexpression. However, introducing thousands of copies of transgenes into the chloroplast genome amplifies the antibiotic resistance genes. Two recent articles report different and novel strategies to either remove antibiotic resistance genes or select chloroplast transformants without using these genes. This should eliminate their potential transfer to microorganisms or plants and ease public concerns about genetically modified crops. PMID- 11378464 TI - Suppression of gene silencing: a threat to virus-resistant transgenic plants? PMID- 11378465 TI - Is G1 arrest in plant seeds induced by a p53-related pathway? AB - In mammals, p53 is crucial for inducing the genes that lead to G1 arrest following DNA damage, enabling DNA repair. However, the possibility that such a system exists in plants has attracted little attention. Even though some plant cDNA sequences with partial homology to p53 have been reported recently, there has been little analysis of how these molecules might relate to DNA damage. The lack of investigation into whether a DNA-damage-induced, p53-mediated G1-arrest pathway might exist in plants is remarkable given that plant DNA, like that of all organisms, is continually under the threat of attack. PMID- 11378466 TI - Database-assisted promoter analysis. AB - The analysis of regulatory sequences is greatly facilitated by database-assisted bioinformatic approaches. The TRANSFAC database contains information on transcription factors and their origins, functional properties and sequence specific binding activities. Software tools enable us to screen the database with a given DNA sequence for interacting transcription factors. If a regulatory function is already attributed to this sequence then the database-assisted identification of binding sites for proteins or protein classes and subsequent experimental verification might establish functionally relevant sites within this sequence. The binding transcription factors and interacting factors might already be present in the database. PMID- 11378467 TI - How are peroxisomes formed? The role of the endoplasmic reticulum and peroxins. AB - Recent data from studies of peroxisome assembly and the subcellular sorting of peroxisomal matrix and membrane proteins have led to an expansion of the 'growth and division' and 'endoplasmic reticulum-vesiculation' models of peroxisome biogenesis into a more flexible, unified model. Within this context, we discuss the proposed role for the endoplasmic reticulum in the formation of preperoxisomes and the potential for 15 Arabidopsis peroxin homologs to function in the biogenesis of peroxisomes in plant cells. PMID- 11378468 TI - Abiotic stress signalling pathways: specificity and cross-talk. AB - Plants exhibit a variety of responses to abiotic stresses that enable them to tolerate and survive adverse conditions. As we learn more about the signalling pathways leading to these responses, it is becoming clear that they constitute a network that is interconnected at many levels. In this article, we discuss the 'cross-talk' between different signalling pathways and question whether there are any truly specific abiotic stress signalling responses. PMID- 11378469 TI - Lipoxygenase-dependent degradation of storage lipids. AB - Oilseed germination is characterized by the mobilization of storage lipids as a carbon source for the germinating seedling. In spite of the importance of lipid mobilization, its mechanism is only partially understood. Recent data suggest that a novel degradation mechanism is initiated by a 13-lipoxygenase during germination, using esterified fatty acids specifically as substrates. This 13 lipoxygenase reaction leads to a transient accumulation of ester lipid hydroperoxides in the storage lipids, and the corresponding oxygenated fatty acid moieties are preferentially removed by specific lipases. The free hydroperoxy fatty acids are subsequently reduced to their hydroxy derivatives, which might in turn undergo beta-oxidation. PMID- 11378470 TI - Aluminium tolerance in plants and the complexing role of organic acids. AB - The aluminium cation Al(3+) is toxic to many plants at micromolar concentrations. A range of plant species has evolved mechanisms that enable them to grow on acid soils where toxic concentrations of Al(3+) can limit plant growth. Organic acids play a central role in these aluminium tolerance mechanisms. Some plants detoxify aluminium in the rhizosphere by releasing organic acids that chelate aluminium. In at least two species, wheat and maize, the transport of organic acid anions out of the root cells is mediated by aluminium-activated anion channels in the plasma membrane. Other plants, including species that accumulate aluminium in their leaves, detoxify aluminium internally by forming complexes with organic acids. PMID- 11378473 TI - Genomics and proteomics converge on Helicobacter pylori. AB - During the past year, a series of studies have provided new perspectives about genetic diversity in Helicobacter pylori. The results illustrate how the current revolution in genomics and proteomics is being used to understand how this organism co-evolves with its host. The approaches should have broad applications to other host-bacterium relationships. PMID- 11378474 TI - Analysis of host responses to microbial infection using gene expression profiling. AB - Gene expression profiling offers new opportunities for understanding host-cell responses to microbial pathogens and their products. Current strategies involve either first identifying mRNAs that differ in their expression status under different experimental conditions and later defining the identity of the respective genes (for example, differential display or serial analysis of gene expression), or alternatively assessing changes in the expression of already defined genes (for example, cDNA or oligonucleotide microarrays). Early studies indicate the power of gene expression profiling for providing new insights into groups of genes whose expression is altered during the course of host-microbe interactions, and for the discovery of cellular genes that were not previously recognized to be regulated by infection. PMID- 11378475 TI - The role of gamma interferon in antimicrobial immunity. AB - Gamma interferon (IFN-gamma) is an important cytokine in the host defense against infection by viral and microbial pathogens. IFN-gamma induces a variety of physiologically significant responses that contribute to immunity. Treatment of animal cells with IFN-gamma or infection with viral or microbial pathogens leads to changes in the level of expression of several target genes as revealed by DNA microarray analyses. The signaling pathways leading to the induction of IFN-gamma regulated gene products and, in some cases, their biochemical functions have been defined in exquisite detail. Studies of transgenic mutant mice deficient in proteins of the IFN-gamma response pathway firmly establish the importance of IFN gamma in immunity. PMID- 11378476 TI - Studies of the multifaceted mast cell response to bacteria. AB - The concept of mast cells as playing a critical and multifaceted role in immune defense against pathogens is new, and effective ways to study and validate this notion are required. Recently, a number of approaches have been described that can be used to study the molecular aspects of mast cell recognition of pathogens, and of specific mast cell responses, such as mediator release, bacterial endocytosis and mast cell migration, to pathogens. PMID- 11378477 TI - Bacterial antigens elicit T cell responses via adaptive and transitional immune recognition. AB - T cells are a critical component of host immune responses against bacterial pathogens. T cell activation relies on recognition of antigen(s) derived from the bacteria, and this activation triggers potent biological effector mechanisms. Therefore, the characterization of antigens that are stimulatory for T cells provides insight into host-pathogen interactions and advances rational vaccine design. The adaptive immune response is defined by its ability to detect variable or unique single-gene products, whereas a 'transitional' immune system recognizes more conserved structures or products of multigene pathways. This transitional system functionally overlaps the canonical innate and adaptive immune responses. Antigen identification has relied upon biochemistry, genetics and expression cloning strategies. The development of computational approaches, fuelled by advances in immunology and genomic information, will facilitate the discovery of antigens and expand our understanding of both beneficial and pathological immune responses. PMID- 11378480 TI - Microbial genomes: dealing with diversity. AB - We have now complete genome sequences of several pairs of closely related prokaryotes (conspecific strains or congeneric species). Surprisingly, even strains of the same species can differ by as much as 20% in gene content. Conceptual and methodological approaches for dealing with such diversity are now being developed, and should transform microbial genomics. PMID- 11378478 TI - Host-pathogen interactions promoting inflammatory Lyme arthritis: use of mouse models for dissection of disease processes. AB - Recent studies have confirmed the infectious and inflammatory nature of arthritis induced by Borrelia burgdorferi, or Lyme arthritis. This arthritis is directed by the presence of the bacteria in joint tissue, and is mediated through activation of the Toll-like receptor 2 (TLR2) signaling pathways by borrelial lipoproteins. Several host genes regulate the severity of arthritis, possibly by regulating the balance of pro- and anti-inflammatory responses. PMID- 11378481 TI - Microbial seascapes revisited. AB - A remarkable array of new discoveries is emerging from studies of naturally occurring marine microbes. These discoveries originate from novel applications of evolving technologies, ranging from molecular phylogenetics to stable isotope analyses, to advanced microscopic techniques, to genomics. As a consequence, new perspectives on the natural history of marine microbes, the inseparable nature of the geological and biological worlds, and a plethora of unexpected new genotypes, phenotypes and physiologies are now being revealed. As our observations of naturally occurring microbes become increasingly more sophisticated, so will theory, technical applications and predictive capabilities in microbial ecology. PMID- 11378482 TI - Bacterial behavior at surfaces. AB - Population level studies demonstrate that bacterial colonization of surfaces and subsequent biofilm architecture are controlled by a variety of factors that include the hydrodynamics, surface chemistry and genotype of the cell. New molecular tools now extend our ability to investigate among bacterial cells within a surface-associated population subtle phenotypic differences that do not involve changes in genotype. Such resolution has led to new discoveries in relationships between bacterial cells and their environment. PMID- 11378483 TI - Carbon cycling: the prokaryotic contribution. AB - Although the debate continues, the concept of global warming as a consequence of the increased production of 'greenhouse gases' via human activities is now widely accepted. The role of microbes, especially the prokaryotes, in the formation, trapping and retention of 'greenhouse gases' has, for the most part, been overlooked. The future requires that we pay close attention to these organisms for possible solutions to adverse global changes. PMID- 11378484 TI - Microbial nitrogen cycles: physiology, genomics and applications. AB - Genes and pathways involved in inorganic nitrogen cycles have been found in traditional as well as unusual microorganisms. These pathways or enzymes play a very important role in the adaptation or survival of these microorganisms under a variety of environmental conditions. Microbial nitrogen metabolism has industrial applications ranging from wastewater treatment to bioremediation and potential future use in biocatalysis for chemical production. PMID- 11378485 TI - Biocides, drug resistance and microbial evolution. AB - Antimicrobial biocides are widely used in critical human health situations in which rigorous infection control is needed. Increasingly, biocidal agents are being marketed for home use, although there is little evidence that they significantly improve home hygiene. Biocide resistance mechanisms share many themes with antibiotic resistance mechanisms. PMID- 11378486 TI - Metal and radionuclide bioremediation: issues, considerations and potentials. AB - Recent demonstrations of the removal and immobilization of inorganic contaminants by microbial transformations, sorption and mineralization show the potential of both natural and engineered microbes as bioremedial tools. Demonstrations of microbe-mediated mineral formation in biofilms implicate this mode of microbial life in geological evolution and remediation of inorganic contaminants. PMID- 11378487 TI - Ethanol production from biomass: technology and commercialization status. AB - Owing to technical improvements in the processes used to produce ethanol from biomass, construction of at least two waste-to-ethanol production plants in the United States is expected to start this year. Although there are a number of robust fermentation microorganisms available, initial pretreatment of the biomass and costly cellulase enzymes remain critical targets for process and cost improvements. A highly efficient, very low-acid pretreatment process is approaching pilot testing, while research on cellulases for ethanol production is expanding at both enzyme and organism level. PMID- 11378488 TI - Microbial pathway engineering for industrial processes: evolution, combinatorial biosynthesis and rational design. AB - Microbial pathway engineering has made significant progress in multiple areas. Many examples of successful pathway engineering for specialty and fine chemicals have been reported in the past two years. Novel carotenoids and polyketides have been synthesized using molecular evolution and combinatorial strategies. In addition, rational design approaches based on metabolic control have been reported to increase metabolic flux to specific products. Experimental and computational tools have been developed to aid in design, reconstruction and analysis of non-native pathways. It is expected that a hybrid of evolutionary, combinatorial and rational design approaches will yield significant advances in the near future. PMID- 11378489 TI - Metabolic engineering as an integrating platform for strain development. AB - Integration of the analytical framework and experimental tools of metabolic engineering with emerging technologies such as DNA microarrays and directed evolution stands to dramatically improve the approaches by which strain improvement and biocatalyst design are pursued in the future. Progress in genomics and applied molecular biology, together with increasing emphasis on renewable resource utilization for chemical production, has advanced metabolic engineering to the forefront of biotechnological interest. PMID- 11378490 TI - Marine nitrogen fixation: what's the fuss? AB - Biological nitrogen fixation is a much more important process in the nitrogen cycle of the oceans than previously thought. Further, nitrogen fixation may have an influence on the capacity of the oceans to sequester carbon. A greater diversity of marine nitrogen fixers has also been uncovered but their quantitative significance remains to be determined. PMID- 11378491 TI - Rapid multiserotype detection of human rhinoviruses on optically coated silicon surfaces. AB - BACKGROUND: More than 100 immunologically distinct serotypes of human rhinoviruses (HRV) have been discovered, making detection of surface exposed capsid antigens impractical. However, the non-structural protein 3C protease (3Cpro) is essential for viral replication and is relatively highly conserved among serotypes, making it a potential target for diagnostic testing. The thin film biosensor is an assay platform that can be formatted into a sensitive immunoassay for viral proteins in clinical specimens. The technology utilizes an optically coated silicon surface to convert specific molecular binding events into visual color changes by altering the reflective properties of light through molecular thin films. OBJECTIVE: To develop a rapid test for detection of HRV by developing broadly serotype reactive antibodies to 3Cpro and utilizing them in the thin film biosensor format. STUDY DESIGN: Polyclonal antibodies to 3Cpro were purified and incorporated into the thin film assay. The in vitro sensitivity, specificity and multiserotype cross-reactivity of the 3Cpro assay were tested. Nasal washes from naturally infected individuals were also tested to verify that 3Cpro was detectable in clinical specimens. RESULTS: The 3Cpro assay is a 28-min, non-instrumented room temperature test with a visual limit of detection of 12 pM (picomolar) 3Cpro. In terms of viral titer, as few as 1000 TCID(50) equivalents of HRV2 were detectable. The assay detected 45/52 (87%) of the HRV serotypes tested but showed no cross-reactivity to common respiratory viruses or bacteria. The thin film assay detected 3Cpro in HRV-infected cell culture supernatants coincident with first appearance of cytopathic effect. Data are also presented demonstrating 3Cpro detection from clinical samples collected from HRV-infected individuals. The assay detected 3Cpro in expelled nasal secretions from a symptomatic individual on the first day of illness. In addition, 9/11 (82%) concentrated nasal wash specimens from HRV infected children were positive in the 3Cpro test. CONCLUSION: We have described a novel, sensitive thin film biosensor for rapid detection of HRV 3Cpro. This test may be suitable for the point of care setting, where rapid HRV diagnostic test results could contribute to clinical decisions regarding appropriate antibiotic or antiviral therapy. PMID- 11378492 TI - Borna disease virus--does it infect humans and cause psychiatric disorders? AB - Antibodies recognizing Borna disease virus (BDV) antigens were first demonstrated in the blood of psychiatric patients approximately 15 years ago. Since that time, a highly controversial debate arose whether BDV infects humans and whether it causes psychiatric disorders. In this review, we critically discuss the results of numerous studies that assessed this possibility by using virological and serological methods. We conclude that there is presently no strong experimental evidence supporting the notion that BDV is a human pathogen. The possibility remains, however, that an antigenically related agent is associated with human psychiatric disorders. PMID- 11378493 TI - Physical status of the E2 human papilloma virus 16 viral gene in cervical preneoplastic and neoplastic lesions. AB - BACKGROUND: Integration of human papilloma virus (HPV) 16 DNA is considered an important genetic change in cervical lesion progression towards ICC. The viral E2 gene is often disrupted by this process, releasing suppression of viral E6/E7 oncogenes, a key factor for oncogenic progression. OBJECTIVES: To evaluate the physical status of HPV 16 E2 gene in cervical preneoplastic and neoplastic lesions and its relation with lesion severity. STUDY DESIGN: A sensitive PCR approach for the detection of an intact E2 HPV 16 gene in infected epithelial cells from the cervix with low grade squamous intraepithelial lesion (LGSIL), high grade squamous intraepithelial lesion (HGSIL) and invasive cervical carcinoma (ICC) diagnosis was applied. The correlation between gene disruption and lesion stage was examined. RESULTS: Sixty-two LGSIL, 39 HGSIL and 24 ICC samples were analyzed. Fifty-seven LGSIL [92%], 13 HGSIL [33%] and 4 ICC [17%] showed results compatible with an intact E2 gene, while 5 LGSIL [8%], 26 HGSIL [67%] and 20 ICC [83%] samples gave no signal. CONCLUSIONS: HPV 16 E2 gene disruption showed a positive correlation with cervical lesion progression, particularly from LGSIL to HGSIL. Although additional genetic events are very likely to be needed for HGSIL to ICC progression, the E2 gene disruption is a putative early marker to consider in the prognostic analysis of HPV 16 chronically infected women. PMID- 11378494 TI - TT virus infection: prevalence of elevated viraemia and arguments for the immune control of viral load. AB - BACKGROUND: The most recent polymerase chain reaction (PCR) detection protocols for the TT virus (TTV) permit one to identify the presence of viral DNA in the serum of a majority of healthy individuals, in the absence of any particular risk factor. This is in contrast with previous epidemiological studies that reported a higher prevalence of TTV infection in populations such as haemodialysis patients (HD), haemophiliacs, intravenous drug users or diabetics. OBJECTIVES: To show that these discrepant results were due to the different sensitivity (number of viral copies detected) of the detection protocols used in initial and more recent epidemiological studies. STUDY DESIGN AND RESULTS: We designed a standardised primary PCR assay that detects only viraemia >5x10(3) to 5x10(4) copies/ml for genotypes 1, 2 and 3, and compared the results of this test with those of a nested PCR assay which is 100-fold more sensitive. Viraemia >5x10(3) to 5x10(4) copies/ml were statistically more frequent in HD patients (54.3%), diabetics (54.7%), and HIV-infected patients with CD4 cells <200/mm(3) (69%) than in blood donors (37%) or HIV-infected patients with CD4 cells >500/mm(3) (33%). CONCLUSIONS: These data suggest a possible relationship between the prevalence of elevated viral loads and the level of immunocompetence of the populations studied, and therefore that of an immune control of TTV viraemia. This corroborates previous findings showing that the stimulation of the immune system by an interferon treatment was able to clear TTV viraemia. PMID- 11378495 TI - Retrovirus infections in a sample of injecting drug users in Rio de Janeiro City, Brazil: prevalence of HIV-1 subtypes, and co-infection with HTLV-I/II. AB - BACKGROUND: Retrovirus infections among injecting drug users (IDUs), a core at risk population for both HIV-1 and HTLV-I/II infections in Brazil, were assessed within an ongoing cooperative research. OBJECTIVE: The study assessed the seroprevalences of HIV-1 and HTLV-I/II infections, as well as the prevalence of HIV-1 subtypes in a sample of IDUs from Rio de Janeiro, Brazil. An attempt to evaluate HIV incidence was carried out using a dual 'sensitive/less sensitive' testing strategy. STUDY DESIGN: Cross-sectional evaluation of 175 IDUs. Serostatus for HIV-1 and HTLV-I/II were established by enzyme-linked immunosorbent assays, and confirmed by western blot. The dual testing strategy aimed to estimate HIV-1 incidence rates. Differentiation between HTLV-I and -II was performed by western blot. DNA samples were polymerase chain reaction amplified by a nested protocol, and HIV-1 subtyping was determined by heteroduplex mobility assay. RESULTS: Forty-six and 29 samples were found to be, respectively, positive for HIV-1 and HTLV-I/II, 15 of them co-infected by both viruses. Among HTLV-I/II-infected patients, 75.9% were infected by HTLV-I. Thirty one HIV samples were identified as B subtype, with seven of them showing the typical "Brazilian B" pattern in the gp120 V3 loop, and ten were identified as F subtype. The use of less sensitive assays for HIV infection wrongly identified a deeply immunocompromised patient as an incident case. CONCLUSION: Moderately high seroprevalences were found for both HIV-1 and HTLV-I/II infections, HIV-1/HTLV-I co-infections being of special concern. A non-statistically significant higher prevalence of F subtype was observed, when compared with the distribution of F/B subtypes among Brazilian patients from other exposure categories. No recent HIV-1 infections were detected, but a limitation of the "sensitive/less-sensitive" testing strategy was made evident. PMID- 11378496 TI - Comparison of two commercial assays for the detection of insertion mutations of HIV-1 reverse transcriptase. AB - Insertions in the beta3-beta4 fingers subdomain of HIV-1 reverse transcriptase (RT) confer cross-resistance to various nucleoside analogs. The detection of these rearrangements in the region of codons 67-70 of RT is of primary importance for adapting and optimizing combination treatment regimen. Recent reports suggest that some genotyping techniques based on the hybridization of oligonucleotide probes may fail to detect insertion mutants of HIV-1 RT. In the present study, we have evaluated the efficiency of two commercial kits TruGene (based on Dye Primer sequencing) and Viroseq (Big Dye Terminator technique) for the detection of insertion mutations. The data were compared with an in-house dRhodamine sequencing method. Overall, all these cycle sequencing techniques were operative in the detection of insertion mutants. The best peak homogeneity in the electrophoregrams was observed with the Dye primer technique. However, specific compression artifacts were frequently encountered with this technique, rendering ambiguous the interpretation of the electrophoregrams in several regions of the sequence. This shortcoming did not occur with dRhodamine Dye terminator or Bigdye terminator cycle sequencing. In any case, a manual inspection of the electrophoregrams is highly recommended, for all types of cycle sequencing techniques, especially for detecting new mutational patterns of the RT and protease genes. Finally, some specific problems were encountered with the softwares provided with both Trugene and Viroseq kits. PMID- 11378497 TI - Cytokines and chemokines in respiratory secretion and severity of disease in infants with respiratory syncytial virus (RSV) infection. AB - BACKGROUND: little is known about inflammatory mediators (IM); like cytokines, chemokines and receptors; in respiratory secretion as possible indicators of the severity of respiratory syncytial virus (RSV) disease. Nor have systematic studies been published on the ratios between IM as such indicators. OBJECTIVE: to define the role of IM ratios as possible indicators of the severity of RSV disease. STUDY DESIGN: about 46 infants aged 0-9 months with acute RSV infections were studied. Prematurity (PM) and/or underlying disease (UD) were present in 11 of them. The concentrations of seven different IM were measured by ELISA in samples of nasopharyngeal secretions (NPS), four cytokines; IL-1, IL-6, IL-10 and TNF-alpha; the cytokine receptor TNF-R1 and the chemokines; IL-8 and RANTES. 21 IM ratios were calculated from these concentrations. The patients were assigned a clinical score (CS) ranging from 0 to 3 according to the severity of disease. RESULTS: when 25 patients with severe disease (CS 2-3) and 21 patients with mild disease (CS 0-1) were compared with respect to different IM ratios, three ratios were related to severity of disease: IL-1/RANTES, IL-8/RANTES and TNF-R1/RANTES. When 12 patients with mild disease were compared with 16 patients with severe disease, omitting patients more than 5 months of age and patients with PM and/or UD, the following IM ratios were related to severity of disease: TNF-R1/RANTES, IL-8/RANTES and RANTES/IL-10. CONCLUSION: of 21 IM ratios studied, TNF-R1/RANTES was related to severity of disease with greatest consistency. PMID- 11378498 TI - Ataxia telangiectasia: new neurons and ATM. AB - Despite the rarity of the human autosomal recessive disease ataxia telangiectasia (A-T) (affecting approximately 1/40000-1/100000), interest in the function of the mutated gene product (ATM) in this syndrome is intense. Mutation of this single gene can lead to a diverse array of features, including cancer, immune defects, infertility and radiosensitivity. However, it is the pronounced and debilitating neurodegeneration that is the hallmark of this disease. Thus, from a clinical perspective, it is ATM function in the nervous system that, arguably, is the most important to understand. Although the case for DNA damage as a causative factor for neurodegeneration in A-T is compelling, new data point to a possible link to defects in neurogenesis. Thus, whereas ATM is important for nervous system development, it could also be important for adult neurogenesis. PMID- 11378499 TI - Engineering human chromosomes for gene therapy studies. AB - We now have the capability to engineer human chromosomes that could be used to deliver therapeutic genes in gene therapy studies. These vectors have the advantages of being non-disruptive to the genome, non-immunogenic and are capable of carrying very large genes with all their regulatory sequences. What challenges lie ahead, and what future does this technology hold for gene therapy? PMID- 11378500 TI - Foci on fanconi. AB - The protein network protecting the stability of the genome is defective in Fanconi anemia (FA). The newest in a series of FA proteins is involved in DNA damage response, but the mechanism is still unclear. Clues may come from yeast two-hybrid experiments, an extraordinarily successful tool for determining molecular function. PMID- 11378513 TI - Etiology of Crohn's disease: the role of Mycobacterium avium paratuberculosis. AB - Crohn's disease is a chronic inflammatory bowel disease characterized by transmural inflammation and granuloma formation. Several theories regarding the etiology of Crohn's disease have been proposed, one of which is infection with Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis), which causes a similar disease in animals, and is present in the human food chain. Considerable evidence supports the presence of M. paratuberculosis in the intestinal tissues of many patients with Crohn's disease including culture, detection of homologous mycobacterial DNA, detection of the mycobacterial insertion sequence IS900 by both PCR and in situ hybridization in tissues, and a serologic immune response to recombinant M. paratuberculosis antigens. Despite this evidence, and our personal belief that M. paratuberculosis is a cause of Crohn's disease, widespread acceptance of this hypothesis will require evidence that specific anti-mycobacterial chemotherapy will cure the disease. PMID- 11378514 TI - Protective autoimmunity: regulation and prospects for vaccination after brain and spinal cord injuries. AB - Neuronal degeneration after traumatic injury to the central nervous system (CNS) can be reduced by active immunization or passive transfer of T cells against CNS associated myelin antigens. We propose that a protective autoimmunity is evoked by CNS insult when non-immunological local protective mechanisms cannot adequately buffer the injury-induced toxicity. The ability of a particular strain to develop a protective autoimmune response appears to be inversely related to its susceptibility to autoimmune disease. We also propose that vaccination with specific CNS-derived'safe' (non-pathogenic) peptides after traumatic CNS insult, and possibly at any stage of chronic neurodegenerative disease, can be used to boost the protective autoimmunity and thereby to reduce further injury-induced damage. Such therapeutic vaccination ensures that the augmented beneficial autoimmunity will be free of accompanying disease. PMID- 11378515 TI - Mesenchymal stem cells: building blocks for molecular medicine in the 21st century. AB - Mesenchymal stem sells (MSCs) are present in a variety of tissues during human development, and in adults they are prevalent in bone marrow. From that readily available source, MSCs can be isolated, expanded in culture, and stimulated to differentiate into bone, cartilage, muscle, marrow stroma, tendon, fat and a variety of other connective tissues. Because large numbers of MSCs can be generated in culture, tissue-engineered constructs principally composed of these cells could be re-introduced into the in vivo setting. This approach is now being explored to regenerate tissues that the body cannot naturally repair or regenerate when challenged. Moreover, MSCs can be transduced with retroviral and other vectors and are, thus, potential candidates to deliver somatic gene therapies for local or systemic pathologies. Untapped applications include both diagnostic and prognostic uses of MSCs and their descendents in healthcare management. Finally, by understanding the complex, multistep and multifactorial differentiation pathway from MSC to functional tissues, it might be possible to manipulate MSCs directly in vivo to cue the formation of elaborate, composite tissues in situ. PMID- 11378516 TI - Presenilins as therapeutic targets for the treatment of Alzheimer's disease. AB - Studies demonstrating that accumulation and aggregation of the amyloid beta protein (Abeta) within the brain is likely to cause Alzheimer's disease (AD) have provided the rationale for therapeutic strategies aimed at influencing Abeta production, aggregation and clearance. gamma-secretase catalyzes the final cleavage that releases the Abeta from its precursor; therefore, it is a potential therapeutic target for the treatment of AD. Recent data show that the polytopic membrane proteins presenilin 1 and presenilin 2 are either catalytic components or essential co-factors of a membrane-bound proteolytic complex that possesses gamma-secretase activity. Although recent findings demonstrating that gamma secretase inhibitors bind directly to presenilins (PSs) further support a catalytic role for PSs in gamma-secretase cleavage, additional studies are still needed to clarify the role of PSs in gamma-secretase cleavage and the use of targeting PSs to reduce Abeta production. PMID- 11378517 TI - Immunotherapeutic strategies for EBV-associated malignancies. AB - Advances in our understanding of the role of cytotoxic T lymphocytes (CTLs) in the control of Epstein-Barr virus (EBV)-associated malignancies and the overall biology of these diseases have led to the development of novel therapeutic strategies designed to specifically target viral antigens expressed in these malignancies. Long-term success of many of these strategies is constrained by the latency phenotypes adopted by different diseases. Adoptive transfer of polyclonal virus-specific CTLs has been used successfully to reverse the outgrowth of malignancies such as post-transplant lymphoproliferative disease (PTLD). On the other hand, limited viral gene expression in other EBV-associated malignancies such as Burkitt's lymphoma, Hodgkin's disease and nasopharyngeal carcinoma limits the efficacy of immunotherapeutic strategies used for PTLD. Preclinical studies based on specific targeting of viral antigens expressed in these malignancies have provided very encouraging results and thus are likely to serve as an important platform for the treatment of human patients. PMID- 11378518 TI - From research to implementation: challenges in the prevention of mother to child HIV transmission in the developing world. AB - Scientific research has identified anti-retroviral regimens effective in the prevention of mother to child HIV transmission. Yet, 1800 HIV-infected infants are born every day. The majority of HIV-infected women live in developing countries where challenges to widespread implementation of these regimens have prevented decreases in pediatric HIV infection from being realized. PMID- 11378519 TI - Towards mitochondrial gene therapy: DQAsomes as a strategy. AB - Mitochondrial dysfunction is a cause, or major contributing factor in the development, of degenerative diseases, aging, cancer, many cases of Alzheimer's and Parkinson's disease and Type II diabetes (D. C. Wallace, Science 283, 1482 1488, 1999). Despite major advances in understanding mtDNA defects at the genetic and biochemical level, there is no satisfactory treatment for the vast majority of patients available. Objective limitations of conventional biochemical treatment for patients with defects of mtDNA warrant the exploration of gene therapeutic approaches. However, mitochondrial gene therapy has been elusive, due to the lack of any mitochondria-specific transfection vector. We review here the current state of the development of mitochondrial DNA delivery systems. In particular, we are summarizing our own efforts in exploring the mitochondriotropic properties of dequalinium, a cationic bolaamphiphile with delocalized charge centers, for the design of a vector suited for the transport of DNA to mitochondria in living cells. PMID- 11378520 TI - Transfection by polyethyleneimine-coated microspheres. AB - Polyethyleneimine (PEI) can be used as a DNA delivery mechanism in cell culture and in vivo. Cells can be transfected by using surface-bound PEI, as well as by PEI/DNA microparticles. In the present experiments we extended these observations by preparing microspheres with covalently attached PEI. Blends of poly(epsilon CBZ-L-lysine) mixed with poly(D,L-lactic-co-glycolic acid) were formed into microspheres using a double-emulsification/solvent evaporation procedure. CBZ (carbobenzoxy) groups on the surface of microspheres were removed by Li(0) /liquid ammonia reduction. Surface amino groups were used for covalent attachment of PEI and other molecules. Silica microspheres with bonded-phase PEI were also used. Microspheres were mixed with plasmid DNA encoding green fluorescent protein and added to cultured cells. PEI-coated microspheres transfected cultured Caco cells and MH-S alveolar macrophages. Expression of the transfected DNA increased over several days. MH-S cells phagocytosed PEI-coated silica microspheres, which were shown to reside in an acidic subcellular compartment. This was demonstrated by conjugating a pH-sensitive fluorescent dye (seminaphthofluorescein, SNAFL) to the microsphere surface. Transfection of MH-S cells was increased when plasmid DNA was complexed with histone on the surface of the microspheres. CONCLUSION: PEI-coated microspheres have potential as a DNA delivery device with advantages of the unique properties of PEI and ease of surface chemical modification. PMID- 11378521 TI - Conjugation with L-Glutamate for in vivo brain drug delivery. AB - In vitro studies have shown that conjugation of a model compound [p di(hydroxyethyl)-amino-D-phenylalanine (D-MOD)] with L-Glu can improve D-MOD permeation through the bovine brain microvessel endothelial cell monolayers (Sakaeda et al., 2000). The transport of this D-MOD-L-Glu conjugate is facilitated by the L-Glu transport system. In this paper, we evaluate the in vivo brain delivery of model compounds (i.e. D-MOD, p-nitro-D-phenylalanine (p-nitro-D Phe), 5,7-dichlorokynurenic acid (DCKA) and D-kyotorphin) and their L-Glu conjugates. DCKA was also conjugated with L-Asp and L-Gln amino acids. The analgesic activities of D-kyotorphin and its L-Glu conjugate were also evaluated. The results showed that the brain-to-plasma concentration ratio of D-MOD-L-Glu was higher than the D-MOD alone; however, the plasma concentration of both compounds were the same. The plasma concentration of p-nitro-D-Phe-L-Glu conjugate was higher than the parent p-nitro-D-Phe; however, the brain-to-plasma concentration ratio of p-nitro-D-Phe was higher than its conjugate. On the other hand, both DCKA and DCKA conjugates have a low brain-to-plasma concentration ratio due to their inability to cross the blood-brain barrier (BBB). The L-Asp and L-Glu conjugates of DCKA have elevated plasma concentrations relative to DCKA; however, the DCKA-L-Gln conjugate has the same plasma concentration as DCKA. For D-kyotorphin, both the parent and the L-Glu conjugate showed similar analgesic activity. In conclusion, conjugation of a non-permeable drug with L-Glu may improve the drug's brain delivery; however, this improvement may depend on the physicochemical and receptor binding properties of the conjugate. PMID- 11378522 TI - In vivo uptake of chitosan microparticles by murine Peyer's patches: visualization studies using confocal laser scanning microscopy and immunohistochemistry. AB - Although oral vaccination has numerous advantages over parenteral injection, degradation of the vaccine and low uptake by the gut associated lymphoid tissue (GALT) still complicate the development of efficient oral vaccines. However, previous studies in our laboratory demonstrated that chitosan microparticles can have suitable size, charge, loading and release characteristics for oral vaccination using ovalbumin as model vaccine. In this study, two different approaches were used to investigate the in vivo uptake of chitosan microparticles by murine Peyer's patches. Firstly, a confocal laser scanning microscopy (CLSM) study was performed to visualize the uptake of fluorescent-labeled chitosan microparticles in the Peyer's patches after intragastrical feeding. Subsequently, the intestinal epithelial uptake of ovalbumin loaded chitosan microparticles was visualized using immunohistochemical staining of ovalbumin. Because the microparticles are biodegradable, this entrapped ovalbumin will be released after intracellular digestion in the Peyer's patches. CLSM visualization demonstrated that chitosan microparticles enhance the uptake of fluorescent-labeled ovalbumin by the epithelium of the Peyer's patches. No ovalbumin uptake by the intestinal epithelium was observed when the protein was administered without microparticles. Moreover, immunohistochemical visualization studies revealed that ovalbumin could only be transported into the Peyer's patches after association to chitosan microparticles. Since uptake by Peyer's patches is an essential step in oral vaccination, these in vivo experiments demonstrate that chitosan microparticles are very promising vaccine delivery systems. PMID- 11378523 TI - Effect of positively and negatively charged liposomes on skin permeation of drugs. AB - To clarify the effect of the surface charge of liposomes on percutaneous absorption, the permeation of liposomal drugs through rat skin was investigated in vitro and in vivo. Liposomes were prepared using egg yolk lecithin (EPC, phase transition temperature, -15 to -17 degrees C), cholesterol and dicetylphosphate (DP) or stearylamine (SA) (10:1:1, mol/mol). Also examined was the penetration behavior of positively and negatively charged liposomes, using a fluorescent probe (Nile Red). The in vitro penetration rate of melatonin (MT) entrapped in negatively charged liposomes was higher than that of positively charged ones (p<0.05). When the percutaneous absorption of ethosuximide (ES) encapsulated was estimated in vivo, the absorption of ES from negatively charged liposomes was slightly higher than that from positively charged liposomes. Additionally, the absorption of ES from both types of liposomes was superior to that from the lipid mixtures consisting of the same composition as the vesicles. The percutaneous absorption of betahistine (BH) from a gel formulation containing negatively charged liposomes of BH was much more than that from the formulation with positively charged ones, with 2-fold higher AUC (p<0.05). Histological studies revealed that the negatively charged liposomes diffused to the dermis and the lower portion of hair follicles through the stratum corneum and the follicles much faster than the positive vesicles at the initial time stage after application. Thus, the rapid penetration of negatively charged liposomes would contribute to the increased permeation of drugs through the skin. PMID- 11378524 TI - Physicochemical properties of liposomes incorporating hydrochlorothiazide and chlorothiazide. AB - In an attempt to study the effect of hydrophobic drugs on liposome properties, multilamellar liposomes (MLV) consisting of phosphatidylcholine (PC) and incorporating chlorothiazide (CT) or hydrochlorothiazide (HCT), were prepared and characterized. Liposome size, surface charge, stability (in buffer, plasma and sodium cholate) and calcium-induced aggregation were studied for drug incorporating liposomes and empty liposomes for comparison. Results show that drug incorporation affects liposome size, z-potential and stability in presence of buffer and plasma proteins. Indeed, drug-incorporating liposomes are slightly larger and have a negative surface charge, which increases with the amount of drug incorporated in the lipid membrane. The membrane integrity of drug incorporating liposomes (in absence and presence of plasma proteins) is significantly higher when compared with that of empty liposomes (for both drugs studied). On the contrary, vesicle membrane integrity in presence of sodium cholate and calcium induced vesicle aggregation, are not affected by drug incorporation. Leakage of thiazides from liposomes was demonstrated to be induced by dilution. Low amounts of thiazides (around 10-15%) are released when lipid concentration is over 0.1 mM, while further dilution increased drug leakage exponentially. Concluding, results demonstrate that the presence of HCT or CT in liposome membranes has a significant effect on main vesicle properties, which are known to influence vesicle targeting ability. Thereby, it is very interesting to continue studies in this respect, especially with more lipophilic drugs. PMID- 11378525 TI - Comparative therapeutic and toxic effects of different povidone iodine (PVP-I) formulations in a model of oral candidosis based on in vitro reconstituted epithelium. AB - In vitro models of oral candidosis based on reconstituted epithelium have been successfully used for virulence studies. In the present study we examined the effects of two povidone iodine (PVP-I) formulations (conventional PVP-I ointment, PVP-I liposome hydrogel) on reconstituted human mucosa and on a model of oral candidosis. The morphological alterations of the reconstituted mucosa caused by infection with C. albicans and by treatment were analyzed with light and electron microscopy. Specific alterations of the epithelium (vacuoles, spongiosis, oedema, detachment of keratinocytes) and invasion of the mucosa by fungal cells were reduced by treatment with the liposomal preparation, but not by the conventional ointment. However, a single application of the liposomal hydrogel to the uninfected mucosa demonstrated some structural irritations in the deeper tissue layers which were not seen with the ointment. Light microscopical studies demonstrated multiple globular structures in all samples treated with the liposomal PVP-I preparation. At the ultrastructural level these globular structures were classified as liposomes. The great majority of liposomal particles filled with dark contrasted active substance could be observed in contact with the cell wall of C. albicans and within the fungal cells. Targeting to the fungal surface resulted in a strong amount of the active ingredient next to the pathogens but not to the host cells. In conclusion the liposomal PVP-I formulation appeared superior to the conventional ointment formulation because of both less toxicity and better therapeutical effects. PMID- 11378526 TI - Commentary: possible mechanisms for strand-breaks in DNA induced by stirring. AB - When aqueous solutions of DNA are stirred at room temperature, strand breaks occur extensively. Using various spin-traps, coupled with epr spectroscopy, we have shown that this does not proceed via homolysis. It is suggested that breaks occur by hydrolysis at strongly bent regions, momentarily induced by the stirring. PMID- 11378527 TI - The iron paradox of heart and lungs and its implications for acute lung injury. AB - Iron is an essential requirement for the growth, development, and long term survival of most aerobic organisms. When control over safe iron sequestration is lost or compromised, leading to the release of low molecular mass forms of iron, the heart appears to be particularly sensitive to iron toxicity with cardiomyopathies often developing as a consequence. Iron toxicity, leading to iron-overload, is often treated in humans with the iron chelator desferrioxamine mesylate. Such treatment regimens designed to protect the heart can, however, often lead to lung injury and, in fact, several compounds with known iron chelating properties can induce severe lung dysfunction and injury. Based on these clinical observations and our recent laboratory data, we propose that the lungs actively accumulate reactive forms of iron for use in cellular growth and proliferation, and for the oxidative destruction of microbes, whereas the heart responds in the opposite way by actively removing iron which it finds extremely toxic. PMID- 11378528 TI - Quinolinic acid-iron(ii) complexes: slow autoxidation, but enhanced hydroxyl radical production in the Fenton reaction. AB - Quinolinate (pyridine-2,3-dicarboxylic acid, Quin) is a neurotoxic tryptophan metabolite produced mainly by immune-activated macrophages. It is implicated in the pathogenesis of several brain disorders including HIV-associated dementia. Previous evidence suggests that Quin may exert its neurotoxic effects not only as an agonist on the NMDA subtype of glutamate receptor, but also by a receptor independent mechanism. In this study we address ability of ferrous quinolinate chelates to generate reactive oxygen species. Autoxidation of Quin-Fe(II) complexes, followed in Hepes buffer at pH 7.4 using ferrozine as the Fe(II) detector, was found to be markedly slower in comparison with iron unchelated or complexed to citrate or ADP. The rate of Quin-Fe(II) autoxidation depends on pH (squared hydroxide anion concentration), is catalyzed by inorganic phosphate, and in both Hepes and phosphate buffers inversely depends on Quin concentration. These observations can be explained in terms of anion catalysis of hexaaquairon(II) autoxidation, acting mainly on the unchelated or partially chelated pool of iron. In order to follow hydroxyl radical generation in the Fenton chemistry, electron paramagnetic resonance (EPR) spin trapping with 5,5 dimethyl-1-pyrroline-N-oxide (DMPO) was employed. In the mixture consisting of 100 mM DMPO, 0.1 mM Fe(II), and 8.8 mM hydrogen peroxide in phosphate buffer pH 7.4, 0.5 mM Quin approximately doubled the yield of DMPO-OH adduct, and higher Quin concentration increased the spin adduct signal even more. When DMPO-OH was pre-formed using Ti3+ /hydrogen peroxide followed by peroxide removal with catalase, only addition of Quin-Fe(II), but not Fe(II), Fe(III), or Quin-Fe(III), significantly promoted decomposition of pre-formed DMPO-OH. Furthermore, reaction of Quin-Fe(II) with hydrogen peroxide leads to initial iron oxidation followed by appearance of iron redox cycling, detected as slow accumulation of ferrous ferrozine complex. This phenomenon cannot be abolished by subsequent addition of catalase. Thus, we propose that redox cycling of iron by a Quin derivative, formed by initial attack of hydroxyl radicals on Quin, rather than effects of iron complexes on DMPO-OH stability or redox cycling by hydrogen peroxide, is responsible for enhanced DMPO-OH signal in the presence of Quin. The present observations suggest that Quin-Fe(II) complexes display significant pro-oxidant characteristics that could have implications for Quin neurotoxicity. PMID- 11378529 TI - The inhibition of oxygen radical release from human neutrophils by resting platelets is reversed by administration of acetylsalicylic acid or clopidogrel. AB - Resting platelets inhibit oxygen radical release from neutrophils. Antiplatelet therapy may support this function by preventing platelet activation. Whether antiplatelet agents affect the antioxidative action of resting platelets in the absence of platelet activation is unknown. The effect of acetylsalicylic acid or clopidogrel administration on the antioxidative action of resting platelets was therefore studied in ten healthy volunteers. Preparations of resting platelets were obtained from 5 subjects each - before, during and after an eight-day course of daily treatment with 100 mg of acetylsalicylic acid or 75 mg of the thienopyridine clopidogrel. Human peripheral blood neutrophils were pretreated with the platelets at a ratio of (1/5)0 for 45 min; then formyl-Met-Leu-Phe triggered oxygen radical release was measured fluorometrically. The inhibitory effect of platelets on oxygen radical release from neutrophils which was seen before treatment was abolished by antiplatelet therapy with either of the drugs, and inhibition was restored gradually after discontinuing acetlsalicylic acid/ clopidogrel intake. Results suggest that the protective role of resting platelets in controlling oxygen radical release from neutrophils in the absence of platelet activation may be impaired by antiplatelet therapy. PMID- 11378530 TI - Peroxynitrite decay in the presence of hydrogen peroxide, mannitol and ethanol: a reappraisal. AB - We have reported previously that the apparent rate of peroxynitrite (ONOO(-) ) decay, as followed from its absorbance at 302 nm, decreases in the presence of hydrogen peroxide, mannitol and ethanol (Alvarez et al., 1995, Chem. Res. Toxicol. 8:859-864; Alvarez et al., 1998, Free Radic. Biol. Med. 24:1331-1337). Recently, two papers confirmed the observation and proposed that this slowing effect was due to the formation of absorbing peroxynitrate (O(2) NOO(-) ) as intermediate (Goldstein and Czapski, 1998, J. Am. Chem. Soc. 120:3458-3463; Hodges and Ingold, 1999, J. Am. Chem. Soc. 121:10695-10701). Peroxynitrate would be formed from the reaction of peroxynitrite-derived nitrogen dioxide with superoxide. Superoxide, in turn, would arise from the one-electron oxidation of hydrogen peroxide, or from the reaction of reductive radicals derived from mannitol and ethanol with dioxygen. In agreement with this concept, we show herein that under the conditions of our previous work, the slowing effect is prevented by superoxide dismutase and, in the case of mannitol and ethanol, by reducing the dioxygen concentration of the reaction solutions. Thus, superoxide formation is necessary for the decrease in the rate of absorbance decay. In addition, by simulations using known rate constants and absorption coefficients, we show that the slowing effect can be quantitatively accounted for by the formation of peroxynitrate. PMID- 11378531 TI - Lipid peroxidation is increased in paraoxonase L55 homozygotes compared with M allele carriers. AB - Human serum paraoxonase (PON) is an antioxidative enzyme, which circulates on high-density lipoproteins and appears to use oxidized phospholipids as physiological substrates. PON M/L55 substitution changes the ability of PON to prevent lipid oxidation. Urinary 8-iso-PGF(2alpha) (one of F2 -isoprostanes) may represent a non-invasive in vivo index of free radical generation and we propose that PON might influence the biosynthesis of 8-iso-PGF(2alpha) in the vasculature. We studied the urinary excretion of 8-iso-PGF(2alpha) and related it to PON M/L55 genotypes in patients with type 2 diabetes mellitus (n = 55) and non diabetic control subjects (n = 55). Urinary 8-iso-PGF(2alpha) was determined by competitive ELISA and the PON genotype by a PCR based restriction enzyme digestion method. LL homozygotes were compared to M-allele carriers (ML heterozygotes and MM homozygotes). The urinary excretion of 8-iso-PGF(2alpha) among non-diabetic non-smoking LL homozygotes was 3995.5 +/- 3352.8 ng/24-hour and among M-allele carriers 1689.8 +/- 1051.3 ng/24-hour (p = 0.017, ANCOVA; gender, hypertension, total cholesterol, triglycerides and LDL cholesterol as covariates). The excretion of 8-iso-PGF(2alpha), was increased in type 2 diabetes mellitus compared to non-diabetic control subjects. PON may thus protect against oxidative stress by destroying some biologically active lipids. Excretion of 8 iso-PGF(2alpha) is increased in type 2 diabetes, which may reflect oxidant injury. PMID- 11378532 TI - Selective distribution of oxysterols in atherosclerotic lesions and human plasma lipoproteins. AB - The presence of oxidized sterols (oxysterols) in human serum and lesions has been linked to the initiation and progression of atherosclerosis. Data concerning the origin, identity and quantity of oxysterols in biological samples are controversial and inconsistent. This inconsistency may arise from different analytical methods or handling conditions used by different investigators. In the present study, oxysterol levels and distribution were analyzed by an optimized GC MS method, in human atherosclerotic coronary and carotid lesions, in atherosclerotic apolipoprotein E deficient mice (E degrees mice) and in native and in vitro oxidized human low and high density lipoproteins. Oxysterol levels were analyzed with a limit of detection of 0.06 - 0.24 ng, with 25 hydroxycholesterol (25-OH) being the least sensitive. In human coronary and carotid lesions, obtained from endatherectomic samples, 27-hydroxycholesterol (27 OH) was the major oxysterol, with about 85% as sterols esterified to fatty acids. While total cholesterol and oxysterols levels were similar in both kinds of human lesions, oxysterol distribution was significantly different. In coronary lesions the mean levels of 27-OH and 7beta-hydroxycholesterol (7beta-OH) were 38% and 20% of total oxysterols, whereas in carotid lesions their mean levels were 66% and 5%, respectively. Unlike in human aortic lesions, 27-OH was entirely absent in E degrees mice, whereas the level of 7alpha-hydroxycholesterol (7alpha-OH) was 28% of the total oxysterols, vs. 5% in human coronary lesions. As 27-OH is an enzymatic product of cholesterol oxidation, this finding may indicate that such an enzymatic process does not take place in E degrees mice. PMID- 11378533 TI - Effects of hypochlorite on cultured respiratory epithelial cells. AB - Neutrophils and eosinophils are involved in the pathogenesis of many respiratory diseases. The enzymes myeloperoxidase and eosinophil peroxidase catalyze the reaction of H2 O2 with Cl to produce the reactive oxygen species HOCl. Normal human bronchial epithelial (NHBE) cells were exposed to 0.18-0.90 mM HOCl for 48 h, and studied with immunohistochemical, metabolic and morphological studies. The ability of the cells to attach to each other and/or to the matrix was altered. Immunohistochemical studies showed a decreased amount of desmosomes and focal adhesion sites, although the morphology of the cells was not affected. The ability of the mitochondria to oxidize glucose was reduced. HOCl-exposed cells had an increased production of NO, probably by an increased activity of cNOS, due to increased intracellular Ca2+. The antioxidant N-acetylcysteine inhibited both the NO production and the effects of HOCl on glucose oxidation. The cNOS inhibitor N-propyl-L-arginine inhibited HOCl-induced NO production. X-ray microanalysis showed an increase in the intracellular Na+ /K+ ratio, which indicates cell damage. In conclusion, exposure to HOCl results in cell detachment and metabolic alterations in normal human bronchial epithelial cells. Oxygen radicals could in part mediate the effects. Oxygen radicals could hence contribute to the observed epithelial damage in respiratory diseases. PMID- 11378534 TI - Peroxynitrite promotes mitochondrial permeability transition-dependent rapid U937 cell necrosis: survivors proliferate with kinetics superimposable on those of untreated cells. AB - A short term exposure to peroxynitrite promotes a time- and concentration dependent lethal response in U937 cells. The mode of cell death was necrosis and rapid (within minutes) cell lysis was found to occur via a mechanism involving mitochondrial permeability transition. Apoptosis was not detected in cells exposed to low levels of peroxynitrite, or in cells which survived a treatment with toxic amounts of peroxynitrite, neither after the 60 min exposure nor following increasing time intervals of growth in fresh culture medium. Rather, cells treated with peroxynitrite concentrations which were not immediately lethal, as well as the survivors of treatments with toxic levels of peroxynitrite, proliferated with kinetics superimposable on those observed in untreated cells. PMID- 11378535 TI - Oral supplements of vitamin E improve measures of oxidative stress in plasma and reduce oxidative damage to LDL and erythrocytes in beta-thalassemia intermedia patients. AB - Fifteen beta-thalassemia intermedia patients, not requiring chronic transfusional therapy, were monitored in order to check their antioxidant status, and the lipid oxidation products in plasma, LDL, and erythrocytes before and during a 9-month oral treatment with 600 mg/day vitamin E. The low level of vitamin E, and high level of malondialdehyde in plasma clearly tended to normalize after three months (P < .001), and were quite similar to control after six months. The abnormally low level of vitamin E in LDL and the four times higher than control basal level of conjugated dienes (LDL-CD), were not modified after three months of treatment. Significant changes of LDL-VE (P < .05) and of the basal LDL-CD (P < .001) were evident after six months. LDL-VE was within the normal range after nine months, whereas LDL-CD still appeared twice as higher than control. Plasma vitamin A, ascorbate, beta-carotene, and lycopene increased markedly at the end of the trial (P < .005). The level of vitamin E in red blood cells was normalized after six months of supplementation. A decrease of the baseline value of conjugated dienes was observed after nine months, although it remained 1.4-fold higher than control. The RBC count and hematocrit appeared higher at the end of the trial (P < .05 and P < .001, respectively). The hemoglobin value did not show variations. A shift to normal of the resistance of erythrocytes to osmotic lysis was observed. Our findings provide evidence that an oral treatment with vitamin E improves the antioxidant/oxidant balance in plasma, LDL particles, and red blood cells, and counteracts lipid peroxidation processes in beta-thalassemia intermedia patients. PMID- 11378536 TI - Vascular endothelial growth factor/vascular permeability factor in the pathogenesis of primary effusion lymphomas. AB - Primary effusion lymphomas (PEL), rare lymphomas associated with Kaposi's sarcoma associated herpesvirus (KSHV or HHV-8) infection, present as malignant lymphomatous effusions in body cavities. We have recently found that PEL effusions contain high levels of vascular endothelial growth factor/vascular permeability factor (VEGF/VPF). VEGF/VPF, an important regulator of tumor angiogenesis in vivo, exerts its effects acting through the receptors KDR/Flk-1 and Flt-1 on the endothelial cell membrane. In vitro, the PEL cell lines BC-1, BCP-1 and BCBL-1 produce high levels of VEGF. RT-PCR analysis of RNA from the PEL cell lines amplified the three VEGF/VPF secreted isoforms, VEGF/VPF(121), VEGF/VPF(145) and VEGF/VPF(165). Two of the PEL cell lines express the VEGF/VPF receptor Flt-1, but VEGF did not stimulate proliferation in these cells. SCID/beige mice inoculated intraperitoneally with BCBL-1 cells developed effusion lymphoma of human cell origin with prominent bloody ascites. In contrast, none of the mice treated with a neutralizing anti-human VEGF/VPF antibody developed ascites and effusion lymphoma. Although the precise mechanisms by which VEGF/VPF can promote vascular permeability are not fully understood, VEGF/VPF stimulation of vascular leakage may be critical to the pathogenesis of PEL. PMID- 11378537 TI - VLA-4-dependent myeloma cell adhesion. AB - In multiple myeloma, the malignant plasma cells in the bone marrow attach to stromal elements using several adhesion receptors. The integrin VLA-4 plays a major role in mediating myeloma cell adhesion to the stroma, by interacting with its ligands VCAM-1 and fibronectin. VLA-4, as well as other integrins, can be expressed in different states of activation, which convey different levels of adhesion. Modulation of VLA-4 adhesive activity by external stimuli has been demonstrated for hematopoietic progenitor cells and lymphocytes, and can represent a potential mechanism contributing to the localization of myeloma cells in the bone marrow. In this review we have summarized data on the characterization of VLA-4-mediated myeloma cell adhesion, and we present potential mechanisms of modulation of VLA-4 activity. In addition, we also speculate on the signalling generated upon interaction of myeloma VLA-4 with its ligands on the survival of these cells in the bone marrow. PMID- 11378538 TI - Caspases: conductors of the cell death machinery in lymphoma cells. AB - The present review focuses on recent insights into the regulation of caspases by other components of the apoptotic pathway, including the mechanisms by which caspase activation influence the death of lymphoma cells. In the light of our recent findings and similar observations of other investigators, it is likely that lymphoma cells possess the complete caspase machinery required for the apoptotic process. Inhibition of caspases activation appears as a potential mechanism to explain apoptotic defects of malignant B-cells, and thus may constitute the basis for new cancer therapies. PMID- 11378539 TI - Therapy related leukemias: susceptibility, prevention and treatment. AB - Acute leukemia is the most frequent therapy-related malignancy. Together with the increasing use of chemo- and radiotherapy, individual predisposing factors play a key role. Most of secondary leukemias can be divided in two well-defined groups: those secondary to the use of alkylating agents and those associated to topoisomerase inhibitors. Leukemias induced by alkylating agents usually follow a long period of latency from the primary tumour and present as myelodysplasia with unbalanced chromosomal aberrations. These frequently include deletions of chromosome 13 and loss of the entire or of part of chomosomes 5 or 7. The loss of the coding regions for tumor suppressor genes from hematopoietic progenitor cells is a particularly unfavourable event, since the remaining allele becomes susceptible to inactivating mutations leading to the leukemic transformation. The tumorigenic action of topoisomerase inhibitors is on the other hand due to the formation of multiple DNA strand breaks, resolved by chromosomal translocations. Among these, chromosome 11, band q23, where the myeloid-lymphoid leukemia (MLL) gene is located, is often involved. Frequent partners are chromosomes 9, 19 and 4 in the t(9;11), t(19;11) and t(4;11) translocations. Younger age, a mean period of latency of 2 years and monocytic subtypes are characteristic features of this type of leukemia. Among patients at risk for secondary leukemia, those with Hodgkin's disease are the most extensively studied, with the major impact of alkylating agents included in the chemotherapy schedule. The same is true for non Hodgkin's lymphoma, while in multiple myeloma and acute lymphoblastic leukemia determinants are the dose of melphalan and of epypodophyllotoxin, respectively. Patients with breast, ovarian and testicular neoplasms are also at risk, in particular if trated with the association of alkylating agents and topoisomerase II inhibitors. According to the EBMT registry, in patients with lymphoma treated with high-dose therapy and autologous stem cell transplantation the cumulative risk of inducing leukemia at 5 years is 2.6%. Among treatment options, supportive therapy is indicated in older patients, while allogeneic stem cell transplantation, related or matched-unrelated, is feasible in younger patients. These data indicate the need for the identification of predisposing factors for secondary leukemia. In particular, frequent follow-up of patients at high-risk should be performed and any peripheral blood cytopenia should be considered suspicious. Whenever possible, the exclusion of drugs known to be leukemogenic from the treatment schedules should be considered, especially in young patients. PMID- 11378540 TI - Natural killer cells and the syndrome of chronic natural killer cell lymphocytosis. AB - Natural killer (NK) cells provide anti-infectious, anti-neoplastic, and immunomodulatory function effected by both cytokine production and direct cellular cytotoxicity that is not major histocompatibility complex-restricted. NK cells lack truly specific cell surface determinants as well as antigen-specific receptors. Recent information suggests a variety of receptor-ligand interactions that underlie recognition and treatment of target cells by NK cells. Primary NK cell disorders in humans are currently classified into NK cell lymphomas and chronic NK cell lymphocytosis (CNKL). In this review, we summarize current understanding of the biology of NK cells and describe the clinical manifestations of CNKL. PMID- 11378542 TI - Preliminary experience with a new chemotherapy regimen for adults with acute lymphoblastic leukemia. AB - We treated 11 consecutive adult patients who presented with acute lymphoblastic leukemia to the University of Chicago with a novel, intensified chemotherapy regimen to evaluate the feasibility and toxicity of this program. Following a 5 drug induction therapy (course A), patients received sequential courses (B and C) of high-dose antimetabolite therapies, in part to replace cranial irradiation for CNS prophylaxis. All patients achieved a complete remission. The regimen was designed with a goal of administering all post-remission therapy in the outpatient setting. With the exception of the initial induction course (A), 3(1/3)8 total patient courses were administered in the outpatient clinic. As expected, the induction and consolidation courses (A and B) of this regimen were associated with grade 4 hematologic toxicity and significant but manageable infectious toxicity. Another novel aspect of the regimen included a combined intravenous and oral dosing schedule designed to sustain methotrexate levels > or = 1.0 microM for 30 hours (course C). There was minimal toxicity with the CNS prophylaxis/high-dose methotrexate course (C). Methotrexate levels at 30 hours ranged from 0.31-4.0 microM, with a median of 1.0 microM (n=37). This study demonstrates that this novel post-remission regimen is feasible in adults and that high concentrations of methotrexate can be sustained in the outpatient setting. The Cancer and Leukemia Group B is presently evaluating the efficacy of this regimen in a large phase II trial (CALGB study 19802). PMID- 11378541 TI - Allogeneic transplantation for patients with advanced acute leukemia: a single center retrospective study of 92 patients. AB - Allogeneic transplantation is a well recognized treatment strategy of leukemia. However, its use in advanced leukemia patients is a subject of some debate especially when donors are not HLA-identical siblings because of the toxicity and cost of the procedure. We reviewed retrospectively the outcome of patients (pts) who received allogeneic transplantation for advanced acute leukemia in our center between 09/86 and 11/97. Thirty-six pts (study group) who lacked a matched sibling donor received partially matched related donor (n=14: PMRD group) or matched unrelated donor transplantation (n=22: MUD group). Fifteen pts had AML and 21 ALL. Seventeen pts (47%) were in CR>1, 13 pts (36%) had refractory disease and six pts (17.7%) were in untreated relapse. The outcome was compared to that of 56 patients (AML: 45.5 %, ALL: 55.5 %, CR>1: 49.9 %, refractory disease: 37.5 %, untreated relapse 19.6 %) who received allogeneic transplantation from a matched sibling donor (control group). Various conditioning regimens and GVHD prophylaxis were used. The actuarial incidence of grade II to IV acute GVHD was significantly higher in the study group (57%) than in the control group (34%) (p=0.047). The actuarial risk of relapse at three years was 21% +/- 22% in the study group versus 65% +/- 16% in the control group (p= 0.04). The actuarial probability of transplant-related mortality at 3 years is 64 +/- 16% for the study group and 25 +/- 11% for the control group (p=0.001). The leading cause of death in the study group was infection (30%) followed by acute GVHD and relapse. Relapse was the major cause of death in the control group (54%), followed by infection, interstitial pneumonia, veno-occlusive disease and GVHD. The OS and probability of leukemia-free survival at 3 years were 28 % +/- 15% (95% CI) and 27% +/- 15% (95% CI) in the study group. The overall survival and probability of LFS at 3 years were respectively 28 +/- 12% (95% CI) and 23 +/- 12% (95% CI) in the control group (p = 0.08 and p=0.11 respectively). In multivariate analysis, transplant-related mortality was higher in the study group (p=0.04) and lower if both donor and recipient were seronegative for CMV (p=0.007). OS was significantly higher for seronegative couples (p=0.0001), and when CR was achieved before BMT (p=0.0022). These results suggest that all efforts in this field should be directed on lowering the transplant related mortality for non geno-identical transplants and the relapse rate in geno-identical transplants. PMID- 11378543 TI - Results of therapy with interferon alpha and cyclic combination chemotherapy in patients with philadelphia chromosome positive chronic myelogenous leukemia in early chronic phase. AB - The objective of the study was to investigate the toxicity and efficacy of cyclic combination therapy offered to patients with Ph-positive CML having a sub-optimal response to IFN-alpha. Patients in early chronic phase CML were treated with IFN alpha at 5MU/m(2) daily. Patients who did not achieve cytogenetic response after 6 months of IFN-alpha therapy, or Ph-suppression to less than 35% Ph-positive cells (partial cytogenetic response) after 12 months of therapy were offered cyclic intensive chemotherapy every 6 months, with IFN-alpha maintenance between cycles. The initial 3 cycles included daunorubicin, vincristine, cytosine arabinoside (ara-C) and prednisone (DOAP). Later cycles were given with cyclophosphamide replacing daunorubicin (COAP). Of 74 patients treated, 61 (82%) achieved complete hematologic response (CHR): 51 (69%) had a cytogenetic response, which was major (Ph < 35%) in 31 (42%), and complete in 23 (31%). Fifty five patients (74%) achieved CHR by 6 months of therapy, 38 (69%; 51% of total) with a cytogenetic response - 13 (24%) had a major cytogenetic response. Seventeen patients received at least 1 course of DOAP therapy. Median survival of the overall cohort of patients was 120 months. With a median follow-up of 145 months (103+ to 155+ months), 40 patients (54%) have died. The median duration of cytogenetic response was 35 months (range 3 to 149+ months) and the estimated 10 year cytogenetic response rate was 37%. A durable complete cytogenetic response was observed in 16 patients (20%) with a median duration of 139+ months (range 12+ to 149+ months), 11 of them (15%) are now off IFN-alpha therapy for a median of 57+ months (range 12+ to 128+ months). The projected 10-year survival was 50% for the study group versus 35% for 208 patients who received other IFN-alpha based regimens at the MD Anderson Cancer Center (p<.01). In conclusion, the addition of intensive chemotherapy may improve survival in patients with CML who have not obtained an adequate cytogenetic response on an IFN-alpha-based regimen. PMID- 11378544 TI - Fludarabine and cytarabine as a sequential infusion regimen for treatment of adults with recurrent, refractory or poor prognosis acute leukemia. AB - We did a retrospective analysis on the safety and efficacy of sequential infusion fludarabine and cytosine arabinoside (ara-C) in treating refractory, recurrent or poor prognosis acute leukemia in adult patients. Forty-five adult patients with acute myelogenous leukemia (AML) or acute lymphoblastic leukemia (ALL) received a total of 68 courses of sequential continuous infusion of fludarabine for 2 days (total dose 71.5 mg/m(2) ) followed by 3 days of ara-C (total dose 7590 mg/m(2) ). Thirty-nine patients had refractory or recurrent disease, and six had other adverse prognostic features. Thirty-six patients had AML, seven had ALL, and two had CML in blastic phase. Complete remission was seen in 20 patients (44%), and partial remission in 5 patients (11%), giving a total response rate of 56%, similar for both AML and ALL. Duration of response to prior therapy did not affect the response rate. All 3 patients with Philadelphia chromosome positive ALL obtained complete remission. Median remission duration was 4.7 months (range 0.6-36.6), and median overall survival was 5.0 months (0.7-40+). Median overall survival was 10.1 months in responders. Pulmonary toxicity was seen in 8 patients, of whom 2 died from adult respiratory distress syndrome. No cardiac toxicity was observed, but 3 patients had transient cerebellar toxicity. Profound myelosuppression was seen in all patients. We conclude that the sequential infusion of fludarabine and ara-C is an effective non-cardiotoxic regimen for adults with refractory, recurrent or poor prognosis acute leukemia, may be particularly useful for resistant Philadelphia chromosome positive ALL, and may warrant further investigation in this subset. Pulmonary rather than neurological toxicity may be a unique side effect of the regimen. PMID- 11378545 TI - High remission rate in acute myeloblastic leukemia with only two days of chemotherapy. AB - Twenty five patients with AML who had neither a history of toxic exposure or myelodysplasia were treated with a remission induction regimen consisting of two pulses of chemotherapy separated by 96 hrs. Each pulse consisted of cytarabine 2gm/m(2) (at t=0 and t=12 hrs) with mitoxantrone [30mg/m(2) ] administered immediately after the second cytarabine administration. Amifostine was administered three times a week [on Monday, Wednesday, and Friday] until the outcome of therapy was known. This regimen induced complete remissions in 15 of 17 patients less than 70 years of age and in 5 of 8 patients older than 70 years. PMID- 11378546 TI - Chronic lymphocytic leukemia associated with myelodysplastic syndrome and/or chronic myeloid leukemia: evidence for independent clonal chromosomal evolution. AB - We describe the cytogenetic findings of three cases with simultaneous or sequential development of a B-chronic lymphocytic leukemia (B-CLL) and either a myelodysplastic syndrome (MDS) in 2 cases or a chronic myeloid leukemia (CML) in one case. The coexistence of these two hematologic malignancies leads to questions about their cell of origin. Through analysis of the cytogenetic abnormalities, we studied the derivation of both malignancies. The cytogenetic analyses of these three patients were simultaneously studied from both peripheral blood and bone marrow. Furthermore unstimulated short-time (USSTC) and long-time (72-96 hours) stimulated cultures (LTSC) were systematically performed. In all cases, we have demonstrated the independent bi-clonal evolution. This is the first report ever described for patients with CLPD and MDS and/or MPD shown to arise from distinct chromosomal abnormalities. PMID- 11378547 TI - Intensified preparative regimens and allogeneic transplantation in refractory or relapsed intermediate and high grade non-Hodgkin's lymphoma. AB - Between September 1986 and June 1998, 32 patients with relapsed or refractory intermediate or high grade lymphoma received intensified preparative therapy and underwent allogeneic transplantation at a single institution. Patients were considered for allogeneic transplantation if they failed to respond to initial therapy, failed to respond to salvage therapy, relapsed after autologous transplantation, had bone marrow involvement, or failed attempts to harvest autologous stem cells. Patients had a median age of 39 years and had generally received at least two chemotherapy regimens. Five year actuarial survival (S) was 16% +/- 6%; median survival was 4 months. Survival was significantly worse in patients who had received high intensity brief duration chemotherapy prior to transplantation and was also significantly worse in patients who did not receive total body irradiation (TBI). This likely reflects the fact that the patients with the most resistant disease had required local radiotherapy and could not receive TBI. While treatment related mortality played a major role in limiting the effectiveness of allogeneic transplantation, in this heavily pre-treated population of patients with resistant disease, only 39% of patients achieved a complete response following allogeneic transplantation, and in only 40% of that group was long term disease free survival achieved. PMID- 11378548 TI - Secondary prophylactic G-CSF (filgrastim) administration in chemotherapy of stage I and II Hodgkin's lymphoma with ABVD. AB - The purpose of this study was to determine the effect of granulocyte colony stimulating factor (filgrastim, G-CSF) for maintenance of chemotherapy dose intensity in patients with stage I or II Hodgkin's lymphoma treated with six cycles of doxorubicin, bleomycin, vinblastine, and dacarbazine (ABVD). Fifty-six patients with stage I or II Hodgkin's lymphoma treated with ABVD were eligible for secondary prophylactic G-CSF administration because of neutropenia (absolute neutrophil count < 1 x 10(9) /L) causing treatment delay or febrile neutropenia. Patients received 300 microg (total dose) of G-CSF (filgrastim) subcutaneously on days 3 to 7 and 17 to 21 of each cycle in order to prevent dose reduction or delay in subsequent cycles of treatment continuing the G-CSF until completion of chemotherapy. Results showed that 30 (54%) of the patients required the use of G CSF, 26 (47%) during the first or second cycle. After G-CSF administration delay in chemotherapy did not occur in 25 patients, whereas delays in the fifth or sixth cycle occurred in four patients. Despite treatment with G-CSF, one patient had febrile neutropenia. Dose intensity greater than 90% of that planned was delivered to more the 85% of patients. IN CONCLUSION: Secondary prophylactic G CSF administration was necessary in more than half of patients with stage I or II Hodgkin's lymphoma during chemotherapy with ABVD. The use of G-CSF allowed maintenance of chemotherapy schedule and dose intensity in the majority of patients. PMID- 11378549 TI - Reduced-dose chop therapy for elderly patients with non-Hodgkin's lymphoma. AB - While CHOP therapy is effective for malignant lymphoma, the optimum schedule for elderly patients remains controversial. The present study investigated the usefulness of reduced-dose CHOP therapy for elderly patients. Previously untreated patients aged 65 years or older with intermediate to high-grade non Hodgkin's lymphoma were given up to 6 courses of reduced-dose CHOP therapy at 3 week intervals. Group A patients were given (5/6) of the standard dose and Group B received 7/12 of the standard dose. Filgrastim was administered when the white blood cell count fell below 2,000/microL. Fifty-seven patients were evaluable and the scheduled therapy was completed in 37. For patients aged from 65 to 79 years and for patients older than 80 years, the complete response rate was 79.5% and 46.2%, overall 3-year survival was 58.2% and 30.4%, and event-free 3-year survival was 49.3% and 44.4%, respectively. Major toxicities (> or = grade 3) included leukopenia in 42 patients and documented infection in 7 patients. Grade 3 cardiac plus renal failure, grade 3 peritonitis due to small bowel perforation, and grade 3 liver dysfunction occurred in 1 patient each. One patient died of toxicity (grade 4 hematological toxicity and pneumonia). In conclusion, it seems that in the elderly patients with non-Hodgkin's lymphoma, response to reduced dose ((5/6) dose) CHOP therapy is comparable to that for standard CHOP in younger adults, mainly because of improved dose-intensity. PMID- 11378550 TI - Possible immortalization of Hodgkin and Reed-Sternberg cells: telomerase expression, lengthening of telomere, and inhibition of apoptosis by NF-kappaB expression. AB - Telomerase, an enzyme associated with cellular immortality, is expressed on malignant tumor cells. Deregulation of telomerase is thought to facilitate tumorigenesis and cellular immortality by providing cancer cells with unlimited proliferation capacity. Hodgkin and Reed-Sternberg (H&RS) cells are generally considered as neoplastic cells in Hodgkin's disease (HD), however, such cells are only found in a minority of HD lesions. In addition, H&RS cells with mitotic features are rare and mummified forms are occasionally encountered. There are no available data on the relationship between telomerase activity and apoptosis in HD. We studied 14 cases with Hodgkin's disease (mixed cellularity type, nine cases; nodular sclerosis type, five cases) to clarify the relationship between telomerase activity and apoptosis using in situ hybridization of human telomerase reverse transcriptase (hTERT), reverse transcriptase-polymerase chain reaction (RT-PCR) of hTERT, using extracted RNA and immunohistochemistry of nuclear factor ?B (NF-?B), and TdT-mediated dUTP-digoxigenin nick end-labeling (TUNEL) technique for apoptosis. We also analyzed the telomere length, using sorted H&RS cells. TUNEL showed a few apoptotic H&RS cells, but the cells frequently expressed hTERT, as confirmed by ISH and RT-PCR. Lengthening of the telomere of H&RS cells was noted in ten cases. In addition, H&RS cells frequently expressed NF-?B, which is known as an inducible transcription factor and inhibitor of apoptosis. Our findings of telomerase activity in H&RS cells indicate that these cells are neoplastic and are potentially immortal. In addition, NF-?B expression on H&RS cells suggests its possibility in inhibition of apoptosis of these cells. PMID- 11378551 TI - Primary central nervous system lymphomas express Vh genes with intermediate to high somatic mutations. AB - Primary central nervous system lymphoma (PCNSL) is a rare disease, especially among non-AIDS patients. Although almost all PCNSLs belong to the diffuse large B cell lymphoma (DLBL) category, its clinical course differs from that of other types of DLBL. To elucidate the histogenesis of PCNSL, we analyzed the source of the cells from its variable region (VH) sequences using the polymerase chain reaction (PCR) method to amplify the immunoglobulin heavy chain (IgH) gene of DNA extracted from paraffin sections. Fifteen patients with AIDS-unrelated PCNSL of DLBL type, (7 males and 8 females), were evaluated. Only one case showed positive evidence of EBV infection. The prognosis was very poor with a median survival of 9 months. Analysis of the VH sequences revealed that the VH4 family was used in 4 cases and the VH3 family in 2 cases. The homology with previously published germline sequences was random, ranging from 82.7-93.2%, showing intermediate to high somatic mutations. In 3 of 6 cases, the existence of intraclonal diversity was suspected. These findings suggest that PCNSLs are histogenetically derived from antigen selected B cells in the germinal center (GC) environment. PMID- 11378552 TI - Peripheral blood monoclonal B-cells predict the event free survival in multiple myeloma. AB - Reinfusion of myeloma progenitor cells may contribute to relapse of multiple myeloma after autologous stem cell transplantation. The aim of our study was to investigate whether monoclonal B-cells are present in the apheresis product and to evaluate the clinical relevance of these cells. Leukapheresis products of 55 patients were purged with anti-B-cell-Monoclonal antibodies (MoAbs) and immunobeads. Monoclonal B-cells were found in 85% of patients within the B-cell population. In one third of all myeloma patients, the majority of B-cells was represented by monoclonal myeloma progenitor B-cells, whereas in two thirds of patients monoclonal cells only represented a small part of the entire B-cell population. As shown by sequence analysis, monoclonal precursor B-cells and malignant plasma cells had the identical genetic CDR III sequence. The purging efficacy, using a negative selection system, was a median of 3 logs (range 1,5 3,5). No statistical difference in the purging efficacy was found when 3, 4 or 5 MoAbs against B-cells antigens were used. However, a tumor specific signal could be detected in the purged harvest of all patients, when the highly sensitive ASO PCR approach was used. Furthermore, we found a direct correlation between the amount of remaining monoclonal cells after negative selection and the event free survival of myeloma patients. 10/15 patients with a median of 20 x 10(3) monoclonal cells in the purged product relapsed at a median of 1,4 years, whereas only 6/24 patients with an oligoclonal pattern including a low number of remaining monoclonal cells relapsed at a median of 2,2 years. The event free survival (EFS) was statistically different between the two groups (p = 0,014). PMID- 11378553 TI - Expression and in vitro modification of parathyroid hormone-related protein (PTHrP) and PTH/PTHrP-receptor in human myeloma cells. AB - To elucidate the role of PTHrP in myeloma, we examined the expression levels of PTHrP and its receptor in human myeloma cell lines and clinical specimens from 13 myeloma cases. In vitro modification of PTHrP expression and production induced by TGF-beta and PMA in PTHrP expressing myeloma cell lines was also investigated. PTHrP expression was detected in six out of seven myeloma cell lines with an inverse correlation with the expression of its receptor, and in 10 out of 13 clinical specimens in varying degrees. The PTHrP expression and secretion into culture medium were enhanced by supplemental TGF-beta and PMA. PMA also seemed to affect PTHrP upregulation via TGF-beta activation. The fundamental role of PTHrP in bone lesions and hypercalcemia in myeloma may be important to consider even during the initial phase of the disease and particularly in the progression of bone complications with hypercalcemia. PMID- 11378554 TI - Aberrant BCR-ABL transcript with intronic insertion in a patient with philadelphia chromosome-positive chronic myeloid leukemia: implications for disease progression. AB - The BCR-ABL fusion gene is important for the leukemogenesis of chronic myeloid leukemia (CML). A relationship between types of BCR-ABL transcripts in CML and clinical features has been proposed. We present here a patient with CML who carried an aberrant BCR-ABL transcript with an intronic sequence insert. A 26 year-old woman was diagnosed as having Philadelphia chromosome (Ph) positive CML. Reverse transcription polymerase chain reaction detected an atypically large BCR ABL mRNA transcript. Sequencing revealed a 589bp insertion consisting of a 5' portion of BCR intron b2 and a 3' portion of ABL intron 1b between BCR exon b2 and ABL exon a2. Although the typical b2a2 transcript was undetectable initially, it appeared after intensive chemotherapy. The aberrant transcript presumably arose as a result of a lack of splicing, and chemotherapy might modify the disease course by selecting the subpopulation of the CML clone expressing typical BCR-ABL mRNA dominantly. PMID- 11378555 TI - Cauda equina compression by malignant lymphoma: long-term survival following multimodality therapy. AB - Malignant lymphoma compression of the cauda equina is rare. In the literature only a few cases have been described with occasional long term survivors. We report another case of non-Hodgkin's lymphoma presenting with cauda equina compression. Significant palliation and long-term survival were achieved after therapy by decompressive, cytoreductive surgery; local irradiation; and chemotherapy. PMID- 11378556 TI - Diffuse large cell lymphoma and t(8;22) (q24;q11) in a patient with idiopathic CD4+ T-lymphopenia. AB - We describe a unique case of a patient with a three-year history of idiopathic CD4(+) T cell lymphopenia (HIV negative) who presented with stage IV diffuse large cell non Hodgkin's lymphoma with t(8;22). Despite the severe lymphopenia, the patient tolerated intensive chemotherapy well and at 18 months, remains in complete remission. PMID- 11378557 TI - Simultaneous occurrence of kaposi's sarcoma and chronic myelogenous leukemia. AB - We report here a 75-year-old man from South France who developed Kaposi's Sarcoma (KS) 5 months after diagnosis of Philadelphia-chromosome positive chronic myelogenous leukemia (CML). He was found positive for HHV-8 by PCR, negative for both HIV 1 and HIV 2 by serology, and had a normal CD4/CD8 ratio. Favourable evolution of both CML and KS has been obtained with vinblastine and interferon alpha treatment. The patient is currently alive in complete remission of SK and major cytogenetic remission of CML with a 48 month follow-up. Since no immune deficiency could be documented in the patient, this rare observation suggests that CML may have triggered the onset of SK through cytokine release. PMID- 11378558 TI - t(2;5) associated with a histiocytic-monocytic neoplasm. AB - We describe a patient with a neoplasm derived from the histiocytic-monocytic lineage associated with t(2;5) detected by FISH. The patient presented with bone marrow involvement, no organomegaly and subsequently developed a leukaemic picture. The clinical course was aggressive and the patient died four months from diagnosis. Cell morphology, immunophenotype (CD30-, EMA-, Lisozyme+, cy CD68+ and CD45+) and DNA analysis showing germ-line configuration of the Ig/TCR chain genes ruled out the diagnosis of anaplastic large cell lymphoma (ALCL). This unusual case ilustrates that t(2;5) is not exclusive for ALCL but may be found in a few cases of rare neoplasms derived from the histiocytic-monocytic cells. PMID- 11378559 TI - Pneumocystis carinii pneumonia with an atypical granulomatous response in a patient with chronic lymphocytic leukemia. AB - We have recently seen a patient who developed Pneumocystis carinii pneumonia (PCP) in the course of treatment for chronic lymphocytic leukemia (CLL). This case showed uncommon pathological findings with extensive formation of granulomatous lesions. Despite advanced CLL associated with poor B-cell function, she responded well to anti-PCP treatment. In contrast to B-cell function, the T cell functions were well preserved in vitro, and the numbers of peripheral CD4- and CD8-positive cells were normal, and T-cell functions were normal. These findings suggest that the production of granulomatous lesions to PCP may have been associated with the patients' immune status, and that it may constitute a good indicator in PCP infection in patients with underlying hematological malignancy. PMID- 11378560 TI - Primary effusion lymphoma (PEL) in HIV-negative patients--a distinct clinical entity. AB - Primary effusion lymphoma (PEL) is a recently described rare type of non Hodgkin's lymphoma occurring almost exclusively in HIV infected people. Human herpesvirus 8 (HHV-8), has been linked with PEL, and a causative relationship has been suggested. In the vast majority of PEL cases Epstein-Barr virus (EBV) has been found in the tumour cells. We describe here an elderly human immune deficiency (HIV) seronegative man with intractable chest pain and pleural effusion. The diagnosis of malignant lymphoma was suggested cytologically and confirmed histologically following pleural biopsy. No lymphadenopathy or organ involvement with lymphoma was found. Systemic chemotherapy with a modified CHOP regimen with G-CSF support gradually led to the resolution of the chest pain and ultimately resulted in a complete clinical remission (CCR). The presence of HHV-8 was demonstrated by PCR using paraffin-embedded tissue samples from the involved pleura, whereas EBV-associated genetic material was absent. The patient remained in CCR for 18 months and died of an unrelated cause (cerebrovascular event). Only 11 other cases with clinical and virological features similar to those of our patient have been reported in the literature. Analysis of these rare cases suggests HIV-negative EBV-negative PEL to be a distinct clinical entity with epidemiological features resembling classical KS and supports an EBV-independent role for HHV-8 in the pathogenesis of PEL. PMID- 11378561 TI - Primary extranodal mucosa associated lymphoid tissue (MALT) lymphoma of the prostate. AB - Low-grade B-cell mucosa associated lymphoid tissue (MALT) lymphoma can rarely present primarily in extranodal sites other than stomach, which is the most common site for it. Until now only four cases of primary prostatic MALTomas have been described in the literature and we report the fifth. We describe a case of primary prostatic MALToma in a 67-year-old man and the diagnosis was made on the trans-urethral resection specimen of the prostate. As the disease was limited to prostate (stage IEA), the patient was treated with external beam radiation therapy to a total dose of 4400cGy in 22 fractions. The patient achieved completed remission and has remained free of disease in the following 36 months. PMID- 11378562 TI - Prolonged molecular and clinical remission after treatment of a patient with follicular lymphoma with rituximab. AB - Rituximab is a chimeric anti-CD20 monoclonal antibody that has approval for single agent therapy in the treatment of relapsed/refractory low grade or follicular non-Hodgkin's Lymphoma. In published phase II trials, molecular remissions of PCR detectable t(14;18) disease in the peripheral blood have been reported in up to 62% of patients by three months. We report a case of a patient who achieved prolonged clinical and molecular remission following a single four week course of Rituximab that has exceeded any previous remission achieved with chemo-radiotherapy. The implications of molecular remission as a surrogate of clinical remission and molecular relapse as a harbinger of clinical relapse are reviewed and discussed. PMID- 11378563 TI - Secondary myeloid/natural killer cell acute leukemia following T-cell lymphoma. AB - A 56-year-old woman was treated with combination chemotherapy and radiation therapy for peripheral T-cell lymphoma. Following complete remission for a period of 6 months, she returned again with marked leukocytosis. Leukemic cells were characterized by scanty cytoplasm with fine azurophilic granules, and were highly positive for myeloperoxidase and sudan black-B. Immunophenotypic analysis revealed that blast cells were positive for myeloid antigens (CD13, CD33), and natural killer (NK) cell antigen (CD56), but negative for T-cell antigens (CD2, CD5, CD7), B-cell antigens (CD19, CD20), CD34, and HLA-DR. The case was diagnosed as secondary myeloid/NK cell acute leukemia following non-Hodgkin's lymphoma. Despite aggressive chemotherapy against leukemia, she died of multiorgan failure 7 months following onset of leukemia. We present, to the best of our knowledge, the first published report of what seems to be a secondary myeloid/NK cell acute leukemia following T-cell lymphoma. PMID- 11378564 TI - MALT lymphoma originating in breast and uvula. AB - A case of marginal zone B cell lymphoma of MALT type arising in the uvula and breast is reported. The patient, a 30-year-old woman who delivered a child and lactated in 1997, was suffering from Sjogren syndrome (SS). She was diagnosed with MALT lymphoma after a biopsy of the right breast and uvula. To investigate the relationship of the delivery, lactation and MALT lymphoma, we examined the immunohistochemical analysis of hormone receptors. As a result, lymphoid cells of the breast were stained with anti-progesterone receptor antibodies in the cytoplasm. Consequently, the MALT lymphoma of the uvula appeared to be associated with SS. Moreover, hormones such as progesterone may have influenced the breast involvement of MALT lymphoma in our case. PMID- 11378565 TI - Current status of retroviral vector mediated gene transfer into human hematopoietic stem cells. AB - Genetic modification of hematopoietic stem cells (HSCs) has been proposed as a treatment strategy for a variety of hematologic diseases, tracking marked cells or conferring resistance to chemotherapeutic agents. Despite early enthusiasm, the results of clinical studies involving gene transfer into HSCs has not resulted in therapeutic benefits for the vast majority of treated patients. This review describes the limitations and advances that have been made in the areas of gene transfer vectors, identification of the appropriate HSCs to target for genetic modifications and the methods used to perform gene transfer. PMID- 11378566 TI - Interleukin-12 gene therapy vaccines: directing the immune system against minimal residual leukemia. AB - Current overall survival rates for patients with AML remain poor and there is need for novel therapeutic approaches. One such approach is to use the patient's own immune system to eliminate minimal residual disease. Recent advances in genetic manipulation of tumor cells, together with a better understanding of the immune mechanisms controlling the host-tumor relationship have led to a flurry of preclinical and clinical studies on tumor cell vaccines. Here we present a brief overview of genetic manipulation of tumor cells, and highlight important principles of cancer immunity and cancer vaccines. Special emphasis is given on recent work on the role of interleukin-12 (IL-12) based vaccines in murine AML. These studies have shown that vaccines with AML cells, genetically modified to secrete IL-12, are potent stimulators of the immune system and lead to the development of both prophylactic and therapeutic anti-leukemia immunity. PMID- 11378567 TI - Is myelodysplastic related acute myelogenous leukemia a distinct entity from de novo acute myelogenous leukemia? Potential for targeted therapies. AB - Acute myelogenous leukemia (AML) can be separated by whether the presentation was proceeded by a myelodysplastic (MDS related AML) or developed de novo (dAML). Clinically, MDS related AML (mAML) has been considered to have a worse prognosis that dAML. The objective of this literature review was to identify unique biologic features of mAML. Compared to dAML, mAML is characterized by an altered immunophenotype (increased frequency of CD34, CD11b and CD25), lack of leukemic progenitor cell suppression due to TGFbeta1, increased bcl-2 expression, presence of inducible nitric oxide synthase, lower levels or mrp transcripts and increased expression of p53. Possible interpretations of these differences between mAML and dAML are presented. Implications for mAML directed therapy are discussed. PMID- 11378568 TI - Autoimmune lymphoproliferative syndrome type III, an indefinite disorder. AB - Autoimmune Lymphoproliferative Syndrome (ALPS) is a childhood disorder characterized by chronic nonmalignant lymphoproliferation and autoimmunity. Although the pathogenesis is not fully understood, deficient Fas mediated apoptosis appears to be an important factor. This deficiency can be caused by a mutation of the APTI gene (ALPS type Ia), of the FasL gene (ALPS type Ib), or of the Caspase-10 gene (ALPS type II). In one sub population of patients, no mutations have been identified as yet (ALPS type III). According to published data, the latter group is much smaller than the group of patients with ALPS type Ia. However, because of the variability of the clinical presentation and the absence of a known genetic defect, this disease is difficult to diagnose, the more so as few data have been reported on these patients. Thus, ALPS type III could be more common than believed until now. In this review we provide evidence for this hypothesis. PMID- 11378569 TI - Activating mutations of c-kit at codon 816 confer drug resistance in human leukemia cells. AB - An improved understanding of how leukemia cells grow and become resistant to treatment remains critical for developing more effective therapies. We have identified activating mutations of c-kit at codon 816 (Asp(816) ) from a revertant of the cytokine-dependent acute myeloid leukemia (AML) cell line, MO7e (D816H), and de novo childhood AML (D816N). Following transduction of the mutant c-kit cDNAs, MO7e cells acquire a growth advantage and resistance to apoptosis in response to chemotherapeutic drugs and ionizing radiation, in addition to cytokine-independent survival. Although stimulation of mutant c-kit-bearing MO7e cells with stem cell factor (SCF), a ligand for c-Kit, does not have a significant effect on cell proliferation, SCF further inhibits apoptosis induced by cytotoxic agents. These results suggest a potentially important role of Asp(816) mutations of c-kit in both malignant cell proliferation and resistance to therapy. PMID- 11378570 TI - Interferon-alpha combined with cytarabine in chronic myelogenous leukemia - clinical benefits. AB - During the last decade, several studies have evaluated the treatment of chronic phase chronic myeloid leukemia (CML) with a combination of interferon (IFN)-alpha and low- dose cytarabine (Ara-C). This combination therapy has been shown to be superior compared to monotherapy with IFN-alpha in randomized studies with regard to hematologic and cytogenetic remissions. However, the survival benefit is small, and the toxicity of the combination therapy is high. This paper reviews the published studies on IFN-alpha/low-dose Ara-C for the treatment of chronic phase CML and discusses the value of the combination therapy. PMID- 11378571 TI - Hodgkin's disease in patients infected with human immunodeficiency virus: frequency, presentation and clinical outcome. AB - We report the frequency, presenting characteristics, progression-free survival, event-free survival, overall survival and AIDS-free survival of patients with previously untreated Hodgkin's disease (HD) in the setting of infection by human immunodeficiency virus (HIV). To accomplish this we retrospectively reviewed all untreated patients presenting to the University of Texas M.D. Anderson Cancer Center between July 1985 and August 1999 with HD and HIV infection. All available records were reviewed to determine presentation, clinical characteristics, treatment outcome, progression-free survival and overall survival. We identified 887 patients with HD and 3,500 with Non-Hodgkin's Lymphoma (NHL). The ratio of NHL to HD in HIV-negative versus HIV-positive patients was 3.9 versus 6.9, respectively. There were 14 HIV-positive patients with HD and 97 with NHL. The median age of the HIV-positive HD patients was 33 years, and 13 were male. Three patients had Acquired Immune Deficiency syndrome (AIDS) at the time of HD diagnosis, and seven had B-symptoms. Ann Arbor stage was I in one, II in three, III in four and IV in six patients. Mixed cellularity histology was seen in eight, bone marrow involvement in five and extranodal disease in seven patients. Four patients had elevated serum lactate dehydrogenase, three low serum albumin, and nine elevated serum beta2-microglobulin, The median CD4 count was 160/microl. Eleven patients received ABVD or equivalent regimens, followed by radiotherapy in five. One patient was treated with COPP and radiotherapy, one with NOVP and radiotherapy and one only with radiotherapy. All patients received some antiretroviral therapy, but it was variable over the years. With a median follow up of 64 months for survivors, the projected 5-year progression-free survival was 64%, event-free survival 45%, overall survival 54% and AIDS-free survival 45%. Six patients died of complications arising from HIV infection, including one patient who had preexisting AIDS at HD presentation. Two patients died of HD, without developing other conditions diagnostic of AIDS. We conclude that in our referral patient population HIV infection is associated with preferential development of NHL rather than HD, which appears curable with standard treatment regimens. Since HIV-related deaths exceed those caused by HD, future investigation should focus on integration of chemotherapy and highly active antiretroviral therapy. PMID- 11378572 TI - Does intensive treatment with high dose chlorambucil and prednisone as first line and cladribine as second line influence the survival of the patients with chronic lymphocytic leukemia? AB - Cladribine (2-CdA) and fludarabine are the new purine analogs introduced in the treatment of chronic lymphocytic leukemia (CLL). Despite the high response rate, their influence on survival is still uncertain. The aim of this study was a retrospective analysis and comparison of the response rate and survival of CLL patients treated with high dose chlorambucil (HDChl) as first and 2-CdA as second line, with an historical group of patients never treated with purine analogs who received standard doses of chlorambucil (SDChl). We analyzed 347 patients with CLL treated between January 1985 and January 2000. Group A (190 patients) received HDChl (12 mg/m(2) ) with prednisone (P) 30 mg/m(2) daily for 7 days monthly as first line and in refractory or early relapsed patients 2-CdA (0.12 mg/kg/day) for 5 days with or without P (30mg/m(2) ) as second line. Group B (157 patients) received continuous SDChl (4-8 mg/m(2) /day) and P as first line and COP or CHOP as second line. The overall response rate (OR) for the first line was 48,4% in group A and 38,9% in group B (p=0.09). 148 patients in group A and 52 in group B received the second line treatment and the second OR was 19.6% and 13.5%, respectively (p=0.4). At the time of analysis, 124 patients died in group A and 139 in group B. Median survival was 65 months and 50 months, respectively. In group A, survival was longer in advanced Rai stage patients (p=0.001) but in early Rai stage was similar for both groups (p=0.4). We suggest that intensive treatment with HDChl as first line and 2-CdA as second line should be applied in more advanced rather than in less advanced stages of CLL until the final results of randomized clinical trials are available. PMID- 11378573 TI - Adjustment of incidence rates after an estimate of completeness and accuracy in registration of acute leukemias in a Swedish population. AB - Earlier studies have revealed undernotification of hematological malignancies in Swedish and other Cancer Registries. We present epidemiological data on AML, ALL and unspecified AL in adults diagnosed 1987-1992 in a well-defined population. Blast crises of CML were excluded. The Swedish Cancer Registry and Cause of Death Registry were compared and patient records reviewed for validation. When available, listings of pathology bone marrow reports and inpatient discharge diagnoses were utilized for casefinding. 260 cases of acute leukemias could be verified in a population of 663,135 adults, corresponding to a yearly incidence of 6.5/100,000. The median age of the patients was 69.2 years. 214 cases were AML, 38 ALL and eight unspecified AL. Undernotification in the Cancer Registry was found to be 15.4%, greater for AML and unspecified AL than for ALL. In addition the coding was not uniform, resulting in an incidence rate in adults of 5.3/100,000 for the Cancer Registry which is 18.5% lower than that of our study. A significant survival advantage was seen for notified patients. Combination of the Cancer Registry and Cause of Death Registry gave acceptable coverage, omitting only four patients. As the incidence of acute leukemias in our study is comparatively high, we hypothesize that underestimation of incidence and overestimation of survival are general problems for cancer registries. PMID- 11378574 TI - Acute leukemia during pregnancy: a single institutional experience with 17 cases. AB - We reviewed the medical records of 17 consecutive patients with concomitant acute leukemia and pregnancy seen at our institution over a 37-year period. Fifteen cases each were either newly diagnosed or classified as acute myeloid leukemia (AML). Seven diagnoses (41%) occurred in the first, 7 (41%) in the second, and 3 (18%) in the third trimester. In general, nine patients received chemotherapy while pregnant-eight in the second trimester and one in the third. The overall complete remission rate among the 13 patients with newly diagnosed AML was 69%, compared with 86% in those who were pregnant during chemotherapy. Long-term survival was documented in five of the nine complete responders. Three of four patients who elected to delay treatment until after delivery died within days of starting chemotherapy. Unintentional fetal loss occurred in four patients (29%), including two without exposure to chemotherapy. There were no instances of congenital malformation. The results from the current study confirm that pregnancy per se may not affect the outcome of chemotherapy in AML. In addition, it is suggested that treatment delays may compromise maternal outcome without improving pregnancy outcome. PMID- 11378575 TI - The relation between plasma thrombopoietin and erythropoietin concentrations in polycythaemia vera and essential thrombocythaemia. AB - Plasma thrombopoietin (TPO) was measured, by immunoenzymometric assay, in 39 patients with polycythaemia vera (PV), 33 patients with essential thrombocythaemia (ET) and 10 healthy volunteers. The mean TPO concentration was significantly higher in ET patients than in PV patients (p=0.04) and normals (p<0.001). The 6 untreated ET patients had a significantly lower mean TPO concentration compared to the 27 ET patients who were on myelosuppressive regimens (p=0.01). The mean plasma TPO for the 5 PV patients treated with phlebotomy only did not differ significantly from the corresponding mean for the 34 PV patients treated with myelosuppressive agents. Concomitantly, plasma EPO was measured in 25 of the PV patients and in 30 of the ET patients by an immunoradiometric assay with normal reference interval in adults 3.7-16 IU/L. In the 14 PV patients with EPO <3.7 IU/L mean plasma TPO did not differ significantly from the mean for the 11 PV patients with EPO >or=3.7 IU/L; neither of these two groups had plasma TPO concentrations significantly different from the mean for the control subjects. The 7 ET patients with subnormal plasma EPO had significantly lower mean plasma TPO compared to the ET patients with normal and high plasma EPO concentrations (p=0.03 and p=0.02, respectively). Also, the 16 ET patients with normal plasma EPO had significantly lower plasma TPO compared to the 8 patients with high plasma EPO (p=0.04). The mean plasma TPO for each of these three groups of ET patients was significantly higher than the corresponding mean for the controls (p<0.001 for each group). The results of the present study indicate that a relationship between plasma EPO and TPO concentrations may exist and that myelosuppressive treatment affects the TPO concentration in ET but not in PV patients. PMID- 11378576 TI - Paraffin immunoreactivity of CD10, CDw75, and Bcl-6 in follicle center cell lymphoma. AB - Follicle center cell lymphoma(FCCL) has the following immunophenotype(IP): sIg+, Pan B+, CD10+/-, CD5-, CD23-/+, CD43-, CD11c-, CD25-. In addition, reactivities of a malignant lymphoma with CDw75(LN-1) and bcl-6 are considered indicators of FCCL. Bcl-6 expression is common in Grade 1 FCCL (100%) and rare in other indolent B-cell lymphomas(BCL). In contrast, bcl-2 expression is common in FCCL (80%) and in other BCL subtypes. Since no previous study has correlated paraffin immunoreactivity(PIR) of CD10, CDw75, and bcl-6 in FCCL (Grades 1-3), this is this study's purpose. Twenty-nine FCCL's were identified and reviewed (6, Grade 1; 10, Grade 2; 13, Grade 3) from the Division of Hematopathology, St. Louis University. The diagnoses were based on morphology and immunohistochemistry(IH)(21 cases) +/- the flow cytometric IP(14 cases). The paraffin blocks were stained for CD10 (Novacastra, Vector Laboratories, Burlingame, CA), CDw75 and bcl-6 (DAKO Corporation, Carpinteria, CA). Results showed that, CD10 by paraffin IH(PIH) was positive in 23 [18(strong); 3(moderate); 2(weak)] and negative in 6(3, Grade 2; 3, Grade 3). All CD10-cases were CDw75+; 4, bcl-6+. The two CD10-, bcl-6-cases were Grade 2. CDw75 was positive in 28 cases [16(strong); 11(moderate); 1(weak)] and negative in 1 (Grade 3; CD10+, bcl-2+, bcl-6+). Bcl-6 was positive in 26 [16(strong); 6(moderate); 4(weak)] and negative in 3(Grade 2's). Thus, the sensitivity of CD10, CDw75, and bcl-6 by PIH for FCCL was 79%, 97%, and 90%, respectively. Of the three stains evaluated by PIH in FCCL, CDw75 was the most sensitive, closely followed by bcl 6. CD10 was least sensitive-79%. By combining these 3 stains, the sensitivity was 100%; thus, a combined approach is recommended. PMID- 11378577 TI - Splenic lymphoproliferative disorders in human T lymphotropic virus type-I endemic area of japan: clinicopathological, immunohistochemical and genetic analysis of 27 cases. AB - Primary splenic involvement in lymphoid neoplasms is rare and the clinicopathologic features of splenic lymphoma are not well described compared to nodal non-Hodgkin's lymphoma (NHL). Here we characterized splenic lymphomas in an human T lymphotropic virus type-I (HTLV-I) endemic area of Japan. To assess the pattern of splenic involvement and evaluate prognosis, we reviewed 27 specimens consisting of 26 splenectomies and one necropsy, which were classified using REAL classification. Cases were divided into primary splenic lymphoma in 11 patients and secondary in 16 patients. The incidence of primary splenic lymphoma was 0.3% (11 of approximately 4,000 malignant lymphomas). Primary splenic lymphomas included 7 diffuse large B cell lymphoma (DLBL), 2 follicular lymphomas (FL), and 1 each of splenic marginal zone lymphoma (SMZL) and anaplastic large cell lymphoma (ALCL). Secondary splenic lymphomas included 6 DLBL, 4 mantle cell lymphoma (MCL), 2 FL, 2 Hodgkin's disease (HD), 1 each of hairy cell leukemia and ALCL. Gross examination showed two patterns of splenic involvement; solid type (formation of large nodular mass, n=16) and disseminated type (multiple nodules with diffuse infiltration but no large nodular formation, n=10). The type could not be determined in one case. Most solid types were DLBL or FL, while MCL was of the disseminated type. Immunohistochemistry showed all but each 2 cases of ALCL and HD were of B lineage. Follow-up of 26 patients indicated that all but one patient with primary lymphoma were still alive (range, 1-89 months) and 8 of 15 patients with secondary lymphomas died due to the progression of malignant lymphoma; the survival rate at 2 years was 50% in these patients. No elevation of anti-HTLV-I antibody was found. In situ hybridization for Epstein-Barr virus (EBV) showed no reactivity of lymphoma cells, although a few small lymphocytes were positive for EBV. Hepatitis C virus was observed in 6 of 20 (30%) patients examined and 4 of 11 (36%) cases of primary splenic lymphoma. Our findings indicate that patients with primary splenic lymphoma have a favorable prognosis after splenectomy. PMID- 11378578 TI - Clinical value of serial measurement of serum C-reactive protein level in neutropenic patients. AB - C-reactive protein (CRP) is an acute phase reactant of inflammation. We evaluated the clinical value of serial measurement of CRP in neutropenic patients. CRP was shown to be useful to monitor the response to therapy for febrile episodes in neutropenia. However, we failed to show statistically significant differences in CRP levels between febrile episodes with or without clinically documented infection (p= 0.10) and with or without bacteremia (p = 0.55). Also, we could not predict febrile episodes within three days by the elevation of CRP value. The area under receiver-operating characteristic curve depicting the relationship between CRP levels and forthcoming febrile episodes was only 0.60. In conclusion, serial measurement of CRP was considered to be not useful to predict fever within three days, or to differentiate the types of infection. PMID- 11378579 TI - Chemosensitivity of TEL-AML1 fusion transcript positive acute lymphoblastic leukemia cells. AB - The TEL-AML1 fusion which results from a cryptic t(12;21) translocation is the most frequently occurring genetic abnormality in childhood acute lymphoblastic leukemia (ALL) and has been associated with an excellent treatment outcome. In the present study, we examined the FAS/BCL-2 expression profiles and chemosensitivity of primary leukemic cells from children with newly diagnosed t(12;21)TEL-AML1 fusion transcript-positive versus t(12;21)TEL-AML1 fusion transcript-negative standard risk ALL. TEL-AML1(+) ALL cells expressed higher levels of the pro-apoptotic protein Fas and lower levels of the anti-apoptotic protein Bcl2 than TEL-AML1(-) ALL cells, as determined by confocal laser scanning microscopy. TEL-AML1(+) ALL cells were more sensitive to the apoptosis-inducing effects of serum deprivation, dexamethasone and vincristine than TEL-AML1(-) ALL cells. This study provides novel mechanistic insights regarding the chemosensitivity of TEL-AML1(+) ALL cells and provides a cogent explanation for the excellent leukemia-free survival outcome of children with TEL-AML1(+) ALL treated on contemporary chemotherapy programs. PMID- 11378580 TI - Apoptosis inducing novel anti-leukemic agent, bis(4,7-dimethyl-1,10 phenanthroline) sulfatooxovanadium(IV) [VO(SO4)(Me2-Phen)2] depolarizes mitochondrial membranes. AB - Bis(4,7-dimethyl-1,10 phenanthroline) sulfatooxovanadium(IV) [VO(SO(4) )(Me(2) Phen)(2)] induces apoptosis in human NALM-6 leukemia cells. In the present report, we demonstrate that VO(SO(4) )(Me(2)-Phen)(2)-induced apoptosis is mediated through the generation of reactive oxygen species (ROS), depletion of glutathione and depolarization of mitochondrial membrane potential (DeltaPsim). Using multilaser flow cytometry methods, we further mapped out the death sequence that occurs in VO(SO(4))(Me(2)-Phen)(2)-treated leukemic cells. Triple labeling method to measure ROS, DeltaPsim and glutathione coupled with multilaser excitation flow cytometry showed that induction of ROS took place before the loss of mitochondrial permeability transition and depletion of glutathione. Correlated two parameter plots of glutathione content versus DeltaPsim showed that loss of DeltaPsim and depletion of glutathione closely follows each other. Translocation of phosphatidylserine to the outer leaflet of the cell membrane was the final step in the process before the cells became apoptotic. These results demonstrate that the mitochondrial permeability transition takes place during VO(SO(4))(Me(2) Phen)(2)-induced apoptosis and is mediated through induction of ROS and depletion of glutathione. PMID- 11378581 TI - Altered expression of nuclear non-histone protein (p44/46) in different stages of B-chronic lymphocytic leukemia. AB - Our previous data have shown some differences in electrophoretic characteristics of proteins from cellular fractions (nuclear, mitochondrial, microsomal and cytosolic) isolated from peripheral blood mononuclear cells of B-cell chronic lymphocytic leukemia (B-CLL), acute lymphoblastic leukemia (ALL) patients and healthy donors. The main differences were found in electrophoretic patterns of nuclear proteins from normal and leukemia cells, especially in the nuclear mass regions of 36-52, 58-85, and 120-180 kDa. Electrophoretically-specific nuclear non-histone protein in the molecular mass zone 44/46 kDa of cells obtained from the peripheral blood of a B-CLL patient was used to produce rabbit polyclonal antiserum. SDS-polyacrylamide gel electrophoresis as well as immunological techniques (Western blot and immunocytochemistry) indicate that the nuclear protein with a molecular mass of 44/46 kDa is specifically expressed in mononuclear cells from B-CLL patients. The expression of this particular nuclear protein seems to correlate with the progression of the leukemia. PMID- 11378582 TI - Regulation of the expression of CD45 isoforms in the Farage human B cell lymphoma line and its 10.6.1 subline. AB - Different B-cell neoplasias vary in the expression of CD45 isoforms. In the present study two sublines of a human B cell lymphoma- the original Farage line (Farage OL) and the Farage 10.6.1 subline were used to analyze the regulation of the expression of CD45 cell surface determinants. Cells of the Farage OL line constitutively expressed both CD45RO and CD45RA determinants on their cell surface. In contrast, the majority of the cells of the Farage 10.6.1 subline expressed CD45RA, and only few cells were CD45RO+. The low molecular spliced CD45 mRNA, characteristic for CD45RO was found in Farage OL cells, but was almost undetectable in Farage 10.6.1 cells. Following exposure to interleukin-4 (IL-4) a large proportion of the Farage 10.6.1 cells expressed CD45RO while in Farage OL cells the proportion of CD45RO+ was slightly reduced. The low molecular, spliced mRNA characteristic for CD45RO, was increased in Farage 10.6.1 cells following IL4 stimulation, but was slightly reduced in Farage OL cells. The molecular weight of CD45RA molecules produced by Farage cells varied from 185 kDa to 220 kDa while that of CD45RO molecules was 175 kDa. Preliminary attempts were made to determine a possible correlation between the expression of CD45RO and apoptosis in Farage cells. In both the Farage OL and Farage 10.6.1 cells the proportion of Bcl-2+ cells was lower among CD45RO+ cells than among CD45RO- cells. The present study indicates that IL4 has different effects on the alternative splicing of CD45 mRNA in two closely related B cell lymphoma lines. Thus, factors produced by the B lymphoma cells themselves may endow the cells with different patterns of responsiveness to a single stimulatory agent. PMID- 11378583 TI - Identification of novel polymorphisms in intron 7 of the human p53 gene in acute myeloid leukemia and healthy donors. AB - Screening for mutations by PCR-SSCP in exons 5 to 9 of the p53 gene in 38 bone marrow or peripheral blood specimens of patients with acute myeloid leukemia (AML) showed abnormal shifts in 9 cases. One reflected a mutation in exon 8, whereas in the other cases there were no exonic mutations identified by sequencing. As PCR primers were chosen annealing in the introns flanking the exon region, following sequencing of the encompassing introns identified 5 base substitutions at various sites in intron 7. Two of them have been described previously [1] and 3 novel polymorphisms could be identified. To determine whether these polymorphisms are linked to the pathogenesis of AML, we screened peripheral blood specimens of 26 healthy controls. We found identical base substitutions in 6 out of 26 controls. Our data suggest that these polymorphisms are not related to the pathogenesis of AML. PMID- 11378584 TI - Low-grade non-Hodgkin's lymphoma in a patient with systemic lupus erythematosus. AB - Coexistence of systemic lupus erythematosus (SLE) with low-grade non-Hodgkin's lymphoma (LGNHL) has been described occasionally in the literature with the potential pathogenetic role of monoclonal B CD5+/CD19+ cells. We report a case of LGNHL which developed 18 months after diagnosis of SLE. The monoclonal population of lymphocytes in the peripheral blood and bone marrow was CD5/CD19 negative but CD19/CD22 positive. The SLE responded well to treatment with prednisone and the course of the LGNHL was stable and cytotoxic treatment was not required. PMID- 11378585 TI - Epstein-Barr virus DNA quantitation assessed by a real-time polymerase chain reaction in a case of Burkitt's lymphoma. AB - A real-time PCR technique was used to quantify EBV DNA load in plasma, leukocytes, peritoneal cells, ascites and cerebrospinal fluid (CSF) at diagnosis and during the follow-up of a 21-year-old patient suffering from an abdominal form of EBV-associated Burkitt's lymphoma. The EBV DNA load correlated well with the clinical and biological remission status of the patient after chemotherapy confirming that EBV DNA quantitation in plasma and leukocytes from peripheral blood can be considered as a marker of the tumor load and can be analyzed in parallel for monitoring of EBV-related malignancies. PMID- 11378586 TI - Blastic variant of mantle cell lymphoma following interfollicular Hodgkin's lymphoma. AB - Infrequently, patients are diagnosed with Hodgkin's lymphoma and a morphologically distinct lymphoma. While specific subtypes of lymphomas (including Hodgkin's lymphoma) may present diagnostic difficulties, fine needle aspiration biopsy (FNAB) is sometimes useful in the evaluation and classification of these lymphoproliferative processes. We report a case of the blastic variant of mantle cell lymphoma following Hodgkin's lymphoma, interfollicular variant. A 66-year-old woman with a history of Hodgkin's lymphoma presented with increasing contralateral cervical adenopathy three years after receiving chemotherapy. FNAB with ancillary immunophenotypic characterization identified mantle cell lymphoma, blastic variant. Subsequent excisional biopsy confirmed this diagnosis and also aided in the exclusion of recurrent Hodgkin's lymphoma. In addition to identifying the previously unreported combination of blastic variant of mantle cell lymphoma and Hodgkin's lymphoma, this case emphasizes the utility of FNAB in evaluation of new masses in patient's with a previous diagnosis of Hodgkin's lymphoma. PMID- 11378587 TI - Mantle cell leukemia, prolymphocytoid type: a rarely described form. AB - Leukemic manifestations of mantle cell lymphoma are seen in a minority of cases, usually associated with extensive tumor. Usually the neoplastic cells in the peripheral blood resemble mantle cells with a mature chromatin pattern and irregular nuclear contours, or less frequently with a more "blastic" chromatin pattern. A prolymphocytic leukemia with t(11;14)(q13;q32) has previously been reported; however, a complete flow cytometric immunophenotypic profile was lacking. Mantle cell leukemia, prolymphocytoid type with complete flow cytometric data has not previously been described and is the purpose of this report. We report such a case in a 90 year-old female who presented with an elevated white blood cell count. The diagnosis was based on flow cytometric immunophenotyping and the cytomorphology of the peripheral blood combined with cyclin D1 immunohistochemical staining of the bone marrow. We describe our findings and her clinical course in order to heighten awareness of this previously rarely described entity. PMID- 11378588 TI - Primary granulocytic sarcoma presenting with bone pain and hypergammaglobulinemia. AB - Granulocytic sarcoma (GS), is an extramedullary tumorous aggregate of neoplastic myeloid precursor cells, most often associated with acute myeloid leukemia (AML). Primary GS occurs in patients with normal bone marrow and no history of hematological disorders. It is a rare disease, which can involve any organ and mimic other tumors. A correct initial diagnosis, which can be difficult, and early treatment with chemotherapy as for AML patients results in a higher rate of complete remission. We report a case of multifocal primary GS of the bone associated with oligoclonal hypergammaglobulinemia, successfully treated with AML like induction chemotherapy followed by postinduction therapy with autologous peripheral stem cells transplantation. The possible significance of the associated hypergammaglobulinemia is discussed. PMID- 11378589 TI - Critical care pharmacist: an essential intensivist. PMID- 11378590 TI - Comparison of methods for cardiac output measurement. PMID- 11378591 TI - Troponin I versus troponin T: is there actually a debate in children with sepsis? PMID- 11378592 TI - Interleukin-10 and surgery. PMID- 11378593 TI - Outcome in meningococcemia: a relationship between quantity and quality? PMID- 11378594 TI - Cannot draw generic conclusions from a single study. PMID- 11378595 TI - Rational guidelines on the provision of analgesia, sedation, and neuromuscular blockade in critical care. PMID- 11378596 TI - Clinical assessment and management of massive hemoptysis. PMID- 11378597 TI - Conflict resolution at the end of life. AB - OBJECTIVE: Limited empirical research has examined how decisions are made when the preferences of terminally ill patients conflict with physicians' recommendations. This study sought to investigate physicians' strategies for resolving conflicts with dying patients. DESIGN: Cross-sectional, qualitative interviews. SUBJECTS: Subjects were 158 physicians caring for at least one terminally ill patient. SETTING: University medical center. MEASUREMENTS AND MAIN RESULTS: We analyzed physicians' responses to the open-ended interview questions, "How do you handle a situation when a patient wants a treatment that you believe does not provide any benefit?" and "How do you handle a situation when a patient does not want a treatment you think would be beneficial?" For patient requests of nonbeneficial treatments, physicians reported the following as important: negotiating with and educating patients (71%), deferring to patient requests for benign or uncomplicated treatments (34%), convincing patients to forgo treatments (33%), refusing patient requests for nonbeneficial treatment (22%), using family influence (16%), not offering futile treatments (13%), and referring to other physicians for disputed care (9%). Potential harm (23%) and cost of treatment (18%) were reasons cited for withholding treatments. In response to patient refusals of beneficial treatments, physicians report the following as important: negotiating with patients (59%), convincing patients to receive treatment (41%), assessing patient competence (32%), using family influence (27%), and referring to other physicians (21%). CONCLUSIONS: Physicians vary in the communication and negotiation strategies they use when their medical judgment conflicts with dying patients' preferences. Medical ethicists could play a greater role in teaching ethically important communication skills. Physicians providing care at the end of life report strategies for respecting patients that reflect graduated degrees of accommodation tailored to the costliness and riskiness of requests; they are most accepting of patient requests for benign, technically easy, inexpensive, and medically effective treatments. PMID- 11378598 TI - A prospective, randomized study comparing percutaneous with surgical tracheostomy in critically ill patients. AB - OBJECTIVE: To determine the relative cost-effectiveness of percutaneous dilational tracheostomy (PDT) and surgical tracheostomy (ST) in critically ill patients. DESIGN: Prospective randomized study. SETTING: Medical, surgical, and coronary intensive care units at Barnes-Jewish Hospital, a tertiary care medical center. PATIENTS: Eighty critically ill mechanically ventilated patients requiring elective tracheostomy. INTERVENTIONS: Randomization to either PDT performed in the intensive care unit or ST performed in the operating room. MEASUREMENTS AND MAIN RESULTS: Treatment groups were well matched with respect to age (PDT, 65.44 +/- 2.82 [mean +/- se] years; ST, 61.4 +/- 2.89 years, p = Ns), gender (PDT, 45% males; ST, 47.5% males, p = NS), severity of illness (Acute Physiology and Chronic Health Evaluation II score: PDT, 16.87 +/- 0.84; ST, 17.88 +/- 0.92, p = NS), and principle diagnosis. PDT was performed more quickly (PDT, 20.1 +/- 2.0 mins; ST, 41.7 +/- 3.9 mins, p < .0001) and was associated with lower patient charges than ST (total patient charges: PDT, 1,569 dollars +/- 157 dollars vs. ST, 3,172 dollars +/- 114 dollars; equipment/supply charges: PDT, 688 dollars +/- 103 dollars vs. ST, 1,526 dollars +/- 87 dollars; professional charges: PDT, 880 dollars +/- 54 dollars vs. ST, 1,647 dollars +/- 50 dollars; p < .0001 for all). There were no differences in days intubated before tracheostomy (PDT, 12.7 +/- 1.1 days; ST, 15.6 +/- 1.9, p = .20), intensive care unit length of stay (PDT, 24.5 +/- 2.5 days; ST, 28.5 +/- 3.1 days, p = .33), or hospital length of stay (PDT 49.7 +/- 4.2 days; ST, 43.7 +/- 3.5 days, p = .28) when we compared these two techniques. CONCLUSIONS: PDT is a cost-effective alternative to ST. The reduction in patient charges associated with PDT in this study resulted from the procedure being performed in the intensive care unit, thus eliminating the need for operating room facilities and personnel. PDT may become the procedure of choice for electively establishing tracheostomy in the appropriately selected patient who requires long-term mechanical ventilation. PMID- 11378599 TI - Effect of a catecholamine-induced increase in cardiac output on extravascular lung water. AB - OBJECTIVE: To determine the influence of dopamine- and dobutamine-induced increases in cardiac output on the extravascular lung water in an experimental model of pulmonary edema. DESIGN: Animal experimental study. SETTING: Animal experimental laboratory of a tertiary hospital. SUBJECTS: Mixed-race pigs (n = 20) weighing 28-32 kg. INTERVENTIONS: After the animals were anesthetized and tracheotomized, they were injected with 0.1 mL/kg of oleic acid, producing a pulmonary edema by increased permeability. The animals then were randomized into two groups: Group I (n = 10) received no medication to alter cardiac output and remained on mechanical ventilation during the 240 mins of the experiment; group II (n = 10) received a continuous infusion of dopamine and dobutamine to produce a cardiac output increase of >or=30% the basal value and underwent the same mechanical ventilation regimen as group I. MEASUREMENTS AND MAIN RESULTS: Hemodynamic and respiratory variables were measured at 0 (baseline) and 30, 60, 120, 180, and 240 mins after the infusion of oleic acid. At 30 mins, the cardiac output of group II (5.40 +/- 0.94 L/min) was significantly (p < .005) higher than that of group I (3.65 +/- 1.02 L/min), and a similar significant increase was recorded at all measurement times until the end of the experiment. The mean pulmonary arterial pressure was similar in both groups except that at 240 mins it was significantly higher in group I (normal cardiac output) than in group II (high cardiac output; 34.9 +/- 7.9 mm Hg vs. 27.2 +/- 3.8 mm Hg, p = .01). The extravascular lung water was calculated by gravimetric method after the death of the animal. The extravascular lung water of group I (13.8 +/- 3.6 mL/kg) did not significantly differ from that of group II (11.5 +/- 4.0 mL/kg). CONCLUSIONS: An increase in cardiac output experimentally produced by the infusion of dopamine and dobutamine does not modify the amount of extravascular lung water. PMID- 11378600 TI - Report on the development of a procedure to prevent placement of feeding tubes into the lungs using end-tidal CO2 measurements. AB - OBJECTIVE: To determine the accuracy of a technique using capnography to prevent inadvertent placement of small-bore feeding tubes and Salem sump tubes into the lungs. SETTING: Twelve-bed medical intensive care unit (MICU) in a 557-bed teaching hospital. PATIENTS: A total of 25 ventilated adult MICU patients were studied-5 in phase 1 and 20 in phase 2. DESIGN: Phase 1 tested the ability of the end-tidal CO2 (ETCO2) monitor to detect flow (and thus accurately detect CO2) through small-bore feeding tubes. A small-bore feeding tube, with stylet in place, was placed 5 cm through the top of the tracheostomy tube ventilator adapter in five consecutive patients. The distal end of the feeding tube was attached to the ETCO2 monitor. The ETCO2 level and waveform were assessed and recorded. Because CO2 waveforms were successfully detected, a convenience sample of 20 adult MICU patients who were having feeding tubes placed (13 Salem sump tubes, 7 small-bore feeding tubes) was then studied. The technique consisted of attaching the ETCO2 monitor to the tubes and observing the ETCO2 waveform throughout placement. RESULTS: The study hypothesis was supported. Of the seven small-bore feeding tubes tested, all were successfully placed on initial insertion. Placement was confirmed by absence of an ETCO2 waveform and by radiograph. Of the 13 Salem sump tubes, 9 were placed successfully on first attempt and confirmed by absence of CO2 and by air bolus and aspiration of stomach contents. ETCO2 waveforms were detected with insertion of four of the Salem sump tubes; the tubes were immediately withdrawn, and placement was reattempted until successful. CONCLUSIONS: The technique described is a simple, cost-effective method of assuring accurate gastric tube placement in critically ill patients. PMID- 11378601 TI - Mathematical coupling does not explain the relationship between right ventricular end-diastolic volume and cardiac output. AB - OBJECTIVE: To evaluate the clinical significance of mathematical coupling on the correlation between cardiac output and right ventricular end-diastolic volume (RVEDV) through measurement of cardiac output by two independent techniques. DESIGN: Prospective, observational study. SETTING: Surgical intensive care unit in a level 1 trauma center. PATIENTS: Twenty-eight critically ill surgical patients who received mechanical ventilation and hemodynamic monitoring with a pulmonary artery catheter. INTERVENTIONS: A pulmonary artery catheter designed to measure right ventricular ejection fraction (RVEF) and cardiac output by the intermittent bolus thermodilution (TDCO) method and continuous cardiac output by the pulsed thermal energy technique was placed. A computerized data logger was used to collect data simultaneously from the RVEF/TDCO system and the continuous cardiac output system. MEASUREMENTS AND MAIN RESULTS: Two hundred forty-nine data sets from 28 patients were compared. There is statistical correlation between TDCO and continuous cardiac output measurements (r = 0.95, p < 0.0001) with an acceptable bias (-0.11 L/min) and precision (+/-0.74 L/min). The correlation was maintained over a wide range of cardiac outputs (2.3-17.8 L/min). There is a high degree of correlation between RVEDV and both TDCO (r = 0.72, p < 0.0001) and independently measured continuous cardiac output (r = 0.68, p < 0.0001). These correlation coefficients are not statistically different (p = 0.15). CONCLUSIONS: The continuous cardiac output technique accurately approximates cardiac output measured by the TDCO method. RVEDV calculated from TDCO correlates well with both TDCO and independently measured continuous cardiac output. Because random measurement errors of the two techniques differ, mathematical coupling alone does not explain the correlation between RVEDV estimates of preload and cardiac output. PMID- 11378602 TI - Assessment of two hand hygiene regimens for intensive care unit personnel. AB - OBJECTIVE: To compare skin condition and skin microbiology among intensive care unit personnel using one of two randomly assigned hand hygiene regimens: a 2% chlorhexidine gluconate (CHG)-containing traditional antiseptic wash and a waterless handrub containing 61% ethanol with emollients (ALC). DESIGN: Prospective, randomized clinical trial. SETTING: Two critical care units (medical and surgical) in a large, metropolitan academic health center in Manhattan. SUBJECTS: Fifty staff members (physicians, nurses, housekeepers, respiratory therapists) working full time in the intensive care unit. INTERVENTIONS: One of two hand hygiene regimens randomly assigned for four consecutive weeks. MEASUREMENTS AND MAIN RESULTS: The two outcomes were skin condition (measured by two tools: Hand Skin Assessment form and Visual Skin Scaling form) and skin microbiology. Samples were obtained at baseline, on day 1, and at the end of wks 2 and 4. Participants in the ALC group had significant improvements in the Hand Skin Assessment scores at wk 4 (p = 0.04) and in Visual Skin Scaling scores at wks 3 (p = 0.01) and 4 (p = 0.0005). There were no significant differences in numbers of colony-forming units between participants in the CHG or ALC group at any time period. The ALC regimen required significantly less time than the CHG regimen (mean: 12.7 secs and 21.1 secs, respectively; p = 0.000) and resulted in a 50% reduction in material costs. CONCLUSIONS: Changes in hand hygiene practices in acute care settings from the traditional antiseptic wash to use of plain, mild soap and an alcohol-based product should be considered. Further research is needed to examine the association between use of antiseptic products for hand hygiene of staff and reductions in nosocomial infection rates among patients. PMID- 11378603 TI - Influence of prophylactic use of pentoxifylline on postoperative organ function in elderly cardiac surgery patients. AB - OBJECTIVE: To study the effects of pretreatment with pentoxifylline before cardiac surgery on postoperative organ function in elderly patients (>80 yrs) undergoing cardiac surgery. DESIGN: Prospective, randomized, placebo-controlled study. SETTING: Two-day clinical investigation in an intensive care unit of a university-affiliated hospital. PATIENTS: Forty elderly patients (age >80 yrs) undergoing first-time elective aortocoronary bypass grafting. INTERVENTIONS: In 20 patients, pentoxifylline (loading bolus of 300 mg followed by a continuous infusion of 1.5 mg.kg-1.hr-1 until the second postoperative day) was given after induction of anesthesia; another 20 patients received saline solution as placebo. MEASUREMENTS AND MAIN RESULTS: Concentrations of soluble adhesion molecules (soluble E-selectin, soluble vascular cell adhesion molecule-1, and soluble intercellular adhesion molecules) were measured to assess endothelial function. Liver function was evaluated by monoethylglycinexylidide test and by measuring alpha-glutathione S-transferase plasma concentrations. Renal function was assessed by measuring serum creatinine and urine concentrations of alpha-1 microglobulin. Splanchnic perfusion was assessed by monitoring intramucosal pH by using continuous tonometry. All measurements were performed before pentoxifylline infusion (T0), at the end of surgery (T1), 5 hrs after surgery (T2), and at the morning of the first (T3) and second (T4) postoperative day. Postoperative concentrations of all measured soluble adhesion molecules were significantly higher in the nontreated controls than in the pentoxifylline-treated patients. Monoethylglycinexylidide serum concentrations were significantly lower and abnormal (<50 ng/mL) postoperatively only in the untreated control patients. alpha-Glutathione S-transferase increased in both groups with a significantly higher increase in the control group (from 3.2 +/- 1.2 to 24.1 +/- 4.2 ng/mL) than in the pentoxifylline-treated patients (from 3.8 +/- 1.9 to 11.5 +/- 2.1 ng/mL). Serum creatinine was unchanged in both groups, whereas alpha-1 microglobulin increased significantly more in the control group than in the pentoxifylline-treated group. Intramucosal pH remained almost unchanged in the pentoxifylline patients (>7.35) but decreased significantly in the control group (5 hrs after surgery, intramucosal pH 7.29 +/- 0.13). CONCLUSIONS: Pretreatment of patients aged >80 yrs undergoing cardiac surgery with pentoxifylline attenuated deterioration of endothelial, renal, and liver function as seen in an untreated control group. Splanchnic perfusion also appears to be improved in the pentoxifylline-treated group. Whether pretreatment with pentoxifylline will improve outcome in this patient population remains to be elucidated. PMID- 11378604 TI - High continuous positive airway pressure level induces ventilation/perfusion mismatch in the prone position. AB - OBJECTIVE: Gas exchange in patients with adult respiratory distress syndrome is influenced by posture. The combined effect of continuous positive airway pressure and posture has not been investigated. We studied the effect of normal spontaneous breathing, and that of continuous positive airway pressure, on ventilation/perfusion distributions in healthy volunteers while they were in supine and prone positions. SETTING: Nuclear medicine department in a university hospital. DESIGN: Experimental study. SUBJECTS: Sixteen healthy volunteers. INTERVENTIONS: In the supine or prone position, the subjects inhaled a technetium labeled aerosol (technetium-99m diethylenetriamine pentaacetic acid) through a tight-fitting mask. Single photon emission computed tomography images of the lungs were obtained. The subjects then received an intravenous injection of technetium-99m-labeled macroaggregates of albumin, and an identical single photon emission computed tomography imaging was performed. In the group that received continuous positive airway pressure, an end-expiratory pressure of 10 cm H2O was applied during both inhalation and injection. MEASUREMENTS AND MAIN RESULTS: During spontaneous breathing, ventilation/perfusion distribution assessed by regression analysis was uniform (i.e., not significantly different from zero) both in supine and prone positions, with a slope of -1.5 +/- 3.5%/cm supine and 1.5 +/- 3.5%/cm prone. During continuous positive airway pressure breathing in the supine position, ventilation/perfusion had a slope of -3.4 +/- 2.4 compared with 8.3 +/- 1.1%/cm in the prone position according to analysis of spatial resolution. CONCLUSION: There was a less favorable ventilation/perfusion ratio in the prone position when the subjects were exposed to continuous positive airway pressure of 10 cm H2O. PMID- 11378605 TI - Dehydroepiandrosterone, dehydroepiandrosterone-sulfate, and cortisol concentrations in intensive care unit patients. AB - STUDY OBJECTIVE: This purpose of this study was to determine whether severity of illness, as defined by the intensive care unit (ICU) admission APACHE II (updated Acute Physiology and Chronic Health Evaluation) score, is correlated with early morning cortisol, dehydroepiandrosterone (DHEA), and/or dehydroepiandrosterone sulfate (DHEA-S) concentrations. DESIGN: Early morning concentrations of DHEA, DHEA-S, and cortisol were determined within 24 hrs of admission and compared with admission APACHE II scores. SETTING: Medical (MICU), neurologic (NICU), and surgical (SICU) intensive care units of the University of Pittsburgh Medical Center. PATIENTS: A total of 191 men and women ranging in age from 16 to 93 yrs. All had been admitted to an ICU. MEASUREMENTS AND MAIN RESULTS: Statistically significant correlations between APACHE II scores and cortisol were observed for women in the MICU and SICU (r = .68, p = .0001; r = .35 p = .017, respectively) and for men in the NICU (r = .55, p = .003) and the SICU (r = .29, p = .036). The correlations between APACHE II scores and DHEA concentration data were statistically significant for women in the MICU (r = .37, p = .047) and SICU (r = .43, p = .002), as was the correlation between APACHE II and DHEA-S concentrations among women in the SICU (r = .38, p = .008). Although not statistically significant, a similar relationship was observed in the smaller group of NICU women (r = .40, p = .099). Each correlation was essentially unchanged when adjusted for age. CONCLUSION: These data show a positive correlation between APACHE II and cortisol concentrations in all groups except the MICU men. Also evident is the positive correlation between APACHE II scores and DHEA and DHEA-S concentrations in women, but not in men. PMID- 11378606 TI - Colonization and infection of pulmonary artery catheter in cardiac surgery patients: epidemiology and multivariate analysis of risk factors. AB - OBJECTIVE: To assess the incidence and etiology of colonization and infection of pulmonary artery catheters inserted in cardiac surgery patients. To determine the influence of some variables on the risk of developing pulmonary artery catheter colonization and infection. DESIGN: Prospective observational study of pulmonary artery catheters inserted into the internal jugular vein that were in place for >48 hrs over a 13-month period. Data collected included age, gender, nature of the cardiac surgery intervention, duration of extracorporeal circulation, date of insertion and removal, subsequent infection, and curative antimicrobial therapy. End points were pulmonary artery catheter colonization with >or=10(3) colonies on quantitative cultures and pulmonary artery catheter-related bacteremia. Risk factors for colonization were determined by multiple logistic regression. SETTING: A 17-bed cardiac surgery intensive care unit in a 480-bed teaching hospital in Paris. PATIENTS: Patients undergoing cardiac surgery procedures between May 1, 1997, and May 31, 1998. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Of 164 pulmonary artery catheters inserted in 157 patients, 19 (11.6%) and 1 (0.6%) were associated with colonization (mean duration of catheterization, 7.5 +/- 2.8 days) and bacteremia, respectively. These data represent an incidence of 17.7 and 0.93 episodes per 1000 catheterization-days, respectively. Pulmonary artery catheter colonization was caused by Gram-positive cocci in 48% (67% were coagulase-negative staphylococci), Gram-negative rods in 48%, and Candida albicans in 4%. From multivariate analysis, >4 days of catheterization was the single variable associated with a significantly increased risk of pulmonary artery catheter colonization (odds ratio, 9.81; 95% confidence interval, 1.24 77.5, p = .03). CONCLUSIONS: Our data show that the risk of pulmonary artery catheter-related colonization and bacteremia is quite low despite the use of a high-risk insertion site. In cardiac surgery patient populations, a trial evaluating the impact of a systematic pulmonary artery catheter removal after 4 days is warranted. PMID- 11378607 TI - Therapy of malignant intracranial hypertension by controlled lumbar cerebrospinal fluid drainage. AB - OBJECTIVES: To evaluate the effect of controlled lumbar cerebrospinal fluid drainage in adult patients with refractory intracranial hypertension. DESIGN: Prospective, pre- vs. postintervention study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: Twenty-three patients with severe traumatic brain injury or delayed ischemia after subarachnoid hemorrhage with intracranial hypertension refractory to aggressive treatment, including repeated applications of tromethamine, hypertonic saline solution, barbiturate coma, and decompressive craniectomy. Patients were considered for controlled lumbar cerebrospinal fluid drainage if basal cisterns on computerized tomography scan were discernible. INTERVENTIONS: After institution of a lumbar drain, cerebrospinal fluid was gradually aspirated, and then, continuous cerebrospinal fluid drainage was maintained under control of intracranial pressure (ICP) and pupillary status. MEASUREMENTS AND MAIN RESULTS: ICP and cerebral perfusion pressure before and after initiation of lumbar cerebrospinal fluid drainage and related complications were documented. The neurologic outcome of the patients was assessed according to the Glasgow Outcome Scale 6 months after injury. As a result of lumbar cerebrospinal fluid drainage, all patients demonstrated an immediate and lasting decrease of ICP and a concomitant increase of cerebral perfusion pressure. Two patients temporarily showed a unilateral fixed and dilated pupil 6 and 8 hrs after onset of lumbar cerebrospinal fluid drainage, respectively. Ten patients showed a favorable outcome, four patients survived with a severe permanent neurologic deficit, one patient remained in a persistent vegetative state, and eight patients died. CONCLUSIONS: Controlled lumbar cerebrospinal fluid drainage significantly reduces refractory intracranial hypertension. The danger of transtentorial or tonsillar herniation is minimized by considering lumbar drainage in the presence of discernible basilar cisterns only. PMID- 11378608 TI - Breathing pattern and perception at different levels of volume assist and pressure support in volunteers. AB - OBJECTIVE: Volume assist (VA) amplifies the breathing effort whereas pressure support ventilation (PSV) provides a fixed, effort-independent ventilatory support. According to the concept of VA, its level should compensate for the pathologically increased (additional) elastance (Eadd). However, it is unclear whether breathing subjects prefer an exact compensation of Eadd and whether they are able to adjust the support level by themselves. DESIGN: Prospective, interventional study. SETTING: Laboratory. SUBJECTS: Twelve healthy volunteers, nine females, three males, aged 21-33 yrs. INTERVENTIONS: Increased Eadd was generated by banding of the thorax and abdomen. Volunteers breathed via a mouthpiece with VA or PSV using a positive end-expiratory pressure of 5 cm H2O (0.5 kPa). The study was subdivided into two parts. In part I, volunteers were instructed to adjust the level of VA and PSV themselves starting from three different, randomly applied levels in each mode (2, 8, 14 cm H2O or cm H2O/L; 0.2, 0.8, 1.4 kPa[/L]). In part II, 20 levels of VA and PSV (1-20 cm H2O or cm H2O/L, 0.1-2 kPa[/L]) were randomly selected by an investigator and estimated by the volunteers using a visual analog scale. Additionally, the breathing pattern was characterized. MEASUREMENTS AND MAIN RESULTS: Eadd (7.1 +/- 1.5 cm H2O/L [0.7 +/- 0.2 kPa/L], mean +/- sd) corresponded almost exactly to the "self-adjusted" VA level of part I (7.0 +/- 3.3 cm H2O/L [0.7 +/- 0.3 kPa/L]) and to the adequate level of part II (8-9 cm H2O/L [0.8-0.9 kPa/L]). The accordant PSV levels were 5.7 +/- 2.6 cm H2O (0.6 +/- 0.3 kPa) and 6-7 cm H2O (0.6-0.7 kPa). The breathing pattern was less influenced by changes of the support level with VA compared with PSV, which may explain in part the greater comfort of VA. CONCLUSIONS: We confirmed the theoretical assumption that VA should be adapted to Eadd. Furthermore, we demonstrated that conscious subjects are able to adjust the level of VA and PSV themselves. PMID- 11378609 TI - Secretory phospholipase A2 activity correlates with postinjury multiple organ failure. AB - Postinjury multiple organ failure (MOF) may result from overwhelming systemic hyperinflammation. Secretory phospholipase A2 (sPLA2) produces many inflammatory lipid mediators, and levels have been correlated with both the severity of patient injury and postinjury mortality. The objective of this study was to characterize the plasma activity of sPLA2 type IIa in severely injured patients and to determine whether the activity of this enzyme correlates with the subsequent development of MOF. PATIENTS: Seventeen severely injured patients at known risk for MOF had blood sampled on postinjury days 0, 1, 2, 3, and 5. DESIGN: sPLA2 activity was sequentially measured and correlated with MOF scores. RESULTS: Six patients (35%) developed MOF. In comparison with non-MOF patients, MOF patients had elevated sPLA2 activity beginning 36 hrs postinjury (MOF sPLA2, 2.4 +/- 0.97, vs. non-MOF sPLA2, 0.86 +/- 0.16 active units (AU); p < .05) and continuing over the ensuing 5 days. To rule out the possibility that stored blood components required for patient resuscitation was the source of sPLA2, the sPLA2 was measured in packed red blood cells, platelet concentrates, and fresh frozen plasma over the routine storage time. None of the products tested had elevated levels of sPLA2 compared with fresh plasma from healthy adult volunteers. CONCLUSIONS: We conclude that increased sPLA2 activity is associated with the development of postinjury MOF. PMID- 11378610 TI - Safety of human albumin based on spontaneously reported serious adverse events. AB - OBJECTIVE: To evaluate the safety of human albumin administered for therapeutic purposes. DESIGN: Retrospective compilation of spontaneously reported serious adverse events. SETTING: Records of serious adverse event reports received from 1990 through 1997 by nine major suppliers of therapeutic human albumin worldwide. PATIENTS: Primarily hospitalized patients. INTERVENTIONS: Administration of human albumin. MEASUREMENTS AND MAIN RESULTS: The number of 40-g doses distributed by the nine suppliers during the study period was 95.4 x 10(6), corresponding to 3.82 x 10(6) kg albumin, and reported serious adverse events totaled 123. The incidence of all serious adverse events was 1.29 per 10(6) doses (95% confidence interval, 1.07 per 10(6) to 1.54 per 10(6) doses). No patient death was judged to be probably attributable to albumin administration. The incidence of fatal serious adverse events possibly related to albumin was 5.24 per 10(8) doses (95% confidence interval, 1.70 per 10(8) to 12.24 per 10(8) doses). CONCLUSIONS: Although underreporting must be recognized as a limitation of spontaneous adverse event reports, this study encompassing approximately 100 million albumin doses provides evidence that both nonfatal and fatal serious adverse events in albumin recipients are very rare. These results provide further support for the excellent long-term safety record of human albumin. PMID- 11378611 TI - Influence of naloxone on the increased sensitivity to propofol during hypovolemia in the rat. AB - OBJECTIVE: Hypovolemia has been shown to decrease the dose requirement for propofol. This increased effect has been explained partially by an increased end organ sensitivity to the anesthetic effect of propofol. We used the opioid blocking agent naloxone to test the hypothesis that endogenous opioids may be involved in this increased sensitivity. SUBJECTS: Thirty-two chronically instrumented rats were assigned randomly to either the hypovolemia (n = 16) or the control (n = 16) group. INTERVENTIONS: After pretreatment of each rat in the two groups with either intravenous saline (n = 8) or naloxone (3 mg/kg; n = 8), an intravenous infusion of propofol (150 mg x kg(-1) x hr(-1)) was given until 5 secs of electrical suppression of the electroencephalographic signal was observed. Return of righting reflex was used to assess depth of anesthesia, and the propofol blood concentration was determined simultaneously with high performance liquid chromatography. MEASUREMENTS AND MAIN RESULTS: The mean propofol blood concentrations at the return of righting reflex were significantly lower in the hypovolemic animals compared with the controls within both naloxone treated (2.1 +/- 0.2 microg/mL vs. 2.9 +/- 0.2 microg/mL; p < .01) and saline treated (2.2 +/- 0.1 vs. 3.0 +/- 0.2 microg/mL; p < .01) rats. The mean concentrations were not different between the saline- and naloxone-treated rats either within the hypovolemic group or within the control group. CONCLUSIONS: The results of our study indicate that it is unlikely that the increased end organ sensitivity to propofol during hypovolemia is mediated by endogenous opioids, because it was not reversed by naloxone. PMID- 11378612 TI - Nebulized sildenafil is a selective pulmonary vasodilator in lambs with acute pulmonary hypertension. AB - OBJECTIVE: To determine whether inhalation of aerosolized sildenafil with and without inhaled nitric oxide (NO) causes selective pulmonary vasodilation in a sheep model of pulmonary hypertension. DESIGN: A controlled laboratory study in instrumented, awake, spontaneously breathing lambs. SETTING: Animal research laboratory affiliated with a university hospital. SUBJECT: Twenty Suffolk lambs. INTERVENTIONS: Lambs were instrumented with a carotid artery catheter, a pulmonary artery catheter, and a tracheostomy tube and studied awake. After baseline measurements, pulmonary hypertension was induced by the continuous infusion of U46619, a thromboxane A2 analog. After breathing three concentrations of inhaled NO (2, 5, and 20 ppm), lambs were divided into two groups. Group 1 (n = 7) breathed aerosols containing 1, 10, and 30 mg of sildenafil alone, and group 2 (n = 4) simultaneously breathed NO (2 and 5 ppm) and aerosols containing 10 mg of sildenafil. Hemodynamic measurements were obtained before and at the end of each drug administration. Venous admixture was calculated, and plasma cyclic guanosine monophosphate and sildenafil concentrations were measured. MEASUREMENTS AND MAIN RESULTS: Aerosols containing 10 mg and 30 mg of sildenafil selectively decreased the pulmonary artery pressure by 21% +/- 3% and 26% +/- 3%, respectively (p < .05 vs. baseline pulmonary hypertension). When 10 mg of sildenafil was inhaled while simultaneously breathing 2 ppm and 5 ppm NO, the pulmonary artery pressure decreased by 35% +/- 3% and 43% +/- 2% (p < .05 vs. baseline pulmonary hypertension). Inhaled sildenafil did not impair systemic oxygenation, increase right-to-left intrapulmonary shunting, or impair the ability of inhaled NO to reduce right-to-left shunting. CONCLUSIONS: Nebulized sildenafil is a selective pulmonary vasodilator that can potentiate the pulmonary vasodilating effects of inhaled NO. PMID- 11378613 TI - Defibrillation energy requirements and electrical heterogeneity during total body hypothermia. AB - OBJECTIVE: Determine the effects of hypothermia on defibrillation energy requirements and cardiac electrophysiology. DESIGN: Prospective randomized acute intervention trial. SETTING: Medical center animal laboratory. SUBJECTS: Fifteen domestic farm swine. INTERVENTIONS: Swine were randomized to a hypothermia group (n = 8) or a control group (n = 7). All animals were instrumented with a transvenous defibrillation system connected to a defibrillator that delivers a biphasic-truncated waveform. Values for defibrillation energy requirements were measured at baseline (normothermia, 38-40 degrees C) and during treatment with total body hypothermia (30 degrees C) or no temperature change (sham). Hypothermia was induced by circulating ice-water through anterior and posterior surgical thermal blankets. MEASUREMENTS AND MAIN RESULTS: Defibrillation energy requirement values at 20%, 50%, and 80% were determined by using an up/down method. In the hypothermia group, defibrillation energy requirement values at baseline did not significantly change during hypothermia (defibrillation energy requirements 50% = 14 +/- 2 J vs. 15 +/- 2 J, respectively). Similarly, the defibrillation energy requirement values in the control group did not change from baseline to sham phase (defibrillation energy requirements 50% = 12 +/- 1 J vs. 13 +/- 1 J, respectively). Hypothermia profoundly affected cardiac electrophysiology, decreasing ventricular fibrillation threshold by 72%, conduction velocity by 25% (p < .01), and tissue excitability, while it prolonged ventricular repolarization and refractoriness by 7.5% to 15%, respectively (p < .05). CONCLUSIONS: Total body cooling to 30 degrees C was highly arrhythmogenic, although this unstable electrophysiological state did not alter ventricular defibrillation energy requirements. These data suggest that hypothermia may be used to slow metabolic processes without concern over the ability to successfully defibrillate and treat hypothermia-induced arrhythmias. PMID- 11378614 TI - Glucocorticoid suppresses neutrophil activation in ventilator-induced lung injury. AB - OBJECTIVE: To investigate, in a rat model, the role of the Mac-1/ICAM-1 pathway and the anti-inflammatory activity of steroid in ventilator-induced lung injury. DESIGN: Prospective, randomized controlled study. SETTING: Animal investigation using Wistar rats. INTERVENTION: Rats in three randomly assigned groups of 18, a total of 54 animals, were subject to the following: Two groups received high peak inspiratory pressure (35 cm H2O) ventilation after pretreatment with methylprednisolone (high-methylprednisolone group) or pretreatment with methylprednisolone vehicle (high-vehicle group). The third group of animals received low peak inspiratory pressure (7 cm H2O) ventilation after pretreatment with methylprednisolone vehicle (low-vehicle group). Except for animals previously killed to establish baseline values, after 40 mins of mechanical ventilation, the animals in each group were killed. Some animals provided histological samples, and the rest received total lung lavage. MEASUREMENT: We measured flow cytometry of lavage fluid, cell counts of tissue samples, and pressure-volume curves before and after mechanical ventilation. RESULTS: In the groups that received high peak inspiratory pressure ventilation, both the number of neutrophils that infiltrated the lungs and the expression of Mac-1 and ICAM-1 on neutrophils and macrophages increased significantly more than in the low vehicle group. Static lung compliance was reduced in the high peak inspiratory pressure groups. In the high peak inspiratory pressure groups, there were significantly fewer neutrophils in samples from the high-methylprednisolone group (0.412 +/- 0.1 x 10(5)) than from the high-vehicle group (1.10 +/- 0.1 x 10(5); p < .05). The high-vehicle group showed greater expression of CD11b on neutrophils, but this was significantly decreased by methylprednisolone (mean fluorescence intensity: high-vehicle, 118.4 +/- 34.3; high-methylprednisolone, 25.8 +/- 4.2; p < .05). The lung mechanics measured by pressure-volume curve analysis were deteriorated less in the high-methylprednisolone group. CONCLUSION: Our study suggests that a neutrophil-endothelium interaction via the Mac-1/ICAM-1 pathway is involved in the activation and recruitment of neutrophils in ventilator induced lung injury. Activation and recruitment of neutrophils were lessened by pretreatment with methylprednisolone, which might have contributed to the improvement of lung dysfunction after mechanical ventilation. PMID- 11378615 TI - Effects of abdominal Co2 insufflation on renal and hepatic blood flows during acute hemorrhage in anesthetized pigs. AB - OBJECTIVE: To evaluate the consequences of laparoscopy during hemorrhage, we studied the respiratory, renal, and hepatic blood flow changes induced by abdominal Co2 insufflation during severe hemorrhage in anesthetized pigs. DESIGN: Prospective animal study. SETTING: University research laboratory. SUBJECTS: Anesthetized and ventilated pigs (n = 18). INTERVENTIONS: The right carotid artery was cannulated to measure mean arterial pressure. A pulmonary artery catheter was inserted to measure mean pulmonary arterial pressure and cardiac output. After a midline abdominal incision, three flow probes were positioned around the portal vein, the hepatic artery, and the renal artery to measure portal vein blood flow, hepatic artery blood flow, and renal blood flow. To induce hemorrhage, blood was withdrawn until mean arterial pressure reached 50 mm Hg. Laparoscopy was mimicked by insufflating Co2 until intra-abdominal pressure reached approximately 15 mm Hg. Measurements were collected during hemorrhage, Co2 abdominal insufflation, and the combination of both interventions. MEASUREMENTS AND MAIN RESULTS: Severe pulmonary hypertension and hypercapnic acidosis occurred during abdominal Co2 insufflation. However, the abdominal Co2 insufflation did not aggravate the cardiac output and total hepatic blood flow changes induced by acute hemorrhage, whereas the combination of hemorrhage and abdominal Co2 insufflation markedly altered renal blood flow. CONCLUSIONS: These results suggest that renal function must be monitored carefully when performing laparoscopy in trauma patients. In contrast, hepatic perfusion seems well preserved. PMID- 11378616 TI - Soluble receptors released during acute pancreatitis interfere with the detection of tumor necrosis factor-alpha. AB - OBJECTIVE: To evaluate the interfering effect of tumor necrosis factor-alpha soluble receptor when measuring circulating concentrations of tumor necrosis factor-alpha in an experimental model of acute pancreatitis. DESIGN: Randomized, controlled trial. SETTING: Experimental laboratory. SUBJECTS: Male Wistar rats. INTERVENTIONS: Acute pancreatitis was induced by intraductal administration of 5% sodium taurocholate. Saline was administered in a control group. Serums were overloaded with known amounts of tumor necrosis factor-alpha or macrophage inflammatory protein-2. MEASUREMENTS AND MAIN RESULTS: Three hours after induction, serum concentrations of free tumor necrosis factor-alpha, total tumor necrosis factor-alpha, and soluble receptor of tumor necrosis factor-alpha were measured. No detectable concentrations of free tumor necrosis factor-alpha were found in any experimental group. By contrast, significant increases in total tumor necrosis factor-alpha and soluble receptor of tumor necrosis factor-alpha were found after induction of pancreatitis. Overloading of serum with tumor necrosis factor-alpha resulted in detection of 50% of the expected concentrations of free tumor necrosis factor-alpha from control animals and only of 5% from the pancreatitis group. Overloading the serum with macrophage inflammatory protein-2 resulted in a detection of 100% of the expected concentrations in both control and treated animals. CONCLUSION: Circulating soluble receptor of tumor necrosis factor-alpha could interfere with the detection of tumor necrosis factor-alpha in some pathologies, such as pancreatitis, that are associated with increases in soluble receptor of tumor necrosis factor-alpha. PMID- 11378617 TI - Pulmonary adrenomedullin counteracts deterioration of coronary flow and myocardial performance evoked by pulmonary endothelins in experimental acute respiratory distress syndrome. AB - OBJECTIVES: We recently showed that pulmonary endothelins may affect coronary circulation under various experimental and clinical conditions. Here, we investigated the effect of pulmonary mediators on coronary tone in experimental acute respiratory distress syndrome. We focused particularly on pulmonary endothelin-1, a major vasoconstrictor in acute respiratory distress syndrome, and on adrenomedullin, a potent vasodilator that is up-regulated by inflammatory stimuli. DESIGN: Controlled experiment that used isolated organs. SETTING: Experimental laboratory. SUBJECTS: Wistar rats. INTERVENTIONS: The saline effluent from an isolated lung was used to serially perfuse the coronary vessels of an isolated heart. We compared serial perfusion after 2-hr pretreatment of lungs with vehicle or endotoxin (50 microg/mL), and we used the following drugs to elucidate the coronary response observed: the endothelin type A receptor antagonist BQ-123 (2 microM), the endothelin type B antagonist A-192621 (500 nM), the endothelin-converting enzyme inhibitor phosphoramidon (50 microM), the calcitonin gene-related peptide type-1 receptor antagonist hCGRP(8-37) (2 microM), and the adrenomedullin receptor antagonist hAM(22-52) (200 nM) (n = 6 each). MEASUREMENTS AND MAIN RESULTS: In controls, serial perfusion decreased coronary flow to 87 +/- 3% of baseline (p < .05). BQ-123 and phosphoramidon prevented this effect, whereas blockade of endothelin type B and adrenomedullin binding receptors had no effect. After endotoxin challenge, coronary flow significantly increased to 110 +/- 2%. This response was augmented by BQ-123 (124 +/- 2%) and phosphoramidon (123 +/- 3%); A-192621 had no effect. Application of hCGRP(8-37) and hAM(22-52) significantly decreased coronary flow to 81 +/- 3% and 88 +/- 2%, respectively. Flow decrease after blockade of both adrenomedullin binding receptors (73 +/- 2%) significantly deteriorated peak left ventricular pressure, to 82 +/- 6% of baseline; rate of pressure increase, to 81 +/- 5%; and rate of pressure decline, to 77 +/- 6%. Endotoxin pretreatment elevated pulmonary venous big endothelin-1 (three-fold), endothelin-1 (two-fold), and adrenomedullin (five-fold). CONCLUSION: In experimental acute respiratory distress syndrome, pulmonary adrenomedullin--via calcitonin gene-related peptide type-1 receptor and adrenomedullin receptor--outweighs the coronary vasoconstrictor impact of pulmonary big endothelin-1 exerted via endothelin type A receptors after conversion to mature endothelin-1. The consequence is prevention of flow-related deterioration of myocardial performance. PMID- 11378618 TI - Initial effect of sodium bicarbonate on intracellular pH depends on the extracellular nonbicarbonate buffering capacity. AB - OBJECTIVE: The effect of sodium bicarbonate on intracellular pH under conditions close to those in vivo, with both bicarbonate and nonbicarbonate buffering systems, is unknown. We postulated that this effect depends on the nonbicarbonate buffering capacity because the alkali-induced back-titration of these buffers results in a concentration-dependent release of CO2 in the extracellular space, leading to a decrease in intracellular pH. DESIGN: The study was conducted in two stages. First, human hepatocytes were perfused with pH 7 bicarbonate-buffered medium (5 mM HCO3-, 20 torr Pco2) containing no nonbicarbonate buffer or small amounts (5 mM 4-[2-hydroxyethyl]-1-piperazineethanesulfonic acid [HEPES]) or large amounts (20 mM HEPES) of nonbicarbonate buffer. Second, the changes in intracellular pH of hepatocytes placed in acidotic human blood (pH 7, 5 mM HCO3-, 20 torr Pco2) at three hematocrits (40%, 20%, and 5%) were measured. SETTING: Research laboratory at a medical university. SUBJECTS: Cryopreserved human hepatocytes thawed the day before the experiments. INTERVENTIONS: Sodium bicarbonate was infused for 10 mins to increase the HCO3- concentration from 5 to 30 mM. In the second part, 20 mM sodium bicarbonate was added directly to the blood bathing the cells. MEASUREMENTS AND MAIN RESULTS: The intracellular pH was measured with the pH-sensitive fluorescent dye bis-carboxyethyl carboxy fluorescein in its esterified form, acetoxy-methyl ester, by using a single-cell imaging technique. Gas analyses were performed before and during the sodium bicarbonate load. Sodium bicarbonate caused a decrease in intracellular pH with all media except the artificial medium containing no HEPES. This decrease was small in media with low nonbicarbonate buffering capacity (5 mM HEPES and 5% hematocrit blood) and large in media with high nonbicarbonate buffering capacity (20 mM HEPES and 40% hematocrit blood). The change in intracellular pH was linked closely to the change in Pco2 caused by the sodium bicarbonate. CONCLUSIONS: The effect of sodium bicarbonate on intracellular pH depends on changes in Pco2 in the medium bathing the cells. The increase in Pco2 is correlated with the extracellular nonbicarbonate buffering capacity because of the release of H+ ions coming from the back-titration of these buffers. We conclude that sodium bicarbonate may exacerbate cell acidosis under buffering conditions close to those in vivo and that the initial changes in cell pH caused by sodium bicarbonate depend on the extracellular nonbicarbonate buffering capacity. PMID- 11378619 TI - Bronchoconstrictive and relaxant effects of lidocaine on the airway in dogs. AB - OBJECTIVE: Intravenous lidocaine commonly is used to treat ventricular arrhythmias and to attenuate reflex airway constriction and intracranial pressure elevation during airway manipulation in intensive care units. There is much controversy as to the actions of lidocaine on the airway, so the aim of this study was to compare, in detail, the actions of lidocaine with those of bupivacaine and procaine on airway caliber and the associated changes in plasma catecholamine concentrations in the dog. DESIGN: Prospective, randomized, controlled experimental in vivo and in vitro study. SETTING: A university research laboratory. SUBJECTS: Mongrel dogs. INTERVENTIONS: In the first experiment, we evaluated the effects of intravenous local anesthetics--lidocaine 0-10 mg/kg (n = 7), bupivacaine 0-2.5 mg/kg (n = 7), or procaine 0-20 mg/kg (n = 7)--on basal airway tone. In second experiment, histamine (10 microg/kg + 500 microg x kg(-1) x hr(-1), n = 6), serotonin (10 microg/kg + 500 microg x kg(-1) x hr(-1), n = 7), and methacholine (0.5 microg/kg + 300 microg x kg(-1) x hr(-1), n = 7) were infused to determine the effects of lidocaine (0-10 mg/kg) on agonist induced bronchoconstriction. In addition, the actions of lidocaine on vagal nerve stimulation were examined (n = 7). MEASUREMENTS AND MAIN RESULTS: Bronchial cross sectional area at the third bronchial bifurcation of dogs was monitored continuously through a fiberoptic bronchoscope. In the first experiment, all local anesthetics produced a dose-dependent decrease in basal bronchial cross sectional area. In the second experiment, lidocaine significantly potentiated histamine and serotonin-induced bronchoconstriction. In contrast, lidocaine antagonized methacholine- and vagal nerve stimulation-induced bronchoconstriction. CONCLUSION: We have clearly demonstrated that lidocaine may produce direct bronchoconstriction and worsen some agonist-induced bronchoconstriction, but it prevents reflex airway constriction. Therefore, we suggest that this agent be used with caution in asthmatics. PMID- 11378620 TI - Propofol and methohexital have no significant effect on mucus secretion or clearance in the anesthetized dog. AB - OBJECTIVE: Anecdotal reports suggest that propofol (Diprivan) may stimulate mucus hypersecretion in patients without pulmonary disease. The purpose of this study was to evaluate the effect of methohexital or propofol anesthesia on the physical and transport properties of airway mucus in spontaneously breathing dogs. DESIGN: Randomized, controlled, crossover laboratory study. SETTING: University laboratory. SUBJECTS: Four healthy, purpose-bred female beagle dogs. INTERVENTIONS: Dogs were anesthetized with 5 mg/kg of propofol by intravenous bolus followed by a maintenance infusion at 0.4 mg x kg(-1) x min(-1) or 4 mg/kg of methohexital followed by an infusion at 0.3 mg x kg(-1) x min(-1). Premedication with 0.05 mg/kg of acepromazine was given, and either atropine (0.2 mg) or saline was given by intravenous bolus at the time of induction. Dogs were intubated but spontaneously breathing throughout the experiment. Tracheal secretions were collected after induction and again after 40 mins. MEASUREMENTS AND MAIN RESULTS: The volume of secretions collected on the endotracheal tube during the 1.5-hr experiment and on a bronchoscopy brush over 10 mins during the experiment was measured. Tracheal epithelial potential difference was measured bronchoscopically by saturated agar bridges, and tracheal mucus transport velocity was determined by timing particle transport. The dynamic viscoelasticity of collected mucus was assessed by microrheometry, and secretion mucociliary transportability was measured in vitro. There were no differences in any physical or transport properties of airway secretions that could be attributed to the anesthetic agent. Secretion volume was significantly lower (p < .05) and epithelial potential difference was significantly higher (p = .03) with atropine premedication. Despite this, there were no differences in tracheal mucus transport velocity, viscoelasticity, or secretion mucociliary transportability with either anesthetic agent or with atropine. CONCLUSIONS: This study suggests that neither methohexital nor propofol significantly affects mucus secretion or clearance in healthy dogs. PMID- 11378621 TI - Variation in therapy and outcome for pediatric head trauma patients. AB - OBJECTIVE: This study was undertaken to examine variation in therapies and outcome for pediatric head trauma patients by patient characteristics and by pediatric intensive care unit. Specifically, the study was designed to examine severity of illness on admission to the pediatric intensive care unit, the therapies used during the pediatric intensive care unit stay, and patient outcomes. DATA SOURCES AND SETTING: Consecutive admissions from three pediatric intensive care units were recorded prospectively (n = 5,749). For this study, all patients with an admitting diagnosis of head trauma were included (n = 477). Data collection occurred during an 18-month period beginning in June 1996. All of the pediatric intensive care units were located in children's hospitals, had residency and fellowship training programs, and were headed by a pediatric intensivist. METHODS: Admission severity was measured as the worst recorded physiological derangement during the period 1 yr old (16.1% vs. 6.1%; p = .002). Comparisons by insurance status indicated that observed mortality rates were highest for self-paying patients. However, patient characteristics were not associated with use of therapies or standardized mortality rates after adjustment for patient severity. There was significant variation in the use of paralytic agents, seizure medications, induced hypothermia, and intracranial pressure monitoring on admission across the three pediatric intensive care units. In multivariate models, only the use of seizure medications was associated significantly with reduced mortality risk (odds ratio = 0.17; 95% confidence interval = 0.04-0.70; p = .014). CONCLUSIONS: Therapies and outcomes vary across pediatric intensive care units that care for children with head injuries. Increased use of seizure medications may be warranted based on data from this observational study. Large randomized controlled trials of seizure prophylaxis in children with head injury have not been conducted and are needed to confirm the findings presented here. PMID- 11378622 TI - Oxygen effects on glucose meter measurements with glucose dehydrogenase- and oxidase-based test strips for point-of-care testing. AB - OBJECTIVES: To determine the effects of different oxygen tensions (Po2) on glucose measurements with glucose dehydrogenase (GD)-based and glucose oxidase (GO)-based test strips, to quantitate changes in glucose measurements observed with different Po2 levels, and to discuss the potential risks of oxygen-derived glucose errors in critical care. DESIGN: Venous blood from healthy volunteers was tonometered to create different oxygen tensions simulating patient arterial Po2 levels. Venous blood from diabetic patients was exposed to air to alter oxygen tensions simulating changes in Po2 during sample handling. Whole-blood glucose measurements obtained from these samples with six glucose meters were compared with reference analyzer plasma glucose measurements. Glucose differences were plotted vs. different Po2 levels to identify error trends. Error tolerances were as follows: a) within +/-15 mg/dL of the reference measurement for glucose levels 100 mg/dL. SETTING AND SUBJECTS: Five healthy volunteers in the bench study and 11 diabetic patients in the clinical study. RESULTS: In the bench study, increases in Po2 levels decreased glucose measured with GO-based amperometric test strips, mainly at Po2 levels >100 torr. At nearly constant glucose concentrations, glucose meter systems showed large variations at low (39 torr) vs. high (396 torr) Po2 levels. Glucose measured with GD-based amperometric and GO-based photometric test strips generally were within error tolerances. In the clinical study, 31.6% (Precision PCx), 20.2% (Precision QID), and 23.0% (Glucometer Elite) of glucose measurements with GO-based amperometric test strips, 14.3% (SureStep) of glucose measurements with GO-based photometric test strips, and 4.6% (Accu-Chek Advantage H) and 5.9% (Accu-Chek Comfort Curve) of glucose measurements with GD-based amperometric test strips were out of the error tolerances. CONCLUSIONS: Different oxygen tensions do not significantly affect glucose measured with the GD-based amperometric test strips, and have minimal effect on GO-based photometric test strips. Increases in oxygen tension lowered glucose measured with GO-based amperometric test strips. We recommend that the effects of different oxygen tensions in blood samples on glucose measurements be minimized by using oxygen-independent test strips for point-of-care glucose testing in critically ill and other patients with high or unpredictable blood Po2 levels. PMID- 11378623 TI - Early diagnosis of retroperitoneal necrotizing fasciitis. AB - OBJECTIVE: To report survival of retroperitoneal necrotizing fasciitis in an inmunocompromised patient and to demonstrate early clinical signs that may help in the prompt diagnosis and treatment of this severe infection. DESIGN: Case report and literature review. SETTING: An adult, 18-bed intensive care unit within a university hospital. PATIENT: A 38-yr-old man who had undergone an uncomplicated closed hemorrhoidectomy was readmitted to the hospital on postoperative day 5 for erythema around the hemorrhoidectomy and a dirty brown discharge from the wound. INTERVENTIONS: Early diagnosis of retroperitoneal necrotizing fasciitis, wide and repeated debridement, broad-spectrum antibiotics, and eventual abdominal wall reconstruction. MEASUREMENTS AND MAIN RESULTS: This patient manifested periumbilical and bilateral flank erythema, reminiscent of the pattern of ecchymosis seen in cases of retroperitoneal hemorrhage. The findings demonstrate a variation of Cullen's and Grey Turner's sign, most often found in patients with hemorrhagic pancreatitis. An abdominal radiograph revealed a ground glass appearance with radiolucency outlining the bladder, consistent with retroperitoneal air. The chest radiograph showed mediastinal air extending into the neck. Sharp debridement of the retroperitoneal fat, the right anterior rectus sheath, and the right anterior thigh fascia was required to gain control of the infection. Operative cultures grew a mixed flora with Eschericha coli, beta hemolytic streptococcus, and Bacteroides fragilis predominating. The hospital course was complicated by hemodynamic instability, renal failure, pneumonia, and a pelvic abscess. The patient ultimately survived and underwent abdominal wall reconstruction with mesh. CONCLUSION: Retroperitoneal necrotizing fasciitis is an uncommon soft tissue infection that is often fatal. Early diagnosis in this case was facilitated by the unique clinical findings of a modified Cullen's and Grey Turner's sign. A review of the limited available literature suggests that survival of retroperitoneal fasciitis is possible with prompt debridement and antibiotic therapy. PMID- 11378624 TI - Effect of corticosteroids on nuclear factor-kappaB activation and hemodynamics in late septic shock. AB - OBJECTIVE: To describe the underlying pathophysiologic mechanisms of the effect of corticosteroids in a patient with late septic shock. DESIGN: Case report. SETTING: The medical intensive care unit at University Medical Center Utrecht. PATIENT: An 86-yr-old female patient with late septic shock requiring mechanical ventilation and vasopressive agents. INTERVENTIONS: Administration of hydrocortisone, 300 mg daily. MEASUREMENTS AND MAIN RESULTS: Within 3 days of corticosteroid treatment, the patient could be weaned of vasopressive agents and mechanical ventilation. Serum C-reactive protein levels normalized. Nuclear factor-kappaB activation in unstimulated and in vitro lipopolysaccharide stimulated peripheral blood mononuclear cells decreased to background level within 5 days. Repeated functional tests of the hypothalamic-pituitary-adrenal axis were normal. CONCLUSION: Our data suggest that the pathophysiologic mechanism behind the clinical effects of supraphysiologic doses of corticosteroids in late septic shock is directly related to the inhibition of nuclear factor-kappaB in peripheral blood mononuclear cells. PMID- 11378625 TI - Hand hygiene for intensive care unit personnel: rub it in. PMID- 11378626 TI - Anatomy, physiology, and pathology of the prone position and postural drainage. PMID- 11378627 TI - Therapy of pulmonary hypertension: targeting pathogenic mechanisms with selective treatment delivery. PMID- 11378628 TI - The lungs as an anti-inflammatory organ of the body. PMID- 11378629 TI - Sodium bicarbonate and intracellular acidosis: myth or reality? PMID- 11378630 TI - Right ventricular volumetric measurements: New tricks for an old dog. PMID- 11378631 TI - Significance of volume loading with crystalloids. PMID- 11378632 TI - Choice of lipid-regulating drugs. PMID- 11378633 TI - [Epidemiologic study of patients with craniomandibular disorders. Report of data and clinical findings]. AB - BACKGROUND: The purpose of this retrospective study was to identify the subjective and objective characteristics of a population referred to a clinic for diagnosis and treatment of orofacial pain and/or temporomandibular disorders. METHODS: Beginning in 1993, 825 patients, consecutively referred to the University of Naples Federico II , were examined and their records entered into a database. These data were collected by trained clinicians. Diagnostic subgroups were identified following the Research Diagnostic Criteria for temporomandibular disorders (RDC). RESULTS: Seventy-nine percent of patients were female, the mean age of the sample was 31.3+/-13 years (range: 5 to 74 years), most of patients were between 15 and 39 years of age. Based on diagnostic subgroups, patients were divided into: patients affected with myogenous pain (59%), arthrogenous pain (13%), arthrogenous and myogenous pain (16%) and fibromyalgia (4%). Sixty-three percent of the sample reported recent headaches, 53% reported parafunction, and 28% reported a previous trauma. Eighty-one percent reported spontaneous pain, which was chronic in 83% of them. The majority of patients (78%) presented a relatively high cultural BACKGROUND. CONCLUSIONS: These data appear to agree with other epidemiologic studies and depict the TMD treatment-seeking population as a predominantly female population during child-bearing years with multiple chronic pain complaints. PMID- 11378634 TI - [Effects of organic acids on corrosion of orthodontic appliances]. AB - BACKGROUND: During the last years orthodontic practitioners and patients have developed an increasing interest on the possible undesired effects originated by the growing use of treatments in this field. Among the several undesirable effects, we have analyzed those due to allergic phenomena or cases of direct toxicity resulting from the emission of metallic ions contained in the metallic parts of the orthodontic equipment. One of the essential requirements of metals for orthodontic use is biocompatibility; international regulations (CE, ISO) value the biocompatibility of materials for orthodontics. The analysis of biocompatibility in orthodontic equipment cannot leave out of consideration the evaluation of ion emission. METHODS: In our research we have considered the release of metallic ions from orthodontic equipment in water solutions of organic acids kept at a constant temperature of 37 degrees C for 28 days. The evaluation has been possible by ionic absorption spectrophotometry. RESULTS: The results show that the release of metallic ions, low as an absolute value, has a particular trend in time. The nickel, chromium and copper concentrations have been higher during the first day than in the following 27 days. CONCLUSIONS: These data reassure about the use of orthodontic equipment, since the emission of ions is low, even if allergic problems not linked to the quantity of ions emitted cannot be excluded. PMID- 11378635 TI - [HLA (Human Leukocyte Antigens) and oral immunological diseases]. AB - The immunologic, pathogenetic and clinical relationships between HLA antigens and oral immunologic diseases are described. The HLA typing is useful for an early diagnosis because it allows to single out the subjects at risk in a particular studied family. PMID- 11378636 TI - [The vertical root fracture]. AB - Remembering the epidemiology and the possible etiologies of vertical root fractures, the authors distinguish between fractures that occur in vital teeth and vertical fractures of endodontically treated roots. They discuss predisposing factors, onset modality and follow-up. They describe the symptomatology and the few radiographic pathognomonic signs. They show the importance, prognostic and therapeutic purposes, of an exact differential diagnosis as regards periodontal problems. On the basis of personal experience and the literature they believe that the one certain resolutive therapy is extraction in the case of monorooted teeth and the root amputation of the fractured root in case of plurirooted teeth with a good periodontal supporting bone. They conclude by stressing the importance of a preventive global approach to reduce, among other things, the incidence of these unusual conditions that are difficult to diagnose, present no clear signs, and have an unfavourable prognosis. PMID- 11378637 TI - Endodontic, surgical and periodontal treatment of dens invaginatus. Case report. AB - The aim of this paper is to propose a single stage global treatment of endodontic, periapical and periodontal lesions in a lateral maxillary incisor with dens invaginatus. A 24 year-old woman presenting a lateral maxillary incisor with dens invaginatus in association with periapica1 and periodontal lesions underwent simultaneous surgical, endodontic and periodontal regenerative procedures. At 2, 6, 12, 18 months follow-up the radiographic healing appeared to be improved and the periapical lesion healed completely 1 year after surgical intervention. Surgery in association with endodontic and periodontal procedures represents the treatment of choice to maximize long term prognosis in cases of dens invaginatus with chronic periapical and periodontal lesions. PMID- 11378638 TI - Associated orthodontic, surgical and hematological management of Cooley s anemia. Report of a case. AB - Cooley s anaemia or b-thalassaemia or mediterranean anaemia is a blood disease characterised by malformation of the skull and long bones, which confers a typical appearance on the patient. The complete development of facial abnormalities can be prevented by an intensive blood transfusion programme or by bone-marrow-transplantation. At the present time these therapeutic strategies would be able to help these patients grow and develop, live a prolonged life and avoid bad surgical RESULTS. The aim of this study was to evaluate the feasibility of orthodontic and maxillofacial surgical corrective treatment associated with an appropriate transfusion therapy in a b-thalassemic patient. The patient enrolled in the study was affected by major b-thalassemia and diagnosis was performed as an infant. She was referred to our centre at puberty for dento-maxillofacial disorders. Clinical, haematological and radiographic evaluation permitted a complete diagnosis. She received a combined haematological, orthodontic and maxillofacial surgical treatment. Controls for evaluating the statement of results obtained were performed at different times after the end of the therapy and have shown that the therapeutic objectives had been achieved and maintained. At the present time, complete diagnostic and therapeutic haematological strategies cannot be carried out on a large scale, especially in countries where health resources are limited. This objective reason, associated with possible low patient compliance, explains why we still encounter severe facial deformities resulting from erythroid hyperplasia. Our results suggest that this facial disfigurement requires surgical and orthodontic correction by consolidated surgical-orthodontic techniques performed according to the appropriate anaemia therapy. Although this is a preliminary study, initial encouraging results show no relapse three years after the end of the therapy. PMID- 11378639 TI - [Prosthetic rehabilitation in a patient with systemic scleroderma]. AB - A 49-year-old patient with systemic scleroderma was referred to the Department of Dental prostheses for the Degree in Dentistry and Dental Prostheses at Florence University because he was unable to wear his existing complete dentures. The upper denture was not correct owing to the position of the artificial teeth and the short base, and the lower denture caused pain when used. A complete new dental prosthesis was therefore required to overcome the difficulties caused by this systemic disease. Scleroderma is a progressive disease that causes the anelasticity of the mesenchymal tissues owing to post-inflammatory fibrotic and degenerative alterations of unknown etiology. Important changes also occur in the mouth: difficulty opening the mouth, hypo- or non-extendibility of the soft perioral tissues. The construction of the complete upper and lower dentures posed a series of major technical problems compared to a patient not suffering from this pathology. The first core, the rimming of the individual core holder, calculating the vertical dimension and the assembly of the front teeth were all phases that required technical modifications and special care, so that the techniques of our School could be adapted to this particular case. The cosmetic and functional result was undoubtedly good. The patient has been wearing these dental prostheses for five years with satisfaction. PMID- 11378640 TI - [SEM study of morphologyc and incidence of accessory canals in the furcation region of permanent molars]. AB - BACKGROUND: The lateral canals in the molar furcation region could represent a connection, well separated from the apical region, between dental pulp and periodontal tissues and may be used by bacteria or toxins both in an endodontic periodontal direction and vice versa. A SEM study was carried out on the furcation region of permanent molars to evaluate the morphology and incidence of lateral canals. METHODS: A total of 53 permanent molars were selected. The microinfiltration test was performed on each sample using methylene blue 1% before being sectioned and examined using a Zeiss Opmi 9-FC stereomicroscope and SEM ISI DS 130. RESULTS: Openings of lateral canals were found in the furcation region of 50.94% of the samples examined. The mean diameter of the orifices was 130 mm. The apertures were elliptical and flared. The holes occasionally appeared to be the end of probable single or double canals, but were not always patent. CONCLUSIONS: In the light of these findings, the authors advise disinfecting and filling the dental pulp cavity during root treatment and, in the event of severe periodontal disease, evaluating the vitality of dental elements before surgery, thus ensuring a correct therapeutic approach by not overlooking the possibility of combined lesions. PMID- 11378641 TI - HCV-RNA occurrence in saliva of odontoiatric HCV seropositive patients. AB - BACKGROUND: HCV-RNA occurrence in saliva of patients suffering from C hepatitis induced to consider saliva as a possible diffusion mean of this disease. METHODS: Saliva and blood samples from 32 C hepatitis seropositive patients, followed for odontostomatologic problems in Odontoiatric Clinic of Brescia University were obtained. In every blood and saliva sample HCV-RNA concentration was evaluated following HCV-RNA 2.0 Assay (bDNA) Quantiplex test (Chiron), in Microbiology Institute of Brescia University. RESULTS: All patients showing HCV-RNA in serum presented virus in saliva also; two patients with negative HCV-RNA serum presented virus in saliva. In latter cases, we supposed that viral concentration in serum was under sensibility threshold of employed method. CONCLUSIONS: Saliva appears an easily and not invasively obtainable medium for epidemiological studies on HCV diffusion in humans. Its role in C hepatitis transmission, on the contrary, has not been cleared till now. PMID- 11378642 TI - [Evaluation of the three-dimensional stability using dental casts after orthognathic surgery]. AB - BACKGROUND: The increasingly frequent use of orthognathic surgery raises several questions on the results of operations, the resulting morphological and functional modifications, and the long-term three dimensional stability of the arches after surgery. In order to answer these questions a study was carried out using pre- and postoperative casts of the arches in a group of patients to evaluate the three dimensional stability of the correction. METHODS: Thirty-nine patients with different forms of malocclusion were selected who had undergone orthognathodontic surgery at least one year earlier. Twenty-seven patients (69.2%) received postoperative orthodontic treatment, including 9 who had undergone lower jaw surgery (33.3%), 3 maxillary surgery (11.1%) and 15 undergoing combined surgery (55.6%). Twelve patients (30.8%) did not receive postoperative orthodontic surgery, including 2 who had undergone lower jaw surgery (16.6%), 5 maxillary surgery (41.7%) and 5 undergoing combined surgery (41.7%). The two-dimensional reproductions of dental casts were studied using electronic image processing. Five different techniques were used (Bolton, Lundstrom, Howes, Staley and Reinhardt) to evaluate the dental arches. The study also evaluated whether any difference in results could be ascribed to use or absence of pre or postoperative orthodontic treatment. RESULTS: The results indicate that protocols and surgical techniques used produce dental arches that are three-dimensionally stable and the use of orthodontic treatment gives greater stability one year after surgery compared to cases treated with surgery alone. CONCLUSIONS: Major maxillary surgery should always be considered when assessing the possible solutions to malocclusions in order to correct anomalous bone formation with a reduced risk of recidivation of the correction. PMID- 11378643 TI - [Lymph node metastasis of adenoid cystic carcinoma]. AB - BACKGROUND: The adenocystic carcinoma (ACC) of the head and of the neck is rather uncommon. It is characterized by a very slow biological behaviour, which leads to locoregional recurrences and distant metastasis. Most reports concentrate on the unpredictable long-term behaviour of this tumor, whereas lymph nodes metastasis are not usually discussed in detail. The aim of this study was to evaluate the difficulties connected with the lymph node metastasis in ACC. METHODS: A group of 33 patients from 1980 to 1998, was observed. These patients have been subjected to surgical treatment in the Maxillo-Facial Surgery Department of the Policlinico-Umberto I of Rome. RESULTS: Among 33 patients with ACC, 10 patients (30%) revealed, initially or during observation, cervical lymph node metastasis, which is a most common event in male subjects (70%). The lymph node involvement is much more common in carcinomas localized in the parotid and in the sublingual glands with a solid rather than a tubular histologic pattern. Direct correlation between age and metastasis lymph node involvement have not been documented. CONCLUSIONS: Surgery for node disease has a very little impact on the locoregional and on the distant metastasis evolution of the tumor. Thus, our protocol of treatment of lymph nodes metastatic lesions follows the trend of most of the international authors, considering that the neck dissection should be performed only with a lymph adenopathy clinically evident or strongly suspected. PMID- 11378644 TI - [Biocompatibility and physico-mechanical properties of the new Venezia root canal sealer. In vivo and in vitro test according international standards]. AB - At the moment, in most countries, there are laws in force which impose to the manufacturers well regulated testing in order to investigate and guarantee an acceptable biocompatibility of medical devices before their commercialization. Many international laboratories are committed to the definition of investigation methodologies and to the evaluation of biocompatibility in order to obtain research standards, capable to provide reproducible and comparable objective quantitative data. In every country, technical committees were put together for a standardization of methodological procedures, followed by European and international technical boards which proposed and codified methodologies and investigation approaches. UNI-EN-ISO laws contain all the results and constitute a reference point for any consideration on or evaluation of the biocompatibility of a medical device. Based on these laws, we evaluated the biocompatibility and determined the physical-mechanical characteristics of the new Venezia (Cabon S.p.A.) endodontic ZOE sealer. The Subcutaneous Implant Technique according Safavi et al. (in vivo test, ISO 10993: 1-6 Biological evaluation of medical and dental materials and devices) and Autian test of Emolysis on Rabbit Erythrocytes (in vitro test) allowed us to evaluate a good biocompatibility of the new product. Furthermore, its Setting and Working time, its radiopacity, Solubility and its Flow value completely satisfy the requirements of international standards (ISO/DIS 6876 Dental root Canal Sealing Materials). We can finally deduce that Venezia fulfil the ideal functional properties of an endodontic cements. PMID- 11378645 TI - [Removal of fractured cylindrical implants]. AB - The implant fracture is a rare but significative complication which requires a specific treatment for the surgical removal of the fractured fixture and the modification of the initial prosthetic plan. On the basis of the most recent studies and of a case personally observed in which both the osteointegrated supporting implants of a maxillary overdenture were fracturated, epidemiological, etiopathogenic, diagnostic and therapeutical aspects of this complication are analyzed. Clinical situations most at risk are emphasized, including implant supported maxillary overdenture, and the major etiologic factors, represented by defects in implant design or material and physiologic or biomechanical overload. The diagnostic criteria are described along with the three therapeutical possible options: 1) removal of the fractured implant, replacement of the implant and refabrication of the prosthesis; 2) modification of the existing prosthesis leaving the fractured portion of the implant in place; 3) modification of the fractured implant and refabrication of a portion of the prosthesis. Three surgical removal technics are then illustrated (block-section, vestibular approach, occlusal approach), with the emphasis on the one choosed in the case examined, in which the ostectomy around the remaining fixture has been carried out with a trephine drill in order to contain the bone loss. PMID- 11378647 TI - Issues in the diagnosis and treatment of sarcomas. PMID- 11378646 TI - [Odontogenic chronic maxillary sinusitis: regenerative possibilities of oro antral defect]. AB - Odontogenic maxillary sinusitis often develops into a chronic picture of unilateral inflammation, progressively interesting the whole antrum mucosa. The only decisive therapy of this pathology is a surgical one that, by completely removing pathologic tissues, starts sinus restoration by a healthy respiratory mucous membrane. This tissue grows from the nasal cavity after the radical surgery (according to Caldwell-Luc), presupposing an antrum drainage through that same cavity. Nevertheless, if the inflamed area is just limited in the central and posterior recesses, and the hiatus semilunaris of the medial wall remains open, it is possible to avoid a surgical access to the nasal cavity. In this way, an alternative surgery is proposed for chronic sinusitis which, using principles of Guided Bone Regeneration, permits to completely remove pathologic tissues and to start bone reformation in the damaged alveolar. PMID- 11378648 TI - Radiographic imaging of musculoskeletal neoplasia. AB - BACKGROUND: Imaging is an integral part of the diagnosis, staging and evaluation of outcomes for bone and soft-tissue neoplasms. Each of the available imaging tools has a different role. METHODS: The authors reviewed the efficacy of the current imaging modalities in the diagnosis, staging, and follow-up of patients with musculoskeletal neoplasia. RESULTS: Plain-film radiography remains the gold standard in the differential diagnosis of bone lesions. Bone scintigraphy is an excellent screening modality, and computed tomography is especially useful in evaluating lesions of the axial skeleton. The superior soft-tissue resolution and multiplanar capabilities achieved with magnetic resonance imaging, however, has replaced the need for CT scans in many cases. CONCLUSIONS: The technological advances seen in recent years in all areas of imaging have improved the capabilities of these modalities to assist in the diagnosis, definition of tumor extent, and accurate staging of musculoskeletal tumors. PMID- 11378649 TI - Fine-needle aspiration biopsy of sarcomas and related tumors. AB - BACKGROUND: Largely due to a lack of experience, familiarity, and/or confidence, few centers rely on simple fine-needle aspiration biopsy (FNAB) for the diagnosis of sarcomas and related tumors. METHODS: The authors have reviewed their own experience in more than 200 cases of FNAB of bone and soft-tissue tumors, as well as cases reported in the literature. RESULTS: FNAB has proven to be accurate and useful in 8 consecutive years of clinical experience. No serious complications have occurred. CONCLUSIONS: FNAB is recommended as an integral part of the initial evaluation of amenable orthopaedic tumors, including sarcomas, especially in cases with classic clinical and radiographic findings. PMID- 11378650 TI - Genetic and molecular abnormalities in tumors of the bone and soft tissues. AB - BACKGROUND: Malignant transformation requires the accumulation of multiple genetic alterations such as chromosomal abnormalities, oncogene activation, loss of tumor suppressor genes, or abnormalities in genes that control DNA repair and genomic instability. Sarcomas are a heterogeneous group of malignant mesenchymal tumors of difficult histologic classification and strong genetic predisposition. This article provides a comprehensive review of the cytogenetic abnormalities observed in bone and soft-tissue tumors, emphasizing known downstream molecular changes that may play a role in oncogenesis. METHODS: The database of the National Library of Medicine was searched for literature relating to genetic and molecular mechanisms in sarcomas in general and in each of the main tumor entities. RESULTS: Recent techniques in chromosome analysis and molecular cytogenetics have improved our ability to characterize genetic changes in mesenchymal tumors. Some changes are so characteristic as to be virtually pathognomonic of particular histologic types, while others are complex, difficult to characterize, and of unknown relevance to pathogenesis. The implications to the cell of some of these abnormalities are now being recognized. CONCLUSIONS: The study of sarcomas will benefit from the information derived from genetic studies and translational research. The human genome project and new methodologies, such as computer-based DNA microarray, may help in the histogenetic classification of sarcomas and in the identification of molecular targets for therapy. PMID- 11378651 TI - Gastrointestinal stromal tumors. AB - BACKGROUND: Gastrointestinal stromal tumors (GISTs) are the most common mesenchymal tumors of the gastrointestinal tract. A relationship to the interstitial cells of Cajal (ICCs) has been proposed, and expression of CD117, the c-kit receptor present in ICCs, has been suggested as a marker for GISTs. METHODS: The English literature has been reviewed with an emphasis on histogenetic features, especially the potential relationship of GISTs to ICCs. RESULTS: GISTs are most common in the stomach (70%), followed by small intestine (20%), colon and rectum (5%), and esophagus (<5%). GISTs commonly have activating mutations in exon 11 (or rarely exon 9 and exon 13) of the KIT gene that encodes a tyrosine kinase receptor for the stem cell factor or mast cell growth factor. CONCLUSIONS: Malignant potential is best estimated by the simultaneous evaluation of several clinical parameters. The only absolute criterion for malignancy is tumor spread beyond the organ of origin at the time of diagnosis. The remarkable clinical response of tumors that express c-kit to treatment with the tyrosine kinase inhibitor STI571 is a triumph of molecular pharmacology. PMID- 11378652 TI - Unplanned surgical excision of tumors of the foot and ankle. AB - BACKGROUND: Soft-tissue tumors of the foot and ankle are relatively common and mostly benign. Thus, many malignant tumors in this region are improperly treated initially. Unplanned excisions can lead to complications that may adversely affect patient outcomes and prognosis. METHODS: A retrospective review of patients treated at our institute over a 20-year period for malignant soft-tissue tumors of the foot and ankle was performed. The effect of unplanned surgical excisions on outcomes was examined. RESULTS: When limb salvage was attempted, patients who underwent unplanned surgical excisions had more complications and more extensive surgical procedures involving free flaps, and they were more likely to require adjuvant radiotherapy. No difference in recurrence and disease free survival was evident between the two patient populations. CONCLUSIONS: Despite the lack of statistical power to demonstrate differences in recurrence and survival, unplanned surgical excisions of soft-tissue sarcomas of the foot and ankle probably adversely affect quality of patient care. Suspicious lesions should be referred to surgeons trained in oncologic principles for evaluation and treatment. PMID- 11378653 TI - Hyperthermic isolated limb perfusion for extremity sarcomas. AB - BACKGROUND: The treatment options available for extremity sarcomas are amputation or limb-sparing surgery with radiation, which may incur significant morbidity and body disfigurement. Hyperthermic isolated limb perfusion (HILP) may be an attractive option in extremity sarcomas for unresectable lesions to preserve limb function and maintain quality of life. METHODS: We report the outcomes of 5 patients who underwent HILP for unresectable primary or recurrent extremity sarcomas from 1994 to 2000 at our institution. RESULTS: All patients had initial complete clinical responses to HILP, and the limb was salvaged in 4 of the 5 patients. Complications included chronic lymphedema, neuropathic pain, and prolonged wound healing. CONCLUSIONS: HILP with melphalan is a safe and effective treatment option for selected patients with locally advanced and unresectable extremity sarcomas. The response rates are high, with limb salvage occurring in most patients. Further studies of larger groups of patients are warranted. PMID- 11378654 TI - Well-defined hypodensity in the liver. PMID- 11378655 TI - POEMS syndrome: a case report and discussion. PMID- 11378656 TI - Microencapsulation and tissue engineering as an alternative treatment of diabetes. AB - In the 70's, pancreatic islet transplantation arose as an attractive alternative to restore normoglycemia; however, the scarcity of donors and difficulties with allotransplants, even under immunosuppressive treatment, greatly hampered the use of this alternative. Several materials and devices have been developed to circumvent the problem of islet rejection by the recipient, but, so far, none has proved to be totally effective. A major barrier to transpose is the highly organized islet architecture and its physical and chemical setting in the pancreatic parenchyma. In order to tackle this problem, we assembled a multidisciplinary team that has been working towards setting up the Human Pancreatic Islets Unit at the Chemistry Institute of the University of Sao Paulo, to collect and process pancreas from human donors, upon consent, in order to produce purified, viable and functional islets to be used in transplants. Collaboration with the private enterprise has allowed access to the latest developed biomaterials for islet encapsulation and immunoisolation. Reasoning that the natural islet microenvironment should be mimicked for optimum viability and function, we set out to isolate extracellular matrix components from human pancreas, not only for analytical purposes, but also to be used as supplementary components of encapsulating materials. A protocol was designed to routinely culture different pancreatic tissues (islets, parenchyma and ducts) in the presence of several pancreatic extracellular matrix components and peptide growth factors to enrich the beta cell population in vitro before transplantation into patients. In addition to representing a therapeutic promise, this initiative is an example of productive partnership between the medical and scientific sectors of the university and private enterprises. PMID- 11378657 TI - Development of new heparin-like compounds and other antithrombotic drugs and their interaction with vascular endothelial cells. AB - The anticlotting and antithrombotic activities of heparin, heparan sulfate, low molecular weight heparins, heparin and heparin-like compounds from various sources used in clinical practice or under development are briefly reviewed. Heparin isolated from shrimp mimics the pharmacological activities of low molecular weight heparins. A heparan sulfate from Artemia franciscana and a dermatan sulfate from tuna fish show a potent heparin cofactor II activity. A heparan sulfate derived from bovine pancreas has a potent antithrombotic activity in an arterial and venous thrombosis model with a negligible activity upon the serine proteases of the coagulation cascade. It is suggested that the antithrombotic activity of heparin and other antithrombotic agents is due at least in part to their action on endothelial cells stimulating the synthesis of an antithrombotic heparan sulfate. PMID- 11378658 TI - P-selectin, carcinoma metastasis and heparin: novel mechanistic connections with therapeutic implications. AB - Metastasis is a multistep cascade initiated when malignant cells penetrate the tissue surrounding the primary tumor and enter the bloodstream. Classic studies indicated that blood platelets form complexes around tumor cells in the circulation and facilitate metastases. In other work, the anticoagulant drug heparin diminished metastasis in murine models, as well is in preliminary human studies. However, attempts to follow up the latter observation using vitamin K antagonists failed, indicating that the primary mechanism of heparin action was unrelated to its anticoagulant properties. Other studies showed that the overexpression of sialylated fucosylated glycans in human carcinomas is associated with a poor prognosis. We have now brought all these observations together into one mechanistic explanation, which has therapeutic implications. Carcinoma cells expressing sialylated fucosylated mucins can interact with platelets, leukocytes and endothelium via the selectin family of cell adhesion molecules. The initial organ colonization of intravenously injected carcinoma cells is attenuated in P-selectin-deficient mice, in mice receiving tumor cells pretreated with O-sialoglycoprotease (to selectively remove mucins from cell surfaces), or in mice receiving a single dose of heparin prior to tumor cell injection. In each case, we found that formation of a platelet coating on cancer cells was impeded, allowing increased access of leukocytes to the tumor cells. Several weeks later, all animals showed a decrease in the extent of established metastasis, indicating a long-lasting effect of the short-term intervention. The absence of obvious synergism amongst the three treatments suggests that they all act via a common pathway. Thus, a major mechanism of heparin action in cancer may be inhibition of P-selectin-mediated platelet coating of tumor cells during the initial phase of the metastatic process. We therefore suggest that heparin use in cancer be re-explored, specifically during the time interval between initial visualization of a primary tumor until just after definitive surgical removal. PMID- 11378659 TI - Influence of season and pollution on the antioxidant defenses of the cichlid fish acara (Geophagus brasiliensis). AB - The livers of Geophagus brasiliensis collected from both a non-polluted site and a polluted site were analyzed for different antioxidant defenses, O2 consumption, thiobarbituric acid-reactive substance (TBARS) levels, and histological damage. Compared to controls (116.6 +/- 26.1 nmol g-1), TBARS levels were enhanced at the polluted site (284.2 +/- 25.6 nmol g-1), as also was oxygen consumption (86.6 +/- 11.3 and 128.5 +/- 9.8 micromol O2 min-1 g-1, respectively). With respect to enzymatic antioxidants, increased catalase activities (8.7 +/- 1.3 and 29.2 +/- 2.4 mmol min-1 g-1, respectively), unchanged superoxide dismutase activities (767.2 +/- 113.3 and 563.3 +/- 70.2 U g-1, respectively), and diminished glutathione S-transferase activities (29.0 +/- 3.2 and 14.9 +/- 3.2 micromol min 1 g-1, respectively) were detected. Reduced glutathione (1.91 +/- 0.17 and 1.37 +/- 0.25 mM, respectively), oxidized glutathione (1.50 +/- 0.20 and 0.73 +/- 0.17 mM, respectively), and total glutathione (3.40 +/- 0.26 and 2.07 +/- 0.27 mM, respectively) concentrations were also below control values at the polluted site. Nevertheless, the observed ethoxyresorufin-O-deethylase activities (1.34 +/- 0.11 and 16.7 +/- 0.21 pmol min-1 mg-1, respectively) showed enhanced values at the polluted site. The main histological damage observed in the hepatocytes from fish collected at the polluted site was characterized by heavy lipid infiltration. Fish collected at the end of spring showed higher O2 consumption, higher superoxide dismutase and glutathione S-transferase activities, and higher total and oxidized glutathione concentrations compared to the beginning of autumn. No seasonal changes were observed in catalase activities, glutathione or TBARS levels. Fish chronically exposed to relatively high pollution levels seem to be unable to set up adequate antioxidant defenses, probably due to severe injury to their hepatocytes. The higher antioxidant defenses found at the end of spring are probably related to the enhanced activities during high temperature periods in thermoconforming organisms. PMID- 11378660 TI - Low frequency of p53 mutations in cervical carcinomas among Brazilian women. AB - Human papillomavirus (HPV) infections of the high-risk types are strongly linked to the development of cervical carcinoma. The HPV oncoproteins E6 and E7 are thought to play a crucial role in this process through their interactions with the p53 protein and the retinoblastoma susceptibility gene product pRb, respectively. E6 binds to p53 protein promoting its degradation. This is considered to contribute to the oncogenesis of HPV-associated anogenital cancer. On the other hand, in HPV-negative cervical carcinoma, p53 mutations are thought to have a role in the transformation process. A total of 122 HPV-positive cervical carcinoma tissue samples were evaluated for the presence of mutations in exons 5-8 of the p53 gene by single-stranded conformation polymorphism analysis and DNA sequencing. Only four missense point mutations were detected. These findings suggest that other mechanisms independent of p53 inactivation may play a role in the genesis of cervical carcinomas. PMID- 11378661 TI - Acute promyelocytic leukemia: the study of t(15;17) translocation by fluorescent in situ hybridization, reverse transcriptase-polymerase chain reaction and cytogenetic techniques. AB - Acute promyelocytic leukemia (AML M3) is a well-defined subtype of leukemia with specific and peculiar characteristics. Immediate identification of t(15;17) or the PML/RARA gene rearrangement is fundamental for treatment. The objective of the present study was to compare fluorescent in situ hybridization (FISH), reverse transcriptase-polymerase chain reaction (RT-PCR) and karyotyping in 18 samples (12 at diagnosis and 6 after treatment) from 13 AML M3 patients. Bone marrow samples were submitted to karyotype G-banding, FISH and RT-PCR. At diagnosis, cytogenetics was successful in 10 of 12 samples, 8 with t(15;17) and 2 without. FISH was positive in 11/12 cases (one had no cells for analysis) and positivity varied from 25 to 93% (mean: 56%). RT-PCR was done in 6/12 cases and all were positive. Four of 8 patients with t(15;17) presented positive RT-PCR as well as 2 without metaphases. The lack of RT-PCR results in the other samples was due to poor quality RNA. When the three tests were compared at diagnosis, karyotyping presented the translocation in 80% of the tested samples while FISH and RT-PCR showed the PML/RARA rearrangement in 100% of them. Of 6 samples evaluated after treatment, 3 showed a normal karyotype, 1 persistence of an abnormal clone and 2 no metaphases. FISH was negative in 4 samples studied and 2 had no material for analysis. RT-PCR was positive in 4 (2 of which showed negative FISH, indicating residual disease) and negative in 2. When the three tests were compared after treatment, they showed concordance in 2 of 6 samples or, when there were not enough cells for all tests, concordance between karyotype and RT-PCR in one. At remission, RT-PCR was the most sensitive test in detecting residual disease, as expected (positive in 4/6 samples). An incidence of about 40% of 5' breaks and 60% of 3' breaks, i.e., bcr3 and bcr1/bcr2, respectively, was observed. PMID- 11378662 TI - Scintigraphy and Doppler ultrasonography for the evaluation of obstructive urinary calculi. AB - Forty-seven patients with unilateral obstructive calculi (12 males and 35 females) were submitted to 99mTc-diethylene triamine pentaacetic acid (DTPA) or 99mTc-dimercaptosuccinic acid (DMSA) scans for assessment of renal function. The scans revealed unilateral functional deficit in 68 and 66% of the patients, respectively. A calculus size of 1.1 to 2.0 cm was significantly associated with deficit detected by DTPA, but duration of obstruction and calculus localization were not. After relief of the obstruction, the mean percent renal function of the affected kidney was found to be significantly increased from 25 +/- 12% to 29 +/- 12% in DTPA and from 21 +/- 15% to 24 +/- 12% in DMSA. Initial Doppler ultrasonography performed in 35 patients detected an increased resistive index in 10 (29%). In the remaining patients with a normal resistive index, ureteral urinary jet was observed, indicating partial obstruction. The high frequency of renal function impairment detected by DTPA and of tubulointerstitial damage detected by DMSA as well as the slight amelioration of unilateral renal function after relief of obstruction suggest that scintigraphy assessment may help evaluate the unilateral percentage of renal function and monitor renal function recovery when it occurs. The presence of a urinary jet detected by Doppler ultrasonography further indicates the severity of obstruction and the recovery prognosis. PMID- 11378663 TI - Eradication of Helicobacter pylori infection in patients with duodenal ulcer and non-ulcer dyspepsia and analysis of one-year reinfection rates. AB - Helicobacter pylori (HP) infection is endemic worldwide. The proposed treatment is expensive and there are few reports regarding reinfection rates in Brazil. The aim of this study was to compare the eradication rates obtained with two therapeutic options and to evaluate reinfection one year after treatment. This was a prospective randomized trial with 55 patients. Thirty-nine patients had active duodenal ulcer (DU) and 16 non-ulcer dyspepsia (NUD), and all tested positive for HP. Diagnosis was based on at least two positive tests: ultrarapid urease test, histology and/or culture. Patients were randomized to two groups: group OMC treated with 40 mg omeprazole (once a day), 500 mg metronidazole and 250 mg clarithromycin (twice daily) for 7 days, or group NA treated with 300 mg nizatidine (once a day) and 1000 mg amoxicillin (twice daily) for 14 days. Those patients in whom HP was eradicated were followed up for one year to evaluate reinfection. Twenty-five patients were randomized for OMC and 30 for NA. HP eradication occurred in 20/25 patients (80%) treated with OMC and 13/30 (43%) treated with NA (P = 0.01). After reallocation because of initial treatment failure, the overall eradication rate was 44/51 patients (86%). After an average follow-up of one year, we evaluated 34 patients (23 with DU and 11 with NUD). Reinfection occurred in 3/34 patients (7.6%). We conclude that OMC is effective for HP eradication, and that NA should not be used. Reinfection occurs in 7.6% of the patients in the first year after eradication. PMID- 11378664 TI - High prevalence of alpha-thalassemia among individuals with microcytosis and hypochromia without anemia. AB - In order to determine the contribution of alpha-thalassemia to microcytosis and hypochromia, 339 adult outpatients seen at Unicamp University Hospital (with the exception of the Clinical Hematology outpatient clinics), who showed normal hemoglobin (Hb) levels and reduced mean corpuscular volume and mean corpuscular hemoglobin, were analyzed. Ninety-eight were Blacks (28.9%) and 241 were Caucasians (71.1%). In all cases, Hb A2 and F levels were either normal or low. The most common deletional and nondeletional forms of alpha-thalassemia [ alpha3.7, -alpha4.2, --MED, -(alpha)20.5, alphaHphIalpha, alphaNcoIalpha, alphaalphaNcoI and alphaTSAUDI] were investigated by PCR and restriction enzyme analyses. A total of 169 individuals (49.9%) presented alpha-thalassemia: 145 (42.8%) were heterozygous for the -alpha3.7 deletion (-alpha3.7/alphaalpha) and 18 (5.3%) homozygous (-alpha3.7/-alpha3.7), 5 (1.5%) were heterozygous for the nondeletional form alphaHphIalpha (alphaHphIalpha/alphaalpha), and 1 (0.3%) was a --MED carrier (--MED/alphaalpha). Among the Blacks, 56 (57.1%) showed the alpha3.7/alphaalpha genotype, whereas 12 (12.2%) were -alpha3.7/-alpha3.7 and 1 (1.0%) was an alphaHphIalpha carrier; among the Caucasians, 89 (36.9%) were alpha3.7/alphaalpha, 6 (2.5%) had the -alpha3.7/-alpha3.7 genotype, 4 (1.7%) presented the nondeletional form (alphaHphIalpha/alphaalpha), and 1 (0.4%) was a -MED carrier. These results demonstrate that alpha-thalassemia, mainly through the -alpha3.7 deletion, is an important cause of microcytosis and hypochromia in individuals without anemia. These data are of clinical relevance since these hematological alterations are often interpreted as indicators of iron deficiency. PMID- 11378665 TI - Detection of somatic mutations of the PIG-A gene in Brazilian patients with paroxysmal nocturnal hemoglobinuria. AB - Paroxysmal nocturnal hemoglobinuria (PNH) is an acquired clonal syndrome characterized by intravascular hemolysis mediated by complement, thrombotic events and alterations in hematopoiesis. Basically, the molecular events which underlie the complexity of the syndrome consist of the absence of the glycosylphosphatidylinositol (GPI) anchor as a consequence of somatic mutations in the PIG-A gene, located on the X chromosome. The GPI group is responsible for the attachment of many proteins to the cytoplasmic membrane. Two of them, CD55 and CD59, have a major role in the inhibition of the action of complement on the cellular membrane of blood cells. The absence of GPI biosynthesis can lead to PNH. Since mutations in the PIG-A gene are always present in patients with PNH, the aim of this study was to characterize the mutations in the PIG-A gene in Brazilian patients. The analysis of the PIG-A gene was performed using DNA samples derived from bone marrow and peripheral blood. Conformation-sensitive gel electrophoresis was used for screening the mutation and sequencing methods were used to identify the mutations. Molecular analysis permitted the identification of three point mutations in three patients: one G-->A transition in the 5' portion of the second intron, one T-->A substitution in the second base of codon 430 (Leu430-->stop), and one deletion DeltaA in the third base of codon 63. This study represents the first description of mutations in the PIG-A gene in a Brazilian population. PMID- 11378666 TI - In situ variation of cervical mucus pH during exposure to atmospheric air. AB - The objective of the present study was to determine if exposure of cervical mucus to air during specular examination could modify mucus pH. Detection of changes is justified because of their possible interference with sperm-mucus interaction, since an acidic pH is unfavorable to sperm penetration and is associated with infertility due to the cervical factor. Twenty women with good quality mucus were evaluated. pH measurements of ecto- and endocervical mucus were made in situ using a glass electrode after 0-, 5- and 10-min exposure to air. There was a progressive alkalinization of mucus pH. Mean values of ectocervical mucus pH were 6.91, 7.16 and 7.27, while mean values of endocervical mucus pH were 7.09, 7.34 and 7.46 at 0, 5 and 10 min, respectively. Significant differences were found between the mean values obtained at 0 and 5 min, and at 0 and 10 min (P<0.05), whereas the differences in mean values at 5 and 10 min were not significant at either site. We conclude that 5 to 10 min of exposure to atmospheric air affects cervical mucus pH in a significant way. Since tests used to evaluate sperm-mucus interaction generally have not considered this possibility, we suggest that they should be performed immediately after mucus collection in order to avoid misinterpretation of the results. PMID- 11378667 TI - Rat liver responsiveness to gluconeogenic substrates during insulin-induced hypoglycemia. AB - Hepatic responsiveness to gluconeogenic substrates during insulin-induced hypoglycemia was investigated. For this purpose, livers were perfused with a saturating concentration of 2 mM glycerol, 5 mM L-alanine or 5 mM L-glutamine as gluconeogenic substrates. All experiments were performed 1 h after an ip injection of saline (CN group) or 1 IU/kg of insulin (IN group). The IN group showed higher (P<0.05) hepatic glucose production from glycerol, L-alanine and L glutamine and higher (P<0.05) production of L-lactate, pyruvate and urea from L alanine and L-glutamine. In addition, ip injection of 100 mg/kg glycerol, L alanine and L-glutamine promoted glucose recovery. The results indicate that the hepatic capacity to produce glucose from gluconeogenic precursors was increased during insulin-induced hypoglycemia. PMID- 11378668 TI - Expression of CD40 ligand, interferon-gamma and Fas ligand genes in endomyocardial biopsies of human cardiac allografts: correlation with acute rejection. AB - The purpose of the present study was to investigate the expression (mRNA) of CD40 ligand (CD40L), interferon-gamma (IFN-gamma) and Fas ligand (FasL) genes in human cardiac allografts in relation to the occurrence of acute cardiac allograft rejection as well as its possible value in predicting acute rejection. The mRNA levels were determined by a semiquantitative reverse transcriptase-polymerase chain reaction method in 39 samples of endomyocardial biopsies obtained from 10 adult cardiac transplant recipients within the first six months after transplantation. Biopsies with ongoing acute rejection showed significantly higher CD40L, IFN-gamma and FasL mRNA expression than biopsies without rejection. The median values of mRNA expression in biopsies with and without rejection were 0.116 and zero for CD40L (P<0.003), 0.080 and zero for IFN-gamma (P<0.0009), and 0.156 and zero for FasL (P<0.002), respectively. In addition, the levels of IFN gamma mRNA were significantly increased 7 to 15 days before the appearance of histological evidence of rejection (median of 0.086 in pre-rejection biopsies), i.e., they presented a predictive value. This study provides further evidence of heightened expression of immune activation genes during rejection and shows that some of these markers may present predictive value for the occurrence of acute rejection. PMID- 11378669 TI - Effects of lead and/or zinc exposure during the second stage of rapid postnatal brain growth on delta-aminolevulinate dehydratase and negative geotaxis of suckling rats. AB - Lead has been shown to produce cognitive and motor deficits in young rats that could be mediated, at least in part, by inhibition of the zinc-containing heme biosynthetic enzyme delta-aminolevulinate dehydratase (ALA-D). In the present study we investigated the effects of lead and/or zinc treatment during the second stage of rapid postnatal brain development on brain, kidney and blood ALA-D specific activity, as well as the negative geotaxis behavior of rats. Eight-day old Wistar rats were injected intraperitoneally with saline, lead acetate (8 mg/kg) and/or zinc chloride (2 mg/kg) daily for five consecutive days. Twenty four hours after treatment, ALA-D activity was determined in the absence and presence of DL-dithiothreitol (DTT). The negative geotaxis behavior was assessed in 9- to 13-day-old rats. Treatment with lead and/or zinc did not affect body, brain or kidney weights or brain- or kidney-to-body weight ratios of the animals. In spite of the absence of effect of any treatment on ALA-D specific activity in brain, kidney and blood, the reactivation index with DTT was higher in the groups treated with lead or lead + zinc than in the control group, in brain, kidney and blood (mean +/- SEM; brain: 33.33 +/- 4.34, 38.90 +/- 8.24, 13.67 +/- 3.41; kidney: 33.50 +/- 2.97, 37.60 +/- 2.67, 15.80 +/- 2.66; blood: 63.95 +/- 3.73, 56.43 +/- 5.93, 31.07 +/- 4.61, respectively, N = 9-11). The negative geotaxis response behavior was not affected by lead and/or zinc treatment. The results indicate that lead and/or zinc treatment during the second stage of rapid postnatal brain growth affected ALA-D, but zinc was not sufficient to protect the enzyme from the effects of lead in brain, kidney and blood. PMID- 11378670 TI - Effect of the intracerebroventricular administration of GR 113808, a selective 5 HT4 antagonist, on water intake during hyperosmolarity and hypovolemia. AB - We demonstrate here that acute third ventricle injections of GR 113808, a highly selective 5-HT4 antagonist, decrease water intake induced by a previous salt load while potentiating drinking elicited by hypovolemia induced by previous subcutaneous administration of polyethylene glycol in male Wistar rats (200 +/- 20 g). At the dose of 160 nmol/rat, third ventricle injections of GR 113808 induced a significant reduction of water intake in salt-loaded animals after 120 min as compared to salt-loaded animals receiving third ventricle injections of saline (salt load + GR = 3.44 +/- 0.41 ml, N = 12; salt load + saline = 5.74 +/- 0.40 ml, N = 9). At the dose of 80 nmol/rat, GR 113808 significantly enhanced water intake in hypovolemic animals after 120 min as compared to hypovolemic animals receiving third ventricle injections of saline (hypovol + GR = 4.01 +/- 0.27 ml, N = 8; hypovol + saline = 2.41 +/- 0.23 ml, N = 12). We suggest that central 5-HT4 receptors may exert a positive drive on water intake due to hyperosmolarity and a negative input on drinking provoked by hypovolemia. PMID- 11378671 TI - Behavioral and electroencephalographic analysis of seizures induced by intrahippocampal injection of granulitoxin, a neurotoxic peptide from the sea anemone Bunodosoma granulifera. AB - In this study, the behavioral and electroencephalographic (EEG) analysis of seizures induced by the intrahippocampal injection in rats of granulitoxin, a neurotoxic peptide from the sea anemone Bunodosoma granulifera, was determined. The first alterations occurred during microinjection of granulitoxin (8 microg) into the dorsal hippocampus and consisted of seizure activity that began in the hippocampus and spread rapidly to the occipital cortex. This activity lasted 20 30 s, and during this period the rats presented immobility. During the first 40 50 min after its administration, three to four other similar short EEG seizure periods occurred and the rats presented the following behavioral alterations: akinesia, facial automatisms, head tremor, salivation, rearing, jumping, barrel rolling, wet dog shakes and forelimb clonic movements. Within 40-50 min, the status epilepticus was established and lasted 8-12 h. These results are similar to those observed in the acute phase of the pilocarpine model of temporal lobe epilepsy and suggest that granulitoxin may be a useful tool not only to study the sodium channels, but also to develop a new experimental model of status epilepticus. PMID- 11378672 TI - The early facilitatory effect of a peripheral spatially noninformative prime stimulus depends on target stimulus features. AB - We investigated the dependency of the early facilitatory effect of a prime stimulus (S1) on the physical characteristics of the target stimulus (S2). A go no go reaction time paradigm was used. The S1 was a gray ring and the S2s were a white vertical line, a white horizontal line, a white cross and a white small ring, all inside a white ring with the same dimensions as the S1. S1 onset-S2 onset asynchrony was 100 ms. The stimuli appeared randomly in any one of the quadrants of a monitor screen. The S2 could occur at the same position as the S1 or at a different one. We observed a strong facilitatory effect when the vertical line or the horizontal line was the go stimulus and no effect when the cross was the go stimulus. These results show that the features of the target stimulus can be decisive for the appearance of the facilitatory effect of a peripheral spatially noninformative prime stimulus. PMID- 11378673 TI - Central injection of captopril inhibits the blood pressure response to intracerebroventricular choline. AB - In the present study, we investigated the involvement of the brain renin angiotensin system in the effects of central cholinergic stimulation on blood pressure in conscious, freely moving normotensive rats. In the first step, we determined the effects of intracerebroventricular (icv) choline (50, 100 and 150 microg) on blood pressure. Choline increased blood pressure in a dose-dependent manner. In order to investigate the effects of brain renin-angiotensin system blockade on blood pressure increase induced by choline (150 microg, icv), an angiotensin-converting enzyme inhibitor, captopril (25 and 50 microg, icv), was administered 3 min before choline. Twenty-five microg captopril did not block the pressor effect of choline, while 50 microg captopril blocked it significantly. Our results suggest that the central renin-angiotensin system may participate in the increase in blood pressure induced by icv choline in normotensive rats. PMID- 11378674 TI - Comparison of the quantification of caffeine in human plasma by gas chromatography and ELISA. AB - In the present study we evaluated the precision of the ELISA method to quantify caffeine in human plasma and compared the results with those obtained by gas chromatography. A total of 58 samples were analyzed by gas chromatography using a nitrogen-phosphorus detector and routine techniques. For the ELISA test, the samples were diluted to obtain a concentration corresponding to 50% of the absorbance of the standard curve. To determine whether the proximity between the I50 of the standard curve and that of the sample would bring about a more precise result, the samples were divided into three blocks according to the criterion of difference, in modulus, of the I50 of the standard curve and of the I50 of the sample. The samples were classified into three groups. The first was composed of 20 samples with I50 up to 1.5 ng/ml, the second consisted of 21 samples with I50 ranging from 1.51 to 3 ng/ml, and the third of 17 samples with I50 ranging from 3.01 to 13 ng/ml. The determination coefficient (R2 = 0.999) showed that the data obtained by gas chromatography represented a reliable basis. The results obtained by ELISA were also reliable, with an estimated Pearson correlation coefficient of 0.82 between the two methods. This coefficient for the different groups (0.88, 0.79 and 0.49 for groups 1, 2 and 3, respectively) showed greater reliability for the test with dilutions closer to I50. PMID- 11378675 TI - Effects of L-arginine on the diaphragm muscle twitches elicited at different frequencies of nerve stimulation. AB - In rats, the nitric oxide (NO)-synthase pathway is present in skeletal muscle, vascular smooth muscle, and motor nerve terminals. Effects of NO were previously studied in rat neuromuscular preparations receiving low (0.2 Hz) or high (200 Hz) frequencies of stimulation. The latter frequency has always induced tetanic fade. However, in these previous studies we did not determine whether NO facilitates or impairs the neuromuscular transmission in preparations indirectly stimulated at frequencies which facilitate neuromuscular transmission. Thus, the present study was carried out to examine the effects of NO in rat neuromuscular preparations indirectly stimulated at 5 and 50 Hz. The amplitude of muscular contraction observed at the end (B) of a 10-s stimulation was taken as the ratio (R) of that obtained at the start (A) (R = B/A). S-nitroso-N-acetylpenicillamine (200 microM), superoxide dismutase (78 U/ml) and L-arginine (4.7 mM), but not D arginine (4.7-9.4 mM), produced an increase in R (facilitation of neurotransmission) at 5 Hz. However, reduction in the R value (fade of transmission) was observed at 50 Hz. N G-nitro-L-arginine (8.0 mM) antagonized both the facilitatory and inhibitory effects of L-arginine (4.7 mM). The results suggest that NO may modulate the release of acetylcholine by motor nerve terminals. PMID- 11378676 TI - Kidney function after left renal vein ligation in the dog. AB - The ligature of the left renal vein is an alternative whenever this vessel is injured. The purpose of this study was to evaluate the capacity of the affluents of the left renal vein, proximal to the ligature, to maintain tissue vitality and function of the left kidney. Fifteen mongrel male dogs were divided in 3 groups of 5 dogs: Group I (control) - a laparotomy was performed, and the abdominal structures were only identified; Group II - the left renal vein was tied, close to vena cava; Group III - the same procedure as for Group II and a right nephrectomy. Blood urea nitrogen and serum creatinine levels were measured before the procedure, and every 3 days during 4 weeks in the postoperative period. Renal arteriography and an excretory urogram were performed on the animals that survived 60 days. Thereafter, or immediately after precocious death, the kidneys were removed for histological examination. All the animals of Group III died before two months (mean = 10.5 +-3.2 days), while the animals of Group II survived during that period. There was a complete exclusion of the left kidney in all dogs that underwent renal vein ligature. In the animals of Group II, the renal cortico-medullary limits could not be identified. At microscopy, the aspect was suggestive of nephrosclerosis. In the animals of Group III, the left kidney was enlarged, and a great amount of intravascular and intrapelvic blood clots were observed. At microscopy, extensive areas of necrosis, inflammatory infiltration, and hemorrhage were identified. In conclusion, the tributaries of the renal vein were not sufficient to maintain the tissue vitality and function of the left kidney after ligature of its main vein. PMID- 11378677 TI - Pain evaluation of patients with fibromyalgia, osteoarthritis, and low back pain. AB - The purpose of this study was to evaluate and compare pain as reported by outpatients with fibromyalgia, osteoarthritis, and low back pain, in view of designing more adequate physical therapy treatment. PATIENTS AND METHODS: A Portuguese version of the McGill Pain Questionnaire - where subjects are asked to choose, from lists of pre-categorized words, one or none that best describes what they feel - was used to assess pain intensity and quality of 64 patients, of which 24 had fibromyalgia, 22 had osteoarthritis, and 18 had low back pain. The pre-categorized words were organized into 4 major classes - sensory, affective, evaluative, and miscellaneous. RESULTS: Patients with fibromyalgia reported, comparatively, more intense pain through their choice of pain descriptors, both sensory and affective; they also chose a higher number of words from these classes than patients in the other groups and were the only ones to choose specific affective descriptors such as "vicious", "wretched", "exhausting", "blinding". CONCLUSION: Assuming that each disease presents unique qualities of pain experience, and that these can be pointed out by means of this questionnaire by patients' choice of specific groups of words, the findings suggest that fibromyalgia include not only a physical component, but also a psycho-emotional component, indicating that they require both emotional/affective and physical care. PMID- 11378678 TI - Factors affecting Helicobacter pylori eradication using a seven-day triple therapy with a proton pump inhibitor, tinidazole and clarithromycin, in Brazilian patients with peptic ulcer. AB - Triple therapy is accepted as the treatment of choice for H. pylori eradication. In industrialized countries, a proton pump inhibitor plus clarithromycin and amoxicillin or nitroimidazole have shown the best results. Our aims were: 1. To study the eradication rate of the association of a proton pump inhibitor plus tinidazole and clarithromycin on H. pylori infection in our population. 2. To determine if previous treatments, gender, age, tobacco, alcohol use, and non steroidal anti-inflammatory drugs (NSAIDs) change the response to therapy. METHODS: Two hundred patients with peptic ulcer (upper endoscopy) and H. pylori infection (histology and rapid urease test - RUT) were included. A proton pump inhibitor (lanzoprazole 30 mg or omeprazole 20 mg), tinidazole 500 mg, and clarithromycin 250 mg were dispensed twice a day for a seven-day period. Eradication was assessed after 10 to 12 weeks of treatment through histology and RUT. RESULTS: The eradication rate of H. pylori per protocol was 65% (128/196 patients). This rate was 53% for previously treated patients, rising to 76% for not previously treated patients, with a statistical difference p<0.01. No significant difference was observed regarding sex, tobacco use, alcohol consumption, and NSAID use, but for elderly patients the difference was p = 0.05. Adherence to treatment was good, and side effects were mild. CONCLUSIONS: A proton pump inhibitor, tinidazole, and clarithromycin bid for seven days resulted in H. pylori eradication in 65% of the patients. Previous treatments were the main cause of treatment failure. PMID- 11378679 TI - Monitoring the treatment of sepsis with vancomycin in term newborn infants. AB - A prospective study was conducted to determine if standardized vancomycin doses could produce adequate serum concentrations in 25 term newborn infants with sepsis. PURPOSE: The therapeutic response of neonatal sepsis by Staphylococcus sp. treated with vancomycin was evaluated through serum concentrations of vancomycin, serum bactericidal titers (SBT), and minimum inhibitory concentration (MIC). METHOD: Vancomycin serum concentrations were determined by the fluorescence polarization immunoassay technique, SBT by the macro-broth dilution method, and MIC by diffusion test in agar. RESULTS: Thirteen newborn infants (59.1%) had adequate peak vancomycin serum concentrations (20 - 40 mg/mL) and one had peak concentration with potential ototoxicity risk (>40 microg/mL). Only 48% had adequate trough concentrations (5 - 10 mg/mL), and seven (28%) had a potential nephrotoxicity risk (>10 microg/mL). There was no significant agreement regarding normality for peak and trough vancomycin method (McNemar test : p = 0.7905). Peak serum vancomycin concentrations were compared with the clinical evaluation (good or bad clinical evolution) of the infants, with no significant difference found (U=51.5; p=0.1947). There was also no significant difference between the patients' trough concentrations and good or bad clinical evolution (U = 77.0; p=0.1710). All Staphylococcus isolates were sensitive to vancomycin according to the MIC. Half of the patients with adequate trough SBT (1/8), also had adequate trough vancomycin concentrations and satisfactory clinical evolution. CONCLUSIONS: Recommended vancomycin schedules for term newborn infants with neonatal sepsis should be based on the weight and postconceptual age only to start antimicrobial therapy. There is no ideal pattern of vancomycin dosing; vancomycin dosages must be individualized. SBT interpretation should be made in conjunction with the patient's clinical presentation and vancomycin serum concentrations. Those laboratory and clinical data favor elucidation of the probable cause of patient's bad evolution, which would facilitate drug adjustment and reduce the risk of toxicity or failing to achieve therapeutic doses. PMID- 11378680 TI - Role of colonoscopy in colorectal cancer. AB - Colorectal cancer (CRC) represents the third most common malignancy throughout the world. Little or no improvement in survival has been effectively achieved in the last 50 years. Extensive epidemiological and genetic data are able to identify more precisely definite risk-groups so screening and early diagnosis can be more frequently accomplished. CRC is best detected by colonoscopy, which allows sampling for histologic diagnosis. Colonoscopy is the gold standard for detection of small and premalignant lesions, although it is not cost-effective for screening average-risk population. Colonoscopic polypectomy and mucosal resection constitute curative treatment for selective cases of invasive CRC. Similarly, alternative trans-colonoscopic treatment can be offered for adequate palliation, thus avoiding surgery. PMID- 11378681 TI - The suitability of hepatocyte culture models to study various aspects of drug metabolism. AB - Since the liver is the main organ involved in the metabolism and the toxicity of xenobiotics, isolated rat hepatocytes have been increasingly used in recent years as a model to identify pharmacological and toxicological responses of drugs. However, it is generally recognised that isolated hepatocytes retain most of their functions only for a short period. For this reason, numerous models and techniques have been developed to study and improve the metabolic capacity of hepatocytes in vitro over an extended time period and in application for drug metabolism studies. In the present study, we compared four different cell culture models to fulfill these requirements and have therefore harvested hepatocytes and cultured them in different culture systems over two weeks. In order to prove certain advantages or disadvantages of each model, we compared the metabolic capacity, albumin secretion, the release of cytosolic and mitochondrial enzymes, as well as the capacity to metabolise diclofenac (DF). We found that rat hepatocytes in all studied culture models (except the Unisyn Bioreactor) were able to metabolise DF to the same extent as found in vivo. However, the concentration of metabolites was found to decrease with culture time using the monolayer although the DF metabolite level in the collagen Sandwich culture was higher than that of the monolayer culture. The 3D-membrane bioreactor preserved the metabolic capacity for a prolonged period of time. The concentrations of DF metabolites in the Unisyn hollow fiber bioreactor were below the detection limit, which corresponded to other parameters such as albumin secretion and cytochrome P450 activity, disqualifying this culture system clearly for the use of in vitro primary hepatocyte cultures. The other three systems all have their place in drug metabolism with different advantages. However, our studies clearly showed that hepatocytes cultured within a collagen sandwich or in the 3D-membrane bioreactor qualify to study various aspects of drug metabolisms over a long time period. Further studies are needed to prove if the later two culture models may really help to reduce animal testing. PMID- 11378682 TI - Reconstructed skin equivalents for assessing percutaneous drug absorption from pharmaceutical formulations. AB - Excised human skin has so far been considered to be one of the most suitable in vitro methods to evaluate the penetration of dermatologically applied substances. The limited supply and the relatively high donor variability stimulated many research groups to use animal skin as a substitute for human skin. Since nowadays reconstructed skin equivalents are commercially available, we examined these cultures for their suitability as a percutaneous absorption model for different pharmaceutical formulations. One such equivalent is EpiDerm (EPI-606, MatTek corporation, Ashland Massachusetts) which was investigated using the lipophilic model drug flufenamic acid. Permeation studies with the Franz diffusion cell were undertaken to evaluate the model for the establishment of a new in vitro method to study the percutaneous absorption of different dosage forms. The drug was applied in two pharmaceutical formulations to the intact surface of the skin disk: dissolved in wool alcohol ointment (0.1125 %), and dissolved in Soerensen phosphate buffer pH 7.4 (0.1125% solution). HPLC was used for the analysis of drug content. It was shown that the model forms a barrier towards diffusion by comparing the permeation across the tissue-free inserts to the equivalents. Flux values were calculated and the permeation across the skin equivalent from the solution was noted to be almost forty times higher than from the ointment. Two different batches of the skin equivalent showed no statistically significant difference. Finally the permeability of the reconstructed skin was compared to human epidermis, and a five times higher flux value was found for the skin equivalent model. Our results suggest that reconstructed skin equivalents based on human keratinocytes have potential as a pharmaceutical test system to study dermal drug transport from topical formulations. PMID- 11378683 TI - [Replacement of the pyrogen-test as batch control test for the biological substances aprotinin and urokinase]. AB - In the Pharmeuropa Vol. 10, No. 4, December 1998, the European Pharmacopoeia Commission published an appeal to replace the pyrogen test by the limulus test for a total of 10 biological substances. These substances included the active ingredients aprotinin and urokinase, contained in our sales products Antagosan and Actosolv. After successful validation of the limulus test, we calculated endotoxin limit concentrations for these and submitted these to the European Department for Quality of Medicines (EDQM). We expect - not least on the basis of the data submitted by ourselves - that the pyrogen test will be replaced in future as a quality control test by the limulus test in testing aprotinin and urokinase compounds for the presence of pyrogenic substances. This leads to a reduction in the number of rabbits used as opposed to the current pyrogen test for testing the stated compounds. PMID- 11378684 TI - Internet laboratory for predicting harmful effects triggered by drugs and chemicals. Concept and call for co-operation. AB - It is our objective to establish a virtual laboratory on the Internet to allow for an in silico estimation of harmful effects triggered by drugs, chemicals and their metabolites. Presently, our database includes validated models for five biological targets -- the Aryl hydrocarbon, serotonin 5HT-2A, cannabinoid, GABA (gamma-amino butter acid), and steroid receptors. It shall be continuously extended to include surrogates for any bioregulator known or presumed to mediate harmful effects. Free access to this virtual laboratory shall allow any interested party to estimate the harmful potential of a given substance prior to its synthesis. This is achieved by generating the three-dimensional structure of the compound and its possible metabolites in the computer, followed by calculating their binding affinity towards each receptor surrogate in the database. Only compounds/metabolites passing through this surrogate battery without displaying a significant affinity towards any member may be cleared for synthesis and preclinical studies. This way, potentially harmful compounds can be withdrawn from the evaluation pipeline before in vivo test are conducted, hence contributing to the reduction of animal testing in chemical and pharmaceutical research and development. PMID- 11378685 TI - [Report on the ICCVAM workshop on in vitro methods for assessing acute systemic toxicity]. AB - It was suggested in the ICCVAM workshop that the Register of Cytotoxicity (RC), using in vitro cytotoxicity data to predict the in vivo starting doses, should be implemented into acute toxicity testing as soon as possible. The validity of the in vitro cytotoxicity data to establish appropriate starting doses for acute toxicity testing will be assessed experimentally. Secondly, in order to replace the use of animals in acute lethality testing a formal validation will be conducted in which the ability to predict rodent LD50 values and toxicity classes from cytotoxicity data will be evaluated. PMID- 11378686 TI - [In vitro methods for phototoxicity and photocarcinogenicity testing of drugs]. AB - Phototoxicity is an acknowledged property of some UV and/or visible light absorbing substances some of which are used as pharmaceuticals or in cosmetic preparations. In recent years attention has been called upon the fact that toxic intermediates that are generated upon photoactivation of a substance can also lead to DNA damage. Such damage may lead to mutated/initiated skin cells which in turn can contribute to an elevated skin cancer risk. The method of choice to test for photo-related skin carcinogenesis is a 1-year study in genetically hairless mice in which the formation of skin papilloma and their latency time are assessed. Here, in vitro test approaches to test for photogenotoxicity can be used in a tiered assessment approach asking the use of in vitro genotoxicity tests for prediction of rodent/human carcinogenicity. In the past few years some effort has been put into the evaluation for such systems, in particular standard test protocols have been generated for the in vitro photo-micronucleus test and the in vitro photo-comet assay with Chinese hamster V79 cells. The data that have been produced so far show promising results regarding the implementation of these systems in a tiered approach for photocarcinogenicity assessment of UV- and/or visible light absorbing substances but the systems will have to be validated in further collaborative studies. PMID- 11378687 TI - Prediction of embryotoxic effects of valproic acid-derivatives with molecular in vitro methods. AB - Therapy with the antiepileptic drug valproic acid (2-propylpentanoic acid, VPA) during early pregnancy can cause similar teratogenic effects (neural tube defects) in human and mice. In this study a new molecular bioassay is presented using following endpoints: differentiation of F9 teratocarcinoma cells, altered cell morphology, induction of possible targeted genes, and the induction of viral RSV-promoter. The induction of a transiently transfected viral (RSV) promoter driven luciferase gene by VPA was used to screen a set of VPA-derivatives. Structure-activity investigations showed: the longer the aliphatic side chain the more the induction of the RSV-reporter gene. The specific induction was stereoseletive. The teratogenic enantiomer S-4-yn-VPA (2-propyl-4-pentynoic acid) induced the RSV-driven reporter gene while the non teratogenic R-4-yn-VPA does not. Heptyl-4-yn-VPA was the most potent teratogen in vitro and in vivo. Non teratogenic VPA-derivatives like R-4-yn-VPA and 2-en-VPA (2-propyl-2-pentenoic acid) were ineffective in this system. Thus, the teratogenic effect of VPA and VPA-derivatives in the mouse correlated with the specific induction of the viral RSV-promoter controlled reporter in F9-cells. Acid compounds such as fatty acids are known to interact with peroxisome proliferator-activated receptors (PPARs). To test structure-activity relationships by VPA or its derivatives we used CHO cells stably expressing hybrid proteins of the ligand-binding domain of either of the PPARs. The teratogen VPA and the teratogenic derivatives of VPA activated the PPAR-delta construct in a very specific structure- and stereoselective way which correlated well with the activities in the reporter gene assay (bioassay) and those in vivo. No such correlation was found with respect to activation of PPAR alpha or PPAR-gamma. These structure-activity relationships indicate that PPAR delta may be a potential mediator of VPA-induced differentiation of F9 cells and may possibly be involved in the mechanism of teratogenicity of VPA in vivo. Furthermore two bioassays were designed with clearly defined endoints, amenable to automation and screening of great number of compounds. The test system allows to replace animal experiments in the preclinical development of new antiepileptics drugs with reduced teratogenic risk. Supported by BgVV-ZEBET (Berlin). PMID- 11378688 TI - [Permanent embryonic mouse germ cell-lines, an in vitro alternative to in vivo germ cell mutagenicity tests]. AB - Germ cell mutagenesis is required by the 7th amendment of the directive 67/548 EEC into the national regulations on existing chemicals. Officially accepted in vivo test systems for stage specific mutagenicity are the dominant lethal (DL) test and the specific locus test (SLT) in mice. An acceptable in vitro alternative designed to address germ cell mutagenesis and discriminate between male and female specific effects is not available at present. In order to offer a sensitive and predictive in vitro method to assess the genotoxic potential of chemical agents on male and female reproduction, we established primordial germ (PG) cell-derived permanent embryonic germ (EG) cell lines of the mouse (strain BALB/cJ). The differences in developmental sensitivity of the EG(3) cell line and differentiated fibroblast cells 3T3 were comparatively tested with cytotoxicity assay (MTT test ) and genotoxic studies (SCE-assay) under identical test conditions. The concentration-response curves reflected the female cell line EG(3) to be extremely sensitive concerning cytotoxic and genotoxic endpoints. Therefore this cell line was used to classify in vivo genotoxic and non-genotoxic test substances with different potential endpoints. Applying linear discriminant analysis three endpoints were identified for the correct classification (100%) of all test chemicals, namely the SCE(200) value (increase of 200% in the mean number of SCEs per metaphase spread) for EG(3) (3 hrs and 24 hrs assay) and the IC(5)0 value for EG(3) after 3 hrs of exposure to test chemicals. PMID- 11378689 TI - [White Paper of the Commission of European Community--strategy for a future chemicals policy]. PMID- 11378803 TI - Systematic review concerning the age of introduction of complementary foods to the healthy full-term infant. AB - OBJECTIVES: To evaluate existing data from published studies investigating outcomes (growth, nutritional status and morbidity) in relation to the age of introduction of complementary foods to the healthy full-term infant. To assess the methodological quality of current scientific evidence on which infant feeding recommendations are based. SEARCH STRATEGY: A database was created following a search of electronic databases MEDLINE and BIDS, a handsearch of relevant journals and consultation with international experts in infant feeding practices. The search was conducted during a 6-month period and encompassed the years 1982 1998. INCLUSION CRITERIA: Randomised/non-randomised controlled trials and cohort studies investigating the relationship to the health of full-term infants of the introduction of complementary foods to childhood health. DATA COLLECTION AND ANALYSIS: Assessment of studies for inclusion and their compliance with methodological criteria was undertaken independently by two reviewers and discrepancies were resolved by discussion. RESULTS: From over 400 published papers identified, 33 met specific inclusion criteria. Significant differences in baseline characteristics of subjects in included studies meant that data were not amenable to meta-analysis. The consensus of the authors was that, of the 33 included publications, 13 contained data supporting the current WHO recommendations which attempt to accommodate all infants, including those whose nutritional requirements are not met by exclusive breast-feeding for 6 months. An equal number contained data that would support a recommendation for delaying the introduction of complementary foods until 6 months of age within the study population. The remaining seven were unable to provide evidence to support a change from the current WHO infant feeding recommendations which state that 'infants should be fed exclusively on breast milk from birth to 4-6 months of age'. None of the studies met all the methodological criteria. CONCLUSIONS: This review has shown that there is a lack of clear evidence to either support or refute a change to the current recommendations for the age of introduction of complementary foods to the breast milk or formula fed infant. Whilst exclusive breast feeding for the first 6 months of life can support growth and development in some infants, sub-groups have been identified within certain populations who may require complementary feeding prior to this age. PMID- 11378804 TI - The metabolism of linoleic acid in healthy subjects after intake of a single dose of (13)C-linoleic acid. AB - OBJECTIVE: To investigate the in vivo oxidation of (13)C18:2n-6 and its conversion into longer-chain polyunsaturates (LCPs) in healthy subjects. DESIGN: Blood samples were collected from six subjects before (fasted) and 7, 11 (non fasted), 24, 48, 72, 168 and 336 h (fasted) after ingestion of a single bolus of 45 mg uniformly labeled (13)C18:2n-6 dissolved in 8 g olive oil. In three subjects, breath was also sampled and CO(2) production measured every hour during the first 12 h. Subjects consumed their habitual diets. Plasma (13)C-enrichments were measured by GC-C-IRMS and fatty acid compositions by GC/FID. SETTING: Maastricht University, Department of Human Biology. SUBJECTS: Three men and three women, recruited by local advertisement. RESULTS: The tracer/tracee ratio (TTR) of C18:2n-6 in plasma total lipids was already increased 5 h after tracer intake. The mean peak amount (+/-s.e.m) of (13)C18:2n-6 (3.4+/-0.8 mg; 7.6% of dose) was found after about 17 h, (13)C18:3n-6 (0.018+/-0.008 mg; 0.04% of dose) after 7-48 h, and (13)C20:3n-6 (0.028+/-0.011 mg; 0.06% of dose) after 48-336 h. Time to peak TTRs of C20:4n-6 varied between subjects and were on average 0.022+/-0.006 mg (0.05% of dose). The proportion of (13)C18:2n-6 recovered in breath after 12 h ranged between 16.8 and 25.1%. CONCLUSIONS: These findings suggest that a single bolus of 45 mg U-(13)C18:2n-6 can be used to study the oxidation of (13)C18:2n-6. However, because of the low TTRs for C20:4n-6, a higher dose is recommended for studying the conversion of (13)C18:2n-6 into LCPs. In addition, since only about 35% of the tracer was found in plasma total lipids and as (13)CO(2) in breath, it might be necessary to study other accessible lipid fractions as well to study the overall conversion of linoleic acid. PMID- 11378805 TI - Mechanisms of action of beta-glucan in postprandial glucose metabolism in healthy men. AB - OBJECTIVE: To assess whether beta-glucan (which is fermented in the colon) lowers postprandial glucose concentrations through mechanisms distinct from a delayed carbohydrate absorption and inhibits de novo lipogenesis. DESIGN: Administration of frequent small meals each hour over 9 h allows a rate of intestinal absorption to be reached which is independent of a delayed absorption. A group of 10 healthy men received either an isoenergetic diet containing 8.9 g/day beta-glucan or without beta-glucan for 3 days. On the third day, the diet was administered as fractioned meals ingested every hour for 9 h. SETTING: Laboratory for human metabolic investigations. SUBJECTS: Ten healthy male volunteers. MAIN OUTCOME MEASURES: Plasma glucose and insulin concentrations, glucose kinetics, glucose oxidation, de novo lipogenesis. RESULTS: On the third day, plasma glucose and free fatty acid concentrations, carbohydrate and lipid oxidation, and energy expenditure were identical with beta-glucan and cellulose. Plasma insulin concentrations were, however, 26% lower with beta-glucan during the last 2 h of the 9 h meal ingestion. Glucose rate of appearance at steady state was 12% lower with beta-glucan. This corresponded to a 21% reduction in the systemic appearance rate of exogenous carbohydrate with beta-glucan, while endogenous glucose production was similar with both diets. De novo lipogenesis was similar with and without beta-glucan. CONCLUSION: Administration of frequent meals with or without beta-glucan results in similar carbohydrate and lipid metabolism. This suggests that the lowered postprandial glucose concentrations which are observed after ingestion of a single meal containing beta-glucan are essentially due to a delayed and somewhat reduced carbohydrate absorption from the gut and do not result from the effects of fermentation products in the colon. PMID- 11378806 TI - Effect of phenol-rich extra virgin olive oil on markers of oxidation in healthy volunteers. AB - OBJECTIVE: We studied whether consumption of phenol-rich extra virgin olive oil affects the susceptibility of low density lipoproteins (LDL) to oxidation and other markers of oxidation in humans. DESIGN: Randomized cross-over intervention trial, stratified according to sex, age and energy intake. SETTING: Division of Human Nutrition and Epidemiology, Wageningen University, The Netherlands. SUBJECTS: Forty-six healthy men and women completed the study. INTERVENTION: Subjects consumed two diets supplying 69 g per day of extra virgin olive oil either rich or poor in phenols for 3 weeks each. The mean difference in phenol intake between the treatments was 18 mg per day. Vitamin E intake was low during the whole study. Fasting blood samples were taken twice at the end of each period. RESULTS: Resistance of LDL and high density lipoprotein (HDL) to oxidation was not affected by treatment. The mean lag time of copper-induced formation of conjugated dienes was 1.6 min shorter in LDL and 0.4 min longer in HDL after the high phenol diet. Other markers of antioxidant capacity in plasma were also not affected: mean lipid hydroperoxides were 0.07 micromol/l higher, mean malondialdehydes were 0.001 micromol/l higher, mean protein carbonyls were 0.001 nmol/mg protein lower, and the mean ferric reducing ability of plasma (FRAP) was 0.006 mmol/l higher after the high phenol diet. All 95% confidence intervals enclosed zero. Serum cholesterol concentrations were not affected by the treatment. CONCLUSION: Consumption of 18 mg per day of phenols from extra virgin olive oil for 3 weeks did not affect LDL or HDL oxidation or other markers of antioxidant capacity in fasting plasma samples. PMID- 11378807 TI - Fruit and vegetable intake and population glycosylated haemoglobin levels: the EPIC-Norfolk Study. AB - OBJECTIVE: To investigate whether self-reported frequency of fruit and vegetable consumption was associated with HbA(1C) levels in individuals not known to have diabetes, and what dietary and lifestyle factors might explain this association. DESIGN: Cross-sectional study. SETTING: The EPIC-Norfolk Study, a population based cohort study of diet and chronic disease. SUBJECTS AND METHODS: A total of 2678 men and 3318 women (45-74 y) not known to have diabetes reported weekly consumption of fruit, green leafy vegetables and other vegetables. RESULTS: Among men, 274 (10.2%) reported seldom or never eating fruit and 127 (4.7%) seldom or never eating green leafy vegetables. Corresponding numbers in women were 157 (4.7%) and 92 (2.8%), respectively. Participants who reported never or seldom having both fruit and green leafy vegetables had higher mean (s.d.) HbA(1C) measurements (5.43% (0.71)) than those who reported more frequent consumption (5.34% (0.67); P=0.046). Differences by category of fruit or green leafy vegetable consumption were not substantially changed after adjustment for saturated fat, dietary fibre and plasma vitamin C. CONCLUSION: These findings support the hypothesis that high intake of fruit and green leafy vegetables may influence glucose metabolism independent of dietary fibre or vitamin C alone and that increased consumption may contribute to the prevention of diabetes. PMID- 11378808 TI - Energy density, energy intake and weight status in a large free-living sample of Chinese adults: exploring the underlying roles of fat, protein, carbohydrate, fiber and water intakes. AB - OBJECTIVE: This paper uses observational data to explore what dietary constituents might be responsible for associations between energy density, energy intake and weight status among free-living individuals. DESIGN: Cross-sectional data on 5783 Chinese adults (aged 20-59 y) from the 1991 China Health and Nutrition Surveys were used to test for associations between energy density, energy intake and weight status, controlling for age, sex, height, activity level, smoking status, urban residence and income. Pearson correlation coefficients were calculated to identify nutrient intake correlates of energy density. Replacing energy density with its nutrient correlates (3 day mean estimates of fat, protein, fiber and water intakes) in the models predicting energy intake and overweight status, the independent effects of specific nutrients were investigated. RESULTS: Energy density was positively and significantly associated with energy intake and overweight status. Energy density was significantly correlated with every nutrient examined, although the correlations were weak for most variables except water intake. Only water intake behaved consistently across analyses. It was negatively correlated with energy density, negatively, significantly and independently associated with energy intake as well as overweight status. Despite positive associations with energy intake, fat and protein intake were not significant predictors of overweight status. Fiber intake was strongly and positively associated with overweight status. CONCLUSIONS: Of the nutrients examined, only water intake appeared to explain the effects of energy density on energy intake and overweight status. PMID- 11378809 TI - Validity and reproducibility of a self-administered questionnaire to determine dietary supplement users among Japanese. AB - OBJECTIVE: To evaluate reproducibility and validity of a self-administered food frequency questionnaire (FFQ) to determine the dietary supplement use. DESIGN: Questionnaire data were compared with dietary records (DR) in four 7 day periods for validity. FFQ1 and FFQ2 administered at an interval of approximately one year were compared for reproducibility. SETTING AND SUBJECTS: A total of 361 samples were selected for a validation study of a questionnaire from the subgroup of the Japan Public Health Center-based prospective Study on cancer and cardiovascular disease (JPHC Study) cohort II. Categories of dietary supplements in the FFQ were 'multivitamins', 'beta-carotene', 'vitamin C', 'vitamin E', and 'other supplements'. For each category, inquiries were made as to the brand name, frequency and duration of use. A dietary supplement user was defined as a subject who used a dietary supplement one or more times a week for a year or longer. RESULTS: Prevalence of overall supplement use was 10.9%. Percentage agreement between FFQ1 and FFQ2 was 91.7%. Sensitivity and specificity of supplement use by FFQ were 80.6% and 89.2%, respectively. Sensitivity was high for 'vitamin C'. When the self-reported categories were corrected, sensitivity for 'multivitamins' and 'vitamin E' improved. CONCLUSIONS: The FFQ on dietary supplements used for the JPHC 5 y follow-up survey was valid to determine overall use and well-defined dietary supplement use such as vitamin C. Categories must be corrected when determining multivitamin and 'vitamin E'. PMID- 11378810 TI - Social position and nutrition: a gradient relationship in Canada and the USA. AB - OBJECTIVE: [corrected] To study the existence of a graded relationship between the nutrient content of the diet and some measures of social position. DESIGN AND SETTING: The graded relationship hypothesis was verified by secondary analysis performed on two different nutrition surveys: the Quebec Nutrition Survey (QNS) and the NHANES III, both based on a single 24 h recall. The data of these surveys were collected on a representative sample of two different populations, the first (n=2103) in the province of Quebec (Canada) in 1990 (QNS) and the second in the US population (n=14 877) between 1988 and 1994 (NHANES III). MAIN OUTCOMES MEASURES: The social gradient hypothesis was tested with four different social position indicators. The analyses were performed separately for men and women aged from 18 to 74 y, for all the nutrients available in the databanks. For the USA, the graded relationship for the total population, for the non-Hispanic white population and for respondents sometimes or often experiencing a lack of food was also measured. RESULTS: A graded relationship between almost all nutrients and the studied social position measurements is observed for the consumption of total calorie-adjusted nutrients, for the proportion of people eating in accordance with dietary guidelines and for the proportion of individual not meeting 75% of the respective Recommended Dietary Allowances for their country. The direction of these relationships (positive or negative) is mainly in accordance with the clinical known impacts of nutrients on chronic diseases. CONCLUSIONS: It is possible that nutrition plays a role in the graded distribution of social health inequalities in North America even in the magnitude of this contribution remains to be evaluated. PMID- 11378811 TI - Results from two repeated 5 day dietary records with a 1 y interval among patients with colorectal polyps. AB - BACKGROUND: Dietary factors are known to be associated with initiation and development of colorectal cancer (CRC), and also with CRC's major precursor, the colorectal polyp. In long-term intervention studies on colorectal polyps, dietary changes may therefore affect potential effects of the study intervention. OBJECTIVE: To examine potential dietary changes among polyp-patients randomly selected from a 3 y intervention study after 1 y. DESIGN AND SUBJECTS: Of 116 polyp-bearing out-patients (50% men), aged 50-76 y, who participated in the double-blind 3 y placebo-controlled endoscopic follow-up and intervention study against growth and recurrence of polyps, 30 patients were randomised (strata: sex, age and polyp size) to perform a repeated 5 day dietary record by weighing after 1 y. The patients received a daily mixture of vitamin C (150 mg), alpha tocopherol (75 mg), beta-carotene (15 mg), selenium (101 microg) and calcium (1.6 g) or placebo (lactose) for a period of 3 y with annual colonoscopic examinations and polyps size measurements to test if the mixture was able to reduce polyp growth and recurrence. Polyps of >9 mm were removed, whereas the remainders and new discoveries of polyps <9 mm were left in situ until the end of the study. RESULTS: Twenty-nine patients agreed to perform the repeated 5 day dietary record, and 86% performed the second record within 48-58 weeks after the first record. The results showed that, with the exception of vitamin D, milk and milk products, no significant differences were found between the two records. The median value of the Spearman's correlation coefficient for energy and energy yielding nutrients was 0.66, for vitamins and minerals 0.58, and for foods 0.58. Individual differences between the records were found for most variables, but most of these were negligible. CONCLUSION: After 1 y, no major dietary changes were found which could be associated with a changed susceptibly for malignancy, and thereby affect potential effects of the study intervention. We may thus suggest that a potential changed susceptibility towards growth and recurrence of polyps, is due to the specific intervention, and not due to other major dietary changes. PMID- 11378812 TI - Growth status of Jordanian schoolchildren in military-funded schools. AB - OBJECTIVE: To study the growth status of Jordanian boys and girls in comparison with the Centers for Disease Control (CDC) growth charts. DESIGN: Cross-sectional study. SUBJECTS AND DATA COLLECTION: A total of 5826 boys and 1414 girls, aged 6.5-17.5 y, were included in the study. Height and weight were measured. Body mass index (BMI) was calculated as weight (kg) divided by the square of the height (m). Socioeconomic data were collected using a structured questionnaire. RESULTS: The height-for-age values fluctuated between the 5th and 10th percentiles of the CDC for both sexes, and then after the age of 8.5 and 14.5 y for boys and girls, respectively, values were just above the 10th percentile. The body weight-for-age values were just above the 25th percentile for boys and fluctuated between the 25th and 50th percentiles for girls; then after the age of 14.5 and 12.5 y for boy and girls, respectively, values fluctuated between the 25th and 50th percentiles for boys and just above the 50th percentile for girls of the CDC values. BMI values for boys were just above the 50th percentile of the CDC and for girls values fluctuated between the 50th and 75th percentiles until the age of 13.5, then values matched the 75th percentile of the CDC. CONCLUSION: The height of Jordanian children ranges from the 5th to the 10th percentile of the CDC reference values during schools years. Girls have a tendency toward obesity after puberty. PMID- 11378813 TI - Urinary iodine and thyroid status of New Zealand residents. AB - OBJECTIVES: The aim of this project was to assess the clinical significance of our low iodine excretions in terms of thyroid hormone status and thyroid volume in an adult population in a low soil iodine area of the South Island of New Zealand. DESIGN AND SETTING: Two-hundred and thirty-three residents of Otago, New Zealand collected two 24 h urine samples for assessment of iodine status. Thyroid status was determined from serum total T(4), TSH and thyroglobulin, and thyroid volumes. Relationships between urinary iodide excretion and measures of thyroid status were determined and subjects were allocated to one of three groups according to low, medium and high iodide excretion, for comparison of thyroid hormones and thyroid volumes. RESULTS: Significant correlations were found for relationships between measures of urinary iodide excretion and thyroid volume and thyroglobulin. Multiple regression analysis of data for subjects divided into three groups according to 24 h urinary iodide excretion (<60, 60-90; >90 microg iodide/day) or iodide/creatinine ratio (<40; 40-60; >60 microg/g Cr) showed significant differences in thyroid volume (P=0.029; P=0.035, respectively) and thyroglobulin (P=0.019; P=0.005, respectively) among the groups. CONCLUSIONS: The results of this study confirm the low iodide excretions of Otago residents, and indicate that the fall in iodine status is being reflected in clinical measures of thyroid status, including enlarged thyroid glands and elevated thyroglobulin. Our observations suggest the possible re-emergence of mild iodine deficiency and goitres in New Zealand. This situation is likely to worsen should iodine intakes continue to fall and continued monitoring of the situation is imperative. PMID- 11378814 TI - Body composition unaltered for African women classified as 'normal but vulnerable' by body mass index and mid-upper-arm-circumference criteria. AB - OBJECTIVE: To test the hypothesis that 'normal but vulnerable' adults, as defined by body mass index (BMI) in combination with mid-upper-arm-circumference (MUAC), are closer to normal than to malnourished ones. For that purpose body composition measurements were compared between normal and low BMI categories and according to MUAC value in an African context and for different age groups. DESIGN: Reanalysis of data from a previous cross-sectional cluster sample nutrition survey. SETTING: A rural area of the Republic of Congo, Central Africa. SUBJECTS: A representative sample (n=544) of non-pregnant women. MAIN OUTCOME MEASURES: Arm muscle area was calculated from measurements of triceps skinfold thickness and MUAC. Peripheral body fat was assessed by the sum of four skinfold thicknesses. The ratio of resistance at high and low frequencies was derived from whole body measurement of multifrequency bioelectrical impedance analysis and used as the extracellular to total body water ratio index. RESULTS: The prevalence of thinness decreased from 18.7% as defined by BMI alone to 9.0% as defined by BMI and MUAC. This difference was due to the group of subjects classified as 'normal but vulnerable' (9.7%). Prevalence of thinness increased with age when assessed by BMI alone, but no longer when assessed by BMI and MUAC. Comparison with the BMI> or =18.5 kg/m(2) category showed that in 'normal but vulnerable' subjects lower BMI was accompanied by lower both fat and lean compartments, in absolute values, but the equilibrium of body water compartments was not altered. In BMI<18.5 women, low MUAC was associated with altered lean tissues, at peripheral and whole body level, whereas fat tissue did not differ. CONCLUSIONS: 'Normal but vulnerable' subjects appeared as 'thin but healthy' rather than malnourished, at all ages, even though their BMI was lower than 18.5 kg/m(2). The new classification of thinness based on BMI and MUAC provides a more specific index of nutritional status when restricting the thin category to more at-risk subjects. PMID- 11378815 TI - Socio-economic differences in height and body mass index of children and adults living in urban areas of Karachi, Pakistan. AB - OBJECTIVE: To study the socio-economic differences in height and body mass index (BMI) in urban areas of Karachi. DESIGN: A comparative study was undertaken to compare the heights and BMIs of adults and children belonging to three distinctively different income groups living in urban areas of Karachi. SETTING: Data was collected from families living in small, medium and large houses located in the authorised urban residential areas of Karachi. SUBJECTS: A total of 600 families, 200 from each income group, were included in the study. Anthropometric measurements of 1296 females and 1197 males of different ages were taken. METHODS: All the housewives were interviewed to collect socio-demographic information. Height and weight of all the available family members were measured. In order to determine the socio-economic difference in height status, the mean height in cm of adults was compared. For children (2-17 y) means of height-for age Z-scores determined on the basis of NCHS reference values were compared. For studying the weight status the BMI of all the respondents was calculated and they were grouped into categories of under-, normal or overweight according to the NCHS recommended cut-off points. For adult men and women BMI values <18.5 kg/m(2) indicated underweight and >25 kg/m(2) indicated overweight. Among children, those having BMI values below the 5th percentile of the NHANES III reference values were categorised as underweight and those above the 95th percentile were termed overweight. RESULTS: Height status improved with income level among adults and children of both sexes. Among males the difference in weight status was significant only among 2 to 18-y-olds (P<0.05 in each case). The rate of overweight among 2 to 18-y-old males was significantly higher (P=0.004) at the middle-income level (15%) as compared to low or high income. The rate of underweight was significantly higher (P=0.025) at the low-income level among 2 to 18-y-old males (31%, 21% and 22% at low-, middle- and high-income levels, respectively). Among females, rates of underweight were not significantly different at any age. Rates of overweight increased significantly (P=0.048) with income level among 41 to 60-y-old women (38%, 53% and 60% at low-, middle- and high-income levels, respectively). CONCLUSION: Chronic undernutrition as indicated by deficit in height decreased with increasing income level. Socio economic differences in weight status were not uniform among various age-sex groups. The influence of increasing affluence is likely to be seen both in the form of increased obesity among older females and underweight among children. Differing patterns of association between income and weight status among male and female children need to studied further with more accurate birth records, so as to further clarify the situation. In terms of prevention of nutrition-related disorders both problems of under- and over-nutrition need to be addressed. PMID- 11378817 TI - A personal view on reviewing the psychological consequences of predictive genetic testing for late onset disease. PMID- 11378819 TI - A novel automated strategy for screening cryptic telomeric rearrangements in children with idiopathic mental retardation. AB - Cryptic unbalanced subtelomeric rearrangements are known to cause a significant proportion of idiopathic mental retardation in childhood. Because of the limited sensitivity of routine analyses, the cytogenetic detection of such rearrangements requires molecular techniques, namely FISH and comparative genomic hybridisation (CGH). An alternative approach consists in using genetic markers to detect segmental aneusomy. Here, we describe a new strategy based upon automated fluorescent genotyping to search for non mendelian segregation of telomeric microsatellites. A total of 29 individuals belonging to 24 unrelated families were screened and three abnormal patterns of segregation were detected (two rearrangements and one parental disomy). This study gives strong support to the view that cryptic telomeric rearrangements significantly contribute to idiopathic mental retardation and demonstrates that fluorescent genotyping is a very sensitive and cost-effective method to detect deletions, duplications and uniparental disomies. PMID- 11378820 TI - Genetic analysis in Finnish families with inflammatory bowel disease supports linkage to chromosome 3p21. AB - In inflammatory bowel diseases (IBD), certain chromosomal candidate loci have been repeatedly identified by independent studies in different populations. To investigate the contribution of the loci on chromosomes 1, 3, 7, 12, 14, and 16 to the susceptibility of IBD in Finnish population, where the predominant feature is the excess of ulcerative colitis (UC) families compared to Crohn's disease (CD) families, we carried out linkage analyses using 93 Finnish, multiply affected IBD families. We observed nominal evidence for linkage to chromosome 3p21, consistent with earlier reports. The lod scores peaked at D3S2432, with a maximum two-point lod score of 1.68 (P=0.0027). In addition, we studied whether risk of IBD is associated with functional variants of two positional candidate genes; the chemokine receptor CCR5 gene on chromosome 3p21 and the interleukin-4 receptor alpha-subunit gene (IL4RA) on chromosome 16. We did not find any significant correlation between a 32-bp deletion variant of CCR5 or a single nucleotide change A1902G (Gln576Arg) of IL4RA, and IBD phenotypes, with the exception that in the UC group homozygosity for the G1902 allele of IL4RA was less frequent (0.019 vs 0.049, P=0.038). In conclusion, our study, carried out in a genetically homogenous population, suggests that chromosome 3 may contain a susceptibility gene for IBD. PMID- 11378821 TI - Analytic power calculation for QTL linkage analysis of small pedigrees. AB - Power calculation for QTL linkage analysis can be performed via simple algebraic formulas for small pedigrees, but requires intensive computation for large pedigrees, in order to evaluate the expectation of the test statistic over all possible inheritance vectors at the test position. In this report, we show that the non-centrality parameter for an arbitrary pedigree can be approximated by the sum of the variances of the correlations between all pairs of relatives, each variance being weighted by a factor that is determined by the mean correlation of the pair. We show that this approximation is sufficiently accurate for practical purposes in small to moderately large pedigrees, and that large sibships are more efficient than other family structures under a range of genetic models. PMID- 11378822 TI - Association between quantitative traits underlying asthma and the HLA-DRB1 locus in a family-based population sample. AB - The region of human chromosome 6 containing the MHC has been identified as influencing asthma and atopy (allergy) by several genome-wide searches. The MHC contains many genes with potential effects on innate and specific immunity. As a first step in dissecting MHC influences on asthma and its underlying quantitative phenotypes, we have examined the HLA-DRB1 locus in a population sample consisting of 1004 individuals from 230 families from the rural Australian town of Busselton. The locus was strongly associated with the (log(e)) total serum IgE concentration, accounting for 4.0% of the sigma(2) (variance) in that trait (multi-allelic test, P=0.00001). The locus also influenced specific IgE titres to common allergens (multi-allelic tests, 2.8% sigma(2) for the house dust mite allergen Der p I, P=0.0013; 3.0% of sigma(2) for Der p II, P=0.0007; and 2.1% of sigma(2) for the cat allergen Fel d I, P=0.014). No associations were found to the categorical phenotype of asthma, or to the quantitative traits of peripheral blood eosinophil counts and bronchial hyper-responsiveness. Transmission disequilibrium tests excluded genetic admixture as a cause of false-positive findings. The results indicate that HLA-DRB1 alleles modulate the total serum IgE concentration and IgE responses to allergens, but do not account for the previous observations of linkage of asthma to the MHC. PMID- 11378823 TI - Genetic polymorphism of MUC7: allele frequencies and association with asthma. AB - MUC7 encodes a small salivary mucin, previously called MG2, a glycoprotein with a putative role in facilitating the clearance of oral bacteria. The central domain of this glycoprotein was previously shown to comprise five or six tandemly repeated units of 23 amino-acids which carry most of the O-linked glycans. The polymorphism of these two allelic forms (MUC7*5 or MUC7*6) has been confirmed in this study in which we have analysed a large cohort of subjects (n = 375) of various ethnic origins. We have also identified a novel rare allele with eight tandem repeats (MUC7*8). MUC7*6 was the most common allele (0.78-0.95) in all the populations tested. The tandem repeat arrays of 22 MUC7*5 alleles and 34 MUC7*6 alleles were sequenced. No sequence differences were detected in any of the MUC7*6 alleles. Twenty-one MUC7*5 alleles sequenced lacked the 4th tandem repeat (structure TR12356), while one showed the structure TR12127. The structure of the MUC7*8 allele was TR12343456. Because of the known role of MUC7 in bacterial binding, and thus its potential involvement in susceptibility to chest disease we also tested MUC7 in our previously described series of Northern European atopic individuals with and without associated asthma. The MUC7*5 allele was rarer in the atopic asthmatics than in the atopic non-asthmatics (P = 0.014, OR for no asthma in atopic individuals 3.13, CI 1.01-6.10), and the difference in frequency between all asthmatics and all non-asthmatics was statistically significant (P = 0.009) while there was no difference between atopy and non-atopy (P = 0.199). In this study we also report the electrophoretic analysis of the MUC7 glycoprotein in saliva from individuals of different MUC7 genotype. PMID- 11378824 TI - Mutational spectrum of the ED1 gene in X-linked hypohidrotic ectodermal dysplasia. AB - X-linked hypohidrotic ectodermal dysplasia (XLHED) is the most common form of the ectodermal dysplasias characterised by an abnormal development of eccrine sweat glands, hair and teeth. The ED1 gene responsible for the disorder undergoes extensive alternative splicing and to date few studies have concerned the full length transcript. We screened 52 unrelated families or sporadic cases for mutation in the full coding sequence of this gene. SSCA analysis or direct sequencing allowed identification of mutations in 34 families: one initiation defect, twenty-two missenses, two nonsense, eight insertions or deletions, and a large deletion encompassing all the ED1 gene. Fourteen of these mutations have not been previously described, including five missenses. One third of identified mutations were localised in codons 155 and 156, affecting CpG dinucleotides and nine of them correspond to the R156H missense. Hypothesis of a founder effect has been ruled out by haplotype analysis of flanking microsatellites. These recurrent mutations indicate the functional importance of the positively charged domain of the protein. Including our data, there are now 56 different mutations reported in 85 independent patients, that we have tabulated. Review of clinical features in the present series of affected males and female carriers showed no obvious correlation between the type of mutations, the phenotype and its severity. The X chromosome pattern of inactivation in leucocytes showed little correlation with expressivity of the disease in female carriers. Finally this study is useful for functional studies of the protein and to define a diagnostic strategy for mutation screening of the ED1 gene. PMID- 11378825 TI - High-resolution physical and transcript map of human chromosome 2p21 containing the sitosterolaemia locus. AB - Sitosterolaemia (phytosterolaemia) is an autosomal recessive disorder characterised by the presence of tendon xanthomas in the face of normal or mildly elevated plasma cholesterol levels, premature atherosclerotic disease and has diagnostically elevated plasma and tissue plant sterol concentrations. Affected individuals show an increased absorption of both cholesterol and sitosterol from the diet, decreased bile clearance of these sterols and their metabolites resulting in markedly expanded whole body cholesterol and sitosterol pools. The defective gene is therefore hypothesised to play a crucial role in regulating dietary cholesterol absorption, and its elucidation may shed light on these molecular processes. We have previously localised the defective gene to human chromosome 2p21, between microsatellite markers D2S1788 and D2S1352, a distance of approximately 15 cM. Recently, the disease locus interval has been narrowed to lie between D2S2294 and D2S2291/D2S2174. We have constructed a high-resolution YAC and BAC contigs by using known STSs and generating novel STSs from the minimal interval. Eight previously identified genes and 60 ESTs were mapped to these contigs. The BAC contig contains 60 BAC clones and 108 STSs and encompasses a physical distance of approximately 2.0 cM between microsatellite markers D2S2294 and D2S2291. These results will not only facilitate cloning of the sitosterolaemia gene, but also other disease genes located in this region, and accelerate sequencing of the corresponding genomic clones. PMID- 11378826 TI - Fine mapping of a gene responsible for regulating dietary cholesterol absorption; founder effects underlie cases of phytosterolaemia in multiple communities. AB - Sitosterolaemia (also known as phytosterolaemia, MIM 210250) is a rare recessive autosomal inherited disorder, characterised by the presence of tendon and tuberous xanthomas, accelerated atherosclerosis and premature coronary artery disease. The defective gene is hypothesised to play an important role in regulating dietary sterol absorption and biliary secretion, thus defining a molecular mechanism whereby this physiological process is carried out. The disease locus was localised previously to chromosome 2p21, in a 15 cM interval between microsatellite markers D2S1788 and D2S1352 (based upon 10 families, maximum lodscore 4.49). In this study, we have extended these studies to include 30 families assembled from around the world. A maximum multipoint lodscore of 11.49 was obtained for marker D2S2998. Homozygosity and haplotype sharing was identified in probands from non-consanguineous marriages from a number of families, strongly supporting the existence of a founder effect among various populations. Additionally, based upon both genealogies, as well as genotyping, two Amish/Mennonite families, that were previously thought not to be related, appear to indicate a founder effect in this population as well. Using both homozygosity mapping, as well as informative recombination events, the sitosterolaemia gene is located at a region defined by markers D2S2294 and Afm210xe9, a distance of less than 2 cM. PMID- 11378827 TI - Multiple origins of the mtDNA 7472insC mutation associated with hearing loss and neurological dysfunction. AB - Several mtDNA mutations have been reported in families with both syndromic and non-syndromic hearing loss. One such mutation is the heteroplasmic 7472insC in the tRNA(Ser(UCN)) gene which has been found in six families, all from Western Europe. However, it was not clear if this distribution was due to a common founder effect or chance sampling of several unrelated families, the 7472insC mutation having occurred multiple times. Haplotype analysis of all six families supports the latter notion. This confirms the pathogenicity of the 7472insC mutation and suggests it may exist in other populations where it may prove to be a small but significant cause of hearing loss, particularly when neurological symptoms are also present. PMID- 11378828 TI - Molecular genetic basis and prevalence of glycogen storage disease type IIIA in the Faroe Islands. AB - Glycogen storage disease type IIIA (GSD IIIA) is caused by mutations of the amyloglucosidase gene (AGL). For most populations, none of the AGL mutations described to date is particularly frequent. In this paper, we report that six children with GSD IIIA from the Faroe Islands were found to be homozygous for the novel nonsense mutation c.1222C>T (R408X) of the AGL gene. This mutation is easily detected by restriction enzyme digest with NsiI after mismatch PCR. Investigating five intragenic polymorphisms, we could show that this mutation was always associated with the same haplotype. The c.1222C>T mutation could be detected on two chromosomes of another 50 unselected GSD IIIA patients of other European or North American origin which means that this mutation plays a minor role worldwide. From the fact that we are currently aware of a total of 14 GSD IIIA cases in the Faroese population of 45 000, the observed prevalence is 1 : 3100. While the novel AGL mutation c.1222C>T was not detectable among 198 German newborns, nine out of 272 children from the Faroese neonatal screening program were found to be heterozygous for this mutation. Thus, the calculated prevalence is 1 : 3600 (95% CI 1:700-1:6400). We conclude that due to a founder effect, the Faroe Islands have the highest prevalence of GSD IIIA world-wide. The detection of the molecular defect has facilitated the diagnosis and has offered the opportunity for prenatal diagnosis in this patient group. PMID- 11378829 TI - Why are thiazide diuretics declining in popularity? PMID- 11378830 TI - Trends in antihypertensive drugs in the elderly: the decline of thiazides. AB - The last decade has seen the publication of different editions of guidelines for the pharmacological treatment of hypertension that were based on the results of large, randomised trials. Since these guidelines were meant to inform practitioners, we analysed the pattern of prescription of antihypertensive agents between 1988 and 1997 among older hospitalised adults. Because of the wealth of data supporting the use of thiazides diuretics, we focused on diuretic prescription, to identify independent predictors of their utilisation. To this end, we used the GIFA database that includes patients admitted to academic medical centres throughout Italy between 1988 and 1997. We studied 5061 patients over 65 years of age selected among a population of 28 411, based on the diagnosis of arterial hypertension at discharge. The use of ACE-inhibitors has been raising steadily through the years, and they are the agents most commonly used since 1996. Calcium channel blockers showed a similar trend and were the top prescribing drug until 1995; afterwards, the documentation of potentially severe side effects has resulted in a nearly 20% reduction of their use. Beta-blockers have remained unpopular throughout the decade. Instead, the prescription of diuretics as a class showed a biphasic trend; an initial decrease with a prolonged steady state and a more recent raise. However, at a separate analysis, it was a evident that a progressive increase of the use of loop diuretics since 1988 has been paralleled by a nearly 50% reduction of thiazides prescriptions. Loop diuretics were more likely to be prescribed to older individuals, those with cardiac heart failure, coronary heart disease and high creatinine level. In contrast, independent predictors of thiazides use were female gender, good functional status, preserved renal function, and absence of cardiovascular comorbidity. In conclusion, despite continued recommendations to use thiazides diuretics for the treatment of hypertension among older individuals, their use has been declining steadily between 1988 and 1997. A possible explanation is that the choice to prescribe a thiazides diuretic is influenced by age, functional status and comorbidity. PMID- 11378831 TI - Apolipoprotein B, ratio of total cholesterol to HDL-C, and blood pressure in abdominally obese white and black American women. AB - OBJECTIVE: To compare the association of apolipoprotein B (ApoB) and total cholesterol to high-density lipoprotein cholesterol (TC/HDL) with blood pressure in abdominally obese white and black American women. We also sought to determine if there are ethnic differences in blood pressure values that could be explained by differences in mean values of ApoB and TC/HDL. METHODS: Data (n = 1844) from the Third US National Health and Nutrition Examination Survey were used in this study. Abdominal obesity was defined as waist circumference (WC) of > or =88cm or having WC greater than what was expected as predicted from residuals obtained from linear regression of WC on BMI. Bi-variate Pearson's correlation analysis was used to quantify the degree of association of ApoB and TC/HDL with blood pressure and other lipids. Multiple linear regression analysis was used to assess the independent contribution of ApoB and TC/HDL to blood pressure, adjusting for age, total cholesterol, alcohol intake, and smoking. To determine ethnic differences in blood pressure values associated with ApoB or TC/HDL, dummy variables were used to compare blacks with whites fitted in multiple regression models, while adjusting for age, total cholesterol, alcohol intake and smoking. RESULTS: Elevated ApoB was positively associated with diastolic and systolic blood pressure (DBP/SBP) in blacks and whites, independent of age, total cholesterol, alcohol intake and smoking (P < 0.01). Elevated TC/HDL was also positively associated with increased DBP and SBP in whites (P < 0.05). For the same value of ApoB and TC/HDL whites had higher values of DBP and SBP than blacks, adjusting for confounding variables. CONCLUSIONS: Compared with TC/HDL, ApoB was more strongly associated with DBP and SBP in both abdominally obese white and black women. Since ApoB is associated with hypertension, the combination of elevated ApoB and hypertension may identify a group of patients with more marked risk of vascular disease, thus, warranting further investigation. PMID- 11378832 TI - Abdominal obesity defined as a larger than expected waist girth is associated with racial/ethnic differences in risk of hypertension. AB - OBJECTIVE: Waist circumference (WC) cut-points of > or =102 cm and > or =88 cm for men and women, respectively, representing abdominal obesity have been recommended for determining obesity related co-morbidities. However, these cut points carry the component of generalised obesity estimated by body mass index (BMI). The aim of this investigation was to determine whether abdominal obesity free of the influence of overall heaviness is associated with increased risk of hypertension in a representative sample of white and black Americans. METHODS: Data (n = 11114) from the Third US National Health and Nutrition Examination Survey were used in this investigation. Standardised residual values from the linear regression of WC on BMI were used to define abdominal obesity status. The risk of hypertension associated with abdominal obesity was estimated from the logistic regression model, adjusting for age, smoking and alcohol. We also estimated the public health consequences of abdominal obesity from the population attributable fraction of hypertension. RESULTS: Relative to white, black race/ethnicity was associated with approximately 1.8 and approximately 2.7 greater risk of hypertension in men and women, respectively, adjusting for abdominal obesity, age, smoking and alcohol consumption. Having larger than expected waist girths were associated with 1.58 and 1.39 increased risk of hypertension in black men and black women, respectively, adjusting for confounders. Population attributable risks of hypertension due to abdominal obesity were approximately 24.9% and 15.9%, in black men and black women, respectively. CONCLUSIONS: In Americans, hypertension is a public health problem that is closely linked to abdominal adiposity. An important research challenge therefore is to determine the best way to regulate body weight under conditions of food abundance. There is a need to clarify how lifestyle habits promote large waist sizes leading to abdominal adiposity and associated cardiovascular disease in the US, particularly among black Americans. PMID- 11378833 TI - Dose response effect of cyclical medroxyprogesterone on blood pressure in postmenopausal women. AB - OBJECTIVE: This study was designed to compare with placebo the dose-response effect of cyclical doses of the C21 progestogen, medroxyprogesterone acetate (MPA) on blood pressure (BP) when administered to normotensive postmenopausal women receiving a fixed mid-range daily dose of conjugated equine oestrogen (CEE). MATERIALS AND METHODS: Twenty normotensive postmenopausal women (median age 53 years) participated in the study which used a double-blind crossover design. There were four randomised treatment phases, each of 4 weeks duration. The four blinded treatments were MPA 2.5 mg, MPA 5 mg, MPA 10 mg and matching placebo, taken for the last 14 days of each 28 day treatment cycle. CEE 0.625 mg was also administered once daily as open labelled tablets to all subjects throughout the study. Clinic BP was measured weekly with the mean values of weeks 3 and 4 of each phase used for analysis. Ambulatory BP was performed in the final week of each phase. RESULTS: Compared with the placebo phase, end of phase clinic BP was unchanged by any of the progestogen treatments. There was a dose-dependent decrease in ambulatory daytime diastolic and mean arterial BP with the progestogen treatments compared with placebo (P < 0.05). CONCLUSION: In a regimen of postmenopausal hormone replacement therapy with a fixed mid-range daily dose of CEE combined with a cyclical regimen of a C21 progestogen spanning the current clinical dose range, the progestogen has either no effect or a small dose dependent reduction in clinic and ambulatory BPs over one treatment cycle. PMID- 11378834 TI - Circadian rhythm and postural change in natriuresis in non-dipper type of essential hypertension. AB - Recently we found that, in non-dipper type of essential hypertensive patients who showed the lack of nocturnal fall in blood pressure, circadian rhythm of urinary sodium excretion rate was disturbed. In the present study we examined whether postural change in natriuresis is also disturbed in non-dippers. Sixteen inpatients with essential hypertension were maintained on a relatively high sodium diet containing 10 to 12 g of NaCl per day for 8 days. On the 7th day of the study period, 24-h ambulatory blood pressures were measured, and 5-7th days urinary samples were collected for both daytime and night-time. On the last day of the study period, patients stood for 2 h and then lay down for 2 h. Urinary volume and excretion rates of creatinine and sodium were measured every hour in both positions. Night-time urinary sodium excretion rate was significantly higher in non-dippers (n = 9) than that in dippers (n = 7). Night/day ratio of mean arterial pressure had a positive relationship with night/day ratio of urinary sodium excretion rate. In non-dippers, but not in dippers, the mean value of U(Na)V during upright position was significantly lower than that during supine position. There was a significantly negative relationship between upright/supine ratio of U(Na)V and night/day ratio of MAP or night/day ratio of U(Na)V. In patients with non-dipper type of essential hypertension, both natriuresis patterns, circadian rhythm and postural change, were disturbed. PMID- 11378835 TI - The PANDORA project: results of the cost of illness analysis. AB - AIMS: To evaluate the cost of illness from hypertension for the Italian National Health System (NHS). METHODS AND RESULTS: A prospective analysis was carried out on clinical and economic data recorded in the general practitioners' (GPs) database. Twenty-one GPs working in the Ravenna area in Italy took part in the project on a voluntary basis. The study included 1047 hypertensive patients enrolled between 1 June and 31 December 1997 and continued for 365 days from the date of enrolment. The following costs were calculated: antihypertensive drugs, laboratory tests and instrumental procedures, GP visits for blood pressure control, specialist visits, casualty visits, hospitalisation due to cardiovascular problems. In the whole sample, the most relevant cost is due to antihypertensive drugs (42.7%), followed by hospital admission (28.4%), GP visits (15.1%) and tests (10.6%). The total mean cost was significantly lower in incident (no previous treatment) than in prevalent patients (already treated) (457 512 vs 725 573 Italian Lira (ITL), P < 0.05) and in older rather than in younger patients (1171 410 vs 796 452 (ITL) P < 0.05). (In the text the equivalent is given in Euros, Pounds Sterling and US dollars). CONCLUSION: Our study should be considered as preliminary, nevertheless it could represent a step towards the evaluation of the true cost of hypertension. PMID- 11378836 TI - Association study of the 5-HT(2A) receptor gene polymorphism, T102C and essential hypertension. AB - BACKGROUND: Serotonin dysfunction has been implicated in hypertension due to its ability to induce vasoconstriction via stimulation of 5-HT(2) receptors and due to the antihypertensive effect of ketanserin, an antagonist at the 5-HT(2A) receptor subtype, expressed both on arteries and the brain. The silent T102C polymorphism in the 5-HT(2A) gene is in absolute linkage disequilibrium with a polymorphism in the promoter and may contribute to genetic predisposition possibly by modifying the transcription of the gene. OBJECTIVE: To examine the genetic contribution of the T102C 5-HT(2A)polymorphism in essential hypertension in a case-control sample of UK residents. DESIGN: The hypertensive group consisted of 342 subjects over 75 years and the community-based control group consisted of 319 subjects. Subjects were genotyped for the T102C polymorphism by Mspl restriction enzyme digestion following PCR amplification. RESULTS: Sex specific association analysis revealed significant differences between hypertensive and normotensive subjects in the genotypes distribution (P = 0.016) and allelic frequencies (P = 0.007) in the female group. The direction of significance was increased frequency of the 102-C allele in the hypertensive subjects. There were no association between haplotype and age or body mass index, which suggest that the effect of the T102C variant is not influenced by these variables. CONCLUSION: This data indicates that the T102C polymorphism in the 5 HT(2A) gene might be an independent risk factor for increased blood pressure in female individuals with essential hypertension. PMID- 11378837 TI - Genetic bottlenecks, perceived racism, and hypertension risk among African Americans and first-generation African immigrants. AB - The complexity of factors influencing the development of hypertension (HTN) in African Americans has given rise to theories suggesting that genetic changes occurred due to selection pressures/genetic bottleneck effects (ie, constriction of existing genetic variability) over the course of the slave trade. Ninety-nine US-born and 86 African-born health professionals were compared in a cross sectional survey examining genetic and psychosocial predictors of HTN. We examined the distributions of three genetic loci (G-protein, AGT-235, and ACE I/D) that have been associated with increased HTN risk. There were no significant differences between US-born African Americans and African-born immigrants in the studied genetic loci or biological variables (eg, plasma renin and angiotensin converting enzyme activity), except that the AGT-235 homozygous T genotype was somewhat more frequent among African-born participants than US-born African Americans. Only age, body mass index, and birthplace consistently demonstrated associations with HTN status. Thus, there was no evidence of a genetic bottleneck in the loci studied, ie, that US-born African Americans have different genotype distributions that increase their risk for HTN. In fact, some of the genotypic distributions evidenced lower frequencies of HTN-related alleles among US-born African Americans, providing evidence of European admixture. The consistent finding that birthplace (ie, US vs Africa) was associated with HTN, even though it was not always significant, suggests potential and unmeasured cultural, lifestyle, and environmental differences between African immigrants and US-born African Americans that are protective against HTN. PMID- 11378838 TI - The significance of hypertensive response to exercise as a predictor of hypertension and cardiovascular disease. AB - Several studies have shown that exaggerated blood pressure (BP) response to exercise can predict the development of hypertension and target organ damage, but others did not. The aim of this study was to evaluate the relationship between exaggerated BP response to exercise (ExBPR) and the development of hypertension or cardiovascular disease. We reviewed the charts of male subjects who, during the years 1991-1994, had a routine check up that included an exercise stress test. For each subject, the following parameters were noted: age; body mass index (BMI); history of diabetes; cigarette smoking; family history of ischaemic heart disease and lipid profile. BP and heart rate at rest and during exercise were recorded as well. ExBPR was defined whenever peak exercise systolic BP or diastolic BP was over 200 mm Hg or 100 mm Hg respectively. We identified 73 males who exhibited ExBPR and matched them with a control group of 117 subjects with similar age who had a normal BP response. The mean age of the studied group was 42.6 years and the average follow-up was 5.7 years. Baseline characteristics were similar in both groups. During the follow-up we observed hypertension among 22% in those with ExBPR in contrast to 2.6% in the control group (P < 0.0001). In addition, more subjects of the ExBPR group required cardiovascular medications than of those in the control group (19.2% vs 4.3%, P = 0.0008). Thus, it seems that ExBPR predicts the development of hypertension and cardiovascular diseases. We therefore suggest that subjects with ExBPR should be followed more closely and be instructed for lifestyle modifications which may delay the development of such diseases. PMID- 11378840 TI - Effects of terminal digit preference on the proportion of treated hypertensive patients achieving target blood pressures. PMID- 11378841 TI - Allelic variation of a BalI polymorphism in the DRD3 gene does not influence susceptibility to bipolar disorder: results of analysis and meta-analysis. AB - Bipolar disorder is a major psychiatric illness that has evidence for a significant genetic contribution toward its development. In recent years, the BalI RFLP (restriction fragment length polymorphism) in the dopamine D3 receptor gene has been examined as a possible susceptibility factor for both schizophrenia and bipolar disorder. While analysis in schizophrenia has produced examples of increased homozygosity in patients, less encouraging results have been found for bipolar disorder. Recently, however, a family-based association study has found a significant excess of allele 1 and allele 1-containing genotypes in transmitted alleles to bipolar probands over nontransmitted controls. In a large bipolar case control sample (n = 454), we have been unable to replicate the family-based association study (chi-square = 0.137, P = 0.71, 1 df) or detect an effect similar to the positive homozygosity findings in schizophrenia (chi-square = 0.463, P = 0.50, 1 df). A meta-analysis of previous association studies also revealed no difference in allele distributions between bipolar patients and controls for this polymorphism in ethnically homogeneous samples (odds ratio, OR, = 1.04; P = 0.60; 95% confidence interval, CI, = 0.89-1.20). In view of this evidence, we conclude that variation at the BalI RFLP is not an important factor influencing the susceptibility to bipolar disorder. It remains possible, however, that other sequence variations within the DRD3 gene could play a role. PMID- 11378842 TI - Phenylethanolamine N-methyltransferase (PNMT) gene and early-onset Alzheimer disease. AB - The activity of human phenylethanolamine N-methyltransferase (PNMT) is reduced in the neurons of those cells in many subcortical areas of the brain that are known to undergo neurodegeneration in Alzheimer disease (AD). Others have reported that PNMT is decreased in brains of persons with AD and that the decrease in enzymatic activity is due to a reduced amount of the enzyme protein. We have previously described two polymorphisms, G-353A and G-148A, in the promoter region of the gene coding for PNMT. These markers were tested for their association with the occurrence of sporadic AD. Genotyping of 131 necropsy confirmed AD cases, and 947 adult nondemented controls were completed. We observed a significant association between both of the PNMT gene polymorphisms and early-onset AD (EOAD) (P < or = 0.007), but not in late-onset AD (LOAD). These data suggest that genetic variation in the promoter of the PNMT gene is associated with increased susceptibility to the sporadic form of EOAD. PMID- 11378843 TI - GABA-A receptor beta3 and alpha5 subunit gene cluster on chromosome 15q11-q13 and bipolar disorder: a genetic association study. AB - There is accumulated evidence that the genes coding for the receptor of gamma aminobutyric acid (GABA), the most important inhibitory neurotransmitter in the CNS, may be involved in the pathogenesis of affective disorders. In a previous study, we have found a genetic association between the GABA-A receptor alpha5 subunit gene locus (GABRA5) on chromosome 15q11-of 13 and bipolar affective disorder. The aim of the present study was to examine the same subjects to see if there exists a genetic association between bipolar affective disorder and the GABA receptor beta3 subunit gene (GABRB3), which is located within 100 kb from GABRA5. The sample consisted of 48 bipolar patients compared to 44 controls (blood donors). All subjects were Greek, unrelated, and personally interviewed. Diagnosis was based on DSM-IV and ICD-10 criteria. The marker used was a dinucleotide (CA) repeat polymorphism with 12 alleles 179 to 201 bp long; genotyping was successful in all patients and 43 controls. The distribution of GABRB3 genotypes among the controls did not deviate significantly from the Hardy Weinberg equilibrium. No differences in allelic frequencies between bipolar patients and controls were found for GABRB3, while this locus and GABRA5 did not seem to be in significant linkage disequilibrium. In conclusion, the GABRB3 CA repeat polymorphism we investigated does not present the observed association between bipolar affective illness and GABRA5. This could be due to higher mutation rate in the GABRB3 CA-repeat polymorphism, but it might also signify that GABRA5 is the gene actually associated with the disease. PMID- 11378844 TI - Affected sibling pair linkage analysis of qualitative and quantitative traits for schizophrenia on chromosome 22 in a Chinese population. AB - We performed nonparametric linkage analysis on 136 families with two or more siblings with schizophrenia from Sichuan, southwestern China. In addition to categorical diagnosis, we used quantitative trait information from the Positive and Negative Symptom Scale and the modified Overt Aggression Scale. Categorical analysis using the diagnosis of schizophrenia and a maximum likelihood identity by-descent method produced scores of close to 0 throughout the whole region tested. Multipoint analysis allowed exclusion of most markers with a relative risk of > 2, but did not exclude the possibility of a relative risk of < 1.5 for four of the markers. Our results provide no significant evidence for a locus for schizophrenia on chromosome 22. Quantitative linkage analysis using the PANSS-G scale score produced a maximum LOD score of approximately 1.2 with the marker D22S310, using either the Haseman-Elston method or maximum likelihood variance estimation with or without dominance. PANSS-N produced a maximum LOD score of 1.2 at the D22S283 locus. LOD score of about 1 are easily produced by chance. Thus, we conclude that under quantitative trait we also find no evidence of linkage between schizophrenia and markers on chromosome 22 in our Chinese sibling pair sample. PMID- 11378845 TI - No evidence for linkage by transmission disequilibrium test analysis of microsatellite marker D22S278 and schizophrenia in a Palestinian Arab and in a German population. AB - Linkage for a schizophrenia susceptibility locus on chromosome region 22q12-q13 was initially suggested by independent studies from two groups and confirmed in a combined analysis of data for the microsatellite marker D22S278 in multiply affected schizophrenic families derived from 11 independent research groups worldwide. In addition to these reports of linkage to schizophrenia on chromosome 22, bipolar disorder has also been linked to markers in this chromosomal region. We now report results from an analysis of 223 Palestinian Arab trios from three different centers in Israel and Palestine using the allele-wise extended transmission disequilibrium test for multiallelic markers. No evidence for linkage is observed in the entire group or in any of the three centers (entire group: chi-square = 5.59, P = 0.78, df = 9; Afula: chi-square = 6.51, P = 0.48, df = 7; Bethlehem: chi-square = 14.11, P = 0.12, df = 9; Beersheva: chi-square = 7.04, P = 0.32, df = 6). Additionally, we examined D22S278 in a group of 114 schizophrenic German triads and failed to observe evidence for linkage (chi square = 8.13, P = 0.42, df = 8df). PMID- 11378847 TI - No evidence of an association between 5HT1B receptor gene polymorphism and suicide victims in a Japanese population. AB - Serotonergic systems have been reported to mediate the control of aggression and/or impulsivity in humans and to be involved in suicidal behavior. Neurochemical studies showing serotonergic dysfunction in suicide appear to support the functional alteration of serotonergic systems due to gene polymorphisms. Knock-out mice of the 5HT1B receptor gene have been reported to result in increased aggression. We hypothesized that the 5HT1B receptor-mediated serotonergic dysfunction was implicated in suicide through disinhibition of aggression and/or impulsivity. To explore this hypothesis, we examined the association between suicide victims who completed suicide and the 5HT1B receptor gene G861C polymorphism. No significant differences in genotype distribution and allele frequencies were found between suicide victims and controls. Though there is the possibility of failing to detect small effects, these results show no evidence of an association between the 5HT1B receptor gene G861C polymorphism and suicide victims in a Japanese population and indicate that it is unlikely that the 5HT1B receptor is implicated in the susceptibility to suicide. PMID- 11378846 TI - Investigation of association of 13 polymorphisms in eight genes in southeastern African American Alzheimer disease patients as compared to age-matched controls. AB - Alzheimer disease (AD) is an emotionally devastating and exceptionally costly disease. Apolipoprotein E (APOE) is a major risk factor gene for AD regardless of age of onset or family history. However, this association may not be as strong or consistent in ethnic groups such as African Americans, raising the possibility of other modifier gene(s). In a group of African American AD patients, a significantly increased risk of AD was associated with two E4 alleles (OR = 5.6; 95% CI = 1.5-21.0) or one E4 allele (OR = 2.5; 95% CI = 1.3-5.0) when compared to E3/E3 genotype, and there was a significant lowering of age of onset for affecteds with E4/E4 genotype as compared to one E2 allele (P = 0.02) or all others (P = 0.03). We also found a significant increase in age of onset with the 308 #2 (A) allele of TNF when compared to AD cases with no #2 allele. A significant increase in age was also demonstrated with the #2 allele (99 base pairs) of the microsatellite TNFa, located approximately 10.5 kb upstream of TNF. When these two alleles were combined with the TNF -238G (#1) allele to give a haplotype, the significant increase in age was still demonstrated. Polymorphisms in the APOE promoter and six other candidate genes did not appear to demonstrate any significant association with our African American AD patients. Our results confirm the established association of APOE4 to AD observed in several ethnic groups, including African Americans. In addition, TNF appears to have some modifying effect in AD, primarily on age of onset, or it could be in linkage disequilibrium with a modifier locus nearby. PMID- 11378848 TI - Association of a polymorphism of the dopamine transporter gene with externalizing behavior problems and associated temperament traits: a longitudinal study from infancy to the mid-teens. AB - There have been reports that a variable number of tandem repeats (VNTR) polymorphism situated in the 3' untranslated region of the dopamine transporter gene is associated with attention-deficit hyperactivity disorder. On the basis of these findings, we predicted an association of this polymorphism with hyperactivity, other externalizing behavior problems, and related temperament traits in a general population sample. The association was investigated using children participating in a longitudinal study of childhood temperament and development. DNA was taken from 660 children who had been assessed for temperament from 4-8 months to 15-16 years, and for behavior problems from 3-4 to 15-16 years. No significant associations were found at any age. There are a number of methodological differences from earlier studies that might explain the lack of associations with hyperactivity. It is also possible that the earlier findings are not replicable. PMID- 11378849 TI - Caught in the trio trap? Potential selection bias inherent to association studies using parent-offspring trios. AB - During the last years, the validity of classic case control studies in psychiatric genetic research has been increasingly under question due to the risk of population stratification problems inherent to this type of association study. By consequence, the application of family-based association studies using parent offspring trios has been strongly advocated. Recently, however, in a study comparing clinical characteristics between index patients from parent-offspring trios and singleton patients with bipolar affective disorder, the question was raised whether a systematic neglect of case control association studies could lead to a selection bias of susceptibility genes. In a similar approach, we compared demographic and clinical characteristics of 122 singleton bipolar patients with those of 54 bipolar patients derived from parent-offspring trios. The singleton patients did not only present with a higher age of onset, but also with a higher frequency of suicidal behavior and a higher familial loading for suicidality. These findings suggest that the genetic mechanism for disease might be different between trio-based and classic case control samples, where patients are examined whose parents are not available for genetic studies. Thus, giving up case control designs for the sake of family-based association studies could be at the risk of selecting against several genetically determined factors. PMID- 11378850 TI - Velo-cardio-facial syndrome: Implications of microdeletion 22q11 for schizophrenia and mood disorders. AB - Velo-cardio-facial syndrome (VCFS) is a congenital malformation syndrome with variable phenotypic features that has been associated with chromosomal microdeletion 22q11.2. Psychiatric disorders have been reported to be highly prevalent in individuals with this syndrome, and the objective of this study was to assess the nature and extent of psychopathology among individuals with VCFS. We studied 20 children and adolescents with 22q11 deletions determined by fluorescence in situ hybridization (FISH). Control subjects were 11 nondeleted siblings who were the closest age match to the affected subjects. Both affected and control subjects were assessed using two standardized psychiatric research instruments. The results of this study confirmed the high rate of psychiatric disorders among VCFS subjects (60% of our subjects). Of the specific types of disorders, only mood disorders were significantly more common among VCFS subjects compared to sibling controls, with eight VCFS subjects having mood disorders compared with none of the control subjects (P<0.02). Three affected subjects had schizotypal traits comorbid with a mood disorder. In addition, disruptive behavior disorders were frequently diagnosed among VCFS subjects. Using a dimensional measure of psychopathology, significant differences between VCFS subjects and sibling controls were found on three scales: ADHD (P<0.02), separation anxiety (P<0.02), and depression (P<0.01). VCFS subjects were achieving significantly less well academically and requiring significantly more special educational assistance than sibling controls. Follow-up data were available on two subjects, both of whom had been diagnosed with schizophrenia. Further research on psychopathology in VCFS may provide a model of how a specific genetic defect can lead to the development of psychiatric disorders. PMID- 11378851 TI - Serotonin transporter promoter polymorphism is associated with attenuated prolactin response to fenfluramine. AB - Disturbances in central serotonin (5-HT) function may have a role in impulsive aggression in patients with a wide range of psychiatric diagnoses. The underlying mechanism, however, remains unknown. There are several naturally occurring mutations in the 5-HT signaling pathway that may underlie differences in 5-HT function and responsivity to drugs that affect 5-HT functioning. In the present study, we examined the relationship between polymorphisms in the promoter region of the gene coding for the neuronal 5-HT transporter, fenfluramine-induced prolactin release, and aggressive impulsivity (as measured by Barratt Impulsivity Scale, Buss-Durkee Hostility Inventory, and Brown-Goodwin Aggression Scale scores), in a group of abstinent alcoholic patients and healthy volunteers. We report here that possession of the short variant of the 5-HT transporter promoter polymorphism was associated with a blunting of overall central 5-HT function, as measured by fenfluramine-induced prolactin release. We found no relationship between aggressive, hostile, or impulsive traits and fenfluramine-induced prolactin release or between these traits and polymorphisms in the 5-HT transporter promoter. Thus, we have shown that a 5-HT transporter promoter genotype, which has previously been associated with anxiety-based behaviors, alters an in vivo measure of central 5-HT function (fenfluramine-induced prolactin release), providing an important mechanism for linkage between a gene, physiological function, and behavior. Published 2001 Wiley-Liss, Inc. PMID- 11378852 TI - Scanning of estrogen receptor alpha (ERalpha) and thyroid hormone receptor alpha (TRalpha) genes in patients with psychiatric diseases: four missense mutations identified in ERalpha gene. AB - Estrogen and thyroid hormones exert effects on growth, development, and differentiation of the nervous system. Hormone administration can lead to changes in behavior, suggesting that genetic variants of the estrogen receptor alpha (ERalpha) and the thyroid hormone receptor alpha (TRalpha) genes may predispose to psychiatric diseases. To investigate this possibility, regions of likely functional significance (all coding exons and flanking splice junctions) of the ERalpha and TRalpha genes were scanned in patients with schizophrenia (113), along with pilot studies in patients with bipolar illness (BPI), puerperal psychosis, autism, attention-deficit hyperactivity disorder (ADHD), and alcoholism. A total of 1.18 megabases of the ERalpha gene and 1.16 megabases of the TRalpha gene were scanned with Detection of Virtually All Mutations-SSCP (DOVAM-S), a method that detects virtually all mutations. Four missense mutations, seven silent mutations and one deletion were identified in the ERalpha gene, while only four silent mutations were present in the TRalpha gene. Two of the missense mutations in ERalpha are conserved in the six available mammalian and bird species (H6Y, K299R) and a third sequence variant (P146Q) is conserved in mammals, birds, and Xenopus laevis, hinting that these sequence changes will be of functional significance. These changes were found in one patient each with BPI, puerperal psychosis, and alcoholism, respectively. Analysis of the ERalpha and TRalpha genes in 240 subjects reveals that missense changes and splice site variants are uncommon (1.7% and 0%, respectively). Further analyses are necessary to determine if the missense mutations identified in this study are associated with predisposition or outcome for either psychiatric or nonpsychiatric diseases. PMID- 11378853 TI - Fine mapping supports previous linkage evidence for a bipolar disorder susceptibility locus on 13q32. AB - A region between D13S71 and D13S274 on 13q32 showed linkage to bipolar disorder (BP) based on a genome scan using markers with an average spacing of approximately 6 cM and an average heterozygosity of approximately 60% [Detera Wadleigh et al., 1999: Proc Natl Acad Sci USA 96:5604-5609]. In an attempt to confirm this finding and achieve fine mapping of the susceptibility region, nine additional microsatellite markers with average heterozygosity of approximately 86%, located between D13S71 and D13S274, were typed in the same sample. The strongest linkage evidence was detected by multipoint linkage analysis (ASPEX program) around D13S779-D13S225 with maximum LOD score of 3.25 under Affection Status Model II (ASM II; P = 0.0000546). Data from additional nine markers resulted in a decrease of the 95% confidence interval of the linkage region. Association analyses with GASSOC TDT and ASPEX/sib_tdt detect potential linkage disequilibrium with several markers, including D13S280 (ASPEX TDT P = 0.0033, ASM I). These data generated using a higher marker density within the proposed susceptibility region strengthen the validity of our previous findings and suggest a finer localization of the susceptibility gene(s) on 13q32. PMID- 11378854 TI - Evidence for an association with the serotonin transporter promoter region polymorphism and autism. AB - We have examined three functional polymorphisms, serotonin transporter promoter region polymorphism (5-HTTLPR), dopamine D4 exon III repeat region (DRD4), and catechol-O-methyltransferase (COMT), in a small family-based design toward identifying candidate genes that confer risk for autism. A significant excess of the long/long 5-HTTLPR genotype was observed (likelihood ratio = 7.18; P = 0.027; 2 df; n = 33 families) as well as preferential transmission of the long allele of the 5-HTTLPR (TDT chi-square = 5.44; P<0.025; 1 df). No association was observed between the COMT and DRD4 polymorphisms and autism in this sample. Some previous studies have observed linkage between autism and the 5-HTTLPR polymorphism and the current results are similar to those first reported by Klauck et al. [1997: Hum Genet 100:224-229; 1997: Hum Mol Genet 6:2233-2238]. Additionally, elevated serotonin levels have been consistently found in 30%-50% of autistic patients and may represent a marker for familial autism. Hyperserotonemia in autism appears to be due to enhanced 5-HT uptake, as free 5-HT levels are normal and the current report of an excess of the long/long 5-HTTLPR genotype in autism could provide a partial molecular explanation for high platelet serotonin content in autism. PMID- 11378855 TI - QTL association analysis of the DRD4 exon 3 VNTR polymorphism in a population sample of children screened with a parent rating scale for ADHD symptoms. AB - Current developments in molecular genetics have led to a rapid increase in research aimed at the identification of genetic variation that influences complex human phenotypes. One phenotype that has aroused a great deal of interest is the behavioral trait hyperactivity and the related clinical disorder attention deficit hyperactivity disorder (ADHD). The driving force behind the molecular genetic research in this area is the overwhelming evidence from quantitative genetic studies that show high heritablility (h(2) = 0.7-0.9) for the behaviors characterizing the diagnosis of ADHD, whether the disorder is viewed as a categorical entity or a continuous trait. To date, molecular studies have aimed at identifying susceptibility genes for ADHD, defined using operational diagnostic criteria, and have focused on variation within genes that regulate dopamine neurotransmission. Several studies report ADHD to be associated with the 7-repeat allele of a 48 bp repeat polymorphism (DRD4-7) in exon 3 of the dopamine D4 receptor gene (DRD4). In this study, we take a dimensional perspective of ADHD and examine the relationship of this DRD4 polymorphism in a sample of children selected from the general population on the basis of high and low scores on the five ADHD items of the Strengths and Difficulties Questionnaire (SDQ) as rated by their parents. We found a significant relationship between DRD4-7 and high scoring individuals [chi-square = 8.63; P = 0.003; OR = 2.09 (95% CI 1.24 < OR < 3.54), F-statistic = 7.245; P = 0.008]. PMID- 11378856 TI - No evidence of linkage or association between ADHD and DXS7 locus in Irish population. PMID- 11378857 TI - Aggressive behavior and Brunner syndrome: no evidence for the C936T mutation in a population sample. PMID- 11378858 TI - Flow cytometric assessment of transduction efficiency and cytotoxicity of herpes simplex virus type 1-based amplicon vectors. AB - BACKGROUND: In this study, we compared herpes simplex virus type 1 (HSV-1) amplicon vector stocks prepared by transient cotransfection with two different BAC-cloned packaging-defective HSV-1 helper genomes, fHSVDeltapacDelta27 and fHSVDeltapac, with respect to transduction efficiency and cytotoxicity. Both fHSVDeltapacDelta27 and fHSVDeltapac are packaging defective because the pac signals have been deleted; fHSVDeltapacDelta27 contains an additional deletion in the HSV-1 ICP27 gene, which increases the safety of the system. METHODS: HSV-1 amplicon pHSVGFP under the control of the HSV-1 immediate-early (IE) 4/5 promotor was packaged into virus particles by transient cotransfection with either fHSVDeltapacDelta27 or fHSVDeltapac DNA. Cultures were infected with the two different vector stocks and examined under the fluorescence microscope and analyzed by flow cytometry over a 5-day period to assess transduction efficiency and cytotoxicity. RESULTS: Both vector stocks, pHSVGFP[fHSVDeltapacDelta27] and pHSVGFP[fHSVDeltapac], efficiently transduced the target cells. Interestingly, the highest mean fluorescence intensities were measured at 1 day after infection, whereas the number of GFP-fluorescent cells reached a peak at day 3 after infection. At day 3 after infection, a slight increase in the number of dead cells was observed in those cultures transduced with high doses of vector stock. Between days 3 and 4 after infection, the number of dead cells increased dramatically in all the cultures, transduced and nontransduced. Only the cultures infected with a high dose of pHSVGFP[fHSVDeltapac] displayed a significant further increase in the number of dead cells between days 4 and 5 postinfection. CONCLUSIONS: Flow cytometry allowed comparison of transduction efficiency and cytotoxicity mediated by the two different amplicon vector stocks. Cultures infected with pHSVGFP[fHSVDeltapacDelta27] were more viable than those infected with pHSVGFP[fHSVDeltapac](P < 0.05). The practical implications of this study are at the level of vector design. Flow cytometry has proven a fast and reliable approach to assess the quality of potential gene transfer vectors prior to their use in (pre) clinical trials. PMID- 11378859 TI - A flow cytometry technique for measuring chromosome-mediated gene transfer. AB - BACKGROUND: Using artificial chromosome expression systems (ACes), we have developed a unique and rapid screening technique to quantify delivery of foreign DNA into cells in vitro. Delivery was measured within 24 h after transfection, using flow cytometry to detect the transfer of ACes labeled with thymidine analogue. This technique can be used to optimize delivery parameters of ACes and heterologous DNA into cells and eventually tissue. METHOD: Chinese hamster ovary (CHO) cells carrying artificial chromosomes were grown in media supplemented with iododeoxyuridine (IdUrd). The 60-mb artificial chromosome was purified by flow cytometry sorting and transfected into Chinese hamster lung fibroblast cells (V79 4) or mouse connective tissue cells [LM(tk-)] using LipofectAMINE 2000trade mark, a cationic lipid, and Superfecttrade mark, a cationic dendrimer. The cells were incubated with an FITC-conjugated anti-bromodeoxyuridine (BrdUrd) antibody and analyzed by flow cytometry. IdUrd-incorporated artificial chromosome expressing green fluorescent protein (GFP) was transfected into V79-4 cells. Delivery was measured at 24 h and GFP expression was detected at 48 h. RESULTS: The delivery of intact artificial chromosomes into V79-4 and LMtk- cells was detected within 2 h and up to 48 h post-transfection. Maximum delivery rates of 20% and 14% were observed using LipofectAMINE 2000 and Superfect, respectively. Flow cytometry data correlated with microscopic observations. IdUrd incorporation resulted in less quenching after staining with Hoechst 33258 and chromomycin A3 than BrdUrd incorporation. The fluorescence intensity of the FITC-conjugated anti-BrdUrd antibody was greater with IdUrd-incorporated chromosomes than with BrdUrd incorporated chromosomes. CONCLUSION: The results indicate that IdUrd-labeled artificial chromosomes can be detected 24 h after transfection. This efficient, sensitive, high-throughput detection technique is being used to evaluate and optimize other transfer technologies (e.g., electroporation and sonoporation), different delivery reagents, and protocols in a variety of cells in vitro. This work represents the first step in utilizing artificial chromosomes as nonviral vectors for gene therapy. PMID- 11378860 TI - Green fluorescent protein (GFP)-dependent separation of bacterial ghosts from intact cells by FACS. AB - BACKGROUND: E. coli and Salmonella ghost preparations, produced by applying the PhiX174 protein E-mediated lysis system, contain nonlysed bacteria at a very low percentage. To use the ghosts as vaccines, additional methods have to be identified to remove any viable cell, to end up in totally inactivated ghost fractions. Materials and Methods To increase the purity of ghost fractions, we established a green fluorescent protein (GFP)-dependent "in vivo staining" method to be combined with the E-mediated lysis system. Several gfp expression vectors were constructed, and the corresponding cellular fluorescence was analyzed. Bacterial fluorescence, exclusively preserved in nonlysed cells, was utilized to separate these cells from ghost preparations via flow cytometric sorting. RESULTS: High-level production of GFP prior to induction of the lysis system did not affect bacterial growth rates and caused no inhibitory effects on the subsequent protein E-mediated lysis of the cells. The population of reproductive or inactivated but nonlysed cells was highly fluorescent at mean intensities 215 fold higher than ghosts, which exhibited fluorescence at background level. Fluorescent cells could effectively be separated from ghost preparations via flow cytometric sorting. Cell sorting subsequent to protein E-mediated lysis reduced the number of viable cells within ghost preparations by a factor of 3 x 10(5). CONCLUSIONS: The presented procedure is compatible with the protein E-mediated lysis system, is highly effective in separation of nonlysed fluorescent cells, and may serve as a prototype for ghost-purification in applications where only a minimum number of viable cells within ghost preparations can be tolerated. PMID- 11378861 TI - Relationship between chromatin compactness and dye uptake for in situ chromatin stained with DAPI. AB - BACKGROUND: This study investigated the relationship between chromatin compactness, which is directly related to chromatin condensation, and DAPI uptake. Materials and Methods For the structural characterization of in situ chromatin, we used fluorescence microscopy and differential scanning calorimetry on calf thymocytes. The compactness of nuclear chromatin was altered by permeabilizing native cells with NP40 detergent. A time-dependent analysis of detergent effects was performed by acquiring nuclear images at different time intervals after permeabilization. In order to compare nuclei of different sizes, we implemented a geometrical correction in the calculation of the integrated fluorescence intensity. For a quantitative evaluation of chromatin condensation we introduced two new parameters, "average chromatin packing ratio" and "average dye spatial density." RESULTS: This approach allowed us to estimate the effects of NP40 detergent at the level of in situ chromatin. Detergent effects could be modulated by changing the ionic composition of buffer. Moreover, changes of chromatin condensation induced by detergent were inversely related to modifications of nuclear volume. CONCLUSIONS: The combination of complementary information obtained by fluorescence microscopy, supported by a proper geometrical correction, and differential calorimetry allowed us to interpret the patterns of fluorescence intensities inside the nucleus in terms of chromatin structure. PMID- 11378862 TI - A unified procedure for conservative (morphology) and integral (DNA and immunophenotype) cell staining for flow cytometry. AB - BACKGROUND: Current methods for multiparameter DNA flow cytometry suffer from several limitations. These include significant modifications of cell morphological parameters, the impossibility to counterstain cells with certain fluorochromes, and laborious tuning of the instrument that, for some procedures, must be equipped with an ultraviolet (UV) laser. To overcome these problems, we developed a novel method for the simultaneous analysis of morphological parameters, four-color immunophenotyping, and stoichiometric DNA labeling using a bench-top flow cytometer. METHODS: The method consists of a mild permeabilization/fixation treatment at room temperature, followed by labeling with fluorochrome-conjugated monoclonal antibodies (mAbs) and with the DNA dye 7 aminoactinomycin D (7-AAD) at 56 degrees C. RESULTS: Using this method, we analyzed resting peripheral blood mononucleated cells (PBMC), proliferating T cells cultured in the presence of interleukin-2 (IL-2), and lymphoblastoid B cells. Lymphocytes, monocytes, and lymphoblasts treated by this procedure retained differential light scattering (DLS) characteristics virtually identical to those of untreated cells. This allowed regions to be drawn on forward scatter (FSC) and side scatter (SSC) cytograms resolving different cell populations. DLS were preserved well enough to distinguish large lymphoblasts in the S or G2/M phases from small G0/G1 cells. Also, stainability with fluorescein-isothiocyanate (FITC), R-phycoerythrin (PE), allophycocyanin (APC)-conjugated mAbs was generally preserved. DNA labeling with 7-AAD was of quality good enough to permit accurate cell cycle analysis. CONCLUSIONS: The method described here, which we called integral hot staining (IHS), represents a very simple, reproducible, and conservative assay for multiparameter DNA analysis using a bench-top flow cytometer. Last but not least, the cytometer tuning for multiparameter acquisition is straightforward. PMID- 11378863 TI - A simple and highly efficient fixation method for Chrysochromulina polylepis (Prymnesiophytes) for analytical flow cytometry. AB - BACKGROUND: To study the fragile Prymnesiophyte species Chrysochromulina polylepis by flow cytometry (FC), we needed an effective fixation method. This method must guarantee a high yield of fixed cells to achieve acceptable measurement times by FC and to allow quick processing of many samples. Moreover, we wanted a method that allows for storage of fixed samples when FC analysis cannot be done immediately. METHODS: Different aldehydes and methanol were tested at different final concentrations. Gravity sedimentation and centrifugation were applied to achieve higher cell concentrations. Storage of fixed samples was tested under different conditions. RESULTS: 0.25% glutaraldehyde (GA) fixation yielded a recovery rate of about 90%. The signals obtained by FC analysis were excellent. It is possible to centrifuge GA-fixed cells and to store them for several weeks. CONCLUSIONS: GA is the fixative of choice for FC analysis of C. polylepis (and possibly other small delicate species) because it yielded highly significant recovery rates and high-quality FC signals. Cells can be centrifuged to increase the cell concentration, thereby achieving short measurement times with FC. The possibility of long-term storage of fixed cells presents an additional advantage if FC analysis cannot be done immediately. PMID- 11378864 TI - Violet laser diodes as light sources for cytometry. AB - BACKGROUND: Violet laser diodes have recently become commercially available. These devices emit 5-25 mW in the range of 395-415 nm, and are available in systems that incorporate the diodes with collimating optics and regulated power supplies in housing incorporating thermoelectric coolers, which are necessary to maintain stable output. Such systems now cost several thousand dollars, but are expected to drop substantially in price. Materials and Methods A 4-mW, 397-nm violet diode system was used in a laboratory-built flow cytometer to excite fluorescence of DAPI and Hoechst dyes in permeabilized and intact cells. Forward and orthogonal light scattering were also measured. RESULTS: DNA content histograms with good precision (G(0)/G(1) coefficient of variation 1.7%) were obtained with DAPI staining; precision was lower using Hoechst 33342. Hoechst 34580, with an excitation maximum nearer 400 nm, yielded the highest fluorescence intensity, but appeared to decompose after a short time in solution. Scatter signals exhibited relatively broad distributions. CONCLUSIONS: Violet laser diodes are relatively inexpensive, compact, efficient, and quiet light sources for DNA fluorescence measurement using DAPI and Hoechst dyes; they can also excite several other fluorescent probes. PMID- 11378865 TI - Measurement of CD2 expression levels of IFN-alpha-treated fibrosarcomas using cell tracking velocimetry. AB - METHODS: A methodology and a mathematical relationship have been developed that allow quantitation of the expression levels of cellular surface antigens, in terms of antibody binding capacities (ABC). This methodology uses immunomagnetically labeled cells and calibration microbeads combined with cell tracking velocimetry (CTV) technology to measure magnetophoretic mobilities corresponding to cellular ABC. The mobility measurements were accomplished by microscopically recording and calculating the velocity of immunomagnetically labeled QSC microbeads and cells in a nearly constant magnetic energy gradient. RESULTS: Transformed fibrosarcoma cells were given controlled treatments of interferon-alpha in order to manipulate CD2 antigen expression levels. These cells were then immunomagnetically labeled with anti-CD2 FITC antibodies and anti FITC MACS paramagnetic nanoparticles. Measured magnetophoretic mobilities were used to calculate ABC for these cells, corresponding to CD2 expression levels. CONCLUSION: The results from CTV and flow cytometry (FCM) qualitatively verify that these fibrosarcoma cells express elevated levels of CD2 molecules with increasing interferon-alpha treatment from 0 to 24 h. The mean basal CD2 expression level, in terms of ABC, was calculated to be 27,000 from CTV analysis, whereas FCM indicates a comparable ABC value of 33,000. PMID- 11378866 TI - Use of Leu3a/3b for the accurate determination of CD4 subsets for measurement of intracellular cytokines. AB - BACKGROUND: Identification of human T-helper cell subsets is possible by measurement of intracellular cytokines after coincubation of lymphocytes with phorbol myristate acetate (PMA), calcium ionophore, and brefeldin A for up to 20 h. However, exposure to PMA leads to internalization of membrane CD4 and to loss of resolution of the CD4+ cells. Detection of CD3+CD8- cells or preselection of CD4+ cells prior to stimulation is more cumbersome than direct measurement of CD4+ cells. We report the use of the Leu3a/Leu3b multiclone for the accurate determination of CD4 cells after PMA stimulation. METHODS: Peripheral blood lymphocytes were isolated from healthy normal donors and the proportion of CD3+ / CD4+ T cells was determined by flow cytometry before and after incubation with PMA, calcium ionophore, and brefeldin A for 20 h using a variety of anti-CD4 monoclonal antibodies. RESULTS: The Leu3a/3b multiclone reagent was the only anti CD4 monoclonal antibody capable of resolving more than 98% of the initial CD4+ events after incubation with PMA. CONCLUSIONS: The higher signal-to-noise ratio associated with Leu3a3b reagent, compared with other CD4-specific antibodies available, allows the direct and accurate identification of the CD4 subset even after PMA treatment of cells. PMID- 11378867 TI - Automation of mouse micronucleus genotoxicity assay by laser scanning cytometry. AB - BACKGROUND: The evaluation of the safety of drugs and other chemicals is an important aspect of toxicology work. The mouse micronucleus assay is a standard in vivo genotoxicity assay. Chromosomal damage is an indicator of genotoxicity, which manifests in the formation of micronuclei in polychromatic erythrocytes from bone marrow and in peripheral blood erythrocytes. The assay is laborious to perform by manual counting. The laser scanning cytometer allows automated and rapid quantitation of cellular and subcellular fluorescence in monodisperse cell samples on a microscope slide. The object of this study was to evaluate the application of this new technology in the mouse micronucleus genotoxicity assay. Materials and Methods One hundred forty-four mice of various strains were dosed with combinations of carcinogens and antioxidants. Duplicate blood films were prepared 3 days later. One set of slides was stained with acridine orange, and the proportion of micronucleated erythrocytes was counted in 5,000 cells per slide. The duplicates were stained with propidium iodide (40 microg/ml). Five thousand cells per sample were examined using a laser scanning cytometer. The proportion of micronucleated erythrocytes was measured. RESULTS: A coefficient of correlation of 0.96 was found between the data from the two assays. The automation of the assay on the LSC produced a considerable time saving and efficiency gain. CONCLUSIONS: We conclude that with further development, laser scanning cytometry is likely to become the preferred modality for the performance of standard genotoxicity assays. PMID- 11378868 TI - Automated evaluation of frequencies of aneuploid sperm by laser-scanning cytometry (LSC). AB - BACKGROUND: Laser-scanning cytometry (LSC) allows fast automated scoring of fluorescence signals directly on microscopic slides. Frequencies of spontaneous aneuploidies in murine and human sperm were evaluated by using this new LSC technique. Rapid detection may be of great interest in reproductive toxicology, as certain chemicals act as aneugens during meiosis, increasing the production of aneuploid germ cells. Materials and Methods Selected chromosomes were detected by using fluorescence in situ hybridization (FISH) and fluorochrome-labeled DNA probes. Sperm chromatin was counterstained with propidium iodide. By scanning across the slide, fluorescence signals within sperm nuclei were detected and counted. RESULTS: In murine sperm, the frequencies of disomies for chromosomes 8 and X were 0.019% and 0.021%, respectively. The automated assessment in human sperm resulted in disomy frequencies of 0.061% and 0.090% for chromosomes 13 and X, respectively. These results were comparable to data obtained from the same samples by manual microscopic scoring and to literature data. CONCLUSIONS: Frequencies of genotypically abnormal sperm were not significantly different between automated and manual scoring. In conclusion, sperm aneuploidy was reliably determined and disomic sperm were successfully relocated by LSC. By virtue of rapid and reliable analyses, LSC has the powerful potential to replace manual microscopic FISH analysis in molecular cytogenetics. PMID- 11378869 TI - Anisotropy in high angular resolution diffusion-weighted MRI. AB - The diffusion in voxels with multidirectional fibers can be quite complicated and not necessarily well characterized by the standard diffusion tensor model. High angular resolution diffusion-weighted acquisitions have recently been proposed as a method to investigate such voxels, but the reconstruction methods proposed require sophisticated estimation schemes. We present here a simple algorithm for the identification of diffusion anisotropy based upon the variance of the estimated apparent diffusion coefficient (ADC) as a function of measurement direction. The rationale for this method is discussed, and results in normal human subjects acquired with a novel diffusion-weighted stimulated-echo spiral acquisition are presented which distinguish areas of anisotropy that are not apparent in the relative anisotropy maps derived from the standard diffusion tensor model. Published 2001 Wiley-Liss, Inc. PMID- 11378870 TI - Dynamic imaging with multiple resolutions along phase-encode and slice-select dimensions. AB - An implementation is reported of an imaging method to obtain MUltiple Resolutions along Phase-encode and Slice-select dimensions (MURPS), which enables dynamic imaging of focal changes using a graded, multiresolution approach. MURPS allows one to trade spatial resolution in part of the volume for improved temporal resolution in dynamic imaging applications. A unique method of Hadamard slice encoding is used, enabling the varying of the phase encode and slice resolution while maintaining a constant effective TR throughout the entire 3-D volume. MURPS was implemented using a gradient-recalled echo sequence, and its utility was demonstrated for MR temperature monitoring. In this preliminary work, it has been shown that changes throughout a large volume can be effectively monitored in times that would normally only permit dynamic imaging in one or a very few slices. PMID- 11378871 TI - Reliable detection of macromolecules in single-volume 1H NMR spectra of the human brain. AB - In short echo time proton MR spectra of the brain, resonances from macromolecules are visible. The macromolecular resonances in the 0.5-2.0 ppm region can be affected by lipid contamination arising from fat-containing regions outside the selected volume of interest (VOI). This study demonstrates that considerable lipid contamination may remain in stimulated echo acquisition mode (STEAM) spectra even if the spoiling of unwanted coherences is sufficient and the VOI is placed 2 cm or more away from fat-containing regions. The observed contamination was attributed to residual remote out-of-volume excitation, although only very small out-of-slice ripples of less than 0.2% of the in-slice excitation were found in the calculated excitation profile of the RF pulses. Spatial presaturation of fat-containing regions led to a sufficient suppression of the contamination and enabled the detection of highly reproducible macromolecular resonances. Thus, in single-volume spectroscopy as well as in spectroscopic imaging (SI or CSI), the combination of volume selection and outer volume presaturation, each in three dimensions, is highly recommended to ensure accurate detection and reliable evaluation of even small pathological alterations in macromolecules, e.g., proteins or lipids, or other resonances in the 0.5-2.0 ppm region. PMID- 11378872 TI - Response of metabolites with coupled spins to the STEAM sequence. AB - This article demonstrates that a numerical solution of the full quantum mechanical equations for all metabolites with coupled spins is an efficient and accurate means, first, of predicting the optimum STEAM sequence design for quantifying any target metabolite in brain, and, second, for providing the basis lineshapes and yields of these metabolites to facilitate their accurate quantification. Using as illustrations the weakly coupled AX3 system of lactate, the ABX aspartyl group of N-acetylaspartate, which has only two strongly coupled spins, and the much larger strongly coupled AMNPQ glutamyl group of glutamate, the numerical solutions for the response to STEAM highlight the principal source of response variability, namely, the evolution of and transfer between zero quantum terms during the mixing time, TM. These highlights include the rapid oscillations of zero quantum terms due to the chemical shift difference of the coupled spins, the proliferation of oscillating zero order terms due to strong coupling, and the serendipitous smoothing of the response as the number of strongly coupled spins increases. The numerical solutions also demonstrate that the design of the selective 90 degrees pulses is a far less critical factor in determining the response than was the case for the selective 180 degrees pulses of the PRESS sequence (Thompson and Allen, Magn Reson Med 1999;41:1162-1169). The veracity of the method is demonstrated both in phantom solutions and in the parietal lobe of a normal human brain. PMID- 11378873 TI - Representation of strong baseline contributions in 1H MR spectra. AB - A comparison is made between two optimization procedures and two data models for automated analysis of in vivo proton MR spectra of brain, typical of that obtained using MR spectroscopic imaging at 1.5 Tesla. First, a shift invariant wavelet filter is presented that provides improved performance over a conventional wavelet filter method for characterizing smoothly varying baseline signals. Next, two spectral fitting methods are described: an iterative spectral analysis method that alternates between optimizing a parametric description of metabolite signals and nonparametric characterization of baseline contributions, and a single-pass method that optimizes a complete spectral and baseline model. Both methods are evaluated using wavelet and spline models of the baseline function. Results are shown for Monte Carlo simulations of data representative of both long and short TE, in vivo 1H acquisitions. PMID- 11378874 TI - Single-voxel oversampled J-resolved spectroscopy of in vivo human prostate tissue. AB - Single-voxel J-resolved spectroscopy with oversampling in the F1 dimension was used to obtain water unsuppressed 1H spectra of in situ human prostate tissue in 40 previously untreated prostate cancer patients. Based on T2-weighted MRI and previous biopsy information, voxels were placed in regions of benign or malignant peripheral zone tissue, or in regions of predominantly glandular or stromal benign prostatic hyperplasia (BPH) within the central gland. The addition of a second J-resolved dimension allowed for the observation of the J-modulation of citrate, as well as the resolution of polyamines from overlapping choline and creatine signals. Regions of healthy peripheral zone tissue and glandular BPH all demonstrated high levels of citrate and polyamines, with consistent coupling and J-modulation patterns. Conversely, regions of malignant peripheral zone tissue and stromal BPH demonstrated low levels of citrate and polyamines consistent with prior in vivo and ex vivo studies. Moreover, water T2 relaxation times determined for healthy peripheral zone tissue (mean 128 +/- 15.2 msec) were significantly different than for malignant peripheral zone tissue (mean 88.0 +/- 14.2 msec, P = 0.005), as well as for predominantly glandular (mean 92.4 +/- 12.2 msec, P = 0.009) and stromal BPH (mean 70.9 +/- 12.1 msec, P = 0.003). This preliminary study demonstrates that J-resolved spectroscopy of the in situ prostate can be acquired, and the information obtained from the second spectral dimension can provide additional physiologic information from human prostate tissue in a reasonable amount of time (< 10 min). PMID- 11378875 TI - [1-13C]glucose MRS in chronic hepatic encephalopathy in man. AB - [1-13C]-labeled glucose was infused intravenously in a single dose of 0.2 g/kg body weight over 15 min in six patients with chronic hepatic encephalopathy, and three controls. Serial 13C MR spectra of the brain were acquired. Patients exhibited the following characteristics relative to normal controls: 1) Cerebral glutamine concentration was increased (12.6 +/- 3.8 vs. 6.5 +/- 1.9 mmol/kg, P < 0.006) and glutamate was reduced (8.2 +/- 1.0 vs. 9.9 +/- 0.6 mmol/kg, P < 0.02). 2) 13C incorporation into glutamate C4 and C2 positions was reduced in patients (80 min after start of infusion C4: 0.43 +/- 0.09 vs. 0.84 +/- 0.15 mmol/kg, P < 0.001; C2: 0.20 +/- 0.03 vs. 0.45 +/- 0.07 mmol/kg, P < 0.0001). 3) 13C incorporation into bicarbonate was delayed (90 +/- 21 vs. 40 +/- 10 min, P < 0.003), and the time interval between detection of glutamate C4 and C2 labeling was longer in patients (22 +/- 8 vs. 12 +/- 3 min, P < 0.03). 4) Glutamate C2 turnover time was reduced in chronic hepatic encephalopathy (17.1 +/- 6.8 vs. 49.6 +/- 8.7 min, P < 0.0002). 5) 13C accumulation into glutamine C2 relative to its substrate glutamate C2 increased progressively with the severity of clinical symptoms (r = 0.96, P < 0.01). These data indicate disturbed neurotransmitter glutamate/glutamine cycling and reduced glucose oxidation in chronic hepatic encephalopathy. [1-13C] glucose MRS provides novel insights into disease progression and the pathophysiology of chronic hepatic encephalopathy. PMID- 11378876 TI - Detection of an intense resonance at 2.4 ppm in 1H MR spectra of patients with severe late-delayed, radiation-induced brain injuries. AB - Proton MRS and MRI were used to monitor the progression of severe cerebral radiation injuries in 10 patients over a period of 18 months. An unknown resonance (Px) in the 2.37-2.40 ppm region was consistently detected in the affected temporal lobes of four patients. The detection of Px was only confined to spectra with lactate (Lac) and in patients with the highest severity grading of radiation injury. The incidence of Px in Lac-positive spectra was 42.8% (15/35) and in lesions with highest injury grading was 46.8% (15/32). Lesions with Px had significantly higher Lac/creatine (Cr) ratios and more extensive mass effect changes when compared to lesions without Px. The probable identity of Px was examined in the context of anaerobic glycolysis producing pyruvate (2.37 ppm) and the model of metabolic changes in brain abscess formation implicating succinate (2.40 ppm). PMID- 11378877 TI - 1H NMR visible lipids are induced by phosphonium salts and 5-fluorouracil in human breast cancer cells. AB - Cationic lipophilic phosphonium salts (CLPS) selectively accumulate in the mitochondria of neoplastic cells and inhibit mitochondrial function. The effects of the CLPS p-(triphenylphosphoniummethyl) benzaldehyde chloride (drug A), and [4 (hydrazinocarboxy)-1-butyl] tris-(4-dimethylaminophenyl) phosphonium chloride (drug B), on human breast cells of differing biological properties were assessed using growth inhibition assays and 1H NMR. Drug A and, to a lesser extent, drug B demonstrated selective growth inhibition of the highly tumorigenic DU4475 breast carcinoma cell line compared to the transformed HBL-100 human breast cell line. However, in contrast to previous studies using other cell lines, no synergistic activity was found when the drugs were used in combination. 1H NMR demonstrated significant increases in mobile lipid acyl chain resonances in both cell lines treated with cytotoxic doses (IC50, 48 h) of the drugs used either alone or in combination. Two-dimensional NMR revealed accompanying decreases in phosphocholine/Lys levels in HBL-100 cells treated with A, B, or a 1:1 combination A+B at the IC50, and in DU4475 cells treated with drug A (IC50). This was accompanied by significant increases in cho/Lys ratios with IC50 A or combination A+B treatment. Similar spectra were observed in cells treated with 5 fluorouracil but not methotrexate, indicating that mobile lipid accumulation is a general but not universal response to cytotoxic insult. PMID- 11378878 TI - MRI mapping of cerebrovascular reactivity using square wave changes in end-tidal PCO2. AB - Cerebrovascular reactivity can be quantified by correlating blood oxygen level dependent (BOLD) signal intensity with changes in end-tidal partial pressure of carbon dioxide (PCO2). Four 3-min cycles of high and low PCO2 were induced in three subjects, each cycle containing a steady PCO2 level lasting at least 60 sec. The BOLD signal closely followed the end-tidal PCO2. The mean MRI signal intensity difference between high and low PCO2 (i.e., cerebrovascular reactivity) was 4.0 +/- 3.4% for gray matter and 0.0 +/- 2.0% for white matter. This is the first demonstration of the application of a controlled reproducible physiologic stimulus, i.e., alternating steady state levels of PCO2, to the quantification of cerebrovascular reactivity. PMID- 11378879 TI - On T2-shortening by weakly magnetized particles: the chemical exchange model. AB - Chemical exchange (CE) theory is compared with two theories of T2-shortening caused by microscopic magnetic centers: inner- and outer-sphere relaxation theory (long-echo limit) and mean gradient diffusion theory (short-echo limit). The CE equation is shown to be identical to these theories in the respective limits and appropriate parameter relationships are derived for spherical particles. The theories are then compared with computer simulations of spherical particles and with a recent general theory, with good agreement in the asymptotic regions. The CE model also reproduces the essential relaxation characteristics in the intermediate range. Finally, good agreement of a CE model with simulations for magnetized cylinders is also demonstrated. The discussion is limited to weakly magnetized particles such that the maximum phase shift during an echo interval is less than one radian, permitting the use of the Luz-Meiboom CE equation. Published 2001 Wiley-Liss, Inc. PMID- 11378880 TI - Perfusion imaging using dynamic arterial spin labeling (DASL). AB - Recently, a technique based on arterial spin labeling, called dynamic arterial spin labeling (DASL (Magn Reson Med 1999;41:299-308)), has been introduced to measure simultaneously the transit time of the labeled blood from the labeling plane to the exchange site, the longitudinal relaxation time of the tissue, and the perfusion of the tissue. This technique relies on the measurement of the tissue magnetization response to a time varying labeling function. The analysis of the characteristics of the tissue magnetization response (transit time, filling time constant, and perfusion) allows for quantification of the tissue perfusion and for transit time map computations. In the present work, the DASL scheme is used in conjunction with echo planar imaging at 4.7 T to produce brain maps of perfusion and transit time in the anesthetized rat, under graded hypercapnia. The data obtained show the variation of perfusion and transit time as a function of arterial pCO2. Based on the data, CO2 reactivity maps are computed. Published 2001 Wiley-Liss, Inc. PMID- 11378881 TI - Dynamic contrast-enhanced MRI using Gd-DTPA: interindividual variability of the arterial input function and consequences for the assessment of kinetics in tumors. AB - Gd-DTPA kinetics in arterial blood was investigated by dynamic MRI in 47 patients with malignant and benign mammary tumors. Signal enhancement was monitored for 10 min after the beginning of a 1-min infusion of 0.1 mmol/kg Gd-DTPA. Kinetics in blood was biexponential with median half-lives of 21 sec and 11.1 min, respectively. Peak signal enhancement and the area under the signal enhancement time curve varied 2.5- and 3.7-fold between patients. The shortest mean residence time in one of up to three tumor compartments, MRT*, was estimated using either the individual (reference) or a mean population (surrogate) arterial input function (AIF). MRT* (reference estimate) was 1.0 (0-1.5), 1.9 (1.5-2.3), and 2.5 (2.3-2.8) min in carcinomas, fibroadenomas, and mastopathies, respectively (median and interquartile distance). Surrogate estimates were unbiased but differed from the reference estimates 1.5-fold or more in 23% of cases. AIFs should be monitored individually if accurate estimates of individual MRT* are desired. PMID- 11378882 TI - Two-component diffusion tensor MRI of isolated perfused hearts. AB - Nonmonoexponential MR diffusion decay behavior has been observed at high diffusion-weighting strengths for cell aggregates and tissues, including the myocardium; however, implications for myocardial MR diffusion tensor imaging are largely unknown. In this study, a slow-exchange-limit, two-component diffusion tensor model was fitted to diffusion-weighted images obtained in isolated, perfused rat hearts. Results indicate that there are at least two distinct components of anisotropic diffusion, characterized by a "fast" component whose principal diffusivity is comparable to that of the perfusate, and a highly anisotropic "slow" component. It is speculated that the two components correspond to tissue compartments and have a general agreement with the orientations of anisotropy, or fiber orientations, in the myocardium. Moreover, consideration of previous studies of myocardial diffusion suggests that the presently observed fast component may likely be dominated by diffusion in the vascular space, whereas the slow component may include the intracellular and interstitial compartments. The implications of the results for myocardial fiber orientation mapping and limitations of the current two-component model used are also discussed. PMID- 11378883 TI - Differentiation of nonmetastatic and metastatic rodent prostate tumors with high spectral and spatial resolution MRI. AB - MR images can be acquired with high spectral and spatial resolution to precisely measure lineshapes of the water and fat resonances in each image voxel. Previous work suggests that the high-resolution spectral information can be used to improve image contrast, SNR, sensitivity to contrast agents and to physiologic and biochemical processes that affect local magnetic susceptibility gradients. The potential advantages of high-resolution spectroscopic imaging (SI) suggest that it might be useful for early detection and characterization of tumors. The present experiments evaluate the use of high-resolution SI to discriminate between metastatic and nonmetastatic rodent Dunning prostate tumors. SI datasets were obtained at 4.7 Tesla with an in-plane resolution of 350-500 micron in a single 1.0-mm slice, and 6-8 Hz spectral resolution, before and after i.v. injection of an iron oxide contrast agent. Images of water signal peak height in nonmetastatic tumors were smoother in the tumor interior than images of metastatic tumors (P <.004 by t-test) before contrast media injection. This difference was stronger in contrast-enhanced images (P <.0004). In addition, the boundary between the tumor and muscle was more clearly demarcated in nonmetastatic than metastatic tumors. Combinations of image texture, tumor edge morphology, and changes in T2* following contrast media injection improved discrimination between metastatic and nonmetastatic tumors. The data presented here do not demonstrate that effective discrimination between metastatic and nonmetastatic tumors depends on the use of high-resolution SI. However, the results suggest that SI and/or other MR methods that provide similar contrast might be used clinically for early and accurate detection of metastatic disease. PMID- 11378884 TI - Utilizing the diffusion-to-noise ratio to optimize magnetic resonance diffusion tensor acquisition strategies for improving measurements of diffusion anisotropy. AB - It is well known that quantitative anisotropy measurements derived from the diffusion tensor are extremely sensitive to noise contamination. The level of noise in the diffusion tensor imaging (DTI) experiment is usually measured from some estimate of the signal-to-noise ratio (SNR) in the component diffusion weighted (DW) images. This measure is, however, highly dependent on experimental parameters, such as the diffusion attenuation b-value and the diffusion coefficient of the subject. Conversely, the diffusion-to-noise ratio (DNR), defined as the SNR of the calculated diffusion tensor trace map, provides a reliable estimate of noise contamination, which is largely independent of such parameters. In this work it is demonstrated how reliable anisotropy measurements can be obtained using an image acquisition strategy that optimizes the DNR of the DTI experiment. This acquisition scheme is shown to provide noise-independent measurements of typical diffusion anisotropy values found in the human brain. PMID- 11378885 TI - VD-AUTO-SMASH imaging. AB - Recently a self-calibrating SMASH technique, AUTO-SMASH, was described. This technique is based on PPA with RF coil arrays using auto-calibration signals. In AUTO-SMASH, important coil sensitivity information required for successful SMASH reconstruction is obtained during the actual scan using the correlation between undersampled SMASH signal data and additionally sampled calibration signals with appropriate offsets in k-space. However, AUTO-SMASH is susceptible to noise in the acquired data and to imperfect spatial harmonic generation in the underlying coil array. In this work, a new modified type of internal sensitivity calibration, VD-AUTO-SMASH, is proposed. This method uses a VD k-space sampling approach and shows the ability to improve the image quality without significantly increasing the total scan time. This new k-space adapted calibration approach is based on a k-space-dependent density function. In this scheme, fully sampled low spatial frequency data are acquired up to a given cutoff-spatial frequency. Above this frequency, only sparse SMASH-type sampling is performed. On top of the VD approach, advanced fitting routines, which allow an improved extraction of coil weighting factors in the presence of noise, are proposed. It is shown in simulations and in vivo cardiac images that the VD approach significantly increases the potential and flexibility of rapid imaging with AUTO-SMASH. PMID- 11378886 TI - Magnetization preparation during the steady state: fat-saturated 3D TrueFISP. AB - A novel fat saturation scheme is proposed which combines spectral fat saturation with a steady-state 3D true Fast Imaging with Steady Precesson (TrueFISP) sequence. Fat saturation consisted of a conventional frequency-selective excitation pulse surrounded by spoiler gradients. This saturation block was periodically repeated within a continuously running TrueFISP sequence. Except for the fat signals, the steady-state signal formation and the resulting image contrast of TrueFISP was not modified by the periodically inserted fat saturation block. This was achieved by a alpha/2 flip-back pulse before the fat saturation block, which stores the established steady-state transverse magnetization as pure longitudinal magnetization. After fat saturation this longitudinal magnetization was excited by a alpha/2 preparation pulse to continue the TrueFISP acquisition. The resulting images show contrast identical to conventional TrueFISP images, but without the usually very bright fat signals. The short repetition time allows acquisition of a 3D data set of the abdomen within a single breath-hold. PMID- 11378887 TI - Bayesian technique for investigating linearity in event-related BOLD fMRI. AB - Event-related BOLD fMRI data is modeled as a linear time-invariant system. Together with Bayesian inference techniques, a statistical test is developed for rigorously detecting linearity/nonlinearity in the BOLD response system. The test is applied to data collected from eight subjects using an event-related paradigm with a switching checkerboard as the visual stimulus. Analyzed as a group, the results clearly find the response to be nonlinear. When each subject is analyzed individually, however, the results are predominantly nonlinear, but there is some evidence to suggest that there may be a crossover from a linear to a nonlinear regime and vice versa. This could be important when estimating physiological parameters for individuals. Additionally, estimates of the hemodynamic response function and corresponding response were obtained, but there was no consistent appearance of a poststimulus undershoot in the event-related BOLD response. PMID- 11378888 TI - BISTRO: an outer-volume suppression method that tolerates RF field inhomogeneity. AB - A technique is described for performing frequency-selective signal suppression with a high degree of tolerance to RF field inhomogeneity. The method is called B1-insensitive train to obliterate signal (BISTRO). BISTRO consists of multiple amplitude- and frequency-modulated (FM) pulses interleaved with spoiler gradients. BISTRO was developed for the purpose of accomplishing band-selective signal removal, as in water suppression and outer-volume suppression (OVS), in applications requiring the use of an inhomogeneous RF transmitter, such as a surface coil. In the present work, Bloch simulations were used to illustrate the principles and theoretical performance of BISTRO. Its performance for OVS was evaluated experimentally using MRI and spectroscopic imaging of phantoms and in vivo animal and human brain. By using FM pulses featuring offset-independent adiabaticity, BISTRO permitted high-quality, broadband suppression with one (or two) discrete borders demarcating the edge(s) of the suppression band. Simulations and experiments demonstrated the ability to operate BISTRO with reasonably attainable peak RF power levels and with average RF energy deposition similar to other multipulse OVS techniques. PMID- 11378889 TI - SMASH and SENSE: experimental and numerical comparisons. AB - Three parallel-imaging methods were implemented and compared in terms of artifact and noise content: original SMASH, Cartesian SENSE, and an extremely simple method called here the "scissors method." These methods represent very different approaches to the parallel-imaging problem. The experimental and numerical comparisons presented here aim at shedding light on the whole spectrum of parallel-imaging methods, not just the three methods actually implemented. In our results, SMASH images had an artifact level significantly higher than SENSE images for all acceleration factors. The SNR in SENSE images was nearly optimal at low acceleration factors. As acceleration was increased, the noise content in SENSE images eventually sharply departed from optimal values, while the artifact content remained low. PMID- 11378890 TI - Proton magnetic resonance spectroscopy of Sjogren-Larsson syndrome heterozygotes. AB - A deficit of fatty alcohol:NAD+ oxidoreductase complex (FAO) activity has been detected in patients with the Sjogren-Larsson syndrome (SLS). A moderate decrease in FAO activity has also been reported in heterozygote SLS subjects. Abnormal peaks were detected with proton magnetic resonance spectroscopy (1H-MRS) in homozygote SLS subjects. The purpose of this study was to examine whether 1H-MRS can be used to detect metabolic and/or pathological abnormalities in heterozygote SLS subjects. Four SLS heterozygotes were examined using 1H-MRS. A moderate decrease in FAO activity was demonstrated in two of the four heterozygotes. Abnormal peaks were detected at 0.9 ppm in the spectrum from cerebral hemispheres of every heterozygote. 1H-MRS was able to detect an abnormal accumulation of fatty alcohols and lipids, which is expected to increase due to an decrease in FAO activity or dysmyelination in heterozygote SLS subjects. Thus, 1H-MRS is suggested to be a powerful tool in the screening of SLS heterozygotes. PMID- 11378891 TI - Biexponential modeling of multigradient-echo MRI data of the brain. AB - Functional MRI (fMRI) using fast multigradient-echo acquisition methods allows the quantitative determination of the relevant parameter T2*. Previously, the TE dependent signal decay has been modeled with a monoexponential function despite the complex composition of the brain. In this study, biexponential modeling was used to evaluate the relaxation of brain parenchyma and blood separate from that of cerebrospinal fluid. Single-shot multigradient-echo data acquired with spiral or EPI techniques were analyzed. In phantom experiments the biexponential method proved to be accurate. Compared to the biexponential procedure, the monoexponential model overestimated T2* (72.2 msec vs. 65.3 msec) and underestimated DeltaT2* (2.96 msec vs. 3.19 msec) during visual stimulation. The biexponential method may allow intrinsic correction for partial volume effects due to cerebrospinal fluid. The activation-induced parameter changes are detected with a sensitivity equal to that of a monoexponential method. The resulting T2* and DeltaT2* values describe the experimental data more accurately. PMID- 11378892 TI - Magnetic resonance microscopy of morphological alterations in mouse trabecular bone structure under conditions of simulated microgravity. AB - This work describes the use of magnetic resonance (MR) microscopy to examine changes in tibial trabecular bone structure in mice following 28 days of hindlimb suspension, a model simulating the effects of microgravity in rodents. In this first MR study involving mice, analysis of 3D images showed that apparent bone volume fraction, trabecular number, and trabecular thickness were decreased, and apparent trabecular spacing increased, significantly (P < 0.05) in hindlimb suspended mice compared to controls. These changes agreed well with light microscopy measurements from an independent study and also with actual spaceflight experiments with rats. PMID- 11378893 TI - Diffusion time dependence of the apparent diffusion tensor in healthy human brain and white matter disease. AB - The diffusion time dependence of the brain water diffusion tensor provides information regarding diffusion restriction and hindrance but has received little attention, primarily due to limitations in gradient amplitude available on clinical MRI systems, required to achieve short diffusion times. Using new, more powerful gradient hardware, the diffusion time dependence of tensor-derived metrics were studied in human brain in the range 8-80 ms, which encompasses the shortest diffusion times studied to date. There was no evidence for a change in mean diffusivity, fractional anisotropy, or in the eigenvalues with diffusion time in healthy human brain. The findings are consistent with a model of unrestricted, but hindered water diffusion with semipermeable membranes, likely originating from the extracellular space in which the average extracellular separation is less than 7 microns. Similar findings in two multiple sclerosis plaques indicated that the size of the water diffusion space in the lesion did not exceed this dimension. PMID- 11378894 TI - Rat lung MRI using low-temperature prepolarized helium-3. AB - The purpose of this study was to evaluate the recently proposed technique of 3He prepolarization at low temperature and high field (Kober et al. Magn Reson Med 1999; 41:1084-1087) for fast imaging of the lung. Helium-3 was cooled to 2.4 K in a magnetic field of 8 Tesla to obtain a polarization of 0.26%. The polarized 3He was warmed up to room temperature and transferred to a rat, with a final polarization of about 0.1%, large enough for acquiring a 3D image of the rat lung in 30 s. PMID- 11378895 TI - Dependence of venous enhancement on the field of view in 3D contrast-enhanced MRA using the elliptical centric view order. AB - The dependence of venous suppression on the acquisition field of view (FOV) in elliptical centric 3D contrast-enhanced magnetic resonance angiography (CE-MRA) is studied theoretically and experimentally. It is hypothesized that a reduced FOV in an arterial phase acquisition results in improved venous suppression. An expression is derived linking the k-space representation of a vein to venous return time and acquisition parameters. For a y x z FOV reduction from 24 cm x 7.2 cm to 18 cm x 3.6 cm, equivalent voxel size, and venous return times ranging from 0-7 s, the mean improvement in venous suppression ranged from 7.0% for the 19-mm-diameter vein to 32.1% in the 6-mm-diameter vein, assuming a step function shaped venous enhancement profile. Decreased venous enhancement with reduced FOV is also observed for scans with equivalent acquisition times, although the degree of suppression is dependent on the shape of the venous enhancement curve. PMID- 11378896 TI - Sterol glycosides and cerebrosides accumulate in Pichia pastoris, Rhynchosporium secalis and other fungi under normal conditions or under heat shock and ethanol stress. AB - The occurrence of glycolipids such as sterol glycosides, acylated sterol glycosides, cerebrosides and glycosyldiacylglycerols was examined in the three yeast species Candida albicans, Pichia pastoris and Pichia anomala, as well as in the six fungal species Sordaria macrospora, Pyrenophora teres, Ustilago maydis, Acremonium chrysogenum, Penicillium olsonii and Rhynchosporium secalis. Cerebroside was found in all organisms tested, whereas acylated sterol glycosides and glycosyldiacylglycerols were not found in any organism. Sterol glycosides were detected in P. pastoris strain GS115, U. maydis, S. macrospora and R. secalis. This glycolipid occurred in both yeast and filamentous forms of U. maydis but in neither form of C. albicans. This suggests that sterol glycoside is not correlated with the separately grown dimorphic forms of these organisms. Cerebrosides and sterol glycosides from P. pastoris and R. secalis were purified and characterized by mass spectrometry and nuclear magnetic resonance spectroscopy. The cerebrosides are beta-glucosyl ceramides consisting of a saturated alpha-hydroxy or non-hydroxy fatty acid and a Delta4,8-diunsaturated, C9-methyl-branched sphingobase. Sterol glycoside from P. pastoris was identified as ergosterol-beta-D-glucopyranoside, whereas the sterol glucosides from R. secalis contain two derivatives of ergosterol. The biosynthesis of sterol glucoside in P. pastoris CBS7435 and GS115 depended on the culture conditions. The amount of sterol glucoside in cells grown in complete medium was much lower than in cells from minimal medium and a strong increase in the content of sterol glucoside was observed when cells were subjected to stress conditions such as heat shock or increased ethanol concentrations. From these data we suggest that, in addition to Saccharomyces cerevisiae, new yeast and fungal model organisms should be used to study the physiological functions of glycolipids in eukaryotic cells. This suggestion is based on the ubiquitous and frequent occurrence of cerebrosides and sterol glycosides, both of which are rarely detected in S. cerevisiae. We suggest P. pastoris and two plant pathogenic fungi to be selected for this approach. PMID- 11378897 TI - Ypt protein prenylation depends on the interplay among levels of Rab escort protein and geranylgeranyl diphosphate in yeast cells. AB - Farnesyl diphosphate (FPP), an intermediate of the sterol biosynthetic pathway, is used by farnesyl transferase to farnesylate, among others, the Ras proteins, and by geranylgeranyl diphosphate synthase to produce geranylgeranyl diphosphate (GGPP). GGPP is then transferred by geranylgeranyl transferase II (GGTase II) to Rab/Ypt members of the Ras superfamily known to be required at all stages of vesicle transport in both mammals and yeast. Formation of a complex between a Rab/Ypt protein and an accessory protein named the Rab escort protein (REP) is a prerequisite for GGTase II substrate recognition. Little is known about the factors that regulate GGTase II activity in living cells but, based on available data, it seems possible that vesicle transport in higher eukaryotes is regulated by the levels of prenylated Rab/Ypt proteins in the cells. Here we show that the levels of REP play an important role in regulating GGTase II activity in yeast cells if sufficient substrates are present. Moreover, overexpression of REP causes, directly or indirectly, an increased level of Ypt substrates available for prenylation, which in turn leads to the depletion of the GGPP pool in the cell. Overall our data suggest that the levels of REP and the availability of GGPP play a role in regulating Ypt protein prenylation. PMID- 11378899 TI - Constitutive promoter modules for PCR-based gene modification in Saccharomyces cerevisiae. AB - Initial steps in investigating gene function often include deleting and overexpressing the gene of interest and identifying the subcellular location of the gene product. To facilitate these procedures, a number of new PCR modules, which contain selectable markers and in some cases other genetic elements (e.g promoter elements, epitope tags, and reporter genes) have been developed. These modules are used as PCR substrates to create products that can be targeted to specified locations in the yeast genome, thus modifying that genomic locus. We describe here a series of plasmids that contain a truncated version of the strong ADH1 promoter with and without amino-terminal 3HA and GST tags. Because these plasmids contain the same vector sequences as the GAL1 promoter plasmids, a constitutive and an inducible promoter can now be integrated with a minimal number of primers. PMID- 11378898 TI - Sequencing of a 4.3 kbp region of chromosome 2 of Candida albicans reveals the presence of homologues of SHE9 from Saccharomyces cerevisiae and of bacterial phosphatidylinositol-phospholipase C. AB - The nucleotide sequence of a 4.3 kb fragment downstream of the LIG4 gene of Candida albicans has been determined. This fragment contains two entire ORFs (ORF1 and ORF2) and a truncated one (ORF3). ORF1 (1029 bp; EMBL databank, Accession No. AJ277539) encodes a putative protein of 343 amino acids with a high degree of similarity to phosphatidylinositol-specific phospholipases C (PI-PLC) of bacterial origin and, to a lesser degree, to similar proteins from trypanosome, fly and human. Isolated ORF1 confers PI-PLC activity to Escherichia coli transformants. ORF2 (1572 bp; EMBL databank, Accession No. AJ277538) predicts a protein of 524 amino acids with high similarity along most of the entire length to Ydr393w from Saccharomyces cerevisiae. This protein carries a domain with significant similarity to several cytoskeleton proteins of different origins. YDR393w (SHE9) is an orphan gene whose overexpression compromises cell growth. ORF3 appears to encode the homologue of the well-conserved proteasomal 26S regulatory subunit. PMID- 11378900 TI - Isolation of GCR1, a major transcription factor of glycolytic genes in Saccharomyces cerevisiae, from Kluyveromyces lactis. AB - To study the function of GCR1, a gene involved in the expression of glycolytic genes in Saccharomyces cerevisiae, a Kluyveromyces lactis gene that complements the growth defect of a S. cerevisiae Deltagcr1 mutant was isolated. Introduction of this gene into the Deltagcr1 mutant also restored the activities of glycolytic enzymes. DNA sequencing of KlGCR1 predicted an open reading frame of a 767 amino acid protein with an overall identity of 33% and similarity of 48% to Gcr1p from S. cerevisiae. Its DDBJ/EMBL/GenBank Accession No. is AB046391. PMID- 11378901 TI - Cloning of glycerol-3-phosphate dehydrogenase genes (ZrGPD1 and ZrGPD2) and glycerol dehydrogenase genes (ZrGCY1 and ZrGCY2) from the salt-tolerant yeast Zygosaccharomyces rouxii. AB - The ZrGPD1 and ZrGPD2 genes encoding putative glycerol-3-phosphate dehydrogenases were isolated from the salt-tolerant yeast, Zygosaccharomyces rouxii. Both genes are homologous to GPD1 of Saccharomyces cerevisiae and are constitutively expressed in Z. rouxii cells. Putative glycerol dehydrogenase genes, ZrGCY1 and ZrGCY2, which are highly homologous to GCY1 of S. cerevisiae, were also isolated. Since the level of transcripts of ZrGCY1 and ZrGCY2 increased in Z. rouxii cells subjected to salt stress, it is suggested that the pathway of the signal transduction of salt stress controls the expression of these genes. The Accession Nos of these sequences in GenBank are as follows: ZrGPD1, AB047394; ZrGPD2, AB047395; ZrGCY1, AB047396; ZrGCY2, AB047397. PMID- 11378902 TI - Functional characterization of Gms1p/UDP-galactose transporter in Schizosaccharomyces pombe. AB - Galactosylation of glycoproteins in the fission yeast Schizosaccharomyces pombe requires the transport of UDP-galactose as substrate for the galactosyltransferase into the lumen of the Golgi apparatus, which is achieved by the UDP-galactose transporter. We isolated a mutant (gms1) that is deficient in galactosylation of cell surface glycoproteins in Sz.pombe, and found that the gms1(+) gene encodes a UDP-galactose transporter. In the prediction of secondary structure of the Gms1 protein, an eight-membrane-spanning structure was obtained. Fluorescent microscopy revealed the functional Gms1-GFP fusion protein to be stably localized at the Golgi membrane. Sequencing analysis of the coding region of Gms1p derived from galactosylation-defective mutants identified a single amino acid mutation (A102T or A258E) located within the putative transmembrane region, helix 2 or helix 7, respectively. The mutagenized Gms1(A102T or A258E)p exhibited loss of UDP-galactose transport activity but no change in the localization to the Golgi membrane. The C-terminal truncated Gms1p mutants demonstrated that the C terminal hydrophilic region was dispensable for targeting and function as UDP galactose transporter at the Golgi membrane. We suggest that the putative eighth (the most C-terminus-proximal) transmembrane helix of Gms1p is critical to targeting from ER to the Golgi membrane. PMID- 11378904 TI - [The 106th annual meeting of the Japanese Association of Anatomists. Kochi, Japan. April 2-4, 2001. Abstracts]. PMID- 11378903 TI - Identification and characterization of Saccharomyces cerevisiae mutants defective in fluid-phase endocytosis. AB - A mutant library generated by the European Functional Analysis Network (EUROFAN) was screened for strains defective in fluid-phase endocytosis. Accumulation of Lucifer yellow in the vacuole was used as a marker for efficient endocytosis. Fourteen mutants, including ede1Delta, rcy1Delta, sys1Delta and tlg2Delta, previously described to be involved in membrane trafficking, were identified in this screen. alpha-Factor uptake, endocytosis of FM4-64, carboxypeptidase Y secretion, vacuolar morphology, and a vma2 synthetic growth defect were used as criteria to characterize the endocytic defect of the mutant strains obtained. Accordingly, eight mutant strains have endocytic phenotypes in addition to their defect in Lucifer yellow accumulation. These fluid-phase endocytosis mutants are defective at different steps of the endocytic pathway. Interestingly, only two mutants were defective for internalization, two for vacuolar protein sorting and four mutants had aberrant vacuolar morphologies. Some of the mutants identified in this screen that sort carboxypeptidase Y correctly may affect endocytosis at an early post-internalization step before the intersection of the endocytic with the vacuolar protein-sorting pathway. PMID- 11378905 TI - [Morphologic and clinical sequelae of focal ischemic lesions]. AB - The diseases of vessels, mainly of those in brain are one of the most serious problems of the medical practice. The encephalomalacia or cerebral infarctions are usually caused by transient or permanent obstruction of the brain arteries lumen. Beside local dysfunction of vessels the obstructions could be based on embolic events originating in the heart. Such an obstructions are resulting in global and focal cerebral ischaemias. Arterial occlusion results in cerebral ischaemia and the lack of oxygen (anoxia) which leads to reversible or irreversible injury of the nervous cells in the ischaemic region. The local cell injury or cell death causes attraction of macrophages invading into the devitalized tissue within 72-96 hours after the beginning of the ischaemia. The aim of this study was to find out the correlation between asymptomatic or symptomatic course regarding localisation of the ischaemic lesions in the cerebral tissue. Our anatomical findings were collected from 318 autopsies, and reports on postmortem examinations during the period between September-December 1998. The grossing of the brain was carried out by using of Virchow's method. Atherosclerosis, hypertension, and diabetes mellitus were found to be the main risk factors for the production of focal cerebral ischaemia. Of those patients with focal cerebral ischaemia atherosclerosis had 87.5%, 44.3% were suffering from hypertension, and 25% from diabetes mellitus. The focal ischaemia analysed in this study originated from arterial stenosis or thromboembolic obstructions. We divided the lesions into 3 groups according to their size. The most frequently apparent lesions (72%) were the small cysts (0-10 mm in diameter)-lacunae. The majority of them (90%) was found in the basal ganglia. The second group consisted of postmalatic pseudocysts (10-30 mm in diameter), and the third group was represented by encephalomalatic lesions which were larger than 30 mm. Cerebral ischaemic lesions were present in 27.8% of the studied cases. Nevertheless, more than the half (56.8%) of the affected brains (postmalatic pseudocysts, lacunae and malaciae) belongs to the group of patients who were clinically asymptomatic. The asymptomatic lesions, having negative results in the patient's history, and the clinical course were localised mainly in the basal ganglia of both sides and in the frontal part of the right (nondominant) hemisphere. PMID- 11378906 TI - [Detection of silicon in a human spleen using x-ray microanalysis]. AB - There is a lack of information on x-ray microanalysis (XMA) of the human spleen. We have examined the spleen of a patient with autoimmune thrombocytopenic purpura. The specimens were fixed in glutaraldehyde and coated by copper. The XMA was performed by a Quantum detector and a Delta-Class analyzer. In addition to spectral lines of elements normally present in human tissues, we have demonstrated silicon. The presence of silicon in human spleen may be a consequence of the organ's cleansing function. PMID- 11378907 TI - [Multiple intracranial and intraspinal meningiomas in the neurocristopathy (phacomatosis) type of neurofibromatosis]. AB - Presentation of a case of a "central type" neurofibromatosis in a 56-year old woman, clinically diagnosed erroneously as multiple sclerosis with a 20 years long course. Disturbances of hearing, walking, sight, sensitivity, incontinentia, intracranial hypertension and headache represented the main symptoms. More than 120 intracranial and tens of intraspinal meningiomas represented the leading postmortem finding. In a lesser frequency spinal plexiform neurofibromas and schwannomas were also found. The death was attributed to aspiration purulent bronchopneumonia. Various types of meningioma were seen microscopically, including secretory type and a type with amyloid. Immunostaining was positive with S-100 protein and EMA. Negative expression was found with vimentin, CEA, smooth muscle actin, estrogen and progesterone receptors, amyloid A and cytokeratins. With regard to the presence or absence of key morphological features the presented case was placed according to Sobol et al. (29) into the seventh category of neurofibromatosis (NF7). PMID- 11378908 TI - Benign glomus tumor of the superior posterior mediastinum. AB - An unusual location of a benign glomus tumour, outside of the constantly located regions, e.g. in the subungual location or deeply sited in extremities, was diagnosed in a 56-year-old white female in her posterior upper mediastinum. The single similar case report was published before the era of electron microscopy and immunohistochemistry and single cases of atypical and malignant forms in this unusual location were published only recently. The tumour measuring 5 x 4 x 2 centimeters has caused cough and was associated with occasional righ-sided chest pain. Its rich vascular supply has caused intensive intraoperative bleeding. The postoperative course was uneventful and the patient is free of neoplastic disease or symptoms six years after surgery. Numerous mast cells present within the tumour's interstices must be considered in relation to the possible pathogenesis of the up to now unexplained pain in glomus tumours. PMID- 11378909 TI - Composite multicystic mesothelioma and adenomatoid tumor of the ovary: additional observation suggesting common histogenesis of both lesions. AB - An unusual case of composite multicystic mesothelioma and adenomatoid tumor of the ovary is reported. The gradual transitions between both tumor components were well apparent. This observation indicates a histogenetic relationship between adenomatoid tumor and benign multicystic mesothelioma. PMID- 11378910 TI - Kaposiform hemangioendothelioma in adult. Report of a case with amianthoid-like fibrosis and angiectases. AB - A rare case of kaposiform hemangioendothelioma in adult is reported. The 11 x 7 x 5 cm tumor was excised from deep subcutis of the abdominal region in 37-year-old man. No signs of Kasabach-Merritt syndrome or lymphangiomatosis were present. Besides typical pattern of kaposiform hemangioendothelioma, following unusual features were found: dilated vessels producing a gross impression of spindle cell hemangioma, areas of amianthoid-like fibrosis, and diffuse immunoreactivity for CD99. The differential diagnosis included mainly spindle cell hemangioma (hemangioendothelioma), hemangiopericytoma-like solitary fibrous tumor, and Kaposi's sarcoma. PMID- 11378911 TI - [A method of paraffin impregnation and its use in morphologic studies of organs]. AB - The authors present a method of paraffin infiltration applied to heart specimens. They describe the dissection, trimming and fixation of the specimen which is followed by common tissue dehydration and infiltration by paraffin wax. Thus preserved permanent specimen allows both retaining the entire organ and visualizing its particular structures. PMID- 11378912 TI - [Comparison of the results of a medical evaluation of blood alcohol level and other findings]. AB - Evaluation of a relatively large group (3172 subjects) in particular as regards agreement of results of medical examination with the alcohol blood level confirmed the correlation between the behaviour of the examined subjects and alcohol. We expected however that the results will be more unequivocal. It was found that at alcohol blood levels of ca 0.7 g/kg the ratio of examined subjects who manifest or do not manifest disorders of psychosensorimotor function is roughly balanced. From this ensues the necessity to pay more attention to medical examination for further use of the outcome of the examination e.g. for forensic purposes. PMID- 11378913 TI - [Effect of Stopangin mouth spray on blood alcohol levels measured by the Alcotest 7410 analyser made by Drager]. AB - During a check-up of a driver by a police patrol in the Czech Republic by means of a breath analyzer of Drager Co. an alcohol blood level of 0.50 g/kg was assessed. Later the driver reported that before the test he used at 30- and 5 minute intervals the drug Stopangin spray. In the Institute of Forensic Medicine the authors made an experiment which revealed that 22 minutes after administration of the drug the apparatus gave a negative result. Agreement with these conclusions was expressed also by a representative of Drager Co. and the results were published as an expert opinion with a recommended procedure for the police of the Czech Republic in the South Moravian region. PMID- 11378914 TI - What's the Journal of Environmental Radioactivity up to? PMID- 11378915 TI - Investigation of the potential for radon mitigation by operation of mechanical systems affecting indoor air. AB - Mitigation of radon gas and radon progeny in buildings is based largely on reducing the pressure difference between the point of the radiation source and the point of entry to indoor air. This study identifies the influence of mechanical systems, of air-conditioning and 'wet' systems of central heating as potential remediation agents in the control of radon and progeny concentrations. Air-conditioning was found to reduce radon levels in a systematic way within a few hours of start-up, to a low fraction of the immediately preceding concentration. Central heating reduced the level by around 40% of the preceding high within a few hours of start-up. Importantly for health concerns, under operating conditions of both types of system the level of radon progeny was reduced to a greater extent than the radon progenitor. PMID- 11378916 TI - Bronchial Rn dose survey for residences. AB - The bronchial dosimeter for Rn progeny proposed by Yu and Guan in 1998 was employed to survey the bronchial dose from Rn progeny in 30 residences in Hong Kong. An average bronchial deposition fraction of Rn progeny was obtained as 0.0334, which gave an average dose conversion factor (DCF) of 8.5 mSv WLM-1. The mean values of potential alpha energy concentration (PAEC) deposited in the tracheobronchial region (PAECT-B), total PAEC in air (PAECT), annual effective dose (E), concentration of Rn gas (RC) and annual dose conversion factor (ADCF) for all the residential sites combined were 0.11 +/- 0.05, 3.1 +/- 1.4 mWL, 1.2 +/- 0.5 mSv yr-1, 23 +/- 10 Bq m-3 and 0.055 +/- 0.020 (mSv yr-1 per Bqm-3), respectively, with air-conditioned sites (AC sites) and non-AC sites having significantly different mean ADCF values. The indoor relative humidity affected PAECT and RC with high confidence levels (> 95%). PMID- 11378918 TI - Uncertainty analysis of the external gamma-dose rate due to the variability of the vertical distribution of 137Cs in the soil. AB - The external gamma-dose rate at 1 m height above a flat area due to the presence of fallout radiocesium in the soil is frequently calculated from the observed depth profile of the 137Cs activity as well as the soil mass per unit area. At a given site, these depth profiles may, however, vary considerably, thus introducing an uncertainty to the external gamma-dose calculated in this way. To assess this source of uncertainty for a typical grassland site, the activity of Chernobyl-derived 137Cs and the wet bulk density in the three upper soil layers at 100 plots in a 100 m x 100 m pasture were determined. Analysis of these data shows that the frequency distribution of the dose rates calculated from the corresponding depth profiles of all plots is similar to a log-normal distribution (mean 25 nGy h-1, median 22 nGy h-1, standard deviation 11 nGy h-1; range 1.6-56 nGy h-1). The various sources which contribute to the uncertainty of the dose rate are quantified. The semi-variogram indicates that any spatial dependence of the dose rates occurs on this pasture only over distances that are smaller than the shortest sampling interval (here about 10 m). It is estimated which errors have to be expected for the median dose rate when the depth profiles of 137Cs and of the wet bulk density are determined only for a small number of plots. It is preferable to calculate the mean dose rate as a mean from the n individual dose rates rather than from an averaged 137Cs depth profile of the n plots. PMID- 11378917 TI - Natural radioactivity in phosphates, phosphogypsum and natural waters in Morocco. AB - The contents of natural radionuclides (uranium, actinium and thorium series) were measured in sedimentary phosphate rock samples using high-resolution gamma spectrometry. Data obtained for uranium content (ppm) were compared with the results obtained by a method based on the measurements using solid-state nuclear track detectors (SSNTD) in the same samples. The potential leaching of radionuclides from sedimentary phosphate rock during the industrial production of the phosphoric acid was studied. The process of leaching of the radioisotopes from phosphogypsum was discussed. A method for the direct alpha counting of 226Ra thin source, elaborated by the deposition of Ra from aqueous solutions on manganese oxides film deposited on polyvinyl support, have been developed and applied for the determination of 226Ra in natural water samples. The results show that only the water sample from the mine area reveals the presence of 226Ra at a level of about 0.2 Bq l-1. PMID- 11378919 TI - Chemical availability of 137Cs and 90Sr in undisturbed lysimeter soils maintained under controlled and close-to-real conditions. AB - Chemical availability of 137Cs and 90Sr was determined in four undisturbed soils in a lysimeter study three and four years after deposition to the soil surface. The study was part of a larger project on radionuclide soil-plant interactions under well-defined conditions. The soil types were loam, silt loam, sandy loam and loamy sand, and were representatives of important European soil and climatic conditions. The lysimeters were installed in greenhouses with climatic and hydrological control, and were contaminated with 137Cs and 90Sr in an aerosol mixture simulating fallout from a nuclear accident. Soil samples were taken from several depths in each soil in 1997 and 1998 and the samples were sequentially extracted with H2O, NH4Ac, NH2OH.HCl, H2O2 and HNO3. Extractability of 137Cs decreased in the order: HNO3 > R-esidual > or = NH4Ac > H2O2 > or = NH2OH.HCl > or = H2O. More than 80% was found in the acid digestible or residual fractions, and 11-17% in labile fractions. Soil type differences were small. Extractability of 90Sr decreased in the order: NH4Ac > NH2OH.HCl > HNO3 > H2O2 approximately H2O. 31-58% was found in easily available fractions. Differences between soil types were quite small. The results suggest that availability of 137Cs for plant uptake and migration is low, whereas availability of 90Sr is rather high. PMID- 11378920 TI - Fallout strontium and caesium transfer from vegetation to cow milk at two lowland and two Alpine pastures. AB - Vegetation-to-milk transfer coefficients for 137Cs, 90Sr and stable strontium were determined for cows grazing on two intensively managed lowland pastures and two Alpine pastures in Austria. The 90Sr transfer coefficient at the four pastures ranged from 0.0005 to 0.0012 dl-1 and correlated with the stable strontium transfer coefficient (0.0006-0.0013 dl-1) with the lower values found on the intensively managed pastures. The 137Cs transfer coefficient ranged from 0.0009-0.0045 dl-1. PMID- 11378921 TI - Discharge of 137Cs and 90Sr by Finnish rivers to the Baltic Sea in 1986-1996. AB - The total amounts of 137Cs and 90Sr transported from Finland by rivers into the Gulf of Finland, Gulf of Bothnia and Archipelago Sea since 1986 were estimated. The estimates were based on long-term monitoring of 137Cs and 90Sr in river and other surface waters and on the statistics of water discharges from Finnish rivers to the above sub-areas of the Baltic Sea. The total amounts of 137Cs and 90Sr removed from Finland into the Baltic Sea during 1986-1996 were estimated to be 65 and 10 TBq, respectively. The results show that, although the deposition of 137Cs was much higher than that of 90Sr after the Chernobyl accident, the amount of 137Cs removed from Finland is only six times as high as that of 90Sr. This emphasizes the importance of 90Sr while considering radiation doses from surface waters and 137Cs while estimating doses via pathways from catchment soil, lake sediments and biota after a fallout situation. PMID- 11378922 TI - Occurrence of 222Rn, 226Ra, 228Ra and U in groundwater in Fujian Province, China. AB - 222Rn, 226Ra, 228Ra and U were determined in a total of 552 groundwater samples collected throughout Fujian Province of China. The geometric mean concentrations of 222Rn, 226Ra, 228Ra and total U in the groundwater were 147.8 kBq m-3, 12.7 Bq m-3, 30.2 Bq m-3 and 0.54 microgram kg-1, respectively. High groundwater 222Rn was explained by the predominantly granitic rock aquifers in Fujian. A lifetime risk of 1.7 x 10(-3) was estimated for the ingestion of groundwater 222Rn. High ratios of 228Ra to 226Ra contents (geometric mean of 2.4) and their disproportion suggest that 228Ra should also be measured in the assessment of population doses from drinking water in the regions of high rock or soil 232Th. No significant correlation between the 222Rn concentrations in groundwater and air was found. PMID- 11378923 TI - 137Cs and 90Sr behavioural regularities in the southeastern part of the Baltic Sea. AB - Variations in 137Cs concentrations were investigated over the period 1986-1997 in the southeastern part of the Baltic Sea, following the Chernobyl power plant accident. The rate of "self-cleaning" was demonstrated to be very slow, the average concentration of 137Cs in 1996 being almost the same as that measured directly after the accident, in 1986. Measurements of both 137Cs and 90Sr concentrations generally revealed homogeneous distributions in this region of the Baltic Sea, though patchy distributions did develop under some hydrometeorological conditions. Specifically, the 137Cs concentration distribution became heterogeneous with values varying in the range 60-92 Bq/m3 under south-southwesterly wind conditions whilst the 90Sr concentration distribution developed similar characteristics with values ranging from 15 to 64 Bq/m3 under east-southeasterly wind conditions. In addition, in coastal waters, over extensive periods of north-northwesterly winds in 1995, 137Cs concentrations increased to values 1.5-2 times the overall average concentration, which was registered in 1986 and 1996. These data therefore reveal a continuing significant pollution of the waters of the Baltic Sea resulting from the Chernobyl power plant accident, a pollution compounded by the slow rate of radionuclide self cleaning and significant probability of sudden regional concentration increase. PMID- 11378924 TI - Are radioecologists the dinosaurs of this century? PMID- 11378925 TI - Can 239 + 240Pu replace 137Cs as an erosion tracer in agricultural landscapes contaminated with Chernobyl fallout? AB - Erosion studies often use 137Cs from the global fallout (main period: 1953-1964) as a tracer in the soil. In many European countries, where 137Cs was deposited in considerable amounts also by the Chernobyl fallout in 1986, the global fallout fraction (GF-Cs) has to be separated from the Chernobyl fraction by means of the isotope 134Cs. In a few years, this will no longer be possible due to the short half-life of 134Cs (2 yr). Because GF-Cs in the soil can then no longer be determined, the potential of using 239 + 240Pu as a tracer is evaluated. This radionuclide originates in most European countries essentially only from the global fallout. The activities and spatial distributions of Pu and GF-Cs were compared in the soil of a steep field (inclination about 20%, area ca. 3 ha, main soil type Dystric Eutrochrept), sampled at 48 nodes of a 25 x 25 m2 grid. The reference values were determined at 12 points adjacent to the field. Their validity was assured by an inventory study of radiocaesium in a 70 ha area surrounding the field sampling 275 nodes of a 50 x 50 m2 grid. In the field studied, the activity concentrations of GF-Cs and Pu in the Ap horizon were not correlated (Spearman correlation coefficient R = 0.20, p > 0.05), and the activity balance of Pu differed from that of GF-Cs. Whereas no net loss of GF-Cs from the field was observed as compared to the reference site, Pu was more mobile with an average loss of ca. 11% per unit area. In addition, the spatial pattern of GF-Cs and Pu in the field differed significantly. The reason may be that due to their different associations with soil constituents, Pu and Cs represent different fractions of the soil, exhibiting different properties with respect to erosion/deposition processes. This indicates that both radionuclides or one of them may not be appropriate to quantity past erosion. When tracer losses are used to calibrate or verify erosion prediction models, systematic deviations may not only stem from model shortcomings but also from tracer technique. PMID- 11378926 TI - The future of radioecology: in partnership with chemo-ecology and eco-ethics. PMID- 11378927 TI - Geographical mapping and associated fractal analysis of the long-lived Chernobyl fallout radionuclides in Greece. AB - Immediately after the Chernobyl accident, a soil sampling programme was undertaken in order to detect and quantitatively analyse the long-lived radionuclides in the Chernobyl fallout. Soil samples (1242 in number) of 1 cm thick surface soil were collected in Greece during the period from May-November 1986. The samples were counted and analysed using Ge detector set-ups. The 137Cs fallout data have already been analysed, mapped and published. In an attempt to improve this analysis and also to extent it to other fallout radionuclides, an in house unix-based data base/geographical information system (DBGIS) was developed. Multifractal analyses of the deposition patterns have also been performed. In the present work, an analysis of the results of the deposition of 137Cs, 134Cs, 144Ce, 141Ce, 125Sb, 110mAg, 106Ru, 103Ru, 95Zr and 54Mn are presented together with relevant fractal analysis and three characteristic contour maps. The maximum detected values of the above-mentioned radionuclides were 149.5 +/- 0.1, 76.1 +/- 0.1, 32.9 +/- 0.2, 46 +/- 2, 4.56 +/- 0.02, 7.98 +/- 0.02, 79.1 +/- 0.4, 337 +/- 2, 20.1 +/- 0.2 and 3.02 +/- 0.02 kBq m-2, respectively. Furthermore, a statistical technique to compare contour maps was introduced and applied to explain the differences which appeared in the maps of the above-mentioned radionuclides. PMID- 11378928 TI - Radioactivity in tobacco leaves. AB - The radioactivity in tobacco leaves collected from 15 different regions of Greece before cigarette production was studied in order to find any association between the uptake of the naturally occurring radionuclides and the isotopes of cesium of Chernobyl origin. The activities of the isotopes of radium, 226Ra and 228Ra, in the tobacco leaves reflected their origin from the soil by root uptake rather than from fertilizers used in the tobacco cultivation. Lead-210 originated from the air and was deposited onto the tobacco leaves and trapped by the trichomes. Potassium-40 in the tobacco leaves was due to root uptake either from soil or from fertilizer. The isotopes of cesium, 137Cs and 134Cs, in the tobacco leaves were due to root uptake and not due to deposition onto the leaf foliage as they still remained in soil four years after the Chernobyl reactor accident but were absent from the atmosphere in rain washout (precipitation) and gravitational settling. PMID- 11378929 TI - Radon (222Rn) level variations on a regional scale: influence of the basement trace element (U, Th) geochemistry on radon exhalation rates. AB - The approach proposed in this study provides insight into the influence of the basement geochemistry on the spatial distribution of radon (222Rn) levels both at the soil/atmosphere interface and in the atmosphere. We combine different types of in situ radon measurements and a geochemical classification of the lithologies, based on 1/50,000 geological maps, and on their trace element (U, Th) contents. The advantages of this approach are validated by a survey of a stable basement area of Hercynian age, located in South Brittany (western France) and characterized by metamorphic rocks and granitoids displaying a wide range of uranium contents. The radon source-term of the lithologies, their uranium content, is most likely to be the primary parameter which controls the radon concentrations in the outdoor environment. Indeed, the highest radon levels (> or = 100 Bq m-3 in the atmosphere, > or = 100 mBq m-2 s-1 at the surface of the soil) are mostly observed on lithologies whose mean uranium content can exceed 8 ppm and which correspond to peraluminous leucogranites or metagranitoids derived from uraniferous granitoids. PMID- 11378930 TI - Size-fractionated plutonium isotopes in a coastal environment. AB - We have examined the distribution of individual Pu isotopes (239Pu, 240Pu, and 241Pu) in seawater from the Gulf of Maine (GOM). Samples were size-fractionated with a 1 kD cross-flow ultrafiltration (CFF) membrane. Subfractioned samples were radiochemically purified and Pu isotopes were analyzed using a three-stage thermal ionization mass spectrometer (TIMS). To our knowledge, this is the first time that both size class and Pu isotopic data have been obtained for seawater samples. Within measurement uncertainties a single 240Pu/239Pu atom ratio of 0.18 was found for all sample collection depths and sample size fractions. This signifies a current, single Pu source in GOM waters, namely global fallout, and suggests that no measurable isotopic fractionation occurred during CFF processing. The majority of Pu was found in the low molecular weight fraction (< 1 kD). Colloidal Pu varied from 8% of the total in surface waters to < 1% in the deepest (250 m) seawater sample. Evidence suggests that the vertical distribution of Pu in GOM is primarily controlled by conservative mixing processes. The high Pu fraction found in the low molecular size fraction implies that most of the Pu is in the non-particle-reactive oxidized fraction, and is consistent with the conservative Pu behavior. The activity levels are in agreement with other studies which show a slow decrease in Pu with time due to continued mixing and relatively slow particle removal. PMID- 11378931 TI - Radon exhalation from granites used in Saudi Arabia. AB - Measurements of radon exhalation for a total of 50 selected samples of construction materials used in Saudi Arabia were taken using a radon gas analyzer. These materials included sand, aggregate, cement, gypsum, hydrated lime, ceramics and granite. It was found that the granite samples were the main source of radon emanations. A total of 32 local and imported granite samples were tested. It was found that the radon exhalation rates per unit area from these granite samples varied from not detectable to 10.6 Bq m-2 h-1 with an average of 1.3 Bq m-2 h-1. The linear correlation coefficient between emanated radon and radium content was 0.92. The normalized radon exhalation rates from 2.0 cm thick granite samples varied from not detectable to 0.068 (Bq m-2 h-1)/(Bq kg-1) with an average of 0.030 (Bq m-2 h-1)/(Bq kg-1). The average radon emanation of the granite samples was found to be 21% of the total radium concentration. Therefore, granite can be a source of indoor radon as well as external gamma-radiation from the uranium decay series. PMID- 11378932 TI - Natural radionuclides in drinking water supplies of Sao Paulo State, Brazil and consequent population doses. AB - The activity concentrations of 228Ra, 226Ra and 222Rn have been analysed in 452 drinking water supplies of Sao Paulo State. This study started in 1994 and covered 54% of the 574 existing counties. Concentrations up to 235 and 131 mBq l 1 were observed for 226Ra and 228Ra, respectively, whereas 222Rn concentrations reached 315 Bq l-1. Radiation doses up to 0.3, 0.6 and 3.2 mSv yr-1 were estimated for the critical organs, for the ingestion of 226Ra, 226Ra and 222Rn, respectively. The corresponding committed effective doses reached values of 6 x 10(-3), 2 x 10(-2) and 3 x 10(-1) mSv yr-1, for the same radionuclides. These results indicate that 222Rn makes the highest contribution to the total effective dose. PMID- 11378933 TI - So, what is environmental radioactivity anyway? PMID- 11378934 TI - Describing the observed vertical transport of radiocesium in specific soils with three time-dependent models. AB - In this paper three transport models are applied to describe the vertical migration process of 137Cs in undisturbed soil. The same experimental data set is used for the calibration of each model. These data were obtained from areas in northeast Italy, where the 137Cs deposition following the Chernobyl accident ranged from 20 to 40 kBq m-2. A diffusion model (called DSF) is presented, which takes into account three states of cesium in the soil matrix: mobile, sorbed and bound. It is shown that this model provides a reasonably good description of the temporal evolution of the radiocesium profile in soil. In addition, a new version of the RABES model (called RABES I) is proposed, with only two free parameters: the rate of change of the vertical profile and the diffusion length in the equilibrium. This paper includes a critical evaluation among the DSF, RABES and RABES I models, considering, mainly, their predictive aspect about the temporal change of the radiocesium profile in soil. PMID- 11378935 TI - Studies of the radon concentration in drinking water from the horst Soderasen in southern Sweden. AB - The radon activity concentration in ground water from drilled and dug wells on the horst Soderasen in Southern Sweden has been determined with two different methods, gamma activity measurements with a germanium HPGe detector and alpha activity measurements with plastic track detectors. The results are consistent. High activity concentration is connected to granite bedrock. Dug wells have low concentrations and no trivial correlation between concentration and depth of the well is found. Large local variations exist. Activity concentrations > 700 Bq/l appear to be associated with leakage from layers of volcanic origin. The concentration from drilled wells is found to be quite constant over a 3 year period but short time variations appear to be significant. Evaporation from the open surface of a normal cooking vessel is slow with an activity gradient delta A/A of about 0.1-0.2 per hour at room temperature whereas even modest heating of water in e.g. a coffee machine is very efficient and reduces the radon activity concentration by > 90% in one process. PMID- 11378936 TI - Natural radionuclides in the aquatic environment of a phosphogypsum disposal area. AB - Rock phosphate ore processing and disposal of phosphogypsum contribute to enhanced levels of natural radionuclides in the environment. Studies on the distribution of U-series nuclides were carried out in the Chitrapuzha River, near Cochin, in the southern part of India. The concentrations of radionuclides, especially 226Ra, in the river waters showed enhancement by an order of magnitude relative to the levels in nearby water bodies. The concentrations were influenced by seasonal changes in the river flows during monsoon and summer periods. Ingestion doses via fish and milk have an upper estimate of 18 microSv for the critical population. PMID- 11378937 TI - Study of 222Rn concentrations in drinking water in the north-eastern hydroregions of Poland. AB - This paper presents the results of radon concentration measurements in the drinking water from the municipal water supply system and private wells located in the north-eastern part of Poland. The measurements were carried out on 643 samples using a liquid scintillation method. The mean value was found to be 5262 Bq m-3 with a maximum of 38347 Bq m-3. The samples were obtained from different water-bearing levels, i.e. surface water, deep borehole water and well water and have respective mean values of 3398, 5178 and 6155 Bq m-3. In 57 water samples, a guideline maximum level of 11,000 Bq m-3 was found to be exceeded. The observed radon concentration in water can contribute to a 2% increase in indoor radon concentration. PMID- 11378938 TI - Natural radionuclides in Brazilian mineral water and consequent doses to the population. AB - Based on production data published by the Brazilian Mineral Water Producers Association, bottled mineral waters were bought at the market, covering, at least, 50% of the annual production of each Brazilian geographical region. 228Ra, 226Ra and 210Pb were analyzed and the weighted mean values obtained for Brazilian mineral waters were 0.097, 0.027 and 0.066 Bq l-1 respectively. Individual effective dose rate estimates due to ingestion of mineral water in Brazil were of the order of 10-100 microSv yr-1, depending on the geographical region. Based on the available data, it was also possible to evaluate the per caput effective dose rate and the highest value obtained was 4.3 microSv yr-1 for the northern region. PMID- 11378939 TI - A study of the technologically modified sources of 222Rn and its environmental impact in an Indian U mineralised belt. AB - These investigations present qualitative and quantitative assessments of the 222Rn contributing potential of the technologically modified sources, e.g. mine exhausts and tailings piles, arising due to the mining and processing of uraniferous minerals at Jaduguda, India. The overall geometric mean of the 222Rn flux from low-grade uranium (U) mill tailings being 1.19 Bq m-2 s-1, requisite attenuation of 37.8% to the recommended limit of 0.74 Bq m-2 s-1 can be attained by a typical clay coverage of 20 cm. Theoretically, feasible options for selection of overburden materials for stabilisation of the tailings piles are examined. Extrapolation studies with a turbulent diffusion model yield an atmospheric dispersion pattern of 222Rn in reasonable correspondence with the prevailing concentrations, especially for locations beyond a 4 km radius. The overall 222Rn contribution of the U complex to the ambient air is the same as that contributed by the adjoining land mass of 2.75 km radius in this mineralised terrain. PMID- 11378942 TI - Uranium decay products found on Mir space blanket mitt. AB - The space blanket mitt which covered the Trek detector on Mir during four years of orbital flight has been measured for gamma radiation with HPGe and multidimensional spectrometers. Difference spectra from very-long-period spectrometer runs on the mitt and on a similar non-deployed mitt from the same manufacturer show that the mitt has acquired small but significant amounts of gamma radioactivity during orbital flight. Twelve gamma-ray peaks have been measured in the difference spectra, including peaks identified as due to 214Bi and 214Pb from the uranium-radium alpha decay series, and others possibly due to the uranium-actinium series. This implies the presence of a sparse population of uranium decay products in lower orbital space which can only have come from nuclear explosions, burned-up satellite nuclear batteries, the solar wind, or supernova fragments in the local interstellar medium. PMID- 11378940 TI - Radioactive contamination of tropical rainforest soils in Southern Costa Rica. AB - Radionuclide content in soils from four locations in a tropical rainforest near Golfito in Southern Costa Rica was investigated. For comparison, two nearby locations in open grassland were also studied. From each site 5 soil cores down to a depth of 15 cm were taken. The median contamination with 137Cs was 584 Bq m 2 (reference date 1 January 1996) and the coefficient of variation (CV) was 50%. This contamination can be attributed to global fallout from atmospheric nuclear weapon tests between 1945 and 1980. The mean contamination is slightly lower than the value expected for the latitude (8 degrees 42': 700 Bq m-2), which may be explained by migration of radiocaesium to subsoil below 15 cm or by uptake into the living biomass. Out of the total variability of 50%, around 20% can be attributed to the sampling and measuring process uncertainties, thus leaving a 45% contribution of spatial variability. A significant difference between forest and meadow sites could be detected: the meadow sites showed lower radiocaesium soil inventories (median: 291 Bq m-2) than the forest sites (643 Bq m-2). This may be explained by the agricultural activities carried out on meadow sites which lead to an increased redistribution of caesium in the soil profile and therefore a larger fraction of the total 137Cs lying below 15 cm. Another reason for higher contamination levels under forest can be attributed to the high interception potential of dense tree canopies for dry deposition. Extrapolating the 137Cs concentration below the sampling horizon, i.e. accounting for the cut-off of the profiles by the sampling technique, results in an estimated mean of 710 Bq m-2 for the forest sites, which is very close to the expected figure. The mainly mineral part of the forest soil profiles was analysed for the 137Cs transport parameters, apparent convection velocity (v = 0.14 +/- 0.09 cm a-1) and apparent diffusion constant (D = 0.79 +/- 0.49 cm2 a-1). The maximum concentration can be found at 5.3 +/- 2.9 cm depth, the half-value depth being 7.4 +/- 1.3 cm. The mean 40K activity concentration was 175 Bq kg-1 dry matter (CV = 69%) and 226Ra and 228Ra concentrations of 9.90 Bq kg-1 (CV = 23%) and 7.93 Bq kg-1 (CV = 20%) have been found, respectively. PMID- 11378941 TI - Experimental studies on the deposition to crops of radioactive gases released from gas-cooled reactors--III. Carbon-14 dioxide. AB - Experimental studies were performed under controlled environmental conditions to determine the deposition and subsequent allocation of carbon-14, following a short-term fumigation of major British crops with 14CO2. The deposition velocities varied between 0.075 and 1.32 cm3 g-1 s-1 and 0.0008 and 0.157 cm s-1 on weight and area bases, respectively. The primary sites of deposition were leaves, although other organs, e.g. ears and pods, were increasingly important as the crops developed. Activity was allocated post-deposition to those crop components actively growing at the time of deposition. The allocation was generally completed 24 h after fumigation, the exceptions to this being those crops where reproductive organs developed after the fumigation. A single exponential function was the best fit for the loss of activity from the crops. The data are discussed in the context of modelling carbon-14 contamination of crops following radioactive discharges. PMID- 11378943 TI - Factors controlling 7Be and 210Pb atmospheric deposition as revealed by sampling individual rain events in the region of Geneva, Switzerland. AB - Bulk atmospheric deposition of 7Be and 210Pb has been measured at Versoix, close to Geneva, Switzerland. Collectors were continuously deployed from November 1997 through November 1998 for periods from 1 to 22 days depending on the frequency of rain. The activities of 7Be and 210Pb integrated over the sampling interval were moderately well correlated with rainfall (r2 of 0.66 and 0.55, respectively; p < 0.001) and well correlated one with the other (r2 of 0.91; p < 0.001). The 7Be/210Pb activity ratio is close to 13.5, except for samples collected in the periods of very low precipitation which have a distinctly lower ratio. A simple model accounting for rainfall, seasonal variations and reload of the local atmosphere after a rain event explains 90% of the variance of 7Be and 210Pb deposition. Concentrations of 210Pb and Ca++ in rain were correlated with transport time of air masses over the continent as indicated by reconstruction of air mass trajectories over three days. PMID- 11378944 TI - Effect of pH on the sorption of uranium in soils. AB - This work was undertaken to study the influence of soil type and chemical composition on uranium sorption ratios (SR in 1 kg-1) in order to reduce the uncertainty associated with this parameter in risk assessment models. Thirteen soil samples were collected from three different locations in France under different geological conditions. Clay content varied from 7.0 to 50.0%, pH ranged from 5.5 to 8.8 and organic matter content from 1.0 to 4.6%. Soils were incubated at room temperature in polyethylene packets for 28 days in the presence of 1 mg U kg-1 soil. Sorption ratio values varied from 0.9 to 3198 for all soils with no significant effect of soil texture or of organic matter. However, soil pH was highly linearly correlated with (log SR) as a probable consequence of the existence of different uranium complexes as a function of soil pH. The sorption behaviour differences between UO2(2+) and UO2(2+)-carbonate complexes are so great that any other effect of soil properties on U sorption is hidden. Thus, soil pH should be the focus variable for reduction of the uncertainty associated with the soil Kd value used in environmental risk assessments, even for reducing the uncertainty in site-specific Kd values. PMID- 11378945 TI - Communicating nursing care and crossing language barriers. PMID- 11378946 TI - The ideal state for perioperative nursing. AB - For a typical surgical procedure, a minimum of three individuals document the care provided during the intraoperative phase. This makes it difficult to develop perioperative records with pertinent data elements without creating redundancy, errors, and inconsistencies. This article discusses strategies to develop surgical records that share information effectively through the use of structured vocabulary and a thoughtful approach to professional nursing practice. It begins to explore developing standards for paper or electronic documentation through collaboration with other stakeholders, including anesthesia care providers, surgeons, clinical directors, and informaticians. PMID- 11378947 TI - Perioperative care of the patient with Zenker's diverticulum. AB - There are a variety of surgical approaches to treat the symptomatic patient with a Zenker's diverticulum. Approaches to treat Zenker's diverticula include external excision (i.e., diverticulectomy) with or without myotomy; diverticulopexy, an endoscopic approach; or cricopharyngeal myotomy alone. This article discusses esophageal diverticula, treatment modalities, and the perioperative care of patients with Zenker's diverticula. PMID- 11378948 TI - Effects of warming therapy on pressure ulcers--a randomized trial. AB - Postoperative pressure ulcers are a common and expensive problem. Intraoperative hypothermia also is a common problem and may have a connection with impaired tissue viability. Researchers in this study hypothesized that intraoperative control of hypothermia may reduce the incidence of postoperative pressure ulcers. A randomized clinical trial (n = 338) was used to test the effects of using forced air warming therapy versus standard care. Results indicated an absolute risk reduction in pressure ulcers of 4.8% (i.e., 10.4% to 5.6%) with a relative risk reduction of 46% in patients who received warming therapy. Although not reaching statistical significance, the clinical significance of almost halving the pressure ulcer rate is important. A correlation between body temperature and postoperative pressure ulcers was established. PMID- 11378949 TI - Implementing a perioperative nursing fellowship program. AB - The population of nurses in the United States is aging. After suffering high OR nurse vacancy rates for several years, leaders at the Inova Health System Implemented a nurse fellowship program. This six-month didactic and precepted learning program is aimed at experienced RNs who do not have OR experience but would like to work in the OR. Two years after implementation, the OR nurse vacancy rate in the health system has decreased from 27% to 15.5%. PMID- 11378950 TI - Coping with the stress of surgery. PMID- 11378951 TI - Data fields for intraoperative records using the Perioperative Nursing Data Set. PMID- 11378952 TI - Reuse of single-use devices. Association of periOperative Registered Nurses. PMID- 11378953 TI - Hermeneutics as a methodology for textual analysis. PMID- 11378954 TI - The critical "nurse" in the circulating nurse role. PMID- 11378955 TI - Web alert. The chemistry of biological products. PMID- 11378956 TI - Therapeutic antibody production technologies: molecules, applications, expression and purification. AB - Antibody-based therapeutics are currently being tested in an increasingly diverse range of therapeutic modalities. Many different engineered formats for the antibody molecule and multiple methods for raising and tailoring binding specificities are currently available. Comparison of the relative function and efficacy of these molecules and the many competing methods for their production is crucial for making an informed selection of a new therapeutic entity. In addition, these choices may be influenced by the attached intellectual property burden. PMID- 11378957 TI - The development of genomics-derived pharmaceuticals. AB - Messenger RNAs for the large majority of human genes are now available. Most of those contained in the Human Genome Sciences database are capable of producing functional proteins. We have sequenced the clones of approximately 8000 that are believed to be involved in cell signaling, and have produced small amounts of the corresponding proteins. These are being screened for biological activity; four have been entered into clinical trials, with one demonstrating clinical activity thus far. PMID- 11378958 TI - An update on the state of the art of DNA vaccines. AB - DNA vaccines have been extensively studied in the past ten years and much is now known about their effectiveness and mode of action in animal models. Several DNA vaccines have been tested in phase I clinical trials, with mixed results. That is, DNA vaccines appear safe and well tolerated, but lack potency. This has led to the search for technologies that will enable sufficient potency for effectiveness in humans. PMID- 11378959 TI - In vitro protein display in drug discovery. AB - Nucleic acid-encoded libraries have been used at different stages of the drug discovery process for the identification of polypeptide ligands and for target identification. Traditionally, phage display screening systems have been used to explore large libraries of peptides and proteins. Lately, novel protein selection technologies have been developed that work entirely in vitro and use the polymerase chain reaction (PCR) rather than cells to amplify genetic material. The simplicity of the linkage between the protein and its encoding nucleic acid leads to several advantages, including the use of larger libraries without the biases of cell-based amplification, greater control over binding conditions and the ease with which PCR-based mutagenesis and recombination can be incorporated. This review focuses on the latest improvements in this new generation of in vitro protein display techniques and discusses their applications to the drug discovery process. PMID- 11378960 TI - Chemical protein synthesis methods in drug discovery. AB - The applicability of chemical protein synthesis over the last five years has been greatly expanded by significant increases in the size (up to 600 amino acids) and variety of accessible proteins, including new classes of proteins such as phosphoproteins, glycoproteins and integral membrane proteins. Chemical protein synthesis has also produced novel high-throughput screening and biosensor technology, and exciting lead compounds for the generation of powerful synthetic protein pharmaceuticals. PMID- 11378962 TI - Recent advances in the biosynthetic studies of lovastatin. AB - The fungal metabolite lovastatin and its derivatives are widely prescribed cholesterol-lowering drugs that act as potent inhibitors of (3S)-hydroxy-3 methylglutaryl-Coenzyme A reductase (HMG-CoA reductase). These drugs and a number of analogs that have been approved for use in humans are manufactured by fermentation in combination with subsequent chemical or microbial modification. This review highlights early work done in the elucidation of lovastatin biosynthesis involving the use of labeled precursors and the incubation of putative intermediates with cell-free extracts from various fungal sources. A series of more contemporary papers are also reviewed, describing the use of gene cloning to identify the various functions of the enzymes involved in the biosynthesis of lovastatin. In particular, overexpression, purification and the subsequent investigation of the various roles of lovastatin nonaketide synthase (LNKS) during lovastatin biosynthesis are discussed. PMID- 11378961 TI - Biosynthesis of hybrid peptide-polyketide natural products. AB - The structural and catalytic similarities between non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS) support the idea of combining individual NRPS and PKS modules for combinatorial biosynthesis. Recent advances in cloning and characterization of biosynthetic gene clusters for naturally occurring hybrid polyketide-peptide metabolites have provided direct evidence for the existence of hybrid NRPS-PKS systems, thus setting the stage to investigate the molecular basis for intermodular communication between NRPS and PKS modules. Reviewed in this article are biosynthetic data pertinent to hybrid peptide polyketide biosynthesis published up to late 2000. Hybrid peptide-polyketide natural products can be divided into two classes: (i) those whose biosyntheses do not involve functional interaction between NRPS and PKS modules; and (ii) those whose biosyntheses are catalyzed by hybrid NRPS-PKS systems involving direct interactions between NRPS and PKS modules. It is the latter systems that are most likely amenable to combinatorial biosynthesis. The same catalytic sites appear to be conserved in both hybrid NRPS-PKS and normal NRPS or PKS systems, with the exception of the ketoacyl synthase domains in hybrid NRPS-PKS systems which are unique. Specific linkers may play a critical role in communication, facilitating the transfer of the growing intermediates between the interacting NRPS and/or PKS modules. In addition, phosphopantetheinyl transferases with broad carrier protein specificity are essential for the production of functional hybrid NRPS-PKS megasynthetases. These findings should now be taken into consideration in engineered biosynthesis of hybrid peptide-polyketide natural products for drug discovery and development. PMID- 11378963 TI - SMR-type multidrug resistance pumps. AB - Multidrug resistance (MDR) efflux pumps in pathogenic microorganisms nullify the effects of antimicrobial drugs used in medicine. We have conducted phylogenetic analyses showing that these efflux pumps are associated with five superfamilies of transport systems. One of these, the drug/metabolite transporter (DMT) superfamily includes a family of small multidrug resistance (SMR)-conferring proteins that are discussed in detail in this review. A single microorganism such as Bacillus subtilis may possess multiple homologs of this family, and these homologs are believed to form both homo-oligomeric or hetero-oligomeric pumps, some of which export cationic drugs. The characteristics of some of these systems and the genes that encode them are described, with emphasis on the eight homologs encoded within the B subtilis genome. Anomalies and unanswered questions that provide impetus for future studies are presented. PMID- 11378964 TI - Stem cell collection filter system for human placental/umbilical cord blood processing. AB - BACKGROUND AND OBJECTIVES: The hydroxyethyl starch method and the Top & Bottom method have been used worldwide for the volume reduction of human placental/umbilical cord blood (PCB) units. To simplify the preparation of nucleated cell (NC) concentrates, we developed a new filter device--the stem cell collection filter system (SCF SYSTEM)--which can collect mononuclear cells (MNC) at a high recovery rate. MATERIALS AND METHODS: The SCF SYSTEM consisted of a filter and two bags. Multilayered polyethylene terephthalate non-wovens, coated with a hydrophilic polymer, were used as filter media. PCB units were filtered by gravity (n = 12). Red blood cells, platelets and plasma were drained into the drain bag, and the NC trapped on the filter media was collected in the recovery bag by reverse washing. Recovered cell fractions were evaluated. RESULTS: The volume of cell concentrate recovered was 27.4 +/- 2.2 ml (mean +/- SD, n = 12). The whole time required for processing was less than 30 min, and handling was very simple. The viability of recovered NC was 97.8 +/- 3.2%. The recovery of lymphocytes, monocytes and granulocytes was 79.5 +/- 16.9%, 79.8 +/- 20.4% and 39.0 +/- 19.5%, respectively. The recovery rate of granulocytes was significantly lower than that of monocytes and lymphocytes (P < or = 0.0001). The recovery rates of CD3+ cells, CD19+ cells and CD56+ cells were almost the same as that of MNC. The recovery rates of CD34+ cells, total colony-forming cells and long-term culture-initiating cells were 81.7 +/- 27.0% (n = 11), 80.8 +/- 27.7% (n = 12) and 75.0 +/- 18.4% (n = 2), respectively. CONCLUSION: The new filter system was shown to be efficient for PCB processing, encompassing a very simple handling procedure with a good recovery of haematopoietic progenitor cells. Hence, the SCF SYSTEM is potentially useful for the volume reduction of PCB units for cord blood banking. PMID- 11378965 TI - Evaluation of solutions for the storage of granulocyte colony-stimulating factor mobilized granulocyte concentrates. AB - BACKGROUND: High cell counts in granulocyte colony-stimulating factor (G-CSF) mobilized granulocytes are detrimental to concentrate storage. An eightfold dilution with autologous plasma improves storage, but this method is impractical. The purpose of this study was to identify an infusible solution that could be used in place of autologous plasma to dilute and store granulocytes. MATERIALS AND METHODS: Granulocytes collected from donors given dexamethasone (8 mg per os) and/or G-CSF (5 micrograms/kg subcutaneously [SQ]) were diluted eightfold in the following cell culture media: X-Vivo 10, Dulbecco's modified Eagle's minimal essential medium (DMEM) or Iscoves modified Dulbecco's medium (IMDM); or in the following infusible solutions: Plasma-Lyte A; Normosol R; lactated ringers, supplemented with 1% human serum albumin and 50 mM histidine (LRAH); or Plasma Lyte A supplemented with 50 mM histidine buffer or 25 mM HEPES buffer plus 1% human serum albumin. The granulocytes were stored for 48 h at room temperature. White blood cell (WBC) counts, WBC viability and pH were measured after approximately 2 h, 24 h and 48 h of storage. RESULTS: Cell counts, viability and pH were maintained after 2 h, 24 h and 48 h in cells stored in the three cell culture media. The pH fell slightly after 48 h to 6.86 +/- 0.10 in granulocyte concentrates diluted in LRAH, but fell to a greater extent after 24 h and 48 h, to 6.36 +/- 0.23 (48-h value) in granulocyte concentrates diluted in Plasma-Lyte A and to 6.40 +/- 0.19 (48-h value) in granulocyte concentrates diluted in Normosol R. The cell counts of concentrates diluted in LRAH were stable for 48 h, but fell in granulocyte concentrates stored in Plasma-Lyte A and Normosol R. Plasma-Lyte A supplemented with histidine maintained the pH of diluted granulocyte concentrates better than Plasma-Lyte A supplemented with HEPES; 6.91 +/- 0.10 and 6.65 +/- 0.11, respectively, after 24 h. Cell counts were maintained best in granulocyte concentrates diluted in Plasma-Lyte A supplemented with albumin and one or both of the buffers. CONCLUSIONS: Culture media were best for granulocyte storage, but they are not approved for in vivo use. Infusible solutions are not buffered adequately and lack sufficient protein, but infusible solutions, such as lactated Ringer's solution or Plasma-Lyte A supplemented with buffers and albumin, hold promise as effective and licensable solutions for granulocyte storage. PMID- 11378966 TI - Genotyping fetal DNA by non-invasive means: extraction from maternal plasma. AB - BACKGROUND AND OBJECTIVES: Identification of fetal DNA in maternal plasma may allow genetic analysis without the use of invasive techniques. The aim of this study was to extract DNA from maternal plasma, identify fetal material through the presence of SRY or RHD gene sequences and assess the reliability of these results. MATERIALS AND METHODS: A polymerase chain reaction (PCR) method of a commercial kit was used with primers for SRY or exon 10 of the RHD gene sequence. RESULTS: Multiple plasma samples were collected from 60 women who were evaluable for either SRY or RHD, or both, fetally derived DNA sequences. Fetal DNA was present in the plasma throughout the pregnancies and for some hours or days after delivery. CONCLUSION: Fetal DNA can be reliably detected in maternal plasma from early in pregnancy and normally is cleared within days of delivery. PMID- 11378967 TI - Time-dependent increase of aluminium in human albumin solutions: the role of initial citrate content. PMID- 11378968 TI - Red cell transfusions in neonatal care. PMID- 11378969 TI - Tumour immunotherapy: new tools, new treatment modalities and new T-cell antigens. AB - Tumour immunology has seen many exciting developments in the last few years. In addition to tumour antigens that are defined by antitumour T- and B-cell responses in patients, the human telomerase reverse transcriptase has been identified by 'reverse immunology' as the first truly universal tumour antigen. Molecular remission has been associated with a cancer vaccine that targets the clonal idiotype of B-cell malignancies, and sophisticated cellular vaccines (including fusions of tumour cells and antigen-presenting cells) have demonstrated promising results. Moreover, our capabilities of measuring immunity have been significantly enhanced by novel technology, such as major histocompatibility complex (MHC)-peptide tetramers and ELISPOT analysis. We are now capable of tracking antigen-specific T cells at a single cell level. This review will analyse recent developments and highlight some important issues that need to be addressed in the future. PMID- 11378970 TI - Blood donors infected with the hepatitis B virus but persistently lacking antibodies to the hepatitis B core antigen. AB - BACKGROUND AND OBJECTIVES: Antibodies to the core of hepatitis B virus (anti-HBc) are considered to be the best serologically reliable markers of hepatitis B virus (HBV) infection. Through a national epidemiological survey, two young and first time blood donors, originating from HBV-endemic areas, were identified as HBV carriers with an absence of anti-HBc reactivity. MATERIALS AND METHODS: We followed up these two subjects in order to investigate the evolution of their HBV serological profiles. Nucleotide sequencing was performed of the entire pre-C/C region of the strains infecting these donors. RESULTS: The same serological profile of active viral replication with an apparent persistent lack of anti-HBc and normal alanine aminotransferase (ALT) levels was found for both subjects throughout a follow-up of 19 months and 4 months, respectively. Neither donor was immunocompromised. Nucleotide sequence analysis of the pre-C/C region did not show mutations or deletions in encoded proteins. CONCLUSION: The hypothesis of an in utero HBV infection responsible for an immune tolerance to HBV seems to be the most probable explanation for this particular immunological situation. Such occurrences in the blood donor population are probably rare as less than 0.1% of hepatitis B surface antigen (HBsAg)-positive donors exhibit such a profile, in our experience. Moreover, this phenomenon does not impose a risk of HBV transmission by blood donation, as the exclusion of HBV-infected blood donation is based on HBsAg detection. However, such a risk might be encountered with the hepatitis C virus (HCV) for which at present only antibodies to HCV are screened. PMID- 11378971 TI - Preoperative autologous blood donation by elderly patients undergoing orthopaedic surgery. AB - BACKGROUND AND OBJECTIVES: To assess the feasibility of a programme of predeposit in elderly patients undergoing elective orthopaedic surgery. PATIENTS AND METHODS: We retrospectively studied 789 elderly patient candidates (> 65 years of age) for orthopaedic surgery (total hip and knee replacement and spinal surgery), who were undergoing a programme of preoperative autologous blood donation (PABD) in our city hospital between January 1990 and December 1998. RESULTS: Six hundred and eighty-eight patients (87.2%) were transfused with autologous blood; 128 (16.2%) also received allogeneic blood. Hip arthroplasty revision was characterized by the greatest blood consumption. The predeposit programme was discontinued in 96 patients (12.2%) because of the following complications: the onset of anaemia (11.0%); vasovagal reactions (0.5%); lack of venous access (0.4%); or cardiac complications (0.2%). No episodes of reaction to autologous transfusion were recorded. CONCLUSIONS: Our study confirms the feasibility of PABD in elderly patients undergoing elective orthopaedic surgery. PMID- 11378972 TI - Quantitative study designs used in quality improvement and assessment. AB - This article describes common quantitative design techniques that can be used to collect and analyze quality data. An understanding of the differences between these design techniques can help healthcare quality professionals make the most efficient use of their time, energies, and resources. To evaluate the advantages and disadvantages of these various study designs, it is necessary to assess factors that threaten the degree with which quality professionals may infer a cause-and-effect relationship from the data collected. Processes, the conduits of organizational function, often can be assessed by methods that do not take into account confounding and compromising circumstances that affect the outcomes of their analyses. An assumption that the implementation of process improvements may cause real change is incomplete without a consideration of other factors that might also have caused the same result. It is only through the identification, assessment, and exclusion of these alternative factors that administrators and healthcare quality professionals can assess the degree to which true process improvement or compliance has occurred. This article describes the advantages and disadvantages of common quantitative design techniques and reviews the corresponding threats to the interpretability of data obtained from their use. PMID- 11378974 TI - Continuous improvement evaluation: a framework for multisite evaluation studies. AB - This article proposes the use of the continuous improvement evaluation (CIE), a framework for multisite demonstration or evaluation studies. This framework is designed for studying intervention programs that change during the evaluation. The development of family drug courts is provided as an example. CIE relies on outcome data collected over time and benchmarked across similar cases in comparison sites; thus, this study was designed to collect data on effectiveness of intervention programs at multiple sites and over time. A weight is calculated for similarity of any two cases based on features they share. In statistical process control charts, these weights are used to compare outcomes at the site against the average of similar cases in comparison groups. Once data are benchmarked, program staff meet to discuss process changes that have led to improvements in outcomes. To ensure that intervention programs have access to evaluation reports on demand, information technology is used to collect, clean, and pool data. Computers generate study reports, and evaluators review reports after release to clients. Statistical tools can be used to evaluate changing programs. Traditional evaluators may be concerned about some threats to validity associated with CIE. The article concludes with a discussion of typical threats to validity and how these threats are addressed in the CIE framework. PMID- 11378973 TI - Margaret E. O'Kane on healthcare accreditation. Interview by Wanda Bishop and Kevin C. Park. AB - Since 1990, Margaret E. O'Kane has served as president of the National Committee for Quality Assurance (NCQA), an independent, not-for-profit organization whose mission is to improve the quality of healthcare everywhere. Under O'Kane's leadership, NCQA has developed broad support among the employer and health plan communities; today many Fortune 100 companies will do business only with NCQA accredited health plans. About three quarters of the nation's largest employers use Health Plan Employer Data and Information Set (HEDIS) data to evaluate the plans that serve their employees. O'Kane was named Health Person of the Year in 1996 by the journal Medicine & Health. She also received a 1997 Founder's Award from the American College of Medical Quality, recognizing NCQA's efforts to improve managed care quality. In 1999, O'Kane was elected a member of the Institute of Medicine. In 2000, she received the Centers for Disease Control and Prevention (CDC) Champion of Prevention award, the agency's highest honor. CDC names a Champion of Prevention infrequently and only when an individual has made a truly notable contribution to advancing preventive healthcare. O'Kane began her career in healthcare as a respiratory therapist and has a master's degree in health administration and planning from Johns Hopkins University. PMID- 11378975 TI - Development of a computerized screening system to identify substance abuse in primary care. AB - The Drug and Alcohol Problem Assessment for Primary Care (DAPA-PC), developed under a contract from the National Institute on Drug Abuse, is a comprehensive screening system developed for quickly identifying and addressing substance abuse and related problems in a primary care setting. The DAPA-PC system includes a two level screening instrument, resources for scoring and reporting results, motivational messages, treatment referrals, and informative links, which together address the multifaceted needs of patients dealing with addiction problems and their providers. The need for such a system, preliminary reactions to the instrument, and future course of research are discussed. PMID- 11378976 TI - Gender differences in utilization of exercise treadmill testing: a claims-based analysis. AB - Significant data exist to suggest that women with coronary artery disease (CAD) have worse outcomes than men. One explanation is that women present with more advanced disease because of a lack of screening and early detection. To examine this, we performed a claims-based analysis of exercise treadmill testing (ETT) within an urban managed care population. We analyzed all claims for 57,793 covered lives. Utilization rates for ETT were calculated for men and women by age group. ETT utilization between men and women demonstrated significant differences: 49.2 tests/1,000 men versus 25.0 tests/1,000 women (p < 0.001). This 2:1 ratio was observed across multiple age strata. Significant differences in ETT utilization exist between men and women, even in older populations in which the incidence of CAD in women exceeds that in men. ETT often is the initial screening test for detecting CAD; these gender differences in ETT utilization may result in underdetection of CAD in women and treatment at later stages of disease when interventions are less effective. PMID- 11378977 TI - Factors associated with Medicare beneficiary complaints about quality of care. AB - This article examines the number and types of formal complaints about quality of care that were made by Medicare beneficiaries and submitted to the California Peer Review Organization (PRO) during the period July 1, 1995-December 30, 1996. Logistic regression models were used to analyze the complaints in terms of sociodemographic factors, enabling factors (income and health maintenance organization [HMO] membership), diagnoses, and primary service providers. The complaint rate was found to be very low, and only 13% of complaints were confirmed by the PRO. HMO members and members receiving physician care and outpatient/emergency room care were more likely to complain about denials of or delays in services or the failure to be referred to specialists than were members in fee-for-service plans and those receiving other types of provider care. Complaints about poor nursing care were associated with receiving skilled nursing/rehabilitation care. Complaints about care that resulted in injury were associated with the denial of care, failure to be referred to a specialist, poor medical care, and poor communications. Complaints about care that led to disability were associated with medical errors, whereas those that led to death were associated with misdiagnosis and premature hospital discharge. It would be valuable for PROs to focus their complaint review efforts on common types of complaints in different settings. A review of PRO procedures should be undertaken to understand why so few complaints are submitted and confirmed. PMID- 11378978 TI - Application of a hybrid analysis method for a Web-enabled information system: a case study. AB - A successful developmental effort frequently is defined as one that produces a deliverable that is satisfactory to the client, on time, within budget, and has had no adverse impact on ongoing business operations. This article describes through a case study approach the activities that one project used to ensure a successful development effort. The author advocates a hybrid methodological approach to systems analysis and design that is tailored for the specific project. The article also describes the project and people management practices used to support the developmental effort. PMID- 11378979 TI - Systems analysis and design methodologies: practicalities and use in today's information systems development efforts. AB - Historically, systems analysis and design methodologies have been used as a guide in software development. Such methods provide structure to software engineers in their efforts to create quality solutions in the real world of information systems. This article looks at the elements that constitute a systems analysis methodology and examines the historical development of systems analysis in software development. It concludes with observations on the strengths and weaknesses of four methodologies and the state of the art of practice today. PMID- 11378980 TI - Challenges in incorporating a quality strategy into information systems development efforts. AB - Quality is an essential element of conducting successful information systems (IS) initiatives. While quality is a critical issue, statistics indicate that between one and two thirds of IS projects fail and of the most expensive software projects, about half eventually will be canceled for being out of control. This article examines the three phases of quality management and how these relate to IS development. This article illustrates the challenges faced in incorporating quality into an IS development effort and concludes with observations about the general states of IS development quality. PMID- 11378981 TI - BPR (business process redesign) and systems analysis and design: making the case for integration. AB - The question of what role business process redesign (BPR) should play in information systems development is one that draws strong opinions on either side of the issue. Some argue that BPR is absolutely necessary in forging a synergy between organizational structure, people, technology, and tasks. However, consultation and application development continue today without sufficient workflow analysis, as if BPR was irrelevant altogether. This article examines major BPR theories and the similarities and differences between BPR and information systems development and engages in a discussion of the merits of including BPR in any information systems development project. PMID- 11378982 TI - CASE tools and UML: state of the ART. AB - With increasing need for automated tools to assist complex systems development, software design methods are becoming popular. This article analyzes the state of art in computer-aided software engineering (CASE) tools and unified modeling language (UML), focusing on their evolution, merits, and industry usage. It identifies managerial issues for the tools' adoption and recommends an action plan to select and implement them. While CASE and UML offer inherent advantages like cheaper, shorter, and efficient development cycles, they suffer from poor user satisfaction. The critical success factors for their implementation include, among others, management and staff commitment, proper corporate infrastructure, and user training. PMID- 11378984 TI - Web-based e-mail communications: issues in responsive service. AB - Women's College Hospital (WCH), Toronto, Canada, embarked on a review of 5 years of Web-based e-mail communications to address broad issues of managerial policy and procedure as related to the institution's expanding role in Web-based communication. The hospital looked at who was sending messages, why they were communicating, and how the institution should respond to the new demands placed on them by virtual communicators. The article provides a brief discussion of the results of the review and points out that attention must be given to a broad range of organizational challenges and opportunities posed by such communications. Suggestions related to how health care organizations may respond better to and serve their virtual audiences are offered. PMID- 11378983 TI - The Payment Error Prevention Program (PEPP): reducing Medicare payment errors in prospective payment system hospitals. AB - Peer Review Organizations (PROs) are charged by the Health Care Financing Administration (HCFA) to assist in protecting the integrity and solvency of the Medicare program. Recent audits of the Medicare program from the Office of Inspector General (OIG) revealed that more than $12 billion Medicare dollars in 1998 were spent in improper payments, with more than 25% attributed to prospective payment system (PPS) hospitals. The Payment Error Prevention Program (PEPP) is an initiative designed by HCFA in 1999 to assist PROs in meeting the goal of reducing payment errors in PPS hospitals. PEPP is implemented through the development of quality improvement (QI) methodologies. These projects are designed to achieve measurable improvements in processes and outcomes of payment errors. PEPP works to reduce payment errors at PPS hospitals through cooperative efforts with Ohio agencies and licensing boards, federal law enforcement organizations, HCFA contractors, hospital medical staffs, and medical and osteopathic associations. PMID- 11378985 TI - The ability to successful implement information technology is a challenge irrespective of the environment in which it is used (e.g., health care, banking, manufacturing). PMID- 11378986 TI - [Clinical training and individual motivation]. PMID- 11378987 TI - [From the past--to enrich the future]. PMID- 11378988 TI - [Sociology and practical training of nursing students]. PMID- 11378989 TI - [The flavour of nursing]. PMID- 11378990 TI - [First and hand tied to the cigarette]. PMID- 11378991 TI - ["Listen, family!", a center for appeal to UNAFAM (National Union of Friends and Family of Patients)]. PMID- 11378992 TI - [A specific case: evaluate for treatment]. PMID- 11378994 TI - [The 85th Congress of the Japanese Society of Legal Medicine. Kurume, Japan. April 11-13, 2001. Abstracts]. PMID- 11378993 TI - [From the essential to the complexity]. PMID- 11378995 TI - Grandmothers raising grandchildren: are they at increased risk of health problems? AB - Many grandparents faced with the need to raise their grandchildren have health risks that could destabilize any family structure established for the child. This is especially problematic for grandparents of color who have higher health risk factors that their white counterparts. This article describes the physical and mental health status and behaviors of 100 African American grandmothers who are the primary caretakers for their grandchildren. The findings suggest the physical functioning of the grandmothers is at a level that could jeopardize the quality of life with their grandchildren. In spite of their diminished physical capacity, the grandmothers reported that their emotional state is equal to or better than the general population. Suggestions for community practice are provided. PMID- 11378996 TI - Medicare and prescription drugs: prospects for reform. PMID- 11378997 TI - Pregnancy: a time to break the cycle of family violence. PMID- 11378998 TI - Cognitive-behavioral therapy with a six-year-old boy with separation anxiety disorder: a case study. PMID- 11378999 TI - Women's health needs special treatment. PMID- 11379000 TI - Social support: a critical factor in women's health and health promotion. AB - Social networks and social support have been found to be beneficial to the health of individuals in a variety of ways--reducing mortality rates, improving recovery from serious illness, and increasing use of preventive health practices. Social relationships appear to be particularly important to women. Suggestions for health promotion and disease prevention activities include the use of buddy systems, feminist pedagogical techniques, group activities, and linkage to natural supports. Health promotion with women is an underdeveloped area of social work practice with great potential to increase the health of client populations. PMID- 11379001 TI - A female perspective on living with HIV disease. AB - The study discussed in this article explored women's views of the positive and negative aspects of life with HIV. Even in the face of a stigmatizing physical illness and with elevated levels of depression and anxiety, the 55 women interviewed for the study were able to identify a large number of positive events; for many, HIV served as a motivating force for positive change. Common negative experiences included physical symptoms, a limited life span, alienation, and stigma. Results suggest that whereas women demonstrate a remarkable capacity to adapt, there are a number of specific areas where social services and community interventions can be targeted. PMID- 11379002 TI - Social work intervention and failure to thrive in infants and children. AB - Failure to thrive (FTT) is a diagnostic term used to describe infants and children who fail to grow and develop at a normal rate. There has been limited literature focused on the role social workers play in working with children who fail to thrive. Because the focus of the current health care system is on using an interdisciplinary team approach, it is crucial that social workers make a place for themselves on treatment teams and continue to use their skills to provide comprehensive treatment. This article educates social workers about FTT and addresses the roles social workers have in the treatment of children who fail to thrive and how their services provide a therapeutic and team-oriented approach. PMID- 11379003 TI - The decision-making process for requests for late termination of pregnancy in Israel. AB - This article outlines and reflects on the current procedure in Israel for requesting late pregnancy terminations and the subsequent decision-making processes. The study population consisted of 183 women who requested late termination of pregnancy (LTP) in Israel during the years 1995-97. The main causes for requests were fetal anomalies and late application. Eighty-two percent of requests were approved, and approval could be predicted by the week of gestation and the reason for application. Ethical dilemmas are described. The need to enhance professional support for women who request LTP, whether they receive approval or not, should be addressed by the health system and social workers in health care. PMID- 11379004 TI - [59th annual meeting of the Japanese Society of Public Health. 2000. Proceedings]. PMID- 11379005 TI - [30th annual meeting of the Japenese Society of Neuropsychopharmacology. Sendai, Japan. October 26-27, 2000. Abstracts]. PMID- 11379006 TI - Effects of estrous cycle and steroid replacement on the expression of leptin and uncoupling proteins in adipose tissue in the rat. AB - Among other actions, leptin has been suggested to increase energy expenditure and to modulate the menstrual cycle. In fact, the main effect of leptin seems to be modulating the sympathetic nervous system and gonadotropin-releasing hormone pulsatility. We investigated whether changes in the plasma steroid concentrations during the estrous cycle and after ovariectomy and steroid replacement can modulate plasma leptin levels, adipose tissue leptin mRNA expression, and some of the candidates for mediating energy expenditure (uncoupling proteins (UCP) 1, 2, and 3 mRNA) in white and brown adipose tissue. Rats in estrous cycle or ovariectomized rats with or without estradiol or progesterone replacement therapy for 18 days were studied. Plasma leptin, insulin, estradiol and progesterone were measured with radioimmunoassays. Leptin mRNA expression was measured in subcutaneous, periovarian and mesenteric white adipose tissue and in interscapular brown adipose tissue. Expression of UCP 1, 2, and 3 mRNA in periovarian white and brown adipose tissue was analyzed. Plasma leptin levels were significantly decreased in the estrous (1.1 +/- 0.4 ng/ml) compared with the pro-estrous (1.7 +/- 0.4 ng/ml, F = 3.0, p = 0.046) phase of cycle. UCP1 mRNA levels in brown adipose tissue were more elevated during pro-estrus than during metestrus (F = 3.17, p = 0.039). Gene expressions of leptin, UCP2 or UCP3 mRNA did not change significantly during the cycle. In ovariectomized rats, estradiol and/or progesterone treatment had no effect on plasma leptin levels. Gene expression analysis of leptin and UCP1, 2 and 3 in adipose tissue was not affected by steroid replacement. In conclusion, the estrous cycle appears to have a minor effect on modulation of leptin and uncoupling proteins. Only plasma leptin levels and expression of UCP1 mRNA are modestly elevated during the estrous cycle in the rat. Since estrogen and/or progesterone substitution in ovariectomized rats does not affect circulating leptin concentration or expression of leptin and UCPs in adipose tissue, it is unlikely that steroids play a major role in their regulation. PMID- 11379007 TI - Expression of the gap junction connexins Cx43, Cx45 and Cx26 in human uterine leiomyomata. AB - Uterine leiomyomata of 34 premenopausal women undergoing leiomyomectomy or hysterectomy, and in four cases the corresponding myometrium, were collected at laparotomy or laparoscopy to investigate the ability of these benign smooth muscle cell tumors to express different connexins. Immunohistochemical and Northern blot analyses were performed for the characterization of the expression of connexins Cx43, Cx45, Cx26 and Cx32. Immunofluorescence revealed the presence of Cx43 in most leiomyomata. Only seven leiomyomata lacked Cx43 expression. Cx45 was expressed in 13, a weak Cx26 immunostaining was found in seven cases, whereas Cx32 could not be detected. No correlations between the 17 beta-estradiol or progesterone serum levels and the expression patterns of the connexins Cx43, Cx45 and Cx26 could be observed. Gonadotropin-releasing hormone (GnRH)-agonist or progestin treatment did not influence the connexin expression pattern. Northern blot analyses confirmed these results; however, transcripts of Cx26 were not detectable. Connexin transcripts between myomata and the corresponding myometrium showed no obvious differences. Our data show that uterine leiomyomata are capable of expressing different connexins comparable to the corresponding myometrium, but do not respond to different hormonal conditions. The ability to express the appropriate connexins could explain why these tumors, though developing independently of hormonal levels, are still differentiated benign smooth muscle tumors. PMID- 11379008 TI - Investigations on the genetic polymorphism in the region of CYP17 gene encoding 5'-UTR in patients with polycystic ovarian syndrome. AB - Polycystic ovarian syndrome (PCOS) is a prevalent endocrine disorder, which is the most common cause of anovulatory infertility and hirsutism. It is caused by an overproduction of androgens in theca cells. In the ovary, androgen synthesis is regulated by 17 alpha-hydroxylase/17,20-lyase enzyme complex containing P450c17 (CYP17). In some individuals, the promoter region of CYP17 gene contains a T-->C substitution that creates an Sp1 site at position-34. This polymorphism generates a recognition site for the MspA1 restriction enzyme. The objective of the study was to investigate the frequency of T-->C substitution of CYP17 gene promoter in women with PCOS and elucidate its role in the pathogenesis of the syndrome. Another aim of the study was to compare the results with the levels of the hormones luteinizing hormone, follicle-stimulating hormone, testosterone and estradiol. The present investigation involved a group of 55 women with PCOS and 56 healthy women without symptoms of PCOS. Following digestion with restriction enzyme MspA1, it was demonstrated that the polymorphic A2 allele is no more frequent in women with PCOS than in healthy women. In the PCOS group, the hormonal profiles were not significantly different in the carriers of a normal allele A1A1 from the carriers of A1A2 and A2A2 alleles. It is concluded that T- >C polymorphism of CYP17 gene is not associated with steroid hormone synthesis in PCOS and is not the primary genetic defect in this disease. PMID- 11379009 TI - Fertility outcome: long-term results after laparoscopic myomectomy. AB - From 1991 to 1998, 29 patients desiring a pregnancy underwent laparoscopic myomectomy for symptomatic myomas measuring 5.4 +/- 3.6 cm (mean +/- SD) (median 5; range 1-9). The overall rate of intrauterine pregnancy was 65.5% (19 pregnancies; two patients had two pregnancies each). Results were analyzed in relation to different preoperative clinical conditions. Out of nine patients with other infertility factors associated with uterine myomas, three (33.3%) became pregnant; out of 10 infertile patients with no other associated infertility factors, seven (70%) became pregnant; out of 10 patients to whom myomectomy was performed for the rapid growth of the tumor or for myoma encroaching on the cavity, nine (90%) had a pregnancy. Nine patients (73.4%) had a Cesarean section (one twice), four (26.6%) had spontaneous vaginal delivery, one patient had a serious placental failure at the 28th week, and four patients (19%) miscarried. Two pregnancies are still in progress (one in a patient with previous miscarriage). Out of 21 pregnancies, the viable term delivery rate was 57.14%. No uterine ruptures were observed. The pregnancy rate after laparoscopic myomectomy was similar to that reported in other studies after laparotomic myomectomy. It is concluded that laparoscopic myomectomy is a reliable procedure even in the presence of multiple or enlarged myomas. Moreover, our pregnancy rate and pregnancy outcome seem to indicate that both desire for pregnancy and infertility prior to surgery should not be exclusion criteria for the laparoscopic approach. PMID- 11379011 TI - Polycystic ovary syndrome: relationship between insulin sensitivity, sex hormone levels and ovarian stromal blood flow. AB - Polycystic ovary syndrome (PCOS) is the most common cause of menstrual disorders, and is characterized by chronic anovulation, hyperandrogenism and infertility. In recent years, it has become apparent that PCOS is also associated with hyperinsulinemia that is probably central to the pathogenesis of PCOS. As a peculiar vascular pattern has been reported to be present in PCOS, the aim of this study was to investigate intraovarian stromal vascularization in PCOS patients and its possible correlation with sex hormones, gonadotropins and insulin levels. Twenty-eight oligomenorrheic or amenorrheic patients with PCOS and 14 eumenorrheic women with a PCOS-like ovarian pattern undergoing endocrine screening and ultrasound color Doppler intraovarian blood flow were recruited to the study. Ten healthy women with regular menses represented the control group. Hormonal assays (follicle-stimulating hormone (FSH), luteinizing hormone (LH), androstenedione, testosterone, sex hormone-binding globulin (SHBG) and estradiol), oral glucose tolerance test (OGTT), baseline and glucose-induced insulin levels, and transvaginal ultrasonographic and color Doppler analysis (pulsatility index (PI), resistance index (RI) and velocity (Vmax) of ovarian stromal flow) were performed in all participants in the early proliferative phase. Endocrine values showed significant differences in PCOS patients compared with PCOS-like women and controls, while PI and RI indices were significantly higher in controls. PCOS patients were divided into hyperinsulinemic (n = 16) and normoinsulinemic (n = 12). Androstenedione was significantly higher (p < 0.01) in the hyperinsulinemic than in the normoinsulinemic patients and controls, while SHBG was significantly (p < 0.01) lower in the hyperinsulinemic group. Analysis of color Doppler intraovarian vascularization showed a significantly lower RI and a higher Vmax in the hyperinsulinemic subjects than in the normoinsulinemic PCOS patients and controls. An increased stromal blood flow was observed in the PCOS and PCOS-like patients by transvaginal color Doppler evaluation, but this technique is not able to differentiate these two similar ovarian patterns. However, hyperinsulinemic PCOS patients had an increased vascularity of the ovarian stroma. A strong correlation between hyperinsulinemia, hyperandrogenism and low SHBG levels was evidenced, and a hyperinsulinemia-induced mechanism for ovarian stromal angiogenesis is discussed. PMID- 11379010 TI - Synthetic estrogens-mediated activation of JNK intracellular signaling molecule. AB - Signal transduction pathways regulate the transmission of specific signals to the cells from the surface to the nucleus. Activation of protein kinases such as JNKs (c-jun amino-terminal kinase), a subgroup of the mitogen activated protein kinase (MAPK) family, results in regulation of important cellular functions like cell growth and differentiation. The involvement of estrogens in stimulation of growth of already transformed breast cancer cells in vivo and in vitro accompanied with activation of JNKs prompted us to investigate the role of synthetic estrogens in the regulation of JNK expression. T 47D breast cancer cells were incubated with the synthetic estrogens, ethinylestradiol (10(-9)M) and 17 beta-estradiol valerate (10(-9)M), epidermal growth factor (EGF) (10 ng/ml) and the natural estrogen, 17 beta-estradiol (10(-9)M), for 5 minutes. The same experiments were repeated after pretreatment of the cells with ICI 182780 for 24 hours. EGF as well as natural and synthetic estrogens stimulated proliferation. This effect was reversed by the estrogen receptor blocker ICI 182780, but only in the case of both natural and synthetic estrogen. Like 17 beta-estradiol, synthetic estrogens induced a rapid and transient activation of JNK kinase. ICI 182780 blocked this effect, but not that mediated by EGF. Ethinylestradiol used in oral contraceptives, and 17 beta-estradiol and 17 beta-estradiol valerate for hormone replacement therapy, are able to activate JNK. The estrogen receptor is necessary for JNK activation upon estrogen stimulation. PMID- 11379012 TI - Oral estrogen replacement therapy increases forearm reactive hyperemia accompanied by increases in serum levels of nitric oxide in postmenopausal women. AB - The present study sought to determine the correlation between the vasodilator response of forearm resistance vessels and the circulating levels of nitric oxide (NO) after the administration of oral estrogen for 12 weeks to postmenopausal women. We classified postmenopausal women into two groups: those treated with conjugated equine estrogen (0.625 mg daily) orally for 12 weeks (n = 24) or those who received no treatment (control group, n = 8). Forearm blood flow was measured using strain-gauge plethysmography during hyperemia to evaluate endothelium dependent vasodilation, and after sublingual nitroglycerin administration to evaluate endothelium-independent vasodilation. Serum levels of nitrite/nitrate (metabolites of NO) and lipid parameters were measured. Basal forearm blood flow, body weight and heart rate were similar in each group. After 12 weeks of estrogen administration, the maximal forearm blood flow response during reactive hyperemia and the serum level of nitrite/nitrate each showed a significant increase from 26.9 +/- 1.9 to 37.9 +/- 3.5 ml/min per 100 ml tissue (p < 0.01), and from 25.2 +/- 2.2 to 37.5 +/- 3.7 mumol/l (p < 0.05), respectively. No increases were seen in controls. The changes in forearm blood flow after sublingual nitroglycerin were similar before and after 12 weeks of estrogen administration. The increase in maximal forearm blood flow with reactive hyperemia was significantly correlated with the increase in nitrite/nitrate in the group administered estrogen (r = 0.48, p < 0.05). There was no significant correlation between maximal forearm blood flow with reactive hyperemia, nor any change in serum lipids, blood pressure or other parameters. In conclusion, the 12-week administration of oral estrogen increased forearm reactive hyperemia in postmenopausal women, probably via an increase in the production of NO. PMID- 11379013 TI - Long-term effect of bilateral ovariectomy on endothelial function in aortic rings of spontaneously hypertensive rats: role of nitric oxide. AB - Endothelial dysfunction associated with both menopause and hypertension could be one of the possible explanations for increased cardiovascular morbidity and mortality in hypertensive postmenopausal women. The aim of the present study was to investigate the long-term effect of menopause (bilateral ovariectomy) on endothelial function in isolated aortic rings of spontaneously hypertensive rats (SHR). Aortic rings were suspended in organ chambers filled with physiological salt solution (95% O2, 5% CO2, 37 degrees C), and isometric tension was measured. In studies designed to assess the tone-related release of nitric oxide (NO) from phenylephrine-precontracted aortic rings, we found that vasoconstriction induced by L-NAME was greater in aortic rings from sham-ovariectomized SHR (SHAM SHR) than in those obtained from ovariectomized SHR (OVX SHR). Concentration-related relaxant responses to superoxide dismutase were significantly greater in the SHAM SHR than in the OVX SHR. In contrast, receptor-mediated release of NO was not altered by ovariectomy, as deduced from acetylcholine (ACh) concentration responses curves. Responses to the exogenous NO donor sodium nitroprusside (SNP) were also identical in both ovariectomized and sham-ovariectomized groups, ruling out differences in smooth muscle reactivity to NO. These results show that NO release is impaired in OVX SHR, an animal model of simultaneous hypertension and menopause. PMID- 11379014 TI - Adrenarche, pubertal development, age at menarche and final height of full-term, born small for gestational age (SGA) girls. AB - Children born small for gestational age (SGA) may present advanced bone maturation in childhood and reduced final height. The objectives of the study were to evaluate adrenarche, pubertal development, age at menarche and final height in full-term born-SGA girls. Twenty-four girls (12 born-SGA and 12 matched controls) were evaluated at 6-7.5 years of age for clinical signs of puberty and dehydroepiandrosterone sulfate (DHEAS) levels, as a marker of adrenarche. Thirty eight girls (19 born-SGA and 19 matched controls) were evaluated at 17.5-18.5 years of age to assess final height, sexual maturation and age at menarche. SGA girls had a mean final height (160.1 cm vs 165.8 cm, p < 0.01) and mean weight (52.1 kg vs 56.5 kg, p < 0.05) significantly lower than controls. Controls had a mean final height significantly higher than their mean target height. Sexual maturation was at stage 5 of Tanner's staging in SGA girls and control subjects. SGA girls had a slightly anticipated puberty (9.9 vs 10.4 years for initial breast development) and a lower age at menarche (11.9 vs 12.3 years). At 6-7.5 years of age, SGA females and controls did not show any difference for clinical signs of puberty; however, DHEAS levels (0.75 + 0.18 microgram/ml vs 0.57 + 0.22 microgram/ml, p < 0.05) were significantly higher in SGA girls than in control subjects. We concluded that full-term born-SGA females have impaired final height and weight in adolescence but substantially normal sexual maturation and age at menarche. Increased DHEAS levels before puberty in born-SGA girls may predispose to increased bone maturation in childhood with a reduced final height. In our population a progressive increment in final stature is evident. PMID- 11379015 TI - Cycle and age-related changes in corticotropin-releasing hormone levels in human endometrium and ovaries. AB - Corticotropin-releasing hormone (CRH) is synthesized in most female reproductive tissues such as the ovaries and the uterus. In the non-pregnant uterus, it is mainly produced by epithelial cells of the endometrium. Recent in vitro experimental findings show that endometrial CRH is under the positive control of progesterone, participating in the decidualization process of endometrial stroma and the progression of blastocyst implantation. CRH is also produced in the thecal compartment of the human ovary, controlling ovarian steroid hormone biosynthesis. In the present study we compared the concentration of immunoreactive CRH (ir-CRH) in biopsies from proliferative and secretory human endometria, and from pre- and postmenopausal human ovaries. We found that the concentration of ir-CRH was significantly higher in the secretory (92 +/- 8 pg/mg protein; n = 10) than the proliferative (75 +/- 9 pg/mg protein; n = 12; p < 0.05) endometria. This observation supports the experimental in vitro findings associating endometrial CRH in intrauterine phenomena of the secretory phase of the menstrual cycle (decidualization and implantation). Additionally, we have shown that the concentration of ir-CRH was significantly higher in the premenopausal (125 +/- 12 pg/mg protein; n = 14) than the postmenopausal (100 +/- 12 pg/mg protein; n = 12; p < 0.05) ovaries, suggesting that ovarian CRH is related to normal ovarian function during the reproductive lifespan. PMID- 11379016 TI - [Characterization of Ru(bpy)(3)2+ and Ru(phen)(3)2+ complexes with silver(i) ion of ground state in propylene-carbonate]. AB - Phosphorescent bimolecular and termolecular exciplexes of 3Ru(bpy)(3)2+ and Ru(phen)(3)2+ (bpy = 2,2'-bipyridine, and phen = 1,10-phenathroline) formed by silver ions of ground state in propylene carbonate solution are identified and characterized. Formation constants, lifetime, radiative and non-radiative decay constats of exciplexes are determined. Temperature-dependent luminescence lifetime measurements have revealed that exciplex formation causes a decrease in the energy level of the luminescent excited state. PMID- 11379017 TI - [Trends in functional mimicking of metallophosphatases]. AB - The promising practical applications of the so called artificial nucleases have inspired widespreading research studies on the functional mimicking of phosphoesterases. This paper is a short summary of the strategies for the selection or design of suitable model compounds, which are derived largely from the mechanistic details of phosphoesterases. PMID- 11379018 TI - [Preparation and spectroscopic studies of diorganotin(IV) complexes with adenosine and related compounds]. AB - Nine complexes of adenosine and related compounds (adenosine-5'-monophosphate, adenosine-5'-triphosphate, 1-methyl-adenosine, pyridoxal-5-phosphate and beta nicotinamide-adenine-dinucleotide-phosphoric acid) with di-n-butyltin(IV) oxide and/or di-n-butytin(IV) dichloride were prepared in the solid state. The compositions of the complexes were determined by standard analytical methods. It was found that the complexes contain organotin(IV) moiety and the ligand in a ratio of 1:1 or 2:1. The FTIR spectra demonstrated that di-n-butyltin(IV) oxide reacts with the D-ribose moiety of the ligands, while di-n-butyltin(IV) dichloride is co-ordinated to the deprotonated phosphate group. The basic part of the ligands does not participate directly in complex formation. Comparison of the experimental Mossbauer delta E values with those calculated on the basis of the PQS concept revealed that the organotin(IV) moiety has trigonal-bipyramidal, octahedral and in some cases tetrahedral geometry also. Some of the complexes contain the organotin(IV) cation in two different surroundings. PMID- 11379019 TI - [Estimation of impurity profiles of drugs and related materials. 20. Methodological problems in the identification and determination of organic impurities]. AB - After a brief survey of some fundamental questions related to the impurity profiling of drugs a scheme is presented for the rational estimation of the impurity profile of related organic impurities in drugs and drug products. In the course of the discussion of this scheme at first the methodological questions of the detection of the impurities are summarized followed by the description of their attempted identification with known potential impurities by means of chromatographic retention matching. A complex chromatographic (electrophoretic)- spectroscopic system is then described for the identification (structure elucidation) of those impurities which could not be identified by chromatographic retention matching. The first step of this is to draw conclusions from the UV spectra easily obtainable by the HPLC/diode array UV or TLC/reflection UV spectra. This is followed by the application of various on-line and off-line possibilities of mass spectrometry with special respect to the most up-to-date HPLC/MS/(MS) technique and concluded by the use of the ultimate and most informative techniques of NMR spectroscopy. Finally the necessity of the synthesis of the identified impurities (impurity standard) is briefly mentioned followed by the discussion of the necessity and possibilities of the selective determination of the identified impurities based on the impurity standards. PMID- 11379020 TI - [High performance liquid chromatographic enantiomer separation of beta-blocking agents with a new isothiocyanate type chiral derivatizing agent]. AB - The applicability of (1R,2R)-1,3-diacetoxy-1-(4-nitrophenyl)-2-propyl isothiocyanate [(R,R)-DANI] as a recently developed chiral derivatizing agent for the enantioseparation of a series of beta-blockers is described. The thiourea diastereomers formed were analysed by reversed-phase high-performance liquid chromatography, mixtures of water and methanol or acetonitrile being used for elution. Conditions of derivatizations (temperature, reagent excess and reaction time) were optimized. The effects of organic modifiers on the retention and separation were investigated; the diastereomers could readily be baseline separated with methanol-containing mobile phases. The reagent allowed the detection of all four stereoisomers of labetalol, which has two chiral centres. PMID- 11379021 TI - [Modeling of potential metal binding sites of glycoproteins]. AB - The metal ion coordination of carbohydrate type molecules is gradually influenced by the anchoring groups of the ligand molecules. Below we summarize our knowledge on the copper(II) complexes of the model compounds containing also carboxylate, ammine or both groups beside the alcoholic hydroxy groups obtained by combined potentiometric--UV/VIS, circular dichroism, electron spin resonance and nuclear magnetic resonance spectroscopic methods. PMID- 11379022 TI - [Investigation of protonation and complex formation processes of some new polyamino carboxylic acids]. AB - The medicinal chemistry often needs chemical compound with special properties. The preparation of compounds with special properties generally needs 10-20 steps of preparation and the final product is available in small amount. The characterisation of this kinds of compounds needs special methods and procedures for investigation. This kind of method has been developed and used for characterisation of some new polyaminopolycarboxylic acid derivatives. Protolytic equilibrium of 2,2',2",2'"-[terpyridine 6,6"diyl)bis(methylenenitrilo)]tetrakisacetic acid and 3,5-bis[6'-bis (N,N bis(carboxymethyl)aminomethyl]-2'-pyridyl)-1H,2, 4-triazole was investigated by pH-metric and spectrophotometric methods. The pK values of the aromatic nitrogen atoms were estimated. Complexation of Eu(III)ion with both ligands are studied by spectrophotometric method. Composition(ML2) and stability of complexes were determined. PMID- 11379023 TI - [Synthesis and in vitro receptor analysis of new potential oxytocin antagonists]. AB - Oxytocin (OT) itself and the sensitivity of the uterus to OT play a crucial role in the initiation of both normal and pathologically early delivery. It recognition of the complex mechanism of action suggests that specific OT antagonists are of therapeutic value in postponing early contractions. In this study neurohypophyseal hormone analogues containing Sar in position 7, Arg in position 8, and various conformationally restricted or bulky derivatives of phenilalanine and tryptophan amino acids in position 2 were prepared to design more potent and selective OT antagonists. We determined the ligand-receptor binding characteristics of these newly synthetized peptides in the presence of [3H]oxytocin and [3H]vasopressin on isolated guinea-pig uterus, rat liver and kidney inner medulla plasma membranes. In the case of each peptides we calculated the Ki values and the selectivity ratio. The binding to the OT receptor was dramatically decreased for the Trp-derivatives containing analogues, while the Phe-derivatives containing analogues displayed a relatively high receptor affinity and have a relatively high OT/VP1 receptor selectivity. PMID- 11379024 TI - [Speciation of bioactive Al(III) and VO(IV) complexes in biological systems]. AB - The speciation of the toxic Al(III) and the beneficial VO(IV) in various biofluids and tissues is discussed in order to describe the solution state of these metal ions in the organism. The possibility and the importance of ternary complex formation with relevant biomolecules is emphasised. The importance of the biospeciation of Al(III) ion in its transport and involvement in neurological disorders, and of insulin mimetic VO(IV) complexes is discussed. PMID- 11379025 TI - [7-aza analogs of quinazolino-carboline alkaloids. Synthesis, structure, characterization]. AB - The synthesis, structure and characterization of 8-substituted-7-azarutaecarpines (2) is described. These compounds were prepared by Fischer indolization of 3 amino-2-(1-phenylhydrazonoethyl)-4(3H)-quinazolin-one (5), followed by cyclocondensation with a series of aliphatic or aromatic aldehydes and formic acid or a Vilsmeyer-Haack reagent. The stereochemistry of compounds (2) was investigated by 1H nmr spectroscopy. It was found that the 8-substituents assume a quasi-axial position on the flattened boat conformation of ring C of (2), with the exception of ortho substituted phenyl groups, which occupy quasi-equatorial position. Semiempirical MO calculations support these conformational preferences. PMID- 11379026 TI - [A glimpse into a new discipline: medicinal inorganic chemistry]. AB - Authors discuss some aspects of the medicinal application of metal complexes. They deal with anti-effective agents, bismuth anticulcer and gold antiarthritic drugs, and with agents that influence the NO balance in the cardiovascular system. Further, they concern with insulin mimetic vanadium compounds, metal complexes of superoxide dismutase activity, photodynamic therapy, and the treatment of copper deficiency. PMID- 11379027 TI - [Sources of error in the European Pharmacopoeia assay of halide salts of organic bases by titration with alkali]. AB - A short overview has been given by the authors on the titrimetric assay methods of halide salts of organic bases in the pharmacopoeias of greatest importance. The alternative procedures introduced by the European Pharmacopoeia Commission some years ago to replace the non-aqueous titration with perchloric acid in the presence of mercuric acetate have also been presented and evaluated. The authors investigated the limits of applicability and the sources of systematic errors (bias) of the strongly preferred titration with sodium hydroxide in an alcoholic medium. To assess the bias due to the differences between the results calculated from the two inflexion points of the titration curves and the two real endpoints corresponding to the strong and weak acids, respectively, the mathematical analysis of the titration curve function was carried out. This bias, generally negligible when the pH change near the endpoint of the titration is more than 1 unit, is the function of the concentration, the apparent pK of the analyte and the ionic product of water (ethanol) in the alcohol-water mixtures. Using the validation data gained for the method with the titration of ephedrine hydrochloride the authors analysed the impact of carbon dioxide in the titration medium on the additive and proportional systematic errors of the method. The newly introduced standardisation procedure of the European Pharmacopoeia for the sodium hydroxide titrant to decrease the systematic errors caused by carbon dioxide has also been evaluated. PMID- 11379028 TI - [Model studies on the transport processes of anticancer platinum complexes]. AB - Potentiometric, calorimetric, NMR and stopped-flow kinetic studies were performed on the palladium(II) complexes of thioether and/or nitrogen donor ligands. The ternary systems always contained a tridentate ligand (dien, dipic, terpy and dianions of dipeptides, GlyGly, GlyAla and GlyMet) and a monodentate thioether (AcMet). The stability constants of thioether complexes were obtained by indirect potentiometric measurements using uridine as a competitive ligand. The thermodynamic parameters revealed that selectivity of palladium(II) for thioether binding can be significantly influenced by the other donor atoms around the metal ion. [Pd(terpy)]2+, [Pd(dipic)]2+ and [Pd(GlyMet)] had the lowest affinity for thioether binding and it was explained by steric and electronic effects. Ternary complexes of nitrogen donors have higher thermodynamic stability constants than that of the thioether complexes, but rate constants of the substitution reactions revealed that the formation of thioether complexes is the faster reaction. As a consequence, the thermodynamic equilibrium state of a multicomponent system is characterized by the coordination of N-donors, which are formed via the existence of thioether bonded intermediates. PMID- 11379029 TI - [Coordination compounds in the natural environment]. AB - The paper gives a brief overview from the complexation of metals by different ligands in the natural environment and it shows by several examples how transport, biological availability and environmental influences of the metals are attached by these processes. PMID- 11379030 TI - [The possibilities of GC/MS and HPLC in the analysis of sugars and acids in natural matrices]. AB - A GC/MS procedure is described for the simultaneous quantitation of the minor and major constituents of several natural matrices, including fruits, vegetables, mushrooms, drug/industrial plants and honeys, as their trimethylsilyl derivatives, from one solution, by one injection. Selected minor components have been determined on the basis of their characteristic fragment ions, in the presence of extremely high excess of saccharides of various degree of polymerization. Selective fragmentation of these minor compounds in the ion trap detector provided possibilities for distinguishing them. In the case of honeys the method permitted the simultaneous quantitation of o-phosphoric, malic, shikimic, citric/isocitric, quinic, margaric, oleic and stearic acids, 5 hydroxymethylfurfurol and proline with the extremely high sugar contents of honeys (fructose, glucose, galacturonic acid, inositol, sucrose, trehalose, turanose, maltose, gentiobiose, isomaltose, raffinose, erlose, melezitose, maltotriose, panose, isomaltotriose) and allowed the fast evaluation of sugar and acid constituents of fifteen honeys from various floral and geological origin. Results revealed that (i) the minor components varied in the concentration range of 0.0001 to 0.43%, and, (ii) together with the saccharides of honeys made up the total of identified and determined constituents from 87.8% to 98.5%. Quantitative evaluation of the minor constituents was performed on the basis of their selective fragment ion values with an average reproducibility of 6.7% (RSD). PMID- 11379031 TI - [The role of halogenation in the electro-oxidation of halogenated anilines in acetonitrile as the solvent]. AB - The 4-halogeno-, 2,4-dihalogeno- and 2,4,6-trihalogenoanilines were the model substances of the research described. Acetonitrile being one of the most popular aprotic solvent was used. The halogeno monomers were chloro and bromoanilines. The electrodimerised substrate is stabilised by rejecting halogenide ion and protons. The rejected halogenide ions are oxidisable more easier compared to the substrate therefore the halogene formed substitutes both the substrate, and the dimer as well. The protons formed during the oxidation bind by the strong base monomer primary amine. Because of these not only a simple electrodimerisation will take place, but protonised monomers, higher halogenated monomers, and the corresponding dimer molecules are formed in the solution. PMID- 11379032 TI - [Kinetics and mechanisms of the reactions of transition metal complexes]. AB - With the objective of elucidating the mechanisms of dioxygen activation by transition metal complexes, we have investigated functional models of some metalloenzymes, such as phenoxazinone synthase and catechol oxidase, based on bisdimethylglyoximatocobalt(II) and -iron(II) complexes. Kinetic studies and the identification of intermediates permitted to establish a free radical mechanism. The catalytic cycle involves H-atom abstraction from the catechol by a superoxo complex, generating a semi-quinone anion radical, which coordinates to the metal, affording new complexes. Useful free-radical intermediates in biological systems are not impossible since tyrosinase has been shown to contain coordinated tyrosyl radical as an integral part of the active enzyme. We have interpreted the structure-reactivity relationship observed in the reaction of the [Pd2Cl2(dppm)2] dimer with arenesulfonyl azides. PMID- 11379033 TI - [Preparation, X-ray structural and spectroscopic studies of some D-lactobionic acid complexes with Cs(I), Fe(III) and di-n-butyltin(IV)]. AB - D-Lactobionic acid (4-O-beta-D-galactopyranosyl-D-gluconic acid) complexes of Cs(I), Fe(III) and di-n-butyltin(IV)2+ ions were prepared in the solid state. The bonding sites of the ligands were verified by means of FTIR, Raman and 13C NMR spectroscopic measurements. The Cs(I)-D-lactobionate was obtained in single crystal form. The X-ray crystallographic results on Cs(I)-D-lactobionate demonstarted that each Cs(I) ion is bonded to four D-lactobionate ions, forming an intricate 3D network. The asymmetric unit consists of one Cs(I), one D lactobionate ion and one water molecule. For the di-n-butyltin(IV) complex, Mossbauer pqs calculations indicated octahedral and trigonalbipyramidal stereochemistry around the central tin atom in the oligomeric compound. In DMSO solution, the polymeric structure does not remain as shown by 13C NMR measurement. One solvent molecule is coordinated additionally to the tin center, and the carboxylate group has become monodentate. According to the EPR measurement, the Fe(III) complexes obtained at different pH have at least dimeric or oligomeric structure. PMID- 11379034 TI - [Positron lifetime studies of sodium- and zinc-hyaluronates]. AB - The aim of this work was to test the positron lifetime technique (PLT) as a tool of the structure study of sodium- and zinc-hyaluronates. The information based on the PLT measurements (outlined as follows) proved that this method can be useful in this field as well. The lifetime of ortho-positronium (o-Ps) significantly increased and its intensity decreased in the samples containing Zn2+, comparing to Na-hyaluronates. It showed that the electronic orbitals are more closed in the case of Zn2+ cation that means the overlapping between the wave functions of the positron and of the electrons decreased. (See equation No 1.) The study of the effect of water content suggested that the hydrogen-bridge-bonds "localised" the free electron pairs. The increasing pressure increased the lifetime and it is an evidence that the effect of the cations (Na+ and Zn2+) can be explained rather with the change of the electronic structure than with the altering of the free volumes of the samples. PMID- 11379035 TI - [Beta-cyclodextrin inclusion complexes: effect of the host substitution on molecular recognition]. AB - The stability of inclusion complexes (i.e. the measure of molecular recognition between a guest and beta-cyclodextrin) is highly influenced by the fit of the guest into the cavity of the host and by the secondary bonds among the functional groups getting in close connection. The strength of these interactions depend on the size, shape and functional groups of the guests as well as on the facts that both the size of the cavity and the reactivity of beta-cyclodextrins are altered when the hydroxy groups are substituted. As best models, the interactions among hydroxypropylated cyclodextrins of different average degree (and pattern) of substitution and phenolphthalein (as a model for "large" guests) and p nitrophenol-p-nitrophenolate couple (as for "small" ones) have been studied. The formation constants of phenolphthalein hydroxypropyl-beta-cyclodextrin complexes are continuously decreasing, while those of p-nitrophenol/phenolate ones show a maximum with increasing degree of substitution. The pattern of substitution has also a significant effect on the quality of the interactions: the substitution on O(6) position alters the type of the interactions most, and a series of different findings prove that this change is the basis in the chiral selectivity of different cyclodextrin derivatives, too. PMID- 11379036 TI - [Complexation properties of the ligand EOB-DTPA and the kinetic stability of Gd(EOB-DTPA)]. AB - Due to the presence of the ethoxy-benzyl group, the lipophilicity of the complex Gd(EOB-DTPA)2- is higher than that of Gd(DTPA)2- and thus Gd(EOB-DTPA)2- can be used as a liver specific contrast agent in Magnetic Resonance Imaging. The stability constants of the complexes of Fe2+, Fe3+, Zn2+ and Cu2+ formed with EOB DTPA practically agree with those of the respective DTPA complexes. The complexes of EOB-DTPA formed with larger metal ions, like Ca2+, Ba2+, Gd3+ and other lanthanides, exhibit larger stability constants than the complexes of DTPA by 0.5 1 logK unit. The stability constants of the complexes Ln(EOB-DTPA)2- increase with increasing atomic number till the middle of the series. For the elements Tb to Ho the logKLnL values are approximately equal and a weak decrease can be observed for the heaviest elements, probably because of steric hindrance between the functional groups. The exchange reactions between the complex Gd(EOB-DTPA)2- and Y3+ ions occur predominantly through the formation and rate determining dissociation of the mono- and diprotonated complexes. The rate constants characterizing these reaction pathways are lower by a factor of three than the respective rate constants, known for the dissociation of Gd(DTPA)2-. On the basis of its higher kinetic stability, Gd(EOB-DTPA)2- can be used for in vivo investigations in medical diagnosis. PMID- 11379037 TI - [Liberation of active agents from coherent emulsions]. AB - Drug release from coherent emulsions containing high water concentration (50-80 w/w%) was studied. Composition of coherent systems was as follows: self emulsifying wax and preserved water. Griseofulvin was applied as active agent in suspended form. The liberation experiments were carried out with Hanson vertical diffusion cell, acceptor phase was distilled water, membrane was celophane one. It was established that the time course of liberation of griseofulvin from coherent emulsions can be characterized with a multiplicative function and the exponent of this function is about 0.5. The quantity of released drug increased linearly with the water content and it decreased exponentially with the viscosity of coherent emulsions. PMID- 11379038 TI - Morphine-6 beta-glucuronide induces potent immunomodulation. AB - The effect of morphine administration on immune parameters is well documented. However, there exists a limited knowledge of the effect of morphine's metabolites on immune status. The present study examines the immunomodulatory effects of the morphine metabolite, morphine-6 beta-glucuronide (M6G), in the rat and provides further evaluation of the antinociceptive effects of M6G. Animals were administered phosphate-buffered saline (PBS) or M6G in doses of 1.0, 3.16, or 10.0 mg/kg (subcutaneous (s.c.)) or 0.1, 0.316, or 1.0 microgram (intracerebroventricular (i.c.v.)). Animals were tested for antinociception in the warm water tail-withdrawal procedure. In a separate set of animals, assessments of splenic natural killer cell activity, lymphocyte proliferative responses to mitogenic stimulation, and production of interferon-gamma were made 1 h following the s.c. or i.c.v. administration of M6G. The results show that M6G induced potent antinociception that was evident for at least 120 min following administration. M6G also produced decreases in natural killer cell activity, lymphocyte proliferation, and interferon-gamma production 1 h following both routes of administration. The difference in potency between immune alterations induced by subcutaneous vs. intracerebroventricular administration suggest central mediation of the immunomodulatory properties of M6G. Thus, M6G produces significant antinociception and immunomodulation in the rat. These findings demonstrate potent immunomodulatory properties of a metabolite of morphine, 1M6G. PMID- 11379039 TI - Effects of KE-758; an active metabolite of the new anti-rheumatic drug KE-298, D penicillamine, bucillamine and auranofin on the proliferation of murine lymphocytes, and the production of nitric oxide by murine macrophages. AB - 2-Mercaptomethyl-4-(4-methylphenyl)-4-oxobutanoic acid (KE-758), which is the active metabolite of 2-acetylthiomethyl-4-(4-methylphenyl)-4-oxobutanoic acid (KE 298), is a novel sulphydryl anti-rheumatic drug. In this study we analyzed the effect of KE-758 on the proliferation of murine lymphocytes, and on the production of nitric oxide (NO) by RAW264.7 murine macrophage cells. We compared its effect with other sulphydryl drugs such as D-penicillamine, bucillamine and auranofin. The proliferation of lymphocytes was measured by 3H-thymidine incorporation assay. Nitrite was measured using Griess Reagent. In the absence of copper ions, KE-758, D-penicillamine and bucillamine rarely affected the proliferation of concanavarin A (ConA) activated murine splenocytes. However, in the presence of copper, pharmacological concentrations of KE-758 but not D penicillamine and bucillamine suppressed the proliferation of murine splenocytes through a hydrogen peroxide-dependent mechanism. Auranofin markedly suppressed the proliferation regardless of the presence of copper ions by reducing the cellular viability. Furthermore, only KE-758 markedly suppressed the proliferation of phorbol myristate acetate (PMA) plus ionomycin activated murine whole blood lymphocytes (WBL) even in the absence of exogenous copper ions by a hydrogen peroxide-independent mechanism. Meanwhile, lipopolysaccharide (LPS) or LPS plus interferon-gamma (IFN-gamma) induced NO production by RAW264.7 cells were suppressed by KE-758 and auranofin but not by D-penicillamine and bucillamine. In conclusion, KE-758 is a novel immunosuppressive drug, which inhibits both lymphocyte and macrophage functions and its unique anti-rheumatic profile is distinct from that of D-penicillamine, bucillamine and auranofin. PMID- 11379040 TI - Immunomodulatory potential of hydrophobic analogs of Rigin and their role in providing protection against Plasmodium berghei infection in mice. AB - Here, we report the immunomodulating potential of N-palmitoyl-amino-ethyl-rigin amide (PR) and N-cholestanyl-amino-ethyl-rigin amide (CR), the two new structural analogs of rigin (an IgG-derived tetrapeptide). Their activity profiles are compared with native tuftsin (NT) and/or N-palmitoyl-amino-ethyl-tuftsin amide (PT) taken as positive control. To explore the possibility of their use as targeting molecules, they are incorporated into the liposome bilayer and, subsequently, interacted with macrophages in an in vitro study. The new analogs of rigin with the hydrophobicity introduced at the C-terminus are found to considerably improve both the cell-mediated and the humoral immune responses in mice. However, unlike tuftsin and its analog, which mainly activate polymorphonuclear leukocytes and macrophages, the rigin analogs appear to manifest their response more through lymphocytes. When administered prophylactically to a group of mice, at the dose of 100 micrograms/0.5 ml/mouse/day for 2 days (i.v.), followed by a challenge presented with 1 x 10(6) rbcs parasitised with Plasmodium berghei on day 0, substantial reduction in parasitaemia and rate of mortality is observed. This led to increase the median survival time (MST) of the treated group in comparison to the control group. The response is found to be more prominent in CR-treated mice possibly because of the presence of steroid moiety, which is likely to have more productive interaction with cell membranes. Incorporation of these peptides into the bilayer of liposomes does not alter the permeability behavior of vesicles and, in fact, enhances their uptake by the macrophages in an in vitro study. The effect, however, is dependent on both, the concentration of peptide liposomes and the time of incubation. Present study, thus, establishes the possible use of these analogs not only as adjuvant in chemotherapy, but also as a prophylactic supplement to boost the natural immune status. The activity response of rigin analogs is manifested through lymphocytes, they can also find use in the chemotherapy of diseases, like leishmaniasis, tuberculosis and leprosy, where macrophage activity is either tamed or impaired by pathogens. PMID- 11379041 TI - Dichotomous effect of a traditional Japanese medicine, bu-zhong-yi-qi-tang on allergic asthma in mice. AB - To determine the potentiality of prophylactic and/or therapeutic approaches using a traditional herbal medicine, Bu-zhong-yi-qi-tang (Japanese name: Hochu-ekki-to, HOT), for the control of allergic disease, we examined the effects of oral administration of HOT on a murine model of asthma allergic responses. When oral administration of HOT was begun at the induction phase immediately after OVA sensitization, eosinophilia and Th2-type cytokine production in the airway were reduced in OVA-sensitized mice following OVA inhalation. The serum levels of OVA specific immunoglobulin (Ig)E and IgG1 were significantly decreased, whereas the level of OVA-specific IgG2a was increased. Interleukin (IL)-4 production by spleen T cells in response to OVA was significantly suppressed, while Interferon (IFN)-gamma production was increased in mice treated with HOT in the induction phase. On the other hand, HOT given in the eliciting phase induced a predominant Th2 response with increased IgE production in OVA-sensitized mice following OVA inhalation. These results suggest that the oral administration of HOT dichotomously modulates allergic inflammation in a murine model for asthma, thus offering a different approach for the treatment of allergic disorders. PMID- 11379042 TI - Analysis of gene expression induced by irritant and sensitizing chemicals using oligonucleotide arrays. AB - Chemical-induced allergy continues to be an important occupational health problem. Despite decades of investigation, the molecular mechanisms underlying chemical-induced hypersensitivity and irritancy remain unclear because of the complex interplay between properties of different chemicals and the immune system. In this study, gene expression induced by toluene diisocyanate (TDI, a primarily IgE-inducing sensitizer), oxazolone (OXA, a cell-mediated hypersensitivity inducing sensitizer), or nonanoic acid (NA, a non-sensitizing irritant) was investigated using gene arrays. Female BALB/c mice were dermally exposed on the ears once daily for 4 consecutive days. On day 5, the lymph nodes draining the exposure sites were collected and used for RNA extraction and subsequent hybridization to Affymetrix Mu6500 oligonucleotide arrays. Of the 6519 genes on the arrays, there were 44, 13, and 51 genes in the TDI-, OXA-, and NA exposed samples, respectively, that displayed a minimum of twofold change in expression level relative to the vehicle control. There were 32, 19, and 19 genes that were differentially expressed (with a minimum of twofold change) between TDI and OXA, TDI and NA, OXA and NA, respectively. The differentially expressed genes include immune response-related genes, transcriptional factors, signal transducing molecules, and Expressed Sequence Tags. Based on the gene array results, candidate genes were further evaluated using RT-PCR. There was only about 47% concordance between the gene array and RT-PCR results. PMID- 11379043 TI - Cellular and humoral adjuvant activity of lectins isolated from Korean mistletoe (Viscum album colaratum). AB - The adjuvant effect of lectins (KML-C) isolated from Korean mistletoe (Viscum album coloratum) on induction of humoral and cellular immune responses against keyhole limpet hemocyanine (KLH) was examined. When mice were immunized subcutaneously (s.c.) with KLH (20 micrograms/mouse) admixed with or without 50 ng/mouse of KML-C (KLH + KML-C), mice immunized with KLH + KML-C showed significantly higher antibody titers against KLH than those immunized with KLH alone, showing the highest titer 5 weeks after immunization. Furthermore, boost immunization with KLH + KML-C at 2-week interval elicited much higher activity than single immunization to enhance antibody responses against KLH. The assay for determining isotypes of antibodies revealed that KML-C augmented KLH-specific antibody titers of IgG1, IgG2a and IgG2b. The culture supernatants obtained from the splenocytes of mice treated with KLH + KML-C also showed a higher level of both KLH-specific Th-1 (IL-2 and IFN-gamma) and Th-2 type cytokine (IL-4). In an in vitro analysis of T lymphocyte proliferation to KLH on week 4, the splenocytes of mice treated with KLH + KML-C showed a significantly higher proliferating activity than those treated with KLH alone. In addition, mice immunized twice with KLH + KML-C and followed by intrafootpad (i.f.) injection of KLH (50 micrograms/site) 14 weeks after the primary immunization induced a higher delayed type hypersensitivity (DTH) reaction than mice treated with KLH alone. These results suggest that KML-C is a potent immunoadjuvant to enhance cellular and humoral immune responses. PMID- 11379044 TI - Anti-allergy properties of fermented foods: an important immunoregulatory mechanism of lactic acid bacteria? AB - Clinical reports have suggested that dietary consumption of fermented foods, such as yogurt, can alleviate some of the symptoms of atopy and might also reduce the development of allergies, possibly via a mechanism of immune regulation. Controlled studies have indicated that consumption of fermented milk cultures containing lactic acid bacteria (LAB) can enhance production of Type I and Type II interferons at the systemic level. In animal models, LAB have been shown to promote interferon expression, and to reduce allergen-stimulated production of IL 4 and IL-5 in some cases. Recent results have shown that LAB are potent inducers of pro-interferon monokines (IL-12 and IL-18), and that cytokine secretion is stimulated by the interaction of Gram-positive cell wall components with surface receptors of mononuclear phagocytes, via NF-kappa B and STAT signalling pathways. However, it is clear that the extent and quality of LAB-induced immunoregulation is strain-dependent. This review discusses the clinical and laboratory evidence for anti-allergy properties of fermented foods, and proposes a model for the mechanism by which some well-defined strains of immunoregulatory LAB might down regulate a Th2 allergic phenotype. PMID- 11379045 TI - Parasympathetic modulation of amylase secretion by IFN gamma in murine submandibular glands. AB - IFN gamma is a pleiotropic cytokine that exerts immunologic and non-immunologic functions. We show here that at low doses (10 U/ml), it stimulates amylase secretion in murine submandibular glands (SMG) "via" muscarinic receptor activation, comparable to that produced by the muscarinic agonist carbachol. Both effects are blocked by atropine. NG-monomethyl-L-arginine (L-NMMA) and EGTA inhibited the cytokine effect on amylase secretion, involving the participation of a calcium-dependent isoform of nitric oxide synthase (NOS). We confirm NOS activation because IFN gamma stimulates nitrite production and enzyme activity in SMG. Carbachol (10(-7) M) did not modify basal nitric oxide production. In addition, both IFN gamma and carbachol increase prostaglandin E2 production in SMG, but while indomethacin potentiates IFN gamma effect on amylase secretion, it blunted amylase secretion exerted by carbachol. Thus, IFN gamma and carbachol stimulate IFN gamma secretion on SMG in a dose-dependent manner. Our results are pointing to neuroregulatory functions of IFN gamma in murine SMG, because it regulates its own levels in oral cavity, perhaps to exert a local immuno surveillance. PMID- 11379046 TI - Comparison of the effect of lidocaine-epinephrine and prilocaine-felypressine to alter macrophage functions. AB - In vitro treatment of macrophages with lidocaine-epinephrine or prilocaine felypressine resulted in inhibition of their adhesion, chemotaxis and phagocytosis. However, prilocaine-felypressine was a much more potent inhibitor of adhesion and phagocytosis than lidocaine-epinephrine. On the other hand, lidocaine-epinephrine induced transient potentiation of superoxide anion production by macrophages, while prilocaine-felypressine consistently inhibited this. Moreover, lidocaine-epinephrine and prilocaine-felypressine both inhibited the production of hydrogen peroxide. In contrast, epinephrine strongly potentiated superoxide anion production, while markedly inhibiting hydrogen peroxide production. This potentiation by epinephrine was not prevented by adrenergic antagonists. In addition, superoxide dismutase potentiated the production of hydrogen peroxide, which was in part prevented by epinephrine. These results suggest that lidocaine-epinephrine and prilocaine-felypressine inhibit adhesion, chemotaxis, phagocytosis, and the production of hydrogen peroxide by macrophages. In addition, lidocaine-epinephrine evidently differs from prilocaine-felypressine regarding the molecular mechanisms underlying the modulation of superoxide anion production by macrophages. PMID- 11379047 TI - Immunotoxicological investigation of subacute combined exposure by permethrin and the heavy metals arsenic(III) and mercury(II) in rats. AB - Effects of combined 28 days of oral exposure to the insecticide Permethrin (Pe), alone or in combination with arsenic-III (As) or Hg-II (Hg), were investigated on certain toxicological (body weight, organ weights), haematological (white blood cell (WBC) and red blood cell (RBC) counts, haematocrit (Ht), mean cell volume (MCV), cell content of the femoral bone marrow) and immune function (IgM-PFC, delayed-type hypersensitivity (DTH) reaction) parameters of male Wistar rats. Immunotoxic (H = high) and NOEL (L = low) doses of the three substances were determined in preliminary experiments under identical experimental conditions. In the present study, the immunotoxic dose of Pe (126 mg/kg) was combined with the NOEL dose of As (3.33 mg/kg) or Hg (0.40 mg/kg), and the NOEL dose of Pe (12.6 mg/kg) with the immunotoxic dose of As (13.3 mg/kg) or Hg (3.20 mg/kg). A separate group of animals, treated with the appropriate high dose component only, was used as internal control. Significant interactions were observed in the liver weight of the animals treated with Pe(H)-As(L) or As(H)-Pe(L), in the cell content of the femoral bone marrow in case of Pe(H)-As(L) and Pe(H)-Hg(L) combinations, as well as in the number of PFCs formed from 10(6) spleen cells in the Pe(H)-As(L) and in the maximum of DTH reaction in the Hg(H)-Pe(L) combination. The results show that combined exposures by the investigated substances modify the toxic (including immunotoxic) effects of the single compounds. These findings rise the probability that the interactions observed can also be present in human situations altering the health hazard of this three chemicals. PMID- 11379048 TI - Effects of the aqueous extract of epimedii herba on the antibody responses in mice. AB - Antibodies and cytokines in serum were detected in male ICR mice treated with the aqueous extract of Epimedii Herba (AEEH) at doses of 40, 120 and 360 mg/kg orally for 2 weeks. Effects of AEEH on antibody forming responses were assessed by enzyme linked immunosorbent assay (ELISA) of immunoglobulin (Ig) levels in serum collected 7 days after priming with ovalbumin (OVA) in complete Freund's adjuvant (CFA) or immediately without priming at week 2. The relative spleen weight was significantly increased by AEEH, compared with controls, especially at a dose of 120 mg/kg of it after priming with OVA and 40 mg/kg without priming, respectively. However, body weight gain was slightly decreased in AEEH-fed mice. The enhancement of total serum IgG and IgG1 levels in unprimed mice was statistically significant in mice fed 40 mg/kg AEEH. Total serum IgG2a levels and Il-4 secretion were also statistically augmented by all groups of AEEH treatment. A tendency to marked increase of total serum IgM level and IFN-gamma secretion was also observed in mice fed 40 and 120 mg/kg AEEH but not those fed 360 mg/kg AEEH. When mice were immunized with OVA, furthermore, a marked stimulation of antibody formation and cytokines secretion was observed in all groups of AEEH-fed mice compared with controls. These findings indicate that AEEH at therapeutic concentrations enhances the production of antibodies and cytokines in mice, and the enhancing effects are more marked when the mice were immunized with OVA. Thus, these results suggest that AEEH is effective on Th cell functions, and protective effects on host against immune diseases. PMID- 11379049 TI - Effect of the antianginal drug bepridil on intracellular Ca2+ release and extracellular Ca2+ influx in human neutrophils. AB - To understand more fully the effects of bepridil, an antiarrhythmic and antianginal drug, on myocardial ischemia-reperfusion injury and systemic immune responses, its effect on intracellular Ca2+ levels ([Ca2+]i) in human neutrophils was investigated by using fura-2 as a fluorescent probe. Bepridil (10-200 microM) increased [Ca2+]i in a concentration-dependent fashion. This signal was partly inhibited by removal of extracellular Ca2+. In a Ca(2+)-free medium, pretreatment with bepridil (100 microM) abolished the Ca2+ release induced by thapsigargin (1 microM), an endoplasmic reticulum Ca2+ pump inhibitor, and by carbonylcyanide m chlorophenylhydrazone (2 microM), a mitochondrial uncoupler. Pretreatment with carbonylcyanide m-chlorophenylhydrazone and thapsigargin, respectively, partly inhibited bepridil-induced Ca2+ release. Addition of Ca2+ (3 mM) increased [Ca2+]i after pretreatment with bepridil (100 microM) in a Ca(2+)-free medium. Bepridil (100 microM)-induced Ca2+ release was not altered when phospholipase C was inhibited by U73122 (2 microM). Both Ca2+ release and Ca2+ entry induced by bepridil (100 microM) were augmented by activating protein kinase C with phorbol 12-myristate 13-acetate (10 nM), and were suppressed by inhibiting protein kinase C with GF 109203X (2 microM). Treatment with bepridil (10-20 microM) for 30 min increased the production of reactive oxygen intermediates (ROI) by more than 50%. Collectively, it was found that bepridil increased [Ca2+]i concentration dependently in human neutrophils by releasing Ca2+ from the endoplasmic reticulum, mitochondria and, possibly, other compartments in a phospholipase C independent manner. Bepridil also activated Ca2+ influx. The activity of protein kinase C may regulate bepridil-induced Ca2+ release and Ca2+ entry. PMID- 11379050 TI - Functional studies of bradykinin receptors in Chinese hamster ovary cells stably expressing the human B2 bradykinin receptor. AB - Bradykinin B1 and B2 receptors, members of the G-protein coupled receptor superfamily, are involved in inflammation and pain. Chinese hamster ovary (CHO) cells stably expressing the human B2 bradykinin receptor (CHO-B2) were used to characterize the signal transduction pathways associated with this receptor and its regulation. The selective B2 antagonist [3H]NPC17731 but not the selective B1 antagonist [3,4-prolyl-3,4-(3)H(N)]-[des-Arg10,Leu9]kallidin ([3H]DALKD) bound to CHO-B2 cell membranes with a Kd of 0.77 nM and a Bmax of 1087 fmol/mg protein. [3H]NPC17731 binding was inhibited by bradykinin ligands in the order: NPC17731 > bradykinin > kallidin >> DALKD > [des-Arg10] kallidin (DAKD), consistent with the pharmacological profile of B2 bradykinin receptors. The B2 agonist bradykinin and the B1/B2 agonist kallidin, but not the B1 agonist DAKD, increased [35S]GTP gamma S binding to the CHO-B2 cell membranes. The B2 bradykinin receptors were co immunoprecipitated with G alpha q/11. In response to bradykinin stimulation, coupling of the B2 receptors to G alpha q/11 was increased by 10-fold. Bradykinin and kallidin, but not DAKD, induced intracellular calcium release in CHO-B2 cells, which was blocked by NPC17731 but not by DALKD. These results demonstrate that B2 bradykinin receptors directly coupled to G alpha q/11 to regulate intracellular calcium release. CHO-B2 cell is a useful system that can be applied to study the effect of potential agents that may influence the B2 receptor function. PMID- 11379051 TI - Sodium nitroprusside induces apoptosis of H9C2 cardiac muscle cells in a c-Jun N terminal kinase-dependent manner. AB - Sodium nitroprusside (SNP) induces apoptosis in H9C2 cardiac muscle cells. Treatment with an exogenous NO donor SNP (2 mM) to H9C2 cells resulted in apoptotic morphological changes; a bright blue-fluorescent condensed nuclei and chromatin fragmentation by fluorescence microscope of Hoechst 33258-staining. The activity of caspase-3 like protease was increased during SNP-induced cell death. However, the activity of caspase-1 like protease was not affected by SNP. Pretreatment with Z-VAD-FMK (a pan-caspase inhibitor) or Ac-DEVD-CHO (a specific caspase-3 inhibitor) abrogated the SNP-induced cell death. SNP markedly activated three MAP kinases (JNK/SAPK, ERK and p38 MAP kinase) in the cardiac muscle cells. In this study, selective inhibition of the ERK or p38 MAPK pathway (by PD98059 or SB203580, respectively) had no effect on the extent of SNP-induced apoptosis in cardiac muscle cells. In contrast, inhibition of the JNK pathway by transfection of a dominant negative mutant of JNK markedly reduced the extent of SNP-induced cell death. Taken together, we suggest that JNK/SAPK will be related to SNP induced apoptosis of H9C2 cardiac muscle cells. PMID- 11379052 TI - Diagnosis and management of specific breast abnormalities. PMID- 11379053 TI - Developmental neurotoxicity of methanol exposure by inhalation in rats. AB - The possibility of widespread methanol exposure via inhalation stemming from its adoption as an automotive fuel or fuel component arouses concerns about the potential vulnerability of the fetal brain. This project was designed to help address such concerns by studying the behavior of neonate and adult rats following perinatal exposure to methanol vapor. Four cohorts of pregnant Long Evans hooded rats, each cohort consisting of an exposure and a control group, were exposed to 0 parts per million (ppm) (control) or 4,500 ppm methanol vapor for six hours daily beginning on gestation day (GD) 6 with dams and pups then being exposed postnatal day (PND) 21. Exposures took place in 2-m3 Rochester-type inhalation chambers while the animals remained in their plastic breeder cages. Prenatal and postnatal blood methanol concentrations were determined by gas chromatography. Blood methanol concentrations of the dams, measured immediately following a six-hour exposure, were approximately 500 to 800 micrograms/mL throughout gestation and lactation. Average blood methanol concentrations of the pups were about twice those of the dams. Because such results appeared consistently across the other cohorts, we decided to obtain additional data with Cohort 4. Once it had undergone the standard exposure protocol, we selected sets of extra pups from those that had not been assigned previously to the adult phase of behavioral testing. Each set was exposed once, at ages that extended out to PND 52, for one additional six-hour session of exposure to 4,500 ppm methanol. The blood methanol concentrations of these pups declined until about PND 48, at which time they approximated those of the dams. These findings might be accounted for by a process of metabolic maturation in the pups that remains to be identified. PMID- 11379054 TI - Activation of eicosanoid metabolism in human airway epithelial cells by ozonolysis products of membrane fatty acids. AB - Inhaled ozone can react with a variety of cellular macromolecules within the lung. Recent analyses of the chemistry of ozone reactions with unsaturated fatty acids, which are present in all membranes and in mucus in the airways, indicate that ozonolysis yields one aldehyde and one hydroxyhydroperoxide molecule for each molecule of ozone. The hydroxyhydroperoxide molecule is unstable in aqueous environments, and subsequently yields a second aldehyde and hydrogen peroxide. The structure of common unsaturated fatty acids is such that attack by ozone at the carbon-carbon double bonds will yield 3-, 6-, and 9-carbon saturated and unsaturated aldehydes and hydroxyhydroperoxide. This study examines the effects of ozonolysis products on eicosanoid metabolism in human airway epithelial cells. Eicosanoid biosynthesis is important in a wide array of pathophysiological responses in the airway, and the release of eicosanoids by the epithelial barrier is likely to be significant in diseases induced by environmental factors. Previously, we demonstrated that ozone can increase eicosanoid synthesis from airway epithelial cells exposed in vitro. Human exposures to concentrations of ozone below the current National Ambient Air Quality Standard (0.12 ppm, not to be exceeded for more than one hour once per year) also resulted in increased eicosanoids in bronchoalveolar lavage fluid. To determine whether ozonolysis products could activate eicosanoid release, we exposed human airway epithelial cells to 3-, 6-, and 9-carbon aldehydes, hydroxyhydroperoxides, and hydrogen peroxide. We measured (1) eicosanoid metabolism using high-performance liquid chromatography and radioimmunoassays, and (2) the effects of the aldehydes, hydroxyhydroperoxides, and hydrogen peroxide on cell lysis. Eicosanoid release increased after exposure to aldehyde; release induced by 9-carbon (nonanal) aldehyde was greater than that induced by the 6-carbon (hexanal) or 3-carbon (propanal) aldehydes. Hydroxyhydroperoxides induced greater eicosanoid release than the corresponding aldehydes of equivalent chain length. Again, the longer the aliphatic chain length of the hydroxyhydroperoxide the greater the effect. These effects were noted at concentrations of hydroxyhydroperoxide below those that produce cell lysis, and the time course of the two responses was dissimilar. Because hydroxyhydroperoxides can degrade into an aldehyde and hydrogen peroxide, it is conceivable that the effects observed were attributable to the formation of either hydrogen peroxide or hydrogen peroxide and aldehyde. This mechanism is unlikely, however, because the effects of hydroxyhydroperoxides on eicosanoid release were dependent on chain length, whereas each hydroxyhydroperoxide can produce only one hydrogen peroxide molecule. Although hydrogen peroxide alone also stimulated eicosanoid metabolism, this effect was not augmented when aldehyde and hydrogen peroxide were added together. In addition, the dose of hydroxyhydroperoxide needed to produce an effect (10 to 100 microM) was lower than that of hydrogen peroxide (300 microM). We could not fully evaluate the effects of the unsaturated aldehydes and hydroxyhydroperoxides. Although the 6 carbon and 9-carbon cis-3-aldehydes could be synthesized from the cis-3-alcohols, the resulting aldehydes were not chemically stable. The cis-3-aldehydes were useful for producing the corresponding 1-hydroxy-alkenyl-hydroperoxides of high purity. These results support the method selected for chemical synthesis, but further studies are required to establish proper storage and handling methods before these compounds can be tested in assays of eicosanoid metabolism. PMID- 11379055 TI - Hot particles--a challenge within radioecology. PMID- 11379056 TI - Special issue on environmental radiometrics. PMID- 11379057 TI - Mapping 137Cs deposition: data validation methods and data interpretation. AB - The "Atlas of caesium deposition on Europe after the Chernobyl accident" was prepared within the framework of the Joint Study Project 6 (JSP6) of the collaborative programme on the consequences of the Chernobyl accident between the European Commission and the Ministries responsible for Chernobyl affairs in Belarus, Russia and Ukraine. The radiological data provided by scientific institutes and competent authorities of more than 30 European countries were integrated into an information platform. Data validation and intercomparison were therefore essential before the interpretation of the deposition with the help of isoline maps prepared with a geographic information system (GIS) could be done. The data validation was a two-step procedure: after performing a primary logical consistency check, other techniques based on spatial statistics were used to outline uncertain data. The purpose of this paper is to present these validation methods and to discuss the advantages and constraints of these techniques. Rather than trying to improve the techniques of spatial data analysis, suggestions are made on how an adaptation of the sampling information could improve the final result. PMID- 11379058 TI - Airborne surveys of Swiss nuclear facility sites. AB - Annually since 1989, biannually since 1994, the environs (approximately 50 km2) of the Swiss nuclear facilities are surveyed (by helicopters) flying the same survey lines. The equipment and the data processing software used for these surveys were built and developed at the Institute of Geophysics, ETH Zurich. For mapping of man-made radiation at or around nuclear facility sites a pixel representation and the MMGC (man-made gross count) ratio is used. So far no artificial radioactivity that could not be explained by the Chernobyl event (1986) or by earlier nuclear weapon tests was detected outside the fenced sites of the nuclear facilities. PMID- 11379059 TI - Aerial measurements on uranium ore mining, milling and processing areas in Germany. AB - The paper describes aerial measurements at several sites containing residues from uranium mines, waste rock dumps and tailings ponds of the Wismut company in Saxony and Thuringia. For the measurements, a computerized gamma-ray spectrometer equipped with a HPGe-detector and a NaI(Tl)-detector array with a volume of 121 was used. Radioactive anomalies from natural radionuclides have been detected and mapped. The activity concentrations of 226Ra, 228Th and 40K in areas with elevated radioactivity from natural radionuclides were determined from aerial measurements. Ground-based measurements--as far as available--confirmed these results. The mean 226Ra activity concentrations in the different waste rock dumps were found to be in the range 370 to 1600 Bq kg-1. The highest mean 226Ra activity concentrations were measured for the tailings ponds with values up to 1300 Bq kg-1. For 40K and 208Tl, activity concentrations of 860 and 40 Bq kg-1, respectively, were obtained as averages of all measurements. These values are commonly found in the environment. In general, there is reasonable agreement between the results measured with different detectors and measuring methods. PMID- 11379060 TI - Development and application of marine gamma-ray measurements: a review. AB - The development of instruments to measure gamma radiation in the marine environment, particularly on the sea floor, and the range of uses to which they have been put is reviewed. Since the first steps in the late 1950s, systems have been developed in at least 10 countries with the main thrust occurring in the 1970s. Development has continued up to the present, primarily in Europe and the USA. Marine gamma-ray spectrometers have been used for a range of applications including the mapping of rocks and unconsolidated sediments, mineral exploration (mainly for heavy minerals and phosphorites), sediment transport studies and investigations in relation to discharged and dumped nuclear wastes and at nuclear weapon test sites. PMID- 11379062 TI - Spectral deconvolution and operational use of stripping ratios in airborne radiometrics. AB - Spectral deconvolution using stripping ratios for a set of pre-defined energy windows is the simplest means of reducing the most important part of gamma-ray spectral information. In this way, the effective interferences between the measured peaks are removed, leading, through a calibration, to clear estimates of radionuclide inventory. While laboratory measurements of stripping ratios are relatively easy to acquire, with detectors placed above small-scale calibration pads of known radionuclide concentrations, the extrapolation to measurements at altitudes where airborne survey detectors are used bring difficulties such as air path attenuation and greater uncertainties in knowing ground level inventories. Stripping ratios are altitude dependent, and laboratory measurements using various absorbers to simulate the air-path have been used with some success. Full scale measurements from an aircraft require a suitable location where radionuclide concentrations vary little over the field of view of the detector (which may be hundreds of metres). Monte Carlo simulations offer the potential of full-scale reproduction of gamma-ray transport and detection mechanisms. Investigations have been made to evaluate stripping ratios using experimental and Monte Carlo methods. PMID- 11379061 TI - Seabed gamma-ray spectrometry: applications at IAEA-MEL. AB - The technique of underwater gamma-ray spectrometry has been developed to complement or replace the traditional sampling-sample analysis approach for applications with space-time constraints, e.g. large areas of investigation, emergency response or long-term monitoring. IAEA-MEL has used both high efficiency NaI(Tl) and high-resolution HPGe spectrometry to investigate contamination with anthropogenic radionuclides in a variety of marine environments. Surveys at the South Pacific nuclear test sites of Mururoa and Fangataufa have been used to guide sampling in areas of high contamination around ground zero points. In the Irish Sea offshore from the Sellafield nuclear reprocessing plant, a gamma-ray survey of seabed sediment was carried out to obtain estimates of the distribution and subsequently, for the inventory of 137Cs in the investigated area. PMID- 11379063 TI - Full-spectrum analysis of natural gamma-ray spectra. AB - In this paper, a new system to measure natural gamma-radiation in situ will be presented. This system combines a high-efficiency BGO scintillation detector with full-spectrum data analysis (FSA). This technique uses the (nearly) full spectral shape and the so-called 'standard spectra' to calculate the activity concentrations of 40K, 232Th and 238U present in a geological matrix (sediment, rock, etc.). We describe the FSA and the determination of the standard spectra. Standard spectra are constructed for various geometries and a comparison in intensity and shape will be made. The performance of such a system has been compared to a more traditional system, consisting of a NaI detector in combination with the 'windows' analysis. For count rates typically encountered in field experiments, the same accuracy is obtained 10-20 times faster using the new system. This allows for shorter integration times and hence shorter measurements or a better spatial resolution. The applicability of such a system will be illustrated via an example of an airborne experiment in which the new system produced results comparable to those of much larger traditional systems. This paper will conclude with a discussion of the current status of the system and an outlook for future research. PMID- 11379064 TI - Extraction of full absorption peaks in airborne gamma-spectrometry by filtering techniques coupled with a study of the derivatives. Comparison with the window method. AB - In this paper, an adaptation of a spectral profile analysis method, currently used in high-resolution spectrometry, to airborne gamma measurements is presented. A new algorithm has been developed for extraction of full absorption peaks by studying the variations in the spectral profile of data recorded with large-volume NaI detectors (16 l) with a short sampling time (2 s). The use of digital filters, taking into consideration the characteristics of the absorption peaks, significantly reduced the counting fluctuations, making detection possible based on study of the first and second derivatives. The absorption peaks are then obtained by modelling, followed by subtraction of the Compton continuum in the detection window. Compared to the conventional stripping ratio method, spectral profile analysis offers similar performance for the natural radioelements. The 137Cs 1SD detection limit is approximately 1200 Bq/m2 in a natural background of 200 Bq/kg 40K, 33 Bq/kg 238U and 33 Bq/kg 232Th. At low energy the very high continuum leads to detection limits similar to those obtained by the windows method, but the results obtained are more reliable. In the presence of peak overlaps, however, analysis of the spectral profile alone is not sufficient to separate the peaks, and further processing is necessary. Within the framework of environmental monitoring studies, spectral profile analysis is of great interest because it does not require any assumptions about the nature of the nuclides. The calculation of the concentrations from the results obtained is simple and reliable, since only the full absorption contributions are taken into consideration. A quantitative estimate of radioactive anomalies can thus be obtained rapidly. PMID- 11379065 TI - A code to simulate nuclear reactor inventories and associated gamma-ray spectra. AB - A computer code has been developed to simulate the gamma-ray spectra that would be measured by airborne gamma spectrometry (AGS) systems from sources containing short-lived fission products. The code uses simple numerical methods to simulate the production and decay of fission products and generates spectra for sodium iodide (NaI) detectors using Monte Carlo codes. A new Monte Carlo code using a virtual array of detectors to reduce simulation times for airborne geometries is described. Spectra generated for a short irradiation and laboratory geometry have been compared with an experimental data set. The agreement is good. Spectra have also been generated for airborne geometries and longer irradiation periods. The application of this code to generate AGS spectra for accident scenarios and their uses in the development and evaluation of spectral analysis methods for such situations are discussed. PMID- 11379066 TI - A European bibliography on airborne gamma-ray spectrometry. PMID- 11379067 TI - Determination of 228Ra, 226Ra and 224Ra in natural water via adsorption on MnO2 coated discs. AB - A fast procedure based on sorption of Ra on MnO2 coated polyamide discs is presented for determination of radium isotopes (i.e. 228Ra, 226Ra, 224Ra) in aqueous samples. The sample discs can be used directly for low-level alpha spectrometry without the need for further separation and preparation methods to produce planar sample sources. While the activity of alpha-emitting 224Ra and 226Ra can be determined during a first measurement, beta-emitting 228Ra is obtained via ingrowth of the progeny 228Th on the same sample disc after a standing time of about six months. Calculations are presented for optimizing the analytical accuracy as well as for predicting the sorption yield or chemical recovery of radium on the sample disc as a function of exposure time because the sorption uptake proceeds with first-order kinetics. The analyses can be carried out on small samples of 0.5-11 and, for long counting times of one week and use of high-purity silicon surface barrier detectors, a detection limit of 0.15 mBq l 1 is obtained for 226Ra. Since the half-life of 224Ra is only 3.7 d and since 228Th (as a measure for 228Ra) is built up only partially on the sample disc, a slightly higher detection limit of 0.24 mBq l-1 results for the latter isotopes. The procedure is therefore sufficiently sensitive to allow the investigation of Ra isotope relationships in aquifers at typical environmental levels. PMID- 11379068 TI - 228Ra/226Ra/224Ra and 87Sr/86Sr isotope relationships for determining interactions between ground and river water in the upper Rhine valley. AB - Ground and river waters of the upper Rhine valley (Alsace, France) were investigated for chemical composition of the major elements, Sr isotopes and radionuclides from the U and Th series. In particular, the isotope ratios and concentrations of Ra and Sr were used as geochemical tracers to distinguish between different types of water and their interactions. The bulk chemical analyses suggest that the surface waters in the Rhine valley can be described as mixtures between Ca-Na-HCO3-rich ground water and less mineralized slightly acidic river waters which have migrated through crystalline (mainly granitic) basement rocks of the Vosges mountains. Mixing of these waters yields positive correlation between bulk Sr, U, Ca and HCO3, indicating that carbonate-rich sediments are the main source of U and (non-radiogenic) Sr in the Rhine valley aquifers. The combination of the Ra and Sr isotope systems (228Ra/226Ra, 87Sr/86Sr) shows, however, that probably three sources contribute to the surface river waters in the upper Rhine valley, i.e. (i) a highly radiogenic crystalline component, (ii) a ground water source and, (iii) a third component from infiltrating Rhine water along the flow path of the parallel running river Ill in the northerly direction. The Sr and Ra isotope systems were also used to calculate small-scale mixing fractions of tributaries along the flow path of the Ill. Mixing ratios of non-pure end-member waters were determined using three isotope diagrams (i.e. 224Ra/226Ra vs. 228Ra/226Ra) and the results obtained with the Ra isotope system were found to be consistent with the data using Sr isotope relationships (i.e. 87Sr/86Sr vs. 1/Sr). PMID- 11379069 TI - Ground water input to coastal salt ponds of southern Rhode Island estimated using 226Ra as a tracer. AB - The naturally occurring radionuclide 226Ra (t1/2 = 1600 years) was used as a tracer to determine ground water input to Point Judith, Potter, Green Hill and Ninigret ponds in southern Rhode Island. Measurements of 226Ra activity were made in samples collected from salt ponds, pore waters, sediments, and local ground water wells during June-August, 1997. These results were combined with a simple box model to derive ground water input fluxes of 0.1-0.3 cm3 cm-2 d-1 (2-5 x 10(7) L d-1), which are comparable to previous estimates of ground water input to these ponds. PMID- 11379070 TI - 226Ra bioavailability to plants at the Urgeirica uranium mill tailings site. AB - Large amounts of solid wastes (tailings) resulting from the exploitation and treatment of uranium ore at the Urgeirica mine (north of Portugal) have been accumulated in dams (tailing ponds). To reduce the dispersion of natural radionuclides into the environment, some dams were revegetated with eucalyptus (Eucalyptus globolus) and pines (Pinus pinea). Besides these plants, some shrubs (Cytisus spp.) are growing in some of the dams. The objective of this study is to determine the 226Ra bioavailability from uranium mill tailings by quantifying the total and available fraction of radium in the tailings and to estimate its transfer to plants growing on the tailing piles. Plant and tailing samples were randomly collected and the activity concentration of 226Ra in plants (aerial part and roots) and tailings was measured by gamma-spectrometry. The exchangeable fraction of radium in tailings was quantified using one single step extraction with 1 mol dm-3 ammonium acetate (pH = 7) or 1 mol dm-3 calcium chloride solutions. The results obtained for 226Ra uptake by plants show that 226Ra concentration ratios for eucalyptus and pines decrease at low 226Ra concentrations in the tailings and appear relatively constant at higher radium concentrations. For shrubs, the concentration ratios increase at higher 226Ra solid waste concentrations approaching a saturation value. Percentage values of 16.0 +/- 8.3 and 12.9 +/- 8.9, for the fraction of radium extracted from the tailings, using 1 mol dm-3 ammonium acetate or calcium chloride solutions, respectively, were obtained. The 226Ra concentration ratios determined on the basis of exchangeable radium are one order of magnitude higher than those based on total radium. It can be concluded that, at a 95% confidence level, more consistent 226Ra concentration ratios were obtained when calculated on the basis of available radium than when total radium was considered, for all the dams. PMID- 11379071 TI - Exposure of workers in mineral processing industries in Brazil. AB - The mining, milling and processing of uranium and thorium bearing minerals may result in radiation doses to workers. A preliminary survey pilot program, that included six mines in Brazil (two coal mines, one niobium mine, one nickel mine, one gold mine and one phosphate mine), was launched in order to determine the need to control the radioactive exposure of the mine-workers. Our survey consisted of the collection and analysis of urine samples, complemented by feces and air samples. The concentrations of uranium, thorium and polonium were measured in these samples and compared to background data from family members of the workers living in the same dwelling and from residents from the general population of Rio de Janeiro. The results from the coal mines indicated that the inhalation of radon progeny may be a source of occupational exposure. The workers from the nickel, gold and phosphate mines that were visited do not require a program to control internal radiological doses. The niobium mine results showed that in some areas of the industry exposure to thorium and uranium might occur. PMID- 11379072 TI - Nuclide migration and the environmental radiochemistry of Florida phosphogypsum. AB - Phosphogypsum, a waste by-product derived from the wet process production of phosphoric acid, represents one of the most serious problems facing the phosphate industry in Florida today. This by-product gypsum precipitates during the reaction of sulfuric acid with phosphate rock and is stored at a rate of about 40 million tons per year on several stacks in central and northern Florida. The main problem associated with this material concerns the relatively high levels of natural uranium-series radionuclides and other impurities which could have an impact on the environment and prevent its commercial use. We have studied the potential release of radionuclides from phosphogypsum by: (i) analysis of stack fluids, groundwaters, and soils associated with gypsum stacks; and (ii) geochemical modeling. Stack fluids were observed to be very high in dissolved uranium and 210Pb with only moderate concentrations of 226Ra. Underlying soils tend to be enriched in U and 210Pb indicating precipitation when acidic stack fluids enter a buffered environment. Modeling results showed significant increases in radionuclide complexes with sulfate and phosphate, resulting in relatively mobile uncharged or negatively charged solution species within the stacks with likely precipitation of multicomponent solids with increasing pH below the stack. Our evidence thus suggests that, while phosphogypsum stacks do contain significant quantities of dissolved radionuclides, removal mechanisms appear to prevent large-scale migration of radionuclides to the underlying aquifer. PMID- 11379073 TI - Assessment of acid rock drainage pollutants release in the uranium mining site of Pocos de Caldas--Brazil. AB - We compared three different techniques to assess acid drainage occurrence connected to pyritic waste rock piles at a uranium mining and milling site in Pocos de Caldas--Brazil: (1) mass balance calculations, (2) column leaching experiments and (3) geochemical modelling. The study site was chosen because all the drainage coming from the pile is collected in one holding pond and a huge database (monitoring program) was available. The three independent methods predicted similar values for the intrinsic oxidation rate (IOR) (about 10(-9) kg m-3 s-1). We estimate the total time for consumption of all oxidizable material in the dump to be greater than 500 years. Geochemical model results showed a good agreement between predicted sulphate concentrations in relation to those found in the waste pile drainage, although the Al values were overestimated and pH values were underestimated. PMID- 11379074 TI - Radium contamination of the banks of the river Laak as a consequence of the phosphate industry in Belgium. AB - For over half a century, phosphate ores of marine origin, containing 226Ra, have been processed in Belgium to produce calcium phosphate for use in cattle food. As a result, the waste water containing 226Ra were discharged into two little rivers, one of which is the Laak. The purpose of this study was to chart the radium contamination of the river banks and some areas that are regularly flooded by the river. It was seen that enhanced concentrations of 226Ra do occur along the river banks, but that the contaminated area is mostly confined to a 10 m strip on both sides of the river, even in the flooding zones. At present, no dwellings are present on top of the contamination and no crops for direct human consumption are grown there, so there is no immediate threat to the population. PMID- 11379075 TI - Radionuclide fluxes at a plant manufacturing dicalcium phosphate for domestic animals. AB - We have studied a phosphate rock plant which produces dicalcium phosphate (DCP), used as a source of calcium and phosphorus for domestic animals. A by-product in the manufacturing process is calcium chloride which is used in the oil industry, the food industry and as road-salt. The objectives of our study were to describe the fluxes of radionuclides from the 238U decay series and to estimate the radiation doses to workers at the plant. The radionuclides in the phosphate rock were found to be in secular radioactive equilibrium with 238U, with an average activity concentration of 837 Bq kg-1. Separation and concentration processes were observed at different stages in the plant. Most of the 226Ra was found in the calcium chloride, while the major part of the 238U, about 950 Bq kg-1, was found in the dicalcium phosphate. The annual occupational effective dose to the workers was found to be below the 1 mSv limit recommended by ICRP (1991a) for the public. This study has shown a good example of an important non-nuclear industry with a high input of natural radionuclides with several conceivable pathways to man. PMID- 11379076 TI - Radiological impact from atmospheric releases of 238U and 226Ra from phosphate rock processing plants. AB - Phosphate rocks are used extensively, mainly as a source of phosphorus for fertilizers and secondarily for phosphoric acid and other speciality chemicals. Phosphates are typically enriched in uranium and are thus one of the sources of technologically enhanced natural radiation (TENR) which increases exposure to man from natural radionuclides. Emissions from phosphate rock processing plants in gaseous and particulate form contain radionuclides, such as 238U and 226Ra, which are discharged into the environment causing radiation exposures to the population. About 10 MBq each of 238U and 226Ra are discharged into the environment each year from SICNG, a phosphate rock processing plant in Thessaloniki area, Northern Greece. The collective dose commitment to lung tissue resulting from atmospheric releases was estimated to be approximately 2 x 10(-9) person Gy t-1 for 238U and approximately 0.1 x 10(-9) person Gy t-1 for 226Ra, i.e. about 2 times higher than that estimated in the UNSCEAR reports issued in 1982, 1988, and 1993. PMID- 11379077 TI - Contamination of settling ponds and rivers as a result of discharge of radium bearing waters from Polish coal mines. AB - Saline waters from underground coal mines in Poland often contain natural radioactive isotopes, mainly 226Ra from the uranium decay series and 228Ra from the thorium series. Approximately 40% of the total amount of radium remains underground as radioactive deposits, but 225 MBq of 226Ra and 400 MBq of 228Ra are released daily into the rivers along with the other mine effluents from all Polish coal mines. Technical measures such as inducing the precipitation of radium in gobs, decreasing the amount of meteoric inflow water into underground workings, etc. have been undertaken in several coal mines, and as a result of these measures, the total amount of radium released to the surface waters has diminished by about 60% during the last 5-6 years. Mine water can have a severe impact on the natural environment, mainly due to its salinity. However, associated high levels of radium concentration in river waters, bottom sediments and vegetation have also been observed. Sometimes radium concentrations in rivers exceed 0.7 kBq/m3, which is the permitted level for waste waters under Polish law. The extensive investigations described here were carried out for all coal mines and on this basis the total radium balance in the effluents has been calculated. Measurements in the vicinity of mine settling ponds and in rivers have given us an opportunity to study radium behaviour in river waters and to assess the degree of contamination. Solid waste materials with enhanced natural radioactivity have been produced in huge amounts in the power and coal industries in Poland. As a result of the combustion of coal in power plants, low-radioactive waste materials are produced, with 226Ra concentration seldom exceeding a few hundreds of Bq/kg. A different situation is observed in coal mines, where, as a result of precipitation of radium from radium-bearing waters, highly radioactive deposits are formed. Sometimes the radioactivity of such materials is extremely high; precipitates from coal mines may have radium concentrations of 400,000 Bq/kg--equivalent to 3% uranium ore. Usually, such deposition takes place underground, but sometimes co-precipitation of radium with barium takes place on the surface, in settling ponds and in rivers. Therefore management of solid waste with technologically enhanced natural radioactivity (TENR) is a very important subject. PMID- 11379078 TI - Radon exhalation from uranium mill tailings: experimental validation of a 1-D model. AB - TRACI, a model based on the physical mechanisms governing the migration of radon in unsaturated soils, has been developed to evaluate the radon flux density at the surface of uranium mill tailings. To check the validity of the TRACI model and the effectiveness of cover layers, an in situ study was launched in 1997 with the French uranium mining company, COGEMA. The study consisted of continuous measurements of moisture content, suction, radon concentration at various depths inside a UMT cover, and flux density at its surface. An initial analysis has shown that radon concentration and flux density, as calculated with a steady state diffusion model using monthly averaged moisture contents, are in good agreement with measured monthly averaged concentrations and flux densities. PMID- 11379079 TI - New perspectives in anaerobic digestion. AB - The IWA specialised group on anaerobic digestion (AD) is one of the oldest working groups of the former IAWQ organisation. Despite the fact that anaerobic technology dates back more than 100 years, the technology is still under development, adapting novel treatment systems to the modern requirements. In fact, most advances were achieved during the last three decades, when high-rate reactor systems were developed and a profound insight was obtained in the microbiology of the anaerobic communities. This insight led to a better understanding of anaerobic treatment and, subsequently, to a broader application potential. The present "state-of-the-art" paper, which has been written by members of the AD management committee, reflects the latest achievements and sets future lines for further development. PMID- 11379080 TI - Comparison of enumeration methods for ammonia-oxidizing bacteria. AB - MPN, antibody, fluorescent in situ hybridization (FISH) and dot blot hybridization methods for enumeration of ammonia-oxidizing bacteria were applied to various samples from suspended systems. As for FISH, reoptimization of an oligonucleotide probe Nso 190 was attempted. The advantages and disadvantages of these methods were discussed. MPN appeared to have disadvantages of underestimation. Dot blot hybridization had lower detection limit and higher reliability than other methods. It could be applied to mixed liquors and effluents of municipal wastewater treatment plant (WWTPs), lab-scale reactor and enrichment cultures of ammonia-oxidizing bacteria. FISH could be successfully applied to high ammonium loading system such as enrichment cultures, but weak signal, auto-fluorescence and non-specific binding could cause problems when applied to low ammonium loading systems such as effluents of WWTPs and river water. Mixed liquor of municipal WWTP seemed to be a critical case for FISH. PMID- 11379081 TI - Feed-forward control of the external carbon flow rate in an activated sludge process. AB - Biological nitrogen removal in an activated sludge process is obtained by two biological processes: nitrification and denitrification. Denitrifying bacteria need sufficient amounts of readily metabolized carbon. The objective of this paper is to develop an automatic control strategy for adjusting the external carbon flow rate so that the nitrate concentration in the last anoxic compartment is kept at a low pre-specified level. A simple model based feed-forward control combined with a standard feedback PI controller is suggested. Simulation results show that the suggested controller, despite being simple, effectively attenuates process disturbances. PMID- 11379082 TI - Stress protein expression in domestic activated sludge in response to xenobiotic shock loading. AB - Using the Western blot immunochemical analysis method, the heat shock protein, GroEL, was found to be either induced or repressed in activated sludge microorganisms exposed to a range of xenobiotics. At the EC25 concentration, pentachlorophenol (PCP), cadmium, nickel, 2,4-dichloroaniline, benzoquinone, 2,4 dinitrophenol, and 1,1,1-trichloroethane all rapidly induced measurable GroEL expression, even though the time-dependent response for each of these compounds was somewhat varied. Toluene and hydroquinone resulted in repression of GroEL expression to levels below that measured in the control mixed liquor. For PCP concentrations at or exceeding the EC25, there was a significant and consistent increase in effluent volatile suspended solids from activated sludge sequencing batch reactors relative to unstressed controls. These preliminary results indicate that stress proteins may serve as sensitive and rapid indicators of toxicity which can adversely impact treatment process performance in activated sludge systems. PMID- 11379083 TI - Effects of temperature on biodegradation characteristics of organic pollutants and microbial community in a solid phase aerobic bioreactor treating high strength organic wastewater. AB - The BOD removal rate and microbial community structure in a solid phase aerobic bioreactor using polyvinyl alcohol gel particles as packing material for the treatment of high strength organic wastewater were investigated at various temperatures. The BOD removal rate in the bioreactor increased when the temperature increased from 20 degrees C to 30 degrees C, 40 degrees C, and 50 degrees C, but it decreased when the temperature increased from 50 degrees C to 60 degrees C. Higher temperature enhanced the endogenous respiration of microbes in the bioreactor. The microbial community structure in the bioreactor was analyzed with quinone profile. The experimental results showed that the microbial community structure in the bioreactor was significantly affected by temperature. The dominant quinone of the microbes inhabiting the bioreactor was ubiquinone-8 at 30 degrees C, but that at 50 degrees C and 60 degrees C was menaquinone-7. It was estimated that the thermophilic Bacillus having menaquinone-7 dominated in the bioreactor at higher temperature. The microbial diversity in the bioreactor varied with temperature. PMID- 11379084 TI - Contribution of Pseudomonas spp. to phosphorus uptake in the anoxic zone of an anaerobic-anoxic-aerobic continuous activated sludge system. AB - A continuously operated laboratory-scale (32 L) nitrification denitrification biological excess phosphorus removal (NDBEPR) activated sludge system (modeled on the 3-stage Phoredox configuration) was maintained for 140 d. The transition from a non-biological excess phosphorus removal (BEPR) sludge to one exhibiting a strong BEPR mechanism was monitored. Mixed liquor seed inoculum was obtained from a full-scale single aerobic activated sludge installation and subjected to conditions conducive to BEPR, i.e. increasing influent acetate (HAc) concentrations. At a sludge age of 10 d with 100% HAc feed, the system was capable of removing a maximum of ca. 40 mgPO4-P/L from the bulk liquid; P/VSS of ca. 0.27 (mgP/mgVSS); and VSS/TSS of 0.53 (mgVSS/mgTSS) in the aerobic zone was attained. Although typical BEPR phosphorus transformation patterns were routinely observed, i.e. anaerobic phosphate release and aerobic phosphate uptake, phosphate uptake in the anoxic zone was also recorded indicating the presence of denitrifying phosphorus accumulating organisms (DPAOs) in the sludge community. The microbial community was screened (using both isolation and direct methods of analysis) for the presence of Pseudomonas spp. as this genus is known to perform both polyphosphate accumulation and denitrification processes. Isolation of anoxic mixed liquor bacteria on solid media and identification using the APl 20NE system resulted in the total dominance of the Pseudomonads (> 50%). However, direct fluorescent in situ hybridizations (FISH) revealed that Pseudomonas spp. only constituted ca. 3% of the total bacterial community indicating that other bacterial genera are contributing to simultaneous polyphosphate accumulation and denitrification processes in the anoxic zones of NDBEPR systems. PMID- 11379085 TI - Assessment of denitrification by the ordinary heterotrophic organisms in an NDBEPR activated sludge system. AB - The purpose of this study was to isolate and characterise the ordinary heterotrophic organisms (OHOs) present in a NDBEPR system in order to gain a better understanding of the organisms involved in denitrification as well as a more holistic and accurate evaluation of the OHO fraction attributable to denitrification in such a system. Heterotrophic bacteria were isolated from the pre- and secondary anoxic zones of the Darvill NDBEPR process and characterised according to their ability to reduce nitrates and/or nitrites under anoxic conditions. Results showed that the OHO fraction is more complex than currently accepted and, with respect to denitrification, can be more accurately subdivided into five functional groups, four of which interactively contribute to denitrification occurring in the system and one group that are non-denitrifying. These groups were defined as true denitrifiers (bacteria capable of both nitrate and nitrite reduction), incomplete denitrifiers (bacteria that reduced nitrates to nitrites with no further reduction of the nitrites produced), incomplete nitrite reducers (bacteria capable of both nitrate and nitrite reduction, however, exhibiting severe inhibition of nitrite reduction by nitrates), exclusive nitrite reducers (bacteria only capable of reducing nitrites) and non denitrifiers (bacteria not capable of nitrate or nitrite reduction). PMID- 11379086 TI - Impact of seeding with nitrifying bacteria on nitrification process efficiency. AB - Seeding of nitrifying bacteria into the activated sludge process was studied both theoretically and experimentally. A simple model was developed for prediction of the effects of seeding of nitrifying bacteria from a separate stage into the activated sludge process. The purpose of seeding is to improve the treatment results and the process stability as well as to decrease the volume requirements of the process. Pilot plant studies were carried out at the Uppsala municipal wastewater treatment plant in order to evaluate the effects of seeding. One line was supplied with supernatant from dewatering of digested sludge and the nitrification process gave an activated sludge with a high fraction of nitrifying bacteria, suitable for seeding. The other line was supplied with pre-precipitated wastewater and with the excess sludge from the line treating the supernatant. The experimental results showed that nitrification could be obtained at sludge ages that would otherwise preclude nitrification. Performance relationships for the system developed, based on laboratory and on-line measurements were studied and are presented. The studies show that seeding may decrease the necessary volume needs for a stable nitrification process and that the effects could be predicted by use of a simple model. PMID- 11379087 TI - Transformation of lipids in activated sludge. AB - Transformation of lipids in activated sludge treatment plants is of interest for two reasons: lipids contribute 30-40% of the chemical oxygen demand (COD) in wastewater, and they may stimulate the growth of filamentous microorganisms in nutrient removal activated sludge plants. The transformation of lipids was investigated under aerobic and anoxic conditions by measuring the oxygen and nitrate uptake rates (OUR and NUR). The maximal OUR and NUR of long-chain fatty acid was found to be at the same level as acetate indicating that long-chain fatty acid was as easily consumable. However, the adsorption of long-chain fatty acid to surfaces of sludge flocs made it difficult to determine initial uptake rates of long-chain fatty acids. It was not possible to describe the hydrolysis rate of triacylglyceride by OUR and NUR to long-chain fatty acids because the hydrolysis rate was very slow. For a better description of the processes involved in transformation of lipids, a conceptual model was suggested. The processes in the suggested model were the adsorption/desorption of both triacylglyceride, and long-chain fatty acid onto surfaces of sludge flocs, hydrolysis of triacylglyceride by lipases and the uptake of long-chain fatty acid by bacteria under various conditions. The model can be helpful to structure design and evaluation of activated sludge experiment with lipids. PMID- 11379088 TI - Microbial storage products, biomass density, and settling properties of enhanced biological phosphorus removal activated sludge. AB - The relationships between bacterial storage products, density, and settling characteristics were determined in a laboratory-scale sequencing batch reactor (SBR) enhanced biological phosphorus removal (EBPR) system. Both long-term and single anaerobic-aerobic cycle variations in these properties were studied. Increased polyphosphate (PP) content of the biomass during long-term operation resulted in improved sludge volume index (SVI) values. End-aerobic phase (after phosphate (P) uptake) SVI values were consistently lower than end-anaerobic phase (after P release) values. Neither filamentous nor slime bulking were evident by microscopic observations. Biomass density increased at a rate of 1.2 mg/L per each 1% increase in biomass P content. End-aerobic phase samples had an average 25% higher buoyant density than end-anaerobic phase samples; which was attributed to aerobic P uptake. Biomass density was negatively correlated with SVI values, and SVI values increased sharply at low biomass density. A mathematical model developed by Mas et al. (1985) was modified to predict total cell density based on literature values of PP, glycogen (GLY), and poly-b-hydroxybutyrate (PHB) densities. Model predictions were in good agreement with experimental results, although improved measurement of PP density is required to improve model predictions. PMID- 11379089 TI - Application of COD fractionation by a new combined technique: comparison of various wastewaters and sources of variability. AB - A new respirometric method for COD fractionation was applied to various wastewaters collected on French treatment plants. Great variations were observed especially in the readily biodegradable fraction (RBCOD) ranging from 1 to 16% of total COD. Variability of the results among the origin can be explained by the reactions occurring in the sewers. In one of the towns studied, the wastewater samples collected directly in the sewer show COD characteristics very different from the sewage which reaches the treatment plant. By analysing the same wastewater after aerobic and anaerobic storage, it was proved that RBCOD respectively decreased or increased significantly. Seasonal fluctuations were also quantified, showing that RBCOD fraction decreased from 75 mg/L to -15 mg/L during the summer whereas the variation of soluble fraction was less important. PMID- 11379090 TI - Mechanical disintegration of sewage sludge. AB - Mechanical disintegration can be used for an accelerated and improved anaerobic digestion of excess sludge. The hydrolysis is the limiting step of this process. Mechanical disintegration can be used to disrupt the cell walls and to cause the release of the organic material from the cells. Particle size analysis describes the size reduction but is not suitable for characterising the release of the organic material and the cell disruption. Two biochemical methods were developed for these phenomena. One of the parameters provides information about the disruption of micro-organisms, the other one gives information about the release of organic material. Different ultrasonic homogenizers, a high pressure homogenizer and stirred ball mills were used for disintegration experiments using various parameters. The influences of a mechanical disintegration on the particle size and of the energy intensity on the disintegration were investigated. Further investigations had to detect the influence of the solid content on the disintegration results. For sludge with a higher solid content better results in terms of energy consumption could be achieved. An optimum of the bead diameter and the stress intensity in stirred ball mills could be detected. A comparison of the results of different methods of sludge disintegration shows that the investigated ultrasonic homogenizers are inferior to a high pressure homogenizer and a stirred ball mill in terms of energy consumption. PMID- 11379091 TI - Sludge hydrolysate as a carbon source for denitrification. AB - External carbon sources such as methanol and acetate are applied in order to improve the denitrification efficiency in biological nitrogen removal processes. Excess sludge wasted from the treatment plants contains organic carbon that could be used as a feasible carbon source for denitrification. However, the excess sludge needs to be hydrolyzed first before being utilized as a carbon source. The objective of this research is to evaluate the biodegradability of the sludge hydrolysate obtained using alkaline, acid, autoclaved and combined solubilization methods. The experimental results suggest that the method of solubilization has an impact on the biodegradability of sludge hydrolysate. Autoclaved alkaline sludge hydrolysate gave the fastest denitrification rate (250 mgN/gMLSSCOD/d) due to fastest protein degradation (247 mgCOD/gLSSCOD/d) and other COD degradation rates (370 mgN/gMLSSCOD/d). Alkaline, autoclaved and acid autoclaved sludge hydrolysates gave nearly the same denitrification rates (185 to 200 mgN/gMLSSCOD/d). Protein degradation rate was the highest for autoclaved alkaline sludge hydrolysate (247 mgCOD/gMLSSCOD/d) whereas carbohydrate degradation rate was the fastest for autoclaved acid sludge hydrolysate (91 mgCOD/gMLSSCOD/d). For the same enzyme activity, different carbohydrate and protein degradation activities have been found suggesting the presence of easily and slowly biodegradable portions in them. PMID- 11379092 TI - Evaluation of nitrification-inhibition measurements. AB - A screening method for nitrification-inhibition determination has been evaluated at two laboratories, one in Sweden and one in Spain. Allyl-thiourea (ATU), methanol, chromium and zinc were used as reference toxicants in combination with different kinds of full-scale sludge and sludge produced in the laboratory. Different results were obtained with different combinations of activated sludge and toxicants. It was found that activated sludge often exposed to small amounts of a variety of toxic substances may build up a general resistance to toxic substances. Specific adaptation to certain substances was also observed. Domestic activated sludge responded in the same way to ATU regardless of whether it originated from Sweden or Spain. Synthetic sludge and domestic sludge exhibited the same inhibition of nitrification when tested with ATU as a toxicant. Synthetic activated sludge from two laboratory-scale plants run in parallel did not exhibit exactly the same sensitivity to certain toxicants. The variation in repeated tests with the synthetic sludges was about the same as for repeated tests with full-scale sludge and for other biological toxicity tests. Limits of detection were calculated to be about 5% for a single sample with three blanks as references at both the laboratories. PMID- 11379093 TI - Real-time control strategies for predenitrification-nitrification activated sludge plants biodegradation control. AB - This paper presents the real-time control strategies developed to regulate both the ammonia and nitrate concentration in the effluent of the new Vitoria WWTP (Spain). Nitrate control aims at the optimal use of the denitrification potential at any moment. For this purpose, the proposed control algorithm continuously adapts the internal recycle flow in order to maintain a desired nitrate set-point in the anoxic zone. Ammonia control aims at maintaining the required average concentration of ammonia in the effluent by manipulating the Dissolved Oxygen (DO) set-point. The control strategies have been based on a hierarchical structure where a high-level or supervisory control selects the set-point of the low-level or conventional controllers. The design of the controllers was carried out using the Quantitative Feedback Theory QFT for the design of robust control systems. Moving average values of some variables have been introduced in order to eliminate the perturbations associated with the daily 24-hour profiles. The controllers have been verified using long-time dynamic simulations based on a mathematical model previously calibrated in pilot plant. Influent load and temperature used in the simulations correspond to the real values measured in the full-scale WWTP during 12 months. The results obtained in the simulations show the good performance and stability of the control strategies independently from external disturbances. A short-time experimental verification of the controllers in pilot plant with real wastewater is also presented. PMID- 11379094 TI - Denitrification in submerged biofilters of concentrated-nitrate wastewater. AB - To understand the possibilities and limitations of the treatment of concentrated nitrate in wastewater, a space-saving biofilm process was introduced to investigate how efficiently concentrated nitrate was utilized in a biofilm process. Lab-scale submerged biofilters stuffed with plastic pall-ring media were developed for the treatment of wastewater containing high nitrate concentration. Two identical biofilters were operated at 2 and 4 hr media HRT in parallel and the concentration of nitrate in the influent was increased stepwise from 50 to 3000 mg-N/L. The concentration of NOx in the effluent was measured at various volumetric loading rates that were proportional to influent nitrate concentrations. There was a critical point for volumetric loading rate (9 kg NO3 N/m3/day) distinguishing the biofilter performances. At the volumetric loading rate below this critical point, the effluent NOx concentration was maintained steadily in spite of the drastic change in the loading rates. However, the effluent NOx concentration severely fluctuated for the small change when the loading rate was higher than the critical point. It was found that the reactor performance was closely related with the biomass concentration (g-dw/ring) in the biofilters. PMID- 11379095 TI - Wastewater treatment and nitrogen removal using submerged filter systems. AB - The performance of two submerged filter systems: a two filters-in-series system and a single combined filter system, in treating a strong nitrogenous wastewater with nitrogen concentration of 480 mg/L was evaluated. Both systems were equally effective in removing up to 90% of nitrogen and 98% of COD from the wastewater for loading rates up to 5 kg COD/m3.d and 0.5 kg N/m3.d. The second system in which anaerobic, anoxic, and aerobic zones were incorporated in a single filter offers a greater flexibility in treatment in that by repositioning the locations of the aeration point and effluent recycling inlet, the zonal volumes can be altered easily to treat wastewaters with different COD and nitrogen concentrations. PMID- 11379096 TI - Biological phosphorus and nitrogen removal in a sequencing batch moving bed biofilm reactor. AB - Biological phosphorus and nitrogen removal in biofilm processes have a potential advantage compared to activated sludge processes, because of less vulnerability with respect to sludge loss and because biofilm processes, in general, are more compact with a smaller footprint. Experiments have been carried out in a moving bed biofilm reactor operated as a sequencing batch reactor (SBR), with simultaneous nitrification, phosphorus uptake and denitrification in the aerobic phase. In order to achieve good phosphorus and nitrogen removal, the length of the anaerobic period should be tuned to achieve near complete removal of easily biodegradable COD in the anaerobic period, and the length of the aerobic period should be long enough for complete nitrification. The total COD-loading rate must be at the same time be kept high enough to achieve a net growth of biomass in the reactor. PMID- 11379097 TI - Feasibility of the BIOFIX-process for treatment of municipal wastewater. AB - The feasibility of the so-called BIOFIX-process to treat municipal wastewater was investigated in a pilot-plant. The innovative element of this process is that carrier material to which a biofilm is attached is recirculated between a sorption reactor to take up COD from the wastewater and a reactor where this COD is used for (post-) denitrification. In between the sorption and denitrification reactor the wastewater passes two subsequent reactors for removal of the remaining COD and for nitrification, respectively. The results showed that COD uptake by the biofilm in the sorption reactor with a maximum of 34% of the influent load was far below expectations and did not meet the COD requirement for denitrification (50-60% uptake). Also, 9-21% of the influent load of ammonia was taken up by the biofilm. In the denitrification reactor this ammonia was released to the bulk and in this manner discharged with the effluent without having passed the nitrification reactor. Nitrification was inhibited by the presence of high concentrations of suspended solids (50-60 mgl-1) discharged from a COD removal reactor. Together these bottle-necks caused effluent concentrations which were well above the effluent demands and it therefore can be concluded that the BIOFIX process is not a feasible process to treat domestic wastewater. PMID- 11379098 TI - External nitrification in biological nutrient removal activated sludge systems. AB - A biological nutrient removal (BNR) activated sludge (AS) scheme incorporating external nitrification in a fixed media system is evaluated. A laboratory scale investigation of the scheme indicates that it holds considerable potential for BNRAS system intensification through major reduction in sludge age and oxygen demand and significant improvement in sludge settleability. Because the BNRAS system is not required to nitrify, its anoxic mass fraction can be considerably enlarged at the expense of the aerobic mass fraction creating conditions that (i) allow it to achieve high N removals with domestic wastewaters with high TKN/COD ratios and (ii) promote anoxic P uptake polyphosphate accumulating organisms (PAO) to develop in the system. From this, and earlier investigations with conventional BNR systems, it appears that anoxic P uptake biological excess P removal (BEPR) is only about two thirds of aerobic P uptake BEPR. Inclusion of anoxic P uptake PAOs in, and exclusion of nitrifiers from, the BNRAS system are not essential for the scheme. However, conditions that promote aerobic P uptake to maximize BEPR, are also conducive to nitrifier growth, which, if supported in the BNRAS system, would require virtual complete nitrification in the fixed media system to avoid nitrate interference with BEPR. Before the scheme can be implemented at large scale, an engineering and economic evaluation is required to quantify its potential benefits and savings. PMID- 11379099 TI - Development and verification of design and operation criteria for the step feed process with nitrogen removal. AB - The step feed process with three stages of denitrification-nitrification reactors has been studied using simulations in order to develop criteria for the optimum selection of the design parameters and to propose efficient operational strategies. To verify the simulation results experimental studies in a pilot plant of 1100 litres were carried out. The simulation studies showed that the optimum influent flow distribution to the three anoxic reactors is in the range of 40-40-20% and 33-33-34% depending on the wastewater characteristics and effluent requirements. These two latter conditions and in turn the influent flow distribution determine the anoxic and aerobic reactor volumes. The reduction of the dissolved oxygen in the two first aerobic reactors and the use of facultative zones in the final D-N stage are proposed as operational strategy. The experimental results proved the validity of the criteria developed for design and operation. A high capacity of the IAWQ activated sludge model No. 1 to predict the performance of the step feed process was observed. PMID- 11379100 TI - Simultaneous nitrification and denitrification in biofilters with real time aeration control. AB - The aim of this article is to present a new biological aerated filter (BAF) for nitrogen removal based on simultaneous nitrification and denitrification. Contrary to the systems which integrate both an aerated and a non-aerated zone to allow complete nitrogen removal in one compact or two different units (pre denitrification and nitrification), this upflow BAF system is based on the principle of simultaneous nitrification and denitrification since the filter is completely aerated. The denitrification process is possible due to the diffusion effect which dominates biofilm processes. The real time aeration control allows us to maintain a low dissolved oxygen value (0.5 to 3 mg O2/l). In this case, the biofilm will not be fully (or less) penetrated with oxygen and denitrification will be carried out in a large part of the biofilm. Therefore, nitrification and denitrification is running simultaneously in different depths of the biofilm. By using 50% less air this BAF gave the same results (less than 20 mg TN/l) on pilot plant as a classical nitrification and denitrification BAF (Toettrup et al., 1994). Less recirculation was necessary to achieve the same denitrification. PMID- 11379101 TI - Performance characterization of anaerobic sequencing batch reactor process for digestion of night soil. AB - Laboratory experiments were conducted to investigate the performance of an anaerobic sequencing batch reactor (ASBR) process for night soil treatment. Performances of the reactors were evaluated at an equivalent hydraulic retention time (HRT) of 10 days with an equivalent loading rate of 2.6 kgVS/m3/d (3.1 kgCOD/m3/day) at 35 degrees C. Digestion of a night soil was possible using the ASBR at an HRT of 10 days in spite of high concentration of ammonia nitrogen and settleable solids. Solids were accumulated rapidly in the ASBRs, and their concentrations were 2.3-2.4 times higher than that in a completely mixed control reactor. Remarkable increases in gas production were observed in the ASBRs compared with the control reactor. Average increases in equivalent daily gas production from the ASBRs were 205-220% compared with that from the control run. The ASBR with reaction period/thickening period ratio (R/T ratio) of 1 showed a little higher gas production and organic removal efficiency than that with R/T ratio of 3. Volatile solids removals based on supernatant of the ASBRs were 12 14% higher than that of the control reactor. Thus, the ASBR was a stable and effective process for the treatment of night soil having high concentration of settleable organics and ammonia nitrogen. PMID- 11379102 TI - Algae-removal performance of a fluidized-bed biofilm reactor system for lake water treatment. AB - The algae removal efficiency of a pilot plant--based on a fluidized-bed biofilm reactor system for treating--was investigated. This system does not require back washing because the fluidized-bed suffers no clogging. Moreover, the system uses dissolved oxygen in the influent water for aerobic biological treatment without the need for additional aeration equipment. This, it is an easy-maintenance, low energy system for purifying eutrophic lake water. The system was operated continuously at a flow rate of 1500 m3/d for nine months at Tsuchiura Port in Lake Kasumigaura. And concentrations of chlorophyll-a and dissolved oxygen in both the influent and effluent water were continuously monitored. In summer (August to September) when water bloom occurred, the average efficiency of chlorophyll-a removal was 64% at an average influent chlorophyll-a concentration of 137.8 micrograms/L. Over the entire experimental period of nine months, the average daily amount of removed chlorophyll-a was 40.3 g/d at an average influent chlorophyll-a concentration of 89.5 micrograms/L. By analyzing the relationship between the amount of removed chlorophyll-a and the consumption of dissolved oxygen, it was estimated that almost all of the algae trapped in the reactor was biologically degraded. PMID- 11379103 TI - Biofilm structure and mass transfer in a gas phase trickle-bed biofilter. AB - Mass transport phenomena occurring in the biofilms of gas phase trickle-bed biofilters are investigated in this study. The effect of biofilm structure on mass transfer mechanisms is examined using experimental observation from the operating of biofilters, microelectrode techniques and microscopic examination. Since the biofilms of biofilters used for waste gas treatment are not completely saturated with water, there is not a distinguishable liquid layer outside the biofilm. Results suggest that due to this characteristic, gas phase substrates (such as oxygen or volatile organic compounds) may not be limited by the aqueous phase because transport of the compound into the biofilm can occur directly through non-wetted areas. On the other hand, for substrates that are present only in the liquid phase, such as nitrate, the mass transfer limitation is more serious because of the limited liquid supply. Microscopic observations show that a layered structure with void spaces exists within the biofilm. Oxygen concentration distributions along the depth of the biofilms are examined using an oxygen microelectrode. Results indicate that there are some high dissolved oxygen zones inside the biofilm, which suggests the existence of passages for oxygen transfer into the deeper sections of the biofilm in a gas phase trickle-bed biofilter. Both the low gas-liquid mass transfer resistance and the resulting internal structure contribute to the high oxygen penetration within the biofilms in gas phase trickle-bed biofilters. PMID- 11379104 TI - Simplified and Monod kinetics in one-dimensional biofilm reactor modelling: a comparison. AB - A theoretical study supported by some experimental tests has been carried out with the aim of comparing one-dimensional (1-D) biofilm reactor models that use simplified (zero- and first-order) and Monod kinetics. Two different situations have been compared: one rate-limiting substrate with or without liquid film diffusion. The results obtained show that the use of a simplified kinetic approach compared to the Monod kinetic approach determines (1) an unjustified overestimate of the removal rate, especially for thin biofilms, and (2) an excessive overestimate of the liquid film layer thickness necessary to justify high kinetic orders. Even if recent research projects show that biofilm structure is more complicated than the one assumed in the modelling approach used in this study, nevertheless 1-D models still now continue to be the only ones that can reasonably support process engineers in biofilm reactor design, due to their intrinsic simplicity and the need for small sets of input data and parameters that can be obtained theoretically or often empirically. PMID- 11379105 TI - Contributions of biofilm versus suspended bacteria in an aerobic circulating-bed biofilm reactor. AB - This study demonstrated that, during the two-step biodegradation of toluene in an aerobic circulating-bed biofilm reactor, biofilm and suspended bacteria played critical roles. Although the suspended bacteria were less than 1% of the total amount of biomass in the system, they transformed up to 30% of the toluene into its intermediate in the bulk liquid phase. On the other hand, most of the toluene intermediate was removed inside the biofilm, where diffusion resistance reduced the toluene concentration, thereby relieving inhibition to the degradation reaction of the intermediate. The suspended bacteria are most important for rapidly biodegraded substrates, for which diffusion limitation controls the kinetics in the biofilm. They lose importance when the effects of an inhibiting substrate must be overcome. PMID- 11379106 TI - Single stage biological nitrogen removal by nitritation and anaerobic ammonium oxidation in biofilm systems. AB - In full scale wastewater treatment plants with at times considerable deficits in the nitrogen balances, it could hitherto not be sufficiently explained which reactions are the cause of the nitrogen losses and which micro-organisms participate in the process. The single stage conversion of ammonium into gaseous end-products--which is henceforth referred to as deammonification--occurs particularly frequently in biofilm systems. In the meantime, one has succeeded to establish the deammonification processes in a continuous flow moving-bed pilot plant. In batch tests with the biofilm covered carriers, it was possible for the first time to examine the nitrogen conversion at the intact biofilm. Depending on the dissolved oxygen (DO) concentration, two autotrophic nitrogen converting reactions in the biofilm could be proven: one nitritation process under aerobic conditions and one anaerobic ammonium oxidation. With the anaerobic ammonium oxidation, ammonium as electron donor was converted with nitrite as electron acceptor. The end-product of this reaction was N2. Ammonium and nitrite did react in a stoichiometrical ratio of 1:1.37, a ratio which has in the very same dimension been described for the ANAMMOX-process (1:1.31 +/- 0.06). Via the oxygen concentration in the surrounding medium, it was possible to control the ratio of nitritation and anaerobic ammonium oxidation in the nitrogen conversion of the biofilm. Both processes were evenly balanced at a DO concentration of 0.7 mg/l, so that it was possible to achieve a direct, almost complete elimination of ammonium without addition of nitrite. One part of the provided ammonium did participate in the nitritation, the other in the anaerobic ammonium oxidation. Through the aerobic ammonium oxidation into nitrite within the outer oxygen supplied layers of the biofilm, the reaction partner was produced for the anaerobic ammonium oxidation within the inner layers of the biofilm. PMID- 11379107 TI - Removal of organics from wastewater using a novel biological hybrid system. AB - This paper summarizes the results obtained using the hybrid aerated submerged fixed-film (HASFF) process. HASFF is an innovative system comprising a four compartment reactor having an array of fixed ceramic plates maintained under diffused aeration to support attached biomass, with activated sludge recycle to promote suspended growth in the reactor. Wastewater from a municipal treatment plant was fed to the reactor and its activated sludge was used for recycling in the hybrid system. Four pilot units were operated in the plant at various hydraulic retention times, HRTs (2, 4, 6 and 8 h), using primary-settled wastewater under organic loading rates up to 0.7 g BOD/gMLTVS. d. Data obtained showed that the overall BOD percentage removal efficiencies were consistently above 94.0% at all HRTs including the 2 hours while the COD percentage removal efficiencies ranged between 65.7-76%. The effluent's mean filtered BOD concentration ranged between 4.5-7.5 mg/l whereas the mean filtered COD concentration ranged between 70.0-89.6 mg/l. Increasing the hydraulic loading rate by four-folds from 0.08 to 0.32 m3/m2.d had a minor effect on the unit's BOD and COD percentage removal efficiencies indicating a robust biological process that is resilient to hydraulic shock loads, thereby offering a viable upgrading option. PMID- 11379108 TI - Advanced treatment of sewage by pre-coagulation and biofilm process. AB - Treatment performance and operational parameters of a pre-coagulation and biofilm process were experimentally discussed with the pilot-scale plant treating actual municipal sewage. Perfect nitrification was accomplished within total hydraulic retention time (HRT) of 4 h at 20% volumetric added ratio of attached-growth media of biofilm in the aerobic tanks under temperature conditions as low as 17 degrees C. Treatment efficiencies in organic matter, T-N and T-P were high and stable with the total HRT of 8 h in the biological reactor. Concentrations of T-N and T-P in effluent were kept about 2 mgN/L and under 1 mgP/L, respectively. Attached growth bacterial mass and the potential activity of nitrification and denitrification in each tank in cold season were high enough to accomplish excellent treatment performance. Then the process was proved to be applicable to municipal sewage treatment for removal of nitrogen and phosphorus as well as organic matter. PMID- 11379109 TI - Stability of a lab-scale biofilm for simultaneous removal of phosphorus and nitrate. AB - A lab-scale biofilm reactor for simultaneous removal of phosphorus and nitrate was operated for one and a half years. Despite using only well defined synthetic wastewater and well defined operation, the activity varied significantly over the months. It was speculated that microbial population shifts were causing this phenomenon. This could also explain a sudden break down of the process following a slight change in the operation. Over shorter periods of time (time-scale: days), the biofilm could be considered stable enough to perform series of comparable batch experiments. Batch experiments with different start concentrations of acetate, nitrate or phosphate were conducted. These verified 0.5 and 0 order removal rates in the bulk water depending on the concentration. This was taken as an indication of a zonation of the biofilm. Due to the measured variability in the activity and due to the importance of the history of the bacteria when considering biological P removal, on-line measurements are strongly recommended for research on this subject. Microbial characterisation methods are recommended as an assisting tool in further research. PMID- 11379110 TI - Rapid-rate thermophilic, single-stage and two-phase methane fermentation of synfuel-industry wastewaters. AB - Thermophilic (55 degrees C) anaerobic digestion of wastewaters from wet carbonization of Minnesota peat was conducted in a simple, upflow digester at an HRT of one day to provide a methane yield of 0.20 SCM/kg VS added and a BOD5 reduction of 85%. Carbonization of Maine peat conducted for a much longer duration of thermal treatment produced recalcitrant and potentially toxic end products that reduced the biodegradability of wastewaters. Methane fermentation of the Maine-peat wet-carbonization waste was inhibited at an HRT of one day in the single-stage digester. However, two-phase digestion with separate acid-phase fermentation to promote hydrolytic degradation of toxic end products followed by separate methane fermentation at an HRT of 2 days exhibited a methane yield of 0.21 SCM/kg VS added and a BOD5 reduction of 76%. Volatile fatty acids profiles along the depths digesting cultures indicated that single-stage and two-phase digestion could be accomplished at one-half the reactor residence times (one day for single-stage and three days for two-phase) used in this research. PMID- 11379111 TI - Hybrid modelling of anaerobic wastewater treatment processes. AB - This paper presents a hybrid approach for the modelling of an anaerobic digestion process. The hybrid model combines a feed-forward network, describing the bacterial kinetics, and the a priori knowledge based on the mass balances of the process components. We have considered an architecture which incorporates the neural network as a static model of unmeasured process parameters (kinetic growth rate) and an integrator for the dynamic representation of the process using a set of dynamic differential equations. The paper contains a description of the neural network component training procedure. The performance of this approach is illustrated with experimental data. PMID- 11379112 TI - A molecular method to study population and activity dynamics in anaerobic digestors. AB - The applicability of a new molecular fingerprinting method (Single Strand Conformation Polymorphism) to study the microbial populations of anaerobic digestors was investigated. After extraction of total nucleic acids, the 16S rDNA and 16S rRNA molecules were amplified and the amplicons were separated by SSCP electrophoresis. Characteristic and complex peak patterns were obtained, where each peak could be correlated with the 16S rDNA sequence of one micro-organism. The rDNA peak patterns should consist of the most abundant sequences and thus would reflect the diversity of prominent species of different digestors. Ribosomal DNA patterns were compared to rRNA patterns and revealed the bacteria that were the most active metabolically. The SSCP method also revealed dynamic changes in the presence and activity of populations, following perturbations such as an acidic shock which caused an increase in activity of two species. After cloning the 16S rDNA, the species corresponding to the peaks of interest, such as the archaeal species, could be identified by screening the clones according to their SSCP patterns and sequencing the 16S rDNA. PMID- 11379113 TI - Population dynamics of anaerobic microbial consortia in thermophilic granular sludge in response to feed composition change. AB - A thermophilic UASB reactor was operated at 55 degrees C for greater than 470 days in order to investigate the effects of feed composition on the changes in microbial community structure where thermophilic granular sludge was used as the inoculum source. The feed compositions were changed with cultivation days; phase 1 (1-70 days), alcohol distillery wastewater; phase 2 (71-281 days), artificial acetate wastewater; phase 3 (282-474 days), artificial sucrose wastewater. During the first one month of each phase, the methanogenic activity and cell density of methanogens quantified by fluorescence in situ hybridization (FISH) drastically changed as a result of shift in feed composition. When artificial acetate wastewater was used as feed, retained granular sludge was partially disintegrated due to a decrease in the number of symbiotic bacterial community members: acetogens (acidogens) and hydrogenotrophic methanogens. In contrast, when the feed was shifted to sucrose (phase 3), granulation of biomass was promoted by a remarkable proliferation of the symbiotic community. The presence of hydrogen utilizing methanogens and acetogens (acidogens) are shown to be effective for the enhancement of thermophilic granulation. The cell density of methanogens determined by FISH was strongly correlated with the methane-producing potential of the retained thermophilic granular sludge. PMID- 11379114 TI - Anaerobic degradation of 2,4,6-trinitrotoluene in granular activated carbon fluidized bed and batch reactors. AB - In this study, an anaerobic fluidized bed reactor (AFBR) was used to treat a synthetically produced pink water waste stream containing trinitrotoluene (TNT). The synthesized waste consisted of 95 mg/l-TNT, the main contaminant in pink water, which was to be co-metabolized with 560-mg/l ethanol. Granular activated carbon was used as the attachment medium for biological growth. TNT was reduced to a variety of compounds, mainly 2,4,6-triaminotoluene (2,4,6-TAT), 2,4-diamino 6-nitrotoluene (2,4-DA-6-NT), 2,6-diamino-4-nitrotoluene (2,6-DA-4-NT), 2-amino 4,6-dinitrotoluene (2-A-4,6-DNT), and 4-amino-2,6-dinitrotoluene (4-A-2,6-DNT). These conversions resulted through the oxidation of ethanol to carbon dioxide under anoxic conditions, or reduction to methane under methanogenic conditions. The anaerobic reactor was charged with 1.0 kg of 16 x 20 U.S. Mesh Granular Activated Carbon (GAC) and was pre-loaded with 200 g of TNT prior to the addition of the mixed seed culture. During the first three weeks of operation, ethanol was completely degraded and no methane was produced. Effluent inorganic carbon revealed stoichiometric conversion of the feed ethanol to dissolved inorganic carbon with accumulation of carbon dioxide in the headspace of the reactor. GAC extraction showed incremental reduction of the nitro groups to amino groups, with 2,4,6-TAT as the final product. After three weeks, the oxygen from the nitro groups was depleted and methane production commenced. The reproducibility of this phenomenon was confirmed by repeating the experiment in the same manner using an identical AFBR. Furthermore, serum bottle tests were conducted using TNT loading ratios of 0.2, 0.4, 0.8, 1.0 g-TNT/g-GAC as well as experiments in the absence of GAC. Similar behavior to that of the columns was observed, with degradation rates varying according to the particular condition. GAC greatly enhanced the degradation rates and the higher TNT loading resulted in slower degradation rates of ethanol. PMID- 11379115 TI - Study of microbial community of brewery-treating granular sludge by denaturing gradient gel electrophoresis of 16S rRNA gene. AB - The microbial community structure of granular sludge from an upflow anaerobic sludge blanket (UASB) reactor treating brewery effluent was studied by denaturing gradient gel electrophoresis (DGGE). Twelve major bands were observed in the DGGE fingerprint for the Bacteria domain and four bands for the Archaea domain. Of the bacterial bands observed, six were successfully purified and sequenced. Among them, three were related to the gram-positive low G + C group, one to the Delta subclass of the Proteobacteria, one to the Gamma subclass, and one to the Cytophaga group with no close related sequence. The 16S rRNA sequences of the four archaeal bands were closely associated with Methanosaeta concilii and Methanobacterium formicum. PMID- 11379116 TI - Biolog for the determination of microbial diversity in activated sludge systems. AB - In this study, different carbon source profiles were generated by inoculating Biolog GN microwell plates, with different dilutions of microbial communities from a number of activated sludge systems. This led to the successful generation of patterns reflecting diversity and evenness in the different systems. The high number of substrates utilized at the lower dilutions (10(-1) and 10(-2)) indicated a high microbial diversity in the community, but not necessarily evenness of each species. Evenness of each species was reflected upon further dilution. Our results indicated differences in the microbial community composition amongst some of the activated sludge systems studied. These differences were not specifically related to phosphate removing and non phosphate removing systems. PMID- 11379117 TI - 16S rRNA in situ probing for the determination of the family level community structure implicated in enhanced biological nutrient removal. AB - Knowledge of a discrete physiological group capable of excess biological phosphate removal (EBPR) remains unclear. Consequently, microbial community analysis of an enhanced continuous laboratory-scale activated sludge process displaying a strong EBPR mechanism was conducted. Unit design was configured upon the three-stage Phoredox process and characterization of the activated sludge bacterial community was carried out using fluorescent in situ hybridization (FISH) techniques. Fixed activated sludge samples were hybridized with fluorescently labeled oligonucleotide probes targeting the following bacterial phylogenetic divisions: a kingdom level probe specific for all bacteria (EUB338); family level probes specific for the alpha, beta and gamma subclasses of the class Proteobacteria; Gram positive bacteria with a high (G + C) DNA content (GPBHGC) or Actinobacteria; the Cytophaga-Flavobacterium (CF) subclass within the Cytophaga-Flavobacterium-Bacteriodes division; and genus level probes specific for Pseudomonas spp., Aeromonas spp., and Acinetobacter spp. Bacterial predominance between the anaerobic, anoxic and aerobic zones of the EBPR sludge were comparable and appeared as follows; beta (22%), alpha (19%), gamma (17%), GPBHGC (11%) and CF (8%). The incidence of Acinetobacter spp. appeared to be generally low with counts amounting to < 9% of the total bacterial count. A population shift in the alpha Proteobacteria subclass was evident between the non nutrient removal seed sludge and the EBPR sludge, implicating this group in EBPR. The overall results indicate that the beta and alpha Proteobacteria can be metabolically functional in EBPR processes and reiterate the functional misconception of Acinetobacter spp. in EBPR systems. PMID- 11379118 TI - Characterization of microbial community in an activated sludge process treating domestic wastewater using quinone profiles. AB - The dynamics of microbial community structure of activated sludges in a small scale domestic wastewater treatment process were examined using a novel approach of quinone profiles. The composition and content of quinones in the activated sludges were analyzed monthly over a period of one year. More than 4 types of ubiquinones and 12 types of menaquinones were observed in the activated sludges, with the dominant quinones being ubiquinone (UQ)-8, menaquinone (MK)-7, followed by UQ-10, MK-8 and MK-6. The total quinone contents in the activated sludges varied from 0.93 to 2.68 mumol per gram of particle organic carbon. The molar ratio of ubiquinones to menaquinones (UK/MK) changed from 0.38 to 0.98, indicating that anaerobic bacteria dominated the microbial community of the activated sludges examined. The ratio of UQ/MK varied similar to that of dissolved oxygen in the bulk. The microbial diversity of the activated sludges calculated from the quinone compositions was 13.4-16.8. The diversity of menaquinones was much higher than that of ubiquinones, and increased slightly with increasing temperature. The microorganisms containing menaquinones appear to be sensitive to the change in temperature than those containing ubiquinones. PMID- 11379119 TI - River Water Quality Model no. 1 (RWQM1): I. Modelling approach. AB - Successful river water quality modelling requires the specification of an appropriate model structure and process formulation. Both must be related to the compartment structure of running water ecosystems including their longitudinal, vertical, and lateral zonation patterns. Furthermore, the temporal variability of abiotic boundary conditions may be important and must be incorporated by an appropriate choice of model parameters. A six-step decision procedure is proposed to achieve these objectives. The steps address the determination of the following model features: (1) temporal representation (dynamic or steady-state); (2) model dimensionality; (3) mixing; (4) advection; (5) reaction terms; and (6) boundary conditions. Numerical criteria based on process time constants and length scales provide a basis for these decisions. PMID- 11379120 TI - The effects of particle size distribution on the settleability of CSOs pollutants. AB - Over the past decades, flocculation and/or sedimentation processes have been adopted to remove pollutants from CSOs. It has been learned that major factors affecting settlement of pollutants are the particle size distribution, their settling velocities and their specific gravity. It is, therefore, a good idea to analyze the particle size distribution and settleability of CSOs pollutants in order to develop details in designing a process. Discussed in this study are pollutant characteristics of CSOs such as particle size distribution and settleability of pollutants. The power law function is applied and is found to be an effective and reliable index for expressing the particle size distribution of pollutants in CSOs. Based on the particle size spectrum analysis, the tendency toward settling and simultaneous flocculation-settling phenomenon of CSOs pollutants is described. Based on the regression analysis it is observed that the derived constants of curves representing settling velocity profile are proportional to the initial concentration of particles and to the beta-values of power law distributions. It is also revealed that the simultaneous flocculation settling processes are effectively described by the changes of the average particle diameter and of the beta-values of power law distributions. PMID- 11379121 TI - River Water Quality Model no. 1 (RWQM1): II. Biochemical process equations. AB - In this paper, biochemical process equations are presented as a basis for water quality modelling in rivers under aerobic and anoxic conditions. These equations are not new, but they summarise parts of the development over the past 75 years. The primary goals of the presentation are to stimulate communication among modellers and field-oriented researchers of river water quality and of wastewater treatment, to facilitate practical application of river water quality modelling, and to encourage the use of elemental mass balances for the derivation of stoichiometric coefficients of biochemical transformation processes. This paper is part of a series of three papers. In the first paper, the general modelling approach is described; in the present paper, the biochemical process equations of a complex model are presented; and in the third paper, recommendations are given for the selection of a reasonable submodel for a specific application. PMID- 11379122 TI - Small diameter gravity sewers: self-cleansing conditions and aspects of wastewater quality. AB - The construction of conventional sewerage systems in small communities, with pipes laid on a uniform slope and manholes regularly spaced, is sometimes not economically feasible, because of the high costs of sewer installation. Under those circumstances, the small diameter gravity sewers (SDGS) have often proven to be substantially less costly than conventional sewers. Typically, in SDGS systems the wastewater from one or more households is discharged into an interceptor tank (or a single compartment septic tank). The settled effluent is discharged afterwards into small diameter sewers operating under gravity. In this paper, special emphasis is given to the analysis of self-cleansing conditions and to the analysis of risks of sulphide generation and occurrence of septic conditions in SDGS systems. For the evaluation of the self-cleansing conditions, the critical velocity and the critical shear stress were computed according to the Shields equation. The forecasting of dissolved oxygen concentrations and sulphide build-up along the lines, for different flow conditions, was done running an established wastewater quality model. PMID- 11379123 TI - The purification performance of infiltration basins fitted with pretreatment facilities: a case study. AB - In the south-east of France, the evacuation of stormwater by infiltration into the ground is being developed for large aquifer systems such as the ground water in the Rhone valley and in the eastern part of Lyons. A study proposal has been presented to the water management department of the conurbation of Lyons, aimed at quantifying, within a reasonably short space of time, the effects, in terms of transport of pollutants, of the stormwater infiltration system in the underground water in eastern Lyons. To this end, a one year duration experiment was carried out on the Venissieux infiltration basin which drains stormwater from a 380 hectares industrial catchment area. Its peculiar configuration also made it possible to acquire new knowledge on the qualitative operation of a few pretreatment facilities. After describing the operation of the basin and the experimental protocol, we shall present a body of data that we monitored and our conclusions about the behaviour of the pollution throughout the facilities. Then, we present methods used to assess the pollution removal performance of the infiltration basin and its pretreatment devices, the results obtained, and our conclusions about the impact of the infiltration basin on groundwater and soil. PMID- 11379124 TI - Sulfide production and wastewater quality--investigations in a pilot plant pressure sewer. AB - The relationship linking sulfide production rate and wastewater quality in terms of its biodegradability was studied using a pilot plant pressure sewer (inner diameter: 102 mm, length: 47 m). Furthermore, anaerobic transformations of wastewater organic matter were investigated. Wastewater characterization based on oxygen utilization rate (OUR) measurements and VFA analyses was employed. As wastewater quality parameters essential for the sulfide production, COD components and dissolved carbohydrate were focused on. Readily biodegradable substrate and fermentable, readily biodegradable substrate were better parameters than traditional dissolved COD for the prediction of sulfide production rates in a pressure sewer. From the results obtained, it was possible to integrate the sulfide production process with the transformation processes of wastewater organic matter in pressure sewers. PMID- 11379125 TI - New challenges for the management of plant nutrients and pathogens in the Waikato River, New Zealand. AB - The water quality of the Waikato River is currently much better than it was in the 1950s. Major improvements in the treatment of the sewage and industrial wastewaters which are discharged to the river mean that levels of indicator bacteria in the lower reaches of the river are now many times lower than in the past. Eve so, conditions are still not suitable for swimming, and blue-green algal blooms occur at times. Non-point or diffuse sources of contaminants now dominate the nutrient and pathogens budgets. Progressively-intensifying farming, particularly in lowland areas, is thought to contribute the majority of the contaminants found in the river. Future improvements in water quality will therefore depend more on activities like changes to farming practice--such as retiring the riparian margins of lowland tributaries of the river--than on further advances in wastewater treatment. PMID- 11379126 TI - Water resources protection today: end-of-pipe technology and cleaner production. Case study of the Czech Odra River watershed. AB - This paper reports on integrated watershed-based protection and sustainable use of water resources to increase the effectiveness of water pollution abatement. The approach includes improvements in end-of-pipe waste-water treatment technologies and implementation of Cleaner Production (CP) principles and policies within the watershed. An example of the general effectiveness of this approach is illustrated by the Czech Odra River Cleaner Production Project where reductions in pollution were achieved with improved industrial production. The CP theme is worth considering as an important challenge for the IWA. PMID- 11379127 TI - Dynamic phosphorus mass balance modeling of large watersheds: long-term implications of management strategies. AB - The principles of mass balance, compartment-flux diagramming, and dynamic simulation modeling are integrated to create computer models that estimate phosphorus (P) export from large-scale watersheds over long-term futures. These Watershed Ecosystem Nutrient Dynamics (WEND) models are applied to a 275,000 ha dairy-documented watershed and a 77,000 ha poultry-dominated watershed in northeastern USA. Model predictions of present-day P export loads are consistent with monitoring data and estimates made using P export coefficients. For both watersheds P import exceeds P export and P is accumulating in the agricultural soils. Agricultural and urban activities are major contributors to P export from both watersheds. Continued urban growth will increase P export over time unless wastewater management is substantially enhanced and/or rates of urban growth are controlled. Agriculture cannot rely solely on the implementation of increasingly stringent conservation practices to reduce long-term P export but must consider options that promote P input/output balance. The WEND modeling process is a powerful tool to integrate the diversity of activities in watersheds into a holistic framework. Model outputs are suited to assist managers to explore long term effects of overall watershed management strategies on P export in comparison to environmental and economic goals. PMID- 11379128 TI - Effects of aquatic macrophytes on physico-chemical conditions of three contrasting lowland streams: a consequence of diffuse pollution from agriculture? AB - Three lowland streams in developed pasture catchments with different farming intensities exhibited contrasting summer diurnal variations in pH, DO and temperature. These are ascribed to differences in dominant aquatic vegetation and their respective effects on shade, and on photosynthetic production and respiration within each stream. The stream dominated by submerged macrophytes had the greatest amplitude swings in DO and pH, and DO levels of 86-128% saturation. Floating marginal macrophytes reduced photosynthetic inputs while providing additional organic loading for respiration, with consequent flat DO and pH curves and conditions not conducive to healthy stream ecosystems. The third stream was shaded by riparian plants, which inhibited photosynthetic effects on DO and pH so that diurnal variation was intermediate between the other two streams. The interaction between nutrients and increased insolation in agricultural catchments, in stimulating aquatic plants, needs to be better understood for managing the sustainability of stream habitats and ecosystems. PMID- 11379129 TI - Control of non-point source pollution by a natural wetland. AB - Wetland creation and restoration is a reliable and efficient technology for the remediation of contaminated water. Knowledge from the natural wetland systems would be necessary to enhance the operational efficiency of constructed wetlands. In this study, a mountainous wetland located in McDowell County, North Carolina, USA was selected to demonstrate the effects of the natural filtration and restoration system on the maintenance of surface water quality. The hydraulic retention time (HRT) for the wetland was 10.5 days based on the results from a dye release study. Water quality monitoring of the wetland was conducted from May to August 1997. One major storm event and baseline water quality samples were collected and analyzed. Analytical results indicate that this wetland removed a significant amount of non-point source (NPS) pollutants [more than 80% N removal, 91% of total suspended solid removal, 59% of total phosphorus removal, and 66% of chemical oxygen demand (COD) removal] caused by the studied storm event. Sediment accretion monitoring results indicate that the accretion rate in the wetland was only 4 mm/year. Therefore, the wetland would require 100 years to fill at the measured sediment accretion rate. The high organic content of sediments (16%) indicates that the wetland is building the characteristic organic layer on the bottom of the wetland. Results from this study would be very useful in the maintenance of natural wetlands and design of constructed wetlands for water treatment. PMID- 11379130 TI - Water quality response to riparian restoration in an agricultural watershed in Vermont, USA. AB - Achievement of management goals for Lake Champlain (Vermont/New York, USA and Quebec, Canada) will require reduction of agricultural phosphorus loads, the dominant nonpoint source in the Basin. Cost-effective phosphorus reduction strategies need reliable treatment techniques beyond basic cropland and waste management practices. The Lake Champlain Basin Agricultural Watersheds National Monitoring Program (NMP) Project evaluates the effectiveness of livestock exclusion, streambank protection, and riparian restoration practices in reducing concentrations and loads of nutrients, sediment, and bacteria in surface waters. Treatment and control watersheds in northwestern Vermont have been monitored since 1994 according to a paired-watershed design. Monitoring consists of continuous stream discharge recording, flow-proportional sampling for total P, total Kjeldahl N, and total suspended solids, grab sampling for indicator bacterial, and land use/agricultural monitoring. Strong statistical calibration between the control and treatment watersheds has been achieved. Installation of riparian fencing, protected stream crossings, and streambank bioengineering was completed in 1997. Early post-treatment data suggest significant reduction in P concentrations and loads and in bacteria counts in the treated watershed. Monitoring is scheduled to continue through 2000. PMID- 11379131 TI - Water quality characteristics of forest rivers around Lake Biwa. AB - Forest river surveys were carried out at upper streams of several rivers in the Lake Biwa watershed to understand the water quality characteristics of the rivers, and to find out their relationships with forest features such as geographical, geological and vegetational data. The results showed: (1) Forests have some purification functions for nitrogen and organic matter, but become sources for most of ionic species. (2) Main mineral species in forest rivers are Ca2+, Mg2+ and Na+, HCO3-, CO3(2-), Cl-, SO4(2-) and SiO2. (3) Loading from forests was 0.4-7 kg/km2/d for TN and 0.01-0.3 kg/km2/d for TP. (4) River quality reflects the properties of each forest, and is unique to the place, especially in ionic species such as Ca2+ and Cl-. (5) A cluster analysis successfully categorized ionic components into several groups. PMID- 11379132 TI - Impacts of agrochemical fertilizer on the aquatic environment of paddy fields in Vietnam. AB - To evaluate the impacts of agrochemical fertilizer application on the aquatic environment of paddy fields in tropical regions, 7.04 ha of paddy field situated in northern Vietnam were selected as a study area. The fate of nutrient constituents was surveyed through a questionnaire as well as analytical observation. Taking the major environmental components of the paddy field into account, a mass-balance flow regarding fertilizing constituents such as nitrogen and phosphate was built up and used to estimate the contribution of fertilizer to paddy field water pollution. In the mass-flow analysis, the randomness of fertilizer used by local farmers is incorporated within the conventional input output model. For the control volume of soil in 0-40 cm below paddy plot surface, the estimated average concentrations in soil were 1.8 ppm for inorganic nitrogen and 1.3 ppm for inorganic phosphate on the assumption of 50% non-excessive probability. From the reasonable consistency of the deduced concentrations of nutrients with observed ones, not only the mass balance of fertilizing substances but also the impacts of fertilizer on the paddy field aquatic environment in Vietnam could be understood. On the results from nutrient mass-balance analysis, it was found that 11.3-13.3 kg N/ha would percolate into the underground aquifer that is linked to surface channel flow in the vicinity of the paddy plots and affect the irrigation water quality. The nutrient portion in the materials harvested as grain and straw occupied 58% of total dosed nitrogen and 75.6% of total supplied phosphorous. PMID- 11379133 TI - Relating oxygen demand to flow: development of an in situ sediment oxygen demand measurement device. AB - This paper describes the continuation of an investigation into the effects of increased flow on the dissolved oxygen concentration in the River Maun, UK. A previous investigation indicated that an oxygen demand additional to that in the water phase is observed during and directly after intermittent discharges. Simplified modelling of the river indicated that the additional oxygen demand was possibly linked to the disturbance of sediment during such events. A device has been developed which can be used in situ to measure the effect of flow velocity (shear stress) on the additional demand. Preliminary tests have shown that the inferred demand is significant. It is proposed that the in situ device now be used to measure the oxygen demand of the bed material at a number of locations and flow speeds and that a relationship between the additional demand and velocity be established, with the aim of incorporating such a relationship into a simplified model. PMID- 11379134 TI - Physico-chemical and biological characteristics of thermal springs in Koycegiz and Dalaman basins in south-western Turkey and recommendations for their protection. AB - Many hydrogeological researches exist on the thermal springs of Turkey but limnological researches are very deficient. The results of the limnological research of the thermal springs in the basin of the meromictic lake Koycegiz and Dalaman Basin are as follows; the thermal springs are of euthermal or chliarothermal types. In the thermal springs thermotolerant diatoms constituted the major plankton flora. Recommendations for the protection of thermal springs are given according to the results of this research. PMID- 11379135 TI - Impact of atmospheric deposition on the headworks of a wastewater treatment plant -a case study. AB - Specialized sampling equipment and ultra-clean analytical methodology were employed to quantify the concentrations or fluxes of mercury (Hg), cadmium (Cd) and polychlorinated biphenyl (PCBs) in ambient air, precipitation, runoff, sanitary sewer, and wastewater treatment plant (WWTP) influent. The relationship between the atmospheric deposition and runoff on controlled surfaces were explored for the three pollutants. The impact of the atmospheric deposition and runoff to the headwork loading of the WWTP were investigated. Atmospheric deposition was found to be the primary source of the mass of Cd, Hg, and PCBs in runoff from the controlled surfaces. Neither atmospheric deposition nor the runoff was the main sources of the three pollutants to the Detroit Wastewater Treatment Plant (WWTP). Wet weather flow contributes the main portion of the Cd, Hg, and PCBs loading to the WWTP. PMID- 11379136 TI - Simulation of hyporheic self-purification in rivers: the assimilative capacity of proteins. AB - For the biochemical study of assimilation capacity (AC), two hypotheses were put forward (three levels of response, the hyporheon being the main site of self purification). A physical model was designed, simulating the hyporheon and its response to increasing inputs of proteins was studied. The analyses evidence a low accumulation of proteins in the hyporheic biofilms for daily flows ranging from 0.1 to 20 mg/day. The percolation of proteins stimulates the proteolytic activity (increase in Vm) whereas the Km shows no significant change; the assimilation capacities thus stimulated are vastly superior to the daily loads introduced into the system and explain the low accumulation of proteins. This stimulation concerns only a shallow active zone < 2 cm. For the future, a method of localising zones of hyporheic infiltrations (micro-piezometry, tracing, etc.) should be developed to monitor in situ the action of inputs of organic matter (OM) to estimate the AC of the river. PMID- 11379137 TI - A GIS planning model for urban oil spill management. AB - Oil spills in industrialized cities pose a significant threat to their urban water environment. The largest city in Canada, the city of Toronto, has an average 300-500 oil spills per year with an average total volume of about 160,000 L/year. About 45% of the spills was eventually cleaned up. Given the enormous amount of remaining oil entering into the fragile urban ecosystem, it is important to develop an effective pollution prevention and control plan for the city. A Geographic Information System (GIS) planning model has been developed to characterize oil spills and determine preventive and control measures available in the city. A database of oil spill records from 1988 to 1997 was compiled and geo-referenced. Attributes to each record such as spill volume, oil type, location, road type, sector, source, cleanup percentage, and environmental impacts were created. GIS layers of woodlots, wetlands, watercourses, Environmental Sensitive Areas, and Areas of Natural and Scientific Interest were obtained from the local Conservation Authority. By overlaying the spill characteristics with the GIS layers, evaluation of preventive and control solutions close to these environmental features was conducted. It was found that employee training and preventive maintenance should be improved as the principal cause of spills was attributed to human errors and equipment failure. Additionally, the cost of using oil separators at strategic spill locations was found to be $1.4 million. The GIS model provides an efficient planning tool for urban oil spill management. Additionally, the graphical capability of GIS allows users to integrate environmental features and spill characteristics in the management analysis. PMID- 11379138 TI - The nonionic surfactant pollution profile of Israel Mediterranean Sea coastal water. AB - Anionic and nonionic surfactants, as core components of detergent formulations, contribute significantly to the pollution profile of sewage and wastewaters of all kinds. In Israel about 15% of the total amount of ca. 4 x 10(8) m3/year of sewage is discharged, directly, or via receiving streams/rivers, into the Mediterranean Sea. Based on our previous findings that about 85% of the nonionic surfactants in the country's sewage are nonbiodegradable alkylphenol-based ethoxylates, we have undertaken this study, aiming at mapping the receiving eastern Mediterranean seawater with respect to its nonionic surfactant pollution profile. The total concentrations of nonionic surfactants were found--via reverse phase HPLC determinations--to be within the range of 4.2-25.0 ppb in seawater samples taken 2-3 m off the coastline at those locations where sewage-containing streams flow into the sea. Thus, neither the existing sewage treatment facilities nor natural biodegradation processes in receiving surface water systems are capable of avoiding this coastal water pollution. The potential estrogenic health risk of such concentrations of the anthropogenic EPEOs is dependent, among other things, on their specific homological distribution, biodegradation rate (slower for those having > 10 EO units) and survival. PMID- 11379139 TI - Soil characteristics and agrichemicals in groundwater of the midwestern United States. AB - A comprehensive set of soil characteristics were examined to determine the effect of soil on the transport of agrichemicals to groundwater. This paper examines the relation of soil characteristics to concentrations and occurrence nitrate, atrazine, and atrazine residue from 99 wells completed in unconsolidated aquifers across the Midwestern United States. Soil characteristics that determine the rate of water movement were directly related to the occurrence and concentrations of nitrate and atrazine in groundwater. The substantial differences in the relations found among soil characteristics and nitrate and atrazine in groundwater suggest that different processes affect the transformation, adsorption, and transport of these contaminants. A multi-variable analysis determined that the soil characteristics examined explained the amount of variability in concentrations for nitrate (19%), atrazine (33%), and atrazine residue (29%). These results document that, although soils do affect the transport of agrichemicals to groundwater, other factors such as hydrology, land use, and climate must also be considered to understand the occurrence of agrichemicals in groundwater. PMID- 11379140 TI - Input pathways and river load of pesticides in Germany--a national scale modeling assessment. AB - A model has been developed which estimates the magnitude and the spatial distribution of pesticide losses from diffuse sources (surface run-off, tile drains and spraydrift) into surface waters for the entire area of Germany. The cumulative annual losses of 42 active ingredients applied to 11 field crops, orchards and vineyards are calculated for river basins in Germany based on grid maps with a resolution of 1 x 1 km2. The model validation showed a sufficient degree of accuracy of the model results compared to measured pesticide loads in 13 small catchments. According to the model results the pesticide input from diffuse sources into surface waters amounted to 13.8 t in 1994 aggregated for the entire area of Germany. Input via surface runoff contributed 9.1 t while tile drainage was 1.4 t and spraydrift 3.4 t respectively. Alongside the model calculations empirical data of the pesticide load of rivers in Germany are presented. A comparison of the measured river loads with the modeled inputs from non-point sources leads to the conclusion that in most regions of Germany the largest portion of the load is due to the input from farm effluents. PMID- 11379141 TI - Characteristics of primary production in a eutrophicated bay. AB - The primary production of phytoplanktons provides organic matter in high concentration in eutrophicated Hakata bay in Japan, even during the winter season in spite of low water temperature. Phytoplanktons may have the biological capabilities to maintain activities of photosynthesis under unfavorable conditions, and these capabilities affect water quality in the bays. In this study, characteristics of primary production were analyzed with a simple box-type ecosystem model. We introduced a concept of efficiency for absorption of sunlight energy to our simulation model of water quality to explain rather high growth rates of phytoplanktons in low sunlight intensity. Through simulation with a box model, we found that the efficiency of primary production in winter is higher than that in summer. It was suggested that the organic pollution comes from dissolved organic carbon (DOC) throughout the year and that the DOC is originated from the primary production of phytoplanktons in biological response of the seasonal change of ambient conditions. PMID- 11379142 TI - Deriving robust operating rules for the interim control of the Kirazdere reservoir. AB - A methodology is developed for deriving robust operating rules for the interim control of a multipurpose reservoir. The methodology includes the generation of policies using optimisation models, the derivation of robust operating rules using artificial neural networks and the evaluation of the identified policies through simulation. The approach developed is demonstrated through an application to the Kirazdere Reservoir located on the Kiraz River in Turkey. PMID- 11379143 TI - A water quality index for contact recreation in New Zealand. AB - We surveyed the opinions of 16 water quality experts in order to develop a water quality index for contact recreation in freshwaters in New Zealand. The index was developed by postal surveys using the Delphi method, involving feedback of information to the panel members at each iteration. Determinands selected for use in the index were as follows: faecal bacterial indicators (faecal coliforms or E. coli), pH, Munsell colour, visual clarity indicators (black disc visibility or turbidity), and nutrients promoting nuisance growths (filtered BOD5, and dissolved forms of phosphorus and nitrogen). "Sub-index" curves relating suitability-for-use to these water quality determinands have been developed. The mean ("consensus") sub-index curves can be used to interpret water quality data in terms of suitability-for-use scores. We advocate using the lowest suitability for-use score for a water as its overall index value for contact recreation. Thus the water body's suitability-for-use is determined by its "poorest" characteristic. The index is now ready to be tested by water managers for its utility in state-of-environment reporting. PMID- 11379144 TI - Normalising impacts in an environmental systems analysis of wastewater systems. AB - In an environmental systems analysis of four wasterwater systems, the environmental aspects were prioritised by normalisation of predicted impacts from the studied systems to the total impacts from society. Priority Group 1 (highest priority) consisted of discharges (flows) of nitrogen, cadmium, lead and mercury to water, recycling of nitrogen and phosphorus to arable land and flows of heavy metals to arable land. A conventional wastewater system (A) was compared to irrigation of energy forest with biologically treated wastewater (B), liquid composting of toilet wastewater (C) and a conventional system supplemented with urine separation (D). Analysing the aspects in priority group one, systems B-D improved the management of plant nutrients and decreased the flow of heavy metals to water, while the flow to arable land increased, especially for system B. The suggested method is useful in municipal environmental planning and when choosing a wastewater system. PMID- 11379145 TI - Evaluation of appropriate system for reclaimed wastewater reuse in each area of Tokyo using GIS-based water balance model. AB - The appropriate type of reclaimed wastewater reuse system in each area of Tokyo was evaluated from the aspect of economic efficiency, using a GIS-based water balances model. The following four reclaimed wastewater reuse systems and conventional waterworks and sewerage system were evaluated; "Rain water storage and use system", "Onsite wastewater treatment and reuse system", "Sewage treatment and reuse at an intermediate point on the sewer pipe" and "Treated water supply system in sewage treatment plant". In the case that we install them to office and residential buildings, the supplied volume by reclaimed wastewater reuse systems is 693 thousands m3/d, this corresponds to 15% of total water demand in the area. Furthermore, the effects of the following scenarios brought about by technological innovation in water treatment were investigated; the case that flush water in toilet and wastewater from kitchen are also available as source in a "onsite wastewater treatment and reuse system" and the case that reclaimed water is used for laundering in residential buildings. When reclaimed water is used for laundering in residential buildings, the supplied volume by these systems increases to 814 thousand m3/d in the case that these systems are installed to office and residential buildings. PMID- 11379146 TI - Waste design and source control lead to flexibility in wastewater management. AB - The concepts of waste design and source control are introduced as contracts between wastewater handling agencies and consumers and consumers and industry respectively. Waste design serves to produce waste streams that are optimized in composition and time sequence for easy transport and treatment. Source control makes services and installations available, which allow the consumer to adhere to the specification of waste design. Increased delegation of responsibility to the consumer is expected to result in more flexible wastewater handling systems. PMID- 11379147 TI - River Water Quality Model no. 1 (RWQM1): III. Biochemical submodel selection. AB - The new River Water Quality Model no. 1 introduced in the two accompanying papers by Shanahan et al. and Reichert et al. is comprehensive. Shanahan et al. introduced a six-step decision procedure to select the necessary model features for a certain application. This paper specifically addresses one of these steps, i.e. the selection of submodels of the comprehensive biochemical conversion model introduced in Reichert et al. Specific conditions for inclusion of one or the other conversion process or model component are introduced, as are some general rules that can support the selection. Examples of simplified models are presented. PMID- 11379148 TI - Interaction analysis of stakeholders in water supply systems. AB - An analysis of the characteristic goals, strategies and rules of behavior of relevant stakeholders allows the efficacy and potential risks of past and current engineering and management concepts to be estimated. The study is driven by the observable shift from security to cost-centered strategies by water utilities and the difficulties of balancing technical and financial needs in an uncertain future. Its benefits include a methodology with a twofold result. With the aid of domain knowledge from experts involved in a participatory process, the interactions of a subset of stakeholders are quantified and documented in a rule catalog. This leads to an improved understanding of their decision-making rules. An agent-based model comprising these stakeholders' rules of behavior in subsequently development. Once the model is validated with data sets from a real utility, multiple-scenario testing helps to explore different strategies and can be used to generate ideas for developing flexible management and design schemes. Despite the complexity of the system described, simple model rules which are repeated annually can replicate the general development of both capacity and cost related parameters. Scenario simulations show the effects of different management strategies on key parameters such as capacity, water price and financial debt. PMID- 11379149 TI - Teaching future professors how to teach. AB - This paper describes a course designed to provide hands-on teaching experience to future professors and to incorporate techniques for more effective teaching. A team of Ph.D. candidates, under the direction of a senior faculty member, prepared a new course from beginning to end and then offered it to a class of graduate students. The course was developed using the unit map concept so that the presentations by the five student-instructors complemented and built upon one another. Immediately after each class, feedback was given to the student instructors by the faculty advisor and the other student-instructors. Review of video tapes of the lecture reinforced this feedback. At the completion of the course, both students and student-instructors were surveyed as to the effectiveness of the course and the student-instructors. This teaching experience and the feedback obtained from the surveys will be invaluable to the student instructors in their future development. PMID- 11379150 TI - Substrate interactions during aerobic biodegradation of methane, ethene, vinyl chloride and 1,2-dichloroethenes. AB - Intrinsic biodegradation of trichloroethene and 1,1,1-trichloroethane in groundwater at a Superfund site in California has been observed. An anaerobic zone exists in the area closest to the source location, yielding the expected complement of reductive dechlorination daughter products, including cis-1,2 dichloroethene (cis-DCE) and vinyl chloride (VC). Significant levels of methane and ethene were also generated in the anaerobic zone. The groundwater returns to aerobic conditions downgradient of the source, with methane, ethene, VC, and several other compounds still present. Attenuation of VC in the aerobic zone suggests that it is being biodegraded. In this study microcosms were used to evaluate the role of methane and ethene as primary substrates for aerobic biodegradation of VC. Biodegradation of VC was fastest in the bottles containing ethene, with 40 mumol of VC consumed over a 150 day period, compared to approximately 15-20 mumol with methane or a mixture of methane and ethene. VC did not noticeably inhibit ethene biodegradation but did slow the rate of methane use. Methane inhibited ethene metabolism, which apparently caused a reduction in VC biodegradation when methane was present with ethene. These results suggest that ethene plays an important role during in situ natural attenuation of VC under aerobic conditions. Microcosms were also set up with VC alone. Following a 75 day lag period. VC consumption began and subsequent additions were consumed without a lag, suggesting the presence of organisms capable of using VC as a growth substrate. After providing VC alone for nearly 400 days, aliquots of the enrichment culture were used to evaluate its ability to biodegrade cis- and trans DCE. Both compounds were readily consumed, although addition of VC as the primary substrate was needed to sustain biodegradation of repeated additions. This result suggests that organisms capable of using VC as a sole substrate may play an active role in aerobic natural attenuation of DCEs. PMID- 11379151 TI - Effects of subsurface heterogeneity on natural bioremediation at a gasoline spill site. AB - A test cell of 3-m by 6-m located at the mid-point of a gasoline spill site was selected to test the hypothesis that the rate of hydrocarbon biodegradation is influenced by the spatial distribution of the electron acceptors, aqueous geochemistry, and microbial population. Multilevel samplers (MLSs) were installed at four corners of the test cell for groundwater sampling. Sampling ports were placed at 0.3-m intervals from 1.5 to 4.8 m below land surface (bls). A 0.91-m by 12.7-cm sediment core (from 3.3 to 4.2 m bls) in the center of the MLSs was collected. The core was cut into 7 sections, and each was used for sediment extractions, microbial enumeration, grain size distribution, and microcosm studies. Groundwater analytical results indicate that iron reduction was the dominant biodegradation process within this test cell. Iron-reducing process caused the preferential removal of certain compounds. Microbial enumeration results show that the distribution of microbial population varied with depth and sediment materials. Lower microbial population was observed in those sections with higher portion of clayey materials. The less permeable materials would limit the bacterial transport, decrease the bioavailability of Fe(III) to iron-reducing bacteria, and thus cause the low biodegradation activity. Results suggest that using blended sediments for biodegradation rate measurements may provide misleading results. PMID- 11379152 TI - Oxidation of polycyclic aromatic hydrocarbons by ozone in the presence of sand. AB - A series of soil slurry experiments was performed to investigate the characteristics of PAHs removal by ozone in various conditions. Gaseous ozone was bottled into the aqueous phase in the presence of soil contaminated by PAHs. The effects of soil media, OH radical scavengers, ozone dosage, and humic acid were examined at the given experimental conditions. There exists a substantial difference in the removal of PAH according to the types of soil media tested. Baked sand showed the highest removal efficiency compared to the others. The descending order of removal rate was: BS > S > GB. This is considered to be due to the OH radical effect produced by catalytic reactions of ozone with the reactive site on the and. This is qualitatively proved by the experiment of scavenging OH radicals using tert-butanol. The comparison of half-lives of ozone in sand and glass bead columns further supports this hypothesis. It was found that about 22% of enhancement of phenanthrene destruction was accomplished by OH radicals produced by the catalytic ozone decomposition. The rate of ozone consumption for the phenanthrene oxidation was obtained as 1.88 mg/mgO3/min. PMID- 11379153 TI - Novel boom/skirt systems for improvement of water quality in estuarial impoundments subject to saline influx. AB - The recent popularity of esturial barrage construction in the UK has been driven largely by commercial and amenity interests arising from the need to rehabilitate tracts of derelict urban land. Formation of a permanent water body to replace the tidal regime hides unsightly mud banks and facilities ready access for water based recreation. Unfortunately, in most cases, and presumably arising from upstream extreme flood risk assessment, the barrier is designed to overtop under the highest (spring) tidal peaks so allowing a substantial influx of heavier saltwater into the impoundment on a regular basis. This might well substantially exceed influx via any navigational lock operations. The problem created by the salt water is that its greater density causes stratification in the impounded water body. In times of low freshwater flow little mixing takes place and the normal action of the wind, in circulating and aerating the water body, will only extend within the lighter and often thin surface freshwater layer. The consequence is that little reoxygenation of the lower layers takes place leading to depressed dissolved oxygen levels and possible anoxia. Sluice provision in the barrage may be ineffective in expelling the saline layers, so this water remains near stagnant and deteriorates in quality due to sediment oxygen demands until a major fluvial flood, or new saline influx, is able to effectively mix the waters. Conventional continuous surface releases through fish pass and/or by crest overspill ensure the preferential release of the freshwater rather than the problematic saltwater. Retrofitting of novel low cost floating boom/skirt baffle system is proposed here for alleviation of the problem. PMID- 11379154 TI - Complete remediation of PCE contaminated unsaturated soils by sequential anaerobic-aerobic bioventing. AB - Bioventing principles have been applied to completely dechlorinate tetrachloroethylene vapors in the unsaturated zone in a sequential anaerobic aerobic pattern. The aerobic step yields trans-DCE and VC as PCE reductive dechlorination byproducts, while TCE and cis-DCE are observed as intermediates. The aerobic step results in rapid oxidation of the VC and trans-DCE to carbon dioxide. Hydrogen was delivered in the gas phase as a reducing agent for the anaerobic step at levels of 1%, and oxygen at 4.2% was used as an electron acceptor in the aerobic step. PCE and VC half lives in the anaerobic and aerobic steps respectively, where less than 10 min. PMID- 11379155 TI - River Water Quality Model no. 1 (RWQM1): case study. I. Compartmentalisation approach applied to oxygen balances in the River Lahn (Germany). AB - A case study on the application of the River Water Quality Model No. 1 (RWQM1) is presented in order to illustrate the importance of modelling a sediment compartment for an ecologically meaningful assessment of the impact of wastewater effluents and combined sewer overflows. The focus of this case study is on the compartmentalisation approach of the RWQM1 that makes such a description possible. In contrast to this, a strongly simplified biochemical submodel is used that considers only oxygen and dissolved substrate. The object of the case study is the River Lahn, a moderately polluted 5th order stream in Germany, for which the connectivity of surface/subsurface flows and mass fluxes within river sediments have been intensively investigated. The hyporheic flow between a downwelling and upwelling zone of a riffle-pool sequence has been studied with the aid of tracer experiments and continuous records of water constituents. High diurnal fluctuations of oxygen travelled to considerable depth of the sediment and oxygen in the interstitial water decreased considerably while travelling through the riffle. Starting with the implementation of a strongly simplified version of the biochemical part of the RWQM1, but with the consideration of a sediment pore water compartment in addition to the water column compartment, a calibration procedure is performed using tracer data from the water column and the sediment. The calibrated model is then used to study the system response to wastewater treatment plant effluent and combined sewer overflow emissions. The modelling approach makes it possible to quantify the sediment oxygen demand and the spatial and temporal extent of sediment zones with oxygen depletion. However, the spatially averaged approach does not account for inhomogeneities in the sediment. It is shown that for this river with its alluvial coarse sediments even moderate emissions from sewerage systems may be high enough to drop sediment oxygen concentrations to low levels while those in the surface flow remain close to saturation. Similarly, it is demonstrated that combined sewer overflows may cause anoxic sediment oxygen conditions for extended time periods. The implications for ecologically sound river water quality modelling and for specific quality objectives are discussed. PMID- 11379156 TI - River Water Quality Model no. 1 (RWQM1): case study II. Oxygen and nitrogen conversion processes in the River Glatt (Switzerland). AB - Various simplifications of the river water quality model no. 1 are applied to data sets from the river Glatt in Switzerland. In a first application, the biomass responsible for nitrogen and oxygen conversion processes is quantified based on known reaeration rates, measured concentrations of ammonia, nitrite and oxygen and assumed growth parameters of algae and bacteria. In a second application, the model is extended to calculate chemical equilibria of inorganic carbon compounds dissolved in the water and daily variations in pH. The influence of partially unknown inflow concentrations and of calcite precipitation on fluctuations in electrical conductivity and pH are discussed. In the last model, the processes of growth of sessile algae and bacteria, detachment of algae, and grazing by benthic organisms are introduced. Due to lack of data for quantifying these processes, this last model application is speculative. Nevertheless, it is interesting because it shows a direction to which river water quality modelling would have to proceed in order to increase its predictive capabilities. PMID- 11379157 TI - Urban drainage redefined: from stormwater removal to integrated management. AB - Even though urban drainage has been practised for more than 5000 years, many challenges arising from growing demands on drainage still remain with respect to runoff quantity and quality; landscape aesthetics, ecology and beneficial uses; and operation of existing urban wastewater systems. Further advances can be achieved by adopting an integrated approach, optimal operation of the existing infrastructure, advanced pollution and runoff source controls, improved resilience of receiving waters, and adaptive water management. The specific research needs include new technologies and strategies for stormwater management, advanced treatment of urban wet-weather effluents, and tools for analysis and operation of drainage systems. High diversity of demands on, and region/site specific conditions of, urban drainage shapes the role of urban drainage experts- as mediators among the many stakeholders and fields involved. PMID- 11379158 TI - Risk based urban watershed management under conflicting objectives. AB - Ecological impairment and flooding caused by urbanization can be expressed numerically by calculating the risks throughout the watershed (floodplain) and along the main stems of the streams. The risks can be evaluated in terms of the present and/or future. This article describes the methodologies for ascertaining the risks in the Geographical Information Systems (GIS) environment. The objectives of urban flood controls and ecological preservation/restoration of urban waters are often conflicting and, in the past, the sole emphasis on flood control led to destruction of habitat and deterioration of water quality. An optimal solution to these two problems may be achieved by linking the risks to the concepts of risk communication, risk perception, and public willingness to pay for projects leading to ecological restoration and ecologically sustainable flood control. This method is appropriate because, in each case, public funds are used and the projects require approval and backing of policy makers and stakeholders. This article briefly describes a research project that attempts to resolve the conflict between the flood protection and stream ecological preservation and restoration and suggests alternative ways of expressing benefits of urban stream flood control and restoration projects. PMID- 11379159 TI - Radar based rainfall forecast for sewage systems control. AB - There has been an increasing demand for accurate rainfall forecast in urban areas from the water industry. Current forecasting systems provided mainly by meteorological offices are based on large-scale prediction and are not well suited for this application. In order to devise a system especially designed for the dynamic management of a sewerage system the "RADAR" project was launched. The idea of this project was to provide a short-term small-scale prediction of rain based on radar images. The prediction methodology combines two methods. An extrapolation method based on a sophisticated cross correlation of images is optimised by a neural network technique. Three different application sites in Europe have been used to validate the system. PMID- 11379160 TI - Simple mass balance approach for assessment of flood control sumps in an urban watershed: case study of heavy metal loading. AB - Levee sump systems are used by many riverine communities for temporary storage of urban wet weather flows. The complex hydraulics and transport of stormwater pollutants in sump systems, however, have not been systematically studied. The objective of this work is to present a case study, utilizing a relatively simple and low-cost methodology, for assessing the hydraulic performance of flood control sumps in an urban watershed. Two sumps of highly variable physical and hydraulic characteristics were selected for analysis. HEC-1 software was used to estimate the flow hydrograph for each outfall to a sump as part of the overall flow balance, resulting in a total runoff hydrograph for a precipitation event. To validate HEC-1 results, a water balance was used to estimate the total runoff using sump operational data. The results suggest that HEC-1 calculation provide a satisfactory estimate of the total runoff and its time-distribution to the sump. The hydraulic model was then used to estimate nonpoint loads of selected heavy metals to the sump and to the river. Although flow of stormwater through a sump system is regulated solely by flood-control requirements, these sumps may function as sedimentation basins that provide purification of stormwater. An example calculation of removal of heavy metals in a sump using a mass balance approach is presented. PMID- 11379161 TI - The erosion of organic solids in combined sewers. AB - Many studies undertaken on urban catchments show, thanks to indirect approaches, that the contribution of eroded sewer sediments to pollution of combined sewer overflows is significant and highly organic. An in situ study of the erosion of sewer sediments has been implemented to validate those results with a direct approach and to observe the processes of erosion. Two experiments have been carried out on a 150 m length of combined sewer in "Le Marais" catchment in Paris, in order to determine the rate of erosion and the nature of the particles eroded by an injection of drinking water in the sewer system. Hydraulic and quality parameters have been measured in situ. Those injections have shown that the rate of erosion is important (maximum rate of 146 g/s) at each stage of the injection, which has been conducted in three stages with a maximum flow of 370 m3/h. The erosion does not only occur locally but happens along the entire length of the section even at low shear stresses (0.5 N/m2). The eroded particles are highly organic (VS = 54-86%) and their loads in volatile solids, COD, BOD5 decrease as the flow increases. So, this work confirms, by direct measurements, that eroded sewer sediments are a significant source of organic matter that contribute to combined sewer overflow. PMID- 11379162 TI - [New series: Pain and pain relief a neglected specialty]. PMID- 11379163 TI - [Wrong focusing in the debate on euthanasia. Consider patient's situation and motifs, not the predetermined view for and against among healthy debaters]. PMID- 11379164 TI - [Pain--life's dreaded companion. Cultural history of algology and the most important achievements]. AB - Pain is a comprehensive phenomenon which concerns not only physiology, biochemistry and molecular biology but also music, art, religion, philosophy and everyday life. The early contributions to the development of the specificity theory is described, as are some of the important discoveries in the history of algology, such as Baillou's description of rheumatism, Heberden's description of angina pectoris and the development of the gate control theory. Despite a number of important discoveries and a much improved knowledge in basic science pain is still the dreaded companion of mankind. PMID- 11379165 TI - [Postoperative pain an unnecessary suffering. A model of "emergency pain relief" implemented in Orebro]. AB - Despite unprecedented interest in the understanding of pain mechanisms and pain management, a significant number of patients continue to experience unacceptable pain after surgery. An acute pain service (APS) has to include regular pain assessment and documentation, "make pain visible", and bedside teaching of nurses in order to provide safe and cost-effective analgesia. The acute pain team in Orebro includes anaesthesiologists, surgeons, specialist acute pain nurses as well as ward nurses. A combination of less invasive surgery, effective analgesia and strategies for rehabilitation can reduce post-operative morbidity and shorten hospitalization. PMID- 11379166 TI - [Pain and pain relief during the neonatal period. Early pain experiences can result in negative late-effects]. AB - The neonate has a functional nociceptive system. However, recent research suggests that infants may be more vulnerable to the negative effects of pain than older children and adults. Apart from short-term effects, untreated pain may also have long-term effects, which may later affect neurological development, including the reaction to pain. Despite convincing evidence from recent research, the neonate is still subjected to painful procedures, even surgery, without adequate treatment. PMID- 11379167 TI - [Amino acid administration counteracts hypothermia during anesthesia]. AB - General anesthesia induces hypothermia due to decreased metabolic rate and impaired thermoregulation. Adverse effects of hypothermia are common. Many warming devices are in use to prevent heat loss, but little attention has been paid to stimulating the body's own heat generation. NIT, nutrient-induced thermogenesis, signifies the concept that all nutrients raise energy expenditure. The greatest thermic effect is ascribed to amino acids and proteins, 30-40%. During anesthesia the thermogenic effect of i.v. amino acids was increased. The results support the existence of an inhibitory action normally exerted by central thermosensors in order to prevent hyperthermia. During anesthesia, central thermosensors are impaired, and hence amino acid thermogenesis is exaggerated, which may prevent hypothermia. PMID- 11379168 TI - [Cooling as treatment of birth asphyxia]. AB - In about 10 per cent of children with cerebral palsy, the most probable cause is birth asphyxia. The brain injury following birth asphyxia evolves in part over hours or days--as a secondary process. This opens a 'window of opportunity' for intervention. Decreasing the temperature by as little as 3 degrees C appears to be effective. It is possible to identify babies with a 50 per cent risk of death or survival with cerebral palsy within 3 to 6 hours after birth. Pilot trials of cooling suggest that the side-effects of cooling can be managed. Randomized controlled trials are underway. Obtaining informed consent from the parents for these trials represents a particular challenge. PMID- 11379169 TI - [Increased consumption of drugs among the elderly results in greater risk of problems]. AB - With aging comes an increasing prevalence of diseases and symptoms that frequently require pharmaceutical treatment. However, aging also brings about bodily changes that result in increased effects and prolonged action of many drugs. Multiple drug use--often termed polypharmacy--seen in many elderly individuals, is the most important risk factor for adverse drug reactions (ADR) and increases the risk of drug interactions and poor compliance. ADR's are responsible for about 10% of all hospital admissions of elderly patients. The drugs most commonly involved are cardiovascular, psychotropics and anti inflammatory agents. Many of these ADR's are dose-dependent and preventable. Drug use has increased over the last few years, largely thanks to the availability of new and effective agents. This calls for increased vigilance and prudence in prescribing for the elderly. PMID- 11379170 TI - [Incidence of surgery for hydrocephalus in adults surveyed: same number afflicted by hydrocephalus as by multiple sclerosis]. AB - The incidence of surgical treatment for adult hydrocephalus (older than 18 years) in Sweden from 1996 to 1998 was surveyed. The number of operations was 891 and the average incidence 3.36 operations per 100,000 inhabitants and year, varying regionally from 2.3 to 6.3. The mean age was 60 years (range 18-92), with no sex difference. Normal pressure hydrocephalus (47%) was most common, followed by communicating high pressure hydrocephalus (27%) and aqueductal stenosis (11%). 804 shunt operations (90%) and 67 ventriculostomies (7.5%) were performed, 2% were unclassified. PMID- 11379171 TI - [Regional anesthesia reduces perioperative morbidity and mortality]. PMID- 11379172 TI - [The greater the crowd, the greater the risk of accidents. How good is our disaster planning and who is to finance it?]. PMID- 11379173 TI - [To know own diagnosis or not--that is the question]. PMID- 11379174 TI - [Spinal diseases which disappeared. The Swedish Society of Spinal Diseases rejects the SBU report]. PMID- 11379175 TI - [Reflections on the SBU report. Acupuncture in the treatment of back and neck pain]. PMID- 11379176 TI - [A 84-year old woman "relatively" killed]. PMID- 11379177 TI - [Are there any hazards connected to cerebral computer tomography?]. PMID- 11379178 TI - [Obesity is a chronic disease--as asthma]. PMID- 11379179 TI - [Consider influenza vaccination in heart disease trials]. PMID- 11379180 TI - [Who puts the limits in primary health care?]. PMID- 11379181 TI - [Is allergy to electricity really a phobia?]. PMID- 11379182 TI - A presidential conversation. PMID- 11379183 TI - MedBytes. PMID- 11379184 TI - Reduce care or raise taxes. PMID- 11379185 TI - Labor pains. PMID- 11379186 TI - Asking the heart questions. PMID- 11379187 TI - No bundle of joy. PMID- 11379188 TI - When is enough enough? PMID- 11379189 TI - Stroke prevention essentials. PMID- 11379190 TI - Suicide in Travis County, Texas, from 1994 through 1998. AB - Suicide, the ninth leading cause of death in the United States, has not been adequately studied in Travis County, Texas. To describe the occurrence of completed suicide in Travis County, 426 cases of completed suicide from 1994 through 1998 were collected and the data were analyzed with the chi 2 test and the t test. The annual suicide rate in Travis County is 13.0 per 100,000. Caucasian males have the highest annual suicide rate among all races, followed by Hispanics and, then, blacks. The highest suicide rate was seen in persons aged 75 years and older. The second most frequent rate was for those aged 45 through 54 years. Guns are the most common means of suicide in both males and females (60.8% and 37.3%, respectively). A person who is Caucasian, male, and aged 75 years and older is at the highest risk. Reducing the availability of firearms may have a significant impact on the suicide rates in Travis County. PMID- 11379191 TI - [The Lijecnicki Vjesnik medical journal at the threshold of the third millenium]. PMID- 11379192 TI - [Bartonella henselae as the causative agent in cat-scratch disease: case report]. AB - In this article we reported typical clinical, primary skin lesion and regional lymphadenitis, and atypical, protracted fever and algic syndrome, characteristics of cat scratch disease (CSD) in a 21-year-old man (a student) from Zadar, Croatia. Laboratory parameters were in normal range. The histopathologic findings of affected lymph nodes included stellate caseating granulomas. By using IFA method a seroconversion of specific IgG antibodies (neg/1:512) and rise of IgM antibodies (1:160/ > 1:320) to B. henselae were detected in paired sera, and these serologic findings indicate on conclusion that B. henselae is probably etiologic agent of CSD in our patient. PMID- 11379193 TI - [Antimicrobial therapy of urinary tract infections]. AB - Urinary tract infections (UTIs), according to localization of infection, can be subdivided into urethritis, cystitis, prostatitis and pyelonephritis, according to type of infection into symptomatic, asymptomatic, acute (first or single), recurrent, chronic, complicated and uncomplicated. Clinical symptoms of cystitis and leukocyturia are sufficient reason for early initiation of a three-day empirical antimicrobial therapy of acute uncomplicated cystitis in young women. Urine culture should be performed prior to the initiation of antimicrobial therapy in pregnant women, diabetics, recurrent UTIs, in case of unsuccessful prior treatment and in patients with pyelonephritis. All symptomatic UTIs should be treated, as well as asymptomatic bacteriuria in pregnant women, diabetics, preschool children and prior to urologic-gynecologic surgery. In complicated UTIs it is especially important to determine and try to eliminate or at least put under control the factors that complicate UTIs. Antimicrobial therapy of UTIs includes fluoroquinolones, co-trimoxazole, betalactam antibiotics, aminoglycosides and nitrofurantoin, tetracyclines, macrolides, and azalydes in case of sexually transmitted diseases caused by Chlamydia trachomatis and Ureaplasma urealyticum. Cystitis is treated for 1, 3 or 7 days, asymptomatic bacteriuria 3-7 days, uncomplicated pyelonephritis 10-14 days, bacterial prostatitis 4-8 weeks, and chronic nonbacterial prostatitis 2-4 weeks. Recommended therapy for chronic and complicated UTIs is 7-14 days only in relapses and reinfections, and in some patients it can last for several weeks, up to 6 months. Chemoprophylaxis in recurrent uncomplicated UTIs should be employed for at least 6 months. PMID- 11379194 TI - [New information on the pathophysiology of atherosclerosis]. AB - The past decade has witnessed enormous progress in our understanding of the nature of this process. The development of an atherosclerotic plaque is a complex process which begins with endothelial dysfunction, the trigger for which are factors such as hypercholesterolemia, smoking, hypertension, hyperhomocysteinemia and impaired glucose metabolism. This dysfunction includes increased endothelial permeability to lipoproteins and other plasma constituents, which is mediated by NO, PDGF, prostacyclin, angiotensin II and endothelin; up-regulation of endothelial adhesion molecules including VCAM-1, ICAM-1, and selectins and migration of leukocytes and monocytes-macrophages in the subendothelial space mediated by oxidized LDL, MCP-1, PDGF and MCSF. The next step includes smooth muscle cells migration (stimulated by PDGF and TGF-beta), T-cell activation (mediated by TNF-alpha and IL-2), formation of foam-cells from macrophages (mediated by oxidized LDL, MCSF, TNF-alpha and IL-1) and platelet adherence and aggregation (stimulated by thromboxane A2, tissue factor etc). The smooth muscle cells form a fibrous cap which confers mechanical stability of the plaque and separates the lipid rich thrombogenic core from the lumen and circulating blood. Whether a plaque will remain intact and therefore stable or rupture and lead to thrombosis causing an acute coronary syndrome (MI, unstable angina pectoris) depends upon a number of factors, the most important of which is its composition. Plaque size plays only a minor role in determining risk of an acute coronary syndrome. Rupture of the fibrous cap occurs due to thinning of the cap caused by an influx and activation of macrophages which release metalloproteinases and other proteolytic enzymes (stimulated by inflammatory cells, particularly T lymphocytes). These enzymes cause degradation of the fibrous tissue of the cap which can result in thrombous formation and occlusion of the artery. Stable plaques have a thick fibrous cap, a small lipid core, and few inflammatory cells. In contrast, vulnerable plaques have a high lipid content, numerous inflammatory cells, and a thin fibrous cap with reduced collagen and vascular smooth muscle cells in it. Although vulnerable plaques are believed to account for only a small number of all coronary atheromas, they are responsible for most acute coronary events. PMID- 11379195 TI - [Sideropenic anemia in infants and toddlers]. AB - Iron-deficiency anemia is the most common anemia in infants. In the routine pediatric care this problem is encountered every day. Numerous factors in infancy (low birth weight, rapid growth, insufficient nutrition) are involved in the development of iron-deficiency anemia, and they must be considered when establishing diagnosis, counselling parents, and prescribing oral iron preparations. Data on 119 patients aged up to two years treated in the Division of Gastroenterology and Nutrition, Pediatric Department, University Hospital Centre Zagreb Salata between 1994 and 1999, were analyzed. We were prompted to do so because of great frequency of iron-deficiency anemia as one of associated diagnoses, and unfortunately often the only diagnosis requiring hospitalization. Out of 119 patients with iron-deficiency anemia, nine (7.7%) patients (four premature newborns and three from twin pregnancy) had to receive transfusion of erythrocyte concentrate due to very bad general condition and low red blood count, accompanied by clinical signs of anemic hypoxia. We also analyzed prenatal and perinatal history, socioeconomic living conditions of these children, i.e. their nutrition, and if they had previously received oral iron preparations. Some of the results, such as inadequate alimentation with flour, as well as insufficient prophylaxis of iron deficiency, which were found in most cases of severe anemia, point to the need of paying greater attention to this problem, better parents education, and more adequate screening for iron deficiency anemia. PMID- 11379196 TI - [Museum of the History of Hygiene in Croatia]. PMID- 11379197 TI - [Euthanasia]. AB - The paper gives a historical overview, and describes psychosociological constellation and tactics of the euthanasia movement. Further, medicosocial experiment and medicolegal engineering which preceded it and followed it are presented on the example of euthanasia in The Netherlands. Legal, ethical, formally logical and psychological arguments against euthanasia are presented. The article concludes with the opinion that it was the duty of the World Medical Association to protect medical profession with the procedure of demedicalization of euthanasia, and to appropriately punish the Royal Dutch Medical Society for breaking professional ethical codex in its essential part, which is the basis of the very identity of medical profession. PMID- 11379198 TI - [Hospitalization for systemic connective tissue diseases in Croatia in 1997: examples of the use of hospital morbidity registery data bases]. AB - By using the explanatory analysis methods, we examined a possible model for the utilization of the Croatian Hospital Morbidity Register data for decision making in the managerial process for national health system development. To build a draft informational model, data were selected on 843 cases hospitalized in 1997 for diseases M30-M36 (ICD-10) in 45 health institutions. This was used as an index of the diagnostically/therapeutically most demanding group of rheumatic autoimmune diseases. Treatment standards were established for our health system for the patients having an M30-M36 disease by classifying Croatia's health institutions into four clusters by intensity of hospitalization, and by analyzing demographic characteristics and the length of stay by disease entity. These standards could represent a good informational base for forming consensus group panels to deal with inpatient treatment problems of patients with systemic connective tissue diseases. PMID- 11379199 TI - [Psychological, emotional, spiritual and social aspects of pain]. AB - We are used to speak about the organic pain origin. Meanwhile in actual and old definitions of pain the other determinants, besides the organic one, are included. They are changing the pain threshold and the pain tolerance. Dame Cicely Saunders, who started the modern hospice movement of care for those in advanced stage of disease, describes the concept of "total pain" as involving organic, emotional, spiritual and social determinants. Such pain concept fits into the classical division of pain pathogenesis. The basic forms are: nociceptive, neuropathic and according to the old nomenclature psychogenic. The first one is defense reaction, the second one is due to pathology. The third one is very complex and this article is dealing with it. The patient's emotional reactions, especially the fear and anxiousness producing tension increase the pain. Therefore, different kinds of relaxation or attention distraction can help a lot in relieving the pain. Spiritual pain can be a very difficult symptom increasing the physical pain significantly. Everybody has spiritual needs and they are independent of religious orientation. The experience of life's and suffering's senslessness, personal valulessness and situational hopelessness lowers significantly pain tolerance. The selfconfidence, the confidence in other persons, in somebody over us has the opposite effect. And finally the pain is often easier to cope with in the presence of a friendly person. To be able to complain and to describe one's own fears, especially in connection with the significance of the actual pain, helps a lot. The support of the wider community also helps. We ought to approach the patient holistically, that means also observe the person, the human being suffering, hoping and wishing to be heard, and not to be left alone. PMID- 11379200 TI - [The Museum of the History of Hygiene and the Croatian Medical Society]. PMID- 11379201 TI - [Ortho-phthalaldehyde: a new disinfectant highly effective in disinfection of endoscopes]. PMID- 11379202 TI - [Bile duct injuries in laparoscopic cholecystectomy]. AB - Bile duct injuries are the most serious complications of laparoscopic cholecystectomy. Although the overall complications rate in laparoscopic cholecystectomy is significantly lower than in open cholecystectomy, bile duct injuries are more frequent. In this article we analyzed why and how they occur and if their prevention was possible. From May 1992 to May 1999, laparoscopic cholecystectomy was performed in 5651 patients by 17 surgeons and residents, with different experience but similar operative technique. In this period nine (0.16%) bile duct injuries occurred. Injuries were caused by seven operators. Number of operations by single surgeon before injury varied from 8 to 1477. Three patients with uncomplicated chronic cholecystitis had puncture or small bile duct laceration. Complete transection of main bile duct occurred in three patients and in one transection of right hepatic duct. The operative findings in two of them were acute cholecystitis, one had fibrosis in the triangle of Calot and one uncomplicated chronic cholecystitis. Bile duct strictures were found in two patients. Both had fibrosis in the triangle of Calot and in hepatic port. Dangerous pathologic findings were the main cause of injuries in 5 and inadequate surgical technique in 4 patients. In our opinion the education, critical state and experience are of main importance in avoidance and prevention of bile duct injures. We also plead for selective use of intraoperative cholangiography. In our opinion the intraoperative cholangiography has to be performed in all cases where the anatomy of the triangle of Calot is not clear. PMID- 11379203 TI - Industry and water security: overarching conclusions. AB - Fresh water is key to sustainable development. World Business Council for Sustainable Development members are addressing fresh water use "within the corporate fenceline". However, to address water issues "outside the corporate fenceline" will require creative new public-private partnerships. Government's role is to provide sound framework conditions that will encourage businesses to invest time, staff and resources to address vital fresh water issues. Industry is committed to best practice within its internal operations and is ready to enter into partnerships to address broader fresh water issues. PMID- 11379204 TI - Workshop 1 (synthesis): steps towards increased effectiveness in water use- possibilities and replicability. AB - Inevitable increases in human needs and wants leading to mounting water stress necessitate that each unit of water is used as efficiently as possible. Excessive use of water cannot be accepted. Similarly, the least possible impact on water quality is a must. Yet, regional development and urgent social and environmental objectives may motivate an allocation of scarce water resources where the output is lower as compared to what could have been accomplished with an alternative allocation. In essence, the effective use of water presumes a broad and long-term perspective on societal development and environmental sustainability. Presentations illuminated that technical and institutional arrangements must be applied to reach an effective use. In practice, although not ncessarily in policy statements, technical arrangements continue to have a pivotal position in this regard. PMID- 11379205 TI - The role of institutional arrangements for more efficient water resources use and allocation. AB - Water resources management requires an interdisciplinary approach to meet the challenges posed by ever increasing water demand and pollution. This paper discusses in which way appropriate institutional arrangements are complementary to technical interventions that aim to increase more efficient water use and allocation. Specific emphasis is given to water property rights as one of the determining factors in regard to the institutional framework. Issues such as the range of flexibility in designing property rights to water, the implications for pricing, enforcement and sanctioning mechanisms as well as for water trading and information needs are highlighted. PMID- 11379206 TI - Workshop 2 (synthesis): balancing water and social challenges through restructuring of industry. AB - The workshop discussed the possibility of breaking the link between economic growth and environmental damage, and options that would encourage industry and government to take steps to reduce emissions, with 8 catalysts for change identified. PMID- 11379207 TI - Towards sustainable development: catalysts for change in industrial water management. AB - "Business as usual is not an option." Water management practices are changing too slowly and the global "fresh water gap" continues to grow. Actions as must be taken by government and civic leaders. Industry also has a responsibility to act, but its inability to adequately meet the challenge to date points to the need for additional stimuli. The current water crisis can be mitigated by changes in behaviour and perceptions. This presentation will argue that specific catalysts are required to encourage and support a dramatic shift of behaviours and perceptions by industry leaders. The catalysts include: establish clear and objective rules; introduce water pricing; recognize "life cycle" costs; prioritize needs; reward small-scale solutions; nurture innovations; spread global best practices. With the help of the catalysts described herein, industry can take its place as a leader in effective water management in the 21st Century. In partnership with governments, international organizations and civic organizations, industry leaders can help meet the challenge of the water crisis while creating sustainable economic growth. PMID- 11379208 TI - Workshop 3 (synthesis): innovative processes in small scale agricultural production using water more effectively. AB - The workshop discussed the possibilities of innovative technologies to increase food production. Most of the concepts discussed are based on rainfall management, conservation and recycling. Simple technologies, adopted to local circumstances, can increase farm yields in small-scale agriculture. Small-scale rainfed agriculture is viable with intelligent utilisation of harvested rainfall. These small-scale technologies need however to be complemented with educational programs, incentives and trade to be broadly adopted and effective. The need for better documentation of results was also raised in the discussion. PMID- 11379209 TI - Preserving the (water) harvest: effective water use in agriculture. AB - The alternative to increasing the world's irrigated area by an estimated 30% to secure food security for all, seems to be limited irrigation expansion and consequently higher food prices and probably food shortages. This paper explores other options for ensuring food security. It discusses meaningful similarities between innovative approaches for land and water management in rainfed and irrigated agriculture. The focus is on innovative approaches to increase yields in sub-Saharan Africa and South Asia. Innovative technologies, such as improved tillage practices and water harvesting are important. But at least as important are the processes by which new agricultural practices are developed, improved and extended. In the end it comes down to human inventiveness. PMID- 11379210 TI - Workshop 4 (synthesis): responsibilities linked to "human rights approach" to water--rules and roles. AB - Thinking in terms of ethics and Human Rights raises issues of power and justice. It can be argued that there are indisputable human rights, e.g. when the construction of dams is considered, while 'commodification' of water and the 'market mentality' may suggest that everything can be traded. PMID- 11379211 TI - Water law and human rights--roles and responsibilities. AB - This paper focuses on the experience of South Africa in introducing water legislation based on human rights principles (in particular the National Water Act of 1998) and reflects on some practical implications for the implementation of water management in a country with limited water and financial resources. PMID- 11379212 TI - Workshop 5 (synthesis): feminization of water security. AB - Water management can be greatly improved if the capacities, skills and perspectives of women are promoted. PMID- 11379213 TI - Feminization as a critical component of the changing hydrosocial contract. AB - The Dublin Principles recognize the role that women play in water resources management. The South African Minister of Water Affairs, Prof. Kader Asmal, coined a new expression by referring to the "feminization of water". The article explores some of the ramifications of this and shows that the two aspects are not necessarily the same thing. Feminization does not necessarily mean bringing more women into management processes as it is often depicted. This is the quantitative aspect that is often referred to by male managers and it has been given a negative implication as a result. The more important issue is the qualitative aspect that involves processes such as stakeholder participation, viewing alternatives before a decision is made and accountability. PMID- 11379214 TI - Workshop 6 (synthesis): making water everybody's business--the educational challenge. AB - Water awareness and communication are crucial for developing water security. Better knowledge and improved insight can lead to new ways of thinking and hence more effective handling of water-related problems. The outcomes of the workshop are summarised. PMID- 11379215 TI - Stimulating grassroot creativity for quality of life and quality of environment. AB - This paper discusses an experiment on creativity development, a systematic endeavour aimed at empowering teachers and school principals through enhancement of their capacities to introduce and manage change at their schools. It focuses on enabling them to create learning processes and climates that stimulate creativity, especially creativity that enables students to self-help and to care for the environment. The experiment uses universities as 'beach-heads'. A group of university lecturers interested to join the program is functioning as trainer, consultant, motivator, network builder, partner, and facilitator for the teachers. The undertaking applies a multifaceted approach, involving the creation of a sense of urgency to change, restoring self-respect of the teachers. It also includes ways of starting the change from oneself, encouraging small but many practical meaningful activities, process oriented, recognizing efforts, collaborative network, cascading expansion, snowballing effects, self-selection and creating common ownership. This experiment had developed many self-organizing learning communities, inter-related and mutually benefiting in a national network for creativity. The teachers' involvement in creativity activities had brought changes in their attitudes toward students, their profession, environment, and toward change itself. PMID- 11379216 TI - How water scarcity will shape the new century. AB - Water resources are increasingly being overexploited, such that current food production, which relies heavily on irrigation schemes, is unsustainable. Many steps, including improved irrigation techniques, more water-efficient crops and animal protein production, etc., will be needed to raise water productivity across the board. Water must in future be recognised as a scarce resource and not taken for granted. PMID- 11379217 TI - Workshop 7 (synthesis): towards a recycling society: systems approach to small scale reuse of human waste. AB - Population increase and growing quantities of human excreta create serious problems and high risks for people's health. Alternative solutions are becoming crucial to improve urban living conditions, and to shorten water and nutrient flows into circulation of used water and nutrient in human excreta. The speakers presented a wide range of experiences of "closing the loops" and thereby turning potential waste into productive use. The focus of the workshop deliberations was on simplifying the hygienisation of urine and faeces and the reuse in food production by using urine-diverting toilets, as well as innovative ways to recycle sewage after various stages of treatment. PMID- 11379218 TI - Towards a recycling society: ecological sanitation--closing the loop to food security. AB - A recycling society must switch from linear solutions to circular approaches, protecting ecosystems and harmonising with natural systems. Ecological sanitation, an alternative to conventional approaches, considers excreta a resource. Excreta are rendered safe at the source prior to reuse. The ecosystem approach helps restore soil fertility, and ultimately enhances food security. It is a system requiring little or no water; thus it conserves and protects fresh and marine water sources, enhances biodiversity, and it may confer multiple benefits in urban settings. It can generate jobs, be financially more affordable than conventional approaches, improve local ecology, create decentralised economies, and contribute toward improved health and nutrition of people. Ecological sanitation closes water and nutrient loops, reducing reliance on external inputs and reducing output of wastes from the system. PMID- 11379219 TI - Workshop 8 (synthesis): simple, small-scale and appropriate technology for potable water. AB - The low quality and quantity of water supply in much of the world is best tackled by a multidisciplinary, small-scale, appropriate-technologies approach. PMID- 11379220 TI - Photobiological effects of polychromatic medium pressure UV lamps. AB - Ultraviolet (UV) light has become widely accepted for the disinfection of potable water, process water and wastewater as an alternative to chlorination. To avoid the failure of a UV disinfection system due to the recovery of micro-organisms, certain additional wavelengths in the UV area are emitted by newly developed UV lamps. To reduce the chance of microbial recovery after ultraviolet irradiation, damage must be inflicted in as many areas of the micro-organism as possible. The effective killing of micro-organisms by improved polychromatic medium pressure UV lamps is due to their exceptionally high UV energy output at specific wavelengths across a broad section of the UV spectrum. The combination of these properties results in several different lethal effects in small and large micro-organisms. Important biological molecules other than DNA are likely to be damaged, which helps to prevent the recovery of irradiated micro-organisms. Absorption line spectra of absorbing nucleotide bases, DNA and other biological molecules, including proteins and enzymes, show how effective UV light can be. Recent findings on the biological effects of short wavelengths on Bacillus subtilis, Cryptosporidium parvum and Escherichia coli confirm the effect of short wavelengths. Practical comparisons with conventional low pressure UV lamps at equal UV dosages show better killing rates from polychromatic medium pressure lamps, without formation of disinfection byproducts (DBPs). PMID- 11379221 TI - Hydrosolidarity intergenerational challenges: long-term commitment for long-term issues. AB - The year 2000 Young Professionals Seminar focused on long-term intergenerational challenges. Water related problems are symptoms of complex and ultimately societal problems linked to human behaviour, political support and managerial and institutional structures. Although integrated water resources management is presented as a solution, it is not always well understood, and can create a sense of hopelessness among professionals. To make it operational requires long-term commitments among various professionals and the involvement of new actors. A number of key topics crystallised as needing further attention, including ethical dimensions in policy making, the development of a framework for a "Future Generation Impact Assessment" (FGIA), and efforts to achieve true dialogue among stakeholders. Young water professionals must become more involved in political processes and take active part in institutional changes. Such engagement will require changes in the working environment facing many young professionals that causes frustration due to inefficient and conservative hierarchical structures and the lack of transparency. PMID- 11379222 TI - Water is a catalyst for peace. AB - Despite much recent rhetoric predicting growing conflict, even wars, over water, the weight of historical evidence points overwhelmingly to peaceful resolution of such conflicts. Water in fact tends to act as a catalyst for cooperation. PMID- 11379223 TI - From vision to action after the second world water forum. AB - The World Commission for Water in the 21st Century set out the vision of a "water secure world" and suggested a comprehensive set of measures to achieve this aim. Criticism of the role of water pricing in these measures underestimates the strength of its case in environmental, financial, economic, social, political and ethical terms. The Framework For Action has been devised as a guide for local action, but the proposal for a Water Innovation Fund may have particular impact. PMID- 11379224 TI - Ecology and equity: key determinants of sustainable water security. AB - Trends in water consumption indicate that demand for water for household and industrial uses in developing countries could double as a proportion of total water demand in the next 25 years. Scope for expansion of water supply will, at the same time, be limited because development of irrigation and urban water supplies is becoming increasingly expensive, and often involves high costs in terms of environmental degradation and human resettlement. Without fundamental reform of water management, the rapid growth in urban water demand will require large transfers of water from irrigated agriculture, thereby threatening food security. Hence, water supply and demand should be managed in an integrated fashion, simultaneously considering all uses and sources. This will call for the establishment of community centred food and water security systems and national water trusts. Once such systems and Trusts are established there could be a legally binding Global Water Convention on the model of the Global Convention on Climate and Biodiversity. The details of such a Global Water Conventions can be finalized at one of the future Stockholm Water Symposia. There are uncommon opportunities today for a water-secure world through synergy between technology, public policy and peoples' participation. PMID- 11379225 TI - Missing and neglected links in water management. AB - In the current revolution in water management; issues that must be addressed include both urbanisation and ruralisation, water quality, and globalisation and energy policy. Water management must struggle against inappropriate research, myths and inadequate data. PMID- 11379226 TI - The value and price of water (the women of Lutsheko). AB - The recommendations in the World Water Vision for full cost pricing fail to comprehend the reality of deep poverty. For the very poor, particularly in rural areas, systems that provide subsidies simply to be paid back in water charges are neither appropriate nor workable. Nor does charging for water that supplies people's basic needs have a significant impact (when compared to large users in agriculture, industry etc) on resources management. The related issue of private sector involvement is also shown to need care and caution to achieve successful results. PMID- 11379227 TI - The critical role of water in sustainable growth. AB - Business can play an important role in protecting and improving water quality by: raising public awareness; reducing and recycling water within manufacturing operations; reducing the use of/impact on water in the use of products; forming partnerships with private sector organizations focused on pilot programs to improve water quality; and by integrating water as a critical and strategic issue in all business strategies. PMID- 11379228 TI - Water scarcity and food security: alternative futures for the 21st century. AB - Water availability for agriculture--the major water user worldwide--is one of the most critical factors for food security in many regions of the world. The role of water withdrawals in irrigated agriculture and food security has been receiving substantial attention in recent years. This paper addresses key questions regarding water availability and food security, including: How will water availability and water demand evolve over the next three decades, taking into account availability and variability in water resources, the water supply infrastructure, and irrigation and nonagricultural water demands? What are the relationships among water scarcity, food production, and food security? How much of future food production will come from rainfed and irrigated areas? A global modeling framework, IMPACT-Water, is applied to explore answers to these questions using analysis. PMID- 11379229 TI - Green water security for the food makers of tomorrow: windows of opportunity in drought-prone savannahs. AB - The largest remaining biophysical water challenge is whether there is enough fresh water to sustain global food production and service natural ecosytems. Focussing on the drought-prone savannahs and small-scale farming, this paper argues that the crucial resource is vapour flow, not "blue" water, and there are grounds for optimism if resource management can be improved. PMID- 11379230 TI - Commodification of water--the wrong prescription. AB - The world faces a crisis of water stress and scarcity. This paper argues that current policies of economic globalization and privatisation are exacerbating environmental problems and that the real solutions will have a fundamentally different ethical basis. PMID- 11379231 TI - Salinization: unplumbed salt in a parched landscape. AB - The global hydrological and salt cycles are described, as are the ways in which human activities have led to their disturbance. One effect of this disturbance is the unnatural increase in the salinity of many inland waters (secondary salinization). The geographical extent of secondary salinization is outlined, together with its effects on various types of inland waters, such as salt lakes, freshwater lakes and wetlands, and rivers and streams. The likely impact on salinization of global climate change is summarized. PMID- 11379232 TI - Sustainable urban water systems in rich and poor cities--steps towards a new approach. AB - The 'big pipes in, big pipes out' approach to urban water management was developed in the 19th century for a particular linear urban form. Large, sprawling car-dependent cities are pushing this approach to new limits in rich cities and it has never worked in poor cities. An alternative which uses new small-scale technology and is more community-based, is suggested for both rich and poor countries. The Sydney Olympics and a demonstration project in Java show that the approach can work. PMID- 11379233 TI - [Mammography. Screening or not]. PMID- 11379234 TI - [Respecting the donor blood]. PMID- 11379235 TI - [Adverse effects should be reported]. PMID- 11379236 TI - [New discoveries about the cause of diabetes. Type 2 diabetes mellitus changed to "obesity-dependent diabetes mellitus"]. PMID- 11379237 TI - [Nerve affection in connection with bloodletting I. A rare, but potentially serious complication]. AB - Based on clinical experiences and a review of the literature we describe venepuncture of blood donors complicated by an affection of the peripheral nerves secondary to an arterial or venous lesion. Such complications are rare but potentially disabling and should be known in blood banks and clinical departments to which these patients are referred. Blood donors should be offered optimal and early diagnostics, which appear to be a prerequisite for treatment of donors complaining of pain of neuropathic character extending beyond the puncture site. Recognition of the complication opens options for prevention. PMID- 11379238 TI - [Nerve affection in connection with bloodletting II. Four case reports]. AB - Four blood donors with persistent pain and dysfunction of the upper extremity after venepuncture are presented. Physical examinations identified median nerve affections in all and additionally a varying involvement of other nerves in the elbow region. Such complications appear to have developed following to a vascular lesion. They are rare but potentially disabling and should be known in blood banks and clinical departments to which these patients are referred. PMID- 11379239 TI - [Jet leg]. AB - The aim of the paper is to give a review of jet-lag (Time Zone Change Syndrome) with regard to aetiology, symptomatology and pathophysiology. Furthermore we give recommendations on treatment of jet-lag. The literature has focused intensely on the potential benefit of phototherapy and/or use of melatonin as treatment modalities for jet-lag. Both phototherapy and melatonin have the capability to accelerate reentrainment of the circadian rhythm after flights across multiple time zones, thereby reducing jet-lag. We stress the importance of correct timing of phototherapy and use of melatonin and suggest that the traveller adapts to the "social" rhythm at the flight destination as well. PMID- 11379240 TI - [Referrals to clinical mammography in a county without screening program]. AB - Systematic mammography-screening may reduce the number of unindicated diagnostic mammographies. Six thousand, eight hundred and four women from Rosklde County underwent mammography at the Roskilde County Hospital over a four-year-period. Five hundred and nineteen women were excluded from the study, leaving 6285 women to form the study population. Twenty-four percent of the women had no clinical indication for mammography, 15% of the women had an increased risk of breast cancer, and for 61% of the women there was a clinical indication for mammography. Breast cancer was found in 0.46% of the women undergoing mammography without a clinical indication, in 1.2% of the women belonging to risk-groups, and in 8.9% of the women with a clinical indication for mammography. Biennial screening for breast cancer of 15,394 women (7697 women a year) in the age group 50-69 years could reduce the number of diagnostic mammographies by 338 women a year. Cooperation with GPs in a reduction of unindicated clinical mammographies of women younger than 50 years could reduce the number by a further 171 women. Potential savings in the X-ray-departments is not a weighty argument in favour of introduction of mammography screening. PMID- 11379241 TI - [Reconstruction of upper esophagus after treatment of cancer in the larynx, hypopharynx and upper esophagus]. AB - INTRODUCTION: Two new methods of reconstructing the proximal oesophagus after resection of cancer in the larynx, hypopharynx, or proximal oesophagus are examined. MATERIAL AND METHOD: Between 1991 and 1996, 12 patients were operated on after initial irradiation. The reconstructions were done by a microsurgical technique, with eight jejunum transplants and four tubulated radial forearm flaps. The records were reviewed retrospectively. RESULTS: The patients were hospitalised for about a month and were able to swallow soft food three weeks after the operation. Most patients achieved pain palliation and a good swallowing function for a period of three to more than 24 months, until relapse. All patients except two died between five and 21 months after the operation. DISCUSSION AND CONCLUSION: Both techniques give an acceptable swallowing function until relapse. The size of the defect indicates the choice of flap. They are also suitable in such situations as complex fistula formation, congenital atresia, and the after-effects of corrosion. PMID- 11379242 TI - [Antibiotic treatment of infections in general practice. Effect of audit assessed by prescriptions data from health insurance registry and physicians' own registration]. AB - The aim was to study whether an audit of treatment of infections in general practice resulted in changed prescribing habits. In 1995-1996 forty-six general practitioners (GP's) from the County of Roskilde participated in an audit regarding infectious diseases (incl. course participation and preparation of treatment guidelines). The effect evaluation was done on the basis of 1) two self registrations of antibiotic prescriptions carried out with one year's interval, and 2) prescribing data from the National Insurance database collected over two periods, before the first and second self-registration respectively. The number of patients not receiving antibiotics increased significantly from 47.2% to 52.4% after intervention. The self-registration did not show any change in choice of antibiotics, while the registry data showed a shift from broad-spectrum to narrow spectrum penicillin. This change was, however, also found among the GP's, who did not participate in the audit. The study demonstrated that audit can result in changes in prescribing patterns, but at the same time emphasizes the need for inclusion of external data sources and control groups in the evaluation of intervention effects. PMID- 11379243 TI - [Infantile botulism caused by honey]. AB - We report a case of a 5-week-old infant admitted with respiratory arrest. He had been fed with honey for two weeks. Infant botulism was suspected and confirmed by the finding of Clostridium botulinum toxin in the serum and faeces, and in the honey. The infant needed 7.5 months of ventilatory support. PMID- 11379244 TI - [Buschke-Ollendorff syndrome. Connective tissue nevi in osteopoikilosis]. AB - We describe here a patient with Buschke-Ollendorff syndrome (BOS) and present a review of the literature. BOS is characterised by osteopoikilosis, small sclerotic changes in the bones, and by dermatofibrosis lenticularis disseminata, yellow-white nodules in the skin. It is a benign, hereditary condition, with no influence on the patient's health. Awareness of this condition is of importance, as the wrong diagnosis can lead to unnecessary worry for the patient and extensive costs for the health care system. PMID- 11379245 TI - [New European guidelines for urinalysis]. PMID- 11379246 TI - [Breech delivery--Cesarean section or vaginal birth?]. PMID- 11379247 TI - [Preserve clinical neurophysiology]. PMID- 11379248 TI - [Treatment of upper respiratory tract infections]. PMID- 11379249 TI - [An awful story--and it's background]. PMID- 11379250 TI - [Is evidence-based medicine evidence-based?]. PMID- 11379251 TI - [Vigabatrin and visual fields defects]. PMID- 11379252 TI - Accuracy of prediction of walking for young stroke patients by use of the FIM. AB - BACKGROUND AND PURPOSE: Clinical prediction of walking outcome after a stroke is essential for effective discharge planning. However, its accuracy has hardly been explored. This study took place in a regional unit admitting patients with complex neurological disabilities for specialist inpatient rehabilitation. The aim was to compare predicted outcome (goal score) with achieved outcome (discharge score) on the seven-point locomotion subscale of the Functional Independence Measure (FIM), to evaluate its precision and identify factors influencing accuracy. METHOD: Admission, goal and discharge scores were analysed retrospectively for 141 subjects (90 M; 51 F) admitted consecutively to the Unit with median age 54 years (range 15-68 years) with median length of stay 13.6 weeks (range 3-35 weeks). RESULTS: Ninety subjects (64%) gained from two to six points; 50 subjects (35%) gained one point or showed no change. One patient deteriorated by two points. Excluding patients admitted with the highest score (FIM level 7), the overall level of agreement between predicted and discharge scores was moderate (weighted kappa 0.47). Prediction was accurate to +/- 1 point in 113 subjects (80%). Overprediction by > or = 2 points occurred in 16 subjects (11%) and underprediction by > or = 2 points in 12 subjects (9%). Analysis of the most-disabled cohort, admitted with FIM levels 1 or 2 scores, revealed a higher sensitivity for predicting 'independence' (FIM levels 5-7) (78%) than 'dependence' (FIM levels 1-4) (65%). Accuracy was not affected by age, gender or side of stroke. Inaccurate predictions were associated with lower admission FIM level scores (p = -0.26; p = 0.002) and a greater length of stay (p = 0.36; p < 0.001). Subjects with quadriplegia were more likely to have inaccurate outcome predictions made than those with hemiplegia (p = 0.025) and those with neglect were more likely to have inaccurate outcome predictions made than those without neglect (p = 0.017). CONCLUSION: Further investigation into clinical prediction and the variables which confound accuracy is needed for effective planning. PMID- 11379253 TI - Intra- and inter-tester reliability and reference values for isometric neck strength. AB - BACKGROUND AND PURPOSE: Age- and sex-specific reference values for neck strength based on reliable measurements in the upright position are lacking. The aim of the present study was to determine intra- and inter-tester reliability and age- and sex-specific reference values for isometric neck strength in extension, flexion and lateral flexion in sitting position measured with the David Back Clinic 140 (DBC 140) equipment. METHOD: The reliability of the DBC 140 equipment was investigated in 30 healthy volunteers and reference values were obtained from 101 healthy men and women. RESULTS: The reliability study showed that neck strength measured with the DBC 140 equipment has almost perfect intra- and inter tester reliability (ICC values between 0.85 and 0.97). The mean value of the first in a series of three measurements was the highest for all three test leaders and for almost all directions. Results from the reference value study showed that gender is a much more important determinant of neck strength than age, body weight or body mass index (BMI). Neck strength in women was, on average, 55% of that in men, and when adjusted for body weight or BMI, the percentages were 70% and 59%, respectively. In all directions observed, neck strength decreased by approximately 20% from age 25 to 64 years. CONCLUSIONS: Measurements of neck strength taken in upright position with the DBC 140 equipment have almost perfect intra- and inter-tester reliability and justify the use of this test procedure. The use of the first measurement in a test series can be recommended for use in clinical practice since it was shown to be the maximal test value and thus, had a very low intra-tester difference. The use of reference values for neck strength when evaluating patients with neck disorders needs to take gender into account. PMID- 11379254 TI - Exercises and education as secondary prevention for recurrent low back pain. AB - BACKGROUND AND PURPOSE: Exercise and education is a common physiotherapy approach in the prevention of low back pain. A Mensendieck exercise programme consisting of exercises and ergonomical education has, in a previous study, been shown to be effective in preventing recurrent low back pain during one-year follow-up. The purpose of the present study was to evaluate the long-term effect of the Mensendieck exercise programme on people with recurrent episodes of low back pain who, when entering the study, had finished treatment for their last episode of low back pain. METHOD: A randomized controlled clinical trial in which 41 women and 36 men were allocated to either a Mensendieck or control group. The Mensendieck subjects received 20 group sessions of exercises and ergonomical education over 13 weeks. The control subjects were not offered any prophylactic therapy, but were free to receive treatment or exercises. Outcome measures were the number of recurrences of low back pain, sick leave, low back function and general functional status. RESULTS: At three-year follow-up, 11 subjects had been lost to the study. Survival analysis showed a significant reduction (p = 0.02) in subjects experiencing recurrent low back pain in the Mensendieck group compared to the control group. Significant improvements in pain and function scores were reported in both groups. There was no significant difference between the groups in pain, function or sick leave. CONCLUSION: A Mensendieck exercise programme seems efficient in reducing recurrent episodes of low back pain at three-year follow-up, but it did not influence sick leave, pain or function scores. PMID- 11379255 TI - Clinical reasoning of an experienced physiotherapist: insight into clinician decision-making regarding low back pain. AB - BACKGROUND AND PURPOSE: Investigation of the clinical reasoning skills of experienced clinicians provides insight into decision-making in the practice of physiotherapy. The purpose of the present study was to analyse the clinical reasoning skills of an experienced physiotherapist during her assessment and treatment of clients with low back pain. METHOD: Deconstruction of the physiotherapist's reasoning process was accomplished through observation of encounters between her and each of six patient subjects. Reconstruction and analysis of the physiotherapist's decision-making process was performed through retrospective interviews and reflective analysis of her clinical reasoning during each encounter. RESULTS: A working model of the physiotherapist's clinical reasoning was created from an integration of theoretical elements in the literature and the data. Through analysis of this framework, two core dimensions of her clinical reasoning were revealed: the influence of clinical experience and the influence of advanced training in a specific philosophy of treating the spine. CONCLUSIONS: The construction of these themes has contributed to the growing understanding of clinical reasoning strategies and skills used in orthopaedic physical therapy practice. Detailed description of the physiotherapist's reasoning process provides more meaningful understanding of physiotherapy treatments. In this case the physiotherapist employed a pattern recognition strategy and forward reasoning process in making a diagnosis. Further research is necessary to expand knowledge on the development of clinical reasoning skills. PMID- 11379256 TI - A 12-year follow-up of subjects initially sicklisted with neck/shoulder or low back diagnoses. AB - BACKGROUND AND PURPOSE: Neck/shoulder and low back pain are common in the Western world and can cause great personal and economic consequences, but so far there are few long term follow-up studies of the consequences of back pain, especially studies that separate the location of back pain. More knowledge is needed about different patterns of risk factors and prognoses for neck/shoulder and low back pain, respectively, and they should not be treated as similar conditions. The aim of the present study was to investigate possible long-term differences in neck/shoulder and low back symptoms, experienced over a 12-year period, with regard to work status, present health, discomfort and influence on daily activities. METHOD: A retrospective cohort study of individuals sicklisted with neck/shoulder or low back diagnoses 12 years ago was undertaken. Included were all 213 people who, in 1985, lived in the municipality of Linkoping, Sweden, were aged 25-34 years and who had taken at least one new period of sickleave lasting > 28 days with a neck/shoulder or low back diagnosis. In 1996, a questionnaire was mailed to the 204 people who were still resident in Sweden (response rate 73%). RESULTS: Those initially absent with neck/shoulder diagnoses rated their present state of discomfort as worse than those sicklisted with low back diagnoses. Only 4% of the neck/shoulder group reported no present discomfort compared with 25% of the low back group. Notably, both groups reported the same duration of low back discomfort during the last year, which may indicate a higher risk for symptoms in more than one location for subjects with neck/shoulder problems. CONCLUSIONS: Individuals with sickness absence of more than 28 days with neck/shoulder or low back diagnoses appear to be at high risk of developing long-standing symptoms significantly more so for those initially having neck/shoulder diagnoses. PMID- 11379257 TI - Do physiotherapists have a role in palliative care? PMID- 11379258 TI - [Carriers of chronic hepatitis B]. PMID- 11379259 TI - [The chronic Philadelphia chromosome-negative myeloproliferative syndrome I. Pathogenesis and pathophysiology]. AB - In polycythaemia vera (PV) the erythroid progenitors proliferate autonomously independently of the circulating erythropoietin. The progenitors are hypersensitive to various growth factors, including insulin-like growth factor 1, which inhibits apoptosis in erythroid and myeloid progenitor cells. No change has been found in the erythropoietin (EPO) receptor. Thrombopoietin (Tpo) regulates the production of haematopoietic progenitor cells, particularly of platelets. By inhibiting apoptosis, this growth factor may be responsible for the autonomous proliferation of the megakaryocyte cell lineage in PV and idiopathic myelofibrosis (IMF), which are featured by highly elevated circulating Tpo levels. Thrombopoietin may also be involved in the pathogenesis of myelofibrosis and development of extramedullary haematopoiesis. Both fibrogenesis and angiogenesis in the bone marrow, spleen, and liver develop secondary to the release of various growth-promoting factors from the megakaryocyte cell lineage. The lesion of the pluripotent stem cell in PV and IMF seems to imply several defects, including lack of or decreased expression of the Tpo receptor, alterations in the sensitivity of progenitor cells to various growth factors, and alterations in important gene systems (Bcl-2), which govern cell survival. Essential thrombocytosis seems to be a heterogeneous disease entity, as about 50% of the patients have polyclonal haematopoiesis. PMID- 11379260 TI - [The chronic Philadelphia chromosome-negative myeloproliferative syndrome II. Clinical findings, diagnostics and treatment]. AB - Polycythaemia vera (PV) and essential thrombocytosis (ET) are clinically characterised by non-specific neurologic symptoms, peripheral circulatory disturbances (acrocyanosis, wounds, erythromelalgia) or abdominal symptoms. The treatment of PV includes phlebotomy, antiaggregation and cytoreduction. In ET, the primary treatment is also low-dose aspirin except for patients presenting with a haemorrhagic diathesis. Hydroxyurea may be associated with an increased risk of acute leukaemia or myelodysplasia. Therefore alpha-interferon and anagrelide should be considered in younger patients. Early cytoreductive therapy is advocated in patients with idiopathic myelofibrosis (IMF) to inhibit further progression of bone marrow fibrosis and further expansion of myeloid metaplasia in the spleen and liver. Treatment with androgens (danazol) and glucocorticoids may improve severe anaemia and thrombocytopenia. In younger patients, allogeneic bone marrow transplantation should be considered. PMID- 11379261 TI - [Acute thrombotic complication at the debute of the Philadelphia chromosome negative myeloproliferative syndrome]. AB - METHODS: We describe eight patients with a diagnosis of a chronic myeloproliferative disorder, characterised in most patients by severe thrombotic complications at the debut of the disease. RESULTS: The symptoms were life threatening in seven patients: acute upper gastrointestinal haemorrhage from oesophageal varices in four, an acute abdominal catastrophy owing to mesenteric vein thrombosis with intestinal gangrene in two, and a large cerebral infarction, which was lethal, in one. The same patient also suffered a thrombosis of the axillary and subclavian veins. Neurological symptoms, with headache, visual disturbances, dizziness, and impaired memory, were initial cardinal symptoms. In two patients, explorative laparotomy was performed with intestinal resection owing to gangrene. One patient had a toe amputation. DISCUSSION: The above symptoms are explained by thrombosis in the microcirculation because of thrombocytosis and circulating platelet aggregates. In patients with polycythaemia vera, the elevated haematocrit contributes significantly to the impaired microcirculation. Early diagnosis and management of these disorders are of utmost importance to prevent the potentially life-threatening complications described above. PMID- 11379262 TI - [Hepatitis B screening of pregnant women and follow-up vaccination of infants with HBsAG positive mothers. A survey from the Hvidovre hospital in 1999]. AB - The aim of this study was to assess the implementation of the recommendations of the Danish National Health Service concerning screening of women from hepatitis B (HB) endemic areas and follow-up vaccination of infants of carrier mothers. We examined all birth notification (n = 836) at Hvidovre Hospital during the first three months of 1999. Among 188 women from HB endemic areas only 134 (71%) women had been screened for HBsAg before delivery. Six women were HBsAg positive one of whom was examined after the delivery. Four out of six infants (67%) received Aunativ and Engerix-B within 12 hours of birth, one infant received Aunativ and Engerix-B at four days of age, the infant born of the mother who was screened after delivery didn't receive either Aunativ or Engerix-B. Three out of the six infants (50%) had received all the scheduled Engerix-B vaccinations. Our results suggest it may be necessary to reconsider the current screening and vaccination procedures to prevent an increase in the number of children becoming chronic HB carriers in Denmark. PMID- 11379263 TI - [Forensic examination of sexually abused children]. AB - INTRODUCTION: The Department of Forensic Medicine, Aarhus, performs examinations of children suspected to have been sexually abused when reported to and requested by the police. A preliminary study was taken to evaluate all cases in one year including the legal outcome. MATERIAL AND METHODS: The material included all cases in one year, all examined on request by the police. All written material including court decisions were reviewed. RESULTS: The material included 34 cases with three boys, median-age 11 years, and 31 girls, median age six years, at the time of examination. The sexual abuse events were fondling, vaginal (14), anal (7) and oral (5) intercourse as well as showing pornography. The medical examination was most often performed more than one week after the abuse. The examination revealed normal findings in 20 cases, non specific findings including erythema of the vestibulum in 13 cases and in only one child was a traumatic lesion with rupture of the hymen seen. The perpetrators were above 25 years of age and were family members or someone known to the child. Eight perpetrators were convicted in court, of whom three admitted having abused the child. DISCUSSION: The time interval between the sexual abuse and the time of examination is important to the ano-genital findings at the examination. A medical examination in cases of sexual child abuse seldom provides legal proof of sexual abuse. The most important evidence is the story told by the child. Therefore, the examination is a supplement which may support or remain neutral to the story told by the child. PMID- 11379264 TI - [Peroperative "lavage" and primary anastomosis in acute left-sided colon ileus]. AB - AIM: A description of technique, course and results of operation for left-sided obstruction of the colon with primary resection, perioperative lavage of the colon and primary anastomosis. DESIGN: Retrospective examination of case records. MATERIAL: From January 1st 1994-December 31st 1997 eight operations were performed including primary resection, perioperative lavage and primary anastomosis. RESULTS: No anastomotic leaks were found. One patient died five days postoperatively due to cardiac complications. Three patients had postoperative cardiopulmonary complications treated medically. One patient had rupture of the fascia, was reoperated, experienced wound infection, transient serum elevation of liver enzymes and underwent ERCP with papillotomy and extraction of stones from the common bile duct two months postoperatively. CONCLUSION: In this small series the technique has appeared safe, thus confirming previous reports. PMID- 11379265 TI - [Recurring epileptic seizures--a possible side effect of transcutaneous electric nerve stimulation]. AB - A case of repetitive tonic/clonic epileptic fits in a post ischaemic stroke patient after transcutaneous electrical nerve stimulation (TENS) is described. The author concludes that it is not unlikely that the TENS treatment may have contributed to the fits. PMID- 11379266 TI - [WHO consultation on the increasing incidence of campylobacteriosis in humans. Copenhagen, 21-25 November 2000, in cooperation with the National Veterinary Laboratory and the National Serum Institute]. PMID- 11379267 TI - [Dimension of the Danish scientific medical research?]. PMID- 11379268 TI - [The eight's decade]. PMID- 11379269 TI - [Preimplantation vs prenatal diagnostics]. PMID- 11379270 TI - Trigeminal autonomic cephalgias (TACs). AB - The concept of a group of headaches whose pathophysiological focus revolves around the trigeminal-autonomic reflex fills a useful gap in characterising a number of primary headache syndromes. Broadly, these syndromes involve activation of trigeminovascular nociceptive pathways with reflex cranial autonomic activation. Clinically, this physiology predicts pain with some combination of lacrimation, conjunctival injection, nasal congestion, or eyelid oedema. Several of the primary neurovascular headaches, notably cluster headache, paroxysmal hemicrania and short-lasting neuralgiform pain with conjunctival injection and tearing (SUNCT), seem to immediately fit this classification. This physiology also explains why some patients with migraine present cranial autonomic features, and the concept is thus broadly useful for clinicians seeking a pathophysiological understanding of the primary neurovascular headaches. Given the known pathophysiology one can place the various treatments aimed at preventing these headaches or indeed treating the acute attacks, into context. PMID- 11379271 TI - Trigeminal neuralgia. Pathophysiology and treatment. AB - Trigeminal neuralgia is a very peculiar disease. The pain, also known as "tic douloureux", is paroxystic and very severe. It can be triggered by a light cutaneous stimulus on a very localized spot on the face (the so-called "trigger zone"). The patient can sometimes benefit from long remissions without any treatment. With the exception of multiple sclerosis and of uncommon cases of posterior fossa tumours or other lesions impinging on the trigeminal nerve, ganglion or root, trigeminal neuralgia is considered as "idiopathic". Some benign abnormality had for long been suspected. The current opinion is now in favour of a "neurovascular conflict": an artery, most often a loop of the superior or anteroinferior cerebellar artery, has an offending contact with the trigeminal nerve root, which results in localized demyelination and ectopic triggering of neuronal discharges. This hypothesis is in agreement with the relief provided by antiepileptic drugs and is supported by recent neuroimaging data. Therapeutic options are reviewed: very efficient drugs are available but fail to provide a significant relief and/or have important side effects in many cases. Surgical alternatives are available, for which guidelines are proposed. PMID- 11379272 TI - Temporomandibular disorders as a source of orofacial pain. AB - Since decades, the dental profession has been challenged by pain and dysfunction of the masticatory system. In recent years, a better understanding of the pathophysiological mechanisms of masticatory pain paralleled the increase of basic and clinical research focusing on pain in general. Consequently, so-called diagnostic techniques and treatment procedures, based upon hypothetical, sometimes dogmatic, etiological mechanisms, are increasingly questioned, and the ill-supported thoughts are gradually replaced by research-based insights. In addition, a better communication between basic scientists, researchers, and medical or dental practitioners focusing on musculoskeletal pain, has led to an improved quality of research on pain and dysfunction of the masticatory muscles and the temporomandibular joint (TMJ). This paper will try to review this progress, comment on the clinical implications, and give some suggestions for future research. PMID- 11379273 TI - Facial pain: from animal models to functional neuroimaging studies. AB - This paper summarizes some recent findings on the physiopathology of facial pain. Over the past decade, a number of animal models of facial pain have been developed. Two of these models are discussed in more detail. The model developed by Strassman and Burstein has provided a useful tool for the study of the mechanisms underlying migraine. Single unit recordings revealed that medullary dorsal horn neurons show a long-lasting increased response to dural and cutaneous periorbital mechanical and thermal stimulation after application of inflammatory agents to the dura. In addition, dural and cutaneous receptive fields largely expanded and spontaneous ongoing activity developed. These findings suggest that the extracranial hypersensitivity that is often observed in headache may have an intracranial origin. The second model that is discussed is the model of facial neuropathic pain after infraorbital nerve ligation developed by Vos and colleagues. In contrast to the previous model which is an acute electrophysiological preparation, the infraorbital nerve model is a behavioural model. It can therefore be used both for electrophysiological and behavioural studies. In recent years, a number of functional neuroimaging studies of facial pain appeared. Studies performed in cluster headache patients seem to point to a crucial role of the hypothalamus in this syndrome. However, since hypothalamic activation has also been reported in some other types of (facial) pain, the specificity of the role of the hypothalamus in cluster headache remains to be proven. PMID- 11379274 TI - Type A botulinum toxin in the treatment of chronic facial pain associated with temporo-mandibular dysfunction. AB - In an open-label study 41 patients suffering from the muscular form of temporo mandibular dysfunction were treated with botulinum toxin type A injections into masticatory muscles (average of 200 U on each side) and followed for an average of 6.7 months. Eighty percent of patients improved by a mean reduction of 45% on a visual analogue pain scale. During the observation period, 17% of patients had to receive a second injection because of recurrent pain. Reversible speech and swallowing difficulties occurred in only 1 patient. These encouraging results need to be confirmed by a randomized controlled trial. PMID- 11379275 TI - New generation anti-epileptics for facial pain and headache. AB - The prophylactic management of recurrent head and facial pains may be challenging because of lack of efficacy and/or bothersome adverse effects of available drug therapies. New generation antiepileptic drugs offer new perspectives in difficult cases. We will review the available published data and present our experience with lamotrigine in various head and facial pains such as migraine, cluster headache, neuropathic trigeminal pain, atypical facial pain, and chronic tension type headache. Twenty-five patients were enrolled and followed for 18 months. The dose was gradually increased in steps of 25 mg up to the effective dose (mean 250 mg/d). Lamotrigine was most effective in trigeminal neuralgia and dysesthesia, but was of little utility in the other head or facial pains. PMID- 11379276 TI - Neuropathic pain: the clinical syndrome revisited. AB - Neuropathic pains associated with an injury of the peripheral or central nervous system are among the most difficult to treat. One of the reasons for the therapeutic difficulties in these patients is that the pharmacological treatments are used in a uniform fashion whatever the clinical picture, while these syndromes are in fact highly heterogenous. The patients can express various combinations of painful symptoms--spontaneous (continuous and/or paroxysmal) and evoked (allodynia and/or hyperalgesia). Recent pharmacological studies have shown that current treatments of these pains do not induce global and uniform analgesic effects but rather act preferentially or selectively on some of their components. Such data emphasize the necessity of a thorough evaluation of patients presenting with neuropathic pains, notably by using quantitative sensory testing. Following recent advances in the understanding of the pathophysiological mechanisms underlying these painful syndromes, through experimental studies in animals, a "mechanism-based" classification and treatment of neuropathic pains can be envisaged. The main goal for clinicians is to propose new methods and strategies for identifying pathophysiological mechanisms in patients in order to validate such an approach in the clinical context. PMID- 11379277 TI - Pharmacologic treatment of neuropathic pain. AB - There is currently no consensus concerning the optimal therapeutic strategy for neuropathic pain, despite an increasing number of clinical trials demonstrating successful pain relief with several drugs. Treatments have generally been selected on the basis of evidence for efficacy in randomized placebo-controlled trials conducted in disease-based groups of patients, notably in postherpetic neuralgia and diabetic polyneuropathy. These studies plead in favour of the overall efficacy of tricyclic antidepressants, standard and newer antiepileptics, opioids, tramadol, systemic and topical local anaesthetics, and some NMDA receptor antagonists; whereas evidence for efficacy is less for selective serotonin reuptake inhibitors, antiarrhythmics (mexiletine), and capsaicin. Pharmacological tests, notably therapeutic infusions, have been proposed for predicting the effectiveness of long-term treatments, but are not routinely performed. An analysis of the various neuropathic symptoms, aimed at selecting treatments targeted at mechanisms, may ultimately help the choice of different pharmacologic agents. PMID- 11379278 TI - Differential diagnosis of facial pain. AB - We will describe the differential diagnosis of primary and secondary facial pains and present illustrative case studies. The diagnosis of facial pain needs a multidisciplinary approach if the clinical presentation is not pathognomic. While patients with acute facial pain urgently need treatment, those with chronic facial pain need at priority a correct diagnosis. PMID- 11379279 TI - Thalamic stimulation in neuropathic pain: 27 years later. AB - An overview is given of CNS mechanisms which are behind the beneficial effects of VPL-VPM thalamic stimulation in the treatment of neuropathic pain. Further research in this field is urgently needed and the recent possibility to combine Deep Brain Stimulation with positron emission tomography (PET) will certainly help to unravel the brain circuitry implicated in stimulation-produced analgesia. Brain stimulation is an artificial way to activate nervous tissue that is reversible and, when correctly applied, has few complications. The clinical results warrant a continued dissemination of brain stimulation as a treatment in well selected cases of neuropathic pain. PMID- 11379280 TI - Determination of the presence of antioxidants deriving from sage and oregano extracts added to animal fat by means of assessment of the radical scavenging capacity by photochemiluminescence analysis. AB - Herbs and their extracts with antioxidant capacity could be used directly as stabilisers of fat and indirectly as feed additives, in order to improve quality and shelf-life of meat and fat-containing food. In this work a sensitive analytical method is proposed for determination of the antioxidant activity measured by photochemiluminescence (PCL) in lard stabilised with extracts of sage (Salvia officinalis L.) or oreganum (Origanum vulgare L.). A prior step of purification of fat samples is required, in order to separate and concentrate the phenolics from lipidic substances. The method was validated by determination of recovery rate and repeatability. In addition fat samples originating from pigs fed with feed additives of Salviae folium or Origani herba were analysed to investigate the supposed antioxidative effects, that could increase the shelf life of meat products. In contrast with lard mixed with extracts of sage or oregano, back fat samples originating from pigs fed with feed additives of the same herbs didn't show a higher antioxidant activity than the control group. On the one hand it seems possible to keep perishable fat-containing food longer by an addition of an extract of sage or oregano due to their antioxidative properties, on the other hand administration of feed additives of dried herbs to pigs had no effect on quality and shelf-life of fat obtained from these animals. PMID- 11379281 TI - Antioxidant activity of sage extracts in rapeseed oil irradiated with UV-rays. AB - The antioxidant activities of sage acetone oleoresin and its methanol-water fractions were tested in rapeseed oil at 0.1% concentrations and 80 degrees C temperature after irradiation with UV-rays (exposition 0.5 h). It was determined that even little doses of UV irradiation induce oxidation process in rapeseed oil samples both with additives and without them. The antioxidant activity of a synthetic antioxidant BHT was much lower than that of the sage fractions at 80 degrees C and forced air oven conditions. This finding proves the advantage of sage extracts in comparison with BHT at high temperatures. It was also found that methanol-water fraction separated from acetone oleoresin had the highest antioxidant activity in terms of the formation of primary and secondary oxidation products. Oil sample with this additive had the highest protection factor (PF = 4.53) when the oil was not irradiated with UV rays. The data obtained showed that all sage extracts had very strong antioxidant activity (PF > 3). PMID- 11379282 TI - High summer concentrations of mercury in big perch (Perca fluviatilis L) from the Tvarminne archipelago (SW Finland) and Nato (Aland Islands) Baltic sea. AB - During high summer seasons in 1994-1999 84 big (20-42 cm total length) female perch (Perca fluviatilis L) were sampled from the waters around Tvarminne Zoological Station on the Hanko-Hango peninsula on the SW coast of Finland for mercury analysis. Additional sampling of 13 big female perch during the same season were carried out from the waters of Nato Biological Station on the Aland Islands in 1997-1998. High summer is the most important season for perch fishing and consumption not only for private families during leisure time but also for professional fishery including trade and export, processing and preparations for restaurants. Fillets of big specimen of perch, s.c. "fillet perch" or "sea perch" from outer coastal waters were chosen for the study to find out the levels of mercury concentration in these desirable fish. The mean concentration of mercury in the dorsal muscle tissue (fillet) of big female "sea perch" from the coastal waters of Tvarminne Zoological Station was calculated to 0.22 (SD 0.08), range 0.08-0.43 mg/kg, f wt and for big female perch from the waters of Nato Biological Station to 0.18 (SD 0.07), range 0.07-0.30 mg/kg, f wt respectively. The results show a considerably lower mercury level in big "fillet perch" caught from the outer coastal waters in both Finland (and Estonia) compared with the levels recorded of big "fillet perch" from fresh water lakes and reservoirs. Thus, the common expression "sea perch" may also be used in the form "Baltic sea perch" meaning big healthy perch with low concentrations of mercury and caught from the outer coastal waters in S and SW Finland (and W Estonia) when marketing perch for human consumption. PMID- 11379283 TI - Mineral absorption by albino rats as affected by some types of dietary pectins with different degrees of esterification. AB - Male albino rats were fed diets contained 6.85% mineral salts for 2 weeks (adaptation condition). Then they were fed the dietary pectin administered diet for 6 weeks to evaluate the effect of administration of pectin on the absorption of some monovalent, bivalent and heavy metals in the serum of rats. The experimental parameters included, monovalent minerals (K, Na), bivalent minerals (Zn, Cu, Ca, Fe), heavy metals (Pb, Cd), serum uric acid and serum creatinine. The obtained results indicated that the serum contents of monovalent minerals were negatively affected by pectin administration. The low degree of esterification of pectin was more effective on the absorption of bivalent minerals. Also, the rat serum levels of lead and cadmium were reduced by pectin administration. Serum total proteins were reduced by pectin administration. The level of rat serum of uric acid and creatinine fed different sources of pectin were within normal levels and were insignificantly lower than that recorded for control samples. PMID- 11379284 TI - Optimum production, stability, partial purification and inhibitory spectrum of antimicrobial compounds produced by Pediococcus pentosaceus DI. AB - The optimum conditions for production of antimicrobial compounds by Pediococcus pentosaceus DI were studied. The antimicrobial compounds in cell free fermented medium were extracted (methanol-acetone), then tested for inhibitory activity against fourteen bacterial strains and its stability to heating, various pH and proteolytic enzymes. In addition, methanol-acetone extract was partial purified through Sephadex G-100 column and thin-layer chromatography. Production of antimicrobial compounds were maximized when P. pentosaceus DI was grown in lactobacilli MRS containing 0.05% L-cysteine-HCl medium at 30 degrees C for 48 h. The methanol-acetone extract showed relatively broad antimicrobial spectrum including lactic acid bacteria and some spoilage and pathogenic bacteria. Antimicrobial activity of methanol-acetone extract was retained after heating (100 degrees C for 30 min) at pH 2.5 to 6.0 but was lost after treating with several proteolytic enzymes at pH 6.5 and 7.0. Sephadex G-100 column revealed three peaks in methanol-acetone extract. The major peak was eluted at 40 ml and including three fractions, which exhibited inhibitory activity, while the other peaks were principally free from any antimicrobial compounds. Thin-layer chromatography of crude methanol-acetone extract and the three fractions, which exhibited inhibitory activity, showed that there was one main spot with Rf at 0.8. PMID- 11379285 TI - Differentiation of Clostridium perfringens and Clostridium botulinum from non toxigenic clostridia, isolated from prepared and frozen foods by PCR-DAN based methods. AB - During the elaboration process of prepared and frozen foods, Clostridium sp. have been reported. From those microorganisms, C. perfringens and C. botulinum may pose a high risk for the consumers. To avoid these pathogenic organisms an HACCP program should be implemented, but in addition sensitive and moderately time consuming microbiological methods for monitoring C. perfringens and C. boulinum should be established. In this work, an RFLP analysis of the 16S rDNA will be developed to differentiate C. perfringens from other Clostridium sp. In addition, a PCR protocol, will be assayed to detect C. botulinum. Both two methods will be compared with biochemical characterization by API system. The restriction analysis of the 16S rDNA with Taq I and Rsa I showed at least the same sensitivity to differentiate C. perfringens from clostridial isolates as biochemical identification. However, the former method takes only 8-10 h of analysis as compared with 24-48 h required for biochemical characterization. With the specific PCR protocol to detect C. botulinum a band of 1.1 kbp was obtained derived from the specific amplification of BoNT genes, taking 6-8 h for analysis. Both two molecular DNA based methods should be considered as verification techniques of pathogenic clostridia in the HACCP program. PMID- 11379286 TI - Drying of olive pomace by a combined microwave-fan assisted convection oven. AB - The drying rate curves in terms of moisture content versus time for olive oil pomace have been experimentally obtained using microwave and combined microwave convection oven. The effects of microwave power (350, 490, 700 W), thickness (6, 9, 14 mm) and temperature (100, 160, 225 degrees C) have been investigated. It has been found that the effects of power output and thickness are negligible in microwave oven, whereas the drying rate increases with temperature increase in combined microwave-convection oven. The diffusion model fitted well to the drying curve of experimental data obtained for olive pomace at the studied conditions of drying. PMID- 11379288 TI - Quality changes during frozen storage and thawing of mixed bread. AB - In the present work investigations of the dependence between frozen storage time, the method of thawing (air blast at 50 degrees C and microwave), organoleptic and physico-chemical changes in bread are reported. The quality of the thawed product was analysed directly after thawing and after two days of storage at room temperature. It was found that changes in quality of bread are more affected by frozen storage than by the employed thawing method. The thawing methods had a significant (p < or = 0.01) effect on the investigated physico-chemical parameters of the product, however, their statistically significant (p < or = 0.01) effect on the sensory quality could only be revealed after two days of storage of the previously thawed bread at room temperature rather than directly after thawing. The results obtained in the present study suggest that bread which underwent microwave thawing had generally better quality in comparison with air blast thawing. PMID- 11379287 TI - Encapsulation of orange oil in a spray dried double emulsion. AB - Encapsulation is an important technique being used to protect sensitive food materials like flavours from deterioration. The capsule wall isolates them from the atmospheric oxygen, moisture, temperature and light. Encapsulation also masks some objectionable flavours, e.g. fish oil and some bitter antibiotics. In this study orange oil was encapsulated in the inner compartment of a double emulsion belonging to the type O1-W-O2 where O1 is orange oil, W is water and O2 is vegetable oil. In order to make orange oil double emulsion suitable for use in dry mixes, it was secondarily coated with wall materials of lactose and caseinate using spray drying technique. Entrapment of orange oil in such structure is also expected to slow down the release of volatiles and guarantee more protection for orange oil against atmospheric conditions. This method may have a potential application in different types of food or pharmaceutical products where maximum protection for flavours or slow release are required. This study includes detailed preparation of the spray dried double emulsion, evaluation of the encapsulation efficiency using light and scanning electron microscope and calculation of the yield percent of the encapsulated oil. In a separate paper we will examine the efficiency of spray dried double emulsion to control the release of orange oil by GC. PMID- 11379289 TI - Immobilization and characterization of ficin. AB - Fig tree latex (ficin) was immobilized on Celite by adsorption. The free and immobilized ficin were utilized in the production of teleme (a Turkish milk product). After immobilization, the optimal temperature of ficin was shifted from 60 to 80 degrees C. Both free and immobilized ficin exhibited higher milk clotting activity in the acidic pH range. Increase in solid content of milk affected milk clotting activity in opposite direction. When the amount of either free or immobilized enzymes doubled, clotting timereduced nearly 1.6 and 1.7 fold for free and immobilized enzyme, respectively. Use of immobilized enzyme in teleme production gave better results in terms of chemical and sensory properties compared with teleme made by free enzyme. Syneresis in teleme reduced from 70% to 60% depending on immobilization. Both acidity and bitter taste of teleme produced from free ficin was due to extensive protein hydrolysis by proteolytic fractions. PMID- 11379290 TI - Understanding the interactions of natural minor plant components with the main constituents of foods--a task for food science. PMID- 11379291 TI - Model studies on reactions of plant phenols with whey proteins. AB - Whey proteins were modified by reaction with selected phenolic compounds (ferulic , chlorogenic-, caffeic- and gallic acid) and related substances (quinic acid and p-quinone) as well as with extracts from coffee, tea, potato and pear at pH 9. The derivatives formed were characterized in terms of their physicochemical and digestion properties. The derivatization was accompanied by a reaction at the lysine and tryptophan side chains, whereby their content was decreased in comparison to that in the control whey proteins. Moreover, the solubility of the derivatives decreased over a broad pH range and the derivatization influenced the hydrophobe-hydrophile character of the whey proteins. The isoelectric points were shifted to lower pH values in the order of reactivity as follows: gallic acid > p quinone > caffeic acid > chlorogenic acid. The other derivatives showed no or few changes compared to the control whey proteins. The formation of high molecular fractions was documented with SDS-PAGE. Especially the derivatives of chlorogenic , caffeic-, gallic acid and p-quinone showed an increase in molecular weight of beta-lactoglobulin fraction from 18,300 to 20,000 Da. A dimer formation in molecular range 40,000 was also registered. MALDI-TOF-MS was applied to characterize the binding of the individual phenolic compounds or their oxidation products to the whey protein fractions, alpha-lactalbumin and beta-lactoglobulin. In vitro experiments showed that the digestion of the derivatized whey proteins with the enzymes of the gastrointestinal tract (trypsin, chymotrypsin, pepsin and pancreatin) was adversely effected. Similar results with regard to physicochemical characterization and digestion properties of the whey proteins treated with the applied extracts from plant beverages, fruit and vegetable were also documented. Coffee and tee were comparatively the most reactive extracts. PMID- 11379292 TI - Phenolics and physico-chemical characteristics of persimmon during post-harvest storage. AB - Garden picked mature but unripe fresh persimmon fruits were unipackaged in different thicknesses of polyethylene (PE) and stored at room (18.5-30 degrees C) and refrigerated temperature (6 +/- 1 degrees C). Maximum mean methanol extractable sinapine (0.168%), catechin (1.51%), and leucoanthocyanidine (10.94 delta A550/g) were recorded in the unipackaged samples kept at room temperature during 6 weeks storage, whereas the minimum values for water extractable phenolics (sinapin, catechin and procyanidin) were recorded in unipackaged samples under refrigerated temperature. The mean maximum weight loss of 12.58 and 7.90% was recorded in control samples kept at room temperature and low temperature, respectively. The weight loss for unipackaged in different thicknesses of PE ranged between 0.93-0.96% and 0.43-0.45% for samples kept at room and low temperature, respectively. Changes in texture values were significantly faster in control than unipackaged samples (P < 0.05). Low temperature combined with unipackaging in PE film performed better for the maintenance of overall fruit quality during post-harvest storage. PMID- 11379293 TI - Improvement of solubility and of emulsifying properties of milk proteins at acid pHs by esterification. AB - Three milk proteins (beta-lactoglobulin, alpha-lactalbumin and beta-casein) were esterified to different extents with methanol, ethanol and propanol, then their solubility was studied in the pH-range of 3-10. Their emulsifying properties were also checked in the pH-range of 3-7 by measuring the oil-droplet size using laser light scattering. Solubility of all esterified proteins was improved in the acidic pH range (3-6). The magnitude of improvement was depending on the extent of esterification, the nature of the grafted ester group and the nature of the modified protein. Emulsifying activity and stability of esterified milk proteins in the acidic pH range of 3-5 were generally higher compared to the corresponding native proteins. This improvement was associated with the degree of solubility, the degree of esterification, the type of ester group grafted and the nature of the used protein. The highest emulsifying activity improvement was observed for methyl ester derivatives at pH 5 where the native proteins have poor emulsifying properties. PMID- 11379294 TI - Effect of processing on the nutrients and anti-nutrients of lima bean (Phaseolus lunatus L.) flour. AB - The effect of soaking, autoclaving and toasting methods on the proximate and mineral composition and anti-nutrients of lima (butter) bean (Phaseolus lunatus L.) was investigated. The raw seed showed high content of proteins, lipids and ash with good values of K, Ca, Mg, P, Na and Fe in comparison with other legumes. Potassium was the most abundant macro mineral while sodium was the least. Iron content was the highest among the micro minerals. Manganese was not detected in any of the samples. Raw lima bean contained cyanide, trypsin inhibitor, lectin, phytin and tannin. Phytate phosphorus was calculated to be 28.2% of total phosphorus. There was no significantly different (P > 0.05) in the crude protein content of raw and processed flour. There was significant difference (P < 0.05) in the values of the macro and micro mineral. Soaking, autoclaving and toasting completely eliminated trypsin inhibitor and lectin while it significantly (P < 0.05) reduced the levels of phytin, tannin and cyanide. Except for tannin autoclaving for 20 min was found to eliminate all the other anti-nutrients in lima bean. PMID- 11379295 TI - Nutritional potential of biochemical components in Galactia longifolia Benth. (Fabaceae). AB - Seeds of Galactia longifolia Benth. (Fabaceae), locally known as Kaattukollu in Tamil language, used as food by Malayali tribals in the Kolli hills of Namakkal District, Tamil Nadu in South India were analysed for proximate composition, total (true) seed proteins, amino acid composition, fatty acid composition, minerals and antinutritional factors. Crude protein, crude fat, ash and nitrogen free extractives constitute 25.56%, 6.18%, 5.12% and 56.15%, respectively. The caloric value of 100 g dry matter of seed material is 1,601 KJ. The essential amino acids, tryptophan, leucine + isoleucine, arginine and threonine are present relatively in large quantities. The unsaturated fatty acids constitute more than 60%. The seeds are very rich in potassium, magnesium, calcium and phosphorus. Antinutritional factors such as total free phenols, tannins, L-DOPA, hydrogen cyanide and phytic acid is present in minute quantities. The seed meal is consumed after eliminating all the antinutritional factors using the conventional method of soaking seeds in water, boiling with water and decanting for four times by Malayali tribals. PMID- 11379296 TI - Locoregional therapy for liver metastases from colorectal cancer: the possibilities of intraarterial chemotherapy, and new hepatic-directed modalities. AB - Liver metastasis of colorectal cancer is a life-threatening prognostic factor. Hepatic resection, when possible, is the best therapeutic modality, although the overall survival rate is still low (30%). The diagnosis has been carried out by clinical examination, abnormal alkaline phosphatase, lactic acid dehydrogenase and tumor markers, abdominal liver echography and computed tomography scan. Angiography and intraoperative echography are useful for resection. The number of hepatic metastases and the surgical margin are probably the most significant prognostic factors. Colorectal cancer may spread predominantly to the liver making regional treatment strategies viable options. Subtotal hepatic resections and segmentectomies are potentially curable procedures for single or small numbers of hepatic metastases without other sites of disease. However, there have been no prospective randomized trials comparing patients with unresected liver metastases and resected metastases. Regional chemotherapy with floxuridine seems useful combined with hepatic resection or as palliative therapy. Gastric ulcer and biliary sclerosis are the main related toxicities. Patients with localized, unresectable hepatic metastases or concomitant bad medical condition may be candidates for radiation, percutaneous ethanol injection, cryosurgery, radiofrequency, hypoxic flow-stop perfusions with bioreductive alkylating agents, hepatic arterial ligation, embolization and chemoembolization. These new hepatic directed modalities of treatment are being investigated and may offer new approaches to providing palliation and prolonging survival. This review reports the possibilities of intraarterial chemotherapy and other novel hepatic directed approaches to the treatment of liver metastases from this common disease. PMID- 11379297 TI - Amifostine (Ethyol) as modulator of hepatic and biliary toxicity from intraarterial hepatic chemoembolization: results of a phase I study. AB - BACKGROUND/AIMS: Hepatic and biliary toxicity are still significant problems after intraarterial hepatic chemoembolization for liver metastases from large bowel cancers. In about 30-60% of the patients hepatic and biliary toxicity are the limiting aspects of intraarterial hepatic chemoembolization and exclude a lot of patients from a repeated beneficial treatment. Amifostine (Ethyol) is a prodrug that must be dephosphorylated to the free thiol in which form it can detoxify free oxygen radicals generated by radiation, hypoxia and by drugs such anthracyclines, platinum analogues and alkylating agents. Amifostine as inactive prodrug is primarily metabolized at the tissue site by membrane alkaline phosphatase, which is highly active in the cell membranes of normal endothelial cells and biliary tree cells but not in the cell membranes and neovascular capillaries of tumor. When dephosphorylated to WR-1065, amifostine is rapidly taken up into normal liver cells by a carrier-mediated facilitated diffusion transport process. The resulting high thiol content in normal liver tissue (biliary cells and hepatocytes) compared with the negligible concentration in liver metastases from large bowel cancers probably provides for selective drug resistance to intraarterial hepatic chemoembolization protecting normal tissue and allowing full therapeutic effect on tumor. METHODOLOGY: From May 1997 we planned a phase I study in patients receiving intraarterial hepatic chemoembolization for liver metastases from large bowel cancers. We started at 200 mg/m2 dissolved in 250 cc of normal saline given in 15 min in the intrahepatic artery 20 min before an intraarterial hepatic chemoembolization consisting of mitomycin 10 mg/m2, epirubicin-50, cisplatin-60 diluted in 10 mL of contrast media, mixed in 15 mL of lipiodol UF followed by a gelfoam powder solution until stagnation of the flow. The escalating dose, every 3 patients, was: 200 mg/m2, 250 mg/m2, 300 mg/m2, 350 mg/m2. RESULTS: Toxicity has been observed at 350 mg/m2: 1 patient reported transient hypotension (Blood pressure 70/50 mm Hg), 1 patient had skin flushing and dyspnoea. 300 mg/m2 are well tolerated and seem to reduce the level of transaminases, lactic acid dehydrogenase, and gamma-glutamyl transferase. Also the duration of necrotic damage, always observed after intraarterial hepatic chemoembolization, seems shorter compared with historical controls. CONCLUSIONS: Amifostine can be certainly administered at 300 mg/m2 as intraarterial infusion and could be a significant step to ameliorate the therapeutic ratio of intraarterial hepatic chemoembolization. PMID- 11379298 TI - Hepatic arterial chemotherapy for colorectal liver metastases. AB - The management of hepatic metastasis from colorectal cancer represents a significant clinical problem. For the majority of patients with colorectal liver metastases, chemotherapy will be the mainstay of treatment. Considerable data now exists to support hepatic arterial chemotherapy as being effective over systemic chemotherapy both in terms of quality of life and response rates. Morbidity and mortality rates are reduced when catheters are placed by skilled dedicated surgeons using rigorous protocols. PMID- 11379299 TI - Chronotherapy of colorectal cancer metastases. AB - Chronotherapy has consisted in the adaptation of chemotherapeutic drug delivery to circadian (approximately 24-hour) rhythms. This can be achieved in fully ambulatory patients using multichannel programmable pumps. Up to approximately 1500 patients with metastatic colorectal cancer have been registered in one of 15 trials testing the relevance of this treatment method with 5-fluorouracil +/- leucovorin +/- oxaliplatin. Chronotherapy was shown as significantly less toxic and more effective than constant rate infusion in 2 consecutive multicenter trials. High efficacy and good tolerability permitted secondary surgical resection of previously inoperable metastases, with apparent survival improvement (3-year survival > or = 20%) and cures in some patients. This strategy is currently undergoing further testing within the European Organization for Research and Treatment of Cancer. Nevertheless, combining chronotherapy with surgery of colorectal cancer metastases can be readily offered to patients as a safer therapeutic option for optimizing outcome. PMID- 11379300 TI - Cryosurgery for colorectal liver metastases. AB - Destructive therapy for liver tumors has been available for some time and a number of options have been used. These include alcohol injection, heating therapy with laser and, more recently, radiofrequency ablation and cryosurgery. The principle of cryosurgery is to destroy tissue by freezing and causing expansion of intracellular water. The effect of this treatment is to cause disruption of the cell membrane as the ice expands, thereby killing the cells and, secondly, releasing intracellular antigen into the circulation to promote an immune response. Lastly, there is a "frostbite" effect with thrombophlebitis of vessels feeding the tumor because of the freezing. A number of trials have evaluated destructive therapy, but none have shown definite benefit over chemotherapy alone. This review examines the data as it currently stands. PMID- 11379301 TI - Cryosurgery as treatment modality for colorectal liver metastases. AB - Liver metastases occur in more than 40% of the patients with colorectal cancer. The best prospect of cure is achieved by resection of the metastases. Only 10-15% of the patients with colorectal metastases are candidates for resection. When resection is impossible, other treatment options are limited. Response rates to chemotherapy are around 40%, but survival benefit is generally limited to a few months. Recently, cryosurgery has provided a new therapeutic approach for unresectable colorectal liver metastases. The metastases are localized by ultrasonography and are subsequently frozen with the aid of cryosurgical equipment. The treatment results in necrosis of the tumor and is characterized by low mortality and acceptable morbidity. Experience with cryosurgery is limited, but the results are promising. Various studies report a 1-year survival of more than 70% and a 2-year survival of > 50%. Disease-free survival after 2 years varies between 20-29%. Recurrences in the liver are usually other lesions than those treated with cryosurgery. Cryosurgery in combination with resection or as sole treatment, can result in radical treatment for patients formerly considered unresectable. Resection of liver metastases, however, remains the gold standard in the treatment of liver metastases. The main indication for cryosurgery may be as a complement to hepatic resection in those patients in whom liver resection can not achieve complete tumor clearance. PMID- 11379302 TI - Update of laser-induced thermotherapy for liver tumors. AB - Surgical resection generally offers the only chance of long-term survival for patients with primary or secondary liver tumors. Difficulties relating to the stage of the tumor, the extent of underlying liver cirrhosis and the general condition of the patient make this technique unsuitable for the majority of patients. Many nonresectional methods of in-situ tumor ablation have been recently described. These include alcohol injection, cryotherapy, radiofrequency and intraoperative radiotherapy. Recently more interest has been directed to the use of laser as a source of ablative energy. Laser-induced interstitial thermotherapy is an experimental technique used to destroy tumors within the liver. Initially this was in the form of bare-tipped photocoagulation but the development of a diffuser tip has enabled the formation of larger diameter lesions using heat to produce an area of coagulative necrosis. Less heat is lost if hepatic vascular inflow is occluded during the procedure and consequently a larger area of tumor necrosis is produced. The prospect of a simpler, more efficient system of tumor ablation is attractive so we undertook a review of the current issues surrounding this emerging treatment. PMID- 11379303 TI - Selective internal radiation therapy (SIRT) with 90Yttrium microspheres for extensive colorectal liver metastases. AB - BACKGROUND/AIMS: SIRT (selective internal radiation therapy) is a new modality for the treatment of nonresectable liver tumors which has been reported to achieve high response rates. We report our initial experience in patients with extensive colorectal liver metastases. METHODOLOGY: Thirty-eight (38) patients were treated with SIRT between February 1997 and November 1998. Liver involvement was < 25% in 19 patients, 25-50% in 9 and > 50% in 10. Patients received 90Yttrium microspheres into the hepatic artery via an arterial port and subsequent 4-weekly cycles of hepatic artery chemotherapy with 5-fluorouracil. RESULTS: SIRT was well tolerated and no treatment-related mortality was observed. Responses to SIRT as indicated by falling tumor markers and serial 3-monthly computed tomography scans were seen in over 90% of patients. Estimated survival at 6, 12 and 18 months was 70%, 46% and 46%, respectively, and was principally determined by the development of extrahepatic metastases. CONCLUSIONS: SIRT is well tolerated in patients with extensive colorectal liver metastases and achieves encouraging liver tumor responses, which are well maintained by hepatic artery chemotherapy. The modality warrants wider use and investigation. PMID- 11379304 TI - What determines survival duration in hepatocellular carcinoma treated with intraarterial Yttrium-90 microspheres? AB - BACKGROUND/AIMS: The survival duration of patients with nonresectable hepatocellular carcinoma confined to the liver and treated with intraarterial yttrium-90 microspheres was highly variable. METHODOLOGY: Eighty-two patients treated by this method were analyzed. Thirty-one patients who lived for one year or longer from the date of first treatment were classified as 'long survivors' and 51 patients who died within 1 year were classified as 'short survivors'. RESULTS: Comparison between the 2 categories suggested that lower pretreatment level of alpha-fetoprotein and higher tumor-to-normal uptake ratio of yttrium-90 microspheres favored longer survival. Results also indicated that the treatment was effective even for large tumors and for postoperative recurrence. Repeated treatment of viable residual or recurrent tumors offered further palliation and prolongation of survival. CONCLUSIONS: Pretreatment alpha-fetoprotein level, tumor-to-normal uptake ratio of yttrium-90 microspheres and the number of treatments determine survival duration. PMID- 11379305 TI - Current status of surgery in colorectal metastases to the liver. AB - The prime role of hepatic resection in the management of colorectal cancer metastatic to the liver is firmly established. The evolution of surgical and anaesthetic techniques has dramatically reduced the morbidity and mortality of elective liver resection in the hands os specialist hepatobiliary surgical teams. At least a third of patients who undergo liver resection for colorectal metastases can expect to survive five years. Although only 10-20% of all patients presenting with colorectal liver metastases are suitable for surgery, novel strategies such as portal vein embolisation and downstaging chemotherapy promise to increase the proportion of eligible patients. PMID- 11379306 TI - The argument for liver resection in colorectal liver metastases. AB - The first liver resection for colorectal metastasis was performed by Cattell in 1940. Unfortunately, there has never been a randomized control trial for liver resection against other treatment modalities (best supportive care, palliative chemotherapy, etc). Since this operation is now well established in the surgical repertoire, it would now be ethically difficult to undertake such a trial. PMID- 11379307 TI - Development of a dendritic cell (DC)-based vaccine for patients with advanced colorectal cancer. AB - BACKGROUND/AIMS: An ability to induce a specific immune response to cancer would provide an important new dimension in its management. We report our initial work investigating the safety and efficacy of a dendritic cell vaccine in patients with colorectal cancer. METHODOLOGY: Fifteen (15) patients with advanced colorectal cancer had vaccines prepared from autologous dendritic cells pulsed with tumor RNA and keyhole limpet hemocyanin. Vaccines were administered intravenously and patients were observed in hospital for 2 days. Thereafter, consultations were at monthly intervals at which time booster doses were given to a total of 4. Patients were monitored with weekly blood tests, including carcinoembryonic antigen, and 3-monthly computed tomography scans. RESULTS: Flow cytometry confirmed dendritic cell phenotype and in vitro function was confirmed by mixed lymphocyte reaction. No major adverse effects were observed. Eleven of 13 patients tested developed a positive keyhole limpet hemocyanin skin test and in 7 the carcinoembryonic antigen fell suggesting some in vivo anticancer effect. To date no dramatic clinical responses have been observed but follow-up is very short. CONCLUSIONS: The therapy was well tolerated. Dendritic cells were verified by phenotype and in vitro function. The positive keyhole limpet hemocyanin skin test confirms in vivo function by effective vaccination to keyhole limpet hemocyanin. Demonstration of any anticancer efficacy will require further follow up. PMID- 11379308 TI - Assessment of therapeutic effect of liver tumor ablation procedures. AB - Interventional procedures for percutaneous tumor ablation have gained an increasingly important role in the treatment of liver malignancies. After interventional therapies, diagnostic imaging has the key role in determining if the treated lesion is completely ablated or contains areas of residual viable neoplastic tissue. This is particularly important since in case of incomplete necrosis of the lesion, treatment can be repeated, and tumor ablation can be further pursued. The evaluation of the therapeutic effect of the procedure leads to different problems according to the histotype of the malignancy. In the case of hepatocellular carcinoma, detection of residual viable tumor is facilitated by the typical hypervascular pattern of this neoplasm. Contrast-enhanced color Doppler ultrasonography can be used to monitor tumor response, and, in case of partial necrosis, to target the areas of residual viable tumor. With spiral computed tomography or dynamic magnetic resonance imaging, residual viable hepatocellular carcinoma tissue is reliably depicted as it stands out in the arterial phase images against the unenhanced areas of coagulation necrosis. In the case of hypovascular metastases, a confident diagnosis of successful ablation can be made when an area of thermal necrosis exceeding that of the original lesion is depicted. Peripheral inflammatory reaction following ablation procedures should not be misinterpreted as tumor progression. PMID- 11379309 TI - Radiofrequency tissue ablation: physical principles and techniques for increasing coagulation necrosis. AB - Radiofrequency tumor ablation has been demonstrated as a reliable method for creating thermally-induced coagulation necrosis using either a percutaneous approach with image-guidance or direct surgical placement of thin electrodes into tissues to be treated. Early clinical trials with this technology have studied the treatment of hepatic, cerebral, and bony malignancies. The extent of coagulation necrosis induced with conventional monopolar radiofrequency electrodes is dependent on overall energy deposition, the duration of radiofrequency application, and radiofrequency electrode tip length and gauge. This article will discuss these technical considerations with the goal of defining optimal parameters for radiofrequency ablation. Strategies to further increase induced coagulation necrosis including: multiprobe and bipolar arrays, and internally-cooled radiofrequency electrodes, with or without pulsed radiofrequency or cluster technique will be presented. The development and laboratory results for many of these radiofrequency techniques and potential biophysical limitations to radiofrequency induced coagulation, such as perfusion mediated tissue cooling (vascular flow) will likewise be discussed. PMID- 11379310 TI - Resection of liver metastasis from gastric adenocarcinoma. AB - BACKGROUND/AIMS: To determine the factors influencing the prognosis of patients undergoing resection of liver metastases from gastric adenocarcinoma. METHODOLOGY: Over a 10-year period, at Kiryu Kousei General Hospital, 12 patients underwent potentially curative hepatectomy for metastatic adenocarcinoma of gastric origin. Two patients were excluded from this study, one because of postoperative death and one due to insufficient follow-up. We retrospectively examined the following factors: including TNM classification of the primary tumor, disease-free interval between gastric and hepatic resection, number and maximum diameter of the metastases, histological differentiation of the metastases, and the presence of lymphocyte aggregation enclosing the metastatic lesions. Survival rates were estimated by the Kaplan-Meier method and the weighting of each factor was compared by the log-rank test. RESULTS: The overall 5-year survival rate of the 10 patients was 10%. The median survival time after hepatectomy was 16.3 months, ranging from 3.1 to 245.7 months. Eight patients died of recurrent cancer and 1 died of unrelated septic shock with no evidence of cancer recurrence. Only one patient was alive without recurrence at the time of maximum follow-up. A significant survival advantage was noted in patients with disease-free interval > or = 1 year, and those with metastatic tumors < 5 cm in maximum diameter and/or enclosed by the aggregated lymphocytes, when compared with patients with disease-free interval < 1 year and those with metastatic tumors > or = 5 cm and/or directly infiltrated hepatic parenchyma. CONCLUSIONS: It was suggested that hepatectomy should be attempted in patients where the disease-free interval was > or = 1 year and with metastatic nodules < 5 cm. Lymphocyte aggregation around the metastatic tumor is a good prognostic sign for long-term survival. PMID- 11379311 TI - A proposal of no-touch isolation technique in pancreatoduodenectomy for periampullary carcinomas. AB - BACKGROUND/AIMS: The procedure of pancreatoduodenectomy for periampullary cancers accompanies a risk to shed cancer cells into a portal vein while handling the pancreas head lesion. This manipulation may subsequently cause a liver metastasis. We devised the no-touch isolation technique for pancreatoduodenectomy without removing a portal vein, for the purpose of preventing the manipulated shedding of cancer cells into a portal vein and liver metastasis. METHODOLOGY: The fundamental procedure of this technique is that isolation of portal vein precedes the handling of tumor mass. Isolation of a portal vein is carried out with the ligature of its surrounding veins after dividing of duodenum and pancreas. We applied the no-touch isolation technique for 10 cases, which consisted of 6 cases of distal bile duct carcinoma and 4 cases of ampullary carcinoma, between May 1993 and July 2000. RESULTS: There was neither operative mortality nor liver metastasis cases in these cases. CONCLUSIONS: The no-touch isolation technique without removing a portal vein might be recommended as a safe and reasonable procedure for periampullary cancer patients who have the potential for subsequent liver metastasis. PMID- 11379312 TI - Resection of liver metastasis after a pancreatoduodenectomy for pancreatic cancer: a case report. AB - A case of resected liver metastasis from pancreatic cancer, in a 69-year-old woman who obtained a relatively long survival, is presented. Primary pancreatic cancer was found in a patient presenting with obstructive jaundice and a loss of appetite. A pylorus-preserving pancreatoduodenectomy was carried out on December 2, 1996. Eleven months later, a 1.5-cm mass was found in the right posterior segment of the liver. Although the tumor size had increased to 2.5 cm 5 months later, no other metastases were observed. Therefore, the right posterior segment of the liver was resected. Eleven months after the second operation, recurrence was found in the cut surface of the liver and the left caudate lobe. She died on May 1, 1999. This case demonstrated a relatively favorable prognosis in comparison to other common type of liver metastasis from pancreatic cancer. This study discusses the peculiarities of this case and emphasizes the necessity to carefully select the patients before attempting to resect the liver metastasis from pancreatic cancer. PMID- 11379313 TI - Cytokines and acute-phase response markers derangements in patients with obstructive jaundice. AB - BACKGROUND/AIMS: Prolonged acute-phase response and increase of cytokines have been associated with higher mortality and surgical complications. This study investigated the status of cytokines and acute-phase response markers in patients with obstructive jaundice. METHODOLOGY: Forty-one patients were investigated. Endotoxin, tumor necrosis factor-alpha, interleukin-6, nitric oxide, C-reactive protein, liver enzymes, albumin and percentage of weight loss were determined at admission. RESULTS: Endotoxin, interleukin-6 and C-reactive protein were significantly elevated in both benign and malignant obstructive jaundice. Increased plasma levels of tumor necrosis factor-alpha were only detected in malignant tumors (68 vs. 24 pg/mL; P < 0.001). Patients with positive acute-phase response (C-reactive protein > mean + 2 SD of controls) had greater weight loss (P = 0.02), endotoxin (P = 0.03) and interleukin-6 plasma levels (P = 0.05) than those with no inflammatory response. Prolonged biliary obstruction (> 10 days) was associated with higher weight loss (P = 0.04), tumor necrosis factor-alpha (P = 0.003) and interleukin-6 (P = 0.05) plasma levels. CONCLUSIONS: A prolonged high-grade biliary tract obstruction prompted an increase in endotoxin levels, associated with a positive acute-phase response and cytokine elevation. PMID- 11379314 TI - Anomalous pancreaticobiliary ductal junction without bile duct dilatation in gallbladder cancer. AB - BACKGROUND/AIMS: Anomalous pancreaticobiliary junction is a rare anomaly but is a risk factor for primary carcinoma of the gallbladder. To define the relationship between anomalous pancreaticobiliary junction, especially if it is not associated with common bile duct dilatation, and gallbladder carcinoma, we retrospectively reviewed data of 126 patients with gallbladder carcinoma. METHODOLOGY: All these patients had undergone direct cholangiography either by endoscopic retrograde cholangiopancreaticography or percutaneous transhepatic cholangiography. RESULTS: Among 126 patients with gallbladder cancer, 23 patients (18.3%) exhibited anomalous pancreaticobiliary junction. Patients with anomalous pancreaticobiliary junction were younger (mean age: 54 +/- 9.1 years) than patients without anomalous pancreaticobiliary junction (mean age: 65 +/- 9.7 years). The incidence of gallstones in patients with anomalous pancreaticobiliary junction (17%) was significantly lower than in those without this anomaly (64%) (P < 0.01). Among the 23 patients with anomalous pancreaticobiliary junction, 12 patients (52%) had no bile duct dilatation and, 11 patients (48%) had bile duct dilatation in the form of fusiform or cylindrical dilatation. However, no cases with severe cystic dilatation were found. Patients of anomalous pancreaticobiliary junction without common bile duct dilatation had more advanced disease and poor prognosis than those with common bile duct dilatation. CONCLUSIONS: The present study revealed that gallbladder cancer in the patients of anomalous pancreaticobiliary junction without common bile duct dilatation was diagnosed at advanced stage and the prognosis was very poor. Therefore, if a minor abnormality is detected in the wall of acalculous gallbladder on ultrasonography, direct cholangiography should be done to exclude this anomaly. PMID- 11379315 TI - Endoscopic therapy in primary sclerosing cholangitis: outcome of treatment and risk of cancer. AB - BACKGROUND/AIMS: Primary sclerosing cholangitis is a cholestatic liver disease characterized by multifocal strictures in the intra- and extrahepatic biliary tree. Dominant strictures may arise in the extrahepatic bile ducts, and in these circumstances, endoscopic therapy has been introduced to relieve cholestasis and perhaps also delay the development of liver cirrhosis. The experience of endoscopic treatment at this point in time is limited and the long-term benefit is not clear. Neoplastic transformation in primary sclerosing cholangitis is unpredictable, which is illustrated in the present study along with an evaluation of the efficacy of endoscopic treatment. METHODOLOGY: Endoscopic retrograde cholangiopancreatography was performed in 25 patients with primary sclerosing cholangitis. In 15 there were dominant strictures in the hilum of the liver and/or the distal bile duct and these patients were treated by dilation and/or endoprostheses. Four patients in the treatment group had just cholestatic biochemical test results and 11 were symptomatic. RESULTS: Endoscopic therapy was technically successful in all 15 patients. In 43 sessions, 5 patients were treated by dilation, 2 with endoprostheses, and 8 by both methods. Improvement was achieved radiologically in 12 patients, clinically in 8, and according to liver function tests in 7. Therapy was complicated by cholangitis in 5 patients. Complications were mild and there was no mortality related to the procedure. However, 6 patients in the treatment group died, 5 of cholangiocarcinoma and 1 of colon cancer. CONCLUSIONS: Endoscopic therapy in primary sclerosing cholangitis is indicated in selected patients. The cancer incidence is high, not least in patients with deteriorating disease. It is important to find techniques for identifying patients at risk in order to perform liver transplantation before malignant transformation. PMID- 11379316 TI - Evidence of gallbladder function changes in hepatoma after transcatheter arterial embolization by quantitative Tc-99m DISIDA cholescintigraphy. AB - BACKGROUND/AIMS: Transcatheter arterial embolization is the treatment of choice for inoperable hepatocellular carcinoma. Gallbladder infarction following transcatheter arterial embolization has been reported, therefore, the gallbladder functions were studied using the quantitative Tc-99m DISIDA cholescintigraphy in the present study. METHODOLOGY: The gallbladder functions which were presented as the filling fraction and the ejection fraction in 24 patients with hepatoma before and after transcatheter arterial embolization. The patients were separated into two groups. Group 1: 12 patients received precystic artery transcatheter arterial embolization and group 2: 12 patients received postcystic artery transcatheter arterial embolization. RESULTS: After transcatheter arterial embolization, significantly decreased both gallbladder functions of filling fraction (61.2 +/- 7.4% and 48.3 +/- 6.5%) and ejection fraction (47.8 +/- 6.0% and 36.5 +/- 5.3%) were found in group 1 patients. However, no significant change of filling fraction (59.0 +/- 5.0% and 58.8 +/- 7.4%) and ejection fraction (49.9 +/- 2.4% and 49.3 +/- 5.7%) in group 2 patients. CONCLUSIONS: Impaired gallbladder functions were common in hepatoma patients who received precystic artery transcatheter arterial embolization, and Tc-99m DISIDA cholescintigraphy may be useful for evaluating the gallbladder functions in hepatoma patients who received transcatheter arterial embolization. PMID- 11379317 TI - Retroperitoneal multiple lymphangioma with differential cyst contents causing hydronephrosis and biliary dilatation. AB - We report a 40-year-old woman with retroperitoneal multiple cystic lymphangioma. Multiple cystic lesions were found on both sides of the aorta in the retroperitoneal and right pelvic spaces. These thin-walled cysts gradually increased in size leading to left hydronephrosis and dilatation of the common bile duct with cholecystitis, requiring resection. Analysis of cystic contents revealed differences between the cysts. The right cyst contained high concentrations of amylase, 3429 U/L, while the left cyst contained high concentrations of triglyceride, 470 mg/dL. No reports of patients with idiopathic retroperitoneal cystic lymphangioma have described differences in cystic contents. Differences in cystic contents according to tumor location strongly suggest that the cause of retroperitoneal lymphangioma is a developmental malformation in which lymphangiectasia follows the failure of developing lymphatic tissue to establish normal communication with the remainder of the lymphatic system. PMID- 11379318 TI - Obliterative arteritis with nitric oxide synthase and HLA-DR expression in Crohn's colitis. AB - BACKGROUND/AIMS: To cast light on whether inflammatory vascular injury is a possible pathogenic mechanism in Crohn's disease, the histological characteristics of vascular lesions were investigated. METHODOLOGY: Affected vessels in surgically resected colons from 23 patients with Crohn's disease, 20 with ulcerative colitis, 7 with ischemic colitis, and 9 normal controls were analyzed by Victoria blue and hematoxylin and eosin staining as well as immunohistochemistry for HLA-DR, nitric oxid synthase, vascular endothelial growth factor and E-cadherin. RESULTS: Inflammatory-cell infiltrates affecting arteries, accompanied by obliterative intimal thickening, were more frequent in Crohn's disease cases than in the other groups (P < 0.05-0.0001). Crohn's disease activity was positively correlated with the degree of obliterative arteritis. Granulomatous vasculitis was found exclusively in Crohn's disease (10 cases; 43.5%). In addition, focally enhanced endothelial staining of HLA-DR, with expression in granulomas adjacent to vessels was occasionally observed. In the endothelium of affected vessels, strong expression of HLA-DR was more prevalent in Crohn's disease and/or ulcerative colitis as compared with the ischemic colitis and controls (P < 0.05-0.01). In the involved arteries, enhanced endothelial nitric oxide synthase expression was most common in Crohn's disease among the groups (P < 0.05). A few cases of Crohn's disease, ulcerative colitis and ischemic colitis were positive for inducible nitric oxide synthase, vascular endothelial growth factor or E-cadherin in the vessel walls. CONCLUSIONS: The presence of characteristic obliterative arteritis and granulomatous vasculitis, a possible cause of ischemic injury, supports, in part, a vascular hypothesis for the pathogenesis of Crohn's disease. Enhanced expression of endothelial nitric oxide synthase and HLA-DR possibly reflects compensatory endothelium-mediated vasodilation and amplification of the immune response, respectively. PMID- 11379320 TI - IGF-1 stimulates mucosal growth in ileal Thiry-Vella fistulas. AB - BACKGROUND/AIMS: We determined whether the trophic effects of IGF-1 (Insulin-like growth factor-1) on the small bowel mucosa are mediated by either nonluminal factors or endogenous luminal secretion. The gut hormone IGF-1 stimulates growth of small bowel mucosa. The mechanisms responsible for this trophic effect are not known. METHODOLOGY: Rats underwent construction of a Thiry-Vella fistula of ileum. On postoperative day 10, the two groups were subdivided to receive either saline (control) or IGF-1. After 7 days, rats were killed and the Thiry-Vella fistula was removed. The mucosa was scraped and weighed, and protein content and alkaline phosphatase activity was determined. RESULTS: IGF-1 significantly increased mucosal weight and alkaline phosphatase activity and protein content of ileal Thiry-Vella fistula compared with the control rats. CONCLUSIONS: IGF-1 mediated stimulation of small bowel mucosal growth is mediated by factors that are independent of luminal contents and pancreaticobiliary secretion. IGF-1 may prove to be an important enterotrophic factor for gut mucosal proliferation. PMID- 11379319 TI - Flow cytometric DNA analysis, and immunohistochemical p53, PCNA and histopathologic study in primary achalasia: preliminary results. AB - BACKGROUND/AIMS: Primary achalasia is a premalignant disorder of the esophagus. The studies for esophageal cancer pathogenesis may reveal early diagnosis of esophageal cancer. DNA aneuploidy, p53 mutations and cellular proliferation are important factors in cancer development. As far as we know, we have not encountered any study on these factors in achalasia. METHODOLOGY: We studied DNA ploidy by flow cytometry and p53 and PCNA index by immunohistochemical technique and studied histopathology in the esophageal mucosa of primary achalasia and control patients. RESULTS: DNA analysis revealed aneuploidy in 2 of 20 achalasia patients but none of the 18 control patients. Sixty-five percent of achalasia and 22% of normal patients showed p53 positivity (P < 0.05). We have found normal mucosa, basal cell hyperplasia-esophagitis and dysplasia in 13, 22 and 3 patients and p53 positivity in 2, 12 and 3 of these patients, respectively (P < 0.05). PCNA labeling indexes (as % +/- SD) were 34.8 +/- 12.2, and 28.4 +/- 9.3 in achalasia and control groups, respectively (P > 0.05). PCNA labeling index was 28.0 +/- 8.2 in p53(-) and 36.0 +/- 12.9 in p53(+) patients (P < 0.05). PCNA indexes were found 29.3 +/- 9.6 in normal histopathologic group, 31.8 +/- 13.4 in basal cell hyperplasia-esophagitis, and 41.7 +/- 6.5 in dysplasia group (P > 0.05). CONCLUSIONS: DNA aneuploidy, p53 positivity, and higher cellular proliferation index may have important role in the pathogenesis of esophageal cancer in primary achalasia. PMID- 11379321 TI - Intestinal Behcet's disease associated with myelodysplastic syndrome with chromosomal trisomy 8--a report of two cases and a review of the literature. AB - Two cases of intestinal Behcet's disease, which developed in the state of myelodysplastic syndrome with trisomy 8, are presented. Both cases are included in the incomplete type of Behcet's disease, with recurrent aphthous stomatitis, skin lesions, genital ulcers or vascular involvement and punched-out ulcers in the cecum, without ocular involvement. The chromosomal analyses revealed chromosomal abnormalities, including trisomy 8, in both cases. Chromosomal trisomy 8 was shown in all 6 cases with the intestinal Behcet's disease associated with myelodysplastic syndrome reported previously, including our patients. Their histories indicated that myelodysplastic syndrome might have started before the development of intestinal Becet's disease. Theses findings suggested that chromosomal trisomy 8 might play an important role in the pathogenesis, at least in some groups, of intestinal Behcet's disease. PMID- 11379323 TI - Primary hepatic choriocarcinoma--a case report. AB - A 65-year-old man experienced anorexia and abdominal enlargement and died suddenly 45 days after admission. Multiple necrotic and hemorrhagic tumors were present in the liver (6 kg). The fragile tumor ruptured and a large amount of blood accumulated in the peritoneal cavity. The tumor was composed of cytotrophoblast and syncytiotrophoblast cells. Neoplastic cells were positive for human chorionic gonadotropin and negative for alpha-fetoprotein and carcinoembryonic antigen. The huge tumor directly invaded the entire antrum of stomach. Neoplastic trophoblastic cells proliferated mainly in the serosa and propria muscular layer of the stomach, and tiny cancer nests were observed in many vessels of the submucosa and propria mucosa. The serum alpha and beta human chorionic gonadotropin subunits showed a very high level; 51 ng/mL and 820 ng/mL, respectively. No tumor, cyst or scar was observed in the testes, mediastinum and retroperitoneum, macroscopically and microscopically. Based on these autopsy findings, we diagnosed this patient as having primary hepatic choriocarcinoma. PMID- 11379322 TI - Surgical treatment of pancreatic vasoactive intestinal polypeptide-secreting tumor: a case report. AB - A 68-year-old woman presented with secretory watery diarrhea causing hypokalimia, hypoalbuminemia and dehydration for 5 years. Subsequent investigations including abdominal ultrasonography and computed tomography scanning revealed a mass measuring 7 x 6 cm in the pancreatic tail. The diagnosis of pancreatic VIPomas was suspected on the basis of clinical symptoms. The patient underwent distal pancreatectomy and splenectomy after resuscitation of electrolyte imbalance, dehydration and malnutrition. The pathological examination with histoimmunochemical stain confirmed the diagnosis. Postoperative course is uneventful and the patient does not have symptoms any longer during the follow-up period. PMID- 11379324 TI - Combination therapy of percutaneous mitoxantrone injection, percutaneous ethanol injection, and transcatheter arterial embolization for intrahepatic hepatocellular carcinoma and adrenal metastasis. AB - We treated a 63-year-old man who had recurrent large hepatocellular carcinomas (> 5 cm in diameter) and left adrenal metastasis with the combination approach of percutaneous intratumoral chemotherapy with mitoxantrone, percutaneous ethanol injection, and transcatheter arterial embolization. He received repeated transcatheter arterial embolization and percutaneous ethanol injection combination therapy for intrahepatic hepatocellular carcinomas, which controlled his disease for 6 months from the first treatment. After that, left adrenal metastasis was detected by biopsy specimen. Therefore, we repeated more transcatheter arterial embolization and percutaneous ethanol injection to the liver and left adrenal gland, but this combination therapy could not control the hepatocellular carcinomas in these organs. With the patient's consent, he was treated with the combination approach of percutaneous intratumoral chemotherapy with mitoxantrone, percutaneous ethanol injection, and transcatheter arterial embolization for hepatocellular carcinomas of the liver and left adrenal gland. After this combination therapy, we followed-up the viable lesions by color Doppler ultrasonography and computed tomography examination. However, we could not detect these viable lesions of hepatocellular carcinomas in his body until one month before he died. When the degree of hepatic failure worsened due to the natural course of cirrhosis, this combination therapy was stopped 7 months before he died. He died of pulmonary tumor emboli from metastasis of inferior vena cava 24 months after the combination therapy started. However, on autopsy there was almost no remaining hepatocellular carcinoma found in the main lesions of liver and left adrenal gland. We suggest that a combination approach of percutaneous intratumoral chemotherapy with mitoxantrone, percutaneous ethanol injection, and transcatheter arterial embolization may be indicated in elderly cases of intrahepatic large hepatocellular carcinoma and adrenal metastasis, which are not under control only by transcatheter arterial embolization and percutaneous ethanol injection. PMID- 11379325 TI - Inferior vena cava injury after catheterization: report of a case. AB - We present a patient with retroperitoneal hematoma suspicious of inferior vena cava injury after catheterization for hemodialysis. Emergency computed tomography revealed extensive retroperitoneal hematoma and inferior vena cava angiography revealed extravasation. Emergent laparotomy was performed and repaired the perforation of inferior vena cava. His postoperative courses were uneventful and he remains well after the operation. PMID- 11379326 TI - Drainage for acute pancreatitis. AB - In severe acute pancreatitis, the drainage of activated pancreatic enzymes, infectious substances and necrotic tissue from the abdominal cavity is critical, especially after the operation. We use Faycon drainage tubes together with multi use feeding tubes. Just after the operation, normal saline is added through the feeding tube and the Faycon drainage tube is suctioned continuously. Irrigation is necessary for more than two weeks. PMID- 11379327 TI - The interval between flexible sigmoidoscopy screening examinations can be expanded beyond five years. AB - BACKGROUND/AIMS: As one of the methods for colorectal cancer screening, asymptomatic average-risk persons aged > or = 50 years are recommended to undergo flexible sigmoidoscopy screening every 5 years. We evaluate whether the interval between examinations can be extended beyond 5 years. METHODOLOGY: A total of 192 asymptomatic average-risk subjects were studied, all of whom had undergone a initial negative examination on a flexible sigmoidoscopy screening at age > or = 50 years and a second examination at least 3 years later. The study population was divided into three groups according to the interval between examinations, which was 3-5 years in Group A, 5-6 years in Group B, and 6-8 years in Group C. RESULTS: The incidence of neoplasms was compared among the three subjects groups, and it was found to be similar: 11/96 (11.5%) in group A, 4/55 (7.3%) in group B, and 5/41 (12.2%) in group C. All detected adenomas were less than 10 mm in diameter, and none contained a villous component or high-grade dysplasia. No cancers were found in the study. CONCLUSIONS: The results suggest that the interval for screening sigmoidoscopy may be extended beyond 5 years in persons showing negative results on an initial examination. PMID- 11379328 TI - Surgical outcomes for colorectal cancer patients including the elderly. AB - BACKGROUND/AIMS: The aim of this study was to compare the short- and long-term outcome of older and younger colorectal cancer patients resected for cure. METHODOLOGY: Three hundred and forty-six consecutive colorectal cancer patients who underwent some form of surgery were analyzed. One hundred and forty-four patients were < 65 years old (group 1), 151 patients were 65-79 years old (group 2), and 51 patients were 80 years or more (group 3). RESULTS: The overall perioperative mortality rate was 1.7% (n = 6). The median length of hospital stay was 19 days (range: 3-86 days). By univariate analysis, intraoperative bleeding (500 mL or more) (P = 0.009), duration of operations (240 min or more) (P = 0.03), and the presence of rectal cancer (P = 0.001), were strongly associated with higher incidence of postoperative complications. In multiple logistic regression analysis, only rectal cancer (P = 0.02) was significantly associated with serious postoperative complications. No age-related difference was noted concerning 5-year cancer-specific survival rates for patients with < 65, 65-79, and > or = 80 years who underwent surgery for cure (85%, 76%, and 69%, respectively) (P = 0.3). Using logistic regression analysis, tumor stage (P = 0.0001) and perioperative blood transfusions (500 mL or more) (P = 0.05) were strongly associated with outcome. CONCLUSIONS: Colorectal curative surgery for malignancy can be performed safely in the elderly with acceptable morbidity and mortality rates and long-term survival. PMID- 11379329 TI - Relationship between the fecal occult blood test and benign anal disorders. AB - BACKGROUND/AIMS: The fecal occult blood test is a very useful method for mass screening for colorectal cancer. The possibility of the fecal occult blood test being positive is high even in benign anal disorders, but the relationship between anal disorders and the fecal occult blood test has not been fully studied. METHODOLOGY: During the period from November 1995 to April 1996, we performed both fecal occult blood test and sigmoid colonofiberscopy for 440 patients with anal disorders who visited our hospital for the first time. RESULTS: The positive fecal occult blood test rate was 18.8% (77/409) for those who had only anal disorders and no findings in the colon and rectum. The positive fecal occult blood test rate was significantly high in the cases with polyps (42.3%) and in those with colorectal cancer (60%). CONCLUSIONS: These findings suggested the need to analyze fecal occult blood test, keeping in mind that fecal occult blood test might be positive even without any malignant findings in the large intestine in about 20% of cases of patients have anal disorders when fecal occult blood test is performed. PMID- 11379330 TI - The role of chemoembolization in the treatment of colorectal hepatic metastases. AB - In this brief review, we discuss the role of transcatheter arterial chemoembolization in the treatment and palliation of hepatic metastases from colorectal tumors. Although surgical resection is the optimal treatment for hepatic colorectal metastases, only a small percentage of these tumors are amenable to resection. A number of other modalities are being evaluated for these patients, of which, transcatheter arterial chemoembolization is the most promising. However, patients should be carefully selected and monitored to study outcomes; preferably these patients should be entered into randomized controlled trials. PMID- 11379331 TI - Endoscopic mucosal resection for early esophageal cancer and esophageal dysplasia. AB - BACKGROUND/AIMS: Advances in diagnostic technology have led to increased detection of early esophageal cancer, which is suitable for endoscopic treatment. We performed endoscopic esophageal mucosal resection of such cancer and dysplasia using the endoscopic esophageal mucosal resection tube and evaluated the clinical benefit of this technique. METHODOLOGY: Twenty-nine patients with esophageal mucosal cancer (27 cases with 33 lesions) or dysplasia (2 cases with 2 lesions) diagnosed between September 1992 and March 1998 were assessed endoscopically for the depth and extent of invasion by double staining with toluidine blue and iodine. Endoscopic ultrasonography was also performed to assess the depth of invasion in 22 cases with 22 lesions. RESULTS: The 35 esophageal lesions comprised 27 esophageal carcinomas and 8 areas of dysplasia. Twenty of the 35 lesions were resected en bloc and 15 were resected piecemeal. Subsequent surgery was performed for 5 cases with 7 lesions out of 10 cases with 15 lesions that were histopathologically diagnosed as m3 or more invasive. No recurrence has been detected in 24 evaluable cases (including 1 who died of another disease, 2 in whom surgery could not be performed due to complications, and 3 who refused subsequent surgery). No patients died of esophageal cancer after a mean follow-up period of 30.9 +/- 18.9 months. The 4-year survival rate was 100% in the m2 or less invasive group of 19 cases with 20 lesions, 75% in the m3 or higher invasive group of 5 cases with 8 lesions and 100% in the surgery group of 5 cases with 7 lesions (NS). No serious complications occurred except for 1 patient. Circumferential mucosal resection was done in this patient, resulting in esophageal stenosis, which responded to esophageal dilation. CONCLUSIONS: Esophageal mucosal resection using the endoscopic esophageal mucosal resection tube is safe and beneficial for early esophageal cancer and dysplasia. PMID- 11379332 TI - Pathogenesis of esophageal squamous cell carcinoma with lymphoid stroma. AB - BACKGROUND/AIMS: Lymphocyte infiltration in esophageal cancer, especially when beneath intraepithelial carcinoma, is frequently seen. However, cases of esophageal cancer with a dense stromal infiltration of lymphocytes are rare and the pathogenesis of such cases has yet to be clearly demonstrated. The objective of this study is to clarify its pathogenesis. METHODOLOGY: Four cases of esophageal squamous cell carcinoma with lymphoid stroma were investigated by immunohistochemical staining for the detection of Epstein-Barr virus, human papillomavirus, human leukocyte antigen-DR, as well as T and B cells in cancer tissue. RESULTS: In these four cases, neither positive staining of Epstein-Barr virus nor human papillomavirus infection was detected. On the other hand, the expression of human leukocyte antigen-DR antigen was evident in all cases with dense T-cell infiltration in the tumor tissue and moderate B-cell infiltration around the tumor. CONCLUSIONS: The expression of human leukocyte antigen-DR antigen without Epstein-Barr virus or human papillomavirus infection could thus be one possible pathogenesis of patients demonstrating esophageal squamous cell carcinoma with a lymphoid stroma. PMID- 11379333 TI - Prevalence and clinical presentation of subclinical/silent celiac disease in adults: an analysis on a 12-year observation. AB - BACKGROUND/AIMS: In recent years, an increased incidence of subclinical/silent celiac disease has been reported. Thus, we herein describe the prevalence and the clinical presentation of subclinical/silent celiac disease in 252 consecutive diagnosed celiac patients. METHODOLOGY: From 1988 to 1999 we diagnosed 252 celiac patients (74M and 178F, mean age: 27.9 yrs; range: 15-65 yrs, F/M ratio: 2.4). 144 patients were referred to us due to gastrointestinal symptoms, while 108 were referred to us from other specialists due to unexplained or unresponsive disease. All patients underwent both total immunoglobulin A and antigliadin antibodies antiendomysium antibody and evaluation, followed by gastrointestinal endoscopy with duodenal histological examination. RESULTS: 144 (57.14%) and 108 (42.86%) patients showed classical and subclinical/silent celiac disease, respectively. The most frequent extraintestinal marker of subclinical celiac disease were iron deficiency anemia (27.77%), alopecia and dermatitis herpetiformis (11.36%), osteoporosis (6.81%) and recurrent aphtous stomatitis (5.68%), while first-degree relatives (30%), Basedow's disease (25%) and insulin-dependent diabetes (20%) were the most frequent in silent celiac disease. CONCLUSIONS: This study confirms the extremely polymorphic nature of this condition that can affect several organs and apparatus without gastrointestinal symptoms. However, a more precise description of subclinical/silent celiac disease can only emerge from screening studies on general populations. PMID- 11379334 TI - Efficacy and tolerability of antibiotics in patients undergoing H. pylori eradication. AB - BACKGROUND/AIMS: Helicobacter pylori (H. pylori) infection is one of the most common gastrointestinal diseases. An increasing number of people undergo different treatment options. Unfortunately, H. pylori therapy may be troublesome for drug side effects and inefficacious for resistance to antibiotics. METHODOLOGY: One hundred and ninety-three (193) H. pylori-positive patients were randomly assigned to one of the following 7-day treatments: Group A (N = 64): amoxicillin, clarithromycin and rabeprazole; Group B (N = 64): tinidazole, clarithromycin and ranitidine bismuth citrate; Group C (N = 65): tinidazole, clarithromycin and rabeprazole Eradication was assessed by 13C-Urea Breath Test 6 8 weeks after the end of the therapy. Not-eradicated patients underwent a second cycle with tinidazole, tetracycline, bismuth and rabeprazole. All patients were asked to complete a validated questionnaire regarding presence and intensity of drug side effects. RESULTS: One hundred and eighty-eight out of the 193 H. pylori positive patients (96%) completed therapy. No significant difference in eradication rates was observed among the three groups both in intention to treat analysis and in per protocol analysis. No significant difference in incidence of side effects occurred among groups after the first-line regimens: 48% in group A, 44% in group B and 46% in group C. Twenty-two out of the 193 enrolled subjects (11%) were not eradicated after the first-line therapy. Among them, 86% were successfully eradicated by the tinidazole, tetracycline, bismuth and rabeprazole therapy. Moreover, during quadruple therapy, a higher prevalence and intensity of side effects than in each one of the groups submitted to the first-line triple therapy was observed. CONCLUSIONS: This study shows that triple rabeprazole-based eradicating regimens are effective and safe. Incidence of side effects seems low and similar in different three-drug regimens used. Quadruple therapy, which appear highly efficacious as a second line therapy, is associated with a significantly higher incidence of side effects when compared to first-line treatment. PMID- 11379335 TI - Polymorphism of tumor necrosis factor in esophageal, gastric or colorectal carcinoma. AB - BACKGROUND/AIMS: Several microsatellite polymorphisms located in the tumor necrosis factor locus on the chromosomal region 6p21.3 in the major histocompatibility complex region have been associated with malignant neoplasms and autoimmune diseases. In this study, we focused on the polymorphisms of tumor necrosis factor a and d from gastrointestinal carcinoma patients to ascertain whether they can be useful to predict these neoplasms. METHODOLOGY: We examined esophageal, gastric, and colorectal cancers (47, 53, 77 patients, respectively), and 213 normal controls. To compare the microsatellite polymorphisms of tumor necrosis factor a, d in Japanese individuals, dioxyribonucleic acids were extracted from normal mucosa (cancer patients) and from peripheral blood monocytes (the normal controls) by polymerase chain reaction. RESULTS: The frequency of tumor necrosis factor a3 allele was significantly higher in gastric cancer (P = 0.012) and that of tumor necrosis factor d7 allele was significantly higher in the colorectal cancer than the normal controls (P = 0.037). That of tumor necrosis factor a10 was significantly lower in the gastric cancer than the normal controls (P = 0.008). CONCLUSIONS: Microsatellite polymorphisms of tumor necrosis factor a and d might be significantly correlated with carcinogenesis of specific neoplasms, and may be useful for predicting these cancers. PMID- 11379336 TI - Extraintestinal infection by group C Salmonella: a case report and review of the literature. AB - Infected or mycotic aneurysms of the aorta are not very frequent but they are associated with high morbidity and mortality rates. Vascular infections due to Salmonella are not very frequent, but in recent years the reports of infections of this type have been on the increase. The authors report their experience with a case of aneurysm of the abdominal aorta infected by group C Salmonella and go on to review the literature on the subject. PMID- 11379337 TI - Intrahepatic portal vein branches after extrahepatic portal vein occlusion. Experimental study. AB - BACKGROUND/AIMS: Prehepatic portal hypertension caused by extrahepatic portal vein occlusion is a situation in which hepatocytes are not damaged by disease despite the fact that portal blood is unable to reach them due to portal vein occlusion. We explored the patency of intrahepatic portal vein branches after extrahepatic portal vein occlusion for the possibility of revascularization by splenoportal shunt. METHODOLOGY: Prehepatic portal hypertension was induced in 8 mini-pigs by external compression of the portal vein with a device consisting of an inflatable silicone balloon mounted on a silicone cuff and attached to a subcutaneous chamber. Another device consisting of cannula and a subcutaneous chamber was placed into the splenic vein for portal pressure monitoring and portal venograms. Both devices were placed during laparotomy with their chambers positioned subcutaneously. Portal vein compression was initiated one week later and was accomplished in two steps. Extrahepatic portal vein occlusion and the patency of intrahepatic portal vein branches were confirmed by direct portal venography. Alteration of the intrahepatic portal bed was examined at necropsy after 4 weeks, checking for the presence of occlusion or thrombosis. RESULTS: Portal vein occlusion was achieved in 5 animals, while severe stenosis was demonstrated in the remaining three. Portal venograms demonstrated patency of the lobar portal vein branches filled by hepatopetal collaterals around the occluded portal vein. All intrahepatic branches were free of thrombus at gross examination. CONCLUSIONS: In the absence of the hepatic parenchymal disease, lobar intrahepatic portal vein branches remain patent despite truncal portal vein occlusion and are supplied by rapidly developed hepatopetal collaterals. PMID- 11379338 TI - Risk factors for recurrence of large HCC in patients treated by combined TAE and PEI. AB - BACKGROUND/AIMS: In this report, risk factors of intrahepatic recurrence of a large solitary hepatocellular carcinoma after combination therapy with transcatheter arterial embolization followed by percutaneous ethanol injection were studied. METHODOLOGY: The series included 61 patients with an unresectable large solitary hepatocellular carcinoma, the largest size of which was greater than 3 cm in diameter. All patients completely responded to combination therapy and recurrence rates were determined. The following parameters; age, sex, hepatitis B virus surface antigen, hepatitis C virus antibodies, Child's classification, alcohol abuse, alanine aminotransferase, aspartate aminotransferase, alpha-fetoprotein, indocyanine green retention rate, hepatocellular carcinoma size, hepatocellular carcinoma capsule, total amount of injected ethanol and the alpha-fetoprotein 1 month after treatment were evaluated. RESULTS: The 1-, 3-, and 5-year cancer-free survival rates of all patients were calculated to be 61%, 23%, and 13%, respectively. Among pretreatment parameters, the log-rank test and subsequent Cox's proportional hazards model showed that a tumor size of more than 5 cm in diameter was independently associated with recurrence. The posttreatment parameters of total amount of injected ethanol was also shown to be significantly related to recurrence by the log-rank test. CONCLUSIONS: Lesions more than 5 cm in diameter and insufficient injected ethanol were associated with intrahepatic recurrence after this combination therapy. PMID- 11379339 TI - Liver transplantation with vena cava in situ and selective use of temporary portacaval shunt or portal clamping. AB - BACKGROUND/AIMS: The recipient hepatectomy with vena cava in situ in liver transplantation has overcome the need of venous-venous bypass thanks to temporary porta caval shunt or portal clamping. METHODOLOGY: 150 orthotopic liver transplants in 137 patients were performed and the vena cava in situ technique was used in 142 (venous bypass in 7, temporary porta caval shunt in 49, portal clamping in 87). The suprahepatic cava veins anastomosis was performed with Belghiti in 97 and piggyback techniques in 45. RESULTS: There were no differences in operative and warm ischemia times nor in blood requirements, while a greater stability of body temperature was documented in the vena cava In Situ group: in the latter temporary porta caval shunt preserved the temperature better than portal clamping (P < 0.01). In anhepatic phase mean artery pressure decreased in veno-venous bypass and increased in the vena cava In situ groups (P < 0.01). The venous return and the cardiac performances (anhepatic phase) were better preserved in the vena cava In Situ group. (P < 0.0001). CONCLUSIONS: Temporary portal caval shunt or portal clamping and piggyback or Belgiti Techniques allow a better hemodynamic stability through out the procedure, obviating the need for veno-venous bypass or fluid overload, if selectively used. PMID- 11379340 TI - Prophylactic chemolipiodolization for postoperative hepatoma patients. AB - BACKGROUND/AIMS: The efficacy of prophylactic chemolipiodolization following hepatic resection in patients with hepatocellular carcinoma was studied. METHODOLOGY: Forty-four of 67 consecutive patients with hepatocellular carcinoma who underwent hepatectomy between 1980 and 1997 were divided into two groups: group A (n = 21), in which prophylactic chemolipiodolization was performed during postoperative follow-up (2.4 times on average using a 39 mg mean dose of epirubicin or doxorubicin); and group B (n = 23), without prophylactic chemolipiodolization. The clinicopathological background and patient survival were compared retrospectively. RESULTS: There were no differences in the clinicopathological background between the two groups. Multiple intrahepatic recurrence was frequently observed in group B (P < 0.02). The recurrence-free survival rates in group A (54.4% and 31.1% at 3 and 5 years, respectively) were significantly higher than those in group B (15.7% and 7.9%, respectively). The survival rates of group A (95.2% and 80.4% at 3 and 5 years, respectively) were significantly higher than those in group B (40.1% and 22.9%, respectively). CONCLUSIONS: Our data suggest that postoperative prophylactic chemolipiodolization can be an effective treatment in reducing intrahepatic recurrence and may prolong survival for hepatocellular carcinoma patients following hepatic resection. PMID- 11379341 TI - Risk of developing hepatocellular carcinoma according to the titer of antibody to hepatitis C virus. AB - BACKGROUND/AIMS: Hepatitis C virus infection has been reported to be one of the main risk factor for developing hepatocellular carcinoma in Japan. The aim of the study was to examine the differences in the risk of developing hepatocellular carcinoma according to the titer of antibody to HCV. METHODOLOGY: A total of 13,173 inhabitants had their titers of anti-HCV examined based on a second generation passive hemagglutination assay, and we thus found 1,758 inhabitants whose anti-HCV titers were equal to or above 2(5). After carefully comparing our findings with the list of hepatocellular carcinoma in the Saga Prefectural Cancer Registry, we ascertained 45 cases of hepatocellular carcinoma (males 37, females 8). The logistic regression model was used to estimate the Odds ratio of risk factors for hepatocellular carcinoma. RESULTS: The risk of hepatocellular carcinoma in the subjects from 60-69 years of age was significantly higher than in the other age groups (Odds ratio = 3.88, P < 0.01). The risk of hepatocellular carcinoma in the males was also significantly higher than in the females (Odds ratio = 8.96, P < 0.001). The risk of hepatocellular carcinoma in the subjects with a titer of anti-HCV equal to or above 2(12) was significantly higher than in the subjects with a titer of less than 2(12) (Odds ratio = 33.46, P < 0.001). Furthermore, the age- and sex-adjusted risk for developing hepatocellular carcinoma for the subjects with a titer equal to or above 2(12) was significantly higher than that for subjects with a titer of less than 2(12) (Odds ratio = 32.56, P < 0.001). CONCLUSIONS: The risk of developing hepatocellular carcinoma was significantly high in the subjects with a titer equal to or above 2(12). To measure the titer of anti-HCV is thus considered to be useful for effectively detecting infection in a mass screening program. PMID- 11379342 TI - Procalcitonin is not an accurate marker of spontaneous bacterial peritonitis in patients with cirrhosis. AB - BACKGROUND/AIMS: The clinical features of peritonitis are usually absent in cirrhotic patients with an ascitic fluid infection, raising the interest for specific biological markers of inflammation. METHODOLOGY: We prospectively measured the plasma and ascitic fluid levels of procalcitonin, an innovative infection parameter, interleukin-6, and C-reactive protein in 20 cirrhotics with or without spontaneous bacterial peritonitis. The patient's condition was followed-up for 12 weeks after paracentesis. RESULTS: None of the 10 patients with spontaneous bacterial peritonitis presented with severe systemic signs of infection. Procalcitonin level in plasma, but not in ascites, was significantly higher in patients with spontaneous bacterial peritonitis compared to controls (0.74 +/- 0.6 vs. 0.2 +/- 0.1 ng/mL, P < 0.05). Interleukin-6 levels in ascites were similar between groups. C-reactive protein concentrations were higher both in plasma and in ascitic fluid in patients with spontaneous bacterial peritonitis compared to controls (85.3 +/- 63 vs. 18.6 +/- 19 mg/dL, 24.6 +/- 25 vs. 4.5 +/- 4 mg/dL, P < 0.05, respectively). Three patients with spontaneous bacterial peritonitis died, but the outcome was not related to the concentrations of biological markers. CONCLUSIONS: In spontaneous bacterial peritonitis, procalcitonin measurement is not an accurate diagnostic test, possibly due to the absence of systemic inflammatory response syndrome in this condition. In addition, the diagnostic value of C-reactive protein is limited by the wide overlap between values. PMID- 11379343 TI - Favorable quality of life after repeat hepatic resection for recurrent hepatocellular carcinoma. AB - BACKGROUND/AIMS: The appropriate choice of treatment for recurrent hepatocellular carcinoma after hepatic resection remains controversial. The aim of this study is to clarify prognostic factors and quality of life in patients with tumor recurrence after hepatic resection for hepatocellular carcinoma. METHODOLOGY: We retrospectively analyzed 188 patients with hepatocellular carcinoma who underwent curative hepatic resection between 1988 and 1997. Statistical analysis was performed to identify prognostic factors involved after recurrence. Furthermore, quality of life after treatment for recurrence was compared between patients with repeat hepatic resection or hepatic arterial infusion chemotherapy. RESULTS: In 123 patients with recurrence, unfavorable predictors after recurrence are pTNM Stage III/IV at initial surgery, receiving chemotherapy before initial surgery and presence of extrahepatic recurrence. In contrast, favorable predictors are 3 years or more of disease-free interval and repeat hepatic resection. The incidence of deteriorated performance status in the repeat hepatic resection group was lower than in the hepatic arterial infusion chemotherapy group because of better psychological function in patients undergoing repeat hepatic resection. CONCLUSIONS: Repeat hepatic resection provides a good prognosis and a favorable quality of life in patients with recurrence after hepatic resection for hepatocellular carcinoma. PMID- 11379344 TI - Improvement of hemorheological abnormalities in alcoholics by an oral antioxidant. AB - BACKGROUND/AIMS: It has been shown that alcohol impairs erythrocyte (red blood cell) membrane fluidity and lipid composition. The aim of this study was to test the effect of a novel acid-resistant antioxidant on the hemorrheology in alcoholics. METHODOLOGY: Thirty alcoholics (25 males, 5 females; mean age: 42 years; range: 31-54; 150 g ethanol/day for 3-5 years) were enrolled into the study. Patients were randomly and double-blindly allocated into 2 groups which were given, for a 2 week period, 18 g/day of Bionormalizer (obtained from biofermentation of carica papaya, pennisetum purpureum, sechium edule, Osato Res. Foundation, Gifu, Japan) dissolved in 5 mL of water at bedtime and 3 hours prior to examination. Placebo consisted of flavored sugar. Healthy teetotalers served as control. On the examination day, blood samples were taken for testing: routine tests, plasma glutathione, ascorbic acid, selenium, plasma lipid hydroperoxides and alpha-tocopherol. Erythrocytes were separated and tested for red blood cell malonyldialdehyde and glutathione content. The hemorheological studies were as follows: blood and plasma viscosity, whole blood filterability, red blood cell membrane fluidity by electron spin resonance, red blood cell aggregation index by photometric rheoscopy and red blood cell deformability by ektacytometry. RESULTS: As compared to healthy controls, alcoholics on placebo treatment showed no change of plasma viscosity but a significantly higher red blood cell malonyldialdehyde, blood viscosity (P < 0.05) and lower plasma glutathione, whole blood filterability and red blood cell fluidity (P < 0.01). No relationship appeared between biochemical tests and red blood cell membrane fluidity. Bionormalizer group showed a significant recovery to control values of either blood viscosity and whole blood filterability (P < 0.01) and a partial, although significant, improvement of red blood cell membrane fluidity, red blood cell malonyldialdehyde and plasma glutathione (P < 0.05). As compared to healthy control, red blood cell aggregation decreased in alcoholics (P < 0.05) and was not affected by Bionormalizer. However, Bionormalizer significantly improved the reduced red blood cell deformability (P < 0.05 vs. alcoholics) and this parameter correlated with red blood cell malonyldialdehyde (r: 0.62. P < 0.05). CONCLUSIONS: These preliminary data suggest that an effective antioxidant supplementation is able to improve the hemorrheology in alcoholics either by directly affecting the ethanol related lipoperoxidation and xanthine oxidase system activation and/or by modifying red blood cell membrane characteristics. PMID- 11379345 TI - Risk prediction using histology of noncancerous liver before hepatic resection for hepatocellular carcinoma. AB - BACKGROUND/AIMS: The aim of this study is to elucidate the feasibility of the risk assessment of hepatic resection by histological evaluation of noncancerous liver in patients with hepatocellular carcinoma. METHODOLOGY: The study involved 78 patients with hepatocellular carcinoma who had undergone a needle biopsy of noncancerous liver before hepatic resection. The histological activity index score which consists of four categories indicating the inflammatory activity and the degree of fibrosis was determined, and its association with complications after hepatic resection was examined. RESULTS: Postoperative complications occurred in 26 of the first 52 patients that underwent hepatic resection. A logistic analysis selected histological activity index score as an independent factor related to postoperative complications (Odds ratio 1.31, P < 0.02). Postoperative complications occurred more frequently in patients with a histological activity index score > or = 6 that had undergone resection of two or more segments (P < 0.05), and also in those with histological activity index score > or = 10 that had undergone segmentectomy or subsegmentectomy (P < 0.05). When the histological activity index score was taken into consideration in deciding operative procedures for a further 20 patients, the incidence of postoperative complications reduced considerably to 10%. CONCLUSIONS: Preoperative histological evaluation of noncancerous liver by a needle biopsy may be helpful in deciding the operative procedure to avoid complications after hepatic resection for hepatocellular carcinoma. PMID- 11379346 TI - Is hereditary hemochromatosis a balanced polymorphism: an analysis of family size among hemochromatosis heterozygotes. AB - BACKGROUND/AIMS: Hereditary hemochromatosis (HH) has a homozygote frequency of 0.5% and a heterozygote frequency of 13%, which suggests that hemochromatosis might be a balanced polymorphism. Evidence for this was sought by assessing reproductive success among hemochromatosis heterozygotes through determining their family size and incidence of miscarriage when compared to nongene carriers. METHODOLOGY: A cohort was defined in which the exposure variable was heterozygosity for hereditary hemochromatosis. Heterozygotes were identified by mailing individuals homozygous for hereditary hemochromatosis questionnaires concerning the number of siblings born alive and the number of miscarriages suffered by their mothers (predominantly heterozygotes, or HHH). Spouses of the hereditary hemochromatosis homozygotes were asked to complete accompanying questionnaires concerning their mothers (CONT). RESULTS: Five hundred and ninety six responses were received from hereditary hemochromatosis homozygotes and 532 responses from spouses of the hereditary hemochromatosis homozygote respondents. The mothers of the hereditary hemochromatosis homozygote respondents had a mean family size of 3.699. The mothers of the unexposed spouse controls a mean family size of 3.639 (P = 0.66 by ANOVA). One hundred and twenty-three of the HHH mothers suffered a total of 201 miscarriages, whereas 94 of the CONT mothers suffered 147 miscarriages (P = 0.38 by ANOVA). CONCLUSIONS: In this cohort, no evidence among hemochromatosis heterozygotes of reproductive advantage and therefore heterozygous advantage by this mechanism was seen. PMID- 11379347 TI - Serum leptin levels in patients with nonalcoholic chronic liver disease. AB - BACKGROUND/AIMS: The elevated serum leptin level of patients with alcoholic cirrhosis has been reported, however, the precise mechanism is still unknown. Leptin expression and protein synthesis have also been detected in activated hepatic stellate cells in cell cultures, which play a major role in hepatic fibrosis. We evaluated the serum leptin levels of patients with nonalcoholic liver diseases including cirrhosis and chronic hepatitis. We also investigated the hepatic clearance of leptin by determining the serum leptin level in blood samples obtained from the portal and hepatic veins. METHODOLOGY: The serum leptin level of 44 patients with nonalcoholic chronic liver disease (male/female = 21/23, cirrhosis/chronic hepatitis = 30/14) and 40 control subjects (male/female = 20/20) was determined in blood samples obtained from the antecubital vein by enzyme-linked immunosorbent assay. We also assessed the relationship between the leptin level and various biochemical tests of liver function. Additionally, we determined the leptin levels in the portal and the hepatic venous blood (nonalcoholic cirrhosis = 10, nonhepatic disease = 4). RESULTS: There were positive correlations between the serum leptin level and body mass index among males and among females in the liver disease group and in the control group. However, the serum leptin level of the liver disease group and control group did not differ significantly. Among the 44 liver disease patients, only the serum cholesterol level was significantly correlated with the serum leptin level after adjusting for sex and body mass index by multiple regression analysis. Furthermore, the leptin level in hepatic venous blood was significantly lower than that in portal venous blood. However, the ratio of [leptin level in hepatic venous blood]/[leptin level in portal venous blood] in the cirrhosis group, and that in the nonhepatic disease group, did not significantly differ. CONCLUSIONS: The serum leptin level of patients with nonalcoholic liver diseases is not elevated. On the other hand, the serum leptin level of patients with alcoholic cirrhosis has been reported to be elevated. The difference in the serum leptin level of patients with nonalcoholic liver disease and that of patients with alcoholic cirrhosis may be due to a difference in factors such as the levels of cytokines or sex steroids, and/or nutrition. Furthermore, it is likely that leptin is cleared in part by the portosystemic circulation through the liver. PMID- 11379348 TI - Effect of HSP70 induced by warm ischemia to the liver on liver function after partial hepatectomy. AB - BACKGROUND/AIMS: The purpose of this study was to determine if induction of HSP70 (heat shock protein 70), a stress protein which plays a cytoprotective role in response to various stimuli, protects hepatocytes from damage caused by partial hepatectomy and, if so, to elucidate the mechanism of such protection. METHODOLOGY: One hundred and eight male F344 rats weighing 190-220 g were randomly assigned to two groups with or without the presence of preconditioning. Fifteen-minute warm ischemia was applied to the liver of rats to induce HSP70, and 70% hepatectomy was performed 48 hours after the induction of HSP70 (ischemia group; n = 72). The rats in the nonischemia group did not undergo 15-min warm ischemia prior to 70% hepatectomy (nonischemia group; n = 36). Six rats, selected randomly from each group, were sacrificed at each measurement point to obtain blood and liver tissue samples. The levels of HSP70 in the liver, serum nitric oxide, levels of catalase and superoxide dismutase activity in the liver as antioxidative enzymes, and levels of Bcl-xL and Bax proteins and caspase-3-like activity in the liver as indices of apoptosis, were measured. RESULTS: The mean +/- SD level of HSP70 in the ischemia group (100 +/- 42 arbitrary unit (au)) was significantly higher than that of the nonischemia group (2 +/- 0.7 au) immediately before hepatectomy (P < 0.05). The ischemic preconditioning attenuated the liver damage caused by the subsequent partial hepatectomy. The levels of superoxide dismutase and catalase activity, serum nitric oxide level, and Bax protein level of the ischemia and nonischemia groups showed no significant differences after the partial hepatectomy. In contrast, the mean +/- SD level of Bcl-xL in the liver of the ischemia group (261 +/- 52 au) was significantly higher than that in the nonischemia group (114 +/- 33 au) 12 hours after the hepatectomy (P < 0.01). Furthermore, the mean +/- SD level of caspase-3 like activity in the liver of the ischemia group (18.1 +/- 4.6 au) was significantly lower than that of the nonischemia group (26.0 +/- 4.8 au) at 12 hours after the hepatectomy (P < 0.05). CONCLUSIONS: HSP70 induced by ischemic preconditioning prior to the partial hepatectomy was considered to protect the liver itself. In addition, the induced HSP70 may affect the Bcl-xL level after partial hepatectomy. Therefore, Bcl-xL seems to be involved in the reduction of liver damage after partial hepatectomy along with HSP. PMID- 11379349 TI - Helicobacter pylori infection does not correlate with plasma ammonia concentration and hepatic encephalopathy in patients with cirrhosis. AB - BACKGROUND/AIMS: In patients with cirrhosis, infection of the stomach with Helicobacter pylori may increase ammonia production and, consequently, the incidence of hepatic encephalopathy. To test this hypothesis a retrospective analysis was performed in patients with a transjugular intrahepatic portosystemic shunt. These patients are regarded to be ideal candidates for such a study since they have a high bioavailability of gut-derived ammonia and many of them develop spontaneous hepatic encephalopathy. METHODOLOGY: In 132 patients (Child-Pugh class A: 24%, B: 49%, C: 27%) with stable transjugular intrahepatic portosystemic shunt function for more than 3 months (mean follow-up: 15.5 +/- 10.8 months) the diagnosis of H. pylori infection was established by a specific and sensitive immunoblot assay for IgG- and IgA-antibodies. During follow-up, hepatic encephalopathy was assessed by clinical examination and a structured questionnaire. Venous plasma ammonia concentration was measured at the time of antibody determination (end of study period). RESULTS: Eighty-four patients (64%) had negative and 48 patients (36%) had positive immunoblots for H. pylori. The groups were comparable with respect to age, gender, etiology of cirrhosis, Child Pugh class, follow-up after transjugular intrahepatic portosystemic shunt, and shunt function. The ammonia concentrations of the patients without (group 1) and with antibodies against H. pylori (group 2) were 73 +/- 27 and 69 +/- 28 mumol/L (mean +/- SD), respectively. Hepatic encephalopathy occurred in 23 of 84 patients (27%) of group 1 and in 11 of 48 patients (23%) of group 2. CONCLUSIONS: A positive immunoblot for H. pylori antibodies neither correlates with plasma ammonia concentration nor with the incidence of hepatic encephalopathy in patients with cirrhosis of the liver and portosystemic shunt. PMID- 11379350 TI - Hepatocyte growth factor promotes liver regeneration and protein synthesis after hepatectomy in cirrhotic rats. AB - BACKGROUND/AIMS: Hepatocyte growth factor, a potent mitogen for hepatocytes has been reported to be a hepatrophic factor in normal livers. In this study, the effect of exogenous hepatocyte growth factor on liver regeneration in cirrhotic rats was investigated, in vitro and in vivo. METHODOLOGY: Liver cirrhosis was induced by intraperitoneal injections of an emulsion, carbon tetrachloride and olive oil, twice weekly for 10 weeks. In vitro, various amounts of exogenous hepatocyte growth factor; 0, 0.5, 1, 2.5, 5, and 10 ng/mL; were added to the hepatocytes isolated using in situ perfusion method. In vivo, partial hepatectomy (Hx), according to the procedure described by Higgins and Anderson, was performed on cirrhotic rats. Saline solution (control group) or 3 micrograms/kg of exogenous hepatocyte growth factor (HGF group) was then injected through the tail vein at intervals 12 hours after Hx. RESULTS: In vitro, DNA synthesis in hepatocytes obtained from cirrhotic livers increased following exogenous hepatocyte growth factor in dose-dependent fashion. In vivo, the labeling index of 5-bromo-2'-deoxyuridine at 24 hours after Hx was markedly increased by exogenous hepatocyte growth factor (control, 10.0 +/- 3.1%; hepatocyte growth factor, 25.8 +/- 9.8%; P < 0.01). Furthermore, serum albumin at 24 and 72 hours and a normotest at 24 hours after Hx, were significantly higher in the HGF group than in the control group. CONCLUSIONS: These results indicate that exogenous hepatocyte growth factor may promote DNA synthesis and protein synthesis during liver regeneration after Hx with cirrhosis. PMID- 11379351 TI - Asymptomatic portal vein obstruction after hepatobiliary resection: early detection by Doppler ultrasonography. AB - We report two different types of portal vein obstruction after liver resection: portal vein thrombosis due to steal phenomenon via a splenorenal shunt, and kinking of the skeletonized left portal vein after right hepatic lobectomy with caudate lobectomy. The two cases of portal vein obstruction were asymptomatic without any suggestive laboratory findings. Only routine Doppler ultrasonography detected portal vein obstruction which was successfully treated by emergency operation. PMID- 11379352 TI - Induction of T helper cell type 1 response and elimination of HBeAg during treatment with IL-12 in a patient with therapy-refractory chronic hepatitis B. AB - This study reports the increase of immunoregulatory T helper cell type 1 response and elimination of HBV-DNA during IL-12 therapy in a patient with chronic hepatitis B virus infection who had not responded to three previous interferon alpha therapies and one treatment with Famciclovir over a period of 6 years. The patient received IL-12 at a dose of 0.5 microgram/kg bodyweight weekly. Peripheral blood mononuclear cells were isolated before and during IL-12 application and incubated for 7 days with specific type 1 (purified protein derivative) and type 2 (tetanus-toxoid) TH cell antigens as well as with a macrophage/monocyte activating antigen (Bacille Calmette-Guerin). In the supernatants cytokines were determined by a double-sandwich ELISA. After 8 weeks HBV-DNA became negative and HBeAg seroconversion to anti-HBeAg occurred. Immunologically the loss of viremia was accompanied by a strong increase of the purified protein derivative-induced production of the type 1 cytokine interferon gamma (1219 pg/mL before, 13,138 pg/mL after IL-12 therapy). Furthermore, Bacille Calmette-Guerin-induced secretion of the macrophage/monocyte-associated cytokines IL-1, tumor necrosis factor-alpha and granulocyte-macrophage colony-stimulating factor was higher at the end of therapy. This case indicates that IL-12 enhances type 1 T helper cell activity which may be a predisposition for elimination of HBeAg and successful treatment of hepatitis B. PMID- 11379353 TI - Liver regeneration after hepatectomy. AB - Following hepatectomy, liver "knows" when to start and when to stop growing, and thereby accurately regulates its mass. Partial hepatectomy triggers hepatocyte proliferation whereas excessive liver mass (transplant) is regulated by apoptosis. Liver regeneration mainly involves the activation of adult hepatocytes and possibly of liver precursor ("stem") cells. The multistep process of liver regeneration comprises at least 2 critical phases: the transition of the quiescent hepatocyte into the cell cycle (priming) and the progression beyond the restriction point in the G1 phase of the cycle. The priming phase is characterized by the expression of immediate early genes. Activation of protooncogenes in the immediate early gene response involves both transcriptional and post-transcriptional mechanisms. Activation of four transcription factors, NF kappa B, STAT3, AP-1, and C/EBP beta, causes secondary activation of multiple genes and plays an important role in the initiation of hepatic regeneration. The passage of primed hepatocytes through the cell cycle is characterized by the expression of cell cycle genes and requires growth factors. An equilibrium between stimulator and inhibitor genes of the cell cycle expressed after hepatectomy, may explain why the liver regeneration is a tightly regulated growth process. Based on the knowledge of the regulation of liver regeneration, several practical considerations and potential therapeutic strategies can be applied in humans with hepatic dysfunction due to liver resection. PMID- 11379354 TI - Evaluation of gastroenteric bypass for unresectable pancreatic cancer. AB - BACKGROUND/AIMS: It is controversial as to whether gastroenteric bypass is helpful for patients with unresectable pancreatic cancer. This study was designed to evaluate the effects of gastroenteric bypass on dietary intake and the symptoms of gastric outlet obstruction in these patients. METHODOLOGY: We reviewed the cases of 101 patients with unresectable pancreatic cancer surgically treated at the Kobe University Hospital. The effects of gastroenteric bypass were examined by comparing the dietary intake and the symptoms of gastric outlet obstruction on admission, 1 month and 3 months after the operation. RESULTS: The analyses of dietary intake and the symptoms indicated that the gastroenteric bypass operation was not helpful for most of the patients with unresectable pancreatic cancer. Multivariate logistic regression model revealed that dietary intake on admission was the strongest parameter for dietary intake at one month after operation. The patients with a low dietary intake on admission often required a nasogastric tube after the bypass operation, reflecting progression of the disease. CONCLUSIONS: Gastroenteric bypass had no advantage to improve dietary intake and symptoms for almost all the patients with unresectable pancreatic cancer. It was effective only for patients with a high dietary intake without symptoms of gastric outlet obstruction on admission. PMID- 11379355 TI - Modified Devine exclusion for unresectable pancreatic head carcinoma. AB - BACKGROUND/AIMS: Gastrojejunostomy is generally performed for unresectable pancreatic head carcinoma. However, in the case of conventional gastrojejunostomy, the bypass does not always function effectively. METHODOLOGY: For unresectable pancreatic head carcinoma accompanied by severe duodenal stenosis, conventional gastrojejunostomy was performed in 5 cases, and modified Devine exclusion was performed in 7 cases. There were no significant differences between the groups regarding their backgrounds. RESULTS: There were no significant differences between the two groups for the average operation time, the days before peroral ingestion and the hospital stay. The state of peroral ingestion showed better results for modified Devine exclusion. The discharge rates were better for modified Devine exclusion, showing a significant difference (P = 0.028). The 50%-survival periods were 65 days and 159 days, respectively. The bleeding from the tumor occurred in 2 patients from the conventional gastrojejunostomy group, but none in modified Devine exclusion group. CONCLUSIONS: Modified Devine exclusion is a simple and effective technique for unresectable pancreatic head carcinoma. PMID- 11379356 TI - An apparent idiopathic case of relapsing acute pancreatitis. AB - We describe a case of relapsing acute pancreatitis apparently idiopathic in a 55 year-old man. The patient did not smoke and was a modest and irregular drinker of wine. Endoscopic retrograde cholangiopancreatography showed an initial dilatation of secondary ducts like a chronic pancreatitis of class I of Cremer. Ultrasound and computed tomography resulted negative for pancreatic lesions. In the follow up however, magnetic resonance cholangiopancreatography detected the presence of an intraductal mucin-hypersecreting neoplasm, a duct-ectatic mucinous cystic tumor of the pancreas, in the uncinate process. This is a benign lesion clearly recognized nowadays by magnetic resonance cholangiopancreatography, because this radiological technique shows the grape-like clusters of cystic lesions in secondary ducts communicating with the main duct on the same plane. The radiological picture above excludes a malignant lesion and a biopsy specimen is not required. Furthermore, an intraductal mucin-hypersecreting neoplasm of the pancreas does not require an immediate surgical resection because of its slow evolution and can be followed-up. Conversely cystoadenocarcinoma spreads in peripheral ducts and does not communicate with the Wirsung duct. It requires both surgical resection and a biopsy specimen for histological diagnosis. In the last episode of acute pancreatitis, a sphincterotomy was performed at endoscopic retrograde cholangiopancreatography and our patient had no more pain for one year. PMID- 11379357 TI - Two-staged treatment with local resection and percutaneous isolated hepatic chemoperfusion for advanced pancreatic cancer with multiple liver metastases: report of a case. AB - There have been disappointingly few effective treatment modalities for multiple liver metastases from pancreatic cancer. Percutaneous isolated hepatic perfusion, which was developed by us for delivering dose-intensive chemotherapy to the liver, has a high efficacy in the majority of patients with multiple primary and secondary liver tumors. We herein report the first experience of a two-stage treatment with extended local resection and subsequent two percutaneous isolated hepatic perfusions for advanced pancreatic ductal adenocarcinoma with liver metastases. The second percutaneous isolated hepatic perfusion with high-dose cisplatin and mitomycin G demonstrated a distinct regression of metastatic liver tumors. Although a long-term patient survival was not obtained due to local recurrence, liver metastases have been well controlled ever since. Given that further studies establish the efficacy of percutaneous isolated hepatic perfusion also in this field, this modality would be used as prophylaxis as well as treatment of liver metastasis in patients with advanced pancreatic ductal adenocarcinoma. PMID- 11379358 TI - Carcinoma of the head of the pancreas after excision of a choledochal cyst. AB - We here report a 53-year-old woman who had undergone resection of a choledochal cyst and hepaticojejunostomy three years before. She was readmitted because of intermittent fever, and abdominal computed tomography revealed a 4-cm tumor in the head of the pancreas. We performed pancreatoduodenectomy, and examination of the resected specimen showed well-differentiated papillary adenocarcinoma. Only 5 cases of carcinoma occurring after the resection of a choledochal cyst have been reported, and to our knowledge, this is the second case of carcinoma of the head of the pancreas. PMID- 11379359 TI - Langerhans cell histiocytosis of an adult with tumors in liver and spleen. AB - We describe a 58-year-old male with multiple histiocytic tumors in the liver and spleen. Multiple tumors in the liver and spleen were seen by image analysis, and splenectomy showed a large splenic tumor with a small nodule and a swelling lymph node in the hilus. Histological features of the tumors in the liver and spleen revealed proliferation of histiocytic cells with large and clear cytoplasm and a horseshoe-shaped nucleus. Immunohistochemical studies revealed the presence of S 100 protein and CD1a antigen in the tumor cells, and neither lymphocytic marker nor lysozyme was detected. No definite Birbeck granules were seen ultrastructurally, thus the tumor cells could be classified into Langerhans cell type without Birbeck granules. Administration of adriamycin, vincristine, cyclophosphamide and prednisolone reduced size and number of the liver tumors, and the histiocytic cells could not be detected in repeatedly biopsied tissue from liver tumor. We present the clinical, immunohistological and cytological features in a visceral type of adult Langerhans cell histiocytosis, which responded well to chemotherapy. PMID- 11379360 TI - Gastrectomized patients are in a state of chronic protein malnutrition analyses of 23 amino acids. AB - BACKGROUND/AIMS: Malnutrition is one of the major postoperative complications of radical subtotal or total gastrectomy for gastric cancer. This study was conducted to clarify the nutritional consequences of radical gastrectomy with respect to protein metabolism. METHODOLOGY: To evaluate the nutritional status and the abnormalities in protein metabolism in such cases, serum concentrations of 23 amino acids were measured by high performance liquid chromatography in 40 patients who had undergone either subtotal (n = 20) or total (n = 20) gastrectomy more than 6 months prior to this analysis. RESULTS: Serum concentrations of total amino acids and nonessential amino acids were the same between gastrectomized patients and healthy controls (n = 50). However, concentrations of essential amino acids, essential amino acid/nonessential amino acid and branched-chain amino acid/total amino acid ratios were significantly lower in patient groups than in normal controls. Each essential amino acid was decreased and concentrations of glutamate and citrulline were increased in both patient groups compared with controls. The major differences between patients with subtotal and total gastrectomies included an increased ornithine and a decreased arginine concentration in patients with subtotal gastrectomy. CONCLUSIONS: These changes suggest that malabsorption of protein from the intestinal tract causes persistent proteolysis in the skeletal muscle for long periods of time after surgery in these patients and that changes in ornithine and citrulline levels may reflect more severe alterations in those with total gastrectomy. PMID- 11379361 TI - Gastric myoelectrical activity in diabetics with and without diabetic autonomic neuropathy. AB - BACKGROUND/AIMS: The effects of diabetes mellitus on gastric myoelectrical activity has not been fully investigated. The aim of the present study was to investigate the pattern of gastric myoelectrical activity in noninsulin dependent diabetics, detected by electrogastrography in an attempt to clarify the relation between diabetic autonomic neuropathy and gastric myoelectrical abnormalities, if any. METHODOLOGY: The study was carried out on 34 noninsulin dependent diabetes (7 males, 27 females). Their age ranged from 35-60 years with mean age of 51.5 +/ 3.5 years. The EGG was recorded for 30 min in both the fasting and postprandial states, using an ambulatory EGG recording device (Digitrapper EGG, Synectics Medical). The adaptive spectral analysis method was used to assess the normality of the EGG. The EGG was defined as abnormal if: the percentage of normal slow waves (2.5-3.7 cycles/min) was below 70% during either the baseline or postprandial recording or there was a decrease in EGG peak power after the meal, or both. RESULTS: EGG abnormalities were detected in 13 patients (38.2%); 1 had tachygastria, 1 had bradygastria, 7 had dysrhythmias, and 4 had decreased EGG peak power after the meal. All diabetic patients with abnormal EGG suffer autonomic neuropathy. CONCLUSIONS: These results suggest that gastric myoelectrical abnormalities occur in a high proportion of noninsulin dependent diabetics and these abnormalities predominate in those patients with autonomic neuropathy. PMID- 11379362 TI - Is there any relationship between functional dyspepsia and chronic gastritis associated with Helicobacter pylori infection? AB - BACKGROUND/AIMS: The relationship between functional dyspepsia, H. pylori infection and chronic gastritis is controversial. Our aims were 1) To determine the prevalence of symptoms and the degree of association between symptoms and histopathological findings in different topographical gastric regions in patients with functional dyspepsia and H. pylori infection; 2) To determine the effect of eradication treatment on functional dyspepsia symptoms. METHODOLOGY: Prospective randomized study. 251 consecutive patients with dyspepsia (141 women and 110 men), mean age 48.08, SD 16.68 (without ulcer, gastric malignancy or reflux esophageal disease as determined by endoscopy), and with H. pylori infection, underwent upper endoscopy accompanied by the obtaining of 6 biopsies (cardia, corpus, antrum) at baseline, 3 and 6 months after treatment (pantoprazole 40 mg, once daily, amoxycillin 100 mg b.i.d., clarithromycine 500 mg b.i.d.). Inflammation, activity, H. pylori presence and other mucosal alterations were evaluated semi-quantitatively according to the Sydney system, before treatment and 6 months following treatment. An interview that was carried out before, and 6 months following the treatment, determined seven symptoms (scored as 0-3); epigastric burning and pressure, pain after meal, nausea, vomiting, bloating and belching, pain on empty stomach and anorexia. 95% confidence intervals were calculated for mean values of the symptoms and histological findings. The association between symptoms and histological findings was determined by the Kendall tau-b (K tau-b). Using the t test on a 5% level of significance we tested the null hypothesis that symptoms and histological findings were independent variables. RESULTS: The effectiveness of eradication after 3 months was 87.3% and after 6 months 92.0%. Reinfection rate after 6 months was 6.4% and the overall failure of eradication was 1.6%. Significant decline of chronic inflammation, activity and H. pylori was found in cardia, corpus and antrum (P = 0.001). Glandular atrophy was found to be lower in corpus and antrum (P = 0.001), whereas in cardia an increase was found. Intestinal metaplasia remained unchanged in all gastric regions, whereas a higher degree of foveolar hyperplasia was found, which was most pronounced in corpus and antrum (P = 0.01). There was a significant regression of lymphoid follicles in cardia and antrum (P = 0.001). On the first visit, the mean significant association between symptoms and histological findings was higher, with lower variation of K tau values as compared with the visit 6 months after treatment (K tau-b 0.171, SD 0.05, variation coefficient 30.5% vs. K tau-b 0.167, SD 0.07, variation coefficient 41.5%). According to the topographic distribution of gastritis at the time of the first visit, the mean significant association between symptoms and findings was found to be highest in antrum and corpus as opposed to the visit 6 months after treatment, where the values of association were found to be highest for variables from cardia and lowest for those in gastric corpus. After 6 months both the number of patients complaining of symptoms and dyspepsia score were lower (Wilcoxon P = 0.000). CONCLUSIONS: Advanced morphological changes of gastric mucosa were found to be significantly associated with symptoms of dysmotility. Pain on an empty stomach is predictive of antral inflammation. Cardia showed higher values of mean association with symptoms 6 months after therapy. Eradication treatment results in an improvement of both inflammatory changes and symptoms. In some patients persisting dysmotility symptoms were associated with persistent inflammation in cardia, which was also true for antrum, however to a lesser degree. PMID- 11379363 TI - Indication of splenectomy for gastric carcinoma involving the proximal part of the stomach. AB - BACKGROUND/AIMS: The role of splenectomy in the surgical management of gastric carcinoma is controversial and there is no consensus of opinion regarding the therapeutic value of splenectomy. The aim of this study was to search for possible metastasis to lymph nodes in the splenic hilum or along the splenic artery to avoid unnecessary splenectomy and to determine its indication. METHODOLOGY: The clinical records of 204 patients who underwent total gastrectomy combined with splenectomy for gastric carcinomas involving the proximal part of the stomach were analyzed. RESULTS: The incidence of nodal involvement to the splenic hilum and/or along the splenic artery was 49 (24.0%) of 204 gastric carcinomas involving the proximal part of the stomach that underwent combined gastrectomy and splenectomy. The characteristics of gastric carcinoma with metastasis to these nodes included a larger tumor, deeper penetration (T3, 4 tumors), a number of lymph node metastasis, and infiltrative type. In T2 cases, all the tumors with cancerous involvement to these nodes showed intraoperative gross serosal change). When the tumor size was less than 40 mm, nodal metastatic rate to the splenic hilum and/or along the splenic artery was very low. CONCLUSIONS: In conclusion, splenectomy should be conducted in T2 cases with gross serosal change and T3, 4 cases. With regard to tumor size, in the cases with a tumor whose size was less than 40 mm, it is possible to preserve the spleen in most cases. In the near future, splenectomy should be clarified precisely by randomized trials in advanced gastric carcinoma. PMID- 11379364 TI - Eosinophilic granuloma of the stomach mimicking gastric cancer, report of a case. AB - We report a rare case of eosinophilic granuloma of the stomach mimicking gastric cancer. A 49-year-old man was admitted to our hospital to undergo surgery for gastric tumor. Radiologic and endoscopic examination showed a protruding tumor with a deep ulcer at the anterior wall of the pylorus. Although malignant cells were not histologically confirmed in the biopsy specimens, subtotal gastrectomy with lymphadenectomy was performed because gastric cancer was strongly suspected. The gross appearance of the tumor seemed to be that of a gastric cancer, but the histological diagnosis was eosinophilic granuloma. If submucosal tumor of the stomach is suspected, eosinophilic granuloma should be considered as one of the differential diagnoses. Endoscopic removal of the tumor may be useful to make a precise diagnosis before surgery. PMID- 11379365 TI - Different patterns in the histology and autofluorescence of the Harderian glands of the Syrian Hamster, rat, mouse, Mongolian gerbil and guinea pig. AB - The purpose of this study was to compare the natural fluorescence in the Harderian glands of the Syrian hamster, rat, mouse, Mongolian gerbil and guinea pig (both sexes). For each species, 10 animals (five males and five females) were used. Histological autofluorescence studies were performed using a fluorescence microscope (450-490 nm filter). Two different types of fluorescent cells were observed in both hamster (type AFI high intensity and type AFII, low fluorescence) and rat (type AFI, low fluorescence and type AFII, high fluorescence) Harderian glands. The fluorescence was basally located in all mice cells, whereas it was observed near the epithelial cell nuclei in the Mongolian gerbil (occupying two-thirds and one-third of the cells in males and females, respectively). A high intensity of fluorescence was present throughout the acinar cells in the guinea pig. The patterns of fluorescence identified exhibited a sexual dimorphism in all species studied. These results demonstrate that the Harderian glands of the animal species examined exhibit a variety of histological autofluorescence patterns. PMID- 11379366 TI - Distribution of serotonin immunoreactivity in the main olfactory bulb of the Mongolian gerbil. AB - The distribution of serotonin immunoreactivity in the main olfactory bulb (MOB) of the Mongolian gerbil (Meriones unguiculatus) was examined by immunohistochemistry. Seven distinct layers of the Mongolian gerbil MOB-stained with cresyl violet were identified. Serotonin-immunoreactive (IR) cell bodies were not found in the MOB. The serotonin-IR nerve fibres had a specific laminar distribution and morphology in the gerbil MOB. Serotonin-IR nerve fibres were observed in the glomerular, external plexiform and granule cell layers of the MOB. These serotonin-IR nerve fibres showed varicosities that were larger than the thickness of the axon. The highest density of serotonin-IR nerve fibres was in glomeruli of the glomerular layer. The average fibre density in the glomerular layer was more than three to four times the density in the infraglomerular layers. Glomerular serotonin-IR fibres were much more intensively stained than infraglomerular serotonin-IR fibres. This result suggests that serotonin-IR nerve fibres of Mongolian gerbil MOB are extrinsic and may act to modulate the olfactory transmission. PMID- 11379367 TI - Distribution of lymphocyte subsets in the small intestine lymphoid tissue of 1 month-old lambs. AB - Distribution of lymphocyte subpopulations along the small intestine lymphoid tissue has been examined in 1-month-old lambs using flow cytometric and immunohistochemical techniques. Monoclonal antibodies against CD4, CD8, gamma delta, CD45R and B receptors have been employed in samples from continuous ileal Peyer's patch (IPP), discrete jejunal Peyer's patches (JPP), ileocaecal valve lymphoid tissue (ICVPP), mesenteric lymph node (MLN) and intra-epithelial (IEL) and lamina propria (LPL) lymphocytes. Histological studies were also done. Differences in the lymphocyte distribution have been observed between some of the regions examined, especially between IPP and JPP for most of the markers. A remarkable feature was the existence of morphological and lymphocyte distribution differences between ICVPP and IPP, locations that had been traditionally considered as similar. The antibody against CD45R receptor used in this study, that was supposed to mark B cells and some T cells, detected cell populations located in the dome of the follicles in all the samples, whereas the centre was negative. Lymphocytes positive to the B marker employed were located mainly in the centre, suggesting that both antibodies would mark B cells in different maturation status. PMID- 11379368 TI - [The functional morphology of the large intestinal mucosa of the ox (Bos primigenius f. taurus), sheep (Ovis ammon f. aries) and goat (Capra aegagrus f. hircus)]. AB - The mucous membrane of the large intestine of the ox, sheep and goat was studied using light and electron microscopy. The surface epithelium possesses the well known complement of organelles with a conspicuous number of mitochondria as a source of energy for absorptive activities. Occasionally, brush cells are found in the epithelium; the functional significance of this type of cell is still under discussion. In the present material, electron-lucid spheroids (diameter: 45 x 35 nm), which tend to fuse, appear in the supranuclearly located osmiophilic granules. The structure and appearance of the epithelium cells in the crypts of these species correspond to previous descriptions in other mammals. In the epithelial cells of the deep glandular region, dense accumulations of mainly rounded granules (diameter up to 400 nm) are found apically in the cytoplasm. Their functional significance is still under discussion. Acid phosphatase activity, as a marker enzyme for lysosomes, points to degenerative processes in connection with the physiological death of enterocytes. In contrast, the negative alkaline phosphatase reaction indicates that, according to our findings, the epithelial transport of substances in the large intestine of ruminants--in principle contrary to the situation in the small intestinal enterocytes--occurs without the aid of this enzyme. The demonstrated ATPase activity on the other hand, is related to a primary active ion transport linked with membrane exchange of non-electrolytes in connection with transepithelial events of absorption and secretion in the large intestine. PMID- 11379369 TI - Light and electron microscope studies on the nasopharynx and nasopharyngeal tonsil of the horse. AB - Light and electron microscope studies were conducted on the nasopharynx and the nasopharyngeal tonsil of 15 young horses. The nasopharynx and nasopharyngeal tonsil was lined with pseudostratified columnar ciliated epithelium and goblet cells. The lymphoepithelium of the nasopharyngeal tonsil was folded forming crypts, the mucosa of which was modified into follicle associated epithelium characterized by stratified cuboidal epithelium, loss of cilia, absence of goblet cells and infiltration of lymphocytes. The lamina propria mucosae of the nasopharyngeal tonsil contained well-developed lymphoid tissue and clusters of seromucus acini. Scanning electron-microscopy revealed a dense mat of cilia covering the nasopharynx and nasopharyngeal tonsil. The follicle-associated epithelium consisted of different populations of microvillus cells in addition to M cells with very short microvilli and a few squamous and intermediate cells. Microvillus cells in the deeper part of the FAE had larger microvilli and their cytoplasm contained a dense population of mitochondria, smooth and rough endoplasmic reticulum, Golgi complexes and lysosomes. The flat surfaced M cell had a more electron-dense cytoplasm and contained small supranuclear vacuoles in addition to the organelles seen in microvillus cells. PMID- 11379370 TI - The intermesenteric plexus in the pigeon (Columba livia GM). AB - Using the thiocholine method and histological techniques the intermesenteric plexus of the pigeon was studied. The intermesenteric plexus of this species is a plexo-ganglionic structure with several (five to seven) ganglia and nerve fibres. The ganglia have an oval-, spindle- or star-like shape. Single nerve cells along the nerve fibres were observed. The intermesenteric plexus of the pigeon is situated on the ventrolateral surface of the aorta, between the cranial mesenteric artery and caudal mesenteric artery. The connections between the intermesenteric and other vegetative plexuses (coeliac plexus, mesenteric inferior plexus) and the pelvic nerve were observed. PMID- 11379371 TI - [Principals of construction in the forepaw and hindpaw of the domestic cat (Felis catus). III: Musculature]. AB - With regard to muscular mass and inner structure, the muscles of the forepaw and hindpaw of the domestic cat are grouped functionally, compared and evaluated. A high degree of functional adaptation is present in these muscles, for example in the passive locomotion apparatus of the paws. The domestic cat is not a 'running animal' like other domestic mammals. Also, in domestication it has preserved a strongly individual way of life and has remained a quick, ambushing hunter and climber on the run. Hence its forepaw is a flexible guiding and gripping apparatus, while its hindpaw provides stability for sprinting and non-skid pushing. The partly re-interpreted functions of the muscles are described and listed in four tables. PMID- 11379372 TI - The hot flush: the enigma of the climacteric. PMID- 11379373 TI - Effects on menopausal symptoms and acceptability of isoflavone-containing soy powder dietary supplementation. AB - OBJECTIVES: To assess the acceptability of the delivery of an isoflavone supplementation in the form of a powdered drink, and whether the supplementation of dietary isoflavones in this manner decreased the incidence of menopausal flushes. The secondary aims included assessment of other symptoms or parameters of estrogen deficiency and responses to isoflavones. METHODS: A randomized, double-blind, placebo-controlled, parallel-group trial comprising 24 postmenopausal women with symptoms of estrogen deficiency was performed over a 12 week period. The women were randomized to receive a dietary beverage containing isoflavones or an isoflavone-free, isocaloric placebo preparation. RESULTS: Although there was a high compliance rate among individual patients, there was a 25% withdrawal rate from the study in the active group. The incidence of complaints of bad taste tended to be higher in the active group (p = 0.07), and the total number of adverse events was significantly higher in this group (p < 0.001). There was no statistically significant difference in the incidence of flushes between the groups. There was no difference between the groups in Greene Menopause Symptom Scores, vaginal maturation value, levels of follicle stimulating hormone (FSH) or sex hormone-binding globulin (SHBG), or bone turnover markers. CONCLUSIONS: Powdered energy drinks are not commonly consumed in Australia and were poorly tolerated in this study. The high withdrawal rate and reporting of side-effects suggests that other methods of isoflavone delivery may be more appropriate in this culture, in future trials. At the dose used no benefit was seen in relief from menopausal symptoms, although for the sample size, the study could only have been expected to detect major differences between the groups. PMID- 11379374 TI - Perimenopausal women in estrogen vasomotor trials: contribution to placebo effect and efficacy outcome. AB - OBJECTIVE: To determine the difference, if any, in the placebo response when both perimenopausal and postmenopausal women are enrolled, compared with postmenopausal women alone, in a study assessing the efficacy of synthetic conjugated estrogen tablets on moderate-to-severe vasomotor symptoms (MSVS). METHODS: A total of 120 healthy women (72 active; 48 placebo) complaining of moderate-to-severe vasomotor symptoms were enrolled in a randomized, placebo controlled, double-blind, multicenter clinical trial. In all, 109 patients completed treatment to week 12. Women were enrolled using minimal inclusion and exclusion criteria, and included perimenopausal women (n = 34; 0 to 6 months since last menses) as well as menopausal women (n = 79; > 12 months since last menses). RESULTS: Changes in MSVS in the intent-to-treat (ITT) population showed differences between the active and placebo treatments at weeks 4 (p < 0.022), 8 (p < 0.010) and 12 (p < 0.010). At week 12, the mean percentage reduction in MSVS was 81% for the active treatment group and 58% in the placebo treatment group. To examine the placebo response, the ITT population was broken down into groups defined by the time since their last menses. The perimenopausal group (0 to 6 months since last menses) demonstrated a consistently higher placebo response than that of the postmenopausal groups (> 12 months since last menses). CONCLUSIONS: Perimenopausal women contributed to a higher placebo response, compared with the rate of response previously reported in clinical studies of estrogen replacement in postmenopausal women. Including perimenopausal women in future vasomotor symptom trials will require study populations of sufficient size to maintain the statistical power to demonstrate a difference between therapeutic response to active or placebo treatment. PMID- 11379375 TI - The effects of tibolone on vaginal blood flow, sexual desire and arousability in postmenopausal women. AB - AIMS: To compare the effects of 3 months' tibolone treatment with the effects of placebo on sexual function (in particular, vaginal blood flow, and sexual desire and arousability) and climacteric symptoms in postmenopausal women. METHODS: A randomized, double-blind, cross-over study was conducted in 38 postmenopausal women who received tibolone 2.5 mg/day and placebo. Vaginal blood flow during erotic stimulation by fantasy and film was measured using a vaginal photoplethysmograph and subjects completed sexual function questionnaires and daily diaries. RESULTS: Tibolone significantly increased baseline vaginal pulse amplitude (VPA) levels compared with placebo. There were significant treatment differences in VPA in favor of tibolone during fantasy periods but not during erotic film stimulation. Tibolone was associated with significant increases in sexual desire, and the frequency of arousability and of sexual fantasies compared with those with placebo. Vaginal lubrication was significantly improved on tibolone. Twenty-five of 38 (66%) subjects correctly guessed when they were on active treatment. Tibolone was well tolerated. CONCLUSIONS: Tibolone was associated with significant improvements in sexual function in postmenopausal women, reflecting both its estrogenic and androgenic properties. There were significantly greater increases in vaginal blood flow with tibolone in response to erotic fantasy but not film, suggesting two possible pathways of female sexual response. PMID- 11379376 TI - Female sexual function and dysfunction: can the Cinderella of medicine turn into a princess? PMID- 11379377 TI - Mammographic breast density during hormone replacement therapy: effects of continuous combination, unopposed transdermal and low-potency estrogen regimens. AB - OBJECTIVE: The aim of this study was to evaluate the impact of different hormone replacement therapy (HRT) regimens on mammographic breast density. STUDY DESIGN: Mammographic density was recorded in women participating in a population-based screening program. At first mammogram, all women were non-users of HRT, and thereafter reported continuous use of the same HRT regimen. The study population comprised 158 women: a total of 52 women were using continuous combined HRT (conjugated equine estrogen 0.625 mg plus medroxyprogesterone acetate 5 mg); 51 women were using low-dose oral estrogen alone (estriol 2 mg daily); and 55 women were using unopposed transdermal estrogen given as a patch (estradiol 50 micrograms/24 h). Films were coded and analyzed for mammographic density by an independent radiologist blinded to treatments. Mammographic density was classified according to Wolfe. RESULTS: An increase in mammographic density was much more common among women taking continuous combined HRT (40%) than for those using oral low-dose estrogen (6%) and transdermal (2%) treatment. The increase in density was already apparent at the first visit after starting HRT. During long term follow-up, there was very little change in mammographic status. CONCLUSION: HRT regimens were shown to have different effects on the normal breast. There is an urgent need to clarify the biological nature and significance of a change in mammographic density during treatment and, in particular, its relation to symptoms and breast cancer risk. PMID- 11379378 TI - Antiatherogenic effects of 17 beta-estradiol and 17 alpha-estradiol and its derivative J811 in cholesterol-fed rabbits with thyroid inhibition. AB - OBJECTIVE: The aim of this study was to investigate the antiatherogenic effects of 17 beta-estradiol and 17 alpha-estradiol and its derivative J811 (estra 1,3,5(10),8-tetraene-3, 17 alpha-diol), having a non-feminizing effect and high antioxidant potential, in male rabbits. EXPERIMENTAL DESIGN: Male White-Russian rabbits weighing 2.1-2.6 kg were fed either a standard or a high-cholesterol (200 mg/kg) diet, with thyroid function-inhibiting thiouracil (20 mg/kg) combined with cholic acid (40 mg/kg) administered daily in sunflower oil for 3 months. During the last month of the study, estrogens were administered by gavage at a dose of 0.02 or 0.1 mg/kg. RESULTS AND CONCLUSIONS: All three estrogens exerted remarkable antiatherosclerotic effects. Decreases in serum and aortic-wall lipid parameters and the index of atherogenicity were dependent on estrogen dose. Morphological evaluation of the aortic wall (height of plaques, size of plaque relative to aortic half-circumference) showed only weak therapeutic effects with all three estrogens. It is an open question whether the treatment period was too short to reverse the above changes. On the other hand, the data clearly suggest that 17 alpha-estradiol and J811 offer new perspectives for the prevention of atherosclerosis in men, which is similar to that found with 17 beta-estradiol in women. PMID- 11379379 TI - Oral estrogen replacement therapy versus placebo for hot flushes: a systematic review. AB - OBJECTIVE: To examine the effectiveness of oral hormone replacement therapy (HRT) with regard to hot flushes and night sweats, which are vasomotor symptoms commonly experienced by menopausal women. METHODS: Double-blind, randomized, placebo-controlled trials of oral HRT for at least 3 months' duration and reporting vasomotor outcomes were identified using a search strategy developed by the Menstrual Disorders Group and Subfertility Group of the Cochrane Collaboration. Twenty-one trials meeting the selection criteria were included in the review. Study participants totalled 2511. Trial duration ranged from 3 months to 3 years. RESULTS: There was a significant reduction in the weekly hot flush frequency for HRT compared with placebo (weighted mean difference -17.46, 95% confidence interval (CI) -24.72 to -10.21). This was equivalent to a 77% reduction in frequency (95% CI 58.2-87.5) for HRT relative to placebo. Symptom severity was also significantly reduced compared with placebo (odds ratio (OR) 0.13, 95% CI 0.08-0.22). Withdrawal for lack of efficacy occurred significantly more often with placebo therapy (OR 17.25, 95% CI 8.23-36.15). Withdrawal for adverse events, commonly breast tenderness, edema, joint pain and psychological symptoms, was not significantly increased for HRT (OR 1.38, 95% CI 0.87-2.21). In women who were randomized to placebo treatment, a 50.8% (95% CI 41.7-58.5) reduction in hot flushes was observed between baseline and the end of the study. CONCLUSIONS: Oral HRT is highly effective in alleviating hot flushes and night sweats. Owing to the marked effect seen with placebo treatment, it is important to compare therapies purported to alleviate vasomotor symptoms with a placebo or an established therapy. PMID- 11379380 TI - Breast epithelial proliferation in postmenopausal women evaluated through fine needle-aspiration cytology. AB - OBJECTIVE: The aim was to evaluate fine-needle-aspiration (FNA) cytology as a method of following breast epithelial proliferation in postmenopausal women during hormone replacement therapy (HRT). METHODS: Twelve healthy postmenopausal women were recruited and randomized to two different types of sequential HRT during 4 months of treatment. The women were administered continuous estradiol 50 micrograms/24 h with the addition of progestogen sequentially in the form of either vaginal progesterone gel 8 mg every 2nd day or medroxyprogesterone acetate 5 mg/day orally during 12 days per month for the complete treatment period. Fine needle-aspiration biopsies were performed twice during the estrogen phases and twice during the estrogen plus progestogen phases of treatment. Breast epithelial proliferation was analyzed in these samples by immunocytochemistry to measure the content of the nuclear antigen Ki-67/MIB-1, which is expressed in proliferating cells. RESULTS: From the 12 women, a total number of 47 FNA biopsies were taken. Thirty-eight of these aspirates, 19 from each of the estrogen and the estrogen plus progestogen phases, were evaluable for MIB-1 content (81%). There was a non significant increase in levels of proliferation during the combined estrogen progestogen phase (2.1%) compared with the estrogen-only phase (1.4%). These values were similar to those previously observed during the menstrual cycle in young fertile women. CONCLUSIONS: We conclude that the FNA biopsy technique is feasible for studying proliferation not only in young, normally cycling women but also in the postmenopausal breast. PMID- 11379381 TI - Atrophic vaginitis in Filipino women. PMID- 11379382 TI - The rationale for a wider range of progestogens. AB - Progestogens are commonly used in hormone replacement therapy, normally as opponents of estrogen to protect the endometrium from hyperplasia and cancer. While these benefits of endometrial protection are well recognized, the data related to the effect of progestogens on breast tissue and the cardiovascular system are conflicting. It has been demonstrated that, according to the type of progestogen used, and the dose and duration of its application, a predominant proliferative effect may be observed in human breast cells. As far as breast cancer is concerned, most epidemiological studies suggest no difference in risk between therapy with estrogens alone or estrogens combined with progestogens, but recent data do indicate an increased risk with combined therapy. When the cardiovascular risk factors are considered, some progestogen molecules with a higher androgenic potency than others attenuate the beneficial effects of estrogens on both the lipid profile and vasomotion. On the other hand, the epidemiological data on primary prevention do not suggest any negative effect of the progestogens administered together with estrogens on cardiovascular morbidity or mortality. Recent results have questioned the cardioprotective effect of hormone replacement therapy in women with established coronary heart disease. It has been suggested that the lack of a secondary preventive effect by hormone replacement therapy may be due to the progestogens selected. The effect on osteoporosis is also the subject of debate, with some progestogens having a neutral effect on bone mineral density and others producing a marked improvement. Awareness of the classic contraindications of hormone replacement therapy and selection of molecules devoid of estrogenic, androgenic or glucocorticoid effect should allow greater use of the progestogens without any major drawback. PMID- 11379383 TI - The role of antiandrogens in hormone replacement therapy. AB - The most widely used antiandrogens in clinical practice are cyproterone acetate, a derivative of 17-hydroxyprogesterone, and dienogest, a 19-norprogestin. An established sequential preparation for hormone replacement therapy (HRT) consists of 11 days of 2 mg estradiol valerate, 10 days of 2 mg estradiol valerate with 1 mg cyproterone acetate, followed by a 7-day drug-free interval (Climen, Schering AG, Berlin, Germany). Cyproterone acetate is highly progestogenic, has no androgenic properties, and is antiandrogenic above a certain dose. Cyproterone acetate does not counteract the estrogenic effects of estradiol valerate in Climen. This therapy, therefore, has optimal effects on lipid metabolism and coronary heart disease risk, protects the endometrium and reduces menopausal symptoms, preserves bone and reduces osteoporotic fracture risk, and has antiandrogenic effects on the skin and other androgen-sensitive epidermal structures. Dienogest, on the other hand, will soon be introduced in a continuous combined HRT. Dienogest has a 17 alpha-cyanomethyl group instead of the 17 alpha ethinyl group typical of the common 19-nortestosterone derivatives. It is also referred to as a hybrid progestogen because it has pharmacodynamic properties (e.g. antiandrogenicity) in common with progesterone derivatives. A fixed formulation containing 2 mg estradiol valerate and 2 mg dienogest (Climodien) for continuous combined HRT has been developed. This formulation had excellent effects on vasomotor and neurovegetative symptoms. The bleeding pattern was generally highly satisfactory and similar to that with Kliogest, as were the results of endometrial biopsies after 12 cycles of treatment. Lipid metabolic changes may be interpreted as beneficial. Dienogest had no adverse effects on the vasorelaxant effect of estradiol valerate in postmenopausal women, as shown by markers of vascular function. Neuropsychological studies utilizing evoked potentials showed shortening effects on sleep latency and an improvement in cognitive information processing. Continuous combined HRT with dienogest, therefore, may come to be regarded as the HRT of choice in postmenopausal patients with mood defects. In summary, HRT with antiandrogenic progestogens has its specific indications with respect to preserving metabolic estrogenicity, specific antiandrogenic effects and specific effects on vigilance and mood disorders. PMID- 11379384 TI - The need for continuing education of the prescriber. AB - The Millennium will be seen as an important time of change in the doctor-patient relationship. Until very recently, many patients, male and female, expected not only advice from their doctor, but also to be told which treatment would be best for them. This paternalistic approach, however, is rapidly disappearing. Nowadays, early post-menopausal women expect to make decisions about their treatment, after being fully informed of all the possible benefits and risks. Provision for this requires the doctor to keep abreast of the literature. Occasionally, new data are released that challenge established beliefs. It was thought, for example, that hormone replacement therapy (HRT) would reduce the risk of coronary heart disease. This still applies to apparently fit and healthy women but data from the Heart and Estrogen/progestin Replacement Study (HERS) have shown that the use of HRT in women with established risk factors for coronary heart disease increases mortality in the short term. A protective effect of HRT on the risk of coronary heart disease was not observed until after 2-3 years of treatment. Patient access to medical information has been revolutionized during the last decade. The availability of books, magazines and newspaper articles, which have always been an important source of patient information, has greatly increased. In addition, the Internet has transformed patient knowledge. According to surveys, these forms of communication provide the major source of information for 50% of women seeking advice about HRT. Indeed, the patient is often aware of the latest medical information before her doctor, possibly because she has more time. With such a large readership, these forms of communication have to report accurately. Unfortunately, they frequently do not and errors in reporting change the emphasis of a piece of medical research. The result is women are frightened and continuance suffers. It is worth remembering that 'bad news', suitably publicized, sells more newspapers and magazines than 'good news'. Competition is growing for 'alternative' products. 'Natural' treatments are 'in'. These treatments are widely claimed to reduce menopausal complaints in older patients and patients are keen to take them. But where is the evidence of efficacy and safety from robust and rigorous studies? All too often it is lacking. The doctor therefore has to advise the patient accurately about products for their treatment, and may also have to provide an opinion on 'alternative' remedies. Knowledge of alternative medical literature is therefore becoming essential. If doctors wish to continue to be respected by patients, they must remain knowledgeable and communicate their knowledge in an unbiased manner. They must emphasize the benefits and risks of different treatments according to the concerns of the patient. Failure of the doctor to provide this service will result not only in loss of respect, but alas, too often, in medico-legal consequences. PMID- 11379385 TI - The importance of patient education in improving compliance. AB - It is self-evident that the benefits of hormone replacement therapy will not be realized if women do not continue taking the therapy for long enough. All surveys show very disappointing levels of compliance or continuance, with typically about 25% of women stopping within 6 months and very few remaining on therapy for more than 1 or 2 years. The main reasons for stopping include unexpected or unacceptable side-effects, bleeding, fears of cancer, ignorance of the effects of hormone replacement therapy and the potential benefits, inappropriate expectations and poor support from medical advisers. With the ever-increasing options for prescribing hormone replacement therapy, including different routes of administration, combinations of hormones and variable dosages, it should be possible to find a regimen of hormone replacement therapy to suit most women. In the long term, poor continuation of hormone replacement therapy is strongly influenced by the presence of bleeding. Continuous combined estrogen/progestogen regimens, which avoid cyclical bleeding, are more acceptable, particularly for the older postmenopausal woman. However, the initial counselling has a most important impact and time taken to explain and to involve the woman in the decision process will encourage greater confidence in the merits of the therapy. The use of informative literature, videos and telephone help-lines, together with regular and close follow-up until a regimen is satisfactorily established, allows reassurance about initial side-effects or worries and adjustment of therapy. Recent developments of lower-dose combination therapy, selective estrogen receptor modulators and intrauterine progestogen-releasing systems should further improve compliance, but the key to continuation with hormone replacement therapy is a well-informed and confident patient. PMID- 11379386 TI - Disposition kinetics of florfenicol in goats by using two analytical methods. AB - Florfenicol, a monofluorinated analogue of thiamphenicol, has antibacterial activity against a broad spectrum of bacterial strains, including enteric bacteria that are resistant to chloramphenicol and thiamphenicol. The pharmacokinetics of florfenicol was studied following a single intravenous bolus or intramuscular injections at a dose of 20 mg/kg body weight, in five healthy goats. Serum florfenicol concentrations were determined using two analytical methods: microbiological assay and high-performance liquid chromatography (HPLC). Pharmacokinetic analysis was performed using redundant routine equations and the results derived from each method were compared. While florfenicol was detected for up to 4 and 8 h after administration by the bioassay, the drug was recovered in serum after 12 and 24 h by HPLC following intravenous and intramuscular injections, respectively. Comparison of the concentration profiles obtained by the two methods revealed substantial differences in the resultant kinetic data. Values for the initial serum concentration, elimination half-life, the area under the serum concentration-time curve, the mean residence time, and the systemic bioavailability were significantly (P < 0.01) higher when florfenicol concentrations were determined using HPLC. In conclusion, differences between analytical methodologies should be considered when interpreting the kinetic data for clinical use. However, both the hepatic biotransformations and the interchangeability of enantiomers need further investigation. PMID- 11379387 TI - Field evaluation of a bioregulator containing live Bacillus cereus spores on health status and performance of sows and their litters. AB - The efficacy of Paciflor, a bioregulator containing live Bacillus cereus CIP 5832 spores, was assessed in sows during late pregnancy and lactation, as well as in their piglets up to the growing phase. Two groups each of 30 pregnant gilts and sows received normal feed (T1 group), or feed with 85 g Paciflor per ton feed (T2 group), from 15 days prior to farrowing up to the end of the lactation period. Furthermore, 15 litters of the T1 group and 15 litters of the T2 group, were offered normal feed from the 5th to the 70th days of life (T1.1 and T2.1 groups, respectively), while the remaining 15 litters each of the T1 and T2 groups received the same feed but including Paciflor at a dose of 100 g/ton (from day 5 to day 49) and 50 g/ton (from day 50 to day 70). These pig litters were T1.2 and T2.2, respectively. No differences were seen between the T1 and T2 groups with respect to the clinical observations (loss of appetite, fever, mastitis, metritis and returns to oestrus, treatments applied, deaths, or removals to the slaughterhouse), gestation length, bodyweight of sows at farrowing or litter-size at birth. However, during lactation, the fat content of the dam's milk was increased (0.46% more fat), the body weight loss of sows was reduced and the number of weaned pigs per sow was increased (0.6 more pigs per litter) after administration of Paciflor (P < 0.05). Weaning to service interval was also reduced by 1 day (P < 0.05). Moreover, piglets receiving Paciflor with their feed (T1.2 and T2.2 groups) showed less incidence of scours and lower mortality compared to the untreated piglets (T1.1 and T2.1 groups), particularly those pigs originating from Paciflor-treated dams (T2.2 group) (P < 0.05). Despite the fact that no difference was seen between groups with regard to the amount of feed consumed, the feed conversion ratio of Paciflor-treated piglets (T2.2 and T1.2) was significantly improved compared to that of the untreated piglets (T2.1 and T1.1) (P < 0.05). With respect to weight gain, for the Paciflor-treated piglets, those born to Paciflor-treated mothers (T2.2) were 0.56 kg heavier than those born to untreated dams (T1.2) (P < 0.05). It is concluded that administration of Paciflor in dams during the end of pregnancy and during lactation, as well as to their offspring during suckling and the flat-deck period is beneficial for the survival and growth of the piglets. PMID- 11379388 TI - In vitro hepatobiotransformation of sulphadimethoxine in laying hens. AB - The biotransformation of sulphadimethoxine (SDM) was estimated in liver post mitochondrial supernatants (S-9) from laying hens. The pathway and activity for hen S-9 were compared with those for cow and pig S-9. The formation of the hydroxylation of SDM, 6-hydroxy SDM (6-OH-SDM), was found only with hen S-9. The N4-acetylating activity of SDM in hen S-9 was lower than in cow and pig S-9. All S-9 from the three species de-acetylated N4-acetyl SDM (N4-AcSDM). With respect to the hydroxylation and N4-acetylation rates in hen S-9, the values incubated at 41 degrees C were higher than those at 37 degrees C (P < 0.01). PMID- 11379389 TI - Effects of indomethacin on Salmonella typhimurium- and cholera toxin-induced fluid accumulation in the porcine small intestine. AB - The effect of the cyclooxygenase and prostaglandin E2 (PGE2) synthesis inhibitor, indomethacin, on the secretory responses induced by Salmonella serotype Typhimurium (ST) and cholera toxin (CT), in the porcine small intestine was investigated. ST (10(10) colony-forming units) and CT (56 micrograms) were instilled in tied-off intestinal loops in young anaesthetized pigs receiving intravenous indomethacin in a total dose of 7.5 mg/kg, or saline. The accumulated fluid in the loops and the luminal content of endogenous secretagogues PGE2 and 5 hydroxytryptamine (5-HT) were measured. ST induced fluid accumulation in the jejunum, whereas CT induced fluid accumulation in the jejunum and ileum. Indomethacin had no effect on the secretory responses. Indomethacin had a significant effect on the luminal content of PGE2 in jejunal ST and CT loops, whereas no effect of indomethacin was observed on the luminal content of 5-HT in ST and CT loops. In ST and CT loops, an increased content of PGE2 and 5-HT compared with test loops infused with Ringer's solution was observed. These results indicate that the porcine jejunal secretory response to ST and CT does not involve prostaglandins although indomethacin has an influence on the luminal release of PGE2 but not of 5-HT. PMID- 11379390 TI - Vitamin C plasma concentrations and leg weakness in the forelegs of growing pigs. AB - Four litters (41 pigs) of cross-bred pigs were studied from 6 to 26 weeks of age. Blood samples were collected at 6, 13, 21 and 26 weeks of age and analysed for contents of vitamin C, calcium (Ca), inorganic phosphorus (P) and alkaline phosphatase (ALP). The pigs were examined clinically for foreleg weakness at the ages of 21 and 26 weeks. At the age of 26 weeks the pigs were slaughtered and the right forelegs were examined macroscopically and selected samples were collected for radiological, histological and ultrastructural examination. The prevalence of foreleg lesions was high, with lesions of dyschondroplasia of the distal growth plate of the ulna in 30 pigs, synovitis of the elbow joint in 24 pigs and osteochondritis dissecans of the elbow joint in 25 pigs. At the ages of 21 and 26 weeks, five pigs had evidently crooked forelegs and 14 pigs (age 21 weeks) and 25 pigs (age 26 weeks) had mildly deformed forelegs. The serum levels of Ca, P and ALP were within normal values for growing-finishing pigs. The range of vitamin C concentrations in plasma showed a wide difference (7.1-49.8 mumol/l) but was not associated with deformed forelegs. The serum concentrations of Ca, P and ALP and the plasma concentration of vitamin C differed significantly (P = 0.05) between age groups and there was a significant (P = 0.001) positive correlation between the levels of vitamin C in plasma and the serum levels of ALP at 6 weeks of age. The aim of the present study was to determine if there was any association between the plasma levels of vitamin C and the extent of crooked or deviated forelegs in growing-finishing pigs. We could not find a vitamin C deficiency during the study and no association between low levels of vitamin C in plasma and the presence of deformed forelegs of these 40 pigs. PMID- 11379391 TI - Activities of gamma-glutamyltransferase, alkaline phosphatase and aspartate aminotransferase in colostrum, milk and blood plasma of calves fed first colostrum at 0-2, 6-7, 12-13 and 24-25 h after birth. AB - Enzyme activities of gamma-glutamyltransferase (gamma-GT), alkaline phosphatase (AP) and aspartate-aminotransferase (AST) were measured from birth to the age of 28 days in calves which were fed colostrum at 0-2, 6-7, 12-13, or 24-25 h after birth. Enzyme activities were also measured in colostrum (first to fifth milking) and in mature milk. Activities were highest in the first colostrum milking and decreased to the lowest activities in mature milk. Plasma gamma-GT activity transiently increased after first colostrum intake and was greater in calves fed first colostrum within less than 6-7 h than in those fed first colostrum later than 12 h after birth. Activity of gamma-GT reflected the absorption of colostral gamma-GT, which decreased with time after birth. The AP activity transiently increased after colostrum intake and was higher in calves fed colostrum within the first 12 h of life than in those fed later after birth. The transient rise of plasma AP activity also indicated absorption of colostral AP, although endogenous sources of AP could not be excluded. The activity of AST also transiently increased after colostrum intake but there was no association with time of first colostrum feeding, indicating that the rise of plasma AST activity was the consequence of enhanced endogenous production and was independent of colostrum intake. In conclusion, there are different causes leading to postnatal changes in enzyme activities. PMID- 11379392 TI - Pharmacokinetic behaviour of phenylglyoxal bis(guanylhydrazone) (PGBG) after intravenous administration in rabbits. AB - Phenylglyoxal bis(guanylhydrazone) (PGBG) is a synthesized analogue of methylglyoxal bis(guanylhydrazone) (MGBG), which has demonstrated anti-parasitic activity in rabbits. The pharmacokinetic behaviour of PGBG after intravenous administration (10 mg/kg bodyweight) was studied in five rabbits. Plasma concentrations of PGBG were measured by high-performance liquid chromatography. Plasma PGBG concentrations decreased rapidly and were not detectable beyond 90 min after treatment. The mean [+/- standard deviation (SD)] volume of distribution at steady state (Vdss) was 2.19 +/- 0.47 l/kg and the mean plasma clearance value (Cl) was 29.99 +/- 3.98 ml/min kg. This drug is rapidly eliminated from the body in rabbits, having a short elimination half-life (0.93 h) and mean residence time (1.21 h). PMID- 11379393 TI - [Brain tumor or stroke?]. AB - Sometimes, the clinical presentation of a brain tumour mimics that of stroke or vice versa, as exemplified in the following three patients. In a 73-year-old patient the initial clinical picture was compatible with a brachial plexus lesion, as the weakness in his right hand appeared to have a traumatic, and not a central nervous system related, cause. When he experienced a focal seizure, the CT scan of the brain revealed a lesion in the motor cortex. This was presumed to be an infarction due to the lack of mass effect and the absence of contrast enhancement. Shortly afterwards the patient deteriorated and a follow-up scan revealed a large contrast-enhancing lesion. During surgery this proved to be a glioblastoma multiforme. A 76-year-old man was suffering from a progressive neurological deficit. An MRI scan of the brain revealed a contrast-enhancing lesion and a chest X-ray revealed an asymptomatic lung tumour; the diagnosis 'brain metastasis' was made. The surgeon removed the lung tumour, which proved to be a carcinoma. Later, when the patient was referred to the neurosurgeon for extirpation of the presumed brain metastasis, the MRI scan revealed that the lesion had decreased in size and no longer exhibited contrast enhancement. The metastasis proved to be an infarction. A 53-year-old man presented with sudden loss of consciousness due to a haemorrhage in the occipital lobe. An angiogram did not reveal a vascular malformation and during surgery no abnormal tissue was seen. The patient almost made a complete recovery. However, several months later he developed an elevated intracranial pressure due to a large occipital high grade glioma, which had caused the original haemorrhage. PMID- 11379394 TI - [Stereotaxic neurosurgery for treatment of Parkinson disease]. AB - Three target structures are available in stereotactic surgery for Parkinson's disease: the thalamus, the globus pallidus and the subthalamic nucleus. The subthalamic nucleus appears to be the most promising structure. However, the thalamus can be considered in the case of an incapacitating tremor presenting as a primary symptom. Surgery in the globus pallidus may be as effective as in the subthalamic nucleus, but in the latter it is often accompanied by a reduction in dopaminergic medication. The surgical technique of electrical stimulation causes fewer adverse effects than that of coagulation and can therefore be applied bilaterally, but does require more intense postoperative care. In the selection of patients for surgery, levodopa responsiveness plays an important role in predicting effectiveness, except in the case of tremor. PMID- 11379395 TI - [Spiral computed tomographic angiography as a substitute for intra-arterial angiography of aorta and its branches]. AB - Until recently, intra-arterial angiography was the diagnostic method of first choice when pathology of the aorta or its branches was suspected. A disadvantage of this technique is that only the lumen of spaces with blood flow can be visualised and that the soft tissue surroundings remain (partly) invisible. Spiral computer tomographic angiography (CTA) has some major advantages compared with conventional angiography. The technique is less invasive and faster. Also, the soft tissue is imaged by CTA. In addition, computer reconstructions allow viewing from all directions without the limitations of overprojection. Spiral CTA is a suitable technique for imaging the thoracic part of the aorta: in case of dissection if transoesophageal echography is not available, in case of an aneurysm to determine the diameter and in case of rupture as a highly sensitive but not very specific examination technique. For imaging of the abdominal part of the aorta, spiral CTA may be considered. In case of an aneurysm or a possible rupture of this part of the aorta it is then possible to visualize the operation area and to choose the optimal approach. For the exclusion of stenoses in mesenteric arteries or in renal arteries, spiral CTA offers the advantage of non invasivity. The technique is less suitable for demonstration of these stenoses and does not allow immediate intervention. PMID- 11379396 TI - [Diagnostics for classification of anemia]. AB - Examining peripheral blood smears provides valuable information in the diagnosis of anaemia despite large inter- and intraobserver variation. The classification of anaemia is usually based on the average erythrocyte size, referred to as the mean corpuscular volume (MCV). Microcytosis indicates a reduced haemoglobin synthesis caused by either an iron deficiency or haemoglobinopathy, a congenital disorder. Macrocytosis is the result of a disruption to the division and maturing of proerythroblasts in the bone marrow, due, for example, to vitamin B12 (folic acid) deficiency or excessive alcohol use. Furthermore, a high number of reticulocytes in the blood indicates an increased production of erythrocytes whereas a low total indicates an inadequate production level. In addition to the case history and the physical examination, the MCV and number of reticulocytes can provide guidance with respect to further diagnostic investigation. PMID- 11379397 TI - [From gene to disease; thyroid stimulating hormone receptor, hyperthyroidism and hypothyroidism]. AB - The thyroid-stimulating hormone receptor (TSH-R) gene appears to be very sensitive to mutagenesis, in view of the vast number of reported mutations. Loss of-function germline mutations occur preferentially in the hormone-binding extracellular domain of the TSH-R, resulting in familial TSH resistance. Gain-of function germline mutations occur preferentially in the transmembrane domain of the TSH-R, resulting in familial non-autoimmune hyperthyroidism. Familial gestational hyperthyroidism is due to a mutant TSH-R which is hypersensitive to chorionic gonadotropin. Somatic gain-of-function mutations are a major cause of toxic thyroid adenomas. PMID- 11379398 TI - [Diagnostic image (36). Idiopathic scrotal edema]. AB - In a 5-year-old boy idiopathic scrotal edema was diagnosed, a rare self-limiting disease. PMID- 11379399 TI - [Successful reanimation of facial paralysis with an indirect anastomosis between hypoglossal nerve and facial nerve, without loss of function of the tongue]. AB - OBJECTIVE: To describe the surgical procedure and the results of the indirect hypoglossal-facial nerve anastomosis using a free nerve graft in patients with facial nerve paralysis. This technique leaves the tongue function intact. DESIGN: Prospective study. METHODS: Tongue function was assessed in 39 consecutive patients who underwent this procedure and facial reanimation was assessed in 29 of these patients who had completed at least 24 months follow-up. Facial nerve function was judged using the House-Brackmann (HB) facial nerve grading system. RESULTS: Tongue movements were normal in all operated patients; one patient had mild homolateral atrophy. Initial facial movements occurred on average 7.5 months postoperatively (range 4 to 18 months) in all but one patient. The results were graded HB II in 6 patients (20.9%), HB III in 13 (44.6%), HB IV in 7 (24.1%), HB V in 2 (6.8%) and HB VI in 1 patient (3.4%). Hemifacial synkineses were noticeable but no mass movements or gross hypertonia were observed (as are often present in direct hypoglossal-facial anastomosis). The results of facial reanimation were significantly better in young patients and when a short time interval between paralysis and surgery existed. HB grade II was achieved only if the duration of paralysis was less than 12 months. CONCLUSION: Indirect hypoglossal-facial anastomosis with interposition of a nerve graft allows preservation of tongue function together with good overall facial reanimation, and is therefore to be preferred to the classical direct hypoglossal-facial anastomosis. PMID- 11379400 TI - [Decline in the need for blood transfusions in cancer patients due to the use of epoietin alfa during cisplatin based chemotherapy]. AB - OBJECTIVE: To compare the need for blood transfusions between two groups of patients treated with cisplatin-containing chemotherapy. One of these groups received epoetin alpha therapy. DESIGN: Prospective with an historical control group. METHODS: From April 1998 to December 1999, 44 patients who were being treated with cisplatin and gemcitabine were also administered epoetin alpha (10,000 U subcutaneously, thrice weekly) from the onset of anaemia. The need for red blood cell transfusions in this group was compared to a historical control group of 46 patients treated with the same combination chemotherapy regimen from November 1995 to July 1997. RESULTS: In the historical control group, each patient received an average of 1.6 red blood cell transfusions as compared to 0.7 in the epoetin alpha group (a 58% reduction). The average number of units of red blood cells transfused per patient was 3.6 for the control group and 1.8 for the epoetin alpha group (a 50% reduction). In the epoetin alpha group, none of the patients received more than 2 transfusions whereas in the control group, 10 patients (22%) received 3 or more transfusions. In two patients, epoetin therapy had to be stopped due to the occurrence of hypertension. CONCLUSION: Epoetin alpha reduced the need for red blood cell transfusions during cisplatin containing chemotherapy. Its toxic effect was minimal. PMID- 11379401 TI - [Chronic pain and impairment of function after a sting by the great weaver fish (Trachinus draco)]. AB - A 50-year-old fisherman was stung in his right hand by a Great Weever fish (Trachinus draco). The crew did not have sufficient medical knowledge to adequately treat him on board. Severe pain, oedema of the hand, fever, vomiting and syncope occurred. Treatment with antibiotics, on board, after disembarkation and later in hospital for six days reduced the severity of the symptoms. However, two years after the accident, the patient still suffered from a dysfunction of the right hand as well as extreme fatigue and intermittent joint complaints. The symptoms mostly commonly arising from a Weever fish sting are: severe pain, local erythema and oedema. Systemic symptoms may sometimes occur: headache, syncope, bradycardia, fever and hypotension. The symptoms of continuous joint pain and severe fatigue following a Weever sting have not been previously described. The Weever fish venom contains a mixture of biogenous amines, of which some are known: 5-hydroxytryptamine, epinephrine, norepinephrine and histamine. The venom's composition has yet to be fully elucidated. In the event of a Weever fish sting, the first aid which should be given is: clean the wound and immerse the affected part of the body for at least 30 minutes in water which is as hot as the victim can tolerate (40-45 degrees C). Persons at risk from Weever fish stings are bathers, especially from the Lesser Weever fish (Echiichthys vipera), and sea fishermen. General practitioners and first aiders in coastal areas as well as sea fishermen should be informed about the first aid to be given in the event of a Weever sting. PMID- 11379402 TI - [Embolization and treatment for post partum hemorrhage]. PMID- 11379403 TI - [Lung cancer in Netherlands during the period of 1989-1997: the epidemic is not over yet]. PMID- 11379404 TI - [Physical diagnosis--ascites]. PMID- 11379405 TI - 2nd Chapter of Paediatricians Lecture: the future of paediatrics in Singapore. PMID- 11379407 TI - A case series of six children with primary pulmonary hypertension. AB - INTRODUCTION: Primary pulmonary hypertension is an uncommon but serious disease in children. Management is difficult despite recent advances in pharmacotherapy. CLINICAL PICTURE: We reviewed patients with this condition with respect to their presenting symptoms, investigations, treatment and outcome. TREATMENT: These children were treated with individualised combinations of oxygen, diuretics and calcium antagonists. OUTCOME: In our follow-up of 5 children, 4 had died. CONCLUSION: This condition is irreversible and progressive with a high mortality rate. A better understanding of, and research into, the pathogenesis would hopefully lead to the formulation of improved therapeutic strategies for this condition. PMID- 11379406 TI - Renal dysfunction in workers exposed to inorganic lead. AB - INTRODUCTION: We studied the relationship between renal dysfunction and occupational lead exposure in a local factory. MATERIALS AND METHODS: A cross sectional study with a cohort component was conducted with 55 male workers of a factory producing PVC stabilisers as subjects. Workers from this factory have been followed up with 6-monthly blood lead measurements since 1982. Two indices of overall lead exposure, i.e. the number of times a worker's blood lead exceeded 40 micrograms/dL (PbB40) and cumulative blood lead index (PbBint), were obtained from this data. Recent blood lead concentration (PbB) was measured. 4-hour creatinine clearance and various other urinary and serum markers of renal dysfunction were used as effect indices. RESULTS: There was no relationship between PbB and any of the renal markers. However, creatinine clearance decreased significantly (P < 0.001) with increasing PbB40 and PbBint after adjustment for age and smoking habits. Urinary albumin (Ualb), urinary alpha-1 microglobulin (U alpha 1m), urinary beta-2 microglobulin (U beta 2m) and urinary retinol-binding protein (URBP) increased significantly with both increasing PbB40 and PbBint. Total urinary activity of N-acetyl-beta-D-glucosaminidase (NAG-T) and its heat stable isoenzyme (NAG-B) increased significantly with increasing PbB40. A significant difference in renal parameters occurred when PbB40 was 1 or more. CONCLUSIONS: We have found a positive association between overall lead exposure and renal dysfunction. The renal parameters were significantly higher among those who had at least one episode of blood lead above 40 micrograms/dL. Our findings also strengthen the case for the use of Ualb, U alpha 1m, U beta 2m, URBP, NAG-T and NAG-B as early markers of lead nephropathy. PMID- 11379408 TI - Trinucleotide repeat analysis of Huntington's disease gene in Singapore. AB - INTRODUCTION: Huntington's disease (HD) is an inherited neurodegenerative disorder characterised by chorea and progressive dementia. The mutation causing the disease has been identified as an unstable expansion of a trinucleotide (CAG)n. We have assessed the (CAG)n repeats in the patients and controls in our population. MATERIALS AND METHODS: Polymerase chain reactions (PCRs) for the repeat region were carried out for 116 individuals: 10 were asymptomatic at-risk members from 5 families; 53 symptomatic patients from various hospitals; and 53 normal unrelated Singaporeans. Estimation of the number of repeats was based on Metaphor gel electrophoresis, sizing using the GeneScan on ABI 310 Genetic Analyzer, and sequencing using the same equipment. RESULTS: Metaphor gel sizing generally gives an over-estimation, and GeneScan gives an under-estimation of repeat numbers compared with sequencing which is the gold standard. Of the 63 patients and family members tested, 25 had one expanded allele of 40 to 54 CAG repeats and the other allele in the normal range of 15 to 30 repeats. One patient had an allele in the intermediate range (38). CONCLUSION: The range of CAG repeats in the normal and HD alleles in our population is similar to those reported elsewhere. An accurate sizing can only be obtained with sequencing. For allele sizes in the intermediate range (37-40), sequencing should be carried out to confirm the carrier status of a patient. PMID- 11379409 TI - Current and emerging treatments in Parkinson's disease. AB - INTRODUCTION: Parkinson's disease is one of the commonest neurodegenerative diseases of the elderly. The discovery of dopamine deficiency from the degeneration of substantia nigra neurons revolutionised treatment in the early 70's. METHOD: This review is based on Medline search with Keywords (Parkinson's disease, treatment, therapy, surgery). RESULTS: Levodopa remains the gold standard of medications. New drugs include recently introduced dopamine agonists like ropinorole and pramipexole, and COMT inhibitors. The role of selegiline in neuroprotection remains controversial. Dopamine agonist monotherapy may delay the onset of motor fluctuations. Levodopa may be associated with the earlier onset of motor complications such as dyskinesias, and it may be preferable to delay its use in younger patients. In older patients, especially those with significant disability, levodopa use should not be delayed. Surgical therapies such as pallidotomy, bilateral subthalamic deep brain stimulation and foetal transplantation can be considered for those who fail medical therapy. Molecular science techniques including gene therapy, neurotrophic factors, stem cell technology form the next frontier of Parkinson's research. CONCLUSION: No new medication has proven more efficacious than levodopa. More controlled trials are required in Parkinson's surgery to reach definite conclusions about its effects and long-term results. PMID- 11379410 TI - Neuroprotection in acute stroke. AB - PURPOSE: To highlight recent advances in neuroprotection in acute stroke. DATA SOURCES: A MEDLINE search was conducted from January 1985 to November 2000. Key words included neuroprotection, cerebrovascular, subarachnoid haemorrhage, perioperative stroke, hypothermia and apopotosis. All articles in English were considered for review. Additional articles were identified from the references of the retrieved articles and cross-referencing selected articles. DATA EXTRACTION: All clinical studies and review articles and abstracts were reviewed. DATA SYNTHESIS: The neuronal cells of the central nervous system are susceptible to various forms of insult such as ischaemia and haemorrhage. Each step along the ischaemic cascade is a potential target for therapeutic intervention. Neuroprotective agents are designed to minimise cellular injury and salvage brain tissue. In cerebral ischaemia, only thrombolysis had been shown to improve clinical outcome. Neuroprotective therapy has definite benefits in animals but not in humans. It may potentially extend the time window for thrombolysis. In aneurysmal subarachnoid haemorrhage, the only agent with proven efficacy is nimodipine. Research is ongoing in the development of new drugs. Currently several phase III trials are in progress. CONCLUSION: There is substantial optimism in the development of neuroprotective therapy to improve outcome in stroke patients. PMID- 11379411 TI - The eye in neurology: evaluation of sudden visual loss and diplopia--diagnostic pointers and pitfalls. AB - The evaluation of sudden visual loss should begin with the differentiation between monocular loss and binocular loss. The importance of this is reflected in the differences between the main causes of monocular and binocular losses. In cases of transient monocular visual loss, an ocular cause has to be kept in mind so as to avoid unnecessary and costly cerebrovascular investigations. In cases of persistent monocular visual loss, a compressive lesion of the optic nerve or chiasma may simulate optic neuritis. In the evaluation of diplopia, the main differential diagnoses are nerve lesions and myasthenia. The main causes of nerve lesions responsible for diplopia and their workup are summarised. The usefulness of eye signs in the diagnosis of myasthenia is highlighted. The possibility of compressive lesions co-existing with or masquerading as myasthenia is emphasised. PMID- 11379412 TI - The impact of swallowing disorders in the elderly. AB - INTRODUCTION: Swallowing disorders are common in the elderly but its prevalence is often underestimated. They can result in increased morbidity and mortality. METHODS: This article summarises the findings of selected published papers in major international journals indexed on Medline on swallowing using the key words -swallowing, dysphagia, aged, geriatrics and deglutition. RESULTS: There are age related changes in the oral, pharyngeal and oesophageal functions. In the elderly, central nervous system diseases such as stroke, parkinsonism, dementia, medications, local oral and oesophageal factors are common causes of swallowing dysfunction. Swallowing disorders in the elderly are associated with increased mortality and morbidity. Aspiration, dehydration, pneumonia, malnutrition, functional decline and institutionalisation are often encountered in the elderly with dysphagia. There is a choice of different interventions available to reduce morbidity and mortality arising from swallowing impairments, and improving their quality of life. CONCLUSION: The effective management of swallowing impairment in the elderly requires a multidisciplinary team approach. PMID- 11379413 TI - Behavioural disorders in childhood: a Singapore perspective. AB - INTRODUCTION: Behaviour disturbances in preadolescent children are common, ranging from mild disturbances to full disorders meeting official nosological criterion. The objective of this paper was to review behavioural disorders occurring in the preadolescent child using up-to-date published data supplemented by available local published data on the subject. METHODS: Recent reviews of the various psychological disorders in childhood that have a significant behavioural component were identified through a MEDLINE search (from 1990). Information from these reviews was supplemented with relevant local studies cited on MEDLINE from 1990. Information on common behavioural disturbances facing parents locally was obtained from questions submitted to the author over a 3-year period by a local parenting magazine. A search was also conducted on the DOW JONES publications archives for relevant information from the local news media. RESULTS: The perception of a problem may arise from the child, family, school, community or society's perspective. Behavioural problems can be conceptualised to exist in the following broad categories: Behaviour problems/disorders related to daily physiological activities; disruptive behavioural problems and disorders; pervasive developmental disorders and disorders of social behaviour; behaviour problems related to emotional disorders; and behaviour problems associated with other mental conditions in children. Family, behavioural and multisystemic interventions are the commonest non-drug interventions. Drug use is on the increase, especially in disruptive, affective and anxiety disorders. CONCLUSIONS: Behavioural disorders in children are common. They are often under-recognised and under-treated. Improved recognition demands that a good assessment is being carried out and a heightened index of suspicion for the commoner behavioural disorders. Adopting an evidence-based medicine approach and utilising recent advances in psychopharmacology and psychotherapy may improve outcome. PMID- 11379414 TI - New cyclooxygenase inhibitors. AB - INTRODUCTION: The discovery of the two isoforms of cyclooxygenase (COX-1 and COX 2) has paved the way for the development of a new class of non-steroidal anti inflammatory drugs (NSAIDs). The COX-2 inhibitors have shown comparable efficacy to the traditional NSAIDs with less gastrointestinal side effects in major clinical studies. The aims of this paper are to provide a brief historical background, pharmacology of cyclooxygenase inhibitors as well as discuss the latest update on COX-2 inhibitors. METHODS: A MEDLINE search was performed for relevant articles from leading medical journals from 1990 to 2000. The papers reviewed include randomised controlled clinical trials, meta-analyses and review articles. CONCLUSION: The arrival of the COX-2 inhibitors into the treatment armamentarium for inflammation and analgesia has been widely accepted and welcomed. Although this new group of "wonder" drugs cause less gastrointestinal side effects, they are not renal sparing and do not have cardioprotective effects. Studies have also yet to establish its safety in NSAID-sensitive patients. In patients with chronic arthritides, cost is also an important factor to consider when prescribing these medications. Their strengths and weaknesses will become apparent with continued use. PMID- 11379416 TI - Enteral nutrition of the very low birth weight (VLBW) infant. AB - INTRODUCTION: Optimal nutrition is critical in the management of the very low birth weight (VLBW) infant. METHODS: The type of feeding, initiation, route and schedule of feeding, and the rate of increase in feeding volume were reviewed. RESULTS: Human milk from the preterm infant's mother is the feeding of choice. When full enteral feeding is established, supplementation of human milk with a multi-nutrient fortifier is required. In VLBW infants having poor weight gain when exclusively fed human milk, feeding of hindmilk is encouraged if the mother expresses large quantities of breast milk. Lactation support is vital for the mother, including appropriate methods of milk collection, storage and transport to the nursery. In mothers who produce no or insufficient human milk, preterm formulas are an alternative feeding, used either alone or in conjunction with human milk. Minimal enteral nutrition using sub-nutritional feedings in the first week of life is advantageous. Intragastric intermittent ("bolus") tube feeding is the route of choice for the infant born at < 32 to 34 weeks gestational age. Feeding volume must be advanced cautiously at < 20 mL/kg/day. CONCLUSION: Future goals of nutrition research in the VLBW infant must include determining the role and optimal composition of the various nutrients and duration of use. PMID- 11379415 TI - New disease modifying agents in adult rheumatoid arthritis. AB - In recent years, new disease modifying agents including leflunomide and tumour necrosis factor (TNF) antagonists have been used to treat patients with rheumatoid arthritis (RA). Leflunomide prevents proliferation of activated lymphocytes by inhibiting dihydroorotate dehydrogenase, a critical step in de novo pyrimidine synthesis. Leflunomide has been shown to be as effective as sulfasalazine and methotrexate (MTX) in placebo-controlled trials. It also improves physical function, quality of life measures and retards radiographic progression. TNF antagonists include infliximab and etanercept. Infliximab is a chimeric TNF monoclonal antibody. Repeated infusions of low dose infliximab (1 mg/kg) are ineffective if given alone. Addition of MTX to infliximab has been shown to prolong the duration of clinical response. Etanercept is a human TNF receptor p75 Fc fusion protein. In active RA patients with suboptimal response to MTX, additional clinical benefit was obtained by the addition of infliximab or etanercept to MTX. The main side effect of etanercept is injection site reaction. However, the long-term effect of TNF antagonists in the development of infection, malignancy and autoimmune disease remains unknown. PMID- 11379417 TI - An introduction to biodegradable materials for tissue engineering applications. AB - Tissue generation by autogenous cell transplantation is one of the most promising treatment concepts being developed as it eliminates problems of donor site scarcity, immune rejection and pathogen transfer. Cultured cells are seeded onto a three-dimensional biocompatible scaffold that will slowly degrade and resorb as the soft and hard structures grow and assimilate in vitro and/or in vivo. The 3-D scaffold provides the necessary template for cells to proliferate and maintain their differentiated state. Ultimately, it defines the overall shape of the tissue-engineered transplant. The aim of this review is to describe and discuss the scaffold materials of natural and synthetic origin that are of specific interest to tissue engineers. This review is based on previous publications and our own experience in the use of biomaterials of natural and synthetic origin for tissue engineering applications. Biodegradable polymers which have been used for tissue engineering applications are mainly based on clinically established medical devices and implants. In the group of macromolecules of natural origin collagen, alginate, agarose, hyaluronic acid derivatives, chitosan, and fibrin glue have been used as scaffolds. Man-made polymers such as polyglycolide (PGA), polylactides (PLLA, PDLA), poly(caprolactone) (PCL), and poly(dioxanone) (PDS) have been studied as matrix material to guide the differentiation and proliferation of cells into the targeted functional premature and/or mature tissue. Appropriate selection of scaffold material with respect to the targeted tissue is essential. Today, biomaterials of choice remain to be those approved by the US Food and Drug Administration. In spite of that, novel biomaterials should be developed specifically designed for tissue engineering applications. PMID- 11379418 TI - Pulse dye densitometry: a novel bedside monitor of circulating blood volume. AB - INTRODUCTION: Monitoring of circulating blood volume is important in the management of critically ill patients. Current methods of circulating blood volume measurements such as indicator dilution using radioisotopes or Evans blue dye are unsuitable for clinical application as these tests do not allow for frequent repeated measurements to be done. A direct bedside measurement of circulating blood volume using the principle of pulse dye densitometry was recently introduced. This is essentially an indicator dilution technique using indocyanine green combined with the principle of pulse spectrophotometry. METHODS: This paper aims to review this method of circulating blood volume measurement and provide a summary of the published clinical trials that compared its accuracy with the other conventional methods of circulating blood volume measurement, based on a Medline search, spanning the period 1966 to August 2000. RESULTS: Published studies show that pulse dye densitometry gives comparable results when compared to other conventional methods of blood volume measurement. Its ability to measure circulating blood volume accurately and repeatedly, as frequently as every 20 min makes it suitable for clinical application. CONCLUSION: Pulse dye densitometry provides for a rapid, semi-noninvasive and convenient bedside assessment of circulating blood volume that is applicable clinically. Further studies are needed to ascertain the impact of the use of pulse dye densitometry on the mortality and morbidity of the critically ill. PMID- 11379419 TI - Bugs for the next century: the issue of antibiotic resistance. AB - OBJECTIVE: To address the issue of emerging antibiotic resistance and examine which organisms will continue to pose problems in the new century. METHODS: Review of articles pertaining to bacteria recognised for increasing resistance. RESULTS: Changing resistance patterns are correlated with patterns of antibiotic use. This results in fewer effective drugs against "old" established bacteria e.g. gram-positives such as Streptococcus pneumoniae and Staphylococcus aureus. Resistance in gram-negative bacteria is also steadily increasing. Nosocomial gram negative bacteria are capable of many different resistance mechanisms, often rendering them multiply-resistant. Antibiotic resistance results in morbidity and mortality from treatment failures and increased health care costs. CONCLUSION: Despite extensive research and enormous resources spent, the pace of drug development has not kept up with the development of resistance. As resistance spreads, involving more and more organisms, there is concern that we may be nearing the end of the antimicrobial era. Measures that can and should be taken to counter this threat of antimicrobial resistance include co-ordinated surveillance, rational antibiotic usage, better compliance with infection control and greater use of vaccines. PMID- 11379420 TI - Primary anorectal malignant melanoma: clinical features and results of surgical therapy in Singapore--a case series. AB - INTRODUCTION: Primary malignant melanoma arising from the anorectum is uncommon. The natural history of anorectal malignant melanoma is that of a very poor prognosis with early dissemination of disease. Successful surgical treatment has been rare. The present series reviews the clinical features and results of surgical management of patients with anorectal malignant melanoma treated in the Department of Colorectal Surgery, Singapore General Hospital. MATERIALS AND METHODS: Data for all patients treated for anorectal malignant melanoma during an 11-year period from 1989 to 1999 were reviewed. The age, sex, presenting symptoms, duration of symptoms prior to diagnosis, size of tumour, extent of disease, type of surgery and length of survival were analysed. RESULTS: Four men and 2 women, ranging in age from 31 to 81 years with histologically proven primary anorectal malignant melanoma, were included in the study. The most common (67%) presenting symptom was rectal bleeding. The mean tumour size was 2.5 cm (range 1 to 5 cm). All underwent abdomino-perineal resection. Three died of disseminated disease within 17 months while the other 3 were still alive at the time of this study; the longest up to 6.5 years from the time of diagnosis. CONCLUSION: The prognosis of primary anorectal malignant melanoma is poor. However, it is worthwhile treating aggressively as long-term survivor may be encountered in some. PMID- 11379421 TI - Mucinous cystadenoma of the appendix--an unusual cause of intestinal obstruction. AB - INTRODUCTION: Report of a rare presentation of mucinous cystadenoma of the vermiform appendix. CLINICAL PICTURE: A lady who presented with intestinal obstruction and peritonitis was found to have gangrenous small bowel caused by strangulation by a tumour of the appendix. TREATMENT: Right hemicolectomy. OUTCOME: The histology of the appendicular tumour was mucinous cystadenoma. The patient recovered well. CONCLUSION: Mucinous cystadenoma of the appendix can present in many ways and it is important to recognise the pathology at operation. PMID- 11379422 TI - The Runme Shaw Memorial Lecture 2000--treating VIPs in Washington. PMID- 11379424 TI - 11th Chapter of Surgeons Lecture--the art of letting go. PMID- 11379423 TI - 15th Gordon Arthur Ransome Oration: a journey of care--mission impossible? PMID- 11379426 TI - 10th Seah Cheng Siang Memorial Lecture: going places--a rheumatological odyssey. PMID- 11379425 TI - 5th Yahya Cohen Lecture: angiosomes and extension of skin flaps--anatomic study and clinical implications. PMID- 11379427 TI - Prenatal treatment of congenital adrenal hyperplasia: author differs with technical report. PMID- 11379428 TI - Developing a practical guide to assess the potential health impact of urban regeneration schemes. AB - We report here on the development of a practical guide for health impact assessment (HIA) of urban regeneration schemes which sets out an approach to rapid prospective HIA. We provide a succinct account of our approach, illustrated by two case studies in East London, and discuss how the process of HIA must be adapted to different types of regeneration projects. The case studies are described in more detail in Cave and Curtis (2001). In this paper we focus on how the HIA process was adapted to suit different types of regeneration programmes. PMID- 11379429 TI - Promoting human impact assessment within the environmental impact assessment process: Canada's work in progress. PMID- 11379430 TI - Health impact assessment of chrome-based leather tanning: short report of an ongoing case study of Pakistan. PMID- 11379431 TI - Health promotion within the development process. AB - The aim of this article is to show how health promotion can play a more active and central role in the development process. In demonstrating this role, the skills necessary to deliver these new strategies will emerge which build on existing advocacy skills. PMID- 11379432 TI - Meanings of networking: a perspective. PMID- 11379433 TI - Time for renewal--once again. PMID- 11379434 TI - Instant messaging: a potential for virtual meetings in the health field. PMID- 11379435 TI - Targeting, tailoring and stages of change. AB - This is the first of an occasional series of articles that will feature in full exclusively in Promotion & Education as part of the formal link between IUHPE and Health Education Research. Each article will review key issues raised by papers published recently in Health Education Research on a topic or theme of current relevance to health promotion. A brief summary will also appear in Euronews, the IUHPE Regional Newsletter for Europe. PMID- 11379436 TI - Health impact assessment in Australia and New Zealand: an exploration of methodological concerns. PMID- 11379437 TI - Respiratory symptoms and ventilatory function of the sawmillers in Ibadan, Nigeria. AB - A total of five hundred sawmillers in Ibadan were studied to assess occupation induced lung impairment as a result of exposure to saw dust. Occupation-related symptoms were recorded using structured questionnaire. Age and sex matched controls consisting of 500 University College Hospital (UCH) Ibadan workers or students were used. They were apparently healthy and work and live at places free of fumes and smoke. Peak expiratory flow rate (PEFR) was measured in all subjects. However, full spirometry work-up was done on 120 sawmillers and 120 control subjects that had been selected using simple random sampling technique. The most frequent pulmonary symptoms among the sawmill workers were running nose and sneezing (57.40%) and productive cough (34.30%), while the symptoms of dyspnoea and wheezing were each present in (4.10%) of the subjects. The mean PEFR of the sawmillers (463.8 +/- 63.4 L/min) was significantly lower (P < 0.0001) than that of the control subjects (537.7 +/- 71.5 L/min). Similarly, the mean values of FEV1, FVC and FEV1% (FEV1%) were also significantly lower in the FVC sawmillers than the control subjects. The findings indicate that respiratory symptoms are common during the working hours among the sawmillers and 4.1% of the subjects studied suffered some degree of airway obstruction. PMID- 11379438 TI - Immune response to aflatoxin B1-histone H1 complex. AB - Antibodies against aflatoxin B1 have been raised in rabbits using aflatoxin B1 histone H1 conjugates as immunogen. Aflatoxin B1 was coupled to histone H1 via the ultimate carcinogen aflatoxin B1-2, 3-epoxide. The antibodies are specific for aflatoxin B1. The average number of binding sites on the antibody molecules for aflatoxin B1 as obtained from Scatchard plot analysis of the binding data is 1.94 with < F degree = -6.19 Kcal/mol, while the average association constant for the binding is 34.5 x 10(3) M-1. Male wistar rats after immunization with aflatoxin B1-histone H1 showed lower mortality and reduction of acute toxic effects in the liver when challenged with a single dose of aflatoxin B1. The antibodies may be useful in immunoprophylaxis against aflatoxicosis. PMID- 11379439 TI - Secretory IgA and complement levels in patients with hookworm infection in Zaria. AB - As a continuing investigation into the immunological reactions involved in hookworm infection the levels of secretory IgA (sIgA). IgM, IgA, complement C3 and C4 were studied in 57 Nigerians with hookworm infection and compared with those of 28 healthy, normal controls. The hookworm patients were divided into three groups based on the degree of anaemia (Hb1-7 and hypochromia +++). Group II had moderate anaemia (Hb8-11, hypochromia ++ and Group III had no signs of anaemia despite the underlying hookworm infection. The patients also comprised those in whom hookworm was the sole infection and those with hookworm associated with other parasites. Significant differences in the results between each patient subgroup and the controls were analysed using the student t-test. IgA was significantly elevated in patients with anaemia of mild to moderate severity and in patients with hookworm only (P < 0.05) while sIgA was significantly elevated in all subgroups compared to controls (P < 0.05). IgM was significantly elevated in-patients with marked anaemia, in-patients without anaemia and in those with hookworm infection associated with other parasites (P < 0.05). The difference in IgG levels between patients and controls was not significant (P 0.1). C4 was significantly elevated in patients with marked moderate anaemia and those with hookworm only (P < 0.05) while C3 levels were not significantly different in the subgroups compared with controls. These results suggest the possibility of polyclonal B-cell activation by T-independent antigens such as the polysaccharide cuticular antigens of the hookworms and the stimulation of the classical pathway of the complement system. PMID- 11379440 TI - The Idikan adult mortality study. AB - The Idikan adult mortality study is designed to explore the usefulness of the verbal autopsy methodology in the determination of cause-specific adult mortality. Such data have been largely unavailable in developing countries. Members of a stable urban community (4333 adults) were registered in their family units and followed up every 3 months to ascertain deaths, exits and new entries to the study population. Deaths were investigated by means of a verbal autopsy, which was administered by a trained interviewer to the relative closest to the decedent. Two physicians independently studied the results and assigned a cause of death, and these were then confirmed or resolved by a senior physician colleague. Hospitals where deaths occurred were visited and assignable cause of death was obtained where available and compared with the cause assigned by verbal autopsy. Only 8 of the 60 investigated deaths were confirmed to have occurred in a hospital in the first 2 years of the study. The cause of death assigned by verbal autopsy agreed moderately both between the independent coders as well as with the hospital assigned cause of death using the Kappa statistics for agreement beyond chance. We therefore concluded that verbal autopsies appear moderately useful for adult death statistics (Kappa = 0.23 to 0.1). Accumulation of more hospital deaths is needed, however, to make more definite conclusions about the validity of the technique. PMID- 11379441 TI - Anti-diarrhoeal and gastro-intestinal potentials of the aqueous extract of Phyllanthus amarus (Euphorbiaceae). AB - The anti-diarrhoeal and gastro-intestinal protective potentials of aqueous extract of leaves of Phyllanthus amarus were investigated in mice. Graded doses of the aqueous extract (100-800 mg/kg) administered orally produced a dose related inhibition of gut meal travel distance in normal mice. The highest intestinal transit inhibition of 31.65% was obtained with 400 mg/kg. In castor oil induced diarrhoea in mice, P. amarus extract (400 mg/kg) delayed the onset of diarrhoea, reduced frequency of defecation and reduced gut meal travel distance significantly resulting in intestinal transit inhibition of 79.94% compared to 86.92% produced by morphine (100 mg/kg). In addition, the activities of some intestinal mucosa enzymes (maltase, sucrase, lactase and alkaline phosphatase) in mice pretreated with extract before castor oil were not as severely depressed as those in the control (castor oil treated mice). Phytochemical screening revealed the presence of many secondary metabolites. The results are discussed with a view to establishing the basis of the use of this plant in traditional medicine for treatment of diarrhoea and other gastrointestinal disorders. PMID- 11379442 TI - Comparative analysis of blood plasma epidermal growth factor concentrations, hormonal profiles and semen parameters of fertile and infertile males. AB - The relationship between male reproductive function and the blood plasma level of epidermal growth factor (EGF) is of interest in the light of the role that circulating EGF appears to play in regulating mouse spermatogenesis. We measured the concentrations of EGF in the blood plasma of 39 fertile men (sperm count > 20 x 10(6)/ml) and compared them with those of 31 infertile men (sperm < 20 x 10(6)/ml). Blood plasma levels of follicle stimulating hormone (FSH), luiteinising hormone (LH), prolactin and testosterone were also determined. The infertile patients had mean blood plasma EGF concentrations of 0.75 +/- 0.10 ug/L. The value was significantly lower than that of the fertile group (1.28 +/- 0.14 ug/L; P < 0.005). There were statistically significant differences between the fertile and infertile groups in sperm count, sperm viability, mean forward progression, testosterone, LH and FSH (P values between 0.0001 and 0.023). There was no significant difference in the prolactin concentrations between the two groups. Although overall average blood plasma EGF concentrations are significantly lower in the infertile males, regression analysis failed to reveal any direct relationships among the various parameters studied. PMID- 11379443 TI - Modulation of carbon tetrachloride-induced lipid peroxidation and xenobiotic metabolizing enzymes in rats fed browned yam flour diet. AB - The modulatory effect of browned yam flour diet, a dietary, staple in south western Nigeria, on carbon tetrachloride (CCl4)-mediated lipid peroxidation and on the activities of liver microsomal and cytosolic enzymes was studied in male rats. Browned yam flour diet fed at the level of 25% and 50% to rats for 5 weeks significantly reduced the lipid peroxidation induced by CCl4 (0.5 ml/kg/wk) administered two weeks after starting the animals with the diets. The diets elicited 62% and 79% reductions in CCl4-mediated peroxidation, respectively, in the absence of exogenously added oxidants. The activities of microsomal aniline hydroxylase (AH), aminopyrine-N-demethylase (APD), pentoxyresorufin-O-dealkylase (PROD) and cytosolic GSH S-transferase (GST) were increased when rats were fed the 25% or 50% browned yam flour diets. Browned yam flour fed at the level of 25% to rats decreased the CCl4-mediated reduction in the activities of microsomal AH, APD, PROD and GST by 64%, 28%, 58% and 25%, respectively, and by 82%, 48%, 83% and 55% when rats were fed with 50% of the diet. The results suggest that browned yam flour diet could protect against chemically-mediated lipid peroxidation and tissue damage possibly by scavenging chemically generated reactive species and enhancing carcinogen-detoxifying system. PMID- 11379444 TI - Caudal anaesthesia in the clinical assessment of painful anal lesions. AB - The clinical evaluation of patients who present with painful anal conditions is often incomplete because of the need to avoid distress that digital examination engenders. Diagnosis is then based on the history and other findings on clinical examination. This is associated with a risk of missed and delayed diagnosis, delay in the initiation of appropriate therapy and the use of alternative investigation modalities which may not be necessary if full clinical evaluation had been done. In this communication, the experience with the use of caudal anaesthesia in the outpatient and ward setting to evaluate painful anal conditions is presented. The result shows that the technique is safe, and allows the immediate and complete evaluation of these conditions. The anaesthesia is short lasting and no morbidity was observed in this study. PMID- 11379445 TI - The effect of different dietary fats on gastrin levels in the pyloric antrum and plasma of weaner and adult Wistar rats. AB - The effect of dietary fats on gastrin in the pyloric antrum and plasma of Wistar rat was examined. Two different age groups of rats were fed 3 different diets in which fat was in the form of menhaden oil (MO), hydrogenated coconut oil (CO) and safflower oil (SO) respectively. Control groups were fed on normal laboratory diet. Each diet was isoenergetic and no group showed significant differences in either food intake or weight gain during the experiment. Weaner rats fed on MO diet exhibited significant reduction in both antral (P = 0.047) and plasma (P = 0.002) gastrin concentrations when compared with age-matched controls. Likewise, adult rats fed the MO diet exhibited significant reduction in both antral (P = 0.008) and plasma (P = 0.002) gastrin concentrations. In addition, adult rats fed the CO diet exhibited significant reduction in both antral (P = 0.047) and plasma gastrin (P = 0.002) concentrations. Rats from both age groups fed the SO diet exhibited no significant differences in gastrin concentration when compared with their respective control groups. These data indicate that the composition of dietary fat can have profound effects on both tissue and plasma concentrations of gastrin in rats. PMID- 11379446 TI - Seasonal variations of hydroxyanthraquinone content of cultivated Cassia spectabilis DC. AB - Seasonal variations and spectrophotometric estimation of the anthraquinone content of cultivated Cassia spectabilis DC are presented. Concentrations of anthraquinones peaked in the leaves (1.03%) at flowering Significant (P < 0.05) variations in anthraquinone content were observed in the cultivated species. Anthraquinones reached peak levels during the months of September to January (dry season). There was a significant (P < 0.05) drop in anthraquinone content during the rainy season. The diagnostic microscopic features of this little studied species are also described. This study has provided vital information on the best period of harvest and seasonal distribution of anthraquinones in the leaves of the species. PMID- 11379447 TI - The nutrient content and effectiveness of rice flour and maize flour based oral rehydration solutions. AB - The effectiveness of rice flour and maize flour based oral rehydration solution (ORS) in treating mild and moderate dehydration was studied among 88 children aged to 6 to 42 months at the Oni Memorial Children Hospital, Ibadan. In this randomized clinical trial, fifty children were treated with rice-based ORS while fifty-three received the maize-based ORS, over a four hour period. There was clinical improvement in the signs of dehydration with a significant increase in body weights and a significant fall in the packed cell volume following the administration of the two preparations. The maize-ORS contains 4.3 g protein, 179.3 Kilocalories and rice-ORS contains 3.6 g protein and 152.1 Kilocalories per litre of the preparation, respectively. It is concluded that rice and maize based ORS were acceptable and effective in oral rehydration of diarrhoea patients. They may also contribute to the nutrient intake of patients with (mild to moderate) 'some' dehydration. However other sources of energy, protein and mineral intake should be continued in these patients. PMID- 11379448 TI - An investigation of the frequency of co-existence of osteophytes and circumscribed full thickness articular surface defects in the knee joint. AB - Osteophytes are osteochondral repair tissues formed usually at the margins of synovial joints in response to a more central full thickness articular cartilage defect. These defects can be managed using the autologous chondrocytes implantation technique or with composite osteochondral grafting. Materials for osteochondral grafting of circumscribed full thickness articular surface defects may be obtained from osteophytes. In this study, the frequency of co-existence of these two intra-articular lesions was studied prospectively in patients undergoing therapeutic arthroscopy for painful knee conditions. Thirty-three of 88 knees (37.5%) had full thickness articular surface defects and 23 of these (69.7%) had associated osteophytes formation. PMID- 11379449 TI - A comparative study of students' performance in preclinical physiology assessed by short and long essays. AB - The performance of 540 medical (MBBS) and 55 dental (BDS) students in short essay questions (SEQs) in preclinical physiology was compared with their performance in long essay questions (LEQs). The cohort was made up of 88 repeating and 452 non repeating MBBS students and 12 repeating and 43 non-repeating BDS students. The SEQs and LEQs used for the study were those administered to the students in the paper II of the Part I MBBS/BDS examination. The results showed that all the students did significantly better in LEQs than in SEQs. When the students were sub-divided into repeaters and non-repeaters, performance in LEQs was still significantly better in all cases, except in the BDS repeaters where the difference was not significant. The study also showed that the BDS students did significantly better than the MBBS students in LEQs. Further analysis showed that the latter was due to a much better performance in LEQ 4 by the BDS students. In the SEQs, MBBS non-repeaters did better than repeaters while the situation was reversed with the BDS students. For all categories of students, on no occasion was the mean score in the LEQs or the SEQs up to 20.0, which is the 50% score and pass mark in this examination. The likely reasons for the better performance of the students in LEQs than in SEQs were discussed. The better performance of the BDS students in LEQs is believed to be due to the double exposure time of the BDS students to the topics from which LEQ 4 is drawn. From our results, it was concluded that LEQs are more useful than SEQs in assessing these students. In addition, the highly significant cross-correlation between scores in questions 1, 2, 3 and 4 suggested that the inherent problem of examiners' subjective judgement in essay marking could be minimized in both SEQs and LEQs. Finally, failure to achieve a mean score of 50% (pass mark) in both SEQs and LEQs suggested that the students are weak in essay writing. Possible reasons for this were suggested. PMID- 11379450 TI - Effects of adrenoceptor blockers on the glycemic response to nicotine in thyroidectomised rats. AB - The effect of adrenoceptor blockers on nicotine induced hyperglycemia were studied in fasted normal and thyroidectomised rats. Blood glucose was estimated using the modified glucose oxidase method. The control experiments consisted of sham operated rats with intact thyroid glands. Pre-treatment of the rats with either the alpha adrenoceptor blocker prazosin or the beta adrenoceptor blocker propranolol before intravenous injection of nicotine (50 micrograms/kg) significantly reduced the hyperglycemia induced by nicotine in normal rats, while a combination of propranolol and prazosin abolished nicotine induced hyperglycemia. In the thyroidectomised group, nicotine also caused hyperglycemia. However, the basal glucose level and peak of glycemic response were lowered compared to that in the control group. The results therefore seem to suggest that both alpha and beta adrenoceptors are involved in nicotine-induced hyperglycemia in the rat. PMID- 11379451 TI - A review of 100 cases of supracondylar fractures in children seen in Ibadan. AB - Supracondylar fracture of the humerus in children is one of the commonest fractures in children of school age all over the world. The experience of the mode of presentation, mechanism of injury and the different modalities of treatment in the University College Hospital, Ibadan is presented. The results suggest that severely displaced supracondylar fractures in this environment are better managed with open reduction and internal fixation. PMID- 11379452 TI - E-test method of antimicrobial susceptibility testing of Neisseria gonorrhoeae for routine diagnostic service. AB - The minimising inhibitory concentrations of four antimicrobial agents for 64 clinical isolates of Neisseria gonorrhoeae including 26 penicillinase producing strains (PPNG) as determined by E test, a recently developed method for sensitivity testing, were compared with those of agar dilution method using isosensitest (IST) agar. The medium was supplemented with either 5% lysed horse blood alone or with both lysed horse blood and 1% vitox defined supplement. The E test MICs compared closely with those obtained by agar dilution with essential agreements within +/- 1 log2 dilution being over 90% with all test antibiotics on medium that did not contain vitox, and between 71 and 93% on medium containing vitox. The Pearson's correlation coefficients ranged from 0.84 to 0.96 on either medium formulation. Excellent categorical agreement was obtained for all isolates with ceftriaxone, ciprofloxacin and tetracycline, while the E test gave a minor categorical discrepancy for two isolates with penicillin. We conclude that the E test is as reliable as conventional agar dilution method for MIC testing of N. gonorrhoeae in a routine laboratory. PMID- 11379453 TI - A case report of anorexia nervosa. AB - Although there are a few reports of Anorexia Nervosa in non-white populations some authors believe that the prevalence of anorexia Nervosa is as common in African populations as it is in Western countries. A case is presented of Anorexia Nervosa in a Nigerian female with features that strictly fulfill the ICD 10 diagnostic criteria. The difficulties encountered in the treatment of such disorders as well as the recurrent nature of the disorder were also highlighted in the patient's presentation. PMID- 11379454 TI - Fracture of the penis--diagnosis and management. AB - Fracture of the penis is an uncommon injury, but it occurs most commonly during over enthusiastic sexual intercourse. Diagnosis is not difficult but serious complications such as urethral rupture and corporo-urethral fistula may occur. Management should be by early surgical exploration, with evacuation of the haematoma and repair of the tear in the tunica albuginea in order to minimize the length of patient's stay in hospital and avoid complications such as penile deformity and painful penile erections. PMID- 11379455 TI - Primary hyperparathyroidism as a cause of multiple long bone fractures in a young Nigerian adult. AB - Primary hyperparathyroidism is a recognized though relatively rare cause of multiple limbs fractures. We report a case in a young Nigerian adult. It is emphasize that primary hyperparathyroidism may be asymptomatic and therefore under-recognized in this environment because it is very uncommon. PMID- 11379457 TI - Variability in the relationship between serum creatinine and creatinine clearance in hypertensives and normotensives with normal renal function. AB - Variabilities exist in the relationship between serum creatinine (Se-C) and creatinine clearance (CCr) due to the influences of age, muscle mass and gender on creatinine production. We studied this variability in a group of hypertensives (n = 62, 30 male, age 44 +/- 7 years) and normotensives (n = 90, 47 male, age 42 +/- 10 years) with normal renal function (Se-C < 1.5 mg/dL). There was a wide scatter of CCr about each Se-C value. Se-C was higher in males than females and in hypertensives than normotensives, but the differences were not statistically significant. Mean Se-C for the whole group was 0.91 +/- 0.23 mg/dL and corresponded to CCr of 93 +/- 23 mL/min and 93 +/- 21 mL/min before and after adjustment to a body surface area (BSA) of 1.73 m2. Adjusted CCr was significantly lower in hypertensives than normotensives in males (86 +/- 20 mL/min Vs 99 +/- 21 mL/min, P < 0.01) and females (84 +/- 19 mL/min Vs 96 +/- 19 mL/min, P < 0.01). Crude CCr was significantly higher in men than women (P < 0.05) in the whole group but not significantly in the normotensive or hypertensive subgroups. After adjustment, the differences diminished and became insignificant in the whole group. Some subgroups had identical mean Se-C values but different adjusted CCr. A particular Se-C value did not always indicate a particular CCr. Adjustment of CCr to 1.73 m2 BSA reduced the variability between Se-C and CCr and the difference between the genders. CCr showed significant difference in renal function between subgroups where Se-C did not. PMID- 11379456 TI - Response of laboratory staff to vaccination with an inactivated Rift Valley fever vaccine--TSI-GSD 200. AB - Laboratory staff and students were vaccinated with a formalin-inactivated rift valley fever (RVF) vaccine. This study showed that the vaccine used (TSI-GSD 200) was able to bring about the production of antibodies in recipients. For the production of a high titered antibody response, three doses of the vaccine were required. One or two doses of the vaccine did not produce a greater than four fold rise in antibody titre. A greater than four-fold rise in antibody titre following vaccination, is considered significant. The complete dose of the vaccine, that is, three doses, was necessary for protection. This study also showed that the haemagglutination inhibition (HI) test was capable of detecting antibodies, few weeks post vaccination. Though such HI antibodies broaden with time, it could be used for screening purposes and a more specific test, e.g., plaque reduction neutralisation (PRN) test used for confirmation of such results. PMID- 11379458 TI - Prostate specific antigen in the Nigerian African. AB - The biological characteristics of prostate-specific antigen were studied in the Nigerian African. Two hundred and fourteen persons were selected for the study. The group was made up of 59 apparently healthy men (age range 22-76 years), 58 men (age range 40-91 years) who had biopsy proven diagnosis of cancer of the prostate gland, 81 men (age range 46-87 years) who had biopsy proven benign prostatic hypertrophy (BPH) and 16 women (age range 23-47 years). Their median ages were 53 years, 66 years, 66 years and 27 years, respectively. The median PSA value among the control population was 0.7 ug/L (range 0.1-4.3 ug/L) and 98.3% of them had PSA of less than 4 ug/L. No person below 50 years of age had PSA value greater than 2 ug/L. There was a significant correlation between age and serum PSA value over the entire age range (r = 0.523; P = 0.001). In the female study group, 4(25%) had detectable values the cause of which could not be determined. The median PSA value among the BPH patients was 8.5 ug/L (range 0.2-350 ug/L). In this group 37% of them had values below 4 ug/L and 54.3% of them had values less than 10 ug/L. Among the patients who had cancer of the prostate gland the median PSA value was 92.6 ug/L and the mode was greater than 350 ug/L (32% of the patients); 10% and 20% of them had values below 4 ug/L and 10 ug/L respectively. It is concluded that the trend is the same as in the studies and there is the need for prostate gland volume studies and evaluation of women who have detectable PSA values. PMID- 11379459 TI - Electrical (faradic) stimulation versus active mobilization exercise in the physical management of post-surgical temporomandibular joint hypomobility. AB - This clinical study compared the efficacy of faradic stimulation and active mobilization exercises in the physical management of patients with post-surgical immobilization of the temporomandibular joint (TMJ) resulting in hypomobility of the joint. Eight volunteer dental patients with post-surgical immobilization TMJ hypomobility at the University College Hospital (U.C.H.), Ibadan, Nigeria participated in the study. Duration of TMJ immobilization was between 6 and 10 weeks (mean 7.13 +/- 1.55). Patients were alternately assigned to two groups as they became available. Patients in group A received mild infra-red radiation to the TMJ region and faradic stimulation to the muscles that move the joint while patients in group B had mild infra-red radiation and TMJ mobilization exercises. Treatment continued until pain relief and full range of the TMJ were attained. However after three treatment sessions, attendance became irregular because the patients were satisfied with their recovery. Pain perception was measured using the visual analogue scale. Interincisal opening was measured using a pair of mathematical set divider and a measuring ruler. The results showed that both faradic stimulation and exercises significantly improved the interincisal opening and pain perception although electrical stimulation improved mouth opening more significantly than active exercise. PMID- 11379460 TI - The haematological profile of Nigerians with chronic renal failure. AB - In an attempt to add to existing sparse literature on the haematological profile in chronic renal failure (CFR) in Nigeria, we have undertaken a comprehensive haematological sturdy of 39 patients (male 27, female 12) age range 11-56 yr., (mean 28.8 +/- 11.8) who had established pre-dialytic CFR. The mean haematocrit was 24.1 +/- 6.7% (range 12-40%). Severe anaemia was found in seven (18%), mild to moderate anaemia in 27 (69%) whilst five patients were not anaemic. Haematocrit correlated inversely with the degree of renal failure as assessed by serum creatinine (r = -0.35, P < 0.05). Red cell morphology was variable but the majority of patients showed a normocytic, normochromic blood film. The reticulocyte counts/indices were low. The mean total white cell count was generally within normal limits, ranging from (2 to 10.5 x 10(9)/l), with a mean of 5.3 +/- 2.1 x 10(9)/l and striking eosinophilia in 5 patients. Platelet count ranged between 82 and 350 x 10(9)/l (mean 156.5 +/- 65.7 x 10(9)/l) with only 3 patients having a relatively low count of < 100 x 10/l. Prolonged bleeding time (BT) > 9 minutes occurred in 13 (25.6%). There was no significant correlation between platelet count and bleeding time r = 0.21, P = 0.34. No significant correlation was observed between serum creatinine and bleeding time r = 0.09, P > 05. The bone marrow showed predominantly normocellular marrow but 7 patients had hypocellularity. Myeloid: Erythroid ratio ranged between 1:1 and 10:1, (mean 3:6:1) and correlated positively with serum creatinine values. (r = 0.37, P = 0.048). Bone marrow storage iron was absent in two and reduced in six patients. Severe anaemia is a common feature in Nigeria patients with CRF and it strongly associated with the severity of the renal failure. The low reticulocyte count and the tendency for erythroid hpoplasia to occur with increasing severity of renal failure would necessitate the use of erythropietin in our patients. The increased bleeding tendency in some of the patients calls for caution in surgical procedures in these patient. PMID- 11379461 TI - Genital ulcers disease among sexually transmitted disease clinic attendees in Ibadan, Nigeria. AB - Genital ulcer disease (GUD) is a risk factor in the transmission of human immuno deficiency virus (HIV). The goal of this study is to estimate proportion, identify risk factors, and improve prevention and control of GUD. This is a retrospective study of 211 cases of GUD seen between 1993 and 1997 in an urban public sexually transmitted disease (STD) clinic. Genital ulcers form 7.6% of all STDs seen. Overall, genital herpes was commonest (89 or 42.25%). It was the predominant infection (84 or 44.7%) in the males, while lymphogranuloma venereum (52 or 24.7%) was in females. The peak incidence in both sexes occurred in the 20 29 age group. Males out numbered females by a ratio of 8:1. Most of the patients were single 114 (68.3%) and most 70 or 33.3% were students. Risk markers identified were: casual sex (103 or 53.5%) and multiple sexual partners (77 or 36.5%). Both were significantly higher (P < 0.05) in single patients. Self treatment, use of multiple drugs and incomplete course of antibiotics were also common. The need to intensify STDS education programmes to all occupational groups and to students in particular is highlighted. Commercial sex workers require periodic education, screening and treatment. PMID- 11379462 TI - Distribution of ABO and RH genes in Nigeria. AB - Phenotype and gene frequencies of ABO and RH (D) systems were studied in 37,846 random blood donors in five zone of Nigeria (South West) (Yoruba)--Zone A, North West (Hausa-Fulani)--Zone B, Plateau (Birom)--Zone C, South East (Igbo)--Zone D and North East (Kanuri)--Zone E). We assessed the micro differences of genetic markers of ABO and RH blood groups between the ethnic groups in the ABO and RH blood group systems. Gene frequencies were ABO *O = 0.7068, ABO *A = 0.1490, ABO *B = 0.1443, RH *D = 0.8150 and results are similar to those earlier reported. Phenotype frequencies of the blood groups were in agreement with Hardy-Weinberg equilibrium expectations, except in two zones B and C where deviation was thought to be due to a high frequency of blood group AB. PMID- 11379463 TI - The predictive value of white cell count in assessing clinical severity of sickle cell anaemia in Afro-Caribbeans patients. AB - The relationship between each of haemoglobin concentration (Hb), red cell count (RCC), white cell count (WBC), platelet count (PLT), mean corpuscular volume (MCV), red cell width difference (RWD), and average number of acute admissions per year (AVEADM), were determined in a cross-sectional study of 128 Afro Caribbean sickle cell anaemia patients attending the Sickle Cell Disease Clinic of Central Middlesex Hospital in London for a mean of 7.6 patient-years. The diagnosis of sickle cell anaemia was made by both haematological and DNA analyses. The haematological parameters were determined using Coulter S automated counter during the steady state periods while the AVEADM was computed from all admissions for painful crisis, acute anaemia and acute chest syndrome, priapism, and acute stroke. Haemoglobin F level was determined by HPLC. Analysis was done in the paediatric and adult patients separately. There were no significant correlations between any of the parameters and AVEADM in the paediatric group. In adult patients, there was significant positive correlation (P < 0.05) between WBC and AVEADM. WBC has a negative correlation with Hb concentration and HbF level. WBC 10 x 10(9)/L and above is associated with Hb and HbF level below the mean for the group. WBC is lower but not significantly, in patients with single alpha-gene deletion than in those without deletion (P = 0.06). This study suggests that higher WBC count may suggest possible increased hospital admission, lower Hb concentration, and lower HbF level, in adult patients, and that, as a single parameter it can be of value in the assessment of patients with sickle cell anaemia. Possible mechanisms for these findings are also suggested. PMID- 11379464 TI - A comparison of topical honey and phenytoin in the treatment of chronic leg ulcers. AB - In view of the reports that phenytoin and honey are useful in the healing of wounds, a comparison of their topical use in the treatment of chronic leg ulcers was carried out. Fifty cases of chronic leg ulceration were studied, each for a period of four weeks. They were assigned into three groups for honey, phenytoin/honey mixture, and phenytoin topical treatment. Overall mean duration of the ulcers was 56.5 months while the mean(s.d.) size was 3339 (5193) mm2. Mean percent reduction in size in the group treated with honey, 27.0 (36.9), was not significantly different (H = 0.26; 2 df; p = 0.88) from that of the mixture group, which was 25.9 (46.4), and from that of the phenytoin group which was 35.5 (53.2). This percent reduction in size was significantly greater, (H = 7.69; 2 df; P = 0.02), during the first week in the phenytoin group than in the other groups. Four of the cases progressed to complete healing at the end of four weeks with phenytoin. Pain score difference (using a graduation scale from 0 to 10) at the end of the four week treatment, was, 1.8 (1.7) in honey group, 2.0 (1.3) in mixture group and 3.6 (2.4) in phenytoin group. This difference was not significant, (H = 3.09; 2 df; P = 0.21). Our study suggests that phenytoin may be superior to honey as a topical agent in the treatment of chronic ulcers. PMID- 11379465 TI - The selective use of duplex scanning in the pre-operative assessment of primary and uncomplicated varicose veins identifies patients with 'early' morphological disease. AB - The standard management of primary lower limb varicosities (stripping of the long saphenous vein (LSV) with avulsion of the varicosities) is based on the assumption that the disease originates from primary valvular insufficiency and incompetent perforating veins. It has recently been suggested that the LSV may be spared in patients with superficial varicosities without incompetence of this vein, i.e., early disease. However, pre-operative identification of suitable patients for conservative surgery has proved difficult. In this study, we employed selective duplex scanning in cases with primary varicosity in whom sapheno-femoral and sapheno-popliteal incompetence had been excluded both clinically and with the hand-held Doppler (52 limbs [10% of new referrals]). We found that the varicosities in these limbs were of 3 types: (a) those arising independent of superficial vein incompetence (Type 1); (b) those associated with segmental LSV incompetence (Type 2); (c) those in whom incompetence of the sapheno-femoral junction and LSV was missed (Type 3). We therefore suggest that long saphenous varicosity may progress from Type 1 through to Type 3 with each type representing an increase in severity of the disease. We suggest that selective use of duplex scanning will assist in identifying those patients with early stages of the disease who can then be treated effectively with LSV-sparing surgery. PMID- 11379466 TI - Personal hygiene as an alternative to griseofulvin in the treatment of tinea cruris. AB - Two hundred and twentyfive (225) consecutive patients comprising 193 males (85.7%) and 32 females (14.2) were randomized into two groups to evaluate the effectiveness of personal hygiene in the treatment of tinea cruris. This double blind trial, which lasted for 84 days, took place at the dermatology clinic, Jericho Hospital, Ibadan. The first group of 112 subjects (96 males, 16 females) received griseofulvin 500 mg daily for 10 days while the other group of 113 subjects (97 males, 16 females) received placebo and personal hygiene (restricted bathing). There was informed consent from each patient. Presence of hyphae in the affected groin was determined by microscopy and culture methods. Patients recorded the occurrence of pruritus daily in the health diary given to them. The result showed that hyphae had almost disappeared in the griseofulvin group by 4th week but resurfaced in 59.8%, 78.6%, 83.0%, 87.5% of the patients by the 8th, 10th, 11th, 12th week, respectively. In the placebo restricted bathing group, proportion with the positive hyphae was initially high in the second week (88.0%), third week (90.0%) but declined rapidly after the 4th week (60.0%) until nearly zero by the 10th week and onwards. It is therefore concluded that simple personal hygiene and health education without medication are more effective and cheaper than pharmaco therapy like griseofulvin in the treatment of tinea cruris. PMID- 11379467 TI - Plasma nicotinic acid level in chloroquine treated rats. AB - The effect of chloroquine phosphate on plasma nicotinic acid levels in adult male albino rats was investigated. Pyrogen free chloroquine phosphate in physiological saline was administered subcutaneously to rats in a dose of 15 mg/kg body weight daily for eleven succeeding days in the treatment group. A control group was given equal volume of physiological saline daily for eleven succeeding days. Nicotinic acid concentration in the plasma was determined [1] Plasma nicotinic acid level was found to be significantly reduced (P < 0.01) throughout the duration of treatment. No change was observed in the control group. The significant reduction of plasma nicotinic acid level observed in this study may not be unrelated to competitive inhibition of the enzyme tryptophan dioxygenase by chloroquine phosphate. PMID- 11379468 TI - Bronchial asthma: a risk factor for hypertension? AB - Several attempts have been made to improve primary prevention of essential hypertension and many of these have been directed at avoiding the well known risk factors. Both asthma and hypertension are spastic disorders of smooth muscle, also asthmatics and hypertensives have been found to be salt sensitive. There is a suspicion that the similarities between these two diseases may predispose the individuals with one disease to the other, as pulmonary hypertension has been described during exercise-induced bronchoconstriction. We therefore, studied the blood pressure pattern during and after acute severe asthma (ASA) along with the frequency of hypertension in stable asthmatic patients. Two groups of patients were studied. Group 1 consisted of 12 patients with ASA (2 males, 10 females) with a mean age of 30 +/- 9.9 years. The mean blood pressure during attack of ASA (147 +/- 16.9/100 +/- 8.2 mmHg) was higher than the mean BP (132 +/- 8.3/82 +/- 7 mmHg) 2 weeks after discharge from hospital without treatment in all patients (P < 0.05). Group 2 included 134 asthmatic subjects in stable state (54 males, 80 females) with a mean age of 45 +/- 15 years and a range of 15-90 years. The overall frequency of hypertension was 37% with a proportion of 39% in males and 35% in females. Hypertension was defined as systolic blood pressure of > or = 140 mmHg and or diastolic blood pressure of > or = 90 mmHg. There was no difference between the frequency of attack of ASA in hypertensives (5.7 +/- 5.6 per year) and nonhypertensives (5.5 +/- 3.8 per year), P < 0.05. We concluded that transient elevation of blood pressure may occur during ASA. The frequency of hypertension among asthmatics is quite high and concurrent family history of hypertension and frequency of attack of ASA did not seem to determine the status of blood pressure. Patients with asthma should have regular blood pressure check during follow-up visits. PMID- 11379469 TI - Modified short-course chemotherapy of pulmonary tuberculosis in Ibadan, Nigeria- a preliminary report. AB - Over a 3 year period 3rd of April 1995 and 6th of April 1998 a controlled clinical trial of the modified short-course chemotherapy (SSC) in newly diagnosed cases of pulmonary tuberculosis in Nigeria was carried out. Between The SCC used was the one adopted from World Health Organisation/International Union Against Tuberculosis and Lung Diseases for developing countries by the Nigerian National Tuberculosis and Leprosy Control Programme (NTLCP). The regimen used consisted of streptomycin (S), isoniazid (H), Rifampicin (R) and pyrazinamide (Z) in the initial or intensive phase of 2 months. Ethambutol (E) was sometimes substituted for streptomycin. The continuation phase was 6 months of thiacetazone, (T) and isoniazid (H), i.e., 2SHRZ/6TH or 2EHRZ/6TH. Sputum conversion was 90% at the second month of treatment and there was no bacteriological relapse after 18 months of follow-up. Side effects were few and consisted mainly of acne vulgaris which occurred in twenty (20.6%) of 97 patients during the continuation phase. It is concluded that the 8-month chemotherapy regimen adopted by NTLCP is efficacious in treatment of smearpositive pulmonary tuberculosis (PTB). PMID- 11379470 TI - Treatment of endometriosis with depot medroxyprogesterone acetate: a preliminary experience. AB - The efficacy of depot medroxyprogesterone acetate in the treatment of endometriosis was assessed in 19 patients with severe diseases. Assessment was based on changes in subjective symptoms and signs at 4 weekly interval during treatment and after 52 weeks follow up, and changes in visible deposits and adhesions at laparoscopy before and after treatment. There were significant reductions in mean total subjective and symptoms scores, mean total R-AFS adhesions and implants scores, and mean additive diameter of implant scores at the end of treatment and follow up. Treatment success occurred in 75% of the patients, majority (66%) of whom had complete resolution of deposits. Side effects encountered include menorrhagia, break through bleeding, excessive weight gain, myalgia, breast pain, acne and delay in return of menses. It was concluded that DMPA an effective, cheap and readily available medication which is worth using in patients who can not afford the expensive alternatives. PMID- 11379471 TI - Soft tissue injuries of the face: a 10 year review. AB - A review of 1,173 soft tissue injuries seen and managed at the University College Hospital, Ibadan over a 10 year period was made. The age range was from 4 months to 85 years with a dominant age group range of 21-30 years. The commonest type of soft tissue injury was laceration while the commonest aetiology was road traffic accidents followed by falls. Males were generally more involved than females and the commonest complication was wound dehiscence. PMID- 11379472 TI - Second ipsilateral ectopic gestation after total salpingectomy: a case report. AB - A rare case of a second ipsilateral ectopic gestation occurring in the stump of the cornua three years after total salpingectomy for ruptured ectopic pregnancy is presented. PMID- 11379473 TI - Antiphospholipid antibody syndrome: two case reports and review of literature. AB - Antiphospholipid antibody syndrome (APAs) is an acquired multisystemic disorder characterised by hyper coagulation. It manifest clinically with arterial and venous thrombosis. It is not a rare phenomenon, but there are paucity of reports of this disorder in our environment. We present two cases of APAs with the hope that it will stimulate the awareness of clinicians in the recognition of this disorder in our environment. PMID- 11379474 TI - Creatinine clearance estimation from serum creatinine values: evaluation and comparison of five prediction formulae in Nigerian patients. AB - Predictability of creatinine clearance (CrCl) from serum creatinine would reduce the cost of renal care and obviate the need for 24-hours urine collection. Correlations have been established between serum creatinine (Scr) and 24-hour urinary creatinine clearance with derivation of various formulae. We have tested the applicability of these formulae in 34 Nigerian patients (22 males, 12 females) aged 18 to 58 years, (mean age 34.97 +/- 11.20 years) with established chronic renal failure (CRF) mean Scr level 742.26 +/- 388.15 mol/L. 32 age and sex matched healthy adults with serum creatinine values below 120 umol/l., served as controls. Serum creatinine and 24 hour CrCl levels were determined on two consecutive occasions. Creatinine clearance values were also derived from Scr using each of the established prediction formulae: Cockcrof and Gault [1] Gates [2] Hull et al [3]; Jelliffe [4]; and Mawer et al [5]. A relationship was sought between measured CrCl and the predicted values (derived) using the stated formulae. Regression equation were generated and correlation coefficient r, coefficient of determination r2, F-ration, prediction error, all defining the nature and strength of relationship were determined. We observed that good and statistically significant correlations exist between measured CrCl values and those predicated from the formulae (r ranging from 0.908 to 0.968 and r20.82 to 0.93 P = 0.000) and that a linear relationship exists in all cases. Cockcroft and Gault formula gave the highest coefficient of determination r2 = 0.94. It is concluded that the existing formulae are adequate for determining CrCl from Scr and should be frequently used in the long term follow-up of patients with Chronic Renal Failure (CRF) in our setting. PMID- 11379475 TI - Case report on hereditary non-polyposis colon cancer (HNPCC) in Nigeria. AB - The role of genetic factors in the etiology of colorectal cancers (CRCs) has recently been elucidated with the discovery of the mismatch repair. These genes are responsible for less than 5% of all cases of CRCs in Caucasian series. In this pilot study, tumors from 5 randomly ascertained CRC patients were subjected to microsatellite analysis, and two were microsatellite unstable. Both of these two patients had germline mutations in MSH2. If this finding can be confirmed in a larger series of patients, it suggests that MMR genes play an important role in the etiology of CRCs in Africa. PMID- 11379476 TI - Women's health in Nigeria: past, present and prospects for the next millennium. The Ninth Paul Hendrickse Memorial Lecture. PMID- 11379477 TI - The rise of essentialism and the medicalization of sexuality. AB - During the last decade of the 20th century, biological explanations of social behavior have been steadily growing in popularity, slowly replacing previously dominant sociocultural explanations. This trend is clearly visible in the field of sex research. The first part of the paper provides an overview of the theoretical foundations of the two confronted paradigms, evolutionary essentialism and social constructionism. Although the current dominance of essentialism is the outcome of various factors, social and scientific, a major role in its rise to prominence was played by mass media demand for simplistic, pop-scientific statements. In the second part of the paper, three currently propulsive areas in sexology are presented as case studies: (a) etiology of homosexuality, (b) gender identity of intersex children, and (c) re conceptualization of sexual disorders. Two conclusions are offered: 1) the demise of the 'nurture monopoly' was necessary for the further development of sexology; and 2) the present trend toward a 'nature monopoly' represents a new and serious danger--as shown by the emerging medicalization of sexuality. PMID- 11379478 TI - Surgical treatment of infected knee contracture after war injury. AB - From June 1991 till March 1994, sixteen patients with extension knee contracture as a war injury complication were operatively treated at the Department of Orthopedics, Zagreb University School of Medicine. Infection was present in all patients. During the same period of time, 32 patients with extension knee contracture as a war injury complication and free from the signs of infection were also treated at the Department and served as a control group. The mean age at the time of wounding was 28.3 and 31.8 years in the study and control group, respectively. Operative procedures of extensive intra- and extra-articular adhesiolysis of the knee were performed in all patients. The mean duration of immobilization following injury infliction was 5.5 and 4.9 months in the study and control group, respectively. The mean preoperative knee motion amplitude ranged from 8 degrees in extension to 37 degrees in flexion in the study group, and from 5 degrees in extension to 38 degrees in flexion in the control group. Postoperatively, the mean knee motion amplitude ranged from 2.8 degrees in extension to 97 degrees in flexion in the study group, and from 1.9 degrees in extension to 100 degrees in flexion in the control group. Treatment results did not depend on either the presence of infection in the region of injury or on the duration of preoperative knee immobilization. The treatment of infection was most important for the success of operative treatment of knee contracture. A significant role was also played by adequate intra- and extra-articular adhesiolysis with appropriate intensive postoperative rehabilitation. PMID- 11379479 TI - Changes of plasma fibronectin levels in pregnancy induced hypertension. AB - A number of laboratory tests are available for evaluation of hypertension in pregnancy. These tests can be used to either predict and/or prognosticate preeclampsia and other hypertensive disorders of pregnancy. The aim of this study was to evaluate alterations in fibronectin homeostasis in normotensive pregnancy and in hypertensive disorders of pregnancy subclassified into chronic hypertension, preeclampsia superimposed on chronic hypertension, and pregnancy induced hypertension. A prospective, longitudinal study was conducted in 115 pregnant women aged 20-39 years, divided into four groups: normotensive (n = 40), chronic hypertension (n = 18), preeclampsia superimposed on chronic hypertension (n = 20), and pregnancy induced hypertension (n = 37). Plasma concentrations of fibronectin were measured by using single radial immunodiffusion assay (RIA) in the 8th, 18th, 23rd, 28th, 32nd and 36th week of gestation. Plasma fibronectin concentration showed no significant changes in normotensive pregnancy, but was significantly elevated in the third trimester in women destined to become preeclamptic or with preeclampsia in whom it reached a mean (+/- SD) of 0.40 +/- 0.09 g/L in the 36th week of gestation. In the groups with preeclampsia superimposed on chronic hypertension and with pregnancy induced hypertension, there was a significant difference between plasma fibronectin concentrations in 32nd (p < 0.01) and 36th (p < 0.001) week of gestation compared with either other levels in the respective group (in the 8th, 18th, 23rd and 28th week of gestation) or those recorded in other groups in the same period of pregnancy. These results suggested that the measurement of plasma fibronectin might be of diagnostic value in preeclampsia but could not be considered a useful predictor for preeclampsia. PMID- 11379480 TI - Malignant mesothelioma in central and Eastern Europe. AB - To obtain data on mesothelioma incidence in Central and Eastern Europe, a short questionnaire was sent to 83 researchers from 19 countries. The questions referred to the approximate number of mesotheliomas diagnosed per year in the country, degree of asbestos consumption, and percentage of lung carcinomas attributable to asbestos. Answers were received from 12 countries. For some major asbestos producers and/or consumers, such as Russia and Ukraine, mesothelioma data were unavailable or unreliable. In various countries of Central-Eastern Europe, the crude incidence of mesothelioma appeared to be lower than in Western countries. The reported annual numbers of mesotheliomas were 120 in Poland, 133 in Romania, and 78 in Hungary. Among the countries with a population of 5 million or less, the highest incidence was observed in Croatia (46 cases per year, peritoneal tumors not included). Data on the percentage of asbestos-related lung carcinomas are lacking. The knowledge about asbestos related cancer in Central and Eastern Europe remains fragmentary. Further investigations in this relevant area of public health should be encouraged. PMID- 11379481 TI - Hepatorenal syndrome: new perspectives in pathophysiology and management. AB - Hepatorenal syndrome (HRS) is a unique form of acute renal failure occurring in patients with advanced liver disease. Despite the severe derangement of renal function and ominous prognosis, minimal pathologic abnormalities of the kidneys are found at autopsy. The kidneys, if transplanted, are capable of normal function, which supports the concept that renal failure is functional and potentially reversible. The pathogenesis of HRS is not completely known, but it is probably the result of an extreme underfilling of the arterial circulation secondary to an arterial vasodilation located in splanchnic circulation. Besides the renal circulation, all other extrasplanchnic vascular beds also appear to be vasoconstricted. The diagnosis of HRS is currently based on several widely accepted diagnostic criteria aimed at excluding nonfunctional causes of renal failure. Recently initiated therapeutic approaches lend a note of optimism to the future management of HRS. These include liver transplantation as definitive treatment for patients with end-stage liver disease, and introduction of new vasoconstrictor drugs with the preferential effect on the splanchnic circulation. The development of HRS after spontaneous bacterial peritonitis may be effectively prevented by the administration of albumin together with antibiotic therapy. PMID- 11379482 TI - Hyperhomocysteinemia in end-stage renal failure. AB - The end-stage renal disease (ESRD) population experiences an excess morbidity and mortality due to arteriosclerotic cardiovascular disease (CVD) outcomes. Specifically, event rates for myocardial infarction and stroke are 5- to 10-fold in ESRD patients on maintenance dialysis than in the general population. Recently, there is controlled evidence that hyperhomocysteinemia occurs more commonly than any of the traditional CVD risk factors in ESRD patients. Prolonged exposure of endothelial cells to homocysteine impairs the production of nitric oxide and endothelium-dependent vasodilatation, they combine with low-density lipoprotein cholesterol to produce aggregates that are taken up by vascular macrophages in the arterial intima (foam cells), produce aggregatory effects on the platelets, and decrease endothelial antithrombotic activity due to changes in the thrombomodulin function. Current treatment regimens for ESRD hyperhomocysteinemia, which are based on the pharmacological doses of folic acid (5 to 15 mg/day), frequently result in suboptimal lowering of Hcy concentrations. Other potential therapeutic approaches (such as oral N-acetylcysteine at 1.2 g/day) merit controlled investigation. PMID- 11379483 TI - Respiratory function in female workers occupationally exposed to organic dusts in food processing industries. AB - Respiratory consequences of work in food processing industry were studied in 764 female workers exposed to organic dusts associated with the processing of green and roasted coffee, tea, spices, dried fruits, cocoa and flour. A group of 387 female workers not exposed to respiratory irritants served as controls for the prevalence of acute (during work shift) and chronic respiratory symptoms. A greater prevalence of all acute and chronic respiratory symptoms was consistently found among exposed workers than among control workers. The highest prevalence of chronic respiratory symptoms was recorded for chronic cough (40%), followed by acute symptoms of dry cough (58.7%). The difference in the prevalence of chronic respiratory symptoms between the exposed and control workers was in general significant (p < 0.01 or p < 0.05). Mean acute reductions of lung function over the work shift were recorded in all of the studied groups; the mean across-shift decrease as a percentage of preshift values was particularly marked in FEF25 ( 26.7%), FEF50 (-21.6%), followed by FEV1 (-9.9%) and FVC (-3.7%). The preshift (baseline) values of ventilatory capacity were decreased in comparison to the predicted ones, and were lowest for FEF50 and FEF25. This finding indicated an effect of organic dust on small airways. Our analysis suggested that both dust exposure and smoking history contributed independently to these respiratory findings. Disodium cromoglycate (DSCG) significantly diminished across-shift reductions for FEF50 and FEF25 in a subgroup of the examined workers. Our data suggested the female workers employed in food processing industry to be at risk of developing both acute and chronic respiratory symptoms as well as ventilatory capacity impairment as the result of occupational exposures. PMID- 11379484 TI - The first case of leptospirosis in the Zadar area. AB - In September 1999, a 56-year-old butcher was admitted to the General Hospital in Zadar because of fever, headache, severe pain in the calf muscles and thighs, conjunctivitis, rash, hepatomegaly and jaundice. The initial diagnosis was septic shock, and the patient was admitted to the internal medicine ward. Microscopic agglutination test showed a fourfold rise of antibodies to Leptospira sejroe in the three serum samples. These serologic findings and laboratory findings of leukocytosis, thrombocytopenia, increased serum aminotransferases, blood urea nitrogen and creatinine, proteinuria and leukocyturia indicated that Leptospira sejroe was the etiologic agent of the disease in the patient presented. PMID- 11379485 TI - The first fatal case of Mediterranean spotted fever in Croatia. AB - The first lethal case of Mediterranean spotted fever in Croatia is described. A previously healthy, 58-year-old man was admitted to the hospital with high fever and rash. Several days later, severe anemia, leukopenia and thrombocytopenia developed, and the patient died about eight weeks after the onset of disease. Bone marrow biopsy showed hypercellularity of the cortical and trabecular bone structures with morphologically normal cells of all three hematopoietic lineages that were reduced due to regular nodular infiltrates. Serologic findings indicated that Rickettsia conorii infection was the etiologic cause of the patient's death. PMID- 11379486 TI - Polymyalgia rheumatica--an underdiagnosed disease. AB - Characteristic features of polymyalgia rheumatica, a widely underdiagnosed disease, are described. The features of the disease are illustrated by the authors' own experience in the treated patients, and compared with literature data. According to the authors' experience, patients with polymyalgia rheumatica are mostly treated for inaccurate diagnoses such as cervicobrachial or lumbosacral syndrome, seronegative rheumatoid arthritis, unexplained febrile state, or precipitated erythrocyte sedimentation rate of unknown etiology. This results is delayed diagnosis or the accurate diagnosis is never reached at all. PMID- 11379487 TI - Ethical dues in biomedical publications. PMID- 11379489 TI - General practice research in Australia: a timely reality check. PMID- 11379490 TI - Media and young minds. PMID- 11379488 TI - "Should we treat leuchaemia in children?". PMID- 11379492 TI - XIII International AIDS Conference, Durban, 9-14 July, 2000. Nelson Mandela argues for urgent action against HIV in Africa. PMID- 11379493 TI - Crevasse fall in the Antarctic: a patient's perspective. AB - As a veterinarian working on an Adelie penguin research program, I was to spend six months on an island off the coast of Mawson Station in the Australian Antarctic Territory. During a field training exercise on my third day at Mawson, I fell into a crevasse with my four-wheel-drive quad bike and was crushed between the bike and the crevasse wall six metres below the rim. I had hypothermia and abdominal injuries, and underwent two emergency surgical procedures at Mawson Station. Sixteen days after the accident, I was evacuated by helicopter and ship. Here, I describe my experiences. PMID- 11379494 TI - Sydney hailstorms: the health role in the recovery process. AB - We highlight the involvement of community health, public health and mental health staff in the recovery period after a natural disaster (hailstorm and ensuing wind and rain over three weeks) which, despite causing relatively few deaths or severe injuries, affected 20,000 families and caused damage estimated at $1.5 billion. Many families were given information and advice, and 383 individuals were referred for health assessment and management by doctors and other healthcare workers over an eight-month recovery period. PMID- 11379495 TI - Malaria in the Australian Defence Force during and after participation in the International Force in East Timor (INTERFET). AB - Malaria in Australian Defence Force members has been far more common in East Timor than in other recent overseas deployments. By six months after all 5,500 members of the International Force in East Timor had returned to Australia, 267 malaria infections had been reported to the Army Malaria Institute. Only 64 of those affected had their first clinical episode during their 4-5 months in East Timor, and about two-thirds of these infections were caused by Plasmodium falciparum. The remaining 212 soldiers developed their first symptoms after returning to Australia, and all but two infections were caused by P. vivax. After treatment, 44 soldiers had relapses of their vivax infections; 11 had a second relapse and two had a third relapse. These findings raise several issues about prevention and management of malaria in the ADF. PMID- 11379496 TI - Medical assistance to civilians during peacekeeping operations: wielding the double-edged sword. AB - Peacekeeping operations have become the main operational activity of the armed forces of the developed world over the past 10 years--a trend which appears likely to continue. Peacekeepers often remain deployed long after the armed conflict has ceased to help reconstruct civilian infrastructure. It is often possible to use the excess capacity of medical support units deployed with military forces to provide help to the local population. While this is appropriate immediately after a conflict when civilian clinics are overwhelmed, in the more prolonged reconstruction phase the seemingly simple clinical imperative to treat as many patients as possible becomes more complex. PMID- 11379497 TI - Cracking the code: how will the Human Genome Project affect life as we know it? PMID- 11379498 TI - Milestones in the Human Genome Project: genesis to postgenome. AB - The Human Genome Project (HGP) will change medicine and medical research irrevocably. The obvious gains in genetic knowledge from the HGP, together with the advances which will flow into bioinformatics, biotechnology and the potential for novel therapeutic agents, will ensure that the financial investment in the HGP is repaid many times over. The HGP's costs in terms of ethical and social issues remain to be determined, but it is to be hoped that these will not detract from the scientific and medical achievements. How did such an endeavour start, and what path did it follow? PMID- 11379499 TI - Just how long can we live? AB - In June this year, United States President Clinton and British Prime Minister Blair jointly announced that the human genome had been sequenced. In another year or so, this information should be assembled into a much more useful form than that in which it now exists. The advances which will be made possible by the Human Genome Project and new genetic technologies may well extend the human life span still further. PMID- 11379500 TI - The Human Genome Project: how do we protect Australians? AB - It is the moon landing of the nineties: the ambitious Human Genome Project- identifying the up to 100,000 genes that make up human DNA and the sequences of the three billion base-pairs that comprise the human genome. However, unlike the moon landing, the effects of the genome project will have a fundamental impact on the way we see ourselves and each other. PMID- 11379501 TI - The Human Genome Project in the dock. AB - From a scientific viewpoint, the Human Genome Project is actually not in the dock, nor even under reasonable suspicion of wrongdoing. Overwhelmingly, it will prove of benefit to humanity. However, from legal, ethical and other societal points of view, there are many problems already being considered by bioethicists, philosophers, religious experts, lawyers and others in dialogue with scientists. PMID- 11379502 TI - Social consequences of manufactured longevity. AB - The signs are that advances in biomedical sciences will add more years of "manufactured time" to life expectancy in industrialised countries, resulting in unprecedented rates of survival into older ages. Increasing longevity will force economic and social changes and the 20th-century revolution in social roles looks set to continue into the 21st century. PMID- 11379503 TI - What a louse! PMID- 11379504 TI - Shotgun suicide with a difference. PMID- 11379505 TI - Homicide or suicide? A jigsaw puzzle of incinerated human remains. PMID- 11379506 TI - General practice research in Australia, 1980-1999. AB - There has been a nearly fivefold increase in the amount of Australian general practice research published in 1990-1999 compared with the previous decade. The university departments of general practice and other university departments have been responsible for most of the research. GPs were involved in at least 60% of all of the research reviewed. Half of the research was found to be clinically pertinent to the front-line GP. The National Health Priority areas, introduced in 1994, were poorly represented, but it is probably too soon for this research to be published. There has also been little research on rural general practice. This review provides a starting point for classifying general practice and primary healthcare research in the future. PMID- 11379507 TI - Exploring the unknown: the challenges of a career in biomedical research. PMID- 11379508 TI - John Kerr and apoptosis. AB - On 14 March 2000, John Foxton Ross Kerr, Emeritus Professor of Pathology at the University of Queensland, received the Paul Ehrlich and Ludwig Darmstaedter Prize for his description of apoptosis, a form of cell death. The prize, which he shared with Boston biologist Robert Horvitz, is considered to be one of the most prestigious European awards in science, second only to the Nobel Prize. PMID- 11379509 TI - A revolution in rural and remote Australia: bringing health education to the bush. PMID- 11379510 TI - From Karrawingi the emu to Care factor zero. Mental health issues in contemporary Australian adolescent literature. AB - OBJECTIVE: To examine the depiction of mental health issues in contemporary Australian adolescent literature. This material might influence teenagers' understanding of mental illness and their willingness to seek help, and may provide insights into societal attitudes towards adolescents and the psychiatric problems encountered in this age group. DESIGN: Systematic examination of a sample of 94 books, the "Notable Texts" in the "Older Child" category of the Children's Book Council of Australia Awards for the years 1996, 1997 and 1998. MAIN OUTCOME MEASURES: Number of works referring to psychiatry; characters who experienced trauma, loss, psychiatric symptoms, qualified for a psychiatric diagnosis, exhibited suicidal behaviour or sought help; and the nature of treatments given. RESULTS: There was reference to psychiatry in over two-thirds of the works. In these books the image of psychiatry was generally negative or mixed. Most adolescent characters suffered major losses. Two-fifths of characters met criteria for a psychiatric diagnosis. The outcome for characters who satisfied criteria for a psychiatric diagnosis was more negative than those without. Of the 10 characters in the sample who committed suicide, eight had a psychiatric disorder. CONCLUSIONS: Psychiatric themes are a major component of the Australian teenage novel. Young people in contemporary Australian teenage literature are beset with trauma, loss and psychiatric disorders. Even when help is obtained, the outcome is seldom positive. This nihilistic view may increase hopelessness and pose a barrier for teenagers seeking help. PMID- 11379511 TI - Healthcare on the Internet: buyers beware. PMID- 11379512 TI - Medicine, the media and monetary interests: the need for transparency and professionalism. AB - Emerging evidence suggests that media coverage of medicine is increasingly promotional in nature. Recent Australian examples include misleading newspaper articles on an experimental cancer vaccine and a high profile television current affairs segment on a new influenza drug, which failed to disclose the industry ties of a key expert featured in the report. There are widening concerns that this problem in medical journalism may be exacerbated by the growing commercialisation of medical and scientific research, and the increasing ties between researchers, doctors and pharmaceutical or biotechnology companies. Closer links between industry and medicine are being explicitly encouraged both in academia and the health care sector for the mutual benefits they bring. However, these partnerships are the cause of growing unease within medicine. In the United States, rigorous legislation governing research protocols is being proposed, and in Australia new ethical guidelines covering industry-profession relationships are being promulgated. If one of the media's roles is informing the community about the business of health and medicine in a fair and accurate way, a cultural change in medical journalism is required. PMID- 11379513 TI - Life after medicine. PMID- 11379514 TI - Death and the comedian. PMID- 11379515 TI - A clinician with a passion for pathology. PMID- 11379516 TI - Service in three careers. PMID- 11379518 TI - A swimming program for children with asthma. Does it improve their quality of life? PMID- 11379519 TI - Pressure immobilisation bandages in first-aid treatment of jellyfish envenomation: current recommendations reconsidered. AB - OBJECTIVE: To evaluate whether applying pressure equivalent to that of pressure immobilisation bandages (PIB) causes release of additional venom from discharged jellyfish nematocysts. DESIGN: In-vitro experiment--the venom beads released from electrically activated Chiropsalmus sp. nematocysts were viewed under direct microscopy before and after applying 40 mmHg pressure (replicating the pressure of PIB); and saline washings of discharged nematocysts before and after applying pressure were tested for toxicity (time to ventricular standstill after injecting into live prawns). RESULTS: Applying 40 mmHg pressure caused the venom beads to visibly increase in size, consistent with pressure expressing further venom from the discharged nematocysts. First washings of the nematocyst shafts before compression produced ventricular standstill in prawns within 60 seconds (n=3); second washings did not produce standstill during 540 seconds of observation (n=3); and washings after applying 40 mmHg pressure showed a return of toxicity, with ventricular standstill in all prawns within 180 seconds (n=3). CONCLUSION: Discharged nematocysts are by no means empty and harmless. Applying pressure results in further release of nematocyst venom. The currently recommended practice of applying PIB in the initial treatment of patients stung by a jellyfish may exacerbate the envenomation, and thus should not be recommended. PMID- 11379520 TI - A possible case of intestinal myiasis due to Eristalis tenax. PMID- 11379521 TI - Envenomation by the billygoat plum stinging caterpillar (Thosea penthima). AB - We report the first case of envenomation by the billygoat plum stinging caterpillar, Thosea penthima Turner (Limacodidae). The sting, on the forearm, caused immediate burning pain and local wheal formation. Pain radiated up the arm and there was severe "crushing" chest pain lasting four hours. The local pain persisted for 10 hours and required opiate analgesia. PMID- 11379522 TI - Chromomycosis. PMID- 11379523 TI - Heartworm extraction. PMID- 11379527 TI - Would the pharmaceutical companies please mind their Ps and Qs, and their Xs, Ys and Zs. PMID- 11379528 TI - Australian general practice: the need for leaders and vision. PMID- 11379529 TI - R&D in general practice: time to move forward. PMID- 11379530 TI - The current situation of general practice. PMID- 11379531 TI - Setting the (medical) record straight. PMID- 11379532 TI - Incongruous infants: facial features in old paintings. PMID- 11379533 TI - The cough that packed a punch: is boxing a risk for tuberculosis transmission? PMID- 11379535 TI - [Ethics proposed by the Nippon Medical School for the 21st century: discussion]. PMID- 11379534 TI - Ulcerative "nintendinitis": a new kind of repetitive strain injury. PMID- 11379536 TI - Physician-assisted suicide and euthanasia: rebuttals of rebuttals--the moral prohibition remains. PMID- 11379537 TI - Statement on the application of the ToBI test for batch potency testing of tetanus vaccines for human use. PMID- 11379538 TI - Statement on the application of the CORROSITEX assay for skin corrosivity testing. PMID- 11379539 TI - Cytomegalovirus in "immunocompetent", critically ill, intensive care patients. PMID- 11379540 TI - Bacterial sepsis in patients with acquired immunodeficiency syndrome. PMID- 11379541 TI - Do gene variants really explain the heterogeneous outcomes in sepsis? PMID- 11379542 TI - Role of selectins in acute pancreatitis. PMID- 11379543 TI - A new frontier: posttraumatic stress and its prevention, diagnosis, and treatment. PMID- 11379544 TI - Sepsis-induced adrenal deficiency syndrome. PMID- 11379545 TI - A potential new therapy for sepsis: learning from experience. PMID- 11379546 TI - Specialty neurointensive care--Is it just a name or a way of life? PMID- 11379547 TI - Potential pitfalls in the practice of partial liquid ventilation. PMID- 11379548 TI - End-of-life care for children: bridging the gaps. PMID- 11379549 TI - A simple and cheap nostril retainer: silicone urine catheter. PMID- 11379551 TI - Bipolar cutting of muscle flaps. PMID- 11379550 TI - Is it true that the temporoparietal fascia has two layered structures? PMID- 11379552 TI - Tissue glue. PMID- 11379553 TI - A hinged, double-hook retractor design. PMID- 11379554 TI - Unusual intraoperative venous complication in a free DIEP flap. PMID- 11379555 TI - A simple method of identifying small postpalatoplasty fistulae. PMID- 11379556 TI - Comparative study of flap versus central segment expansion for the treatment of knee contracture: a preliminary study. PMID- 11379557 TI - Clarification on recertification in plastic surgery. PMID- 11379558 TI - A complication with the aesthetic use of Botox: herniation of the orbital fat. PMID- 11379559 TI - A simple aid to skin closure. PMID- 11379560 TI - Smoking and cool face lifts; is there a Buerger's type disease of the face? PMID- 11379561 TI - Cleft surgery in developing nations. PMID- 11379562 TI - Nonsurgical breast enlargement. PMID- 11379563 TI - Bibliography. Current world literature. Hematopoietic growth factors. PMID- 11379564 TI - Prostate cancer screening. PMID- 11379565 TI - [May a physician release information regarding a deceased patient?]. PMID- 11379566 TI - [Model for the medical director 2000; guidelines for a responsible, administrative and progressive role of the head of physicians in hospital administration]. PMID- 11379567 TI - [Is there a chance for the internist in the future?]. PMID- 11379568 TI - Diagnostic dilemma: papillary thyroid cancer with psammomatous calcifications. PMID- 11379569 TI - Models of capitated payment for endstage renal disease. PMID- 11379570 TI - Hyperthyroidism and criminal behavior. PMID- 11379571 TI - Cigarette lighter thumb. PMID- 11379573 TI - Molecular biology: a timely tool for further unraveling the "diabetes of stress". PMID- 11379575 TI - Retirement in the medical group practice setting. PMID- 11379574 TI - Will this magic bullet fly true? PMID- 11379576 TI - Being green: on the relationships between people and plants. PMID- 11379577 TI - In search of the silver bullet. PMID- 11379578 TI - Routine audit is an ethical requirement of screening. PMID- 11379579 TI - Effect of receiving a heart transplant. Surely it is too late for a randomised controlled trial. PMID- 11379580 TI - Effect of receiving a heart transplant. Research target should be stratification procedures and mechanisms of death. PMID- 11379581 TI - Effect of receiving a heart transplant. Peak exercise oxygen consumption is important predictor of outcome. PMID- 11379582 TI - Debate about blood pressure and epistaxis will continue. PMID- 11379583 TI - Principal variable is not what it seems in league tables. PMID- 11379584 TI - Reduction of postoperative mortality and morbidity. Research into modern anaesthesia techniques and perioperative medicine is needed. PMID- 11379585 TI - Reduction of postoperative mortality and morbidity. Little information was given on inclusion criteria. PMID- 11379586 TI - Dual blockade of renin-angiotensin system. Data do not support claimed benefit of combination over single treatment. PMID- 11379587 TI - Air travel and risk of venous thromboembolism. Passengers should reduce consumption of alcohol on flights. PMID- 11379588 TI - Air travel and risk of venous thromboembolism. Pulmonary embolism after air travel may occur by chance alone. PMID- 11379589 TI - Homoeopathy in malaria: head of infectious diseases replies. PMID- 11379590 TI - Sifting the evidence. Likelihood ratios are alternatives to P values. PMID- 11379591 TI - Sifting the evidence. Statistics must not be confused with science. PMID- 11379592 TI - Sifting the evidence. Perfect understanding seldom happens. PMID- 11379593 TI - Early elevation of soluble CD14 may help identify trauma patients at high risk for infection. AB - BACKGROUND: Elevated levels of soluble CD14 (sCD14) have been implicated in both gram-positive and gram-negative sepsis, and it has been associated with high mortality in trauma patients who become infected. METHODS: Eleven healthy volunteers and 25 adult trauma patients with multiple injuries and a mean Injury Severity Score of 32 participated. Whole blood was obtained at intervals. Immunohistochemistry was used to quantify membrane CD14 (mCD14), by flow cytometry and plasma levels of sCD14 by enzyme-linked immunosorbent assay. Analysis of variance and Student's T test with Mann-Whitney posttest were used to determine significance at p < 0.05. RESULTS: On posttrauma day 1, sCD14 was significantly different in the plasma of infected patients compared with normal controls (7.16 +/- 1.87 microg/mL vs. 4.4 +/- 0.92 microg/mL, p < 0.01), but not significantly different from noninfected patients. The percentage of monocytes expressing mCD14 in trauma patients did not differentiate them from normal controls; however, mCD14 receptor density did demonstrate significance in septic trauma patients (n = 15) versus normal controls on posttrauma day 3 (p = 0.0065). CONCLUSION: On the basis of our data, mCD14 did not differentiate infected and noninfected trauma patients, although trauma in general reduced mCD14 and elevated sCD14. Interestingly, 100% of patients who exceeded plasma levels of 8 microg/mL of sCD14 on day 1 after injury developed infections. Therefore, early high expressers of sCD14 may be at higher risk for infectious complications after trauma. PMID- 11379594 TI - Systemic inflammatory response syndrome score at admission independently predicts infection in blunt trauma patients. AB - BACKGROUND: Systemic inflammatory response syndrome (SIRS) score has been demonstrated to be an accurate predictor of outcome in critical surgical illness. To our knowledge, there is a paucity of data using SIRS score as a tool to predict posttraumatic infection. Our goal was to determine whether the severity of SIRS score at admission is an accurate predictor of infection in trauma patients. METHODS: Prospective data were collected on 4,887 blunt trauma patients admitted to a primary adult resource center designated trauma center over an 18 month period. Patients were stratified by age and Injury Severity Score (ISS). SIRS score was calculated at admission. SIRS was defined as an SIRS score > or = 2. Each patient was screened for infection by an infectious disease specialist. Those at high risk for infection were then monitored daily throughout their hospitalization. Centers for Disease Control and Prevention guidelines were used to diagnose infection. RESULTS: Of the 4,887 patients, 1,850 (38%) were admitted > 24 hours and evaluated for subsequent infection (mean ISS, 16 +/- 9; mean age, 43 +/- 19, SD). Thirty-one percent (577) of the patients acquired an infection. The mean hospital length of stay (20.2 days vs. 6.5 days) and mortality (7.8% vs. 2.7%) were significantly greater in the infected group (p < 0.001). Of the four SIRS variables (temperature, heart rate, white blood cell count, and respiratory rate), hypothermia and leukocytosis were the most significant predictors of infection (p < 0.001) when adjusted for age and ISS. SIRS scores of > or = 2 were increasingly predictive of infection when analyzed by multiple logistic regression analysis. CONCLUSION: An admission SIRS score of > or = 2 is a significant independent predictor of infection and outcome in blunt trauma. Daily SIRS scores may be a meaningful method of assessing postinjury risk of infection, and may initiate earlier diagnostic intervention for determination of infection. PMID- 11379595 TI - Practice management guidelines for the optimal timing of long-bone fracture stabilization in polytrauma patients: the EAST Practice Management Guidelines Work Group. PMID- 11379596 TI - Efficiency of cefepime in postoperative meningitis attributable to Enterobacter aerogenes. PMID- 11379597 TI - Fibrodysplasia ossificans progressiva. AB - Fibrodysplasia ossificans progressiva (FOP) is an extremely rare and disabling genetic disorder of connective tissue. The condition is characterized by congenital malformation of the great toes and by progressive heterotopic ossification of the tendons, ligaments, fasciae, and striated muscles. Fibrodysplasia ossificans progressiva occurs sporadically and is transmitted as a dominant trait with variable expression and complete penetrance. Reproductive fitness is low. There are fewer than 150 known patients with the disorder in the United States. A point prevalence of one affected patient in every 2 million of population has been observed. There is no sexual, racial, or ethnic predilection. The disease presents in early life; its course is unavoidably progressive. Most patients are confined to a wheelchair by the third decade of life and often succumb to pulmonary complications in the 5th/6th decade of life. At present there is no effective prevention or treatment. The recent discovery of overproduction of bone morphogenetic protein-4 in lesional cells and lymphocytic cells of affected patients provides a clue to both the underlying pathophysiology and potential therapy. The FOP gene has recently been mapped to human chromosome 4q 27-31. PMID- 11379598 TI - Testis-sparing surgery for benign testicular tumors in children. AB - PURPOSE: In this retrospective survey we identify preoperative and intraoperative criteria of nonmalignancy and analyze the result of conservative treatment of a testicular mass. MATERIALS AND METHODS: A total of 22 surgeons responded to a questionnaire concerning lesions treated during the last 15 years in children between 0 and 15 years old. By definition alpha fetoprotein and beta human chorionic gonadotropin are within the normal limits at this age. The results were evaluated regarding evolution, growth of the preserved testis and local or distant recurrences. RESULTS: Benign tumor of the testis (83 cases) represented 48% of all cases. Orchiectomy was performed in 27 cases and conservative treatment in 56. The final histopathological diagnosis was benign germinal tumor in 48 cases, cysts in 18, gonadal stromal tumor in 13 and rare lesions (lipoma, hemangioma) in 4. No definitive clinical criteria of nonmalignancy were identified but some symptoms were suggestive of nonmalignancy. Ultrasound results were more conclusive and provided the diagnosis of teratoma, epidermoid cyst and particularly simple cyst. The conservative treatment performed 56 times was a simple biopsy in 2 children with bilateral lesions and enucleation in 52. Enucleation was performed in 43% of cases using a pedicle clamp and in 50% with frozen section. The frozen section was changed in 12 cases due to therapeutic decision for preservation (10) and orchiectomy (2). There were no contradictions between the definitive histopathological examination and frozen section. Secondary orchiectomy was performed for neonatal granular tumor. Average followup in 56 cases of conservative treatment was 4.8 years (range 6 months to 15 years). Neither secondary testicular atrophy nor any local or distant recurrence was recorded. CONCLUSIONS: A testicular tumor in children has a 50% chance of being benign. Treatment selection according to some clinical, biological, radiological and frozen section findings should allow us to decide on testis-sparing surgery without additional oncological risk, and with an aesthetic, psychological and functional benefit. PMID- 11379599 TI - [Multislice computed tomography in clinical practice]. AB - This paper presents our first experience with multislice computed tomography emphasizing our technical protocols. Between November 1999 and October 2000, 8086 multislice computed tomographic examinations were performed. The choice in collimation, pitch, slice width and increment, multiplanar reconstructions and contrast enhancement were adapted to provide high resolution in the Z axis, high speed acquisitions and low exposure to the patient. Filming and archiving parameters are also detailed. Multislice CT provides excellent 3D imaging with high speed acquisitions and lower doses than previously reported with single slice CT. PMID- 11379600 TI - QT dispersion after exercise as an indicator of coronary stenosis. PMID- 11379601 TI - [Theoretical simulation of UV absorption spectra of carbazole and some homocyclic analogs]. AB - The UV absorption spectra structures of carbazole, fluorene and dibenzofurane are studied using the atom monopole-dipole interaction (AMDI) model and atomic dipolar polarisabilities and effective charge given by Fraga. The absorption spectra corresponding to these molecules and their self-associations are presented. The original obtained results are discussed and compared to previous works. PMID- 11379602 TI - Bond strength of binary titanium alloys to porcelain. AB - The purpose of this study was to investigate the bond strength between porcelain and experimental cast titanium alloys. Eleven binary titanium alloys were examined: Ti-Cr (15, 20, 25 wt%), Ti-Pd (15, 20, 25 wt%), Ti-Ag (10, 15, 20 wt%), and Ti-Cu (5, 10 wt%). As controls, the bond strengths for commercially pure titanium (KS-50, Kobelco, Japan) and a high noble gold alloy (KIK, Ishifuku, Japan) were also examined. Castings were made using a centrifugal casting unit (Ticast Super R, Selec Co., Japan). Commercial porcelain for titanium (TITAN, Noritake, Japan) was applied to cast specimens. The bond strengths were evaluated using a three-point bend test according to ISO 9693. Since the elastic modulus value is needed to evaluate the bond strength, the modulus was measured for each alloy using a three-point bend test. Results were analyzed using one-way ANOVA/S N-K test (alpha = 0.05). Although the elastic moduli of the Ti-Pd alloys were significantly lower than those of other alloys (p = 0.0001), there was a significant difference in bond strength only between the Ti-25Pd and Ti-15Ag alloys (p = 0.009). The strengths determined for all the experimental alloys ranged from 29.4 to 37.2MPa, which are above the minimum value required by the ISO specification (25 MPa). PMID- 11379603 TI - Manometric characteristics of the upper esophageal sphincter recorded with a microsleeve. AB - OBJECTIVES: We compared manometric recordings of the upper esophageal sphincter (UES) recorded with a miniature sleeve to those obtained using standard manometry. METHODS: The UES pressure of eight volunteer subjects was measured by station pull-through (SPT), by rapid pull-through (RPT), and with a microsleeve sensor for 30 min, followed by 15 min of esophageal acid infusion. Deglutitive UES relaxation recorded with a microsleeve and solid state sensor were compared. RESULTS: The UES pressure recorded with the microsleeve (25+/-9 mm Hg) was significantly less than that by SPT (114+/-18 mm Hg) or RPT (152+/-19 mm Hg), and was unaffected by acid infusion. Periods of low UES pressure were observed during long interswallow intervals (11+/-4, range 6-18 mm Hg). Deglutitive relaxation duration and intrabolus pressure measured with the microsleeve were less than those recorded by the solid state transducer. CONCLUSIONS: "Normal" UES pressure is heavily dependent on measurement technique; pressures obtained with a miniature sleeve are a fraction of those obtained by SPT or RPT. During periods of relative comfort with minimal swallowing, UES tone is approximately 10 mm Hg, similar to that during sleep. Volume modulation of deglutitive UES relaxation is demonstrable with a microsleeve, albeit with less precision than with a solid state transducer. PMID- 11379604 TI - Staying in touch. Working Committee for Media and Public Relations of the DGCZ. PMID- 11379605 TI - Adjuvant interferon alpha 2b in high risk melanoma - the Scottish study. AB - In 1989, the Scottish melanoma group initiated a randomized trial, comparing observation alone with 6 months' therapy with low dose interferon (given subcutaneously 3 MU day-1, twice weekly), for patients with primary melanomas of at least 3 mm Breslow thickness, or with evidence of regional node involvement. The trial was closed in 1993 with only 95 eligible patients randomized. There were no toxic deaths, and no patient failed to complete the treatment for reasons of toxicity. 6 months' treatment with low-dose interferon- resulted in a statistically significant improved disease-free survival for up to 24 months after randomization (P< 0.05). However, at a median follow-up of over 6 years, although there was an apparent improvement in disease-free survival (from 9 to 22 months), and overall survival (from 27 to 39 months), consistent with larger studies powered to detect such differences, these differences were not statistically significant. The data therefore suggest that 6 months of low-dose interferon is active, and confirm the importance of the large randomized studies, such as the UKCCCR AIM-High and EORTC trials, that seek to confirm a possible survival advantage for low or intermediate dose interferon. PMID- 11379607 TI - U.S. energy policy. Administration's energy plan is short on scientific details. PMID- 11379606 TI - The science of elections. PMID- 11379609 TI - U.S. science education. Lawmakers vie to shape NSF program. PMID- 11379610 TI - European higher education. Thirty nations pledge to harmonize degrees. PMID- 11379608 TI - Embryonic stem cells. Court asked to declare NIH guidelines legal. PMID- 11379611 TI - Human experimentation. Bioethics panel urges broader oversight. PMID- 11379612 TI - Cell biology. Protein clumps hijack cell's clearance system. PMID- 11379613 TI - Neurobiology. New clue to the cause of Alzheimer's. PMID- 11379614 TI - Solid state physics. Rubbery liquid crystal gives lasers a stretch. PMID- 11379615 TI - Gene silencing. A faster way to shut down genes. PMID- 11379616 TI - Quantum physics. Microscale weirdness expands its turf. PMID- 11379617 TI - Astronomy. 'Invisible' astronomers give their all to the Sloan. PMID- 11379618 TI - Academic collections. Japan shakes the dust off treasure trove of specimens. PMID- 11379619 TI - Egyptian science. A biotech gambit in the desert. PMID- 11379620 TI - Geology. Thermal features bubble in Yellowstone Lake. PMID- 11379621 TI - Evolution. Putting limits on the diversity of life. PMID- 11379622 TI - Hypoxia, fertilizer, and the Gulf of Mexico. PMID- 11379623 TI - Hypoxia, fertilizer, and the Gulf of Mexico. PMID- 11379625 TI - A global strategy to defeat invasive species. PMID- 11379624 TI - Hypoxia, fertilizer, and the Gulf of Mexico. PMID- 11379626 TI - The identity of plant glutamate receptors. PMID- 11379627 TI - Education. Workforce alternatives to graduate students? PMID- 11379628 TI - Ecology. Coral reef biodiversity--habitat size matters. PMID- 11379629 TI - Geophysics. Silent slip on the Cascadia subduction interface. PMID- 11379630 TI - Paleontology. Digging up fresh clues about the origin of mammals. PMID- 11379631 TI - Population ecology. Not all sheep are equal. PMID- 11379632 TI - Astrophysics. Mass outflow in active galactic nuclei. PMID- 11379633 TI - Mechanism of actin-based motility. AB - Spatially controlled polymerization of actin is at the origin of cell motility and is responsible for the formation of cellular protrusions like lamellipodia. The pathogens Listeria monocytogenes and Shigella flexneri, which undergo actin based propulsion, are acknowledged models of the leading edge of lamellipodia. Actin-based motility of the bacteria or of functionalized microspheres can be reconstituted in vitro from only five pure proteins. Movement results from the regulated site-directed treadmilling of actin filaments, consistent with observations of actin dynamics in living motile cells and with the biochemical properties of the components of the synthetic motility medium. PMID- 11379634 TI - Secondary hyperparathyroidism: evidence for an association with papillary thyroid cancer. AB - The association between primary hyperparathyroidism and nonmedullary thyroid malignancies is well known. There is also, however, some evidence for an association between secondary hyperparathyroidism (SHPT) and thyroid cancer. We report three patients in whom invasive papillary thyroid carcinoma (PTC) was diagnosed before (one case) or at the time of (two cases) parathyroidectomy for SHPT. Three women (ages 23, 54, and 64 years) presented with bone pain and pruritus typical of SHPT. All three patients had biopsy-proven parathyroid bone disease and elevated parathormone levels (664, 1674, and 2051 pg/mL). All underwent subtotal parathyroidectomy and total thyroidectomy without complications. Pathology revealed diffuse parathyroid hyperplasia with multifocal invasive papillary thyroid carcinoma (two cases) and follicular variant of papillary thyroid carcinoma (one case). Two cases were associated with metastatic disease to local lymph nodes. The patients received adjuvant radioactive 131I, and remained tumor free 24 to 36 months after surgery with complete resolution of SHPT. We conclude: 1) PTC may accompany SHPT, 2) PTCs associated with SHPT may be locally aggressive although usually they are early tumors, 3) surgeons need to have an index of suspicion for thyroid tumor when operating on patients with SHPT, and 4) routine removal of the thymus as part of the operation for SHPT may have a secondary benefit in diagnosing PTC in the occasional patient. PMID- 11379635 TI - Intraductal papillary-mucinous tumor of the pancreas: assessing the grade of malignancy from natural history. AB - Intraductal papillary-mucinous tumor of the pancreas is a spectrum of conditions ranging from benign to malignant, and very few papers have referred to the natural history of this disease. In this communication the indicators of malignancy were examined from a viewpoint of natural history. Follow-up computed tomographies (CTs) more than 6 months after the diagnosis were reviewed in 17 Japanese patients with intraductal papillary-mucinous tumor of the pancreas. They were divided into two groups by the presence or absence of morphological progressive changes by the follow-up CTs, and the clinicopathological features were compared between the two groups to examine possible malignant indicators. The 17 patients consisted of seven patients in the no-change group and ten in the progressive group. The distribution of the patients was not different with regard to age; gender; or presence or absence of pancreatitis, diabetes mellitus, or unique findings of the ampulla of Vater between the two groups. The dilatation of the main pancreatic duct (> or = 3 mm) was more frequent in the progressive group: (eight of ten patients; 80%) than in the no-change group (two of seven patients; 29%) (P = 0.03). Six (86%) of the seven tumors in the no-change group were located in the branch duct, whereas five (50%) of the ten in the progressive group were situated in the main pancreatic duct. Histopathologic diagnoses of the resected specimens of the four in the no-change group examined were intraductal papillary-mucinous adenoma in three and adenoma with moderate dysplasia in one, whereas the diagnoses in the six in the progressive group examined were adenoma in two, adenoma with moderate dysplasia in two, and carcinoma (invasive) in two. The patients with intraductal papillary-mucinous tumor of the pancreas with a dilatation of the main pancreatic duct at the time of diagnosis should be followed up more carefully than those without dilatation. Once progressive morphological changes are detected by the follow-up CTs surgical resection should be considered because of possible malignancy. PMID- 11379636 TI - Abdominal wall penetration by a police "bean bag". AB - Police departments across the country now have at their disposal a "nonlethal" weapon known as a "bean bag" shotgun. The bean bags, which come loaded in standard shotgun shells, are missiles intended to disable dangerous suspects without having to resort to lethal force. They were specifically designed not to create penetrating injuries. Here we present a case of abdominal wall penetration by a police bean bag. To our knowledge this is the first such reported case of this type of injury. PMID- 11379637 TI - Liver infarction after laparoscopic cholecystectomy injury to the right hepatic artery and portal vein. AB - A 66-year-old woman presented with fever and right upper quadrant pain 5 weeks after laparoscopic cholecystectomy. Angiogram revealed occlusion of the right hepatic artery and right portal vein which necessitated a right hepatic lobectomy. To our knowledge this has not been previously reported. The patient recovered uneventfully. PMID- 11379638 TI - Neoadjuvant chemotherapy for Chinese women with locally advanced breast cancer. AB - Locally advanced breast cancer carries a poor prognosis and is still prevalent in developing countries. The current management usually involves administration of neoadjuvant chemotherapy (NCT). From March 1990 through December 1997, 173 Chinese patients with tumor size greater than 4 cm were treated; 38 received NCT and the other 135 postoperative adjuvant chemotherapy. The regimens for NCT were FEC (5-fluorouracil 600 mg/m2, epirubicin 50 mg/m2, and cyclophosphamide 600 mg/m2) for 29 patients and Adriamycin 75 mg/m2 for the rest of the group. Postoperatively the NCT patients received the standard CMF regimen (oral cyclophosphamide 100 mg/m2 for 14 days and intravenous methotrexate 40 mg/m2 and 5-fluorouracil 600 mg/m2 on days one and eight of each cycle). The postoperative adjuvant chemotherapy group received only the CMF regimen. Tumor response after NCT was measured clinically and histologically. The response rate was 75 per cent with 13.2 per cent being complete response. Although there is no difference in response rate the actual reduction in size was greater for patients receiving Adriamycin than FEC (P = 0.001). The only predictive factor of response to NCT was the type of chemotherapy administered. None of the tumor characteristics such as size, nodal status, histological grading, lymphovascular permeation, hormonal receptor status, and c-erb-B2 expression were found to be significant. The overall 5-year probability of survival was 0.44, and there was no difference between groups. The factor important for prognosis was axillary nodal status on histology. The use of NCT did not improve outcome. In summary our results showed that NCT was feasible for Chinese women and good response could be achieved. However, it is the axillary nodal status that determines the final outcome. PMID- 11379639 TI - Are complications of subtotal colectomy with ileorectal anastomosis related to the original disease? AB - The aim of this study was to compare the morbidity of subtotal colectomy with ileorectal anastomosis performed for colonic inertia, Crohn's disease, familial adenomatous polyposis, and colorectal neoplasia. A retrospective review of all patients who underwent elective colectomy with ileorectal anastomosis between June 1988 and November 1996 was performed. The patients were divided into three groups: Group I, colonic inertia; Group II, Crohn's disease; and Group III, Familial Adenomatous Polyposis or other neoplasia. Outcome factors studied included the frequency of bowel movements, the incidence of small bowel obstruction, and the incidence of anastomotic leakage. Other factors assessed included operative time, intraoperative blood loss, length of hospitalization, level of ileorectal anastomosis, time of first bowel movement, and whether the operation was undertaken in one or two stages. Statistical analysis was undertaken by using the chi-square test and the Mann-Whitney U exact test. All 48 patients in Group I were operated on in one stage. In Group II (30 patients) 15 patients were operated on in one stage, eight patients had a preliminary Hartmann's operation and then ileorectal anastomosis, and seven patients underwent subtotal colectomy with both an ileorectal anastomosis and a proximal loop ileostomy. In Group III (22 patients) 20 patients underwent a one-stage operation whereas two patients underwent a subtotal colectomy with ileorectal anastomosis and proximal loop ileostomy. The median ages were 47.0 years in Group I, 43.8 in Group II, and 53.3 in Group III. Small bowel obstruction occurred in five patients (10%) in Group I, four patients (13.3%) in Group II, and four patients (18%) in Group III. The anastomotic leak rate was 4.2% (two patients) in Group I, 1% (three patients) in Group II, and 0% in Group III (P < 0.05). At the follow up interview after surgery, the mean number of bowel movements per day 6 months after surgery was 5.4 in Group I, 7.2 in Group II, and 5.6 in Group III, (P < 0.05, Group II vs Group I or Group III). Operative time in Group III was significantly longer than in the other two groups (P = 0.004). No statistically significant differences were found among the three groups relative to blood loss, hospitalization, or timing of first bowel movement. This study failed to identify any differences in either immediate perioperative outcome or morbidity or intermediate-term function in patients undergoing ileorectal anastomosis regardless of diagnosis. The overall rate of small bowel obstruction was 13 per cent with no significant differences among the three groups. Lastly although the anastomotic leak rate was not significantly higher in patients with Crohn's disease it was higher in the group with ileostomy and ileorectal anastomosis, which highlights a potential advantage of performance of this procedure in two stages in selected patients of this patient population. PMID- 11379640 TI - Closure of the abdominal midline fascia: meta-analysis delineates the optimal technique. AB - The current surgical literature has not clearly demonstrated an optimal technique for abdominal closure. Prospective randomized studies published between 1980 and 1998 were analyzed and the relevant data derived from those studies were pooled for statistical evaluation. The outcome variables of dehiscence, infection, hernia formation, suture sinus formation, and pain were studied and the probability of their occurrence in association with different techniques was calculated. In relation to the outcome features of dehiscence and infection no statistically significant difference was seen when absorbable suture material was compared with nonabsorbable material. In regard to the probability of hernia formation no statistically significant difference was seen when monofilament absorbable material was compared with nonabsorbable material. There was, however, a higher incidence of hernia formation when braided absorbable suture material was used. In addition there was a higher incidence of incision pain and suture sinus formation when nonabsorbable suture material was used. Absorbable monofilament suture material is superior to both absorbable braided and nonabsorbable suture for abdominal fascial closure. A continuous mass (all-layer) closure with absorbable monofilament suture material is the optimal technique for fascial closure after laparotomy. PMID- 11379641 TI - Should trauma surgeons render definitive vascular repair in peripheral vascular injuries? AB - Our hypothesis is that in an established Level I trauma center general trauma surgeons should repair peripheral vascular injuries even in stable patients when there is time for a vascular consult. We reviewed all penetrating peripheral vascular injuries in stable patients operated on by nine experienced general trauma surgeons (1993-1996). Outcome measures were amputation, nerve damage, and vascular complications. There were 43 patients with 44 peripheral vascular injuries identified. Sixty per cent were from stab wounds. There were 27 arterial injuries (carotid four, subclavian one, vertebral two, axillary three, brachial eight, ulnar one, radial two, femoral five, and anterior tibial one). There were three venous injuries (one each subclavian, axillary, and popliteal). There were 14 combined injuries (vertebral two, femoral nine, and popliteal three). There were no mortalities. Morbidity was limited to patients with lower extremity injuries. In the nine patients with combined femoral vessel injury there were three complications (nerve damage, thrombosed arterial repair, and thrombosed venous repair). In the four patients with popliteal venous injuries there were two complications, both venous thrombosis. Our early arterial patency rate was 97.6 per cent. These data support the hypothesis that general surgeons with trauma experience can provide effective treatment of peripheral vascular injuries. The significance of these findings in improving the image of trauma surgery as a career is discussed. PMID- 11379642 TI - Histoplasmosis as an isolated liver lesion: review and surgical therapy. AB - Histoplasmosis is the most common cause of fungal infection in the Ohio River Valley of the United States. Ninety-nine per cent of patients exposed to histoplasmosis develop only subclinical infections. Liver involvement is common in disseminated histoplasmosis, which usually originates in the lungs. There has been only one prior case described in the literature of histoplasmosis presenting as an isolated liver mass. We report a rare case that presented as a solitary right-sided liver lesion invading the diaphragm, with review of the literature for therapy of histoplasmosis of the liver. PMID- 11379643 TI - Visceral artery aneurysms: experience in a tertiary-care center. AB - Visceral artery aneurysms (VAAs) often rupture and cause serious morbidity or death. The purpose of this study was to identify conditions associated with VAA in a series of 30 patients treated at our institution from 1988 through 1998. Demographics, types of aneurysms, associated conditions, diagnoses, treatments, and outcomes were recorded and analyzed. Thirty patients (16 men and 14 women) with VAA were identified. The arteries involved were splenic (eight), renal (ten), hepatic (nine), hypogastric (one), celiac (one), and pancreaticoduodenal (one). Five of eight (63%) splenic artery aneurysms occurred in women; however, gender was not a factor in other aneurysmal groups. Splenic artery aneurysm also was associated with cirrhosis in four of the eight (50%) patients. Five of the nine (56%) hepatic artery aneurysms were associated with cirrhosis; two of these were pseudoaneurysms that occurred after liver transplantation. Five of ten (50%) renal artery aneurysms were associated with juxtarenal abdominal aortic aneurysms. Celiac and pancreaticoduodenal aneurysms were associated with gastrointestinal bleeding. Treatments included surgery (19), embolization (eight), and observation alone (three). These data demonstrate that association with other conditions varies according to subgroups of VAA. Despite advances in diagnosis and therapy the heterogeneity of VAA suggests that management must remain individualized. PMID- 11379644 TI - Digital rectal examination for trauma: does every patient need one? AB - The digital rectal examination is widely accepted as an essential component in the initial assessment of trauma. However, no data have been published that justify its routine use in all seriously injured patients. The objective of this study was to determine what if any impact on subsequent treatment and management decisions the initial digital rectal examination had on injured patients arriving at our emergency department (ED). We conducted a prospective observational study of all injured patients arriving at a Level II trauma center over a period of 6 months. A digital rectal examination was performed on all patients during the secondary survey phase of their initial evaluation shortly after arrival to the ED. The results of the rectal examination were noted for each patient with particular attention placed on the presence or absence of gross blood, Hemoccult result, prostatic examination, rectal vault integrity, and rectal sphincter tone. In addition the patient's hemodynamic parameters while in the ED and the injuries that were sustained were noted, as was their final disposition. Four hundred twenty-three patients were admitted to the ED after sustaining serious injuries. The mean Injury Severity Score was 9.96. The prostatic examination was normal in more than 99 per cent of patients; no high-riding or nonpalpable prostate glands were noted. Twenty-two patients (5.2%) were Hemoccult positive, but in none of these cases did the presence of occult blood in the stool lead to a change in the initial management or diagnostic approach. Three patients (0.7%) with penetrating injuries to the perineal/pelvic area had gross blood on digital rectal examination that prompted operative exploration to rule out a lower gastrointestinal injury. All three had rectal injuries confirmed at surgery. Rectal sphincter tone was normal in 406 (96%) patients, weak in 17 (4%), and absent in none. The only patient in whom the sphincter tone influenced management was an individual complaining of complete paralysis after a blunt mechanism of injury. He had normal rectal sphincter tone and admitted to malingering shortly thereafter. Overall the rectal examination influenced therapeutic decision making in five cases (1.2%). The digital rectal examination is unlikely to affect initial management when applied indiscriminately to all seriously injured patients during the secondary survey. Patients in whom the rectal examination may have a higher probability of influencing management are those with penetrating injuries in proximity to the lower gastrointestinal tract, questionable spinal cord damage, and severe pelvic fractures with potential urethral disruption or open fractures in continuity with the rectal vault. The Hemoccult test does not add useful information and should be discontinued as part of the secondary survey of injured patients. PMID- 11379645 TI - Long-term results of hepaticojejunostomy for hepatolithiasis. AB - The results of a hepaticojejunostomy as a biliary-enteric bypass for benign disease are usually excellent. On the other hand, hepatolithiasis features a high rate of residual and recurrent stones with cholangitis after surgery. This study aims to evaluate the long-term results of a hepaticojejunostomy for hepatolithiasis regarding both the degree of the occurrence of postoperative cholangitis and the outcome. The clinical records of 159 patients with hepatolithiasis who underwent surgical treatment over a 23-year period were also retrospectively reviewed. Ninety-four of 159 patients underwent a hepatecetomy and 65 patients were subjected to liver-preserving surgery by means of intra- and postoperative endoscopic lithotripsy. In addition 72 patients underwent a hepaticojejunostomy. The rate of residual or recurrent stones was 31.4 per cent after complete stone removal. Twenty-two (30.6%) of the 72 patients developed some kind of cholangitis. This rate was significantly higher than that (three of 87 patients) of the non-biliary-enteric anastomosis group regarding the occurrence of biliary complications. We conclude that the use of a hepaticojejunostomy for patients with possible residual stones or intrahepatic bile duct lesions remains controversial. PMID- 11379647 TI - Abdominal compartment syndrome with massive lower-extremity edema caused by colonic obstruction and distention. AB - Abdominal compartment syndrome is a well-described condition in which increased intra-abdominal pressure causes various physiologic derangements with adverse effects on cardiac, pulmonary, and renal function. A patient presented with radiation-induced distal colonic obstruction, abdominal distention, and severe bilateral leg edema. We performed a diverting transverse loop colostomy as treatment for her obstruction. This resulted in massive, spontaneous diuresis with complete resolution of her lower-limb edema. Abdominal compartment syndrome due to colonic obstruction can contribute to the development of lower-extremity edema. Colon decompression with reduction of intra-abdominal pressure can lead to resolution of edema in this situation. PMID- 11379648 TI - Isolated colon cancer metastasis to the spleen. AB - Splenic metastases from colon carcinoma are rare. If present they generally occur in concert with disseminated disease. Six cases have been previously reported. The patient presented here is a 51-year-old black man who presented with an enterocutaneous fistula as a complication from prior pelvic radiation. Workup included an abdominal CT and needle biopsy, which confirmed the splenic metastasis from a sigmoid colon cancer 6 years after the original diagnosis. The patient had an unevenful splenectomy. Although no long-term follow-up data are as yet available splenectomy including removal of hilar lymph nodes appears to be the treatment of choice. A case report and review of the literature are presented. PMID- 11379646 TI - A comparison between topical and infiltrative bupivacaine and intravenous meperidine for postoperative analgesia after inguinal herniorrhaphy. AB - The purpose of the present study is to compare postoperative analgesia offered by the simple instillation of local anesthetic on the surgical wound, its infiltration with the same local anesthetic, and the use of an intravenous opioid. Sixty patients were divided into the three analgesia groups to be studied: instillation of local anesthetic (Group I), injection of local anesthetic (Group II), and intravenous opioid (Group III). The pain was quantified using the visual analogue scale. It was observed that there was better analgesia in Groups I and II during the first 6 hours postoperatively as compared with Group III (P < 0.0001). At the end of the 12 hours the three modes of analgesia proved comparable. However, after 24 hours there was better analgesic development in Group I, whereas Group II had greater postoperative morbidity. We conclude that the instillation of local anesthesia provides analgesia during the immediate postoperative period comparable to local infiltration using the same anesthetic. Both regional analgesia methods are more effective analgesics during the first 6 hours than are intravenous opioids. Furthermore the simple instillation of local anesthetic allows better analgesic evolution of the surgical wound after the first 24 hours considering the lower rate of resulting complications. PMID- 11379649 TI - Diverticulitis in the young patient. AB - Although predominantly a disease in older adults diverticulitis does affect younger patients. The disease has been described as not only rare but virulent by some authors, and a young patient age is considered to be a relative indication for early sugery. The goal of this study was to evaluate the experience of the Louisiana State University Health Sciences Center-Shreveport and affiliated hospitals with diverticulitis in young patients. This study was a retrospective chart review of 22 patients with diverticulitis age 40 years and younger over the past 20 years. Inclusion criteria were either a diagnosis of diverticulitis confirmed at surgery or positive CT findings and/or a positive contrast enema. The mean age in this study was 32.1 years (range 16-40). All 22 patients presented with abdominal pain. The next most common symptom was nausea and/or vomiting in 45 per cent followed by fever and chills in 36 per cent. Twelve patients had abdominal CTs on admission, and 87 per cent had positive findings. Eighteen patients underwent an operation. Four patients were treated nonoperatively. Nineteen patients had diverticulitis of the sigmoid colon. The remaining three had right-sided diverticulitis. Two patients underwent right hemicolectomy, and one underwent cecectomy. Of the 15 patients with sigmoid diverticulitis 12 (80%) underwent a two-stage procedure of sigmoid colectomy, end colostomy, and Hartmann's pouch. Three patients (20%) underwent a one-stage procedure of sigmoid colectomy and primary anastomosis. Two of three patients undergoing a one-stage procedure required reoperation. Postoperative complications occurred in 10 of 18 patients for an overall incidence of 56 per cent. Two of these patients had septic complications. Both of these patients had a delay in time from admission until operation: one for 7 days and the other for 10 days. There was one death in the series. Colostomy closure was performed successfully in nine of 12 (75%) patients. The mean time interval before closure was 7.7 months, (range 3-14). Patients with two-stage procedures on initial admission fared better than those with one-stage procedures. The overall mortality was 4.5 per cent. There was a high overall complication rate of 56 per cent in patients undergoing an operation. Two patients who had a delay in time from admission to operation had septic complications. Early surgical intervention should be considered in this clinical setting. In summary, although rare, diverticulitis in the young patient is often a fulminant illness requiring operation early in the disease process. PMID- 11379650 TI - Outcome of surgical treatment of chronic pancreatitis associated with sphincter of Oddi dysfunction. AB - The clinical management of patients with chronic pancreatitis (CP) associated with sphincter of Oddi dysfunction (SOD) presents many challenges. The aim of this study was to evaluate patient outcome after surgical management of CP associated with SOD intractable to medical management. The records of patients with CP and SOD who underwent surgical treatment between 1994 and 1998 were retrospectively reviewed and analyzed. Manometry of biliary and pancreatic ducts was performed. Basal pressures were considered abnormal if > or = 40 mm Hg for at least 30 seconds. Endoscopic retrograde cholangiopancreatography, endoscopic ultrasound, and clinical criteria were utilized in the diagnosis of CP. Quality of-life issues were assessed. Twenty-nine patients were identified (21 women and eight men) with a mean age of 43.3 years (range 24-54). Mean basal biliary and pancreatic sphincter pressures were 155.1 and 90.4 mm Hg respectively. Chronic pancreatitis was graded as mild in nine patients, moderate in six, severe in two, and normal or equivocal in 12 patients according to the Cambridge classification. A Whipple procedure was performed in 17 (59%) patients, lateral pancreaticojejunostomy in nine (31%), and distal resections or a combination of procedures in three (10%). The morbidity and mortality rates were 21 and 0 per cent respectively. Mean follow-up was 30 months (range 3-48). Pain relief ranging from fair to excellent was seen in 83 per cent of patients with pain scores decreasing from an average of 9 (scale 1-10) before surgery to 3 postoperatively. Seventy per cent maintained their weight, 45 per cent continued to require pancreatic enzyme supplementation, and there were no changes in the status of diabetes. Rehospitalizations for recurrent pancreatitis or persistent pain were necessary in 24 per cent of patients. Surgical management of patients with CP and SOD who fail medical management is safe and effective in most patients. Operative morbidity and mortality are low, and the majority of patients have improvement in pain, although some require rehospitalization for recurrent pancreatitis and chronic pain. PMID- 11379651 TI - Talc pleurodesis: a new technique. AB - Pleurodesis is a commonly used option in the management of malignant pleural effusion. Although several chemical agents are used talc has only recently been advocated as the sclerosing agent of choice. Talc can be administered via open thoracotomy or tube thoracostomy, but thoracoscopy offers an excellent alternative to these approaches. We describe a new technique of thoracoscopically applied talc pleurodesis that is easy, inexpensive, and reliable. PMID- 11379652 TI - Anal carcinoma arising from condyloma acuminata. AB - Condyloma acuminata is a common anorectal condition that frequently requires surgical evaluation and treatment. We have noted an increased incidence of anal carcinoma in patients with condyloma acuminata. The purpose of this study is to review the incidence of malignant transformation of condyloma in our recent experience. We conducted a 5-year retrospective review of patients with condyloma acuminata treated at a university medical center that serves as a major referral center for the state. From May 1994 through May 1999 257 patients were treated for anal condyloma. During the same time period 74 patients were diagnosed with squamous cell carcinoma of the anus; nine of these patients also had condyloma acuminata (12.2% of patients with anal carcinoma). All nine were immunosuppressed by illness and/or medication. The extent of carcinoma at diagnosis ranged from stage 0 (carcinoma in situ) to stage IVb. Overall 3.5 per cent of patients with condyloma acuminata also had squamous cell carcinoma of the anus. One patient with stage IVb disease died shortly after initial evaluation. Two patients with advanced disease required extensive surgical intervention and had complex postoperative courses. Malignant transformation of condyloma acuminata may be increasing in incidence. This disease progression can be insidious and may be fatal. Screening of high-risk patients might be of value, and more aggressive early management of condyloma may prevent the development of malignancy. PMID- 11379653 TI - Iatrogenic and noniatrogenic extrahepatic biliary tract injuries: a multi institutional review. AB - Traumatic and iatrogenic extrahepatic biliary tract injuries are rare but may lead to exceedingly morbid complications. Traumatic extrahepatic biliary tract injuries represent less than 1 per cent of all traumatic injuries. Iatrogenic injuries result in 0.2 to 1 per cent of laparoscopic or open cholecystectomies. The objective of this study was to review the incidence of biliary tract injuries -iatrogenic as well as traumatic--and their subsequent management. A multi institutional chart review was done including Louisiana State University Health Sciences Center (LSUHSC)-Shreveport, LSUHSC-Monroe, and Richland Parish medical centers. Charts were reviewed for patients with iatrogenic biliary tract injuries and those with biliary tract injuries related to noniatrogenic trauma. The etiology of the biliary tract injury, symptoms of injury, pertinent laboratory and radiologic studies, injury-to-diagnosis time, type of biliary tract injury, injury management, days hospitalized, intensive care unit stay, and complications were reviewed. There are 1500 trauma patients admitted to LSUMC-Shreveport each year. The incidence of biliary tract injury in trauma patients admitted to LSUMC is 0.1 per cent. Traumatic injuries were classified according to the injury scale by Mattox et al. (Trauma 1996; Vol 515). There were five Type II, four Type IV, and two Type V injuries. Five patients underwent cholecystectomy, three had endoscopic retrograde cholangiopancreatography with stent placement, and two had choledochojejunostomy; one patient died from associated injuries. There were no complications of repair. Approximately 220 cholecystectomies are done at LSUMC Shreveport each year. Eighty-eight per cent are laparoscopic, and 12 per cent are open. The incidence of iatrogenic biliary tract injuries at LSUMC-Shreveport during the past 8 years was 0.2 per cent. Immediate diagnosis of iatrogenic injuries was made in five of 17 cases and eight of 11 trauma cases. Laparoscopic injuries were classified by the Way injury classification (Stewart L, Way LW. Arch Surg 1995;130:1123). There were one Type I, one Type II, and nine Type III injuries. Treatment included suturing of the laceration (n = 1), hepaticojejunostomy (n = 8), and primary repair (n = 2). Open injuries were classified using the Bismuth classification. There were one Type I and three Type III injuries. All were treated with hepaticojejunostomy. There were two iatrogenic injuries unrelated to cholecystectomy. One patient suffered a perforation of the gallbladder during laparoscopic nephrectomy. This patient subsequently underwent cholecystectomy and has done well. The second patient suffered ligation of the intraduodenal portion of the common bile duct during hemigastrectomy and oversewing of a duodenal ulcer. This patient underwent hepaticojejunostomy and has done well. Complications of iatrogenic injury repair included leaking of a repaired laceration (n = 1), failed hepaticojejunostomy (n = 1), and an anastomotic stricture after hepaticojejunostomy (n = 1). Laparoscopic injuries by LSUMC hospitals is 0.2 per cent. Extrahepatic biliary tract injuries resulting from open cholecystectomy were diagnosed later than those occurring during laparoscopic cholecystectomy and were most likely to result in stricture formation. Repair of Way Type II and III injuries is associated with a higher complication rate. Hepaticojejunostomy has a complication rate of 15 per cent. Minor common duct lacerations are amenable to conservative therapy with oversewing and/or endoscopic retrograde cholangiopancreatography with stent placement. Repair of extrahepatic biliary tract injuries with hepaticojejunostomy at a level of good blood supply remains our gold standard for treatment of more severe injuries and strictures. PMID- 11379654 TI - Immediate and long-term outcomes after lateral pancreaticojejunostomy for chronic pancreatitis. AB - Lateral pancreaticojejunostomy (LPJ) is the recommended surgical treatment of intractable pain from chronic pancreatitis (CP) with obstruction and ductal dilatation. This study evaluated the etiology, morbidity, mortality, hospital costs, and quality of life (QL) for patients with LPJ for CP. Medical records of 60 patients undergoing LPJ for CP between 1988 and 1996 were reviewed. Long-term QL was assessed by the Short Form 36 Health Survey and analyzed against control populations of patients who underwent pancreatic debridement for necrosis and patients with laparoscopic cholecystectomy for cholelithiasis. CP etiologies included 52 per cent alcoholic, 28 per cent idiopathic, 13 per cent pancreatic divisum, and 7 per cent familial pancreatitis. Peri- and postoperative morbidity and mortality were 25 and 0 per cent respectively. Average hospital cost was $13,530 with mean postoperative hospital stay of 12.1 days. Overall physical and mental QL were diminished compared with both the debridement group and cholecystectomy group with particular detriments in areas of physical role (P < 0.05), bodily pain (P < 0.001), social function (P < 0.001), and mental health (P < 0.001). We conclude that LPJ for CP is a relatively safe procedure with low morbidity and mortality but results in a significantly diminished long-term QL relative to other surgical patients with pancreatic or biliary disease. This difference prevails in both physical and mental aspects of health. PMID- 11379655 TI - Management of perianal sepsis in immunosuppressed patients. AB - Despite improvements in the supportive care of immunosuppressed patients controversy still surrounds the surgical management and outcome of anorectal sepsis in these patients. We reviewed 83 immunocompromised patients with diagnosis of perianal sepsis from 1995 to 1997. Sixty-six patients (80%) were followed for a mean of 15 months. Mean age was 44 years and 76 per cent were males. Twenty-eight per cent were HIV+, 34 per cent had inflammatory bowel disease on steroids, 20 per cent had malignancies, and 18 per cent had diabetes. Twenty-eight per cent had anal fistula, 2 per cent had perianal abscess, and 40 per cent had both. Primary sites of fistula were: transsphincteric (38%), intersphincteric (33%), superficial (20%), and suprasphincteric (3%), and multiple tracks (6%). Horseshoeing was present in 14 per cent of cases. The most commonly practiced surgical procedures were primary fistulotomy (n = 23) and fistulotomy plus drainage (n = 28). Seven patients underwent fistulotomy and ostomy and eight patients were treated with fistulectomy plus drainage. Most wounds (91%) healed within 8 weeks. Incontinence (6%) and recurrence (7%) were the most commonly observed complications. These results are similar to those seen in the general population. Perianal sepsis can be safely managed in immunocompromised patients, with high rates of healing and low complication rates. An aggressive sphincter-preserving approach in the management of these patients may be undertaken. PMID- 11379656 TI - Surgical management for chronic pancreatitis. AB - This study reviewed the results of surgery for chronic pancreatitis. We also attempted to identify any factors that may influence outcome. A 10-year retrospective chart review was carried out on all patients undergoing surgery for the diagnosis of chronic pancreatitis. Twenty-three patients were identified. Alcohol was the most common etiology, but other causes were identified. All but two patients had abnormal ductal anatomy on endoscopic retrograde cholangiopancreatography. A total of 23 patients underwent six different operations. There were five complications and no perioperative deaths. Only one patient had to be readmitted for pancreatitis during follow-up. The majority of patients reported some improvement with their pain. Patients who continued to use alcohol had the worst results in regard to weight gain and pain control. The results of our study are consistent with the current literature in regard to morbidity and mortality. Surgical treatment had minimal effect on endocrine and exocrine function. Weight loss was avoided in the majority of patients. Addition of biliary bypass to the Puestow procedure did not increase morbidity. Poorest results were obtained in patients who continued to use alcohol. A basic algorithm for management of this disease process is given. PMID- 11379658 TI - Carcinoma arising in an amputation stump. AB - We describe a 62-year-old diabetic man who presented with squamous cell carcinoma on his right thigh amputation stump 52 years after the procedure. The potential relationship of the chronic irritation by the artificial leg and the malignant transformation of the healing scar as well as optimal treatment options of scar tissue carcinomas are reviewed. PMID- 11379659 TI - Extraocular phototransduction and circadian timing systems in vertebrates. AB - It is widely accepted that, for organisms with eyes, the daily regulation of circadian rhythms is made possible by light transduction through those organs. Yet, it has been demonstrated repeatedly in recent years that ocular light receptors that mediate vision, at least in mammals, are not the same photoreceptors involved in circadian regulation. Moreover, it has been recognized for many years that circadian regulation can occur in organisms without eyes. In fact, extraocular circadian phototransduction (EOCP) appears to be a phylogenetic rule for the vast majority of species. EOCP has been reported in every nonmammalian species studied to date. In mammals, however, the story is very different. This paper presents findings from studies that have examined specifically the capacity for EOCP in vertebrate species. In addition, the literature addressing noncircadian aspects of extraocular phototransduction is briefly discussed. Finally, possible mechanisms underlying EOCP are discussed, as are some of the implications of the presence, or absence, of EOCP across phylogeny. PMID- 11379657 TI - The dramatic presentation of colonic lipomata: report of two cases and review of the literature. AB - Colonic lipoma with a dramatic presentation requiring urgent operation is a rare occurrence. We report two such cases in conjunction with a review of the literature on colonic lipomata. Clinicopathologic features of two patients who required urgent resection were studied. The preoperative diagnosis of colonic lipoma was suggested on imaging study in one case. A MEDLINE search was conducted with a special goal of revealing cases with a dramatic presentation. One patient presented with rectal bleeding and intussusception related to a partially infarcted 4.5-cm submucosal lipoma of the lower descending colon. The second patient presented with intestinal obstruction related to a near-totally infarcted 6-cm submucosal lipoma at the splenic flexure. In both cases a florid reactive vascular and fibro-/myofibroblastic proliferation and associated hyperplastic mucosal pattern were present at the base and edge of the lipoma. Among 275 previously reported cases of colonic lipoma 28 patients had a dramatic presentation with pain and/or rectal bleeding being the most significant prodromal symptom. In this subset the lipomas tended to be larger, frequently had associated marked necrosis/ulceration, and were less likely to be located in the ascending colon/cecum. Whereas colonic lipomas are relatively common occasional cases present dramatically with massive bleeding, intussusception, or even perforation for which emergency operation is required. Such lipomas usually reveal marked ischemic changes. PMID- 11379660 TI - The birth of chronobiology: Julien Joseph Virey 1814. AB - Julien-Joseph Virey (1775-1846) held the position of pharmacist-in-chief at the Val-de-Grace, a military hospital. He was an innovative pharmacist, naturalist, anthropologist, and philosopher and a prolific author. His writings encompassed a wide range of topics, although many of his ideas were sometimes harshly questioned. Interest in Virey's work today stems from renewed appreciation of his doctoral thesis in medicine, which was completed in 1814 in Paris and was the first devoted to biological rhythms. Virey envisioned biological rhythms to be innate in origin and controlled by living clocks entrained by periodic environmental changes, such as the day-night alternation in light and darkness. He also reported that the effects of drugs vary according to their administration time. But, above all, he collected and published quantified time series that demonstrated human circadian and annual mortality rhythms. Statistical analysis of Virey's data using modern time series methods confirms his deduction that human mortality exhibits rhythmicity. Comparison of his findings with those derived from analyses of more recent human mortality time series shows the characteristics of these rhythms have changed little since 1807 despite differences in environmental conditions. Virey deserves credit for establishing the field of chronobiology based on his insights and writings. PMID- 11379661 TI - The effect of repeated pulses of light at the same time on period responses of the rat circadian pacemaker. AB - A consequence of simple velocity-based models is that, in response to light pulses, the circadian period should adjust inversely to phase. In addition, because of the interaction of circadian period and phase response, earlier circadian period changes should modify later circadian period changes. The literature contains few mentions of response curves of circadian period responses following light pulses. Rats were exposed to four pulses of light (60 minutes, 1000 lux) at the same circadian time, a minimum of 26 days apart; we assessed period responses and possible bias in the period-response curve. Modulation of circadian period following light-induced phase responses was examined by assessing the period of running wheel activity onset. Phase and circadian period were not consistently found to share an inverse relationship. Moreover, biases in initial period tended to be increased by the experimental protocol regardless of circadian time of pulse. Rats with a short initial (high-velocity) period had a lengthened period, while rats with a long initial period (low velocity) tended to have a reduce period. However, rats with a long initial period were phase delay biased, not phase advance biased. These results do not support a simple velocity model of the pacemaker. PMID- 11379662 TI - Exogenous corticosteroid and shifts of circadian rhythms in hamsters. AB - We tested the hypothesis that glucocorticoid stimulation mediates the effect of exercise on circadian clock resetting in hamsters. We injected animals with 1 and 5 mg dexamethasone--a potent glucocorticoid agonist--at zeitgeber time (ZT) 4 and ZT6, circadian phases at which vigorous exercise induces maximal phase advances of about 3 h. Neither dose of dexamethasone induced phase shifts that were significantly larger than those induced by injections of saline vehicle at either of the phases tested. Some animals, however, showed quite large and consistent phase shifts to repeated injections whether with saline or dexamethasone, such that there was a statistically significant correlation between individuals' responses to the two treatments. The data indicate no role for increased glucocorticoid activity in mediating the effects of exercise on circadian phase shifting, but suggest a modest role for nonspecific stimulation, independent of exercise, in inducing phase shifts at ZT4-ZT6. PMID- 11379663 TI - Reduced baroreflex sensitivity and blunted endogenous nitric oxide synthesis precede the development of hypertension in TGR(mREN2)27 rats. AB - Transgenic TGR(mREN2)27 (TGR) rats are an animal model of fulminant hypertension characterized by an inverse circadian blood pressure profile. The present study addressed the contribution of nitric oxide (NO) synthesis and baroreflex function to hypertension and the inverse blood pressure pattern. NO synthesis was measured at four different times of day indirectly by excretion of NO metabolites (NOx: NO2- and NO3-) in the urine of 5- and 11-week-old TGR and Sprague-Dawley (SPRD) controls. Blood pressure, heart rate, and motor activity were recorded in age matched rats of both strains using an implantable telemetry system. Beat-to-beat recording of blood pressure and pulse interval was performed hourly in 6-week-old animals over 24 h. From these data, baroreflex sensitivity (BRS) was calculated by linear regression of spontaneous fluctuations of blood pressure and corresponding changes of pulse interval. Baroreflex sensitivity was lower in pre hypertensive TGR rats than in SPRD rats, and the reduction was restricted to the daily resting period. In both strains, NOx excretion showed circadian rhythmicity, with peak values during the activity period at night. Interestingly, excretion of NOx was reduced during the resting period in 5-week-old TGR rats prior to the development of hypertension. Impairment of NO synthesis and baroreflex function precede the development of hypertension in TGR rats. The reduction of both parameters was restricted to the resting period and, therefore, could be involved in the development of the inverse circadian blood pressure profile of TGR rats. PMID- 11379664 TI - Temperature profiles, and the effect of sleep on them, in relation to morningness eveningness in healthy female subjects. AB - There were 15 healthy female subjects, differing in their position on the "morningness-eveningness" scale, studied for 7 consecutive days, first while living a sedentary lifestyle and sleeping between midnight and 08:00 and then while undergoing a "constant routine." Rectal temperature was measured at regular intervals throughout this time, and the results were subjected to cosinor analysis both before and after "purification" for the effects of physical activity. Results showed that there was a phase difference in the circadian rhythm of core temperature that was associated with the morningness score, with calculations that "morning types" would be phased earlier than "evening types" by up to about 3 h. This difference in phase (which was also statistically significant when the group was divided by a median split into a "morning group" and an "evening group") could not be attributed to effects of waking activity and existed in spite of the subjects keeping the same sleep-wake schedule. Moreover, it persisted when the subjects' data had been purified and when the data were obtained from the constant routine. That is, there was an endogenous component to this difference in phase of the core temperature. The morning group also showed a greater fall of core temperature during sleep; this was assessed in two ways, the main one being a comparison of constant routine and nychthemeral data sets after correction for any effects of activity. Even though the morning group was sleeping at a later phase of their circadian temperature rhythm than was the evening group, neither group showed a fall of temperature due to sleep that varied with time elapsed since the temperature acrophase. It is concluded that another factor that differs between morning and evening types is responsible for this difference. PMID- 11379665 TI - Circadian rhythmicity of cortisol and body temperature: morningness-eveningness effects. AB - The purpose of this study was to describe and compare the circadian rhythm of body temperature and cortisol, as well as self-reported clock times of sleep onset and offset on weekdays and weekends in 19 healthy adult "larks" (morning chronotypes) and "owls" (evening chronotypes), defined by the Home and Ostberg questionnaire. Day-active subjects entered the General Clinical Research Center, where blood was sampled every 2 h over 38 h for later analysis for cortisol concentration by enzyme immunoassay. Rectal body temperature was measured continuously. Lights were turned off at 22:30 for sleep and turned on at 06:00, when subjects were awakened. The acrophases (peak times) of the cortisol and temperature rhythms occurred 55 minutes (P < or = .05) and 68 minutes (P < .01), respectively, earlier in the morningness group. The amplitude of the cortisol rhythm was lower in the eveningness than in the morningness group (P = n.s.). Subject groups differed on all indices of habitual and preferred timing of sleep and work weekdays and weekends (P = .05-.001). PMID- 11379666 TI - Hypersensitivity of melatonin suppression in response to light in patients with delayed sleep phase syndrome. AB - Patients with delayed sleep phase syndrome (DSPS) experience a chronic mismatch between the usual daily schedule required by the individual's environment and their circadian sleep-wake pattern, resulting in major academic, work, and social problems. Although functional abnormalities of the circadian pacemaker system have been reported in patients with DSPS, the etiology of DSPS has not been fully elucidated. One hypothesis proposed to explain why patients with DSPS fail to synchronize their 24 h sleep-wake cycle to their environment is that they might have reduced sensitivity to environmental time cues, most notably light-dark cycles. Therefore, we compared the sensitivity of melatonin suppression in response to light in patients with DSPS and normal control subjects. Fifteen patients with DSPS and age- and sex-matched healthy controls were studied. As the melatonin secretion rhythm in patients with DSPS was expected to be delayed compared to the controls, the time of peak melatonin secretion was determined in each subject in the first session. In the second session, each subject was exposed to light with an intensity of 1000 lux for 2 h beginning 2 h prior to his or her peak melatonin secretion. Melatonin was measured by radioimmunoassay in saliva sampled every 30 minutes during the period of light exposure. Suppression of the melatonin concentration in saliva was dependent on duration of light exposure. In addition, the suppressive effect of light on the melatonin concentration was significantly greater in patients with DSPS than in control subjects. The results suggest hypersensitivity to nighttime light exposure in patients with this syndrome. Our findings therefore suggest that evening light restriction is important for preventing patients with DSPS from developing a sleep phase delay. PMID- 11379667 TI - Circadian variation of serum leptin in healthy and diabetic men. AB - Leptin, from the Greek leptos, meaning thin (in reference to its ability to reduce body fat stores), is a hormone secreted primarily by adipocytes. At one time, leptin was portrayed as a potential means of combating obesity. Recently, leptin has been identified as a potent inhibitor of bone formation, acting through the central nervous system. Since numerous studies clearly show that bone remodeling is circadian rhythmic with peak activity during sleep, it is of interest to explore circadian variability in serum leptin. Accordingly, circadian characteristics of serum leptin were examined in 7 clinically healthy men and 4 obese men with type II diabetes. Blood samples were collected for 24 h at 3 h intervals beginning at 19:00. The dark (sleep) phase of the light-dark cycle extended from 22:30 to 06:30, with brief awakening for sampling at 01:00 and 04:00. Subjects consumed general hospital meals (2400 calories) at 16:30, 07:30, and 13:30. Serum leptin levels were determined by a R&D Systems enzyme immunoassay technique. Data were analyzed by linear least-squares estimation using the population multiple components method. A statistically significant (P < .018) circadian rhythm modeled by a single 24 h cosine curve characterized the data of each group. The 24 h mean leptin level was statistically greater (P < .001) in the obese diabetic men than in the healthy men (9.47 +/- 0.66 ng/mL vs. 24.07 +/- 1.71 ng/mL, respectively). Higher leptin levels occurred between midnight and roughly 02:30, and lowest leptin levels occurred between noon and the early afternoon. The phasing of this rhythm is similar to the circadian rhythm in bone remodeling previously described. Our results suggest the findings from a single morning blood sampling for leptin may be misleading since it may underestimate the mean 24 h and peak concentrations of the hormone. PMID- 11379668 TI - Nonlinear estimation and statistical testing of periods in nonsinusoidal longitudinal time series with unequidistant observations. AB - The analysis of multiple components is often used to model biological variables that show nonsinusoidal predictable changes of known periods. In general, to anticipate the periods is not easy, and even in cases when we have some a priori information, it is advisable to have a statistical tool to test the chosen periods. In this work, we introduce a statistical procedure to estimate periods of longitudinal series by applying nonlinear regression techniques to the multiple sinusoidal model, as well as to the general linear model. Approximate inferences about the parameters of the model are carried out under the usual hypothesis of normality, independence, and constant variance of the errors. Confidence intervals (CIs) for each individual parameter, as well as for the amplitude-acrophase pair or for any other subgroup of parameters of interest, can be computed. As in the linear analysis of multiple components, it is possible to check the existence of rhythm by means of a zero-amplitude test. The method also allows statistical testing of several hypotheses related to the periods. For example, it is possible to test if the periods are equal to certain values of chronobiologic interest and to check if some components included in the model are harmonically related. On the other hand, when the fitted components have proximal periods, the method allows one to verify if they are modeling the same or different spectral peaks. The method, which was validated by a simulation study for a model of two components and is illustrated by an example of modeling the diastolic blood pressure of two subjects, represents a new step in the development of statistical procedures in chronobiology. PMID- 11379669 TI - An underlying circannual rhythm in seasonal affective disorder? AB - A patient diagnosed with seasonal affective disorder (SAD) carried out prospective ratings of depression weekly for nearly a decade. A winter peak of depression and benzodiazepine intake was documented. However, over the years, the depressive episodes shifted toward spring in an apparent free-running circannual rhythm (periodogram peaks at 53 and 55 weeks). This patient may have an underlying seasonal propensity to depression no longer precisely entrained to environmental cues. PMID- 11379670 TI - A community-based study of seasonal variation in the onset of chronic fatigue syndrome and idiopathic chronic fatigue. AB - One proposed hypothesis regarding the etiology of chronic fatigue syndrome (CFS) is that there is a subgroup of patients in which symptom onset is precipitated by a viral infection. If this is indeed true, then one would anticipate a greater incidence of the emergence of CFS symptoms during months when viral infections occur with the greatest frequency. The current community-based epidemiology study examined the month of symptom onset for 31 patients with CFS and 44 others with idiopathic chronic fatigue (ICF). It was determined that the distribution of the month of illness onset for the CFS and ICF groups was nonrandom, with greater numbers of participants than expected reporting an onset of CFS and ICF during January. PMID- 11379671 TI - What's new about ventilator-associated pneumonia. PMID- 11379672 TI - Is ventilator-associated pneumonia an independent risk factor for death? AB - BACKGROUND: Ventilator-associated pneumonia (VAP) has been implicitly accused of increasing mortality. However, it is not certain that pneumonia is responsible for death or whether fatal outcome is caused by other risk factors for death that exist before the onset of pneumonia. The aim of this study was to evaluate the attributable mortality caused by VAP by performing a matched-paired, case-control study between patients who died and patients who were discharged from the intensive care unit after more than 48 h of mechanical ventilation. METHODS: During the study period, 135 consecutive deaths were included in the case group. Case-control matching criteria were as follows: (1) diagnosis on admission that corresponded to 1 of 11 predefined diagnostic groups; (2) age difference within 10 yr; (3) sex; (4) admission within 1 yr; (5) APACHE II score within 7 points; (6) ventilation of control patients for at least as long as the cases. Precise clinical, radiologic, and microbiologic definitions were used to identify VAP. RESULTS: Analysis was performed on 108 pairs that were matched with 91% of success. There were 39 patients (36.1%) who developed VAP in each group. Multivariate analysis showed that renal failure, bone marrow failure, and treatment with corticosteroids but not VAP were independent risk factors for death. There was no difference observed between cases and controls concerning the clinical and microbiologic diagnostic criteria for pneumonia. CONCLUSION: Ventilator-associated pneumonia does not appear to be an independent risk factor for death. PMID- 11379673 TI - Celiac plexus block: injectate spread and pain relief in patients with regional anatomic distortions. AB - BACKGROUND: The success of the neurolytic celiac plexus block, despite different approaches and methods used, depends on adequate spread of the injectate in the celiac area. This retrospective study was conducted to evaluate the patterns of alcohol spread and pain relief in patients with cancer or therapy-related anatomic distortion of the celiac area. METHODS: From 177 cancer patients who underwent computed tomography (CT)-guided single-needle neurolytic celiac plexus block via an anterior approach, a radiologist, blind to the aim of the study, retrospectively selected 105 patients with abnormal anatomy of the celiac area as judged by CT images obtained before the block. To evaluate CT patterns of neurolytic (mixed with contrast) spread, the celiac area was divided on the frontal plane into four quadrants: upper right and left and lower right and left, as related to the celiac artery. Results were expressed as the number of quadrants into which contrast spread, ie., four, three, two, or one quadrants with contrast. The patterns of contrast spread according to the number of quadrants with anatomic distortion were analyzed. Patient assessment by visual analog scale was reviewed to evaluate the degree of pain relief. Pain relief 30 days after block was considered long-lasting. Pain relief at 30 days after block was analyzed according to the number of quadrants with contrast. RESULTS: Overall, four, three, two, and one quadrants with contrast were observed in 9 (8%), 21 (20%), 49 (47%), and 26 (25%) patients, respectively. An inverse correlation was observed between the number of quadrants with anatomic distortion and the number of quadrants with contrast (P < 0.001). Long-lasting pain relief was noticed in nine of nine patients (100%; 95% confidence interval, 66-100) with contrast in four-quadrants, and in 10 of 21 patients (48%; 95% confidence interval, 26-70) with contrast in 3 quadrants (P < 0.01). None of the 75 patients with contrast in two quadrants or one quadrant experienced long-lasting pain relief. CONCLUSIONS: These findings suggest that, using the single-needle anterior approach, the neurolytic spread in the celiac area is highly hampered by the regional anatomic alterations. It also appears that only a complete (four quadrants) neurolytic spread in the celiac area can guarantee long-lasting analgesia, and that this picture may be obtained in a very limited fraction of patients with regional anatomic alterations. PMID- 11379674 TI - Modeling population pharmacokinetics of lidocaine: should cardiac output be included as a patient factor? AB - BACKGROUND: Inclusion of cardiac output and other physiologic parameters, in addition to or instead of, demographic variables might improve the population pharmacokinetic modeling of lidocaine. METHODS: Thirty-one patients were included in a population pharmacokinetic study of lidocaine. After bolus injection of lidocaine (1 mg/kg), 22 or 10 blood samples per patient were taken from a radial artery. During the experiment, cardiac output was measured using a thoracic electrical bioimpedance method. The following four population pharmacokinetic models were constructed and their performances investigated: a model with no covariates, a model with cardiac output as covariate, a model with demographic covariates, and a model with both cardiac output and demographic characteristics as covariates. Model discrimination was performed with the likelihood ratio test. RESULTS: Inclusion of cardiac output resulted in a significant improvement of the pharmacokinetic model, but inclusion of demographic covariates was even better. However, the best model was obtained by inclusion of both demographic covariates and cardiac output in the pharmacokinetic model. CONCLUSIONS: When population pharmacokinetic models are used for individualization of dosing schedules, physiologic covariates, e.g., cardiac output, can improve their ability to predict the individual kinetics. PMID- 11379675 TI - Intrathecal ropivacaine and clonidine for ambulatory knee arthroscopy: a dose response study. AB - BACKGROUND: The aim of this study was to evaluate the association of a small dose of intrathecal ropivacaine with small doses of intrathecal clonidine for ambulatory surgery. METHODS: One hundred twenty patients, classified as American Society of Anesthesiologists physical status I and scheduled for knee arthroscopy, were studied. Patients were randomly assigned to receive 4 ml of one of the following double-blinded isobaric intrathecal solutions: 8 mg of ropivacaine (group 1; n =30); 8 mg ropivacaine plus 15 microg clonidine (group 2; n =30); 8 mg ropivacaine plus 45 microg clonidine (group 3; n =30); and 8 mg ropivacaine plus 75 microg clonidine (group 4; n =30). The level and duration of sensory anesthesia were recorded, along with the intensity and duration of motor block. Patient and surgeon were interviewed to evaluate the quality of anesthesia. RESULTS: Intrathecal ropivacaine (8 mg alone) produced short sensory anesthesia and motor blockade (132 +/- 38 min and 110 +/- 35 min; mean +/- SD). However, the quality of anesthesia was significantly lower than in any other group (P < 0.05). Ropivacaine (8 mg) plus 75 microg clonidine produced significantly longer sensory and motor anesthesia (195 +/- 40 min and 164 +/- 38 min; P < 0.05). However, this was associated with systemic effects, such as sedation and reduction of arterial blood pressure. Ropivacaine (8 mg) plus 15 microg clonidine did not prolong sensory or motor blockade, afforded high quality anesthesia, and was not associated with detectable systemic effects. CONCLUSION: Small-dose intrathecal clonidine (15 microg) plus 8 mg intrathecal ropivacaine produces adequate and short-lasting anesthesia for knee arthroscopy. PMID- 11379676 TI - Caudal ropivacaine in infants: population pharmacokinetics and plasma concentrations. AB - BACKGROUND: Ropivacaine is a new long-acting amino-amide local anesthetic. However, there are no data on its use in infants. In the current study, the authors investigated the pharmacokinetics of caudal ropivacaine in 30 infants younger than 12 months. METHODS: Two groups of infants (group 1 [n = 15], aged 0 3 months; group 2 [n = 15], aged 3-12 months) were given a caudal bolus dose of 0.2% ropivacaine (2 mg/kg) and a standardized general anesthetic technique. Serial blood samples taken for up to 12 h were analyzed for total and free ropivacaine using high-performance liquid chromatography. Population pharmacokinetic modeling was performed to yield estimates of clearance, volume of distribution, and absorption rate constant. An analysis of covariates on the kinetic parameters also was made. RESULTS: Median maximum free ropivacaine concentration was significantly higher in group 1 (99 micog/l) than in group 2 (38 microg/l) (P = 0.0002), as was the median free fraction of ropivacaine (10% vs. 5%; P = 0.01). Pharmacokinetic variables of the total population were best described by a one-compartment model with first-order absorption. Mean clearance was 0.31 l.h(-1).kg(-1) (coefficient of variation [CV], 51%), volume of distribution was 2.12 l/kg (CV, 34%), and absorption rate constant was 1.61 h(-1) (CV, 46%). Mean absorption and elimination half-lives were 0.43 and 5.1 h, respectively. Age and percentage of free ropivacaine were significant covariates for clearance. Posterior Bayesian estimates of clearance were significantly higher (38%) in older children. CONCLUSION: Total and free plasma ropivacaine concentrations after caudal ropivacaine (0.2%, 2 mg/kg) in infants were within the range of concentrations previously reported in adults and older children. Age and percentage of free ropivacaine were significant covariates of clearance. PMID- 11379677 TI - Automated responsiveness test (ART) predicts loss of consciousness and adverse physiologic responses during propofol conscious sedation. AB - BACKGROUND: The authors evaluated a device designed to provide conscious sedation with propofol (propofol-air), or propofol combined with 50% nitrous oxide (N2O; propofol-N2O). An element of this device is the automated responsiveness test (ART), a method for confirming that patients remain conscious. The authors tested the hypotheses that the ART predicts loss of consciousness and that failure to respond to the ART precedes sedation-induced respiratory or hemodynamic toxicity. METHODS: The protocol consisted of sequential 15-min cycles in 20 volunteers. After a 15-min control period, propofol was infused to an initial target effect site concentration of 0.0 microg/ml with N2O or 1.5 microg/ml with air. Subsequently, the propofol target effect-site concentration was increased by a designated increment (0.25 and 0.5 microg/ml) and the process repeated. This sequence was continued until loss of consciousness, as defined by an Observer's Assessment of Alertness/Sedation (OAA/S) score of 10/20 or less, or until an adverse physiologic event was detected. RESULTS: The OAA/S score at which only 50% of the volunteers were able to respond to the ART (P50) during propofol-N2O was 11.1 of 20 (95% confidence interval [CI]: 10.6-11.8); the analogous P50 was 11.8 of 20 (95% CI: 11.4-12.3) with propofol-air. Failure to respond to the ART occurred at a plasma propofol concentration of 0.7 +/- 0.6 microg/ml with propofol-N2O and 1.6 +/- 0.6 microg/ml with propofol-air, whereas loss of consciousness occurred at 1.2 +/- 0.8 microg/ml and 1.9 +/- 0.7 microg/ml, respectively. There were no false-normal ART responses. CONCLUSION: The ART can guide individual titration of propofol because failure to respond to responsiveness testing precedes loss of consciousness and is not susceptible to false-normal responses. The use of N2O with propofol for conscious sedation decreases the predictive accuracy of the ART. PMID- 11379678 TI - Minimum local analgesic dose of intrathecal bupivacaine in labor and the effect of intrathecal fentanyl. AB - BACKGROUND: Combining bupivacaine with fentanyl for intrathecal analgesia in labor is well recognized, but dosages commonly used are arbitrarily chosen and may be excessive. This study aimed to determine the median effective dose (ED50) of intrathecal bupivacaine, defined as the minimum local analgesic dose (MLAD), and then use this to assess the effect of different doses of fentanyl. METHODS: In this double-blind, randomized, prospective study, 124 parturients receiving combined spinal epidural analgesia at 2-6-cm cervical dilatation were allocated to one of four groups to receive bupivacaine alone or with 5, 15, or 25 microg fentanyl, using the technique of up-down sequential allocation. Analgesic effectiveness was assessed using 100-mm visual analog pain scores, with less than or equal to 10 mm within 15 min defined as effective. MLAD was calculated using the formula of Dixon and Massey. Pruritus and duration of spinal analgesia were also recorded. RESULTS: Minimum local analgesic dose of intrathecal bupivacaine was 1.99 mg (95% confidence interval, 1.71, 2.27). There were similar significant reductions in MLAD (P < 0.001) for all bupivacaine-fentanyl groups compared with bupivacaine control. There was a dose-dependent increase in both pruritus and duration of spinal analgesia with increasing fentanyl (P < 0.0001). CONCLUSION: Under the conditions of this study, the addition of intrathecal fentanyl 5 microg offers a similar significant bupivacaine dose-sparing effect as 15 and 25 microg. Analgesia in the first stage of labor can be achieved using lower doses of fentanyl, resulting in less pruritus but with a shortening of duration of action. PMID- 11379679 TI - Preemptive analgesic effects of ketorolac in ankle fracture surgery. AB - BACKGROUND: Preemptive analgesia has been difficult to show in human experiments. If ketorolac has preemptive effects, then there may be an advantage to administering it at the beginning of surgery despite the potential for increased blood loss. METHODS: The authors performed a randomized, double-blind, controlled trial of 48 patients scheduled for ankle fracture surgery in a county trauma hospital. Anesthesia management was standardized and included adequate opioid analgesia (5 microg/kg fentanyl and 0.1 mg/kg morphine). Intravenous 30 mg ketorolac was administered to 23 patients before tourniquet inflation and to 25 patients after tourniquet inflation. Visual analog scale pain scores, morphine patient-controlled analgesia consumption, nausea-vomiting, and postoperative bleeding were measured. RESULTS: The 23 patients given ketorolac before tourniquet inflation had no increase in pain postoperatively compared with their preoperative baseline (P = 0.280). The 25 patients who received ketorolac minutes later after tourniquet inflation had significant increases in their postoperative pain compared with their preoperative baseline (P = 0.00116). This effect was short-lived, and by 6 h the pain score in this group was not significantly more than it was preoperatively. Intergroup comparison showed a lower visual analog scale score at 2 (P = 0.0203) and 4 h (P = 0.00549) in the preemptive group and lower nausea scores at hour 6 (P = 0.00704). There was no difference in patient controlled analgesia consumption between groups. CONCLUSIONS: Intravenous 30 mg ketorolac appears to have preemptive analgesic effects in patients undergoing ankle fracture repair. Ketorolac administered before tourniquet inflation prevents postoperative pain being perceived as more intense than preoperative pain. PMID- 11379680 TI - The effects of CO2 on respiratory mechanics in anesthetized paralyzed humans. AB - BACKGROUND: There is little information concerning the carbon dioxide-related effects on respiratory mechanics in anesthetized, paralyzed subjects; however, hypocapnia or hypercapnia is often permitted in patients with severe brain injury or acute respiratory distress syndrome. Therefore, the carbon dioxide dependence of respiratory mechanics in healthy anesthetized, paralyzed subjects was investigated. METHODS: Interrupter resistance (Rint), additional tissue viscoelastic resistance (deltaR), and quasi-static elastance (Est) of lung (L) and chest wall were assessed by means of the rapid end-inspiratory occlusion method in two groups of seven healthy paralyzed subjects anesthetized with diazepam or isoflurane. They underwent ventilation with a fixed pattern and hyperoxic gas mixtures with different fractions of inspired carbon dioxide (FICO2) to produce a partial pressures of arterial carbon dioxide (PaCO2) of 24.4 +/- 3.4, 39.6 +/- 3.2, and 62 +/- 4.1 (SD) mmHg. RESULTS: Chest wall mechanics and Est,L were unaffected by PaCO2 changes. With diazepam anesthesia, Rint,L decreased linearly, with increasing PaCO2, from 2.3 to 1.4 cm H2O.s.l(-1), whereas deltaR,L decreased from 2 to 1.7 cm H2O.s.l(-1), though not significantly. With isoflurane anesthesia, the decrease of Rint,L (0.2 +/- 0.5 cm H2O.s.l(-1)) was not significant, and deltaRL remained unchanged. With diazepam, Rint,L was 45 (hypercapnia) to 110% (hypocapnia) greater than with isoflurane. CONCLUSIONS: Changes of PaCO2 from 20-65 mmHg cause increasing bronchodilation in anesthetized, paralyzed subjects, this effect being attenuated or abolished by drugs (e.g., halogenated anesthetics) that depress smooth muscle tone substantially. The carbon dioxide bronchodilating effects are probably direct for peripheral structures and are paralleled by a tendency of lung tissue resistance to decrease. Because local PaCO2-related changes in bronchomotor tone promote VA/Q matching, this mechanism should be impaired by anesthetics that cause bronchodilation. PMID- 11379681 TI - Minimum alveolar concentration (MAC) of xenon with sevoflurane in humans. AB - BACKGROUND: Although more than 30 yr ago the minimum alveolar concentration (MAC) of xenon was determined to be 71%, that previous study had technological limitations, and no other studies have confirmed the MAC value of xenon since. The current study was designed to confirm the MAC value of xenon in adult surgical patients using more modern techniques. METHODS: Sixty patients were anesthetized with sevoflurane with or without xenon. They were randomly allocated to one of four groups; patients in group 1 received no xenon, whereas those in groups 2, 3, and 4 received end-tidal concentrations of 20, 40, and 60%, respectively (n = 15 each group). Target end-tidal sevoflurane concentrations were chosen using the "up-and-down" method in each group. After steady state sevoflurane and xenon concentrations were maintained for at least 15 min, each patient was monitored for a somatic response at surgical incision. Somatic response was defined as any purposeful bodily movement. The MAC of sevoflurane and its reduction by xenon was evaluated using the multiple independent variable logistic regression model. RESULTS: The interaction coefficient of the multiple variable logistic regression was not significantly different from zero (P = 0.143). The MAC of xenon calculated as xenon concentration that would reduce MAC of sevoflurane to 0% was 63.1%. CONCLUSIONS: The authors could not determine whether interaction in blocking somatic responses in 50% of patients is additive. The MAC of xenon is in the range of the values that were predicted in a previous study. PMID- 11379682 TI - Comparison of multiplane transesophageal echocardiography and contrast-enhanced helical CT in the diagnosis of blunt traumatic cardiovascular injuries. AB - BACKGROUND: Multiplane transesophageal echocardiography (TEE) and helical computed tomography (CT) of the chest have been validated separately against aortography for the diagnosis of acute traumatic aortic injuries (ATAI). However, their respective diagnostic accuracy in identifying blunt traumatic cardiovascular lesions has not been compared. METHODS: During a 3-yr period, 110 consecutive patients with severe blunt chest trauma (age: 41 +/- 17 yr; injury severity score: 34 +/- 14) prospectively underwent TEE and chest CT as part of their initial evaluation. Results of both imaging methods were interpreted independently by experienced investigators and subsequently compared. All cases of subadventitial acute traumatic aortic injury were surgically confirmed. RESULTS: Seventeen patients had vascular injury and 11 had cardiac lesions. TEE and CT identified all subadventitial disruptions involving the aortic isthmus (n = 10) or the ascending aorta (n = 1) that necessitated surgical repair. In contrast, CT only depicted one disruption of the innominate artery. TEE detected injuries involving the intimal or medial layer, or both, of the aortic isthmus in four patients with apparently normal CT results who underwent successful conservative treatment. All cardiac injuries but two were identified only by TEE. CONCLUSIONS: In patients with severe blunt chest trauma, TEE and CT have similar diagnostic accuracy for the identification of surgical acute traumatic aortic injuy. TEE also allows the diagnosis of associated cardiac injuries and is more sensitive than CT for the identification of intimal or medial lesions of the thoracic aorta. PMID- 11379683 TI - Effects of ketamine and its isomers on ischemic preconditioning in the isolated rat heart. AB - BACKGROUND: Ischemic preconditioning protects the heart against subsequent ischemia. Opening of the adenosine triphosphate-sensitive potassium (KATP) channel is a key mechanism of preconditioning. Ketamine blocks KATP channels of isolated cardiomyocytes. The authors investigated the effects of ketamine and its stereoisomers on preconditioning. METHODS: Isolated rat hearts (n = 80) underwent 30 min of no-flow ischemia and 60 min of reperfusion. Two groups with eight hearts each underwent the protocol without intervention (control-1 and control 2), and, in eight hearts, preconditioning was elicited by two 5-min periods of ischemia before the 30 min ischemia. In the six treatment groups (each n = 8), ketamine, R(-)- or S(+)-ketamine were administered at concentrations of 2 or 20 microg/ml before preconditioning. Eight hearts received 20 microg/ml R(-) ketamine before ischemia. Left ventricular (LV) developed pressure and creatine kinase (CK) release during reperfusion were determined as variables of ventricular function and cellular injury. RESULTS: Baseline LV developed pressure was similar in all groups: 104 +/- 28 mmHg (mean +/- SD). Controls showed a poor recovery of LV developed pressure (17 +/- 8% of baseline) and a high CK release (70 +/- 17 IU/g). Ischemic preconditioning improved recovery of LV developed pressure (46 +/- 14%) and reduced CK release (47 +/- 17 IU/g, both P < 0.05 vs. control-1). Ketamine (2 microg/ml) and 2 or 20 microg/ml S(+)-ketamine had no influence on recovery of LV developed pressure compared with preconditioning (47 +/- 18, 43 +/- 8, 49 +/- 36%) and CK release (39 +/- 8, 30 +/- 14, 41 +/- 25 IU/g). After administration of 20 microg/ml ketamine and 2 or 20 microg/ml R(-) ketamine, the protective effects of preconditioning were abolished (LV developed pressure-recovery, 16 +/- 14, 22 +/- 21, 18 +/- 11%; CK release, 67 +/- 11, 80 +/ 21, 82 +/- 41 IU/g; each P < 0.05 vs. preconditioning). Preischemic treatment with R(-)-ketamine had no effect on CK release (74 +/- 8 vs. 69 +/- 9 IU/g in control-2, P = 0.6) and functional recovery (LV developed pressure 12 +/- 4 vs. 9 +/- 2 mmHg in control-2, P = 0.5). CONCLUSION: Ketamine can block the cardioprotective effects of ischemic preconditioning. This effect is caused by the R(-)-isomer. PMID- 11379684 TI - Ketamine, but not S(+)-ketamine, blocks ischemic preconditioning in rabbit hearts in vivo. AB - BACKGROUND: Ketamine blocks KATP channels in isolated cells and abolishes the cardioprotective effect of ischemic preconditioning in vitro. The authors investigated the effects of ketamine and S(+)-ketamine on ischemic preconditioning in the rabbit heart in vivo. METHODS: In 46 alpha-chloralose anesthetized rabbits, left ventricular pressure (tip manometer), cardiac output (ultrasonic flow probe), and myocardial infarct size (triphenyltetrazolium staining) at the end of the experiment were measured. All rabbits were subjected to 30 min of occlusion of a major coronary artery and 2 h of subsequent reperfusion. The control group underwent the ischemia-reperfusion program without preconditioning. Ischemic preconditioning was elicited by 5-min coronary artery occlusion followed by 10 min of reperfusion before the 30 min period of myocardial ischemia (preconditioning group). To test whether ketamine or S(+) ketamine blocks the preconditioning-induced cardioprotection, each (10 mg kg(-1)) was administered 5 min before the preconditioning ischemia. To test any effect of ketamine itself, ketamine was also administered without preconditioning at the corresponding time point. RESULTS: Hemodynamic baseline values were not significantly different between groups [left ventricular pressure, 107 +/- 13 mmHg (mean +/- SD); cardiac output, 183 +/- 28 ml/min]. During coronary artery occlusion, left ventricular pressure was reduced to 83 +/- 14% of baseline and cardiac output to 84 +/- 19%. After 2 h of reperfusion, functional recovery was not significantly different among groups (left ventricular pressure, 77 +/- 19%; cardiac output, 86 +/- 18%). Infarct size was reduced from 45 +/- 16% of the area at risk in controls to 24 +/- 17% in the preconditioning group (P = 0.03). The administration of ketamine had no effect on infarct size in animals without preconditioning (48 +/- 18%), but abolished the cardioprotective effects of ischemic preconditioning (45 +/- 19%, P = 0.03). S(+)-ketamine did not affect ischemic preconditioning (25 +/- 11%, P = 1.0). CONCLUSIONS: Ketamine, but not S(+)-ketamine blocks the cardioprotective effect of ischemic preconditioning in vivo. PMID- 11379685 TI - Effect of increasing perfluorocarbon dose on VA/Q distribution during partial liquid ventilation in acute lung injury. AB - BACKGROUND: Although gas exchange during partial liquid ventilation (PLV) depends on perfluorocarbon liquid, the effect of perfluorocarbon dose on the ventilation perfusion (VA/Q) distribution is not known. This study investigated how VA/Q distribution of an acutely injured lung is affected during PLV at increasing perfluorocarbon dose. METHODS: In eight rabbits (3.2 +/- 0.1 kg), acute lung injury (ALI) was created by repeated saline lavage (arterial oxygen partial pressure/fraction of inspired oxygen, 37 +/- 11 mm Hg). Three different doses of perfluorodecalin (9 ml/kg = low dose; 13.5 ml/kg = medium dose; 18 ml/kg = functional residual capacity [FRC] dose) were applied in random order during PLV. VA/Q distribution at different doses was evaluated by multiple inert gas elimination technique. RESULTS: Inert gas shunt (63 +/- 21% at ALI) decreased with increasing perfluorocarbon dose (43 +/- 21% at low dose, 29 +/- 10% at medium dose, 11 +/- 9% at FRC dose; P = 0.022). Compared with ALI (0%), the proportion of low VA/Q units was higher at all tested doses (19 +/- 10, 25 +/- 12, and 34 +/- 18%, respectively; all P < 0.05). Compared with ALI (27 +/- 14%), the proportion of normal VA/Q units was not increased at low or medium doses but was increased only at the FRC dose (45 +/- 13%; P = 0.027). CONCLUSIONS: With increasing perfluorocarbon dose during PLV, shunt was reduced from a small dose. The majority shunt units were converted to units showing low VA/Q ratios rather than normal VA/Q ratios. The presence of considerable amount of low VA/Q units across the varying doses of perfluorocarbon suggested that additional measures are necessary during PLV to augment its effect on gas exchange. PMID- 11379686 TI - Blockade and activation of the human neuronal nicotinic acetylcholine receptors by atracurium and laudanosine. AB - BACKGROUND: Curaremimetic nondepolarizing muscle relaxants are widely used in clinical practice to prevent muscle contraction either during surgery or during intensive care. Although primarily acting at the neuromuscular junction, these compounds can cause adverse effects, including modification of cardiac rhythm, arterial blood pressure, and in the worst cases, triggering of seizures. In this study, we assessed the interaction of atracurium and its metabolite, laudanosine, with neuronal nicotinic receptors. METHODS: The human neuronal nicotinic receptors alpha4beta2, alpha3beta4, alpha3alpha5beta4, and alpha7 are heterologously expressed in Xenopus laevis oocytes, and the effect of atracurium and its degradation product, laudanosine, were studied on these receptors. RESULTS: Atracurium and laudanosine inhibited in the micromolar range the major brain alpha4beta2 receptor and the ganglionic alpha3beta4 or alpha3beta4alpha5 and the homomeric alpha7 receptors. For all four receptors, inhibition was rapid and readily reversible within less than 1 min. Atracurium blockade was competitive at alpha4beta2 and alpha7 receptors but displayed a noncompetitive blockade at the alpha3beta4 receptors. Inhibition at this receptor subtype was not modified by alpha5. Laudanosine was found to have a dual mode of action; first, it competes with acetylcholine and, second, it blocks the ionic pore by steric hindrance. At low concentrations, these two drugs are able to activate both the alpha4beta2 and the alpha3beta4 receptors. CONCLUSION: Adverse effects observed during atracurium administration may be attributed, at least partly, to an interaction with neuronal nicotinic receptors. PMID- 11379687 TI - Propofol attenuates diaphragmatic dysfunction induced by septic peritonitis in hamsters. AB - BACKGROUND: Sepsis or peritonitis impairs diaphragmatic contractility and endurance capacity. Peroxynitrite, a powerful oxidant formed by superoxide and nitric oxide, has been implicated in the pathogenesis. Propofol scavenges this reactive molecule. The authors conducted the current study to evaluate whether propofol prevents diaphragmatic dysfunction induced by septic peritonitis. METHODS: Forty male Golden-Syrian hamsters (120-140 g) were randomly classified into five groups. Groups sham and sham-propofol 50 underwent sham laparotomy alone, whereas groups sepsis, sepsis-propofol 25, and sepsis-propofol 50 underwent cecal ligation with puncture. Groups sham and sepsis received infusion of intralipid, whereas groups sham-propofol 50, sepsis-propofol 25, and sepsis propofol 50 received propofol at rates of 50, 25, and 50 mg.kg(-1).h(-1), respectively. Intralipid or propofol was subcutaneously infused from 3 h before surgery until 24 h after operation, when all hamsters were killed. Diaphragmatic contractility and fatigability were assessed in vitro using diaphragm muscle strips. Peroxynitrite formation in the diaphragm was assessed by nitrotyrosine immunostaining. Plasma nitrite-nitrate concentrations and diaphragmatic concentrations of malondialdehyde were determined. Using another set of animals, diaphragmatic inducible nitric oxide synthase activity was also measured. RESULTS: Twitch, tetanic tensions, and tensions during fatigue trials were reduced in group sepsis compared with group sham. In group SEPSIS, diaphragm malondialdehyde and inducible nitric oxide synthase activity, and plasma nitrite nitrate concentrations increased, and positive immunostaining for nitrotyrosine residues was found. Propofol attenuated these changes. CONCLUSIONS: Pretreatment with propofol attenuated diaphragmatic dysfunction induced by septic peritonitis in hamsters assessed by contractile profiles and endurance capacity. This beneficial effect of propofol may be caused, in part, by inhibition of lipid peroxidation in the diaphragm caused by the powerful oxidant. PMID- 11379688 TI - Amitriptyline versus bupivacaine in rat sciatic nerve blockade. AB - BACKGROUND: Amitriptyline, a tricyclic antidepressant, is frequently used orally for the management of chronic pain. To date there is no report of amitriptyline producing peripheral nerve blockade. The authors therefore investigated the local anesthetic properties of amitriptyline in rats and in vitro. METHODS: Sciatic nerve blockade was performed with 0.2 ml amitriptyline or bupivacaine at selected concentrations, and the motor, proprioceptive, and nociceptive blockade was evaluated. Cultured rat GH3 cells were externally perfused with amitriptyline or bupivacaine, and the drug affinity toward inactivated and resting Na+ channels was assessed under whole-cell voltage clamp conditions. In addition, use dependent blockade of these drugs at 5 Hz was evaluated. RESULTS: Complete sciatic nerve blockade for nociception was obtained with amitriptyline for 217 +/ 19 min (5 mM, n = 8, mean +/- SEM) and for 454 +/- 38 min (10 mM, n = 7) versus bupivacaine for 90 +/- 13 min (15.4 mM, n = 6). The time to full recovery of nociception for amitriptyline was 353 +/- 12 min (5 mM) and 656 +/- 27 min (10 mM) versus 155 +/- 9 min for bupivacaine (15.4 mM). Amitriptyline was approximately 4.7-10.6 times more potent than bupivacaine in binding to the resting channels (50% inhibitory concentration [IC50] of 39.8 +/- 2.7 vs. 189.6 +/- 22.3 microM) at - 150 mV, and to the inactivated Na+ channels (IC50 of 0.9 +/ 0.1 vs. 9.6 +/- 0.9 microM) at -60 mV. High-frequency stimulation at 3 microM caused an additional approximately 14% blockade for bupivacaine, but approximately 50% for amitriptyline. CONCLUSION: Amitriptyline is a more potent blocker of neuronal Na+ channels than bupivacaine in vivo and in vitro. These findings suggest that amitriptyline could extend its clinical usefulness for peripheral nerve blockade. PMID- 11379690 TI - Cardiovascular responses to the induction of mild hypothermia in the presence of epidural anesthesia. AB - BACKGROUND: The combining of epidural anesthesia with general anesthesia impairs central and peripheral thermoregulatory control and therefore is often accompanied by unintended intraoperative hypothermia. However, little is known about the cardiovascular response to hypothermia during combined epidural and general anesthesia. The authors assessed the effects of hypothermia during such combined anesthesia. METHODS: The authors randomly assigned 30 mongrel dogs anesthetized with isoflurane (1.0%) to three groups of 10: control, receiving general anesthesia alone; thoracic injection, additionally receiving thoracic epidural anesthesia; and lumbar injection, additionally receiving thoracolumbar epidural anesthesia. Core temperature was lowered from 38.5 degrees C to approximately 34 degrees C (mild hypothermia) using a femoral arteriovenous shunt in an external cool water bath. During hypothermia, the authors measured heart rate, cardiac output, and plasma catecholamine concentrations in each group. Ejection fraction was also measured using echocardiography. RESULTS: Compared with measurements during baseline conditions (general anesthesia alone with no epidural injection and no hypothermia) in the control, thoracic, and lumbar injection groups, the injections followed by hypothermia produced 17, 32, and 41% decreases in heart rate; 22, 32, and 47% reductions in cardiac output; 66, 85, and 92% decreases in the epinephrine concentrations; and 27, 44, and 85% decreases in the norepinephrine concentrations. In contrast, ejection fraction did not change in any group. CONCLUSION: Mild hypothermia during combined epidural anesthesia and general anesthesia markedly reduced cardiac output in dogs, mainly by decreasing heart rate. PMID- 11379689 TI - Inhibitory effects of etomidate and ketamine on endothelium-dependent relaxation in canine pulmonary artery. AB - BACKGROUND: The authors recently demonstrated that acetylcholine-induced pulmonary vasorelaxation had two primary components, nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF). The goal was to investigate the effects of etomidate and ketamine on the NO- and EDHF-mediated components of pulmonary vasorelaxation in response to acetylcholine, bradykinin, and the calcium ionophore, A23187. METHODS: Canine pulmonary arterial rings with an intact endothelium were suspended in organ chambers for isometric tension recording. The effects of etomidate and ketamine (10(-5) M and 10(-4) M) on vasorelaxation responses to acetylcholine, bradykinin, and A23187 were assessed in phenylephrine-contracted rings. The NO- and EDHF-mediated components of relaxation were assessed using a NO synthase inhibitor (N-nitro-L-arginine methylester [L-NAME]: 10(-4) M) and a Ca2+-activated potassium channel inhibitor (tetrabutylammonium hydrogen sulfate [TBA]: 10(-3) M) in rings pretreated with a cyclooxygenase inhibitor (ibuprofen: 10(-5) M). Intracellular calcium concentration ([Ca2+]i) was measured in cultured bovine pulmonary artery endothelial cells loaded with acetoxylmethyl ester of fura-2. RESULTS: Etomidate and ketamine attenuated pulmonary vasorelaxation in response to acetylcholine and bradykinin, whereas they had no effect on the response to A23187. The relaxant responses to acetylcholine and bradykinin were attenuated by L-NAME or TBA alone and were abolished by combined inhibition in rings pretreated with ibuprofen. Etomidate and ketamine further attenuated both L-NAME-resistant and TBA-resistant relaxation. These anesthetics also inhibited increases in endothelial [Ca2+]i in response to bradykinin, but not A23187. CONCLUSION: These results indicate that etomidate and ketamine attenuated vasorelaxant responses to acetylcholine and bradykinin by inhibiting both NO- and EDHF-mediated components. Moreover, our results suggest that these anesthetics do not directly suppress NO or EDHF activity, but rather inhibit the endothelial [Ca2+]i transient in response to receptor activation. PMID- 11379691 TI - Different inhibitory effects of volatile anesthetics on T- and L-type voltage dependent Ca2+ channels in porcine tracheal and bronchial smooth muscles. AB - BACKGROUND: The distal airway is more important in the regulation of airflow resistance than is the proximal airway, and volatile anesthetics have a greater inhibitory effect on distal airway muscle tone. The authors investigated the different reactivities of airway smooth muscles to volatile anesthetics by measuring porcine tracheal or bronchial (third to fifth generation) smooth muscle tension and intracellular concentration of free Ca2+ ([Ca2+]i) and by measuring inward Ca2+ currents (ICa) through voltage-dependent Ca2+ channels (VDCs). METHODS: Intracellular concentration of free Ca2+ was monitored by the 500-nm light emission ratio of Ca2+ indicator fura-2. Isometric tension was measured simultaneously. Whole-cell patch clamp recording techniques were used to investigate the effects of volatile anesthetics on ICa in dispersed smooth muscle cells. Isoflurane (0-1.5 minimum alveolar concentration) or sevoflurane (0-1.5 minimum alveolar concentration) was introduced into a bath solution. RESULTS: The volatile anesthetics tested had greater inhibitory effects on carbachol-induced bronchial smooth muscle contraction than on tracheal smooth muscle contraction. These inhibitory effects by the anesthetics on muscle tension were parallel to the inhibitory effects on [Ca2+]i. Although tracheal smooth muscle cells had only L-type VDCs, some bronchial smooth muscle cells (approximately 30%) included T type VDC. Each of the two anesthetics significantly inhibited the activities of both types of VDCs in a dose-dependent manner; however, the anesthetics had greater inhibitory effects on T-type VDC activity in bronchial smooth muscle. CONCLUSIONS: The existence of the T-type VDC in bronchial smooth muscle and the high sensitivity of this channel to volatile anesthetics seem to be, at least in part, responsible for the different reactivities to the anesthetics in tracheal and bronchial smooth muscles. PMID- 11379692 TI - Inhibitory effects of barbiturates on nicotinic acetylcholine receptors in rat central nervous system neurons. AB - BACKGROUND: Neuronal nicotinic acetylcholine receptors (nAChRs) are widely expressed in the central and autonomic nervous systems. The authors have previously shown that depressant and convulsant barbiturates both inhibit the ganglion-type nAchRs in PC12 cells. However, the central and gangliontype receptors have different subunit composition and pharmacologic properties. In this study, the authors investigated the effects of thiopental, depressant [R(-)] and convulsant [S(+)] stereoisomers of 1-methyl-5 phenyl-5-propyl barbituric acid (MPPB) on neuronal nAChRs in the rat central nervous system to explore significance of these effects in barbiturate anesthesia. METHODS: Whole-cell currents were measured in acutely dissociated rat medial habenula (MHb) neurons by applying 10 or 100 microM nicotine in the absence or presence of thiopental 3 100 microM. Effects of R(-)- and S(+)-MPPB on the nicotine-induced current were also studied. RESULTS: Thiopental suppressed the nicotine-elicited inward current and accelerated the current decay dose-dependently at the clinical relevant concentrations. R(-)- and S(+)-MPPB both inhibited the nicotine-induced current dose-dependently without augmenting the current decay. There was no significant difference in the magnitudes of inhibition by R(-)- and S(+)-MPPB. CONCLUSIONS: Although thiopental suppressed the current mediated through native nAchRs in rat MHb neurons at the clinically relevant concentrations, the depressant and convulsant stereoisomers of MPPB both inhibited the current in the same extent. These findings are consistent with the results previously obtained in the ganglion-type receptors of PC12 cells and suggest that inhibition of nAChRs in MHb neurons is not directly relevant to the hypnotic or anticonvulsive actions of barbiturates. PMID- 11379693 TI - Compartment syndrome in surgical patients. PMID- 11379694 TI - The use of intravenous nitroglycerin in a case of spasm of the sphincter of Oddi during laparoscopic cholecystectomy. AB - Spasm of the sphincter of Oddi still occurs during cholecystectomy. Some reports indicate that the spasm, induced by morphine, can be reversed by injection of naloxone, nalbuphine, and glucagon. Others maintain that nitroglycerin or nifedipine can relax the sphincter of Oddi muscle. We recently encountered spasm of the sphincter of Oddi during a laparoscopic cholecystectomy and treated it successfully with intravenous nitroglycerin. PMID- 11379695 TI - Electrocautery-induced tachycardia in a rate-responsive pacemaker. AB - Electric interference by cautery on demand pacemakers can cause inhibition and loss of pacing. We report a case in which electrocautery induced a pacemaker in ventricle-paced, ventricle-sensed, inhibited, rate-responsive (VVIR) mode to pace at a programmed maximum rate of 130 pulses/min. PMID- 11379696 TI - Cardiac surgery increases surgical complexity. PMID- 11379697 TI - Patient outcomes and "directed" anesthesia care. PMID- 11379698 TI - Measuring the influence of anesthesiologists' medical direction. PMID- 11379699 TI - Medical direction during anesthesia: what or who is the problem? PMID- 11379700 TI - Abnormal serum sodium and chloride concentrations may contribute to cognitive dysfunction during severe isovolemic anemia. PMID- 11379701 TI - The effect of lumbar plexus block on blood loss and postoperative pain. PMID- 11379702 TI - Rapid extraction of middle-latency auditory-evoked potentials. PMID- 11379703 TI - Tracheostomy tube replacement: role of the airway exchange catheter. PMID- 11379704 TI - Transient lingual and glossopharyngeal nerve injury: a complication of cuffed oropharyngeal airway. PMID- 11379705 TI - Prevalence of ceftriaxone- and ceftazidime-resistant gram-negative bacteria in long-term-care facilities. PMID- 11379706 TI - In vitro activity of a nonmedicated handwash product, chlorhexidine, and an alcohol-based hand disinfectant against multiply resistant gram-positive microorganisms. PMID- 11379707 TI - High methicillin resistance of Staphylococcus aureus and coagulase-negative staphylococci in Imam Khomeini Hospital of Urmia, Iran. PMID- 11379708 TI - Why don't doctors and nurses wash their hands? PMID- 11379709 TI - Bacterial contamination associated with electronic faucets: a new risk for healthcare facilities. AB - OBJECTIVE: To investigate the safety of the hospital water supply following a major flood. DESIGN: Surveillance was conducted of the hospital water supply as it entered the hospital and at randomly selected water faucets throughout the facility. SETTING: A newly constructed surgical critical-care unit in a 265-bed community hospital that had to be evacuated and was out of operation for 6 weeks following a major flood of the city. METHODS: Random water samples throughout the facility were analyzed for heterotrophic plate counts (HPCs), chlorine, and coliforms utilizing standard methods. RESULTS: Water samples entering the hospital met appropriate standards, indicating the city water distribution system was not contaminated. Of 169 faucets tested, 13 (22%) of 59 electronic faucets exceeded the HPC threshold, and 12 (11%) of 110 manual faucets exceeded the HPC threshold (P<.14). A comparison of two brands of electronic faucets with manual faucets and with each other revealed that the HPC threshold was exceeded by 11 (32%) of 34 brand A faucets as compared to 12 (11%) of 110 manual faucets (P<.006). The HPC threshold was exceeded by 2 (8%) of 25 brand B faucets compared to 12 (11%) of 110 manual faucets (P<.94). Contamination rates of brand A and brand B faucets differed significantly (P<.003). Similar testing 2 months after hyperchlorination of the water supply indicated that the HPC threshold was exceeded by 16 (52%) of 31 brand A faucets compared to 10 (9.%) of 110 manual faucets (P<.0000003) and by 2 (18%) of 25 brand B faucets compared to 10 (9%) of 110 manual faucets (P=1.0). CONCLUSIONS: A certain brand of electronic water faucet used in the hospital was associated with unacceptable levels of microbial growth in water and was a continuing source of bacteria potentially hazardous to patients. PMID- 11379710 TI - Risk of exposure to bloodborne infection for Italian healthcare workers, by job category and work area. Studio Italiano Rischio Occupazionale da HIV Group. AB - OBJECTIVE: To analyze the rate of occupational exposure to blood and body fluids from all sources and specifically from human immunodeficiency virus (HIV) infected sources among hospital workers, by job category and work area. DESIGN: Multicenter prospective study. Occupational exposure data (numerator) and full time equivalents ([FTEs] denominator) were collected over a 5-year period (1994 1998) and analyzed. SETTING: 18 Italian urban acute-care hospitals with infectious disease units. RESULTS: A total of 10,988 percutaneous and 3,361 mucocutaneous exposures were reported. The highest rate of percutaneous exposure per 100 FTEs was observed among general surgery (11%) and general medicine (10.6%) nurses, the lowest among infectious diseases (1.1%) and laboratory (1%) physicians. The highest rates of mucocutaneous exposure were observed among midwives (5.3%) and dialysis nurses (4.7%), the lowest among pathologists (0%). Inadequate sharps disposal and the prevalence of sharps in the working unit influence the risk to housekeepers. The highest combined HIV exposure rates were observed among nurses (7.8%) and physicians (1.9%) working in infectious disease units. The highest rates of high-risk percutaneous exposures per 100 FTE were again observed in nurses regardless of work area, but this risk was higher in medical areas than in surgery (odds ratio, 2.1; 95% confidence interval, 1.9-2.5; P<.0001). CONCLUSION: Exposure risk is related to job tasks, as well as to the type and complexity of care provided in different areas, whereas HIV exposure risk mainly relates to the prevalence of HIV-infected patients in a specific area. The number of accident-prone procedures, especially those involving the use of hollow-bore needles, performed by job category influence the rate of exposure with high risk of infection. Job- and area-specific exposure rates permit monitoring of the effectiveness of targeted interventions and control measures over time. PMID- 11379711 TI - Methicillin-resistant Staphylococcus aureus in two tertiary-care centers in Jeddah, Saudi Arabia. AB - OBJECTIVE: To review clinical experience with methicillin-resistant Staphylococcus aureus (MRSA) in tertiary-care hospitals in Jeddah, Saudi Arabia. DESIGN: Retrospective review for the year 1998. SETTING: Two tertiary-care hospitals. METHODS: Results of MRSA-positive cultures of clinical specimens obtained as part of investigations for suspected infections were retrieved from the microbiology laboratories' records. Charts of patients were reviewed, with standardized data collection. RESULTS: Of 673 S. aureus isolates identified, 222 (33%, or 6.8 isolates/1,000 admissions) were MRSA. Overall MRSA prevalence was 2% in 1988. Nosocomial acquisition occurred in 84.2% of cases. All age groups were affected, and 52% of patients had at least one comorbidity. MRSA prevalence was highest in the intensive care units (26.6% of all isolates), the medical wards (24.8%), and the surgical wards (19.8%). Seventy-three percent of isolates caused infection; the rest represented colonization. Surgical wounds (35.2%), the chest (29%), and central venous catheters (13%) were the most common sites of infection. Bacteremia occurred in 15.4% of patients. Local signs (84%) and fever (75.9%) were the most common clinical manifestations. Respiratory distress and septic shock occurred in 30.2% and 13.6% of cases, respectively. Of 162 patients with MRSA infection and 60 patients with MRSA colonization, 95.7% and 70% received antibiotics in the preceding 6 weeks, respectively (P<.0001). The total mortality of patients with MRSA infection was 53.7%: 36.4% as a result of MRSA infection and 17.3% as a result of other causes. CONCLUSIONS: The prevalence of MRSA is high and rapidly increasing in the two hospitals, as it is worldwide. Control measures to prevent the spread of MRSA in hospitals should continue, with reinforcement of hygienic precautions and development of policies to restrict the use of antibiotics. PMID- 11379712 TI - Management of an outbreak of vancomycin-resistant enterococci in the medical intensive care unit of a cancer center. AB - Between November 1996 and February 1997, 17 episodes of vancomycin-resistant enterococci (VRE) infection or colonization (9 infections, 8 colonizations), all with the same or a similar genomic DNA pattern, were identified in the medical intensive care unit (MICU) of a tertiary-care cancer hospital. The cases were genotypically traced to a patient who was admitted to the hospital in September 1996 and who, by December 1996, had four different admissions to the MICU. Multifaceted infection control measures, including decontamination of the environment and of nondisposable equipment, halted the nosocomial transmission of VRE in the MICU. PMID- 11379713 TI - A comparison of multifaceted versus Clostridium difficile-focused VRE surveillance strategies in a low-prevalence setting. AB - We compared our current screening strategy for vancomycin-resistant Enterococcus (VRE) with a focused strategy that screens all stool samples sent for Clostridium difficile toxin assay but limits rectal swab screening to wards with new VRE cases detected via C. difficile samples. The proposed strategy detects 72.7% of new VRE cases, with substantial cost savings. PMID- 11379714 TI - Guidelines for the management of intravascular catheter-related infections. PMID- 11379715 TI - Infection control dogma: top 10 suspects. AB - As infection control evolved into an art and science through the years, many infection control practices have become infection control dogmas (principles, beliefs, ideas, or opinions). In this "Reality Check" session of the 4th Decennial International Conference on Nosocomial and Healthcare-Associated Infections, we assessed participants' perceptions of prevalent infection control dogmas. The majority of participants agreed with all dogmas having evidence of efficacy, except for the dogma on the frequency of changing mechanical-ventilator tubing. In contrast, the majority of participants disagreed with dogmas not having evidence of efficacy, except for the dogma on perineal care, umbilical cord care, and reminder signs for isolation precaution. As for controversial dogmas, many of the responses were almost evenly distributed between "agree" and "disagree." Infection control professionals were knowledgeable about evidence based infection control practices. However, many of the respondents still believe in some of the non-evidence-based dogmas. PMID- 11379717 TI - The mortality of psychoanalysts. AB - The mortality rate for male psychoanalysts was compared to that for the general white male population; for male physicians; and for male psychiatrists and neurologists. For psychoanalysts the rate was found to be significantly lower than for any of the other three groups. Several possible explanations for this low mortality rate are considered. Two major factors may be the careful screening of candidates for psychoanalytic training and their personal analysis. Possible methods of controlling for these factors are suggested. PMID- 11379716 TI - A review of single-use and reusable gowns and drapes in health care. AB - Gowns and drapes are used widely in healthcare facilities. Gowns have been used to minimize the risk of disease acquisition by healthcare providers, to reduce the risk of patient-to-patient transmission, and during invasive procedures to aid in maintaining a sterile field. Drapes have been used during invasive procedures to maintain the sterility of environmental surfaces, equipment, and patients. This article reviews the use of gowns and drapes in healthcare facilities, including the characteristics, costs, benefits, and barrier effectiveness of single-use and reusable products. Currently, gowns protect healthcare personnel performing invasive procedures from contact with bloodborne pathogens. Although gowns have been recommended to prevent patient-to-patient transmission in certain settings (eg, neonatal intensive care unit) and for certain patients (eg, those infected with vancomycin-resistant enterococci), scientific studies have produced mixed results of their efficacy. While appropriate use of drapes during invasive procedures is recommended widely as an aid in minimizing contamination of the operative field, the efficacy of this practice in reducing surgical-site infections has not been assessed by scientific studies. Based on an evaluation of the functional requirements, environmental impact, and economics of gowns and drapes, clear superiority of either reusable or single-use gowns and drapes cannot be demonstrated. The selection of particular gowns and drapes by individual healthcare facilities requires an assessment of the facility's requirements, available products, and costs and should be based on the desired characteristics of an ideal gown or drape as defined in this paper. PMID- 11379718 TI - The American Psychoanalytic Association: a decade of change. AB - The American Psychoanalytic Association in the last decade has undergone the most extensive organizational changes in its history. Efforts at inclusion, outreach, and democratization have halted a decline in candidates, active members, and cultural influence and led to a vibrant, growing organization, as new professional groups have gradually been included in the association's training programs. The goal of increasing training opportunities for minority candidates has only partially been achieved. A recent effort to offer affiliate status to psychotherapists and students has begun, though with modest results. Alliances with otherpsychoanalytic groups have led to creation of the Psychoanalytic Consortium and steady progress toward external accreditation of institutes and licensure of psychoanalysts. In the interest of unification there is a need to develop pathways of affiliation and membership for independent institutes and societies that desire to join the association. Democracy has in large measure already been enhanced at the national level. Deliberations are now transparent, and an invigorated Executive Committee willingly shares power with the Executive Council. Candidates now are deeply involved in the governance of Board and Council and their committees. But only when all members and candidates enjoy full voting rights and all members are able to serve as officers and Executive Councilors will the decades-long effort to democratize the association be concluded. PMID- 11379719 TI - Mental representation, severe psychopathology, and the therapeutic process. AB - Mental representation is a central construct in psychological development. A method for assessing the developmental level of representation of self and significant figures is described, and changes in the developmental level of these representations are reported in a sample of forty seriously disturbed, treatment resistant adolescents and young adults in intensive, psychoanalytically oriented inpatient treatment lasting more than a year. Increased differentiation relatedness of descriptions of self and significant figures (mother, father, and therapist) was significantly correlated with improved clinical functioning. Over the course of treatment, representations moved from descriptions of self and significant figures dominated by polarization and splitting to representations involving the emergence and consolidation of object constancy. Improved clinical functioning was correlated with more positive descriptions of self, mother, and therapist and, paradoxically, with more negative descriptions of father. Two prototypical case studies of these self- and significant-figure descriptions are presented, one for a borderline patient and one for a schizophrenic. Intense negative affect, predominantly anger, and a relative preservation of self reflexivity are typical of the self- and object representations of borderline individuals, but representations in schizophrenic individuals are characterized by affective muting and marked disturbance in reflexive self-awareness. The assessment of cognitive-affective schemas of self and significant others provides a method for investigating therapeutic change and for identifying important differences among various forms of psychopathology. PMID- 11379720 TI - An empirical study of seriously disturbed suicidal patients. AB - Aspects of unconscious processes in a group of seriously disturbed psychiatric patients are examined in an effort to predict near-lethal suicide attempts and explore psychoanalytic formulations of suicide. The Rorschach Inkblot Test, the most widely used projective measure in suicide research (Bongar 1991), was chosen for its potential to shed light on specific unconscious processes. Psychic states commonly associated with suicide were measured by psychoanalytic Rorschach analog scales and then subjected to a progression of statistical analyses in order to predict future occurrence and lethality of suicide attempts. On the basis of a priori hypotheses, the authors developed a suicide index comprising four psychoanalytic Rorschach signs that predicted, with considerable accuracy, which patients would later make near-lethal suicide attempts. The best predictors were unconscious processes indicative of penetrating affective overstimulation, disturbance in the capacity to maintain adequate ego boundaries, and depressive affective states characterized by a morbid preoccupation with death and inner decay. These findings provide empirical support for several well-known formulations of the unconscious motivations for suicide. PMID- 11379721 TI - The appeal cycle in three cultures: an exploratory comparison of child development. AB - Mother-child behaviors were studied in three cultural groups: Caucasian American (CA), overseas Japanese (JPN), and Chinese Vietnamese (CVN) immigrants. The children were sixteen to twenty-five months old, and the appeal cycle, with its descriptive analytic method, was the research paradigm. Group differences were found in appeal cycle occurrence and phase frequencies. CA mothers promoted independence through encouragement of decision making and toddler-initiated play, and through nonintrusive interactions. Neither JPN nor CVN mothers encouraged independence. JPN mothers were subtly directive; CVN mothers overtly so. Generally, CA toddlers played independently, evincing a beginning capacity for self-regulation. JPN and CVN toddlers reacted to separation by staying close to their mothers, and fewer were observed to play independently than in the CA group. CVN toddlers played independently more frequently than JPN toddlers, but less frequently than CA toddlers. Quantitative measures of dyadic behaviors generally support the descriptive findings, though methodological constraints did not permit comparison of phase sequencing. The findings reflect cultural differences in child development thought to influence psychic structure formation. PMID- 11379722 TI - The Penn Psychoanalytic Treatment Collection: a set of complete and recorded psychoanalyses as a research resource. AB - From a set of seventeen complete and tape-recorded psychoanalyses, a sample of findings is presented: (a) the level of agreement of two clinical judges on the psychological health of these patients is adequate for the late sessions, but not for the early sessions; (b) the amount of change during psychoanalysis appears to be similar to that in the Menninger Foundation Psychotherapy Research Project; (c) psychiatric severity measures from the early sessions can yield a significant level of prediction of the later benefits from psychoanalysis. Finally, further research uses of this collection of psychoanalyses are suggested. PMID- 11379723 TI - The Columbia Supervision Project: data from the dyad. AB - Anonymous questionnaires were sent to all candidates and supervisors at the Columbia University Center for Psychoanalytic Training and Research (hereafter "Columbia"). Questions focused on the four domains most emphasized in the literature on supervision: logistical issues; the "teach or treat" question; the evaluatory function of the supervisor; and the affective experience of supervision. By coding the questionnaires, anonymity of respondents was maintained while allowing for a matched pair of analyses of supervisors and supervisees. Return rate was over 85 percent. In general, rates of satisfaction with supervision were high, and candidates and supervisors agreed on such issues as the "teach or treat" question, as well as the technical and theoretical frame of reference of the supervisor. However, there were striking disagreements between candidates and supervisors as to the role of the supervisor, what candidates find useful in supervision, the evaluatory function, and the relation between supervision and progression to graduation. Although 50 percent of candidates reported anxiety about receiving credit for cases, this was not routinely discussed in supervision, and the supervisory relationship itself was not discussed in over 50 percent of dyads. Despite high overall satisfaction ratings, 25 percent of candidates said they wished they had a different supervisor for the case, and 75 percent believed that a candidate who asked to switch supervisors would be labeled problematic. In contrast, over 75 percent of supervisors reported that switching supervisors carries no stigma. In a follow-up study conducted one year later, many candidates reported that they feared reprisals for switching, and some reported that their training analysts advised against "rocking the boat." Candidates felt that participating in the study emboldened them to think more openly about supervision and in some cases to make changes. PMID- 11379724 TI - Shame/contempt interchanges: a frequent component of the analyst-patient interaction. Panel report. PMID- 11379725 TI - Psychoanalysis and schizophrenia: a cautionary tale. AB - The history of psychoanalysis and schizophrenia is used as an example of psychoanalytic theories of etiology that have not stood the test of time. Those theories pointed to three main factors: very serious inadequacies in the caretaking person; the presence of these inadequacies so early, during the preverbal period, that they led to the impairment of early object relations, the development of psychic structure, and basic ego functions; and the absence of underlying biological abnormalities. Today, many analysts are still reluctant to acknowledge biological etiological factors for other psychiatric conditions. For illnesses such as borderline conditions and various severe character disorders for which biological factors are still much in doubt, analysts are today proposing etiological formulations similar to those once advanced for schizophrenia. These formulations may indeed prove correct for these disturbances, but analysts are urged to heed the cautionary tale of psychoanalysis and schizophrenia. PMID- 11379726 TI - Diagnosing The English Patient: schizoid fantasies of being skinless and of being buried alive. AB - The psychological world of The English Patient is explored to deepen the understanding of schizoid states. The protagonist, Almasy, is a remote desert explorer whose triangular sadomasochistic affair with the married Katharine destroys them all. His damaged skin is understood as a symbolic representation of his psychological condition. For the schizoid, love consumes and leads to obliteration of the self, represented by the loss of identifying features, and to traumatic permeability (i.e., the loss of boundaries between self and other, and between the ego and repressed desires). Other schizoid themes are the animation of the inanimate, as in the depiction of the desert as a woman; hidden or buried identities; the digital and destructive experience of emotion represented by the conundrum of the bomb defuser; the sense that everything good is imaginary and might suddenly explode; and the moral unevenness of the characters. Almasy collaborates with the Nazis so he can retrieve Katharine's three-year-old corpse, with which he has necrophilic contact in a cave. Fantasies of the lost object buried within the self, of being buried alive, and of being skinned alive are related to the schizoid condition. Hyperpermeability is proposed as a core schizoid state, underlying schizoid withdrawal. PMID- 11379727 TI - JAPA on the cutting edge. PMID- 11379728 TI - Toward a "psychodynamic science": the state of current research. PMID- 11379729 TI - Drug dreams: an introduction. PMID- 11379730 TI - Drug dreams: a neuropsychoanalytic hypothesis. AB - Recent studies have shown that the ventral tegmental pathway stimulates both dreaming and drug craving. To investigate a possible clinical link between these two psychic phenomena, psychotherapy notes from the first six months of an addicted patient's treatment were reviewed, together with verbatim notes from the four years of psychoanalysis that followed. Of 240 dreams reported by the patient,58 had manifest content involving the seeking or using of drugs. There was no particular temporal or emotional thematic pattern to these "drug dreams,"which persisted through four and a half years of sobriety. Drug dreams are observable phenomena that reflect both the innate structure of the brain and neural changes produced by exposure to addictive drugs. In some addicted persons, exposure to drugs produces a fixed change in neurological functioning with which they must contend for years, possibly the rest of their lives. Drug craving meets Freud's defining characteristics for a drive: it is a constant pressure, originating from within the organism, to do work, and it constantly demands satisfaction. Because ego and libidinal drives share a common neural pathway, they should not be separated conceptually. Solms's finding (in press) that the activating systems for dreaming and for craving are identical, a finding based on observations of tumor- or stroke-provoked brain lesions, is confirmed by observation of the dreams of a patient whose brain changes were created by drug exposure. This study provides further evidence that the origin of the dream is a wish. PMID- 11379731 TI - Introduction to Jeffery: why psychoanalysts have low mortality rates. PMID- 11379732 TI - Reverse obliquity fractures of the intertrochanteric region of the femur. AB - BACKGROUND: The reverse obliquity fracture of the proximal part of the femur is a distinct fracture pattern that is mechanically different from most intertrochanteric fractures. The purpose of this retrospective study was to determine the prevalence of these fractures and the results and complications of different types of internal fixation used in their treatment. METHODS: Between 1988 and 1998, 2472 consecutive patients with a hip fracture were treated at our Level-One Trauma Center; 1035 of the fractures were classified as intertrochanteric or subtrochanteric. Clinical and radiographic records were retrospectively reviewed, and fifty-five fractures with a reverse obliquity pattern were identified. Forty-nine patients were followed until the fracture united or a revision operation was performed. The duration of clinical follow-up averaged eighteen months (range, three to sixty-seven months), and the duration of radiographic follow-up averaged fifteen months (range, three to sixty months). Fractures were classified with the Orthopaedic Trauma Association scheme. Results were analyzed according to the fracture pattern, type of implant, quality of the reduction, position of the implant, and use of bone graft at the index operation. Function was assessed on the basis of pain, living situation, need for walking aids, need for analgesics, and walking capacity. RESULTS: Thirty-two (68%) of forty-seven hips treated with internal fixation healed without an additional operation. Fifteen (32%) of the forty-seven failed to heal or had a failure of fixation. The failure rate was nine of sixteen for the sliding hip screws, two of fifteen for the blade-plates, three of ten for the dynamic condylar screws, one of three for the cephalomedullary nails, and zero of three for the intramedullary hip screws. Use of the fixed-angle devices (the blade-plate and the dynamic condylar screw) resulted in fewer failures than did use of the sliding hip screw (p = 0.023). Eleven (46%) of twenty-four nonanatomically reduced fractures and four (17%) of twenty-three anatomically reduced fractures had a failure of treatment (p = 0.060). Eleven (26%) of forty-two fractures with an ideally placed implant and four (80%) of five fractures with a non-ideally placed implant had a failure of treatment (p = 0.023). Of the fifteen fractures that failed to heal or had a failure of fixation, five were treated with revision to a calcar replacement prosthesis, seven were treated with revision open reduction and internal fixation with bone-grafting, and one was treated with bone-grafting without revision of the fixation. Two patients refused additional surgery because they had limited functional demands. The two-year mortality rate was 33%. Functional results were poor, with many patients requiring walking aids and losing the capacity for independent walking and self-care. CONCLUSIONS: In this series, reverse obliquity fractures accounted for 2% of all hip fractures and 5% of all intertrochanteric and subtrochanteric fractures. Ninety-five-degree fixed angle internal fixation devices performed significantly better than did sliding hip screws. Results were also worse for fractures with poor reduction and those with a poorly placed implant. PMID- 11379733 TI - Fluorine-18 fluorodeoxyglucose-position emission tomography: a highly accurate imaging modality for the diagnosis of chronic musculoskeletal infections. AB - BACKGROUND: The noninvasive diagnosis of chronic musculoskeletal infections remains a challenge. Recent studies have indicated that fluorine-18 fluorodeoxyglucose-positron emission tomography is a highly accurate imaging technique and is significantly more accurate than the combination of a bone scan and a white blood-cell scan for the diagnosis of chronic infection in the central skeleton (p < 0.05). However, patients who had had surgery within the previous two years were excluded from study. It was our aim to evaluate the technique in an unselected, clinically representative population. METHODS: Sixty patients with a suspected chronic musculoskeletal infection involving the central skeleton (thirty-three patients) or the peripheral skeleton (twenty-seven patients) were studied with fluorine-18 fluorodeoxyglucose-positron emission tomography. Thirty five patients had had surgery within the previous two years. The fluorine-18 fluorodeoxyglucose-positron emission tomography studies were read in a blinded, independent manner by two experienced readers. The final diagnosis was based on histopathological studies or microbiological culture (eighteen patients) or on clinical findings after at least six months of follow-up (forty-two patients). RESULTS: On the final composite assessment, twenty-five patients had infection and thirty-five did not. All twenty-five infections were correctly identified by both readers. There were four false-positive findings; in two of these cases, surgery had been performed less than six months prior to the study. The sensitivity, specificity, and accuracy were 100%, 88%, and 93% for the whole group; 100%, 90%, and 94% for the subgroup of patients with a suspected infection of the central skeleton; and 100%, 86%, and 93% for the subgroup of patients with a suspected infection of the peripheral skeleton. Interobserver agreement was excellent (kappa = 0.97). CONCLUSIONS: Fluorine-18 fluorodeoxyglucose-positron emission tomography is highly accurate as a single technique for the evaluation of chronic musculoskeletal infections. It is especially valuable in the evaluation of the central skeleton, where white blood-cell scans are less useful. Because of its simplicity and high degree of accuracy, it has the potential to become a standard technique for the diagnosis of chronic musculoskeletal infections. Further studies are needed to assess its ability to identify infections at the sites of total joint replacements and to distinguish infection from aseptic loosening of these prostheses. PMID- 11379734 TI - Position of immobilization after dislocation of the glenohumeral joint. A study with use of magnetic resonance imaging. AB - BACKGROUND: Glenohumeral dislocations often recur, probably because a Bankart lesion does not heal sufficiently during the period of immobilization. Using magnetic resonance imaging, we assessed the position of the Bankart lesion, with the arm in internal and external rotation, in shoulders that had had a dislocation. METHODS: Coaptation of a Bankart lesion was examined with use of magnetic resonance imaging, with the arm held at the side of the trunk and positioned first in internal rotation (mean, 29 degrees) and then in external rotation (mean, 35 degrees), in nineteen shoulders. Six shoulders (six patients) had had an initial anterior dislocation, and thirteen shoulders (twelve patients) had had recurrent anterior dislocation. Fast-spin-echo T2-weighted axial images were made when the dislocation had occurred less than two weeks earlier, and spin echo T1-weighted axial images after intra-articular injection of gadolinium diethylenetriamine pentaacetic acid were made when the dislocation had occurred more than two weeks earlier. Separation and displacement of the anteroinferior portion of the labrum from the glenoid rim were measured on the axial images, and coaptation of the anterior part of the capsule to the glenoid neck was assessed by measurement of the detached area, opening angle, and detached length. RESULTS: Separation and displacement of the labrum were both significantly less (p = 0.0047 and p = 0.0017, respectively) when the arm was in external rotation than when it was in internal rotation. The detached area and the opening angle of the anteroinferior portion of the capsule were both significantly smaller (p = 0.0003 and p < 0.0001, respectively), and the detached length was significantly shorter (p < 0.0001) with the arm in external rotation. CONCLUSION: Immobilization of the arm in external rotation better approximates the Bankart lesion to the glenoid neck than does the conventional position of internal rotation. PMID- 11379735 TI - Impact of smoking on the outcome of anterior cervical arthrodesis with interbody or strut-grafting. AB - BACKGROUND: An increased rate of pseudarthrosis has been documented following posterolateral lumbar spine grafting in patients who smoke. This same relationship has been assumed for anterior cervical interbody grafting, but to our knowledge it has never been proven. This study compared the long-term radiographic and clinical results of smokers and nonsmokers who had undergone arthrodesis with autogenous bone graft following multi-level anterior cervical decompression for the treatment of cervical radiculopathy or myelopathy, or both. METHODS: One hundred and ninety patients were followed clinically and radiographically for at least two years (range, two to fifteen years). Fifty-nine of the patients had corpectomy with strut-grafting, and 131 patients had multiple discectomies and interbody grafting. Fifty-five of the 190 patients had a history of active cigarette-smoking; fifteen of the fifty-five had corpectomy with strut grafting, and forty had multilevel discectomies and interbody grafting. Internal fixation was not used in any patient. The reconstruction techniques and postoperative bracing regimen were similar between smokers and nonsmokers. Osseous union was judged on dynamic lateral radiographs made at least two years following surgery, and clinical outcomes were judged on the basis of pain level, medication usage, and daily activity level. RESULTS: Of the forty smokers who had undergone multilevel interbody grafting, twenty had a solid fusion at all levels, whereas sixty-nine of the ninety-one nonsmokers had solid fusion at all levels (p < 0.02; chi-square test). This difference was especially pronounced among patients who had had a two-level interbody grafting procedure (p < 0.002; chi square test). With the numbers available, there was no difference in the rate of fusion between smokers (fourteen of fifteen) and nonsmokers (forty-one of forty four) who had undergone corpectomy and strut-grafting, as 93% of both groups had a solid union. In addition, clinical outcomes were significantly worse among smokers when compared with nonsmokers (p < 0.03; rank-sum analysis). CONCLUSIONS: Smoking had a significant negative impact on healing and clinical recovery after multilevel anterior cervical decompression and fusion with autogenous interbody graft for radiculopathy or myelopathy. Since smoking had no apparent effect upon the healing of autogenous iliac-crest or fibular strut grafts, subtotal corpectomy and autogenous strut-grafting should be considered when a multilevel anterior cervical decompression and fusion is performed in patients who are unable or unwilling to stop smoking prior to surgical treatment. PMID- 11379736 TI - Radiation exposure with use of the inverted-c-arm technique in upper-extremity surgery. AB - BACKGROUND: Intraoperative fluoroscopy is commonly used in surgical procedures on upper extremities. We compared radiation exposure from two possible positions of the mobile digital fluoroscopy unit (c-arm): (1) the standard technique, with the x-ray tube down (near the floor) and the image intensifier at the top of the c arm, and (2) the inverted position, in which the image intensifier is used as a table and the x-ray tube is up. METHODS: A commercially available c-arm was used to irradiate a phantom hand in one of three configurations. In the first, the phantom hand was placed on an armboard equidistant from the x-ray tube and the image intensifier with the beam directed upward. In the second, the c-arm was inverted with the beam directed downward and the image intensifier used as a table. The third configuration was identical to the second except that a magnified image was used. Radiation exposure was measured at four locations corresponding to the approximate position of the surgeon's head, chest, and groin and the patient's hand. RESULTS: The amount of radiation exposure to both the surgeon and the patient was significantly less when the c-arm was used in the inverted position (p < 0.0001). The dose rate to the patient's hand was reduced by 59%. The radiation exposure to the surgeon's head, body, and groin with the inverted-c-arm technique was 67%, 45%, and 15% of the measured doses with the x ray-tube-down configuration. When we used the magnification mode of the image intensifier, with its correspondingly smaller field size, the doses were further reduced to 46%, 32%, and 11% of the standard-configuration values. CONCLUSIONS: Use of the inverted-c-arm technique with the image intensifier as an operating table can significantly reduce radiation exposure to the surgeon and the patient during surgical procedures on upper extremities. PMID- 11379737 TI - Clinical determinants of the outcome of manual stretching in the treatment of congenital muscular torticollis in infants. A prospective study of eight hundred and twenty-one cases. AB - BACKGROUND: The natural history of congenital muscular torticollis and the outcome of different treatment modalities have been poorly investigated, and the results of treatment have varied considerably. METHODS: The main objective of this prospective study was to evaluate the outcomes of 821 consecutive patients with congenital muscular torticollis who were first seen when they were less than one year old, were treated with a standardized program of manual stretching, and were followed for a mean of 4.5 years. Before treatment, the patients were classified into one of three clinical groups: (1) palpable sternomastoid tumor, (2) muscular torticollis (thickening and tightness of the sternocleidomastoid muscle), and (3) postural torticollis (torticollis but no tightness or tumor). RESULTS: Of the 821 patients, 452 (55%) had a sternomastoid tumor; 276 (34%), muscular torticollis; and ninety-three (11%), postural torticollis. Multivariate analysis of the outcomes showed that (1) the duration of treatment was significantly associated with the clinical group (p < 0.0001), a passive rotation deficit of the neck (p < 0.0001), involvement of the right side (p < 0.0001), difficulties with the birth (p < 0.009), and age at presentation (p < 0.0001); (2) the overall final assessment score was associated with the rotation deficit (p = 0.02), age at presentation (p = 0.014), and duration of treatment (p < 0.0001); and (3) subsequent surgical treatment was required by 8% (thirty-four) of the 452 patients in the sternomastoid tumor group compared with 3% (eight) of the 276 patients in the muscular torticollis group and 0% (none) of the ninety three patients in the postural torticollis group. CONCLUSIONS: This large prospective study demonstrated that controlled manual stretching is safe and effective in the treatment of congenital muscular torticollis when a patient is seen before the age of one year. The most important factors that predict the outcome of manual stretching are the clinical group, the initial deficit in rotation of the neck, and the age of the patient at presentation. Surgical treatment is indicated when a patient has undergone at least six months of controlled manual stretching and has residual head tilt, deficits of passive rotation and lateral bending of the neck of >15 degrees, a tight muscular band or tumor, and a poor outcome according to our special assessment chart. PMID- 11379739 TI - Arthroscopically assisted treatment of avulsion fractures of the posterior cruciate ligament from the tibia. AB - BACKGROUND: The attachment of the posterior cruciate ligament to the posterior intercondylar fossa of the tibia is in a location that is difficult to access for arthroscopic surgical procedures. This report presents a variety of arthroscopically assisted reduction and fixation methods for managing avulsion fractures of the posterior cruciate ligament from the tibia. METHODS: Thirteen patients (fourteen knees) who had an avulsion fracture of the posterior cruciate ligament were treated with an arthroscopic procedure. Eleven patients underwent the operation in the acute phase (four to ten days after the injury), and two patients had delayed surgery (at nineteen and twenty months after the injury) because of nonunion. The choice of fixation method was based on the size of the avulsed fragment. Six knees that had a small bone fragment (<10 mm) with comminution were fixed with use of multiple sutures. Two knees that had a small bone fragment without comminution were fixed with 23-gauge wires. Two knees that had a medium-sized fragment (10 to 20 mm) were fixed with Kirschner wires. Four knees that had a large single fragment of bone (>20 mm) that involved the condyles were fixed with one or two cannulated screws. RESULTS: All patients had osseous union as determined on radiographs. Three injured knees in two patients showed limitation of motion after the operation. These patients had been immobilized for two or three months after the surgery because of concomitant fractures. The eleven patients who had undergone the operation in the acute phase, including two in whom postoperative arthrofibrosis had developed, showed no or trace posterior instability following the procedure. However, the two patients in whom the surgery had been delayed had residual grade-I posterior instability. The postoperative side-to-side differences, when measured with use of the KT-2000 arthrometer and posterior stress radiographs, showed better results in the patients in whom the surgery had been performed in the acute phase than in the patients in whom the operation had been delayed. CONCLUSION: Arthroscopic procedures can be used to treat tibial avulsion fractures of the posterior cruciate ligament. PMID- 11379738 TI - Histopathologic changes in growth-plate cartilage following ischemic necrosis of the capital femoral epiphysis. An experimental investigation in immature pigs. AB - BACKGROUND: The developing capital femoral epiphysis consists of a secondary center of ossification surrounded by epiphyseal cartilage. Between the epiphyseal cartilage and the secondary center of ossification is a growth plate, which contributes to the circumferential increase in size of the secondary center of ossification during development. The main objective of this study was to describe the histopathologic changes that occur in the growth plate surrounding the secondary center of ossification during the early and reparative phases following the induction of ischemic necrosis of the capital femoral epiphysis in immature pigs. METHODS: Ischemic necrosis of the capital femoral epiphysis was induced in eighteen piglets by placing a nonabsorbable suture ligature around the femoral neck following a capsulotomy and transection of the ligamentum teres. The animals were killed three days to eight weeks following the induction of ischemia, and visual, radiographic, and histologic assessments were performed. RESULTS: Two to four weeks after the induction of ischemic necrosis, the growth plate surrounding the secondary center of ossification became necrotic. The observed histopathologic changes included chondrocyte death, loss of safranin-O staining of the matrix of the necrotic growth-plate cartilage, an absence of vascular invasion of terminal hypertrophic chondrocytes, and a decrease in the amount of primary spongiosa, indicating cessation of endochondral ossification. In the reparative phase, at four to eight weeks postoperatively, chondrocyte clusters and intense safranin-O staining were observed in the epiphyseal cartilage around the necrotic growth-plate cartilage. In the peripheral region of the femoral head, necrotic growth-plate cartilage surrounding the secondary center of ossification was resorbed by a fibrovascular tissue from the marrow space. By six weeks, new accessory centers of ossification with restored endochondral ossification were observed in the peripheral epiphyseal cartilage. New ossification centers contributed to the fragmented radiographic appearance of the secondary center of ossification. The physis appeared essentially normal in most animals, although five of the eighteen piglets showed mild or moderate histopathologic changes. CONCLUSIONS: In this model, ischemic necrosis of the capital femoral epiphysis resulted in necrosis of the growth plate surrounding the secondary center of ossification. Small new ectopic centers of ossification appeared in the epiphyseal cartilage, explaining in part the fragmented radiographic appearance of the secondary center of ossification. PMID- 11379740 TI - Influence of metacarpophalangeal joint position on basal joint-loading in the thumb. AB - BACKGROUND: Conventional wisdom holds that hyperextension of the metacarpophalangeal joint of the thumb is secondary to degenerative subluxation of the trapeziometacarpal joint as occurs in osteoarthritis. We propose that a hypermobile metacarpophalangeal joint may have a causative role in the development of primary osteoarthritis at the base of the thumb by concentrating forces on the palmar aspect of the trapeziometacarpal joint. METHODS: Twenty fresh-frozen cadaveric forearm specimens were obtained post mortem from donors with no history of connective-tissue disease. Each specimen was categorized by its passive range of metacarpophalangeal joint motion. Testing was conducted with Fuji ultra-low-pressure-sensitive film while the hand was in the lateral-pinch mode with the metacarpophalangeal joint in each of the following positions: unrestrained, pinned in neutral, pinned in 30 degrees of flexion, and pinned in maximal hyperextension. Quantitative analysis of the trapezial contact surface at each of the metacarpophalangeal joint positions was performed, and the center of pressure was determined. Each specimen was then classified according to the extent of arthritic disease (nonarthritic, moderately arthritic, or affected by end-stage arthritis). RESULTS: In specimens affected by end-stage osteoarthritis, the center of pressure on the trapeziometacarpal joint moved dorsally by 56.8% of the length of the trapezial surface with metacarpophalangeal joint flexions of 30 degrees (p < 0.01), whereas the corresponding values were 28.2% and 40.9% in the hyperextended and neutral metacarpophalangeal joint positions, respectively. In specimens with moderate osteoarthritis, 30 degrees of metacarpophalangeal joint flexion also produced the most dorsal trapeziometacarpal center of pressure (44.8%); however, this center of pressure was not significantly different from the centers of pressure at the other metacarpophalangeal joint positions. In nonarthritic specimens, the center of pressure was again significantly more dorsal with metacarpophalangeal joint flexion of 30 degrees than it was at the other positions (p < 0.01). CONCLUSION: Metacarpophalangeal joint flexion effectively unloaded the most palmar surfaces of the trapeziometacarpal joint regardless of the presence or severity of arthritic disease in this joint. PMID- 11379742 TI - A brief note. Ask yourself, why? A cost effective look at routine pathologic examination of specimens using the trapezium. PMID- 11379741 TI - Improvement of the bone-pin interface strength in osteoporotic bone with use of hydroxyapatite-coated tapered external-fixation pins. A prospective, randomized clinical study of wrist fractures. AB - BACKGROUND: Achieving adequate fixation strength in osteoporotic bone is a challenge. In this study, we examined the use of hydroxyapatite-coated tapered external-fixation pins for the fixation of wrist fractures in patients with osteoporosis. METHODS: Twenty female patients with osteoporosis and a fracture of the wrist were divided into two paired groups and randomized to receive either standard tapered pins or hydroxyapatite-coated tapered pins. Two pins were inserted in the distal part of the radius, two pins were inserted in the second metacarpal, and an external fixation device was mounted. All fixation devices were removed six weeks after surgery. RESULTS: The mean pin-insertion torque (and standard deviation) was 461 +/- 254 Nmm in the group managed with standard pins and 332 +/- 176 Nmm in the group managed with hydroxyapatite-coated pins (p = 0.01). The mean pin-extraction torque was 191 +/- 155 Nmm in the group managed with standard pins and 600 +/- 214 Nmm in the group managed with hydroxyapatite coated pins (p < 0.0001, power 95%). The mean extraction torque was lower than the corresponding insertion torque at each pin position in the group managed with standard pins (p < 0.05), whereas the mean extraction torque was higher than the corresponding insertion torque at each pin position in the group managed with hydroxyapatite-coated pins (p = 0.001). Two patients managed with standard pins and no patient managed with hydroxyapatite-coated pins had a pin-track infection. Pain during pin removal did not differ between the two groups. CONCLUSIONS: The present study showed that hydroxyapatite-coated tapered external-fixation pins provided improved fixation in the treatment of wrist fractures in patients with osteoporosis. PMID- 11379743 TI - The effect of placing a tensioned graft across open growth plates. A gross and histologic analysis. AB - BACKGROUND: Midsubstance tears of the anterior cruciate ligament in skeletally immature patients are increasingly common and are a challenging problem. The results of nonoperative treatment are no better in children than they are in adults. Physeal-sparing reconstructive procedures have yielded poor results. Reconstructive procedures that are utilized in adults violate the physis, potentially resulting in growth abnormalities. The objective of this study was to provide a model for reconstruction of the anterior cruciate ligament in skeletally immature patients by evaluating the effects of a tensioned connective tissue graft placed across the canine physis. METHODS: Twelve ten-week-old beagles underwent reconstruction of the anterior cruciate ligament consisting of placement of fascia lata autograft through drill-holes across the femoral and tibial physes, tensioning of the graft to 80 N, and fixing it with screws and washers. The contralateral limb served as a control. One dog was eliminated from the study secondary to a postoperative infection. Four months postoperatively, the dogs were killed and were inspected grossly, radiographically, and histologically for any evidence of growth disturbance. RESULTS: Significant valgus deformity of the distal part of the femur (p < 0.001) and significant varus deformity of the proximal part of the tibia (p = 0.03) developed in the treated limbs. Neither radiographic nor histologic examination demonstrated any evidence of physeal bar formation. CONCLUSIONS: Significant growth disturbances occur with excessively tensioned transphyseal reconstruction of the anterior cruciate ligament in the canine model. These growth disturbances occur without radiographic or histologic evidence of physeal bar formation. PMID- 11379744 TI - Operative treatment of supracondylar fractures of the humerus in children. The consequences of pin placement. AB - BACKGROUND: The commonly accepted treatment of displaced supracondylar fractures of the humerus in children is fracture reduction and percutaneous pin fixation; however, there is controversy about the optimal placement of the pins. A crossed pin configuration is believed to be mechanically more stable than lateral pins alone; however, the ulnar nerve can be injured with the use of a medial pin. It has not been proved that the added stability of a medial pin is clinically necessary since, in young children, pin fixation is always augmented with immobilization in a splint or cast. METHODS: We retrospectively reviewed the results of reduction and Kirschner wire fixation of 345 extension-type supracondylar fractures in children. Maintenance of fracture reduction and evidence of ulnar nerve injury were evaluated in relation to pin configuration and fracture pattern. Of 141 children who had a Gartland type-2 fracture (a partially intact posterior cortex), seventy-four were treated with lateral pins only and sixty-seven were treated with crossed pins. Of 204 children who had a Gartland type-3 (unstable) fracture, fifty-one were treated with lateral pins only and 153 were treated with crossed pins. RESULTS: There was no difference with regard to maintenance of fracture reduction, as seen on anteroposterior and lateral radiographs, between the crossed pins and the lateral pins. The configuration of the pins did not affect the maintenance of reduction of either the Gartland type-2 fractures or the Gartland type-3 fractures. Ulnar nerve injury was not seen in the 125 patients in whom only lateral pins were used. The use of a medial pin was associated with ulnar nerve injury in 4% (six) of 149 patients in whom the pin was applied without hyperflexion of the elbow and in 15% (eleven) of seventy-one in whom the medial pin was applied with the elbow hyperflexed. Two years after the pinning, one of the seventeen children with ulnar nerve injury had persistent motor weakness and a sensory deficit. CONCLUSIONS: Fixation with only lateral pins is safe and effective for both Gartland type-2 and Gartland type-3 (unstable) supracondylar fractures of the humerus in children. The use of only lateral pins prevents iatrogenic injury to the ulnar nerve. On the basis of our findings, we do not recommend the routine use of crossed pins in the treatment of supracondylar fractures of the humerus in children. If a medial pin is used, the elbow should not be hyperflexed during its insertion. PMID- 11379745 TI - Surgical repair of distal biceps tendon rupture complicated by median nerve entrapment. A case report. PMID- 11379747 TI - Gaucher disease. New approaches to an ancient disease. PMID- 11379746 TI - Primary hyperparathyroidism: an unusual cause of cervical myelopathy. A case report. PMID- 11379748 TI - Medical scientific publishing in the twenty-first century. Synopsis of symposium presented at the Annual Meeting of the American Orthopaedic Association. PMID- 11379749 TI - The risk of cancer following total hip or knee arthroplasty. PMID- 11379750 TI - Routine pathological examination of specimens obtained during total joint replacement. PMID- 11379751 TI - Workers' compensation: the patient, the physician, and the system. PMID- 11379752 TI - Prosthetic design and outcome in total hip arthroplasty. PMID- 11379753 TI - Total knee arthroplasty after high tibial osteotomy. PMID- 11379754 TI - Detecting valgus instability in the elbow. PMID- 11379755 TI - Intermittent pneumatic compression promoted healing in foot infections. PMID- 11379756 TI - What's new in foot and ankle surgery. PMID- 11379758 TI - Proteolytic activation and inactivation of the serine protease activity of plasma hyaluronan binding protein. AB - We prepared anti-plasma hyaluronan binding protein (PHBP) mouse monoclonal antibodies and studied the fragmentation profile of PHBP with them. PHBP is present in human plasma as a single polypeptide chain (70 kDa). During the purification, PHBP partially fragmentated into the 50-kDa N-terminal fragment and the 27-kDa C-terminal fragment. After the incubation of the purified PHBP, the 70 kDa precursor form was completely cleaved to the 50- and 27-kDa fragments, followed by the 50-kDa to the 26-kDa, and the 27-kDa to the 17-kDa plus the 8-kDa fragments, respectively. Because the purified PHBP contained no other detectable proteins and PHBP has a typical serine protease domain, we concluded that the fragmentation of PHBP was caused by own serine protease activity. PHBP cleaved the C-terminal side of Arg in the peptide effectively and that of Lys weakly. The results of the pre-incubation experiments of PHBP suggested that the single-chain form of PHBP is a precursor, the two-subunit structure is an active form and the three- or four-chain structure is an inactive form of a serine protease. PMID- 11379757 TI - Characterization of bovine angiogenin-1 and lactogenin-like protein as glycyrrhizin-binding proteins and their in vitro phosphorylation by C-kinase. AB - Angiogenin-1 (p15, an angiogenesis inducer with RNase activity) and lactogenin like protein (p17) isolated from partially purified bovine lactoferrin (bLF) preparations were characterized as glycyrrhizin (GL)-binding proteins (gbPs). As expected, bLF-affinity column chromatography confirmed these two gbPs to be bLF binding proteins. These two purified gbPs exhibited RNase activities when incubated with poly(C) as a substrate. Both GL and glycyrrhetinic acid (GA) at 100 microM significantly inhibited RNase activities of these two gbPs, both of which functioned as phosphate acceptors of C-kinase in vitro. Phosphorylation of p15 and p17 by C-kinase was inhibited by GA in a dose-dependent manner with the 50% inhibition dose (ID50) of approx. 10 microM, whereas GL required a relatively high dose (300 microM) to inhibit significantly it. A GA derivative (oGA, ID50=approx. 0.3 microM) was found to be a potent inhibitor of the C-kinase mediated phosphorylation of these two gbPs in vitro. In addition, a possible physiological significance of C-kinase on the physiological interaction between bLF and two bLF-binding proteins (p15 and p17) is noted. PMID- 11379759 TI - Cyclic-AMP inhibits nitric oxide-induced apoptosis in human osteoblast: the regulation of caspase-3, -6, -9 and the release of cytochrome c in nitric oxide induced apoptosis by cAMP. AB - Nitric oxide (NO) induces apoptotic cell death and cAMP has a significantly protective effect on NO-induced cytotoxicity in human osteoblasts, MG-63 cells. Treatment with S-nitroso-N-acetylpenicillamine (SNAP) (0.6 mM) resulted in genomic DNA fragmentation, characteristic of apoptosis. However, concomitant incubation of the cells with either DBcAMP or forskolin markedly inhibited SNAP induced apoptosis in a dose-dependent manner. Furthermore, pretreatment of MG-63 cells with H-89 or KT5720, which is known to inhibit cAMP-dependent protein kinase (PKA), abolished the protective effect of DBcAMP and forskolin on SNAP induced apoptosis. In this study, we explored the involvement of caspases in the regulatory mechanism of SNAP-induced apoptosis by cAMP. Our data show that DBcAMP or forskolin blocked SNAP-induced caspase-3-like cysteine protease activation and that H-89, a PKA inhibitor, reversed the cAMP-induced regulatory effect of caspase-3 like protease. Consistent with the results, cAMP inhibited the proteolytic cleavage of caspase-3, -6, -9 and cytochrome c release to cytoplasm. The inhibition of caspase-3 activation did not block SNAP-induced cytochrome c release to cytoplasm, suggesting that caspase-3 activation may occur downstream of cytochrome c release. In summary, these findings show that the exposure of MG 63 cells to cAMP analogs renders them more resistant to NO-induced damage and suggests the presence of regulatory mechanisms of the cell death pathway by cAMP in which caspase-3, -6, and -9 and cytochrome c release serves to mediate NO induced apoptosis. PMID- 11379760 TI - Nitric oxide stimulates elastin expression in chick aortic smooth muscle cells. AB - Nitric oxide (NO), an endothelium-dependent relaxing factor, regulates relaxation, proliferation, and migration of smooth muscle cells (SMCs) and most likely attenuates developing vascular disease such as atherosclerosis. We investigated whether or not NO is associated with regulation of aortic elasticity. S-Nitrosoglutathione (GSNO), a NO donor, stimulated tropoelastin synthesis in cultured SMCs during both the quiescent and proliferating phases. The stimulation of tropoelastin synthesis was dose-dependent within 1-100 nM. Maximum stimulation was detected by treatment with 100 nM GSNO for 24 h. 8 Bromoguanosine 3',5'-cyclic monophosphate (8-Br-cGMP), an exogenous cyclic GMP analog, also upregulated tropoelastin synthesis. Tropoelastin and lysyl oxidase mRNA expression, as assessed by Northern blot analysis, was also stimulated by GSNO. Administration of KT5823, a cyclic GMP-dependent protein kinase inhibitor, inhibited the GSNO-induced tropoelastin synthesis. These results indicate that the stimulatory effects of GSNO are due to cyclic GMP dependent protein kinase (PKG) activation by NO. In conclusion, NO seems to enhance aortic elasticity via tropoelastin and lysyl oxidase upregulation. PMID- 11379761 TI - Effects of dermatan and dextran sulfates on arginine amidase activity released from isolated rabbit arteries. AB - We examined the levels of arginine amidase secreted from isolated rabbit arteries treated with dermatan and dextran sulfates, and the relation between the secretion of arginine amidase activity and the concentration of dermatan sulfate added to these arteries. The results showed that while dextran sulfate tended to accelerate the release of arginine amidase activity from the isolated rabbit ear artery, the induction was not significant. There was a significant increase in the level of arginine amidase released from the lower portion of isolated rabbit aorta (p<0.05), but no significant change in the upper portion of the aorta. In contrast, the addition of dermatan sulfate significantly increased the level of arginine amidase activity released from the isolated rabbit ear artery and the upper and lower portions of the aorta (p<0.05). Linear dose-response relationships were observed between the level of arginine amidase activity released from the isolated rabbit ear artery and aorta and the concentration of dermatan sulfate added. PMID- 11379762 TI - Anti-inflammatory and immunomodulating properties of a sterol fraction from Sideritis foetens Clem. AB - A sterol fraction composed of campesterol (7.6%), stigmasterol (28.4%) and beta sitosterol (61.1%) was obtained by activity-guided fractionation of the acetone extract of Sideritis foetens Clem. This sterol fraction showed anti-inflammatory activity in in vivo murine models of inflammation. It decreased carrageenan paw oedema in mice after oral administration of 30 and 60 mg/kg and inhibited mouse ear oedema induced by 12-O-tetradecanoylphorbol acetate (TPA) after topical application. Quantitation of the neutrophil specific marker myeloperoxidase (MPO) demonstrated that its topical anti-inflammatory activity was associated with reduction in neutrophil infiltration into inflamed tissues. Non-cytotoxic concentrations of the sterol fraction inhibited leukocyte granular enzyme release (beta-glucuronidase) and superoxide generation. However, it did not shown any significant inhibitory effect on histamine release from mast cells. In vitro modulatory activity towards the classical pathway of the complement system shown by this fraction would correlate with the anti-inflammatory profile shown in vivo. PMID- 11379763 TI - Mechanism of resistance to oxidative stress in doxorubicin resistant cells. AB - Doxorubicin (DOX) is an anthracycline drug widely used in chemotherapy for cancer patients, but it often gives rise to multidrug resistance in cancer cells. The purpose of this work was to study the effect of hydrogen peroxide in DOX sensitive mouse P388/S leukemia cells and in the DOX-resistant cell line. Hydrogen peroxide induced a significant increase in dose- and time-response cell death in cultured P388/S cells. The degree of cell death in P388/DOX cells induced by hydrogen peroxide was less than that in P388/S cells treated with hydrogen peroxide. Parent cells exposed to 3 mM of hydrogen peroxide showed a loss of mitochondrial membrane potential correlated with cell death. Hydrogen peroxide at a concentration greater than 0.3 mM increased the intracellular Ca2+ of P388/S cells dose-dependently; however, no change following addition of hydrogen peroxide (0.3-1 mM) was observed in the resistant cells. Hydrogen peroxide (0.1 and 1 mM) treatment also induced the production of intracellular ROS in P388/S cells, while no such increase was produced by this substance in P388/DOX cells. Resistant cells also showed a significant level of glutathione (GSH) compared with the parent cells. In addition, N-acetyl-L-cysteine and reduced GSH antioxidants abolished death of P388/S cells caused by hydrogen peroxide. Therefore, it is believed that the reduced effect of oxidative stress towards the resistant cells may be related to an increase in intracellular GSH level. PMID- 11379764 TI - NQ-Y15 inhibits the calcium mobilization by elevation of cyclic AMP in rat platelets. AB - 2-1(4-Cyanophenyl)aminol-3-chloro-1,4-naphthalenedione (NQ-Y15) is a dual action drug which acts as a thromboxane A2 (TXA2) synthase inhibitor and TXA2/PGH2 receptor antagonist. In the present study, we examined the effects of NQ-Y15 on Ca2+ mobilization, which is the common event in various types of platelet activation, in arachidonic acid (AA)-stimulated rat platelets. The elevation of cytosolic Ca2+ concentration ([Ca2+]i) induced by AA was inhibited by NQ-Y15 in a concentration-dependent manner. This inhibition-effect of NQ-Y15 was found to be based on the suppression of the rise in [Ca2+]i by the inhibition of both Ca2+ release from internal stores and Ca2+ influx from the extracellular space. Our successive trial was focused on the role of cyclic AMP (cAMP) in the action of NQ Y15, because cAMP was reported to be increased by dual action drugs such as picotamide and to inhibit the increase in [Ca2+]i. NQ-Y15 was confirmed to increase cAMP in AA-stimulated rat platelets. These results suggested that NQ-Y15 might inhibit the rise in [Ca2+]i in AA-treated rat platelets by increasing cAMP, which is involved in the inhibition of platelet activation. PMID- 11379765 TI - Antidiabetic effect of glycyrrhizin in genetically diabetic KK-Ay mice. AB - We, previously demonstrated that one shot administration of glycyrrhizin (Grz) reduced the postprandial blood glucose rise, using Std ddY mice. Subsequently, we evaluated the effects of long-term Grz treatment (2.7, 4.1 g/kg diet) on diabetic symptoms using genetically non-insulin dependent diabetic model mice (KK-Ay). Male KK-Ay mice were divided into 3 groups: the control group, 0.27% Grz diet (2.7 g of Grz/kg diet) group and 0.41% Grz diet (4.1 g of Grz/kg diet) group. The elevation of blood glucose concentration was almost entirely suppressed in mice fed the 0.41% Grz diet 7 weeks after the beginning of test feeding, although it was not suppressed in mice fed the control diet or the 0.27% Grz diet. Water intake in the control and 0.27% Grz diet groups increased gradually, whereas, this was not true in the 0.41% Grz diet group. Grz treatment significantly lowered blood insulin level. Throughout the experiment, Grz did not affect the food intake or body weight among the three groups. The mice fed the 0.41% Grz diet also improved their tolerance to oral glucose loading 9 weeks after the beginning of test feeding. This study shows that Grz has an antidiabetic effect in noninsulin-dependent diabetes model mice. PMID- 11379766 TI - Evaluation of the effects of restraint and footshock stress on small intestinal motility by an improved method using a radionuclide, 51Cr, in the rat. AB - The effect of two different stress stimuli, restraint stress and footshock stress, on small intestinal motility was evaluated by a more reliable method with improvement of the previous method using a radionuclide, 51Cr. The small intestinal transit was significantly inhibited by restraint stress, but not by footshock stress, although plasma corticosterone levels were significantly elevated to the same extent by restraint stress and footshock stress. These results suggest that restraint stress and footshock stress stimuli influence small intestinal motility via different mechanisms, but the reason for the difference is unclear. This experimental system using 51Cr seems to be useful for the elucidation of mechanisms for restraint stress-induced dysfunction of small intestinal motility because of its excellent quantitative evaluation of small intestinal transit. PMID- 11379767 TI - Muscarinic receptor binding characteristics in rat tissues after oral administration of oxybutynin and propiverine. AB - Ex vivo muscarinic receptor binding of oxybutynin and propiverine, the most commonly used anticholinergic agents for the treatment in patients with urinary incontinence, was investigated in rat tissues. The oral administration of oxybutynin (50.8 and 127 micromol/kg) caused a significant increase in the apparent dissociation constant (Kd) for specific (-)-[3H]QNB binding in the rat bladder, prostate, submaxillary gland, heart and cerebral cortex, compared with each of the control values. Also, in the submaxillary gland of these rats, there was a reduction in the maximal number of binding sites (Bmax) for (-)-[3H]QNB binding. Similarly, oral administration of propiverine at doses of 74.3-297 micromol/kg brought about a significant increase in the Kd values for (-)-[3H]QNB binding in rat tissues including the bladder, and greater increase in Kd values was seen in the rat prostate, heart and submaxillary gland. On the other hand, oral administration of propiverine, unlike oxybutynin, resulted in very little reduction in the Bmax valules for (-)-[3H]QNB binding in the submaxillary gland. In conclusion, the present study has shown that oxybutynin and propiverine, after oral administration, bind significantly to muscarinic receptors in tissues such as the bladder, which is the target organ for the treatment of urinary incontinence, and that oxybutynin appears to exhibit long-term binding to muscarinic receptors in the salivary gland. PMID- 11379768 TI - Ameliorative effects of paeoniflorin, a major constituent of peony root, on adenosine A1 receptor-mediated impairment of passive avoidance performance and long-term potentiation in the hippocampus. AB - We examined the effects of paeoniflorin on adenosine A1 receptor-mediated memory disturbance in the mouse passive avoidance test and inhibition of long-term potentiation (LTP) in the rat hippocampal CA1 region. The pretraining administration of the selective adenosine A1 receptor agonist N6 cyclopentyladenosine (CPA) significantly impaired the retention performance determined 24 h after the training test. The intraperitoneal injections of paeoniflorin and the adenosine A1 receptor antagonist 1,3-dipropyl-8 cyclopentylxanthine (DPCPX) significantly attenuated the deficit in retention performance caused by CPA. The in vitro studies revealed that adenosine (1 and 10 microM) dose dependently reduced both the population spike (PS) amplitudes and the tetanic stimulation-induced LTP in the hippocampus. DPCPX, at the concentration (0.1 microM) that had no effect on PS amplitudes or LTP induction, significantly reversed the suppressive effects of adenosine on both indices. Paeoniflorin also dose dependently reversed 10 microM adenosine-induced suppression of LTP but had no effect on PS reduced by adenosine. These results suggest that paeoniflorin ameliorates memory disruption mediated by adenosine A1 receptor and that modulation of adenosine-mediated inhibition of LTP in the hippocampus is implicated in its beneficial effect on learning and memory impairment in rodents. PMID- 11379769 TI - Inhibitory effects of TA-993 and its metabolite MB3 on platelet activation induced by collagen and U-46619 in human platelets. AB - In the present study, we investigated the effects of TA-993 and its metabolite MB3 on platelet activation in vitro. TA-993 and MB3 concentration-dependently inhibited platelet aggregation and ATP release induced by collagen in human platelets. Thromboxane (Tx) A2 formation, as determined by the production of TxB2, and the increase in intracellular Ca2+ concentration ([Ca2+]i) were also suppressed by TA-993 and MB3. TA-993 and MB3 did not inhibit TxA2 formation caused by arachidonic acid. These results suggest that the inhibition of platelet activation by TA-993 and MB3 is partly mediated by an inhibition of TxA2 formation at a step prior to cyclooxygenase. Furthermore, TA-993 and MB3 inhibited U-46619-induced platelet aggregation without blockade of the increase in [Ca2+]i, suggesting that they are likely to exert some additional effects on the intracellular events induced by Ca2+. PMID- 11379770 TI - Effect of 4-(4-chlorobenzyl)pyridine on rat hepatic microsomal cytochrome P450 and drug-metabolizing enzymes in vivo and in vitro. AB - The effect of 4-(4-chlorobenzyl)pyridine (4-CBP) on rat hepatic microsomal cytochrome P450 (P450) and its molecular species (CYP2B1, 2E1, 3A2, 2C11, and 2C12), and on drug-metabolizing enzyme activities were examined in vivo and in vitro. Treatment of rats with 4-CBP resulted in the induction of P450 and drug metabolizing enzymes in a dose-dependent manner, but it was markedly inhibitory at higher dose levels. Immunoblot analyses revealed that 4-CBP induces both CYP2B1 and 2E1; however, both were decreased by increasing the dose of 4-CBP. The in vitro inhibitory experiment revealed that 4-CBP strongly inhibited benzphetamine N-demethylase activity, but not dimethylnitrosamine N-demethylase activity. The present findings provide information on the induction and inhibition effect of chlorinated benzylpyridine on hepatic microsomal P450s and drug-metabolizing enzymes in vivo and in vitro. PMID- 11379771 TI - Oral administration of dimethylarsinic acid, a main metabolite of inorganic arsenic, in mice promotes skin tumorigenesis initiated by dimethylbenz(a)anthracene with or without ultraviolet B as a promoter. AB - Concerning arsenic-induced tumorigenesis, an animal model must be developed for understanding the mechanism of human carcinogenesis by arsenics. To determine whether orally administered dimethylarsinic acid (DMA) promotes and causes the progression of skin tumorigenesis, an animal protocol by topical application of dimethylbenz(a)anthracene (DMBA) with or without UVB, a tumor promoter, in hairless mice was used. The administration of DMA by the oral route promoted not only the formation of papillomas induced by DMBA alone but also the formation of malignant tumors induced by way of the formation of atypical keratoses by treatment with DMBA and UVB. A phenomenon, the progression of keratoses- >atypical keratoses-->squamous cell carcinomas (SCCs), observed in the present study may resemble the development of tumors in arsenic-exposed humans. We also discussed the involvement of a reactive oxygen species (ROS), e.g., the dimethylarsenic peroxy radical [(CH3)2AsOO.], produced during the metabolic processing of DMA, in skin and in multi-organ tumorigenesis. PMID- 11379772 TI - Cytotoxicity of reactive oxygen species and related agents toward undifferentiated and differentiated rat phenochromocytoma PC12 cells. AB - The cytotoxicity of reactive oxygen species and related agents toward cultured rat adrenal medullary phenochromocytoma PC12 cells was examined. These species and agents include hydrogen peroxide, linoleic acid hydroperoxide (LOOH), tert butyl hydroperoxide, paraquat, 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH), 2,2'-azobis(2,4-dimethylvaleronitrile) (AMVN), and a hypoxanthine xanthine oxidase system. The respective 50% lethal concentrations (LC50) for undifferentiated and differentiated PC12 cells were found to be 275 and 165 microM of hydrogen peroxide, 58.3 and 35.3 microM of LOOH, 536 and 212 microM of tert-butyl hydroperoxide, 42.5 and 26.5 mM of paraquat, 79.5 and 74.5 mM of AAPH, 412 and 300 microM of AMVN, and 37.2 and 16.6mU x ml(-1) xanthine oxidase activity of the hypoxanthine-xanthine oxidase system. These results show that the differentiated cells were more susceptible to these oxidative agents than the undifferentiated cells. The glutathione peroxidase activity level of the undifferentiated cells was 2-3 times higher than the differentiated cells, the catalase activity level also tended to be higher, the superoxide dismutase activity level was higher on a per-protein-quantity basis but lower on a per-cell number basis, and the total and reduced glutathione concentration levels were considerably higher. The enhanced susceptibility of the differentiated cells may result from decreases in the activity of glutathione peroxidase and the concentration of its substrate, reduced glutathione (GSH). Further, the preincubation of PC12 cells with alpha-tocopherol or L-buthionine-(R,S) sulfoximine (BSO) lowered or enhanced their cytotoxicities, respectively. PMID- 11379773 TI - Isolation of plasma kallikrein by high efficiency affinity chromatography and its characterization. AB - We established a simple method for the purification of human plasma kallikrein (PK) by affinity chromatography and characterized it by analytical reverse phase HPLC and Time of Flight Mass Spectroscopy (TOF-MS). The affinity resin (PKSI Toyopearl) was synthesized using a selective synthetic inhibitor of plasma kallikrein (PKSI-527) as a ligand. The resin was found to have the highest efficiency in PK purification when the coupling ratio of PKSI-527 per resin was 9 14 micromol/g. PK was purified 466-fold with a yield of 83% from acetone activated human plasma by affinity chromatography. The purity of PK thus obtained was confirmed by reverse phase-HPLC with a linear gradient of acetonitrile. The molecular weight of the purified PK was determined to be 86,151 by TOF-MS. PMID- 11379774 TI - Antiproliferative activity of diarylheptanoids from the seeds of Alpinia blepharocalyx. AB - The 95% EtOH extract of the seeds of Alpinia blepharocalyx (Zingiberaceae) showed significant antiproliferative activity towards human HT-1080 fibrosarcoma and murine colon 26-L5 carcinoma cells. Chemical investigation of the extract led to the isolation of forty-four new (1-44) and one known (45) diarylheptanoids, eleven phenolic compounds (46-56) together with beta-sitosterol glucoside (57). Almost all the isolated compounds showed significant antiproliferative activity in a concentration-dependent manner. Among the compounds, epicalyxin F (17) exhibited the most potent activity against the proliferation of colon 26-L5 carcinoma cells with an ED50 value of 0.89 microM, while calyxin B (2) exhibited the most potent activity against human HT-1080 fibrosarcoma cells with an ED50 value of 0.69 microM. Moreover, calyxins B (2) and K (11), epicalyxins F (17), I (20) and K (22), 6-hydroxycalyxin F (25), blepharocalyxin B (27) and mixtures of 7 and epicalyxin G (18) and of calyxin J (10) and epicalyxin J (21) possessed more potent activity than a clinically used anticancer drug, 5-fluorouracil, towards HT-1080 fibrosarcoma cells. Analysis of the structure activity relationship suggested that the position of the attachment of a chalcone or a flavanone moiety does not affect the activity, although their presence in association causes a substantial enhancement of the antiproliferative activity. Moreover, the conjugated double bond of the chalcone moiety and the phenolic hydroxyl group potentiate the antiproliferative activity of the compounds. PMID- 11379775 TI - Effects of 4-hydroxyantipyrine and its 4-O-sulfate on antipyrine as biodistribution promoter. AB - The effects of 4-hydroxyantipyrine (4-OH), a major metabolite of antipyrine, and its 4-O-sulfate (4-S) on the pharmacokinetics of antipyrine were investigated in rats. Plasma elimination of intravenously administered antipyrine was significantly decelerated under a steady-state concentration of 4-OH but not under that of 4-S. Tissue-to-plasma concentration ratio (Kp) of antipyrine under its steady-state concentration was significantly increased in the brain and heart by the concomitant use of 4-OH, while similar use of 4-S had no effect. The enhancement of the blood-brain barrier (BBB) permeability of antipyrine caused by the concomitant use of 4-OH was believed to be concerned with the increase of the Kp value of antipyrine in the brain. These results suggested that 4-OH could be used as a biodistribution promoter. PMID- 11379776 TI - Characteristics of tissue distribution of various polysaccharides as drug carriers: influences of molecular weight and anionic charge on tumor targeting. AB - Using the Walker 256 model for carcinosarcoma-bearing rats, we intravenously administered 5 polysaccharide carriers with various molecular weights (MWs) and electric charges and tested for their plasma and tissue distribution. Two carriers, carboxymethylated-D-manno-D-glucan (CMMG) and CMdextran (CMDex), showed higher plasma AUC than the other carriers tested, namely, CMchitin (CMCh), N desulfated N-acetylated heparin (DSH), and hyaluronic acid (HA). This was consistently found to be true over the range of MWs tested. For CMDex, the maximum value of plasma AUC was obtained when the MW exceeded 150 kDa. As for the anionic charge, CMDex (110-180 kDa) with a degree of substitution (DS) of the CM groups ranging from 0.2 to 0.6, showed maximum plasma AUC values. Twenty-four hours after administration, the concentration of CMDex (180-250 kDa; DS: 0.6-1.2) in tumors was more than 3% of dose/g--approximately 10-fold higher than those observed with CMCh, DSH and HA. Doxorubicin (DXR) was bound to these carriers via a peptide spacer, GlyGlyPheGly (GGFG), to give carrier-GGFG-DXR conjugates (DXR content: 4.2-7.0 (w/w)%), and the antitumor effects of these conjugates were tested with Walker 256 carcinosarcoma-bearing rats by monitoring the tumor weights after a single intravenous injection. Compared with free DXR, CMDex-GGFG DXR and CMMG-GGFG-DXR conjugates significantly suppressed tumor growth, while the CMCh-GGFG-DXR, DSH-GGFG-DXR, and HA-GGFG-DXR conjugates in a similar comparison showed weak tumor growth inhibition. These findings suggest that the antitumor effect of the carrier-DXR conjugates was related to the extent with which the carriers accumulated in the tumors. PMID- 11379777 TI - N-Acetyltransferase2 genotype correlated with isoniazid acetylation in Japanese tuberculous patients. AB - Isoniazid (INH) is metabolized by polymorphic N-acetyltransferase2 (NAT2). In the present study, the relationship between the NAT2 genotype and the INH acetylator phenotype was examined in Japanese tuberculous patients and compared with healthy subjects. Subjects were classified according to the genotyping into NAT2*5B (allele4), NAT2*6A (allele3) and NAT2*7B (allele2), using the PCR-RFLP method. Twelve healthy subjects and 7 tuberculous patients participated in the INH acetylator phenotyping study, in which each subject was administered an oral dose of INH, followed by urine sampling for 24 h. Urinary concentrations of INH and N acetylisoniazid (AcINH) were measured by the HPLC method. The urinary recoveries of INH (% of dose) in healthy subjects in relation to NAT2 genotyping were as follows: 6.4+/-2.2 in the homozygotes for the wild-type allele, 10.7+/-2.2 in the compound heterozygotes for the mutant allele, and 38.6+/-6.4 in the homozygotes for the mutant allele. In the patients study, the findings in the corresponding three groups were 4.0+/-1.7, 8.8 and 18.3+/-9.3. Although no significant difference was found because of the lower systemic exposure of INH in patients compared with healthy subjects, there were differences in the disposition kinetics of INH between subjects with and without mutations in the NAT2 gene, and these findings were observed not only in healthy subjects but also in patients who had comedicated drugs and hepatic dysfunctions. The findings indicated that the metabolism of INH by NAT2 is clearly impaired in subjects with mutations in the NAT2 gene, and thus genotyping for three NAT2 point mutations was adequate to predict the metabolism of INH in Japanese tuberculous patients as well as healthy subjects. This NAT2 genotyping could become a useful alternative to TDM for INH. PMID- 11379778 TI - A comparative pharmacodynamic study of the arrhythmogenicity of antidepressants, fluvoxamine and imipramine, in guinea pigs. AB - Among several classes of antidepressants, tricyclic antidepressants are known to prolong QTc intervals (QT interval corrected by heart rate) in electrocardiograms, while selective serotonin uptake inhibitors (SSRI) are considered to be devoid of arrhythmogenicity. In this study, we aimed to compare the arrhythmogenic potencies of imipramine (IMI), a typical tricyclic antidepressant, and fluvoxamine (FLV), an SSRI, at therapeutic and supratherapeutic concentrations using guinea pigs in vivo. Guinea pigs were anesthetized, and IMI (10 and 20 mg/kg/h) or FLV (20 mg/kg/h) was intravenously administered for 90 minutes to obtain the time-courses of drug concentrations in plasma and the changes in the QTc intervals during and after the drug administration. IMI induced distinct QTc prolongation in a dose-dependent manner, while FLV prolonged QTc intervals only slightly. A pharmacokinetic pharmacodynamic analysis revealed that the potency for QTc prolongation of IMI was 1.7-fold higher than that of FLV. Taking the therapeutic concentration into account, the clinical risk of FLV for QTc prolongation was suggested to be 5-fold lower than that of IMI. Therefore, this SSRI agent was suggested to be safer than the tricyclic antidepressant for patients with cardiac risk factors, including arrhythmia, or for those taking other arrhythmogenic drugs concomitantly. PMID- 11379779 TI - Increase in doxorubicin cytotoxicity by inhibition of P-glycoprotein activity with lomerizine. AB - Acquired resistance to chemotherapy is a major problem during cancer treatment. One mechanism for drug resistance is overexpression of the MDR (multidrug resistance)1 gene encoding the transmembrane efflux pump, P-glycoprotein (P-gp). Calcium channel blockers such as verapamil, nifedipine and nicardipine have been shown to reverse cellular drug resistance by inhibiting P-gp drug efflux. This study evaluated whether a new calcium channel blocker, lomerizine, influenced doxorubicin (Dox) cytotoxicity and P-gp activity in a P-gp-expressing cell line compared to a non-expressing subline. Verapamil, and even more markedly, lomerizine, increased cellular uptake of calcein transported by P-gp in a P-gp expressing erythroleukemia cell line, K562-Dox. Ten microM of lomerizine reduced the IC50 of doxorubicin in the K562-Dox from 60000 ng/ml to 800 ng/ml, whereas the IC50 of doxorubicin in the K562 subline was only marginally affected by these drugs. Lomerizine showed greater reduction in P-gp efflux than verapamil at an equimolar concentration. These results suggest that lomerizine has the clinical potential to reverse tumor MDR involving the efflux protein P-gp. PMID- 11379780 TI - Antioxidant potential of qizhu tang, a chinese herbal medicine, and the effect on cerebral oxidative damage after ischemia reperfusion in rats. AB - A traditional Chinese herbal medicine, Qizhu Tang (QZT) was studied for its in vitro antioxidant activity and the effect on cerebral oxidative damage after forebrain ischemia followed by reperfusion in rats. The QZT decoction was shown to have strong hydroxyl radical (*OH) scavenging activity (approx. 0.1 mM as Trolox equivalent) when determined by ESR using DMPO as a spin trap reagent and H2O2/UV as the *OH source. When the QZT decoction was injected into rat duodenum 2 h before cerebral ischemia, the oxidative brain damage after 45 min reperfusion was strongly inhibited in terms of two biochemical indications, thiobarbituric acid reactive substance formation and the loss of glutathione peroxidase. Since the QZT formula consists of 4 herbal constituents (Rhizoma atractylodis, Poria, Radix notoginseng and Radix astragali), each of the component herbs and their combinations were also examined for their protective effects on the cerebral ischemia/reperfusion injury and the effects were compared with their in vitro antioxidant potential. Although some of the incomplete formulas showed as strong antioxidant activities as complete QZT in vitro, only the complete QZT formula was effective in preventing cerebral oxidative injury in rats, and other preparations showed limited activity in vivo. PMID- 11379781 TI - Gas chromatography-mass spectrometric determination of activity of human placental aromatase using 16alpha-hydroxyandrostenedione as a substrate. AB - Aromatization of 16alpha-hydroxyandrostenedione (16alpha-OH AD) with aromatase in human placental microsomes was studied by gas chromatography-mass spectrometry (GC-MS) using 12,4,6,6,9alpha,16beta,17alpha-2H7]estriol as an internal standard. 16alpha-OH AD was incubated with the microsomes in the presence of NADPH in air. The metabolite was extracted with ethyl acetate and treated with NaBH4. The reduced product, estriol, was isolated by Sep-Pak C18 cartridge and then analyzed as the tris(trimethylsilyl)ether by a GC-MS (EI mode). The production of estriol was dependent upon protein concentration and incubation time. Apparent Km and Vmax values of the microsomal aromatase for 16alpha-OH AD were 568 nM and 25.5 pmol/min/mg protein, respectively. In this assay, aromatase activity, estriol formation, could be determined at a level as low as 1 pmol/min/mg protein. Aromatase inhibitors, 4-hydroxy- and 6-oxo-androstenediones, prevented the estriol formation in a competitive manner with 25 and 30 nM of apparent Ki values, respectively. PMID- 11379782 TI - A sensitive assay of human blood platelet cyclic nucleotide phosphodiesterase activity by HPLC using fluorescence derivatization and its application to assessment of cyclic nucleotide phosphodiesterase inhibitors. AB - A selective and sensitive HPLC measurement of 3',5'-cyclic nucleotide phosphodiesterase (PDE) activity in human platelets using (3,4 dimethoxyphenyl)glyoxal (DMPG) as a fluorogenic reagent for guanine and its nucleosides and nucleotides is described. cGMP, a substrate for PDE, and GMP, which was produced by the enzyme reaction, are selectively converted by the reaction with DMPG to the fluorescent derivatives. The derivatives were separated by reversed-phase HPLC. Human platelet PDE activity was measured and the inhibitory effects of several compounds were investigated. PMID- 11379783 TI - Cloning and sequencing of the astA gene encoding arylsulfate sulfotransferase from Salmonella typhimurium. AB - Arylsulfate sulfotransferase (ASST) transfers a sulfate group from a phenolic sulfate ester to a phenolic acceptor substrate. In the present study, the gene encoding ASST was cloned from a genomic library of Salmonella typhimurium. The gene was subcloned into the vector pKF3 and was sequenced. A recombinant clone harboring the gene was directly identified using a fluorescent assay. Sequencing revealed two contiguous open reading frames (ORFs) on the same strand. Based on amino acid sequence homology, ORF1 and ORF2 are designated as astA and dsbA, respectively. The deduced amino acid sequence of astA from S. typhimurium was highly similar to those of the Enterobacter amnigenus, Klebsiella, and Campylobacter jejuni ASSTs, encoded by the astA genes. However, an ASST activity assay revealed a different acceptor specificity. Using p-nitrophenyl sulfate (PNS) as a donor substrate, phenol is the best acceptor substrate, followed by alpha-naphthol, resorcinol, tyramine, acetaminophen, and tyrosine. PMID- 11379784 TI - Water-soluble antioxidants inhibit macrophage recognition of oxidized erythrocytes. AB - Effect of exogenously added water-soluble antioxidants on the mouse macrophage lectin-like receptor activity for oxidized erythrocytes was investigated. A monolayer of thioglycollate-induced mouse peritoneal macrophages was preincubated with each of the antioxidants at 37 degrees C for 1 h, and the binding for mouse erythrocytes oxidized with ADP-chelated Fe(III) was examined. The binding was decreased by preincubation of macrophages with ascorbic acid-related compounds including ascorbic acid, erythorbic acid and dehydroascorbic acid in a dose dependent fashion at relatively high concentrations above 10 microM. The binding was similarly decreased by preincubation of macrophages with catechin compounds including epicatechin, epigallocatechin, epicatechin gallate and epigallocatechin gallate in a dose-dependent fashion at 0.01-100 microM. The binding was more effectively decreased by preincubation of macrophages with thiol-related compounds including glutathione, oxidized glutathione, glutathione isopropyl ester and N-acetylcysteine in a dose dependent fashion at relatively low doses below 1 microM. These results showed that water-soluble antioxidants especially glutathione and its derivatives reduced the ability of macrophages to bind oxidized erythrocytes, suggesting that the activity of lectin-like receptors of macrophages for oxidized erythrocytes was regulated by oxidative mechanisms. PMID- 11379785 TI - The relationship between plasma high density lipoprotein cholesterol levels and cholesteryl ester transfer protein activity in six species of healthy experimental animals. AB - Plasma high-density lipoprotein cholesterol (HDL-C) concentrations are regulated by plasma cholesteryl ester transfer protein (CETP) in humans. The aim of this study was to ascertain the relationship between plasma HDL-C and plasma CETP activities in mouse, rat, dog, hamster, rabbit and monkey. In this study, the plasma HDL-C levels were highest in dogs and lowest in rabbits among the six species. Plasma CETP activities were higher in hamsters, rabbits and monkeys compared to mice, rats and dogs. The present study shows that there are species differences in HDL-C and CETP activity in six species of healthy experimental animals, with the six species being separated into two types. The first type showed a high HDL-C/TC ratio with low or absent CETP activity, and included mouse, rat and dog, whereas the second type showed a low HDL-C/TC ratio and high CETP activity, and included hamster, rabbit and monkey. The present study also shows that there is a strong relationship between plasma HDL-C levels and CETP activity in high CETP activity animals and that the relationship between the HDL C/TC ratio and CETP activity is an important factor in all animals, regardless of CETP activity level. PMID- 11379786 TI - Metabolism of chiisanoside from Acanthopanax divaricatus var. albeofructus by human intestinal bacteria and its relation to some biological activities. AB - The metabolic pathway of chiisanoside isolated from leaves of Acanthopanax divaricatus var. albeofructus (Araliaceae) by human intestinal bacteria and by the protein fraction of leaves of this plant were investigated, and the cytotoxic and anti-rotaviral activities of chiisanoside and its metabolite, chiisanogenin, were assayed. Chiisanogenin was produced as a main metabolite, when chiisanoside were incubated for 15 h with human intestinal bacteria. This metabolic pathway proceeded more potently with the protein fraction than with human intestinal bacteria. The in vitro cytotoxicity of chiisanogenin was superior to that of chiisanoside. H+/K+ ATPase was more potently inhibited by chiisanogenin than by chiisanoside. However, the anti-rotaviral activity of chiisanoside was more potent than that of chiisanogenin. PMID- 11379787 TI - Testosterone 5alpha-reductase inhibitory active constituents from Anemarrhenae Rhizoma. AB - The diethyl ether extract of Anemarrhenae Rhizoma (rhizomes of Anemarrhena asphodeloides Bunge) showed testosterone 5alpha-reductase inhibitory activity. Two major constituents, cis-hinokiresinol (1) and 2,6,4'-trihydroxy-4 methoxybenzophenone (2) were identified as the active principles. The inhibitory activity of 1 was superior to that of ethinylestradiol, but that of 2 was weak. PMID- 11379788 TI - Transdermal absorption of bupranolol in rabbit skin in vitro and in vivo. AB - This study was designed to clarify the percutaneous penetration of bupranolol (BP), a beta-adrenoceptor antagonist, through rabbit skin and to compare the in vitro penetration with the in vivo absorption. BP penetrated across the skin slowly in the absence of enhancers in vitro. Isopropyl myristate and N-methyl-2 pyrrolidone enhanced the in vitro penetration, with a 3.6 times higher flux compared with that without enhancers. However, in the in vivo percutaneous absorption, the maximal penetration was obtained with the formulation added dlimonene, with a 3.0 times higher area under the concentration-time curve (AUC) than that for the formulation without enhancers. The plasma levels of BP determined, however, were extremely lower than the theoretical plasma steady state concentrations predicted. The plasma levels of BP after application of these formulations were maintained in the range of 7-22 ng/ml for 30 h, of which concentrations were above the therapeutically effective concentration (1.5-4 ng/ml). Therefore, the transdermal systems will offer an efficient drug delivery system for the treatment of angina pectoris and tachycardia. PMID- 11379789 TI - Effects of leucocyanidines on activities of metabolizing enzymes and antioxidant enzymes. AB - Procyanidolic oligomers (leucocyanidines, LCs) extracted from grape seeds (Vitis vinifera) are known to have antioxidant and antimutagenic activities, and a protective effect against cardiovascular disease. In the present study we examined the influence of LCs on the activities of phase 1 enzymes and conjugation enzymes and on antioxidant enzymes such as superoxide dismutase, catalase, and glutathione peroxidase. Administration of LCs (25, 50, and 100 mg/kg. p.o. for 7 d) markedly decreased the activities of NADPH-cytochrome P450 reductase, P4501A1, P4501A2, and P4503A4, but significantly increased the activities of glutathione S-transferase and phenolsulfotransferase in rat liver. However, the activities of antioxidant enzymes were not affected by LC administration. The inhibition of P450s and increases in phase II enzyme activities indicate a role for LCs as a chemopreventive agent against toxic or carcinogenic metabolites of P450 isozymes. PMID- 11379790 TI - Brain endothelial adhesion molecule expression in experimental colitis. AB - OBJECTIVES: 1) To determine if endothelial expression of adhesion molecules involved in leukocyte recruitment is increased in the brain and other organs in four different models of experimental colitis, and 2) to investigate whether leukocyte infiltration occurs in the brain of colitic animals. METHODS: Endothelial vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) expression was quantified, using the dual radiolabeled antibody technique in rats with trinitrobenzenesulfonic acid (TNBS)-induced colitis, in mice with dextran sulfate sodium (DSS)-induced colitis, in SCID mice reconstituted with CD45RBhigh T-cells, and in IL-10-/- mice. Leukocyte infiltration in the brain of TNBS-induced colitic rats was assessed by myeloperoxidase activity and immunohistochemical staining with anti-CD45 monoclonal antibody. RESULTS: Marked upregulation of brain endothelial VCAM-1 (2- to 5.5-fold) was consistently found in colitic animals in the four models studied. Brain VCAM-1 strongly correlated with colon VCAM-1 and colon weight. By contrast, upregulation of brain ICAM-1 in colitic animals was only observed in the CD45RBhigh transfer (3-fold) and the TNBS-induced (1.5-fold models). Heart and muscle VCAM-1 and ICAM-1 were not upregulated in colitic animals in the majority of models studied. There was no leukocyte infiltration into the brain of TNBS-induced colitic rats. CONCLUSIONS: Our study demonstrates a marked and specific upregulation of endothelial VCAM-1 in the brain of colitic animals. This activation of cerebral endothelial cells was not associated with an infiltration of leukocytes into brain tissue. PMID- 11379791 TI - Altered mechanisms underlying hypoxic dilation of skeletal muscle resistance arteries of hypertensive versus normotensive Dahl rats. AB - OBJECTIVE: To determine mechanisms underlying hypoxic dilation of skeletal muscle resistance arteries from normotensive (NT) and hypertensive (HT) Dahl salt sensitive (SS) rats. METHODS: Isolated gracilis arteries (GA) from both rat groups were viewed via television microscopy and vascular responses to a reduction in PO2 from 145 mm Hg to 40 mm Hg were measured with a video micrometer. Responses were determined following endothelium removal and following inhibition of specific biochemical pathways regulating vascular tone. RESULTS: Hypoxic dilation was impaired in HT rats versus NT controls. Endothelium removal abolished hypoxic dilation in NT rats, although a significant dilation to hypoxia remained in vessels from HT animals. Inhibition of cytochrome P450 (CP450) 4A enzymes blunted hypoxic dilation in both groups, while inhibition of epoxyeicosatrienoic acid (EET) production impaired responses in NT rats only. Inhibition of 20-hydroxyeicosatetraenoic acid (20-HETE) production or blockade of membrane receptors for 20-HETE reduced hypoxic dilation in HT rats, with minimal effects in NT animals. Nitric oxide synthase inhibition had no effect on hypoxic dilation in either group, while cyclooxygenase inhibition significantly reduced this response in both groups. CONCLUSIONS: These results suggest that the mechanisms of hypoxic dilation in GA from NT Dahl-SS rats are altered with HT, impairing the response to reduced PO2. While hypoxia induces substantial prostanoid release in both groups, the role of CP450 4A enzymes is shifted from EET production in NT rats toward inhibition of 20-HETE production in HT rats. PMID- 11379792 TI - Microvascular endothelial cell death and rarefaction in the glucocorticoid induced hypertensive rat. AB - OBJECTIVE: The objective was to quantify and to describe microvascular endothelial cell death in glucocorticoid-induced hypertension. Microvascular rarefaction, which has been shown in human and animal hypertension, may result from increased endothelial cell apoptosis. METHODS: Wistar rats were administered the synthetic glucocorticoid dexamethasone (0.5 mg/kg b.w. per day, i.m.) for 5 days and were compared with a group of control rats treated with the vehicle. In vivo microvascular endothelial cell death was quantified by propidium iodide fluorescent labeling in the mesentery. Normal nuclear DNA (labeled with Hoechst 33342) and DNA with apoptotic characteristics in muscle endothelium (labeled with ethidium bromide) were visualized using confocal imaging. Microvessel length density was measured by using a fluorescein isothyocyanate-labeled lectin technique. RESULTS: The dexamethasone-treated rats exhibit approximately a 10% increase in cell death along the mesenteric arteriolar and venular endothelium compared with controls. Confocal analysis of cremaster muscle in dexamethasone treated rats demonstrated nuclear fragmentation and decreased nuclear volumes in dying endothelial cells, which are consistent with an apoptotic process. The capillary length density in cremaster muscle was decreased on average by 16% in the hypertensive rats. CONCLUSIONS: These results suggest capillary structural rarefaction by an increased rate of apoptotic endothelial cell death in glucocorticoid-induced hypertension. PMID- 11379793 TI - Mechanisms of leukocyte sequestration in inflamed lungs. AB - The lungs are an important site of host defense. The capillary blood contains an increased concentration of neutrophils and other leukocytes compared with large vessels, due to the structure of the pulmonary capillary bed, the diameter of spherical leukocytes, and their poor deformability compared with erythrocytes. During inflammation within the distal airways, neutrophils sequester within the pulmonary capillaries and emigrate into the parenchyma. This sequential process involves complex events regulating interactions between mechanical and adhesive properties of both neutrophils and endothelial cells. Initial changes in the cytoskeleton may stiffen the neutrophils and prevent them from deforming, while subsequent dynamic cytoskeletal remodeling of neutrophils and endothelial cells results in crawling and transendothelial migration. Emigration of neutrophils can occur through at least two adhesion pathways: one that requires the CD11/CD18 adhesion complex and one that does not. Which pathway is selected is determined by the stimulus and the signaling pathways that are initiated. Migration through the alveolo-capillary wall is also highly regulated. Neutrophils released from the bone marrow traffic first through the pulmonary microvasculature, and the phenotype of these newly released neutrophils impacts pulmonary host defense. The many recent studies underline the complexity of neutrophil responses and host defense and the uncertainties of our knowledge. PMID- 11379794 TI - Regulation of blood-brain barrier permeability. AB - The blood-brain barrier minimizes the entry of molecules into brain tissue. This restriction arises by the presence of tight junctions (zonulae occludens) between adjacent endothelial cells and a relative paucity of pinocytotic vesicles within endothelium of cerebral arterioles, capillaries, and venules. Many types of stimuli can alter the permeability characteristics of the blood-brain barrier. Acute increases in arterial blood pressure beyond the autoregulatory capacity of cerebral blood vessels, application of hyperosmolar solutions, application of various inflammatory mediators known to be elevated during brain injury, and/or activation of blood-borne elements such as leukocytes can produce changes in permeability of the blood-brain barrier. The second messenger systems that account for increases in permeability of the blood-brain barrier during pathophysiologic conditions, however, remain poorly defined. This review will summarize studies that have examined factors that influence disruption of the blood-brain barrier, and will discuss the contribution of various cellular second messenger pathways in disruption of the blood-brain barrier during pathophysiologic conditions. PMID- 11379795 TI - Regulation of neurotrophin signaling in aging sensory and motoneurons: dissipation of target support? AB - A hallmark of senescence is sensorimotor impairment, involving locomotion and postural control as well as fine-tuned movements. Sensory and motoneurons are not lost to any significant degree with advancing age, but do show characteristic changes in gene-expression pattern, morphology, and connectivity. This review covers recent experimental findings corroborating that alterations in trophic signaling may induce several of the phenotypic changes seen in primary sensory and motoneurons during aging. Furthermore, the data suggests that target failure, and/or breakdown of neuron-target interaction, is a critical event in the aging process of sensory and motoneurons. PMID- 11379799 TI - Measuring allergen exposure in the home: who benefits? PMID- 11379796 TI - The augmentation hypothesis for improvement of antidepressant therapy: is pindolol a suitable candidate for testing the ability of 5HT1A receptor antagonists to enhance SSRI efficacy and onset latency? AB - The development of selective serotonin reuptake inhibitors (SSRIs) provided a major advancement in the treatment of depression. However, these drugs suffer from a variety of drawbacks, most notably a delay in the onset of efficacy. One hypothesis suggests that this delay in efficacy is due to a paradoxical decrease in serotonergic (5-HT) neuronal impulse flow and release, following activation of inhibitory presynaptic 5-HT1A autoreceptors, following acute administration of SSRIs. According to the hypothesis, efficacy is seen only when this impulse flow is restored following desensitization of 5-HT1A autoreceptors and coincident increases in postsynaptic 5-HT levels are achieved. Clinical proof of this principal has been suggested in studies that found a significant augmenting effect when the beta-adrenergic/5-HT1A receptor antagonist, pindolol, was coadministered with SSRI treatment. In this article, we review preclinical electrophysiological and microdialysis studies that have examined this desensitization hypothesis. We further discuss clinical studies that utilized pindolol as a test of this hypothesis in depressed patients and examine preclinical studies that challenge the notion that the beneficial effect of pindolol is due to functional antagonism of the 5-HT1A autoreceptors. PMID- 11379797 TI - Serum growth factors and neuroprotective surveillance: focus on IGF-1. AB - The adult brain requires a constant trophic input for appropriate function. Although the main source of trophic factors for mature neurons is considered to arise locally from glial cells and synaptic partners, recent evidence suggests that hormonal-like influences from distant sources may also be important. These include not only relatively well-characterized steroid hormones that cross the brain barriers, but also blood-borne protein growth factors able to cross the barriers and exert unexpected, albeit specific, trophic actions in diverse brain areas. Insulin-like growth factor I (IGF-I) is until now the serum neurotrophic factor whose actions on the adult brain are best-characterized. This is because IGF-I has been known for many years to be present in serum, whereas the presence in the circulation of other more classical neurotrophic factors has only recently been recognized. Thus, new evidence strongly suggests that IGF-I, and other blood borne neurotrophic factors such as Fibroblast Growth Factor (FGF-2) or the neurotrophins, exert a tonic trophic input on brain cells, providing a mechanism for what we may refer to as neuroprotective surveillance. Protective surveillance includes "first-line" defense mechanisms ranging from blockade of neuronal death after a wide variety of cellular insults to upregulation of neurogenesis when defenses against neuronal death are overcome. Most importantly, surveillance should also encompass modulation of homeostatic mechanisms to prevent neuronal derangement. These will include modulation of basic cellular processes such as metabolic demands and maintainance of cell-membrane potential as well as more complex processes such as regulation of neuronal plasticity to keep neurons able to respond to constantly changing functional demands. PMID- 11379798 TI - Reconstructing smell. AB - Odorant signal transduction and neurogenesis are fundamental properties of the olfactory epithelium. Many preparations have been used to elucidate some of the mechanisms underlying these properties. In this article, we briefly review these research areas and describe some of the techniques used to obtain the data. We focus specifically on the cell-culture paradigm and the data obtained from various immortal cell lines in their attempts to reconstruct the olfactory epithelium in vitro. PMID- 11379800 TI - Asthma in the elderly: do they need an allergy evaluation? PMID- 11379801 TI - Update on nonallergic rhinitis. AB - BACKGROUND: Although nonallergic rhinitis is a well recognized entity, its incidence and therapy have not been definitively studied. Recent epidemiologic studies and treatment trials have furthered our knowledge regarding the frequency of occurrence of this disorder and effective treatment modalities. OBJECTIVE: To review and put into perspective recent advances in our knowledge regarding the incidence and significance as well as therapy of chronic nonallergic rhinitis. In addition, based upon these data, to propose a classification of this disorder. DATA SOURCES: The MEDLINE database and the results of a national survey of allergists (National Rhinitis Task Force) conducted in 15 allergy practices involving 975 patients. CONCLUSIONS: Nonallergic rhinitis is a common disease that probably affects as many as 17 million Americans. Of equal importance is that, based on available data, approximately 22 million people suffer with a combination of nonallergic rhinitis and allergic diseases (mixed rhinitis). Both nonallergic and mixed rhinitis occur more frequently in adults than in children, may be more common in female patients than in male patients, and are more likely to be perennial than seasonal. Agents demonstrating efficacy (based on controlled trials or having approval by the FDA) for the therapy of nonallergic rhinitis are azelastine and topical nasal steroids. PMID- 11379802 TI - Allergenic materials in the house dust of allergy clinic patients. AB - INTRODUCTION: Environmental agents including animal, fungal, tree, and weed antigens are known to cause allergic rhinitis and asthma. The following study was performed to measure the antigen concentration of several of these in house dust of children seen in an allergy clinic. Comparisons are made between household allergen levels of children seen for asthma and children seen for other reasons. METHODS: Dust samples were solicited from patients in a pediatric allergy specialty clinic and other individuals associated with the clinic. Persons submitting dust were asked to complete a questionnaire describing their house. Samples were extracted, centrifuged, and filtered for sterility. Samples were stored in 50% glycerol at -20 degrees C. Specific antigens for Alternaria, Cladosporium, Aspergillus, Candida, Dermatophagoides farinae, cat, dog, oak, fescue, ragweed, plantain, and cockroach were measured using inhibition assays developed with whole antigen extract. Allergens Der p1, Der f 1, Alt a 1, and Alt a 70 kD were measured using double monoclonal antibody assays. RESULTS: Significant concentrations of whole antigen from cat, dog, oak, Alternaria, and Cladosporium were detected. Between 0.1 and 18 microg of Der f1 and Der p1 per gram of dust were also measured. Alt a 1 and Alt a 70 kD levels varied between 3.0 and 1000 U/g of dust. Significant positive correlations were observed in levels of dust mite and Alternaria allergen for patients with an evaluation of asthma. CONCLUSIONS: We found measurable levels of fungal antigens (Alternaria, Cladosporium), mite antigens, and animal antigens (dog and cat) in the majority of dust samples in this self-selected set of allergy clinic patients. Specific allergens Alt a 1, Alt a 70kD, and Der p 1 were significantly higher in the homes of asthmatic patients when compared with patients seen for reasons other than asthma. These studies support the hypothesis that fungal allergen exposure is an important component in the pathogenesis of the clinical condition known as asthma. PMID- 11379803 TI - Asthma severity, atopic status, allergen exposure and quality of life in elderly persons. AB - BACKGROUND: Although asthma can be associated with significant airflow obstruction in those over the age of 65, it is often underdiagnosed and undertreated. OBJECTIVE: To describe severity of asthma, allergy skin test sensitivities, indoor allergen exposures, and the impact on quality of life (QOL) and health status in elderly persons with asthma. METHODS: A cross-sectional data analysis with 80 elderly persons with asthma recruited from medical, geriatric, and allergy/immunology tertiary care centers. Asthma severity was determined by symptoms and measurements of lung function. House dust specimens were collected from mattresses and bedroom carpets and analyzed separately for the major allergens of house dust, using monoclonal antibody-based immunoenzymetric assays. QOL was measured using Juniper's Asthma Quality of Life Questionnaire. Health status was measured using the Short Form Health Survey Medical Outcome Questionnaire which included Ferrans and Powers' Quality of Life Index subscales. RESULTS: Two-thirds of participants had either moderate or severe persistent asthma. Skin tests to a battery of common airborne allergens were positive to at least one allergen in 56 of the 75 participants tested (74.7%). Reservoir dust allergen levels were often high enough to place participants at risk of symptoms or at risk of developing sensitization. Increased asthma severity was associated with significantly lower QOL and a trend toward decreased health status. CONCLUSIONS: Asthma is a significant chronic problem in the elderly. Atopy was common. Asthma severity impacts on these participants' QOL and health status. Results support interventions aimed at identifying allergens precipitating attacks and reducing them in the home. PMID- 11379804 TI - Effect of intranasal challenge with interleukin-6 on upper airway symptomatology and physiology in allergic and nonallergic patients. AB - BACKGROUND: Interleukin-6 (IL-6) is a potent regulator of airway inflammation and an important component of biologic homeostasis. Previously, a temporal relationship between the local elaboration of IL-6 and the development of upper airway symptoms and pathophysiologic findings was reported for patients experimentally infected with influenza A virus or rhinovirus. OBJECTIVE: The objective of this study was to determine the provocative effects of direct, intranasal administration of IL-6 on those symptoms, signs, and pathophysiologic findings that accompany viral upper respiratory infection. METHODS: In this double-blind, placebo-controlled, crossover trial, 10 symptomatic allergic, 10 asymptomatic allergic, and 10 nonallergic adult patients were pretreated with intranasal histamine and, after 15 minutes, were challenged with repeated doses of placebo (saline) or with increasing doses (0, 0.01, 0.1, and 1 microg/mL) of recombinant IL-6 at 20-minute intervals, during randomized paired sessions. Symptom scores, sneeze and cough counts, nasal secretion weights, nasal conductance (rhinomanometry), middle ear pressure (tympanometry), Eustachian tube function (sonotubometry), and pulmonary function (spirometry) were evaluated before and after the histamine challenge, after each dose of IL-6 or placebo, and then at 90 minutes and 2, 3, 4, 6, and 24 hours. RESULTS: At the doses used, intranasal challenge with IL-6 was well tolerated. At the 90-minute postchallenge endpoint, a significant effect of challenge substance and group assignment was documented for nasal secretion weight. Paired comparisons showed that the effect was greater for the allergic patients when compared with the nonallergic patients. There were no differences between placebo and IL-6 challenge for any of the other measured parameters. CONCLUSIONS: These results show that local IL-6 at relatively low doses can provoke increased nasal secretions in patients with allergic rhinitis. PMID- 11379805 TI - Comparison of the cost-effectiveness of budesonide and sodium cromoglycate in the management of childhood asthma in everyday clinical practice. AB - BACKGROUND: Budesonide and sodium cromoglycate are both recommended as maintenance therapy for childhood asthma. OBJECTIVE: To compare the cost effectiveness of these two treatment strategies in clinical practice, in an open label, pharmacoeconomic clinical trial. METHODS: Health economics were evaluated in 138 children, ages 5 to 11 years, with unstable asthma not previously treated with corticosteroids or cromones. The asthma was stabilized during 4 to 6 weeks with budesonide 200 to 400 microg twice daily. The children were then randomly allocated to one of the two treatment strategies aiming at maintaining asthma control for 12 months; budesonide 400 microg/day (N = 69) or sodium cromoglycate 60 mg/day (N = 69). If asthma control was judged unsatisfactory, the doses were increased or the children were switched to the alternate treatment. RESULTS: In children continuing on the same treatment, the degree of asthma control was similar in the two groups at study end. To maintain asthma control, 42% of cromoglycate children switched to budesonide, and then experienced a 14% increase in symptom-free days. No budesonide patient had to switch therapy because of lack of asthma control. Although not statistically significant, total annual cost per patient was 24% (Swedish kronor 4195; US $487; Euro 485) lower in the budesonide than the cromoglycate group, mainly due to a lower cost for asthma medication. CONCLUSIONS: A budesonide strategy for continued maintenance treatment, after an initial period of stabilizing treatment with budesonide, resulted in lower costs and less drug switches than did a strategy with sodium cromoglycate. PMID- 11379807 TI - Comparison of allergenic components between German cockroach whole body and fecal extracts. AB - BACKGROUND: Cockroaches have been demonstrated to be an etiologic factor in allergic diseases. Further, sensitivity to cockroach places patients with asthma at risk for exacerbations that require emergency medical care. OBJECTIVE: This study compared the differences in allergenic components between German cockroach whole body and German cockroach fecal extracts (GWBE and GFE). METHODS: Patients with asthma and/or allergic rhinitis were skin prick tested with German cockroach extract (Bayer Corporation, West Haven, CT). Serum specimens from these patients, 25 with positive skin tests and 8 with negative tests, were used for the ELISA and immunoblot experiments. RESULTS: By ELISA, 72% (18 of 25) and 60% (15 of 25) of positive responders' sera showed IgE antibodies to GWBE and GFE, respectively, and the IgE levels to GWBE were highly correlated with those to GFE (r = .84, P < .01). In inhibition ELISA experiments, extensive cross-reactivity was observed between GWBE and GFE, slight cross-reactivity between GWBE and Dermatophagoides farinae, and no cross-reactivity between GFE and D. farinae. The two-site monoclonal antibody ELISA detected more of the German cockroach major allergens in GFE compared with GWBE; 6.2 times (2420 vs 390 U/mL) for Bla g 1 and 3 times (15.32 vs 5.07 microg/mL) for Bla g 2. In the immunoblot comparison of patients' sera, the IgE antibodies binding to GWBE were apparently different from those binding to GFE in all the positive responders' sera; eg, 50% or more of the 25 positive responders' sera reacted to 43- to 67-kDa proteins in GWBE and to 28- to 30-kDa proteins in GFE, respectively. No IgE antibodies bound to components in GWBE and GFE in the 8 negative responders' sera. CONCLUSIONS: There are major differences between the allergenic components of GWBE and GFE. Based on the amounts of major allergens (Bla g 1, Bla g 2), German cockroach feces are a more important source of allergen than the whole body in respiratory allergic diseases. PMID- 11379806 TI - A sensitive reverse ELISA for the measurement of specific IgE to Der p 2, a major Dermatophagoides pteronyssinus allergen. AB - BACKGROUND: Epidemiologic studies have shown that the presence of IgE antibodies to house dust mite and other indoor allergens is an important risk factor for asthma. OBJECTIVE: The aim of this study was to develop a reverse ELISA (rELISA) for measuring specific IgE to Der p 2, a major Dermatophagoides pteronyssinus (Dpt) allergen, as a potential tool for followup of allergen immunotherapy. METHODS: Recombinant Der p 2 allergen or a monoclonal antibody to Der p 2 was used to coat plates in conventional ELISA (cELISA) and rELISA, respectively. Sera from 48 asthmatic patients with positive skin prick test (SPT+) to D. pteronyssinus extract were analyzed for total IgE and specific IgE to Der p 2, and the results were compared with a group of 41 SPT asthmatic and 30 SPT- control subjects. RESULTS: The sensitivity of the two assays for Der p 2-specific IgE was 3.9 EU/mL and their specificities were confirmed by inhibition tests, in a dose-dependent manner. There was a significant positive correlation between cELISA and rELISA (r = 0.74; P < 0.0001). However, rELISA was more sensitive than was cELISA, regarding both the positive sera percentage (70.8% vs 52.1%) and the Der p 2-specific IgE levels (28.4 vs 4.5 EU/mL) in SPT+ asthmatic patients. CONCLUSIONS: rELISA has shown to be a sensitive and alternative method for measuring Der p 2-specific IgE without using radioactive techniques. Detection of specific IgE to major allergens and relevant peptides, and identification of B cell epitopes in allergens will provide valuable information for the design of allergen analogs and peptides for immunotherapy. PMID- 11379812 TI - High IgD could be a nonpathogenetic diagnostic marker of the hyper-IgD and periodic fever syndrome. PMID- 11379809 TI - Characterization of aerosol output from various nebulizer/compressor combinations. AB - OBJECTIVES: Different commercially available nebulizers and compressors are available. However, the optimal combination for drug delivery is unknown. METHODS: Flow rates of five different compressors (n = 3/compressor) tested alone and in combination with five different commercial nebulizers (n = 9 of each brand of nebulizer) were evaluated. Thereafter, the performances of the different nebulizers were evaluated using 2.5 mg albuterol solution (0.5 mL) added to 2.5 mL saline at flow rates of 2, 3, 4, and 5 L/minute using a laser particle analyzer. Volume median diameter and percentage of particles in the respirable range (1-5 microm) were calculated from this data. Time for nebulization (in seconds) and residual volume (in milliliters) were also recorded. RESULTS: The mean flow rates for the compressors evaluated without a nebulizer attached ranged from 6.6 L/minute (LifeCare Freedom-neb; LifeCare International, Lafayette, CO) to 12.2 L/minute (DeVilbiss Pulmo-Aide; DeVilbiss Health Care, Somerset, PA). Flow rates for the nebulizer/compressor combinations ranged from 2.08 L/minute (Pari LC Jet Proneb; Pari Respiratory Equipment, Richmond, VA) to 5.42 L/minute (Puritan Bennett Raindrop; Puritan Bennett, Lenexa, KS/Omron Compare; Omron, Health Care,Vernon Hills, IL). Using the repeated measure ANOVA model, the interaction between flow rate and device was significant (P < 0.001) for both percentage of particles in the respirable range and log volume median diameter. It was observed that the percentage of particles in the respirable range for the Pari LC Jet did not increase across flow rates in contrast to the other 4 nebulizers. All comparisons to the Pari LC Jet at 2 L/minute were significant. CONCLUSIONS: Marked variability exists in the flow rates among different commercially available compressors used for home nebulization of inhaled pulmonary medications. Different nebulizer/compressor combinations have markedly different performance characteristics which could result in different efficacy and safety profiles of the medications being administered via these devices. We recommend that this type of information be used as a starting point for selecting different nebulizer/compressor combinations. Further clinical evaluation is warranted. PMID- 11379808 TI - Long-term safety and efficacy of a chlorofluorocarbon-free beclomethasone dipropionate extrafine aerosol. AB - BACKGROUND: Beclomethasone dipropionate (BDP) extrafine aerosol, a newly developed pressurized metered dose inhaler (pMDI) with a hydrofluoroalkane-134a (HFA) propellant (HFA-BDP; Qvar, 3M Pharmaceuticals, St. Paul, MN), has been shown to be effective in controlling asthma symptoms at approximately half the daily dose of chlorofluorocarbon (CFC)-BDP. OBJECTIVE: This study evaluated the long-term efficacy and safety of switching patients with asthma maintained on a stable dose of CFC-BDP pMDI to therapy with HFA-BDP pMDI at approximately half their previous daily dose of CFC-BDP. METHODS: This was an open-label, randomized, parallel-group multicenter trial. Patients with at least a 6-month history of asthma whose symptoms were controlled on CFC-BDP, 400 to 1600 microg daily, during a 2-week run-in period were randomized in a 1:3 ratio to CFC-BDP at the same daily dose or HFA-BDP at approximately half the daily dose of CFC-BDP for 12 months. RESULTS: A total of 473 patients were randomized: 354 to HFA-BDP, 119 to CFC-BDP. There were no statistically significant differences between groups in mean change from baseline in morning (AM) peak expiratory flow rate or forced expiratory volume in one second throughout the study. There were no consistent differences between treatment groups in individual asthma symptoms or daily beta2-agonist use during the study. There was an increase in the percentage of symptom-free days between baseline and month 12 in the HFA-BDP group (11.5%) and the CFC-BDP group (4.6%). No statistically significant differences in serum osteocalcin levels or adverse events were seen during the study or in AM plasma cortisol levels at month 12. CONCLUSIONS: Asthma control was maintained in patients switched from a stable dose of CFC-BDP (400 to 1600 microg daily) to HFA BDP at approximately half the CFC-BDP dose (200 to 800 microg daily), and was maintained over the next 12 months. HFA-BDP demonstrated a similar safety profile to CFC-BDP; there were no differences between the agents with regard to systemic effects. PMID- 11379811 TI - Severe food allergies by skin contact. AB - BACKGROUND: Ingestion is the principal route for food allergens, yet some highly sensitive patients may develop severe symptoms upon skin contact. CASE REPORT: We describe five cases of severe food allergic reactions through skin contact, including inhalation in one. METHODS: The cases were referred to a university allergy clinic, and evaluation comprised detailed medical history, physical examination, skin testing, serum total and specific IgE, and selected challenges. RESULTS: These cases were found to have a strong family history of allergy, early age of onset, very high total serum IgE level, and strong reactivity to foods by skin prick testing or RAST. Interestingly, reactions occurred while all five children were being breast-fed (exclusively in four and mixed in one). CONCLUSIONS: Severe food allergic reactions can occur from exposure to minute quantities of allergen by skin contact or inhalation. Food allergy by a noningestant route should be considered in patients with the above characteristics. PMID- 11379810 TI - A randomized, double-blind comparison of beclomethasone dipropionate extrafine aerosol and fluticasone propionate. AB - BACKGROUND: Inhaled corticosteroids provide first-line treatment for asthma. An advance to improve potency was to produce new molecules with increased glucocorticoid receptor affinity (eg, fluticasone propionate [FP]). An alternative is to deliver more medication to both the large and small airway inflammation of asthma by using an extrafine aerosol (eg, beclomethasone dipropionate extrafine aerosol [BDP-extrafinel). OBJECTIVE: To demonstrate clinical equivalence of BDP-extrafine (400 microg daily) and FP (400 microg daily) in symptomatic asthmatic patients over the course of 6 weeks. METHODS: This was a double-blind, double-dummy, parallel-group, multicenter, 6-week study in adults with asthma taking conventional FP 100 to 250 microg daily or equivalent, and displaying signs/symptoms of active disease requiring additional therapy. RESULTS: Eighty-eight patients were randomized to BDP-extrafine (and FP placebo) and 84 to FP (and BDP-placebo). There were no significant differences between treatments with respect to symptom control, as evidenced by mean change from baseline in percentage days without asthma symptoms/nights without sleep disturbance observed at weeks 1 to 2, 3 to 4, or 5 to 6. Mean changes from baseline in AM PEFR at weeks 5 to 6 for BDP-extrafine (19.0) and FP (30.5) were equivalent (P = 0.022 for equivalence). There were significant (P < 0.001) within treatment-group differences in mean change from baseline in AM PEFR at weeks 1 to 2 for both treatments. There was no difference in the incidence of patients reporting at least one adverse event during the study (BDP-extrafine 41%; FP 37%). Mean percentage change from baseline for AM plasma cortisol at week 6 was + 17.7% for BDP-extrafine and +4.2% for FP (P = 0.066 for difference). CONCLUSIONS: BDP-extrafine and FP at doses of 400 microg daily provided equivalent asthma control in patients with symptomatic asthma and exhibited similar safety profiles. PMID- 11379813 TI - Chronic rhinosinusitis. PMID- 11379814 TI - Kentucky bluegrass. PMID- 11379815 TI - Cortical synapse loss in progressive supranuclear palsy. AB - Cortical synapse loss, the probable substrate of cognitive impairment in Alzheimer disease (AD), has not previously been evaluated in progressive supranuclear palsy (PSP). Hypothesizing that synapse loss would be greater in demented than non-demented PSP patients, we examined synaptophysin concentrations in 8 cases of PSP (5 demented and 3 nondemented cases). We found a decrease in mean synaptophysin concentration in these 8 cases in frontal, temporal, and parietal lobes, and in cerebellum, compared to the means in corresponding lobes of 16 controls. The decreases were similar to those in 28 cases of AD, but not as great. We determined synaptophysin concentration from motor cortex in only 4 of our PSP cases, 2 demented and 2 non-demented. The average concentrations in these 4 cases were lower than in AD motor cortex; both were lower than controls. When demented and non-demented PSP cases were compared, neocortical synaptophysin concentrations in non-demented PSP cases were lower than in demented cases. There appears to be a link between AD and PSP, in that synapse loss is found in both. However, the basis and significance of the prominent neocortical synapse loss in PSP, especially in non-demented subjects, remain to be explored. PMID- 11379816 TI - Delayed peripheral nerve degeneration, regeneration, and pain in mice lacking inducible nitric oxide synthase. AB - Inducible nitric oxide synthase (iNOS) may be a critical factor in the repair of injured tissues. In mice lacking iNOS we observed abnormalities in how the peripheral nerve responds to each of 3 fundamental types of injury: chronic constriction partial nerve injury (a model of neuropathic pain), nerve crush, and nerve transection. In each type of injury, mice lacking iNOS had evidence of a regenerative delay, preceded by slowing of myelinated fiber Wallerian degeneration (WD). In wild-type mice, iNOS immunoreactivity and the presence and upregulation of its mRNA were demonstrated distal to injury, but neither was observed in the knockout mice. Slowed WD was suggested by the abnormal persistence of apparent myelinated fiber profiles distal to the injury zones in mice lacking iNOS compared to wild-type controls. In mice lacking iNOS there were fewer regenerating myelinated fibers, smaller caliber regenerating fibers, and slowed reinnervation of muscle endplates distal to the injury zone. Slowed degeneration was also associated with normal initiation but delayed expression of neuropathic pain. Our findings highlight important relationships among nitric oxide, WD, neuropathic pain, and axon regeneration. PMID- 11379817 TI - Apoptotic and anti-apoptotic mechanisms following spinal cord injury. AB - A number of studies have provided evidence that cell death from moderate traumatic spinal cord injury (SCI) is regulated, in part, by apoptosis that involves the caspase family of cysteine proteases. However, little or no information is available about anti-apoptotic mechanisms mediated by the inhibitors of apoptosis (IAP) family of proteins that inhibit cell death pathways. In the present study, we examined caspase and IAP expression in spinal cords of rats subjected to moderate traumatic injury. Within 6 h after injury, caspase-8 and-9 (2 initiators of apoptosis) were predominantly present in gray matter neurons within the lesion epicenter. By 3 days following spinal cord injury (SCI), caspase-8 and-9 immunoreactivity was localized to gray and white matter cells, and by 7 days following SCI, both upstream caspases were expressed in cells within white matter or within foamy macrophages in gray matter. Caspase 3, an effector caspase, was evident in a few fragmented cells in gray matter at 24 h following injury and then localized to white matter in later stages. Thus, distinct patterns of caspase expression can be found in the spinal cord following injury. XIAP, cIAP-1, and cIAP-2, members of the IAP family, were constitutively expressed in the cord. Immunoblots of spinal cord extracts revealed that the processed forms of caspases-8 and-9 and cleavage of PARP are present as early as 6 h following trauma. The expression of caspases corresponded with the detection of cleavage of XIAP into 2 fragments following injury. cIAP-1 and cIAP-2 expression remained constant during early periods following SCI but demonstrated alterations by 7 days following SCI. Our data are consistent with the idea that XIAP may have a protective role within the spinal cord, and that alteration in cleavage of XIAP may regulate cell death following SCI. PMID- 11379818 TI - Elevation of LDL receptor-related protein levels via ligand interactions in Alzheimer disease and in vitro. AB - The low-density lipoprotein (LDL) receptor-related protein (LRP) is a multifunctional receptor in the CNS that binds both apolipoprotein E (apoE) and activated alpha2-macroglobulin (alpha2M*); all 3 proteins are genetically associated with Alzheimer disease (AD). In this study we found an 85% increase in LRP levels in human AD brain frontal cortex, along with an increased level of the LRP ligands, apoE, and alpha2M. We speculated that LRP levels might be increased in response to the increased levels of its ligands, apoE, and alpha2M*. To test this hypothesis we examined the effects of alpha2M* on LRP in primary cultures. Treatment of neurons with alpha2M* significantly increased LRP levels (by 92%). This increase was prevented by coculture with receptor-associated protein (RAP), which blocks binding of LRP ligands to LRP Native alpha2M or RAP alone did not change LRP levels in vitro. We also found that alpha2M* stimulated activation of astrocytes in vitro and promoted the levels of LRP by 65%. These data indicate 1) the LRP ligand alpha2M* increases levels of LRP in primary neuronal and astrocytic cultures, 2) alpha2M*-induction of LRP levels in vitro depends on binding to LRP, and 3) LRP levels are increased in AD brain, perhaps in response to the increased levels of alpha2M. PMID- 11379819 TI - O-Glycosylation in sprouting neurons in Alzheimer disease, indicating reactive plasticity. AB - Reactive plasticity, including axonal and dendritic sprouting and reactive synaptogenesis, has been proposed to contribute to the pathogenesis of several neurological disorders. This work was aimed at identifying the possible role of protein glycosylation in the brain from patients with Alzheimer disease (AD), using lectin histochemistry, as determinants of reactive plasticity. Results indicate an increase in the production of cryptic O-glycosidically linked proteins (NeuAcalpha2,6 Galbeta1,3GalNAcalpha1,0 Ser/Thr or sialyl-T-antigen) in neuritic sprouting in AD brains as determined by positive labeling with Amaranthus leucocarpus (ALL, T-antigen-specific) and Macrobrachium rosenbergii (MRL, specific for NeuAc5,9Ac2) lectins. Immunohistochemistry indicated that lectin staining was specific for the synaptic sprouting process (meganeurites) in AD. These results were confirmed using anti-synaptophysin and anti-GAP 43 antibodies, which recognized meganeurites and dystrophic neurites around amyloid beta deposits. In normal control brains, labeling with the aforementioned lectins was restricted to microvessels. Control experiments with neuraminidase-treated brain samples revealed positivity to the lectin from Arachis hypogaea (PNA), which is specific for galactose. Our results suggest specific O-glycosylation patterns of proteins closely related to neuronal plasticity in AD. PMID- 11379820 TI - Fibroblasts can express glial fibrillary acidic protein (GFAP) in vivo. AB - Neuropathologists use anti-glial fibrillary acidic protein (GFAP) antibodies as specific markers for glial cells, and neurobiologists use GFAP for targeting transgenes to glial cells. Since GFAP has also been detected in non-glial cells, we systematically analyzed GFAP expression in human and murine non-CNS tissues using a panel of anti-GFAP antibodies. In human tissues we confirm previously observed GFAP expression in Schwann cells, myoepithelial cells, and chondrocytes, and show for the first time GFAP expression in fibroblasts of epiglottic and auricular perichondrium, ligamentum flavum, and cardiac valves. In mice we show GFAP expression in Schwann cells, bone marrow stromal cells, chondrocytes, and in fibroblasts of dura mater, skull and spinal perichondrium, and periosteum, connective stroma of oral cavity, dental pulp, and cardiac valves. Anti-GFAP immunoblotting of human non-CNS tissues reveals protein bands with a molecular mass ranging between approximately 35 and approximately 42 kDa. In GFAP-v-src transgenic mice, whose oncogenic v-src transgene transforms GFAP expressing cells, non-CNS tumors originate from fibroblasts. We conclude that human and murine fibroblasts can express GFAP in vivo. The somatic distribution of GFAP expressing fibroblasts indicates origin from the neural crest. Development of non CNS tumors from fibroblasts in GFAP-v-src mice functionally confirms GFAP expression in these cells. PMID- 11379821 TI - Apoptosis, neuronal maturation, and neurotrophin expression within medulloblastoma nodules. AB - Nodular/desmoplastic medulloblastomas are a well-established histopathological subtype containing reticulin-free nodules or "pale islands' that are comprised of cells with round "neurocytic" nuclei and abundant cytoplasm. Significant neuronal maturation occurs within nodules. We used immunohistochemistry to evaluate neuronal differentiation in the nodules of 6 of these tumors. The neuronal markers NeuN, synaptophysin, and MAP-2 were identified in the "pale islands" of all 6 nodular medulloblastomas examined, and high and medium molecular weight nonphosphorylated neurofilaments were detected in 2 of the 6 cases. We also observed collections of apoptotic cells within nodules. Given the known role of neurotrophin signaling in neuronal maturation and apoptosis, we analyzed immunohistochemically the distribution of neurotrophin receptors TrkA and TrkC and their primary ligands NGF and NT3 in 14 nodular medulloblastomas. TrkA and TrkC were detected in 13 and 10 cases, respectively, and were predominantly localized within nodules. NGF and NT3 were distributed diffusely with some nodular accentuation. The localized expression of Trk receptors within nodules of desmoplastic medulloblastomas suggests neurotrophin signaling is involved in the apoptosis and neuronal differentiation in medulloblastomas. We also examined expression of p53 and BCL-2 in these tumors; both were prominent in internodular regions but only weakly expressed within nodules. Trk receptors, p53, and BCL-2 are all expressed during development of the normal cerebellum. Interestingly, the immunohistochemical expression profile of these proteins in the differentiating nodules of medulloblastomas is in many ways similar to their expression in the developing cerebellum. Thus similar signaling pathways may be operational in cerebellar development and medulloblastoma tumor differentiation. PMID- 11379822 TI - Acute hypoxia-induced alterations of calbindin-D28k immunoreactivity in cerebellar Purkinje cells of the guinea pig fetus at term. AB - Purkinje cells (PCs) are vulnerable to hypoxic/ischemic insults and rich in calcium and calcium-buffering/sequestering systems, including calcium-binding proteins (CaBPs). Calbindin-D28k is an EF-hand CaBP, which is highly expressed in PCs where it acts primarily as a cellular Ca++ buffer. Elevation of [Ca++] in the cytosol and nuclei of PCs is pivotal in hypoxic/ischemic cell death. We hypothesize that hypoxia results in decreased concentration, or availability of calbindin-D28k in PCs, thereby decreasing their buffering capacity and resulting in increase of intracellular and intranuclear [Ca++]. Cerebellar tissues from normoxic fetuses were compared to fetuses obtained from term pregnant guinea pigs exposed to hypoxia [7% FiO2] for 60 min. The pregnant guinea pigs were either killed upon delivery immediately following hypoxia (Hx0h) or were subsequently allowed to recover for 24 h (Hx24h) or 72 h (Hx72h). Fetal brain hypoxia was documented biochemically by a decrease in brain tissue levels of ATP and phosphocreatine. Compared to normoxic fetuses, there is a predominantly somatodendritic loss or decrease of calbindin-D28k immunohistochemical staining in PCs of Hx0h (p < 0.005), Hx24h (p < 0.05), and Hx72h (p < 0.005) fetuses. Hypoxia-induced alterations of calbindin-D28k immunoreactivity are qualitatively similar at all time points and include a distinctive intranuclear localization in subpopulations of PCs. A similar trend is demonstrated by immunoblotting. Subpopulations of TUNEL+/calbindin-D28k- PCs lacking morphologic features of apoptosis or necrosis are demonstrated in Hx24h and Hx72h fetuses. The present study demonstrates an abrogating effect of perinatal hypoxia on calbindin-D28k immunoreactivity in cerebellar PCs. The perturbation of this Ca++ buffer protein in hypoxia-induced neuronal injury may herald delayed cell death or degeneration. PMID- 11379823 TI - Variant Alzheimer disease with spastic paraparesis: neuropathological phenotype. AB - Variant Alzheimer disease (varAD) is clinically characterized by the combination of presenile dementia with spastic paraparesis and is caused by certain mutations of the presenilin 1 (PS-1) gene. We now present the unusual neuropathological phenotype of varAD as seen in 5 affected members of the original Finnish family with a genomic deletion encompassing exon 9 of the PS-1 gene. Their primary and association cortices and hippocampus showed a profusion of eosinophilic, roundish structures with distinct borders termed "cotton wool" plaques (CWPs). The CWPs were immunoreactive for Abeta42/43 but weakly or not at all for Abeta40 isoforms of the amyloid beta peptide (Abeta). They were devoid of a congophilic core, and fibrillar amyloid could not be identified within them by electron microscopy. Confocal microscopy showed reduced density of axons within individual CWPs and only few CWP-related PHF-tau-positive dystrophic neurites. CWPs were particularly numerous in the medial motor cortex representing the lower extremities, and degeneration of the lateral corticospinal tracts was observed at the level of the medulla oblongata and the spinal cord. In addition to the predominant CWPs, variable numbers of diffuse and cored plaques were found in the cerebral cortex. Diffuse and non-neuritic cored amyloid plaques but no CWPs occurred in the cerebellum. In conclusion, varAD in this Finnish family is distinct from classic AD because of the degeneration of lateral corticospinal tracts, predominance of CWPs devoid of fibrillar amyloid cores in the cerebral cortex, and presence of non-neuritic amyloid plaques in the cerebellum. PMID- 11379824 TI - The dorsal root ganglia in adrenomyeloneuropathy: neuronal atrophy and abnormal mitochondria. AB - Adrenomyeloneuropathy (AMN), a disease of spinal cord, brain, adrenal, and testis, mostly affects men with spastic paraparesis or ataxia beginning in their second or third decade. The spinal cord displays bilateral, usually symmetrical, long tract degeneration particularly of the gracile tract in a "dying-back" pattern. The available data strongly indicate that the fundamental lesion in AMN is an axonopathy or neuronopathy. We compared lumbar dorsal root ganglia (DRG) from 3 AMN patients to 6 age-matched controls histologically, morphometrically, immunohistochemically, and ultrastructurally. There was no apparent neuronal loss, necrosis or apoptosis, nor obvious atrophy; nodules of Nageotte were sparse in both groups. The morphometric studies, however, did reveal neuronal atrophy with a decrease in the number of large neurons and a corresponding increase in neurons less than 2,000 microm2, especially in the 1,500-1,999 microm2 range. No consistent immunohistochemical differences were observed, and no specific cell type appeared to be lost. Many mitochondria in the AMN neurons demonstrated lipidic inclusions; this raises the possibility that, in addition to the well known peroxisomal defect, impaired mitochondrial function may lead to a failure of ATP-dependent axoplasmic transport in AMN spinal tracts with consequent "dying back" axonal degeneration. The observation that the DRG parent neurons of the degenerate gracile tracts in AMN undergo atrophy and do not display appreciable evidence of cell death, even at autopsy, provides a wide window of opportunity for the development of therapeutic strategies to combat or prevent this myeloneuropathy. PMID- 11379825 TI - Evaluating characteristics of patient selection and dropout rates. AB - Patient selection and dropout rates can affect the results of a clinical trial. Long lists of exclusions in the selection of patients for clinical trials reduce the possibility of examining treatment responses for heterogeneity and make recruitment difficult. In many cases, a pool of 100 potential subjects may yield only 2 or 3 qualified participants, a fact that raises the issue of generalizability of results. Dropouts should be carefully defined in advance and can be used as dependent variables for the comparison of different treatments. This article discusses some of the sampling characteristics (gender, age, diagnosis, inpatient/ outpatient status, prior neuroleptic use, and symptom severity) and dropout rates in 5 recent comparative clinical trials of atypical antipsychotics. PMID- 11379826 TI - Clinical trial design issues in schizophrenic research. AB - Research studies are an indispensable tool in the Food and Drug Administration approval process. Becoming familiar with different research study designs helps one to understand and interpret the results of clinical trials. Research studies are undertaken with 2 major goals in mind: (1) to get a drug into registration (regulatory needs) and (2) to understand the clinical application of medicines (clinical application needs). This article discusses the 2 major goals of research studies, the design issues associated with those goals, and the design features of several recent comparative studies of atypical antipsychotics. PMID- 11379827 TI - Evaluating clinical trial data: outcome measures. AB - Schizophrenia is too often a severely disabling disease, and the discovery of interventions that can ease or eliminate symptoms without troubling side effects has long been the goal of schizophrenia research. In this endeavor, researchers, clinicians, and patients all desire an optimal outcome; outcome measures, which measure the relative success or failure of an intervention, are accordingly important. In addition, the costs of pharmacologic interventions-particularly of the atypical antipsychotics--in schizophrenia make the optimal measurement of treatment outcomes critical. Sometimes outcomes are focused on minimizing costly events, such as rehospitalization, rather than focusing on patient-oriented outcomes. This article discusses the outcome measures employed in 5 clinical trials comparing atypical antipsychotics, examining their usefulness and suggesting types of outcome measures that may be useful in the future. PMID- 11379828 TI - Dose selection and comparator drugs in schizophrenia research. AB - Dose selection and comparator drugs are important methodological issues in the evaluation of clinical data. Investigators rely heavily on dose selection in schizophrenia research, particularly when making comparisons between and across groups of compounds. In reality, however, the criteria for selecting a dose of drug in research trials are rather crude. Dosing and comparator issues vary enormously, depending on several factors. This article will discuss some of the dose-selection issues in schizophrenia research and the doses of antipsychotic drugs used in 5 recent comparative clinical trials. PMID- 11379829 TI - Introduction: evaluating clinical trial data from schizophrenia research. PMID- 11379830 TI - The statistical comparison of clinical trials. AB - Being able to compare clinical trials using statistically proven analyses is essential, but certain protocols must be followed if such a comparison is to be made. The difficulty of making meaningful statistical comparisons is illustrated in 5 clinical trials comparing atypical antipsychotics. Certain judgments can be made on the basis of internal and external validity, for example, but there are many other areas--effect size, for example--in which it is impossible to make any statistical comparisons across these trials because they were not conducted in a uniform fashion. In the final analysis, it is doubtful that the differences between atypical antipsychotics covered in these trials are greater than those due to chance. PMID- 11379831 TI - Interpretations and conclusions in the clinical trial. AB - Interpreting the results of a clinical trial and reaching conclusions are matters of assessing the integrity of the trial's design and methods and the validity of the findings and drawing inferences to make conclusions that will guide health care decisions; these decisions are the ultimate goal of the clinical trial. This article examines 5 clinical trials of atypical antipsychotic agents with particular attention to aspects of their validity and refers to other articles in this supplement when appropriate. Finally, the Behavioral Health Care Enterprise Model, designed to help systematize the process of drawing inferences and implications from these and other clinical trials and assessing their impact on various sectors of the health care system, is introduced. PMID- 11379832 TI - Hypothesis and hypothesis testing in the clinical trial. AB - The hypothesis provides the justification for the clinical trial. It is antecedent to the trial and establishes the trial's direction. Hypothesis testing is the most widely employed method of determining whether the outcome of clinical trials is positive or negative. Too often, however, neither the hypothesis nor the statistical information necessary to evaluate outcomes, such as p values and alpha levels, is stated explicitly in reports of clinical trials. This article examines 5 recent studies comparing atypical antipsychotics with special attention to how they approach the hypothesis and hypothesis testing. Alternative approaches are also discussed. PMID- 11379833 TI - The need for clinically relevant research on treatment-resistant depression. PMID- 11379834 TI - Why drugs and hormones may interact in psychiatric disorders. PMID- 11379835 TI - Bupropion sustained release for bereavement: results of an open trial. AB - OBJECTIVE: The present study was conducted to assess whether DSM-IV-defined bereavement responds to bupropion sustained release (SR). METHOD: Twenty-two subjects who had lost their spouses within the previous 6 to 8 weeks and who met DSM-IV symptomatic/functional criteria for a major depressive episode were evaluated. Subjects completed the Hamilton Rating Scale for Depression (HAM-D), the Clinical Global Impressions scale, the Texas Revised Inventory of Grief, and the Inventory of Complicated Grief at baseline and follow-up. Subjects were treated with bupropion SR, 150 to 300 mg/day, for 8 weeks. RESULTS: Improvement was noted in both depression and grief intensity. For the intent-to-treat group. 59% experienced a reduction of > or = 50% on HAM-D scores. The correlations between changes in the HAM-D scores and the grief scale scores were high, ranging from 0.61 (p = .006) to 0.44 (p = .054). CONCLUSION: Major depressive symptoms occurring shortly after the loss of a loved one (i.e., bereavement) appear to respond to bupropion SR. Treatment of these symptoms does not intensify grief; rather, improvement in depression is associated with decreases in grief intensity. The results of this study challenge prevailing clinical wisdom that DSM-IV-defined bereavement should not be treated. Larger, placebo-controlled studies are indicated. PMID- 11379836 TI - Factors influencing acute weight change in patients with schizophrenia treated with olanzapine, haloperidol, or risperidone. AB - OBJECTIVE: Clinical factors predicting weight change in patients with schizophrenia and related disorders during acute treatment with the antipsychotic drugs olanzapine, risperidone, and haloperidol were sought through retrospective analyses. METHOD: Six-week body-weight data from 2 trials, study 1 comparing olanzapine and haloperidol (N = 1,369) and study 2 olanzapine and risperidone (N = 268), were analyzed. Effects of 8 clinically relevant covariates--therapy, clinical outcome (Brief Psychiatric Rating Scale), baseline body mass index (BBMI), increased appetite, age, gender, race, and dose--on weight were compared. RESULTS: In study 1, olanzapine (vs. haloperidol) therapy, better clinical outcome, lower BBMI, and nonwhite race significantly affected weight gain. Effects of increased appetite and male gender on weight gain were significant for olanzapine but not for haloperidol. In study 2, better clinical outcome, lower BBMI, and younger age significantly affected weight gain. Increased appetite was more frequent during olanzapine treatment than during haloperidol, but not significantly different from risperidone. Significant differences in effect on weight change were found between olanzapine and haloperidol but not between olanzapine and risperidone. No evidence was found that lower antipsychotic drug doses were associated with lower weight gain. CONCLUSION: This report identifies predictive factors of acute weight change in patients with schizophrenia. Similar factors across antipsychotic drugs in predicting greater weight gain included better clinical outcome, low BBMI, and nonwhite race. Factors differing between conventional (haloperidol) and atypical (olanzapine) agents included increased appetite and gender. Choice of atypical antipsychotic drug (olanzapine vs. risperidone) was of minor importance with regard to influence on acute weight gain. PMID- 11379837 TI - A randomized controlled trial of risperidone in the treatment of aggression in hospitalized adolescents with subaverage cognitive abilities. AB - BACKGROUND: Risperidone is an atypical antipsychotic drug that blocks dopamine as well as serotonin receptor systems. The present study was designed to examine the efficacy and safety of risperidone in a 6-week double-blind, randomized, parallel group design in the treatment of aggression in adolescents with a primary diagnosis of DSM-IV disruptive behavior disorders and with subaverage intelligence. METHOD: We randomly assigned 38 adolescents (33 boys; 10 subjects with slightly subaverage IQ, 14 with borderline IQ, and 14 with mild mental retardation), who were hospitalized for treatment of psychiatric disorders associated with severe aggression, to receive risperidone or placebo. The main efficacy measures were the Clinical Global Impressions-Severity of Illness scale (CGI-S), the modified Overt Aggression Scale (OAS-M), and the Aberrant Behavior Checklist (ABC). Side effects were measured using the Extrapyramidal Symptom Rating Scale (ESRS). RESULTS: The mean daily dose of risperidone at the end of treatment was 2.9 mg (range, 1.5-4 mg). Risperidone, compared with placebo, was associated with significant improvements on the CGI-S (p < .001) and the at school ABC overall and hyperactivity scales (p < .05). During a 2-week washout following the 6-week trial, a statistically significant worsening was found in the risperidone group on the CGI-S scale, the OAS-M. and the ABC. Extrapyramidal symptoms were absent or very mild during risperidone treatment. Transient tiredness was present in 11 (58%) of 19 drug-treated subjects. Other untoward effects included sialorrhea, nausea, and slight weight gain (mean = 3.5% of body weight in the risperidone group). No clinically relevant changes were found in laboratory parameters, electrocardiogram, heart rate, or blood pressure. CONCLUSION: These results suggest that risperidone may be effective for severe aggression in adolescents with disruptive behavior disorders and subaverage intelligence, and these results are consistent with reports suggesting its effectiveness for treating severe aggression in adolescents in general. PMID- 11379838 TI - Antidepressant-induced mania in bipolar patients: identification of risk factors. AB - BACKGROUND: Concerns about possible risks of switching to mania associated with antidepressants continue to interfere with the establishment of an optimal treatment paradigm for bipolar depression. METHOD: The response of 44 patients meeting DSM-IV criteria for bipolar disorder to naturalistic treatment was assessed for at least 6 weeks using the Montgomery-Asberg Depression Rating Scale and the Bech-Rafaelson Mania Rating Scale. Patients who experienced a manic or hypomanic switch were compared with those who did not on several variables including age, sex, diagnosis (DSM-IV bipolar I vs. bipolar II), number of previous manic episodes, type of antidepressant therapy used (electroconvulsive therapy vs. antidepressant drugs and, more particularly, selective serotonin reuptake inhibitors [SSRIs]), use and type of mood stabilizers (lithium vs. anticonvulsants), and temperament of the patient, assessed during a normothymic period using the hyperthymia component of the Semi-structured Affective Temperament Interview. RESULTS: Switches to hypomania or mania occurred in 27% of all patients (N = 12) (and in 24% of the subgroup of patients treated with SSRIs [8/33]); 16% (N = 7) experienced manic episodes, and 11% (N = 5) experienced hypomanic episodes. Sex, age, diagnosis (bipolar I vs. bipolar II), and additional treatment did not affect the risk of switching. The incidence of mood switches seemed not to differ between patients receiving an anticonvulsant and those receiving no mood stabilizer. In contrast, mood switches were less frequent in patients receiving lithium (15%, 4/26) than in patients not treated with lithium (44%, 8/18; p = .04). The number of previous manic episodes did not affect the probability of switching, whereas a high score on the hyperthymia component of the Semistructured Affective Temperament Interview was associated with a greater risk of switching (p = .008). CONCLUSION: The frequency of mood switching associated with acute antidepressant therapy may be reduced by lithium treatment. Particular attention should be paid to patients with a hyperthymic temperament, who have a greater risk of mood switches. PMID- 11379839 TI - Effects of nefazodone on body weight: a pooled analysis of selective serotonin reuptake inhibitor- and imipramine-controlled trials. AB - BACKGROUND: Evidence suggests that the newer antidepressant drugs may differ with respect to their effects on body weight, especially during long-term treatment. However, the published data about treatment-emergent weight change with the newer antidepressants are limited. Most reports of unexpected selective serotonin reuptake inhibitor (SSRI)-associated weight gain are anecdotal or from small controlled trials. To determine if differences exist among the newer antidepressants, the authors retrospectively analyzed data from clinical trials comparing nefazodone with SSRIs and with imipramine. METHOD: Weight change data supplied by Bristol-Myers Squibb from 6 completed clinical trials comparing the antidepressant nefazodone (N = 523) with 3 SSRIs, fluoxetine, sertraline, and paroxetine (N = 513), as well as 3 trials comparing nefazodone (N = 225) with the tricyclic antidepressant imipramine (N = 224) were analyzed. In all studies, nefazodone was found to be equal in efficacy to the comparator antidepressants. Studies that included both acute and long-term treatment phases were included in the analysis. Acute phases of the trials lasted either 6 or 8 weeks, and long term phases varied in duration from 16 to 46 weeks. The analysis included summarizing the number and percentage of patients in each group with a > or = 7% change in body weight from baseline at any point in the long-term and acute phases, at endpoint, and at week 16 of the long-term phases. RESULTS: Using 7% or greater weight change as the measure of clinical significance, 4.3% of SSRI treated patients had lost weight at any point in the acute phase versus 1.7% of those treated with nefazodone (p = .017). However, at any point during the long term phase, significantly more SSRI-treated patients than nefazodone-treated patients showed a significant increase in body weight (17.9% vs. 8.3%; p = .003). At any point in the acute phase, significantly more imipramine-treated patients than nefazodone-treated patients had a 7% or greater increase in body weight (4.9% vs. 0.9%; p = .027), and for the long-term phase the comparison yielded 24.5% versus 9.5%. The difference during the long-term phase was statistically significant in women (p = .017), but not in men (p = .078) due to the small numbers of men in each group. CONCLUSION: SSRIs caused more weight loss during short-term treatment but more weight gain during long-term treatment. These results lend support to the observation that some antidepressants have a greater expected risk of weight gain than others during long-term therapy. PMID- 11379840 TI - Health-related quality-of-life measure enhances acute treatment response prediction in depressed inpatients. AB - BACKGROUND: Many nonbiological variables are reported to predict treatment response for major depression; however, there is little agreement about which variables are most predictive. METHOD: Inpatient subjects (N = 59) diagnosed with current DSM-IV major depressive disorder completed weekly depressive symptom ratings with the Hamilton Rating Scale for Depression (HAM-D-17) and Beck Depression Inventory (BDI), and weekly health-related quality-of-life (HRQL) ratings with the Quality of Well-Being Scale (QWB). Acute responders were identified by a 50% decrease in HAM-D-17 score from baseline within 4 weeks of medication treatment. Predictor variables were initially chosen from a literature review and then tested for their association with acute treatment response. RESULTS: An initial predictive model including age at first depression, admission BDI score, and melancholia predicted acute treatment response with 69% accuracy and was designated as the benchmark model. Adding the admission QWB index score to the benchmark model did not improve the prediction rate; however, adding the admission QWB subscales for physical and social activity to the benchmark model significantly improved acute treatment response prediction to 86% accuracy (p = .001). CONCLUSION: In addition to being designed for use in cost-effectiveness analyses, the QWB subscales appear to be useful HRQL variables for predicting acute inpatient depression treatment response. PMID- 11379841 TI - Smoking habits in bipolar and schizophrenic outpatients in southern Israel. AB - BACKGROUND: Although rates of cigarette smoking have been found to be higher in schizophrenic and depressed patients than in the general population, data regarding rates in bipolar patients are limited. This study further examines the relationship between bipolar disorder and smoking and compares the rate of smoking in bipolar disorder patients with rates in schizophrenic patients and in the general population. METHOD: Seventy bipolar patients and 64 schizophrenic patients (diagnosed using DSM-IV criteria) treated at the largest specialized public bipolar and schizophrenia clinics in southern Israel were interviewed regarding their smoking habits. The interview included a questionnaire relating to personal information, past and present smoking, and drug abuse and the Fagerstrom scale for nicotine dependence. Data from these patients were also compared with data from the general Israeli population. RESULTS: Data indicate that the rate of smoking does not appear to differ between bipolar (43.0%) and schizophrenic (45.0%) patients, whereas the rate for both patient groups is higher than that for the general Israeli population (27.5%). Smoking intensity was not found to be different between the 2 groups of patients. CONCLUSION: Smoking in patients with schizophrenia was suggested to be related to nicotine cholinergic dysfunction, but this suggestion cannot explain the equally high rates of smoking in bipolar patients. Schizophrenia, bipolar disorder, and smoking may all be related to dopamine transmission, and, therefore, dopaminergic interactions may provide a better explanation for the results. PMID- 11379842 TI - Long-term olanzapine therapy in the treatment of bipolar I disorder: an open label continuation phase study. AB - BACKGROUND: Olanzapine has demonstrated efficacy in the treatment of acute mania in 2 double-blind, placebo-controlled trials. We describe the results of the open label extension from one of these trials. METHOD: In a 3-week, double-blind study of patients with DSM-IV bipolar I disorder, olanzapine was superior to placebo for the treatment of acute manic symptoms. Of the 139 patients who entered the double-blind phase of the 3-week study, 113 patients continued into the 49-week open-label extension. Efficacy measurements including the Young Mania Rating Scale (YMRS), the 21-item Hamilton Rating Scale for Depression (HAM-D-21), the Clinical Global Impressions scale-Bipolar Version, and the Positive and Negative Syndrome Scale and safety measurements including the Simpson-Angus scale, the Barnes Akathisia Scale, and the Abnormal Involuntary Movement Scale were completed throughout. The analysis considered all treatment results, starting with the first olanzapine dose. Adjunctive lithium and fluoxetine were allowed during the open-label extension. RESULTS: The mean length of olanzapine treatment was 6.6 months, with a mean modal dose of 13.9 mg/day. A significant mean improvement in the YMRS total score, baseline to endpoint (-18.01, p < .001), was observed. During treatment, 88.3% of patients experienced a remission of manic symptoms (YMRS total score < or =12), and only 25.5% subsequently relapsed (YMRS total score > or = 15). Significant improvement in HAM-D-21 scores was observed (p < .001). Forty-one percent of patients were maintained on olanzapine monotherapy. The most common treatment-emergent adverse events reported were somnolence (46.0%), depression (38.9%), and weight gain (36.3%). CONCLUSION: During up to 1 year of olanzapine therapy, either as monotherapy or in combination with lithium and/or fluoxetine, patients with bipolar disorder demonstrated significant improvement in mania and depression symptoms with a favorable safety profile. Further double-blind, controlled studies are needed to confirm these results. PMID- 11379843 TI - Optimal dosing with risperidone: updated recommendations. AB - BACKGROUND: Drug dosages utilized during controlled clinical trials are not always optimal for patients encountered in day-to-day practice. The original trials of risperidone, a novel antipsychotic, suggested that an initial target dose of 6 mg/day was appropriate, but these trials were necessarily conducted among patients who were chronically impaired, hospitalized, and often partly drug resistant. DATA SOURCES: Relevant data relating to the dosage of risperidone identified through an online (MEDLINE) search using the keywords risperidone, schizophrenia, schizoaffective disorder dementia, bipolar disorder, and dose were supplemented by a review of international and U.S. Congress abstracts in which the dose of risperidone was specifically described. CONCLUSION: On the basis of naturalistic studies, clinical audit, phase 4 trials, positron emission tomography data, and 5 years of clinical experience, the currently recommended target dose of risperidone is 4 mg/day for most patients, with less-rapid titration than previously recommended. Moreover, a lower dose than this and slower titration may be appropriate for elderly patients, young patients, and first-episode patients. PMID- 11379844 TI - An open-label study of the treatment efficacy of olanzapine for Tourette's disorder. AB - BACKGROUND: An open-label trial was performed to explore efficacy and safety of olanzapine, an atypical neuroleptic with diverse receptor activity including both dopamine-2 and serotonin-2A and -2C antagonism, for treatment of Tourette's disorder. METHOD: Ten adult patients aged 20 to 44 years with Tourette's disorder were treated using an open-label, flexible dosing schedule for 8 weeks. Three patients who continued olanzapine were reevaluated after 6 months. Three subjects were psychotropic medication naive, 5 patients experienced intolerable side effects with conventional neuroleptics, and 2 patients had remote (> or = 10 years) successful response to conventional neuroleptics. Tic severity was rated by the Yale Global Tic Severity Scale; weight, vital signs, and adverse effects were assessed weekly. Electrocardiogram, laboratory studies, and comorbid symptoms, assessed by the Yale-Brown Obsessive Compulsive Scale and ADHD Behavior Checklist for Adults, were measured at baseline and at week 8. RESULTS: Two of 10 patients prematurely discontinued olanzapine owing to excessive sedation. Of 8 patients who completed the 8-week trial, 4 (50%) demonstrated reduction of global tic severity scores by > or = 20 points, and 6 (75%) demonstrated reductions by > or = 10 points. No significant changes in comorbid symptoms were demonstrated. Sedation, weight gain, increased appetite, dry mouth, and transient asymptomatic hypoglycemia were the most common side effects. Tic improvements were maintained in 3 patients reassessed 6 months later. Final olanzapine dosages ranged from 2.5 mg to 20 mg daily (mean = 10.9 mg/day). CONCLUSION: This open-label study suggests that olanzapine should be explored as a potential alternative to conventional neuroleptic medications for treatment of motor tics and Tourette's disorder. PMID- 11379845 TI - Cost savings with intramuscular anticholinergics. PMID- 11379846 TI - Problems with ensuring a double blind. PMID- 11379848 TI - Successful treatment of nondelusional body dysmorphic disorder with olanzapine: a case report. PMID- 11379847 TI - Low-dose olanzapine for self-mutilation behavior in patients with borderline personality disorder. PMID- 11379849 TI - Olanzapine in the treatment of tardive dyskinesia: a report of 2 cases. PMID- 11379850 TI - Use of the Brief Psychiatric Rating Scale to facilitate differential diagnosis at acute inpatient admission. AB - BACKGROUND: The advent of managed care has necessitated strategies for quickly and accurately diagnosing psychiatric disorders. The aim of the present study was to ascertain whether the Brief Psychiatric Rating Scale-Anchored (BPRS-A) would be a useful adjunct to more traditional diagnostic strategies at acute inpatient admission. METHOD: Using a sample of 207 inpatients admitted during an 8-month index period, we examined the utility of the BPRS-A in predicting whether patients were more likely to be diagnosed with schizophrenia, bipolar disorder, or major depression (DSM-IV). RESULTS: Discriminant function analyses were used to correctly predict 68%, 60%, and 74% of patients diagnosed with schizophrenia, bipolar disorder, and major depression, respectively. The main predictors of diagnostic category, in descending order. were BPRS-A depressed mood item, BPRS-A positive symptoms scale, BPRS-A excitement item, BPRS-A guilt feelings item, BPRS A mannerisms and posturing item, and number of previous episodes. CONCLUSION: As efforts are directed toward continuous quality improvement within mental health settings, an emphasis must be placed on improving the efficiency and accuracy of diagnostic procedures. The BPRS-A shows promise as a time-efficient assessment instrument that may be useful in facilitating differential diagnosis at inpatient admission and may increase the likelihood that efficacious prerelease interventions and appropriate aftercare services are implemented. PMID- 11379851 TI - The preferred phosphate binder in dialysis patients. PMID- 11379852 TI - Peritoneal dialysis in patients with large body size: can it deliver adequate clearances? PMID- 11379853 TI - Management of orthostatic hypotension from autonomic dysfunction in diabetics on peritoneal dialysis. PMID- 11379854 TI - An amino acid-based peritoneal dialysis fluid buffered with bicarbonate versus glucose/bicarbonate and glucose/lactate solutions: an intraindividual randomized study. AB - OBJECTIVE: In order to study acute metabolic changes and peritoneal transport, amino acids as osmotic agent and bicarbonate as buffer were tested as new agents in peritoneal dialysis (PD) solutions. DESIGN: In a prospective, cross-over, randomized, intraindividual study, we investigated the acute metabolic changes following the application of three different PD fluids: (1) a 1% amino acid-based PD solution buffered with bicarbonate (34 mmol/L) (Amino/Bic); (2) a 1.5% glucose anhydrous-containing bicarbonate-buffered solution (34 mmol/L) (Glu/Bic); and (3) a conventional 1.5% glucose anhydrous-based dialysis solution with lactate (35 mmol/L) (Glu/Lac). SETTING: University medical center. PATIENTS: Ten nondiabetic patients stable on continuous ambulatory peritoneal dialysis (time on dialysis, 42.5 +/- 21.5 months) were treated and monitored with the test solutions over a 6 hour dwell. Three different study days followed in a randomized order for each patient (interval of 1-3 weeks). Blood and dialysate samples were taken at 0.25, 0.5, 1, 2, 4, and 6 hours. Immediately after the 1-hr dwell (and after sampling), the patients received a standardized breakfast, thereby simulating usual food intake. RESULTS: Following the application of Amino/Bic a significant increase in plasma amino acids occurred, with peak levels (maximum 250% increase) after either the 1-hr or the 2-hr dwell. Before taking the standard meal (0.5 hr, 1 hr), the mean serum glucose level with Amino/Bic was 8% +/- 13% lower than with Glu/Bic (p = 0.06) and 14% +/- 8% lower than with Glu/Lac (p < 0.01). This difference was still significant after the standard breakfast and also for the whole dwell (average serum glucose 0.5-6 hr: Amino/Bic, 91 +/- 6 mg/dL; Glu/Bic, 100 +/- 8 mg/dL; Glu/Lac, 102 +/- 7 mg/dL; p < 0.01 MANOVA). The serum insulin profiles did not differ between the fluids. A transperitoneal protein- and amino acid-related nitrogen loss of 0.49 +/- 0.18 g and 0.48 +/- 0.12 g per dwell was measured using Glu/Bic and Glu/Lac, while a positive balance of 1.80 +/- 0.43 g was achieved with Amino/Bic. The parameters of acid-base status (pH, HCO3, pCO2) remained nearly unchanged in the blood, irrespective of the solution used, while dialysate values differed markedly. No significant differences with respect to ultrafiltration (Amino/Bic, -68 +/- 199 mL/6 hr; Glu/Bic, -51 +/- 89 mL/6 hr; Glu/ Lac, -2 +/- 134 mL/6 hr) and peritoneal creatinine clearance (Amino/Bic, 4.9 +/- 0.6 mL/min; Glu/Bic, 5.1 +/- 0.6 mL/min; Glu/ Lac, 4.8 +/- 0.5 mL/min) were measured. CONCLUSIONS: Our results demonstrate that ultrafiltration and small solute clearance over a 6-hour dwell with a 1% Amino/Bic solution were comparable to those of 1.5% Glu/Bic and 1.5% Glu/Lac. Reduced serum glucose concentrations were found with Amino/Bic and this fluid compensated the transperitoneal protein nitrogen loss of about three glucose dwells. Bicarbonate buffering (34 mmol/L) did not change blood acid-base status combined with either glucose or amino acids. PMID- 11379855 TI - Gastric emptying and electrogastrography in patients on CAPD. AB - OBJECTIVE: To evaluate if impaired gastric emptying of digestible solids can explain the disturbed eating behavior in continuous ambulatory peritoneal dialysis (CAPD) patients, and if predialytic and dialytic (CAPD and hemodialysis) patients with impaired gastric emptying have a higher prevalence of electrogastrographic (EGG) abnormalities. DESIGN: Cross-sectional study. After ingestion of a 99mTc-labeled test meal, anterior and posterior 1-minute scintigraphic acquisitions were collected every 5 minutes during the first 50 minutes and thereafter every 10 minutes until 2 hours had elapsed. Simultaneously, cutaneous EGG recorded gastric myoelectric activity. SETTING: The Division of Nephrology and the Department of Nuclear Medicine at the same academic teaching hospital. PARTICIPANTS: Thirty outpatients participated in both the gastric emptying and the EGG studies. Dialysis patients should have been on dialysis for more than 3 months. For comparison, 160 healthy control subjects participated in the gastric emptying study. MAIN OUTCOME MEASURES: The following parameters were used to describe gastric emptying: lag phase 90%, half-emptying time (T50), gastric retention at 90 and 120 minutes (Ret 90/120) and gastric emptying rate (GER, %/min). Electrogastrographic measurements were expressed as percentages of normal slow waves [2.4-3.6 cycles/min (cpm)], bradygastria (1.5 2.4 cpm), and tachygastria (3.6-10 cpm). RESULTS: T50 was prolonged, Ret 90 and Ret 120 were higher, and GER was slower in male CAPD patients compared to male controls. No significant differences were found in postprandial EGG. CONCLUSION: Gastric emptying is impaired in male non-diabetic CAPD patients. However, abnormalities in postprandial EGG cannot explain this finding. PMID- 11379856 TI - Re-evaluation of solute transport groups using the peritoneal equilibration test. AB - OBJECTIVE: To determine if the previously described peritoneal equilibration test (PET)-determined solute transport groups, as defined by Twardowski, fit our patient population. DESIGN: We reviewed the 195 initial standardized PETs (on 195 patients) performed through our peritoneal dialysis program since 1989. Using the method originally defined by Twardowski using the means and standard deviations of the PET-determined dialysis/plasma ratio (D/P) of creatinine and dialysate-to 0 hour dialysate (D/D0) glucose values, transport groupings for our patient population were determined. Comparisons were then made between patient populations. RESULTS: The mean 4-hour D/P creatinine in our patients was 0.70 +/- 0.10. This compares to a mean of 0.65 +/- 0.15 as determined by Twardowski, and indicates that our patients have higher mean solute transport characteristics and tighter ranges within transport groups than previously reported. Only 2% of our patients fell into the previously described low (L) range, with 30% low average (LA), 51% high average (HA), and 17% high (H). Using our data, we would redefine the groups by a 4-hour D/P creatinine as L < 0.60, LA = 0.60-0.70, HA = 0.70 0.80, and HA > 0.80. Using these values, our population fits a Gaussian distribution with 17% L, 31% LA, 33% HA, and 19% H. CONCLUSION: Our patients have higher mean solute transport and tighter ranges within transport groups than previously reported. Using the previously defined PET-determined transport groupings, low transporters are particularly underestimated. If our population data are representative of the peritoneal dialysis population as a whole, these ranges should be redefined. PMID- 11379857 TI - Peritoneal transport of glucose in rat. AB - OBJECTIVE: To evaluate the convective transport characteristics of glucose and the effect of high glucose and insulin during experimental peritoneal dialysis in rat. METHODS: Male Sprague-Dawley rats weighing 300-400 g were used in this study. Mannitol (5%) was used as osmotic agent. Glucose was added to dialysis solution to yield a concentration of 100 mg/dL (group 1) or 300 mg/dL (group 2). Mannitol solution (5%) containing the same concentration of electrolytes and lactate but without glucose was used as control (group 3). In group 2, blood sugar was maintained at approximately 300 mg/dL by continuous intravenous infusion of 25% glucose solution and 0.9% NaCl solution. A 2-hour dwell study was performed with 30 mL of test solutions. Intraperitoneal volume was calculated by volume marker (18.5 kBq of 131I-human radioiodinated serum albumin, RISA) dilution with corrections made for the elimination of RISA from the peritoneal cavity (K(E)) and sample volume. The diffusive mass transport coefficient (K(BD)) and sieving coefficient (S(BRF)) were calculated by using the Babb-Randerson Farrell model. S was also calculated directly by using isocratic methods (S(I)). The peritoneal fluid absorption rate (K(E)) was taken into account for the calculation of S(I). RESULTS: Intraperitoneal volume was significantly higher in group 2 compared with groups 1 and 3. Peritoneal fluid absorption rate, K(E), was similar in all three groups. S(BRF) and S(I) for glucose were significantly lower in group 2 compared with groups 1 and 3. S(BRF) for glucose in group 2 was below zero and S(I) near zero. K(BD) for glucose was significantly higher in group 2 than in groups 1 and 3. Plasma and dialysate concentrations of insulin increased during the initial hour and then decreased to the baseline value in groups 1 and 3, while in group 2 it continuously increased. CONCLUSION: Significantly lower sieving coefficients for glucose in the high glucose and high insulin group suggest that transport mechanisms other than simple passive transport are involved in peritoneal glucose transport, and that high glucose per se and/or high insulin may be important factors that determine glucose transport characteristics. PMID- 11379859 TI - Serum magnesium concentration is an independent predictor of parathyroid hormone levels in peritoneal dialysis patients. AB - BACKGROUND: Parathyroid hormone (PTH) is a cardinal factor in the pathogenesis of bone disease in the dialysis population. The spectrum of renal osteodystrophy has been reported to have changed during the past years, and adynamic bone disease has emerged as the most common bone disorder in these patients. Continuous ambulatory peritoneal dialysis (CAPD) is considered a risk factor for the development of this condition, and furthermore, the adynamic bone lesion is associated with a state of relative hypoparathyroidism (hypo-PTH). Calcium, vitamin D, and phosphorus play a key role in the control of parathyroid gland function in uremic patients. However, magnesium may also be able to modulate PTH secretion in a way similar to calcium. OBJECTIVE: The aims of this study were (1) to analyze the serum Mg concentration in a large group of CAPD patients, (2) to study the relationship between serum Mg and PTH levels, and (3) to investigate whether this relationship is independent of other factors, such as calcium, phosphorus, and calcitriol, that regulate parathyroid function. PATIENTS AND METHODS: We studied 51 stable patients, aged 23-77 years, under maintenance CAPD for more than 6 months (range 8-48 months). Calcium carbonate was used as a phosphate binder in all patients, and 9 subjects also received aluminum hydroxide. No patient had been previously treated with vitamin D. Biochemical parameters were prospectively evaluated over 6 months, and the mean values were computed. RESULTS: The mean serum Mg was 1.08 +/- 0.19 mmol/L, and hypermagnesemia, defined as a Mg level higher than 1.01 mmol/L, was found in 30 patients (59%). Thirty-one subjects (60%) had an intact PTH (iPTH) level lower than 120 pg/mL and were diagnosed as having relative hypo-PTH. Except for the values of iPTH and alkaline phosphatase, the only difference between the two groups was the serum Mg concentration, which was significantly higher in patients with hypo-PTH (1.16 +/- 0.15 mmol/L vs 0.91 +/- 0.14 mmol/L; p< 0.001). Furthermore, iPTH levels were lower in patients with hypermagnesemia than in subjects with normal serum Mg (69 +/- 49 pg/mL vs 190 +/- 89 pg/mL, p < 0.001). There was a significant correlation between serum Mg and PTH levels (r= -0.70, p< 0.01). After controlling for the effect of other variables by partial correlation analysis, a significant positive association between P and PTH (r= 0.25, p < 0.05), and a negative relationship between Mg and PTH (r= -0.57, p < 0.001) were evident. A forward stepwise multiple regression analysis showed that only P and Mg predicted PTH values (multiple r = 0.59, p < 0.001). CONCLUSIONS: Hypermagnesemia and hypoparathyroidism are frequent in CAPD patients. There is a significant inverse relationship between serum Mg concentration and iPTH levels. Furthermore, this association is independent of the most important factors regulating parathyroid gland function (calcium, phosphorus, and calcitriol). These results suggest that hypermagnesemia may have a suppressive effect on PTH synthesis and/or secretion. Therefore, elevated serum Mg levels may play a role in the pathogenesis of adynamic bone disease. PMID- 11379858 TI - Surgical management of refractory exit-site/tunnel infection of Tenckhoff catheter: technical innovations of partial replantation. AB - OBJECTIVE: Catheter-related infection has been the major cause of catheter removal for peritoneal dialysis (PD) patients. A salvage technique--partial replantation of the infected catheter--was developed in our hospital to rescue catheters with refractory exit-site or tunnel infection. PATIENTS: We performed 26 partial replantations of Tenckhoff catheters for 23 patients with refractory exit-site or tunnel infection and 2 patients with near-cuff perforation of the catheter. Their problems were all resolved successfully without interruption of PD. INTERVENTIONS: We removed the infected portion of the catheter and preserved the still-functioning internal conduit, connecting it to a divided new catheter. All of the patients resumed PD immediately after the advancement of the new catheter through a new subcutaneous tunnel and exit site on the opposite side. RESULTS: No technical complications such as disconnection of the catheter or leakage of dialysate were noted. Repeated partial replantation of the catheter was done for 1 patient with a new refractory exit-site infection. Tunnel infection was not an absolute contraindication for this procedure. About one third (34.6%) of our patients had preoperative tunnel infection. CONCLUSION: Partial replantation of a Tenckhoff catheter is a simple and effective procedure for patients with refractory exit-site/tunnel infection and patients with near cuff perforation of the catheter. Repeated partial replantation is also feasible for repeat exit-site infections. PMID- 11379860 TI - Effects of amino acid dialysis solution on the nutrition of continuous ambulatory peritoneal dialysis patients. AB - OBJECTIVE: To evaluate the influence of 1.1% amino acid dialysis solution (AADS) on parameters of nutrition in continuous ambulatory peritoneal dialysis (CAPD) patients. STUDY DESIGN: Studies were performed in 8 men, using AADS for the overnight exchange. Before starting AADS, food intake, nutritional status, and laboratory indices were evaluated and compared to the respective parameters obtained after 3 and 6 months of treatment with AADS, as well as after 3 months of AADS withdrawal. With the start of AADS, doses of antacids were increased and modified during AADS administration; the modified doses were continued through 3 months after cessation of AADS. Another group of CAPD patients using standard dialysis solutions served as controls. In these patients the same parameters were evaluated four times at 3-month intervals. RESULTS: Administration of AADS resulted in: (1) 91% absorption of amino acids and improvement of serum amino acid pattern; (2) no change in nutritional intake during the treatment, but after the 3 months of AADS therapy, levels of nutrient intake were lower than those 3 months after withdrawal of AADS with correction of metabolic acidosis; (3) no change in indices of nutritional status, but 3 months after AADS discontinuation, total body weight, lean body mass, and body mass index were significantly higher than those shown after 3 months of treatment; (4) an increase in hemoglobin concentration, hematocrit, BUN, and blood H+. The examined parameters were not significantly changed in patients treated for 9 months with standard dialysis solutions exclusively. The values of nitrogen balance obtained during AADS administration and after 3 months of AADS withdrawal were significantly higher than those obtained in the respective periods in the control group. The blood pH, pCO2, and HCO3- in the last period of the study were higher in the AADS group than in the control group. CONCLUSION: In relatively well-nourished CAPD patients, overnight AADS administration results in increased serum concentration of amino acids without changes in other nutritional parameters. The use of AADS should be associated with increased doses of antacid medication, which abolishes the metabolic effects of acidosis that develop during AADS administration and facilitates positive nitrogen balance. PMID- 11379861 TI - Validity of a standard information protocol provided to end-stage renal disease patients and its effect on treatment selection. AB - OBJECTIVE: To evaluate the validity of a standard information package, comprising written and audiovisual aids, for end-stage renal disease (ESRD) patients in a predialysis program. STUDY DESIGN: A multicenter study comprising patients entering a predialysis program. Three questionnaires were developed to gather data in this study: (1) a pre-information package questionnaire that evaluates the patient's initial knowledge of ESRD and the treatment options available (pre informed patients); (2) a post-information package questionnaire that evaluates the patient's knowledge of ESRD and treatment options after being informed according to the protocol (post-informed patients); (3) a "start of the treatment" questionnaire that deals with the patient's choice of treatment at the time of starting dialysis, as well as the reasons leading to that choice. In all three questionnaires the patient's age, gender, level of creatinine clearance (Ccr), and hematocrit were recorded. INCLUSION CRITERIA: Any patient who was on a predialysis program in the participating centers. RESULTS: Three hundred and four patients were evaluated across 14 participating centers. Initial knowledge was assessed in 216 pre-informed patients (questionnaire 1). Patients were then guided through the information package. One hundred and fifty-eight patients answered the post-information package (questionnaire 2). During the course of the study, 174 patients (of the initial 304) started renal replacement therapy. Of these, 49.4% (86 patients) had received predialysis information according to our study protocol. All the patients who received the information throughout the trial improved their knowledge of ESRD and treatment options; this improvement was statistically significant. CONCLUSIONS: The treatment options least well known at the start of the study were the peritoneal dialysis techniques. After receiving the information package, patients had an equal knowledge of all the different treatments, although hemodialysis was still the most familiar. This improvement in knowledge enabled patients who started a dialysis treatment to choose a therapy according to their own preferences. Their selections were as follows: 44% of the patients chose hemodialysis, 40% chose continuous ambulatory peritoneal dialysis, and 16% chose automated peritoneal dialysis. The standard information package, used as a patient education program, effectively resulted in patients having a significantly improved level of knowledge and understanding of ESRD and the different treatment options available. PMID- 11379862 TI - Sclerosing encapsulating peritonitis: a case successfully treated with immunosuppression. PMID- 11379863 TI - Intraperitoneal fluconazole for fungal peritonitis in CAPD: report of two cases. PMID- 11379864 TI - Methicillin resistance patterns associated with peritonitis in a university-based peritoneal dialysis center. PMID- 11379865 TI - Effect of erythropoietin therapy on serum apolipoprotein A1 levels in patients undergoing chronic peritoneal dialysis. PMID- 11379866 TI - Recurrent sterile peritonitis at onset of treatment with icodextrin solution. PMID- 11379867 TI - Bowel perforation due to hypotension in an infant with prune belly syndrome undergoing peritoneal dialysis. PMID- 11379868 TI - A new laparoscopic technique for CAPD catheter placement. PMID- 11379869 TI - Intraperitoneal amino acids in CAPD: an 18 month experience. PMID- 11379870 TI - Nursing application: peritoneal dialysis nursing in the United Kingdom--impact of the nurse and U.K. PD trends. PMID- 11379871 TI - Literature: September-October 1999. PMID- 11379872 TI - Identification of a new point mutation in the human xanthine dehydrogenase gene responsible for a case of classical type I xanthinuria. AB - A 60-year-old Japanese man was diagnosed as having hypouricemia at an annual health check-up. The routine laboratory data was not remarkable except that the patient's hypouricemia and plasma levels of xanthine and hypoxanthine were much higher than those of normal subjects. Furthermore, the patient's daily urinary excretion of xanthine and hypoxanthine was markedly increased compared with reference values. The xanthine dehyrogenase activity of the duodenal mucosa was below the limits of detection. Nevertheless, allopurinol was metabolized to oxypurinol in vivo. Based on these findings, a subtype of classical xanthinuria (type I) was diagnosed. The xanthine dehyrogenase protein was detected by Western blotting analysis. Sequencing of the cDNA of the xanthine dehyrogenase obtained from the duodenal mucosa revealed that a point mutation of C to T had occurred in nucleotide 445. This changed codon 149 from CGC (Arg) to TGC (Cys), a finding that has not been previously reported in patients with classical xanthinuria type I. PMID- 11379873 TI - Hearing impairment in patients with 3243A-->G mtDNA mutation: phenotype and rate of progression. AB - The relationship between the phenotype and the genotype is complex in diseases caused by mutations in mitochondrial DNA (mtDNA). The 3243A-->G mutation in mtDNA frequently leads to sensorineural hearing impairment (HI), a phenotype that can be assessed in severity by audiometry; hence, consecutive audiograms can give an estimate of the rate of HI progression. We examined the audiological phenotype of 38 patients (14 men, 24 women; mean age: 45+/-14 years) who possessed the 3243A- >G mutation and who belonged to a population-based cohort ascertained in the province of Northern Ostrobothnia, Finland. The subjects took part in an otorhinolaryngologic examination, including audiometry. Factors modulating the severity of HI were analyzed, and the rate of HI progression was calculated. The better ear hearing level (BEHL) at frequencies 0.5, 1, 2, and 4 kHz (BEHL0.5 4kHz) was greater than 20 dB suggesting HI in 28 patients (74%). A good correlation (r=0.428, P=0.009) was found between BEHL0.5-4kHz and the degree of the mutant heteroplasmy. BEHL0.5-4kHz was worse in men than in women, and women outnumbered men among patients with normal hearing or mild HI. In addition, 181 consecutive audiograms were reviewed from 24 patients with HI. The rate of HI progression was calculated to be 2.9 dB/year in men and 1.5 dB/year in women, being clearly faster than the rates that have been observed in the corresponding age group in the general population. A high degree of mutant heteroplasmy, male gender, and age were found to increase the severity of HI. Phenotypic difference by gender may thus be a more universal phenomenon in mitochondrial diseases, not only being associated with Leber's hereditary optic neuropathy. This study provides the first estimate of the rate of disease progression among patients with the 3243A-->G mutation. PMID- 11379874 TI - Complete and rapid scanning of the cystic fibrosis transmembrane conductance regulator (CFTR) gene by denaturing high-performance liquid chromatography (D HPLC): major implications for genetic counselling. AB - More than 900 mutations and more than 200 different polymorphisms have now been reported in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Ten years after the cloning of the CFTR gene, the complete scanning of the 27 exons to identify known and novel mutations remains challenging. Rapid accurate identification of mutated alleles is important for prenatal diagnosis, for cascade screening in families at risk of cystic fibrosis (CF) and for understanding the correlation between genotype and phenotype. In this study, we report the successful use of denaturing ion-pair reverse-phase high performance liquid chromatography (D-HPLC) to analyse rapidly the complete coding sequence of the CFTR gene. With 27 pairs of polymerase chain reaction primers, we optimised the temperature conditions required for the analysis of each amplicon and validated thetest conditions on samples from a panel of 1552 CF patients who came from France and other European countries and who had mutations and polymorphisms located in the various melting domains of the gene. D-HPLC identified 415 mutated alleles previously characterised by denaturing gradient gel electrophoresis and DNA sequencing, plus 74 novel mutations reported here. This new technique for screening DNA for sequence variation was extremely accurate (it identified 100% of the CFTR alleles tested so far) and rapid (the complete CFTR gene could be analysed in less than a week). Our approach should reduce the number of untyped CF alleles in populations and thus decrease the residual risk in couples at risk of CF. This technique may be important not only for CF,but also for many other genes with a high frequency of point mutations at a variety of sites. PMID- 11379875 TI - Identification of novel DKC1 mutations in patients with dyskeratosis congenita: implications for pathophysiology and diagnosis. AB - Dyskeratosis congenita (DC) is characterised by the failure of those tissues that are rapidly dividing in the adult, particularly the skin and haemopoietic system. The X-linked form of the disease is caused by mutations in the DKC1 gene. To date the only DKC1 mutations detected result in alterations in the amino acid sequence of dyskerin. Dyskerin is the catalytic subunit of the H+ACA box small nucleolar RNA particles responsible for the site-specific pseudouridination of rRNA and in humans is also a component of the telomerase complex. In order to further characterise the disease at the molecular level, male DC patients from 25 families were screened for mutations in the DKC1 gene. Sequence variations were detected in 10 of these families. In five families, previously identified mutations were detected. Of the five novel sequence changes, three were coding changes: R158 W, S280R and P384L. A fourth sequence change was detected in the 5' flanking region that disrupts a putative Spl transcription factor binding site. An intronic change was also detected that resulted in the partial incorporation of a portion of intron 1 into the mRNA. The identification of this mutation highlights the importance of screening for mutations that cause the partial aberrant splicing of mRNA. This is the first report of DKC1 mutations that are predicted to affect the level of expression of dyskerin. This suggests that a decrease in the amount of the normal protein may cause the disease. PMID- 11379876 TI - Mutation analysis of the origin recognition complex subunit 5 (ORC5L) gene in adult patients with myeloid leukemias exhibiting deletions of chromosome band 7q22. AB - The ORC5L gene encoding a subunit of the human origin recognition complex (ORC) maps to chromosome band 7q22, a region frequently deleted in adult acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS). Because of its localization within a region that is commonly deleted in patients with myeloid malignancies and because of the implication of its protein product in cell cycle control (DNA replication) and regulation of gene expression (transcriptional silencing), ORC5L appeared to be a candidate tumor suppressor gene for myeloid disorders associated with 7q22 deletions. Polymerase chain reaction amplification and sequencing analysis of the coding region of the remaining ORC5L allele has not revealed any mutations in nine patients with AML or MDS exhibiting 7q22 deletions. Allelic expression analysis indicates that ORC5L is not imprinted. These data suggest that ORC5L does not function as a tumor suppressor in patients with myeloid neoplasms. PMID- 11379877 TI - Haplotype analysis of genomic polymorphisms in and around the myotonic dystrophy locus in diverse populations of India. AB - The frequencies of haplotypes based upon the (CTG)n repeat and three other biallelic markers in and around the myotonic dystrophy (DM) locus were estimated in 13 ethnically, linguistically and geographically diverse sub-populations of India. The range of CTG repeats in caste populations was 5-31, while in tribal populations the range was shorter (5-23). Extensive variation in frequencies of large (CTG)n alleles (> or =18 repeats) was found in Indian populations. The implications of this finding on DM epidemiology are discussed. Haplotype diversity was found to be very high in most populations. The majority of the Indian DM patients carried a haplotype that is commonly found among DM patients globally; this is the most common haplotype in the class of large (> or =18 repeats) CTG alleles. However, one haplotype was found to be present in particularly high frequency in Indian populations; this haplotype was also found among Indian DM patients. This haplotype may be a characteristic of Indian and possibly of other East Asian populations. PMID- 11379878 TI - Phenotypic effects of balanced X-autosome translocations in females: a retrospective survey of 104 cases reported from UK laboratories. AB - Females with balanced X-autosome translocations are a clinically heterogeneous group of patients in which X breakpoint position and replication behaviour may influence phenotypic outcome. This study reviewed all cases reported by UK cytogenetics laboratories over a 15-year period (1983-1997). Publication bias was avoided by reviewing all reported cases. One hundred and four female carriers were identified, 62 of who were probands. By reason for referral, these were: multiple congenital abnormalities and/or developmental delay (MCA/DD): 26 (42%); gonadal dysfunction: 22 (35%); phenotypically normal with or without recurrent miscarriage (NRM): 9 (15%); recognized X-linked syndrome: 5 (8%). The information obtained was compared with published data and with data from the authors' own laboratories of female patients with balanced autosome-autosome translocations (n=115). We concluded that: (1) MCA/DD cases were significantly over-represented compared to previous published data (P<0.005) and were more common than in female probands with balanced autosome-autosome translocations (P<0.05). (2) MCA/DD cases showed random breakpoint distribution along the X chromosome (P>0.05). MCA/DD cases with subtelomeric breakpoints at Xp22 or Xq28 were not always associated with deviation from the expected pattern of X-inactivation where this was known. De novo cases were significantly more likely to be assigned as MCA/DD than any other category (P<0.005). (3) Gonadal dysfunction (GD) was invariably associated with a 'critical region' breakpoint, Xq13-q26, (20/22 probands). However, 7/44 (16%) of patients surveyed had breakpoints within Xq13-Xq26 and proven fertility. (4) Recognized 'X-linked syndrome' cases were significantly under-represented (P<0.001) compared to previous published data. PMID- 11379879 TI - Segregation of a mutation in CNGB1 encoding the beta-subunit of the rod cGMP gated channel in a family with autosomal recessive retinitis pigmentosa. AB - Retinitis pigmentosa (RP) is a clinically and genetically heterogeneous group of retinal diseases leading to blindness. By performing full genome linkage analysis in a consanguineous French family affected with severe autosomal recessive RP, we have excluded linkage to known loci involved in RP and mapped a novel locus to chromosome 16q13-q21 (Zmax=2.83 at theta=0 at the D16S3089 locus). Two candidate genes KIFC3 and CNGB1 mapping to this critical interval have been screened for mutations. The CNGB1 gene, which encodes the beta-subunit of the rod cGMP-gated channel, is mutated in the family presented in this study. PMID- 11379881 TI - Late-onset Stargardt disease is associated with missense mutations that map outside known functional regions of ABCR (ABCA4). AB - Based on recent studies of the photoreceptor-specific ABC transporter gene ABCR (ABCA4) in Stargardt disease (STGD1) and other retinal dystrophies, we and others have developed a model in which the severity of retinal disease correlates inversely with residual ABCR activity. This model predicts that patients with late-onset STGDI may retain partial ABCR activity attributable to mild missense alleles. To test this hypothesis, we used late-onset STGDI patients (onset: > or =35 years) to provide an in vivo functional analysis of various combinations of mutant alleles. We sequenced directly the entire coding region of ABCR and detected mutations in 33/50 (66%) disease chromosomes, but surprisingly, 11/33 (33%) were truncating alleles. Importantly, all 22 missense mutations were located outside the known functional domains of ABCR (ATP-binding or transmembrane), whereas in our general cohort of STGDI subjects, alterations occurred with equal frequency across the entire protein. We suggest that these missense mutations in regions of unknown function are milder alleles and more susceptible to modifier effects. Thus, we have corroborated a prediction from the model of ABCR pathogenicity that (1) one mutant ABCR allele is always missense in late-onset STGD1 patients, and (2) the age-of-onset is correlated with the amount of ABCR activity of this allele. In addition, we report three new pseudodominant families that now comprise eight of 178 outbred STGD1 families and suggest a carrier frequency of STGD1-associated ABCR mutations of about 4.5% (approximately 1/22). PMID- 11379880 TI - Linkage of prostate cancer susceptibility loci to chromosome 1. AB - Three prostate cancer susceptibility genes have been reported to be linked to different regions on chromosome 1: HPC1 at 1q24-25, PCAP at 1q42-43, and CAPB at 1p36. Replication studies analyzing each of these regions have yielded inconsistent results. To evaluate linkage across this chromosome systematically, we performed multipoint linkage analyses with 50 microsatellite markers spanning chromosome 1 in 159 hereditary prostate cancer families (HPC), including 79 families analyzed in the original report describing HPC1 linkage. The highest lod scores for the complete dataset of 159 families were observed at 1q24-25 at which the parametric lod score assuming heterogeneity (hlod) was 2.54 (P=0.0006) with an allele sharing lod of 2.34 (P=0.001) at marker D1S413, although only weak evidence was observed in the 80 families not previously analyzed for this region (hlod=0.44, P=0.14, and allele sharing lod=0.67, P=0.08). In the complete data set, the evidence for linkage across this region was very broad, with allele sharing lod scores greater than 0.5 extending approximately 100 cM from 1p13 to 1q32, possibly indicating the presence of multiple susceptibility genes. Elsewhere on chromosome 1, some evidence of linkage was observed at 1q42-43, with a peak allele sharing lod of 0.56 (P=0.11) and hlod of 0.24 (P=0.25) at D1S235. For analysis of the CAPB locus at 1p36, we focused on six HPC families in our collection with a history of primary brain cancer; four of these families had positive linkage results at 1p36, with a peak allele sharing lod of 0.61 (P=0.09) and hlod of 0.39 (P=0.16) at D1S407 in all six families. These results are consistent with the heterogeneous nature of hereditary prostate cancer, and the existence of multiple loci on chromosome 1 for this disease. PMID- 11379882 TI - Characterisation of novel point mutations in the survival motor neuron gene SMN, in three patients with SMA. AB - We report two novel mutations in three cases of spinal muscular atrophy (SMA), including two distant cousins who followed an unexpectedly severe course. Diagnosis was confirmed by reduced SMN protein and full-length SMN mRNA levels. Sequencing of the non-deleted SMN1 gene revealed a single G insertion at the end of exon 1 in the two cousins and a novel G275S exon 6 missense mutation in the milder case. PMID- 11379883 TI - Fusing of dental ceramics to titanium. AB - Titanium is known as a useful biometal because of its good biocompatibility and mechanical performance. However, titanium is chemically an exceptional metal, reacting strongly with gaseous elements like oxygen, hydrogen, and nitrogen and also dissolving them extensively. This high reactivity causes problems, for example, when dental ceramics are fused to titanium. Commercial ceramic-titanium systems are increasingly used in prosthetic dentistry, but little is known about the microstructure and composition of the system. Better understanding of chemical reactions between ceramics and titanium is necessary if mechanically more compatible ceramic-titanium bonds are to be developed. This review deals with titanium as a metal, titanium's affinity for nonmetallic elements (especially oxygen), and reactions with other elements. Different aspects are discussed relative to the fusing of dental ceramics to titanium. PMID- 11379884 TI - Adhesion of N-methacryloyl-omega-amino acid primers to collagen analyzed by 13C NMR. AB - Previously, we reported that the strength of the interaction between N methacryloyl-omega-amino acid (NMomegaA) primers and dentinal collagen exhibited a strong correlation with the bond strength of the resin to etched dentin. To determine the pertinent functional groups of the amino acid residues in the dentinal collagen, to which the amide and/or the carboxylic acid groups of the NMomegaAs are adsorbed, we used 13C NMR techniques--primarily through the observation of spin-lattice relaxation times, Ti--to investigate the adsorption characteristics resulting from the interaction of NMomegaAs with a model oligopeptide for collagen, (PPG)5. The addition of NMomegaAs to a collagenous solution resulted in a decrease in the T1 values of the carbonyl carbons attributed to the carboxylic acid of the C-terminal Gly and to the third amide of the N-terminal Pro residues in the (PPG)5 molecule, thus reflecting the formation of hydrogen-bonded interactions. PMID- 11379885 TI - A new Ti-5Ag alloy for customized prostheses by three-dimensional printing (3DP). AB - Three important considerations in the fabrication of customized cranio maxillofacial prostheses are geometric precision, material strength, and biocompatibility. Three-dimensional printing (3DP) is a rapid part-fabrication process that can produce complex parts with high precision. The aim of this study was to design, synthesize by 3DP, and characterize a new Ti-5Ag (wt%) alloy. Silver nitrate was found to be an appropriate inorganic binder for the Ti powder based skeleton, and the optimum sintering parameters for full densification were determined. The hardness of the Ti-5Ag alloy was shown to be much higher than that of a pure titanium sample. Potentiodynamic measurements, carried out in saline solution at body temperature, showed that the Ti-5Ag alloy had good passivation behavior, similar to that of pure titanium. It is concluded that the Ti-Ag system may be suitable for fabrication of customized prostheses by 3DP. PMID- 11379886 TI - Properties of opaque resin composite containing coated and silanized titanium dioxide. AB - Titanium dioxide (TiO2) is mainly used as a pigment in opaque resin composites for application to the surface of a metal framework. The hypothesis in this paper is that particles of silica/alumina (SiO2/Al2O3)-coated TiO2 treated with a silane coupling agent could bond effectively with resin monomers of opaque resin composites. Untreated TiO2 was used as the control filler. Compressive and flexural strength specimens were prepared by the heat-curing method, because these bulk specimens could not be made by the typical photo-curing method. The treated composite had significantly higher compressive and flexural strengths than the untreated composite after 6 months' immersion in water. Scanning electron microscopy of the fractured composite surfaces showed an interface failure between TiO2 and resin for the untreated composite and cohesive failure within the resin for the treated composite after 6 months' immersion. The light activated opaque resin composite containing treated TiO2 exhibited significantly higher bond strength to a noble dental alloy after 5000 thermal cycles than that containing untreated TiO2. Thus, silanized SiO2/Al2O3-coated TiO2 appears to be clinically useful as a filler of opaque resin composites. PMID- 11379887 TI - Metabolic effects of dental resin components in vitro detected by NMR spectroscopy. AB - Earlier studies have shown that the comonomer triethyleneglycol-dimethacrylate (TEGDMA) and the photostabilizer 2-hydroxy-4-methoxybenzophenone (HMBP) are cytotoxic and inhibit cell growth. It was the aim of this study to elucidate the underlying metabolic effects of TEGDMA and HMBP on immortal contact-inhibited Swiss albino mouse embryo cells (3T3 fibroblasts) by nuclear magnetic resonance (NMR) spectroscopy. Cell extracts and culture media were analyzed by NMR spectroscopy for metabolic changes after incubation for 24 hours with ED20 concentrations of TEGDMA and HMBP. TEGDMA could be detected in all fractions (cytosol, lipid fractions, and culture media) of 3T3 cells, while HMBP was found only in the lipid fraction accumulated at a maximum rate (51 nmol/mg DNA) compared with TEGDMA (27 nmol/mg DNA). TEGDMA increased the concentration of phosphomonoesters to 180+/-36% and decreased the phosphodiesters to 65+/-5% of controls (control = 100%). Thus, the turnover of phospholipids was enhanced, whereas content and composition of phospholipids of membranes did not alter markedly. Additionally, TEGDMA changed the metabolic state of cells, indicated by slight decreases of nucleoside triphosphates and an increase in the ratio of nucleoside diphosphates to nucleoside triphosphates, while HMBP had no effect. The most remarkable effect of TEGDMA was a nearly complete decline of the intracellular glutathione levels. Analysis of our data shows that NMR spectroscopy of cell-material interactions may reveal metabolic effects of organic test substances which are not detectable by standard in vitro assays. The comonomer TEGDMA affected the metabolism of the cells on different levels, while HMBP accumulated in the lipid fraction and induced significantly fewer effects on cell metabolism. PMID- 11379888 TI - Dynamic properties of the human temporomandibular joint disc. AB - The cartilaginous intra-articular disc of the human temporomandibular joint shows clear anteroposterior variations in its morphology. However, anteroposterior variations in its tissue behavior have not been investigated thoroughly. To test the hypothesis that the mechanical properties of fresh human temporomandibular joint discs vary in anteroposterior direction, we performed dynamic indentation tests at three anteroposteriorly different locations. The disc showed strong viscoelastic behavior dependent on the amplitude and frequency of the indentation, the location, and time. The resistance against deformations and the shock absorbing capabilities were larger in the intermediate zone than in regions located more anteriorly and posteriorly. Because several studies have predicted that the intermediate zone is the predominantly loaded region of the disc, it can be concluded that the topological variations in its tissue behavior enable the disc to combine the functions of load distribution and shock absorption effectively. PMID- 11379889 TI - Mitogenic effects of neutrophins on a periodontal ligament cell line. AB - Nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) are three representative neurotrophins responsible for the differentiation and survival of neurons, and their high-affinity receptors are tropomyosin-receptor-kinase (TRK)A, TRKB, and TRKC, respectively. In this study, we investigated the expression of neurotrophins in a mouse periodontal ligament cell line (MPL), by reverse transcription-polymerase chain-reaction (RT-PCR) and enzyme-linked immunoabsorbent assay (ELISA). We also studied the expression of TRK receptors on MPL by immunostaining and the effects of neurotrophins on the proliferation of MPL, with a hypothesis of autocrine mechanism of neurotrophins. Each neurotrophin and TRK receptor was expressed, and neurotrophins enhanced the proliferation of MPL. These findings suggest that the MPL has functional neurotrophin receptors involved in an autocrine function of neurotrophins. The expression level of neurotrophins and TRKs showed the reverse pattern, and we propose an auto-regulatory mechanism of ligands and receptors in accordance with the level of synthesized neurotrophins. PMID- 11379892 TI - Cloning and characterization of the mouse and human enamelin genes. AB - Enamelin is likely to be essential for proper dental enamel formation. It is secreted by ameloblasts throughout the secretory stage and can readily be isolated from the enamel matrix of developing teeth. The gene encoding human enamelin is located on the long arm of chromosome 4, in a region previously linked to an autosomal-dominant form of amelogenesis imperfecta (AI). To gain information on the structure of the enamelin gene and to facilitate the future assessment of the role of enamelin in normal and diseased enamel formation, we have cloned and characterized the mouse and human enamelin genes. Both genes are about 25 kilobases long. The enamelin gene has 10 exons interrupted by 9 introns. Translation initiates in exon 3 and terminates in exon 10. All of the intron/exon junctions within the mouse and human enamelin coding regions are between codons, so there are no partial codons in any exon, and deletion of one or more coding exons by alternative RNA splicing would not shift the downstream reading frame. PMID- 11379890 TI - Dual regulation of osteoclast differentiation by periodontal ligament cells through RANKL stimulation and OPG inhibition. AB - Periodontal ligament (PDL) cells play an important role in maintaining the homeostasis of periodontal tissues. However, it is not known how PDL cells contribute to osteoclastogenesis. In this study, we examined the consequences of cell-to-cell interactions between peripheral blood mononuclear cells (PBMCs) and PDL cells during osteoclastogenesis. PBMCs were co-cultured directly or indirectly with PDL cells for two to four weeks. PBMCs that were directly co cultured with PDL cells formed significantly more resorption pits on dentin slices than did PBMCs that were cultured alone. However, soluble factor(s) produced from PDL cells inhibited the formation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells. Furthermore, PDL cells expressed both receptor activator nuclear factor kappa B ligand (RANKL) and osteoprotegerin (OPG) mRNA. In conclusion, PDL cells support osteoclastogenesis through cell-to-cell contact. PDL cells might regulate osteoclastogenesis by opposing mechanisms--stimulation of resorptive activity by RANKL and inhibition by OPG--thus affecting processes such as periodontitis and orthodontic tooth movement. PMID- 11379891 TI - Gene transfer and expression of platelet-derived growth factors modulate periodontal cellular activity. AB - Platelet-derived growth factor (PDGF) is a potent stimulator of wound healing. PDGF gene therapy may promote greater periodontal regeneration than local protein application, due to sustained growth factor delivery to the target tissue. This investigation tested the ability of recombinant adenoviruses (rAds) encoding PDGF A or PDGF-1308 (a PDGF-A dominant-negative mutant that disrupts endogenous PDGF bioactivity) to affect cells derived from the periodontium. Osteoblasts, periodontal ligament fibroblasts, and gingival fibroblasts were transduced with rAds, and gene expression, DNA synthesis, and cell proliferation were evaluated. The results revealed strong message for the PDGF-A gene for 7 days following gene delivery. Ad2/PDGF-A enhanced the mitogenic and proliferative response in all cell types, while Ad2/PDGF-1308 potently inhibited mitogenesis and proliferation. In conclusion, Ad2/PDGF can effectively transduce cells derived from the periodontium and promote biological activity equivalent to PDGF-AA. These studies support the potential use of gene therapy for sustained PDGF release in periodontal tissues. PMID- 11379893 TI - Antifungal resistance of candidal biofilms formed on denture acrylic in vitro. AB - Denture biofilms represent a protective reservoir for oral microbes. The study of the biology of Candida in these biofilms requires a reliable model. A reproducible model of C. albicans denture biofilm was developed and used to determine the susceptibility of two clinically relevant C. albicans isolates against 4 antifungals. C. albicans, growing as a biofilm, exhibited resistance to amphotericin B, nystatin, chlorhexidine, and fluconazole, with 50% reduction in metabolic activity (50% RMA) at concentrations of 8, 16, 128, and > 64 microg/mL, respectively. In contrast, planktonically cultured C. albicans were susceptible (50% RMA for the same antifungals was obtained at 0.25, 1.0, 4.0, and 0.5 microg/mL, respectively). In conclusion, results obtained by means of our biofilm model show that biofilm-associated C. albicans cells, compared with cells grown in planktonic form, are resistant to antifungals used to treat denture stomatitis. PMID- 11379894 TI - Chewing stimulates secretion of human salivary secretory immunoglobulin A. AB - Immunoglobulin A (IgA) is the most abundant immunoglobulin in saliva and other mucosal secretions and plays an important role in mucosal immunity. The present study examined whether secretion of IgA, like other salivary proteins, is increased by reflex stimulation. Parotid saliva was collected from subjects into separate vials under resting conditions and during chewing-stimulated secretion over 45 min. Enzyme-linked immunosorbent assay (ELISA) indicated that chewing increased IgA secretion. The extent and pattern of the increase were similar to those of total protein and acinar cell amylase. SDS gel electrophoresis and Western blotting showed that high-molecular-weight forms of IgA-containing secretory component predominated in all saliva samples. Secretory component, the cleaved epithelial receptor for polymeric IgA, was secreted in a pattern very similar to that of IgA. It is concluded that chewing stimulates epithelial cell transcytosis of IgA and increases secretion of secretory IgA into saliva. PMID- 11379895 TI - Increased frequency of FcgammaRIIIb-NA1 allele in periodontitis-resistant subjects in an elderly Japanese population. AB - Many elderly people show minimum periodontal tissue destruction, which might be partly due to genetic advantages in host immune response against periodontopathic bacteria. The human IgG Fc receptor IIIb on neutrophils bears a NA1-NA2 polymorphism. The FcgammaRIIIb-NA1 displays a more efficient interaction with IgG1- and IgG3-opsonized bacteria, compared with the FcgammaRIIIb-NA2. We investigated a 70-year-old Japanese population (n = 599) to determine whether the FcgammaRIIIb polymorphism was associated with resistance to periodontitis. Among subjects with > or = 20 teeth present, periodontitis-resistant (n = 46) and periodontitis-susceptible groups (n = 73) were selected based on the percentage of sites with > or = 4 mm probing attachment loss in the entire dentition. The FcgammaRIIIb-NA1 allotype was overrepresented in the periodontitis-resistant group, compared with the periodontitis-susceptible group (chi2 = 4.89, p = 0.03, odds ratio = 1.87, 95% CI, 1.07 to 3.28). This suggests that FcgammaRIIIb-NA1 may be associated with resistance to periodontitis. PMID- 11379896 TI - A novel mutation in the keratin 13 gene causing oral white sponge nevus. AB - White sponge nevus (WSN) is an autosomal-dominantly inherited form of mucosal leukokeratosis. Defects in keratins, proteins that form the stress-bearing cytoskeleton in epithelia, have been shown to cause several epithelial fragility disorders. Recently, mutations in the genes encoding mucosal-specific keratins K4 and K13 were shown to be the underlying cause of WSN. We have studied a large Scottish family with 19 persons affected by WSN in four generations. The K4 locus was excluded by genetic linkage analysis; however, genetic linkage consistent with a K13 defect was obtained. Subsequently, a heterozygous missense mutation 335A>G was detected in exon 1 of the KRT13 gene, predicting the amino acid change N112S in the 1A domain of the K13 polypeptide. The mutation was confirmed in affected family members and was excluded from 50 unaffected people by restriction enzyme analysis. These results confirm that mucosal keratin defects are the cause of WSN. PMID- 11379897 TI - Randomized controlled evaluation of non-surgical treatments for temporomandibular joint anterior disk displacement without reduction. AB - The common methods for treating anterior disk displacement without reduction (ADDwor) are not based on randomized controlled clinical trials. Our study evaluated non-surgical treatments in 69 MRI-confirmed ADDwor subjects (m/f = 6/63). Subjects were randomly assigned to a control group and one of two treatment groups. Outcomes included maximum mouth opening, visual analogue scale of pain, and daily activity limitation. Calibrated examiners collected data at the initial interview and at 0, 2, 4, and 8 weeks of treatment. At the eight-week point, within-group improvements were present for all variables, for all groups. Between-group differences were not highly evident, with only mean daily activity limitation for the self-care/NSAID group being significantly lower than that of the occlusal appliance/jaw mobilization + self-care/NSAID group at the two- and four-week time-points. These results suggest that ADDwor subjects will improve with only minimal treatment intervention, and no significant difference was evident for the treatments tested and the control condition. PMID- 11379898 TI - Building materials radon exhalation rate: ERRICCA intercomparison exercise results. AB - The Nuclear Engineering Section of the National Technical University of Athens undertook the organisation of a European building material radon exhalation rate intercomparison exercise in the framework of the European Research into Radon In Construction Concerted Action (ERRICCA). The intercomparison started in June 1998 and it was concluded in February 1999. Twenty participants from 13 countries took part. The exercise focused on the radon exhalation rate determination from a concrete slab, specially constructed to produce radon surface flux well below 10 mBqm(-2) s(-1). This paper describes the measurement results obtained using different instruments and methods in order to assess 'state-of-the-art' low-level radon exhalation measurements, being performed around Europe. Results are compared to each other and they provide an indication of the collective precision of such measurements for low exhalation rates. The agreement, with a few exceptions, is satisfactorily good. PMID- 11379899 TI - Radioactivity in building materials: room model analysis and experimental methods. AB - First, models (room models) published in the international literature allowing the exposure to gamma radiation indoors due to building materials to be assessed are reviewed and discussed. For one of them, a sensitivity analysis concerning the effect of changing the parameters (e.g. dimensions of the room, thickness and density of the walls, etc.) used in calculations is performed. Second, a method is proposed for calculating the activity concentration in the walls of a room using: (a) the measured absorbed dose rate in air in the room; (b) the contributions of natural radionuclides (238U, 232Th and 40K) to the absorbed dose rate in air assessed by means of gamma spectrometry indoors; and (c) the specific dose rate (nGy h(-1) per Bq kg(-1)) of natural radionuclides, for the chosen room geometry, calculated with the room model. PMID- 11379900 TI - Laboratory assessment of flexible thin-film membranes as a passive barrier to radon gas diffusion. AB - This paper presents the experimental results of utilizing a flexible thin-film membrane as a passive barrier to radon gas diffusion. Nine commercially available membranes of various compositions and thicknesses were evaluated as retardant to radon gas diffusion. The radon gas concentration ratios across the thin-film membranes alone and in combination with an adjacent concrete sample (effective diffusion coefficient) were measured in a laboratory system with state-of-the-art instrumentation. An 8.89-cm diameter, 10.2-cm thick concrete sample of standard composition (w/c = 0.5 and cement:sand:gravel = 1:2:4) was used to simulate a basement and slab-on-grade foundation typical of Wisconsin. The radon gas transport characteristics of this concrete sample (porosity, permeability and diffusion) are documented. The experimentation has identified two superior flexible thin-film membranes that may be employed as effective barriers to radon gas diffusion. These include: Polyethylene Naphthalate (7.62 x 10(-5) m) and Polyethylene Terephthalate Glycol, PETG (7.62 x 10(-5) and 1.27 x 10(-4) m) which had average diffusion coefficients, D, of 4.10 x 10(-14) and 1.66 x 10(-14) m2 s( 1), respectively. Measurements of the effective membrane/concrete diffusion coefficient yielded a further average reduction in D of 98% for the Polyethylene Naphthalate and 96% for the PETG. Details of the experimental set-ups and procedures are described. The results of this investigation have shown that the application of an effective thin-film membrane adjacent to an intact concrete slab can significantly reduce the diffusion of radon gas entry. Therefore, the employment of a flexible thin-film membrane should be considered as a viable radon reduction technology method for residential new construction. PMID- 11379901 TI - Indoor radon concentrations caused by construction materials in 23 workplaces. AB - The aim of this study was to compare the measured and the calculated concentrations of indoor radon caused by building materials at 23 workplaces. The measured concentrations of radon were clearly higher than the calculated radon concentrations from the building materials, which indicated that the main source of indoor radon was the soil under and around the buildings. The highest means of continuously (933 Bq m(-3)) and integrated (169 Bq m(-3)) measured and calculated (from 70 to 169 Bq m(-3)) concentrations of radon were found in hillside locations. On the other hand, the median (27 and 43 Bq m(-3)) and maximum (626 and 1002 Bq m(-3)) values of calculated indoor radon concentrations exhaled from construction materials were the highest at the ground level places. On average, only 7-19% of the radon seemed to originate from the construction materials. PMID- 11379902 TI - Radon exhalation rate measurements on and around the premises of a former coal mine. PMID- 11379903 TI - Radiation performance index for Dutch dwellings: consequences for some typical situations. AB - This paper describes the new approach to control radiation exposure from natural sources to inhabitants of dwellings that is presently being considered in the Netherlands. The goal of this approach is to uphold the current rather favorable situation (average annual effective dose due to indoor radon and external radiation in dwellings is approx. 1 mSv). To achieve this goal a model is foreseen to predict the potential effective dose an inhabitant may receive from a dwelling on basis of its building plan. A scheme to calculate this dose is proposed in this paper. In future, such a scheme will be included in the Dutch Building Codes and houses to be built will be evaluated by using this scheme and comparing the results with, yet to be posed, limits to the potential effective dose. PMID- 11379904 TI - Results and conclusions of the Austrian radon mitigation project 'SARAH'. AB - The Austrian radon mitigation joint research project SARAH (supported by the Austrian Ministry of Economy and the Government of Upper Austria), a 2-year follow-up study of the Austrian National Radon Project (ONRAP), was started in 1996. The objectives of the research project were to find simple, cost-effective experimental methods for the characterisation of the radon situation in dwellings and to evaluate technically and economically the implementation of state of the art remedial actions for Austrian house types. After an intercomparison exercise of the assigned radon measuring instruments and detectors five houses were closely examined in regions with elevated radon levels in the federal state of Upper Austria. In this research work for the first time an extended Blower-Door method (which is conventionally used for determining the tightness of buildings) was successfully applied to radon diagnosis of buildings. In this paper the methods used for the radon diagnosis, the applied mitigation measures and the related technical and economical aspects are discussed. In conclusion of the results of this project a common strategy for solving the radon problem in Austria in the future is presented briefly. PMID- 11379905 TI - Experiences in radon-safe building in Finland. AB - A study was made of radon-safe buildings in 300 Finnish low-rise residential buildings using data obtained from a questionnaire study. The study also aims at finding the main defects in design and implementation and how the guidance given on radon-safe buildings in slab-on-grade houses has been followed. According to the guidelines, the prevention of the flow of radon-bearing air from the soil into the house is recommended to be carried out through installation of aluminised bitumen felt and use of elastic sealants. Second, as a precaution perforated piping should be installed in the subsoil of the floor slab. The median indoor radon concentration in the houses was 155 Bq/m3. This is 32% lower than the median of the estimated reference values. The action level of 200 Bq/m3 was still exceeded in 40% of the houses. In most houses with slab-on-grade the prevention was based only on the installation of a sub-slab depressurisation system. Sealing was performed in a low number of houses. In 80% of houses with a sub-slab piping connected to an operating fan, radon concentration was below the action level of 200 Bq/m3. In houses with piping but no fan, the corresponding fraction was only 45%. Sub-slab piping without a fan had no remarkable effect on radon concentration. In houses with crawl-space and edge-thickened slabs, radon concentrations were low. The choice of foundation system thus significantly affects the indoor radon concentration. The importance of complete and careful sealing work should be stressed in advice and guides concerning radon prevention. PMID- 11379906 TI - Radon risk management: the future challenge for the nuclear community. AB - The short-lived decay products (Rn-d) of radon gas (222Rn, 220Rn) have been identified as a health hazard in occupational exposure situations, as well as a public health risk in general. Decisions will have to be made concerning the initiation and scale of national Rn-mitigation programmes. In view of the potentially significant socio-economic impact of such programmes, it is recommended to proceed from the current approach based on Rn-d risk assessment to the integrated approach of radon risk management (RRM). A seven-step RRM is proposed, ranging from the selection of an action level based on an incremental risk-reduction, to the conduct of auditing procedures, monitoring the RRM outreach factor. PMID- 11379907 TI - Alpha particle emission from reference glass surfaces implanted with 210Po. AB - Implanted long-lived radon decay products in glass surfaces have been used as a measure of past radon exposure in homes. Special track-etch devices (so-called 'retro-detectors') attached to the glass surface, have the ability to specifically measure the implanted activity of 210Po in situ. Calibrating these devices for 210Po is fairly straightforward, but the retro-detectors are also sensitive to the background activity of the glass substrate. Thus, for the successful calibration of retro-detectors, it is necessary to determine the complete alpha emission energy spectrum of the reference glass sheet utilised as a calibration pad. In order to achieve accurate knowledge of the alpha surface emission rate, we have combined several different approaches, i.e. alpha spectrometry of the pad surface with both surface-barrier and pulse-ionisation detectors, and activity determination of the glass matrix by means of radiochemical methods. The part of the alpha emission spectrum originating from the glass volume is then calculated theoretically and compared with experimental results. PMID- 11379908 TI - Retrospective assessment of historic radon concentrations in Norwegian dwellings by measuring glass implanted Po-210--an international field intercomparison. AB - The first Norwegian study of historic radon concentrations in 17 dwellings in the high radon areas in Norway has been conducted as part of an international field intercomparison during 1998. The retrospective radon concentration is estimated via measurements of Po-210, the long-lived decay product of Rn-222 implanted in glass surfaces of objects like pictures, mirrors, cabinet-glass, etc. the method called 'surface trap'. Three different surface trap techniques to assess the implanted Po-210 activity and two different procedures to estimate retro radon from Po-210 data were used. The Po-210 and the retrospectively estimated radon results agree reasonably well over a wide range of concentrations. Historic radon concentrations were also estimated from analysis of a smaller number of 'volume trap' samples (pieces of spongy materials), and the results compared to those from 'surface traps'. The retro radon results correlate with contemporary radon results with a correlation coefficient of 0.877. To evaluate uncertainty in Po 210 measurements due to varying position on the glass a study of spatial homogeneity of three sample glasses was conducted and variations between 12% and 18% were found. PMID- 11379909 TI - Measurements of deposition velocity of radon decay products for examination of the correlation between air activity concentration of radon and the accumulated Po-210 surface activity. AB - The retrospective determination of radon exposure levels in dwellings by means of the measurement of the Po-210 surface activity is subject to various uncertainties. These result partly from the values assumed for the equilibrium factor F and for the unattached fraction f(p), and, more importantly, from differences in the deposition velocities of short-lived decay products of Rn-222, caused by varying conditions of turbulence. In order to evaluate the actual range of the variation which occurs under German living conditions, measurements for the deposition velocity parameter were carried out in several dwellings in which increased levels of radon were present. The statistical evaluation of the measurements produced a mean deposition velocity of 1.7 m/h for Po-218 and 0.4 m/h for Pb-214, and a relative standard deviation in the measured values of as low as approximately 50%. This lay significantly below the uncertainty value expected from the literature and would seem to justify the retrospective determination of the radon exposure from Po-210 surface activity measurement for use in, for example, epidemiological studies. PMID- 11379910 TI - Correlation of 210Po implanted in glass with radon gas exposure: sensitivity analysis of critical parameters using a Monte-Carlo approach. AB - In recent years, 210Po implanted in glass artefacts has been used as an indicator of the mean radon gas concentration in dwellings in the past. Glass artefacts have been selected in many dwellings and the alpha-recoil implanted 210Po concentration has been measured using various techniques. Some of these retrospective techniques use a model to estimate the retrospective radon gas on the basis of this surface 210Po activity. The accumulation of 210Po on glass surfaces is determined by the deposition regime over the exposure period. The 210Po activity is determined not only by the radon progeny deposition velocities, but by other room parameters such as ventilation rate, aerosol conditions and the surface to volume ratio of the room. Up to now in using room models, a nominal or 'base-case' scenario is used, i.e. a single value is chosen for each input parameter. In this paper a Monte-Carlo analysis is presented in which a probability distribution for each parameter is chosen, based on measurements quoted in the literature. A 210Po surface activity is calculated using a single value drawn from each of the parameter distributions using a pseudo-random number generator. This process is repeated n times (up to 20,000), producing n independent scenarios with corresponding 210Po values. This process permits a sensitivity analysis to be carried out to see the effect of changes in inputs on the model output. PMID- 11379911 TI - Radon risk mapping in southern Belgium: an application of geostatistical and GIS techniques. AB - A data set of long-term radon measurements in approximately 2200 houses in southern Belgium has been collected in an on-going national radon survey. The spatial variation of indoor Rn concentrations is modelled by variograms. A radon distribution map is produced using the log-normal kriging technique. A GIS is used to digitise, process and integrate a variety of data, including geological maps, Rn concentrations associated with house locations and an administrative map, etc. It also allows evaluation of the relationships between various spatial data sets with the goal of producing radon risk maps. Based on geostatistical mapping and spatial analysis, we define three categories of risk areas: high risk, medium risk and low risk area. The correlation between radon concentrations and geological features is proved in this study. High and medium Rn risk zones are dominantly situated in bedrock from the Cambrian to Lower Devonian, although a few medium risk zones are within the Jurassic. It is evident that high-risk zones are related to a strongly folded and fractured context. PMID- 11379912 TI - Approach to identification of radon areas in Germany. PMID- 11379913 TI - Mapping the geogenic radon potential in Germany. AB - Mapping the geogenic radon potential in Germany is a research project initiated by the German Federal Ministry for the Environment, Conservation and Reactor Safety. The project was aimed to develop a standard methodology for the estimation of a geogenic radon potential and to apply this method to map the region of Germany as an overview for planning purposes. The regionalisation results from a distance-weighted interpolation of the site-specific values of radon concentration in soil gas and in situ gas permeability of soils on a regular grid considering the corresponding geological units. The map of Germany in a scale of 1:2 million is based on the radon concentration in soil gas as an estimator of the geogenic radon potential assuming the 'worst case' of uniform highest permeability. The distribution is subdivided into categories of low (< 10 kBq/m3), medium (10-100 kBq/m3), increased (100-500 kBq/m3) and high (> 500 kBq/m3) radon concentration. High values occur especially in regions with granites and basement rocks of Paleozoic age, and are proven by measurements in 0.03% of the total area. Many of these regions are also known for their enhanced indoor values. The class with increased values takes a portion of 7.86% and likewise occurs mainly in regions with outcrops of folded and metamorphic basement, but also of some Meso- and Cenozoic sediments with increased uranium contents and/or higher emanation coefficients. For 67.3% of the country, the radon concentration is classified as 'medium', and an assignment to specific geological units cannot be made at the map scale considered. Low radon contents, where protective measures against radon are usually not considered, are found in the geologically rather homogeneous part of northern Germany with unconsolidated Cenozoic sediments, covering approximately 25% of the total country. It is of course not possible to predict the indoor radon concentration of single houses from these maps, because construction type and structural fabric of houses are essentially governing the extent to which subsoil radon potential affects the indoor concentration. Besides this, in places with site-specific geochemical, structural and soil-physical properties, local radon anomalies may occur which were not recorded in the course of the wide-meshed screening study. PMID- 11379914 TI - The cost effectiveness of radon remediation programmes in hospitals, schools and homes in radon affected areas in the UK. PMID- 11379915 TI - Mapping indoor radon-222 in Denmark: design and test of the statistical model used in the second nationwide survey. AB - In Denmark, a new survey of indoor radon-222 has been carried out, 1-year alpha track measurements (CR-39) have been made in 3019 single-family houses. There are from 3 to 23 house measurements in each of the 275 municipalities. Within each municipality, houses have been selected randomly. One important outcome of the survey is the prediction of the fraction of houses in each municipality with an annual average radon concentration above 200 Bq m(-3). To obtain the most accurate estimate and to assess the associated uncertainties, a statistical model has been developed. The purpose of this paper is to describe the design of this model, and to report results of model tests. The model is based on a transformation of the data to normality and on analytical (conditionally) unbiased estimators of the quantities of interest. Bayesian statistics are used to minimize the effect of small sample size. In each municipality, the correction is dependent on the fraction of area where sand and gravel is a dominating surface geology. The uncertainty analysis is done with a Monte-Carlo technique. It is demonstrated that the weighted sum of all municipality model estimates of fractions above 200 Bq m(-3) (3.9% with 95%-confidence interval = [3.4,4.5]) is consistent with the weighted sum of the observations for Denmark taken as a whole (4.6% with 95%-confidence interval = [3.8,5.6]). The total number of single family houses within each municipality is used as weight. Model estimates are also found to be consistent with observations at the level of individual counties. These typically include a few hundred house measurements. These tests indicate that the model is well suited for its purpose. PMID- 11379916 TI - The outcome of the Lithuanian radon survey. PMID- 11379917 TI - The present status of the radon program in the United States of America. PMID- 11379918 TI - Thoron spike test for passive radon detectors. PMID- 11379919 TI - Uncertainty, variability and sensitivity analysis applied to the RAGENA model of radon generation, entry and accumulation indoors. AB - The application of a radon model is useful to understand the processes that drive the radon gas behaviour from its sources to its accumulation indoors. Since in a given inhabited house the detailed knowledge of the values of all the parameters that affect indoor radon levels is not available, the response of the model has to be explored in a reference site in which all the parameters are supposed to be known. We call this site the reference configuration. In this paper we report on the procedure followed to carry out uncertainty, sensitivity and variability analysis of the model response for a reference configuration that corresponds to a single family multi-zone house. We have obtained from the uncertainty analysis that, assuming a normal distribution of all the input parameters with a 10% relative standard deviation (R.S.D.), the model outputs present a R.S.D. in the range 17-22%. The sensitivity analysis reflects, in general, a good behaviour of the model, in the sense that its response describes a realistic behaviour of the system. The variability analysis has shown that the model is applicable to a wide range of situations, and that the most relevant parameters for the reference configuration are: the soil gas-permeability (obtained from the mean soil grain diameter); the ventilation rate of the rooms; the air-exchange rate between the basement and room 2; the soil-indoor pressure difference; the open area; and the concrete radium content. PMID- 11379920 TI - Integrated natural radiation exposure studies in stable Yugoslav rural communities. AB - The results of field investigations of natural radiation exposures of the general population in two stable rural communities in Yugoslavia are presented. The principal emphasis was on exposures to contemporary indoor radon, but measurements of external penetrating radiation absorbed dose rates in air were carried out in the majority of cases. In addition, in a limited number of dwellings, measurements of thoron gas concentrations were made. By means of making a series of sequential 3-month radon measurements, both seasonal variations and annual average radon levels in the dwellings were determined. Using passive alpha track detectors, individual radon and thoron indoor concentrations as high as 9591 Bq m(-3) and 709 Bq m(-3), respectively, were detected while absorbed dose rates in air in the dwellings as high as 430 nGy h( 1) were recorded. On the basis of these different types of measurements, assessments could be made of the integrated natural radiation exposures being received by the populations. In addition to contemporary radon measurements, retrospective radon exposure assessments in most of the dwellings were made on the basis of measurements of 210Po concentrations in both surface (glass) traps and in volume (porous materials) traps. A description is given of the sampling strategies and protocols used in this field work. It is shown that at least one stable rural community receiving high natural radiation exposures, has been clearly identified and plans for future health investigations of the population there are outlined. PMID- 11379921 TI - Radioenvironmental survey of the Megalopolis lignite field basin. AB - The Megalopolis lignite field basin in southern Greece, with Megalopolis-A and B lignite-fired power plants in operation (total 900 MW), has been repeatedly investigated during the past 25 years by the Nuclear Engineering Section of the National Technical University of Athens (NES-NTUA). The present work aims at an integrated radioenvironmental approach leading to the dose assessment to the public and to the plants staff. This approach includes systematic sampling of lignite and barren at the local lignite mines feeding the power plants and sampling of lignite, fly-ash and bottom ash at the power plants for the determination of the activity of the natural radionuclides 226Ra, 232Th, 40K, 234Th and 210Pb. Furthermore, the following measurements and samplings were conducted in 25 selected sites within 10 km around the power plants: soil sampling for the determination of the above radionuclides, radon concentration and exhalation rate measurements, soil gas radon concentration measurements, dose measurements and calculations, determination of air-particulate matter concentration, etc. The results obtained allowed for the mapping of the parameters studied which lead to useful conclusions. Dosimetric calculations for the population living around the power plants and the plants staff were also performed based on the guidance of UNSCEAR (1982 report). PMID- 11379922 TI - Radon concentration in soil gas: a comparison of the variability resulting from different methods, spatial heterogeneity and seasonal fluctuations. AB - From the end of 1996 through March 1999, the spatial and temporal variability of the soil 222Rn concentration was investigated at a 20 m x 20 m test field with porous soil in 0.5 m and 1.0 m depth at nine positions each and at 1 m x 1 m plots at four positions each. For this, soil gas was collected weekly into evacuated scintillation cells and was analysed subsequently for radon activity. In the 20 m x 20 m field the spatial variability was characterized by coefficients of variation (C.V.) of 26% at 0.5 m, and 13% at 1.0 m depth. Within the 1 m x 1 m plots the C.V. values were 4% and 2%, i.e. within the uncertainty of the method. Time series analysis (TSA) of the soil radon data shows seasonal variations with maximum concentrations in the winter months. Radon concentrations ranged from 6 to 50 kBq m(-3) in 0.5 m depth, and from 8 to 34 kBq m(-3) in 1.0 m depth. Mostly, the concentrations were higher in 0.5 depth than in 1.0 m depth. However, seasonal variation of the 0.5 m to the 1.0 m concentration ratio has been verified by TSA. To test the variability resulting from different methods, additional procedures and instruments were investigated at the 20 m x 20 m field and at a second test field with a different soil type. Soil gas sampling into evacuated scintillation cells was selected as the reference procedure. Soil radon concentrations obtained with the different sampling procedures and detection methods at the 20 m x 20 m field essentially agreed within the limits of uncertainty of the methods tested. At the second test field, i.e. in a largely impermeable soil, deviations up to a factor of two related to the reference procedure were observed. PMID- 11379923 TI - Correlations between radon concentration and indoor gamma dose rate, soil permeability and dwelling substructure and ventilation. AB - Correlations between radon concentration and indoor gamma dose rate, soil permeability and dwelling substructure and ventilation were studied using data from 84 low rise residential houses collected in an area of enhanced indoor radon concentration. The radon concentrations varied from 30 to > 5000 Bq m(-3). Cross tabulation, comparisons of means and multiplicative models were used to test the significance of the effects. In this study a quite high percentage of explained variation R2 (68%) was found. It was found that the most important factors were the substructure and the permeability of the soil. Due to the rather small sample size and moderate variation in the uranium content of the bedrock of the area, the effect of the indoor gamma dose rate was not so prominent. The effects of ventilation habits and sleeping with open windows were not detected in this study. PMID- 11379924 TI - Field experience with volume traps for assessing retrospective radon exposures. AB - Approximately 200 volume traps were retrieved from dwellings in various radon prone areas in Europe. They were analysed for the purpose of retrospective radon assessment. Emphasis is put on specific problems encountered when using field samples as opposed to laboratory exposed samples. It was seen that in very dusty circumstances, direct penetration of radon decay products from the outside to the centre of the volume traps calls for extra caution. Rinsing the samples is proposed as a solution and was tested in field and laboratory conditions, showing good results. An attempt was made to give an assessment of the achievable accuracy of the method. Where possible, the volume trap retrospective results were compared with contemporary measurements or to retrospective results from surface traps. The overall impression is that although volume traps are sometimes hard to find in the field, the high reliability of the results makes it well worth the effort. PMID- 11379925 TI - Challenges in harmonising controls on the radioactivity of building materials within the European Union. AB - Possibilities for harmonising controls on the radioactivity of building materials within the European Union are being discussed in a Working Party on Natural Radiation Sources established by the Article 31 Group of Experts (Euratom Treaty). The Working Party is preparing a document to aid the Article 31 Expert Group and the European Commission in considering possible recommendations and technical guidance to the Member States for the implementation of the new Basic Safety Standards Directive concerning the radioactivity of building materials. The discussions in the working party have demonstrated several challenges in the possible harmonisation of controls, many of them arising from some significant differences in national circumstances. PMID- 11379926 TI - Field tests of a radon progeny sampler for the determination of effective dose. AB - A two-screen sampler (an effective dosimeter), with a collection efficiency matched to the particle size response of the radon progeny dose conversion factors (DCF), obtained from the ICRP respiratory tract model as implemented in the computer code RADEP, has been developed to assess the inhalation dose from exposure to radon progeny. In order to evaluate the performance of this sampler, the second stage of a six-stage wire screen diffusion battery was designed to operate as an Effective Dosimeter. This hybrid system allowed two methods for the determination of the radon progeny DCF. For the first method, the activity size distributions, measured using the diffusion battery, were used to obtain a size weighted DCF. A second determination of DCF was obtained directly from the fraction collected by the Effective Dosimeter. The hybrid diffusion battery was used to measure radon progeny in the Fairy Cave, Buchan, Victoria at 20-min intervals over a 30-h period. This cave had radon concentrations exceeding 2000 Bq m(-3), with low aerosol concentration and low ventilation rates. The measurements were analysed for the radon progeny PAEC, the activity size distribution, the size-weighted DCF and the effective dosimeter collected fraction. The Effective Dosimeter DCFs were determined from the collected fraction using firstly a simple linear function and then using a more complex polynomial function to correct for residual errors. For the linear factor alone, the calculated Effective Dosimeter DCFs were on average 11% lower than the equivalent size-weighted DCF values. The agreement using the polynomial function was improved markedly, with a linear regression of the DCF yielding a fitted ratio of 0.965, with an R value of 0.99. For this study, the use of the ICRP conversion convention to estimate the occupational exposure to the tour guides working in the Fairy Cave would under estimate the effective dose by up to a factor of 2. PMID- 11379927 TI - Experimental set-up for measuring diffusive and advective transport of radon through building materials. AB - This study describes an approach for measuring and modelling diffusive and advective transport of radon through building materials. The goal of these measurements and model calculations is to improve our understanding concerning the factors influencing the transport of radon through building materials. To reach this goal, a number of experiments have to be conducted. These experiments, including measurements in a large cylinder for creating diffusive and advective transport of radon under controlled, 'dwelling-like' conditions, are described here and the initial results are presented. A better understanding about the transport of radon through building materials will lead to more effective ways to decrease or to prevent the entrance of radon into dwellings. PMID- 11379928 TI - Methods for measuring diffusion coefficients of radon in building materials. AB - Two methods for determining the Rn-222 diffusion coefficient in concrete are presented. Experimentally, the flush and adsorption technique to measure radon release rates underlines both methods. Theoretically, the first method was developed for samples of cubical geometry. The radon diffusion equation was solved for boundary conditions imposing zero flux conditions successively on each side. In practice, a 100% effective covering would ensure this condition to be satisfied. The diffusion coefficient is then determined by comparing the computed and respectively measured normalized ratios of the radon release rates with respect to the rate corresponding to the open-boundaries (uncovered) specimen. As in practice, none of the investigated coverings showed to be effective in reducing radon exhalation, indicating that radon-tight sealing of surfaces is far from trivial, no clear conclusions could be drawn with respect to the diffusion coefficient. The second method can be applied to specimens that are first reshaped into hollow cylinders. A one-dimensional situation can be inforced by requiring that the flux vanishes at the two ends of the hollow cylinder. The theoretical and experimental ratios are again compared. In practice, the radon flux originates from a radon source enclosed inside the hollow cylinder, the effectiveness of the sealing being previously tested on an aluminum dummy of similar dimensions. The radon bulk diffusion coefficient for the used concrete sample resulted in a value of D = (4.6 +/- 0.4) x 10(-6) cm2 s(-1). PMID- 11379929 TI - Numerical modelling of radon-222 entry into houses: an outline of techniques and results. AB - Numerical modelling is a powerful tool for studies of soil gas and radon-222 entry into houses. It is the purpose of this paper to review some main techniques and results. In the past, modelling has focused on Darcy flow of soil gas (driven by indoor-outdoor pressure differences) and combined diffusive and advective transport of radon. Models of different complexity have been used. The simpler ones are finite-difference models with one or two spatial dimensions. The more complex models allow for full three-dimensional and time dependency. Advanced features include: soil heterogeneity, anisotropy, fractures, moisture, non uniform soil temperature, non-Darcy flow of gas, and flow caused by changes in the atmospheric pressure. Numerical models can be used to estimate the importance of specific factors for radon entry. Models are also helpful when results obtained in special laboratory or test structure experiments need to be extrapolated to more general situations (e.g. to real houses or even to other soil-gas pollutants). Finally, models provide a cost-effective test bench for improved designs of radon prevention systems. The paper includes a summary of transport equations and boundary conditions. As an illustrative example, radon entry is calculated for a standard slab-on-grade house. PMID- 11379930 TI - Soil gas measurements below foundation depth improve indoor radon prediction. AB - A soil gas measurement method developed earlier, [Nucl Tracks Radiat Meas, 22(1 4) (1993) 468] was applied to boreholes drilled to below foundation depth. Radon concentration and permeability were measured at 50-cm intervals. In radon prone areas, permeability showed an increase with depth over several orders of magnitude, indicating a low permeability top layer with a thickness of 0.5 m and more. A radon availability index (RAI) was empirically defined and the maximum RAI of each boring proved to be a reliable indicator for radon problems in nearby houses. The permeability of the top layer also proved to be an important factor for a better understanding of soil gas transport and the influence of rain. Implications for radon mitigation are derived. PMID- 11379931 TI - Radon exposure vs. exposure to other sources of ionising radiation. PMID- 11379932 TI - The reliability of radon reduction techniques. PMID- 11379933 TI - In situ gamma spectroscopy to characterize building materials as radon and thoron sources. AB - In situ gamma spectroscopy is widely utilized to determine the outdoor gamma dose rate from the soil and to calculate the natural and artificial radionuclide concentration and their contribution to the dose rate. The application of in situ gamma spectroscopy in indoor environments can not supply quantitative information about activity concentration of radionuclides in building materials, but this technique can provide interesting information about building materials as radon source. In fact, a method based on analyses of gamma spectra data has been developed by the authors to provide, in field, quantitative estimation of disequilibrium in 226Ra and 228Ac sub-chains due to 222Rn and 220Rn exhalation. The method has been applied to data of gamma spectroscopy measurements carried out with HPGe detector (26%) in seven dwellings and one office in Rome. The first results of the data analysis show that, as regards especially the 226Ra sub-chain disequilibrium, different building materials (tuff, concrete, etc.) can show very different characteristics. If, in addition to the spectrometric data, other indoor environment parameters (indoor gamma dose rates, room dimensions, wall thickness, etc.) (Bochicchio et al., Radiat Prot Dosim 1994;56(1-4):137-140; Bochicchio et al., Environ Int 1996a;22:S633-S639) are utilized in a room model, an evaluation of 226Ra, 228Ac and 40K activity concentration and an indication of the exhalation features, by means of estimation of exhaled 222Rn activity concentration, can be achieved. PMID- 11379934 TI - Results of a simple intercomparison of natural radioactivity measurements using a 'blue concrete' sample. AB - A simple intercomparison of natural radioactivity and radon emanation factor measurements co-ordinated by the Medical Physics Department of the University of Cantabria UC, Spain, has been carried out during 1998 in the framework of the EU Concerted Action ERRICCA (European Research into Radon in Construction Concerted Action). All the measurements have been made on a 'blue concrete' sample kindly donated by Dr G. Akerblom from the Swedish Radiation Protection Institute. In addition to UC, two other participant laboratories, STUK from Finland and ZVD from the Republic of Slovenia, have contributed to the development of the intercomparison exercise. Considering the uncertainties of the measurements, a good agreement between the results obtained by the three participating laboratories has been ascertained. However, in accordance with experimental results obtained by other authors, data reported by UC show a significant decrease of the radon emanation factor as moisture content of the sample decreases below 5% (per weight). PMID- 11379935 TI - Teaching radon in the living environment: school experiments with air, liquids and solids. PMID- 11379936 TI - Radon exposure and lung cancer risk--Czech cohort study on residential radon. AB - Epidemiological evidence of lung cancer risk from radon is based mainly on studies of men employed underground in mines where exposures are relatively high in comparison to indoor exposure. Nevertheless, direct evidence of risk from residential radon is desirable. In 1990, a study was started comprising 12,000 inhabitants of an area with elevated radon concentrations. The mean level in the houses was higher than general mean of the country by a factor of five. In the period 1961-1995, a total of 173 lung cancers were observed. Comparing to nationally expected numbers (E), the observed number (O) of cases is elevated (O/E = 1.11), in contrast to generally low figures for cancers other than lung (O/E = 0.85). Lung cancer risk related to cumulative exposures experienced in the past 5-24 or 5-35 years were both significant. In relation to standard radon progeny concentration 100 Bq/m3, the excess relative risk coefficient was 0.103 (95% CI, 0.039-0.168), the value somewhat lower than findings in other indoor studies. PMID- 11379937 TI - Approaches to the assessment of long term exposure to radon and its progeny. AB - In recent years, a number of case-control epidemiological studies have taken place and others are in progress to evaluate the lung cancer risk to the general population from exposure to radon and its short-lived progeny in the indoor residential environment. While it is actually long term exposure over past decades to radon progeny by inhalation that dominates lung doses, for a number of practical reasons it is radon gas that is measured in these studies. Because the risk from radon and its progeny results from cumulative exposure over past decades rather than from contemporary exposure, it is necessary to reconstruct the historical exposures of subjects. A number of factors limit the accuracy of this approach of which the following are perhaps the most important: the mobility and residential history of the subjects; radon exposures elsewhere; and changes that may have occurred in the radon levels in current and previous residences. Measurement techniques to assist in making more direct retrospective assessments of radon exposure have appeared in the recent past and are the subject of this paper. These are based on the measurement of the long-lived radon progeny 210Po trapped in household artefacts such as glass or porous and spongy materials. In vivo measurements of skeletal 210Pb in exposed persons is also a method that is currently being investigated as a means to assess historical exposures to radon. The advantages and disadvantages of these methods are described here as well as their potential in future radon epidemiological studies. PMID- 11379938 TI - Experience from retrospective radon exposure estimations for individuals in a radon epidemiological study using solid-state nuclear track detectors. AB - The relation between increased risk of lung cancer and exposure to indoor radon is assessed in epidemiological studies. Both the quality and reliability of smoking data and the radon exposure data are of primary importance. Contemporary measurement of radon concentration in the dwellings of individuals in a case control study is traditionally used to assess past history of radon exposure. These assessments are somewhat unreliable since presently measured radon concentration might not be representative for a given location long ago. The measurement of long-lived decay products from 222Rn remaining indoors on hard surfaces, such as glass, makes it possible to assess the exposure to indoor radon. At the Swedish Radiation Protection Institute, a combination of two different solid-state nuclear track detectors has been developed to assess the 210Pb activity implanted in glass surfaces by measuring 210Po alpha activity. This detector (a RETRO detector) is used in the Swedish radon epidemiological case-control study of non-smokers with the aim to provide an alternative estimate of individual radon exposure and to evaluate the usefulness of RETRO measurements. A total of 576 different objects were found and 568 were measured. For 225 individuals, we measured two personal objects that had been in the same person's possession for more than 20 years. The standard deviation of the average radon concentration obtained from these two objects had a median value of 13 Bq/m3 indicating a precision of exposure of approximately 20%. The correlation between 210Po surface activity measured earlier and the mean values of radon concentrations in a number of Swedish dwellings is used to estimate the historical, average radon concentration. This average correlation factor seems also to be valid for measurements in the non-smoker epidemiological study. PMID- 11379939 TI - The Iowa radon lung cancer study--phase I: Residential radon gas exposure and lung cancer. AB - Exposure to high concentrations of radon (222Rn) progeny produces lung cancer in both underground miners and experimentally-exposed laboratory animals. The goal of the study was to determine whether or not residential radon exposure exhibits a statistically significant association with lung cancer in a state with high residential radon concentrations. A population-based, case-control epidemiologic study was conducted examining the relationship between residential radon gas exposure and lung cancer in Iowa females who occupied their current home for at least 20 years. The study included 413 incident lung cancer cases and 614 age frequency-matched controls. Participant information was obtained by a mailed-out questionnaire with face-to-face follow-up. Radon dosimetry assessment consisted of five components: (1) on-site residential assessment survey; (2) on-site radon measurements; (3) regional outdoor radon measurements; (4) assessment of subjects' exposure when in another building; and (5) linkage of historic subject mobility with residential, outdoor, and other building radon concentrations. Histologic review was performed for 96% of the cases. Approximately 60% of the basement radon concentrations and 30% of the first floor radon concentrations of study participants' homes exceeded the US Environmental Protection Agency action level of 150 Bq m(-3) (4 pCi l(-1)). Large areas of western Iowa had outdoor radon concentrations comparable to the national average indoor value of 55 Bq m( 3) (1.5 pCi l(-1)). Excess odds of 0.24 (95% CI = -0.05-0.92) and 0.49 (95% CI = 0.03-1.84) per 11 WLM(5-19) were calculated using the continuous radon exposure estimates for all cases and live cases, respectively. Slightly higher excess odds of 0.50 (95% CI = 0.004-1.80) and 0.83 (CI = 0.11-3.34) per 11 WLM(5-19) were noted for the categorical radon exposure estimates for all cases and the live cases. A positive association between cumulative radon gas exposure and lung cancer was demonstrated using both categorical and continuous analyses. The risk estimates obtained in this study indicate that cumulative radon exposure presents an important environmental health hazard. PMID- 11379940 TI - Ventilation and radon transport in Dutch dwellings: computer modelling and field measurements. AB - In 1995 and 1996 radon concentrations and effective air flows were measured in approximately 1500 Dutch dwellings built between 1985 and 1993. The goal of this investigation was to describe the trend in the average radon concentration by supplementing the first survey on dwellings built up to 1984 and to quantify the contributions of the most important sources of radon. In the living room of new dwellings the average radon concentration was 28 Bq m(-3), which is 50% higher than in dwellings built before 1970. Measurements of effective air flows showed the most important source of radon in the living room of new dwellings to be the building materials, with an average contribution of 70%. The other 30% comprised outside air and air from the crawl space in equal quantities. The long-term increase in the indoor radon concentration is mainly due to improvements in insulation since 1970, resulting in a fourfold decrease in infiltration through the building shell. Model calculations, supplementing the field measurements, confirmed the dominant effect of increasing airtightness of dwellings compared to effects of the observed trend in the use of building materials. PMID- 11379941 TI - Applicability of various insulating materials for radon barriers. AB - The effectiveness of various insulating materials for limiting radon entry into houses has been investigated under laboratory conditions. Results for the radon diffusion coefficient measurements in more than 80 insulating materials are summarized. We have discovered that great differences exist in diffusion properties, because the diffusion coefficient varies within four orders from 10( 13) m2/s to 10(-10) m2/s. A methodological approach is proposed in order to identify the minimal thickness of radon-proof membranes, depending on building and soil characteristics. General guidelines for the selection of radon-proof insulation are presented. PMID- 11379942 TI - Radon permeability and radon exhalation of building materials. AB - High radon concentrations indoors usually depend on the possibilities of radon penetration from the surrounding soil into the buildings. Radon concentrations in dwellings up to 100 kBq/m3 were found in some special regions (i.e. Schneeberg/Saxony, Umhausen/Tyrol), where the soil shows a high uranium content and additionally, a fast radon transport in the soil is possible. To reduce the radon exposure of the inhabitants in these 'radon prone areas' it is necessary to look for building and insulating materials with low radon permeability. We examined several building materials, like cements, concretes and bricks of different constitutions for their diffusion coefficients and their exhalation rates. The insulating materials, like foils and bitumen were tested also on their radon tightness. The measurements were performed with an online radon measuring device, using electrostatic deposition of 218Po ions onto a surface barrier detector and subsequent alpha spectroscopy. The mean diffusion lengths for the investigated building materials range from lower than 0.7 mm (i.e. for plastic foil), up to 1.1 m for gypsum. The diffusion length R was calculated from the diffusion coefficient D with R = square root(D/lambda). If the thickness of the material is more than 3 times the diffusion length, then it is called radon tight. The mean 222Rn exhalation rates for the building materials varied between 0.05 and 0.4 mBq/m2s. The samples were investigated as stones, plates, blocks, foils, coatings, powders etc., no statement can be made about working at the construction site of a building. Also the fabrication and processing of the materials has to be considered, because the material characteristics may have changed. PMID- 11379943 TI - Cost effectiveness analysis of radon remediation programmes. AB - The economic implications of regulations governing radon gas level identification and remediation in buildings are poorly understood, and attempts to address these issues have been criticised for lack of comparability. It is imperative therefore that a general model for the economic evaluation of radon remediation programmes is adopted to ensure comparability between studies and settings and to increase the usefulness of the results to decision makers. This paper presents general guidelines for the use of cost-effectiveness analysis (CEA) as an economic appraisal tool in the evaluation of radon reduction and prevention programmes. The data requirements for a CEA of radon remediation programmes concern both costs and outcomes. These components are discussed with respect to: programme objectives, comparator choice, perspective, time horizon, discounting, uncertainty, and final ratios. Adhering to clear guidelines concerning these aspects of evaluations will facilitate meaningful evaluation of radon remediation programmes. Finally, by evaluating the radon remediation programmes using methods applied to other health interventions (such as lung cancer prevention interventions), comparisons using the same metric can be made across policy areas. PMID- 11379944 TI - Radon protection for new buildings: a practical solution from the UK. AB - If indoor radon levels are to be significantly reduced across Europe it is essential to ensure that all new buildings built in areas affected by radon are protected. In the United Kingdom the Building Research Establishment Ltd (BRE) has been carrying out research on behalf of the Department of the Environment, Transport and the Regions (DETR) to develop protective measures for use in new buildings. This work commenced in the mid-1980s and has resulted in the development of a range of practical cost-effective techniques for providing radon protection in new UK buildings. Guidance has been developed in support of the Building Regulations for England and Wales. First published in 1991 the technical solutions have been gradually improved in light of experience gained on site. Likewise, the areas for which the guidance applies have also been revised in light of radon surveys carried out by the National Radiological Protection Board. This continuous process of refinement is scheduled to result in a further version during 1999. The techniques developed in the UK mainly rely upon passive radon barriers, which are low cost and simple to install. Even in the worst affected areas of the country, where some 30% of existing houses have radon levels greater than 200 Bq/m3 (UK recommended action level), houses with protection measures regularly result in indoor radon levels close to 20 Bq/m3 (UK average for all house types). Radon protective measures are now being installed routinely in many parts of the UK. The cost of installation has reduced as protection has become a matter of routine through regulation, so that currently radon protection is unlikely to cost much more than 0.25% of the total construction cost of a typical UK house. It is our view that many of the techniques developed in the UK could be used directly or with minor modifications in other countries in Europe. This paper describes these latest techniques and approaches used in the UK to successfully provide radon protective measures. PMID- 11379945 TI - Size distribution, equilibrium ratio and unattached fraction of radon decay products under typical indoor domestic conditions. AB - In order to characterise the behaviour of radon decay products under domestic conditions, long-term measurements were carried out from May 1997 to April 1998 in a typical dwelling located in Brittany (France). In particular, the unattached fraction and equilibrium factor were continuously measured. Moreover, the size distributions of unattached and attached radon daughters were investigated by using specific instruments implemented in the laboratory. All these experiments were carried out under different typical aerosol conditions. The results evidenced the strong influence exerted by the characteristics (concentration, size) of ambient aerosol on these different parameters. PMID- 11379946 TI - Recent advances in capillary electrophoresis/electrospray-mass spectrometry. AB - In this review, the progress in hyphenation of capillary electrophoresis (CE) with electrospray ionization-mass spectrometry (ESI-MS) since the article of Banks (Banks, J. F., Electrophoresis 1997, 18, 2255-2266) is reported. In all capillary-based electromigration techniques, such as capillary gel electrophoresis (CGE), capillary isotachophoresis (CITP), capillary isoelectric focussing (CIEF), micellar electrokinetic chromatography (MEKC), affinity capillary electrophoresis (ACE), as well as in the hybrid techniques capillary electrochromatography (CEC), and pressurized capillary electrochromatography (pCEC) progress has been made in experimental setups, and for many groups of analytes, such as peptides, proteins, nucleotides, saccharides, drugs and their metabolites, CE/ESI-MS has been successfully applied. Electromigration is further miniaturized. New preconcentration methods allow the investigation of compounds, which are not sensitively detected with ESI-MS. Coordination ion spray (CIS) MS is another method for sensitivity enhancement by on-line formation of charged coordination compounds. PMID- 11379947 TI - Estimation of the dissociation constants for functional groups on modified and unmodified silica gel supports from the relationship between electroosmotic flow velocity and pH. AB - Electroosmotic volume flow was directly observed in a simple instrument consisting of 1 cm long a home-made support, packed between two polyethylene frits in the polypropylene tube. Equations relating electroosmotic flow (EOF) velocity and pH for two functional groups on the surface of the solid materials were developed. With these equations, we can estimate the dissociation constants of two different kinds of functional groups on modified silica gel materials simultaneously. The dissociation constants of silanol groups, benzene sulfonic acid groups, and alkyl quaternary ammonium groups on the modified and unmodified silica gel supports were estimated. The estimated pK values of the silanol groups on the silica gel and modified silica gel surfaces are between 4.0 and 4.3. The estimated pK values of the benzene sulfonic acid groups and alkyl quaternary ammonium groups on the surface of the modified silica gel are 2.6 and 8.6, respectively. PMID- 11379948 TI - Anion-exchange capillary electrochromatography with indirect UV and direct contactless conductivity detection. AB - Conductivity detection is applied to ion-exchange capillary electrochromatography (IE-CEC) with a packed stationary phase, using a capacitively coupled contactless conductivity detector with detection occurring through the packed bed. Columns were packed with a polymeric latex-agglomerate anion-exchanger (Dionex AS9-SC). A systematic approach was used to determine suitable eluants for IE-CEC separations using simultaneous indirect UV and direct conductivity detection. Salicylate and p-toluenesulfonate were identified as potential eluant competing anions having sufficient eluotropic strength to induce changes in separation selectivity, but salicylate was found to be unsuitable with regard to baseline stability. It was also found for both indirect UV and direct conductivity detection that homogenous column packing was imperative, and monitoring of the baseline could be used to assess the homogeneity of the packed bed. Using a p-toluenesulfonate eluant, the separation of eight common anions was achieved in 2.5 min. Direct conductivity detection was found to be superior to indirect UV detection with regard to both baseline stability and detection sensitivity with detection limits of 4-25 microg/L being obtained. However, the calibration for each anion was not linear over more than one order of magnitude. When using conductivity detection, the concentration of the eluant could be varied over a wider range (2.5-50 mM p toluenesulfonate) than was the case with indirect UV detection (2.5-10 mM), thereby allowing greater changes in separation selectivity to be achieved. By varying the concentration of p-toluenesulfonate in the eluant, the separation selectivity could be manipulated from being predominantly ion-exchange in nature (2.5 mM) to predominantly electrophoretic in nature (50 mM). PMID- 11379949 TI - Enantioseparations using nonaqueous capillary electrochromatography on cellulose and amylose tris(3,5-dimethylphenylcarbamates) coated on silica gels of various pore and particle size. AB - Enantioseparations of chiral compounds were studied in nonaqueous capillary electrochromatography (NAQ CEC) with cellulose and amylose tris(3,5 dimethylphenylcarbamates) (Chiralcel OD and Chiralpak AD, respectively) coated on the silica gels of various pore and particle size. Increasing intraparticle perfusive transport with increasing pore size of silica favorably affected peak efficiency and resolution of enantiomers, although some decrease of separation factor was observed in the pore size range 60-200 A. Further improvement of peak efficiency was observed when the particle size of silica was reduced from 5 to 3 microm. The effects of a separation medium and temperature are also reported and the data obtained in the same capillaries in CEC and capillary liquid chromatography (LC) mode are compared. PMID- 11379950 TI - Evaluation of ODS-AQ stationary phase for use in capillary electrochromatography. AB - The aim of this study was to evaluate the applicability of ODS-AQ packing material as a stationary phase in capillary electrochromatography (CEC). The electroosmotic flow created on an ODS-AQ stationary phase was measured at different mobile phase compositions and at different column temperatures. It was observed that the electroosmotic flow generated in the column increased by 50% when the temperature of the system was raised from 20 degrees C to 60 degrees C, while all other conditions were kept constant. The electroosmotic flow produced by the ODS-AQ stationary phase was found to be comparable to the flow generated in a column packed with Nucleosil bare-silica material. In addition, a set of polar compounds (D-lysergic acid diethylamide derivatives) was utilized to determine the influence of temperature and mobile phase composition on their chromatographic behavior on an ODS-AQ stationary phase in a CEC mode. A linear relationship between the solute retention factor and column temperatures was seen over the temperature range studied (20 degrees C to 60 degrees C). A quadratic function was used to describe the changes in the solute retention factors with variation of acetonitrile concentration in the mobile phase. PMID- 11379951 TI - Separation of 4-dimethylamino-6-(4-methoxy-1-naphthyl)-1,3,5-triazine-2-hydrazine derivatives of carbonyl compounds by reversed-phase capillary electrochromatography. AB - 4-Dimethylamino-6-(4-methoxy-1-naphthyl)-1,3,5-triazine-2-hydrazine (DMNTH) is a novel derivatizing reagent specially designed for the determination of carbonyl compounds. In this work, we describe the separation of DMNTH-derivatized carbonyl compounds by reversed-phase capillary electrochromatography (CEC). After systematic investigations of the effects of experimental conditions viz. pH and concentration of buffer, type of stationary phase, injection volume of sample, organic modifier, and temperature, optimal conditions were found. The sample compounds, which were separated with gradient high performance liquid chromatography (HPLC), were separated by CEC under isocratic elution due to the high efficiency. Comparisons of separations by CEC and micellar electrokinetic chromatography (MEKC) were made. PMID- 11379952 TI - Study on liposomes by capillary electrophoresis. AB - Liposomes made of mixtures of zwitterionic and anionic lipids were investigated by means of capillary electrophoresis and dynamic light scattering. The influence of the molar lipid ratio and of the buffers, used in the running electrolyte solution, on the physical characteristics of the liposomes were investigated. Data on effective electrophoretic mobilities, total charges as well as sizes of the liposomes are given. In addition, examples on the use of liposomes as carriers in electrokinetic capillary electrophoresis for the separation of benzene derivatives, steroids, and phenols are shown. PMID- 11379953 TI - Synthesis and characterization of novel anionic siloxane polymers as pseudostationary phases for electrokinetic chromatography. AB - Four novel siloxane polymeric pseudostationary phases with three different ionic head groups have been synthesized and characterized for electrokinetic chromatography. Siloxane polymers are of interest in this application because of the wide range of chemistries that can be developed based on these backbones, including much of the chromatographic stationary phase chemistry developed in the last thirty years. All four of the siloxanes studied were synthesized by modification of a single methylhydrosiloxane polymer with highly acidic anionic functionalities. One of the siloxanes had both ionic groups and alkane chains attached to the siloxane backbone. The electrophoretic mobilities varied from being somewhat less than sodium dodecyl sulfate (SDS) to being much greater than SDS. The siloxanes substituted with ionic groups at all of the silicon sites showed significant nonequilibrium band broadening, severely limiting the efficiencies of these polymers. Substitution of 20% of the silicon sites with an alkyl group improved the efficiency of the separations and the peak symmetry. The chemical selectivities of the siloxane polymers are very different from SDS, but are similar to each other. PMID- 11379954 TI - Application of linear solvation energy relationships to polymeric pseudostationary phases in micellar electrokinetic chromatography. AB - Polymerized sodium 11-acrylamidoundecanoate (poly(Na 11-AAU)) was used as a pseudostationary phase (PSP) for micellar electrokinetic chromatography to separate uncharged compounds. The polymer PSP showed signifcantly different solute migration behaviors from conventional micelles including sodium dodecyl sulfate and poly (sodium 10-undecylenate), giving high separation efficiencies (>200000 theoretical plates/m). Linear solvation energy relationships were used to evaluate and characterize the chemical interactions that influence the retention behavior in the poly (Na 11-AAU) micellar system. It was found that the solute volume and solute hydrogen bond basicity mainly influenced the retention. The characteristic feature of the poly (Na 11-AAU) micellar system is that the micelle has a significantly higher capacity for dipole-dipole and dipole-induced dipole interactions as well as a slightly higher capacity for electron pair interactions than the aqueous phase. Due to its unique selectivity, the poly(Na 11-AAU) micellar system would become an attractive new option for selectivity optimization on methods development. PMID- 11379955 TI - Separation of neutral compounds by microemulsion electrokinetic chromatography: fundamental studies on selectivity. AB - The selectivity of microemulsion electrokinetic chromatography (MEEKC) was studied utilizing some uncharged model compounds like aromatic amides, steroids, and esters of nicotinic acid. The cosurfactant of the microemulsion was found to be the most important factor affecting the selectivity, and alteration between 6.6% of 1-propanol, 1-butanol, tetrahydrofuran, and 2-ethoxyethanol caused several substantial changes in the migration order. In addition, the nature of the surfactant was found to significantly affect the selectivity. In this case, changes in order of migration was observed by replacement of half the content of sodium dodecyl sulfate (SDS) with either sodium dioctyl sulfosuccinate (SDOSS), 3 (N,N-dimethylmyristylammonio) propanesulfonate (MAPS), polyoxyethylene sorbitan monolaurate (Tween 21), and polyoxyethylene 23 lauryl ether (Brij 35). MEEKC was also accomplished with 3.3% of the anionic surfactant sodium cholate and with the cationic surfactant N-cetyl-N,N,N-trimethylammonium bromide (CTMA). Both provided substantial differences in selectivity as compared to the SDS-based systems. With SDS as surfacant, the concentration was varied within 1.0-4.5%. Minor selectivity changes were observed as the concentration of the surfacant was reduced, but the major effect was a reduction in the total migration time. The organic solvent of the microemulsion droplets was found only to have minor impact on the selectivity. PMID- 11379956 TI - Towards a general approach for the impurity profiling of drugs by micellar electrokinetic chromatography. AB - A general micellar electrokinetic chromatographic (MEKC) strategy for the impurity profiling of drugs was developed involving a sodium dodecyl sulfate (SDS) and a cetyltrimethylammonium bromide (CTAB) MEKC system. With this combination, in principle, each sample component passes the detector in at least one of the two MEKC systems provided that separation buffers of the same pH are used in both systems. In order to select the proper MEKC systems, the electroosmotic flow (EOF) and micelle migration time (t(mc)) were determined for separation buffers of several pH values, containing various amounts of surfactant and organic modifier. The selectivity of the MEKC systems was studied using a mixture of compounds with a wide range of physico-chemical properties. The final selection of two adequate MEKC systems for this approach was based on the requirements that the t(mc) (i.e., analysis time) of both systems was below 20 min and that the t(mc)/t(eof) ratio was above 3 or 2 for the SDS and CTAB system, respectively. Furthermore, the systems should provide high efficiency, exhibit differences in selectivity and use moderate concentrations of modifier and surfactant, so that, if needed, further optimization is possible. The selected MEKC systems contained 60 mM SDS or 10 mM CTAB, respectively, in a phosphate buffer (pH 7.5) with 10% acetonitrile. Some test compounds with extreme mobilities were used to demonstrate the suitability of the MEKC approach to detect each component of a sample. The potential of the proposed MEKC combination for impurity profiling was demonstrated by the analysis of fluvoxamine with several impurities at the 0.1% level. PMID- 11379957 TI - Determination of carbamazepine and carbamazepine-10,11-epoxide in human serum and plasma by micellar electrokinetic capillary chromatography in the absence of electroosmosis. AB - Therapeutic drug monitoring of carbamazepine (CBZ), a widely used antiepileptic drug, is required for optimization of pharmacotherapy with this drug and for assessment of the patient's compliance to therapy. The suitability of employing micellar electrokinetic capillary chromatography (MEKC) in the absence of electroosmosis for the determination of CBZ and its main metabolite carbamazepine 10,11-epoxide (CBZE) in extracts of human serum and plasma is reported. Using micelles formed by dodecyl sulfate, analyses performed in untreated fused-silica capillaries at acidic pH and in commercially available coated capillaries under application of reversed polarity are compared. Uncoated and polyvinyl alcohol coated capillaries proved to be unsuitable for this purpose, whereas capillaries coated with linear polyacrylamide and N-acryloylaminoethoxyethanol and operated at pH 7.6 are shown to provide high-quality and reliable data on a short time scale. Assay performance is discussed via statistical analysis of the data produced from a set of quality control sera that contain up to 14 different drugs and via analysis of patient samples. Intraday and interday imprecision data for concentrations between 4.0 and 84 microM are demonstrated to be < 10%. Run times are shown to be < 50% compared to those observed in conventional MEKC at alkaline pH (i.e., in the presence of electroosmosis). PMID- 11379958 TI - Analysis of bacitracin by micellar electrokinetic capillary chromatography with mixed micelle in acidic solution. AB - A method used for quantitative analysis of bacitracin with micellar electrokinetic capillary chromatography (MEKC) is described. As capillary zone electrophoresis gave poor separation selectivity, MEKC was preferable. It was found that a zwitterionic surfactant, 3-(N,N-dimethylhexadecylammonium) propanesulfonate (PAPS) gave the best selectivity among the several surfactants studied. As the analytes tend to adsorb onto the capillary wall due to their positive charge, an acidic solution composed of Tris-phosphate buffer at pH 2.5 was necessary to diminish such adsorption. The peak tailing caused by relatively strong ion pair interaction between the analyte and PAPS micelle could be reduced by adding nonionic surfactant Brij 35 to the PAPS solution. This phenomenon is possibly explained by a mixed micelle mechanism. In order to obtain the optimal conditions and to test the method robustness, a central composite experimental design was performed. The optimal conditions are as follows: 44 cm length of fused-silica capillary with 50 microm inner diameter, 90 mM Tris-phosphate buffer (pH 2.5) containing 17 mM PAPS and 0.3% w/v-Brij 35, 18 kV applied voltage, UV detection at 192 nm and 25 degrees C column temperature. Under the optimal conditions, more than 50 peaks could be obtained in 30 min. The method had a linearity range from 1 to 0.05 mg/mL (concentration of bacitracin A). The limit of quantitation (LOQ) and limit of detection (LOD) were 0.005 and 0.0012 mg/mL, respectively. PMID- 11379959 TI - On-line coupling of partial filling-capillary zone electrophoresis with mass spectrometry for the separation of clenbuterol enantiomers. AB - The on-line coupling of capillary zone electrophoresis with mass spectrometry (CZE-MS) for the separation of enantiomers is hampered by the presence of nonvolatile chiral selectors such as cyclodextrins in the separation buffer. This problem can be overcome by use of the partial filling technique where only a part of the capillary is filled with the separation buffer containing chiral selectors. Since the electroosmotic flow is almost completely suppressed at acidic pH, that dimethyl-beta-cyclodextrin is neutral, no free cyclodextrin would reach the MS detector when using a partially filled capillary. By this method, clenbuterol enantiomers were successfully resolved and separated from salbutamol (internal standard) in aqueous solution and in plasma samples. A solid-phase extraction (SPE) was used for the preparation of plasma samples before analysis. PMID- 11379960 TI - Chiral capillary electrophoresis and nuclear magnetic resonance investigation on the structure-enantioselectivity relationship in synthetic cyclopeptides as chiral selectors. AB - In the present work, synthetic cyclohexa- and cycloheptapeptides previously singled out by a combinatorial chemistry approach have been evaluated as chiral selectors in capillary electrophoresis. By applying the countercurrent migration technique and employing a new adsorbed coating, a series of dinitrophenyl amino acids as well as some chiral compounds of pharmaceutical interest have been evaluated for enantiorecognition. The results thus obtained led to a deeper investigation of the chiral discrimination process, by carrying out nuclear magnetic resonance (NMR) studies on selected cyclopeptide-analyte complexes. These studies shed light on the chemical groups involved in the analyte-selector interaction and provided useful information for a wider application of these cyclopeptides in the separation of other drug enantiomers. PMID- 11379961 TI - Separation of dipeptide and tripeptide enantiomers in capillary electrophoresis using carboxymethyl-beta-cyclodextrin and succinyl-beta-cyclodextrin: influence of the amino acid sequence, nature of the cyclodextrin and pH. AB - The separation of the LL and DD enantiomers of dipeptides and tripeptides using cyclodextrins (CDs) containing carboxyl groups was investigated with respect to the amino acid sequence of the peptides, the nature of the cyclodextrin and the buffer pH. Compared to succinyl-beta-cyclodextrin, carboxymethyl-beta cyclodextrin was the more universal CD for enantioseparations. Reversal of the enantiomer migration order upon increasing the buffer pH from 2.5 to 3.5 was observed in some cases. As shown for Phe-Phe reversal of the migration order also occurred between pH 3.5 and 5.3. Complexation constants and complex mobilities change with pH as both, the charge of the peptide and the charge of the CD vary depending on the pH. The complexation constants and complex mobilities of the dipeptides Ala-Phe and Phe-Phe were determined in order to explain the enantiomer migration behavior in the pH range 2.5-5.3. While the complexation constants determined the migration order at pH 2.5 and 5.3, complex mobility had a strong influence around pH 3.5-3.8. PMID- 11379962 TI - Separation selectivity patterns in the capillary electrophoretic separation of anionic enantiomers by octakis-6-sulfato-gamma-cyclodextrin. AB - The capillary electrophoretic separation of anionic enantiomers with multiply charged, single-isomer, anionic resolving agents was reexamined with the help of the charged resolving agent migration model. Three general model parameters were identified that influence the shape of the separation selectivity and enantiomer mobility difference curves: parameter b, the binding selectivity (K(RCD)/K(SCD)), parameter s, the size selectivity (micro0RCD/micro0SCD), and parameter a, the complexation-induced alteration of the analyte's mobility (micro0SCD/micro0). Function analysis of the model indicates that there are six unique separation selectivity vs. resolving agent concentration patterns: in two of the patterns, separation selectivity asymptotically increases to the limiting value set by parameter b; in two other patterns, separation selectivity passes a local maximum and asymptotically decreases to the limiting value set by parameter b; and in the last two patterns, separation selectivity passes a local maximum, decreases to unity, then, after reversal of the intrinsic migration order, asymptotically increases to the limiting value set by parameter b. Though the patterns with asymptotically increasing selectivities were observed in earlier work, this paper reports the first experimental verification of the existence of the local selectivity maximum during the capillary electrophoretic separation of the enantiomers of several weak acids in high pH background electrolytes with octakis 6-sulfato-gamma-cyclodextrin as the resolving agent. PMID- 11379963 TI - Enantiomeric determination of praziquantel and its main metabolite trans-4 hydroxypraziquantel in human plasma by cyclodextrin-modified micellar electrokinetic chromatography. AB - A capillary electrophoresis method for the simultaneous quantitation of praziquantel and its main metabolite trans-4-hydroxypraziquantel enantiomers in human plasma was developed and validated using cyclodextrin-modified micellar electrokinetic chromatography. Sample clean-up involved a single-step liquid liquid extraction of plasma with toluene after the addition of NaCl. The complete enantioselective analysis was obtained in less than 7 min using 2% w/v sulfated beta-cyclodextrin as chiral selector and 20 mmol/L sodium deoxycholate as surfactant, in 20 mmol/L sodium borate buffer, pH 10. A 50 microm x 42 cm uncoated fused-silica capillary was used for the analysis, performed at a voltage of 18 kV and at 20 degrees C. The calibration curves were linear over the 125-625 ng/mL concentration range. The mean recoveries for praziquantel and trans-4 hydroxypraziquantel were up to 96 and 71%, respectively, with good precision. All four enantiomers were quantified at two concentration levels (200 and 600 ng/mL) with precision and accuracy below 15%. The quantitation limit was 50 ng/mL for ( )-(R)- and (+)-(S)-praziquantel and 62.5 ng/mL for (-)-(R)- and (+)-(S)-trans-4 hydroxypraziquantel, using 1 mL of human plasma. PMID- 11379964 TI - Simultaneous determination of disopyramide and mono-N-dealkyldisopyramide enantiomers in human plasma by capillary electrophoresis. AB - In this paper, a rapid method for the enantioselective analysis of the antiarrhythmic drug disopyramide and its main metabolite mono-N dealkyldisopyramide in human plasma by capillary electrophoresis employing the cyclodextrin-modified electrokinetic chromatography mode is described. Sample clean-up was carried out by alkalinization with sodium hydroxide followed by liquid-liquid extraction with toluene. The complete enantioselective analysis was performed within less than 5 min using 20 mmol/L sodium acetate buffer, pH 5.0, containing 0.2% w/v sulfated beta-cyclodextrin as chiral selector. A 40 cm uncoated fused-silica capillary was used for the analysis, performed at a voltage of 15 kV and at 20 degrees C. The calibration curves were linear over the concentration range of 62.5-1850 ng/mL and 125-1850 ng/mL for each enantiomer of disopyramide and mono-N-dealkyldisopyramide. The mean recoveries for disopyramide and mono-N-dealkyldisopyramide enantiomers were up to 87 and 69%, respectively. All four enantiomers studied could be quantified at three different concentrations (200, 400 and 600 ng/mL) with coefficient of variation and % relative error not higher than 15%. The quantitation limit was 62.5 ng/mL for (+) (S)-and (-)-(R)-disopyramide and (-)-(R)-mono-N-dealkyldisopyramide and 125 ng/mL for (+)-(S)-mono-N-dealkyldisopyramide, using 1 mL of human plasma. PMID- 11379965 TI - Enantioseparation of ofloxacin in urine by capillary electrokinetic chromatography using charged cyclodextrins as chiral selectors and assessment of enantioconversion. AB - A method was developed for the enantioseparation of ofloxacin, a member of the fluoroquinolones, using an anionic cyclodextrin-derivative with or without combination with a neutral cyclodextrin-derivative, as the chiral selector (s) in an electrokinetic chromatography system. The best results were obtained with 0.35 mM sulfated beta-cyclodextrin dissolved in a 50 mM phosphate buffer, pH 2.5, and at 15 degrees C. Under these conditions, a resolution of 2 was readily achieved. Furthermore, under adequate separation conditions, studies were performed in order to assess possible in vitro and in vivo enantioconversion of levofloxacin. The current method allows detection of 2 microg R-(+)-ofloxacine/mL diluted urine without the necessity of sample cleanup. PMID- 11379966 TI - Methods for the determination of binding constants by capillary electrophoresis. AB - Apparent equilibrium constants for molecular association (e.g., association constants, binding constants, dissociation constants, partition coefficients) can be determined with a variety of different capillary electrophoresis (CE) approaches. In many cases, the investigated association behavior is between a smaller molecule or ion (i.e., the solute, drug, or analyte of interest) and a larger entity (e.g., proteins, micelles, polymers, chiral selectors such as cyclodextrins, etc.). Each experimental approach has advantages and disadvantages. Frequently, it is the nature of the system being evaluated that determines the optimal experimental approach. Six different CE-based techniques for evaluating binding constants are reviewed. Examples of each method, and recent references on its use are given. PMID- 11379967 TI - Application of a green fluorescent fusion protein to study protein-protein interactions by electrophoretic methods. AB - A screening procedure for protein-protein interactions in cellular extracts using a green fluorescent protein (GFP) and affinity capillary electrophoresis (ACE) was established. GFP was fused as a fluorescent indicator to the C-terminus of a cyclophilin (rDmCyp20) from Drosophila melanogaster. Cyclophilins (Cyps) belong to the ubiquitously distributed enzyme family of peptidyl-prolyl cis/trans isomerases (PPlases) and are well known as cellular targets of the immunosuppressive drug cyclosporin A (CsA). The PPlase activity of the GFP fused rDmCyp20 as well as the high affinity to CsA remain intact. Using native gel electrophoresis and ACE mobility-shift assays, it was demonstrated that the known moderate affinity of Cyp20 to the capsid protein p24 of HIV-1 was detectable in the case of rDmCyp20 fused to the fluorescent tag. For the p24 / rDmCyp20-GFP binding an ACE method was established which allowed to determine a dissociation constant of Kd = 20+/-1.5 x 10(-6) M. This result was verified by size-exclusion chromatography and is in good agreement with published data for the nonfused protein. Moreover the fusion protein was utilized to screen rDmCyp20-protein interactions by capillary electrophoresis in biological matrices. A putative ligand of rDmCyp20 in crude extracts of embryonic D. melanogaster was discovered by mobility-shift assays using native gel electrophoresis with fluorescence imaging and ACE with laser-induced fluorescence detection. The approach seems applicable to a wide range of proteins and offers new opportunities to screen for moderate protein-protein interactions in biological samples. PMID- 11379969 TI - USDA amends regulations governing care of marine mammals and placement of confiscated animals. PMID- 11379968 TI - Michaelis-Menten analysis of bovine plasma amine oxidase by capillary electrophoresis using electrophoretically mediated microanalysis in a partially filled capillary. AB - A method for determining bovine plasma amine oxidase (PAO; EC 1.4.3.6) activity with benzylamine (Bz) as substrate is described. Electrophoretically mediated microanalysis (EMMA) combined with micellar electrokinetic capillary chromatography (MEKC) was used to perform an on-capillary enzymatic reaction and to separate the generated benzaldehyde from the other reaction products. The capillary was only partially filled with the separation solution, since the enzyme was unstable in the presence of the applied surfactant. The initial reaction velocity of the enzyme-catalyzed reaction was estimated from the peak area of the enzyme product, benzaldehyde. An amplification step was introduced by means of an on-capillary incubation of 15 min, in order to accumulate enough reaction product to detect spectrophotometrically at 254 nm. This set-up resulted in a fully automated assay, which can be carried out in less then 35 min. Using the Lineweaver-Burk equation, an average Michaelis constant (K(M)) for PAO was calculated to be 0.74 mM +/- 0.05 mM, which is consistent with previously reported values. PMID- 11379970 TI - Cognitive behavioral therapy for schizophrenia: an empirical review. AB - Early case studies and noncontrolled trial studies focusing on the treatment of delusions and hallucinations have laid the foundation for more recent developments in comprehensive cognitive behavioral therapy (CBT) interventions for schizophrenia. Seven randomized, controlled trial studies testing the efficacy of CBT for schizophrenia were identified by electronic search (MEDLINE and PsychInfo) and by personal correspondence. After a review of these studies, effect size (ES) estimates were computed to determine the statistical magnitude of clinical change in CBT and control treatment conditions. CBT has been shown to produce large clinical effects on measures of positive and negative symptoms of schizophrenia. Patients receiving routine care and adjunctive CBT have experienced additional benefits above and beyond the gains achieved with routine care and adjunctive supportive therapy. These results reveal promise for the role of CBT in the treatment of schizophrenia although additional research is required to test its efficacy, long-term durability, and impact on relapse rates and quality of life. Clinical refinements are needed also to help those who show only minimal benefit with the intervention. PMID- 11379971 TI - Are antidepressants overrated? A review of methodological problems in antidepressant trials. AB - There are no signs that the rapidly escalating use of antidepressants is reducing the burden of depressive disorders. This may be due to the fact that the evidence base for antidepressants is weaker than is commonly assumed. There are a number of methodological problems that may bias the results of clinical trials. Unblinding may inflate the response of people taking an active drug when compared with those taking an inert placebo. Modern measurement techniques may exaggerate the benefit of drug treatment. Excluding some randomized subjects from analysis may inflate the apparent effect of antidepressant drugs and publication bias means that published studies may not represent an accurate picture of the effects of treatment. In trials of long-term treatment discontinuation-related effects may masquerade as clinical efficacy. A brief survey of evidence from controlled trials does not present a consistently positive picture. Two of the largest and most reputable trials found only negligible differences between tricyclic antidepressants and placebo. The evidence on whether antidepressants are specific treatments is also inconclusive. Many other drugs not classed as antidepressants have shown positive effects in depression in controlled clinical trials. It is suggested that the interests of the pharmaceutical industry and the psychiatric profession have helped to establish the notion of the efficacy and specificity of antidepressant drugs. PMID- 11379972 TI - Qualms about balms: perspectives on antidepressants. PMID- 11379974 TI - Comparing patients' and staff members' attitudes: does patients' competence to disagree mean they are not competent? AB - The ability to process and compare the benefits and risks of a proposed treatment is considered an important component of being competent to make treatment decisions. Whether psychiatric patients' expressed treatment choice reflects their personal preferences or a deficit in their decision-making process is unclear. The aim of the study was to assess the extent to which patients and staff agree or not on various treatment-related issues. A literature search was conducted to identify published articles comparing the perceptions and attitudes of staff and patients toward various treatment issues. Twenty-eight published articles over the last 40 years were located and their main findings summarized. Analysis of the findings revealed disagreement between patients and staff in 26 of the 28 articles. The consistency of the disagreement over time and across studies suggests that the disagreement might have more to do with the fundamental difference between being a patient and a staff member rather than a patient's cognitive deficits or psychopathology. It is crucial that both patients and staff work toward building bridges when discord appears consistent and pervasive. PMID- 11379973 TI - Psychiatric outpatients report their experiences as participants in a randomized clinical trial. AB - We conducted exit interviews with 45 outpatients with severe and persistent mental illness (SPMI) who had participated in a randomized clinical trial. The interviews followed a semistructured format and were audiotaped for later transcription and rating by two independent raters. Content analyses of the interviews revealed that most participants evaluated their experiences quite favorably. For example, most noted that the assessment process was thought provoking and motivational and that the intervention groups led to increased self confidence and new friendships. Although a few participants noted that the assessment contained sensitive material, all appreciated the frequent reminders that information disclosed was strictly confidential. These results indicate that persons living with a SPMI often enjoy participating in behavioral research, which can yield immediate benefits to patient-participants. Exit interview research such as this can help investigators to understand reasons for consent and participation, to identify needs for protocol modifications, and to facilitate the integration of evidence-based interventions into the mental health care systems. PMID- 11379975 TI - Insecure attachment and alexithymia in young men with mood symptoms. AB - According to attachment theorists, affect regulation and quality of attachment are closely linked. As a personality trait associated with deficits in the cognitive processing and regulation of affects, alexithymia has been hypothesized to correlate with insecure attachment. To test this hypothesis, we studied the relationships between alexithymia, adult attachment style, and retrospective memories of separation anxiety symptoms during childhood in 100 young men with clinically significant mood symptoms. The most common DSM-IV diagnosis (N = 72) was adjustment disorder with depressed mood, with anxiety, or with mixed anxiety and depressed mood. Each participant completed the Twenty-Item Toronto Alexithymia Scale (TAS-20), the Beck Depression Inventory (BDI), the state form of the State-Trait Anxiety Index (STAI), the Attachment Style Questionnaire (ASQ), the Relationship Questionnaire (RQ), and the Separation Anxiety Symptom Inventory (SASI). Alexithymic traits were more pronounced in those participants who had patterns of insecure attachment and who reported more severe symptoms of separation anxiety during childhood, independently of the severity of their current anxiety and depressive symptoms. Among the subgroup of participants with insecure attachment styles, those with preoccupied or fearful patterns had a higher prevalence of alexithymia (65% and 73%, respectively) than those with a dismissing pattern (36%). These data suggest a role for early developmental factors in the etiology of alexithymia PMID- 11379976 TI - Gun use, attitudes toward violence, and aggression among combat veterans with chronic posttraumatic stress disorder. AB - Vietnam veterans with chronic combat-related posttraumatic stress disorder (PTSD) have been frequently reported to exhibit high levels of aggression and violent behavior. In this study, gun collection and use habits, attitudes toward violence, and self-reported levels of aggression were compared between veterans with chronic PTSD and non-PTSD veterans with equivalent histories of alcohol and substance abuse. PTSD patients reported different attitudes toward violent crime, higher levels of self-reported aggression, and a significantly higher incidence of potentially dangerous firearm-related behaviors than comparison subjects. PMID- 11379977 TI - Panic disorder with agoraphobia associated with dizziness: characteristic symptoms and psychosocial sequelae. AB - The aim of this study was to assess the prevalence of symptoms of panic disorder in a representative community sample of people with dizziness and to compare the profile of those whose panic was consistently linked to attacks of dizziness with those in whom dizziness was just one of many, variable somatic symptoms of panic. Validated questionnaires assessing physical and psychological symptoms, occupational disability, and handicap were administered to 128 people reporting dizziness in an epidemiological survey. Nearly two thirds of the sample reported having panic attacks, and one in four met key criteria for panic disorder. People whose panic symptoms were consistently associated with dizziness reported higher rates of vertigo than those with panic unrelated to dizziness, and higher rates of fainting, agoraphobic behavior, and occupational disability than either comparison group. Explanation of perceptual-motor triggers for disorientation may increase the predictability of attacks, thus reducing vulnerability to dizziness provoked panic. PMID- 11379978 TI - Consumers who call the Anxiety Disorders Association of America: characteristics and satisfaction. PMID- 11379979 TI - Features of out-of-body experiences in relation to perceived closeness to death. PMID- 11379980 TI - Is panic disorder with psychosensorial symptoms (depersonalization-derealization) a more severe clinical subtype? PMID- 11379981 TI - Locus of control and field dependence in prediction. PMID- 11379982 TI - Geriatric Depression Scale (GDS-15): a sensitive and convenient instrument for measuring depression in young anorexic patients. PMID- 11379983 TI - Dermatitis caused by radio-frequency electromagnetic radiation. PMID- 11379984 TI - Occupational erythema-multiforme-like dermatitis from sensitization to costus resinoid, followed by flare-up and systemic contact dermatitis from beta cyclocostunolide in a chemistry student. PMID- 11379985 TI - Microvascular endothelial dysfunction: a renewed appreciation of sepsis pathophysiology. AB - Severe sepsis, defined as sepsis associated with acute organ dysfunction, results from a generalized inflammatory and procoagulant host response to infection. Coagulopathy in severe sepsis is commonly associated with multiple organ dysfunction, and often results in death. The molecule that is central to these effects is thrombin, although it may also have anticoagulant and antithrombotic effects through the activation of Protein C and induction of prostacyclin. In recent years, it has been recognized that chemicals produced by endothelial cells play a key role in the pathogenesis of sepsis. Thrombomodulin on endothelial cells coverts Protein C to Activated Protein C, which has important antithrombotic, profibrinolytic and anti-inflammatory properties. A number of studies have shown that Protein C levels are reduced in patients with severe infection, or even in inflammatory states without infection. Because coagulopathy is associated with high mortality rates, and animal studies have indicated that therapeutic intervention may result in improved outcomes, it was rational to initiate clinical studies. PMID- 11379986 TI - The normal role of Activated Protein C in maintaining homeostasis and its relevance to critical illness. AB - Thrombin is a multifunctional protein, with procoagulant, inflammatory and anticoagulant effects. Binding of thrombin to thrombomodulin results in activation of Protein C and initiation of the Activated Protein C anticoagulant pathway, a process that is augmented by the endothelial cell Protein C receptor (EPCR). Activated Protein C has demonstrated antithrombotic, anti-inflammatory, and profibrinolytic properties. Its antithrombotic activity is particularly important in the microcirculation, and Protein C deficiency is associated with microvascular thrombosis. Activated Protein C has also been shown to modulate inflammation. When the level of thrombomodulin or Protein C is reduced in sepsis there is a vicious cycle of coagulation and inflammation, with potentially lethal consequences. In vitro studies and animal models have shown that Activated Protein C blunts the inflammatory and coagulant response to sepsis through a variety of mechanisms. PMID- 11379988 TI - Is exercise-induced myocardial injury self-abating? PMID- 11379989 TI - [The 115th Internal Medicine Seminar--the XVI. Vanysek Seminar--Lipids 2000 presented by the Czech and Slovak Internal Medicine Society, the Society of Czech Physicians in Brno and the Czech Atherosclerosis Society on the anniversary of the birth of Dr. Rudolf Vanysek, founder of the Brno School of Internal Medicine. 10 March 2000, Brno]. PMID- 11379990 TI - How health care works in the United States. PMID- 11379991 TI - Radiologic case study. Rectus femoris tendinitis (hydroxyapatite crystal deposition). PMID- 11379992 TI - Extra-articular wide tumor resection and limb reconstruction in malignant bone tumors at the proximal femur. PMID- 11379993 TI - Effect of insertion torque on bone screw pullout strength. AB - The effect of insertion torque on the holding strength of 4.5-mm ASIF/AO cortical bone screws was studied in vitro. Screw holding strength was determined using an Instron materials testing machine (Bristol, United Kingdom) on 55 lamb femora and 30 human tibiocortical bone sections. Holding strength was defined as tensile stress at pullout with rapid loading to construct failure. Different insertion torques were tested, normalizing to the thickness of cortical bone specimen engaged. These represented low, intermediate, high, and thread-damaging insertion torque. All screws inserted with thread-damaging torque and single cortex engaging screws inserted to high torque tightening moments showed diminished holding strength. This loss of strength amounted to 40%-50% less than screws inserted with less torque. PMID- 11379994 TI - Sacroiliac joint dysfunction: a long-term follow-up study. AB - Sixty-nine patients with sacroiliac joint dysfunction were prospectively evaluated and treated with a structured physical therapy program. Follow-up clinical outcome was obtained from a patient questionnaire administered by an independent reviewer a minimum of 2 years after treatment. Average patient age was 40 years, and 80% were women. Ninety-five percent rated their result as good or excellent, while 5% believed their outcome was fair or poor. A structured physical therapy program can produce good long-term results in most patients; however, 5% continue to be symptomatic. This small subset may be candidates for more invasive evaluation. PMID- 11379995 TI - Treatment of dysplastic and congenitally dislocated hips with the Zweymueller total hip prosthesis. AB - Uncemented Zweymueller total hip prostheses were implanted in 35 dysplastic or dislocated hips in 33 patients. Sixteen hips were dislocated and 19 hips were dysplastic; in 12 hips, an intertrochanteric or pelvic osteotomy was performed in early childhood. In all cases, the titanium screw socket was implanted at the level of the original cotyloid cavity. Osteotomy of the greater trochanter, shortening osteotomy, or roof acetabuloplasty were not performed. In cases in which the femoral cavity was too narrow for the Zweymueller stem, an anterolateral longitudinal window-shaped osteotomy was performed. In cases of severe dysplasia, cotyloid cavity bone grafts from the resected femoral head were placed medially to reinforce the acetabular bottom. Clinical and radiographic follow-up ranged from 3-8 years. Average Harris hip score improved from 47 points preoperatively to 86.2 points postoperatively. Complications included two primary anterior dislocations, two temporary femoral nerve pareses, and two deep vein thromboses. At longest follow-up evaluation, no revision was indicated in any of the hips. Satisfactory results in this series were attributed to careful patient selection, precise preoperative radiographic planning, and an operative technique that included implantation of the socket at the primary acetabulum and achievement of primary stability using press-fit fixation. PMID- 11379996 TI - Bone density of the first sacral vertebra in relation to sacral screw placement: a computed tomography study. AB - A visual three-dimensional image of the first sacral vertebra was constructed using computer software to predict the sites of strong density for better screw purchase of upper sacrum. Forty dry sacrum specimens were scanned in the prone position. An axial section, 10 mm below the S1 end plate, was selected for determining density at the region of interest. All images were stored on an optic disc and studied using the NIH Image 1.61 program. Plot analysis assessed the bone density in different regions. Also, three-dimensional pictures of the different screw paths and the related bone density in the upper sacrum were analyzed. Bone density in the anterolateral part of S1 was 115.1 +/- 10.4 pixel. Bone density for males (-99.7 +/- 11.3) was greater than for females (-131.4 +/- 9.6). Bone density in the anterolateral alar region was -108 +/- 10.6. The bone density for males (-95.6 +/- 9.8) and females (121.4 +/- 11.7) was more than the body region. Bone density in the middle anterior cortex of the ala was 759.8 +/- 11.6. Bone density for males (878.2 +/- 10.7) was greater than for females (637.6 +/- 11.9). Using surface plot, the midanterior cortex of the ala had high cortical density compared with other areas. The midanterior cortex of the sacral ala had the highest bone density. Sacral screw purchase in the midanterior cortex provides better mechanical fixation. PMID- 11379997 TI - Steroid injection and splinting in the treatment of carpal tunnel syndrome. AB - Thirty-five hands in 30 consecutive patients with carpal tunnel syndrome confirmed by nerve conduction velocity, 2-point discrimination <6 mm, and no thenar weakness were injected with corticosteroid. Patients underwent follow-up at 3, 6, 12, and 18 months or until symptoms recurred. Symptomatic relief obtained at 3 months was 34% and only 11% at 12 and 18 months. No correlation with clinical response and age, duration of symptoms, or sex was noted. PMID- 11379998 TI - Pseudotumors presenting in nonhemophiliacs. PMID- 11379999 TI - Multifocal pseudoaneurysmal bone cysts in sickle cell disease. PMID- 11380000 TI - Melorheostosis with heterotopic ossification. PMID- 11380001 TI - Pulmonary embolism after elective glenohumeral joint debridement. PMID- 11380002 TI - Congenital pseudarthrosis of the fibula. PMID- 11380003 TI - Bone morphogenetic proteins: an overview of therapeutic applications. PMID- 11380004 TI - An extant Hess incubator on display. PMID- 11380005 TI - More on urinary tract infection guidelines. PMID- 11380006 TI - Baby think it over. PMID- 11380007 TI - Caustic-induced upper airway obstruction responsiveness to nebulized adrenaline. PMID- 11380008 TI - Sedation of children. PMID- 11380009 TI - One more thought on sudden infant death syndrome. PMID- 11380010 TI - How young is too young for amblyopia screening? PMID- 11380011 TI - Where will we be in 2051? PMID- 11380012 TI - What is your diagnosis? Mid-dorsal abdominal mass. PMID- 11380013 TI - 'Battered pets': features that raise suspicion of non-accidental injury. AB - A study of veterinarians' perceptions, and experience, of non-accidental injury (NAI) to pets was undertaken using an anonymous questionnaire distributed to a random sample of 1000 small animal practitioners in the UK. NAI was acknowledged by 91.3 per cent (95 per cent confidence interval, 88.2 to 93.9 per cent) of the 404 respondents who returned questionnaires, of whom 48.3 per cent (95 per cent confidence interval, 43.4 to 53.1 per cent) had either suspected or seen NAI. Four hundred and forty-eight cases were documented, predominantly in dogs (243) and cats (182). Factors either raising suspicion, or facilitating recognition, of NAI included: implication of a particular person, features of the history, referral agency involvement, behaviour of the owner and/or the animal, nature of the injuries, and socioeconomic class of owners. Additionally, sexual abuse and suspected cases of Munchausen syndrome by proxy were recorded. PMID- 11380014 TI - Removal of oesophageal foreign bodies in dogs: use of the fluoroscopic method and outcome. AB - The case records of dogs with a diagnosis of oesophageal foreign body were reviewed and the owners of affected dogs contacted by telephone to ascertain whether there had been any long-term adverse sequelae. Case records of admissions between August 1993 and August 1998 were used. There were 65 admissions for this problem during the period. In 61 instances, forceps manipulation orally under fluoroscopic guidance was attempted to remove the foreign body. This was successful in 51 instances. Animals were followed up for a median of 24 months (minimum of four months). Three animals died or were euthanased in hospital. Two further animals died within two weeks of discharge. In the cases for which follow up was available, 42 were reported to be normal, one animal had a recurrence, one had a voice change and two had occasional coughing. Fluoroscopic-guided forceps retrieval appears to be an effective method of treatment and long-term complications are uncommon. PMID- 11380015 TI - Incomplete ossification of the humeral condyle in two Labrador retrievers. AB - Incomplete ossification of the humeral condyle (IOHC) was identified in two Labrador retrievers using computed tomography. Both dogs were non-weightbearing on the affected forelimbs. The dogs were treated by means of a bone screw placed across the humeral condyle. IOHC was originally reported in spaniel and chondrodystrophic breeds. The pathogenesis of the condition remains unknown, but may be related to impaired antebrachial bone growth, similarly to the pathogeneses of elbow dysplasia and radius curvus. PMID- 11380016 TI - Juvenile nephropathy in a Samoyed bitch. AB - A case of juvenile nephropathy is reported in a 16-week-old Samoyed bitch. Clinical, laboratory and gross postmortem findings followed by histological analysis of kidney, liver and cerebrum and transmission electron microscopy of renal tissue are described. The histological and ultrastructural findings are similar to those found in a line of related Samoyeds in Canada, termed Samoyed hereditary glomerulopathy. The case is, however, distinct from those documented in Canada as the condition is present in a young female and the mode of inheritance elucidated in Canada is one of X-linked dominance, with the disease only developing in its juvenile form in males. PMID- 11380017 TI - Hepatic cirrhosis in a five-month-old dog. AB - The clinicopathological features of an unusual case of a five-month-old male Spanish mastiff, which was presented with clinical signs indicative of severe hepatic failure, are reported. Fluid replacement therapy, colloid plasma expanders, antibiotics and diuretic drugs were unsuccessful in improving the animal's general condition, and euthanasia was elected by the owner. On necropsy, ascites and severe cirrhosis, with extensive periacinar necrosis, was found. Cirrhosis is a rare lesion in young dogs. The most common causes are circulatory disturbances, hereditary metabolic disorders or poisoning, such as aflatoxicosis and anticonvulsant therapy. The possible involvement of canine adenovirus in this case is discussed. PMID- 11380018 TI - Chronic idiopathic intestinal pseudo-obstruction in an English bulldog. AB - A case of chronic idiopathic intestinal pseudo-obstruction in an English bulldog is described. The dog was presented with chronic weight loss and vomiting. An intestinal obstruction was suspected based on clinical and radiological findings. A diagnosis of chronic idiopathic intestinal pseudo-obstruction was made on the basis of full thickness intestinal biopsies. The dog was refractory to any antiemetic therapy. Necropsy revealed marked atrophy and fibrosis of the tunica muscularis, together with a mononuclear cell infiltrate extending from the duodenum to the colon. This case was presented with clinical findings consistent with visceral myopathy in humans--namely, atony and dilatation of the whole gut- but the histological findings resembled sclerosis limited to the gastrointestinal tract. PMID- 11380019 TI - Computed tomography for the diagnosis of a lumbosacral nerve sheath tumour and management by hemipelvectomy. AB - Computed tomography was used to define the gross extent of a lumbosacral plexus nerve sheath tumour in a 10-year-old crossbred dog. The tumour was also visualised ultrasonographically, which enabled a percutaneous biopsy to be taken. Hemipelvectomy was performed to achieve a clean margin of surgical excision. PMID- 11380020 TI - Para-aural abscessation following traumatic ear canal separation in a dog. AB - Traumatic ear canal separation is rare in animals, with only eight dogs and one cat reported with the condition in the English language literature. Para-aural abscessation occurred in six of these nine animals. Diagnosis was made on otoscopic observation of a shortened, abruptly ending external ear canal that was free from advanced disease. Radiographs in those cases which have been described showed a disruption of the normal air opacity of the affected ear canal. Drainage, by creating a separate opening for the horizontal ear canal, or total ear canal ablation and lateral bulla osteotomy (TECA/LBO), have led to resolution of the clinical signs. This report adds a further case to the literature in which TECA/LBO was employed successfully. PMID- 11380021 TI - Adverse effects in veterinary practice. PMID- 11380022 TI - Is fatal Rhodococcus equi pneumonia of foals only an infection acquired by the perinate? PMID- 11380023 TI - Application of Sartwell's model (lognormal distribution of incubation periods) to age at onset and age at death of foals with Rhodococcus equi pneumonia as evidence of perinatal infection. AB - The distributions of the incubation periods for infectious and neoplastic diseases originating from point-source exposures, and for genetic diseases, follow a lognormal distribution (Sartwell's model). Conversely, incubation periods in propagated outbreaks and diseases with strong environmental components do not follow a lognormal distribution. In this study Sartwell's model was applied to the age at onset and age at death of foals with Rhodococcus equi pneumonia. The age at onset of clinical signs and age at death were compiled for 107 foals that had been diagnosed with R. equi pneumonia at breeding farms in Argentina and Japan. For each outcome (disease and death), these data followed a lognormal distribution. A group of 115 foals with colic from the University of California were used as a comparison group. The age at onset of clinical signs for these foals did not follow a lognormal distribution. These results were consistent with the hypothesis that foals are infected with R. equi during the 1st several days of life, similar to a point-source exposure. PMID- 11380024 TI - Feline vaccine-associated sarcomas. AB - Feline vaccine-associated sarcomas have presented many challenges. Initially, the etiopathogenesis and biological behavior of these tumors had to be characterized, and strategies implemented to move tumors away from problematic sites. Next, diagnostic and treatment recommendations evolved as the biologic behavior of vaccine-associated sarcomas forced early and aggressive intervention. Current therapeutic strategies are expensive, at times debilitating, and frequently fail to effect tumor control. This review summarizes the known history, epidemiology, etiology, and clinical management of vaccine-associated sarcomas after a decade of work. The next challenges must be to find more practical and effective solutions, and to eliminate the cause of vaccine-associated sarcomas. PMID- 11380025 TI - Retrospective study of orthovoltage radiation therapy for nasal tumors in 42 dogs. AB - Megavoltage radiation therapy currently is the standard of care for dogs with nasal tumors. Some studies report that surgery and adjunctive orthovoltage radiation therapy result in longer control of these tumors than does megavoltage radiation therapy alone. This study reports less effective control of nasal tumors in dogs treated with surgery and orthovoltage radiation than previously observed, supporting the superiority of megavoltage radiation therapy for these tumors. In addition, this study suggests 2 new prognostic indicators for dogs with nasal tumors and describes toxicity associated with surgery and orthovoltage therapy. Forty-two dogs with nasal tumors were treated with surgical cytoreduction and 48 Gy orthovoltage radiation therapy administered in twelve 4 Gy fractions. Median survival was 7.4 months. One- and 2-year survival rates were 37% and 17%, respectively. Dogs with facial deformity had shorter survival than those without deformity (P = .005). Dogs with resolution of clinical signs after treatment had longer survival than those with chronic nasal signs (P = .0001). Acute radiation toxicity was moderate to severe for skin and eye and negligible for oral mucosa. Toxicity healed within 1 month after radiation therapy. Late toxicity was mild, but 70% of evaluable dogs experienced persistent ocular signs. Only 39% of dogs achieved a disease-free period. PMID- 11380026 TI - Association between canine malignant lymphoma, living in industrial areas, and use of chemicals by dog owners. AB - A case-control study was carried out to determine whether residential exposure to environmental pollutants increased risk for canine lymphoma in pet dogs. One hundred one cases with cytologically or histologically confirmed lymphoma diagnosed at a veterinary teaching hospital between the middle of 1996 and the middle of 1998 were examined. Controls were obtained by choosing twice the number of dogs without neoplastic disease, with overlapping distributions of province of residence, age, sex, and breed. Information regarding animal management, residence type, professional or hobby use of chemicals by owners, and treatment with herbicides or other pesticides in the area frequently visited by the dogs was obtained with a multiple-choice questionnaire by telephone interview. Two variables were positively and independently associated with the disease, namely residency in industrial areas (odds ratio [OR]; = 8.5; 95% confidence interval [CI], 2.3-30.9) and use of chemicals by owners, specifically paints or solvents (OR = 4.6; 95% CI, 1.7-12.6). A significantly lower value of the mean age of disease onset was found in the group of dogs at risk in comparison with the group of all other dogs (6.1 +/- 0.4 years, n = 36 versus 7.5 +/- 0.4 years, n = 65, respectively; P = .008). Variables describing animal care and pesticide use were either not associated with the disease or were uninformative. We suggest that canine lymphoma may be considered a sentinel of potentially hazardous situations for humans, because of the relatively short latency between exposure and disease onset. PMID- 11380027 TI - Phase I evaluation of CCNU (lomustine) in tumor-bearing cats. AB - 1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0 9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks. PMID- 11380028 TI - Simplified methods for estimation of 99mTc-pentetate and 131I-orthoiodohippurate plasma clearance in dogs and cats. AB - The purpose of this study was to evaluate simplified methods for estimation of Technetium Tc 99m (99mTc)-pentetate and orthoiodohippurate I 131 (131I-OIH) plasma clearance in dogs and cats with 1 and 2 blood samples. Plasma clearances were calculated after a bolus injection of 1.85-11.1 MBq of 99mTc-pentetate and 131I-OIH with a 2-compartment model based on a 12-point curve as a reference method in 21 dogs and 18 cats. Three 2-sample and 3 single-sample methods were investigated. The method yielding the smallest standard deviation of the difference between the reference method and the simplified method was selected as the optimal one. Linear regression analysis was performed between the reference method and the simplified method and coefficient of determination (R2) was calculated. For 99mTc-pentetate plasma clearance, the optimal 2-sample method was the one with a mono-compartment model with samples taken at specific times. For 131I-OIH plasma clearance, the estimation was improved slightly by raising the clearance calculated with a mono-compartment model to the power of an empirically determined parameter. The optimal single-sample method was the one with a linear quadratic regression between the volume of distribution of the tracer at a specific time and the clearance calculated with 12 samples. Two-sample methods performed significantly better than did single-sample methods. The conclusion is made that 99mTc-pentetate and 131I-OIH plasma clearances can be estimated in dogs and cats with 1 or 2 blood samples with a reasonable margin of error compared to plasma clearances calculated with a 2-compartment model and 12 blood samples. PMID- 11380029 TI - Increased platelet aggregation response in Cavalier King Charles Spaniels with mitral valve prolapse. AB - Mitral valve prolapse (MVP) is a fundamental feature of myxomatous mitral valve disease in the dog. In humans, primary MVP is associated with increased platelet reactivity. In Cavalier King Charles Spaniels (CKCS), a breed predisposed to myxomatous mitral valve disease, there is a high prevalence of hypomagnesemia and platelet anomalies, such as thrombocytopenia and macrothrombocytosis. The objective of this study was to evaluate platelet aggregation responses in CKCS and to determine the relationship between the platelet aggregation response and serum magnesium concentration, MVP, mitral regurgitation (MR), and platelet count. In 19 CKCS with MVP and 7 control dogs (not CKCS), the platelet aggregation response to 3 different agonists was compared. The CKCS with >100,000 platelets/microL (n = 10) had a significantly higher maximum aggregation response with regard to all tested agonists than the CKCS with <100,000 platelets/microL (n = 9) and control dogs (n = 7). The CKCS with <100,000 platelets/microL had a platelet aggregation response similar to the control dogs. There was no correlation between degree of MVP and platelet aggregation response. Platelet diameter increased (P = .006) and serum magnesium concentration decreased (P = .04) with lower platelet concentration. In conclusion, CKCS with MVP appeared to separate into 2 groups--1 group with <100,000 platelets/microL, normal platelet aggregation, low serum magnesium concentration, and enlarged platelets, and another group with >100,000 platelets/microL, increased platelet aggregation, and normal serum magnesium concentration and platelet size. PMID- 11380030 TI - Correlation of cardiac enlargement as assessed by vertebral heart size and echocardiographic and electrocardiographic findings in dogs with evolving cardiomegaly due to rapid ventricular pacing. AB - Vertebral heart size (VHS) has been proposed as a method for quantifying cardiomegaly in dogs. This study was designed to determine how well echocardiographic and electrocardiographic findings correlated with VHS. Dogs were rapid-paced into varying degrees of cardiomegaly and were monitored by thoracic radiography, echocardiography, and electrocardiography during development of cardiomegaly. Echocardiographic and electrocardiographic parameters were compared with VHS. VHS increased with increased duration or rate of pacing or both, and left atrium-to-aorta ratio, left ventricular end-diastolic diameter, left ventricular end-systolic diameter, P wave duration, and QRS duration correlated significantly with VHS. VHS (a score obtained from routine thoracic radiographs) seems to correlate well with both echocardiographic and electrocardiographic parameters. When 9 veterinarians, experienced in interpretation of cardiac silhouettes on radiographs, measured VHS from 1 normal dog and 1 dog with severe cardiomegaly, coefficients of variation (ratio of standard deviation to the mean) for their measurements were 2.7% and 2.8%, respectively. Thus, VHS could be established with great uniformity by experienced interpreters. PMID- 11380031 TI - Transvenous embolization of small patent ductus arteriosus with single detachable coils in dogs. AB - Transvenous embolization of small patent ductus arteriosus (PDA; < or = 4 mm) with a single detachable coil was attempted in 24 dogs (median age 5.7 months, range, 2.6-65.5 months; median body weight 5.5 kg, range, 1.5-30.0 kg). Angiographic imaging of the duct and pressure measurements were made before and after embolization. The minimal ductal diameter was 2.7 +/- 0.7 mm. In all dogs, a single coil was employed regardless of residual shunting. Ten dogs (PDA minimal diameter range, 1.5-2.2 mm) received a 5-mm coil, and 14 dogs (PDA minimal diameter range, 2.9-3.6 mm) received a 8-mm coil. After coil embolization the angiographic shunt grade decreased significantly (n = 20, P < .001). Residual shunts were assessed by angiography 15 minutes after and by Doppler echocardiography 1-3 days and 3 months after the intervention. In the dogs treated with the 5-mm coils the residual shunt rate was low (0%, 10%, and 0% for angiography and Doppler echocardiography at 1-3 days and 3 months, respectively), in contrast to the dogs treated with the 8-mm coils (91%, 79%, and 67% for angiography and Doppler echocardiography at 1-3 days and 3 months, respectively). After 3 months, no residual murmur was found in dogs treated with the 5-mm coils (0/7), in contrast to murmurs in 5 of 12 (42%) dogs treated with the 8-mm coils. Despite incomplete closure in these dogs, volume loading of the left heart decreased in all dogs. Pulmonic or aortic coil embolism did not occur. Analysis of initial results shows that single detachable coil embolization is possible in all dogs with a small PDA (< or = 4 mm), but only very small PDA (< or = 2.5) could be treated effectively, and for the moderate PDA (2.6-4.0 mm) longer coils or multiple coils may be necessary to achieve complete occlusion. PMID- 11380032 TI - Practices and outcome of artificial cardiac pacing in 154 dogs. AB - Artificial pacing (AP) is a treatment for symptomatic bradyarrhythmias unresponsive to medical therapy. This retrospective study was designed to define the practices and outcome of AP in dogs at 7 referral institutions participating in the Companion Animal Pacemaker Registry and Repository (CANPACERS). The indications, implantation techniques, complications, long-term outcome, and owner satisfaction were examined. One hundred fifty-four dogs were identified as undergoing AP from January 1, 1991, to January 1, 1996. Third-degree atrioventricular (AV) block (n = 91; 59%) and sinus node dysfunction (n = 45; 29%) were the most common indications for AP Transvenous endocardial AP systems were implanted in 136 dogs (88%), and epicardial systems were implanted in 18 (12%). Complications associated with AP were reported in 84 dogs (55%). Major complications occurred in 51 dogs (33%), including dislodgement of the pacing lead (n = 15; 10%), generator failure (n = 10; 6%), cardiac arrest during implantation (n = 9; 6%), and infection (n = 7; 5%). Minor complications occurred in 47 dogs (31%), including seroma formation (n = 18; 12%), muscle twitch (n = 17; 11%), and inconsequential arrhythmias (n = 15; 10%). Fourteen dogs (9%) experienced both major and minor complications. Survival analysis revealed 1-, 2 , and 3-year survival rates of 70, 57, and 45%, respectively. Age and presence of preexisting congestive heart failure (CHF) had a negative effect on survival (P = .001). Sixty percent of dogs with preexisting CHF died within 1 year of implantation, whereas 25% of dogs without heart failure died during the same period. Owners rated their satisfaction with the procedure as high in 80% of the dogs. PMID- 11380033 TI - Muscular dystrophy in female dogs. AB - The most common form of muscular dystrophy in dogs and humans is caused by mutations in the dystrophin gene. The dystrophin gene is located on the X chromosome, and, therefore, disease-causing mutations in dystrophin occur most often in males. Therefore, females with dystrophin deficiency or other forms of muscular dystrophy may be undiagnosed or misdiagnosed. Immunohistochemistry was used to analyze dystrophin and a number of other muscle proteins associated with muscular dystrophy in humans, including sarcoglycans and laminin alpha2, in muscle biopsy specimens from 5 female dogs with pathologic changes consistent with muscular dystrophy. The female dogs were presented with a variety of clinical signs including generalized weakness, muscle wasting, tremors, exercise intolerance, gait abnormalities, and limb deformity. Serum creatine kinase activity was variably high. One dog had no detectable dystrophin in the muscle; another was mosaic, with some fibers normal and others partly dystrophin deficient. A 3rd dog had normal dystrophin but no detectable laminin alpha2. Two dogs could not be classified. This study demonstrates the occurrence of dystrophin- and laminin alpha2-associated muscular dystrophy and the difficulty in clinical diagnosis of these disorders in female dogs. PMID- 11380034 TI - Intra-abdominal dermatophytic pseudomycetoma in a Persian cat. PMID- 11380035 TI - A canine granular lymphocyte proliferative disease without an aggressive clinical course. PMID- 11380036 TI - Malignant histiocytosis in 3 cats. PMID- 11380037 TI - Systemic phaeohyphomycosis (Cladophialophora bantiana) in a dog--clinical diagnosis with stereotactic computed tomographic-guided brain biopsy. PMID- 11380038 TI - Efficacy and safety of repeated infliximab infusions for Crohn's disease: 1-year clinical experience. AB - Data from clinical trials suggest the efficacy of the chimeric tumor necrosis factor alpha monoclonal antibody infliximab in improving clinical, endoscopic, and histologic outcomes in patients with moderately to severely active Crohn's disease (CD) and fistulizing CD. To determine whether the efficacy and safety record of infliximab reported in clinical trials would be reflected in clinical use, clinical experience with infliximab was assessed in patients with CD at the University of Chicago, Chicago, Illinois. All patients with CD at this institution receiving infliximab in the first year of its release were prospectively followed up for 1 year. Disease activity was scored at the time of the initial infusion and at 1, 3, 7, and 12 weeks after infusion. Results were analyzed separately for patients with luminal or fistulous CD. Clinical response, remission, corticosteroid tapering, and adverse event data were collected. A total of 129 patients with luminal (n = 81) or fistulous (n = 48) disease received a mean of 2.38 and 3.23 infusions of infliximab per patient, respectively. After the initial infusion course, clinical response and remission rates at 3 weeks were 65% and 31% for patients with luminal disease and 78% and 24% for patients with fistulous disease, respectively. Clinical response and remission after the first infusion occurred at a median of 8 days and 9 days, respectively. In those patients who subsequently relapsed, relapses occurred after a mean of 8.5 weeks and 12.2 weeks in patients with luminal and fistulizing disease, respectively. Corticosteroid tapering was possible in > 90% of patients (luminal disease) after the initial infusion and complete withdrawal in 54% after the second infusion, with a sustained median steroid dose of 0 mg from the 4 month time-point onward. Infusion reactions or adverse events occurred in 5-13% of patients during or immediately after the initial infusion of infliximab; most were mild and easily managed and did not increase in incidence with subsequent infusions. Clinical experience with infliximab closely mirrors the findings of controlled clinical trials. Repeated administration of infliximab was efficacious and relatively well tolerated in patients with CD and demonstrated corticosteroid sparing benefits. PMID- 11380039 TI - Conventional treatment of Crohn's disease: objectives and outcomes. AB - Despite conventional medical and/or surgical intervention, endoscopic and symptomatic relapse is common among individuals with Crohn's disease (CD). Treatment goals have therefore been refocused to include achieving control of active disease and maintaining remission with agents associated with a minimum of toxic adverse effects. Conventional treatment regimens have been used with varying success in regard to these therapeutic goals. Traditionally, aminosalicylates have been considered effective in inducing a response in some patients with mild-to-moderate CD but have demonstrated little or no long-term benefit in controlled clinical trials. Glucocorticosteroid therapy is associated with higher rates of response in patients with active CD; however, clinical benefits are frequently offset by the common occurrence of corticosteroid-related toxicity. Oral controlled-release budesonide has demonstrated comparable efficacy to prednisolone with less risk for adverse effects, although many questions remain regarding the long-term use of this agent. Response to standard immunosuppressive agents such as azathioprine and 6-mercaptopurine in patients with active disease may require 3 to 6 months from initiation of treatment. These agents are therefore considered most valuable as maintenance therapy, providing consistent long-term benefit in patients with chronic refractory or corticosteroid-dependent disease. Although the incidence of allergic adverse effects is relatively low with azathioprine/6-mercaptopurine, more serious adverse effects, including bone marrow suppression, hepatotoxicity, pancreatitis, and infectious complications, can occur. Limited success in the treatment of perianal disease has been achieved with antibiotics such as metronidazole and the immunosuppressives cyclosporine and azathioprine/6-mercaptopurine. Although broader use of immunosuppressive agents has allowed improvement in the maintenance of remission in patients with CD, long-term safety data with these agents are lacking, concerns about toxicity and the potential risk for neoplasia remain, and attenuation of response with chronic immunosuppressive use can occur. Therefore, innovative therapeutic approaches are needed to meet key treatment goals often not addressed by conventional therapies. PMID- 11380041 TI - Infliximab for patients with refractory ulcerative colitis. AB - Conventional treatment options for patients with severe corticosteroid-refractory ulcerative colitis (UC) include intravenous cyclosporine, which is frequently limited by toxicity, or colectomy. The efficacy of infliximab was investigated in the treatment of 16 patients with severely active UC refractory to conventional therapy; 7 of these patients were considered for colectomy pending medical failure. All patients received a single infusion of infliximab, 5 mg/kg; 6 of 16 patients (38%) received a second infusion approximately 5 months later. Efficacy was assessed by clinical response (defined as the lack of symptoms) as well as endoscopic and histologic outcomes. Clinical, endoscopic, and histologic improvement was observed in 14 of 16 patients (88%) after treatment with infliximab. Surgery was avoided in six of seven surgical candidates (86%). Clinical remission was maintained in 14 of 16 patients (88%) for > or = 4 months, and 4 of 16 patients (25%) for 7-10 months. Most of the treated patients were completely withdrawn from corticosteroid therapy. Treatment with infliximab induced endoscopic remission at 30 days and a significant improvement from baseline in mean histologic score (p < 0.001). In conclusion, infliximab improved clinical, endoscopic, and histologic outcomes in patients with severely active UC refractory to conventional therapy, allowing corticosteroid sparing and reducing the need for colectomy. PMID- 11380040 TI - Approach to corticosteroid-dependent and corticosteroid-refractory Crohn's disease. AB - Corticosteroids are considered a drug of choice for the treatment of patients with moderately to severely active Crohn's disease (CD), an inflammatory bowel disease characterized by chronic recurrent flares of disease activity. However, among patients receiving corticosteroid therapy for induction of remission, 20% have corticosteroid-refractory disease and 36% of those with an initial response develop corticosteroid dependency within 1 year. Chronic corticosteroid exposure in patients who are corticosteroid dependent increases the risk for serious drug related adverse effects. Withdrawal or reduction of corticosteroid therapy without exacerbation of symptoms is therefore recognized as an important goal of treatment. Therapies that have been shown to facilitate "steroid sparing' include the immunomodulators azathioprine/6-mercaptopurine and methotrexate and the antitumor necrosis factor-alpha monoclonal antibody infliximab. In corticosteroid dependent patients, budesonide may be substituted for conventional corticosteroid therapy without loss of response and with less risk for toxicity, but its long term efficacy requires further evaluation. A preliminary controlled study suggests that the investigational anti-TNF monoclonal antibody CDP-571 may also be clinically beneficial as a corticosteroid-sparing agent. This review summarizes the clinical evidence that supports consideration of these agents as alternatives in patients with CD who are dependent on, refractory to, or intolerant of conventional corticosteroid therapy. PMID- 11380042 TI - Altering the natural history of Crohn's disease? AB - With the medical advances achieved in Crohn's disease (CD) over the past several years, treatment goals have expanded to include not only improvement in clinical outcomes, but also potential alteration of underlying disease processes and modification of the clinical course. A reliable prospective predictive model for the clinical course of CD is presently lacking. However, preliminary evidence suggests that the clinical expression of CD may reflect at least in part transmural and superficial mucosal inflammatory changes. Treatments that induce healing of the intestinal mucosa and submucosa may therefore provide particular clinical benefits, including sustained response or remission. As a result, endoscopic outcomes in patients with CD are increasingly included as therapeutic efficacy end points in clinical studies. Corticosteroids have been shown to rapidly relieve symptoms in most patients but generally do not improve endoscopic lesions in parallel with clinical response and are ineffective as maintenance therapy. Open-label investigations suggest that azathioprine is associated with mucosal healing; in addition, placebo-controlled studies have demonstrated that this immunosuppressive agent can provide long-term suppression of disease activity, although initial onset of clinical action is slow. The antitumor necrosis factor-alpha monoclonal antibody infliximab provides endoscopic healing in parallel with clinical improvement and also effectively maintains remission with retreatment. As the relationship between endoscopic and clinical changes in disease activity is further explored and clarified, new treatment strategies will need to be developed to improve long-term prognosis in patients with CD. PMID- 11380044 TI - Privatized child welfare services: foster parents' perspectives. AB - This study reports on the impressions of foster parents following the privatization of family foster care services in Kansas. A randomly selected sample of foster parents from one agency was surveyed to assess their current level of satisfaction, their opinions about privatization, and their experiences with newly established community-based treatment teams. Although overall satisfaction has remained high, opinions vary about specific aspects of foster parenting under managed care. Implications for supporting and enhancing the role of foster parents are discussed. PMID- 11380043 TI - Transcending conventional therapies: the role of biologic and other novel therapies. AB - Biologic and other novel therapies targeted to specific pathogenic processes offer the potential for improved treatment outcomes in patients with Crohn's disease and alteration of the course of the disease. Therapies targeted to tumor necrosis factor alpha (TNF-alpha) include anti-TNF-alpha monoclonal antibodies (infliximab and CDP-571), TNF-binding neutralizing fusion proteins (etanercept), and TNF-alpha production inhibitors (thalidomide). In placebo-controlled trials, infliximab has rapidly induced clinical response and remission in patients with moderately to severely active Crohn's disease refractory to conventional therapy and patients with fistulizing Crohn's disease, with minimal toxicity; retreatment with infliximab in patients who experienced an initial response maintained their clinical improvement. Clinical experience suggests that infliximab may also be effective when administered as corticosteroid-sparing therapy. Infliximab is the only anti-TNF-alpha therapy currently available in clinical practice for the treatment of active Crohn's disease. Controlled trials of the investigational anti-TNF-alpha agent CDP-571 show benefit for induction of clinical improvement and steroid-sparing, but further investigation is needed. A pilot study of etanercept suggested a beneficial effect, but its efficacy was not confirmed in a controlled trial. In open-label trials, thalidomide has demonstrated efficacy in patients with refractory Crohn's disease; however, the therapeutic potential of thalidomide may be severely limited by the high incidence of drug-induced side effects. Other novel agents, including anti-alpha4 integrin antibodies, interleukin (IL)-10 and IL-11, and the immunomodulators tacrolimus and mycophenolate mofetil have been evaluated as treatment in patients with severely active or fistulizing Crohn's disease in open-label and controlled trials, with varied results reported to date. The development of these new therapies is an exciting advance that promises to improve the management of Crohn's disease and expand current knowledge of underlying pathophysiologic mechanisms. PMID- 11380045 TI - Health care policies for children in out-of-home care. AB - Placement in out-of-home care is one intervention used to protect children from abuse and neglect. While children are in such care, it is the child welfare agency's responsibility to ensure that their health needs are met. The study reported here examined health care policies and services for children in 46 state child welfare agencies. Virtually all states had some sort of written policies regarding health care for children in out-of-home care. Half, however, reported having no information management system to record health care data, and only six of the 23 had computerized systems. Most states fell short of meeting the standards set by the Child Welfare League of America for the health care of children in out-of-home care. PMID- 11380046 TI - Outcomes of family-centered residential treatment. AB - This article describes a family-centered residential treatment model and presents results from a quasiexperimental study examining its effectiveness in achieving permanency outcomes for children. Greater postdischarge stability was achieved for participants in the family-centered program than in the agency's standard residential treatment service. Implications for child welfare policy and practice are highlighted. PMID- 11380047 TI - Outreach to birthfathers of children in out-of-home care. AB - This article presents findings from a study of casework outreach to birthparents of children in out-of-home care. The study explored whether the birthfather was being ignored as a resource for discharge planning. It examined the outreach and interventions of caseworkers in three New York City out-of-home care agencies. Casework activity levels were found to be higher for birthmothers than for birthfathers, and a complex relationship among the variables of gender, outreach, and response was revealed. The nature and value of more specific outreach toward birthfathers of children in care and the risk of ignoring men in the birthfamily system are discussed. PMID- 11380048 TI - Policy- and system-level approaches to health promotion in Australia. PMID- 11380049 TI - Tobacco control advocacy in Australia: reflections on 30 years of progress. AB - Australia has one of the world's most successful records on tobacco control. The role of public health advocacy in securing public and political support for tobacco control legislation and policy and program support is widely acknowledged and enshrined in World Health Organization policy documents yet is seldom the subject of analysis in the public health policy research literature. Australian public health advocates tend to not work in settings where evaluation and systematic planning are valued. However, their day-to-day strategies reveal considerable method and grounding in framing theory. The nature of media advocacy is explored, with differences between the conceptualization of routine "programmatic" public health interventions and the modus operandi of media advocacy highlighted. Two case studies on securing smoke-free indoor air and banning all tobacco advertising are used to illustrate advocacy strategies that have been used in Australia. Finally, the argument that advocacy should emanate from communities and be driven by them is considered. PMID- 11380050 TI - Slip! Slop! Slap! and SunSmart, 1980-2000: Skin cancer control and 20 years of population-based campaigning. AB - The Anti-Cancer Council of Victoria has been running sun protection programs for more than 20 years: Slip! Slop! Slap! from 1980 to 1988 and SunSmart from 1988 to the present. The Victorian Health Promotion Foundation has provided funding for the SunSmart program for the past 13 years. These programs have played an important role in changing the whole society's approach to the sun and have resulted in marked reductions in sun exposure. This article describes the social, political, economic, and organizational context within which these programs developed. Then 10 areas are discussed that illustrate a critical aspect of the development and implementation of this successful systemwide health promotion program. These areas focus on key aspects of the context within which the program operates and on issues that derive from the experience of implementing program strategies. In summary, the success of the two programs is described as having been built on two key foundations: the vital integration of research and evaluation, on one hand, and a strong basis of consistency and continuity, on the other. PMID- 11380051 TI - Collaborative intersectoral approaches to nutrition in a community on the urban fringe. AB - A case study is presented that describes the 10-year evolution of a local intersectoral project aimed at improving components of a community's food system as an approach to improving nutrition. Aspects of innovation and good contemporary practice in collaborating for health promotion are illustrated. Key initiators of the project were a university public health department, a community health service, and a local government authority. Players brought into the process included the agricultural sector and food retailers. Several strategies have contributed to the success and institutionalization of the project. These include a specific focus on organizational development and capacity building among the key intersectoral partners and the use of formative evaluation methods to hasten the natural phases of collaborative problem solving. The project achieved many policy- and system-level changes. The impact on food consumption patterns is still to be evaluated. PMID- 11380052 TI - Start right-eat right award scheme: implementing food and nutrition policy in child care centers. AB - The Start Right-Eat Right award scheme implemented in Western Australia has been used to provide the incentive to bring about improvement in food service in line with government policy and regulations in the child care industry. Theories of organizational change were used to identify processes and strategies to support the industry in translating policy into practice. A baseline survey of food service management practices, as well as process evaluation, informed action and identified barriers. Impact evaluation demonstrated that the award scheme could bring about improvements in the quality of food service; 80% of centers made changes to their menus as a result of participating. Two years postlaunch, 40% of centers have registered in the scheme. The diffusion of innovation theory is used to explain uptake and discuss results. The success of the scheme was based on four factors: an understanding of the industry, collaboration between the child care industry and government, supporting resources, and incentives. PMID- 11380053 TI - Increasing the practice of health promotion initiatives by licensed premises. AB - Licensees of all licensed premises in the Hunter Region of New South Wales, Australia, were offered free services to encourage adoption of health promotion initiatives relating to responsible service of alcohol, environmental tobacco smoke, healthy food choices, breast and cervical cancer prevention, and the prevention of HIV/AIDS. A total of 239 premises participated in the follow-up survey. Increases in prevalence ranged between 11% and 59% for alcohol-related initiatives. The prevalence of smoke-free areas and healthy food choices increased from 32% to 65% and 42% to 96%, respectively, and the provision of cancer prevention information increased from 3% to 59%. Licensed premises represent a particularly challenging sector for health promotion practitioners to work in. The results of this study suggest that the adoption of health promotion initiatives by licensed premises can be increased. A considerable opportunity therefore exists for health promotion practitioners to become more actively involved in facilitating the adoption of such initiatives in this setting. PMID- 11380054 TI - Creating smoke-free environments in recreational settings. AB - To facilitate the banning of tobacco industry sponsorship, Australian health promotion foundations were established to provide health sponsorship to sport, arts, and racing organizations. Health sponsorship dollars procure health sponsorship benefits such as naming rights, signage, personal endorsement of a (health) product by a performer or player, and structural controls such as smoke free policies. Data are presented from surveys and observations of spectators attending events sponsored by the West Australian Health Promotion Foundation (Healthway) and surveys of Healthway-sponsored organizations and the community. The results demonstrate that by using health sponsorship, Healthway increased the prevalence of smoke-free policies in recreational settings, and there was growing support for these policies. There was evidence of good compliance with smoke-free policies, thus reducing exposure to environmental tobacco smoke. The introduction of smoke-free policies in recreational settings has involved working collaboratively with sectors outside of health, taking an incremental approach to change, and gaining the support of stakeholders by communicating evaluation results. PMID- 11380055 TI - Advancing health promotion in Australian general practice. AB - Health promotion activities, while having the potential to prevent disease and decrease the burden of ill health, often play a minor role in the health care offered by general practitioners. There are several identified barriers to the involvement of Australian general practitioners in health promotion. These include structural barriers and barriers within the practice setting, individual practitioner and patient factors, and difficulties in evaluating the outcomes of health promotion activities. This article explores the barriers to the engagement of Australian general practice with health promotion and reviews several recent initiatives that have the potential to increase the health promotion activities of general practitioners. These initiatives act at the level of the individual practitioner, the practice, and in the community. Despite the lack of a coordinated national approach, these strategies form an important development in general practice. PMID- 11380056 TI - Building capacity for system-level change in schools: lessons from the Gatehouse Project. AB - The Gatehouse Project is an innovative, comprehensive approach to mental health promotion in secondary schools. It sets out to promote student engagement and school connectedness as the way to improve emotional well-being and learning outcomes. The key elements of the whole-school intervention are the establishment and support of a school-based adolescent health team; the identification of risk and protective factors in each school's social and leaning environment from student surveys; and, through the use of these data, the identification and implementation of effective strategies to address these issues. The project evaluation used a cluster-randomized controlled trial design involving 26 schools with initial results demonstrating considerable success in reducing smoking rates among Year 8 children. This article describes and accounts for how system-level changes have been made in schools through a process of capacity building. This encourages teachers, parents, and students to view the core business of education differently. PMID- 11380057 TI - Parent-managed behavioral treatment for preschool children with autism: some characteristics of UK programs. AB - Early intensive behavioral intervention for autism has attracted controversy since Lovaas (1987) reported that 47% of his experimental group attained normal functioning. We summarize child and program data from 75 children receiving EIBI in the UK. The majority of children (57%) started treatment later than in Lovaas (1987), and 16% did not exceed his minimum IQ criterion. Children experienced fewer hours of treatment (mean of 32 hours vs. 40 hours per week), and their programs received relatively infrequent supervision. 21% of programs received supervision from individuals currently accredited as competent to provide Lovaas's treatment. No child started early enough, and received 40 hours per week, and had accredited supervision. Due to these variations from his model, Lovaas (1987) findings are unlikely to be replicated for this sample of children. PMID- 11380058 TI - Effects of living room, Snoezelen room, and outdoor activities on stereotypic behavior and engagement by adults with profound mental retardation. AB - Two experiments were conducted to test the effect of a room with sensory equipment, or Snoezelen room, on the stereotypic behavior and engagement of adults with profound mental retardation. In Experiment 1, participants were observed in their living room before and after attending the Snoezelen room. Results showed that there tended to be a reduction in stereotypy and increase in engagement when participants went from their living room to the Snoezelen room, and a return of these behaviors to pre-Snoezelen levels in the living room. Positive effects in the Snoezelen room did not carryover to the living room. In Experiment 2, the living and Snoezelen rooms were compared to an outdoor activity condition with the same participants and target behaviors. Results showed that the outdoor condition was superior, the Snoezelen condition intermediate, and the living room least effective in their impact on stereotypic behavior and engagement. Conceptualizations regarding factors that maintain stereotypic behavior and engagement were discussed in the context of the three experimental conditions. PMID- 11380059 TI - The effectiveness of a group discrete trial instructional approach for preschoolers with developmental disabilities. AB - Group behavioral classroom instruction for children with developmental disabilities has been shown to allow for increased efficiency, approximation to naturalistic arrangements, and enhanced opportunity for interaction, social teaching and observational learning. This study examines the effectiveness of a group instructional extension of one to one discrete trial teaching, which involves the overlapping of trials between students along with the use of sequential and choral group teaching. A multiple baseline design across tasks was employed to examine the effectiveness of the group instructional approach in promoting acquisition of educational skills among preschoolers with autism and other developmental disabilities. A time sample interval assessment of components of the group instruction was also conducted. The approach was demonstrated to consistently increase correct responding across the task areas. Results are discussed in terms of the advantages of the group instructional approach as an adjunct to one to one discrete trial instruction. PMID- 11380060 TI - Variability in staff reports of the frequency of challenging behavior. AB - Information was collected from 88 staff about their observations of the occurrence and frequency of challenging behaviors in 22 individuals with developmental disabilities with whom they worked. Staff reports suggested considerable variability in challenging behavior in the presence of different staff and, from day to day, in the presence of the same staff. Variability was greater for stereotyped than for aggressive/destructive behavior, and for more frequent behavior. Managers reported generally less challenging behavior than their staff. The validity of the findings was discussed and their implications for research and practice considered. PMID- 11380061 TI - Analysis of outcome variables of a token economy system in a state psychiatric hospital: a program evaluation. AB - This study describes the outcome of a token economy treatment applied to 2 distinct patient populations on the same unit of a state psychiatric hospital: individuals with a dual diagnosis of mental retardation and a DSM-IV Axis I diagnosis of either (a) a severe behavior disorder (BD) or (b) a serious and persistent psychiatric disorder (PD). Results showed that patients in the PD group were more likely to complete the treatment (17/20) than those in the BD group (17/31) who were more likely to be terminated from the program (14/31). Individuals who did not complete the program were distinguished early, within the first 3 weeks of treatment. These noncompleters received significantly more fines and earned significantly fewer tokens than those who completed the program. At an average of 2.7 years post-discharge, there was no difference in the proportion of PD (12/16) and BD completers (9/11) and BD noncompleters (3/7) remaining in the community. These data show that diverse populations of patients can be treated within the same token economy program, thereby improving cost effectiveness. Future research should be directed toward characterizing those patients (e.g., BD) less likely to succeed when they enter treatment, and determining if modifications in the program can improve that outcome. PMID- 11380062 TI - Occupational contact allergy to methyldibromo glutaronitrile in abrasive cleansers and work creams. PMID- 11380063 TI - Drug treatment in pregnancy. AB - When considering drug therapy in pregnancy, the risk of treatment for the embryo/fetus has to be weighed against the risk to the mother and the child of carrying out no treatment. This is of particular relevance in certain conditions like diabetes, epilepsy or AIDS, where the risk of embryopathy is increased when no treatment is carried out and the available drugs are potentially teratogenic. However, carefully selected drugs and close meshed monitoring may even decrease the risk for the child. In many instances, unintentional drug exposure occurs in the period before the pregnancy has been diagnosed. This may lead to additional diagnostic measures or even abortion of an otherwise wanted child. In both situations, planned and unintentional drug exposure during pregnancy, insufficient information is available on the clinical conditions relevant here and the specific drugs involved. Identification of potential teratogenic effects of a new drug takes place during the early development phase. However, animal models may not be representative of specifically human characteristics, e.g. deficiences in enzymes. Since drug treatment is generally best avoided during pregnancy, pharmacokinetic studies in this population are rare. However, physiological changes, known to be relevant for some drugs do occur during pregnancy. In order to improve knowledge on the pharmacokinetics of drugs in pregnancy, population pharmacokinetic approaches may represent a solution. Intensive efforts to investigate the efficacy and safety of drugs during pregnancy are necessary. Since controlled clinical trials are usually not feasible due to ethical reasons, intensified collection of case reports as well as epidemiological studies are warranted to gain sufficient information for the counselling of pregnant women. PMID- 11380064 TI - The pharmacokinetics of cerivastatin in patients on chronic hemodialysis. AB - OBJECTIVE: The single-dose and steady-state pharmacokinetics of the HMG-CoA reductase inhibitor cerivastatin and its two major metabolites, M-1 and M-23, were evaluated in patients with renal failure on chronic hemodialysis. METHODS: After having given their informed consent, 12 end-stage renal disease patients (5 female/7 male; 18 to 63 years) received a single-dose of 0.2 mg cerivastatin sodium followed by a 4-hour dialysis session for pharmacokinetic profiling. Two to four weeks later, all patients received 0.2 mg once-daily as maintenance treatment for a period of 7 days during which PK profiling was carried out on Days 1 and 7/8, both being dialysis-free days. Plasma concentrations of parent drug and active metabolites were measured by HPLC with fluorescence detection. In addition, assessment of lipid parameters, safety and tolerability, and a complete clinical chemistry program were included in the study procedures. RESULTS: Cerivastatin was well-tolerated and no serious adverse events were observed. In spite of the short treatment period, treatment responses with respect to total cholesterol, LDL cholesterol and triglycerides lowering were observed. Mean cerivastatin and metabolite concentrations and thus systemic exposure were slightly higher (up to 50%) in patients on chronic dialysis compared to previous studies carried out in healthy subjects. The unbound fraction of cerivastatin ranged from 0.6 - 1.5% in these patients (normal range: 0.5 - 0.9%). The half lives of both parent drug (approximately 3 h) and metabolites remained unaffected and, most notably, no accumulation occurred under repeated dosing. In addition, cerivastatin clearance was not increased by concurrent dialysis as would be predicted from the high plasma protein-binding (> 99%), and there were no significant differences in cerivastatin exposure between the dialysis period and the dialysis-free profile days. CONCLUSION: Cerivastatin can be safely administered in the usual dosages to patients with end-stage renal disease on chronic hemodialysis. Based on the observed moderate increase in cerivastatin mean exposure, patients should be started at the lower end of the recommended dosing range and subsequent titration should be performed with caution. PMID- 11380065 TI - Long-term administration of the HMG-CoA reductase inhibitor lovastatin in two patients with cholesteryl ester storage disease. AB - OBJECTIVE: In order to suppress de novo cholesterol and VLDL biosynthesis, a long term therapy trial with lovastatin, a competitive inhibitor of 3-hydroxy-3 methylglutaryl-coenzyme A (HMG-CoA) reductase, was initiated in two patients with cholesteryl ester storage disease (CESD), and concentrations of plasma lipids were monitored over a period of 9 years. METHODS: We studied two male patients with enzymatically confirmed CESD in whom long-term lovastatin therapy (8 and 9 years) was begun at the age of 7 and 19 years. The diagnosis of CESD was confirmed by the measurement of human lysosomal acid lipase (hLAL) activity in cultured skin fibroblasts and leukocytes. Restriction fragment length polymorphism (RFLP) analysis revealed that both subjects are homozygotes for the common CESD splice site mutation. Levels of serum lipids and lipoproteins were measured yearly. RESULTS: During the first year, total serum cholesterol decreased from 317 to 201 mg/dl in Patient A and from 228 to 120 mg/dl in Patient B, due mainly to the reduction of low-density lipoprotein (LDL) cholesterol from 262 to 151 mg/dt in Patient A and from 166 to 66 mg/dl in Patient B. Accordingly, the LDL cholesterol : high density lipoprotein (HDL) cholesterol ratio was markedly reduced in both patients after one year of therapy. The treatment was continued and, after 9 years of further medication, low total cholesterol and LDL cholesterol levels were still maintained. CONCLUSIONS: The study demonstrates that HMG-CoA reductase inhibitors are well tolerated drugs during long-term treatment of CESD patients and may help to prevent the development of premature atherosclerosis. PMID- 11380066 TI - Does intention-to-treat analysis answer all questions in long-term mortality trials? Considerations on the basis of the ANZ trial. AB - AIMS: Intention-to-treat (ITT) analyses are the gold standard in endpoint analyses of randomized clinical trials. Valuable information, however, may be lost in this approach as the actual time on trial medication is not accounted for in patients who withdraw early. Since compliance per se can be a prognostic factor and the actual treatment time is a variable likely to influence clinical outcome, this information should be added to an ITT analysis concept. Thus, the aim is to elucidate the influence of actual treatment time on the results of ITT analysis using available data from a well-conducted, large-scale clinical trial. METHODS AND RESULTS: The ANZ trial, a carvedilol study in heart failure, is characterized by a considerable number of early withdrawals in the carvedilol group. Using the ANZ trial database, the percentage of withdrawals was calculated as well as the number of days on treatment. Fourteen out of 21 deaths (67%) in the carvedilol group occurred after discontinuation of carvedilol. At the time of death, 14 out of 29 (48%) patients in the placebo group had withdrawn from treatment. Mean time of patients on medication who withdrew and died later were 301 days for placebo, but only 204 days for carvedilol. We performed a conventional ITT analysis, and an ITT-based Cox-proportional hazards model (modified ITT) in which the actual time on trial medication was entered into the model as an explanatory variable. The risk ratio in conventional ITT analysis was 0.71 (95% confidence interval (CI) 0.41 to 1.24, p = 0.228) and 0.50 (95% CI 0.28 0.90, p < 0.001 ) in modified ITT analysis. CONCLUSIONS: Incorporation of the actual treatment time substantially influences the result of this exemplary ITT analysis. The resulting risk ratio is clearly in accordance with clinical implications and in close agreement with those of other beta-blocker trials in heart failure. Entering the actual treatment time into a Cox-proportional hazards model as an explanatory variable is reasonable if not necessary from a clinical point of view and thus may strengthen the validity of ITT analysis. PMID- 11380067 TI - Gender differences in the disposition of metronidazole. AB - OBJECTIVE: Gender is usually considered to be one of the factors influencing disposition of drugs, but the evidence available is sometimes conflicting and information for a large number of frequently used drugs is lacking. An evaluation of sex differences in the disposition ofmetronidazole was carried out during a bioequivalence study. SUBJECTS AND METHODS: Twenty-four volunteers (12 males and 12 females) were included in an open, single-dose, two-sequence, crossover randomized trial with a one-week washout interval. All volunteers received in each period, a single 250 mg dose of one of the two study formulations of metronidazole. Venous blood samples were collected immediately before and at 15 time points in an 48-hour interval after drug administration; metronidazole concentrations were determined by HPLC. Non-compartmental pharmacokinetic analysis was performed and log-transformed AUC(0-infinity) and Cmax were tested for bioequivalence. Sex differences were evaluated by means of a 4-factor (sex, sequence, treatment and period) ANOVA. RESULTS: The studied formulations were found bioequivalent according to international standards: average 90% confidence interval for AUC(0-infinity) was 98 to 104 and for Cmax 93 to 115. After correction for the administered dose/kg, AUC was about 12% lower in females than in males (p = 0.0388) and, therefore, a higher calculated oral Cl/kg was found in females. Apparent distribution volume, after correction for weight, was significantly higher in males (p = 0.0019). Metronidazole half-life and MRT were shorter in females than in males (p - 0.0014 and p = 0.0002, respectively). CONCLUSIONS: Data obtained in this study suggest that metronidazole clearance in females is about 12% higher than in males although these differences are probably of no clinical relevance. PMID- 11380068 TI - Erythrodermia induced by omeprazole. AB - OBJECTIVE: To report a case of erythrodermia that appears to be related to the intake of omeprazole (OMP) for treatment of gastroesophageal reflux disease. CASE SUMMARY: We describe a case of erythrodermia associated with OMP therapy in a 58 year-old white woman with no predisposing factors. In 1995, at the Rheumatology Outpatients' Clinic, she was diagnosed as having scleroderma (CREST subgroup) and Sjogren's syndrome associated with corticoid osteoporosis and multiple crushed vertebrae, Raynaud's disease and joint pain. In March 1998, treatment with OMP 20 mg/d p.o. was started for treatment of gastroesophageal reflux disease. She came to our hospital emergency room in October 1999 because of a severe cutaneous reaction and poor general health. The dermatology service diagnosed the erythrodermia as a skin reaction to medication. We suspected that the causative agent was OMP. Examination of a skin biopsy specimen demonstrated that it was compatible with a toxicodermic reaction to medication. Administration of OMP was suspended and the skin lesions and the patient's general state of health improved. She was discharged without symptoms. CONCLUSIONS: Our case reports, and others from the literature, suggest the importance of recognizing of the possibility of cutaneous adverse drug reactions even with medications, such as OMP, which has a good safety profile. Caution is recommended with the use of OMP, especially in elderly patients, in patients with renal insufficiency or decompensating liver disease and in patients who receive drugs that affect OMP metabolism. PMID- 11380069 TI - Relative bioavailability of two isosorbide dinitrate sublingual tablet formulations administered as single doses in healthy subjects. AB - OBJECTIVE: The purpose of this study was to evaluate the relative bioavailability and bioequivalence of a test and a reference sublingual tablet each containing 5 mg of isosorbide dinitrate in healthy volunteers. METHODS: The study was conducted as an open-label, randomized, single-dose, two-period crossover design in 20 healthy volunteers with a washout period of 7 days, under fasting conditions. Plasma concentrations of the major active metabolite isosorbide 5 mononitrate were quantified, using a validated capillary gas chromatographic assay, with electron-capture detection. The pharmacokinetic parameters used to assess the bioequivalence of the two preparations were AUC(0-infinity) and AUC(0 infinity) for the extent of absorption and Cmax and tmax for the rate of absorption. RESULTS: The calculated 90% confidence intervals of the geometric mean values of the test/reference ratios were 98.2% to 103.2% (point estimate; 100.7%) for AUC(0-infinity) 96.9% to 103.8% (point estimate; 100.3%) for AUC(0 infinity), and 87.9% to 98.2% (point estimate; 92.9%) for Cmax. No statistically significant difference was found for tmax and elimination half-life (t 1/2) values. CONCLUSION: From the results of the present study, it is concluded that the test and reference isosorbide dinitrate sublingual preparations are bioequivalent in both extent and rate of absorption and it can be assumed that they are therapeutically equivalent and exchangeable in clinical practice. PMID- 11380070 TI - Restoration of self-tolerance is a feasible approach to control ongoing beta-cell specific autoreactivity: its relevance for treatment in established diabetes and islet transplantation. AB - Major progress has been made over the last 10 years in understanding the immune mechanisms underlying autoimmune Type I (insulin-dependent) diabetes mellitus. It seems that immunotherapy is the best approach for preventing and/or arresting beta cell destruction. For immunotherapeutic approaches to be clinically applicable to Type I diabetes, they cannot rely on the use of non-specific immunosuppressants. Their relative ineffectiveness over the long term and the risk of recurrence, once the drug is withdrawn, requires permanent drug treatment which augments problems of infection and drug toxicity. One solution is to selectively tackle the pathogenic immune response. Operationally, this requires establishing immunological tolerance, i.e. a state of durable antigen-specific unresponsiveness in the absence of generalised immunosuppression. This review aims to present the rationale and clinical applicability of possible immunointervention strategies in Type I diabetes aimed at restoring self tolerance to beta cells. PMID- 11380071 TI - Genetic versus environmental aetiology of the metabolic syndrome among male and female twins. AB - AIMS/HYPOTHESIS: The aetiology of the metabolic syndrome including hyperinsulinaemia, glucose intolerance, dyslipidaemia, hypertension and obesity is not known. We studied the relative impact of genetic versus environmental factors for the development of the components in the syndrome among male and female twins. METHODS: A total of 303 elderly twin pairs participated in the study. We report concordances and heritability estimates of the components by classic twin analysis to assess the proportion of variation attributed to genetic factors. RESULTS: All components correlated significantly. The concordance rates for glucose intolerance, overall obesity and low HDL-cholesterol were significantly higher among monozygotic than dizygotic twins indicating a genetic influence on the development of these phenotypes. The heritability estimates for glucose concentration, BMI and HDL-cholesterol among monozygotic twins confirmed these findings. The heritability estimates for waist-to-hip ratio, fasting insulin and triglycerides, however, were low, indicating a major environmental influence. We found a higher genetic influence on glucose intolerance and systolic blood pressure and a lower genetic influence on low HDL-cholesterol and diastolic blood pressure among male twins compared to female twins. CONCLUSION/INTERPRETATION: Based on the correlations between the components in the syndrome, we propose a core complex including hyperinsulinaemia, obesity, hypertriglyceridaemia and low HDL-cholesterol with only weak associations to glucose concentrations and blood pressure levels. The study confirms the notion of a multifactorial aetiology of the components including genetic and non-genetic factors. The differences in aetiology between male and female twins indicate an influence of sex on several of the components in the metabolic syndrome. PMID- 11380072 TI - The expression of the p85alpha subunit of phosphatidylinositol 3-kinase is induced by activation of the peroxisome proliferator-activated receptor gamma in human adipocytes. AB - AIMS/HYPOTHESIS: Thiazolidinediones are new oral antidiabetic drugs that activate the nuclear receptor PPARgamma. Our aim was to identify potential target genes of PPARgamma in the human adipocyte in order to clarify how thiazolidinediones improve insulin sensitivity. METHODS: The effect of BRL 49653 (Rosiglitazone) on the mRNA expression of insulin receptor, insulin receptor substrate-1, p85alpha, p110alpha and p110beta subunits of phosphatidylinositol 3-kinase, Glut 4 and hormone sensitive lipase was examined in isolated adipocytes. Target mRNA levels were determined by RT-competitive PCR. RESULTS: The BRL 49653 (1 micromol/l) increased the mRNA concentrations of p85alphaPI-3 K (264 +/- 46 vs 161 +/- 31 amol/microg total RNA, p = 0.003) whithout affecting the expression of the other mRNAs of interest. This effect was dose-dependent (K0.5 = 5 nmol/l) and was reproduced by a specific activator of RXR, indicating that it was probably mediated by the PPARgamma/RXR heterodimer. The BRL 49653 also increased the amount of p85alphaPI-3K protein in adipose tissue explants (71 +/- 19%). In addition, BRL 49653 produced a more than twofold increase in insulin stimulation of phosphatidylinositol 3-kinase activity and significantly enhanced the antilipolytic action of insulin. CONCLUSION/INTERPRETATION: This work demonstrates that the gene of p85alphaPI-3K is probably a target of PPARgamma and that thiazolidinediones can improve insulin action in normal human adipocytes. Although the precise mechanism of action of BRL 49653 on PI3-Kinase activity is not completely clear, these findings improve our understanding of the insulin sensitizing effects of the thiazolidinediones, possible drugs for the treatment of Type II (non-insulin-dependent) diabetes mellitus. PMID- 11380074 TI - Dual role of interferon-gamma signalling pathway in sensitivity of pancreatic beta cells to immune destruction. AB - AIMS/HYPOTHESIS: Disruption of the interferon-gamma (IFN-gamma) signalling pathway at the level of interferon regulatory factor-1 (IRF-1) protects islets against cytokine-induced nitric oxide production and cell death in vitro. The aim of this study was to investigate the effects of a global disruption of IFN-gamma signalling, or a selective disruption of IRF-1, on beta-cell sensitivity to in vivo immune destruction. METHODS: In a first set of experiments, IFN-gamma receptor knockout mice (IFN-gammaR-/-) and interferon regulatory factor-1 knockout mice (IRF-1-/-) were rendered diabetic by injections of 50 mg streptozotocin i. p. on 5 consecutive days (MLDSTZ). RESULTS: Whereas no difference in sensitivity to MLDSTZ-induced diabetes could be observed between IFN-gammaR-/- mice and their 129/Sv/Ev controls (50% vs 55%, NS), there was an increased incidence of diabetes in IRF-1-/- mice (100% vs 67% in C57B1/6 mice, p < 0.05). A similar increased sensitivity to immune destruction of IRF-1-/- islets was observed when these islets were used as allografts. Islet graft survival rate of IFN-gammaR-/- and 129/Sv/Ev islets, when transplanted in alloxan-diabetic BALB/c recipients, was comparable (12.0 +/- 1.9 days vs 12.9 +/- 2.3 days, NS). Allograft rejection, however, of IRF-1-/- islets by BALB/c recipients occurred more rapidly than following transplantation to their C57B1/6 controls (9.1 +/- 2.0 days vs 13.1 +/- 1.5 days, p < 0.003). CONCLUSIONS/INTERPRETATION: These data indicate that IFN-gamma signal transduction at the beta-cell level is not essential for immune beta-cell destruction in vivo. Moreover, disruption of the IRF-1 gene in pancreatic islets increases susceptibility to beta-cell killing, suggesting that IRF-1 might be necessary for the expression of putative beta-cell "defence and/or repair" genes. PMID- 11380073 TI - The effects of brain-derived neurotrophic factor on insulin signal transduction in the liver of diabetic mice. AB - AIM/HYPOTHESIS: We previously reported that repeated subcutaneous or intracerebroventricular injection of brain-derived neurotrophic factor (BDNF) reduces blood glucose concentrations in obese diabetic C57BL/KsJ-db/db mice. In this study, we assessed the effects of BDNF on insulin action in peripheral tissues of diabetic mice. METHODS: First, brain-derived neurotrophic factor (20 mg/kg) was subcutaneously given to male db/db mice for 14 days and then the insulin-stimulated tyrosine phosphorylation of insulin receptors and insulin stimulated phosphatidylinositol (PI) 3-kinase activity in peripheral tissues was assessed. Second, we examined the effects of a single subcutaneous or intracerebroventricular brain-derived neurotrophic factor injection on insulin responsiveness in liver and skeletal muscle of streptozotocin (STZ)-induced diabetic mice. Third, the effects of brain-derived neurothrophic factor on insulin action were also examined in cultured cells. RESULTS: Repeated injection of BDNF to db/db mice for 14 days enhanced insulin-stimulated tyrosine phosphorylation of insulin receptors in liver and insulin-stimulated PI 3-kinase activity in liver, skeletal muscle and interscapular brown adipose tissue. We then examined the rapid effect of BDNF on insulin signalling in vivo. A single subcutaneous or intracerebroventricular injection of BDNF rapidly increased insulin-stimulated tyrosine phosphorylation of insulin receptors and PI 3-kinase activity in liver of STZ-mice. No direct effect of brain-derived neurothrophic factor was observed on insulin signalling in primary cultured hepatocytes, L6 muscle cells or 3T3-L1 adipocytes. Brain-derived neurothrophic factor did not affect either glucose uptake or gluconeogenesis in these cells. CONCLUSION/INTERPRETATION: These data indicate that brain-derived neurothrophic factor rapidly enhances insulin signal transduction in liver and shows hypoglycaemic action in diabetic mice. PMID- 11380075 TI - Association between islets of Langerhans and pancreatic ductal system in adult rat. Where endocrine and exocrine meet together? AB - AIMS/HYPOTHESIS: Studies on the functional and morphological relations between exocrine and endocrine pancreas have been conducted mainly to disclose the influence of islets of Langerhans on acinar parenchyma. Less attention has been paid to the relations occurring between islets and pancreatic ducts. METHODS: A series of consecutive sections of normal adult rat pancreas were double stained with islet (hormones) and duct (cytokeratin 20) markers. Electron microscopy was conducted to investigate the ultra-structural features of duct-islet relations and anti-insulin immunogold labelling was carried out to reveal the presence of insulin in the pancreatic duct system. RESULTS: Consecutive double-stained sections demonstrated that 73.60 +/- 2.97% of the islets were attached to the ducts. For each series, 93.48 +/- 5.43 % of the islets contacting the duct tree were associated with small-sized ducts or centroacinar cells. Electron microscopy revealed that some insulin and somatostatin cells do face the duct lumen. Insulin was detected within the duct lumen and in the endosomal compartment of the duct cells. CONCLUSIONS/INTERPRETATION: The finding that most islets are connected with the duct system in the adult pancreas is discussed in terms of hormone secretion into the ducts, islet histogenesis and the relation among the three tissue components of the pancreas, the endocrine, the exocrine and the duct system. PMID- 11380076 TI - Effect of type II diabetic peripheral neuropathy on gait termination in the elderly. AB - AIMS/HYPOTHESIS: Healthy elderly people can have difficulties in precisely terminating gait due to age-related decline. Diabetes mellitus accelerates the neurodegenerative process, which results in an additional decline in motor control. This biomechanical study investigated goal-oriented gait termination in healthy elderly and elderly diabetic subjects. The trajectories of the centre of pressure and the centre of mass during the gait termination process were analysed in particular. It was hypothesised that the pathology results in an unstable gait termination, expressed in larger overshoots of the centre of pressure and the centre of mass than in healthy control subjects. METHODS: A total of 15 subjects with Type II (non-insulin-dependent) diabetes mellitus with impaired foot sensitivity due to polyneuropathy (median, 66 years) were matched according to age, gender and body mass index with 15 healthy elderly subjects (median, 67 years). The participants walked at their own pace along the walkway and stopped in front of a marked stopping line while kinetic and kinematic data were recorded. RESULTS: The diabetic subjects approached the stopping line more slowly (p = 0.002) than the healthy elderly subjects. They also exhibited a weaker maximal braking force (p = 0.011) and a prolonged relative time to develop this force (p = 0.023). Despite this slower motion, the centre of pressure overshoots were larger in the diabetic subjects than in the healthy elderly (p = 0.027). CONCLUSION/INTERPRETATION: The results show differences between healthy elderly and diabetic subjects during easy goal-oriented stopping tasks. Changes in gait termination parameters and the increased overshoots in particular document the pathology-related decline in postural stability. PMID- 11380077 TI - Endothelium-dependent and independent vasodilation studies at normoglycaemia in type I diabetes mellitus with and without microalbuminuria. AB - AIMS/HYPOTHESIS: We examined whether endothelial function is impaired in patients with Type I (insulin-dependent) diabetes mellitus under conditions of near normoglycaemia compared with age-matched healthy control subjects. Our aim was to determine whether microalbuminuria is associated with endothelial dysfunction in Type I diabetes. METHODS: Endothelial function, measured as post-ischaemic flow mediated dilatation of the brachial artery using ultrasound, was compared among 17 microalbuminuric and 17 normoalbuminuric diabetic patients, and 17 control subjects. Glyceryl trinitrate-mediated dilatation of the brachial artery was used to measure endothelium-independent function. All diabetic patients were studied at near-normoglycaemia, using insulin and 5 % dextrose infusions to maintain blood glucose between 3.5 and 8.0 mmol/l. RESULTS: Flow-mediated dilatation was significantly lower in microalbuminuric diabetic patients (3.2 +/- 0.3%) compared with normoalbuminuric diabetic patients (5.4 +/- 0.6%) and control subjects (7.9 +/- 0.6%, p < 0.001). Normoalbuminuric diabetic patients also had significantly lower flow-mediated dilatation than control subjects (p = 0.01). Glyceryl trinitrate mediated dilatation was significantly lower in the microalbuminuric patients compared with the control subjects (11.9 +/- 1.1% vs 20.0 +/- 1.2%, p = 0.001). Albumin excretion rate and glycated haemoglobin showed a significant negative independent correlation with flow-mediated dilatation (both p < 0.05). CONCLUSION/INTERPRETATION: Type I diabetic patients show endothelial dysfunction at near-normoglycaemia compared with the control subjects, and this abnormality is more marked in diabetic patients with microalbuminuria. Endothelial dysfunction in Type I diabetes is related to the albumin excretion rate and glycaemic control. The presence of endothelial dysfunction in normoalbuminuric diabetic patients suggests it could precede microalbuminuria as an early risk marker for cardiovascular disease. PMID- 11380078 TI - Haptoglobin phenotype and diabetic nephropathy. AB - AIMS/HYPOTHESIS: To determine if the haptoglobin 2 allele is associated with an increased risk for the development of diabetic nephropathy. METHODS: This study included 110 consecutive normotensive subjects with Type I (insulin-dependent) diabetes mellitus and Type II (non-insulin-dependent) diabetes mellitus seen in two outpatient clinics in Israel. Diabetes duration was greater than 10 years for Type I diabetes and more than 5 years for Type II diabetic subjects. Microalbuminuria was defined as urinary protein excretion of 30 to 300 mg/24 h, and macroalbuminuria was defined as urinary protein excretion of greater than 300 mg/24 h. Serum was taken from subjects for haptoglobin typing by gel electrophoresis. RESULTS: Of the participating subjects 54 had Type I and 56 had Type II diabetes. None (0/18) of the subjects homozygous for the haptoglobin 1 allele (1-1) showed any sign of diabetic nephropathy, as compared with 34 % (19/55) of subjects homozygous for the haptoglobin 2 allele (2-2) and 27 % (10/37) of heterozygous subjects (2-1) (p < 0.04). Of the subjects 29 showed macroalbuminuria. The risk of developing macroalbuminuria was found to be greater in subjects with two haptoglobin 2 alleles (22 %) (12/55) as compared with one haptoglobin 2 allele (8 %) (3/37) or no haptoglobin 2 alleles (0%) (0/18) (p < 0.03). CONCLUSION/INTERPRETATION: By showing a graded risk relation to the number of haptoglobin 2 alleles in Type I and Type II diabetic subjects, these studies further support our hypothesis that the haptoglobin phenotype is a major susceptibility gene for the development of diabetic nephropathy. PMID- 11380079 TI - Oxidative stress impairs insulin internalization in endothelial cells in vitro. AB - AIMS/HYPOTHESIS: Because oxidative stress has been suggested to be a significant contributing factor in the development of endothelial dysfunction and insulin resistance, we investigated whether reactive oxygen species contribute to insulin resistance by impairing insulin uptake through an effect on endothelial insulin receptor function. METHODS: Following a 2-h pro-oxidant challenge with xanthine oxidase, we examined the temporal pattern of insulin processing in the human umbilical endothelial cell line Ea.Hy926 and bovine aortic endothelial cells equilibrated with [125I]-insulin. Insulin receptor mRNA concentrations were analysed by RT-PCR and insulin receptor tyrosine phosphorylation and protein concentrations were estimated by western blotting. RESULTS: Xanthine oxidase exposure resulted in a major reduction in total insulin receptor-mediated [125I] insulin internalization over a 1-h period in both Ea.Hy926 and bovine aortic endothelial cells. After 15 min, untreated bovine aortic endothelial cells internalized fivefold more cell-bound [125I]-insulin than pro-oxidant treated cells. The [125I]-insulin disappeared from the cell surface at a similar rate in both pro-oxidant and untreated cells, with relatively more [125I]-insulin being released into the medium in pro-oxidant treated cells. Although xanthine oxidase reduced insulin receptor mRNA and protein concentrations, cell surface insulin binding capacity was not affected. Following 5 min insulin exposure, insulin receptor auto-phosphorylation was considerably reduced in cells challenged with xanthine oxidase for 2 h, which could be important for insulin receptor activation and internalization. CONCLUSION/INTERPRETATION: Oxidative stress impairs insulin endocytosis in both arterial and venous endothelial cell lines. This was not a consequence of modified insulin binding capacity but could involve insufficient insulin receptor activation. PMID- 11380080 TI - Saturated non-esterified fatty acids stimulate de novo diacylglycerol synthesis and protein kinase c activity in cultured aortic smooth muscle cells. AB - AIMS/HYPOTHESIS: Insulin resistance is linked with a cluster of multiple risk factors and excessive acceleration of atherosclerosis. The underlying mechanism is not, however, fully understood. METHODS: To determine the link between insulin resistance and altered vascular function, we focused on the effect of various non esterified fatty acids on diacylglycerol-protein kinase C pathway and mitogen activated protein kinase activity in cultured aortic smooth muscle cells. RESULTS: Incubation of the cells with saturated non-esterified fatty acids (200 micromol/l) for 24 h, such as palmitate or stearate, induced a significant increase in diacylglycerol concentrations by about fivefold or eightfold, respectively, whereas oleate induced a slight increase in diacylglycerol concentrations by 1.8-fold and arachidonate induced none. In addition, the increased diacylglycerol concentrations induced by palmitate were completely restored to control concentrations by triacsin C, acyl-CoA synthetase inhibitor. These results suggest that saturated non-esterified fatty acids may increase diacylglycerol concentrations through de novo pathway by stepwise acylation. In parallel with the increased diacylglycerol, incubation of the cells with saturated non-esterified fatty acids significantly induced the activation of protein kinase C and mitogen-activated protein kinase. The palmitate-induced increase in mitogen-activated protein kinase activity was restored to control concentrations by GF109203X (5 x 10(-7) mol/l), a specific protein kinase C inhibitor, suggesting a protein kinase C-dependent activation of mitogen activated protein kinase. CONCLUSION/INTERPRETATION: Saturated non-esterified fatty acids induced an increase in de novo diacylglycerol synthesis and subsequent activation of protein kinase C and mitogen-activated protein kinase in cultured aortic smooth muscle cells. This could contribute to the altered vascular functions in the insulin resistant state. PMID- 11380081 TI - Effects of an extracellular metal chelator on neurovascular function in diabetic rats. AB - AIMS/HYPOTHESIS: Increased oxidative stress has been causally linked to diabetic neurovascular complications, which are attenuated by antioxidants. There are several possible sources of reactive oxygen species in diabetes. Our aim was to assess the contribution of free radicals, produced by transition metal catalysed reactions, to early neuropathic changes. To this end, we examined, firstly, the effects of an extracellular high molecular weight chelator, hydroxyethyl starch deferoxamine, which is expected to be confined to vascular space, on nerve perfusion and conduction deficits in diabetic rats and, secondly, the action of a single chelator dose. METHODS: Diabetes was induced by streptozotocin. In vivo measurements comprised sciatic nerve motor conduction velocity and endoneurial perfusion, monitored by hydrogen clearance microelectrode polarography. RESULTS: We found that 8 weeks of diabetes reduced sciatic blood flow and conduction velocity by 48.3 % and 19.9% respectively. Two weeks of intravenous treatment corrected these deficits. Starch vehicle was ineffective. The time-course of action of a single hydroxyethyl starch-deferoxamine injection was examined in diabetic rats. There was a rapid increase in nerve blood flow on day 1, which remained within the non-diabetic range for 9 days before declining to the diabetic level at day 27. In contrast, conduction velocity changes were slower, reaching the non-diabetic range at day 6 and declining to the diabetic level at day 27. CONCLUSION/INTERPRETATION: Extracellular transition metal catalysed reactions play a major role in the neurovascular deficits of experimental diabetes. Given the long-lasting effect of a single treatment, extracellular metal chelator therapy could be suitable for further assessment in clinical trials. PMID- 11380082 TI - Common variants in the beta2-(Gln27Glu) and beta3-(Trp64Arg)--adrenoceptor genes are associated with elevated serum NEFA concentrations and type II diabetes. AB - AIMS/HYPOTHESIS: Higher NEFA concentrations predict Type II (non-insulin dependent) diabetes mellitus but it is not known whether higher NEFA concentrations are genetically determined or reflect coexisting obesity. To address this question we studied whether common variants in two genes encoding for key regulators of lipolysis, the beta2- and beta3- adrenoceptors (B2AR and B3AR) are associated with NEFA concentrations and Type II diabetes. METHODS: A total of 1054 Swedish subjects with varying degrees of glucose tolerance were genotyped for the Gln27Glu variant in the B2AR and for the Trp64Arg variant in the B3AR genes using PCR-RFLP. RESULTS: The B2AR Gln27 allele was more frequent in 219 Type II diabetic patients than in 237 non-diabetic subjects (59.8 % vs 52.3 %; OR = 1.72, p = 0.02) while there was no significant difference in the frequency of the B3AR Arg64 allele. Subjects homozygous for the protective alleles (Glu27 and Trp64) had, however, a lower prevalence of diabetes than subjects with other genotype combinations (OR = 0.58, p = 0.03). Among sibling pairs discordant for the B2AR Gln27Glu polymorphism, siblings with an excess of the Gln27 allele had higher fasting insulin (n = 217; p = 0.02) and NEFA concentrations (107 sex-matched pairs; p = 0.01) than siblings with an excess of the Glu27 allele. Among sibling pairs discordant for the B3AR Trp64Arg variant, siblings with the Arg64 allele had higher 2 h glucose (n = 48; p = 0.01) and NEFA concentrations (16 pairs matched for sex; p < 0.04) than siblings with the Trp64Trp64 genotype. CONCLUSIONS/INTERPRETATION: Common variants in the beta2- and beta3- adrenoceptor genes are associated with increased fasting insulin and NEFA concentrations and could increase susceptibility to Type II diabetes. PMID- 11380083 TI - Aquaporin-2 and urea transporter-A1 are up-regulated in rats with type I diabetes mellitus. AB - AIMS/HYPOTHESIS: Although the urine flow rate is considerably higher in diabetes mellitus, water reabsorption is greatly increased to concentrate an increased amount of solutes. Our study evaluated the expression of aquaporins and urea transporters, which are essential to the urinary concentration process. METHODS: Northern blot and immunoblot were used to quantify mRNA and proteins for aquaporin-2 (AQP2) as well as urea transporters UT-A1, UT-A2 and UT-B1, in subzones of the renal medulla of rats with streptozotocin-induced diabetes. RESULTS: In these rats, glycaemia, urine flow rate and water reabsorption were respectively fourfold, nine-fold and fourfold those of control rats. The AQP2 protein isoforms were significantly up-regulated in outer and inner medulla. In the base and tip of inner medulla, UT-A1 mRNA was significantly up-regulated (three- and 1.3-fold, respectively) as well as the 117 kD protein (ten- and threefold, respectively) whereas the 97 kD protein was not changed or decreased twofold, respectively. This suggests that, in diabetes, the inner medullary collecting duct is endowed with more UT-A1, especially in its initial part. In the case of mRNA and proteins of UT-A2, located in thin descending limbs in the inner stripe of outer medulla, they were respectively not changed and down regulated in diabetic rats. CONCLUSION/INTERPRETATION: This study shows that in diabetes, the increased expression of AQP2 and UT-A1 in medullary collecting duct is consistent with an improved concentrating activity. In addition, the underexpression of UT-A2 and the overexpression of UT-A1 in the initial medullary collecting duct are reminiscent of the changes seen after experimental reduction of urine concentration or low protein feeding. PMID- 11380085 TI - Tumour necrosis factor-alpha (TNF-alpha) polymorphisms-857C/A and -863C/A are associated with TNF-alpha secretion from human adipose tissue. PMID- 11380084 TI - C-peptide responses after meal challenge in mice transplanted with microencapsulated rat islets. AB - AIMS/HYPOTHESIS: This study aimed to assess a response of microencapsulated rat islets to a meal challenge after being transplanted intraperitoneally into diabetic mice. METHODS: Microencapsulated rat islets or control naked syngeneic mouse islets were transplanted intraperitoneally into mice with streptozotocin induced diabetes. Meal challenges were done 3, 6 and 9 weeks after transplantation. Glucose-induced insulin secretion from microencapsulated islets before and after transplantation was assessed in vitro. RESULTS: Within the first week, all animals transplanted with either microencapsulated rat islets or with syngeneic murine islets became normoglycaemic (< 11 mmol/l). At 4 and 6 weeks, body weight was less than normal in the non-diabetic control mice. Mice with the encapsulated rat islets had lower fasting glucose concentrations and more rapid glucose clearance after a meal challenge than the control mice. The group of mice with transplanted syngeneic islets had similar glucose profiles to control mice, except for slightly accelerated glucose clearance. The C peptide responses of mice with either microencapsulated or naked islets were clearly lower than the controls. An increase of C peptide appeared as early as 20 min in the plasma of the group with encapsulated islets, but this was considerably slower than in the other two groups. Microencapsulated rat islets retrieved 9 weeks after transplantation did not lose their ability to respond to glucose, but their output was less than half of the pretransplant control islets. CONCLUSION/INTERPRETATION: The delivery of C peptide and presumably the accompanying insulin are delayed by restrictions of the capsules and the peritoneal location. However, this delay in reaching peripheral target organs does not prevent microencapsulated grafts from efficiently clearing glucose after a meal. PMID- 11380086 TI - Mastix is another allergen causing bone-setter's herbs dermatitis. PMID- 11380087 TI - Closed mitral commissurotomy: in defense of an 'old-fashioned' procedure. PMID- 11380088 TI - Closed commissurotomy versus balloon valvuloplasty for rheumatic mitral stenosis. AB - BACKGROUND AND AIM OF THE STUDY: Closed mitral commissurotomy (CMC) and percutaneous mitral balloon valvuloplasty (PMBV) were compared by their initial results and Doppler echocardiographic data obtained at one week and one year after the procedure. METHODS: Of 580 patients with severe rheumatic mitral stenosis, 280 underwent CMC and 300 PMBV. The mean pre-procedural transmitral gradient (TMG) was 21 +/- 6 mmHg in the CMC group and 20 +/- 5 mmHg in the PMBV group (p = 0.6); the mean mitral valve area (MVA) was 1.1 +/- 0.2 cm2 in both groups. RESULTS: Mortality was 0.7% after CMC and 0.3% after PMBV; the primary success rates were 98.3% and 89% respectively (p <0.0001). Two CMC patients and three PMBV patients underwent emergency mitral valve replacement. At the first week, the mean TMG was decreased to 4 +/- 3 mmHg in the CMC group, and to 5.8 +/- 2 mmHg in the PMBV group (p <0.0001). The mean MVA was increased to 2.5 +/- 0.5 cm2 after CMC, and to 2.1 +/- 0.4 cm2 after PMBV (p <0.0001). After one year, TMG was 5.4 +/- 4 mmHg in the CMC group (p <0.0001) and 7.1 +/- 3 mmHg in the PMBV group (p <0.0001); MVA was 2.3 +/- 0.5 cm2 (p <0.0001) and 1.9 +/- 0.4 cm2 (p <0.0001), respectively. The results of CMC were significantly better (p <0.0001) with regard to TMG and MVA at these times. A significant decrease was also seen in mean left atrial diameter and pulmonary artery pressure in both groups (p <0.0001). CONCLUSION: Although satisfactory results can be achieved using either approach, CMC provides a higher primary success rate, greater MVA augmentation, and better technical control during the procedure, while reducing the cost. PMBV shortens in-hospital stay and eliminates the risk imposed by thoracotomy and anesthesia. Therefore, in our practice, when surgical intervention is contraindicated due to associated problems, PMBV may be the preferred approach, but exposure to radiation may be of concern in pregnant patients. PMID- 11380089 TI - Role of closed mitral commissurotomy in mitral stenosis with severe pulmonary hypertension. AB - BACKGROUND AND AIMS OF THE STUDY: Closed mitral commissurotomy (CMC) is a well established method for treatment of rheumatic mitral stenosis, but outcome in patients with severe pulmonary arterial hypertension (PAH) has not been clearly documented. METHODS: Between April 1996 and October 1999, among 61 patients who underwent CMC, 27 had severe PAH (systolic pressure > 100 mmHg). Of these patients, 11 were in NYHA class III, and 16 were in class IV. Preoperatively, the mean pulmonary artery (PA) pressure was 107.85 +/- 5.74 mmHg (range: 100-118 mmHg), mitral valve area (MVA) 0.704 +/- 0.106 cm2 (range: 0.5-0.91 cm2), and transmitral gradient 11.93 +/- 1.54 mmHg (range: 10-15 mmHg). The echocardiographic mitral valve score was 6.37 +/- 1.11 (range: 6-10). RESULTS: There was no operative mortality or incidence of significant (> or = 2+) post-CMC mitral regurgitation or cerebrovascular accident. The MVA increased to 2.385 +/- 0.248 cm2 (range: 1.9-2.8 cm2), the transmitral gradient fell to 2.44 +/- 0.51 mmHg (range: 2-3 mmHg), and postoperative PA systolic pressure fell to 33.33 +/- 8.20 mmHg (range: 30-60 mmHg). During a mean follow up of 26.9 months (range: 11 51 months), 23 patients were in NYHA class I and four were in class II. There were no significant differences in parameters between sexes, but mean male age was five years less than mean female age. CONCLUSION: In the subset of patients with severe PAH, surgical CMC is a safe and effective procedure that results in greater MVA and a more significant and sustained fall in PA pressure compared with reported series of percutaneous balloon mitral valvuloplasty. PMID- 11380090 TI - Early beneficial effect of preservation of papillo-annular continuity in mitral valve replacement on left ventricular function. AB - BACKGROUND AND AIM OF THE STUDY: Impairment of left ventricular (LV) function after mitral valve replacement (MVR) has been the most important factor to determine morbidity and mortality. With this in mind, LV performance in the postoperative period was assessed with and without preservation of papillo annular continuity in MVR. METHODS: Between March 1994 and August 1998, a total of 383 valve prostheses (202 MVR, 65 AVR, 58 MVR+AVR) were implanted in 325 patients, 177 of whom underwent MVR with Starr Edwards ball cage prostheses (the study group). Of these 177 patients, 105 had MVR with preservation of the posterior mitral leaflet (group I), and 72 had conventional MVR (group II). Predominant lesions were mitral stenosis in 81, mitral regurgitation in 42, and mixed mitral lesion (MS/MR) in 54. Concomitant tricuspid valve annuloplasty was performed in 13, and atrial septal defect repair in five. Sixteen patients underwent MVR for mitral restenosis. In-vivo performance of the prostheses and LV function was evaluated by M-mode and Doppler echocardiography. RESULTS: At 3-6 months clinical improvement was seen in NYHA class, with reduction in cardiothoracic ratio among patients with preserved papillo-annular continuity, irrespective of lesion type. Significant reductions (p <0.05) were seen in left atrial dimensions (54.10 +/- 8.79 preop. versus 44.64 +/- 8.54 postop.; p <0.05), left ventricular end-diastolic dimensions (LVEDD) (50.84 +/- 10.42 preop. versus 41.21 +/- 7.16 postop.; p <0.05) and end-systolic dimensions (LVESD) (34.76 +/- 7.94 preop. versus 28.81 +/- 5.79 postop.; p <0.05) in patients who had their posterior mitral leaflet preserved with significant improvement in ejection fraction (60.31 +/- 8.22 versus 64.47 +/- 7.93; p <0.05). Further analysis of data in group I patients showed significant reductions in left atrial dimensions, LVESD and peak gradient, along with improved ejection fraction compared with conventional (group II) patients. CONCLUSION: Deterioration in LV function in patients undergoing conventional MVR indicates chordal resection as a putative mechanism. This study supports the concept that maintenance of continuity between the mitral annulus and papillary muscles has a beneficial effect on postoperative LV function, and is particularly important in patients with mitral stenosis with depressed preoperative LV systolic function. PMID- 11380091 TI - Echocardiographic correlates of left ventricular outflow obstruction and systolic anterior motion following mitral valve repair. AB - BACKGROUND AND AIM OF THE STUDY: Systolic anterior motion (SAM) of the mitral valve resulting in left ventricular outflow obstruction is a well-recognized complication of repair of the degenerative myxomatous mitral valve. A precise mechanism is unknown. A current approach consists of sliding annuloplasty of the posterior leaflet. It was postulated that excess tissue of the anterior mitral leaflet (AML) was as equally (or more) important as the excess posterior mitral leaflet (PML) tissue in the development of SAM subsequent to valve repair. METHODS: Thirty-two patients without post-repair SAM (No-SAM group) were compared with eight patients with SAM (SAM group). The AML and PML heights and the mitral annulus diameter were measured by TEE using mid-esophageal four-chamber and long axis planes. RESULTS: Pre-repair TEE showed the AML height to be greater in the SAM group (p = 0.04), and that of the posterior leaflet tended to be greater (p = 0.08), whilst the annular dimensions were similar in both groups. In the post repair status, the AML height was markedly greater (p = 0.005) and the annulus markedly smaller (p = 0.001) in the SAM group. Post-repair assessment showed the relative difference between AML height and annular dimension (AML - Ann) as well as the difference between combined leaflet heights and annular dimension (AML + PML - Ann) to be strikingly greater in the SAM group as compared with the No-SAM group (p = 0.001). CONCLUSION: A disparity between dimension of the annulus following mitral valve repair and combined heights of the two leaflets explains post-repair SAM. The AML height is a more important factor in the development of SAM. Thus, surgical techniques to reduce AML heights should be considered in patients with disproportionately large anterior leaflets in order to prevent SAM. Selection of size of the annuloplasty ring should take into consideration the height of the AML. PMID- 11380092 TI - The 'Pomeroy procedure': a new method to correct post-mitral valve repair systolic anterior motion. AB - Systolic anterior motion (SAM), a recognized complication of mitral valve repair, is often associated with left ventricular outflow gradient and mitral regurgitation. Current surgery to prevent these conditions is to perform sliding annuloplasty to reduce the posterior mitral leaflet (PML) height and to oversize the annuloplasty ring. However, these techniques do not consistently eliminate post-repair SAM, and removal of excess tissue and reduction of anterior mitral leaflet (AML) height may be more effective; this is the 'Pomeroy procedure'. Here, we report a patient in whom all standard procedures to prevent SAM were performed, but the condition still developed. This was corrected on a second pump run, using the Pomeroy procedure. PMID- 11380093 TI - The role of atrial contraction in mitral valve closure. AB - BACKGROUND AND AIM OF THE STUDY: Ovine mitral valve closure is associated with presystolic mitral annular reduction coincident with atrial contraction, which is abolished with ventricular pacing. Whether lack of properly timed atrial contraction influences mitral valve closure or competence, however, is not known. METHODS: Eight sheep underwent myocardial marker implantation on the left ventricle, mitral annulus (MA), and mitral leaflets. After 7-10 days, the animals were studied with biplane videofluoroscopy at baseline and during ventricular or atrioventricular (AV) sequential pacing. Valve closure was timed from end diastole (ED) and defined as minimum distance between two leaflet edge markers. ED was defined as peak of ECG R wave, end-systole as peak negative left ventricular (LV) dP/dt, and end-isovolumic contraction (EIVC) as 83.5 ms after ED. Septal-lateral (S-L) annular diameter was defined as distance between two markers at the middle of the anterior and posterior annulus. Regurgitant volume (RV) was calculated as relative volume change between ED and EIVC. RESULTS: V pacing was associated with delayed leaflet closure (65 +/- 5 versus 29 +/- 10 ms, p = 0.008); moreover, RV (4.1 +/- 0.5 versus 1.4 +/- 0.5 ml, p = 0.02), end diastolic S-L diameter (2.87 +/- 0.10 versus 2.67 +/- 0.09 cm, p = 0.0005), and MA area (8.12 +/- 0.37 versus 7.26 +/- 0.31 cm2, p = 0.009) all increased. RV and leaflet and annular dynamics during AV-pacing were similar to baseline. CONCLUSION: V-pacing increased S-L MA diameter by only 8 +/- 1%, but this change was associated with delayed leaflet coaptation and a 16 +/- 1% regurgitant fraction. These findings provide direct evidence that a properly timed atrial contraction is functionally important for effective mitral leaflet closure. PMID- 11380094 TI - Myxomatous mitral valve chordae. I: Mechanical properties. AB - BACKGROUND AND AIM OF THE STUDY: Chordal rupture is the most common reason for severe mitral regurgitation requiring surgery. The features that predispose myxomatous chordae to rupture, however, have not been studied. Thus, the physical and mechanical properties of normal and myxomatous mitral valve chordae were measured. METHODS: Chordae from 24 normal and 59 myxomatous mitral valves were cut into 10 mm-long segments and mechanically tested to measure extensibility, modulus, failure stress, failure strain, and failure load. After testing, the specimens were weighed and their cross-sectional area and volume measured. RESULTS: Chordae from myxoid mitral valves were larger (1.9 +/- 0.1 mm2 versus 0.8 +/- 0.1 mm2, p < or = 0.001) and heavier (16.6 +/- 1.0 mg versus 6.5 +/- 0.4 mg, p < or = 0.001) than normal chordae. Myxoid chordae had significantly lower moduli (40.4 +/- 10.2 MPa versus 132 +/- 15 MPa, p < or = 0.001) and failed at significantly lower tensile stress (6.0 +/- 0.6 MPa versus 25.7 +/- 1.8 MPa, p < or = 0.001) and absolute load (728 +/- 50 g versus 1,450 +/- 135 g, p < or = 0.001) than normal chordae. Normal and myxoid chordae had similar measurements of extensibility and failure strain. CONCLUSION: Myxomatous degeneration severely affects the mechanical properties of mitral valve chordae. Most notably, myxoid chordae failed at loads one-half of those of normal chordae. This may explain why chordal rupture is the main indication for repair of myxoid mitral valves. These findings also suggest that chordal preservation should be carried out with caution, as myxoid chordae are clearly abnormal with compromised mechanical strength. PMID- 11380095 TI - Myxomatous mitral valve chordae. II: Selective elevation of glycosaminoglycan content. AB - BACKGROUND AND AIM OF THE STUDY: Chordal rupture in myxomatous mitral valves is the leading cause of leaflet prolapse and regurgitation. Increased glycosaminoglycan (GAG) content has been reported in these valves. Therefore, the biochemical differences between myxomatous and control mitral valve chordae were investigated. METHODS: The contents of hexuronic acid, DNA, water, and collagen in chordae from 45 myxomatous valves and 10 control valves were measured. Collagen and hexuronic acid quantities were normalized to wet and dry weights, and to DNA content. Different GAG classes were measured using fluorophore assisted carbohydrate electrophoresis (FACE). RESULTS: Myxomatous chordae contained significantly more GAGs than controls after quantities were normalized for wet weight, dry weight, and DNA content. The FACE assay showed that the myxomatous chordae contained significantly more chondroitin/dermatan 6-sulfate when normalized to both wet and dry weight, and slightly more hyaluronan. In contrast to leaflets, which contain predominantly hyaluronan, the predominant GAG class in chordae was chondroitin/dermatan sulfate. Keratan sulfate, a GAG class previously unreported in valve tissues, was also discovered in the chordae. Myxomatous chordae contained more water and less collagen than control chordae, but equal quantities of DNA when normalized for wet weight. CONCLUSION: Cells in the chordae of myxomatous valves may produce more GAGs than cells in the chordae of control valves. The resulting accumulation of GAGs and bound water likely gives myxomatous valves their characteristic thickening and floppy, gelatinous nature, and may account for their reported mechanical weaknesses. PMID- 11380096 TI - The homograft aortic valve: a 29-year, 99.3% follow up of 1,022 valve replacements. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to elucidate the advantages and limitations of the homograft aortic valve for aortic valve replacement over a 29-year period. METHODS: Between December 1969 and December 1998, 1,022 patients (males 65%; median age 49 years; range: 1-80 years) received either a subcoronary (n = 635), an intraluminal cylinder (n = 35), or a full root replacement (n = 352). There was a unique result of a 99.3% complete follow up at the end of this 29-year experience. Between 1969 and 1975, homografts were antibiotic-sterilized and 4 degrees C stored (124 grafts); thereafter, all homografts were cryopreserved under a rigid protocol with only minor variations over the subsequent 23 years. Concomitant surgery (25%) was primarily coronary artery bypass grafting (CABG; n = 110) and mitral valve surgery (n = 55). The most common risk factor was acute (active) endocarditis (n = 92; 9%), and patients were in NYHA class II (n = 515), III (n = 256), IV (n = 112) or V (n = 7). RESULTS: The 30-day/hospital mortality was 3% overall, falling to 1.13 +/- 1.0% for the 352 homograft root replacements. Actuarial late survival at 25 years of the total cohort was 19 +/- 7%. Early endocarditis occurred in two of the 1,022 patient cohort, and freedom from late infection (34 patients) actuarially at 20 years was 89%. One-third of these patients were medically cured of their endocarditis. Preservation methods (4 degrees C or cryopreservation) and implantation techniques displayed no difference in the overall actuarial 20-year incidence of late survival endocarditis, thromboembolism or structural degeneration requiring operation. Thromboembolism occurred in 55 patients (35 permanent, 20 transient) with an actuarial 15-year freedom in the 861 patients having aortic valve replacement +/- CABG surgery of 92% and in the 105 patients having additional mitral valve surgery of 75% (p = 0.000). Freedom from reoperation from all causes was 50% at 20 years and was independent of valve preservation. Freedom from reoperation for structural deterioration was very patient age-dependent. For all cryopreserved valves, at 15 years, the freedom was 47% (0-20-year-old patients at operation), 85% (21-40 years), 81% (41-60 years) and 94% (>60 years). Root replacement versus subcoronary implantation reduced the technical causes for reoperation and re-replacement (p = 0.0098). CONCLUSION: This largest, longest and most complete follow up demonstrates the excellent advantages of the homograft aortic valve for the treatment of acute endocarditis and for use in the 20+ year-old patient. However, young patients (< or = 20 years) experienced only a 47% freedom from reoperation from structural degeneration at 10 years such that alternative valve devices are indicated in this age group. The overall position of the homograft in relationship to other devices is presented. PMID- 11380097 TI - A prospective study of changes in patients' quality of life after aortic valve replacement. AB - BACKGROUND AND AIM OF THE STUDY: After aortic valve replacement (AVR), 90% of survivors are in NYHA class I or II, and most return to work after three months. It is unclear, however, whether at that time patients have an improved quality of life (QOL), are able to perform physical and social activities, and live independently. METHODS: To assess this situation, 62 patients (39 males, 23 females; mean age 68.5 +/- 10 years) who underwent primary AVR completed the validated Short Form 36 (SF-36) questionnaire. QOL parameters were determined before and three months after surgery prospectively, and analyzed using the Wilcoxon matched pairs rank test. RESULTS: All patients showed significant improvement in all eight QOL parameters (score 0-100): (i) physical function (67 +/- 26 versus 37 +/- 28; p <0.00001); (ii) role limitation due to physical function (52 +/- 43 versus 20 +/- 37; p <0.0001); (iii) social function (80 +/- 25 versus 59 +/- 30; p <0.0001); (iv) role limitation due to emotional problems (64 +/- 41 versus 41 +/- 46; p = 0.01); (v) energy (62 +/- 22 versus 42 +/- 23; p <0.00001); (vi) mental health (78 +/- 19 versus 63 +/- 22; p <0.00001); (vii) pain (78 +/- 27 versus 67 +/- 32; p = 0.02); and (viii) general health perception (72 +/- 20 versus 58 +/- 21; p <0.00001). Significant improvements in QOL parameters were: (i) after mechanical and bioprosthetic AVR, seven of eight QOL parameters were improved; (ii) patients aged < 70 years had pronounced improvement in six of seven parameters, while those aged > or = 70 years had limited improvement in five of eight parameters; (iii) patients with valve sizes 19 and 21 mm improved in five of eight parameters, while those with > or = 23 mm implants improved in all eight parameters, regardless of post-AVR peak gradient of < 20 or > or = 20 mmHg; and (iv) patients with left ventricular end-diastolic dimension (LVEDD) < 55 mm improved in all eight parameters while those with LVEDD > or = 55 mm improved in only five parameters. CONCLUSION: There was significant improvement in patients' QOL at three months after AVR, regardless of the type of aortic implant used; improvement was greatest in those aged < 70 years. The results also suggest that patients with smaller implants (who were older) and those with LVEDD > or = 55 mm were less likely to show significant improvement in all QOL parameters at three months after AVR. PMID- 11380098 TI - Determinants of survival after aortic valve replacement as treatment for symptomatic aortic valve disease in the elderly. AB - BACKGROUND AND AIM OF THE STUDY: The effect of concurrent disease and cardiac comorbidity on survival after bioprosthetic valve replacement in elderly patients was assessed retrospectively. Risk factors were categorized as general, non cardiac (age, diabetes, previously treated carcinoma) and cardiac (LVEF, three vessel disease, previous CABG or valve replacement, and endocarditis). METHODS: A total of 400 elderly patients (median age 73 years; range: 71-76 years) was studied. Medical history included diabetes, previous CABG or aortic valve replacement (AVR), endocarditis and treatment of previous carcinoma. A left ventricular ejection fraction (LVEF) of <0.66 and presence of three-vessel disease were also investigated. Hospital deaths (and cause) were recorded; survival or date and cause of death after discharge were obtained by questionnaire. Kaplan-Meier univariate and Cox proportional hazards multivariate regression analyses were carried out. RESULTS: Mortality during follow up was 28.3%; hospital mortality was 3.8%. Univariate analysis showed five factors significantly to affect survival: LVEF, history of endocarditis, carcinoma, age and three-vessel disease. Fifteen of 38 patients with history of carcinoma died, 10 due to a malignancy. Of 76 patients with three-vessel disease, 26 died. A history of diabetes and previous CABG did not influence survival significantly. Four of eight patients with preoperative endocarditis died, all in hospital. Six of 11 patients died after redo-AVR, none before 36 months follow up. By Cox regression analysis, LVEF and histories of carcinoma and endocarditis remained significant. CONCLUSION: AVR should be performed before ventricular deterioration occurs. Previous CABG is not a contraindication for AVR. Endocarditis impaired survival. Long-term mortality after redo-AVR in this population was relatively high, but acceptable. AVR should also be performed in elderly patients with aortic valve disease. Since prognosis of symptomatic aortic valve disease is poor in the short term, AVR is indicated in patients treated for carcinoma. PMID- 11380099 TI - Electron beam computed tomography for the quantification of aortic valvular calcification. AB - BACKGROUND AND AIM OF THE STUDY: Calcific aortic stenosis is common in the elderly; indeed, 30-60% of patients with mild 'senile' aortic stenosis will progress to severe obstruction. Nonetheless, predictors of progression are incompletely defined, and non-invasive technologies capable of quantifying aortic valve calcium are needed. The reliability of electron beam computed tomography (EBCT) was evaluated for quantification of aortic valve calcium content. METHODS: Nineteen patients with and without restrictive valve calcification underwent EBCT scanning. Separate calcium scores, 30 s apart, were obtained in all patients, and the Spearman correlation coefficient was calculated between measurements. The relationship between dichotomized mean calcium score and aortic valve area was also investigated. RESULTS: There was excellent correlation between calcium scores (R = 0.99, p = 0.0001), as well as a significant inverse relationship between calcium scores in the upper and lower ranges and aortic valve area (p = 0.002). CONCLUSION: EBCT can be used for reproducible quantitation of aortic valve calcification. While at their extremes, calcium scores are inversely related to aortic valve area, further evaluation is needed to define the precise nature of this relationship throughout the spectrum of stenosis severity. EBCT holds promise in the longitudinal assessment of valvular calcification progression and its response to potential medical therapies. PMID- 11380100 TI - Surgical treatment of prosthetic valve endocarditis with left ventricular-aortic discontinuity: reconstruction of the left ventricular outflow tract with a xenopericardial conduit. AB - BACKGROUND AND AIM OF THE STUDY: Aortic prosthetic valve endocarditis (PVE) with annular destruction presents a challenge that requires techniques to eradicate the infection and correct the hemodynamic abnormality. METHODS: Between July 1, 1996 and March 31, 2000, six patients with native or PVE of the aortic valve and aortic annular destruction underwent surgical treatment. Of these patients, three (two men, one woman; mean age 71.0 years) had circumferential annular destruction of the aortic annulus, and formed the basis of this study. The microorganisms responsible for the infection were Streptococcus spp. in two patients and Staphylococcus aureus in one patient. In addition to aggressive debridement of the infected tissue, repair was achieved by reconstruction of the left ventricular outflow tract with a xenopericardial conduit and fixation of the new prosthetic valve to the conduit. RESULTS: One patient with ventricular septal perforation, multiple systemic embolism and sepsis died of low cardiac output syndrome soon after surgery. Two operative survivors were followed up for 9 and 51 months, with no late deaths. No patient has experienced recurrent infection, pericardial patch aneurysm, or prosthetic valve detachment. CONCLUSION: These operative procedures provide easy and secure fixation of the pericardial patch to the healthy tissue under excellent operative view, as well as a sturdy structure for the fixation of the new prosthesis, and complete exclusion of the abscess cavity from the blood stream. PMID- 11380101 TI - Experimental hypercholesterolemia induces apoptosis in the aortic valve. AB - BACKGROUND AND AIM OF THE STUDY: Aortic valve disease is presently the number one indication for valve replacement in the United States, yet its molecular mechanisms remain unknown. As apoptosis (programmed cell death) occurs in degenerative disease states, it was postulated that experimental hypercholesterolemia is associated with apoptosis in rabbit aortic valves. METHODS: New Zealand White rabbits (n = 8) were fed a 1% cholesterol diet for 12 weeks; control rabbits (n = 8) were fed a normal diet. After sacrifice of the animals, the aortic valves were dissected. Apoptosis was identified in the valvular lesion by TdT-mediated dUTP-biotin nick end-labeling (TUNEL) technique, and confirmed with transmission electron microscopy. The number of apoptotic cells was measured by computed morphometry. RESULTS: Valves from hypercholesterolemic rabbits showed an increase in apoptosis. TUNEL staining was identified in the atherosclerotic layer of hypercholesterolemic valves (0.1% of cells), but not in the cells of controls (p <0.0001). CONCLUSION: Apoptosis is increased in rabbit aortic valves during experimental hypercholesterolemia. If fatal cellular degeneration occurs in hypercholesterolemic valve disease, these data suggest that apoptosis may play a role in the mechanism of valvular disease. PMID- 11380102 TI - Aortic laceration due to prolapse of the bicuspid aortic valve: case report. AB - Reports of aortic regurgitation due to rupture of the aortic valve commissures are rare. Prompt surgical intervention is necessary, as the condition results in rapid, progressive heart failure and subsequent death. We report the case of a 78 year-old man who presented with aortic laceration and cardiac tamponade that was probably induced by prolapse of the bicuspid aortic valve. We speculate that prompt initial surgery may have prevented aortic laceration and cardiac tamponade in this patient. Thus, in order to optimize clinical outcome, clinicians must consider early, precautionary surgical management in patients who have sudden cardiac failure due to aortic regurgitation associated with prolapse of the bicuspid aortic valve. PMID- 11380103 TI - Aortic valve replacement in a patient with factor XII deficiency: case report. AB - Congenital factor XII deficiency is a rare condition. We report a case of aortic valve replacement (AVR) in a 63-year-old man with factor XII deficiency. On admission, the patient's activated partial thromboplastin time (aPTT) was prolonged (271 s), and activated clotting time was 500 s. His factor XII level was <3%. The Sonoclot signature showed an abnormal pattern. AVR with a prosthetic valve (St. Jude Medical) was performed safely after the normalization of aPTT and the Sonoclot signature by frozen plasma transfusion. The perioperative management in patients with factor XII deficiency is discussed. PMID- 11380104 TI - Anticoagulant management of patients with mechanical prosthetic valves undergoing non-cardiac surgery: indications and unresolved issues. AB - A wide array of recommendations is available for the management of anticoagulation in patients with a prosthetic heart valve scheduled for non cardiac surgery, ranging from avoidance of replacement anticoagulant therapy in all cases (excluding those with a recent thromboembolic event), to replacement anticoagulant therapy in all, without risk stratification. These guidelines are derived from only a few small- to medium-sized, non-randomized and often methodologically flawed studies conducted during the late 1970s, and applies mainly to caged-ball and caged-disc valves. Furthermore, extrapolation of the thromboembolic risk from data on patients not receiving oral anticoagulants at all is based on assumptions that are not necessarily valid. In this review, the direct and indirect evidence on which these guidelines are based is examined critically. Their applicability to the newer, less thrombogenic valve models is questionable. The need for further prospective, randomized studies is emphasized by the failure of existing studies to adjust properly for the main known or presumed thromboembolic risk factors, and their low statistical power to detect significant differences between protocols in an intention-to-treat manner. The evaluation of obstructive and non-obstructive thrombosis should serve as a secondary outcome measure in the assessment of anticoagulation management before non-cardiac surgery. PMID- 11380105 TI - Determination of plasma prothrombin level by Ca2+-dependent prothrombin activator (CA-1) during warfarin anticoagulation. AB - BACKGROUND AND AIM OF THE STUDY: The carinactivase-1 (CA-1) test is a new method for monitoring plasma prothrombin levels during warfarin anticoagulation therapy. METHODS: A total of 192 patients were allocated to two groups. Group A patients (n = 42) were controls (no warfarin); group B patients (n = 150) received warfarin. A Ca2+-ion and Boc-Val-Pro-Arg-pNA (a chromogenic substrate for thrombin) were added to 10-fold diluted plasma, after which prothrombin was activated with CA-1. Prothrombin levels were determined by measuring the extent of p-nitoroaniline liberation. RESULTS: The mean prothrombin level was 112.8 +/- 20.0 microg/ml in group A (Gaussian distribution), and 53.3 +/- 19.6 microg/ml in group B. In group B, correlations were found between the CA-1 test and prothrombin levels measured by prothrombin time (PT; r = 0.61, p <0. 01), PT-INR (r = 0.61, p <0.01), Thrombotest (TT; r = 0.57, p <0.01) and Hepaplastin test (HPT; r = 0.69, p <0.01). CONCLUSION: The CA-1 test represents a viable method of monitoring the coagulation system. CA-1 recognized the Gla-domain of prothrombin, and activated prothrombin. The CA-1 test required only 10 microl of diluted blood plasma, and took approximately 30 min to complete. The CA-1 test also measures prothrombin levels, correlates excellently with other tests for coagulation, and compares well with currently available methods for determining the efficacy of warfarin. PMID- 11380106 TI - Successful thrombolysis of prosthetic mitral valve thrombosis in early pregnancy. AB - Prosthetic valve thrombosis occurring during pregnancy is a life-threatening complication. Surgical treatment requires clot removal or valve replacement under cardiopulmonary bypass, and carries a high mortality. We report successful thrombolytic therapy with streptokinase for prosthetic valve thrombosis in a pregnant, 28-year-old woman. The patient, who had undergone mitral valve replacement (St. Jude Medical prosthesis) two years previously for restenosis after closed mitral valvotomy, was successfully thrombolyzed during the first trimester (6-8 weeks) for prosthetic valve thrombosis, and without any complication. The patient delivered a normal healthy child at nine months' gestation. Although thrombolysis in pregnancy has been reported previously, this is the first case in which it was performed during the first trimester for prosthetic valve thrombosis. PMID- 11380108 TI - Space-occupying lesions in the right ventricle of a patient with antiphospholipid syndrome. AB - A 29-year-old woman presented with shortness of breath, vague chest pain, and prominent intermittent ejection systolic murmur. Transthoracic echocardiography showed a large mass in the right ventricular outflow tract. Transesophageal echocardiography demonstrated two masses that were adherent to the tricuspid valve and intermittently prolapsed through the pulmonary valve. Computed tomography of the chest corroborated the echocardiographic findings. Currently, there are no definitive guidelines regarding the optimal management of right heart thrombi in patients with antiphospholipid syndrome. Our patient did not respond to a standard dose of rt-PA used in the treatment of pulmonary embolus. She underwent successful surgical resection of the thrombi without complications. PMID- 11380107 TI - Experience in corrective surgery for Ebstein's anomaly in 139 patients. AB - BACKGROUND AND AIM OF THE STUDY: The study aim was to summarize the authors' experience in corrective surgery of Ebstein's anomaly. METHODS: A total of 139 patients operated on between June 1980 and January 2000 was studied retrospectively. Among these patients, 111 underwent atrialized ventricle plication, tricuspid valve reconstruction and DeVega tricuspid annuloplasty, 27 underwent tricuspid valve replacement, and one patient with right ventricular hypoplasia underwent an additional total cavopulmonary connection. RESULTS: Overall, there were 12 operative deaths (mortality rate 8.6%); however, between 1990 and 2000, the mortality rate was 3.3%. Among the reconstruction patients, 10 cases were reoperated on for valve replacement, and all survived. CONCLUSION: Surgery for Ebstein's anomaly should be defined according to the pathologic/anatomic features of the condition. Tricuspid valve reconstruction should be performed in the mild condition; in medium A type, reconstruction should be performed, while for medium B type, reconstruction or valve replacement should be selected, albeit with caution. Valve replacement should be performed in the severe conditions. PMID- 11380109 TI - Comparative clinical outcomes with St. Jude Medical, Medtronic Hall and CarboMedics mechanical heart valves. AB - BACKGROUND AND AIM OF THE STUDY: Whether the St. Jude Medical (SJM), Medtronic Hall (MH) or CarboMedics (CM) heart valves confer any relative benefits to patient outcome remains controversial. While numerous studies have analyzed clinical results with a single brand, and a few studies have compared two brands, there are no single-center trials comparing all three valves. METHODS: Our experience with patients who had either a SJM, MH or CM mechanical valve in isolated aortic valve (AVR) or mitral valve (MVR) replacement was reviewed. AVR was performed in 953 patients (SJM = 394, MH = 314, CM = 245) and MVR in 591 patients (SJM = 193, MH = 264, CM = 134). Survivors were assessed annually; follow up consisted of 3336 patient-years (pt-yr) after AVR and 1693 pt-yr after MVR. RESULTS: Preoperatively, in the AVR group, more MH patients had previous valve surgery (p = 0.001) or were in NYHA class III/IV (p = 0.03), and more CM patients had a concomitant surgical procedure (p = 0.005). The hospital mortality after AVR with SJM, MH and CM valves was 3.8, 4.7 and 5.3%, respectively (p = 0.65). In the MVR group, there were more males in the CM group (p = 0.011), more CM patients had concomitant surgery (p = 0.001), and more MH patients had previous surgery (p = 0.006). The hospital mortality after MVR with SJM, MH and CM valves was 8.3, 10.2 and 6.0%, respectively (p = 0.35). There was no late survival advantage in either the AVR or MVR group according to the valve used (p = 0.24 and p = 0.90, respectively). For the AVR group the five-year actuarial freedom from thromboembolism was: SJM 85.8 +/- 2.5%, MH 80.1 +/- 2.7% and CM 85.9 +/- 3.5% (p = 0.04), and for MVR it was: SJM 84.2 +/- 4.0%, MH 77.5 +/- 3.4% and CM 86.9 +/- 5.2% (p = 0.27). Bleeding occurred with a similar frequency in the AVR (p = 0.36) and MVR (p = 0.70) groups. No cases of structural failure were identified in this study. At follow up, among AVR patients NYHA class III/IV was present in: SJM 5%, MH 6% and CM 3% (p = 0.50), while among MVR patients this was identified in: SJM 7%, MH 10% and CM 4% (p = 0.22). CONCLUSION: It is concluded that the SJM, MH and CM mechanical valves offer similar clinical results when used for isolated AVR or MVR. While there is a suggestion of an advantage with bileaflet valves, any differences detected may simply reflect differences in the preoperative patient variables. PMID- 11380110 TI - In vitro investigation of prosthetic heart valves in magnetic resonance imaging: evaluation of potential hazards. AB - BACKGROUND AND AIM OF THE STUDY: Magnetic resonance (MR) imaging is used in an increasing number of patients, and not only after cardiac valve replacement. However, ferromagnetic biomedical implants are often considered a contraindication for MR imaging because of the potential hazards with respect to their movement, dislodgement, or heating effects during the procedure. The purpose of this study was to assess ferromagnetism, attraction forces, heating effects, and artifacts associated with prosthetic heart valve implants. METHODS: Seventeen common heart valve prostheses (12 mechanical, five biological) were examined in vitro using a high-field-strength 1.5 Tesla (T) MR system. Attractive forces, temperature changes and the amount of artifacts were assessed by applying turbo-spin and gradient-echo sequences. RESULTS: The maximal calculated corresponding ferromagnetic force was (0.22 x 10(-3) N) in the static magnetic field. The temperature changes ranged from 0 to 0.5 degrees C maximum. Artifacts produced by the presence of the heart valve prostheses were less evident using a spin-echo sequence than a gradient-echo sequence. CONCLUSION: MR imaging exerted no significant force on the examined heart valve prostheses, and did not result in significant biological relevant temperature increase. None of the associated artifacts is considered to pose a substantial risk on MR imaging. MR procedures performed with a 1.5 T MR system can be applied safely in patients with heart valve prostheses evaluated in this study. PMID- 11380111 TI - Marfan's syndrome, dextrocardia and situs inversus associated with discrete subaortic stenosis and aortic insufficiency in an adult female: case report. AB - Marfan's syndrome is an inherited connective tissue defect that affects many organs, especially of the musculoskeletal, ophthalmic and cardiovascular systems, and may be associated with some rare conditions. Here, we report the first known case of Marfan's syndrome, combined with situs inversus totalis with dextrocardia and discrete subaortic stenosis and aortic insufficiency in a 22-year-old woman. PMID- 11380112 TI - Injuries to the elbow in childhood--a source of disquiet. PMID- 11380113 TI - The treatment of metastases in the appendicular skeleton. PMID- 11380114 TI - How should orthopaedic surgeons respond to unsolicited e-mail? PMID- 11380115 TI - Instability due to unrecognised fracture-subluxations after apparently isolated injuries of the cervical spine. AB - Instability may present at a different level after successful stabilisation of an unstable segment in apparently isolated injuries of the cervical spine. It can give rise to progressive deformity or symptoms which require further treatment. We performed one or more operations for unstable cervical spinal injuries on 121 patients over a period of 90 months. Of these, five were identified as having instability due to an initially unrecognised fracture-subluxation at a different level. We present the details of these five patients and discuss the problems associated with their diagnosis and treatment. PMID- 11380116 TI - The association of degeneration of the intervertebral disc with 5a/6a polymorphism in the promoter of the human matrix metalloproteinase-3 gene. AB - It has been suggested that matrix metalloproteinase-3 (MMP-3, stromelysin-1) has an important role in the degeneration of intervertebral discs (IVDs). A human MMP 3 promoter 5A/6A polymorphism was reported to be involved in the regulation of MMP-3 gene expression. We suggest that IVD degeneration is associated with 5A/6A polymorphism. We studied 54 young and 49 elderly Japanese subjects. Degeneration of the lumbar discs was graded using MRI in the younger group and by radiography in the elderly. 5A/6A polymorphism was determined by polymerase-chain reaction based assays. We found that the 5A5A and 5A6A genotype in the elderly was associated with a significantly larger number of degenerative IVDs than the 6A6A (p < 0.05), but there was no significant difference in the young. In the elderly, the IVD degenerative scores were also distributed more highly in the 5A5A and 5A6A genotypes (p = 0.0029). Our findings indicate that the 5A allele is a possible risk factor for the acceleration of degenerative changes in the lumbar disc in the elderly. PMID- 11380117 TI - Excision of hemivertebrae in the management of congenital scoliosis involving the thoracic and thoracolumbar spine. AB - We present a study of ten consecutive patients who underwent excision of thoracic or thoracolumbar hemivertebrae for either angular deformity in the coronal plane, or both coronal and sagittal deformity. Vertebral excision was carried out anteriorly alone in two patients. Seven patients had undergone previous posterior spinal fusion. Their mean age at surgery was 13.4 years (6 to 19). The mean follow-up was 78.5 months (20 to 180). The results were evaluated by radiological review of the preoperative, postoperative and most recent follow-up films. The mean preoperative coronal curve was 78.2 degrees (30 to 115) and was corrected to 33.9 degrees (7 to 58) postoperatively, a mean correction of 59%. Preoperative coronal decompensation of 35 mm was improved to 11 mm postoperatively. Seven patients had significant coronal decompensation preoperatively, which was corrected to a physiological range postoperatively. There were no major complications and no neurological damage. We have shown that resection of thoracic and thoracolumbar hemivertebrae can be performed safely, without undue risk of neurological compromise, in experienced hands. PMID- 11380118 TI - The risks and benefits of percutaneous nucleotomy for lumbar disc herniation. A 10-year longitudinal study. AB - Percutaneous nucleotomy is a relatively new technique for treating lumbar disc herniation. There is no agreement as to the volume of disc material to be removed. A long-term study of clinical and radiological data from patients treated by percutaneous nucleotomy was designed to identify the factors associated with favourable and unfavourable outcomes. We studied 42 patients for at least ten years; the mean follow-up was 10.9 years. They were divided into two subgroups to assess the value of preserving the nucleus pulposus in the central area of the disc. The overall success rate for both subgroups was 50%. A decrease in disc height on plain radiography and a decrease in signal intensity on MRI were observed more infrequently in patients in whom the nucleus pulposus in the central area of the disc had been preserved, than in those in whom it had been extensively removed. These adverse radiological findings correlated closely with increased low back pain during the first one to two years after operation and a poorer overall outcome. We conclude that percutaneous nucleotomy is most likely to be successful when the central area of the disc is preserved. PMID- 11380119 TI - Factors affecting employment after whiplash injury. AB - Of 586 employed patients with a whiplash injury 40 (7%) did not return to work. The risk was increased by three times in heavy manual workers, two and a half times in patients with prior psychological symptoms and doubled for each increase of grade of disability. The length of time off work doubled in patients with a psychological history and trebled for each increase in grade of disability. The self-employed were half as likely to take time off work, but recovered significantly more slowly than employees. PMID- 11380120 TI - Long-term results of retention of the posterior cruciate ligament in total knee replacement in rheumatoid arthritis. AB - We analysed the long-term results with a mean follow-up of 10.2 years, of 66 total knee replacements in 42 patients with rheumatoid arthritis. In all cases the posterior cruciate ligament was retained. There were only three complications (4.5%). Revision surgery was necessary in five knees (7.6%), including one (1.5%) with infection. At the final follow-up, 75.8% of knees were rated excellent clinically. Only 15% had an excellent function score. The survival rate of the implant was 90.7% at 19 years. The clinical, radiological and survivorship analysis shows that the posterior-cruciate-retaining knee arthroplasty performs well in rheumatoid arthritis. PMID- 11380121 TI - Long-term results of arthroscopic partial lateral meniscectomy in knees without associated damage. AB - We evaluated the outcome of partial lateral meniscectomy of 31 knees in 29 patients whose knees were otherwise normal. The mean follow-up was 10.3 years. According to the Lysholm score, 14 knees were rated as excellent, four as good, five as fair and eight as poor, with a mean score of 80.5 points. Radiologically, only one lateral compartment was classified as grade 0, eight as grade 1, nine as grade 2, 11 as grade 3, and two as grade 4 according to Tapper and Hoover. No significant (p < 0.05) correlation was found between the amount of tissue resected and the subjective, clinical and radiological outcome. Although early results of lateral meniscectomy may be satisfactory, we have demonstrated that in the long term there was a high incidence of degenerative changes, a high rate of reoperation (29%) and a relatively low functional outcome score. PMID- 11380122 TI - Penetrating missile injuries of the brachial plexus. AB - We studied a consecutive series of 58 patients with penetrating missile injuries of the brachial plexus to establish the indications for exploration and review the results of operation. At a mean of 17 weeks after the initial injury, 51 patients were operated on for known or suspected vascular injury (16), severe persistent pain (35) or complete loss of function in the distribution of one or more elements of the brachial plexus (51). Repair of the nerve and vascular lesions abolished, or significantly relieved, severe pain in 33 patients (94%). Of the 36 patients who underwent nerve graft of one or more elements of the plexus, good or useful results were obtained in 26 (72%). Poor results were observed after repairs of the medial cord and ulnar nerve, and in patients with associated injury of the spinal cord. Neurolysis of lesions in continuity produced good or useful results in 21 of 23 patients (91%). We consider that a vigorous approach is justified in the treatment of penetrating missile injury of the brachial plexus. Primary intervention is mandatory when there is evidence of a vascular lesion. Worthwhile results can be achieved with early secondary intervention in patients with debilitating pain, failure to progress and progression of the lesion while under observation. There is cause for optimism in nerve repair, particularly of the roots C5, C6 and C7 and of the lateral and posterior cords, but the prognosis for complete lesions of the plexus associated with damage to the cervical spinal cord is particularly poor. PMID- 11380123 TI - Conservative versus operative treatment for displaced ankle fractures in patients over 55 years of age. A prospective, randomised study. AB - Forty-seven patients over the age of 55 years with a displaced fracture of the ankle were entered into a prospective, randomised study in order to compare open reduction and internal fixation with closed treatment in a plaster cast; 36 were reviewed after a mean of 27 months. The outcome was assessed clinically, radiologically and functionally using the Olerud score. The results showed that anatomical reduction was significantly less reliable (p = 0.03) and loss of reduction significantly more common (p = 0.001) in the group with closed treatment. Those managed by open reduction and internal fixation had a significantly higher functional outcome score (p = 0.03) and a significantly better range of movement of the ankle (p = 0.044) at review. PMID- 11380124 TI - Dislocation of the knee with lateral dislocation of the patella. A report of four cases. AB - In a group of 25 patients with traumatic dislocation of the knee, four, all of whom had similar ligament and medial soft-tissue injuries, also had associated lateral patellar dislocation. In all four reconstruction was delayed because of their other serious injuries. Having encountered the combination of knee dislocation and lateral patellar dislocation in 16% of our patients, we believe that it may be less rare than is commonly believed. We think that it is important to maintain a high index of suspicion of possible patellar dislocation when medial structures have been severely damaged. Early recognition and immobilisation in extension can prevent fixed lateral dislocation of the patella. PMID- 11380125 TI - Fracture stiffness as a guide to the management of tibial fractures. AB - We have studied the progression of healing in 103 unstable fractures of the tibia. In 76 patients we removed the external fixator once the stiffness had reached 15 Nm/degrees in the sagittal plane. Deformity at the site of the fracture subsequently occurred in four patients. In a further 27, we measured stiffness in several planes and removed the fixator only when the stiffness reached 15 Nm/degrees in each. We found that stiffness in two orthogonal planes may differ widely (maximum difference 9.0 Nm/degrees, mean 4.1 Nm/degrees). There were no failures in the second group. We advocate that fracture stiffness be measured in two orthogonal planes when assessing tibial healing and suggest that values above 15 Nm/degrees in two planes give an indication that it is safe to remove the fixator. PMID- 11380126 TI - Fractures of the neck of the radius in children. Early experience with intramedullary pinning. AB - The treatment of fractures of the neck of the radius in children is difficult, particularly if the angulation of the fracture exceeds 60 degrees. Since 1994 we have used closed reduction and stabilisation with an intramedullary Kirschner wire in patients with grade-IV fractures according to the classification of Judet et al. In a retrospective analysis of a two-year period (1994 to 1996), 324 children with fractures of the elbow were treated in our department. Of these, 29 (9%) had a fracture of the neck of the radius; six were grade-IV injuries (1.9%). Five of the latter had an excellent postoperative result with normal movement of the elbow and forearm. One patient with a poor result had a concomitant dislocation of the elbow. Our results suggest that closed reduction and intramedullary pinning of grade-IV fractures allows adequate stabilisation while healing occurs. PMID- 11380127 TI - Arthrography for reduction of a fracture of the radial neck in a child with a non ossified radial epiphysis. AB - A fracture of the neck of the radius when the head is not ossified can be difficult to assess and treat. In a four-year-old child we suspected from the radiographs that there was an O'Brien type-III injury after trauma. Partial manual reduction of the non-ossified radial head was completed using the Metaizeau technique of intramedullary Kirschner (K-) wiring aided by intraoperative arthrography. The child had a full range of movement at the elbow and wrist when reviewed 11 weeks after the injury, three weeks after removal of the K-wire. We suggest that intraoperative arthrography is a useful complement to the Metaizeau technique for successful reduction of fractures of the radial neck in the presence of a non-ossified radial head. PMID- 11380128 TI - The value of MRI in the assessment of an elbow injury in a neonate. AB - We describe the use of MRI to establish the exact diagnosis in a swollen elbow in a neonate. Urgent diagnosis was needed for medical and social reasons. We accomplished this without the use of an invasive procedure or anaesthesia for a fracture that is recognised to be difficult to diagnose in patients of this age group. PMID- 11380129 TI - Missed chronic anterior Monteggia lesion. Closed reduction by gradual lengthening and angulation of the ulna. AB - Two consecutive cases of chronic dislocation of the head of the radius after missed Bado type-I Monteggia lesions are presented. Reduction was successfully achieved in both patients after ulnar corticotomy, gradual lengthening and angulation of the ulna using an external fixator. Open reduction or reconstruction of the radio-ulnar capitellar joint was not undertaken. The age at injury was seven years in the older and two years in the younger patient. The time from injury to treatment was five years in the older and three months in the younger child. At follow-up, nine years after completion of treatment in the older and eight months in the younger patient, both show satisfactory movement, function of the forearm and reduction of the head of the radius. This technique may be considered in missed Monteggia lesions before open procedures on the radio ulnar capitellar joint are undertaken. PMID- 11380130 TI - Deformities of the shoulder in infants younger than 12 months with an obstetric lesion of the brachial plexus. AB - We performed a prospective study using MRI in 16 consecutive infants with a mean age of 5.2 months (2.7 to 8.7) who had shown inadequate recovery from an obstetric lesion of the brachial plexus in the first three months of life, in order to identify early secondary deformities of the shoulder. Shoulders were analysed according to a standardised MRI protocol. Measurements were made of the appearance of the glenoid, glenoid version and the position of the humeral head. The appearance of the glenoid on the affected side was normal in only seven shoulders. In the remainder it was convex in seven and bioconcave in three. The degree of subluxation of the humeral head was significantly greater (p = 0.01) in the affected shoulders than in normal shoulders (157 degrees v 170 degrees). The presence of an abnormal appearance of the glenoid, retroversion of the glenoid and subluxation of the humeral head increased with age. There was a statistical difference (p = 0.05) between infants younger than five months and those who were older. PMID- 11380131 TI - Hospital versus home management of children with buckle fractures of the distal radius. A prospective, randomised trial. AB - Our aim was to determine whether children with buckle fractures of the distal radius could be managed at home after initial hospital treatment. There were 87 patients in the trial: 40 had their short-arm backslab removed at home three weeks after the initial injury, and 47 followed normal practice by attending the fracture clinic after three weeks for removal of the backslab. Clinical examination six weeks after the injury showed no significant difference in deformity of the wrist, tenderness, range of movement and satisfaction between the two groups. Fourteen (33%) of the hospital group compared with five (14%) (p = 0.04) of those managed in the community stated that they had problems with the care of their child's fracture. It was found that both groups, given a choice, would prefer to remove their child's backslab at home (p < 0.001). Our findings show that it is clinically safe to manage children with buckle fractures within the community. PMID- 11380132 TI - Localised deposition of amyloid in tears of the rotator cuff. AB - Age-related localised deposition of amyloid in connective tissue has been found in degenerative articular and periarticular tissue. Biopsies of the supraspinatus tendon of 28 patients undergoing repair of the rotator cuff were analysed histologically for the presence of localised deposition of amyloid. There was a long history of impingement in 20 patients, and eight patients had suffered an acute traumatic tear with no preceding symptoms. Localised deposition of amyloid identified by Congo Red staining was detected in 16 samples (57%). Amyloid was present in 14 (70%) of the degenerative tears, but in only two (25%) of the acute tears. Immunohistochemical staining showed that the amyloid deposits were positive for P component, but negative for kappa and lambda light chains, prealbumin, and beta2 microglobulin. Critical electrolyte staining revealed highly-sulphated glycosaminoglycans at sites of deposition of amyloid. The presence of localised deposition of amyloid in tears of the rotator cuff is likely to represent irreversible structural changes. These findings support the theory that impingement and tears are due to intrinsic degenerative changes within the tendons of the rotator cuff. PMID- 11380133 TI - The cross-sectional area of supraspinatus as measured by diagnostic ultrasound. AB - We have established a reference standard for the cross-sectional area (CSA) of supraspinatus as measured by diagnostic ultrasound. The influence of hand dominance and of ageing on the CSA was also assessed. We examined 72 subjects aged from 20 to 79 years. Standard values of the CSA were determined with a high measure of interobserver reliability. Although the CSA on the dominant side was significantly larger (p < 0.001) by 0.16 cm2 (95% CI 0.072 to 0.249) than that on the non-dominant side, this difference had no clinical significance. The CSA of supraspinatus decreased significantly with ageing. PMID- 11380134 TI - Perthes' disease. A study of constitutional aspects in adulthood. AB - We studied 16 patients suffering from osteoarthritis of the hip who had had Perthes' disease during childhood. They were compared clinically and radiologically with a control group who had not had Perthes' disease, in order to assess whether a generalised, pre-existing constitutional disorder was present. Nine patients with a previous history of Perthes' disease had some other skeletal abnormality, but only three presented with clinical symptoms. Only one patient in the control group was found to have an abnormality but was symptom-free. Our findings provide further evidence that patients with Perthes' disease may have a generalised abnormality related to chondrogenesis which can produce other skeletal anomalies that persist into adult life. PMID- 11380135 TI - Femoral shortening in intertrochanteric fractures. A comparison between the Medoff sliding plate and the compression hip screw. AB - We compared 54 patients treated by a Medoff sliding plate (MSP) with 60 stabilised by a compression hip screw (CHS) in a prospective, randomised study of the management of intertrochanteric femoral fractures. Four months after the operation femoral shortening was determined from radiographs of both femora. In unstable fractures the mean femoral shortening was 15 mm with the MSP and 11 mm with the CHS (p = 0.03). A subgroup in which shortening was classified as large, comprising one-third of the patients in each group, had a similar extent of shortening, but more medialisation of the femoral shaft occurred in the CHS (26%) than in the MSP (12%) group (p = 0.03). Five postoperative failures of fixation occurred with the CHS and none with the MSP (p = 0.03). The marginally greater femoral shortening seen with the MSP compared with the CHS appeared to be justified by the improved control of impaction of the fracture. Biaxial dynamisation in unstable intertrochanteric fractures is a safe principle of treatment, which minimises the rate of postoperative failure of fixation. PMID- 11380136 TI - Hip disease and the prognosis of total hip replacements. A review of 53,698 primary total hip replacements reported to the Norwegian Arthroplasty Register 1987-99. AB - We studied the rates of revision for 53,698 primary total hip replacements (THRs) in nine different groups of disease. Factors which have previously been shown to be associated with increased risk of revision, such as male gender, young age, or certain types of uncemented prosthesis, showed important differences between the diagnostic groups. Without adjustment for these factors we observed an increased risk of revision in patients with paediatric hip diseases and in a small heterogeneous 'other' group, compared with patients with primary osteoarthritis. Most differences were reduced or disappeared when an adjustment for the prognostic factors was made. After adjustment, an increased relative risk (RR) of revision compared with primary osteoarthritis was seen in hips with complications after fracture of the femoral neck (RR = 1.3, p = 0.0005), in hips with congenital dislocation (RR = 1.3, p = 0.03), and in the heterogenous 'other' group. The analyses were also undertaken in a more homogenous subgroup of 16,217 patients which had a Charnley prosthesis implanted with high-viscosity cement. The only difference in this group was an increased risk for revision in patients who had undergone THR for complications after fracture of the femoral neck (RR = 1.5, p = 0.0005). THR for diagnoses seen mainly among young patients had a good prognosis, but they had more often received inferior uncemented implants. If a cemented Charnley prosthesis is used, the type of disease leading to THR seems in most cases to have only a minor influence on the survival of the prosthesis. PMID- 11380137 TI - The effects of particulate bone cements at the bone-implant interface. AB - We used a rat model in vivo to study the effects of particulate bone cements at the bone-implant interface. A ceramic pin was implanted into the tibiae of 48 rats. Three types of particle of clinically relevant size were produced from one bone-cement base without radio-opacifier, with zirconium dioxide (ZrO2) and with barium sulphate (BaSO4). The rats were randomly assigned to four groups to receive one of the three bone cements or normal saline with 2% v/v Sprague-Dawley serum as the control. A total of 10(9) particles was injected into the knee at 8, 10 and 12 weeks after the original surgery. The animals were killed at 14 weeks and the tibiae processed for histomorphometry. The area of fibrous tissue and the gap between the implant and bone were measured using image analysis. All three types of particle were associated with a larger area of bone resorption than the control. Only in the case of the BaSO4-containing cement did this reach statistical significance (p = 0.01). Particles of bone cement appear to promote osteolysis at the bone-implant interface and this effect is most marked when BaSO4 is used as the radiopaque agent. PMID- 11380138 TI - Differential effects of oxidised and non-oxidised polyethylene particles on human monocyte/macrophages in vitro. AB - Sterilisation by gamma irradiation in the presence of air causes free radicals generated in polyethylene (PE) to react with oxygen, which could lead to loss of physical properties and reduction in fatigue strength. Tissue retrieved from failed total hip replacements often has large quantities of particulate PE and most particles associated with peri-implant osteolysis are oxidised. Consequently, an understanding of the cellular responses of oxidised PE particles may lead to clarification of the pathogenesis of osteolysis and aseptic loosening. We have used the agarose system to demonstrate the differential effects of oxidised and non-oxidised PE particles on the release of proinflammatory products such as interleukin-1beta (IL-1beta), IL-6, and tumour necrosis factor-alpha (TNF-alpha) from monocytes/macrophages (M/M). Oxidised PE particles were shown to stimulate human M/M to phagocytose and to release cytokines. Oxidation may alter the surface chemistry of the particles and enhance the response to specific membrane receptors on macrophages, such as scavenger type receptors. PMID- 11380139 TI - Comparative analysis of two different types of alumina-alumina hip prosthesis retrieved for aseptic loosening. AB - We compared and quantified the modes of failure and patterns of wear of 11 Mittelmeier and 11 Ceraver-Ostal retrieved alumina-alumina hip prostheses with reference to the corresponding clinical and radiological histories. Macroscopic wear was assessed using a three-dimensional co-ordinate measuring machine. Talysurf contacting profilometry was used to measure surface roughness on a microscopic scale and SEM to determine mechanisms of wear at the submicron level. The components were classified into one of three categories of wear: low (no visible/measurable wear), stripe (elliptical wear stripe on the heads and larger worn areas on the cups) and severe (macroscopic wear, large volumes of material lost). Overall, the volumetric wear of the alumina-alumina prostheses was substantially less than the widely used metal and ceramic-on-polyethylene combinations. By identifying and eliminating the factors which accelerate wear, it is expected that the lifetime of these devices can be further increased. PMID- 11380140 TI - Recurring synovitis as a possible reason for aseptic loosening of knee endoprostheses in patients with rheumatoid arthritis. AB - We evaluated histologically samples of synovial tissue from the knees of 50 patients with rheumatoid arthritis (RA). The samples were taken during revision for aseptic loosening. The findings were compared with those in 64 knees with osteoarthritis (OA) and aseptic loosening and in 18 knees with RA without loosening. The last group had been revised because of failure of the inlay or the coupling system of a constrained prosthesis. All the patients had had a total ventral synovectomy before implantation of the primary prosthesis. In all three groups a foreign-body reaction and lymphocellular infiltration were seen in more than 80% of the tissue samples. Deposits of fibrin were observed in about one third to one-half of the knees in all groups. Typical signs of the reactivation of RA such as rheumatoid necrosis and/or proliferation of synovial stromal cells were found in 26% of knees with RA and loosening, but not in those with OA and loosening and in those with RA without loosening. Our findings show that reactivation of rheumatoid synovitis occurs after total knee replacement and may be a cofactor in aseptic loosening in patients with RA. PMID- 11380141 TI - Combined anteroposterior spinal fixation provides superior stabilisation to a single anterior or posterior procedure. AB - Fusion is the main goal in the surgical management of the injured and unstable spine. A wide variety of implants is available to enhance this. Our study was performed to evaluate the stabilising characteristics of several anterior, posterior and combined systems of fixation. Six thoracolumbar (T11 to L2) spines from 13-week-old calves were first tested intact. Then the vertebral body of T13 was removed and the defect replaced and supported by a wooden block to simulate bone grafting. Dorsal implants consisting of a Universal Spine System (USS) fracture system and an AO Fixateur interne (AOFI), and ventral implants comprising of a Kaneda Classic, a Kaneda SR, a prototype of the VentroFix single clamp/single rod construct (SC/SR) and the VentroFix single clamp/double rod construct (SC/DR) were first implanted individually to stabilise the removal of the vertebral body. Simulating the combined anteroposterior stabilisations, all ventral implants were combined with the AOFI. The range of motion (ROM) was measured under loads of up to 7.5 Nm. The load was applied in a custom-made spine tester in the three primary directions while measuring the intervertebral movements using a goniometric linkage system. The dorsal systems limited ROM in flexion below 0.9 degrees and in extension between 3.3 degrees and 3.6 degrees (median values). The improved Kaneda System SR yielded a mean ROM of 1.8 degrees in flexion and in extension. The median rotation found with the VentroFix (SC/DR) was 3.2 degrees for flexion and 2.8 degrees for extension. Reinforcement of the ventral constructs with a dorsal system reduced the ROM in flexion and extension in all cases to 0.4 degrees and lower. In rotation, the median ROM of the anterior systems ranged from 2.7 degrees to 5.1 degrees and for the posterior systems from 3.9 degrees to 5.7 degrees, while the combinations provided a ROM of 1.2 degrees to 1.9 degrees. In lateral bending, the posterior implants restricted movement to 1.1 degrees, whereas the anterior implants allowed up to 5.2 degrees. The combined systems provided the highest stability at less than 0.6 degrees. Our study revealed distinct differences between posterior and anterior approaches in all primary directions. Also, different stabilisation characteristics were found within the anterior and posterior groups. Combinations of these two approaches provided the highest stability in all directions. PMID- 11380142 TI - The management of intracapsular fractures of the proximal femur. PMID- 11380143 TI - The management of intracapsular fractures of the proximal femur. PMID- 11380144 TI - The teaching of trauma and orthopaedic surgery to the undergraduate in the United Kingdom. PMID- 11380145 TI - Pyomyositis of the iliacus muscle in a child. PMID- 11380146 TI - Does 'canal clearance' affect neurological outcome after thoracolumbar burst fractures? PMID- 11380147 TI - Spontaneous bifocal Clostridium septicum gas gangrene. PMID- 11380148 TI - Non-M protein(s) on the cell wall and membrane of group A streptococci induce(s) IFN-gamma production by dermal CD4+ T cells in psoriasis. AB - Recently we have demonstrated that a disease-specific subpopulation of CD4+ T cells isolated from skin lesions of chronic plaque psoriasis produces interferon gamma in response to group A streptococcal (GAS) antigens. To determine if these T cells recognize M or non-M protein, extracts from cell wall of type M6 GAS (M6W) and its isogenic M gene deletion mutant (M-W), M6 membrane extract (M6M) and recombinant M6 protein (rM6) were used to stimulate GAS-reactive T-cell lines from nine patients with chronic plaque psoriasis. T-cell lines were incubated with or without streptococcal extracts for 18 h in the presence of a transport inhibitor, stained for surface CD4 and intracellular cytokine expression, and analysed by flow cytometry. Variable numbers (0.2-34%) of CD4+ T cells produced interferon-gamma, in all but one of the T-cell lines tested, in response to M6W, M-W and M6M extracts. No significant difference between the response to M6W and M W extracts was detected. In addition, rM6 protein failed to increase CD4+/interferon-gamma+ T-cell numbers in seven of nine T-cell lines compared to medium alone. For the group, there was a highly significant correlation between the responses to the three extracts (M6W vs M-W, P = 0.0005; M6W vs M6M, P = 0.0003; M-W vs M6M, P = 0.0001). Low or minimal numbers of interleukin-4- and interleukin-10-producing CD4+ T cells were occasionally induced. These findings suggest that a subpopulation of CD4+ T cells isolated from skin lesions of chronic plaque psoriasis patients produces interferon-gamma in response to non-M protein(s) present on the cell wall and membrane of GAS. PMID- 11380149 TI - Upregulation of histidine decarboxylase mRNA expression in scleroderma skin. AB - The involvement of histamine in the pathogenesis of systemic sclerosis (SSc) has been suggested. Possible involvement of histamine and histidine decarboxylase (HDC), the synthesizing enzyme for histamine, in the formation of the skin abnormalities in SSc was studied. Skin histamine concentrations in SSc were significantly lower than in normal controls (P < 0.02). In situ hybridization with an HDC probe revealed that the expression of the HDC gene in SSc was greater than in normal controls. The number of cells and the mean grain number per cell expressing HDC mRNA were both significantly greater in SSc than in normal controls (both P < 0.01). These results show a reduction in histamine concentration and an elevated HDC gene expression in SSc skin, indicating an increase in both histamine release and HDC gene expression. The upregulation of histamine turnover appears to be involved in the formation of the skin abnormalities of SSc. PMID- 11380150 TI - Opposing effects of UVA1 phototherapy on the expression of bcl-2 and p53 in atopic dermatitis. AB - Recently, medium-dose UVA1 phototherapy (50 J/cm2) has been introduced as an effective treatment for severe atopic dermatitis (AD). In order to further elucidate the mechanisms by which medium-dose UVA1 irradiation leads to an improvement in skin status in patients with AD, biopsy specimens from ten patients before and after treatment were analysed immunohistochemically for features of apoptosis. We sought to determine the extent to which UVA1 irradiation was able to modulate the balance between p53 and bcl-2 expression in vivo using monoclonal antibodies labelling these proteins. As compared with lesional skin of patients with AD before UVA1 irradiation, the number of dermal cells, apparently lymphocytes, that were positive for p53 had significantly increased after treatment and, in addition, some basal keratinocytes showed slight positive staining for p53. An increased expression of the bcl-2 gene before treatment in predominately dermal lymphocytes was significantly downregulated by UVA1 therapy. The increase in p53+ cells and the decrease in bcl 2+ cells were closely linked to a significant reduction in dermal T cells (CD3+) and a substantial clinical improvement in skin condition. In summary, medium-dose UVA1 irradiation led to a marked modulation of the expression of p53 and bcl-2, and this plays a key role in regulating UVA1-induced apoptosis. PMID- 11380151 TI - A modified ex vivo skin organ culture system for functional studies. AB - To investigate the immunological function of cells in normal and diseased skin under conditions approximating the in vivo situation, it is necessary to maintain the structural integrity of the tissue. To achieve this, freshly isolated skin has to be cultured ex vivo, or an in vitro-constructed complete skin equivalent may be used. Different skin organ culture systems have been described. Basically two systems prevail: submerged or air-exposed skin organ cultures. The former model has been used for measuring cytokine secretion by skin cells in the medium, and the latter to study the expression of cell membrane proteins in situ and the kinetics of epidermal Langerhans cells. Here we present a modified ex vivo skin organ culture system which approaches the in vivo situation by maintaining the normal skin architecture without spontaneous induction of the regenerative maturation markers. This method allowed the expression of cell membrane proteins in situ to be measured, and the cytokine secretion by skin cells in the culture medium to be quantitated in the same experiment. In this system, both general and specific stimuli (LPS and IL-1beta) upregulated the expression of skin-derived cytokines IL-8 and IL-6 in the medium and different markers (ICAM-1, CD40 and CD86) on cells in the tissue in a 24-hour culture-formed. Elevation of both cytokine and cell marker expression could be blocked by dexamethasone and by IL 1ra which acts specifically on IL-1beta-mediated responses. The system presented here is both quick and simple and can be used as a model to study the behaviour of skin cells in their natural microenvironment. PMID- 11380152 TI - Determination of stratum corneum lipid profile by tape stripping in combination with high-performance thin-layer chromatography. AB - Intercellular lipids in the stratum corneum (SC) are responsible for the barrier function of mammalian skin. The main components of the SC lipids are ceramides, cholesterol, and free fatty acids, as established by thin-layer chromatographic analysis of lipids extracted from the human and mammalian SC. Up to now, for lipid analysis the extracts of the entire SC has been used and information on whether the lipid composition changes with the depth in the SC is scarce. Tape stripping is a technique which removes corneocyte layers step by step with an adhesive film. The use of this technique for lipid analysis was hampered by the contamination of lipid extracts with compounds co-extracted from the tape with organic solvents used for the extraction of SC lipids. The aim of the present study was to establish a suitable analytical method for the determination of the local SC lipid composition. For this purpose, the SC samples were collected by sequential stripping with Leukoplex tape in five healthy volunteers. The lipids were extracted with ethyl acetate:methanol mixture (20:80) and separated by means of HPTLC. The results of this study revealed that the free fatty acid level is highest and the cholesterol and ceramide levels lowest in the uppermost SC layers (about 4 strippings). The levels remained unchanged in the underlying SC layers. In these layers, the ceramide level was about 60 wt% and the free fatty acid and cholesterol levels were about 20 wt% each. Ceramides could be separated into seven different fractions and the relative amounts of individual ceramide fractions did not significantly change with the SC depth. Cholesterol sulfate levels were about 5% of total cholesterol and did not change with the SC depth, except for the for the first strip where the level was about 1%. The method developed makes it possible to study the differences in the SC lipid profile in healthy and diseased human skin with relation to the SC lipid organization and to the skin barrier function in vivo. PMID- 11380153 TI - Growth suppression of hamster flank organs by topical application of catechins, alizarin, curcumin, and myristoleic acid. AB - Hamster flank organ growth, as measured by an increase in the area of the pigmented macule, is androgen-dependent. When flank organs of a castrated hamster are treated topically with testosterone, the flank organ becomes larger and darker. Since this growth is known to be dependent on the intracellular active androgen, 5alpha-dihydrotestosterone (DHT), inhibitors of 5alpha-reductase which converts testosterone to DHT can inhibit the growth of the flank organ. Certain unsaturated aliphatic fatty acids, such as gamma-linolenic acid and myristoleic acid, as well as other natural compounds, including alizarin and curcumin, are 5alpha-reductase inhibitors and inhibited flank organ growth. Green tea catechins, including (-)-epicatechin-3-gallate, and (-)-epigallo-catechin-3 gallate (EGCG) are also 5alpha-reductase inhibitors and inhibited flank organ growth. However, (-)-epicatechin and (-)-epigallocatechin, which are not 5alpha reductase inhibitors, also inhibited flank organ growth. EGCG also inhibited DHT dependent growth of flank organs. These catechins, therefore, may act by a mechanism other than inhibition of 5alpha-reductase. The effect of EGCG and other compounds was localized at the site of application; they did not affect the growth of the contralateral flank organ in the same animal. Since these compounds do not appear to exhibit systemic effects, they may be potentially useful for treatment of androgen-dependent skin disorders. PMID- 11380154 TI - Serum factors regulate the expression of the proliferation-related genes alpha5 integrin and keratin 1, but not keratin 10, in HaCaT keratinocytes. AB - In the highly coordinated programme of gene expression during keratinocyte proliferation and differentiation, alpha5 integrin and keratins 1 and 10 (K1/K10) may play important regulatory roles. We were interested in seeing whether, in continuously growing, immortalized HaCaT keratinocytes, similar to normal keratinocytes, the expression of alpha5 integrin and K1/K10 was related to cell proliferation and differentiation. After release from cell quiescence the expression of alpha5 integrin, both at the mRNA and protein levels, was upregulated in the cells. At the same time, K1/K10 mRNA and protein expression decreased dramatically, while the mRNA for D1 cyclin became detectable, and the cells became highly proliferative. These findings indicate that alpha5 integrin and K1/K10 are involved in the regulation of HaCaT proliferation and differentiation, as in normal keratinocytes. However, HaCaT cells are different from normal keratinocytes in their ability to lose K1/K10 expression. There is no evidence that the expression of K1/K10 can be reversed in normal keratinocytes. This ability of dedifferentiation might be a unique feature of HaCaT cells and may be a key component of their immortalized nature. We also found that serum factors regulate mRNA expression of alpha5 integrin and K1, but not of K10, in HaCaT cells. This information could be relevant to the understanding of normal epidermal differentiation. PMID- 11380155 TI - The role of nitric oxide in allergic contact dermatitis. PMID- 11380156 TI - Patch test results with tixocortol pivalate and budesonide in Germany and Austria. PMID- 11380157 TI - Social deprivation and psychiatric service use for different diagnostic groups. AB - Recent research has shown that the relationship between social deprivation and admission rates varies according to diagnosis. We have replicated a study of this kind. Furthermore, in addition to admission rates, we also look at variations in length of stay and the proportion of readmitted patients. Psychiatric admission data for the 79 Amsterdam neighbourhoods was obtained from the City Psychiatric Register. This data covered all the admissions between 1992 and 1995 of people from Amsterdam aged 19 and older, with the exception of short-term crisis admissions to the Crisis Centre. These admissions were divided into six diagnostic groups. The admission rates, the average length of stay and the proportion of patients readmitted were compared to the level of socio-economic deprivation in the area concerned, a factor which was determined using factor analysis. Admission rates for schizophrenia, other psychoses and neurosis disorders showed a significant relation with the level of socioeconomic deprivation. Admission rates for affective disorders, organic psychoses and personality disorders showed no significant relation with deprivation. The findings on average length of stay and proportion of readmitted patients showed no clear relation with deprivation according to diagnosis category. As was found in previous studies, the relationship with socio-economic deprivation varies according to diagnosis. The average length of stay and the risk of readmission, given the diagnosis, do not vary according to level of deprivation. PMID- 11380158 TI - What do women want? Women's experiences of infertility treatment. AB - Finnish women's experiences of infertility treatment were investigated by examining their satisfaction and dissatisfaction, and their most positive and negative experiences with the treatment. Three hundred and forty four (16%) out of the 2,189 women respondents to a 1994 postal survey (response rate 74%) had experienced difficulties in having a baby. Two-thirds had sought medical help, generally from private gynaecologists. Less than half of the women were satisfied with the infertility treatment, expressing less satisfaction than is generally found among health care clients. Dissatisfied women were more often 35-39 years of age, in treatment during the study period, in treatment in public clinics and not successful in having a baby. However, about one-third of the women were unsure about or did not give their opinion in regard to satisfaction. The subsequent birth of a baby was the most common reason for satisfaction. The most positive treatment experience was respectful, empathic and personal care from the doctor. Unsatisfactory encounters with health care personnel were the main reasons for dissatisfaction and were most often cited as the most negative treatment experience. This dissatisfaction could reflect relatively young and healthy women's assertive attitudes toward infertility care services in the context of the intimacy and vulnerability of childlessness. PMID- 11380159 TI - The social context of health selection: a longitudinal study of health and employment. AB - Health selection out of the labour force has received considerable attention by analysts attempting to disentangle the "true" biological dimensions of ill-health from its social meaning. Rejecting this dualistic separation, we argue that the effect of health on labour force participation is an inherently social process reflecting differential access to material and symbolic rewards that are structured by social position. Using longitudinal data from the US-based Panel Study of Income Dynamics, we examine the extent to which structural arrangements, including those designated by gender, race, education and age, differentially affect the risk of a labour market exit when health is compromised. Individuals employed at entry into the study (from 1984-1990) were followed for the duration of the study or until they left the labour force. Analyses were stratified by gender and age (25-39 and 40-61 years at baseline). We found suggestive evidence that the hazard of labour market exit in the context of perceived ill-health depended on gender, race and education, but in ways that were not constant across each of these social positions. For example, men may be more vulnerable to the labour market effects of poor health, but only in the younger group, black men were less likely to leave the labour force than white men, and education mattered, but only among younger women and older men. While these patterns may reflect differential access to disability pensions or other work-related benefits, we suggest that a more detailed analysis of trajectories of health and employment. as well as the meaning of health states would be useful in further elucidating the social dimensions of health selection. PMID- 11380160 TI - Socioeconomic disparities in health change in a longitudinal study of US adults: the role of health-risk behaviors. AB - This study investigated the hypothesis that socioeconomic differences in health status change can largely be explained by the higher prevalence of individual health-risk behaviors among those of lower socioeconomic position. Data were from the Americans' Changing Lives study, a longitudinal survey of 3,617 adults representative of the US non-institutionalized population in 1986. The authors examined associations between income and education in 1986, and physical functioning and self-rated health in 1994, adjusted for baseline health status, using a multinomial logistic regression framework that considered mortality and survey nonresponse as competing risks. Covariates included age, sex, race, cigarette smoking, alcohol consumption, physical activity, and Body Mass Index. Both income and education were strong predictors of poor health outcomes. The four health-risk behaviors under study statistically explained only a modest portion of the socioeconomic differences in health at follow-up. For example, after adjustment for baseline health status, those in the lowest income group at baseline had odds of moderate/severe functional impairment in 1994 of 2.11 (95% C.I.: 1.40, 3.20) in an unadjusted model and 1.89 (95% C.I.: 1.23, 2.89) in a model adjusted for health-risk behaviors. The results suggest that the higher prevalence of major health-risk behaviors among those in lower socioeconomic strata is not the dominant mediating mechanism that can explain socioeconomic disparities in health status among US adults. PMID- 11380161 TI - Defining the "urban" in urbanization and health: a factor analysis approach. AB - Urban environments have been linked to a range of human health issues, and as the pace of urbanization accelerates, new challenges arise to characterize these environments, and to understand their positive and negative implications for health. We seek to contribute to future studies of urbanization and health by exploring multiple definitions of urbanicity in the Philippines, using data from an ongoing, longitudinal study. We use factor analysis to identify meaningful clusters of household- and community-level variables, and to generate factor scores that summarize each household's position with respect to access to infrastructure and health services, and level of affluence. Factor scores are considered for 1983 and 1994 to assess the type and pace of change that has occurred in the Philippines, and scores are compared across urban and rural areas, and across six different settlement types, to explore household- and community-level markers of urbanicity. This analysis demonstrates the heterogeneity of environments within urban and rural areas, and emphasizes the need for a finer level of investigation in future studies of urbanization and health. PMID- 11380162 TI - Casuistry as bioethical method: an empirical perspective. AB - This paper examines the role that casuistry, a model of bioethical reasoning revived by Jonsen and Toulmin, plays in ordinary moral reasoning. I address the question: 'What is the evidence for contemporary casuistry's claim that everyday moral reasoning is casuistic in nature?' The paper begins with a description of the casuistic method, and then reviews the empirical arguments Jonsen and Toulmin offer to show that every-day moral decision-making is casuistic. Finally, I present the results of qualitative research conducted with 15 general practitioners (GPs) in South Australia, focusing on the ways in which these GP participants used stories and anecdotes in their own moral reasoning. This research found that the GPs interviewed did use a form of casuistry when talking about ethical dilemmas. However, the GPs' homespun casuistry often lacked one central element of casuistic reasoning--clear paradigm cases on which to base comparisons. I conclude that casuistic reasoning does appear to play a role in every-day moral decision-making, but that it is a more subdued role than perhaps casuists would like. PMID- 11380163 TI - Intrahousehold resource allocation and child growth in Mozambique: an ethnographic case-control study. AB - This study examines the effect of intrahousehold cash income control and decision making patterns on child growth in the rural town of Sussundenga in Manica Province, Mozambique. A case-control study design was used to examine the influence of men's and women's disaggregated cash incomes on child growth. The research tested whether greater maternal share of household cash income was associated with (1) increased maternal decision-making and bargaining power in the household, and (2) better child growth. Fifty case households, with children 1-4 years old exhibiting poor growth, were matched with 50 control households of similar socioeconomic status in which all children under five demonstrated healthy growth. Data were gathered on gender-specific income generation and expenditure, specific intrahousehold allocation processes, diet, and sociodemographic variables using a formal survey. Key informant interviews, focus groups, and observation over one year provided ethnographic context for the case control findings. Case-control differences were analyzed using McNemar's test, paired t-test, and conditional logistic regression. In spite of matching households for socioeconomic status, control household incomes were still slightly greater than cases. Male spouse income was also higher among controls while maternal income, and maternal proportion of household income, were not significantly different. Household meat, fish and poultry consumption, and maternal education were significantly greater among control households than cases. Greater maternal share of household income was not associated with greater maternal decision-making around cash. However, mothers must spend what little cash they earn on daily food supplies and usually request additional cash from spouses to cover these costs. There is evidence that if mothers earn enough to cover these socially prescribed costs, they can spend cash for other needs. Above this threshold, women's earnings may confer more bargaining power. The research also revealed a nuclearization of households, attenuation of community bonds of mutual aid, and increasing importance of cash for survival. PMID- 11380164 TI - Does the state you live in make a difference? Multilevel analysis of self-rated health in the US. AB - This paper investigates the different sources of variation between US states in self-rated health using multilevel statistical procedures. The different sources that are considered are based on individual- and state-level factors. Data for the analysis comes from the 1993-94 Behavioral Risk Factor Surveillance System and the 1986-90 General Social Surveys. Results show that individual-level factors (such as low income, being black, smoking) are strongly associated with self-rated poor health. Significant variation, however, remain between states after allowing for individual characteristics. Crucially, between-state variation in self-rated health is different for different income groups. State-level contextual effects are found for per-capita median-income and 'social capital'. While not strong, there seems to be a differential impact of state income inequality on high-income groups, such that the affluent report better health from living in high inequality states. The paper substantiates the need to connect individual health to their macro socioeconomic context. Importantly, it is argued that without adopting an explicitly multilevel approach, the debate on linkages between individual health and income-inequality/social capital cannot be adequately addressed. PMID- 11380165 TI - Condom use, power and HIV/AIDS risk: sex-workers bargain for survival in Hillbrow/Joubert Park/Berea, Johannesburg. AB - Through interviews with 50 female sex-workers in the Hillbrow/Berea/Joubert Park area of Johannesburg, this paper explores sexual negotiations between men and women in the sex industry. This paper focuses on factors that affect sexual decision-making including safer sex practices. In moving beyond approaches that emphasize women's 'powerlessness' in sexual negotiation, this article focuses on ways in which sex-workers capitalize on clients' reluctance to use condoms in sexual exchanges. We emphasize sex-worker's agency and use a broader, Foucauldian understanding of power, which couples power with resistance. Further, this paper examines other elements of the sex industry that contribute to unsafe sex such as competition between women for clients and violence in the industry. Finally, this paper suggests that HIV-prevention programs take cognizance that power negotiations between men and women cannot be simplistically understood as men having power and women being powerless. Rather, this article contributes to a growing body of literature in medical anthropology, which elucidates the complexities of sexual negotiations between men and women. This focus on agency is important in trying to lessen the stigma and discrimination that sex-workers face at the hands of clients, pimps/managers, police and health care workers. PMID- 11380166 TI - Increased toxicity of textile effluents by a chlorination process using sodium hypochlorite. AB - Chlorinated textile effluents were tested for their toxicity using different bioassays. These assays were the Microtox assay, daphnia (Daphnia similis) 48-hr survival test, medaka embryo 14-day and juvenile 96-hr survival tests, and tilapia (Oreochromis mossambicus) juvenile 96-hr survival test. By comparing the results of toxicity tests on water samples collected at the instream prior to the chlorination process and at the outlet of the wastewater treatment facility, we found that wastewater toxicity was obviously increased by chlorination using NaOCl as the oxidant, as evidenced by the different bioassays used. Because no significant difference was observed in water chemistry, such as pH, DO, and conductivity, the induced-toxicity may be partially attributable to residue chlorine or other chlorinated compounds generated by chlorination. Future studies are warranted to identify the cause of the increase in the textile wastewater toxicity. PMID- 11380167 TI - Study on endocrine disrupting chemicals in wastewater treatment plants. AB - From July 1998 to March 1999, a study was made of a total of 27 treatment plants for the principal purpose of understanding the actual condition of endocrine disrupting chemicals (EDCs) in sewage, and the behavior of EDCs in wastewater treatment plants. The results showed actual levels of influent and effluent concentrations of EDCs in sewage. Substances detected above the minimum limit of determination were 15 for wastewater influent and 6 for effluent. Similarly, nonyl phenol ethoxylate and 17 beta-estradiol, which are highlighted as pertinent substances, were detected. It was confirmed that the reduction ratio of EDCs in treatment plants was 90% or more for almost all substances. The behavior of EDCs in general in treatment plants was also studied. As a result, the EDCs reduction effect was recognized in both the primary setting tank and biological reaction tank, though the trend varies among substances. PMID- 11380168 TI - Estrogenic influences of estradiol-17 beta, p-nonylphenol and bis-phenol-A on Japanese medaka (Oryzias latipes) at detected environmental concentrations. AB - Mature male medaka were continuously exposed to 0.005, 0.0-5 or 1.0 ppb of estradiol-17 beta (E2 or 0.1, 10 or 100 ppb of p-nonylphenol (NP) or bis-phenol-A (BPA). Female-specific proteins (Fsp) were induced in medaka exposed to 0.005 ppb of E2, 0.1 ppb of NP, or 10 ppb of BPA. Concentrations of 0.005 pbb of E2 and 0.1 ppb of NP corresponded to concentrations of these chemicals detected in river water in Japan. The abilities of the 3 chemicals to induce Fsp were E2 > NP > BPA. Embryonic medaka were exposed to E2, NP and BPA under conditions of static renewal for 200-230 days until pre-maturity. Survival ratios of medaka exposed to E2 and NP declined in concentrations more than 25 ppb and 50 ppb, respectively. The groups of medaka exposed to E2 had individuals with testis-ova or abnormal gonad. There was no male in exposure to 1.0 ppb E2. When exposed to 100 ppb of NP or BPA, abnormal gonad was also detected. Abnormal anal fin (female-like) was observed in male exposed to 100 ppb of NP. The LC50 values for each of the 3 chemicals were much higher than the concentrations detected in water in the environment--the 3 chemicals were considered to have no lethal effect on medaka in aquatic environments. However, exposures to E2 or NP at environmental concentrations induced Fsp. BPA also had the ability to affect medaka as an environmental estrogen, although its extrogenic activity was weaker than that of E2 or NP. PMID- 11380169 TI - Detection of estrogenic activity in Flemish surface waters using an in vitro recombinant assay with yeast cells. AB - Numerous environmental chemicals possess estrogen-like properties. At elevated doses, natural estrogens and environmental estrogen-like chemicals are known to produce adverse effects on humans and wildlife. Sources of potential exposure to endocrine disrupting compounds have to be identified for risk and hazard assessment. Extracts prepared from 16 selected water samples taken in Flemish rivers, effluents of municipal wastewater treatment plants and reservoirs for drinking water production were analysed for estrogenic activity with a cellular bioassay. Yeast cells, which are stably transfected with the DNA sequence of hER and which contain expression plasmids with the reporter gene lac-Z, encoding the enzyme beta-galactosidase, were used to measure receptor binding. Flemish rivers showed the highest estrogenic potency, compared to effluents of waste water treatment plants and reservoirs which showed low induction factors (beta galactosidase production) relative to solvent control conditions. By comparison with a standard curve for 17 beta-estradiol (E2), estrogenic potency in water samples was calculated as E2-equivalents and ranged from below detection limit (approximately 2.75 ng E2/l) up to 81.4 ng/l E2-equivalents. About 7 water samples had more than 10 ng/l E2-equivalents. These elevated levels of E2 equivalents are likely to exert significant adverse effects on reproduction success of wildlife, which should be verified with in vivo studies. PMID- 11380170 TI - Comparison between estrogenicities estimated from DNA recombinant yeast assay and from chemical analyses of endocrine disruptors during sewage treatment. AB - This study discusses the estrogenicity and the extent of estrogenic effects, of sewage and treated sewage in public sewage treatment plants in Japan. The estrogenicity in this study was measured with a DNA recombinant yeast strain. Using this method, 43 chemicals that are suspected to have estrogen-like effects were measured and their estrogenicities were evaluated in terms of 17 beta estradiol equivalents by comparison with the estrogenicity of 17 beta-estradiol. 17 beta-estradiol equivalents of influent and effluent sampled from 20 sewage treatment plants (STPs) were measured with this method. Because the concentrations of endocrine disruptors (EDs) in the STPs were monitored by the Ministry of Construction (MOC), the estrogenic effects estimated from the chemical data were obtained as a theoretical estrogenicity in terms of 17 beta estradiol equivalent. The results suggest that STPs effectively reduce the estrogenicity and the theoretical estrogenicity during treatment, and that there were some differences between the estrogenicity assayed by the yeast and the theoretical estrogenicity in many STPs, particularly in influent sewage. Therefore, it is implied that unknown estrogen-like substances or antagonists might exist in influent sewage and treated sewage in STPs. PMID- 11380171 TI - Dose-response assessment by a fuzzy linear-regression method. AB - Regression analysis has been used to characterize the relationship between an exposure dose and the incidence of an adverse health effect such as cancer. However, the regression rarely describes the true relationship due to uncertainties in dose-response data and relationships. Therefore, a method is developed to perform dose-response assessments by a fuzzy linear regression which explicitly exhibit these uncertainties. This method is applied to define the relationship between a particular nitrate dose to humans and its corresponding cancer risk. PMID- 11380172 TI - Statistical comparisons of the no-observed-effect concentration and the effective concentration at 10% inhibition (EC10) in algal toxicity tests. AB - Batch-type algal toxicity tests were conducted to evaluate the low toxic effects of metal toxicants. Both the no-observed-effect concentration (NOEC) and the effective concentration at 10% inhibition (EC10) were calculated and compared. The results of this study indicate that, for algal toxicity tests, NOEC offers better protection to the test organisms than EC10. In addition, the consistency of NOEC is found to depend on the response endpoints measured. A cut-off-value approach is proposed to determine whether NOEC or EC10 should be chosen for estimating low toxic effects. For a specific toxicity test, the average cut-off value directly indicates the magnitude of the square root of the within-group variance. A precise test associated with small within-group-variances also produces relatively small average cut-off value (say, less than 10% of the growth reduction). This average value provides useful information on the minimum protection that can be offered by the NOEC. PMID- 11380173 TI - Monitoring and classification of toxicity using recombinant bioluminescent bacteria. AB - Recombinant bioluminescent biosensing cells were used to detect and classify toxicity caused by various chemicals in water environments. Classification of the toxicity was realized based upon the chemicals' actions of toxicity by using DNA , oxidative- and membrane-damage sensitive strains. Tested samples contained a single chemical or mixture of chemicals in media, wastewater, or river water. Finally, it is suggested that this method for classification of toxic chemicals in wastewater or other aqueous systems may be adopted for primary screening steps of the samples and can give useful information about the samples' characteristics. PMID- 11380175 TI - Ecological co-inhabitance index (ECI) as a management tool for ecosystem preservation in rivers. AB - The number of novel man-made hazardous substances produced by industries is increasing year after year, resulting in conventional discharge standards ineffective to preserve the natural ecological environment. A novel index, ecological co-inhabitance index (ECI), was proposed in order to evaluate the river ecosystem sensitively. The river benthic community with a high value of ECI is formed in the healthy ecosystem where benthic animals can share resources in the river environment efficiently, keep the ecosystem functioning, and give the least adverse effect to lower reaches of the river. In order to clarify the characteristic of ECI, the relationship between ECI and water quality in the river was investigated using the data on benthic animals obtained from 17 various rivers. Results of this investigation indicated that ECI could synthetically evaluate the river environment without a bias toward a specific water quality. Moreover, ECI had the significant correlation coefficients with diversity index, biotic index and pollution index at significance level 0.05, respectively. Therefore, ECI is a promising index for managing the river ecosystem. PMID- 11380174 TI - Biomarker studies on gold-lined sea bream (Rhabdosargus sarba) exposed to benzo[a]pyrene. AB - Gold-lined sea bream (Rhabdosargus sarba) were intraperitoneally injected with 35 mg/kg benzo[a]pyrene (in DMSO), with fish administered DMSO as the solvent control. Fish were sacrificed 3 days post injection and their livers dissected for the analysis of adenosine triphosphates (ATPase), glutathione-S-transferase (GST), DNA adducts and ethyoxyresorufin-O-deethylase (EROD). Exposure to B[a]P resulted in reduced ATPase activity, elevated GST activity and DNA adduct level, but no apparent change in EROD activity. PMID- 11380176 TI - Comparison of a novel electro-Fenton method with Fenton's reagent in treating a highly contaminated wastewater. AB - This study applied a novel electrochemical process called the Fered-Fenton method to treat a highly concentrated wastewater. By combining electrochemical reduction and chemical oxidation, the process can remove organic compounds and heavy metals in a batch reactor. A PVC-stabilizer processing wastewater was treated in this investigation owing to its high heavy metal concentration (Pb = 7,500 mg/l) and high organic concentration (COD = 11,000 mg/l). The major organic component was acetate. Direct anodic oxidation showed no effect on COD removal. Fenton's method only removed 36% of COD using 4,000 mg-Fe2+/l and 28,000 mg-H2O2/l dosage. In the Fered-Fenton process, about 89% of COD was removed with 2,000 mg-Fe3+/l and 28,000 mg-H2O2/l. Furthermore, the COD removal attained an efficiency of about 98% for 56,000 mg-H2O2/l used. Results presented herein demonstrate that the Fered-Fenton method is superior to direct anodic oxidation and Fenton's method in this case. Furthermore, the changes of the intermediate compounds including acetate, oxadate, and formate during the reaction were analyzed, which provided us with the information to propose degradation reactions of the wastewater in this system. PMID- 11380177 TI - Effects of spilled oil on the tidal flat ecosystem--evaluation of wave and tidal actions using a tidal flat simulator. AB - The purpose of this study is to clarify the effects of wave and tidal actions on the penetration of spilled oil stranded on tidal flats and to evaluate the influence of the penetrated oil on seawater infiltration using tidal flat simulator. A simulator used was composed of tidal flat, wave maker, tide controlling device, temperature controlling system and computer controlling system. The infiltrations of seawater and fuel oil C into tidal flats were visualized using transparent glass beads as tidal flat sediments. Penetration behaviour of the spilled oil into the sediments was significantly different from that of seawater. Seawater infiltrated into the sediments both by wave action and tidal fluctuation, while fuel oil C penetrated by tidal movement only. The infiltration of seawater was reduced by penetrated oil. This result indicates that the penetrated oil diminishes infiltration of seawater into the sediments and thus results in the reduction in the supply of oxygen, nutrients, and organic matter to the benthic organisms in tidal flat. PMID- 11380178 TI - GREAT-ER: a new tool for management and risk assessment of chemicals in river basins. Contribution to GREAT-ER #10. AB - The GREAT-ER (Geo-referenced Regional Exposure Assessment Tool for European Rivers) project team has developed and validated an accurate aquatic chemical exposure prediction tool for use within environmental risk assessment schemes. The software system GREAT-ER 1.0 calculates the distribution of predicted environmental concentrations (PECs) of consumer chemicals in surface waters, for individual river stretches as well as for entire catchments. The system uses an ARC/INFO-ArcView (ESRI) based Geographical Information System (GIS) for data storage and visualization, combined with simple mathematical models for prediction of chemical fate. At present, the system contains information for four catchments in Yorkshire, one catchment in Italy, and two in Germany, while other river basins are being added. Great-ER 1.0 has been validated by comparing simulations with the results of an extensive monitoring campaign for two 'down the-drain' chemicals, i.e. the detergent ingredients boron and Linear Alkylbenzene Sulphonate (LAS). GREAT-ER 1.0 is currently being expanded with models for the terrestrial (diffuse input), air and estaurine compartments. PMID- 11380179 TI - Predicting soil-water partition coefficients for Hg(II) from soil properties. AB - The metal adsorption characteristics for fifteen Taiwan soils by Hg(II), were evaluated using pH as the major variable. The soil samples were thoroughly characterized for their physical chemical properties and composition, particularly organic matter and metal oxides. The adsorption of Hg(II) increased with increasing pH between pH 2.5 and 5.5, whereas the adsorption significantly decreased above around pH 5.5. Below pH 5.5, greater adsorption was found for soils with a higher organic matter content at constant pH and metal concentration. To better understand the mechanism of adsorption, the experimental results for Hg (II) were tested in a partition coefficient model to relate the adsorption of the Hg(II) by the different soils with soil components: organic matter, iron oxide, aluminium oxide and manganese oxide. This model was not successful when applied to measurements at the differing natural soil pHs because of the importance of pH. At pH greater than 5.5 the model fails because of the complexation of Hg by the dissolved organic matter. However, partition coefficients obtained from experimental data were highly correlated with those calculated for a partition coefficient between mercury and organic matter alone at lower pH. Normalization of the partition coefficients, Kd, for the organic matter content of the soils, Kom, greatly improved the correlation between the partition coefficient and pH under pH 5.5 (R2 increased from 0.484 to 0.716). This suggests that the surficial adsorption sites are principally due to organic matter for pH less than 5.5. For the 24-hour equilibration period employed, diffusion of Hg through this superficial organic matter coating to underlying sorptive materials, including metal oxides, is not important in the partitioning of Hg. At pH above 5, a decrease of mercury adsorption with increasing solution pH was also found. This result may be explained in part by the complexation of mercury by soil dissolved organic matter whose concentration increased with increasing pH. PMID- 11380180 TI - Ozone treatment for biorefractory COD removal. AB - A two case study of industrial wastewater treatment is reported in order to prove the suitability of ozonation for biorefractory COD removal. The ozone application as an end treatment is shown to provide an excellent oxidation yield of organic matter in a landfill leachate. The combination of ozonation and fixed bed biofilm process appears quite effective for the post-treatment of two biological effluents from paper industry. PMID- 11380181 TI - Investigation of radical reactions for efficiency improvement in ozone-hydrogen peroxide treatment. AB - It was verified by laboratory-scale experiments that the two-port ozone injection method was more advantageous for reaction efficiency in the ozone-hydrogen peroxide treatment than the conventional one-port ozone injection method under the condition of same ozone dose. The reason for this effectiveness was also investigated in detail by computer simulations in the wide range of the rate constant of OH radical reacting with TOC (kTOC). The laboratory-scale experiments revealed that the amount of TOC decomposed (delta TOC) increased by up to 41% by the two-port injection method, indicating a 27% improvement in delta O3/delta TOC. A detailed investigation of these experimental results by computer simulations showed that by injecting ozone in two ports the amount of ozone consumed for the OH radical formation was increased more remarkably than that of ozone consumed for the radical chain reactions, and that the amount of OH radical reacting with TOC was increased more strikingly than that with ozone. Furthermore, it was found that the smaller the kTOC value with 10(5)-10(9) M-1s-1 was, the more advantageous for the reaction efficiency the two-port injection method was. PMID- 11380182 TI - Heterogeneous catalytic ozonation of 2-chlorophenol aqueous solution with alumina as a catalyst. AB - Heterogenous catalytic ozonation of 2-chlorophenol (2-CP) in the presence of gamma-alumina as a solid catalyst has been investigated in this research. It showed that the rate for degradation of TOC could increase from 21% to 43%. The pseudo-first reaction constants of 2-CP could increase from 0.8688 min-1 to 0.1270, increasing by approximately 40%. At the same time, the consumption of ozone was only half that of ozone alone. This research also explored the effects of the catalyst dosage, pH values and removal kinetics of 2-CP. In addition, three consecutive running with the same catalyst revealed insignificant reduction of the activity. Furthermore, the elimination of toxicity was evaluated by Microtox analysis. The detoxification was more stable and with good results. PMID- 11380184 TI - Catalytic wet air oxidation of p-nitrophenol (PNP) aqueous solution using multi component heterogeneous catalysts. AB - This study investigated the decomposition of high strength p-nitrophenol (PNP) of 2,000 mg/l (3,400 mg of COD/1,250 mg of TOC) by catalytic wet air oxidation. Multi-component heterogeneous catalysts were used as catalysts for this purpose. The study results using a batch reactor showed that catalyst "D" (Mn-Ce-Zr 22.4 g plus CuSO4 1.0 g; Mn-Ce-Zr-Cu [CuSO4]) was more effective (56-74%) than catalyst "A" (Mn-Ce-Zr 22.4 g) under the given conditions (O2 partial pressure of 1.0 MPa; temperature of 170-190 degrees C; 30 min of reaction time). The best result was obtained when 2 g of Mn-Ce-Zr-Cu [CuSO4] was used per 1L of PNP aqueous solution. COD and TOC removal efficiencies were 18% and 23% without catalysts during 20 min of reaction at 190 degrees C. They were improved to 79% and 71% with 2 g/L of Mn Ce-Zr-Cu [CuSO4] under the same conditions. The ratio of BOD5/COD was measured to evaluate biodegradability. It was 0.05 without catalyst and increased to 0.33 with 2 g/L of Mn-Ce-Zr-Cu [CuSO4] for 20 min of reaction. PMID- 11380183 TI - Partial oxidation of reactive dyestuffs and synthetic textile dye-bath by the O3 and O3/H2O2 processes. AB - Ozonation and O3/H2O2 oxidation of reactive dyestuffs and simulated textile dye bath were investigated. Effects of reaction pH, initial dye concentration, H2O2 concentration and assisting chemicals on treatment efficiency were examined. We found considerable improvement in COD and colour removal rates at pH = 11, that was almost the actual pH of the prepared textile wastewater, whereas in particular increasing the initial dyestuff concentration had an adverse effect upon oxidation rates. Removal of colour, COD and TOC were found to be fairly sensitive to the introduction of soda that is frequently applied as an auxiliary chemical during the reactive dyeing process. The addition of H2O2 had negligible effect on COD removal efficiency and decolorization rate compared to ozonation alone at different pH values. Accordingly it can be inferred that the theoretically expected effect of OH radical scavengers (e.g. carbonate, chloride) present in the synthetic dye-bath as well as introduced radical formation promoters (e.g. H2O2, OH-) were probably hidden due to the complexity of the synthetic wastewater matrix. Biodegradability of the ozonated dye-bath was accelerated by a factor of three corresponding to a 233% relative enhancement. The inhibition of the oxygen uptake rate decreased from 91% to 75% within only 5 min treatment time. PMID- 11380185 TI - Ion exchange selectivities of calcium alginate gels for heavy metals. AB - An equilibrium model has been proposed and verified, based on the conditions in the gel phase and Donnan equilibrium theory, for the analysis of the experimental data on the recovery of lead, copper, cadmium, cobalt, nickel and zinc from synthetic, nonmetallic aqueous solutions on calcium alginate gels. This equilibrium model considers that the system behaves as an ion-exchange process between the calcium in the gels and the divalent metals in solution, and that the metallic portion enclosed in gel fluid is supposed an important quantitative contribution to the total amount of metal uptake by gels. According to the equilibrium constants calculated, it is deduced that the selectivity order is: Pb > Cu > Cd > Ni > Zn > Co. PMID- 11380186 TI - The potential application of activated carbon from sewage sludge to organic dyes removal. AB - The objective of this research work was to study the potential application of activated carbon from sewage sludge to organic dye removal. Methylene blue and crystal violet were the two dyes investigated in the present study. Three activated carbons were produced from the exclusive sewage sludge (referred to as DS), the sludge with the additive of coconut husk (DC) and sludge with the additive of peanut shell (DP) respectively. They were characterized by their surface area and porosity and their surface chemistry structure. Adsorption studies were performed by the batch technique to obtain kinetic and equilibrium data. The results show that the three sludge-derived activated carbons had a developed porosity and marked content of surface functional groups. They exhibited a rapid three-stage adsorption process for both methylene blue and crystal violet. Their adsorption capacities for the two dyes were high, the carbon DP performed best in the adsorption whereas the carbon DC performed worst. It is therefore concluded that the activated carbons made from sewage sludge and its mixtures are promising for dye removal from aqueous streams. PMID- 11380187 TI - RDX degradation using an integrated Fe(0)-microbial treatment approach. AB - RDX is a persistent and highly mobile groundwater contaminant that represents a major remediation challenge at numerous munitions manufacturing and load assemblage-package facilities. This work presents proof of concept that permeable reactive iron barriers might be a viable approach to intercept and degrade RDX plumes. Specifically, RDX was rapidly reduced in aquifier microcosms amended with Fe(0) powder, and in flow-through columns packed with steel wool. The rate and extent of RDX degradation in microcosms was enhanced by anaerobic bacteria that feed on cathodic hydrogen (i.e., H2 produced during anaerobic Fe(0) corrosion by water). Apparently, the hydrogenotrophic consortium that exploits Fe(0) corrosion as a metabolic niche participated in the further degradation of heterocyclic intermediates produced by the reaction of RDX with Fe(0). Reductive treatment of RDX with Fe(0) also reduced its toxicity to microorganisms and enhanced its subsequent biodegradability under either anaerobic or anaerobic conditions. Therefore, a combined or sequential Fe(0)-biological treatment approach might improve treatment efficiency. PMID- 11380188 TI - Peroxidase-catalyzed removal of phenols from a petroleum refinery wastewater. AB - The phenol content of a petroleum refinery wastewater was reduced below the discharge limit following treatment with horseradish peroxidase and H2O2. Approximately 58% of COD, 78% of BOD5, and 95% of toxicity were removed along with the phenols. As a result of treatment, phenols were transformed into less biodegradable compounds which could be removed by subsequent coagulation and precipitation. Optimization of the peroxide concentration led to 20% enzyme savings. The use of PEG and chitosan as protective additives resulted in 4 and 25 fold reductions in enzyme requirements, respectively. Phenol removal did not appear to be adversely affected by the presence of other hydrocarbons that are frequently present in refinery wastewaters. PMID- 11380189 TI - Kinetics of azoreductase and assessment of toxicity of metabolic products from azo dyes by Pseudomonas luteola. AB - This is a continuous study on a decolorization strain, Pseudomonas luteola, which involves treating seven azo dyes with different structures. This study focuses mainly on determining both the mechanism of decolorization by P. luteola and the activity of azoreductase from P. luteola as well as identifying and assessing the toxicity of metabolic products of azo dyes. The growth of P. luteola reached the stationary phase after shaking incubation for 24 hours. Then, while being kept static, the color of seven tested azo dyes (100 mg/l) could be removed. The proportion of color removal was between 59-99%, which figure is related to the structure of the dye. Monoazo dyes (RP2B, V2RP and Red 22) showed the fastest rate of decolorization, i.e. from 0.23-0.44 mg dye-mg cell-1 hr-1. P. luteola could remove the color of V2RP and a leather dye at a concentration of 200 mg/l, and as to the rest of the azo dyes, it could remove at a concentration of up to 100 mg/l. Decolorization of RP2B and Red 22 required activation energy of 7.00 J/mol and 6.63 J/mole, respectively, indicating that it was easier for azoreductase to decolorize structurally simple dyes. The kinetics of azoreductase towards seven azo dyes suggested a competitive inhibition model be applied. Microtox was used to analyze the toxicity of the metabolic products of azo dyes. EC50 showed differences in toxicity before and after the azo dyes had been metabolized. Analysis revealed significant differences between the results obtained by EC50 with Blue 15 and those obtained with the leather dye, indicating that the toxicities of the metabolic products were increased. The differences obtained by EC50 with Red 22, RP2P and V2RP were small, and Black 22 showed no such difference. Sulfanic acid and orthanilic acid may be the intermediate products of Violet 9 and RP2B, respectively. However, according to FT-IR analysis, aromatic amines were present in the metabolic product. PMID- 11380190 TI - Biotreatability studies of pharmaceutical wastewater using an anaerobic suspended film contact reactor. AB - The pharmaceutical industrial effluents, which include several organic solvents and other toxic chemicals, are generally treated by aerobic process, which is cost intensive in nature. The alternative anaerobic route to degrade the toxic effluents is attractive due to the lower cost of treatment and the generation of gas, which can supplement the energy requirements. There are few reports on the anaerobic treatment of the pharmaceutical effluents. In the present investigation, the effluents from a bulk drug industry, which utilizes several organic chemicals, have been taken to assess their applicability for anaerobic treatment. The organic loading rates were varied from 0.25 kg/m3/day to 2.5 kg/m3/day and the COD reduction was found to be in the range of 60 to 80%. Long term operation of an anaerobic suspended film contact reactor carried out with 1.25 kg/m3/day was found to be optimum. The biogas generated during the degradation process was monitored and the methane content was found to be 60-70%. PMID- 11380191 TI - Aerobic biodegradation of gasoline oxygenates MTBE and TBA. AB - MTBE degradation was investigated using a continuously stirred tank reactor (CSTR) with biomass retention (porous pot reactor) operated under aerobic conditions. MTBE was fed to the reactor at an influent concentration of 150 mg/l (1.70 mmol/l). A second identical reactor was operated as a control under the same conditions with the addition of 2.66 g/l of sodium azide, to kill any biological activity. Results from these experiments suggest that biomass retention is critical to the degradation of MTBE. The rate of MTBE removal was shown to be related to the VSS concentration. MTBE removal exceeded 99.99% when the VSS concentration in the reactor was over 600 mg/l. Results obtained from batch experiments conducted on mixed liquor samples from the porous pot reactor indicate that the individual rates of biodegradation of MTBE and TBA were higher for initial concentrations of 15 mg/l than for concentrations of 5 mg/l. The presence of TBA at lower concentrations did not effect the rate of MTBE degradation, however higher concentrations of TBA did reduce the rate of biodegradation of MTBE. Denaturing Gradient Gel Electrophoresis (DGGE) analysis reveals that the culture consisted of a community of bacterial organisms of about 6 species. PMID- 11380192 TI - Fungal mediated decolorization of media containing procion dyes. AB - An isolated fungus, Aspergillus foetidus was found to effectively decolorize media containing azo reactive dyes, namely Procion dyes, under aerobic conditions. The extent of colour removal was 90% within 48 h of growth of the fungus. The entire colour was found to be strongly bioadsorbed to the rapidly settling spherical fungal biomass pellets. Our investigations reveal that the process of decolorization is concomitant with the exponential growth phase of the fungus and has an obligate requirement for a biodegradable substrate such as glucose. Kinetic analyses of fungal decolorization indicate that the rates of colour uptake decrease to a significant extent with increasing initial concentrations of dye. The fungus was able to grow and decolorize media in the presence of 5 ppm of chromium and 1% sodium chloride. An alternate and cheaper carbon source such as starch supported the growth and decolorization process. These results suggest that the dye uptake process mediated by Aspergillus foetidus has a potential for large-scale treatment of textile mill discharges. PMID- 11380193 TI - Effect of compost in phytoremediation of diesel-contaminated soils. AB - The effect of compost on phytoremediation of diesel-contaminated soils was investigated using 130 small (200 g) containers in two screening tests. The experiments were conducted in a controlled environment using ryegrass from seed. Containers were destructively sampled at various times and analyzed for plant mass and total petroleum hydrocarbons. The results indicate that the presence of diesel reduces grass growth, and that compost helps reduced the impact of diesel on grass growth. The addition of compost helps increase diesel loss from the soils both with and without grass, though the addition of grass leads to lower diesel levels compared with controls. A second set of experiments indicates that the compost helps in phytoremediation of diesel-contaminated soil independent of the dilution effect that compost addition has. The results indicate that the compost addition allowed diesel loss down to 200 mg TPH/kg even though the compost would be expected to hold the diesel more tightly in the soil/compost mixture. The simplicity of the screening tests led to difficulties in controlling moisture content and germination rates. The conclusion of the research is that the tilling of compost into soils combined with grass seeding appears to be a valuable option for treating petroleum-contaminated soils. PMID- 11380194 TI - Removal of chlorinated pollutants by a combination of ultrasound and biodegradation. AB - Chlorinated organic compounds are hazardous pollutants found in waste water, surface water, and ground water. Our study shows that a combination of ultrasonic pretreatment and biodegradation effectively removes the solvent chlorobenzene and the disinfectant 2,4-dichlorophenol, also reduces Adsorbable Organic Halogens (AOX) and Chemical Oxygen Demand (COD). In our experiments, the ultrasonic dechlorination was not influenced by the presence of other soluble organic compounds like acetate or glucose. Dechlorination of chlorobenzene by ultrasound did not lead to toxic or inhibiting reaction products. More than that, the ultrasonic pretreatment significantly reduced the toxicity of 2,4-dichlorophenol and biological activity was initiated after sonication. Residual organic pollutants after ultrasonic pretreatment were eliminated by biodegradation. PMID- 11380195 TI - Removal of hydrophobic dyestuff from dyeing wastewater by photo-sensitization process. AB - Recently, photochemical reaction became more important in view of using UV in textile dyeing wastewater treatment processes, in which neither chemical sludges nor toxic residues are left in the treated effluent. The photodegradation of hydrophobic dye (Palanil Yellow 5R, PY-5R) in the presence of acetone, which performs as a solvent and/or a photo-sensitizer, was investigated in this study. The results demonstrated that photochemical reaction in the presence of acetone could rapidly and effectively enhance color removal at a wavelength of 253.7 nm. The photodegradation follows pseudo first-order decay. The rate constants and decay quantum yields of dye degradation by UV depend on the solution pH and solvent system, (i.e., acetone to water ratio). The photosensitization of the disperse dye was found to be optimized at pH 9 and in 0.5 (v:v) acetone-water ratio. PMID- 11380196 TI - Photocatalytic degradation of azo dye in TiO2 suspended solution. AB - The photocatalysis of azo dye, Orange G, by P-25 anatase TiO2 was investigated in this research. The experiments were conducted in a batch reactor with TiO2 powder suspension. Four near-UV lamps were used as the light source. The experimental variables included solution pH level, amount of TiO2, illumination light intensity, and reaction time. A pseudo-first order reaction kinetic was proposed to simulate the photocatalytic degradation of Orange G in the batch reactor. More than 80% of 10 mg/L Orange G decomposition in 60-minute reaction time was observed in this study and fast decomposition of Orange G only occurred in the presence of both TiO2 and suitable light energy. Faster degradation of Orange G was achieved under acid conditions. The degradation rates of Orange G at pH = 3.0 were about two times faster than those at pH = 7.0. Faster degradation of azo dye was observed for greater irradiated light intensity and more TiO2 present during the reaction. The reaction rates were proportional to TiO2 concentration and light intensity with the power order of 0.726 and 0.734, respectively. PMID- 11380197 TI - Kinetics and toxicity of direct reaction between ozone and 1,2-dihydrobenzene in dilute aqueous solution. AB - The presence of toxic aromatic organic compounds in industrial wastewater affects the efficiency of conventional biological treatment. The oxidizing power of ozone represents an interesting pretreatment to reduce toxicity and increase biodegradability. At low pH, ozone is known to attack electron-rich structures by direct electrophilic reactions which open aromatic rings, and generate smaller oxidized compounds. This paper reports experimental results on kinetic and toxicity aspects of ozone direct reactions with 1,2-dihydrobenzene. This toxic compound is frequently found in cellulose bleaching effluents. Although the reaction pathway is rather complex, 4-carbon compounds, such as maleic acid, are readily formed during the first stage of ozonation. These 4-carbon compounds are further oxidized to form smaller molecules (mostly 2-carbon, such as oxalic acid). The apparent kinetics of 1,2-dihydrobenzene follows a second order law, with a rate constant around 0.36 (dm3 mmol-1s-1), at pH 2 and 20 degrees C. Results show that the BOD:COD ratio increase five-fold as ozonation progresses. Furthermore, the mutagenicity of 1,2-dihydrobenzene totally disappears as the aromatic compound is destroyed by ozonation. PMID- 11380198 TI - Practical energy and water management through pinch analysis for the pulp and paper industry. AB - In this paper we briefly describe pinch technology as a practical tool for effective energy management in the pulp and paper industry. Results indicate good steam savings. Recently pinch technology has been extended to water management. We have developed a customized methodology for the pulp and paper industry, to eliminate or reduce fresh water intake. Although the methodology is not fully developed it is a "proof of concept" that pinch principles can be applied to water related problems. The eventual combination of both thermal and water pinch will thus provide a structured and comprehensive approach for plant wide efficiency increase. PMID- 11380199 TI - Decolorization of a reactive copper-phthalocyanine dye under methanogenic conditions. AB - The objective of this research was to assess the biological decolorization of the copper-phthalocyanine dye Reactive Blue 7 (RB7) under methanogenic conditions using a mixed, methanogenic culture in a repetitive dye addition batch assay. The initial rate of decolorization was 13.2 mg/L-d and 5.7 mg/L-d for the first and second dye addition, respectively. For an initial RB7 concentration of ca. 300 mg/L, the extent of decolorization remained constant (about 62%) for each repetitive RB7 addition and resulted in a residual color build up. Declining absorbance ratio values (A664/A620) with increasing incubation time confirmed that the observed color removal was due to transformation as opposed to adsorption on the biomass. Chemical decolorization assays using sodium dithionite as the reducing agent resulted in similar absorbance spectra to that obtained after biological decolorization. In addition, in both the chemical and biological decolorization assays, partial oxidation of the reduced dye solution upon exposure to air resulted in higher residual color, indicating that the reduction and decolorization of RB7 are partially reversible. These results also suggest that RB7 reduction and decolorization both chemically and biologically most likely followed a similar reduction mechanism. PMID- 11380200 TI - Removal and recovery of heavy metals from wastewaters by supported liquid membranes. AB - The removal and recovery of Cu, Cr and Zn from plating rinse wastewater using supported liquid membranes (SLM) are investigated. SLMs with specific organic extractants as the liquid membrane carriers in series are able to remove and concentrate heavy metals with very high purity, which is very promising for recycling of heavy metals in the electroplating industry. A technical comparison between the membrane process and the conventional chemical precipitation process was made. PMID- 11380201 TI - Acrylonitrile removal from synthetic wastewater and actual industrial wastewater with high strength nitrogen using a pure bacteria culture. AB - A gram-negative rod-shaped bacteria (strain AAS6), capable of utilizing acrylonitrile as the sole source of both carbon and nitrogen, was utilized to investigate the removal of acrylonitrile in ABS resin manufacturing wastewater. Both synthetic wastewater, containing a high concentration of acrylonitrile, and actual wastewater obtained from an ABS manufacturing factory were used. The result indicated that strain AAS6 was capable of completely removing acrylonitrile from synthetic wastewater containing less than 889 mg/l acrylonitrile and from actual industrial wastewater containing less than 400 mg/l acrylonitrile. Whether in synthetic wastewater or actual industrial wastewater, strain AAS6 showed approximately the same ability for acrylonitrile removal and used acrylic acid, a metabolic by-product of acrylonitrile, as the carbon source and ammonium as the nitrogen source. The bacteria could not directly metabolize other chemicals found in the actual industrial wastewater. However, its metabolic activities were not inhibited by the presence of compounds such as butadiene, styrene or acrylonitrile-styrene polymer. Thus, this strain is expected to play an important role in aeration tanks for treating ABS resin manufacturing wastewater. PMID- 11380202 TI - Anaerobic bioventing of unsaturated zone contaminated with DDT and DNT. AB - Initial degradation of highly chlorinated compounds and nitroaromatic compounds found in munition waste streams is accelerated under anaerobic conditions followed by aerobic treatment of the degradation products. The establishment of anaerobic environment in a vadose zone can be accomplished by feeding appropriate anaerobic gas mixture, i.e., "anaerobic bioventing". The gas mixture contains an electron donor for the reduction of these compounds. Lab scale study was conducted to evaluate potential of anaerobic bioventing for the treatment of an unsaturated zone contaminated with 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) and 2,4-dinitrotoluene (DNT). Hydrogen was used as the electron donor. Using the soil columns innoculate with anaerobic microorganisms, it was observed that by feeding a gas mixture of 1% hydrogen, 1% carbon dioxide and nitrogen, methanogenic conditions were established and DDT was reductively dechlorinated. 1,1-dichloro-2,2-bis(p-chlorophenyl)ethane (DDD) accumulated as the intermediate product. The half life of DDT was calculated to be 8.5 months. DNT completely disappeared after six months of operation and no intermediates could be detected. PMID- 11380203 TI - Comparison of dye wastewater treatment by normal and anoxic + anaerobic/aerobic SBR activated sludge processes. AB - In this research, the effects of different color intensities, anoxic + anaerobic times and types of substrate on color removal effectiveness were studied using 16 litre bench scale sequencing batch reactor (SBR) anoxic + anaerobic/aerobic models. The systems were fed with a synthetic wastewater containing 500 mg/l of COD, 50 mg/l of N, 15 mg/l of P along with 10, 40 and 80 mg/l of reactive diazo Remazol Black B dye. The anoxic + anaerobic/aerobic periods were 0/11 (normal aerobic SBR), 2/9, 4/7 and 8/3 hours, respectively. The color removal occurred mainly under the anaerobic environment, while a slight attenuation was noticed under the aerobic condition. A better decolorization could be achieved with a longer anoxic + anaerobic period. The sodium acetate was a slightly better co substrate for the color removal than glucose. The decolorization efficiency dwindled as the fed dye concentration increased, but the overall color removal leaned in the opposite direction. The phosphorus removal of the anoxic + anaerobic/aerobic systems was outstanding, whereas only 50% removal was achieved for the aerobic process. A dye concentration of up to 80 mg/l did not have any significant impact on the performance of the polyphosphate accumulating organisms (PAOs). The removal of organic carbon and TKN was comparable among all experiments regardless of system configurations, color intensities and types of substrate. PMID- 11380204 TI - Development of hexachlorobenzene-dechlorinating mixed cultures using polysorbate surfactants as a carbon source. AB - The use of three nonionic polysorbate surfactants--Tween 60, 61 or 65--as the sole carbon source to sustain methanogenesis and dechlorination, as well as the effect of long-term exposure of enriched cultures to these surfactants, was investigated through the development of three sediment-derived cultures. Over a one-year period, the carbon source in these cultures was gradually switched from glucose and methanol to surfactant only, while the surfactant concentration was increased from an initial concentration of 100 mg/L to 400 mg/L. In each feeding cycle, the surfactants were partially degraded and converted to methane. Transition from glucose to Tween surfactants as the electron donor did not affect the rate, extent, and pathway of HCB transformation. These surfactants sustained the reductive dechlorination of HCB even after one year of continuous addition to the enriched cultures. This study demonstrated that reductive dechlorination of HCB sustained by the fermentation of Tween surfactants is feasible. The results support the use of anaerobically degradable Tween surfactants for the biotransformation of polychlorinated organic compounds. In principle, these surfactants could be used to simultaneously increase the bioavailability of subsurface contaminants while serving as the carbon and electron source for microbial reductive dechlorination. PMID- 11380205 TI - Changes in the organic composition of wastewater during biological treatment as studied by NMR and IR spectroscopies. AB - Despite its importance, relatively little is known about the composition and fate of wastewater organic matter (OM) in treatment plants. Monitoring the chemical changes in OM during activated sludge treatment can improve our knowledge of the processes involved in the biological elimination of OM. Direct chemical analyses of treated water OM typically account for about 20% of the OM, and structural information was obtained in this study using Nuclear Magnetic Resonance (NMR) and Fourier Transform Infrared (FTIR) spectroscopic techniques. Distinct changes in the OM during wastewater biological treatment were underlined. 13C and 1H NMR showed that aromatic carbons were minor constituents of the samples. Alkyl chains exhibited a more highly branched character in treated water, as compared to long chain aliphatic carbons present in wastewater. Carboxyl signals in the 13C NMR spectrum of wastewater could be due to peptide bonds in proteins, whereas in the treated water spectrum, this signal could be related to the presence of non proteinaceous nitrogen. Besides the non-degraded compounds, treated water OM could contain recondensation products of simple molecules. Their refractory character probably derives from their complex structures rather than from particular chemical functions, as suggested by the lack of fundamental differences in the chemical structures of wastewater and treated water OM. PMID- 11380206 TI - Biological leaching of heavy metals from anaerobically digested sewage sludge using indigenous sulfur-oxidizing bacteria and sulfur waste in a closed system. AB - The utilization of indigenous sulfur-oxidizing bacteria and sulfur waste was investigated in order to remove heavy metals from anaerobically digested sewage sludge economically. Indigenous sulfur-oxidizing bacteria existing in anaerobically digested sewage sludge were activated by adding elemental sulfur to the sludge and then the bacteria were isolated. It was found that indigenous sulfur-oxidizing bacteria could utilize sulfur waste generated by desulfurization of digestion gas as a substrate. Then, biological leaching of heavy metals from anaerobically digested sewage sludge was carried out using indigenous sulfur oxidizing bacteria and sulfur waste. By adding sulfur waste to sewage sludge, sulfuric acid was produced by the bacteria and the sludge pH decreased. Heavy metals in sewage sludge were effectively removed owing to the decrease of pH. The optimum amount of sulfur waste added to decrease the pH sufficiently was 5g/L when the sludge concentration was 2%. It was presented that the biological leaching of heavy metals from sewage sludge can be carried out in a closed system, where all required materials are obtained in a sewage treatment plant. PMID- 11380207 TI - On the contribution of background sources to the heavy metal content of municipal sewage sludge. AB - The role of human excretion, drinking water, and deposition as a source of heavy metals to municipal sewage was investigated and compared with common levels in sludge for soil application. These sources contributed more than half of the copper (Cu), zinc (Zn), and lead (Pb) content of municipal sewage sludge for soil application, while other sources dominated the fluxes of chromium (Cr) and cadmium (Cd). Drinking water was an important source for Cu and Zn. Deposition contributed about 40% to the Pb flux. Faecal excretion commonly caused less than 10% of the heavy metal load, while urinary excretion was here a negligible heavy metal source. PMID- 11380208 TI - Enhanced virus recovery from municipal sewage sludge with a combination of enzyme and cation exchange resin. AB - There is a great difficulty in virus enumeration in sewage sludge because viruses in sludge are firmly captured by sludge solids. In order to determine the precise number of viruses in sludge, an enhanced virus recovery method with a combination of an enzyme and a cation exchange resin (CER) was developed. Test viruses were seeded to a sample sludge obtained from a municipal wastewater treatment plant, and the sludge were incubated with various eluents. The quantity of eluted viruses in the liquid phase was then measured by the plaque assay technique. Using the eluent containing only water, CER, and CER with enzyme exhibited 0%, 19% and 39% of virus recovery, respectively. While the conventional USEPA method exhibited a virus recovery of 21%. Furthermore, viruses eluted by the eluent containing the CER and the lysozyme included not only surface-attached viruses but also solids-embedded viruses. PMID- 11380209 TI - Field monitoring of toxic organic pollution in the sediments of Pearl River estuary and its tributaries. AB - Field monitoring of the toxic organic compounds (PCBs, PAHs, organochlorine pesticides) in the top sediments of Pearl River Estuary and its up-streams were made. It was found that the highest concentrations of these toxic organic compounds occurred in the sediment sampled at Macau inner harbor (ZB013), which is a sink of suspended fine particles transported from the upstream waterways. Because of the affinity of the hydrophobic organic compounds (PAHs, PCBs) for the solid phase, these fine particle depositions led to accumulation of these compounds in the sediment of Macau. The atmospheric dry deposition may be another source of the toxic organic pollution in the sediment. PMID- 11380210 TI - The reactivity of natural organic matter to disinfection by-products formation and its relation to specific ultraviolet absorbance. AB - Five natural waters with a broad range of DOC concentrations were fractionated using various coal- and wood-based granular activated carbons (GAC) and alum coagulation. Adsorption and alum coagulation fractionated NOM solutions by preferentially removing components having high specific ultraviolet absorbance (SUVA). UV absorbing fractions of NOM were found to be the major contributors to DBP formation. SUVA appears to be an accurate predictor of reactivity with chlorine in terms of DBP yield; however, it was also found that low-SUVA components of NOM have higher bromine incorporation. SUVA has promise as a parameter for on-line monitoring and control of DBP formation in practical applications; however, the effects of bromide concentration may also need to be considered. Understanding how reactivity is correlated to SUVA may allow utilities to optimize the degree of treatment required to comply with DBP regulations. The reactive components that require removal, and the degree of treatment necessary to accomplish this removal, may be directly obtained from the relationship between SUVA removal and the degree of treatment (e.g., alum dose). PMID- 11380211 TI - Possible impact of treated wastewater discharge on incidence of antibiotic resistant bacteria in river water. AB - Escherichia coli and coliform group bacteria resistant to seven antibiotics were investigated in the Tama River, a typical urbanized river in Tokyo, Japan, and at a wastewater treatment plant located on the river. The percentages of antibiotic resistance in the wastewater effluent were, in most cases, higher than the percentages in the river water, which were observed increasing downstream. Since the possible increase in the percentages in the river was associated with treated wastewater discharges, it was concluded that the river, which is contaminated by treated wastewater with many kinds of pollutants, is also contaminated with antibiotic resistant coliform group bacteria and E. coli. The percentages of resistant bacteria in the wastewater treatment plant were mostly observed decreasing during the treatment process. It was also demonstrated that the percentages of resistance in raw sewage are significantly higher than those in the river water and that the wastewater treatment process investigated in this study works against most of resistant bacteria in sewage. PMID- 11380212 TI - Cutaneous stings from Bartholomea annulata. PMID- 11380213 TI - High rate of detection of unsuspected distant metastases by pet in apparent stage III non-small-cell lung cancer: implications for radical radiation therapy. AB - PURPOSE: Most radical radiotherapy (RT) candidates with non-small-cell lung cancer (NSCLC) have Stage III disease and ultimately die with distant metastases. We tested the hypothesis that positron emission tomography (PET) using 18-F fluorodeoxyglucose (FDG) would detect more unsuspected metastases in apparent Stage III disease than in Stages I-II. METHODS AND MATERIALS: Staging FDG-PET was performed for 167 NSCLC patients, with Stage I-III by conventional workup, who were candidates for curative therapy with surgery (n = 8), radical chemo/RT or RT (n = 156), or preoperative chemo/RT (n = 3). Each patient was allocated a conventional "pre-PET stage" and a "post-PET stage" that relied on PET when discordance with conventional staging occurred. RESULTS: Stage distribution pre PET was n = 39 (Stage I), n = 28 (Stage II), and n = 100 (Stage III). In 32 patients (19%), PET detected distant metastasis, most commonly abdominal with 17 cases (adrenal, n = 7; liver, n = 4; other, n = 6). Other sites included lung (n = 10) and bone (n = 6). PET-detected metastasis increased with increasing pre-PET stage from I (7.5%) through II (18%) to III (24%, p = 0.016), and, in particular, was significantly higher in Stage III (p = 0.039). Biopsy confirmation was not routine, but progression occurred at PET-detected metastatic sites or other metastatic sites in all but 3 of the 32 patients by last review. CONCLUSION: PET staging is recommended for radical RT candidates with NSCLC. The highest yield of unexpected distant metastases is observed in Stage III. PMID- 11380214 TI - Influence of interfraction interval on the efficacy and toxicity of hyperfractionated radiotherapy in combination with concurrent daily chemotherapy in stage III non-small-cell lung cancer. AB - PURPOSE: To investigate the influence of the interfraction interval (IFI) on treatment outcome and toxicity in hyperfractionated (HF) radiotherapy (RT) for Stage III non-small-cell lung cancer. METHODS AND MATERIALS: Data for 301 patients treated with 1.2 Gy b.i.d. to a total of 69.6 Gy and concurrent chemotherapy in our 3 prospective studies were analyzed. The chemotherapy regimen was either (1) 50 mg each of carboplatin and etoposide (CE) given on RT days (163 patients) or (2) 30 mg of CE on RT days and 100 mg of CE on Saturdays and Sundays during the RT course (138 patients). An IFI of 4.5-5 h or 5.5-6 h had been nonrandomly assigned for each patient, and this interval was kept throughout the treatment. RESULTS: No difference was observed in treatment outcome due to the chemotherapy protocol, and the 2 groups were combined. Patients treated with the shorter IFI had a better local control rate (38% at 5 years) and survival rate (30% at 5 years) than those treated with the longer interval (23% and 14%, respectively; p < 0.001). However, female patients and those with a high Karnofsky performance status score (KPS), weight loss of < or =5% in the previous 6 months, or Stage IIIA disease had been more often treated with the shorter IFI, and these characteristics were associated with better treatment outcome. In multivariate analysis, only gender, KPS, and weight change proved to be significant prognostic factors influencing both local control and survival, and the effect of IFI was not significant. The incidence of Grade 4 acute esophagitis tended to be higher in the shorter interval group (p = 0.072), but there were no differences in the incidence of late or other acute RT-related toxicities between the 2 groups. CONCLUSIONS: The possible influence of the IFI on local control and survival could not be verified using multivariate analysis. To better understand the influence of the IFI, randomized studies with more patients and wider ranges of intervals (e.g., 5 h vs. 8 h) seem to be necessary. PMID- 11380215 TI - Clinical significance of serum pulmonary surfactant proteins a and d for the early detection of radiation pneumonitis. AB - PURPOSE: Radiation pneumonitis (RP) is one of the most serious complications for patients who receive thoracic irradiation. To avoid this, early diagnosis of radiation pneumonitis is extremely important. The purpose of the present study is to investigate whether serum pulmonary surfactant proteins A and D (SP-A and SP D, respectively) could be useful markers for RP. METHODS AND MATERIALS: Eighty six patients (lung cancer: 42 [primary: 39, metastatic: 3], breast cancer: 23, esophageal cancer: 21) who underwent radiation therapy were prospectively studied. Radiation doses ranged from 30-76 Gy (median, 58 Gy). Serum SP-A and SP D levels were evaluated sequentially by a sandwich enzyme-linked immunosorbent assay (ELISA) method before, during, and throughout the follow-up period until the development of symptomatic RP or until one year after completion of radiotherapy. Specificity of the ELISA results was confirmed by Western blot analysis. Patients symptomatic for RP were graded according to the Common Toxicity Criteria. RESULTS: RP occurred in 19 patients. Serum SP-D levels of patients with RP were sequentially higher than those in patients without RP. In the monitoring, serum SP-D levels at 50-60 Gy showed greater sensitivity and positive predictive values for RP detection (74% and 68%, respectively) than SP-A (26% and 21%, respectively). Western blot analysis showed that the development of RP was due to overproduction, but not proteolysis of surfactant proteins. CONCLUSION: We confirm that serum SP-A and SP-D monitoring is a practical and useful diagnostic method for the early detection of RP. PMID- 11380216 TI - Practice guideline on prophylactic cranial irradiation in small-cell lung cancer. AB - PURPOSE: To develop an evidence-based clinical practice guideline that would address the following questions: (a) What is the role of prophylactic cranial irradiation (PCI) in patients with limited or extensive stage small-cell lung cancer (SCLC) who have achieved complete remission in response to induction therapy (chemotherapy or chemoradiotherapy)? (b) What dose and fractionation schedules of PCI are optimal? (c) Does the use of PCI in patients with SCLC in complete remission affect quality of life? Survival, disease-free survival, quality of life, and adverse effects were the outcomes of interest. METHODS AND MATERIALS: A systematic review of the published literature was undertaken to provide the data for an evidence-based practice guideline. RESULTS: Six randomized controlled trials and one fully published individual patient data meta analysis were included in the systematic review of the evidence. For patients who have achieved complete response after induction therapy, there is evidence of a disease-free survival benefit (4 of 6 trials) and an overall survival benefit (meta-analysis). There is insufficient evidence to make a definitive recommendation with respect to dose. There is some indication that 30-36 Gy in 2 3 Gy per fraction, or a biologically equivalent dose, may produce a better outcome than a lower dose or less aggressive fractionation regimen. The schedule commonly used in Canada is 25 Gy in 10 fractions over 2 weeks. Data from further research, including a trial currently ongoing that compares 25 Gy in 10 fractions with 36 Gy in 18 fractions, will be required to determine optimal dose of PCI. There is insufficient evidence to make recommendations concerning the optimal timing of PCI in relation to the administration of chemotherapy. Lung DSG members generally felt that it should be given as soon as possible after completion of chemotherapy. There is evidence from trials with data for up to 2 years of follow up that prophylactic cranial irradiation does not produce significant late neurotoxicity. There is evidence from one trial that prophylactic cranial irradiation does not have a detrimental effect on quality of life in the first 12 months following the completion of therapy. There is insufficient evidence to comment on the long-term effects of prophylactic cranial irradiation on quality of life. CONCLUSION: For adult patients with limited or extensive SCLC who achieve a complete remission with induction therapy, PCI is recommended. PMID- 11380217 TI - Magnetic resonance spectroscopy of the malignant prostate gland after radiotherapy: a histopathologic study of diagnostic validity. AB - PURPOSE: Accurate spatial representation of tumor clearance after conformal radiotherapy is an endpoint of clinical importance. Magnetic resonance spectroscopy (MRS) can diagnose malignancy in the untreated prostate gland through measurements of cellular metabolites. In this study we sought to describe spectral metabolic changes in prostatic tissue after radiotherapy and validate a multivariate analytic strategy (based on MRS) that could identify viable tumor. METHODS AND MATERIALS: Transrectal ultrasound-guided prostate biopsies from 35 patients were obtained 18-36 months after external beam radiotherapy. One hundred sixteen tissue specimens were subjected to 1H MRS, submitted to histopathology, and analyzed for correlation with a multivariate strategy specifically developed for biomedical spectra. RESULTS: The sensitivity and specificity of MRS in identifying a malignant biopsy were 88.9% and 92% respectively, with an overall classification accuracy of 91.4%. The diagnostic spectral regions identified by our algorithm included those due to choline, creatine, glutamine, and lipid. Citrate, an important discriminating resonance in the untreated prostate gland, was invisible in all spectra, regardless of histology. CONCLUSIONS: Although the spectral features of prostate tissue markedly change after radiotherapy, MRS combined with multivariate methods of analysis can accurately identify histologically malignant biopsies. MRS shows promise as a modality that could integrate three-dimensional measures of tumor response. PMID- 11380218 TI - The 1989 patterns of care study for prostate cancer: five-year outcomes. AB - PURPOSE: Five-year results from the 1989 patterns of care study (PCS) for prostate cancer are now ready for analysis. The PCS was initiated to determine national averages for treatments and examine outcomes prospectively; the 1989 prostate study is the first to have collected pre- and post-treatment serum PSA data. METHODS AND MATERIALS: Six hundred patients treated with radiotherapy with curative intent for prostate cancer at 71 separate institutions in the year 1989 made up the study population. Three hundred ninety-one cases were fully analyzable. Pretreatment patient and tumor characteristics were as follows: of the 391 analyzable, 255 had pretreatment PSA values obtained, and 245 had a Gleason's sum (GS) reported. Three hundred fifty-eight were Caucasian, 24 African American, and 3 Hispanic (also 6 unknown). One hundred three patients had PSA < 10, 60 had PSA 10-19, and 92 presented with PSA >20. Ninety-seven patients were from Radiation Therapy Oncology Group (RTOG), Community Cancer Centers (CCC), or teaching institutions; 141 patients were from other hospital-based, nonteaching institutions; and 153 were from freestanding radiation oncology facilities. Seventy-one patients were T1, 203 T2, and 100 T3/4. Twenty-four out of 391 patients also received neoadjuvant hormone therapy. Survival curves were constructed using Kaplan-Meier methods, and differences between groups were tested for significance using the log-rank test. For cumulative incidence curves, Gray's test was used to investigate failure distributions between groups. The variables entering Cox model for multivariate analysis included age, race, T stage, pretreatment PSA, and GS. A patient was considered a PSA failure if the treating radiation oncologist reported it as such. RESULTS: With a median follow up of 5.7 years, the 5-year biochemical no evidence of disease (bNED) and overall survival were 56% and 79% respectively for Stage T1, 52% and 81% for T2, and 36% and 63% for Stages T3 and T4 combined. As expected, higher pretreatment PSA, GS, and T stage were all prognostic of poorer outcome. On univariate analysis, bNED survival was adversely impacted by T stage (p = 0.009), pretreatment PSA (p = 0.0035), and by the GS (p = 0.0038). Cause-specific failure was significantly lower for higher T stage (p = 0.014), GS (p = 0.001), and also pretreatment PSA (p = 0.0004). Overall survival was significantly lower in patients with higher T stage (p = 0.047) or GS (p = 0.0191), but not pretreatment PSA (p = 0.284). On multivariate analysis, pretreatment PSA was found to be statistically significant in association with bNED survival, and GS was associated with overall survival, cause-specific survival, and distant metastasis. Few late complications were reported: 13/391 and 13/391 Grade 2-3 gastrointestinal (GI) and genitourinary (GU) complications respectively, with two patients having required surgery with or without a permanent colostomy. CONCLUSION: For a representative cross-section of institutions in the United States, radiotherapy achieved high rates of bNED and CSS in selected groups of prostate cancer patients. When studied retrospectively, increased pretreatment PSA was a strong predictor of both biochemical failure and death due to prostate cancer. New strategies for patients with high-stage, high-grade tumors and/or pretreatment PSA >20 deserve testing. PMID- 11380219 TI - Defining the risk of developing grade 2 proctitis following 125I prostate brachytherapy using a rectal dose-volume histogram analysis. AB - OBJECTIVE: To determine the rectal tolerance for developing Grade 2 radiation proctitis after 125I prostate implantation based on the rectal dose-volume histogram. METHODS AND MATERIALS: Two hundred twelve patients with T1-T2 prostate cancer underwent 125I implantation without external beam irradiation. One month after the procedure, all patients underwent CT-based postimplant dosimetry (3-mm abutting slices). The rectal volumes, defined by an inner and outer wall, were determined from 9 mm above the seminal vesicles to 9 mm below the prostate apex. All doses were calculated by TG43 formalism. The prostate prescription dose was 160 Gy. A dose response analysis was undertaken for volumes of rectal tissue receiving a given dose. Dose levels examined were 80 Gy, 100 Gy, 120 Gy, 140 Gy, 160 Gy, 180 Gy, 200 Gy, 220 Gy, and 240 Gy. Grade 2 proctitis was defined as rectal bleeding occurring at least once a week for a minimum period of one month. The risk of proctitis was calculated using actuarial methods. For each dose level, a critical volume cutpoint was chosen to define a low and high volume group of patients. The cutpoint was determined based on two goals: minimizing thep value and finding a < or =5% risk of proctitis in the low volume group. Patients were followed from 12 to 61 months (median: 28 months) after implantation. RESULTS: Twenty-two patients developed Grade 2 proctitis: 14% within the first year, 72% between the first and second year, and 14% during the third year after the implant date. After the third year postimplantation, no cases of proctitis were reported. Proctitis was found to be significantly volume dependent for a given dose. The prescription dose (160 Gy) delivered to < or =1.3 cc of rectal tissue resulted in a 5% rate of proctitis at 5 years vs. 18% for volumes >1.3 cc (p = 0.001). Similar results were found for all doses examined. As the rectal volume receiving the prescription dose (160 Gy) increased, so did the proctitis rate: 0% for < or =0.8 cc, 7% for >0.8-1.3 cc, 8% for >1.3-1.8 cc, 24% for >1.8-2.3 cc, and 25.5% for >2.3cc (p = 0.002). CONCLUSIONS: Rectal dose volume histogram analysis is a practical and predictive method of assessing the risk of developing Grade 2 proctitis after 125I prostate implantation. Delivered dose should be kept below defined rectal volume thresholds to minimize this risk. This information can allow one to decrease rectal morbidity by modifying prostate implant technique. PMID- 11380220 TI - Mucositis reduction by selective elimination of oral flora in irradiated cancers of the head and neck: a placebo-controlled double-blind randomized study. AB - PURPOSE: The aim of the study was to test the hypothesis that aerobic Gram negative bacteria (AGNB) play a crucial role in the pathogenesis of radiation induced mucositis; consequently, selective elimination of these bacteria from the oral flora should result in a reduction of the mucositis. METHODS AND MATERIALS: Head-and-neck cancer patients, when scheduled for treatment by external beam radiation therapy (EBRT), were randomized for prophylactic treatment with an oral paste containing either a placebo or a combination of the antibiotics polymyxin E, tobramycin, and amphotericin B (PTA group). Weekly, the objective and subjective mucositis scores and microbiologic counts of the oral flora were noted. The primary study endpoint was the mucositis grade after 3 weeks of EBRT. RESULTS: Seventy-seven patients were evaluable. No statistically significant difference for the objective and subjective mucositis scores was observed between the two study arms (p = 0.33). The percentage of patients with positive cultures of AGNB was significantly reduced in the PTA group (p = 0.01). However, complete eradication of AGNB was not achieved. CONCLUSIONS: Selective elimination of AGNB of the oral flora did not result in a reduction of radiation-induced mucositis and therefore does not support the hypothesis that these bacteria play a crucial role in the pathogenesis of mucositis. PMID- 11380221 TI - Acupuncture for pilocarpine-resistant xerostomia following radiotherapy for head and neck malignancies. AB - OBJECTIVE: Xerostomia is a frequent and potentially debilitating toxicity of radiotherapy (XRT) for cancers of the head and neck. This report describes the use of acupuncture as palliation for such patients. METHODS AND MATERIALS: Eighteen patients with xerostomia refractory to pilocarpine therapy after XRT for head and neck malignancy were offered acupuncture as palliation. All patients are without evidence of cancer recurrence at the primary site. Acupuncture was provided to three auricular points and one digital point bilaterally, with electrostimulation used variably. The Xerostomia Inventory (XI) was administered retrospectively to provide an objective measure of efficacy. RESULTS: Acupuncture contributed to relief from xerostomia to varying degrees. Palliative effect as measured by the XI varied from nil to robust (pre- minus post- therapy values of over 20 points). Nine patients had benefit of over 10 points on the XI. CONCLUSIONS: Acupuncture reduces xerostomia in some patients who are otherwise refractory to best current management. PMID- 11380222 TI - Squamous cell carcinomas of the soft palate treated with radiation therapy alone or followed by planned neck dissection. AB - PURPOSE: The present study presents the experience at the University of Florida with treatment of unselected patients with carcinomas of the soft palate with radiation therapy (RT) alone or followed by planned neck dissection. METHODS AND MATERIALS: One hundred seven patients treated with curative intent with RT alone or followed by neck dissection from 1965 to 1996 were included in the study. All patients had follow-up for at least 2 years. No patients were lost to follow-up. RESULTS: Local control rates at 5 years were 86% for T1, 91% for T2, 67% for T3, and 36% for T4 carcinomas. T-stage and overall treatment time significantly affected local control in multivariate analysis. Nodal control rates at 5 years were 86% for NO, 76% for N1, 61% for N2, and 67% for N3 carcinomas. Overall treatment time and planned neck dissection significantly affected nodal control in multivariate analysis. Ultimate local-regional control rates at 5 years were 90% for Stage I, 92% for Stage II, 84% for Stage III, and 60% for Stage IV disease. Overall treatment time and planned neck dissection significantly affected ultimate local-regional control in multivariate analysis. The overall survival rate at 5 years was 42% for all patients. Overall stage, overall treatment time, and planned neck dissection significantly affected overall survival in multivariate analysis. The cause-specific survival rate at 5 years was 70% for all patients. Overall treatment time and planned neck dissection significantly affected cause-specific survival in multivariate analysis. Three patients sustained severe postoperative complications and 3 patients sustained severe late complications. Sixteen patients had synchronous and 14 patients had metachronous carcinomas of the head and neck mucosal sites. CONCLUSION: For limited carcinomas of the soft palate, RT (alone or followed by planned neck dissection) results in relatively high local-regional control and survival rates. For advanced carcinomas of the soft palate, local-regional control and survival rates are relatively low and local-regional recurrence rates are substantial. Advanced carcinomas of the soft palate may be better treated with RT and concomitant chemotherapy. PMID- 11380223 TI - The role of postradiotherapy neck dissection in supraglottic carcinoma. AB - PURPOSE: To evaluate our policy of performing neck dissection based on regional response after definitive radiotherapy in patients with supraglottic carcinoma and to identify the prognostic factors in this group of patients. METHODS AND MATERIALS: Between 1970 and 1995, 121 patients with node-positive squamous cell carcinoma of the supraglottic larynx were treated with definitive radiotherapy. Sixty-nine percent of patients presented with 1997 AJCC Stage IV disease. The N stage distribution was N1, 49; N2, 62; and N3, 10. The median size of the lymph nodes was 3 cm (range, 0.5-8 cm). Forty-five patients received once-a-day treatment with a median total dose of 65 Gy (range, 58.0-70.8 Gy) in 1.8-2.0 Gy per fraction over 48 days, and 76 patients received split-course accelerated hyperfractionation with a median total dose of 67.2 Gy (range, 63.2-73.6 Gy) in 1.6 Gy twice a day over 43 days. Patients whose lymph nodes were not clinically detectable at 4-6 weeks after the completion of radiotherapy (complete response) were followed without any neck dissection. Patients with persistent neck adenopathy (partial response) underwent neck dissection whenever possible. Mean follow-up of the living patients was 6.5 years. RESULTS: Regional response was related to the size of lymph nodes at presentation. Eighty-seven percent of patients with nodal size of 3 cm or less had a complete response, whereas 43% of patients with nodal size greater than 3 cm had a partial response. The rate of regional control at 3 years for all patients in the study was 66%. The 3-year ultimate regional control rate after salvage neck dissection was 75%. A relapse in both the primary and regional sites was the most common pattern of relapse, accounting for 39% of all the failures. Local failure was associated with subsequent regional relapse with a relative risk of 4.3. For patients with complete response in whom postradiotherapy neck dissection was withheld, the regional control rates were 75% and 86% for N1 and N2, respectively. The rate of isolated regional relapse in this group of patients was 7.5%. In multivariate analysis, significant favorable factors predictive for regional control were female gender, accelerated hyperfractionation, and complete response; whereas factors predictive for overall survival were Karnofsky Performance Scale score and regional response. The rate of Radiation Therapy Oncology Group (RTOG) Grade 2 or 3 neck fibrosis was 17% and 23% for patients with and without postradiotherapy neck dissection, respectively. CONCLUSION: Isolated regional relapse is not common in patients with supraglottic carcinoma when a complete response is achieved at 4-6 weeks after definitive radiotherapy and postradiotherapy neck dissection is not performed. Female gender, accelerated hyperfractionation, and complete response are favorable predictors of regional control. PMID- 11380224 TI - Conformal re-irradiation of recurrent and new primary head-and-neck cancer. AB - PURPOSE: To review the outcome of head-and-neck cancer patients re-irradiated using conformal radiation. PATIENTS AND METHODS: From 1983 to 1999, 60 patients with recurrent or new primary head-and-neck cancer received re-irradiation at the University of Michigan. Twenty patients were excluded due to the planned cumulative radiation dose being less than 100 Gy (18) and absence of prior radiation details (2), leaving 40 patients. Thirty-five patients were re irradiated for unresectable disease, while 4 patients received adjuvant re irradiation for high-risk disease. Thirty-eight patients had recurrences from previously treated cancer (19 regional, 14 local, 5 regional and local), and 2 patients had new primary tumors. The median time from the first course of radiation to re-irradiation was 21 months. Thirty-one patients (78%) were re irradiated with curative intent, whereas 9 were treated with palliative intent. Re-irradiation was delivered using conformal techniques in the majority of patients and with concurrent chemotherapy in 14 patients. The median re irradiation dose was 60 Gy. The median cumulative dose received was 121 Gy. Five patients (13%) did not complete their prescribed course of re-irradiation. RESULTS: The median survival following completion of re-irradiation was 12.5 months. The 1- and 2-year actuarial survival rates were 51.1% and 32.6%, respectively. On multivariate analysis, palliative intent of treatment, tumor bulk, and tumor site other than nasopharynx or larynx were associated with worse survival. The patients treated for unresectable disease did no worse than those treated adjuvantly. The median times to relapse-free survival, local-regional recurrence (LRR)-free survival, and ultimate LRR-free survival (allowing for surgical salvage) were 3.9 months, 7.8 months, and 8.7 months, respectively. Seven patients (18%) are presently alive with no evidence of disease, with a median follow-up of 49.9 months (range 3.3-78.9). Severe radiation-induced complications were seen in 7 patients (18%). Two other patients developed orocutaneous fistulas in the presence of tumor recurrence. Moderate fibrosis and trismus were common. CONCLUSION: Despite the use of conformal techniques, the prognosis of patients treated with re-irradiation is poor, and complications are not infrequent. A subset of patients is salvageable, and high-dose re-irradiation should be considered in selected patients. PMID- 11380225 TI - Isolated late local recurrences with high mitotic count and early local recurrences following breast-conserving therapy are associated with increased risk on distant metastasis. AB - PURPOSE: Local recurrence (LR) after breast-conserving therapy (BCT) is associated with an increased risk for the development of distant metastasis. We studied risk factors for distant metastasis risk (DMR) and poor prognosis within a group of patients with LR as first event. PATIENTS AND METHODS: From a cohort of 1481 breast carcinomas treated with BCT in the period 1980-1994, a total of 68 pT1-3 N0-1 patients developed LR as first event. We have studied risk factors for the development of distant metastasis within this group of patients with LR. In addition to clinical factors (age at BCT and LR, mode of detection, location of LR, and treatment of LR), the histology slides of the primary and the recurrent tumor were reviewed. Immunohistochemical staining was performed for the following proteins: bcl-2, cyclin D1, E-cadherin, EGF receptor, ER, PR, Ki-67, c-erbB 2/neu, and p53. Statistical analyses were performed using conditional logistic regression. RESULTS: At a median follow-up after LR of 5.6 years, the 5-year DMR was 53%. In univariate analysis, none of the factors of the primary tumor was found to be associated with DMR after LR. Of the recurrent tumor the following factors were found to be risk factors for high DMR after LR: interval between treatment of the primary tumor and LR at 2 years or less (relative risk, 2.38; 95% confidence interval, 1.22-4.76; p = 0.008) and high mitotic count (relative risk, 2.51; 95% confidence interval, 1.03-6.15; p = 0.04). All patients with noninvasive recurrent tumor were alive at the time of analysis. Patients with an interval of greater than 2 years and a recurrent tumor with high mitotic count were found to have an equally poor prognosis compared to patients with LRs detected after a short interval. CONCLUSION: LR after BCT is associated with higher DMR and poor prognosis. Patients with LR within 2 years after BCT are especially at high risk. Late recurrences with high mitotic count have the same poor prognosis as early recurrences. For these patients, systemic treatment at time of the detection of LR should be considered. PMID- 11380226 TI - Recursive partitioning analysis of locoregional recurrence patterns following mastectomy: implications for adjuvant irradiation. AB - PURPOSE: Postmastectomy irradiation improves overall survival for breast cancer patients at high risk for locoregional recurrence (LRR). The objective of this study was to use recursive partitioning analysis (RPA) to define patient subgroups at high risk for LRR following mastectomy. PATIENTS AND METHODS: A cohort of 1031 patients treated on prospective trials with mastectomy and doxorubicin-based chemotherapy without irradiation was analyzed. The variables considered in the RPA were tumor size, number of involved nodes, number of nodes examined, and percentage of nodes involved (nodes involved/nodes examined). The endpoint was LRR +/- distant metastasis. Only patients with complete data were analyzed (n = 913). Median follow-up was 8 years (range, 0.7-22 years). RESULTS: Involvement of 20% or more of the lymph nodes examined was the most significant variable predicting LRR. Three risk categories were defined. Patients with 20% or more involved nodes and tumors of 3.5 cm or more were at greatest risk for LRR (41% at 8 years). An intermediate-risk group included patients with 20% or more involved nodes and tumors of less than 3.5 cm as well as those with less than 20% involved nodes and tumor size of 5 cm or greater (18% at 8 years). Patients with less than 20% involved nodes and tumor size of less than 5 cm were at lowest risk for LRR (10% at 8 years). CONCLUSION: Tumor size and extent of nodal involvement play interrelated roles in predicting LRR following mastectomy and systemic therapy. Patients with 20% or greater involved nodes and those with less than 20% nodes and tumors of 5.0 cm or greater are at significant risk of LRR and should be considered for postoperative irradiation. PMID- 11380227 TI - Can 8-oxo-dG be used as a predictor for individual radiosensitivity? AB - PURPOSE: To develop predictive tests for individual radiosensitivity of tumor patients. METHODS AND MATERIALS: Acute skin reactions were clinically scored among 40 women after 46 Gy, given with 2 Gy fractions to breast and regional lymph nodes, adjuvant after surgery. The acute skin reactions were compared to the excretion of 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dG) in urine, determined by high-performance liquid chromatography (HPLC) with electrochemical detector. Specimens of urine were collected before and during postoperative radiation treatment at given intervals. We compared a group of 9 patients with the most pronounced skin reactions with another group of 8 patients with almost no skin reactions at 46 Gy. RESULTS: The level of 8-oxo-dG excreted in urine during 8 h was measured. After normalizing the excretion to irradiated volumes, dose per volume and excretion before irradiation, the 8-oxo-dG level in urine was significantly (p < 0.001) lower for the patients with pronounced skin reactions as compared to patients with minor skin reactions, at an accumulated dose of 12 Gy. In addition, the background level of 8-oxo-dG excreted before treatment started, was significantly (p = 0.043) lower for patients with minor skin reactions as compared to patients with pronounced skin reactions. The background level of 8-oxo-dG was corrected for body weight and normalized to BMI. CONCLUSION: We suggest that the excretion of 8-oxo-dG into urine of breast cancer patients is a possible marker for acute radiosensitivity. PMID- 11380228 TI - Pulmonary sequelae of treatment for breast cancer: a prospective study. AB - PURPOSE: To prospectively study the effects of loco-regional radiotherapy in women with breast cancer. METHODS AND MATERIALS: Thirty consecutive patients with breast resection underwent clinical, lung function, radiographic, and thoracic high-resolution computed tomography evaluation before and at 1, 3, 6, and 12 months after adjuvant radiotherapy. Chemotherapy was also administered to 15 patients. RESULTS: Nineteen patients reported mild respiratory symptoms at 1 month, which resolved completely at 6 months after radiotherapy. Opacities were present on 80% of chest radiographs and in all patients on high-resolution computed tomography by 3 months. These opacities became compact and persisted on high-resolution computed tomography at 12 months. Lung function indices, including FEV1, FVC, TLC, and DLCO, progressively declined after radiotherapy, and was irreversible at 12 months (p < 0.05). Patients who received chemotherapy did not have significantly different lung function indices compared with their counterparts at all time points (p > 0.05). CONCLUSIONS: Our results have shown that adjuvant loco-regional radiotherapy, a common practice in breast cancer treatment, is associated with irreversible reduction in lung function parameters. These changes are accompanied by radiological evidence of persistent lung injury. Further studies should be performed to evaluate the incidence and long-term pulmonary sequelae of current treatment for breast cancer. PMID- 11380229 TI - A practical and easy method to locate the first three internal mammary lymph node bearing areas. AB - PURPOSE: To assess the distance from a clinically recognized anatomic landmark to the different costocondral interspaces in female patients to facilitate the design of radiation fields intended to include specific internal mammary nodal areas. METHODS AND MATERIALS: The distance from the suprasternal notch (SSN) to the caudal portion of the first through fourth interspace was measured on a computer display of the chest skeleton of 65 female patients with left-sided breast cancer. The relationship between these distances and bone size (sternal length and standing height) was assessed via linear regression. In 21 of the 65 patients where myocardial perfusion imaging of the heart was available, the relationship between the location of the 3rd costochondral interspace and the left ventricle was assessed. RESULTS: In 90% of patients (59/65), the first, second, third, and fourth interspace were within 5, 8.5, 11, and 14 cm of the SSN, respectively. The SSN-interspace distances did not correlate well with sternal length (r = 0.28) or standing height (r = 0.31). In 20 of 21 patients (95%), the third interspace "shadowed" the cephalad aspect of the left heart ventricle. Median "shadowing" was 3 cm (range 0.5-6 cm). CONCLUSION: The caudal portion of the third costochondral interspace is < or = 11 cm caudal to the SSN in 90% of patients. These measurements can be used to clinically design radiation therapy fields intended to treat the upper three interspaces. The distance from the SSN to the 1st through 4th interspaces is not related to sternal length or to standing height. In patients with left-sided breast cancer, radiation treatment fields designed to include the internal mammary lymph nodes in the upper three interspaces may incidentally include a portion of the heart. PMID- 11380230 TI - Combined chemo- and radiotherapy vs. radiotherapy alone in the treatment of primary, nonresectable adenocarcinoma of the rectum. AB - PURPOSE: In a randomized study in primarily inextirpable rectal cancer, conventional radiotherapy to reduce the tumor mass was compared with combined chemotherapy and radiotherapy. METHODS AND MATERIALS: The combined treatment (CRT) was given every other week, four times, during a 7-week period. The drugs used were methotrexate, 5-fluorouracil in bolus injection followed by continuous infusion and leucovorin rescue. Radiotherapy (RT) was given simultaneously with five 2-Gy fractions in 3 days to a dose of 10 Gy to a total dose in the four courses of 40 Gy. This regimen was compared with radiotherapy in 2-Gy fractions to a total dose of 46 Gy in the radiotherapy group. Surgery was performed 3-4 weeks after finished treatment. Seventy patients were included between November 1988 and August 1996; 36 patients were allocated to RT and 34 to CRT. RESULTS: Twenty-five (74%) of the patients in the CRT group underwent a locally radical resection with 20 (59%) patients without any known metastases. The corresponding figures in the RT group were 23 (64%) and 18 (50%), respectively. Among the patients who underwent any tumor resection, 5/29 (17%) in the CRT group and 12/27 (44%, p = 0.05) in the RT group have had a local recurrence. After a locally radical resection, the corresponding figures are 4% and 35% (p = 0.02), respectively. Local disease-free survival was significantly superior in the CRT group (66% at 5 years) compared with the RT group (38%, p = 0.03 log-rank test). Five-year survival was 29% (9 patients) in the CRT group and 18% (6 patients) in the RT group, a nonsignificant difference (p = 0.3). Five patients in the RT group did not complete planned treatment, mainly due to the appearance of metastatic disease. In this group toxicity was usually of Grade 0-1. In the experimental group, the toxicity usually was Grade 2 or higher, and 6 patients did not manage to fulfill the planned treatment due to toxicity. CONCLUSION: In this study, with fewer included patients than intended, resectability rates were high in both groups. The addition of chemotherapy to radiotherapy significantly improved local control rates, but no statistically significant difference was found in survival between the groups. The acute toxicity after CRT was higher than after RT alone, but manageable. PMID- 11380231 TI - Preoperative chemoradiotherapy with oral doxifluridine plus low-dose oral leucovorin in unresectable primary rectal cancer. AB - PURPOSE: The use of oral chemotherapeutic agents in chemoradiotherapy provides several advantages. Doxifluridine, an oral 5-FU prodrug, has been shown to be effective in colorectal cancer. We attempted a Phase II trial of preoperative chemoradiotherapy with doxifluridine plus a low-dose oral leucovorin in unresectable primary rectal cancer patients. In this study, toxicity and efficacy were evaluated. METHODS AND MATERIALS: There were 23 patients with primary unresectable rectal cancer in this trial, 21 of whom were available for analysis. The patients were treated with oral doxifluridine (900 mg/day) plus oral leucovorin (30 mg/day) from days 1 to 35, and pelvic radiation of 45 Gy over 5 weeks. Surgical resection was performed 5-6 weeks after the treatment. RESULTS: Acute toxicity involved thrombocytopenia, nausea/vomiting, diarrhea, and skin reaction. All were in Grade 1/2, except diarrhea, which was not only the most frequent (7 patients, 33.3%), but also the only toxicity of Grade 3 (2 patients). The clinical tumor response was shown in 5 patients (23.8%) as a complete response and 13 patients (61.9%) as a partial response. A complete resection with negative resection margin was done in 18 patients (85.7%), in 2 of whom a pathologic complete response was shown (9.5%). The overall downstaging rate in the T- and N-stage groupings was 71.4% (15 patients). CONCLUSION: This study demonstrated the efficacy and low toxicity of chemoradiotherapy with doxifluridine. Currently, a Phase III randomized trial of chemoradiotherapy is ongoing at our institute to compare the therapeutic efficacy of oral 5-FU with respect to i.v. 5-FU in locally advanced and unresectable rectal cancer. PMID- 11380232 TI - Extracorporeal irradiation for malignant bone tumors. AB - PURPOSE: Extracorporeal irradiation (ECI) has been used selectively in the management of primary malignant bone tumors since 1996. We report our techniques for ECI and the short-term oncologic and orthopedic outcomes. METHODS AND MATERIALS: Sixteen patients with primary malignant bone tumors were treated with ECI from 1996 to 2000. The median age was 14 years. The histologic diagnoses were Ewing's sarcoma (11), osteosarcoma (4) and chondrosarcoma (1). The treated sites were femur (7), tibia (4), humerus (2), ilium (2), and sacrum (1). Following induction chemotherapy in Ewing's sarcomas and osteosarcoma, en bloc resection of the tumor and tumor-bearing bone was performed. A single dose of 50 Gy was delivered to the bone extracorporeally using either a linear accelerator (9 cases) or a blood product irradiator (7 cases). The orthopedic outcome was recorded using a standard functional scale. RESULTS: At a median follow-up of 19.5 months, there were no cases of local recurrence or graft failure. One patient required amputation due to chronic osteomyelitis. For the 10 patients with follow-up greater than 18 months, the functional outcomes were graded good to excellent. CONCLUSION: The short-term oncologic and orthopedic results are encouraging and suggest that ECI provides a good alternative for reconstruction in limb conservative surgery in selected patients. This technique should only be used in a multidisciplinary setting, where careful follow-up is available to assess the long-term outcomes. PMID- 11380233 TI - Fast neutron radiotherapy for soft tissue and cartilaginous sarcomas at high risk for local recurrence. AB - PURPOSE: The practice policy at the University of Washington has been to employ fast neutron radiotherapy for soft tissue sarcoma lesions with prognostic features predictive for poor local control. These include gross residual disease/inoperable disease, recurrent disease, and contaminated surgical margins. Cartilaginous sarcomas have also been included in this high-risk group. This report updates and expands our previously described experience with this approach. METHODS AND MATERIALS: Eighty-nine soft tissue sarcoma lesions in 72 patients were treated with neutron radiotherapy in our department between 1984 and 1996. Six patients, each with solitary lesions, were excluded from analysis due to lack of follow-up. Seventy-three percent were treated with fast neutron radiation alone, the rest with a combination of neutrons and photons. Median neutron dose was 18.3 nGy (range 4.8-22). Forty-two patients with solitary lesions were treated with curative intent. Thirty-one patients (including 7 previously treated with neutrons) with 41 lesions were treated with the goal of local palliation. Tumors were predominantly located in the extremity and torso. Thirty of 35 (85%) of curative group patients treated postoperatively had close or positive surgical margins. Thirty-four (82%) lesions treated for palliation were unresectable. Thirty-five patients (53%) were treated at the time of recurrence. Median tumor size at initial presentation was 8.0 cm (range 0.6-29), median treated gross disease size was 5.0 cm (range 1-22), and 46/69 evaluable lesions (67%) were judged to be of intermediate to high histologic grade. Fourteen patients (21%) had chondrosarcomas. RESULTS: Median follow-up was 6 months (range 2-47) and 38 months (range 2-175) for the palliative and curative groups, respectively. Kaplan-Meier estimates were obtained for probability of local relapse-free survival (68%), distant disease-free survival (59%), cause specific survival (68%), and overall survival (66%) at 4 years for the curatively treated group. For the palliatively treated group, estimated local relapse-free survival at 1 year was 62%. Log-rank analysis of the curative group revealed recurrent disease to be the only risk factor predictive for significantly worse local and distant disease-free survival. Intermediate-/high-grade histology was predictive for inferior overall survival. Effective clinical response was documented for 21/27 (78%) lesions treated palliatively. Ten patients (15%) experienced serious chronic radiation-related complications. All of these patients had clinical situations requiring delivery of high neutron doses and/or large radiotherapy fields. CONCLUSION: Fast neutron radiotherapy is locally effective for soft tissue and cartilaginous sarcomas having well-recognized high risk features. Results in the palliative setting appear to be particularly encouraging, with neutrons frequently providing significant symptomatic response for gross disease, with minimal serious chronic sequelae. Fast neutron radiotherapy should be considered in patients at high risk for local recurrence in both the curative and palliative settings. PMID- 11380234 TI - CHOD/BVAM regimen plus radiotherapy in patients with primary CNS non-Hodgkin's lymphoma. AB - PURPOSE: To assess the efficacy and toxicity, including long-term neurotoxicity, of combined therapy with the CHOD/BVAM regimen given before cranial radiotherapy in the treatment of primary CNS lymphoma (PCNSL). METHODS AND MATERIALS: Thirty one consecutive patients with PCNSL were treated with one cycle of cyclophosphamide, doxorubicin, vincristine, and dexamethasone (CHOD) and two of carmustine (BCNU), vincristine, cytosine arabinoside, and methotrexate (BVAM), followed by cranial radiotherapy (45 Gy whole brain plus a 10-Gy boost for single lesions). The median age was 59 years (range 21-70) and 39% had poor performance status. The median follow-up of patients was 4.1 years (range 2.7-9.0). RESULTS: Twenty-one patients had no PCNSL at the end of treatment. The 5-year actuarial probability of survival was 31% (95% confidence interval [CI]: 11%-57%), with a median survival of 38 months. Patients < 60 years had a survival significantly longer than those > or = 60 years (4-year survival: 58% (95% CI: 34-82%) vs. 29% (95% CI: 5-53%), respectively; p = 0.04). Two patients died during chemotherapy from pulmonary embolism and bronchopneumonia, respectively, with no evidence of PCNSL at the autopsy. Dementia probably related to treatment occurred in 5 (62%) of the 8 patients 60 years and older, and 4 of them died without evidence of relapse of PCNSL. Dementia correlated with developing brain atrophy and leuco encephalopathy on serial CT or MR scans. CONCLUSION: This regimen can be given with the planned dose intensity to patients aged less than 70 years, and produces better survival than that reported with radiotherapy alone; however, dementia occurs in the majority of patients aged 60 years of age or more. PMID- 11380235 TI - Effect of fractionated regional external beam radiotherapy on peripheral blood cell count. AB - PURPOSE: The purpose of this study was to assess the need for obtaining weekly complete blood count (CBC) values and to identify the pattern of changes in CBC during regional conventional fractionated radiotherapy. METHODS AND MATERIALS: A retrospective analysis of CBC data on 299 adult cancer patients who received definitive conventional radiotherapy to head and neck (n = 95), chest (n = 96), and pelvis (n = 108) was performed. Temporal patterns and magnitude of change in white blood cells, neutrophils, lymphocytes, and platelets during radiotherapy were examined. RESULTS: There were statistically significant declines in all counts, albeit not clinically significant. Notable differences between disease sites were found. The greatest weekly interval change in counts occurred during the first week of radiotherapy for all groups of patients. The mean WBC nadir values during treatment were 5.8 for head & neck, 6.8 for chest, and 5.4 for pelvis. The nadirs for all counts occurred toward the middle-to-end of radiotherapy. Lymphocytes were found to be more sensitive to radiotherapy than other leukocyte subcomponents. CONCLUSION: Our study suggests that weekly CBC monitoring is not necessary for all patients undergoing standard fractionated radiotherapy. Baseline blood counts may be used to determine an optimal schedule for monitoring CBCs in patients receiving conventional radiation alone. Reduced monitoring of CBC may result in significant financial savings. PMID- 11380236 TI - Selective radioprotection of hepatocytes by systemic and portal vein infusions of amifostine in a rat liver tumor model. AB - PURPOSE: The tolerance of the liver to radiation is too low to permit an effective dose to be delivered to patients who have diffuse intrahepatic cancer. In this study we evaluated whether systemic or portal venous administration of the aminothiol compound, amifostine, could protect the normal liver from the effects of ionizing radiation without compromising tumor cell kill in a rat liver tumor model. METHODS AND MATERIALS: Rats implanted with liver tumors were infused with 200 mg/kg amifostine over 15 min via the femoral or portal vein. The livers were irradiated with a single 6-Gy fraction 15-20 min after the termination of amifostine infusion. Protection of the liver was assessed by an in vitro hepatocyte micronucleus assay and tumor protection by an in vivo-in vitro clonogenic survival assay. Tissue levels of the active metabolite, free WR-1065, were determined in the tumor and in the normal liver using a specific HPLC assay with electrochemical detection. RESULTS: After a 6-Gy fraction, the frequency of hepatocyte micronuclei after administration of saline, systemic amifostine, and portal venous amifostine was 18.7+/-1%, 6.8+/-1%, and 9.9+/-2%, respectively, corresponding to a radiation equivalent effect of 6 Gy +/- 0.5, 1.8 Gy +/- 0.3, and 2.5 Gy +/- 1.3, respectively. Both amifostine conditions showed considerably less radiation effect than saline-treated controls (p < 0.01); the two amifostine conditions did not differ (p = 0.3). The surviving fraction of tumor cells was not affected by amifostine treatment and was 0.03+/-0.02 and 0.05+/-0.03 for systemic and portal venous delivery and 0.06+/-0.02 for control animals (ANOVA analysis showed no significant difference of the means p = 0.34). Portal venous delivery produced significantly less WR-1065 in the tumor compared to systemic administration (54 microM +/- 36 vs. 343 microM +/- 88, respectively, p = 0.03). CONCLUSIONS: Both systemic and portal venous administration of amifostine effectively protect hepatocytes from ionizing radiation without compromising tumor cell kill in a clinically relevant animal model. These findings suggest that amifostine may be a selective normal tissue radioprotectant in liver cancer and that regional/portal infusions may be preferable to systemic dosing. PMID- 11380237 TI - Amifostine before fractionated irradiation protects bone growth in rats better than fractionation alone. AB - The aim of this study was to determine the independent and combined effects of 100 mg/kg and 200 mg/kg doses of the radioprotectant amifostine and radiotherapy dose fractionation in preserving the integrity of or minimizing damage to the physis during high-dose radiation exposure in an animal model. Thirty-six weanling four-week-old male Sprague-Dawley rats were randomized into six study groups of six animals each. The distal femur and proximal tibia in the right leg of each animal was exposed to X-irradiation, with the contralateral left leg serving as the nonirradiated control. Three groups received a single 25 Gy radiotherapy dose: one group alone, a second group preceded by 100 mg/kg amifostine, and a third preceded by 200 mg/kg amifostine. Three groups received a total of 25 Gy in three equal fractions: one group alone, a second group preceded by 100 mg/kg amifostine, and a third preceded by 200 mg/kg amifostine. Fractionation of the 25 Gy radiation dose reduced the mean percent overall limb growth loss to 44.8%, a statistically significant reduction compared to a mean 58.8% reduced growth with the single 25 Gy dose. Addition of amifostine at 100 and 200 mg/kg before each of the three fractions of radiotherapy further decreased the mean percent overall limb growth loss to 35.2% and 28.5%, respectively, both statistically significant reductions beyond that achieved by fractionation alone. PMID- 11380238 TI - Inhibition of vascular cell growth by X-ray irradiation: comparison with gamma radiation and mechanism of action. AB - PURPOSE: Catheter-based delivery of gamma and beta radiation effectively inhibits restenosis. Major disadvantages of these radioisotopes include continuous emission; excessive depth of penetration, creating safety hazards (gamma); and inadequate penetration, limiting effectiveness (beta). Low-voltage X-rays have a distinct potential advantage, because the source is active only when current is applied, and depth of penetration is voltage dependent. This study was performed to determine if low-voltage X-rays inhibit smooth muscle and adventitial cell growth in vitro and to determine the molecular mechanisms involved in this cellular response. METHODS AND RESULTS: Vascular cells in culture were exposed to low-voltage X-ray radiation and analyzed for their subsequent ability to proliferate. X-ray irradiation caused a dose-dependent inhibition in proliferation, similar to the effect seen with equivalent doses of gamma radiation. The radiation-induced inhibition of proliferation did not appear to be related to apoptosis, but rather to delayed progression through the cell cycle, because a 65% increase in the proportion of cells in S phase was seen 24-96 h after X-ray exposure compared to control. Expression of p53, a cell cycle transcriptional activator, and p21, a cell cycle inhibitor, were significantly elevated after exposure to low-voltage X-rays, providing a potential mechanism for this delay. CONCLUSIONS: Low-voltage X-rays can effectively inhibit proliferation of vascular smooth muscle and adventitial cells. This inhibition is apparently due to a delay in progression through the cell cycle, which is mediated by increases in the levels of cell cycle inhibitors. PMID- 11380239 TI - Delayed re-endothelialization and T-cell infiltration following intracoronary radiation therapy in the porcine model. AB - PURPOSE: To evaluate the late induction of apoptosis following intracoronary radiation (IR) and the effects of IR on inflammatory cells. METHODS AND MATERIALS: Porcine coronaries were injured by balloon overstretch followed by either 0 or 15 Gy of 192Ir prescribed to 2 mm from the center of the source. Swine were euthanized at 3, 7, and 14 days posttreatment, and arteries were stained for markers of smooth muscle cells (SMCs alpha-actin), T cells (CD3), macrophages, endothelial cells, and apoptotic nuclei (terminal uridine nick end labeling, TUNEL). Intimal area (IA) and IA corrected for medial fracture length (IA/FL) were quantified by digital image analysis, which was also used to quantify the distribution of immunostain-positive cells in the adventitia, media, and neointima, respectively. RESULTS: IA/FL was significantly reduced following treatment with 15 Gy, in association with decreased SMC density. Following injury and IR, TUNEL- and CD3-positive cell density increased significantly, and density of macrophages was increased in the adventitia and neointima. Staining for endothelial cells revealed a delay of re-endothelialization after radiation treatment. CONCLUSION: Increased T-cell infiltration at the medial tear following IR, perhaps due to incomplete re-endothelialization, may indicate incomplete healing. The elevated apoptosis of these infiltrating T cells may indicate a mechanism for the resolution of inflammation. PMID- 11380240 TI - Ability to undergo apoptosis does not correlate with the intrinsic radiosensitivity (SF2) of human cervix tumor cell lines. AB - PURPOSE: To investigate the relationship between radiation-induced apoptosis and clonogenic cell kill in 9 cervical cancer cell lines. METHODS AND MATERIALS: Cells were irradiated with 0, 2, 8, and 30 Gy. The level of apoptosis was evaluated using flow cytometry (Annexin-V binding), light microscropy (morphology), gel electrophoresis (DNA ladder formation), and TUNEL assay. Cell survival was measured using a clonogenic assay. RESULTS: Of the 9 cervical carcinoma cell lines analyzed, 3 underwent radiation-induced apoptosis: CaSki, HT3, and 778. The levels of apoptosis, obtained 72 h after a dose of 30 Gy, were 49%, 28%, and 26%, respectively. All cell lines exhibited some level of background apoptosis measured by Annexin-V binding (mean = 2.6%+/-0.8; range, 0.2 6.9%) that correlated with the level of radiation-induced apoptosis (r = 0.92, p = 0.001). In 6 of the 9 lines, necrosis was the dominant form of cell death. A significant inverse relationship was found between the level of radiation-induced apoptosis and necrosis after 30 Gy (r = -0.87, p = 0.002). No relationship was found between radiation-induced apoptosis and intrinsic radiosensitivity measured, using a clonogenic assay, as surviving fraction at 2 Gy (SF2). CONCLUSION: Cervical carcinoma cells do not readily undergo radiation-induced apoptosis in vitro. There is no relationship between ability to undergo apoptosis and intrinsic radiosensitivity measured using a clonogenic assay. PMID- 11380241 TI - ATM protein expression correlates with radioresistance in primary glioblastoma cells in culture. AB - PURPOSE: Glioblastoma multiforme (GBM) is one of the malignancies most resistant to radiation therapy. In contrast, cells derived from individuals with ataxia telangiectasia (AT), possessing mutations in the ATM gene, demonstrate increased sensitivity to ionizing radiation. Using a collection of glioma specimens adapted to tissue culture and several established GBM cell lines, we investigated the relationship between ATM protein expression and radiosensitivity. The three aims of our study were to: (1) quantify ATM protein levels in cultured glioma cells; (2) measure the correlation between ATM protein levels and radiation sensitivity; and (3) examine the dependence of ATM on p53 status. METHODS AND MATERIALS: Glioma specimens were collected, catalogued, and adapted to grow in culture. Levels of ATM, p53, and p21 proteins were determined by Western blot. Radiation sensitivities were determined by clonogenic assays. p53 mutation status was determined by DNA sequencing. Correlations were identified by linear regression analysis. RESULTS: ATM protein levels were variable in the primary gliomas. Glioma cell lines demonstrated significantly lower levels of ATM protein. Clonogenic assays of cell strains and cell lines yielded survival fractions (SF2s) consistent with the radioresistant behavior of GBM tumors in vivo. Regression analysis revealed a high correlation between ATM protein levels and SF2 for primary glioma cell strains, but not for established GBM cell lines. p53 status failed to predict radiosensitivity. CONCLUSION: We have demonstrated that while our collection of low passage cell cultures depends on ATM for their resistance to IR, established cell lines may acquire adaptive characteristics which downplay the role of the ATM gene product in vitro. Therefore, attenuating ATM gene expression may be a successful strategy in the treatment of GBM tumors. PMID- 11380242 TI - Adenovirus-mediated transfer of p53 augments hyperthermia-induced apoptosis in U251 glioma cells. AB - PURPOSE: Hyperthermia kills glioma cells by inducing apoptosis and is thereby an effective therapeutic modality for the treatment of malignant gliomas. However, cells harboring mutated p53 are refractory to hyperthermia-induced apoptosis. In this study, we assessed whether or not adenovirus (Adv)-mediated transduction of p53 overrides this resistant mechanism. METHODS AND MATERIALS: We transduced the p53 wild-type tumor suppressor gene into U251 glioma cells harboring mutated p53 using Adv vectors in combination with hyperthermia (43, 44.5 degrees C), and evaluated the degree of cell death and apoptosis. RESULTS: The percentage of cells that had died, as measured by trypan blue staining, among U251 cells infected with the Adv for p53 (Adv-p53) and treated with hyperthermia, was significantly higher than the percentage of cells that had died among U251 cells infected with Adv-p53 and not treated with hyperthermia, or those infected with the control Adv for dE (Adv-dE) and treated with hyperthermia. The degree of apoptosis, measured at 24 h after treatment, in hyperthermia-treated U251 cells infected with Adv-p53 (43 degrees C, 73%; 44.5 degrees C, 92%) was much higher than that infected with Adv-p53 (41%), or that infected with control Adv-dE and treated with hyperthermia (43 degrees C, 1.3%; 44.5 degrees C, 19%). Treatment with combined hyperthermia and Adv-p53 infection induced cleavage of caspase-3 in U251 cells. CONCLUSION: These results indicate that Adv-mediated transduction of p53 would render glioma cells highly sensitive to hyperthermia. PMID- 11380244 TI - Combining the advantages of step-and-shoot and dynamic delivery of intensity modulated radiotherapy by interrupted dynamic sequences. AB - PURPOSE: A hybrid between step-and-shoot and dynamic operation, called interrupted dynamic sequences, was investigated for prostate intensity-modulated radiotherapy (IMRT) delivered by a multisegment close-in technique. The new delivery mode was compared to the step-and-shoot mode concerning dose distribution. METHODS AND MATERIALS: Segments suitable for dynamic transition were selected using a system of segment classes. Transitions were only allowed between two segments of the same class, keeping intended sharp in-field dose gradients unchanged. Delivery was performed by an Elekta SLiplus (Crawley, UK) linear accelerator equipped with a dynamic multileaf collimator (MLC). Because no modeling of the dose during the transitions is made, accurate dose measurements were performed. Dose profiles were measured using a linear ion chamber array (LA48, PTW-Freiburg). The suitability of this detector for measurements in sharp dose gradients was investigated first. In addition, field flatness was examined for segments with a low monitor unit (MU) count. Uncertainties in dose output were investigated using an ionization chamber (30001, PTW-Freiburg). RESULTS: Because linear array measured penumbrae are only slightly broader (< or = 0.4 mm for MLC collimated field) than those obtained using a diamond detector, the array is a good device for profile measurements. Uncertainties related with the use of low MU beam segments are very small (< 1% for segments of minimum 3 MU), giving no contra-evidence for the step-and-shoot mode. Interrupted dynamic sequences are shown to introduce only small dosimetric differences as compared to the step-and shoot delivery. CONCLUSION: Both delivery modes, step-and-shoot and interrupted dynamic sequences, result in similar dose distributions for the forward planned prostate class solution. PMID- 11380243 TI - Induction of interleukin-6 and oncostatin M by radiation in Kaposi's sarcoma cell lines. AB - PURPOSE: To define the in vitro radiosensitivity of Kaposi's sarcoma (KS) and to explore the mechanism of its extreme clinical radiosensitivity. METHODS AND MATERIALS: The radiation survivals of the three KS cell cultures (KSY-1, KS-JD, KS6-3E) were determined by clonogenic assay and MTT assay. Supernatants from irradiated cells were collected at different time points for measurement of the interleukin 6 (IL-6), oncostatin M (OSM), and basic fibroblast growth factor (bFGF) by ELISA. Changes in the mRNA expression of these cytokines were examined by reverse transcription polymerase chain reaction and Southern blot hybridization. Fresh KS cells were preincubated with the irradiated supernatant before irradiation, and the change in survivals were assessed. RESULTS: The mean SF-2 (survival fraction after 2 Gy) for KS was 0.43. Preincubation with the irradiated supernatant reduced the SF-2 significantly from 0.43 to 0.33 (p < 0.05). Treatment with irradiated supernatant alone was not cytotoxic to the cells. Radiation induced IL-6 and OSM production by KS cells at the transcription level. A single dose of 2 Gy increased IL-6 and OSM mRNA expression of the KS Y-1 cells. This corresponded to an increase in the IL-6 and OSM levels in the culture medium. There was no significant change in the level of bFGF. Preincubation with recombinant human IL-6 or OSM sensitized KS in a dose dependent manner. CONCLUSION: The low SF-2 value for these KS cell lines correlates with the clinical radiosensitivity of KS. The high radiosensitivity may be due to radiation induction of cytokines such as IL-6 and OSM, which are radiosensitizers for KS cells. PMID- 11380245 TI - Role of beam orientation optimization in intensity-modulated radiation therapy. AB - PURPOSE: To investigate the role of beam orientation optimization in intensity modulated radiation therapy (IMRT) and to examine the potential benefits of noncoplanar intensity-modulated beams. METHODS AND MATERIALS: A beam orientation optimization algorithm was implemented. For this purpose, system variables were divided into two groups: beam position (gantry and table angles) and beam profile (beamlet weights). Simulated annealing was used for beam orientation optimization and the simultaneous iterative inverse treatment planning algorithm (SIITP) for beam intensity profile optimization. Three clinical cases were studied: a localized prostate cancer, a nasopharyngeal cancer, and a paraspinal tumor. Nine fields were used for all treatments. For each case, 3 types of treatment plan optimization were performed: (1) beam intensity profiles were optimized for 9 equiangular spaced coplanar beams; (2) orientations and intensity profiles were optimized for 9 coplanar beams; (3) orientations and intensity profiles were optimized for 9 noncoplanar beams. RESULTS: For the localized prostate case, all 3 types of optimization described above resulted in dose distributions of a similar quality. For the nasopharynx case, optimized noncoplanar beams provided a significant gain in the gross tumor volume coverage. For the paraspinal case, orientation optimization using noncoplanar beams resulted in better kidney sparing and improved gross tumor volume coverage. CONCLUSION: The sensitivity of an IMRT treatment plan with respect to the selection of beam orientations varies from site to site. For some cases, the choice of beam orientations is important even when the number of beams is as large as 9. Noncoplanar beams provide an additional degree of freedom for IMRT treatment optimization and may allow for notable improvement in the quality of some complicated plans. PMID- 11380246 TI - Comparison of traditional low-dose-rate to optimized and nonoptimized high-dose rate tandem and ovoid dosimetry. AB - PURPOSE: Few dose specification guidelines exist when attempting to perform high dose-rate (HDR) dosimetry. The purpose of this study was to model low-dose-rate (LDR) dosimetry, using parameters common in HDR dosimetry, to achieve the "pear shape" dose distribution achieved with LDR tandem and ovoid applications. METHODS AND MATERIALS: Radiographs of Fletcher-Suit LDR applicators and Nucletron "Fletcher-like" HDR applicators were taken with the applicators in an idealized geometry. Traditional Fletcher loadings of 3M Cs-137 sources and the Theratronics Planning System were used for LDR dosimetry. HDR dosimetry was performed using the Nucletron Microselectron HDR UPS V11.22 with an Ir-192 source. Dose optimization points were initially located along a line 2 cm lateral to the tandem, beginning at the tandem tip at 0.5-cm intervals, ending at the sail, and optimized to 100% of the point A dose. A single dose optimization point was also placed laterally from the center of each ovoid equal to the radius of the ovoid (ovoid surface dose). For purposes of comparison, dose was also calculated for points A and B, and a point located 1 cm superior to the tandem tip in the plane of the tandem, (point F). Four- and 6-cm tandem lengths and 2.0-, 2.5-, and 3.0 cm ovoid diameters were used for this study. Based on initial findings, dose optimization schemes were developed to best approximate LDR dosimetry. Finally, radiographs were obtained of HDR applications in two patients. These radiographs were used to compare the optimization schemes with "nonoptimized" treatment plans. RESULTS: Calculated doses for points A and B were similar for LDR, optimized HDR, and nonoptimized HDR. The optimization scheme that used tapered dose points at the tandem tip and optimized a single ovoid surface point on each ovoid to 170% of point A resulted in a good approximation of LDR dosimetry. Nonoptimized HDR resulted in higher doses at point F, the bladder, and at points lateral to the tandem tip than both the optimized plan or the LDR plan. CONCLUSION: Optimized HDR allows specification of dose to points of interest, can approximate LDR dosimetry, and appears superior to nonoptimized HDR treatment planning, at least at the tandem tip. An optimization scheme is presented that approximates LDR dosimetry. PMID- 11380247 TI - Effectiveness of couch height-based patient set-up and an off-line correction protocol in prostate cancer radiotherapy. AB - PURPOSE: To investigate set-up improvement caused by applying a couch height based patient set-up method in combination with a technologist-driven off-line correction protocol in nonimmobilized radiotherapy of prostate patients. METHODS AND MATERIALS: A three-dimensional shrinking action level correction protocol is applied in two consecutive patient cohorts with different set-up methods: the traditional "laser set-up" group (n = 43) and the "couch height set-up" group (n = 112). For all directions, left-right, ventro-dorsal, and cranio-caudal, random and systematic set-up deviations were measured. RESULTS: The couch height set-up method improves the patient positioning compared to the laser set-up method. Without application of the correction protocol, both systematic and random errors reduced to 2.2-2.4 mm (1 SD) and 1.7-2.2 mm (1 SD), respectively. By using the correction protocol, systematic errors reduced further to 1.3-1.6 mm (1 SD). One dimensional deviations were within 5 mm for >90% of the measured fractions. The required number of corrections per patient in the off-line correction protocol was reduced significantly during the course of treatment from 1.1 to 0.6 by the couch height set-up method. The treatment time was not prolonged by application of the correction protocol. CONCLUSIONS: The couch height set-up method improves the set-up significantly, especially in the ventro-dorsal direction. Combination of this set-up method with an off-line correction strategy, executed by technologists, reduces the number of set-up corrections required. PMID- 11380248 TI - Remote verification in radiotherapy using digitally reconstructed radiography (DRR) and portal images: a pilot study. AB - PURPOSE: To use digitally reconstructed radiography (DRR) and digitally compressed portal images in distant consultation using a telecommunications network, the verification performance of DRR and digitally compressed portal images on the image console was investigated. METHODS AND MATERIALS: A human thoracic phantom was scanned with computed tomography (CT). Radiotherapy was planned at 5 different anatomic locations. A digitally reconstructed radiograph was made; verification films of the phantom were then taken with 6-MV X-rays. The treatment center was intentionally dislocated. Fifty sets of DRR and portal images were seen by 7 doctors on a conventional view-box (view-box method) to judge whether the treatment center was dislocated. These image sets were digitalized by a film scanner, compressed to 1/10 Joint Photographic Experts Group (JPEG) format, and compared on an image console by the same physicians (image-console method). The verification performance of the image console method was compared with that of the view-box method by means of receiver operating characteristic (ROC) analysis. Clinically, 159 portal-image-sets were verified with the image-console method and the appropriateness of the decision was later assessed by the view-box method. RESULTS: The accuracy of the treatment verification was estimated to be 88.8% by the conventional view-box method and 88.3% by the image-console method. There was no statistically significant difference in the verification performances of the conventional method (Az = 0.86+/-0.02) and the image console method (Az = 0.84+/-0.07). Frequent digital image-processing modification was positively related to the accuracy of verification. Clinically, there were 3 (1.8%) major corrections, 31 (19.5%) minor corrections, and 123 cases with no correction. No further correction was called for by the re-evaluation using the view-box method. CONCLUSION: The verification performance of DRR and digitally compressed portal images on the image console was as accurate as the conventional method. Distant consultation using DRR and portal images through telecommunication is usable in clinical practice. PMID- 11380249 TI - Structure of the transmembrane dimer interface of glycophorin A in membrane bilayers. AB - The hydrophobic transmembrane domain of glycophorin A contains a sequence motif that mediates dimerization in membrane environments. Long-range interhelical distance measurements using magic angle spinning NMR spectroscopy provide high resolution structural constraints on the packing of the dimer interface in membrane bilayers. We show that direct packing contacts occur between glycine residues at positions 79 and 83 in the transmembrane sequence. Additional interhelical constraints between Ile76 and Gly79 and between Val80 and Gly83 restrict the rotational orientation and crossing angle of the interacting helices. These results refine our previously proposed structure of the glycophorin A dimer [Smith, S. O., and Bormann, B. J. (1995) Proc. Natl. Acad. Sci. U.S.A. 92, 488-491] which revealed that the methyl groups of Val80 and Val84 are packed against Gly79 and Gly83, respectively. PMID- 11380250 TI - Main chain and side chain dynamics of a heme protein: 15N and 2H NMR relaxation studies of R. capsulatus ferrocytochrome c2. AB - A detailed characterization of the main chain and side chain dynamics in R. capsulatus ferrocytochrome c(2) derived from (2)H NMR relaxation of methyl group resonances is presented. (15)N relaxation measurements confirm earlier results indicating that R. capsulatus ferrocytochrome c(2) exhibits minor rotational anisotropy in solution. The current study is focused on the use of deuterium relaxation in side chain methyl groups, which has been shown to provide a detailed and accurate measure of internal dynamics. Results obtained indicate that the side chains of ferrocytochrome c(2) exhibit a wide range of motional amplitudes, but are more rigid than generally found in the interior of nonprosthetic group bearing globular proteins. This unusual rigidity is ascribed to the interactions of the protein with the large heme prosthetic group. This observation has significant implications for the potential of the heme-protein interface to modulate the redox properties of the protein and also points to the need for great precision in the design and engineering of heme proteins. PMID- 11380251 TI - Calcium-dependent conformation of a heme and fingerprint peptide of the diheme cytochrome c peroxidase from Paracoccus pantotrophus. AB - The structural changes in the heme macrocycle and substituents caused by binding of Ca(2+) to the diheme cytochrome c peroxidase from Paracoccus pantotrophus were clarified by resonance Raman spectroscopy of the inactive fully oxidized form of the enzyme. The changes in the macrocycle vibrational modes are consistent with a Ca(2+)-dependent increase in the out-of-plane distortion of the low-potential heme, the proposed peroxidatic heme. Most of the increase in out-of-plane distortion occurs when the high-affinity site I is occupied, but a small further increase in distortion occurs when site II is also occupied by Ca(2+) or Mg(2+). This increase in the heme distortion explains the red shift in the Soret absorption band that occurs upon Ca(2+) binding. Changes also occur in the low frequency substituent modes of the heme, indicating that a structural change in the covalently attached fingerprint pentapeptide of the LP heme occurs upon Ca(2+) binding to site I. These structural changes may lead to loss of the sixth ligand at the peroxidatic heme in the semireduced form of the enzyme and activation. PMID- 11380252 TI - Expression of the Oct-1 transcription factor and characterization of its interactions with the Bob1 coactivator. AB - The Oct-1 transcription factor regulates a variety of tissue-specific and general housekeeping genes by recruiting specialized coactivators of transcription. It acts synergistically with the B-cell-specific coactivator Bob1 (OCA-B, OBF-1) to stimulate transcription of immunoglobulin genes. To analyze Oct-1's interactions with Bob1 and other regulatory proteins, we have overexpressed and purified different functional domains of the recombinant proteins. A version of Oct-1 that encompasses the amino-terminal activation region and the POU DNA-binding domain was extensively characterized (OctDeltaC1; comprising residues 1-445). Using an in vitro transcription assay, we demonstrate that this fragment is sufficient and necessary to stimulate transcription from an immunoglobulin promoter with Bob1. It also coactivates from the herpes simplex virus ICPO promoter element in the presence of VP16. Using a range of spectroscopic and biophysical techniques, we demonstrate that the activation domains of Oct-1 and Bob1 have little globular structure and that they do not physically interact. Thus, their functional synergy is likely to arise by the co-recruitment of common factors as part of a larger regulatory assembly. We propose a hypothesis to explain why the activation domains of these and other transcription factors of metazoans have little if any intrinsic structure. PMID- 11380253 TI - Phospholipases stimulate secretion in RBL mast cells. AB - Roles for glycerophospholipids in exocytosis have been proposed, but remain controversial. Phospholipases are stimulated following the activation of the high affinity receptor for immunoglobulin E (IgE) in mast cells. To study the biochemical sequelae that lead to degranulation, broken cell systems were employed. We demonstrate that the addition of three distinct types of exogenous phospholipases (i.e., bcPLC, scPLD, and tfPLA(2)), all of which hydrolyze phosphatidylcholine (PC), trigger degranulation in permeabilized RBL-2H3 cells, a mucosal mast cell line. Production of bioactive lipids by these phospholipases promotes release of granule contents through the plasma membrane and acts downstream of PKC, PIP(2), and Rho subfamily GTPases in regulated secretion. These exogenous phospholipase-induced degranulation pathways circumvent specific factors activated following stimulation of the IgE receptor as well as in ATP- and GTP-dependent intracellular pathways. Taken together, these results suggest that regulated secretion may be achieved in vitro in the absence of cytosolic factors via phospholipase activation and that products of PC hydrolysis can promote exocytosis in mast cells. PMID- 11380254 TI - Probing catalysis by Escherichia coli dTDP-glucose-4,6-dehydratase: identification and preliminary characterization of functional amino acid residues at the active site. AB - A model of the Escherichia coli dTDP-glucose-4,6-dehydratase (4,6-dehydratase) active site has been generated by combining amino acid sequence alignment information with the 3-dimensional structure of UDP-galactose-4-epimerase. The active site configuration is consistent with the partially refined 3-dimensional structure of 4,6-dehydratase, which lacks substrate-nucleotide but contains NAD(+) (PDB file ). From the model, two groups of active site residues were identified. The first group consists of Asp135(DEH), Glu136(DEH), Glu198(DEH), Lys199(DEH), and Tyr301(DEH). These residues are near the substrate-pyranose binding pocket in the model, they are completely conserved in 4,6-dehydratase, and they differ from the corresponding equally well-conserved residues in 4 epimerase. The second group of residues is Cys187(DEH), Asn190(DEH), and His232(DEH), which form a motif on the re face of the cofactor nicotinamide binding pocket that resembles the catalytic triad of cysteine-proteases. The importance of both groups of residues was tested by mutagenesis and steady-state kinetic analysis. In all but one case, a decrease in catalytic efficiency of approximately 2 orders of magnitude below wild-type activity was observed. Mutagenesis of each of these residues, with the exception of Cys187(DEH), which showed near-wild-type activity, clearly has important negative consequences for catalysis. The allocation of specific functions to these residues and the absolute magnitude of these effects are obscured by the complex chemistry in this multistep mechanism. Tools will be needed to characterize each chemical step individually in order to assign loss of catalytic efficiency to specific residue functions. To this end, the effects of each of these variants on the initial dehydrogenation step were evaluated using a the substrate analogue dTDP-xylose. Additional steady-state techniques were employed in an attempt to further limit the assignment of rate limitation. The results are discussed within the context of the 4,6-dehydratase active site model and chemical mechanism. PMID- 11380255 TI - Mimotopes of the nicotinic receptor binding site selected by a combinatorial peptide library. AB - Peptide libraries allow selecting new molecules, defined as mimotopes, which are able to mimic the structural and functional features of a native protein. This technology can be applied for the development of new reagents, which can interfere with the action of specific ligands on their target receptors. In the present study we used a combinatorial library approach to produce synthetic peptides mimicking the snake neurotoxin binding site of nicotinic receptors. On the basis of amino acid sequence comparison of different alpha-bungarotoxin binding receptors, we designed a 14 amino acid combinatorial synthetic peptide library with five invariant, four partially variant, and five totally variant positions. Peptides were synthesized using SPOT synthesis on cellulose membranes, and binding sequences were selected using biotinylated alpha-bungarotoxin. Each variant position was systematically identified, and all possible combinations of the best reacting amino acids in each variant position were tested. The best reactive sequences were identified, produced in soluble form, and tested in BIACORE to compare their kinetic constants. We identified several different peptides that can inhibit the binding of alpha-bungarotoxin to both muscle and neuronal nicotinic receptors. Peptide mimotopes have a toxin-binding affinity that is considerably higher than peptides reproducing native receptor sequences. PMID- 11380256 TI - Asymmetric structure of the cystic fibrosis transmembrane conductance regulator chloride channel pore suggested by mutagenesis of the twelfth transmembrane region. AB - The cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel contains 12 membrane-spanning regions which are presumed to form the transmembrane pore. Although a number of findings have suggested that the sixth transmembrane region plays a key role in forming the pore and determining its functional properties, the role of other transmembrane regions is currently not well established. Here we assess the functional importance of the twelfth transmembrane region, which occupies a homologous position in the carboxy terminal half of the CFTR molecule to that of the sixth transmembrane region in the amino terminal half. Five residues in potentially important regions of the twelfth transmembrane region were mutated individually to alanines, and the function of the mutant channels was examined using patch clamp recording following expression in mammalian cell lines. Three of the five mutations significantly weakened block of unitary Cl(-) currents by SCN(-), implying a partial disruption of anion binding within the pore. Two of these mutations also caused a large reduction in the steady-state channel mean open probability, suggesting a role for the twelfth transmembrane region in channel gating. However, in direct contrast to analogous mutations in the sixth transmembrane region, all mutants studied here had negligible effects on the anion selectivity and unitary Cl(-) conductance of the channel. The relatively minor effects of these five mutations on channel permeation properties suggests that, despite their symmetrical positions within the CFTR protein, the sixth and twelfth transmembrane regions make highly asymmetric contributions to the functional properties of the pore. PMID- 11380257 TI - High-yield expression and functional analysis of Escherichia coli glycerol-3 phosphate transporter. AB - The glycerol-3-phosphate (G3P) transporter, GlpT, from Escherichia coli mediates G3P and inorganic phosphate exchange across the bacterial inner membrane. It possesses 12 transmembrane alpha-helices and is a member of the Major Facilitator Superfamily. Here we report overexpression, purification, and characterization of GlpT. Extensive optimization applied to the DNA construct and cell culture has led to a protocol yielding approximately 1.8 mg of the transporter protein per liter of E. coli culture. After purification, this protein binds substrates in detergent solution, as measured by tryptophan fluorescence quenching, and its dissociation constants for G3P, glycerol-2-phosphate, and inorganic phosphate at neutral pH are 3.64, 0.34, and 9.18 microM, respectively. It also shows transport activity upon reconstitution into proteoliposomes. The phosphate efflux rate of the transporter in the presence of G3P is measured to be 29 micromol min(-1) mg( 1) at pH 7.0 and 37 degrees C, corresponding to 24 mol of phosphate s(-1) (mol of protein)(-1). In addition, the glycerol-3-phosphate transporter is monomeric and stable over a wide pH range and in the presence of a variety of detergents. This preparation of GlpT provides ideal material for biochemical, biophysical, and structural studies of the glycerol-3-phosphate transporter. PMID- 11380258 TI - Conversion of phospholamban into a soluble pentameric helical bundle. AB - Although membrane proteins and soluble proteins may achieve their final folded states through different pathways, it has been suggested that the packing inside a membrane protein could maintain a similar fold if the lipid-exposed surface were redesigned for solubility in an aqueous environment. To test this idea, the surface of the transmembrane domain of phospholamban (PLB), a protein that forms a stable helical homopentamer within the sarcoplasmic reticulum membrane, has been redesigned by replacing its lipid-exposed hydrophobic residues with charged and polar residues. CD spectra indicate that the full-length soluble PLB is highly alpha-helical. Small-angle X-ray scattering and multiangle laser light scattering experiments reveal that this soluble variant of PLB associates as a pentamer, preserving the oligomeric state of the natural protein. Mutations that destabilize native PLB also disrupt the pentamer. However, NMR experiments suggest that the redesigned protein exhibits molten globule-like properties, possibly because the redesign of the surface of this membrane protein may have altered some native contacts at the core of the protein or possibly because the core is not rigidly packed in wild-type PLB. Nonetheless, our success in converting the membrane protein PLB into a specific soluble helical pentamer indicates that the interior of a membrane protein contains at least some of the determinants necessary to dictate folding in an aqueous environment. The design we successfully used was based on one of the two models in the literature; the alternative design did not give stable, soluble pentamers. This suggests that surface redesign can be employed in gaining insights into the structures of membrane proteins. PMID- 11380259 TI - Increase in the stability and helical content of estrogen receptor alpha in the presence of the estrogen response element: analysis by circular dichroism spectroscopy. AB - Ligand-dependent stabilization of the estrogen receptor (ER) is often postulated, with limited support from experimental data. We studied the thermal unfolding of recombinant ERalpha by circular dichroism (CD) spectroscopy. The T(M) of unfolding of ERalpha was 38 +/- 2.4 degrees C, and the van't Hoff enthalpy of unfolding was 31.7 +/- 3.4 kcal/mol in the absence of ligands. Addition of estradiol (E(2)) increased the T(M) to 43.6 +/- 2.3 degrees C, while addition of E(2) and an oligonucleotide harboring the estrogen response element (ERE) increased the T(M) to 47.9 +/- 1.6 degrees C. Addition of the antiestrogen 4 hydroxytamoxifen (HT) alone did not increase the T(M); however, a combination of HT and the ERE increased the T(M) to 48.9 +/- 1.0 degrees C. The ERE alone increased the T(M) to 46.1 +/- 0.9 degrees C. Addition of E(2) alone had no effect on the apparent enthalpy of unfolding; however, the ERE alone increased the apparent enthalpy from 31.7 to 36.1 kcal/mol. ERalpha samples containing the ERE also exhibited an increase in the negative ellipticity at 208 and 222 nm, relative to that of ligand-free ERalpha, suggesting a stabilization of the alpha helix. CD data analysis further showed that the presence of the ERE caused a large increase in alpha-helical content of ERalpha in both the presence and absence of the ligands. This increase in alpha-helical content of ERalpha was not observed in the presence of a nonspecific oligonucleotide. These results show that the ERE can increase the thermal stability of ERalpha, enhance its alpha helical content, and facilitate the cooperativity of the folding transition. PMID- 11380260 TI - Human endonuclease III acts preferentially on DNA damage opposite guanine residues in DNA. AB - The human endonuclease III homologue (hNTH1) removes premutagenic cytosine damage from DNA. This includes 5-hydroxycytosine, which has increased potential for pairing with adenine, resulting in C --> T transition mutations. Here we report that hNTH1 acts on both 5-hydroxycytosine and abasic sites preferentially when these are situated opposite guanines in DNA. Discrimination against other opposite bases is strongly dependent on the presence of magnesium. To further elucidate this effect, we have introduced mutations in the helix-hairpin-helix domain of hNTH1 (K212S, P211R, +G212, and DeltaP211), and measured the kinetics of 5-hydroxycytosine removal of the mutants relative to wild type. The K212S and DeltaP211 (truncated hairpin) mutant proteins were both inactive, whereas the extended hairpin in the +G212 mutant diminished recognition and binding to 5 hydroxycytosine-containing DNA. The P211R mutant resembled native hNTH1, except for decreased specificity of binding. Despite the altered kinetic parameters, the active mutants retained the ability to discriminate against the pairing base, indicating that enzyme interactions with the opposite strand relies on other domains than the active site helix-hairpin-helix motif. PMID- 11380261 TI - Base sequence dependence of in vitro translesional DNA replication past a bulky lesion catalyzed by the exo- Klenow fragment of Pol I. AB - The effects of base sequence, specifically different pyrimidines flanking a bulky DNA adduct, on translesional synthesis in vitro catalyzed by the Klenow fragment of Escherichia coli Pol I (exo(-)) was investigated. The bulky lesion was derived from the binding of a benzo[a]pyrene diol epoxide isomer [(+)-anti-BPDE] to N(2) guanine (G*). Four different 43-base long oligonucleotide templates were constructed with G* at a site 19 bases from the 5'-end. All bases were identical, except for the pyrimidines, X or Y, flanking G* (sequence context 5'-.XGY., with X, Y = C and/or T). In all cases, the adduct G* slows primer extension beyond G* more than it slows the insertion of a dNTP opposite G* (A and G were predominantly inserted opposite G, with A > G). Depending on X or Y, full lesion bypass differed by factors of approximately 1.5-5 ( approximately 0.6-3.0% bypass efficiencies). A downstream T flanking G on the 5'-side instead of C favors full lesion bypass, while an upstream C flanking G* is more favorable than a T. Various deletion products resulting from misaligned template-primer intermediates are particularly dominant ( approximately 5.0-6.0% efficiencies) with an upstream flanking C, while a 3'-flanking T lowers the levels of deletion products ( approximately 0.5-2.5% efficiencies). The kinetics of (1) single dNTP insertion opposite G* and (2) extension of the primer beyond G* by a single dNTP, or in the presence of all four dNTPs, with different 3'-terminal primer bases (Z) opposite G* were investigated. Unusually efficient primer extension efficiencies beyond the adduct (approaching approximately 90%) was found with Z = T in the case of sequences with 3'-flanking upstream C rather than T. These effects are traced to misaligned slipped frameshift intermediates arising from the pairing of pairs of downstream template base sequences (up to 4-6 bases from G*) with the 3'-terminal primer base and its 5'-flanking base. The latter depend on the base Y and on the base preferentially inserted opposite the adduct. Thus, downstream template sequences as well as the bases flanking G* influence DNA translesion synthesis. PMID- 11380262 TI - The antithrombin P1 residue is important for target proteinase specificity but not for heparin activation of the serpin. Characterization of P1 antithrombin variants with altered proteinase specificity but normal heparin activation. AB - Heparin has been proposed to conformationally activate the serpin, antithrombin, by making the reactive center loop P1 arginine residue accessible to proteinases. To evaluate this proposal, we determined the effect of mutating the P1 arginine on antithrombin's specificity for target and nontarget proteinases in both native and heparin-activated states of the serpin. As expected, mutation of the P1 arginine to tryptophan, histidine, leucine, and methionine converted the specificity of antithrombin from a trypsin inhibitor (k(assoc) = 2 x 10(5) M(-1) s(-1)) to a chymotrypsin inhibitor (k(assoc) = 10(3)-10(5) M(-1) s(-1)). However, heparin pentasaccharide activation increased the reactivity of the P1 variants with chymotrypsin or of the wild-type inhibitor with trypsin only 2-6-fold, implying that the P1 residue had similar accessibilities to these proteinases in native and activated states. Mutation of the P1 arginine greatly reduced k(assoc) for antithrombin inhibition of thrombin and factor Xa from 40- to 5000-fold, but heparin normally accelerated the reactions of the variant antithrombins with these enzymes to make them reasonably efficient inhibitors (k(assoc) = 10(3) 10(4) M(-1) s(-1)). Fluorescence difference spectra of wild-type and P1 tryptophan variant antithrombins showed that the P1 tryptophan exhibited fluorescence properties characteristic of a solvent-exposed residue which were insignificantly affected by heparin activation. Moreover, all P1 variant antithrombins bound heparin with approximately 2-3-fold higher affinities than the wild type. These findings are consistent with the P1 mutations disrupting a P1 arginine-serpin body interaction which stabilizes the native low-heparin affinity conformation, but suggest that this interaction is of low energy and unlikely to limit the accessibility of the P1 residue. Together, these findings suggest that the P1 arginine residue is similarly accessible to proteinases in both native and heparin-activated states of the serpin and contributes similarly to the specificity of antithrombin for thrombin and factor Xa in the two serpin conformational states. Consequently, determinants other than the P1 residue are responsible for enhancing the specificity of antithrombin for the two proteinases when activated by heparin. PMID- 11380263 TI - Conformational equilibrium of the reactive center loop of antithrombin examined by steady state and time-resolved fluorescence measurements: consequences for the mechanism of factor Xa inhibition by antithrombin-heparin complexes. AB - Activation of antithrombin by high-affinity heparin as an inhibitor of factor Xa has been ascribed to an allosteric switch between two conformations of the reactive center loop. However, we have previously shown that other, weaker binding, charged polysaccharides can give intermediate degrees of activation [Gettins, P. G. W., et al. (1993) Biochemistry 32, 8385-8389]. To examine whether such intermediate activation results from different reactive center loop conformations or, more simply, from a different equilibrium constant between the same two extreme conformations, we have used NBD covalently bound at the P1 position of an engineered R393C variant of antithrombin as a fluorescent reporter group and measured fluorescence lifetimes of the label in free antithrombin as well as in antithrombin saturated with long-chain high-affinity heparin, high affinity heparin pentasaccharide, long-chain low-affinity heparin, and dextran sulfate. Steady state emission spectra, anisotropies, and dynamic quenching measurements were also recorded. We found that the large steady state fluorescence enhancements produced by binding of activators resulted from relief of a static quench of fluorescence of NBD in approximately 50% of the labeled antithrombin molecules rather than from any large change in lifetimes, and that similar lifetimes were found for NBD in all activated antithrombin oligosaccharide complexes. Similar anisotropies and positions of the NBD emission maxima were also found in the absence and presence of activators. In addition, NBD was accessible to quenching agents in both the absence and presence of activators, with an at most 2-fold increase in quenching constants between these two extremes. The simplest interpretation of the partial static quench in the absence of activators, the different degrees of enhancement by different antithrombin activators, and the similar fluorescence properties and quenching behavior of the different states is that there are two distinct types of conformational equilibrium involving three distinct states of antithrombin, which we designate A, A', and B. A and A' represent low-affinity or inactive states of approximately equal energy, both having the hinge residues inserted into beta sheet A. A is fluorescent, while A' is statically quenched. State B represents the activated loop-expelled conformation in which none of the NBD fluorophores are statically quenched, as a result of the loop, including the P1-NBD, moving away from the body of the antithrombin. Different activators are able to shift the equilibrium to the high-activity (B) state to different extents and hence give different degrees of measured activity, and different degrees of relief of static quench. The similar properties and accessibility of the NBD in the A and B conformations also indicate that the P1 side chain is not buried in the low activity A conformation, suggesting that an earlier proposal that activation involves exposure of the P1 side chain cannot be the explanation for activation. As an alternative explanation, heparin activation may give access to an exosite on antithrombin for binding to factor Xa and hence be the principal basis for enhancement of the rate of inhibition. PMID- 11380264 TI - Mg2+-induced tRNA folding. AB - Mg(2+)-induced folding of yeast tRNA(Phe) was examined at low ionic strength in steady-state and kinetic experiments. By using fluorescent labels attached to tRNA, four conformational transitions were revealed when the Mg(2+) concentration was gradually increased. The last two transitions were not accompanied by changes in the number of base pairs. The observed transitions were attributed to Mg(2+) binding to four distinct types of sites. The first two types are strong sites with K(diss) of 4 and 16 microM. The sites of the third and fourth types are weak with a K(diss) of 2 and 20 mM. Accordingly, the Mg(2+)-binding sites previously classified as "strong" and "weak" can be further subdivided into two subtypes each. Fluorescent transition I is likely to correspond to Mg(2+) binding to a unique strong site selective for Mg(2+); binding to this site causes only minor A(260) change. The transition at 2 mM Mg(2+) is accompanied by substantial conformational changes revealed by probing with ribonucleases T1 and V1 and likely enhances stacking of the tRNA bases. Fast and slow kinetic phases of tRNA refolding were observed. Time-resolved monitoring of Mg(2+) binding to tRNA suggested that the slow kinetic phase was caused by a misfolded tRNA structure formed in the absence of Mg(2+). Our results suggest that, similarly to large RNAs, Mg(2+)-induced tRNA folding exhibits parallel folding pathways and the existence of kinetically trapped intermediates stabilized by Mg(2+). A multistep scheme for Mg(2+)-induced tRNA folding is discussed. PMID- 11380265 TI - Acid-base catalysis by UDP-galactose 4-epimerase: correlations of kinetically measured acid dissociation constants with thermodynamic values for tyrosine 149. AB - The steady-state kinetic parameters for epimerization of UDP-galactose by UDP galactose 4-epimerase from Escherichia coli (GalE), Y149F-GalE, and S124A-GalE have been measured as a function of pH. The deuterium kinetic isotope effects for epimerization of UDP-galactose-C-d(7) by these enzymes have also been measured. The results show that the activity of wild-type GalE is pH-independent in the pH range of 5.5-9.3, and there is no significant deuterium kinetic isotope effect in the reaction of UDP-galactose-C-d(7). It is concluded that the rate-limiting step for epimerization by wild-type GalE is not hydride transfer and must be either a diffusional process or a conformational change. Epimerization of UDP-galactose-C d(7) by Y149F-GalE proceeds with a pH-dependent deuterium kinetic isotope effect on k(cat) of 2.2 +/- 0.4 at pH 6.2 and 1.1 +/- 0.5 at pH 8.3. Moreover, the plot of log k(cat)/K(m) breaks downward on the acid side with a fitted value of 7.1 for the pK(a). It is concluded that the break in the pH-rate profile arises from a change in the rate-limiting step from hydride transfer at low pH to a conformational change at high pH. Epimerization of UDP-galactose-C-d(7) by S124A GalE proceeds with a pH-independent deuterium kinetic isotope effect on k(cat) of 2.0 +/- 0.2 between pH 6 and 9. Both plots of log k(cat) and log k(cat)/K(m) display pH dependence. The plot of log k(cat) versus pH breaks downward with a pK(a) of 6.35 +/- 0.10. The plot of log k(cat)/K(m) versus pH is bell-shaped, with fitted pK(a) values of 6.76 +/- 0.09 and 9.32 +/- 0.21. It is concluded that hydride transfer is rate-limiting, and the pK(a) of 6.7 for free S124A-GalE is assigned to Tyr 149, which displays the same value of pK(a) when measured spectrophotometrically in this variant. Acid-base catalysis by Y149F-GalE is attributed to Ser 124, which is postulated to rescue catalysis of proton transfer in the absence of Tyr 149. The kinetic pK(a) of 7.1 for free Y149F-GalE is lower than that expected for Ser 124, as proven by the pH-dependent kinetic isotope effect. Epimerization by the doubly mutated Y149F/S124A-GalE proceeds at a k(cat) that is lower by a factor of 10(7) than that of wild-type GalE. This low rate is attributed to the synergistic actions of Tyr 149 and Ser 124 in wild-type GalE and to the absence of any internal catalysis of hydride transfer in the doubly mutated enzyme. PMID- 11380267 TI - An historical perspective on genetic care. AB - The outcomes of genetic research endeavors have the potential to transform health care with significant implications for both providers and consumers of clinical services. Professional and public integration of genetics knowledge is key to successful utilization of genetics information. This article will provide an overview of genetics including a historical perspective, examples of genetic health care, the nursing perspective, and ethical considerations and challenges. New scientific explanations for health, disease, responsiveness to treatment, and design of options for care may create personal and professional dilemmas. Nurses have a responsibility to become active participants in confronting the demands resulting from this new knowledge for education, practice, and policy. The purpose of this article is to provide a foundation from which the profession of nursing can build to enhance current skills and knowledge about genetics to prepare for this transformation in health care. PMID- 11380268 TI - Prenatal genetic screening. AB - This article presents a discussion of screening principles and techniques available to screen for common birth defects during pregnancy. Sixty-five to 70% of women have serum screening and /or ultrasound during pregnancy to evaluate the health and well-being of the developing fetus. The most common birth defects identified by screening include neural tube defects and chromosome abnormalities. Nurses employed in prenatal care settings need to have accurate information they can provide to women so they understand the benefits and limitations of screening. Timely presentation of information and identification of available resources will help nurses minimize confusion and provide support for women as they proceed with pregnancy screening. PMID- 11380269 TI - Genetic testing and mental health: the model of Huntington disease. AB - Genetic aspects of mental health disorders are being identified through human genome and family research. Gene discovery makes diagnostic and presymptomatic testing possible. The discovery of a gene mutation for Huntington Disease (HD) enables at-risk persons to request presymptomatic genetic testing. When HD genetic testing is offered through HD testing centers, a multi-visit protocol is followed in which education and counseling are provided for persons considering the option to have HD gene testing. A case study illustrates the clinical and ethical issues regarding privacy and disclosure as well as the personal and family consequences of gene mutation knowledge. Analysis of the impact of genetic knowledge on persons being tested for HD provides a model for the integration of emerging genetic information into mental health nursing practice for other mental health disorders. PMID- 11380270 TI - Heart stopper genes: would you recognize a high risk patient? AB - Coronary artery disease (CAD) is one of the leading causes of death in the United States. Eighty percent of people having heart attacks have normal cholesterol levels. A quarter of the population have a condition called low-density lipoprotein (LDL) pattern B that has been associated with a threefold risk of myocardial infarction. Although early intervention can often prevent an otherwise fatal event, these patients often go unrecognized until after a myocardial infarction has occurred because they may not have the usual risk factors associated with cardiovascular disease. In patients with LDL pattern B, the standard lipid panel may be normal and inadequate for diagnosis and treatment of the condition. This article discusses how to identify a potentially high risk patient, available laboratory tests, management options, and the role of nurses in identifying high risk patients. The second author tells his personal story of surviving multiple cardiac arrests at a young age before being diagnosed with this condition. PMID- 11380271 TI - Ethical implications of genetic information. PMID- 11380272 TI - A new world view of genetics service models. AB - Identification of the components of the Human Genome and their relevance to health and disease is revolutionizing the provision of genetic services and all areas of health care. New genetic tools to diagnose, manage, and treat common diseases, along with Web-based innovations are changing the shape of how genetic services will be accessed and delivered. These advances represent a "bionic convergence" that will fundamentally transform medicine during the next few decades. The convergence of biotechnology and electronics is creating an expanding array of health care opportunities for clients and offers innovative opportunities for health promotion, restoration and management. Families and communities will soon be able to participate more fully in the direction and design of their own genetic health. Nurses with their long history of providing holistic, family-centered care in all practice settings, can help to create new dimensions to their practice to support their clients as they meet these health care innovations. This article explores a New World view of genetics services, and describes futuristic models for their provision. Nursing participation in and preparation for future genetics services also is described. PMID- 11380273 TI - Challenging myths in physical therapy. PMID- 11380274 TI - Effect of burst-mode transcutaneous electrical nerve stimulation on peripheral vascular resistance. AB - BACKGROUND AND PURPOSE: Based on changes in skin temperature alone, some authors have proposed that postganglionic sympathetic vasoconstrictor fibers can be stimulated transcutaneously. Our goal was to determine the effects of low frequency (2 bursts per second), burst-mode transcutaneous electrical nerve stimulation (TENS) on calf vascular resistance, a more direct marker of sympathetic vasoconstrictor outflow than skin temperature, in subjects with no known pathology. SUBJECTS: Fourteen women and 6 men (mean age=31 years, SD=13, range=18-58) participated in this study. METHODS: Calf blood flow, arterial pressure, and skin temperature were measured while TENS was applied over the common peroneal and tibial nerves. RESULTS: Blood flow immediately following stimulation was not affected by TENS applied just under or just above the threshold for muscle contraction. Transcutaneous electrical nerve stimulation applied at 25% above the motor threshold caused a transient increase in calf blood flow. Regardless of stimulation intensity, TENS had no effect on arterial pressure; therefore, calf vascular resistance decreased only during the trial that was 25% above the motor threshold. Regardless of stimulation intensity, TENS failed to alter dorsal or plantar skin temperature. DISCUSSION AND CONCLUSION: These results demonstrate that the effects of TENS on circulation depend on stimulation intensity. When the intensity was sufficient to cause a moderate muscle contraction, a transient, local increase in blood flow occurred. Cooling of the dorsal and plantar skin occurred in both the stimulated and control legs, most likely because skin temperature acclimatized to ambient room temperature, rather than because of any effect of TENS on circulation. The data, therefore, call into question the idea that postganglionic sympathetic efferent fibers are stimulated when TENS is applied at clinically relevant intensities to people without symptoms of cardiovascular or neuromuscular pathology. PMID- 11380275 TI - Development and validation of a telephone questionnaire to characterize lymphedema in women treated for breast cancer. AB - BACKGROUND AND PURPOSE: Accurate and economical characterization of lymphedema is needed for population-based studies of incidence and risk. The purpose of this study was to develop and validate a telephone questionnaire for characterizing lymphedema. SUBJECTS: Forty-three women who were treated previously for breast cancer and who were recruited from physical therapy practices and a cancer support organization were studied. METHODS: Questionnaire assessment of the presence and degree of lymphedema was compared with physical therapists' diagnoses, based primarily on circumferential measurements. Twenty-five of the 43 subjects were measured independently by 2 physical therapists to assess interobserver agreement. RESULTS: Interobserver agreement on clinical assessments of the presence and degree of lymphedema was high (20/25, weighted kappa=.80); all of the disagreements were between judgments of whether there was no lymphedema or mild lymphedema. For the diagnosis of at least moderate lymphedema (differential in the circumferences of the upper extremities greater than 2 cm), sensitivity of the questionnaire varied from 0.86 to 0.92 and specificity was 0.90. However, sensitivity (varying from 0.93 to 0.96) was higher than specificity (varying from 0.69 to 0.75) for the diagnosis of any lymphedema. DISCUSSION AND CONCLUSION: A few straightforward questions exhibited excellent agreement with physical therapists' assessments for identifying at least moderate lymphedema. PMID- 11380276 TI - Effect of superficial heat, deep heat, and active exercise warm-up on the extensibility of the plantar flexors. AB - BACKGROUND AND PURPOSE: Warm-up prior to static stretching enhances muscle extensibility. The relative effectiveness of different modes of warm-up, however, is unknown. The purpose of this study was to evaluate the effectiveness of superficial heat, deep heat, and active exercise warm-up prior to stretching compared with stretching alone on the extensibility of the plantar-flexor muscles. SUBJECTS: Ninety-seven subjects (59 women, 38 men) with limited dorsiflexion range of motion (ROM) were randomly assigned to 1 of 5 groups. Female subjects had a mean age of 27.6 years (SD=7.68, range=17-50), and male subjects had a mean age of 26.8 years (SD=6.87, range=18-48). METHODS: The first group (group 1) was a control group and did not perform the stretching protocol. The 4 experimental groups (groups 2-5) performed a stretching protocol 3 days per week for 6 weeks. Group 2 performed the static stretching protocol only; group 3 performed active heel raises before stretching; group 4 received 15 minutes of superficial, moist heat to the plantar-flexor muscles before stretching; and group 5 received continuous ultrasound for 7 minutes before stretching. Dorsiflexion ROM measurements were taken initially and after 2, 4, and 6 weeks. RESULTS: All experimental groups increased active and passive range of motion (AROM and PROM). The mean AROM/PROM differences at 6 weeks were 1.11/1.39 degrees for group 1, 4.10/6.11 degrees for group 2, 4.16/4.21 degrees for group 3, 4.38/4.90 degrees for group 4, and 6.20/7.35 degrees for group 5. The group receiving ultrasound before performing the stretching protocol (group 5) displayed the greatest increase in both AROM (6.20() and PROM (7.35(). DISCUSSION AND CONCLUSION: Among the modalities tested, the use of ultrasound for 7 minutes prior to stretching may be the most effective for increasing ankle dorsiflexion ROM. PMID- 11380277 TI - The effect of quadriceps femoris muscle strengthening exercises on spasticity in children with cerebral palsy. AB - BACKGROUND AND PURPOSE: The Bobath neurodevelopmental treatment approach advised against the use of resistive exercise, as proponents felt that increased effort would increase spasticity. The purpose of this study was to test the premise that the performance of exercises with maximum efforts will increase spasticity in people with cerebral palsy (CP). Spasticity, in the present study, was defined as a velocity-dependent hyperexcitability of the muscle stretch reflex. SUBJECTS: Twenty-four subjects with the spastic diplegic form of CP (mean age=11.4 years, SD=3.0, range=7-17) and 12 subjects without known neurological impairments (mean age=11.6 years, SD=3.5, range=7-17) were assessed. METHODS: Knee muscle spasticity was assessed bilaterally using the pendulum test to elicit a stretch reflex immediately before and after 3 different forms of right quadriceps femoris muscle exercise (isometric, isotonic, and isokinetic) during a single bout of exercise training. Pendulum test outcome measures were: (1) first swing excursion, (2) number of lower leg oscillations, and (3) duration of the oscillations. RESULTS: There were no changes in spasticity following exercise between the 2 groups of subjects. DISCUSSION AND CONCLUSION: These results do not support the premise that exercises with maximum efforts increase spasticity in people with CP. PMID- 11380278 TI - Reasons given by California physical therapists for not belonging to the american physical therapy association. AB - BACKGROUND AND PURPOSE: In 1997, only 22% of licensed physical therapists living in California were members of the American Physical Therapy Association (APTA). This 1998 study was designed to identify the reason(s) why most licensed physical therapists in California choose not to belong to their profession's national association and to examine the demographics of nonmembers. SUBJECTS: The subjects were a random sample of 400 California licensed physical therapists who were not members of APTA. METHODS: The survey instrument included a demographic questionnaire and statements for response using a 5-point Likert-type scale. Frequency distributions were calculated for responses and demographic data. Nonparametric analyses were used to determine statistical significance. Chi square analysis was used to compare responses to statements by gender and by full time versus part-time work status. Spearman rank correlation coefficients were used to determine any relationships between demographic data (eg, gender and work status). The Mann-Whitney U test was used to determine any differences in responses to specific representation questions by those respondents who worked in those environments. All statistical tests were 2-tailed tests conducted at the P(.05 level, unless otherwise indicated. Means, standard deviations, and ranges were used where appropriate. RESULTS: There was a 67% response rate. Eighty-nine percent of the respondents had been members of APTA. Eighty-eight percent of the respondents believed that APTA national dues were too high, and 90% thought California Chapter dues were too high. DISCUSSION AND CONCLUSION: Cost was the primary reason given for APTA nonmembership in California. PMID- 11380279 TI - Making geriatric assessment work: selecting useful measures. AB - Often the goal of physical therapy is to reduce morbidity and prevent or delay loss of independence. The purpose of this article is to describe issues to consider when selecting measures of physical function for use with community dwelling older adults over the age of 65 years. We chose 16 measures of physical function for review because they have been used in studies of community-dwelling older adults and some psychometric properties of reliability and validity have been described in the literature. Three major issues are discussed: (1) appropriateness of the measure for community-dwelling adults, (2) practical aspects of test administration, and (3) psychometric properties. These issues are illustrated using examples from the 16 measures. Two scenarios, applying the measures to the assessment of community-dwelling well older people and to the assessment of community-dwelling frail older people, are used to illustrate how this information can be used. PMID- 11380280 TI - Physical therapist management of tuberculous arthritis of the elbow. AB - BACKGROUND AND PURPOSE: Tuberculous arthritis is not commonly seen by physical therapists in the United States. The purpose of this case report is to describe a case of tuberculous arthritis of the elbow. CASE DESCRIPTION: The patient was a 36-year-old man referred for physical therapy evaluation and intervention for chronic elbow pain. After an evaluation and a trial of physical therapy, the patient was referred back to a primary care provider for additional tests to rule out systemic pathology. An open debridement of synovium and biopsy of the capitellum and radial head was positive for acid-fast bacilli, which was later identified as Mycobacterium tuberculosis. OUTCOMES: The patient was placed on a 4 drug antituberculosis regimen that resolved all patient complaints and restored full elbow function. DISCUSSION: Tuberculous arthritis has characteristic findings during examination and in diagnostic tests. Although tuberculous arthritis is uncommon, it should be considered when patients have chronic or vague musculoskeletal complaints. PMID- 11380281 TI - Blood supply to the spinal cord. PMID- 11380282 TI - An introduction to gene therapy and its potential prenatal use. AB - This review article is an introduction to the general field of gene therapy with examples from the specialized field of prenatal gene therapy. PMID- 11380283 TI - Changes in molecular weight of hyaluronan and hyaluronidase activity in uterine cervical mucus in cervical ripening. AB - BACKGROUND: This study was undertaken to investigate changes in the forms of hyaluronan and hyaluronidase activity in cervical mucus during cervical ripening. METHODS: Uterine cervical mucus was obtained from 57 pregnant women (25 at preterm gestation, ten at term gestation, 11 within 1 week before labor, and 11 during the first stage of labor). We determined 1) concentration of hyaluronan, 2) hyaluronidase activity, and 3) molecular weight of hyaluronan in cervical mucus. Data are presented as mean and range. RESULTS: The hyaluronan concentration in mucus in the 1st stage of labor (1.58 microg/ml, 0.46-23.96) was significantly (p<0.05) higher than that in all other groups (preterm: 0.29, 0.10 0.88; term: 0.24, 0.11-0.80; within 1 week before labor: 0.30, 0.18-0.62). Hyaluronidase activity both within 1 week before labor group (3.03 min., 1.12 3.95) and in 1st stage of labor group (3.52, 0.43-5.15) was significantly (p<0.05) higher than that in preterm group (1.70, 0.00-5.47). The molecular weight of hyaluronan in cervical mucus in the 1st stage of labor (0.97x106, 0.86 1.41) was significantly (p<0.05) lower than in the preterm and term groups (preterm: 1.60, 1.21-2.20, term: 1.41, 1.21-2.20). There was a significant correlation between hyaluronidase activity and molecular weight of hyaluronan (p<0.05, r=-0.41, n=23). CONCLUSION: These findings suggest that either hyaluronidase or low-molecular weight hyaluronan could be one of the most important regulators in the process of cervical ripening. PMID- 11380284 TI - Expression of basic fibroblast growth factor (bFGF), FGF receptor 1 and FGF receptor 2 in uterine leiomyomas and myometrium during the menstrual cycle, after menopause and GnRHa treatment. AB - BACKGROUND: To investigate whether basic fibroblast growth factor (bFGF) is involved in the growth regulation of human uterine leiomyomas the expression of bFGF and its receptors was measured in leiomyomas and myometrium obtained under different endocrine conditions. METHODS: The expression of bFGF, fibroblast growth factor receptor 1 (FGFR1) and fibroblast growth factor receptor 2 (FGFR2) was analyzed by immunohistochemistry and Western blot. RESULTS: Twenty-seven women with leiomyomas included eight in the proliferative phase, seven in the secretory phase, six after menopause and six after GnRHa treatment. In the proliferative phase, bFGF staining in leiomyomas was significantly stronger than in any other leiomyoma group. After GnRHa treatment, the expression of bFGF in both leiomyomas and myometrium was weaker than in the proliferative phase. The staining of FGFR1 was less intense in proliferative phase myometrium than in myometrium from any other group, significantly weaker than in the secretory phase. The leiomyomas demonstrated homogeneous cytoplasmic FGFR1 staining that was similar in all groups, except in the GnRHa treated patients where a more intense staining was observed, significantly stronger than in proliferative phase leiomyomas. No tissue differences were observed for staining of FGFR2 and no significant differences were observed between the different groups. Slightly less staining of FGFR2 was found in leiomyomas in the secretory phase but it did not reach statistical significance. The specificity of immunostaining was confirmed by Western blot. CONCLUSIONS: We suggest that the regulation of bFGF, and to some extent also its receptors in leiomyomas and in myometrium, is influenced by sex steroid hormones. However, the lack of differences in expression between leiomyomas and myometrium favors the view that bFGF does not necessarily contribute to the differences in growth regulation in these tissues. PMID- 11380285 TI - Prognosis in four syndromes of pregnancy-related pelvic pain. AB - BACKGROUND: The aim of the present study was to describe, on the basis of specific classification criteria and for a period of two years after delivery, the prognosis for women suffering from pregnancy-related pelvic joint pain, and to describe the characteristics influencing the prognosis. METHODS: One thousand seven hundred and eighty-nine pregnant women who were booked for delivery at Odense University Hospital formed a cohort to investigate the prognosis. Women whose reported daily pain from pelvic joints could be objectively confirmed were divided, according to symptoms, into five subgroups (n=405) - four classification groups (pelvic girdle syndrome, symphysiolysis, one-sided sacroiliac syndrome and double-sided sacroiliac syndrome) and one miscellaneous. The women in the five subgroups were re-examined at regular intervals for two years after delivery or until disappearance of symptoms (whichever was less). Thre hundred and forty-one women from the 5 subgroups participated in the postpartum follow-up. RESULTS: The majority (62.5%) of women in the four classification groups experienced disappearance of pain within a month after delivery. Two years after parturition 8.6% were still suffering from pelvic joint pain (determined subjectively and objectively). Persistence of pain was found to vary significantly from one classification group to another. None of those initially classified as suffering from symphysiolysis had pain 6 months after delivery in comparison to the 21 percent of those with pelvic girdle syndrome who continued to have pain at the two-year mark. CONCLUSIONS: This study shows that pregnancy-related pelvic joint pain had an excellent postpartum prognosis (in general) in three out of four classification groups. The women with pelvic girdle syndrome (pain in all 3 pelvic joints) had a markedly worse prognosis than the women in the other three classification groups. High number of positive test and a low mobility index were identified as giving the highest relative risk for long term pain. PMID- 11380286 TI - Intrauterine infection may be a major cause of stillbirth in Sweden. AB - AIM OF THE STUDY: To investigate intrauterine infection as a cause for unexplained stillbirth. METHODS: Chorioamnionitis was studied in a material of stillbirths (117 subjects from the years 1985-1994) from a region in the south Sweden. Control material (126 alive and healthy newborns and with healthy mothers) was gathered from the same region. RESULTS: Chorioamnionitis was a common diagnosis both with stillbirths and 'healthy' deliveries (82 and 68%, respectively). Extension of the inflammation to decidua basalis was seven times more common among stillbirths than among controls (odds ratio 7.2, confidence interval 2.8-21.9). The most common bacteria found at cultures were Escherichia coli, Coagulase negative staphylococcus, Enterococcus faecalis and group B Streptococcus. The risk for stillbirth was doubled if both inflammation and bacteria were present (odds ratio 2.3, confidence interval 0.92-5.8). Meconium discharge was more common among stillbirths than controls (odds ratio=4.7, confidence interval 1.7-14). There were no differences in any respect regarding macerated and non-macerated stillbirths. Our findings are similar to the results from studies in developing countries except for the higher incidence of stillbirths in such countries. CONCLUSIONS: Thus, a large part of otherwise unexplained stillbirths might be due to ascending infections. PMID- 11380287 TI - Are thyroid hormones or hCG responsible for hyperemesis gravidarum? A matched paired study in pregnant Chinese women. AB - OBJECTIVE: To determine the relative importance of thyroid hormones and human chorionic gonadotropin in the etiology of hyperemesis gravidarum. DESIGN: A prospective study comparing the hormonal status in women with hyperemesis gravidarum (5-18 weeks), with healthy pregnant controls, matched for gestational age. Sensitive thyroid stimulating hormone, free thyroxine, free triiodothyronine and total beta human chorionic gonadotropin were measured by immunoassays. The hormone results for hyperemesis gravidarum group (n=58) were compared with pregnant control women (n=58) using the Mann Whitney test. Stepwise logistic regression analysis was performed to determine which variables were significantly associated with hyperemesis gravidarum and to estimate the probability of each woman having hyperemesis gravidarum. The strength of the resulting association was tested by generating a receiver operating characteristic curve for predicting hyperemesis gravidarum using these probabilities. RESULTS: Maternal age and all hormones were significantly different between the hyperemetic and control groups. However, logistic regression analysis demonstrated that only maternal age, free thyroxine and thyroid stimulating hormone were significant independent variables. The area beneath the receiver operating characteristic curve for prediction of hyperemesis gravidarum was 0.84. CONCLUSION: Human chorionic gonadotropin is not independently involved in the etiology of hyperemesis gravidarum but may be indirectly involved by its ability to stimulate the thyroid. Differences in maternal age and thyroid function are highly discriminatory with regard to hyperemesis gravidarum. PMID- 11380288 TI - The very preterm infant - a population-based study. AB - OBJECTIVES: The aim of this study was to assess perinatal risk factors and the survival of the very preterm infant in comparison with birth beyond 32nd birthweek, as well as health care utilization by mothers and infants in the Northern Health Region of Sweden. DESIGN: A population-based study was designed of all children (66,646) born in the Northern Health Region of Sweden during 1991 1996 and registered in the Swedish Medical Birth Registry. METHODS: Maternal and perinatal factors of infants born very preterm, that is, at < or =27 and 28-31 weeks of gestation, were analyzed for relative risk (RR), and a 95% confidence interval (CI), and compared with those of infants born 32-36 weeks of gestation. RESULTS: Of the 66,646 infants registered, 3,493 (5.2%) were born at 32-36 weeks, 394 (0.6%) at 28-31 weeks, and 199 (0.3%) at 22-27 weeks' gestation. No special socio-demographic maternal factors characterized these preterm births. The very preterm infants were more prone to perinatal complications such as premature rupture of the membranes (PROM) (RR=4.13; 95% CI=3.07-5.55), and both PROM and hemorrhage (RR=7.80; 95% CI=3.43-17.72). Infants born very preterm were more often twins, growth-retarded, malformed, and affected by sepsis and respiratory distress. There was significantly better survival of preterm infants born at < or =27 weeks' gestation if their mothers were given tertiary perinatal care. For infants born extremely preterm, survival tended to be better if they were delivered by cesarean section. CONCLUSION: The very preterm birth is more often than not a result of a complicated pregnancy. The infant is often sick before birth, and for its survival is highly dependent on the highest level of perinatal care. PMID- 11380289 TI - Choice and chance: determinants of short interpregnancy intervals in Denmark. AB - BACKGROUND: It is still unclear whether short interpregnancy intervals are a marker for women at risk of poor pregnancy outcome or a direct risk factor for poor perinatal outcomes. The study objective was to identify risk factors associated with short interpregnancy intervals in Denmark. METHODS: From a cohort of pregnant women in a geographically defined area in Denmark (n=11,288) and using register linkage, we identified 5756 multiparous mothers who completed a detailed interview on social behavior during pregnancy. We restricted our analysis to 2904 mothers who had an interpregnancy interval of less than 37 months. Multiple logistic regression was used to estimate the Odds Ratio (OR) of having a short interval as a function of a number of determinants. RESULTS: About 4.8% of the mothers had an interpregnancy interval less than 9 months. Short interpregnancy intervals were more likely to occur in an unplanned pregnancy (OR=2.9, 95% CI: 2.2-3.9), to follow irregular menstruation (OR=1.7, 95% CI: 1.1, 2.5) and to occur in older (OR=1.7, 95% CI: 1.1, 2.5) and high parity mothers (OR=1.9, 95% CI: 1.1, 3.1). Poor housing, smoking and low social status were also associated with short interpregnancy interval CONCLUSION: Short interpregnancy intervals may be a marker for women at risk and these risk factors differ among populations. They also appear to be a result of choice (e.g. in older women). Biological factors also play a significant role in determining short interpregnancy intervals. PMID- 11380290 TI - Psychological distress before and after prenatal invasive karyotyping. AB - BACKGROUND: The aim was to evaluate the psychological and physical impact on women submitting to early amniocentesis or chorionic villus sampling and their preferences concerning fetal invasive procedures in a forthcoming pregnancy. METHOD: Fifty-one women undergoing early amniocentesis or chorionic villus sampling at 10-13 weeks gestation participated. The women completed questionnaires including the Hospital Anxiety and Depression Scale on three occasions: before, immediately after, and one week after the invasive procedure. RESULTS: The women were more concerned about a spontaneous abortion and waiting for the result than about a possible unreliable result. A majority of the women did not worry about the risk of fetal injury induced by the procedure. More women in the chorionic villus sampling group found the invasive procedure to be physically straining. Two-thirds of the women experienced no or minor psychological strain but one-third experienced major strain. Approximately 20% were classified as doubtful cases and cases of clinical anxiety. Nearly all women would choose to have an invasive procedure of the same kind in a forthcoming pregnancy. CONCLUSIONS: The psychological and physical impact on women undergoing invasive procedures for fetal karyotyping does not constitute a major clinical problem. However, a certain number of women report distress and anxiety which should be attended to by professionals involved in fetal invasive procedures. These women may benefit from more support. PMID- 11380291 TI - Insulin-like growth factor-binding protein-1 in cervical secretion as a predictor of preterm delivery. AB - BACKGROUND: The aim of the study was to evaluate whether the phosphorylated isoforms of insulin-like growth factor-binding protein-1 (IGFBP-1), a protein produced by the decidua, can be detected in cervical secretions of pregnant women with preterm uterine contractions, and whether their presence predicts an increased risk of preterm delivery. METHODS: A prospective analysis of sixty three women who presented with preterm labor but intact fetal membranes at weeks 22-36+6 days of gestation at the Antenatal clinic at the Department of Obstetrics and Gynecology, Helsinki University Central Hospital. Phosphorylated IGFBP-1 (phIGFBP-1) was measured in cervical swab samples obtained at presentation, using an immunoenzymometric assay. The values > or =10 microg/L were considered as positive. In addition, 58 asymptomatic women at the same gestational stage were studied as controls. Multiple logistic regression was applied to control for confounding variables and to obtain adjusted odds ratios. RESULTS: The concentration of phIGFBP-1 in cervical samples ranged from undetectable to 95 microg/L. In 17 of the 63 (27%) women with preterm labor it was > or =10 microg/L. Seven of these 17 (41%) women with a positive phIGFBP-1 result delivered preterm, all before 35 weeks of gestation. Among the women with preterm labor and a negative phIGFBP-1 result, three of the 46 (7%) delivered before 37 weeks of gestation (adjusted OR 24, 95% CI 1.2-487), but all after 35 weeks of gestation. In the asymptomatic control population three out of 58 (5%) women had a positive cervical phIGFBP-1 test result but none delivered preterm. Among the controls with a negative cervical phIGFBP-1 test result (55 of 58, 95%), one woman delivered preterm (1 of 55, 2%). CONCLUSIONS: Pregnant women who are in preterm labor with intact fetal membranes and who have a positive phIGFBP-1 test result in cervical secretion have an increased risk of preterm delivery. PMID- 11380292 TI - National survey (Sweden) of routines for intrapartum fetal surveillance. PMID- 11380293 TI - Changes in serum vascular endothelial growth factor following initiation of estrogen replacement after hysterectomy and oophorectomy. AB - BACKGROUND: Vascular endothelial growth factor has been shown to play an important role in preservation and restoration of endothelial integrity. Similar effects on endothelial function have been described with estrogen replacement. This poses the question whether some of the mechanisms ascribed to estrogen are in fact vascular endothelial growth factor mediated. The objective of this study was to examine the effect of continuous transdermal estrogen replacement on serum levels of secreted vascular endothelial growth factor in women following hysterectomy and oophorectomy. METHODS: In a nonrandomized, prospective study, 40 hysterectomized and oophorectomized women were treated with transdermal estrogen. Their serum vascular endothelial growth factor levels were measured before and six months following estrogen treatment, using a commercially available ELISA kit. RESULTS: The mean serum vascular endothelial growth factor level fell from 328 ng/ml (s.d. 164) before treatment to 285 ng/ml (s.d. 124) following six months of estrogen replacement. This decrease was statistically significant (p<0.03). CONCLUSION: Transdermal estrogen replacement in hysterectomized and oophorectomized women appears to be associated with a significant reduction in secreted serum vascular endothelial growth factor. However, the effect of hysterectomy and oophorectomy per se needs to be explained in appropriately designed trials. PMID- 11380294 TI - Preoperative treatment of anemic women with epoetin beta. AB - AIM: To compare the effects of preoperative treatment with recombinant human erythropoietin (EPO) and iron with iron only on hemoglobin levels (Hb) in anemic women prior to hysterectomy. METHODS: The study was an open-labelled randomized parallel study. Fifteen women scheduled for hysterectomy due to uterine myoma were given oral iron and EPO (NeoRecormon) 4 weeks prior to surgery (group I) and 16 women were given oral iron only (group II). RESULTS: Group I showed a significantly greater increase in mean Hb during the pre-surgery study period compared with group II (p=0.007). Two weeks postoperatively, however, there was no significant difference in mean Hb between the two groups. CONCLUSION: We found that a significantly greater increase in Hb was achieved with iron in combination with EPO, although in most cases iron only seemed to be as efficacious as iron+EPO in correcting anemia in myoma patients pre-operatively. PMID- 11380295 TI - Manual vacuum aspiration, a safe and effective alternative in early pregnancy termination. AB - BACKGROUND AND OBJECTIVE: Vacuum aspiration has become standard surgical procedure for safe early pregnancy termination. Most of these operations are performed in the operating theater using suction curettage and an electric vacuum pump. MVA (manual vacuum aspiration) is an alternative that is well suited for use as a clinical procedure, which could have advantages both for the patient and the health care system. In order to compare conventional VA and MVA in a Swedish setting, a randomized study was undertaken. METHODS: Two hundred women requesting abortion in early pregnancy, gestational age less than 56 days, and choosing surgical termination, were randomized to VA or MVA. Main study outcome was frequency of complete abortion but also other variables were recorded. RESULTS: There were no significant differences between the two groups regarding background characteristics. Altogether 91 MVA and 88 VA operations were performed. There was no significant difference in frequency of complete abortion; two patients in each group subsequently needed re-curettage because of incomplete evacuation. No case of ongoing pregnancy occurred. Two patients in each group received treatment for endometritis. No other complications were recorded. CONCLUSION: This study indicates that MVA is effective in emptying the uterine cavity, on par with the standard vacuum aspiration. The rate of complications with MVA was on the same low level as with conventional VA. PMID- 11380296 TI - Complications of induced abortion and miscarriage in three African countries: a hospital-based study among WHO collaborating centers. AB - OBJECTIVE: The aim of this study was to describe two of the outcomes of pregnancy, induced abortion and miscarriage, in three African countries. Major maternal risk factors were also evaluated. METHODS: The study was prospective and based on the medical files of all 1,957 women admitted to participating health care structures. RESULTS: Overall, 988 women were admitted for complications of miscarriage, and 969 for complications of induced abortion. Gestational age was lower in women with miscarriages (p<0.002). The level of use of contraceptive methods ((p<0.003) and educational level ((p<0.005) were lower in women who had had an induced abortion. In our study, 26 maternal deaths were recorded, 22 of which were associated with induced abortion. Infection was the most important risk factor for death (OR=4.8; 1.9-12.4). CONCLUSION: Maternal deaths related to abortion complications often occurred shortly after hospital admission and with signs of sepsis. This demonstrates the importance of effective emergency services. Unfortunately, hospital-based studies alone cannot assess all maternal death risk factors, especially those for maternal death related to induced abortion complications. It is therefore important to determine what happened to the woman before hospital admission and during her stay in hospital. Combinations of qualitative and quantitative methods could be used to increase our understanding of this problem and to help us to solve it. PMID- 11380297 TI - Prospective, randomized study to evaluate the success rates using hCG, vaginal progesterone or a combination of both for luteal phase support. AB - BACKGROUND: A prospective study was done to compare the efficacy of luteal phase support (LPS) using either three times hCG (group I, n=77), hCG on the day of embryo transfer (ET) in combination with daily vaginal progesterone (group II, n=62) or vaginal progesterone only (group III, n=70). METHOD: All patients were treated using the long luteal protocol for controlled ovarian stimulation in an IVF (in vitro fertilization) cycle. Patients were randomized to one of these groups when estradiol was <2500 pg/ml and less than 12 oocytes were retrieved (low risk groups). If estradiol was > or = 2500 pg/ml and/or at least 12 oocytes were retrieved (high risk groups), patients were randomized to receive either hCG in combination with daily vaginal progesterone (group IV, n=83) or progesterone only (group V, n=121). For vaginal progesterone Utrogest was used (three times daily two capsules containing 100 mg progesterone, 600 mg/d). RESULTS: Demographic data were comparable within the high risk and low risk groups. However, for unknown reasons the fertilization rate was significantly higher in group V (48%) compared to group IV (40%) (p<0.05), leading to a significantly higher cumulative embryo score. There were no statistically significant differences with regard to the main outcome parameter, the clinical ongoing pregnancy rate in the low risk groups (14.3%, 14.5%, 11.4%) and the high risk groups (21.0%, 21.5%), respectively. Using a standardized discomfort scale, there were more complaints towards the end of the luteal phase in the groups receiving hCG only or an additional injection of hCG, when compared to the progesterone only groups. CONCLUSION: Progesterone only for luteal phase support leads to the same clinical ongoing pregnancy rate as hCG, but has no impact on the comfort of the patient. PMID- 11380298 TI - Preoperative CA-125 level as a predictor of non optimal cytoreduction of advanced epithelial ovarian cancer. AB - BACKGROUND: Suboptimal cytoreduction of advanced ovarian cancer is related to initial tumor bulk which correlates with CA125 level. METHODS: Retrospective record study of 40 patients with stage III ovarian cancer. The ability of a CA125 threshold level of 500 U/mL to predict suboptimal cytoreduction was determined. RESULTS: Twenty-four (60%) of the patients were optimally cytoreduced. At the CA125 cut off level of 500 U/mL the sensitivity for predicting suboptimal debulking was 62% and specificity was 83%. Above a CA 125 level of 1500 U/mL none of the patients were optimally cytoreduced. CONCLUSIONS: More data are needed to determine the CA125 cut off level at which the standard approach of initial laparotomy and cytoreduction may be modified. PMID- 11380299 TI - Cervico-isthmic pregnancy: a case report, critical reappraisal of the diagnostic criteria, and reassessment of the outcome. PMID- 11380300 TI - Endometrial adenocarcinoma arising during estrogenic treatment 17 years after total abdominal hysterectomy and bilateral salpingo-oophorectomy: a case report. PMID- 11380301 TI - Have the times come for early intervention in psychosis? PMID- 11380302 TI - Early detection and intervention in first-episode schizophrenia: a critical review. AB - OBJECTIVE: To review the literature on early intervention in psychosis and to evaluate relevant studies. METHOD: Early intervention was defined as intervention in the prodromal phase (primary prevention) and intervention after the onset of psychosis, i.e. shortening of duration of untreated psychosis (DUP) (secondary prevention). RESULTS: We found few studies aimed at early intervention, but many papers discussing the idea at a more general level. We identified no studies that prove that intervention in the prodromal phase is possible without a high risk for treating false positives. We identified some studies aimed at reducing DUP, but the results are ambiguous and, until now, no follow-up data showing a positive effect on prognosis have been presented. CONCLUSION: Early intervention in psychosis is a difficult and important challenge for the psychiatric health services. At the time being reduction of DUP seems to be the most promising strategy. Intervention in the prodromal phase is more ethically and conceptually problematic. PMID- 11380303 TI - Reasoning about the optimal duration of prophylactic antipsychotic medication in schizophrenia: evidence and arguments. AB - OBJECTIVE: To review evidence-based literature regarding the necessary duration of antipsychotic relapse prevention in schizophrenia and related psychoses. METHOD: A computerized search was performed on Medline, Embase Psychiatry and PsycLIT which covered the period 1974-99. We also used cross-references. RESULTS: Although schizophrenia refers mainly to an intrinsic biological vulnerability, only maintenance studies with a follow-up of 2 years at most are available. Relapses appear unpredictable and occur even after long-term successful remission during antipsychotic treatment. CONCLUSION: Since rehabilitation efforts have effects only after long-term endeavours, antipsychotic relapse prevention should be maintained for long periods. It is reasonable to treat patients suffering from schizophrenia and related psychoses for longer periods than indicated by the current guidelines. PMID- 11380304 TI - Predictors and outcome characteristics associated with suicidal behaviour in early psychosis: a two-year follow-up of first-admitted subjects. AB - OBJECTIVE: To assess the baseline characteristics associated with a greater risk of suicidal behaviour (suicide and parasuicide) over the 2 years following a first admission for psychosis, and the associations between suicidality and outcome. METHOD: First-admitted subjects with psychosis (n=65) were assessed at 6 monthly intervals over a 2-year follow-up period. RESULTS: Over this period, 11.3% of the patients displayed suicidal behaviour. Baseline predictors of suicidal behaviour were a lifetime history of parasuicide before first admission (OR=5.9, 95% CI 1.5-23.4), lower Positive And Negative Symptom Scale positive subscores (OR=0.8, 95% CI 0.6-0.97) and a longer duration of first admission (OR=1.1, 95% CI 1-1.2). Subjects with suicidal behaviour presented with a longer duration of psychotic symptoms (OR=1.1, 95% CI 1.02-1.2) and a greater risk of being readmitted (OR=4.6, 95%CI 1.1-19.1). Subjects with substance misuse over the follow-up period were seven times (95%CI 1.3-39) more likely to engage in suicidal behaviour. CONCLUSION: Subjects with a previous history of parasuicide, with a deteriorating clinical course, or with substance misuse are at increased risk of suicidal behaviour in the 2 years after the onset of a first psychotic episode. PMID- 11380305 TI - Impact of phase-specific treatment of first episode of psychosis on Wisconsin Quality of Life Index (client version). AB - OBJECTIVE: The objective of this study was to assess the impact of a phase specific community-focused treatment program on different dimensions of self reported quality of life in a representative sample of first episode psychosis patients. METHOD: Data were collected on patients presenting with a first episode of psychosis on the Wisconsin Quality of Life Index (client version), positive and negative symptoms, and demographic and clinical variables at baseline following clinical stabilization and at 1 year. RESULTS: Complete data on a representative sample of 41 patients showed a significant improvement in most dimensions of the WQOL at 1 year; these changes were generally independent of changes in symptoms and there were no significant differences in the magnitude of improvement in QOL between those with DUP < or >6 months. CONCLUSION: Patients with a first episode of predominantly schizophrenia spectrum psychosis show a highly significant improvement in subjectively assessed quality of life following a year of phase-specific comprehensive treatment. PMID- 11380306 TI - A randomized, controlled trial of a brief interventional package for schizophrenic out-patients. AB - OBJECTIVE: To evaluate the impact on outcome of a simple educational intervention in schizophrenic patients at risk of relapse. METHOD: At discharge, 114 schizophrenic patients with at least one previous episode were assigned randomly to a simple educational intervention which had no resource implications, or standard care. RESULTS: The intervention failed to improve outcome. While insight and treatment attitudes improved, suicidal ideation increased. Systematic management of treatment-emergent adverse effects offered no benefits, although incapacitation from extrapyramidal side-effects at discharge predicted relapse. CONCLUSION: There are limits to which psychoeducational interventions can be simplified without loss of effectiveness in terms of relapse prevention in schizophrenia. Enhanced insight may be associated with increased suicidal ideation. PMID- 11380307 TI - A comparison of needs of patients with schizophrenia in five European countries: the EPSILON Study. AB - OBJECTIVE: To compare the occurrence of needs of patients with schizophrenia in Amsterdam, Copenhagen, London, Santander and Verona. METHOD: Patients with schizophrenia were interviewed with the Camberwell Assessment of Need. The mean numbers of total, met and unmet needs were compared across the sites. Ratings for individual domains were also examined. Means were adjusted, using multiple regression analysis, to control for patient differences between sites. RESULTS: The highest levels of unmet need were in Amsterdam and London. The number of met needs were similar. Adjusting for patient differences reduced the estimated number of unmet needs and total needs in Copenhagen, but made little difference elsewhere. There were many site differences for individual domains. CONCLUSION: Needs vary between different countries. Unmet needs are greater in large urban areas like London and Amsterdam. Differences in the pattern of individual needs may be partly explained in terms of service provision. PMID- 11380308 TI - A comparison of community-dwelling older schizophrenia patients by residential status. AB - OBJECTIVE: Differences in residential status may contribute to the diversity of findings observed in community-based samples of patients with schizophrenia. We compared older out-patients living independently with those in assisted-care facilities. METHOD: Two hundred and fifty-one out-patients with schizophrenia or schizoaffective disorder, aged 40-97 years, who had been referred to our Intervention Research Center were examined. RESULTS: Assisted living status was associated with an earlier age at onset of illness, longer illness duration, lower probability of having ever been married, more severe negative symptoms, worse cognitive impairment, and a poorer health-related quality of wellbeing. Independent living and assisted-care patients had similar levels of positive and depressive symptoms, and were on comparable doses of neuroleptic medication. Significant 'predictors' of residential status were marital status, cognitive impairment and quality of wellbeing. CONCLUSION: Among schizophrenia out patients, one needs to consider residential status in evaluating heterogeneity in cognitive performance or quality of wellbeing. PMID- 11380309 TI - Subject and observer-rated quality of life in schizophrenia. AB - OBJECTIVE: We aimed to explore the relationship between objectively rated quality of life and subjective measures of social functioning and life satisfaction. METHOD: Participants of the Schizophrenia Care and Assessment Program (SCAP) study at Dandenong in Australia were included in this analysis. Subjective ratings of several domains of social functioning and life satisfaction were taken from the SCAP instrument and comparisons made with data from the Quality of Life scale rated by research staff as well as several psychopathology measures. RESULTS: Subjectively reported life satisfaction was not related to positive or negative symptoms of schizophrenia but did correlate with depressive symptoms. Quality of Life scale measures correlated with negative symptoms on most domains. There was very limited overlap in domain items between the life satisfaction and quality of life measures. CONCLUSION: Life satisfaction and objectively rated quality of life are not closely related and appear to have different determinants in patients with schizophrenia. PMID- 11380310 TI - Cognitive behaviour therapy with coping training for persistent auditory hallucinations in schizophrenia: a naturalistic follow-up study of the durability of effects. AB - OBJECTIVE: To investigate the durability of positive effects of cognitive behaviour therapy (CBT) with coping training on psychotic symptoms and social functioning. METHOD: Forty patients with schizophrenia or related psychotic disorders and refractory auditory hallucinations were given CBT and coping training in an integrated single family treatment programme. In a naturalistic study patients were followed after 2 and 4 years since the start of treatment. RESULTS: The treatment improved overall burden of 'hearing voices', with a generalization into daily functioning. Improvement with regard to fear, loss of control, disturbance of thought and interference with thinking was sustained by 60% of the patients while one-third improved further. Complete disappearance of hallucinations occurred in 18% of the patients. CONCLUSION: CBT with coping training can improve both overall symptomatology and quality of life, even over longer periods of time, but a status of persistent disablement indicates a continuing need for mental health care. PMID- 11380311 TI - Lithium intoxication secondary to unrecognized pontine haemorrhage. AB - Lithium prophylaxis carries a substantial risk of medical complications, especially in the case of concomitant medical conditions. We describe a patient with unrecognized cerebrovascular haemorrhage, admitted to hospital due to lithium intoxication. PMID- 11380312 TI - Review article: the genetics of inflammatory bowel disease. AB - Recent epidemiological, clinical and molecular studies have provided strong evidence that inherited predisposition is important in the pathogenesis of chronic inflammatory bowel diseases. The model most consistent with the epidemiological data suggests that Crohn's disease and ulcerative colitis are related polygenic diseases, sharing some but not all susceptibility genes. Investigators throughout the world have applied the complementary techniques of genome-wide scanning and candidate gene analysis. Four areas of linkage have been widely replicated on chromosomes 16 (IBD1), 12 (IBD2), 6 (IBD3-the HLA region), and most recently on chromosome 14. Fine mapping of these regions is underway. Of the 'positional' candidate genes, most attention has centred on the genes of the major histocompatibility complex. Genes within this region may determine disease susceptibility, behaviour, complications and response to therapy. Hope continues that studies of inflammatory bowel disease genetics will provide fresh insight into disease pathogenesis and soon deliver clinical applications. PMID- 11380313 TI - Review article: the therapy of constipation. AB - Constipation is a common symptom that may be idiopathic or due to various identifiable disease processes. Laxatives are agents that add bulk to intestinal contents, that retain water within the bowel lumen by virtue of osmotic effects, or that stimulate intestinal secretion or motility, thereby increasing the frequency and ease of defecation. Drugs which improve constipation by stimulating gastrointestinal motility by direct actions on the enteric nervous system are under development. Other modalities used to treat constipation include biofeedback and surgery. Laxatives and lavage solutions are also used for colon preparation and evacuation of the bowels after toxic ingestions. PMID- 11380314 TI - Review article: acid suppression in the management of gastro-oesophageal reflux disease--an appraisal of treatment options in primary care. AB - Gastro-oesophageal reflux disease (GERD) is one of the most common conditions presenting to the primary care physician. Despite progress in understanding and treatment of the disease, strategies for capitalizing on these advances are less well developed. In many practices, H2-receptor antagonists still remain the most widely prescribed treatment for GERD, despite the availability of the more effective acid-suppressant proton pump inhibitors. This review examines the relative efficacies of acid-suppressant drugs in minimizing oesophageal acid exposure and outlines the evidence for the superiority of proton pump inhibitors over standard-dose H(2)-antagonists in symptom relief, erosion healing and prevention of relapse in GERD. Current prescribing patterns and considerations for the general practitioner are also examined. The availability and impact of over-the-counter H(2)-antagonists on the treatment of GERD and their relative cost-effectiveness vs. proton pump inhibitors are also addressed. A hierarchy of drug efficacy (full-dose proton pump inhibitor > half-dose proton pump inhibitor > high-dose H(2)-antagonist > standard-dose H(2)-antagonist or prokinetic) applies in principle to all GERD patients, (with or without oesophagitis). The most effective initial therapy for GERD is also likely to be the most cost effective one, if treatment failure leads to higher utilization of medical resources. The application of these recommendations to the management of non endoscoped GERD, endoscopy-negative GERD and low-grade oesophagitis as well as higher grade oesophagitis is also reviewed. PMID- 11380315 TI - Guidelines for adults on self-medication for the treatment of acute diarrhoea. AB - Acute uncomplicated diarrhoea is commonly treated by self-medication. Guidelines for treatment exist, but are inconsistent, sometimes contradictory, and often owe more to dogma than evidence. An ad hoc multidisciplinary group has reviewed the literature to determine best practice. In general it is recognized that treatment of acute episodes relieves discomfort and social dysfunction. There is no evidence that it prolongs the illness. Self-medication in otherwise healthy adults is safe. Oral loperamide is the treatment of choice. Older anti-diarrhoeal drugs are also effective in the relief of symptoms but carry the risk of unwanted adverse effects. Oral rehydration solutions do not relieve diarrhoea, and confer no added benefit for adults who can maintain their fluid intake. Probiotic agents are, at present, limited in efficacy and availability. Antimicrobial drugs, available without prescription in some countries, are not generally appropriate for self-medication, except for travellers on the basis of medical advice prior to departure. Medical intervention is recommended for the management of acute diarrhoea in the frail, the elderly (> 75 years), persons with concurrent chronic disease, and children. Medical intervention is also required when there is no abatement of the symptoms after 48 h, or when there is evidence of deterioration such as dehydration, abdominal distension, or the onset of dysentery (pyrexia > 38.5 degrees C and/or bloody stools). PMID- 11380316 TI - Abnormal bone turnover in long-standing Crohn's disease in remission. AB - BACKGROUND: A high prevalence of osteoporosis is found in patients with Crohn's disease. The pathogenesis of this condition seems to be multifactorial and its pathophysiology is still not completely understood. AIM: To elucidate the pathophysiology of osteopenia in quiescent Crohn's disease. METHODS: Bone turnover was studied in 26 patients (13 males and 13 females) with long-standing quiescent Crohn's disease and small bowel involvement. Bone mineral density was assessed by dual energy X-ray absorptiometry. Biochemical markers for bone formation (osteocalcin and bone-specific alkaline phosphatase) and for bone resorption (deoxypyridinoline and collagen type I C-terminal crosslinks) were measured. Urinary calcium excretion was determined. RESULTS: Markers for bone formation were significantly lower in patients than in controls (osteocalcin: P= 0.027, bone-specific alkaline phosphatase: P < 0.001), but both bone resorption markers were not significantly different. Urine calcium excretion was significantly decreased in patients (P=0.002) compared to controls. Bone mineral density of the lumbar spine was significantly and inversely correlated with bone specific alkaline phosphatase and collagen type I C-terminal crosslinks. CONCLUSIONS: Bone turnover in long-standing Crohn's disease in clinical remission is characterized by suppressed bone formation and normal bone resorption. Urine calcium excretion is decreased. Hence, interventions and therapy should be directed towards the improvement of bone formation. PMID- 11380318 TI - The effect of Helicobacter pylori eradication therapy on dyspepsia symptoms in industrial workers in Japan. AB - BACKGROUND: The relationship between Helicobacter pylori infection and non-ulcer dyspepsia is still controversial. The potential benefits and risks of the treatment could depend on local conditions, such as the prevalence of the infection and the local rates of gastric cancer. AIM: To evaluate the effects of H. pylori eradication therapy on non-ulcer dyspepsia symptoms in industrial workers in Japan. METHODS: A total of 615 employees of an industrial corporation were examined for H. pylori infection and symptom scores; 215 H. pylori-positive non-ulcer dyspepsia cases underwent eradication therapy. Symptom scores were also analysed 12 months after the eradication therapy. Serum pepsinogen A and pepsinogen C levels were analysed and chronic atrophic gastritis was serologically diagnosed on the basis of the criteria of a pepsinogen A < 70 ng/mL and pepsinogen A : pepsinogen C ratio < 3.0. RESULTS: The symptom score improved significantly in the cured cases, but not in the non-cured cases. The effect of the cure of H. pylori infection on symptoms was analysed according to the serological diagnosis of chronic atrophic gastritis. In both groups, cases with atrophic gastritis and cases with chronic gastritis only, the cure of infection was effective in improving symptoms. CONCLUSION: The cure of H. pylori infection was effective in reducing non-ulcer dyspepsia symptoms in industrial workers in Japan. PMID- 11380317 TI - Pharmacodynamic effects and kinetic disposition of rabeprazole in relation to CYP2C19 genotypes. AB - BACKGROUND: S-mephenytoin 4'-hydroxylase (CYP2C19) catalyses the metabolism of rabeprazole to some extent. Based on the metabolic and pharmacokinetic differences among other proton pump inhibitors such as omeprazole, lansoprazole and pantoprazole, rabeprazole appears to be the least affected proton pump inhibitor by the CYP2C19-related genetic polymorphism. AIM: To determine whether the pharmacodynamic effects of rabeprazole on intragastric pH and serum gastrin levels, and its pharmacokinetics depend on the CYP2C19 genotype status. METHODS: Eighteen healthy subjects, whose CYP2C19 genotype status was previously determined, participated in the study. They consisted of six each of homozygous extensive metabolisers (homo EMs), heterozygous extensive metabolisers (hetero EMs), and poor metabolisers (PMs). Helicobacter pylori status was determined by serology. After a single oral dose of 10 mg or 20 mg rabeprazole or water only (baseline data), intragastric pH values were monitored for 24 h. Plasma levels of rabeprazole and serum gastrin were also measured for 24 h post-dose. RESULTS: Five homo EM, six hetero EM and four PM subjects were H. pylori-negative. After rabeprazole administration, significant differences in intragastric mean pH values, serum gastrin AUC(0-24) and plasma levels of rabeprazole were observed among the three different genotype groups. CONCLUSION: The pharmacodynamic effects of rabeprazole and its pharmacokinetics depend on the CYP2C19 genotype status. PMID- 11380319 TI - Increased gastric acid secretion after Helicobacter pylori eradication may be a factor for developing reflux oesophagitis. AB - BACKGROUND: The role of acid secretion in reflux oesophagitis which may develop after H. pylori eradication is not well known. AIM: To investigate the participation of altered gastric acid secretion and the presence of hiatal hernia in the development of reflux oesophagitis after eradication therapy for H. pylori. SUBJECTS AND METHODS: A total of 105 patients with H. pylori infection, but without reflux oesophagitis at the time of eradication therapy, were followed prospectively for 7 months after the clearance of this microorganism. Gastric acid secretion was assessed by endoscopic gastrin test, and the presence of hiatal hernia by endoscopy. RESULTS: Reflux oesophagitis developed in 11 out of 105 (10.5%) patients when examined at 7 months after the eradication therapy. The incidence was correlated significantly with the increase in gastric acid secretion after the eradication of H. pylori, and was significantly higher in the patients with hiatal hernia (20%) than in those without it (0%). CONCLUSIONS: Increased acid secretion after H. pylori eradication is an important risk factor of reflux oesophagitis, especially in patients with hiatal hernia. PMID- 11380320 TI - Occurrence and relapse of bleeding from duodenal ulcer: respective roles of acid secretion and Helicobacter pylori infection. AB - BACKGROUND: Helicobacter pylori infection, gastric acid hypersecretion and NSAID consumption may cause peptic ulcer. AIM: To investigate the respective roles of H. pylori and acid secretion in bleeding duodenal ulcer. PATIENTS AND METHODS: A total of 99 duodenal ulcer patients were referred for evaluation of acid secretion: seven with Zollinger-Ellison Syndrome; 14 with hypersecretory duodenal ulcer, defined by the coexistence of elevated basal acid output and pentagastrin acid output; and 78 duodenal ulcer patients with normal acid output. All non Zollinger-Ellison Syndrome patients were H. pylori-positive and cured of infection. All patients were followed-up for a 36-month period, to assess the occurrence of bleeding episodes. RESULTS: Twenty-nine patients had at least one bleeding episode in the 4 years before the study. Bleeding was more frequent in males and in patients on NSAIDs. The mean basal acid output was not higher among bleeders. In the 21 patients (14 hypersecretory duodenal ulcer, seven Zollinger Ellison Syndrome) with basal acid output > 10 meg/h and pentagastrin acid output > 44.5 meg/h, the risk of bleeding was higher (OR 6.5; 95% CI: 2-21). In the follow-up period, three out of 83 (3.3%) non-Zollinger-Ellison Syndrome patients had a H. pylori-negative duodenal ulcer with bleeding. The risk of bleeding after H. pylori cure was not higher in hypersecretory duodenal ulcer patients (P > 0.3), nor among patients with previous bleeding episodes (P > 0.2). CONCLUSIONS: In H. pylori-positive duodenal ulcer patients, the coexistence of elevated basal acid output and pentagastrin acid output leads to a sixfold increase in the risk of bleeding. After H. pylori cure, gastric acid hypersecretion is not a risk factor for bleeding. However, duodenal ulcer recurrence with bleeding may occasionally occur in patients cured of H. pylori, even if acid output is normal. PMID- 11380321 TI - Anti-inflammatory and tissue-protectant drug effects: results from a randomized placebo-controlled trial of gastritis patients at high risk for gastric cancer. AB - BACKGROUND: The inflammatory process involving Helicobacter pylori-associated gastritis is thought to lead to epithelial damage and contribute to the development of gastric cancer. Evidence exists from animal and in vitro studies suggesting that tetracyclines have both anti-inflammatory and tissue-protectant effects unrelated to their antimicrobial activity. We attempted to modulate components of H. pylori's inflammatory process by: (i) eliminating the infection; (ii) using tetracycline to alter the host's reaction to the infection without reducing the bacterial load; and (iii) using calcium to counteract the effect of excessive dietary salt. METHODS: We conducted a 16-week placebo-controlled clinical trial with 374 H. pylori-associated gastritis patients randomly assigned to one of five groups: (1) triple therapy consisting of metronidazole, amoxicillin and bismuth subsalicylate for 2 weeks, followed by bismuth alone for 14 weeks; (2) calcium carbonate; (3) triple therapy and calcium carbonate; (4) tetracycline; or (5) placebo. RESULTS: Subjects in the tetracycline and triple therapy groups, but not the calcium carbonate only group, showed a reduction in inflammation and epithelial damage vs. those in the placebo group, independent of a change in H. pylori density and other factors. Our results also indicate that epithelial damage may be affected by mechanisms independent of H. pylori density or inflammation. CONCLUSION: The results are consistent with the hypothesis that tetracycline can decrease inflammation independent of a reduction in the bacterial load. More research is needed to investigate mechanisms leading to epithelial damage which are independent of H. pylori density and inflammation. PMID- 11380323 TI - Fish oil (Eicosapen) is less effective than metronidazole, in combination with pantoprazole and clarithromycin, for Helicobacter pylori eradication. AB - BACKGROUND: In vitro omega-3-fatty acids (Eicosapen) are bacteriostatic to Helicobacter pylori and have a variety of immuno-modulating effects. AIM: To investigate the efficacy and tolerability of eicosapen (E) as an antibiotic sparing component of a triple H. pylori eradication regimen in non-ulcer dyspepsia patients in a randomized, double-blind trial. METHODS: Non-ulcer dyspepsia patients (n=199), with a normal upper endoscopy and a positive (13)C urea breath test (UBT) were randomly assigned to either pantoprazole, clarithromycin and metronidazole (PCM) or pantoprazole, clarithromycin and eicosapen (PCE) for 7 days. Four weeks after treatment, H. pylori eradication was determined by UBT. Symptoms were followed up to 16 months. RESULTS: In the intention-to-treat population, PCM eradicated infection in 78% but PCE was successful in only 34% (P < 0.001). Symptomatic improvement occurred in both groups, and was not related to H. pylori eradication. CONCLUSION: Eicosapen is unlikely to be useful in H. pylori eradication regimens. PMID- 11380322 TI - Three-day lansoprazole quadruple therapy for Helicobacter pylori-positive duodenal ulcers: a randomized controlled study. AB - AIM: To compare the efficacy and tolerability of a 3-day quadruple therapy with a standard 7-day triple therapy in eradicating Helicobacter pylori infection and healing duodenal ulcers. METHODS: Patients with H. pylori-positive duodenal ulcers were randomized to receive either lansoprazole 30 mg, clarithromycin 500 mg, and metronidazole 400 mg twice daily for 7 days (LCM-7) or lansoprazole 30 mg, clarithromycin 500 mg, metronidazole 400 mg, and bismuth subcitrate 240 mg twice daily for 3 days (LCMB-3). No pre- or post-treatment acid suppression was used. Follow-up endoscopy was performed at week 6. RESULTS: A total of 118 patients were recruited. Sixty patients in the LCM-7 group and 53 patients in the LCMB-3 group returned for endoscopy. Intention-to-treat eradication rates were 87% and 86% (P=0.94) and per protocol eradication rates were 87% and 94% (P=0.29) in the LCM-7 and LCMB-3 groups, respectively. Per protocol and intention-to-treat ulcer healing rates were 98% and 98% in LCM-7 and 100% and 91% in LCMB-3, respectively. There were no significant differences in efficacy in relation to the initial metronidazole and clarithromycin susceptibility. Significant reduction in the duration of side-effects was found in the LCMB-3 group. CONCLUSION: The 3-day quadruple therapy is highly effective, better tolerated and can be considered as a first-line therapy in duodenal ulcer management. PMID- 11380324 TI - Bolus somatostatin but not octreotide reduces hepatic sinusoidal pressure by a NO independent mechanism in chronic liver disease. AB - BACKGROUND: Evidence exists that somatostatin and octreotide might have different effects on hepatic haemodynamics. AIM: The investigation of the effects of somatostatin and its octapeptide analogue, octreotide, on sinusoidal pressure measured by the wedged hepatic venous pressure in patients with cirrhosis or chronic hepatitis and the correlation with the levels of hepatic vein NO. METHODS: Patients were randomly assigned to receive an injection of either 250 microg somatostatin (n=14: cirrhosis six, chronic hepatitis eight) or an injection of 125 microg octreotide (n=19: cirrhosis nine, chronic hepatitis 10) during hepatic vein catheterization. Baseline wedged hepatic venous pressure was measured, followed by measurements at 2, 5, 10 and 15 min after the injection of the drug. Nitrites/nitrates of the hepatic vein were measured before the injection and after 15 min. RESULTS: Both agents showed a similar qualitative but a different quantitative haemodynamic profile. No change in the wedged hepatic venous pressure was observed during the first 2 min after the injection of both drugs. This was followed by a decrease: 18% at 5 min (N.S.), 23% at 10 min (P < 0.01) and 24% at 15 min (P < 0.01) for somatostatin. Octreotide induced a relatively smaller decrease in the wedged hepatic venous pressure: 8% at 5 min (N.S.), 20% at 10 min (P < 0.01) and 16% at 15 min (N.S.). Further analysis of the sub-groups of cirrhotic and chronic hepatitis patients revealed a different effect. In the sub-group of cirrhotic patients, somatostatin caused a maximum decrease of 34% at 15 min post-injection (P < 0.01), but octreotide failed to produce a significant change on the wedged hepatic venous pressure. In contrast, no change was observed in chronic hepatitis patients with either drug. No change in the hepatic vein concentration of NO after treatment was observed with either somatostatin or octreotide. Moreover, no correlation of the levels of NO with the wedged hepatic venous pressure values was found. CONCLUSIONS: This study shows that somatostatin is more effective than octreotide in acutely reducing the wedged hepatic venous pressure after bolus injection and the observed change is probably mediated by a NO-independent mechanism. PMID- 11380325 TI - Pioglitazone, a specific PPAR-gamma ligand, inhibits aspirin-induced gastric mucosal injury in rats. AB - BACKGROUND: Neutrophils activation and tumour necrosis factor-alpha (TNF-alpha) induction play a critical role in aspirin-induced gastric mucosal injury. Peroxisome proliferator-activated receptor-gamma (PPAR-gamma), a member of the nuclear hormone receptor superfamily, has recently been implicated as a regulator of inflammatory responses. The aim of the present study was to determine whether pioglitazone, a specific PPAR-gamma ligand, can ameliorate aspirin-induced gastric mucosal injury in rats, and whether the agent can inhibit the increase in neutrophil accumulation associated with TNF-alpha expression. METHODS: Aspirin induced injury was produced by the intragastric administration of aspirin (200 mg/kg) and HCl (0.15 N, 8.0 mL/kg). Pioglitazone was given to the rats by gastric intubation 1 h before the aspirin administration. Thiobarbituric acid-reactive substances and tissue-associated myeloperoxidase activity were measured in gastric mucosa as indices of lipid peroxidation and neutrophil infiltration. The gastric concentration of TNF-alpha and the expression of TNF-alpha mRNA was determined by ELISA and reverse transcriptase-polymerase chain reaction. RESULTS: The intragastric administration of acidified aspirin induced hyperemia and haemorrhagic erosions in rat stomachs. The increase in the total gastric erosive area after aspirin administration was significantly inhibited by treatment with pioglitazone in a dose-dependent manner. The increases in thiobarbituric acid reactive substances and myeloperoxidase activity after aspirin administration were both significantly inhibited by pre-treatment with pioglitazone (10 mg/kg). The gastric content of TNF-alpha increased and the expression of TNF-alpha mRNA was up-regulated after aspirin treatment. However, the peak TNF-alpha mRNA expression 1 h after aspirin administration was inhibited by pioglitazone. CONCLUSION: Based on these data, the beneficial effects of pioglitazone on aspirin-induced gastric mucosal injury may be attributed to its anti-inflammatory properties. PMID- 11380326 TI - Involvement of cyclooxygenase-2 in hyperplastic gastritis induced by Helicobacter pylori infection in C57BL/6 mice. AB - BACKGROUND AND AIMS: The hyperplastic changes observed in Helicobacter pylori associated gastritis have been considered to increase the risk of gastric cancer. The aim of this study was to determine whether cyclooxygenase-2 is involved in the hyperplastic changes in mice infected with H. pylori. METHODS: Seven-week old, male C57BL/6 mice (n=40) were inoculated with the Sydney strain of H. pylori. Control mice (n=40) were treated with vehicle only. Half of the infected and control mice were fed an experimental diet containing etodolac (10 mg/kg/day) from 1 week after inoculation until the end of the experiment. The thickness of gastric pits, COX-2 mRNA and protein levels, and prostaglandin E2 (PGE2) levels in the gastric mucosa were determined before and 12, and 24 weeks after inoculation. RESULTS: The thickness of gastric pits, COX-2 mRNA and protein levels, and PGE2 levels were significantly increased at 24 weeks after inoculation of H. pylori compared with the control groups. Treatment with etodolac resulted in significant decreases in PGE2 production and in the thickness of gastric pits in the infected groups at 24 weeks after inoculation. CONCLUSIONS: Our findings suggest that COX-2 is involved in the development of hyperplastic gastritis caused by H. pylori infection via the production of PGE2. PMID- 11380327 TI - Vasectomy. AB - Vasectomy is regarded as the safest method now available for male fertility control. Almost 100 million men worldwide have relied on vasectomy for family planning. This review discusses all currently relevant operative techniques, including no-scalpel vasectomy, complications, possible long-term effects on the testis and epididymis, and diseases for which associations with vasectomy have been suggested, such as arteriosclerosis, autoimmune diseases and cancer of the prostate and testis. Other topics of discussion include the timing of post operative semen analysis, patient noncompliance concerning post-operative controls, persistent cryptozoospermia and transient reappearance of spermatozoa after vasectomy, vasectomy failure and legal aspects. PMID- 11380328 TI - Stepwise regression analysis to study male and female factors impacting on pregnancy rate in an intrauterine insemination programme. AB - The aim of this study was to evaluate the impact of male and female factors on the pregnancy rate in an intrauterine insemination (IUI) programme. Data on 522 cycles were retrospectively studied. All patients 39 years or younger were included in the study where data were available on male and female diagnosis, as well as on ovulation induction methodology. Regression analysis was possible on 495 cycles to study different factors affecting the pregnancy rate per treatment cycle. Logistic regression identified variables which were related to outcome and were subsequently incorporated into a statistical model. The number of follicles was found to have a linear association with the risk ratio (chance) of pregnancy. The age of the woman was also found to have a linear (negative) association with pregnancy. The percentage motility and percentage normal morphology (by strict criteria) of spermatozoa in the fresh ejaculate were the male factors that significantly and independently predicted the outcome. Percentage motility > or = 50 was associated with a risk ratio of pregnancy of 2.95 compared to percentage motility < 50. Percentage normal sperm morphology > 14% was associated with a risk ratio of pregnancy of 1.8 compared to percentage normal morphology < or = 14%. Female patients with idiopathic infertility were divided into three groups according to normal sperm morphology. The pregnancy rate per cycle was 2.63% (1/38) for the P (poor) pattern group (0-4% normal forms), 11.4% (17/149) for the G (good) pattern group (5-14%), and 24% (18/75) for the N (normal) pattern group (> 14% normal forms). A female diagnosis of endometriosis or tubal factor impacted negatively on the probability of pregnancy (risk ratio of 0.17), compared with other female diagnoses. Male and female factors contribute to pregnancy outcome, but the clinician can influence prognosis by increasing the number of follicles, especially in severe male factor cases. PMID- 11380329 TI - Enrichment of high-quality spermatozoa in bovine semen: relative effectiveness of three filtration matrixes. AB - To be practical, any method for improving bull semen must yield a large quantity of motile spermatozoa. Some separation methods based on physical properties, e.g. filtration, chromatography, centrifugation, washing and pooling, have been reported as satisfactory, but generally are not repeatable. Nevertheless, filtration methods appear to allow the attainment of an acceptable number of spermatozoa, thus allowing such a technique to be introduced in the production of standard bovine semen doses for artificial insemination. The aim of this work was to evaluate systematically the relative effects of three filtration matrixes (silica oxide, glass beads or Sephadexatrade mark) on the improvement of whole ejaculate quality. Analysis of the type of matrix and the volume and height of the filtration column was performed. The only characteristic of the columns that appears to influence ejaculate quality after filtering is the matrix volume. While all matrixes produced improvement of semen quality, SephadexTM was better than the other matrixes tested. An explanation for the mechanism of column filtration is proposed. PMID- 11380330 TI - Progressively motile human spermatozoa are well protected against in vitro lipid peroxidation imposed by induced oxidative stress. AB - Semen samples of 24 patients were analysed. Volumes were measured and the numbers of progressively motile (PMS), motile (MS) and nonmotile spermatozoa (NMS) were determined. These 24 samples appeared to show a large variation in motility percentages and numbers. Spermatozoa of these semen samples were isolated from the seminal plasma and exposed to induced radical oxygen stress imposed by iron/ascorbate. Lipid peroxidation (LPO) was quantified as thiobarbituric acid reactive material. The contributions of PMS, MS and NMS were also estimated. It was found that the PMS did not contribute to the formation of lipid peroxides. The cellular radical defence system of PMS may offer them adequate protection against the harsh conditions of radical oxygen stress. Stepwise regression analyses showed that only the population of NMS contributed significantly to the explanation of the variance in LPO production (R2 = 0.56, P < 0.001). Pre existing membrane lipid peroxides were not detected in spermatozoa. It is therefore suggested that LPO takes place only after radical oxygen stress has exhausted the cellular defence system. LPO is not the initial, but one of the later, events leading to the death of spermatozoa. It is concluded that the population of progressively motile spermatozoa in semen samples does not contribute to the production of thiobarbituric acid reactive substances as induced by in vitro radical oxygen stress. PMID- 11380331 TI - Comparison of normal sperm morphology outcomes from two different computer assisted semen analysis systems. AB - By eliminating the human evaluation variable, it was possible to carry out an investigation into the 'true' association between normal sperm morphology outcomes assessed according to the World Health Organization guidelines and strict criteria. Two computer-assisted semen analysis systems were used, IVOS and Mika, to evaluate Diff-Quik and Papanicolaou stained slides. As expected, the mean normal sperm morphology outcomes for the World Health Organization classification evaluations were markedly higher for both the Diff-Quik (mean difference = 40.13%) and the Papanicolaou (mean difference = 32.55%) stained slides. The association between the outcomes were low for the Diff-Quik stained slides (r = 0.379) and poor for the Papanicolaou stained slides (r = 0.110). While the association achieved with the computer-assisted semen analysis systems using Diff-Quik stained slides was comparable to the association between the manual evaluations (r = 0.386), the manual evaluation of Papanicolaou stained slides produced a relatively good association (r = 0.690). Although the numbers were small, the results show the probability of poor class correlations. Approximately 40% of outcomes were incorrectly classed at a 14% (strict criteria) and 50% (World Health Organization guidelines) cut-off point for both staining methods. This study confirms the fundamental differences between the two classification systems. The results also indicate that, of the two stains used, Diff-Quik should be the preferred staining method for computer-assisted sperm morphology evaluations. PMID- 11380332 TI - Expression of aquaporins 7 and 8 in the developing rat testis. AB - Aquaporin (AQP) is a molecule that facilitates water permeability through cell membranes and is widely distributed in the mammalian body. Among members of the AQP family, AQP7 and AQP8 are predominantly expressed in the testis. This paper examined the expression of these two AQPs in the developing rat testis, using northern blot and immunohistochemistry to visualize their roles during the progression of spermatogenesis in the developing testis. In adult rats, expression of AQP7 was localized to spermatids, whereas AQP8 protein was observed in spermatocytes. In the developing testis, transcripts of AQP7 became detectable between 23 and 25 days post-partum, when round spermatids have been reported to appear. On the other hand, AQP8 mRNA was first detected between 13 and 16 days post-partum, consistent with the reported first appearance of spermatocytes (13 14 days). AQP7 and AQP8 proteins were detectable at 28 days post-partum. The results suggest that AQP7 and AQP8 appear in the testis in accordance with the maturation of germ cells, and are involved in spermatogenesis. PMID- 11380351 TI - Non-vascular intervention with real-time CT fluoroscopy. AB - The emerging technology of CT fluoroscopy (CTF) represents the first opportunity for real-time CT guidance in non-vascular intervention. As with any new technology, its efficacy requires validation before widespread application can be advocated. A review of our initial experience is presented with particular attention to room, procedure and fluoroscopy time savings, complication rates and dosimetry. Computed tomography fluoroscopy is useful for pulmonary, pelvic, retroperitoneal and other deep organ lesions that are not easily accessible by other modalities. Computed tomography fluoroscopy decreases procedure time by at least a factor of 2 compared with conventional CT (C-CT) guidance, resulting in improved throughput in a busy interventional CT department. Accurate targeting of small lesions, previously considered inaccessible, can also be achieved with CTF. Exposure to the physician's hands can be reduced to a level that is acceptable to the International Commission on Radiological Protection guidelines. PMID- 11380352 TI - Pain during mammography: implications for breast screening programmes. AB - Pain experienced during mammography can deter women from attending for breast cancer screening. Review of the current literature on pain experienced during mammography reveals three main areas of interest: reports of the frequency of pain, identification of predictors of pain and strategies for responding to pain. Implications of this literature for breast screening programmes include the need for appropriate measurements of pain during mammography that are valid for screening populations, a further understanding of organizational factors involved in screening programmes that may be predictors of pain and for the development of valid strategies for responding to pain within breast screening programmes. PMID- 11380353 TI - Comparison of spiral CT angiography with conventional digital subtraction angiography in the evaluation of renal transplant donors: a pilot study. AB - Conventional digital subtraction renal arteriography (IA-DSA) has been traditionally used as the preoperative imaging modality for assessment of renal vascular anatomy for renal transplant donors. This study evaluates the potential use of spiral CT angiography in replacing IA-DSA in the preoperative assessment of this group of patients. Seven patients underwent both spiral CT angiography and IA-DSA between October 1997 and April 1998. It is concluded that spiral CT angiography can demonstrate the number, length and location of renal arteries and it is suggested that spiral CT angiography can potentially replace IA-DSA in the preoperative assessment of renal donors. PMID- 11380354 TI - Cystic lumbar nerve sheath tumours: MRI features in five patients. AB - Intraspinal cystic lumbar nerve sheath tumours constitute an uncommon subset of tumours with distinct clinico-biological behaviour. The MRI findings in five such cases are presented. Four of these were cystic schwannomas and one was a cystic neurofibroma. The pathology, MRI findings and the differential diagnosis of these rare tumours are analysed. PMID- 11380355 TI - Magnetic resonance imaging signal characteristics in hydatid cysts. AB - The MRI features of hydatid cysts were retrospectively studied in 12 patients to look for specific signal characteristics. Twelve patients in the series included eight with abdominal, two with abdominal-pelvic, one with a sacral and one with a lung hydatid cyst. The T1-, T2- and proton density (PD)-weighted images of spin echo sequence were used for imaging. The signal characteristics of the hydatid parent cyst capsule and fluid, daughter cyst fluid, detached germinal membrane and surrounding soft tissue reaction were noted. Observations revealed that the capsule is best seen on T2- and PD-weighted images. The daughter cysts are best demonstrated on T1-weighted images, whereas the difference in the signal intensities of parent cyst and daughter cyst fluid is better demonstrated on T2 weighted and T1-weighted images, and the detached germinal membrane is best seen on T2- and PD-weighted images. The signal characteristics of hydatid cyst morphology can help distinguish it from other entities. PMID- 11380356 TI - Interpreting urgent brain CT scans: Does review by a radiology trainee make a difference in accuracy? AB - An urgent brain CT scan is now commonly performed on patients presenting to hospital emergency departments for a wide variety of indications. At most institutions in Australia, such scans are reviewed immediately by an on-call radiologist, who is usually an accredited registrar. The value of the trainee radiologist in such a setting is unclear. In the present study, the rate of abnormal findings in a random sample of 100 brain CT scans performed on hospital patients is reviewed and the accuracy of detection of potentially urgent lesions is compared between three junior clinicians, an accredited radiology registrar and a junior radiographer, using the final radiological report as the standard of reference. At least one potentially urgent abnormality in 25% of the patients scanned was found. The RANZCR trainee recorded a significantly higher sensitivity compared to the other readers. It is concluded that an urgent brain CT is of greater value as a screening test if a contemporaneous radiological review is made available, and the implications this may have on current imaging practices are briefly considered. PMID- 11380357 TI - Computed tomography pulmonary angiography with pelvic venography in the evaluation of thrombo-embolic disease. AB - A prospective study was performed to evaluate the usefulness of CT pelvic venography (CTV) in the detection of pelvic vein thrombosis in patients referred for CT pulmonary angiography (CTPA) for suspected pulmonary embolism. Fifty consecutive patients referred for CTPA had CTV performed at the time of CTPA. All patients had duplex ultrasound (DUS) of the lower limb veins for evaluation of deep venous thrombosis (DVT) within 24 h of the CT study. Twelve (24%) of the 50 patients had pulmonary embolism diagnosed on CTPA. Associated DVT was detected in six of these patients; two cases were detected by CTV alone, while one case was detected by both CTV and DUS. The remaining three cases had DVT diagnosed by DUS alone. In the 38 patients with a negative CTPA, three patients had venous thrombus diagnosed by CTV. Of these three patients, two had a negative DUS study. CTV therefore led to a definitive imaging diagnosis of thrombo-embolic disease in two (4%) more patients. CTV adds little time and cost to the CTPA examination and leads to a moderate increase in definite imaging diagnosis of thrombo-embolic disease. PMID- 11380358 TI - Radiation oncology in Australia: workforce, workloads and equipment 1986-1999. AB - Regular national surveys of all public and private radiation oncology facilities in Australia have been carried out between 1986 and 1999. Workforce data recorded were numbers of radiation oncologists and trainees, radiation therapists, medical physicists and physics technicians, nursing staff, data managers, social workers and clerical staff. Workloads included treatments with megavoltage beams (linear accelerators, cobalt-60), orthovoltage/superficial X-rays, brachytherapy, total body irradiation and stereotactic radiosurgery. Major equipment recorded included numbers of megavoltage and orthovoltage/superficial X-ray machines, planning simulators, computerized dosimetry systems and brachytherapy equipment. The use of radiotherapy beds and the public-private mix of treatments were also documented. Data were assembled for Australia based on each individual state. Within Australia the number of public and private treatment facilities has increased by 44% from 18 in 1986 to 26 in 1999. The population has increased by 16.4%, cancer incidence by 51.8% and megavoltage workloads (fields) by 102%. The number of radiation therapists and physicists and the number of linear accelerators have, in general, increased with the growth in workloads. The number of radiation oncologists has increased by 60% from 4.5 full-time equivalent (FTE) radiation oncologists per million population in 1986 to 7.2 per million in 1999. There is currently a deficit of at least 40 radiation oncologists to be able to treat the 50% of newly diagnosed cancer patients requiring radiotherapy. In addition, a significant deficiency exists in numbers of radiation therapists, nursing staff, data managers, social workers and clerical staff. Clearly the demands for medical physicists has increased but the data are insufficient to comment on deficiencies. Despite the increases in workloads the proportion of patients with cancer receiving radiotherapy remains below 40%. A positive correlation has been shown between the proportion of newly diagnosed cancer patients treated and the number of FTE radiation oncologists, the number of megavoltage machines and number of radiation therapists. This was shown for Australia as a whole, for each state and for the years 1986 to 1999. This was also the case when total megavoltage fields was used as the dependent variable. Multiple regression analysis using the same independent variables confirmed these positive correlations. It is concluded that the low treatment rate with radiation oncology for cancer patients in Australia is due mainly to the lack of resource allocation. The stated commitment of governments and health departments to a 50% treatment rate can only become a reality if there is a concerted effort to increase the numbers of radiation oncologists, radiation therapists, megavoltage machines and support staff. Otherwise at least one in every 10 newly diagnosed cancer patients will continue to be denied adequate and equitable access to radiotherapy - in 1999 that total figure was 9400 persons. PMID- 11380359 TI - Breast radiotherapy: an Australasian survey of current treatment techniques. AB - Prior to the dissemination of evidence-based quality assurance guidelines, the Australian National Breast Cancer Centre Radiation Oncology Group conducted a process survey of breast radiotherapy treatment delivery throughout Australia. A process survey was conducted in August/September 1998. This survey comprised questions enquiring about treatment positioning, immobilization devices used, planning strategies, simulation and dose computation methods, treatment prescribing and quality assurance. The survey was sent to 123 Australian fellows of the Royal Australian and New Zealand College of Radiologists (RANZCR) and to the six directors of New Zealand radiation oncology departments. Fifty-eight questionnaires were returned of which 38 were received from individuals and 20 represented a reply from a department with a routine breast radiotherapy protocol (representing an average of 4.5 radiation oncologists per reply). The study identified great consistency between departments with respect to dose and fractionation for breast tangents. The study also identified some areas of treatment planning and delivery that varied between individuals or departments. These mainly reflected a lack of evidence in some areas of radiotherapy treatment delivery. The circulation of quality assurance guidelines will perhaps improve consistency of radiotherapy techniques in which studies have identified that technique changes improve outcome. This study identified that these areas include the taking of simulation and port films and the use of off-axis dosimetry. Further studies are required for areas of radiotherapy treatment delivery that have little evidence for or against their implementation. PMID- 11380360 TI - Carcinoid tumour of the orbital muscles: A rare occurrence. AB - Malignant carcinoid tumours are rare and usually follow an indolent course. Metastases from these tumours are uncommon, and usually involve bone, liver and lymph nodes. Orbital metastases have rarely been described, and generally involve the choroid rather than orbital structures. Our knowledge of the response of these tumours to radiotherapy is extremely limited. Two cases of orbital metastases treated with radiotherapy are presented here and the literature on the efficacy of palliative radiotherapy for this tumour is reviewed. Recommendations are made regarding an effective radiotherapy regimen for treating orbital metastases from malignant carcinoid tumours. PMID- 11380361 TI - System validation and work practice efficiency gains of a new localization method for stereotactic radiotherapy. AB - The increased procedural demands of stereotactic localization techniques when compared with conventional treatment practices reduces machine efficiency, an outcome likely to be greatly magnified by the introduction of fractionation to stereotactic techniques. Currently in Australia and New Zealand there are no guidelines for the definition of efficiency. We sought to devise a system to simultaneously validate the accuracy and efficiency of the technique. The frameless relocation methods employed in the Medtronic Sofamor Danek (MSD) stereotactic radiotherapy (SRT) system were studied in the clinical setting. Accuracy has been determined according to the accumulation of errors throughout the planning and treatment process. The clinical demands of the system (staffing and resources) were analysed relative to conventional treatment approaches. Timing studies indicate a mean time of 19.7 min for treatment of a daily SRT fraction (4-5 arcs, single isocentre). Cost and staffing requirements are similar to those for conventional radiotherapy. It is concluded that with the system used, SRT is efficient for routine clinical implementation, with the level of efficiency increasing with increasing patient numbers. It is recommended that a common acceptance standard be developed to allow cross-institutional comparison of the clinical efficiency of new treatment techniques. PMID- 11380362 TI - Quality assurance in a Radiation Oncology Unit: the Chart Round experience. AB - Quality assurance ensures that planned treatments eventuate. Programmes must include feedback loops to promptly correct any shortfall in predetermined standards. In March 1999, a weekly Chart Round was introduced to verify that certain items relevant to quality care were being completed for patients of the Head and Neck Radiotherapy Unit at the Peter MacCallum Cancer Institute. The experience was reviewed after 1 year and it was found that the initiation of Chart Rounds has assisted in raising the level of item completion from 80% to 99% in similar groups of patients treated before and after the initiation of the Chart Round. Initiation of the Chart Round has also provided a useful forum for in-house peer-review, education and effective real-time communication between medical and allied health personnel, all of which has further added to the quality of patient care. PMID- 11380363 TI - Squamous cell carcinoma of the lip. Patterns of relapse and outcome: Reporting the Westmead Hospital experience, 1980-1997. AB - Australia has one of the highest rates of squamous cell carcinoma of the lip in the world. Despite a high cure rate, many studies report relapse rates of between 5% and 20% with an associated mortality of 5-10%. The aim of this study was to review the patterns of relapse and outcome for patients treated at Westmead Hospital, Sydney. Ninety-three eligible patients were identified in a retrospective review of all lip cancer patients referred to Westmead Hospital between 1980 and 1997. Relevant data were extracted from the treatment files and included contact with referring doctors and utilizing the Cancer Council Registry. Equal numbers of patients were treated with radiotherapy alone or surgery (+/- adjuvant radiotherapy). The majority of patients were male (78.5%) with T1N0 cancers (64.5%) of the lower lip. A minority (5.4%) had nodal disease at diagnosis. Following treatment, 31 (33.3%) patients relapsed, 11 at the primary site, 18 at the regional nodes and two at both sites simultaneously. Patients treated with a combined approach (RTx/Sx) experienced a better outcome. Overall cancer-specific survival at 5 years was 85%. These findings highlight a disturbing relapse rate and mortality for a subgroup of patients diagnosed with a more aggressive form of lip cancer. Treatment and follow up of these patients should accordingly be more aggressive. PMID- 11380364 TI - Breast conservation in BRCA1 or BRCA2 mutation carriers with early stage breast cancer. AB - The role of breast conservation therapy (limited surgery and irradiation of the breast with/without axilla) in the approximately 5% of breast cancer patients who harbour a germline mutation in BRCA1 or BRCA2, is a largely unexplored area and is seen by some as controversial. The relatively high cumulative risk of second primary cancers in such patients and concern about a possible decreased ability of mutation carriers to repair DNA damage caused by radiation has fuelled this controversy. Published studies of breast conservation therapy in carriers of a mutation in BRCA1 or BRCA2 are reviewed, with particular attention to their methodology. These studies have not demonstrated any increase in radiation sensitivity of normal tissues in mutation carriers, either in terms of increased early or late toxicity or tumourigenesis. Likewise, tumour sensitivity to radiotherapy, which might be expected based on the known functions of the BRCA1 and BRCA2 genes, has not been documented to date in mutation carriers. Further, methodologically rigorous studies of large numbers of breast cancer patients who carry a mutation in BRCA1 or BRCA2 are required to fully elucidate these issues. PMID- 11380365 TI - Dose perturbation by air cavities in megavoltage photon beams: implications for cavity surface doses. AB - Gas-filled cavities in the body are known to perturb megavoltage photon beams in radiation therapy. The bowel represents one such cavity where cavity dimensions can vary throughout a treatment course. This has implications for doses to the surface of the bowel. Doses to the first 0.1 mm and 1.0 mm of cavity surfaces were calculated for 6 MV and 18 MV photon beams for a range of cavity sizes. It is found that relatively minor variations in cavity surface doses result over the typical range of rectal cavity size (2-3 cm). However, the variations in surface dose should be considered whenever bowel reactions must be considered, or when the target cells of interest line the bowel itself. PMID- 11380366 TI - Primary intraosseous meningioma of orbit and anterior cranial fossa: a case report and literature review. AB - Primary intraosseous meningiomas of the skull base are rare. Extensive involvement of the skull base by the tumour may result rarely in loss of vision. Surgical management requires extensive decompression and reconstruction. A case of primary intraosseous meningioma of orbits and anterior cranial fossa presenting with painless proptosis and loss of vision is presented here and the literature reviewed. PMID- 11380367 TI - Fatal cerebral air embolism as a complication of transbronchoscopic lung biopsy: a case report. AB - A rare complication of transbronchoscopic lung biopsy, namely a cerebral air embolism, is presented. The course of events following the embolic episode in the form of a fall in blood pressure, bradycardia and convulsions is documented, as is the presence of an air emboli on the CT scan of the brain with subsequent resorption of the emboli on the follow-up scan. The salient features of the case are the rarity of the complication and the excellent temporal depiction of imaging findings on CT scan demonstrated as resorption of air emboli and subsequent watershed territory infarcts. PMID- 11380368 TI - Percutaneous ultrasound-guided thrombin injection for coagulation of post traumatic pseudoaneurysms. AB - Most peripheral pseudoaneurysms are iatrogenic or, less commonly, post-traumatic. Pseudoaneurysms are associated with characteristic findings of a pulsatile palpable mass and an audible to and fro murmur. The diagnosis can be easily confirmed using colour duplex ultrasound. A successful treatment for the coagulation of pseudoaneurysms as an alternative to ultrasound-guided manual compression is described. The method involves ultrasound-guided direct percutaneous injection of a small quantity of thrombin through a fine needle. This technique is simple and effective, resulting in rapid occlusion of the pseudoaneurysm. Two cases are reported. PMID- 11380369 TI - Sonographic diagnosis of diastematomyelia in utero: a case report and literature review. AB - Fetal diastematomyelia is a rare form of spinal dysraphism that is characterized by a complete or incomplete division of the spinal cord by an osseous or fibrocartilaginous septum. A case of diastematomyelia, which was detected on the routine third trimester detailed ultrasound scan, is presented. The diagnosis was based on the detection of an echogenic focus in the posterior aspect of the spine in association with widening of the interpedicular vertebral space. The case illustrates that diastematomyelia can occur in the absence of overt spina bifida and that prenatal detection will allow timely postnatal investigation and treatment. Prenatal literature is further reviewed to assess the clinical significance of this finding. PMID- 11380370 TI - Pseudo-Kaposi's sarcoma: the association of arterio-venous malformations with skin lesions resembling Kaposi's sarcoma. AB - A patient is described with skin lesions resembling Kaposi's sarcoma (KS). Arteriography revealed multiple arteriovenous malformations in the affected limb. This condition has been termed pseudo-Kaposi's sarcoma. Although reports of this condition have appeared in dermatological journals, as yet we are unaware of any account in the radiological literature. This paper presents a case and reviews the literature. In suspected cases of KS in which the history or clinical features are atypical, this unusual condition should be considered and a search made for the diagnostic vascular lesions. PMID- 11380371 TI - Multifocal malignant extra-adrenal paragangliomas of the Organ of Zuckerkandl and urinary bladder. AB - A case of multifocal malignant extra-adrenal paragangliomas involving the Organ of Zuckerkandl and urinary bladder is presented. The role and order of imaging investigations of functioning extra-adrenal paraganglioma is discussed. PMID- 11380372 TI - Paradoxical embolism: computed tomography demonstration. AB - Paradoxical emboli are rare and often presumptively diagnosed. A case of paradoxical embolism, in which both the arterial and venous emboli were documented on CT, is described. PMID- 11380373 TI - Arachnoiditis associated with arachnoid cyst formation and cord tethering following myelography: magnetic resonance features. AB - The reported incidence of clinically significant arachnoiditis following oil based contrast media myelography is approximately 1%. The incidence of arachnoiditis in the dorsal spine is even more rare. Acquired arachnoid cyst formation can occur with arachnoiditis of various aetiologies. A case is presented of a patient with chronic backache, radiculopathy, weakness and muscle wasting, who had undergone myelography with oil-based contrast media about 28 years ago. There was no history of spinal surgery, spinal canal stenosis, disc disease or trauma, all of which are known to increase the risk of arachnoiditis. Magnetic resonance imaging features are presented in this case of arachnoiditis with arachnoid cyst formation, and cord tethering. PMID- 11380374 TI - Malignant chondroid syringoma of the skin: magnetic resonance imaging features. AB - Skin tumours are usually divided into melanoma and non-melanoma types. Malignancies of the adnexal structures, of which sweat gland tumours are an example, are characterized under the non-melanoma types. Sweat gland malignancies are rare tumours that are usually associated with a poor prognosis. Given the rarity of these tumours, MRI findings of such tumours have not been described previously in the literature. We present a case report of an unusual malignant tumour of sweat gland origin known as a malignant chondroid syringoma of the skin with described MRI features. The MRI features are non-specific depicting intermediate signal intensity, changes on the proton density sequence and increased signal on the T2 and STIR-weighted sequences. Although these imaging features are characteristic of most soft tissue masses MRI can, in most cases, accurately depict the anatomic extent and identify tissue of origin, depth of invasion and relation to adjacent structures, such as muscles and bones. Thus high-resolution MRI of the skin in the future can be extremely helpful in characterizing and staging dermal neoplasms. PMID- 11380375 TI - Pituitary tuberculoma mimicking adenoma: magnetic resonance imaging. AB - A case of a pituitary mass with clinical and MRI findings consistent with adenoma is presented. Transnasal transphenoidal excision biopsy showed it to be a pituitary tuberculoma. The patient was treated with antituberculous drugs, and a follow-up MRI 18 months later showed good response. MRI features and a review of published reports of the sellar tuberculoma are briefly discussed. PMID- 11380376 TI - Cavernous haemangiomas (angiomas) of the brain. AB - This paper highlights the importance of cavernous haemangiomas as clinically significant lesions and the role of imaging, particularly MRI, in suggesting the diagnosis. An understanding of the pathology of these lesions helps to explain the features demonstrated by imaging techniques. PMID- 11380377 TI - Violating "the Code": breastfeeding, ethics, and choices. PMID- 11380378 TI - Assessing effective care in normal labor: the Bologna score. AB - The intention of the "Bologna score" is to quantify, both in an individual labor and in a wider population, the extent to which labors have been managed as if they are normal as opposed to complicated. In this way it may be possible to assess both attitudes and practices within a maternity service toward the effective care of normal labor. A scoring system for normal labor was proposed at the World Health Organization (Regional Office for Europe) Task Force Meeting on Monitoring and Evaluation of Perinatal Care, held in Bologna in January 2000. This paper describes conceptual development of the scale. Recommendations for future evaluation of the Bologna score's validity and potential include field testing globally, comparison with the Apgar score, and evaluation of the relative weight contributed by each of the five measures comprising the Bologna score. PMID- 11380379 TI - Immersion in water in the first stage of labor: a randomized controlled trial. AB - BACKGROUND: Current forms of analgesia often have significant side effects for women in labor. Bathing in warm water during labor has been reported to increase a woman's comfort level and cause a reduction in painful contractions. The objective of this trial was to compare immersion in warm water during labor with traditional pain management for a range of clinical and psychological outcomes. METHODS: A prospective randomized controlled trial of 274 pregnant women, who were free from medical and obstetric complications and expecting a singleton pregnancy at term, was conducted at the Women's and Children's Hospital, a maternity tertiary referral center in Adelaide, South Australia. Women in labor were randomized to an experimental group who received immersion in a bath or to a nonbath group who received routine care. Pharmacological pain relief was the primary outcome that was measured, and secondary outcomes included maternal and neonatal clinical outcomes, factors relating to maternal and neonatal infectious morbidity, psychological outcomes, and satisfaction with care. RESULTS: The use of pharmacological analgesia was similar for both the experimental and control groups; 85 and 77 percent, respectively, used major analgesia. No statistical differences were observed in the proportion of women requiring induction and augmentation of labor or in rates of perineal trauma, length of labor, mode of delivery, or frequency of cardiotocographic trace abnormalities. Neonatal outcomes (birthweight, Apgar score, nursery care, meconium-stained liquor, cord pH estimations) revealed no statistically significant differences. Infants of bath group women required significantly more resuscitation than routine group women. Routine group women rated their overall experience of childbirth more positively than bath group women. Psychological outcomes, such as satisfaction with care or postnatal distress, were the same for both groups. CONCLUSION: Bathing in labor confers no clear benefits for the laboring woman but may contribute to adverse effects in the neonate. PMID- 11380380 TI - Maternity care practices: implications for breastfeeding. AB - BACKGROUND: Many United States mothers never breastfeed their infants or do so for very short periods. The Baby-Friendly Hospital Initiative was developed to help make breastfeeding the norm in birthing environments, and consists of specific recommendations for maternity care practices. The objective of the current study was to assess the impact of the type and number of Baby-Friendly practices experienced on breastfeeding. METHODS: A longitudinal mail survey (1993 1994) was administered to women prenatally through 12 months postpartum. The study focused on the 1085 women with prenatal intentions to breastfeed for more than 2 months who initiated breastfeeding, using data from the prenatal and neonatal periods. Predictor variables included indicators of the absence of specific Baby-Friendly practices (late breastfeeding initiation, introduction of supplements, no rooming-in, not breastfeeding on demand, use of pacifiers), and number of Baby-Friendly practices experienced. The main outcome measure was breastfeeding termination before 6 weeks. RESULTS: Only 7 percent of mothers experienced all five Baby-Friendly practices. The strongest risk factors for early breastfeeding termination were late breastfeeding initiation and supplementing the infant. Compared with mothers experiencing all five Baby Friendly practices, mothers experiencing none were approximately eight times more likely to stop breastfeeding early. Additional practices decreased the risk for early termination. CONCLUSION: Increased Baby-Friendly Hospital Initiative practices improve the chances of breastfeeding beyond 6 weeks. The need to work with hospitals to increase adoption of these practices is illustrated by the small proportion of mothers who experienced all five practices measured in this study. PMID- 11380381 TI - Women's preference for a cesarean section: incidence and associated factors. AB - BACKGROUND: Few studies have examined women's preferences for birth. The object of this study was to determine the incidence of women's preferred type of birth, and the reasons and factors associated with their preference. METHODS: Three hundred and ten women between 36 and 40 weeks' gestation were recruited from the antenatal clinic of a major metropolitan teaching hospital and the consulting rooms of six private obstetricians in Brisbane, Australia. Participants completed a questionnaire asking about their preferred type of birth, reasons for their preference, preparation for childbirth, level of anxiety and concerns, and the influence of the primary caregiver. RESULTS: Two hundred and ninety women (93.5%) preferred a spontaneous vaginal birth; 20 women (6.4%) preferred a cesarean section. Of the latter group, most had a current obstetric complication or experienced a previously complicated delivery (p <0.001); 1 woman (0.3%) preferred a cesarean section in the absence of any known current or previous obstetric complication. Women who preferred a cesarean section were more anxious, were generally poorly informed of the risks of this procedure, and/or overestimated the safety of the procedure. CONCLUSIONS: Women who preferred a cesarean section were more likely to have experienced this type of birth previously and to have negative feelings about it. To decrease women's preference for a cesarean section, practitioners should reduce the primary cesarean delivery rate and improve the quality of emotional care for women who require a cesarean section. Caregivers should engage in a sensitive discussion of the risks and benefits of various birth options, including a vaginal birth after cesarean, with women who have previously experienced a cesarean birth before they make decisions about mode of delivery in a subsequent pregnancy. PMID- 11380382 TI - Do women get posttraumatic stress disorder as a result of childbirth? A prospective study of incidence. AB - BACKGROUND: Recent research suggests that a proportion of women may develop posttraumatic stress disorder after birth. Research has not yet addressed the possibility that postpartum symptoms could be a continuation of the disorder in pregnancy. This study aimed to test the idea that some women develop posttraumatic stress disorder as a result of childbirth, and to provide an estimate of the incidence using a prospective design, which controls for the disorder in pregnancy. METHOD: This prospective study assessed 289 women at three time points: 36 weeks gestation and 6 weeks and 6 months postpartum. The prevalence of posttraumatic stress disorder was assessed by questionnaire at each time point, and the incidence was examined after removing women who had severe symptoms of posttraumatic stress disorder or clinical depression in pregnancy. RESULTS: After removing women at the first time point, 2.8 percent of women fulfilled criteria for the disorder at 6 weeks postpartum and this decreased to 1.5 percent at 6 months postpartum. CONCLUSIONS: The results suggest that at least 1.5 percent of women may develop chronic posttraumatic stress disorder as a result of childbirth. It is important to increase awareness about the disorder and to give health professionals access to simple screening tools. Intervention is possible at several levels, but further research is needed to guide this intervention. PMID- 11380383 TI - Eating and drinking in labor: the influence of caregiver advice on women's behavior. AB - BACKGROUND: Although there is much debate about eating and drinking during labor, little scientific data about its influence on the course of labor exist. In The Netherlands, most midwives and obstetricians allow women to eat and drink during normal labor. The objective of this study was to examine whether or not women were actively advised to eat and drink and if this advice affected eating and drinking behavior. METHODS: A randomly selected group of midwives and obstetricians from across The Netherlands identified 211 consecutive nulliparous women to participate in the study. In a questionnaire with open-ended questions, women were asked after their delivery whether or not they were advised about eating and drinking during labor, and if so, about the nature of this advice and what they had consumed. Data were analyzed at the Leyenburg Hospital in The Hague. RESULTS: Sixty-six percent of the women were not given advice about eating and drinking during labor. Women who were given advice usually followed it. In the total group, 37 percent of the women had intake other than water and of these, 75 percent ate solid food. After adjusting for other prognostic factors, the incidence of an instrumental delivery due to a nonprogressing second stage was lower in women with caloric intake (13% vs 24%, p = 0.04). CONCLUSION: The study design did not enable us to draw conclusions about the cause and effect between caloric intake and labor progress. Scientific data with respect to the giving of evidence-based advice about eating and drinking during labor are lacking. Should such advice become available, women are likely to follow it. PMID- 11380385 TI - Flat tires and other deflations. PMID- 11380384 TI - A waste of life: fathers' experience of losing a child before birth. AB - BACKGROUND: The prenatal loss of an expected child entails parental despair and grief. The grief after a stillborn child is sometimes described as a "forgotten form of grief" and the fathers as the "forgotten mourners." Our aim was to describe how fathers experienced losing a child as a result of intrauterine death. METHOD: Eleven men were interviewed 5 to 27 months after the intrauterine death of their child during weeks 32 to 42 of pregnancy. The interviews were analyzed using a phenomenological methodology. RESULTS: After being informed of the infant's death, most fathers first wanted their partners to have a cesarean section, but all later thought that it would be right for the child to be delivered vaginally. A strong feeling of frustration and helplessness came over them during and after the delivery. Several men found meaning and relief in their grief by supporting their partner. Tokens of remembrance from the child were invaluable, and fathers appreciated that the staff collected these items, even if the parents declined them. The perceived prerequisite for resuming their everyday lives consisted of the support they received from the hospital staff and precious memories of the child. The most important comfort in their grief was a good relationship with their partner. Some fathers missed having a man to talk to both at the time of the stillbirth and subsequently. CONCLUSION: The fathers' general trust in life and the natural order was suddenly and unexpectedly severely tested by the death of their child, which they perceived as a terrible waste of life. They sought understanding as grieving men and fathers from both the hospital personnel and their partners, as well as from relatives. Being able to protect their partner and to grieve in their own way was important to the fathers. PMID- 11380386 TI - Consumer-professional partnership to improve research: the experience of the Cochrane Collaboration's Pregnancy and Childbirth Group. PMID- 11380387 TI - Osteoclasts in Paget's disease of bone. PMID- 11380388 TI - Pure red cell aplasia associated with small lymphocytic lymphoma. PMID- 11380389 TI - Woolf, the adversarial system and the concept of blame. PMID- 11380390 TI - CD40 ligand (CD154) and tumour necrosis factor-related apoptosis inducing ligand (Apo-2L) in haematological malignancies. PMID- 11380391 TI - Alk+ CD30+ lymphomas: a distinct molecular genetic subtype of non-Hodgkin's lymphoma. PMID- 11380392 TI - Increased bone marrow vascularization in patients with acute myeloid leukaemia: a possible role for vascular endothelial growth factor. AB - The present study demonstrated that the vessel number in bone marrow biopsies from acute myeloid leukaemia (AML) patients (n = 23) was significantly increased at diagnosis compared with normal bone marrow (P = 0.019) and was restored to normal levels after achieving complete remission (P = 0.03). The in vitro angiogenic potential of culture supernatant of AML cells was assessed using endothelial cell (EC) migration and proliferation assays. Increased EC migration and EC proliferation was induced in 7/20 and 19/20 AML supernatents respectively. The degree of in vivo neovascularization did not correlate with the ability of AML cells to stimulate in vitro endothelial cell migration and/or proliferation. This might be in part a result of the heterogeneous pattern of angiogenic factors produced by AML cells. The expression of different angiogenic factors was studied using reverse transcription polymerase chain reaction. Cells from 17/20 AML patients showed wide variation in spontaneous vascular endothelial growth factor (VEGF) expression, 4/19 expressed varied spontaneous blastic fibroblast growth factor mRNA levels and all patient samples showed spontaneous interleukin 8 mRNA expression. All AML samples expressed matrix metalloproteinase (MMP)-2 and/or MMP 9. VEGF mRNA expression correlated well with protein level (P = 0.006). A correlation was found between the degree of VEGF expression and neoangiogenesis (correlation coefficient = 0.448, P = 0.05). These results suggest that malignant cell proliferation, angiogenesis and VEGF expression are linked in AML and might contribute to the growth advantage of the malignant counterpart as a result of the paracrine production of growth factors produced by the surrounding endothelial cells. PMID- 11380393 TI - Spectral karyotyping and fluorescence in situ hybridization detect novel chromosomal aberrations, a recurring involvement of chromosome 21 and amplification of the MYC oncogene in acute myeloid leukaemia M2. AB - Recurring chromosomal aberrations are of aetiological, diagnostic, prognostic and therapeutic importance in acute myeloid leukaemia (AML). However, aberrations are detected in only two thirds of AML cases at diagnosis and recurrent balanced translocations in only 50%. Spectral karyotyping (SKY) enables simultaneous visualization of all human chromosomes in different colours, facilitating the comprehensive evaluation of chromosomal abnormalities. Therefore, SKY was used to characterize 37 cases of newly diagnosed AML-M2, previously analysed using G banding. In 15/23 patients it was possible to obtain metaphases from viably frozen cells; in 22 additional cases, fixed-cell suspensions were used. Of the 70 chromosomal aberrations identified by SKY, 30 aberrations were detected for the first time, 18 aberrations were redefined and 22 were confirmed. SKY detected two reciprocal translocations, t(X;3) and t(11;19). In five cases, eight structural aberrations resulted in partial gains of chromosome 21, six of which were undetected by G-banding. In 4/5 cases, these resulted in copy number increases for AML1. Amplification of MYC was detected in three cases. Using SKY and FISH, clonal aberrations were identified in 5/18 cases with a presumed normal karyotype; 3/5 aberrations were of known unfavourable prognostic significance. Karyotypes were entered into a custom-designed SKY database, which will be integrated with other cytogenetic and genomic databases. PMID- 11380394 TI - Unusual T-cell receptor-delta gene rearrangement patterns revealed by screening of a large series of childhood acute lymphoblastic leukaemia by multiplex polymerase chain reaction. AB - Rearrangements of the T-cell receptor (TCR) and immunoglobulin genes are considered as useful clonal markers in lymphoproliferative disorders of B- and T cell lineage, and are frequently used for the detection of minimal residual disease (MRD). In this paper, we report on the unexpected results of an extensive analysis of TCR-delta chain gene rearrangement frequencies and patterns in leukaemic bone marrow DNA samples collected from 438 children with initial (n = 112) or relapsed (n = 326) acute lymphoblastic leukaemia (ALL). By applying a previously described multiplex polymerase chain reaction, the overall incidence of non-deleted TCR-delta gene rearrangements in ALL was 47% (206/438), 52% in initial ALL (58/112) and 45% in relapsed ALL (148/326). As expected, the majority of B-cell precursor (BCP) ALL had incomplete Vdelta2-Ddelta3 or Ddelta2-Ddelta3 TCR-delta gene rearrangements, whereas most T-ALL showed complete rearrangements of the TCR-delta gene locus (Vdelta1-Jdelta1, Vdelta2-Jdelta1, Vdelta3-Jdelta1). However, unexpectedly, 5/206 rearranged TCR-delta alleles in BCP-ALL showed a complete Vdelta-(Ddelta)-Jdelta gene rearrangement pattern, and 3/31 T-ALL had an incomplete recombination. Theoretically, complete TCR-delta gene rearrangements should not occur in cells other than T-lymphocytes and have only been reported once previously in BCP-ALL. The data contribute to the discussion about the reliable screening for clonal markers in ALL. PMID- 11380395 TI - INK4A/ARF deletions are acquired at relapse in childhood acute lymphoblastic leukaemia: a paired study on 25 patients using real-time polymerase chain reaction. AB - Current risk-adjusted intensive therapies for childhood acute lymphoblastic leukaemia (ALL) are expected to result in an event-free survival of greater than 75%. In sharp contrast, relapsed paediatric ALL is a difficult disease to treat. In this study, 25 paediatric patients with ALL were analysed at diagnosis and relapse for their p16 (exon 2) status using the most accurate method of detection, real-time polymerase chain reaction (PCR). The median time to relapse for the group was 27 months. At diagnosis, the incidence of p16 homozygous and hemizygous deletion in this group was 32% and 20% respectively. The incidence of homozygous p16 deletion at relapse was 64%. A large number of patients, eight of 16 (50%), developed p16 homozygous deletion at relapse. Of those eight patients, four were hemizygous and four were germline at diagnosis. At diagnosis, those patients with a homozygous or hemizygous p16 deletion relapsed sooner than those germline for p16. We have shown that p16 alterations are frequently present in relapsed lymphoblastic leukaemia in children. PMID- 11380396 TI - Hypermethylation of the calcitonin gene in acute lymphoblastic leukaemia is associated with unfavourable clinical outcome. AB - We analysed calcitonin (CALC1) gene hypermethylation using semiquantitative differential polymerase chain reaction in 105 patients with adult (n = 49) and childhood (n = 56) acute lymphoblastic leukaemia (ALL), and studied the association of CALC1 hypermethylation with clinical presentation features and disease outcome. We also investigated the possible relationship between CALC1 methylation status and expression of the cell cycle inhibitor gene p57KIP2. We observed CALC1 hypermethylation in bone marrow cells from 43% (45 out of 105) of ALL patients. Clinical, molecular and laboratory features did not differ significantly between hypermethylated and hypomethylated patients, only T-cell lineage was associated with hypermethylation (14% vs. 47%, P = 0025). Complete remission rate was similar in both groups although hypermethylated patients had a higher relapse rate (68% vs. 19%, P < 0.00001) and mortality rate (55% vs. 36%, P = 0.06) than hypomethylated patients. Estimated disease-free survival (DFS) at 6 years was 66.1% for hypomethylated patients and 5.3% for hypermethylated patients (P < 0,00001). Multivariate analysis from potential prognostic factors demonstrated that CALC1 methylation status was an independent prognostic factor in predicting DFS (P = 0.0001). Separate analysis of adult and childhood ALL patients showed similar results to the whole series. In addition, hypermethylated patients showed downregulation of p57KIP2 expression. Our results suggest that CALC1 gene hypermethylation is associated with an enhanced risk of relapse independently of known poor-prognostic factors and we describe, for the first time, a possible implication of the p57KIP2 gene in the genesis and prognosis of ALL. PMID- 11380397 TI - Pharmacological basis for cladribine resistance in a human acute T lymphoblastic leukaemia cell line selected for resistance to etoposide. AB - Cross-resistance between different classes of anti-neoplastic agents can jeopardize successful combination cancer chemotherapy. In this study, we observed an unexpected cross-resistance between the podophyllotoxine derivative etoposide (VP) and the nucleoside analogue cladribine (CdA) in CCRF-CEM cells developed for resistance to VP. The resistant cells also displayed 14- and twofold resistance to cytarabine (ara-C) and gemcitabine respectively. Closer analysis of these cells showed that they contained lower amounts of topoisomerase (topo) IIalpha (P < 0.001) and beta protein (P < 0.026), formed substantially lower amounts of the topo II-DNA complex, and had a markedly decreased level of Fas (CD95/APO-1) ligand mRNA expression. Interestingly, Fas expression in the resistant cells did not differ from that in the parental cell line. No differences were observed in the accumulation/efflux of daunorubicin or in the gene expressions of P glycoprotein, multidrug resistance-associated protein and the lung resistance related protein. The activity of deoxycytidine kinase (dCK), responsible for activation of CdA and ara-C, was the same for resistant and wild-type cells. However, there was an increase in the activity of the cytosolic 5'-nucleotidases (5'-NT), responsible for deactivation of nucleotides, amounting to 206% (P < 0.001) for the high Km and 134% (P < 0.331) for the low Km 5'-NT in resistant cells. The high Km 5'-NT is probably responsible for the decreased amount of the active metabolite CdA 5'-triphosphate [40% decreased (P < 0.045)], as well as for other purine ribonucleosides and deoxyribonucleosides triphosphates in the resistant cells. In contrast, a significantly higher deoxycytidine triphosphate (dCTP) level (167%, P < 0.001) was observed in the resistant cells. Thus, this study suggests that the major cause of resistance to the nucleoside analogues CdA and ara-C in cells selected for resistance to VP is a result of metabolic alterations producing increased activity of 5'-NT and higher dCTP levels. Furthermore, these results indicate that there is a common factor in the regulation of nucleotide-degrading enzymes and DNA topoisomerases, which may be altered in cross-resistant cells. PMID- 11380398 TI - The prognostic impact of karyotypic subgroups in myelodysplastic syndromes is strongly modified by sex. AB - The prognostic impact of karyotypic patterns in a consecutive series of 389 adult myelodysplastic syndromes (MDS) was investigated. Time period did not significantly influence the survival times. In the analyses, the MDS cases were subdivided into the cytogenetic subgroups used in the International Prognostic Scoring System, i.e. favourable [-Y, del(5q) or del(20q) as single aberrations or normal karyotype, n = 241], poor [-7, del(7q), der(1;7) or complex karyotypes, i.e. > or = three abnormalities, n = 89] and intermediate (other aberrations, n = 59). The survival times correlated well with the prognostic subgroups, confirming that the cytogenetic classification was valid. Expressed as hazard ratios (HRs), with the favourable subgroup as the reference, the intermediate and poor subgroup HRs increased to 1.5 (95% confidence interval, 1.1-2.1) and 3.2 (2.4-4.1) respectively. Sex, age, morphological subtype and smoking habits significantly modified this prognostic impact. Shorter survival was detected for men in the favourable and the intermediate subgroups, but not in the poor prognosis subgroup. Using women in the favourable subgroup as the reference and adjusting for age, the HR for men was 1.6 (1.2-2.1) in the favourable subgroup. Adjusting for smoking habits as well decreased the HR to 1.4 (1.1-2.0) and, when also excluding cases with del(5q) as the sole anomaly, no significant difference could be discerned [HR 1.2 (0.9-1.6], suggesting that the better outcome for women in the favourable subgroup was mainly as a result of the '5q-syndrome' and to smoking habits. In the intermediate subgroup, the corresponding HRs were 3.0 (1.5 6.0) when adjusted for age and 2.7 (1.3-5.5) when also adjusted for smoking habits. Different survival times between men and women have never previously been reported for this MDS group. Although it remains to be elucidated whether environmental and/or constitutional factors cause the observed sex-related difference, these observations have obvious clinical ramifications, not least in designing and evaluating therapy protocols. PMID- 11380399 TI - Mutation analysis of C-KIT in patients with myelodysplastic syndromes without mastocytosis and cases of systemic mastocytosis. AB - The proto-oncogene C-KIT encodes a tyrosine kinase receptor that is expressed on mast cells and haematopoietic stem cells and can show somatic mutations in patients with mastocytosis. Only scattered information is available about mutations in C-KIT in patients with other myeloid neoplasms. Moreover, the prevalence of mutations in C-KIT in bone marrow specimens of individuals with systemic mastocytosis is largely unknown. Using sequence analysis, we have screened cDNAs of the C-KIT domain encompassing codon 510-626 and codon 763-858 in bone marrow (BM) mononuclear cells (MNCs) of patients with myelodysplastic syndromes (n = 28) and patients with systemic mastocytosis (n = 12) for the presence of mutations. Furthermore, restriction fragment length polymorphism analysis was applied for identification of the C-KIT 2468A-->T and the C-KIT 1700T-->G mutation, as well as the C-KIT 1642A-->C polymorphism. All 11 patients with systemic indolent mastocytosis tested positive for C-KIT 2468A-->T. In contrast, no mutation was identified in the case of aggressive mastocytosis. Among patients with myelodysplastic syndromes, no patient showed a somatic mutation in C-KIT. The allele frequency for C-KIT 1642A-->C among the entire patient population was 0.038 and was 0.125 among age- and sex-matched healthy controls. Our data demonstrate that myelodysplastic syndromes without histological or cytological evidence of mastocytosis do not exhibit somatic mutations in exons 10, 11, 12, 16, 17 and 18 of C-KIT. In contrast, BM MNCs of patients with systemic indolent mastocytosis were all positive for C-KIT 2468A- >T and negative for additional mutations in these exons. The C-KIT 1642A-->C polymorphism is not associated with myelodysplastic syndrome or systemic mastocytosis. PMID- 11380400 TI - Fluorescence in situ hybridization analysis of 25 cases of idiopathic myelofibrosis and two cases of secondary myelofibrosis: monoallelic loss of RB1, D13S319 and D13S25 loci associated with cytogenetic deletion and translocation involving 13q14. AB - To identify a commonly deleted region of 13q14 in idiopathic myelofibrosis (IMF), we used fluorescence in situ hybridization analysis to test for deletion of the RB1 and BRCA2 genes, and the microsatellite loci D13S319 and D13S25, in a series of 25 patients. A further two patients with myelofibrosis secondary to polycythaemia vera and essential thrombocythaemia with reciprocal 13q translocations were studied in an attempt to further define the CDR. Twenty out of 21 patients with a cytogenetically normal chromosome 13 failed to show allelic loss with any of the four probes. In contrast, all four cases with cytogenetic deletion of 13q14 and both cases with 13q translocations involving 13q14 exhibited loss of RB1, D13S319 and D13S25. Loss of the BRCA2 locus was present in a single case only. Our results indicate that cryptic deletions of the 13q14 in myelofibrosis are rare. In addition, the genetic loss associated with cytogenetic 13q14 deletions or reciprocal translocations involving 13q14 is large and encompasses the gene-rich region around RB1, D13S319 and D13S25. PMID- 11380401 TI - Technetium-99m-sestamethoxyisobutylisonitrile scan as a predictor of chemotherapy response in malignant lymphomas compared with P-glycoprotein expression, multidrug resistance-related protein expression and other prognosis factors. AB - The purpose of the present study was to predict the response of malignant lymphomas (MLs) to chemotherapy using technetium-99m methoxyisobutylisonitrile (Tc-MIBI) scan and to compare it with the predictive ability of P-glycoprotein (P gp) expression, multidrug resistance-related protein (MRP) expression and other prognosis factors. Twenty-five ML patients were enrolled in this study prior to initiation of chemotherapy. Images were obtained 10 min after intravenous injection of Tc-MIBI, interpreted visually and the tumour-to-background (T/B) ratios calculated. Immunohistochemical analyses were performed on sections of the biopsy specimens to determine P-gp and MRP expression. Chemotherapy response was evaluated in the first 1-2 years after completion of chemotherapy. The mean T/B ratio of the 15 patients with a good response (3.3 +/- 0.6) was significantly higher than that of the 10 patients with a poor response (1.2 +/- 0.1). All 15 patients with a good chemotherapy response had positive Tc-MIBI scan results and negative P-gp and MRP expression. All 10 patients with a poor response had negative Tc-MIBI scan results and either positive P-gp or MRP expression. Other prognosis factors showed no significant difference in the incidence of good and poor responses. Tc-MIBI scan results represent P-gp or MRP expression more accurately than other prognosis factors and predict the chemotherapy response in ML patients. PMID- 11380403 TI - bcl-2 expression in non-Hodgkin's lymphomas is not associated with bcl-2 gene rearrangements. AB - Previous reports have associated bcl-2 gene rearrangements found in non-Hodgkin's lymphomas with an inappropriately elevated bcl-2 expression compared with the mature B-cell stage of development. This study investigates bcl-2 expression in non-Hodgkin's lymphomas (NHL) without bcl-2 gene rearrangements. Molecular analysis in 168 patients with NHL revealed 45 patients without bcl-2 gene rearrangements in which additional immunostaining for bcl-2 protein was possible. An unexpectedly high prevalence (39/45) of bcl-2 expression was found. The levels and patterns of bcl-2 expression were not specific for the histological type of NHL and were similar to those shown in comparable cases with bcl-2 gene rearrangements. In conclusion, bcl-2 expression is not specific for NHL bearing bcl-2 gene rearrangements. This finding implicates the existence of other deregulating control mechanisms of bcl-2 expression, more important than bcl-2 gene rearrangements. PMID- 11380402 TI - A new G-CSF-supported combination chemotherapy, LSG15, for adult T-cell leukaemia lymphoma: Japan Clinical Oncology Group Study 9303. AB - This phase II trial was performed to evaluate the efficacy of a new granulocyte colony-stimulating factor (G-CSF)-supported multi-agent chemotherapy protocol, LSG15, for aggressive adult T-cell leukaemia-lymphoma (ATL). Ninety-six previously untreated patients with aggressive ATL were enrolled and grouped as: acute type (58), lymphoma type (28) and unfavourable chronic type (10). Therapy consisted of seven cycles of VCAP (vincristine, cyclophosphamide, doxorubicin and prednisone), AMP (doxorubicin, ranimustine and prednisone) and VECP (vindesine, etoposide, carboplatin and prednisone). G-CSF was administered during the intervals between chemotherapy until neutrophil reconstitution was achieved. Eighty-one per cent of the 93 eligible patients responded [95% confidence interval (CI), 71.1-88.1%], with 33 patients obtaining complete response (35.5%) and 42 obtaining partial response (45.2%). The median survival time (MST) after registration was 13 months and the median follow-up duration of the 20 surviving patients was 4.2 years (range 2.8-5.6). Overall survival at 2 years was estimated to be 31.3% (95% CI, 22.0-40.5%). Grade 4 haematological toxicity of neutropenia and thrombocytopenia were observed in 65.3% and 52.6% of the patients respectively, but grade 4 non-haematological toxicity was observed in only one patient. LSG15 is feasible with mild non-haematological toxicity and improved the clinical outcome of ATL patients. MST and overall survival at 2 years were superior to those obtained by our previous trials. PMID- 11380404 TI - Reduced susceptibility to apoptosis correlates with kinetic quiescence in disease progression of chronic lymphocytic leukaemia. AB - The role of apoptosis and cell kinetics in the mechanisms of disease progression of chronic lymphocytic leukaemia (CLL) is still unclear. In the present study, we investigated the susceptibility of leukaemic cells taken from 75 CLL patients with either stable (STD) or progressive disease (PRD) to enter apoptosis. Particular attention was paid to the relationship between cell cycle status and autologous serum (AS). The susceptibility to enter apoptosis was significantly greater in STD than in PRD, both in standard medium (mean = 23.62% +/- 14.7 versus 14.23% +/- 7.2; P = 0.02) and in the presence of AS (mean = 23.03% +/- 17.9 versus 11.27% +/- 7.6; P = 0.01). Furthermore, cell kinetics studies revealed a higher quiescence in PRD than in STD cases, both in terms of a lower RNA content (P = 0.04) and of higher expression of the negative cell cycle regulator p27kip1 (P = 0.03). These kinetic differences were confirmed by short term in vitro culture both in fetal calf serum and in AS. The results of this study indicate that CLL cells from PRD cases are characterized by a higher degree quiescence and much lower susceptibility to apoptosis when compared with STD ones. In this context, AS does not appear to play a specific role. The association between these kinetic characteristics and disease progression in CLL prompts further studies to establish whether higher quiescence may be responsible for the decreased susceptibility of PRD cells to enter apoptosis. PMID- 11380405 TI - Bcl-2 expression correlates positively with serum basic fibroblast growth factor (bFGF) and negatively with cellular vascular endothelial growth factor (VEGF) in patients with chronic lymphocytic leukaemia. AB - A large proportion of B-chronic lymphocytic leukaemia (B-CLL) cells express the anti-apoptotic protein Bcl-2. Basic fibroblast growth factor (bFGF) has been shown to upregulate the expression of Bcl-2 in B-CLL cell lines. Vascular endothelial growth factor (VEGF) has been shown to enhance the survival of endothelial cells by upregulating the expression of Bcl-2. In the present study, we measured serum and cellular levels of bFGF and VEGF in 85 patients with CLL using a commercial quantitative sandwich enzyme immunoassay technique. Levels of Bcl-2 were also assayed concomitantly using Western blot analysis. The mean serum level of bFGF was 53.4 pg/ml (range 0-589) and that of VEGF 459.2 pg/ml (range 33 1793). The mean cellular level of bFGF was 158.3 pg/2 x 105 cells (range 0.8-841) and VEGF, 42.4 pg/2 x 105 cells (range 0-244). A high correlation was found between serum and cellular bFGF levels (P < 0.001), but not between the corresponding VEGF levels. Twenty-nine of 69 patients (42%) evaluated for Bcl-2 level, expressed it. The Bcl-2 level was positively correlated with the serum bFGF level (P = 0.007). However, surprisingly there was a negative correlation between Bcl-2 expression and intracellular VEGF level (P = 0.003). A positive correlation was also found between serum bFGF and disease follow-up time and log white blood cell count. These findings indicate that in CLL there is a correlation between angiogenesis-related factors and apoptosis-related protein expression, and elevated bFGF levels may account for the elevated Bcl-2 levels. PMID- 11380406 TI - Anticipation in familial chronic lymphocytic leukaemia. AB - Anticipation, a phenomenon in which an inherited disease is diagnosed at an earlier age in each successive generation of a family, has been demonstrated in certain heritable neurological disorders and in multiple myeloma, non-Hodgkin's lymphoma and other haematological neoplasms. The present study was conducted to determine whether anticipation occurs in familial chronic lymphocytic leukaemia (CLL). Fourteen published reports of multigenerational familial CLL were analysed for anticipation, together with 10 previously unreported families with familial CLL, and the difference in disease-free survival between generations was determined. The difference between age at onset for each affected parent-child pair was tested against the null hypothesis that there was no difference in age at onset. The age at onset of the studied cases was also compared with that of the Surveillance Epidemiology and End Results (SEER) Program of the U.S. National Cancer Institute. The median ages at onset in the child and parent generations of all families (51.0 and 72.0 years respectively) were significantly different (P < 0.000001), and the null hypothesis was rejected (P < 0.000001). A significant difference was observed between the ages of onset of the child generation and the SEER population (P < 0.00001), but not between the parent generation and the SEER population. Anticipation characterizes familial CLL. PMID- 11380407 TI - Microvessel overexpression of aquaporin 1 parallels bone marrow angiogenesis in patients with active multiple myeloma. AB - The erythrocyte water channel aquaporin 1 (AQP1) is expressed in multiple absorptive and secretory epithelia including the capillary endothelia. Immunoblot analysis showed that bone marrow biopsies of patients with active multiple myeloma (MM) display significantly higher levels of AQP1 than those from patients with non-active MM, whose values are higher, but to a lesser extent, than those of patients with monoclonal gammopathies of undetermined significance (MGUS). Values of MGUS overlapped those of patients with anaemia as a result of iron or vitamin B12 deficiencies (called 'benign anaemias'). Immunohistochemistry and computerized image analysis of AQP1 highlighted bone marrow microvessels whose area per microscopic field was significantly greater in patients with active MM, and always larger than and closely correlated with the microvessel area when assessed with factor VIII-related antigen/von Willebrand's factor (FVIII-VWF). The intensity of AQP1 expression by microvessels evaluated using image analysis was significantly greater in active than non-active MM and in the latter over MGUS or benign anaemias. It is suggested that, among plasma cell tumours, AQP1 expression is preferentially associated with microvessels of MM and that the highest degree of expression occurs in active MM in step with enhanced angiogenesis, in which AQP1 recognizes more immature neovessels than FVIII-VWF. It may, perhaps, favour angiogenesis in a positive loop and, hence, MM progression, and thus be applied for therapeutic vascular targeting. PMID- 11380408 TI - Thalidomide in multiple myeloma: lack of response of soft-tissue plasmacytomas. AB - Thalidomide is active in patients with refractory myeloma. Seventeen patients (nine men/eight women, median age 73 years) with multiple myeloma (MM) were treated with thalidomide. Fifteen patients had refractory disease and two untested relapse. The median dose of thalidomide was 500 mg (range, 200-800 mg). Nine of the 17 patients (53%) responded. The response rate was significantly higher in patients with no extramedullary disease than in those with soft tissue masses (75% CI: 43-95% versus 0%; P = 0.01)). Of note, no decrease in the size of soft tissue plasmacytomas was observed in all the five patients who had extramedullary involvement. This data suggests that the mechanism of action and effectiveness of thalidomide might depend on the site of the tumour cells. PMID- 11380409 TI - Post renal transplantation human herpesvirus 8-associated lymphoproliferative disorder and Kaposi's sarcoma. AB - Post-transplantation lymphoproliferative disorders (PTLDs) and Kaposi's sarcoma (KS) are immunosuppression-related tumours developing in solid organ transplant patients. Although the Epstein-Barr virus (EBV) is detected in the majority of the PTLDs during the first year after transplantation, the proportion of EBV negative PTLDs has increased in recent years. We report a case of a 17-year-old man who developed severe immune haemolytic anaemia, KS and human herpesvirus 8 (HHV-8)-associated, polymorphic-type PTLD 9 months after allogeneic renal transplantation from his HHV-8-seropositive father. It is suggested that: (i) HHV 8 may be associated with EBV-negative, polymorphous-type PTLD occurring less than 1 year after transplantation, and (ii) PTLD may be listed among other tumours, including KS, Castleman's disease and primary effusion lymphoma (PEL), that are related to HHV-8 infection. PMID- 11380410 TI - Marked decrease in the Epstein-Barr virus positivity rate in nodular sclerosis subtype Hodgkin's disease in Tokyo: trend between 1955 and 1999. AB - We studied the trends of the Epstein-Barr virus (EBV) positivity rate in Hodgkin's disease (HD) between 1955 and 1999. The overall positivity rate gradually decreased over time [12 out of 23 (52%) in 1955-69; 18 out of 39 (46%) in 1970-84; 13 out of 44 (35%) in 1985-99; P = 0.135]. Interestingly, a marked decrease in the rate was noted in the nodular sclerosis (NS) subtype [5 out of 11 (45%); 4 out of 19 (21%); 1 out of 24 (4%); P < 0.01] but not in other classic HD [7 out of 11 (63%); 14 out of 18 (78%); 12 out of 18 (67%); P = 0.775]. Our results indicate that the EBV positivity rate in HD has decreased over time and that this trend reflected the increasing incidence of EBV-negative NS. PMID- 11380411 TI - Diffuse large B-cell non-Hodgkin's lymphoma in a patient with autoimmune lymphoproliferative syndrome. AB - Mutations of Fas or Fas ligand genes result in the autoimmune lymphoproliferative syndrome (ALPS) in humans. We report here a diffuse large B-cell non-Hodgkin's lymphoma occurring in a man with ALPS. Fas-mediated lymphocyte apoptosis was defective in vitro, owing to a mutation within the death domain of the Fas molecule. High-dose methotrexate and doxorubicin-based chemotherapy led to complete remission of lymphoma. PMID- 11380412 TI - Molecular cytogenetic characterization of an acquired minute supernumerary marker chromosome as the sole abnormality in a case clinically diagnosed as atypical Philadelphia-negative chronic myelogenous leukaemia. AB - A case of chronic myelogenous leukaemia (CML) in a 48-year-old man is reported. To the best of our knowledge, this is the first report of a Philadelphia-negative CML with an acquired small supernumerary marker chromosome (SMC) 11 as the sole abnormality. The derivative chromosome 11 was studied in detail using molecular cytogenetic methods; fluorescence in situ hybridization (FISH) using centromere- and region-specific probes for chromosome 11, microdissection, micro-comparative genomic hybridization (micro-CGH) and the recently developed multicolour banding (MCB) technique. The acquired SMC was determined to be a ring chromosome that can be described as r(11)(:p11.2-->q13.1:q14:). PMID- 11380413 TI - Variant Philadelphia translocations in chronic myeloid leukaemia can mimic typical blast crisis chromosome abnormalities or classic t(9;22): a report of two cases. AB - A range of fluorescent in situ hybridization techniques have been used to reveal hidden variant Philadelphia translocations in two cases of Ph-positive chronic phase chronic myeloid leukaemia. In one patient, a highly complex variant Ph translocation affecting four chromosomes had resulted in the formation of structures with the appearance of i(17q) and +8. Misinterpretation of these karyotypes has direct clinical relevance. Our findings illustrate that even established cytogenetic abnormalities may contain cryptic abnormalities beyond the resolution of conventional cytogenetic methods. PMID- 11380414 TI - Long-term outcome of patients surviving for more than ten years following treatment for acute leukaemia. AB - Between 1972 and 1988, 832 consecutive patients were treated for acute leukaemia at St. Bartholomew's Hospital; a retrospective analysis has been conducted to determine the clinical course and outcome for 101 who have survived > or = 10 years following treatment. At a median follow-up of 16 years (range 10-28 years), 86 patients (86 out of 834 total, 11%) were still alive. Long-term follow-up of patients who have survived > or = 10 years following treatment for acute leukaemia revealed that most patients were in normal health, although a significant number of complications had occurred. PMID- 11380415 TI - Older patients with high-risk fungal infections can be successfully allografted using non-myeloablative conditioning in combination with intensified supportive care regimens. AB - Leukaemic patients with advanced disease and severe fungal infections as well as older patients with substantial co-morbidity are usually excluded from conventional allotransplantation because of increased morbidity and mortality. We approached allogeneic transplantation in four patients with a median age of 62 years (one chronic myeloid leukaemia in blast crisis, one high-risk acute myeloid leukaemia (AML) in first complete remission (CR1), one AML in 2nd relapse, one AML in CR2 with pre-existing fungal lung infections (two aspergillus, two mucor) and additional co-morbidity (diabetes n = 2, aortic aneurysm n = 1, arterial sclerosis n = 2) by combining non-myeloablative conditioning with an intensified supportive care regimen, including amphotericin B and 4-12 (median 9) prophylactic granulocyte transfusions from granulocyte colony-stimulating factor (G-CSF)-stimulated volunteer donors. G-CSF was also given to patients until neutrophil recovery. All four patients recovered to a neutrophil count of 0.5 x 109/l after a median of 11.5 d (range 11-13 d). Prophylactic granulocyte transfusions also reduced the need for platelet transfusions and minimized mucositis. All patients were discharged at a median of 25 d (range 18-59 d) and are alive and well after a median follow-up of > 390 d (range 336-417 d) without evidence of leukaemia. Regression of the fungal lesions was documented in three patients, with a slight progression detected by computerized tomography scan of the chest in one patient. We conclude that pulmonary fungal infections are not a contraindication for allogeneic stem cell transplantation, if non-myeloablative conditioning regimens are used in combination with granulocyte transfusions, intravenous amphotericin B and G-CSF. PMID- 11380416 TI - A pilot study of combined immunotherapy with autologous adoptive tumour-specific T-cell transfer, vaccination with CD40-activated malignant B cells and interleukin 2. AB - Most B-cell malignancies are incurable diseases and therefore warrant new therapeutic approaches. In a pilot study, we tested the feasibility and safety of combined immunotherapy consisting of adoptive transfer of autologous tumour specific T cells, low-dose interleukin 2 (IL-2) and a cellular vaccine of CD40 activated plasma cell leukaemia (PCL) cells in a patient who failed tandem repeat stem cell transplantation and idiotype vaccination. Autologous tumour-specific T cells for adoptive T-cell transfer were propagated in vitro by repetitive stimulation with autologous ex vivo CD40-activated PCL cells. CD40-activated PCL cells for vaccination were similarly generated ex vivo by co-culture with CD40 ligand transfectants. Autologous T cells (5 x 108 and 2.5 x 109 for two separate treatment cycles) generated ex vivo and cytotoxic against autologous tumours were infused and well tolerated by the patient. Fever and myalgias were closely related to IL-2 injections and no other adverse effects were observed. A temporary decrease of PCL cells in peripheral blood was seen after the first cycle of adoptive T-cell therapy, tumour cell vaccination and low-dose IL-2. Tumour progression was associated with tumour cells that (1) expressed a complex karyotype, (2) demonstrated loss of MHC class II, and (3) did not induce autologous tumour-specific T-cell lines ex vivo. We demonstrated the safety and feasibility in combining autologous tumour-specific T-cell therapy with low-dose IL-2 and that clinical trials based on the use of CD40-activated autologous tumour cell vaccines are warranted in patients with CD40-activated autologous tumour cells, either as a vaccine or for ex vivo stimulation of autologous T cells. PMID- 11380418 TI - Apoptosis and megakaryocytic differentiation during ex vivo expansion of human cord blood CD34+ cells using thrombopoietin. AB - Thrombopoietin (TPO), the primary regulator of megakaryocytopoiesis, plays important roles in early haematopoiesis. Previously, we have demonstrated that TPO induces a characteristic pattern of apoptosis during ex vivo expansion of cord blood (CB) CD34+ cells. In this study, we have demonstrated that the TPO induced apoptotic cells belong to the megakaryocytic (MK) lineage and that initially expanding MK progenitors declined along with the appearance of TPO induced apoptosis. Human CB CD34+ cells were expanded in serum-free conditions with TPO. Multidimensional flow cytometry using simultaneous measurement of apoptosis and immunophenotyping showed that the TPO-induced apoptotic cells appeared in CD61+ fractions. Immunocytochemical analysis of the fluorescent activated cell-sorted fractions showed that the apoptosis-associated CD44low fraction expressed CD61. Clonogenic assay revealed 7.4 +/- 0.50-fold increase of total megakaryocyte colony-forming units (CFU-MKs) during the initial 9 d. Thereafter, the number of CFU-MKs decreased in parallel with the increase of apoptosis. When the MK colonies were subdivided according to size, the proportion of large colonies progressively decreased, while that of medium and small colonies increased. In particular, from d 6 small colonies became predominant. These results suggested that the MK progenitors matured as they expanded during ex vivo expansion with TPO and then proceeded to apoptosis. PMID- 11380417 TI - A proposed objective way to assess results of randomized prospective clinical trials with acute graft-versus-host disease as an outcome of interest. AB - The assessment of graft-versus-host disease (GVHD) as an end-point in clinical trials requires subjective judgement to distinguish morbidity caused by GVHD from morbidity caused by other complications. We developed a method based on ordinal regression for longitudinal assessment of morbidity involving the skin, liver and gut, regardless of cause as an objective end-point in randomized prospective phase III treatment or prevention trials for which GVHD is an outcome of interest. This method was validated for treatment studies by showing that morbidity was more severe among patients with grade II-IV GVHD than among those who did not have GVHD. We found no differences in morbidity involving the skin, liver and gut after the diagnosis of GVHD in a group of 30 patients who received peripheral blood stem cells and a group of 37 who received marrow in a randomized phase III clinical trial. These preliminary results suggest that objective end points could be used in randomized clinical trials for treatment of GVHD. Further studies will be needed to determine if similar methods could be used in randomized clinical trials for prevention of GVHD. PMID- 11380419 TI - Selective ex vivo expansion of cytomegalovirus-specific CD4+ and CD8+ T lymphocytes using dendritic cells pulsed with a human leucocyte antigen A*0201 restricted peptide. AB - Adoptive transfer of ex vivo-generated cytomegalovirus (CMV)-specific T lymphocytes may be effective in preventing CMV disease in allogeneic haematopoietic stem cell transplantation (HSCT) recipients. We developed a procedure for expansion of CMV-specific T lymphocytes based on the antigen presenting function of donor dendritic cells (DCs), pulsed with a human leucocyte antigen A*0201-restricted pp65 nonamer peptide. CMV-specific T lymphocytes were identified following induction of interferon gamma (IFN-gamma) secretion prompted by peptide exposure. Both CD8+ and CD4+ CMV-specific T lymphocytes were selectively produced in these cultures and showed CMV-restricted cytotoxicity. The simultaneous and selective expansion of CD4+ and CD8+ CMV-specific lymphocytes might be instrumental for more efficient in vivo function of infused CMV-specific lymphocytes. PMID- 11380420 TI - Immune neutropenia associated with anti-human neutrophil antigen-2a (NB1) antibodies following unrelated donor stem cell transplantation for chronic myeloid leukaemia: perpetuation by granulocyte colony-stimulating factor. AB - A case of immune neutropenia following unrelated stem cell transplantation for chronic myeloid leukaemia is described. The neutropenia developed following herpes zoster viral infection and was associated with antibodies to the human neutrophil antigen (HNA)-2a (formerly known as NB1). The neutropenia was prolonged, profound and unresponsive to granulocyte colony-stimulating factor (GCSF). The neutrophil count recovered after GCSF was discontinued. HNA-2a has been reported to be upregulated following GCSF administration. In the present case, it appears that the immune neutropenia may have been perpetuated by GCSF administration. PMID- 11380421 TI - Oxidative stress and erythrocyte damage in Kenyan children with severe Plasmodium falciparum malaria. AB - Anaemia causes significant morbidity in children with Plasmodium falciparum malaria, but the mechanism(s) are unclear. During malarial infection, increased reactive oxygen species (ROS) are generated that may contribute to erythrocyte damage and anaemia. This study measured the concentrations of alpha-tocopherol in plasma and erythrocyte membranes, and the percentage polyunsaturated fatty acid composition (%PUFA) (an indirect marker of ROS damage) in erythrocyte membranes in children with severe P. falciparum malaria from Kilifi, Kenya, and asymptomatic children from the same district. Malarial subjects were stratified into complicated malaria and malaria anaemia. Results demonstrated significant reductions in erythrocyte membrane alpha-tocopherol concentration (1.63 +/- 0.16 versus 3.38 +/- 0.18 micromol/mg protein; P < 0.001) and total %PUFA (30.7 +/- 0.49 versus 32.8 +/- 0.44% P < 0.005) for the malarial subjects (non-stratified) compared with controls. Malarial subjects showed a significant positive correlation between membrane alpha-tocopherol and haemoglobin concentrations (P < 0.005 r = 0.63 complicated malaria group; P < 0.05 r = 0.36 non-stratified data). There were no significant differences in plasma alpha-tocopherol concentration between malaria patients and controls. In conclusion, malarial infection may be associated with oxidative damage and reduced alpha-tocopherol reserve in the erythrocyte membrane, suggesting that local antioxidant depletion may contribute to erythrocyte loss in severe malaria. Erythrocyte membrane alpha-tocopherol appeared a better indicator of ROS exposure than plasma. PMID- 11380422 TI - Growth of Plasmodium falciparum induces stage-dependent haemichrome formation, oxidative aggregation of band 3, membrane deposition of complement and antibodies, and phagocytosis of parasitized erythrocytes. AB - Plasmodium falciparum-parasitized erythrocytes (RBCs) are progressively transformed into non-self cells, phagocytosed by human monocytes. Haemichromes, aggregated band 3 (Bd3) and membrane-bound complement fragment C3c and IgG were assayed in serum-opsonized stage-separated parasitized RBCs. All parameters progressed from control to rings to trophozoites to schizonts: haemichromes, nil; 0.64 +/- 0.12; 5.6 +/- 1.91; 8.4 +/- 2.8 (nmol/ml membrane); Bd3, 1 +/- 0.1; 4.3 +/- 1.5; 23 +/- 5; 25 +/- 6 (percentage aggregated); C3c, 31 +/- 11; 223 +/- 86; 446 +/- 157; 620 +/- 120 (mOD405/min/ml membrane); IgG, 35 +/- 12; 65 +/- 23; 436 +/- 127; 590 +/- 196 (mOD405/min/ml membrane). All increments in rings versus controls and in trophozoites versus rings were highly significant. Parasite development in the presence of 100 micromol/l beta-mercaptoethanol largely reverted haemichrome formation, Bd3 aggregation, C3c and IgG deposition and phagocytosis. Membrane proteins extracted by detergent C12E8 were separated on Sepharose CL-6B. Haemichromes, C3c and IgG were present exclusively in the high molecular-weight fractions together with approximately 30% of Bd3, indicating the oxidative formation of immunogenic Bd3 aggregates. Immunoblots of separated membrane proteins with anti-Bd3 antibodies confirmed Bd3 aggregates that, in part, did not enter the gel. Immunoprecipitated antibodies eluted from trophozoites reacted preferentially with aggregated Bd3. Changes in parasitized RBC membranes and induction of phagocytosis were similar to oxidatively damaged, senescent or thalassaemic RBC, indicating that parasite-induced oxidative modifications of Bd3 were per se sufficient to induce and enhance phagocytosis of malaria-parasitized RBC. PMID- 11380423 TI - Resveratrol, a natural dietary phytoalexin, possesses similar properties to hydroxyurea towards erythroid differentiation. AB - Resveratrol, a natural dietary polyphenol, has been postulated to be implicated in the cardioprotective effect of red wine and the low incidence of breast and prostate cancers among vegetarians and Orientals respectively. This compound inhibits ribonucleotide reductase as does hydroxyurea, the first therapeutic agent used in the treatment of sickle cell disease. Using the human erythroleukaemic K562 cell line as an in vitro model, we show here that 50 micromol/l of resveratrol induced a higher haemoglobin production (sevenfold) in K562 cells than 500 micromol/l of hydroxyurea (3.5-fold). This erythroid differentiation was linked to a dose- and time-dependent inhibition of cell proliferation associated with an equivalent increased expression of p21 mRNA, but with a higher increased level of p21 protein (sixfold) for cells treated with resveratrol than for those treated with hydroxyurea (1.5-fold). We also show that 50 micromol/l of resveratrol and 25 micromol/l of hydroxyurea induced variable but similar enhancements of fetal haemoglobin synthesis in cultured erythroid progenitors for the majority of the sickle cell patients studied. These inductions were linked to, but not correlated with, a variable decrease in erythroid burst-forming unit clone number. Taken together, these results show that resveratrol merits further investigations in sickle cell disease therapy. PMID- 11380424 TI - A novel mutation in the SLC19A2 gene in a Tunisian family with thiamine responsive megaloblastic anaemia, diabetes and deafness syndrome. AB - Thiamine-responsive megaloblastic anaemia (TRMA) syndrome with diabetes and deafness was found in two patients from a Tunisian kindred. The proband was homozygous for a novel mutation, 287delG, in the high-affinity thiamine transporter gene, SLC19A2. We demonstrated that fibroblasts from this patient exhibited defective thiamine transport. These data confirm that the SLC19A2 gene is the high-affinity thiamine carrier and that this novel mutation is responsible for TRMA syndrome. PMID- 11380425 TI - Platelet tests in the prediction of myocardial infarction and ischaemic stroke: evidence from the Caerphilly Prospective Study. AB - A platelet test that is predictive of myocardial infarction (MI) and/or stroke would enable the targeting of anti-platelet drugs towards high-risk patients. The predictive power of several platelet tests for MI and for stroke was examined in 2000 older men in the Caerphilly Cohort Study of Heart Disease, Stroke and Cognitive Decline. The tests were: aggregation to adenosine diphosphate (ADP) in platelet-rich plasma (PRP); aggregation to ADP in whole blood measured using an impedance method and a test of platelet aggregation induced in whole blood by high-shear flow. Around 200 MIs and 100 ischaemic strokes occurred during a 10 year follow-up. Neither primary nor secondary aggregation in PRP was predictive of MI. However, the fifth of men in whom the primary response to ADP was least, showed the highest risk of a subsequent stroke [relative odds (RO) 1.64; 95% confidence interval (CI) 1.12-2.43]. Aggregation in whole blood was not predictive of MI but, again, the fifth of men with the least platelet response showed the highest stroke incidence (RO 1.79; 95% CI 1.06-3.00). Retention of platelets in the high-shear test was not predictive of either event. PMID- 11380426 TI - Endothelial cell prostacyclin synthesis induced by lymphocytes is independent of the membrane fatty acid composition of both cell types and of E-selectin, VCAM-1 or ICAM-1-mediated adhesion. AB - Prostacyclin (PGI2), the main prostanoid in most vascular tissues regulates haemostasis and vascular tone, as well as the proliferation of smooth muscle cells. We have previously reported that lymphocyte contact with endothelium enhances endothelial cell PGI2 output. Here, we demonstrate the specificity of lymphocytes for switching on this response. Co-incubation of human umbilical vein endothelial cells (HUVEC) in serum-free medium with allogeneic peripheral blood lymphocytes (PBL), at a PBL:HUVEC ratio of 9:1, enhanced the basal (HUVEC alone) PGI2 output by 2.5-fold under static conditions, and was not altered in conditions mimicking shear stress. It occurred without previous activation of either cell type and was dependent upon specific interactions with PBL. Indeed, the PGI2 output induced by the co-incubation with resting neutrophils, non activated platelets or latex beads was significantly lower than that induced by PBL. Blocking endothelial cell adhesion molecules (ECAM) E-selectin, vascular cell adhesion molecule-1 (VCAM-1) or intracellular adhesion molecule-1(ICAM-1) did not modify the PBL-induced PGI2 output, although 51Cr-labelled PBL adhesion was significantly decreased with anti-ICAM-1 antibody. Changes in the fatty acid composition of membrane phospholipids induced by incubation with eicosapentaenoic (EPA) or docosahexaenoic acids (DHA) resulted in diminished basal PGI2 output and adhesion of 51Cr-labelled PBL, whereas the PBL-stimulated PGI2 output was not modified. This specific cell-cell interaction represents a new stimulus for PGI2 synthesis that does not primarily involve the ECAM pathway, is independent of cell membrane fatty acid composition and shear stress. This switch-on for PGI2 synthesis, which is induced by lymphocytes, might serve as a protection against atherogenesis. PMID- 11380427 TI - Potency estimation of recombinant factor VIII: effect of assay method and standard. AB - Large potency discrepancies between the chromogenic and one-stage clotting methods have been reported for patients' plasma samples following the infusion of recombinant factor VIII (rFVIII) concentrates. We have investigated the potency estimation of two different full-length rFVIII concentrates using both assay methods relative to both plasma and concentrate standards. Potencies by the chromogenic method were significantly higher (53% and 45%) than potencies by the one-stage clotting method when a plasma standard was used. In contrast, there was no significant potency difference between methods when a concentrate standard was used. Time-course studies into thrombin and activated factor X (FXa) generation, in modified clotting and chromogenic methods, respectively, revealed that the two rFVIII concentrates behaved very similarly to the concentrate standard, whereas the plasma standard showed slightly more rapid thrombin generation and markedly slower FXa generation. The different behaviour of rFVIII and plasma FVIII in the chromogenic method is proposed as the main cause of the methods-based potency discrepancy. The results support the use of a concentrate standard to measure rFVIII in post-infusion plasma. PMID- 11380428 TI - Analysis of the tissue factor pathway inhibitor gene and antigen levels in relation to venous thrombosis. AB - Tissue factor pathway inhibitor (TFPI) inhibits tissue factor-induced coagulation. The major part of TFPI is releasable by heparin. We recently found eight patients with thrombosis and low levels of heparin-releasable TFPI in whom we investigated the TFPI gene for mutations. A transition of G to A coding for Valine264Methionine in the heparin-binding domain was found. The Val264Met polymorphism had an allele frequency of 3% in 96 healthy individuals. A silent polymorphism was identified in TFPI exon IV (T-->C), which does not alter Tyrosine 56. Apart from Val264Met, which was detected in one out of the eight patients, no other mutations in the TFPI gene were found in patients with low heparin-releasable TFPI. Analysis of Val264Met in 317 patients with deep vein thrombosis (DVT) and 292 controls showed no association between Val264Met and DVT. However, a study of total TFPI antigen levels in 122 DVT patients and 126 controls demonstrated an association between TFPI levels and venous thrombosis (P = 0.0001). These results provide evidence for a relationship between venous thrombosis and total TFPI level as a possible risk factor, whereas they do not support a link between DVT and mutations in the nine exons of the TFPI gene. PMID- 11380429 TI - Anti-prothrombin antibodies: assay conditions and clinical associations in the anti-phospholipid syndrome. AB - Anti-phospholipid antibodies (aPL) are associated with an increased risk of thrombosis and recurrent fetal loss. Antibodies to prothrombin (aPT) have been associated with the anti-phospholipid syndrome (aPS). We assessed variations in aPT assay methodology to optimize an aPT method that was used to screen patients with aPS (n = 66). Detection of aPT using enzyme-linked immunosorbent assay was influenced by the concentration of the capture antigen, the microtitre plate type and the buffer system. The combination of gamma-irradiated plates, a phosphate buffered saline buffer and coating antigen of 10 microg/ml prothrombin was the most sensitive. Both serum and citrate samples are suitable for the detection of aPT. Under these conditions aPT IgM but not IgG were found to be associated with thrombosis and/or fetal loss. PMID- 11380430 TI - Antibodies to factor XII and recurrent fetal loss in patients with the anti phospholipid syndrome. AB - Forty female patients with either primary anti-phospholipid syndrome (n = 26) or systemic lupus erythematosus (anti-phospholipid syndrome positive) (n = 14) were investigated for levels of factor XII, the presence of lupus anticoagulant and antibodies to cardiolipin, beta 2-glycoprotein I and factor XII. Twenty-one patients had a history of recurrent fetal loss (> 2, mean = 2.6). Lupus anticoagulant positivity showed a weak association with recurrent fetal loss (odds ratio = 1.1). While there was no association between the presence of antibodies to cardiolipin or beta 2-glycoprotein I with recurrent fetal loss, antibodies to factor XII showed a strong and statistically significant association (odds ratio = 5.4, P = 0.025). PMID- 11380431 TI - The risk of pregnancy-related venous thromboembolism in women who are homozygous for factor V Leiden. AB - The risk of venous thromboembolism (VTE) is increased in pregnancy and during the post-partum period. The absolute risk for pregnancy-related VTE in heterozygous women with the factor V Leiden mutation is approximately 2%, but studies on this risk for homozygous women show conflicting results. In a retrospective family study, we found that the risk of pregnancy-related VTE in women with a symptomatic first-degree relative was 17% per pregnancy (95%CI 4.7-37.4). Anticoagulant prophylaxis during the post-partum period appears to be indicated in asymptomatic homozygous women from symptomatic kindred, whereas this could be decided on an individual basis during pregnancy. PMID- 11380432 TI - Incidence and associations of acute renal failure complicating high-dose intravenous immunoglobulin therapy. AB - In a formal study, we have identified increasing age, pretreatment renal impairment and diabetes mellitus as risk factors for the development of intravenous immunoglobulin-induced renal failure. Identification of these characteristics in potential recipients should alert clinicians to the associated increased risk of this serious complication. PMID- 11380433 TI - Following the course of malaria treatment by detecting parasite lactate dehydrogenase enzyme. PMID- 11380434 TI - Thrombolysis failure: a role for plasminogen activator inhibitor type 1 (PAI-1). PMID- 11380435 TI - Potential strategies for the treatment of plasma exchange-resistant thrombotic thrombocytopenic purpura. PMID- 11380438 TI - Angiogenesis in multiple myeloma. PMID- 11380439 TI - Disseminated Mycobacterium avium intracellulare infection in a patient with AIDS. PMID- 11380440 TI - The hepatologist in the haematologists' camp. PMID- 11380441 TI - The history of folic acid. PMID- 11380442 TI - Hepatic fibrosis plays a central role in the pathogenesis of thrombocytopenia in patients with chronic viral hepatitis. AB - The pathogenesis of thrombocytopenia in chronic hepatitis is not well known. This study evaluated the relationship between liver injury, serum thrombopoietin, splenomegaly and thrombocytopenia in chronic viral hepatitis. Two hundred and nine patients were enrolled, 85 with splenomegaly and 124 without. Thrombocytopenia was present in 71% and 23% of patients with or without splenomegaly respectively. In subjects with low platelet count, those with splenomegaly showed significantly lower platelet numbers than those without splenomegaly. The spleen size correlated with portal hypertension. An inverse correlation between spleen size and platelet count was observed (r = -0.54; P < 0.0001). In patients without splenomegaly, thrombocytopenia was associated with the grade of fibrosis; platelet counts were the highest in patients with fibrosis 0-2, lower in those with grade 3 (P < 0.008) and lowest in those with grade 4 (P < 0.05). These findings were independent of demographic and biochemical characteristics, hepatic necroinflammatory activity, portal hypertension and splenomegaly. Patients with normal platelet counts showed higher thrombopoietin levels than those with low platelet counts (P < 0.0001). An inverse correlation between thrombopoietin levels and fibrosis grade was observed (r = - 0.50; P < 0.0001). Median thrombopoietin levels were 58 and 27 pg/ml for fibrosis grade 0-1 and grade 4 respectively (P < 0.001). These data indicate that advanced hepatic fibrosis, causing an altered production of thrombopoietin and portal hypertension, plays the central role in the pathogenesis of thrombocytopenia in chronic viral hepatitis. PMID- 11380444 TI - Incidence of factor VIII inhibitors in severe haemophilia: the importance of patient age. AB - The potential effect of age at the start of replacement therapy on the development of factor VIII (FVIII) inhibitors was assessed in 62 severe (FVIII < 2 IU/dl) haemophilia A patients who started FVIII therapy at one of two haemophilia centres. Inhibitors were tested on an annual basis. Persistent or high-titre inhibitors were detected in 15 patients (24%). Kaplan-Meier cumulative incidence at 3 years from first FVIII exposure was 41% (95% CI 22-67%) in patients starting therapy before the age of 6 months, 29% (95% CI 13-57%) in patients starting therapy between 6 and 12 months of age, and 12% (95% CI 4-34%) in those starting therapy beyond 1 year of age (P = 0.03). By multivariate analysis, the influence of age was shown to be independent of other variables, including calendar year at the onset of therapy and baseline FVIII plasma levels. In conclusion, patient age at initial treatment appears to influence inhibitor formation. If confirmed, this finding would have a major impact on the management of haemophilia. PMID- 11380443 TI - Polymorphisms in inflammatory cytokines and Fcgamma receptors in childhood chronic immune thrombocytopenic purpura: a pilot study. AB - Inflammatory cytokines and low-affinity Fcgamma receptor (FcgammaR) polymorphisms were investigated in 37 children with chronic immune thrombocytopenic purpura (cITP) and 218 controls. Genotype analysis included common variants in the regulatory regions of cytokines, TNF, LTA, IL1RN, IL1A, IL1B, IL4, IL6 and IL10, and structural variants of the low affinity FcgammaRs, FCGR2A, FCGR3A and FCGR3B. Associations were observed for TNF (P = 0.0032), LTA (P = 0.019), FCGR3A (P = 0.038) and FCGR3B (P = 0.0034). Two combinations of genotypes (TNF and FCGR3A; P = 0.0003, and LTA and FCGR3B; P = 0.011) were significantly associated with cITP. These results provide preliminary evidence that variant genotypes of FcgammaRs and cytokines contribute to cITP pathogenesis. PMID- 11380445 TI - Stable recombinant expression and characterization of the two haemophilic factor VIII variants C329S (CRM(-)) and G1948D (CRM(r)). AB - In haemophilia A, the functional defect at the molecular level of most factor VIII (FVIII) missense mutations remains unknown. Site-directed mutagenesis of B domain-deleted FVIII cDNA (FVIIISQ) was used to introduce two mutations associated with severe cross-reacting material (CRM)-negative (FVIII-C329S) or mild/moderate CRM-reduced (FVIII-G1948D) haemophilia A. Wild-type (FVIIISQ-WT) and variant FVIIISQ proteins were successfully expressed after stable transfection in Chinese hamster ovary (CHO) cells, and partially characterized at the intracellular, molecular and functional levels. Reverse transcription polymerase chain reaction analysis confirmed that both transcription and mRNA processing appeared normal in CHO cells transfected with both the wild-type and two variant constructs. In contrast to FVIIISQ-WT, immunofluorescence analysis of both CRM(-) and CRM(r) variants showed intracellular FVIII accumulation within the rough endoplasmic reticulum, suggesting secretion defects in transfected CHO cells. Immunoblot analysis of the FVIIISQ variant proteins that were secreted showed that they were expressed as mixed populations of uncleaved 170 kDa polypeptides, processed 90 kDa heavy chains and 80 kDa light chains, similar to FVIIISQ-WT. Phenotypic analysis of the B domain-deleted FVIIISQ variants expressed in CHO cells correlated well with the patients' reduced FVIII activity and, in addition, surface plasmon resonance studies demonstrated that both missense mutations were associated with increased rates of A2 domain dissociation following thrombin activation. We conclude that the mutations found are responsible for the haemophilia A phenotype, through intracellular retention and decreased stability of the active cofactor FVIIIa. PMID- 11380446 TI - Deletion of the factor IX gene as a result of translocation t(X;1) in a girl affected by haemophilia B. AB - A balanced de novo translocation t(X;1) is described in a girl with severe haemophilia B. The translocated X was shown cytologically to be preferentially active, and methylation analysis of the DXS255 locus confirmed the skewed X inactivation with the paternal allele being the active one. Cytogenetic and molecular analysis showed that this chromosomal rearrangement led to the deletion of at least part of the factor IX gene. Therefore, the girl was heterozygous for factor IX deficiency and expression of her clinical phenotype was the result of the inactivation of the normal maternal X chromosome. The localization of one of the X chromosome translocation breakpoints in YAC clone 957F9, that was demonstrated to map distally to the factor IX gene, revealed the complexity of this chromosomal rearrangement. PMID- 11380448 TI - The risk of recurrent venous thromboembolism among heterozygous carriers of the G20210A prothrombin gene mutation. AB - The G20210A mutation in the prothrombin gene is associated with an increased risk of a first venous thromboembolic episode; few data are available about the long term risk for recurrent venous thromboembolism and it is not known whether or not carriers of the mutation should be recommended lifelong anticoagulant treatment after the first thrombosis. We investigated 624 patients, referred for previous objectively documented deep venous thrombosis of the legs or pulmonary embolism, to determine the risk of recurrent thromboembolism in heterozygous carriers of the G20210A mutation in the prothrombin gene after the first episode of venous thromboembolism. After exclusion of other inherited (anti-thrombin, protein C, protein S deficiency and factor V Leiden) or acquired (anti-phospholipid antibody syndrome) causes of thrombophilia, 52 heterozygous carriers of the prothrombin mutation were compared with 283 patients with normal genotype. The relative risk for recurrent venous thromboembolism was calculated between groups using a Cox's proportional hazard model. The patients with the prothrombin mutation had a risk for spontaneous recurrent venous thromboembolism similar to that of patients with normal genotype (hazard ratio 1.3; 95% CI, 0.7-2.3). The circumstances of the first event (spontaneous or secondary) did not produce any substantial variation in the risk for recurrence. In conclusion, the carriers of the prothrombin mutation should be treated with oral anticoagulants after a first deep venous thrombosis for a similar length of time as patients with a normal genotype. PMID- 11380447 TI - Complexes of anti-prothrombin antibodies and prothrombin cause lupus anticoagulant activity by competing with the binding of clotting factors for catalytic phospholipid surfaces. AB - We investigated the mechanism by which anti-prothrombin antibodies cause lupus anticoagulant (LAC) activity. Addition of affinity-purified anti-prothrombin antibodies from LAC-positive plasma samples (alpha-FII-LAC+) to normal plasma induced LAC activity. Upon increasing the phospholipid concentration, LAC activity was neutralized. Addition of purified alpha-FII-LAC+ to normal plasma strongly inhibited factor Xa formation. No inhibition was measured when alpha-FII LAC+ were added to prothrombin-deficient plasma or when purified anti-prothrombin antibodies from LAC-negative plasma samples (alpha-FII-LAC-) were added. When a combination of prothrombin and alpha-FII-LAC+ was added to the purified clotting complex, a strong inhibition of factor Xa and IIa formation was seen. The alpha FII-LAC+ alone or a combination of prothrombin and alpha-FII-LAC- did not show inhibition. Ellipsometry studies showed that, in the presence of alpha-FII-LAC+, the affinity of prothrombin for a phospholipid surface increased dramatically, whereas a much lower increase was observed with alpha-FII-LAC-. Our results show that complexes of prothrombin and anti-prothrombin antibodies with LAC activity inhibit both prothrombinase and tenase. The antibodies increase the affinity of prothrombin for the phospholipid surface, thereby competing with clotting factors for the available catalytic phospholipid surface, a mechanism similar to that of anti-beta2-glycoprotein I antibodies. PMID- 11380449 TI - A study of Protein S antigen levels in 3788 healthy volunteers: influence of age, sex and hormone use, and estimate for prevalence of deficiency state. AB - Total Protein S (tPS) and free Protein S (fPS) antigen levels were measured in 3788 healthy blood donors. Men had higher levels of both parameters than women (P < 0.001). Age had no effect on tPS in men, although there was a slight reduction in fPS levels with increasing age. In women increasing age was associated with a significant increase in tPS levels (P < 0.001) but had no effect on fPS after adjustment for menopausal state. Oral contraceptive pill (OCP) use significantly lowered tPS but had no effect on fPS. In post-menopausal women, hormone replacement therapy (HRT) use had no statistically significant effect on either tPS or fPS. Donors with tPS or fPS levels in the lowest percentile (n = 56) were retested; only nine with repeat low levels were identified, eight of whom had persistently low levels over a 4-7-year follow-up. Acquired deficiency was excluded. When possible, family studies were performed, leading to an estimate of prevalence of familial PS deficiency of between 0.03% and 0.13% in the general population. PMID- 11380450 TI - Symptomatic type 1 protein C deficiency caused by a de novo Ser270Leu mutation in the catalytic domain. AB - Heterozygosity for a C8524T transition in the protein C gene converting Ser270(TCG) to Leu(TTG) in the protease domain was identified in a family with venous thrombosis. The mutation was associated with parallel reduction in plasma levels of protein C anticoagulant activity and protein C antigen, which is consistent with a type 1 deficiency. Transient expression of mutant protein C cDNA in human kidney 293 cells and analysis of protein C antigen in culture media and cell lysates showed that the secretion of mutant protein compared with wild type protein was reduced by at least 97% while the intracellular content of mutant and wild-type protein was similar. Northern blot analysis of total mRNA from transfected cells showed no reduction of the mutant protein C mRNA compared with wild-type protein C mRNA. Collectively, these results indicate that the Ser270Leu mutation in the affected family caused the plasma protein C deficiency and are consistent with a disease mechanism that involves synthesis of mutant protein followed by intracellular degradation before its secretion into the extracellular space. The mutation was not present in the parents of the proband, suggesting a de novo mutation. Non-paternity was excluded after the analysis of three intragenic protein C polymorphisms and six dinucleotide repeat allele sets located in five different chromosomes. PMID- 11380451 TI - Ten years of prophylactic treatment with fresh-frozen plasma in a child with chronic relapsing thrombotic thrombocytopenic purpura as a result of a congenital deficiency of von Willebrand factor-cleaving protease. AB - We report the results of 10 years of prophylactic fresh-frozen plasma (FFP) infusion therapy in a 14-year-old girl with chronic relapsing thrombotic thrombocytopenic purpura (TTP), in whom a severe congenital von Willebrand factor (VWF)-cleaving protease deficiency has been documented. Severe haemolytic crises triggered by infections were prevented and her present renal and neurological functions have been preserved. Sequential measurements of protease activity and platelet count after FFP infusion led us to conclude tentatively that 5% may be sufficient to degrade very large and adhesive VWF multimers. PMID- 11380452 TI - Novel Y283C mutation of the A subunit for coagulation factor XIII: molecular modelling predicts its impaired protein folding and dimer formation. AB - In an Italian patient with severe factor XIII deficiency, a novel mutation, Y283C (TAT to TGT), was identified heterozygously by nucleotide sequencing analysis in exon VII of the gene for the A subunit. The presence of this mutation was confirmed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis in the proband and his brother. Molecular modelling predicts that the mutant molecule would be misfolded. It is probable that the impaired folding of the mutant Y283C A subunit led to its instability, which is at least in part responsible for the factor XIII deficiency of this patient. PMID- 11380453 TI - High concentrations of coagulation factor VIII and thrombosis: Is the factor VIII binding domain of von Willebrand factor implicated? AB - The possibility that high factor VIII (FVIII) levels in thrombosis patients is principally explained by a gain of function in the FVIII-binding domain of von Willebrand factor (VWF), arising from amino acid substitution(s) or polymorphism(s), was investigated. Exons 18-24 of the VWF gene were sequenced in 13 thrombosis patients with high FVIII (> 1.50 IU/ml). No novel mutations were found. Four known polymorphisms were detected: G2615A and C2635T (Ex18), G2805A (Ex20) and G3130A (Ex22). Their frequencies showed no significant differences in a thrombosis vs. control cohort. The data suggest that amino acid substitutions/polymorphisms in the VWF-FVIII-binding domain are not the principal explanation for high FVIII in thrombosis patients. PMID- 11380454 TI - Chronic disseminated intravascular coagulation after surgery for abdominal aortic aneurysm: clinical and haemostatic response to dalteparin. AB - A 77-year-old man developed chronic disseminated intravascular coagulation (DIC) after surgical repair of a large infrarenal aortic aneurysm. Self-administered subcutaneous dalteparin therapy (5000 units o.d.) led to rapid relief of symptoms and sustained improvements in his platelet count and fibrinogen level; activation of coagulation and fibrinolysis appeared to be relatively unaffected. Long-term treatment with low-molecular-weight heparin can provide good symptomatic control of chronic DIC associated with abdominal aortic aneurysm. PMID- 11380455 TI - Red cell antibodies in patients with homozygous sickle cell disease: a comparison of patients in Jamaica and the United Kingdom. AB - The transfusion history and frequency of red cell antibodies in patients with homozygous sickle cell (SS) disease have been compared in 190 subjects from the Jamaican cohort study and 37 patients attending a sickle cell clinic in Manchester, England. The proportion of patients transfused did not differ between the groups although the number of units transfused and the frequency of red cell antibodies were significantly greater in the Manchester group. Immune antibodies occurred in three Jamaicans (2.6% of those transfused) and 16 UK subjects (76% of those transfused). Multiple antibodies occurred in 10 (63%) UK subjects but in no Jamaicans. Indications for transfusion also differed between the groups, Jamaican patients typically receiving 1-2 units for acute anaemia or acute chest syndrome, whereas UK patients frequently had multiple transfusions in preoperative exchange or prophylaxis programmes. The greater red cell alloimmunization among UK patients probably reflects both the greater use of transfusion and the disparity between donor and recipient populations in the UK. PMID- 11380456 TI - 16Cys encoded by the RHce gene is associated with altered expression of the e antigen and is frequent in the R0 haplotype. AB - Serological observations have suggested that numerous D, many e (especially in Blacks), several E, and rare c variants exist within the Rh blood group system. The molecular basis for expression of many of these variants has been elucidated. This study describes five unrelated Caucasians whose red blood cells reacted with polyclonal anti-e but did not react with some monoclonal anti-e, which suggested that they carried a variant e antigen. Molecular investigation revealed the presence of a 48G-->C change (encoding cysteine instead of tryptophan at amino acid 16) in their RHce gene. No other differences were found, which suggests that amino acid residues located in the first transmembrane region can affect expression of the e antigen, whose critical residues are on the predicted fourth external loop of the protein. This polymorphism has not previously been observed because polyclonal anti-e does not distinguish this variant from wild type. This position is polymorphic in RHce alleles and the presence of the 48C nucleotide is often found in the R0 (Dce) haplotype. PMID- 11380457 TI - Two new alleles of the RHCE gene in Black individuals: the RHce allele ceMO and the RHcE allele cEMI. AB - Six unrelated individuals of Afro-Caribbean origin, whose red cells have a marked reduction of the Rhe antigen expression, have been identified. All exhibited the same serological profile with anti-e monoclonal antibodies and lacked expression of the high frequency e-related antigen hrS. Transcripts and genomic analysis showed that these phenotypes resulted from the presence of two new RHCE alleles, ceMO and cEMI. The ceMO allele corresponded to a RHce gene carrying a G667T mutation (exon 5) and was detected at the homozygous state in sample 1 and at the heterozygous state in samples 2-6. The G667T mutation resulted in a Val223Phe substitution on the Rhce polypeptide, in close proximity to Ala226 (e-antigen polymorphism), which might account for the altered expression of e. The ceMO allele is also associated with the lack of expression of the hrS antigen. The absence of the hrS antigen expression may have implications in transfusion as hrS negative individuals may develop clinically significant antibodies. The cEMI allele corresponded to a silent RHE allele carrying a nine nucleotide deletion within exon 3 and was detected at the heterozygous state in sample 2. This deletion resulted in a shortened polypeptide of 414 residues (instead of 417) that was absent (or severely reduced) at the red cell surface, as the E antigen was undetectable using serology and Western blot analysis with anti-E reagents. In DNA-based polymerase chain reaction genotyping for RHE determination, the cEMI allele provided a false positive result as the cells carrying this allele are serologically phenotyped as E-negative. The incidence of this allele in the Black population is unknown but, as shown already for D genotyping, one must exercise caution when genotyping is performed to detect the e/E polymorphism. PMID- 11380458 TI - Flow cytometric analysis of the association between blood group-related proteins and the detergent-insoluble material of K562 cells and erythroid precursors. AB - The linkage between blood group-related cell surface proteins and the detergent insoluble material (DIM) was estimated by flow cytometry using a panel of specific monoclonal antibodies (mAbs) as a comparison of the antibody-binding capacity of intact and Triton-X100-treated cells. Studies were performed with K562 cells expressing endogenous or recombinant proteins and with human erythroid progenitors during their proliferation and differentiation in vitro. Glycophorin C (GPC) was found to be Triton-insoluble in both cellular models. When expressed (erythroid progenitors), Band 3 remained Triton-insoluble. Glycophorin A (GPA), however, behaved as Triton-soluble or insoluble according to the absence (K562) or the presence (erythroid progenitors) of Band 3 respectively. Comparison of the cellular models regarding the proteins that compose the Rh complex also indicated that Rh(D), RhAG and CD47 were resistant to Triton extraction in cells lacking Band 3. Similarly, RhAG and CD47 remained predominantly Triton-insoluble in K562 cells and early progenitors before Rh and Band 3 expression. Further analysis showed that the Kell protein was DIM-associated. In contrast, CD99 and DARC (Fy) proteins were not, or were very poorly, DIM-associated. Additionally, the adhesion molecules CD44 and Lu were completely or partially resistant to detergent extraction respectively. Deletion of the Lu cytoplasmic tail or its replacement by the cytoplasmic domain of GPC resulted in significant increase or decrease of the Triton solubility of the transfected proteins respectively. These data suggest that Triton insolubility of Lu results in part from direct attachment of its cytoplasmic tail with the cytoskeleton. We assume that this method should provide a useful tool to map interaction sites localized in the cytoplasmic domain of recombinant transmembrane proteins. PMID- 11380459 TI - Band 3 Cape Town (E90K) causes severe hereditary spherocytosis in combination with band 3 Prague III. AB - Hereditary spherocytosis (HS) is an inherited haemolytic anaemia, characterized by spheroidal, osmotically fragile red blood cells. This disorder exhibits heterogeneity in terms of both clinical severity and underlying molecular defect. We have studied a South African Cape Coloured individual with severe HS owing to a band 3 deficiency caused by two mutations, occurring in trans, in the band 3 gene: a novel variant that we have designated band 3 Cape Town and a previously described mutation, band 3 Prague III. Analysis of erythrocyte membrane proteins indicated a deficiency of both band 3 and protein 4.2, as well as a decreased functional capacity of band 3 to transport anions. Band 3 Cape Town is defined by a GAG-->AAG point mutation at codon 90, substituting a glutamic acid with a lysine in the cytoplasmic domain of the molecule, while band 3 Prague III is a codon 870 CGG-->TGG point mutation, replacing an arginine with a tryptophan in the transmembrane region of band 3. mRNA is transcribed from both mutant alleles, implying that mutant proteins are synthesized, but are either degraded prior to membrane incorporation or insertion is impaired. We conclude that the combination of these two mutations exacerbated the clinical presentation of the proband. PMID- 11380460 TI - Polymerase chain reaction protocols for alpha globin haplotype polymorphisms. AB - Polymerase chain reaction protocols were designed specifically to amplify regions of the alpha globin complex that contain the nine common polymorphic haplotyping sites. These reactions provided a quicker and more sensitive approach to determining alpha globin haplotypes than Southern blotting methods. PMID- 11380461 TI - Lack of CD40-dependent B-cell proliferation in B lymphocytes isolated from patients with persistent polyclonal B-cell lymphocytosis. AB - Persistent B-cell lymphocytosis (PPBL) is a haematological disorder diagnosed primarily in adult female smokers that is characterized by a polyclonal increase in peripheral blood B lymphocytes and a moderate elevation of serum IgM. B lymphocyte-associated cellular abnormalities, such as the occurrence of multi lobed nuclei, increased bcl2/Ig gene rearrangements and the identification of an extra long-arm chromosome (i3)(q10) in the B-cell population, indicate that PPBL could be part of a multi-step process leading to the emergence of a malignant B lymphoproliferation. However, the resulting impact on cellular functional properties remains to be elucidated. Our goal was to address that aspect via the study of B-cell activity following stimulation through CD40, a key molecule of the tumour necrosis factor receptor superfamily involved in B lymphocyte development. In contrast to normal B cells, PPBL B lymphocytes were unable to respond to the proliferative signal delivered in vitro by CD40, indicating a defect in the CD40 activation pathway. Polymerase chain reaction amplification and sequencing of the receptor as well as FACScan analysis of patient B lymphocytes dismissed the possibility of a defect in either CD40 structure or expression. Moreover, Western blot analysis of tyrosine phosphorylation, an early event in the CD40-signalling cascade, was similar in patients and controls, leading to the conclusion that the defect affecting B lymphocytes in PPBL patients is probably located downstream of that signalling cascade. PMID- 11380462 TI - Bcl-2 and Bcl-x expression in the CD34+ cells of aplastic anaemia patients: relationship with increased apoptosis and upregulation of Fas antigen. AB - Aplastic anaemia (AA) is a syndrome of haemopoietic failure involving increased apoptosis in stem cells. AA CD34+ cells often have upregulated Fas antigen, but this does not explain the increased apoptosis in all patients. To examine whether abnormal expression of the apoptotic modulators Bcl-2 and Bcl-x is involved in increased apoptosis in the CD34+ cells of patients, we examined cells from 19 AA patients and 18 normal controls by triple staining for CD34, Bcl-2 or Bcl-x, together with 7-amino actinomycin D to determine viability or with staining for Fas antigen. We confirmed increased apoptosis of CD34+ cells in patients. All CD34+ cells in patients and controls expressed Bcl-2 and Bcl-x with no significant difference between the groups. In patients, viability of CD34+/Bcl 2hi cells was similar to that of CD34+/Bcl-2lo cells, but CD34+/Bcl-xhi cells were significantly more viable than CD34+/Bcl-xlo cells. CD34+ cells from AA patients expressed upregulated Fas antigen, but this did not correlate with Bcl-2 or Bcl-x expression. These results suggest a more significant role for Bcl-x as an anti-apoptotic regulator in CD34+ cells in AA than Bcl-2. The induction of death by Fas antigen may bypass the anti-apoptotic effect of Bcl-2 and Bcl-x in CD34+ cells in AA. PMID- 11380463 TI - Comparison of 'sequential' versus 'standard' chemotherapy as re-induction treatment, with or without cyclosporine, in refractory/relapsed acute myeloid leukaemia (AML): results of the UK Medical Research Council AML-R trial. AB - This aim of the acute myeloid leukaemia (AML)-R trial was to compare sequential (Seq) ADE (cytarabine, daunorubicin, etoposide) with standard (Std) ADE as remission re-induction treatment and to assess any benefit of cyclosporine (CSA) as a multidrug resistance modulator in refractory/relapsed AML patients. Seq ADE, based on the concept of Timed Sequential Therapy, comprised the same drugs as Std ADE but given at higher doses and in a different sequence. Between 1992 and 1997, 235 patients with relapsed (175) and refractory (60) AML were entered: 170 were randomized between Std versus Seq ADE and 213 between CSA versus no CSA. CSA was initially given at a dose of 5 mg/kg/d and increased to 10 mg/kg/d in the latter part of the trial. Overall, the complete remission (CR) rate was 43%, with Std ADE being significantly better than Seq ADE (54% versus 34%, P = 0.01). CR rates did not differ between the CSA and no CSA arms (41% versus 45%, P = 0.6). Overall, 3 year disease-free survival (DFS) of remitters was 16%, with a relapse risk of 70%. DFS was not significantly different between the chemotherapy or the CSA arms. Overall, 3 year survival was 8%. Survival with Std ADE was significantly better than with Seq ADE (12% versus 6%, P = 0.03). CSA did not affect overall survival, except in patients > or = 60 years, who fared worse on CSA (P = 0.0003). No difference in haematological toxicity between the chemotherapy or CSA arms was seen. Survival was better with longer first CR duration (P < 0.0001). We conclude that Std ADE was superior to Seq ADE for CR achievement and survival, with no benefit with CSA, at the doses used in this study. PMID- 11380464 TI - Patterns of failure with increasing intensification of induction chemotherapy for acute myeloid leukaemia. AB - Patterns of failure were studied in two consecutive randomized trials of intensified induction therapy carried out by the Australian Leukaemia Study Group (ALSG) between 1984 and 1991 to determine the impact of dose intensification. Patients received standard dose cytarabine and daunorubicin (7-3), 7-3 plus etoposide (7-3-7) or 7-3 plus high-dose cytarabine (HIDAC-3-7) chemotherapy. Patients with FAB M3 morphology were excluded. Time to failure (TTF) was defined as the time from randomization to induction death or removal from study for non responders, or to relapse or death in complete response (CR) for complete responders. An estimated 86% of 470 de novo patients with acute myeloid leukaemia failed within 10 years of randomization, as a result of death in induction in 17% of the randomized patients, failure to achieve CR in a further 17%, relapse in 44% and death in CR in 8% of patients. An estimated 66% of patients failed as a result of refractory disease or relapse within that period (disease-related failures). Multifactor analysis identified age and peripheral blast count as the most significant pretreatment factors associated with overall TTF. These factors, together with cytogenetics, were significantly associated with disease-related failures. High-dose cytarabine in induction significantly decreased the disease related failure rate as did allogeneic transplantation in first CR. The impact of high-dose cytarabine did not depend on the cytogenetic risk group. PMID- 11380466 TI - Relationship of p15 and p16 gene alterations to elevated dihydrofolate reductase in childhood acute lymphoblastic leukaemia. AB - The downstream effects of p15 and p16 gene deletions and loss of transcripts on dihydrofolate reductase (DHFR) were examined in 63 B-precursor (BP) acute lymphoblastic leukaemia (ALL) samples. p15 and/or p16 gene deletions were seen in 6% and 8%, respectively, of BP-ALL samples; however, losses of p15 and/or p16 transcripts were seen in 26 out of 63 (41%) samples. Loss of p15 transcripts (36.5%) exceeded that for p16 (17.5%). For the 26 BP-ALLs that lacked p15 and/or p16 transcripts, only six (23%) exhibited low levels of DHFR by flow cytometry assay with Pt430, a fluorescent anti-folate. Conversely, 18 out of 37 (49%) BP ALL samples with intact p15 and/or p16 genes and transcripts showed low levels of DHFR (P = 0.04). In p15- and p16-null K562 cells transfected with a tetracycline inducible p15 cDNA construct, induction of p15 transcripts and protein was accompanied by decreased growth rates, decreased S-phase fraction, decreased retinoblastoma protein phosphorylation, and markedly reduced levels of DHFR transcripts and protein. Collectively, our results suggest that losses of p15 and/or p16 gene expression result in elevated levels of DHFR in BP-ALL in children. However, additional downstream factors undoubtedly also contribute to elevated levels of this enzyme target. PMID- 11380465 TI - Acute myeloid leukaemia M0: haematological, immunophenotypic and cytogenetic characteristics and their prognostic significance: an analysis in 241 patients. AB - Haematological, immunophenotypic and cytogenetic characteristics were analysed in 241 patients with acute myeloid leukaemia (AML) M0, including 58 children. Children < 3 years and adults between 60 and 70 years of age were most frequently affected. Immunophenotyping showed a heterogeneous phenotype. Anti myeloperoxidase was positive in about half of the patients. Cytogenetic data were available from 129 (54%) patients. A normal karyotype was found in only 24%. Most of the abnormalities were unbalanced and the chromosomes 5, 7, 8 and 11 were the most frequently affected. Survival data were available from 152 treated patients (63%). The median overall survival for all patients was 10 months, 20 months for children (n = 36), 10 months for the young adult group (n = 50) and 7 months for the elderly patients (n = 66) (P = 0.09). Karyotype was not a prognostic factor influencing survival. AML M0 shows the immunological characteristics of early progenitor cells, but the expression of the different markers and cytogenetic abnormalities is heterogeneous. The prognosis is poor compared with other de novo AML and similar to that of AML with multilineage dysplasia or AML following myelodysplastic syndromes. PMID- 11380467 TI - Two consecutive immunophenotypic switches in a child with immunogenotypically stable acute leukaemia. AB - A 12-year-old girl presented with a CD33+ precursor B-acute lymphoblastic leukaemia (ALL) and seemed to respond well to ALL treatment. However, 2 weeks after diagnosis her leucocyte count rose rapidly with a predominance of myeloid blasts with M5b morphology and CD19+ myeloid immunophenotype. Acute myeloid leukaemia (AML) treatment was started and remission was achieved after one course of chemotherapy; the AML treatment was continued for 6 months. Two months after cessation of chemotherapy, the patient developed a bone marrow relapse, this time with an undifferentiated blast morphology and a precursor B immunophenotype. Molecular analysis of the immunoglobulin and T-cell receptor genes showed several clonal gene rearrangements at diagnosis: two IGH, two IGK and two TCRD gene rearrangements. All rearrangements were also detected during the AML phase of the disease, suggesting a phenotypic shift of the same leukaemia. At relapse, 8 months later, all rearrangements were preserved except for one TCRD (Vdelta2 Ddelta3) rearrangement. The first phenotypic shift in the genotypically stable leukaemia was remarkably fast. The most probable explanation for our observations is an oncogenic event in an undifferentiated haematopoietic progenitor clone, with a highly versatile phenotype. PMID- 11380468 TI - Cytogenetic findings and their clinical relevance in myelofibrosis with myeloid metaplasia. AB - The prognostic significance of bone marrow cytogenetic lesions in myelofibrosis with myeloid metaplasia (MMM) was investigated in a retrospective series of 165 patients. An abnormal karyotype was demonstrated in 57% of patients. At diagnosis (n = 92), 48% of the patients had detectable cytogenetic abnormalities, and clonal evolution was frequently demonstrated in sequential studies. More than 90% of the anomalies were represented by 20q-, 13q-, +8, +9, 12p-, and abnormalities of chromosomes 1 and 7. Of these, 20q-, 13q- and +8 were the most frequent sole abnormalities, each occurring in 15-25% of the abnormal cases. Trisomy 9 and abnormalities of chromosomes 1 and 7 were equally prevalent but were usually associated with additional cytogenetic lesions. Chromosome 5 abnormalities were infrequent but were over-represented in the group of patients exposed to genotoxic therapy. In a multivariate analysis that incorporated other clinical and laboratory variables, the presence of an abnormal karyotype did not carry an adverse prognosis. Instead, +8, 12p-, advanced age and anaemia were independent prognostic determinants of inferior survival. In particular, survival was not adversely affected by the presence of either 20q- or 13q-. PMID- 11380469 TI - Gemcitabine as a single agent in the treatment of relapsed or refractory low grade non-Hodgkin's lymphoma. AB - A multicentre phase II trial was conducted to evaluate the efficacy and toxicity of gemcitabine in patients with refractory or relapsed indolent non-Hodgkin's lymphoma. Thirty-six patients were enrolled onto the study, including 11 cases of mantle cell lymphoma (MCL), 10 cases of chronic lymphocytic leukaemia (CLL)/lymphocytic lymphoma, nine cases of follicular lymphoma, four cases of lymphoplasmacytic lymphoma and two cases of T-cell lymphoma. Gemcitabine 1 g/m(2) was administered as a 30-min infusion on d 1, 8 and 15 of a 28-d schedule, up to a maximum of six cycles. Complete responses were observed in two patients with MCL, and partial responses were observed in seven patients, including three patients with CLL/lymphocytic lymphoma, two patients with T-cell lymphoma, one patient with MCL and one patient with follicular lymphoma. Minor responses were observed in three patients, including two patients with MCL and one patient with CLL. The median duration of response was 150 d and the overall progression-free survival was 342 d. Haematological toxicity was observed as grade 3-4 leucopenia in 12 patients (33%) and grade 3-4 thrombocytopenia in 18 patients (50%). Severe non-haematological toxicity included one case of fatal veno-occlusive disease, one case of thrombotic microangiopathy leading to terminal renal failure, one case of capillary leak syndrome, one case of myocardial infarction and drug induced fever in two patients. These data suggest that gemcitabine displays activity in patients with MCL and CLL/lymphocytic lymphoma. Haematological toxicity was frequent in these heavily treated patients. Severe non haematological toxicity was significant and should be taken into account in the design of future trials. PMID- 11380470 TI - Interferon alpha and zidovudine therapy in adult T-cell leukaemia lymphoma: response and outcome in 15 patients. AB - Adult T-cell leukaemia lymphoma (ATLL) is an aggressive disease caused by the human T-lymphotropic virus 1 (HTLV-I) with a short survival. Responses to interferon alpha (IFN-alpha) and zidovudine (AZT) have been documented but not with long-term follow-up. We treated 15 ATLL patients with IFN and AZT. Eleven patients had acute ATLL, two had lymphoma and two smouldering ATLL, with progression. The main features were: organomegaly (14), skin lesions (10), high white blood cell (WBC) count (11) and hypercalcaemia (9). Eleven patients had previously received chemotherapy and one had received an autograft. At the time of the study, seven patients had progressive disease and eight were in partial or complete clinical remission. Responses (PR) lasting 2+ to 44+ months were seen in 67%; 26% did not respond (NR) and one patient was not evaluable. Hypercalcaemia predicted a poor outcome but differences were not significant. Eight of the 15 patients have died 3-41 months from diagnosis. Median survival for the 15 patients was 18 months. Survival of the NR ranged from 4 to 20 months; six PR patients are alive 8-82 months from diagnosis. The differences in survival between NR (median: 6 months) and PR (55% of patients alive at 4 years) were statistically significant (P = 0.002). In conclusion, IFN and AZT improves the outcome of ATLL patients and helps maintain responses. PMID- 11380471 TI - Class switch recombination was specifically targeted to immunoglobulin (Ig)G4 or IgA in Hodgkin's disease-derived cell lines. AB - In T cell-dependent immune responses, class switch recombination occurs in germinal centres. There is now evidence that Hodgkin/Reed-Sternberg cells are derived from germinal centre B cells. Cytokines specifically determine the direction of class switching, i.e the isotype of the new antibodies. We performed restriction analyses and polymerase chain reaction on the immunoglobulin heavy chain loci for five Hodgkin's disease-derived B-cell lines and one Hodgkin's disease-derived T-cell line in order to analyse class switch recombination. In all the B-cell lines, class switch recombination had been targeted to Calpha4 or Calpha1/2. This showed that cell-line precursors had undergone class switching, probably under the influence of TH2 or TH3 cell-derived cytokines. Deletions comprising several constant region genes were observed in cell lines L428, L1236, L591 and KMH2. Karyotype analyses of two of these revealed translocational breakpoints within the immunoglobulin heavy chain gene locus. Our data support the view that a chromosomal instability may occur during class switch recombination in Hodgkin/Reed-Sternberg cells causing chromosomal breaks. Thus, as in other germinal centre B cell-derived lymphomas, the immunoglobulin gene locus may be frequently involved in structural chromosomal aberrations in Hodgkin's disease. PMID- 11380472 TI - In multiple myeloma, only a single stage of neoplastic plasma cell differentiation can be identified by VLA-5 and CD45 expression. AB - The nature of the proliferating fraction in myeloma is still not known and understanding the characteristics of this fraction is central to the development of effective novel therapies. However, myeloma plasma cells typically show a very low rate of proliferation and this complicates accurate analysis. Although the level of CD45 and/or VLA-5 has been reported to identify proliferating 'precursor' plasma cells, there are discrepancies between these studies. We have therefore used a rigorous sequential gating strategy to simultaneously analyse cycle status and immunophenotype with respect to CD45, VLA-5 and a range of other integrin molecules. In 11 presentation myeloma patients, the proliferative fraction was distributed evenly between CD45+ and CD45- cells, however, cycling plasma cells were consistently VLA-5-. There was close correlation between the expression of VLA-5 and a range of other integrin molecules (CD11a, CD11c, CD103), as well as the immunoglobulin-associated molecules CD79a/b (Spearman, n = 10, P < 0.0001). In short-term culture, cells that were initially VLA-5-showed increasing VLA-5 expression with time. However, simultaneous analysis of the DNA binding dye 7-amino-actinomycin D demonstrated that this was not as a result of differentiation, as VLA-5+ plasma cells were all non-viable. This was confirmed in freshly explanted plasma cells from nine patients. Discrete stages of plasma cell differentiation could not be distinguished by the level of CD45 or VLA-5 expression. The results indicate that there is a single stage of plasma cell differentiation, with the phenotype CD38+CD138+VLA-5-. These findings support the hypothesis that neoplastic bone marrow plasma cells represent an independent, self-replenishing population. PMID- 11380473 TI - Detection of reciprocal fusion 5'-BCL6/partner-3' transcripts in lymphomas exhibiting reciprocal BCL6 translocations. AB - It has been believed that replacement of the endogenous promoter and the non coding first exon of the BCL6 gene by a sequence derived from the translocational partner gene is a main mechanism of the BCL6 dysregulation resulting from translocation. In this study, we found that reciprocal BCL6 translocation led to the expression of not only the 5'-partner/BCL6-3' fusion transcripts but also the 5'-BCL6/partner-3' fusion transcripts, suggesting that reciprocal 5'-BCL6/partner 3' fusion genes are transcriptionally active. These findings raise the possibility that reciprocal BCL-6 translocation may lead to dysregulation of the partner gene as well as the BCL6 gene. PMID- 11380474 TI - Donor interleukin 1 receptor antagonist genotype associated with acute graft versus-host disease in human leucocyte antigen-matched sibling allogeneic transplants. AB - Interleukin 1 (IL-1) is involved in various autoimmune and inflammatory diseases. IL-1 receptor antagonist (IL-1Ra) is the naturally occurring antagonist to IL 1alpha and -1beta. Polymorphisms of IL-1beta have been associated with variations in IL-1beta production (nucleotides +3953 and -511). A variable number tandem repeat (VNTR) polymorphism in the IL-1Ra gene has been associated (allele 2) with increased IL-1Ra production. We examined these polymorphisms in human leucocyte antigen (HLA)-matched allogeneic bone marrow transplant patients and donors. IL 1Ra VNTR (allele 2) in the donor genotype was more frequent with milder acute graft-versus-host disease (aGvHD) grades 0-II (29 out of 59 transplants) than severe GvHD grades III-IV (2 out of 18 transplants) (P = 0.0032). This association was confirmed in a subgroup with cyclosporine monotherapy prophylaxis: donor possession of allele 2 was again associated with milder aGvHD, grades 0-II (19 out of 38 transplants), than grades III-IV (1 out of 14) (P = 0.0042) transplants. No association was found between the IL-1beta -511 or IL 1beta +3953 polymorphism and severity of GvHD. Recipient IL-1Ra VNTR genotype (allele 2) showed a strong trend towards association with aGvHD severity (P = 0.0697). Thus, the donor genotype for the IL-1Ra polymorphism has an apparent protective role against acute GvHD following transplantation and may be an additional factor for individual risk assessment for complications, including GvHD, post transplant. PMID- 11380475 TI - Molecular quantification of viral load in plasma allows for fast and accurate prediction of response to therapy of Epstein-Barr virus-associated lymphoproliferative disease after allogeneic stem cell transplantation. AB - Epstein-Barr virus lymphoproliferative disease (EBV-LPD) following allogeneic stem cell transplantation (allo-SCT) has a poor prognosis. We used a sensitive real-time polymerase chain reaction (PCR) assay for quantitative detection of EBV DNA in plasma and serially measured EBV-DNA levels to assess the response to treatment in allo-SCT recipients with EBV-LPD. Fourteen allo-SCT recipients with EBV-LPD who received a T cell-depleted (TCD) sibling (n = 5) or matched unrelated donor (n = 9) graft were monitored from the time of EBV-LPD diagnosis, during therapy and assessment of clinical response. Seven patients had complete responses of EBV-LPD to therapy, of whom 21% (3 out of 14) survived beyond 6 months from EBV-LPD diagnosis. Clinically responding patients showed a rapid decline of EBV-DNA plasma levels within 72 h from the start of therapy. In contrast, all clinical non-responders showed an increase of EBV-DNA levels. Absolute EBV-DNA levels at the time of EBV-LPD diagnosis did not predict for response, but the pattern of EBV-DNA levels within 72 h from the start of therapy (> 50% decrease versus increase) strongly predicted for clinical response (P = 0.001). Quantitative monitoring of EBV-DNA levels from the start of and during therapy for EBV-LPD rapidly and accurately predicts for response to therapy as early as within 72 h. It may thus provide a powerful tool to adjust and select treatment in individuals with EBV-LPD following allo-SCT. PMID- 11380476 TI - Effect of chemotherapy and stem cell transplantation on T lymphocyte clones in familial haemophagocytic lymphohistiocytosis. AB - Familial haemophagocytic lymphohistiocytosis (FHL) is a rare disorder in infancy, curative only by an allogeneic stem cell transplantation (SCT). We recently confirmed the clonal evidence of T cells in FHL. To confirm the effect of chemotherapy and SCT in FHL, the change of T-cell clones was analysed in two patients using inverse reverse transcription-polymerase chain reaction (RT-PCR) of the T-cell receptor variable region (TCR V) gene, followed by PCR for the junctional region (Jbeta-PCR), a single-strand conformation polymorphism (SSCP) and sequencing analysis at diagnosis, after chemotherapy and after SCT. A high frequency (> 15%) of alphabeta T-cell clones and a predominant bias (Jbeta1:Jbeta2, 85:15) for the Jbeta1 subgroup were observed in the two patients at diagnosis. In one patient, however, an inverted predominant bias (Jbeta1:Jbeta2, 9:91) for the Jbeta2 subgroup and oligoclonal expansion were observed at relapse after chemotherapy. In the other patient, correction of both restricted Jbeta cluster usage and variation of TCR were observed after chemotherapy and SCT. Using sequence analysis, the clonal T cells detected at diagnosis were found to be substituted at low frequency (< 0.75%) by several new clones after chemotherapy and SCT. These results indicate that any genetic defect could influence the regulation of the T-cell network, and normalization of both the variation in each Vbeta repertoire and the Jbeta1/Jbeta2 ratio is needed to achieve remission, and might support the rationale that the only acceptable curative therapy of FHL is allogeneic SCT. PMID- 11380477 TI - Changes in the expression of FGFR3 in patients with chronic myeloid leukaemia receiving transplants of allogeneic peripheral blood stem cells. AB - Fibroblast growth factor receptors (FGFR1-4) are implicated in various cellular events, including cell growth and transformation. Here, we showed that patients suffering from chronic myeloid leukaemia (CML) express high levels of FGFR3 mRNA in white blood cells (WBCs). After stem cell transplantation and reconstitution of haematopoiesis, the expression of FGFR3 decreased and was maintained at low levels that are typical of healthy individuals. However, FGFR3 expression became upregulated again in those patients that had accelerated BCR/ABL rearrangement and underwent relapse of leukaemia. Our findings suggest that, in CML, the changing levels of FGFR3 transcripts in WBCs may have prognostic significance. PMID- 11380478 TI - Non-transferrin-bound iron during allogeneic stem cell transplantation. AB - Hydroxyl radical formation catalysed by non-transferrin-bound iron (NTBI) might contribute to transplantation-related complications. The occurrence of NTBI in 10 adult allogeneic stem cell transplantation (SCT) patients was followed for 20 d. The transferrin saturation reached 99% on d -4 and remained > 80% thereafter. NTBI, measured as bleomycin-detectable iron, was detected for 6-18 d in all patients with a peak on d -4. High transferrin saturation levels were associated with the appearance of NTBI with a threshold at 80% saturation. Prevention of the potential deleterious effects of NTBI might reduce transplantation-related morbidity. PMID- 11380479 TI - The use of oral vitamin K for reversal of over-warfarinization. PMID- 11380480 TI - Autologous stem cell transplantation for rapidly progressive Jo-1-positive polymyositis with long-term follow-up. PMID- 11380481 TI - Bisphosphonates and in vivo models of multiple myeloma. PMID- 11380483 TI - Formaldehyde promotes and inhibits the proliferation of cultured tumour and endothelial cells. AB - Formaldehyde was applied in various doses (0.1-10.0 mM) to HT-29 human colon carcinoma and HUV-EC-C human endothelial cell cultures. Cell number, apoptotic and mitotic index as well as proportion of cells in S-phase was investigated by morphological methods and flow cytometry. Ten mM of formaldehyde caused high degree of cell damage and practically eradicated the cell cultures. One mM of formaldehyde enhanced apoptosis and reduced mitosis in both types of cell cultures, in a moderate manner. The low dose (0.1 mM) enhanced cell proliferation and decreased apoptotic activity of the cultured cells, the tumour cells appeared to be more sensitive. The possible role of this dose-dependent effect of formaldehyde in various pathological conditions, such as carcinogenesis and atherogenesis is discussed with emphasis on the eventual interaction between formaldehyde and hydrogen peroxide. PMID- 11380484 TI - Differential mitogenic responses of human macrovascular and microvascular endothelial cells to cytokines underline their phenotypic heterogeneity. AB - A variety of growth factors promote the complex multistep process of angiogenesis. The mitogenic activity of vascular endothelial growth factors (VEGFs) and placental growth factors (PlGFs), known as cytokines acting predominantly on endothelial cells, was tested on human umbilical vein endothelial cells (HUVEC) and microvascular endothelial cells (MIEC) and compared with the potency of the universally acting basic fibroblast growth factor (FGF 2). The cells were seeded at different cell numbers and incubated with various doses of growth factors for a period of 24-72 h in culture medium +/- serum. Proliferation was determined by measuring the optical density after staining the cells with the tetrazolium salt WST-1. VEGF121 and VEGF165 increased the number of HUVEC and MIEC at low and high seeding densities various doses and incubation times. The efficiency of FGF-2 was less pronounced at high seeding densities of the cells under serum-free conditions. PlGF-1 and PlGF-2 stimulated mitogenesis on HUVEC only at low cell numbers and after a short incubation time by 125 +/- 3% and 102 +/- 5% (P < 0.001), respectively. Longer incubation times with the lower seeding density in the absence of FCS did not induce a significant stimulatory effect of the PlGFs. MIEC responded stronger to all growth factors. In particular under serum free conditions, PlGF-1 and PlGF-2 effectively stimulated cell proliferation by 247 +/- 54% (P < 0.01) and 288 +/- 40% (P < 0.05) at low cell numbers, and by 81 +/- 13% (P < 0.05) and 49 +/- 13% (P < 0.01), respectively, at high cell numbers. The addition of fetal calf serum caused a reduced proliferative response of all growth factors on both cell types related to the controls. In conclusion, MIEC and HUVEC differ in their proliferative response to VEGFs, PlGFs and FGF-2. PMID- 11380485 TI - Aromatase regulation and breast cancer. PMID- 11380486 TI - Paraganglioma genes. PMID- 11380487 TI - Effects of the selective oestrogen receptor modulator-raloxifene-on calcium and PTH secretory dynamics in women with osteoporosis. AB - OBJECTIVES: A possible mechanism for the maintenance of bone mass by oestrogens and the selective oestrogen receptor modulator (SERM)-raloxifene-is an interaction with calciotropic hormones. We studied the effects of raloxifene on calcium-PTH homeostasis. PATIENTS AND MEASUREMENTS: Calcium and EDTA infusions were performed in 32 post-menopausal women with osteoporosis (BMD T score < - 2.5). This cross-sectional study was performed in the third year of the MORE (Multiple Outcomes of Raloxifene Evaluation) trial, a double-blind, placebo controlled study. After an overnight fast, calcium glubionate (5 mg/kg BW*h), and after 2.5 h of test-free interval, Na3EDTA (40 mg/kg BW*h) were given intravenously. The duration of infusions was based on individual plasma total calcium before the calcium infusion (t = 0), the target calcium (2.60 and 1.95 mmol/l, respectively), and desired mean calcium change (0.010 mmol/L*min). Blood samples were taken at 0 and every 5 minutes of both infusions. Plasma PTH levels were fitted into an inversed sigmoidal relation with plasma calcium. The effect of raloxifene on calcium-PTH homeostasis was tested in linear regression models adjusted for age and BMI. Nine patients used placebo, 13 raloxifene 60 mg/day and 10 raloxifene 120 mg/day. RESULTS: Raloxifene use was associated with lower plasma albumin (40.7 +/- 1.8 vs. 38.0 +/- 2.0 and 38.5 +/- 2.3 g/l, for placebo, raloxifene 60 mg/day and raloxifene 120 mg/day, respectively, P = 0.01), lower plasma total calcium at t = 0 (2.28 vs. 2.24 and 2.21; +/- 0.07 mmol/L; P = 0.03), lower plasma total calcium at 50% of maximal PTH secretion (PTH set-point: 2.23 +/- 0.06 vs. 2.18 +/- 0.07 and 2.16 +/- 0.08 mmol/l, P = 0.06), and lower plasma non-suppressible PTH (0.84 +/- 0.19 vs. 0.75 +/- 0.10 and 0.73 +/- 0.05 pmol/l, P = 0.02). After correction for plasma albumin, the differences for plasma calcium at t = 0 and at PTH set-point were no longer significant. In contrast, the difference in PTH suppression during calcium load was not explained either by differences in plasma albumin or calcium. CONCLUSION: Raloxifene did not have any detectable effect on the PTH set-point. An effect on non suppressible PTH secretion cannot be excluded. PMID- 11380488 TI - Relationships with serum parathyroid hormone in old institutionalized subjects. AB - OBJECTIVE AND BACKGROUND: Old people in residential care are at the highest risk of any group for hip fracture. This may relate to their high prevalence of hyperparathyroidism. There are few data, however, on relationships with serum parathyroid hormone (PTH) in these individuals. This study therefore examined complex associations with serum PTH in nursing home and hostel residents. DESIGN: Cross-sectional analysis. PATIENTS: One hundred and forty-three nursing home and hostel residents of median age 84 years. MEASUREMENTS: Serum PTH, 25 hydroxyvitamin D (25OHD), 1,25-dihydroxyvitamin D (1,25-(OH)2D), plasma creatinine, phosphate, calcium, albumin, Bsm-1 vitamin D receptor genotype, age, weight and use of frusemide or thiazide. RESULTS: The statistical models determined accounted for half the interindividual variation in serum PTH. Heavier weight was associated with both the prevalence of secondary hyperparathyroidism and the serum concentration of PTH. Novel interactions with serum PTH were identified between: weight and 25OHD; 25OHD and phosphate; and phosphate and thiazide diuretic use. Plasma phosphate was associated with PTH independently of calcium and 1,25-(OH)2D. There was no independent association between PTH and nuclear vitamin D receptor genotype. CONCLUSIONS: Heavier weight is associated with both the prevalence and severity of secondary hyperparathyroidism and consistent with animal models of secondary hyperparathyroidism, phosphate may relate to serum PTH independently of 1,25-(OH)2D or calcium. PMID- 11380489 TI - Prevalence of diabetes and impaired glucose tolerance in adult hypopituitarism on low dose oral hydrocortisone replacement therapy. AB - OBJECTIVE: The conventional dosage of hydrocortisone, used for many years in the management of hypopituitarism (30 mg per day), has now been shown to be more than is physiologically necessary. On this conventional corticosteroid therapy studies have demonstrated an increased prevalence of diabetes and impaired glucose tolerance, which may contribute to the increased vascular morbidity and mortality reported in the condition. In these studies no information is available on oral glucose tolerance test (OGTT) timing in relation to administration of oral steroid and variable hydrocortisone doses were employed. PATIENTS: In order to assess glucose tolerance in patients treated with lower, more physiological doses, we performed a 75-g OGTT at least 1 month after hydrocortisone therapy was adjusted to 15 mg at 0800 h and 5 mg at 1700 h in 45 adult onset hypopituitary patients (30 M, 15 F). Mean (+/- SD) duration of hypopituitarism was 12 +/- 10 years, mean age 52 +/- 14 years and BMI 29.3 +/- 5.1 kg/m2. All were on hydrocortisone, 43 on thyroxine, 31 on sex steroids, 9 on desmopressin and 33 had documented growth hormone deficiency. Hydrocortisone 15 mg was taken at 0800 and the OGTT commenced at 0900. RESULTS: Using standard WHO criteria 36 patients (80%) had normal glucose tolerance, 1 (2%) had newly diagnosed diabetes and 8 (18%) had impaired glucose tolerance. Using the recently announced American Diabetes Association criteria for diagnosis 96% had normal glucose tolerance, 2% had diabetes and 2% impaired fasting glucose. CONCLUSION: The markedly reduced prevalence of diabetes and impaired glucose tolerance on lower hydrocortisone replacement doses in our series of patients with hypopituitarism, not previously known to be diabetic, is of great interest. This lower prevalence may eventually result in reduced vascular complication rates. PMID- 11380490 TI - The corticotropin-releasing hormone test in the diagnosis of ACTH-dependent Cushing's syndrome: a reappraisal. AB - OBJECTIVE: Stimulation with corticotropin-releasing hormone (CRH) is one of principal tools for the differential diagnosis of ACTH-dependent Cushing's syndrome. However, different dosages and species of CRH may be employed; further, ACTH levels can be measured by radioimmunoassay (RIA) or immunoradiometric assay (IRMA). The aims of the present study were to perform a reappraisal of the diagnostic accuracy of the CRH test taking these different testing modalities into consideration and to study the correlation between basal ACTH and cortisol levels and their responses to CRH in patients with Cushing's disease. PATIENTS: The study population comprised 148 patients with Cushing's disease and 12 patients with ectopic ACTH secretion collected through an Italian multicentre study. DESIGN: Patients were submitted to stimulation with 100 microg human or ovine CRH (36% and 64% of subjects, respectively) and ACTH measured either by RIA or IRMA (28% and 72%, respectively). A 50% increase in ACTH and cortisol levels was considered indicative of Cushing's disease. RESULTS: Mean peak ACTH levels measured by RIA and IRMA were comparable, as was the diagnostic accuracy of the test with the two assays (87% for IRMA and 84% for RIA, ns). In patients with Cushing's disease, stimulation with ovine CRH induced greater hormonal responses compared to testing with human CRH although only the cortisol response reached statistical significance (ACTH: 247.5 +/- 28.0% vs. 168.5 +/- 21.3% over baseline, P = 0.06; cortisol: 89.3 +/- 8.5% vs. 60.8 +/- 9.6% over baseline, P < 0.05 for ovine and human CRH, respectively). No appreciable rise in ACTH and cortisol levels was registered among patients with ectopic ACTH secretion. Diagnostic accuracy of the cortisol response was significantly greater with the ovine than with human peptide (71% vs. 49%, P < 0.05, for ovine and human CRH, respectively) while the ACTH response yielded equal diagnostic accuracy (86% vs. 87%, ns, for the ovine and human peptide, respectively). Interestingly, the correlation between ACTH and cortisol peak responses in patients with Cushing's disease was significantly greater for human than for ovine CRH (r = 0.68 vs. r = 0.41, P < 0.01, respectively). In addition, baseline cortisol levels exhibited a significant negative correlation with both the ACTH and cortisol response to CRH suggesting the persistence of the negative cortisol feedback in patients with Cushing's disease. CONCLUSIONS: (A) Both RIA and IRMA can be used indifferently for the assessment of the ACTH response to CRH. (B) Human and ovine CRH provide the same diagnostic accuracy as regards the ACTH response which, incidentally, represents the most accurate criterion for the evaluation of the CRH test; ovine CRH is superior to the human peptide in the evaluation of the cortisol response. (C) In patients with Cushing's disease, endogenous cortisol maintains the ability to negatively modulate CRH-stimulated corticotropin secretion. PMID- 11380491 TI - The limited value of the desmopressin test in the diagnostic approach to Cushing's syndrome. AB - OBJECTIVE: The desmopressin test is generally regarded as an alternative to the CRH test but it is unclear whether desmopressin is as effective as CRH in the differential diagnosis of ACTH-dependent Cushing's syndrome. However, a precise assessment of the operating characteristics of the desmopressin test in comparison with the CRH test has not been reported. The aim of the present study was to make a comparative evaluation of desmopressin and CRH tests in a consecutive cohort of patients with ACTH-dependent Cushing's syndrome and in a group of healthy subjects. DESIGN AND SUBJECTS: We studied 34 patients with Cushing's disease (CD) and nine patients with ectopic ACTH syndrome (EAS). The control group included 30 healthy subjects. Estimates of sensitivity and specificity were determined for a value of ACTH percent increment (Delta%) > 35% and for a Delta % > 50%, following either desmopressin or CRH, to differentiate CD from EAS. The sensitivity and specificity of a composite rule requiring an ACTH net increment (Delta) > 4.5 pmol/l at both values of Delta % was also calculated. When evaluating cortisol responses, the criteria were Delta % > 20% and Delta > 193 nmol/l. Moreover, to allow comparison of individual end points of the desmopressin and CRH tests at multiple levels of Delta % or Delta either for ACTH or cortisol without the bias of predetermined criteria, univariate curves of the receiver operating characteristics (ROC) were constructed by plotting the sensitivity against 1 - specificity at each level. RESULTS: In the patients with CD, the frequency of ACTH response was of 90% after both tests while the figures for cortisol were 73% after CRH and 77% after desmopressin, respectively. In the 15 patients who underwent both tests the magnitude of ACTH and cortisol responses induced by the 2 stimuli were fully comparable. In the patients with EAS a (false) positive ACTH response was found in 2/9 cases (22%) after the CRH test and in 2/5 patients (40%) after the desmopressin test. In the healthy subjects the CRH test was performed in 25 cases and the desmopressin test in 15 cases. The frequency of ACTH response was 52% following CRH and 13% following desmopressin. In the 10 healthy subjects who underwent both tests the ACTH response was significantly greater after CRH than desmopressin. The area under the ROC curve for the ACTH Delta % was significantly different than that occurring by chance following CRH but not desmopressin. The point on the ROC curve closest to 1 corresponded to an ACTH Delta % of 47% (sensitivity 87% and specificity 89%). However, a criterion of 100 % specificity would require an increase in the threshold for the ACTH Delta % to 259%. ROC analysis validated also the use of the ACTH Delta as a method to assess the response to CRH, but not after desmopressin. However, the diagnostic performance of this parameter was reduced in comparison to that of the ACTH Delta %, since the best cut-off for the Delta (6.2 pmol/l) had inferior specificity (79%). The operating characteristics of CRH and desmopressin were worse when considering cortisol responses. CONCLUSIONS: The present data suggest that the CRH test is more reliable than the desmopressin test in determining the aetiology of Cushing's syndrome. The desmopressin test resulted in a high frequency of false positive results in patients with ectopic ACTH secondary to carcinoid tumours. This finding may be due to the capability of these tumours to express the V3 vasopressin receptor through which desmopressin acts. However, the clinical endocrinologist may be confronted with some disturbing cases which are misdiagnosed because it is almost impossible to set a diagnostic criterion providing complete specificity in the differentiation of (occult) ectopic Cushing's syndrome using either CRH or desmopressin tests. PMID- 11380492 TI - Transsphenoidal pituitary surgery in Cushing's disease: can we predict outcome? AB - OBJECTIVE: To assess the results of transsphenoidal pituitary surgery in patients with Cushing's disease over a period of 18 years, and to determine if there are factors which will predict the outcome. PATIENTS: Sixty-nine sequential patients treated surgically by a single surgeon in Newcastle upon Tyne between 1980 and 1997 were identified and data from 61 of these have been analysed. DESIGN: Retrospective analysis of outcome measures. MAIN OUTCOME MEASURES: Patients were divided into three groups (remission, failure and relapse) depending on the late outcome of their treatment as determined at the time of analysis, i.e. 88 months (median) years after surgery. Remission is defined as biochemical reversal of hypercortisolism with re-emergence of diurnal circadian rhythm, resolution of clinical features and adequate suppression on low-dose dexamethasone testing. Failure is defined as the absence of any of these features. Relapse is defined as the re-emergence of Cushing's disease more than one year after operation. Clinical features such as weight, sex, hypertension, associated endocrine disorders and smoking, biochemical studies including preoperative and postoperative serum cortisol, urine free cortisol, serum ACTH, radiological, histological and surgical findings were assessed in relation to these three groups to determine whether any factors could reliably predict failure or relapse after treatment. RESULTS: Of the 61 patients included in this study, 48 (78.7%) achieved initial remission and 13 (21.3%) failed treatment. Seven patients suffered subsequent relapse (range 22-158 months) in their condition after apparent remission, leaving a final group of 41 patients (67.2%) in the remission group. Tumour was identified at surgery in 52 patients, of whom 38 achieved remission. In comparison, only 3 of 9 patients in whom no tumour was identified achieved remission. This difference was significant (P = 0.048). When both radiological and histological findings were positive, the likelihood of achieving remission was significantly higher than if both modalities were negative (P = 0.038). There were significant differences between remission and failure groups when 2- and 6-week postoperative serum cortisol levels (P = 0.002 and 0.001, respectively) and 6-week postoperative urine free cortisol levels (P = 0.026) were compared. This allowed identification of patients who failed surgical treatment in the early postoperative period. Complications of surgery included transitory DI in 13, transitory CSF leak in 8 and transitory nasal discharge and cacosmia in 3. Twelve of 41 patients required some form of hormonal replacement therapy despite achieving long-term remission. Thirteen patients underwent a second operation, of whom 5 achieved remission. CONCLUSIONS: Transsphenoidal pituitary surgery is a safe method of treatment in patients with Cushing's disease. Operative findings, radiological and histological findings, together with early postoperative serum cortisol and urine free cortisol estimates may identify failures in treatment. Alternative treatment might then be required for these patients. Because of the risk of late relapse, patients require life-long follow-up. PMID- 11380493 TI - Angiotensin II type 1 receptor and ACTH receptor expression in human adrenocortical neoplasms. AB - OBJECTIVE: Type 1 angiotensin II (Ang II) receptors transduce most of the known actions of Ang II, including steroidogenesis and trophic actions on the adrenal cortex. We investigated the type 1 Ang II receptor expression in adrenocortical tissues to define its regulation in adrenocortical neoplasms and to compare its expression with that of the ACTH receptor (ACTH-R). PATIENTS AND MEASUREMENTS: Poly A RNA was extracted from tumour tissue and electrophoresed through a 1.0% agarose gel, blotted and hybridized with alpha32P-CTP labelled PCR generated type 1 Ang II receptor cDNA probe. Receptor autoradiography was performed on slices from normal adrenals and tumour tissue by incubation with 125I-Sar1, Ile8-Ang II with and without pretreatment with cold Ang II or with the selective type 1 receptor antagonist losartan. RESULTS: Ang II type 1 receptor mRNA was high in cortisol producing (CPA; n = 5) and aldosterone producing (APA; n = 4) adenomas (normal adrenals 100 +/- 12% vs. 180 +/- 16% in CPA and 154 +/- 26% in APA, mean +/- SEM), but was low in nonfunctioning adenomas (NFA; n = 2; 2 +/- 1%). ACTH receptor mRNA followed a similar pattern (CPA 178 +/- 17, APA 196 +/- 30, NFA 0%, carcinomas 56 +/- 11%) with a good correlation between Ang II type 1 receptor and ACTH-R mRNA of r = 0.692, P = 0.0019. Receptor autoradiography in normal adrenals demonstrated Ang II type 1 receptors predominantly in the zona glomerulosa. In tumour tissue, mainly type 1 receptor expression was found confirming the Northern blot data. CONCLUSIONS: Angiotensin II type 1 receptor and ACTH receptor expression seems to be correlated with the functional status of adrenocortical tumours, suggesting regulation by similar factors. The predominant receptor expressed in adrenocortical tumours is the Angiotensin II type 1 receptor whereas type 2 receptor expression is minimal. PMID- 11380494 TI - A critical examination of adrenal tuberculosis and a 28-year autopsy experience of active tuberculosis. AB - OBJECTIVE: Tuberculosis is potentially fatal and adrenal gland involvement is uncommonly reported. The aims of the current study were to define the characteristics of tuberculosis in hospitalized patients and to analyse the features of adrenal tuberculosis. DESIGN: Retrospective analysis of autopsies and adrenalectomies. PATIENTS: 13,762 patients (13492 at autopsies and 270 at adrenalectomy). MEASUREMENTS: The presence of active tuberculosis, the predisposing factors, the pathological features and organs of involvement were examined. RESULTS: Active tuberculosis was present in 871 patients (6.5% of all 13492 autopsies). It was first diagnosed in 70% of these patients during autopsy. Cancers and a history of recent major operations were the 2 main concomitant factors in the patients with tuberculosis. Extra-pulmonary tuberculosis was seen in 261 patients (30%). The five most common extra-pulmonary sites of tuberculosis were the liver, spleen, kidney, bone and adrenal gland. Adrenal tuberculosis was seen in 52 of the 871 patients (6%) with active tuberculosis at autopsy and in 3 patients at adrenalectomy. The adrenal gland was the only organ involved by active tuberculosis in 14 of these 55 patients (25%; 35 men, 20 women). Tuberculosis was evident on macroscopic examination of the adrenal glands in 46% of the patients. On histological examination, caseous necrosis and granulomatous inflammation with Langhan's giant cells were seen in 71% and 40% of patients, respectively. Seven patients presented with signs and symptoms of Addison's disease due to bilateral adrenal involvement. Langhan's giant cells were frequently seen in histological sections and bilateral enlargement of the adrenal glands was often noted. Fine needle aspiration cytology was not useful for diagnosing adrenal tuberculosis. CONCLUSION: Unexpected and extra-pulmonary tuberculosis such as adrenal tuberculosis has been a common problem. A high index of suspicion, correct diagnosis and proper treatment are essential for the management of tuberculosis. PMID- 11380495 TI - Immunohistochemical detection of somatostatin receptor types 1-5 in medullary carcinoma of the thyroid. AB - BACKGROUND: We have analysed the distribution of the five somatostatin receptors (sst1-5) by immunohistochemistry in a large retrospective series of 51 medullary carcinoma of the thyroid (MCT) specimens and correlated the pattern of sst expression with expression of somatostatin (SRIF) peptide, tumour pathology and clinical outcome. MEASUREMENTS: Immunohistochemistry was performed with rabbit polyclonal antipeptide antibodies directed against the extracellular domains or cytoplasmic tail of human (h) sst1-5. SRIF immunoreactivity was investigated in parallel paraffin sections. RESULTS: Eighty-five percent of the tumours were positive for one or more sst, localized to both tumour cells as well as surrounding peritumoural structures, especially blood vessels. Forty-nine percent of the tumours were positive for sst1, 43% for sst2, 47% for sst3, 4% for sst4, and 57% for sst5. Fifty-one percent of tumours expressed one or two sst subtypes; 33% were positive for three or more sst isoforms. All five sst receptors were detected in only two cases. Tumours expressing octreotide sensitive subtypes (sst2,3,5) accounted for 75% of the series. 50% of the tumours co-expressed SRIF suggesting tumour cell regulation by endogenous SRIF via paracrine/autocrine circuits. There was no correlation between sst1-5 expression and age, sex, tumour size or stage, histological type or clinical outcome. Simultaneous analysis of primary tumour and lymph node metastases revealed a similar pattern of sst immunoreactivity indicating that sst expression is not modified in the course of disease progression. CONCLUSIONS: With the exception of sst4, medullary carcinoma of the thyroid display a rich but heterogeneous expression of sst subtypes. Immunohistochemical typing of sst receptor expression using specific antireceptor antibodies represents an ideal approach for characterizing sst subtype expression in medullary carcinoma of the thyroid for optimizing receptor targeted diagnosis and therapy with somatostatin analogs. PMID- 11380496 TI - Frequency of appearance of myeloperoxidase-antineutrophil cytoplasmic antibody (MPO-ANCA) in Graves' disease patients treated with propylthiouracil and the relationship between MPO-ANCA and clinical manifestations. AB - OBJECTIVE: Myeloperoxidase antineutrophil cytoplasmic antibody (MPO-ANCA) positive vasculitis has been reported in patients with Graves' disease who were treated with propylthiouracil (PTU). The appearance of MPO-ANCA in these cases was suspected of being related to PTU because the titres of MPO-ANCA decreased when PTU was stopped. Nevertheless, there have been no studies on the temporal relationship between the appearance of MPO-ANCA and vasculitis during PTU therapy, or on the incidence of MPO-ANCA in untreated Graves' disease patients. Therefore, we sought to address these parameters in patients with Graves' disease. PATIENTS: We investigated 102 untreated patients with hyperthyroidism due to Graves' disease for the presence of MPO-ANCA, and for the development vasculitis after starting PTU therapy. Twenty-nine of them were later excluded because of adverse effects of PTU or because the observation period was less than 3 months. The remaining 73 patients (55 women and 18 men), all of whom were examined for more than 3 months, were adopted as the subjects of the investigation. The median observation period was 23.6 months (range: 3-37 months). MEASUREMENTS: MPO-ANCA was measured at intervals of 2-6 months. RESULTS: Before treatment, the MPO-ANCA titres of all 102 untreated Graves' disease patients were within the reference range (below 10 U/ml). Three (4.1%) of the 73 patients were positive for MPO-ANCA at 13, 16 and 17 months, respectively, after the start of PTU therapy. In two of them, the MPO-ANCA titres transiently increased to 12.8 and 15.0 U/ml, respectively, despite continued PTU therapy, but no vasculitic disorders developed. In the third patient, the MPO-ANCA titre increased to 204 U/ml and she developed a higher fever, oral ulcers and polyarthralgia, but the symptoms resolved 2 weeks after stopping PTU therapy, and the MPO-ANCA titre decreased to 20.7 U/ml by 4 months after discontinuing PTU. CONCLUSIONS: PTU therapy may be related to the appearance of MPO-ANCA, but MPO ANCA does not appear to be closely related to vasculitis. PMID- 11380497 TI - Plasma levels of insulin-like growth factor binding protein-4 (IGFBP-4) under normal and pathological conditions. AB - OBJECTIVE: Insulin-like growth factor binding protein-4 (IGFBP-4) belongs to a family of six structurally related IGF-binding proteins that are involved in the modulation of the biological effects of the IGFs. In order to obtain more insight into the clinical significance and regulation of IGFBP-4 in vivo we determined the levels of this protein by a specific radioimmunoassay in the human circulation under normal and various pathological conditions. DESIGN AND PATIENTS: Selected human biological fluids and plasma samples from 804 normal healthy males and females, ranging from 0 to 78 years of age, were analysed. In addition, plasma samples from patients with several disorders (i.e. hypothyroidism, hyperthyroidism, GH-deficiency, acromegaly, cancer, chronic renal failure corticosteroid-treatment) were investigated. MEASUREMENTS: A specific RIA for hIGFBP-4 was developed, using a rabbit polyclonal antibody raised against a synthetic peptide containing amino acids 80-103 of the mature hIGFBP-4 sequence. RESULTS: In normal individuals circulating IGFBP-4 levels in males did not change with age. For females the values tended to increase slightly in older age. Overall, the mean +/- SD for males and females (189 +/- 83 microg/l and 193 +/- 72 microg/l, respectively) were not different. Normative range values of IGFBP-4 correlated weakly with those of IGF-II (r = 0.31, P < 0.001). Neither hypothyroidism nor hyperthyroidism appeared to influence circulating IGFBP-4 levels since the levels were within the normal range. Both GH status and pharmacological doses of glucocorticoids, as employed in various chronic diseases, did not seriously affect plasma IGFBP-4 either. Under conditions with increased circulating PTH levels, i.e. dialysed adult patients with chronic renal failure (CRF) and subjects with hyperparathyroidism, a weak positive relationship was noted between the plasma contents of IGFBP-4 and PTH. An excess of IGFBP-4 was found in plasma of both nondialysed and dialysed prepubertal growth retarded children with chronic renal failure (CRF) (mean SDS: 10.75 and 5.78, respectively). IGFBP-4 levels were inversely related to glomerular filtration rate. Similar results were obtained for dialysed adults with CRF. In a group of CRF children who had undergone renal transplantation, circulating IGFBP-4 levels were markedly lower (mean SDS: 3.75). There was no evidence for an increased secretion of IGFBP-4 in the circulation of most of the cancer patients with solid tumours. Several children with acute lymphoblastic leukaemia, however, showed elevated plasma IGFBP-4 levels (mean SDS: 1.27). The presence of IGFBP-4 could also be demonstrated in other human biological fluids. The highest amounts were found in amniotic fluid (391-717 microg/l) and follicular fluid (249-500 microg/l). CONCLUSIONS: Measurement of plasma IGFBP-4 has been shown so far to be of minor clinical relevance. However, the results indicate that different concentration gradients between plasma and various other body fluids may exist. Therefore, it may well be that certain pathophysiological stimuli induce significant alterations in the local turnover rate of IGFBP-4 but that they are not reflected by changes in the circulating levels. The possibility of quantifying IGFBP-4 by RIA will facilitate further in vitro and in vivo studies on its regulation and function in humans. PMID- 11380498 TI - Differential diagnosis of polyuric/polydipsic syndromes with the aid of urinary vasopressin measurement in adults. AB - OBJECTIVE: A water deprivation test or a hypertonic saline infusion test with the measurement of plasma osmolality and plasma vasopressin are the gold standard tests in the differential diagnosis of polyuric syndromes. Because commercially available vasopressin kits are too insensitive for this approach, and the concentration of vasopressin in urine is much higher than in plasma, urinary vasopressin measurements may be an alternative to the more difficult plasma vasopressin measurement. DESIGN: The diagnostic value of the measurement of urinary vasopressin with a rather insensitive commercially available vasopressin kit was compared with plasma vasopressin measurement by a highly sensitive radioimmunoassay (RIA). PATIENTS AND METHODS: Thirteen normal subjects and 27 patients with polyuria/polydipsia were examined by an 8-h fluid deprivation test. In all blood samples (0800 h, 1200 h, 1400 h and 1600 h) and in all urine collections (2-hourly fractions), osmolality as well as vasopressin were measured. RESULTS: Using plasma vasopressin measurement with a highly sensitive RIA as gold standard test, nine patients were classified as having primary polydipsia, whereas 18 had partial or complete cranial diabetes insipidus. Whereas the substitution of plasma vasopressin measurement by urinary vasopressin measurement alone did not provide 100% separation between both groups, the product of urinary vasopressin and urinary osmolality related to plasma osmolality completely separated the patients with primary polydipsia from those with diabetes insipidus. Urinary measurement of vasopressin and osmolality alone, which was recommended as a noninvasive diagnostic procedure in children, was too insensitive for exact differential diagnosis in our adult patients. CONCLUSIONS: The simultaneous measurement of plasma vasopressin and plasma osmolality in a dehydration test is the most powerful diagnostic tool in the differential diagnosis of polyuria/polydipsia. However, if highly sensitive assays for plasma vasopressin measurements are not available, the measurement of urinary vasopressin with commercially available, less sensitive RIAs may be a diagnostic alternative, which showed nearly the same sensitivity as plasma vasopressin measurement in our study population. PMID- 11380499 TI - Changes of leptin and metabolic hormones in preterm infants: a longitudinal study in early postnatal life. AB - OBJECTIVES: Very little is known concerning the physiological role of leptin and growth in the early postnatal period and the association of leptin with other metabolic hormones in preterm infants. This study aims to investigate these relationships, and to explore the longitudinal and dynamic profile of leptin and metabolic hormones including insulin, ACTH, cortisol and FT4 in this category of patient. We also postulate that a rapid increase in body weight and body mass index in the first few weeks of life may be associated with a corresponding increase in serum leptin if the 'adipoinsular axis' is active at this stage. DESIGN: A longitudinal study in a cohort of preterm infants < 34 weeks gestation for the first five weeks of postnatal life. PATIENTS AND METHODS: Sixty-one preterm newborns < 34 weeks gestation were prospectively enrolled. Blood samples were collected in the morning when the newborns were 24 h of age (day 1), and on days 5, 14 and 35 of life. Serum leptin, insulin, cortisol, FT4, glucose and plasma ACTH concentrations were analysed using standard biochemical methods. Spearman's correlation coefficient was used to assess the inter-relationship of different metabolic hormones on the first day of life, and the relationship between metabolic hormones on day 1 and anthropometric or clinical parameters. The mixed-effects models were further used for analysing the multiple longitudinal measurements, and also for comparing the hormone concentrations between day 1 (baseline) and their corresponding levels on days 5, 14 and 35. RESULTS: Serum leptin on day 1 (baseline) was significantly associated with serum insulin (r = 0.30, P < 0.05) and insulin:glucose ratio (r = 0.29, P < 0.05) in infants < 34 weeks gestation. Female preterm infants had significantly higher serum leptin (P < 0.05) and insulin (P < 0.05) levels than male infants. In addition, the duration between the last dose of antenatal dexamethasone and delivery significantly influenced the serum concentrations of leptin (r = - 0.27, P < 0.05), cortisol (r = 0.52, P < 0.001), plasma ACTH (r = 0.28, P < 0.05) and insulin:glucose ratio (r = - 0.27, P < 0.05) on the first day of life. Despite significant increase in body weight (P < 0.00001) and body mass index (P < 0.00001) by day 35 when compared to birth weight, no corresponding significant changes were observed for serum leptin, insulin and FT4. However, there was an increasing though statistically nonsignificant trend in serum leptin after day 14 of life. CONCLUSION: This study characterized the longitudinal profile of leptin and metabolic hormones in preterm infants < 34 weeks gestation in the first 5 weeks of life. Serum leptin was significantly associated with serum insulin and insulin:glucose ratio supporting the hypothesis that an 'adipoinsular axis' exists and is likely to be functional before 34 weeks of gestation. Although a significant increase in body weight was demonstrated by day 35, no significant corresponding changes occurred with regard to serum leptin. We postulate that the limited quantity of adipose tissue at this stage of development might have contributed to this observation. Moreover, our results also showed that the duration between the last dose of antenatal corticosteroid and delivery could influence the postnatal concentrations of adipoinsular and pituitary-adrenal hormones. Thus, it is possible that antenatal dexamethasone might affect fetal growth and development via these neuroendocrine pathways in early intrauterine life. PMID- 11380500 TI - Isolated growth hormone (GH) deficiency due to compound heterozygosity for two new mutations in the GH-releasing hormone receptor gene. AB - OBJECTIVE: Mutations in the GH releasing hormone receptor (GHRH-R) have recently been shown to cause autosomal recessive isolated GH deficiency (IGHD). Patients who are homozygous for GHRH-R mutations have a subnormal GH response to pharmacological agents that stimulate GH secretion and an appropriate response to exogenous GH therapy. We searched for mutations in the GHRH-R gene in a family in which two of three siblings were affected by IGHD. DESIGN: We sequenced the 13 coding exons, the intron-exon boundaries and 327 bases of the promoter of the GHRH-R gene from peripheral blood cell genomic DNA of an index patient. RESULTS: Both affected individuals were compound heterozygotes for two previously undescribed GHRH-R mutations: a change in codon 137 that replaces histidine with leucine (H137L), and a 5 bp deletion in exon 11 (Del 1140-1144). The patients' father was heterozygous for the H137L mutation, and the mother was heterozygous for the exon 11 deletion. We used site-directed mutagenesis to create the mutants in wild-type GHRH-R cDNA. Transient transfection of GHRH-R cDNAs in Chinese hamster ovary cells showed that cells transfected with both mutant receptors failed to increase cyclic AMP after treatment with GHRH. CONCLUSIONS: We describe a family in which two siblings with IGHD were compound heterozygotes for two new mutations in the GHRH-R gene. These results suggest that mutant alleles for GHRH R gene may be more common than previously suspected. PMID- 11380501 TI - Cardiac phaeochromocytoma presenting with severe hypertension and chest pain. AB - Cardiac phaeochromocytoma is a rare cause of endocrine hypertension. We report a case of a 25-year-old woman, who presented with severe hypertension and intermittent chest pain. The patient denied typical phaeochromocytoma spells of palpitation, headache, and diaphoresis. The 24-hr urinary excretion of norepinephrine was increased sevenfold above the upper limit of normal; however, the excretion of total metanephrines, epinephrine, and dopamine were normal. Computed tomography (CT) scan of the abdomen was normal. An 131I-labelled metaiodobenzylguanidine (MIBG) scan was falsely negative while the patient was taking labetalol. The cardiac phaeochromocytoma was localized with indium-111 pentetreotide scintigraphy and chest magnetic resonance imaging scan. Repeat 123I MIBG scintigraphy was positive after discontinuing labetalol. The cardiac phaeochromocytoma was located in the right atrial groove, adjacent to the tricuspid valve, and contained multiple feeder arteries from the right coronary artery. After treatment with volume expansion, alpha-methyl-p-tyrosine, and alpha and beta-adrenergic blockade, surgical resection was performed. While under cardiopulmonary bypass, coronary bypass grafting and tricuspid annuloplasty were performed to facilitate the complete surgical resection of the 4.5-cm tumour. The surgical course was uncomplicated, with complete cure of hypertension and normalization of catecholamine excretion. Post-operative cardiac function, as measured by echocardiogram, was normal. Although cardiac phaeochromocytoma may be highly vascular, invasive and difficult to resect, it can be cured. PMID- 11380502 TI - Loss of gender difference in serum leptin levels and its slow recovery after successful surgery for Leydig cell tumours in two virilized females. AB - One of the factors that predicts serum leptin levels is gender. It has been shown that sex steroid hormones, in particular testosterone, play an important role in the regulation of serum leptin levels. We had the opportunity to examine the effects of acute and chronic changes in serum testosterone levels on serum leptin concentrations in two virilized females harbouring testosterone-secreting ovarian tumours, before and after curative surgery. Chronically elevated basal testosterone levels (46 nmol/l) were associated with suppressed serum leptin levels (1.46 microg/l and 2.56 microg/l) vs. 12 age- and BMI-matched healthy subjects 9.89 +/- 0.64 microg/l. Leptin levels were determined from pooled serum samples assayed by commercial radioimmunoassay. High testosterone levels abolished the well known sexual dimorphism of serum leptin levels. Two weeks after curative resection of these tumours serum leptin levels were unaltered and started to increase progressively after one month. One patient received parenteral conjugated oestrogens while the other resumed spontaneous menstrual cycles. Three months after curative surgery obvious changes in body composition were registered (DEXA). Six months later further rise in serum leptin concentrations occurred without further changes in body composition. In conclusion, leptin levels did not change in spite of rapid changes in the steroid milieu, but in the long term increase in body fat stores, new steroid milieu and maybe other factors are important determining factors of serum leptin levels. PMID- 11380503 TI - Diane 35 and spironolactone combination in the treatment of hirsutism. PMID- 11380505 TI - Cortisol responses to low (1 microg) and standard (250 microg) dose ACTH stimulation tests in patients with primary hypothyroidism. PMID- 11380506 TI - Dynamic aspects of platelet adhesion under flow. AB - 1. Cell-cell and cell-matrix adhesive interactions are critical for a wide range of physiological processes, including embryogenesis, inflammation, immunity and haemostasis. 2. The ability of circulating blood cells, such as platelets and leucocytes, to adhere to sites of vascular injury is complicated by the presence of blood flow, which imposes hydrodynamic forces on adhesion contacts. 3. To overcome this problem, platelets and leucocytes have evolved specific adhesion receptors with unique biomechanical properties that enable these cells to adhere to the vessel wall under flow conditions. 4. Platelet adhesion in the normal circulation appears to be a multiple-step process involving an initial reversible interaction between the platelet adhesion receptor glycoprotein Ib-IX-V and the vascular adhesion protein von Willebrand factor. Once tethered to the vessel wall, platelets form irreversible adhesion contacts through the binding of one or more platelet integrins to specific subendothelial matrix proteins. 5. There is now a wealth of evidence demonstrating that these receptors not only mediate platelet adhesion, but also transduce signals leading to platelet activation. 6. In the present review, we will briefly discuss the current understanding of the specific roles of individual platelet receptors in supporting the haemostatic function of platelets and discuss mechanisms by which these receptors induce platelet activation. PMID- 11380507 TI - Cardiac output in mice overexpressing beta2-adrenoceptors or with myocardial infarct. AB - 1. The aims of the present study were to characterize cardiac output (CO) in transgenic mice that overexpress the beta2-adrenoceptor and to evaluate ultrasonic flowmetery for continuous CO measurement in the mouse in vivo. 2. Under conditions of anaesthesia, open chest and positive ventilation, CO was determined with a transonic flowmeter at baseline and during dobutamine administration and intravenous volume loading in wild-type mice (n = 17) and beta2-adrenoceptor transgenic (n = 9) and wild-type mice with chronic myocardial infarct (n = 16). 3. Compared with wild-type mice, beta2-adrenoceptor transgenic mice with markedly enhanced ventricular contractility had a significantly higher CO, heart rate (HR) and maximal acceleration of aortic flow. Both dobutamine and volume loading increased CO in the two groups and higher levels of CO were measured in transgenic mice during the interventions. At baseline or during interventions, stroke volume was similar between beta2-adrenoceptor transgenic and wild-type mice. Infarcted mice with impaired cardiac function had a significantly lower CO under basal and stress conditions. 4. Thus, beta2 adrenoceptor transgenic mice revealed higher CO that was largely attributable to a significantly higher HR but not to an increase in stroke volume. Transonic flowmetery can detect differences in CO among mice in various functional states and is suitable for evaluation of cardiac functional reserve in mice in vivo by continuous monitoring of CO responses to different interventions. PMID- 11380508 TI - Endothelium-dependent changes in arterial diameter in old normotensive rats. AB - 1. In normotensive rats, removal of carotid artery endothelium results in an acute increase in diameter. This finding, observed in young animals, has not been investigated in old animals. The present study was undertaken to assess the contribution of endothelial function in the regulation of arterial stiffness in aged rats. 2. In normotensive female WAG/Rij rats, isobaric (100 mmHg transmural pressure) carotid diameter was measured in vitro in situ, using a previously described arterial preparation associated with a high-resolution echotracking technique allowing non-invasive diameter measurements under baseline conditions, after removal of the endothelium and after total relaxation of vascular smooth muscle by potassium cyanide. Histomorphometry of the carotid wall was studied after pressure fixation (100 mmHg) of the arteries. 3. Compared with younger animals (10 months), older animals (30 months) had the same baseline carotid isobaric diameter but significantly higher values of wall thickness and collagen content. In older animals, whereas total relaxation by potassium cyanide was associated with a slight but significant increase of isobaric diameter, no increase was observed after endothelium removal. 4. The results of the presnt study provide evidence that, in old normotensive rats, endothelium-dependent increases in isobaric carotid diameter are blunted. This endothelium alteration may contribute to the age-dependent increase in isobaric carotid stiffness observed in old rats. PMID- 11380509 TI - Effects in humans of intravenously administered endotoxin on soluble cell adhesion molecule and inflammatory markers: a model of human diseases. AB - 1. Endotoxin, a component of the cell wall of Gram-negative bacteria, could be a predisposing mediator of many pathological disorders. The present study was undertaken to determine the effects and time-course of acute endotoxin challenge on inflammatory and cell-adhesion molecule markers shedding in the plasma as potential surrogates. 2. Six normal male subjects per group (age range 21-35 years) were injected with 4 ng/kg, i.v., reference standard Escherichia coli (0113:h10:k) endotoxin or physiological saline. 3. Plasma inflammatory markers (tumour necrosis factor (TNF)-alpha, interleukin (IL)-6 and TNF-receptor I (RI)) and cell-adhesion molecule markers (soluble L-selectin, soluble P-selectin, soluble vascular cell adhesion molecule (VCAM)-1) were determined using sensitive and specific ELISA. 4. Tumour necrosis factor-alpha increased from a basal level of 2.8 pg/mL to approximately 800 pg/mL at 90 min after endotoxin. Similarly, IL 6 peaked 2-3 h after endotoxin injection, with a rapid decline by 6-8 h, and levels returned to basal values by 24 h. 5. In contrast, TNF-RI peaked at 2 h (increasing from basal levels of 900-3300 pg/mL) with a much slower decline and without return to basal levels at 24 h (1400 pg/mL). 6. Endotoxin resulted in a rapid rise in soluble L-selectin within 1 h, which increased from a basal of 150 425 ng/mL. This rapid rise in soluble L-selectin was sustained for up to 2.5 h and then rapidly declined to basal levels by 3.5 h. 7. In contrast, plasma soluble P-selectin levels showed a delayed and progressive increase up to 8 h (increasing from a basal level of 50-95 ng/mL), with a partial decline at 24 h (80 ng/mL). 8. Similarly, soluble VCAM-1 levels showed a progressive rise up to 24 h (increasing from basal values of 600-1000 ng/mL). 9. This acute human model of endotoxin exposure demonstrated an upregulation of inflammatory stimuli leading to a short-term hyperactivation of leucocytes and a more sustained activation of platelets and endothelium. 10. This model provides a non-invasive method for studying the complex effects of endotoxin-like pathogens on different cellular events using soluble plasma surrogate markers. PMID- 11380510 TI - Effects of digoxin, furosemide, enalaprilat and metoprolol on endothelial function in young normotensive subjects. AB - 1. Endothelial dysfunction is seen in patients with essential hypertension or congestive heart failure (CHF). The present study aimed to evaluate the direct effect on endothelium- dependent vasodilation (EDV) of different pharmacological drugs commonly used in the treatment of these conditions. 2. Forearm blood flow (FBF) was measured in 37 young healthy normotensive subjects with venous occlusion plethysmography during local intra-arterial infusions of methacholine (MCh; 2-4 microg/min), evaluating EDV, and sodium nitroprusside (SNP; 5-10 microg/min), evaluating endothelium-independent vasodilation (EIDV). The measurements of EDV and EIDV were undertaken under baseline conditions and were repeated after 1 h intra-arterial infusion of digoxin (0.1 mg/h), furosemide (5.0 mg/h), enalaprilat (2,4 mg/h), metoprolol (1.2 mg/h) or saline (controls). 3. Enalaprilat and digoxin improved the FBF response to MCh at 4 microg/min (from 22.7+/-2.3 to 25.5+/-2.1 mL/min per 100 mL tissue (P < 0.01) and from 18.2+/-2.4 to 22.2+/-2.0 mL/min per 100 mL tissue (P < 0.05), respectively). No significant changes where induced by furosemide or metoprolol in response to MCh at 4 microg/min (from 19.4+/-2.0 to 22.9+/-2.8 and from 15.3+/-2.4 to 14.7+/-1.1 mL/min per 100 mL tissue, respectively). No significant changes in basal FBF or EIDV were induced by the different drugs. When the endothelial function index was calculated as the MCh: SNP FBF ratio, a significant improvement was seen only with enalaprilat (1.1+/-0.1 to 1.2+/-0.1; P < 0.01) and furosemide (1.0+/-0.1 to 1.3+/-0.4; P < 0.05). 4. In conlusion, the results of the present study show that enalaprilat and furosemide improve endothelial vasodilatory function, while no major effect was induced by digoxin or metoprolol. Thus, different direct effects on the endothelium in young normotensive subjects were induced by drugs commonly used in the treatment of hypertension or CHF. PMID- 11380511 TI - Effect of a series of novel sulphonylthioureas on glucose tolerance in the obese fa/fa Zucker rat. AB - 1. The recent development of a series of novel KATP channel modulators, namely sulphonylthioureas and sulphonylureas, is thought to make improvements in potency and tissue selectivity compared with current sulphonylureas, such as glibenclamide, which shares a similar structure to the novel compounds. 2. These novel compounds were first examined for their effect on hyperglycaemia and glucose tolerance during an oral glucose tolerance test following 5 days administration in the lean fa/- and obese fa/fa Zucker rat (a model of insulin resistance). Comparisons with present antidiabetic agents, metformin and glibenclamide were performed. 3. Several compounds showed improvements in glucose tolerance compared with control and the primary structural prerequisites for the maintenance of this activity were investigated. Of most interest was compound 3 15 ((N-[(4-methylphenylsulphonyl]-N'-(2-ethoxypyrid-4-yl)thiourea; 0.1 mg/kg per day), which significantly improved glucose tolerance following 5 days administration in the fa/fa Zucker rat. This paralleled the improvement seen in metformin (300 mg/kg per day)-treated fa/fa rats, but compound 3-15 was up to 3000-fold more potent than metformin. 4. Obese fa/fa Zucker rats were then treated with compound 3-15 for 28 days to determine whether glycaemic control could be maintained over the longer term. 5. Compound 3-15 showed a significant improvement in glucose clearance and reduction in insulin concentration following 28 days treatment during an intravenous glucose tolerance test compared with untreated rats, without any change in the rate of weight gain. 6. The novel sulphonylthiourea 3-15 appears to improve glucose clearance during acute and chronic treatment in the fa/fa Zucker rat with no effect on the rate of weight gain. It is thought that compound 3-15 may be eliciting its actions by improving insulin sensitivity, but its effects on insulin secretion are still to be elucidated. PMID- 11380512 TI - Role of calcitonin gene-related peptide in ischaemic preconditioning in diabetic rat hearts. AB - 1. It has been suggested that calcitonin gene-related peptide (CGRP) is involved in the protection provided by ischaemic preconditioning in rat hearts and that ischaemic preconditioning is absent in diabetic rat hearts. 2. In the present study, we tested the relationship between sensory nerve function and ischaemic preconditioning in diabetic rats. 3. In 4- and 8-week diabetic rats and age matched non- diabetic controls, 30 min global ischaemia and 40 min reperfusion caused a significant decrease in cardiac function and a marked increase in creatine kinase (CK) release. Ischaemic preconditioning, by three cycles of 5 min ischaemia and 5 min reperfusion, improved the recovery of cardiac function and decreased CK release during reperfusion in 4-week diabetic rat hearts. However, the cardioprotection afforded by ischaemic preconditioning was lost in 8-week diabetic rat hearts. Pretreatment with CGRP for 5 min also significantly improved the recovery of cardiac function and decreased CK release in rats subjected to 4 or 8 weeks of diabetes. 4. The content of CGRP in the coronary effluent during ischaemic preconditioning was significantly increased in 4-week diabetic rat hearts (P < 0.05). However, only a slight increase in the release of CGRP was shown in 8-week diabetic rat hearts (P > 0.05). 5. In summary, the present results suggest that the protection afforded by ischaemic preconditioning is attenuated in diabetic rats and that the change may be related to the reduction in CGRP release in diabetic rat hearts. PMID- 11380513 TI - Co-release of endogenous ATP and noradrenaline from guinea-pig mesenteric veins exceeds co-release from mesenteric arteries. AB - 1. The present study was designed to compare the overflow of sympathetic neurotransmitters of guinea-pig inferior mesenteric artery and mesenteric vein evoked by electrical field stimulation (EFS) with special emphasis on the simultaneous release of ATP and noradrenaline (NA). The stimulation-evoked overflow of ADP, AMP and adenosine was also evaluated. 2. Endothelium-denuded segments of inferior mesenteric arteries or veins were superfused in a small volume (200 microL)-chamber for EFS and subsequent detection of NA (HPLC- electrochemical detection) and adenine nucleotides and adenosine (HPLC fluorescence detection) in samples of the superfusate. 3. Both arteries and veins responded to EFS (15 V, 4-16 Hz, 0.3 msec for 60 s) with overflow of ATP and NA in a tetrodotoxin (1 micromol/L)- and guanethidine (10 micromol/L)-sensitive manner. The EFS-evoked overflow of NA in veins exceeded the overflow of NA in arteries at all frequencies of stimulation, whereas the EFS-evoked overflow of ATP, ADP and AMP in veins exceeded the overflow of adenine nucleotides in arteries at 4 and 8 Hz but not at 16 Hz stimulation. The EFS-evoked overflow of adenosine was similar in arteries and veins. 4. Activation of alpha1 adrenoceptors with methoxamine (10 micromol/L) did not produce overflow of ATP. 5. Blockade of alpha1/alpha2-adrenoceptors with phentolamine (1 micromol/L) did not affect EFS-evoked overflow of ATP, ADP, AMP and adenosine. 6. It is concluded that overflow of ATP and NA from sympathetic nerves may constitute an effective mechanism in the complex balance between capacitance and resistance in splanchnic circulation. PMID- 11380514 TI - Pharmacological characterization of bradykinin B1 and B2 receptors in IMR-90 and INT-407 human cell lines using a microphysiometer. AB - 1. In the present study, we used a microphysiometer to measure bradykinin-induced acidification responses in IMR-90, a human lung fibroblast cell line, and INT 407, a human colonic epithelial cell line. Furthermore, we investigated the effect of 24 h exposure of transforming growth factor (TGF)-alpha on the bradykinin response in INT-407 cells. 2. Bradykinin (0.1-100 nmol/L) was potent in producing acidification responses in IMR-90 cells (pEC50 8.79+/-0.13; Hill slope 0.96+/-0.04) and INT-407 cells (pEC50 8.90+/-0.04; Hill slope 1.00+/-0.07). These responses were competitively antagonized by the bradykinin B2 receptor antagonist icatibant in both IMR-90 cells (apparent pKB = 8.54+/-0.15; Hill slope = 1.09+/-0.13 and 1.66+/-0.26 in the absence and presence of 10 nmol/L icatibant, respectively) and INT-407 cells (pKB = 8.12+/-0.07 (3, 10 and 30 nmol/L icatibant); Hill slope = 1.06+/-0.04). However, the bradykinin B1 receptor antagonist des-Arg9Leu8-bradykinin (3 micromol/L) had no effect on the bradykinin responses. 3. The non-peptide bradykinin B2 receptor antagonist FR173657 selectively antagonized bradykinin-induced acidification responses in INT-407 cells in a competitive manner (pKB = 8.76+/-0.10; Hill slope = 0.92+/-0.05) at lower concentrations (1 and 3 nmol/L) but in an insurmountable manner at higher concentrations (10 nmol/L; Hill slope = 1.04+/-0.09). This compound, at concentrations of 10 and 100 nmol/L (Hill slope = 1.38+/-0.15), also proved to be an insurmountable antagonist in IMR-90 cells. 4. The bradykinin B1 receptor selective agonist Lys0des-Arg10-bradykinin (0.1 nmol/L to 0.1 micromol/L) failed to produce acidification responses in IMR-90 cells, even after 24 h pre incubation with bacterial lipopolysaccharide (0.1 microg/mL). 5. A 24 h pre incubation of INT-407 cells with TGF-alpha (1, 10 and 100 ng/mL) caused a significant concentration-dependent decrease in maximal bradykinin response without affecting the pEC50. 6. In addition to this study being the first to use a microphysiometer to characterize bradykinin B2 receptors in cultured IMR-90 human lung fibroblast cells and INT-407 human colonic epithelial cells, we also showed that pre-incubation of INT-407 cells with TGF-alpha caused a significant decrease in maximal acidification response mediated by bradykinin B2 receptors. PMID- 11380515 TI - Effect of anti-oxidant treatment and cholesterol lowering on resting arterial tone, metabolic vasodilation and endothelial function in the human forearm: a randomized, placebo-controlled study. AB - 1. The aim of the present study was to determine whether anti-oxidant therapy with vitamin E and/or cholesterol-lowering therapy with simvastatin would augment resting forearm blood flow (FBF) and metabolic vasodilation in response to exercise and improve endothelial function in young patients with hypercholesterolaemia. 2. Endothelium-dependent and -independent, nitric oxide (NO)-mediated vasodilation have been shown to be impaired in young, otherwise healthy subjects with hypercholesterolaemia. Recent experimental and clinical studies suggest that vascular function may be improved with anti-oxidant or cholesterol- lowering therapy, although these treatments may be synergistic. 3. We compared FBF at rest, in response to isotonic exercise, the endothelium dependent vasodilator acetylcholine (ACh), the endothelium-independent vasodilator sodium nitroprusside (SNP) and the NO synthase inhibitor NG monomethyl-L-arginine (L-NMMA) in 26 young, otherwise healthy volunteers (mean (+/-SD) age 29+/-7 years; 14 female, 12 male) with hypercholesterolaemia, before and after 6 months treatment with vitamin E, simvastatin and/or placebo. Treatment was randomized, double-blinded in a 2 x 2 factorial design. Forearm blood flow was measured using venous occlusion plethysmography. 4. Vitamin E therapy increased plasma alpha-tocopherol from 39.5+/-9.6 to 75.7+/-33.8 micromol/L (P < 0.001). Simvastatin reduced total cholesterol from 6.9+/-1.7 to 4.9+/-0.8 mmol/L and low- density lipoprotein (LDL) from 4.8+/-1.7 to 3.0+/-0.7 mmol/L (both P < 0.001), although total and LDL-cholesterol also decreased slightly in the placebo group. Vitamin E increased resting FBF from 2.1+/-0.3 to 2.4+/-0.3 mL/100 mL per min (P = 0.04) and decreased resting forearm vascular resistance from 42.1+/-4.2 to 36.1+/-3.4 units (P = 0.01), but the reduction in resting FBF with L-NMMA was not affected. Vasodilation in response to isotonic exercise, ACh and SNP was similar before and after treatment in the placebo, vitamin E, simvastatin and in the combined vitamin E-simvastatin groups. NG Monomethyl-L-arginine infusion reduced resting FBF and functional hyperaemia in response to exercise and these responses were not altered by treatment. 5. These data suggest that while vitamin E therapy augments resting FBF and reduces forearm vascular resistance in young hypercholesterolaemic subjects, these effects may not be via NO-dependent pathways. Metabolic vasodilation and responses to the NO-mediated vasodilators ACh and SNP were not favourably affected by anti-oxidant or cholesterol-lowering therapy, either alone or in combination. PMID- 11380516 TI - Multiple inert gas elimination technique for determining ventilation/perfusion distributions in rat during normoxia, hypoxia and hyperoxia. AB - 1. The use of the multiple inert gas elimination technique (MIGET) in quantifying ventilation/perfusion distributions (V*A/Q*) in small animals, such as the rat, may cause results to be biased due to haemodilution produced by the large volume of liquid infused intravenously. 2. We tested two methods of administering inert gases in rats using the MIGET: (i) standard continuous intravenous administration of inert gases (method A); and (ii) a new method based on the physicochemical properties of each inert gas (method B). This method included acute simultaneous inert gas administration using three pathways: inhalation, intravenous infusion and rectal infusion. Both MIGET methods were applied to obtain data while breathing three different inspiratory fractions of oxygen (FIO2): normoxia, hypoxia and hyperoxia. 3. Inert gas levels obtained from blood or expired air samples were sufficient for chromatographic measurement, at least during a 2 h period. The V*A/Q* distributions reported using both methods were acceptable for all the physiological conditions studied; therefore, the alternative method used here may be useful in further MIGET studies in rats because haemodilution resulting from continuous intravenous infusion of less-soluble gases can be avoided. 4. Normoxic rats showed lower mean values of the V*A/Q* ratio of ventilation distribution and higher mean values of the V*A/Q* ratio of perfusion distribution with the usual method of inert gas administration (method A). These non-significant differences were observed under almost all physiological conditions studied and they could be caused by haemodilution. Nevertheless, the effect of interindividual differences cannot be discarded. An additional effect of the low haematocrit on cardiovascular changes due to low FIO2, such as pulmonary vasoconstriction or increased cardiac output, may explain the lower dispersion of perfusion distributions found in group A during hypoxia. PMID- 11380517 TI - Cyclo-oxygenase 2 tissue distribution and developmental pattern of expression in the chicken. AB - 1. Cyclo-oxygenase 2 (COX-2) is implicated in multiple physiopathological processes. We have studied its physiological expression during chicken embryogenesis. 2. An original procedure was set to prepare a COX-2 probe from red blood cells. In situ hybridization, reverse transcription-polymerase chain reaction and northern blots, were performed on chick embryos from embryonic day (E) 3 to postnatal day 15. 3. In the mesonephros, the signal detected in mesonephric tubules presented a slow increase from E5 to E9, a plateau up to E12 and then a decrease, while the signal increased in the metanephros and then decreased after hatching. Transient expression of COX-2 mRNA in endothelial cells of the infundibulum of the aorta was also detected between E12 and E17. 4. This enzyme may have important roles in kidney morphogenesis during early embryonic stages and in tubular functions during development and in adult life. In the cardiovascular system, its vasodilatory effect could modulate the vasoconstrictor effect of the systolic pressure between E12 and E17 and contribute to a normal morphogenesis of the arterial tree and heart. PMID- 11380518 TI - Significance of exaggerated natriuresis after angiotensin AT1 receptor blockade or angiotensin- converting enzyme inhibition in obese Zucker rats. AB - 1. Obese Zucker rats (OZR) were shown to be salt-sensitive in that they develop hypertension when placed on a high-salt diet. Because angiotensin (Ang) II is a major antinatriuretic factor, the present studies were undertaken to determine whether the characteristic of salt-sensitivity of OZR is associated with an enhanced antinatriuretic function of endogenous AngII. 2. The extent of AngII mediated antinatriuresis was investigated in OZR and lean Zucker rats (LZR) using candesartan (100 microg/kg, i.v.), a selective angiotensin AT1 receptor antagonist, and ramipril (1 mg/kg, i.v.), an angiotensin-converting enzyme (ACE) inhibitor. The total number of AngII binding sites and their affinity were also assessed in renal cortical tubular membrane preparations of OZR and LZR using a specific radioligand-binding assay. Plasma renin activity was determined using a standard radioimmunoassay. 3. Both candesartan and ramipril produced substantially greater increases in urinary sodium excretion and urine flow in OZR and these effects were significantly greater than those observed in LZR. These observations suggest that basal antinatriuretic function of endogenous AngII is exaggerated in OZR. 4. The functional overexpression of AngII was not due to any alterations in the affinity or the total number of AngII binding sites in renal cortical tubular membranes. Higher plasma renin values in the OZR could have contributed to the phenomenon. 5. In conclusion, marked diuresis and natriuresis after AT1 receptor blockade and/or ACE inhibition suggest that the extent of endogenous AngII-mediated sodium transport under basal conditions is greatly augmented in OZR. It is proposed this phenomenon may be a contributing factor for the salt- sensitivity in the OZR. PMID- 11380519 TI - K(Ca) channel-opening activity of Ginkgo Biloba extracts and ginsenosides in cultured endothelial cells. AB - 1. Extracts of Ginkgo biloba (EGb) and ginsenosides (GS) have been reported to induce vasorelaxation. In the present study, the role of K+ channels in the action of EGb and GS to activate nitric oxide synthase (NOS) activity was investigated in cultured endothelial cells. 2. Nitric oxide synthase activity of cultured endothelial cells detected by the reduced nicotinamide adenine dinucleotide phosphate (NADPH) histochemistry method was significantly increased after treatment with 20 microg/mL EGb or 40 microg/mL GS plus 10 mmol/L L arginine. The effect was completely abolished by the addition of 0.5 micromol/L Nomega-nitro-L-arginine, an inhibitor of NOS, to the incubation medium and partially inhibited by 10 micromol/L tetraethylammonium (TEA), an inhibitor of Ca2+-activated K+ (KCa) channels. 3. Application of EGb to the intracellular surface of excised inside-out patches activated K+ channels in a concentration dependent manner in the concentration range 1-100 microg/mL. Channel activity was also activated by application of GS at concentrations ranging from 1 to 300 microg/mL. The modulation of channel activity was inhibited by 0.5 mmol/L TEA but not by 0.5 mmol/L glibenclamide, an inhibitor of ATP-sensitive K+ channels. 4. Thus, in cultured endothelial cells, the increase in NOS activity induced by EGb or GS depends on the activity of KCa channels. These compounds may regulate nitric oxide release by changing the cell membrane potential in vascular endothelial cells. PMID- 11380520 TI - Comparison of digoxin versus low-dose amiodarone for ventricular rate control in patients with chronic atrial fibrillation. AB - 1. Rapid ventricular rate (VR) and rhythm irregularity during atrial fibrillation (AF) impair cardiac performance. Although digoxin has been widely used in patients with AF, its efficacy for the control of VR and rhythm irregularity is unsatisfactory. Whether low-dose amiodarone is more effective remains unclear. 2. We randomized 16 patients (13 male, three female; mean (+/-SD) age 63 +/- 9 years) with chronic AF to receive either digoxin or amiodarone for 24 weeks. At baseline and at 12 and 24 weeks follow up, Holter monitor recording and cardiopulmonary exercise test were performed to assess VR and rhythm irregularity control and exercise capacity. 3. Seven and nine patients received digoxin and amiodarone, respectively. After 12 and 24 weeks treatment, both digoxin and amiodarone significantly decreased the mean ambulatory VR and the VR during peak exercise compared with baseline (all P < 0.05). At 24 weeks, there were no significant differences between digoxin and amiodarone in the percentage reduction in VR during ambulatory (27 +/- 13 vs. 25 +/- 12%, respectively; P = 0.8) and peak exercise (13 +/- 12 vs. 12 +/- 10%%, respectively; P = 0.6). 4. The rhythm irregularity, as measured by SD of RR intervals and the root mean square of the SD of RR intervals, and the exercise capacity, as measured by exercise workload, maximal oxygen consumption (VO2), minute ventilation, ventilatory equivalent and oxygen pulse, were not significantly changed after treatment with digoxin or amiodarone (all P > 0.05). 5. Quality of life, determined by SF-36 questionnaire, and AF symptomatology, as measured by the AF Symptom Checklist, were also not significantly changed after treatment with digoxin or amiodarone (all P > 0.05). 6. In conclusion, digoxin and low-dose amiodarone had similar efficacy in the control of VR during ambulatory activity and exercise. However, both were less efficacious during exercise and did not significantly affect rhythm irregularity, exercise capacity, quality of life and AF symptomatology in patients with chronic AF. PMID- 11380521 TI - An electrophysiological study of the effects of propofol on native neuronal ligand-gated ion channels. AB - 1. Pharmacological evidence suggests that some of the clinical actions of propofol may be mediated, at least in part, by positive modulation of the GABA(A) receptor chloride channel. The effect of propofol at other native neuronal ligand gated ion channels is unclear. 2. To gain some insight into the effects of propofol at a range of native neuronal receptors, the present study has used an extracellular recording technique and determined its effects at GABA(A), 5-HT3, P2X and nicotinic acetylcholine (nACh) receptors of the rat isolated vagus nerve and the GABA(A) and strychnine-sensitive glycine receptor of the rat isolated optic nerve. In addition, we have used patch-clamp recording techniques to further investigate the effects of propofol at the GABA(A) and strychnine sensitive glycine receptors in rat cultured hippocampal neurons. 3. Propofol (0.3 100 micromol/L) concentration-dependently potentiated submaximal GABA-evoked responses in the vagus nerve and shifted the GABA concentration-response curve to the left. In contrast, propofol at concentrations ranging from 1 to 10 micromol/L had little or no effect on 5-HT3, P2X or nACh receptor-mediated responses in the vagus nerve but, at 100 micromol/L, propofol inhibited these responses to approximately 50% of control. In the optic nerve, EC20 GABA-evoked responses were also potentiated by propofol (10 micromol/L), while EC20 glycine-evoked responses were minimally enhanced. 4. Further investigations using cultured hippocampal neurons showed that submaximal (10 micromol/L) GABA-evoked currents were potentiated by propofol (1-10 micromol/L), in a non-voltage-dependent manner, whereas submaximal (100 micromol/L) glycine-evoked currents were unaffected. 5. These data suggest that propofol, at therapeutic concentrations, exerts its principle pharmacological actions at GABA(A) receptors with relatively little effect at other neuronal ligand-gated ion channels. PMID- 11380522 TI - Nitric oxide reduces pressor responsiveness during ovine hypoadrenocorticism. AB - 1. Hypoadrenocorticism frequently results in critical hypotension, hypovolaemia, hyponatraemia and hyperkalaemia. Perhaps even more important, hypoadrenocorticoid humans experience decreased vasoconstriction in response to exogenous administration of vasoconstrictors, such as noradrenaline. 2. We studied chronically adrenalectomized adult sheep to test the hypothesis that the reduction in pressor responsiveness is the result of increased production of nitric oxide (NO) during hypoadrenocorticism. 3. Withdrawal of steroid replacement resulted in reduced blood pressure, reduced pressor responsiveness, as well as hyperkalaemia and hyponatraemia. 4. Inhibition of NO production by NG nitro-L-arginine methyl ester in the hypoadrenocorticoid ewes restored mean arterial pressure and pressor responsiveness response to normal values. 5. The results of these experiments support the hypothesis that reduced pressor responsiveness in the hypoadrenocorticoid state is mediated by the overproduction of NO. PMID- 11380523 TI - Non-invasive single cell pH measurements in the isolated perfused pancreas. AB - 1. A new method was developed for non-invasive investigations of intracellular pH (pHi) regulation in different cell types of the isolated perfused pancreas using a confocal laser scanning technique. 2. After removal of the rat pancreas the coeliac artery was cannulated and the splenic segment of the pancreas was perfused with dextran (5%)-Ringer solution at a constant flow rate of 2 mL/min. In a temperature-controlled (37 degrees C) chamber, pH regulation was studied using the pH-sensitive fluorescent dye 2',7'-bis-(2-carboxyethyl)-5-(-6) carboxyfluorescein (BCECF) with a confocal microscope (MRC-600; Bio-Rad, Hercules, CA, USA). 3. Image analysis permitted the identification and comparison of different cell types with a pHi of 7.26+/-0.1 in acinar cells and of 7.02+/ 0.1 in endothelial cells. Increasing PCO2 from 5 to 20% resulted in a rapid decrease in pHi. Omission of sodium from the perfusate resulted in a smooth decline in pHi. Both decreases were found to be fully reversible. Increasing PCO2 under sodium-free conditions also resulted in a drop of pHi that was, however, not fully reversible, suggesting involvement of the Na+/H+ exchanger in the regulation of pHi in the intact organ. 4. The above method completely preserves tissue integrity and, therefore, allows the study of pH regulation in different cell types of the pancreas simultaneously and without interference with their functional arrangement. The technique should be of specific value to investigate experimental disease states of the pancreas. PMID- 11380524 TI - A proteasome inhibitor prevents vascular hypertrophy in deoxycorticosterone acetate-salt hypertensive rats. AB - 1. In the present study, we investigated the potential of the proteasome inhibitor N-benzyloxycarbonyl-Ile-Glu(O-t-Bu)-Ala-leucinal (PSI) to prevent vascular hypertrophy induced by deoxycorticosterone acetate (DOCA) and salt in rats. 2. Vehicle (35% ethanol, 35% polyethylene glycol and 30% saline solution) treated DOCA-salt rats developed marked hypertension at 4 weeks. Morphological studies on the rats given vehicle showed aortic hypertrophy, with a significant increase in wall thickness, wall area and wall-to-lumen ratio. A significant decrease in vascular wall hypertrophy was observed in PSI (3 mg/kg)-treated DOCA salt rats. In addition, a marked increase in aortic endothelin (ET)-1 content was evident in vehicle-treated DOCA-salt rats compared with findings in sham-operated rats. A significant attenuation of this increase occurred in PSI-treated DOCA salt rats. 3. These results indicate that PSI can prevent the vascular hypertrophy in DOCA-salt hypertensive rats and the effect is accompanied by suppression of ET-1 production in the aorta. We suggest that a proteasome dependent proteolytic system has an important role in the development of vascular hypertrophy in cases of DOCA-salt-induced hypertension, possibly through the enhancement of ET-1 production in vascular tissues. PMID- 11380525 TI - Temporal age-related changes in spectral, fractal and complexity characteristics of heart rate variability. AB - Cross-sectional studies have suggested that heart rate (HR) variability, analysed using traditional time and frequency domain methods, is related to ageing, but no longitudinal studies have estimated the age dependence of HR fluctuation. This study evaluated temporal age-related changes in 12-h measures of HR variability among 109 patients with coronary artery disease (CAD), who underwent repeat Holter recordings at 32-month intervals. Time and frequency domain measures, along with fractal and complexity measures of HR variability, were determined at the baseline and after 32 months. Changes in HR dynamics were compared with various laboratory variables, exercise data and angiographic progression of CAD. Traditional time and frequency domain measures of HR variability did not change significantly during the follow-up, but the power-law scaling slope decreased from -1.29 +/- 0.20 to -1.36 +/- 0.23 (P<0.01) and the short-term fractal exponent (alpha1) of HR dynamics from 1.29 +/- 0.14-1.22 +/- 0.18 (P<0.001). The approximate entropy value also decreased from 1.00 +/- 0.19 to 0.95 +/- 0.18 (P<0.05). The changes in HR behaviour were not related to demographic data, laboratory values or angiographic progression of CAD. Only a weak correlation was observed between the change in the power-law slope and the baseline glucose value (P<0.05). This longitudinal study shows that the fractal characteristics of HR dynamics and the complexity properties of R-R intervals undergo rapid changes along with ageing, and that fractal and complexity analysis techniques are more sensitive than traditional analysis methods in documenting temporal age-related changes in HR behaviour. PMID- 11380526 TI - Air pollutants and tear film stability--a method for experimental evaluation. AB - Instability of the pre-ocular tear film causes symptoms of dry eyes. Tear film instability may be caused by exposure to air pollutants, but identification of causative agents is often difficult. We describe an in vitro technique for assessment of putative agents on the surface activity of Meibomian lipids. The lipids were obtained from bovine eyelids and dissolved in chloroform. Surface activity of the solution was assessed on a Wilhelmy balance. After addition of benzalkonium chloride, minimum surface tension of the lipids increased and maximum surface tension fell. After addition of quartz particles, both maximum and minimum surface tension increased. The method can be used to screen substances suspected to cause dry eyes, providing a basis for selection of substances for in vivo studies. PMID- 11380527 TI - Sympathetic skin vasoconstriction--further evaluation using laser Doppler techniques. AB - The aim of this study was to quantify the reflex sympathetic vasoconstriction in skin at different depths. Twenty healthy subjects were studied. Finger skin blood flow was measured using laser Doppler perfusion imaging (LDPI) and laser Doppler perfusion monitoring (LDPM). In LDPM, a probe with fibres separated 0.25 mm (deep) and 0.14 mm (superficial) from the illuminating fibre was used. Local heating (40 degrees C) was achieved with a Peltier element, and reflex vasoconstriction induced by immersion of the contra-lateral hand and forearm for 3 min in water at 15 degrees C. The change in skin blood flow was measured and a vasoconstriction index (VAC: cooling/before cooling) calculated. VAC indices of LDPI, LDPM-0.25 and LDPM-0.14 were 0.60, 0.59 and 0.60, respectively. The two components of the LDPM perfusion value, blood cell velocity and concentration, were studied separately. Their contributions in LDPM-0.25 were roughly the same, whereas the velocity component dominated in LDPM-0.14, although their relative responses in the two channels were similar. We conclude that sympathetic skin vasoconstriction does not significantly differ in two compartments, as probed with fibres separated by 0.25 and 0.14 mm. Blood cell velocity is influenced in a proportional way, as is concentration. PMID- 11380528 TI - Effects of exogenous melatonin on pituitary hormones in humans. AB - The effects of melatonin on physiological function remain unclear, although the therapeutic potential of melatonin is being increasingly recognized. The aim of the present study is to investigate the effects of exogenous melatonin on the spontaneous release of pituitary hormone in humans. A double blind placebo controlled protocol was designed to examine 12 adult healthy volunteers and 12 sleep disorder patients who have been treating with low doses of melatonin for 1 year. Either exogenous melatonin or placebo of 1 mg was given at 09:00 hours, followed by the collection of blood samples every 20 min for 4 h. Each blood sample was examined for levels of serum melatonin, PRL, LH, FSH, GH and TSH. LH levels were higher in sleep disorder patients compared with the healthy volunteers. In other pituitary hormones, there were no significant difference between healthy adults and sleep disorder patients. In all subjects, PRL levels were stimulated by acute administration of 1 mg of exogenous melatonin, while the levels of other pituitary hormones were not affected. These results suggested that exogenous melatonin can affect the spontaneous release of LH and PRL in humans. In addition, we demonstrated that 1-year oral melatonin treatment did not affect the responses to the acute administration of melatonin. PMID- 11380529 TI - Postprandial impairment of resistance vessel function in insulin treated patients with diabetes mellitus type-2. AB - Reduced postischaemic reactive hyperaemia, is considered a marker of impaired resistance vessel function. Acute postprandial hyperlipidaemia has been shown to induce vascular dysfunction. In the present study, the impact of postprandial hyperglycaemia on resistance vessel reactivity was investigated in insulin treated type-2 diabetic patients. The study was performed in 16 insulin treated type-2 diabetics (eight male/eight female, age 47 +/- 3 years, HbA1c 7.2 +/- 0.2) and 16 controls. Reactive hyperaemia was measured in the forearm by venous occlusion plethysmography after 5 min of ischaemia in the fasting state and 90 min after a test meal. In diabetics, blood glucose increased from 8.7 +/- 1.1 to 15.3 +/- 1.0 mmol l-1 (P<0.001) postprandially. This resulted in (i) a significant increase of resting blood flow (3.4 +/- 0.3 to 4.8 +/- 0.4 ml min-1 100 ml-1, P<0.01) and (ii) in a reduced peak reactive hyperaemia (52.3 +/- 7.4 to 36.8 +/- 4.3 ml min-1 100 ml-1, P<0.005). In controls, a similar effect of the meal on resting flow was observed but reactive hyperaemia was unaltered. In the absence of a test meal, basal flow as well as peak reactive hyperaemia remained unchanged in diabetic as well as in non-diabetic subjects. Our data provide evidence that in the postprandial state resistance vessel reactivity becomes reduced in insulin treated type-2 diabetic patients. PMID- 11380530 TI - The changes in leg blood flow during and after mild or severe acute hypoxaemia in healthy humans. AB - The present study examines the leg blood flow changes in resting healthy humans during and after a 10-min period of mild (PaO2=5.60 kPa) or severe hypoxaemia (PaO2=4.53 kPa) induced by breathing hypoxic gas mixtures. A Colour Duplex Scan system allowed to measure the cross-sectional area (CSA) and mean blood flow (Q) in a femoral artery (FA) and a femoral vein (FV) and also in an artery supplying leg muscles (medial gastrocnemius artery, MGA). During the mild as well as the severe hypoxaemia and their recovery periods, no significant variations of Q and CSA occurred in FA and FV. During the mild hypoxaemia and the first 10 min of the recovery period, Q and CSA of MGA increased (maximal changes: +84 and +20%, respectively). By contrast, a marked Q decrease and a reduced CSA were measured in MGA during the severe hypoxaemia (-67 and -60%, respectively). This reduced muscle blood flow was followed by a vasodilatation (CSA increase = +30%), which began 10 min after the hypoxaemia ended and persisted for a further 10-min period. This study shows that the time course of muscle blood flow changes in response to acute hypoxaemia depends on the PaO2 level. Reverse effects were measured during the mild or the severe hypoxaemia, whereas a post-hypoxaemic vasodilatation occurred in all circumstances. PMID- 11380531 TI - Long-term reproducibility of ambulatory blood pressure in unselected elderly subjects. AB - The aim of this study was to evaluate the long-term reproducibility and validity of 24-h ambulatory blood pressure measurements (ABPM) in an unselected elderly population. In a rural Finnish community 503 randomly chosen invited persons over 65 years of age participated and went through 24-h ABPM. As part of the validation of the methodology, the reproducibility study was conducted in 26 persons (age 65-76 years). Two identical sets of measurement were performed at 4 12 (median 8) month intervals. The agreement between measurements was assessed by correlation coefficients and standard deviation (SD) of the differences. There were no significant differences in 24-h, daytime and night-time average diastolic blood pressure (DBP) and daytime average systolic blood pressure (SBP) between the two measurements. During the second measurement, 24-h SBP and night-time average SBP were slightly higher than those obtained by the first monitoring. Average 24-h SBP and DBP were 18 and 7 mmHg lower, respectively, than office blood pressure averages. The correlation coefficients were significantly higher for 24-h ambulatory blood pressure than for office blood pressure. The SD of the mean difference between visits was significantly lower for 24-h ambulatory blood pressure than for office blood pressure measurements. These findings show that the long-term reproducibility of ambulatory blood pressure is good in an elderly unselected population and better than the office blood pressure reproducibility. PMID- 11380532 TI - Influence of respiratory motor neurone activity on human autonomic and haemodynamic rhythms. AB - Although humans hold great advantages over other species as subjects for biomedical research, they also bring major disadvantages. One is that among the many rhythmic physiological signals that can be recorded, there is no sure way to know which individual change precedes another, or which change represents cause and which represents effect. In an attempt to deal with the inherent complexity of research conducted in intact human subjects, we developed and used a structural equation model to analyse responses of healthy young men to pharmacological changes of arterial pressure and graded inspiratory resistance, before and after vagomimetic atropine. Our model yielded a good fit of the experimental data, with a system weighted R2 of 0.77, and suggested that our treatments exerted both direct and indirect influences on the variables we measured. Thus, infusions of nitroprusside and phenylephrine exerted all of their direct effects by lowering and raising arterial pressure; the changes of R-R intervals, respiratory sinus arrhythmia and arterial pressure fluctuations that these drugs provoked, were indirect consequences of arterial pressure changes. The only direct effect of increased inspiratory resistance was augmentation of arterial pressure fluctuations. These results may provide a new way to disentangle and understand responses of intact human subjects to experimental forcings. The principal new insight we derived from our modelling is that respiratory gating of vagal-cardiac motor neurone firing is nearly maximal at usual levels of arterial pressure and inspiratory motor neurone activity. PMID- 11380533 TI - Chronotropic incompetence response to exercise in congestive heart failure, relationship with the cardiac autonomic status. AB - In chronic congestive heart failure (CHF), attenuated heart rate response to exercise, a manifestation of chronotropic incompetence (CI), contributes to limiting exercise capacity. The present study was thus conducted to evaluate the respective role of chronic attenuation of cardiac vagal tone associated with depressed baroreflex sensitivity or affected cardiac sympathetic responsiveness in CHF patients with CI. Spontaneous cardiac baroreflex sensitivity (BRS) assessed by sequence method and spectral- and time-domain analysis of heart rate variability (HRV) were analysed in 21 chronic CHF patients. All patients performed a symptom-limited exercise test with measurement of gas exchange. Chronic incompetence which was defined as failure to achieve > or =80% of the heart rate reserve (%HRR) given by (HRpeak - HRrest)/(predictive maximal heart rate - HRrest) was observed in 14 (66%) patients. There was no significant difference in age, heart rate, peak oxygen uptake or left ventricular ejection fraction between the patients with and without CI. Although there was no significant difference in BRS, low frequency power of HRV in normalized units (LFnu) and SDNN were significantly lower in CI patients. Percentage of HRR correlated significantly with LFnu on 15 min (r=0.64, P<0.005) and, with LFnu on 24 h (r=0.52, P<0.01), SDNN (r=0.48, P=0.03) and SDANN (r=0.48, P=0.03), but not BRS (r=0.04, P=NS). Autonomic nervous system derangement is a complex process in CHF. The role of basal depressed cardiac sympathetic tone seems to contribute more closely than depressed baroreflex sensitivity to the impaired heart rate response to exercise frequently observed in CHF patients. PMID- 11380534 TI - Polymorphisms in the renin-angiotensin system and endothelium-dependent vasodilation in normotensive subjects. AB - BACKGROUND: Our aim was to test the hypothesis that genes encoding components in the renin-angiotensin system influence endothelial vasodilatory function. METHODS: In 59 apparently healthy, normotensive individuals, endothelium dependent vasodilation (EDV) and endothelial-independent vasodilation (EIDV) was evaluated by infusing metacholine and sodium nitroprusside into the brachial artery. Forearm blood flow was measured by venous occlusion plethysmography. The ACE insertion (I)/deletion (D) polymorphism, the T174M and M235T angiotensinogen restriction fragments length polymorphisms, the angiotensin II receptor type 1 (AT1R) A1166C, and the aldosterone synthase gene (CYP11B2) C-344T polymorphisms were analysed. RESULTS: When analysing the ACE, the two angiotensinogen and the aldosterone synthase CYP11B2 genotypes independently, no significant association with endothelial vasodilatory function was found. However, a significant reduction in endothelium-dependent vasodilation was observed in the subjects (n=9) with the ACE D allele and the angiotensinogen T174M genotype (P<0.05). Subjects with the AT1R genotype AC showed a reduction in both EDV (P=0.05) and EIDV (P=0.04) when compared with those with the AA genotype. CONCLUSIONS: The subjects with the ACE D allele in combination with the angiotensinogen T174M genotype are associated with a reduced EDV. This together with the observation that the AC AT1R genotype is associated with a reduction in both EDV and EIDV, supports the hypothesis that endothelial vasodilatory function is influenced by genes in the renin-angiotensinogen system. PMID- 11380535 TI - Ageing is not associated with a decline in neuromuscular innervation or reduced specific force in men aged 20 and 50 years. AB - The exact mechanisms responsible for the decline in strength with age are yet to be completely elucidated. Three proposed mechanisms responsible for the detrimental effect of increasing age on strength include changes in muscle mass, specific force and/or neuromuscular innervation. Thus, the purpose of this investigation was to determine if the age-related reduction in peak isometric strength was primarily associated with changes in muscle cross-sectional area, neuromuscular innervation and/or specific force. The cross-sectional area of the knee extensor muscles (QCSA) was estimated in 13 younger men (YM; 20.8 +/- 1.6 years) and eight middle-aged men (MM; 53.8 +/- 4.2 years) prior to performing a series of four maximal voluntary isometric contractions on an isokinetic dynamometer at an angle of 60 degrees knee flexion. Peak force was determined and surface electromyography was sampled from the rectus femoris muscle during each maximal voluntary contraction. The cross-sectional area of the knee extensor muscles, peak force and integrated electromyography (IEMG) were significantly lower in the MM (P<0.01). However, when peak force and peak IEMG values were corrected for QCSA, there were no significant differences between age groups. These results suggest that the reduction in peak isometric force observed in the MM was primarily associated with quantitative changes in muscle mass, rather than reduced neuromuscular innervation or specific force. Therefore, preserving muscle mass through resistance training may significantly reduce the age-associated differences in peak strength and assist in promoting quality of life and functional independence in older adults. PMID- 11380536 TI - Ageing, soft-palate tone and sleep-related breathing disorders. AB - We compared the tissue tone of the soft palate in 10 young (18-29 years) and 15 middle-aged (30-70 years) apnoeics with 10 young and 10 middle-aged non-snorers and healthy volunteers of the same ages. The obstructive sleep apnoea syndrome (OSAS) was diagnosed by means of the self-reported questionnaire about the medical history of the subjects and polysomnography (PSG) with overnight pulse oximetry. Computerized endopharyngeal myotonometry (CEM) was used to measure the tissue tone of the soft palate. The CEM study is performed while the subjects are awake. CEM records and analyses the biomechanical properties of tissue in response to a brief mechanical impact. Tissue stiffness is expressed as a frequency and elasticity as a logarithmic decrement of damped oscillation. RESULTS: The values of the tissue stiffness (13.5, SD 2.5 Hz) and elasticity (1.2, SD 0.4) of the soft palate in young adults with OSAS reveal no important differences in comparison with the tissue stiffness (12.7, SD 2.4 Hz) and elasticity (0.9, SD 0.2) in non-snoring young adults. The difference in the value of the tissue stiffness of the soft palate is significant among middle-aged patients with OSAS (20.3, SD 4.7 Hz) and non-snoring middle-aged persons (13.9, SD 2.3 Hz). A non-significant difference was observed in the value of the tissue elasticity among middle-aged patients with OSAS (1.7, SD 0.8) and non-snoring middle-aged persons (1.6, SD 0.3). The tissue tone of the soft palate in young and middle-aged healthy subjects differs by elasticity but not by stiffness. CONCLUSIONS: We found that some biomechanical properties of the tissues of the soft palate may be changed by OSAS and some by age. PMID- 11380537 TI - Time domain, geometrical and frequency domain analysis of cardiac vagal outflow: effects of various respiratory patterns. AB - The purpose of this study was to compare the applicability of four different measures of heart rate variability (HRV) in the assessment of cardiac vagal outflow, with special reference to the effect of breathing pattern. The anticholinergic effects of an intravenous glycopyrrolate infusion (5 microg x kg( 1) x h(-1) for 2 h) during spontaneous and controlled (15 min(-1)) breathing rate were investigated in eight volunteers, and the effects of different fixed breathing rates (6-15-24 min(-1)) and hyperventilation in 12 subjects. Cardiac vagal activity was assessed by ECG recordings in which the following measures of HRV were computed: the high-frequency (HF) spectral component, the instantaneous RR interval (RRI) variability (SD1) analysed from the Poincare plots, the percentage of differences between successive RRIs greater than 50 ms (pNN50), and the square root of the mean squared differences of successive RRIs (RMSSD). On average, glycopyrrolate reduced the HF spectral component by 99.8%, SD1 by 91.3%, pNN50 by 100% and RMSSD by 97.0%. The change of breathing pattern from controlled to spontaneous decreased significantly the HF component and pNN50, but did not affect SD1 or RMSSD. Rapid breathing rate (24 min(-1)) decreased the HF component, but had no effects on the other measures. A controlled breathing rate is needed for a reliable assessment of cardiac vagal outflow by the spectral analysis technique. The quantitative geometrical analysis of short-term RRI variability from the Poincare plots and the time domain measure RMSSD were not significantly affected by changes in the breathing rate, suggesting that these indices are more suitable for the measurement of cardiac vagal outflow during the 'free-running' ambulatory conditions. PMID- 11380538 TI - Whole-body vibration exercise leads to alterations in muscle blood volume. AB - Occupationally used high-frequency vibration is supposed to have negative effects on blood flow and muscle strength. Conversely, low-frequency vibration used as a training tool appears to increase muscle strength, but nothing is known about its effects on peripheral circulation. The aim of this investigation was to quantify alterations in muscle blood volume after whole muscle vibration--after exercising on the training device Galileo 2000 (Novotec GmbH, Pforzheim, Germany). Twenty healthy adults performed a 9-min standing test. They stood with both feet on a platform, producing oscillating mechanical vibrations of 26 Hz. Alterations in muscle blood volume of the quadriceps and gastrocnemius muscles were assessed with power Doppler sonography and arterial blood flow of the popliteal artery with a Doppler ultrasound machine. Measurements were performed before and immediately after exercising. Power Doppler indices indicative of muscular blood circulation in the calf and thigh significantly increased after exercise. The mean blood flow velocity in the popliteal artery increased from 6.5 to 13.0 cm x s(-1) and its resistive index was significantly reduced. The results indicate that low-frequency vibration does not have the negative effects on peripheral circulation known from occupational high-frequency vibration. PMID- 11380539 TI - Assessing thoracoabdominal asynchrony. AB - The traditional method of derivation of phase difference between ribcage and abdomen breathing movements from a Lissajous plot is shown to be unsatisfactory for assessing the degree of asynchrony. The signal processing technique of cross correlation is introduced as a better, statistically based approach. Even so, examination of the latent structure of a Lissajous figure leads to the concept of movement sum and difference components along its principal axes. This more general form of analysis is used for indicating relative changes in tidal volume during postoperative monitoring of Cheyne-Stokes breathing with obstructive apnoea, as well as tracking the degree of asynchrony. The theoretical and practical limitations of inductive plethysmography calibrations are such that the proposed methods of uncalibrated non-invasive respiratory monitoring are also preferable as research tools. PMID- 11380540 TI - Labelling of skin sensitizers: the new European Dangerous Preparations Directive. AB - The new Dangerous Preparations Directive (DPD, 1999/45/EC) introduces a special labelling requirement for skin sensitizers in products that are regulated under this Directive. The packaging of products containing 0.1% of a sensitizer must bear the inscription "Contains 'name of sensitizer'. May produce an allergic reaction." The aim is to protect individuals already sensitized by providing information which enables them to avoid products containing ingredients which may elicit their allergy. However, this is only of benefit where such sensitized individuals do exist in the population. Moreover, this labelling requirement does not take into account the potency of the skin sensitizer. For each sensitizer and type of skin exposure, there will be levels below which it will not elicit allergic contact dermatitis reactions in individuals who are sensitized to that chemical. We therefore propose that within the new DPD, it should be possible to override this labelling requirement with well-documented data, to ensure that information provided to the consumer on the product label is not misleading. The current implementation in the DPD of what is in principle a good idea means that further action (legislative changes; scope for derogation) is needed if the potential benefits are not to be lost. PMID- 11380541 TI - Dermatitis in a particleboard manufacturing facility. AB - Exposure to wood dust and other chemicals used in particleboard manufacture may cause contact dermatitis. To assess the prevalence and nature of skin complaints in the refining and drying department of a particleboard manufacturing facility following introduction of a new process, workers were assessed by a physician, using a standardized questionnaire concerning symptoms, past and family history, workplace exposures, and use of protective equipment. Cutaneous examination and patch testing were then performed. Questionnaire results showed that workers complained of rash, nasal and eye irritation, as well as cough and bothersome odours. Cutaneous examination identified a heterogeneity of skin problems, with dermatitis being mainly irritant rather than allergic. Quaternium-15 (Dowicil 200) was the only allergen to which more than 1 individual reacted. Aside from the odours, exposure to wood dust could account for the other reported symptoms. Allergic reactions on patch testing were few and did not explain the dermatitis; most skin reactions were irritant. Recommendations to the company included improved dust control, through ventilation and personal hygiene measures, as well as protective clothing. The investigation highlights how the introduction of a new process may focus attention on health complaints that have been present for some time before. PMID- 11380542 TI - Skin-sensitization structure-activity relationships for aldehydes. AB - A selection of 17 aldehydes (13 sensitizing and 4 non-sensitizing), all of which possessed a benzene ring, were evaluated using structure-activity relationships (SARs). The sensitizing compounds were classified as strong, moderate or weak skin sensitizers on the basis of in vivo data. The aldehydes were grouped into 4 distinct subcategories of functionally related aldehydes that were termed aryl substituted aliphatic, aryl, aryl with special features (that can undergo metabolism) and alpha,beta-unsaturated aldehydes. It was observed that a structure-activity relationship could be derived for a subset of aldehydes that could react via the same chemical mechanism. This further supports the view that applying knowledge on reaction mechanisms to develop SAR models can provide a more accurate means of investigating and predicting the sensitization potential of structurally and functionally related chemicals. PMID- 11380543 TI - Beneficial effects of a skin tolerance-tested moisturizing cream on the barrier function in experimentally-elicited irritant and allergic contact dermatitis. AB - In experimentally-induced irritant (ICD) and allergic (ACD) contact dermatitis, an oil-in-water (o/w) cream was applied to investigate its effects on a disturbed barrier function compared to untreated physiological barrier repair. Transepidermal water loss (TEWL) measurements were performed. Before the start of the experiments, the skin tolerance of the cream was examined, revealing the non irritating characteristics of the ingredients and the absence of any contact allergic patch test reaction. In the ICD study, sodium lauryl sulfate (SLS) patches were applied to the forearms of young female volunteers. Consequently, it was observed that repeated cream application (14 days, 2x/day) significantly improved the TEWL of SLS-damaged skin, leading to a complete recovery on day 15. In the ACD study, disruption of skin barrier function was obtained by a nickel mediated contact allergy patch (CAP) test. The cream was then applied 2x/day for 4 consecutive days. Assessment of TEWL clearly showed that recovery of the disrupted skin significantly improved after cream application in comparison to untreated barrier repair. PMID- 11380544 TI - Fragrance contact dermatitis: a worldwide multicenter investigation (Part II). AB - The purpose of this study was to determine the frequency of responses to selected fragrance materials in patients who were fragrance sensitive. 178 patients were evaluated in 8 centers worldwide with a fragrance mix (FM) and 20 other fragrance materials. Reaction to the fragrance mixture (FM) occurred in 78.7% of the subjects. Substances reacting at a rate of 2% or higher included jasmine absolute, geranium oil bourbon, l-citronellol, spearmint oil, 1,3,4,6,7,8 hexahydro-4,6,6,7,8,8-hexamethylcyclopenta-gamma-2-benzopyran, omega-6 hexadecenlactone, dimethyltetrahydrobenzaldehyde (isomer mixture), and alpha amylcinnamaldehyde. These chemicals should be furthur evaluated to corroborate their allergenicity. We are constantly looking for new fragrance allergens to extend the diagnostic capability of the fragrance mix (FM). PMID- 11380545 TI - Mechanism of the antigen formation of carvone and related alpha, beta-unsaturated ketones. AB - In the present study, the mechanism for the antigen formation of alpha, beta unsaturated ketones was investigated. A series of analogues of carvone ((5R)-5 isopropenyl-2-methyl-2-cyclohexenone) with altered chemical reactivity and with retained overall structure or with retained reactivity and altered three dimensional structure were synthesized. These analogues were tested for cross reactivity in carvone-sensitized animals. Cross-reactivity was observed for analogue 3 ((5R)-5-isopropyl-2-methyl-2-cyclohexen-1-one). No cross-reactions were observed for analogues 1 ((2R,5R)-5-isopropenyl-2-methyl cyclohexanone) and 4 ((5R)-2,3-dimethyl-5-isopropenyl-2-cyclohexene-1-one). Both those compounds also failed to induce sensitization. These findings demonstrate that alpha, beta unsaturated ketones form antigens after a nucleophilic attack at the beta-carbon with soft nucleophiles such as thiol in cysteine and not with the formation of a Schiff's base after a nucleophilic attack at the carbonyl carbon with nitrogen nucleophiles. Furthermore, no cross-reactivity was observed between R- and S carvone indicating the importance of the 3-dimensional structure of haptens (and antigens) in T-cell recognition. The analogues were also tested for cross reactivity on patients allergic to carvone. The results from the animal study were confirmed. PMID- 11380546 TI - The suitability of hexyl cinnamic aldehyde as a calibrant for the murine local lymph node assay. AB - The murine local lymph node assay (LLNA) for the prospective identification of contact allergens assesses skin sensitization potential as a function of proliferative activity induced in lymph nodes draining the site of topical exposure to test chemical. This method has been endorsed recently as a stand alone test for the identification of contact allergens. We have now examined the suitability of hexyl cinnamic aldehyde (HCA), a recommended positive control for skin sensitization testing, as a calibrant for comparing the consistency of LLNA responses with time, and between laboratories, and thus for the routine assessment of assay reliability. Standard LLNAs were performed with CBA strain mice in 3 independent laboratories over a period of 8 years. Dose-response curves were used to derive mathematically the EC3 value (the estimated concentration of chemical necessary to cause a stimulation index (SI) of 3 compared with proliferation induced by concurrent vehicle controls). In each laboratory, 6 separate experiments were conducted using a single concentration of HCA (25%). Very similar stimulation indices were achieved, with mean values of 9.0, 6.5 and 6.6 recorded. A total of 10 dose-response experiments were performed independently in the 3 laboratories and these revealed that there was very little inter-laboratory, or temporal, variation in EC3 values. These data confirm that HCA responses in the LLNA are very stable and demonstrate that HCA provides a suitable calibrant for determining assay sensitivity and performance. PMID- 11380547 TI - Relevance and reproducibility of patch-test reactions to corticosteroids. AB - We present our patch test findings with dermatocorticosteroids during the period 1995-1999. We retrospectively studied the % of doubtful and positive reactions to each allergen in our corticosteroid series and assessed the relevance of these reactions. To assess reproducibility, we retested 15 patients with doubtful or positive reactions in 1999. The % of + or ++ reactions on D2-D3 and D7 ranged from 0.0% to 0.9% for most of the compounds tested. Hydrocortisone acetate 1% alc.abs./DMSO 50/50 showed a much higher % of positive reactions. Most of these reactions were not relevant and therefore probably due to the irritant nature of the vehicle. An overall reproducibility of 47.2% was obtained. 100.0% of the non reproducible patch test reactions were not relevant. Definite relevance was much higher in the reproducible + or ++ reactions than in the reproducible doubtful reactions: 100.0% and 18.2%, respectively. We conclude that the mean % of + and ++ reactions to the corticosteroid series on D2-D3 is 0.5%, with an overall reproducibility of 47.2%. Tixocortol pivalate and budesonide proved to be important markers for dermatocorticosteroid allergy. PMID- 11380548 TI - Clothing dermatitis from Naphthol AS. PMID- 11380549 TI - Allergic contact cheilitis from di-isostearyl malate in lipstick. PMID- 11380550 TI - IgE-mediated allergy to castor bean dust in a landscape gardener. PMID- 11380551 TI - Cold panniculitis--an unusual differential diagnosis from aluminium allergy in a patient hyposensitized with aluminium-precipitated antigen extract. PMID- 11380552 TI - Allergic contact dermatitis due to the beta-blocker befunolol in eyedrops, with cross-sensitivity to carteolol. PMID- 11380553 TI - Simultaneous photocontact sensitivity to ketoprofen and oxybenzone. PMID- 11380554 TI - Liquid-based cytology - an alternative international view. PMID- 11380555 TI - How predictive is a cervical smear suggesting invasive squamous cell carcinoma? AB - How predictive is a cervical smear suggesting invasive squamous cell carcinoma? Features have been described in severely dyskaryotic cervical smears that suggest frankly invasive or microinvasive squamous cell carcinoma. These are reported in three separate categories in our department. The aim of the current study was to assess the positive predictive value of these categories for invasive disease on histology. All smears reported in these categories over a five year period were correlated with the histology results. 527 smears were assessed. The positive predictive value of a smear suggesting frank invasion was 55.7% for all invasive squamous carcinomas and 40% for stage IB or above. Smears suspicious of invasion or microinvasion predicted invasive disease in 22.3% and 17.2%, respectively, most carcinomas being stage IA. Invasive squamous cell carcinoma may be predicted to a limited degree by cervical cytology especially when the smear suggests frank invasion. PMID- 11380556 TI - The preparation of additional smears from a cervical scrape: impact on the rate of detection of cervical neoplasia. AB - The preparation of additional smears from a cervical scrape: impact on the rate of detection of cervical neoplasia It has been known for some time that only a proportion of the cells on the smear-taking device is transferred to the slide. This can give rise to errors in reporting although the smear may have been taken correctly. This study was undertaken to identify a quick and simple method of improving the accuracy of the Papanicolaou test. A conventional smear and five additional smears were obtained from 62 women attending a Genito-Urinary Medicine clinic. The cell content of the conventional smears and the additional smears was compared. Dyskaryotic cells were detected both in the conventional smear and in the first and second additional smears from 22 women. Dyskaryotic cells were detected in the first and second additional smears only in five women. Thus, the conventional smear failed to detect biopsy-confirmed cervical abnormality in these women. A cell count of the first additional smear in the five cases where the conventional smear was negative showed that they contained, on average, 310 dyskaryotic cells. The preparation of one additional cervical smear per cervical scrape could significantly increase the accuracy of the cervical smear test by 11% (P=0.025, McNemar's test). PMID- 11380557 TI - Diagnosis of skeletal lymphoma and myeloma by radiology and fine needle aspiration cytology. AB - Diagnosis of skeletal lymphoma and myeloma by radiology and fine needle aspiration cytology From 1986 to 1998, all patients referred to Karolinska Hospital because of a skeletal destruction of unknown origin routinely underwent radiographic examination and fine needle aspiration cytology (FNAC). Among these, there were 83 patients with solitary lesions of the bone diagnosed and treated for myeloma (plasmacytoma) or non-Hodgkin's lymphoma. Review of the series showed that myeloma could not be distinguished radiographically from lymphoma. Nor could low and high grade lymphoma lesions be discriminated by radiographic appearance. The diagnostic utility of plain radiography in the two conditions seems to be confined to the mere detection of a destructive bone lesion and visual guidance for FNAC. The latter aspect, however, is crucial for the application of FNAC to bone lesions. Review of the cytologic specimens obtained by FNAC showed that they allowed a conclusive diagnosis in all 40 myeloma cases and in 41 of 43 lymphoma cases. In 32 of the 40 myeloma cases, the FNAC material could be used for immunocytochemistry, which disclosed kappa or lambda light chain restriction, corroborating the cytomorphological diagnosis. Thirty-eight lymphomas were characterized immunologically and in 35, a light chain restriction could be demonstrated. Our results show that the use of FNAC in the diagnosis of primary myeloma and lymphoma of bone obviates the need for other diagnostic modalities, including open biopsy. A combined approach based on radiology and FNAC, performed as an out-patient procedure, offers rapid and accurate diagnosis of myeloma and lymphoma among patients with radiographically unclassified destructive bone lesions. PMID- 11380558 TI - Immunophenotypic analysis of simultaneous specimens from different sites from the same patient with malignant lymphoma. AB - Immunophenotypic analysis of simultaneous specimens from different sites from the same patient with malignant lymphoma The assumption that immunophenotypic characteristics of different specimens obtained simultaneously from the same patient remain unchanged has rarely been evaluated. Using flow cytometry, we reviewed our experience of 29 patients with non Hodgkin's lymphoma (NHL). From these patients, 60 simultaneous specimens taken from the peripheral blood, bone marrow, effusions, fine needle aspirates from lymph nodes or cerebrospinal fluid were studied. In 26 out of 29 patients, the immunophenotype in the different specimens was identical. In one patient with unclassifiable low-grade B-NHL, immunophenotyping showed additionally a CD38 expression in the effusion which was not seen in the bone marrow. In one patient with mantle cell lymphoma, expression of CD10 in the lymph node was noted which was lacking in the peripheral blood. In the remaining patient with unclassifiable low-grade B-NHL, CD23 expression was noted in the lymph node but not in the peripheral blood. This retrospective study suggests that discordant antigen expression in samples from different body sites within the same patient is a rare event. PMID- 11380559 TI - Cytological diagnosis of soft tissue tumours. AB - Cytological diagnosis of soft tissue tumours The aims of this study were to determine the patterns of soft tissue tumours and also to try to assess the utility of fine needle aspiration cytology (FNAC) in diagnosing soft tissue tumours. Of 15 361 patients who visited the cytology diagnostic service of the Pathology Department, Medical Faculty, Addis Ababa University, 623 (4.1%) cases with a diagnosis of soft tissue tumours were retrieved from the department's records for the years 1991-96. Fifty-three soft tissue tumours (25 benign and 28 malignant tumours) with combined FNAC and surgical biopsy results were traced for cyto-histological correlations. Twenty-two out of 25 benign soft tissue tumours were correctly diagnosed, with three false cytologic diagnoses where one mesenchmal neoplasm, one haemangioma, and one haemorragic lesion were identified; and out of 28 malignant soft tissue, 23 were correctly diagnosed however, the five false cytological diagnoses were one soft tissue sarcoma, one dermatofibrosarcoma, one malignant mesenchymal neoplasm, one spindle cell neoplasm and one menechymal neoplasm. Thus, in this study a sensitivity and specificity of 88.5% and 81.5% respectively for the diagnosis of soft tissue tumours were reported. In conclusion, FNAC of soft tissue tumours is a fast, effective and reliable diagnostic tool that may help in categorizing soft tissue tumours into benign and malignant groups for clinical management. PMID- 11380560 TI - Cellular characteristics of nipple aspiration fluid during the menstrual cycle in healthy premenopausal women. AB - Cellular characteristics of nipple aspiration fluid during the menstrual cycle in healthy premenopausal women Fifteen healthy premenopausal female volunteers underwent weekly nipple aspiration of ductal fluid from both breasts during two menstrual cycles to investigate the variability of the cellular profile of the ductal fluid. Ductal fluid was successfully obtained using breast massage and nipple-areolar suction from 247/280 (89%) breasts. 83% of samples available for cytological analysis were cellular and 30% of cellular aspirates contained ductal epithelial cells identified using standard morphological criteria. No significant variation in cell number or cell type was identified during the menstrual cycle. All samples tested had an 'H' score of zero for oestrogen receptor. Seven out of 14 women expressed the proliferation marker Mcm-2 in the cells of at least one of the specimens, with no evidence of a menstrual cycle influence on expression. In conclusion, the cellular profile of breast ductal fluid did not vary consistently during the menstrual cycle, permitting future breast cancer screening studies incorporating serial nipple aspirations to be performed independent of the phase of the cycle. PMID- 11380562 TI - Cytological diagnosis of paucicellular variant of anaplastic carcinoma of thyroid: report of two cases. PMID- 11380561 TI - p53 protein expression and oestrogen and progesterone receptor status in invasive ductal breast carcinomas. AB - p53 protein expression and oestrogen and progesterone receptor status in invasive ductal breast carcinomas The p53 protein expression and oestrogen and progesterone receptors status was investigated in correlation to the grade of malignancy of primary breast carcinomas. Our material constituted imprints from surgical biopsies of 75 invasive ductal breast cancer cases. The p53 protein expression was investigated immunocytologically using the monoclonal antibody p53 DO-7 (DAKO). A biochemical DCC method was applied for the detection of oestrogen and progesterone receptors for all tumours. Fifty-one percent of breast cancer cases were p53 protein positive. A statistically significant association of p53 protein expression and high tumour grade was found (chi2=23.72, d.f.=2, P < 0.001). A statistically significant association was also found between oestrogen and progesterone receptor positive cases and the grade of malignancy (P < 0.001). A negative association between p53 protein expression and oestrogen (ER) and progesterone receptors (PgR) positivity was found. From our results it appears that it is possible to distinguish from grade II tumours two subgroups of cases, one with low malignancy potential and p53 (-), ER (+), PgR (+), and another subgroup with high malignancy potential and phenotype p53 (+), ER (-), PgR (-). The last subset of patients could actually benefit from adjuvant therapy. PMID- 11380563 TI - Maturational aspects of epilepsy mechanisms and consequences for the immature brain. PMID- 11380564 TI - Destruction of peripheral C-fibers does not alter subsequent vagus nerve stimulation-induced seizure suppression in rats. AB - PURPOSE: Early animal studies of the therapeutic mechanisms of vagus nerve stimulation (VNS) suggested that seizure suppression requires maximal activation of small, unmyelinated vagal C fibers. However, effective therapeutic stimulation parameters appear to be subthreshold for these fibers in humans, and there are no clinical reports of the autonomic side effects that would be expected if these fibers were maximally activated. We report here that selective destruction of C fibers with capsaicin does not affect VNS-induced seizure suppression in rats. METHODS: Rats were pretreated with capsaicin or vehicle in three injections over a 2-day period. A cuff electrode was later implanted on the left cervical vagus nerve. Two days after surgery, VNS was given to half of the capsaicin- and vehicle-treated rats. The remaining rats were connected to the stimulator but did not receive VNS. Thirty seconds after VNS onset, seizures were induced by pentylenetetrazol (PTZ), and seizure severity was measured. Two days later, the reciprocal VNS treatment was given, and PTZ-induced seizure severity was again measured. RESULTS: VNS effectively reduced seizure severity in both capsaicin- and vehicle-treated rats as compared with their non-VNS baselines. CONCLUSIONS: These results indicate that activation of vagal C fibers is not necessary for VNS induced seizure suppression. PMID- 11380565 TI - AWD 140-190: a potent anticonvulsant in the amygdala-kindling model of partial epilepsy. AB - PURPOSE: Evaluation of the effect of the new anticonvulsant drug, AWD 140-190 [4 (p-bromophenyl)-3-morpholino-1H-pyrrole-2-carboxylic acid methyl ester] on focally induced seizures and on epileptogenesis in the kindling model. METHODS: Effects of AWD 140-190 were studied in amygdala kindled rats after oral and intraperitoneal administration. In addition, the effect on kindling development was evaluated. In all experiments, behavioral changes in the rats in response to AWD 140-190 were monitored closely. RESULTS: AWD 140-190 exerted potent anticonvulsant activity against focal seizures. After intraperitoneal and oral administration in fully kindled rats, the substance dose-dependently increased the threshold for induction of afterdischarges starting at 15 mg/kg. AWD 140-190 only weakly influenced the seizure severity of the animals after stimulation at the elevated afterdischarge threshold current. No adverse effects were observed up to 30 mg/kg after intraperitoneal and oral administration in the open field and in the rotarod test. No differences were found between kindled and nonkindled rats when comparing neurotoxicity of AWD 140-190. Prolonged treatment with AWD 140-190 during kindling acquisition did not prevent kindling, but significantly retarded the development of fully kindled seizures during the treatment. CONCLUSIONS: This study demonstrates that AWD 140-190 has anticonvulsant effects in the amygdala kindling model in rats, suggesting that the substance is particularly effective against partial seizures. AWD 140-190 is orally active and devoid of neurotoxic effects in anticonvulsant doses, thus indicating that this compound has potential for antiepileptic therapy. AWD 140-190 retards the kindling development during the treatment. This effect could be explained by the acute anticonvulsant effect of the substance. PMID- 11380566 TI - Interaction of the novel anticonvulsant, BIA 2-093, with voltage-gated sodium channels: comparison with carbamazepine. AB - PURPOSE: BIA 2-093 [(S)-(-)-10-acetoxy-10,11-dihydro-5H-dibenz/b,f/azepine-5 carboxamide] is endowed with an anticonvulsant potency similar to that of carbamazepine (CBZ), but produces less cognitive and motor impairment. This study evaluated whether voltage-gated sodium channels (VGSCs) are a primary locus for the action of BIA 2-093. METHODS: We used the whole-cell voltage-clamp technique in the mouse neuroblastoma cell line N1E-115 to investigate the effects of BIA 2 093 and CBZ on VGSCs, displacement of [3H]-batrachotoxinin A 20-alpha-benzoate ([3H]-BTX), and [3H]-saxitoxin to define their relative potency to bind to rat brain sodium channels, and inhibition of uptake of 22Na by rat brain cortical synaptosomes stimulated by veratridine as a measure of sodium entry. RESULTS: The inhibitory potencies of BIA 2-093 and CBZ increased as the holding potential was made less negative (-100, -90, -80, and -70 mV) with median inhibitory concentration (IC50) values (in microM) of, respectively, 4,337, 618, 238, and 139 for BIA 2-093, and 1,506, 594, 194, and 101 for CBZ. BIA 2-093 displayed a similar potency in displacing [3H]-BTX (IC50 values, 222 vs. 361 microM; p > 0.05) and inhibiting the uptake of 22Na (IC50 values, 36 vs. 138 microM; p > 0.05). Both drugs failed to displace [3H]-saxitoxin in concentrations up to 300 microM. CONCLUSIONS: BIA 2-093, like CBZ, inhibits sodium currents in a voltage dependent way by an interaction predominantly with the inactivated state of the channel and interacts with neurotoxin receptor site 2, but not with receptor site 1. BIA 2-093 displayed a potency blocking VGSCs similar to that of CBZ. PMID- 11380567 TI - Vigabatrin visual toxicity: evolution and dose dependence. AB - PURPOSE: To investigate the prevalence and prognosis of visual field defects (VFDs) in epilepsy patients with and without vigabatrin (VGB) treatment; to investigate the possible relationship between VFDs and cumulative VGB dose, and to characterise the evolution of VFDs. METHODS: A cohort of 155 presurgical candidates who had undergone full-field Goldmann perimetry (GP) was studied, 99 (64%) of whom had been treated with VGB. All GPs were reevaluated in 1998 by one experienced examiner, blinded to medication. Duration of treatment and total VGB dose were related to perimetric results. RESULTS: Twenty-five (16%) of the 155 patients had VFDs: Nineteen (19%) of the 99 VGB-treated patients, and six (11%) of the 56 patients unexposed to VGB. VGB-treated patients with VFDs had been treated significantly longer than those without VFDs. Cumulative VGB dose could be calculated for 84 patients. The prevalence of VFDs increased significantly with increasing total VGB-dose, from 4% in the 51 patients who had been exposed to C (exon 1) and Asp (A) 96 Asp (G) (exon 4). In Nancy, the T + 70-->C polymorphism was associated with fat mass adjusted for BMI in women (P = 0.025). The genotype and allele frequencies of the Ser (T) 343 Ser (C) polymorphism (exon 9) were significantly different between normal and overweight women, with the T allele being more frequent in the overweight group (T frequency in Nancy, 0.82; in Nancy + Paris, 0.79) than in the normal weight group (0.69; P = 0.017 vs. Nancy overweight, P = 0.003 vs. Nancy + Paris overweight). In women from Nancy, fat mass adjusted for BMI was significantly associated with this polymorphism (P = 0.01). The overweight women carrying the C allele of this polymorphism lost more weight in response to low calorie diet than the non carriers (P = 0.006). CONCLUSIONS: In women, genetic variations at the LEPR gene level are associated with overweight and fat mass in a cross sectional study and with response to low calorie diet in an intervention study. These results indicate that variations at the leptin receptor locus are associated with common obesity phenotypes and are a part of the polygenic influences on the response to nutritional environment. PMID- 11380592 TI - Assessment of beta-cell function during the oral glucose tolerance test by a minimal model of insulin secretion. AB - OBJECTIVE: To characterise the performance of beta-cell during a standard oral glucose tolerance test (OGTT). DESIGN: Fifty-six subjects were studied. A minimal analogic model of beta-cell secretion during the OGTT was applied to all OGTTs (see below). The amount of insulin secreted over 120' in response to oral glucose (OGTT-ISR; Insulin Units 120'-1 m-2 BSA) and an index of beta-cell secretory 'force' (beta-Index; pmol.min-2.m-2 BSA) were computed with the aid of the model. In protocol A, 10 healthy subjects underwent two repeat 75 g OGTT with frequent (every 10'-15') blood sampling for glucose and C-peptide to test the reproducibility of OGTT-ISR and beta-Index with a complete or a reduced data set. In protocol B, 7 healthy subjects underwent three OGTTs (50, 100 or 150 g), to test the stability of the beta-Index under different glucose loads. In protocol C, 29 subjects (15 with normal glucose tolerance, 7 with impaired glucose tolerance and 7 with newly diagnosed type 2 diabetes) underwent two repeat 75 g OGTT with reduced (every 30' for 120') blood sampling to compare the reproducibility and the discriminant ratio (DR) of OGTT-ISR and beta-index with the insulinogenic index (IG-Index: Delta Insulin 30' - Basal/Delta Glucose 30' - Basal). In protocol D, 20 subjects (14 with normal glucose tolerance, 5 with impaired glucose tolerance and 1 with newly-diagnosed type 2 diabetes) underwent a 75 g OGTT and an intravenous glucose tolerance test (IVGTT) on separate days to explore the relationships between acute (0'-10') insulin response (AIR) during the IVGTT and beta-index and OGTT-ISR during the OGTT. RESULTS: In all protocols, the minimal analogic model of C-peptide secretion achieved a reasonable fit of the experimental data. In protocol A, a good reproducibility of both beta-index and OGTT-ISR was observed with both complete and reduced (every 30') data sets. In protocol B, increasing the oral glucose load caused progressive increases in OGTT-ISR (from 2.63 +/- 0.70 to 5.11 +/- 0.91 Units.120'-1.m-2 BSA; P < 0.01), but the beta-index stayed the same (4.14 +/- 0.35 vs. 4.29 +/- 0.30 vs. 4.30 +/- 0.33 pmol.min-2.m-2 BSA). In protocol C, both OGTT-ISR and beta-index had lower day-to-day CVs (17.6 +/- 2.2 and 12.4 +/- 2.4%, respectively) and higher DRs (2.57 and 1.74, respectively) than the IG-index (CV: 35.5 +/- 6.3%; DR: 0.934). OGTT-ISR was positively correlated to BMI (P < 0.03), whereas beta-index was inversely related to both fasting and 2 h plasma glucose (P < 0.01 for both). In protocol D, beta-index, but not OGTT-ISR, was significantly correlated to AIR (r = 0.542, P < 0.02). CONCLUSIONS: Analogically modelling beta-cell function during the OGTT provides a simple, useful tool for the physiological assessment of beta cell function. PMID- 11380593 TI - Glutathione and alpha-lipoate in diabetic rats: nerve function, blood flow and oxidative state. AB - BACKGROUND: Increased oxidative stress is considered to be a causal factor in the development of diabetic complications, among which peripheral neuropathy. The pathophysiology of nerve dysfunction in diabetes has been explained both by reduced endoneurial microcirculation and alterations in endoneurial metabolism. It is unclear whether antioxidants primarily improve nerve blood flow or normalise systemic or endoneurial oxidative metabolism. Therefore, we evaluated the effects of the antioxidants glutathione and alpha-lipoic acid on both nerve microcirculation and the antioxidative capacity and lipid peroxidation in experimentally diabetic rats. MATERIALS AND METHODS: Streptozotocin-diabetic rats were treated with different doses of alpha-lipoic acid, reduced glutathione or placebo, and were compared with nondiabetic controls. We measured systemic and endoneurial antioxidants, malondialdehyde and whole blood hydrogen peroxide. Furthermore, we evaluated sciatic and tibial motor and sensory nerve conduction velocity, caudal nerve conduction velocity, and assessed sciatic nerve blood flow and vascular resistance by Laser-Doppler flowmetry. RESULTS: We observed a rise in erythrocyte glutathione by 27 % (P < 0.05), and a trend towards decreased plasma malondialdehyde in alpha-lipoic acid, but not in glutathione-treated animals in comparison with the placebo group. Simultaneously, sciatic nerve blood flow and vascular resistance were improved by daily alpha-lipoic acid administration by 38% (P < 0.05). Peripheral nerve conduction velocity and endoneurial glutathione were not significantly influenced by antioxidant treatment. CONCLUSIONS: Only minor beneficial effects of alpha-lipoic acid on nerve blood flow and oxidative state occur at the given doses; these effects were insufficient to improve nerve conduction deficits. PMID- 11380595 TI - Helicobacter pylori-related diseases. PMID- 11380594 TI - Fecal elastase-1 is decreased in villous atrophy regardless of the underlying disease. AB - BACKGROUND: Fecal elastase-1 (E1) is a sensitive and reliable test in the assessment of exocrine pancreatic function in cystic fibrosis (CF). In patients with celiac disease (CD), different E1 values have been reported. E1 levels in other malabsorption conditions are unknown. Therefore, the aim of the present study was to evaluate E1 concentrations in various malabsorption syndromes. MATERIAL AND METHODS: The study was carried out in 54 patients, selected from patients referred with suspicion of CF, who had been diagnosed as celiac disease (CD; n = 16), secondary malabsorption syndrome (SMS, giardiasis- or cow milk related enteropathy; n = 18) and food allergy (FA; n = 20). 70 age-matched healthy children (HC) and 131 cystic fibrosis (CF) patients served as control groups. In CD and SMS patients, a gluten-free diet was introduced. In addition, SMS patients were treated appropriately to underlying disease. In all subjects, E1 concentrations were measured. In CD and SMS patients, E1 concentrations were repeatedly measured after one year of the treatment. RESULTS: With a cut-off level of 200 microg g-1, abnormal E1 concentrations were found in 87.2% of the CF group and in 56.2% and 50.0% of the CD and SMS subgroups, respectively. In none of FA patients, were E1 values below the normal range. After mucosal recovery, E1 concentrations in patients with CD and SMS increased, suggesting that villous atrophy can diminish exocrine pancreatic secretion. In 18 out of 19 CD and SMS patients with abnormal E1 concentrations, monitored for at least 12 months of a gluten-free diet, abnormal E1 concentrations increased above the cut-off value to normal range. Two out of the 54 referred patients were finally diagnosed as having CF, one with stable low E1 levels and the second with finally normal values. CONCLUSIONS: The exocrine pancreatic function is decreased in villous atrophy regardless of underlying disease. The specificity of the fecal elastase-1 test in the differentiation between 'primary' exocrine pancreatic insufficiency and intestinal malabsorption with mucosal atrophy is low. After mucosal regeneration, fecal elastase-1 specificity is high. PMID- 11380596 TI - Type I collagen markers in cord serum of appropriate vs. small for gestational age infants born during the second half of pregnancy. AB - BACKGROUND: The serum concentration of the N-terminal propeptide of type I procollagen (PINP) reflects the synthesis rate of type I collagen, whereas the corresponding C-terminal telopeptide (ICTP) mirrors its degradation. DESIGN: PINP and ICTP were measured in a total of 690 cord serum samples from 592 appropriate for-gestational-age (AGA) infants and 98 smal-for-gestational-age (SGA) infants. These markers were compared between AGA and SGA infants of different gestational ages, ranging from 23 to 41 weeks, and birth weights, from 620 to 4555 g. RESULTS: Both PINP and ICTP levels were very high in the preterm AGA infants and declined significantly with advancing gestational age, paralleling the shape of the fetal growth velocity curve. Regardless of the quite large interindividual variations observed in these markers, PINP was significantly lower in both the preterm and term AGA infants than in the SGA infants. This was also the case for ICTP in the preterm infants of gestational age less than 36 weeks. In stepwise multiple regression analyses, gestational age, being either AGA or SGA and head circumference were significant factors to explain the levels of PINP and ICTP. The levels of PINP and ICTP were correlated with each other highly significantly in both the AGA and SGA infants (rs = 0.700 and 0.692, respectively; P < 0.001 in both). CONCLUSIONS: The levels of type I collagen markers seem to follow closely the shape of the fetal growth velocity curve during different stages of gestation. However, because of the large interindividual variations observed, further studies are needed before the significance of these markers for the assessment of normal and abnormal fetal growth can be established. PMID- 11380597 TI - Bone turnover markers and sex hormones in men with idiopathic osteoporosis. AB - BACKGROUND: In contrast to osteoporosis in postmenopausal women, osteoporosis in men has received much less attention. PATIENTS AND METHODS: We determined various biochemical parameters of bone metabolism and sex hormones in 31 men with idiopathic osteoporosis and 35 age matched control subjects. RESULTS: In the men with osteoporosis, a significantly increased urinary excretion of deoxypyridinoline (5.3 +/- 0.2 vs. 4.6 +/- 0.2 nmol mmol-1 creatinine; P = 0.033) in addition to increased serum levels of the c-terminal telopeptide of type I collagen (2677 +/- 230 vs. 2058 +/- 153 pmol; P = 0.037) were found. While parameters of bone formation were not significantly different in the patients and controls, serum bone sialoprotein levels were significantly decreased in the patients (3.7 +/- 0.8 vs. 12.4 +/- 4.0 ng mL-1; P = 0.021). Moreover, in men with idiopathic osteoporosis, lower levels of estradiol (91.3 +/- 5.8 vs. 114.6 +/- 7.8 pmol L-1; P = 0.044), higher levels of sex hormone binding globulin (31.5 +/- 3.1 vs. 24.2 +/- 1.4 nmol L-1; P = 0.034) and a decreased free androgen index (42.6 +/- 5.2 vs. 56.4 +/- 5.9; P = 0.016) were seen. Serum estradiol levels correlated negatively with several parameters of bone resorption. CONCLUSIONS: In men with idiopathic osteoporosis, bone resorption is increased and exceeds bone formation. The excessive bone resorption seen in idiopathic male osteoporosis may be due to decreased estradiol levels and low levels of bioavailable testosterone. PMID- 11380598 TI - N-acetyl-L-cysteine exerts direct anti-aggregating effect on human platelets. AB - BACKGROUND: N-acetyl-L-cysteine, a thiol compound, has been shown to potentiate the inhibition of platelet aggregation exerted by organic nitrates and to increase the anti-aggregating effect of L-arginine, which promotes endogenous synthesis of nitric oxide (NO) acting as substrate of platelet constitutive nitric oxide synthase (NOS). It is not known whether this thiol can exert direct effects on platelet aggregability. MATERIALS AND METHODS: 14 healthy male volunteers provided platelet samples to investigate whether N-acetyl-L-cysteine directly influences platelet function and intraplatelet levels of 3',5' cyclic guanosine monophosphate (cGMP), which represents the second messenger involved in NO-induced antiaggregation. Some experiments were repeated in the presence of NOS inhibitor NG-monomethyl-L-arginine (L-NMMA), of nitric oxide-sensitive guanylyl cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ), of the selective cGMP phosphodiesterase inhibitor zaprinast and of calcium ionophores (A23187, ionomycin). RESULTS: N-acetyl-L-cysteine at 3000-6000 micromol L-1 decreases the responses of human platelets both in platelet-rich plasma (aggregation induced by adenosine 5-diphosphate) and in whole blood (aggregation induced by collagen). The anti-aggregating effect was prevented by preincubation with L-NMMA and guanylyl cyclase inhibitor ODQ. In resting platelets, N-acetyl-L cysteine increased the levels of cGMP starting from a concentration of 3000 micromol L-1. Permeabilized platelets exhibited an increased sensitivity to the anti-aggregating effect of N-acetyl-L-cysteine. Also, cGMP phosphodiesterase inhibition or the increase in calcium availability, enhanced N-acetyl-L-cysteine effects on platelets. CONCLUSION: N-acetyl-L-cysteine exerts direct anti aggregating effects through an increased bioavailability of platelet nitric oxide. PMID- 11380599 TI - Effect of nitric oxide inhibition on nasal airway resistance after nasal allergen challenge in allergic rhinitis. AB - BACKGROUND: Nitric oxide has been detected by chemiluminescence in the lumen of nasal airway, which is increased in nasal breathing in patients with seasonal rhinitis during a chronic exposure. The purpose of this study was to determinate the effect of a NO-synthase inhibitor NGL-arginine methyl ester (L-NAME) on nasal airway resistance (NAR) in patients with seasonal allergic rhinitis after an acute challenge to the allergen. METHODS: Nitric oxide levels in the nose were measured by the chemiluminescence method in nine non-atopic volunteers and in seven patients with seasonal rhinitis at rest and after an acute challenge with the allergen. NAR were measured by active anterior rhinomanometry. RESULTS: Basal nasal NO concentration in allergic rhinitis was 496.5 +/- 151.4 parts per billion (ppb). (n = 7) and it was not significantly different from levels found in the control group: 458.4 +/- 105.9 ppb (n = 9). The topical administration of L-NAME in allergic rhinitis reduced the NO concentration (338.6 +/- 99.3 ppb, P < 0.001; n = 7). In the rhinitic patients the challenge with the allergen did not modify the nasal NO levels (504.5 +/- 138.5 ppb). The application of the allergen after the pretreatment with placebo caused a significant increase in NAR (from 0.32 +/- 0.11 Pa s cm-3 to 1.01 +/- 0.12 Pa s cm-3, P < 0.001; n = 7). Pre-treatment with L-NAME did not prevent the increase in NAR induced by allergen challenge (from 0.36 +/- 0.15 Pa s cm-3 to 1.06 +/- 0.26 Pa s cm-3). CONCLUSIONS: The results indicate that nasal administration of a NOS inhibitor L-NAME, at doses capable of decreasing nasal NO levels, has no effect on NAR and it does not prevent the NAR increase induced by an acute challenge with allergen in subjects with seasonal rhinitis. PMID- 11380600 TI - Study of cord blood natural killer cell suppressor activity. AB - We tested the immunosuppressive effect of cord blood (CB) natural killer (NK) cells using highly purified CB NK cells in mixed lymphocyte cultures (MLC) containing autologous CB T cells as responders. Control cultures were done without NK cells. Our findings revealed that CB NK cells induced a dose-dependent inhibition of T lymphocyte proliferation as evidenced by decreased 3H-thymidine incorporation in MLC. The T cell alloproliferation was significantly decreased in the presence of an NK cell to responder cell ratio of 0.1, 0.2 or 0.4 compared with control cultures done without NK cells (p=0.02, 0.003 and 0.0002, respectively). T lymphocyte inhibition was also achieved using irradiated CB NK cells and still demonstrable on addition of disparate CB NK and T cells to the MLC. In agreement with previous reports, adult blood NK cells inhibited the alloreactive T cells in the MLC using adult T lymphocytes as responders. Compared to control cultures done without NK cells, statistically significant inhibition of 3H-thymidine incorporation in MLC was observed at a ratio of NK cells to responder cells ratio of 0.2 or 0.4 (p=0.02). To investigate the mechanism whereby CB NK cells can interfere with the development of alloreactive T cells in MLC, we measured the tumour necrosis factor-alpha (TNF-alpha) concentrations in MLC supernatants using NK cell-depleted or unseparated CB mononuclear cells (MNC) as responders. The results revealed significantly high levels of TNF-alpha in the absence of NK cells (p=0.007). We conclude that CB NK cells suppress alloreactive T lymphocytes as do their counterparts in adult blood. However, the high NK to T cell ratio in CB could contribute to a more marked suppressive potential compared to that in adult blood. The mechanism of NK-mediated inhibition is likely related to disruption of the TNF-alpha pathway of T-lymphocyte activation. PMID- 11380601 TI - Changes in murine bone marrow macrophages and erythroid burst-forming cells following the intravenous injection of liposome-encapsulated dichloromethylene diphosphonate (Cl2MDP). AB - In order to explore the effect on bone marrow macrophages of liposome encapsulated dichloromethylene diphosphonate (Cl2MDP), mice were injected intravenously with a preparation of such liposomes at a dose known to deplete spleen and liver macrophages. Two days later, the macrophages in the marrow of the femoral bones were quantified by flow cytometry using a macrophage-specific monoclonal antibody (F4/80), and their ultrastructure and phagocytic activity towards zymosan particles was assessed. To determine the effect on erythropoiesis of liposome-encapsulated Cl2MDP-induced changes in bone marrow macrophages, red blood cell parameters and the formation of erythroid burst-forming unit (BFU-E) derived colonies in vitro were evaluated. In mice injected with liposome encapsulated Cl2MDP, there was a 54% and 67% decrease in the total number of bone marrow macrophages as compared to uninjected controls and mice treated with empty liposomes, respectively. Moreover, residual macrophages showed an abnormal ultrastructure, with reduced numbers of crystalloid inclusions and increased numbers of large myelin figures. However, the phagocytic activity of these cells was unimpaired or slightly enhanced. In mice injected with liposome-encapsulated Cl2MDP there was an approximately 60% decrease in the percentage and total number of circulating reticulocytes and a 54% reduction in the BFU-E number, demonstrating deregulation of erythropoiesis under conditions of macrophage loss and impairment. The results suggest that mice treated with liposome-encapsulated Cl2MDP are a model for studying the role of macrophages in erythropoiesis. PMID- 11380603 TI - Decrease of bone marrow angiogenesis in myeloma patients achieving a remission after chemotherapy. AB - The impact of angiogenesis is well known for the growth and viability of solid tumors. Fewer studies have been published relating angiogenesis to clinical or pathological parameters in hematological malignancies. In this report, we have estimated the bone marrow microvessel density (MVD) before and after conventional dose or high-dose chemotherapy with autologous stem cell transplantation. Immunohistochemical CD34-stained paraffin-embedded bone marrow biopsies of 21 patients with stage III multiple myeloma were studied. Microvessels were counted at 400x magnification, and the mean number of vessels per area in each sample was noted as the MVD. The median MVD of all patients was 53.1 vessels/mm2 (range 15.5 174.7 vessels/mm2) before treatment and 29.3 vessels/mm2 (range 0-221.1 vessels/mm2) after chemotherapy. The post-treatment MVD in the two groups of patients with and without remission was significantly different (p=0.001), whereas the pretreatment MVD was not. Responders but not nonresponders showed a significant decrease of MVD after therapy in comparison to their pretreatment levels. The progression-free survival in patients who achieved a reduction in MVD after chemotherapy was significantly longer than in patients without a decrease in MVD (P=0.006). Furthermore, we compared the MVD of patients after achievement of a remission to MVD of 15 untreated stage I myeloma patients. The MVD of patients in remission was not statistically different from the MVD in stage I myeloma. These results underscore the impact of angiogenesis in myeloma and give the first report that effective chemotherapy is accompanied by a significant decrease in bone marrow angiogenesis in this disease. PMID- 11380602 TI - Bone marrow stromal dysfunction in mice administered cytosine arabinoside. AB - OBJECTIVE: The aim of the study was to investigate ex-vivo the bone marrow (BM) stroma of mice under conditions of low- and high-dose cytosine arabinoside (Ara C), a cycle-specific drug (S-phase) and to assess possible stromal damage, apart from the killing of hematopoietic cells. Stroma consists of mesenchymal elements generally not in the cell cycle; therefore it could not be a target for the killing effect of Ara- C. MATERIALS AND METHODS: The stromal function was studied by the following: the incidence of stromal stem cells, i.e. CFU-F; formation of stromal layers under growth conditions of long-term culture (LTC) followed by irradiation and overlayering of test cells in contact and non-contact co cultures; subsequent culture of the test cells in a semi-solid medium to assay the incidence of hyperproliferative potential cells (HPPC); production of GM-CSF, IL-3, IL-4, IL-6 and IFNgamma in the conditioned medium (CM) of confluent stromal layers. All tests and assays were carried out on BM specimens, 1-4 d after Ara-C administration and on controls. RESULTS: Low-dose Ara-C induces a marked decrease of CFU-F, compensated by cycle induction of pre-CFU-F, young-type stromal stem cells. High-dose Ara-C causes a CFU-F decrease to almost zero level. The time length to layer confluency is normal after low-dose Ara-C ( approximately 10 d) and prolonged after a high dose ( approximately 30 d). The confluent layers from mice receiving low- or high-dose Ara-C support hematopoiesis adequately. Among the growth factors and cytokines assayed, only IL-6 is detected in CM layers. IL 6 decreases after a low dose of Ara-C and increases after a high dose. The cause of IL-6 fluctuations is yet to be investigated. It is, however, evident that IL-6 is not an essential factor in support of hematopoiesis. CONCLUSIONS: Taken together, the current study in mice indicates that Ara-C administration, in particular a high dose, induces bone marrow stromal damage and/or disfunction. The long period of time to reach layer confluency after a high Ara-C dose might reflect the in-vivo situation of slow stromal regeneration. PMID- 11380604 TI - Decreased amount of mpl and reduced expression of glycoprotein IIb/IIIa and glycoprotein Ib on platelets from patients with refractory anemia: analysis by a non-isotopic quantitative ligand binding assay and immunofluorescence. AB - Using a non-isotopic ligand binding assay and immunofluorescence, we examined the amount of mpl, glycoprotein IIb/IIIa (gpIIb/IIIa) and glycoprotein Ib (gpIb) on platelets from healthy volunteers and patients with refractory anemia (RA). For the analysis of mpl expression, we applied both a non-isotopic ligand binding assay and immunofluorescence using anti-mpl monoclonal antibody, and compared the results from both methods. The non-isotopic ligand binding assay has been developed in our laboratory and is suitable for the quantitative analysis of a small amount of cytokine receptors such as mpl on platelets. In platelets from patients with RA, the amount of mpl expressed by the D value was 0.05+/-0.03 (mean+/-standard deviation), and was significantly lower than that in healthy volunteers (0.15+/-0.05, p<0.0001). The mean fluorescence intensities (MFI) of gpIIb/IIIa and gpIb on platelets from RA patients were 28.8+/-8.8 and 20.8+/-7.7, respectively, and were significantly lower than those on normal subjects (93.2+/ 22.6 and 67.4+/-9.1, p<0.0001 and p<0.0001, respectively). There was a good correlation between the amount of mpl and the MFI of gpIIb/IIIa (p=0.794, p<0.0001) or gpIb (p=0.774, p<0.0001), and between those of gpIIb/IIIa and gpIb (p=0.728, p<0.0001). We demonstrated a decreased amount of mpl as well as a reduced expression of gpIIb/IIIa and gpIb on platelets from RA patients. PMID- 11380605 TI - Laboratory diagnosis of heparin-induced thrombocytopenia: advantages of a functional flow cytometric test in comparison to the heparin-induced platelet activation test. AB - Nearly one third of patients with heparin-induced thrombocytopenia (HIT) will progress to overt thrombosis. Owing to the severity of HIT, a reliable prompt diagnosis is mandatory. In this study 248 consecutive samples from patients referred to our laboratory for HIT diagnosis and 97 specimens from normal controls were prospectively evaluated in parallel using the heparin-induced platelet aggregation (HIPA) test and a flow cytometric (FC) test. The HIPA test resulted in 214 negative, 17 indeterminate and 17 positive samples of patients. The FC method detects activated platelets induced by heparin-immune complexes using the highly sensitive recombinant probe annexin V and pooled platelets from multiple donors. The criteria for positive FC test results included an increase in platelet activiation of at least 11% at 0.3 IU/mL heparin concentration in the tube, and a ratio of more than 1.5 between platelet activation at 0.3 and 200 IU/mL heparin. According to the cut-off level 17 patients who showed indeteminate HIPA test results had 14 negative and 3 indeterminate corresponding FC test results. Only one of these patients (HIPA test indeterminate, FC test indeterminate) had no other obvious medical cause for thrombocytopenia than HIT. Infections or inflammations did not show any association with the FC test results, whereas thromboembolic events displayed a significant patelet activation at pharmacological heparin concentration. Therefore the FC test is associated to the complications of HIT. In conclusion, the FC test, which is fast and practical, showed a good agreement with the HIPA test and may be an accurate and useful test for HIT. PMID- 11380606 TI - Doxorubicin and a butyric acid derivative effectively reduce levels of BCL-2 protein in the cells of chronic lymphocytic leukemia patient. AB - B-chronic lymphocytic leukemia (B-CLL) is a disease caused primarily by defects in the apoptosis mechanism. AN-9, a butyric acid (BA) derivative, is a potent differentiating and an anti-cancer drug that induces apoptosis in HL-60 cells. Herein we show the affect of AN-9, alone and in combination with doxorubicin, on cell cultures from B-CLL patients. Cells from 17 patients were cultured and tested for viability, apoptosis, bcl-2 and bax protein expression. Exposure of B CLL cell cultures to AN-9 was accompanied by apoptosis and a marked viability loss (up to 46%, p=0.0017). AN-9 reduced up to 51% (p=0.0017) the levels of bcl-2 in 57% of the cultures that express bcl-2. The combination of low concentrations of AN-9 and doxorubicin more than additively enhanced apoptosis and reduced bcl-2 levels in B-CLL cultures which were resistant to AN-9. AN-9 enhanced bax expression up to 58%(p=0.008) in cultures from 53% of the patients, but had no effect on bax levels when combined with doxorubicin. In conclusion, AN-9 alone reduced bcl-2 and enhanced bax expression in cultures from B-CLL patients, and the reduction of bcl-2 levels in combination with doxorubicin was greater than additive. These results may be beneficial in possible future combination therapy with AN-9 in B-CLL. PMID- 11380607 TI - Haematopoietic progenitor cells from the common marmoset as targets of gene transduction by retroviral and adenoviral vectors. AB - To establish a new non-human primate model for human cytokine and gene therapy, we characterized lymphocytes and haematopoietic progenitor cells of the small New World monkey, the common marmoset. We first assessed the reactions of marmoset bone marrow (BM) and peripheral blood (PB) cells to mouse anti-human monoclonal antibodies (mAbs) for the purpose of isolating marmoset lymphocytes and haematopoietic progenitor cells. Both cell fractions stained with CD4 and CD8 mAbs were identified as lymphocytes by cell proliferation assay and morphological examination. Myeloid-specific mAbs such as CD14 and CD33 did not react with marmoset BM and PB cells. No available CD34 and c-kit mAbs could be used to purify the marmoset haematopoietic progenitor cells. Furthermore, we studied the in vitro transduction of the bacterial beta-galactosidase (LacZ) gene into CFU-GM derived from marmoset BM using retroviral and adenoviral vectors. The transduction efficiency was increased by using a mixed culture system consisting of marmoset BM stromal cells and retroviral producer cells. It was also possible to transduce LacZ gene into marmoset haematopoietic progenitor cells with adenoviral vectors as well as retroviral vectors. The percentage of adenovirally transduced LacZ-positive clusters was 15% at day 4 (multiplicity of infection=200), but only 1-2% at day 14. The differential use of viral vector systems is to be recommended in targeting different diseases. Our results suggested that marmoset BM progenitor cells were available to examine the transduction efficiency of various viral vectors in vitro. PMID- 11380608 TI - Cytogenetic and immunophenotypic evidence of independent clonal origins of concomitant chronic lymphocytic leukaemia and acute myeloid leukaemia. PMID- 11380609 TI - RNA as a tumor vaccine: a review of the literature. AB - Many approaches have been attempted to harness the host immune system to act against malignant tumors. These have included animal and clinical trials with agents to non-specifically boost immunity, factors to augment specific immunity, transfer of lymphokine-activated killer cells and transfer of expanded populations of tumor-infiltrating lymphocytes. Therapeutic vaccination strategies have been employed using tumor extracts, purified tumor antigens, recombinant peptide tumor antigens and specific DNA sequences coding for a tumor antigen (genetic vaccination) both through direct administration to the host and by administration of antigen presenting cells exposed to these materials ex vivo. Recently, the use of RNA has been proposed for use in tumor vaccination protocols. The use of RNA has several potential advantages. Since total cellular RNA or mRNA can be utilized, it is not necessary to know the molecular nature of the putative tumor antigen(s). RNA can be effectively amplified; thus, unlike tumor-extract vaccines, only a small amount of tumor is needed to prepare the material for vaccination. Also, unlike DNA-based vaccines, there is little danger of incorporation of RNA sequences into the host genome. The possible utility of RNA-based vaccines for tumor immunotherapy should be further explored to determine whether such approaches are clinically useful. PMID- 11380610 TI - The molecular determinants of sunburn cell formation. AB - Sunburn cell (SBC) formation in the epidermis is a characteristic consequence of ultraviolet radiation (UVR) exposure at doses around or above the minimum erythema dose. SBC have been identified morphologically and biologically as keratinocytes undergoing apoptosis. There is evidence that SBC formation is a protective mechanism to eliminate cells at risk of malignant transformation. The level of DNA photodamage is a major determinant of SBC induction by a process controlled by the tumor suppressor gene p53. However, extra-nuclear events also contribute to SBC formation, such as the activation of death receptors including CD95/Fas. UVR triggers death receptors either by direct activation of these surface molecules or by inducing the release of their ligands such as CD95 ligand or tumor necrosis factor. Oxidative stress also appears to be involved, probably via mitochondrial pathways, resulting in the release of cytochrome C. Pathways which modify SBC formation are now extensively studied given the importance of apoptosis in eliminating irreparably damaged cells. A greater understanding of the mechanisms that induce and prevent UVR-induced apoptosis will contribute to our understanding of mechanisms relevant in genomic integrity. PMID- 11380611 TI - Successful induction of immune responses against mutant ras in melanoma patients using intradermal injection of peptides and GM-CSF as adjuvant. AB - The rapidly increasing incidence and mortality rate of malignant melanoma, together with the lack of efficient treatment of the late stages, makes it a serious threat to public health. Innovative new treatments are needed. The proteins of the ras-family of proto-oncogenes, functioning as relay switches for signalling pathways between cell surface and nucleus, are involved in cell proliferation, differentiation, apoptosis and transformation. If over-expressed or mutated they can induce and/or maintain a transformed state of a cell. Codon 61 mutations of N-ras seem to be involved in melanoma development on sun exposed sites. In order to induce an immune response towards mutated N-ras proteins we performed a phase 1 feasibility study. Ten melanoma patients were immunized intradermally 6 times with N-ras peptides (residue 49-73) with 4 codon 61 mutations using GM-CSF as adjuvant. HLA typing was not used as an inclusion criterion. Eight patients responded with strong delayed type hypersensitivity reactions. In 2 of the patients an in vitro response to the vaccine could also be detected. The specificity of the reaction could be confirmed by cloning of peptide-specific CD4 positive T cells from peripheral blood of the patients. Intradermal injection of ras peptides using GM-CSF as adjuvant is simple to perform and seems to be efficient in inducing cellular immune responses. Since a majority of the patients showed positive skin reactions and 2 of the patients analysed showed a T-helper response to this melanoma specific antigen, these promiscuous HLA class II binding mutant ras peptides may be candidates for inclusion into vaccine cocktails containing various established CTL epitopes. PMID- 11380612 TI - Epidermal CD8+ T cells in chronic plaque psoriasis are Tc1 cells producing heterogeneous levels of interferon-gamma. AB - The majority of epidermal CD8+ T cells in chronic plaque psoriasis are activated Tc1 cells producing interferon-gamma and no interleukin-4, a small proportion of which express NK-T receptors. To quantitate their level of cytokine production and characterize them further, CD8+ T cells were isolated from epidermal cell suspensions of lesional biopsies from 24 patients with chronic plaque psoriasis. T-cell lines (TCL) were established by culture of CD8+ T cells with feeders and IL-2 for 11 days and expansion with PHA. Ten TCL were stained for surface markers; 6 were cloned with PHA by limiting dilution. Interferon-gamma, interleukin-4 and interleukin-10 production was measured by ELISA after PMA/anti CD3 activation of 15 TCL and 39 CD8+ T-cell clones. The 10 TCL stained were CD8alphabeta+ (93.3%), T-cell receptor-alphabeta+ (99.5%), costimulatory molecule CD28+ (90.1%), with a small CD8alphaalpha+ population (2.3%). No NK-T-cell receptor CD158a or CD158b expression was detected, whilst CD94 was expressed on 6.2% of cells in 6/9 TCL. All the TCL and 37/39 CD8+ T-cell clones produced interferon-gamma but no or minimal interleukin-4 or interleukin-10. The TCL produced a wide range of interferon-gamma levels (138 to 15,020 pg/ml). Clones from 3 patients showed low levels (60 to 1,410 pg/ml), from 2 patients high levels (6,105 to 43,040 pg/ml) and from 1 patient a wide range (405 to 36,010 pg/ml) of interferon-gamma production. Thus epidermal CD8+ Tc1 cells in chronic plaque psoriasis produce highly heterogeneous levels of interferon-gamma, which may reflect clinical diversity. PMID- 11380613 TI - Metalloelastase (MMP-12) and 92-kDa gelatinase (MMP-9) as well as their inhibitors, TIMP-1 and -3, are expressed in psoriatic lesions. AB - In skin biology, matrix metalloproteinases (MMPs) have been implicated in inflammatory matrix remodeling, neovascularization, wound healing and malignant transformation. Psoriasis is histologically characterized by keratinocyte hyperproliferation, infiltration of inflammatory cells, neoangiogenesis and production of cytokines, such as TNF-alpha, IL-1beta, TGF-alpha, and IFN-gamma, also capable of regulating MMP transcription. To investigate the role of stromelysins-1 and -2, matrilysin, metalloelastase, collagenases-1 and -3 and 92 kDa gelatinase as well as their inhibitors, TIMPs-1 and -3, in psoriasis, we performed in situ hybridization using 35S-labeled cRNA probes on 29 psoriatic lesions and 9 samples of normal looking skin from psoriatic patients. Metalloelastase mRNA was detected in 21/27 samples in macrophages that had migrated into the epidermis or in the inflammatory infiltrates of the superficial dermis. A quantity of 92-kDa gelatinase was found in macrophages and neutrophils (25/27). Stromelysin-1 mRNA was detected in basal keratinocytes in 4/21 lesions. Intracellular laminin-5 immunosignal in basal keratinocytes of the same samples, suggested that stromelysin-1 might participate in remodeling of the basement membrane zone. No signal for stromelysin-2 or collagenase-3 was found and only sweat glands were positive for matrilysin. TIMP-1 was more abundantly expressed than TIMP-3 in the inflammatory infiltrates and endothelial cells of dermal papillae (22/29). TIMP-3 was expressed perivascularly in 9/16 samples. Our results suggest that overexpression of the investigated MMPs by keratinocytes is not associated with psoriasis. However, macrophages express MMPs in psoriatic skin. Also TIMPs, particularly TIMP-1, were abundantly expressed, suggesting that mere MMP overexpression is unlikely to contribute to psoriatic tissue changes. PMID- 11380614 TI - Inhibition of keratinocyte growth in cell culture and whole skin culture by mast cell mediators. AB - Mast cells are suggested to participate in regenerative processes, but their influence on epithelialization and wound healing has not been well studied. Since mast cells can be found in contact with epidermis in chronic inflammatory skin diseases and venous ulcers, the effect of mast cells on keratinocyte growth was studied. Keratinocytes were cultured in serum-free conditions with (complete medium) or without (basal medium) epidermal growth factor (EGF) and bovine pituitary extract (BPE) to reach subconfluence in a 24-well plate, and the cells were treated with different mast cell mediators histamine, heparin and tryptase, or lysate from HMC-1 cells, a human leukemic mast cell line. Whole skin cultures were used as a model for in vitro wounds to study the effect of mast cells on epithelial outgrowth from skin specimens. Histamine inhibited 3H-thymidine incorporation of keratinocytes dose-dependently by 29% at 1 mM, and 89% at 5 mM histamine. In whole skin culture, histamine inhibited epithelial outgrowth dose dependently by 64% already at 0.1 mM histamine and maximally (91%) at 1 mM histamine. Heparin inhibited 3H-thymidine incorporation dose-dependently by up to 33% at 2 microg/ml in the absence, but not in the presence, of EGF/BPE. In contrast, in whole skin culture, heparin first inhibited the epithelial outgrowth by up to 27% at 2 microg/ml, but then reversed the inhibition to 30% stimulation at 200 microg/ml. Skin tryptase (0.0285 to 2.85 microg/ml) with or without heparin (0.5 to 20 microg/ml) did not affect thymidine incorporation in keratinocytes. Lysate from HMC-1 cells, but not that from control, neuroblastoma cells, inhibited 3H-thymidine incorporation in keratinocytes dose-dependently, and maximal (47%) inhibition was reached with 16,700 lysed HMC-1 cells/ml. In whole skin culture, HMC-1 lysate inhibited the epithelial outgrowth by up to 36% at 67,000 lysed cells/ml. The results show that mast cells and their mediators are inhibitory to keratinocyte 3H-thymidine incorporation and epithelial outgrowth in vitro, although, the inhibitory effect of histamine was seen at high concentrations suggesting a requirement for close morphologic vicinity of mast cells to keratinocytes. Thus, mast cells are assumed to control epidermal regeneration and to impair epithelialization of chronic ulcers. PMID- 11380615 TI - Proprotein convertase expression and localization in epidermis: evidence for multiple roles and substrates. AB - Specific proteolysis plays an important role in the terminal differentiation of keratinocytes in the epidermis and several types of proteases have been implicated in this process. The proprotein convertases (PCs) are a family of Ca2+ dependent serine proteases involved in processing and activation of several types of substrates. In this study we examined the expression and some potential substrates of PCs in epidermis. Four PCs are expressed in epidermis: furin, PACE4, PC5/6 and PC7/8. Furin is detected in two forms, either with or without the transmembrane domain, suggesting occurrence of post-translational cleavage to produce a soluble enzyme. In addition the furin active site has differential accessibility in the granular layer of the epidermis relative to the basal layer, whereas antibodies to the transmembrane domain stain both layers. These findings suggest that furin has access to different types of substrates in granular cells as opposed to basal cells. PC7/8, in contrast, is detected throughout the epidermis with antibodies to both the transmembrane and active site and no soluble form observed. A peptide PC inhibitor (dec-RVKR-CMK) inhibits cleavage of Notch-1, a receptor important in cell fate determination that is found throughout the epidermis. Profilaggrin, found in the granular layer, is specifically cleaved by furin and PACE4 in vitro at a site between the amino terminus and the first filaggrin repeat. This work suggests that the PCs play multiple roles during epidermal differentiation. PMID- 11380616 TI - UV-A-induced AP-1 activation requires the Raf/ERK pathway in human NCTC 2544 keratinocytes. AB - UV-A irradiation causes a dose-dependent activation of ERK in human NCTC 2544 keratinocytes. The specific inhibition of either ERK activity or Raf kinase activity impedes the activation of AP-1 DNA binding induced by UV-A. In addition, UV-A raises AP-1 promoter transcriptional activity, which is downregulated in NCTC 2544 cells expressing an inactive mutant of Raf-1. We found that singlet oxygen might be one of the mediators in both UV-A-induced AP-1 DNA binding and transcriptional activity. These results strongly suggest that UV-A-induced AP-1 activity requires the Raf-ERK pathway and imply a singlet oxygen effector. PMID- 11380617 TI - Conformational switching of Escherichia coli RNA polymerase-promoter binary complex is facilitated by elongation factor GreA and GreB. AB - BACKGROUND: The initiation arrest at a modified lambdaPR promoter is caused by irreversible divergence of the reaction pathway into productive and arrested branches. Escherichia coli GreA and GreB induce cleavage of the nascent transcript and relieve arrest in elongation. They also reduce abortive synthesis at several promoters and relieve initiation arrest. Their mechanism of action during initiation, and its relationship to the branched initiation pathway are unknown. RESULTS: The Gre factors mitigated initiation arrest only when they were added to the binary complex of the holoenzyme bound to the lambdaPR promoter, prior to RNA synthesis. They exerted little effect when they were added to ternary initiation complexes. They accelerated the exchange of the binary complex with its free components by 6-9-fold. When they are present, a high concentration of the initiating nucleotide increased yield of the full-length transcript, whereas a low concentration did not. CONCLUSIONS: All the results presented above can be explained by a model where the productive and arrested pathways diverge at the binary complex stage. The Gre factors relieve the initiation arrest by introducing reversibility between subspecies of the binary complex that are precursors of the two pathways. RNA cleavage is unlikely to cause relief of initiation arrest. PMID- 11380618 TI - Involvement of RNase G in in vivo mRNA metabolism in Escherichia coli. AB - BACKGROUND: Escherichia coli rng gene (previously called cafA) encodes a novel RNase, named RNase G, which is involved in the 5' end-processing of 16S rRNA. In rng mutant cells, a precursor form of 16S rRNA, 16.3S rRNA, is accumulated. Here we report a role of RNase G in the in vivo mRNA metabolism. RESULTS: We found that rng:cat mutant strains overproduced a protein of about 100 kDa. N-terminal amino acid sequencing of this protein showed that it was identical to the fermentative alcohol dehydrogenase, the product of the adhE gene located at 28 min on the E. coli genetic map. The level of adhE mRNA was significantly higher in the rng:cat mutant strain than that in its parental strain, while such differences were not seen in other genes we examined. A rifampicin-chase experiment revealed that the half-life of adhE mRNA was 2.5-fold longer in the rng:cat disruptant than in the wild-type. CONCLUSION: These results indicate that, in addition to rRNA processing, RNase G is involved in in vivo mRNA degradation in E. coli. PMID- 11380619 TI - Self-splicing of the Tetrahymena group I ribozyme without conserved base-triples. AB - BACKGROUND: Group I introns share a conserved core region consisting of two domains, P8-P3-P7 and P4-P6, joined by four base-triples. We showed previously that the T4 td intron can perform phosphoester transfer reactions at two splice sites in the absence of both P4-P6 and the conserved base-triples, whereas it is barely able to perform the intact splicing reaction due to the difficulty of conducting the sequential reactions. RESULTS: Based on previous findings, we constructed a bimolecular ribozyme lacking a large portion of P4-P6 and the base triples from the Tetrahymena intron, on the assumption that the long-range interactions of the peripheral regions in the two RNAs can compensate for the deteriorated core. The bimolecular ribozyme performed the intact splicing reaction. CONCLUSION: The present analysis indicates that the base-triples are nonessential, but that L4 and the distal part of P4 in P4-P6 are important for conducting the splicing reaction. The reconstituted self-splicing ribozyme provides an amenable system for analysing the role(s) of elements in the core region in the self-splicing reaction mechanism. PMID- 11380620 TI - Werner helicase relocates into nuclear foci in response to DNA damaging agents and co-localizes with RPA and Rad51. AB - BACKGROUND: Werner syndrome (WS) is an autosomal recessive disorder with many features of premature ageing. Cells derived from WS patients show genomic instability, aberrations in the S-phase and sensitivity to genotoxic agents. The gene responsible for WS (WRN) encodes a DNA helicase belonging to the RecQ helicase family. Although biochemical studies showed that the gene product of WRN (WRNp) interacts with proteins that participate in DNA metabolism, its precise biological function remains unclear. RESULTS: Using immunocytochemistry, we found that WRNp forms distinct nuclear foci in response to DNA damaging agents, including camptothecin (CPT), etoposide, 4-nitroquinolin-N-oxide and bleomycin. The presence of aphidicolin inhibited CPT-induced WRNp foci strongly but not bleomycin-induced foci. These WRNp foci overlapped with the foci of replication protein A (RPA) almost entirely and with the foci of Rad51 partially, implicating cooperative functions of these proteins in response to DNA damage. We also found that WRNp foci partially co-localize with sites of 5-bromo-2'-deoxy-uridine incorporation. CONCLUSIONS: These findings suggest that WRNp form nuclear foci in response to aberrant DNA structures, including DNA double-strand breaks and stalled replication forks. We propose that WRNp takes part in the homologous recombinational repair and in the processing of stalled replication forks. PMID- 11380621 TI - Crk family adaptor proteins trans-activate c-Abl kinase. AB - BACKGROUND: c-Abl kinase is activated in response to a variety of biological stimuli. Crk family adaptor proteins can interact physically with c-Abl and be involved in the activation of c-Abl kinase. RESULTS: We report that the Crk family of adaptor proteins act as trans-acting activators of c-Abl kinase. The interaction of the amino-terminal Src-homology (SH) 3 domain of c-Crk and the proline-rich motifs of c-Abl is an essential step for the phosphorylation of c Crk by c-Abl, as well as the activation of c-Abl by c-Crk. The activation of c Abl by c-Crk is negatively regulated by phosphorylation of the tyrosine 221 of c Crk. Our data suggest that, in the absence of phosphorylation of the tyrosine Y221, the SH2 domain of c-Crk becomes free to bind to target molecules while the carboxyl-terminal SH3 domain of c-Crk binds to the proline-rich region of c-Abl, inducing the activation of c-Abl by c-Crk. CONCLUSION: This study suggests that the Crk family functions as trans-acting activators of c-Abl kinase. The phosphorylation of c-Crk may regulate c-Abl kinase. PMID- 11380622 TI - Identification of the cis-acting region in the NF2 gene promoter as a potential target for mutation and methylation-dependent silencing in schwannoma. AB - BACKGROUND: Although mutational inactivation and allelic loss in the NF2 gene appear to be causal events in the majority of vestibular schwannomas, involvement of another potentially important mechanism, transcriptional inactivation, has not been investigated. RESULTS: We cloned and functionally characterized the 5' flanking region of the human NF2 gene and identified the molecular mechanisms that regulate NF2 expression. Luciferase assay and site-directed mutagenesis demonstrated that a 70-base pair (bp) region (-591 to -522 bp from the translation start site) was essential for the basic expression of the NF2 gene. A gel mobility shift assay indicated recognition by nuclear protein of the unusually long ( approximately 66 bp) sequences in this region. Recognition was inhibited by either mutation of the binding core sequence or by methylation of three CpG sites. Point mutations at these CpG sites significantly decreased promoter activity, suggesting the importance of these sites. In 14 of 23 vestibular schwannomas, these three CpG sites were methylated in a site-specific manner and the methylation status was consistent with the expression of NF2 mRNA. CONCLUSIONS: Suppressed expression by aberrant methylation or mutation of the promoter elements could be an alternative mechanism for inactivation of the NF2 gene. PMID- 11380623 TI - Fission yeast homologues of the B' subunit of protein phosphatase 2A: multiple roles in mitotic cell division and functional interaction with calcineurin. AB - BACKGROUND: Protein phosphatase 2A (PP2A) is a serine/threonine phosphatase distributed in eukaryotes from yeast to human, and plays pivotal roles in diverse cellular functions such as metabolism, cell cycle progression, gene expression and development. PP2A holoenzyme is a heterodimer of a catalytic subunit C and a regulatory subunit A, or a heterotrimer of C, A and a variable regulatory subunit consisting of three families; B, B', and PR72. Specific functions for each variable subunit are not well understood. RESULTS: Two fission yeast genes pbp1+ and pbp2+ homologous to the regulatory subunit B' were isolated. Physical in vivo interaction of the gene products with the catalytic subunit was demonstrated. A double disruption haploid mutant (Deltapbp1Deltapbp2) showed growth defect, cell shape and size abnormality, multiseptation and anucleated cell formation due to abnormality in septum positioning. These phenotypes were suppressed by human B' cDNA, indicating the striking conservation of the B' function from yeast to human. Over-expression of fission yeast B' led to growth defects, a loss of cell shape polarity, septal abnormality and anucleated cell formation. Deltapbp1Deltapbp2 and pbp1 null haploids were hypersensitive to calcineurin inhibitors, cyclosporin A and FK506, with which the mutants underwent arrest at post-anaphase and cell lysis. Double disruption of calcineurin and pbp1+, but not pbp2+, genes led to synthetic lethality. CONCLUSION: The fission yeast B' subunit of PP2A plays critical roles in cell shape control and septum formation, and shares essential functions with calcineurin for viability, possibly through their roles in cytokinesis and cell wall integrity. PMID- 11380625 TI - The central MHC gene, BAT1, may encode a protein that down-regulates cytokine production. AB - BACKGROUND: BAT1 belongs to the DEAD-box family of RNA-binding proteins and is encoded in the central MHC. To determine whether it affects immune responses and hence diseases influenced by MHC haplotypes, U937, THP1 and Jurkat cells were stably transfected with anti-sense DNA corresponding to exons 2-5 of BAT1 using a retroviral vector. RESULTS: Anti-sense transfectants carried anti-sense DNA and expressed anti-sense mRNA. After mitogenic stimulation, they produced higher levels of TNFalpha, IL-1 and IL-6 than equivalent cells carrying the vector alone, suggesting that BAT1 may down-regulate acute phase cytokine production. Polyclonal antibodies raised against a peptide in exon 8 of BAT1 recognized approximately 50 kDa and approximately 38 kDa proteins in all cell lines tested, including the anti-sense transfectants. Expression was localized to the nucleolus in dividing fibroblasts. However the immunochemistry may be confounded by a recently described gene, DDXL, on chromosome 19, which shares a 89% amino acid identity with BAT1. RT-PCR analyses established that BAT1 and DDXL mRNA are expressed in resting U937, THP1 and Jurkat cells. BAT1 and DDXL are divergent in the exons selected for the anti-sense study. CONCLUSIONS: BAT1 is a negative regulator of inflammation. Future studies should address how its functions relate to those of DDXL. PMID- 11380624 TI - Requirement of Syk-phospholipase C-gamma2 pathway for phorbol ester-induced phospholipase D activation in DT40 cells. AB - BACKGROUND: Treatment of many cell types with phorbol esters stimulates phospholipase D (PLD) activity implying regulation of the enzyme by protein kinase C. Studies of the effects of several protein-tyrosine kinase (PTK) inhibitors have suggested that PTK(s) play some roles in the phorbol ester induced PLD activation, but it remains unclear how and which PTK(s) is involved in this pathway. In this study, we investigated the roles of Syk and other PTKs for the phorbol esters, 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced PLD activation in K562 and DT40 cells. RESULTS: TPA-induced PLD activation was remarkably reduced in both Syk dominant negative mutant K562 cells and Syk deficient DT40 B cells. Mutational analysis further indicated that two major autophosphorylation sites (Tyr-518 and Tyr-519) of Syk are critical for PLD activation. Similarly, TPA-induced PLD activation was reduced in Btk deficient cells, but unaffected in Lyn deficient cells. Finally, in cells deficient in the PLC-gamma2, one of the phosphorylated substrates regulated by Syk and Btk, TPA induced PLD activation, as well as phosphatidylinositol 4,5-bisphosphate (PIP2) hydrolysis was remarkably reduced. CONCLUSIONS: We demonstrated that the Syk, Btk and PLC-gamma2 pathways are required for TPA-induced PLD activation in DT40 cells. PMID- 11380626 TI - Management of von Willebrand disease: a survey on current clinical practice from the haemophilia centres of North America. AB - The optimal treatment of patients with von Willebrand's disease (vWD) remains to be defined. Moreover, it has not been firmly established which, if any, commonly measured parameters of von Willebrand factor (vWF) protein in the plasma are useful in guiding treatment. To better understand what guidelines physicians follow in the management of vWD, we surveyed 194 North American physicians who are members of the Hemophilia Research Society. Ninety-nine per cent of responding physicians depend on factor VIII (FVIII):C, vWF:RCo activity and vWF:AG to diagnose vWD, while only 49% use the bleeding time. The minimal goals of treatment for patients undergoing major surgery/trauma or central nervous system haemorrhage were FVIII:C and vWF:RCo activity greater than 80% while levels of more than 50% for minor surgery and dental extractions were considered adequate. Treatment of vWD was based on the type of vWD with type 1 patients being treated most often with desmopressin acetate (DDAVP) alone, types 2A and 2B patients with a combination of DDAVP and a vWF-containing FVIII product, type 3 patients with vWF-containing concentrate. Viral infections, including human immunodeficiency virus, hepatitis A, B and C viruses, and parvovirus have been seen in vWD and the efficacy of viral attenuation processes is a major criterion for the selection of treatment by physicians. Based on this survey, prospective studies need to be designed to address the clinical efficacy, safety and predictive value of laboratory monitoring of patients with vWD. PMID- 11380628 TI - Vitronectin in clotting factor IX concentrates. AB - Highly purified, plasma-derived factor IX (FIX) concentrates are produced in large part by a combination of anion exchange and heparin affinity chromatography. However, the concentrates still contain some accompanying proteins. The main impurity has turned out to be the adhesive glycoprotein, vitronectin. It occurs in concentrates exclusively in its multimeric form, in contrast to the situation in plasma. The multimeric vitronectin can be removed either by nanofiltration with a crossflow system or by size-exclusion chromatography. When these FIX concentrates are used as therapeutic agents, the fact has to be taken into account that considerable amounts of multimeric vitronectin are given to the patient. The physiological consequences of the dosage of this protein have not yet been investigated. Although no thrombogenicity has been reported in connection with the above-mentioned FIX concentrates, we recommend that the impurity should be removed from the preparation with the methods described here. PMID- 11380627 TI - Safety and efficacy of a new recombinant FVIII formulated with sucrose (rFVIII FS) in patients with haemophilia A: a long-term, multicentre clinical study in Japan. AB - The recombinant full-length FVIII product Kogenate has been reformulated using sucrose (rFVIII-FS) instead of human serum albumin as a stabiliser in purification and formulation. The in vivo recovery, haemostatic efficacy, and safety of rFVIII-FS were investigated in 20 previously treated patients with severe or moderate haemophilia A for > or = 24 weeks. In vivo recoveries of 73.5 +/- 16.3%, 78.4 +/- 16.1%, and 82.8 +/- 23.9% after the initial infusion of 50 IU kg(-1) rFVIII-FS and at weeks 12 and 24, respectively, showed no significant changes over time. A total of 1115 infusions (mean dose 24.1 +/- 8.4 IU kg(-1)) were included in the analysis of haemostatic efficacy. One (80.5%) or two (8.2%) infusions achieved adequate haemostasis in 88.7% of all bleeding episodes, and haemostatic efficacy was judged 'excellent' or 'good' in 749 of 764 episodes (98.0%). The haemostatic efficacy was judged as 'excellent' or 'good' in 924 of 1115 (82.9%) infusions. Twenty-one adverse events were observed in 12 patients in the total 1541 infusions included in the safety analysis. Causality with respect to rFVIII-FS could not be ruled out in three events in one HIV-negative patient: elevated CD4(%), decreased CD8(%), and elevated CD4/CD8 ratio. No FVIII inhibitor development was observed in any patient. ELISA assay testing for antibodies to rFVIII, baby hamster kidney cell (BHK) protein, and murine IgG were all negative. These results show that rFVIII-FS is a safe and effective for long-term treatment of patients with haemophilia A. PMID- 11380629 TI - High-dose DDAVP intranasal spray (Stimate) for the prevention and treatment of bleeding in patients with mild haemophilia A, mild or moderate type 1 von Willebrand disease and symptomatic carriers of haemophilia A. AB - An open-label multicentre trial was conducted to evaluate high-dose DDAVP (desmopressin acetate) intranasal spray (Stimate; 1.5 mg mL(-1)), for the control of bleeding in 333 patients with mild haemophilia A, mild or moderate type 1 von Willebrand disease, or symptomatic carriers of haemophilia A. Overall, 278 patients received 2170 doses of high-dose DDAVP intranasal spray (1.5 mg mL(-1)). Using study-defined guidelines, patients evaluated the efficacy of high-dose DDAVP intranasal spray (1.5 mg mL(-1)) as 'excellent' or 'good' in 743 (95%) of 784 bleeding episodes. It demonstrated 'excellent' results in 384 (93%) of 413 administrations for prophylaxis and in eight of eight uses prior to acute surgical or dental procedures. When used for the treatment of menorrhagia, the efficacy of high-dose DDAVP intranasal spray (1.5 mg mL(-1)) was rated as 'excellent' after 655 (92%) of 721 daily uses. Of 2170 doses of high-dose DDAVP intranasal spray (1.5 mg mL(-1)), 172 (8%) were associated with adverse events. A total of 272 adverse events were reported among 80 patients. Of these, 239 (88%) were mild or moderate in intensity and only one patient was removed from the study due to an adverse event. These results demonstrate the safety and efficacy of high-dose DDAVP intranasal spray (1.5 mg mL(-1)) for control of bleeding episodes in patients with mildly decreased levels of factor VIII, von Willebrand factor, or both. PMID- 11380630 TI - The Malmo International Brother Study (MIBS): further support for genetic predisposition to inhibitor development in hemophilia patients. AB - The issue of factors predisposing for inhibitor development in haemophilia patients is still largely unresolved. In an attempt to address this problem, we initiated a registry in 1996 of siblings with haemophilia and with or without a history of inhibitors. Four hundred and sixty families have accrued, of whom 388 suffer from haemophilia A and 72 haemophilia B. Twenty-five of the brother pairs are twins. The inhibitor incidence in all families with severe haemophilia A was 31.7%. The corresponding figure in the caucasian patients was 27.4%, whereas a higher incidence of inhibitors was reported in the black subjects (55.6%). Twins were reported in six of the 100 inhibitor families, for whom monozygocity was confirmed in three cases. In 32 families (32%), at least two brothers had a history of inhibitors. In 22 (69%) of these families, the inhibitor was also of the same type, i.e. either high- or low-responding. The overall concordance within the severe haemophilia A families was found to be 78.3% (195/249) compared to an expected figure of 68.0% and 58.0% using an inhibitor incidence of 20 and 30%, respectively (P < 0.0001). The corresponding figure for the twins was 88.2% (15/17). Moreover, the risk for inhibitor development in families with a previous inhibitor history was found to be 48% (95% confidence interval [CI] 35-62%), whereas the risk in families with no previous known inhibitor was only 15% (95% CI 11-21%) corresponding to a relative risk of 3.2 (95% CI 2.1-4.9). Immune tolerance induction was reported in 24 families, of whom 13 siblings were successfully treated. Our data clearly support the concept that a genetic predisposition for inhibitor development exists. However, the markers of this predisposition remain to be elucidated and we believe that the MIBS registry will be useful for this purpose. PMID- 11380631 TI - Development of inhibitors in patients with haemophilia from India. AB - Four hundred and seven patients (352 haemophilia A and 55 haemophilia B) were investigated for the presence of factor VIII and IX inhibitors. Twenty-four out of 292 severe and two out of 36 moderate haemophilia A patients showed the presence of inhibitors. The mean age at development of inhibitors was 17.7 years (range 6-52 years). In 12 patients the inhibitors were detected due to suboptimal response to factor replacement therapy (symptomatic) and in the remaining 14 patients the inhibitors were detected during the routine screening of the patients' samples for inhibitors. They had, however, responded well to the usual doses of factor concentrates and there was no suspicion in these patients that they had developed an inhibitor (asymptomatic). There were two families in which the inhibitors were detected in more than one family member. The level of inhibitors in symptomatic patients ranged from 2.2 Bethesda units (BU) mL(-1) to 460.6 BU mL(-1), and in asymptomatic patients it ranged from 0.8 BU mL(-1) to 3.2 BU mL(-1). The inhibitors persisted in all patients except one, who developed an inhibitor postoperatively for a brief period of 3 months. All these patients were followed up from first factor exposure and were tested for inhibitors at least twice a year. The mean number of exposure days before they developed inhibitors was 47.5 exposure days (range 17-98 exposure days). No inhibitors appeared after more than 100 exposure days in any of the patients. When 50 consecutive patients were investigated for intron 22 inversions of the factor VIII gene, 17 patients were found to be positive for inversions (10 proximal inversion; seven distal inversion) out of whom four patients developed inhibitors, three patients belonging to the same family. Out of 35 haemophilia B patients, only one patient developed an inhibitor. The overall prevalence of inhibitors was thus 8.2%, which is similar to the reports from western countries, prior to the introduction of highly purified factor concentrate therapy. PMID- 11380632 TI - Cost-utility analysis of recombinant factor VIIa (NovoSeven) in six children with long-standing inhibitors to factor VIII or IX. AB - The high cost of treating patients with inhibitors in an environment of restricted budgets warrants consideration of cost-effectiveness. We determined the clinical response, effect on quality of life and the cost-effectiveness of treatment with rFVIIa in six boys with long-standing inhibitors to factors VIII or IX, compared with other treatment regimes previously used in these patients. The study used a longitudinal before-and-after design and was conducted in three phases. Phase 1 was 6 months preceding the introduction of rFVIIa, during which patients received on-demand 'usual care' with other treatment regimes; phase 2 was 6 months treatment on rFVIIa assessed retrospectively; and phase 3 was 6 months on rfVIIa treatment assessed prospectively. Treatment with rFVIIa was reserved for intrarticular, compartment, psoas, mucosal and suspected intracranial bleeding. Treatment outcomes were obtained by interview using structured questionnaires, the quality-of-life instruments CHQ CF-80 and CHQ PF 50, patient self-reporting diary, interrogation of hospital records, and the EuroQoL EQ-5D for utility valuations. Our results confirm that rFVIIa is clinically effective and resulted in 63-92% reductions in the number of re treatments, duration of painful episodes, delay to initiation of treatment, days requiring wheelchair or crutches, emergency room visits and lost carer time compared with the patients' other therapies. Quality-of-life improvements were observed in several important areas as perceived by both patients and their families, at an incremental cost per QALY of A$51 533. PMID- 11380633 TI - Correlation between clinical, radiological and ultrasonographical image of knee joints in children with haemophilia. AB - The aims of the study were to evaluate the clinical, radiological and ultrasonographical images of knee joints in children with severe haemophilia and von Willebrand's disease, to determine the correlation between these images and to assess the usefulness of ultrasonography (USG) in evaluating the intensity of haemophilic arthropathy. Thirty-nine boys were included in the study, all of them with a past history of knee bleeds. The average age of the children was 10.02 +/- 3.01 years. In patients with slight (1-3 points) and moderate (4-7 points) radiological changes in knee joint bones, an increase in synovial fluid, considerable hypertrophy and inflammation of the synovium were observed in USG. In haemophilic patients with severe (8-13 points) bone changes, the amount of fluid was usually normal and there was slight inflammation but considerable hypertrophy of the synovium. Radiological evaluation of haemophilic arthropathy was made according to the Pettersson classification. A good correlation between the degree of cartilage damage in USG and the progression of bone changes in radiographs was found. Cartilage and bone damage progressed with the increase in the number of intra-articular haemorrhages into the knee joint. In our opinion USG is useful in evaluating the fluid, synovium and cartilage of joints in haemophiliacs. PMID- 11380634 TI - Motor performance and disability in Dutch children with haemophilia: a comparison with their healthy peers. AB - We investigated whether haemophilic children who are on prophylactic therapy differ from their healthy peers in terms of motor performance and disability. Thirty-nine children, aged 4-12 years, with moderate (eight) and severe (31) haemophilia were included. Patients with severe haemophilia received primary prophylactic therapy that was individually tailored. The number of target joints, amount of swelling, range of motion, muscular strength and pain were measured, as well as motor skills and disability. The scores were compared to the normal population. No patients had target joints. Normal range of motion in all joints was seen in 97% (38/39) of the patients. Strength of elbow, knee, and ankle muscles were within the normal ranges. Ninety-five percent (37/39) of the patients had normal motor performance. Although 90% of our patients (35/39) had no disabilities in activities of daily living (ADL), 79% (31/39) of them reported that the disease impacted on their lives. Seventy-two percent (28/39) of the patients had pain, and in 21% (6/28) of them this was mainly caused by injections. Restrictions in sports or gymnastics were seen in 56% (22/39) of the patients. Those who indicated that they experienced pain and those who indicated restrictions in sports had a higher chance of experiencing disease impact compared to those who did not have these limitations. There were no significant differences between patients with moderate and severe haemophilia. In general, Dutch children with moderate or severe haemophilia are comparable with their healthy peers with regard to motor performance and ADL. However, a majority of the patients perceive an impact of their disease associated with pain and restrictions in sports. PMID- 11380635 TI - Factor VIII gene inversions and polymorphisms in Brazilian patients with haemophilia A: carrier detection and prenatal diagnosis. AB - In families afflicted with haemophilia A, genetic counselling is often requested. Inversion mutations and polymorphic sites of the FVIII gene have been examined in a Brazilian population, with the aim of developing a strategy that would be accurate and informative for carrier analysis and prenatal diagnosis in Brazil. Patients with haemophilia A and families were studied. Inversion mutations in the FVIII gene were detected in 39.4% of severely affected patients, 85% of them being of distal type. No inversions were observed in patients with mild or moderate forms of the disease. Two bi-allelic polymorphisms were studied. Intron 18 SSCP and intron 19 RFLP analyses indicated the presence of a restriction site in 39.5% and 42.9% of haemophilics, respectively. Two multiallelic microsatellite polymorphisms in introns 13 and 22 were also studied; eight different alleles were detected in each system with a heterozygosity rate of 83.08% and 78.77%, respectively. When all four intragenic loci were examined in linkage analysis, the cumulative reliability was 100%. In conclusion, inversion mutation analysis should be the first-line test for Brazilian patients with severe haemophilia A. In cases of severe haemophilia A where no inversion could be detected or in mild or moderate haemophilia A, the use of all four polymorphisms is very informative for linkage analysis and should be used for carrier detection and genetic counselling in the Brazilian population. PMID- 11380636 TI - A haemophilia A and B molecular genetic diagnostic programme in Hungary: a highly informative and cost-effective strategy. AB - Our aim was to set up a protocol in order to provide carrier and prenatal diagnosis to Hungarian haemophilia A (HA) and B (HB) patients and their relatives. For HA, a combination of direct mutation detection and some indirect marker analyses were used: the detection of the inversion mutation and analysis of three polymorphisms, BclI, IVS13 (CA)n and P39(CA)n. In severe cases, direct mutation detection was performed first. In inversion-negative severe cases and in moderate and mild cases, indirect methods were used. For carrier and prenatal diagnosis in HB, four polymorphisms, DdeI, TaqI, XmnI, and HhaI were examined. Our DNA bank contains samples from 50 HA families (34 severe, 15 moderate and one mild) and seven HB families from different parts of the country. In 100% of the HA cases either the gene inversion and/or at least one of the polymorphisms was found to be informative for carrier or prenatal diagnosis. In the HB cases, an informative marker was found in 95% of the cases (19 of 20). We conclude that these strategies are sufficient to make genetic diagnosis available to almost all HA and HB families in the region. This approach is highly informative and cost effective, so it can be very useful in countries where direct sequencing of genes for factor VIII and IX is not available for routine diagnosis. PMID- 11380637 TI - Knee joint arthroplasty in a patient with haemophilia A and high inhibitor titre using recombinant factor VIIa (NovoSeven): a new case report and review of the literature. AB - Elective orthopaedic surgery is regularly withheld from patients with haemophilia and high inhibitor titre despite the presence of severe arthropathy and urgent medical need. A knee joint arthroplasty was performed in a patient with severe haemophilia A and a high inhibitor titre using recombinant factor VIIa (rFVIIa) as the sole coagulation factor. There was no abnormal bleeding during surgery although an increased blood loss through surgical drains did occur during the first 6 h postoperatively. Rehabilitation was started on day 1 and continued for 3 months. Walking commenced on day 4. After 1 year of follow-up, the clinical outcome of surgery was considered excellent with no pain, knee mobility at 0-5-90 degrees, and an International Knee Society score of 95/100. No rFVIIa-associated side-effects or thrombotic complications were reported. In conclusion, knee joint arthroplasty is now an option for haemophilia patients with a high inhibitor titre. An international review of all available data on elective orthopaedic surgery in inhibitor patients is required so that the optimal treatment regime can be defined and the short- and long-term risk-benefit ratio of surgery compared to that of noninhibitor patients. PMID- 11380638 TI - Postpartum acquired haemophilia: clinical recognition and management. AB - Postpartum acquired haemophilia is a rare but serious complication of an otherwise normal pregnancy. Patients usually present with postpartum haemorrhage (PPH) or uncontrolled bleeding following surgical interventions, which fail to respond to conservative treatment. A high index of clinical suspicion along with early laboratory diagnosis and prompt institution of appropriate therapy is essential for the management of acute bleeding episodes. Our patient, a 32-year old female, presented with severe PPH and shock. She had undergone dilation and curettage three times, with subsequent total abdominal hysterectomy and internal iliac artery ligation, before she was diagnosed with acquired haemophilia (factor VIII autoantibodies) and an inhibitor level of 8 Bethesda units (BU). The patient underwent an abdominal laparotomy for removal of the abdominal packing used in the previous operation, and blood and blood clots, and was given FEIBA(R) therapy. The patient responded to these measure and the factor VIII inhibitor level decreased to 2 BU at the time of discharge 10 weeks later. PMID- 11380639 TI - Blunt abdominal trauma with delayed rupture of splenic haematoma in a haemophiliac patient. AB - A 13-year-old haemophilia A patient presented with pain in the abdomen, 4 days after a blunt abdominal trauma. The computed tomography scan of the abdomen showed a large splenic haematoma. The patient was initially managed with factor VIII replacement therapy, but 4 weeks later he had a delayed rupture of the splenic haematoma with haemoperitoneum and shock. An elective splenectomy under factor VIII therapy was successfully performed. PMID- 11380640 TI - Mutation causing exon 15 skipping and partial exon 16 deletion in factor VIII transcript, and a method for direct mutation detection. AB - A splicing defect with 201 nucleotide deletion in the factor VIII transcript due to IVS15 + 1G > T mutation inactivating this donor splice site and activating a cryptic acceptor splice site in exon 16 was identified in a severe haemophilia A patient. Allele specific amplification (ASA) method was successfully developed for direct detection of this mutation. PMID- 11380641 TI - Evaluating HIV-infected patients with headache: who needs computed tomography? AB - OBJECTIVE: To empirically test a clinical prediction rule for evaluating HIV infected patients complaining of headache and to identify those at low risk for intracranial mass lesion who do not need immediate computed tomography of the head. DESIGN: Two retrospective clinical cohorts of HIV-infected patients clinically evaluated for headache. METHODS: To describe the headache clinical outcomes, medical records were abstracted from all HIV-infected patients evaluated for headache with computed tomography of the head at two urban hospitals. Patients were categorized as low, intermediate, or high risk based on clinical criteria (focal neurological signs, altered mental status, history of seizure) and immune status (CD4 lymphocytes < or =200 microL). Records were abstracted from a second unselected cohort of HIV-infected outpatients with headache who were all treated and followed in primary care (N=101). RESULTS: Of 101 unselected HIV-infected outpatients followed in primary care after headache, 1% (95% confidence interval [CI], 0% to 6%) had a treatable intracranial lesion. Of 364 HIV-infected patients with headache sent for evaluation with computed tomography of the head, the rate of any abnormality was zero in the low-risk group (95% CI, 0% to 10%; n=35); 9% in the intermediate-risk group (95% CI, 2% to 16%; n=242); and 21% in the high-risk group (95% CI, 12% to 29%; n=87). CONCLUSION: Most HIV-infected patients with headache may be treated with analgesics and followed up clinically. Those without focal neurological signs, altered mental status, seizure, or decreased CD4 lymphocytes are unlikely to have intracranial mass lesions. PMID- 11380642 TI - Oral almotriptan in the treatment of migraine: safety and tolerability. AB - OBJECTIVE: To summarize safety and tolerability data on orally administered almotriptan from premarketing clinical trials. BACKGROUND: Almotriptan is a new 5 HT1B/1D receptor agonist similar to sumatriptan in mode of action and therapeutic efficacy. In addition, the safety and tolerability profile of almotriptan has been demonstrated in a number of controlled clinical trials. Sumatriptan is generally safe and well tolerated; however, in controlled clinical trials, it has been associated with chest symptoms (pressure, warmth, and other unpleasant sensations) with an incidence of 3% to 5%. DESIGN: Three phase 1 dose-finding and pharmacokinetic studies in healthy men and women volunteers were reviewed to assess the safety and tolerability of oral almotriptan at single doses ranging from 2 to 200 mg. The objective of one study was to evaluate cardiovascular safety. Two phase 2 trials assessed the safety and tolerability of single doses of 2 to 150 mg in migraine (n=911). Two phase 3 trials assessed the safety and tolerability of a single 12.5-mg oral dose after three attacks (n=910) and repeated doses of 12.5 mg for multiple attacks over the long term (n=747). All studies were conducted in Europe. Data from the United States is currently being analyzed and will be published at a later date. RESULTS: In phase 2 and 3 trials comprising more than 2500 patients with migraine and 15 000 attacks, adverse events were infrequent and mild. The most common events-dizziness, nausea and vomiting, headache, fatigue, paresthesia, and drowsiness-were reported in fewer than 3% of patients. At the recommended therapeutic dose of 12.5 mg, the adverse events profile was not statistically different from placebo. The incidence of chest symptoms was 0.2% in the phase 3 trials. The long-term safety and tolerability profile after treatment of more than 10 000 attacks was similar to that following the single-dose studies. In all clinical trials, almotriptan demonstrated a very favorable adverse event profile, particularly with respect to nonischemic-related chest symptoms. CONCLUSIONS: Almotriptan was safe and well tolerated in nearly all adult patients with migraine, with and without aura, enrolled in these studies. The incidence of chest symptoms in preclinical studies was substantially lower than that reported for sumatriptan in premarketing studies, indicating that almotriptan may be better tolerated than sumatriptan at clinically anticipated doses. However, any potential difference in cardiovascular safety between almotriptan and sumatriptan cannot be determined or inferred from this data. Cardiovascular risk profiles for all drugs within this class (triptans) should be considered similar. Only extensive postmarketing data, not currently available, can potentially change this recommendation. PMID- 11380643 TI - Treatment of migraine in Canada with naratriptan: a cost-effectiveness analysis. AB - OBJECTIVE: To evaluate the cost-effectiveness of naratriptan for the treatment of migraine in Canada. BACKGROUND: The substantial disability brought on by migraine, coupled with the high prevalence of this disorder, leads to substantial costs. Naratriptan is a newly developed triptan shown to be effective in the treatment of migraine. METHODS: Monte Carlo modeling techniques were used to simulate the experience of Canadian migraineurs over the course of 1 year. Data from a multinational study comparing oral naratriptan 2.5 mg to customary therapies were used in the cost-effectiveness analysis. RESULTS: Naratriptan leads to an annual reduction in symptom duration of 225 hours compared to customary therapy not including other triptans. Reductions in lost productivity yield savings of Can $390 (1998 Canadian dollars) relative to customary therapy, which exceed the increase in drug costs resulting in overall savings of Can $109 per year. CONCLUSIONS: The use of naratriptan in the treatment of migraine is an economically attractive option, leading to savings in overall costs. Increases in drug costs seem acceptable in light of reductions in symptom duration. PMID- 11380644 TI - Nefazodone for chronic daily headache prophylaxis: an open-label study. AB - OBJECTIVE: To assess effectiveness, tolerability, and safety of nefazodone as a prophylactic agent for chronic daily headache. BACKGROUND: Nefazodone is a potent, selective 5-HT2 antagonist with a distinct and atypical mechanism of action. The evolution of intermittent migraine to chronic daily headache has been linked to up-regulation of 5-HT2 receptors as well as other factors. Other effective migraine prophylactic medications are also 5-HT2 antagonists. Although research has shown nefazodone to be an effective antidepressant with a good tolerability and safety profile, its potential role in headache prophylaxis has not been tested. DESIGN: This was a two-center, open-label study with a 4-week baseline, followed by 12 weeks of treatment with nefazodone at a median dose of 300 mg (mean, 303.66 +/- 65.57 mg; range, 100 to 450 mg depending on tolerability). Potential patients were required to report more than 15 days of headache per month for at least 3 months prior to screening. Only patients with at least 15 days of recorded headache during baseline were included in the final sample (N=52). Most patients (n=48) had a history of migraine based on International Headache Society criteria; 4 had primarily chronic tension-type headache, but with more migrainous features than permitted by International Headache Society criteria for a primary chronic tension-type headache diagnosis. RESULTS: Significant improvement was demonstrated for all headache diary measures, with significance levels ranging from P<.00001 for average intensity, duration, headache index (intensity x duration), peak intensity, headache days per week, and peak impairment, to P<.0033 for severe headache days per week, and P<.0051 for rescue medication days. During the last month of treatment, 71% of the patients completing the study showed at least a 50% reduction in headache index compared to baseline, and 59% had at least a 75% improvement. Visual analog scales completed at 4-week intervals showed significant improvement in patient ratings of overall headache status, quality of life, sleep, mood (P<.00001), and sexual function (P<.00053). Significant improvements were also observed in the Pain Disability Index (P<.00007), Beck Depression Inventory-II (P<.00001), Hamilton Rating Scale for Depression (P<.0008), and Hamilton Psychiatric Rating Scale for Anxiety (P<.00007). Headache indices for patients in the top quartile on the depression and anxiety scales (clinical depression/anxiety) did not differ from the other patients during baseline. However, patients who were depressed or anxious showed significantly more improvement over the course of 12 weeks of treatment (P<.0006 or less for the depression scales, P<.026 for anxiety). Common mild to moderate adverse events reported by 10% or more of the patients included fatigue, nausea, dry mouth, dizziness, sleep disturbance, blurred vision, irritability/nervousness, and sedation. Only 5 of the 52 patients discontinued the study due to adverse events: headache (2 patients), and nausea, sleep disturbance, and a drugged feeling (1 patient each). CONCLUSIONS: These results provide preliminary support for the efficacy of nefazodone in the prophylaxis of chronic daily headache. In this sample, nefazodone was safe and generally well tolerated. Patient ratings of sexual function improved over the course of treatment, in contrast to what is generally observed with most antidepressants. Nefazodone may be particularly beneficial for patients with chronic daily headache and comorbid depression. Further research is indicated. PMID- 11380645 TI - Blood nitrite levels in patients with migraine during headache-free period. AB - OBJECTIVE: To investigate blood nitrite levels after migraine attacks and to assess whether or not the change in nitric oxide levels observed during acute migraine persist after the attacks. BACKGROUND: Involvement of nitric oxide has been suggested in the initiation of acute migraine. Recent studies have shown alteration in the platelet response and platelet nitrite levels during migraine attacks. METHODS: Patients with migraine with aura and patients without aura were included in the study. The study was conducted on 50 patients with migraine and 90 healthy controls. Blood from the patients was collected at least 7 +/- 0.8 days after the last attack of migraine. Nitrite levels in the polymorphonuclear leukocytes, platelets, and plasma were estimated. Platelet aggregation response in some of these patients was also studied. RESULTS: No significant change in the polymorphonuclear leukocyte, platelet, and plasma nitrite levels in patients with migraine compared to controls was observed. Patients with migraine with aura had significantly lower polymorphonuclear leukocyte nitrite levels compared to those without aura (P<.05). In addition, no significant difference in the adenosine diphosphate-induced platelet aggregation was observed in the migraineurs compared to the healthy controls. CONCLUSIONS: Results obtained indicate that the platelet aggregation response and the blood nitrite levels were not altered significantly after an attack in the patients with migraine. PMID- 11380646 TI - Tension-type headache and stressful events: the role of selective memory in the reporting of stressors. AB - This study examined evidence for the role of selective memory for stressful events in women with headache. Previous studies have reported that on retrospective measures of stress, those with tension-type headache report more stressful events and rate the events as more stressful than did headache-free controls. However, when ratings are made concurrently, participants with headache and controls did not differ in their ratings of equivalent stressors presented in the laboratory. One theory for why differences are found in stress ratings made retrospectively, but not concurrently, is that selective memory biases recollection of past events in patients with headache. This study compared self report ratings of stressful events and their perceived impact made either concurrently or retrospectively to determine if selective memory might explain the discrepancies found in earlier studies. Participants included 20 patients with tension-type headache and 22 headache-free controls. Participants were compared on hourly, daily, and weekly measures of stressors and their perceived impact via hourly and nightly visual analog scale ratings, the Daily Stress Inventory, and the Weekly Stress Inventory. If support was to be offered to the hypothesis that selective memory biases the retrospective memory of patients with headache, then an interaction between group assignment (ie, headache versus control group) and time of rating would be expected. No such interaction occurred. Results from the study suggest that all participants have a tendency to overestimate stress on retrospective measures, but that patients with headache do not do so at a significantly different rate than do headache-free controls. The alternative hypothesis that patients with headache tend to appraise everyday events as more stressful is supported. PMID- 11380647 TI - Perceptions of pain in women with headache: a laboratory investigation of the influence of pain-related anxiety and fear. AB - The present study compared the responses of women with headache (chronic tension type, n = 27; migraine, n = 27) and controls (n = 27) to an acute pain laboratory task, the cold pressor test. Participants' pain perception (i.e., threshold and tolerance) and their fear/anxiety associated with pain were assessed during days 1, 2, or 3 of menses. Analyses pertaining to participants' responses to the cold pressor test (ie, pain threshold and tolerance) failed to show statistically significant group differences, even when covarying pain-related anxiety/fear. Analyses did, however, reveal significant group differences between migraineurs and controls in cognitive anxiety. Correlational analyses also revealed that cognitive anxiety, somatic anxiety, fear, and escape/avoidance were all significantly correlated with pain tolerance in the group with chronic tension type headache, but not in the other two groups. Subsequent multiple regressions, however, showed that the relationship between anxiety and pain tolerance was primarily a function of somatic anxiety. These results suggest that headache frequency plays a role in mediating the relationship between fear of pain and pain tolerance and that the models by Lethem and colleagues and McCracken may be relevant for understanding tension headache sufferers' responses to head pain. PMID- 11380648 TI - Medication-induced (analgesic rebound) headache: historical aspects and initial descriptions of the North American experience. AB - A review of the initial descriptions of medication-induced (misuse) headache in the North American literature indicates that this disorder was first identified in the mid-1950s. It was not until the early 1980s that this phenomenon became well established. PMID- 11380649 TI - Indirect costs of migraine in a public Brazilian hospital. AB - OBJECTIVES: To evaluate the indirect costs of migraine affecting employees of a public Brazilian hospital. BACKGROUND: Migraine is a common primary headache which has a negative influence on the well-being and quality of life, as well as the professional life, of affected individuals. METHODS: Our series consisted of 846 employees with migraine. The Lost Hours Equivalence Index, which considers both the hours lost due to the absence from work and reduction in productivity, was used to estimate the number of working hours lost due to migraine. RESULTS: Of the employees with migraine studied, 91% presented a mean 56.9% loss of productivity. The mean number of total lost working hours per month due to migraine was 6.5. The estimated total indirect cost of migraine was R $986 903.77 (US $815 622.54), implicating costs of R $241.30 (US $199.42) per employee per year. CONCLUSIONS: Based on its impact on life and the resulting costs, migraine should be considered a public health problem and thus measures should be adopted to reduce its impact on the individual and on society. PMID- 11380650 TI - Headache as only symptom in multiple cervical artery dissection. AB - We describe a patient with atypical headache as the only presenting symptom of spontaneous triple cervical artery dissection. As the patient suffered from arterial hypertension, a causative relation between headache and arterial hypertension was initially taken into consideration. However, four-vessel arteriography disclosed a dissection of both internal carotid arteries and the right vertebral artery. This unique case highlights the value of conventional arteriography for diagnosing cervical artery dissection. Since multiple cervical artery dissections are not rare, all cervical arteries should be examined by means of conventional arteriography when a dissection is suspected. PMID- 11380651 TI - Orgasm and migraine. PMID- 11380652 TI - Dubious about Depakote. PMID- 11380654 TI - Defining criteria of a putative inflammatory headache syndrome: cluster headache. PMID- 11380656 TI - Relationship between heterozygosity and asymmetry: a test across the distribution range. AB - The genetic basis of developmental stability, as measured by bilateral asymmetry, has been debated for over 50 years among developmental and evolutionary biologists. One of the central theories dealing with this relationship suggests that higher levels of genetic diversity, as reflected in heterozygosity, result in increased stability during development and thus in lower asymmetry. In this study, we aimed to test the relationship between asymmetry and heterozygosity at two levels: (1) the population level, where mean heterozygosity within a population is predicted to be negatively correlated with mean population asymmetry and (2) the individual level, where the proportion of heterozygous loci of an individual and its bilateral asymmetry estimates are predicted to be negatively correlated. While previous studies often focused on local populations, work across species ranges can answer the following questions. Are levels of heterozygosity correlated with levels of developmental instability, as estimated by bilateral asymmetry? Are patterns consistent across the distribution range, from the periphery towards the core? Does the relationship between genetic stress and bilateral asymmetry depend on the degree of environmental stress? We tested heterozygosity levels in 26 loci and asymmetry in third toe length in 11 populations of the chukar partridge (Alectoris chukar) across a sharp climatic gradient in Israel from the arid periphery, through the Mediterranean-desert ecotone towards the Mediterranean areas located further away from the range boundaries. Genetic diversity, as estimated using both observed and expected heterozygosity, was not associated with asymmetry at either the population or at the individual level. Whereas heterozygosity showed a hump-shaped pattern, peaking at the ecotone, asymmetry monotonically increased towards the range periphery. We argue that whereas asymmetry may serve as a useful tool for estimating changes in environmental stress, it may not be widely applicable for estimating genetic stress. PMID- 11380657 TI - Silent ribosomal cistrons are located at the pairing segment of the postreductional sex chromosomes of Apodemus sylvaticus (Rodentia, Muridae). AB - We analysed the karyotype of the rodent species Apodemus sylvaticus by G- and C banding, Ag-NOR-staining and in situ hybridization, with special attention to the sex chromosomes. NOR-bearing chromosome pairs were identified with simultaneous staining of G bands and NORs. In situ hybridization with an rDNA probe revealed the presence of silent ribosomal cistrons in both sex chromosomes. Studies of meiosis demonstrated that these inactive ribosomal cistrons are located in the pairing segment, which occupies the proximal portion of the telocentric sex chromosomes, and may thus be involved in postreduction caused by an obligatory chiasma in this position. PMID- 11380658 TI - The mating system of the fungus Cryphonectria parasitica: selfing and self incompatibility. AB - Although the genetic components of mating systems in fungi are well understood as laboratory phenomena, surprisingly little is known about their function in nature or about their role in determining mating patterns and population genetic structure. Our study of the mating system of the haploid ascomycete fungus, Cryphonectria parasitica, resulted in the following. (1) Laboratory crosses among 20 isolates, chosen randomly from North America and China, resolved into two incompatibility groups (occurring on both continents), confirming that C. parasitica has a diallelic, bipolar sexual self-incompatibility system, typical of other self-incompatible Ascomycetes, in which mating is only successful between isolates of opposite mating type. (2) PCR-based markers for mating-type alleles correlated perfectly with mating-type phenotypes of individual isolates. (3) Three genotypes, isolated from natural populations in Virginia and West Virginia, were inoculated onto chestnut trees in two sites in West Virginia and were confirmed to have self-fertilized and outcrossed in both sites. (4) Ten isolates, of a total of over 200 assayed, were confirmed to have self-fertilized in the laboratory, albeit at very low frequency. Five of these 10 isolates were ramets of a single genet, suggesting a genetic basis underlying the proclivity to self-fertilize in the laboratory. (5) Self-fertilization could not be induced in the laboratory with exudates (ostensibly containing pheromones) from isolates of opposite mating type. These results demonstrate that, a sexual self incompatibility system notwithstanding, self-fertilization occurs under both laboratory and field conditions in C. parasitica. The disparity between observations of frequent selfing in nature and rare selfing in the laboratory suggests that the mating system is under ecological as well as genetic control. PMID- 11380659 TI - The contrasting genetic architecture of wing size and shape in Drosophila melanogaster. AB - Surprisingly little is known about the genetic architecture of body size in natural populations of Drosophila melanogaster. Using both generation means and triple-test-cross analyses, we investigated the genetic architecture of wing size (an indicator of body size) and wing shape in a naturally occurring body size cline. For wing size, we found significant epistatic genetic variance and evidence of past directional selection for increased body size. While wing shape also exhibits significant epistatic genetic variance, there was no indication of directional selection, suggesting instead a history of optimizing selection. Our results support the idea that epistatic variance may be more common in natural populations than was once suspected. Also, our results suggest substantial directional selection on wing size but not shape. PMID- 11380660 TI - Temporal changes in allele frequency, genetic variation and inbreeding depression in small populations of the guppy, Poecilia reticulata. AB - We established three closed lines of N = 10 for the guppy Poecilia reticulata, to evaluate the relationships among temporal changes in allele frequency, genetic variation and inbreeding depression for a fitness-related trait in small populations. Genetic variation at the allozyme loci, expressed by the proportion of polymorphic loci, number of alleles per locus and heterozygosity, decreased somewhat in two closed lines but it increased in one closed line over six generations. Effective population size (Ne) at each generation was estimated from the standardized variance in the allele frequencies. The average Ne was 24.4, 10.3 and 10.0 in the three closed lines. The inbreeding coefficient calculated from the Ne increased to 0.186, 0.321 and 0.414, respectively. As an index of the amount of inbreeding depression, changes in salinity tolerance were examined, because this trait is strongly sensitive to inbreeding depression and decreases linearly with an increase in inbreeding coefficient. The mean value of the salinity tolerance significantly decreased to 82.5%, 71.7% and 67.6% in the three closed lines during the six generations, suggesting inbreeding depression for salinity tolerance. Although a significant correlation was not observed between the amount of inbreeding depression and the genetic variation, the amount of inbreeding depression correlated with the inbreeding coefficient calculated from Ne. The regression line indicated an 8.4% decrease in the mean per 10% increase in the inbreeding coefficient and was similar to that obtained directly from full sib matings. These results indicate that the temporal changes in the allele frequencies can provide an estimation of the amount of inbreeding depression during successive generations in small populations. PMID- 11380661 TI - Two male-killing Wolbachia strains coexist within a population of the butterfly Acraea encedon. AB - Inherited bacteria that kill male hosts early in their development are known from five insect orders. We ask to what extent the incidence of male-killers might be restricted by the rate at which new host-parasite interactions arise, by testing whether multiple male-killers have invaded a single host species. In Uganda, the butterflies Acraea encedon and A. encedana are both infected by the same strain of male-killing Wolbachia and there was no evidence of variation within the population. In Tanzanian A. encedon however, two phylogenetically distinct strains of male-killing Wolbachia were found within the same population. If this pattern of male-killer polymorphism is found to be general across infected species, it suggests that new male-killing infections arise frequently on an evolutionary time scale. Whether this polymorphism is stable, and what forces may be maintaining it, are unknown. PMID- 11380662 TI - Androgenesis from Festuca pratensis x Lolium multiflorum amphidiploid cultivars in order to select and stabilize rare gene combinations for grass breeding. AB - Androgenesis using amphidiploid cultivars of Festuca pratensis x Lolium multiflorum as parents, overcame earlier problems that gave rise to widespread plant sterility amongst androgenic Festulolium populations. Two Festuca pratensis x Lolium multiflorum (2n = 4x = 28) cultivars, Sulino and Felopa, were highly amenable to androgenesis and 10% of plants, including some novel androgenic genotypes, had sufficient fertility to produce progeny and further generations. The genomes of amphidiploid cultivars, which represent the F8 generation, were the result of considerable intergeneric chromosome recombination. Moreover, during cultivar development, natural and breeders' selection pressures had led to the assembly of gene combinations that conferred good growth characters and fertility with the removal of putative deleterious gene combinations. Over 80% of the androgenic plants derived from the amphidiploid F. pratensis x L. multiflorum (2n = 4x = 28) had 14 chromosomes and were likely to be dihaploids with a single genome of Lolium and of Festuca. In contrast, hybrids of F. pratensis x L. multiflorum (2n = 2x = 14) found naturally are invariably sterile. Structural reorganization within the genomes of the androgenic Festulolium plants had restored fertility in genotypes expected to contain the haploid genome of Lolium and Festuca. This provided opportunities for their future incorporation in breeding programmes and the development of fertile diploid Lolium-Festuca hybrids. Amongst the androgenic plants, Festulolium genotypes were recovered that conferred excellent drought resistance or freezing tolerance and were thought to be highly suitable for entry into plant breeding programmes. PMID- 11380663 TI - Polyandry in Lepidoptera: a heritable trait in Spodoptera exigua Hubner. AB - The genetic basis as well as the mode of inheritance of polyandry in Spodoptera exigua Hubner was studied in the laboratory by using a simple divergent selection experiment followed by F1 reciprocal crosses, F2 and backcrosses. There was an effective response to artificial selection for high (H line) and low (L line) female mating frequency with significant separation of the lines by the second generation of selection. The mean female mating frequency in the parental generation (1.57 matings per female) reached plateaus of 2.50 and 1.25 matings per female in the H and L lines, respectively, after six generations of selection. Selection response becomes saturated at about 90% and 25% levels of polyandry (percentage females re-mating) in the H and L lines, respectively, and consequently mono- and polyandric pure strains were not obtained. Polyandry levels in offspring from the H and L lines and their hybrids in F1, F2 and backcrosses consistently indicate that female mating frequency was more or less proportional to the relative amounts of genes derived from the H and L lines. Such a clear pattern of hybrid responses, together with the gradual selective changes under artificial selection, suggests the involvement of a polygenic system. Female mating frequencies from progeny of the two reciprocal F1 crosses were not significantly different, which suggest that the trait was autosomally inherited. Moreover, female mating frequency of F1 (pooled) progeny was not significantly different from the mid-parental value, which suggest no dominance. The computation of the Cavalli's joint scaling test consistently confirmed these results yielding values of d = 0.51 +/- 0.10 and h = 0.12 +/- 0.21. The broad sense heritability estimate was H2 = 0.73. It is concluded that polyandry in S. exigua is a polygenic, autosomal heritable trait and that additive genetic variance is available for selection for female mating frequency. The implications of the genetic basis of polyandry are briefly discussed in the context of current theories about this crucial insect mating system. PMID- 11380664 TI - Inbreeding depression in two populations of Arenaria uniflora (Caryophyllaceae) with contrasting mating systems. AB - I used parallel family-structured crossing designs to investigate the relative performance of self and outcross progeny in selfing and predominantly outcrossing populations of the annual plant Arenaria uniflora. The selfer population experienced much lower inbreeding depression (delta = 0.05 +/- 0.02 SE) than the outcrossers (delta = 0.19 +/- 0.02 SE). The negative association between genetic load and selfing rate suggests that purgable partially recessive alleles are the primary source of inbreeding depression, as does its late expression in both populations. Inbreeding depression in the selfer population, which naturally consists of highly inbred lines, was used to calculate the mean dominance (h = 0.33) and incidence rate (U = 0.30) of deleterious mutations. In the outcrosser population, significant variation among individuals in the expression of inbreeding depression may reflect lineage-specific differences in inbreeding history or, more probably, random variation in mutational load. The low (<< 0.5) inbreeding depression of outcrossers suggests that the maintenance of a mixed mating system in some A. uniflora populations and the evolution of nearly cleistogamous self-pollination in others may reflect local pollinator-mediated selection for selfing rather than the constant 3:2 genetic advantage invoked by many models. PMID- 11380665 TI - Roles of lineage sorting and phylogenetic relationship in the genetic diversity at the self-incompatibility locus of Solanaceae. AB - Allelic polymorphism at the S locus that determines the gametophytic self incompatibility (GSI) system in the pistil predates speciation. Understanding the evolution of a GSI system therefore requires knowledge of how lineage sorting and interspecific phylogenetic relationship affect S allele polymorphism. In searching for patterns of lineage sorting among species of various phylogenetic relationships, 22 S-alleles from 34 genets randomly taken at three Tennessee sites from a newly known GSI species Physalis longifolia were sequenced. Analyses of these data along with the previous sequences of three solanaceous species indicate that much of the combined allelic genealogy may be explained by lineage sorting and phylogenetic relationship. Using the mean terminal branch lengths of trans-specific alleles on the allelic genealogy to infer phylogenetic relationship among species, P. longifolia was found to be more closely related to P. cinerascens than to P. crassifolia. Nonetheless, the distribution of terminal branch lengths of P. longifolia was more similar to that of P. crassifolia than to that of P. cinerascens, suggesting phylogenetic relationship may have little effect on species-specific polymorphism. Similar habitat and growth characters, yet contrasting S-polymorphism, between P. longifolia and P. cinerascens also reject previous hypotheses that habitat and growth characters are the major factors responsible for interspecific differences in S-polymorphism. A likely scenario is that species-specific S-polymorphism is based on lineage sorting whose effect is further modified by species age and historical changes in population parameters. PMID- 11380666 TI - P elements and P-M characteristics in natural populations of Drosophila melanogaster in the southernmost islands of Japan and in Taiwan. AB - In order to study P element dynamics in natural populations of Drosophila melanogaster, 126 isofemale lines were examined from seven of the southernmost islands of Japan (the Sakishima Islands) and from Taiwan. Gonadal dysgenesis (GD) tests showed large divergences in the P-M phenotypes (P inducing and P repressing abilities) between the island populations. The P-M characteristics of each population, however, had not greatly changed in the past 15 years. Their genomic P element profiles are highly similar, consisting mostly of full-size P and of KP elements. We found no clear relationship between phenotype and genomic P element composition. PMID- 11380667 TI - Kin-structured subpopulations in Eurasian perch (Perca fluviatilis L.). AB - Based on ecological and behavioural studies it has been assumed that Eurasian perch (Perca fluviatilis) within one lake may not represent one panmictic population, but that they are subdivided into subpopulations. In order to investigate the genetic substructuring of populations, we used gene frequencies of five microsatellite loci to compare perch from six different sites from Lake Constance, Germany, and as outgroups perch from the lake Grosser Vatersee, Berlin, and two Swiss lakes, Lake Zurich and Lake Walensee. We examined whether homing behaviour of subadults to the spawning sites of their parents occurs and whether philopatric behaviour of adults results in significant population genetic substructuring. The distribution of genetic variation revealed two major, genetically distinct populations in Lake Constance: one in the eastern part of the lake and another in the western part (GST = 0.07). Within each of these two populations, no further genetic substructuring, nor any indication of inbreeding could be detected, either because genetic exchange was sufficiently high or because the time since separation has been too short. Homing behaviour of subadults to parental spawning sites after having spent several weeks of their life cycle in the pelagic zone could not be detected. Instead, subadults stay within either the western or the eastern region of the lake. There is evidence that some shoals contain full- and half-sibs. Despite females spawning in close proximity to each other, some siblings stay together. This might suggest that perch possess kin preferences and kin recognition. PMID- 11380668 TI - Low levels of genetic differentiation among populations of the freshwater fish Hypseleotris compressa (Gobiidae: Eleotridinae): implications for its biology, population connectivity and history. AB - The isolating nature of freshwater systems may lead to expectations of substantial genetic subdivision among populations of obligate freshwater species. We examined the genetic structure of populations of the freshwater fish Hypseleotris compressa (Gobiidae) using allozyme and mtDNA markers. Fifteen east coast Queensland populations and one Northern Territory population were sampled to examine levels of differentiation within and between drainages at near, medium and broad scales. Initial allozyme data suggested high levels of gene flow and connectivity among populations at broad spatial scales. However there was no significant relationship between geographical distance and gene flow among east coast populations which may indicate, among other possibilities, that these populations are not at equilibrium between gene flow and genetic drift. Analyses of a 567-bp fragment of the ATPase6 mtDNA gene revealed a star-shaped phylogeny, with many singleton, recently derived haplotypes. Tajima's test of neutrality was significantly negative. The allozyme and mtDNA data may be indicative of an historical demographic change that was reflected in the nonequilibrium pattern exhibited by contemporary populations. As estimating current levels of gene flow would violate basic assumptions of underlying models, approximations were not made. Nevertheless, patterns of genetic variation among populations of H. compressa do not match traditional expectations for a freshwater fish, and it would appear that there has been at least historical connectivity between populations now inhabiting different drainages. PMID- 11380669 TI - Interspecific and geographical variation in the sequence of rDNA expansion segment D3 of Ixodes ticks (Acari: Ixodidae). AB - The base sequence of the rDNA D3 expansion segment and flanking H14 stem varies between six species of Ixodes ticks (Acari: Ixodidae) where only 33 invariant sites occur among sequences of 123-203 bases in length. Multiple copies of D3 were sequenced from localities across the geographical ranges of four species to investigate deep population genetic structure. Two species, I. pacificus, from western North America, and I. ricinus, from Europe, have no sequence variation indicating a lack of deep genetic structure. One species, I. scapularis, from eastern North America has two forms of the D3 sequence that are distributed differently among northern vs. southern populations, suggesting recent divergence and hybridization. I. persulcatus, from Eurasia, has sequence variation between localities of the order of that observed between other species, suggesting a long history of population isolation and deep genetic structure. With the exception of I. scapularis, sequence variation was not observed within localities. This indicates that cellular processes underpinning concerted evolution have homogenized populations and species for particular rDNA sequence variants. PMID- 11380670 TI - Lack of genetic differentiation among widely spaced subpopulations of a butterfly with home range behaviour. AB - We examine seven geographically separate subpopulations of Heliconius charithonia, a butterfly with well-documented home range behaviour, in Miami-Dade County, Florida, for genetic differentiation using cellulose acetate electrophoresis. These subpopulations exhibit little genetic variation (percent polymorphic loci = 27, average heterozygosity = 0.103) especially in comparison to populations of the same and related species from mainland South America. Allele frequencies do not differ among the subpopulations in south Florida and estimates of Wright's fixation index (FST) support that there is no detectable genetic differentiation among them. This result supports an earlier finding that the dispersal ability of Heliconius butterflies may be underestimated. However, it is unlikely that increased dispersal ability alone could account for the lack of genetic differentiation observed among subpopulations separated by almost 80 km. Given the likely effective population size of these subpopulations (Ne = 205) and the average generation time of this species in the subtropics (in the range of 30-90 days), this lack of genetic differentiation is best explained by current or very recent gene flow following a stepping-stone model. Furthermore, this result provides evidence that the current extensive degree of habitat fragmentation surrounding the city of Miami does not limit gene flow among urban subpopulations of Heliconius charithonia. PMID- 11380671 TI - Burnet oration. T-cell survival and the role of cytokines. AB - Typical T cells are long-lived resting cells. Despite their quiescent appearance, there is increasing evidence that T cells are subjected to continuous stimulation through contact with various stimuli, notably by self peptide/MHC complexes and cytokines. These stimuli keep T cells alive and also cause intermittent entry into cell cycle. PMID- 11380672 TI - Evaluation of the T cells and costimulatory molecules in the protective efficacy of 30 kDa secretory protein against experimental tuberculosis. AB - Phenotypic changes of T lymphocytes and B7 costimulatory molecules in mice first vaccinated with mycobacterial 30 kDa secretory protein and then challenged with Mycobacterium tuberculosis H37Rv (Group 2) were monitored using flow cytometry and compared with non-vaccinated, but challenged mice (Group 1). In Group 1, the proportion of CD3+ and CD4+ T cells increased until 28 days postinfection (p.i.) and then declined to levels even less than healthy controls (non-vaccinated and non-challenged healthy mice), especially at later stages of infection (i.e. 72 days p.i.). However, the levels of CD8+ T cells did not decline and remained either significantly higher or similar to healthy control levels. In Group 2, however, the levels of CD3+ and CD4+ T cells did not decline as seen in Group 1, but remained significantly higher than in Group 1. Furthermore, the profile of CD8+ T cells remained similar to what was observed in Group 2. In order to elucidate Th1-Th2 bias, the ratio of IgG2a/IgG1 bearing cells was enumerated by flow cytometry. A predominantly Th1 response was observed in Group 1 until 28 days p.i. (IgG2a/IgG1 ratio was >1). However, in Group 2 a predominantly Th1 bias was observed throughout the period studied in terms of IgG2a/IgG1 ratios. The examination of expression of B7-1 and B7-2 costimulatory molecules on a monocyte gated population was carried out. In Group 2, the B7-1 and B7-2 expression was found to be significantly higher compared to Group 1, especially at later stages of infection (i.e. 60 and 72 days p.i.). Thus, these results suggest the capability of mycobacterial 30 kDa secretory protein in restoring the T-cell responses, especially at later stages of infection, possibly by augmentation of both B7-1 and B7-2. Further, these costimulatory molecules are probably required for effective T-cell responses against virulent mycobacterial challenge in a murine model of tuberculosis. PMID- 11380673 TI - Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) for measurement of cytokine and growth factor mRNA expression with fluorogenic probes or SYBR Green I. AB - Real-time quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) is the method of choice for rapid and reproducible measurements of cytokine or growth factor expression in small samples. Fluorescence detection methods for monitoring real-time PCR include fluorogenic probes labelled with reporter and quencher dyes, such as Taqman probes or Molecular Beacons and the dsDNA-binding dye SYBR Green I. Fluorogenic (Taqman) probes for a range of human and rat cytokines and growth factors were tested for sensitivity and compared with an assay for SYBR Green I quantification using real-time fluorescence monitoring (PE Applied Biosystems Model 7700 sequence detector). SYBR Green I detection involved analysis of the melting temperature of the PCR product and measurement of fluorescence at the optimum temperature. Fluorogenic probes provided sensitive and reproducible detection of targets that ranged from low (<10 copies/reaction) to high (>107 copies/ reaction) expression. SYBR Green I gave reproducible quantification when the target gene was expressed at moderate to high levels (> or =1000 copies/reaction), but did not give consistently reproducible quantification when the target gene was expressed at low levels. Although optimization of melting temperature improved the specificity of SYBR Green I detection, in our hands it did not equal the reproducible sensitivity and specificity of fluorogenic probes. The latter method is the first choice for measurement of low-level gene expression, although SYBR Green I is a simple and reproducible means to quantify genes that are expressed at moderate to high levels. PMID- 11380674 TI - In vitro and in vivo expression of a nephritogenic Ig heavy chain determinant: pathogenic autoreactivity requires permissive light chains. AB - Lymphocyte antigen receptors are promising targets for immune intervention strategies in disorders marked by repertoire skewing or expansion of lymphocyte subsets. Appropriate application of immune receptor modulation is predicated on understanding the role of a particular receptor in pathogenesis and disease regulation. The VHB/W16 gene, restricted to mice carrying the j haplotype for the J558 family, is overexpressed by murine lupus anti-DNA Ig. This gene is also expressed recurrently among nephritogenic anti-DNA Ig recovered from several autoimmune strains, suggesting that cells expressing this pathogenic receptor are positively selected during disease progression. To explore the extent and mechanisms by which Ig H chains expressing this gene contribute to autoimmunity, an Ig H chain gene was engineered for in vitro and in vivo recombination studies. Site-directed mutagenesis generated unique restriction sites to link PCR amplified V region (VDJ) cDNA to previously isolated genomic fragments containing Ig regulatory and signal sequences. The new 3 kb VDJ gene was then ligated to a 9 kb fragment encoding the IgM constant region. Transfection of H chain loss variant myeloma with the complete 12 kb construct, termed 238H-Cmicro, resulted in secretion of intact Ig pairing 238H-Cmicro, with a lambda L chain; however, transfectant Ig lacked autoreactivity and pathogenicity. Introduction of the 238H Cmicro H chain as a transgene onto the non-autoimmune C57BL/6 background resulted in abundant B cell surface expression of 238H-Cmicro, however, four transgenic Ig recovered by fusion of LPS-stimulated splenocytes and formed by combination of 238H-Cmicro, with endogenous kappa chains do not bind DNA or laminin. These results indicate that the antigen binding sites encoded by this disease associated gene and/or H chain must associate with permissive L chains to specify autoimmunity. The 238H-Cmicro, transgenic model should prove useful in dissecting the in vivo fate of 238H-Cmicro, L combinations that produce pathogenic autoreactive receptors and in evaluating receptor-targeted interventions. PMID- 11380675 TI - Complement and myoblast transfer therapy: donor myoblast survival is enhanced following depletion of host complement C3 using cobra venom factor, but not in the absence of C5. AB - Myoblast transfer therapy (MTT) is a potential cell therapy for myopathies such as Duchenne Muscular Dystrophy and involves the injection of cultured muscle precursor cells ('myoblasts') isolated from normal donor skeletal muscles into dystrophic host muscle. The failure of donor myoblast survival following MTT is widely accepted as being due to the immune response of the host. The role of complement as one possible mechanism for the initial, very rapid death of myoblasts following MTT was investigated. Donor male myoblasts were injected into the tibialis anterior (TA) muscles of female host mice that were: (i) untreated; (ii) depleted of C3 complement (24 h prior to MTT) using cobra venom factor (CVF); and/or (iii) deficient in C5 complement. Quantification of surviving male donor myoblast DNA was performed using the Y-chromosome specific (Y1) probe on slot blots for samples taken at 0 h, 1 h, 24 h, 1 week and 3 weeks after MTT. Peripheral depletion of C3 was confirmed using double immunodiffusion, and local depletion of C3 in host TA muscles was confirmed by immunostaining of muscle samples. Cobra venom factor treatment significantly increased the initial survival of donor myoblasts, but there was a marked decline in myoblast numbers after 1 h and little long-term benefit by 3 weeks. Strain specific variation in the immediate survival of donor male myoblasts following MTT in untreated C57BL/10Sn, DBA-1 and DBA-2 (C5-deficient) female hosts was observed. Cobra venom factor depletion of C3 increased initial donor male myoblast survival (approximately twofold at 0 h) in C57BL/10Sn and DBA-1 host mice and approximately threefold in DBA-2 hosts at 0 h and 1 h after MTT. The rapid and extensive number (approximately 90%) of donor male myoblasts in untreated DBA-2 mice (that lack C5) indicates that activation of the membrane attack complex (MAC) plays no role in this massive initial cell death. The observation that myoblast survival was increased in all mice treated with CVF suggests that CVF may indirectly enhance donor myoblast survival by a mechanism possibly involving activated C3 fragments. PMID- 11380676 TI - Expression of cell surface adhesion molecules by peripheral blood eosinophils during Trichostrongylus colubriformis infection in sheep. AB - The effect of infection of sheep with the gastrointestinal nematode parasite Trichostrongylus colubriformis on expression of adhesion molecules CD11a, CD11b, CD11c, CD18, CD44, CD49d and CD62L by peripheral blood eosinophils was examined by flow cytometry. Initially, to establish the sensitivity of adhesion molecules to inflammatory signals, eosinophil-rich exudates were elicited in non-lactating mammary glands of immune sheep by infusion of 50 microg of soluble antigen extract from T. colubriformis third stage larvae. Eosinophils comprised 40.8% of mammary leucocytes and 4.5% of peripheral blood leucocytes. In comparison with blood, the percentage of eosinophils expressing CD18 increased and the percentage expressing CD62L decreased in exudates and the mean fluorescent intensity, an indicator of receptor number per cell, for CD11a and CD49d also decreased on exudate eosinophils. Peripheral blood eosinophils were examined over 8 weeks during trickle infection of immune sheep with infective or irradiated third stage larvae of T. colubriformis. During the last 3 weeks of infection, CD11a staining decreased in infected sheep and CD44 staining decreased in sheep receiving either infective or irradiated larvae. Other surface markers did not change. The results indicate that systemic changes in expression of adhesion molecules by eosinophils occur during T. colubriformis infection in sheep. PMID- 11380677 TI - Peptide immunization of guinea pigs against Chlamydia psittaci (GPIC agent) infection induces good vaginal secretion antibody response, in vitro neutralization and partial protection against live challenge. AB - Immunization of female guinea pigs with a chimeric peptide consisting of variable domain IV (VDIV) and a region known as GP8 from the major outer membrane protein of Chlamydophila caviae, formerly Chlamydia psittaci guinea pig inclusion conjunctivitis strain, was performed to assess whether humoral immune responses could be elicited in the reproductive tracts of immunized animals. The C. caviae strain is able to cause a sexually transmitted infection in the guinea pig that closely parallels C. trachomatis infections in humans. The best anti-VDIV antibody response in vaginal secretions was achieved by intraperitoneal priming with subsequent intravaginal boosting (P < 0.001). Dot-blot analyses of vaginal secretions confirmed that these anti-VDIV antibodies, produced against a linear peptide, were able to recognize and bind to whole conformational C. caviae elementary bodies. Following live intravaginal challenge with C. caviae, a significant reduction in the intensity (P = 0.01) and an apparent reduction in the duration of the infection was evident between the guinea pigs immunized with VDIV-GP8 and non-immunized controls. PMID- 11380678 TI - Oxidative burst and metallothionein as a scavenger in macrophages. AB - The role of metallothionein (MT) in the scavenging of superoxide radicals (*O2-) generated by macrophages has been examined. The present work has focused on the effects of added cadmium, a known inducer of MT biosynthesis, on determined amounts of superoxide radicals produced by in vitro cultured rat peritoneal macrophages on their stimulation with phorbol-12-myristate-13-acetate (PMA). The levels of superoxide radicals (*O2-) have been found to decrease when cadmium was added to cells exposed to PMA. However, substantially lower levels of MT have been determined in this case compared to cells untreated with PMA. This effect could be reversed by incubation of the PMA and cadmium-treated cells with a reducing agent, 2-mercaptoethanol (2-ME). Results suggest that *O2- caused thiolate oxidation and subsequent metal loss, thus reducing the cellular MT content as quantified by the silver saturation METHOD: This conclusion is supported by cell-free experiments in which the oxidation of rabbit MT-I by a xanthine/xanthine-oxidase system could be reversed by its subsequent reduction with 2-ME. The data presented provide direct evidence of the involvement of MT in scavenging superoxide radicals in living cells. PMID- 11380679 TI - Uptake of HIV and latex particles by fresh and cultured dendritic cells and monocytes. AB - Blood dendritic cells (DC) efficiently carry HIV-1 and transmit infection to CD4+ T cells in the absence of productive infection of the APC. Fluorescent latex beads were used to define the endocytic pathways that may contribute to this non infectious pathway of virus carriage. Beads between 14 nm and 2300 nm in diameter were taken up by uncultured blood DC, but uptake of beads larger than 280 nm was much reduced in the DC compared to monocytes. After culture, there was a reduction in bead carriage in DC compared to monocytes. In the DC, beads were found as small aggregates in class II containing compartments or as single beads just below the cell surface. Beads accumulated in monocytes as aggregates in class II negative compartments. Bead recycling occurred in DC, but not in the fresh or cultured monocytes. Electron microscopy of HIV-1-pulsed DC cultured with CD4+ T cells showed accumulation of apoptotic debris and virions within endosomes in the DC. The peripheral location and recycling of endocytosed material in DC provides a pathway for virion transfer from DC to T cells that does not occur in monocytes. PMID- 11380680 TI - Cell death mediated by alloreactive cytotoxic T cells via the granule exocytosis or the Fas pathway is independent of p34cdc2 kinase: Fas dependent killing of cells arrested in the cell cycle. AB - Inappropriate activation of p34cdc2 kinase has been shown to occur during apoptosis induced by cytotoxic T-cell derived perforin and fragmentin. We analysed the effect of two inhibitors of p34cdc2 kinase on alloreactive Tc-cell mediated lysis and DNA fragmentation of P815 and L1210 target cells. Olomoucine, a specific inhibitor of cyclin dependent kinases, did not affect DNA fragmentation in the target cells. Lysis of olomoucine-treated target cells as assessed by 51Cr release over a typical 8-h period was also unaffected. We also examined the effects of thapsigargin on target cell death. This toxin causes increased intracellular calcium rises that then result in irreversible inhibition of cyclin dependent kinases, including p34cdc2 kinase. The same extent of specific cell lysis was induced by cytotoxic T cells from perforin(-/-), granzyme B(-/-), granzyme A(-/-), perforin(-/-) X granzymeB(-/-) X granzymeA(-/-) KO mice or normal mice in untreated target cells or target cells treated with either olomoucine or thapsigargin. Similarly DNA fragmentation measured by release of tritiated DNA was also unaffected. Thus inhibition of p34cdc2 kinase affects neither the Fas nor the perforin/granzyme pathways of alloreactive cytotoxic T cell killing as measured by DNA fragmentation or chromium release. P815 cells treated with olomoucine were arrested in the cell cycle after 12-16 h exposure to the toxin. After cell cycle arrest, target cells now showed enhanced 51Cr release induced by effector cytotoxic T cells (CTL) derived from perforin(-/-) mice compared to untreated cells. This lysis was accompanied by an increase in cell surface Fas expression. Olomoucine induced cell cycle arrest and expression of Fas was reversible and when cells re-entered the cell cycle, surface expression of Fas was lost. PMID- 11380681 TI - A novel system for convenient detection of low-affinity receptor-ligand interactions: chelator-lipid liposomes engrafted with recombinant CD4 bind to cells expressing MHC class II. AB - The interactions of cell surface receptors with their ligands, crucial for initiating many immunological responses, are often stabilized by receptor dimerization/oligomerization, and by multimeric interactions between receptors on one cell with their ligands or cognate receptors on the apposing cell. Current techniques for studying receptor-ligand interactions, however, do not always allow receptors to move laterally to enable dimerization/ oligomerization, or to interact multimerically with ligands on cell surfaces. For these reasons detection of low- affinity receptor-ligand interactions has been difficult. Utilizing a novel chelator-lipid, nitrilotriacetic acid di-tetradecylamine (NTA DTDA), we have developed a convenient liposome system for directly detecting low affinity receptor-ligand interactions. Our studies using recombinant soluble forms of murine CD40 and B7.1, and murine and human CD4, each possessing a hexhistidine tag, showed that these proteins can be anchored or 'engrafted' directly onto fluorescently labelled liposomes via a metal-chelating linkage with NTA-DTDA, permitting them to undergo dimerization/oligomerization and multimeric binding with ligands on cells. Fluorescence- activated cell sorter (FACS) analyses demonstrated that while there is little if any binding of soluble forms of murine CD40 and B7.1, and murine and human CD4 to cells, engrafted liposomes bind specifically to cells expressing the appropriate cognate receptor, often giving a fluorescence 4-6-fold above control cells. Such liposomes could detect directly the low-affinity interaction of murine CD40 and B7.1 with CD154- and CD28-expressing cells, respectively, and the interaction of CD4 with MHC Class II, which has hitherto defied direct detection except through mutational analysis and mAb blocking studies. PMID- 11380682 TI - Serum prolactin is associated with apoptosis in men with human immunodeficiency virus infection. AB - We examined the in vivo and in vitro production of prolactin (PRL) in 20 untreated HIV-infected men compared to 14 uninfected men and its association with the cell cycle and apoptosis. Compared to uninfected men, the HIV-infected men had: (i) higher fasting serum bioactive (BIO) PRL; (ii) lower serum immunoreactive (RIA) and BIO-PRL responses to intravenous metoclopramide; (iii) greater BIO-RIA PRL ratio both fasting and during intravenous metoclopramide; (iv) lower percentage of non-stimulated PBMC in the G0/G1 phase, but a higher percentage in the S phase, of the cell cycle with normal response to Concanavalin A; and (v) higher in vitro production of BIO-PRL by non-stimulated PBMC, which was blocked after Concanavalin-A. Fasting serum BIO-PRL positively correlated with the percent of non-stimulated PBMC in S + G2/M phases. The percentage of apoptotic PBMC negatively correlated with CD4+ T lymphocytes and with the area under the serum RIA-PRL curve, but positively correlated with the area under the curve for the BIO/RIA ratio. These results suggest that in these HIV-infected men: (i) a diminished dopaminergic tone may exist, as an adaptive mechanism attempting to survive; and (ii) BIO-PRL may participate as a cofactor in the stimulation of T-cell proliferation. PMID- 11380683 TI - Identification of soluble transforming growth factor-beta receptor III (sTbetaIII) in rat milk. AB - Transforming growth factor-beta (TGF-beta) is present at high concentrations in maternal milk. In milk TGF-beta2 is the predominant isoform. For function TGF beta2 requires TbetaRIII to facilitate efficient binding to the TGF-beta receptor types I and II signalling complex. We have shown that TGF-beta receptor types I (TbetaRI), II (TbetaRII) and III (TbetaRIII) are coexpressed in the suckling rat intestine. Immunostaining for TbetaRIII was also observed in the intestinal lumen prior to weaning. TbetaRIII (or betaglycan) has been reported in serum, cell culture medium and extracellular matrix. To determine whether a soluble form of TbetaRIII is present in milk, the rat milk aqueous phase was analysed by slot blot and Western blot. Soluble TbetaRIII was detected in milk throughout lactation. Western blot analysis of rat milk revealed a high molecular weight band of glycosylated protein of >200 kDa, with a core protein of approximately 110-120 kDa that comigrated with recombinant TbetaRIII. Immunoabsorption of soluble TbetaRIII (sTbetaRIII) from milk resulted in partial depletion of active TGF-beta from milk, suggesting that the receptor may interact with ligand in milk. In addition rat pups suckled on mother's milk demonstrated an enhanced labelling of TbetaRIII in the gut, as compared with pups fed on a rat milk substitute (RMS). These findings suggest that milk sTbetaRIII is functional, and may modulate milk-derived TGF-beta function in the developing intestine. PMID- 11380684 TI - Fractalkine, a CX3C-chemokine, functions predominantly as an adhesion molecule in monocytic cell line THP-1. AB - A newly identified CX3C-chemokine, fractalkine, expressed on activated endothelial cells plays an important role in leucocyte adhesion and migration. Co immobilized fractalkine with fibronectin or intercellular adhesion molecule-1 enhanced adhesion of THP-1 cells, which express the fractalkine receptor (CX3CR1), compared with that observed for each alone. That adherence was fractalkine-dependent and was confirmed in blocking studies. However, soluble fractalkine induced little chemotaxis in THP-1 cells in comparison to monocyte chemotactic protein-1 (MCP-1), which induced a strong chemotactic response. Moreover, the membrane form of fractalkine expressed on ECV304 cells reduced MCP 1 mediated chemotaxis of THP-1 cells. These results indicate that fractalkine may function as an adhesion molecule between monocytes and endothelial cells rather than as a chemotactic factor. PMID- 11380685 TI - CD43 potentiates CD3-induced proliferation of murine intestinal intraepithelial lymphocytes. AB - The involvement of CD43 in cell proliferation of murine intestinal intraepithelial lymphocytes (IEL) has been studied in in vitro CD3-stimulated cell cultures. In the presence of either IL-2 or IL-15, CD3 stimulation of IEL resulted in low levels of proliferation as measured by thymidine incorporation, whereas no proliferation occurred upon CD3 stimulation in the absence of cytokines. The combination of both cytokines to IEL cultures synergistically enhanced CD3-induced proliferation by approximately threefold that of cultures supplemented with either cytokine alone. Most importantly, however, proliferation of IEL was significantly greater when CD3 stimulation occurred in conjunction with CD43 triggering, indicating that CD43 functions as a coactivational signal for murine IEL. These findings indicate that a spectrum of potential proliferative responses exist among murine IEL depending on the types and combinations of signals received, and that because under normal conditions murine IEL are largely devoid of CD28 expression, a classical T-cell coactivational molecule, the capacity for high-level IEL proliferation may reside with CD43. PMID- 11380686 TI - The many faces of host responses to tuberculosis. PMID- 11380687 TI - Normal and abnormal secretion by haemopoietic cells. AB - The secretory lysosomes found in haemopoietic cells provide a very efficient mechanism for delivering the effector proteins of many immune cells in response to antigen recognition. Although secretion shows some similarities to the secretion of specialized granules in other secretory cell types, some aspects of secretory lysosome release appear to be unique to melanocytes and cells of the haemopoietic lineage. Mast cells and platelets have provided excellent models for studying secretion, but recent advances in characterizing the immunological synapse allow a very fine dissection of the secretory process in T lymphocytes. These studies show that secretory lysosomes are secreted from the centre of the talin ring at the synapse. Proper secretion requires a series of Rab and cytoskeletal elements which play critical roles in the specialized secretion of lysosomes in haemopoietic cells. PMID- 11380688 TI - Role of oestrogen receptors alpha and beta in immune organ development and in oestrogen-mediated effects on thymus. AB - Oestrogens affect the development and regulation of the immune system. To determine the role of oestrogen receptors alpha (ER-alpha) and beta (ER-beta) on the development of the immune system, male ER-alpha (ERKO) and ER-beta (BERKO) mice, as well as alphabeta-double knockout (DERKO) mice, were studied. Deletion of ER-alpha led to hypoplasia of both thymus and spleen. Interestingly, a higher frequency of immature double CD4+ CD8+ thymocytes was found in ER-alpha(-) mice compared with ER-alpha(+) mice. Female oophorectomized BERKO mice given oestradiol (E2) displayed a similar degree of thymic atrophy compared with the wild-type strain but showed only limited involution of thymus cortex and no alteration of thymic CD4/CD8 phenotype expression. Our data demonstrate that expression of ER-alpha, but not ER-beta, is mandatory in males for development of full-size thymus and spleen, whereas expression of ER-beta is required for E2 mediated thymic cortex atrophy and thymocyte phenotype shift in females. A potential background for the above findings may be down-regulated activity in the growth hormone/insulin-like growth factor-1 (GH/IGF-1) axis in males lacking ER alpha and suppressed sensitivity of females lacking ER-beta to E2-mediated suppression of IGF-1. PMID- 11380689 TI - Early Vlambda diversification in sheep. AB - This study examined a number of tissues during early gestation in foetal sheep to determine the earliest site of Vlambda expression and time of generation of the Vlambda repertoire. Tissues, including spleen, liver, gut, blood and bone marrow, were obtained from 48, 55, 60 and 63 gestational day (g.d.) ovine foetuses and cDNA libraries were prepared from them by reverse transcription-polymerase chain reaction. Clones were randomly selected from cDNA libraries and subjected to sequencing. Analysis of these sequences and comparison with a pool of germline genes led to the following conclusions. The expression of Vlambda occurs earlier in spleen (48 g.d.) than in all of the other tissues examined. Also, diversity is seen earlier and at higher levels in early foetal spleen than in all of the other tissues examined. In this regard, it is notable that splenic Vlambda expression is readily apparent even before such gut-associated lymphoid tissue as the ileal Peyer's patch (IPP) has developed. Two germline Vlambda genes, 5.1 and 5.3 predominate in early immunoglobulin lambda light-chain gene rearrangement. Examination of Jlambda usage revealed the existence of a new Jlambda gene and its utilization during the early phases of the development of the ovine antibody repertoire. This study indicates that sites other than the IPP contribute to the diversification of the Vlambda repertoire in sheep. We suggest that it is likely that foetal spleen may provide a partially diversified B-cell repertoire before the IPP becomes active as a major site for massive clonal expansion and extensive diversification of B cells. PMID- 11380690 TI - Polymorphism of the human alpha1 immunoglobulin gene 3' enhancer hs1,2 and its relation to gene expression. AB - We studied the hs1,2 transcriptional enhancer identified downstream of the human alpha1 gene of the immunoglobulin H (IgH) locus, for which two different allelic configurations (a and b) were previously reported by Southern blotting. By using a polymerase chain reaction (PCR) method we amplified minisatellites within the hs1,2 core enhancer, with variable numbers of tandem repeats (VNTR) defining three 'PCR alleles' alpha1A, alpha1B and alpha1C (including one, two and three repeats, respectively). Five different alpha1 h1,2 genotypes were encountered in a population of 513 donors, representing 13.8, 34.5, 49.7, 1.3 and 0.6% for the AA, BB, AB, AC and BC genotypes, respectively. Luciferase assays showed that increasing the number of minisatellites increased the transcriptional strength of the alpha1 hs1,2 enhancer. Simultaneous determination of Southern blot alleles and VNTR alleles only showed a partial linkage between both types of polymorphism, altogether defining at least six different allelic forms of the 3'alpha1 region. In conclusion, the present study further demonstrates the genetic instability of the 3'alpha region, for which multiple alleles have been generated through inversions and internal deletions and/or duplications. This study also strengthens the hypothesis that the polymorphism at the IgH 3' regulatory region of the alpha1 gene could play a role in the outcome of diseases involving immunoglobulin secretion. PMID- 11380691 TI - Macrophages present exogenous antigens by class I major histocompatibility complex molecules via a secretory pathway as a consequence of interferon-gamma activation. AB - Macrophages can process and present exogenous antigens on major histocompatibility complex (MHC) class I molecules through an alternative mechanism involving the internalization of antigens and the secretion of peptides loading MHC class I molecules at the cell surface. In this paper, we found that interferon-gamma (IFN-gamma) -activated macrophages infected with Salmonella typhimurum secreted peptides able to load empty MHC Kb molecules on co-cultured TAP-2-deficient RMA-S cells, added as targets for peptide loading. The increase in class I Kb on the RMA-S cells, resulting from the macrophage-derived peptides, exhibited a comparable stability as the direct addition of an exogenous Kb binding peptide (OVA257-264) to the RMA-S cells. In both cases, the Kb complexes were stable for at least 3 hr after separating the RMA-S cells from the macrophages. The endosomal inhibitors, leupeptin and ammonium chloride, did not inhibit the release of peptides and the increase in Kb staining on the RMA-S cells in the co-culture systems. Brefeldin A also had no effect. P815 cells previously co-cultured with Salmonella-infected macrophages became targets for cytotoxic T lymphocytes isolated from Salmonella-infected BALB/c mice. Taken together, our data suggest that IFN-gamma-activated macrophages process exogenous antigens in an intracellular compartment where serine proteases generate peptides released to the external environment for loading empty MHC class I molecules at the cell surface. This TAP-independent mechanism for the MHC class I presentation may be involved in priming cytotoxic T lymphocytes against intracellular pathogens in vivo. PMID- 11380692 TI - Immunostimulation by the synthetic lipopeptide P3CSK4: TLR4-independent activation of the ERK1/2 signal transduction pathway in macrophages. AB - Synthetic lipopeptides based on bacterial lipoprotein are efficient activators for monocytes/macrophages inducing the release of interleukin (IL)-1, IL-6, tumour necrosis factor-alpha (TNF-alpha), reactive oxygen/nitrogen intermediates, and the translocation of nuclear factor kappaB (NFkappaB). In this report we investigate the signal transduction pathways involved in leucocyte activation by the synthetic lipopeptide N-palmitoyl-S-[2,3-bis(palmitoyloxy)-(2R,S)-propyl]-(R) cysteinyl-seryl-(lysyl)3-lysine (P3CSK4). We show that P3CSK4 activates mitogen activated protein (MAP)-kinases ERK1/2 and MAP kinase (MAPK)-kinases MEK1/2 in bone-marrow-derived macrophages (BMDM) and in the macrophage cell line RAW 264.7. Additionally, we could detect differences between the P3CSK4 and lipopolysaccharide (LPS)-induced phosphorylation of MAP kinases: Different levels in phosphorylation were found both in kinetics and dose-response using RAW 264.7 cells or BMDM from BALB/c and LPS responder mice (C57BL/10ScSn) or LPS non responder mice (C57BL/10ScCr). The lipopeptide activated the MAPK-signalling cascade in both LPS responder and non-responder macrophages, whereas LPS induced the MAPK signalling pathway only in macrophages derived from LPS responder mice. An approximately 70% decrease of lipopeptide induced NFkappaB translocation and an about 50% reduction of nitric oxide (NO) release was observed in the presence of anti-CD14. These data correspond to the reduction of phosphorylation of ERK1/2 after stimulation with P3CSK4 in the presence of anti-CD14 antibodies. Inhibition of MEK1/2 by PD98059 completely reduced the lipopeptide-induced phosphorylation of ERK1/2 indicating that MEK1/2 are solely responsible for the phosphorylation of the downstream-located MAP kinases ERK1/2. PMID- 11380693 TI - Acquisition of immune function during the development of the Langerhans cell network in neonatal mice. AB - The immunological function of the Langerhans cell (LC) network in neonatal skin was examined by defining the development of cutaneous immunity relative to the structure, phenotype and function of the epidermal LC network in neonatal, juvenile and adult mice. Analysis of epidermal sheets showed the presence of major histocompatibility complex (MHC) II+, multilectin receptor DEC-205- cells within the epidermis of 3-day-old mice; both cell density and DEC-205 expression increased until day 14. When visualized with antibodies directed at MHC II, the network was poorly formed in 3- and 7-day-old mice, as there was a lower cell density and poor MHC II expression on dendritic processes, compared to mice at day14. Application of a fluorescent antigen to 3-day-old mice revealed that the LC were inefficient in transporting antigen to the draining lymph node. There was an improvement at day 7 and by day 14 comparable numbers of antigen carrying cells were detected in the lymph nodes of 6-week-old mice. The reduced antigen carriage in 3- and 7-day-old mice correlated with a poor contact sensitivity response. This was not simply due to failure to present antigen, but development of immunosuppression, as transfer of T cells from adult mice that were previously treated with antigen when they were 3 days old, to adult recipients resulted in antigen specific immunosuppression. Analysis of CD80 and CD86 expression showed that LC from day 3 skin expressed CD80, but not CD86 and application of antigen through this skin was inefficient in upregulating CD86. These findings indicate that when the neonatal LC network is poorly developed it is functionally immature and antigen applied through this 'functionally immature network' results in antigen specific immunosuppression. PMID- 11380694 TI - Differential endocytotic characteristics of a novel human B/DC cell line HBM Noda: effective macropinocytic and phagocytic function rather than scavenging function. AB - In order to characterize a novel human B cell-lineage dendritic cell line (B/DC line) as an antigen-presenting cell (APC), we compared three types of endocytosis (micropinocytosis via a clathrin-coated pit, macropinocytosis via membrane ruffling, and phagocytosis) among myeloid-related, macrophage (Mphi) cell lines and a B/DC line. In the present examination, we used a unique human dendritic cell (DC) line, HBM-Noda (Noda). Flow cytometric and immunocytochemical analyses revealed that Noda not only expresses some DC markers, but also it expresses some B-cell associated markers. Noda shows strong capacities to stimulate allogenic T cells, to produce immunoglobulin G (IgG), and to perform immunoglobulin gene rearrangement. These data strongly suggest that Noda is a B-cell lineage DC line. The endocytic differences among these cell lines were as follows. (1) The level of micropinocytosis of Noda was significantly less than that of conventional human Mphi cell lines, and the formation of a clathrin-coated pit was not observed in Noda. (2) The level of macropinocytosis of Noda was also smaller than that of conventional Mphi cells indicating that the active membrane ruffling of Noda induces rapid recycling. (3) Phagocytosis of opsonized sheep red blood cells (SRBC) was performed more efficiently in Noda than in other Mphi cell lines. Collectively, these data suggest that in human bone marrow cells, we can identify a unique DC subtype, B/DC line, which develops through a lymphoid DC differentiation pathway, and DC in this lineage plays an important role in the host immune response because of its effective uptake of a variety of size of antigens by using the skillful membrane ruffling and surface receptors PMID- 11380695 TI - Partial tolerance of subcutaneously transplanted xenogeneic tumour cell graft by Fas-mediated immunosuppression. AB - Certain anti-Fas antibodies, such as RMF2, induce apoptosis of Fas-expressing cells. We applied the Fas/anti-Fas system to induce killing of Fas-expressing immunocytes with resultant immunosuppression. W7TM-1 tumour cells, a rat T-cell line, were inoculated subcutaneously in BALB/c mice and tumour growth was monitored in untreated mice and in mice treated with RMF2. Prior to treatment with RMF2, we examined the expression of Fas in isolated splenocytes and in tumour-infiltrating lymphocytes by flow cytometry and immunohistochemistry, respectively. There was a remarkable increase in Fas-positive lymphocytes, including natural killer (NK) cells, among splenocytes at day 5 after tumour cell inoculation. The number of Fas-positive infiltrating lymphocytes also increased markedly, from day 5 to day 10. We then examined whether RMF2 could induce apoptosis of Fas-positive activated lymphocytes isolated from the spleen at day 5 in vitro. Terminal deoxy (d) -UTP nick end labelling (TUNEL) and Annexin V staining methods showed apoptosis of isolated cells when incubated with RMF2, and typical apoptotic features were confirmed by 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) staining. Furthermore, suppression of cellular and humoral immunity was noted in RMF2-treated mice by mixed lymphocyte reaction and assay of serum levels of immunoglobulin G, respectively. Finally, treatment of animals with RMF2 daily from day 5 to day 9 could maintain the tumour size, while the tumour mass began to diminish in untreated mice immediately after reaching a maximum size. We confirmed the enhancing effects of long-term treatment with RMF2, through the induction of immunosuppression, on the growth of unvascularized xenogeneic tumour cell grafts. PMID- 11380696 TI - Mechanism and therapeutic potential of DNA-based immunization against the envelope proteins of hepatitis B virus in normal and transgenic mice. AB - Two plasmid DNA vectors, pCAGGS(S) encoding the genes of the major envelope protein of hepatitis B virus (HBV), and pCAGGS(S + preS2) encoding the genes of the middle envelope protein were used to study the mechanism and therapeutic potential of DNA-based immunization. Injection of these plasmids into the regenerating bilateral tibialis anterior muscle (TA) of normal C57BL/6 mice induced hepatitis B surface antigen (HBsAg)-specific humoral and cellular immune responses. Seventy-two hours after injection of pCAGGS(S), infiltrating cells including antigen-presenting dendritic cells (DC) were localized around the injection site and HBsAg was expressed by both muscle cells and infiltrating cells. Spleen DC from the mice were exposed to HBsAg for up to 32 weeks after a single injection of pCAGGS(S), because these DC induced the proliferation of HBsAg-specific memory lymphocytes in culture without exogenous HBsAg. A single injection of pCAGGS(S) or pCAGGS(S + preS2) resulted in the clearance of HBsAg in 28 out of 30 HBV-transgenic (Tg) mice. In contrast, more than 7 monthly injections of an HBsAg-based vaccine were required for the clearance of HBsAg in 6 out of 29 HBV-Tg mice. Infiltrating DC at the DNA vaccine injection site may have a role in initiating HBsAg-specific immune response, whereas the persistence of HBsAg exposed spleen DC may contribute to long-lasting immunity. This study also suggested that DNA-based vaccines may be a potent tool for treating chronic HBV carriers. PMID- 11380697 TI - Zymosan enhances the immune response to DNA vaccine for human immunodeficiency virus type-1 through the activation of complement system. AB - In the present study, the adjuvant effect of zymosan on human immunodeficiency virus type-1 (HIV-1)-specific DNA vaccine and the mechanism of this enhancement were studied in a murine model. We coinoculated zymosan with our candidate HIV-1 specific DNA vaccine (pCMV160IIIB) into skeletal muscles of BALB/c mice. Higher levels of both humoral immune response and HIV-specific delayed-type hypersensitivity (DTH) response were observed when zymosan was coinoculated with pCMV160IIIB compared with that obtained using pCMV160IIIB alone. HIV-specific cytotoxic T lymphocyte (CTL) activity was also enhanced. This enhancing activity was suppressed when coinoculated to the fifth complement (C5)-deficient DDD and AKR mice. The enhanced activity was also suppressed when anti-C3 antibody was inoculated to mice intramuscularly. There was significant induction of immunoglobulin G2a (IgG2a) and interferon-gamma (IFN-gamma) in pCMV160IIIB vaccine with zymosan. These results suggest that zymosan-mediated DNA vaccination enhances helper T cell (Th) 1-mediated immunity. The effect is suggested to be based on the consequences of its recruitment and activation of macrophages, dendritic cells or antigen-presenting cells (APC) through complement activation, especially through the alternative pathway. Taken together, these results suggest that zymosan can be an effective immunological adjuvant in DNA vaccination against HIV-1. PMID- 11380698 TI - Protection of turkeys against Chlamydophila psittaci challenge by parenteral and mucosal inoculations and the effect of turkey interferon-gamma on genetic immunization. AB - Plasmid DNA (pcDNA1::MOMP A) expressing the major outer membrane protein of an avian Chlamydophila psittaci serovar A strain was tested for its ability to induce protective immunity against challenge with the same C. psittaci serovar. A combined parenteral (intramuscular injection) and mucosal route (DNA drops administered to the nares) of DNA inoculation was compared to three other, different routes of administration (intramuscular inoculation, DNA drops administered to the nares and aerosol immunization). In addition, the effect of turkey interferon gamma (tIFN-gamma) on intramuscular immunization was evaluated by co-expressing pCIneo::tIFN-gamma. A significant level of protection was observed in turkeys immunized via the combined parenteral/mucosal route, the intramuscular route or by aerosol. Severe clinical signs and lesions were observed in the non-vaccinated control groups, in 80% of turkeys inoculated with a mixture of pcDNA1::MOMP A and pCIneo::tIFN-gamma, and in 60% of turkeys vaccinated with DNA drops administered to the nares. The use of MOMP-based DNA vaccination as a means of preventing severe clinical signs and lesions in a turkey model of C. psittaci infection was demonstrated, as was down-regulation of the immune response by co-expression of tIFN-gamma. PMID- 11380699 TI - Mycobacterium avium infection in CD14-deficient mice fails to substantiate a significant role for CD14 in antimycobacterial protection or granulomatous inflammation. AB - CD14 is a pattern-recognition receptor implicated in the inflammatory response to microbial components such as lipopolysaccharide, peptidoglycan and lipoarabinomannan. In this work, we made use of CD14-deficient (CD14-/-) mice to evaluate the relative importance of CD14 in response to infection with viable, intact cells of Mycobacterium avium in vitro and in vivo. Following co-incubation of either bone marrow-derived macrophages (Mphi) or thioglycollate-elicited peritoneal Mphi from CD14-/- mice with viable M. avium, tumour necrosis factor (TNF) production was significantly reduced and delayed compared to TNF secretion by infected CD14+/+ Mphi. However, following intravenous infection with a M. avium strain of either high virulence (TMC724) or intermediate virulence (SE01), there was no difference in the bacterial loads of lungs, livers or spleens at 3, 5 and 8 weeks postinfection in CD14-/- mice when compared with syngeneic CD14+/+ mice. At these time-points, TNF and interferon-gamma (IFN-gamma) mRNA expression in the liver was similar in infected CD14+/+ and CD14-/- mice, and granuloma formation and expression of inducible nitric oxide synthase within granuloma Mphi was the same in both mouse groups. In conclusion, although the absence of CD14 results in significantly reduced and delayed TNF production in response to stimulation with M. avium in vitro, there is no evidence that CD14 plays a significant role in either the antibacterial defence or the chronic granulomatous reaction to M. avium infection in vivo. PMID- 11380701 TI - cAMP response element modulator (CREM): an essential factor for spermatogenesis in primates? AB - CREM is a cAMP-related transcription factor and alternate promotor usage and splicing generate repressor and activator transcripts of CREM within the testis. CREM activators are highly expressed in post-meiotic haploid germ cells and are essential for spermatid maturation in the mouse model as revealed by gene targeting studies. Analysis of testicular CREM expression in rodent and monkey species, and in men yielded a highly comparable pattern thus suggesting that CREM is of general importance for spermatid development in the mammalian testis. Also, many CREM target genes have been identified in haploid germ cells. Studies in men with spermatogenic disturbance and spermatid maturation arrest demonstrated abnormal CREM expression and altered splicing events. Collectively, the data strongly argue for an essential role of CREM during spermatid maturation in primates. PMID- 11380700 TI - Fas-FasL interaction modulates nitric oxide production in Trypanosoma cruzi infected mice. AB - During acute Trypanosoma cruzi infection in mice, many leucocytes undergo apoptosis. Although apoptosis has been ascribed to increased levels of nitric oxide (NO) and Fas-FasL interaction, the importance of this phenomenon in modulating the host response against T. cruzi is unknown. Herein, the role of NO- and Fas-FasL-induced apoptosis in modulating the immune response to T. cruzi was evaluated using mice deficient in Fas expression (MRL/MpJ-Fas lpr) and inducible nitric oxide synthase (iNOS) knockout mice (iNOS-/-). The results showed that besides decreasing apoptosis induction after infection, impairment of the Fas FasL interaction resulted in decreased NO production, as a consequence of enhanced T helper 2 (Th2) cytokine production. Differently, blockage of NO induced apoptosis resulted in uncontrolled cytokine production, rather than a biased Th2 cytokine pattern. Together, these results suggested that Fas and FasL induced apoptosis could be implied in modulation of the immune response against T. cruzi by interfering with cytokine and NO production during the acute phase of the infection. PMID- 11380702 TI - Role of genitourinary inflammation in infertility: synergistic effect of lipopolysaccharide and interferon-gamma on human spermatozoa. AB - Pro-inflammatory cytokines are elevated in the semen of patients with genitourinary inflammation (GUI). Whether this increase in cytokines in GUI patients plays any critical role in male factor infertility is not clear. The present study investigated the in vitro effects of two important pro-inflammatory cytokines, lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma), on sperm motility, viability, membrane integrity and motion parameters. Washed spermatozoa from healthy donors were incubated with LPS (0.1 mg/mL) or IFN-gamma (0.1 mg/mL) alone or in combination. Sperm motility, viability, membrane integrity and computer-assisted motion were evaluated at various time intervals (0, 30, 60 and 180 min) after treatment. Sperm membrane integrity was analysed using the hypo osmotic swelling test (HOST). LPS and IFN-gamma individually did not alter sperm viability or motility, but their combination showed a significant time-dependent decrease (p < 0.05) in sperm motility, viability and membrane integrity. Sperm motion parameters (straight-line velocity, curvilinear velocity, mean linearity, or amplitude of lateral head displacement) were not affected by LPS or IFN-gamma at the concentrations used in this study. These data suggest that the combination of LPS and IFN-gamma is detrimental to human spermatozoa and may contribute to male factor infertility in patients with chronic GUI. PMID- 11380703 TI - Embryological consideration of drainage of the left testicular vein into the ipsilateral renal vein: analysis of cases of a double inferior vena cava. AB - The right gonadal vein (GV=testicular vein in men, ovarian vein in women) usually drains into the inferior vena cava (IVC) while the left gonadal vein drains into the left renal vein (RV). This anatomical difference induces relatively weak haemodynamics in the left testicular vein (TV) and is considered to be a cause of a left varicocele. In textbooks on embryology, it has been documented that bilateral supracardinal veins (=origin of right and left IVC) and the subcardinal sinus (=origin of RVs and GVs) symmetrically develop during early embryogenesis. However, persistence and regression of the right and left supracardinal veins, respectively, results in drainage of the left GV into the ipsilateral RV. A double IVC (DIVC) commonly originates from a failure of disappearance of the left supracardinal vein. Although there have been a considerable number of case reports on DIVC, little attention has been paid to the anatomy of the left GV in such cases. We report here an autopsy case, a 72-year-old Japanese man, with a DIVC. This case belongs to type BC of McClure and Butler's classification. In this case, it was observed that the right TV drained into the confluence of the right IVC with the ipsilateral RV, while the left TV drained into the left RV in spite of the presence of the left IVC. This case indicates that the embryonic anastomosis point between the subcardinal sinus and the supracardinal vein on the left side is different from that on the right side. Statistical analysis of many case reports of DIVC also suggests that the bilateral supracardinal veins tend to asymmetrically anastomose with the subcardinal sinus during embryogenesis. These data imply that drainage of the left GV into the ipsilateral RV leads to regression of the left supracardinal vein but also to asymmetrical anastomosis between the supracardinal veins and the subcardinal sinus. PMID- 11380704 TI - Lectin affinity of the seminiferous epithelium in healthy and cryptorchid post pubertal boars. AB - The present study describes the sugar content of the seminiferous epithelium, using lectin histochemistry, in healthy boars and in boars with unilateral and bilateral abdominal cryptorchidism. In healthy boars the apical cytoplasm of Sertoli cells exhibited abundant glucosyl (Con A and WGA lectins), galactosyl (HPA, DBA, SBA and PNA lectins), and fucosyl (AAA lectin) residues. Spermatogonia and spermatocytes contained abundant glucosyl (Con A and WGA lectins) and fucosyl (AAA lectin) residues. In spermatids, galactosyl (SBA and PNA lectins) and glucosyl (Con A and WGA lectins) residues increased progressively throughout spermiogenesis, and fucosyl (AAA lectin) residues decreased. As compared with healthy boars, the scrotal testis of unilateral cryptorchid boars showed decreased amounts of fucosyl (AAA lectin) and galactosyl (HPA and DBA lectins) residues on the Sertoli cell apical cytoplasm; spermatocytes exhibited higher content of glucosyl (Con A lectin) residues and spermatids showed altered nature of glucosyl (Con A and WGA lectins) and galactosyl (SBA and PNA lectins) complexes. In abdominal testes of unilateral and bilateral cryptorchid boars, immature Sertoli cells and spermatogonia showed decreased fucosyl (AAA lectin), and increased glucosyl (Con A and WGA lectins) and galactosyl (SBA and PNA lectins) contents. These results suggest that the seminiferous epithelium of healthy boars has polarized activity with the apical compartment implicated in germ cell-Sertoli cell adhesion and interaction, in transport of ions, substrates and fluids, and in acrosomal differentiation. In scrotal testes, unilateral abdominal cryptorchidism could lead to defective germ cell-Sertoli cell adhesion, impaired acrosomal differentiation and increased ionic transport in the apical compartment of the seminiferous epithelium. Unilateral and bilateral cryptorchidism could induce increased ionic transport and membrane permeability in the seminiferous epithelium of abdominal testes. PMID- 11380705 TI - Rabbit sex hormone-binding globulin: expression in the liver and testis during postnatal development and structural characterization by truncated proteins. AB - Although sex hormone binding globulin (SHBG) is found in the blood plasma of adult humans and rabbits and the gene is expressed in their livers, it is not detected in the plasma of adult rodents nor is it expressed in adult rodent livers. Thus the rabbit represents a good model to study the metabolism and function of SHBG in the blood. We have used a cloned rabbit SHBG cDNA to detect mRNA expression in rabbits during the postnatal period, and to construct truncated SHBG proteins for structure/function analysis. The SHBG mRNA appeared in the testis as early as 3 days after birth. The level increased gradually in abundance throughout postnatal development, and attained a maximum at 12 weeks of age when the gonads were fully matured. In contrast, SHBG mRNA in the livers of male and female animals increased to a maximum by 4 weeks of age, and were maintained at this level until 12 weeks before subsiding to the initial levels. The increase and decrease in SHBG mRNA levels in the liver were accompanied by similar changes in serum SHBG. This suggests that SHBG in the blood circulation comes from the liver and this might also provide a source of SHBG for the male reproductive tract before formation of the blood-testis barrier. To elucidate the minimal sequence of rabbit SHBG responsible for steroid-binding, a panel of 13 truncated SHBG proteins was constructed, expressed in Escherichia coli, and biochemically purified for study. It was shown that the complete protein sequence of rabbit SHBG was important for maintaining a stable steroid-protein complex. Unlike human SHBG for which a truncated protein of the first 206 residues of the 373 amino acid protein can still bind steroid, removal of 43 or more residues from the C-terminus of rabbit SHBG completely abolished steroid-binding. PMID- 11380706 TI - High incidence of single nucleotide substitutions in the mitochondrial genome is associated with poor semen parameters in men. AB - Single nucleotide polymorphisms (SNPs) in 7000 bp of the mitochondrial genome, encompassing 15 coding regions from COI to ND5, were characterized by single strand polymorphism analysis and confirmed by DNA sequencing. About 2.4% of normozoospermic men and 8.4% of men with poor semen quality had at least one nucleotide substitution. Most of the substitutions occurred in the third codon and did not change the amino acid. Hydrophobicity plots of the proteins with changes in an amino acid as a result of a nucleotide substitution suggested that they did not affect the function of the protein. The two most common substitutions at nucleotide (nt) 9055 and 11719 had significantly higher frequencies in men with reduced sperm motility. Eleven percent of the men with poor semen parameters and 1.3% of normozoospermic men had a 9055 substitution, 12% of the men with poor semen parameters had a substitution at nt 11719, but none of the normozoospermic men had this substitution. All the patients with these substitutions had reduced sperm motility and/or low sperm count. These SNPs in the mitochondrial genome were in a homoplasmic state. Thus, we propose that possessing these mitochondrial mutations compromises the semen quality of these men. PMID- 11380707 TI - Novel amino acid substitutional mutation, tyrosine-739-aspartic acid, in the androgen receptor gene in complete androgen insensitivity syndrome. AB - Androgen insensitivity syndrome (AIS) is an X-linked recessive disorder. The molecular mechanism of AIS is reduction or absence of androgen signalling caused by androgen receptor (AR) malfunction or absence. The phenotype of AIS varies from a complete female phenotype (complete AIS, CAIS) to male genitalia with mild hypospadias (partial AIS, PAIS). In the current study, we characterize a novel point mutation in the ligand binding domain of the AR gene in a 50-year-old Japanese CAIS patient. Sequence analysis showed a single point mutation at nucleotide 3359 (Genbank, NM 000044), T to C, in exon E in the AR gene. This mutation led to the conversion of codon 739 tyrosine into aspartic acid in the ligand binding domain. No specific androgen binding was detected in genital fibroblasts isolated from the patient. Transcriptional activating activity of the mutant AR was examined by transient DNA transfection into COS-1 cells. Wild-type AR successfully activated androgen inducible MMTV promoter dose-dependently. In contrast, the mutant AR did not activate MMTV promoter. Thus, we demonstrated the molecular characteristics of the novel point mutation in the ligand binding domain of the AR gene associated with CAIS. This information will provide a further understanding of the structure and function of the AR gene. PMID- 11380708 TI - Nursing education in Ireland: the end of the beginning and the envy of others in Europe. PMID- 11380709 TI - Back to the bedpans: the debates over preregistration nursing education in England. AB - AIMS OF THE PAPER: This paper aims to examine the recent debates in both the general and professional press over the future of preregistration education in England, in the light of the sociological literature on the professions, and the nature of nursing knowledge. The literature on the Project 2000 reforms, although by now well worn, will also be discussed briefly. BACKGROUND/RATIONALE: The bulk of nursing education in England moved into higher education in the 1990s. Unlike other higher education in England, it is not funded by the Department for Education and Employment, but commissioned by the National Health Service (NHS), to which it is expected to be highly responsive. DESIGN/METHODS: This paper is based on a brief literature survey, plus policy documents and media reports. FINDINGS/CONCLUSION: It will be argued that the process of professionalization is not a straightforward one, and has its potential for reversals of fortune, and that nursing occupies a marginal place in higher education in England. PMID- 11380710 TI - Emotion work in midwifery: a review of current knowledge. AB - AIM OF THE PAPER: To review the literature relating to emotional labour in the workplace and identify potential sources of emotion within midwifery work. RATIONALE: There is substantial evidence to indicate that the quality of the relationship between midwife and woman is significant in determining the quality of the childbirth experience for women. Despite this, there is a notable lack of research regarding midwives' experiences of participating in this relationship, and even less regarding the emotional issues involved. METHOD: Literature review of relevant midwifery, nursing and sociological literature. Discussion of the theoretical perspectives provided by sociological and nursing research relating to the management of emotion at work and critical consideration of their application to an analysis of midwifery work. FINDINGS: Although these theoretical perspectives may offer significant insights of relevance to midwifery, there is much more that needs to be uncovered. Midwifery work has the potential for creating high levels of emotion work and current changes in the organization of United Kingdom (UK) maternity care may further increase this. CONCLUSION: It is essential that midwives develop their understanding of emotion at work in order to improve their own working lives, and to meet the needs of childbearing women and their families. More research is needed in this field to develop a body of knowledge to inform midwifery education and practice. PMID- 11380711 TI - The relationship between attendance at birth and maternal mortality rates: an exploration of United Nations' data sets including the ratios of physicians and nurses to population, GNP per capita and female literacy. AB - The relationship between attendance at birth and maternal mortality rates: an exploration of United Nations' data sets including the ratios of physicians and nurses to population, GNP per capita and female literacy. BACKGROUND: This is the third and final paper drawing on data taken from United Nations (UN) data sets. The first paper examined the global distribution of health professionals (as measured by ratios of physicians and nurses to population), and its relationship to gross national product per capita (GNP) (Wharrad & Robinson 1999). The second paper explored the relationships between the global distribution of physicians and nurses, GNP, female literacy and the health outcome indicators of infant and under five mortality rates (IMR and u5MR) (Robinson & Wharrad 2000). In the present paper, the global distribution of health professionals is explored in relation to maternal mortality rates (MMRs). The proportion of births attended by medical and nonmedical staff defined as "attendance at birth by trained personnel" (physicians, nurses, midwives or primary health care workers trained in midwifery skills), is included as an additional independent variable in the regression analyses, together with the ratio of physicians and nurses to population, female literacy and GNP. AIM: To extend our earlier analyses by considering the relationships between the global distribution of health professionals (ratios of physicians and nurses to population, and the proportion of births attended by trained health personnel), GNP, female literacy and MMR. 0.05). The findings were not, however, deemed clinically significant in that no change in care or treatment resulted from differences in temperature recordings. CONCLUSIONS: Whilst the findings cannot be applied to sick preterm neonate, it was concluded that tympanic membrane temperature recordings in healthy preterm neonates are safe, accurate, easy, and comfortable for the baby, and appropriate with this client group provided staff are trained in the technique. PMID- 11380714 TI - No more tears: a randomized controlled double-blind trial of Amethocaine gel vs. placebo in the management of procedural pain in neonates. AB - AIM: During a comparative double-blind study, the efficacy of Amethocaine local anaesthetic gel was compared with that of a placebo gel to establish their effect on the reduction of behavioural and physiological responses to pain in neonates. METHOD: Forty infants were included in the trial, 20 being randomly assigned to each group. After topical application of either Amethocaine or the placebo for 30 minutes, cannulation was performed using a 24 gauge cannula. Assessment of pain was made with the aid of four point assessment tool incorporating facial expression, cry and heart rate changes and ease of cannulation. Additional baseline data were obtained including sex, gestation, weight and previous experience of cannulation, and analysed using Statistical Package for Social Sciences (SPSS). RESULTS: Good analgesic effects were demonstrated in the Amethocaine group, as there was a high level of significance (P < 0.01) in the difference in pain scores between the two groups using a Mann-Whitney U-test. There was no correlation between gestation or weight and the degree of pain experienced during painful procedures. No significant relationship was found between cannulation and the degree of pain experienced. CONCLUSION: It can, therefore, be concluded that Amethocaine gel is an effective local anaesthetic in the management of skin puncturing procedural pain in neonates, and that all infants regardless of maturity, weight or previous experiences probably experience a similar degree of pain and therefore warrant effective and appropriate analgesic intervention. PMID- 11380715 TI - Nonpharmacological methods in relieving children's postoperative pain: a survey on hospital nurses in Finland. AB - AIM OF THE STUDY: The aim of this study was to describe nurses' use of selected nonpharmacological methods in relieving 8-12-year-old children's postoperative pain in hospital. METHODS: The convenience sample consisted of 162 nurses working on the paediatric surgical wards in the five Finnish university hospitals. An extensive questionnaire, including a five-point Likert-scale, on the nurses' use of selected nonpharmacological methods and demographic data was used as a method of data collection. The response rate was 99%. Descriptive statistics as well as nonparametric Kruskall-Wallis ANOVA and the chi-squared test were used as statistical methods. RESULTS: The study indicates that emotional support, helping with daily activities and creating a comfortable environment were reported to be used routinely, whereas the cognitive-behavioural and physical methods included some less frequently used and less well known strategies. The results also show that attributes, such as the nurses' age, education, and work experience, the number of children the nurses had, the nurses' experiences of hospitalization of their children as well as the hospital and the place of work, were significantly related to the use of some nonpharmacological methods. CONCLUSIONS: The nurses used versatile nonpharmacological methods in children's postoperative pain relief, although some defects could be observed. More research is needed on the methods used by nurses to relieve children's pain in different patient groups and the factors which hinder or promote nurses' use of pain alleviation methods in the clinical practice. PMID- 11380716 TI - Experiences of well spouses after lung transplantation. AB - AIM OF THE STUDY: To explore what life is like for well spouses after their partners' lung transplantation? BACKGROUND: The numbers of people having a lung transplant in the United States of America (USA), as well as surviving beyond 1 and 5 years, has increased steadily over the last decade. This trend is expected to continue. Few have examined the transplant experience from the view of well spouses who are the most typical patient-support person. Researchers have shown that spousal adjustment is closely associated with patient adjustment and recovery. DESIGN/METHODS: Using written surveys and taped telephone interviews in 1998, a convenience sample of 12 well spouses shared details of their lives after their partners' lung transplant. Spouses from eight states in the USA responded to an invitation to participate which was posted on an internet listserv. Ages ranged from 40 to 66 years and the posttransplant time ranged from 4 to 60 months. Spouses completed a demographic form, the Family Inventory of Life Events (FILE), and the Center for Epidemiologic Studies Depression Scale (CES-D) prior to interview. Triangulation of methods augmented and corroborated data. Interviews stopped when saturation was reached. Data collection and analyses were carried out simultaneously using a constant comparative method. RESULTS: The core theme was the "roller coaster ride" characterized by a series of ups and downs. Other major themes were coping, giving medications, knowing the donor, making comparisons, togetherness, and caring for the well spouse. Five identified stages were: the transplant event, cocooning, normalizing, branching-out and settling down. CONCLUSIONS: This study supports nurses using a family centred focus. Nurses in all practice areas can apply information gained from this study to help spouses anticipate chaos at each stage and develop effective coping strategies. PMID- 11380717 TI - Forced externalization of control in people with diabetes: a qualitative exploratory study. AB - AIM: The purpose of this study was to explore and describe the patients' perceptions of the Diabetic Disease State and its complications using an ethnographic method. BACKGROUND: Diabetes mellitus is an extensively researched and studied disease, however, relatively little attention has been given to the lived experience of chronic illness. METHODS: A purposive sample of 18 people with diabetes was derived from a diabetes register database. Data were collected by semistructured interviews, which were tape-recorded and transcribed. The transcripts were analysed together with the researchers' observational notes, using open coding procedures to identify main categories. Discussion of the results is made in the theoretical context of the locus of control (LOC) framework. FINDINGS: Three main categories emerged from the data: information knowledge of illness; the psychological burden of diabetes; rationalizing. A further subcategory, reality avoidance, was identified. The findings showed that these people with diabetes experience and describe complex psychological effects and coping mechanisms in having a chronic illness. CONCLUSIONS: The findings also suggested that participants in this study might have felt that the control of their chronic condition had been externalized to the health care professionals responsible for their care. PMID- 11380718 TI - Power and self-perceived health in blind diabetic and nondiabetic individuals. AB - AIMS: The aims of this study were: (1) to explore perceptions of power in blind individuals and relate presence or absence of power to self-perceived health and (2) to compare self-perceived health in blind individuals with that of the general population. BACKGROUND: The theoretical framework of this study was Barrett's Power theory, which is based on The Rogerian nursing theory. Power is defined as being aware of what one is choosing to do, feeling free to doing it, and do it intentionally. METHODS: Thirty-nine blind subjects at three adjacent ophthalmology centres agreed to participate in the study. Of those 23 had become blind because of late complications of diabetes. Power was explored during semi structured interviews and self-perceived health was measured with the Swedish health-related quality of life questionnaire. Data on socio-economic, rehabilitative and diabetes-related variables were also collected. FINDINGS: Power was experienced by 19 of the 39 respondents and was more frequently found in nondiabetic subjects than in diabetic subjects. Those experiencing power reported a better emotional and general health compared with individuals lacking power. The perception of having power was not significantly related to any other of the studied variables. When compared with age- and gender-matched controls from the general population, nondiabetic blind subjects scored higher in positive feelings and lower in physical functioning. In contrast diabetic subjects experienced poorer general health, less satisfaction with physical health and more negative feelings, but they reported that they did not experience limitation as a result of these emotions. CONCLUSION: One way of improving health in diabetic blind individuals could be to increase the subject's perception of power. PMID- 11380719 TI - Patients with congestive heart failure and their conceptions of their sleep situation. AB - AIM: To describe, from a nursing perspective, how patients with CHF conceived their sleep situation. BACKGROUND: Sleep disturbances are very common in patients with congestive heart failure (CHF). Polysomnographic studies have shown that the total duration of sleep is shorter and the sleep structure disturbed, with frequent arousals and sleep stage changes. METHODS: A qualitative descriptive design inspired by a phenomenographic approach was employed. Conceptions were collected through interviews with 20 strategically chosen CHF patients. FINDINGS: The findings showed that the patients' sleep was affected by their daily activities, the disease itself and cardiac symptoms. The sleep disturbances gave effects such as fatigue, listlessness, loss of concentration and loss of temper. These effects led to a need for daytime sleep, seclusion, counselling and information. Patients handled their sleep disturbances through coping mechanisms related to developed patterns of daily life and through support from their psychosocial environment. CONCLUSIONS: Through an increased awareness of the causes of sleep disturbances in CHF patients, nurses can more effectively meet their caring needs and reduce the psychological stressors that patients develop. Information and education, both to patients and the next of kin, about the disease and the sleep situation, especially good sleeping habits, can help patients to better cope with sleep disturbances. PMID- 11380720 TI - Mutual relating: developing interpersonal relationships in the community. AB - AIM OF THE PAPER: This paper, which is part of a larger study that examined how community mental health nurses promoted wellness with young adult who were experiencing an early episode of psychotic illness, aims to explicate how nurses develop interpersonal relationships with these consumers. RATIONALE: Research into the development of interpersonal skills is important because they are an essential foundation to the promotion of wellness in psychotic illness. DESIGN/METHODS: The study used Strauss and Corbin's approach to grounded theory methodology. Concurrent data collection and analysis was undertaken incorporating interviews and observations. The study took place in the community, in rural and regional New South Wales, Australia, and involved consumers, significant others, and community mental health nurses. RESULTS: The findings show that several strategies are associated with mutual relating. Attempting to understand necessitates nurses trying to see it from insiders' points of view, while acknowledging that they can never completely understand their illness experience. Being friendly indicates that the qualities of a friendly relationship should be implicit in a professional friendship. Tuning in involves taking account of and adjusting to consumers' ways of dressing, communicating, and the types of approaches that consumers prefer. Revealing oneself is a process that helps put both consumers and nurses at ease, and helps dispel the perception consumers take part in a one-sided relationship. Being there for them, is a particular type of obligation that entails nurses being accountable for the care they provide, being a spokesperson for consumers, if required, and avoiding collusion with others. Related to avoiding collusion, is the issue of maintaining confidentiality, particularly within the context of interacting with significant others. CONCLUSION: The results of the study have implications for the way nurses develop and maintain interpersonal relationships with consumers, and this is affected by nurses' personal and professional attributes. PMID- 11380721 TI - Finnish occupational health nurses' work and expertise: the clients' perspective. AB - AIMS: The aims of this study were to describe Finnish occupational health nurses' functions, characteristics, prerequisites, consequences, changes, development areas and expertise from the point of view of clients. METHODS: The background literature of this study is based on public health nursing models, Finnish social and health report, arguments of special education for occupational health nurses, and earlier studies concerning occupational health nurses' work. The data were collected from volunteer clients (n=26) by interviews. RESULTS: According to the qualitative content analysis, occupational health nurses' activities include health promotion and secondary health care among workers and at workplaces. The main work characteristics are holism, client-orientation, interaction and co operation. Occupational health nurses need an extensive knowledge base and practical skills, client-orientation, courteous behaviour and a healthy and clean appearance. The outcomes of their work for clients are better health, healthier life habits and healthier working conditions. Nowadays, nurses are more client orientated than 20 years ago. They are expected to develop their practical and interaction skills and expand their knowledge base. The expertise of occupational health nurses consists of an extensive knowledge base with practical skills, working experience and confidence, and it appeared when advising clients and answering their questions. CONCLUSIONS: It is important to arrange continuing education for occupational health nurses to ensure that they are always up to date in order to be able to respond to specific clients' needs. This study provides a foundation for further investigations into, for example, occupational health nurses' work from the point of view of employers, students of occupational health nursing and other occupational health experts and co-operative partners. PMID- 11380722 TI - The utility of self-care theory as a theoretical basis for self-neglect. AB - AIM: This paper sets out to explore the utility of self-care theory in understanding self-neglect. Further theoretical development of both self-care and self-neglect theory and attending core concepts is an important objective. BACKGROUND: The notions of self-neglect and self-care are frequently linked in the literature. The relationship between self-neglect and self-care is not clear and the strengths and limitations in using self-care theory to facilitate a greater understanding of self-neglect will be addressed. Specifically the issues of self-care agency, self-care requisites, objectivity, class and culture, and lifestyle choice will be critically evaluated in the context of self-neglect theory. CONCLUSION: Self-care theory has a useful role to play in furthering our understanding of self-neglect. Self-care theory is able to explain some aspects of self-neglect but not others, although this may be a reflection of the relatively underdeveloped state of self-care theories or alternatively may reflect a more fundamental limitation in our ability to fully explain human behaviour. PMID- 11380723 TI - Response to: Lauder W.(2001)The utility of self-care theory as a theoretical basis for self-neglect. Journal of Advanced Nursing 34(4), 545-551. PMID- 11380724 TI - Development of a theoretical framework describing relatives' involvement in palliative care. AB - BACKGROUND: The present study is based on four earlier studies in which the authors classified the relative's involvement in palliative care into different categories and described the involvement as "involvement in the light" or "involvement in the dark". AIM: The aim of the study was to develop a theoretical framework concerning the involvement of relatives based on an in-depth analysis of the results of the four earlier studies. METHOD: Walker & Avant's (1995) strategies for theory construction were used for development of the framework. A number of different concepts, assumptions and statements about relatives' involvement were penetrated in an in-depth analysis. RESULTS: From the concepts two theoretical "blocks" of the relatives' involvement were developed and these constitute the foundation for the framework. One is based on concrete descriptions of the concepts "to know", "to be" and "to do". The other describes how the concepts of "involvement in the light" and "involvement in the dark" differ. Factors that promoted involvement in the light were professional care based on humanistic values, a stronger sense of coherence on the part of relatives, an appropriate illness trajectory, and other available resources. The opposite was the case for those who were involved in the dark. Five assumptions successively developed which together form the theoretical framework. CONCLUSIONS: An important conclusion that can be of importance in palliative care is that the manner in which the staff act toward the patient and relatives influence relatives' possibilities for involvement, patients' possibilities for an appropriate death, and the possibilities the staff have to give good care. PMID- 11380725 TI - Myth of empowerment in chronic illness. AB - AIMS: This article presents several findings of a study, conducted between 1996 and 1998, to investigate self-care decision making in diabetes. RATIONALE: The underlying assumption of many practitioners is that an invitation to people with chronic illness to participate as equal partners is sufficient to guarantee their empowerment. DESIGN: Using grounded theory, the research examined self-care decision making using a convenience sample of 22 Canadian adults with longstanding type 1 diabetes nominated as expert self-care managers. Participants audiotaped their decision making as it occurred for 3 weeks over the course of one calendar year. These audio-recordings were followed by an interview to clarify participants' decision making and factors that affected their decisions. FINDINGS: Participants identified several covert and subtle ways that practitioners contradict their stated goal of empowerment in their interactions with diabetics. Participants revealed that despite their intention to foster participatory decision making, practitioners frequently discount the experiential knowledge of diabetes over time and do not provide the resources necessary to make informed decisions. CONCLUSION: The article concludes with a discussion of the implications of the findings for practice. PMID- 11380726 TI - Between joy and sorrow: being a parent of a child with developmental disability. AB - AIM: This study explored the experiences of parents who have children with significant developmental disability. BACKGROUND: Prevailing societal and professional assumptions of parental crisis and maladjustment in response to the 'tragedy' of having a disabled child did not accord with the authors' practice experience. Whilst parents confronted numerous difficulties, most of them appeared to manage with optimism and remarkable resourcefulness. RESEARCH DESIGN: The study, using an interpretive methodology informed by phenomenology, intensively explored the experiences of six parents of children with significant developmental disability. FINDINGS: Although they experienced much anguish and sorrow, the parents also spoke of hope, love, strength and joy. Interpretation of the parents' experiences revealed the themes of 'joy and sorrow', 'hope and no hope' and 'defiance and despair', mediated by 'the tensions'. CONCLUSIONS: This phenomenological interpretation provides insight and understanding into the parents' experiences and has implications for practice, education and research in nursing. PMID- 11380727 TI - Negotiating lay and professional roles in the care of children with complex health care needs. AB - BACKGROUND: Children with complex health care needs are now being cared for at home as a result of medical advances and government policies emphasizing community-based care. The parents of these children are involved in providing care of a highly technical and intensive nature that in the past would have been the domain of professionals (particularly nurses). AIMS OF THE PAPER: To assess how the transfer of responsibility from professionals to parents was negotiated, the tensions and contradictions that can ensue, and the implications for professional nursing roles and relationships with parents. DESIGN: Using a Grounded Theory methodology, in-depth interviews were conducted with 23 mothers, 10 fathers and 44 professionals to gain insight into the experience of caring for children and supporting families in the community. FINDINGS: From the parents' perspective, their initial assumption of responsibility for the care of their child was not subject to negotiation with professionals. Prior to discharge, parents' feelings of obligations, their strong desire for their child to come home, and the absence of alternatives to parental care in the community, were the key motivating factors in their acceptance of responsibility for care-giving from professionals. The professionals participating in the study had concerns over whether this group of parents was given a choice in accepting responsibility and the degree of choice they could exercise in the face of professional power. However, it was following the initial discharge, as parents gained experience in caring for their child and in interacting with professionals, that role negotiation appeared to occur. CONCLUSION: This study supports other research that has found that professionals' expectations of parental involvement in the care of sick children role can act as a barrier to negotiation of roles. In this study, parental choice was also constrained initially by parents' feelings of obligation and by the lack of community services. Nurses are ideally placed to play the central role not only in ensuring that role negotiation and discussion actually occurs in practice, but also by asserting the need for appropriate community support services for families. Being on home territory, and in possession of expertise in care-giving and in managing encounters with professionals, provided parents with a sense of control with which to enter negotiations with professionals. It is important that changes in the balance of power does not lead to the development of parent-professional relationships that are characterized by conflict rather than partnership. PMID- 11380728 TI - Dilemmas in the provision of own-home respite support for parents of young children with complex health care needs: evidence from an evaluation. AB - Dilemmas in the provision of own-home respite support for parents of young children with complex health care needs: evidence from an evaluation Aims. The aim of this article is to present some of the results of a qualitative evaluation of a United Kingdom (UK) nurse-led, home-based, respite service for the families of children under the age of five with complex health care needs. Background. Advances in neonatal medical and nursing care have contributed to a growth in the number of children who survive low birth weight, birth trauma, and various congenital anomalies. Many of these children are likely to have long-term care needs which will require innovative nursing responses. Of particular importance is the need for parental respite, given the added demands of caring for very ill children at home. Methods. A parent-centred, follow-up evaluation, using in-depth qualitative interviews with parents in 18 families consecutively referred to the Children's Outreach Service between April and December 1997. Findings. This evaluation reveals the sometimes mixed reactions of parents to this innovative service, and the equivocal evidence about its role in contributing to family well being. We discuss, from the perspective of parents using the service, some of the dilemmas and problems in the provision of home-based respite support to this client group. In particular, we explore the diverse ways in which families talk about their need for respite support and point to the need for flexibility in this kind of service provision if these support needs are to be met. Conclusion. We conclude by discussing the practice implications of our evaluation for those planning similar services. PMID- 11380730 TI - Whither nursing? Discourses underlying the attribution of master's level performance in nursing. AB - AIM: Drawing upon the selected findings from a multidisciplinary study that sought to explore the meaning of master's level performance in health professional practice, the characteristics which nurse educators attributed to the practice of master's level nursing graduates are analysed to reveal underlying discourses. BACKGROUND: Although master's level programmes for nurses have been available in the United Kingdom (UK) for the past three decades and current heath policy directives link master's level qualifications with senior clinical nursing roles, the contribution that master's level education might make to the future direction of nursing is unclear. RESEARCH DESIGN AND METHODS: In depth interviews were undertaken with a purposive sample of 18 nurse lecturers drawn from eight universities in the UK who were responsible for master's level programmes in nursing. The interview agenda explored participants' perspectives of the characteristics of master's level performance. Drawing upon the methodology of discourse analysis, interview transcripts were interpreted in such a way as to show the implicit discourses underlying the participants' claims regarding their graduate's attributes of professional practice. FINDINGS: The characteristics attributed to master's graduates were categorized under (a) cognitive competencies, (b) practice-related competencies, (c) research orientation and (d) personal dynamism. However, these attributions are not empirical generalizations, developed inductively. Rather, they draw on socially available discourses regarding the future direction of the profession. CONCLUSION: The nurse educators drew on the following socially available discourses: (a) a discourse in which nursing is construed as involving great competence in practice, but without radicality of thought. Associated with this is a pervasive rhetoric of pragmatism; (b) a discourse of interprofessional practice in which nursing has a role of leadership. This is associated with a view of the location and power of nursing within the structure of the National Health Service and (c) notably lacking were discourses of care-giving, and of academic/intellectual aspiration. The implications of these discourses for the future direction of nursing are considered. PMID- 11380729 TI - The effectiveness of parenting programmes facilitated by health visitors. AB - AIM OF THE STUDY: This study was designed to evaluate the effectiveness of parenting programmes that are facilitated by health visitors and offer practical and emotional support to parents in the Down Lisburn Trust, Northern Ireland. METHODS: All parents (n=78) taking part were given a self-administered questionnaire both before and after attending a 'Positive Parenting' programme. The questionnaire contained items gathering demographic information, standardized measures of clinical anxiety/depression, personality states and coping styles and participants' experience of being a parent and what they wanted from the parenting group. Two items adapted from the Parent Stress Index looked at participants' ratings of themselves as parents, and if they enjoyed this role. STATISTICAL ANALYSIS: All statistics were computed using the SPSS for Windows (version 7) statistical package. The parametric paired-samples t-test was used with the ratio data generated by the Hospital Anxiety and Depression Scale (HADS) questionnaire. It is used to test the difference between the means of the pre and postsets of scores for significance. The nonparametric Wilcoxon test was used to find if there was a significant difference between the pre and postprogramme related samples by ranking the means. The Wilcoxon was chosen firstly because the data were ordinal in nature, hence it cannot be assumed that the findings follow the normal curve of distribution and secondly because it cannot be assumed that the data have homogeneity of variance. FINDINGS: At the end of the 8-week parenting programme, findings demonstrated significant statistically reduced levels of clinical anxiety and depression. Parents demonstrated an increase in more positive ratings of personality states such as not shouting at their children and being more calm and energetic at the end of the programme. Adaptive problem focused coping strategies were reported as being used more often subsequent to the programme, although some of the less adaptive emotion focused strategies had also increased in use. However, no change was evident in their ratings of how good a parent they were or if they enjoyed being a parent. CONCLUSIONS: Health visitors have the skills and are in the position to be preventative agents of inadequate parenting, and advocates of positive parenting. Factors relevant to the realm of positive parenting are highlighted as possible avenues for future research. PMID- 11380731 TI - The primary and secondary care interface: the educational needs of nursing staff for the provision of seamless care. AB - AIM: To identify nurses' perceived deficits in the knowledge and skills required to provide effective seamless care, so that appropriate training could be provided. BACKGROUND: A clear understanding of nursing staff roles, skills and resources is paramount to work at the primary/secondary care interface. Nursing staff require an educational model that will provide a clear understanding of how their roles coalesce with other healthcare professionals. There is little evidence that examines the educational needs of nurses related to changing care boundaries. DESIGN/METHODS: The study used methodological triangulation to explore these issues within current practice. Focus groups were used to generate items for inclusion in the questionnaire. Questionnaire design was based on an importance-performance analysis. This procedure has been effective in developing health care marketing strategies. A stratified random sample of nursing staff (n=722) from the participating trusts received the questionnaire, eliciting a response rate of 172 (23.8%). RESULTS: Factor analysis provided a list of seven training categories in order of training need priority: information technology, awareness of roles, communications within seamless care, working across boundaries, professional issues, practice-related issues, delivery of patient/client care issues. There were no differences in nurses' training needs across NHS trusts. However, differences were highlighted for staff located in primary or secondary settings or working across the interface. CONCLUSIONS: Despite there being a vast range of training issues the majority of nurses appear to have a clear idea of their training needs for the provision of seamless care. A training programme required which targets the specific needs of nursing staff working at different positions across the primary/secondary care interface. PMID- 11380732 TI - Reflection as a transforming process: student advanced nurse practitioners' experiences of developing reflective skills as part of an MSc programme. AB - AIM OF THE STUDY: The aim of this study was to explore student advanced nurse practitioners' (ANPs) experiences of reflection. BACKGROUND: The implementation of a National Health Service (NHS) Executive initiative to develop a Master's Course in advanced nursing practice at Wolverhampton University provided the background to the study. The course was designed to enable experienced nurses to take on activities previously considered the province of physicians. The aim was to extend practice within a holistic nursing framework. Reflection was perceived as central to this process. A pre-entry degree level reflective practice module and a reflective component for the level 4 (Master's level) Practice Module were devised to enhance practitioners' reflective skills. RATIONALE: The rationale for the study was that it should explore student ANPs' perceptions of reflection following completion of these modules, to evaluate their experiences and inform curriculum development. DESIGN/METHODS: A qualitative methodology was used. The sample comprised 14 ANP students. Interviews and reflective learning contracts were used to collect the data. Colaizzi's (1978) seven-stage model guided data analysis. FINDINGS: The findings indicated that all but one of the students described the development of their reflective skills positively. Changes are described in students' thinking and behaviour. CONCLUSIONS: The conclusions have implications for ANPs, for patient care, and for others seeking to develop the ANP role. PMID- 11380733 TI - The development of reflective learning in the context of health counselling and health promotion during nurse education. AB - AIMS AND BACKGROUND: The purpose of this follow-up study was to describe the development of reflective learning of 16 student nurses in the context of health counselling and promotion during clinical training of a 3-year nursing education programme in two Finnish polytechnics. The first aim was to analyse the levels of reflectivity in the student nurses' reflections on their videotaped counselling situations. The second aim was to explore the qualitative features of the reflective health counselling learning process in the context of health promotion. The theoretical background of the study was based on a transformative learning theory. METHODS: The data consisted of stimulated recall interviews with 16 student nurses conducted once a year between 1998 and 2000. The data were analysed using categorization and thematic analysis. FINDINGS: The findings of the study showed that half of the students in the research group reached the level of critical consciousness during their nursing education. The others remained at the level of consciousness. Two students who represented thoughtful action without reflection in the first study year were able to reach reflection and even critical reflection later during their education. The meaning schemas of counselling developed and were enriched when the students moved into the higher stages of reflection. There were features of an empowerment approach to health promotion only in the meaning schemas and perspectives of critical reflectors. CONCLUSIONS: These results may indicate that the empowerment approach to health promotion within the modern health promoting role of nurses requires critical reflection. Therefore, effective methods and evaluation tools of reflective learning are needed to support learning from practice via critical reflection. PMID- 11380734 TI - Chronic pain and coping: a proposed role for nurses and nursing models. AB - AIMS: Activity of living based nurse models such as that proposed by Roper, Logan and Tierney are utilized commonly to rationalize, structure and guide practice. This paper proposes that their use, in the context of chronic pain, should be extended to incorporate the assessment of coping and its effect on disability. BACKGROUND: As coping has been found to account for some of the variance in adaptation to chronic pain, generic models of coping and their utility in this context are considered. The need for reliable and valid means to assess coping are of paramount importance as this will facilitate and inform treatment and management of chronic pain. At the present time coping is most generally assessed using self report instruments which primarily focus on the number and type of strategies used to cope rather than their effectiveness in reducing disability and increasing function. FINDINGS: A classification system which categorizes chronic pain patients in terms of how they function in daily life as either adaptive copers, interpersonally distressed or dysfunctional is outlined. The similarities between this and a routine nursing assessment based on an activities of living model are highlighted. CONCLUSIONS: It is proposed that the innovative use of nursing activity of living models could provide the framework for a pain clinic based assessment of coping that has greater utility for professionals and patients alike. PMID- 11380735 TI - Intentional action. AB - AIM: The purpose of this paper is to present a concept analysis of intentional action. BACKGROUND: Intentional action is a relevant concept for nurses because health behaviour involves action by the client of care. In addition, nurses take action when they assist their clients. Intentional action is a type of action distinguished by the presence of specific developmental skills and by intent or purpose. DESIGN: This concept analysis reflects a comprehensive review of the action literature. FINDINGS: This review revealed two views of intentional action: as social behaviour and as self-regulated behaviour influenced by external and internal factors. Based on these findings, the antecedents and consequences of this type of action are identified and the process of action is described. CONCLUSIONS: The concept model illustrates the relationship of intentional action with external and internal factors and demonstrates its continuous nature. Areas for further inquiry include exploration of the effect of factors such as self-efficacy, motivation, self-esteem, managed care and reduced health care resources on intention and subsequent action; as well as the efficacy of nursing interventions designed to influence health behaviour such as health promotion education and treatment counselling. PMID- 11380736 TI - A measure of patient empowerment in hospital environments catering for older people. AB - AIM: The principal aim of this research was to develop a valid and reliable measure of patient empowerment and disempowerment in hospital environments catering for older people. Rationale. It is argued that empowerment is linked to Learned Mastery (Peterson, Maier & Seligman 1993) and increasing patient independence, whilst disempowerment is linked to Learned Helplessness (Seligman 1975) and increasing patient dependence. Consequently, the development of a measure of these concepts would represent a valuable quality assurance tool. METHOD: Measures were developed using the 'act frequency approach' of Buss and Craik (1983) whereby registered nurses (n=38) were asked to nominate empowering and disempowering acts relevant to interactions between staff and older patients. The resulting lists of 98 acts for each disposition were hypothetically judged by older hospitalized people (n=20) as to the extent they would be either 'control giving' (empowering acts) or 'control taking' (disempowering acts) if personally experienced. The twenty highest scoring acts in each category were incorporated into the Patient Empowerment Scale. RESULTS: Using this scale, older hospitalized people (n=102) resident on five different hospital wards were asked to judge how often they had encountered each act over a predetermined period of time. Wards varied in speciality including acute medicine, surgery and elderly care rehabilitation. The ward scoring lowest on the PES (i.e. the least empowering) was the elderly care rehabilitation ward. Moreover, this ward showed an inverse correlation between participant age and exposure to empowering care. Alpha scores ranged from 0.74 to 0.87 for empowerment and 0.65 to 0.87 for disempowerment. CONCLUSION: The PES is offered as a valid and reliable measure of empowerment and disempowerment in hospital settings catering for older people. It enables the identification of environments which place patients at risk of becoming dependent, or alternatively facilitate increasing independence. PMID- 11380737 TI - The role of tacit knowledge in the work context of nursing. AB - AIMS OF THE STUDY: Previous research on the role of tacit knowledge is ambiguous. Some studies show the superiority of expertise, while other studies found experts would not be better than laymen. This paper aims at clarifying the contribution of tacit knowledge to expertise in the domain of nursing. BACKGROUND: Two important concepts for dealing with critical situations are outlined - tacit knowledge and experience-guided working. The framework of tacit knowledge and experience-guided working can contribute to an explanation of the ambiguous results. Tacit knowledge is acquired implicitly in the course of working and is therefore not subject to reflection. For this reason it can contain erroneous or problematic contents. METHODS: A method for the explication of tacit knowledge was developed and a laboratory study with 16 experienced nurses conducted. In the laboratory study the nurses had to deal with a critical nursing situation that was developed in co-operation with nursing experts. The explicit knowledge of the nurses was tested before the laboratory study. RESULTS: No systematic differences in explicit knowledge could be observed, i.e. differences in performance could not be attributed to this knowledge mode. Results from multidimensional scaling procedures illustrate differences in the tacit knowledge of nurses who successfully accomplished the critical situation and those who did not. CONCLUSIONS: The findings are in line with the assumption that experience-guided working is of the utmost importance for dealing with critical situations. Consequences of these results for nursing and person-related services in general are discussed and the aim of future research is outlined. PMID- 11380738 TI - Development of a patient-orientated instrument to measure service quality in outpatient departments. AB - AIMS OF THE STUDY: To describe three stages in the development of an instrument to measure service quality from the patients' perspective in hospital outpatient departments. RATIONALE: A reliable and valid service quality instrument is a valuable tool for collecting feedback from patients when improving outpatient services. DESIGN: A multiphase process with several versions of the questionnaire testing its validity and reliability. The first version of the questionnaire was constructed from information collected in a previous interview study of patients' service experiences. Using this version (47 questions) information was collected in 15 outpatient departments (314 patients) and the second version of the questionnaire was developed. At this stage, a survey of employees (n=111) was included in the development process. This second version of the questionnaire (43 questions) was used to collect more data in three outpatient departments of a university hospital and in two outpatient departments of two regional hospitals on three different occasions (1416, 369 and 124 patients) within a period of 2 years. The reliability of the instrument was tested in terms of stability by using three repeated measurements, and using the Cronbach's alpha coefficient as an indicator of internal consistency. Content validity was assessed by means of percentage agreement between staff and patients. Logistic regression analysis was performed to assess construct validity. RESULTS: The final version of the questionnaire contained 12 items. Agreement between patient and staff ratings was found to be acceptable for most questions (content validity). Two measurements on different occasions yielded a similar structure (construct validity). Internal consistency was acceptable (Cronbach's alpha = 0.67-0.93 in the first and 0.71 0.94 in the second survey). The variance of the alpha coefficients was small in the retest (stability). CONCLUSIONS: The instrument developed is general to the extent that it is suitable for assessing service quality improvement needs in individual units and for making cross-departmental comparisons. PMID- 11380739 TI - Swedish nurses' attitudes towards research and development within nursing. AB - AIM OF THE STUDY: The aim was to develop and test an assessment instrument in order to study attitudes towards research and development within nursing among randomly selected professional nurses, registered nurses (RNs). BACKGROUND: The investigation was initiated because of the development within nursing and nursing education in Sweden towards better knowledge about research for the last four decades. METHODS: A questionnaire was designed, consisting of three parts: (1) demographic data, (2) attitude scale and (3) research awareness. Appropriate psychometric statistics were used such as factor analysis and parametric as well as nonparametric statistics to compare groups. The form was distributed to 407 randomly selected RNs. RESULTS: The response rate was 71% (n=289). The respondents were from four different examination years, representing four different nursing education systems in Sweden. Factor analysis (Maximum Likelihood and Oblimin rotation) gave seven factors with a total variance of 58% and Cronbach's alpha between 0.60 and 0.84. The factors were labelled 'Research language', 'Need of research knowledge', 'Participation', 'The Profession', 'Meaningfulness', 'Study literature' and 'Developing - Resources'. The results indicated that the respondents in general had positive attitudes towards nursing research. There were significant differences between the examined groups. The group examined in 1966 appeared as the least positive group. Forty-six per cent never or seldom utilized nursing research findings in their daily practice. About half of the respondents never read research reports. CONCLUSION: The results indicated that the new instrument is sensitive for measuring professional nurses' attitudes towards research and development in nursing. Even if the respondents had a positive attitude towards research and development, there was a poor application in their daily work. Further, the respondent's age, the year of RN examination and acquisition of research skills seemed to be of importance for the attitudes. PMID- 11380741 TI - Visual risk factors for falls in older people. AB - OBJECTIVES: To determine the tests most predictive of falls in community-dwelling older people from a range of visual screening tests (high and low contrast visual acuity, edge contrast sensitivity, depth perception, and visual field size). To determine whether one or more of these visual measures, in association with measures of sensation, strength, reaction time, and balance, can accurately predict falls in this group. DESIGN: Prospective cohort study of 12 months duration. SETTING: Falls and Balance Laboratory, Prince of Wales Medical Research Institute. PARTICIPANTS: 156 community-dwelling men and women age 63 to 90 (mean age 76.5, standard deviation = 5.1). MEASUREMENTS: Screening tests of vision, sensation, strength, reaction time and balance, falls. RESULTS: Of the 148 subjects available at follow-up, 64 (43.2%) reported falling, with 32 (21.7%) reporting multiple falls. Multiple fallers had decreased vision, as indicated by all visual tests, with impaired depth perception, contrast sensitivity, and low contrast visual acuity being the strongest risk factors. Subjects with good vision in both eyes had the lowest rate of falls, whereas those with good vision in one eye and only moderate or poor vision in the other eye had elevated falling rates-equivalent to those with moderate or poor vision in both eyes. Discriminant analysis revealed that impaired depth perception, slow reaction time, and increased body sway on a compliant surface were significantly and independently associated with falls. These variables correctly classified 76% of the cases, with similar sensitivity and specificity. CONCLUSION: The study findings indicate that impaired vision is an important and independent risk factor for falls. Adequate depth perception and distant-edge-contrast sensitivity, in particular, appear to be important for maintaining balance and detecting and avoiding hazards in the environment. PMID- 11380742 TI - Reducing delirium after hip fracture: a randomized trial. AB - OBJECTIVES: Delirium (or acute confusional state) affects 35% to 65% of patients after hip-fracture repair, and has been independently associated with poor functional recovery. We performed a randomized trial in an orthopedic surgery service at an academic hospital to determine whether proactive geriatrics consultation can reduce delirium after hip fracture. DESIGN: Prospective, randomized, blinded. SETTING: Inpatient academic tertiary medical center. PARTICIPANTS: 126 consenting patients 65 and older (mean age 79 +/- 8 years, 79% women) admitted emergently for surgical repair of hip fracture. MEASUREMENTS: Detailed assessment through interviews with patients and designated proxies and review of medical records was performed at enrollment to ascertain prefracture status. Subjects were then randomized to proactive geriatrics consultation, which began preoperatively or within 24 hours of surgery, or "usual care." A geriatrician made daily visits for the duration of the hospitalization and made targeted recommendations based on a structured protocol. To ascertain study outcomes, all subjects underwent daily, blinded interviews for the duration of their hospitalization, including the Mini-Mental State Examination (MMSE), the Delirium Symptom Interview (DSI), and the Memorial Delirium Assessment Scale (MDAS). Delirium was diagnosed using the Confusion Assessment Method (CAM) algorithm. RESULTS: The 62 patients randomized to geriatrics consultation were not significantly different (P>.1) from the 64 usual-care patients in terms of age, gender, prefracture dementia, comorbidity, type of hip fracture, or type of surgical repair. Sixty-one percent of geriatrics consultation patients were seen preoperatively and all were seen within 24 hours postoperatively. A mean of 10 recommendations were made throughout the duration of the hospitalization, with 77% adherence by the orthopedics team. Delirium occurred in 20 /62 (32%) intervention patients, versus 32 / 64 (50%) usual-care patients (P =.04), representing a relative risk of 0.64 (95% confidence interval (CI) = 0.37-0.98) for the consultation group. One case of delirium was prevented for every 5.6 patients in the geriatrics consultation group. There was an even greater reduction in cases of severe delirium, occurring in 7/ 60 (12%) of intervention patients and 18 / 62 (29%) of usual-care patients, with a relative risk of 0.40 (95% CI = 0.18-0.89). Despite this reduction in delirium, length of stay did not significantly differ between intervention and usual-care groups (median +/- interquartile range = 5 +/- 2 days in both groups), likely because protocols and pathways predetermined length of stay. In subgroup analyses, geriatrics consultation was most effective in reducing delirium in patients without prefracture dementia or activities of daily living (ADL) functional impairment. CONCLUSIONS: Proactive geriatrics consultation was successfully implemented with good adherence after hip-fracture repair. Geriatrics consultation reduced delirium by over one-third, and reduced severe delirium by over one-half. Our trial provides strong preliminary evidence that proactive geriatrics consultation may play an important role in the acute hospital management of hip-fracture patients. PMID- 11380743 TI - A nurse-led interdisciplinary intervention program for delirium in elderly hip fracture patients. AB - OBJECTIVES: To develop and test the effect of a nurse-led interdisciplinary intervention program for delirium on the incidence and course (severity and duration) of delirium, cognitive functioning, functional rehabilitation, mortality, and length of stay in older hip-fracture patients. DESIGN: Longitudinal prospective before/after design (sequential design). SETTING: The emergency room and two traumatological units of an academic medical center located in an urban area in Belgium. PARTICIPANTS: 60 patients in an intervention cohort (81.7% females, median age = 82, interquartile range (IQR) = 13) and another 60 patients in a usual care/nonintervention cohort (80% females, median age = 80, IQR = 12). INTERVENTION: (1) Education of nursing staff, (2) systematic cognitive screening, (3) consultative services by a delirium resource nurse, a geriatric nurse specialist, or a psychogeriatrician, and (4) use of a scheduled pain protocol. MEASUREMENTS: All patients were monitored for signs of delirium, as measured by the Confusion Assessment Method (CAM). Severity of delirium was assessed using a variant of the CAM. Cognitive and functional status were measured by the Mini-Mental State Examination (MMSE) (including subscales of memory, linguistic ability, concentration, and psychomotor executive skills) and the Katz Index of activities of daily living (ADLs), respectively. RESULTS: Although there was no significant effect on the incidence of delirium (23.3% in the control vs 20.0% in the intervention cohort; P =.82), duration of delirium was shorter (P =.03) and severity of delirium was less (P =.0049) in the intervention cohort. Further, clinically higher cognitive functioning was observed for the delirious patients in the intervention cohort compared with the nonintervention cohort. Additionally, a trend toward decreased length of stay postoperatively was noted for the delirious patients in the intervention cohort. Despite these positive intervention effects, no effect on ADL rehabilitation was found. Results for risk of mortality were inconclusive. CONCLUSIONS: This study demonstrated the beneficial effects of an intervention program focusing on early recognition and treatment of delirium in older hip-fracture patients and confirms the reversibility of the syndrome in view of the delirium's duration and severity. PMID- 11380744 TI - Vitamin E and lipid peroxide plasma levels predict the risk of cardiovascular events in a group of healthy very old people. AB - OBJECTIVES: To assess whether systemic oxidative stress can predict the risk of first myocardial infarction, ischemic stroke, and congestive heart failure. DESIGN: A longitudinal study started in 1992 and completed in 1997. SETTING: Community-based, outpatient. PARTICIPANTS: 102 apparently healthy, community dwelling subjects age 80 and older from the Vibrata valley, Teramo, Italy. MEASUREMENTS: Plasma vitamin E, beta-carotene, vitamin C, fluorescent products of lipid peroxidation (FPLPs), and serum lipids were determined at enrollment. RESULTS: Thirty-two cardiovascular events were recorded in 47.4 months of follow up. The subjects with vitamin E levels in the highest quartile had a risk of cardiovascular events one-sixth those with vitamin E levels in the lowest quartile (relative risk (RR) = 0.16; 95% confidence interval (CI) = 0.04-0.55). The subjects with FPLPs in the highest quartile had a risk seven times greater than those with FPLPs in the lowest quartile (RR = 7.61; 95% CI = 2.23-25.96). No association was observed for vitamin C, beta-carotene, or total cholesterol. Multivariate adjustment for known risk factors did not significantly change the results. CONCLUSIONS: Our results suggest that in apparently healthy, community dwelling very old subjects, base-line plasma concentration of vitamin E and FPLPs predicts the risk of future cardiovascular events. We confirm previous data showing that total cholesterol is not a predictor of cardiovascular disease in people age 80 and older. PMID- 11380745 TI - Ethnic differences in mini-mental state examination (MMSE) scores: where you live makes a difference. AB - OBJECTIVES: To examine differences in correlates of the Mini-Mental State Examination (MMSE) in a population-based sample of older Mexican Americans and European Americans and to provide empirical validation of the MMSE as an indicator of cognitive impairment in survey research in older Mexican Americans by comparing MMSE classification against performance on timed tasks with varying levels of cognitive demand. DESIGN: A population-based cross-sectional study. SETTING: Trained bilingual staff administered the MMSE as part of the San Antonio Longitudinal Study of Aging (SALSA) home-based assessment battery. PARTICIPANTS: 827 community-dwelling Mexican Americans and European Americans, 65 and older, residing in three socioeconomically and culturally distinct neighborhoods (barrio, transitional, suburban). MEASUREMENTS: The MMSE was compared against a variety of demographic, biomedical, and sociocultural variables ascertained by self-report and against performance-based measures of functional tasks representing varying levels of cognitive demand (Structured Assessment of Independent Living Skills (SAILS) subscales for food manipulation and money management). RESULTS: Mexican Americans were 2.2 times more likely than European Americans to have MMSE scores <24. Multiple logistic regression analysis revealed that neighborhood was an independent predictor of low MMSE scores in Mexican Americans, with the relationship between ethnic group and MMSE explained by neighborhood. After adjusting for neighborhood type, no differences were noted between Mexican Americans and European Americans. Independent of other factors examined, low education was associated with low MMSE scores in both Mexican Americans and European Americans. Mexican Americans with MMSE scores <24 took significantly longer to complete four out of five performance-based food manipulation tasks and all three money management tasks. CONCLUSIONS: Neighborhood type was a predictor of cognitive impairment. Education affected MMSE scores similarly in both ethnic groups. MMSE scores <24, indicative of cognitive impairment, were uniformly associated with functional impairment in both the Mexican Americans and European Americans. Among older Mexican Americans, MMSE-classified cognitive impairment was significantly associated with poorer performance on timed tasks with varying levels of cognitive demand independent of other correlates. A similar pattern of association was observed in European Americans. Thus, the MMSE appears to be a valid indicator of cognitive impairment in survey research in both older Mexican Americans and European Americans. PMID- 11380746 TI - Physician visits, emergency room utilization, and overnight hospitalization in the old-old in Israel: the cross-sectional and longitudinal aging study (CALAS). AB - OBJECTIVES: The objective of this paper is to assess the risk factors for physician contact in the month before the interview (PM) and emergency room utilization (ERU) and overnight hospitalization (OH) in the year before the interview, through the use of the behavioral model as a conceptual framework. DESIGN: A random stratified sample of subjects age 75 to 94 was selected from the National Population Register (a complete listing of the Israeli population maintained by the Ministry of the Interior). The study sample consisted of Jews living in Israel on January 1, 1989, stratified by age (four 5-year age groups: 75-79, 80-84, 85-89, and 90-94), sex, and place of birth (Europe/America, Asia/Africa, and Israel). SETTING: Community-dwelling old-old Jewish Israelis. PARTICIPANTS: 1,487 people living in the community at the time of the baseline interview. MEASUREMENTS: The dependent variables were PM in the month before the interview and ERU and OH in the previous year. The independent variables were: predisposing variables (age, sex, place of birth, and education); enabling variables (income and the social network variables of marital status, living arrangements, and number of in-person contacts per week with any child); and need variables (number of self-reported chronic medical conditions, subjective health, depressive symptoms, number of difficulties with activities of daily living and instrumental activities of daily living, measures of physical robustness, and engaging in regular physical sportive activities). RESULTS: The predisposing and enabling factors were only minimally associated with utilization rates in the old old in Israel, with the exception of lower rates of ERU by those who were living alone. Age was not significantly associated with healthcare utilization in the old-old population studied. Healthcare utilization was found to be associated primarily with health and functional status. CONCLUSION: In a system of free and equal access to healthcare services, the demand for health services by a population with high levels of chronic disease and disability is driven primarily by health needs, rather than by extraneous factors such as income and education. The study indicates that equity in the provision of health services is attainable. Policy makers should provide for actual need, remove artificial barriers, and prepare accurate estimates of future needs. PMID- 11380747 TI - Aspiration pneumonia: dental and oral risk factors in an older veteran population. AB - OBJECTIVES: To investigate the importance of medical and dental factors in aspiration pneumonia in an older veteran population. DESIGN: Prospective enrollment of subjects with retrospective analysis of data. SETTING: Department of Veterans Affairs outpatient clinic, inpatient ward, and nursing home. PARTICIPANTS: 358 veterans age 55 and older; 50 subjects with aspiration pneumonia. MEASUREMENTS: Demographic and medical data; functional status; health related behaviors; dental care utilization; personal oral hygiene; comprehensive dental examination; salivary assays including IgA antibodies; and cultures of saliva, throat, and dental plaques. RESULTS: Two logistic regression models produced estimates of significant risk factors. One model using dentate patients included: requiring help with feeding (odds ratio (OR) = 13.9), chronic obstructive pulmonary disease (COPD) (OR = 4.7), diabetes mellitus (OR = 3.5), number of decayed teeth (OR = 1.2), number of functional dental units (OR = 1.2), presence of important organisms for decay, Streptococcus sobrinus in saliva (OR = 6.2), and periodontal disease, Porphyromonous gingivalis in dental plaque (OR = 4.2), and Staphylococcus aureus presence in saliva (OR = 7.4). The second model, containing both dentate and edentulous patients included: requiring help with feeding (OR = 4.7), COPD (OR = 2.5), diabetes mellitus (OR = 1.7), and presence of S. aureus in saliva (OR = 8.3). CONCLUSION: This study supports the significance of oral and dental factors while controlling for established medical risk factors in aspiration pneumonia incidence. PMID- 11380748 TI - Outcome of older patients requiring ventilatory support in intensive care: impact of nutritional status. AB - OBJECTIVES: To determine predictors of mortality in the intensive care unit (ICU) and at 6 months after discharge; to assess the lifestyles of survivors 6 months after discharge. DESIGN: Prospective cohort study of patients screened upon admission and 6 months after discharge from the ICU. SETTING: The ICU of a university hospital. PARTICIPANTS: One hundred sixteen consecutive patients age 70 and older admitted to the ICU and treated by mechanical ventilation for at least 24 hours. MEASUREMENTS: A comprehensive medical, functional, nutritional, and social assessment was undertaken for each patient upon admission to the ICU. Functional status and residence were recorded for patients still living 6 months after discharge from the ICU. RESULTS: Mortality in the ICU and 6 months after discharge was 31% and 52%, respectively. The predictors of in-ICU mortality on multivariate analysis were a high omega score per day in the ICU and a high simplified acute physiologic score corrected for points related to age (SAPS IIc). The predictors of mortality at 6 months were a high omega score per day in the ICU, a high SAPS IIc, and a mid-arm circumference (MAC) under the 10th percentile for the older French population in good health. Six months after discharge from the ICU, 91% of the surviving patients had the same residential status and 89% had a similar or improved functional status compared with pre admission status. CONCLUSIONS: Although severity of illness remains an important predictor of in-ICU mortality and mortality at 6 months after release from ICU, we found that impaired nutritional status upon admission was related to 6-month mortality. These results emphasize the need for a systematic nutritional assessment in older patients admitted to the ICU and treated by mechanical ventilation. PMID- 11380749 TI - Cause of death in older patients with anatomo-pathological evidence of chronic bronchitis or emphysema: a case-control study based on autopsy findings. AB - OBJECTIVES: To determine the most frequent causes of death of hospitalized older patients based on anatomo-pathological evidence and to compare the relative frequency of fatal events between patients with and without evidence of either chronic bronchitis (CB) or emphysema (E). DESIGN: Retrospective, case-control study based on a computerized database including anatomo-pathological data of patients deceased and autopsied over a 25-year period. SETTING: Two geriatric hospitals in Geneva. PARTICIPANTS: Not applicable. MEASUREMENTS: Autopsy records for cause(s) of death in patients with CB or E. RESULTS: 3,685 patients deceased in our institution (1,540 men; 2,145 women) were autopsied between 1972 and 1996; mean age at death was 81.5 +/- 8.0 years. Anatomo-pathological evidence of CB or E was found in 983 patients (26.6% of total); 262 (7.2%) had predominantly CB, and 456 (12.3%) predominantly E. Pneumonia was the most frequent cause of death in all patients (21.8%). Myocardial infarction (MI) (17.6% vs 14%), and respiratory failure (5.1% vs 1.5%) occurred more frequently in subjects with CB and/or E than in controls. Fatal pulmonary embolism (PE) was more frequent in patients with E (18.4%) than in patients with CB (10.7%; odds ratio ( OR) = 1.89, P =.008), or in controls (12.7%; OR = 1.56, P =.0008). CONCLUSION: Anatomo pathological evidence of CB or E is highly prevalent in older patients, suggesting that CB and E are clinically underdiagnosed in this age group. Fatal MI occurred significantly more frequently in older patients with E or CB than in controls. Furthermore, patients with E were at significantly higher risk of fatal PE than patients with CB or controls. PMID- 11380750 TI - Effect of nonsteroidal anti-inflammatory drug use on the rate of gastrointestinal hospitalizations among people living in long-term care. AB - OBJECTIVES: Gastrointestinal (GI) complications are the most-common serious adverse reactions associated with nonsteroidal anti-inflammatory drugs (NSAIDs). We quantified the effect of specific NSAIDs on the rate of GI hospitalizations among older people living in long-term care. DESIGN: Retrospective cohort study. SETTING: All Medicare/Medicaid certified nursing homes in four states (Maine, Minnesota, New York, and South Dakota). PARTICIPANTS: We identified 125,516 newly admitted residents from a database of all residents (1992-1996) of all Medicare/Medicaid certified nursing homes in four states. Using the federally mandated Minimum Data Set, which includes information on all drugs received (prescription and over-the-counter), we identified patients who received at least one prescription for aspirin (n = 19,101) or NSAIDs (n = 9,777). The control population consisted of all institutionalized persons who did not receive these drugs. MEASUREMENTS: From Health Care Financing Administration inpatient claims, we identified the first hospitalization for GI perforation, ulcer, or hemorrhage that occurred during the year of follow up (ICD9-CM discharge codes: 531-534, 578). Cox proportional hazards models provided adjusted estimates of rate ratios. RESULTS: NSAID exposure increased the GI-event-related hospitalization rate in both men (rate ratios (RR) = 2.64; 95% confidence interval (CI) = 1.17-5.99) and women (RR = 3.23; 95% CI = 1.85-5.65). The rate of GI hospitalizations for both men and women taking sulindac, naproxen, or indomethacin was higher than for nonusers. The risk of GI-event-related hospitalizations was greatest among women exposed to diflunisal (RR = 6.08; 95% CI = 2.27-16.26) or oxaprozin (RR = 6.03; 95% CI = 2.49-14.58). CONCLUSIONS: Despite the high background rate of GI events, most NSAIDs increased the risk of GI hospitalization. Careful attention to choice of agent and dosing is needed in prescribing NSAIDs in this frail, older population. PMID- 11380751 TI - Prevalence of tooth loss and dental service use in older Mexican Americans. AB - OBJECTIVES: To describe the prevalence of tooth loss, to examine risk factors for having fewer teeth or no teeth, and to describe the use of dental services in an older Mexican-American population. DESIGN: Data from the baseline phase of the Hispanic Established Population for the Epidemiological Study of the Elderly survey conducted from 1993 to 1994, a cross-sectional survey of older Mexican Americans. SETTING: Five southwestern states: Texas, California, Arizona, New Mexico, and Colorado. PARTICIPANTS: 3,050 noninstitutionalized Mexican Americans age 65 to 99. RESULTS: Twenty-seven percent of the sample was completely edentulous and 22% reported visiting or speaking with a dental care professional in the preceding year. Logistic regression analyses showed that being older or being female was significantly associated with tooth loss, adjusting for education, income, smoking status, and diabetes mellitus. Current smokers (odds ratio (OR) = 1.69; 95% CI = 1.31-2.20) and diabetics (OR = 1.53; 95% CI = 1.27 1.84) were more at risk for tooth loss, as were persons of lower socioeconomic status. CONCLUSIONS: The prevalence of tooth loss and use of dental services in this population of older Mexican Americans is lower than what has been previously found among older people in the general population. PMID- 11380752 TI - Problem behavior in the last year of life: prevalence, risks, and care receipt in older Americans. AB - OBJECTIVES: To estimate the prevalence of problem behavior in the last year of life in older people and to explore risk factors and assess the effect of behaviors on access to care. DESIGN: Retrospective analysis of data from the 1993 National Mortality Followback Survey, conducted by the National Center for Health Statistics (NCHS). SETTING: Persons who resided and died in the United States (except South Dakota) in 1993. PARTICIPANTS: Seven thousand six hundred and eighty-four deaths in people age 65 and older were included, from which full informant interview data were available for 6,748 decedents (88%). MEASUREMENTS: Informant data were collected on frequency of complaints about behavior from family members, complaints from others in the community, bizarre behavior, destroying property, violent threats or attempts, and temper tantrums. RESULTS: Overall, 20% of decedents were reported as having any of the problem behaviors sometimes or often in the last year of life. Rates differed little by age at death or gender. Risks of having problem behaviors were higher for those with clinically diagnosed dementia, mental illness, alcohol abuse, and bronchitis or emphysema. A diagnosis of dementia had been made in 27% of those with behavior problems. Nursing homes or healthcare facilities were the usual residence of 32% of people with any behavior problems sometimes or often during their last year of life. Informants for decedents who had destroyed property or made violent threats were 2.3 times (95% confidence interval (CI) = 1.2-4.4) more likely to report that the subject had not received the care they had needed during the last year of life. CONCLUSION: Problem behavior is relatively common in older people in the last year of life and is not confined to nursing home residents or people suffering from dementia. PMID- 11380753 TI - Secular trends in self-reported functioning, need for assistance and attitudes towards life: 10-year differences of three older cohorts. AB - OBJECTIVES: To compare the self-reported functional status of cohorts, born 10 years apart, when they were at equivalent ages: 75, 80, or 85. DESIGN: Cross sectional mailed survey of three birth cohorts in 1989 and 1999. PARTICIPANTS: Random samples of older home-dwelling residents from birth cohorts 1904, 1909, and 1914 in 1989 ( N = 685) and the birth cohorts 1914, 1919, and 1924 in 1999 ( N = 2,047) were asked the same questions. MEASUREMENTS: Self-reported physical functioning, need for assistance in daily living, and attitudes toward life. RESULTS: Among 85-year-olds born in 1914 there was a significantly larger proportion able to go outdoors compared with 85-year-olds born in 1904 (72.9% vs 60.6% in women (P <.01) and 84.6% vs 63.6 % in men (P <.01), respectively). Similar trends were observed between the oldest cohorts concerning the need for assistance. The amount of publicly funded domestic help had reduced in the two oldest cohorts in 1999 compared with 1989 (20.3% vs 29.8% in 85-year-old women born in 1914 vs 1904 (P <.05); and 10.2 % vs 25.0% in 85-year-old men born 1914 vs 1904 (P <.05), but at the same time 75-year-old women born in 1924 had increased the use of private domestic help compared with 75-year-old women born in 1914. Significantly larger proportions of both men and women had plans for the future in all the cohorts in 1999 than in 1989. CONCLUSIONS: Consistent yet small differences between the corresponding cohorts suggest that physical functioning and more-optimistic attitude toward life have increased and need for assistance has decreased over 10 years in the older population up to the age of 85. PMID- 11380754 TI - Functional status and self-rated health in 2,262 nonagenarians: the Danish 1905 Cohort Survey. AB - OBJECTIVES: To describe the functional capacity and self-rated health of a large cohort of nonagenarians. DESIGN: A cross-sectional survey of all Danes born in 1905 (92-93 years of age), carried out August to October 1998. SETTING: Participants' homes. PARTICIPANTS: Two thousand two hundred and sixty-two nonagenarians, corresponding to a participation rate of 63% (of these, 20% participated by proxy). MEASUREMENTS: Activities of daily living (ADLs) and self rated health were assessed by interview. Five items from Katz's ADLs (bathing, dressing, transfer, toileting, and eating) were used to construct a three-level five-item ADL scale (not disabled (no disabilities), moderately disabled (1-2 disabilities), severely disabled (3-5 disabilities)). From responses to a more extensive list of questions on ADLs (26 items), we identified scales of strength and agility by means of factor analysis. Furthermore, a 26-item ADL scale was made. Physical performance tests (chair stand, timed walk, lifting a 2.7 kg box, maximum grip-strength, and flexibility tests) were performed among nonproxy responders. RESULTS: According to the five-item ADL scale, 50% of the men and 41% of the women were categorized as not disabled, while 19% and 22%, respectively, were categorized as severely disabled. The five-item ADL scale correlated highly with the 26-item ADL scale (r = 0.83). The ADL scales showed moderate-to-good correlation with each other (r = 0.74-0.83), and with the physical performance tests (r = 0.31-0.58). Only 3.7% of the women and 6.3% of the men walked (normal pace) with a speed of at least 1 meter per second, which is the minimum walking speed required to cross signaled intersections in Denmark. A total of 56% considered their health to be excellent or good. Of the participants, 74% were always or almost always satisfied with their lives, even though only 45% reported that they "felt well enough to do what they wanted." The analyses showed that no single ADL item seemed to be of particular importance for how the participants rated their health. CONCLUSION: The Danish 1905 cohort survey is the largest and the only nationwide survey of a whole birth-cohort of nonagenarians. A total of 2,262 fairly nonselected nonagenarians participated. The level of both self reported disability and functional limitations measured by physical performance tests among nonagenarians was high. Despite their lower mortality, women were more disabled than men and did not perform as well as men in the physical performance tests. Nevertheless, the majority of the participants considered their health to be good and were satisfied with their lives. PMID- 11380755 TI - ACE gene polymorphism and insulin action in older subjects and healthy centenarians. AB - OBJECTIVES: To evaluate the possible relationship between angiotensin-converting enzyme (ACE) insertion-deletion (ID) genotype and insulin resistance in a population of healthy older Italian subjects. DESIGN: Prospective recruitment of a convenience sample. PARTICIPANTS: One hundred twenty-five subjects age 62 to 105 in good health and not taking any drug known to interfere with glucose metabolism. RESULTS: In the sample population, the relative frequencies of the ACE genotypes deletion-deletion (DD) (0.424), ID (0.400), and insertion-insertion (II) (0.176) were not significantly different from values predicted by Hardy Weinberg equilibrium. The genotype distribution was similar in men and women. Subjects carrying the II genotype had a higher FPG (P <.001) and FPI (P <.001) than did subjects with DD or ID genotype. Subjects with II genotype also had a significantly higher HOMA index than did subjects with DD or ID genotype (P for trend <.002). In a multivariate stepwise regression analysis, the ACE ID polymorphism was significantly and independently associated with the HOMA index (P <.001). The same result was confirmed performing multivariate analysis in the younger group and centenarians separately. CONCLUSIONS: In an older population, the presence of II ACE genotype is associated with a high degree of insulin resistance independent of other anthropometric variables known to interfere with insulin action; this association is significant in both the younger subjects and the centenarians. PMID- 11380756 TI - Disability outcomes of older Medicare HMO enrollees and fee-for-service Medicare beneficiaries. AB - OBJECTIVES: To investigate whether older Medicare beneficiaries enrolled in Medicare risk health maintenance organizations (HMOs) have different rates of disablement than fee-for-service (FFS) beneficiaries. DESIGN: Secondary analysis of annual functional status transitions using the Medicare Current Beneficiary Survey, 1991 to 1996. SETTING: Telephone interviews. PARTICIPANTS: Forty-four thousand seven hundred and sixty-five person-years of annual functional status transitions for noninstitutionalized older Medicare beneficiaries who were either risk HMO enrollees or FFS beneficiaries with or without private supplementary insurance. MEASUREMENTS: Five multinomial logit models were estimated as single state transition models, with five functional states, death, and censored as outcomes. The probability of being in a certain functional state the following year was specified as a function of individual risk factors and HMO versus FFS supplementary insurance status. RESULTS: Among functionally independent beneficiaries, the odds of becoming disabled in activities of daily living (ADLs) within a year were lower among FFS individuals with supplementary insurance (odds ratios (OR) = 0.67, P <.01) and HMO enrollees (OR = 0.58, P <.01). Among older people who were functionally impaired, neither HMO enrollment nor private supplementary insurance affected the risk of further functional decline or functional improvement. Supplementary insurance, but not HMO enrollment, was associated with lower mortality risk among beneficiaries with functional limitations (OR = 0.65, P <.05) or moderate ADL disability (OR = 0.72, P <.05). CONCLUSION: Medicare risk HMO enrollment and FFS private supplementary insurance convey similar benefits of slowing functional decline and extending life span for nonseverely disabled older people. That no association was found between adverse functional status outcomes and risk HMO enrollment has favorable implications regarding the quality of care of managed care plans. PMID- 11380757 TI - A self-paced step test to predict aerobic fitness in older adults in the primary care clinic. AB - OBJECTIVES: To study the potential usefulness of a submaximal self-paced step test as a prediction of maximal aerobic capacity (VO2max) in older adults in the primary care setting. DESIGN: Data were collected during a prospective randomized study of an exercise program. SETTING: Four university family medical clinics in London, Ontario, Canada. PARTICIPANTS: A random sample of 240 healthy older (> or =65) men (n = 118) and women (n = 122) from four family medical clinics underwent self-paced step testing in the clinic with a family physician (n = 16), and step testing and a maximal exercise treadmill test with measurement of respired gases in an exercise laboratory. Testing was done in random order (clinic/laboratory) separated by 2 weeks and then repeated at 52 weeks, following introduction of an exercise program. Relationships between outcome variables were examined by Pearson correlation coefficients while prediction of VO2max was examined using multivariate regression analysis. Cross-validation with 30 age-matched hypertensive and 40 age-matched post-hip arthroplasty patients was used to test the accuracy of the predictive models. MEASUREMENTS: Measured VO2max, predicted VO2max, step test time, step test heart rate, body mass index (BMI), and O2 pulse. RESULTS: Two hundred women (n = 108) and men (n = 92) completed both the initial and 52-week assessments. Stepping time, heart rate, age, BMI, and O2 pulse were strongly associated with VO2max for both a normal and a fast step pace and were chosen to develop the predictive model. Normal step-pace correlation with VO2max (ml/kg/min) was no different (female 0.93: male 0.91) from fast pace (0.95:0.90) with no difference between clinic and laboratory measurement at baseline or 52 weeks. Cross-validation showed no significant difference from the main group using the predictive model. CONCLUSIONS: The self-paced step test is a safe and simple clinical instrument that strongly and reliably predicts VO2max, is sensitive to change, and is generalizable in the family practice setting among community-dwelling older adults differing in fitness and health status. PMID- 11380758 TI - Sleep disturbances and mortality: results from the Canadian Study of Health and Aging. AB - OBJECTIVES: To cross-validate, in a secondary analysis, the observation that daytime sleepiness is associated with an increased risk of death. DESIGN: Prospective cohort study. SETTING: Canada, a nationally representative sample of people age 65 and older. PARTICIPANTS: Nine thousand and eight community-dwelling participants in the Canadian Study of Health and Aging. MEASUREMENTS: EXPOSURES: self-reported sleep disturbances. OUTCOMES: Cox hazard ratios (HRs) for death. RESULTS: The unadjusted analysis showed a small increased risk of death from daytime sleepiness (HR = 1.89; 95% confidence interval (CI) = 1.44-2.46), but this finding did not persist in a multivariate model adjusted for age, depression, cognition, comorbid illness, and function. CONCLUSION: Daytime sleepiness itself is not associated with an increased risk of death when other factors are taken into account. Daytime sleepiness may be a proxy for other morbid conditions and therefore for overall tiredness. PMID- 11380759 TI - Serum carotenoids and cerebral white matter lesions: the Rotterdam scan study. AB - OBJECTIVES: To study the relation between serum levels of carotenoids and white matter lesions (WMLs) on magnetic resonance imaging (MRI). DESIGN: Evaluation of cross-sectional data from a cohort study. SETTING: The Rotterdam Scan Study. PARTICIPANTS: Two hundred and three nondemented older persons, age 60 to 90, from the Rotterdam Scan Study. MEASUREMENTS: Serum levels of carotenoids were determined. WMLs on MRIs were rated separately into periventricular and subcortical WMLs. Odds ratios (ORs) for the presence of severe WMLs (upper decile) were calculated per standard deviation (SD) increase in serum carotenoid level and per SD increase in overall carotenoid serum level. Effect modification by smoking status was studied through stratified analyses. RESULTS: Increasing levels of all the separate carotenoids were associated with less severe periventricular WMLs, which reached statistical significance for the overall carotenoid serum level (OR 0.4 per SD; 95% confidence interval (CI) = 0.2-0.9). We found no association between carotenoid levels and the presence of severe subcortical WMLs (OR 1.2 per SD; 95% CI = 0.7-2.0). The association of carotenoid levels with severe periventricular WMLs was more marked in those who ever smoked (OR 0.1 per SD; 95% CI = 0.0-0.9) than in those who had never smoked (OR 0.9 per SD; 95% CI = 0.4-2.1). CONCLUSIONS: These findings are compatible with the view that high levels of carotenoids may protect against WMLs in the periventricular region, in particular in smokers. Longitudinal studies with repeated measurements of both carotenoids and WMLs are necessary to explore this hypothesis further. PMID- 11380760 TI - Urban-rural differences in a memory disorders clinical population. AB - OBJECTIVES: To compare patient characteristics and family perceptions of patient function at one urban and one rural memory disorders clinic. DESIGN: Secondary, cross-sectional data analyses of an extant clinical database. SETTING/PARTICIPANTS: First time visits (n = 956) at two memory disorders clinics. MEASUREMENTS: Patient and family-member demographics and assessment results for the Mini-Mental State Examination (MMSE), instrumental activities of daily living (IADLs), activities of daily living (ADLs), the Memory Change and Personality Change components of the Blessed Dementia Rating Scale, and the Revised Memory and Behavior Problems Checklist. RESULTS: In both clinics, patients and family members were more likely female. The typical urban clinic patient was significantly more likely to be living in a facility and more educated than the typical rural patient. Urban and rural patients did not show significant differences in age- and education-adjusted MMSE scores or raw ADL/IADL ratings, but the urban family members reported more memory problems, twice as many personality changes, more-frequent behavior problems, and more adverse reactions to problems. CONCLUSION: Physicians who practice in both urban and rural areas can anticipate differences between patients, and their families, who seek a diagnosis of memory disorders. Our most important finding is that despite similarities in reported functional abilities, urban families appear to be more sensitive to and more distressed by patients' cognitive and behavioral symptoms than rural families. These differences may reflect different underlying needs, and should be explored in further research. PMID- 11380761 TI - High vitamin E plasma levels and low low-density lipoprotein oxidation are associated with the absence of atherosclerosis in octogenarians. AB - OBJECTIVES: To identify the biological characteristics of older subjects with vascular successful aging (VASA), defined as the absence of instrumental signs and clinical symptoms of atherosclerosis in the extracoronary and coronary vessels. DESIGN: A cross-sectional study. SETTING: A university-affiliated outpatient clinic. PARTICIPANTS: Sixty older subjects (30 with VASA and 30 controls with moderate carotid atherosclerosis (AG group)) from a sample of 705 subjects age 75 and older consecutively screened. MEASUREMENTS: Clinical examination; ultrasonographic examination of carotid, vertebral, abdominal aortic, iliac, and femoral arteries; electrocardiogram; and laboratory evaluation (lipid profile, lipophilic antioxidants, and markers of low-density lipoprotein (LDL) oxidation). RESULTS: Compared with controls, there were more females in the VASA group (82% vs 50%, P <.01), and fewer previous smokers (20.5% vs 52.5%, P <.01). Vitamin E/total cholesterol levels both in plasma (4.81 vs 3.51 micromol/mmol, P <.001) and in isolated LDLs (2.71 vs 1.86 microg/mg LDL cholesterol (LDL-C), P <.01), were higher in the VASA group, as was the resistance of LDLs to in vitro oxidation (as indicated by a longer duration of the lag phase: 80.2 vs 65.6 minutes, P <.001). The level of fluorescent products of lipid peroxidation (FPLPs) in native LDLs was lower in the VASA group (13.5 vs 18.8 URF/mg LDL-C, P <.001). Multivariate logistic regression analysis showed that only plasma vitamin E level (odds ratio (OR) = 6.04, 95% confidence interval (CI) = 1.48-24.63) and FPLPs in LDLs (OR = 0.53, 95% CI = 0.31-0.91) were independently associated with VASA. CONCLUSIONS: These results suggest that an appropriate level of vitamin E and a low level of LDL oxidation might be important for reaching advanced age without developing atherosclerosis. PMID- 11380762 TI - Screening for cervical carcinoma in older women. American Geriatrics Society. PMID- 11380763 TI - Laparoscopic surgery in older adults. AB - This article provides an overview of the current role of laparoscopic surgery in older patients. A retrospective review and analysis of the recent English language literature on laparoscopic procedures with special attention devoted to those articles focused on geriatric patients was performed. Laparoscopic surgery has rapidly become the fastest-growing discipline within the surgical arena and new applications for laparoscopy continue to be reported. The primary benefits to patients of these developments are smaller scars, decreased postoperative pain, and more-rapid return to normal activity. As society ages, more older patients will present with pathology amenable to laparoscopic intervention. Several aspects of laparoscopy impose unique physiologic stresses and, as such, may alter surgical risk to the geriatric patient. In addition, older patients often have delayed surgical interventions because of more-conservative medical management or unusual symptomatology, which may further complicate the laparoscopic approach. These limitations may alter the risk-to-benefit ratio of laparoscopic versus open procedures. Despite this lack of elucidation of full-risk profiles, laparoscopic approaches should be considered regardless of a patient's age. PMID- 11380764 TI - Guideline for the prevention of falls in older persons. American Geriatrics Society, British Geriatrics Society, and American Academy of Orthopaedic Surgeons Panel on Falls Prevention. PMID- 11380765 TI - Community screening for visual impairment in older people. Community screening for visual impairment in the elderly. PMID- 11380766 TI - Where is the vision for fall prevention? PMID- 11380767 TI - Delirium after hip fracture: to be or not to be? PMID- 11380768 TI - Designing a study for success. PMID- 11380769 TI - Late presentation of tethered spinal cord in a 73-year-old patient. PMID- 11380770 TI - Myasthenia gravis: an elusive diagnosis in older people. PMID- 11380771 TI - Estimation of height in older Chinese adults by measuring limb length. PMID- 11380772 TI - Dementia reversal in post-shunt normal pressure hydrocephalus predicted by neuropsychological assessment. PMID- 11380773 TI - In re "indiscriminate" dressings for pressure ulcers. PMID- 11380774 TI - Antithrombotic therapy for prevention of pneumonia. PMID- 11380776 TI - Hypocortisolemia and delirium in an older patient. PMID- 11380777 TI - Impact of an influenza pandemic on the mortality of congestive heart failure in older Japanese: the 1998 Japanese influenza pandemic. PMID- 11380778 TI - Marginal adaptation of cast partial crowns made of pure titanium and a gold alloy under influence of a manual burnishing technique. AB - The aim of this study was to determine the marginal adaptation of partial crowns from pure titanium and a gold alloy after two different cementation techniques. Forty freshly extracted human molars were prepared and randomly divided in four groups. Two groups were restored with partial crowns using the gold alloy Degulor M*. In one group, the crowns were fixed on the tooth by using a zinc phosphate cement. In the other group the margins were additionally burnished by using a hand burnisher No. 660. In the other two groups, partial crowns from pure titanium were cemented in the same way. The marginal quality was determined by quantitative margin analysis in the SEM using a replica technique. Partial crowns from a gold alloy showed significantly (P < 0.05) more margin quality A (vertical marginal discrepancy < 50 microm) while partial crowns from pure titanium had significantly (P < 0.05) more margin quality B (vertical marginal discrepancy 50 100 microm) and over-extended margins (quality D). No significant (P < 0.05) difference was found between the conventional cementation technique and the technique with manual burnishing in both material groups. PMID- 11380779 TI - An in vitro wear study of posterior denture tooth materials on human enamel. AB - This in vitro study evaluated the wear effects of five posterior denture tooth materials on human enamel. The tooth specimen was cusp shaped and enamel specimen was formed as a 10 C 10 C 5 mm plate. All material-enamel combinations were tested using a machine designed to produce sliding contact 20 C 104 times at 60 cycles min-1 and a 4-mm sliding distance per stroke in the bucco-lingual direction under a load of 1 kg. Wear analysis was measured as the total height loss of each combination. In addition, the surface roughness (Ra) of each worn specimen was also evaluated. The least total height loss occurred with poly (methyl methacrylate) (PMMA) enamel pair, and the greatest did with porcelain (Po)-enamel pair. The lowest compound Ra value was measured in high-strength resin (HR)-enamel pair, and the highest in Po-enamel pair. These findings suggest that the best combination is PMMA-enamel, and the poorest combination is Po enamel. PMID- 11380780 TI - Finite element analysis on preferable I-bar clasp shape. AB - An I-bar clasp is one of the most popular direct retainers for distal-extension removable partial dentures. However, no adequate information is available on preferable shape as determined by biomechanics. This study aimed (1) to investigate, by finite element analysis (FEA), the dimensions and stress of I-bar clasps having the same stiffness, and (2) to estimate a mechanically preferable clasp design. Three-dimensional FEA models of I-bar clasps were created with vertical and horizontal straight sections connected by a curved section characterized by six parameters: thickness of the clasp tip, width of the clasp tip, radius of the curvature, horizontal distance between the base and the vertical axis, vertical dimension between the tip and the horizontal axis, and taper (change of width per unit length along the axis). Stress was calculated with a concentrated load of 5 N applied 2 mm from the tip of the clasp in the buccal direction. A thinner and wider clasp having an taper of 0.020-0.023 and radius of curvature of 2.75-3.00 showed less stress. The results suggest that such a shape might be the preferable I-bar clasp shape as biomechanical viewpoint. PMID- 11380781 TI - Effect of sandblasting on the bond strength of the bondable molar tube bracket. AB - This study examined the effect of surface treatment with sandblasting on bracket bonding strength. Extracted human tooth, base metal alloy and porcelain surfaces were treated with sandblasting. The bracket bonding strengths of sandblasted surfaces were evaluated and compared with the controls and etched enamel surfaces. Morphological observation of the treatment surfaces and the failure sites was conducted. Statistical analysis included one-way analysis of variance (ANOVA) and a Scheffe's F-test at the 95% confidence level was performed. The results indicated that mean bond strength values ranged from 3.6 MPa (the untreated control) to 20.4 MPa for the etched enamel surface. No statistically significant differences were determined among the etched enamel, sandblasted metal and sandblasted porcelain surfaces (P > 0.05). Most debonding specimens failed at either the resin-tooth interface or within the adhesive. In conclusion, sandblasting the metal and porcelain surfaces obtained a bracket bond strength comparable with that with the etched enamel surface. PMID- 11380782 TI - An investigation into the importance of the periodontal ligament and alveolar bone as supporting structures in finite element studies. AB - Many finite element analyses studies have been published in the dental literature. Some of these have analysed just the crown of the tooth, others have included part or all of the root, while others have included the supporting periodontal ligament and alveolar bone. The aim of this study was to examine which of the supporting structures was important to the model when analysing the stress distribution within a tooth. A two-dimensional plane strain finite element model of a lower second pre-molar was developed, which included the supporting periodontal ligament and alveolar bone. Two 50 N loads were applied to simulate the effect of a load in centric occlusion. The nodal x, y and shear stresses were recorded along two horizontal planes, one in the crown and one in the cervical region. Each of the supporting structures was systematically removed and the remaining structures reanalysed. It was found that it was particularly important to include both the periodontal ligament and alveolar bone when undertaking the finite element analyses of teeth. PMID- 11380783 TI - Microwave-cured acrylic resins and silicone-gypsum moulding technique. AB - This study evaluated the residual monomer (RM), Knoop hardness (KHN) and transverse strength (TS) of two microwave-cured acrylic resins (Acron MC(R) (A), GC Dent. Ind. Corp., Tokyo, Japan and Onda Cryl (O), AO Classico Ltda, Sao Paulo, Brazil) when processed with an all-type III gypsum moulding technique (G) or a silicone-gypsum moulding technique (S). One hundred and forty four specimens were fabricated and equally distributed into four groups (AG, AS, OG and OS). The TS tests were conducted after 48 h of water storage, and KHN values were obtained after 24, 48, 72 h and 30 days. The RM was determined every 24 or 48 h over a period of 288 h. The acrylic resins were prepared and processed according to the manufacturer's directions. Both AS and OS showed the highest means (P < 0.05) for RM (microg cm-2) only after 24 h (AS = 56.84 +/- 27.39/AG = 7.51 +/- 5.75/OS = 3.59 +/- 1.60/OG = 1.02 +/- 0.3), 48 h (AS = 28.99 +/- 9.35/AG = 2.65 +/- 2.17/OS = 2.37 +/- 0.84/OG = 0.68 +/- 0.49) and 72 h (AS = 15.98 +/- 9.01/AG = 1.40 +/- 0.57/OS = 1.87 +/- 0.52/OG = 0.75 +/- 0.44). Both AS and OS showed the highest means (P < 0.02) for KH after 24 h (AS = 18.69 +/- 2.3/AG = 17.79 +/- 0.7/OS = 18.41 +/- 1.0/OG = 16.04 +/- 0.6). After 48 h the mean values for OS and OG differed significantly (P < 0.03) (OS = 18.67 +/- 0.8/OG = 16.75 +/- 0.8). No differences of KHN were found among the groups during the storage periods. The TS values for A and O were not affected by either G or S (P > 0.05). Silicone-gypsum mould technique affected the RM and KHN of the resins in the first 2 and 5 days of analysis, respectively. The type of mould did not affect TS, and the acrylic resins differed from each other for all properties regardless of the type of mould. PMID- 11380784 TI - Effects of plaque control on the patency and occlusion of dentine tubules in situ. AB - To elucidate the nature of dentine hypersensitivity, the effects of plaque control on the patency and occlusion of dentinal tubules were investigated systematically in situ using human dentine slabs embedded in partial dentures. The dentine slabs were divided into three groups. In group I, the dentine slabs were kept in an oral cavity without plaque control. In group II, plaque was removed mechanically by brushing. Plaque control was carried out chemically using chlorhexidine in group III. After being kept in the oral cavities for 1, 2 and 3 weeks, the slabs were removed from the partial denture, followed by SEM observation to determine the morphological changes of the dentinal tubules. When no attempt was made to remove plaque, the diameter of tubule orifices increased to 390% of the original values within 3 weeks. In contrast, dentinal tubules were found to be occluded, i.e. the tubule orifices became <20% of the original value within 1 week when plaque control efforts were made, using either method of plaque control. We conclude that plaque control plays one of the key roles in the patency versus occlusion of dentinal tubules, and thus in the aetiology and natural reparative process of dentine hypersensitivity. PMID- 11380785 TI - Conduct of an algorithm in quantifying simulated palatal surface tooth erosion. AB - In order to test the ability of an algorithm to quantify simulated palatal erosion, a total of 10 extracted permanent upper central incisors were mounted in brass blocks. Baseline impressions were recorded using an addition cured silicone impression material in a metal impression tray. Once set and removed from the teeth, the impressions were coated twice with a high silver content electroconductive paint, applied using a brush, before being backed up with die stone to form an electroconductive replica. Each tooth was then subject to three treatments: application of phosphoric acid etchant gel for 60 s, application of etchant gel for 120 s and immersion for 3 h in Diet Coca-Cola*. After each one the replication process was repeated. Thereafter all replicas were mapped using a computer controlled electrical probe and the resultant digital terrain models (DTMS) compared using a surface matching and difference detection algorithm (SMADDA). Surface matching was unsuccessful only in one instance. As the duration of the insult increased, so did the proportion of the surface that underwent change to a maximum of 33.3%. Anatomical site was significantly (P < 0.05) associated with the susceptibility to erosion. The cingulum periphery appeared most resistant to this. The algorithmic approach offers much scope for monitoring dental erosion as acid dissolution of the tooth's surface appears to occur gradually. The cingulum region appears relatively more resistant to this process than other tooth sites and thus facilitates the process of surface matching. Further testing is however, required to determine precisely the algorithm's upper tolerance level. PMID- 11380786 TI - Colour stability of a new light-cured ceramic stain system subjected to glazing temperature. AB - A light-cured ceramic stain system (Orbit LC) offers promise as an effective method for surface characterization of ceramic restorations. This study evaluated colour change after glazing procedures for six commonly used stains from this novel system. One hundred and five ceramic specimens were made of the same enamel porcelain powder and standardized in size. The stains were applied on ceramic blocks and light-cured according to the manufacturer's recommendations. Porcelain colour was then measured with a calibrated colorimeter before and after each glazing procedure. The CIEL*a*b* colour co-ordinates were obtained as a measure of the direction and magnitude of colour change. Colour changes were noted for all six stains when subjected to the first glazing procedure (DeltaE* = 1.25 4.73). Brown, orange and green stains showed the greatest colour shifts (DeltaE* > 3.3), which might be considered clinically significant. However, the colour change is not of the same pattern in the colour space. Compared with the first glazing procedure, the second and third glazing procedures demonstrated less effect on the colour stability of the stains (DeltaE* = 0.36-1.12). The results provide important information about colour shifts in the system, which require overcorrection during stain application. PMID- 11380787 TI - Chewing ability as a parameter for evaluating the disability of patients with temporomandibular disorders. AB - Restoration of chewing ability is an important aspect of the treatment for temporomandibular disorders (TMDs). However, too little attention has been paid to it. We have used a questionnaire to evaluate and score the chewing ability of TMD patients. The questionnaire includes 19 kinds of food and a chewing task. The patient was asked if she/he experiences difficulty in enjoying eating. The aim of this study was to evaluate correlations between score of chewing ability (SCA) and other symptoms/signs of TMD. Four hundred and seventy-three consecutive TMD patients were evaluated for SCA and other symptoms/signs including temporomandibular joint (TMJ) pain, TMJ and muscle tenderness, TMJ noise (clicking and crepitus), and maximum mouth opening. The relationship between SCA and other symptoms/signs were analysed by multiple regression analysis. Score of chewing ability correlated significantly with TMJ pain and mouth opening capacity but not with TMJ noise and muscle tenderness. Age was a background factor but sex was not. The result of this study suggests that SCA correlated with dysfunction of the TMD patients. This method could be used to evaluate the ability of chewing in assessment of TMD. PMID- 11380788 TI - Quantitative description of temporomandibular joint sounds: defining clicking, popping, egg shell crackling and footsteps on gravel. AB - This study presents a quantitative description of temporomandibular joint (TMJ) sounds provided by a rule-based classification system based on sound classification by three dentists, who listened to and classified the sound recordings as no sound, click, coarse crepitus and fine crepitus. The sounds were recorded with microphones in the ear canal from 126 subjects during vertical opening, digitized at 15 000 Hz, and replayed using a computer sound card and speakers. The dentists' classification of a test set resulted in intra- and inter tester j values ranging from 0.71 to 0.81 and 0.61-0.73, respectively. Pooled j values for the dentists and the dentists plus the rules were 0.67 and 0.58, respectively, which were not significantly different in terms of the sound features on which the rules were based (P = 0.13). Linear discriminant analysis showed the four TMJ sound types were significantly different (P < 0.001). The performance of the rules was equivalent to the dentists and marginally better than the linear discriminant functions (P = 0.08), establishing the validity of the quantitative descriptions they provide. The recording and rebroadcast methodology produced sounds very similar to those observed in the clinic and could be used to train clinicians in classifying TMJ sounds. PMID- 11380789 TI - Alteration of the angle of the coronoid process in prognathism. AB - Muscles and bones may dynamically affect the functions of each other, resulting in changes in the activity of the muscle and/or morphological change in the bones. However, alterations of the morphology of the coronoid process have not been documented in relation to the temporal muscle between the mandible and the cranium. Angles of the coronoid process to the line through the right and the left frontotemporale were measured on posteroanterior (PA) cephalograms in patients with normal occlusions and Class II, division 1 malocclusions, and those with true skeletal Class III malocclusions. Standard deviation of the angle of the coronoid process in patients with Class III malocclusions was greater than those subjects with normal occlusion or Class II, division 1 malocclusions. Differences of angles between the right and the left coronoid process in each patient of either sex were significantly large in true skeletal Class III malocclusions (both P < 0.001). Large variation of angle of the coronoid process was related to prognathism, and this effect may represent adaptation of temporal muscle function to a variety of alterations in mandibular morphology. PMID- 11380790 TI - Quantitative study of bite force during sleep associated bruxism. AB - Nocturnal bite force during sleep associated bruxism was measured in 10 subjects. Hard acrylic dental appliances were fabricated for the upper and lower dentitions of each subject. Miniature strain-gauge transducers were mounted to the upper dental appliance at the right and left first molar regions. In addition, thin metal plates that contact the strain-gauge transducers were attached to the lower dental appliance. After a 1-week familiarization with the appliances, nocturnal bite force was measured for three nights at the home of each subject. From the 30 recordings, 499 bruxism events that met the definition criteria were selected. The above described system was also used to measure the maximum voluntary bite forces during the daytime. The mean amplitude of detected bruxism events was 22.5 kgf (s.d. 13.0 kgf) and the mean duration was 7.1 s (s.d. 5.3 s). The highest amplitude of nocturnal bite force in individual subjects was 42.3 kgf (15.6-81.2 kgf). Maximum voluntary bite force during the daytime was 79.0 kgf (51.8-99.7 kgf) and the mean ratio of nocturnal/daytime maximum bite force was 53.1% (17.3 111.6%). These data indicate that nocturnal bite force during bruxism can exceed the amplitude of maximum voluntary bite force during the daytime. PMID- 11380791 TI - Effect of cavity varnish, amalgam liner or dentin bonding agents on the marginal leakage of amalgam restorations. AB - The purpose of this in vitro study was to compare marginal leakage of Class II amalgam restorations lined with different agents (two new generation dentin bonding systems, a cavity varnish and an amalgam liner). Forty freshly extracted human molar teeth were divided into five groups. Class II preparations were prepared in the mesial and distal surfaces of each molar. While the cavities in two groups were lined with two new generation bonding systems, the other two groups were treated with an amalgam lining material and a cavity varnish. Fifth group was used as control, without any lining. Amalgam* was hand-condensed into each preparation. Specimens were thermocycled, stained and sectioned. Microleakage was graded using a stereomicroscope. Mean microleakage scores for occlusal and cervical margins were calculated and analysed. For the leakage values on occlusal margins, Kruskal-Wallis test indicated no significant difference in all groups. For the leakage values on cervical margins, Kruskal Wallis test indicated significant difference in four groups when compared with controls (P < 0.001). On the other hand, there was no significant difference between the occlusal and cervical leakage values in each group. PMID- 11380792 TI - Management of recurrent hepatitis C after liver transplantation. AB - Hepatitis C virus (HCV) reinfection is almost universal in patients transplanted for HCV-related cirrhosis. The medium-term survival after orthotopic liver transplantation (OLT) is similar to other transplanted patients, but the long term survival remains uncertain. The prevention and an effective treatment of progressive liver disease are the primary aims in HCV recurrence. Interferon and ribavirin, as monotherapy or in combination, have been tried to treat or prevent HCV recurrence. Preliminary studies suggest a better chance of initial HCV clearance and better results in preventing HCV recurrence with combination therapy. IFN or ribavirin, as monotherapy, may normalize liver enzymes, but only gives rise to a transient virological response, without histological improvement. Combination IFN and ribavirin may be able to prevent progression of HCV-related graft disease, but indications and duration of treatment need further evaluation. No clear association between type and dose of immunosuppressive and outcome of post-transplant HCV recurrence has been found. Strategies to minimize the effects of immunosuppressive drugs include dose reduction of all agents and the selective discontinuation of individual agents. Initial immunosuppression with a single drug may inhibit or delay the severe fibrosis, and further investigation with a single immunosuppressive regimen to evaluate the outcome of recurrent hepatitis C should be performed. The recent evidence that mycophenolate may have an antiviral effect needs a clinical confirmation. Retransplantation survival is better with early retransplantation, and for indications not directly related to viral recurrence. PMID- 11380793 TI - Identification of a cellular protein specifically interacting with the precursor of the hepatitis B e antigen. AB - In hepatitis B virus (HBV) the precore gene encodes a protein from which derives P22, the precursor of the mature secreted hepatitis B virus e antigen (HBeAg). Circumstantial evidences suggest that HBeAg and/or its precursor P22 are important for establishing persistent infection. Although P22 is essentially present in the secretory pathway, a substantial fraction has been found in the cytosol. In order to get new insights into the biological function of P22, we looked for cellular proteins which could strongly associate with this protein. Using immunoprecipitation studies on human cell extracts, we found that a non secreted cellular protein of about 32 kDa (P32) bound with a high specificity to P22. P32 associated neither with HBeAg nor with the viral core protein P21 which exhibits the same amino acids sequence as P22 but is N-terminally shorter by 10 residues. We also demonstrated that this interaction depended on the presence of the P22 C-terminal domain. Our data argues for a potential biological function of P22. PMID- 11380794 TI - Effect of HLA class II genotype on T helper lymphocyte responses and viral control in hepatitis C virus infection. AB - Hepatitis C virus (HCV) infection is very common worldwide, but has a broad range of outcomes. A minority of patients are able to clear infection spontaneously, and this is thought to be due to the emergence and maintenance of effective cell mediated immunity, particularly CD4+ T lymphocyte responses. Furthermore, genetic studies have indicated that HLA class II genotype strongly influences the outcome of infection. We have therefore investigated the influence of the protective HLA class II haplotype (DQB1*0301, which is in tight linkage disequilibrium with DRB1*1101) on the CD4+ T lymphocyte responses to HCV. We observe a strong association between this genotype and maintenance of a multispecific CD4+ T helper response. The effect on T helper responses was also maintained after combination interferon-alpha/ribavirin therapy, although the latter influenced the pattern of viral antigens to which patients responded. This is the first disease in which an association of HLA genotype with clinical outcome has been linked to an alteration of the immunological phenotype. The selection of protective peptides in those with the favourable HLA class II genotype may point in the direction of suitable vaccine candidates. PMID- 11380795 TI - Immunological response to interferon-gamma priming prior to interferon-alpha treatment in refractory chronic hepatitis C in relation to viral clearance. AB - The aim of this study was to clarify the immunological and virological responses to pre-administration of interferon-gamma prior to initiation of interferon-alpha treatment in patients with refractory chronic hepatitis C. Twenty-two nonresponders to 6-months of IFN-alpha treatment were enrolled. The hepatitis C virus (HCV) genotype was Ib in all. Natural IFN-gamma (1 MIU/day) was administered daily for 14 days followed by natural IFN-alpha (5 MIU/day) daily for 14 days and then three times weekly for 22 weeks. Serum immunological parameters (IL-10, neopterin, BMG, sCD8, sCD4, IL-6, IL-12) were measured as were the levels of several cytokines (IFN-gamma, TNF-alpha, IL-2, IL-4, IL-5, IL-6, IL 10). Three patients dropped out; two because of the occurrence of other diseases and one because of an adverse effect. At the end of the period of IFN-alpha treatment, HCV-RNA had become negative in six of 19 patients (end-of treatment response; ETR). Six months after the completion of IFN administration, a virological sustained response (SR) was seen in two of 19 patients. The mean serum levels of IL-10 were significantly decreased 6 weeks after the start of treatment. Other immunological parameter levels increased significantly during the period of IFN-gamma administration, and tended to return to the pretreatment level after the start of IFN-alpha administration. Univariate logistic regression analysis showed that the initial change in the levels of these parameters or the change in the ratios of Th1/Th2 parameter levels are useful factors indicative of the end of the treatment response. These findings suggest that priming with IFN gamma prior to the initiation of IFN-alpha treatment in patients with refractory chronic hepatitis C can modulate the host immune response and this might contribute to viral clearance. PMID- 11380796 TI - Possible role of human interleukin-6 and soluble interleukin-6 receptor in hepatitis B virus infection. AB - Human interleukin-6 has been shown to promote hepatitis B virus (HBV) infection. However, it is not clear whether this influence is the result of a direct interaction between interleukin-6 (IL-6) and the HBV envelope proteins or of a rather indirect mechanism. A direct interaction of IL-6 and the preS region of the large envelope protein (L-protein) of HBV has been reported. In this study we assessed the binding of IL-6 and of the IL-6 receptor subunits to the preS region of the L-protein of HBV. Binding of IL-6 and IL-6 receptor subunits sIL-6R and gp130 to preS was assessed by immunoprecipitation with recombinant preS proteins. In patient sera IL-6 and sIL-6R concentrations were analysed with respect to the course of hepatitis B infection during and after interferon-alpha (IFN-alpha) therapy. The IL-6 and IL-6 receptor subunits could not be precipitated with recombinant preS proteins. In sera of patients who responded to IFN-alpha therapy by virus elimination, a significant increase in sIL-6R concentration was measured. No increase in sIL-6R levels was seen in patients who did not respond to IFN-alpha. Hence, IL-6 and IL-6 receptor subunits do not bind to preS directly. A possible role for sIL-6R in the elimination of HBV infection is discussed. PMID- 11380797 TI - In vitro effect of thymosin-alpha1 and interferon-alpha on Th1 and Th2 cytokine synthesis in patients with chronic hepatitis C. AB - Current evidence suggests that increased expression of Th1-associated cytokines is important for immune-mediated eradication of hepatitis C infection, while an increase in Th2-associated cytokines is associated with persistence of infection. In this study we evaluated the effects of thymosin-alpha1 (TA1), a naturally occurring thymic peptide, and interferon-alpha (IFN-alpha) on cytokine production in peripheral blood mononuclear cells from untreated patients with chronic hepatitis C. We examined the effect of incubation with TA1, IFN-alpha, or both, on production of Th1-associated cytokines (IL-2, IFN-gamma), Th2-associated cytokines (IL-4, IL-10), and synthesis of the antiviral protein 2',5' oligoadenylate synthetase. TA1 treatment induced a significant increase in production of IL-2 and 2',5'-oligoadenylate synthetase. Smaller increases were also seen after treatment with IFN-alpha, while incubation with TA1 and IFN-alpha together led to an additive or synergistic effect. Incubation with TA1 resulted in a decrease in IL-4 and IL-10, whereas IFN-alpha increased these cytokines. The addition of TA1 to IFN-alpha significantly reversed this IFN-alpha-induced increase. Hence, TA1 treatment could benefit patients with hepatitis C infection by increasing the Th1-type response, fundamental for sustained clearance of hepatitis C; and by decreasing the Th2-type response, associated with persistence of viraemia. PMID- 11380798 TI - Pilot study of interferon-alpha and ribavirin treatment in patients with chronic hepatitis C and normal transaminase values. AB - Combination interferon-alpha (IFN-alpha) and ribavirin treatment has become standard therapy for patients with chronic hepatitis C and elevated transaminase levels (> 1.5 x upper limit of normal). No previous study has specifically examined the efficacy of this treatment in patients with normal transaminase values. In this pilot study, we treated 19 patients, with normal or near-normal ALT values on at least three occasions, and histologically mild disease, with induction IFN-alpha2b, 5 mega units daily for 4 weeks, then 3 mega units thrice weekly for 44 week, plus concomitant ribavirin 1000 mg or 1200 mg daily for 48 weeks. Nine of the 19 (47%) showed sustained virological response, defined as undetectable HCV-RNA at 24 weeks after the end of treatment. No ALT flares were observed in any patient. We conclude that combination induction IFN-alpha and ribavirin therapy may be effective in patients with normal-ALT, and appears not to induce flares of ALT activity. Controlled trials of this treatment in this subgroup of patients with hepatitis C are warranted. PMID- 11380799 TI - Hepatitis C virus genotypes: distribution and clinical significance in patients with cirrhosis type C seen at tertiary referral centres in Europe. AB - The aim of this study was to evaluate the distribution and clinical significance of hepatitis C virus (HCV) genotypes in European patients with compensated cirrhosis due to hepatitis C (Child class A) seen at tertiary referral centres. HCV genotypes were determined by genotype-specific primer PCR in 255 stored serum samples obtained from cirrhotics followed for a median period of 7 years. Inclusion criteria were biopsy-proven cirrhosis, absence of complications of cirrhosis and exclusion of all other potential causes of chronic liver disease. The proportion of patients with types 1b, 2, 3a, 1a, 4 and 5 were 69%, 19%, 6%, 5%, 0.5% and 0.5%, respectively. Kaplan-Meier 5-year risk of hepatocellular carcinoma (HCC) was 6% and 4% for patients infected by type 1b and non-1b, respectively (P=0.8); the corresponding figures for decompensation were 18% and 7% (P=0.0009) and for event-free survival were 79% and 89% (P=0.09), respectively. After adjustment for baseline clinical and serological features, HCV type 1b did not increase the risk for HCC [adjusted relative risk=1.0 (95% confidence interval=0.47-2.34)], whereas it increased the risk for decompensation by a factor of 3 (1.2-7.4) and decreased event-free survival by a factor of 1.7 (0.9-3.10). In conclusion, type 1b and, to a lesser extent, type 2, are the most common HCV genotypes in European patients with cirrhosis. HCV type 1b is not associated with a greater risk for HCC, but increases the risk for decompensation by threefold in patients with cirrhosis. PMID- 11380800 TI - Quantitative evaluation of cytomegalovirus DNA in infantile hepatitis. AB - We used a PCR method to develop a diagnostic assay for the detection of cytomegalovirus (CMV) DNA in infantile hepatitis, which has been suggested to be associated with CMV infection. CMV DNA was detected in 25 (58.1%) of 43 patients with elevated serum alanine aminotransferase (ALT) levels but no jaundice, and no hepatitis B or C as assessed by conventional PCR. None of the samples from 97 healthy infants tested positive for CMV DNA. We assayed CMV DNA quantitatively in blood using a real-time PCR system that allowed reproducible detection of at least 10 copies of CMV DNA. When 1 microg of DNA from each blood sample was used in this system, a good correlation was obtained between the calculated and measured copy numbers of CMV DNA. This system detected CMV DNA in 29 patients (67.4%) with liver dysfunction. Serial studies in patients with liver dysfunction revealed that CMV DNA copy number decreased, ultimately to below 10, as the ALT levels normalized. In contrast, no CMV DNA copies were detectable by the real time system in any of the samples from control subjects. These results highlight the usefulness of detecting CMV DNA in the diagnosis of infantile hepatitis and indicate that the real-time quantitative PCR assay may be a valuable tool for monitoring CMV-associated infantile hepatitis. PMID- 11380801 TI - Prevalence of anti-hepatitis E virus antibodies in different Indian animal species. AB - Prevalence of IgG antibodies to hepatitis E virus (IgG-anti-HEV) was determined among different animal species from India. Seropositivity varied from 4.4% to 6.9% in cattle, 54.6-74.4% in pigs and 2.1-21.5% in rodents. Of the 44 dogs screened, 10 were positive (22.7%). None of the 250 goat sera tested were found to be anti-HEV positive. Among rodents, over 50% serum samples collected in 1985 from Bandicota bengalensis were positive for anti-HEV antibodies. No evidence of HEV infection was obtained following experimental inoculation of an Indian strain (AKL-90) of HEV into anti-HEV negative pigs and goats. The results document varied prevalence of anti-HEV antibodies in different animal species from India and of inability of Indian pigs and goats to support replication of at least one human strain of HEV. PMID- 11380802 TI - Failure to infect rhesus monkeys with hepatitis C virus strains of genotypes 1a, 2a or 3a. AB - The chimpanzee is the only recognized animal model for the study of hepatitis C virus (HCV). However, recently it was reported that rhesus monkeys were susceptible to HCV and developed hepatitis during infection. In the present study, we inoculated two rhesus monkeys each with HCV strain H77 (genotype 1a), strain HC-J6 (genotype 2a) or strain S52 (genotype 3a). Weekly serum samples were tested for liver enzyme values, HCV antibodies and HCV RNA. We did not find evidence of HCV infection in any of the monkeys during 24 weeks of follow-up. Our study demonstrates that rhesus monkeys are not readily infected with HCV and apparently do not represent a useful animal model for the study of HCV. PMID- 11380803 TI - Treatment prospects for autosomal-dominant polycystic kidney disease. AB - An increased understanding of the molecular genetic and cellular pathophysiologic mechanisms responsible for the development of autosomal-dominant polycystic kidney disease (ADPKD), made possible by the advances in molecular biology and genetics of the last three decades, has laid the foundation for the development of effective therapies. As the concept that a polycystic kidney is a neoplasm in disguise is becoming increasingly accepted, the development of therapies for ADPKD may benefit greatly from the expanding body of information on cancer chemoprevention and chemosuppression. This review summarizes the observations that already have been made and discusses therapies for PKD that deserve investigation. PMID- 11380804 TI - Hepatocyte growth factor: renotropic role and potential therapeutics for renal diseases. AB - Hepatocyte growth factor (HGF), a ligand for the c-Met receptor tyrosine kinase, has mitogenic, motogenic, anti-apoptotic, and morphogenic (for example, induction of branching tubulogenesis) activities for renal tubular cells, while it has angiogenic and angioprotective actions for endothelial cells. Stromal cells such as mesangial cells, endothelial cells, and macrophages are sources of renal HGF; thus, HGF mediates epithelial-stromal and endothelial-mesangial interactions in the kidney. In response to acute renal injury, the expression of HGF increases in the injured kidney and in distant intact organs such as the lung and spleen. Locally and systemically increased HGF supports renal regeneration, possibly not only by enhancing cell growth but also by promoting morphogenesis of renal tissue. During progression of chronic renal failure/renal fibrosis, the expression of HGF decreases in a manner reciprocal to the increase in expression of transforming growth factor-beta (TGF-beta), a key player in tissue fibrosis. A decrease in endogenous HGF, as well as increase in TGF-beta, augments susceptibility to the onset of chronic renal failure/renal fibrosis. On the other hand, supplements of exogenous HGF have preventive and therapeutic effects in cases of acute and chronic renal failure/renal fibrosis in laboratory animals. HGF prevents epithelial cell death and enhances regeneration and remodeling of renal tissue with injury or fibrosis. A renotropic system underlies the vital potential of the kidney to regenerate, while an impaired renotropic system may confer susceptibility to the onset of renal diseases. Thus, HGF supplementation may be one therapeutic strategy to treat subjects with renal diseases, as it enhances the intrinsic ability of the kidney to regenerate. PMID- 11380805 TI - Arachidonic acid directly activates members of the mitogen-activated protein kinase superfamily in rabbit proximal tubule cells. AB - BACKGROUND: To explore the roles of eicosanoids in arachidonic acid-induced mitogen-activated protein kinase (MAPK) signal transduction, we have shown that exposure of proximal tubular cells to arachidonic acid induces phosphorylation of c-Jun NH2-terminal kinase (JNK) and extracellular signal-regulated kinase (ERK), two members of the MAPK superfamily. We observed that ketoconazole, an inhibitor of the cytochrome P450 pathway, blocked ERK but not JNK activation. METHODS: Direct regulation of arachidonic acid on mitogen-activated protein kinase (MAPK) signaling pathways was evaluated more directly by utilizing specific enzyme inhibitors of the cytochrome P450 metabolic pathway and by comparing the relative efficacy of arachidonic acid versus its cytochrome P450 metabolites (exogenous and endogenous), eicosatetraynoic acid (ETYA), and other fatty acids on the phosphorylation of members of the MAPK superfamily (ERKs, JNK, and p38(MAPK)), by utilizing early passage rabbit proximal tubular epithelial cells. RESULTS: Arachidonic acid activated p38(MAPK), a third member of the MAPK superfamily, in a time- and concentration-dependent manner. Studies designed to evaluate the ability of arachidonic acid and its cytochrome P450 metabolites (endogenously and exogenously) to stimulate ERKs, JNK, and p38(MAPK) found four conclusions. First, the metabolites of arachidonic acid generated endogenously by cytochrome P450 2C1 significantly augmented basal ERK activity, whereas the metabolites generated by the 2C2 isozyme significantly augmented basal p38(MAPK) activity. However, their effects were less profound than arachidonic acid itself. In contrast, there were no significant effects with transfection of either isozyme on basal JNK activity. Second, a variety of exogenous cytochrome P450 products were less potent than arachidonic acid on a molar basis in stimulating the activity of all three MAPKs. Third, ketoconazole and 17-octadecynoic acid, inhibitors of the cytochrome P450 pathway, as well as PPOH and DDMS, inhibitors of the epoxygenase and omega hydroxylase pathways, respectively, failed to significantly reduce the effects of arachidonic acid to activate ERK and p38(MAPK) (JNK was not evaluated). Finally, arachidonic acid, its inactive analog ETYA, and other fatty acids with differing chain lengths and degrees of saturation stimulated the activity of all three MAPKs. CONCLUSIONS: These observations substantiate a role for arachidonic acid and other fatty acids in signaling linked to the MAPK superfamily in rabbit proximal tubular epithelium without the necessity of conversion to cytochrome P450 metabolites. PMID- 11380806 TI - Release of urokinase plasminogen activator receptor during urosepsis and endotoxemia. AB - BACKGROUND: The urokinase receptor (uPAR; CD87) is a multifunctional molecule involved in fibrinolysis, in proteolysis, in renal tubular functions, and in migration and adhesion of inflammatory cells to the site of infection. METHODS: To gain insight into systemic and local release of uPAR and into its regulation during urosepsis, which is one of the leading causes of chronic renal failure, uPAR was measured in urine and plasma of healthy human controls (N = 20), patients with culture-proven urosepsis (N = 30), and healthy human volunteers intravenously injected with endotoxin (N = 7). RESULTS: Patients had elevated uPAR levels in both plasma and urine. Three hours after endotoxin challenge in volunteers, there was also a significant increase of uPAR in plasma and in urine. The urine/plasma ratio for uPAR was highly elevated during urosepsis and experimental endotoxemia, suggesting local production in the kidney. Accordingly, damaged tubuli strongly expressed uPAR during pyelonephritis. Moreover, tubular epithelial cells produced uPAR in vitro, and this secretion was strongly up regulated after stimulation with interleukin-1 beta or tumor necrosis factor alpha. CONCLUSIONS: We found that uPAR is released systemically and in the urinary tract during urosepsis and experimental endotoxemia. This systemic and renal production of uPAR during pyelonephritis may play a central role in eliminating the infection and protecting renal function. PMID- 11380807 TI - Lipoprotein-stimulated mesangial cell proliferation and gene expression are regulated by lipoprotein lipase. AB - BACKGROUND: Hyperlipidemia accelerates the progression of glomerular disease, and lipoproteins bind glomerular mesangial cells (MC) and induce proliferation and cytokine expression. In the vessel wall, the binding of lipoproteins to endothelial cells is markedly enhanced by lipoprotein lipase (LpL), synthesized by the underlying smooth muscle cells. While it is known that LpL is localized to the glomerulus, it is not known if and how it modulates the lipoprotein-mesangial interaction. METHODS: Very low-density lipoprotein (VLDL) was isolated from rats and was used to treat cultured primary rat MCs. Binding studies were done with and without LpL and with/without pretreatment with heparanase, which degrades cell surface heparan sulfate proteoglycan (HSPG), known to modulate the LpL lipoprotein interaction in blood vessels. VLDL/LpL was also used to assess MC proliferation and gene expression of the cytokine platelet-derived growth factor (PDGF). RESULTS: LpL enhanced VLDL binding to MCs by as much as 200-fold, and most of this effect was blocked by pretreatment with heparanase. LpL amplified VLDL-driven MC proliferation and increased VLDL-induced PDGF expression. Heparanase pretreatment of cells eliminated both of these amplifications. LpL alone increased MC proliferation and PDGF gene expression. DISCUSSION: As in the vessel wall, LpL enhances VLDL binding to MCs. MCs respond to LpL binding by proliferating and expressing cytokines such as PDGF. LpL may be a crucial paracrine mediator of the glomerular response to circulating lipoproteins, amplifying a response that includes cytokine elaboration, influx of circulating monocytes, and eventual sclerosis. PMID- 11380808 TI - Red cell traverse through thin glomerular basement membranes. AB - BACKGROUND: How red cells enter the urinary filtrate in most cases of hematuria of glomerular origin has remained a mystery despite the frequent ultrastructural examination of renal biopsy material. METHODS: Serial sections of glutaraldehyde fixed, resin-embedded material from a case of sporadic microhematuria were examined by transmission electron microscopy when the site of a red cell traversing the glomerular capillary wall was fortuitously discovered on routine examination. RESULTS: The red cell assumed a dumbbell shape and traversed a localized gap 2.25 microm in diameter in the glomerular endothelium and basement membrane. Serial sections suggested a transcellular route. Apart from the thinning of the basement membrane (167 nm), there were no other generalized abnormalities. CONCLUSION: Red cells can traverse through gaps in the glomerular capillary walls to gain access to Bowman's space. This may be the origin of glomerular hematuria in common noninflammatory forms of glomerular disease, including thin basement membrane nephropathy. PMID- 11380809 TI - Calpain inhibitor-1 reduces renal ischemia/reperfusion injury in the rat. AB - BACKGROUND: Activation of the cysteine protease calpain has been implicated in renal ischemia/reperfusion (I/R) injury. The aim of this study was to investigate the effects of calpain inhibitor-1 (Cal I-1) in an in vivo model of renal I/R injury. METHODS: Male Wistar rats were administered Cal I-1 (10 mg/kg, IP) 30 minutes before undergoing bilateral renal ischemia (45 minutes) followed by reperfusion (6 hours). Plasma concentrations of urea, creatinine, Na(+), gamma glutamyl transferase (gamma GT), aspartate aminotransferase (AST) and urinary Na(+), glutathione S-transferase (GST), and N-acetyl-beta-D-glucosaminidase (NAG) were measured for the assessment of renal dysfunction and I/R injury. Creatinine clearance (C(Cr)) and fractional excretion of Na(+) (FE(Na)) were used as indicators of glomerular and tubular function, respectively. Kidney myeloperoxidase (MPO) activity and malondialdehyde (MDA) levels were measured for assessment of neutrophil infiltration and lipid peroxidation, respectively. Renal sections were used for histologic grading of renal injury and for immunohistochemical localization of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). RESULTS: Cal I-1 significantly reduced I/R-mediated increases in urea, creatinine, gamma GT, AST, NAG, and FE(Na) and significantly improved C(Cr). Cal I-1 also significantly reduced kidney MPO activity and MDA levels. Cal I-1 also reduced histologic evidence of I/R-mediated renal damage and caused a substantial reduction in the expression of iNOS and COX-2, both of which involve activation of nuclear factor-kappa B (NF-kappa B). CONCLUSIONS: : These results suggest that Cal I-1 reduces the renal dysfunction and injury associated with I/R of the kidney. We suggest that the mechanism could involve the inhibition of I/R-mediated activation of NF-kappa B. PMID- 11380810 TI - Cell surface heparan sulfate proteoglycans control the response of renal interstitial fibroblasts to fibroblast growth factor-2. AB - BACKGROUND: While the progression of renal disease to end stage is strongly correlated with tubulointerstitial changes, the control of the fibrotic process within the interstitium is poorly understood. Basic fibroblast growth factor (FGF 2) has been implicated as a major growth factor involved in fibroblast activation and extracellular matrix synthesis. Furthermore, in many cells, the activity of FGF-2 is controlled by a low-affinity but high-capacity interaction with heparan sulfate (HS) proteoglycans (PGs), such as members of the syndecan family. These molecules are likely to be central to the control of interstitial fibrosis, but as yet, there has been no characterization of their synthesis by interstitial cells. METHODS: The expression of HSPG on the surface of NRK 49F fibroblasts was demonstrated by immunohistochemistry and by metabolic labeling with [(35)S] sulfate. HSs were characterized by specific enzymatic digestion, size exclusion chromatography, and anion exchange chromatography. The mRNA for syndecan 1 through syndecan 4 in the fibroblasts was detected by semiquantitative reverse transcription-polymerase chain reaction. Fibroblast proliferation was measured by the MTT assay. RESULTS: Immunohistochemistry and [(35)S]-sulfate-labeling demonstrated that renal fibroblasts expressed HSPGs on their surface. Furthermore, enzymatic removal of these HS (but not chondroitin sulfate) glycosaminoglycan (GAG) chains, or inhibition of GAG sulfation, abolished the proliferative response of both NRK cells and primary human cortical fibroblasts to FGF-2 but not to platelet-derived growth factor. The addition of conditioned medium, containing HS-GAG fragments, restored the proliferative response to FGF 2, confirming the specificity of the interaction. Finally, the mRNA for all four syndecans was detected in the fibroblasts, and that for syndecan 1 in particular was up-regulated by FGF-2. CONCLUSIONS: The present study demonstrates that the expression of cell surface HSPG was essential for the proliferation of renal fibroblasts in response to FGF-2, and therefore may play a major role in the development and persistence of a proliferating phenotype during interstitial nephritis. PMID- 11380811 TI - Fluvastatin suppresses oxidative stress and fibrosis in the interstitium of mouse kidneys with unilateral ureteral obstruction. AB - BACKGROUND: Recently, we demonstrated increased oxidative stress in the interstitium of ureteral obstructed kidneys based on the increased expression of heme oxygenase-1 and immunohistochemical detection of advanced glycation end products (AGE) in the interstitium. Antioxidant therapy may have a therapeutic potential toward interstitial fibrosis of unilateral ureteral obstruction (UUO) kidneys. Fluvastatin is an HMG-CoA reductase inhibitor and has been demonstrated to have an antioxidant activity in vitro. METHODS: The effects of fluvastatin on UUO kidneys from the viewpoints of antioxidant action in vivo and antifibrosis action were studied. To investigate the antioxidant action and its therapeutic efficacy of fluvastatin in UUO kidneys, AGE accumulation and fibrosis in the obstructed kidneys was compared among vehicle-, pravastatin-, or fluvastatin treated (10 or 40 mg/kg/day) groups. RESULTS: Tubulointerstitial fibrosis was significantly attenuated in fluvastatin-treated animals. Fluvastatin significantly suppressed the degree of immunostaining of AGE in UUO kidneys. CONCLUSIONS: These results provide evidence for the antioxidant action of fluvastatin in vivo. The decreased interstitial fibrosis along with a decreased oxidative stress marker in the interstitial lesion strongly suggests the existence of a causal relationship between them. Fluvastatin may have therapeutic value in slowing or preventing interstitial fibrosis in progressive renal disease. PMID- 11380812 TI - Glomerular cell number in normal subjects and in type 1 diabetic patients. AB - BACKGROUND: The number of cells in glomeruli has been a challenging measure, especially in human kidneys, with only a small amount of tissue obtained by biopsy. However, the number of cells and their function are important determinants of renal function in health and disease. METHODS: Modern morphometric techniques have now provided the means to determine the numerical density (Nv) and number (with a measure of glomerular volume) of endothelial cells, mesangial cells, and podocytes in plastic-embedded renal tissue biopsied from nondiabetic subjects (N = 36) and type 1 diabetic patients (N = 46) over an extended age range from childhood through late adult. RESULTS: Nv values for all glomerular cells varied only slightly with age and did not change within the range of glomerular lesions of diabetes studied. Thus, the increase in glomerular volume during childhood to a steady level thereafter was the primary determinant of total glomerular cell number. The number of mesangial cells and endothelial cells increased with age, reflecting the increase in all cells, while the podocytes remained unchanged in number over all ages studied (10 to 69 years). Numbers of total glomerular cells, mesangial cells, and endothelial cells were not changed with diabetes, while podocytes were fewer in number in diabetic patients of all ages, with reduced podocyte numbers even in diabetes of short duration. CONCLUSIONS: The essentially constant glomerular cell density in nondiabetic and diabetic subjects under different circumstances possibly indicates an underlying propensity for the glomerulus to regulate its architecture to maintain a constant number of cells per volume, no matter the size of the glomerulus or the severity of diabetic nephropathy studied in this set of patients. The reductions in podocyte numbers in both younger and older diabetic patients indicate a significant risk for functional abnormalities as diabetic nephropathy progresses. Moreover, these observations do not support the suggestion of marked increases in glomerular cell number (and especially mesangial cells) with the development and progression of diabetic nephropathy. PMID- 11380813 TI - Enhanced protection from renal ischemia-reperfusion [correction of ischemia:reperfusion] injury with A(2A)-adenosine receptor activation and PDE 4 inhibition. AB - BACKGROUND: We previously demonstrated in rats and mice that agonists of A(2A) adenosine receptors (A(2A)-ARs) reduce renal injury following ischemia reperfusion. We now extend these studies and examine the effects of ATL-146e (formerly DWH-146e), an A(2A)-AR agonist, and rolipram, a type IV phosphodiesterase (PDE 4) inhibitor, on murine renal injury following ischemia reperfusion. METHODS: C57BL/6 mice were treated with rolipram, ATL-146e, or both compounds combined and were subjected to renal ischemia for 32 minutes and reperfusion for 24 to 48 hours. In vitro studies were performed on suspended and adhering human neutrophils. RESULTS: Continuous delivery of rolipram or ATL-146e during reperfusion reduced renal injury in a dose-dependent manner. Maximal protection was observed when ATL-146e was infused for six hours during reperfusion. Elevated plasma creatinine and myeloperoxidase activity produced by ischemia-reperfusion were reduced by rolipram (0.1 ng/kg/min) and ATL-146e (10 ng/kg/min) by up to approximately 60% and 70%, respectively. Co-infusion of both compounds produced a maximum reduction of plasma creatinine of approximately 90% and myeloperoxidase activity. In vitro studies on suspended and adhering human neutrophils demonstrated that selective stimulation of A(2A)-ARs by ATL-146e increased cAMP accumulation, reduced oxidative activity of activated neutrophils, and decreased activated neutrophil adherence. These responses were potentiated by rolipram. CONCLUSIONS: We conclude that the combined infusion of ATL-146e and rolipram leads to enhanced renal tissue protection from ischemia-reperfusion by mechanisms that may include reduced neutrophil adherence/recruitment and release of reactive oxygen species. PMID- 11380814 TI - Association of parvovirus B19 infection with idiopathic collapsing glomerulopathy. AB - BACKGROUND: Collapsing glomerulopathy (CG), a disorder with severe glomerular and tubular involvement, occurs either as an idiopathic lesion or in some patients with human immunodeficiency virus (HIV) infection known as HIV-associated nephropathy (HIVAN). We previously reported a renal transplant recipient with de novo CG and red cell aplasia in association with persistent parvovirus B19 (PVB19) infection. This prompted us to look for an association between PVB19 infection and CG. METHODS: DNA from archived biopsies of patients with CG was analyzed for PVB19 by polymerase chain reaction (PCR). Results were compared with HIVAN, idiopathic focal segmental glomerulosclerosis (FSGS), and controls. In situ hybridization (ISH) was done to localize PVB19 in renal biopsies. Peripheral blood specimens of patients with CG, HIV infection, healthy controls, and randomly selected hospitalized patients (sick controls) were also analyzed for PVB19. RESULTS: PVB19 DNA was detected in renal biopsies of 18 out of 23 (78.3%) patients with CG, 3 out of 19 (15.8%) with HIVAN, 6 out of 27 (22.2%) with FSGS, and 7 out of 27 (25.9%) controls (P < 0.01, CG vs. HIVAN, FSGS, and controls). PVB19 was detected in peripheral blood of 7 out of 8 (87.5%) CG patients, 3 out of 22 (13.6%) with HIV infection, 4 out of 133 (3%) healthy controls, and 2 out of 50 (4%) sick controls (P < 0.001, CG vs. HIV infected, healthy, and sick controls). PVB19 was identified in glomerular parietal and visceral epithelial and tubular cells by ISH. CONCLUSIONS: The significantly higher prevalence of PVB19 DNA in renal biopsies and peripheral blood of CG patients suggests a specific association between PVB19 infection and CG. In susceptible individuals, renal epithelial cell infection with PVB19 may induce CG. PMID- 11380815 TI - Electroporation-mediated PDGF receptor-IgG chimera gene transfer ameliorates experimental glomerulonephritis. AB - BACKGROUND: Mesangial cell proliferation and phenotypic alteration occur in an early phase of glomerular injury and precede increased extracellular matrix accumulation. A critical growth factor responsible for mesangial proliferation is platelet-derived growth factor (PDGF), which has proved to be a potent mitogen. METHODS: We generated a chimeric cDNA encoding an extracellular domain of the beta-PDGF receptor fused with IgG-Fc, termed PDGFR/Fc, and examined the feasibility of gene therapy targeting PDGF using PDGFR/Fc. RESULTS: Chimeric PDGFR/Fc molecule completely inhibited the tyrosine phosphorylation of beta-PDGF receptors and cellular proliferation induced by PDGF in vitro. We then introduced the PDGFR/Fc expression vector into the muscle of anti-Thy-1 model of glomerulonephritic rats by electroporation. The plasma concentration of chimeric PDGFR/Fc levels was 244.4 +/- 89.8 ng/mL four days after transfection. On day 5, PDGFR/Fc gene transfer significantly reduced the number of PCNA-positive cells and glomerular cell numbers by 59.6 and 23.2%, respectively. Northern blot analysis demonstrated that glomerular mRNA levels of alpha-smooth muscle action, transforming growth factor-beta 1, and type I collagen were also suppressed on days 5 and 7 by the PDGFR/Fc transfection. There was a significant reduction in the matrix score of the transfected nephritic rats (2.91 +/- 0.75 and 2.06 +/- 0.95; disease control group vs. treated group, P < 0.001). CONCLUSION: These results suggest that gene therapy by the manipulation of PDGF action using electroporation-mediated PDGFR/Fc gene transfer to the skeletal muscle might be a useful treatment for mesangioproliferative glomerulonephritis. PMID- 11380816 TI - t-PA promotes glomerular plasmin generation and matrix degradation in experimental glomerulonephritis. AB - BACKGROUND: In addition to its well-known role in degrading fibrin, recent evidence suggests that plasmin degrades matrix proteins and activates prometalloproteinases. Plasmin is generated from plasminogen by tissue plasminogen activator (t-PA). We hypothesized that t-PA treatment increases plasmin generation in nephritic glomeruli and degrades pathological matrix leading to a therapeutic reduction in matrix accumulation. METHODS: Anti-Thy-1 nephritis was induced by injection of OX-7 antibody. Rats were given twice daily intravenous injections of saline (disease control group) or human recombinant t PA (rt-PA; 1 mg/kg body weight) on days 3 through 5. Proteinuria, glomerular matrix protein staining, and glomerular mRNA levels for transforming growth factor-beta 1 (TGF-beta 1), fibronectin, and plasminogen activator inhibitor type 1 (PAI-1) were evaluated at day 6. Localization of rt-PA, plasmin generation by glomeruli in vitro, and glomerular production and content of active TGF-beta1 were also investigated. RESULTS: Compared with disease control animals, proteinuria and staining score for periodic acid-Schiff (2.75 +/- 0.17 vs. 1.41 +/- 0.09), fibronectin-EDA+ (19 +/- 2 vs. 14 +/- 1), laminin (35 +/- 2 vs. 25 +/- 2), type I collagen (33 +/- 1 vs. 21 +/- 3), and type IV collagen (27 +/- 2 vs. 23 +/- 1) were significantly reduced in treated rats (P < 0.01). Glomerular TGF beta 1, fibronectin, and PAI-1 mRNA levels were unchanged. rt-PA colocalized with fibrin along glomerular capillary walls and in the mesangium. Nephritic glomeruli in vitro had decreased plasmin activity, which was elevated by an in vivo presacrifice injection of rt-PA. Glomerular production and content of active TGF beta 1 were unchanged by the rt-PA injection. CONCLUSIONS: : These results show that injected rt-PA binds to fibrin in nephritic glomeruli, thus increasing plasmin generation and promoting pathological matrix degradation without activating latent TGF-beta. Agents that increase plasmin generation, such as t PA, may have potential as antifibrotic therapies. PMID- 11380817 TI - Significance of histologic patterns of glomerular injury upon long-term prognosis in severe lupus glomerulonephritis. AB - BACKGROUND: Patients with systemic lupus erythematosus have a spectrum of glomerular disease, but the different patterns of glomerular injury identified within the general category of "severe" lupus glomerulonephritis are responsible for much of the morbidity and mortality in this disease. The glomerular injury patterns seen with severe lupus glomerulonephritis have been separated into distinct histopathologic groups to determine whether they can predict long-term patient outcome. METHODS: We analyzed the clinical follow-up of 85 patients participating in a controlled prospective therapeutic trial for the treatment of severe lupus glomerulonephritis conducted from April 1981 to December 1988, with an average follow-up of 10 years. Patients were classified according to the 1982 World Health Organization classification for lupus glomerulonephritis. RESULTS: During the course of follow-up [120 +/- 65 (SD) months], 60% of patients with category IV (diffuse proliferative glomerulonephritis) lesions entered a remission compared with only 38% of patients with category III (> or =50%, focal and segmental glomerulonephritis) lesions and 27% of patients with category Vc (> or =50%) and Vd (P < 0.05). Renal survival at 10 years was 75% for those with category IV lesions, 47% for patients with category Vc (> or =50%) and Vd, and 52% for patients with category III (> or =50%) lesions (P < 0.05). Based on multivariate analysis, patients with category III (> or =50%) or Vc (> or =50%) and Vd lesions had a relative risk of progression to end-stage renal disease 2.9 times that of category IV patients (P < 0.01), while the likelihood of entering a remission was 8.2 times greater for category IV patients (P = 0.0001). CONCLUSION: The histopathologic categorization among patients with severe lupus glomerulonephritis provides information relevant to their long-term outcome. PMID- 11380818 TI - Chronic aristolochic acid toxicity in rabbits: a model of Chinese herbs nephropathy? AB - BACKGROUND: Chinese herbs nephropathy (CHN) is a new type of subacute interstitial nephritis that is attributed to aristolochic acid (AA), which inadvertently has been included in slimming pills. The contribution of other simultaneously prescribed drugs remains disputed. In the present study, the effects of a chronic intake of AA given as a single drug was evaluated through renal histology and function in rabbits. METHODS: Female New Zealand White rabbits were injected intraperitoneally with either 0.1 mg AA/kg or with saline 5 days a week for 17 to 21 months. Body weight, renal function, and urinary excretion of glucose and low molecular weight proteins were monitored prior to sacrifice at the end of the study period. RESULTS: All animals given AA developed renal hypocellular interstitial fibrosis, which was classified into three patterns. Fibrosis was confined to medullary rays (MRs) in pattern I (N = 3), extended to the outer cortical labyrinth (OCL) in pattern II (N = 2), and eventually to the inner cortical labyrinth (ICL) in pattern III (N = 6). Fibrosis in MR and OCL was associated with mainly proximal tubular epithelial cell flattening. All treated animals displayed urothelial atypia. Three of them also developed tumors of the urinary tract. No significant pathologic changes were found in control rabbits. AA-treated animals differed from controls by an impaired growth, increased serum creatinine, glucosuria, tubular proteinuria, and anemia. CONCLUSION: The observed pattern of renal histopathological lesions and disorders of the renal function, as well as urothelial atypia and malignancy, are very reminiscent of CHN. Our observations therefore support a causal role of AA alone in the genesis of this new nephropathy. PMID- 11380819 TI - Nuclear factor-kappa B binding to the HIV-1 LTR in kidney: implications for HIV associated nephropathy. AB - BACKGROUND: We have recently shown that renal epithelium is infected by HIV-1 and supports HIV-1 transcription in seropositive patients with renal disease. To investigate the regulation of HIV-1 gene expression in kidney, an HIV-1 transgenic mouse model was used to analyze the host transcriptional proteins that bind the 5' long-terminal repeat (LTR). METHODS: Viral gene expression was assessed in transgenic mouse tissue using Northern blotting and mRNA in situ hybridization. The transcription factors involved in LTR binding were determined using electrophoretic mobility shift assays. Cytoplasmic and nuclear extracts were prepared from tissues with varied levels of transgene expression. The binding of transcription factors to specific LTR fragments was determined using DNA competition experiments and supershifts with transcription factor-specific antibodies. RESULTS: Tissue-specific expression of the transgene was variable, with viral gene expression in the kidney at an intermediate level as compared with other tissues. Overall, the level of transgene expression directly correlated with abundance of nuclear factor-kappa B (NF-kappa B) in the nuclear extracts. High expressing tissue, however, had a constitutively active form of NF kappa B. In contrast, the kidney contained an inducible NF-kappa B, which bound the LTR in combination with Sp1, suggesting a requirement for an activating event in renal HIV-1 expression of the LTR. CONCLUSIONS: These studies indicate that the regulation of the HIV-1 LTR in the kidney is similar to lymphoid tissues, and may explain, in part, why the HIV-1 life cycle is supported in kidney. PMID- 11380820 TI - Hypertonic activation of the renal betaine/GABA transporter is microtubule dependent. AB - BACKGROUND: Epithelial cells in the renal inner medulla accumulate osmolytes such as betaine to maintain normal cell volume during prolonged extracellular hypertonic stress. Betaine accumulation is the result of activation of transcription of the BGT1 transporter gene followed by increased betaine transport. METHODS: We studied the possible role of microtubules in this adaptive mechanism using renal cells in culture. RESULTS.: In cultured renal cell lines [Madin-Darby canine kidney (MDCK) and mouse inner medullary collecting duct (mIMCD-3)], up-regulation of BGT1 activity was maximal after 24 to 30 hours in growth medium made hypertonic (510 mOsm/kg) by the addition of sucrose or NaCl. Up-regulation was reversed within 24 to 36 hours after returning cells to isotonic medium. Both cycloheximide (20 micromol/L) and nocodazole (20 micromol/L) blocked the hypertonic up-regulation of BGT1. Nocodazole was partially effective even when added 16 to 20 hours after the switch to hypertonic medium. Recovery from nocodazole action was rapid, and there was full activation of BGT1 transport within three to six hours after nocodazole removal, suggesting rapid trafficking to the cell surface once microtubules repolymerized. Hypertonic activation of BGT1 transport was detected in an isolated membrane fraction and was blocked by cycloheximide but not by nocodazole. Confocal microscopy confirmed the increased abundance of BGT1 proteins in the plasma membrane of hypertonic cells and showed that BGT1 remained intracellular during nocodazole treatment. CONCLUSIONS: Hypertonic activation of BGT1 in renal cells requires de novo protein synthesis and microtubule-dependent trafficking of additional transporters to the cell surface. The apparent resistance of membrane BGT1 to nocodazole blockade is likely due to the presence in the membrane fraction of an increased intracellular pool of active BGT1 transporters. PMID- 11380821 TI - Down-regulation of hepatic lecithin:cholesterol acyltransferase gene expression in chronic renal failure. AB - BACKGROUND: Chronic renal failure (CRF) is associated with premature arteriosclerosis, impaired high-density lipoprotein (HDL) maturation, increased pre-beta HDL (a lipid-poor HDL species), reduced HDL/total cholesterol ratio, hypertriglyceridemia, and depressed lipolytic activity. The latter has been, in part, attributed to elevated pre-beta HDL, which is a potent inhibitor of lipoprotein lipase (LPL). Accumulation of cholesterol in the arterial wall is a critical step in atherogenesis, and HDL-mediated cholesterol removal from peripheral tissues mitigates atherosclerosis. Lecithin:cholesterol acyltransferase (LCAT) is essential for maturation of HDL and cholesterol removal by HDL from peripheral tissues. Earlier studies have revealed depressed plasma LCAT enzymatic activity in patients with CRF. This study was conducted to determine whether impaired LCAT activity can be confirmed in CRF animals and if so whether it is due to down-regulation of hepatic LCAT expression. METHODS: Hepatic tissue LCAT mRNA and plasma LCAT enzymatic activity were measured in male Sprague-Dawley rats six weeks after excisional 5/6 nephrectomy or sham operation. RESULTS: Compared with the controls, the CRF group exhibited a significant reduction of hepatic tissue LCAT mRNA abundance. The reduction in hepatic LCAT mRNA was accompanied by a marked reduction of plasma LCAT activity and elevation of serum-free cholesterol in the CRF animals. LCAT activity correlated positively with the HDL/total cholesterol ratio and inversely with free cholesterol and triglyceride concentrations. CONCLUSIONS: CRF leads to a marked down-regulation of hepatic LCAT mRNA expression and plasma LCAT activity. This abnormality can impair HDL-mediated cholesterol uptake from the vascular tissue and contribute to cardiovascular disease. In addition, LCAT deficiency can, in part, account for elevated serum-free cholesterol, reduced HDL/total cholesterol, and elevated pre beta HDL in CRF. The latter can, in turn, depress lipolytic activity and hinder triglyceride-rich lipoprotein clearance in CRF. PMID- 11380822 TI - Cellular localization of the potassium channel Kir7.1 in guinea pig and human kidney. AB - BACKGROUND: K(+) channels have important functions in the kidney, such as maintenance of the membrane potential, volume regulation, recirculation, and secretion of potassium ions. The aim of this study was to obtain more information on the localization and possible functional role of the inwardly rectifying K(+) channel, Kir7.1. METHODS: Kir7.1 cDNA (1114 bp) was isolated from guinea pig kidney (gpKir7.1), and its tissue distribution was analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, a genomic DNA fragment (6153 bp) was isolated from a genomic library. cRNA was expressed in Xenopus laevis oocytes for functional studies. Immunohistochemistry and RT-PCR were used to localize Kir7.1 in guinea pig and human kidney. RESULTS: The expression of gpKir7.1 in Xenopus laevis oocytes revealed inwardly rectifying K(+) currents. The reversal potential was strongly dependent on the extracellular K(+) concentration, shifting from -14 mV at 96 mmol/L K(+) to -90 mV at 1 mmol/L K(+). gpKir7.1 showed a low affinity for Ba(2+). Significant expression of gpKir7.1 was found in brain, kidney, and lung, but not in heart, skeletal muscle, liver, or spleen. Immunocytochemical detection in guinea pig identified the gpKir7.1 protein in the basolateral membrane of epithelial cells of the proximal tubule. RT-PCR analysis identified strong gpKir7.1 expression in the proximal tubule and weak expression in glomeruli and thick ascending limb. In isolated human tubule fragments, RT-PCR showed expression in proximal tubule and thick ascending limb. CONCLUSION: Our results suggest that Kir7.1 may contribute to basolateral K(+) recycling in the proximal tubule and in the thick ascending limb. PMID- 11380823 TI - Paracellin-1 is critical for magnesium and calcium reabsorption in the human thick ascending limb of Henle. AB - BACKGROUND: A new protein, named paracellin 1 (PCLN-1), expressed in human thick ascending limb (TAL) tight junctions, possibly plays a critical role in the control of magnesium and calcium reabsorption, since mutations of PCLN-1 are present in the hypomagnesemia hypercalciuria syndrome (HHS). However, no functional experiments have demonstrated that TAL magnesium and calcium reabsorption were actually impaired in patients with HHS. METHODS: Genetic studies were performed in the kindred of two unrelated patients with HHS. Renal magnesium and calcium reabsorption in TAL were analyzed in one homozygous affected patient of each family, one patient with extrarenal hypomagnesemia (ERH), and two control subjects (CSs). RESULTS: We found two yet undescribed mutations of PCLN-1 (Gly 162 Val, Ala 139 Val). In patients with HHS, renal magnesium and calcium reabsorptions were impaired as expected; NaCl renal conservation during NaCl deprivation and NaCl tubular reabsorption in diluting segment were intact. Furosemide infusion in CS markedly increased NaCl, Mg, and Ca urinary excretion rates. In HHS patients, furosemide similarly increased NaCl excretion, but failed to increase Mg and Ca excretion. Acute MgCl(2) infusion in CS and ERH patient provoked a dramatic increase in urinary calcium excretion without change in NaCl excretion. When combined with MgCl(2) infusion, furosemide infusion remained able to induce normal natriuretic response, but was unable to increase urinary magnesium and calcium excretion further. In HHS patients, calciuric response to MgCl(2) infusion was blunted. CONCLUSION: This study is the first to our knowledge to demonstrate that homozygous mutations of PCLN-1 result in a selective defect in paracellular Mg and Ca reabsorption in the TAL, with intact NaCl reabsorption ability at this site. In addition, the study supports a selective physiological effect of basolateral Mg(2+) and Ca(2+) concentration on TAL divalent cation paracellular permeability, that is, PCLN-1 activity. PMID- 11380824 TI - Compensatory up-regulation of angiotensin II subtype 1 receptors in alpha ENaC knockout heterozygous mice. AB - BACKGROUND: In mice, a partial loss of function of the epithelial sodium channel (ENaC), which regulates sodium excretion in the distal nephron, causes pseudohypoaldosteronism, a salt-wasting syndrome. The purpose of the present experiments was to examine how alpha ENaC knockout heterozygous (+/-) mice, which have only one allele of the gene encoding for the alpha subunit of ENaC, control their blood pressure (BP) and sodium balance. METHODS: BP, urinary electrolyte excretion, plasma renin activity, and urinary adosterone were measured in wild type (+/+) and heterozygous (+/-) mice on a low, regular, or high sodium diet. In addition, the BP response to angiotensin II (Ang II) and to Ang II receptor blockade, and the number and affinity of Ang II subtype 1 (AT1) receptors in renal tissue were analyzed in both mouse strains on the three diets. RESULTS: In comparison with wild-type mice (+/+), alpha ENaC heterozygous mutant mice (+/-) showed an intact capacity to maintain BP and sodium balance when studied on different sodium diets. However, no change in plasma renin activity was found in response to changes in sodium intake in alpha ENaC +/- mice. On a normal salt diet, heterozygous mice had an increased vascular responsiveness to exogenous Ang II (P < 0.01). Moreover, on a normal and low sodium intake, these mice exhibited an increase in the number of AT1 receptors in renal tissues; their BP lowered markedly during the Ang II receptor blockade (P < 0.01) and there was a clear tendency for an increase in urinary aldosterone excretion. CONCLUSIONS: alpha ENaC heterozygous mice have developed an unusual mechanism of compensation leading to an activation of the renin-angiotensin system, that is, the up regulation of AT1 receptors. This up-regulation may be due to an increase in aldosterone production. PMID- 11380825 TI - Mycophenolate mofetil prevents salt-sensitive hypertension resulting from angiotensin II exposure. AB - BACKGROUND: Interstitial mononuclear cell infiltration is a feature of experimental models of salt-sensitive hypertension (SSHTN). Since several products of these cells are capable of modifying local vascular reactivity and sodium reabsorption, we investigated whether mycophenolate mofetil (MMF), a drug known to inhibit infiltration and proliferation of immune cells, would modify the SSHTN induced by angiotensin II (Ang II) infusion. METHODS: Sprague-Dawley rats received Ang II for two weeks using subcutaneous minipumps. A high-sodium (4% NaCl) diet was started on the third week and was maintained until the eighth week. MMF (30 mg/kg, N = 15), an immunosuppressive drug, or vehicle (N = 15) was given daily by gastric gavage during the initial three weeks. Sham-operated rats (N = 9) were used as controls. Body weight, blood pressure (tail-cuff plethysmography), and serum creatinine were determined weekly. Urinary malondialdehyde (MDA) excretion, renal histology, and immunohistology, including the presence of Ang II and superoxide-producing cells, were analyzed at the end of Ang II infusion and at eight weeks. RESULTS: MMF treatment did not modify hypertension induced during exogenous Ang II infusion, but prevented the subsequent SSHTN. Tubulointerstitial injury resulting from Ang II infusion was significantly reduced by MMF treatment, as were proliferative activity, T-cell infiltration and activation (interleukin-2 receptor expression), superoxide producing cells, and urinary MDA excretion. Ang II-producing cells were present in the renal tubulointerstitium of rats with SSHTN (60 +/- 30 Ang II-positive cells/mm(2) at 8 weeks) and were reduced by two thirds in the MMF-treated group. Forty percent of lymphocytes infiltrating the tubulointerstitium stained positive for Ang II. The expression of Ang II receptors in the kidney was unmodified. CONCLUSIONS: SSHTN resulting from Ang II infusion is associated with infiltration and activation of immune cells that produce Ang II. MMF treatment reduces these features and prevents the development of SSHTN. PMID- 11380826 TI - Renal phenotype of low kallikrein rats. AB - BACKGROUND: Renal kallikrein has been linked with inheritance of arterial hypertension and with sensitivity to drug nephrotoxicity. Identification of a cause--effect relationship between low kallikrein and intermediate phenotypes has been hampered by the lack of adequate animal models. METHODS: Kallikrein was measured in tissues obtained from rats inbred for low urinary kallikrein excretion (LKR) and wild-type controls. Blood pressure and indices of myocardial contractility were recorded via an intraventricular cannula connected to a transducer. The functional relevance of endogenous angiotensin II (Ang II) in LKR was explored by determining the effect of Ang II subtype 1 (AT(1)) receptor blockade on glomerular filtration rate, renal blood flow, and urinary sodium excretion. In addition, sensitivity to gentamycin-induced nephrotoxicity was evaluated. RESULTS: Kallikrein activity was reduced by 60% in the kidney of LKR (P < 0.01), whereas it was increased in the heart (P < 0.05) and was unaltered in the pancreas, liver, and salivary glands. Heart rate and myocardial contractility were reduced, and the mean blood pressure (MBP) was increased in LKR as compared with controls (P < 0.05). LKR exhibited polydipsia, polyuria, glomerular hyperfiltration, and reduced fractional sodium excretion under basal conditions and impaired renal vasodilation in response to volume expansion. These functional alterations were significantly attenuated by AT(1) receptor blockade. Gentamycin reduced the glomerular filtration rate in LKR, but not in controls. CONCLUSIONS: In LKR, unopposed activity of Ang II appears to be responsible for increased glomerular hydrostatic pressure and augmented tubular reabsorption. Balance between the kallikrein-kinin and renin-angiotensin systems is essential for normal renal function. PMID- 11380827 TI - Endotoxemic renal failure in mice: Role of tumor necrosis factor independent of inducible nitric oxide synthase. AB - BACKGROUND: Renal failure is a frequent complication of sepsis with a high mortality. Tumor necrosis factor (TNF) has been suggested to be a factor in the acute renal failure in sepsis or endotoxemia. Recent studies also suggest involvement of nitric oxide (NO), generated by inducible NO synthase (iNOS), in the pathogenesis of endotoxin-induced renal failure. The present study tested the hypothesis that the role of TNF in endotoxic renal failure is mediated by iNOS derived NO. METHODS: Renal function was evaluated in endotoxemic [Escherichia coli lipopolysaccharide (LPS), 5 mg/kg IP] wild-type and iNOS knockout mice. The effect of TNF neutralization on renal function during endotoxemia in mice was assessed by a TNF-soluble receptor (TNFsRp55). RESULTS: An injection of LPS to wild-type mice resulted in a 70% decrease in glomerular filtration rate (GFR) and in a 40% reduction in renal plasma flow (RPF) 16 hours after the injection. The results occurred independent of hypotension, morphological changes, apoptosis, and leukocyte accumulation. In mice pretreated with TNFsRp55, only a 30% decrease in GFR without a significant change in RPF in response to LPS, as compared with vehicle-treated mice, was observed. Also, the serum NO concentration was significantly lower in endotoxemic wild-type mice pretreated with TNFsRp55, as compared with untreated endotoxemic wild-type mice (260 +/- 52 vs. 673 +/- 112 micromol/L, P < 0.01). In LPS-injected iNOS knockout mice and wild-type mice treated with a selective iNOS inhibitor, 1400W, the development of renal failure was similar to that in wild-type mice. As in wild-type mice, TNFsRp55 significantly attenuated the decrease in GFR (a 33% decline, as compared with 75% without TNFsRp55) without a significant change in RPF in iNOS knockout mice given LPS. CONCLUSIONS: These results demonstrate a role of TNF in the early renal dysfunction (16 h) in a septic mouse model independent of iNOS, hypotension, apoptosis, leukocyte accumulation, and morphological alterations, thus suggesting renal hypoperfusion secondary to an imbalance between, as yet to be defined, renal vasoconstrictors and vasodilators. PMID- 11380828 TI - Total body water reference values and prediction equations for adults. AB - BACKGROUND: The clinical interpretation of total body water (TBW) necessitates the availability of timely comparative reference data. The prediction of TBW volume in renal disease is critical in order to prescribe and monitor the dose of dialysis in the determination of Kt/V. In clinical practice, urea distribution (V) is commonly predicted from anthropometric equations that are several decades old and for white patients only. This article presents new reference values and prediction equations for TBW from anthropometry for white and black adults. METHODS: The study sample included four data sets, two from Ohio and one each from New Mexico and New York, for a total of 604 white men, 128 black men, 772 white women, and 191 black women who were 18 to 90 years of age. The TBW concentration was measured by the deuterium or tritium oxide dilution method, and body composition was measured with a Lunar DXA machine. An all-possible-subsets of regression was used to predict TBW. The accuracy of the selected equations was confirmed by cross-validation. RESULTS: Blacks had larger TBW means than whites at all age groups. The 75th TBW percentile for whites approximated the TBW median for blacks at most ages. The white men and black men and women had the largest TBW means ever reported for healthy individuals. The race- and sex-specific TBW prediction equations included age, weight, and stature, with body mass index (BMI) substituted for weight in the white men. The root mean square errors (RMSEs) and standard errors for the individual (SEIs) ranged from approximately 3.8 to 5.0 L for the men and from 3.3 to 3.6 L for the women. In both men and women, high values of TBW were associated with high levels of total body fat (TBF) and fat-free mass (FFM). CONCLUSION: : TBW in these healthy adults is relatively stable through a large portion of adulthood. There are significant race and sex differences in TBW. These accurate and precise equations for TBW provide a useful tool for the clinical prediction of TBW in renal disease for white and black adults. These are the first TBW prediction equations that are specific for blacks. PMID- 11380829 TI - Prevalence and determinants of erectile dysfunction in hemodialysis patients. AB - BACKGROUND: The prevalence of erectile dysfunction (ED) among patients with end stage renal disease (ESRD) is not known. METHODS: A cross-sectional study was conducted to determine the prevalence of ED among a community-based hemodialysis (HD) population using a two-stage cluster random sampling design. The presence and severity of ED were assessed among 302 ESRD patients using the self administered International Index of Erectile Function-5 (IIEF-5). Logistic regression was used to examine and test associations between ED and other medical conditions. RESULTS: The prevalence of any level of ED was 82% (95% CI, 76 to 87%) for all HD subjects. The prevalence of severe ED was 45% (CI, 36 to 55%). Subjects younger than 50 years had a prevalence of ED of 63% (CI, 53 to 71%), while in subjects 50 years or older, it was 90% (CI, 84 to 94%). A multivariable analysis demonstrated increasing age (50 to 59, OR = 2.04, 95% CI, 1.3 to 3.1; 60 to 69, OR = 5.5, 95% CI, 1.9 to 15.6) and diabetes (OR = 2.0, 95% CI, 1.2 to 3.3) to be independently associated with the presence of any level of ED. However, neither the subjects' age nor history of diabetes predicted the severity of ED among subjects with ED. The use of angiotensin-converting enzyme inhibitors (ACEIs) was inversely associated with ED (OR = 0.41, 95% CI, 0.17 to 0.98). Poor functional status (Karnofsky score or the Index of Physical Impairment) was not associated with ED. CONCLUSIONS: ED is extremely prevalent among HD patients. Increasing age, diabetes, and nonuse of ACEIs were associated with higher prevalence of ED. The high prevalence of ED was seen even among patients with good functional status. PMID- 11380830 TI - Dimethylglycine accumulates in uremia and predicts elevated plasma homocysteine concentrations. AB - BACKGROUND: Hyperhomocysteinemia is a risk factor for atherosclerosis that is common in chronic renal failure (CRF), but its cause is unknown. Homocysteine metabolism is linked to betaine-homocysteine methyl transferase (BHMT), a zinc metalloenzyme that converts glycine betaine (GB) to N,N dimethylglycine (DMG). DMG is a known feedback inhibitor of BHMT. We postulated that DMG might accumulate in CRF and contribute to hyperhomocysteinemia by inhibiting BHMT activity. METHODS: Plasma and urine concentrations of GB and DMG were measured in 33 dialysis patients (15 continuous ambulatory peritoneal dialysis and 18 hemodialysis), 33 patients with CRF, and 33 age-matched controls. Concentrations of fasting plasma total homocysteine (tHcy), red cell and serum folate, vitamins B(6) and B(12), serum zinc, and routine biochemistry were also measured. Groups were compared, and determinants of plasma tHcy were identified by correlations and stepwise linear regression. RESULTS: Plasma DMG increased as renal function declined and was twofold to threefold elevated in dialysis patients. Plasma GB did not differ between groups. The fractional excretion of GB (FE(GB)) was increased tenfold, and FED(MG) was doubled in CRF patients compared with controls. Plasma tHcy correlated positively with plasma DMG, the plasma DMG:GB ratio, plasma creatinine, and FE(GB) and negatively with serum folate, zinc, and plasma GB. In the multiple regression model, only plasma creatinine, plasma DMG, or the DMG:GB ratio was independent predictors of tHcy. CONCLUSIONS: DMG accumulates in CRF and independently predicts plasma tHcy concentrations. These findings suggest that reduced BHMT activity is important in the pathogenesis of hyperhomocysteinemia in CRF. PMID- 11380831 TI - Sensitivity to meat protein intake and hyperoxaluria in idiopathic calcium stone formers. AB - BACKGROUND: High protein intake is an accepted risk factor for renal stone disease. Whether meat protein intake affects oxaluria, however, remains controversial in healthy subjects and in stone formers. This study was designed (1) to test the oxaluric response to a meat protein load in male recurrent idiopathic calcium stone formers (ICSFs) with and without mild metabolic hyperoxaluria (MMH and non-MMH, respectively), as well as in healthy controls, and (2) to seek for possible disturbed vitamin B(6) metabolism in MMH, in analogy with primary hyperoxaluria. METHODS: Twelve MMH, 8 non-MMH, and 13 healthy males were studied after five days on a high meat protein diet (HPD; 700 g meat/fish daily) following a run-in phase of five days on a moderate protein diet (MPD; 160 g meat/fish daily). In both diets, oxalate-rich nutrients were avoided, as well as sweeteners and vitamin C-containing medicines. Twenty-four-hour urinary excretion of oxalate was measured on the last day of each period, along with 4 pyridoxic acid (U(4PA)) and markers of protein intake, that is, urea, phosphate, uric acid, and sulfate. Serum pyridoxal 5' phosphate (S(P5P)) was measured after protein loading. RESULTS: Switching from MPD (0.97 +/- 0.18 g protein/kg/day) to HPD (2.26 +/- 0.38 g protein/kg/day) led to the expected rise in the urinary excretion rates of all markers of protein intake in all subjects. Concurrently, the mean urinary excretion of oxalate increased in ICSFs taken as a whole (+73 +/ 134 micromol/24 h, P = 0.024) as well as in the MMH subgroup (+100 +/- 144 micromol/24 h, P = 0.034) but not in controls (-17 +/- 63 micromol/24 h). In seven ICSFs (4 MMH and 3 non-MMH) but in none of the healthy controls (P = 0.016, chi square), an increment in oxaluria was observed and considered as significant based on the intra-assay coefficient of variation at our laboratory (8.5%). There was no difference in S(P5P)nd U(4PA)etween the groups after protein loading. CONCLUSION: Approximately one third of ICSFs with or without so-called MMH are sensitive to meat protein in terms of oxalate excretion, as opposed to healthy subjects. Mechanisms underlying this sensitivity to meat protein remain to be elucidated and do not seem to involve vitamin B(6) deficiency. PMID- 11380832 TI - Add-on angiotensin receptor blockade with maximized ACE inhibition. AB - BACKGROUND: Prolonged angiotensin-converting enzyme (ACE) inhibitor therapy leads to angiotensin I (Ang I) accumulation, which may "escape" ACE inhibition, generate Ang II, stimulate the Ang II subtype 1 (AT1) receptor, and exert deleterious renal effects in patients with chronic renal diseases. We tested the hypothesis that losartan therapy added to a background of chronic (>3 months) maximal ACE inhibitor therapy (lisinopril 40 mg q.d.) will result in additional Ang II antagonism in patients with proteinuric chronic renal failure with hypertension. METHODS: Sixteen patients with proteinuric moderately advanced chronic renal failure completed a two-period, crossover, randomized controlled trial. Each period was one month with a two-week washout between periods. In one period, patients received lisinopril 40 mg q.d. along with other antihypertensive therapy, and in the other, losartan 50 mg q.d. was added to the previously mentioned regimen. Hemodynamic measurements included ambulatory blood pressure monitoring (ABP; Spacelabs 90207), glomerular filtration rate (GFR) with iothalamate clearances and cardiac outputs by acetylene helium rebreathing technique. Supine plasma renin activity and plasma aldosterone and 24-hour urine protein were measured in all patients. RESULTS: Twelve patients had diabetic nephropathy, and four had chronic glomerulonephritis. The mean age (+/- SD) was 53 +/- 9 years. The body mass index was 38 +/- 5.7 kg/m(2), and all except two patients were males. Seated cuff blood pressure was 156 +/- 18/88 +/- 12 mm Hg. The pulse rate was 77 +/- 11 per min, and the cardiac index was 2.9 +/- 0.5 L/min/m(2). Mean log 24-hour protein excretion/g creatinine or overall ABPs did not change. Mean placebo subtracted, losartan-attributable change in protein excretion was +1% (95% CI, -20% to 28%, P = 0.89). Similarly, the change in systolic ambulatory blood pressure (ABP) was 4.6 mm Hg (-5.7 to 14.9, P = 0.95), and diastolic ABP was 1.5 mm Hg (-4.5 to 7.6, P = 0.59). No change was seen in cardiac output. However, there was a mean 14% increase (95% CI, 3 to 26%, P = 0.017) in GFR attributable to losartan therapy. A concomitant fall in plasma renin activity by 32% was seen (95% CI, -15%, - 45%, P = 0.002). No hyperkalemia, hypotension, or acute renal failure occurred in the trial. These results were not attributable to sequence or carryover effects. CONCLUSIONS: Add-on losartan therapy did not improve proteinuria or ABP over one month of add on therapy. Improvement of GFR and fall in plasma renin activity suggest that renal hemodynamic and endocrine changes are more sensitive measures of AT1 receptor blockade. Whether add-on AT1 receptor blockade causes antiproteinuric effects or long-term renal protection requires larger and longer prospective, randomized controlled trials. PMID- 11380833 TI - Twenty-four-hour urine chemistries and the risk of kidney stones among women and men. AB - BACKGROUND: Results of a 24-hour urine collection are integral to the selection of the most appropriate intervention to prevent kidney stone recurrence. However, the currently accepted definitions of normal urine values are not firmly supported by the literature. In addition, little information is available about the relationship between risk of stone formation and the levels of urinary factors. Unfortunately, the majority of previous studies of 24-hour urine chemistries were limited by the inclusion of recurrent stone formers and poorly defined controls. METHODS: We obtained 24-hour urine collections from 807 men and women with a history of kidney stone disease and 239 without a history who were participants in three large ongoing cohort studies: the Nurses' Health Study I (NHS I; mean age of 61 years), the Nurses' Health Study II (NHS II; mean age of 42 years), and the Health Professionals Follow-up Study (HPFS; mean age of 59 years). RESULTS: Mean 24-hour urine calcium excretion was higher and urine volume was lower in cases than controls in NHS I (P < or = 0.01), NHS II (P < or = 0.13) and HPFS (P < or = 0.01), but urine oxalate and citrate did not differ. Among women, urine uric acid was similar in cases and controls but was lower in cases in men (P = 0.06). The frequency of hypercalciuria was higher among the cases in NHS I (P = 0.26), NHS II (P = 0.03), and HPFS (P = 0.02), but 27, 17, and 14% of the controls, respectively, also met the definition of hypercalciuria. The frequency of hyperoxaluria did not differ between cases and controls, but was three times more common among men compared with women. After adjusting for the other urinary factors, the relative risk of stone formation increased with increasing urine calcium levels and concentration in all three cohorts but not in a linear fashion. Compared with individuals with a urine calcium concentration of <75 mg/L, the relative risk of stone formation among those with a urine calcium concentration of > or =200 mg/L for NHS I was 4.34 (95% CI, 1.59 to 11.88), for NHS II was 51.09 (4.27 to 611.1), and for HPFS was 4.30 (1.71 to 10.84). There was substantial variation in the relative risks for stone formation for the concentration of other urine factors within the different cohorts. CONCLUSIONS: The traditional definitions of normal 24-hour urine values need to be reassessed, as a substantial proportion of controls would be defined as abnormal, and the association with risk of stone formation may be continuous rather than dichotomous. The 24-hour urine chemistries are important for predicting risk of stone formation, but the significance and the magnitudes of the associations appear to differ by age and gender. PMID- 11380834 TI - Plasma proteins containing damaged L-isoaspartyl residues are increased in uremia: implications for mechanism. AB - BACKGROUND: Several alterations of protein structure and function have been reported in uremia. Impairment of a transmethylation-dependent protein repair mechanism possibly related to a derangement in homocysteine metabolism is also present in this condition, causing erythrocyte membrane protein damage. Homocysteine may affect proteins via the accumulation of its parent compound S adenosylhomocysteine (AdoHcy), a powerful in vivo methyltransferase inhibitor. However, since plasma homocysteine is mostly protein bound, a direct influence on protein structures cannot be ruled out. We measured the levels of L-isoaspartyl residues in plasma proteins of uremic patients on hemodialysis. These damaged residues are markers of molecular age, which accumulate when transmethylation dependent protein repair is inhibited and/or protein instability is increased. METHODS: L-isoaspartyl residues in plasma proteins were quantitated using human recombinant protein carboxyl methyl transferase (PCMT). Plasma concentrations of homocysteine metabolites were also measured under different experimental conditions in hemodialysis patients. RESULTS: The concentration of damaged plasma proteins was increased almost twofold compared to control (controls 147.83 +/- 17.75, uremics 282.80 +/- 26.40 pmol of incorporated methyl groups/mg protein, P < 0.003). The major protein involved comigrated with serum albumin. Although hyperhomocysteinemia caused a redistribution of thiols bound to plasma proteins, this mechanism did not significantly contribute to the increase in isoaspartyl residues. The S-adenosylmethionine (AdoMet)/AdoHcy concentration ratio, an indicator of the flux of methyl group transfer, was altered. This ratio was partially corrected by folate treatment (0.385 +/- 0.046 vs. 0.682 +/- 0.115, P < 0.01), but protein L-isoaspartate content was not. CONCLUSIONS: Plasma protein damage, as determined by protein L-isoaspartyl content, is increased in uremia. This alteration is to be ascribed to an increased protein structural instability, rather than the effect of hyperhomocysteinemia. PMID- 11380835 TI - Clinical course of peritonitis due to Pseudomonas species complicating peritoneal dialysis: a review of 104 cases. AB - BACKGROUND: Peritonitis due to Pseudomonas species is a serious complication in continuous ambulatory peritoneal dialysis (CAPD) patients. The clinical course of peritonitis due to Pseudomonas complicating CAPD remains unclear. METHODS: All of the Pseudomonas species episodes of peritonitis in our dialysis unit were studied from 1995 to 1999. During this period, there were 859 episodes of peritonitis recorded, 113 of which were caused by the Pseudomonas species. Nine episodes were excluded because they were mixed growth. The remaining 104 episodes in 68 patients were reviewed. RESULTS: The underlying renal diagnosis and prevalence of comorbid conditions of the 68 patients were similar to those found in our entire dialysis population. There was a history of antibiotic therapy within 30 days of the onset of peritonitis due to the Pseudomonas species in 69 episodes (66.3%). In 47 episodes (45.2%) there was a concomitant exit site infection. The overall primary response rate was 60.6% and the complete cure rate was 22.1%. The presence of exit site infection was associated with a lower primary response rate (22 in 47 vs. 41 in 57 episodes, P < 0.01) and a lower complete cure rate (5 in 47 vs. 18 in 57 episodes, P < 0.02). The episodes that had received recent antibiotic therapy had a significantly lower complete cure rate than the de novo cases (8 in 69 vs. 15 in 35 episodes, P < 0.001). Episodes receiving third generation cephalosporin as part of the initial antibiotic regimen had a significantly higher primary response rate than the ones that initially received aminoglycoside (54 in 81 episodes vs. 8 in 22 episodes, P < 0.05), but their complete cure rates were similar. Twenty-four cases failed to respond to antibiotics and the Tenckhoff catheter was removed. The chance of returning to CAPD was higher when the Tenckhoff catheter was removed on day 10 than on day 15 (9 in 14 cases vs. 5 in 10 cases), although the result was not statistically significant. The Tenckhoff catheter was removed and replaced at another site simultaneously in another 14 cases after the effluent cleared up. None of these patients had a relapse of peritonitis within three months. CONCLUSIONS: Recent antibiotic therapy is the major risk factor for peritonitis due to the Pseudomonas species. Exit site infection and recent antibiotic therapy are associated with poor therapeutic response to antibiotics. When the therapeutic response is suboptimal, early Tenckhoff catheter removal may help preserve the peritoneum for further peritoneal dialysis. Elective Tenckhoff catheter exchange after clearing up the peritoneal dialysis effluent may also reduce the likelihood of relapse. It is desirable to use third-generation cephalosporin in the initial antibiotic regimen for peritonitis treatment in localities with a high incidence of peritonitis due to the Pseudomonas species. PMID- 11380836 TI - Dipyridamole inhibits human peritoneal mesothelial cell proliferation in vitro and attenuates rat peritoneal fibrosis in vivo. AB - BACKGROUND: Peritoneal fibrosis (PF) is one of the most serious complications after long-term continuous ambulatory peritoneal dialysis (CAPD). Proliferation of human peritoneal mesothelial cells (HPMC) and matrix over-production are regarded as the main processes predisposing to PF. Dipyridamole (DP) has been reported to have potential as an antiproliferative and antifibrotic agent. We thus investigated the effect of DP in inhibiting proliferation and collagen synthesis of HPMC. A rat model of peritonitis-induced PF was also established to demonstrate the in vivo preventive effect of DP. METHODS: HPMC was cultured from human omentum by an enzyme digestion METHOD: Cell proliferation was measured by the methyltetrazolium assay. Intracellular cAMP was measured using an enzyme immunoassay (EIA) kit. Total collagen synthesis was measured by (3)H-proline incorporation assay. Expression of collagen alpha1 (I) and collagen alpha 1 (III) mRNAs was determined by Northern blotting. The rat model of peritonitis-induced PF was developed by adding dextran microbeads (Cytodex, 8 mg/1 mL volume) to a standardized suspension (3 x 10(9)) of Staphylococcus aureus. DP was administrated via intravenous infusion (4 mg in 1 h) daily for seven days. Macroscopic grading of intraperitoneal adhesions and histological analyses of peritoneal thickness and collagen expression were performed. RESULTS: Addition of DP to HPMC cultures suppressed serum-stimulated cell proliferation and collagen synthesis. The antimitogenic and antifibrotic effects of DP appear to be predominantly mediated through the cAMP pathway, as DP increased intracellular cAMP in a dose-dependent manner. The macroscopic grade of intraperitoneal adhesion and peritoneal thickness were both significantly increased in animals treated with Cytodex plus S. aureus; on the other hand, DP attenuated these fibrotic changes with statistical significance (P < 0.01). Analysis of gene expression of collagen alpha 1 (I) and alpha1 (III) in the peritoneal tissue of experimental animals yielded similar results. CONCLUSIONS: This study suggests that dipyridamole may have therapeutic potential in treating peritoneal fibrosis. PMID- 11380837 TI - Venous neointimal hyperplasia in polytetrafluoroethylene dialysis grafts. AB - BACKGROUND: Vascular access dysfunction is the most important cause of morbidity and hospitalization in the hemodialysis population in the United States at a cost of $1 billion per annum. Venous neointimal hyperplasia (VNH) characterized by stenosis and subsequent thrombosis accounts for the overwhelming majority of pathology resulting in polytetrafluoroethylene (PTFE) dialysis graft failure. Despite the magnitude of the problem and the enormity of the cost ($1 billion), there are currently no effective therapies for the prevention or treatment of venous neointimal hyperplasia in PTFE dialysis grafts. METHODS: Tissue samples were collected from the graft-vein anastomosis of stenotic PTFE grafts during surgical revision. Specimens were graded using standard light microscopy and immunohistochemistry for the magnitude of neointimal hyperplasia and for the expression of specific cell types, cytokines, and matrix proteins. RESULTS: VNH was characterized by the (1) presence of smooth muscle cells/myofibroblasts, (2) accumulation of extracellular matrix components, (3) angiogenesis within the neointima and adventitia, and (4) presence of an active macrophage cell layer lining the PTFE graft material. Platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were expressed by smooth muscle cells/myofibroblasts within the venous neointima, by macrophages lining both sides of the PTFE graft, and by vessels within the neointima and adventitia. CONCLUSIONS: Our results suggest that macrophages, specific cytokines (bFGF, PDGF, and VEGF), and angiogenesis within the neointima and adventitia are likely to contribute to the pathogenesis of VNH in PTFE dialysis grafts. Interventions aimed at these specific mediators and processes may be successful in reducing the very significant human and economic costs of vascular access dysfunction. PMID- 11380838 TI - Assessment of a policy to reduce placement of prosthetic hemodialysis access. AB - BACKGROUND: The aim of this study was to evaluate the determinants of access patency and revision, including the effects of reducing the placement of prosthetic hemodialysis access. METHODS: A retrospective cohort study of all hemodialysis accesses placed at the Veteran's Administration Puget Sound Health Care System between 1992 and 1999 was conducted. A policy was instituted in 1996 that maximized the use of autogenous hemodialysis access. The impacts of the policy change, demographics, and comorbid factors on access type and patency, were examined. Primary and secondary patency rates were examined using the Kaplan -Meier method, and factors associated with failure and revision were examined using Cox proportional hazard models and Poisson regression. RESULTS: During the study, 104 accesses (61 prosthetic grafts and 43 autogenous fistulas) were placed prior to 1996, and 118 (31 prosthetic grafts and 87 autogenous fistulas) were placed after 1996. There was a significant increase in autogenous fistulas placed after 1996 (87 out of 118) compared with before 1996 (43 out of 104, P < 0.001). At one year, autogenous fistulas demonstrated superior primary patency (56 vs. 36%, P = 0.001) and secondary patency (72 vs. 58%, P = 0.003) compared with prosthetic grafts. After adjustment for age, race, side of access placement, and history of prior access placement, patients with a prosthetic graft were estimated to experience a 78% increase in the risk of primary access failure when compared with similar patients having an autogenous access [adjusted relative risk (aRR) = 1.78, 95% CI 1.21--2.62, P = 0.003)]. Similarly, the adjusted relative risk of secondary access failure for comparing prosthetic grafts with autogenous fistulas was estimated to be 2.21 (95% CI 1.38--3.54, P = 0.001). The adjusted risk of access revision was 2.89-fold higher for prosthetic grafts than for autogenous fistulas (95% CI 1.88--4.44, P < 0.001). CONCLUSIONS: Autogenous conduits demonstrated superior performance when compared with prosthetic grafts in terms of primary and secondary patency and number of revisions. A policy emphasizing the preferential placement of autogenous fistulas over prosthetic grafts may result in improved patency and a reduction in the number of procedures required to maintain dialysis access patency. PMID- 11380840 TI - Ultrasound dilution evaluation of pediatric hemodialysis vascular access. AB - BACKGROUND: Hemodialysis access thrombosis is a significant cause of morbidity for hemodialysis patients and results from decreased access flow caused by venous outflow tract stenosis. Ultrasound dilution (UD) is a practical, noninvasive, and reliable indicator of access flow and is effective in predicting venous stenosis in adult patients receiving hemodialysis. METHODS: The current study is the first to our knowledge to evaluate the accuracy of UD in predicting hemodialysis access stenosis in a pediatric hemodialysis population. Thirteen pediatric patients receiving hemodialysis via permanent access (4 AVF and 9 AVG) received 73 UD measurements over three months. RESULTS: Mean raw access flow (QA) was 720 +/- 428 mL/min, and mean corrected access flow (QAcorr) was 886 +/- 537 mL/min/1.73 m(2). QAcorr was significantly lower in accesses with stenosis (401 +/- 176 mL/min/1.73 m(2)) versus accesses without stenosis (1158 +/- 330 mL/min/1.73 m(2), P < 0.0001). Unlike flow values reported by raw QA, there was no overlap in flow values reported by QAcorr in accesses with stenosis (174 to 579 mL/min/1.73 m(2)) versus accesses without stenosis (709 to 1711 mL/min/1.73 m(2)). Two patients with an AVG who had QAcorr less than 600 mL/min/1.73 m(2) developed an access thrombosis within one week after UD measurement. No patients with QAcorr greater than 700 mL/min/1.73 m(2) developed access thrombosis in the 30 days following UD measurement. CONCLUSIONS: : The current study supports the use of monthly UD measurement to prevent access thrombosis in children receiving hemodialysis. PMID- 11380839 TI - Oxidative modification of low-density lipoproteins and the outcome of renal allografts at 1 1/2 years. AB - BACKGROUND: Previous studies reported a significant association between hyperlipidemia of the recipient and chronic allograft nephropathy (CAN). However, the nature and the pathogenic mechanism of circulating lipid abnormalities in CAN remain unclear. METHODS: In a prospective study of 50 consecutive adult recipients of a cadaveric renal allograft, we investigated the impact of lipid abnormalities on the outcome of the graft at 1 1/2 years. Besides morphometric analysis of implantation and protocol biopsies, clinical and biochemical variables were studied at three-month intervals. Plasma concentrations of oxidized low-density lipoprotein (OxLDL) were determined by means of enzyme linked immunosorbent assay. Immunohistochemical staining for OxLDL and macrophages was performed on paired renal biopsies. Study end points were the fractional interstitial volume and the 24-hour creatinine clearance at 11/2 years. RESULTS: High-density lipoprotein (HDL) cholesterol of the recipient < or =47 mg/dL was a risk factor for the functional (RR = 1.56; 95% CI, 0.978 to 2.497) and the morphological (RR = 2.75; 95% CI, 1.075 to 7.037) outcome of the graft, mainly in patients without acute rejection (RR = 2.03; 95% CI, 1.13 to 3.65, and RR = 4.67; 95% CI, 1.172 to 18.582, respectively). Interstitial accumulation of OxLDL was inversely associated with HDL cholesterol (R = -0.476, P = 0.019), and was associated with a higher density of tubulointerstitial macrophages (R = 0.656, P = 0.001) and a higher fractional interstitial volume at 11/2 years (P = 0.049). CONCLUSION: Decreased HDL cholesterol levels of the recipient adversely affect the outcome of renal allografts through the accumulation of OxLDL in the renal interstitium of the graft. Interstitial accumulation of OxLDL was associated with the presence of macrophages and the development of interstitial fibrosis. PMID- 11380841 TI - Children with hypoalbuminemia on continuous peritoneal dialysis are at risk for technique failure. AB - BACKGROUND: Few data are available on the clinical significance of hypoalbuminemia [serum albumin (SA) <35 g/L] in children with end-stage renal disease (ESRD) on continuous peritoneal dialysis (CPD). This study was conducted to analyze the prevalence of hypoalbuminemia, its predictive factors, and its clinical impact in these children. METHODS: A retrospective analysis was done of 180 patients on CPD over the last 22 years. Patients were excluded from the study if they were on CPD for less than four months or had nephrotic syndrome. Demographic, clinical, and biochemical variables were studied. Children continued on CPD until they received a transplant or were transferred to an adult unit or to hemodialysis as a result of technique failure. The subjects were divided into two groups based on SA levels at last follow-up. RESULTS: A total of 135 children was included. After a mean duration of CPD of 573 +/- 437 (120 to 2960) days, 54 children (40%) were observed to have hypoalbuminemia. Four patients (2.9%) died, 7 (5.2%) continued on continuous cyclic peritoneal dialysis, and 13 (9.6%) were transferred to an adult unit for continuation of CPD. Ninety-five (70.3%) were transplanted, and 16 (11.8%) were transferred to hemodialysis because of technique failure. Children in group I (N = 54, SA <35 g/L), compared with group II (N = 81, SA > or =35 g/L), were younger at initiation of PD, more likely to have hypoalbuminemia at one month and six months after initiation of PD, and have more episodes of peritonitis. No differences were seen between the groups in gender, modality of CPD, body surface area, initial body mass index, and presence of hypertension or acidosis. The only factors predictive of hypoalbuminemia on follow-up were low SA at one month after PD and recurrent peritonitis using multiple logistic regression analysis. Evaluating the clinical impact of hypoalbuminemia, we observed a higher incidence of failed PD in children who had hypoalbuminemia. CONCLUSION: Low SA at one month after PD and recurrent peritonitis are predictive of hypoalbuminemia in children on CPD, which is associated with an increased incidence of CPD failure. PMID- 11380842 TI - Fat-kidneyed. PMID- 11380843 TI - Relationship between nonphenacetin-combined analgesics and nephropathy. PMID- 11380844 TI - Prevalence of autosomal-dominant polycystic kidney disease in Alentejo, Portugal. PMID- 11380845 TI - Post-transplant recurrence of focal segmental glomerulosclerosis. PMID- 11380846 TI - Prealbumin and mortality. PMID- 11380847 TI - 7-84 PTH or aluminum antagonism in the resistance of uremic bone to the remodeling effect of endogenous PTH? PMID- 11380848 TI - Post-transplant hypophosphatemia. PMID- 11380849 TI - PRISMS: new educational strategies for medical education. PMID- 11380850 TI - Square pegs in round holes. PMID- 11380851 TI - Research training in population health. PMID- 11380852 TI - Assessment in postgraduate dental education. PMID- 11380853 TI - Educational implications of practice isolation. PMID- 11380854 TI - Knowledge and practices regarding tuberculosis: a survey of final-year medical students from Canada, India and Uganda. AB - CONTEXT: Tuberculosis is one of the most common infectious diseases worldwide and is responsible for the largest number of deaths from a single infectious cause. OBJECTIVE: The objective of this study was to compare the knowledge of and practices regarding tuberculosis in final-year medical students at schools from endemic and non-endemic areas. SUBJECTS: Final-year medical students at McMaster University in Canada, the Christian Medical College in India, and Makerere University in Uganda. METHODS: A questionnaire consisting of 20 multiple-choice questions assessing knowledge, practices, and exposure. A total knowledge score (maximum=13) and a total practice score (maximum=5) were created for each study site. RESULTS: 160 questionnaires were returned; the response rate was 68.4% (65/95) for McMaster University, 39.7% (23/58) for the Christian Medical College and 78.3% (72/92) for Makerere University. Students from Makerere University had the highest knowledge scores but differences were non-significant after adjustment for patient exposure and curriculum time (F(2,153)= 1.80, P=0.16). Differences in practice scores, however, remained significant after adjusting for curriculum time and patient exposure (F(2,153)=5.14, P=0.006). Knowledge score (F(1,156)=5.05, P=0.02), patient exposure (F(1,153)=9.11, P=0.003), and curriculum time and patient exposure (F(2,153)=5.14, P=0.006) were statistically significant positive predictors of the total practice score. CONCLUSIONS: This study demonstrated significant differences in undergraduate exposure to tuberculosis, total knowledge, and practice competency at three medical schools in Canada, India, and Uganda. In general, the knowledge base and practice competency of all three graduating classes was adequate. PMID- 11380855 TI - Assessment in postgraduate dental education: an evaluation of strengths and weaknesses. AB - This paper describes a study designed to evaluate assessment in postgraduate dental education in England, identifying strengths and weaknesses and focusing specifically on its relevance, consistency and cost-effectiveness. METHODS: A four-phase qualitative method was used: a mapping of current career paths, assessment policy, and issues (phase 1); more detailed studies of the practice of assessment for a range of courses, and the systemic/management perspective of assessment (i.e. quality assurance) (phases 2 and 3), and analysis and reporting (phase 4). Data were analysed from documents, interviews, group consultations and observations. RESULTS AND DISCUSSION: Five key issues may be distilled from the findings: (i) lack of formal assessment of general professional training; (ii) trainer variation in assessment; (iii) the extent to which assessments are appropriate indicators of later success; (iv) the relationship between assessment and patient care, and (v) data to assess the costs of assessment. CONCLUSION: Current assessment procedures might be improved if consideration is given to: assessment which supports an integrated period of general professional training; training for trainers and inspection procedures to address variation; more authentic assessments, based directly on clinical work and grading cases and posts, and better data on allocation of resources, in particular clinicians' time given to assessment. PMID- 11380856 TI - Development and validation of a questionnaire to evaluate the effectiveness of evidence-based practice teaching. AB - AIM: The aim of this study was to develop and validate a questionnaire to evaluate the effectiveness of evidence-based practice (EBP) teaching. METHODS: The 152 questionnaires completed by health care professionals with a range of EBP experience were used in this study. Cronbach's alpha for the knowledge and attitude questions indicated a satisfactory level of internal consistency (i.e. >0.60). RESULTS: The discriminative validity was evidenced by a statistically significant difference in the knowledge and attitude scores of 'novices' (i.e. little or no prior EBP education) compared with 'experts' (i.e. health care professionals and academics currently teaching EBP). Moderate to good (> or =0.4) sensitivity index scores were observed for both knowledge and attitude scores as the result of comparing individuals before and after an EBP intervention. CONCLUSIONS: The results of this validation study indicate that the developed questionnaire is a satisfactory tool with which to evaluate the effectiveness of EBP teaching interventions. PMID- 11380857 TI - Faculty attitudes towards medical communication and their perceptions of students' communication skills training at Dalhousie University. AB - SETTING: Faculty of Medicine, Dalhousie University, Halifax, Nova Scotia, Canada. OBJECTIVES: (1) To assess the attitudes of full-time clinical faculty members towards medical communication using the newly developed Attitudes Towards Medical Communication Scale; (2) to determine faculty members' perceptions of communications training for students and residents. METHODS: An anonymous self completion survey was sent to 233 full-time clinical faculty members. The questionnaire asked about faculty attitudes towards medical communication, and assessed faculty members' views of student and resident training in communication. RESULTS: Faculty scored highly in the Attitudes Towards Medical Communication Scale, with a mean score of 51.5 (SD 4.1) out of a possible 60. In univariate analysis, rating of personal enjoyment of teaching, rating of the importance of teaching, and having attended at least one faculty communications workshop in the previous 5 years were significantly associated with higher scale scores. When these factors were assessed using linear regression, only having attended a workshop and higher rating of the importance of teaching remained significant. Faculty assessed student training in communications skills poorly overall. When assessing seven specific communications areas, more than 20% rated this training as poor for six of the areas for third- and fourth-year students and for five of the areas for residents. CONCLUSIONS: Clinical faculty at Dalhousie have very positive attitudes towards medical communication, and more highly positive attitudes are found in those who have attended a communications workshop. Despite this evidence that faculty appreciate the importance of medical communication skills, many assessed students' training in this curriculum area as poor. PMID- 11380858 TI - A framework for developing excellence as a clinical educator. AB - The current emphasis on providing quality undergraduate and postgraduate medical education has focused attention on the educational responsibilities of all doctors. There is a greater awareness of the need to train doctors as educators and courses have been set up to satisfy this need. Some courses, such as those on how to conduct appraisal, are specific to one task facing a medical educator. Other courses take a broader view and relate educational theory to practice. In this paper we describe an outcome-based approach in which competence in teaching is defined in terms of 12 learning outcomes. The framework provides a holistic approach to the roles of the teacher and supports the professionalism of teaching. Such a framework provides the basis for the development of a curriculum for teaching excellence. It helps to define important competences for different categories of teachers, communicate the areas to be addressed in a course, identify gaps in course provision, evaluate courses, assist in staff planning and allow individuals to assess their personal learning needs. The framework is presented to encourage wider debate. PMID- 11380859 TI - The clinician's role in meeting patient information needs: suggested learning outcomes. AB - BACKGROUND: Patients have always fulfilled their information needs from a variety of different sources over time. Clinician-patient consultations are one part of that process. Some patients have increasing opportunities to obtain information through new sources such as the internet, touch-screens, and patient-held records. Others remain poorly informed. OBJECTIVES: To identify learning outcomes for clinicians in meeting patient information needs and working with well informed patients. DESIGN: Four-stage multicentre multidisciplinary qualitative study. SETTING AND SUBJECTS: (1) Semistructured interviews with 20 clinicians in Glasgow; (2) semistructured interviews with 52 clinicians in Nottingham and London; (3) testing of consensus by postal questionnaire and Delphi method amongst 37 clinicians in medicine, nursing and the professions allied to medicine, and (4) conference to discuss results. RESULTS: 46 learning outcomes were identified in the eight areas of: placing a higher priority on patient information and education; understanding the patient's information needs and environment; understanding the emotional aspects of learning; developing patient understanding; helping patients to understand about health care and health care information; learning from the patient; knowing about information sources and their use, and issues of multidisciplinary working. CONCLUSIONS: The suggested learning outcomes provide the basis for wider discussion, for possible inclusion in curricula both at undergraduate and continuing education levels, and as the basis for the development of new educational materials. PMID- 11380860 TI - Pre-registration house officer rotations incorporating general practice: does the order of rotation matter? AB - CONTEXT: In relation to pre-registration house officer (PRHO) rotations incorporating general practice, previous research has recommended that where possible, no PRHO should undertake general practice as the first placement, because of the difficulties encountered. It was recognized that logistically, this could make such schemes almost unworkable. Within the context of a larger qualitative evaluation comparing how 24 PRHOs learned in hospital and general practice settings, the issue of rotation order was explored. METHODS: In-depth semistructured interviews were conducted with the 12 PRHOs who were involved in general practice rotations. They were interviewed at the beginning and end of the PRHO year, and following their return to hospital work after the general practice placement. RESULTS: Each rotation order had both advantages and disadvantages, with no particular rotation order being obviously better or worse for the PRHOs involved. CONCLUSIONS: This small qualitative evaluation has highlighted a number of advantages and disadvantages specific to each rotation order, and makes some practical recommendations to help alleviate the problems encountered. It is important that future evaluations of similar schemes consider this issue, as there are conflicting reports about the significance of the rotation order. PMID- 11380861 TI - How well prepared are graduates for the role of pre-registration house officer? A comparison of the perceptions of new graduates and educational supervisors. AB - OBJECTIVE: The principal aim of undergraduate medical education is to produce competent pre-registration house officers (PRHOs). We examined and compared the perceptions of graduates and educational supervisors concerning how well prepared graduates were for their first post. METHODS: A postal questionnaire was sent to house officers who had graduated from Manchester 3 months earlier and also to educational supervisors of PRHOs in the North-west Region. The questionnaires were based on the competencies set out by the General Medical Council of the United Kingdom. RESULTS: The response rates were 66% from the graduates and 76% from the supervisors. Of the 18 broad areas of competence listed, only four were rated more than 'quite well prepared' by at least 50% of the graduates ('understanding disease processes', 'communicating effectively', 'awareness of limitations' and 'working in a team'). Similarly, more than half of educational supervisors rated graduates as more than quite competent in only three areas ('awareness of limitations', 'keeping accurate records' and 'working in a team'). Within the competencies surveyed, there were differences between the perceptions of graduates and educational supervisors on the preparedness of graduates for the skills they may require as a pre-registration house officer. CONCLUSION: Overall, given that most graduates and supervisors perceived the preparedness as 'quite well' or less, the undergraduate course had only partially met its objectives. A mismatch in ratings could be attributed to either inappropriate expectations on the part of the educational supervisors or the graduates or an inaccurate assessment by either group of respondents. PMID- 11380862 TI - Training interns in population-based research: learners' feedback from 13 consecutive batches from a medical school in India. AB - OBJECTIVES: To document learners' feedback on an educational intervention to provide interns with a hands-on learning experience in population-based research. DESIGN: Cross-sectional inquiry using a structured tool. SETTING: A medical school in India. SUBJECTS: 306 interns from 13 consecutive groups in a 3-month posting. MAIN OUTCOME MEASURES: Interns' participatory involvement in each of the nine units of learning. RESULTS: In six out of the total of nine units of learning, nearly 70% of the students rated their participatory involvement at 3 points or above (on a 5-point rating scale). This rating was 4 or 5 (good or maximal) for 50% or more students with regard to identification of research questions, review of literature, data analysis and interpretation, and for 65.4% students in data collection. However, in the last two units, on writing the research report and its presentation, a large proportion of students rated their participatory involvement as minimal or unsatisfactory (38.9% and 46.4%, respectively). For 91.2% of students, this was the first hands-on experience of any type of population-based research. When the students were asked to identify the most important factor which hampered learning during the present exposure, 54.2% of them reported that it was the unsuitable timing of the exposure, since their priority during internship was the entrance examination for postgraduate courses. CONCLUSIONS: This study empirically demonstrates that with some extra effort from teachers, interns can be exposed to a hands-on learning experience in population-based research, on a systematic basis, without additional resources. PMID- 11380863 TI - Training, job demands and mental health of pre-registration house officers. AB - OBJECTIVES: This study aims to explore pre-registration house officer (PRHO) perceptions of work role, job requirements and mental health, in order to enhance work role preparation by means of identifying potential areas for curriculum development. DESIGN: Phase 1 took place 6 weeks before completion of the pre registration year. A total of 56 PRHOs completed questionnaires which included a diary of activities, recorded daily over a 2-week period (ward rounds, on-call, audit, administration, continuing medical education, etc.), items relating to perceived occupational control, and the Maslach Burnout Inventory. In phase 2, in the final 2 weeks of the preregistration year, 36% (n=18) of phase 1 participants were randomly selected for interview. SETTING: Three hospitals in the North-west of England. RESULTS: The average working week was 56 h, with 10% working in excess of this figure. The average weekly proportion of time spent on organized continuing medical education was 5%. Routine administrative tasks took up one fifth of PRHO time and were perceived as lacking in training or educational elements. Of the PRHOs, 52% (n=29) desired further advice/training on the technical and management aspects of the job. Additional training was required on topics such as chest drains, delegation and time management. PRHOs felt this should be given prior to commencement of the pre-registration year. In terms of mental health, 25% (n=14) were experiencing burnout. Occupational control was external; many individuals perceived events as often occurring outside individual control. CONCLUSIONS: These findings have implications for the undergraduate curriculum and support General Medical Council recommendations for curriculum reform. It is suggested that organizational skills such as managing responsibility/delegation and additional training in practical procedures should be an integral part of the medical undergraduate curriculum. PMID- 11380864 TI - Extending the general practice training year: experience of one model in Scotland. AB - OBJECTIVES: These were: to describe the implementation of a scheme to extend the general practice registrar component of vocational training to 18 months in Scotland; to determine the effect of the additional attachment on registrars' confidence and skill deficits, and compare changes in these parameters with a control group of registrars who did not extend their training, and to determine whether trainers required a different training style for experienced registrars. SUBJECTS: 35 registrars extending their training, their trainers, and 39 controls who did not extend their training. SETTING: Scottish training practices. METHODS: Pre-attachment and post-attachment questionnaires for registrars, triangulated by semistructured telephone interviews, and post-attachment questionnaires for trainers. RESULTS: Extended attachments were popular with registrars and with trainers, who felt they should be a normal part of training. Registrars with extended training declared themselves to have increased in confidence and addressed skill deficits better than controls. Projects completed were assessed as being of high quality. Trainers found 10% of registrars to have a remediable, important deficit in their skills. CONCLUSION: Extended attachments appear to improve registrar confidence and to address knowledge deficits. However, only a minority of registrars had important remediable knowledge deficits and while such attachments may be desirable they may not be essential for most registrars. Overall the outcome appears to have been positive, and supports those who have argued for a change in the proportion of time spent in general practice training, but the scheme is expensive and it is difficult to set a value on what has been gained. Future schemes should have clear training objectives and be tailored towards these. PMID- 11380865 TI - Teaching sexual history taking to health care professionals in primary care. AB - OBJECTIVES: Although it is accepted that history taking is central to correct diagnosis, little work has been undertaken on the development of sexual history taking, particularly in a primary care context where sexual health may not occur to the patient. Embarrassment is recognized as one major problem. This paper reports on a series of teaching interventions designed to help primary health care professionals (doctors and nurses) to identify and deal effectively with sexual health issues in the consultation. METHODS: 141 participants took part in nine different courses, with 114 returning evaluations. All courses involved tutorial teaching on clinical and ethicolegal issues and role play with trained professional role-players; some involved video-based dramatizations to particularize principles in context. During role play sessions, which were followed by detailed, contextualized feedback, clinical issues, attitudinal issues (e.g. articulating a sense of personal embarrassment, and the risk of stereotyping), and ethicolegal issues were all discussed, as were examples of words and phrases which participants were invited to try out. OUTCOMES: The overall quality of the courses was rated by participants, on average, at 89.95 (maximum 100), and the relevance of the topic at 91.40. Free text comments centred on the power of the training as a consciousness raiser, on the need to alter communication strategies, the need to change existing clinical practice and the value of role play as a methodology. Interactive courses on sexual health are highly acceptable to participants. PMID- 11380866 TI - To BSc or not to BSc...? PMID- 11380867 TI - Speciation in rapidly diverging systems: lessons from Lake Malawi. AB - Rapid evolutionary radiations provide insight into the fundamental processes involved in species formation. Here we examine the diversification of one such group, the cichlid fishes of Lake Malawi, which have radiated from a single ancestor into more than 400 species over the past 700 000 years. The phylogenetic history of this group suggests: (i) that their divergence has proceeded in three major bursts of cladogenesis; and (ii) that different selective forces have dominated each cladogenic event. The first episode resulted in the divergence of two major lineages, the sand- and rock-dwellers, each adapted to a major benthic macrohabitat. Among the rock-dwellers, competition for trophic resources then drove a second burst of cladogenesis, which resulted in the differentiation of trophic morphology. The third episode of cladogenesis is associated with differentiation of male nuptial colouration, most likely in response to divergent sexual selection. We discuss models of speciation in relation to this observed pattern. We advocate a model, divergence with gene flow, which reconciles the disparate selective forces responsible for the diversification of this group and suggest that the nonadaptive nature of the tertiary episode has significantly contributed to the extraordinary species richness of this group. PMID- 11380868 TI - Microsatellite variation and fine-scale population structure in the wood frog (Rana sylvatica). AB - We investigated genetic population structure in wood frogs (Rana sylvatica) from a series of Prairie Pothole wetlands in the northern Great Plains. Amphibians are often thought to exist in demographic metapopulations, which require some movement between populations, yet genetic studies have revealed strong subdivision among populations, even at relatively fine scales (several km). Wood frogs are highly philopatric and studies of dispersal suggest that they may exhibit subdivision on a scale of approximately 1-2 km. We used microsatellites to examine population structure among 11 breeding assemblages separated by as little as 50 m up to approximately 5.5 km, plus one population separated from the others by 20 km. We found evidence for differentiation at the largest distances we examined and among a few neighbouring ponds, but most populations were strikingly similar in allele frequencies, suggesting high gene flow among all but the most distant populations. We hypothesize that the few significant differences among neighbouring populations at the finest scale may be a transient effect of extinction-recolonization founder events, driven by periodic drying of wetlands in this hydrologically dynamic landscape. PMID- 11380869 TI - Microsatellites reveal high levels of gene flow among populations of the California squid Loligo opalescens. AB - Information on the extent of genetic differentiation among populations of the squid Loligo opalescens is crucial for the conservation of this commercially utilized species. We analysed six highly variable microsatellite loci in 11 collections of L. opalescens from different locations and spawning seasons to estimate the relative influence of two major evolutionary forces, gene flow and genetic drift. Microsatellite allele frequency patterns suggest that gene flow prevents population differentiation in L. opalescens. Tests for genetic differentiation showed homogeneity of the samples with an overall FST/RST of 0.0028/-0.0013. Genetic uniformity among samples from different year classes indicates that allele frequency patterns in L. opalescens are relatively stable over time. However, a more complete and detailed picture of fine-scale allele frequency shifts in this species will require a systematic microsatellite analysis of local populations over consecutive spawning cycles. PMID- 11380870 TI - Breeding origin and migration pattern of dunlin (Calidris alpina) revealed by mitochondrial DNA analysis. AB - The large-scale migration of birds has been studied extensively by recoveries of ringed birds. However, there is very little ringing data from the arctic breeding grounds of waders. Here, the migration pattern of the dunlin, Calidris alpina, is studied with population genetic markers, using haplotype frequencies to estimate the breeding origin of migrating and wintering populations. Polymerase chain reaction (PCR) and restriction analysis of DNA from the mitochondrial control region was used to study the breeding origins of morphologically similar winter populations in the western Palaearctic, and to describe the population structure of the dunlin during winter. Also migrating dunlin from various stopover sites in Europe, Africa and Asia, were analysed with respect to their mitochondrial DNA (mtDNA) haplotypes. The genetic markers clearly show that the dunlin has a parallel migration system, with populations breeding in the western Palaearctic wintering mainly in the western part of the wintering range, and dunlin populations breeding further east wintering further east. The results also show that the distance between breeding and wintering area increases eastwards in this region. PMID- 11380871 TI - Correspondence between genet diversity and spatial distribution of above- and below-ground populations of the ectomycorrhizal fungus Hebeloma cylindrosporum. AB - Population studies of ectomycorrhizal fungal species have largely relied upon fruit body (the reproductive organ) sampling. Analysis of the fruit bodies alone supposes that they reflect the present and spatial organization of all below ground genets (mycorrhizas and extramatrical mycelia). The relation between fruit bodies and ectomycorrhizas was investigated for the basidiomycete agaric Hebeloma cylindrosporum in four Pinus pinaster stands in south-west France. Genet identification was based on the comparison of polymorphisms within a hypervariable segment of the ribosomal intergenic spacer amplified by polymerase chain reaction (PCR) using a H. cylindrosporum species-specific primer. Mycorrhizas were sorted from soil samples collected underneath patches of fruit bodies or patches where fruit bodies had or had not been observed during the years prior to mycorrhiza collection. On average 65% of the 1026 mycorrhizas collected underneath fruit bodies were formed by H. cylindrosporum, whereas only 2% of the 954 collected in places from where fruit bodies were absent were formed by this species. All genotypes identified above ground were also identified below ground. In patches where one genotype formed all or more than 90% of the fruit bodies, the same genotype formed all or a large majority of the mycorrhizas. In patches occupied by several different fruiting genotypes, additional nonfruiting ones could be present on the root systems. In all cases, the mycorrhizas of one genotype were found no more than 10-20 cm away from its corresponding fruit bodies, and fruit body disappearance at a given place was associated with the disappearance of the corresponding mycorrhizas within 1 year. Although there was not a strict coincidence between the total numbers of genets present below ground and of those forming fruit bodies, fruit body analysis for H. cylindrosporum appears to reflect both the genetic diversity and the spatial structure of its below-ground populations. The results obtained also illustrate the rapid turnover of ectomycorrhizal fungal species on the root systems in the absence of any obvious major disturbance of the ecosystem. PMID- 11380872 TI - Conservation genetics of peripheral populations of the mygalomorph spider Atypus affinis (Atypidae) in northern Europe. AB - Genetic diversity and differentiation were studied in peripheral populations of the recently rediscovered purse-web spider Atypus affinis in Denmark and Sweden using allozyme electrophoresis. Because of the very narrow environmental niche exploited by this species, only a limited number of potential habitats are available. Furthermore, fragmentation has reduced the number and size of potential habitats within the last century. The level of genetic diversity in A. affinis was intermediate compared with other nonsocial spiders, but low compared with invertebrates in general. Significant genetic differentiation was found within distances of only 1-10 km with FST estimates ranging from 0.020 to 0.075. Within distances of 30-60 km FST ranged from 0.081 to 0.312. Hierarchical FST revealed that genetic variability was partitioned at 89.8% within populations, 9.5% among populations within regions and only 0.7% among the four main regions in Denmark and Sweden. Comparing the result of the genetic analysis with the life history of A. affinis, it is concluded that the level of successful dispersal is low, and that the species has not recently reinvaded northern Europe but prevailed undiscovered for several decades. Finally, it is suggested that the genetic scenario found for A. affinis might represent that for a wide range of other arthropods with similar life history characteristics. PMID- 11380873 TI - Genetic diversity and structure of natural populations of Plathymenia reticulata (Mimosoideae), a tropical tree from the Brazilian Cerrado. AB - Plathymenia reticulata is a tropical tree native to the Brazilian Cerrado, one of the most important and endangered ecosystems in Brazil. This species presents high-quality wood and potential for recovery of degraded areas. Despite its importance, almost nothing is known about its genetic or ecological features. Random amplified polymorphic DNA (RAPD) markers were used to investigate the genetic diversity and structure of six natural populations of P. reticulata. DNAs from 117 adult individuals were amplified with 10 random primers and Shannon's index and amova were used to evaluate the levels of genetic diversity within and among populations. Through 72 markers, 70.8% of which were polymorphic, it was possible to obtain 117 unique RAPD phenotypes. The levels of genetic variability found in the six populations of P. reticulata were considerable and most of the genetic variation was found between individuals within populations, although pairwise PH(ST) values indicated significant divergence between populations. The among-population component accounted for, respectively, 12.3% and 16% of the genetic variation, according to amova and Shannon's index. These results were compared with other genetic studies on plant species and such a level of differentiation among populations corresponds to that which has usually been observed for outcrossing plants. The importance of maintenance of the P. reticulata populations and implications of the analysis of adult individuals, considering the longevity of this species and the relatively recent Cerrado fragmentation, are discussed. PMID- 11380874 TI - Fragmentation of riverine systems: the genetic effects of dams on bull trout (Salvelinus confluentus) in the Clark Fork River system. AB - Migratory bull trout (Salvelinus confluentus) historically spawned in tributaries of the Clark Fork River, Montana and inhabited Lake Pend Oreille as subadult and adult fish. However, in 1952 Cabinet Gorge Dam was constructed without fish passage facilities disrupting the connectivity of this system. Since the construction of this dam, bull trout populations in upstream tributaries have been in decline. Each year adult bull trout return to the base of Cabinet Gorge Dam when most migratory bull trout begin their spawning migration. However, the origin of these fish is uncertain. We used eight microsatellite loci to compare bull trout collected at the base of Cabinet Gorge Dam to fish sampled from both above and further downstream from the dam. Our data indicate that Cabinet Gorge bull trout are most likely individuals that hatched in above-dam tributaries, reared in Lake Pend Oreille, and could not return to their natal tributaries to spawn. This suggests that the risk of outbreeding depression associated with passing adults over dams in the Clark Fork system is minimal compared to the potential genetic and demographic benefits to populations located above the dams. PMID- 11380875 TI - Mode of reproduction and amplified fragment length polymorphism variation in purple needlegrass (Nassella pulchra): utilization of natural germplasm sources. AB - A dominant plant of the California grasslands, purple needlegrass [Nassella pulchra (Hitchc.) Barkworth] is an important revegetation species in its native range. The amplified fragment length polymorphism (AFLP) method was used to elucidate mode of reproduction and nucleotide variation among 11 natural populations and three selected natural germplasm releases of N. pulchra. A total of 12 co-dominant AFLPs, informative within eight populations, failed to reveal any heterozygous individuals, indicating very high selfing rates (S(H)=1). Estimates of nucleotide diversity within populations ranged from 0 to 0.00069 (0.00035 average), whereas the total nucleotide divergence among populations ranged from 0.00107 to 0.00382 (0.00247 average). Measures of population differentiation (GS) in terms of Shannon-Weaver diversity values and estimated nucleotide substitutions were 0.90 and 0.86, respectively. Although some of the sample populations contained a mixture of true breeding genotypes, most populations could be distinguished unambiguously. Moreover, geographical distance between the natural source populations was significantly correlated with genetic distance (r = 0.60) among the corresponding sample populations. Results indicate that inbreeding, combined with founder effects and/or selection, has contributed to the differentiation of N. pulchra populations. Foundation seed populations of the selected natural germplasm releases were genetically well defined and most similar to natural seed collected near the corresponding source populations. Thus, these commercial germplasm sources will be made practically available and useful for conservation plantings within the intended areas of utilization. PMID- 11380876 TI - Reconciling patterns of inter-ocean molecular variance from four classes of molecular markers in blue marlin (Makaira nigricans). AB - Different classes of molecular markers occasionally yield discordant views of population structure within a species. Here, we examine the distribution of molecular variance from 14 polymorphic loci comprising four classes of molecular markers within approximately 400 blue marlin individuals (Makaira nigricans). Samples were collected from the Atlantic and Pacific Oceans over 5 years. Data from five hypervariable tetranucleotide microsatellite loci and restriction fragment length polymorphism (RFLP) analysis of whole molecule mitochondrial DNA (mtDNA) were reported and compared with previous analyses of allozyme and single copy nuclear DNA (scnDNA) loci. Temporal variance in allele frequencies was nonsignificant in nearly all cases. Mitochondrial and microsatellite loci revealed striking phylogeographic partitioning among Atlantic and Pacific Ocean samples. A large cluster of alleles was present almost exclusively in Atlantic individuals at one microsatellite locus and for mtDNA, suggesting that, if gene flow occurs, it is likely to be unidirectional from Pacific to Atlantic oceans. Mitochondrial DNA inter-ocean divergence (FST) was almost four times greater than microsatellite or combined nuclear divergences including allozyme and scnDNA markers. Estimates of Neu varied by five orders of magnitude among marker classes. Using mathematical and computer simulation approaches, we show that substantially different distributions of FST are expected from marker classes that differ in mode of inheritance and rate of mutation, without influence of natural selection or sex-biased dispersal. Furthermore, divergent FST values can be reconciled by quantifying the balance between genetic drift, mutation and migration. These results illustrate the usefulness of a mitochondrial analysis of population history, and relative precision of nuclear estimates of gene flow based on a mean of several loci. PMID- 11380877 TI - Microsatellite variation in natural Drosophila melanogaster populations from New South Wales (Australia) and Tasmania. AB - Microsatellite variation was studied at 48 microsatellite loci in 10 Drosophila melanogaster populations to investigate the population structure on the Australian east coast. Low, but statistically significant population differentiation was observed among most populations. The populations on the Australian mainland did not show evidence for isolation by distance. We conclude that the population structure of D. melanogaster on the Australian mainland is probably the result of a shared history (recent colonization). The observed differences between local D. melanogaster populations probably reflect variation in effective population sizes rather than patterns of gene flow. Two populations from Tasmania were more differentiated from the Australian mainland than a population from Israel, raising the question whether they are derived from the Australian mainland or colonized from a different source population. PMID- 11380878 TI - Population structure in two sympatric species of the Lake Tanganyika cichlid tribe Eretmodini: evidence for introgression. AB - Patterns of genetic differentiation were analysed and compared in two sympatric species of the endemic Lake Tanganyika cichlid tribe Eretmodini by means of mitochondrial DNA (mtDNA) sequences of the control region and six microsatellite DNA loci. The sample area covers a total of 138 km of mostly uninterrupted rocky shoreline in the Democratic Republic of Congo and includes the entire distribution range of Tanganicodus cf. irsacae that stretches over a distance of 35 km. Both markers detected significant genetic differentiation within and between the two species. T. cf. irsacae contained lower overall genetic variation than Eretmoduscyanostictus, possibly due to its more restricted range of distribution and its smaller effective population sizes. Complete fixation of Tanganicodus mtDNA haplotypes was observed in Eretmodus at two localities, while at two other localities some Tanganicodus individuals possessed Eretmodus mtDNA haplotypes. Taking into account the relatively large average sequence divergence of 6.2% between the two species, as well as the geographical distribution of mtDNA haplotypes in the lake, the observed pattern is more likely to be a consequence of asymmetric introgression than of shared ancestral polymorphism. As there is significant population differentiation between sympatric Tanganicodus and Eretmodus populations, the events of introgressions may have happened after secondary contact, but our data provide no evidence for ongoing gene flow and suggest that both species are reproductively isolated at present time. PMID- 11380879 TI - Phylogeography of the dusky shrew, Sorex monticolus (Insectivora, Soricidae): insight into deep and shallow history in northwestern North America. AB - Phylogenetic relationships among the dusky shrew (Sorexmonticolus) and eight related species (S. bairdi, S. bendirii, S. neomexicanus, S.ornatus, S. pacificus, S. palustris, S. sonomae and S.vagrans) were assessed using sequences from the mitochondrial cytochrome b gene (801 bp). Analyses using parsimony and maximum likelihood revealed significant molecular variation not reflected in previous morphological studies of these species. Conversely, three morphologically defined species (S.bairdi, S.neomexicanus and S.pacificus) were poorly differentiated. Sorexornatus and S.vagrans represented basal taxa for a more inclusive group that included: (i) a widespread Continental clade containing S.monticolus (Arizona to Alaska, including S. neomexicanus); (ii) a Coastal clade containing S.monticolus (Oregon to south-east Alaska, including S. bairdi and S. pacificus); (iii) the semiaquatic species (S. bendirii and S. palustris); and (iv) S.sonomae. Additional subdivision was observed within the Continental clade corresponding to populations from the northern and southern Rocky Mountains. Average uncorrected sequence divergence between the Coastal and Continental clades was 5.3% (range 4.5-6.2%), which exceeds many interspecific comparisons within this species complex and within the genus Sorex. Lack of resolution of internal nodes within topologies suggests a deep history of rapid diversification within this group. Late Pleistocene/Holocene glacial perturbations are reflected in the shallow phylogeographic structure within these clades in western North America. Our results suggest also that S. monticolus is not monophyletic under current taxonomic nomenclature. This perspective on phylogeographic history was developed within a growing comparative framework for other organisms in western North America. PMID- 11380880 TI - Haplotype (mtDNA) diversity of brown trout Salmo trutta in tributaries of the Austrian Danube: massive introgression of Atlantic basin fish--by man or nature? AB - Mitochondrial haplotype diversity in 27 populations of brown trout, Salmo trutta L., in Austria was investigated by sequencing the 5' end of the mitochondrial DNA (mtDNA) control region. Although all populations are within the Danube drainage, 44% of all individuals carried Atlantic basin haplotypes. It is argued that the presence of these haplotypes in Austria primarily reflects introgression stemming from the stocking of hatchery-reared fish. However, several lines of evidence suggest that some natural colonization from Atlantic lineages may have contributed to the present haplotype diversity. Nonetheless, the more diverse Danubian clade is represented by regionally distinct haplotype diversity that should be protected from the continued introduction of domesticated strains of exogenous fish PMID- 11380881 TI - Phylogeography of the yucca moth Tegeticula maculata: the role of historical biogeography in reconciling high genetic structure with limited speciation. AB - Tegeticula maculata is one of the most ancient and morphologically variable lineages within the yucca moths, yet has apparently undergone little diversification in comparison with much younger yucca moth lineages that have rapidly diversified. A phylogeographic approach was used to determine the number of independent lineages within T. maculata and to examine whether these patterns corresponded with morphological differences between its subspecies maculata and extranea. Phylogenetic analysis of mitochondrial DNA sequence variation indicated that the two subspecies are in separate clades, but there was also an equally deep split within subspecies maculata. There was no evidence for gene flow among regions and there was considerable substructure within clades. The phylogeographic structure of moth populations among and within subspecies can be explained in part by historical biogeographic boundaries and increasingly patchy postglacial distribution of the exclusive host plant, Hesperoyucca whipplei. Local specialization and co-adaptation would be possible in the absence of apparent gene flow, yet gross morphological divergence is limited to the very old split between the subspecies. Sorting of ancient mitochondrial lineages followed by local genetic differentiation may explain the pattern of high genetic structure with limited speciation. PMID- 11380882 TI - Haplotype variation in a mitochondrial tandem repeat of Norway spruce (Picea abies) populations suggests a serious founder effect during postglacial re colonization of the western Alps. AB - Populations from 13 elevational transects of Norway spruce [Picea abies (L.) Karst] across the Alpine range were sampled to elucidate the geographical pattern of genetic variation in relation to postglacial re-colonization and to study elevational effects on haplotypic diversity. We assessed fragment length variation in a tandem repeat region of the mitochondrial (mt) nad1 intron 2. This maternally inherited genetic marker is suited to infer migration as it is dispersed by seed only. A total of 10 haplotypes was found, most of which were due to repeat copy number variation. An analysis of molecular variance (amova) showed that overall population differentiation was high (F(ST)=0.41), and it revealed a significant differentiation between monomorphic western and moderately to highly variable eastern Alpine populations. This phylogeographic pattern may be explained by a founder effect during postglacial re-colonization. An early arriving haplotype, assumed to originate from a western Carpathian refugium, could expand into suitable habitats, reducing the chances for establishment of subsequently arriving haplotypes. On the other hand, the high variation in populations within an Italian transect of the south-eastern Alps may be the consequence of merging migration pathways from and close distance to putative glacial refugia, most likely those assumed in the Carpathian mountains and on the Balkan peninsula or possibly in the central plains of Italy. An effect of elevation on haplotypic diversity was not evident, though a low, but significant, partition of total genetic variation was attributed to among-population variation in one Italian transect. Various factors, such as vertical seed dispersal and forest management, may account for blurring an otherwise established pattern of genetic variation on a small geographical scale. PMID- 11380883 TI - Assignment of paternity groups without access to parental genotypes: multiple mating and developmental plasticity in squid. AB - We present a novel approach to investigating sibling relationships and reconstructing parental genotypes from a progeny array. The Bayesian method we have employed is flexible and may be applicable to a variety of situations in addition to the one presented here. While mutation rates and breeding population allele frequencies can be taken into account, the model requires relatively few loci and makes few assumptions. Paternity of 270 veined squid (Loligo forbesi) hatchlings from three egg strings collected from one location was assigned using five microsatellite loci. Paternal and maternal genotypes reconstructed for each of the three strings were identical, strongly indicating that a single female produced the strings that were fertilized by the same four males. The proportion of eggs fertilized was not equal between males in all three strings, with male 1 siring most offspring (up to 68% in string 1), through to male 4 siring the least (as low as 2.4% in string 1). Although temperature had a profound effect on incubation time, paternity did not affect this trait at 12 degrees C or 8 degrees C. PMID- 11380884 TI - Noninvasive paternity assignment in Gombe chimpanzees. AB - The relative success of chimpanzee male mating strategies, the role of male dominance rank and the success of inbreeding avoidance behaviour can only be assessed when paternities are known. We report the probable paternities of 14 chimpanzees included in a long-term behavioural study of chimpanzees (Pan troglodytes schweinfurthii) at Gombe National Park, Tanzania. DNA samples were collected noninvasively from shed hair and faeces and genotyped using 13-16 microsatellite loci characterized in humans. All 14 offspring could be assigned to fathers within the community. While there is a positive relationship between male rank and reproductive success, we demonstrate that a range of male mating strategies (possessiveness, opportunistic mating and consortships) can lead to paternity across all male ranks. Several adult females were at risk of breeding with close male relatives. Most successfully avoided close inbreeding but in one case a high-ranking male in the community mated with his mother and produced an offspring. In contrast to recent data on chimpanzees (P. t. verus) from the Tai forest, Cote d'Ivoire, no evidence of extra-group paternity was observed in our study. Reanalysis of Tai data using a likelihood approach casts doubt on the occurrence of extra-group paternity in that community as well. PMID- 11380885 TI - High levels of extra-pair paternity in an isolated, low-density, island population of tree swallows (Tachycineta bicolor). AB - Molecular genetic studies have suggested that apparently nonbreeding males ('floaters') may account for a significant proportion of extra-pair paternity (EPP) in avian populations. Attempts to determine the influence of breeding density on EPP are therefore confounded by the presence of a subpopulation of floaters whose numbers are difficult to estimate. To study EPP in a tree swallow (Tachycineta bicolor) population with few floaters, we chose a nestbox grid on an island with an excess of available breeding sites and very few floaters. We obtained DNA samples from 13 complete families and performed DNA profiling on them using four microsatellite loci. For comparison, we also obtained a sample of 58 extra-pair young (EPY) from a mainland population typed at five microsatellite loci. Paternity assignments among resident males in both populations were made using the microsatellite profiles and a likelihood-based statistical method. Of the 67 island nestlings typed, we found 37 (55%) nestlings from 11 (85%) different nests that were EPY. The proportion of nestlings that were EPY and the proportion of broods containing EPY did not differ significantly between island and mainland populations studied previously. There was no significant difference between island and mainland populations in the proportion of extra-pair paternities assigned among neighbouring resident males. Male breeding density does not appear to affect the ability of female tree swallows to obtain extra pair fertilizations, at least over the range of densities studied so far. The rate of EPP has remained remarkably consistent over many years, studies and populations implying an important role of active female choice in determining EPP. PMID- 11380886 TI - Mating patterns, relatedness and the basis of natal philopatry in the brown long eared bat, Plecotus auritus. AB - The brown long-eared bat, Plecotus auritus, is unusual among temperate zone bats in that summer maternity colonies are composed of adult males and females, with both sexes displaying natal philopatry and long-term association with a colony. Here, we describe the use of microsatellite analysis to investigate colony relatedness and mating patterns, with the aim of identifying the evolutionary determinants of social organization in P. auritus. Mean colony relatedness was found to be low (R=0.033 +/- 0.002), with pairwise estimates of R within colonies ranging from -0.4 to 0.9. The proportion of young fathered by males in their own colony was investigated using a Bayesian approach, incorporating parameters detailing the number of untyped individuals. This analysis revealed that most offspring were fathered by males originating from a different colony to their own. In addition, we determined that the number of paternal half-sibs among cohorts of young was low, inferring little or no skew in male reproductive success. The results of this study suggest that kin selection cannot account for colony stability and natal philopatry in P. auritus, which may instead be explained by advantages accrued through the use of familiar and successful roost sites, and through long-term associations with conspecifics. Moreover, because the underlying causes of male natal dispersal in mammals, such as risk of inbreeding or competition for mates, appear to be avoided via extra-colony copulation and low male reproductive skew, both P. auritus males and females are able to benefit from long-term association with the natal colony. PMID- 11380887 TI - Demic structure and its relation with the distribution of an adaptive trait in Danish flea beetles. AB - The flea beetle Phyllotreta nemorum is an oligophagous species using crucifers as host plants. In Denmark two populations have been found which use Barbarea vulgaris ssp. arcuata (G-type) as a host plant, whereas this plant is unsuitable for the survival of the majority of P. nemorum. In the locations in which these two populations occur, alternative host plants are also present. The plants occur in patches, some of which contain a mixture of host plants. In this study of allozyme variation, genetic differentiation between P. nemorum using different host plants in patches in the two different localities was studied hierarchically to assess substructuring of the populations. Evidence was found for low, but significant, amounts of genetic differentiation between (sub)populations using spatially separated plant patches at a distance of approximately 100 m to 1 km (theta = 0.009) and between localities approximately 44 km apart (theta = 0.026), and there was an association between genetic differentiation and geographical distance. No genetic differentiation was found between beetles from different host plants with overlapping local distributions. No evidence was thus found for sympatric host race formation. The geographical distribution of genes enabling P. nemorum to use B. vulgaris as a host plant (100% 'resistant' beetles in samples from B. vulgaris, but much fewer on patches containing only alternative host plants) contrasts with the relatively low amount of genetic differentiation at the neutral allozyme loci. This distribution of 'resistant' beetles (to B. vulgaris defence) is likely to be influenced by local differences in selection and asymmetric gene flow. PMID- 11380888 TI - Lack of molluscan host diversity and the transmission of an emerging parasitic disease in Bolivia. AB - Fasciolosis is a re-emerging parasitic disease that affects an increasing number of people in developing countries. The most severe endemic affects the Bolivian Altiplano, where the liver fluke (Fasciola hepatica) and its hermaphroditic snail host, Lymnaea truncatula, have been introduced from Europe. To achieve a better understanding of the epidemiological situation and the consequences of the colonization event of this invasive species, genetic analysis of Bolivian snail populations was needed. Here we compare the genetic diversity and population structure of snail samples from the Bolivian Altiplano with samples from the Old World at six polymorphic microsatellite loci. Whereas some variability exists in the snail populations from the Old World, we observe only a single genotype of L. truncatula in the Bolivian Altiplano. We discuss the possible explanations for such a reduction in genetic variability, and, given the high natural parasitism pressures exerted on the snail populations, we discuss the relevance of this result for host-parasite interactions. PMID- 11380889 TI - The 'rare allele phenomenon' in a ribosomal spacer. AB - We describe the increased frequency of a particular length variant of the internal transcribed spacer 1 (ITS-1) of the ribosomal DNA in a hybrid zone of the land snail Albinaria hippolyti. The phenomenon that normally rare alleles or other markers can increase in frequency in the centre of hybrid zones is not new. Under the term 'hybrizyme' or 'rare allele' phenomenon it has been recorded in many organisms and different genetic markers. However, this is the first time that it has been found in a multicopy locus. On the one hand, the pattern fits well with the view that purifying selection in hybrid populations works on many loci across the genome and should thus have its effect on many independent molecular markers. On the other hand, the results are puzzling, given that the multiple copies of rDNA are not expected to respond in unison. We suggest two possible explanations for these conflicting observations. PMID- 11380890 TI - Food allergy: immunology of the GI mucosa towards classification and understanding of GI hypersensitivities. PMID- 11380891 TI - Structure of food allergens in relation to allergenicity. PMID- 11380893 TI - Early sensitisation to food antigens--when and how? PMID- 11380892 TI - The immunological role of breast feeding. PMID- 11380894 TI - Diagnosis of food allergy: tests in vivo and in vitro. PMID- 11380895 TI - Diagnosis of food allergy: the oral provocation test. PMID- 11380896 TI - The role of the atopy patch test (APT) in diagnosis of food allergy in infants and children with atopic dermatitis. PMID- 11380897 TI - GI manifestations of food allergy. PMID- 11380898 TI - Gastrointestinal allergy: clinical symptoms and immunological mechanisms. PMID- 11380899 TI - Clinical relevance of villous atrophy. PMID- 11380900 TI - The gastrointestinal flora and the skin--Is there a link? PMID- 11380901 TI - Probiotics in the prevention and treatment of allergic disease. PMID- 11380902 TI - Atopic dermatitis in infancy and childhood: an ongoing challenge. PMID- 11380903 TI - Atopic dermatitis: immunological mechanisms in relation to phenotype. PMID- 11380904 TI - Clinical features and diagnostic criteria of atopic dermatitis in relation to age. PMID- 11380905 TI - Aspects of nutritional management of food allergy. PMID- 11380906 TI - Primary and secondary dietary prevention. PMID- 11380907 TI - Soy and other protein sources. PMID- 11380908 TI - Immunological approaches to the treatment of food allergy. PMID- 11380909 TI - Food-induced anaphylaxis. PMID- 11380910 TI - Future aspects of pharmacological treatment to inhibit the allergic march. PMID- 11380911 TI - Re-treatment for immune globulin-resistant Kawasaki disease: a comparative study of additional immune globulin and steroid pulse therapy. AB - BACKGROUND: We compared the efficacy and safety of additional intravenous immune globulin (IVIG) therapy with steroid pulse therapy in patients with IVIG resistant Kawasaki disease. METHODS: Two-hundred and sixty-two consecutive patients had been treated with a single dose of IVIG (2 g/kg) and aspirin (30 mg/kg per day). Thirty-five patients (13.4%) were not clinical responders to the initial IVIG treatment. They received an additional IVIG treatment (1 g/kg) within 48 h after the initial treatment. Seventeen patients (6.5%) did not respond to the additional IVIG treatment. We randomly divided these patients into two groups: group 1 consisted of eight patients who were treated with a single additional dose of IVIG (1 g/kg), while group 2 consisted of nine patients who were treated with steroid pulse therapy. RESULTS: The IVIG-resistant patients had a high incidence of coronary artery lesions (CAL; 48.6%). Five patients (62.5%) in group 1 had CAL, including two patients who each had a giant aneurysm and three patients who each had a small aneurysm. Seven patients (77.8%) in group 2 had CAL, including two patients who each had a giant aneurysm, two patients who each had a small coronary aneurysm and three patients who each showed transient dilatation during steroid pulse therapy. There was no significant difference in the incidence of CAL between the two groups. The duration of high fever in group 2 (1.4~0.7 days) was significantly shorter than in group 1 (4.8~3.4 days; P<0.05). The medical costs for the treatment of patients in group 2 (113, 012 yen +/- 22,084) were significantly lower than those for group 1 (144,194 yen +/- 12,914; P<0.05). CONCLUSIONS: Steroid pulse therapy may be useful in the treatment of patients with IVIG-resistant Kawasaki disease who experience prolonged fever. However, transient dilatation of the coronary artery is observed during steroid pulse therapy, so careful echocardiographic examination should be performed for those patients receiving steroid pulse therapy for the sake of early detection of coronary artery abnormalities. PMID- 11380912 TI - Dobutamine stress surface mapping of myocardial ischemia in Kawasaki disease. AB - BACKGROUND: To detect and localize myocardial ischemia, a method that does not require physical exertion is sometimes needed in children with Kawasaki disease. METHODS: Dobutamine stress body surface mapping was performed in 115 children with a history of Kawasaki disease (58 without coronary artery lesions, 40 who had coronary lesions without myocardial ischemia and 17 with myocardial ischemia). The maximum infusion rate of dobutamine was 30 microg/kg per min. Myocardial ischemia was diagnosed by the presence of an area of hypoperfusion on scintigraphy at rest and/or an increase in hypoperfusion during a dobutamine stress test compared with resting scintigraphy. We studied the number of leads that showed significant ST depression on the isopotential map (nST), the number of the row containing the lead with the smallest negative value on the isointegral map (I(min)), and the localization of myocardial ischemia on the isointegral map. Based on findings in patients without coronary artery lesions, we defined the criteria for detecting myocardial ischemia as nST > or = 1 and I(min) < or = 4. RESULTS: The sensitivity of detecting myocardial ischemia was 94.1% using nST and 41.7% using I(min), while the specificity of these methods was 98.9 and 96.9%, respectively. The localization of myocardial ischemia on stress body surface mapping was 100% concordant with that determined by stress myocardial scintigraphy. CONCLUSIONS: Dobutamine stress body surface mapping for the detection of myocardial ischemia is a non-invasive, more convenient and repeatable test compared with exercise myocardial scintigraphy and it is a more objective test compared with exercise echocardiography. Dobutamine stress body surface mapping is useful for the identification and localization of silent myocardial ischemia in pediatric patients with Kawasaki disease, especially those who cannot perform tests involving physical exercise. PMID- 11380913 TI - Effect of dexamethasone therapy on pulmonary function in chronic lung disease: a comparison of disease types. AB - BACKGROUND: In the present study, we investigated the effect of dexamethasone (DEX) therapy on extubation and pulmonary function in patients with chronic lung disease (CLD) who required long-term mechanical ventilation. In addition, we compared the effects of DEX therapy among CLD types. METHODS: Twenty-two CLD patients who were ventilator dependent for 28 days or longer received DEX therapy for the purposes of extubation. A tapering dose of DEX, starting from 0.5 mg/kg per day, was administered for 7 days. Pulmonary function was measured at initiation of administration and 4 days after initiation. We evaluated static respiratory system compliance (Crs) and static respiratory system resistance (Rrs) adjusted by bodyweight. Chronic lung disease types were categorized according to the classification of the Ministry of Health and Welfare Research Project. We compared the effect of DEX therapy among CLD types. RESULTS: Dexamethasone therapy was started at a mean (+/-SD) 45 =/- 11 days after birth and 32.1 +/- 1.3 weeks of postconceptional age in infants with a mean bodyweight of 939 +/- 153 g. After DEX therapy, extubation was successful in all 22 patients. Following DEX administration, Crs was significantly increased from 0.69 +/- 0.13 to 1.17 +/- 0.21 mL/cm H2O per kg. In contrast, Rrs did not show any clear changes. Comparing CLD types, no difference was observed for Crs and Rrs in each disease type. CONCLUSIONS: Dexamethasone was administered to CLD patients requiring long-term mechanical ventilation for the purposes of extubation and extubation was successful in all patients. It was found that Crs was increased in all patients following DEX, regardless of CLD type. The increase in Crs following DEX administration may have been related to successful extubation. PMID- 11380914 TI - Hypercalciuria in children with febrile convulsions. AB - BACKGROUND: The purpose of the present study was to investigate whether idiopathic hypercalciuria may be implicated in the pathogenesis of febrile convulsions. METHODS: We studied 38 children (22 boys) with febrile convulsions (mean (+/- SD) age 3.25 +/- 1.09 years) and 45 healthy children (28 boys) of similar age who served as controls. Twenty-four hour urine calcium and phosphate, as well as serum calcium, phosphate, alkaline phosphatase and intact parathyroid hormone (PTH) concentrations were determined. RESULTS: Hypercalciuria (urine Ca >4.0 mg/kg bodyweight per 24 h) was found in nine children with febrile convulsions (23.7%) and in three controls (6.7%). Hypercalciuric children excreted significantly more phosphate in their urine (37.0 +/- 11.6 mg/kg bodyweight per 24 h) than normocalciuric children (18.7 +/- 8.7 mg/kg bodyweight per 24 h) and controls (20.2 +/- 7.6 mg/kg bodyweight per 24 h). They also had higher serum intact PTH concentrations (49.87 +/- 15.36 pg/mL) than normocalciuric (35.39 +/- 15.67 pg/mL) and control children (28.21 +/- 14.00 pg/mL). According to the calcium-loading test, eight of nine children with hypercalciuria had the renal type of the disorder. Furthermore, hypercalciuric children had significantly more convulsive episodes (2.77 +/- 1.98) than normocalciuric children (1.86 +/- 1.24). CONCLUSIONS: Our results suggest that renal hypercalciuria may be implicated in the pathogenesis of febrile convulsions. PMID- 11380915 TI - Proinflammatory cytokines, prostaglandins and zinc in febrile convulsions. AB - BACKGROUND: Some changes in the levels of proinflammatory cytokines, prostaglandins and zinc (Zn) in peripheral blood and cerebrospinal fluid (CSF) have been suggested to occur for the pathogenesis of febrile convulsions (FC). METHODS: In order to test this hypothesis, the levels of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 alpha, IL-1 beta and prostaglandins (PGE(2), PGF(2 alpha), PGD(2)) in the CSF and plasma and the levels of Zn in serum and CSF were investigated in children during the acute and late phases of FC. Results were compared with control subjects with meningismus. RESULTS: During the acute phase of FC, children had significantly elevated plasma levels of IL-1 beta, CSF levels of TNF-alpha, plasma levels of PGE(2), PGF(2 alpha) and PGD(2) and CSF levels of PGD(2) (P<0.05). A positive correlation between the degree of fever and plasma IL-1 beta levels was observed in both patients and controls. Three months after the acute phase of FC, plasma levels of IL-1 beta had returned to levels seen in controls. Children with FC also had significantly decreased serum Zn levels during the acute phase (P<0.05). However, there was no significant difference between the groups with respect to CSF Zn levels (P>0.05). CONCLUSIONS: During the acute phase of FC, patients had significantly increased plasma IL-1 beta and prostaglandin levels and decreased serum Zn levels. These changes may be responsible for FC pathogenesis. PMID- 11380916 TI - Correlation between neuroimaging and neurological outcome in periventricular leukomalacia: diagnostic criteria. AB - BACKGROUND: Periventricular leukomalacia (PVL) is the most important factor in cerebral palsy in preterm infants. METHODS: In the present study, we investigated 747 preterm infants of less than 36 weeks gestation who were repeatedly examined by cranial ultrasonography and computed tomography (CT) scanning at around 40 weeks of corrected post-menstrual age. The clinical course of these infants was followed for more than 3 years and they were examined by magnetic resonance imaging (MRI) between 12 and 18 months of age. RESULTS: Single examinations in early infancy were not sufficient to diagnose PVL, but the combination of ultrasonography, CT and MRI examinations allowed the clinical diagnosis of PVL. In preterm infants, clinical PVL could be predicted from cystic PVL and periventricular echogenicity (PVE) 3 or PVE 2 prolonged over 3 weeks on ultrasonography and confirmed by MRI after 11 months of corrected age. CONCLUSIONS: We tried to determine diagnostic criteria for PVL by neuroimaging. Such criteria from neuroimaging for PVL may be useful for determining the exact occurrence rate of and clinical risk factors for PVL. PMID- 11380917 TI - Impact of glycemic control on serum lipoprotein (a) in Arab children with type 1 diabetes. AB - BACKGROUND: Lipoprotein (a) (Lp (a)) is an independent risk factor for coronary artery disease (CAD), a major cause of death in patients with type 1 diabetes mellitus. Both type 1 diabetes and CAD represent major problems in Kuwait. Data on the effect of metabolic control on Lp (a) in diabetic children are limited and this is particularly true for Arab children. The objectives of the present study were to analyze serum Lp (a) levels in patients with type 1 diabetes compared with non-diabetic children, taking into account the effect of glycemic control. METHODS: Circulating lipids, including Lp (a), were measured in serum samples from 60 prepubertal non-diabetic children and 58 prepubertal children with type 1 diabetes. Comparisons of Lp (a) concentrations were made between the non-diabetic and diabetic children with good to fair control (glycosylated hemoglobin (GHb) <11%) and a group of diabetic children with poor control (GHb > or = 11%). RESULTS: The mean serum Lp (a) level in all diabetic children was 187.62+160.43 mg/L, compared with 162.88+156.06 mg/L in the control group. The group of children with poor glycemic control had higher median Lp (a) levels (147.50 mg/L) than either the group of diabetic children with good to fair control (95 mg/L; P<0.028) or the group of non-diabetic children (125 mg/L; P<0.04). Moreover, 38.3% of poorly controlled diabetic children had elevated Lp (a) levels > or = 250 mg/L, compared with 12.5% of diabetic children with good to fair control and 16.7% of non-diabetic children (P<0.025 and P<0.039, respectively). No association was found between Lp (a), diabetes duration and insulin dose. CONCLUSIONS: In Arab children, highest Lp (a) levels are associated with poorest metabolic control. The prevalence of Lp (a) levels associated with cardiovascular risk is higher in poorly controlled diabetic children. Increased levels of Lp (a) may be another contributing factor to the high risk for CAD in diabetic patients. PMID- 11380918 TI - Development of sucking behavior in infants who have not been fed for 2 months after birth. AB - BACKGROUND: Term infants are able to develop sucking behavior after birth. However, the requirements for this development have not yet been fully elucidated. In the present study, we investigated whether an oral-feed practice is necessary for the development of sucking behavior and whether non-nutritive sucking could support the development of this behavior while the infants cannot be fed. METHODS: Subjects of the present study were four term or near-term infants who had never been fed orally for approximately 2 months after birth because of gastrointestinal problems. Sucking pressure was measured with a silicone tube inserted into an artificial nipple and sucking efficiency was calculated during an entire feeding. RESULTS: Sucking pressure, frequency and duration at the first time oral feed corresponded to values obtained for normal term infants at their first oral feed. Although infants who had not received oral feeding demonstrated poor sucking abilities initially, sucking performance improved with practice. Sucking parameters measured 1 month since oral feeding was initiated in these infants corresponded to values obtained for normal-term infants at 1 month of age. Although infants who had not received oral feeding had often sucked a pacifier for sedation, this non-nutritive sucking did not result in the development of sucking behavior. CONCLUSIONS: These results demonstrate that an oral feeding practice is necessary for the development of sucking behavior and that non-nutritive sucking does not affect the development of sucking. PMID- 11380919 TI - Ala/Thr60 variant of the Leydig insulin-like hormone is not associated with cryptorchidism in the Japanese population. AB - BACKGROUND: Leydig insulin-like hormone (Insl3), a member of the insulin-like superfamily, is specifically expressed in Leydig cells of fetal and postnatal murine testis. Recently, the absence of the Insl3 gene has been reported to result in bilateral cryptorchidism in male mice and it has been suggested that mutations of the INSL3 gene may cause cryptorchidism in humans. METHODS: We sequenced the INSL3 gene from five Japanese patients with sporadic bilateral cryptorchidism. Patients' genome DNA was prepared from blood leukocytes. Two exons of the INSL3 gene were amplified by polymerase chain reaction and were sequenced directly. A restriction fragment length polymorphism assay was performed on 70 control samples for analysis of polymorphism. RESULTS: Three of five cases had a heterozygous single-base change, a G to A transition at position 178 of the INSL3 gene, which predicts an alanine (GCC) to threonine (ACC) change at codon 60 (designated A60T). However, the A60T mutation was also found in the normal Japanese population at an allele frequency of 26%, which suggests that this mutation is a common polymorphism and is not associated with the occurrence of cryptorchidism. CONCLUSIONS: No mutation has been found in the INSL3 gene from Japanese patients with idiopathic cryptorchidism. We did find the A60T polymorphism, which was not associated with the occurrence of cryptorchidism. PMID- 11380920 TI - Fiberoptic colonoscopic polypectomy in childhood: report and review of cases. AB - BACKGROUND: Fiberoptic colonoscopy has been a routine therapeutic modality for colorectal polyps in pediatric patients. Methods of bowel preparation, anesthesia, area of investigation and treatment depending on histopathology are still controversial. In order to clarify the rationale of pediatric colonoscopy the present study was performed. METHODS AND RESULTS: We analyzed the results of colonoscopic examination in 21 patients with colorectal polyps. Mean patient age was 3.7 years, with a range of 1--7 years. Rectal polyps were seen in 10 cases: seven had a solitary polyp (juvenile in six and adenoma in one) and three had multiple polyps (juvenile, lymphoid and Peutz--Jeghers coexisting with hyperplastic polyps). Sigmoid colon polyps were seen in 10 cases: all were solitary juvenile polyps, but one had adenomatous change. Another had multiple Peutz-- Jeghers polyps located in the entire colon. Flexible colonoscopic polypectomy was performed in 16 patients and transanal polypectomy was performed in four patients. Autoamputation was seen in two cases of juvenile polyp (resection was ultimately performed in a case having repeated autoamputation). After removing the polyps, all patients have had no recurrence for a period ranging from 6 months to 15 years, except for one case with Peutz--Jeghers syndrome. CONCLUSIONS: Most polyps are located in the rectum or the sigmoid colon. Although the majority are solitary or juvenile polyps, because histopathologic variety is seen in pediatric colon polyps, histopathologic examination of each polyp is important to detect any dysplastic or adenomatous element with malignant potential and to make a suitable follow-up schedule. Symptomatic polyps should be removed by fiberoptic colonoscopy or transanal resection with total colon endoscopic examination under general anesthesia. Polypectomy using the electrocautery snare and clip is effective and safe and bowel preparation using polyethylene glycol electrolyte solution is sufficient for the procedure. PMID- 11380921 TI - Mutation analysis of Crouzon syndrome and identification of one novel mutation in Taiwanese patients. AB - BACKGROUND: Crouzon syndrome is an autosomal dominant disorder causing premature fusion of the cranial suture. Mutations have been reported in exon IIIa or IIIc of the fibroblast growth factor receptor 2 (FGFR2) gene. METHODS: In the present study, nine unrelated Crouzon syndrome patients were screened for mutations in the two exons of FGFR2 by polymerase chain reaction and direct sequencing. RESULTS: Mutations were detected in 67% (6/9) of all cases. More than half the studied Crouzon patients carried a mutation resulting in either the loss or gain of a cysteine residue. A novel mutation, Tyr281Cys substitution, was discovered at exon IIIa. CONCLUSIONS: The mechanisms by which the same genotypes cause different phenotypes for each type of craniosynostosis syndrome in still uncertain. However, the molecular identification of the FGFR gene has made a great impact on the clinical classification of craniosynostosis syndromes; a new classification based on genotypes seems to be unavoidable. PMID- 11380922 TI - Uroguanylin level in umbilical cord blood. AB - BACKGROUND: Uroguanylin is a novel natriuretic and diuretic peptide originally isolated from urine. METHODS: To determine whether uroguanylin has a physiologic role during the perinatal period, uroguanylin levels in umbilical cord plasma obtained at the time of delivery were measured by radioimmunoassay and compared with cord serum osmolality. RESULTS: Mean (+/- SD) cord plasma uroguanylin concentrations (8.8 +/- 2.1 fmol/mL) were higher compared with normal adult values. The extent of maturity, mode of delivery and gender did not appear to influence cord uroguanylin levels. The uroguanylin concentration had a significant positive correlation with cord serum osmolality. CONCLUSION: These findings support some regulatory role of this peptide in perinatal renal and cardiovascular adaptation. PMID- 11380923 TI - Cyclobilirubin formation by in vitro photoirradiation with neonatal phototherapy light. AB - BACKGROUND: The main mechanism of phototherapy for neonatal hyperbilirubinemia is the production and excretion of (EZ)- and (EE)-cyclobilirubin (4E,15Z- and 4E,15E cyclobilirubin). Thus, the clinical efficacy of the light source for phototherapy must be evaluated by cyclobilirubin formation from (ZZ)-bilirubin in in vitro photoirradiation. METHODS: In the present study, we investigated the in vitro production pattern of bilirubin photoisomers by phototherapy light from the bilirubin-human serum albumin complex. RESULTS: No clear difference was found in the curves relative to (ZZ)-bilirubin and its photoisomers under aerobic and anaerobic conditions. The ratio of (EZ)-cyclobilirubin to (ZZ)-bilirubin increased proportionately to the dose of irradiating light and no photoequilibrium state was observed analogous to that found in configurational photoisomerization. The concentration of (EZ)- and (EE)-cyclobilirubin increased proportionately with the grade of the percentage decrease in A(460 nm) from 0 to 23%. With a percentage decrease in A(460 nm) of 23% or more, the cyclobilirubin concentrations reached a steady state. The reason for this appears to be that the concentration of (ZZ)-bilirubin, a substrate for photoisomers, dropped below 1 mg/100 mL. Biliverdin was produced only in trace amounts. However, the absorption at 520--700 nm increased after a percentage decrease in A(460 nm) of more than 23%. CONCLUSIONS: The results of the present study show that little bilirubin photooxidation occurred with in vitro aerobic photoirradiation. Before the concentration of cyclobilirubin reaches a steady state, it is theoretically valid to use the percentage decrease in A(460 nm) for the evaluation of the clinical efficacy of the light source. PMID- 11380924 TI - Evaluation of DNA damage using the comet assay in children on long-term benzathine penicillin for secondary prophylaxis of rheumatic fever. AB - BACKGROUND: Benzathine penicillin is the most widely used antibiotic in the prophylaxis of children with rheumatic fever. The aim of the present study was to evaluate the DNA damage in children receiving one dose of 1.2 million units benzathine penicillin every 4 weeks over a long period to prevent recurrences of rheumatic fever. METHODS: Thirty-five children with confirmed rheumatic fever under benzathine penicillin prophylaxis were enrolled in the study and 35 healthy children with similar ages and socioeconomic backgrounds served as controls. To detect any DNA damage, the comet assay was performed on circulating lymphocytes from the study subjects. RESULTS: Damaged (limited and extensive migration) cells in children on prophylactic therapy were higher than those in controls (P<0.001). CONCLUSIONS: It has been suggested that differences in the comet scores were induced by the administration of benzathine penicillin over a long period of time and further investigations are needed to confirm this toxic effect. PMID- 11380925 TI - A study of mismatch negativity in newborns. AB - BACKGROUND: Although newborns are presumed to have cognitive ability, studies on the objective cognitive ability of neonates are making little progress. METHODS: We gave auditory and visual stimuli to newborns whose conceptional age ranged from 35 to 43 weeks and recorded the mismatch negativity (MMN) that is said to reflect the process by which stimuli are detected automatically. We compared the waveform and latency of the MMN. RESULTS: The mismatch negativity waveform in newborns was similar to that of adults. The MMN latency tended to shorten with an advance in conceptional age and was especially shortened between 36 and 37 weeks of fetal life. In some newborns whose clinical course indicated a possibility of their having cognitive dysfunction, the MMN latency was prolonged. CONCLUSIONS: Newborns also have cognitive ability and MMN is useful for objectively evaluating the cognitive ability in newborns. PMID- 11380926 TI - Glomerulopathy with mesangial IgM deposits: long-term follow up of 64 children. AB - BACKGROUND: The aim of the present study was to investigate to what extent IgM nephropathy in children with minimal change nephrotic syndrome (MCNS) and diffuse mesangial hypercellularity (DMH) evolves to focal segmental glomerulosclerosis (FSGS). METHODS: Tissues from renal biopsies were examined by light microscopy (LM), immunofluorescence (IF) and, in four cases, by electron microscopy (EM). From a total of 352 nephrotic children, 121 had renal biopsy results as steroid dependent or resistant. A diagnostic renal biopsy was also performed in 331 children with non-nephrotic proteinuria and/or hematuria. A second renal biopsy was performed in 16 children whose renal function was impaired during the follow up. The clinical course of IgM-positive children was compared with that of IgM negative children. RESULTS: Of the 121 nephrotic children with renal biopsy, 85 were MCNS. Twenty were IF positive mainly for IgM, six of whom (30%) presented evolution to FSGS, while of the remaining 65 IF-negative children, only three (4.6%) presented evolution to FSGS. Of the total 331 children with non-nephrotic proteinuria and/or hematuria, 139 were diagnosed as IgA--IgG nephropathy, 44 had positive IF for IgM and 148 were IF negative. Of the 44 children IF positive for IgM, seven (15.9%) presented evolution to FSGS, while none of the 148 IF-negative children presented evolution to FSGS. The follow-up time for all children ranged from 1 to 14 years. CONCLUSIONS: Of IgM nephropathy patients with MCNS and DMH, a significant percentage develop impaired renal function, due to the evolution of FSGS, as revealed by repeat biopsy during long-term follow up. PMID- 11380927 TI - Trp290Cys mutation of the FGFR2 gene in a patient with severe Pfeiffer syndrome type 2. PMID- 11380928 TI - Schizencephaly in triple-X syndrome. PMID- 11380929 TI - Griscelli syndrome: report of a case and review of the literature. PMID- 11380930 TI - Hepatoblastoma associated with trisomy 18 syndrome: a case report and a review of the literature. PMID- 11380931 TI - Successful human leukocyte antigen one antigen-mismatched related bone marrow transplantation in a 6-year-old boy with leukocyte adhesion deficiency syndrome. PMID- 11380932 TI - Senior-Loken syndrome associated with mental retardation and microcephaly. PMID- 11380933 TI - Distal 14q trisomy due to a maternal derivative chromosome 14. PMID- 11380934 TI - Improvement in daily activities using a portable ventilator in a patient with spondyloepiphyseal dysplasia congenita. PMID- 11380935 TI - A rare case of congenital hepatic fibrosis with severe pulmonary hypertension in an adolescent. PMID- 11380936 TI - Incidence of Haemophilus influenzae type b meningitis in Mie prefecture, Japan. PMID- 11380937 TI - HLA-DPB1 typing by polymerase chain reaction amplification with sequence-specific primers. AB - DPB1 is the second most polymorphic class II locus with currently 84 recognized alleles, i.e. DPB1*0101 to DPB1*8101. Most of the alleles have been described during the last few years using oligonucleotide and sequencing techniques and relatively little is known about the role and importance of the polymorphic residues as regards to the function of DP molecules. In the present study, polymerase chain reaction (PCR) primers were designed for identification of all the phenotypically different DPB1 alleles by PCR amplification with sequence specific primers. Forty-eight standard genomic PCR reactions per sample were performed in order to achieve this resolution. Unique amplification patterns were obtained in 2983 of 3160 (94.4%) possible genotypes. The primers were combined so that only very rare genotypes gave rise to ambiguous patterns. Sixty-four Histocompatibility Workshop cell lines and 150 DNAs provided by the UCLA DNA exchange were investigated by the DPB1 primer set. All typing results were conclusive. Analysis of the distribution of DPB1 alleles was performed in 200 Caucasian samples, 100 African samples and 40 Oriental samples. The population study by the DPB1 PCR-SSP method showed a characteristic distribution of HLA-DPB1 alleles. Each ethnic group had one, or two, frequent DPB1 allele(s) and the frequency of homozygotes was high, suggesting that balancing selection does not appear to be affecting the evolution of the DPB1 locus. PMID- 11380938 TI - Resolution of cis-trans ambiguities between HLA-DRB1 alleles using single-strand conformation polymorphisms and sequencing. AB - DNA-based typing of HLA alleles occasionally results in the inability to assign a specific allele because of ambiguity in associating two or more polymorphisms to the same or to alternate homologs (cis/trans ambiguity). Since most individuals are heterozygous at a given HLA locus, the highest level of confidence in definition is obtained when the alleles are tested in isolation. By using single strand conformation polymorphisms (SSCP) to separate heterozygous HLA-DRB1 alleles, followed by sequencing of separated conformers, we have achieved resolution of previously ambiguous assignments without the need for additional probes or primers. PMID- 11380939 TI - HLA alleles and haplotypes in the Turkish population: relatedness to Kurds, Armenians and other Mediterraneans. AB - Turkish and Kurdish HLA profiles are studied for the first time. The comparative study of their allele frequencies, characteristic haplotypes, genetic distances with other Mediterraneans is complemented by neighbor-joining dendrograms and correspondence analyses. Turks, Kurds, Armenians, Iranians, Jews, Lebanese and other (Eastern and Western) Mediterranean groups seem to share a common ancestry: the older "Mediterranean" substratum. No sign of the postulated Indo-European (Aryan) invasion (1200 B.C.) is detected by our genetic analysis. It is concluded that this invasion, if occurred, had a relatively few invaders in comparison to the already settled populations, i.e. Anatolian Hittite and Hurrian groups (older than 2000 B.C.). These may have given rise to present-day Kurdish, Armenian and Turkish populations. PMID- 11380940 TI - Single nucleotide polymorphisms in intron 2 of the human interleukin-1 receptor antagonist (IL-1Ra) gene: further definition of the IL-1 beta and IL-1Ra polymorphisms in North American Caucasians and Taiwanese Chinese. AB - Previous studies have suggested that a variable number tandem repeat (VNTR) polymorphism in the second intron of the interleukin-1 receptor antagonist (IL 1Ra) gene and the single nucleotide polymorphisms at positions -511 and +3954 of the IL-1beta gene might be associated with increased risks of chronic inflammatory diseases, autoimmune diseases and gastric cancer. In the present study, IL-1beta and IL-1Ra genotypes were analyzed among Asians in Taiwan and Caucasians in North America. We identified a novel polymorphism with 3 nucleotide substitutions in the IL-1Ra VNTR 2-repeat allele. One of the substitutions corresponds with the fourth 3' end nucleotide of the reverse primer that is often used for analysis of the IL-1Ra-associated VNTR locus. Mismatching between this primer and the 2-repeat allele can cause misleading amplification results when stringent conditions are used for annealing. The estimated haplotype frequencies of the variant IL-1 genes were significantly different between Taiwanese and Caucasians. The frequency of the pro-inflammatory IL-1Ra 2-repeat allele was significantly lower in Taiwanese than in Caucasians. In contrast, the frequencies of the pro-inflammatory IL-1beta -511T allele and +3954C allele were significantly higher among Taiwanese compared with Caucasians. PMID- 11380941 TI - Tissue distribution of the human CD97 EGF-TM7 receptor. AB - CD97 is a founding member of the EGF-TM7 family of class II seven-span transmembrane (7-TM) receptors. CD97 has an extended extracellular region with several N-terminal epidermal growth factor (EGF)-like domains, which mediate binding to CD55. Previous studies demonstrated the expression of CD97 on activated lymphocytes, monocytes, macrophages, granulocytes, and numerous haematopoietic and nonhaematopoietic cell lines. Here, we determined the cellular distribution of human CD97 in situ by immunohistochemistry (IH) and immunofluorescence (IF). Abundant expression of CD97 was detected on all types of macrophages and dendritic cells, except for microglia. Within the lymphoid lineage, most T cells but only a few B cells express CD97. Germinal centre B cells do not express the molecule. Except for smooth muscle cells, no staining was found on other cells outside the immune system. However, analysis of a restricted set of epithelial tumors revealed CD97 expression on the malignant cells in thyroid and gastrointestinal tract cancer. PMID- 11380942 TI - Improved typing procedure for the polymorphic single-copy RLA-DQA gene of the rabbit reveals a new allele. AB - The DQA gene of the rabbit major histocompatibility complex (MHC, RLA) is highly polymorphic and, in contrast to those reported for other mammalian species, is present as a single copy. These properties allow use of this gene in a method to type the class II locus of RLA by a combination of single-stranded conformational polymorphism (SSCP) and heteroduplex (HD) analysis. Familial segregation of RLA DQA was shown and RLA class II types for rabbits of unknown pedigree were determined using migration patterns of amplified genomic DNA. Typing results were confirmed in experiments where unknown samples were mixed with products from rabbits of RLA types defined by sequence analysis. These analyses detected an RLA DQA allele in addition to the five previously described; this new allele is designated RLA-DQA-F. PMID- 11380943 TI - HLA-B allele frequencies in Cote d'Ivoire defined by direct DNA sequencing: identification of HLA-B*1405, B*4410, and B*5302. AB - Direct automated DNA sequencing was used to analyze exons 2 and 3 of HLA-B alleles present in forty-four unrelated individuals residing in the village of Adiopodoume, Cote d'Ivoire (Ivory Coast). Of the 23 HLA-B alleles observed, the most frequently detected allele was HLA-B*5301 (22.7%), which is believed to confer resistance to severe Plasmodium falciparum malaria. B*4501 (9.1%), B*1503 (8.0%), B*0705 (5.7%), B*1510 (5.7%) and B*3501 (5.7%) occurred frequently in the population. A second allele of B53 was identified; B*5302 contains a single amino acid variation at residue 171 (Y-->H). Two additional novel alleles, B* 1405 (a single amino acid variant of B*1402) and B*4410 (a five amino acid variant of B*4403) were characterized. PMID- 11380944 TI - Protective and susceptible HLA polymorphisms in IgA nephropathy patients with end stage renal failure. AB - Idiopathic immunoglobulin A (IgA) nephropathy is characterised by an extreme variability in clinical course, leading to end-stage renal failure in 15-20% of adults. This subgroup of patients with IgA nephropathy is usually included in the waiting lists of organ exchange organisations. The frequency of HLA-A,B,DR antigens of this subset of IgA nephropathy patients was calculated and compared to controls. The antigens HLA-B35 and DR5 were significantly increased in the patients with relative risk values of 1.385 and 1.487, respectively. The antigens HLA-B7, B8, DR2, and DR3 were found in a significantly lower frequency in the patients as compared to the controls. The relative risk (RR) values ranged between 0.695 and 0.727. Consequently, the haplotypes HLA-A1, B8, DR3, HLA-A3, B7, DR2, HLA-A2, B7, DR2 together with HLA-A1, B15, DR4, HLA-A9, B12, DR7, and HLA-A10, B18, DR2 were found to be protective with RR values ranging from 0.309 to 0.587. The only susceptible haplotype observed was HLA-A2-B5, DR5 (RR=2.990). PMID- 11380945 TI - HLA class II antigens in South African Blacks with type I diabetes. AB - Type 1 diabetes mellitus is poorly characterised in many African communities, including South Africa, where little is known of the disease epidemiology. This study aimed to identify the HLA class II alleles associated with type 1 diabetes in a group of Zulu subjects in Durban, KwaZulu-Natal by PCR-SSP. The HLA alleles associated with type 1 diabetes included HLA-DQB*0302 (P<0.0001), DRB1*O9 (P<0.0001), DRB1*04 (P=0.002), DRB1*0301 (P=0.003), DQB*02 (P=0.004) and DQA*03 (P=0.035). Estimated haplotypes positively associated with type 1 diabetes included HLA-DRB1 *0301-DQA*0501, DRB1*04-DQA*03, DRB1*04-DQB*0302, DRB1*0301 DQB*0201, DQA*0501-DQB*0201 and DQA*03-DQB*0302. These findings are similar to those reported from Zimbabwe and other populations with type 1 diabetes. PMID- 11380946 TI - Lack of association between NRAMP1 gene polymorphisms and Trypanosoma cruzi infection. AB - Genetic analysis in mice and humans have established the key role of the human natural resistance-associated macrophage protein 1 (NRAMP1) in resistance to intracellular infections. In the present study we investigated whether four NRAMP1 polymorphisms (5'(GT)n, -236 C-->T, D543N, and 3'UTR deletion) were important in determining the susceptibility to Trypanosoma cruzi infections as well as in the development of chagasic cardiac disease. Genotyping for these variants was assessed in 83 seropositive (asymptomatic, n=51, cardiomyopathic, n=32) and 85 seronegative individuals from a Peruvian population where T. cruzi is endemic. No statistically significant differences either between patients and controls or between asymptomatic and cardiomyopathic individuals were observed with respect to NRAMP1 variants. Our data suggest that the NRAMP1 genetic polymorphism analysed do not play a major role in the pathogenesis of T. cruzi infection in this Peruvian sample. PMID- 11380947 TI - Genomic diversity of natural killer cell receptor genes in three populations. AB - We report the distribution of genes encoding 11 killer cell immunoglobulin-like receptors (KIR) and 2 CD94:NKG2 receptors, in 32 Caucasians, 67 Australian Aborigines and 59 Vietnamese. The inhibitory and the activating KIR genes were found at different frequency in the three populations. No correlation was found between the polymorphism of the KIR genes and the HLA specificities of the tested samples. The most significant KIR associations were 2DL2 with 2DS2; 2DL2 with 2DS3 and 3DL1 with 2DS4 in all three study groups. In Caucasians and Vietnamese 2DS2 was associated with 2DS3 and 2DS1with 3DS1. KIR 2DL1 was strongly associated with three other KIRs: 2DL3, 3DL1 and 2DS4 in Aborigines. The distribution of the KIR phenotypes was different in the three populations. The AA1 phenotype was frequent in Vietnamese (42.4%) and Caucasians (31.2%), but very rare in Aborigines (1.5%). In contrast, the BB7 phenotype was very common for Aborigines (22.4%) and was absent in the two other groups. Our data demonstrate that different associations and putative KIR haplotypes could be distinguished in different populations. PMID- 11380949 TI - A novel HLA-A*02 allele, A*02202, identified by nucleotide sequencing. AB - In this paper we report the identification of a new HLA-A*02 allele in two members of an Afghan family. This novel allele, designed as A*02202, differs from A*02201 by a silent substitution at codon 66 (AAC-->AAT) in the alpha1 domain. A*02202 appears to be the result of a novel mutation (Note). PMID- 11380948 TI - Evidence for a novel polymorphism affecting both N-linked glycosylation and ligand binding of the IgG receptor IIIB (CD16). AB - Immunoglobulin G Fc receptor IIIb (FcgammaRIIIb) is constitutively expressed on neutrophils, and has three allelic forms: FcgammaRIIIb-NA1, FcgammaRIIIb-NA2, and FcgammaRIIIb-SH. We identified two Japanese subjects in whom an A to G substitution at nt 221 changes asparagine (N) to serine (S) at amino acid position 45 in the FcgammaRIIIb-NA2 gene. FcgammaRIIIb-NA2-specific monoclonal antibodies (GRM1 and PEN1) did not bind to mutant neutrophils, which lack an N linked glycosylation site. Furthermore, IgG3-mediated neutrophil phagocytosis by mutant was slightly increased as compared to wild-type donors (Note). PMID- 11380950 TI - Non-expression of HLA-B*5111N is caused by an insertion into the cytosine island at exon 4 creating a frameshift stop codon. AB - The identification of the "blank" allele HLA-B*5111N, which was detected in German and Czech individuals, is described. In the pedigree analysis this new allele segregates with the serological haplotype HLA-A2; B-; DR4 which is frequent in Czech population. The non-expression of B*5111N is caused by the insertion of an additional cytosine molecule at the cytosine island between the nucleotides 621-626 (codons 183-185, first three codons of exon 4) leading to a frame shift that creates a stop codon at codon 196. This insertion may be explained either by conversion with the pseudogene HLA-J or by slipped-strand mispairing. In order not to overlook the presence of alleles with altered expression in case of hematopoietic stem cell transplantation, both serological and DNA-based typing should be performed (Note). PMID- 11380951 TI - Novel HLA-B alleles associated with antigens in the 8C CREG. AB - This paper describes 13 novel HLA-B locus alleles, B*0809, B*0812, B*0813, B*0814, B*14062, B*3804, B*3806, B*3914, B*3915, B*3918, B*3919, B*3920, and B*3922 which represent new patterns of known polymorphic residues. PMID- 11380952 TI - New alleles in the B44 family including B*44022, B*44032, B*4411, B*4420, B*4421, B*4424, and B*8301. AB - Seven new HLA-B locus alleles have been described. B*44022 and B*44032 are silent substitutions altering known alleles. B*4411 carries a unique Bw4-like epitope. B*4420, B*4421, and B*4424 carry new combinations of motifs previously observed in other alleles. B*8301 appears to be the result of the replacement of exon 2 from B*4402 with exon 2 from B*5603. PMID- 11380953 TI - Novel HLA allele, HLA-B*4013, identified from a potential bone marrow recipient. Putative gene conversion events with donor sequence. AB - A new B40 allele was identified in a leukemic Caucasian patient. This allele, designated B*4013, differs in alpha 1 domain from B*4002 at six amino acidic positions: 67, 77, 80, 81, 82 and 83. Most of this substitutions could alter the antigen binding site of the HLA-B molecule. B*4013 may have originated by gene conversion or reciprocal recombination involving B*4002 as the recipient allele of sequence donated by B*4406. The new allele was serologically typed as a "blank" associated with the Bw4 epitope. PMID- 11380954 TI - Polymorphism of human HLA-DRB1 antigens generated by genetic exchange between DR2 (DRB1*15011) and DR6 (DRB1*1405) alleles: a novel DRB1 allele (DRB1*1437) identified in a Paiwan tribe member of Taiwan. AB - We report herein the identification of a new DRB1 allele using sequence-based typing (SBT). This novel allele, HLA-DRB1*1437, was found in an aboriginal individual from the Paiwan tribe in the southern part of Taiwan. This individual was typed by SBT method as having an HLA genotype of HLA-A*02011/0203, HLA B*15011/3901, HLA-DRB1*11011/1437, HLA-DRB3*0202/0202, and HLA-DPB1*0501/1301. This new allele differs from DRB1*1309 in the 5'-end nucleotide sequence of polymorphic exon 2 at codon 16 (CAT-->CAA; H16Q), codon 37 (AAC-->TTC; R37F), codon 47 (TTC-->TAC; F47Y), and codon 58 (GCC-->GCT; both specify alanine). By sequence comparison, it was found that this new allele has a 5'-end sequence (from amino acid residues 7 to 66) identical to that found in the DRB1*1405 allele and a 3'-end sequence (from amino acid residues 58 to 94) identical to that found in the DRB1*15011 allele. Both DRB1*1405 and DRB1*15011 alleles have been identified among the Paiwan members (Note). PMID- 11380955 TI - Description of a new HLA-DRB1 allele, DRB1*1139. AB - We report the identification of a novel DRB1*11 using sequence-based typing. This new allele, officially named DRB1*1139, was detected while performing HLA-DRB1 high-resolution typing of a volunteer bone marrow donor. DRB1*1139 is identical to DRB1*11011 except at codon 51 (ACG-->AGG) changing the encoded Threonine to Arginine. The triplet AGG has never been found in any other DRB1 allele. In fact, with standard polymerase chain reaction (PCR) amplification with sequence specific primers, the presented allele would have been interpreted as DRB1*1101 (Note). PMID- 11380956 TI - Identification of three new DRB3* (DRB3*0106, DRB3*0107 and DRB3*02022) alleles. AB - Three novel DRB3* alleles were identified using CANTYPE reverse hybridization assay. The initial unusual hybridization patterns of DRB3-specific polymerase chain reaction (PCR)-amplified DNA from each subject were confirmed by cloning and sequencing analysis. DRB3*0106 allele is identical to DRB3*0101 except for a single nucleotide substitution (CTG-->GTG) changing codon 38 from Leu to Val. This polymorphism is commonly found in DRB3*03 alleles. Compared with DRB3*0202, DRB3*02022 contains a single silent nucleotide substitution (AAT-->AAC, both encoding for Asn) at codon 77. This polymorphism is also present in DRB3*0204 allele. The new DRB3*0107 allele has a sequence unique to DRB3 alleles. From codon 5 to codon 36 the sequence is identical to that of DRB3*0101 allele. From codon 37 to codon 87 the sequence of DRB1*0107 allele is identical to that of DRB3*0202. This sequence would thus explain the CANTYPE(R) DRB3-specific unusual pattern of reactions. The new DRB3*0107 could have arisen from a gene conversion between DRB3*0101 and DRB3*0202 alleles, but the DRB3*0106 and the DRB3*02022 may have been generated by a point mutation event. The DRB3*0107 allele was identified in a Caucasoid individual. The ethnic origin of the subjects carrying the other two alleles are unknown. The three alleles presented here were only identified once, in a total population of 49,000. PMID- 11380957 TI - Nomenclature for factors of the HLA system, update January 2001. PMID- 11380958 TI - Examination of the fetal heart by five short-axis views: a proposed screening method for comprehensive cardiac evaluation. PMID- 11380959 TI - Isolated major congenital heart disease. PMID- 11380960 TI - Comparative analysis of pattern, management and outcome of pre- versus postnatally diagnosed major congenital heart disease: a population-based study. AB - OBJECTIVES: Most pregnant women in New South Wales undergo obstetric ultrasound examination, including some assessment of fetal cardiac anatomy. We aimed to review the spectrum of cardiac defects, management and outcome data of all fetuses with diagnosis of major congenital heart disease between 1994 and 1996 and compare them to major congenital heart disease in infants born during the same 3-year study period. METHODS: Descriptive comprehensive study of the New South Wales population. Study centers included the single fetal echocardiographic referral service and the two pediatric cardiac centers of New South Wales. RESULTS: Ninety-seven fetuses and 562 infants with major congenital heart disease were identified (240,000 livebirths), resulting in a prenatal detection rate of 15%. Anomalies detectable by cardiac four-chamber views were diagnosed at an average rate of 30% (68/229) in utero. By contrast, lesions associated with abnormal ventricular outflow and great artery views were detected in only 6.7% (29/430; P < 0.0001) of cases prior to birth. Of the 97 fetuses, 29 were aborted, 16 died in utero, and 9 died early postnatally without treatment. Within 2 weeks of age, 23% with fetal and 40% (P < 0.05) with infant major congenital heart disease diagnosis required an intervention, mainly for patent ductus arteriosus dependent lesions. Postnatal survival was similar for the fetal and infant series up to 2 years of age: 77% (95% confidence interval 64-90%) vs. 85% (95% confidence interval 82-88%). CONCLUSIONS: Prenatal diagnosis has important implications for pregnancy outcome, in particular for univentricular lesions. However, the present mode of obstetric routine ultrasound scanning fails to identify most ductus arteriosus dependent cardiac lesions with a predictable need for early postnatal intervention. PMID- 11380961 TI - Evaluation of prenatal diagnosis of congenital heart diseases by ultrasound: experience from 20 European registries. AB - OBJECTIVES: To evaluate prenatal diagnosis of congenital heart diseases by ultrasound investigation in well-defined European populations. DESIGN: Data from 20 registries of congenital malformations in 12 European countries were included. The prenatal ultrasound screening programs in the countries ranged from no routine screening to three ultrasound investigations per patient routinely performed. RESULTS: There were 2454 cases with congenital heart disease with an overall prenatal detection rate of 25%. Termination of pregnancy was performed in 293 cases (12%). There was considerable variation in prenatal detection rate between regions, with the lowest detection rates being in countries without ultrasound screening (11%) and in Eastern European countries (Croatia, Lithuania and Ukraine; 8%). In Western European countries with ultrasound screening, detection rate ranged from 19-48%. There was a significant difference in prenatal detection rate and proportion of induced abortions between isolated congenital heart disease and congenital heart disease associated with chromosome anomalies, multiple malformations and syndromes (P < 0.0001). There were 1694 cases with isolated congenital heart disease of which 16% were diagnosed prenatally. Malformations affecting the size of the ventricles were detected prenatally in half of the cases. CONCLUSIONS: Prenatal detection rate of congenital heart disease varies significantly between countries even with the same screening recommendations. The presence of associated malformations significantly increases the prenatal detection rate. PMID- 11380962 TI - Correlation between fetal cardiac diagnosis by obstetric and pediatric cardiologist sonographers and comparison with postnatal findings. AB - OBJECTIVE: To assess the level of agreement between obstetric and pediatric cardiologist sonographers' diagnosis of fetuses with suspected congenital heart disease and to compare this with the final postnatal diagnosis. DESIGN: We retrospectively reviewed the notes of 1037 patients undergoing fetal echocardiography over a 5-year period (1995-99) at the principal tertiary referral center for fetal cardiology in New South Wales, Australia. The median gestational age at presentation was 21 weeks (range 17-38) with 49% of the scans performed at < 21 weeks and a further 17% performed at 21-24 weeks. The accuracy of the fetal cardiac diagnosis was validated by neonatal cardiac investigation or autopsy. RESULTS: A total of 249 cases of congenital heart disease were identified antenatally during this 5-year period with the majority (84%) referred because of suspicion of a cardiac malformation during an obstetric scan. Of the 268 fetuses with congenital heart disease suspected by obstetric sonographers, 209 had confirmed cardiac defects. Complete correlation between obstetric sonographers' and pediatric cardiologists' prenatal cardiac findings was achieved in 62% of cases. There were major differences involving the atrioventricular morphology in 18% of cases and the outflow tract anatomy in 20%. Complete agreement between prenatal and postnatal diagnosis in fetuses with complex congenital heart disease was achieved in 59% of cases for obstetric sonography (17% false positive; 41% false negative) and 95% for fetal echocardiography by pediatric cardiologists (2% false positive; 5% false negative). CONCLUSIONS: Improved accuracy in diagnosis can be achieved through a pediatric cardiologist with special skills in fetal echocardiography working collaboratively with obstetric sonographers to optimize the details of diagnosis. This may influence management and counseling. PMID- 11380963 TI - Arterial Doppler ultrasound in 115 second- and third-trimester fetuses with congenital heart disease. AB - OBJECTIVE: To assess the influence of isolated congenital heart disease (CHD) on fetal arterial Doppler blood flow velocity waveforms. METHODS: Doppler flow velocimetry was performed in the umbilical artery and middle cerebral artery in 115 consecutive fetuses with antenatally diagnosed CHD. Gestational age ranged between 19 and 41 weeks. Fetuses with isolated CHD were defined as group A (n = 55), showing cardiogenic hydrops fetalis in six cases; group B included 60 cases complicated by chromosomal or non-chromosomal extracardiac malformation, uteroplacental dysfunction or non-cardiogenic non-immune hydrops fetalis. The control group comprised 100 healthy fetuses of uncomplicated pregnancies. Individual pulsatility index measurements were converted into their Z-scores (delta values) for statistical analysis. RESULTS: In regard to the umbilical artery pulsatility index, 115 fetuses with CHD showed a significantly greater (P < 0.001) difference from the normal mean for gestation (delta values) than the control group. However, 29 of the 33 cases with indices above the 95% reference interval were additionally associated with extracardiac malformations, uteroplacental dysfunction or non-cardiogenic non-immune hydrops fetalis. While fetuses with isolated CHD still showed significantly higher values than healthy fetuses (P < 0.01), only in 4 of 55 (7%) fetuses did the measured umbilical artery pulsatility index exceed the 95% reference interval. There was no significant difference from the control group, in which 4 of 100 cases showed an umbilical artery pulsatility index above the 95% reference interval. Elevated umbilical artery pulsatility indices were seen in only four cases of severe obstruction of the outflow tracts leading to reverse perfusion of the affected great artery and in one case of Ebstein's anomaly with pulmonary insufficiency. Although all four fetuses with isolated CHD and elevated umbilical artery pulsatility index died, 14 of 18 fetuses with lethal outcome had normal pulsatility index values in the umbilical artery. Investigations of the middle cerebral artery blood flow revealed no significant difference between fetuses with and without CHD or any subgroups. CONCLUSIONS: This study shows that arterial blood flow velocity waveforms in fetuses with isolated CHD do not show sufficient alterations to be of diagnostic value. Only in severe outflow tract obstructions due to a 'steal effect' or in significant insufficiencies of semilunar valves leading to an impaired 'wind-kessel function' may the special hemodynamic changes induced by CHD result in a significant increase of pulsatility index in the umbilical artery. In the majority of cases with CHD the increase of pulsatility index of umbilical arterial blood flow velocity waveforms, however, results from extracardiac anomalies, especially uteroplacental dysfunction and chromosomal abnormalities. Furthermore, umbilical artery Doppler sonography is not clinically helpful in predicting fetal outcome. PMID- 11380964 TI - Fetal hydrops and hepatosplenomegaly in the second half of pregnancy: a sign of myeloproliferative disorder in fetuses with trisomy 21. AB - OBJECTIVE: To demonstrate the relationship between fetal hydrops and/or hepatosplenomegaly in the second half of pregnancy with a myeloproliferative disorder in fetuses with trisomy 21 or mosaic trisomy 21. DESIGN: A retrospective case series. SUBJECTS: Cases were selected from 79 cases of trisomy 21 diagnosed in our prenatal unit between 1993 and 1999. METHODS: All fetuses had a detailed sonographic anatomic survey and biometry. Doppler of the umbilical and middle cerebral arteries, ductus venosus, inferior vena cava and umbilical vein was performed whenever possible. Two-dimensional echocardiography supplemented by color Doppler flow mapping and spectral pulsed wave Doppler was performed in all cases of fetal hydrops. Fetal karyotyping was obtained by amniocentesis, chorionic villus sampling or fetal blood sampling. In the presence of fetal hydrops a cordocentesis was performed for fetal hematology, biochemistry and TORCH serology. In cases with diagnosis of myeloproliferative disorder, peripheral blast cells were characterized by microscopy, cytochemistry and determination of surface markers. All cases with myeloproliferative disorder were stillborn and subsequently had a postmortem examination performed. RESULTS: During the study period 79 cases of trisomy 21 were diagnosed. Eleven of these had fetal hydrops. Three of these fetuses presented with hepatosplenomegaly and myeloproliferative disorder in the second and third trimesters. In addition, one fetus with sonographic markers of trisomy 21, where karyotyping was unfortunately unsuccessful, presented with hepatosplenomegaly, hydrops and myeloproliferative disorder. In the four fetuses with hepatosplenomegaly and hydrops, serology was negative for congenital infection. The characteristics of blast cells in the peripheral blood smear revealed a myeloproliferative disorder. CONCLUSION: Fetal hydrops and/or hepatosplenomegaly in the second half of pregnancy, although suggestive of infectious etiology, may be a sign of myeloproliferative disorder in fetuses with trisomy 21 or mosaic trisomy 21. There is a possibility that a transient myeloproliferative disorder is a more common cause of mid or late trimester hydrops in cases of trisomy 21 than previously thought. In these hydropic fetuses the prognosis seems to be poor. On the other hand we can speculate that a myeloproliferative disorder and the associated hepatosplenomegaly and/or hydrops may show spontaneous remission or that the transient myeloproliferative disorder may be without any detectable ultrasonographic signs and therefore may be more frequent in utero than realized. PMID- 11380965 TI - Sonographic assessment of amniotic fluid volume between 11 and 24 weeks of gestation: construction of reference intervals related to gestational age. AB - OBJECTIVE: At present, most of the methods for sonographic assessment of amniotic fluid volume are unreliable in the second trimester of pregnancy, or else they do not present nomograms related to gestational age. DESIGN: The aim of this prospective cross-sectional study was to construct normal reference ranges of four ultrasound parameters for the evaluation of amniotic fluid volume which could be applied in the second trimester. For these parameters we calculated normal curve limits suitable for use in clinical practice. SUBJECTS: From a population of normal pregnant women between the 12th and the 24th weeks of gestation undergoing a routine ultrasound examination during 1997 at our institute, 273 were found to be suitable for the study, after the exclusion of all cases which presented any feto-maternal pathology or complications up to the 24th week. METHODS: The largest 'amniotic pocket' in a vertical direction, free of small fetal parts and umbilical cord, was measured: the maximum vertical and transverse diameters were measured on the same scan; the mean diameter and the product of the two diameters were calculated. The 'mean amniotic fluid diameter', the 'two-diameter pocket', the 'largest vertical pocket' and the 'largest transverse pocket' were the four sonographic parameters considered. RESULTS: The four parameters correlated well with gestational week and with the biparietal diameter; the normal reference intervals and normal curve were then calculated. All these parameters were found to have good intra- and interoperative reproducibility. CONCLUSIONS: We conclude that the use of an ultrasound semiquantitative method based on the measurement of a single amniotic fluid pocket and involving normal reference intervals according to gestational age could improve the early diagnosis of amniotic fluid variations during the second trimester, although this has yet to be confirmed by extensive clinical trials. PMID- 11380966 TI - Middle cerebral artery Doppler in severe intrauterine growth restriction. AB - OBJECTIVE: To examine longitudinal changes in middle cerebral artery blood flow assessed by Doppler in severely growth restricted fetuses. METHODS: Eighteen structurally normal singleton pregnancies complicated by suspected intrauterine growth restriction were monitored by serial measurement of the pulsatility index of the middle cerebral artery over 7 to 72 days. Outcome measures included indication for delivery, umbilical venous pH and admission to and length of stay in neonatal intensive care. RESULTS: Thirteen fetuses demonstrated severe intrauterine growth restriction based on subsequent birth weights being below the 2.5th centile, two had intrauterine growth restriction (birth weights between the 2.5th and 5th centiles), and three had birth weights between the 5th and 50th centiles. The middle cerebral artery pulsatility index showed rapid and sharp changes between examinations in those severely growth restricted fetuses which required delivery before 34 weeks. This pattern was not obvious in severely growth restricted fetuses delivered after 34 weeks, or in those less severely growth restricted, regardless of the gestation at delivery. Changes in middle cerebral artery pulsatility index contributed to the decision to deliver in three cases. The middle cerebral artery pulsatility index demonstrated greater variation in those fetuses with cord pHs of less than 7.25. The length of stay in neonatal intensive care decreased with increasing gestational age and birth weight. CONCLUSIONS: The difference in the pattern of change in middle cerebral artery pulsatility index in intrauterine growth restricted fetuses may be a reflection of maturity in addition to the degree of fetal compromise. The decision to deliver was multifactorial. The middle cerebral artery pulsatility index only influenced the decision to deliver when changes in other parameters were evident. PMID- 11380967 TI - Three-dimensional ultrasound: a novel technique for investigating the urethral sphincter in the third trimester of pregnancy. AB - OBJECTIVES: To measure urethral sphincter volume by three-dimensional (3D) ultrasound. To assess the reproducibility of this measurement technique and to compare volumes obtained using 3D ultrasound with volumes calculated from a formula based on 2D ultrasound measurements. METHODS: Women were recruited as part of an ongoing study of changes to the pelvis resulting from pregnancy and childbirth. One hundred and eleven women in the third trimester of pregnancy (between 32 and 41 completed weeks' gestation) underwent a 3D transvaginal ultrasound scan of the urethra. In 10 cases the scan was analyzed twice by different observers to assess the reproducibility of the measurements from the scans and the results were analyzed using limits of agreement. RESULTS: The interobserver error was consistent between all the linear, 2D and 3D measurements obtained from the area scanned. There was a significant difference between volumes calculated directly by 3D ultrasound and the approximated volumes from conventional 2D measurements. CONCLUSIONS: Three-dimensional ultrasound appears to be a useful tool in measuring urethral sphincter volume. The error is consistent with that of linear and 2D imaging. However, the increase in normal range generated by biological variation in all three planes makes 3D ultrasound a more sensitive method of evaluating change to the urethral sphincter. PMID- 11380968 TI - Three-dimensional ultrasound of the female urethra: comparing transvaginal and transrectal scanning. AB - OBJECTIVES: To compare transvaginal and transrectal three-dimensional ultrasound in determining the morphology and measurements of the female urethra. DESIGN: Sixty-five women who had not had surgery for incontinence or pelvic floor descent had transvaginal and transrectal sonography using a 7.5-MHz mechanical sector endoprobe with three-dimensional facilities. The multiplanar display of the scanned volumes allowed detailed morphologic assessment of the urethra and the measurement of distances and volumes. Statistical endpoints were: sagittal urethral diameter, maximum rhabdosphincter length and thickness, maximum thickness of the smooth muscle complex, and the volumes of the rhabdosphincter and the smooth muscle complex. Values were compared between the two approaches using Student's t-test and Bland-Altman analysis. RESULTS: Both vaginal and rectal scans were feasible. However, significant differences between the two approaches were found for the sagittal diameter of the urethra (8.4 +/- 1.9 mm on vaginal vs. 11.5 +/- 2.2 mm on rectal scans, P < 0.01) and the transverse diameter of the urethra's smooth muscle complex (11.2 +/- 0.3 mm on vaginal vs. 8.6 +/- 0.2 mm on rectal scans, P < 0.001). No other variables showed significant differences. Compression of the urethra and displacement under the symphysis pubis were observed when the ultrasound probe was applied vaginally. Bland-Altman analysis showed acceptable variability for differences of distances but considerable variability for the differences of volumes. CONCLUSION: The female urethra can be examined both vaginally and rectally by three-dimensional ultrasound. A transvaginally applied probe seems to have a compression effect on the urethra. PMID- 11380969 TI - Intraobserver reproducibility of Doppler measurements of uterine artery blood flow velocity in premenopausal women. AB - OBJECTIVE: To determine the intraobserver repeatability of Doppler measurements of uterine artery blood flow velocity and the contribution of various factors to within-subject variance. DESIGN: Seventeen healthy premenopausal women underwent vaginal Doppler ultrasound examination of the uterine artery by the same observer. Three measurements were taken at each of three sites: 1) the currently recommended sampling site; 2) the ascending branch of the uterine artery at a level between the lower and middle third of the corpus uteri; 3) 1.5 cm lateral to the recommended sampling site. Three measurements were taken at each site. For each measurement, three uniform consecutive cardiac cycles were analyzed. Peak systolic velocity, time-averaged maximum velocity, and pulsatility index were calculated. Each Doppler shift spectrum was analyzed twice. Thus, for each women, 18 measurement results per sampling site were obtained. Analysis of variance was used. RESULTS: The effect of sampling site on measurements of peak systolic velocity and time-averaged maximum velocity was non-significant, but pulsatility index values obtained at the distal sampling site were slightly higher than those obtained at the other sites (P = 0.01). Repetition accounted for most of the within-subject variance. Averaging the results of the three repeat measurements yielded increased intraclass correlation coefficients: 0.79-0.89 for peak systolic velocity, 0.80-0.92 for time-averaged maximum velocity and 0.86-0.93 for pulsatility index. CONCLUSION: As the effect of repetition on the results of Doppler measurements of uterine artery blood flow velocity is large, the average of several repeat measurements should be used to enhance measurement reproducibility. However, it is not worth doing more than one analysis of a Doppler shift spectrum, and it is not worth analyzing more than one cardiac cycle per spectrum. PMID- 11380970 TI - Transvaginal color Doppler assessment of venous flow in adnexal masses. AB - OBJECTIVE: To analyze the usefulness of transvaginal color Doppler assessment of venous flow in the differential diagnosis of adnexal masses. MATERIAL AND METHODS: Ninety-one consecutive patients (mean age: 46.6 years, range: 16-81 years) diagnosed as having an adnexal mass were evaluated by transvaginal color Doppler sonography prior to surgery. Color Doppler was used to detect and analyze the flow velocity waveform from arterial and venous blood flow within the tumor. For arterial signals the resistance index and peak systolic velocity, and for veins the maximum venous flow velocity, were calculated. Receiver operator characteristic curves were plotted to determine the best venous flow velocity cut off. According to our previous study using arterial Doppler, a tumor was considered as malignant when flow was detected and the lowest resistance index was < or = 0.45. Using venous Doppler a mass was considered as malignant when flow was detected and the venous flow velocity was > or = the best cut-off found on the receiver operator characteristic curve. Definitive histopathological diagnosis was obtained in all cases. Sensitivity, specificity, positive predictive value and negative predictive value for B-mode morphology (evaluation performed according to Sassone's scoring system), arterial Doppler, venous Doppler, and a combination of both arterial and venous Doppler were calculated. RESULTS: Twenty-five masses (27.5%) were malignant and 66 (72.5%) benign. Arterial and venous flow was found more frequently in malignant than in benign masses (92% vs. 41% (P < 0.001) and 72% vs. 21% (P < 0.001), respectively). The resistance index was significantly lower in malignant tumors (0.42 vs. 0.60, P = 0.0003). No differences were found in peak systolic velocity. Venous flow velocity was significantly higher in malignant masses (18.1 cm/s vs. 8.9 cm/s, P = 0.0006). The best cut-off of venous flow velocity was 10 cm/s. Sensitivity, specificity, positive predictive value and negative predictive value for morphology, arterial Doppler, venous Doppler, and the combination of both arterial and venous Doppler were 92%, 71%, 45%, 96%; 76%, 95%, 87%, 91%; 68%, 94%, 81%, 89%; and 88%, 91%, 79%, 95%, respectively. CONCLUSIONS: Our results indicate that preoperative evaluation by venous flow assessment of adnexal masses may be useful to discriminate between malignant and benign tumors. PMID- 11380971 TI - Prenatal diagnosis and follow up of congenital splenic cyst: a case report. AB - We report a case of congenital splenic cyst that was identified by prenatal sonography at 25 gestational weeks. Serial scans demonstrated an increase in size during the prenatal period but after birth the cyst progressively decreased in size until its complete regression at 6 months of age. PMID- 11380972 TI - Hypoechoic hepatomegaly associated with transient abnormal myelopoiesis provides clues to trisomy 21 in the third-trimester fetus. AB - We present two cases of transient abnormal myelopoiesis associated with trisomy 21 that had hypoechoic hepatomegaly during the late fetal period. Fetal chromosomal abnormality and fetal myeloproliferative disorder should be suspected in such cases. PMID- 11380973 TI - Prenatal diagnosis of fetal abdominal lymphangioma by ultrasonography. AB - We present a case of abdominal lymphangioma in a fetus together with a review of the literature. Diagnosis was made at 20 weeks' gestation by antenatal ultrasonography. In keeping with other reports, the lesion was located on the left and serial ultrasonography demonstrated rapid growth with extension into the lower extremity. PMID- 11380974 TI - Retrospective prenatal diagnosis of scimitar syndrome aided by three-dimensional power Doppler imaging. AB - Scimitar syndrome is a rare malformation of the arterial supply and venous drainage of the lung. We report the case of a fetus that presented with cardiac asymmetry and malposition of the fetal heart. Postnatally, scimitar syndrome was confirmed at cardiac catheterization. Retrospective reconstruction of three dimensional power Doppler volumes, obtained during fetal life, allowed direct visualization of the abnormal aortopulmonary collateral vessel. This had not been seen on conventional scans. This case demonstrates many of the strengths of three dimensional sonographic techniques for the delineation of complex vascular anatomy. It confirms that a prospective diagnosis of scimitar syndrome should be possible during fetal life. PMID- 11380975 TI - Ultrasound diagnosis and Doppler monitoring of a pelvic spleen in pregnancy. AB - We describe a patient with a pelvic spleen diagnosed during pregnancy and monitored through gestation which we believe to be the first reported case. A 40 year-old woman was referred at 8 weeks of gestation because of a chronic intense pain in the left iliac cavity which had spread to her lower back. Clinical examination revealed a poorly defined pelvic mass. Pelvic ultrasound demonstrated a gestational sac containing a viable embryo whose size was consistent with the period of amenorrhea. While the splenic area in the left hypocondrium was found to be empty, a homogeneous and elongate mass measuring 152 x 123 mm with a maximum thickness of 53.4 mm was observed in the left iliac cavity above the uterus. This mass, the ectopic spleen, was monitored by Doppler velocimetry at monthly intervals until delivery and no variation throughout gestation was observed; therefore, despite the occasional occurrence of heavy pain, it was possible to exclude circulatory complications such as thrombosis or torsion. Doppler ultrasound proved to be a useful tool for the differential diagnosis of this rare anatomical variation. PMID- 11380976 TI - Three-dimensional power Doppler ultrasound of the fetal great vessels. PMID- 11380986 TI - Evaluating genome dynamics: the constraints on rearrangements within bacterial genomes. AB - Inversions and translocations distinguish the genomes of closely related bacterial species, but most of these rearrangements preserve the relationship between the rearranged fragments and the axis of chromosome replication. Within species, such rearrangements are found less frequently, except in the case of clinical isolates of human pathogens, where rearrangements are very frequent. PMID- 11380987 TI - The nitrilase superfamily: classification, structure and function. AB - The nitrilase superfamily consists of thiol enzymes involved in natural product biosynthesis and post-translational modification in plants, animals, fungi and certain prokaryotes. On the basis of sequence similarity and the presence of additional domains, the superfamily can be classified into 13 branches, nine of which have known or deduced specificity for specific nitrile- or amide-hydrolysis or amide-condensation reactions. Genetic and biochemical analysis of the family members and their associated domains assists in predicting the localization, specificity and cell biology of hundreds of uncharacterized protein sequences. PMID- 11380990 TI - Should we abandon the concept of giving patients "positive" and "negative" screening results? Yes: this dichotomy can be misleading. PMID- 11380991 TI - Should we abandon the concept of giving patients "positive" and "negative" screening results? No: we should do better at explaining results. PMID- 11380992 TI - Caring for immigrant women. Physicians' roles range from detective work to advocacy. PMID- 11380993 TI - Why health insurers should pay for addiction treatment. Treatment works and would lead to net societal benefits. PMID- 11380996 TI - FDA targets snack foods industry over allergens. PMID- 11380997 TI - Pilot program tests distributing emergency contraception without a prescription. PMID- 11380998 TI - You can always pop a pill. PMID- 11380999 TI - "Tortured tube" sign. PMID- 11381000 TI - Delayed birth equals more cancers and preterm births. PMID- 11381002 TI - More evidence to reassure physicians and parents about vaccination. PMID- 11381003 TI - Do drug advertisements in Russian medical journals provide essential information for safe prescribing? AB - OBJECTIVE: To examine pharmaceutical advertisements in medical journals for their adequacy of information. METHODS: We selected a convenience sample of 5 major Russian medical journals covering different fields of medicine and different types of publications. We evaluated all the ads in all the issues of the selected journals published during 1998. We counted the number of appearances of trade, chemical, and generic names; indication and contraindication; pharmacologic group; safety warnings; and references. Counts in all categories were aggregated for each advertiser. RESULTS: There were 397 placements of 207 distinct advertisements. Only 154 placements (40%) mentioned the generic name, 177 (45%) mentioned any indication, 42 (11%) mentioned safety warnings and contraindications, 21 (5%) warned about drug interactions, and 8 (2%) provided references. The 6 companies responsible for the most ads on average provided less information than the other companies. CONCLUSIONS: Almost none of the drug ads published in Russian medical journals provide the basic information required for appropriate prescribing. This is despite the fact that in Russia, ads that omit essential information and that could lead consumers to misunderstandings about an advertised product are illegal. The arrival of drug advertising in Russia has brought little information and has been potentially damaging. PMID- 11381005 TI - "Educational" advertisements--I haven't seen one yet! PMID- 11381006 TI - Are guns used more by US civilians for self-defense or for intimidation? PMID- 11381007 TI - Genetic consultation. PMID- 11381008 TI - Menopause and hormone replacement: Part 1. Evaluation and treatment. PMID- 11381009 TI - Assessing diagnostic and screening tests: Part 1. Concepts. PMID- 11381010 TI - The search for a job: bringing focus to a daunting task. PMID- 11381011 TI - Treating children with asthma. A review of drug therapies. PMID- 11381013 TI - A simple mnemonic for the diagnostic criteria for post-traumatic stress disorder. PMID- 11381016 TI - Myth: codeine is a powerful and effective analgesic. PMID- 11381017 TI - How does the United States compare with the rest of the world in human sexual behavior? PMID- 11381019 TI - Culture and clothes. PMID- 11381018 TI - Immigrant women's health: initial clinical assessment. PMID- 11381020 TI - From comparative and functional genomics to practical decisions in the clinic: a view from the trenches. PMID- 11381021 TI - Application of SNP technologies in medicine: lessons learned and future challenges. PMID- 11381022 TI - Indian caste origins: genomic insights and future outlook. PMID- 11381024 TI - Evolutionary role of restriction/modification systems as revealed by comparative genome analysis. AB - Type II restriction modification systems (RMSs) have been regarded either as defense tools or as molecular parasites of bacteria. We extensively analyzed their evolutionary role from the study of their impact in the complete genomes of 26 bacteria and 35 phages in terms of palindrome avoidance. This analysis reveals that palindrome avoidance is not universally spread among bacterial species and that it does not correlate with taxonomic proximity. Palindrome avoidance is also not universal among bacteriophage, even when their hosts code for RMSs, and depends strongly on the genetic material of the phage. Interestingly, palindrome avoidance is intimately correlated with the infective behavior of the phage. We observe that the degree of palindrome and restriction site avoidance is significantly and consistently less important in phages than in their bacterial hosts. This result brings to the fore a larger selective load for palindrome and restriction site avoidance on the bacterial hosts than on their infecting phages. It is then consistent with a view where type II RMSs are considered as parasites possibly at the verge of mutualism. As a consequence, RMSs constitute a nontrivial third player in the host-parasite relationship between bacteria and phages. PMID- 11381023 TI - TTY2: a multicopy Y-linked gene family. AB - Genes involved in human male sex determination and spermatogenesis are likely to be located on the Y chromosome. In an effort to identify Y-linked, testis expressed genes, a cDNA selection library was generated by selecting testis cDNA with Y-cosmid clones. Resultant clones containing repetitive or vector material were eliminated, and 79 of the remaining clones were sequenced. Nineteen cDNAs showed homology with the TTY2 gene, and indicated that TTY2 is part of a large gene family. Screening of a panel of Y-linked cosmids revealed that the TTY2 gene family includes at least 26 members organized in 14 subfamilies. Further investigation revealed that TTY2 genes are arranged in tandemly arrayed clusters on both arms of the Y chromosome, and each gene comprises a series of tandemly arranged repeats. RT-PCR studies for two of these genes revealed that they are expressed in adult and fetal testis, as well as in the adult kidney. None of the genes investigated in detail contain an open reading frame. We conclude that the TTY2 gene family is composed of multiple copies, some of which may function as noncoding RNA transcripts and some may be pseudogenes. PMID- 11381025 TI - Genome-wide epistatic interaction analysis reveals complex genetic determinants of circadian behavior in mice. AB - Genetic heterogeneity underlies many phenotypic variations observed in circadian rhythmicity. Continuous distributions in measures of circadian behavior observed among multiple inbred strains of mice suggest that the inherent contributions to variability are polygenic in nature. To identify genetic loci that underlie this complex behavior, we have carried out a genome-wide complex trait analysis in 196 (C57BL/6J X BALB/cJ)F(2) hybrid mice. We have characterized variation in this panel of F(2) mice among five circadian phenotypes: free-running circadian period, phase angle of entrainment, amplitude of the circadian rhythm, circadian activity level, and dissociation of rhythmicity. Our genetic analyses of these phenotypes have led to the identification of 14 loci having significant effects on this behavior, including significant main effect loci that contribute to three of these phenotypic measures: period, phase, and amplitude. We describe an additional locus detection method, genome-wide genetic interaction analysis, developed to identify locus pairs that may interact epistatically to significantly affect phenotype. Using this analysis, we identified two additional pairs of loci that have significant effects on dissociation and activity level; we also detected interaction effects in loci contributing to differences of period, phase, and amplitude. Although single gene mutations can affect circadian rhythms, the analysis of interstrain variants demonstrates that significant genetic complexity underlies this behavior. Importantly, most of the loci that we have detected by these methods map to locations that differ from the nine known clock genes, indicating the presence of additional clock-relevant genes in the mammalian circadian system. These data demonstrate the analytical value of both genome-wide complex trait and epistatic interaction analyses in further understanding complex phenotypes, and point to promising approaches for genetic analysis of such phenotypes in other mammals, including humans. PMID- 11381026 TI - Genome evolution at the genus level: comparison of three complete genomes of hyperthermophilic archaea. AB - We have compared three complete genomes of closely related hyperthermophilic species of Archaea belonging to the Pyrococcus genus: Pyrococcus abyssi, Pyrococcus horikoshii, and Pyrococcus furiosus. At the genomic level, the comparison reveals a differential conservation among four regions of the Pyrococcus chromosomes correlated with the location of genetic elements mediating DNA reorganization. This discloses the relative contribution of the major mechanisms that promote genomic plasticity in these Archaea, namely rearrangements linked to the replication terminus, insertion sequence-mediated recombinations, and DNA integration within tRNA genes. The combination of these mechanisms leads to a high level of genomic plasticity in these hyperthermophilic Archaea, at least comparable to the plasticity observed between closely related bacteria. At the proteomic level, the comparison of the three Pyrococcus species sheds light on specific selection pressures acting both on their coding capacities and evolutionary rates. Indeed, thanks to two independent methods, the "reciprocal best hits" approach and a new distance ratio analysis, we detect the false orthology relationships within the Pyrococcus lineage. This reveals a high amount of differential gains and losses of genes since the divergence of the three closely related species. The resulting polymorphism is probably linked to an adaptation of these free-living organisms to differential environmental constraints. As a corollary, we delineate the set of orthologous genes shared by the three species, that is, the genes that may characterize the Pyrococcus genus. In this conserved core, the amino acid substitution rate is equal between P. abyssi and P. horikoshii for most of their shared proteins, even for fast evolving ones. In contrast, strong discrepancies exist among the substitution rates observed in P. furiosus relative to the two other species, which is in disagreement with the molecular clock hypothesis. PMID- 11381027 TI - Genetic evidence on the origins of Indian caste populations. AB - The origins and affinities of the approximately 1 billion people living on the subcontinent of India have long been contested. This is owing, in part, to the many different waves of immigrants that have influenced the genetic structure of India. In the most recent of these waves, Indo-European-speaking people from West Eurasia entered India from the Northwest and diffused throughout the subcontinent. They purportedly admixed with or displaced indigenous Dravidic speaking populations. Subsequently they may have established the Hindu caste system and placed themselves primarily in castes of higher rank. To explore the impact of West Eurasians on contemporary Indian caste populations, we compared mtDNA (400 bp of hypervariable region 1 and 14 restriction site polymorphisms) and Y-chromosome (20 biallelic polymorphisms and 5 short tandem repeats) variation in approximately 265 males from eight castes of different rank to approximately 750 Africans, Asians, Europeans, and other Indians. For maternally inherited mtDNA, each caste is most similar to Asians. However, 20%-30% of Indian mtDNA haplotypes belong to West Eurasian haplogroups, and the frequency of these haplotypes is proportional to caste rank, the highest frequency of West Eurasian haplotypes being found in the upper castes. In contrast, for paternally inherited Y-chromosome variation each caste is more similar to Europeans than to Asians. Moreover, the affinity to Europeans is proportionate to caste rank, the upper castes being most similar to Europeans, particularly East Europeans. These findings are consistent with greater West Eurasian male admixture with castes of higher rank. Nevertheless, the mitochondrial genome and the Y chromosome each represents only a single haploid locus and is more susceptible to large stochastic variation, bottlenecks, and selective sweeps. Thus, to increase the power of our analysis, we assayed 40 independent, biparentally inherited autosomal loci (1 LINE-1 and 39 Alu elements) in all of the caste and continental populations (approximately 600 individuals). Analysis of these data demonstrated that the upper castes have a higher affinity to Europeans than to Asians, and the upper castes are significantly more similar to Europeans than are the lower castes. Collectively, all five datasets show a trend toward upper castes being more similar to Europeans, whereas lower castes are more similar to Asians. We conclude that Indian castes are most likely to be of proto-Asian origin with West Eurasian admixture resulting in rank-related and sex-specific differences in the genetic affinities of castes to Asians and Europeans. PMID- 11381028 TI - Segmental duplications: organization and impact within the current human genome project assembly. AB - Segmental duplications play fundamental roles in both genomic disease and gene evolution. To understand their organization within the human genome, we have developed the computational tools and methods necessary to detect identity between long stretches of genomic sequence despite the presence of high copy repeats and large insertion-deletions. Here we present our analysis of the most recent genome assembly (January 2001) in which we focus on the global organization of these segments and the role they play in the whole-genome assembly process. Initially, we considered only large recent duplication events that fell well-below levels of draft sequencing error (alignments 90%-98% similar and > or =1 kb in length). Duplications (90%-98%; > or =1 kb) comprise 3.6% of all human sequence. These duplications show clustering and up to 10-fold enrichment within pericentromeric and subtelomeric regions. In terms of assembly, duplicated sequences were found to be over-represented in unordered and unassigned contigs indicating that duplicated sequences are difficult to assign to their proper position. To assess coverage of these regions within the genome, we selected BACs containing interchromosomal duplications and characterized their duplication pattern by FISH. Only 47% (106/224) of chromosomes positive by FISH had a corresponding chromosomal position by comparison. We present data that indicate that this is attributable to misassembly, misassignment, and/or decreased sequencing coverage within duplicated regions. Surprisingly, if we consider putative duplications >98% identity, we identify 10.6% (286 Mb) of the current assembly as paralogous. The majority of these alignments, we believe, represent unmerged overlaps within unique regions. Taken together the above data indicate that segmental duplications represent a significant impediment to accurate human genome assembly, requiring the development of specialized techniques to finish these exceptional regions of the genome. The identification and characterization of these highly duplicated regions represents an important step in the complete sequencing of a human reference genome. PMID- 11381029 TI - The 1.4-Mb CMT1A duplication/HNPP deletion genomic region reveals unique genome architectural features and provides insights into the recent evolution of new genes. AB - Duplication and deletion of the 1.4-Mb region in 17p12 that is delimited by two 24-kb low copy number repeats (CMT1A-REPs) represent frequent genomic rearrangements resulting in two common inherited peripheral neuropathies, Charcot Marie-Tooth disease type 1A (CMT1A) and hereditary neuropathy with liability to pressure palsy (HNPP). CMT1A and HNPP exemplify a paradigm for genomic disorders wherein unique genome architectural features result in susceptibility to DNA rearrangements that cause disease. A gene within the 1.4-Mb region, PMP22, is responsible for these disorders through a gene-dosage effect in the heterozygous duplication or deletion. However, the genomic structure of the 1.4-Mb region, including other genes contained within the rearranged genomic segment, remains essentially uncharacterized. To delineate genomic structural features, investigate higher-order genomic architecture, and identify genes in this region, we constructed PAC and BAC contigs and determined the complete nucleotide sequence. This CMT1A/HNPP genomic segment contains 1,421,129 bp of DNA. A low copy number repeat (LCR) was identified, with one copy inside and two copies outside of the 1.4-Mb region. Comparison between physical and genetic maps revealed a striking difference in recombination rates between the sexes with a lower recombination frequency in males (0.67 cM/Mb) versus females (5.5 cM/Mb). Hypothetically, this low recombination frequency in males may enable a chromosomal misalignment at proximal and distal CMT1A-REPs and promote unequal crossing over, which occurs 10 times more frequently in male meiosis. In addition to three previously described genes, five new genes (TEKT3, HS3ST3B1, NPD008/CGI 148, CDRT1, and CDRT15) and 13 predicted genes were identified. Most of these predicted genes are expressed only in embryonic stages. Analyses of the genomic region adjacent to proximal CMT1A-REP indicated an evolutionary mechanism for the formation of proximal CMT1A-REP and the creation of novel genes by DNA rearrangement during primate speciation. PMID- 11381030 TI - Comprehensive genome sequence analysis of a breast cancer amplicon. AB - Gene amplification occurs in most solid tumors and is associated with poor prognosis. Amplification of 20q13.2 is common to several tumor types including breast cancer. The 1 Mb of sequence spanning the 20q13.2 breast cancer amplicon is one of the most exhaustively studied segments of the human genome. These studies have included amplicon mapping by comparative genomic hybridization (CGH), fluorescent in-situ hybridization (FISH), array-CGH, quantitative microsatellite analysis (QUMA), and functional genomic studies. Together these studies revealed a complex amplicon structure suggesting the presence of at least two driver genes in some tumors. One of these, ZNF217, is capable of immortalizing human mammary epithelial cells (HMEC) when overexpressed. In addition, we now report the sequencing of this region in human and mouse, and on quantitative expression studies in tumors. Amplicon localization now is straightforward and the availability of human and mouse genomic sequence facilitates their functional analysis. However, comprehensive annotation of megabase-scale regions requires integration of vast amounts of information. We present a system for integrative analysis and demonstrate its utility on 1.2 Mb of sequence spanning the 20q13.2 breast cancer amplicon and 865 kb of syntenic murine sequence. We integrate tumor genome copy number measurements with exhaustive genome landscape mapping, showing that amplicon boundaries are associated with maxima in repetitive element density and a region of evolutionary instability. This integration of comprehensive sequence annotation, quantitative expression analysis, and tumor amplicon boundaries provide evidence for an additional driver gene prefoldin 4 (PFDN4), coregulated genes, conserved noncoding regions, and associate repetitive elements with regions of genomic instability at this locus. PMID- 11381031 TI - Sequence diversity in genes of lipid metabolism. AB - Elevated plasma lipoprotein levels play a crucial role in the development of coronary artery disease. Genetic factors strongly influence the levels of plasma lipoproteins, but the genes and sequence variations contributing to the most common forms of dyslipidemias are not known. We used GeneChip probe arrays to resequence the coding regions of 10 key genes of lipid metabolism. The sequences of these genes were analyzed in 80 dyslipidemic individuals. Fourteen nonsynonymous and twenty-two synonymous single nucleotide changes were identified that could be confirmed by conventional sequencing. Seven of the fourteen nonsynonymous sequence variants were polymorphisms with allele frequency >1% in the general population. The remaining seven were not found in normolipidemic controls (25 Caucasians and 25 African-Americans). The relationship between nonsynonymous sequence variations and various dyslipidemias was explored in association and family studies. No evidence was found for coding sequence variations in any of the 10 genes contributing to dyslipidemia. Only a single sequence variation, a missense mutation in the low density lipoprotein receptor gene, co-segregated with hyperlipidemia in the proband's family. This study illustrates some of the difficulties associated with identifying sequence variations contributing to complex traits. PMID- 11381032 TI - Analysis of the cat eye syndrome critical region in humans and the region of conserved synteny in mice: a search for candidate genes at or near the human chromosome 22 pericentromere. AB - We have sequenced a 1.1-Mb region of human chromosome 22q containing the dosage sensitive gene(s) responsible for cat eye syndrome (CES) as well as the 450-kb homologous region on mouse chromosome 6. Fourteen putative genes were identified within or adjacent to the human CES critical region (CESCR), including three known genes (IL-17R, ATP6E, and BID) and nine novel genes, based on EST identity. Two putative genes (CECR3 and CECR9) were identified, in the absence of EST hits, by comparing segments of human and mouse genomic sequence around two solitary amplified exons, thus showing the utility of comparative genomic sequence analysis in identifying transcripts. Of the 14 genes, 10 were confirmed to be present in the mouse genomic sequence in the same order and orientation as in human. Absent from the mouse region of conserved synteny are CECR1, a promising CES candidate gene from the center of the contig, neighboring CECR4, and CECR7 and CECR8, which are located in the gene-poor proximal 400 kb of the contig. This latter proximal region, located approximately 1 Mb from the centromere, shows abundant duplicated gene fragments typical of pericentromeric DNA. The margin of this region also delineates the boundary of conserved synteny between the CESCR and mouse chromosome 6. Because the proximal CESCR appears abundant in duplicated segments and, therefore, is likely to be gene poor, we consider the putative genes identified in the distal CESCR to represent the majority of candidate genes for involvement in CES. PMID- 11381033 TI - Sequence variability of a human pseudogene. AB - We have obtained haplotypes from the autosomal glucocerebrosidase pseudogene (psGBA) for 100 human chromosomes from worldwide populations, as well as for four chimpanzee and four gorilla chromosomes. In humans, in a 5420-nucleotide stretch analyzed, variation comprises 17 substitutions, a 3-bp deletion, and a length polymorphism at a polyadenine tract. The substitution rate on the pseudogene (1.23 +/- 0.22 x 10(-9) per nucleotide and year) is within the range of previous estimates considering phylogenetic estimations. Recombination within the pseudogene was recognized, although the low variability of this locus prevented an accurate measure of recombination rates. At least 13% of the psGBA sequence could be attributed to gene conversion from the contiguous GBA gene, whereas the reciprocal event has been shown to lead to Gaucher disease. Human psGBA sequences showed a recent coalescence time (approximately 200,000 yr ago), and the most ancestral haplotype was found only in Africans; both observations are compatible with the replacement hypothesis of human origins. In a deeper timeframe, phylogenetic analysis showed that the duplication event that created psGBA could be dated at approximately 27 million years ago, in agreement with previous estimates. PMID- 11381034 TI - Sequence-based design of single-copy genomic DNA probes for fluorescence in situ hybridization. AB - Chromosomal rearrangements are frequently monitored by fluorescence in situ hybridization (FISH) using large, recombinant DNA probes consisting of contiguous genomic intervals that are often distant from disease loci. We developed smaller, targeted, single-copy probes directly from the human genome sequence. These single-copy FISH (scFISH) probes were designed by computational sequence analysis of approximately 100-kb genomic sequences. ScFISH probes are produced by long PCR, then purified, labeled, and hybridized individually or in combination to human chromosomes. Preannealing or blocking with unlabeled, repetitive DNA is unnecessary, as scFISH probes lack repetitive DNA sequences. The hybridization results are analogous to conventional FISH, except that shorter probes can be readily visualized. Combinations of probes from the same region gave single hybridization signals on metaphase chromosomes. ScFISH probes are produced directly from genomic DNA, and thus more quickly than by recombinant DNA techniques. We developed single-copy probes for three chromosomal regions-the CDC2L1 (chromosome 1p36), MAGEL2 (chromosome 15q11.2), and HIRA (chromosome 22q11.2) genes-and show their utility for FISH. The smallest probe tested was 2290 bp in length. To assess the potential utility of scFISH for high-resolution analysis, we determined chromosomal distributions of such probes. Single-copy intervals of this length or greater are separated by an average of 29.2 and 22.3 kb on chromosomes 21 and 22, respectively. This indicates that abnormalities seen on metaphase chromosomes could be characterized with scFISH probes at a resolution greater than previously possible. PMID- 11381035 TI - Rapid amplification of plasmid and phage DNA using Phi 29 DNA polymerase and multiply-primed rolling circle amplification. AB - We describe a simple method of using rolling circle amplification to amplify vector DNA such as M13 or plasmid DNA from single colonies or plaques. Using random primers and phi29 DNA polymerase, circular DNA templates can be amplified 10,000-fold in a few hours. This procedure removes the need for lengthy growth periods and traditional DNA isolation methods. Reaction products can be used directly for DNA sequencing after phosphatase treatment to inactivate unincorporated nucleotides. Amplified products can also be used for in vitro cloning, library construction, and other molecular biology applications. PMID- 11381036 TI - Single nucleotide polymorphism markers for genetic mapping in Drosophila melanogaster. AB - For nearly a century, genetic analysis in Drosophila melanogaster has been a powerful tool for analyzing gene function, yet Drosophila lacks the molecular genetic mapping tools that recently have revolutionized human, mouse, and plant genetics. Here, we describe the systematic characterization of a dense set of molecular markers in Drosophila by using a sequence tagged site-based physical map of the genome. We identify 474 biallelic markers in standard laboratory strains of Drosophila that span the genome. Most of these markers are single nucleotide polymorphisms and sequences for these variants are provided in an accessible format. The average density of the new markers is one per 225 kb on the autosomes and one per megabase on the X chromosome. We include in this survey a set of P-element strains that provide additional use for high-resolution mapping. We show one application of the new markers in a simple set of crosses to map a mutation in the hedgehog gene to an interval of <1 Mb. This new map resource significantly increases the efficiency and resolution of recombination mapping and will be of immediate value to the Drosophila research community. PMID- 11381037 TI - A systematic analysis of human disease-associated gene sequences in Drosophila melanogaster. AB - We performed a systematic analysis of 929 human disease gene entries associated with at least one mutant allele in the Online Mendelian Inheritance in Man (OMIM) database against the recently completed genome sequence of Drosophila melanogaster. The results of this search have been formatted as an updateable and searchable on-line database called Homophila. Our analysis identified 714 distinct human disease genes (77% of disease genes searched) matching 548 unique Drosophila sequences, which we have summarized by disease category. This breakdown into disease classes creates a picture of disease genes that are amenable to study using Drosophila as the model organism. Of the 548 Drosophila genes related to human disease genes, 153 are associated with known mutant alleles and 56 more are tagged by P-element insertions in or near the gene. Examples of how to use the database to identify Drosophila genes related to human disease genes are presented. We anticipate that cross-genomic analysis of human disease genes using the power of Drosophila second-site modifier screens will promote interaction between human and Drosophila research groups, accelerating the understanding of the pathogenesis of human genetic disease. The Homophila database is available at http://homophila.sdsc.edu. PMID- 11381039 TI - Small samples: does size matter? PMID- 11381040 TI - Tumor-infiltrating macrophages (CD68(+) cells) and prognosis in malignant uveal melanoma. AB - PURPOSE: To investigate the hypothesis that tumor-infiltrating macrophages contribute to prognosis of uveal melanoma and to study their association with tumor characteristics, especially microvessels. METHODS: This was a retrospective, population-based cohort study of 167 consecutive patients who had had an eye with choroidal and ciliary body melanoma removed between 1972 and 1981. Macrophages were identified with mAb PG-M1 to the CD68 epitope, and their number and morphologic type were recorded. Kaplan-Meier and Cox regression analyses of melanoma-specific survival were performed. RESULTS: CD68-positive macrophages could be assessed in 139 (83%) of the 167 melanomas. Their number was moderate to high in 115 (83%) of the 139 tumors, and their morphology ranged from dendritic to round. A high number of macrophages was associated with presence of epithelioid cells (P = 0.025), heavy pigmentation (P = 0.001), and high microvascular density (P = 0.001). The 10-year melanoma-specific mortality rate increased with higher numbers of macrophages (0.10 for low versus 0.57 for high numbers, P = 0.0012). The morphologic type of infiltrating macrophages was not associated with mortality. The number of macrophages was modeled by stratification, which significantly improved a Cox regression model (P < 0.001). Adjusting for the other independent indicators of metastatic death 10-year melanoma-specific mortality was 0.17 for low versus 0.45 for high numbers of macrophages. CONCLUSIONS: The number of tumor-infiltrating CD68-positive macrophages contributes to prognosis and associates with cell type and microvascular density, which merits a further analysis of the biological role of these cells in uveal melanoma. PMID- 11381038 TI - Neuropeptides and neuropeptide receptors in the Drosophila melanogaster genome. AB - Recent genetic analyses in worms, flies, and mammals illustrate the importance of bioactive peptides in controlling numerous complex behaviors, such as feeding and circadian locomotion. To pursue a comprehensive genetic analysis of bioactive peptide signaling, we have scanned the recently completed Drosophila genome sequence for G protein-coupled receptors sensitive to bioactive peptides (peptide GPCRs). Here we describe 44 genes that represent the vast majority, and perhaps all, of the peptide GPCRs encoded in the fly genome. We also scanned for genes encoding potential ligands and describe 22 bioactive peptide precursors. At least 32 Drosophila peptide receptors appear to have evolved from common ancestors of 15 monophyletic vertebrate GPCR subgroups (e.g., the ancestral gastrin/cholecystokinin receptor). Six pairs of receptors are paralogs, representing recent gene duplications. Together, these findings shed light on the evolutionary history of peptide GPCRs, and they provide a template for physiological and genetic analyses of peptide signaling in Drosophila. PMID- 11381041 TI - Lack of lymphangiogenesis despite coexpression of VEGF-C and its receptor Flt-4 in uveal melanoma. AB - PURPOSE: Because lymphatic vessels are absent from the normal eye and because uveal melanomas are presumed to spread by a hematogenous route in the absence of tumor exposure to conjunctival lymphatics, this study was undertaken to investigate the presence of lymphatic vessels in primary uveal melanomas. METHODS: The presence of lymphatics in 2 control eyes and in 33 primary uveal, 10 primary cutaneous, and 3 metastatic cutaneous melanomas was evaluated by using a double-immunostaining protocol that differentially highlights blood and lymphatic vasculature. In addition, 14 uveal melanomas were immunostained for the lymphatic growth factor vascular endothelial growth factor (VEGF)-C (with anti-VEGF-C polyclonal antibodies [pAbs]), its receptors Flt-4 (with monoclonal antibody [mAb] 9D9) and KDR (with anti-KDR mAb [Clone KDR-2]), and the hemangiogenic factor VEGF-A (with anti-VEGF pAbs). RESULTS: Lymphatics were not detected in normal eyes or in uveal melanoma. As a consequence, signs of lymphangiogenesis were not present. There was coexpression of VEGF-C with Flt-4 and KDR in 6 (43%) of the 14 melanomas. Staining for VEGF-A was completely negative in 25 uveal melanomas analyzed. CONCLUSIONS: The strictly hematogenous metastasis of primary uveal melanomas is explained by the absence of lymphatics in and around the tumor. The current data suggest that, in the presence of endothelial Flt-4, VEGF C expression is not sufficient to induce lymphangiogenesis from preexisting blood vessels in human cancer. PMID- 11381042 TI - Expression, purification, and MALDI analysis of RPE65. AB - PURPOSE: RPE65 is preferentially expressed in the retinal pigment epithelium (RPE) and is essential for retinal function. The purpose of the study was to develop methods for the expression of the protein, determine the accurate molecular weight of this expressed protein, and quantitate the amount of RPE65 in the bovine RPE. METHODS: Human RPE65 was expressed in Sf9 cells using the baculovirus system. The subcellular localization was determined by Western blot analysis and immunocytochemistry. An ELISA was developed for RPE65 and used to measure levels in bovine RPE. Recombinant and native RPE65 were purified by affinity chromatography. Molecular mass was determined by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. RESULTS: Recombinant human (rH)RPE65 was expressed as a major protein associated with cell membrane in Sf9 cells. The recombinant protein was purified to apparent homogeneity from both the membrane and nonmembrane fractions. The identity of the purified protein was confirmed by Western blot analysis and by partial peptide sequencing. rHRPE65 from the nonmembrane fraction has a mass of 64,867 +/- 80 which is close to the calculated molecular weight from the amino acid sequence including the His-tag (64,663), whereas the membrane-associated rHRPE65 has a molecular mass of 65,380 +/- 150, which is significantly higher than that of the non-membrane-associated form and the calculated molecular weight, suggesting posttranslational modifications. Similarly, native RPE65 was detected in the cytosolic and microsomal fractions of the bovine RPE, with an average level of 3.8 +/- 1.3 and 7.2 +/- 0.4 microg RPE65 per eye, respectively. The cytosolic form had a molecular mass of 61,161 +/- 60, which is close to the calculated value (60,944), whereas that of the microsomal form was 61,961 +/- 170. CONCLUSIONS: RPE65 is expressed in two forms, one of which is membrane associated and contains significant posttranslational modifications, similar to the native membrane associated form. PMID- 11381043 TI - A new locus for autosomal recessive RP (RP29) mapping to chromosome 4q32-q34 in a Pakistani family. AB - PURPOSE: To map the disease locus in a six-generation, consanguineous Pakistani family with autosomal recessive retinitis pigmentosa (arRP). All affected individuals had pigmentary retinopathy associated with symptoms of night blindness and the loss of peripheral visual fields by the age of 20 years, loss of central vision between the ages of 25 and 30 years, and complete blindness between the ages of 40 and 50 years. METHODS: Genomic DNA from family members was typed for alleles at known polymorphic genetic markers using polymerase chain reaction. Alleles were assigned to individuals, which allowed calculation of LOD scores using the programs Cyrillic (http://www.cyrillicsoftware.com) and MLINK (Cherwell Scientific Publishing LTD:, Oxford, UK). The genes for membrane glycoprotein (M6a) and chloride channel 3 (CLCN3) were analyzed by direct sequencing for mutations. RESULTS: A new locus for arRP (RP29) has been mapped to chromosome 4q32-q34. A maximum two-point LOD score of 3.76 was obtained for the marker D4S415, with no recombination. Two recombination events in the pedigree positioned this locus to a region flanked by markers D4S621 and D4S2417. A putative region of homozygosity by descent was observed between the loci D4S3035 and D4S2417, giving a probable disease interval of 4.6 cM. Mutation screening of two candidate genes, M6a and CLCN3, revealed no disease-associated mutations. CONCLUSIONS: The results suggest that the arRP phenotype maps to a new locus and is due to a mutated gene within the 4q32-q34 chromosomal region. PMID- 11381044 TI - Activation of arrestin: requirement of phosphorylation as the negative charge on residues in synthetic peptides from the carboxyl-terminal region of rhodopsin. AB - PURPOSE: To determine whether substitution of the potential phosphorylation sites of bovine rhodopsin's carboxyl-terminal region with the acidic residues aspartic acid, glutamic acid, or cysteic acid promotes the activation of arrestin. METHODS: Three peptide analogues of the 19-residue carboxyl-terminal region of rhodopsin (330-348) were synthesized: the fully phosphorylated peptide (7P peptide), the peptide with all potential phosphorylation sites substituted with glutamic acid (7E-peptide), and the peptide with the phosphorylation sites substituted with cysteic acid (7Cya-peptide). The peptides were tested in assays in which the 7P-peptide had previously been shown to have an effect. Rhodopsin with glutamic acid (Etail) or aspartic acid (Dtail) substituted for the phosphorylation sites in rhodopsin were constructed and expressed in COS-7 cells and tested in an in vitro assay. RESULTS: Earlier work has demonstrated that the 7P-peptide activates arrestin, showing induction of arrestin binding to light activated unphosphorylated rhodopsin, inhibition of the light-induced phosphodiesterase (PDE) activity in rod outer segments (ROS) with excess arrestin, increase in the initial rapid proteolysis of arrestin by trypsin, and enhanced reactivity of one of arrestin's sulfhydryl groups with inhibition of the reactivity of another. None of these effects was observed in the presence of 7E peptide or 7Cya-peptide. The 7Cya-peptide inhibited the PDE activity in ROS, but the same effect was observed both in the presence and the absence of excess arrestin. Because none of the other effects was observed with the 7Cya-peptide, the authors conclude that the 7Cya-peptide does not activate arrestin, but acts, probably nonspecifically, through some other part of the transduction system. Considerable arrestin-mediated rhodopsin inactivation was observed with both the Etail and the Dtail mutant, although these substitutions did not yield rhodopsins that were equivalent to phosphorylated rhodopsin. CONCLUSIONS: These results, taken together, suggest that the negative charge due to phosphates in the carboxyl-terminal region of rhodopsin are required for the full activation of arrestin and that acidic amino acids (carboxyl and sulfonic) do not mimic the negative charge of phosphorylated residues. PMID- 11381045 TI - Measuring and interpreting the incidence of congenital ocular anomalies: lessons from a national study of congenital cataract in the UK. AB - PURPOSE: Prevention of visual impairment due to congenital cataract is an international priority. Estimates of incidence are required for implementation and assessment of preventive strategies, but are not widely available, despite routine monitoring of birth defects at a national level in many industrialized countries. The purpose of this study was to determine the incidence of new diagnosis of congenital and infantile cataract in the United Kingdom. METHODS: All children with newly diagnosed congenital and infantile cataract in the United Kingdom in 1 year from October 1995 through September 1996 were identified using independent ophthalmic and pediatric national active surveillance schemes. Capture-recapture analysis was used to estimate completeness of ascertainment. Annual age-specific and cumulative incidence were estimated and adjusted for ascertainment. RESULTS: Two hundred forty-eight children with newly diagnosed congenital or infantile cataract were identified-an estimated 92% of eligible cases. The adjusted annual age-specific incidence of new diagnosis of congenital and infantile cataract was highest in the first year of life, being 2.49 per 10,000 children (95% confidence interval [CI], 2.10-2.87). Adjusted cumulative incidence at 5 years was 3.18 per 10,000 (95% CI, 2.76-3.59), increasing to 3.46 per 10,000 by 15 years (95% CI, 3.02-3.90). Incidence of bilateral cataract was higher than that of unilateral, but incidence did not vary by sex or country of residence. CONCLUSIONS: These estimates of congenital and infantile cataract incidence were higher than reported previously from routine sources relying on passive notification around the time of birth. Studies of congenital ocular anomalies that are not always readily diagnosed at birth should consider the potential influence on disease frequency of diagnostic practices as well as of underlying disease risk. PMID- 11381046 TI - Refractive errors and incident cataracts: the Beaver Dam Eye Study. AB - PURPOSE: To describe the relation between refractive errors and incident age related cataracts in a predominantly white US population. METHODS: All persons aged 43 to 84 years of age in Beaver Dam, Wisconsin, were invited for a baseline examination from 1988 through 1990 and a follow-up examination 5 years later from 1993 through 1995. At both examinations, participants had refraction and photographic assessment of cataract, according to a standardized protocol. Myopia was defined as a spherical equivalent of -1.0 diopters (D) or less, hyperopia as +1.0 D or more. The relations between refractive errors at baseline and cataract at baseline (prevalent cataract), 5-year incident cataract, and incident cataract surgery were analyzed by using generalized estimating equations. RESULTS: When age and gender were controlled for, myopia was related to prevalent nuclear cataract (odds ratio [OR], 1.67; 95% confidence interval [CI], 1.23-2.27), but not to cortical and posterior subcapsular cataracts. Myopia was not related to 5 year incident nuclear, cortical, and posterior subcapsular cataracts, but was related to incident cataract surgery (OR 1.89; CI 1.18-3.04). Hyperopia was related to incident nuclear (OR 1.56; CI 1.25-1.95) and possibly cortical (OR 1.25; CI 0.96-1.63) cataracts, but not to posterior subcapsular cataract or cataract surgery. After further adjustment for diabetes, smoking, and education, the association between myopia and incident cataract surgery was attenuated (OR 1.60; CI 0.96-2.64), but the associations between hyperopia and incident nuclear and cortical cataracts were unchanged. CONCLUSIONS: These data support the cross sectional association between myopia and nuclear cataract seen in other population-based studies, but provide no evidence of a relationship between myopia and 5-year incident cataract. Hyperopia may be related weakly to incident nuclear and cortical cataract. PMID- 11381047 TI - Isolation, characterization, and propagation of rat conjunctival goblet cells in vitro. AB - PURPOSE: To isolate, culture, and characterize goblet cells from the conjunctiva of rats. METHODS: Conjunctival tissue was surgically removed from Sprague-Dawley rats. Goblet cells were then isolated from the nictitating membrane and fornix using explant cultures. Cells derived from the explants were grown and propagated in RPMI medium supplemented with 10% fetal bovine serum. They were characterized using an enzyme-linked lectin assay (ELLA) with the lectin Ulex europaeus agglutinin-1 (UEA-1), Western blot analysis, PCR, light and electron microscopy, specialized histochemistry and indirect immunofluorescence microscopy. RESULTS: Goblet cells were successfully isolated from conjunctival explants by scraping nongoblet cells from the culture vessel. To date, cultures have been passaged a minimum of three times without the loss of their specific cellular markers. Cells identified as goblet cells fulfilled the following criteria: positive staining for alcian blue/periodic acid Schiff reagent, cytokeratin (CK)-7, the lectins UEA I and Helix pomatia agglutinin (HPA), MUC5AC, and M(3) muscarinic receptor; detection of MUC5AC mRNA using RT-PCR; and negative staining for CK-4, M(1) muscarinic receptor, and Banderia simplicifolia lectin. The authors also measured, using the ELLA, substantial amounts of UEA-I-detectable high-molecular weight glycoproteins and MUC5AC released into the medium. CONCLUSIONS: Cultured goblet cells retain many characteristics of goblet cells in vivo and thus may serve as a useful tool in delineating the pathobiology of the ocular surface. PMID- 11381048 TI - TGF-beta receptor types I and II are differentially expressed during corneal epithelial wound repair. AB - PURPOSE: It has been demonstrated that cells migrating to cover an epithelial debridement wound exit the cell cycle and that the cell-cycle inhibitor p15(INK4b) is upregulated in these cells. TGF-beta signaling has been implicated in both of these processes, and this study was conducted to determine whether the expression and localization of TGF-beta receptor (TbetaR)-I and -II are altered during corneal epithelial wound repair. METHODS: Three-millimeter superficial keratectomy wounds and 3-mm debridement wounds were made in central rat cornea and allowed to heal in vivo for 1 to 48 hours. Immunofluorescence microscopy and Western blot analysis were used to determine the localization and expression of TbetaR-I and -II. Unwounded rat corneas served as control samples. To determine the effect of epidermal growth factor (EGF) and TGF-beta1 on p15(INK4b) and TbetaR-I and -II expression, human corneal epithelial cells were grown in culture to 50% to 60% confluence, and EGF (5 ng/ml) and/or TGF-beta1 (2 ng/ml) were added for 6 hours. Cells were harvested and p15(INK4b) and TBR-I and -II levels were assayed by using Western blot analysis. RESULTS: In unwounded corneas, TbetaR-I and TbetaR-II were present at low levels across the cornea, with higher levels in limbal epithelium. Both TbetaR-I and -II were upregulated after wounding. However, levels of TbetaR-II appeared to increase in the epithelial cells that had migrated to cover the wound area, whereas TbetaR-I was upregulated in the entire corneal epithelium. Western blot analysis indicated that both TbetaR-I and -II were upregulated threefold after wounding. In cultured cells, EGF and TGF beta1 stimulated TbetaR-II; however, neither one stimulated TbetaR-I expression. TGF-beta1 stimulated p15(INK4b) protein levels threefold. CONCLUSIONS: After wounding, TbetaR-I and TbetaR-II were both expressed at high levels in cells migrating to cover a corneal wound, suggesting that TGF-beta signaling is involved in blocking migrating cells from progressing through the cell cycle. This blockage, at least in part, involves the inhibitor p15(INK4b). In addition, although both TbetaR-I and TbetaR-II are upregulated during wound repair, they appear to be differentially regulated. PMID- 11381049 TI - Upregulation of phospholipase Cgamma1 activity during EGF-induced proliferation of corneal epithelial cells: effect of phosphoinositide-3 kinase. AB - PURPOSE: Previously, the authors showed that epidermal growth factor (EGF) stimulates phospholipase Cgamma1 (PLCgamma1) and phosphoinositide-3 kinase (PI3K) activities in confluent rabbit corneal epithelial cells (RCECs). The purpose of this study was to investigate whether PLCgamma1 activity is upregulated during EGF-induced proliferation of RCECs and to determine whether there is any cross talk between PLCgamma1 and PI3K in these cells. METHODS: Simian virus (SV)-40 immortalized RCECs were cultured in the presence and absence of EGF and other agents. At prescribed time intervals, the cultures were terminated and the cells counted. PLCgamma1 activity in intact cells was assessed by measuring the production of [(3)H]IP(3) in [(3)H]myoinositol-labeled cells. The in vitro enzyme activity was assayed using immunoprecipitated PLCgamma1 and [(3)H]PI(4,5)P(2) as substrate. [(3)H]IP(3), the product of PLCgamma1, was analyzed by anion-exchange chromatography. The changes in protein content and level of phosphorylation of PLCgamma1 were determined by Western immunoblot analysis, with the appropriate antibodies. RESULTS: Addition of EGF (50 ng/ml) caused a time-dependent increase in proliferation of RCECS. The effect of EGF peaked at approximately 36 hours. Under the same experimental conditions, EGF stimulated PLCgamma1 activity with a time course similar to that of cell proliferation. Data from Western immunoblot analysis revealed that the EGF-stimulated PLCgamma1 activity was due to increased synthesis of the enzyme. Furthermore, during cell proliferation, tyrosine phosphorylation of PLCgamma1 increased in a time-dependent manner that corresponded closely with the expression of PLCgamma1. EGF exerted its effects both on cell proliferation and PLCgamma1 activation in a dose-dependent manner. Treatment of the cells with U-73122, a PLC inhibitor, or myr-GLYRKAMRLRY, a myristoylated PLCgamma1 inhibitor peptide, caused attenuation of both the EGF stimulated cell proliferation and PLCgamma1 activity. Treatment of the cells with the PI3K inhibitors, wortmannin or LY294002, caused inhibition of both EGF stimulated cell proliferation and PLCgamma1 activation. Addition of PI(3,4,5)P(3) to the in vitro PLCgamma1 assay mixture stimulated the enzyme activity in a dose dependent manner. CONCLUSIONS: The data suggest a positive correlation between EGF-stimulated PLCgamma1 activation and cell proliferation in RCECS. The EGF stimulated PLCgamma1 activity was mirrored by increased synthesis and tyrosine phosphorylation of the enzyme. The data also show that PLCgamma1 activation and cell proliferation were inhibited by PI3K inhibitors, suggesting a role for PI3K in EGF-stimulated proliferation of corneal epithelial cells. PMID- 11381050 TI - Adaptive changes in dynamic properties of human disparity-induced vergence. AB - PURPOSE: Vergence eye movements undergo adaptive recalibration in response to a training stimulus in which the initial disparity is changed just after vergence begins (the double-step paradigm). In the present study the changes in the dynamic properties of convergence, speed and acceleration, were examined by using this double-step paradigm, before and after adaptation. METHODS: Four normal subjects participated. Three-dimensional visual stimuli were provided by a head mounted display with two liquid crystal diode (LCD) panels. To induce adaptation, a double step of disparity was used: an initial step from distances of 2 to 1 m was followed by a second step to distances of 0.7 m ("increasing paradigm") or 1.4 m ("decreasing paradigm") after a constant period of 0.2 seconds. The dynamic properties of vergence were compared before and after 30 minutes of training with these paradigms. RESULTS: Peak velocity of convergence became significantly greater (increasing paradigm) or smaller (decreasing paradigm) after 30 minutes' training. Changes in the dynamic properties of convergence were also obvious in phase-plane (velocity versus position) and main sequence (peak velocity versus amplitude) plots. Further analysis revealed that adaptive increases in vergence velocity were accomplished by an increase in the duration of the acceleration period, whereas adaptive decreases were induced by a decrease in the maximum value of acceleration. CONCLUSIONS: The pattern of change in the dynamic characteristics of vergence after adaptation was similar to that of saccades and the initiation of pursuit eye movements, suggesting common neural mechanisms for adaptive changes in the open-loop control of eye movements. PMID- 11381051 TI - Correlation of optic nerve head tomography with visual field sensitivity in papilledema. AB - PURPOSE: To quantify the relationship between optic nerve head tomography and perimetric sensitivity in patients with papilledema. METHODS: Eight patients with variable degrees of recently diagnosed papilledema associated with idiopathic intracranial hypertension (IIH) were evaluated with confocal scanning laser ophthalmoscopy (CSLO) and automated perimetry. Patients were followed up with serial measurements over a period of 5 to 30 months (mean +/- SD, 17.1 +/- 9), while under medical treatment (acetazolamide). The tomographic parameters, volume above reference (VAR), volume above surface (VAS), effective mean height (EMH), and maximum height in contour (MxHC), were obtained by tomography, either globally or from predefined disc sectors. The perimetric indices, mean deviation (MD) and pattern SD (PSD), were evaluated. The results from patients' right eyes and the individual intereye differences in both tomographic and perimetric parameters, were statistically evaluated by nonparametric correlational (Spearman) and repeated measures (Wilcoxon) analyses. RESULTS: At baseline, all tomographic parameters were negatively correlated with MD in global (r = -0.8) and sectorial (r = -0.6) evaluations. The interocular differences in overall tomographic parameters were correlated with corresponding differences in perimetric MD (r = -0.8) and PSD (r = 0.6). During the follow-up period, volumetric disc parameters decreased (P < 0.02), whereas perimetric MD increased (P = 0.02) at comparable times. CONCLUSIONS: In patients with recently diagnosed papilledema, optic nerve head tomographic abnormalities are quantitatively correlated with visual field sensitivity losses. Therapeutic improvement of volumetric parameters may be paralleled by recovery in perimetric sensitivity. The data support the possible use of both techniques in combination to monitor the amount of papilledema and the effectiveness of treatments designed to reduce intracranial hypertension. PMID- 11381052 TI - Spatial localization after different types of retinal detachment surgery. AB - PURPOSE: To compare the effect on spatial localization of two different forms of surgery for primary rhegmatogenous retinal detachment. METHODS: Two groups of 30 patients (one group undergoing conventional external scleral-buckling procedures, the other undergoing vitrectomy procedures) were recruited. They pointed at targets appearing on a computer touchscreen without being able to see their hands, while viewing targets with the non-surgically treated eye. The sizes of the horizontal pointing errors were recorded on three separate occasions: before surgery, on the first postoperative day, and approximately 10 days later. RESULTS: On the first postoperative day a significant change in localization of 2.9 +/- 0.9 degrees [SD]) was observed in the scleral-buckling group, compared with 1.3 +/- 0.6 degrees in the vitrectomy group. These changes resolved by the second postoperative assessment. CONCLUSIONS: These results, particularly in patients in the scleral-buckling group in whom greater manipulation of the extraocular muscles inevitably occurs, are consistent with an alteration in the extraretinal eye position information that is used in spatial localization. This is likely to be a consequence of modified efference copy and/or extraocular muscle proprioception. PMID- 11381053 TI - Comparison of the vasoactive effects of the docosanoid unoprostone and selected prostanoids on isolated perfused retinal arterioles. AB - PURPOSE: To compare the vasoactive properties of the docosanoid unoprostone, its free acid, and different members of the prostanoid family on isolated perfused pig retinal arterioles to assess their potential to modulate retinal blood flow. METHODS: Segments of porcine retinal arterioles were dissected, cannulated, and perfused, and their diameter monitored during either intraluminal or extraluminal application of increasing doses (10(-10)-10(-4) M) of either the docosanoid unoprostone isopropyl and its free acid or of selected prostanoids: prostaglandin (PG) F(2alpha) and thromboxane A(2) analogue (U46619). Studies were performed on arterioles in their uncontracted state, and also during precontraction with endothelin-1 (10(-9) M). The significance of any induced change in vessel diameter was assessed in relation to the initial vessel diameter or, in the case of endothelin-1 administration, to the contracted diameter with endothelin-1 alone. RESULTS: In normal-tone arterioles without endothelin-1 contraction, PGF(2alpha) and U46619 both produced a potent dose-dependent contraction, but neither unoprostone isopropyl nor unoprostone free acid had a significant vasoactive effect. In endothelin-1-contracted arterioles, U46619 produced further contraction, PGF(2alpha) produced a slight vasodilatation, and unoprostone isopropyl and its free acid produced a pronounced dilatation. CONCLUSIONS: Of the agents tested, unoprostone isopropyl and its free acid were the most potent vasodilators of endothelin-1-contracted pig retinal arterioles. Members of the prostanoid family demonstrated a different effect on the diameter of isolated retinal arterioles compared with the docosanoids. The potential therefore exists for the docosanoid unoprostone to have a beneficial effect on retinal blood flow in addition to any reduction in intraocular pressure. PMID- 11381054 TI - Effects of mechanical stretching on trabecular matrix metalloproteinases. AB - PURPOSE: The homeostatic mechanisms responsible for intraocular pressure (IOP) regulation are not understood. Studies were conducted to evaluate the hypothesis that trabecular meshwork (TM) cells sense increases in IOP as stretching or distortion of their extracellular matrix (ECM) and respond by increasing ECM turnover enzymes. METHODS: Flow rates were increased in perfused human anterior segment organ cultures and the matrix metalloproteinase (MMP) levels and IOP were evaluated. Human TMs in stationary anterior segment organ culture were mechanically stretched, and MMP levels were analyzed. TM cells were grown on membranes, which were then stretched, and MMP levels were evaluated. Western immunoblots, zymography, and confocal immunohistochemistry were used to evaluate changes in MMPs and their tissue inhibitors, the TIMPS: RESULTS: Doubling the flow rate in perfused human organ cultures increased gelatinase A levels in the perfusate by 30% to 50% without affecting gelatinase B or stromelysin levels. Immediately after doubling the flow rate, the measured IOP doubled. However, over the next few days the IOP gradually returned to the initial level, although the flow rate was maintained at double the initial value. Stretching stationary organ cultures or stretching TM cells grown on membranes resulted in similar increases in gelatinase A without changes in gelatinase B or stromelysin levels. The gelatinase A increases occurred between 24 and 72 hours and were approximately proportional to the degree of stretching. Although coating the membranes with different ECM molecule affected the gelatinase A response, the optimum response occurred when the cells had been grown long enough to produce their own ECM. By Western immunoblot and confocal immunohistochemistry, the stretch-induced increases in gelatinase A were accompanied by strong decreases in TIMP-2 levels and moderate increases in one membrane type MMP, MT1-MMP. After mechanical stretching of the membrane, gelatinase A, MT1-MMP and TIMP-2 all exhibited a similar punctate immunostaining pattern over the TM cell surface. CONCLUSIONS: These results are compatible with the hypothesis that elevations in IOP are sensed by TM cells as ECM stretch/distortion. TM cells respond by increasing gelatinase A and MT1-MMP, while decreasing TIMP-2 levels. This will increase ECM turnover rates, reduce the trabecular resistance to aqueous humor outflow, and restore normal IOP levels. This hypothesis provides a regulatory feedback mechanism for IOP homeostasis. PMID- 11381055 TI - Increased human scleral permeability with prostaglandin exposure. AB - PURPOSE: To investigate the effect of prostaglandins (PGs) on the permeability of human sclera in vitro. METHODS: Twenty-three pairs of human eye bank eyes were studied. Circular pieces of sclera were cultured in low-serum DMEM/F-12 media. Scleral hydration was assessed by measuring wet and dry weight of scleral cultures incubated with medium for 3 days and with Hanks' buffered saline solution (HBSS) for 4 hours. To assess scleral permeability, organ-cultured scleral tissues were exposed to 100 to 500 nM PGF(2alpha), 17-phenyltrinor PGF(2alpha), or PhXA85 (the active form of latanoprost) for 1, 2, and 3 days. Scleral permeability was measured using a two-chamber Ussing apparatus and rhodamine-dextran polymers dissolved in HBSS (MW = 10,000, 40,000, and 70,000). The movement of each rhodamine-dextran across the cultured sclera was measured using a spectrofluorometer. To understand the biological basis of the permeability change, the media were collected from the treated cultures, and the concentration of MMP-1, 2, and 3 was measured using an enzyme-linked immunosorbent assay. RESULTS: There was no difference in scleral hydration among fresh sclera and sclera incubated with medium for 3 days, with HBSS for 4 hours, or with medium for 3 days followed by HBSS for 4 hours. Compared to tracer movement across untreated scleral cultures (1.5 x 10(-6) cm/sec for 10 kDa dextran, 0.7 x 10(-6) cm/sec for 40 kDa dextran, and 0.4 x 10(-6) cm/sec for 70 kDa dextran), exposure to PGF(2alpha), 17-phenyltrinor PGF(2alpha), or PhXA85 each increased scleral permeability in a dose- and time-dependent manner. Increases in permeability were greater with the10 kDa dextran than with the 40 or 70 kDa dextran. The magnitude of these effects was greatest with exposure to PhXA85 and similar with exposure to PGF(2alpha) or 17-phenyltrinor-PGF(2alpha). MMP expression also was significantly increased after PG exposure. These increases were generally time and dose dependent and greater with MMP-2 and -3 than with MMP-1. CONCLUSIONS: There is increased permeability of human sclera exposed to various PGs in organ culture. This increased permeability is accompanied by increased expression of MMPS: PMID- 11381056 TI - Induction of heat shock protein 72 protects retinal ganglion cells in a rat glaucoma model. AB - PURPOSE: To investigate whether heat shock protein (Hsp) 72 is induced in retinal ganglion cells (RGCs) in experimental rat glaucoma and whether the induction of Hsp72 by heat stress or zinc (Zn(2+)) administration can increase survival of RGCs in the model. METHODS: Intraocular pressure (IOP) was elevated unilaterally in Wistar rats with argon laser irradiation of the trabecular meshwork 5 days after intracameral injection of india ink. Immunohistochemical staining for Hsp72 was performed. The rats with elevated IOP were treated with heat stress once a week (six rats) or intraperitoneal injection of zinc (10 mg/kg) every two weeks (six rats). Untreated rats with elevated IOP served as a control group (six rats). Quercetin, an inhibitor of Hsp expression was injected in the rats with heat stress (six rats) and zinc injection (seven rats). Subsequent to 4 weeks of IOP elevation, RGCs were counted. RESULTS: The IOP increase compared with the contralateral eyes was 48% +/- 4% throughout the study period. Hsp72 was detected only in the eyes with elevated IOP at 1 and 2 days and was weakly detected at 1 week of IOP elevation. A single administration of zinc strongly induced Hsp72 in RGCs of rats with elevated IOP for 2 weeks. Treatment with heat stress or zinc in rats with elevated IOP increased RGC survival after 4 weeks of IOP elevation, compared with the untreated control group (P = 0.004, n = 6). Quercetin reversed the positive effect of heat stress or zinc injection on RGC survival. CONCLUSIONS: These results demonstrate the possibility of a novel therapeutic approach to glaucoma through an enhanced induction of the endogenous heat shock response. PMID- 11381057 TI - Human tenon's fibroblast-produced ifnbeta and the prevention of t-cell apoptosis. AB - PURPOSE: Fibroblast-T-cell interactions may contribute to the development of chronic inflammation, a risk factor for trabeculectomy failure. This study was undertaken to determine whether normal and growth-arrested human Tenon's fibroblasts (HTF) can prevent cytokine deprivation-mediated T-cell apoptosis through the secretion of interferon (IFN)beta. METHODS: HTF were used either untreated or pretreated with mitomycin-C (MMC; 0.1 or 0.4 mg/ml) or 5 fluorouracil (5FU; 25 or 50 mg/ml). IL2-deprived T cells were cocultured with HTF. T-cell viability was measured at specific time points. Human Tenon's fibroblast-conditioned medium was used either untreated or treated with a neutralizing antibody against IFNbeta to block its action, after which IL2 deprived T cells were added and T-cell viability was measured. An image analysis system was used to determine the production of IFNbeta by either untreated or MMC treated HTF. RESULTS: T-cell viability was significantly greater when T cells were cocultured with both untreated and growth-arrested HTF than when T cells were cultured alone (day 7, P = 0.0001). Neutralizing the action of IFNbeta blocked HTF-mediated T-cell rescue from apoptosis. Both untreated and growth arrested HTF secrete IFNbeta, and MMC at 0.4 mg/ml appeared to increase IFNbeta production. CONCLUSIONS: Cytokine deprivation-mediated T-cell apoptosis can be prevented by the action of IFNbeta secreted by both normal and growth-arrested HTF, which suggests that growth-arrested HTF can still participate in an aggressive wound-healing reaction by mediating a persistent inflammatory phase. This may partly explain why some trabeculectomies fail in high-risk patients, despite the use of antimetabolites. PMID- 11381058 TI - Immunogenicity of human amniotic membrane in experimental xenotransplantation. AB - PURPOSE: The immunogenic characterization of amniotic membrane is still unknown. This study was designed to examine the immunogenicity of human amniotic membrane, by using experimental xenotransplantation models. METHODS: Anti-human class I, class II, and Fas ligand monoclonal antibodies were used against cryopreserved amniotic membrane and cell viability tested for cryopreserved amniotic membrane. Amniotic membranes were then transplanted to the limbal area, intracorneal space, and under the kidney capsule. The scores of transparency and neovascularization after transplantation were recorded by slit lamp microscopy. Host cell infiltration was examined by hematoxylin-eosin or immunohistochemical staining. Control grafts were transplanted human cryopreserved skin grafts. RESULTS: Strong class I expression was observed in amniotic epithelium, mesenchymal cells, and fibroblasts in cryopreserved amniotic membrane. Some fibroblast cells unexpectedly expressed class II antigen. Fas ligand-positive cells were also detected in mesenchymal cells of amniotic stroma. Approximately 50% of epithelial cells of amniotic membrane cryopreserved for several months were still viable. In limbal transplantation, although some CD4(+) and CD8(+) T cells surrounded the amniotic graft, the response was mild. In intracorneal transplantation, all grafted amniotic membranes were accepted and clear, without host cell infiltration. In contrast, all skin grafts were rejected within 3 weeks after intracorneal transplantation. In amniotic membrane transplantation under the kidney capsule, extremely few host vessels and cells infiltrated the amniotic membrane; however, more host cells infiltrated the skin tissues under the kidney capsule. CONCLUSIONS: Amniotic membrane seems to be immune-privileged tissue and to contain some immunoregulatory factors, including HLA-G and Fas ligand. The amniotic membrane may be useful to supplement corneal collagen, and it may be applied not only to the ocular surface but also intracorneally. PMID- 11381059 TI - Expression of the chemokines MIP-1alpha, MCP-1, and RANTES in experimental autoimmune uveitis. AB - PURPOSE: To determine the location of the CC chemokines macrophage inflammatory protein (MIP)-1alpha, monocyte chemoattractant protein (MCP)-1, and regulated on activation of normal T-cell-expressed and secreted (RANTES) during disease progression in experimental autoimmune uveitis (EAU) and their relationship with the presence of the T helper cell (Th)1-type cytokine IFNgamma. METHODS: EAU was induced by immunization of Lewis rats with retinal extract. Consecutive cryostat sections were prepared from eyes at different stages of EAU, graded for severity of uveitis and stained by using antibodies to MCP-1, MIP-1alpha, and RANTES and to cell surface markers. Supernatants from superficial cervical lymph node cells were examined by ELISA for IFNgamma, IL-4, and IL-10. RESULTS: MIP-1alpha and IFNgamma were present most frequently and most extensively at peak disease but also were detectable in the choroid 8 days after immunization, before clinical disease onset. MCP-1 and RANTES were present at peak disease, but much less frequently. RANTES was occasionally found in the choroid before clinical disease. By days 19 to 21 after immunization, although infiltrating cells were present, there were only residual low levels of chemokine staining. MCP-1 and RANTES were detected on CD3-positive cells and on some ED1-positive cells, whereas MIP-1alpha was also associated with vessels and areas of exudate. Lymph node cells cultured from animals with peak disease had increased levels of IFNgamma and IL-10, but for IFNgamma this occurred only after stimulation in vitro with retinal extract. CONCLUSIONS: Although MCP-1 and RANTES were associated predominantly with cells infiltrating the retina, MIP-1alpha was also associated with resident cells. All three are likely to exacerbate EAU-MIP-1alpha, to the greatest degree. PMID- 11381060 TI - Inhibitory effects of antithrombin III against leukocyte rolling and infiltration during endotoxin-induced uveitis in rats. AB - PURPOSE: This study was designed to investigate the suppressive effects of antithrombin (AT)III on inflammatory reactions during endotoxin-induced uveitis (EIU) in rats by studying leukocyte-endothelium interactions. METHODS: EIU was induced in Lewis rats by footpad injection of lipopolysaccharide (LPS). ATIII was administered immediately after or at 6 hours after LPS injection. Its suppressive effects on inflammatory leukocyte behavior were evaluated in vivo with acridine orange digital fluorography. Clinical signs of inflammation were also examined, and aqueous humor (AH) was collected to evaluate leukocyte infiltration and protein leakage. In a separate experiment, P-selectin mRNA expression was studied in the iris-ciliary body (ICB) and the retina. RESULTS: After treatment with ATIII, leukocyte rolling was substantially inhibited along the retinal veins, suppressing subsequent leukocyte infiltration into the vitreous cavity. Similarly, leukocyte infiltration and protein leakage into the AH were significantly reduced with ATIII treatment. The clinical grade of EIU was substantially lower in ATIII-treated rats. In addition, delayed administration of ATIII after EIU induction significantly attenuated these inflammatory reactions. The levels of P-selectin mRNA expression in both ICB and retina, which were upregulated after LPS injection, were substantially lower in the ATIII-treated rats. CONCLUSIONS: ATIII treatment significantly inhibited inflammatory reactions induced with LPS. Its suppressive effects on P-selectin expression could contribute to the attenuation of leukocyte infiltration, possibly by inhibiting leukocyte rolling. The current findings suggest that ATIII may have a role in the management of patients with uveitis. PMID- 11381061 TI - Plasminogen Activators and Inhibitors in the Corneas of Mice Infected with Pseudomonas aeruginosa. AB - PURPOSE: To characterize the presence of plasminogen activators and their inhibitors in the corneas during the inflammatory response in naive and immunized mice intracorneally infected with Pseudomonas aeruginosa. METHODS: RT-PCR was used to detect gene expression for plasminogen activators and their inhibitors in naive and immunized mice. Immunoblot analysis, zymography, and ELISA were used to demonstrate the syntheses of these proteins. RESULTS: Naive mice intracorneally infected with P. aeruginosa showed a temporally enhanced expression of tissue type plasminogen activator (tPA), urokinase-type plasminogen activator (uPA), its receptor (uPAR), and plasminogen activator inhibitors 1 and 2 (PAI-1 and PAI-2), over a several-day holding period. Immunized mice demonstrated a lower and shorter expression of these factors over the same period. Expression of these factors at the mRNA and protein levels may have been due to enzymes and inhibitors present in inflammatory cells and in resident corneal cells. CONCLUSIONS: These results show a correlation between the overexpression of the PA system in infected naive mice as part of the inflammatory response, with eventual ocular destruction. Immunized mice exhibit a more balanced and shorter expression of the PA system, which may contribute to the restoration of corneal clarity. PMID- 11381062 TI - TNF receptor secretion after ex vivo adenoviral gene transfer to cornea and effect on in vivo graft survival. AB - PURPOSE: To explore the potential for adenovirus-mediated ex vivo gene transfer of a soluble tumor necrosis factor (TNF) receptor and evaluate the effect of transplanting the adenovirally transplanted corneas in vivo. METHODS: Rabbit corneal segments were transfected with replication-deficient adenovirus (AdTNFR) encoding a soluble TNF receptor fusion protein (TNFR-Ig). Production of TNFR-Ig was measured by using ELISA and bioassay. Corneas were transfected ex vivo with AdTNFR and then transplanted in vivo. Survival of AdTNFR-transfected corneas was compared with that of those treated either with a null vector control adenovirus (Ad0) or nontransfected control corneas. RESULTS: Ex vivo production of a molecule with TNF blocking bioactivity from AdTNFR-transfected corneas was demonstrated over a period of 4 weeks. Transplanted AdTNFR-transfected corneas showed a marginally increased survival time in vivo over nontransfected control corneas, but a significantly increased survival time over Ad0-treated control corneas. Ad0 treatment of corneal allografts before transplantation had a proinflammatory effect and accelerated the onset of corneal endothelial rejection. CONCLUSIONS: Adenoviral gene transfer is an effective means of transferring a gene encoding soluble TNFR-Ig to corneal endothelium, and ex vivo production of a biologically active secreted molecule was demonstrated for 4 weeks. However, in vivo, only a marginally increased survival was seen compared with control corneas. The introduction of this transgene using a less immunogenic vector may demonstrate prolongation of corneal allograft survival. PMID- 11381063 TI - Aey2, a new mutation in the betaB2-crystallin-encoding gene of the mouse. AB - PURPOSE: During an ethylnitrosourea (ENU) mutagenesis screen, mice were tested for the occurrence of dominant cataracts. One particular mutant was found that caused progressive opacity and was referred to as Aey2. The purpose of the study was to provide a morphologic description, to map the mutant gene, and to characterize the underlying molecular lesion. METHODS: Isolated lenses were photographed, and histologic sections of the eye were analyzed according to standard procedures. Linkage analysis was performed using a set of microsatellite markers covering all autosomal chromosomes. cDNA from candidate genes was amplified after reverse transcription of lens mRNA. RESULTS: The cortical opacification visible at eye opening progressed to an anterior suture cataract and reached its final phenotype as total opacity at 8 weeks of age. There was no obvious difference between heterozygous and homozygous mutants. The mutation was mapped to chromosome 5 proximal to the marker D5Mit138 (8.7 +/- 4.2 centimorgan [cM]) and distal to D5Mit15 (12.8 +/- 5.4 cM). No recombinations were observed to the markers D5Mit10 and D5Mit25. This position makes the genes within the betaA4/betaB-crystallin gene cluster excellent candidate genes. Sequence analysis revealed a mutation of T-->A at position 553 in the Crybb2 gene, leading to an exchange of Val for GLU: It affects the same region of the Crybb2 gene as in the Philly mouse. Correspondingly, the loss of the fourth Greek key motif is to be expected. CONCLUSIONS: The Aey2 mutant represents the second allele of Crybb2 in mice. Because an increasing number of beta- and gamma-crystallin mutations have been reported, a detailed phenotype-genotype correlation will allow a clearer functional understanding of beta- and gamma-crystallins. PMID- 11381064 TI - Evaluation of potentiating effect of a drop of lignocaine on tropicamide-induced mydriasis. AB - PURPOSE: To analyze whether preinstillation of lignocaine potentiates mydriasis by tropicamide in dark eyes and to determine possible mechanisms for this effect. METHODS: This investigation was conducted in two phases, the first being a double masked, placebo-controlled, randomized clinical trial, enrolling 60 healthy dark brown eyes in 30 subjects aged 7 to 58 years. The control eye received a drop of (nonlignocaine) placebo before tropicamide 1%, and the contralateral study eye received a 4% lignocaine drop 3-minutes before the 1 drop of tropicamide was administered. A ruled pupillometer recorded pupil diameters every 10 minutes for 50 minutes. In phase II, to elucidate pathomechanisms after lignocaine, corneal and tear parameters were compared with baseline records in a further 60 such eyes. RESULTS: Pupillary diameters in the study eyes increased by 3.62 +/- 0.75 mm, significantly more than in the placebo (control) group (P = 0.000). Ninety percent of study eyes attained the clinically significant 6-mm size with preinstillation of lignocaine-many more than the 67% of control eyes (P = 0.016). The median time to achieve this critical 6-mm size was significantly faster in the study group (P = 0.005). In phase II, the 1 drop 4% lignocaine did not show corneal changes with slit lamp or fluorescein staining and did not reduce media clarity or induce a significant change in tear pH. It markedly decreased Schirmer values (P = 0.000), reduced tear break-up time (P = 0.003), and increased corneal thickness measured by optical pachymetry (P = 0.010). CONCLUSIONS: The phase II findings indicate corneal microepithelial damage and reduced tearing. Both may enhance intraocular penetration and hence potentiation of tropicamide. This remarkable phenomenon could find use with many other important topical medications. PMID- 11381065 TI - Soluble TNF receptors in vitreoretinal proliferative disease. AB - PURPOSE: To measure vitreous levels of soluble TNF-receptors (sTNF-Rs) types I and II in eyes with rhegmatogenous retinal detachment (RRD), uncomplicated or complicated with proliferative vitreoretinopathy (PVR), and in eyes with proliferative diabetic retinopathy (PDR). To examine whether there is any relationship between vitreous levels of sTNF-Rs and clinical features of these conditions and between vitreous sTNF-Rs and TNFalpha levels and serum levels of sTNF-RS: METHODS: Vitreous levels of sTNF-Rs and TNFalpha were measured by enzyme linked immunosorbent assay in 30 eyes with PVR, 30 eyes with uncomplicated RRD, and 29 eyes with PDR. Vitreous from eyes of 10 deceased donors and 9 eyes with macular holes served as control specimens. Serum levels of sTNF-Rs were measured in 17 patients with PDR and 21 patients with PVR. RESULTS: Vitreous levels of sTNF-Rs I and II were increased in eyes with PVR, RRD, and PDR when compared with control eyes (P < 0.002). However, vitreous levels of sTNF-Rs I and II were higher in eyes with PVR than in eyes with RRD (P < 0.01) or PDR (P < 0.03). This contrasted with the findings that serum sTNF-Rs were higher in PDR than in PVR (P < 0.016) and that vitreous levels of TNFalpha were higher in eyes with PDR than in eyes with PVR (P < 0.0005). In PVR, vitreous sTNF-Rs levels were associated with the duration of retinal detachment, number of previous external operations, and grade of severity, whereas in PDR these levels were not related to the type or duration of diabetes or its complication with traction retinal detachment. CONCLUSIONS: These observations suggest the existence of TNF inhibitory mechanisms within the eye during retinal processes of inflammation and angiogenesis. That high vitreous levels of sTNF-Rs relate to severity of retinopathy suggests that these molecules may constitute reactive products of inflammation. Effective control of TNFalpha activity by sTNF-Rs within the retinal microenvironment may determine the outcome and severity of retinal proliferative conditions. PMID- 11381066 TI - The lipid composition of drusen, Bruch's membrane, and sclera by hot stage polarizing light microscopy. AB - PURPOSE: To detect and identify, in situ, the lipid composition of drusen, diffuse Bruch's membrane deposits, and sclera in aging human eyes using hot-stage polarizing microscopy (HSPM), a method that allows qualitative determination of lipid subtypes within histologic sections based on morphology and melting temperatures of liquid crystals as monitored by birefringence during heating and cooling. METHODS: Full-thickness buttons of the central macula and the periphery of human eyes from 17 patients were fixed in 5% calcium-buffered formalin. Frozen sections were stained with oil red O or Sudan black or were analyzed by HSPM. RESULTS: Birefringent anisotropic droplets ("maltese crosses") with melting characteristics of cholesterol esters were identified within diffuse Bruch's membrane deposits, drusen, and sclera. Deposits that melted from crystal to oil without any maltese cross formation when cooled were present in the sclera and are consistent with triglyceride-rich deposits. Deposits with optical properties consistent with phospholipids were identified in a single aged eye. Eyes from young donors did not show these changes. CONCLUSIONS: HSPM is a valuable technique for evaluating the nature of lipid deposits in aging eyes. Further studies are warranted to determine whether similar changes are also present in eyes with age-related macular degeneration. PMID- 11381067 TI - NADPH-diaphorase colocalization with somatostatin receptor subtypes sst2A and sst2B in the retina. AB - PURPOSE: To investigate the differential localization of somatotropin release inhibitory factor (SRIF) receptor subtypes (sst2A and sst2B) and their possible colocalization with reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase in the rat and rabbit retina. METHODS: Polyclonal antibodies raised against sst2A and sst2B receptors were applied to 10- to 14-microm cryostat sections of rat and rabbit retinas fixed in paraformaldehyde. NADPH-diaphorase reactivity was assessed histochemically. Double labeling was performed for sst2A or sst2B receptors with NADPH-diaphorase, and with markers for the cell types present in the retina (protein kinase C [PKC], tyrosine hydroxylase; [TH], calbindin, and recoverin). RESULTS: sst2A immunoreactivity was detected in rod bipolar cells and colocalized with NADPH-diaphorase in the rabbit, but not the rat, retina. sst2B was present only in photoreceptor cells of the rat and colocalized with NADPH-diaphorase. CONCLUSIONS: These results suggest that SRIF, acting through sst2A receptors in bipolar cells and sst2B receptors in photoreceptor cells, may affect nitric oxide function in the rabbit and rat retina. PMID- 11381068 TI - Novel CACNA1F mutations in Japanese patients with incomplete congenital stationary night blindness. AB - PURPOSE: Although it was reported that congenital stationary night blindness (CSNB) could be divided into complete and incomplete CSNB clinically in 1986, it was not until 1998 that the two types were found to be distinct clinical diseases by molecular genetic analysis. The purpose of this article is to report mutations in the retina-specific calcium channel alpha1-subunit gene (CACNA1F) in Japanese patients with incomplete CSNB and to describe the clinical features of these patients. METHODS: Seven patients from five separate Japanese families with incomplete CSNB were examined. Genomic DNA was extracted from leukocytes of the peripheral blood, and all 48 exons of the CACNA1F were amplified by polymerase chain reaction and directly sequenced. A complete ophthalmic examination was performed, including best corrected visual acuity, slit lamp and fundus examinations, fundus photography, and electroretinography. RESULTS: A mutation in the CACNA1F was identified in all the patients. The identified mutations were a missense mutation (Gly609Asp); a nonsense mutation (Arg913stop); a splice donor site mutation of G to C at nucleotide 2571+1; a G insertion at nucleotide 709, resulting in a frame shift with a predicted stop codon at codon 247; and a 4-bp deletion at nucleotides 271 to 274, with a replacement by an abnormal 34-bp sequence. Clinically, each patient had essentially normal fundi, mildly reduced corrected visual acuity, and slight myopia or hyperopia with astigmatism. Electrophysiologically, the mixed rod-cone ERG showed a negative configuration with recordable oscillatory potentials. The rod ERG was recordable but subnormal, and the cone and 30-Hz flicker ERGs were markedly depressed. CONCLUSIONS: Five novel mutations were identified in the CACNA1F in five Japanese families with incomplete CSNB, leading to the conclusion that in most Japanese patients, incomplete CSNB is caused by a CACNA1F mutation. PMID- 11381069 TI - Angiopoietin-1 upregulation by vascular endothelial growth factor in human retinal pigment epithelial cells. AB - PURPOSE: To determine whether vascular endothelial growth factor (VEGF) regulates angiopoietin (Ang)-1 and -2 expression in retinal pigment epithelial (RPE) cells. METHODS: Expression of VEGF, Ang1, and Ang2 in surgically removed human choroidal neovascular membranes (CNVMs) was analyzed by double-label confocal immunofluorescence microscopy. Total RNA was extracted from cultured human RPE cells treated with VEGF for mRNA analysis. Northern blot analysis was performed to examine the time course and dose response of Ang1 and Ang2 mRNA expression. mRNA stability and nuclear run-on analyses were performed. Secreted Ang1 and Ang2 protein levels in conditioned media from RPE cells were examined by Western blot analysis. RESULTS: Ang1 and Ang2 immunostaining colocalized with VEGF-positive stromal cells in human CNVMS: Ang1 and Ang2 mRNAs were expressed by cultured serum-starved RPE cells. VEGF upregulated Ang1 mRNA in a time- and dose-dependent manner without a significant change in Ang2 mRNA. Ang1 and Ang2 mRNAs in RPE cells were as stable as that of S18. VEGF stimulation further increased the half life of Ang1 mRNA, but did not alter its transcription rate. VEGF increased the amount of Ang1, but not Ang2, protein secreted into the medium. CONCLUSIONS: The colocalization of Ang1 and Ang2 with VEGF in CNVM stromal cells and the upregulation of Ang1 expression by VEGF in cultured RPE cells suggest that VEGF may selectively modulate Ang expression during CNV. PMID- 11381070 TI - Age and topographic variation of insulin-like growth factor-binding protein 2 in the human rpe. AB - PURPOSE: Previous studies have shown that insulin-like growth factor-binding protein (IGFBP)-2 is markedly upregulated in senescent RPE cells in vitro, and might therefore be a marker of senescent cells in vivo. This study was conducted to determine whether IGFBP-2 expression in human RPE cells from the macula and periphery varies with age in vivo. METHODS: Paraformaldehyde (4%)-fixed and optimal cutting temperature (OCT) compound-embedded human eyes from 17 patients were cryosectioned and subjected to high-sensitivity digoxigenin (DIG)-labeled cRNA in situ hybridization to determine the expression of IGFBP-2. Complementary immunohistochemistry experiments using a polyclonal anti-IGFBP-2 antibody were performed to confirm IGFBP-2 protein expression. Specimens were examined by light microscopy, and images were captured with a digital camera. The total numbers of RPE cells and IGFBP-2 mRNA expression-positive RPE cells were counted for each section, and the ratio of labeled RPE cells to total RPE cells counted was calculated for both macular and peripheral regions of each donor. RESULTS: IGFBP 2 mRNA expression was detected in the ganglion cell layer, inner and outer nuclear layers, and inner segments of photoreceptor cells in all 17 eyes. In 16 of 17 eyes, IGFBP-2 mRNA expression was detected in the RPE. In 11, the ratio of labeled cells to total RPE cells counted per section in the macula was 1.2 times greater than the ratio in the periphery (P = 0.008). The ratio of labeled RPE cells in the macula decreased with age (P = 0.0064). Immunohistochemistry studies for IGFBP-2 confirmed the expression pattern found by in situ hybridization. CONCLUSIONS: There is a topographical and age-related change in IGFBP-2 expression in RPE cells from human donor eyes. This distribution is likely not to represent senescent RPE cells in vivo. PMID- 11381071 TI - Differential chemokine regulation by Th2 cytokines during human RPE-monocyte coculture. AB - PURPOSE: To determine the effects of the potent anti-inflammatory Th2 cytokines, interleukin (IL)-4, -10, and -13, on IL-8 and monocyte chemoattractant protein (MCP) 1 production by human retinal pigment epithelial (HRPE) cells, monocytes, and HRPE cell-monocyte cocultures. METHODS: Enzyme-linked immunosorbent assays were performed to determine IL-8 and MCP-1 secretion by HRPE cells, monocytes, and HRPE cell-monocyte cocultures stimulated with IL-1beta or TNF-alpha, either alone, or in combination with IL-4, -10, or -13, at various time points. RESULTS: IL-4 and -13, but not IL-10, enhanced constitutive and TNF-alpha-induced HRPE IL 8 and MCP-1 secretion. IL-4 also enhanced IL-1beta-induced HRPE IL-8. IL-4 and 13 reduced monocyte IL-8 and MCP-1, whereas IL-10 reduced monocyte IL-8 but enhanced MCP-1. Overlay of monocytes onto HRPE cell cultures resulted in increased IL-8 and MCP-1 secretion. IL-8 secretion by HRPE cell-monocyte cocultures was inhibited by IL-4, -10, and -13, whereas MCP-1 was inhibited only by IL-10. These cytokines also inhibited IL-1beta potentiation of IL-8, but not MCP-1 secretion by cocultures. IL-4 enhanced TNF-alpha potentiation of chemokine secretion by cocultures, whereas IL-10 had no effects. IL-13 potentiated TNF alpha-induced MCP-1, but not IL-8 secretion by cocultures. CONCLUSIONS: IL-4, -10 and -13 have complex effects on chemokine secretion by HRPE cells, monocytes, and HRPE cell-monocyte cocultures. IL-10 appears to be the most consistently suppressive cytokine, suggesting potential therapeutic usefulness of IL-10 in the treatment of ocular inflammatory and proliferative diseases. PMID- 11381072 TI - Expression of prolactin gene and secretion of prolactin by rat retinal capillary endothelial cells. AB - PURPOSE: Prolactin fragments inhibit blood vessel formation, whereas anti prolactin antibodies induce angiogenesis in the cornea. Endothelial cells from brain capillaries and the umbilical vein produce prolactin, and this study was undertaken to determine whether retinal capillary endothelial cells could be a source for prolactin in the eye. METHODS: Primary cultures of rat retinal endothelial cells were investigated for the expression of prolactin mRNA by reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analysis and by in situ hybridization. The prolactin protein was analyzed by immunocytochemistry, enzyme-linked immunoabsorbent assay, Western blot analysis, and the Nb2-cell bioassay. The effect of prolactin and the 16-kDa prolactin fragment on retinal endothelial cell proliferation was investigated, and the expression of the cloned prolactin receptor was analyzed by RT-PCR and Southern blot analysis. RESULTS: Retinal endothelial cells expressed prolactin mRNA and full-length 23-kDa prolactin. Prolactin was observed in the cytoplasm of cells and in their conditioned medium at levels 300 times those described in endothelial cells from other vessels and species. Exogenous 16-kDa prolactin inhibited rat retinal endothelial cell proliferation, whereas 23-kDa prolactin was inactive. No evidence was obtained for the expression of the cloned prolactin receptor in these cells, but the prolactin receptor was amplified in whole rat retina. CONCLUSIONS: Endothelial cells from the microcirculation of rat retina produce and release prolactin. That the cloned prolactin receptor was not expressed in these cells argues against direct autocrine effects of prolactin. Possible paracrine effects are suggested by the expression of the prolactin receptor in retinal tissue. PMID- 11381073 TI - In vivo protection of photoreceptors from light damage by pigment epithelium derived factor. AB - PURPOSE: To determine whether pigment epithelium-derived factor (PEDF) exhibits neurotrophic and neuroprotective activities in vivo for photoreceptor cells. METHODS: Sprague-Dawley albino rats were injected intravitreally with 2 microg PEDF or a mixture of 1 microg basic fibroblast growth factor (bFGF)/1 microg PEDF in a volume of 1 microl phosphate-buffered saline (PBS). Animals were exposed to constant light for different periods at an illuminance level of 1200 to 1500 lux. The electroretinogram (ERG) waveforms of both eyes in the same animal were simultaneously recorded to evaluate functional protection. The morphologic protection was evaluated by quantitative histology. RESULTS: Intravitreal injection of PEDF before exposure to constant light resulted in significant morphologic and functional protection of photoreceptor cells in the retina of light-damaged rats. This protection depended on the duration and severity of light damage. The protection was eliminated by extending the light exposure to 10 days. Injection of PEDF at 0, 1, and 2 days after constant light exposure did not provide significant protection above that seen in PBS-injected eyes. Combination of PEDF with bFGF improved functional protection of photoreceptor cells. CONCLUSIONS: The data demonstrate that PEDF protected photoreceptor cells against light damage. This is significant, because it may open new avenues for the study of molecular mechanisms underlying degenerative processes. This, in turn, may lead to the development of therapeutic strategies for the prevention and treatment of degenerative diseases of the retina. PMID- 11381074 TI - Prevention of photoreceptor apoptosis by activation of the glucocorticoid receptor. AB - PURPOSE: Evidence has accumulated that excessive light exposure may promote age related and inherited retinal degeneration, in which photoreceptor death by apoptosis leads to loss of vision. In the current study, the effect of elevated corticosteroid levels on light-induced apoptosis of photoreceptors was determined. METHODS: Photoreceptor apoptosis was induced in retinas of BALB/c mice by exposure to diffuse white light. High levels of corticosteroids were induced, either endogenously (fasting-mediated stress) or by a single intraperitoneal injection of dexamethasone (DEX). Photoreceptor damage was assessed morphologically and by electroretinography. Glucocorticoid receptor (GR) and activator protein (AP)-1 activities were shown by Western blot analysis and electrophoretic mobility shift assay (EMSA) of retinal nuclear extracts. RESULTS: Fasting and injection of DEX led to an activation of GR in the retina, as judged by its translocation to the nucleus of retinal cells. On induction of GR activity before light exposure, AP-1 activity, normally induced by damaging doses of light, remained at basal levels. Both treatments completely prevented photoreceptor apoptosis and preserved retinal function. CONCLUSIONS: Activity of the transcription factor AP-1 is associated with light-induced apoptosis. In the current study, pharmacologic suppression of AP-1 activity protected against light damage. Inhibition of AP-1 activity may have occurred by the protein-protein interaction of GR and AP-1. PMID- 11381075 TI - Signaling pathways for glycated human serum albumin-induced IL-8 and MCP-1 secretion in human RPE cells. AB - PURPOSE: To determine the signal mediators involved in glycated human serum albumin (GHSA) stimulation of interleukin (IL)-8 and monocyte chemotactic protein (MCP)-1 secretion in human retinal pigment epithelium (hRPE) cells. METHODS: hRPE cells were stimulated by GHSA in the presence or absence of a series of kinase inhibitors. The induced IL-8 and MCP-1 mRNA and proteins were determined by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA). Western blot analysis, electrophoretic mobility shift assay, and immunohistochemical staining were used to analyze activation of signaling mediators and transcription factors. RESULTS: Incubation of hRPE cells with GHSA resulted in rapid activation of Raf-1, extracellular signal-regulated protein kinases (ERK) 1/2, p38, and the transcription factor nuclear factor (NF) kappaB. Coincubation of hRPE cells with the mitogen-activated protein (MAP) kinase (MEK) inhibitor U0126; NF-kappaB inhibitors BAY11-7085, caffeic acid phenethyl ester (CAPE), parthenolide, and curcumin; protein kinase (PK)C inhibitor Ro318220; and protein tyrosine kinase (PTK) inhibitor genistein largely eliminated most of the stimulated production of IL-8 and MCP-1. Combined inhibition of MEK by U0126, p38 by SB202190, and Janus kinase (jak) by AG490 revealed that GHSA stimulation of IL-8 production was predominately mediated by MEK and to a lesser extent by p38 pathways, whereas activation of MEK, p38, and jak was required for maximal MCP-1 induction. Moreover, GHSA-stimulated IL-8 secretion was more sensitive to U0126 (50% inhibitory concentration [IC(50)] = 0.5 microM) than MCP-1 (IC(50) = 10 microM). CONCLUSIONS: GHSA stimulates hRPE IL 8 and MCP-1 production through divergent and overlapping, but not identical, intracellular signaling cascades. GHSA induces activation of a series of kinases including PKC, PTK, MAPK, p38, and jak and the transcription factor NF-kappaB. The Raf/MAPK pathway plays an essential role in GHSA signaling. PMID- 11381076 TI - Retinal transplants restore visually evoked responses in rats with photoreceptor degeneration. AB - PURPOSE: To assess whether transplantation of intact sheets of fetal retina with retinal pigment epithelium (RPE) into a retina with photoreceptor degeneration restores visually evoked responses. METHODS: Sheets of fetal retina with RPE were transplanted into the subretinal space of Royal College of Surgeons (RCS) rats at 37 to 69 days of age. Sixty-three days to 10 months after transplantation, multiunit visual responses were recorded in the superior colliculus (SC) of transplanted rats, age-matched untransplanted rats, and rats with sham surgery. RESULTS: In 19 of 29 RCS rats with transplants, visually evoked responses were recorded from and restricted to a small area of the SC that corresponds topographically to the portion of the retina in which the transplant was placed. Outside of this area, no visual responses were evoked. Visually evoked responses were never recorded in age-matched, nontransplanted RCS rats. Visually evoked responses were recorded in 6 of 13 RCS rats with sham surgery, but these responses were significantly different from responses in rats with transplants. CONCLUSIONS: These results demonstrate that this transplantation technique restores visually evoked responses in the brain. Although the underlying mechanism is unknown, we propose that the central visual response results from increased synaptic efficacy within the host retina. If it can be established that functional connections between the transplant and the host retina produce the effect, then it would indicate that the technique could be explored as a therapeutic strategy in some diseases of retinal degeneration. PMID- 11381077 TI - Membrane-type 1 matrix metalloproteinase cleaves CD44 and promotes cell migration. AB - Migratory cells including invasive tumor cells frequently express CD44, a major receptor for hyaluronan and membrane-type 1 matrix metalloproteinase (MT1-MMP) that degrades extracellular matrix at the pericellular region. In this study, we demonstrate that MT1-MMP acts as a processing enzyme for CD44H, releasing it into the medium as a soluble 70-kD fragment. Furthermore, this processing event stimulates cell motility; however, expression of either CD44H or MT1-MMP alone did not stimulate cell motility. Coexpression of MT1-MMP and mutant CD44H lacking the MT1-MMP-processing site did not result in shedding and did not promote cell migration, suggesting that the processing of CD44H by MT1-MMP is critical in the migratory stimulation. Moreover, expression of the mutant CD44H inhibited the cell migration promoted by CD44H and MT1-MMP in a dominant-negative manner. The pancreatic tumor cell line, MIA PaCa-2, was found to shed the 70-kD CD44H fragment in a MT1-MMP-dependent manner. Expression of the mutant CD44H in the cells as well as MMP inhibitor treatment effectively inhibited the migration, suggesting that MIA PaCa-2 cells indeed use the CD44H and MT1-MMP as migratory devices. These findings revealed a novel interaction of the two molecules that have each been implicated in tumor cell migration and invasion. PMID- 11381078 TI - Inhibition of beta(2) integrin-mediated leukocyte cell adhesion by leucine leucine-glycine motif-containing peptides. AB - Many integrins mediate cell attachment to the extracellular matrix by recognizing short tripeptide sequences such as arginine-glycine-aspartic acid and leucine aspartate-valine. Using phage display, we have now found that the leukocyte specific beta(2) integrins bind sequences containing a leucine-leucine-glycine (LLG) tripeptide motif. An LLG motif is present on intercellular adhesion molecule (ICAM)-1, the major beta(2) integrin ligand, but also on several matrix proteins, including von Willebrand factor. We developed a novel beta(2) integrin antagonist peptide CPCFLLGCC (called LLG-C4), the structure of which was determined by nuclear magnetic resonance. The LLG-C4 peptide inhibited leukocyte adhesion to ICAM-1, and, interestingly, also to von Willebrand factor. When immobilized on plastic, the LLG-C4 sequence supported the beta(2) integrin mediated leukocyte adhesion, but not beta(1) or beta(3) integrin-mediated cell adhesion. These results suggest that LLG sequences exposed on ICAM-1 and on von Willebrand factor at sites of vascular injury play a role in the binding of leukocytes, and LLG-C4 and peptidomimetics derived from it could provide a therapeutic approach to inflammatory reactions. PMID- 11381079 TI - Cyclin-dependent kinase inhibitor p27(Kip1) is required for mouse mammary gland morphogenesis and function. AB - We have studied the role of the cyclin-dependent kinase (Cdk) inhibitor p27(Kip1) in postnatal mammary gland morphogenesis. Based on its ability to negatively regulate cyclin/Cdk function, loss of p27 may result in unrestrained cellular proliferation. However, recent evidence about the stabilizing effect of p27 on cyclin D1-Cdk4 complexes suggests that p27 deficiency might recapitulate the hypoplastic mammary phenotype of cyclin D1-deficient animals. These hypotheses were investigated in postnatal p27-deficient (p27(-/-)), hemizygous (p27(+/)-), or wild-type (p27(+/+)) mammary glands. Mammary glands from p27(+/)- mice displayed increased ductal branching and proliferation with delayed postlactational involution. In contrast, p27(-/-) mammary glands or wild-type mammary fat pads reconstituted with p27(-/-) epithelium produced the opposite phenotype: hypoplasia, low proliferation, decreased ductal branching, impaired lobuloalveolar differentiation, and inability to lactate. The association of cyclin D1 with Cdk4, the kinase activity of Cdk4 against pRb in vitro, the nuclear localization of cyclin D1, and the stability of cyclin D1 were all severely impaired in p27(-/-) mammary epithelial cells compared with p27(+/+) and p27(+/-) mammary epithelial cells. Therefore, p27 is required for mammary gland development in a dose-dependent fashion and positively regulates cyclin D-Cdk4 function in the mammary gland. PMID- 11381080 TI - Endothelial cell laminin isoforms, laminins 8 and 10, play decisive roles in T cell recruitment across the blood-brain barrier in experimental autoimmune encephalomyelitis. AB - An active involvement of blood-brain barrier endothelial cell basement membranes in development of inflammatory lesions in the central nervous system (CNS) has not been considered to date. Here we investigated the molecular composition and possible function of the extracellular matrix encountered by extravasating T lymphocytes during experimental autoimmune encephalomyelitis (EAE). Endothelial basement membranes contained laminin 8 (alpha4beta1gamma1) and/or 10 (alpha5beta1gamma1) and their expression was influenced by proinflammatory cytokines or angiostatic agents. T cells emigrating into the CNS during EAE encountered two biochemically distinct basement membranes, the endothelial (containing laminins 8 and 10) and the parenchymal (containing laminins 1 and 2) basement membranes. However, inflammatory cuffs occurred exclusively around endothelial basement membranes containing laminin 8, whereas in the presence of laminin 10 no infiltration was detectable. In vitro assays using encephalitogenic T cell lines revealed adhesion to laminins 8 and 10, whereas binding to laminins 1 and 2 could not be induced. Downregulation of integrin alpha6 on cerebral endothelium at sites of T cell infiltration, plus a high turnover of laminin 8 at these sites, suggested two possible roles for laminin 8 in the endothelial basement membrane: one at the level of the endothelial cells resulting in reduced adhesion and, thereby, increased penetrability of the monolayer; and secondly at the level of the T cells providing direct signals to the transmigrating cells. PMID- 11381081 TI - An atomic model of actin filaments cross-linked by fimbrin and its implications for bundle assembly and function. AB - Actin bundles have profound effects on cellular shape, division, adhesion, motility, and signaling. Fimbrin belongs to a large family of actin-bundling proteins and is involved in the formation of tightly ordered cross-linked bundles in the brush border microvilli and in the stereocilia of inner ear hair cells. Polymorphism in these three-dimensional (3D) bundles has prevented the detailed structural characterization required for in-depth understanding of their morphogenesis and function. Here, we describe the structural characterization of two-dimensional arrays of actin cross-linked with human T-fimbrin. Structural information obtained by electron microscopy, x-ray crystallography, and homology modeling allowed us to build the first molecular model for the complete actin fimbrin cross-link. The restriction of the arrays to two dimensions allowed us to deduce the spatial relationship between the components, the mode of fimbrin cross linking, and the flexibility within the cross-link. The atomic model of the fimbrin cross-link, the cross-linking rules deduced from the arrays, and the hexagonal packing of actin bundles in situ were all combined to generate an atomic model for 3D actin-fimbrin bundles. Furthermore, the assembly of the actin fimbrin arrays suggests coupling between actin polymerization, fimbrin binding, and crossbridge formation, presumably achieved by a feedback between conformational changes and changes in affinity. PMID- 11381082 TI - COOH-terminal truncations promote proteasome-dependent degradation of mature cystic fibrosis transmembrane conductance regulator from post-Golgi compartments. AB - Impaired biosynthetic processing of the cystic fibrosis (CF) transmembrane conductance regulator (CFTR), a cAMP-regulated chloride channel, constitutes the most common cause of CF. Recently, we have identified a distinct category of mutation, caused by premature stop codons and frameshift mutations, which manifests in diminished expression of COOH-terminally truncated CFTR at the cell surface. Although the biosynthetic processing and plasma membrane targeting of truncated CFTRs are preserved, the turnover of the complex-glycosylated mutant is sixfold faster than its wild-type (wt) counterpart. Destabilization of the truncated CFTR coincides with its enhanced susceptibility to proteasome-dependent degradation from post-Golgi compartments globally, and the plasma membrane specifically, determined by pulse-chase analysis in conjunction with cell surface biotinylation. Proteolytic cleavage of the full-length complex-glycosylated wt and degradation intermediates derived from both T70 and wt CFTR requires endolysosomal proteases. The enhanced protease sensitivity in vitro and the decreased thermostability of the complex-glycosylated T70 CFTR in vivo suggest that structural destabilization may account for the increased proteasome susceptibility and the short residence time at the cell surface. These in turn are responsible, at least in part, for the phenotypic manifestation of CF. We propose that the proteasome-ubiquitin pathway may be involved in the peripheral quality control of other, partially unfolded membrane proteins as well. PMID- 11381083 TI - The orphan nuclear estrogen receptor-related receptor alpha (ERRalpha) is expressed throughout osteoblast differentiation and regulates bone formation in vitro. AB - The orphan nuclear estrogen receptor-related receptor alpha (ERRalpha), is expressed by many cell types, but is very highly expressed by osteoblastic cells in which it transactivates at least one osteoblast-associated gene, osteopontin. To study the putative involvement of ERRalpha in bone, we first assessed its expression in rat calvaria (RC) in vivo and in RC cells in vitro. ERRalpha mRNA and protein were expressed at all developmental stages from early osteoprogenitors to bone-forming osteoblasts, but protein was most abundant in mature cuboidal osteoblasts. To assess a functional role for ERRalpha in osteoblast differentiation and bone formation, we blocked its expression by antisense oligonucleotides in either proliferating or differentiating RC cell cultures and found inhibition of cell growth and a proliferation-independent inhibition of differentiation. On the other hand, ERRalpha overexpression in RC cells increased differentiation and maturation of progenitors to mature bone forming cells. Our findings show that ERRalpha is highly expressed throughout the osteoblast developmental sequence and plays a physiological role in differentiation and bone formation at both proliferation and differentiation stages. In addition, we found that manipulation of receptor levels in the absence of known ligand is a fruitful approach for functional analysis of this orphan receptor and identification of potential target genes. PMID- 11381084 TI - The small muscle-specific protein Csl modifies cell shape and promotes myocyte fusion in an insulin-like growth factor 1-dependent manner. AB - We have isolated a murine cDNA encoding a 9-kD protein, Chisel (Csl), in a screen for transcriptional targets of the cardiac homeodomain factor Nkx2-5. Csl transcripts were detected in atria and ventricles of the heart and in all skeletal muscles and smooth muscles of the stomach and pulmonary veins. Csl protein was distributed throughout the cytoplasm in fetal muscles, although costameric and M-line localization to the muscle cytoskeleton became obvious after further maturation. Targeted disruption of Csl showed no overt muscle phenotype. However, ectopic expression in C2C12 myoblasts induced formation of lamellipodia in which Csl protein became tethered to membrane ruffles. Migration of these cells was retarded in a monolayer wound repair assay. Csl-expressing myoblasts differentiated and fused normally, although in the presence of insulin like growth factor (IGF)-1 they showed dramatically enhanced fusion, leading to formation of large dysmorphogenic "myosacs." The activities of transcription factors nuclear factor of activated T cells (NFAT) and myocyte enhancer-binding factor (MEF)2, were also enhanced in an IGF-1 signaling-dependent manner. The dynamic cytoskeletal localization of Csl and its dominant effects on cell shape and behavior and transcription factor activity suggest that Csl plays a role in the regulatory network through which muscle cells coordinate their structural and functional states during growth, adaptation, and repair. PMID- 11381085 TI - Cathepsin B acts as a dominant execution protease in tumor cell apoptosis induced by tumor necrosis factor. AB - Death receptors can trigger cell demise dependent or independent of caspases. In WEHI-S fibrosarcoma cells, tumor necrosis factor (TNF) induced an increase in cytosolic cathepsin B activity followed by death with apoptotic features. Surprisingly, this process was enhanced by low, but effectively inhibiting, concentrations of pan-caspase inhibitors. Contrary to caspase inhibitors, a panel of pharmacological cathepsin B inhibitors, the endogenous cathepsin inhibitor cystatin A as well as antisense-mediated depletion of cathepsin B rescued WEHI-S cells from apoptosis triggered by TNF or TNF-related apoptosis-inducing ligand. Thus, cathepsin B can take over the role of the dominant execution protease in death receptor-induced apoptosis. The conservation of this alternative execution pathway was further examined in other tumor cell lines. Here, cathepsin B acted as an essential downstream mediator of TNF-triggered and caspase-initiated apoptosis cascade, whereas apoptosis of primary cells was only minimally dependent on cathepsin B. These data imply that cathepsin B, which is commonly overexpressed in human primary tumors, may have two opposing roles in malignancy, reducing it by its proapoptotic features and enhancing it by its known facilitation of invasion. PMID- 11381086 TI - Feedback inhibition of the unfolded protein response by GADD34-mediated dephosphorylation of eIF2alpha. AB - Phosphorylation of the alpha subunit of eukaryotic translation initiation factor 2 (eIF2alpha) on serine 51 integrates general translation repression with activation of stress-inducible genes such as ATF4, CHOP, and BiP in the unfolded protein response. We sought to identify new genes active in this phospho eIF2alpha-dependent signaling pathway by screening a library of recombinant retroviruses for clones that inhibit the expression of a CHOP::GFP reporter. A retrovirus encoding the COOH terminus of growth arrest and DNA damage gene (GADD)34, also known as MYD116 (Fornace, A.J., D.W. Neibert, M.C. Hollander, J.D. Luethy, M. Papathanasiou, J. Fragoli, and N.J. Holbrook. 1989. Mol. Cell. Biol. 9:4196-4203; Lord K.A., B. Hoffman-Lieberman, and D.A. Lieberman. 1990. Nucleic Acid Res. 18:2823), was isolated and found to attenuate CHOP (also known as GADD153) activation by both protein malfolding in the endoplasmic reticulum, and amino acid deprivation. Despite normal activity of the cognate stress-inducible eIF2alpha kinases PERK (also known as PEK) and GCN2, phospho-eIF2alpha levels were markedly diminished in GADD34-overexpressing cells. GADD34 formed a complex with the catalytic subunit of protein phosphatase 1 (PP1c) that specifically promoted the dephosphorylation of eIF2alpha in vitro. Mutations that interfered with the interaction with PP1c prevented the dephosphorylation of eIF2alpha and blocked attenuation of CHOP by GADD34. Expression of GADD34 is stress dependent, and was absent in PERK(-)/- and GCN2(-)/- cells. These findings implicate GADD34 mediated dephosphorylation of eIF2alpha in a negative feedback loop that inhibits stress-induced gene expression, and that might promote recovery from translational inhibition in the unfolded protein response. PMID- 11381087 TI - Activation of the Met receptor by cell attachment induces and sustains hepatocellular carcinomas in transgenic mice. AB - Overexpression is the most common abnormality of receptor tyrosine kinases (RTKs) in human tumors. It is presumed that overexpression leads to constitutive activation of RTKs, but the mechanism of that activation has been uncertain. Here we show that overexpression of the Met RTK allows activation of the receptor by cell attachment and that this form of activation can be tumorigenic. Transgenic mice that overexpressed Met in hepatocytes developed hepatocellular carcinoma (HCC), one of the human tumors in which Met has been implicated previously. The tumorigenic Met was activated by cell attachment rather than by ligand. Inactivation of the transgene led to regression of even highly advanced tumors, apparently mediated by apoptosis and cessation of cellular proliferation. These results reveal a previously unappreciated mechanism by which the tumorigenic action of RTKs can be mediated, provide evidence that Met may play a role in both the genesis and maintenance of HCC, and suggest that Met may be a beneficial therapeutic target in tumors that overexpress the receptor. PMID- 11381088 TI - Local photorelease of caged thymosin beta4 in locomoting keratocytes causes cell turning. AB - The broad aim of this work was to explore the feasibility of using light-directed perturbation techniques to study cell locomotion. Specifically, a caged form of thymosin beta4 (Tbeta4) was photoactivated in a defined local region of locomoting fish scale keratocytes and the resulting perturbation of locomotion was studied. Purified Tbeta4 was produced in an inactive form by "caging" with ([n-nitroveratryl]oxy)chlorocarbamate. In vitro spectrophotofluorometric assays indicated that caged Tbeta4 did not change the normal actin polymerization kinetics, whereas photoactivated Tbeta4 significantly inhibited actin polymerization. With an a priori knowledge of the cytoplasmic diffusion coefficient of Tbeta4 as measured by fluorescence recovery after photobleaching experiments, the rapid sequestration of actin monomers by uncaged Tbeta4 and the consequent reduction in the diffusional spread of the Tbeta4-actin complex were predicted using Virtual Cell software (developed at the Center for Biomedical Imaging Technology, University of Connecticut Health Center). These simulations demonstrated that locally photoactivating Tbeta4 in keratocytes could potentially elicit a regional locomotory response. Indeed, when caged Tbeta4 was locally photoactivated at the wings of locomoting keratocytes, specific turning about the irradiated region was observed, whereas various controls were negative. Additionally, loading of exogenous Tbeta4 into both keratocytes and fibroblasts caused very rapid disassembly of actin filaments and reduction of cellular contractility. Based on these results, a mechanical model is proposed for the turning behavior of keratocytes in response to photoreleased Tbeta4. PMID- 11381089 TI - E-cadherin suppresses cellular transformation by inhibiting beta-catenin signaling in an adhesion-independent manner. AB - E-cadherin is a tumor suppressor protein with a well-established role in cell cell adhesion. Adhesion could contribute to tumor suppression either by physically joining cells or by facilitating other juxtacrine signaling events. Alternatively, E-cadherin tumor suppressor activity could result from binding and antagonizing the nuclear signaling function of beta-catenin, a known proto oncogene. To distinguish between an adhesion- versus a beta-catenin signaling dependent mechanism, chimeric cadherin constructs were expressed in the SW480 colorectal tumor cell line. Expression of wild-type E-cadherin significantly inhibits the growth of this cell line. Growth inhibitory activity is retained by all constructs that have the beta-catenin binding region of the cytoplasmic domain but not by E-cadherin constructs that exhibit adhesive activity, but lack the beta-catenin binding region. This growth suppression correlates with a reduction in beta-catenin/T cell factor (TCF) reporter gene activity. Importantly, direct inhibition of beta-catenin/TCF signaling inhibits the growth of SW480 cells, and the growth inhibitory activity of E-cadherin is rescued by constitutively activated forms of TCF. Thus, the growth suppressor activity of E cadherin is adhesion independent and results from an inhibition of the beta catenin/TCF signaling pathway, suggesting that loss of E-cadherin expression can contribute to upregulation of this pathway in human cancers. E-cadherin-mediated growth suppression was not accompanied by overall depletion of beta-catenin from the cytosol and nucleus. This appears to be due to the existence of a large pool of cytosolic beta-catenin in SW480 cells that is refractory to both cadherin binding and TCF binding. Thus, a small pool of beta-catenin that can bind TCF (i.e., the transcriptionally active pool) can be selectively depleted by E cadherin expression. The existence of functionally distinct pools of cytosolic beta-catenin suggests that there are mechanisms to regulate beta-catenin signaling in addition to controlling its level of accumulation. PMID- 11381090 TI - Molecular chaperones in the yeast endoplasmic reticulum maintain the solubility of proteins for retrotranslocation and degradation. AB - Endoplasmic reticulum (ER)-associated degradation (ERAD) is the process by which aberrant proteins in the ER lumen are exported back to the cytosol and degraded by the proteasome. Although ER molecular chaperones are required for ERAD, their specific role(s) in this process have been ill defined. To understand how one group of interacting lumenal chaperones facilitates ERAD, the fates of pro-alpha factor and a mutant form of carboxypeptidase Y were examined both in vivo and in vitro. We found that these ERAD substrates are stabilized and aggregate in the ER at elevated temperatures when BiP, the lumenal Hsp70 molecular chaperone, is mutated, or when the genes encoding the J domain-containing proteins Jem1p and Scj1p are deleted. In contrast, deletion of JEM1 and SCJ1 had little effect on the ERAD of a membrane protein. These results suggest that one role of the BiP, Jem1p, and Scj1p chaperones is to maintain lumenal ERAD substrates in a retrotranslocation-competent state. PMID- 11381091 TI - Growth cone collapse through coincident loss of actin bundles and leading edge actin without actin depolymerization. AB - Repulsive guidance cues can either collapse the whole growth cone to arrest neurite outgrowth or cause asymmetric collapse leading to growth cone turning. How signals from repulsive cues are translated by growth cones into this morphological change through rearranging the cytoskeleton is unclear. We examined three factors that are able to induce the collapse of extending Helisoma growth cones in conditioned medium, including serotonin, myosin light chain kinase inhibitor, and phorbol ester. To study the cytoskeletal events contributing to collapse, we cultured Helisoma growth cones on polylysine in which lamellipodial collapse was prevented by substrate adhesion. We found that all three factors that induced collapse of extending growth cones also caused actin bundle loss in polylysine-attached growth cones without loss of actin meshwork. In addition, actin bundle loss correlated with specific filamentous actin redistribution away from the leading edge that is characteristic of repulsive factors. Finally, we provide direct evidence using time-lapse studies of extending growth cones that actin bundle loss paralleled collapse. Taken together, these results suggest that actin bundles could be a common cytoskeletal target of various collapsing factors, which may use different signaling pathways that converge to induce growth cone collapse. PMID- 11381092 TI - Mba1, a novel component of the mitochondrial protein export machinery of the yeast Saccharomyces cerevisiae. AB - The biogenesis of mitochondria requires the integration of many proteins into the inner membrane from the matrix side. The inner membrane protein Oxa1 plays an important role in this process. We identified Mba1 as a second mitochondrial component that is required for efficient protein insertion. Like Oxa1, Mba1 specifically interacts both with mitochondrial translation products and with conservatively sorted, nuclear-encoded proteins during their integration into the inner membrane. Oxa1 and Mba1 overlap in function and substrate specificity, but both can act independently of each other. We conclude that Mba1 is part of the mitochondrial protein export machinery and represents the first component of a novel Oxa1-independent insertion pathway into the mitochondrial inner membrane. PMID- 11381093 TI - Nucleolar assembly of the rRNA processing machinery in living cells. AB - To understand how nuclear machineries are targeted to accurate locations during nuclear assembly, we investigated the pathway of the ribosomal RNA (rRNA) processing machinery towards ribosomal genes (nucleolar organizer regions [NORs]) at exit of mitosis. To follow in living cells two permanently transfected green fluorescence protein-tagged nucleolar proteins, fibrillarin and Nop52, from metaphase to G1, 4-D time-lapse microscopy was used. In early telophase, fibrillarin is concentrated simultaneously in prenucleolar bodies (PNBs) and NORs, whereas PNB-containing Nop52 forms later. These distinct PNBs assemble at the chromosome surface. Analysis of PNB movement does not reveal the migration of PNBs towards the nucleolus, but rather a directional flow between PNBs and between PNBs and the nucleolus, ensuring progressive delivery of proteins into nucleoli. This delivery appeared organized in morphologically distinct structures visible by electron microscopy, suggesting transfer of large complexes. We propose that the temporal order of PNB assembly and disassembly controls nucleolar delivery of these proteins, and that accumulation of processing complexes in the nucleolus is driven by pre-rRNA concentration. Initial nucleolar formation around competent NORs appears to be followed by regroupment of the NORs into a single nucleolus 1 h later to complete the nucleolar assembly. This demonstrates the formation of one functional domain by cooperative interactions between different chromosome territories. PMID- 11381094 TI - Stonin 2: an adaptor-like protein that interacts with components of the endocytic machinery. AB - Endocytosis of cell surface proteins is mediated by a complex molecular machinery that assembles on the inner surface of the plasma membrane. Here, we report the identification of two ubiquitously expressed human proteins, stonin 1 and stonin 2, related to components of the endocytic machinery. The human stonins are homologous to the Drosophila melanogaster stoned B protein and exhibit a modular structure consisting of an NH(2)-terminal proline-rich domain, a central region of homology specific to the stonins, and a COOH-terminal region homologous to the mu subunits of adaptor protein (AP) complexes. Stonin 2, but not stonin 1, interacts with the endocytic machinery proteins Eps15, Eps15R, and intersectin 1. These interactions occur via two NPF motifs in the proline-rich domain of stonin 2 and Eps15 homology domains of Eps15, Eps15R, and intersectin 1. Stonin 2 also interacts indirectly with the adaptor protein complex, AP-2. In addition, stonin 2 binds to the C2B domains of synaptotagmins I and II. Overexpression of GFP stonin 2 interferes with recruitment of AP-2 to the plasma membrane and impairs internalization of the transferrin, epidermal growth factor, and low density lipoprotein receptors. These observations suggest that stonin 2 is a novel component of the general endocytic machinery. PMID- 11381095 TI - The yeast class V myosins, Myo2p and Myo4p, are nonprocessive actin-based motors. AB - The motor properties of the two yeast class V myosins, Myo2p and Myo4p, were examined using in vitro motility assays. Both myosins are active motors with maximum velocities of 4.5 microm/s for Myo2p and 1.1 microm/s for Myo4p. Myo2p motility is Ca(2+) insensitive. Both myosins have properties of a nonprocessive motor, unlike chick myosin-Va (M5a), which behaves as a processive motor when assayed under identical conditions. Additional support for the idea that Myo2p is a nonprocessive motor comes from actin cosedimentation assays, which show that Myo2p has a low affinity for F-actin in the presence of ATP and Ca(2+), unlike chick brain M5a. These studies suggest that if Myo2p functions in organelle transport, at least five molecules of Myo2p must be present per organelle to promote directed movement. PMID- 11381096 TI - MAGI-1c: a synaptic MAGUK interacting with muSK at the vertebrate neuromuscular junction. AB - The muscle-specific receptor tyrosine kinase (MuSK) forms part of a receptor complex, activated by nerve-derived agrin, that orchestrates the differentiation of the neuromuscular junction (NMJ). The molecular events linking MuSK activation with postsynaptic differentiation are not fully understood. In an attempt to identify partners and/or effectors of MuSK, cross-linking and immunopurification experiments were performed in purified postsynaptic membranes from the Torpedo electrocyte, a model system for the NMJ. Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) analysis was conducted on both cross-link products, and on the major peptide coimmunopurified with MuSK; this analysis identified a polypeptide corresponding to the COOH-terminal fragment of membrane associated guanylate kinase (MAGUK) with inverted domain organization (MAGI)-1c. A bona fide MAGI-1c (150 kD) was detected by Western blotting in the postsynaptic membrane of Torpedo electrocytes, and in a high molecular mass cross-link product of MuSK. Immunofluorescence experiments showed that MAGI-1c is localized specifically at the adult rat NMJ, but is absent from agrin-induced acetylcholine receptor clusters in myotubes in vitro. In the central nervous system, MAGUKs play a primary role as scaffolding proteins that organize cytoskeletal signaling complexes at excitatory synapses. Our data suggest that a protein from the MAGUK family is involved in the MuSK signaling pathway at the vertebrate NMJ. PMID- 11381097 TI - Circulating skeletal stem cells. AB - We report the isolation of adherent, clonogenic, fibroblast-like cells with osteogenic and adipogenic potential from the blood of four mammalian species. These cells phenotypically resemble but are distinguishable from skeletal stem cells found in bone marrow (stromal stem cells, "mesenchymal stem cells"). The osteogenic potential of the blood-borne cells was proven by an in vivo transplantation assay in which either polyclonal or single colony-derived strains were transplanted into the subcutis of immunocompromised mice, and the donor origin of the fully differentiated bone cells was proven using species-specific probes. This is the first definitive proof of the existence of circulating skeletal stem cells in mammals. PMID- 11381098 TI - PDZ protein interactions regulating glutamate receptor function and plasticity. PMID- 11381099 TI - Pneumococcal virulence factors: structure and function. AB - The overall goal for this review is to summarize the current body of knowledge about the structure and function of major known antigens of Streptococcus pneumoniae, a major gram-positive bacterial pathogen of humans. This information is then related to the role of these proteins in pneumococcal pathogenesis and in the development of new vaccines and/or other antimicrobial agents. S. pneumoniae is the most common cause of fatal community-acquired pneumonia in the elderly and is also one of the most common causes of middle ear infections and meningitis in children. The present vaccine for the pneumococcus consists of a mixture of 23 different capsular polysaccharides. While this vaccine is very effective in young adults, who are normally at low risk of serious disease, it is only about 60% effective in the elderly. In children younger than 2 years the vaccine is ineffective and is not recommended due to the inability of this age group to mount an antibody response to the pneumococcal polysaccharides. Antimicrobial drugs such as penicillin have diminished the risk from pneumococcal disease. Several pneumococcal proteins including pneumococcal surface proteins A and C, hyaluronate lyase, pneumolysin, autolysin, pneumococcal surface antigen A, choline binding protein A, and two neuraminidase enzymes are being investigated as potential vaccine or drug targets. Essentially all of these antigens have been or are being investigated on a structural level in addition to being characterized biochemically. Recently, three-dimensional structures for hyaluronate lyase and pneumococcal surface antigen A became available from X-ray crystallography determinations. Also, modeling studies based on biophysical measurements provided more information about the structures of pneumolysin and pneumococcal surface protein A. Structural and biochemical studies of these pneumococcal virulence factors have facilitated the development of novel antibiotics or protein antigen-based vaccines as an alternative to polysaccharide based vaccines for the treatment of pneumococcal disease. PMID- 11381101 TI - Tetracycline antibiotics: mode of action, applications, molecular biology, and epidemiology of bacterial resistance. AB - Tetracyclines were discovered in the 1940s and exhibited activity against a wide range of microorganisms including gram-positive and gram-negative bacteria, chlamydiae, mycoplasmas, rickettsiae, and protozoan parasites. They are inexpensive antibiotics, which have been used extensively in the prophlylaxis and therapy of human and animal infections and also at subtherapeutic levels in animal feed as growth promoters. The first tetracycline-resistant bacterium, Shigella dysenteriae, was isolated in 1953. Tetracycline resistance now occurs in an increasing number of pathogenic, opportunistic, and commensal bacteria. The presence of tetracycline-resistant pathogens limits the use of these agents in treatment of disease. Tetracycline resistance is often due to the acquisition of new genes, which code for energy-dependent efflux of tetracyclines or for a protein that protects bacterial ribosomes from the action of tetracyclines. Many of these genes are associated with mobile plasmids or transposons and can be distinguished from each other using molecular methods including DNA-DNA hybridization with oligonucleotide probes and DNA sequencing. A limited number of bacteria acquire resistance by mutations, which alter the permeability of the outer membrane porins and/or lipopolysaccharides in the outer membrane, change the regulation of innate efflux systems, or alter the 16S rRNA. New tetracycline derivatives are being examined, although their role in treatment is not clear. Changing the use of tetracyclines in human and animal health as well as in food production is needed if we are to continue to use this class of broad-spectrum antimicrobials through the present century. PMID- 11381100 TI - Viruses in the mammalian male genital tract and their effects on the reproductive system. AB - This review describes the various viruses identified in the semen and reproductive tracts of mammals (including humans), their distribution in tissues and fluids, their possible cell targets, and the functional consequences of their infectivity on the reproductive and endocrine systems. The consequences of these viral infections on the reproductive tract and semen can be extremely serious in terms of organ integrity, development of pathological and cancerous processes, and transmission of diseases. Furthermore, of essential importance is the fact that viral infection of the testicular cells may result not only in changes in testicular function, a serious risk for the fertility and general health of the individual (such as a fall in testosteronemia leading to cachexia), but also in the possible transmission of virus-induced mutations to subsequent generations. In addition to providing an exhaustive account of the data available in these domains, this review focuses attention on the fact that the interface between endocrinology and virology has so far been poorly explored, particularly when major health, social and economical problems are posed. Our conclusions highlight the research strategies that need to be developed. Progress in all these domains is essential for the development of new treatment strategies to eradicate viruses and to correct the virus-induced dysfunction of the endocrine system. PMID- 11381104 TI - Cytokinesis in prokaryotes and eukaryotes: common principles and different solutions. AB - Cytokinesis requires duplication of cellular structures followed by bipolarization of the predivisional cell. As a common principle, this applies to prokaryotes as well as eukaryotes. With respect to eukaryotes, the discussion has focused mainly on Saccharomyces cerevisiae and on Schizosaccharomyces pombe. Escherichia coli and to a lesser extent Bacillus subtilis have been used as prokaryotic examples. To establish a bipolar cell, duplication of a eukaryotic origin of DNA replication as well as its genome is not sufficient. Duplication of the microtubule-organizing center is required as a prelude to mitosis, and it is here that the dynamic cytoskeleton with all its associated proteins comes to the fore. In prokaryotes, a cytoskeleton that pervades the cytoplasm appears to be absent. DNA replication and the concomitant DNA segregation seem to occur without help from extensive cytosolic supramacromolecular assemblies but with help from the elongating cellular envelope. Prokaryotic cytokinesis proceeds through a contracting ring, which has a roughly 100-fold-smaller circumference than its eukaryotic counterpart. Although the ring contains proteins that can be considered as predecessors of actin, tubulin, and microtubule-associated proteins, its macromolecular composition is essentially different. PMID- 11381102 TI - Phi29 family of phages. AB - Continuous research spanning more than three decades has made the Bacillus bacteriophage phi29 a paradigm for several molecular mechanisms of general biological processes, such as DNA replication, regulation of transcription, phage morphogenesis, and phage DNA packaging. The genome of bacteriophage phi29 consists of a linear double-stranded DNA (dsDNA), which has a terminal protein (TP) covalently linked to its 5' ends. Initiation of DNA replication, carried out by a protein-primed mechanism, has been studied in detail and is considered to be a model system for the protein-primed DNA replication that is also used by most other linear genomes with a TP linked to their DNA ends, such as other phages, linear plasmids, and adenoviruses. In addition to a continuing progress in unraveling the initiation of DNA replication mechanism and the role of various proteins involved in this process, major advances have been made during the last few years, especially in our understanding of transcription regulation, the head tail connector protein, and DNA packaging. Recent progress in all these topics is reviewed. In addition to phi29, the genomes of several other Bacillus phages consist of a linear dsDNA with a TP molecule attached to their 5' ends. These phi29-like phages can be divided into three groups. The first group includes, in addition to phi29, phages PZA, phi15, and BS32. The second group comprises B103, Nf, and M2Y, and the third group contains GA-1 as its sole member. Whereas the DNA sequences of the complete genomes of phi29 (group I) and B103 (group II) are known, only parts of the genome of GA-1 (group III) were sequenced. We have determined the complete DNA sequence of the GA-1 genome, which allowed analysis of differences and homologies between the three groups of phi29-like phages, which is included in this review. PMID- 11381106 TI - Abrogation of disease development in plants expressing animal antiapoptotic genes. AB - An emerging topic in plant biology is whether plants display analogous elements of mammalian programmed cell death during development and defense against pathogen attack. In many plant-pathogen interactions, plant cell death occurs in both susceptible and resistant host responses. For example, specific recognition responses in plants trigger formation of the hypersensitive response and activation of host defense mechanisms, resulting in restriction of pathogen growth and disease development. Several studies indicate that cell death during hypersensitive response involves activation of a plant-encoded pathway for cell death. Many susceptible interactions also result in host cell death, although it is not clear how or if the host participates in this response. We have generated transgenic tobacco plants to express animal genes that negatively regulate apoptosis. Plants expressing human Bcl-2 and Bcl-xl, nematode CED-9, or baculovirus Op-IAP transgenes conferred heritable resistance to several necrotrophic fungal pathogens, suggesting that disease development required host cell death pathways. In addition, the transgenic tobacco plants displayed resistance to a necrogenic virus. Transgenic tobacco harboring Bcl-xl with a loss of-function mutation did not protect against pathogen challenge. We also show that discrete DNA fragmentation (laddering) occurred in susceptible tobacco during fungal infection, but does not occur in transgenic-resistant plants. Our data indicate that in compatible plant-pathogen interactions apoptosis-like programmed cell death occurs. Further, these animal antiapoptotic genes function in plants and should be useful to delineate resistance pathways. These genes also have the potential to generate effective disease resistance in economically important crops. PMID- 11381107 TI - Photosystem II single crystals studied by EPR spectroscopy at 94 GHz: the tyrosine radical Y(D)(*). AB - Electron paramagnetic resonance (EPR) spectroscopy at 94 GHz is used to study the dark-stable tyrosine radical Y(D)(*) in single crystals of photosystem II core complexes (cc) isolated from the thermophilic cyanobacterium Synechococcus elongatus. These complexes contain at least 17 subunits, including the water oxidizing complex (WOC), and 32 chlorophyll a molecules/PS II; they are active in light-induced electron transfer and water oxidation. The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with four PS II dimers per unit cell. High-frequency EPR is used for enhancing the sensitivity of experiments performed on small single crystals as well as for increasing the spectral resolution of the g tensor components and of the different crystal sites. Magnitude and orientation of the g tensor of Y(D)(*) and related information on several proton hyperfine tensors are deduced from analysis of angular-dependent EPR spectra. The precise orientation of tyrosine Y(D)(*) in PS II is obtained as a first step in the EPR characterization of paramagnetic species in these single crystals. PMID- 11381105 TI - Identification of GIT1/Cat-1 as a substrate molecule of protein tyrosine phosphatase zeta /beta by the yeast substrate-trapping system. AB - We used a genetic method, the yeast substrate-trapping system, to identify substrates for protein tyrosine phosphatases zeta (PTPzeta/RPTPbeta). This method is based on the yeast two-hybrid system, with two essential modifications: conditional expression of protein tyrosine kinase v-src (active src) to tyrosine phosphorylate the prey proteins and screening by using a substrate-trap mutant of PTPzeta (PTPzeta-D1902A) as bait. By using this system, several substrate candidates for PTPzeta were isolated. Among them, GIT1/Cat-1 (G protein-coupled receptor kinase-interactor 1/Cool-associated, tyrosine-phosphorylated 1) was examined further. GIT1/Cat-1 bound to PTPzeta-D1902A dependent on the substrate tyrosine phosphorylation. Tyrosine-phosphorylated GIT1/Cat-1 was dephosphorylated by PTPzeta in vitro. Immunoprecipitation experiments indicated that PTPzeta D1902A and GIT1/Cat-1 form a stable complex also in mammalian cells. Immunohistochemical analyses revealed that PTPzeta and GIT1/Cat-1 were colocalized in the processes of pyramidal cells in the hippocampus and neocortex in rat brain. Subcellular colocalization was further verified in the growth cones of mossy fibers from pontine explants and in the ruffling membranes and processes of B103 neuroblastoma cells. Moreover, pleiotrophin, a ligand for PTPzeta, increased tyrosine phosphorylation of GIT1/Cat-1 in B103 cells. All these results indicate that GIT1/Cat-1 is a substrate molecule of PTPzeta. PMID- 11381109 TI - The in vivo neuromodulatory effects of the herbal medicine ginkgo biloba. AB - Extracts of Ginkgo biloba leaves are consumed as dietary supplements to counteract chronic, age-related neurological disorders. We have applied high density oligonucleotide microarrays to define the transcriptional effects in the cortex and hippocampus of mice whose diets were supplemented with the herbal extract. Gene expression analysis focused on the mRNAs that showed a more than 3 fold change in their expression. In the cortex, mRNAs for neuronal tyrosine/threonine phosphatase 1, and microtubule-associated tau were significantly enhanced. Hyperphosphorylated tau is the major constituent of the neurofibrillary tangles in the brains of Alzheimer's disease patients. The expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA)-2, calcium and chloride channels, prolactin, and growth hormone (GH), all of which are associated with brain function, were also up-regulated. In the hippocampus, only transthyretin mRNA was upregulated. Transthyretin plays a role in hormone transport in the brain and possibly a neuroprotective role by amyloid-beta sequestration. This study reveals that diets supplemented with Ginkgo biloba extract have notable neuromodulatory effects in vivo and illustrates the utility of genome-wide expression monitoring to investigate the biological actions of complex extracts. PMID- 11381108 TI - Regulation of the stem cell leukemia (SCL) gene: a tale of two fishes. AB - The stem cell leukemia (SCL) gene encodes a tissue-specific basic helix-loop helix (bHLH) protein with a pivotal role in hemopoiesis and vasculogenesis. Several enhancers have been identified within the murine SCL locus that direct reporter gene expression to subdomains of the normal SCL expression pattern, and long-range sequence comparisons of the human and murine SCL loci have identified additional candidate enhancers. To facilitate the characterization of regulatory elements, we have sequenced and analyzed 33 kb of the SCL genomic locus from the pufferfish Fugu rubripes, a species with a highly compact genome. Although the pattern of SCL expression is highly conserved from mammals to teleost fish, the genes flanking pufferfish SCL were unrelated to those known to flank both avian and mammalian SCL genes. These data suggest that SCL regulatory elements are confined to the region between the upstream and downstream flanking genes, a region of 65 kb in human and 8.5 kb in pufferfish. Consistent with this hypothesis, the entire 33-kb pufferfish SCL locus directed appropriate expression to hemopoietic and neural tissue in transgenic zebrafish embryos, as did a 10.4 kb fragment containing the SCL gene and extending to the 5' and 3' flanking genes. These results demonstrate the power of combining the compact genome of the pufferfish with the advantages that zebrafish provide for studies of gene regulation during development. Furthermore, the pufferfish SCL locus provides a powerful tool for the manipulation of hemopoiesis and vasculogenesis in vivo. PMID- 11381103 TI - Natural biology of polyomavirus middle T antigen. AB - "It has been commented by someone that 'polyoma' is an adjective composed of a prefix and suffix, with no root between--a meatless linguistic sandwich" (C. J. Dawe). The very name "polyomavirus" is a vague mantel: a name given before our understanding of these viral agents was clear but implying a clear tumor life style, as noted by the late C. J. Dawe. However, polyomavirus are not by nature tumor-inducing agents. Since it is the purpose of this review to consider the natural function of middle T antigen (MT), encoded by one of the seemingly crucial transforming genes of polyomavirus, we will reconsider and redefine the virus and its MT gene in the context of its natural biology and function. This review was motivated by our recent in vivo analysis of MT function. Using intranasal inoculation of adult SCID mice, we have shown that polyomavirus can replicate with an MT lacking all functions associated with transformation to similar levels to wild-type virus. These observations, along with an almost indistinguishable replication of all MT mutants with respect to wild-type viruses in adult competent mice, illustrate that MT can have a play subtle role in acute replication and persistence. The most notable effect of MT mutants was in infections of newborns, indicating that polyomavirus may be highly adapted to replication in newborn lungs. It is from this context that our current understanding of this well-studied virus and gene is presented. PMID- 11381110 TI - Large conformational changes of the epsilon subunit in the bacterial F1F0 ATP synthase provide a ratchet action to regulate this rotary motor enzyme. AB - The F(1)F(0) ATP synthase is the smallest motor enzyme known. Previous studies had established that the central stalk, made of the gamma and epsilon subunits in the F(1) part and c subunit ring in the F(0) part, rotates relative to a stator composed of alpha(3)beta(3)deltaab(2) during ATP hydrolysis and synthesis. How this rotation is regulated has been less clear. Here, we show that the epsilon subunit plays a key role by acting as a switch of this motor. Two different arrangements of the epsilon subunit have been visualized recently. The first has been observed in beef heart mitochondrial F(1)-ATPase where the C-terminal portion is arranged as a two-alpha-helix hairpin structure that extends away from the alpha(3)beta(3) region, and toward the position of the c subunit ring in the intact F(1)F(0). The second arrangement was observed in a structure determination of a complex of the gamma and epsilon subunits of the Escherichia coli F(1) ATPase. In this, the two C-terminal helices are apart and extend along the gamma to interact with the alpha and beta subunits in the intact complex. We have been able to trap these two arrangements by cross-linking after introducing appropriate Cys residues in E. coli F(1)F(0), confirming that both conformations of the epsilon subunit exist in the enzyme complex. With the C-terminal domain of epsilon toward the F(0), ATP hydrolysis is activated, but the enzyme is fully coupled in both ATP hydrolysis and synthesis. With the C-terminal domain toward the F(1) part, ATP hydrolysis is inhibited and yet the enzyme is fully functional in ATP synthesis; i.e., it works in one direction only. These results help explain the inhibitory action of the epsilon subunit in the F(1)F(0) complex and argue for a ratchet function of this subunit. PMID- 11381111 TI - Effect of COMT Val108/158 Met genotype on frontal lobe function and risk for schizophrenia. AB - Abnormalities of prefrontal cortical function are prominent features of schizophrenia and have been associated with genetic risk, suggesting that susceptibility genes for schizophrenia may impact on the molecular mechanisms of prefrontal function. A potential susceptibility mechanism involves regulation of prefrontal dopamine, which modulates the response of prefrontal neurons during working memory. We examined the relationship of a common functional polymorphism (Val(108/158) Met) in the catechol-O-methyltransferase (COMT) gene, which accounts for a 4-fold variation in enzyme activity and dopamine catabolism, with both prefrontally mediated cognition and prefrontal cortical physiology. In 175 patients with schizophrenia, 219 unaffected siblings, and 55 controls, COMT genotype was related in allele dosage fashion to performance on the Wisconsin Card Sorting Test of executive cognition and explained 4% of variance (P = 0.001) in frequency of perseverative errors. Consistent with other evidence that dopamine enhances prefrontal neuronal function, the load of the low-activity Met allele predicted enhanced cognitive performance. We then examined the effect of COMT genotype on prefrontal physiology during a working memory task in three separate subgroups (n = 11-16) assayed with functional MRI. Met allele load consistently predicted a more efficient physiological response in prefrontal cortex. Finally, in a family-based association analysis of 104 trios, we found a significant increase in transmission of the Val allele to the schizophrenic offspring. These data suggest that the COMT Val allele, because it increases prefrontal dopamine catabolism, impairs prefrontal cognition and physiology, and by this mechanism slightly increases risk for schizophrenia. PMID- 11381112 TI - Rates of nucleotide substitution in sexual and anciently asexual rotifers. AB - The class Bdelloidea of the phylum Rotifera is the largest well studied eukaryotic taxon in which males and meiosis are unknown, and the only one for which these indications of ancient asexuality are supported by cytological and molecular genetic evidence. We estimated the rates of synonymous and nonsynonymous substitutions in the hsp82 heat shock gene in bdelloids and in facultatively sexual rotifers of the class Monogononta, employing distance based and maximum likelihood methods. Relative-rate tests, using acanthocephalan rotifers as an outgroup, showed slightly higher rates of nonsynonymous substitution and slightly lower rates of synonymous substitution in bdelloids as compared with monogononts. The opposite trend, however, was seen in intraclass pairwise comparisons. If, as it seems, bdelloids have evolved asexually, an equality of bdelloid and monogonont substitution rates would suggest that the maintenance of sexual reproduction in monogononts is not attributable to an effect of sexual reproduction in limiting the load of deleterious nucleotide substitutions. PMID- 11381113 TI - Recursive partitioning for tumor classification with gene expression microarray data. AB - Precise classification of tumors is critically important for cancer diagnosis and treatment. It is also a scientifically challenging task. Recently, efforts have been made to use gene expression profiles to improve the precision of classification, with limited success. Using a published data set for purposes of comparison, we introduce a methodology based on classification trees and demonstrate that it is significantly more accurate for discriminating among distinct colon cancer tissues than other statistical approaches used heretofore. In addition, competing classification trees are displayed, which suggest that different genes may coregulate colon cancers. PMID- 11381114 TI - RNA editing activity is associated with splicing factors in lnRNP particles: The nuclear pre-mRNA processing machinery. AB - Multiple members of the ADAR (adenosine deaminases acting on RNA) gene family are involved in A-to-I RNA editing. It has been speculated that they may form a large multicomponent protein complex. Possible candidates for such complexes are large nuclear ribonucleoprotein (lnRNP) particles. The lnRNP particles consist mainly of four spliceosomal subunits that assemble together with the pre-mRNA to form a large particle and thus are viewed as the naturally assembled pre-mRNA processing machinery. Here we investigated the presence of ADARs in lnRNP particles by Western blot analysis using anti-ADAR antibodies and by indirect immunoprecipitation. Both ADAR1 and ADAR2 were found associated with the spliceosomal components Sm and SR proteins within the lnRNP particles. The two ADARs, associated with lnRNP particles, were enzymatically active in site selective A-to-I RNA editing. We demonstrate the association of ADAR RNA editing enzymes with physiological supramolecular complexes, the lnRNP particles. PMID- 11381115 TI - Activation of NF-kappa B is required for hypertrophic growth of primary rat neonatal ventricular cardiomyocytes. AB - The transcription factor NF-kappaB regulates expression of genes that are involved in inflammation, immune response, viral infection, cell survival, and division. However, the role of NF-kappaB in hypertrophic growth of terminally differentiated cardiomyocytes is unknown. Here we report that NF-kappaB activation is required for hypertrophic growth of cardiomyocytes. In cultured rat primary neonatal ventricular cardiomyocytes, the nuclear translocation of NF kappaB and its transcriptional activity were stimulated by several hypertrophic agonists, including phenylephrine, endothelin-1, and angiotensin II. The activation of NF-kappaB was inhibited by expression of a "supersuppressor" IkappaBalpha mutant that is resistant to stimulation-induced degradation and a dominant negative IkappaB kinase (IKKbeta) mutant that can no longer be activated by phosphorylation. Furthermore, treatment with phenylephrine induced IkappaBalpha degradation in an IKK-dependent manner, suggesting that NF-kappaB is a downstream target of the hypertrophic agonists. Importantly, expression of the supersuppressor IkappaBalpha mutant or the dominant negative IKKbeta mutant blocked the hypertrophic agonist-induced expression of the embryonic gene atrial natriuretic factor and enlargement of cardiomyocytes. Conversely, overexpression of NF-kappaB itself induced atrial natriuretic factor expression and cardiomyocyte enlargement. These findings suggest that NF-kappaB plays a critical role in the hypertrophic growth of cardiomyocytes and may serve as a potential target for the intervention of heart disease. PMID- 11381116 TI - Phosphorylation sites of protein kinase C delta in H2O2-treated cells and its activation by tyrosine kinase in vitro. AB - Protein kinase C delta (PKC delta) is normally activated by diacylglycerol produced from receptor-mediated hydrolysis of inositol phospholipids. On stimulation of cells with H(2)O(2), the enzyme is tyrosine phosphorylated, with a concomitant increase in enzymatic activity. This activation does not appear to accompany its translocation to membranes. In the present study, the tyrosine phosphorylation sites of PKC delta in the H(2)O(2)-treated cells were identified as Tyr-311, Tyr-332, and Tyr-512 by mass spectrometric analysis with the use of the precursor-scan method and by immunoblot analysis with the use of phosphorylation site-specific antibodies. Tyr-311 was the predominant modification site among them. In an in vitro study, phosphorylation at this site by Lck, a non-receptor-type tyrosine kinase, enhanced the basal enzymatic activity and elevated its maximal velocity in the presence of diacylglycerol. The mutation of Tyr-311 to phenylalanine prevented the increase in this maximal activity, but replacement of the other two tyrosine residues did not block such an effect. The results indicate that phosphorylation at Tyr-311 between the regulatory and catalytic domains is a critical step for generation of the active PKC delta in response to H(2)O(2). PMID- 11381117 TI - Dual HLA class I and class II restricted recognition of alloreactive T lymphocytes mediated by a single T cell receptor complex. AB - The alloreactive human T cell clone MBM15 was found to exhibit dual specificity recognizing both an antigen in the context of the HLA class I A2 molecule and an antigen in the context of the HLA class II DR1. We demonstrated that the dual reactivity that was mediated via a single clonal T cell population depended on specific peptide binding. For complete recognition of the HLA-A2-restricted specificity the interaction of CD8 with HLA class I is essential. Interestingly, interaction of the CD8 molecule with HLA class I contributed to the HLA-DR1 restricted specificity. T cell clone MBM15 expressed two in-frame T cell receptor (TCR) Valpha transcripts (Valpha1 and Valpha2) and one TCR Vbeta transcript (Vbeta13). To elucidate whether two TCR complexes were responsible for the dual recognition or one complex, cytotoxic T cells were transduced with retroviral vectors encoding the different TCR chains. Only T cells transduced with the TCR Valpha1Vbeta13 combination specifically recognized both the HLA-A2(+) and HLA DR1(+) target cells, whereas the Valpha2Vbeta13 combination did not result in a TCR on the cell surface. Thus a single TCRalphabeta complex can have dual specificity, recognizing both a peptide in the context of HLA class I as well as a peptide in the context of HLA class II. Transactivation of T cells by an unrelated antigen in the context of HLA class II may evoke an HLA class I specific T cell response. We propose that this finding may have major implications for immunotherapeutic interventions and insight into the development of autoimmune diseases. PMID- 11381118 TI - A beta -hairpin structure in a 13-mer peptide that binds alpha -bungarotoxin with high affinity and neutralizes its toxicity. AB - Snake-venom alpha-bungarotoxin is a member of the alpha-neurotoxin family that binds with very high affinity to the nicotinic acetylcholine receptor (AChR) at the neuromuscular junction. The structure of the complex between alpha bungarotoxin and a 13-mer peptide (WRYYESSLEPYPD) that binds the toxin with high affinity, thus inhibiting its interactions with AChR with an IC(50) of 2 nM, has been solved by (1)H-NMR spectroscopy. The bound peptide folds into a beta-hairpin structure created by two antiparallel beta-strands, which combine with the already existing triple-stranded beta-sheet of the toxin to form a five-stranded intermolecular, antiparallel beta-sheet. Peptide residues Y3(P), E5(P), and L8(P) have the highest intermolecular contact area, indicating their importance in the binding of alpha-bungarotoxin; W1(P), R2(P), and Y4(P) also contribute significantly to the binding. A large number of characteristic hydrogen bonds and electrostatic and hydrophobic interactions are observed in the complex. The high affinity peptide exhibits inhibitory potency that is better than any known peptide derived from AChR, and is equal to that of the whole alpha-subunit of AChR. The high degree of sequence similarity between the peptide and various types of AChRs implies that the binding mode found within the complex might possibly mimic the receptor binding to the toxin. The design of the high-affinity peptide was based on our previous findings: (i) the detection of a lead peptide (MRYYESSLKSYPD) that binds alpha-bungarotoxin, using a phage-display peptide library, (ii) the information about the three-dimensional structure of alpha bungarotoxin/lead-peptide complex, and (iii) the amino acid sequence analysis of different AChRs. PMID- 11381119 TI - Blood flow and oxygen delivery to human brain during functional activity: theoretical modeling and experimental data. AB - Coupling of cerebral blood flow (CBF) and cerebral metabolic rate for oxygen (CMRO(2)) in physiologically activated brain states remains the subject of debates. Recently it was suggested that CBF is tightly coupled to oxidative metabolism in a nonlinear fashion. As part of this hypothesis, mathematical models of oxygen delivery to the brain have been described in which disproportionately large increases in CBF are necessary to sustain even small increases in CMRO(2) during activation. We have explored the coupling of CBF and oxygen delivery by using two complementary methods. First, a more complex mathematical model was tested that differs from those recently described in that no assumptions were made regarding tissue oxygen level. Second, [(15)O] water CBF positron emission tomography (PET) studies in nine healthy subjects were conducted during states of visual activation and hypoxia to examine the relationship of CBF and oxygen delivery. In contrast to previous reports, our model showed adequate tissue levels of oxygen could be maintained without the need for increased CBF or oxygen delivery. Similarly, the PET studies demonstrated that the regional increase in CBF during visual activation was not affected by hypoxia. These findings strongly indicate that the increase in CBF associated with physiological activation is regulated by factors other than local requirements in oxygen. PMID- 11381120 TI - Functional switch between motor tracts in the presence of the mAb IN-1 in the adult rat. AB - Fine finger and hand movements in humans, monkeys, and rats are under the direct control of the corticospinal tract (CST). CST lesions lead to severe, long-term deficits of precision movements. We transected completely both CSTs in adult rats and treated the animals for 2 weeks with an antibody that neutralized the central nervous system neurite growth inhibitory protein Nogo-A (mAb IN-1). Anatomical studies of the rubrospinal tracts showed that the number of collaterals innervating the cervical spinal cord doubled in the mAb IN-1- but not in the control antibody-treated animals. Precision movements of the forelimb and fingers were severely impaired in the controls, but almost completely recovered in the mAb IN-1-treated rats. Low threshold microstimulation of the motor cortex induced a rapid forelimb electromyography response that was mediated by the red nucleus in the mAb IN-1 animals but not in the controls. These findings demonstrate an unexpectedly high capacity of the adult central nervous system motor system to sprout and reorganize in a targeted and functionally meaningful way. PMID- 11381121 TI - The chaperone/usher pathways of Pseudomonas aeruginosa: identification of fimbrial gene clusters (cup) and their involvement in biofilm formation. AB - Pseudomonas aeruginosa, an important opportunistic human pathogen, persists in certain tissues in the form of specialized bacterial communities, referred to as biofilm. The biofilm is formed through series of interactions between cells and adherence to surfaces, resulting in an organized structure. By screening a library of Tn5 insertions in a nonpiliated P. aeruginosa strain, we identified genes involved in early stages of biofilm formation. One class of mutations identified in this study mapped in a cluster of genes specifying the components of a chaperone/usher pathway that is involved in assembly of fimbrial subunits in other microorganisms. These genes, not previously described in P. aeruginosa, were named cupA1-A5. Additional chaperone/usher systems (CupB and CupC) have been also identified in the genome of P. aeruginosa PAO1; however, they do not appear to play a role in adhesion under the conditions where the CupA system is expressed and functions in surface adherence. The identification of these putative adhesins on the cell surface of P. aeruginosa suggests that this organism possess a wide range of factors that function in biofilm formation. These structures appear to be differentially regulated and may function at distinct stages of biofilm formation, or in specific environments colonized by this organism. PMID- 11381122 TI - Crystal structure of the SarR protein from Staphylococcus aureus. AB - The expression of virulence determinants in Staphylococcus aureus is controlled by global regulatory loci (e.g., sarA and agr). The sar (Staphylococcus accessory regulator) locus is composed of three overlapping transcripts (sarA P1, P3, and P2, transcripts initiated from the P1, P3, and P2 promoters, respectively), all encoding the 124-aa SarA protein. The level of SarA, the major regulatory protein, is partially controlled by the differential activation of the sarA promoters. We previously partially purified a 13.6-kDa protein, designated SarR, that binds to the sarA promoter region to down-modulate sarA transcription from the P1 promoter and subsequently SarA expression. SarR shares sequence similarity to SarA, and another SarA homolog, SarS. Here we report the 2.3 A-resolution x ray crystal structure of the dimeric SarR-MBP (maltose binding protein) fusion protein. The structure reveals that the SarR protein not only has a classic helix turn-helix module for DNA binding at the major grooves, but also has an additional loop region involved in DNA recognition at the minor grooves. This interaction mode could represent a new functional class of the "winged helix" family. The dimeric SarR structure could accommodate an unusually long stretch of approximately 27 nucleotides with two or four bending points along the course, which could lead to the bending of DNA by 90 degrees or more, similar to that seen in the catabolite activator protein (CAP)-DNA complex. The structure also demonstrates the molecular basis for the stable dimerization of the SarR monomers and possible motifs for interaction with other proteins. PMID- 11381123 TI - Induction of cyclooxygenase-2 by Epstein-Barr virus latent membrane protein 1 is involved in vascular endothelial growth factor production in nasopharyngeal carcinoma cells. AB - Cyclooxygenase-2 (COX-2) is an inducible form of COX and is overexpressed in diverse tumors, raising the possibility of a role for COX-2 in carcinogenesis. In addition, COX-2 contributes to angiogenesis. The Epstein-Barr virus (EBV) oncoprotein, latent membrane protein 1 (LMP1), is detected in at least 70% of nasopharyngeal carcinoma (NPC) and all EBV-infected preinvasive nasopharyngeal lesions. We found that in specimens of LMP1-positive NPC, COX-2 is frequently expressed, whereas LMP1-negative NPC rarely express the enzyme. We next found that expression of LMP1 in EBV-negative nasopharyngeal epithelial cells induced COX-2 expression. Coexpression of IkappaBalpha(S32A/S36A), which is not phosphorylated and prevents NF-kappaB activation, with LMP1 showed that NF-kappaB is essential for induction of COX-2 by LMP1. We also demonstrate that NF-kappaB is involved in LMP1-induced cox-2 promoter activity with the use of reporter assays. Two major regions of LMP1, designated CTAR1 and CTAR2, are signal transducing domains of LMP1. Constructs expressing either CTAR1 or CTAR2 induce COX-2 but to a lesser extent than wild-type LMP1, consistent with the ability of both regions to activate NF-kappaB. Furthermore, we demonstrate that LMP1-induced COX-2 is functional because LMP1 increased production of prostaglandin E(2) in a COX-2-dependent manner. Finally, we demonstrate that LMP1 increased production of vascular endothelial growth factor (VEGF). Treatment of LMP1-expressing cells with the COX-2-specific inhibitor (NS-398) dramatically decreased production of VEGF, suggesting that LMP1-induced VEGF production is mediated, at least in part, by COX-2. These results suggest that COX-2 induction by LMP1 may play a role in angiogenesis in NPC. PMID- 11381124 TI - Ullrich scleroatonic muscular dystrophy is caused by recessive mutations in collagen type VI. AB - Ullrich syndrome is a recessive congenital muscular dystrophy affecting connective tissue and muscle. The molecular basis is unknown. Reverse transcription-PCR amplification performed on RNA extracted from fibroblasts or muscle of three Ullrich patients followed by heteroduplex analysis displayed heteroduplexes in one of the three genes coding for collagen type VI (COL6). In patient A, we detected a homozygous insertion of a C leading to a premature termination codon in the triple-helical domain of COL6A2 mRNA. Both healthy consanguineous parents were carriers. In patient B, we found a deletion of 28 nucleotides because of an A --> G substitution at nucleotide -2 of intron 17 causing the activation of a cryptic acceptor site inside exon 18. The second mutation was an exon skipping because of a G --> A substitution at nucleotide -1 of intron 23. Both mutations are present in an affected brother. The first mutation is also present in the healthy mother, whereas the second mutation is carried by their healthy father. In patient C, we found only one mutation so far the same deletion of 28 nucleotides found in patient B. In this case, it was a de novo mutation, as it is absent in her parents. mRNA and protein analysis of patient B showed very low amounts of COL6A2 mRNA and of COL6. A near total absence of COL6 was demonstrated by immunofluorescence in fibroblasts and muscle. Our results demonstrate that Ullrich syndrome is caused by recessive mutations leading to a severe reduction of COL6. PMID- 11381125 TI - Cloning and characterization of a hormonally regulated rat long chain acyl-CoA synthetase. AB - A previously unidentified gonadotropin-regulated long chain acyl-CoA synthetase (GR-LACS) was cloned and characterized as a 79-kDa cytoplasmic protein expressed in Leydig cells of the rat testis. GR-LACS shares sequence identity with two conserved regions of the LACS and luciferase families, including the ATP/AMP binding domain and the 25-aa fatty acyl-CoA synthetase signature motif, but displays low overall amino acid similarities (23-28%). GR-LACS mRNA is expressed abundantly in Leydig cells of the adult testis and to a lesser degree in the seminiferous tubules in spermatogonia and Sertoli cells. It is also observed in ovary and brain. Immunoreactive protein expression was observed mainly in Leydig cells and minimally in the tubules but was not detected in other tissues. In vivo, treatment with a desensitizing dose of human chorionic gonadotropin caused transcriptional down-regulation of GR-LACS expression in Leydig cells. The expressed protein present in the cytoplasm of transfected cells displayed acyl CoA synthetase activity for long chain fatty acid substrates. GR-LACS may contribute to the provision of energy requirements and to the biosynthesis of steroid precursors and could participate through acyl-CoA's multiple functions in the regulation of the male gonad. PMID- 11381126 TI - Immune-type receptor genes in zebrafish share genetic and functional properties with genes encoded by the mammalian leukocyte receptor cluster. AB - An extensive, highly diversified multigene family of novel immune-type receptor (nitr) genes has been defined in Danio rerio (zebrafish). The genes are predicted to encode type I transmembrane glycoproteins consisting of extracellular variable (V) and V-like C2 (V/C2) domains, a transmembrane region and a cytoplasmic tail. All of the genes examined encode immunoreceptor tyrosine-based inhibition motifs in the cytoplasmic tail. Radiation hybrid panel mapping and analysis of a deletion mutant line (b240) indicate that a minimum of approximately 40 nitr genes are contiguous in the genome and span approximately 0.6 Mb near the top of zebrafish linkage group 7. One flanking region of the nitr gene complex shares conserved synteny with a region of mouse chromosome 7, which shares conserved synteny with human 19q13.3-q13.4 that encodes the leukocyte receptor cluster. Antibody-induced crosslinking of Nitrs that have been introduced into a human natural killer cell line inhibits the phosphorylation of mitogen-activated protein kinase that is triggered by natural killer-sensitive tumor target cells. Nitrs likely represent intermediates in the evolution of the leukocyte receptor cluster. PMID- 11381127 TI - Hyperphosphorylation induces self-assembly of tau into tangles of paired helical filaments/straight filaments. AB - The microtubule-associated protein tau is a family of six isoforms that becomes abnormally hyperphosphorylated and accumulates in the form of paired helical filaments (PHF) in the brains of patients with Alzheimer's disease (AD) and patients with several other tauopathies. Here, we show that the abnormally hyperphosphorylated tau from AD brain cytosol (AD P-tau) self-aggregates into PHF like structures on incubation at pH 6.9 under reducing conditions at 35 degrees C during 90 min. In vitro dephosphorylation, but not deglycosylation, of AD P-tau inhibits its self-association into PHF. Furthermore, hyperphosphorylation induces self-assembly of each of the six tau isoforms into tangles of PHF and straight filaments, and the microtubule binding domains/repeats region in the absence of the rest of the molecule can also self-assemble into PHF. Thus, it appears that tau self-assembles by association of the microtubule binding domains/repeats and that the abnormal hyperphosphorylation promotes the self-assembly of tau into tangles of PHF and straight filaments by neutralizing the inhibitory basic charges of the flanking regions. PMID- 11381128 TI - High efficiency mutagenesis, repair, and engineering of chromosomal DNA using single-stranded oligonucleotides. AB - Homologous DNA recombination is a fundamental, regenerative process within living organisms. However, in most organisms, homologous recombination is a rare event, requiring a complex set of reactions and extensive homology. We demonstrate in this paper that Beta protein of phage lambda generates recombinants in chromosomal DNA by using synthetic single-stranded DNAs (ssDNA) as short as 30 bases long. This ssDNA recombination can be used to mutagenize or repair the chromosome with efficiencies that generate up to 6% recombinants among treated cells. Mechanistically, it appears that Beta protein, a Rad52-like protein, binds and anneals the ssDNA donor to a complementary single-strand near the DNA replication fork to generate the recombinant. This type of homologous recombination with ssDNA provides new avenues for studying and modifying genomes ranging from bacterial pathogens to eukaryotes. Beta protein and ssDNA may prove generally applicable for repairing DNA in many organisms. PMID- 11381129 TI - Preferential interaction of the core histone tail domains with linker DNA. AB - Within chromatin, the core histone tail domains play critical roles in regulating the structure and accessibility of nucleosomal DNA within the chromatin fiber. Thus, many nuclear processes are facilitated by concomitant posttranslational modification of these domains. However, elucidation of the mechanisms by which the tails mediate such processes awaits definition of tail interactions within chromatin. In this study we have investigated the primary DNA target of the majority of the tails in mononucleosomes. The results clearly show that the tails bind preferentially to "linker" DNA, outside of the DNA encompassed by the nucleosome core. These results have important implications for models of tail function within the chromatin fiber and for in vitro structural and functional studies using nucleosome core particles. PMID- 11381130 TI - Direct observation of extension and retraction of type IV pili. AB - Type IV pili are thin filaments that extend from the poles of a diverse group of bacteria, enabling them to move at speeds of a few tenths of a micrometer per second. They are required for twitching motility, e.g., in Pseudomonas aeruginosa and Neisseria gonorrhoeae, and for social gliding motility in Myxococcus xanthus. Here we report direct observation of extension and retraction of type IV pili in P. aeruginosa. Cells without flagellar filaments were labeled with an amino specific Cy3 fluorescent dye and were visualized on a quartz slide by total internal reflection microscopy. When pili were attached to a cell and their distal ends were free, they extended or retracted at rates of about 0.5 microm s( 1) (29 degrees C). They also flexed by Brownian motion, exhibiting a persistence length of about 5 microm. Frequently, the distal tip of a filament adsorbed to the substratum and the filament was pulled taut. From the absence of lateral deflections of such filaments, we estimate tensions of at least 10 pN. Occasionally, cell bodies came free and were pulled forward by pilus retraction. Thus, type IV pili are linear actuators that extend, attach at their distal tips, exert substantial force, and retract. PMID- 11381131 TI - An isoleucine/leucine residue in the carboxyltransferase domain of acetyl-CoA carboxylase is critical for interaction with aryloxyphenoxypropionate and cyclohexanedione inhibitors. AB - cDNA fragments encoding the carboxyltransferase domain of the multidomain plastid acetyl-CoA carboxylase (ACCase) from herbicide-resistant maize and from herbicide sensitive and herbicide-resistant Lolium rigidum were cloned and sequenced. A Leu residue was found in ACCases from herbicide-resistant plants at a position occupied by Ile in all ACCases from sensitive grasses studied so far. Leu is present at the equivalent position in herbicide-resistant ACCases from other eukaryotes. Chimeric ACCases containing a 1000-aa fragment of two ACCase isozymes found in a herbicide-resistant maize were expressed in a yeast ACC1 null mutant to test herbicide sensitivity of the enzyme in vivo and in vitro. One of the enzymes was resistant/tolerant, and one was sensitive to haloxyfop and sethoxydim, rendering the gene-replacement yeast strains resistant and sensitive to these compounds, respectively. The sensitive enzyme has an Ile residue, and the resistant one has a Leu residue at the putative herbicide-binding site. Additionally, a single Ile to Leu replacement at an equivalent position changes the wheat plastid ACCase from sensitive to resistant. The effect of the opposite substitution, Leu to Ile, makes Toxoplasma gondii apicoplast ACCase resistant to haloxyfop and clodinafop. In this case, inhibition of the carboxyltransferase activity of ACCase (second half-reaction) of a large fragment of the Toxoplasma enzyme expressed in Escherichia coli was tested. The critical amino acid residue is located close to a highly conserved motif of the carboxyltransferase domain, which is probably a part of the enzyme active site, providing the basis for the activity of fop and dim herbicides. PMID- 11381132 TI - Maintenance of TCR clonality in T cells expressing genes for two TCR heterodimers. AB - T cell receptor (TCR) allelic exclusion is believed to be primarily mediated by suppression of further recombination at the TCR locus after the expression of a functional TCR protein. Genetic allelic exclusion has been shown to be leaky for the beta chain and, more commonly, for the alpha chain. Here, we demonstrate an additional mechanism by which T cells can maintain monoclonality. T cells from double TCR transgenic mice express only one or the other of the two available TCRs at the cell surface. This "functional allelic exclusion" is apparently due to control of the TCR assembly process because these T cells express RNA and protein for all four transgenic TCR proteins. Lack of cell surface expression of the second TCR may be controlled by a failure to assemble the TCR heterodimer. PMID- 11381133 TI - Distinct functions and cooperative interaction of the subunits of the transporter associated with antigen processing (TAP). AB - The ATP-binding cassette (ABC) transporter TAP translocates peptides from the cytosol to awaiting MHC class I molecules in the endoplasmic reticulum. TAP is made up of the TAP1 and TAP2 polypeptides, which each possess a nucleotide binding domain (NBD). However, the role of ATP in peptide binding and translocation is poorly understood. We present biochemical and functional evidence that the NBDs of TAP1 and TAP2 are non-equivalent. Photolabeling experiments with 8-azido-ATP demonstrate a cooperative interaction between the two NBDs that can be stimulated by peptide. The substitution of key lysine residues in the Walker A motifs of TAP1 and TAP2 suggests that TAP1-mediated ATP hydrolysis is not essential for peptide translocation but that TAP2-mediated ATP hydrolysis is critical, not only for translocation, but for peptide binding. PMID- 11381134 TI - Inhibition of Escherichia coli viability by external guide sequences complementary to two essential genes. AB - Narrow spectrum antimicrobial activity has been designed to reduce the expression of two essential genes, one coding for the protein subunit of RNase P (C5 protein) and one for gyrase (gyrase A). In both cases, external guide sequences (EGS) have been designed to complex with either mRNA. Using the EGS technology, the level of microbial viability is reduced to less than 10% of the wild-type strain. The EGSs are additive when used together and depend on the number of nucleotides paired when attacking gyrase A mRNA. In the case of gyrase A, three nucleotides unpaired out of a 15-mer EGS still favor complete inhibition by the EGS but five unpaired nucleotides do not. PMID- 11381135 TI - Effects of estrogen on growth plate senescence and epiphyseal fusion. AB - Estrogen is critical for epiphyseal fusion in both young men and women. In this study, we explored the cellular mechanisms by which estrogen causes this phenomenon. Juvenile ovariectomized female rabbits received either 70 microg/kg estradiol cypionate or vehicle i.m. once a week. Growth plates from the proximal tibia, distal tibia, and distal femur were analyzed after 2, 4, 6, or 8 weeks of treatment. In vehicle-treated animals, there was a gradual senescent decline in tibial growth rate, rate of chondrocyte proliferation, growth plate height, number of proliferative chondrocytes, number of hypertrophic chondrocytes, size of terminal hypertrophic chondrocytes, and column density. Estrogen treatment accelerated the senescent decline in all of these parameters. In senescent growth plates, epiphyseal fusion was observed to be an abrupt event in which all remaining chondrocytes were rapidly replaced by bone elements. Fusion occurred when the rate of chondrocyte proliferation approached zero. Estrogen caused this proliferative exhaustion and fusion to occur earlier. Our data suggest that (i) epiphyseal fusion is triggered when the proliferative potential of growth plate chondrocytes is exhausted; and (ii) estrogen does not induce growth plate ossification directly; instead, estrogen accelerates the programmed senescence of the growth plate, thus causing earlier proliferative exhaustion and consequently earlier fusion. PMID- 11381136 TI - Localization of GAR transformylase in Escherichia coli and mammalian cells. AB - Enzymes of the de novo purine biosynthetic pathway may form a multienzyme complex to facilitate substrate flux through the ten serial steps constituting the pathway. One likely strategy for complex formation is the use of a structural scaffold such as the cytoskeletal network or subcellular membrane of the cell to mediate protein-protein interactions. To ascertain whether this strategy pertains to the de novo purine enzymes, the localization pattern of the third purine enzyme, glycinamide ribonucleotide transformylase (GAR Tfase) was monitored in live Escherichia coli and mammalian cells. Genes encoding human as well as E. coli GAR Tfase fused with green fluorescent protein (GFP) were introduced into their respective cells with regulated expression of proteins and localization patterns monitored by using confocal fluorescence microscopy. In both instances images showed proteins to be diffused throughout the cytoplasm. Thus, GAR Tfase is not localized to an existing cellular architecture, so this device is probably not used to concentrate the members of the pathway. However, discrete clusters of the pathway may still exist throughout the cytoplasm. PMID- 11381138 TI - Proinflammatory cytokines regulate human glucocorticoid receptor gene expression and lead to the accumulation of the dominant negative beta isoform: a mechanism for the generation of glucocorticoid resistance. AB - Inflammatory responses in many cell types are coordinately regulated by the opposing actions of NF-kappaB and the glucocorticoid receptor (GR). The human glucocorticoid receptor (hGR) gene encodes two protein isoforms: a cytoplasmic alpha form (GRalpha), which binds hormone, translocates to the nucleus, and regulates gene transcription, and a nuclear localized beta isoform (GRbeta), which does not bind known ligands and attenuates GRalpha action. We report here the identification of a tumor necrosis factor (TNF)-responsive NF-kappaB DNA binding site 5' to the hGR promoter that leads to a 1.5-fold increase in GRalpha mRNA and a 2.0-fold increase in GRbeta mRNA in HeLaS3 cells, which endogenously express both GR isoforms. However, TNF-alpha treatment disproportionately increased the steady-state levels of the GRbeta protein isoform over GRalpha, making GRbeta the predominant endogenous receptor isoform. Similar results were observed following treatment of human CEMC7 lymphoid cells with TNF-alpha or IL 1. The increase in GRbeta protein expression correlated with the development of glucocorticoid resistance. PMID- 11381137 TI - In vivo requirement for RecJ, ExoVII, ExoI, and ExoX in methyl-directed mismatch repair. AB - Biochemical studies with model DNA heteroduplexes have implicated RecJ exonuclease, exonuclease VII, exonuclease I, and exonuclease X in Escherichia coli methyl-directed mismatch correction. However, strains deficient in the four exonucleases display only a modest increase in mutation rate, raising questions concerning involvement of these activities in mismatch repair in vivo. The quadruple mutant deficient in the four exonucleases, as well as the triple mutant deficient in RecJ exonuclease, exonuclease VII, and exonuclease I, grow poorly in the presence of the base analogue 2-aminopurine, and exposure to the base analogue results in filament formation, indicative of induction of SOS DNA damage response. The growth defect and filamentation phenotypes associated with 2 aminopurine exposure are effectively suppressed by null mutations in mutH, mutL, mutS, or uvrD/mutU, which encode activities that act upstream of the four exonucleases in the mechanism for the methyl-directed reaction that has been proposed based on in vitro studies. The quadruple exonuclease mutant is also cold sensitive, having a severe growth defect at 30 degrees C. This phenotype is suppressed by a uvrD/mutU defect, and partially suppressed by mutH, mutL, or mutS mutations. These observations confirm involvement of the four exonucleases in methyl-directed mismatch repair in vivo and suggest that the low mutability of exonuclease-deficient strains is a consequence of under recovery of mutants due to a reduction in viability and/or chromosome loss associated with activation of the mismatch repair system in the absence of RecJ exonuclease, exonuclease VII, exonuclease I, and exonuclease X. PMID- 11381139 TI - Targeted mutation of the murine arylhydrocarbon receptor nuclear translocator 2 (Arnt2) gene reveals partial redundancy with Arnt. AB - The ubiquitously expressed basic helix-loop-helix (bHLH)-PAS protein ARNT (arylhydrocarbon receptor nuclear transporter) forms transcriptionally active heterodimers with a variety of other bHLH-PAS proteins, including HIF-1alpha (hypoxia-inducible factor-1alpha) and AHR (arylhydrocarbon receptor). These complexes regulate gene expression in response to hypoxia and xenobiotics, respectively, and mutation of the murine Arnt locus results in embryonic death by day 10.5 associated with placental, vascular, and hematopoietic defects. The closely related protein ARNT2 is highly expressed in the central nervous system and kidney and also forms complexes with HIF-1alpha and AHR. To assess unique roles for ARNT2 in development, and reveal potential functional overlap with ARNT, we generated a targeted null mutation of the murine Arnt2 locus. Arnt2(-/-) embryos die perinatally and exhibit impaired hypothalamic development, phenotypes previously observed for a targeted mutation in the murine bHLH-PAS gene Sim1 (Single-minded 1), and consistent with the recent proposal that ARNT2 and SIM1 form an essential heterodimer in vivo [Michaud, J. L., DeRossi, C., May, N. R., Holdener, B. C. & Fan, C. (2000) Mech. Dev. 90, 253-261]. In addition, cultured Arnt2(-/-) neurons display decreased hypoxic induction of HIF-1 target genes, demonstrating formally that ARNT2/HIF-1alpha complexes regulate oxygen-responsive genes. Finally, a strong genetic interaction between Arnt and Arnt2 mutations was observed, indicating that either gene can fulfill essential functions in a dose dependent manner before embryonic day 8.5. These results demonstrate that Arnt and Arnt2 have both unique and overlapping essential functions in embryonic development. PMID- 11381140 TI - Altered myelopoiesis and the development of acute myeloid leukemia in transgenic mice overexpressing cyclin A1. AB - A mammalian A-type cyclin, cyclin A1, is highly expressed in testes of both human and mouse and targeted mutagenesis in the mouse has revealed the unique requirement for cyclin A1 in the progression of male germ cells through the meiotic cell cycle. While very low levels of cyclin A1 have been reported in the human hematopoietic system and brain, the sites of elevated levels of expression of human cyclin A1 were several leukemia cell lines and blood samples from patients with hematopoietic malignances, notably acute myeloid leukemia. To evaluate whether cyclin A1 is directly involved with the development of myeloid leukemia, mouse cyclin A1 protein was overexpressed in the myeloid lineage of transgenic mice under the direction of the human cathepsin G (hCG) promoter. The resulting transgenic mice exhibited an increased proportion of immature myeloid cells in the peripheral blood, bone marrow, and spleen. The abnormal myelopoiesis developed within the first few months after birth and progressed to overt acute myeloid leukemia at a low frequency ( approximately 15%) over the course of 7-14 months. Both the abnormalities in myelopoiesis and the leukemic state could be transplanted to irradiated SCID (severe combined immunodeficient) mice. The observations suggest that cyclin A1 overexpression results in abnormal myelopoiesis and is necessary, but not sufficient in the cooperative events inducing the transformed phenotype. The data further support an important role of cyclin A1 in hematopoiesis and the etiology of myeloid leukemia. PMID- 11381141 TI - Regulated transposition of a fish transposon in the mouse germ line. AB - Tc1/mariner elements are able to transpose in species other than the host from which they were isolated. As potential vectors for insertional mutagenesis and transgenesis of the mouse, these cut-and-paste transposons were tested for their ability to transpose in the mouse germ line. First, the levels of activity of several Tc1/mariner elements in mammalian cells were compared; the reconstructed fish transposon Sleeping Beauty (SB) was found to be an order of magnitude more efficient than the other tested transposons. SB then was introduced into the mouse germ line as a two-component system: one transgene for the expression of the transposase in the male germ line and a second transgene carrying a modified transposon. In 20% of the progeny of double transgenic male mice the transposon had jumped from the original chromosomal position into another locus. Analysis of the integration sites shows that these jumps indeed occurred through the action of SB transposase, and that SB has a strong preference for intrachromosomal transposition. Analysis of the excision sites suggests that double-strand breaks in haploid spermatids are repaired via nonhomologous end joining. The SB system may be a powerful tool for transposon mutagenesis of the mouse germ line. PMID- 11381142 TI - Abnormal differentiation of epidermis in transgenic mice constitutively expressing cyclooxygenase-2 in skin. AB - In prostanoid biosynthesis, the first two steps are catalyzed by cyclooxygenases (COX). In mice and humans, deregulated expression of COX-2, but not of COX-1, is characteristic of epithelial tumors, including squamous cell carcinomas of skin. To explore the function of COX-2 in epidermis, a keratin 5 promoter was used to direct COX-2 expression to the basal cells of interfollicular epidermis and the pilosebaceous appendage of transgenic mouse skin. COX-2 overexpression in the expected locations, resulting in increased prostaglandin levels in epidermis and plasma, correlated with a pronounced skin phenotype. Heterozygous transgenic mice exhibited a reduced hair follicle density. Moreover, postnatally hair follicle morphogenesis and thinning of interfollicular dorsal epidermis were delayed. Adult transgenics showed a body-site-dependent sparse coat of greasy hair, the latter caused by sebaceous gland hyperplasia and increased epicutaneous sebum levels. In tail skin, hyperplasia of scale epidermis reflecting an increased number of viable and cornified cell layers was observed. Hyperplasia was a result of a disturbed program of epidermal differentiation rather than an increased proliferation rate, as reflected by the strong suppression of keratin 10, involucrin, and loricrin expression in suprabasal cells. Further pathological signs were loss of cell polarity, mainly of basal keratinocytes, epidermal invaginations into the dermis, and formation of horn perls. Invaginating hyperplastic lobes were surrounded by CD31-positive vessels. These results demonstrate a causal relationship between transgenic COX-2 expression in basal keratinocytes and epidermal hyperplasia as well as dysplastic features at discrete body sites. PMID- 11381143 TI - Genetic drift and within-host metapopulation dynamics of HIV-1 infection. AB - Most HIV replication occurs in solid lymphoid tissue, which has prominent architecture at the histological level, which separates groups of productively infected CD4(+) cells. Nevertheless, current population models of HIV assume panmixis within lymphoid tissue. We present a simple "metapopulation" model of HIV replication, where the population of infected cells is comprised of a large number of small populations, each of which is established by a few founder viruses and undergoes turnover. To test this model, we analyzed viral genetic variation of infected cell subpopulations within the spleen and demonstrated the action of founder effects as well as significant variation in the extent of genetic differentiation between subpopulations among patients. The combination of founder effects and subpopulation turnover can result in an effective population size much lower than the actual population size and may contribute to the importance of genetic drift in HIV evolution despite a large number of infected cells. PMID- 11381144 TI - Endothelial cell surface F1-F0 ATP synthase is active in ATP synthesis and is inhibited by angiostatin. AB - Angiostatin blocks tumor angiogenesis in vivo, almost certainly through its demonstrated ability to block endothelial cell migration and proliferation. Although the mechanism of angiostatin action remains unknown, identification of F(1)-F(O) ATP synthase as the major angiostatin-binding site on the endothelial cell surface suggests that ATP metabolism may play a role in the angiostatin response. Previous studies noting the presence of F(1) ATP synthase subunits on endothelial cells and certain cancer cells did not determine whether this enzyme was functional in ATP synthesis. We now demonstrate that all components of the F(1) ATP synthase catalytic core are present on the endothelial cell surface, where they colocalize into discrete punctate structures. The surface-associated enzyme is active in ATP synthesis as shown by dual-label TLC and bioluminescence assays. Both ATP synthase and ATPase activities of the enzyme are inhibited by angiostatin as well as by antibodies directed against the alpha- and beta subunits of ATP synthase in cell-based and biochemical assays. Our data suggest that angiostatin inhibits vascularization by suppression of endothelial-surface ATP metabolism, which, in turn, may regulate vascular physiology by established mechanisms. We now have shown that antibodies directed against subunits of ATP synthase exhibit endothelial cell-inhibitory activities comparable to that of angiostatin, indicating that these antibodies function as angiostatin mimetics. PMID- 11381145 TI - Partial molar volume, surface area, and hydration changes for equilibrium unfolding and formation of aggregation transition state: high-pressure and cosolute studies on recombinant human IFN-gamma. AB - The equilibrium dissociation of recombinant human IFN-gamma was monitored as a function of pressure and sucrose concentration. The partial molar volume change for dissociation was -209 +/- 13 ml/mol of dimer. The specific molar surface area change for dissociation was 12.7 +/- 1.6 nm2/molecule of dimer. The first-order aggregation rate of recombinant human IFN-gamma in 0.45 M guanidine hydrochloride was studied as a function of sucrose concentration and pressure. Aggregation proceeded through a transition-state species, N*. Sucrose reduced aggregation rate by shifting the equilibrium between native state (N) and N* toward the more compact N. Pressure increased aggregation rate through increased solvation of the protein, which exposes more surface area, thus shifting the equilibrium away from N toward N*. The changes in partial molar volume and specific molar surface area between the N* and N were -41 +/- 9 ml/mol of dimer and 3.5 +/- 0.2 nm2/molecule, respectively. Thus, the structural change required for the formation of the transition state for aggregation is small relative to the difference between N and the dissociated state. Changes in waters of hydration were estimated from both specific molar surface area and partial molar volume data. From partial molar volume data, estimates were 25 and 128 mol H2O/mol dimer for formation of the aggregation transition state and for dissociation, respectively. From surface area data, estimates were 27 and 98 mol H2O/mol dimer. Osmotic stress theory yielded values approximately 4-fold larger for both transitions. PMID- 11381147 TI - [Breastfeeding as a vital factor in child health]. AB - Mother's milk is the only adequate nutrition for infants. In the recent years the extensive research documents the numerous advantages of breastfeeding. Especially important is exclusive mothers' milk feeding for infants in the first six months of life. The results of biological and epidemiological studies concerning protective effects of breastfeeding against acute and chronic diseases of early and later childhood were presented. Further needs of breastfeeding promotion in Poland based on the research conducted in the last years, were stressed. PMID- 11381148 TI - [Factors associated with exclusive breastfeeding of infants in Poland]. AB - Factors associated with cessation of exclusive breastfeeding in Poland were studied in 10156 randomly selected infants aged 1-6 months. They were breastfed at the time of data collection. Among them, 4929 (48.5%) infants were exclusively breastfed. In multiple logistic regression analysis, the factors associated with cessation of exclusive breastfeeding were: use of a pacifier, short duration of total breastfeeding and exclusive breastfeeding accepted by mothers, first child in the family, primary education of parents, mother working in farming and non employed father, maternal smoking and maternal age over 34 years. PMID- 11381149 TI - [Models of feeding for healthy children]. AB - The subject of this study was to present dietary guidelines for healthy children recommended by the National Research Institute of Mother and Child. For infants breastfeeding is optimal. All infants should be exclusively breastfed until 6 months of age. Complementary food should be introduced in the 7th month of life according to infants' feeding schedule. Modified cow's milk formulas and complementary food are recommended in non-breastfed infants' nutrition. In all medical institutions infants' feeding should comply with these recommendations. The basis of rational nutrition of young children is high quality and safe food with reference to its nutritional value. Adequate choice of food products, variety of diet and meal frequency is the basis of proper nutrition and proper nutritional habits. Regular eating of 3-4 meals per day, avoiding products rich in fat, sugar, salt, cholesterol, as well as increasing physical activity, are recommended in dietary guidelines for school children. The recommended models should be much more effectively implemented in paediatricians' practice and should be the basis for postgraduate training for doctors, nurses and midwives as well. PMID- 11381150 TI - [Influence of feeding long-chain polyunsaturated fatty acids on children's health]. AB - Milk formulae for term-born infants supplemented with docosahexaenoic acid (DHA) resulted in DHA concentration in the red cells comparable to values observed in breast-fed infants without influence on visual acuity. Prematurely born infants fed with DHA supplemented formulae achieved visual acuity faster than the non supplemented control group. The influence of long-chain polyunsaturated fatty acids (LC-PUFA) on visual and cognitive functions may justify their use if the safety criteria are met. LC-PUFA formulae for low birth weigh infants must contain appropriate concentration of the fatty acids with antyoxidative factors: vitamin E and vitamin C. PMID- 11381151 TI - [Evaluation of the nutritional status and feeding patterns of 3-7 year old children --results of a questionnaire]. AB - Questionnaire studies on feeding patterns and nutritional status of 822 pre school children (age 3-7) in different regions of Poland were performed between June 1999 and January 2000. Nutritional status was evaluated on the basis of anthropometric indices - height, body weight, body mass index (BMI) and BMI z score. The mode of feeding was evaluated on the basis of recall of 24-hours menus. Average daily food rations and their nutritional value were calculated from the questionnaires data. Quantity and number of meals were also evaluated. In a group of 402 boys the average height was between 75-90 percentiles and body mass (50-75 percentiles). In 420 girls the average height was about the 90 percentile (age 3-4) and between 75-90 percentiles (age 5-7). The study showed that normal BMI z-score was found in 75% of children, underweight in 16.5% overweight and obesity in 8.5%. Feeding patterns showed great than differences. It was found that meat, fat, sweets and sugar content in daily food rations exceeded the recommended for pre-school children, but the vegetables intake was lower than recommended. Vitamins A and C deficiency in average daily rations didn't occur. The fat intake provided 36% of total energy, whereas carbohydrates (without lactose and starch) - about 20%. This can have inhibitory effect on microelements (Fe, Zn) absorption. Attention must be paid to the choice of products and the wrong distribution of total energy intake in daily rations. The habits of snack eating between the meals were reported in 91% of children. These snacks consisted of fruits, chips, french fries, sweets and sandwiches. These products provides 232 kcal +/- 171 kcal/day. PMID- 11381152 TI - [Eating patterns of school-aged children and adolescents in Poland - questionnaire investigations]. AB - The aim of the investigations carried out by the National Institute of Mother and Child in school-aged children and adolescents including a group of obese pupils, was to assess the health behaviour concerning eating habits and food consumption. A part of these investigations was carried out in framework of international collaborative studies. In a representative group of pupils in Poland, we found that vegetable, dark bread and milk consumption remained on low level, while the use of soft drinks increased. The number of children who ate breakfast at home and lunch at school decreased especially in girls. We found a statistically evident difference in eating habits between obese adolescents and these with normal body mass, especially concerning regularity of meal consumption. We find, that it is very difficult to change incorrect eating habits in obese adolescents. We observe in our outpatient clinic, that obese adolescents have changed the quantity of food, but they haven't changed their habits. The recommended by dietitions increase of fruits, vegetables and low fat milk consumption in their diet was not generally accepted. PMID- 11381153 TI - [Evaluation of nutritional habits, nutritional status and physical performance in selected group of adolescents]. AB - The purpose of the study was the assessment of nutritional status and nutritional habits as well as physical fitness in a group of 74 adolescents (44 girls and 30 boys) aged 13-19 years, spending their holidays in a recreation centre in Klodzko Valley and the evaluation of correlation coefficients between these parameters. The nutritional status (NS) was evaluated on the basis of body mass index (BMI), skin folds thickness and total body fat (FM). Nutritional habits (NH) were estimated by the method of dietary history with special attention paid to consumption of selected food products and regularity of meals. Physical performance (PhP) of the group was assessed by Harvard's step-test. Evaluation of nutritional habits indicated only a few cases of improper nutrition but about 33% of total examined group followed nutrition guidelines and the scores of NH received by the main part of this group (ca. 66%) were recognised as sufficient. Differences of NS, in adolescents were found, but there was no case of obesity. Significant number of girls (21-55%) showed low values of BMI (<18.4 kg/m2) which could indicate insufficient energy intake. No correlation between NH and NS was however found. Level of physical fitness of the examined group was quite high and strongly correlated with nutritional habits (r = 0.77, p < 0.001). The results proved that the relationship between NH and PhP can be noticed in relatively young persons and that the evaluation of NS of youngsters should include a simple physical performance test. PMID- 11381154 TI - [Importance of proper nutrition before and during pregnancy]. AB - Maternal nutrition before and during pregnancy may influence the course of the pregnancy, foetal development and the child's health in its early and also adult life. Maternal underweight before pregnancy (BMI<19.8 kg/m2) and low pregnancy weight gain may increase the risk of low infant birth weight. There is accumulating evidence that persons who were born with low body mass are more susceptible to insulin-independent diabetes, arterial hypertension, hypercholesterolemia and ischaemic heart disease, than those whose birth weight was normal. Recommendations concerning pregnancy weight gain are discussed. Folic acid deficiency during the periconceptional period may cause neural tube defects in the offspring. Full cover of folic acid requirement is necessary. This may be achieved only by diet supplementation or food fortification. Recommendations concerning folic acid supplementation during the periconceptional period are discussed. Folic acid deficiency during pregnancy may also contribute to the preterm delivery and low infant birth weight. The importance of antioxidant vitamins in the prevention of pregnancy hypertension and the consequences of vitamin A overdosage are discussed. Protective calcium activity against pregnancy hypertension and preterm delivery, the importance of maternal iron supplementation in the prevention of low infant birth weight, and also the problem of maternal zinc deficiency which increases the risk of the low infant birth weight, preterm delivery, malformations, post-term delivery and pregnancy hypertension were discussed as well as the consequences of deficiency of the iodine and n-3 fatty acids in the diet. PMID- 11381155 TI - [Influence of child nutrition on health status during adult years]. AB - The problem of safety of child's nutrition should be considered from the moment of his conception because intrauterine growth may be related to the diseases in adult life: atherosclerosis and coronary heart disease. Strict vegetarian diets are not appropriate for children. The task of proper nutrition in childhood is to prevent adult life diseases and to promote potentially highest quality of life and development of cognitive and intellectual functions, and proper growth. The role of microelements - iron and zinc - is also discussed. PMID- 11381156 TI - [Health - oriented aspects in nutrition of children and adolescents]. AB - The author underlines the necessity of recognition health-oriented aspects in nutrition of children and adolescents for both food producers as well as consumers. It is of great importance not only for growth and development of the young population but also for the health of adults. The author has presented the concept of interrelationship between factors aimed at achieving safe food and proper nutritional guidelines for consumer and the knowledge of genetic and environmental aspects. Educational programmes and research in the field of safe food and proper nutrition sciences should play a crucial role. PMID- 11381157 TI - [Foodstuffs intended for special dietary uses for infants and young children in the light of the directives of the European Union and standards of the Codex Alimentarius FAO/WHO]. AB - Food products for infants and young children are included in the group of foodstuffs intended for special dietary uses. The classification of these products comprises the following groups: newborns infant formulas, infant formulas, processed cereal foods and other baby foods for infants and young children and gluten-free products. In the European Union countries the detailed requirements concerning their qualitative and quantitative composition, labeling, advertising and release on the market as well as the health quality of these products have been laid down in the Council Directive of 3 May 1989 on the approximation of the laws of the Member States relating to foodstuffs intended for particular nutritional uses (89/398/EEC with later amendments) and in the detailed directives. Presently, work is being continued on preparing standards of the Codex Alimentarius FAO/WHO for infant formulas for processed cereal foods for infants and young children and for gluten-free foods. In the successive sessions of the Codex Committee on Nutrition and Foods for Special Dietary Uses drafts of these standards are discussed. In the last session of the Committee in Berlin (19 23.06.2000) it was not possible to reach an agreement of views of the various delegations on the standards or their formulation. The work on them and trials at establishing international recommendations has been deferred to the next meeting. PMID- 11381158 TI - [Water in children's diet]. AB - Water is one of the main dietary components. Its quality plays an important role for the safety of food particularly for infants. For the youngest children the groundwater free of chemical and microbiological pollution and with the proper chemical composition can be used. For infants is recommended natural spring water and natural mineral water of low sodium concentration the measured modulus for the actin gel in the cell body. It is, therefore, unlikely that bone cell processes respond in vivo to fluid shear stress, as proposed in [59]. However, we show that the fluid drag forces on the pericellular matrix which tethers the cell processes to the canalicular wall can produce a 20-100 fold amplification of bone tissue strains in the actin filament bundle of the cell process. PMID- 11381171 TI - Effect of the endothelial surface layer on transmission of fluid shear stress to endothelial cells. AB - Responses of vascular endothelial cells to mechanical shear stresses resulting from blood flow are involved in regulation of blood flow, in structural adaptation of vessels, and in vascular disease. Interior surfaces of blood vessels are lined with a layer of bound or adsorbed macromolecules, known as the endothelial surface layer (ESL). In vivo investigations have shown that this layer has a width of order 1 microm, that it substantially impedes plasma flow, and that it excludes flowing red blood cells. Here, the effect of the ESL on transmission of shear stress to endothelial cells is examined using a theoretical model. The layer is assumed to consist of a matrix of molecular chains extending from the surface, held in tension by a slight increase in colloid osmotic pressure relative to that in free-flowing plasma. It is shown that, under physiological conditions, shear stress is transmitted to the endothelial surface almost entirely by the matrix, and fluid shear stresses on endothelial cell membranes are very small. Rapid fluctuations in shear stress are strongly attenuated by the layer. The ESL may therefore play an important role in sensing of shear stress by endothelial cells. PMID- 11381172 TI - Modulation of A-NK cell rigidity: In vitro characterization and in vivo implications for cell delivery. AB - The delivery of cells to specific regions of the vasculature is a critical step in many therapeutic strategies. These include the packaging of DNA or RNA in cell "vehicles" for delivery to tissues, the reconstitution of differentiated cells to an organ using embryonic stem cells, and the enhancement of the immune response using effector lymphocytes. In most cases, these cells must be injected systemically. Unfortunately, ex vivo manipulation or activation can affect cell visco-elastic properties, making it difficult for the injected cells to traverse capillary beds. Compounding the problem is the fact that common agents used in the laboratory for increasing cell deformability generally have adverse side effects on the therapeutic potential of the cells. Using micropipet aspiration techniques, cytotoxicity assays and in vivo trafficking studies we show that: (1) the rigidity of injected effector cells directly affects resistance to passage through tissue; (2) modulation of cytoskeletal organization can be used to decrease cell rigidity, but can also compromise therapeutic efficacy; and (3) thioglycollate, an agent which does not influence effector lymphocyte cytotoxic activity, reduces cell rigidity and entrapment in the lungs. PMID- 11381173 TI - Alterations in the adhesion behavior of osteoblasts by titanium particle loading: inhibition of cell function and gene expression. AB - Total joint replacement prostheses are required to withstand corrosive environments and sustain millions of loading and articulation cycles during their term of implantation. Wear debris generation has been implicated as one of the primary causes of periprosthetic osteolysis and subsequent implant loosening in total joint replacements. Particulate debris consisting of metals, polyethylene, ceramics, and bone cement have each been shown to provoke a biological response in joint tissues. The major cell types within the interfacial granulomatous fibrous tissues consist of fibroblasts, macrophages, lymphocytes, and foreign body giant cells. Osteoblasts are one of the principal cell types in the bone tissue adjacent to prostheses, maintaining physiologic bone remodeling through the balanced coordination of bone formation and resorption in concert with osteoclasts. To date the phenomenon of osteoblast phagocytosis of titanium particles has been suggested, but has not been sufficiently studied or confirmed. This study seeks to clarify the influence of titanium particles on osteoblast adhesion, deformability, proliferation, and gene expression profile. These studies were accomplished by performing biorheological testing, Northern blot analysis and RNase protection assay. The uptake of metallic particles by the osteoblast resulted in a particle-filament complex formation, which induced a series of variations in cell function. Understanding these variations is critical to expanding our knowledge of implant loosening and elucidating the nature of prosthetic joint failure. This study suggests that the impact of titanium particles on osteoblast function and subsequent implant loosening may have been previously underestimated. PMID- 11381174 TI - Mechanisms for cell activation and its consequences for biorheology and microcirculation: Multi-organ failure in shock. AB - Activation of cells in the vascular compartment causes profound alteration of cell rheological properties with impairment of the microcirculation and initiation of inflammatory reactions. Many cardiovascular diseases have been shown to be associated with cell activation and inflammation. While this situation offers the opportunity for new interventions against the deleterious effects of cell activation, there is the need for a better understanding of the mechanisms that lead to cell activation in the first place. We review here several mechanisms for cell activation in the circulation. We show that in shock, a condition associated with severe forms of cell activation, humoral cell activation factors can be detected in plasma. Further analysis indicates that the source of these humoral activators may be due to the action of pancreatic digestive enzymes in the intestine. Ischemia may serve to open the intestinal brush border and permit entry of pancreatic enzymes into the wall of the intestine to initiate self digestion. In this process low molecular weight but potent cell activators are produced which may escape via the intestinal circulation and the lymphatics into the general circulation. Inhibition of pancreatic enzymes in the lumen of the intestine leads to complete attenuation of humoral activator production as well as many of the deleterious sequelae that accompany shock, such as inflammation and multi-organ failure. We outline a method to carry out biochemical isolation of the cell activators derived from pancreatic enzymes. This analysis shows that there are multiple species of cell activators above and beyond currently known species, many of which have molecular weights below 3000 Da. Identification of the mechanisms that lead to cell activation is an important part to understand the mechanisms that lead to alterations of rheological properties of blood cells in disease and dysfunction of the endothelium and parenchymal cells. Our current evidence suggests that pancreatic digestive enzymes and tissue enzymes may play a central role in humoral activator production. PMID- 11381175 TI - Molecular biology and self-regulatory mechanisms of blood viscosity: a review. AB - Blood viscosity is determined by plasma viscosity, hematocrit, erythrocyte deformability and aggregation. Plasma viscosity and hematocrit are directly regulated by the organism. The molecular biology of the principal determinants of plasma viscosity, i.e., fibrinogen, immunoglobulins, albumin, and lipoproteins is outlined in this work. Hematocrit is regulated by erythropoietin, which is primarily induced by tissue hypoxia. Evidence begins to emerge that autoregulatory mechanisms may be involved in blood viscosity. Viscosity modulates gene transcription for albumin and apolipoproteins in cultured hepatocytes and the erythropoietin response to anemia in rats. Further investigations into these self-regulatory mechanisms in biorheology are, however, needed for a better understanding of blood viscosity regulation in health and disease. PMID- 11381176 TI - Dependence of adhesive behavior of neutrophils on local fluid dynamics in a region with recirculating flow. AB - We have recently described patterns of adhesion of different types of leukocytes downstream of a backward facing step. Here the predicted fluid dynamics in channels incorporating backward facing steps are described, and related to the measured velocities of flowing cells, patterns of attachment and characteristics of rolling adhesion for neutrophils perfused over P-selectin. Deeper (upstream depth 300 microm, downstream depth 600 microm, maximum wall shear stress approximately 0.1 Pa) and shallower (upstream depth 260 microm, downstream depth 450 microm, maximum wall shear stress approximately 0.3 Pa) channels were compared. Computational fluid dynamics (CFD) predicted the presence of vortices downstream of the steps, distances to reattachment of flow, local wall shear stresses and components of velocity parallel and perpendicular to the wall. Measurements of velocities of perfused neutrophils agreed well with predictions, and suggested that adhesion to P-selectin should be possible in the regions of recirculating flow, but not downstream in re-established flow in the high shear channel. When channels were coated with a P-selectin-Fc chimaera, neutrophils were captured from flow and immobilised. Capture showed local maxima around the reattachment points, but was absent elsewhere in the high shear chamber. In the low shear chamber there was depression of adhesion just beyond the reattachment point because of expansion of flow and depletion of neutrophils near the wall. Inside the recirculation zones, adhesion decreased approaching the step because of an increasing, vertically upward velocity component. When channels were coated with P-selectin, neutrophils rolled in all regions, but lifted off the surface as they rolled backwards into low shear regions near the step. Rolling velocity in the recirculation zone was independent of shear stress, possibly because of the effects of vertical lift. We conclude that while local wall shear stress influences adhesive behavior, delivery of cells to the wall and their behavior after capture also depend on components of flow perpendicular to the wall. PMID- 11381177 TI - High viscosity plasma expanders: Volume restitution fluids for lowering the transfusion trigger. AB - Hemorheological studies lead to the axiom that high plasma viscosity is detrimental and that it is beneficial to lower blood viscosity, a precept embodied in the practice of hemodilution, where improved perfusion is attributed to the lowering of blood viscosity. Hemodilution is limited by the transfusion trigger, hemoglobin content of blood of about 7-8 g/dl, which indicates when further volume replacements must restore oxygen carrying capacity with red blood cells (RBC). However, oxygen consumption and delivery are not compromised upon passing this landmark. The reduced blood viscosity does not transmit adequate pressure to the capillaries, causing functional capillary density (FCD) to decrease, jeopardizing organ function through the inadequate extraction of products of metabolism from the tissue by the capillaries. Studies in hemorrhagic shock show that survival is primarily determined by the maintenance of FCD and secondarily by tissue oxygenation. FCD is maintained as hematocrit is reduced beyond the transfusion trigger by increasing plasma viscosity, which transmits systemic pressure to the capillaries and induces vasodilatation through the increased shear stress dependent release of vasodilators. Consequently the transfusion trigger is also a "viscosity trigger" indicating when blood and plasma viscosity are too low. In this condition increasing plasma viscosity is beneficial and extends the transfusion trigger reducing the use of blood transfusions. PMID- 11381178 TI - Modulation of red blood cell aggregation and blood viscosity by the covalent attachment of Pluronic copolymers. AB - Despite many years of research, the physiologic or possible pathologic significance of RBC aggregation remains to be clearly determined. As a new approach to address an old question, we have recently developed a technique to vary the aggregation tendency of RBCs in a predictable and reproducible fashion by the covalent attachment of nonionic polymers to the RBC membrane. A reactive derivative of each polymer of interest is prepared by substitution of the terminal hydroxyl group with a reactive moiety, dichlorotriazine (DT), which covalently bonds the polymer molecule to membrane proteins. Pluronics are block copolymers of particular interest as these copolymers can enhance or inhibit RBC aggregation. Pluronics exhibit a critical micellization temperature (CMT): a phase transition from predominantly single, fully hydrated copolymer chains to micelle-like structures. The CMT is a function of both copolymer molecular mass and concentration. This micellization property of Pluronics has been utilized to enhance or inhibit RBC aggregation and hence to vary low-shear blood viscosity. Pluronic-coated RBCs were prepared using reactive DT derivatives of a range of Pluronics (F68, F88, F98 and F108) and resuspended in autologous plasma at 40% hematocrit. Blood viscosity was measured at a range of shear rates (0.1-94.5 s( 1)) and at 25 and 37 degrees C using a Contraves LS-30 couette low shear viscometer. RBC aggregation and whole blood viscosity was modified in a predictable manner depending upon the CMT of the attached Pluronic and the measurement temperature: below the CMT, RBC aggregation was diminished; above the CMT it was enhanced. This technique provides a novel tool to probe some basic research questions. While certainly of value for in vitro mechanistic studies, perhaps the most interesting application may be for in vivo studies: typically, intravital experiments designed to examine the role of RBC aggregation in microvascular flow require perturbation of the suspending plasma to promote or reduce aggregation (e.g., by the addition of dextran). By binding specific Pluronics to the surface, we can produce RBCs that intrinsically have any desired degree of increased or decreased aggregation when suspended in normal plasma, thereby eliminating many potential artifacts for in vivo studies. The copolymer coating technique is simple and reproducible, and we believe it will prove to be a useful tool to help address some of the longstanding questions in the field of hemorheology. PMID- 11381179 TI - Theoretical and experimental analysis of the sedimentation kinetics of concentrated red cell suspensions in a centrifugal field: determination of the aggregation and deformation of RBC by flux density and viscosity functions. AB - The flow properties of blood are mostly determined using various viscometric approaches, and described in terms of a shear rate or shear stress dependent apparent viscosity. The interpretation of results are rather difficult, especially at low shear rates when particle sedimentation and migration within the viscometer gap are significant. By contrast, analysing the separation process in concentrated RBC suspensions in a centrifugal field also yields information about the viscosity function, including particle-particle interaction and deformation parameters. In this paper, the sedimentation process is approached by means of the theory of kinematic waves and theoretically described by solving the corresponding one-dimensional quasi-linear partial differential equation based on viscosity/flow function as a function of volume concentration. The sedimentation kinetics of rigid spherical RBC suspended in saline and normal RBC suspended in Dx-saline solutions were investigated by means of a separation analyser (LUMiFuge 114). The instrument detects the light transmission over the total length of the cell containing the suspension. During centrifugation the analyser automatically determines the position of the particle free fluid/suspension interface or the sediment by means of a special algorithm. The data obtained with sedimentation of rigid spherical RBC at different volume concentrations demonstrate that, in the case of suspensions rotated in containers of constant cross section, there is good agreement between the theory of kinematic waves developed by Anestis and Schneider (1983) and the results of the experiments. Such good agreement was obtained even though a restrictive one-dimensional model was used to obtain the theoretically derived sedimentation time course. In addition, we describe an algorithm enabling the experimental determination of the viscosity and related flux density function to be made for any suspension. Through this approach, we investigated in detail the rheological behavior of suspended rigid spheres at low Reynolds numbers ranging from 10(-6) to 10(-3). The method here introduced also enabled us to investigate RBC suspensions with respect to the deformability and interactions of the cells by means of the separation analysis. Normal, rigid as well as aggregating RBC exhibited marked differences in the sedimentation kinetics, which were quantified by means of the flux and viscosity functions based on the theory of kinematic waves. PMID- 11381180 TI - Rheological effects of red blood cell aggregation in the venous network: a review of recent studies. AB - It has long been recognized that understanding the rheological properties of blood is essential to a full understanding of the function of the circulatory system. Given the difficulty of obtaining carefully controlled measurements in vivo, most of our current concepts of the flow behavior of blood in vivo are based on its properties in vitro. Studies of blood rheology in rotational and tube viscometers have defined the basic properties of blood and pointed to certain features that may be especially significant for understanding in vivo function. At the same time, differences between in vivo and in vitro systems combined with the complex rheological properties of blood make it difficult to predict in vivo blood rheology from in vitro studies. We have investigated certain flow properties of blood in vivo, using the venular network of skeletal muscle as our model system. In the presence of red blood cell aggregation, venous velocity profiles become blunted from the parabolic as in Poiseuille flow, as pseudo-shear rate (= mean fluid velocity/vessel diameter) is decreased from approximately 100 s(-1) to 5 s(-1). At control flow rates, the short distance between venular junctions does not appear to permit significant axial migration and red cell depletion of the peripheral fluid layer before additional red cells and aggregates are infused from a feeding tributary. Formation of a cell-free plasma layer at the vessel wall and sedimentation in vivo are evident only at very low pseudo-shear rates (<5 s(-1)). These findings may explain in large part observations in whole organs of increased venous resistance with reduction of blood flow. PMID- 11381181 TI - No influence of C-peptide, insulin, and glucagon on blood viscosity in vitro in healthy humans and patients with diabetes mellitus. AB - The influence of the hormones most involved in glucose homeostasis, C-peptide, insulin and glucagon on blood viscosity was tested in vitro. Whole blood (adjusted to haematocrit 45%) from healthy volunteers (n=24) and patients with diabetes mellitus (n=17) was incubated with 10(-7)-10(-10) M C-peptide, insulin or glucagon. None of these peptide hormones, neither at physiological nor at supraphysiological levels, had an influence on high (94.5 s(-1)) or low (0.1 s( 1)) shear rate viscosity. The small group of diabetic patients had a higher plasma viscosity and increased blood viscosity at 94.5 s(-1), which is in agreement with earlier studies, but decreased viscosity at low shear rate. We conclude that C-peptide, insulin and glucagon have no direct effect on blood viscosity in vitro. It is, therefore, unlikely that microvascular disturbances seen with either deficiency or excess of these hormones is due to haemorheological factors. PMID- 11381182 TI - Red blood cell aggregation measurements in whole blood and in fibrinogen solutions by different methods. AB - Red blood cell (RBC) aggregation is an important component of whole blood viscosity, especially at low shear rates, and is the major cause of the non Newtonian flow properties of normal blood. In different laboratories several methods are used to determine RBC aggregation, therefore the standardization of aggregation measurements is especially important for getting comparable results. Aggregation indices of 62 human blood samples were determined by two different methods; LORCA (Laser-assisted Optical Rotational Cell Analyzer) and Myrenne aggregometers and related to plasma fibrinogen concentration. Our results show closer correlation values between RBC aggregation and fibrinogen concentration, when RBC aggregation was measured by LORCA (p<0.001), although correlation could be revealed with Myrenne, when plasma fibrinogen level was less than 4.5 g/l. Correlation coefficient between aggregation indices measured by LORCA and Myrenne aggregometers was also closer below this fibrinogen level. In vitro examination of RBC aggregation in fibrinogen solutions showed that higher fibrinogen concentration results in higher RBC aggregation only up to a certain level similarly to that found in human whole blood samples. Our results show that plasma fibrinogen level has an important, concentration dependent effect on RBC aggregation in human blood, but above a certain level it may not cause further aggregate formation. PMID- 11381183 TI - Membrane lipid components associated with increased filterability of erythrocytes from long-distance runners. AB - Deformability of red blood cells and the lipid composition of their plasma membranes were studied in ten long-distance runners, in comparison with ten sedentary matched controls. Mean corpuscular volume and corpuscular hemoglobin concentration did not differ between these groups, while the hemoglobin concentration and hematocrit were lower in the former. Deformability was measured in terms of the erythrocyte suspension filterability through 5 microm microchannels, using an array flow analyzer, and was significantly elevated in athletes. Analysis of the membrane phospholipid composition of red blood cells revealed an increased ratio of phosphatidylcholine to phosphatidylethanolamine and a decreased ratio of cholesterol to total phospholipids in athletes. The length and saturation levels of fatty acid acyl chains were unchanged. Multiple regression analysis showed clearly that these alterations of membrane lipids correlate with increased deformability and thus favor circulation in the microvasculature in long-distance runners. PMID- 11381184 TI - Effect of high-dose progestogen on hemostatic properties of blood in patients with endometrial cancer. AB - Fifteen cases of endometrial cancer were administered daily doses of 600 mg of MPA after surgery to prevent the recurrence of cancer. The initiation times of coagulation (time necessary for fibrin network formation) were measured with a highly sensitive damped oscillation rheometer and compared with those of 15 control patients who were not administered MPA. Biochemical studies of blood coagulation and fibrinolysis were also done. The initiation times of coagulation were 19.0+/-1.8 minutes (min mean +/- standard deviation) after 3-6 months and 16.0+/-2.0 min after 9-12 months of MPA administration, both times being significantly shorter compared with the controls (24.0+/-2.5 min). Hematocrit values, platelet counts and fibrinogen levels were similar between the two groups. Activated partial thromboplastin time (APTT) was significantly decreased and antithrombin III activity (AT III), thrombin-antithrombin complex (TAT), plasminogen level, plasmin-alpha(2) plasmin inhibitor complex level (PIC) and the fibrin degradation product level (FDP) were significantly increased in the MPA group compared with the control group. Accelerated coagulation of blood was definitely induced by high-dose MPA but antithrombin and fibrinolytic activities were also induced, and, thus, thromboembolic complications were prevented. PMID- 11381185 TI - Light and scanning-electron microscopic examination of coronary stents in a perfusion-model. Effects of an NO-donor/quantitative analysis. AB - Does a NO-donor (Corvaton, Aventis, France) attenuate the adherence and the aggregation of platelets in a closed-loop perfusion modell with or without the implantation of coronary stents? SIN-1, the active metabolite of molsidomine, exerted a strong influence on the interaction of platelets with the surfaces of stents. When SIN-1 was added the adherence of platelets to the surfaces of stents was markedly reduced. The sites were most of the platelets adhered to, also changed. More platelets adhered to the stent backbone and less to the free modular stent parts. The localisation of adherent platelets could easily be shown by light microscopy. The aggregation of platelets, accompanied by a spheroidic shape change, could be demonstrated by scanning electron microscopy on the addition of an NO-donor, not only a reduction of platelet adherence was realised but also the reduction of platelet aggregation. PMID- 11381188 TI - Association between the endogenous retrovirus HRES-1 and multiple sclerosis in the United Kingdom--evidence of genetically different disease subsets? AB - In the present study we determined the frequencies of four haplotypes of the human T-cell lymphotropic virus-related endogenous sequence, HRES-1, in 110 multiple sclerosis (MS) patients and 100 healthy control subjects from the United Kingdom. We found evidence of an association between this endogenous retrovirus and MS (p < 0.01), in particular reflecting an increased frequency of HRES-1 haplotype 1 in the group of patients. There was no significant difference in the distribution of HRES-1 haplotypes between relapsing-remitting MS and the primary progressive form of the disease. The odds ratio for HRES-1 haplotype 1 and MS did not differ significantly between individuals positive for HLA-DR2 and DR2 negative individuals. Comparison of the observations from the present study with previous results implicated HRES-1 as a marker of genetic heterogeneity in MS. PMID- 11381186 TI - The effect of acenocoumarol on hemorheological parameters. AB - The effect of acenocoumarol on hemorheological variables was measured in 35 non valvular chronic atrial fibrillation patients before starting oral anticoagulant therapy (basal) and one and two months after beginning treatment (INR-2,3). Fibrinogen increased significantly from the basal situation: 332+/-99 mg/dl to 386+/-96 mg/dl in the second month (p<0.05). However, this small increase in fibrinogen is not large enough to mediate other rheological changes, and whole blood filterability, blood viscosity, plasma viscosity and erythrocyte deformability and aggregability remained unchanged after treatment. These results suggest that acenocoumarol does not affect rheological parameters and can therefore be used as a "neutral drug" for rheological studies in cardiovascular patients under oral anticoagulant therapy. PMID- 11381189 TI - Clinical significance and prognostic value of CA72-4 compared with CEA and CA19-9 in patients with gastric cancer. AB - Carcinoembryonic antigen (CEA) and CA 19-9 are both widely used in the follow up of patients with gastrointestinal cancer. More recently another tumor marker, named CA 72-4 has been identified and characterized using two different monoclonal antibodies B72.3 and CC49. Several reports evaluated CA 72-4 as a serum tumor marker for gastric cancer and compared its clinical utility with that of CEA or CA 19-9; few reports concerned its prognostic value. In the present study, CA 72-4 is evaluated and compared with CEA and CA 19-9 in various populations of patients with gastric cancer and benign disease; for 52 patients with gastric adenocarcinoma and 57 patients without neoplastic disease CEA, CA 19 9 and CA 72-4 were evaluated before treatment. Sensitivity of the tumor markers CA 72-4, CA 19-9 and CEA at the recommended cut-off level in all 52 patients were 58%, 50% the sensitivity increased to 75%. of these markers, for non metastatic patients, multivariate analyses indicated that none of the markers were significant, when adjusted for gender and age (which were indicators of poor prognosis); patients with abnormal values of CA72-4 tended to have shorter survival than patients with normal values (p<0.07). In the metastatic population, only high values of CA19-9 (p<0.02) and gender (women) p<0.03) were indicators of poor prognosis in univariate analysis; multivariate analysis revealed that both CA72-4 (p=0.034) and CA19-9 p=0.009), adjusted for gender were independent prognostic factors. However, CA72-4 lost significance (p=0.41) when adjusted for CA19-9 and gender, indicating that CA19-9 provides more prognostic information than CA72-4. When limited to the metastatic male population with normal values of CA 19-9 and CEA, CA 72-4 pretherapeutic positive levels were associated with a worse prognosis (p<0.005). In conclusion, this study suggests that the addition of CA 72-4 to CEA and/or CA 19-9 could improve sensitivity in gastric cancer. The prognostic role of this marker is not yet clearly demonstrated but its usefulness in the monitoring of gastric cancer should be taken into account. PMID- 11381190 TI - Biochemical changes in patients with combined chronic schistosomiasis and viral hepatitis C infections. AB - This study was undertaken to assess the biochemical changes induced in chronic schistosomiasis and/or chronic HCV, as well as to pinpoint the most significant parameters which could be used as dependable indices for the differentiation of single and coupled infections with or without liver cirrhosis. The selected patients were allocated into 2 broad groups: GrII (Schistosomiasis) which was subdivided into 3 subgroups: GrII(a) schistosomal patients with hepatosplenomegaly; GrII(b) hepatosplenic schistosomal patients with decompensated liver cirrhosis; GrII(c) schistosomal patients with no organomegaly. GrIII (Combined) comprised 2 subgroups: GrIII(a) schistosomal-HCV infection with decompensated liver cirrhosis; GrIII(b) schistosomal-HCV infection without liver cirrhosis. For statistical comparison normal healthy subjects were taken as a reference group (Gr I). Results showed that schistosomal patients without organomegaly manifested non significant changes in all studied parameters compared to normal controls. Highly significant elevations in serum ALT, AST, ALP and GGT activities were recorded in all other subgroups but the highest levels are reported in GrIIb. AST/ALT and direct/indirect bilirubin ratios were highest in GrIIIa (1.17+/-0.26, 1.54 +/- 0.37, respectively). Serum total protein and albumin levels showed the highest reduction (33 and 59%) concomitantly with the highest increase in gamma-globulin level (75%) in GrIII(a). Blood total iron was significantly reduced in GrII(a,b) (15.6 and 12%) (8.8%) bilirubin, GGT and AST in this order are good discriminators between the different subgroups in GrII. On the other hand, ALT, AST, albumin, ALP, GGT, protein and direct bilirubin are the most significant indices to differentiate chronic schistosomiasis and the combined group with/or without liver cirrhosis. PMID- 11381191 TI - Human surfactant protein--A gene locus for genetic studies in the Finnish population. AB - Lung surfactant lowers the surface tension but surfactant proteins also have other functions. Surfactant protein A (SP-A) has a well-defined role in innate immunity. The gene locus for human SP-A genes is in chromosome 10q21 through q24 and consists of two highly homologous functional SP-A genes (SP-A1 and SP-A2) and a pseudogene. Several alleles that differ by a single amino acid have been identified for both SP-A genes. The SP-A gene locus has been shown to be sufficiently polymorphic for genetic studies in the American population. In this study, we analysed the SP-A allele frequencies in a Finnish population (n = 790) and found them to differ from the frequencies observed in US. Furthermore, we describe several new alleles for both SP-A genes. The heterozygosity indices and polymorphism information content values ranged between 0.50--0.62 indicating that SP-A gene locus is polymorphic enough for studies associating the locus with pulmonary diseases. PMID- 11381192 TI - Screening of dystrophin gene deletions in Egyptian patients with DMD/BMD muscular dystrophies. AB - Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are allelic disorders caused by mutations within the dystrophin gene. Our study has identified 100 Egyptian families collected from the Human Genetics Clinic, National Research Center, Cairo. All cases were subjected to complete clinical evaluation pedigree analysis, electromyography studies, estimation of serum creatine phosphokinase enzyme (CPK) levels and DNA analysis. Multiplex PCR using 18 pairs of specific primers were used for screening of deletion mutations within the dystrophin gene. A frequency of 55% among the families. Sixty per cent of detected deletions involved multiple exons spanning the major or the minor hot spot of the dystrophin gene. The remainder 40% which mainly involved exon 45. Comparing these findings with frequencies of other countries it was found that our figures fall within the reported range of 40%- distribution of deletions in our study and other different studies was variable and specific ethnic differences do not apparently account for specific deletions. In addition this study concluded that employment of the 18 exon analysis is a cost effective and a highly accurate (97% to launch a nationwide program. PMID- 11381193 TI - Contribution of flow cytometry to acute leukemia classification in Tunisia. AB - The precision of immunological characterization of leukemias was improved by a certain number of technical innovations, particularly hybridoma production and standardization, resulting in monoclonal antibodies and definition of recognised cellular antigens (designated by CD: Cluster of Differentiation). The aim of this work was to determine the immunophenotyping profile of patients with leukemia, by means of a flow cytometric method: 66 blood samples coming from leukemic persons in the Sahel region were studied by flow cytometry, using about thirty monoclonal antibodies all marked with a fluorochrome, in one or two colour systems to assess their distribution according to type (lymphoid B or T / myeloid) and age, and to search for possible co-expressions of markers of different lineages. The marked preponderance of childhood B-ALL in our series is, at least partly, attributable to the age distribution of the Tunisian population. In agreement with studies from other countries, the majority of AML cases occurred among adults. A high proportion of AML cases in our series co-expressed markers of other lineages. Overall, accurate classification of acute leukemias was possible from a simple peripheral blood sample in 62 of 66 cases (93.9%). PMID- 11381194 TI - HLA-association in patients with intolerance to mercury and other metals in dental materials. AB - A group of selected 25 patients with serious intolerance to heavy metals used for dental restoration were examined for HLA antigens. A significant increase for HLA -- B37, B47 and DR4 was found. The value of the relative risk is not significant after correction for the number of antigens tested and therefore further studies of more patients are needed. PMID- 11381195 TI - Rh system and intrauterine growth. Interaction with season of birth. AB - Based on the hypothesis that maternal-fetal genetic differences in membrane transport and signal transduction may influence intrauterine development, the recent acquisition on transport function of Rh protein prompted us to study the relationship between joint maternal-fetal Rh phenotype and birth weight. Considering that metabolic effect of maternal-fetal competition could be amplified by environmental conditions, we have investigated possible seasonal effects on such relationship. We have studied 5291 infants born in Sardinia in the period January 1993--December 1996 and 984 infants born in Rome during 1996. In Rh(-) mothers there is a significant association between season of birth and birth weight that shows the highest mean value in infants born in autumn (i.e. conceived in winter). The association is much more evident in male than in female infants. In male infants from Rh(-) mothers, the association between birth weight and season is significant in Rh(+) male newborns only. Recent observations by our group in NIDDM suggest that glucose transport in RBC may be related to D protein, thus we propose an interpretation of the present observation in terms of transport function. When the density of D protein in the infant is greater than in the mother, the balance is in favour of the infant who may attain a significant developmental advantage when conceived in the cold season. PMID- 11381196 TI - The significance of PSA/IGF-1 ratio in differentiating benign prostate hyperplasia from prostate cancer. AB - The importance of insulin-like growth factor 1 (IGF-1) in human serum for the early diagnosis of prostate cancer is controversial. The IGF-1/PSA ratio may improve the performance of prostate specific antigen (PSA) as a prostate cancer marker. IGF-1, along with PSA and free PSA concentration, was measured in the serum of 34 patients with prostate cancer and in 131 patients with benign prostatic hyperplasia (BPH). Although IGF-1 concentration did not significantly differ between the groups, PSA/IGF-1 ratio could clearly distinguish the two groups. In patients with cancer but not in patients with BPH, IGF-1 concentration correlated with PSA and free PSA. The values of PSA and free PSA correlated with each other for both groups. Receivers Operating Curve (ROC) analysis indicated a better sensitivity to specificity ratio for PSA/IGF-1 than for PSA or Free/Total (F/T) PSA. PMID- 11381197 TI - Evidence for an association between the SRD5A2 (type II steroid 5 alpha reductase) locus and prostate cancer in Italian patients. AB - We have investigated the contributions of three polymorphic markers in the SRD5A2 gene to prostate cancer in a group of Italian patients. We have genotyped cases and controls for a polymorphic (TA)n dinucleotide repeat and two functional substitutions, A49T and V89L, substituting respectively alanine with threonine at codon 49, and valine to leucine at codon 89. We found a substantially increased but not significant risk associated with the 49T mutation and a reduction of risk for the V89L substitution. In conclusion, we report on preliminary evidence for both increased and decreased risk associated with separate markers at this locus. PMID- 11381198 TI - Sex steroid hormones in serum and tissue of benign and malignant breast tumor patients. AB - The ability of breast tumors to synthesize sex steroid hormones is well recognized and their local production is thought to play a role in breast cancer development and growth. The aim of this study was to estimate local intra-tumoral and circulating levels of Estrone (E1), Estrone Sulfate (E1S), Estradiol (E2), Estriol (E3), and Testosterone (T) in 33 pre- and postmenopausal women with primary breast cancer in comparison to 12 pre- and postmenopausal women with benign breast tumors. The mean levels of the studied sex hormones were higher in serum and tumor tissue of breast cancer women than those with benign breast tumors apart from Testosterone which showed a significant decrease in pre- and postmenopausal women with breast cancer (P<0.001for follicular phase, P<0.05 for luteal phase, and P<0.005 for postmenopausal). The levels of the five hormones were significantly higher intra-tumoral than in serum of both benign and malignant breast tumor women with E1S as the predominant estrogen. There was only a positive significant correlation between serum and tumor tissue levels of E1 (rs=0.52, P<0.05 for follicular; rs=0.63, P<0.05 for luteal and rs=0.58, P<0.05 for postmenopausal) and a significant correlation between serum and tumor tissue of T (rs=0.64, P<0.05 for follicular; rs=-0.51, P<0.05 for luteal and rs=-0.81, P<0.04 for postmenopausal). PMID- 11381199 TI - Study of apolipoprotein E polymorphism in normal healthy controls from northern India. PMID- 11381200 TI - Inflammatory bowel disease: Are there gender differences in the genetics of signal transduction? A preliminary study of cytosolic low molecular weight protein tyrosine phosphatase. AB - The phenotype of cytosolic Low Molecular Weight Protein Tyrosine Phosphatase (cLMWPTP or ACP1), an enzyme involved in signal transduction of insulin, PDGF and T-cell receptors, has been determined in 71 patients with Crohn's Disease (CD: 37 males and 34 females), 49 patients with Ulcerative Colitis (UC: 27 males and 22 females) and 358 consecutive newborns (194 males and 164 females). cLMWPTP phenotypes showing a high concentration of F isoforms are associated with CD in females and with UC in males. Since PTPases counteract the effects of protein tyrosines kinases, a high concentration of F isoform of cLMWPTP may influence the mucosal response to pathogenic factors, increasing susceptibility to CD in females and to UC in males. PMID- 11381201 TI - Linking genotype with phenotype in inflammatory bowel disease--Will we ever have reagent standard patients? PMID- 11381202 TI - Abstracts of the first scientific workshop of the Early Detection Research Network. September 2000. PMID- 11381203 TI - A hemodynamic analysis for determining critical hematocrit in transfusions for sickle cell patients. AB - Because of complicated rheological issues related with sickle cell blood, there exists a practical need to rationally determine critical hematocrit in transfusions for sickle cell patients. In this research, two major effects, i.e. oxygen concentration and hematocrit, are considered in a theoretical hydrodynamic model incorporating oxygen transport and lubrication theory. The pressure drop depends on the compliance of red blood cells, which changes with oxygen concentration in sickle red cells. Under the assumption that after transfusion, the upper limit of the local resistance of a capillary should not surpass that in the same capillary of normal blood flow, critical hematocrit values with different exchange percentage rates are determined. The current clinical ceiling value of 35 percent for total hematocrit after the transfusion, for the first time, finds its analytical reasoning through mathematical modeling. Plots of critical hematocrit values versus the transfusion exchange percentage are provided in the paper. PMID- 11381205 TI - Analysis of mechanical stresses within the alveolar septa leading to pulmonary edema. AB - Mechanical ventilation has been associated with pulmonary edema in the clinical setting, but the pathophysiological mechanisms of this process have not been clearly defined. Experimental studies have shown that high transpulmonary pressures resulting from ventilation may damage the capillary walls, thereby leading to edema. Knowledge of the stress distribution within the alveolar septa would be an important step in understanding this phenomenon. A newly developed saline-filled alveolar sac model was utilized for analysis of septal stresses in young and aging healthy lungs, in order to examine their vulnerability to pulmonary edema during ventilation. Significant stress concentrations were shown to develop near highly curved regions (small local radii of less than 4 mum in a lung inflated to 80% could be as high as 25 times that of average septal stresses. The combination of elevated stress sites that are formed in the stiffer parenchyma of the aging lung, together with the cyclic loading of ventilation, may explain the gaps and breaks previously observed in pulmonary edema. PMID- 11381204 TI - Educational simulation of the electroencephalogram (EEG). AB - We describe a model for simulating a spontaneous electroencephalogram (EEG) and for simulating the effects of anesthesia on the EEG, to allow anesthesiologists and EEG technicians to learn and practice intraoperative EEG monitoring. For this purpose, we developed a linear model to manipulate the amplitude of the activity in each of the traditional EEG frequency bands. Burst suppression patterns are simulated by manipulating an overall gain. To demonstrate the model feasibility, model parameters for thiopental and isoflurane were estimated guided by published data on the EEG effects of these anesthetic drugs. Using these estimates, EEG time signals were simulated for isoflurane at various partial pressures, and for bolus intravenous doses of thiopental. Comparison with actual recorded EEG signals showed that the changes produced by isoflurane and thiopental in the simulated signals are very similar to the changes in the actual signals, which was confirmed by two clinicians with experience and routine practice in intraoperative EEG monitoring. PMID- 11381206 TI - Time-frequency analysis of breathing signals: in vitro airway model. AB - Sound signals of respiratory airflow represent summations of acoustic waves of various frequencies, which basically depend on the characteristics of the flow and on those of the surrounding tissue. This study was designed to examine the capability of time-frequency distribution (TFD) of respiratory signals in order to differentiate between unobstructed and obstructed upper airways. In order to investigate the TFD characteristics of defined upper airway geometry we conducted a controlled basic study in a laboratory system with an in vitro isolated airway model, which was either unobstructed or had concentric obstructions of various degrees at different locations along the tube. Pressure fluctuations were acquired with a microphone proximal to the airway opening. A short-term Fourier transform was used to study the TFDs of these signals. The results of the in vitro study showed that the energy of the higher frequencies increased for relatively small incremental changes in: i) reduction of the lumen cross-section, ii) decrease of distance from measurement site to obstruction, and iii) increase of breathing effort. Further development of this method may lead to noninvasive clinical techniques for early diagnosis of upper airway obstructions. PMID- 11381207 TI - Application criteria for infrared ear thermometers in pediatric surgery. AB - Fever is an important and sensitive indicator of infectious diseases in children. For the past decades, measurement of body temperature in routine medical practice was limited to oral, rectal and axillary sites. In infants and children, however, temperature measurements are further limited to the rectal and axillary sites due to technical and clinical considerations. In the field of pediatric surgery, pathological and surgical conditions of the ano-rectal area often further restrict the application of conventional rectal temperature measurements. The application of tympanic temperature measurements in such pediatric surgical pathologies provides a reliable alternative to conventional rectal temperature measurements. The pediatric surgical clinico-pathological states where ear temperature measurements are the only mode of accurate temperature determination have been identified. Tympanic thermometry is well tolerated by children and predicts temperature with relative accuracy within a few seconds. PMID- 11381208 TI - The effect of fluid shear stress on ICAM-1 expression of rat brain microvascular endothelial cells. AB - Intercellular adhesion molecule-1 (ICAM-1) is an adherence molecule that is an important factor in many pathophysiological processes such as atherosclerosis, thrombosis and inflammation. It is secretion of endothelial cells by a variety of biochemical stimulations. But hemodynamic forces can also induce various functional changes in vascular endothelium. Some researches have proved that shear stress can modulate the expression of ICAM-1. But most of them examine the regulation of expression of ICAM-1 in human umbilical vein endothelial cells. There is no detail on the effect of shear stress (SS) on ICAM-1 expression of microvascular endothelial cells (RBMECs). In this experiment, we use cultured rat brain microvascular endothelial cells (RBMECs). By using the parallel plate flow chamber method, we give two magnitudes of lamminar shear stresses (0.2 dyn/cm2, 0.4 dyn/cm2) for different perieods of time on the slides of cells. Immunostaining method and image analysis shows a specific upregulation in ICAM-1 expression on RBMECs, which is different from endothelial cells of other species or vascular beds. Expression of ICAM-1 is increased 0.5h after the onset of SS, and reached its highest level 4h after onset of SS, then declines after that. The effect is time-dependent, not force magnitude-dependent. Endothelial cell surface expression of ICAM-1 in the supernatants of RBMECs exposed to SS was not modified excluding the possibility that RBMECs exposed to SS synthesize factors that upregulate ICAM-1. The experiment data are relevant to the current understanding of basic mechanisms that explain the signal transudation pathway occurring inside the endothelial cells under the effect of SS. PMID- 11381209 TI - The giant stirs. PMID- 11381210 TI - Newsfronts. PMID- 11381211 TI - Usda draft policy on potentially dangerous animals. PMID- 11381212 TI - An adequate av? PMID- 11381213 TI - Ocular discharge in a guinea pig. PMID- 11381214 TI - Part-time employees in the research facility. AB - As the "eyes and ears" of the biomedical research team, laboratory animal technicians perform functions essential to maintaining the health of the laboratory animals, and advocate their welfare. The author describes a program that allows part-time employees to perform some of the laboratory animal technician responsibilities when facilities are understaffed and low on funds. PMID- 11381215 TI - OLAW and APHIS: Common Areas of Noncompliance. AB - The Director of the Compliance Oversight Division, Office of Laboratory Animal Welfare, NIH, and the Deputy Administrator of USDA/APHIS Animal Care discuss areas of noncompliance with PHS Policy and the AWA. PMID- 11381216 TI - Red-carpet rodent care: making the most of dollars and sense in the animal facility. AB - Cutting-edge biomedical research programs have entered an era in which phenotypic characterizations for genetically altered rodents can facilitate appropriate care. The veterinary care requirements necessary to support such animal models can include the procedures already adapted as standard practice in companion animal hospitals, and can decrease data variability while increasing survival rates. PMID- 11381218 TI - It's a small world after all. PMID- 11381217 TI - Keeping health and safety first. PMID- 11381219 TI - Newsfronts. PMID- 11381220 TI - Iccvam authorization act. PMID- 11381221 TI - Blind to distress? PMID- 11381222 TI - Laboratory-Acquired Lymphadenopathy in a Veterinary Pathologist. PMID- 11381223 TI - Assessing animal care and use programs internationally. AB - As international collaborations become commonplace, new issues arise concerning the potentially variable quality of laboratory animals. The Associate Director and the Director of AAALAC International discuss efforts to harmonize facility procedures and practices of animal care and use globally. PMID- 11381224 TI - Biosafety needs of laboratories in developing nations. AB - To meet the biosafety requirements necessary to operate in the global scientific community, developing nations need the help of consultants from industrialized nations. PMID- 11381226 TI - Global harmonization of laboratory rodent health surveillance standards. AB - The authors discuss the need for international standards for the microbiologic and genetic quality of rodents. PMID- 11381225 TI - The international space station: globalizing life sciences. AB - The ISS changes the scope of science activities for the future and links the US to its partners in technology, science, and the exploration of space in an unprecedented manner. PMID- 11381227 TI - Mouse ranching 101. PMID- 11381228 TI - Newsfronts. PMID- 11381229 TI - Usda requests comments on definition and reporting of pain and distress. PMID- 11381230 TI - Ethicist or Bleeding Heart. PMID- 11381232 TI - Evaluation of a training program: a usda perspective. AB - Is your institution's animal care training program up to par? The author lists questions you'll need to answer. PMID- 11381231 TI - Healthy looking rabbit with a decreased appetite. PMID- 11381233 TI - Group housing female guinea pigs. AB - Group housing guinea pigs can save space and money, while improving housing standards. The authors describe enclosure design, enrichments, and husbandry techniques that facilitate group housing female guinea pigs. PMID- 11381234 TI - Technical team approaches to rodent care: cost savings, reduced risk, and improved stewardship. AB - With the increasing decentralization of animal care operations, lack of training and higher-density animal rooms make achieving humane assurances difficult. To offset these problems, some facilities have developed centralized animal care programs, including dispersed teams of qualified animal care specialists. PMID- 11381236 TI - Newsfronts. PMID- 11381235 TI - Respiratory protection for animal care workers. PMID- 11381237 TI - Phs proposes a new policy on instruction in the responsible conduct of research. PMID- 11381238 TI - Is a "best guess" good enough? PMID- 11381239 TI - Septicemia in la mancha goat kids. PMID- 11381240 TI - Developmental Neurobiology of Pain in Neonatal Rats. AB - The authors examine the biological basis for the behavioral evidence of pain perception in neonatal rats, and discuss the potential link to a greater understanding of chronic pain PMID- 11381241 TI - Water soluble propofol anesthesia: an effective and inexpensive alternative. AB - The authors discuss solubilized propofol, and demonstrate its acute use in guinea pigs as an effective, less expensive, easy-to-use alternative for laboratory animal anesthesia. PMID- 11381242 TI - Cardiac output measurement in mice by thermodilution. AB - Traditional methods for measuring cardiac output in mice are invasive and traumatic. The authors discuss using the less-invasive thermodilution method, which is widely accepted in humans and other animals. PMID- 11381243 TI - Selecting the right sterilization method for your surgical instruments. PMID- 11381244 TI - Stem cell decision time. PMID- 11381245 TI - Mitotic maneuvers in the light. PMID- 11381246 TI - Asking the age-old questions. PMID- 11381247 TI - Mitochondria: integrators in tumorigenesis? PMID- 11381248 TI - To be or not to BMP. PMID- 11381249 TI - Hypoxia and Lou Gehrig. PMID- 11381252 TI - Mouse models of cell death. AB - Cell death is critical for the development and orderly maintenance of cellular homeostasis in metazoans. Developmental genetics in model systems, including Caenorhabditis elegans and Drosophila melanogaster, have helped to identify and order the components of cell-death pathways. An even more complex network of apoptotic pathways has evolved in higher organisms that possess homologs within each set of cell-death regulators. Whereas biochemical studies provide details of molecular mechanisms, genetic models reveal the essential physiologic roles. Transgenic and gene-ablated mice have helped to elucidate mammalian apoptotic pathways and identify the principal effect of each cell death regulator. Here, we review the details of the apoptotic machinery as revealed by mice deficient in critical components of cell-death pathways; we concentrate on cell-death regulators classified as members of the caspase and Bcl2 families or, broadly, as adaptors and mitochondrial released factors. PMID- 11381253 TI - A conserved sorting-associated protein is mutant in chorea-acanthocytosis. AB - Chorea-acanthocytosis (CHAC, MIM 200150) is an autosomal recessive neurodegenerative disorder characterized by the gradual onset of hyperkinetic movements and abnormal erythrocyte morphology (acanthocytosis). Neurological findings closely resemble those observed in Huntington disease. We identified a gene in the CHAC critical region and found 16 different mutations in individuals with chorea-acanthocytosis. CHAC encodes an evolutionarily conserved protein that is probably involved in protein sorting. PMID- 11381254 TI - The gene encoding a newly discovered protein, chorein, is mutated in chorea acanthocytosis. AB - Chorea-acanthocytosis is a neurodegenerative disorder with peripheral red cell acanthocytosis. Linkage of chorea-acanthocytosis to chromosome 9q21 has been found. We refined the locus region and identified a previously unknown, full length cDNA encoding a presumably structural protein, which we called chorein. We found a deletion in the coding region of the cDNA leading to a frame shift resulting in the production of a truncated protein in both alleles of patients and in single alleles of obligate carriers. PMID- 11381255 TI - Mutations in the gene encoding lecithin retinol acyltransferase are associated with early-onset severe retinal dystrophy. AB - The chromophore of the visual pigments, 11-cis retinal, is derived from vitamin A (all-trans retinol) through a series of reactions that take place in retinal pigment epithelium (RPE); (ref. 1). The first of these reactions is catalyzed by lecithin retinol acyltransferase (LRAT); (ref. 2). We screened 267 retinal dystrophy patients for mutations in LRAT and identified disease-associated mutations (S175R and 396delAA) in three individuals with severe, early-onset disease. We showed that the S175R mutant has no acyltransferase activity in transfected COS-7 cells. Our findings highlight the importance of genetic defects in vitamin A metabolism as causes of retinal dystrophies and extend prospects for retinoid replacement therapy in this group of diseases. PMID- 11381256 TI - Mutations in the gene encoding c-Abl-binding protein SH3BP2 cause cherubism. AB - Cherubism (MIM 118400) is an autosomal dominant inherited syndrome characterized by excessive bone degradation of the upper and lower jaws followed by development of fibrous tissue masses, which causes a characteristic facial swelling. Here we describe seven mutations in the SH3-binding protein SH3BP2 (MIM 602104) on chromosome 4p16.3 that cause cherubism. PMID- 11381257 TI - Association analysis of NOTCH4 loci in schizophrenia using family and population based controls. AB - A genetic association between NOTCH4 and schizophrenia has previously been proposed. Unsing all markers previously shown to be associated, we found no evidence for such in three independent family-based samples (n=519 parent offspring trios), and a case-control sample derived from the same ethnic background as the original observation. These data strongly suggest that NOTCH4 is not a significant susceptibility allele for schizophrenia. PMID- 11381258 TI - Failure to confirm NOTCH4 association with schizophrenia in a large population based sample from Scotland. AB - The NOTCH4 gene was recently reported to be associated with schizophrenia based on TDT analysis of 80 British trios. The strongest evidence for association derived from two microsatellites. We genotyped both loci in a large sample of unrelated Scottish schizophrenics and controls, but failed to replicate the reported association, finding instead that each putative schizophrenia-associated allele had a somewhat lower frequency in schizophrenics than in controls. PMID- 11381259 TI - Deletion of the hypoxia-response element in the vascular endothelial growth factor promoter causes motor neuron degeneration. AB - Hypoxia stimulates angiogenesis through the binding of hypoxia-inducible factors to the hypoxia-response element in the vascular endothelial growth factor (Vegf) promotor. Here, we report that deletion of the hypoxia-response element in the Vegf promotor reduced hypoxic Vegf expression in the spinal cord and caused adult onset progressive motor neuron degeneration, reminiscent of amyotrophic lateral sclerosis. The neurodegeneration seemed to be due to reduced neural vascular perfusion. In addition, Vegf165 promoted survival of motor neurons during hypoxia through binding to Vegf receptor 2 and neuropilin 1. Acute ischemia is known to cause nonselective neuronal death. Our results indicate that chronic vascular insufficiency and, possibly, insufficient Vegf-dependent neuroprotection lead to the select degeneration of motor neurons. PMID- 11381260 TI - Regulation of the Caenorhabditis elegans longevity protein DAF-16 by insulin/IGF 1 and germline signaling. AB - The lifespan of Caenorhabditis elegans is regulated by the insulin/insulin-like growth factor (IGF)-1 receptor homolog DAF-2, which signals through a conserved phosphatidylinositol 3-kinase (PI 3-kinase)/Akt pathway. Mutants in this pathway remain youthful and active much longer than normal animals and can live more than twice as long. This lifespan extension requires DAF-16, a forkhead/winged-helix transcription factor. DAF-16 is thought to be the main target of the DAF-2 pathway. Insulin/IGF-1 signaling is thought to lead to phosphorylation of DAF-16 by AKT activity, which in turn shortens lifespan. Here, we show that the DAF-2 pathway prevents DAF-16 accumulation in nuclei. Disrupting Akt-consensus phosphorylation sites in DAF-16 causes nuclear accumulation in wild-type animals, but, surprisingly, has little effect on lifespan. Thus the DAF-2 pathway must have additional outputs. Lifespan in C. elegans can be extended by perturbing sensory neurons or germ cells. In both cases, lifespan extension requires DAF-16. We find that both sensory neurons and germline activity regulate DAF-16 accumulation in nuclei, but the nuclear localization patterns are different. Together these findings reveal unexpected complexity in the DAF-16-dependent pathways that regulate aging. PMID- 11381261 TI - High frequency of homoplasmic mitochondrial DNA mutations in human tumors can be explained without selection. AB - Researchers in several laboratories have reported a high frequency of homoplasmic mitochondrial DNA (mtDNA) mutations in human tumors. This observation has been interpreted to reflect a replicative advantage for mutated mtDNA copies, a growth advantage for a cell containing certain mtDNA mutations, and/or tumorigenic properties of mtDNA mutations. We consider another possibility-that the observed homoplasmy arose entirely by chance in tumor progenitor cells, without any physiological advantage or tumorigenic requirement. Through extensive computer modeling, we demonstrate that there is sufficient opportunity for a tumor progenitor cell to achieve homoplasmy through unbiased mtDNA replication and sorting during cell division. To test our model in vivo, we analyzed mtDNA homoplasmy in healthy human epithelial tissues and discovered that the model correctly predicts the considerable observed frequency of homoplasmic cells. Based on the available data on mitochondrial mutant fractions and cell division kinetics, we show that the predicted frequency of homoplasmy in tumor progenitor cells in the absence of selection is similar to the reported frequency of homoplasmic mutations in tumors. Although a role for other mechanisms is not excluded, random processes are sufficient to explain the incidence of homoplasmic mtDNA mutations in human tumors. PMID- 11381262 TI - Mutant glycosyltransferase and altered glycosylation of alpha-dystroglycan in the myodystrophy mouse. AB - Spontaneous and engineered mouse mutants have facilitated our understanding of the pathogenesis of muscular dystrophy and they provide models for the development of therapeutic approaches. The mouse myodystrophy (myd) mutation produces an autosomal recessive, neuromuscular phenotype. Homozygotes have an abnormal gait, show abnormal posturing when suspended by the tail and are smaller than littermate controls. Serum creatine kinase is elevated and muscle histology is typical of a progressive myopathy with focal areas of acute necrosis and clusters of regenerating fibers. Additional aspects of the phenotype include sensorineural deafness, reduced lifespan and decreased reproductive fitness. The myd mutation maps to mouse chromosome 8 at approximately 33 centimorgans (cM) (refs. 2, 4-7). Here we show that the gene mutated in myd encodes a glycosyltransferase, Large. The human homolog of this gene (LARGE) maps to chromosome 22q. In myd, an intragenic deletion of exons 4-7 causes a frameshift in the resultant mRNA and a premature termination codon before the first of the two catalytic domains. On immunoblots, a monoclonal antibody to alpha dystroglycan (a component of the dystrophin-associated glycoprotein complex) shows reduced binding in myd, which we attribute to altered glycosylation of this protein. We speculate that abnormal post-translational modification of alpha dystroglycan may contribute to the myd phenotype. PMID- 11381263 TI - Telomere dysfunction and evolution of intestinal carcinoma in mice and humans. AB - Telomerase activation is a common feature of advanced human cancers and facilitates the malignant transformation of cultured human cells and in mice. These experimental observations are in accord with the presence of robust telomerase activity in more advanced stages of human colorectal carcinogenesis. However, the occurrence of colon carcinomas in telomerase RNA (Terc)-null, p53 mutant mice has revealed complex interactions between telomere dynamics, checkpoint responses and carcinogenesis. We therefore sought to determine whether telomere dysfunction exerts differential effects on cancer initiation versus progression of mouse and human intestinal neoplasia. In successive generations of ApcMin Terc-/- mice, progressive telomere dysfunction led to an increase in initiated lesions (microscopic adenomas), yet a significant decline in the multiplicity and size of macroscopic adenomas. That telomere dysfunction also contributes to human colorectal carcinogenesis is supported by the appearance of anaphase bridges (a correlate of telomere dysfunction) at the adenoma-early carcinoma transition, a transition recognized for marked chromosomal instability. Together, these data are consistent with a model in which telomere dysfunction promotes the chromosomal instability that drives early carcinogenesis, while telomerase activation restores genomic stability to a level permissive for tumor progression. We propose that early and transient telomere dysfunction is a major mechanism underlying chromosomal instability of human cancer. PMID- 11381264 TI - Rapid gene mapping in Caenorhabditis elegans using a high density polymorphism map. AB - Single nucleotide polymorphisms (SNPs) are valuable genetic markers of human disease. They also comprise the highest potential density marker set available for mapping experimentally derived mutations in model organisms such as Caenorhabditis elegans. To facilitate the positional cloning of mutations we have identified polymorphisms in CB4856, an isolate from a Hawaiian island that shows a uniformly high density of polymorphisms compared with the reference Bristol N2 strain. Based on 5.4 Mbp of aligned sequences, we predicted 6,222 polymorphisms. Furthermore, 3,457 of these markers modify restriction enzyme recognition sites ('snip-SNPs') and are therefore easily detected as RFLPs. Of these, 493 were experimentally confirmed by restriction digest to produce a snip-SNP map of the worm genome. A mapping strategy using snip-SNPs and bulked segregant analysis (BSA) is outlined. CB4856 is crossed into a mutant strain, and exclusion of CB4856 alleles of a subset of snip-SNPs in mutant progeny is assessed with BSA. The proximity of a linked marker to the mutation is estimated by the relative proportion of each form of the biallelic marker in populations of wildtype and mutant genomes. The usefulness of this approach is illustrated by the rapid mapping of the dyf-5 gene. PMID- 11381265 TI - Deregulated expression of c-Myc depletes epidermal stem cells. AB - The beta-catenin/TCF signaling pathway is essential for the maintenance of epithelial stem cells in the small intestine. c-Myc a downstream target of beta catenin/TCF (ref. 2), can induce differentiation of epidermal stem cells in vitro. To determine the role of c-Myc in epidermal stem cells in vivo, we have targeted expression of human MYC2 to the hair follicles and the basal layer of mouse epidermis using a keratin 14 vector (K14.MYC2). Adult K14.MYC2 mice gradually lose their hair and develop spontaneous ulcerated lesions due to a severe impairment in wound healing; their keratinocytes show impaired migration in response to wounding. The expression of beta1 integrin, which is preferentially expressed in epidermal stem cells is unusually low in the epidermis of K14.MYC2 mice. Label-retaining analysis to identify epidermal stem cells reveals a 75% reduction in the number of stem cells in 3-month-old K14.MYC2 mice, compared with wildtype mice. We conclude that deregulated expression of c Myc in stem cells reduces beta1 integrin expression, which is essential to both keratinocyte migration and stem cell maintenance. PMID- 11381266 TI - Mitotic recombination is suppressed by chromosomal divergence in hybrids of distantly related mouse strains. AB - Mitotic recombination occurs with high frequency in humans and mice. It leads to loss of heterozygosity (LOH) at important gene loci and can cause disease. However, the genetic modulators of mitotic recombination are not well understood. As recombination depends on a high level of nucleotide sequence homology, we postulate that the frequency of somatic variants derived from mitotic recombination should be diminished in progeny from crosses between strains of mice in which nucleotide sequences have diverged. Here we report that mitotic recombination is suppressed, to various degrees in different tissues, in hybrids of distantly related mouse strains. Reintroduction of greater chromosomal homology by backcrossing restores mitotic recombination in offspring. Thus, chromosomal divergence inhibits mitotic recombination and, consequently, may act as a modifier of cancer susceptibility by limiting the rate of LOH. The suppression of mitotic recombination in some F1 hybrids in which meiotic recombination persists indicates that these processes are differentially affected by chromosomal divergence. PMID- 11381267 TI - Aberrant methylation of donor genome in cloned bovine embryos. AB - Despite recent successes in cloning various animal species, the use of somatic cells as the source of donor nuclei has raised many practically relevant questions such as increased abortion rates, high birth weight and perinatal death. These anomalies may be caused by incomplete epigenetic reprogramming of donor DNA. Genome-wide demethylation occurs during early development, 'erasing' gamete-specific methylation patterns inherited from the parents. This process may be a prerequisite for the formation of pluripotent stem cells that are important for the later development. Here, we provide evidence that cloned bovine embryos may have impaired epigenetic reprogramming capabilities. We found highly aberrant methylation patterns in various genomic regions of cloned embryos. Cloned blastocysts closely resembled donor cells in their overall genomic methylation status, which was very different from that of normal blastocysts produced in vitro or in vivo. We found demethylation of the Bov-B long interspersed nuclear element sequence in normal embryos, but not in cloned embryos, in which the donor type methylation was simply maintained during preimplantation development. There were also significant variations in the degree of methylation among individual cloned blastocysts. Our findings indicate that the developmental anomalies of cloned embryos could be due to incomplete epigenetic reprogramming of donor genomic DNA. PMID- 11381268 TI - A common polymorphism in the promoter of UCP2 is associated with decreased risk of obesity in middle-aged humans. AB - Obesity is the most common nutritional disorder in Western society. Uncoupling protein-2 (UCP2) is a recently identified member of the mitochondrial transporter superfamily that is expressed in many tissues, including adipose tissue. Like its close relatives UCP1 and UCP3, UCP2 uncouples proton entry in the mitochondrial matrix from ATP synthesis and is therefore a candidate gene for obesity. We show here that a common G/A polymorphism in the UCP2 promoter region is associated with enhanced adipose tissue mRNA expression in vivo and results in increased transcription of a reporter gene in the human adipocyte cell line PAZ-6. In analyzing 340 obese and 256 never-obese middle-aged subjects, we found a modest but significant reduction in obesity prevalence associated with the less-common allele. We confirmed this association in a population-based sample of 791 middle aged subjects from the same geographic area. Despite its modest effect, but because of its high frequency (approximately 63%), the more-common risk allele conferred a relatively large population-attributable risk accounting for 15% of the obesity in the population studied. PMID- 11381269 TI - Germline mutations of the gene encoding bone morphogenetic protein receptor 1A in juvenile polyposis. AB - Juvenile polyposis (JP; OMIM 174900) is an autosomal dominant gastrointestinal hamartomatous polyposis syndrome in which patients are at risk for developing gastrointestinal cancers. Previous studies have demonstrated a locus for JP mapping to 18q21.1 (ref. 3) and germline mutations in the homolog of the gene for mothers against decapentaplegic, Drosophila, (MADH4, also known as SMAD4) in several JP families. However, mutations in MADH4 are only present in a subset of JP cases, and although mutations in the gene for phosphatase and tensin homolog (PTEN) have been described in a few families, undefined genetic heterogeneity remains. Using a genome-wide screen in four JP kindreds without germline mutations in MADH4 or PTEN, we identified linkage with markers from chromosome 10q22-23 (maximum lod score of 4.74, straight theta=0.00). We found no recombinants using markers developed from the vicinity of the gene for bone morphogenetic protein receptor 1A (BMPR1A), a serine-threonine kinase type I receptor involved in bone morphogenetic protein (BMP) signaling. Genomic sequencing of BMPR1A in each of these JP kindreds disclosed germline nonsense mutations in all affected kindred members but not in normal control individuals. These findings indicate involvement of an additional gene in the transforming growth factor-beta (TGF-beta) superfamily in the genesis of JP, and document an unanticipated function for BMP in colonic epithelial growth control. PMID- 11381270 TI - Identification of the gene that, when mutated, causes the human obesity syndrome BBS4. AB - Bardet-Biedl syndrome (BBS, MIM 209900) is a heterogeneous autosomal recessive disorder characterized by obesity, pigmentary retinopathy, polydactyly, renal malformations, mental retardation, and hypogenitalism. The disorder is also associated with diabetes mellitus, hypertension, and congenital heart disease. Six distinct BBS loci map to 11q13 (BBS1), 16q21 (BBS2), 3p13-p12 (BBS3), 15q22.3 q23 (BBS4), 2q31 (BBS5), and 20p12 (BBS6). Although BBS is rare in the general population (<1/100,000), there is considerable interest in identifying the genes causing BBS because components of the phenotype, such as obesity and diabetes, are common. We and others have demonstrated that BBS6 is caused by mutations in the gene MKKS (refs. 12,13), mutation of which also causes McKusick-Kaufman syndrome (hydrometrocolpos, post-axial polydactyly, and congenital heart defects). MKKS has sequence homology to the alpha subunit of a prokaryotic chaperonin in the thermosome Thermoplasma acidophilum. We recently identified a novel gene that causes BBS2. The BBS2 protein has no significant similarity to other chaperonins or known proteins. Here we report the positional cloning and identification of mutations in BBS patients in a novel gene designated BBS4. PMID- 11381272 TI - Splitters and lumpers. PMID- 11381273 TI - An ongoing systematic update of hypertension recommendations. PMID- 11381274 TI - A matter of perspective. PMID- 11381275 TI - Message from the president. PMID- 11381276 TI - The 2000 Canadian Hypertension Recommendations: a summary. PMID- 11381277 TI - The 2000 Canadian recommendations for the management of hypertension: Part one- therapy. AB - OBJECTIVE: To provide updated, evidence-based recommendations for the therapy of hypertension in adults. OPTIONS: For patients with hypertension, there are a number of lifestyle manoeuvres and antihypertensive agents that may control blood pressure. Randomized trials evaluating first- line therapy with thiazides, beta adrenergic antagonists, angiotensin-converting enzyme inhibitors, calcium channel blockers, alpha-blockers, centrally acting agents or angiotensin II receptor antagonists were reviewed. OUTCOMES: The health outcomes considered were changes in blood pressure, cardiovascular morbidity, and cardiovascular and/or all-cause mortality rates. Economic outcomes were not considered due to insufficient evidence. EVIDENCE: Medline searches were conducted from the period of the last revision of the Canadian Recommendations for the Management of Hypertension (May 1998 to October 2000). Reference lists were scanned, experts were polled, and the personal files of the subgroup members and authors were used to identify other studies. All relevant articles were reviewed and appraised, using prespecified levels of evidence, by content experts and methodological experts. VALUES: A high value was placed on the avoidance of cardiovascular morbidity and mortality. BENEFITS, HARMS, AND COSTS: Various lifestyle manoeuvres and antihypertensive agents reduce the blood pressure of patients with sustained hypertension. In certain settings, and for specific classes of drugs, blood pressure lowering has been associated with reduced cardiovascular morbidity and/or mortality. RECOMMENDATIONS: The present document contains detailed recommendations pertaining to all aspects of the therapy of patients with hypertension, including lifestyle modifications proven to lower blood pressure, treatment thresholds, target blood pressures, choice of agents in various settings and strategies to enhance adherence. Lower thresholds for blood pressure treatment are advocated for people with other cardiovascular risk factors or established hypertensive target organ damage. Implicit in the recommendations for therapy is the principle that treatment should be individualized for each patient and the choice of agent should be dictated by coexistent conditions. For the treatment of uncomplicated essential hypertension, thiazides, beta-adrenergic antagonists, angiotensin converting enzyme inhibitors or calcium channel blockers may be appropriate, depending on individual circumstances. VALIDATION: All recommendations were graded according to strength of the evidence and voted on by the Canadian Hypertension Recommendations Working Group. Only those recommendations achieving high levels of consensus are reported here. These guidelines will be updated annually. PMID- 11381278 TI - Renal dysfunction after cardiac surgery. AB - OBJECTIVE: To assess the causes and outcomes of patients with postcardiac surgery renal dysfunction. PATIENTS AND METHODS: A large cardiovascular data- base including pre-, peri- and postoperative serum creatinine concentrations from 2214 consecutive cardiac surgery patients was analyzed. RESULTS: Sixty-nine patients developed postoperative renal dysfunction, defined as at least a 15 mL/min decline in the creatinine clearance rate resulting in a value of less than 40 mL/min. These patients were significantly older, and had a higher incidence of previous cardiac surgery, diabetes, obesity, peripheral vascular disease, hypertension and poor ventricular function. Postoperatively, these patients had a higher occurrence of low output syndrome and myocardial infarction. Stepwise logistic regression predictors of postoperative renal dysfunction included the following: postoperative low output syndrome; repeat cardiac surgery; being older than 65 years; having diabetes; having poor left ventricular function; and having had valve surgery. Preoperative renal dysfunction (defined as a creatinine clearance of less than 40 mL/min) was not found to be one of the predictors. The mean creatinine concentrations of patients with mild postoperative renal dysfunction (defined as a creatinine concentration of less than 200 mmol/L on the fourth or fifth postoperative day) decreased significantly at the fifth postoperative day, while that of patients with severe postoperative renal dysfunction rose to a mean of 300 mmol/L six months postoperatively. The incidence of late dialysis (defined as a need for dialysis after postoperative day 10) approached 30% among patients with severe postoperative renal dysfunction and only 2% among patients with mild postoperative renal dysfunction. The early mortality rate (during the first postoperative month) was similar in both groups and approached 30%. CONCLUSIONS: Patients who develop postoperative renal dysfunction have a high mortality rate. Postoperative low cardiac output is the most important cause of postoperative renal dysfunction and, therefore, should be avoided. Patients with creatinine concentrations of less than 200 mmol/L at postoperative day 4 or 5 will probably resume normal renal function. Patients with creatinine concentrations of more than 200 mmol/L at days 4 and 5 have a 30% chance of needing long term dialysis. PMID- 11381279 TI - Prognostic value of dipyridamole stress echocardiography in hypertensive patients with left ventricular hypertrophy, chest pain and resting electrocardiographic repolarization abnormalities. AB - BACKGROUND: Hypertension is a major cardiovascular risk factor in the development of coronary artery disease (CAD); therefore, evaluating the presence of CAD is a primary clinical goal. However, the noninvasive tests that are commonly used have poor diagnostic specificity, particularly in patients with left ventricular hypertrophy. OBJECTIVES: To assess the prognostic value of dipyridamole stress echocardiography (DET) for ischemic events in a subset of patients with hypertension with left ventricular hypertrophy, chest pain and resting electrocardiographic repolarization abnormalities. PATIENTS AND METHODS: Eighty two patients (48 men and 34 women; average age 65+/-7.2 years with left ventricular hypertrophy documented echocardiographically (left ventricular mass index greater than 50 g/h(2.7)), and resting ST segment shift of 0.1 mV or more from baseline at 80 ms after J point in at least two contiguous leads, were submitted to DET according to high-dosage protocol and coadministered with atropine. RESULTS: The follow-up period was 25.11+/-8.3 months. The stress test produced positive results in 30 patients (36.5%); 16 (53%) and three (5%) cardiac events occurred in positive and negative stress test groups, respectively. At multivariate analysis, only positive DET response (P=0.000002), left ventricular mass index (P=0.028) and a family history of CAD (P=0.037) were independent predictors. The two-year event-free survival rates were 95% and 47% (log-rank 21.093, P=0.00001) for negative and positive stress test results, respectively. CONCLUSIONS: DET is a useful tool in the prognostic assessment of coronary events in this particular subgroup of patients with hypertension. PMID- 11381281 TI - Ambulatory blood pressure monitoring in clinical practice. AB - Ambulatory blood pressure (ABP) monitoring has become increasingly more available in routine clinical practice in Canada. The ABP is more reliable and more reproducible than office readings, and is a better predictor of target organ damage. Normal values for ABP have been established using both cross-sectional and longitudinal outcome data. Abnormal mean 24 h and awake ABP values should exceed 135/85 mmHg and 140/90 mmHg, respectively. ABP recordings are useful in making a diagnosis of hypertension by identifying people with high office but normal ABP values. ABP monitoring can also be performed in patients already receiving antihypertensive therapy to determine the extent of any white coat effect that may be increasing office readings. The interpretation of the ABP should take into account cardiac risk factors, any target organ damage that may be present or coexisting conditions such as diabetes mellitus. PMID- 11381282 TI - Trapped renal arteries: functional renal artery stenosis due to occlusion of the aorta in the arch and below the kidneys. AB - Acute renal failure is a well recognized complication from the use of angiotensin converting enzyme inhibitors in patients with severe bilateral renovascular disease. A 54-year-old woman presented with acute pulmonary edema with intractable hypertension and a history of lower limb claudication. The addition of lisinopril to her antihypertensive regimen resulted, within 48 h, in the development of acute renal failure that remitted with cessation of the drug. She was found to have a heavily calcified occlusion of her aortic arch and another occlusion of the aorta below the renal arteries. Angiography and Doppler ultrasonography showed normal renal arteries. This is the first reported case of angiotensin-converting enzyme inhibitor-induced renal failure occurring in a patient with atherosclerotic occlusion of the aorta. The literature on suprarenal aortic occlusion is reviewed to determine the manner of presentation, prevalent risk factors and physical findings that typify this unique clinical entity. PMID- 11381283 TI - Acute myocardial ischemia associated with ingestion of bupropion and pseudoephedrine in a 21-year-old man. AB - A 21-year-old man presented to the emergency department with atypical chest pain, diaphoresis and shortness of breath. His electrocardiogram revealed ST segment elevation in leads II, III, aVF, V5 and V6, elevated creatine kinase-MB subunit levels and positive troponin I. He denied the use of cocaine, and smoking was his only risk factor for coronary artery disease. The patient was diagnosed with an acute myocardial infarction, yet an emergency coronary angiogram revealed normal coronary arteries. His medication history revealed recent commencement of bupropion for smoking cessation and pseudoephedrine as a nonprescription influenza remedy. It was postulated that this patient experienced acute coronary vasospasm in the presence of these two known sympathomimetic agents. The present case is the first report linking bupropion to an acute coronary syndrome, and one of a few cases associated with pseudoephedrine. PMID- 11381284 TI - Mixed aneurysmal and obstructive coronary artery disease causing acute myocardial infarction in a young woman with Takayasu's arteritis. AB - Takayasu's disease is a chronic inflammatory pathology of the aorta and its main branches. The present report describes a rare presentation of the disease in a young woman who presented with anterior myocardial infarction, and was subsequently found to have mixed aneurysmal and obstructive coronary artery disease involving the left anterior descending artery and its diagonal branches. Issues surrounding the management of coronary artery disease in this patient are discussed. PMID- 11381285 TI - The clinician scientist. PMID- 11381287 TI - Insulin-mediated blood flow and glucose uptake. AB - Normal aging is characterized by resistance to insulin-mediated vasodilation and glucose uptake. The mechanism or mechanisms responsible for resistance to the actions of insulin remain unclear. The majority of insulin-mediated glucose uptake occurs in skeletal muscle. It has recently been demonstrated that insulin increases skeletal muscle blood flow via an endothelium-derived, nitric oxide dependent mechanism. The authors' investigations of the relation between skeletal muscle blood flow and insulin-mediated glucose uptake in healthy elderly and young subjects are reviewed. PMID- 11381288 TI - The metabolism of bradykinin: a cornerstone for the understanding of the cardiovascular effects of metallopeptidase inhibitors. AB - The development of a sensitive and specific analytical approach for the quantification of bradykinin (BK) has enabled the identification of five metallopeptidases primarily responsible for the metabolism of BK, including angiotensin-converting enzyme (ACE) and neutral endopeptidase 24.11. Several metabolic studies have shown that the metallopeptidases that participate in BK metabolism vary by location (eg, plasma, endothelium or cardiomyocytes) and by pathology (eg, diabetes, myocardial infarction or left ventricular hypertrophy). These studies advance understanding of the role of endogenous BK in the cardiovascular effects of ACE inhibitors and other metallopeptidase inhibitors. PMID- 11381289 TI - Why were the results of the Heart Outcomes Prevention Evaluation (HOPE) trial so astounding? AB - The Heart Outcomes Prevention Evaluation (HOPE) study was important because it showed the benefits of ramipril - an angiotensin-converting enzyme (ACE) inhibitor - in patients at high risk for cardiovascular events. Treatment with ramipril significantly reduced the rates of death, myocardial infarction, stroke, coronary revascularization, cardiac arrest and heart failure, as well as the risk of diabetes-related complications and of diabetes itself. The effects of therapy with vitamin E were also evaluated, but no statistical benefits were shown. The benefits of ACE inhibitor therapy that were observed were likely due to a variety of mechanisms, not just a reduction in blood pressure. PMID- 11381290 TI - Use of measures of endothelial function to stratify risk. AB - Measures of endothelial dysfunction can be used to stratify risk in coronary artery disease and diabetes. Two studies are reviewed in which endothelial dysfunction was shown to be associated with an increased risk of cardiovascular events and to predict long term atherosclerotic disease. A study that showed reduced microvascular and macrovascular reactivity in subjects at risk for type 2 diabetes, and a study indicating that endothelial dysfunction precedes the development of microalbuminuria in type 1 diabetes are reviewed. PMID- 11381291 TI - Use of carotid ultrasound to stratify risk. AB - B-mode carotid ultrasound is a well validated imaging technique for evaluating the anatomical extent of subclinical atherosclerosis. It is used to measure arterial carotid intima-media thickness (IMT), and there is clear evidence that carotid IMT measurements can be used to identify individuals at increased risk for coronary events and stroke. The validity and the usefulness of the technique primarily as a research tool but also in the assessment of clinical cardiovascular risk - have been shown in a number of studies. PMID- 11381292 TI - Endothelial and myocyte apoptosis--role of angiotensin II. AB - Recent studies have assessed the role of angiotensin II in apoptosis and the effects of the angiotensin II type 1 (AT1) and type 2 (AT2) receptor subtypes. A complex picture is emerging with cell-specific effects of AT1 and AT2 receptors, and differential effects dependent on pathology. In vitro studies have shown that angiotensin II mediated apoptosis in myocytes and endothelium. More recent in vivo studies have shown the prevention and regression of vascular lesions by apoptosis induction with medications (including AT1 receptor inhibitor). PMID- 11381293 TI - Angiogenesis (clinical trials). AB - Therapeutic angiogenesis may play a role in the treatment of patients with vascular disease who have no other treatment option (those who are not candidates for bypass surgery or angioplasty). The first published clinical trial results appeared in 1998 and additional trials have since been completed or are ongoing. Results of several of these trials are reviewed. Studies of angiogenesis with basic fibroblast growth factor and human vascular endothelial growth factor suggest that these growth factors can induce functionally significant angiogenesis. Evolving delivery modalities and strategies are also highlighted. PMID- 11381294 TI - Case 1. Diabetes mellitus. AB - A sedentary 60-year-old man newly diagnosed with diabetes insists on seeing a cardiologist because he has read that diabetes damages blood vessels. Several questions submitted by participants and the corresponding answers (based on group discussion and on the actions taken for the real patient) are presented. PMID- 11381295 TI - Case 2. Hyperlipidemia. AB - A 58-year-old man with typical angina since 1994 presents to his physician. His angina is relatively stable and laboratory results indicate that his lipids are elevated. Recommendations for lipid management are discussed. PMID- 11381296 TI - High prevalence of celiac disease in patients with type 1 diabetes detected by antibodies to endomysium and tissue transglutaminase. AB - OBJECTIVE: To establish the prevalence of celiac disease (CD) in children with type 1 diabetes in British Columbia. PATIENTS AND METHODS: Two hundred thirty three children with type 1 diabetes were prospectively screened for CD using blind testing with the current 'gold standard', immunoglobulin A endomysium antibody (EmA), and the novel immunoglobulin A tissue transglutaminase (tTG) antibody. Those children with positive results were offered small bowel biopsy; a gluten-free diet was recommended if CD was confirmed. RESULTS: Nineteen children were positive for EmA and had an elevated tTG level. One patient from this group was already known to have CD, and the other 18 patients consented to biopsy. One biopsy was normal, three biopsies demonstrated elevated intraepithelial lymphocyte counts with normal morphology and 14 biopsies had morphological changes consistent with CD. Growth parameters were normal in all patients, and nine of 19 children who were positive for EmA were asymptomatic. Seven patients had mild elevation of tTG levels alone. Two children from this latter group had normal biopsies, and five declined biopsy. CONCLUSIONS: At least 14 new cases of CD were detected in addition to four known cases, yielding an overall biopsy confirmed prevalence of CD of 7.7% (18 of 233). The present study confirms that CD is as prevalent in the pediatric type 1 diabetic population in British Columbia as it is in Europe. Serological screening of these children is important because many children have no symptoms or signs suggestive of CD. This study suggests that tTG serology may also be useful in monitoring response and compliance with a gluten-free diet. PMID- 11381297 TI - Normal parathyroid function with decreased bone mineral density in treated celiac disease. AB - Decreased bone mineral density (BMD) has been reported in patients with celiac disease in association with secondary hyperparathyroidism. The present study investigated whether basal parathyroid hormone (PTH) remained elevated and whether abnormalities of parathyroid function were still present in celiac disease patients treated with a gluten-free diet. Basal seric measurements of calcium and phosphate homeostasis and BMD were obtained in 17 biopsy-proven patients under treatment for a mean period of 5.7+/-3.7 years (range 1.1 to 15.9). In addition, parathyroid function was studied with calcium chloride and sodium citrate infusions in seven patients. Basal measurements of patients were compared with those of 26 normal individuals, while parathyroid function results were compared with those of seven sex- and age-matched controls. Basal results were similar in patients and controls except for intact PTH (I-PTH) (3.77+/-0.88 pmol/L versus 2.28+/-0.63 pmol/L, P<0.001), which was higher in the former group but still within normal limits. Mean 25-hydroxy vitamin D and 1,25-dihydroxy vitamin D values were normal in patients. Parathyroid function results were also found to be similar in both groups. Compared with a reference population of the same age (Z score), patients had significantly lower BMDs of the hip (-0.60+/ 0.96 SDs, P<0.05) and lumbar spine (-0.76+/-1.15 SDs, P<0.05). T scores were also decreased for the hip (-1.3+/-0.9 SDs, P<0.0001) and lumbar spine (-1.4+/-1.35 SDs, P<0.0001), with two to three patients being osteoporotic (T score less than 2.5 SDs) and seven to eight osteopenic (T score less than -1 SDs but greater than or equal to -2.5 SDs) in at least one site. Height and weight were the only important determinants of BMD values by multivariate or logistical regression analysis in these patients. The results show higher basal I-PTH values with normal parathyroid function in treated celiac disease. Height and weight values are, but I-PTH values are not, an important determinant of the actual bone mass of patients. Normal parathyroid function in treated patients suggests a lack of previous severe secondary hyperparathyroidism and/or complete adaptation to prior changes in parathyroid function. PMID- 11381298 TI - Meckel's diverticulum in Crohn's disease. AB - Meckel's diverticulum is a congenital abnormality of the distal ileum associated with failed vitelline duct closure. Detailed pathological studies have estimated its frequency to be about 2% of the general population, and it has been anecdotally recorded in patients with Crohn's disease. Most patients with Crohn's disease have imaging studies of the small intestine during the course of their disease, and often, an intestinal resection. Thus, it seems possible to estimate the prevalence of Meckel's diverticula in Crohn's disease. In addition, patient characteristics may be important, especially if management of Crohn's disease is altered. Of 877 patients with Crohn's disease, 10 (about 1%) had a Meckel's diverticulum diagnosed, including six men and four women. All were diagnosed with Crohn's disease before age 50 years and seven were diagnosed before age 30 years. There were five with ileocolonic disease, two with colon-only disease and three with ileum-only disease. The clinical behaviour of five patients could be classified as penetrating and two as stricturing. A total of 311 patients had an ileocolonic resection, including eight (about 2%) with a Meckel's diverticulum. In contrast to some case reports, no heterotopic mucosa was detected and the Meckel's diverticulum was incidental and, apparently, an unexpected finding. In each case, the diverticulum was not involved with Crohn's disease but was included in the ileal resection. These results suggest that the overall prevalence of a Meckel's diverticulum is not increased in Crohn's disease but may result in resection of additional small intestine. PMID- 11381299 TI - Bioartificial liver support. AB - Bioartificial liver support has been increasingly the focus of both basic and clinical research in an attempt to replicate the multiplicity of normal liver function. The concept is attractive because, if it is effective, patients with acute liver failure may be supported until native liver regeneration occurs or, by optimizing their condition, until liver transplantation is possible. Current bioartificial liver support systems utilize primary porcine hepatocytes or transformed human hepatocytes, which are housed within a bioreactor, through which the patient's blood or plasma is pumped in an extracorporeal circuit. The optimal source for the hepatocytes is an area of debate; however, a genetically engineered cell line may provide optimal function. Novel three-dimensional matrices that anchor the hepatocytes are being designed to mimic architectural features of the normal liver. Large multicentre, randomized, controlled trials are ongoing following several pilot studies. Serious side effects such as hemodynamic instability and immune reactions have been infrequent. Much controversy, however, surrounds the issue of possible transmission of pig endogenous retrovirus to humans, and current trials are being carefully monitored. Bioartificial liver support is a promising technology, and the results of current and planned studies are awaited with great interest. PMID- 11381300 TI - The clinical risks of infection associated with endoscopy. AB - The cleaning of flexible endoscopes is difficult and time consuming. Any method of attempted sterilization or high level disinfection will fail if prior cleaning has been defective. Inadequate reprocessing of endoscopes may result in patient to patient transmission of serious bacterial and viral diseases or infection with endemic hospital pathogens. Antibiotic prophylaxis is required to prevent septicemia and bacterial endocarditis in high risk patients undergoing specific endoscopic procedures. Prevention of serious endoscopy-associated clinical infections requires strict compliance with detailed reprocessing protocols by specially trained nursing staff. PMID- 11381302 TI - Cecal carcinoma: initially diagnosed as Crohn's disease on small bowel follow through. AB - Lesions in the terminal ileum are often difficult to visualize on routine small bowel follow-through (SBFT) and may require further investigation to rule out associated abnormalities in the ileocecal valve or cecum. This may be done by peroral pneumocolon at the same sitting as the SBFT, but may require bowel preparation. Two cases of cecal carcinoma that were initially diagnosed as Crohn's disease on SBFT without further investigation of the cecum are reported. PMID- 11381301 TI - Acute hepatitis induced by alpha-interferon in a patient with chronic hepatitis C. AB - Hepatic adverse effects occur very rarely with alpha-interferon therapy. A case of acute hepatitis induced by alpha-interferon in a 33-year-old man with chronic hepatitis C is described. The patient developed acute hepatitis with very high aminotransferase activity and jaundice. After discontinuing alpha-interferon therapy, hepatitis resolved rapidly. The immune-mediated mechanism is the most probable cause of this hepatitis. PMID- 11381303 TI - Familial microscopic colitis. AB - Collagenous and lymphocytic colitis are two inflammatory conditions of the colon that are often collectively referred to as microscopic colitis. The present report describes what is believed to be the third published case of familial microscopic colitis. A 55-year-old woman who suffered from chronic diarrhea was diagnosed with lymphocytic colitis on colonic biopsy. Subsequently, her 36-year old daughter was diagnosed with collagenous colitis. The familial occurrence of these diseases may support an immunological hypothesis for their etiology. In addition, it supports the assumption that collagenous and lymphocytic colitis are two manifestations of the same disease process rather than two completely separate entities. The familial tendency of this disease may make a case for early colonoscopy and biopsy in relatives of patients diagnosed with microscopic colitis if they present with suggestive symptoms. PMID- 11381304 TI - Role of reference materials used in measurement of oil pollution and a correlation equation to determine oil in seawater. PMID- 11381305 TI - Fish cell line as an in vitro test system for analyzing chromosome aberrations. PMID- 11381306 TI - Cytotoxic effects of beta-thujaplicin on rat thymocytes and prevention by the compound in tributyltin-induced thymocyte damage. PMID- 11381307 TI - Determination of safe work time and exposure control need for pesticide applicators. PMID- 11381308 TI - Larval growth in postlarvae of Penaeus indicus on exposure to lead. PMID- 11381309 TI - Exposure to natural radioactivity from thermal waters in Croatia. PMID- 11381310 TI - Organochlorine residues in human blood from Nainital (U.P.), India. PMID- 11381311 TI - Separation of inorganic arsenic species in groundwater using ion exchange method. PMID- 11381312 TI - Bacterial luminescence test screening of highly polluted areas in the Odra River. PMID- 11381313 TI - Mercury fractions in natural and urban soils of the Middle Amur, Far East Russia. PMID- 11381314 TI - Enhanced accumulation of lead in Brassica pekinensis by soil-applied chloride salts. PMID- 11381315 TI - Metal (Cd, Pb, Cu, Zn, Fe, Cr, Ni) concentrations in tissues of a fish Sardina pilchardus and a prawn Peaenus japonicus from three stations on the Mediterranean Sea. PMID- 11381316 TI - Copper and zinc uptake by spring wheat (Triticum aestivum L.) and corn (Zea mays L.) grown in Baiyin region. PMID- 11381317 TI - Bioaccumulation of arsenic in aquacultural large-scale mullet Liza macrolepis from blackfoot disease area in taiwan. PMID- 11381318 TI - Bioremediation of methylparathion by free and immobilized cells of Bacillus sp. isolated from soil. PMID- 11381319 TI - Assessment of the ready biodegradability of Bisphenol A. PMID- 11381320 TI - Formulation and tillage effects on atrazine and alachlor in shallow ground water in upland corn production. PMID- 11381321 TI - Lindane, chlorpyriphos, and quinalphos residues in mustard seed and oil. PMID- 11381322 TI - Comparison of organochlorine pesticide levels in soil and groundwater of Agra, India. PMID- 11381323 TI - Foliar biochemical features of plants as indicators of air pollution. PMID- 11381324 TI - Polycyclic aromatic hydrocarbons and polychlorinated biphenyl contamination in DaTong City, China. PMID- 11381325 TI - Study on indoor air pollutants: toxicity screening of suspended particulate matter. PMID- 11381326 TI - Production of beta-mannanase and beta-mannosidase from Aspergillus awamori K4 and their properties. AB - beta-Mannanase and beta-mannosidase from Aspergillus awamori K4 was produced by solid culture with coffee waste and wheat bran. The optimum composition for enzyme production was 40% coffee waste-60% wheat bran. Two enzymes were partially purified. Optimum pH was about 5 for both enzymes, and optimum temperature was around 80 degrees C for beta-mannanase and 60-70 degrees C for beta-mannosidase. These enzymes produced some oligosaccharides from glucomannan and galactomannan by their hydrolyzing and transferring activities. beta-Mannanase hydrolyzed konjak and locust bean gum 39.1% and 15.8%, respectively. Oligosaccharides of various molecular size were released from glucomannan of konjak, but on the addition of cellulase, mannobiose was released selectively. In locust bean gum, tetra-, tri-, and disaccharides (mannobiose) were mainly released by K4 beta mannanase. Tetra- and trisaccharides were heterooligosaccharides consisting of galactose and mannose residues. K4 beta-mannosidase had a transglycosylation action, transferring mannose residue to alcohols and sugars like fructose. PMID- 11381327 TI - Adherence of Shigella dysenteriae 1 to human colonic mucin. AB - The pathogenic potential of Shigella is correlated with the ability of the organism to invade and multiply within the cells of colonic epithelium. Although invasion is the ultimate event, a preceding step is adherence. Shigella dysenteriae 1 preferentially adhered to colonic mucin and not to small intestinal mucin. The pathogen showed a very strong adherence pattern to human colonic mucin when compared with guinea pig and rat mucin. The adherence pattern of S. dysenteriae 1 was not altered on preincubation with monosaccharides present in mucins, suggesting that the receptor for the pathogen is not a simple sugar. Binding of S. dysenteriae 1 to human colonic mucin was not by weak hydrophobic forces. The bacterium also adhered to glycolipids, emphasizing the role of glycoconjugates as receptors for S. dysenteriae 1. PMID- 11381328 TI - Cellular localization and characterization of the Bacillus thuringiensis Orf2 crystallization factor. AB - In vivo crystallization of the Bacillus thuringiensis insecticidal protein toxin Cry2Aa has been studied in the presence and absence of its associated crystallization factor Orf2. When expressed in the same cell, both proteins were localized to the cuboidal cytoplasmic crystal. When expressed in the absence of Cry2Aa, Orf2 was randomly distributed throughout the cytoplasm. Cry2Aa when expressed in the absence of Orf2 did not form visible crystals but could be recovered from the insoluble fraction of the cell lysate and solubilized at high pH. Purified Orf2 was found to be soluble over a wide range of pH although it could be co-precipitated in the presence of Cry2Aa, suggesting a direct interaction between the two. PMID- 11381329 TI - Phototrophic N2 fixation suppressed by activated sulfate reduction in anoxic rice soil slurries. AB - Interactions between sulfate reduction (SR) and phototrophic nitrogenase activities were investigated in rice soil slurries mixed with rice straw. Activation of SR by adding exogenous sulfate suppressed acetylene-reducing activity (ARA) of the slurries, which was associated with phototrophic purple bacteria (PB) enumerated to 108-109 MPN g-1 dry weight (dw) soil. Adding 5 mm sodium molybdate, an inhibitor of SR, markedly increased ARA. However, in the slurries receiving both molybdate and exogenous sulfate, the effects declined simultaneously with partial recovery of SR. These results indicate outcompetition of sulfate-reducing bacteria (SRB) with PB in rice soil, when sulfate concentrations are high enough to support SR. The increasing effects of molybdate on ARA continued during the incubation in the sulfate-depleted condition, probably because of absence of SR and toxicity of molybdate to methanogenesis. Accordingly, stopping activities of the competitive microorganisms may be efficient to increase N2 fixation in rice soil. PMID- 11381330 TI - Toxicity and mutagenesis of chrysotile asbestos to Agrobacterium radiobacter. AB - The mutation of Agrobacterium radiobacter cells exposed to chrysotile asbestos was examined by the random amplified polymorphic DNA (RAPD) method. Approximately 1.4 kbp of DNA in A. radiobacter, which was not amplified strongly in the cells that were not exposed to asbestos, was amplified in the cells that were exposed to asbestos. Mutation in genomic DNA of A. radiobacter was found to be induced by asbestos. Specific DNA that was amplified by asbestos present in PCR products and that which exists latently in genomic DNA were cloned, and these sequences were then determined and compared. It was shown that one of the mutations by the asbestos in the A. radiobacter occurred only in the primer annealed region and was a point mutation or deletion. PMID- 11381331 TI - Molecular cloning of cgrA, the gene encoding the Aspergillus nidulans ortholog of Saccharomyces cerevisiae CGR1. AB - Saccharomyces cerevisiae CGR1 encodes a 120-amino acid protein with a predominant nucleolar localization. In this study we report the identification and cloning of the ortholog, cgrA, from Aspergillus nidulans. The cgrA gene is comprised of three exons on A. nidulans Chromosome 7. The cDNA contains a single open reading frame (ORF) that would encode a protein of 114 amino acids with 44% sequence identity to yeast Cgr1p. A plasmid expressing cgrA complemented the impaired growth phenotype of a yeast strain that can be inducibly depleted of CGR1, and a green fluorescent protein (GFP)-tagged CgrA protein had the same nucleolar localization as the corresponding yeast protein. These results identify cgrA as the A. nidulans ortholog of yeast CGR1 and suggest evolutionary conservation of nucleolar localization mechanisms used by these proteins. PMID- 11381332 TI - Photoinactivation of Acinetobacter baumannii and Escherichia coli B by a cationic hydrophilic porphyrin at various light wavelengths. AB - Photodynamic treatment by the cationic TMPyP photosensitizer was undertaken on the multiple antibiotic-resistant bacteria Acinetobacter baumannii and Escherichia coli. Total eradication of the bacterial cultures was determined immediately after initiation of illumination when these bacteria were treated with 5, 10, 15, 20-tetra (4-N methylpyridyl)porphine (TMPyP) at a concentration of 29.4 micromol/L and illuminated by blue, green, or red light. Total eradication of both bacteria was obtained also after treatment of bacterial cultures with 3.7 micromol/L TMPyP and illumination with blue light (400-450 nm). On the other hand, an 8- or 16- to 20-fold higher light intensity, respectively, was required for total eradication upon illumination with green (480-550 nm) or red light (600-700 nm). A 407-nm blue light only 7 and 9 joules/cm2, respectively, was needed for total eradication of both bacteria even at a concentration of 3.7 micromol/L TMPyP. X-ray-linked microanalysis demonstrated loss of potassium and a flood of sodium and chloride into the cells, indicating serious damage to the cytoplasmic membrane. Transmission electron microscopy (TEM) revealed structural changes and damage to the membrane of treated E. coli. In A. baumannii-treated cells, mesosomes and black dots that resemble aggregation of polyphosphate polymers could be seen. DNA breakage appeared only after a long period of illumination, when the bacterial cell was no longer viable. It can be concluded that cytoplasmic membrane damage and not DNA breakage is the major cause for bacterial death upon photosensitization. PMID- 11381333 TI - A simple and efficient method for the purification of membrane-bound levansucrase from Zymomonas mobilis. AB - A new and efficient method for the purification of levansucrase from cell-free extracts of a flocculant mutant of Zymomonas mobilis ATCC 10988 was developed. Levansucrase activity was almost completely recovered and purified by a factor of 15 after precipitation with 0.1 m MnCl2 as a first capturing step. The enzyme was homogeneously purified by ultrafiltration and anion-exchange chromatography and exhibited a levan-forming activity of 39.2 U mg-1. The native enzyme formed large aggregates with an apparent molecular mass of more than 106 Da as determined by size-exclusion chromatography, whereas denaturing SDS-PAGE indicated an apparent molecular mass of 50 kDa for the subunits. PMID- 11381334 TI - Phylogenetic analysis of vertically transmitted psyllid endosymbionts (Candidatus Carsonella ruddii) based on atpAGD and rpoC: comparisons with 16S-23S rDNA derived phylogeny. AB - Psyllids are insects that harbor endosymbionts (Candidatuus Carsonella ruddii) within specialized cells found in the insect's body cavity. Previous phylogenetic analyses based on endosymbiont 16S-23S ribosomal DNA and a host gene were concordant (M.L. Thao, et al., Appl. Env. Microbiol. 66:2898, 2000). Additional analyses with atpAGD and rpoBC gave similar trees showing the agreement expected from organisms that evolve through vertical transmission with no gene exchange. PMID- 11381335 TI - Bacillus thuringiensis: a common member of microflora in activated sludges of a sewage treatment plant. AB - Bacillus thuringiensis was recovered at a high frequency from activated-sludge system environments in an urban sewage-digestive plant. All of the test materials, sampled at several digesting steps, contained the organism. Of 515 colonies belonging to the B. cereus/B. thuringiensis group, 45 (8.7%) were assigned to B. thuringiensis. The highest density of this bacterium was 1.6 x 103 cfu/ml in a scum sample of the first aeration basin. Among the 45 isolates, 7 were assigned to the known H serovars. Two isolates of the serovar kenyae isolates exhibited Lepidoptera-specific toxicity. Diptera-specific toxicity was shown by an isolate of serovar israelensis and a serologically undefined isolate. Lectin activity was associated with 12 isolates. PMID- 11381336 TI - Identification of essential cysteine residues in 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase from Corynebacterium glutamicum. AB - To ascertain the functional role of cysteine residue in 3-deoxy-d-arabino heptulosonate-7-phosphate (DAHP) synthase from Corynebacterium glutamicum, site directed mutagenesis was performed to change each of the three residues to serine. Plasmids were constructed for high-level overproduction and one-step purification of histidine-tagged DAHP synthase. Analysis of the purified wild type and mutant enzymes by SDS-polyacrylamide gel electrophoresis showed an apparent protein band with a molecular mass of approximately 45 kDa. Cys145Ser mutant retained about 16% of the enzyme activity, while DAHP synthase activity was abolished in Cys67Ser mutant. Kinetic analysis of Cys145Ser mutant with PEP as a substrate revealed a marked increase in Km with significant change in kcat, resulting in a 13.6-fold decrease in kcat/KmPEP. Cys334 was found to be nonessential for catalytic activity, although it is highly conserved in DAHP synthases. From these studies, Cys67 appears important for synthase activity, while Cys145 plays a crucial role in the catalytic efficiency through affecting the mode of substrate binding. PMID- 11381337 TI - Characterization of Methanococcus voltaei strain P2F9701a: a new methanogen isolated from estuarine environment. AB - A new methanogen, designated as strain P2F9701a (= OCM 745), was isolated from a water sample of estuarine environment in Elrin Shi, Taiwan. Cells of strain P2F9701a were motile coccus (0.7 approximately 1.1 micron) with tufts of flagella. Gas vacuoles were observed, and the protein cell wall was composed of S layer protein subunit with Mr of 74,700. Cells catabolized formate and H2+CO2 to produce methane, but not acetate, methanol, and trimethylamine. Strain P2F9701a grew in the range of 30-42 degrees C, with optimal growth temperature at 37 degrees C, but did not grow below 28 degrees C or above 42 degrees C. This estuarine isolate P2F9701a tolerated well the NaCl concentration between 0.02 and 1.03 m, and the optimal salt for growth was 0.17 m. Although phylogenetic analytic results indicated that P2F9701a belongs to the mesophilic, hydrogenotrophic marine methanogen of Methanococcus voltaei, the occurrence of gas vacuoles, tufts of flagella, eury-halotolerant and steno-thermotolerant characters of strain P2F9701a are different from mesophilic Methanococcus spp. that had been reported. PMID- 11381338 TI - Frankia KB5 possesses a hydrogenase immunologically related to membrane-bound. AB - The immunological relationship of the hydrogenase in Frankia KB5 to hydrogenases in other microorganisms was investigated using antisera raised against holo [NiFe]-hydrogenases isolated from Alcaligenes latus, Azotobacter vinelandii, Ralstonia eutropha, and the small and large hydrogenase subunits from Bradyrhizobium japonicum. The antisera raised against the A. latus, R. eutropha, and B. japonicum (large subunit) polypeptides were found to recognize two polypeptides, corresponding to the unprocessed and processed forms of the hydrogenase subunit in Frankia KB5. None of the antisera, including the antibodies produced against the small hydrogenase subunit isolated from B. japonicum, recognized any polypeptide related to the small hydrogenase subunit in Frankia KB5. An immunogold localization study of the intracellular distribution of hydrogenase in Frankia KB5, with the cryo-section technique, showed that labeling in the membrane of both hyphae and vesicles was positively correlated with hydrogenase activity. PMID- 11381339 TI - Oxidation of L-thiazolidine-4-carboxylate by delta1-pyrroline-5-carboxylate reductase in Escherichia coli. AB - L-Thiazolidine-4-carboxylate (T4C, thiaproline) is a sulfur-containing proline analog that stimulates the immune system in aging mice and inhibits urinary tract pathogens such as Escherichia coli. A constitutive NADP+-dependent T4C dehydrogenase activity was detected in the soluble fraction of a putA::Tn5 mutant of E. coli lacking l-proline dehydrogenase and partially purified by ammonium sulfate precipitation, dye-affinity chromatography on Cibacron Blue 3GA agarose, and ion-exchange chromatography on DEAE-cellulose. At each step in the purification, T4C dehydrogenase activity copurified with Delta1-pyrroline-5 carboxylate (P5C) reductase activity. E. coli strains with greatly reduced P5C reductase activity due to a proC mutation had no detectable T4C dehydrogenase activity. Although P5C reductase did not act on proline, it also catalyzed the oxidation of 3,4-dehydroproline. These results suggest that this biosynthetic enzyme may play a role in the degradation of proline analogs and limit the clinical efficacy of these compounds. PMID- 11381340 TI - Patch testing with 20% Dermatophagoides pteronyssinus/farinae (Chemotechnique) antigen. AB - BACKGROUND: Patch testing with dust mite antigens might identify mite-sensitive individuals, particularly those with atopic dermatitis who can benefit from avoidance measures. Currently available dust mite allergens have not been well studied. OBJECTIVE: To determine the proper dilution of 20% Dermatophagoides pteronyssinus/farinae mix antigen (Chemotechnique, Malmo, Sweden) for use in closed patch testing. METHODS: Eighteen nonatopic, healthy control subjects were patch-tested to the 20% concentration, yielding 15 (83%) positive reactions, most showing a decrescendo or persistent pattern suggesting an inordinately high number of false positive reactions. Dilutions of 1.25% to 0.1% in white petrolatum were used in patch testing 8 atopic dermatitis and 11 respiratory atopy patients, and 12 nonatopic controls. RESULTS: Positive reaction rates to the 0.25% and 0.1% concentrations, respectively, were 87.5% and 62.5% for atopic dermatitis, 54% and 18% for respiratory atopy, and 33% and 8% for healthy controls. Using Fisher's exact test, the 0.1% dilution was shown to significantly differentiate rates of positivity among the 3 groups, particularly between atopic dermatitis subjects and healthy controls. CONCLUSION: We find that a 0.1% dilution of 20% D. pteronyssinus/farinae mix antigen (Chemotechnique) to be useful in identifying mite-allergic individuals with atopic dermatitis. PMID- 11381341 TI - A preliminary report of the occupation of patients evaluated in patch test clinics. AB - BACKGROUND: The interplay between the occupational environment and worker's skin can result in contact dermatitis of both irritant and allergic types. Other forms of dermatitis can also be influenced by occupational exposures. OBJECTIVE: The aim of this study is to compare the occupations and allergens of occupational contact dermatitis cases with nonoccupational contact dermatitis cases. METHODS: Diagnostic patch testing with allergens of the North American Contact Dermatitis Group and occupational coding by the National Institute for Occupational Safety and Health methods. RESULTS: Of 2,889 patients referred for evaluation of contact dermatitis, 839 patients (29%) were found to have occupational contact dermatitis. Of the 839 cases deemed occupational, 455 cases (54%) were primarily allergic in nature and 270 cases (32%) were primarily irritant in nature. The remaining 14% were diagnoses other than contact dermatitis, aggravated by work. The occupation most commonly found to have allergic contact dermatitis was nursing. Allergens strongly associated with occupational exposure were thiuram, carbamates, epoxy, and ethylenediamine. CONCLUSION: Some contact allergens are more commonly associated with occupational contact dermatitis. Nursing and nursing support are occupations most likely to be overrepresented in contact dermatitis clinics. PMID- 11381342 TI - Comparison of patch test results with a standard series among white and black racial groups. AB - BACKGROUND: Environmental, cultural, occupational, genetic, individual, and racial differences are important factors in the study of contact dermatitis. Some epidemiologic studies have compared overall sensitization rates among different racial groups, but similar data are lacking on individual allergens. OBJECTIVE: Determine differences in sensitization rates between 2 racial groups in North America undergoing patch testing over a period of 4 years at the Cleveland Clinic Foundation (CCF), Ohio. METHODS: Retrospective computer review of the standard screening tray results of 991 patients with an average age of 45.9 years consisting of 877 (88.5%) whites and 114 (11.5%) blacks. RESULTS: Nickel sulfate and thiomersal (both 8.0%) and nickel sulfate and p-phenylenediamine (both 10.6%) were the 2 most common sensitizers among whites and blacks respectively. There was a statistically significant difference (P =.00599) in the sensitization rate for p-phenylenediamine in blacks (10.6%) compared with whites (4.5%). There were also statistically significant differences in sensitization rates for p phenylenediamine (21.2%; P =.00005) and imidazolidinyl urea in petrolatum (pet.) (9.1%; P =.04103) in black men compared with white men (p-phenylenediamine [4.2%] and imidazolidinyl urea [2.6% pet.]). CONCLUSION: The differences in sensitization rates, especially for p-phenylenediamine, may reflect variations in allergen exposure among racial groups or interindividual variations in the N acetylation (N-acetyltransferase 1 [NAT1] and 2 [NAT2]) capacities of human skin for p-phenylenediamine. PMID- 11381344 TI - Results of evaluating health care workers with prick and patch testing. AB - BACKGROUND: Health care workers are exposed to many agents that can cause irritant or allergic contact dermatitis. Recently, much attention has been focused on latex sensitivity, which commonly causes contact urticaria. Most studies have examined the conditions of irritant or allergic contact dermatitis and contact urticaria independently. Therefore, we have little information about the possible occurrence of these conditions in the context of combined assessment including both prick and patch testing. OBJECTIVE: To determine the prevalence of irritant and allergic contact dermatitis and contact urticaria in a group of health care workers presenting with skin problems. METHODS: Retrospective review of health care workers assessed by both prick and patch testing in an occupational health clinic. RESULTS: The diagnoses included 61% with irritant contact dermatitis, 31% with allergic contact dermatitis, and 27% with contact urticaria to latex. Eleven percent had both allergic contact dermatitis related to thiuram and contact urticaria to latex. Ninety five percent were deemed to be work-related. CONCLUSION: Health care workers presenting with skin complaints should be assessed with both prick and patch testing. PMID- 11381343 TI - A multicenter study of patch test reactions with dental screening series. AB - BACKGROUND: Dental products contain many allergens, and may cause problems both for patients undergoing dental treatment and for dental personnel because of occupational exposure. Individual patch test clinics may not study sufficient numbers of patients to collect reliable data on uncommon allergens. OBJECTIVE: To collect information on dental allergens based on a multicenter study. MATERIALS AND METHODS: The Finnish Contact Dermatitis Group tested more than 4,000 patients (for most allergens, 2,300 to 2,600 patients) with dental screening series. Conventional patch testing was performed. The total number and percentage of irritant (scored as irritant [IR] or doubtful [?]) and allergic (scored as +, ++, or +++) patch test reactions, respectively, were calculated, as well as the highest and lowest percentage of allergic patch test reactions recorded by the different patch test clinics. A reaction index (RI) was calculated, giving information on the irritancy of the patch test substances. RESULTS: The most frequent allergic patch test reactions were caused by nickel (14.6%), ammoniated mercury (13%), mercury (10.3%), gold (7.7%), benzoic acid (4.3%), palladium (4.2%) and cobalt (4.1%). 2-hydroxyethyl methacrylate (2.8%) provoked most of the reactions caused by (meth)acrylates. Menthol, peppermint oil, ammonium tetrachloroplatinate, and amalgam alloying metals provoked no (neither allergic nor irritant) patch test reactions. CONCLUSION: Patch testing with allergens in the dental screening series, including (meth)acrylates and mercury, needs to be performed to detect contact allergy to dental products. PMID- 11381345 TI - Contact allergy to balsam of Peru. II. Patch test results in 102 patients with selected balsam of Peru constituents. AB - BACKGROUND: In Switzerland, Germany, and Austria, allergic reactions to balsam of Peru (BP) have now made it the third most common contact allergen. OBJECTIVE: A series of 20 single BP constituents (including resorcinol monobenzoate), established in 1995, was used for patch tests in patients with a positive reaction to BP in the standard series. MATERIALS AND METHODS: Between 1995 and 1998, 2,273 patients were tested with the standard series, including BP, fragrance mix (FM), and propolis. Patients positive for BP were requested to participate in a further test using the 19 compounds of the BP constituents and resorcin monobenzoate (BP series); 102 patients agreed and were patch tested. The results of the 72-hour reading were used for the evaluation. RESULTS: A total of 93 patients reacted to 1 or more of the BP series compounds. Positive reactions were seen, in decreasing order, to cinnamic alcohol, cinnamic acid, coniferyl benzoate, benzoic acid, cinnamyl cinnamate, eugenol, resorcinol monobenzoate, coniferyl alcohol, and benzyl alcohol. There were no positive reactions to vanillin or ferulic acid. A correlation between skin lesions and frequent consumption of sweets was found in 7 patients with major positive test reactions to coniferyl benzoate and benzyl alcohol. Most of the reactions to eugenol and isoeugenol had less to do with BP itself than with a primary sensitization to fragrances. Although resorcin monobenzoate (RMB) has up to now not been detected in BP, 16 patients reacted distinctly to this compound. Eleven were strong smokers; the remaining ones had contact with plastic materials that have been reported to contain RMB. RMB is used frequently as an antioxidant in synthetic material. When these patients stopped smoking, the skin lesions cleared. However, consumption of sweets caused recurrences. CONCLUSION: The evaluation of reactions to single constituents of BP by testing with the special BP series facilitates understanding how sensitization may be acquired. The allergen may prove to be BP itself or 1 or more of its constituents. Testing for the constituents of this series may provide patients with a more specific allergen diagnosis and may facilitate improved therapy. BP may function as an important indicator for contact allergy to RMB. PMID- 11381346 TI - Self-reported hand dermatitis in California veterinarians. AB - BACKGROUND: Veterinarians are exposed to allergens, dirt, and chemicals, but there are very few epidemiologic data on dermatoses veterinarians. OBJECTIVE: The aim of this study is to get information about skin diseases in veterinary work. METHODS: A sample of California veterinarians were sent a questionnaire, which was returned by 73% (n = 1,416). RESULTS: History of skin atopy was reported by 11% and respiratory atopy by 63%. Dermatoses during their career were reported by 46%, and hand and/or forearm dermatitis was reported more than once and during the past year by 22% of women and 10% of men. Dermatitis with work-related exacerbating factors was reported by 28%. Almost 1 of 5 veterinarians reported animal-related skin symptoms. Other aggravators were medications (2%), gloves (4%), and other chemicals (7% of respondents). Of those with animal-related dermatitis, 65% reported only 1 animal (dog, 66%; cat, 29%; horse, 9%; and cattle, 8%), and 66% reported the symptoms appeared in minutes after the contact. In logistic analysis, the risk factors for the appearance of hand/forearm dermatitis during the past 12 months and more than once during their career, were: history of skin atopy (odds ratio, 3.5), childhood hand dermatitis (odds ratio, 3.4), history of respiratory atopy (odds ratio, 2.0), and female sex (odds ratio, 1.9). CONCLUSIONS: Veterinarians' skin symptoms were common and often caused by immediate allergy to 1 or few animal species. PMID- 11381347 TI - Contact dermatitis due to printer's ink in a milk industry employee: case report and review of the allergen paraphenylenediamine. AB - Paraphenylenediamine is a common cause of occupational dermatoses in hairdressers, metallurgy workers, and others. Hand dermatitis developed in an employee of a milk packaging facility. The employee's hands were exposed to milk cartons embossed with wet printer's ink on a daily basis for 2 years. The worker was evaluated through a history, physical examination, and patch testing with 50 standard allergens. Patch testing revealed a positive reaction to paraphenylenediamine. The hand dermatitis resolved once the patient instituted protective measures. The worker's reaction might represent a delayed-type hypersensitivity reaction to printer's ink that contained this agent. More likely, the inciting agent was some other ingredient of the printer's ink that cross-reacts with paraphenylenediamine. PMID- 11381348 TI - Allergic contact dermatitis from acrylonitrile. PMID- 11381349 TI - Commercial availability of a house dust mite patch test. AB - The house dust mite long has been studied for its potential role in atopic dermatitis. Its part in the pathogenesis of asthma and allergic rhinitis has been studied and is accepted. Testing for the dust mite allergen in this arena is performed with radioallergosorbent tests (RASTs) and prick testing. For several reasons, including the lack of a standardized testing tool, the difficulty in removing dust from the environment and the lack of association of dust and skin disease by patients, the role of the house dust mite in the pathogenesis of atopic dermatitis and skin disease has been questioned. Chemotechnique Diagnostics (Malmo, Sweden) now provides a standardized dust mite allergen in petrolatum in 2 concentrations. Dermatophagoides mix is a 50:50 mix of Dermatophagoides pteronyssinus and D. farinae at concentrations of 20% and 30%. The availability of a standardized patch test allergen for the house dust mite provides a better tool to investigate the role of this allergen as a contactant. PMID- 11381350 TI - Nickel allergy and dyshidrotic eczema: Are they related? PMID- 11381351 TI - The path to the future leads from the past. PMID- 11381352 TI - Drug-induced congestive heart failure in breast cancer survivors. AB - Breast cancer is the most common cancer among women in the United States. The administration of certain types of chemotherapy may put breast cancer survivors at risk for late effect drug-induced congestive heart failure (CHF). This case study discusses the diagnosis, management, and follow-up of drug-induced CHF in a woman with breast cancer. Discussions of risk factors and secondary and tertiary prevention of CHF caused by anthracycline chemotherapy agents are also included. Recognition of risk factors and symptoms of CHF is essential to minimize the morbidity and mortality that accompanies cardiac failure and is an important part of the comprehensive care of breast cancer patients. Because drug-induced CHF can occur years after the administration of the offending drug, primary care providers need to be aware of the risk of this disorder among breast cancer survivors under their care. PMID- 11381353 TI - Exercise self-efficacy. AB - Practitioners have the opportunity to enhance the health of their patients by improving exercise adherence through appropriate exercise prescription and monitoring. Regular physical exercise can prevent or improve many of the chronic health conditions commonly observed in clinical practice. Unfortunately, efforts to help patients adopt an exercise program are often unsuccessful. Researchers have shown that exercise self-efficacy is an important predictor of the adoption and maintenance of exercise behaviors. Self-efficacy is the belief and conviction that one can successfully perform a given activity. Patient compliance with exercise prescriptions is more likely to be successful if exercise self-efficacy is assessed and enhanced. Several questions arise for practitioners who are encouraging their patients to exercise: What are the determinants of exercise self-efficacy? How can exercise self-efficacy be assessed in the clinical setting? What can be done to enhance exercise self-efficacy? This article reviews the research literature addressing these questions and presents tools for assessing self-efficacy and prescribing appropriate exercise in the low-active adult population. PMID- 11381354 TI - Help for colicky babies. AB - The topic of colic is not new, yet parents often receive little help during a very distressing time. Colic is a diagnosis of exclusion and the diagnosis is made only after a thorough work up to rule out pathological conditions. Several studies are summarized that address the cause and management of colic. One management method, behavioral modification, is described. This intervention uses the acronym REST for Regulate, Environment cues, Structure, and Touch and for Reassurance, Empathy, Support, and Timeout. This management strategy is helpful to decrease irritability, allowing infants to become better able to regulate their environments. In most babies, colic resolves gradually or in some cases dramatically by the age of four months. PMID- 11381355 TI - Monitoring peak flow rates as a health-promoting behavior in managing and improving asthma. AB - Asthma is the most common chronic illness of childhood. Nurse practitioners, caring for patients in a primary care setting, will frequently encounter asthma. Peak flow monitoring of pulmonary function can greatly assist in diagnosis and treatment. A review of the literature on asthma management supports the many benefits of monitoring peak flow rates. Among the benefits are providing early recognition of airflow obstruction, identifying triggers, and monitoring a patient's response to therapy. Coordinated care, asthma education, and consistent practices reduce the frequency of asthma symptoms, emergency room visits, hospitalizations, and days missed from work or school. Improved communication between the nurse practitioner and the patient can facilitate more effective management of the common chronic illness. PMID- 11381356 TI - Helicobacter pylori: diagnostics and management in peptic ulcer disease: a review. AB - The management of peptic ulcer disease (PUD) has progressed markedly over the past century. Thanks to the discovery of Helicobacter pylori, our treatment focus has shifted from antacids and H2 antagonists to proton pump inhibitors and antimicrobial agents. Despite these advances, many treatment issues remain ambiguous. Some of the most controversial issues include management of the infection at a general practice level, particularly whom to treat, which regimens are most effective, and which diagnostic criteria (if any) should be used. This article will provide a brief description of available diagnostic modalities and summarize recent research findings on optimal testing techniques and treatment in primary care. PMID- 11381357 TI - Factors influencing the antibiotic-prescribing decisions of nurse practitioners. AB - Decision-making about treatment options is a complex process that is influenced by many variables. An instrument for studying this decision-making process and the factors influencing it was designed. A survey of the antibiotic-prescribing practices of nurse practitioners (NPs) was then conducted via mail and the World Wide Web. NPs who treat adult patients were presented with scenarios in which antibiotic prescription would be an appropriate choice. Participants were asked to evaluate the clinical scenarios and describe their decision-making about treatment and the rationale for treatment. Responses were analyzed to determine the factors that played a role in antibiotic decision-making and their relative importance in that process. The ranks of categories of factors by the respondents were similar across the 4 scenarios. Antibiotic-prescribing patterns and the rationale of the antibiotic-prescribing decision of the respondents for the presented scenarios are also described. PMID- 11381358 TI - Asthma education in rural communities. AB - Asthma is a common disease of airway obstruction in school-aged children. Adequate management of asthma in children leads to fewer missed school days, fewer hospitalizations, fewer emergency room visits, and an increase in quality of life. Most asthma educational programs and evaluations have focused on urban rather than rural populations. The purpose of this study was to identify parental asthma needs, develop rural asthma education materials, and evaluate the effectiveness of these educational materials in improving the knowledge and asthma care effectiveness of parents of children with asthma in a rural community. Seven parents were contacted by telephone and administered a pre- and posttest questionnaire analyzing their level of knowledge about asthma and their quality of life. Asthma educational materials were mailed to all parents in the study before administering the posttest. Results indicated that all parents needed additional education about asthma, especially regarding medications. Pre- and posttest scores showed improvements in three areas of knowledge: long-term asthma medications, controlling roaches in the home, and daily peak flow monitoring. There was a significant improvement between pre- and posttest results from the activity domain of quality of life. Eighty-five percent of the parents reported that they had either initiated changes in their home, or planned to in the future, from reading the educational materials. The parents' response to the educational materials that they received by mail was positive, indicating that they may not have received enough information about how to care for children with asthma before our study. The data suggest that distribution of asthma educational materials in rural communities can increase parental knowledge about asthma and lead to positive changes in behavior that can improve their children's health. PMID- 11381359 TI - Preceptors: a perspective of what works. AB - This article provides a perspective of clinical preceptor contributions to advanced practice nurse education. It supports quality clinical education in the face of the proliferation of academic programs, and the economics of the resulting competition. The strategies of 2 universities are used as examples of how nurse practitioner programs respond to issues such as who precepts students, how preceptors are identified, what draws preceptors to the role, and the effects of students on clinic productivity. PMID- 11381362 TI - Caspase 7 downregulation as an immunohistochemical marker of colonic carcinoma. AB - Caspases play a crucial role as apoptotic effectors; their potential implication in tumorigenesis remains to be clarified. We investigated the expression and function of caspases 7, 8, and 9 in colon cancer tissues and cell lines. Immunohistochemistry (IHC) showed downregulation of caspase 7 (22 of 26 cases) and caspase 9 (12 of 26 cases) in colonic cancer samples compared with normal mucosa on the same tissue section. Caspase 8 expression was unchanged or slightly upregulated (19 of 27 cases). The combination of IHC and Western blot analysis showed expression of the proforms of caspases 7, 8, and 9 in HT29-19A and HT29 16E colonic carcinoma cell lines. Apoptosis could be induced by staurosporine in both HT29 cell lines, with a sensitivity similar to that of the HGT cell line, but lower than that of the DAUDI cell line. Apoptosis induction in HT29 cells was concomitant with processing of caspases 3, 7, 8, and 9 and was inhibited by the caspase inhibitor ZVAD. Our data show that (1) human colon cancer cells downregulate caspase 7 and, to a smaller extent, caspase 9 in vivo and (2) in vitro staurosporine-induced apoptosis of colonic cancer cells involves caspases 7 and 9. Caspase 7 deficiency thus appears as a new immunohistochemical marker of colonic neoplasia; its correction represents a potential basis for new therapies. PMID- 11381363 TI - Progression from superficial to invasive carcinoma of the bladder: genetic evidence of either clonal heterogeneous events. AB - Until now, no definitive molecular evidence proving or disproving a true progression from superficial to invasive bladder tumors has been reported. A total of 36 lesions from 6 patients affected by invasive bladder cancer after multiple superficial recurrences were analyzed for loss of heterozygosity on 8 loci of chromosome 9 and 2 loci of chromosome 17. In addition, the clonal composition of the tumors from two female patients was examined using the human androgen receptor assay. Our data suggest that papillary bladder lesions can and sometimes do make a true progression into invasive life-threatening tumors; however, this progression is not an invariable sequence because it was definitely proven in 2 but not confirmed in 3 of the cases we examined. PMID- 11381364 TI - Loss of heterozygosity at 11q23.3 in vasculoinvasive and metastatic squamous cell carcinoma of the cervix. AB - We have previously demonstrated a strong relationship between loss of heterozygosity (LOH) at chromosome 11q23.3 and the presence of extensive tumor plugs in lymphvascular spaces (LVS) in stage 1B cervical carcinoma, suggesting that genes at this locus may regulate vasculoinvasion. This study examined LOH at 11q23.3 in microdissected tumor plugs within LVS and in metastatic foci in lymph nodes (MFLN), as well as corresponding invasive tumor and adjacent cervical intraepithelial neoplasia (CIN) 3 in stage 1B squamous cell carcinoma. Of 49 invasive carcinomas, 38.8% had LOH at 11q23.3. Of 36 tumor plugs in LVS, 39% had LOH at 11q23.3. Twenty percent of 15 MFLN demonstrated LOH at 11q23.3. Patients with LOH at 11q23.3 are significantly more likely to have disease recurrence than patients without LOH at 11q23.3 (P =.02). Of 10 foci of CIN 3, none showed LOH at 11q23.3. Although unlikely to have an impact early in carcinogenesis, tumor suppressor genes located in the region of 11q23.3 appear to be important in tumor progression, facilitating lymphvascular space invasion and, by inference, spread to lymph nodes in squamous cell carcinoma of the cervix. PMID- 11381365 TI - Histologic and immunophenotypic classification of cervical carcinomas by expression of the p53 homologue p63: a study of 250 cases. AB - Recent studies of the p53 homologue p63 indicate that this gene is preferentially expressed in basal and immature cervical squamous epithelium. This study correlated p63 expression with morphologic phenotype and human papillomavirus (HPV) type in a wide range of cervical neoplasms. Two hundred fifty cases of cervical carcinoma, including squamous cell carcinoma (SCCA; n = 178), adenocarcinoma (ADCA; n = 28), adenosquamous carcinoma (ASCA; n = 8), neuroendocrine carcinoma (NECA; n = 15), and other variant or mixed types (n = 21) were studied. Ninety-seven percent of SCCA, 0% of ADCA, and 0% of SCUC showed strong (>75% v <30%) positivity for p63 (P<.001). p63 sharply distinguished SCCA (p63+) from ADCA (p63-), Large-cell, poorly differentiated carcinomas were distinguished as putative glandular (glassy cell) or squamous (lymphoepithelial like or spindle cell) types based on p63 staining. Eight (73%) of 11 neuroendocrine tumors tested were chromogranin positive; all showed no or low (<30%) levels of p63 immunostaining. Absence of p63 was also associated with a subset of nonneuroendocrine undifferentiated carcinomas. Transitions from squamous to columnar or undifferentiated morphology coincided with loss of p63 expression. A strong association between HPV 16 and p63 positivity was identified because of the colocalization of both within tumors of squamous phenotype. p63 is a powerful marker for squamous differentiation and, when diffusely expressed, excludes a glandular or neuroendocrine differentiation. p63 may be useful for differentiating pure squamous or glandular from adenosquamous carcinomas, tracking shifts in differentiation within tumors, supporting (by its absence) the diagnosis of neuroendocrine carcinomas, and clarifying the spectrum of poorly differentiated carcinomas lacking either squamous or neuroendocrine differentiation. PMID- 11381366 TI - Apocrine ductal carcinoma in situ of the breast: histologic classification and expression of biologic markers. AB - Apocrine ductal carcinoma in situ (ADCIS) has been called a special type of ductal carcinoma in situ (DCIS) because the histologic grading is considered difficult using the classification schemes that have been proposed for common DCIS. However, ADCIS encompasses a spectrum of lesions with different morphologic aspects ranging from minimally atypical to overtly malignant. To define a classification scheme for ADCIS, 35 cases (22 pure and 13 associated with invasive carcinoma) were selected on the basis of conventional morphology on hematoxylin and eosin (H&E)-stained sections. Each case was assigned to 1 of 3 histologic grades (low, intermediate, and high) based on nuclear morphology and the presence of necrosis. In addition, the expression of hormone receptors p53, bcl-2, c-erbB-2, and Ki-67 was evaluated by immunohistochemistry, and the DNA ploidy was determined by image cytometry. Fifteen cases were classified as high histologic grade, 10 as low histologic grade, and the other 10 as intermediate grade. All but 4 cases, irrespective of grade, had the same hormonal immunophenotype: androgen receptor positivity (97.1%) and estrogen receptor and progesterone receptor negativity (94.3% and 97.1% respectively). Twenty-one cases (61.8%) showed p53 expression, and 47.1% of the cases were positive for c-erbB-2. The median positivity for Ki-67 was 5.2%. ADCIS has a unique morphologic and hormonal profile, distinct from common DCIS, deserving a specific classification. The proposed classification scheme allows for categorization of ADCIS according to the most important morphologic features already seen in common DCIS, ie, nuclear grade and necrosis. The expression of biologic markers other than hormonal receptors and bcl2 in ADCIS seems in general to be similar to that in common DCIS. Ki-67 and c-erbB-2 are expressed more frequently in intermediate and high histologic grade ADCIS. PMID- 11381367 TI - Comparative analysis of sampling methods for grossing radical prostatectomy specimens performed for nonpalpable (stage T1c) prostatic adenocarcinoma. AB - Scant data are available comparing sampling methods of radical prostatectomy specimens performed for clinical stage T1c (nonpalpable) cancer. Seventy-eight stage T1c radical prostatectomies that had 1 or more of the following adverse pathologic findings-Gleason score > or = 7, positive margins, and extraprostatic extension-were compared using 10 different sampling techniques. Of the 78 entirely submitted cases, 52 had Gleason score > or = 7, 14 had positive margins, and 54 had extraprostatic extension (mean 34 slides). Of the partial sampling methods, we favor the following two methods. The first is submitting every posterior section plus 1 midanterior section from right and left sides; if either of these anterior sections show sizeable tumor, all ipsilateral anterior slides are examined. This method detects 98% of tumors with Gleason score > or = 7, 100% of positive margins, and 96% of cases with extraprostatic extension (mean 27 slides). The second method is to use the above method but restrict it to sections ipsilateral to the previous positive needle biopsy. This method detects 92% of tumors with Gleason score > or = 7, 93% of positive margins, and 85% of cases with extraprostatic extension (mean 17 slides). Partial sampling can detect important prognostic parameters. By balancing the extra expense and time involved to process and examine additional sections with the risk of missing important prognostic parameters, pathologists can decide which sampling method to use. PMID- 11381368 TI - Isosporosis and unizoite tissue cysts in patients with acquired immunodeficiency syndrome. AB - Isospora belli, a coccidian parasite in humans, has been described as causing chronic diarrhea and acalculous cholecystitis in patients with the acquired immunodeficiency syndrome (AIDS). Diagnosis can be made at the tissue level in the epithelium of the small bowel and by fecal examination. Disseminated extraintestinal forms are uncommon. We studied 118 adult patients with AIDS and chronic diarrhea using stool analysis and endoscopy with duodenal biopsy specimen collection. These samples were processed by routine histology and transmission electron microscopy. Isosporosis was diagnosed in 8 cases. In 2 of them, unizoite tissue cysts were present in the lamina propria, with negative results in stool materials. The cysts were located within a large parasitophorous vacuole. There were no structural means of differentiating the species level of Isospora based on morphology using light or electron microscopy. We believe further work should be done to determine if unizoite tissue cysts are part of the cycle of I belli or of other species of Isospora that could be pathogenic in immunocompromised hosts. PMID- 11381369 TI - Low-grade myxoid renal epithelial neoplasms with distal nephron differentiation. AB - We report 4 distinctive renal epithelial neoplasms that are essentially identical at the morphologic and immunohistochemical levels and do not fit an accepted category in the existing classification of these lesions. The patients were all females, with ages ranging from 32 to 79 years (mean, 50 years). The tumors were well circumscribed and were composed of uniform, predominantly low cuboidal cells with eosinophilic, focally vacuolated cytoplasm. Tumor cells generally formed interconnecting tubules, with smaller areas of cordlike growth and spindling in a bubbly, myxoid stroma. All tumors were confined to the kidney, and all were immunoreactive for high-molecular-weight cytokeratin 34betaE12, cytokeratin 7, epithelial membrane antigen, and cytokeratin cocktail AE1/3. Only 1 tumor was focally immunoreactive for Ulex europaeus agglutinin. Ultrastructural study showed tumor cells forming tubular structures reminiscent of the loop of Henle or distal convoluted tubule. Follow-up in all 4 cases was benign. These distinctive tumors may be confused with aggressive sarcomatoid renal cell carcinomas because of their spindled morphology. The morphologic, immunohistochemical, and ultrastructural features of these lesions indicate differentiation toward distal nephron segments. Similar tumors probably have been reported among low-grade collecting duct carcinomas or tumors "possibly related to the loop of Henle." PMID- 11381370 TI - Mixed epithelial and stromal tumor of the kidney lacks the genetic alterations of cellular congenital mesoblastic nephroma. AB - Mixed epithelial and stromal tumor of the kidney is a recently recognized neoplasm that occurs almost exclusively in perimenopausal women. Because it frequently contains areas of smooth muscle in which epithelial structures are embedded, some have concluded that it is the adult form of congenital mesoblastic nephroma. Others have concluded that the morphology and epidemiology of mixed epithelial and stromal tumor indicate that it is unrelated to congenital mesoblastic nephroma. Although the genetic alterations of mixed epithelial and stromal tumor have not been previously elucidated, much is known about the genetic alterations of cellular congenital mesoblastic nephroma. The present study was undertaken to determine if mixed epithelial and stromal tumors have any of the genetic alterations recognized as typical of cellular congenital mesoblastic nephroma. RNA extraction was performed on formalin-fixed, paraffin embedded tissue from 7 mixed epithelial and stromal tumors followed by reverse transcription polymerase chain reaction to detect the ETV6-NTRK3 gene fusion. Fluorescent in situ hybridization with centromere-specific probes for chromosomes 8, 11, and 17 was performed to evaluate polyploidy of these chromosomes in 11 cases of mixed epithelial and stromal tumor. None of the mixed epithelial and stromal tumors showed any of these genetic alterations. We conclude that mixed epithelial and stromal tumor of the kidney lacks the genetic alterations typical of cellular congenital mesoblastic nephroma, is unrelated to it, and the appellation "adult mesoblastic nephroma" should not be used for these tumors. PMID- 11381371 TI - Increased D allele frequency of the angiotensin-converting enzyme gene in pulmonary fibrosis. AB - An insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme has previously been studied extensively in relationship to cardiovascular and renal disease. The deletion/deletion (D/D) genotype is associated with a poor outcome in immunoglobulin (Ig) A nephropathy. However, the association of this genetic marker in cardiovascular and renal disease has generated controversy, with the exception of the rate of progression and therapeutic responsiveness in IgA nephropathy. Many of the same cytokines and polypeptide mediators involved in fibrosis of the cardiovascular and renal systems have been shown to be involved in pulmonary fibrosis. We examined the I/D polymorphism of the angiotensin converting enzyme in a group of 24 patents with interstitial pneumonia and moderate to severe pulmonary fibrosis defined by radiographic studies, pulmonary function tests, and histologic findings. The incidence of the D allele in this study population was 69.0%, which is approximately 15.0% higher than the incidence in the general population of 54.0%. The incidence of the D/D genotype was 42.0%, which is approximately 11.0% greater than that in the general population (31.0%). The distribution of the D/D, I/D, and insertion/insertion genotypes of these 24 patients was not significantly different from that of historical controls (P =.1; chi(2) test); there were marginally significantly more D alleles among the 48 observed alleles than would be expected (P =.04). PMID- 11381372 TI - Calretinin, thrombomodulin, CEA, and CD15: a useful combination of immunohistochemical markers for differentiating pleural epithelial mesothelioma from peripheral pulmonary adenocarcinoma. AB - The distinction between pleural epithelial mesothelioma and peripheral lung adenocarcinoma involving the pleura is still an important diagnostic problem for surgical pathologists. The aim of our study was to identify the most specific and sensitive markers for the positive identification of mesothelioma to select a limited, appropriate panel of antibodies to differentiate between mesothelioma and adenocarcinoma. Forty-two cases of epithelial mesotheliomas and 23 cases of pulmonary adenocarcinomas were stained with the following antibodies: anticalretinin, antithrombomodulin, anti-CD44H, and monoclonal antibody HBME-1. We also studied the value of other markers in current use: cytokeratins AE1/AE3 and CAM5.2, epithelial membrane antigen (EMA), carcinoembryonic antigen (CEA), Ber-EP4, B72.3, and CD15. Of the mesotheliomas, 42 stained for calretinin, 39 (92.8%) for thrombomodulin, 42 stained for CD44H, and 41 (97.6%) stained for HBME 1. Among negative markers, 4 (9.5%) mesothelioma cases stained for CEA, 5 (11.9%) stained for Ber-EP4, 6 (14.2%) stained for B72.3, and 2 (4.7%) stained for CD15. Of the lung adenocarcinomas, 2 (8.7%) cases showed reactivity for calretinin, 5 (21.7%) for thrombomodulin, 13 (56.5%) for CD44H, all for HBME-1, 22 (95.6%) for CEA, 22 (95.6%) for Ber-EP4, 8 (34.7%) for B72.3, and all for CD15. In conclusion, calretinin and thrombomodulin were the most specific positive mesothelial markers, whereas CD44H and HBME-1 showed high sensitivity but very low specificity. Among negative markers, we advocate the use of CEA and CD15 which were the most specific in differentiating mesotheliomas from adenocarcinomas. PMID- 11381373 TI - Alterations of cell cycle regulators affecting the RB pathway in nonfamilial retinoblastoma. AB - We undertook the present study to examine alterations affecting the RB pathway in the G1 checkpoint and to determine their potential clinical significance in children affected with nonfamilial retinoblastoma. Using immunohistochemistry, patterns of expression of pRB, p16/INK4A, and E2F1 were analyzed in tissue from a cohort of 86 well-characterized patients with nonfamilial retinoblastoma diagnosed at the "Instituto Nacional de Pediatria" in Mexico City. The relationship of these phenotypes to proliferative index was assessed by analysis of Ki67 antigen expression. pRB expression was found in 11 (13%) cases. Using a hypophosphorylated specific pRB antibody, we observed low levels of underphosphorylated pRB expression in only 1 of 9 evaluable positive cases. These data suggest that the detected pRB products were hyperphosphorylated and thus had decreased functional activity. Increased p16 nuclear expression was found in only 6 tumors. No tumors showed deletions or mobility shifts of the INK4A gene. Undetectable pRB levels were significantly associated with undetectable p16 expression (odds ratio, 10.8; 95% confidence interval, 1.4-81.3; P =.03). All tumors showed nuclear immunoreactivities for E2F1 and Ki67. Increased Ki67 proliferative index was associated with increased staining for E2F1 (r =.44; P =.008) and increasing clinical stage (P =.03). Among children with unilateral disease, the mean Ki67 proliferative index was significantly higher in children with advanced clinical disease (stages 3 and 4) (mean 81.25; SD 6.78) than in those with earlier stage disease (mean 69.50; SD 9.45) (P = 0.001). Among children with bilateral disease, however, the mean proliferative index was not significantly higher for children with advanced clinical stage. When examining all cases together, there was a significant trend toward increasing proliferative index with increasing clinical stage (P =.03). In unilateral tumors, we also found that presence of detectable pRB was associated with a lower percentage of cells expressing E2F1 (46.7% v 70.8%) (P = 0.05), whereas there was no association between presence of pRB and E2F1 among bilateral tumors. We have found that expression of some of the cell cycle markers examined varies according to laterality, suggesting underlying differences in the capacity for cell cycle regulation between these 2 forms of the disease. Differences in capacities for cell cycle regulation may account for some differences in clinical behavior. Thus, the inclusion of molecular markers may become useful adjuncts to clinicopathological staging and subsequent determination of therapy. PMID- 11381375 TI - Meningeal-cutaneous relationships in anencephaly: evidence for a primary mesenchymal abnormality. AB - The pathogenesis of cranial and spinal dysraphia has been controversial. Studies of spinal dysraphia have shown that the relationships of the pia and dura to the cutaneous layers were best understood as the result of a primary abnormality of mesenchymal structures, with the nervous system lesions occurring as a result of exposure of the bare spinal cord on the body surface. This study was undertaken to determine if the relationship of the cutaneous layers in anencephaly were similar to those found in spinal dysraphia. We reviewed serial histologic sections of the cranial structures of 10 anencephalic fetuses autopsied at The Johns Hopkins Hospital. We found the dura to be continuous with the deep dermis and the pia continuous with the superficial dermis and epidermis, the same arrangement observed in myelomeningocele. The development of eyes and cranial nerves, the absence of a bony calvarium, and the meningeal-cutaneous relationships found in this study support the idea that anencephaly can originate as an abnormality of mesenchymal structures and that the brain is secondarily lost to injury in utero because of its exposed position. PMID- 11381374 TI - Immunohistochemical properties of bone marrow mast cells in systemic mastocytosis: evidence for expression of CD2, CD117/Kit, and bcl-x(L). AB - In an attempt to identify novel diagnostic markers for mast cell (MC) proliferative disorders, serial bone marrow (bm) sections of 22 patients with mastocytosis (systemic indolent mastocytosis, n = 19; mast cell leukemia [MCL], n = 1; isolated bm mastocytosis, n = 2) were analyzed by immunohistochemistry using antibodies against CD2, CD15, CD29, CD30, CD31, CD34, CD45, CD51, CD56, CD68R, CD117, HLA-DR, bcl-2, bcl-x(L), myeloperoxidase (MPO), and tryptase. Staining results revealed expression of bcl-x(L), CD68R, and tryptase in neoplastic MCs (focal dense infiltrates) in all patients. Mastocytosis infiltrates were also immunoreactive for CD45, CD117 (Kit), and HLA-DR. In most cases, the CD2 antibody produced reactivity with bm MCs in mastocytosis, whereas in control cases (reactive bm, immunocytoma, myelodysplastic syndrome), MCs were consistently CD2 negative. Expression of bcl-2 was detectable in a subset of MCs in the patient with MCL, whereas no reactivity was seen in patients with SIM or bm mastocytosis. Mastocytosis infiltrates did not react with antibodies against CD15, CD30, CD31, CD34, or MPO. In summary, our data confirm the diagnostic value of staining for tryptase, Kit, and CD68R in mastocytosis. Apart from these, CD2 may be a novel useful marker because MCs in mastocytosis frequently express this antigen, whereas MCs in other pathologic conditions are CD2 negative. PMID- 11381376 TI - Trisomy 8 as sole karyotypic aberration in an ovarian metastasizing Sertoli Leydig cell tumor. AB - Sertoli-Leydig cell tumors (SLCTs) represent a rare group of sex-cord stromal tumors of the ovary of unknown pathogenesis. We report a SLCT of intermediate differentiation with peritoneal recurrence and lymph node metastasis 12 months after removal, including cytogenetic analysis by comparative genomic hybridization and fluorescence in situ hybridization, which showed trisomy 8 as sole unbalanced karyotypic aberration. Our results provide evidence that a simple numeric chromosomal abnormality in SLCT may be associated with a malignant phenotype and suggest that the molecular pathogenesis of SLCT may be different from ovarian granulosa-stromal cell tumors. PMID- 11381377 TI - Shifting the responsibility for medical decisions. PMID- 11381378 TI - Assessment of recovery in patients undergoing intravenous conscious sedation using bispectral analysis. AB - PURPOSE: The Bispectral Index (BIS) has been recently shown to objectively predict the level of sedation in patients undergoing conscious sedation. It was the goal of this study to directly compare the recovery profile of patients where the BIS was used to monitor sedation with a control group where the monitor was not used. PATIENTS AND METHODS: Forty patients undergoing third molar extractions under intravenous conscious sedation were randomly assigned to 2 groups. In both groups, induction of sedation was performed using a standard dose of fentanyl (1.5 microg/kg) and midazolam (0.05 mg/kg). Propofol was then given in 10 to 20 mg boluses until a clinically desirable sedation level was achieved. In 1 group, the BIS was then monitored continually during surgery using a microcomputer (Aspect-1050 Monitor, Aspect Co, Natick, MA) and recorded at 5-minute intervals. The anesthetist (N.A.S.) provided additional propofol boluses to maintain a BIS level of 70 to 80. In the other group, the BIS sensor was applied, but the monitor was not used. In this group, the sedation was modified, and additional propofol was given based solely on the anesthetist's subjective assessment of the desired level of sedation (Observer's Assessment of Alertness/Sedation [OAA/S] scale level 2 to 3). Additional boluses of 1 mg of midazolam were given during the procedure if patients required repeated boluses of propofol at less than 5 minute intervals to maintain the desired sedation level (BIS level of 70 to 80 or OAA/S level of 2 to 3). These additional midazolam boluses, as well as the time of the last sedative dose (propofol or midazolam) were recorded to study the effect of these factors on recovery. RESULTS: Of the 40 patients initially included in the study, 1 subject in the BIS-monitored group was excluded due to the loss of intravenous access at initiation of the case. For the remaining 39 subjects, 19 were assessed objectively using the BIS monitor, whereas 20 were assessed subjectively using the OAA/S scale. The BIS cases were slightly longer in duration than the OAA/S cases, lasting an average of 26 minutes versus 22 minutes. This difference was statistically nonsignificant (P =.19). Less propofol was used in the BIS cases, with an average of 98 mg for BIS cases versus 106 mg for OAA/S cases (P =.59). The total dose in mg/kg/min was significantly less in the BIS group (0.054 mg/kg/min) than in the OAA/S group (0.074 mg/kg/min; P =.0082). There was no significant difference in the amount of midazolam administered after induction between the 2 groups (P =.60). The surgeon, who was blinded to whether the monitor was used, ranked the third molar extractions more difficult in the BIS group (P =.05). However, patients in the BIS group were on average more cooperative, with better maintenance of muscle tone. The difference in these parameters were nonsignificant (P =.15 and .092, respectively). A positive Romberg test was obtained earlier in BIS patients, although this difference was nonsignificant (P =.097). The straight-line test was completed significantly sooner in BIS patients (P =.013). There was no significant difference between the BIS and OAA/S groups in perceptual speed (P =.55) or computation (P =.32). There was essentially no difference between groups in patient-assessed comfort or recall of the procedure. There were also no notable differences in anesthesia complications, return to activities of daily life, or pain medication use between the 2 groups. CONCLUSIONS: The BIS provides additional information for standard monitoring techniques that helps guide the administration of sedative-hypnotic agents. It appears that use of the BIS monitor can help to titrate the level of sedation so that less drugs are used to maintain the desired level. The trend toward an earlier return of motor function in BIS-monitored patients warrants further investigation. PMID- 11381380 TI - Efficacy of temporomandibular joint arthrocentesis with and without injection of sodium hyaluronate in treatment of internal derangements. AB - PURPOSE: This study was designed to investigate the efficacy of arthrocentesis with and without injection of sodium hyaluronate (SH) into the upper joint space in the treatment of temporomandibular joint (TMJ) internal derangements. PATIENTS AND METHODS: Forty-one TMJs in 5 males and 26 females aged 14 to 53 years comprised the study material. The patients' complaints were limited mouth opening, TMJ pain and tenderness, and joint noises during function. Patients were randomly divided into 2 groups in which only arthrocentesis was performed in 1 group and arthrocentesis plus intra-articular injection of sodium hyaluronate was performed in the other group. Both groups contained patients with disc displacement with reduction and with closed lock. Clinical evaluation of the patients was done before the procedure, immediately after the procedure, on postoperative day 1, and at 1, 2, 3, 4, 5, 6, 9, 12, 18, and 24 months postoperatively. Intensity of TMJ pain, jaw function, and clicking sounds in the TMJ were assessed using visual analog scales. Maximal mouth opening and lateral jaw movements also were recorded at each follow-up visit. RESULTS: Both techniques increased maximal mouth opening, lateral movements, and function, while reducing TMJ pain and noise. CONCLUSIONS: Although patients benefitted from both techniques, arthrocentesis with injection of SH seemed to be superior to arthrocentesis alone. PMID- 11381382 TI - The relationship between surgical changes in dentofacial morphology and changes in maximum bite force. AB - PURPOSE: This study determined whether patients with greater surgical changes, and presumably greater normalization of their skeletal morphology, showed greater increases in their maximum voluntary bite forces after orthognathic surgery. PATIENTS AND METHODS: A total of 104 adult patients (32 males and 72 females) treated with 1 of 8 different orthognathic surgical procedures were examined. Patients' presurgical and postsurgical morphologic and biomechanical measurements were taken from lateral cephalograms. Measurements known to be related to maximum bite force were used in the analysis. Patients' presurgical and postsurgical maximum bite forces were measured at 8 tooth positions (ie, right and left incisors, canines, premolars and molars). RESULTS: Presurgical and postsurgical morphology and biomechanics variables were strongly correlated with each other, suggesting that orthognathic surgery produced relatively little change in patients' overall craniofacial form. Maximum voluntary bite forces were primarily correlated with variables relating to jaw size-both before and after surgery. No correlations were noted between the increases in maximum voluntary bite forces and surgically produced changes in skeletal morphology and the biomechanics variables. CONCLUSIONS: Factors other than surgically produced changes in skeletal morphology are responsible for increases in maximum voluntary bite force after orthognathic surgery. PMID- 11381383 TI - Depression, pain, exposure to stressful life events, and long-term outcomes in temporomandibular disorder patients. AB - PURPOSE: This study investigated the role of psychological factors in temporomandibular disorders (TMD). Orofacial pain patients' pretreatment levels of depression, disability caused by pain, and exposure to stressful life events were measured, and differences on these variables between temporomandibular joint (TMJ) disease patients and patients whose pain was of muscular origin (MPD) were evaluated. The use of these variables and patient diagnostic status in predicting response to treatment in a subsample of these patients was also evaluated. PATIENTS AND METHODS: Before undergoing treatment, 258 patients were administered the Beck Depression Inventory (BDI), the Pain Disability Index (PDI), and the Social Readjustment Rating Scale (SRRS). Follow-up data on pain disability, current level of pain, depression, and satisfaction with treatment were obtained on 48 of these patients who were contacted at varying intervals after completing treatment. RESULTS: BDI scores obtained at the outset of treatment were significantly elevated and were positively correlated with SRRS and PDI scores. MPD patients had higher SRRS, BDI depression, and PDI pain disability scores than TMJ patients, and differences between the 2 groups in pain disability were greatest in areas that are often sources of interpersonal stress. Among follow-up patients, PDI scores declined after treatment, with MPD patients showing greater decreases than TMJ patients. Independent of patients' diagnostic status, their pretreatment PDI scores were predictive of their pain level at follow-up and were inversely related to their degree of satisfaction with treatment at follow-up; their pretreatment BDI scores were predictive of their depression level at follow up. CONCLUSIONS: The findings are consistent with previous research indicating a link between emotional dysfunction and TMD and are largely supportive of the conclusion that psychological factors play a more pronounced role when pain is of muscular origin. Promising behavioral interventions are available for TMD patients in whom psychological factors appear to be playing a significant role. PMID- 11381385 TI - A comparison of 3 methods of face-bow transfer recording: implications for orthognathic surgery. AB - PURPOSE: The purpose of this study was to compare the occlusal plane inclination of models mounted using 3 different systems for face-bow transfer with the actual occlusal plane inclination as measured on a cephalometric radiograph. PATIENTS AND METHODS: Twenty-two subjects were enrolled in this study. Three alginate impressions of the maxillary dentition were taken, and 3 stone dental models were produced for each subject. Face-bow recordings were obtained on each subject using the SAM Anatomical Face-bow (Great Lakes Orthodontics Products, Ltd, Tonawanda, NY), the Erickson Surgical Face-bow (Great Lakes Orthodontics Products, Ltd) and a new technique developed by one of the authors (J.G.). For each subject, the dental models were mounted on a SAM articulator using each of the 3 face-bow recordings. Finally, a lateral cephalometric radiograph was obtained for each subject. The occlusal plane inclination was measured on the models and on the cephalometric radiographs. Differences among groups were tested using a 1-way analysis of variance. Bonferroni test was used for post hoc comparison between different pairs of groups. RESULTS: The average occlusal plane inclination using the SAM Anatomical Face-bow was 7.8 degrees +/- 4.2 degrees greater than the actual-a difference that was statistically significant. The mean occlusal plane inclination of the models obtained using the Erickson Surgical Face-bow was 4.4 degrees +/- 2.2 degrees greater than the actual-a difference that was also statistically significant. The mean occlusal plane inclination of the models obtained by the new technique was only 0.9 degrees +/- 1.2 degrees greater than the actual; this difference was not statistically significant. CONCLUSIONS: The new mounting technique is more accurate than the conventional SAM Face-bow or the Erickson Face-bow for reproducing the actual occlusal plane inclination. PMID- 11381387 TI - Blood supply to the platysma muscle flap: an anatomic study with clinical correlation. AB - PURPOSE: This study evaluated the blood supply of the superiorly based and posteriorly based platysma muscle myocutaneous flap. MATERIALS AND METHODS: Four fresh cadaver specimens were injected with Microfil (Flow Technologies, Inc, Carver, ME) red stain in the common carotid artery and blue stain in the brachiocephalic vein. To clarify the blood supply to the platysma muscle, the blood vessels were first identified on lateral and posteroanterior radiographs of the specimens. Subsequently, anatomic dissection of the arteries and veins associated with the platysma muscle and overlying skin was performed. RESULTS: The submental artery was the primary vessel to the platysma muscle. The superior thyroid artery, occipital artery, and posterior auricular artery were identified as secondary vessels. The external jugular vein provided the primary venous drainage, followed by the submental vein. CONCLUSIONS: The superiorly based design has a reliable arterial blood supply (facial and submental arteries) and poor venous drainage. The posteriorly based design has a reliable venous drainage (external jugular vein), but the arterial blood supply is almost random (occipital artery). PMID- 11381388 TI - The use of computed tomography to define zygomatic complex position. AB - PURPOSE: The purpose of this study was to analyze the use of midline references and landmarks to assess the position of the zygomatic complex relative to the cranial base, and to test the reliability of these measurements in assessing facial symmetry. METHODS: Direct skull measurements were compared with measurements made on computed tomography (CT) images. The effect of CT scanner error, technologist error, gantry angle error, error of skull inclination, and error due to the presence of titanium rigid fixation hardware were assessed. To test observer variation and the effect of each level of error, 4 blinded bilateral measurements were repeated 3 times by 5 observers on both dry skull and CT scans. A mixed effect analysis of variance model then assessed for effect of method of measurement (dry skull vs CT), observer, CT scanner, technologist, gantry angle, skull inclination, and rigid fixation. RESULTS: A total of 2,040 measurements were made. Measurements for zygomatic complex posterior and anterior width and height were reliable and had an interobserver variations of 0.02 +/- 0.03 mm, 0.5 +/- 0.4 mm, and 0.37 +/- 0.3 mm, respectively. The difference between dry skull and CT assessment for the 3 reliable measurements was 1.2 +/- 0.3 mm, 0.44 +/- 0.4 mm, and 1.1 +/- 0.5 mm, respectively. The errors produced by the CT scanner, technologist, and rigid internal fixation hardware were not clinically significant. The measurements were not sensitive to gantry angle and skull inclination changes of 10 degrees or less. A fourth measurement assessing zygomatic complex projection was found not to be accurate or reliable. CONCLUSIONS: These findings suggest that the 3 CT scan measurements describing the position of the zygomatic complex relative to the cranial base are clinically useful. PMID- 11381390 TI - Slowly expanding palatal mass. PMID- 11381392 TI - Giant ancient neurilemmoma of the cervical region: report of case. PMID- 11381391 TI - From the chromosome to DNA: Restriction fragment length polymorphism analysis and its clinical application. AB - Understanding how chromosomal alterations contribute to acquired and inherited human disease requires the ability to manage the enormous physical and informational complexity of the deoxyribonucleic acid (DNA) packaged within. Important concepts and techniques involved in the analysis of DNA include restriction enzymes, Southern blotting, and restriction fragment length polymorphism/linkage analysis. These techniques have been essential in the understanding and diagnosis of several syndromes associated with the head and neck. The purpose of this article is to introduce DNA structure, describe some techniques fundamental to DNA analysis, and provide a brief overview of the clinical applications of this technology with respect to dentinogenesis imperfecta and oral field cancerization. PMID- 11381393 TI - T-cell lymphoma in the parotid region after cardiac transplant: case report. PMID- 11381394 TI - Keratotic basal cell carcinoma of the upper gingiva with cervical lymph node metastasis: report of a case. PMID- 11381395 TI - Small cell carcinoma of the oral cavity: report of a case. PMID- 11381396 TI - Progressive facial hemiatrophy associated with an astrocytoma in the cerebellopontine region. PMID- 11381397 TI - Closure of a large alveolar cleft by bony transport of a posterior segment using orthodontic archwires attached to bone: report of a case. PMID- 11381398 TI - Tonsillolith: a case report. PMID- 11381399 TI - A fatal outcome from rhinocerebral mucormycosis after dental extractions: a case report. PMID- 11381400 TI - Solitary osteoma of the mandibular ramus: report of a case. PMID- 11381401 TI - Maxillary amelanotic melanoma: a case report. PMID- 11381402 TI - Use of acrylic spheres as spacers in staged temporomandibular joint surgery. PMID- 11381403 TI - Continuous postoperative pain control after bone graft harvest using a pump delivery system. PMID- 11381404 TI - A single standard in double degree. PMID- 11381405 TI - Seventh nerve injury during temporomandibular joint surgery. PMID- 11381407 TI - In memoriam: H. William Clatworthy, Jr, MD 1917-2000. PMID- 11381408 TI - Fetal tracheal occlusion in the rat model of nitrofen-induced congenital diaphragmatic hernia: tracheal occlusion reverses the arterial structural abnormality. AB - BACKGROUND/PURPOSE: The high mortality rate of congenital diaphragmatic hernia (CDH) is ascribed generally to pulmonary hypoplasia and persistent pulmonary hypertension characterized by associated pulmonary arterial structural changes. Prenatal tracheal occlusion (TO) accelerates lung growth, but the effect of TO on pulmonary arterial structure in CDH has not been well defined. The authors hypothesized that TO could reverse the pulmonary arterial structural changes observed in CDH. To address this hypothesis, we utilized the nitrofen-induced rat model of CDH to examine the effect of TO on pulmonary arterial morphology of CDH lungs. METHODS: Left-sided CDH was induced by administering 100 mg of nitrofen to pregnant Sprague-Dawley rats on day 9 of gestation. TO was performed on day 19, and the fetuses were harvested on day 21.5 of gestation. After the ductus arteriosus was ligated, the pulmonary arteries were injected with a barium gelatin mixture, and the lungs were inflation fixed. Coronal sections of the lungs were stained with elastin van Gieson. External diameter (ED), internal diameter (ID), and medial and adventitial wall thickness of the pulmonary arteries were measured using a computer image analyzer, and the percent medial thickness (%MT) and adventitial thickness (%AT) were calculated. The lungs from nitrofen-exposed fetuses with left-sided CDH (CDH group), trachea-occluded left sided CDH (CDH+TO group), non-CDH (non-CDH group), and normal fetuses (normal group) were compared. RESULTS: The %MT was significantly increased in all sizes of arteries in CDH compared with non-CDH and normal groups (P < .01). Compared with the CDH group, the CDH+TO group had significantly reduced %MT in all sizes of arteries (P < .01), to values comparable or less than the non-CDH and normal groups. The %AT of the CDH group was significantly increased in larger arteries compared with non-CDH and normal control groups (P < .01). CDH+TO had significantly decreased %AT compared with CDH in both larger (P < .01), and smaller arteries (P < .05) and that was comparable with the non-CDH and normal control groups. CONCLUSIONS: TO in hypoplastic CDH lung can reverse the pulmonary arterial structural changes that are seen in the nitrofen-induced fetal rat model of CDH. These data suggest that TO may reduce pulmonary vascular reactivity, and the risk of postnatal persistent pulmonary hypertension observed in human neonates with severe CDH. J Pediatr Surg 36:839-845. PMID- 11381409 TI - Left-lung-collapse bronchial deformation in giant omphalocele. AB - Five infants with giant omphalocele had persistent collapse of the left lung and required prolonged respiratory support. Narrowing of the left main bronchus, reversible with positive end-expiratory pressure, was identified radiographically in 3 infants, and we postulate that this relates to distortion of the bronchus within the constraints of the elongated, narrow thoracic cavity characteristic of these patients. The lung collapse may be precipitated by manipulation (reduction or attempted reduction) of the omphalocele. J Pediatr Surg 36:846-850. PMID- 11381410 TI - Usefulness of cord-blood harvesting for autologous transfusion in surgical newborns with antenatal diagnosis of congenital anomalies. AB - BACKGROUND/PURPOSE: The risks of homologous transfusion and the effectiveness of predeposit autologous transfusion have been described. The authors examined the clinical usefulness of cord-blood harvesting for autologous transfusion in newborns who had congenital anomalies antenatally diagnosed that would require surgical intervention at or near the time of delivery. METHODS: Of 112 cases of antenatal diagnosis of congenital anomalies, 50 mothers gave informed consent and enrolled in this study. Cord-blood was withdrawn immediately after clamping of the umbilical cord and was used for autologous transfusion in newborns within the first 3 days postpartum. RESULTS: A mean of 72 +/- 54 mL of cord-blood was harvested (27 +/- 18 mL/kg). While preserving cord-blood for 3 days at 4 degrees C, no signs of clot formation or hemolysis were observed. The harvested cord blood included plasma-free Hb ranging from 1 to 68 (13 +/- 18) mg/dL and thrombin antithrombin III complex ranging from 2 to 273 (18 +/- 50) ng/mL. Bacteriologic examination of the stored cord-blood showed negative cultures, except for samples from 3 newborns after vaginal delivery. A mean of 46 +/- 34 mL of cord-blood was used in 26 patients for autologous transfusion. No significant complications related to cord-blood transfusion were recognized clinically. CONCLUSIONS: Autologous cord-blood transfusion has the potential to be a useful alternative to homologous transfusion in newborns requiring surgery. Adequate collection and storage techniques for cord-blood must be developed. J Pediatr Surg 36:851-854. PMID- 11381411 TI - Circular myotomy of the distal esophageal stump for long gap esophageal atresia. AB - PURPOSE: The aim of this work is to report the utility of the distal esophageal stump's circular myotomy in case of a long gap esophageal atresia repair. METHODS: Between 1972 and 1999 the authors treated 192 patients who had esophageal atresia (EA). Among them, 6 patients with EA long gap underwent both a distal and proximal stump circular myotomy. Five cases were Gross type C, and 1 case was Gross type A. The gap (average 4.5 cm) did not permit a simple and direct end-to-end anastomosis. RESULTS: One patient died 6 days after the operation because of a cardiac malformation. There was no mucosal tear during the myotomies or any anastomotic stricture later. Five patients survived. Three of them needed an antireflux procedure (60% of surviving patients). None of the 5 patients showed any mucosal outpouching, and their esophageal motility and swallowing were not different clinically compared with the patients who underwent an EA repair without a myotomy. CONCLUSIONS: Distal circular myotomy is a very useful, however, delicate, procedure that can help solve the problem given by long gap EA. It is mandatory not to tear the mucosa during the myotomy to avoid the shortening of the stump caused by its repair, which would lead to an increase in the size of the gap. J Pediatr Surg 36:855-857. PMID- 11381412 TI - Educational attainments in early adolescence of infants who required major neonatal surgery. AB - PURPOSE: The aim of this study was to assess long-term educational attainment in adolescence of former infants who required major neonatal surgery. METHODS: The study is an extension of a prospective longitudinal study begun in 1983 on 30 full-term newborns requiring major neonatal surgery, a matched healthy control group, and full-term infants requiring neonatal intensive care for medical reasons. Educational attainment levels were obtained from the results of compulsory national curriculum examinations taken at age 11 years in 3 core academic subjects, English, Mathematics, and Science and teachers' assessments of current academic performance. Additionally, follow-up data on health were obtained from family general practitioners and parents. RESULTS: Seventy percent of the original surgical group, 48% of the original control group, and 77% of the original medical group underwent follow-up. The participants were aged between 11 and 13 years. After adjustment for social factors (gender, social group, and mother's age), the surgical group was significantly behind on all measures of educational attainment (English, P < .0005; Mathematics, P < .02; Science, P < .0005; academic performance, P = .004), compared with the control group and medical group who did not differ from each other. Independent predictors of outcome were mechanical ventilation for >or=4 days in the neonatal period and behavior problems at 3 years. At 12 months of age, independent predictors of cognitive functioning were length of hospitalization and, at 3 years of age, the number of operative procedures. Mechanical ventilation was not significantly associated with cognitive functioning at previous stages of the study. CONCLUSIONS: These results show that, in early adolescence, children who required major neonatal surgery were performing less well academically compared with their peers. This information can be used to increase awareness of the difficulties these children may experience during childhood so that intervention can be directed appropriately, thereby lessening the risk of later educational problems. J Pediatr Surg 36:858-862. PMID- 11381413 TI - Cremasteric reflex and retraction of a testis. AB - PURPOSE: A clinical study has been planned to define the clinical characteristics of cremasteric reflex (CR) for deciding on the possibility of a prolonged suprascrotal stay of a testis through this reflex. METHODS: Five hundred boys from 3 days to 16 years of age were divided into 6 groups according to their ages and were evaluated for the presence of the CR. After evoking CR, the presence or absence of changes in testicular location and the most elevated position of the testis were recorded. Testicular position difference (TPD), the duration of the stay of testis in the most elevated position (DEP), and the interval for reevoking the CR (IRCR) were determined. The number of consecutive responses after repetitive evokings were recorded as reproducibility (R). Mean TPD, DEP, IRCR, and R values for both sides were calculated and compared among groups. The boys with a positive reflex were classified further according to their TPD, DEP, IRCR, and R values. Three groups were established according to the aforementioned criteria by dividing the values into 3 equal parts. The association of each of the parameters to other parameters were compared. RESULTS: Bilateral positive CR was encountered in 42.7% of newborns, 36.3% of the boys between 1 month and 1 year old, 38.1% of the boys between 2 years and 4 years old, 75.2% of the boys between 5 years and 8 years old, 70.3% of the boys between 9 years and 12 years old, and 72.1% of the boys between 13 and 16 years old. The highest percentage of the contralateral activations during ipsilateral evokings were encountered in boys who were between 5 and 8 years of age. The highest mean TPD and mean R, the longest mean DEP, and mean IRCR were encountered in boys between 5 and 8 years of age. Boys with the highest TPD did not have shortest IRCR and highest R values. Similarly, boys with the longest DEP or shortest IRCR and highest R values did not have the association of other parameters that would suggest a hypersensitivity. CONCLUSIONS: The rate of presence and the characteristics of a positive CR vary largely. However, suprascrotal location of a testis for extended periods through the activation of this reflex does not seem to be likely. Instead of a hyperactive reflex, the clinical condition, so called the retractile testis, might have resulted through alterations within the cremaster muscle itself. J Pediatr Surg 36:863-867. PMID- 11381414 TI - Minimally invasive laparotomy for treatment of neonatal ovarian cysts. AB - BACKGROUND/PURPOSE: The optimal management of neonatal simple ovarian cysts greater than 4 cm is not known. The authors describe a safe, simple, definitive surgical technique. METHODS: We report 5 cases of simple ovarian cysts greater than 4 cm treated at the authors' institution between 1998 and 1999. All initial ultrasound findings were consistent with simple ovarian cysts greater than 4 cm. All patients underwent minimally invasive laparotomy. Length of operation ranged between 28 and 49 minutes. There were no intraoperative nor postoperative complications. The operation did not delay discharge. All patients were taking enteral nutrition and required no analgesia by postoperative day 1. No patient had recurrence of the ovarian cysts when screened by ultrasound scan at 4 months of age. CONCLUSION: Minimally invasive laparotomy provides a safe, definitive treatment for neonatal ovarian cysts greater than 4 cm. J Pediatr Surg 36:868 869. PMID- 11381415 TI - Surgical treatment of the Mullerian duct remnants. AB - BACKGROUND: Persistent Mullerian duct tissue in male individuals may result in an enlarged prostatic utricle (utricular cysts and utricle) or a Mullerian duct cysts, either distinctively or synonymously. In intersex patients Mullerian duct remnants (MDR) are an usual occurrence. Surgical excision is the definitive treatment of symptomatic remnants, as well as during the reconstruction of intersexual genitalia. Many approaches have been described. The authors review their experience in intersex patients. METHODS: From 1986 to 1999, the authors treated 111 patients with intersex disorders. The records of 47 patients raised as boys with MDR were reviewed. Based on the symptoms and the size of the remnants, in 32 patients the structures were removed. In 13 patients extirpation was done by perineal approach, in 9 by transperitoneal approach, and in the remaining 9 patients the combined abdominal and perineal approach were undertaken. In one patient the large prostatic utricle was extirpated by a posterior sagittal pararectal approach. Perineal approach was mainly used in younger asymptomatic children, with the prostatic utricle disclosed incidentally during genitography because of intersex disorders. Operation was performed only in cases in which the prostatic utricle was observed by cystoscopy or identified by Fogarty balloon catheter introduction into the prostatic utricle. In older patients these structures were discovered frequently after failed urethroplasty, or after symptoms of urinary infection, urinary retention, or epididymitis. We elected to use the transperitoneal approach based on the extension of these structures into the pelvis. The average age of patients at the time of surgery was 8.6 years, with a range of 1 to 30 years. RESULTS: There were no rectal or bladder injuries during surgery. An older patient had temporary impotence after abdomino-perineal extirpation. The lack of ejaculation, seen in 5 patients, was related to frequent intra-utricular termination of the vas deferens. Posterior sagittal pararectal approach certainly enabled complete exposure and exact visualization of all structures, with considerably decreased bleeding. If gonadal biopsy or gonadectomy were necessary, the transperitoneal approach could not be avoided. CONCLUSIONS: Surgical treatment of MDR in intersex patients varies according to the size of the utricle, and a double approach is often necessary. A high degree of success may be achieved with minimal morbidity. J Pediatr Surg 36:870-876. PMID- 11381416 TI - Cremaster muscle is not sexually dimorphic, but that from boys with undescended testis reflects alterations related to autonomic innervation. AB - BACKGROUND/PURPOSE: The genitofemoral nerve and its motor neuron are known to be sexually dimorphic. An association has been suggested to exist between antiandrogen-induced undescended testis and the genitofemoral nerve. Therefore, the cremaster muscles (CM) from boys and girls with inguinal hernia and boys with undescended testis were compared to evaluate the failed descent through sexual dimorphism. METHODS: Samples of CM were obtained from 10 boys and 10 girls with inguinal hernia and 10 boys with undescended testis. Sections were stained for ATPase reactions after acid and alkaline preincubations, and for the expression of myosine heavy chains. The percentages of type 1 and type 2 fibers, and mean diameters of fibers according to the types were determined for each group. Values were compared through Mann-Whitney U test, and P values less than .05 were considered to be significant. RESULTS: CM have mainly been composed of type 1 fibers. Distributions of type 1 and 2 fibers have not shown a difference among groups. Mean diameters of both type 1 and type 2 fibers from girls (P < .05), and mean diameter of type 2 fibers from boys with undescended testis have been significantly smaller than those encountered in boys with inguinal hernia (P < .05). CONCLUSIONS: Although the mean diameter of type 1 fibers from boys with undescended testis has been as large as those encountered in boys with inguinal hernia, mean diameter of type 2 fibers has been as small as those encountered in girls. Among various reasons to selectively diminish the size of type 2 fibers, lack of beta adrenergic effect appears to be the most satisfactory explanation. J Pediatr Surg 36:877-880. PMID- 11381417 TI - Routine intraoperative cholangiography during laparoscopic cholecystectomy minimizes unnecessary endoscopic retrograde cholangiopancreatography in children. AB - PURPOSE: The aim of this study was to determine the necessity for intraoperative cholangiography (IOC) during pediatric laparoscopic cholecystectomy (LC). METHODS: A retrospective review of 100 consecutive patients undergoing LC was conducted. RESULTS: Ninety-eight children underwent successful LC. The average age was 11.3 years. IOC was successful in 55 of 63 studies. Operating time for patients with IOC averaged 91 minutes, and without IOC, 67 minutes. Twenty children had preoperative ultrasound, laboratory, or clinical evidence of common bile duct (CBD) stones. Fifteen of these 20 children actually had CBD stones. Three additional children who lacked any ultrasound, clinical, or laboratory evidence of choledocholithiasis had unsuspected CBD stones. Eight children, therefore, had ultrasound, clinical, or laboratory findings not predictive of the actual state of the CBD. Sixteen children underwent endoscopic retrograde cholangiopancreatography (ERCP), 9 preoperatively and 7 postoperatively. Four preoperative ERCP studies showed no CBD stones. There were no complications from performing IOC. CONCLUSIONS: (1) CBD stones are common in children with gallstones, (18 of 100 patients). (2) Preoperative studies and clinical findings may not predict accurately the presence or absence of CBD stones. (3) IOC should be routinely performed in children before the use of ERCP to avoid unnecessary ERCP unless CBD stones are specifically visualized by ultrasound scan. J Pediatr Surg 36:881-884. PMID- 11381418 TI - Mutational analysis of the BMP-1 gene in patients with gastroschisis. AB - BACKGROUND: Gastroschisis is a rare abdominal wall defect. Although the pathogenesis of gastroschisis is unknown, there is some evidence of the genetic etiology of gastroschisis. Recently, a functionally null deletion of the mouse bone morphogenic protein-1 (BMP-1) gene resulted in a phenotype that resembled a human neonate with gastroschisis. BMP-1 thus became the first potential candidate gene for gastroschisis. METHODS: To explore this possibility the authors collected blood samples from 11 patients who had gastroschisis. Mutational analysis of exons 2 to 15 of the human BMP-1 gene was performed using genomic polymerase chain reaction, single-strand conformation polymorphism analysis and direct sequencing methods. RESULTS: No mutation of the human BMP-1 gene was observed in any of these patients. CONCLUSION: Although heterogeneous etiologies might be proposed for gastroschisis, our results provide further evidence of a nongenetic etiology for gastroschisis. J Pediatr Surg 36:885-887. PMID- 11381419 TI - Survival and outcome after hepatic artery thrombosis complicating pediatric liver transplantation. AB - BACKGROUND/PURPOSE: Hepatic artery thrombosis (HAT) represents a significant cause of graft loss and mortality after pediatric orthotopic liver transplantation (OLT). The incidence and etiology of this complication have been investigated in detail but relatively little is known about outcome. METHODS: A review was conducted of all children with confirmed HAT complicating OLT during a 10-year period (1990 through 1999) in a single center. HAT was established by angiography or at operation in all cases. RESULTS: From a consecutive series of 400 pediatric OLTs, there were 31 (7.8%) instances of HAT in 29 children of median age 3.8 years (range, 8 days to 16 years). Twenty-four (83%) are alive after a median follow-up of 3.6 years. Fourteen cases occurred after transplantation of whole grafts and 17 after reduced or split livers. Of the 18 episodes resulting in retransplantation, there were 5 deaths and 2 second episodes of HAT; surviving children are alive with good graft function. Of the 13 episodes managed without retransplantation, 4 patients underwent attempted early revascularisation of the graft, which was successful in 2, and the remainder initially were treated conservatively. All 13 children are alive after a median follow-up of 4.1 years (range, 0.6 to 5.8), but 5 required radiologic or surgical intervention for biliary or septic complications; biochemical liver function is normal in 8, mildly abnormal in 3, and poor in 2. Retransplantation was less likely in those who had received reduced or split grafts (7 of 17) compared with those who had received whole grafts (11 of 14), but this difference just failed to reach statistical significance (chi(2) = 3.01, 0.1 > P > .05). CONCLUSIONS: Using a selective policy of retransplantation, revascularisation, and conservative treatment, 83% of children survived HAT complicating OLT. Approximately 40% of children with HAT survived without retransplantation. J Pediatr Surg 36:888-891. PMID- 11381420 TI - Survival patterns in biliary atresia and comparison of quality of life of long term survivors in Japan and England. AB - BACKGROUND/PURPOSE: Portoenterostomy is an accepted method of achieving bile drainage in biliary atresia, but there is a paucity of data, including formal quality-of-life (QoL) studies, on long-term survivors. This report includes survival analysis and QoL studies from the world's largest series of cases treated in Japan (1951 to 1998). The Japanese QoL results are compared with a matched group of UK patients from King's College Hospital, London. METHODS: One hundred fifteen Japanese surviving portoenterostomy patients were studied and comparison of trends in survival calculated from 6-year period cohorts. Liver function and hematologic status in a group of 30 long-term survivors (14 to 24 years) were compared with 25 patients from England, (14 to 23 years). Twenty-five Japanese and 21 UK patients (SF-36) completed a QoL questionnaire. RESULTS: Median survival times in Japanese patients before 1975 were less than 1 year but increased to 18 years after 1975. Hematologic and liver function test results did not show any significant differences between the Japanese and UK patients. QoL studies in the UK patients showed no significant difference from normative, general population data. Japanese patients underperformed in general health (P = .01), role emotional (P = .05) and role physical (P = .07) but, overall, there was no significant difference between the Japanese and UK patients except for marginal differences in indices of general health and vitality (P = .06 and .04, respectively). CONCLUSIONS: Long-term survival rate in the Japanese patients increased dramatically from 1 year to 17 years after 1975. The QoL of survivors was comparable in Japan and England. The satisfactory comparison with normative population data suggests that we should continue to use portoenterostomy as the primary treatment for biliary atresia. J Pediatr Surg 36:892-897. PMID- 11381421 TI - Clinicopathologic relationship of hypoganglionosis. AB - BACKGROUND/PURPOSE: Congenital motor dysfunction of the intestine associated with a morphologically abnormal myenteric nervous plexus (MP) is known as Hirschsprung's disease allied disorder (HAD). However, the clinicopathologic features of HAD are not well understood, partially because a standardized method of histologic evaluation of MP has not been established. To elucidate the clinicopathologic relationship of HAD the authors reviewed 6 cases of HAD using a newly devised histologic evaluation method. METHODS: Flat-mounted frozen sections of the ileum were stained for S-100 protein by fluorescent immunohistochemistry. Quantitative evaluation of MP was performed by measuring the fluorescence positive area (MP ratio), and the results were compared with those of age-matched normal controls. RESULTS: All of 6 patients required laparotomy within 1 month after birth and enterostomy between 23 days and 10 months. Three died of intractable enteritis by the age of 2.2 years and were totally dependent on parenteral nutrition (PN) throughout their lives. The other 3 have survived for 6 to 10 years but have required PN occasionally. MP ratio in controls was more than 0.34 at all ages, whereas that in HAD was significantly lower than that in controls according to the clinical severity. CONCLUSION: MP size measured on 2 dimensional demonstration is suggested to be an indicator of clinical severity of HAD. J Pediatr Surg 36:898-900. PMID- 11381422 TI - Renal pelvic pressure responds with augmented increases to increments in intraabdominal pressure. AB - BACKGROUND/PURPOSE: Flow of a fluid through a collapsible tube is under the influence of various factors including the external compressing pressure. Because the intraabdominal pressure may compress the ureter, an experimental study has been planned to determine and compare the normal intraabdominal and renal pelvic pressures and the alterations in renal pelvic pressure in response to the increments in intraabdominal pressure in the rabbits. METHODS: Eight adult rabbits were used for the experiment. Under general anesthesia, an urethral catheter, a nasogastric tube, and an intraperitoneal catheter were placed to measure intravesical (IVP), intragastric (IGP), and intraabdominal pressures (IAP), respectively. Intracranial pressure monitorization catheter was placed into the renal pelvis to monitor intrapelvic pressure (IPP). Basal pressure measurements have been recorded. The pressures have been recorded in every 5 minutes, and IAP has been increased gradually about 3 to 4 cm of water pressure in every step for 30-minute periods. RESULTS: Increases in the intrapelvic pressure values have been significantly higher than the increases in the IAP (P < .001). A significant correlation has been found between IPP and IAP (P = .000, r = 0.866). By using linear regression analysis the relationship has been found to be IPP = 7.303 + 1.985 (IAP). Intragastric pressure values have been higher compared with IAP values (P < .001), whereas intravesical pressures have not differed from IAP (P > .05). CONCLUSIONS: Elevations in IAP results in augmented increases in the IPP. Poiseuille and Laplace Laws suggest this augmented increase to resemble proximal ureteric obstruction. Increases in IAP may simulate proximal ureteric obstruction and may take part in the pathogenesis of hydronephrosis. J Pediatr Surg 36:901-904. PMID- 11381423 TI - Lipoblastoma. AB - BACKGROUND/PURPOSE: Lipoblastoma is a rare benign mesenchymal tumor of embryonal fat that occurs almost exclusively in infants and children. This study was done to determine the clinical and pathologic characteristics of lipoblastoma. METHODS: Nine cases of pathologically proven lipoblastoma from 1979 to 1997 were reviewed. There were 6 boys and 3 girls ranging in age from 3 months to 29 months. RESULTS: A soft tissue mass was the chief complaint in 7 patients, abdominal distension in 1 patient with a retroperitoneal mass, and defecation difficulty in 1 patient with a perirectal mass. In 4 patients, tumors occurred on the back. Other tumor location includes the neck, scrotum, retroperitoneum, perirectal area, and buttock in 1 patient each. Lesions measured 2.3 to 19.5 cm. Complete excision was done in 8 patients. One perirectal tumor was removed by both the posterior sagittal approach and the intraabdominal approach but incompletely resected. Two tumors located on the back recurred with intraspinal extension 12 months and 18 months after resection. Second resection and second resection with laminectomy were done. Leg pain and urinary incontinence developed in 1 patient but improved on conservative treatment. CONCLUSIONS: Lipoblastoma is a benign neoplasm but can frequently recur (25%) in spite of complete excision. Lipoblastomas occurring on the back had a high recurrence rate (50%) and associated with intraspinal extension. J Pediatr Surg 36:905-907. PMID- 11381424 TI - Inflammatory myofibroblastic tumor in children: diagnosis and treatment. AB - BACKGROUND/PURPOSE: Inflammatory myofibroblastic tumor (IMT) is a rare benign neoplasm. Although it is commonly seen in children, the number of childhood cases in the current literature is limited. Furthermore, malignant degeneration or transformation to lymphoma in the recurrent or residual IMT have directed attention to this interesting entity. Herein, the authors present their experience with IMT with special emphasis on diagnosis and treatment. METHODS: All records of children treated with diagnosis of IMT between 1977 and 1999 inclusive were evaluated retrospectively. RESULTS: Seven children were treated for IMT with the mean age of 9.14 +/- 2 years (range, 6 to 12 years). Male to female ratio was 5:2. Respiratory symptoms and clubbing were present in a patient with pulmonary IMT (n = 1). Abdominal pain (n = 3), fever (n = 2), and weight loss (n = 4) were encountered in intraabdominal IMTs. The most frequent physical finding was palpable intraabdominal mass (n = 4). Plain films showed nonspecific findings such as radiodense area in the hemithorax (n = 1), displacement of bowel segments (n = 2), air-fluid levels (n = 1), and amorphous calcification (n = 4). Ultrasonography and CT showed calcified masses in 4 cases. Except the case with intrathoracic IMT, all the tumors were located in the abdomen at various sites such as cardioesophageal junction (n = 1), left hepatic lobe (n = 1), mesentery of the small bowel (n = 2), and antimesenteric wall of the descending colon (n = 1), gastrosplenic region and porta hepatis (n = 1). Tumor sizes ranged from 3 x 2 x 2 cm to 15 x 15 x 13 cm. The masses were excised totally in all but one case. Infiltrated organs (esophagogastric junction, a segment of jejunum, and spleen, stomach wall, and renal capsule) were resected in 3 cases. Total surgical excision of IMT was considered adequate for treatment in 6 cases. One patient with aggressive IMT required further treatments such as immunomodulation and chemotherapy and died of neutropenic sepsis. CONCLUSIONS: IMT is a benign neoplasm rarely presented with malignant features such as local invasiveness, recurrence, distant metastasis, or malignant transformation. IMT can be suspected preoperatively through some hematologic abnormalities and radiologic findings, but precise diagnosis should be made on the basis of histologic findings. Complete surgical resection and close follow-up are all necessary for appropriate treatment to avoid recurrences as well as unnecessary and potentially harmful therapy. The optimal management of locally aggressive and recurrent forms should be decided individually for each patient. J Pediatr Surg 36:908-912. PMID- 11381425 TI - Absorption of intraperitoneal CO2 after laparoscopy in piglets: an experimental study. AB - PURPOSE: The aim of this study was to study the absorption time of CO(2) after laparoscopy in piglets. METHODS: Laparoscopies were performed on 14 6-week-old piglets. The animals' abdomens were x-rayed immediately after closure 1, 1.5, and 2 hours postoperatively, and thereafter daily. RESULTS: All CO(2) was absorbed completely within 1 day. CONCLUSION: CO(2) absorption may follow a similar time course in small children. J Pediatr Surg 36:913-916. PMID- 11381426 TI - Surgical treatment of pulmonary hydatid cysts in children. AB - PURPOSE: Ten years' experience is analyzed in pediatric patients with pulmonary hydatid cysts. METHODS: Between 1990 and 2000, 33 pediatric patients (2 to 15 years of age) with pulmonary hydatid cysts were operated on in Department of Thoracic Surgery in Ankara University School of Medicine. There were 17 girls and 16 boys. RESULTS: Twenty-three cases presented as a solitary lung cyst, whereas the remaining 10 were found to have multiple cysts in one or both lungs. Cystotomy and capitonnage were performed in 25 patients, cystotomy was done in 6 patients, and only 2 patients underwent the wedge resection. The authors used no scoliocidal agent in our approach. There was no operative mortality. Recurrence after operation was seen in one patient. CONCLUSION: Pulmonary hydatid cysts in children can be treated successfully by cystotomy and capitonnage or only cystotomy. J Pediatr Surg 36:917-920. PMID- 11381427 TI - Evaluation of 142 consecutive laparoscopic fundoplications in children: effects of the learning curve and technical choice. AB - PURPOSE: This retrospective study describes the 5-year experience of a single surgeon with 142 consecutive laparoscopic fundoplications in children and analyzes the results in terms of the surgeon's learning curve and the choice of technique. METHODS: The patients were 44 girls (40%) and 98 boys (60%) with an age range of 3 months to 18 years (mean, 5.35 years). Indications for surgery included medically refractory reflux associated with vomiting, pneumopathy, otorhinolaryngologic pathology, failure to thrive, esophagitis, apnea and bradycardia, or anemia. The laparoscopic surgery used 5 trocars of 5 mm or 3 mm, with a 30 degrees telescope. RESULTS: Laparoscopic fundoplication was successful in 139 cases. The authors performed 13 Nissen techniques without short gastric vessel division (SGVD), 47 Toupets without SGVD, 9 Toupets with SGVD, and 70 Nissens with SGVD. Mean operating time was 105 minutes (range, 45 to 300). Mean time was 125 minutes for the first 60 cases and 93 minutes for the other 79 cases). Intraoperative and postoperative complication rates were 0.5% and 2%, respectively, and the complications occurred in the first 60 cases. Mean postoperative hospital stay was 3 days (range, 2 to 14). CONCLUSIONS: The rate of complication caused by the laparoscopic procedure was acceptable, and it decreased with the surgeon's experience. Complications and conversions to open technique occurred only in the beginning of the operator's experience. Dysphagia, epigastric pain, gas bloating, and early reflux recurrences were noted among the first 60 cases but seemed to be consecutive to the fact that the gastric vessels were not divided. J Pediatr Surg 36:921-926. PMID- 11381428 TI - Segmental dilatation of the jejunum resembling prenatal volvulus. AB - A case of prenatally diagnosed intestinal obstruction caused by segmental dilatation of the jejunum is presented. Routine ultrasound scan showed mild polyhydramnios, a dilated small bowel loop, and scoliosis. On repeat ultrasound scan a volvulus was suspected, which led to cesarean section and laparotomy of the newborn, revealing the underlying pathology. Segmental resection and end-to end anastomosis were followed by an uneventful recovery. Segmental dilatation of the intestine should be considered as a cause of prenatally manifested intestinal obstruction. Concomitant anomalies are frequent and may be helpful in making the correct diagnosis. J Pediatr Surg 36:927-929. PMID- 11381430 TI - Pericardial foreign body. AB - The authors describe the case of an inhaled foreign body unusually located in the pericardium. An initial chest skiagram was misinterpreted leading to a negative bronchoscopy. Correct anatomic localization of the foreign body only was established at surgical exploration, despite preoperative computerized tomography scan of the chest. Recognition of the propensity of sharp foreign bodies to migrate is essential in treating these patients. A tracheobronchial foreign body migrating to the pericardium has not been reported previously in the literature. J Pediatr Surg 36:936-938. PMID- 11381429 TI - Enteric nervous system, interstitial cells of cajal, and smooth muscle vacuolization in segmental dilatation of jejunum. AB - BACKGROUND/PURPOSE: The etiology of congenital segmental dilatation (CSD) of bowel remains elusive. Intestitial cell of Cajal plays a role in the pace making of the intestine. Its abnormality has been documented in a variety of conditions of abnormal intestinal motility. The current study attempts to evaluate the roles of intestitial cells of Cajal, enteric nervous system, and smooth muscle in segmental dilatation of small bowel. METHODS: Resected specimen from a neonate with segmental dilatation of jejunum was stained with H&E, Alcian blue, Periodic Acid-Schiff (PAS), and immunostained with S100, Ret, MAP5, and c-kit antibodies using the standard immunohistochemical process. RESULTS: The immunostaining of S100, Ret, MAP5 and c-kit of CSD specimen were positive. Localized vacuolization was, however, detected in the circular smooth muscle of the jejunum. The Alcian blue and PAS staining of the vacuolization were negative. CONCLUSIONS: CSD shows no abnormality in the enteric nervous system and pace makers. Localized vacuolization suggests myopathy to be a contributing factor to the disease. J Pediatr Surg 36:930-935. PMID- 11381431 TI - Mixed foregut cyst associated with esophageal atresia. AB - The authors report an unusual case of a thoracoabdominal foregut malformation with components of bronchogenic, esophageal duplication, and pancreatic enterogenous cysts, that presented in a child with esophageal atresia. J Pediatr Surg 36:939-940. PMID- 11381432 TI - Unusual clinical presentation of a rare case of phyllodes tumor of the breast in an adolescent girl. AB - Phyllodes tumor of the breast is a very rare neoplasm, particularly in adolescent girls and young women. It usually presents as unilateral palpable mass. The authors report the case of an adolescent girl who presented at the Emergency Department with bloody nipple discharge. Although the clinical signs suggested an intraductal tumor, the histologic examination showed a phyllodes tumor. The management and the biologic behavior of this uncommon tumor are discussed with particular regard to the very unusual clinical presentation in this patient. J Pediatr Surg 36:941-943. PMID- 11381433 TI - Congenital giant megaureter: first neonatal case. AB - The authors report the first neonatal presentation of a congenital giant megaureter (CGM), a very rare unilateral urinary anomaly, which is defined as "a ureter whose lumen is congenitally, focally and segmentally dilated to more than 10 times the normal diameter, in presence of normal bladder volume and function." The definition of CGM is purely descriptive; it can be secondary to several different diseases, and in the series of 21 CGM (12 in duplex system) reported by Chen-Ju Huang, the morphology of the distal ureter varied from a near normal caliber to a ureteral stenosis, a ureterocele or a complete ureteral atresia. J Pediatr Surg 36:944-945. PMID- 11381434 TI - Soft tissue infection caused by Kingella kingae in a child. AB - During the last years an increasing number of reports concerning Kingella kingae infections in children has been published. Most cases were osteoarticular infections. The authors report the clinical and laboratory findings from a 3-year old child with a presternal soft tissue infection due to K kingae. After surgical excochleation and antibiotic treatment there was an uneventful recovery. J Pediatr Surg 36:946-947. PMID- 11381435 TI - Surgical implications of pseudomonas aeruginosa necrotizing fasciitis in a child with acute lymphoblastic leukemia. AB - Necrotizing fasciitis caused by Pseudomonas aeruginosa is extremely rare. Only 4 cases were reported in the literature. The authors report the occurrence of P aeruginosa necrotizing fasciitis starting out as a vulval abscess in a girl before induction chemotherapy for acute lymphoblastic leukemia. To our knowledge, this is the second case described in association with leukemia. In this case, the outcome was favorable because of early surgical intervention, confirming the diagnosis. J Pediatr Surg 36:948-950. PMID- 11381436 TI - Intraoperative endoscopy in a child with Turner's syndrome and gastrointestinal hemorrhage: a case report. AB - Gastrointestinal bleeding in Turner's syndrome can represent vascular lesions that are frequently beyond standard endoscopic reach and often life threatening. This report describes the successful use of intraoperative endoscopy to identify the souce of bleeding in an adolescent with Turner's syndrome and significant intestinal hemorrhage. J Pediatr Surg 36:951-952. PMID- 11381437 TI - Late recurrence of neuroblastoma stage 4S with unusual clinicopathologic findings. AB - The authors present unusual clinicopathologic findings of a patient with neuroblastoma stage 4S that recurred 11 years after induction of complete remission with chemotherapy. A 12-year-old girl presented with recurrent tumor in the liver. Urinary catecholamine metabolites were within normal range in contrast to the increased values at initial presentation. She underwent left lateral segmentectomy and biopsy of the right lobe. Histologic analysis of the recurrent tumor showed undifferentiated neuroblastoma intermingled with mature ganglioneuromatous lesions. There also were scattered ganglioneuromatous lesions throughout nontumorous area of the liver. Although multimodal intensified treatments including autologous bone marrow transplantation were performed, the patient died of obstinate recurrent tumor at age 14 years. The clinicopathologic findings suggested dedifferentiation from the ganglioneuromatous lesion rather than ordinary recurrence of the primary tumor. The current case and the literature review may indicate that long-term follow-up would be necessary for neuroblastoma stage 4/4S cases. J Pediatr Surg 36:953-955. PMID- 11381438 TI - Duodenal atresia with an anomalous common bile duct masquerading as a midgut volvulus. AB - In a patient with duodenal atresia, a "double bubble" is classically present on plain radiographs. When bowel gas exists distal to the duodenum, duodenal atresia often is excluded from the differential diagnosis. The authors present a case in which contrast can be seen in the small bowel and biliary system on upper gastrointestinal series in a patient with duodenal atresia and an anomalous common bile duct. One always must consider duodenal atresia with an anomalous biliary system as a possible cause of bilious vomiting with a high grade proximal bowel obstruction in a neonate. J Pediatr Surg 36:956-957. PMID- 11381439 TI - H-type tracheoesophageal fistula. PMID- 11381440 TI - Quality of life for children with fecal incontinence after surgically corrected anorectal malformation. PMID- 11381442 TI - Ureterocystoplasty: an alternative reconstructive procedure to enterocystoplasty in suitable cases. PMID- 11381444 TI - Chyloperitoneum: a postoperative complication after repair of tracheoesophageal fistula. AB - Chyloperitoneum is rare condition that can occur in response to various pathologic process. The authors described a newborn baby who had chyloperitoneum after primary surgical repair of esophageal atresia with tracheoesophageal fistula (Gross type C). This probably resulted from iatrogenic damage of the thoracic duct during ligation of azygous vein resulting in leaking lacteal within the intestinal wall. Biochemical analysis of ascitic fluid, plasma, and milk formula for triglyceride and cholesterol level can differentiate neonatal gastric intestinal perforation. PMID- 11381445 TI - Mammary duct ectasia in children presenting bloody nipple discharge: a case in a pubertal girl. AB - Mammary duct ectasia occurs rarely in childhood. The authors report on the case of a pubertal girl who was operated on for duct ectasia with bloody nipple discharge. Duct ectasia is regarded as a primary lesion; it is considered to be a cause of bloody secretion, and it has a mechanism similar to that of mammary duct papilloma. PMID- 11381446 TI - Use of the gonadal vein in children with difficult central venous access: a novel technique. AB - In children who require multiple central venous catheterization, the commonly used veins can become thrombosed making central venous access a challenging problem. The authors report on a patient with a rare metabolic disorder who had extensive thrombosis of the superior vena cava and iliofemoral veins, in whom the right ovarian vein was cannulated using a retroperitoneal approach for the placement of a SILASTIC((R)) (Dow Corning, Midland, MI) catheter with an implantable port. This is a useful and reliable technique in children who have difficult venous access. PMID- 11381447 TI - Congenital cystic adenomatoid malformation of the lung with an esophageal cyst: report of a case. AB - The authors report the case of a 7-month-old girl found to have both congenital cystic adenomatoid malformation of the lung (CCAM) and esophageal cyst. She suffered repeated episodes of pneumonia and exhibited signs of respiratory distress on admission to our hospital. Chest radiography and magnetic resonance imaging (MRI) showed 2 different kinds of cystic lesions. Resection of the lower lobe of the right lung and excision of the posterior mediastinal cyst were performed. Histologic examination showed Stocker type I CCAM and esophageal cyst. Coexistence of both CCAM and esophageal cyst is extremely rare. The authors speculate that the pathologies of this case originated from a regional disturbance of common embryologic origin during 2 different phases of lung-bud foregut malformations. PMID- 11381448 TI - Presentation of carcinoma in a patient with a previous operation for Hirschsprung's disease. AB - The usual complications after a definitive pull-through procedure for Hirschsprung's disease include stricture formation, enterocolitis, bowel obstruction, and, occasionally, wound infection. The authors report a case of mucinous adenocarcinoma arising 32 years later at a stricture site from a previous pull-through procedure. PMID- 11381449 TI - Mucinous cystadenoma of the ovary in a 15-year-old girl. AB - Benign neoplasms of the ovary originating from epithelial tissue are common tumors in adult women. They are, however, rarely seen in children or adolescent girls. Here the authors present a case of an ovarian mucinous cystadenoma in a premenarchal girl. To our knowledge, there are only 5 other cases reported in the literature. PMID- 11381450 TI - Accessory liver in an infant with congenital diaphragmatic hernia. AB - Incidence of congenital diaphragmatic hernia (CDH) ranges from 1 in 2,400 to one in 5,000 live births. Associated anomalies in CDH are approximately 30% to 35%. The authors report on an infant with CDH and an accessory supradiaphragmatic liver with separate vascularization from the thoracic aorta. The pathogenesis of this malformation is discussed. The postoperative course and the follow-up of the child over 5 years was completely uneventful. PMID- 11381451 TI - Laparoscopic excision of subdiaphragmatic epidermoid cyst: a case report. AB - Retroperitoneal epidermoid cysts are rare. The authors report a case of an 11 year-old boy with an asymptomatic subdiaphragmatic cyst, which was found incidentally during an investigation for hypertension. At laparoscopy, the cyst was densely adherent to the diaphragm, resulting in a pneumothorax during dissection. Nevertheless, the excision and the diaphragmatic repair could be completed laparoscopically without complication. Microscopic examination showed an epidermoid cyst. No similar case has been reported in the literature. PMID- 11381452 TI - Use and abuse of antimicrobials in periodontal therapy. PMID- 11381453 TI - Altered passive eruption. AB - Biologic contributions to the smile play an important role in esthetics. The impact of the gingival complex on the esthetic qualities of a smile are significant, and both patients and dentists are developing a greater appreciation for the impact of gingiva on the beauty of a smile. Historically, the focus of periodontal therapy has been on managing patients with longer-appearing teeth secondary to attachment loss through pathology or as a result of periodontal surgery. Nevertheless, early diagnosis and treatment, as well as modern techniques and materials, have helped periodontology evolve to a point where such problems occur less frequently. PMID- 11381454 TI - Barriers to sustainable water-quality management. AB - Due to the pressures of increasing population and developing economy all over the world, the present situation of water-quality management is far from satisfactory. To enhance sustainability of water-quality-management systems, in depth research of the related barriers and the relevant mitigation approaches is desired. In this paper, recent developments, advancements, challenges, and barriers associated with practices of water-quality management were analyzed. A number of related methodologies, applications, and policy considerations were examined. Issues of information support, technology development, system integration, and policy implementation were discussed. Perspectives of sustainable water-quality management in the twenty-first century were investigated, demonstrating many demanding areas for enhanced research efforts, including issues of data availability and reliability, concerns in system complexity and methodology validity, limitations of computer techniques, usefulness of research outputs, difficulties in policy implementation, and necessity of training programs. PMID- 11381455 TI - A two-dimensional water-quality model for a winding and topographically complicated river. AB - In this paper, a two-dimensional numerical calculation algorithm for water quality modeling is presented. The algorithm is designed specifically for river systems with complicated geometric conditions. When velocity field data of the river are not available, the numerical calculation algorithm for the water quality modeling can be used to project river-water quality by using a topographic map of the river course and a finite element method. The calculation results of the water-quality model can show the concentration fields of various pollutants. The water-quality model was applied to a case-study in the Hengyang City section of Xiangjiang River in Hunan Province, China. The river under consideration is winding and has an isle between two branches. In 1995, Chinese government secured a World Bank loan to conduct a Waterways Project in the study region. It was expected that construction works in the river section might affect water quality. Given that the project would change the hydrological regime of the river system and discharges, and so would affect water quality, there would be a need for model results that would predict the water-quality impacts of the Waterways Project. In particular, the study intended to apply the model to identify changes in river-water quality associated with the construction of Dayuandu navigation key project. It is hoped that the numerical calculation algorithm for the water-quality modeling presented in this paper can also be applied to other shallow rivers with similar topographical conditions. PMID- 11381456 TI - Integrating water-quality management and land-use planning in a watershed context. AB - The spatial relationships between land uses and river-water quality measured with biological, water chemistry, and habitat indicators were analyzed in the Little Miami River watershed, OH, USA. Data obtained from various federal and state agencies were integrated with Geographic Information System spatial analysis functions. After statistically analyzing the spatial patterns of the water quality in receiving rivers and land uses and other point pollution sources in the watershed, the results showed that the water biotic quality did not degrade significantly below wastewater treatment plants. However, significantly lower water quality was found in areas downstream from high human impact areas where urban land was dominated or near point pollution sources. The study exhibits the importance of integrating water-quality management and land-use planning. Planners and policy-makers at different levels should bring stakeholders together, based on the understanding of land-water relationship in a watershed, to prevent pollution from happening and to plan for a sustainable future. PMID- 11381457 TI - Total waste-load control and allocation based on input-output analysis for Shenzhen, south China. AB - The general objective for this paper is to reveal the dynamic relationships between the rapid economic development, water pollution and the subsequent waste load allocation in different economic sectors through a case-study in Shenzhen City, South China. Two-objective analysis model was employed based on the input output table for Shenzhen with the full consideration of various constraints in local area. The improved Tchebycheff procedure was used for obtaining the solutions. The predictions were made on economic development and pollutants from wastewater in different sectors and different planning years. The present study allows for the consideration of the economic structural adjustment. It is found that the current situation of economic structure is generally good and is subject to further adjustment in Shenzhen, although it has undergone the rapid development in the past 18 years. When the maximum Gross Domestic Production and the minimum Chemical Oxygen Demand are chosen as the two objectives subject to other constraints, the harmonized results indicated a scheme that claims substantial reduction of polluting effluences in Shenzhen while closely keeping the economic growth rate as planned. PMID- 11381458 TI - Integrated management of radioactive strontium contamination in aqueous stream systems. AB - A combination of biomass treatment, fluidized bed/membrane reactor, and a minimum suspension fluidized bed reactor is proposed to remove strontium cations from aqueous solutions, such as those generated by nuclear reactors. After conducting a series of screening tests, three adsorbents were selected for their suitability and high adsorptive capacity. The proposed combination uses Chlorella vulgaris in a packed column, followed by the fluidized bed/membrane reactor with bentonite powder in suspension. The membrane is primarily used to retain bentonite powder in the reactor. However, the same can be designed to remove additional amount of contaminant from the aqueous stream. The final separation is carried out in a fluidized bed containing resins that are suspended with minimal airflow. In laboratory scale, a flow rate of 600 ml/h was achieved for 30 min during which period the inlet concentration of 100 mg/l was reduced to 2.5 mg/l at the outlet. Bio-encapsulation with thermophilic bacteria and subsequent separation is proposed at this point in order to reduce the concentration to an even lower level. The proposed separation scheme offers an acceptable solution to removing strontium while minimizing the generation of secondary waste. PMID- 11381459 TI - Assessing pollution prevention program by QUAL2E simulation analysis for the Kao Ping River Basin, Taiwan. AB - Wise and sustainable uses of water resources are essential for an effective river basin-management planning. Recent management strategy further addresses the fact that quantity and quality of water are closely interrelated, and both must be considered simultaneously for all water resources and water quality management practices. The aim of this paper is to explore the impacts of water resources redistribution and pollution prevention actions between and within river basins simultaneously in South Taiwan. Much emphasis will be placed on assessing the impacts of water transfer over natural boundary to satisfy the needs of industrial development in the Tseng-Wen River system and its resultant influence on the water quality in the downstream area of the Kao-Ping River system where the pollution prevention program is to be implemented. The Kao-Ping River was further characterized hydraulically and environmentally, based on a full investigation of discharges and withdrawals in the river reaches. QUAL2E was successfully calibrated and validated using data collected between 1998 and 1999, and the model was capable of predicting the concentrations of biochemical oxygen demand, dissolved oxygen, total phosphate-phosphorus, and ammonia-nitrogen (NH3 N) for the entire river system. With the aid of QUAL2E simulation model, it shows eliminating the pig farming activities and constructing the sewer systems in the upstream area of Kao-Ping River cannot guarantee the full compliance with water quality standards in the downstream area and water transfer in the upstream area further increases negative impacts on the water quality in the wet season. The predicted situation of water quality in the dry season may even present worse condition. Additional water pollution control policy, such as the use of economic instruments, for controlling and reducing the waste-load of biochemical oxygen demand and ammonia-nitrogen is needed in the Kao-Ping River system in the long run. PMID- 11381460 TI - Simulation of cooling-water discharges from power plants. AB - Accurate simulation of the temperature distribution in a cooling lake or reservoir is often required for feasibility studies of engineering options that increase the cooling capacity of the waterbody. A three-dimensional hydrodynamic and temperature model has been developed and applied to several cooling lakes in the south-eastern United States. In this paper, the details of the modeling system are presented, along with the application to the Flint Creek Lake. PMID- 11381461 TI - A system dynamics approach for regional environmental planning and management: a study for the Lake Erhai Basin. AB - In recent years, water-quality deterioration associated with rapid socio-economic development in the Lake Erhai Basin, China, has acquired more and more attention from the public and the government. An effective planning for the basin's environmental management system is desired for sustainable regional development. In this study, an environmental system dynamics model, named ErhaiSD, is developed for supporting this planning task. The ErhaiSD consists of dynamic simulation models that explicitly consider information feedback that governs interactions in the system. Such models are capable of synthesizing component level knowledge into system behaviour simulation at an integrated level. This capability is very useful in analyzing and recommending policy decisions. For the study case, interactions among a umber of system components within a time frame of 15 years are examined dynamically. Four planning alternatives are considered. The base run is based on an assumption that the existing pattern of human activities will prevail in the entire planning horizon, and the other alternatives are based on previous planning studies. The contributions of various nonpoint pollution sources to the lake's eutrophication problems, and the effects of industrial activities and wastewater treatment processes on pollution problems in the Xier River are analyzed through the developed modeling system. The exercise draws attention to the implications of different alternatives to the system's environmental and socio-economic objectives. The modeling results are directly useful for simulating and evaluating a variety of decision actions and their dynamic consequences, and answering questions such as 'What should I do?', 'What if I do?' and 'What are the expected consequences?'. PMID- 11381462 TI - [Pharmacokinetics and tissue distribution of ipecac syrup]. PMID- 11381463 TI - [22nd Annual meeting Japanese Society for Clinical Toxicology Panel discussion. Reassessment of the initial management of poisoned patients]. PMID- 11381464 TI - [Acute isoniazid poisoning presenting convulsion and liver dysfunction]. AB - A-19-year-old girl ingested 7.4 g of Isoniazid (INH) and 14.5 g of Ethambutol (EB) intentionally. She presented repetitive generalized convulsions with metabolic acidosis, treated with diazepam and 7.5 g of Pyridoxine (Vitamin B6). During day 4 and 5, liver dysfunction worsened and prothrombin time declined below 5%. She was treated with infusion of fresh frozen plasma and fully recovered on day 13. Serum concentration of INH and EB at 5 hours after ingestion was 7.6, 0.94 micrograms/ml respectively. PMID- 11381465 TI - [Gut decontamination--2]. PMID- 11381466 TI - [Acute poisoning induced by nitrogen dioxide inhalation in Japan]. PMID- 11381467 TI - Protective headgear for midwestern agriculture: a limited wear study. AB - Baseball caps are popular with farm workers, but have been criticized because they do not sufficiently shade the face, neck, and ears. U.S. Environmental Protection Agency standards require workers to wear chemical-resistant hoods or chemical-resistant hats with wide brims during the application of pesticides whose labels call for head protection. In this study, four farm workers wore baseball caps and two alternative types of headgear with wide brims for 20 to 36 hours during planting of corn and soybeans to compare performance features and practicality. Afterwards, researchers analyzed the headgear fabrics by gas chromatography or high-performance liquid chromatography to determine the levels at which five herbicides were deposited on the headgear: 2,4-D, metolachlor, acetochlor, ethalfluralin, and glyphosate. Chemical analysis revealed that 12 percent of specimens had detectable residue: levels of glyphosate in the nanograms-per-square-centimeter (ng/cm2) range and levels of 2,4-D in the micrograms-per-square-centimeter (microgram/cm2) range. Workers, however, preferred the baseball caps because of problems with the wind and feelings of embarrassment about wearing other types of headgear. An acceptable, protective substitute for the baseball cap has yet to be designed. PMID- 11381468 TI - Scientific writing as an art and as a science. PMID- 11381469 TI - Undergraduate environmental health education: preparing for the future. AB - Attendees indicated that the workshop was beneficial and that the opportunity to communicate with faculty representing 23 programs accredited by EHAC and nine programs not accredited by EHAC was extremely useful. There was general agreement on a number of points: There is a need for undergraduate environmental health programs to collaborate on matters related to distance learning. Topics related to women, gender, diversity, ethics, and international environmental health should be incorporated into the environmental health curriculum. There are no major problems with current EHAC curricular guidelines, but the guidelines should be evaluated on a regular basis. Field experience or internship is an essential component in the academic preparation of undergraduate environmental health students. There is a significant need for increased funding for undergraduate environmental health programs. There is a need to increase the visibility and recognition of environmental health programs. There is a need to solidify ties with traditional partners and to establish new linkages at the local, regional, and national levels in the government, community, and private sector. It is essential that undergraduate faculty communicate with each other on matters relating to the preparation of environment health practitioners. There is a need for an association of undergraduate academic programs to provide leadership and a focal point for identification and resolution of issues common to all. The establishment of an association was viewed as the most practical and effective way to address these issues and to pursue related opportunities. PMID- 11381470 TI - Nontypical Bacillus cereus outbreak in a child care center. AB - On October 29, 1998, The Garland Health Department in Texas investigated seven illnesses at a church day school. The six children and one adult had handled hydrated, orange-colored rice before consuming a meal. B. cereus organisms were found in the rice at an estimated concentration of 5.6 x 10(5) per gram. As far as the authors can determine, this outbreak is the first one documented to have occurred from indirect ingestion of B. cereus by way of contaminated hands, as opposed to ingestion of the toxin in the food product in which the organisms grew. PMID- 11381471 TI - On the path from student to practitioner: testing the water before the internship. AB - Individuals entering careers in environmental health or safety must either decide to be generalists or must specialize in one of several diverse areas. At Illinois State University, the Environmental Health Program and the Safety Program coordinate one-day student-practitioner partnerships that provide students with the opportunity to observe professionals in the field. These partnerships are called "Professional Days" and help students experience a potential carrier before beginning an internship. The authors discuss how the visits are coordinated, what role workplace visits play in students' selection of career paths, and how both students and employers evaluate the experience. PMID- 11381472 TI - Bioterrorism. AB - Although biological agents have been used in warfare for centuries, several events in the past decade have raised concerns that they could be used for terrorism. Revelations about the sophisticated biological-weapons programs of the former Soviet Union and Iraq have heightened concern that countries with offensive-research programs, including those that sponsor international terrorism, might assist in the proliferation of agents, culturing capability, and dissemination techniques, and might benefit in these undertakings from the availability of skilled laboratory technicians. Release of sarin nerve agent in the Tokyo subway system in 1995 by the Aum Shinrikyo cult demonstrated that in the future terrorists might select unconventional weapons. Certain properties of biological pathogens may make them the ideal terrorist weapon, including 1) ease of procurement, 2) simplicity of production in large quantities at minimal expense, 3) ease of dissemination with low technology, and 4) potential to overwhelm the medical system with large numbers of casualties. Dissemination of a biological agent would be silent, and the incubation period allows a perpetrator to escape to great distances from the area of release before the first ill persons seek medical care. Countermeasures include intelligence gathering, physical protection, and detection systems. Medical countermeasures include laboratory diagnostics, vaccines, and medications for prophylaxis and treatment. Public health, medical, and environmental health personnel need to have a heightened awareness, through education, about the threat from biological agents. PMID- 11381473 TI - Investigation of illness associated with exposure to hydrogen sulfide among Pennsylvania school students. AB - During 1998, the Pennsylvania Department of Health received complaints about hydrogen sulfide odors believed to be associated with mushroom-composting operations in southeastern Pennsylvania. Many residents were concerned about possible illness in students attending an elementary school near the composting operations. In response, the department conducted health surveys during the spring and autumn at the exposed school and at a nearby control school. The surveys assessed whether exposures to hydrogen sulfide were associated with excess adverse health effects by comparing health effects among students from the exposed school with those among students from the control school. School nurses were trained to complete health questionnaires for the students. The state environmental agency measured daily ambient hydrogen sulfide concentrations at both schools. No consistent association was found between exposure to low levels of hydrogen sulfide and any adverse health effects. It was concluded that the students attending the elementary school near the mushroom-composting operations were not exposed to any significant public health hazard. PMID- 11381474 TI - Reducing the stigma of schizophrenia: understanding the process and options for interventions. PMID- 11381475 TI - Combating the stigma of schizophrenia. PMID- 11381476 TI - Epidemiology in community psychiatric research: common uses and methodological issues. AB - OBJECTIVE: Epidemiological principles underpin much medical research particularly that concerned with the planning and evaluation of health services, including research in community and social psychiatry. The aim of this paper is to review some of the common uses of epidemiology in community psychiatric research and to discuss some methodological issues that arise frequently in epidemiological research in community settings. METHODS: This is a review of the relevant literature and of the work currently in progress in the department of psychological medicine of the university of Wales College of Medicine. RESULTS: Among the many uses of epidemiology in health care, four are especially relevant in community psychiatric settings: the assessment of the mental health needs of the population (four approaches are described: the collection of routine data, surveys of existing patients, surveys of the general population and statistical modelling), the identification of risk factors of disease, the contribution to prevention and the assessment of the clinical effectiveness of health care interventions. The most important methodological issues include causal inference which in epidemiology takes the form of explaining the association between an exposure and disease (chance, bias, confounding, reverse causality and causality), the issue of confounding and how to adjust for it and issues arising in the context of specific study designs. CONCLUSION: Epidemiology has become a set of methods used to investigate a wide range of clinical questions. Population based research is an essential part of clinical research but epidemiological knowledge is also needed by clinicians in order to critically appraise and interpret the scientific literature. PMID- 11381477 TI - [Group cognitive behavior therapy in the routine care at a Psychiatric Ward of Diagnosis and Treatment]. AB - OBJECTIVE: To describe a preliminary evaluation of a group CBT approach which has been fully integrated into the routine care for acute psychotic inpatients, admitted to a typically overburdened inner-city psychiatric general hospital unit. DESIGN: The design is pre-post. A comparison of some indicators between the years preceding (1997) and following the beginning of the group therapy (1998) has been carried out. SETTING: Psychiatric ward of S. Filippo Neri in Rome, in 1998. METHOD: The study compared the rate of rehospitalization, the rate of compulsory admission, the use of physical restraints, the episodes of violent behaviour and the escape from the ward. During the last three months of 1998 patients' satisfaction was also measured. RESULTS: Overall 385 patients attended the group therapy, 79% of all inpatients in the period, with an average number of 4 sessions per patient. 59% of participants in the group in 1998 had a diagnosis of schizophrenia or paranoid disorder. The total compulsory admissions declined by 1/3 (p < 0.001), the "revolving door" patients declined by 1/3 (p < 0.05), compulsory admissions in "revolving door" patients declined by 3/4 (p < 0.005), violent episodes by almost half (p < 0.001), physical restraints and patients escape from the ward almost disappeared. Patients' opinion about the participation to the group was excellent. CONCLUSIONS: It is likely, but not sure, that the improvements were directly attributable to the introduction of the group treatment. Moreover, the group was clearly found to be useful by the majority of the participants. Some asked for further help of this type on discharge. These preliminary results are promising and make us confident they will act as stimuli for other colleagues to implement and evaluate this approach in other settings. PMID- 11381478 TI - [Decision making process for compulsory admission: study of a group of psychiatrists of Sardinia, Italy]. AB - OBJECTIVE: The aim of the study was to investigate the attitudes in the decision making process for admitting patients compulsorily under the Italian Law 180/78. SETTING: A group of psychiatrists (n = 81) working in the area of Cagliari (Sardinia, Italy). MAIN OUTCOME MEASURES: Three case-vignettes describing different hypothetical clinical situations: 1) a man with depression and psychotic symptoms; 2) a woman with a possible first episode of psychosis; and 3) a man with a history of substance abuse and bipolar disorder. Each vignette was followed by a list of 11 factors reported in the literature as important in the decision to admit compulsorily (current mental state, severity of the disease, dangerousness to self or others, psychiatric history, likely response of the mental state to the medical treatment, age and gender of the patient, owning a home, occupational status, social support available). Psychiatrists were asked: a) whether they would admit the patient compulsorily; and b) to rate 11 factors from the most to the least important in their decision. RESULTS: We obtained responses from 57 psychiatrists (26 males and 31 females). The most important factors for deciding to detain a patient compulsorily were: current mental state, diagnosis, severity of the illness and possible contribution of medical treatment. Perceived dangerousness to self and others was considered the most important factor by 23% of the sample. CONCLUSIONS: Our results confirm that, in three different hypothetical situations, there is agreement in the factors perceived as the most important in the decision to admit compulsorily, although their application is highly subjective. The factors correspond to those recommended by the Italian Law 180. The need for protection of the patient or others has an important role in the decision and may take precedence over the current mental state of the patients [corrected]. PMID- 11381479 TI - Depression and social change. From transcultural psychiatry to a constructivist model. AB - Based on the findings of previous studies carried out by our group, which will be briefly summarised, the present paper puts forward several hypotheses to account for the evolution of depressive symptoms and the possible increase observed in risk of depression subsequent to social changes. The particular mood disorders presented by Senegalese emigrants and several protective factors which appear to determine a low risk in these populations, such as a strong social support, will be examined. Based on a previous investigation carried out by our group, which seems to indicate the presence of depressive pictures among poorly westernised populations such as the Peul nomads or Dogon farmers from the Sub-Saharian regions, the hypothesis that "westernalisation" (considered as the loss at an individual level of traditional ways of life, working habits, cultural patterns and languages in favour of different attitudes influenced by western culture) may represent a risk factor for depressive illness, in its clinical expressions commonly observed in western contexts, has been considered. In these populations, with the exception of educated individuals, the albeit rare depressive symptoms appear to be secondary to serious somatic disorders. Research performed identified two well-defined means of clinical expression, which has been termed, respectively, "western style" or "guilty" and "traditional" or "dislocation from the group". Further studies carried out in rapidly changing areas seem to indicate how environmental factors are able to influence the evolution of depressive symptoms from the first form to the latter and to modify the threshold of onset of emotive, behavioural and depressive patterns. It has been hypothesised that rapid changes in the social organisation tend to exacerbate attitudes of "compulsive hyper-responsibilisation", a cognitive set of basic assumptions which may be considered at the same time both as a product of "westernalization" at an individual level and a risk factor for depression. Individuals who possess these basic characteristics, subsequent to the opportunities afforded by the social changes, tend to develop new complex systems of interpreting reality, causality, controlling of events and ways of expressing emotions. Accordingly, we herewith propose a reviewal of the entire threshold concept and provide a means of interpreting the transformation in depressive phenomena in view of the fact that, although the new levels of knowledge and learning better equip subjects to face the new situations, they also render them more vulnerable to depression. PMID- 11381480 TI - Stigma: many mechanisms require multifaceted responses. PMID- 11381481 TI - The child with persistent cough. AB - Coughing is a healthy reflex. Causes of a cough can vary from minor upper respiratory illnesses to malignancy. When a child's cough continues for weeks, parents worry. Primary care providers must decide when reassessment is needed and if a vigorous workup and referral to a pulmonologist are required. The above discussion should assist these physicians. PMID- 11381482 TI - When anthrax was a Delaware Valley disease: Dr. Herman Gold's experience with cutaneous anthrax. PMID- 11381483 TI - Physician leaders hear details on pharmacist collaborative drug therapy management. PMID- 11381484 TI - Computerized prescriber order entry changes pharmacists' roles. PMID- 11381485 TI - Gender gap narrows in smoking prevalence, deaths. PMID- 11381486 TI - Noninvasive glucose-monitoring device gains FDA approval. PMID- 11381487 TI - CDC reports higher levels of other phthalates than of DEHP in humans, despite greater environmental exposure. PMID- 11381488 TI - JCAHO selects elements for random unannounced surveys. PMID- 11381489 TI - Drug-use principles released by USP. PMID- 11381490 TI - Health care: a business or a responsibility? PMID- 11381491 TI - Providing pharmaceutical care for indigent patients: a roundtable discussion. PMID- 11381492 TI - Thymosin alpha-1. AB - The pharmacology, pharmacokinetics, clinical efficacy, adverse effects, and dosage and administration of thymosin alpha-1 (TA1) are reviewed. TA1 is a synthetic polypeptide. The drug is in Phase III trials for the treatment of hepatitis C and in Phase II trials for hepatitis B. Additional possible indications are malignant melanoma, hepatocellular carcinoma, drug-resistant tuberculosis, and DiGeorge's syndrome. TA1 is thought to modulate the immune system by augmenting T-cell function. TA1 may affect thymocytes by stimulating their differentiation or by converting them to active T cells. TA1 is rapidly absorbed, achieving peak serum concentrations within two hours. Blood levels return to baseline within 24 hours, and the serum half-life is approximately 2 hours. TA1's efficacy in hepatitis B has been evaluated in 195 patients in four clinical trials. One study found hepatitis B virus (HBV) DNA clearance at six months in 9 of 17 patients receiving TA1, compared with 10 of 16 patients treated with interferon alfa-2b (IFN-alpha 2b) and 4 of 15 historical controls. An open label trial found HBV DNA clearance in 53% of patients at six months. A randomized, controlled trial found HBV DNA clearance in 40.6% and 25.6% of patients treated with TA1 for 6 and 12 months, respectively, compared with 9.4% of untreated controls. Efficacy for hepatitis C has been evaluated in 162 patients in three clinical trials. In one trial, the number of patients who achieved normal serum alanine aminotransferase (ALT) levels did not differ significantly between TA1 and placebo. In the other two trials, combination TA1 and IFN-alpha 2b was compared with IFN-alpha 2b alone. One trial found a normal serum ALT level at six months in 71% of patients receiving combination therapy, versus 35% of patients receiving IFN-alpha 2b alone. Hepatitis C virus RNA clearance occurred in 65% of patients treated with combination therapy and 29% of patients treated with IFN-alpha 2b alone. The third trial, comparing combination TA1 and IFN-alpha 2b with IFN-alpha 2b alone and with placebo, found normalization of ALT levels at six months in 37.1% of patients receiving combination therapy, 16.2% of patients receiving IFN-alpha 2b alone, and 2.7% of patients receiving placebo. TA1 is well tolerated. Most studies observed only local irritation at the injection site. For hepatitis B and C, TA1 1.6 mg (900 micrograms/m2) should be administered subcutaneously twice a week. Clinical trials of TA1 for chronic hepatitis B or C have had mixed results. TA1 may be useful as monotherapy for hepatitis B or in combination with IFN-alpha 2b for hepatitis C, but its effects on morbidity and mortality remain to be seen. PMID- 11381493 TI - Perceived value and outcomes of residency projects. AB - Residency program directors' attitudes toward residency projects were studied. A questionnaire about the residency project experience was mailed in January 2000 to 446 pharmacy practice residency program and specialty residency program directors in the program database of the American Society of Health-System Pharmacists. Recipients responded to opinion statements on a 5-point scale. Responses to the opinion statements were separated into seven categories for analysis. A total of 278 usable questionnaires were returned, for a raw response rate of 63.6%. During the preceding three years, residency directors had served as primary advisors on 917 projects; 171 had served as advisor on at least one of every type of project allowed in the accreditation standards. Of the 917 projects, 364 were presented at national professional meetings, 124 were published, and 484 were believed to have resulted in a positive change in pharmacy services. There were no significant differences in total response scores among any of the subgroups analyzed. There was strong agreement that residency projects were valuable and should continue to be part of the residency program experience. The directors' views of the importance of original research as a project option were more neutral. Overall residency program directors had positive perceptions of the value of residency projects to both residents and institutions and believed that they should continue to be a requirement of residency programs. PMID- 11381494 TI - Certificate program in teaching for pharmacy residents. PMID- 11381495 TI - Therapeutic interchange of fluoroquinolones at a medical center. PMID- 11381496 TI - Medical errors: creating the tension for change. PMID- 11381497 TI - Prolonged shutdown of an isolator because of false-positive sterility tests. PMID- 11381498 TI - Filtration of radiopharmaceuticals. PMID- 11381499 TI - Expression of cathepsin B and cystatin C in human breast cancer. AB - Cathepsin B, which was originally found to be a lysosomal cysteine protease, is also an important matrix protease. In this study, we investigated the expression of cathepsin B and cystatin C, the strongest inhibitor of cathepsin B, and measured the relative amounts of each in human breast cancer tissues. Cystatin C expression relative to cathepsin B expression was found to be decreased. This finding could be associated with the looseness of cancerous interstitial tissue, which might play a role in cancer invasion and metastasis. This report documents the first simultaneous investigation of cathepsin B and cystatin C in breast cancer tissues. PMID- 11381500 TI - Dissociation between T helper type 1 and type 2 differentiation and cytokine production in tumor-infiltrating lymphocytes in patients with lung cancer. AB - We recently reported the balance of T helper type 1 (Th1) cells and type 2 (Th2) cells in patients with lung carcinomas. This study was conducted to investigate their activity and role in tumors, which remain unclear. We determined the population of lymphocytes with intracellular interferon (IFN)-gamma or interleukin (IL)-4 by flow cytometry, and investigated cytokine production using enzyme-linked immunosorbent assay (ELISA) in 22 patients with non-small cell lung cancer. The IFN-gamma-positive subset showed a significant increase in the number of tumor-infiltrating lymphocytes (TIL) compared with the peripheral blood lymphocytes (PBL) (PBL, 13.8% +/- 1.5%; TIL, 34.3% +/- 3.4%; P < 0.001), and the IL-4 positive subset showed reverse results (PBL, 3.7% +/- 0.6%; TIL, 2.1% +/- 0.3%; P = 0.037). However, TIL did not produce more IFN-gamma than PBL. The results of intracellular IFN-gamma analysis and the production of IFN-gamma in PBL and TIL were significantly correlated (PBL: n = 22, r = 0.50, P = 0.025; TIL: n = 22, r = 0.44, P = 0.022). The dissociation between Th1 differentiation and IFN-gamma production in TIL was one of the host factors influencing the immune anergy against tumors. PMID- 11381501 TI - Early removal of chest drainage tubes and oxygen support after a lobectomy for lung cancer facilitates earlier recovery of the 6-minute walking distance. AB - The aim of this study was to assess the effects of the early removal of chest tubes and oxygen support lines on the postoperative recovery of patients, who underwent a lobectomy for lung cancer. Forty-two patients, in whom the removal of chest tubes and oxygen support lines was planned for the morning after surgery (subjective group), were matched by sex and age with 42 patients for whom no such action was scheduled (control group). The mean duration of chest tube drainage was 1.5 +/- 0.8 days in the subjective group, which was significantly shorter than the period of 2.8 +/- 1.0 days in the control group (P < 0.001). The mean duration of oxygen support was 1.1 +/- 0.3 days in the subjective group, which was significantly shorter than the period of 3.1 +/- 1.3 days in the control group (P < 0.001). There was no significant difference in the chest drainage volume and oxygen saturation on the morning after surgery between the two groups. We thus compared the postoperative changes in vital capacity (VC) and 6-min walking distance (6MWD) after surgery between the two groups. The early removal of chest tubes and oxygen support lines significantly reduced the impairments of 6MWD 1 week after surgery (P = 0.04) and also diminished the impairments of VC 1 week after surgery but not to a significant extent (P = 0.06). The early removal of chest tubes and oxygen support lines could accelerate the postoperative recovery of 6MWD. PMID- 11381502 TI - Comparison of the clinical profile and outcome for squamous cell carcinoma and adenocarcinoma of the distal esophagus and cardia in India. AB - This retrospective review aimed to assess the clinical profile and outcome of squamous cell carcinoma as compared with adenocarcinoma of the lower third of esophagus and cardia following a transhiatal esophagectomy. A total of 169 patients were analyzed retrospectively in this series from 1989 to 1994. There were 100 patients with squamous cell carcinoma (SCC) and 69 patients with adenocarcinoma (ADC). All tumors were assessed by an esophagogram, upper gastrointestinal endoscopy, and abdominal ultrasonography. The surgical procedure performed in all cases was a transhiatal esophagectomy (THE). The mean age of the patients with SCC and ADC was comparable (48 +/- 14 vs 54 +/- 12 years). Male/female ratio was 1.0:1.4 in the SCC group while in the ADC group it was 8.8:1.0. The main symptom in both the groups was grade II dysphagia (62% in SCC and 60% in ADC). The mean length of the tumor was 6.6 +/- 4.5 cm in the SCC group and 4.2 +/- 3.3 cm in the ADC group. The resectability rate of the SCC group was significantly higher (76%) than in the ADC group (55%). The 6-month and 1-year survival for the SCC patients was not significantly different from the ADC patients (83.7% and 49.3% vs 85.0% and 54.0%). The 5-year survival achieved in SCC was higher than in the ADC group (11.6% vs 7.2%) but the difference was not statistically significant. Adenocarcinoma arising from the distal esophagus and cardia was more common in males, and also occurred in a higher age group and had a lower resectability rate than squamous cell carcinoma. No case of Barrett's esophagus was encountered. The short- and long-term survival in both tumors were similar. PMID- 11381503 TI - Microsatellite instability of colorectal cancer and adenoma in synchronous multiple colorectal cancer patients with associated extracolonic malignancies. AB - We examined microsatellite instability (MSI) in nonfamilial multiple synchronous colorectal cancer (multiple CC) patients. We divided the patients into two groups, those with and without extracolonic primary malignancies, and compared the frequency of MSI between the two groups. A colectomy was performed in 52 multiple CC patients between 1985 and 1998. Of them, 10 patients had extracolonic malignancies, while the other 42 patients did not. The MSI frequency was higher in the patients with extracolonic malignancies than in those without extracolonic malignancies, although it was not statistically significant (40% vs 19%, P = 0.21). Regarding the lesions, MSI frequency of cancers was higher in the multiple CC with extracolonic malignancies than in those without extracolonic malignancies (33% vs 13%, P = 0.033). From our results, there was statistically no difference in the existence of extracolonic malignancies between the patients with at least one MSI-positive cancer and those patients without any MSI-positive cancers. On the other hand, there was a significant correlation between MSI-positivity and the existence of extracolonic malignancies. PMID- 11381504 TI - Midcalf ultrasonography for the diagnosis of ruptured Baker's cysts. AB - Clinically significant and palpable enlargement of the gastrocnemio semimembranosus bursa is known as a Baker's cyst. Baker's cysts may rupture, resulting in a swollen, painful leg that is clinically indistinguishable from acute deep vein thrombosis. For this reason, ruptured Baker's cysts are sometimes called pseudothrombophlebitis. The purpose of this study was to determine the incidence of ruptured Baker's cysts, and to evaluate the role of ultrasonography in the diagnosis of this condition. The hospital records of 106 patients (43 men and 63 women) who were referred to the vascular surgical department at Saitama Medical Center with unilateral or bilateral swollen legs between June 1997 and June 2000 were reviewed retrospectively. The total number of affected limbs was 125, being 52 right legs and 73 left legs. Deep vein thrombosis was the most common cause of swollen legs, being diagnosed in 44.8%. No specific anatomical derangement was found in 39 limbs (31.2%), and these were defined as idiopathic. Lymphedema was also common, being diagnosed in 16 limbs. Ruptured Baker's cysts were observed in three limbs, with an incidence of 2.4%. In all of these patients, a large hypoechoic space was seen behind the calf muscles and this sonolucent area was easily detected by a conventional scanner, being pathognomonic of a ruptured Baker's cyst. PMID- 11381505 TI - Antimetastatic effect of hepatotropic liposomal adriamycin on human metastatic liver tumors. AB - Adriamycin (ADM) was encapsulated in a galactose-conjugated hepatotropic liposome (hLip-ADM) and its ability to enhance the antitumor effect while reducing toxicity was compared with that of free ADM and a control Lip-ADM (cLip-ADM), in two in vitro animal models. Liver metastases were induced in rats by an intravenous injection of 8 x 10(6) AH-130 rat hepatoma tumor cells. All forms of the ADM completely inhibited liver metastasis when given at a dose of 5 mg/kg on day 14 after tumor implantation, whereas liver metastatic foci were observed in six of ten rats in the control group. The reduction in ADM toxicity by liposomalization was remarkable, as significant body weight loss was observed only in the free ADM group, in which four of ten rats died. Additional experiments utilized a human colon cancer xenograft (TK-4) to induce the growth of the liver metastases in mice by orthotopic implantation. The hLip-ADM completely inhibited liver metastasis in rats (0/11), whereas liver metastases developed in 10 of 12 mice in the control group and in 5 of 12 mice given cLip ADM. Interestingly, liposomal ADM did not have a significant inhibitory effect on transplanted tumor growth assessed 6 weeks after transplantation. These findings indicate that hLip-ADM may be an effective strategy for inhibiting liver metastases from human colon cancer. PMID- 11381506 TI - Replacement of the canine inferior vena cava with a seeded graft. AB - The patency and microscopic findings of gelatin-coated Dacron grafts seeded with endothelial cells and implanted into the inferior vena cava of dogs was evaluated. A total of 25 mongrel dogs were divided into four experimental groups according to whether or not an antiplatelet agent was administered and a seeded or nonseeded graft was implanted. In the groups not given antiplatelet therapy that were implanted with a nonseeded graft, occlusion of the graft occurred soon after surgery. In contrast, in the majority of animals implanted with a seeded graft and given antiplatelet therapy, graft patency was evident for up to 4 weeks postoperatively. In the latter animals, scanning electron microscopy 4 weeks after surgery showed the formation of confluent endothelial-like cells within 5 mm from the anastomosis, as well as insular endothelial-like cells in the central part of the graft. The simple centrifugal seeding method enables endothelial-like cells to adhere to grafts, suggesting that it may be useful in venous replacement. PMID- 11381507 TI - Thymic carcinoid in a patient with multiple endocrine neoplasia type 1: report of a case. AB - We report herein the rare case of a 33-year-old man found to have a multiple endocrine neoplasia type 1 (MEN1)-associated carcinoid tumor in the thymus. A chest roentgenogram demonstrated an asymptomatic anterior mediastinal mass, 7 cm in diameter, and ultrasound-guided percutaneous Tru-Cut biopsy revealed a carcinoid tumor of the thymus. An extended thymectomy was performed through a median sternotomy and pathological examination confirmed the diagnosis of a thymic carcinoid tumor, which was mainly encapsulated with locally invasive growth into the pleura. Despite the absence of a family history of MEN1, he was treated for two pancreatic islet cell tumors, hyperparathyroidism, an adrenal tumor, and a retroperitoneal lipoma. MEN1 mutations were detected both in blood samples and pancreatic tumor tissues. He is now well without any evidence of tumor recurrence 27 months after the operation for the thymic carcinoid. MEN1 mutations were screened by direct nucleotide sequencing of all protein-coding regions of exons 2-10 of the MEN1 gene. Heterozygous germline mutation was detected in the blood sample analyses. Moreover, fresh-frozen pancreatic tumor tissues showed a loss of heterozygosity in the MEN1 region. PMID- 11381508 TI - Giant mammary hamartoma diagnosed by stereomicroscopic analysis of the mammary glandular tree in an adolescent girl: report of a case. AB - This report describes a rare case of a giant hamartoma that developed in the right breast of a 17-year-old girl. No abnormalities were found by endocrinological studies and a well-circumscribed tumor, approximately 20 cm in diameter, was easily enucleated without bleeding during surgery, following which the bilateral breasts became nearly symmetrical. Histologic features revealed predominant fibrous stroma and scattered normal or occasionally dysplastic mammary glands without neoplastic properties. No distorted lobular structures indicating fibroadenoma characteristics were observed. Subgross and stereomicroscopic analysis of serial 2-mm-thick sections revealed mature normal lobules and predominant fibrous interstitial components. Therefore, the tumor was diagnosed as a giant hamartoma of the breast, according to the histologically non neoplastic features and the delimited macroscopic appearance. This is a rare mammary gland disease characterized by the fact that although each of the histological components seemed to be normal, their constitution was abnormal. It appears that not only histological features but also clinical details are indispensable for the diagnosis of this disease. PMID- 11381509 TI - Pulmonary blastoma: report of two cases. AB - Pulmonary blastomas are a group of rare malignant neoplasms subdivided into three categories: classic biphasic pulmonary blastoma (CBPB), well-differentiated fetal adenocarcinoma (WDFA), and pleuropulmonary blastoma (PPB). We report herein the cases of two men with CBPB. Both were heavy smokers and presented with a history of hemoptysis. Physical examination revealed slightly significant findings, chest radiographs showed a large pulmonary mass, confirmed by computed tomography, and bronchoscopic biopsies were not diagnostic. A left and right inferior lobectomy was performed and a diagnosis of CPBP was confirmed by histological examination. In the first patient, local recurrence with multiple bilateral lung metastases was found 6 months later and despite chemotherapy, he died of respiratory failure 1 year after his operation. In the second patient, a subcutaneous metastasis was found in the right subscapular region 2 months later, and a cerebral metastasis in the right posterior parietal lobe 4 months later. Partial remission was achieved by cerebral irradiation, but 6 months later the patient died of cardiac failure while in a coma. We conclude that more aggressive and multidisciplinary treatment should be adopted for CBPB, and because of its low incidence, it is important to unify individual experiences in a central registry to gather as much information as possible regarding the biological and clinical features of this unusual disease. PMID- 11381510 TI - Primary lung cancer occurring concomitantly with the cicatrized and calcified ova of a parasite: report of a case. AB - We report herein a rare case of primary lung cancer that occurred concomitantly with the calcified ova of a parasite. A 58-year-old man was referred to our department after a pulmonary abnormal shadow had been seen on a chest X-ray done at mass screening. A transbronchial lung biopsy (TBLB) revealed the calcified ova of a parasite. Because the possibility of concomitant lung cancer could not be ruled out, a lung biopsy was taken via video-assisted thoracic surgery (VATS). The pathological diagnosis was squamous cell carcinoma, and a left upper lobectomy was serially performed through a posterolateral thoracotomy. The patient recovered uneventfully and has remained in good health without any sign of recurrence for over 9 months. Following this case report, we review three other cases of this unusual disease combination documented in the literature. PMID- 11381511 TI - Congenital bronchoesophageal fistula with Crohn's disease in an adult: report of a case. AB - A 65-year-old man was diagnosed to have Crohn's disease in 1989. In 1991, frequent bronchitis developed, and bronchoesophageal fistula was diagnosed by esophagography. On esophagography and aortography, the disease was diagnosed to be Type IV based on Braimbridge's classification accompanied by pulmonary sequestration. A thoracoscope-assisted fistelectomy was performed. This paper reports the findings of a fistelectomy assisted by thoracoscopy for the treatment of a Type IV bronchoesophageal fistula according to Braimbridge's classification accompanied by Crohn's disease. PMID- 11381512 TI - A collision tumor composed of adenocarcinoma and malignant lymphoma in the remnant stomach after pancreatoduodenectomy: report of a case. AB - The occurrence of a collision tumor in the stomach, consisting of adenocarcinoma and malignant lymphoma, is extremely rare. We report herein the case of a patient who had undergone a pancreatoduodenectomy for bile duct cancer 5 year earlier, in whom an ulcerating tumor of the remnant stomach developed and grew rapidly within 5 months. Surgical exploration revealed a tumor in the remnant stomach, multiple liver metastases, and multiple lymph node metastases. Total resection of the remnant stomach was performed, and pathological examination revealed a collision tumor consisting of adenocarcinoma and malignant lymphoma. The patient died of liver metastases and lymph node metastases 7 months after his second operation. The coexistence of both adenocarcinoma and malignant lymphoma of the remnant stomach and the etiology of this unusual combination, never previously reported, is discussed. PMID- 11381513 TI - Incarcerated paraesophageal hernia associated with perforation of the fundus of the stomach: report of a case. AB - We report herein a case of incarcerated paraesophageal hernia associated with perforation of the fundus of the stomach. A 71-year-old woman was transferred to our hospital after a diagnosis of gastrointestinal tract perforation had been made at a local hospital. Her history included an esophageal hiatal hernia. A laparotomy was performed which revealed that the antrum of stomach and the duodenal bulb had prolapsed into the esophageal hiatus and become incarcerated. This prolapse had caused stenosis in the corpus of the stomach, resulting in distension of the oral side of the stomach and thinning of the wall. A perforation, 15 mm long, was recognized in the major curvature of the fundus. The patient suffered respiratory failure postoperatively, necessitating respiratory support for 1 week. She was discharged on postoperative day 40. This case report serves to demonstrate that because of the very serious complications that may result from an untreated paraesophageal hernia, elective repair should be performed wherever possible even in asymptomatic patients. PMID- 11381514 TI - Migration of steel-wire coils into the stomach after transcatheter arterial embolization for a bleeding splenic artery pseudoaneurysm: report of a case. AB - Transcatheter arterial embolization (TAE) represents the primary, and often definitive, mode of therapy for bleeding splanchnic artery pseudoaneurysms (PSA). Nevertheless, a number of complications associated with this procedure have been described. We report herein the case of a 59-year-old man with chronic pancreatitis who was referred to us with hematemesis and hemorrhagic shock. Computed tomography revealed a splenic artery PSA bleeding into a pancreatic pseudocyst, and TAE was performed using steel-wire coils, placed inside the aneurysmal cavity, which resulted in the immediate cessation of bleeding. However, several weeks later some of the coils were found to have dislodged through a gastropseudocystic fistula. Furthermore, an early gastric cancer was incidentally found proximal to the fistula. We finally performed open surgery to treat both disorders; primarily for the gastric cancer, but also for the pseudocyst and fistula, with the intermittent discharge of the steel-wire coils. To our knowledge, migration into the stomach of steel-wire coils after TAE has not been described before. It is generally believed that the embolization procedure should occlude normal portions of the artery both distal and proximal to the PSA with embolization materials. By occluding the PSA in this way, the subsequent migration of steel-wire coils into the pseudocyst and stomach might have been prevented in our patient. PMID- 11381515 TI - Splenic and pulmonary metastases from renal cell carcinoma: report of a case. AB - We report herein the case of a patient in whom pulmonary and splenic metastases from renal cell carcinoma (RCC) were successfully treated by surgical excision. A 69-year-old man who underwent left nephrectomy for RCC 17 months before was suspected to have a pulmonary metastasis based on computed tomography (CT) findings. Partial resection of the left lower lobe was performed with thoracoscopic assistance. However, 4 months later, a splenic tumor, 6 cm in diameter, was detected by CT and ultrasonography, and a splenectomy was performed. Histologically, both resected specimens were diagnosed as metastasis from RCC. A second pulmonary metastasis of the left upper lobe was resected 4 years 8 months later. The patient was in good health when last seen 11 months after his last operation. Malignant neoplasms rarely metastasize to the spleen and most cases are found at autopsy, or feature multiple distant metastases. Only four other cases of splenic metastases from RCC have been reported. The prognosis associated with splenic metastasis is favorable when only a solitary lesion exists. PMID- 11381516 TI - Postcoital vaginal laceration in a patient presenting with signs of small bowel perforation: report of a case. AB - Postcoital vaginal rupture or tear is a well-known entity to the gynecologist, albeit unusual; however, such cases are rarely encountered by the general surgeon. The index case is reported to highlight the rare situation wherein a middle-aged woman underwent laparotomy for a suspected small bowel perforation, which revealed a vaginal tear as the cause of pneumoperitoneum. This case emphasizes the importance of taking a gynecological history and performing a gynecological examination when the clinical diagnosis is uncertain. PMID- 11381517 TI - High-output heart failure caused by a huge renal arteriovenous fistula after nephrectomy: report of a case. AB - Postnephrectomy renal arteriovenous fistula (AVF) with an aneurysmal lesion is a rare clinical entity that may cause high-output heart failure. In this report, we describe the case of a 68-year-old man who had undergone nephrectomy for renal tuberculosis 43 years previously, in whom an acquired large renal AVF presenting as an aneurysm caused congestive cardiac failure. We also discuss the hemodynamic, hormonogenic (human arterial natriuretic polypeptide; hANP), and radiographic findings before and after surgery for the AVF. The AVF with an aneurysmal lesion was clearly visualized by three-dimensional-computerized tomographic (CT) scanning, and proximal ligation of the renal artery was followed by an uneventful recovery. This procedure can produce good results when a fistula is too large to allow safe embolization and when excision would be hazardous due to inflammation surrounding the fistula. PMID- 11381518 TI - Delayed presentation of superficial femoral artery injury: report of a case. AB - We describe herein a patient who developed serious complications following a penetrating injury to the lower limb. There was minimal evidence of vascular injury on the initial presentation at the hospital; in particular the ankle systolic pressure was normal. Fourteen days following the initial injury, he was found to have a pseudoaneurysm of the superficial femoral artery associated with the arteriovenous fistula in his left thigh. The findings of this case suggest that a high index of suspicion and a careful clinical review is essential if vascular injuries and their complications are not to be missed. PMID- 11381519 TI - Simple fast graft-to-vessel connection preventing blood leakage, stricture, and aneurysm formation (Ratner-connection). AB - A new simple method of graft-to-vessel anastomosis using a stent technique that prevents blood leakage, stricture, and aneurysm formation is proposed herein. The advantages and problems related to its usage are also discussed. PMID- 11381520 TI - Structural transitions in neutral and charged proteins in vacuo. AB - In vacuo proteins provide a simple laboratory to explore the roles of sequence, temperature, charge state, and initial configuration in protein folding. Moreover, by the very absence of solvent, the study of anhydrous proteins in vacuo will also help us to understand specific environmental effects. From the experimental viewpoint, these systems are now beginning to be characterized at low resolution. Molecular dynamics (MD) simulations, in combination with tools for protein shape analysis, can complement experiments and provide further insights on the folding-unfolding transitions of these proteins. We review some aspects of this issue by using the results from a detailed MD study of hen egg white lysozyme. For lysozyme ions, unfolding can be triggered by Coulombic repulsion. In neutral lysozyme, unfolding can be induced by centrifugal forces and also by weakening the monomer-monomer interaction. In both cases, the resulting unfolded transients can be used as initial configurations for relaxation dynamics. All trajectories are analyzed in terms of global molecular shape features of the backbone, including its anisometry and chain entanglement complexity. This strategy allows us to quantify separately the degree of polymer collapse and the evolution of large-scale folding features. Using these last two notions, we discuss some basic questions regarding the nature of the accessible paths associated with unfolding from, and refolding into, compact conformers. PMID- 11381521 TI - Capture and identification of folding intermediates of cystinyl proteins by cyanylation and mass spectrometry. AB - Trapping folding intermediates of cystinyl proteins by covalent modification of free sulfhydryl groups provides the opportunity for isolation, purification, and structural elucidation of individual species. The disulfide structure of the intermediates, coupled with their temporal abundance, provides a 'snapshot' of the pathway experienced by the refolding protein in a particular medium. Here, intermediates of cystinyl proteins containing free cysteines are trapped by cyanylation through reaction with an acidic (pH 3.0) solution of 1-cyano-4 dimethylamino-pyridinium (CDAP) tetrafluoroborate. The cyanylated species are separated by reversed-phase high-performance liquid chromatography, where the resulting chromatogram gives a visual indication of the distribution of intermediates at a designated time after commencing the refolding process. The disulfide structure of an intermediate can be determined by cleaving its cyanylated derivative and by mass mapping of the resulting fragments to the sequence of the original protein. Cleavage of a cyanylated species represented by any given peak in the chromatogram is achieved by treatment of that fraction with 1M NH4OH at room temperature for 1 h; the resulting fragments are analyzed by matrix-assisted laser desorption ionization (MALDI) or electrospray mass spectrometry. Examples will be presented from in vitro refolding experiments with human epidermal growth factor (hEGF), for which more than 10 folding intermediates were isolated and identified at different time points, and a mutant of insulin-like growth factor-I, for which three intermediates were isolated and identified. PMID- 11381522 TI - Solid state NMR measurements of conformation and conformational distributions in the membrane-bound HIV-1 fusion peptide. AB - The solid state NMR lineshape of a protein backbone carbonyl nucleus is a general diagnostic of the local conformational distribution in the vicinity of that nucleus. In addition, measurements of carbonyl chemical shifts and 2D exchange spectra provide information about the most probable conformation in the distribution. These types of solid state NMR methodologies have been applied to structural studies of the membrane-bound HIV-1 fusion peptide. This peptide is derived from a domain of the HIV-1 gp41 envelope protein, which is critical for viral-host cell-membrane fusion. Even in the absence of the rest of the envelope protein, the fusion peptide will fuse liposomes or erythrocytes. The solid state NMR measurements demonstrate that the center of the membrane-bound HIV-1 fusion peptide is structured, while the C-terminus is highly disordered. The structural distribution at the peptide center is lipid-dependent, with the greatest degree of structural homogeneity in a lipid environment whose composition reflects that of the target T cells. When bound to the lipid mixture, the peptide center is predominately beta sheet. The beta-sheet structure may be diagnostic of peptide oligomerization, which is thought to be a requirement for membrane fusion activity. Although the peptide partially disrupts bilayer orientational ordering in stacked glass-plate samples, 2H NMR demonstrates that the bilayers remain intact in the presence of the fusion peptide and are not micellized. The retention of the bilayer phase may relate to the biological requirement that the virus should fuse with, but not destroy, the target host cell membrane. PMID- 11381523 TI - Steered molecular dynamics investigations of protein function. AB - Molecular recognition and mechanical properties of proteins govern molecular processes in the cell that can cause disease and can be targeted for drug design. Single molecule measurement techniques have greatly advanced knowledge but cannot resolve enough detail to be interpreted in terms of protein structure. We seek to complement the observations through so-called Steered Molecular Dynamics (SMD) simulations that link directly to experiments and provide atomic-level descriptions of the underlying events. Such a research program has been initiated in our group and has involved, for example, studies of elastic properties of immunoglobulin and fibronectin domains as well as the binding of biotin and avidin. In this article we explain the SMD method and suggest how it can be applied to the function of three systems that are the focus of modern molecular biology research: force transduction by the muscle protein titin and extracellular matrix protein fibronectin, recognition of antibody-antigene pairs, and ion selective conductivity of the K+ channel. PMID- 11381524 TI - Continuum solvent molecular dynamics study of flexibility in interleukin-8. AB - Generalized Born continuum solvent methods have been shown to provide a reasonable description of the equilibrium thermodynamics of aqueous solvation in a variety of applications to peptides, proteins, and nucleic acids. Here we study the performance of these methods in molecular dynamics simulations of interleukin 8, comparing nanosecond-length explicit solvent simulations with those using the generalized Born model. In general, the simulations show similar results, although movement away from the initial NMR-determined structure and average fluctuations about the mean are slightly higher for the continuum solvent results. In both simulations, the two helices that are packed on top of the core sheet move closer together, resulting in a structure that more closely resembles the X-ray structure. Principal-component (quasiharmonic) analysis is used to analyze the motions of these helices in both of the simulations and in the NMR ensemble of structures. Prospects for making more general use of continuum solvent models in protein dynamics simulations are discussed. PMID- 11381525 TI - State of the art in studying protein folding and protein structure prediction using molecular dynamics methods. AB - This study presents an overview of the state of the art in using molecular dynamics methods to simulate protein folding and in the end game of protein structure prediction. In principle, these methods should allow the highest level of detail possible and the highest accuracy, but they are limited by both the accuracy of the force field used in the simulation and the sampling possible in the available computer time. We describe current capabilities in running the simulations longer and more efficiently. PMID- 11381526 TI - Evolution of functionality in lattice proteins. AB - We study the evolution of protein functionality using a two-dimensional lattice model. The characteristics particular to evolution, such as population dynamics and early evolutionary trajectories, have a large effect on the distribution of observed structures. Only subtle differences are observed between the distribution of structures evolved for function and those evolved for their ability to form compact structures. PMID- 11381527 TI - The designability of protein structures. AB - It has been noted that natural proteins adapt only a limited number of folds. Several researchers have investigated why and how nature has selected this small number of folds. Using simple models of protein folding, we demonstrate systematically that there is a "designability principle" behind nature's selection of protein folds. The designability of a structure (fold) is measured by the number of sequences that can design the structure--that is, sequences that possess the structure as their unique ground state. Structures differ drastically in terms of their designability. A small number of highly designable structures emerge with a number of associated sequences much larger than the average. These highly designable structures possess proteinlike secondary structures, motifs, and even tertiary symmetries. In addition, they are thermodynamically more stable and fold faster than other structures. These results suggest that protein structures are selected in nature because they are readily designed and stable against mutations, and that such a selection simultaneously leads to thermodynamic stability. PMID- 11381528 TI - Comparing protein structures: a Gaussian-based approach to the three-dimensional structural similarity of proteins. AB - This study describes a new method for comparing three-dimensional protein structures based on an optimal alignment of their steric fields. The method is based upon the use of spherical Gaussian functions located on individual atoms. This representation generates a flexible description of the underlying fold geometry of proteins that can be adjusted by changing the 'width' of the Gaussians. Reducing the width sharpens the representation and leads to a more 'atomlike' description; increasing the width creates a fuzzier representation that preserves the general shape features of the chain fold but with a consequent loss in atomic resolution. The width used in this study is based upon the features of individual atoms and provides a representation that is quite robust with respect to the variety of geometric features typically encountered in the alignment process. In addition, a post-alignment analysis is performed that generates sequence alignments from the corresponding structure alignments. An example, based on five mammalian and fungal matrix metalloproteinase crystal structures (human fibroblast collagenase, neutrophil collagenase, stromelysin, astacin, and adamalysin), illustrates a number of features of the Gaussian-based approach. PMID- 11381530 TI - NMR spin relaxation methods for characterization of disorder and folding in proteins. AB - The flexibility and dynamics of proteins directly influence the processes of protein folding, recognition, and function. NMR spin relaxation methods are used to assess the dynamics and mobility of proteins, for fast ps and ns motions as well as slower microsecond and ms events. The degree of protein flexibility and disorder as well as the changes in protein flexibility can be assessed by NMR spin relaxation methods at individual residues within the protein. In addition to probing protein dynamics, the changes in the NMR-derived order parameters can be used to estimate the entropic contributions of order-disorder transitions. Furthermore, kinetic processes in the ms time regime may be directly investigated to extract the rates of conformational interconversion, ligand binding, and protein folding processes. We show how a variety of dynamical information can be obtained from NMR relaxation measurements. We present examples that illustrate the use of NMR spin relaxation analysis for investigation of folding and disorder in proteins. PMID- 11381529 TI - Intrinsically disordered protein. AB - Proteins can exist in a trinity of structures: the ordered state, the molten globule, and the random coil. The five following examples suggest that native protein structure can correspond to any of the three states (not just the ordered state) and that protein function can arise from any of the three states and their transitions. (1) In a process that likely mimics infection, fd phage converts from the ordered into the disordered molten globular state. (2) Nucleosome hyperacetylation is crucial to DNA replication and transcription; this chemical modification greatly increases the net negative charge of the nucleosome core particle. We propose that the increased charge imbalance promotes its conversion to a much less rigid form. (3) Clusterin contains an ordered domain and also a native molten globular region. The molten globular domain likely functions as a proteinaceous detergent for cell remodeling and removal of apoptotic debris. (4) In a critical signaling event, a helix in calcineurin becomes bound and surrounded by calmodulin, thereby turning on calcineurin's serine/threonine phosphatase activity. Locating the calcineurin helix within a region of disorder is essential for enabling calmodulin to surround its target upon binding. (5) Calsequestrin regulates calcium levels in the sarcoplasmic reticulum by binding approximately 50 ions/molecule. Disordered polyanion tails at the carboxy terminus bind many of these calcium ions, perhaps without adopting a unique structure. In addition to these examples, we will discuss 16 more proteins with native disorder. These disordered regions include molecular recognition domains, protein folding inhibitors, flexible linkers, entropic springs, entropic clocks, and entropic bristles. Motivated by such examples of intrinsic disorder, we are studying the relationships between amino acid sequence and order/disorder, and from this information we are predicting intrinsic order/disorder from amino acid sequence. The sequence-structure relationships indicate that disorder is an encoded property, and the predictions strongly suggest that proteins in nature are much richer in intrinsic disorder than are those in the Protein Data Bank. Recent predictions on 29 genomes indicate that proteins from eucaryotes apparently have more intrinsic disorder than those from either bacteria or archaea, with typically > 30% of eucaryotic proteins having disordered regions of length > or = 50 consecutive residues. PMID- 11381531 TI - Protein flexibility and dynamics using constraint theory. AB - A new approach is presented for determining the rigid regions in proteins and the flexible joints between them. The short-range forces in proteins are modeled as constraints and we use a recently developed formalism from graph theory to analyze flexibility in the bond network. Forces included in the analysis are the covalent bond-stretching and bond-bending forces, salt bridges, and hydrogen bonds. We use a local function to associate an energy with individual hydrogen bonds, which then can be included or excluded depending on the bond strength. Colored maps of the rigid and flexible regions provide a direct visualization of where the motion of the protein can take place, consistent with these distance constraints. We also define a flexibility index that quantifies the local density of flexible or floppy modes, in terms of the dihedral angles that remain free to rotate in each flexible region. A negative flexibility index provides a measure of the density of redundant bonds in rigid regions. A new application of this approach is to simulate the maximal range of possible motions of the flexible regions by introducing Monte Carlo changes in the free dihedral angles, subject to the distance constraints. This is done using a method that maintains closure of the rings formed by covalent and hydrogen bonds in the flexible parts of the protein, and van der Waals overlaps between atoms are avoided. We use the locus of the possible motions of HIV protease as an example: movies of its motion can be seen at http://www.pa.msu.edu/~lei. PMID- 11381532 TI - Structure and dynamics of 6-hydroxymethyl-7,8-dihydropterin pyrophosphokinase. AB - Folates are essential for life. Unlike mammals, most microorganisms must synthesize folates de novo. 6-Hydroxymethyl-7,8-dihydropterin pyrophosphokinase (HPPK) catalyzes pyrophosphoryl transfer from ATP to 6-hydroxymethyl-7,8 dihydropterin (HP), the first reaction in folate pathway, and therefore, is an ideal target for developing novel antimicrobial agents. Because of its small size and high thermal stability, E. coli HPPK is also an excellent model enzyme for studying the mechanisms of enzymatic pyrophosphoryl transfer. We have determined the crystal structures of HPPK in the unligated form and in complex with HP, two Mg2+ ions, and AMPCPP (an ATP analog that inhibits the enzymatic reaction). Comparison of the two crystal structures reveals dramatic conformational changes of three flexible loops and many side chains and possible roles of the active site residues. PMID- 11381533 TI - Sampling activated mechanisms in proteins with the activation-relaxation technique. AB - The activated dynamics of proteins occur on time scales of milliseconds and longer. Standard all-atom molecular dynamics simulations are limited to much shorter times, of the order of tens of nanoseconds. Therefore, many activated mechanisms that are crucial for long-time dynamics will not be observed in such molecular dynamics simulation; different methods are required. Here, we describe in detail the activation-relaxation technique (ART) that generates directly activated mechanisms. The method is defined in the configurational energy landscape and defines moves in a two step fashion: (a) a configuration is first brought from a local minimum to a nearby first-order saddle point (the activation); and (b) the configuration is relaxed to a new metastable state (the relaxation). The method has already been applied to a wide range of problems in condensed matter, including metallic glasses, amorphous semiconductors and silica glass. We review the algorithm in detail, discuss some previously published results and present simulations of activated mechanisms for a two-helix bundle protein using an all-atom energy function. PMID- 11381534 TI - Constructing smooth potential functions for protein folding. AB - A protein folding potential function ideally has several properties: it favors the native conformations for a number of protein sequences over a variety of nonnative folds; it can guide the search over conformations for the native state; it reflects changes in stability of the native fold due to changes in sequence; and it is relatively insensitive to small changes in conformation. While these are not mutually incompatible goals, attaining one property does not ensure that the others are satisfied. Examples are given of simple potentials having one property but lacking others. A new functional form of a folding potential is described where interactions between all nonhydrogen atoms are used to estimate interresidue interactions and implicit solvation. Its parameters can be adjusted to satisfy the above properties at least for barnase and a few other proteins. PMID- 11381535 TI - Experimental approaches to protein folding based on the concept of a slow hydrogen exchange core. AB - In a review of protein hydrogen exchange, we concluded that the slow exchange core is the folding core. By this we mean that the elements of secondary structure carrying the slowest exchanging backbone amides will tend to be the elements of secondary structure to fold first, that partially folded proteins will tend to be most organized in the core, and that peptides made to mimic the slow exchange core will tend to show nativelike structure. These generalizations have led us to ask several experimental questions that will be examined here: (1) In partially folded and unfolded proteins, how do the dynamics and structure of core regions differ from noncore regions? (2) Can we make protein 'core modules' as peptides corresponding to the slow exchange core? Can core modules be covalently linked to make a native state in which one conformation is significantly more stable than all other accessible conformations? (3) In a mutant perturbed outside the core, what are the effects on hydrogen exchange and folding? PMID- 11381537 TI - Cationic membranes for phenols production. AB - Ionic Substitution by Electrodialysis is a good alternative to the industrial conversion of sodium salts of phenols into undissociated phenols, conventionally performed through acidification by strong acids. The acidification through electromembranes combines the possibility to achieve high conversion values with the advantage of keeping the process stream separated from the acid stream. The process is performed through cationic membranes; conversion can be obtained also in the absence of electric current, even if slightly better performances can be achieved by applying an electric field. Process feasibility as well as membrane resistance is tested, for the case of model solutions reproducing the main features of the real process streams. Maximum conversion and process rate are greatly affected by the ratio between protons in the acid solution and sodium ions in the process solution. Phenol losses into the acid stream and chloride contamination of the process stream can be reduced by working with diluted process and acid streams. PMID- 11381536 TI - Preparation and characterisation of membranes with entrapped TiO2 and preliminary photocatalytic tests. AB - The possibility of entrapping polycrystalline TiO2 in a polymeric support, in order to couple the two unit operations, i.e. ultrafiltration and photodegradation of organic pollutants in aqueous solutions, was investigated. To this aim, polymeric membranes for ultrafiltration with entrapped TiO2 were prepared, characterised and tested. The polymeric support chosen was commercial polysulfone (PSf). The membrane preparation was carried out with the technique referred to as phase inversion. A three-component system, with a polymer, a solvent and a non-solvent was used. The best operative conditions were determined in order to obtain the desired membrane morphology. Permeability measurements and photostability tests were also carried out by using a system under pressure. Finally, a preliminary investigation was performed in order to evaluate the photocatalytic activity of the membranes with entrapped TiO2 for the oxidation of 4-nitrophenol as a model molecule in aqueous solutions. PMID- 11381538 TI - Innovative methods for removing phosphorus from wastewaters. AB - Wastewater containing a mixture of hypophosphites and phosphites was treated in order to assess the possibility of attaining high levels of phosphorus oxidation to phosphate. An initial series of tests based on chemical oxidation by means of Fenton's reagent demonstrated the feasibility of this process for solutions containing 250-750 mg/l of phosphorus. Removal yields of up to 98% were attained at pH 3.5 by using variable ratios of [H2O2]/[Fe2+]. The second series involved the electrolytic generation of Fenton's reagent at pH 2-2.5 through cathodic oxygen reduction. This demonstrated the possibility of obtaining quantitative yields for solutions containing 80-160 mg/l P with a constant [H2O2]/[Fe2+] ratio of 3.94. Economic comparison of the two processes showed that the operative cost of the electrochemical treatment is comparable to that of the chemical process. PMID- 11381539 TI - Organic halide electroreduction on silver. AB - Silver, whose extraordinary electrocatalytical properties for organic halide reduction have been recently evidenced, has been used as cathode material for systematic preparative electrolyses in membrane-divided cells. To better elucidate the substrate role on the remarkable positive shift of reduction potentials, and on the "cage effect" i.e. the promotion of intermolecular reaction on adsorbed intermediates, three halide substrate patterns are here compared in terms of both voltammetric characterization and preparative electroreduction products: aliphatic halides (adamantanes), aromatic halides (phenols) and anomeric glycosyl halides. The preparative electroreductions result mainly in dimerization in the case of glycosyl halides, in H-->Br substitution in the case of bromophenols, in dimerization + substitution in the case of haloadamantanes. The product analysis, both at the end of the reaction and at intermediate times, allows discussing the reaction pathways in terms of intermediate stability and of active surface accessibility. The possibility of complete dehalogenation on a wider substrate variety with remarkably lower energy consumption and almost quantitative current yields makes the process potentially very interesting for environmental purposes. PMID- 11381540 TI - Electrochemical oxidation of para-chlorophenol on Ti/SnO2-PbO2 electrodes: introduction of a parameter for the estimation of their efficiency. AB - To oxidate otherwise non-biodegradable organic pollutants as chlorophenol has recently been proposed the promising electro-combustion technique. We considered two different types of electrodic material: boron doped diamond (BDD) and lead dioxide electro-deposited on a Ti/SnO2 substrate and we investigated differences between them in terms of organics oxidation efficiency. In the batch system we utilised for the combustion, the experimental parameters we investigated, initial concentration of the organic compound, temperature and current density furnished to the system--showed us that a parameter like the Instantaneous Current Efficiency (ICE) is affected by mass-transfer limitation at the end of the tests. In this paper we therefore introduce a new parameter not influenced by these problems that can be very effective for the description of electrode efficiency during the conduction of experiences. PMID- 11381541 TI - Electrochemical treatment of olive oil mill wastewater. AB - The possibility of oxidizing at a PbO2 anode the phenols and polyphenols, present in the olive oil mill wastewater, has been studied as a pretreatment for the submission of such wastewater to the traditional biological treatments. The results obtained operating at current densities ranging 500 to 2000 A/m2 show that it is possible to reduce the concentration of the phenolic components, which interfere with the biological treatments, down to low values without decreasing too much the total organic content of the wastewater. PMID- 11381542 TI - The removal of low level inorganics via electrogenerated hydrogen peroxide in the presence of catalytic amounts of Fe2+. AB - Low level phosphites and hypophosphites were completely converted into phosphates, via hydrogen peroxide generated by cathodic reduction of oxygen in acidic aqueous medium at a reticulated vitreous carbon electrode, in the presence of little amounts of Fe2+. The contemporary regeneration of Fe2+ by cathodic reduction of Fe3+, produced by the well known Fenton reaction, furnishes an excellent way to continuously produce little amounts of the Fenton reactive and, as a consequence, of the powerful oxidant hydroxyl radical HO.. The best conditions for the complete removal of phosphorous as phosphites and hypophosphites are reported. PMID- 11381544 TI - Supported liquid membrane stability in chiral resolution by chemically and physically modified membranes. AB - In the present work some stability studies on Supported Liquid Membranes (SLMs) to be used for chiral separations were realized. In particular, primary aim was to determine how a modification of the support surface influences the SLM stability. First, the procedure for support modification was optimised, making a screening of various compounds (sulphuric acid, nitric acid, chromic acid, sodium dodecyl sulphate (SDS), glycerol, oleic alcohol, propylene glycol (PPG), bovine serum albumin (BSA)) and testing their performance by means of contact angle measurements. Next, a second screening was realized by permeation tests in a stirred cell. Finally, to compare the stability of modified with unmodified support in a process of interest for chemical and/or biochemical industries, some permeation tests for resolution of DNB-DL-Leucine were realized in a re circulation system. Results showed a better surface hydrophilization of chemically modified support and better stability of the sulphonated support. However, in operating conditions a little high stability of the unmodified support was obtained. PMID- 11381543 TI - Industrial solution contaminated by polyacrylates: their elimination by electrochemical combustion. AB - The electrochemical combustion of polyacrylates was studied through both direct and indirect oxidation. The obtained results indicate the non elimination of the polyacrylates with the direct oxidation, while the indirect oxidation with NaCl completely eliminates these organic compounds. In the last case the effects of different initial concentrations of NaCl, anode materials and current densities was studied. PMID- 11381545 TI - Photocatalytic membrane processes for degradation of various types of organic pollutants and micropollutants in water. AB - Degradation tests in a photocatalytic membrane system have been carried out using TiO2 (Degussa P25) as catalyst and humic acid, organic dyes, 4-nitrophenol as pollutants. The influence of UV radiation and initial concentration of pollutant on the photodegradation rate were investigated in discontinuous and continuous systems. Experimental results showed that it is possible to obtain an efficient photocatalytic membrane process, but various parameters (e.g. pH) should be optimised to obtain high reaction rate and high membrane rejection of pollutants and their by-products. PMID- 11381546 TI - Sequential electrochemical/biological treatment for the removal of 2,6 dichlorophenol from synthetic wastewater. AB - The paper examines the effect of chloride on the oxidation of 2,6-dichlorophenol (DCP) performed at TiO2/RuO2 DSA anodes, which are specific catalysts for chlorine evolution. The results indicate that chlorine/hypochlorite originating from chloride oxidation in certain favourable conditions reacts with the organic substrate at the diffusion layer near the anode, accelerating the mass transfer of the reactant towards the electrode surface. When the bulk concentration of organic substrate has become very low, the oxidising species can accumulate in the bulk solution where the accomplishment of the oxidation of residual reactant and of its intermediates takes place. Solutions which also contained glucose were electrolysed in order to verify the high level of selectivity of DCP oxidation with respect to a biodegradable substrate: glucose was found to be unchanged up to nearly complete elimination of DCP. The toxicity of the solution was sufficiently reduced to reach values compatible with the subsequent biological treatment. PMID- 11381547 TI - Abatement of waste-water biorefractory organics via electro-oxidative treatment. AB - The electrochemical oxidation of coumaric acid, a biorefractory compound present in several industrial waste waters, has been investigated by use of Pt-Ti anodes and at electrolyte concentration (0.02 N NaCl or Na2SO4) low enough to allow direct dischargeability of the waste water into superficial water basins according to the Italian law (DL 152/11-5-99). Particularly, the role of the electrolyte over the conversion rate has been assessed. The obtained results show that the oxidation process should take place both at the electrode surface and in the bulk of the solution, via electrochemically-generated oxidising species (H2O2, persulfates, Cl2, NaClO). The faster coumaric acid abatement rates were found with chloride based electrolytes, which, however, lead to the formation of non-biodegradable small-molecular-weight chlorinated hydrocarbons. PMID- 11381548 TI - Innovation in sodium erythorbate production: the use of membrane-reactors. AB - Isoascorbic or erythorbic acid is a stereoisomer of ascorbic acid acting as a preservative against oxidation and decoloration, recently approved for food use also in the European Market. Actually erythorbate is produced by means of a complex and long process after bio-fermentation of dextrose. In order to simplify and improve this process a membrane system was conceived able to induce acid 2 ketogluconic diffusion from fermentation broth directly into methanol where it esterificates. Acid 2-ketogluconic methyl ester is the intermediate for erythorbic acid. The principal problem was to perfectly separate water from methanol to avoid saponification instead of esterification. PMID- 11381549 TI - Speciation. PMID- 11381550 TI - Coloured rain dust from Sahara Desert is still radioactive. AB - A coloured rain event originating from the Sahara Desert occurred on April 9, 2000 at Thessaloniki, Northern Greece (40 degrees 38'N, 22 degrees 58'E). The radioactive nuclides that were determined in a coloured rain dust sample were 137Cs of Chernobyl origin, 7Be of cosmogenic origin and 40K of terrestrial origin. Cesium-137 still remained 14 years after the Chernobyl accident, reaching 26.6 Bq kg-1 in the coloured rain dust. PMID- 11381551 TI - 226Ra and 228Ra activities associated with agricultural drainage ponds and wetland ponds in the Kankakee Watershed, Illinois-Indiana, USA. AB - Background radioactivity is elevated in many agricultural drainage ponds and also constructed wetland ponds in the Kankakee watershed. During 1995-1999, gross alpha and -beta activities were measured up to 455 and 1650 mBq L-1, respectively. 226Ra and 228Ra averaged 139 and 192 mBq L-1 in controlled drainage ponds compared to 53 and 58 mBq L-1 for 226Ra and 228Ra, respectively, in native wetland ponds. Analyses of applied ammonium phosphate fertilizers near both native and controlled ponds indicate comparable 226Ra/228Ra and 228Ra/232Th activity ratios with only the surface waters in the controlled ponds. For example, 226Ra/228Ra activity ratios in controlled ponds ranged from 0.791 to 0.91 and group with a local fertilizer batch containing FL phosphate compounds with 226Ra/228Ra activity ratios of 0.831-1.04. Local soils of the Kankakee watershed have 226Ra/228Ra activity ratios of 0.541-0.70. Calculated Ra fluxes of waters, in drainage ditches associated with these controlled ponds, for 226Ra ranged from 0.77 to 9.00 mBq cm-2 d-1 and for 228Ra ranged from 1.22 to 8.43 mBq cm-2 d-1. Ra activity gradients were measured beneath these controlled ponds both in agricultural landscapes and in constructed wetlands, all being associated with drainage ditches. Ra had infiltrated to the local water table but was below regulatory maximum contaminant limits. Still, measurable Ra activity was measured downgradient of even the constructed wetlands in the Kankakee watershed, suggesting that the attenuation of Ra was low. However, no Ra excess was observed in the riparian zone or the Kankakee River downgradient of the native wetland ponds. PMID- 11381552 TI - A 10-year study of the 137Cs distribution in soil and a comparison of Cs soil inventory with precipitation-determined deposition. AB - During a 10-year period, 1988-1998, surface soil samples have been collected at Blentarp in southern Sweden and analysed for 137Cs from the Chernobyl accident and from the nuclear weapons tests. The distance between the sampling plots on the different sampling occasions has been no more than 3 m. The results show that the depth distribution of 137Cs is very similar for each of the sampling occasions, indicating that the caesium migration at this site is very small. The total activity measured in the soil cores is in agreement with the calculated activity of 137Cs deposited at the site after nuclear weapons tests and the Chernobyl accident, based on air activity concentration and the amount of precipitation. The calculated deposition of 137Cs originating from the bomb tests amounts to 1.41 kBq m-2 for the period 1962-1986, which is in agreement with the activity of nuclear weapons fallout measured in the soil samples (1.60 kBq m-2 as a mean value of the first four years of sampling). The calculated activity of 137Cs of Chernobyl origin was 0.79 kBq m-2, which agrees well with the value of 0.79 kBq m-2 measured in the soil samples in 1988. PMID- 11381553 TI - Time-dependence of the radiocaesium contamination of roe deer: measurement and modelling. AB - In spruce forest and peat bog, the migration of 137Cs from soil to plants, fungi, roe deer and consumers has been surveyed. In spruce forest the 137Cs activity concentration in roe deer decreases slowly with time and has superimposed periodic maxima in autumn which are correlated with the mushroom season. The decrease with time can be described by an effective half-life of 3.5 yr caused by a fraction of the 137Cs in the soil becoming unavailable for green grazing plants with time. The additional transfer of 137Cs into roe deer meat during the mushroom season depends on precipitation in July, August and September which also determines the yield of fungi in autumn. Our model confirms the assumption that fungi also have access to a fraction of the 137Cs in the soil which is unavailable for green plants. On peat bog the 137Cs activity concentration in roe deer is higher than in spruce forest and its effective half-life is about 17 yr, due to reversible binding of 137Cs to organic matter in the peat bog. PMID- 11381554 TI - Reinvestigation of airborne 210Pb, 137Cs and 207Bi in Vienna (Austria) after atmospheric nuclear weapons tests. AB - Reconstruction of atmospheric 210Pb and 137Cs activity concentrations occurring in Vienna immediately after the atmospheric nuclear weapons tests shall answer the question, whether artificial 210Pb has been produced by fusion devices using lead materials. If so, 207Bi also can be expected. In 1962, weekly average values for 210Pb were between 0.08 +/- 0.02 and 1.31 +/- 0.30 mBq/m2, 137Cs amounts up to 5.1 +/- 0.6 mBq/m3. 207Bi remains below the detection limit, viz. < 0.5 mBq/m3. Bimonthly averages for the periods 1962-1964 and 1974-1975 show 137Cs activity concentrations with typical springtime maxima occurring together with low 210Pb values and 207Bi < 0.02 mBq/m3. Annual averages of 210Pb are varying between 0.27 and 0.53 mBq/m3, independent of whether atmospheric tests have been performed or not. Comparison of the activity ratios 207Bi/210Pb, 207Bi/137Cs and 210Pb/137Cs with published data, leads to the conclusion that no evidence exists for a significant production of 210Pb by nuclear explosions, neither after the Soviet tests in 1961-1962 nor after the Chinese tests in 1973-1976. PMID- 11381555 TI - Kinetics of the adsorption and desorption of radionuclides of Co, Mn, Cs, Fe, Ag and Cd in freshwater systems: experimental and modelling approaches. AB - The sorption and release kinetics of 54Mn, 58Co, 59Fe, 109Cd, 110mAg and 134Cs by freshwater suspended particles were investigated to better identify the biogeochemical processes involved and to obtain suitable data for improving models describing radionuclide migration in freshwater streams. In order to observe any seasonal variability in the interaction of radionuclides with natural particles, experiments were performed both in winter and in summer during a phytoplanktonic bloom. Two kinetic models are compared in this paper: the "one step reversible" model, based on the hypothesis that the transfer of radionuclides between water and suspended solids is governed by a reversible reaction, and the "two-successive-step reversible" model, which assumes two distinct types of sites or reactions on the solid phase. The "one-step reversible" model is generally unable to describe properly the exchange kinetics; this result shows that at least two processes are generally involved in radionuclide exchange between water and suspended particles. On the contrary, a model involving the existence of two successive reversible reactions properly simultes both the sorption and release kinetics. The determination of the kinetic coefficients allows quantitative assessment of the relative importance and kinetics of the processes. In particular, it has been shown that, except for Cs and Cd, major fractions of the radionuclides are associated at equilibrium with particulate sites involving strong interactions. The kinetics to reach this equilibrium depend on seasonal conditions, especially for Co and Mn: the transfer of Co and Mn to particulate sites involving strong interactions is much slower in winter. The distribution of the radionuclides between water and particulate sites involving weak interactions also shows seasonal variations for Co, Mn, Fe and Ag: the capacity to associate radionuclides is much higher in summer for Co and Mn, while the inverse tendency is observed for Fe and Ag. For Cs and Cd, no significant seasonal differences were observed. PMID- 11381556 TI - Behaviour and transport of radionuclides in soil and vegetation of a sand dune ecosystem. AB - A sand dune ecosystem in the vicinity of the British Nuclear Fuels reprocessing plant at Sellafield, Cumbria, UK was used to examine the spatial, temporal and depth distributions of 134Cs, 137Cs, 238Pu, 239 + 240Pu and 241Am in soil and in two species of vegetation (Festuca rubra, Ammophila arenaria). Core samples showed evidence of the accumulation of radionuclides derived mainly from sea-to land transfer. Accumulated deposits of radioactivity (0-0.1 m) lie within the range: 1.1-3.4 Bq kg-1 (134Cs), 260-440 Bq kg-1 (137Cs), 31-40 Bq kg-1 (238Pu), 150-215 Bq kg-1 (239 + 240Pu) and 190-240 Bq kg-1 (241Am). Soil profiles showed greater activity concentrations in their deeper regions and this is attributed to leaching of radionuclides in percolating drainage water accentuated by the coarse texture, low organic matter and clay mineral content of coastal sands. Radionuclide activity concentrations in F. rubra and A. arenaria were similar, in the ranges 20-70 Bq kg-1 (137Cs), 1-5 Bq kg-1 (238Pu), 10-30 Bq kg-1 (239 + 240Pu) and 10-65 Bq kg-1 (241Am). Clear temporal and spatial variations were observed in both species of vegetation, reflecting the weather conditions antecedent to the sampling period and the influence of sea-to-land transfer. Concentration ratios (vegetation:soil) for activity concentrations in the two species were similar, in the ranges: 0.05-0.14 (137Cs), 0.025-0.097 (238Pu), 0.022-0.057 (239 + 240Pu) and 0.025-0.212 (241Am). PMID- 11381557 TI - Research development at the University of Health Sciences College of Osteopathic Medicine. PMID- 11381558 TI - Letter that questions OCF requires no response. PMID- 11381559 TI - Innovative approach to headaches. PMID- 11381560 TI - More on spirituality. PMID- 11381561 TI - More on spirituality. PMID- 11381562 TI - President Bush's 2002 budget proposal would hamper geriatric education. PMID- 11381563 TI - Segmental definition--Part IV. Updating the differential for somatic and visceral inputs. AB - The first three parts of this series presented a standardized method of motion testing for motor behaviors specific to segmental dysfunction. When applied to clinical research studies, analysis of these palpatory data differentiated viscero-somatic from somato-somatic reflex manifestations in the thoracic region of subjects with renal dysfunction and/or hypertension. This update for the differential adds new observations regarding palpable findings, specifically at the spinal levels where linkage of costal and vertebral motion asymmetries identified the presence of increased visceral inputs. At each level, the examiner can make a simple comparison for accord in response to two sidebending motion tests in the seated position, one introduced through the shoulders and trunk and one through the head and neck. Lack of accord is a distinctive characteristic of the linked spinal/costal site of visceral input. Also, with regard to linkage sites, behavioral responses to motion tests introduced through the lower extremities suggest new applications for manipulative intervention. The new descriptive clinical data about segmental motion behaviors described in this report provide a basis for new questions in neuromusculoskeletal research. PMID- 11381564 TI - Traumatic pseudoaneurysm of the superficial temporal artery: two cases. AB - Pseudoaneurysms of the superficial temporal artery are a rare and potentially critical cause of facial masses. Most pseudoaneurysms form as a result of blunt trauma and present as painless, pulsatile tumors that may be associated with neuropathic findings and enlarged size. Without careful evaluation in the primary care setting, pseudoaneurysms can be easily misdiagnosed and improperly managed. They can, however, be accurately diagnosed through physical examination alone and subsequently treated with surgical ligation. The authors present two cases of traumatic pseudoaneurysms of the superficial temporal artery caused by blunt injury and discuss pertinent diagnosis and treatment options, as well as provide a brief review of the anatomy and histopathology of pseudoaneurysms. PMID- 11381565 TI - Occupational and environmental medicine in a family medicine residency. AB - Well-rounded instruction in occupational medicine as part of family medicine residency training is feasible through a program that balances a longitudinal curriculum of monthly occupational and environmental medicine (OEM) lectures, community-based OEM patient care, and worksite assessment. Such training would help equip family medicine residents to integrate occupational medicine into their practices, which, in light of a shortage of board-certified practitioners in OEM, would help fill community needs. The Intern-Resident Training Committee of Carson City Hospital in rural Michigan established both learner and institutional goals and objectives for such a program of instruction. A learner needs assessment found appreciable interest among the residents for occupational medicine training. In addition, results of a survey of the occupational health community suggested there is inadequate coverage of OEM in medical schools and residencies. Furthermore, a focus group of occupational health managers revealed that clarity of communication and standardization of reporting were paramount concerns. Instruments for standardized OEM history and for OEM case management were developed for use within the residency continuity clinic. The curriculum was implemented with a variety of teaching strategies, including worksite assessment. Practice-based, case-oriented instruction was subsequently phased into the program as residents assumed responsibility for managing cases under the supervision of family medicine preceptors knowledgeable in OEM. An occupational medicine rotation was developed that included focused clinical exposure to OEM patients and studies that would lead to eligibility for a certificate of additional qualification in occupational medicine. Learner evaluations included chart reviews and patient satisfaction surveys. Program evaluations included interviews with occupational health managers. The residents were judged by their preceptors to have performed well. The responses of the health managers and the patients were positive. This program in occupational medicine was found to be educationally sound with demonstrated community benefit and patient satisfaction. Further, it is cost-effective, requiring no external funding. PMID- 11381566 TI - Time-dependent oral absorption models. AB - The plasma concentration-time profiles following oral administration of drugs are often irregular and cannot be interpreted easily with conventional models based on first- or zero-order absorption kinetics and lag time. Six new models were developed using a time-dependent absorption rate coefficient, ka(t), wherein the time dependency was varied to account for the dynamic processes such as changes in fluid absorption or secretion, in absorption surface area, and in motility with time, in the gastrointestinal tract. In the present study, the plasma concentration profiles of propranolol obtained in human subjects following oral dosing were analyzed using the newly derived models based on mass balance and compared with the conventional models. Nonlinear regression analysis indicated that the conventional compartment model including lag time (CLAG model) could not predict the rapid initial increase in plasma concentration after dosing and the predicted Cmax values were much lower than that observed. On the other hand, all models with the time-dependent absorption rate coefficient, ka(t), were superior to the CLAG model in predicting plasma concentration profiles. Based on Akaike's Information Criterion (AIC), the fluid absorption model without lag time (FA model) exhibited the best overall fit to the data. The two-phase model including lag time, TPLAG model was also found to be a good model judging from the values of sum of squares. This model also described the irregular profiles of plasma concentration with time and frequently predicted Cmax values satisfactorily. A comparison of the absorption rate profiles also suggested that the TPLAG model is better at prediction of irregular absorption kinetics than the FA model. In conclusion, the incorporation of a time-dependent absorption rate coefficient ka(t) allows the prediction of nonlinear absorption characteristics in a more reliable manner. PMID- 11381567 TI - In vivo cerebral pharmacokinetics and pharmacodynamics of diazepam and midazolam after short intravenous infusion administration in sheep. AB - The cerebral pharmacokinetics and pharmacodynamics of midazolam and diazepam were examined in chronically instrumented sheep via measurements of their arterio venous concentration difference across the brain during and after 2-min i.v. infusions. Diazepam (30 mg) or midazolam (10 mg) were administered on 5 separate occasions to 4 sheep. For both drugs, rapid cerebral uptake occurred during the infusion, which quickly turned to elution in the postinfusion period. However, this process was more rapid for midazolam than diazepam. The cerebral pharmacokinetics of both was better described by a kinetic model with slight membrane limitation rather than flow limitation. For diazepam, the estimated brain:plasma partition coefficient was 2.67, and the first and second compartments filled with half-lives of 2.2 and 0.5 min, respectively. For midazolam, these values were 0.27, 0.26 and 1.34 min, respectively. In a subset of sheep, pulmonary arterial-arterial gradients were too small to measure suggesting limited metabolism and small distribution volumes for both drugs in the lungs. Simultaneous dynamic measurements of cerebral blood flow and algesimetry lagged behind both the arterial and sagittal sinus blood concentrations. The changes in cerebral blood flow were best described by a previously published a dynamic model that incorporated long half-lives for drug dissociation from the benzodiazepine receptor (13.3 and 5.5 min for midazolam and diazepam, respectively). Effect compartment modeling of the cerebral blood flow data showed apparent effect compartment half-lives (t1/2,keo) that were longer than the half-lives of cerebral equilibration. PMID- 11381568 TI - On sample size calculation in bioequivalence trials. AB - Sample size calculation plays an important role in bioequivalence trials. In practice, a bioequivalence study is usually conducted under a crossover design or a parallel design with raw data or log-transformed data. In this paper, we discuss the differences in sample size calculation between a crossover design and a parallel design with raw data or log-transformed data. Formulas for sample size calculation under a crossover design and a parallel design with raw data or log transformed data are derived. A brief discussion for the relationship among these formulas is given. PMID- 11381569 TI - Evaluating pharmacokinetic/pharmacodynamic models using the posterior predictive check. AB - The posterior predictive check (PPC) is a model evaluation tool. It assigns a value (pPPC) to the probability that the value of a given statistic computed from data arising under an analysis model is as or more extreme than the value computed from the real data themselves. If this probability is too small, the analysis model is regarded as invalid for the given statistic. Properties of the PPC for pharmacokinetic (PK) and pharmacodynamic (PD) model evaluation are examined herein for a particularly simple simulation setting: extensive sampling of a single individual's data arising from simple PK/PD and error models. To test the performance characteristics of the PPC, repeatedly, "real" data are simulated and for a variety of statistics, the PPC is applied to an analysis model, which may (null hypothesis) or may not (alternative hypothesis) be identical to the simulation model. Five models are used here: (PK1) mono-exponential with proportional error, (PK2) biexponential with proportional error, (PK2 epsilon) biexponential with additive error, (PD1) Emax model with additive error under the logit transform, and (PD2) sigmoid Emax model with additive error under the logit transform. Six simulation/analysis settings are studied. The first three, (PK1/PK1), (PK2/PK2), and (PD1/PD1) evaluate whether the PPC has appropriate type I error level, whereas the second three (PK2/PK1), (PK2 epsilon/PK2), and (PD2/PD1) evaluate whether the PPC has adequate power. For a set of 100 data sets simulated/analyzed under each model pair according to a stipulated extensive sampling design, the pPPC is computed for a number of statistics in three different ways (each way uses a different approximation to the posterior distribution on the model parameters). We find that in general; (i) The PPC is conservative under the null in the sense that for many statistics, prob(pPPC < or = alpha) < alpha for small alpha. With respect to such statistics, this means that useful models will rarely be regarded incorrectly as invalid. A high correlation of a statistic with the parameter estimates obtained from the same data used to compute the statistic (a measure of statistical "sufficiency") tends to identify the most conservative statistics. (ii) Power is not very great, at least for the alternative models we tested, and it is especially poor with "statistics" that are in part a function of parameters as well as data. Although there is a tendency for nonsufficient statistics (as we have measured this) to have greater power, this is by no means an infallible diagnostic. (iii) No clear advantage for one or another method of approximating the posterior distribution on model parameters is found. PMID- 11381571 TI - Supreme Court restricts ADA Title I as applied to the States. PMID- 11381570 TI - Is mixed effects modeling or naive pooled data analysis preferred for the interpretation of single sample per subject toxicokinetic data? AB - The purpose of this study was to evaluate whether mixed effects modeling (MEM) performs better than either noncompartmental or compartmental naive pooled data (NPD) analysis for the interpretation of single sample per subject pharmacokinetic (PK) data. Using PK parameters determined during a toxicokinetic study in rats, we simulated data sets that might emerge from similar experiments. Data sets were simulated with varying numbers of animals at each sampling time (4 48) and the number of samples taken (1-3) from each individual. Each data set was replicated 50 times and analyzed using several variations of MEM that differed in the assumptions made regarding intraindividual error, NPD, and a graphical noncompartmental method. These analyses attempted to retrieve the underlying parameter and covariate effect values. We compared these analysis methods with respect to how well the underlying values were retrieved. All analysis methods performed poorly with single sample per subject data but MEM gave less biased estimates under the simulated conditions used here. MEM performance increased when covariate effects were sought in the analysis compared with analyses seeking only PK parameters. Decreasing the number of animals used per sampling time from 48 to 16 did not influence the quality of parameter estimates but further reductions (< 16 animals per sampling time) resulted in a reduced proportion of acceptable estimates. Parameter estimate quality improved and worsened with MEM and NPD, respectively, when additional samples were obtained from each individual. Assumptions made regarding the magnitude of intraindividual error were unimportant with single sample per subject data but influenced parameter estimates if more samples were obtained from each individual. MEM is preferable to both NPD and noncompartmental approaches for the analysis of single sample per subject data but even with MEM estimates of clearance are often biased. PMID- 11381572 TI - Community integration of individuals with disabilities: an update on Olmstead litigation. PMID- 11381573 TI - [Metallic biomaterials for coronary stents]. AB - The introduction of coronary stents is a milestone in interventional cardiology. Two landmark studies have shown that stainless steel stents significantly decrease the restenosis rate as compared to balloon angioplasty. This fact led to a marked increase of stent implantation since the first stent implantation by Jacques Puel in 1986. Although the concept of coronary stenting significantly improved the interventional therapy of coronary artery disease, restenosis remains a major unsolved drawback of this technique. In addition to procedure and disease related factors like implantation pressure and plaque burden, data suggest that the stent as a medical implant plays a crucial role in the process of neointima formation. Since its introduction in cardiology, more than 50 different stents of different configuration and material have been developed. Although recent publications report of promising results using biodegradable materials, almost all coronary stents commercially available at the moment are made of metallic alloys. Whereas first generation stents were made exclusively from stainless steel and only minor interest was focussed on the stent material in the manufacture of coronary stents, recent studies strongly suggest that the metallic alloy used has a direct impact on the extent of neointima formation. Thus, metallic alloys differ not only with respect to mechanical features, but also by their biocompatible properties. These two factors are of major importance in the induction of vessel wall injury, inflammatory processes and cell proliferation. In the first part, the present paper reviews the metallurgic characteristics of metallic materials, which are currently used or under investigation in the production of coronary stents. In the second part, clinical and experimental results are summarized with respect to their biocompatibility and impact on the process of restenosis formation. PMID- 11381574 TI - [Dilated cardiomyopathy in childhood]. AB - In childhood, dilative cardiomyopathy (DCM) has a prevalence of 2.6 patients in 100,000 inhabitants. Manifestation age is, in 75%, the first two years of life. There are no specific symptoms. Diagnosis is often made, when congestive heart failure occurs. Despite intensive therapy with digitalis, diuretics and ACE inhibitors, DCM is the main indication for heart transplantation. The prognosis is critical: 11 studies with approximately 450 children showed a mean 1-year survival rate of 75% and a 5-year survival rate of 60%. In most cases etiology of DCM remains unclear ("idiopathic"). In the second position (approximately 40%) is inflammatory pathogenesis. While familiar DCM is manifested mainly during the 3rd and 4th decade, specific cardiomyopathy is often found during infancy: inborn errors of metabolism, neuromuscular diseases or malformation syndromes are the causes of a cardiomyopathy, which can appear as dilative or hypertrophic cardiomyopathy. A causal therapy exists only in a few cases. PMID- 11381575 TI - [Prognostic relevance of left ventricular diastolic function parameters in dilated cardiomyopathy]. AB - Patients with dilated cardiomyopathy (DCM) generally have an impaired functional capacity and poor long-term out-comes. A mortality of 5-15% per year has been described actually. Aim of this study was to verify the prognostic relevance of invasive and non-invasive parameters of diastolic function in patients with DCM. In 33 patients with DCM, cardiac catheterization was performed and left ventricular systolic (ejection fraction (EF; %)); left ventricular enddiastolic pressure (LVEDP; mmHg) and diastolic function (time constant of relaxation (T, ms); the constant of myocardial stiffness (b) were derived from biplane laevocardiography and simultaneous micromanometric registration of pressure volume curves. For evaluation of clinical out-come, the follow-up period was defined as beginning on the day after cardiac catheterization and ending on the most recent date or with a cardiac event (death or cardiac transplantation). All patients were reevaluated for NYHA functional class and completed a standard questionnaire. The following hemodynamic parameters were evaluated: invasive parameters of left ventricular diastolic function (constant of relaxation: tau (ms), constant of myocardial stiffness: b)), as well as parameters of systolic function (ejection fraction (EF; %)), left ventricular pressure (LVEDP; mmHg), left ventricular muscle mass index (LVMMI; g/m2), left ventricular enddiastolic volume index (LVEDVI; ml/m2) and non-invasive parameters of morphological data, left ventricular systolic (fractional shortening (FS, %) and ejection fraction) and diastolic parameters with echocardiography. During the follow-up period of 36 months, 11 of 33 patients experienced a major cardiac event (cardiac death n = 8, heart transplantation n = 3). The major cause of death was progressive pump failure. The remaining 22 patients were further classified with respect to changes in functional status. While clinical symptoms could be improved medically in patients with moderate increase of myocardial stiffness, patients with severe increase of myocardial stiffness (b: 76.1 +/- 12.1 vs 17.9 +/- +8.1, p < 0.001) could not be improved and suffered more cardiac events. Doppler echocardiographic measurements in these patients showed a restrictive filling pattern (VE 0.91 +/- 0.21 vs 0.64 +/- 0.18 m/s; p < 0.01; VA 0.52 +/- 0.23 vs 0.57 +/- 0.24 m/s; p < 0.01, deceleration time 129 +/- 17 vs 211 +/- 14 ms; p < 0.01). The medical heart failure therapy was comparable in both groups. In patients with cardiac events, the diastolic left ventricular variables did not significantly differ between patients who underwent heart transplantation and those who died. Patients who demonstrated a sole impairment of relaxation (tau: > 50 ms) suffered no cardiac events. Impaired diastolic function contributes to the clinical picture of congestive heart failure. Parameters of left ventricular diastolic function are powerful and important predictors of major cardiac events in patients with DCM, like heart transplantation and non-sudden death, and may indicate future clinical success of medical treatment. Invasive and non-invasive parameters of diastolic function reveal comparable information for the estimation of prognosis of patients with DCM in order to initiate early therapy. PMID- 11381576 TI - [Leptin: a parameter for metabolic changes in heart failure]. AB - OBJECTIVE: Advanced chronic heart failure is a hypercatabolic state with an imbalance between anabolic and catabolic metabolism and finally progressive loss of both muscle mass and adipose tissue. Leptin, the product of the obesity gene, is a hormone secreted by adipocytes. Therefore, we tested the hypothesis that plasma leptin concentrations are reduced in advanced chronic heart failure. METHODS: In 20 patients with chronic congestive heart failure (LVEF 23 +/- 6%) and 20 healthy controls (LVEF 65 +/- 8%) matched for gender, age, and body mass index, fasting plasma leptin (ELISA) and TNF alpha (ELISA) were measured. Follow up examination was performed after 1 year. RESULTS: The fasting plasma leptin concentrations of patients with NYHA grade III (8.4 +/- 3.8 ng/ml*) and NYHA grade IV (4.6 +/- 2.4 ng/ml dagger) were significantly lower as compared with the controls (11.2 +/- 3.1 ng/ml; *p < 0.05, dagger p < 0.01). In patients with NYHA grade II plasma leptin levels were significantly elevated as compared with the healthy controls (14.9 +/- 4.2 ng/ml). TNF alpha was higher in heart failure patients than in healthy controls (8.6 +/- 3.6 pg/ml; 5.9 +/- 2.1 pg/ml; respectively; p < 0.05), but did not correlate with the NYHA functional class. Mortality of the controls was 0%, whereas 15% (n = 3) in the congestive heart failure group; one patient (5%) needs an urgent heart transplantation. All of those patients had leptin concentrations below 5 ng/ml. CONCLUSIONS: Plasma leptin concentrations correlate with the NYHA functional class suggesting anabolic metabolism in NYHA class II and catabolic metabolism in advanced heart failure which might be of prognostic relevance. PMID- 11381577 TI - [The Ca(2+) sensitizers CGP 48506 and EMD 57033, but not the Na(+) channel modulator BDF 9148, prolong relaxation in isolated cardiomyocytes of the guinea pig]. AB - The sodium channel modulator DPI 201-106 has been described to posses Ca(2+) sensitizing properties. Therefore, the present study investigated the inotropic effect of the Na(+)-channel modulator BDF 9148 (1 microM), a congener of DPI 201 106, in comparison with the Ca(2+)-sensitizers CGP 48506 (1-50 mumol/l) and EMD 57033 (1-30 mumol/l) in electrically driven left ventricular cardiomyocytes isolated from guinea pigs. The changes of the contraction amplitude in comparison to the basal cell shortening (cell shortening in micron and %) were continuously recorded with a one-dimensional high speed camera. BDF 9148, CGP 48506, and EMD 57033 exerted a significant increase in the contraction amplitude (p < 0.05 vs. control). The maximal positive inotropic effects of CGP 48506 (50 mumol/l) and EMD 57033 (30 mumol/l) were +249 +/- 30% and +226 +/- 28%, respectively. The corresponding value for BDF 9148 (1 mumol/l) was +176 +/- 16%. However, only the Ca(2+)-sensitizers CGP 48506 and EMD 57033, but not BDF 9148, prolonged the contractile twitch. Especially in patients with an already enhanced intracellular myocardial Ca(2+)-concentration, Ca(2+)-sensitizers, which impair relaxation, may be disadvantageous for therapeutical use despite their positive inotropic effect. PMID- 11381578 TI - Reentry circuit location and left atrial three-dimensional activation patterns in left atrial flutter. AB - We report the case of a patient with left atrial flutter (LAF) in whom the reentry circuit location was mapped with a 64-electrode basket catheter deployed in the left atrium. Left atrial three-dimensional activation patterns were constructed with a software program and presented as color-coded isochrones. The reentrant activation traveled preferentially around the mitral annulus in a clockwise direction. It consisted of a single reentry confined anteriorly by the mitral annulus and posteriorly by an anatomic-functional barrier composed of a functional conduction block extending between pulmonary veins and surrounding a part of the posterior wall of the left atrium. The lower portion of the posterior wall and the anterior wall in close proximity to mitral annulus were preferentially used by the reentrant impulse. PMID- 11381579 TI - [Acute myocardial infarct in pheochromocytoma crisis. Early coronary angiography findings and echocardiography follow-up]. AB - A myocardial infarction is a rare complication of a pheochromocytoma. A pheochromo-cytoma crisis may occur spontaneously, during pregnancy, or may be induced by a local trauma of the tumor or by drugs. We present a case report of a 41-year-old woman without anamnestic episodes of hypertension or angina pectoris. During angiography of the mesenteric arteries for further diagnostics of a sonographically suspected liver tumor, she developed an acute pulmonary edema and a cardiogenic shock with the electro- and echocardiographic findings of a large anterolateral-apical-diaphragmal myocardial infarction. The immediate coronary angiography 90 min after the onset of the myocardial infarction showed normal coronary arteries with normal coronary blood flow of the arteries supplying myocardial areas with akinetic segments and those arteries supplying hyperkinetic segments. The blood catecholamine levels at this time were excessively elevated. The left ventricular function improved to almost normal within the next 4 weeks with the beginning of the improvement already before the abdominal tumor was surgically removed at day five. The histology documented a pheochromocytoma with acute necrosis. The early invasive findings support the hypothesis that a reversible spasm of several epicardial arteries and not a direct toxic effect of catecholamines could have been the cause of the small myocardial infarction and the observed large myocardial stunning. PMID- 11381580 TI - Local recurrence of soft tissue sarcoma. A Scandinavian Sarcoma Group Project. AB - The aim of this project was to investigate the diagnosis, treatment and consequences of local recurrence of soft tissue sarcoma (STS). It is based on patients reported to the Karolinska Hospital Sarcoma Register and the Scandinavian Sarcoma Group Register. Demographic and treatment data, based on 1613 adult patients reported to the Scandinavian Sarcoma Group Register by sarcoma centers in Norway, Sweden and Finland are presented. They all had STS of the extremities or trunk wall, and were diagnosed between 1986 and 1995. One third of the tumors were subcutaneous and two thirds deep-seated. The median size was 7 (1-35) cm and 75% were high grade. Metastases at presentation were diagnosed in 8% of the patients. Two thirds of the patients were referred to a sarcoma center before surgery. The preoperative morphologic diagnosis was made by fine-needle aspiration cytology in 72%. Among patients with final treatment for primary tumor at a sarcoma center (n = 1331), the surgical margins were wide or better in 76% of subcutaneous lesions, and in 58% of deep-seated lesions. Adjuvant radiotherapy has not generally been considered indicated after wide or compartmental excisions in Scandinavia. Overall, 23% of patients managed by surgery had adjuvant radiotherapy. Among patients with an intralesional or marginal excision, 44% had postoperative radiotherapy. Patients treated outside of sarcoma centers were seldom referred for radiotherapy. The crude local recurrence rate was 225/1331 (17%) among the patients with final treatment for primary tumor at a sarcoma center. The local recurrence rate after local surgery for high-malignant deep-seated STS was 103/391 (26%). The rate was 25/64 (39%) after an intralesional/marginal margin without postoperative radiotherapy versus 28/119 (24%) when radiotherapy was given. Fine-needle aspiration cytology (FNAC) was used to diagnose suspected local recurrences. 95 FNAC were performed in 86 patients from Karolinska Hospital. There were 47 local recurrences, of which 44 were diagnosed correctly by FNAC; one biopsy was inconclusive, and two lesions were incorrectly assessed as benign. 39 patients proved to have benign lesions in the scar examined cytologically on 50 occasions. None of the specimens was regarded as malignant, but in 4 cases FNAC was inconclusive. The inconclusive or false cytological diagnoses had no serious clinical consequences. Among 205 patients with local recurrence identified in the SSG Register 1987-1995, 169 patients were surgically treated. An intralesional or marginal margin was achieved in 110 of these patients, 59 of whom were also given radiotherapy. 54 of the 169 patients had a second local recurrence. The second local recurrence rate was 0.50 if the first local recurrence was treated using surgery with a marginal margin alone, compared to 0.28 if treated using either surgery with a marginal margin and radiotherapy, or a wide margin (p = 0.0008). In extremity STS, the amputation rate for local recurrences was 0.22, compared to 0.09 for primary tumors. The overall 5-year MFS was 0.72 (95% CI 0.68-0.76). High histopathological malignancy grade (Relative Risk 3.0; 95% CI 1.5-6.3) and an inadequate surgical margin (2.9; 95% CI 1.8-4.6) were independent risk factors for local recurrence. High histopathological malignancy grade and large tumor size (> 7 cm) were the most important risk factors for metastasis. Local recurrence was associated with an increased risk of metastasis (4.4; 95% CI 2.9 6.8), but an inadequate surgical margin was not a risk factor for metastasis (1.1; 95% CI 0.8-1.7). In conclusion, it is unlikely that local recurrence of STS is a major source of metastases. It nevertheless represents a costly, complicated and emotionally difficult problem. More radical surgical margins would improve the local recurrence rate, but this can hardly be achieved for center-operated patients without increasing the amputation rate. Instead, local control will improve by giving radiotherapy to all patients after marginal surgery, and to selected patients with wide margins. Radiotherapy is indicated especially after a previous open biopsy or when a local recurrence might lead to an amputation. Furthermore, radiotherapy seems indicated after local recurrence, regardless of margin or grade. The most effective way of reducing costs and detriment associated with local recurrence is to increase referral to sarcoma centers before biopsy or surgery as primary surgical margins would then improve. PMID- 11381581 TI - Knee joint kinematics, fixation and function related to joint area design in total knee arthroplasty. AB - The aim was to study the influence of different designs of the joint area on tibial component fixation, kinematics and clinical outcome after a cemented total knee arthroplasty (TKA). The HSS score and a special questionnaire were used at the clinical examination. Conventional radiography was done to record the positioning of the implants and development of radiolucencies. The migration and inducible displacement were evaluated using radiostereometry (RSA). The kinematics of the knee during active extension was studied using dynamic RSA. In randomised and prospective studies 87 knees in 83 patients (28 male, 55 female, age 69, range 50-83) received an AMK (DePuy, Johnson & Johnson) TKA. The patients were divided into two groups. In group 1 the patients had varus/valgus deformities of < or = 5 degrees and the PCL was retained. The PCL was resected in group 2 where the patients had deformities exceeding 5 degrees and/or fixed flexion deformities of more than 10 degrees. In group 1 a flat (F, n = 20) or a concave (C, n = 20) design was implanted (study 3). In group 2 (study 4) the patients received a concave (n = 25) or a posterior-stabilised (PS, n = 22) tibial plateau. The migration of the tibial component, positioning of the prosthesis, development of radiolucencies and the clinical outcome was evaluated after 1 and 2 years. Twenty-two patients (11 F, 11 C) in group 1 (study 1) and 22 knees in 20 patients in group 2 (study 2, 11 C, 11 PS) were examined 1 year post operatively to evaluate the kinematics of the knee. Eleven normals served as controls. During active extension of the knee the inducible displacements of the tibial component were recorded in 16 knees (15 patients). Based on successful RSA examinations 5 knees (4 F, 1 C) from group 1 and 11 knees (5 C, 6 PS) from group 2 were selected (study 5). Abnormal kinematics and especially increased AP translations compared to normals (p < 0.0005) were recorded in all designs. The concave design showed the widest AP-translations in both studies. The clinical outcome in terms of HSS score did not differ between the flat versus the concave designs in study 1 and between the concave versus the PS implants in study 2. Up to two years the migration of the tibial component and the development of radiolucent lines were of the same magnitude for the flat versus the concave inserts in study 3 and the concave versus the PS design in study 4. Also did the positioning of the implant and the fulfillment of the patients expectations on the surgery preoperatively not differ. The AMK prosthesis migrated at about the same amount as have been reported for similar designs. In study 5 all implants showed a correlation between some of the inducible displacements (anterior posterior tilting and maximum total point motion) and the corresponding migration 0-2 years. The more the anterior tilt the more the migration in the same direction. If the PCL was sacrificed during the knee replacement the change into increased anterior tilt occurred earlier (i.e. at more degree of flexion) if a concave insert was used compared to the PS design. When the active extension reached 25 degrees there were more anterior tilt of the tibial component in the concave design (p = 0.001) and if the tibial plateau centre had a medial position (p < 0.0005). Compared with normal knees all prosthetic designs showed abnormal pattern of motion. The extent of this abnormality was influenced by the design of the joint area. A corresponding influence on the fixation of the tibial component could not be verified. The choice of joint area and recorded kinematics had no or small influence on the clinical results. Feelings of instability could to some extent be related to the kinematics of the knee joint. PMID- 11381582 TI - Adult child survivor syndrome on deprived childhoods of aging Holocaust survivors. AB - The present paper aims at understanding the psycho-(patho-)logical status of aging child survivors (CSs) of the Holocaust. BACKGROUND: CSs had deprived childhoods, were exposed to massive trauma and losses, then rebuilt their existences. Their post-Holocaust malaise was suppressed and split off from daily life and overt activities, only to reappear, after a delay, in different forms and intensities, further aggravating upon aging. METHOD: Following a definition of age, origin, number of living CSs, types of early traumatic impacts, and introductory remarks on what CSs experienced as children and as young adults in the postwar society, the psychosocial research on adult and aging CSs is reviewed. RESULTS: The concept of childhood-deprivation and its late sequelae can be substantiated. Definitions of child-survivor complex, CS syndrome and adult child-survivor syndrome are proposed. Non-professional caretakers discerned earlier the existence or extent of the adult CS syndrome Limitations of psychiatric investigation of ACs are the lack of agreed-upon diagnostic criteria. PMID- 11381583 TI - Narrative group therapy with aging child survivors of the Holocaust. AB - Child survivors of the Holocaust, aged 1-16 years at liberation in 1945, now well beyond mid-life, are approaching the gerontic stage of their psychosocial development. They find themselves confronted with the demands of aging, as well as unmet needs related to their damaged early development. AMCHA, the National Israeli Center for Psychosocial Support of Holocaust Survivors and the Second Generation, attempted to create a framework for exploring and meeting these needs. An account is given of a narrative group consisting of aging, non-clinical child survivors who met once monthly over a period of more than five years. The individual narratives spun out within the group meetings weave a common texture which served as a context for mutual recognition, dialogue and restoration of meaning to fragmented childhood memories. Retrieval of the lost life stories served as a bridge for the belated "homecoming" of the "lost children." One of the problems is how to resolve the group with the right amount of tears because, regardless of the achievements, mourning is not completed. PMID- 11381584 TI - Child survivors of the Holocaust: symptoms and coping after fifty years. AB - Holocaust survivors who were children during WW II have now reached the age of 52 to 67. Until about 10 years ago their voices were barely heard in society. Their successful adaptation to life may have contributed to this invisibility. However, reaching this stage of life, which is associated with the need to review life and with the crises of retirement and renewed losses, has activated the survivors to deal with their childhood. The impossibility of avoiding traumatic memories and an urge to deal with them have also contributed to the societal process of the survivors organizing and speaking out. Very little is known about this group with regard to their mental health status and the way they cope with their childhood memories. The present controlled double-blind study uses a randomized nonclinical sample and focuses on the level of psychosocial and post-traumatic symptoms, on achievement motivation, and on the way child survivors perceive the surrounding world. The results indicate a slightly higher level of psychosocial symptoms in the child survivors group (CS) than in the control group, a high level of post traumatic symptomatology, and achievement motivation based mainly on the fear of failure. Surprisingly, the child survivors group shows a pattern of more positive views of the world than does the control group. This can be understood as a greater need to compensate for the lack of security suffered in childhood by creating a meaningful world in a chaotic reality. PMID- 11381585 TI - Psychopathology in children of Holocaust survivors: a review of the research literature. AB - The literature on transgenerational transmission of Holocaust trauma has grown into a rich body of unique psychological knowledge with almost 400 publications. For the time being, however, the transgenerational effect of the Holocaust on the offspring remains a subject of considerable controversy. The main question involves the presence or absence of specific psychopathology in this population. Psychotherapists kept reporting various characteristic signs of distress while research failed to find significant differences between offspring and comparative groups. In an effort to settle this question, the present review of the research literature provides a summary of the findings of 35 comparative studies on the mental state of offspring of Holocaust survivors, published between 1973-1999. This extensive research indicates rather conclusively that the non-clinical population of children of Holocaust survivors does not show signs of more psychopathology than others do. Children of Holocaust survivors tend to function rather well in terms of manifest psychopathology and differences in the mental state of offspring and people in general are small according to most research. The clinical population of offspring, however, tend to present a specific "psychological profile" that includes a predisposition to PTSD, various difficulties in separation-individuation and a contradictory mix of resilience and vulnerability when coping with stress. PMID- 11381586 TI - A controlled double-blind study on children of Holocaust survivors. AB - The effects of the Holocaust on the offspring of survivors have remained unclear in discussions between clinicians, clinical researchers and empirical researchers. The authors report on a controlled double-blind study designed to test these effects using the sensitivity of clinicians to intrapsychic constellations and processes. The all female sample was randomly selected from several Jerusalem neighborhoods. The index subjects (n = 31), born between 1946 and 1960, had at least one parent (mother) who suffered persecution at the hands of the Nazi regime during WWII. The controls (N = 31), matched for age, educational status and birth order, were born to parents who had not lived under the occupation of the Nazi regime and had not suffered losses of close relatives in WWII. In-depth double-blind interviews, conducted by experienced psychodynamic psychotherapists, focused on personality characteristics without questioning individual development. The only measure used was a 48-item questionnaire completed by the therapists at the end of the interview. The study showed that daughters of Holocaust survivors are characterized by more problems in the realm of separation individuation issues. It also confirmed previous findings that the offspring of Holocaust survivors do not show more psychopathology than the general population. PMID- 11381587 TI - Perceived parental rearing behavior in children of Holocaust survivors. AB - Holocaust survivors have often been described as inadequate parents. Their multiple losses were assumed to create child-rearing problems around both attachment and detachment. Empirical research, however, has yielded contradictory evidence regarding the parenting behavior of Holocaust survivors when investigated with classical parenting instruments. The present pilot-study investigated parental behavior with a new self-report instrument that also included salient Holocaust dimensions. The parent perception of 159 adult children of Holocaust survivors was thus compared with 151 control subjects. Factor analysis of data yielded four major kinds of parental rearing behaviors: transmission; affection; punishing and over-protection. While the second generation group rated their parents higher on transmission, other differences in child-rearing practices were small, if taken as a whole. These findings largely support the descriptive literature on transgenerational transmission of trauma while at the same time refuting the view that Holocaust survivors function more inadequately than other parents do. PMID- 11381588 TI - Isolation and subfractionation of mitochondria from animal cells and tissue culture lines. PMID- 11381589 TI - In vivo measurements of respiration control by cytochrome c oxidase and in situ analysis of oxidative phosphorylation. PMID- 11381590 TI - Assay of mitochondrial ATP synthesis in animal cells. PMID- 11381591 TI - Measurement of membrane permeability and permeability transition of mitochondria. PMID- 11381592 TI - Assaying actin-binding activity of mitochondria in yeast. PMID- 11381593 TI - Analysis and prediction of mitochondrial targeting peptides. PMID- 11381594 TI - Assaying protein import into mitochondria. PMID- 11381595 TI - Analysis of protein-protein interactions in mitochondria by coimmunoprecipitation and chemical cross-linking. AB - Many different techniques have been employed to analyze protein-protein interactions. Coimmunoprecipitation and chemical cross-linking have been used extensively to study mitochondrial biogenesis. Both techniques have proven to be powerful methods to investigate the sequential interactions of precursor proteins with the various components of the translocation machineries in the mitochondrial membranes. Similarly, protein-protein interactions during processes such as protein synthesis, folding, and degradation can be studied. Moreover, the composition of the oligomeric protein complexes of mitochondria, such as respiratory chain complexes or protein translocation machineries, can be determined. The general principles and protocols of these methods are described and illustrated with typical examples. PMID- 11381596 TI - Blue-native gels to isolate protein complexes from mitochondria. PMID- 11381597 TI - Application of electron tomography to mitochondrial research. PMID- 11381598 TI - Epitope tagging and visualization of nuclear-encoded mitochondrial proteins in yeast. PMID- 11381599 TI - Targeting of green fluorescent protein to mitochondria. PMID- 11381600 TI - Assessment of mitochondrial membrane potential in situ using single potentiometric dyes and a novel fluorescence resonance energy transfer technique. PMID- 11381601 TI - Optical imaging techniques (histochemical, immunohistochemical, and in situ hybridization staining methods) to visualize mitochondria. PMID- 11381602 TI - Visualization of mitochondrial movement in yeast. PMID- 11381604 TI - Isolation and subfractionation of mitochondria from the yeast Saccharomyces cerevisiae. PMID- 11381603 TI - Targeting of reporter molecules to mitochondria to measure calcium, ATP, and pH. PMID- 11381605 TI - Genetic transformation of Saccharomyces cerevisiae mitochondria. PMID- 11381606 TI - Transmitochondrial technology in animal cells. PMID- 11381607 TI - Diagnostic assays for defects in mitochondrial DNA replication and transcription in yeast and human cells. PMID- 11381608 TI - Analysis of mitochondrial translation products in vivo and in organello in yeast. PMID- 11381609 TI - Numerical methods for handling uncertainty in microarray data: an example analyzing perturbed mitochondrial function in yeast. PMID- 11381610 TI - Isolation and subfractionation of mitochondria from plants. PMID- 11381611 TI - Assessing functional integrity of mitochondria in vitro and in vivo. PMID- 11381612 TI - Assaying mitochondrial respiratory complex activity in mitochondria isolated from human cells and tissues. PMID- 11381613 TI - Establishment of specific pathogen-free guinea-pig colonies using limited-flora guinea-pigs associated with conventional guinea-pig flora, and monitoring of their cecal flora. AB - Six groups of limited flora (LF) Hartley guinea-pigs were produced by inoculation of hysterectomy-derived GF guinea-pigs with various combinations of cecal bacteria of conventional (CV) guinea-pigs to determine the effective bacterial cocktails for the establishment of a specific pathogen free (SPF) colony. Bifidobacterium magnum (Bif) isolated from CV guinea-pigs was used for pretreatment. The mortality of LF guinea-pigs inoculated with only Bif was 75%, and that of those inoculated with Bif plus chloroform-treated cecal suspension (CHF) or Bif plus CHF plus 32 isolates from CV guinea-pigs was 40 to 66.7%. These three groups were in an unhealthy condition with mucoid enteritis-like diarrhea. However, the mortality of LF guinea-pigs inoculated with the anaerobic growth on EG plates injected with 10(-5) dilution of cecal contents (CF) or inoculated with Bif plus CF was 6.3 and 15%, respectively. These latter two groups of LF guinea pigs were transferred to separate barrier rooms and some of the LF guinea-pigs were maintained in isolators as a source of intestinal flora for SPF guinea-pigs. The composition of cecal flora of LF guinea-pigs was stable for a long time, and bacteroidaceae and peptococcaceae were maintained as predominant components. The basic composition of the cecal flora of SPF guinea-pigs originated from LF guinea pigs, which consists mainly of the anaerobic bacteria, was not changed over a long period, and the flora composition became similar to that in CV guinea-pigs. Guinea-pig-specific pathogens from the SPF colonies were not detected during experiments. PMID- 11381614 TI - Identification of epistatic interactions involved in non-insulin-dependent diabetes mellitus in the Otsuka Long-Evans Tokushima Fatty rat. AB - The Otsuka Long-Evans Tokushima Fatty (OLETF) rat is an animal model for obese type non-insulin-dependent diabetes mellitus (NIDDM) in humans. Our present investigation was designed to identify epistatic interactions influencing NIDDM by performing least squares analysis of variance of all pairs of informative markers in 160 F2 progenies bred from an intercross of OLETF and Fischer-344 rats. We identified four interactions between Nidd15/of (chromosome 7) and Nidd16/of (chromosome 14), Nidd15/of and Nidd17/of (chromosome 15), Nidd16/of and Nidd18/of (chromosome 15), and Nidd16/of and Nidd19/of (chromosome 17), which account for a total of approximately 40% of the genetic variation of entire glucose levels after glucose challenge in the F2. The Nidd16/of locus, which is involved in three of four digenic interactions, and the Nidd19/of are likely to correspond to Nidd2/of and Nidd14/of, NIDDM loci previously identified in the F2 by single-QTL model and multiple-QTL model, respectively, while Nidd15/of, Nidd17/of and Nidd18/of loci reflect novel NIDDM loci. An aberrant increase of the entire glucose level due to synergism occurs in the double OLETF homozygote genotype of Nidd15/of and Nidd16/of, and of Nidd16/of and Nidd19/of, as well as in the OLETF homozygote genotypes of Nidd15/of and Nidd16/of, respectively, combined with the heterozygote genotypes of Nidd17/of and Nidd18/of. These findings demonstrate that inter-allelic interactions are likely to be an important component of NIDDM susceptibility. PMID- 11381615 TI - A comparative study on the integration of exogenous DNA into mouse, rat, rabbit, and pig genomes. AB - Transgenic mammals, from small laboratory rodents to domestic animals, have been successfully produced to date, but their production efficiency within or across species has been variable. This is probably due to the differences in the type of injected DNA and/or technical procedures employed in each laboratory, as well as the reproductive characteristics of the species. Here we report the direct comparison of the efficiencies of producing transgenic mice, rats, rabbits and pigs by one technician using a fusion gene composed of the bovine alpha S1-casein promoter and human growth hormone (hGH) gene. Before the fusion gene was injected into the zygotes, high magnitude centrifugation to visualize the pronuclei was necessary for all of the pig zygotes and one-third of the rabbit zygotes, but not for mouse and rat zygotes. Post-injection survival of the mouse zygotes (67.1%) was lower than those of the rat, rabbit and pig zygotes (89.6 to 100%). The volume change of the pronucleus following DNA injection was the lowest in mice (50% increase), moderate in rabbits (148% increase), and the most prominent in rats (238% increase). The data from only 1 pig zygote indicated a 22% increase in the pronucleus volume by DNA injection. The PCR analyses of the tail DNA of new born offspring indicated that 0.8% (4/493), 4.8% (22/463), 0.8% (3/367) and 0.9% (2/221) of the injected eggs in mice, rats, rabbits and pigs, respectively, developed into transgenic offspring. Some of the founder animals in all four species expressed the transgene in the mammary gland which was confirmed in hGH mRNA by RT-PCR and/or hGH peptide in Witch's milk with ELISA. These results suggest that the maximum volume of DNA solution injectable into the pronucleus is a possible factor explaining the species differences in the production of transgenic animals. PMID- 11381616 TI - Analysis of histochemically and morphometrically in the anterior belly digastric muscle of osteopetrotic (op/op) mice. AB - The fibers of the anterior belly digastric muscle of mice, fed a granulated diet for various periods, have been studied histochemically and morphometrically. The diameters of the anterior belly digastric fibers in normal mice fed only a granulated diet were smaller than those in mice fed a solid diet. Differences in the succinate dehydrogenase (SDH) activity of muscle fibers between op/op and normal mice gradually appeared in the anterior belly digastric muscle and, by the age of 90 days, under-development of muscle fibers was observed in the mild-belly region of the anterior belly digastric muscle of op/op mice fed a granulated diet. These results indicate mechanical stress in mastication plays an important role in the development of the anterior belly digastric muscle structures. PMID- 11381618 TI - X-ray hypersensitivity in the LEA rat: genetic linkage analysis of responsible loci. AB - The LEA rat was established from a Long-Evans rat closed colony as the control strain of the LEC rat, which is reported to exhibit several mutant phenotypes such as hepatic disorder (hts), blockage of the T cell differentiation (thid) and X-ray hypersensitivity (xhs1 and xhs2). Here we report that the LEA rat is hypersensitive to X-rays to a similar degree as the LEC rat, although it is normal with respect to the hts and thid phenotypes. We further performed genetic linkage analysis of X-ray hypersensitivity in the LEA rat. The quantitative trait loci (QTL) linkage analysis revealed that xhs1 but not xhs2 is the locus responsible for X-ray hypersensitivity in the LEA rat. PMID- 11381617 TI - In vitro migratory responses of swine neutrophils to actinobacillus pleuropneumoniae. AB - Swine neutrophils were quantitatively examined for the direct and indirect migratory responses to Actinobacillus pleuropneumoniae (APP) in vitro and the effects of pseudorabies virus (PrV), frequently co-infecting with APP, were also observed. About 30% of swine neutrophils responded to viable APP, while 3.2% of the neutrophils responded to 0.1% casein which served as the control. The migration of APP was not affected by preincubation of neutrophils with PrV, which inhibited the random migration. When the random migration was normalized to 1, the chemotactic indices for APP, opsonized-APP and casein were 64, 70 and 8.5, respectively. Heat-killed APP or E. coli lipopolysaccharide stimulated the production of interleukin-8 activity by adherent peripheral blood mononuclear cells (PBMC). Preincubation of PBMC with PrV inhibited the production of neutrophil attractant activity when stimulated with heat-killed APP. The results suggested that the direct chemotaxis of neutrophils to viable APP might contribute to early infiltration in Actinobacillus pleuropneumonia, and that PrV might inhibit indirect recruitment of neutrophils to infected lungs by compromising the functions of PBMC. PMID- 11381619 TI - Distribution and properties of rat intestinal alkaline phosphatase isoenzymes. AB - The ALP activities and properties of rat intestine cut into 20 segments were examined, and we were able to demonstrate that the ALP activity of upper intestine is high compared to that of lower intestine. This result coincided with those of other reports. However, we newly clarified that there is an ALP isoenzyme found in the lower intestine which can be inhibited by L-homoarginine. The molecular weight of the ALP isoenzyme was 136 kDa. In addition, it was clarified that there are several isoenzymes from upper to lower intestine. This study demonstrates that there exist isoenzymes, which are inhibited by L-HArg, in the intestine which are similar to the isoenzymes in the liver, bone and kidney. PMID- 11381621 TI - Two inbred strains of mice with high and low mammary tumor potentials established from the same basal stock of Swiss albino (SHN and SLN): history of mammary tumorigenesis and reproduction after the 30th generation of full-sib mating. AB - We established SHN and SLN strains of mice with a high and a low mammary tumor potentials from the same basal stock of Swiss albino. The selection history of females (mothers) for subsequent generations, mammary tumorigenesis and reproduction up to recent generations are presented and compared with the results up to F30 published in 1979. The changes with generation of each parameter varied in both strains except the conditions of female (mother) selection for subsequent generations, which were generally constant. However, the fluctuations in parameters up to F30 became generally smaller with advancing generations. Based on the fluctuations from generation to generation of mammary tumorigenic and reproductive parameters, an interdependency between mammary tumor potential and reproductivity is suggested in SLN, but less so in SHN. The pup growth rate was increased and associated with selection for mammary tumorigenesis in SHN; this parameter is usually used as an indicator of mother lactational performance in mice. The significance of selection for mammary tumor potential in association with reproductivity was further discussed. PMID- 11381620 TI - Age differences in immunohistochemical localizations of large proteoglycan, PG M/versican, and small proteoglycan, decorin, in the dermis of rats. AB - Proteoglycans were localized immunohistochemically in the dermis of Donryu rats, using monoclonal antibodies raised against large proteoglycan (PG-M/versican) and small proteoglycan (decorin). The localizations of these proteoglycans in the dermis were compared between young rats (22-day old) and old ones (24 or 30 months of age), and distinct age differences were observed. In the young dermis, PG-M/versican was observed to be abundant in almost all fibroblastic cells (both cytoplasm and cell processes) whose cellularity was very rich compared with the dermis of old rats. Decorin was only faintly visible in the interstitial fibrous elements of young dermis. In the old dermis, however, decorin was distinctly detected on the fibrous elements, which were diffusely distributed as a fibrous network, and likewise PG-M/versican was visible in only a few fibrous elements which seemed to be the fine processes of fibroblastic cells. In the border layer between epidermis and dermis as well as the basal layer surrounding hair follicles, both large and small proteoglycans could be observed in old dermis. Since decorin, which was abundant in old dermis, has been found to have a growth inhibitory effect, it is conceivable that decorin may be one of the Cell Growth Inhibitory Factors in aging as proposed by Tauchi et al. [17, 18]. PMID- 11381622 TI - Effects of olive, corn, sesame or peanut oil on the body weights and reproductive organ weights of immature male and female rats. AB - Olive, corn, sesame or peanut oil which have been used as vehicles in the immature rat uterotrophic assay or Hershberger assay, for detection of endocrine disrupting effects of environmental chemicals, was administered to ten immature female rats by subcutaneous injection from postnatal day (PND) 21 for 3 or 7 days, and each oil was also administered to ten male rats from PND 21 for 7 and 10 days. The body weights, and the weights of sex and sex accessory organs in female and male rats were measured. There were no significant differences in body weights of female rats between each oil group and the control group, while the body weight of male rats in the group given peanut or olive oil was significantly increased from 8 or 9 days after administration. There were no changes in the sex and sex accessory organ weights of female or male rats related to the endocrine disrupters. The results of the body weights and organ weights demonstrate that each oil is a suitable vehicle for the immature rat uterotrophic assay. However, each oil is suggested to be unsuitable for the Hershberger assay, because the analysis of changes of sex accessory organ weights in this assay might be confused by the increased body weights. PMID- 11381623 TI - Histological characteristics of the pelage skin of rough fur mice (C3H/HeJ- ruf/ruf). AB - Pelage skin of C3H/HeJ mice homozygous at an autosomal recessive mutant locus, rough fur (ruf) which is located on chromosome 9, was histologically analyzed. Sebaceous glands synthesizing lipids were larger in the mutant mice than in controls in an examination by Sudan IV staining. Electron microscopic analysis of the sebaceous gland showed that lipid droplets were denser in mutant mice than in control mice, and that they were irregular in shape in ruf mice while those of controls were round. Our results suggested that rough fur (ruf) mice might be an animal model for hyperlipogenesis of the pelage skin. PMID- 11381624 TI - Comparison of bacteriological, genetic and pathological characters between Escherichia coli O115a,c:K(B) and Citrobacter rodentium. AB - Murine pathogenic Escherichia coli O115a,c:K(B) (MPEC) is the causative agent of mouse megaenteron, the pathology of which resembles that of transmissible murine colonic hyperplasia caused by Citrobacter rodentium. We compared their genetic and pathological features to reveal the relationship between these two bacteria. To evaluate the genetic distances, 16S rDNA genes were sequenced and biochemical reactions were tested. Mouse strain susceptibility tests, using CF1 MPEC susceptible germfree mice and BALB/cA(Jic) resistant mice were performed. MPEC strains and C. rodentium showed more than 99.6% identity by comparison of 16S rDNA gene sequences. All results from biochemical reactions and the mouse strain susceptibility tests were identical. It is proposed that MPEC should be reclassified as C. rodentium. PMID- 11381625 TI - High rate of IgE-mediated histamine release from rat mesenteric mast cells. AB - IgE-dependent histamine release from rat mesenteric mast cells was investigated. Excised mesenterium was cut into pieces and incubated with IgE overnight at 4 degrees C for sensitization. Over 10 pieces of mesenterium specimen could be prepared from a rat. Antigen-induced histamine release from mesenterium specimen was initiated rapidly and reached a plateau in 5 min. In an optimal condition, over 50% of total histamine was released. In contrast, unpurified and purified peritoneal mast cells released only 22.5% and 5.3% of total histamine, respectively, upon IgE stimulation. Tranilast, a mast cell stabilizer, inhibited the histamine release from mesenteric mast cells significantly. The mesenterium might be useful material for studying tissue-associated mast cell activation. PMID- 11381626 TI - Experience of vein grafting in Gottingen minipigs. AB - We experimented with vein grafting surgery on Gottingen minipigs. Using the internal jugular vein for the tissue graft, we performed side-to-side anastomosis to the carotid artery, to which it runs parallel. One key point in this surgery was to prevent vasospasm of the carotid artery so as to keep the lumen sufficiently patent during anastomosis. The histopathological findings in the grafts which remained patent resembled those of vein grafts in humans. We therefore considered that this technique in minipigs can be applied for the study of coronary artery bypass surgery in humans. PMID- 11381627 TI - Spontaneous neoplastic lesions in aged Sprague-Dawley rats. AB - Neoplastic lesions were observed in untreated aged Sprague Dawley (SD) rats throughout their lifespan starting at 5 weeks. Their mean survival times were 89 to 105 weeks of age. The total tumor incidences were 70 to 76.7% and 87 to 95.8% in males and females, respectively. The common neoplasmas were pituitary adenoma and adrenal pheochromocytoma in both sexes, testicular Leydig cell tumor in males and mammary gland tumors, thyroidal C-cell adenoma and uterine stromal polyp in females. PMID- 11381628 TI - Neisseria meningitidis with decreased susceptibility to penicillin in Ontario, Canada 1997-2000. PMID- 11381629 TI - Blastocystis hominis: a new pathogen in day-care centres? PMID- 11381630 TI - [Prevalence and risk factors of root caries. A study of residents over 70 years old in the homes for the aged]. PMID- 11381631 TI - [Marginal periodontitis and cardiovascular diseases]. PMID- 11381632 TI - [Status of infectiology in Germany]. PMID- 11381633 TI - [Infections caused by Stenotrophomonas maltophilia (SMA) in intensive care patients--a prospective case-control study]. AB - BACKGROUND AND OBJECTIVE: The importance of Stenotrophomonas maltophilia (SMA) as an etiologic frequently polyresistant pathogen in severe nosocomial infections has increased. METHODS: In our prospective study we evaluated the risk factors of nosocomial infections by SMA in our internal intensive care unit (ICU) over a one year period from July 1997 to June 1998. RESULTS: 111 patients (80 men, 31 women, mean age +/- SD: 58.0 +/- 13.3 years) were treated for more than 5 days in the ICU. SMA were cultured in 16/111 patients (13 men, three women, mean age 57.8 +/- 3.4 years) out of bronchial secretions (68%), sputum (19%) and pleural fluid (13%). Univariate analysis resulted in 15 different risk factors (p < 0.05); however, multivariate analysis provided three independent risk factors: chronic obstructive pulmonary disease (OR 95% CI [1.91; infinity]), length of stay in the ICU (OR 95% CI [1.07; 1.26]) and therapy with carbapenems before admittance to ICU (OR 95% CI [0.56; 153]). Four of 16 patients died due to an SMA-infection, two by purulent exacerbations of a chronic bronchitis and two by sepsis. Molecular typing of 18 SMA isolates in 15 patients resulted in 9 different genetic types and a clonal dissemination could only be confirmed in three patients. CONCLUSIONS: In respiratory ICU SMA infections are favored by severe COPD, length of stay in the ICU and by selection pressure of applicated antibiotics, especially carbapenems. PMID- 11381634 TI - [Nosocomial epidemiology and transmission of Clostridium difficile infection]. AB - BACKGROUND: Clostridium difficile is of growing importance as a hospital-acquired pathogen. Pseudomembraneous colitis is the main clinical disease. Transmission and epidemiological features are not yet fully understood. PATIENTS AND METHODS: Stool samples from 1164 individuals (571 women and 593 men) attending were examined for the presence of C. difficile. Follow-up examinations and molecular typing methods were used for the detection nosocomial transmissions. Additionally, hospital-borne environmental samples as well as staff samples were tested. RESULTS: Incidence of C. difficile infection was 8.4%. Nearly all patients (92.9%) had antibiotics given. Using molecular typing nosocomial transmission was evident. Though, environmental samples in general had a low positivity, toilet chairs were contaminated in 15.4% and may be a potential source of transmission. Staff was positive in only one case. CONCLUSIONS: Prevention of infections with C. difficile becomes to be a major threat for the clinical and hygienic management. PMID- 11381636 TI - [Frequent infectious enteritis--1: Diagnosis]. PMID- 11381635 TI - [Fever and exanthema after trip to the Mediterranean area--Mediterranean spotted fever (Boutonneuse fever)]. AB - HISTORY AND CLINICAL FINDINGS: In connection with a period of residence in the Mediterranean basin (Portugal, Spain, Sicily) one woman and two men (age: 28, 31 and 40 years) got sick with fever in August and in the midst of October. In addition, they complained about headache in two cases and myalgias, arthralgia and vomiting in one case, respectively. Two of the vacationers (cases 2 and 3) had been accompanied by their dogs. One of them (case 2) was bitten of a tick. The woman (case 3) removed few days after vacation-beginning ticks from her dogs. Patients 1 and 2 appeared severely ill and presented with high fever and generalized maculopapular rash which included also the soles and palms. Patients 2 and 3 had a primarily-lesion ("eschar"). In patient 3 the eschar was pustular and associated with regional lymphadenitis. INVESTIGATIONS: All three patients showed an increased erythrocyte sedimentation rate and elevated liver enzymes. Cases 1 and 2 presented with a significantly elevated activity of lactate dehydrogenase, leukocyturia and microhematuria. Indirect immunofluorescent assay for specific antibodies directed against Rickettsia conorii revealed titers between 1:40 and 1:640 (normal: negative). DIAGNOSIS, TREATMENT AND COURSE: Diagnosis was based on the triad fever, rash and eschar as well as on epidemiological data which include a recent period of residence in the Mediterranean basin during the summer and contact with a dog. In case 1 diagnosis was difficult because of the absence of an eschar. This patient had been treated with penicillin for two days without success. Only the parenteral administration of ciprofloxacin caused complete defervescence and clinical improvement within two days. A complete defervescence within two days was reached with doxycycline also in case 2. In comparison to these cases, the course in the third patient was mild, and the patient defervesced spontaneously. CONCLUSION: Mediterranean spotted fever should be considered in the case of unclear fever and rash following a period of residence in the Mediterranean basin during summer time. An eschar may confirm this diagnosis, and early start of therapy may shorten the course of the disease and prevent complications. PMID- 11381637 TI - [Frequent infectious enteritis--2: Therapy]. PMID- 11381638 TI - [Recommendations and prospects of antiretroviral therapy]. PMID- 11381639 TI - [Pathogen resistance--differences in epidemiology]. PMID- 11381640 TI - [Bacterial endocarditis]. PMID- 11381641 TI - [Using neuraminidase inhibitors in prevention and treatment of influenza]. PMID- 11381642 TI - [When the butterfly mosquito meets the dog: leishmaniasis]. PMID- 11381643 TI - [New guidelines of the European Union. Motivation for drug testing in pediatrics]. PMID- 11381644 TI - [Effective in HIV: dual combination with indinavir and ritonavir]. PMID- 11381645 TI - Calibration and other GFCI issues. PMID- 11381646 TI - Ready reminders. PMID- 11381647 TI - A 'not so secret' weapon. PMID- 11381648 TI - Summer breezes--but indoors. PMID- 11381649 TI - Pushing productivity. PMID- 11381650 TI - Estimating cartridge service life. PMID- 11381651 TI - Training safety committee members. PMID- 11381652 TI - An innovative ergo tool. PMID- 11381653 TI - Measuring bioaerosols. PMID- 11381654 TI - Line blurs between professions. PMID- 11381655 TI - Advice on defective sampling data. PMID- 11381656 TI - A sensor of accomplishment. PMID- 11381657 TI - How much intrinsic safety is enough? PMID- 11381658 TI - Seeing is believing. PMID- 11381659 TI - Rethinking the zone of safety. PMID- 11381660 TI - LP creates a safety culture. PMID- 11381661 TI - Preventing gastrointestinal complications of NSAIDs. Risk factors, recent advances, and latest strategies. AB - In the United States, gastrointestinal complications induced by nonsteroidal anti inflammatory drugs (NSAIDs) cause more than 100,000 hospitalizations and an estimated 16,500 deaths annually. Because serious gastrointestinal events can occur without warning, prevention measures must not rely on warning signs alone. This article discusses the epidemiology of NSAID-induced toxic episodes, reviews the risk factors for these occurrences, and offers strategies for minimizing the risk among long-term NSAID users. PMID- 11381662 TI - What patients do--and don't--understand. PMID- 11381663 TI - Clues to an elusive effusion. Postpericardiotomy syndrome. PMID- 11381664 TI - Legionnaires' disease and pneumonia. Beware the temptation to underestimate this "exotic" cause of infection. AB - Legionnaires' disease is an often overlooked but common cause of community acquired pneumonia. The clinical presentation is nonspecific, although fever higher than 39 degrees C (102.2 degrees F), gastrointestinal symptoms, and hyponatremia should raise the index of suspicion. In this article, Drs Akbas and Yu describe specialized laboratory tests needed for definitive diagnosis and discuss therapeutic options. PMID- 11381666 TI - Lower leg edema and lichenification. Elephantiasis nostras verrucosa. PMID- 11381665 TI - Benign vulvar disorders. Identifying features, practical management of nonneoplastic conditions and tumors. AB - Benign vulvar disorders are infrequent, but symptoms such as pruritus, pain, burning, and irritation or the finding of a mass or growth can cause significant patient concern. In this article, Drs Larrabee and Kylander differentiate common nonneoplastic disorders and benign tumors and offer practical management options. PMID- 11381667 TI - Who should receive hepatitis B vaccinations? PMID- 11381668 TI - "Wet" label can be dangerously misleading. PMID- 11381669 TI - Reducing clip-on clutter. The pros and cons of multipurpose communications tools. PMID- 11381670 TI - Inhaled corticosteroids as first-line therapy for asthma. Why they work--and what the guidelines and evidence suggest. AB - Despite a plethora of clinical guidelines and evidence outlining the efficacy of inhaled corticosteroids, these medications continue to be underused in the treatment of asthma. This article reviews the justification for prescribing inhaled steroids as first-line therapy, discusses indications for their use, and compares potencies of the various products now available. In addition, the authors address the adverse effects commonly associated with these agents and provide evidence supporting early intervention with inhaled steroids in the treatment of persistent asthma. PMID- 11381671 TI - How to manage difficult asthma cases. An action plan for physicians and patients. AB - Patients with difficult-to-manage asthma represent one of the greatest challenges in primary care practice. Because these patients are at risk for life-threatening attacks, close monitoring is essential to ensure compliance and to control attacks or medication side effects. In this article, Dr Hunt discusses the many aspects of management, including assessment, identification of triggers, treatment planning, and patient and family education. An "action plan" that outlines the management program is described. PMID- 11381672 TI - Which diagnostic tests for common allergies? Where to start when you face an allergy puzzle. AB - The picture is all too common. In fact, you probably see it in about 1 of every 10 patients who come through your door. The runny nose, scratchy throat, itchy eyes, recurrent sneezing, and annoying cough all point to an allergy. But where do you start and how far do you go in your search for the cause? In this article, Dr Volcheck describes the most commonly used tests for detecting sensitivity to specific IgE allergens and discusses the clinical usefulness and practicality of the various diagnostic procedures. PMID- 11381673 TI - Immunotherapy for inhalant allergies. Guidelines for why, when, and how to use this treatment. AB - Great-grandma was probably on the right track when she said, "A little dirt won't hurt you. It builds immunity." In fact, some evidence suggests that our "hygienic" environment may be increasing our vulnerabilities to some maladies, especially allergic disorders. Although the principles of immunotherapy have been known for nearly a century, guidelines for its use are still evolving. In this article, Dr Portnoy discusses the role of immunotherapy in patients with symptoms triggered by airborne allergens. PMID- 11381674 TI - Comparability of cause of death between ICD-9 and ICD-10: preliminary estimates. AB - OBJECTIVES: This report presents preliminary results describing the effects of implementing the Tenth Revision of the International Classification of Diseases (ICD-10) on mortality statistics for selected causes of death effective with deaths occurring in the United States in 1999. The report also describes major features of the Tenth Revision (ICD-10), including changes from the Ninth Revision (ICD-9) in classification and rules for selecting underlying causes of death. Application of comparability ratios is also discussed. METHODS: The report is based on cause-of-death information from a large sample of 1996 death certificates filed in the 50 States and the District of Columbia. Cause-of-death information in the sample includes underlying cause of death classified by both ICD-9 and ICD-10. Because the data file on which comparability information is derived is incomplete, results are preliminary. RESULTS: Preliminary comparability ratios by cause of death presented in this report indicate the extent of discontinuities in cause-of-death trends from 1998 through 1999 resulting from implementing ICD-10. For some leading causes (e.g., Septicemia, Influenza and pneumonia, Alzheimer's disease, and Nephritis, nephrotic syndrome and nephrosis), the discontinuity in trend is substantial. The ranking of leading causes of death is also substantially affected for some causes of death. CONCLUSIONS: Results of this study, although preliminary, are essential to analyzing trends in mortality between ICD-9 and ICD-10. In particular, the results provide a means for interpreting changes between 1998, which is the last year in which ICD-9 was used, and 1999, the year in which ICD-10 was implemented for mortality in the United States. PMID- 11381675 TI - Effect of a platelet-activating factor receptor antagonist, E5880, on cerebral vasospasm after aneurysmal subarachnoid hemorrhage--open clinical trial to investigate efficacy and safety. AB - The efficacy and safety of a new platelet-activating factor receptor antagonist, E5880, were investigated for preventing cerebral vasospasm after subarachnoid hemorrhage (SAH) in 71 patients with SAH who underwent surgery for ruptured aneurysms within 3 days. Intravenous E5880 administration (300 micrograms or 1200 micrograms twice daily) was begun within 4 days and continued for 14 days. The incidence of symptomatic vasospasm, low-density area on computed tomography, and angiographic vasospasm was lower than in placebo groups in previous studies. Clinical outcome was favorable compared with previous studies. No clinically important adverse events were observed. These results suggest that E5880 is safe and effective in the treatment of patients with cerebral vasospasm due to SAH. PMID- 11381676 TI - Vasodilatory effect of basic fibroblast growth factor in isolated rat cerebral arterioles: mechanisms involving nitric oxide and membrane hyperpolarization. AB - Basic fibroblast growth factor (bFGF), a potent mitogen, acutely dilates cerebral blood vessels and may be effective in reducing cerebral infarction. However, the vasodilatory mechanism, which may involve nitric oxide (NO), is not completely understood. This study investigated whether membrane hyperpolarization is also involved in this mechanism. Membrane potential (MP) of smooth muscle cells and vessel diameter of isolated intracerebral arterioles were simultaneously measured following extraluminal application of bFGF in rats. The involvement of NO and adenosine triphosphate-sensitive potassium (KATP) channels in bFGF-induced vasodilation and membrane hyperpolarization was evaluated using specific inhibitors, NG-monomethyl-L-arginine (L-NMMA, 10(-4) M) and glibenclamide (GB, 10(-5) M), respectively. The resting MP was recorded at a mean value of -31.9 +/- 4.5 mV. bFGF (1 to 1000 ng/ml) produced significant vasodilation and hyperpolarization. Treatment with L-NMMA caused vasoconstriction and significantly attenuated bFGF-induced vasodilation without affecting membrane hyperpolarization. In the presence of GB, the membrane potential was significantly depolarized but the vessel diameter was only marginally reduced, so bFGF-induced membrane hyperpolarization was inhibited while arteriolar dilation was attenuated. These results suggest that bFGF-induced vasodilation is mediated by a mechanism involving both NO and membrane hyperpolarization, and that membrane hyperpolarization is caused by the activation of KATP channels. PMID- 11381677 TI - Expression of nuclear factor-kappa B, tumor necrosis factor receptor type 1, and c-Myc in human astrocytomas. AB - Tumor necrosis factor receptor type 1 (TNFR1) and c-Myc are important in signal transduction in tumor necrosis factor-alpha (TNF-alpha)-induced cytotoxicity, whereas activation of nuclear factor-kappa B (NF-kappa B) protects against TNF alpha-induced apoptosis. This study investigated the expression of NF-kappa B, TNFR1, and c-Myc in human astrocytoma tissues by reverse transcriptase-polymerase chain reaction (PCR) and immunohistochemical analysis. TNFR1 messenger ribonucleic acid (mRNA) and c-Myc mRNA were frequently expressed in malignant astrocytomas, especially in glioblastomas, compared with low-grade astrocytomas by PCR analysis. TNFR1 and c-Myc mRNAs were barely detectable in normal brain tissues. NF-kappa B p50 and p65 subunit mRNAs were detected in various grades of astrocytomas, with frequent expression in malignant astrocytomas. The presence of activated NF-kappa B was confirmed by nuclear localization in neoplastic astrocytes as determined by immunohistochemistry. Both p50 and p65 subunits were inhomogeneously expressed in neoplastic astrocytes of glioblastoma, but only in a few scattered tumor cells in low-grade astrocytoma, and almost undetectable in normal brain tissues. These results indicate that TNFR1 and c-Myc are overexpressed in malignant astrocytomas, and this may increase the cellular sensitivity to the cytotoxic action of TNF-alpha. NF-kappa B p50 and p65 were simultaneously induced and activated in malignant astrocytomas. Our results suggest that the constitutive activation of NF-kappa B subunits in malignant astrocytoma, especially in glioblastoma, could be associated with the resistance to TNF-alpha immunotherapy, and indicates new therapeutic strategies for malignant astrocytomas. PMID- 11381678 TI - Chronic subdural hematoma with vasogenic edema in the cerebral hemisphere--case report. AB - An 80-year-old male with a history of hypertension presented with chronic subdural hematoma manifesting as progressive consciousness disturbance and left hemiparesis. T1-weighted and fluid attenuation inversion recovery (FLAIR) magnetic resonance imaging showed a fresh hematoma in the right subdural space with a midline shift of 15 mm. FLAIR and diffusion-weighted imaging showed a hyperintense area in the right paraventricular white matter compressed by the hematoma. Apparent diffusion coefficients (ADCs) corresponding to the hyperintense area in the central area of the affected cerebral hemisphere on FLAIR images were measured before and one month after the operation. The motion probing gradient was applied in the right-left direction to the body axis. Since the central area in the cerebrum includes nerve fibers perpendicular to the direction of the gradient, the measured ADC appeared to be anisotropic. Preoperative ADC in the right paraventricular white matter was anisotropic and greater than in age-matched normal subjects, so the edema was identified as the vasogenic type. The edema in the right paraventricular white matter resolved promptly with improvement of the midline shift and normalization of the ADC. PMID- 11381679 TI - Tentorial vascularization in solid hemangioblastoma--case report. AB - A 40-year-old female was admitted to the hospital with complaints of headache worsening gradually over a 1-month duration. Her past history included surgery to treat a left cerebellar cystic lesion 3 years before, and an untreated small solid right supracerebellar lesion of 1 cm diameter. On admission, magnetic resonance imaging showed that the right cerebellar lesion had grown to approximately 4 cm diameter abutting the tentorium and causing obstructive hydrocephalus. She also had two more small lesions, a right supratentorial solid lesion with cystic component near the splenium and an intramedullary cystic lesion at the C-2 level. Right suboccipital craniectomy was done. The vascular attachments between the superior aspect of the tumor and the tentorium were coagulated and the tumor was totally removed. C1-2 laminectomy was also performed to drain the intramedullary cyst. The patient deteriorated and lost consciousness with respiratory arrest 6 hours postoperatively and was reoperated for intracerebellar hematoma due to oozing from the tentorial vessels. Histological investigation revealed hemangioblastoma. Dural tentorial vascular attachments in solid hemangioblastomas located subjacent to the tentorium may cause early postoperative complications of hematoma at the site of vascular attachment following the resection. Computed tomography study in the early postoperative period is helpful to identify this problem. PMID- 11381680 TI - Primary Ewing's sarcoma of the occipital bone--case report. AB - A 17-year-old female presented with a very rare case of primary Ewing's sarcoma of the skull involving the occipitotemporal region. Systemic examination found no evidence of metastasis. The tumor was surgically removed, and the patient underwent radiotherapy and chemotherapy. Fourteen months after surgery there has been no recurrence of the tumor. Cranial primary Ewing's tumor has a good prognosis after radical surgery and adjuvant therapy. PMID- 11381681 TI - Mazindol administration improved hyperphagia after surgery for craniopharyngioma- case report. AB - A 54-year-old man presented with visual disturbance and polydipsia. Magnetic resonance imaging disclosed a cystic mass which extended from the intrasellar to the suprasellar region. Bifrontal craniotomy was performed and the tumor was totally removed. Histological findings confirmed the diagnosis of craniopharyngioma. Postoperatively, the patient suffered from transient disorientation. About one month after the operation the patient manifested hyperphagia and he gained 15 kg in one month. Mazindol, a non-amphetaminergic anorectic agent, was administered for 3 weeks. His appetite normalized and his weight fell and stabilized even after mazindol administration was ceased. PMID- 11381682 TI - Squamous cell carcinoma arising in an intracranial dermoid cyst--case report. AB - A 47-year-old male presented with headache. Magnetic resonance imaging revealed an enhanced mass lesion in the cerebellar vermis and left cerebellar hemisphere and in the cisterna magna. Gross total removal of the tumor was performed. Histological examination demonstrated squamous cell carcinoma in the dermoid cyst. The patient subsequently received localized radiation therapy of total 55 Gy. He has been without tumor recurrence for 6 years since the surgery. We recommend local radiation therapy over 50 Gy following surgery. PMID- 11381683 TI - Transverse sinus-tentorium splitting approach for pineal region tumors--case report. AB - A 15-year-old boy with a large pineal region mass was admitted to our institute. The tentative diagnosis was mixed germ cell tumor. Tumor resection was carried out via a transverse sinus-tentorium splitting approach. The tumor tissue was completely resected, and no operative complication other than transient vertical gaze paresis was noted. The histological diagnosis was mixed germ cell tumor composed of mature and immature teratoma, germinoma, and embryonal carcinoma. After three courses of chemotherapy, the patient underwent external irradiation. He remained asymptomatic with no signs of recurrence 42 months after the surgery. The combination of the infratentorial supracerebellar approach and the occipital transtentorial approach provides excellent views and work space above and below the tentorial notch. Transverse sinus section is not mandatory for this approach, but sectioning of the unilateral transverse sinus and the tentorium along the rectal sinus allows retraction of the falx and the underlying brain to the opposite side. Thus, a much wider horizontal and vertical projection is obtained. This approach enables safer and more extensive tumor removal for large pineal region tumors. PMID- 11381684 TI - Transoral transclival approach for intradural lesions using a protective bone baffle to block cerebrospinal fluid pulse energy--two case reports. AB - The transoral transclival approach for the treatment of intradural lesions of the clivus is often associated with serious complications such as cerebrospinal fluid (CSF) leakage and meningitis. CSF pulse energy may be the most significant factor in CSF leakage and meningitis, but a bone baffle can block such CSF pulse energy. A 64-year-old female presented with sudden onset of severe headache. She had subarachnoidal hemorrhage due to a rupture of the vertebral-posterior inferior cerebellar artery aneurysm. A 66-year-old female complaining of occipitalgia and numbness of the extremities had a foramen magnum meningioma. Both patients were treated via the transoral transclival route with a protective bone baffle, obtained from the iliac bone, securely fixed in the bone window to protect the repaired dura from injury by CSF pulse energy. Neither patient showed CSF leakage or meningitis, and the period of continuous lumbar CSF drainage was only 7 days. The transoral transclival approach with a bone baffle is still very effective in selected cases. PMID- 11381686 TI - AFM observation of surface acoustic waves emitted from single symmetric SAW transducers. AB - We report the first experimental observation of surface acoustic waves (SAWs) launched from a single symmetric SAW transducer, employing scanning acoustic force microscopy (SAFM). SAFM is a simple technique for the imaging of complex interdigital transducer (IDT) radiation patterns with nanometer lateral resolution. We demonstrate submicron lateral resolution and high sensitivity by investigating a single excitation element on a weakly coupling substrate (GaAs), visualizing the launched wave and second-order effects. PMID- 11381685 TI - Micromachining of a piezocomposite transducer using a copper vapor laser. AB - A 1-3 piezocomposite transducer with front face dimensions of 2 x 2 mm has been micromachined using a copper vapor laser. The device consists of PZT5A piezoceramic pillars with a 65-micron pitch suspended in a low viscosity thermosetting polymer. The kerf width is 13 microns, and the transducer thickness is 170 microns, making the device suitable for ultrasonic reception at frequencies close to 10 MHz. PMID- 11381687 TI - A new ultrasound contrast imaging approach based on the combination of multiple imaging pulses and a separate release burst. AB - A new ultrasound contrast imaging technique is described that optimally employs the rupture of the contrast agent. It is based on a combination of multiple high frequency, broadband, imaging pulses and a separate release burst. The imaging pulses are used to survey the target before and after the rupture and release of free gas bubbles. In this way, both processes (imaging and release) can be optimized separately. The presence of the contrast agent is simply detected by correlating or subtracting the signal responses of the imaging pulses. Because the time delay between the imaging pulses can be very short, the subtraction is less affected by tissue motion and can be done in real time. In vitro measurements showed that by using a release burst, the detection sensitivity increased 12 to 43 dB for different types of contrast agents. In the presence of a moving phantom, the increase in sensitivity was 22 dB. This new method is very sensitive for contrast agent detection in fundamental imaging mode and, therefore, non-linear propagation effects do not limit the maximum obtainable agent-to-tissue ratio. However, because of the inherent destruction of the contrast agent, it has to operate in an intermittent way. Through experiments, we have demonstrated the potential of the method to achieve simultaneous high sensitivity for contrast detection, i.e., high agent-to-tissue ratio, and high spatial resolution performance for different types of contrast agents. PMID- 11381688 TI - Surface acoustic wave impedance element ISM duplexer: modeling and optical analysis. AB - Surface acoustic wave (SAW) impedance element antenna duplexers provide compact, high performance, front-end components apt for industrial fabrication. We describe investigations on the design and modeling of a compact ISM antenna duplexer fabricated on a 36 degrees YX-cut LiTaO3 substrate based on SAW impedance elements. In particular, we have performed 3-D modeling of the inductive and capacitive electromagnetic couplings caused by the package parasitics for the duplexer. The use of a 1:3 IDT structure for the reduction of the passband width is discussed. The frequency response of the duplexer is predicted with the help of circuit simulation; the modeling is refined by optimization of the model parameters to improve the fit between the measured and simulated responses. We also report scanning optical imaging of the acoustic field within the resonator structures with the help of laser interferometry; this provides insight into the loss mechanisms beyond that attainable in mere electric measurements. PMID- 11381689 TI - The adiabatic anti-jitter circuit. AB - The anti-jitter circuit (AJC) [1], [2]-[4] is able to reduce phase noise and spurious content of any frequency source at sideband frequencies above a defined cut-off frequency. By contrast, a phase-locked loop (PLL) can only reduce the intrinsic phase noise of its output oscillator closer to carrier lower than a defined cut-off frequency. The AJC has no output oscillator, but its phase noise performance can be assessed as if it had [5]. This paper reports the invention of the adiabatic AJC (AAJC) [6]-[8], giving the AJC improved power consumption, frequency range, and maximum frequency of operation. The term "adiabatic" is adopted to indicate that the core part of the new circuit does not require a power supply. It takes power from the input source directly to create the sawtooth waveform that has considerably reduced time jitter on the longer of its two ramp waveforms. Discrete models of the AJC are now operational at 30 MHz, which is twice the 15-MHz operation previously reported. The cut-off frequency of suppression has been maintained at a few kiloHertz. Noise analysis now shows performance comparable with an LC oscillator is possible. SPICE simulations show potential operation up to 5 GHz. The AAJC is also cascadable up to the intrinsic (shot) noise limit. Shot noise can be reduced by feedback. PMID- 11381690 TI - Thermostatic and dynamic performance of an ultrasonic density probe. AB - The thermally static and dynamic performance of an ultrasonic density probe for liquids is investigated in the density range of 750 to 1300 kg/m3 at temperature ranging from 0 to 40 degrees C. The single transducer probe uses a pulse echo technique to obtain the characteristic acoustic impedance of the liquid and, subsequently, the speed of sound through the liquid to obtain the density of the liquid. Variations in the initial sound amplitude are addressed by the design of a layered two material probe. It is shown that it is possible to obtain an accuracy of 0.4% in the experiments carried out. For changing temperature, the probe exhibits large errors because of problems in estimating the temperatures in certain regions of the probe. PMID- 11381691 TI - Travelling wave ultrasonic motor using the B08 flexural mode of a circular membrane. AB - This paper describes the design, construction, and performance of a piezoelectric motor that uses the travelling B08 mode of an 80-mm diameter circular membrane to drive a rotor by frictional contact. The motor is of a thin planar design, giving high torque of up to 0.33 Nm at low speed and has been developed as a design that can be made with lithographic techniques for miniaturization. Investigations of the free stator with a vibration pattern imager and impedance analyzer gave the resonance frequency, mode, and electromechanical coupling of the stator. Motor speed as a function of frequency for a constant voltage and performance charts of speed, output power, and efficiency against torque are presented for a particular input voltage and rotor pre-load. The effects of two different lead zirconate titanate (PZT) ring dimensions have been investigated. Excitation of the B09 mode has been observed, incommensurate with the piezoelectric excitation of the stator. This is discussed with relation to edge-clamping of the stator. Single standing wave motoring was observed, believed to arise from asymmetry of the stator and its perturbation of the B08 resonance mode. Sources of power loss, including frame vibration and friction interface slip, are considered and discussed. PMID- 11381692 TI - Near point-source longitudinal and transverse mode ultrasonic arrays for material characterization. AB - Specially constructed near point-source ultrasonic transducers (0.75 MHz) were designed to preferentially stimulate and receive the one longitudinal (P) and two transverse (S) propagation modes. Arrays of these transducers were placed on a rectangular prism of common sodalime glass, which served as an ideal homogeneous, isotropic medium, to evaluate the uncertainty of a newly developed phase velocity measurement method. Through the use of the Radon transform, the data were transformed from the offset-time (x-t) domain to the intercept time--horizontal slowness (tau-p) domain. From the shape of the curves in the tau-p domain, the phase velocity of the propagating waves may be determined for a range of directions. The phase velocities determined using this method were accurate for incidence angles up to 76 degrees, 64 degrees, and 77 degrees for the P, SV, and SH wave modes, respectively. Phase velocities of 5724 +/- 64, 3411 +/- 30, and 3467 +/- 15 m/s were determined for the P-wave, SV-wave, and SH-wave modes, respectively. This agrees with the direct transmission P-wave and S-wave velocities of 5690 +/- 60 and 3440 +/- 26 m/s, respectively, to better than 1%. PMID- 11381693 TI - A standing wave-type noncontact linear ultrasonic motor. AB - In this study, a novel standing wave-type noncontact linear ultrasonic motor is proposed and analyzed. This linear ultrasonic motor uses a properly controlled ultrasonic standing wave to levitate and drive a slider. A prototype of the motor was constructed by using a wedge-shaped aluminum stator, which was placed horizontally and driven by a multilayer PZT vibrator. The levitation and motion of the slider were observed. Assuming that the driving force was generated by the turbulent acoustic streaming in the boundary air layer next to the bottom surface of the slider, a theoretical model was developed. The calculated characteristics of this motor were found to agree quite well with the experimental results. Based on the experimental and theoretical results, guidelines for increasing the displacement and speed of the slider were obtained. It was found that increasing the stator vibration displacement, or decreasing the gradient of the stator vibration velocity and the weight per unit area of the slider, led to an increase of the slider displacement. It was also found that increasing the amplitude and gradient of the stator vibration velocity, or decreasing the weight per unit area of the slider and the driving frequency, gave rise to an increase of the slider speed. There exists an optimum roughness of the bottom surface of the slider at which the slider speed has a maximum. PMID- 11381694 TI - Doppler ultrasound signal denoising based on wavelet frames. AB - A novel approach was proposed to denoise the Doppler ultrasound signal. Using this method, wavelet coefficients of the Doppler signal at multiple scales were first obtained using the discrete wavelet frame analysis. Then, a soft thresholding-based denoising algorithm was employed to deal with these coefficients to get the denoised signal. In the simulation experiments, the SNR improvements and the maximum frequency estimation precision were studied for the denoised signal. From the simulation and clinical studies, it was concluded that the performance of this discrete wavelet frame (DWF) approach is higher than that of the standard (critically sampled) wavelet transform (DWT) for the Doppler ultrasound signal denoising. PMID- 11381695 TI - The use of air-coupled ultrasound to test paper. AB - Capacitance transducers containing a thin polymer membrane have been used to transmit ultrasonic signals with frequencies in excess of 1 MHz through various paper products such as paper and cardboard. At normal incidence, a resonance was visible in thicker samples, the frequency of which could be correlated to parameters such as the thickness of the paper sample and the moisture content. It has also been demonstrated that images can be obtained of changes in structure across paper and card samples. PMID- 11381696 TI - Harmonic leakage and image quality degradation in tissue harmonic imaging. AB - Image quality degradation caused by harmonic leakage was studied for finite amplitude distortion-based harmonic imaging. Various sources of harmonic leakage, including transmit waveform, signal bandwidth, and system nonlinearity, were investigated using both simulations and hydrophone measurements. Effects of harmonic leakage in the presence of sound velocity inhomogeneities were also considered. Results indicated that sidelobe levels of the harmonic beam pattern were directly affected by harmonic leakage when the harmonic signal was obtained by filtering out the fundamental signal. Because sidelobe levels also increase with the bandwidth of the transmitted signal, a trade-off exists between axial resolution and contrast resolution. It is concluded that accurate control of the frequency content of the waveform prior to propagation is necessary to optimize imaging performance of tissue harmonic imaging. The filtering technique was also compared with the pulse inversion technique. It was shown that the pulse inversion technique effectively suppresses harmonic leakage at the cost of imaging frame rate and potential motion artifacts. PMID- 11381697 TI - A thin film phantom for blood flow simulation and Doppler test. AB - The thin film phantom is a new type of ultrasound resolution test object. It consists of a thin planar substrate that is acoustically matched to the surrounding media. Precisely located scatterers on the surface of the substrate generate echo signals. The patterning of scatterers on the substrate allows echogenicity to be controlled as a function of position, which enables the production of a test object with highly reproducible and controllable scattering characteristics. We show that by vibrating the substrate in a suitable manner, an echo signal may be generated that simulates bi-directional flow. We demonstrate that a vibration of low amplitude at frequency f0 produces a Doppler spectral signal at f0 and -f0, within the limits of aliasing. Furthermore, by driving the film with a bandlimited noise signal, we illustrate how a velocity distribution may be simulated. A time-varying flow velocity may be simulated by varying the noise bandwidth with time. Finally, using this technique, we demonstrate a system that simulates an arterial flow pattern, including its characteristic velocity distribution in forward and reverse directions simultaneously. PMID- 11381698 TI - Novel interferometric frequency discriminators for low noise microwave applications. AB - New configurations of interferometric frequency discriminators (FD) for frequency stabilization of microwave oscillators are examined. The new FDs are arranged in single directional (SD) (patented), bi-directional (BD) (patent pending), and dual reflection (DR) (patent pending) configurations. In the SD configuration, the signals reflected off and transmitted through the resonator separately pass through different arms of the interferometer. In the BD configuration, microwaves pass in both directions through each arm of the interferometer. In the DR configuration, microwaves are reflected from the resonator as well as the compensating arm. The FD sensitivity is compared with that for the conventional interferometric FD and found to be 6 dB greater in the BD configuration. Because no circulator is required within the interferometer in either the BD or the DR FD, the discriminator's phase noise floor is not limited by the circulator contribution. PMID- 11381699 TI - Characterization of one-dimensional capacitive micromachined ultrasonic immersion transducer arrays. AB - In this paper, we report on the characterization of 1-D arrays of capacitive micromachined ultrasonic transducers (cMUT). A 275- x 5600-micron 1-D CMUT array element is experimentally characterized, and the results are found to be in agreement with theoretical predictions. As a receiver, the transducer has a 0.28 fm/square root of Hz displacement sensitivity, and, as a transmitter, it produces 5 kPa/V of output pressure at the transducer surface at 3 MHz with a DC bias of 35 V. The transducer has more than 100% fractional bandwidth around 3 MHz, which makes it suitable for ultrasound imaging. The radiation pattern of isolated single elements, as well as those of array elements are measured, and two major sources of acoustical cross talk are identified. A weakly dispersive non-leaky interface wave (Stoneley wave) is observed to be propagating at the silicon substrate-fluid interface at a speed close to the speed of sound in the fluid. This wave causes internal reflections, spurious resonance, and radiation from the edges of the silicon substrate. The large lateral component of the particle velocity generated by the membranes at the edge of the cMUT array elements is found to be the source of this interface wave. Lowest order Lamb waves in the silicon substrate are also found to contribute to the cross talk between elements. These waves are excited at the edges of individual vibrating membranes, where they are anchored to the substrate, and result in a narrowing of the beam profile of the array elements. Several methods, such as trench isolation and wafer thinning, are proposed and implemented to modify the acoustical cross coupling between array elements. PMID- 11381701 TI - Scattering analysis of two-port SAW resonators. AB - Linear equations derived from the scattering matrix approach to the two-port resonator were solved, and analytical expressions for the normalized SAW amplitudes were obtained. Asynchronous and synchronous resonators were analyzed numerically. It was shown that the output of the two-port resonator is a sum of two signals. In the case of the asynchronous resonator, these signals are in phase at a resonance frequency; for the synchronous resonator, they are in phase quadrature, which causes the higher insertion loss of the synchronous resonator. PMID- 11381700 TI - Implementing guided wave mode control by use of a phased transducer array. AB - A multi-channel time-delay system has been built and applied to a transducer array for implementing guided wave mode control. The time-delay system has a capability of sending high energy controllable tone-burst signals from eight independent channels with arbitrary time delays from 0 to 30 microseconds with resolution of 0.025 microsecond. Software time delays are also provided for summing up received signals of each channel. Theoretical discussions indicate the impact of the time delay capability on the bandwidth and sensitivity improvement of a transducer array for guided wave generation. Determination of both physical and software time delay values is based on a knowledge of dispersion curves and element spacing. Based on reference signals, a non-knowledge-based automatic time delay searching algorithm was introduced for guided wave mode selection. Experiments were conducted with a phased comb transducer array mounted on a carbon steel pipe. The experimental results show that signal to noise ratio has been greatly improved by use of the time-delay system. Some other benefits of the phased array, including unidirection generation and mode control flexibility, are discussed. PMID- 11381702 TI - Bulk acoustic wave sensors for sensing measurand-induced electrical property changes in solutions. AB - A variety of quartz thickness shear mode (TSM) resonant sensors with different electrode configurations have been designed, fabricated, and tested in liquids for probing liquid electrical property changes. The resonant frequency of the sensors was found to have more than an order of magnitude increase in sensitivity over the standard AT-cut quartz resonant sensor. The increase in sensitivity is due to the ability of the sensors to detect changes in the electrical properties in the liquid. PMID- 11381703 TI - Finite element simulation of nonlinear wave propagation in thermoviscous fluids including dissipation. AB - A recently developed finite element method (FEM) for the numerical simulation of nonlinear sound wave propagation in thermoviscous fluids is presented. Based on the nonlinear wave equation as derived by Kuznetsov, typical effects associated with nonlinear acoustics, such as generation of higher harmonics and dissipation resulting from the propagation of a finite amplitude wave through a thermoviscous medium, are covered. An efficient time-stepping algorithm based on a modification of the standard Newmark method is used for solving the non-linear semidiscrete equation system. The method is verified by comparison with the well-known Fubini and Fay solutions for plane wave problems, where good agreement is found. As a practical application, a high intensity focused ultrasound (HIFU) source is considered. Impedance simulations of the piezoelectric transducer and the complete HIFU source loaded with air and water are performed and compared with measured data. Measurements of radiated low and high amplitude pressure pulses are compared with corresponding simulation results. The obtained good agreement demonstrates validity and applicability of the nonlinear FEM. PMID- 11381704 TI - Model-based estimation of ultrasonic echoes. Part I: analysis and algorithms. AB - The patterns of ultrasonic backscattered echoes represent valuable information pertaining to the geometric shape, size, and orientation of the reflectors as well as the microstructure of the propagation path. Accurate estimation of the ultrasonic echo pattern is essential in determining the object/propagation path properties. In this study, we model ultrasonic backscattered echoes in terms of superimposed Gaussian echoes corrupted by noise. Each Gaussian echo in the model is a nonlinear function of a set of parameters: echo bandwidth, arrival time, center frequency, amplitude, and phase. These parameters are sensitive to the echo shape and can be linked to the physical properties of reflectors and frequency characteristics of the propagation path. We address the estimation of these parameters using the maximum likelihood estimation (MLE) principle, assuming that all of the parameters describing the shape of the echo are unknown but deterministic. In cases for which noise is characterized as white Gaussian, the MLE problem simplifies to a least squares (LS) estimation problem. The iterative LS optimization algorithms when applied to superimposed echoes suffer from the problem of convergence and exponential growth in computation as the number of echoes increases. In this investigation, we have developed expectation maximization (EM)-based algorithms to estimate ultrasonic signals in terms of Gaussian echoes. The EM algorithms translate the complicated superimposed echoes estimation into isolated echo estimations, providing computational versatility. The algorithm outperforms the LS methods in terms of independence to the initial guess and convergence to the optimal solution, and it resolves closely spaced overlapping echoes. PMID- 11381705 TI - Model-based estimation of ultrasonic echoes. Part II: Nondestructive evaluation applications. AB - Accurate estimation of the ultrasonic echo pattern leading to the physical property of the object is desirable for ultrasonic NDE (nondestructive evaluation) applications. In Part I of this study, we have presented a generalized parametric ultrasonic echo model, composed of a number of Gaussian echoes corrupted by noise, and algorithms for accurately estimating the parameters. In Part II of this study, we explore the merits of this model-based estimation method in ultrasonic applications. This method produces high resolution and accurate estimates for ultrasonic echo parameters, i.e., time of flight (TOF) amplitude, center frequency, bandwidth, and phase. Furthermore, it offers a solution to the deconvolution problem for restoration of the target response, i.e., ultrasonic reflection and transmission properties of materials, from the backscattered echoes. The model-based estimation method makes deconvolution possible in the presence of significant noise. It can also restore closely spaced overlapping echoes beyond the resolution of the measuring system. These properties of the estimation method are investigated in various ultrasonic applications such as transducer pulse-echo wavelet estimation, subsample time delay estimation, and thickness sizing of thin layers. PMID- 11381706 TI - Analysis of the rutile-ring method of frequency-temperature compensating a high-Q whispering gallery sapphire resonator. AB - The rutile-ring method of dielectrically frequency-temperature compensating a high-Q whispering gallery (WG) sapphire resonator is presented. Two and three dimensional finite element (FE) analysis has been implemented to design and analyze the performance of such resonators, with excellent agreement between theory and experiment. A high-Q factor of 30 million at 13 GHz and compensation temperature of 56 K was obtained. It is shown the frequency-temperature compensation can occur either because the rutile adds a small perturbation to the sapphire resonator or because of a mode interaction with a resonant mode in the rutile. The characteristics of both of these methods are described, and it is shown that for high frequency stability, it is best to compensate perturbatively. PMID- 11381707 TI - Compact, high-Q, zero temperature coefficient, TE011 sapphire-rutile microwave distributed Bragg reflector resonators. AB - Some novel new resonator designs based on the distributed Bragg reflector are presented. The resonators implement a TE011 resonance in a cylindrical sapphire dielectric, which is confined by the addition of rutile and sapphire dielectric reflectors at the end faces. Finite element calculations are utilized to optimize the dimensions to obtain the highest Q-factors and zero frequency-temperature coefficient for a resonator operating near 0 degree C. We show that a Q-factor of 70,000 and 65,000 can be achieved with and without the condition of zero frequency-temperature coefficients, respectively. PMID- 11381708 TI - Measuring Lamb wave dispersion curves of a bi-layered plate and its application on material characterization of coating. AB - This paper investigates the Lamb wave dispersion curves of a bi-layered plate and evaluates the feasibility of using the dispersion data to characterize the coating's material properties. The measurement of dispersion curves is based on a focusing PVDF transducer operating in a pulse/echo mode. An image displaying technique is used to determine the dispersion relation of Lamb waves from the measured data. Multiple dispersion curves of Lamb waves are accurately determined over a wide frequency range (4 to 20 MHz). Lamb wave dispersion curves for thin metal sheets electro-deposited with nickel coatings are measured. The elastic constants of the nickel coating are determined by comparing the experimental dispersion data with the theoretical ones calculated numerically. Potential applications of this measurement method are addressed. PMID- 11381709 TI - Model and measurements of the electrical input impedance of a plate-liquid-plate acoustic resonator. AB - A plate-liquid-plate (PLP) acoustic resonator is used to measure the acoustic velocity of liquids. An analytical model of the electrical input impedance of the resonator is proposed. Theoretical results are compared with experimental measurements using reference liquids. Accuracy of the sound velocity calculation is found to be better than 1%. Finally, factors limiting the achievable accuracy are discussed. PMID- 11381710 TI - Discussion of one-D piezoelectric transducer models with loss. AB - Two, 1-dimensional piezoelectric transducer models are presented that use complex numbers to represent mechanical and dielectric loss. Exact numerical agreement was achieved by avoiding a number of small errors in the literature. A correction to Kino's simplified Mason parallel equivalent circuit is also given. Efficiency is discussed. PMID- 11381711 TI - Reduction of the cold collisions frequency shift in a multiple velocity fountain: a new proposal. AB - In this paper, we propose a new scheme for operating an atomic fountain frequency standard. A sequence of time-spaced balls of atoms, launched over non-overlapping trajectories, makes possible a reduction of the atomic density and consequently the cold collision frequency shift by one order of magnitude without significant reduction of the useful signal and of the overall stability of the clock. The ultimate accuracy of a Cs fountain could be improved below the 10(-15) level currently achieved in operating clocks. PMID- 11381712 TI - Angelo S. Agro, MD. President of the Medical Society of New Jersey. PMID- 11381713 TI - Getting results. Physician advocacy in the age of managed care. PMID- 11381714 TI - Skin cancer update. Treatment alternatives for basal-cell and squamous-cell carcinoma. PMID- 11381715 TI - Community health assessments. Determining their impact. PMID- 11381716 TI - Physician compensation. The benefits of a hybrid system. PMID- 11381717 TI - Physicians gain subpoena power in hospital proceedings. PMID- 11381718 TI - Commercial health plan participation in Medicaid managed care: an examination of six markets. AB - This study examines six local health care markets to gain a better understanding of the factors associated with the decision by commercial plans to participate in Medicaid managed care (MMC). Findings suggest that no single factor explained why plans chose to participate in MMC in a particular market. Instead, a combination of factors--generally economic but not always--determined whether a plan participated. While rate adequacy was central, it was not the only factor. Results indicate that it is capitation rates relative to other factors (such as provider costs, administrative costs, enrollment volume, growth opportunities in other markets) that matter rather than simply the level of rates. PMID- 11381719 TI - Tax cuts and cynicism. PMID- 11381720 TI - Health plan choice and information about out-of-pocket costs: an experimental analysis. AB - Many consumers are offered two or more employer-sponsored health insurance plans, and competition among health plans for subscribers is promoted as a mechanism for balancing health care costs and quality. Yet consumers may not receive the information necessary to make informed health plan choices. This study tests the effects on health plan choice of providing supplemental decision-support materials to inform consumers about expected health plan costs. Our main finding is that such information induces consumers to bear more risk, especially those in relatively good health. Thus our results suggest that working-age, privately insured consumers currently may be over-insuring for medical care. PMID- 11381721 TI - Racial disparities in service use among Medicaid beneficiaries after mandatory enrollment in managed care: a difference-in-differences approach. AB - Managed care may improve access to health care to previously underserved populations when providers need plan enrollees. However, capitation and utilization management often give providers the incentive to withhold care. Managed care organizations have yet to demonstrate that racial disparities in treatment are not exacerbated. Using Medicaid eligibility, claims, and managed care encounter data, we examine racial disparities in service use among Medicaid beneficiaries after mandatory enrollment in managed care. We use count data models adjusted for nonrandom selection within difference-in-differences econometric approaches. The results show that mandatory enrollment has disproportionately reduced the relative use of physician and inpatient services among African-American beneficiaries. PMID- 11381722 TI - What accounts for differences in uninsurance rates across communities? AB - Using data from the 1996-97 Community Tracking Study household survey, this study examines variations in uninsurance rates across communities in the United States. Specifically, regression-based decomposition is used to identify factors that account for high rates of uninsurance in some communities. Differences in explained rates between "high uninsurance" and "low uninsurance" communities are the result of differences in the racial/ethnic composition and socioeconomic status of the population (33%), differences in employment characteristics (26%), and state Medicaid eligibility requirements (12.7%). Although higher costs are associated with a higher likelihood that individuals are uninsured, high-cost communities tend to have lower rates of uninsurance as a result of other factors. Despite the large number of identifiable factors included in the analysis, there is still a substantial amount of unexplained regional variation in uninsurance rates. PMID- 11381723 TI - PACE and the Medicare+Choice risk-adjusted payment model. AB - This paper investigates the impact of the Medicare principal inpatient diagnostic cost group (PIP-DCG) payment model on the Program of All-Inclusive Care for the Elderly (PACE). Currently, more than 6,000 Medicare beneficiaries who are nursing home certifiable receive care from PACE, a program poised for expansion under the Balanced Budget Act of 1997. Overall, our analysis suggests that the application of the PIP-DCG model to the PACE program would reduce Medicare payments to PACE, on average, by 38%. The PIP-DCG payment model bases its risk adjustment on inpatient diagnoses and does not capture adequately the risk of caring for a population with functional impairments. PMID- 11381724 TI - Costs in the last year of life in The Netherlands. AB - The costs of health care in the last year of life are a subject of debate and myth. Expensive interventions at the end of life often are blamed for the rapid increase in health care spending, but evidence about the existence of such exceptionally high expenditures at the end of life is rare and faulty. This investigation examines the development and composition of health care costs at the end of life for all age groups in The Netherlands. In contrast with earlier studies, this research analyzes both acute care (cure) and long-term care (care) costs. As an alternative for the frequently used concept of calendar years, we employed the concept of life years for calculating the costs at the end of life. We found that when life approaches its end, health care expenditures indeed rise sharply, especially in the last months. However, when we compared total cure costs in the last year of life to the total cure costs for the entire population, we concluded that the end-of-life share was only about 10%. Results of this study show that interventions to reduce costs in the last year of life will have only a modest impact compared to the total health care budget. PMID- 11381725 TI - ["Perhaps, the night, I cry while I write": strategies for survival of p.v.p. patients]. AB - The history of modern psychiatry has been written by people who have made a career of studying the world of mental health. Especially since the 1960s, many historians, journalists, sociologists and psychiatrists have described and analysed the contradictions of this new science and its applications to specific clienteles. What about psychiatric survivors themselves? How do they view their experience? What is their opinion of the care they have received? Few studies have looked at their written version of psychiatry. This article explores some of the writings signed by survivors of psychiatry to determine how they assess their experience. For this exploratory study, 5 sources are looked at: 1 in Ontario, 2 in Quebec, 1 in New Brunswick and 1 on the World Wide Web. The author stresses the potential of these "voices of experience" and the unique abilities of the victims of psychiatry to describe and analyse the oppression they have been through. PMID- 11381726 TI - [The forgotten actors]. AB - Within the Regional Plans for the Organization of Services, Quebec's mental health reform has tried to redistribute power and influence to mutual aid groups for consumers and their families. However, it has failed to provide them with the financial resources that would allow them to exert that new power. The groups don't have enough personnel to fill the decision-making positions accorded to them under the regional plans while, at the same time, continuing to provide services to consumers. Real partnership requires equality in resources. Social integration being the product of an interaction between consumers and citizens, citizens are the forgotten actors of the interaction. We fail to support them when they are faced with behaviours that they find strange and disturbing. Attitudinal and behavioural changes are interrelated and concomitant; that is, the transformation of citizens' prejudice into acceptance is helped by consumers' efforts to adopt less offensive behaviours. PMID- 11381727 TI - From humane care to prevention. AB - Dr. Leonard R. Denton's professional career has spanned the fields of education, social work, program evaluation, research, and clinical psychology, in both institutional and community mental health settings. He is an advocate for community mental health and prevention programs aimed at decreasing the risk of mental/emotional and social disorders. In 1971, Denton was the primary founder of Atlantic Behavioural Science Applications, an organization for promoting the use of scientifically derived knowledge in the planning, operation, and evaluation of human service programs. On three occasions, his volunteer efforts have been recognized by the Canadian Mental Health Association, the most recent in 1998 when he received the Dr. C.M. Hincks Award for the advancement of mental health. In the same year, he was given recognition in the Nova Scotia Legislature for his service and contribution to the mental health profession. A native Nova Scotian, Dr. Denton lives in Truro, Nova Scotia, where he operates a private practice. PMID- 11381728 TI - Between the laboratory, the school, and the community: the psychology of human development, Toronto, 1916-1956. AB - In the twentieth century, Canadian psychologists have been involved with the educational system and the community at several points in time. In this article, the psychology of human development as developed at the Department of Psychology at the University of Toronto from 1916 to 1956 is investigated. In a variety of projects, the mental health of children was investigated in educational settings while measures were designed and tested to prevent maladjustment and to promote mental health. Initially, research and intervention aimed at adjusting school children to the educational setting. Later, a critical perspective on social institutions and Canadian society was articulated. PMID- 11381729 TI - [Reconnaissance: theoretical construction of social change]. AB - Analysing individual reactions--omnipotence or helplessness--to the question put forward in this symposium, the author observes that the same reactions are present in the mental health field. Investigation of these reactions shows that they alternate in time and are based on a quest for recognition. This quest follows a pattern: First, a group proclaims new knowledge which is based on a fantasy. Then, when this group obtains recognition with its corresponding power, making the transition from a position of impotence to one of power, the fantasy becomes an illusion of omnipotence which excludes others, triggering the cycle again in the excluded groups. This pattern could explain the evolution of the mental health field over the last four decades. Over the last few years, however, there have been some failures in this process, and the author suggests possible solutions to move beyond the impasse. PMID- 11381730 TI - Reminiscences and reflections on mental health services in Ontario. PMID- 11381731 TI - The current mental health system for schizophrenic patients. PMID- 11381732 TI - [Hope and deficiency: community mental health and severe mental disorders]. AB - Over the second half of the twentieth century, the way in which Canadian society treats people with mental challenges has changed radically, from exclusion and containment in asylums to reintegration and maintenance in the community. Now that deinstitutionalization is practically complete, the objective has increasingly shifted from reintegrating people with severe mental challenges in their communities to upholding their rights and ensuring that they have access to resources allowing them to assume their responsibilities and roles as citizens. This change of vision results in a completely different set of issues for community mental health. Given the prejudices surrounding mental illness, the capacity of community mental health professionals to play a mediating and facilitating role becomes a determining factor in successfully reestablishing and re-empowering people with severe mental challenges. The advances that have clearly been made give cause for hope, but in the context of the unwanted effects of deinstitutionalization and unfavourable socioeconomic conditions, significant obstacles still need to be overcome before people with severe mental challenges can participate fully in the life of the collectivity. PMID- 11381734 TI - The emergence of stakeholder contributions. PMID- 11381733 TI - Extended leave. PMID- 11381735 TI - Significant developments in psychiatry: implications for community mental health. PMID- 11381736 TI - "Is that all there is?" Searching for citizenship in the midst of services. PMID- 11381737 TI - [Off-site and alternative services in community mental health care in Quebec]. AB - The objective of this article is to understand the evolution of the practice of "elsewhere" and "otherwise" that alternative community resources in Quebec have consistently advocated. Towards this end, we begin by identifying the meaning and significance of the ideas of "elsewhere" and "otherwise" in the context of the transformation of Quebec's mental health system since 1989. Then we present the results of an exploratory study carried out between 1997 and 1999 in which the coordinators as well as members of 6 community mental health organisations in a semi-urban region of Quebec were interviewed. The results of the study suggest the validity of certain fears that community resources are becoming "deradicalized." The analysis presented here suggests the unfolding of a complex process involving the integration of alternative resources into the broader public mental health system, internal dynamics, and the emerging limitations of the vision of "elsewhere" and "otherwise" that has guided them until now. Twenty years after alternative community resources appeared on the Quebec mental health scene, is it time to reform the philosophy of "elsewhere" and "otherwise"? PMID- 11381738 TI - Suicide prevention in Canada: a history of a community approach. AB - Suicide is a major mental health and public health problem in Canada. Canada's suicide rate ranks above average in comparison to countries around the world. The prevention of suicide predates the European presence in Canada and much can be learned from these endeavours. Current efforts grew largely from the grass roots, with little government support or initiative (with a few provincial/territorial exceptions). Canada's community efforts have been diverse and inclusive. Among such efforts have been: (a) traditional approaches among Native peoples, (b) the establishment of the first crisis centre in Sudbury in the 1960s, (c) the development of a comprehensive model in Alberta, (d) the beginning of a survivor movement in the 1980s, and (e) the national prevention efforts of the Canadian Association for Suicide Prevention. There are, however, striking lacks--most notable among them the paucity of support for research in Canada. Future efforts will call for even greater community response to prevent suicide and to promote wellness. PMID- 11381739 TI - Barriers to recovery in a First Nations community. AB - The practice of psychiatric rehabilitation is a concept and method that developed in urban-based settings. It has become a widely used guiding principle in mental health practice. This research examines how psychiatric rehabilitation fits within a remote First Nations community. Ten people--service providers, consumers, and family members--were interviewed to gather information about their perceptions of and experiences within the mental health system. The interview material was examined using content analysis. The results suggest that geographic and economic factors create serious barriers to application of the psychiatric rehabilitation method in a remote First Nations community. PMID- 11381740 TI - An application of a community psychology approach to dealing with farm stress. AB - Community psychology arose in the 1960s, in the United States, as a response to: (a) pressure to move toward more community based mental health services, and (b) clinical psychologists asking themselves why they were individually treating and conceptualizing large numbers of people who had similar presenting problems. They realized that the social context of their clients' lives was paramount in determining their emotional health. In 1990, I brought a community psychology perspective to designing and implementing a farm stress program in Saskatchewan. I describe how community psychology values and practices, community development, and mental health promotion are applied to this program. I discuss the value of conceptualizing mental health issues, such as farm stress, from the perspective of individuals-in-communities and discuss considerations for future applications of a community psychology approach to similar and other mental health issues. PMID- 11381741 TI - Maternal-fetal pharmacokinetics of methanol. AB - We undertook the present project to elucidate the physiologic factors that govern methanol delivery to the developing conceptus after maternal methanol exposure, and to develop a physiologically based toxicokinetic model to describe methanol disposition in pregnancy. A multi-experimental approach addressed the goals of this project. Initial experiments characterized the systemic disposition of methanol after intravenous or oral administration to nonpregnant female rats. Methanol absorption from the gastrointestinal tract was rapid (peak concentrations appeared within 1 to 2 hours after administration) and essentially complete (systemic bioavailabilities ranged from approximately 0.6 to 1.0). As anticipated for short-chain aliphatic alcohols, methanol elimination from the systemic circulation was nonlinear due to saturation of the metabolic route or routes responsible for converting methanol to formaldehyde and, ultimately, formic acid. However, a significant parallel linear route of methanol elimination was observed, which accounted for an increasingly significant fraction of total elimination as methanol doses (or systemic concentrations) increased. The disposition of methanol after oral or intravenous administration was similar in pregnant and nonpregnant female rats, regardless of the gestational stage (day 7, 14, or 20 after conception) at which the toxicokinetics of methanol were examined. This observation indicated that data from nonpregnant subjects could be used in the development of the maternal portion of a comprehensive physiologic model for methanol disposition. Parallel experiments in female mice indicated that methanol elimination was approximately twice as rapid in mice as in rats due to a significantly higher maximal velocity for methanol metabolism in the smaller rodent species. As was the case in the rat, relatively small changes in methanol elimination were observed during the course of gestation in pregnant mice. In both species, the rate of methanol metabolism by fetal liver in vitro was less than 10% that of the metabolic rate in adult liver. The kinetics of methanol delivery into the fetal environment were examined by determining amniotic fluid concentrations of methanol after intravenous administration to pregnant rats. The net rate of methanol translocation from maternal blood to amniotic fluid decreased as methanol concentration increased. Although the mechanism of this anomalous result is unknown, it possibly is due to a methanol-induced decrease in blood flow to the fetus. PMID- 11381742 TI - DNA adduct formation and T lymphocyte mutation induction in F344 rats implanted with tumorigenic doses of 1,6-dinitropyrene. AB - Diesel emissions are known to induce tumors in laboratory animals and are suspected of being carcinogenic in humans. Of the compounds associated with diesel exhaust, 1,6-dinitropyrene is a particularly potent mutagen and carcinogen; thus, monitoring the toxic effects of 1,6-dinitropyrene may provide a means for assessing the carcinogenic risk associated with exposure to diesel emissions. In these experiments, 1,6-dinitropyrene was implanted into the lungs of rats according to a protocol known to induce lung tumors; the DNA adducts were characterized and quantified in target (lung) and surrogate (liver, white blood cells, and spleen lymphocytes) tissues. In addition, mutation induction was assayed at the hypoxanthine-guanine phosphoribosyltransferase (hprt) locus in spleen T lymphocytes, and the relation between adduct concentration and mutation induction was elucidated. Rats were administered 30 micrograms or 100 micrograms of [ring-3H]1,6-dinitropyrene and adduct levels were quantified for up to one month after treatment. In lung tissue, white blood cells, liver tissue, and spleen lymphocytes, one major DNA adduct, N-(deoxyguanosin-8-yl)-1-amino-6 nitropyrene, was detected. In each tissue, the levels of this adduct reached a maximal level one to seven days after treatment and decreased to approximately 25% of the peak values by 28 days after treatment. DNA adduct formation in the lung was approximately tenfold higher than that observed in the other tissues. A dose-response relation was not observed in the lung, white blood cells, or liver; but in the spleen lymphocytes, a threefold increase in dose resulted in approximately a twofold increase in adduct formation. After a similar treatment with 1,6-dinitropyrene, mutations were assayed at the hprt locus in spleen T lymphocytes for up to 51 weeks. Compared with control animals treated with solvent, 1,6-dinitropyrene induced a significant increase in mutant frequency with the 100-microgram dose, typically producing twofold more mutants than the 30 microgram dose. With both doses, the mutant frequency increased until 21 weeks after treatment with 1,6-dinitropyrene, remained constant until week 40, and then began to decrease. Nonetheless, nearly one year after treatment, the mutant frequency in rats treated with 1,6-dinitropyrene was greater than that observed in control rats. In a subsequent experiment, rats were administered 0, 0.3, 1, 3, 10, 30, 100, or 150 micrograms of 1,6-dinitropyrene and the extent of adduct formation was determined seven days after treatment. In the lung nuclei, liver nuclei, and spleen lymphocyte DNA, a significant dose-response relation was observed, with the extent of adduct formation increasing significantly at a dose of 10 micrograms. A twofold increase in dose resulted in a twofold increase in adduct formation up to the 30-microgram dose in lung nuclei DNA, and up to the 10 microgram dose in liver nuclei DNA. At higher doses, the extent of adduct formation still increased but the rate of increase was much lower than that occurring at lower doses. To assess the mutation frequency as a function of dose, additional rats were treated with 0, 0.3, 1, 3, 10, 30, 100, or 150 micrograms of 1,6-dinitropyrene and mutations were assayed at the hprt locus in spleen T lymphocytes 21 weeks after treatment. In this experiment, a significant dose response relation was observed, with the increase in mutants becoming significant at 1 microgram and higher doses of 1,6-dinitropyrene. These data indicate that 1,6-dinitropyrene, a constituent of diesel emissions, is metabolically activated by nitroreduction to produce DNA adducts in target and surrogate tissues. They further suggest that T lymphocyte mutations may be a more sensitive and longer lived biomarker than DNA adducts for assessing previous exposures to genotoxic agents, such as nitro-polynuclear aromatic hydrocarbons. PMID- 11381743 TI - Oil and seabirds--the imperative for preventing and reducing the continued illegal oiling of the seas by ships. PMID- 11381744 TI - Cancer drugs from microalgae. PMID- 11381745 TI - Charging for port reception facilities in North Sea ports: putting theory into practice. AB - The aim of this paper is to evaluate the charging systems for the use of port reception facilities for waste oil, and to examine the potential impact of the charging elements of the new (late 2000) EU Directive on port reception facilities for ship-generated waste and cargo residues. Experience to date with alternative models for charging is considered. Conclusions are drawn about the effectiveness of the EU Directive as a means of controlling pollution in the North Sea and producing a 'level playing field' between ports. PMID- 11381746 TI - Box coring artifacts in sediments affected by a waste water outfall. AB - Variations in porosity of surface sediments are often the major cause of sediment loss during gravity and box coring. Sediments with a high content of organic matter usually have higher porosity, and thus, lower resistance (strain) towards mechanical disturbance. Here, we demonstrate that box coring artifacts (i.e. sediment loss and core shortening) can be produced in sediments from the Palos Verdes (PV) shelf, which in the past had received relatively high loads of organic carbon (OC) enriched particulate matter originating from the Whites Point outfall that had created a high porosity layer at depth. This has been overlooked as a possibility for obtaining low estimates of sediment and pollutant accumulation rates. Since any such sediment loss during coring can lead to serious underestimates of sedimentation rates, our results here may have important implications for any attempts at reconstructing pollutant fluxes and histories in these coastal marine sediments. PMID- 11381747 TI - Contamination of polychlorinated biphenyls (PCBs) in sediments from Kyeonggi Bay and nearby areas, Korea. AB - To elucidate the characteristic distribution and contamination of polychlorinated biphenyls (PCBs) in sediments, 63 sediments and five benthic organisms from Kyeonggi Bay, Namyang Bay and Lake Shihwa, West coast of Korea, were analysed. Characterization of PCBs distribution in sediments was conducted by correlation between PCBs concentrations and environmental parameters, comparison of contamination level and composition of PCBs homologs between Kanechlor mixture (KC-mix) and sediments. The residues of PCBs in sediments were correlated with total organic carbon (TOC) and particulate organic carbon (POC) contents, not with mud contents and grain size distributions. Elevated concentrations of T-PCBs were found in sediments from Incheon North Harbor (INH) in Kyeonggi Bay. T-PCBs concentrations were decreased with distance increase from inner site of INH. The residues of T-PCBs in sediments from Namyang Bay were either non-detectable or near to detection limit. The contamination by PCBs in sediments from Lake Shihwa was also low. The PCBs congener profiles in INH were similar to those of KC-mix, while those in less contaminated sites showed relatively high percentage of lower chlorinated biphenyls. Sites K18 (580 ng/g or 48 ug/g-OC dry wt) and K19 (330 ng/g or 38 ug/g-OC dry wt) within INH exceeded the sediment quality criteria (SQC) (16 micrograms/g-OC as a KC-mix) derived from equilibrium partitioning (EqP) approach. PMID- 11381748 TI - PCB concentrations and metabolism patterns in common terns (Sterna hirundo) from different breeding colonies in The Netherlands. AB - PCB levels in blood of common terns (Sterna hirundo) from Terneuzen, a breeding colony in The Netherlands with relatively low breeding success, were significantly higher than in birds from two Dutch reference colonies, one nearby (Oesterdam) and one further away (the Isle of Griend). However, a detailed analysis of the patterns of PCB congeners in birds from the different colonies indicated that birds from Terneuzen and to a lesser extent birds from the Oesterdam showed Ah-receptor mediated activity of P450 iso-enzymes. It is concluded that the induction of this enzyme activity in Terneuzen may be related to PCBs as well as other non-identified compounds. In contrast to this, it is likely that at the Oesterdam non-identified compounds are mainly responsible for the enzyme activity, resulting in a shift of PCB congener patterns. PMID- 11381749 TI - Assessment and impact of microbial fecal pollution and human enteric pathogens in a coastal community. AB - The goals of this study were to assess watersheds impacted by high densities of OSDS (onsite sewage disposal systems) for evidence of fecal contamination and evaluate the occurrence of human pathogens in coastal waters off west Florida. Eleven stations (representing six watersheds) were intensively sampled for microbial indicators of fecal pollution (fecal coliform bacteria, enterococci, Clostridium perfringens and coliphage) and the human enteric pathogens, Cryptosporidium, Giardia, and enteroviruses during the summer rainy season (May September 1996). Levels of all indicators ranged between < 5 and > 4000 CFU/100 ml. Cryptosporidium and Giardia were detected infrequently (6.8% and 2.3% of samples tested positive, respectively). Conversely, infectious enteroviruses were detected at low levels in 5 of the 6 watersheds sampled. Using cluster analysis, sites were grouped into two categories, high and low risks, based on combined levels of indicators. These results suggest that stations of highest pollution risk were located within areas of high OSDS densities. Furthermore, data indicate a subsurface transport of contaminated water to surface waters. The high prevalence of enteroviruses throughout the study area suggests a chronic pollution problem and potential risk to recreational swimmers in and around Sarasota Bay. PMID- 11381750 TI - Can a pollution event be detected using a single biological effects monitoring method? AB - The trends of malformation prevalence in embryos of dab, Limanda limanda, in the southern North Sea after the year 1990 mirrored the drop in major pollutants in the rivers draining into the German Bight. Despite this general decline, we detected a pollution event in the southern North Sea in winter 1995/1996 employing the prevalence of malformations in pelagic dab embryos as an indicator. An abrupt rise in malformation prevalence in the embryos of dab, corresponded to a dramatic increase in DDT levels in parent fish from the same area, indicating a hitherto unnoticed introduction of considerable quantities of DDT into the system. PMID- 11381751 TI - An evaluation of marine bird population trends following the Exxon Valdez oil spill, Prince William Sound, Alaska. AB - We examined post-spill trends (1989-1998) of marine bird populations in Prince William Sound (PWS) following the Exxon Valdez oil spill (EVOS) to evaluate recovery of injured taxa. Two criteria were employed. First, we examined population trends of injured taxa only in the oiled area of PWS using regression models. Second, we examined population trends of injured taxa in the oiled area relative to the unoiled area using homogeneity of the slopes tests. We considered a population recovering if there was a positive trend using either criteria. We considered a population not recovering if there was no trend using either criteria or a negative trend in the oiled area. A significant negative trend in the oiled area relative to the unoiled area was considered a continuing and increasing effect. Most taxa for which injury was previously demonstrated were not recovering and some taxa showed evidence of increasing effects nine years after the oil spill. Four taxa (loons Gavia spp, Harlequin Duck Histrionicus histrionicus, Bufflehead Bucephala spp, and North-western Crow Corvus caurinus) showed weak to very weak evidence of recovery. None of these taxa showed positive trends in both winter and summer. Nine taxa (grebes Podiceps spp, cormorants Phalacrocorax spp, Black Oystercatcher Haematopus bachmani, Mew Gull Larus canus, Glaucous-winged Gull Larus glaucescens, terns Sterna spp, murres Uria spp, Pigeon Guillemot Cepphus columba, and murrelets Brachyramphus spp) showed no evidence of recovery during summer or winter. Four taxa (scoters Melanitta spp, mergansers Mergus spp, goldeneyes Bucephala spp, and Black-legged Kittiwaka Rissa tridactyla) showed evidence of continuing, increasing effects. We showed evidence of slow recovery, lack of recovery, and divergent population trends in many taxa which utilize shoreline and nearshore habitats where oil is likely to persist. Potential lingering spill effects and natural variability appear to be acting in concert in delaying recovery of many PWS bird populations. PMID- 11381752 TI - Toxicity of chromated copper arsenate (CCA)-treated wood to non-target marine fouling communities in Langstone Harbour, Portsmouth, UK. AB - The effect of the anti-marine-borer timber preservative chromated copper arsenate (CCA) (a pressure impregnated solution of copper, chromium and arsenic compounds) on non-target marine fouling animals was investigated during a subtidal exposure trial. Panels of Scots pine treated to target retentions of 12, 24 and 48 kg CCA per m-3 of wood, plus untreated controls were submerged at a coastal site on the south coast of the UK for 6, 12 and 18 months. After each exposure period the fouling communities that formed on the surface of panels were assessed both qualitatively and quantitatively. Community structure was similar on panels treated to the three CCA loadings, but was significantly different from community structure on untreated panels. The total number of species (species richness) was similar on all panels, although the number of individual organisms attached to the surface of panels was significantly higher on CCA-treated panels than on untreated panels. k-dominance curves revealed that the difference in numbers of individuals between CCA-treated and untreated panels was caused by higher numbers of the dominant species (Elminius modestus, Hydroides ezoensis, and Electra pilosa) on CCA-treated panels. Other species were present in similar numbers on panels of all treatments. Results indicate that there are no detrimental toxic effects to epibiota caused by the presence of CCA preservative within the matrix of the wood at any of the treatment levels. Differences in community structure between CCA-treated and untreated panels may be due to enhanced larval settlement on CCA-treated timber by some species as a result of modifications to the surface properties of the timber by the CCA preservative. PMID- 11381753 TI - The problem of transfer of radionuclide pollution by sea ice. AB - The problem of the radioactive pollution of the ice cover is discussed in this paper. An estimation of the possible transport of the radionuclides by drifting ice from the Kara Sea into the Arctic Ocean is presented. The conclusion about the importance of the contribution of drifting ice to the contamination of the Greenland coastal water is based on the comparison of estimates of the multiyear export of radioactive drifting ice from the Kara Sea with the observed concentrations of the radionuclides in the Greenland coastal water. PMID- 11381754 TI - Genotoxic hazard of pollutants in cetaceans: DNA damage and repair evaluated in the bottlenose dolphin (Tursiops truncatus) by the Comet Assay. AB - Single cell gel electrophoresis (or Comet Assay) was used for evaluation of the in vitro genotoxicity of hydrogen peroxide (used as a positive control), polychlorinated biphenyls (PCBs) (Aroclor 1254) and methyl mercury chloride, in isolated bottlenose dolphin leukocytes. Results showed that hydrogen peroxide and methyl mercury induced DNA strand breakage in a dose-dependent manner, while PCBs did not induce a clear dose-effect response at the low doses investigated. Efficiency in repairing DNA breakage induced by methyl mercury was also evaluated. Findings demonstrated that dolphin cells are characterized by higher efficiency in DNA repair when compared to human leukocytes. The observed resistance to methyl mercury toxicity in dolphins was hypothesized to be a defence strategy developed to combat high dietary exposure and compensate for limited capacity to excrete persistent pollutants. PMID- 11381755 TI - Methylmercury in molluscs along the French coast. PMID- 11381756 TI - [Hip arthroscopy]. PMID- 11381757 TI - [Comparison of 2 surgical techniques in isolated supraspinatus rupture. A matched pair study]. AB - The comparison of results after open surgery methods in the repair of rotator cuff ruptures needs homogen patient groups. We compared 34 patients in 2 groups who were treated with open surgery for isolated ruptures of the supraspinatus muscles (lesion type III). Patients in group I were treated with transosseous suture refixation of the supraspinatus, in group II suture anchors (type: Corkscrew) were used. Both groups were age and gender paired. Preoperatively, all patients had the same clinical status. For the evaluation of the clinical results at follow-up we used the Constant-Score. In addition, strength and pain measurement and oblique X-rays of the shoulder were performed. The follow-up time was 18 months. In all patients, the mean Constant-Score improved significantly with increase in muscle strength and marked pain reduction. Significant differences between both groups could not be observed. The mean time of surgery in group II (suture anchor) was significantly lower than in group I. In conclusion, with the anchor techniques the same good results can be achieved as with bone refixation. The shortening of the surgery time should, however, be seen in relation to the higher costs with this technique. PMID- 11381758 TI - [Hip arthroscopy. Minimal invasive diagnosis and therapy of the diseased or injured hip joint]. AB - Arthroscopy of the hip joint has developed into a useful tool for the hip surgeon. Hip joint anatomy, however, makes special demands of the arthroscopist. He needs to be familiar with the arthroscopic anatomy of the hip and its variations. Moreover, he should have practical training in the technique of hip arthroscopy prior to his first intraoperative experience in order to avoid complications. A complete arthroscopic inspection of the hip can be achieved by using a combined procedure: whereas the central hip compartment can be scoped only by distraction of the joint, the periphery can be better seen without traction. Whether to place the patient supine or lateral is dependent on personal experience. No matter which position is used, the positioning technique has to be exact. The literature has shown that most complications are related to traction. Before the first portal is placed, the joint vacuum force should be broken by distension of air or fluid. This leads to maximum distraction of the joint and reduces the risks of damage to labrum and cartilage during first access to the joint. For a diagnostic round through the central compartment, at least two portals have to be placed. The use of a 3-portal technique increases the range of inspection. Due to the relatively thin soft tissue mantle and greater distance to neurovascular structures, the anterolateral or lateral portal should be used as the first portals to the central compartment. In addition, the anterolateral portal is the standard portal to the periphery of the hip. The posterolateral or anterior portal should be used as a supplementary portal. The following indications have been described for an arthroscopic procedure of the hip: loose bodies, labral lesions, synovial diseases such as chondromatosis and pigmented villonodular synovitis, associated lesions in underlying osteoarthritis, ruptures of the teres ligament, malorientation of the acetabulum and proximal femur and, last but not least, "idiopathic" hip pain. The use of hip arthroscopy in infectious arthritis, avascular necrosis of the femoral head, Perthes' disease, osteochondrosis dissecans and complications after total hip replacement is less frequent. Here, in addition to its diagnostic value, operative arthroscopy of the hip offers removal of loose bodies, resection of the labrum and ligaments, synovial biopsy, partial synovectomy, microfracturing, lavage and placement of intraarticular drainage. The first results of arthroscopic procedures in the hip are promising. In addition to its diagnostic value and contribution to the understanding of intraarticular anatomy and pathology, recent studies have demonstrated the advantages of the arthroscopic treatment of the hip. PMID- 11381759 TI - [Comparative biomechanical compression trials with a new vertebral prosthetic implant]. AB - The authors present a new titanium implant for replacement of the vertebral body (Synex). Possible indications would be fractures or dislocations with destruction of the anterior column, posttraumatic kyphosis as well as tumors in the throracolumbar spine. The construction has to be completed by a stabilizing implant. For best fit and contact to adjacent end-plates Synex is distractable in situ. The possibility of secondary dislocation or loss of correction should thereby be minimised. OBJECTIVES: We performed comparative compression tests with Synex and MOSS ("Harms mesh cage") on human cadaveric specimens of intact vertebrae (L1). The aim of the study was to measure the compressive strength of the vertebral body end-plate in uniaxial loading via both implants to exclude a caving of Synex in vivo. METHODS: 12 human cadaveric specimens of intact vertebrae (L1) were divided in 2 similar groups (matched pairs) according to bone mineral density (BMD), determined using DE-QCT. The specimens were loaded with axial compression force at a constant speed of 5 mm/min to failure and the displacement was recorded with a continuous load-displacement curve. RESULTS: The mean ultimate compression force (Fmax) showed a tendency towards a higher result testing Synex with 3396 N versus 2719 N (non significant). The displacement until Fmax was 2.9 mm in group S (Synex), which was half as long as in group M (5.8 mm). The difference was significant (p < 0.001). The compression force was twice as high and significantly (p < 0.05) higher with Synex at a displacement of 1 mm, 1.5 mm and 2 mm. A significant (p < 0.001) correlation (R = 0.89) between Fmax and BMD was found. CONCLUSIONS: Synex was found to be at least comparable to MOSS for suspensory replacement of the vertebral body at the thoracolumbar spine. A possible consequence of the significantly higher mean compression forces between 1 and 2 mm displacement might be a decreased segmental deformation or loss of correction. PMID- 11381760 TI - [Correction of malunited distal radius fractures by osteotomy and interposition of iliac crest bone spain in the radio-palmar position. A retrospective study of 42 patients]. AB - The retrospective study included 42 patients who have been treated with a palmar opening wedge osteotomy after malunion of distal radius fracture. 31 patients could be reexamined clinically and radiologically. The corrective osteotomy on the distal radius was done only radiopalmar with an insertion of a corticocancellous iliac graft. Radiological and functional findings as well as the subjective judgement of the patient have been commented pre- and after surgically according to the assessment scheme of Pechlaner and were then compared with the contra lateral wrist. The evaluation of the x-ray pictures showed an improvement from 3.7 to 1.9 points, the functional improvement from 3.7 to 2.6 points. The improvement of the subjective complaints results in a diminution of 3.3 to 2.2 points. PMID- 11381761 TI - [Callus development after bored and unbored femoral interlocking nailing. An experimental study]. AB - This was a prospective randomized baboon animal study, the study protocol was approved by the ethical committee according to the international guidelines for animal research projects. In 8 animals a midschaft femoral osteotomy was stabilized with reamed femoral interlocking nailing and in 8 animals by unreamed locked femoral nailing. Polychrome sequence bone labeling was done 5 weeks postop. with calcein-green, 8 weeks postop. with xylenol-orange and 10 weeks postop. with oxytetracycline. 10 weeks postop. the animals were sacrificed, the femurs explanted and planimetric and epifluorescence histomorphometric evaluation of serial transverse sections were done. In planimetric histomorphometric evaluation in unreamed femoral nailing a mean endostal callus formation was recorded with 28.0 +/- 9.9 mm2 per section and in reamed femoral nailing with 11.5 +/- 5.0 mm2 (p < 0.001). Periostal callus formation was recorded in the unreamed group with 238.7 +/- 87.1 mm2 per section and in the reamed group with 142.1 +/- 71.9 mm2 (p < 0.001). In epifluorescence histological evaluation endostal as well as periostal callus formation was more extensive and earlier after unreamed than reamed femoral nailing. Endostal callus formation was found in all animals after unreamed femoral nailing, and was present in 2 out of 8 specimen in the reamed group. Also 1 out of 8 animals in the reamed group developed a non-union. Unreamed femoral nailing with low diameter interlocking nails proved to be safe regarding bone healing in this experimental model with obvious advantages both in amount and time course of callus formation compared to reamed femoral nailing. Based on this results unreamed femoral nailing techniques can be recommended for femoral fractures. PMID- 11381762 TI - [Osteosynthesis of fractures of the head of the tibia in advanced age. A matched pair analysis]. AB - In a retrospective study we analyzed 183 patients with tibial head fractures. 15 patients were older than 60 years at the time of accident, with a mean age of 69 (62-81) years. All patients had sustained an isolated tibial plateau fracture with a split-impression fracture in 10 patients, an impression fracture in 2 patients and a bicondylar fracture in 3 patients. The mean time to follow-up was 5.1 years. In a matched-pair-analysis this group was compared to a group of young patients with a mean age of 43 years (26-56) and identical lesions and surgical procedures. The mean time to follow-up in this group was 4.2 years. We observed that fractures at higher age resulted mostly from trivial accidents. Except from a prolonged inpatient time the initial course was comparable. In a 66 year-old woman early implant removal was necessary because of infection. Despite identical surgical treatment we observed a loss of reduction with incongruency of the plateau and deviation of the axis. Correspondingly older patients displayed a statistically significant threefold increase of osteoarthritis compared to younger patients. However, there was no clinical correlation using the Rasmussen, OAK and Lysholm score. Both groups displayed no statistical significant differences for the parameters function, stability and pain. According to the old age the only significant difference was a reduction of the Tegner score. Reduction and internal fixation of tibial plateau fractures in an elderly population displayed mostly a satisfactory clinical result within this 5-year period. The problem of loss of reduction and rapid osteoarthritis remains to be evaluated in a larger group over a longer period of time. PMID- 11381763 TI - [Quality of life of severely injured patients 1 year after trauma. A matched-pair study compared with a healthy control group]. AB - PURPOSE: Severe trauma is accompanied not only with functional disabilities, but also with pain, social and psychological problems of the patient. Those four aspects are important components of the construct quality of life (QoL). The purpose of this study was to evaluate different questionnaires and to compare results between multiply injured patients and healthy controls. METHODS: A group of 43 severely injured patients was matched according to age, gender, educational status and family life situation to 43 persons who had never experienced a severe injury. For the assessment of all four aspects of QoL the patients completed validated questionnaires: Beck-Depression-Inventory (BDI), Short Form Health Survey (SF-36), State-Trait Anxiety Inventory (STAI), a questionnaire for locus of control (KKG), and a questionnaire for social support (SOZU). RESULTS: Patients and the control group showed remarkable differences: 22 of 43 trauma patients suffered from bad health perception (subscale of SF-36) (8/43 control persons), 21 trauma patients reported moderate to severe pain (5/43 control persons), 11 of 43 trauma patients suffered from anxiety (4/43 control persons) and 10 of 43 trauma patients complained about depressive symptoms (2/43 control persons). CONCLUSION: One year after trauma, patients suffer from severe impairments, some even in all four aspects of QoL. The questionnaires used are very well applicable to patients after severe injuries. A trauma-specific Quality of Life Instrument is lacking--and is currently developed by the german trauma registry group. PMID- 11381764 TI - [Strength testing of pins made of human femur cortical bone]. AB - For biomechanical evaluation of cortical bone as a potential source for internal fixation devices, pins of 2 and 3 mm in diameter were made from fresh human femora and subjected to bending loads. Some of the pins were tested without further treatment; others were defatted and dried in acetone solution. Of the acetone-treated pins, some were autoclaved either at 121 degrees C for 20 min or at 134 degrees C for 8 min. Acetone treatment of the pins caused a 5.5% increase in bending strength (p < or = 0.05) and a 6.6% increase in the elastic modulus (p < or = 0.05). The mean bending strength of the acetone-treated pins was 257 +/- 52 N/mm2 and the mean elastic modulus was 18,346 +/- 1292 N/mm2. Autoclaving of the pins at the lower temperature for a longer time led to a decrease in the bending strength of up to 16% and at the higher temperature for a shorter time to a decrease of up to 41%. To compare the strengths of the cortical and synthetic pins, 2-mm Biofix pins were also tested. The Biofix pins showed a 25% higher bending strength than the acetone-treated pins but a 12.5% lower elastic modulus. It should be noted that in animal studies the synthetic Biofix pins showed a 20% loss of strength within 2 days after implantation in situ. PMID- 11381765 TI - [Clavicular fractures]. PMID- 11381766 TI - [Severe complications of Staphylococcus aureus infection in the child]. AB - Life threatening complications can be caused by staphylococcal infection, even if they appear as a simple pustulosis, via hamatogenous dissemination. On the other hand the bacteria is able to penetrate the skin without pre-existing skin lesions and reach the adjacent soft tissue, as was the case with a 9 year old boy who had a contusion of the left shoulder and a consecutive life threatening septicemia. PMID- 11381767 TI - [Traumatic hemipelvectomy and postoperative quality of life. Report of 2 cases 18 years after trauma]. AB - Report and progress of 2 traumatic hemipelvectomies in motor-cyclists with excluding extraperitoneal tearing of half of the pelvis and disrupture of the pelvic vessels for 18 years. The purpose of this case study was the analysis of the quality of life, the clinical complications and the reintegration of the 2 patients 18 years after traumatic hemipelvectomy. PMID- 11381768 TI - [Post-traumatic rotational deformity of the femur in a patient with paraplegia]. AB - A 37-year old woman with paraplegia sustained a posttraumatic rotational deformity of the femur of 40 degrees after the osteosynthesis of a femur fracture. In the consequence she got several problems in her everyday life, such as problems in handling the wheel-chair and in catheterism. The determination of anteversion of the femoral neck by computed tomography is necessary for quantifying rotational faults and planning of corrective osteotomy. Our patient recuperated as the same level of independence after a subtrochanteric corrective osteotomy as before the fracture of the femur. PMID- 11381769 TI - [Abdominal compartment syndrome. Tons C, Schachtrupp A, Trau, M, Mumme T, Schuplelick V (2000). Abdominal compartment syndrome: prevention and treatment. Chirurg 71: 918-926]. PMID- 11381770 TI - Soil, water and nutrient conservation in mountain farming systems: case-study from the Sikkim Himalaya. AB - The Khanikhola watershed in Sikkim is agrarian with about 50% area under rain-fed agriculture representing the conditions of the middle mountains all over the Himalaya. The study was conducted to assess overland flow, soil loss and subsequent nutrient losses from different land uses in the watershed, and identify biotechnological inputs for management of mountain farming systems. Overland flow, soil and nutrient losses were very high from open agricultural (cropped) fields compared to other land uses, and more than 72% of nutrient losses were attributable to agriculture land use. Forests and large cardamom agroforestry conserved more soil compared to other land uses. Interventions, like cultivation of broom grass upon terrace risers, N2-fixing Albizia trees for maintenance of soil fertility and plantation of horticulture trees, have reduced the soil loss (by 22%). Soil and water conservation values (> 80%) of both large cardamom and broom grass were higher compared to other crops. Use of N2-fixing Albizia tree in large cardamom agroforestry and croplands contributed to soil fertility, and increased productivity and yield. Bio-composting of farm resources ensured increase in nutrient availability specially phosphorus in cropped areas. Agricultural practices in mountain areas should be strengthened with more agroforestry components, and cash crops like large cardamom and broom grass in agroforestry provide high economic return and are hydroecologically sustainable. PMID- 11381771 TI - Progress with implementing local air-quality management in rural areas of England. AB - Air quality in the UK, although vastly improved from the smogs of the 1950s, now faces a new set of challenges from a variety of sources and pollutants. Poor air quality has long been associated with urban areas, but it is becoming clear that many rural locations also have locations likely to exceed the UK Air Quality Strategy-objectives: This paper will examine the extent to which rural authorities have been engaged in the local air-quality management (LAQM) process, a new regime by which air quality control is being accomplished in the UK. Results are presented from a questionnaire survey of environmental health officers of 100 rural authorities undertaken in January 1999. The paper investigates both the technical aspects of the LAQM process as well as the management approaches. The current progress of rural authorities, and some of the problems they face, are discussed particularly in comparison with urban areas. It is concluded that some rural authorities with air pollution problems stemming either from within or outwith their borough, may face significant challenges from the LAQM review and assessment process, particularly where air pollution responsibilities have only relatively recently been addressed. Rural authorities seem to be embracing these new responsibilities enthusiastically, but it is almost inevitable that they will be trailing behind urban authorities who have several decades of experience and joint working to draw upon. PMID- 11381772 TI - Economics of reservoir sedimentation and sustainable management of dams. AB - Accepted practice has been to design and operate reservoirs to fill with sediment, generating benefits from remaining storage over a finite period of time. The consequences of sedimentation and project abandonment are left to the future. This 'future' has already arrived for many existing reservoirs and most others will eventually experience a similar fate, thereby imposing substantial costs on society. Such costs could be avoided if sedimentation was minimized and dams were allowed to live forever. The fact that the world's inventory of suitable reservoir sites is limited provides an additional reason for encouraging the sustainable management of dams. This paper provides a framework for assessing the economic feasibility of sediment management strategies that would allow the life of dams to be prolonged indefinitely. Even if reduced accumulation or removal of sediment is technically possible, its economic viability is likely to depend on physical, hydrological and financial parameters. The model presented incorporates such factors and allows a characterization of conditions under which sustainable management would be desirable. The empirical implementation of the model draws upon the substantial amount of technical information available. We analyze the sustainability of reservoirs, with a focus on the trade-off between such sustainability and the short to medium term benefits which a reservoir is expected to produce. The results show that, for a very wide range of realistic parameter values, sustainable management of reservoirs is economically more desirable than the prevailing practice of forcing a finite reservoir life through excessive sediment accumulation. PMID- 11381773 TI - Indirect carbon reduction by residential vegetation and planting strategies in Chicago, USA. AB - Concern about climate change has evoked interest in the potential for urban vegetation to help reduce the levels of atmospheric carbon. This study applied computer simulations to try to quantify the modifying effects of existing vegetation on the indirect reduction of atmospheric carbon for two residential neighborhoods in north-west Chicago. The effects of shading, evapotranspiration, and windspeed reduction were considered and were found to have decreased carbon emissions by 3.2 to 3.9% per year for building types in study block 1 where tree cover was 33%, and -0.2 to 3.8% in block 2 where tree cover was 11%. This resulted in a total annual reduction of carbon emission averaging 158.7 (+/- 12.8) kg per residence in block 1 and 18.1 (+/- 5.4) kg per residence in block 2. Windspeed reduction greatly contributed to the decrease of carbon emission. However, shading increased annual carbon emission from the combined change in heating and cooling energy use due to many trees in the wrong locations, which increase heating energy use during the winter. The increase of carbon emission from shading is somewhat specific to Chicago, due in part to the large amount of clean, nuclear-generated cooling energy and the long heating season. In Chicago, heating energy is required for about eight months from October to May and cooling energy is used for the remaining 4 months from June to September. If fossil fuels had been the primary source for cooling energy and the heating season had been shorter, the shading effects on the reduction of carbon emission would be greater. Planting of large trees close to the west wall of buildings, dense planting on the north, and avoidance of planting on the south are recommended to maximize indirect carbon reduction by residential vegetation, in Chicago and other mid and high-latitude cities with long heating seasons. PMID- 11381774 TI - The consequences of tourism for sustainable water use on a tropical island: Zanzibar, Tanzania. AB - Many developing countries in the tropics have focused on tourism to generate additional income sources and to diversity the economy. Coastlines in particular have been on the forefront of tourist infrastructure development. Here, the presence of a large number of tourists has often had negative consequences for the sustainable use of the available resources, which in turn has had an effect on the integrity of the ecosystems. In this paper, the situation is described for the use of freshwater resources on the east coast of Zanzibar, Tanzania. This region is water poor, relying on freshwater derived from seasonal rains and stored in less efficient aquifers, which consist of freshwater lenses floating on the underlying seawater. Tourism in the area has grown rapidly in recent years and is expected to further increase in the future. This development is expected to put additional pressure on the freshwater resources of the east coast, which show already signs of over-use. The consequences of overexploitation can include the lowering of the groundwater table, land subsidence, deteriorating groundwater quality, and saltwater intrusion. These, in turn, determine the living conditions in coastal areas and the effects will be felt both by the local populations and the tourist industry. An investigation is made into the causes and consequences of water abstraction by the tourist industry. The results show that present levels of withdrawal are not sustainable, and parts of the local populations are already experiencing water deficits on a daily basis. In the future, if the expected increase in tourist numbers occurs, the pressure on the aquifers will correspondingly increase. The results could be that the tourism in the area becomes unsustainable, which could have an adverse effect on the national economy and also on the local population and environment. Therefore, a precautionary water-management approach is suggested. PMID- 11381775 TI - Urinary tract infections during pregnancy in South Carolina. AB - This study presents evidence that over 20 percent of pregnant women with a UTI in South Carolina did not have an antibiotic pharmacy claim within 14 days of diagnosis. Untreated maternal UTI in pregnancy was associated with a 22 percent increased risk for MR/DD in the infant compared to the risk for women who had a UTI and a pharmacy claim for an antibiotic and 31 percent increased risk compared to women who did not have a UTI. The importance of medications compliance should be emphasized in the care of pregnant women. PMID- 11381776 TI - Cardiac amyloidosis. AB - Cardiac amyloidosis should be considered in a patient with heart failure, who is normotensive with decreased left ventricular systolic function and marked left ventricular hypertrophy by echocardiogram and has decreased voltage by ECG. Furthermore, when the diagnosis of cardiac amyloid is made, it is important to classify the subtype of disease to be able to offer appropriate treatment. Contrary to traditional belief that the prognosis for patients with amyloidosis is dismal, some forms of this disease are curable and other forms are characterized by slow progression of disease. PMID- 11381777 TI - Prediction of non-occurrence of syncope during a tilt-table test by early heart rate variations. AB - A recent study showed that, in a select patient population with no drug use and no cardiac or other illness, an increase in heart rate equal to or less than 18 beats per minute from baseline in the first 6 minutes of a tilt-table test at 60 degrees identifies patients who will not develop syncope during prolonged tilting, with specificity and positive predictive value nearing 100 percent. We retrospectively reviewed 110 consecutive tilt-table tests at an angle of 70 degrees or more, performed at our institutions between 1994 and 1999 in patients with and without cardiac disease or drug use. Excluded were 320 additional patients due to either incomplete heart rate documentation or development of syncope in the first ten minutes of tilting. The difference between maximal heart rate in the first ten minutes during tilting and the average of at least two baseline heart rate measurements was used to assess correlation with the non occurrence of syncope during the same test. The tilt-table angle used varied from 70 degrees to 90 degrees (80 degrees in 89 percent of patients). There was a strongly significant (p < 0.0001) correlation between a sustained rise in heart rate equal to or less than 18 beats per minute in the first ten minutes of tilting and the non-occurrence of subsequent syncope during the test. The sensitivity, specificity, positive and negative predictive values were 75.7 percent, 65.0 percent, 79.1 percent and 60.47 percent, respectively. These data indicate that even in the routine setting, that is, a non-select population and at a higher tilt-table angle, an increase in heart rate equal to or less than 18 beats per minute in the first ten minutes of tilting constitutes a good predictor of a negative test. However, the specificity and positive predictive value of this criterion were not high enough to justify an early termination of the test. PMID- 11381778 TI - The status of injury in upstate South Carolina. AB - In conclusion, injuries are a leading cause of emergency room visits and hospital admissions in the upstate of South Carolina. Age was a major factor in the type of injury risk, hospitalization and, once hospitalized, length of stay and cost of hospital care. Future research efforts should include both quantitative and qualitative approaches to develop a more precise profile of persons who are most at risk for injury due to falls, motor vehicle crashes and intentional injuries. Information is also needed on follow-up care, including the cost of care post hospital discharge and after discharge from the emergency room. The recommendations offered in this report may provide health care providers and health care agencies in the four-county upstate region with a guide to begin examining the major types of injuries that occur within their respective communities. Moreover, populations that are disproportionately affected may be delineated, and interventions may be specifically designed for and implemented in partnership with these populations. PMID- 11381779 TI - Multivariate correlations of geochemical binding phases of heavy metals in contaminated river sediment. AB - Distributions of geochemical binding phases of seven heavy metals (Cr, Cu, Co, Zn, Ni, Pb, and Cd) in sediment cores taken from six heavily polluted sites of the Ell-Ren River in Southern Taiwan were studied. Sequential extraction procedures (SEP) were used to determine the variations of heavy metal binding phases (exchangeable, bound to carbonates, bound to manganese-oxides, bound to iron-oxides, and bound to organic matter) in different sediment depths. Multivariate analyses were used to explore the correlations among these geochemical binding phases of heavy metals. Results showed that the total amounts of various binding phases of heavy metals significantly varied with sediment depth, but their binding behaviors in various phases did not significantly change with depth. The organic matter content in the sediments increased with increasing Fe-oxide content. In addition, the binding affinities of carbonates with Zn, Pb, and Ni were higher than the affinities of carbonates with the other heavy metals. The binding affinity of Fe-oxides with Cr was higher than the affinities of Fe oxides with the other heavy metals. Both correlation matrixes and principal component analyses demonstrated that distributions of Cu, Zn, Ni, and Cd had significant correlations with each other in both different depth horizons and various geochemical binding phases. The results indicate that these heavy metals might be discharged from the same pollution sources in the past, and also showed stable geochemical binding behaviors with the high silt sediment. However, Co had a poor correlation with the other six heavy metals in various binding phases, except with organic matter. Binding behaviors of Pb in the phases of bound to carbonates and exchangeable were different from the other six heavy metals. Cu was inversly correlated with the other six heavy metals in its binding behavior with reducible phases (Fe-/Mn-oxides). PMID- 11381781 TI - Effects of groundwater velocity on sampling intervals for contaminant-detection networks in aquifers. AB - This study evaluated how groundwater velocity affects the sampling interval of a groundwater monitoring network and its ability to intercept contaminant plumes before reaching a buffer zone boundary. A computer simulation model tested the detection capability of a groundwater-monitoring network in different groundwater velocity settings. A 0.4-m/d velocity registered a maximum sampling interval of four months. By comparison, decreasing the velocity to 0.04 m/d resulted in a 3 year maximum sampling interval. Groundwater velocity exerts a strong control on the maximum sampling interval of contaminant detection networks in aquifers. Assigning an arbitrary sampling interval, that does not take into account a site's hydrogeology, could waste financial resources or allow contaminants to reach a buffer zone boundary without being detected. PMID- 11381780 TI - Activated sludge immobilization using the PVA-alginate-borate method. AB - The PVA-alginate-borate method was used successfully to immobilize activated sludge. The optimum polyvinyl alcohol (PVA) concentration in the immobilized sludge was determined to be 10-12.5%. A minimal alginate of 1% in the beads was needed to prevent bead agglomeration. If the pH of the saturated boric acid was adjusted to 7.0 prior to allowing droplets of the mixture of PVA, alginate and sludge to enter the solution, a high level of sludge activity could be maintained in the beads formed. During the continuous operation of a fluidized bed reactor, with hydraulic retention time (HRT) in the range of 24-3 h (BOD loading from 0.176 to 0.766 kg/d/m3) and an aeration rate at 1.0 L/min, more than 90% of BOD5, NH4(+)-N and TKN were removed. The immobilized sludge exhibited satisfactory mechanical stability without apparent breakage. PMID- 11381782 TI - Weibull modeling of the Fenton's oxidation process. AB - A modified version of the two-parameter Weibull survival curve is used to investigate its applicability to analyze the influence of the Fenton's Reagent on the decomposition of 2,4-dinitrophenol solutions (DNP). Fenton's Reagent is one type of advanced oxidation processes (AOPs) in which strong oxidants, hydroxyl radicals, are generated. Fenton's Reagent was maintained at a ratio of 0.5 mM H2O2 to 0.1 mM Fe2+ at the start of all experiments. By varying the initial DNP concentration, we are not only able to correlate parameters (a) and (b) defined in the modified curve to the Fenton's oxidation process, but also interpret their physical significance on the process. The scale parameter (a) can be used to illustrate the rate of the decomposition of DNP in solutions. The imbalance between decomposition rate and initial DNP concentration can be easily observed with the aid of parameter (a). By defining a specific decomposition rate function, the shape parameter (b) indicates the strength of the oxidation power provided by the Fenton's Reagent. When (b) > 1, the existence of a maximum specific decomposition rate shows a sufficient supply of the hydroxyl radicals, whereas a first-order decomposition of DNP is obtained when (b) = 1. A lack of oxidation power becomes obvious if (b) < 1, showing a high dose of initial DNP in a solution. Knowing the physical interpretation of both parameters, they are further applied to design the Fenton's oxidation process. PMID- 11381783 TI - Treatment of highly polluted groundwater by novel iron removal process. AB - The removal of ferrous iron (Fe(II)) in groundwater has been generally achieved by simple aeration, or the addition of an oxidizing agent. Aeration has been shown to be very efficient in insolubilization ferrous iron at a pH level greater than 6.5. In this study, pH was maintained over 6.5 using limestone granules under constant aeration to oxidize ferrous iron in groundwater in a limestone packed column. A sedimentation unit coupled with a membrane filtration was also developed to precipitate and filtrate the oxidized ferric compound simultaneously. Several bench-scale studies, including the effects of the limestone granule sizes, amounts and hydraulic retention time on iron removal in the limestone packed column were investigated. It was found that 550 g/L of the 7 8 mesh size limestone granules, and 20 min of hydraulic retention time in the limestone packed column, were necessary for the sufficient oxidation of 40 mg/L of iron(II) in groundwater. Long-term operation was successfully achieved in contaminated waters by removing the iron deposits on the surface of the limestone granule by continuous aeration from the bottom of the column. Periodic reverse flow helped to remove caking and fouling of membrane surface caused by the continuous filtration. Recycling of the treated water from the membrane right after reverse flow operation made possible an admissible limit of iron concentration of the treated water for drinking. The pilot-scale process was constructed and has been tested in the rural area of Korea. PMID- 11381784 TI - Modification of coagulation and Fenton oxidation processes for cost-effective leachate treatment. AB - Physicochemical processes have been used to remove nonbiodegradable organic compounds in leachate generated from sanitary landfills. In this study, a coagulation process combined with Fenton oxidation was evaluated for the removal of refractory organics in leachate. Recycling of sludge generated from Fenton oxidation to a coagulation process was attempted to reduce operation cost and to improve organic removal rate. The addition mode of Fenton reagent was also modified to obtain a better organic removal rate and more cost-effective operation. With Fenton sludge recycling, 9% higher COD removal was obtained, and the sludge to be disposed could be reduced up to 50%. Also, the coagulant could be reduced by 50%. For Fenton oxidation process, stepwise addition of reagents gave a 5% higher COD removal, and a 25% reduction in chemical consumption. PMID- 11381785 TI - Accumulation of metals in vegetation from an alkaline artificial soil. AB - This research was conducted to determine whether elevated concentrations of metals in biosolids would result in increased accumulation of these metals in plants growing on an alkaline, artificial soil created by addition of municipal sewage biosolids to inorganic waste materials during reclamation of a filled lime settling basin. Accumulation of metals in vegetation growing on this alkaline, artificial soil was compared with accumulation in vegetation growing on a natural, reference soil that had not been amended by biosolids. Although the concentrations of Cd, Cr, Hg, Ni, Ag, and Zn were greater in the artificial soil than in the reference soil, only Cd was found at greater concentrations in vegetation growing on the artificial soil. These greater concentrations were only observed in wheat grass (Agropyron sp.), and not in orchard grass (Dactylis glomerata L.). Also, the accumulation ratio, which is the ratio of the metal concentration in plant tissue to that in the soil, for wheat grass was much lower than other published values. There were no differences in the concentrations of Ba and Pb in the two soils, but the concentrations of these two metals in orchard grass growing on the reference soil were greater than that in either grass on the artificial soil. The differences in plant uptake of metals appear to result from differences in soil pH between the artificial and reference soils. Many metals are generally more soluble and bioavailable at the lower pH of the reference soil (pH = 5.4-6.6) than at the higher pH of the artificial soil (pH = 7.8-8.2). As a result, transfer of metals to vegetation growing on the alkaline, artificial soil is less than that in vegetation growing on the reference soil despite the greater concentrations of metals in the artificial soil. PMID- 11381786 TI - The effect of hydration on adsorption and desorption of heavy metals in soils. AB - Cr, Ni, and Cd adsorption-desorption on five characterized Taiwan soils was studied. The potential for toxicity and the fate of metals in the soils is dependent upon the ability of the metals to desorb into the aqueous phase. To simulate field conditions, the soils were subjected to wet-dry cycles. The amount and rate of desorption was found to decrease with increasing number of cycles. The wet-dry cycle effect in the desorption of metals from soils is related to the soil composition, with desorption being easiest from sandy soil with low organic content. The desorption process is much slower than adsorption process. More than 95% of each metal adsorption takes place within one hour, and the amount of metal adsorbed from solution reached equilibrium in one day. In contrast, less than 20% metal desorption could be attained after 3 days each of four wet-dry cycles for Cr, Ni, and Cd. PMID- 11381787 TI - Application of path analysis to urinary findings of cadmium-induced renal dysfunction. AB - In order to identify some causal relations among various urinary indices of cadmium-induced renal dysfunction, such as glucose, total protein, amino nitrogen, beta 2-microglobulin (beta 2-m), metallothionein (MT), and cadmium (Cd), we applied path analysis method to previous epidemiological studies targeting the residents of the Cd-polluted Kakehashi River basin of Ishikawa Prefecture, Japan. We obtained a diagram-termed path model, representing some causal relations among the above urinary indices. It shows that urinary Cd is located at the beginning point in the diagram, and Cd-induced renal dysfunction develops in the following order: Cd exposure-->increase of beta 2-m and/or MT excretion-->increase of amino-N and/or total protein excretion-->increase of glucose excretion. It was proved mathematically, that in the case of both males and females, increased excretions of beta 2-m and/or MT were the most sensitive urinary indices of the early stage of chronic Cd-induced renal dysfunction. PMID- 11381788 TI - Solubility of polynuclear aromatic hydrocarbons in supercritical carbon dioxide. AB - New data for the solubility of five three- to five-ring polynuclear aromatic hydrocarbons, anthracene, phenanthrene, triphenylene, chrysene and perylene, in supercritical carbon dioxide are reported. Measurements were obtained with a recirculating high-pressure system employing on-line monitoring to verify equilibrium. The influence of water and the role of methanol as a solubility enhancer were investigated. Heats of Solution and Heats of Mixing were calculated from van't Hoff plots. Van't Hoff methodology also served to demonstrate the consistency of the data. PMID- 11381789 TI - Cost of treating heart failure in an Irish teaching hospital. AB - BACKGROUND: The prevalence of heart failure is 3 to 20 per 1,000 population, but may exceed 100 per 1,000 in the over 65 age group. Some 1-2% of the total healthcare budget is consumed in the management of heart failure. AIM: As hospital costs account for approximately 70% of this expenditure we determined the cost of treating heart failure in an Irish teaching hospital. METHODS: Cost evaluation was from the hospital perspective using a microcosting detailed collection of resources used. RESULTS: The average cost of a hospital admission for cardiac failure was IR 2,146 Pounds. The average cost per day was IR 193 Pounds. Approximately 75% of hospital costs were associated with ward costs. Medications accounted for 3.5% of total costs. CONCLUSION: The availability of Irish cost data is essential for the assessment of the cost-effectiveness of therapeutic interventions for the treatment of heart failure in our healthcare system. PMID- 11381790 TI - A & E services in Ireland: the potential role of general practice in accident and emergency services. AB - BACKGROUND: In 1996, Irish accident and emergency (A&E) departments had approximately 1.2 million visits. General practitioners (GPs) have been shown to work efficiently in A&E. AIM: This study aimed to describe the current A&E structures in Ireland and the potential contribution of general practice. METHOD: Questionnaires were sent to all 43 Irish A&E departments seeking information on staffing levels, training posts and interest in the role of GPs within the department. RESULTS: Thirty-four (79%) hospitals responded, representing at least 71% of all A&E visits. Eleven (32%) had A&E consultants. In 16 (47%) hospitals the A&E department was supervised by other consultants; in 14 supervision was for five hours per week or less. Seven hospitals had no consultant supervision. Twenty-six (76%) had NCHDs assigned to the department. Only 11% of NCHDs were in training in A&E medicine. Six departments employed GPs but 28 said they would like to do so. Most wished GPs to see non-urgent cases but one-third wished them to see all cases. Current staffing levels had little relationship with departmental workload. CONCLUSIONS: The limited consultant supervision and small numbers of NCHDs in training for A&E medicine raise concerns about staffing. Most hospitals want GPs to work in their A&E departments. This has implications for training and for the interface between general practice and the A & E department. PMID- 11381791 TI - Prevalence and predictors of recurrent IgA nephropathy following renal transplantation. AB - BACKGROUND: IgA nephropathy is a common glomerulonephritis for which there is no effective cure. It may recur after renal transplantation and cause graft loss. AIMS: To determine the prevalence and predict recurrence of IgA disease in transplant recipients. METHODS: A retrospective analysis was performed of all renal allografts in patients with IgA disease attending a National Renal Unit between 1984 and 1995. An immunopathological grading system was devised to assess the severity of disease at initial presentation and each patient was assigned a simple severity index. RESULTS: A total of 42 patients with IgA disease received 44 renal allografts. Biopsies were performed in 21 of the 44 transplants. Recurrence was diagnosed in five grafts (24%) and recurred only in the 'moderate' and 'severe' IgA groups. Recurrence was associated with younger age, glomerular crescents on the original renal biopsy, better donor/recipient HLA matching and greater number of rejections. CONCLUSION: The prevalence of recurrent IgA disease following transplantation in an Irish population is less than that reported at other centres (24% versus 60%). The severity of the original disease and transplant factors may predict recurrence post-transplantation. PMID- 11381793 TI - Genetic archaeology and the origins of the Irish population. PMID- 11381794 TI - Irish medical journals. PMID- 11381792 TI - Prevalence of risk factors for ischaemic stroke and their treatment among a cohort of stroke patients in Dublin. AB - BACKGROUND: The majority of strokes are due to ischaemia. Risk factors include atrial fibrillation, hypertension and smoking. The incidence can be reduced by addressing these risk factors. This study examines the prevalence of risk factors and their treatment in a cohort of patients with ischaemic stroke registered on a Dublin stroke database. METHODS: Patients admitted to any of three acute hospitals with a diagnosis of stroke during a one-year period in 1997/98 were registered on a database using the European Stroke Database format. Data relating to common risk factors were analysed. RESULTS: There were 238 ischaemic stroke cases registered. The most frequent medical risk factors were: hypertension (45%), atrial fibrillation (27.3%), and previous disabling or non-disabling stroke (33.2%). There was an increasing trend with advancing age for atrial fibrillation (p < 0.001). Some 23% (54/233) were current smokers. A significantly higher proportion of patients with no medical risk factors were smokers or consumed excessive alcohol compared with those who had medical risk factors. CONCLUSION: Medical risk factors for stroke were common among stroke patients and not optimally treated, particularly with regard to atrial fibrillation and previous stroke. Smoking was a major behavioural risk factor among younger patients and much health gain could be achieved in this group through primary prevention strategies. PMID- 11381795 TI - Autoimmune keratolysis. PMID- 11381796 TI - [Congenital rubella syndrome in infants treated at a pediatrics hospital]. AB - Congenital rubella syndrome (CRS) has been considered an uncommon problem in Mexico. OBJECTIVE: To analyze and describe clinical features of CRS cases in infants from a pediatric hospital in Mexico City during an 8 year period. MATERIAL AND METHODS: DESIGN: retrospective study. Patients younger than 18 months of age with a positive serologic test for IgG and IgM rubella antibodies were included. Antibodies were measured by an immunoenzymatic microparticles assay. RESULTS: Fifty-six cases were identified, 42 complete clinical records were available for review. Of these, 23 (54.7%) were female and 19 (45.3%) male. Median for age was five months. A total of 9/42 mothers (21%) had history of rash during pregnancy. IgM antibodies were detected in 15 infants and IgG in 27. Major manifestations were ocular (74%), neurologic (66%), and congenital heart disease (67%). Congenital cataracts were detected in 69%, in 52% hepatomegaly, in 43% jaundice, in 40% anemia, in 48% thrombocytopenia, and hearing loss 19%. CONCLUSION: Five confirmed cases with CRS per year in one hospital indicate a high frequency. Only a small percentage of women had a history of rash during pregnancy. CRS must be investigated in infants with ocular, neurologic, and congenital heart diseases. PMID- 11381797 TI - [Study of 50 patients with sporotrichosis. Clinical and laboratory assessment]. AB - Because sporotrichosis is the most frequent subcutaneous mycosis in Mexico and the clinical aspect is not always characteristic, the aim of this study was to evaluate laboratory diagnosis techniques. Fifty patients with clinical diagnosis of subcutaneous sporotrichosis were studied including clinical and epidemiologic data. Metabolic antigen was used to elicit delayed hypersensitivity skin reaction in all patients. Exudate was plated on Sabouraud agar and biopsy material was submitted to indirect immunofluorescence and histopathology. Results showed that sporotrichosis frequency was higher in women (62%), in children and adolescents under 20 years of age (34%) and adults older than 50 years of age (28%). Disease was predominant in farmers (44%) followed by housewives (30%). Lymphangitic form accounted for 82% of cases and these were localized in upper limbs (54%). In 66% of cases, histopathology showed S. schenckii yeasts; hypersensitivity skin reaction was positive in 76% and culture in 94%. By indirect immunofluorescence, parasitic elements were demonstrated in all patients corresponding to both sensitivity and specificity 100%. In this work, indirect immunofluorescence was the most efficient sporotrichosis diagnostic method followed by culture, hypersensitivity skin reaction, and histopathologic study. PMID- 11381798 TI - [Surgical treatment of giant mediastinal tumors]. AB - A total of 1814 cases of thoracic neoplastic pathology were registered during a period of 15 years (1984-1998), among which mediastinal tumors represented 7.38% (134 cases). There were 14 cases of giant mediastinal tumors, invading more than two thirds of the hemithorax and more than one of mediastinal compartment. Teratomas were the most common in this group. Surgical excision is indicated most cases. The results of surgical treatment on the basis of clinical, histologic and imagenologic studies are presented. The blind dissection of the adherences of the tumor to anatomic intrathoracic structures is a troublesome step; other surgical difficulties are commented upon. Eleven tumors were successfully excised, only one patient died. The surgical treatment of giant mediastinal tumors is a true challenge for the thoracic surgeon and the pulmonologist. PMID- 11381799 TI - [Comment on "Surgical treatment of giant mediastinal tumors"]. PMID- 11381800 TI - [Systemic inflammatory response syndrome: new concepts]. AB - Systemic inflammatory response syndrome is secondary to several insults whose purpose is to limit and reverse the injury. The outcome and intensity of the inflammatory response is determined by injury extension and balance between inflammatory and compensatory antiinflammatory responses. The interaction of the several soluble and cellular mediators of inflammatory and antiinflammatory responses determine the next evolutive phases: a) local inflammatory response; b) systemic inflammatory response; c) massive systemic inflammatory response; d) immunologic paralysis; and e) immune dissonance. If the last three phases do not control, cell damage can be amplified and thus perpetuating the infectious process and leading the patient to multiple organ dysfunction. Gamma interferon and steroids as modulators of the inflammatory response, in stages of paralysis and immune dissonance has been studied in clinical and experimental trials with promising results, but more studies to are needed validate their usefulness. PMID- 11381801 TI - [Intracellular signals involved in glucose control]. AB - Many proteins are involved in glucose control. The first step for glucose uptake is insulin receptor-binding. Stimulation of the insulin receptor results in rapid autophosphorylation and conformational changes in the beta chain and the subsequent phosphorylation of the insulin receptor substrate. This results in the docking of several SH2 domain proteins, including PI 3-kinase and other adapters. The final event is glucose transporter (GLUT) translocation to the cell surface. GLUT is in the cytosol but after insulin stimulation, several proteins are activated either in the GLUT vesicles or in the inner membrane. The role of the cytoskeleton is not well known, but it apparently participates in membrane fusion and vesicle mobilization. After glucose uptake, several hexokines metabolize the glucose to generate energy, convert the glucose in glycogen and store it. Type 2 diabetes is characterized by high glucose levels and insulin resistance. The insulin receptor is diminished on the cell surface membrane, tyrosine phosphorylation is decreased, serine and threonine phosphorylation is augmented. Apparently, the main problem with GLUT protein is in its translocation to the cell surface. At present, we know the role of many proteins involved in glucose control. However, we do not understand the significance of insulin resistance at the molecular level with type 2 diabetes. PMID- 11381802 TI - [Neuroendocrine pancreatic tumor]. PMID- 11381803 TI - [Corneal topography in refractive (Lasik) surgery]. PMID- 11381804 TI - [Laryngeal plasmacytoma]. AB - Solitary plasmocytoma is a rare presentation of plasma cell dyscrasia. About 1% od the patients can present with a extramedullary plasmocytoma; although bone presentation is the most frequent, in some cases, soft tissue can be affected. Radiotherapy remain as the election treatment. A case of primary plasmocytoma of the larynx is presented. The patient is alive after radiotherapy. Discussion about the clinical differentiation and also a review of the literature is present. PMID- 11381805 TI - [Dr. Francisco Marin y Moran (1837-1905). Retrospective about an academician scholar from Puebla. Historical research]. PMID- 11381806 TI - [The uncertain future of general surgery. A specialty in the process of extinction]. AB - General surgery was the first specialty of medicine. Nowadays, it is fragmented a progressive number of specialties, and has lost many of its fields. What will be the future of general surgery? What will be its repercussion on teaching surgery? From the point of view of a professor of surgery, our specialty today has nothing like what it was in the middle of the past century. Every day, its field of action becomes more and more reduced. Probably in the future, training in surgery will be limited to 2 years of basic knowledge, before the resident goes into the specialty of his/her choice. PMID- 11381807 TI - [Vertebral osteomyelitis associated with epidural block]. AB - Infectious complications after epidural anesthesia are infrequent and the most common are epidural and subdural abscess. We report one rare case of vertebral osteomyelitus associated with an epidural catheter and review the literature. PMID- 11381808 TI - [A new susceptibility gene in Alzheimer's disease]. PMID- 11381809 TI - [Menopause: are selective estrogen modulators (SERMs) an alternative treatment to hormonal replacement?]. PMID- 11381810 TI - [Cuff pressure in endotracheal intubation: should it be routinely measured?]. AB - OBJECTIVE: To determine the relationship between endotracheal tube cuff pressure and post-surgical tracheal pain. MATERIAL AND METHODS: Cross-sectional study of forty subjects who required endotracheal intubation for elective surgery allocated into two groups according to tube cuff pressure. Subjects who had cuff pressure equal to or less than 42 Mmhg were assigned to Group A, and those with cuff pressure higher than 42 Mmhg to group B. Cuff pressure measurements were carried out previously to removing the endotracheal tube. Low-pressure, high volume type of tube was used in all subjects. Tracheal pain was evaluated at 60 min and 24 h after extubation. RESULTS: There were not differences in the intubation time required (117 +/- 36.9 min vs. 133 +/- 64.9 min, p = 0.3) or in the number of tracheal tubes used in both groups. Tracheal pain was similar in both groups 60 min after extubation, but at 24 h persisted only in 10% of subjects in group A and 53.3% of B, p = 0.02. The correlation between tracheal pain and tube cuff pressure at 24 h was 0.76, p = 0.00001. CONCLUSIONS: High tube cuff pressure is a related factor to the tracheal pain so must be considered a routine monitoring of cuff pressure and device to avoid cuff pressure that exceeds the necessary minimum. PMID- 11381811 TI - [Inaugural speech at the 87th academic year. National Academy of Medicine]. PMID- 11381812 TI - [Neurodegenerative diseases leading to dementia. Published in GMM No. 6, year 2000, Vol 136]. PMID- 11381813 TI - [Bacteremia and pseudobacteremia caused by coagulase-negative Staphylococcus in children]. AB - OBJECTIVE: To compare patients with Coagulase Negative Staphylococcus bacteremia (CoNS-B) and pseudobacteremia (CoNS-PB) in a pediatric hospital. METHODS: Descriptive and comparative study between children diagnosed with CoNS-B and CoNS PB. RESULTS: A total of 159 children with CoNS positive blood cultures were evaluated. 66 children were classified as CoNS-B (41.5%) and 93 as CoNS-PE3 (58.5%). On the average CoNS was isolated on day 21 among children with bacteremia (B) and on day 2 in those with PB (p < 0.01). Excluding newborns, patients with B and PB had on average 2.6 and 1.1 positive cultures respectively. Most children with bacteremia were at the intensive care unit (67.2%), while patients with PB were mostly detected at the emergency room. Using logistic regression analysis, we found four factors independently associated with CoNS bacteremia: total parenteral nutrition (OR 5.4; 95% CI 2.2-12.9), low birth weight (OR 2.6; 95% CI 1.1-5.9), catheters placed by cut-down technique (OR 1.9; 95% CI 1.1-3.8), and inmuno-compromised patients (OR 2.7; 95% CI 1.1-6.7). Resistance to oxacilin was reported in 71.7% of the CoNS isolated. The overall mortality associated to CoNS-B was 6%. Among children with CoNS-PB, 10% received antibiotics, half of them vancomycin. CONCLUSIONS: These results show that CoNS-B occurs mainly as a nosocomial episode. CoNS-PB more likely resulted from specimen contamination at collection, being responsible for almost 60% of all positive blood cultures. The false-positive results caused unnecessary administration of antibiotics in a significant proportion of CoNS-PB events and have a potential impact upon the emergence of resistant pathogens. PMID- 11381814 TI - Hispanics, Latinos, or Americanos: the evolution of identity. AB - This essay identifies and categorizes terms used to designate the Hispanic/Latino population in the United States. It provides an analysis framing the process of ethnic self-designation within an ethnopolitical and psychosocial context. The analysis concludes by presenting mestizaje and transculturation as processes involved in the evolution of Latino identity. PMID- 11381815 TI - What's missing from multicultural competency research: review, introspection, and recommendations. AB - Much of the multicultural counseling research has pointed out the need for counselors to become culturally competent to appropriately address the needs of an emerging diverse community. To date, however, this body of research has largely focused on counselor competencies, with little attention being given to the client perspective of multicultural counseling competencies. This article discusses the importance of integrating the client perspective within the historical context of the multicultural literature by examining client preferences and expectations, as well as the adequacy of the current empirical data. The authors also raise the issue of politics inherent in multiculturalism and discuss how this affects the research. A number of recommendations are made as a guide for future research. PMID- 11381816 TI - Relating ethnic identity, acculturation, and attitudes toward treating minority clients. AB - This study assessed the relationship among the ethnic identity, acculturation, and self-concepts of minority health care professionals and their attitudes toward treating minority clients and using alternative therapies. The sample consisted of 150 medical students of Indian descent. Strong ethnic identity and acculturation related to high self-esteem, self-clarity, the likelihood of treating more minority patients, and the use of alternative therapies. The more "Westernized" participants expressed less of a preference for serving minority patients and believed themselves to be less effective in serving minorities. Gender differences in some aspects of ethnic identity and attitudes were found. The results imply that a focus on acculturation and ethnicity in all client clinician psychological research is critical, and especially in cases of cultural matching. PMID- 11381817 TI - Equivalencies regarding the measurement and constructs of self-esteem and major life events in an Asian/Pacific islander sample. AB - Construct, scalar, and functional measurement equivalencies of the Rosenberg Self Esteem Scale (RSES) and Major Life Events checklist (MLE) and the constructs assessed were investigated across groups differentiated on Hawaiian/part-Hawaiian and non-Hawaiian (e.g., Caucasian, Filipino, Hispanic, Japanese, and mixed/2 or more) ethnicity and gender. Initial results from maximum likelihood factoring with promax rotation showed that RSES negatively worded Item 5 loaded with the positively worded Items 1, 2, 4, 6, and 7 on 1 of 2 factors for Hawaiian/part Hawaiian female participants. Similarly, negatively worded Item 8 and the same positively worded items comprised 1 of 2 factors for non-Hawaiian male participants. For the other 2 Ethnicity x Gender groups, factors were respectively comprised of the 5 positively and 5 negatively worded RSES items. Construct equivalence or simple (2-factor) structure underlying the RSES was indicated across the 4 groups after Items 5 and 8 were excluded from a subsequent factoring procedure. Simple structure showed that Factor 1 comprised the positively worded Items 1, 2, 4, 6, and 7, and the remaining negatively worded Items 3, 9, and 10 loaded on Factor 2. Scalar equivalence of the self-esteem and major life events measures was supported by the statistical nonsignificance of the Major Life Events x Ethnicity x Gender interaction effect in multiple regression models. The consistency in the absolute size and direction of the intercorrelations between overall self-esteem, self-esteem Factors 1 and 2, and major life events variables indicated the functional equivalence of respective measures and constructs assessed. Measurement equivalency findings concerning the RSES and MLE, the constructs measured, and their utility versus caution against their use in multiethnic studies were discussed. PMID- 11381818 TI - Assessing sexual harassment among Latinas: development of an instrument. AB - This study draws from multiple data sources and analytic approaches to offer preliminary evidence on how to assess sexual harassment among Latinas working in the United States, particularly working-class Mexican American women with moderately low acculturation. First, focus group data were collected from 45 Latinas to identify culture-specific manifestations of sexually harassing behavior. These data informed the development of new items for the Sexual Experiences Questionnaire (SEQ: L. F. Fitzgerald et al., 1988), making it more appropriate for administration to Latinas with limited education. A 2nd Latina sample (N = 476) then completed this and other scales in a paper-and-pencil survey. Complete-link hierarchical cluster analyses of the SEQ data, based on a random half-sample of these women, revealed an underlying 3-factor structure. Confirmatory factor analyses on the 2nd half-sample confirmed that this factor model fit the data well, and both the individual factors and the larger scale appear highly reliable. Implications of both new and discarded SEQ-Latina items, the 3-factor structure, and relationships among the factors were discussed. PMID- 11381819 TI - Black identity in biracial black/white people: a comparison of Jacqueline who refuses to be exclusively black and Adolphus who wishes he were. AB - Two biracial college freshmen, both of whom identify as Black, were chosen from a larger sample of participants in a qualitative study of biracial identity development to exemplify the differences in the paths that 2 biracial individuals could take to achieve racial identity resolution. Through the case study method, the authors describe the course and progression of racial identity development (RID) in these 2 individuals and discuss some key themes in their lives that have contributed to the development of their RID. The purposes are fourfold: to describe nonclinical subjective experiences of being biracial in the United States, to explore the differences in the paths that 2 biracial individuals can take to achieve what looks superficially like similar Black racial identity resolution, to demonstrate how identifying as Black can have different meanings and consequences for 2 biracial people, and to contribute to the differentiation of Black RID from biracial Black/White RID. The authors raise questions about the generalizability of monoracial Black and ethnic identity theories to biracial individuals. PMID- 11381820 TI - Management of hypertension: do we know all the questions? PMID- 11381821 TI - Anatomical variations of internal jugular vein as seen by "Site Rite II" ultrasound machine--an initial experience in Pakistani population. AB - OBJECTIVE: To determine the anatomical variations of the internal jugular vein (IJV) in Pakistani adult population with the help of Site Rite II ultrasound machine. MATERIAL AND METHOD: The right IJV relation to the carotid artery was visualized at four different landmarks (angle of the mandible, thyroid cartilage, cricoid cartilage, and the supraclavicular area). Size of IJV in comparison to carotid artery was also seen. RESULTS: In 49 cases the IJV was found in aberrant relation to carotid artery at the angle of the mandible (p value < 0.05), 22 at the thyroid cartilage, 20 at the cricoid cartilage, and 46 at the supraclavicular area (p value < 0.05). In 93% of cases the IJV was found to be larger than the carotid artery. CONCLUSION: Care should be taken while cannulating IJV at the angle of the mandible and supra clavicular area by external landmark guided technique. Ultrasound guided technique should be used in every anticipated difficult case. PMID- 11381822 TI - Maternal deaths in a developing country: a study from the Aga Khan University Hospital, Karachi, Pakistan 1988-1999. AB - OBJECTIVE: The maternal deaths occurring over a twelve-year period (1988-1999) in a tertiary referral center were reviewed. The purpose of the study was to assess the causes of these maternal deaths. SETTING: The Aga Khan University Hospital (AKUH) Karachi, Pakistan. METHODS: The medical records of maternal deaths were reviewed. These were women who had either registered for delivery at the hospital; or were referred from another hospital or from home, when an emergency developed. They were either admitted to the Medicine, Surgery and the Obstetrics and Gynaecology Departments at the hospital. RESULTS: A total of 81 maternal deaths were identified, of which five were the registered patients. Causes of deaths were eclampsia, puerperal sepsis and pulmonary embolism. The maternal mortality ratio in the registered patients was 20 per 100,000 live births. Ninety percent of the women were between the age group of 15-35 years. Of these forty two percent were primigravidas, forty four percent of the women died due to direct causes, of which sepsis was the most common cause and accounted for twenty five percent of the total deaths. Indirect causes were responsible for 55.6% of the deaths, including hepatic failure in 21%, other infectious disease in 17% and malignancy in 5% of the cases. CONCLUSION: In developing countries other than obstetrical causes, infectious diseases contribute to the death of women during childbearing years. Comprehensive medical services and adequate obstetrical emergency services can lower maternal mortality rates at all levels. PMID- 11381824 TI - Bronchodilator, spasmolytic and calcium antagonist activities of Nigella sativa seeds (Kalonji): a traditional herbal product with multiple medicinal uses. AB - OBJECTIVE: The seeds of Nigella sativa locally known as "Kalonji" has been used in traditional medicine for the treatment of a variety of diseases including diarrhoea and asthma. The crude extract of N. sativa seeds (Ns.Cr) was studied in vitro for its possible spasmolytic and bronchodilator activities to rationalize the folkloric uses. METHODS: Isolated rabbit jejunum and guinea-pig tracheal preparations were set up in Tyrode's and Kreb's solutions respectively and aerated with 5% CO2 in oxygen. Isotonic and isometric responses were measured on Bioscience oscillograph and Grass polygraph respectively. RESULTS: The Ns.Cr caused a dose-dependent (0.1-3.0 mg/ml) relaxation of spontaneous contractions in rabbit jejunum. Ns.Cr also inhibited K(+)-induced contractions in a similar dose range, suggestive of calcium channel blockade (CCB). This effect was confirmed when pretreatment of the tissue with Ns.Cr, produced a dose-dependent shift in the Ca++ dose-response curves to the right similar to that of verapamil, a standard calcium channel blocker. In guinea-pig trachea, it caused relaxation of carbachol-, histamine- or K(+)-induced contractions indicating CCB. Activity directed fractionation revealed that the CCB activity is concentrated in the petroleum ether fraction, which was found to be approximately 10 times more potent than the crude extract both in jejunum and tracheal preparations. CONCLUSION: These data indicate that the crude extract of Nigella sativa seeds exhibits spasmolytic and bronchodilator activities mediated possibly through calcium channel blockade and this activity is concentrated in the organic fraction. Its usefulness for diarrhoea and asthma in traditional medicine, appears thus to be based on a sound mechanistic background. PMID- 11381823 TI - Milk composition changes--a simple and non-invasive method of detecting ovulation in lactating women. AB - OBJECTIVE: To assess the possibility of detecting ovulation through observing changes in composition of milk and using this as a simple and non-invasive method. METHOD: Twenty five normal lactating women (menstruating and non menstruating), varying in age from 18-35 years and taking no hormonal contraceptives were included in this study. Hormonal estimation was done by radioimmunoassay and changes in milk composition were observed by measuring the concentration of Na+, Cl-, K+ and lactose. RESULTS: Changes were observed in milk composition of four menstruating women (ovulatory), characterized by significant increase in concentration of Na+ and Cl- and decrease in concentration of lactose and K+. However, in one non-menstruating woman who was found to have ovulation, these findings could not be observed. CONCLUSION: Results show that the determination of Na+, K+, Cl- and lactose in milk of lactating women have a high value for detection of ovulation. PMID- 11381825 TI - A survival analysis of metastatic breast cancer in Pakistani patients. AB - OBJECTIVE: To determine the overall survival of metastatic breast cancer in the Pakistani patients and compare it with published information. METHOD: The design was a retrospective analysis of metastatic breast cancer patients from breast cancer database. A total of 137 patient based information was available for review and analysis. RESULTS: An overall median survival of 2.83 years was noted in metastatic breast cancer patients. CONCLUSION: This survival figure in this study compares favorably to those published in the literature. PMID- 11381826 TI - Lack of correlation between alpha-fetoprotein and tumor size in hepatocellular carcinoma. AB - OBJECTIVE: To determine whether there was a correlation between tumor size and alpha feto-protein (alpha-FP) levels in hepatocellular carcinoma. SETTING: Retrospective study in tertiary referral center with specialist Oncology services in Southern Pakistan. SUBJECTS: Consecutive patients with biopsy-proven hepatocellular carcinoma diagnosed between January 1994 and June 1998. MAIN OUTCOME MEASURES: Correlation between alpha-FP levels and maximum tumor diameter. RESULTS: The mean tumor size was 8.3 +/- 4.2 cm. The mean alpha-FP level was 17,027 ng/ml. Twenty four percent patients had an alpha-FP level which was within the normal limits (< 10 ng/ml). There was no correlation between tumor size and alpha-FP levels (r = -0.155; p = 0.129). CONCLUSION: There was no correlation between the tumor size and alpha-FP levels. PMID- 11381827 TI - Leptin's implications in reproduction: current understandings. PMID- 11381828 TI - Trisomy 9: a case report. PMID- 11381829 TI - The relationship of socio-demographic factors with iron deficiency anaemia in children of 1-2 years of age. AB - OBJECTIVE: To investigate associations between different socio-demographic factors with iron deficiency anaemia in Pakistani children of 1-2 years of age. METHODOLOGY: A case control study, with 50 cases and 100 controls, was conducted at the Community Health Centre, an outpatient clinic of the Aga Khan University, Karachi, Pakistan, between July 1993-July 1995. Informed consent was taken from mothers for their children's blood tests and a questionnaire was administered to them. The data was analyzed using chi-square, t-tests and logistic regression. RESULTS: The numbers of pregnancies, live births and living children were more among cases than controls but the differences were not statistically significant. Although father's education did not show a significant association (OR 1.35, 95% CI 0.22-8.33), maternal education was significantly associated with the children's anaemic status (OR 3.55, 95% CI 1.40-9.02). The difference in monthly incomes between families of cases and controls was the most significant variable among all those studied (p-value 0.006). CONCLUSION: This study showed that while lack of maternal education and low monthly family incomes are both significantly associated with the development of childhood anaemia, low monthly income is most significant. PMID- 11381830 TI - Flow cytometric analysis of childhood leukemias. AB - OBJECTIVE: To collect demographic data for childhood (less than 15 years) leukemias in Karachi, describe the accuracy of the cell surface markers routinely used in the flow cytometric analysis of leukemic cells and arrive at an ideal panel of antibodies for analyzing leukemic samples. MATERIALS AND METHODS: Data from 62 consecutive cases of childhood leukemias referred to the Department of Pathology, Aga Khan University Hospital, (AKUH) between January 1995 and December 1998 was analyzed using Epi Info Version 6. Flow cytometry on all samples was performed using standard protocols. RESULTS: The mean age of patients was 8.2 years and 49 (79%) were males. Fifty (81%) had acute lymphoblastic leukemias of which 50% were CD10 positive and 24% CD10 negative Pre-B cell leukemias. Among all Pre B cell All 98% were positive for CD19, 96% for CD22, 89% for HLA-DR and 67% for CD10. Of the 10 AML cases, 100% were positive for CD33, 90% for CD13, 80% for CD19 and 70% for HLA-DR. CONCLUSION: The mean age in this study population was significantly higher and percentage of CD10 positive Pre-B All is lower than that in the West. Both these factors might be responsible for the poorer prognosis of these patients. It is not possible to specify a minimum or maximum panel of antibodies that should be used for phenotyping all cases of childhood leukemias. A certain degree or redundancy is essential in any panel of antibodies used for flow cytometry of leukemias. PMID- 11381831 TI - Culture and systems of thought: holistic versus analytic cognition. AB - The authors find East Asians to be holistic, attending to the entire field and assigning causality to it, making relatively little use of categories and formal logic, and relying on "dialectical" reasoning, whereas Westerners are more analytic, paying attention primarily to the object and the categories to which it belongs and using rules, including formal logic, to understand its behavior. The 2 types of cognitive processes are embedded in different naive metaphysical systems and tacit epistemologies. The authors speculate that the origin of these differences is traceable to markedly different social systems. The theory and the evidence presented call into question long-held assumptions about basic cognitive processes and even about the appropriateness of the process-content distinction. PMID- 11381832 TI - Conjunctive representations in learning and memory: principles of cortical and hippocampal function. AB - The authors present a theoretical framework for understanding the roles of the hippocampus and neocortex in learning and memory. This framework incorporates a theme found in many theories of hippocampal function: that the hippocampus is responsible for developing conjunctive representations binding together stimulus elements into a unitary representation that can later be recalled from partial input cues. This idea is contradicted by the fact that hippocampally lesioned rats can learn nonlinear discrimination problems that require conjunctive representations. The authors' framework accommodates this finding by establishing a principled division of labor, where the cortex is responsible for slow learning that integrates over multiple experiences to extract generalities whereas the hippocampus performs rapid learning of the arbitrary contents of individual experiences. This framework suggests that tasks involving rapid, incidental conjunctive learning are better tests of hippocampal function. The authors implement this framework in a computational neural network model and show that it can account for a wide range of data in animal learning. PMID- 11381833 TI - Heritability estimates versus large environmental effects: the IQ paradox resolved. AB - Some argue that the high heritability of IQ renders purely environmental explanations for large IQ differences between groups implausible. Yet, large environmentally induced IQ gains between generations suggest an important role for environment in shaping IQ. The authors present a formal model of the process determining IQ in which people's IQs are affected by both environment and genes, but in which their environments are matched to their IQs. The authors show how such a model allows very large effects for environment, even incorporating the highest estimates of heritability. Besides resolving the paradox, the authors show that the model can account for a number of other phenomena, some of which are anomalous when viewed from the standard perspective. PMID- 11381834 TI - Multialternative decision field theory: a dynamic connectionist model of decision making. AB - The authors interpret decision field theory (J. R. Busemeyer & J. T. Townsend, 1993) as a connectionist network and extend it to accommodate multialternative preferential choice situations. This article shows that the classic weighted additive utility model (see R. L. Keeney & H. Raiffa, 1976) and the classic Thurstone preferential choice model (see L. L. Thurstone, 1959) are special cases of this new multialternative decision field theory (MDFT), which also can emulate the search process of the popular elimination by aspects (EBA) model (see A. Tversky, 1969). The new theory is unique in its ability to explain several central empirical results found in the multialternative preference literature with a common set of principles. These empirical results include the similarity effect, the attraction effect, and the compromise effect, and the complex interactions among these three effects. The dynamic nature of the model also implies strong testable predictions concerning the moderating effect of time pressure on these three effects. PMID- 11381835 TI - Executive control of visual attention in dual-task situations. AB - A theory of executive control is presented that proposes that executive processes control subordinate processes by manipulating their parameters, reconfiguring them to respond in accord with the current task set. It adopts C. Bundesen's (1990) theory of visual attention (TVA) and R. M. Nosofsky and T. J. Palmeri's (1997) exemplar-based random walk (EBRW) as the theory of subordinate processes. It assumes that a task set is a set of TVA and EBRW parameters sufficient to perform a task and that set switching involves changing those parameters. The theory solves 2 computational problems that emerge in dual-task situations: the binding problem and the serial order problem. It can perform dual tasks in series or in parallel but prefers the serial strategy because it is faster and it solves the binding problem naturally. The theory accounts for concurrence cost, set switching cost, crosstalk between tasks, and the modulation of crosstalk by task set. PMID- 11381836 TI - Traps and gaps in action explanation: theoretical problems of a psychology of human action. AB - This article deals with the question of whether human action can be explained empirically by a psychological theory that refers to intentions, expectancies, and evaluations as determinants. In contrast with the majority of action theories in psychology and philosophy, a logical connection between action and intention is defended and, consequently, a causal relationship between action and intention is refuted. This is illustrated by reference to one of the most widely known and applied psychological action theories: the theory of planned behavior (I. Ajzen, 1991). However, the logical-connection argument can be circumvented if the existing research findings are reinterpreted as part of a psychology of intention. This article demonstrates the value of such an approach for future research. However, the final section of the article outlines some further fundamental theoretical difficulties for this perspective. PMID- 11381837 TI - A context noise model of episodic word recognition. AB - Item noise models of recognition assert that interference at retrieval is generated by the words from the study list. Context noise models of recognition assert that interference at retrieval is generated by the contexts in which the test word has appeared. The authors introduce the bind cue decide model of episodic memory, a Bayesian context noise model, and demonstrate how it can account for data from the item noise and dual-processing approaches to recognition memory. From the item noise perspective, list strength and list length effects, the mirror effect for word frequency and concreteness, and the effects of the similarity of other words in a list are considered. From the dual processing perspective, process dissociation data on the effects of length, temporal separation of lists, strength, and diagnosticity of context are examined. The authors conclude that the context noise approach to recognition is a viable alternative to existing approaches. PMID- 11381838 TI - [Impact of weekly administration of folic acid on folic acid blood levels]. AB - OBJECTIVE: To evaluate the effect of weekly administration of 5 mg. folic acid (FA) intake on folic acid blood levels. MATERIAL AND METHODS: This concurrent comparative study was conducted in 1998, in urban and rural areas of Nuevo Leon State, Mexico. The study population consisted of 74 women who delivered a child during 1997, 39 of whom had a child with a neural tube defect. Women were given 5 mg. of folic acid, weekly for 3 months. Blood levels of folic acid were determined by radioimmunoassay (RIA) at baseline, and a week after taking the last folic acid dose. Data are presented as means and standard deviations. RESULTS: Ninety per cent of the women had significantly increased folic acid intraerythrocytary blood levels. Red cell folate increased from 150.49 +/- 31.17 ng/ml to 184.21 +/- 35.53 ng/ml (p < 0.005). Plasma folate increased from 5.93 +/ 1.98 ng/ml a 7.03 +/- 2.5 ng/ml (p < 0.05). Eighty-two per cent of women reached intracellular levels above 160 ng/ml. CONCLUSIONS: The weekly administration of folic acid is a simple and efficient method that could be used to supplement this vitamin to low-income populations. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 11381839 TI - Effect of the quality of health care on permanent inability secondary to femoral fractures due to occupational accidents. AB - OBJECTIVE: To construct and validate an indicator for evaluating the quality of care for femoral fractures, and to assess the contribution of the quality of health care as a determinant of partial permanent inability secondary to femoral fractures due to occupational accidents. MATERIALS AND METHODS: The study was conducted from January to December 1995 at Mexican Institute of Social Security. The instrument was designed with experts' contribution along different stages and validated using implicit criteria and factorial analysis. A case-control study was then conducted to evaluate the contribution of the quality of care to inability secondary to femoral fractures. Cases were 108 active workers with permanent inability secondary to femoral fracture; controls were 94 active workers with fracture of femur but no permanent inability. Logistic regression modeling was used to establish the association between quality of care and partial permanent inability, adjusting by relevant variables. RESULTS: The ultimate indicator of quality of care consisted of the following: Timely care, presurgical management, surgical management, and fracture complications. A final score over 229 points meant that the worker had received good quality of care. Workers getting 229 or less points had received poor quality of care. Forty-eight (44%) cases and 66 (70%) controls received good quality of medical care. The likelihood of partial permanent inability was almost three times higher among workers given poor quality of care (OR 2.95; 95% CI 1.5-5.5). According to the multivariate model, predictors of partial permanent inability were: Having exposed or epiphysiary fractures, being re-submitted to surgery, having less than 90 days of rehabilitation care, and receiving deficient medical care. CONCLUSIONS: The constructed instrument was validated. The level of the quality of care received by workers is a determining factor for the generation of partial permanent inability. In workers having femoral fractures, it is important to consider timely medical care and early rehabilitation, to reduce the high incidence and prevalence of this medical problem in Mexico. The English version of this paper is available too at: http://www.insp.mx/salud/index.html. PMID- 11381840 TI - [Reproducibility of spirometry in Mexican workers and international reference values]. AB - OBJECTIVE: To describe spirometric function and adjustment to foreign prediction equations in Mexican workers claiming work related disability. MATERIAL AND METHODS: We reviewed 5771 spirometries done at the Mexican National Institute of Respiratory Diseases performed with equipment and methods proposed by the American Thoracic Society. With the spirometries we generated multiple regression equations separated for men and women based on age and height, compared to other in common use reported by Knudson and Hankinson in North America and by Quanjer in europeans. RESULTS: 80% of the tests were reproducible for FVC and FEV1 according to ATS, whereas 10% were reproducible for neither. Mean FVC in men was 12% above values reported by Quanjer, 22% above Knudson, 3% above Hankinson and 6% above Rodriguez-Reynaga, whereas similar values for women were 18%, 10%, 0% and 1%. Excluding obese and those who had less than 2 acceptable maneuvers, the numbers increase by 1-2%. FEV1 was also above predicted. CONCLUSIONS: Most workers requesting disability are able to generate a reproducible spirometry. However for the same gender, age and height, workers had a FEV1 and a FVC above normal values reported by Knudson and Quanjer and are more similar to those reported by Hankinson in Mexican-Americans. While a set of appropriate reference values are obtained, regression equations obtained from the studied group will generate less error in the evaluation of disability in mexican workers. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 11381841 TI - [Incidence of Helicobacter pylori infection in a cohort of infants in the State of Morelos]. AB - OBJECTIVE: To study the incidence of Helicobacter pylori infection in infants from the State of Morelos, Mexico. MATERIAL AND METHODS: A cohort of 110 healthy infants was studied between 1997 and 1999. Serum samples were collected from mothers and their infants at 2, 6, 18, and 24 months of life. All serum samples were tested for antibodies against Helicobacter pylori with the ELISA test. A questionnaire was used to collect socio-economic and clinical data. Associations among selected variables and Helicobacter pylori infection were determined using Fisher's exact test. RESULTS: Two thirds of mothers and six (5.5%) infants tested positive. Two of the six positive infants were born to positive mothers; both of them became negative before age two. The other four infants remained positive. Although not statistically significant, a vaginal birth and more than five people living in the household are possible risk factors for infant H. pylori infection. CONCLUSIONS: Despite the previously reported high prevalence of infection by H. pylori in Mexican children, in this population we found a low incidence of infection in infants up to two years of age. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 11381842 TI - [Fluoride intake and excretion in children of Hermosillo, Sonora, Mexico]. AB - OBJECTIVE: To estimate the fluoride intake and urinary excretion among 8-9 year old school children. MATERIALS AND METHODS: A cross-sectional study was conducted from February to November 1997. Study subjects were 31 male children from Hermosillo, Mexico, living in three neighborhoods with different drinking water fluoride levels. Fluoride intake was measured through water intake and collection of food samples ingested in 24 hr. Fluoride excretion was measured in 24-hour urine samples. The fluoride concentration was measured by the potentiometer method using a fluoride-specific ion electrode. RESULTS: Statistically significant differences of fluoride levels in drinking water were found in the 3 different neighborhoods: 2.77, 0.78 and 0.54 mg/L, respectively (p < 0.01). The daily mean intake of fluoride for the neighborhood with the highest fluoride level was 5.41 mg/d, while mean intakes for the other two neighborhoods were 2.31 and 1.51 mg/d, respectively. Thirty-two percent of the children had fluoride intake above the recommendation values for this age group; sixty per cent of these children lived in the neighborhood with the highest fluoride level. Drinking water accounted for 63% of total fluoride intake, and the mean water intake was 1.8 liters. The mean excretion of fluoride was 1.61 mg/24 hours (range 0.18 to 8.1 mg/24 hr). Stepwise multiple regression showed an association between drinking water fluoride and fluoride urinary excretion (R2 = 0.56 p < 0.001). CONCLUSIONS: Fluoride intake in the neighborhood with the highest fluoride level was twice the daily recommended value for 8-9 year-old children. Drinking water was the main source of fluoride intake. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 11381843 TI - [Case-control epidemiological studies: theoretical bases, variants and applications]. PMID- 11381845 TI - [Questioning the pertinence of including variables related to climatic changes and daily activities]. PMID- 11381844 TI - [Genetic diseases of the mitochondrial DNA in humans]. AB - Mitochondrial diseases are a group of disorders produced by defects in the oxidative phosphorylation system (Oxphos system), the final pathway of the mitochondrial energetic metabolism, resulting in a deficiency of the biosynthesis of ATP. Part of the polypeptide subunits involved in the Oxphos system are codified by the mitochondrial DNA. In the last years, mutations in this genetic system have been described and associated to well defined clinical syndromes. The clinical features of these disorders are very heterogeneous affecting, in most cases, to different organs and tissues and their correct diagnosis require precise clinical, morphological, biochemical and genetic data. The peculiar genetic characteristics of the mitochondrial DNA (maternal inheritance, polyplasmia and mitotic segregation) give to these disorders very distinctive properties. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 11381846 TI - [Expectancies, alcohol drinking and associated problems in university students in Mexico City]. AB - OBJECTIVE: To assess the relationship between alcohol use expectancies, drinking patterns, and alcohol-related problems, among college students in Mexico City. MATERIAL AND METHODS: This cross-sectional study was conducted in October 1998. Study subjects were 678 male and female college students aged between 17 and 25 years, from private and public schools. Alcohol expectancies were measured through the self-reported "Alcohol Expectancy Questionnaire" (AEQ), adapted for this population, with a high overall internal reliability coefficient (alpha = 0.93). RESULTS: Thirty-one per cent of study subjects, mainly males, reported heavy drinking (5 drinks or more per drinking occasion in the previous year), while 17% were non-consumers. Relationships for each of the AEQ scales, drinking patterns, and alcohol-related problems, were analyzed through a structural equations model. Analysis of variance showed a statistically significantly association between six AEQ subscales--alcohol as a facilitator of a group interaction, verbal expression, disinhibition, sexual enhancement, reduction of psychological tension and arousal aggression-(F = 5.23, df = 1, p < 0.05). CONCLUSIONS: Study findings from other countries show that effects attributed to alcohol, in anticipation of the drinking episode (expectancies), are closely related to alcohol intake. Our results showed that heavy drinkers had higher scores in six AEQ subscales, particularly those who reported having alcohol related problems in the previous year. Alcohol expectancies related to higher alcohol intake were: facilitation of social interaction, decreasing psychological tension, and increasing arousal/aggression. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 11381847 TI - [Factors associated with acceptance of postpartum tubal ligation among HIV infected women]. AB - OBJECTIVE: To identify the factors associated with the acceptance of tubal ligation after childbirth among HIV-infected patients. MATERIAL AND METHODS: A case-control study was conducted from March 1988 to February 1999, at Mexico's National Institute of Perinatology (INPer), in 72 HIV-positive pregnant women. Cases were 49 women who accepted postpartum tubal ligation after childbirth, and controls were 23 women who refused this birth control method. Data collected for each patient were demographic characteristics, sexual and reproductive history, and HIV status. Statistical analysis consisted of descriptive measures, Chi 2 or Fisher's exact test for categorical variables, and Student's t test for continuous variables. Odds ratios (OR) with 95% CI were used to compare groups and potential confounders were assessed by stratified analysis with the Mantel Haenszel method. RESULTS: The patients' mean age was 25.5 +/- 5.5 years. The median gestation period was 27 weeks (range 7 to 40 weeks); 16 women (22.2%) had no prenatal care visits at INPer. The median time of HIV positivist awareness was 9 months (range 1 to 108). Variables associated with acceptance of tubal ligation were: having a prior childbirth (OR 11.1, 95% CI 3.4 to 36), pregnancy care from 1995 onward (OR 4.7, 95% CI 1.7 to 13.3), and having given birth to an HIV infected child (OR 4.6, 95% CI 1.05 to 23.1). Stratified analysis showed no modification of the strength of association of these variables with acceptance of tubal ligation. CONCLUSIONS: A prior childbirth was the most important predictor of tubal ligation acceptance. The English version of this paper is available at: http://www.insp.mx/salud/index.html. PMID- 11381848 TI - Stemming the tide. PMID- 11381849 TI - Botanical briefs: the mango tree--Mangifera indica L. PMID- 11381850 TI - Lentigo. AB - A lentigo is a small pigmented macule with a sharply circumscribed border. There are multiple clinical and etiologic forms. Lentigines are often initially identified shortly after birth, although they may appear later in childhood. Certain varieties are associated with systemic abnormalities. Histologic findings include epidermal hyperplasia with increased pigmentation of the basal layer. PMID- 11381851 TI - Photo quiz: pigmentation due to minocycline. PMID- 11381852 TI - Cutaneous Acanthamoeba in a patient with AIDS: a case study with a review of new therapy; quiz 386. AB - GOAL: To describe the presenting signs of an Acanthamoeba infection. OBJECTIVES: Upon completion of this activity, dermatologists and general practitioners should be able to: 1. Discuss the clinical presentation of Acanthamoeba infection. 2. Describe the conditions that make a patient susceptible to Acanthamoeba. 3. Outline treatment options for Acanthamoeba infection. CME: This article has been peer reviewed and approved by Michael Fisher, MD, Professor of Medicine, Albert Einstein College of Medicine. REVIEW DATE: April 2001. This activity has been planned and implemented in accordance with the Essentials and Standards of the Accreditation Council for Continuing Medical Education through the joint sponsorship of Albert Einstein College of Medicine and Quadrant HealthCom, Inc. The Albert Einstein College of Medicine is accredited by the ACCME to provide continuing medical education for physicians. Albert Einstein College of Medicine designates this educational activity for a maximum of 1.0 hour in category 1 credit toward the AMA Physician's Recognition Award. Each physician should claim only those hours of credit that he/she actually spent in the educational activity. This activity has been planned and produced in accordance with ACCME Essentials. PMID- 11381853 TI - Erythema elevatum diutinum mimicking extensive keloids; quiz 386. AB - GOAL: To describe a case of erythema elevatum diutinum (EED) that clinically mimicked extensive keloids. OBJECTIVES: Upon completion of this activity, dermatologists and general practitioners should be able to: 1. Describe the clinical presentation and lesion appearance in EED. 2. Discuss the electron microscopic and laboratory findings of EED. 3. Outline treatment options for EED. CME: This article has been peer reviewed and approved by Michael Fisher, MD, Professor of Medicine, Albert Einstein College of Medicine. REVIEW DATE: April 2001. This activity has been planned and implemented in accordance with the Essentials and Standards of the Accreditation Council for Continuing Medical Education through the joint sponsorship of Albert Einstein College of Medicine and Quadrant HealthCom, Inc. The Albert Einstein College of Medicine is accredited by the ACCME to provide continuing medical education for physicians. Albert Einstein College of Medicine designates this educational activity for a maximum of 1.0 hour in category 1 credit toward the AMA Physician's Recognition Award. Each physician should claim only those hours of credit that he/she actually spent in the educational activity. This activity has been planned and produced in accordance with ACCME Essentials. PMID- 11381854 TI - Treatment of lentigo maligna. AB - Lentigo maligna (LM) is an indolent form of melanoma in situ with the potential to progress to invasive melanoma. Early detection and adequate treatment prior to development to invasive melanoma are essential. Definitive excision with negative margins is currently the treatment of choice for LM. Conventional excision, Mohs micrographic surgical excision, and nonexcisional methods of treatment of LM will be discussed. PMID- 11381855 TI - Update on malignant melanoma in children. AB - Malignant melanoma is a rare event in children. Yet, the overall incidence has consistently risen in the past 20 years. Thus, the likelihood that our pediatric patients will develop malignant melanoma is increasing. Previously, the bulk of lesions were estimated to occur in children with large congenital melanocytic nevi. Recent reports, however, have highlighted new risk factors for malignant melanoma in children, while demystifying other entities previously believed to have a grave prognosis. Knowledge of risk factors and participation in public health efforts toward prevention and early intervention can help the practitioner protect pediatric patients from this malignancy. PMID- 11381856 TI - Water-slide alopecia. AB - A 29-year-old male presented with large, symmetric, alopecic patches on the posterolateral aspects of both calves. A detailed history revealed that the individual had recently attended a water-slide amusement park. Repeated frictional trauma between the legs and the slide resulted in these alopecic patches. Friction, especially when encountered during sports-related and recreational activities, should be included in the differential diagnosis of well defined alopecic patches. PMID- 11381857 TI - Malignant melanoma in African Americans. AB - Although rare, malignant melanoma (MM) is a real and serious risk for African Americans. African Americans have a proportionately higher incidence of acral melanoma, both the acral lentiginous melanoma (ALM) histologic subtype and subungual melanoma (SM). MM is more likely to be diagnosed at a later stage in African Americans and carries a worse prognosis. Given these facts, the relatively simple and inexpensive primary and secondary preventions for MM should be standard, particularly in the African American patient. PMID- 11381858 TI - Evaluation of pigmented lesions of the nail unit. AB - Acquired pigmentary changes of the nail are secondary to a number of etiologies. These include nail matrix nevi; physical induction secondary to trauma; malignant melanoma; nutritional deficiencies; inflammation secondary to lichen planus; endocrine causes such as Addison's disease; or secondary to bacterial, fungal, or viral infections. The most important task faced by clinicians is to distinguish benign from malignant etiologies of nail pigmentation. We will briefly review the various entities that can yield dyspigmentation and their differentiation from melanoma of the nail. PMID- 11381859 TI - Extrafacial granuloma faciale: report of a case and response to treatment. AB - Granuloma faciale (GF) is a rather uncommon form of chronic vasculitis that infrequently involves extrafacial sites. Treatment of this disease is extremely challenging. We report a case of GF with extrafacial lesions and a unique response to treatment. The diseases that are clinical and histologic mimics of this disorder, as well as a review of various treatment modalities, are discussed. PMID- 11381860 TI - Lichen sclerosus et atrophicus affecting the wrists and left ankle and clinically simulating lichen planus. AB - Lichen sclerosus et atrophicus (LSA) is a disease of unknown etiology, although hereditary, endocrine, and autoimmune factors are known to be involved. Although the anal and genital regions are predominantly affected, 2.5% of patients only present with extragenital lesions--particularly of the trunk, neck, and upper limbs. The wrists, palmoplantar regions, nipples, and face are less commonly involved. The possible relationship between LSA and both lichen planus and localized scleroderma (morphea) has not been clearly established, although in a number of cases, several of these conditions have been found simultaneously. We report the case of a 61-year-old woman with LSA lesions affecting only the wrists and left ankle. The unusual character of this presentation is pointed out, along with its clinical similarity to lichen planus. PMID- 11381862 TI - Cutaneous malignancy in albinism. AB - Albinism is a disorder of hypopigmentation affecting the skin, appendages, and eyes. Ultraviolet light-induced cutaneous tumors are common in patients with albinism due to reduced or absent protection from melanin. Squamous cell carcinoma (SCC) is the number one skin tumor seen in patients who are albinos. Although nonmelanomatous skin cancers are more common in patients with albinism, dysplastic nevus and melanoma present a greater diagnostic challenge in this group because of their hypopigmented appearance. We report 2 cases of cutaneous malignancies in patients who had oculocutaneous albinism (OCA). The first case involves a 45-year-old man with OCA type 2 (OCA2) who developed a large SCC of the neck. The second case involves a 24-year-old man with Hermansky-Pudiak syndrome (HPS) who developed amelanotic melanoma. In both cases, hypopigmentation of the lesions contributed to a delay in diagnosis. We review the clinical, diagnostic, and therapeutic concerns for patients with albinism who have cutaneous malignancies. PMID- 11381861 TI - Linear IgA bullous dermatosis associated with vancomycin and disseminated varicella-zoster infection. AB - Linear IgA bullous dermatosis (LABD) is characterized by linear deposits of IgA at the basement membrane zone. Most cases are idiopathic, but medications, infections, autoimmune disorders, and malignancies have been documented as potential inducers. We report a case where both vancomycin and varicella-zoster infection were present as triggers. PMID- 11381863 TI - Occupational exposures and associated health effects among sanitation landfill employees. PMID- 11381864 TI - Preference reversals in grading systems for retail food facilities. AB - The authors asked 120 university students about their willingness to dine in restaurants if those restaurants had received inspection grades of A, B, or C under three different scenarios: 1) the restaurant is one they normally dine in, 2) the restaurant has been closed for health violations but recently reopened, and 3) the restaurant has been given a score (88, 78, 68, or 58) but no grade. Using a paired-samples sign test, the authors discovered differences among the scenarios with statistical significance above 99.9 percent. Subjects were less willing to dine if a restaurant had been recently closed and were more willing to dine if only scores were given. The results indicate three cases of preference reversal from the normal order of A, B, and C: 1) subjects preferred a grade of B over A if the B restaurant had a score of 88 and the A restaurant had recently been closed; 2) subjects preferred a grade of C over B if the C restaurant had a score of 78 and the B restaurant had recently been closed; and 3) subjects preferred a score of 68 (which would earn closure in many jurisdictions) over a legitimate grade of C at a recently closed restaurant. In addition, subjects preferred a score of 88 to a grade of B and a score of 78 to a grade of C, even though those scores and respective grades are assumed to be equivalent. The study population was limited to university students, who may not necessarily represent the broader population. Similar cognitive effects have, however, been documented throughout the literature on risk perceptions. In any event, the results indicate a need for closer examination of grading systems. On the basis of preference reversal principles, the authors propose ways to improve grading systems for retail food facilities and suggest an agenda for further research. PMID- 11381865 TI - Levels of polychlorinated biphenyls (PCBs) in fish: the influence on local decision making about fish consumption. AB - In 1989, the level of Aroclor 1254 (a mixture of polychlorinated biphenyls with 42 percent chlorine by weight) in fish collected from a recreational pond in Toledo, Ohio, was reported to be 44.4 milligrams per kilogram (mg/kg), prompting local health officials to declare a "no fishing advisory" for the pond. A second study conducted in 1990 did not identify elevated levels of polychlorinated biphenyls (PCBs) in fish tissue. In other words, the two studies gave conflicting results. The purpose of this study was to determine the level of Aroclor in fish tissues and then evaluate whether consumption of fish from the pond would pose a serious health risk. Fish samples collected on several occasions in 1998 and 1999 were filleted, and tissues were analyzed, as composite or individual samples, with gas chromatography. The levels of Aroclor 1254 ranged from 0.2 mg/kg in white crappies to 1.0 mg/kg in carp. These levels, while far less than the level reported in 1989, nevertheless were greater than 0.05 mg/kg, which is the maximum level established by the Great Lakes Fish Advisory Task Force for "no restriction in fish consumption." Levels of other Aroclor formulations ranged from less than the 0.02 mg/kg (the minimum detectable limit) to 0.1 mg/kg. Given current knowledge about the potential health consequence of exposure to PCBs and the results of this study, the authors have recommended that local health officials develop a new fish consumption guideline for PCBs. Any decisions about maintaining or lifting the restrictions on the pond, however, should be based on additional studies that determine the levels of other chemicals that are present in the pond and deemed hazardous to human health. PMID- 11381866 TI - The rights of regulators. PMID- 11381867 TI - Readable writing by scientists and researchers. PMID- 11381868 TI - Achieving compliance with the radium standards for drinking water in a midwestern community: a case study. AB - The problem of elevated levels of radium in the drinking-water supply of DeKalb, Illinois, a Midwestern community about 65 miles west of Chicago with approximately 35,000 residents, has been a contentious issue for over a decade. The central players in the controversy include a group of concerned citizens, city officials, the Illinois Pollution Control Board, the Illinois Environmental Protection Agency, and the U.S. Environmental Protection Agency (U.S. EPA). Achieving a satisfactory resolution to the problem has been a long, drawn-out process that illustrates how ill-timed proposals, changing risk assessments, different perceptions of risk, and the high costs of compliance can influence the direction of risk management decisions. The purpose of this study is to analyze how these factors sustained the debate and prevented an expeditious solution to the problem. The study uses document analysis as its primary research tool. The questions raised by the controversy, along with their implications for environmental policy, are discussed, as are some of the more important lessons learned from the case. Because of many uncertainties, there were no clear winners in the controversy, although, in the context of the most recent U.S. EPA risk assessments, the community is now poised for better protection from the potential dangers of radium in drinking water than it might have been had citizen action been absent. PMID- 11381869 TI - Marine eutrophication: the need for a new indicator system. PMID- 11381870 TI - Predicting effects of toxic chemicals in the marine environment. AB - Ecological risks are typically characterized in risk assessment procedures by considering the ratio between exposure concentrations and critical effect concentrations. In OECD countries, critical effect concentrations are typically derived from laboratory-based ecotoxicity tests using well-defined protocols on a limited number of species. More and more countries in the tropics are adopting this approach in environmental assessment, protection, and management. In this article we consider a number of issues associated with such an approach, and in particular potential problems with extrapolating effects on individuals observed in laboratory-based ecotoxicological investigations to effects on ecosystems. It is hoped that by making explicit some of the assumptions made in the potential limitations of these tests, we can better target our limited resources to protect valuable and vulnerable systems. PMID- 11381871 TI - Arsenic, antimony, selenium and other trace elements in sediments of the La Paz Lagoon, peninsula of Baja California, Mexico. PMID- 11381872 TI - Contamination by persistent organochlorines and butyltin compounds in the west coast of Sri Lanka. AB - Concentrations of polychlorinated biphenyls (PCBs), dichlorodiphenyl trichloroethane and its metabolites (DDTs), isomers of hexachlorocyclohexane (HCHs), hexachlorobenzene (HCB), chlordane compounds (CHLs) and butyltin compounds (mono-, di- and tributyltin) were determined in biota and sediments collected from locations on the west coast of Sri Lanka. The PCB concentrations found in fish from the Kelani River mouth were higher than those from other Asian developing countries. The pattern of organochlorine accumulation was in the order of PCBs > DDTs > CHLs > HCHs > HCB, and resembled those reported for most developed countries. The accumulation pattern of butyltins in fish was in the order of TBT > DBT > MBT suggesting recent exposure to TBT sources. Total butyltin concentrations found in rabbit fish liver (11-38 micrograms/g wet wt), collected from the Colombo dockyard, could be the highest ever reported for fish in the world. PMID- 11381873 TI - Baseline metal concentrations in coastal Labrador sediments. AB - Levels of Cr, Cu, Fe, Pb, Ni, and Zn in surface sediment from the Voisey's Bay area of coastal Labrador showed no evidence of recent anthropogenic input of metals. Metal concentrations in surface sediments, normalized to Li, fell within the 95% confidence limits of the background levels. Further analysis showed that the Li-metal regression lines from the surface sediments and sediments from 30-cm depth had the same slope and intercepts, suggesting that there was no difference in the metal content of the sediments at the two depths. Li-metal relationships can be used as a measure of the natural variability of the metal concentrations for the region and will serve as a baseline against which future anthropogenic affects may be assessed. PMID- 11381874 TI - Changes in the marine environment of Port Kembla Harbour, NSW, Australia, 1975 1995: a review. AB - Data on water quality, sediment quality and aquatic organisms in Port Kembla Harbour from the 1970s to the 1990s are reviewed. In the 1970s, the marine environment of Port Kembla Harbour was in poor condition as a result of pollution from heavy industries. Elevated concentrations of pollutants were found in water, sediment and fish in the harbour; aquatic biodiversity was limited and many fish kills were reported. With the implementation of pollution reduction programs (required by legislation changes) by the industries surrounding the harbour since the 1970s, pollution in the harbour has been reduced dramatically, and the quality of the marine environment of the harbour has noticeably improved. Large reductions in the concentrations of certain toxic wastes and heavy metals in water have occurred. Marine life has returned to the whole harbour (parts were described in 1977 as abiotic). Contaminants in fish have decreased. Despite this achievement, however, there is still considerable room for improvement in the quality of the marine environment of the harbour. PMID- 11381875 TI - Recovery of the macrozoobenthic community after severe dystrophic crises in a Mediterranean coastal Lagoon (Orbetello, Italy). AB - The Orbetello lagoon, one of the largest Western Mediterranean lagoons, was affected by high algal blooms and severe anoxic crises in 1992 and 1993, with fish mortality throughout most of the lagoon and a drastic reduction of benthic species. Many measures were undertaken between 1994 and 1996 to remove the severe eutrophication of this lagoon. Such measures included harvesting of the floating seaweed biomass, removal of all wastewater and sea channel enlargement by dredging activity. The aim of the present research was to study the macrozoobenthic assemblage in 1999 and to compare it with previous research in order to assess the recovery of the lagoon. A total of 106 taxa and 45,175 individuals were collected in three areas (sea inlets, western and eastern lagoon) having different organic matter loading during winter and summer. The results of multivariate and correlation analyses indicated that trophic status and its seasonal dynamics were crucial in determining species distribution among the different areas. Moreover, comparison of the macrozoobenthic structure between winter 1995 and 1999 showed differences in species composition and structural parameters in the western and eastern lagoon, where a clear recovery of benthic assemblages was observed in 1999. However, the dominance of opportunistic species in most of the lagoon and the decrease in some structural parameters during summer suggested that this basin still presented signs of disturbance in 1999, six years after the last extended anoxic crises and even after completion of the restoration measures carried out in the environment. It was hypothesized that a secondary disturbance, sustained by a high seasonal release of nutrients from the sediments, could create considerable deviations from the expected improvement in benthic conditions. PMID- 11381876 TI - Chemical forms of Pb, Zn and Cu in the sediment profiles of the Pearl River Estuary. AB - The chemical forms of heavy metals (Pb, Zn and Cu) in sediment cores of the Pearl River Estuary were studied using a sequential chemical extraction method. The isotope ratios of 206Pb/207Pb in various chemical fractions were also measured to assess the potential Pb sources. Zinc and Cu were mainly associated with the residual fraction. The Fe-Mn oxide and organic/sulphide fractions were the next important phases for Zn and Cu, respectively. For Pb, different chemical partitioning patterns were found among different sediment cores. Most Pb was associated with the residual fraction in the sediments. In some sediment profiles, the major phase of Pb in the top layers was the Fe-Mn oxide fraction. The proportion of Pb in the Fe-Mn oxide fraction decreased significantly with increasing depth. Among the different depths, the 206Pb/207Pb isotope ratios in the residual fraction remained fairly stable, with a mean value of 1.202, which may represent the natural background value. The 206Pb/207Pb ratios in the exchangeable fraction were the lowest among the five fractions, particularly in top sediments, showing the anthropogenic inputs of heavy metals from recent rapid industrial development in the surrounding region. For the other three non residual fractions, there was a similar trend of increasing 206Pb/207Pb ratios down the profile. Results from this study are useful in assessing both the chemical changes for heavy metals in marine sediments and the potential of heavy metal release into the water environment of an estuary area. PMID- 11381877 TI - Trace metals in fish and invertebrates of three California coastal Wetlands. AB - Metal concentrations were measured in selected fish and invertebrate species from Mugu Lagoon, Malibu Lagoon and Ballona Wetlands in southern California in order to assess the extent of metal contamination in these three wetlands. Ranges of element concentrations (in microgram/g) found in biota were: Zn 12-650; Cu 1.9 440; Ni < 1-37; Cr < 1-55; Pb < 0.5-6.8; As < 1-8.5; Se < 1-3.8; Cd < 0.2-0.90; and Ag < 0.3-5.9. Relative to previous studies of California biota, the highest metal concentrations found were for chromium and nickel. The highest levels were in one of the two bottom-dwelling fish (juvenile Leptocottus armatus) (55 micrograms/g) and the two water-column fish sampled (Fundulus parvipinnis and Atherinops affinis) (30 and 24 micrograms/g). At Ballona Lagoon, elevated levels of copper and silver were found in the bivalve Tagelus californianus (440 and 5.9 micrograms/g). Chromium and nickel appeared to be most persistent in fish from Mugu (4.6-55 and 2.6-37 micrograms/g), the most northern site and an active military base, and Ballona (< 1-30 and < 1-16 micrograms/g), believed to be the most metal-contaminated site. Compared to previously measured metal concentrations in species of California coastal waters, these regions revealed higher levels of chromium, nickel, silver, arsenic, zinc, copper and, to a lesser extent, cadmium and selenium. Chromium and silver were present at high enough levels at all three sites to be considered environmental health hazards. PMID- 11381878 TI - Photosynthetic pigments and peroxidase activity as indicators of heavy metal stress in the Grey mangrove, Avicennia marina (Forsk.) Vierh. AB - Mangroves have been observed to possess a tolerance to high levels of heavy metals, yet accumulated metals may induce subcellular biochemical changes, which can impact on processes at the organism level. Six month-old seedlings of the grey mangrove, Avicennia marina (Forsk.) Vierh, were exposed to a range of Cu (0 800 micrograms/g), Pb (0-800 micrograms/g) and Zn (0-1000 micrograms/g) concentrations in sediments under laboratory conditions, to determine leaf tissue metal accumulation patterns, effects on photosynthetic pigments (chlorophyll a, chlorophyll b and carotenoids), and the activity of the antioxidant enzyme peroxidase. Limited Cu uptake to leaves was observed at low sediment Cu levels, with saturation and visible toxicity to Cu at sediment levels greater than 400 micrograms/g. Leaf Pb concentrations remained low over a range of Pb sediment concentrations, up to 400 micrograms/g Pb, above which it appeared that unrestricted transport of Pb occurred, although no visible signs of Pb toxicity were observed. Zn was accumulated linearly with sediment zinc concentration, and visible toxicity occurring at the highest concentration, 1000 micrograms/g Zn. Significant increases in peroxidase activity and decreases in photopigments were found with Cu and Zn at concentrations lower than those inducing visible toxicity. Significant increases in peroxidase activity only, were found when plants were exposed to Pb. Positive linear relationships between peroxidase activity and leaf tissue metal concentrations were found for all metals. Significant linear decreases in photosynthetic pigments with increasing leaf tissue metal concentrations were observed with Cu and Zn only. Photosynthetic pigments and peroxidase activity may be applicable as sensitive biological indicators of Cu and Zn stress, and peroxidase activity for Pb stress in A. marina. PMID- 11381879 TI - Composition and distribution of beach debris in Orange County, California. AB - Many studies have quantified debris collected on beaches around the world. Only a few of those studies have been conducted in the United States, and they are largely limited to semi-quantitative efforts performed as part of volunteer clean up activities. This study quantifies the distribution and composition of beach debris by sampling 43 stratified random sites on the Orange County, California coast, from August to September 1998. We estimated that approximately 106 million items, weighing 12 metric tons, occur on Orange County beaches. The most abundant items were pre-production plastic pellets, foamed plastics, and hard plastics. Debris density on the remote rocky shoreline was greater than that on high-use sandy beaches for most debris items. This finding partially reflects the periodic clean-up of high-use beaches by local municipalities, and also indicates that a high percentage of the observed debris was transported to the site from waterborne sources. PMID- 11381880 TI - Persistent organochlorine concentrations in sediment and fish from Atlantic coastal and brackish waters off Savannah, Georgia, USA. PMID- 11381881 TI - Use of a telemetered dispensing system for controlling nutrient additions to experimental patch reefs in the ENCORE study at One Tree Island, Great Barrier Reef, Australia. AB - A telemetrically controlled system was developed to add nutrients automatically to experimental patch reefs in a remote marine environment. The experiment, called ENCORE, was done in the lagoon of One Tree Island, a remote research station at the southern end of the Great Barrier Reef. Nutrient dispensing units (NDUs), moored adjacent to patch reefs in the lagoon, were telemetrically linked to a base station on the island. The base station, about 3 km away from the furthest NDU, consisted of a dedicated computer, controller and radio transmitter, which relayed coded signals to a radio receiver mounted on each NDU. This activated a solenoid valve to discharge a measured quantity of concentrated nutrient solution from a measuring chamber using compressed air from a SCUBA tank. The solution was discharged through 4-8 PVC outlets into the basins of the patch reefs to allow thorough mixing. The base station interrogated each NDU to find out if the operation had been successful and stored the information on disk to provide a daily log of operations. Nutrient samples taken within the patch reefs demonstrated that calculated initial mean concentrations of 2 micrograms-at PO4-P l-1 and 10 micrograms-at NH4-N l-1 were achieved. The system we have developed can be used in many situations where regular perturbations need to be introduced to aquatic ecosystems. It uses state-of-the-art technology, yet all components are commercially readily available and relatively inexpensive. Detailed specifications and drawings are available from the Great Barrier Reef Marine Park Authority. PMID- 11381882 TI - The 1997/1998 mass mortality of corals: effects on fish communities on a Tanzanian coral reef. AB - The abnormally high surface temperatures in the world's oceans during 1997/1998 resulted in widespread coral bleaching and subsequent coral mortality. An experiment was performed to study the effects of this coral mortality as well as the influence of the structural complexity on fish communities on a Tanzanian coral reef. Changes in fish communities were investigated on plots of transplanted corals after 88% of these corals had died. A distinct shift in fish community composition was found, although diversity was not affected. Fish abundance rose by 39% mostly due to an increase in herbivores, which seemed to benefit from enhanced algal growth on the dead corals. Fish abundance, species diversity and community composition were also strongly influenced by the structural complexity provided by the live and dead corals. This suggests that a coral reef can support abundant and diverse fish populations also after the corals have died as long as the reef structure is sustained. PMID- 11381883 TI - DDT, PAH and PCB in sediments from the intertidal zone of the Bohai Sea and the Yellow Sea. AB - Current contents of persistent organic contaminants, polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs) and the congeners (naphthalene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, chrysene, benzo(a)anthracene, benzo(a)pyrene and benzo(e)pyrene), DDTs and metabolites in the intertidal sediments of the Bohai Sea and the Yellow Sea were studied. The mean levels of DDTs, PAHs and PCBs were 177.52, 877.2 and 3.4 ng g-1 dry weight, respectively. The spatial trends of the contaminant residue levels were analysed too. PMID- 11381884 TI - Improvements in the epifauna of the Crouch Estuary (United Kingdom) following a decline in TBT concentrations. AB - In the United Kingdom, the use of TBT-based anti-fouling paints on small vessels was banned in 1987, and a biological study of the Crouch Estuary, a yachting centre on the south-eastern English coastline, was initiated in order to monitor any associated changes. The macrobenthic infauna and epifauna were sampled between 1987 and 1992, and again in 1997. Epifaunal sampling in 1997 showed that an earlier increase in sedentary taxa had been sustained, and also that a large population of the native oyster Ostrea edulis had become established upstream. Multivariate analysis revealed continuance of a trend towards directional change in community composition at inner estuary stations, where TBT concentrations were historically much higher. These observations, along with evidence of established populations of a range of ascidian species in the inner estuary after an initial marked increase in densities, provide circumstantial evidence of an underlying improvement in environmental conditions following the TBT ban in 1987. PMID- 11381885 TI - Evidence for the effects of metal contamination on the intertidal macrobenthic assemblages of the Fal Estuary. AB - The intertidal invertebrate macrofauna of five creeks in the Fal estuarine system, Cornwall, UK, is compared with data from 40 locations in six other estuaries in south-west Britain. Multivariate analysis shows that the community composition in the Fal is distinct from all the other estuaries. The differences are principally due to the absence of two crustacean species, Corophium voluntator and Cyathura carinata, and the high abundance of small opportunistic annelid species. The Fal Estuary is heavily contaminated with heavy metals as a result of mining in the catchment, and the faunistic features are commensurate with what is known of the relative sensitivities of marine invertebrate taxa to pollution. PMID- 11381886 TI - A new approach for detecting and mapping sewage impacts. AB - Increased nitrogen loading has been implicated in eutrophication occurrences worldwide. Much of this loading is attributable to the growing human population along the world's coastlines. A significant component of this nitrogen input is from sewage effluent, and delineation of the distribution and biological impact of sewage-derived nitrogen is becoming increasingly important. Here, we show a technique that identifies the source, extent and fate of biologically available sewage nitrogen in coastal marine ecosystems. This method is based on the uptake of sewage nitrogen by marine plants and subsequent analysis of the sewage signature (elevated delta 15N) in plant tissues. Spatial analysis is used to create maps of delta 15N and establish coefficient of variation estimates of the mapped values. We show elevated delta 15N levels in marine plants near sewage outfalls in Moreton Bay, Australia, a semi-enclosed bay receiving multiple sewage inputs. These maps of sewage nitrogen distribution are being used to direct nutrient reduction strategies in the region and will assist in monitoring the effectiveness of environmental protection measures. PMID- 11381887 TI - Persistent organic pollutants in oysters Crassostrea gigas and sand flathead Platycephalus bassensis from Tasmanian estuarine and coastal waters. PMID- 11381888 TI - The ENCORE experiment. PMID- 11381889 TI - Ospar 98/3: an environmental turning point or a flawed decision? AB - While the problems of over-fishing and eutrophication continue largely unchecked, the Ospar Commission, which exists to improve the quality of European seas, requires governments, the public and the oil companies to find 20 billion Pounds to remove millions of tons of steel to demonstrate its commitment to the mantra that 'the seas are not dustbins'. The decision, taken in 1998, was made despite a paucity of evidence to identify the environmental impact redundant rigs create, despite the absence of scientific support to demonstrate that removal is the best practicable environmental option and in the knowledge that the decision had more to do with a green anti-oil agenda than with improving marine quality. It may be shown in the fullness of time not to be in the best interests of the environment. Moreover, and perhaps more importantly, the manner in which the decision was made is lacking in important respects. PMID- 11381890 TI - ENCORE: the effect of nutrient enrichment on coral reefs. Synthesis of results and conclusions. AB - Coral reef degradation resulting from nutrient enrichment of coastal waters is of increasing global concern. Although effects of nutrients on coral reef organisms have been demonstrated in the laboratory, there is little direct evidence of nutrient effects on coral reef biota in situ. The ENCORE experiment investigated responses of coral reef organisms and processes to controlled additions of dissolved inorganic nitrogen (N) and/or phosphorus (P) on an offshore reef (One Tree Island) at the southern end of the Great Barrier Reef, Australia. A multi disciplinary team assessed a variety of factors focusing on nutrient dynamics and biotic responses. A controlled and replicated experiment was conducted over two years using twelve small patch reefs ponded at low tide by a coral rim. Treatments included three control reefs (no nutrient addition) and three + N reefs (NH4Cl added), three + P reefs (KH2PO4 added), and three + N + P reefs. Nutrients were added as pulses at each low tide (ca twice per day) by remotely operated units. There were two phases of nutrient additions. During the initial, low-loading phase of the experiment nutrient pulses (mean dose = 11.5 microM NH4+; 2.3 microM PO4(-3)) rapidly declined, reaching near-background levels (mean = 0.9 microM NH4+; 0.5 microM PO4(-3)) within 2-3 h. A variety of biotic processes, assessed over a year during this initial nutrient loading phase, were not significantly affected, with the exception of coral reproduction, which was affected in all nutrient treatments. In Acropora longicyathus and A. aspera, fewer successfully developed embryos were formed, and in A. longicyathus fertilization rates and lipid levels decreased. In the second, high-loading, phase of ENCORE an increased nutrient dosage (mean dose = 36.2 microM NH4+; 5.1 microM PO4(-3)) declining to means of 11.3 microM NH4+ and 2.4 microM PO4(-3) at the end of low tide) was used for a further year, and a variety of significant biotic responses occurred. Encrusting algae incorporated virtually none of the added nutrients. Organisms containing endosymbiotic zooxanthellae (corals and giant clams) assimilated dissolved nutrients rapidly and were responsive to added nutrients. Coral mortality, not detected during the initial low-loading phase, became evident with increased nutrient dosage, particularly in Pocillopora damicornis. Nitrogen additions stunted coral growth, and phosphorus additions had a variable effect. Coral calcification rate and linear extension increased in the presence of added phosphorus but skeletal density was reduced, making corals more susceptible to breakage. Settlement of all coral larvae was reduced in nitrogen treatments, yet settlement of larvae from brooded species was enhanced in phosphorus treatments. Recruitment of stomatopods, benthic crustaceans living in coral rubble, was reduced in nitrogen and nitrogen plus phosphorus treatments. Grazing rates and reproductive effort of various fish species were not affected by the nutrient treatments. Microbial nitrogen transformations in sediments were responsive to nutrient loading with nitrogen fixation significantly increased in phosphorus treatments and denitrification increased in all treatments to which nitrogen had been added. Rates of bioerosion and grazing showed no significant effects of added nutrients. ENCORE has shown that reef organisms and processes investigated in situ were impacted by elevated nutrients. Impacts were dependent on dose level, whether nitrogen and/or phosphorus were elevated and were often species-specific. The impacts were generally sub-lethal and subtle and the treated reefs at the end of the experiment were visually similar to control reefs. Rapid nutrient uptake indicates that nutrient concentrations alone are not adequate to assess nutrient condition of reefs. Sensitive and quantifiable biological indicators need to be developed for coral reef ecosystems. The potential bioindicators identified in ENCORE should be tested in future research on coral reef/nutrient interactions. Synergistic and cumulative effects of elevated nutrients and other environmental parameters, comparative studies of intact vs. disturbed reefs, offshore vs. inshore reefs, or the ability of a nutrient-stressed reef to respond to natural disturbances require elucidation. An expanded understanding of coral reef responses to anthropogenic impacts is necessary, particularly regarding the subtle, sub-lethal effects detected in the ENCORE studies. PMID- 11381891 TI - Monitoring of the microbial community of a sequencing batch reactor bioaugmented to improve its phosphorus removal capabilities. AB - The acclimatisation of an activated sludge to enhanced biological phosphorus removal conditions was followed after and without bioaugmentation with a low amount of phosphorus-accumulating sludge. Phosphorus removal yields were monitored by conventional analytical methods and microbial communities evolutions were followed by a finger printing molecular technique (PCR-SSCP). While the benefit of the bioaugmentation seems real at the level of the reactor parameters, bioaugmentation speeded up the installation of good and stable phosphorus removal yield, the establishment of the inoculated microbial community in the bioaugmented reactor is still unclear. Both the bioaugmented and the control microbial communities evolved in a similar way to end up with apparently comparable populations. At the time of the experiment, the results suggest that the microbial community inoculated for the bioaugmentation did not establish in the reactor but compensated for phosphorus accumulation until the acclimatisation of an endogenous microbial community arose. PMID- 11381892 TI - Robust soft sensors for SBR monitoring. AB - In wastewater treatment some process variables may be very difficult to measure directly because of the non-availability or excessive cost of sensors. An alternative is to use "soft sensors" that provide online estimates of these inaccessible variables by calculations based on auxiliary measurable variables. Two such sensors are proposed based on extended Kalman filtering or neural networks that could enable the monitoring of nitrate ion, ammonium ion and carbonaceous matter concentrations during nitrification of wastewater. PMID- 11381893 TI - Dynamic control of a continuous-inflow SBR with time-varying influent loading. AB - The conventional sequential control of Sequencing Batch Reactor (SBR) is designed with fixed time periods for various operation phases. However, both of the flow rates and qualities of influent vary over time, therefore, a big capacity of wastewater equalization unit is required to cope with influent variability. Such an equalization unit increases the total treatment costs of the system, especially in a small-scale wastewater treatment system. Moreover, in using a SBR treating a time-varying influent loading with conventional sequential control, the system performance cannot be optimized. This paper presents the application of on-line ORP and pH monitoring to dynamically control a continuous-inflow SBR with time-varying loading of influent flow rates and water qualities. Experiential results show that the dynamic controlled SBR revealed not only achieved better substrate removal efficiencies, but also reduced treatment costs. PMID- 11381894 TI - The feasible sequential control strategy of treating high strength organic nitrogen wastewater with sequencing batch biofilm reactor. AB - The bio-kinetics and feasible sequential control strategy of treating high strength organic carbon and nitrogen wastewater were investigated by conducting the ABS manufacturing wastewater in a series of Sequencing Batch Biofilm Reactors (SBBRs). The on-line ORP, pH, and DO monitoring parameters were applied to identify the feature-points when ammonification, nitrification, and denitrification ends. The carbonaceous matter removal kinetics in the anaerobic and aerobic reaction stages can be expressed by the Michaelis-Menten equation. High efficiency of organic carbon removal and organic nitrogen ammonification in the anaerobic stage can eliminate the substrate competition and activation inhibition to nitrifying organisms in the following aerobic stage. In the sequencing nitrogen removal processes, the producing time and system ORP values of these feature-points have good function relationships with the influent COD loading rates of SBBR, which can be integrated into a set-point (set-time and set ORP) sequential control strategy of nitrogen removal. The automatic control operation results revealed ORP was one of the major control parameters of the sequencing nitrogen removal process in SBBR system and high overall removal efficiency were obtained. PMID- 11381895 TI - Simple process for nutrient removal from food processing effluents. AB - Food processing effluents often contain high levels of nutrients, particularly N. Conventionally, anaerobic ponds are used to purify these effluents in Australia, giving cost-effective removal of BOD but little nutrient removal. It has become apparent that disposal by irrigation as presently practiced normally exceeds sustainable N application rates, thus reduction of nutrient levels before irrigation is becoming mandatory. Meatworks effluent is often discharged to country town sewers, frequently accounting for 50-75% of the nutrient load. Meatworks effluents contain 1,000-4,000 mg/L BOD, 200-400 mg/L TKN and 20-50 mg/LP. Conventional BNR technology can readily remove nutrients from such effluents, either alone or in combination with anaerobic ponds but sludge handling on such a small scale poses economic problems. Laboratory scale trials showed that both BOD removal from meatworks effluent and sludge disposal could be achieved readily in conventional anaerobic ponds. The pond effluent, together with the town sewage if required, could be treated in a sequencing batch reactor (SBR) designed for nitrification/denitrification service. Optimisation of the anaerobic pond operation was required to ensure production of the minimum BOD:N ratio needed for N removal. This paper will describe the design and commissioning of two plants; a demonstration plant installed at a typical sized meatworks in Gippsland, Victoria, and a full scale plant for treatment of combined domestic sewage and effluent from a large meatworks at Longford, Tasmania. In neither case (for different reasons) has P removal yet been required. It was demonstrated that 98% of BOD and up to 95% N removal may be cheaply and readily achieved in the SBR. Where lagoons are used, levels of N suitable for river discharge can be achieved. P can be readily removed by alum treatment when required. PMID- 11381896 TI - Performance of an SBR-plant for advanced nutrient removal, using septic sludge as a carbon source. AB - The Tjustvik SBR-plant outside Stockholm, Sweden has been in operation for four years. The plant has to meet stringent effluent standards, BOD7 < 10 PPM, total N < 15 PPM and total P < 0.3 PPM. The plant is a typical two reactor SBR-plant, sized for about 15,000 inhabitants. During the first year of operation there were difficulties in meeting the P consent level. The difficulties were linked to a deficit of available organic carbon and a secondary phosphorus release. The problem was solved with the addition of septic sludge, in an amount equivalent to about 10,000 to 15,000 inhabitants with respect to the BOD-load. The altered operation resulted in a very stable and good effluent quality from the plant that has been maintained ever since, giving typical discharge levels as follows: BOD7, < 3 mg/l; Total-P, < 0.15 mg/l; Total-N, < 7 mg/l; NH4-N, < 1 mg/l. The change of process saved the community from a major investment in a separate treatment facility for the septic sludge. The stabilisation degree of the waste activated sludge is sufficient to by pass the anaerobic digestion for the time being. In the beginning, the SBR-process stability played an unwanted role during start up as it maintained a secondary phosphorus release for a considerable time. Later the process stability became an asset as the varying loads from the septic sludge addition were handled with very good results. PMID- 11381897 TI - Denitrifying phosphorus removal by anaerobic/anoxic sequencing batch reactor. AB - Denitrifying phosphorus removal was verified in a laboratory anaerobic/anoxic Sequencing Batch Reactor (A/A SBR) for a period of 18 months. The results obtained demonstrated the ability of the anaerobic/anoxic strategy to enrich the growth of denitrifying phosphorus bacteria (DPB) capable of taking up phosphate under anoxic conditions by using nitrate as the electron acceptor. Phosphorus removal efficiency ranging from 40-100% could be attained in an A/A SBR system. Simultaneous anoxic phosphate uptake and biological denitrification under anaerobic/anoxic condition occurred in this system. Batch tests showed, however, that simultaneous presence of carbon and nitrate would be detrimental to denitrifying phosphorus removal. Results of bacteria studies suggested that three denitrifying isolates had aerobic phosphorus removing ability. PMID- 11381898 TI - Effects of fill modes on N2O emission from the SBR treating domestic wastewater. AB - Nitrous oxide (N2O) gas is emitted as an intermediate in the biological nitrogen removal process. A track study was performed to investigate the characteristics of N2O emission depending on the cyclic mode of a sequencing batch reactor (SBR). A major emission of N2O took place at the aerobic phase, while N2O emission at the anoxic phase was insignificant. Especially, the highest N2O emission rate was observed at the initial stage of aerobic phase under the limited dissolved oxygen (DO) condition. Under such a condition, nitrite (NO2-) was transiently accumulated along with significant N2O emission due to incomplete nitrification. In addition, N2O production at the aerobic phase was strongly related with incomplete denitrification by nitrifiers. N2O emission could be reduced by change in fill modes in the SBR. A significant conversion to N2O took place in the SBR with the anoxic fill mode, while only small amount of N2O was conversed in the SBR with the aerobic fill mode. Relatively high concentration of ammonia nitrogen (NH4+) accelerated N2O production at the aerobic phase in the SBR with the anoxic fill compared to the aerobic fill. For control of N2O emission in the SBR, the aerobic fill mode could be an effective method even if denitrification efficiency may be reduced at the anoxic phase. PMID- 11381900 TI - Phosphorus removal from SBR with controlled denitrification for weak sewage. AB - Most SBRs operating for weak sewage have a tendency of poor removal of phosphorus due to NO3N inhibition. To minimize this inhibition, NO3N storage was utilized in this study. With influent BOD 100 mg/L and VFA 30 mg/L, a total of the maximum removable nitrogen and phosphorus was about 25 mg/L with 8 hr cycle. With this storage, VFA utilized for phosphorus removal could be maximized. The effluent nitrogen and phosphorus levels were generally 11 and 1 mg/L, respectively. PMID- 11381899 TI - Role of storage phenomena on removal of different substrates during pre denitrification. AB - Removal mechanisms of different substrates during the pre-denitrification step of an anoxic/aerobic sequencing process are studied. Biomass was cultivated in an anoxic/aerobic SBR and fed with a mixture of low and high molecular weight compounds. Substrate removal mechanisms are studied by means of batch tests, performed under anoxic conditions. The dynamic response to a spike of four different substrates (acetate, glucose, glutamic acid and ethanol) is described by simultaneously considering substrate and electron acceptor removal, and PHB and carbohydrates storage. PHB storage is a relevant mechanism during the removal of acetate and ethanol, while glucose is removed mainly by carbohydrate storage. PMID- 11381901 TI - Enhanced biological phosphorus removal in a semi full-scale SBBR. AB - A 17 m3 Sequencing Batch Biofilm Reactor (SBBR) was operated for enhanced biological phosphorus removal and nitrification for a period of 384 days. Enhanced biological phosphorus removal (EBPR) activity was instantly induced after start-up of EBPR operation mode and low phosphate effluent values were reached from the first batch onward. Process stability with regard to nitrification and EBPR were very good although high nitrate loads from backwashing disturbed the P removal performance. Due to anoxic conditions in the beginning of the cycle, readily degradable COD was depleted by denitrification. Consequently, particulate matter was the main carbon source for phosphorus accumulating organisms. Anaerobic hydrolysis or fermentation was found to be the rate limiting process in the SBBR cycle. Simultaneous denitrification occurred in the first 30 minutes of aeration and--to a lesser extent--during the remaining aeration time, enhancing nitrogen removal and indirectly also phosphorus removal. PMID- 11381902 TI - Modeling effect of remaining nitrate on phosphorus removal in SBR. AB - Nitrate shock loading experiments were conducted in a bench scale SBR to investigate the effect of nitrate on phosphorus removal. After achieving satisfactory phosphorus removal under steady state operation, initial NO3-N concentration amounting to 10 and 20 mg/L was fed at the beginning of the cycle. It was observed that, 10 mg/L of NO3-N suppressed phosphorus release during the feed and mix phases. Organic consumption for denitrification lead to limited PHA storage by phosphorus removing bacteria, resulting in less PO4-P removal. For 20 mg/L, influent organic substrate was not sufficient even for complete denitrification, thus leading to the presence of higher NO3-N and PO4-P in effluent. To explain the dynamics of the nutrient removal system under the transient loading, a SBR model based on IAWQ ASM2 was implemented. After adjusting PHA contents, model simulations well predicted dynamic changes of nitrate and phosphate concentrations during a cycle. Based on the model simulations, competition of COD substrate among denitrification, fermentation and oxygen respiration were investigated by calculating their consumption rates during mixing phase. In addition, a nitrate disappearance model was proposed and implemented in conjunction with a settling model to predict remaining and effluent nitrate in a cycle of SBR. Furthermore, integrated model simulations highlighted the effect of remaining nitrate on phosphorus release considering different options of reactions in settling phase. PMID- 11381903 TI - An enhanced biological phosphorus removal (EBPR) control strategy for sequencing batch reactors (SBRs). AB - A control strategy was developed for enhanced biological phosphorus removal (EBPR) in a Sequencing Batch Reactor (SBR). Unlike past research that focused on maximizing polyhydroxyalkanoate (PHA) formation during the anaerobic period, this study investigated some of the factors that govern aerobic PHA dynamics and its efficient regulation during phosphate (P) uptake. Influent COD, influent P, and the time for aeration were critical factors that governed PHA use and P uptake during aerated react. Unnecessary PHA oxidation (i.e., in the absence of extracellular P) occurred if the time for aerated react exceeded the time required for P uptake. By adjusting the aeration time to that required for P uptake, residual PHA was sustained in the SBR and excess phosphate uptake reaction potential (PRP) was generated for use during transient influent excursions in P. Unlike space oriented systems, the time for react is simply adjusted in the SBR. Because residual PHA is easily maintained once achieved, high influent COD events can be harnessed to increase or sustain excess PRP for management of expected variations in influent P. PMID- 11381904 TI - Generation and properties of aerobic granular sludge. AB - A sequencing batch reactor (SBR) was used to investigate the generation of different granules cultured under aerobic and alternating anaerobic/aerobic conditions. The reactor was fed with synthetic wastewater. A substrate loading rate of 3.6 kg COD/(m3 day) was applied. Granules of heterotrophic microorganisms were formed. After the first experimental period of 8 weeks the average granule diameter was 3.2 mm. In the second period, alternating anaerobic/aerobic conditions were applied to form granular sludge with an average diameter of 3.0 mm. An isopycnic centrifugation procedure was used to determine the characteristic density of the aerobic granular sludge. The average density of the granular sludge was 1.044 g/ml and 1.048 g/ml, respectively. In free-settling tests the final settling velocity of single aggregates was examined to estimate porosity. Settling velocities up to 2.0 cm/s could be measured. Calculations based on the experimental results showed an average granula porosity of 72% for the first run and 65% average porosity for the second run. This paper indicates the validity of general assumptions in free-settling tests. PMID- 11381905 TI - Biological denitrifying phosphorus removal in SBR: effect of added nitrate concentration and sludge retention time. AB - Optimizing anoxic biological phosphorus removal in the anaerobic-anoxic sequencing batch reactor (A2 SBR) was observed to depend on two parameters: the amount of added nitrate and the sludge retention time (SRT). The concentration of 120 mg N-NO3.l-1 in the anoxic medium and the SRT of 15 days were determined as optimal for a complete phosphorus removal in the A2 SBR. The reactor was supplied with synthetic wastewater containing 800 mg COD.l-1 acetic acid, 240 mg N-NH4.l-1 and 30 mg P-PO4.l-1. This study was completed by microscopic observations which revealed three morphological types of phosphate-accumulating bacteria (PAB). PMID- 11381906 TI - Sequencing batch reactor with submerged hollow fibre membranes for the biomass separation. AB - A method to reduce the cycle time of the SBR-process by utilising submerged membranes is described. The time necessary for the sedimentation of the activated sludge and the extraction of the clear-water phase takes up a major part of the entire cycle period within the SBR-process. By using membranes it is possible to start the clear-water extraction already during the reaction time. The time needed for the sedimentation of the activated sludge can be dispensed with altogether. The paper presents the initial results of a semi-technical test plant, with particular emphasis on the fact that the running of the SBR-plant did not lead to any problems with fouling or scaling. In cases of high solids contents, the oxygen input was below par; this, however, did not have any unfavourable impact on the nitrification. PMID- 11381907 TI - The anaerobic SBR process: basic principles for design and automation. AB - This study has determined the purification performance and the basic principles for the design of an anaerobic SBR (ASBR) to be used to treat wastewater generated in the food industries. Two ASBR's were set up and one fed with a slaughterhouse effluent at low concentration, the other with concentrated dairy wastewater. The maximum loading rate applied should not exceed 4.5 g of COD/L/day for the dilute effluent and 6 g of COD/L/day for the concentrated effluent. At higher loading rates, the reactors become difficult to operate, mainly because of sludge removal problems, and purification efficiency declines. A detailed study of the kinetics (TOC, VFA, rate of biogas production) throughout one treatment cycle led to the development of a simple control strategy based on the monitoring of the biogas production rate which was then applied to the reactor treating the dairy wastewater. After automation, the reactor worked free of problems at an average pollution load of 5.4 g of COD/L/day. PMID- 11381908 TI - SBR as a relevant technology to combine anaerobic digestion and denitrification in a single reactor. AB - Two laboratory-scale sequencing batch reactors were used to study biological treatment of carbon and nitrogen in a new combined anaerobic-aerobic system. Piggery wastewater was used as a model effluent. The anaerobic reactor, fed with raw wastewater and nitrite/nitrate recycling from the aerobic reactor, carried out denitrification and anaerobic digestion of organic carbon. In the aerobic reactor, organic carbon removal and nitrification of ammonia to mainly nitrite occurred. Three recycle-to-influent ratios from 1 to 3 were tested. The higher the recycle-to-influent ratio, the lower the concentrations of nitrogen oxides in the final effluent. Carbon removal efficiency was quite stable, whatever the ratio. However, the effect of this ratio was attenuated because of denitrification in the aerobic reactor, which increased the performances of the process. The use of sequencing batch reactors was essential to apply the configuration proposed, in order to combine denitrification and methanogenesis that require antagonistic conditions. PMID- 11381909 TI - Simulation and applications of a novel modified SBR system for biological nutrient removal. AB - Dynamic simulation and applications of a novel, continuous-fed, constant level modified sequencing batch reactor for biological nutrient removal are presented. The underlying mathematical model and practical applications of the simulation are discussed. Case studies are presented to illustrate the applications as well as the flexibility of the system in meeting different wastewater treatment requirements. Operation experience from full-scale wastewater treatment plant demonstrates the reliability, ease of operation and high efficiency of the system. Average BOD5, total nitrogen, total phosphorus and TSS removals of 97, 81, 88 and 94% are achieved respectively on an annual basis with little operator attention. Consistently high waste activated sludge concentrations are demonstrated, averaging approximately 20,000 mg/L. PMID- 11381910 TI - The effect of operating conditions on the performance of soil slurry-SBRs. AB - Biological treatment of a silty clay loam with aged diesel fuel contamination was conducted in 8 L Soil Slurry-Sequencing Batch Reactors (SS-SBRs). The purpose was to monitor slurry conditions and evaluate reactor performance for varying solids concentration (5%, 25%, 40%, 50%), mixing speed (300 rpm, 700 rpm, 1200 rpm), retention time (8 d, 10 d, 20 d), and volume replaced per cycle (10%, 50%, 90%). Diesel fuel was measured in slurry and in filtered aqueous samples. Oxygen uptake rate (OUR) was monitored. Aggregate size was measured with sieve analyses. Biosurfactant production was quantified with surface tension measurements. Increasing solids concentration and decreasing mixing speed resulted in increased aggregate size, which in turn increased effluent diesel fuel concentrations. Diesel fuel removal was unaffected by retention time and volume replaced per cycle. Biosurfactant production occurred with all operating strategies. Foam thickness was related to surfactant concentration and mixing speed. OUR, surfactant concentration, and foam thickness increased with increasing diesel fuel added per cycle. PMID- 11381911 TI - Period biofilter operation for enhanced performance during unsteady-state loading conditions. AB - In conventional biofilter operation, contaminated air is passed continuously through packed beds containing microbial consortia capable of contaminant biotransformation. This paper describes how biofilters can be designed and operated as controlled unsteady-state, periodic processes for the destruction of gas-phase contaminants. Such operation, previously limited to applications in wastewater treatment and soil remediation, increases an operator's ability to control the physiological state, "robustness," and spatial distribution of the microbial communities established within the biofilter and, thus, minimizes uncertainties that often accompany design and operation of biological systems. Results are presented from toluene degrading biofilters that used polyurethane foam packing medium. These studies demonstrate how controlled periodic operations can enhance contaminant removal during transient periods of elevated contaminant load. PMID- 11381912 TI - Intracellular dynamics of ribonucleic acid (RNA) and protein in microorganisms from periodically operated biofilters. AB - Conventional biofilters are designed and operated as continuous flow processes where the reactors receive a constant stream of contaminated air. Recent research has shown that periodically operated biofilters can remove a greater mass of contaminants during shock loads than equally sized continuously loaded biofilters. Preliminary experiments were conducted to investigate effects of periodic operation on physiological state of biofilter microorganisms. Relative concentrations of two macromolecular components of microbial cells, RNA and protein, were quantified in biosolids samples removed from biofilters operated under different periodic and continuous loading strategies. Preliminary studies presented herein suggest that the physiological state of the microbial population present in the periodically operated biofilter differs from that of those present in the biofilter operated continuously supplied air. PMID- 11381913 TI - Carbon and nitrogen removal from a wastewater of an industrial dairy laboratory with a coupled anaerobic filter-sequencing batch reactor system. AB - A set of two reactors, an Anaerobic Filter (AF) of 12 m3 and a Sequencing Batch Reactor (SBR) of 28 m3, coupled in series, were used to treat the wastewaters from an industrial milk analysis laboratory. The characteristics of these effluents are similar to those discharged by dairy factories (average values around 10 kg COD/m3 and 0.20 kg N/m3). These wastewaters were produced as the result of the final mixture of the analysed milk samples, with a very high organic load, and other low strength effluents, such as sewage and other minor liquid streams generated in the laboratory. Two microbial growth inhibitors, sodium azide and chloramphenicol, were systematically added to the milk before its analysis. Preliminary results have shown that these compounds did apparently not inhibit the methanogenic activity of the anaerobic sludge. Toxicity determination, using the Microtox method, resulted in EC50 values for the wastewaters of 20 g/L, whereas the final effluent from the SBR was non toxic. A maximum OLR of 8 kg COD/m3.d was treated in the AF, being the maximum OLR in the SBR around 1.5-2 kg COD/m3.d. During operation, the soluble COD of the final effluent from the SBR was usually below 200 mg/L, and total nitrogen (mainly nitrate) below 10 mg N/L. Assimilation of nitrogen for growth and nitrification denitrification were the main mechanisms of nitrogen removal from the wastewater. In the anaerobic system between 50-85% of the organic matter was converted into methane, being the remaining COD and most of the nitrogen removed in the suspended culture system. Overall COD removal in the treatment system was 98% and the nitrogen removal up to 99%. The combination of the AF and the SBR was advantageous resulting in a lower energy consumption and sludge generation in the treatment system. PMID- 11381914 TI - Modelling biological phosphorus removal from a cheese factory effluent by an SBR. AB - A mathematical model, named A3DX, based on ASM3(A3) for C and N removal, on the bio-P metabolic model of the Technological University of Delft (D), and on extra processes (X) for chemical and biological phosphorus removal, was developed and used to simulate the treatment of a fermented cheese factory effluent by a sequencing batch reactor (SBR). Experimental data obtained from a pilot-scale SBR were used to calibrate the model. The model calibration was performed by changing a minimal number (four) of default values for parameters, and by introducing a Monod function to account for magnesium limitation. This study suggests that the value of stoichiometric and kinetic model parameters determined with municipal effluents or enriched bio-P cultures can be reasonably used with some agro industrial effluents with minimal parameter adjustment for calibration. PMID- 11381915 TI - Characterization and treatment of the liquid effluents from the anaerobic digestion of biogenic solid waste. AB - The process wastewater generated by the fermentation of biogenic fractions of solid waste (biowaste) is characterized by very specific composition features, which sets it apart from the well known categories of wastewater. Up to now, there is a definite lack of practical experience and know-how in the treatment of those effluents. In view of the increasingly wide acceptance awarded to the fermentation process, it appears necessary, however, to include the treatment of the generated wastewater in the overall process and to grant it the same priority as the fermentation step. In this study, pilot-scale experiments were conducted over 200 days in a sequencing batch reactor (SBR), with the wastewater from a full-scale Bavarian biowaste digestion plant. Characterization of the influent implementing molecular size distribution studies showed that 97% of the components were either in particular form or < 1 kD. The main classes of compounds present in these fractions have been identified. The treatment objectives of the SBR plant were carbon and nitrogen removal. Soluble COD removal efficiencies were comprised between 40 and 60%, yielding a residual, refractory COD concentration of about 2000 mg/L in the effluent. Ammonium removal efficiencies averaged 96% and denitrification occurred with the addition of acetic acid or pretreated biowaste as a carbon source. Pretreated biowaste also showed synergetic effects as a co-substrate for COD removal. PMID- 11381916 TI - Use of image analysis and rheological studies for the control of settleability of filamentous bacteria: application in SBR reactor. AB - To monitor the ability of flocs to settle in Sequencing Batch Reactor sludge, two methods were tested during two operation cycles. Firstly, an automated image analysis procedure has been tested to quantify the floc size and the length and number of filaments. Secondly, rheological measurements (Bingham viscosity and shear stress) have been used to characterise the dispersion of the sludge which can reflect the cohesive strength of aggregates and so the influence of filamentous bacteria on rheological properties. These results were compared with settling parameters such as Sludge Volume Index or settling velocity. Correlations between the measured parameters with image analysis and parameters such as Sludge Volume Index have been obtained. If it is more difficult to analyze rheological results, it seems however that the thixotropy and the Bingham viscosity distinguish poor settlement owing to some filamentous bulking. The first results are promising, although they require confirmation in the long term. PMID- 11381917 TI - Creating anoxic and microaerobic conditions in sequencing batch reactors treating volatile BTX compounds. AB - An experimental strategy is introduced for studying the biodegradation of wastewaters containing volatile contaminants using an alternating anoxic/microaerobic sequencing batch reactor (SBR). Benzene, toluene, and the xylene isomers (BTX) served as model volatile contaminants for this study. The reactor was configured to overcome stripping the volatile BTX compounds into the atmosphere to provide opportunities for BTX biodegradation. Oxygen-free anoxic and microaerobic (< 0.2 mg/L dissolved oxygen) conditions were established using a novel laboratory reactor configuration. ORP was successfully used to monitor different electron acceptor conditions in the SBR. Toluene and m-xylene were amenable to anoxic (denitrifying) metabolism while benzene, o-, and p-xylene were biodegradable under microaerobic conditions. The results demonstrate that establishing microaerobic conditions in full-scale bioreactors may be an appropriate way to encourage the biodegradation of aerobically biodegradable volatile contaminants. Additionally, the laboratory reactor configuration introduced in this paper may be useful in subsequent studies involving microaerobic metabolism. PMID- 11381919 TI - Explosive biodegradation in soil slurry batch reactors amended with exogenous microorganisms. AB - The present study explores the feasibility of biotreatment of 2,4,6 trinitrotoluene (TNT) and hexahydro-1,3,5-trinitro- 1,3,5-triazine (RDX) contaminated soils in slurry batch reactors. Radiorespirometric assays showed that anaerobic sludge was able to mineralize 59% RDX to CO2 although significant mineralization of TNT was not observed in all cases. TNT and RDX at concentrations higher than 50 and 100 mg/L respectively were inhibitory to methanogenesis (used as a bioindicator), however, methanogenesis recovered after TNT was transformed into less toxic triaminotoluene. Bioslurry batch reactors containing 40% of contaminated soil (2000 mg RDX and 1000 mg TNT/kg dry soil) were operated under various conditions. Both TNT and RDX were persistent to soil indigenous microbes. Degradation of both TNT and RDX was enhanced by the municipal sludge amendment, although degradation of RDX was only achieved under anaerobic conditions. PMID- 11381918 TI - Strategies to enhance the biodegradation of toxic compounds using discontinuous processes. AB - This study presents two strategies used to enhance the biological degradation of phenolic wastewaters. In the first one the operation of a sequencing batch biofilter added with granular activated carbon (SBB-AC) was studied. The second strategy presents the results of the automation of a sequencing batch reactor in order to optimize the reaction phase. In this case, the dissolved oxygen was employed to monitor and control the reactor. The results of the SBB-AC system, based on the configuration of the reactor, type and size of activated carbon and size of the packing material, are discussed. The system biodegraded efficiently (total phenol removals as high as 97%) high concentrations (600 mg/l) of a mixture of phenol, 4-chlorophenol, 2,4-dichlorophenol and 2,4,6-trichlorophenol. Maximal eliminated loads of 4.33 kg COD/m3-d were achieved. For the second strategy, the applicability of an optimal control for a SBR using the dissolved oxygen as the measured variable was demonstrated. When the reactor was operated under the time-optimal control strategy, the degradation time of 4-chlorophenol was reduced. A very satisfactory operation of the reactor was observed, since the removal efficiencies were around 99%. PMID- 11381920 TI - Biodegradation of high strength phenolic wastewater using SBR. AB - This investigation demonstrates the capability of a bench-scale sequencing batch reactor (SBR) to biodegrade an inhibitory substrate at a high loading rate. A SBR loading rate of 3.12 kg phenol.m3 d-1 (2.1 g COD.g-1 MLVSS d-1) with a COD removal efficiency of 97% at a SRT of 4 days and a HRT of 10 hours was achieved; this rate was not reached before. The SBR was operated at 4 hours cycle, including 3 hours react phase. The synthetic wastewater of 1300 mg/L phenol was the sole carbon source. Oxygen uptake rates (OUR) were monitored in-situ at various stages of the SBR. The oxygen mass transfer coefficient, KLa, of 12.6 h-1 was derived from respirometry. Use of respirometry in SBR aided the tracking of the soluble substrate through OUR. PMID- 11381921 TI - Biological ammonia removal from anaerobically pre-treated landfill leachate in sequencing batch reactors (SBR). AB - The aim of the study was to investigate both the use of the SBR technology in biological ammonia removal from landfill leachate, and the suitability of raw landfill leachate as external carbon source in denitrification step. The SBR was fed with diluted leachate for the first 42 days and then the effluent of UASBR was used as the feed. The SBR was operated intermittently with a cycle time of 24 hours. The effluent NH4(+)-N values of less than 5 mg NH4(+)-N L-1 was consistently observed for the initial NH4(+)-N levels of as high as 1000 mg NH4(+)-N L-1. The nitrification rates for the first, second and third stages were found as 5.7, 46.8 and 102.8 mg NH4(+)-N L-1 h-1, respectively. The difference of the nitrification rates in the 2nd and 3rd stages originated from increasing adaption of the sludge as well as increasing biomass concentration (10.5 mg NH4(+)-N g-1 VSS h-1). No significant accumulation of NO2(-)-N has been observed during the study and NO2(-)-N/NOx(-)-N ratios measured in the 1st aerobic phase and the SBR effluent were less than 7%. The denitrification rates for the second (raw leachate as carbon source) and the third (Ca(CH3COO)2 as carbon source) stages were determined as 45.7 mg NOx(-)-N L-1 h-1 (or 9.85 mg NOx(-)-N g-1 VSS h 1) and 125.7 mg NOx(-)-N L-1 h-1 (or 12.88 mg NOx(-)-N g-1 VSS h-1), respectively. PMID- 11381922 TI - Exceptionally high-rate nitrification in sequencing batch reactors treating high ammonia landfill leachate. AB - The nitrogen removal capacity of a suspended culture system treating mature landfill leachate was investigated. Leachate containing high ammonium levels of 300-900 mg N/L was nitrified in a bench scale sequencing batch reactor. Leachate from four different landfills was treated over a two year period for the removal of nitrogen. In this time, a highly specific nitrifying culture was attained that delivered exceptionally high rates of ammonia removal. No sludge was wasted from the system to increase the throughput and up to 13 g/L of MLSS was obtained. Settleability of the purely nitrifying biomass was excellent with SVI less than 40 mL/g, even at the high sludge concentrations. Nitrification rates up to 246 mg N/(L h) (5.91 g N/(L d)) and specific nitrification rates of 36 mg N/(gVSS h) (880 mg N/(gVSS d)) were obtained. The loading to the system at this time allowed complete nitrification of the leachate with a hydraulic retention time of only 5 hours. Following these successful treatability studies, a full-scale plant was designed and built at one of the landfills investigated. PMID- 11381923 TI - SBR technology in Germany--an overview. AB - The SBR technology is applied in about 1.3% of the wastewater treatment plants (WWTP) in Germany. This caused the German Association for the Water Environment (ATV) to prepare the guideline ATV-M 210 to represent the state of the art for this type of WWTPs in Germany. The basic design parameters were derived from the standard ATV-A 131 for activated sludge plants to prepare a basis for the comparison of alternatives. In Bavaria numerous small WWTPs utilize SBR technology. Operational experiences show that these plants require specifically trained personnel. The plants produce effluents comparable to those of continuous flow plants in every respect. Hence the choice of system can be based on economic criteria. PMID- 11381924 TI - Cyclic activated sludge technology--recent operating experience with a 90,000 p.e. plant in Germany. AB - Cyclic activated sludge technology was selected for the Potsdam Wastewater Treatment Plant (90,000 p.e.). The cyclic activated sludge facility comprises four modules integrated into two circular basins. Construction was commenced in February 1998 with seeding of the plant for start up taking place in October 1998. Process performance has been met since Spring 1999 at 80-90% of design load. In order to optimize start-up procedures, respiration rates were used as a guidance for process stabilization and online process optimization during normal operation. Operation for co-current nitrification denitrification provided an ammonia removal of 1.1 mg NH4-N/g MLSS.h (15 degrees C) and a corresponding nitrate respiration rate of 0.85 mg NO3-N/g MLSS.h under aerated conditions. Enhanced biological phosphorus removal generated an effluent mean total phosphorus concentration of 0.38 mg/L without precipitant addition. PMID- 11381925 TI - Sequencing batch reactor technology: the key to a BP refinery (Bulwer Island) upgraded environmental protection system--a low cost lagoon based retro-fit. AB - BP Refinery (Bulwer Island) Ltd (BP) located on the eastern Australian coast is currently undergoing a major expansion as a part of the Queensland Clean Fuels Project. The associated wastewater treatment plant upgrade will provide a better quality of treated effluent than is currently possible with the existing infrastructure, and which will be of a sufficiently high standard to meet not only the requirements of imposed environmental legislation but also BP's environmental objectives. A number of challenges were faced when considering the upgrade, particularly; cost constraints and limited plot space, highly variable wastewater, toxicity issues, and limited hydraulic head. Sequencing Batch Reactor (SBR) Technology was chosen for the lagoon upgrade based on the following; SBR technology allowed a retro-fit of the existing earthen lagoon without the need for any additional substantial concrete structures, a dual lagoon system allowed partial treatment of wastewaters during construction, SBRs give substantial process flexibility, SBRs have the ability to easily modify process parameters without any physical modifications, and significant cost benefits. This paper presents the background to this application, an outline of laboratory studies carried out on the wastewater and details the full scale design issues and methods for providing a cost effective, efficient treatment system using the existing lagoon system. PMID- 11381926 TI - From the lab to full-scale SBR operation: treating high strength and variable industrial wastewaters. AB - This paper describes the path taken from client objectives through laboratory studies and detailed design to full-scale SBR operation and current research. Conventional municipal design principles have often been used to develop treatment processes for industrial wastewaters. The use of scientific trials to test design criteria offers the client a "tailor made" design fir for their particular wastewater character. In this project, a waste management company wished to upgrade their physical-chemical treatment plant to incorporate a biological reactor for treating a range of industrial wastewaters. Laboratory scale trials were undertaken to determine appropriate design criteria for a full scale biological process. These laboratory studies indicated that conventional design criteria were not appropriate and that a SBR configuration was optimal compared with an IDAR configuration. It was also found that a novel fungal:bacterial mixed liquor consortium developed, resulting in good effluent quality and settling properties. The treatment plant was able to be constructed and operational within a tight timeframe and budget, allowing the client to take advantage of a commercial opportunity. The plant has been operating since 1997 and meets its discharge conditions. By combining scientific studies with engineering principles, the end-user obtained a complete treatment plant to meet their specific needs. A further benefit of the laboratory trials is current research into the development of a fungal:bacterial SBR to treat industrial wastewaters. This offers ongoing knowledge to the operational full-scale SBR. PMID- 11381927 TI - Using the flexibility index to compare batch and continuous activated sludge processes. AB - This paper considers the question of which is better: the batch or the continuous activated sludge processes? It is an important question because dissension still exists in the wastewater industry as to the relative merits of each of the processes. A review of perceived differences in the processes from the point of view of two related disciplines, process engineering and biotechnology, is presented together with the results of previous comparative studies. These reviews highlight possible areas where more understanding is required. This is provided in the paper by application of the flexibility index to two case studies. The flexibility index is a useful process design tool that measures the ability of the process to cope with long term changes in operation. PMID- 11381928 TI - Full-scale demonstration of biological nutrient removal in a single tank SBR process. AB - Complete biological nutrient removal (BNR) in a single tank, sequencing batch reactor (SBR) process, is demonstrated here at full-scale on a typical domestic wastewater. The unique feature of the UniFed process is the introduction of the influent into the settled sludge blanket during the settling and decant periods of the SBR operation. This achieves suitable conditions for denitrification and anaerobic phosphate release which is critical to successful biological phosphorus removal. It also achieves a "selector" effect, which helps in generating a compact, well settling biomass in the reactor. The results of this demonstration show that it is possible to achieve well over 90% removal of COD, nitrogen and phosphorus in such a process. Effluent quality achieved over a six-month operating period directly after commissioning was: 29 mg/l COD, 0.5 mg/l NH4-N 1.5 mg/l NOx-N and 1.5 mg/l PO4-P (50%-iles of daily samples). During an 8-day, intensive sampling period, the effluent BOD5 was < 2 mg/l in all samples and the total phosphorus averaged 0.17 mg/l in the effluent. Detailed sampling and analysis during one cycle and at various depths clearly showed the deliberate stratification achieved in the tank during the settling and decant period, allowing biochemical reactions to occur during this normally "non-productive" period. The simplicity and flexibility of the UniFed system allows it to be used in numerous applications, particularly for industrial situations where a high degree of uncertainty of the wastewater composition during the design stage or where changing requirements based on changes on the production side are present. The single tank operation without any recycle also reduces the capital costs for a full BNR system compared to the comparatively complex continuous flow processes. PMID- 11381929 TI - Biological nutrient removal in a full-scale SBR treating piggery wastewater: results and modelling. AB - Research activities carried out at ENEA during the last few years allowed development of a Sequencing Batch Reactor (SBR) that is able to remove biologically organic waste, nitrogen and phosphorus and that showed to be particularly suited to obtain low effluent nutrient concentrations even starting from concentrated wastes. This plant, in more than one year of operation, is quite steadily obtaining more than 98% removal of nitrogen, phosphorus and COD. On the basis of the experimental results, a simulation model has been built and calibrated. The model showed the potential to be used for forecasting the behaviour of the process, being able to reproduce a process imbalance that followed the tentative reduction of aeration time. PMID- 11381930 TI - The SBR process: an efficient and economic solution for the treatment of wastewater at small cheesemaking dairies in the Jura mountains. AB - The small cheesemaking plants in the Jura Mountains (N.E. France) need self contained solutions for treating their wastewater. Any solution must suit the characteristics of the effluent produced by these small production units and be simple, robust and financially feasible in regard to both capital and running costs. Given this context, a technique based on an SBR was recommended and the operations of the first treatment plant established on an industrial scale, monitored for the first year, with particular attention paid to operating constraints. The results obtained show that the SBR is, from a technical point of view, perfectly adapted to treating cheese production wastewater, with purification levels at 97.7% for total COD and 99.8% for BOD5. In the course of this study, it has been shown that the SBR process, thanks to its simple design and operation, can easily be run by the cheesemaker who will need to devote a minimum of time to it. Furthermore, with treatment costs of around 2 centimes (0.33 US cent) per litre of milk delivered, the SBR process meets perfectly both the technical and financial conditions for treatment as laid down by the Federation of Cooperatives of the Doubs and Jura departements. PMID- 11381931 TI - Comparative investigations on COD-removal in sequencing batch reactors and continuous flow plants. AB - Investigations on enhanced COD removal from municipal wastewater were performed over a period of 2.5 years, comparing three different types of reactor. The main idea was to determine the influence of the mixing characteristics of the reactor on the treatment processes and the effluent quality. Therefore three pilot plants (a completely mixed reactor, a cascade of three reactors and a SBR) were operated under equal conditions (wastewater, hydraulic load, temperature, sludge age) in parallel to each other. Investigations were carried out at different sludge ages. It could be shown that within one sludge age the CODf removal efficiency increased, when mixing characteristics came closer to the plug flow and it also increased with higher temperatures. A significant correlation was observed between the COD removal efficiency and the sludge load. The higher the sludge load was the greater the CODf concentration in the effluent. Especially the SBR reactor showed an excellent performance under the given operating conditions. Dynamic simulation calculations were carried out, to investigate whether the influence of the type of reactor on the COD-elimination could be described theoretically by combining growth kinetics and the mixing characteristics of the individual reactors. The results showed that performance was better when mixing characteristics came closer to plug flow. PMID- 11381932 TI - The mechanism and design of sequencing batch reactor systems for nutrient removal -the state of the art. AB - The Sequencing Batch Reactor (SBR) process for carbon and nutrient removal is subject to extensive research, and it is finding a wider application in full scale installations. Despite the growing popularity, however, a widely accepted approach to process analysis and modeling, a unified design basis, and even a common terminology are still lacking; this situation is now regarded as the major obstacle hindering broader practical application of the SBR. In this paper a rational dimensioning approach is proposed for nutrient removal SBRs based on scientific information on process stoichiometry and modelling, also emphasizing practical constraints in design and operation. PMID- 11381933 TI - Use of computer simulation for cycle length adjustment in sequencing batch reactor. AB - The city of Nowy Targ located in Poland's highland operates the WWTP with classic SBR technology. In winter some decrease in process efficiency was observed. The research described in the paper was aimed at using computer simulation to adjust SBR cycle length and structure in order to minimize the effects of low wastewater temperature on biological CNP removal. The simulations performed at 6 degrees C wastewater temperature showed that switching from 6 to 8 hours cycle and extension of the aeration phase length from 2.75 to 5 hours will increase the effectiveness of biological nitrification and biological phosphorus removal. The change in practice will not affect COD value in the effluent. Based on the simulation results a new SBR operational strategy was proposed for Nowy Targ WWTP. PMID- 11381934 TI - Implementation, study and calibration of a modified ASM2d for the simulation of SBR processes. AB - An enhanced process model for SBRs has been developed. Though the basic mechanism largely draws on the Activated Sludge Model n. 2d, its new features are the splitting of the nitrification stage in a two-step process, according to the well known Nitrosomonas-Nitrobacter oxidation sequence, and an improved XPAO dynamics, involved in the anaerobic/aerobic phosphorus removal process. The model was implemented through the DLL technique allowing complied C++ modules to be linked to an ordinary Simulink block diagram. The static sensitivity study revealed that if the parameter vector is partitioned into subsets of biologically related parameters and calibrated separately, the calibration procedure does not present particularly difficult aspects. Trajectory sensitivity showed also to which extent data collection could be optimised in order to improve calibration accuracy. The study of the shape of the error functional generated by parameters couples allows a much more effective calibration strategy. PMID- 11381935 TI - Performance of triple ditch: effects of duration and periodic operation programs. AB - The periodic variation of operation states in triple ditch (BIO-DENITRO process) was investigated in a pilot-scale and a full-scale triple ditch treating raw wastewater containing 70% petrochemical wastewater and 30% domestic wastewater. Mathematical models describing the periodic variation of activated sludge concentrations in each ditch were proposed based on theoretical analysis and were verified in the pilot-scale and full-scale triple ditches respectively. The existence of optimal cycle time and time arrangement of periodic operation programs were demonstrated and discussed according to the mathematical models and the experimental results of the pilot-scale triple ditch operated in four different stages. Four rulers determining the duration and operation programs are suggested. PMID- 11381936 TI - Feedforward aeration control of a Biocos wastewater treatment plant. AB - The Biocos strategy as a cyclical time controlled activated sludge system shows a great variability in operation and control. One topic such a type of treatment plant has to deal with is the optimum relation between aerobic and anoxic conditions. The aeration control has to adapt the length of the nitrification phases to the current constraints in order to save operational costs and maximise nitrogen elimination. Since wastewater treatment plants up to a certain size are usually not equipped with on-line nitrogen probes, influent flow and temperature can be taken as control parameters for the aeration system. The defined relation between influent flow and ammonia load is based on measurements and the relation between ammonia load and required aeration time is model based. PMID- 11381937 TI - Nitrification in sequencing biofilm batch reactors: lessons from molecular approaches. AB - The nitrifying microbial diversity and population structure of a sequencing biofilm batch reactor receiving sewage with high ammonia and salt concentrations (SBBR 1) was analyzed by the full-cycle rRNA approach. The diversity of ammonia oxidizers in this reactor was additionally investigated using comparative sequence analysis of a gene fragment of the ammonia monooxygenase (amoA), which represents a key enzyme of all ammonia-oxidizers. Despite the "extreme" conditions in the reactor, a surprisingly high diversity of ammonia- and nitrite oxidizers was observed to occur within the biofilm. In addition, molecular evidence for the existence of novel ammonia-oxidizers is presented. Quantification of ammonia- and nitrite-oxidizers in the biofilm by Fluorescent In situ Hybridization (FISH) and digital image analysis revealed that ammonia oxidizers occurred in higher cell numbers and occupied a considerably larger share of the total biovolume than nitrite-oxidizing bacteria. In addition, ammonia oxidation rates per cell were calculated for different WWTPs following the quantification of ammonia-oxidizers by competitive PCR of an amoA gene fragment. The morphology of nitrite-oxidizing, unculturable Nitrospira-like bacteria was studied using FISH, confocal laser scanning microscopy (CLSM) and three-dimensional visualization. Thereby, a complex network of microchannels and cavities was detected within microcolonies of Nitrospira-like bacteria. Microautoradiography combined with FISH was applied to investigate the ability of these organisms to use CO2 as carbon source and to take up other organic substrates under varying conditions. Implications of the obtained results for fundamental understanding of the microbial ecology of nitrifiers as well as for future improvement of nutrient removal in wastewater treatment plants (WWTPs) are discussed. PMID- 11381938 TI - On-line control of a SBR system for nitrogen removal from industrial wastewater. AB - In this study, laboratory-scale experiments were conducted applying the SBR activated sludge process to a wastewater from a wood factory operating on plywood production. Due to the fact that the wastewater is characterised with a high concentration of ammonia, the aim was to evaluate the nitrogen removal process with SBR system operating with a on-line control of ORP, pH and DO. The complete ammonia removal corresponded exactly to the "Ammonia Valley" in the pH versus time graphic. The ammonia removal efficiency in SBR was 99% at 20 degrees C. The denitrification reaction was completed when in the ORP versus time graphic appeared the "Nitrate Knee". From the experimental results the optimisation of the SBR cycle allowed the doubling of the applied load with respect to a not optimised cycle (performed in the same volume reactor), without a decrease in the removal efficiency. Moreover, the possibility of stopping the aeration just after the appearance of the Ammonia Valley allows an energy saving. The easy operation and the low cost make the SBR system an interesting option for the biological pre treatment of plywood industry to be discharged into surface water. PMID- 11381939 TI - Radioecology--how did we get here and what do we do now? PMID- 11381940 TI - Variability in the edible fraction content of 60Co, 99Tc, 110mAg, 137Cs and 241Am between individual crabs and lobsters from Sellafield (north eastern Irish Sea). AB - We investigated the variability of anthropogenic radionuclide content of the edible fractions of individual edible crabs (Cancer pagurus L.) and European lobsters (Homarus gammarus L.) caught commercially in the Sellafield offshore area. Sixteen female and 18 male crabs and 20 female and 17 male lobsters were selected from commercial catches made between 25 May and 5 June 1997. Each gender group was selected to be within the known weight range for commercially caught crustacea from the area. Four artificial radionuclides (60Co, 110mAg, 137Cs or 241Am) were detected by gamma-spectrometry. The edible fraction content of these radionuclides between males and females for either species were not statistically significantly different. 99Tc was analysed by chemical separation and beta counting. 99Tc concentrations in female crabs tended to be higher (172 +/- 205 (16) Bq kg-1(wet); mean +/- standard deviation (n samples)) than those in males (85 +/- 58 (18) Bq kg-1 (wet)), although this was not a statistically significant difference. For both male and female crabs, 99Tc concentrations tended to decrease with increasing whole live weights. For 99Tc in lobsters the picture is less clear. Female lobsters contained more activity (14800 +/- 7400 (20) Bq kg-1 (wet)) than males (7100 +/- 3900 (17) Bq kg-1 (wet)). The results were used to discuss the implications for sampling and monitoring. PMID- 11381941 TI - A blind test of the MOIRA lake model for radiocesium for Lake Uruskul, Russia, contaminated by fallout from the Kyshtym accident in 1957. AB - This paper presents results of a model-test carried out within the framework of the COMETES project (EU). The tested model is a new lake model for radiocesium to be used within the MOIRA decision support system (DSS; MOIRA and COMETES are acronyms for EU-projects). This model has previously been validated against independent data from many lakes covering a wide domain of lake characteristics and been demonstrated to yield excellent predictive power (see Hakanson, Modelling Radiocesium in Lakes and Coastal Areas. Kluwer, Dordrecht, 2000, 215 pp). However, the model has not been tested before for cases other than those related to the Chernobyl fallout in 1986, nor for lakes from this part of the world (Southern Urals) and nor for situations with such heavy fallout as this. The aims of this work were: (1) to carry out a blind test of the model for the case of continental Lake Uruskul, heavily contaminated with 90Sr and 137Cs as a result of the Kyshtym radiation accident (29 September 1957) in the Southern Urals, Russia, and (2) if these tests gave satisfactory results to reconstruct the radiocesium dynamics for fish, water and sediments in the lake. Can the model provide meaningful predictions in a situation such as this? The answer is yes, although there are reservations due to the scarcity of reliable empirical data. From the modelling calculations, it may be noted that the maximum levels of 137Cs in fish (here 400 g ww goldfish), water and sediments were about 100,000 Bq/kg ww, 600 Bq/l and 30,000 Bq/kg dw, respectively. The values in fish are comparable to or higher than the levels in fish in the cooling pond of the Chernobyl NPP. The model also predicts an interesting seasonal pattern in 137Cs levels in sediments. There is also a characteristic "three phase" development for the 137Cs levels in fish: first an initial stage when the 137Cs concentrations in fish approach a maximum value, then a phase with relatively short ecological half lives followed by a final phase with long ecological half-lives more or less corresponding to the physical decay of radiocesium. PMID- 11381942 TI - A mechanistic sub-model predicting the influence of potassium on radiocesium uptake in aquatic biota. AB - It is often argued that the quality of science is related to the possibilities of making accurate predictions. It has also long been argued that due to the complex nature of ecosystems, it will never be possible to predict important target variables, especially with more comprehensive dynamic models. New results in radioecology have, however, demonstrated that those arguments are no longer valid. The key to the predictive success lies in the structuring of the model. The accident at Chernobyl has, in fact, provided science with an intriguing opportunity to study how the pulse of 137Cs is transported through ecosystem pathways, thus revealing the basic structure of these ecosystems, i.e. which are the key and the less-important pathways. It is paradoxical to conclude that the Chernobyl accident is, perhaps, the most important factor behind the revolution in predictive ecosystem modelling which lies behind the decrease in the uncertainty factor from 10 to 0.25-0.5. The sub-model for the potassium moderator presented in this paper is an example of a mechanistically based sub-model used within the framework of a more comprehensive lake model for 137Cs. The K moderator presented is derived from the existing knowledge on ion transport in biological membranes and takes into account ion equilibria modelled by the Nernst equation and the uptake kinetics quantified by the Michaelis-Menten model. It provides the type of structure to this overall lake model that helps to explain the excellent predictive power of this model. PMID- 11381943 TI - Sorption and transport of radionuclides by tumbleweeds from two plastic-lined radioactive waste ponds. AB - Previous research has examined the uptake of radionuclides by tumbleweeds growing in contaminated soils, but none has heretofore examined the sorption of radionuclides to tumbleweeds blowing into radioactively contaminated water. Three tumbleweed species; Russian thistle (Salsola kali), Jim Hill mustard (Sisymbrium altissimum) and summer cypress (Kochia scoparia) blow in, and out of, two plastic lined radioactive wastewater ponds, constructed in 1993 on the US Department of Energy's Idaho National Engineering and Environmental Laboratory in southeast Idaho. This research quantified radionuclide sorption to tumbleweeds, tumbleweed movement from the ponds, and determined radionuclide transport from the ponds. Average plant/water concentration factors associated with tumbleweeds taken from the ponds ranged from 5 for 152Eu to over 9000 for 54Mn. Based on changes in tumbleweed numbers and average concentrations associated with them, 66.2 MBq were estimated to have been transported from the ponds via tumbleweeds between 18 October 1994 and 8 November 1996. This amounts to about 0.01% of the non-tritium and 0.0002% of the tritium activity released to the ponds through 8 November 1996. A power function best described the radionuclide buildup curve for tumbleweeds submerged in the ponds. Visually marked tumbleweeds traveled from the ponds in the predominant wind direction a maximum of 737 m. Management practices which may reduce the number of tumbleweeds blowing both into and out of contaminated ponds are discussed. PMID- 11381944 TI - Improvements in modeling sagebrush concentrations of radioiodine released from the Hanford site. AB - The Hanford Environmental Dose Reconstruction Project estimated the radiation dose to individuals from historical emissions of radioactive materials from the Hanford Site in Washington State. Project validation studies using predicted activity concentrations of 131I on sagebrush (Artemisia tridentata) showed a systematic underestimation against historical data during cold weather months, indicating a need for sagebrush model improvement. A deposition model for semi volatile organic materials presented by Komp and McLachlen (Environ. Sci. Tech. 31 (1997) 886-890) is adapted here for gaseous iodine onto sagebrush. The deposition model includes a temperature-dependent term based on an integrated van't Hoff equation. Calibration data for the model are obtained from a release of 131I in 1963. Modeling results for releases in 1946 show a good match between historical data and predicted results using the new model. The new model shows improvement over interception-fraction type models, but requires plant-type specific calibration data. PMID- 11381945 TI - Distribution of 137Cs in soil along Ta-han River Valley in Tau-Yuan County in Taiwan. AB - Environmental 137Cs contamination was suspected from accidents at spent fuel storage pits of a research reactor site in the Ta-han River valley in Taiwan. In order to further characterize this contamination, soil samples were collected and measured by a gamma-spectroscopy system in 1999. It was found that 137Cs contamination is distributed up to 4 km from the reactor in an area covered mostly by rice and plant fields. 137Cs concentration in the topsoil ranged up to about 1000 Bq kg-1, as compared with soil beyond the contaminated area, which does not exceed 15 Bq kg-1. Spatial distribution of 137Cs was characterized by strong non-uniformity, which complicated our understanding of the distribution pathway of the radionuclides. The highest concentrations of 137Cs, up to more than 1000 Bq kg-1, were found within a few rice fields. The relative location of these rice fields and the water supplies from local streams suggested that the 137Cs was distributed along water pathways in the valley. PMID- 11381946 TI - Radon emanation from NORM-contaminated pipe scale and soil at petroleum industry sites. AB - Radon-222 emanation fractions were determined for barite scale deposits associated with petroleum production tubing and soil contaminated with naturally occurring radioactive material (NORM). Samples were analyzed for 226Ra concentration, the results of which were used to calculate the 222Rn emanation fraction for the sample. An important parameter determining the overall Rn activity flux from a solid medium, 222Rn emanation fraction represents the fraction of 222Rn produced that enters the interconnected pore space within a medium contaminated with 226Ra before the 222Rn undergoes radioactive decay. The primary objective of the study was to determine whether 222Rn emanation fractions from pipe scale and soil from petroleum production sites are similar to those of uranium mill tailings. Pipe scale samples were collected at four sites representing a wide geographical area, and consisted primarily of barite scale where Ra atoms have replaced a fraction of the Ba within the crystal lattice of the scale. Soil samples were collected at five sites, from areas exhibiting elevated surface gamma exposure rates indicating the presence of NORM. For comparison, 226Ra concentrations and 222Rn emanation fraction were also determined for uranium mill tailings samples provided from a site in Utah. Although 2226Ra concentrations from pipe scale samples were similar to those found in uranium mill tailings, 222Rn emanation fractions from scale were generally lower. Emanation fractions from each data set were statistically different from those of mill tailings (p < or = 0.01). The differences are probably due to physical differences between the two media and to the method by which the Ra is deposited in the material. Radon emanation from soils was extremely variable owing not only to differences in physical and chemical soil properties, but also to the means by which NORM has entered the soil. Although additional emanation measurements from other sites are needed, the data collected at these sites indicate that regulations intended to protect human health from 222Rn inhalation should consider the type and properties of the medium in which the NORM is contained, rather than relying strictly on concentrations of the parent 226Ra. PMID- 11381947 TI - Estimation of radon concentrations in coal mines using a hybrid technique calibration curve. AB - The results of epidemiological studies in various countries show that radon and its progeny cause carcinogenic effects on mine workers. Therefore, it becomes of paramount importance to monitor radon concentrations and consequently determine the radon dose rates in coal mines for the protection of coal miners. A new calibration curve was obtained for radon concentration estimation using hybrid techniques. A calibration curve was generated using 226Ra activity concentration measured by a HPGe detector-based gamma-ray spectrometer versus alpha-track density rate due to radon and its progeny on CR-39 track detector. Using the slope of the experimentally determined curve in the units of Becqueral per kilogram (Bq kg-1) per unit alpha-track-density per hour (cm-2 h-1), radon concentrations (Bq m-3) were estimated using coal samples from various coal mines in two provinces of Pakistan, Punjab and Balochistan. Consequently, radon dose rates were computed in the simulated environment of the coal mines. Results of these computations may be considered with a caveat that the method developed in this paper provides only a screening method to indicate the radon dose in coal mines. It has been shown that the actual measurements of radon concentrations in the coal mines are in agreement with the estimated radon concentrations using the hybrid-technique calibration curve. PMID- 11381948 TI - An integrated assessment of long-term changes in the hydrology of three lowland rivers in eastern England. AB - The flow records of the Rivers Bure, Nar and Wensum in eastern England have been examined with the aim of identifying long-term changes in flow behaviour relating to variations in rainfall amount, land use, land drainage intensity and water resources use. In the study area, and since 1931, there is no evidence of long term change in rainfall amount or distribution, on either an annual or seasonal basis. Despite changes in water resources use and catchment characteristics since the beginning of the century, such as the ending of water milling and increased land drainage and arable farming, rainfall-runoff modelling over the period 1964 1992 showed that the relationship between rainfall and runoff has remained essentially unchanged in the three study rivers. A catchment resource model used to 'naturalise' the historic flow records for the period 1971-1992 to account for the net effect of water supply abstractions and discharges revealed that mean river flows have been altered by surface water and groundwater abstractions, although the average losses to mean weekly flows due to net abstractions for all water uses was no greater than 3%. Greater losses occurred during drought periods as a result of increased consumptive use of water for spray irrigation and amounted to a maximum loss of 24% in the Nar catchment. In lowland areas such as eastern England that are prone to summer dry weather and periodic drought conditions, an integrated approach to river basin management, as advocated by the EU Framework Directive, is recommended for future management of surface and groundwater resources for public water supplies, river regulation purposes and industrial and agricultural demands. PMID- 11381949 TI - A quantitative evaluation of monitoring networks for region-specific nitrate reduction policies. AB - Modern environmental policy implementation in many developed countries is increasingly regionally oriented. Regional governments have undertaken measures designed for the specific needs of the region but, so far, the resulting change in environmental quality has hardly been monitored. This study develops a method for the quantitative evaluation of region-specific environmental policy. Two forms of monitoring are distinguished: monitoring of performance of policy measures and monitoring of environmental quality. This study investigates two regions in the Netherlands where region-specific policy has been implemented. Performance monitoring focused on reduction of nitrate leaching to groundwater. Leaching reductions were calculated with simple models or rules of thumb. Quality monitoring focused on nitrate concentrations under grassland and other land use. An analysis of variance showed significant differences between different soil units. Stratified kriging was applied for spatial interpolation, showing within unit variability. The current monitoring network could be improved by using a more regular type of grid. PMID- 11381950 TI - Local people's perceptions of planning and management issues in Prespes Lakes National Park, Greece. AB - Local people's perceptions of planning and management issues were investigated in Prespes Lakes National Park in north-western Greece, 24 years after designation. Ensued conflicts due to lack of local community participation in the designation procedure and in the decision-making process thereafter necessitated this research. Knowledge of the park and its aims, source of information about aims, necessity for works and facilities, attitudes toward certain policies, and effectiveness of administration and management scheme, were studied by means of a questionnaire survey. Respondents were contacted by systematic sampling, which resulted in 201 cases for analysis. Poor knowledge of aims associated with education of people was revealed and the managing authority (the Forest Service) as source of information was mentioned in only one case. Forest recreation facilities and improvement of accessibility were considered of high priority, as means of possible tourism development of the area. A policy of non-intensive agriculture with compensation for loss of income, if the wetlands of the park were in danger, seems acceptable, younger ages accepting it more easily. The need for a new administration and management scheme with the participation of local communities in the decision-making process was revealed, supported mainly by the younger age groups. Finally, the results indicated that the information derived from such research could help managers of protected areas to resolve arising conflicts. PMID- 11381951 TI - Recovery of big sagebrush communities after burning in south-western Montana. AB - Prescribed burning of big sagebrush (Artemisia tridentata Nutt.) communities is conducted with the intention of increasing either the productivity of the understory plants or the big sagebrush. It was our objective to compare the recovery of big sagebrush communities from prescribed fire at as many sites as we could locate in south-western Montana with environmentally paired unburned portions. We located and sampled 13 sites that had been burned over a span of two to 32 growing seasons earlier. Big sagebrush canopy cover, density, and production of winter forage were significantly greater (P < or = 0.05) in the unburned portions in 34 of 38 comparisons. Canopy coverage of Idaho fescue (Festuca idahoensis Elmer), the dominant herbaceous species, was greater in the burned portion at only one site while it was less (P < or = 0.05) at four sites. Total perennial grass canopy coverage was not different (P < or = 0.05) between treatments over the 13 sites. Managers considering prescribed burning of big sagebrush communities should be aware that herbaceous plant responses may be minimal while shrub values will likely be lost for many years. The loss of the dominant shrubs in any ecosystem will affect many other organisms and severely impact species that have an obligate habit with the shrubs. PMID- 11381952 TI - Transaction costs and sequential bargaining in transferable discharge permit markets. AB - Market-type mechanisms have been introduced and are being explored for various environmental programs. Several existing programs, however, have not attained the cost savings that were initially projected. Modeling that acknowledges the role of transactions costs and the discrete, bilateral, and sequential manner in which trades are executed should provide a more realistic basis for calculating potential cost savings. This paper presents empirical evidence on potential cost savings by examining a market for the abatement of sediment from farmland. Empirical results based on a market simulation model find no statistically significant change in mean abatement costs under several transaction cost levels when contracts are randomly executed. An alternative method of contract execution, gain-ranked, yields similar results. At the highest transaction cost level studied, trading reduces the total cost of compliance relative to a uniform standard that reflects current regulations. PMID- 11381953 TI - The influence of psychological stress on periodontal disease. AB - This paper reviews studies that investigated the association between psychosocial stress and periodontal disease. The impact of stress on the immune system has been well established, and a possible influence on a chronic inflammatory disease like periodontitis appears probable. Yet, a direct association between periodontal disease and stress remains to be proven, which is partly due to lack of an adequate animal model and the difficulty in quantifying amount and duration of stress and the detrimental effects of inadequate coping with stress. Multiple variables affecting the severity of periodontal disease and the uncertainty about the individual onset of periodontal disease complicate the issue further. Nevertheless, more recent studies indicate that psychosocial stress represents a risk indicator for periodontal disease and should be addressed before and during treatment. PMID- 11381954 TI - Relevance of microbial extracellular polymeric substances (EPSs)--Part I: Structural and ecological aspects. AB - Extracellular polymeric substances are the construction materials for microbial aggregates such as biofilms, flocs ("planktonic biofilms") and sludge. Their major components are not only polysaccharides but also proteins and in some cases lipids, with minor contents of nucleic acids and other biopolymers. In the EPS, biofilm organisms can establish stable arrangements and function multicellularly as synergistic microconsortia. The matrix facilitates the retention of exoenzymes, cellular debris and genetic material; it can be considered as a microbial recycling yard. Gradients can develop due to the physiological activity and the fact that diffusive mass transport prevails over convective transport in the matrix. Biofilm cells tolerate higher concentrations of many biocides. The EPS matrix sequesters nutrients from the water phase. In photosynthetic communities, EPS molecules can function as light transmitters and provide photons to organisms located deeper in a microbial mat. The EPS matrix is a dynamic system, constructed by the organisms and responding to environmental changes. It enables the cells to function in a manner similar to multicellular organisms. PMID- 11381955 TI - SEM-EDS for determining the phosphorus content in activated sludge EPS. AB - Not all phosphorus removed in activated sludge systems can be accounted for by polyphosphate accumulating organisms (PAO). A method for the qualitative and quantitative in situ characterization of PAO cell clusters and closely associated extracellular polysaccharides (EPS) is described. X-ray microanalysis was performed on samples from four activated sludge plants situated in Pretoria, South Africa. Analyses were done by means of Scanning Electron Microscopy (SEM) combined with Energy Dispersive Spectrometry (EDS). Cell clusters with associated EPS on average contained between 57 and 59% phosphorus, while EPS alone contained on average between 23 and 30% phosphorus. Results suggest that phosphorus removal in activated sludge might be due not only to PAO, but also by EPS acting as a phosphorus reservoir. Extraction of EPS from two different activated sludge plants yielded different amounts of EPS, which, in combination with SEM-EDS, may shed light on different phosphate uptake abilities of different activated sludges. PMID- 11381956 TI - The influence of fluid shear and AICI3 on the material properties of Pseudomonas aeruginosa PAO1 and Desulfovibrio sp. EX265 biofilms. AB - An understanding of the material properties of biofilms is important when describing how biofilms physically interact with their environment. In this study, aerobic biofilms of Pseudomonas aeruginosa PAO1 and anaerobic sulfate reducing bacteria (SRB) biofilms of Desulfovibrio sp. EX265 were grown under different fluid shear stresses (tau g) in a chemostat recycle loop. Individual biofilm microcolonies were deformed by varying the fluid wall shear stress (tau w). The deformation was quantified in terms of strain (epsilon), and the relative strength of the biofilms was assessed using an apparent elastic coefficient (Eapp) and residual strain (epsilon r) after three cycles of deformation. Aluminium chloride (AlCl3) was then added to both sets of biofilm and the tests repeated. Biofilms grown under higher shear were more rigid and had a greater yield shear stress than those grown under lower shear. The addition of AlCl3 resulted in a significant increase in Eapp and also increased the yield point. We conclude that the strength of the biofilm is in part dependent on the shear under which the biofilm was grown and that the material properties of the biofilm may be manipulated through cation cross-linking of the extracellular polysaccharide (EPS) slime matrix. PMID- 11381957 TI - Modelling the structure and function of extracellular polymeric substances in biofilms with new numerical techniques. AB - The objective of this study is the mathematical description of the structure and function of the extracellular polymeric substances (EPS) in biofilms. The basic assumptions of the EPS biofilm model are: the production of EPS in biofilms is coupled to the growth of micro-organisms the production of EPS is additionally coupled to the substrate conditions the EPS represent a considerable volume fraction of the matrix in biofilms and thus the density of the biofilms is strongly influenced by the EPS sorption of biocides and pollutants in biofilms occurs mainly to EPS the EPS can be used as an energy source during substrate limited phases. The mathematical model has been derived as a system of partial differential equations. The numerical solution of these complex balance equations has been done by a self-adaptive Galerkin-h-p-method. It can be shown, that on the one hand the simulation of substrate conversion and biofilm growth with the EPS-biofilm model yields similar results as the known biofilm models without consideration of the EPS fraction. On the other hand the advantage of the EPS biofilm model is a better understanding of biofilm structure, which is mainly influenced by the EPS fraction in the biofilm. Furthermore, the sorption of pollutants, such as heavy metals and chlorinated organic substances, can be simulated in more detail. PMID- 11381958 TI - Modelling production of extracellular polymeric substances in a Pseudomonas aeruginosa chemostat culture. AB - Formation of extracellular polymeric substances (EPSs) by mucoid Pseudomonas aeruginosa was investigated using literature data from chemostat cultures. The data were used to calibrate a product formation regime allowing substrate sufficient and endogenous EPS formation. Yield coefficients for both formation conditions were elucidated based on metabolic pathway analysis. Growth and non growth related specific formation rates of 0.18 g CEPS/g Ccell/h and 0.03 1/h were estimated, respectively. The exogenous and endogenous EPS yield was found to be 0.77 g CEPS/g Cglu and 0.79 g CEPS/g Ccell, respectively. Being structurally equivalent to comprehensive maintenance models, this model allows for non-growth related product formation, showing the same quality of fit as previous models restricted to exogenous EPS precursors. PMID- 11381959 TI - Effect of EPS on biofilm structure and function as revealed by an individual based model of biofilm growth. AB - We have simulated a nitrifying biofilm with one ammonia and one nitrite oxidising species in order to elucidate the effect of various extracellular polymeric substance (EPS) production scenarios on biofilm structure and function. The individual-based model (IbM) BacSim simulates diffusion of all substrates on a two-dimensional lattice. Each bacterium is individually simulated as a sphere of given size in a continuous, three-dimensional space. EPS production kinetics was described by a growth rate dependent and an independent term (Leudeking-Piret equation). The structure of the biofilm was dramatically influenced by EPS production or capsule formation. EPS production decreased growth of producers and stimulated growth of non-producers because of the energy cost involved. For the same reason, EPS accumulation can fall as its rate of production increases. The patchiness and roughness of the biofilm decreased and the porosity increased due to EPS production. EPS density was maximal in the middle of the vertical profile. Introduction of binding forces between like cells increased clustering. PMID- 11381960 TI - Time-resolved study of biofilm architecture and transport processes using experimental and simulation techniques: the role of EPS. AB - Cellular material and extracellular polymeric substances are the basic structural elements in biofilm systems. The structure and role of EPS for biofilm development and metabolic processes have not been precisely determined and, therefore, have not yet been included as a necessary element in modelling and simulation studies. This is due to the difficulty of experimentally detecting the extracellular polymeric substances in situ and differentiating them from cellular material on the one hand, and to the subsequent uncertainty about appropriate models--e.g. rigid hindrances, porous microstructure or visco-elastic structure- on the other hand. In this work, we report on the use of confocal laser scanning microscopy to monitor the development of a monoculture biofilm of Sphingomonas sp. grown in a flow cell. The bacterial strain was genetically labelled resulting in strong constitutive expression of the green fluorescent protein. The development of extracellular polymeric substances was followed by binding of the lectin concavalin A to cell exopolysaccharides. The growth of the resulting strain was digitally recorded by automated confocal laser scanning microscopy. In addition, local velocity profiles of fluorescent carboxylate-modified microspheres were observed on pathlines throughout the biofilm. The CLSM image stacks were used as direct input for the explicit modelling and three-dimensional numerical simulation of flow fields and solute transport processes based on the conservation laws of continuum mechanics. At present, a strongly simplifying EPS model is applied for numerical simulations. The EPSs are preliminarily assumed to behave like a rigid and dense hindrance with diffusive-reactive solute transport. PMID- 11381961 TI - Methanotrophic production of extracellular polysaccharide in landfill cover soils. AB - A bench-scale soil reactor was used to study methane oxidation and EPS production under tropical conditions. The study of pertinent environmental factors affecting EPS production was carried out by batch cultivation of methanotrophs. These factors included variations in temperature (20 degrees C to 45 degrees C), soil water content (5% to 33%), and the supply ratios of methane/oxygen. The bench scale study revealed that excessive EPS was accumulating in an active methane oxidation zone located 5-45 cm below the soil surface of the reactor. The observed peak rates of oxidation could not be sustained over an extended period of time due to EPS accumulation. Results from the batch cultivation experiments confirmed the production of EPS in soils subject to methane oxidation. EPS production was found to correlate with methane oxidation rates which, in turn, were regulated by the variance of temperature and soil water content. A larger amount of EPS production was obtained at 30 degrees C and 17% soil water content. Oxygen is required for methane oxidation; however, at high oxygen tension it may accelerate the production of EPS by methanotrophs causing limited oxygen diffusion and declining rates of methane oxidation. PMID- 11381962 TI - The EPS of Acidithiobacillus ferrooxidans--a model for structure-function relationships of attached bacteria and their physiology. AB - To dissolve pyrite or sulphur, leaching bacteria like Acidithiobacillus ferrooxidans attach to these substrata by extracellular polymeric substances (specifically, lipopolysaccharides). The primary attachment to pyrite at pH 2 is mediated by exopolymer-complexed iron(III) ions in an electrostatic interaction with the negatively charged pyrite surface. Cells grown on sulphur exhibit a different composition of the extracellular lipopolysaccharides, namely with increased hydrophobic properties, and do not attach to pyrite. Thus, the cells adapt the chemical composition of their exopolymers to the substrate/substratum. It is concluded that the mechanism of bacterial pyrite oxidation is basically indirect. The actual corrosive agents are iron(III) ions. Preliminary data indicate that active strains complex more iron(III) ions in their EPS than less active ones. Obviously, the exopolymeric layer comprises a reaction space for the regeneration of these ions by the activity of the iron oxidising bacteria. PMID- 11381963 TI - Agarose hydrogels as EPS models. AB - For investigating the influence of extracellular polymeric substances (EPSs) on biofilm properties, artificial models of defined composition and structure can be very helpful. Different immobilised cell systems can be useful in the fitting of experimental results. Two different types of of artificial EPS matrix model were developed earlier. Homogeneous agarose beads (50-500 microns diameter) and porous beads (260 microns mean diameter) containing pores with diameters from 10 to 80 microns (28 microns on average), allowed the embedding of cells, particles and typical EPS matrix components such as proteins and polysaccharides. In this paper, some physico-chemical properties of the artificial EPS matrices were described and compared with results known from natural EPS. The stability of the artificial matrix polymer against solution in the aqueous medium was studied in batch experiments. The water binding and water retaining capabilities of the EPS models were investigated by drying and re-swelling experiments. The simulation of protective effects of the artificial EPS matrix against toxic substances like biocides in comparison to such known protective effects of the EPS of native biofilms were proved by the application of sodium hypochlorite (0.5 mg/l, 30 min) and subsequent microscopic investigation of the cell population after LIVE/DEAD staining (Molecular Probes). PMID- 11381964 TI - Remember the water--a comment on EPS colligative properties. AB - The relationship between water and activated sludge components was examined. Reevaluation of published data on freezing point depression, drying rates and dewatering has been performed. The basis of this has been the assumption that the water/sludge relationship is considered to be a colligative effect. Since the results indicate this to be the case, we suggest that the published concepts of "pools of water" are false. Data on swelling properties of EPS as a function of pH suggests that the colligative properties are largely determined by the counterions of charged polymers and surfaces. PMID- 11381965 TI - Cell surface and exopolymer characterization of laboratory stabilized activated sludge from a beverage bottling plant. AB - Fermentor-stabilized activated sludge from an industrial beverage bottling plant was grown on three different food sources: normal plant wastewater, plant wastewater containing high sucrose concentrations, and a synthetic glucose-based feed stock. Surface charge, hydrophobicity, and exopolysaccharide composition were measured on the stabilized bacterial flocs. Cell surface charge was measured by electrophoretic mobility, dye exchange titration, and a standard colloid titration, while cell hydrophobicity was determined using the bacterial adhesion to hydrocarbons (BATH) test. Exopolysaccharide profiles were determined by measuring concentrations of glucose, galactose, mannose, glucuronic, and galacturonic acids in digested exopolymer extractions using HPLC. Changes in the physical surface properties of the bacteria and the chemical composition of the extracted exopolymers were correlated with differences in the three food sources. Cell surface hydrophobicity was similar for cultures grown on different plant wastewaters, while the culture grown on synthetic food produced less floc hydrophobicity. Electrophoretic mobility measurements, charge titrations, and dye exchange titrations showed different total surface charge as well as varying charge availability. Additionally, total surface charge and total exopolysaccharide concentrations appeared less dependent on food source than the food-to-mass ratio. High concentrations of biodegradable food produced dispersed growth and high concentrations of exopolysaccharides that contributed to poor settling. PMID- 11381967 TI - Effect of carbohydrate and protein in the EPS on sludge settling characteristics. AB - EPSs have been believed to play a bonding role in microbial floc formation. However, the precise role is not well known. In this study, sludge settling characteristics and the carbohydrate to protein ratio in the EPS were tested at various airflow rates. Sludge was collected from three modified sequencing batch reacetors (MSBRs), which were operated with airflow rates of 0.8 L/min, 2 L/min and 4 L/min, respectively. During the operation periods, the reactor operated at an airflow rate of 0.8 L/min showed a sludge volume index (SVI) of 80 to 90 mL/g and a constant ratio of carbohydrate to protein in the EPS, while a significant increase of this ratio and the SVI occurred in the other reactors. High airflow rate increased the amount of carbohydrate in the EPS, but the protein level was almost constant for reactors with airflow rates of 2 L/min and 4 L/min. The higher ratio of carbohydrate to protein caused the bulking of the sludge; hence it was not favourable for sludge settling. The ratio of carbohydrate and protein in the EPS is inferred to be essential for solid floc formation. PMID- 11381966 TI - Extracellular polymeric substances in relation to nutrient removal from a sequencing batch biofilm reactor. AB - Experimental investigations were performed to determine the possibility of simultaneous biological nitrogen and phosphorus removal during various biofilm processes in conjunction with biofilm characterisation, especially extracellular polymeric substance (EPS). Since biologial nitrogen removal requires an alternating exposure of anaerobic-anoxic-oxic conditions in the bulk liquid that surrounds the biofilm growth media, a sequencing batch reactor (SBR)-type operation was used. Various materials including expanded clay, polystyrene, polyurethane, and acrylic materials were used as the biofilm growth support medium. Simultaneous nitrogen and phosphorus removal was possible with SBR, but it was postulated that nutrient removal efficiencies varied with film thickness. Thinner biofilm promoted nitrification and phosphorus removal, but thicker biofilm enhanced denitrification and reduced phosphorus removal. EPS contents were similar regardless of support media types or biofilm configuration, but EPS contents gradually increased as the film growth continued after backwashing. EPS contents were increased with increased nitrogen removal, but it was difficult to define its relation with phosphorus removal. In addition, suspended solids removal was correlated well with the EPS content in the biofilms. PMID- 11381968 TI - Enhancement of nitrifying biofilm formation using selected EPS produced by heterotrophic bacteria. AB - The possibility of enhancing nitrifying biofilm formation rate with the aid of selected EPS produced by heterotrophic bacteria was investigated. When EPS production characteristics were examined for four kinds of heterotrophs isolated from a domestic wastewater treatment reactor, two strains obtained from biofilms (B1, B2) exhibited a higher polysaccharide production rate than those from suspended flocs (A1, A2). Among EPS components, the concentration of uronic acids gave a good correlation with flocculation ability, which suggests that acidic polysaccharides play a major role in bioaggregate formation. Addition of 1 g/L D glucuronic acid as an EPS substitute enhanced the homocoagulation rate of autotrophic Nitrosomonas europaea and altered its zeta potential from n30.4 mV to +4.3 mV, which indicates a possibility that particular EPS components produced by heterotrophs are utilized as neutralising reagents for nitrifying biofilm formation. Moreover, when heterotrophic isolates with Nitrobacter winogradskyi were cultured in batch with fabric supports, biofilm formed on the substratum. These experimental results suggest the application of selected EPS for enhancing nitrifying biofilm formation. PMID- 11381969 TI - A new method for extraction of extracellular polymeric substances from biofilms and activated sludge suitable for direct quantification of sorbed metals. AB - A method for extraction of extracellular polymeric substances (EPSs) with a dicyclohexyl-18-crown-6 ether was developed to determine levels of organic and inorganic contaminants sorbed to EPS. The crown ether selectively binds alkaline and alkaline earth metals but not heavy metals. The effectiveness of the extraction procedure was higher than that of 2 other methods tested and comparable with that of a method based on a cation exchange resin. On average it was possible to extract 20% of the TOC, 12% of the total protein content, and 4% of the total carbohydrate content of sludge or biofilm biomass. Metal sorption studies in activated sludge showed no influence of exposure time on the fractionation of metals within the biomass. Metals sorbed mostly to cellular material. In biofilms 12.2% of the cadmium and 9.1% of the zinc added was found in the EPS. In activated sludge EPS contained only 2.9% zinc. The distribution of metals within the biomass was dose dependent. The percentage of metals found in EPS decreased with increasing metal concentration. This indicates a higher affinity of metals for cellular binding sites. Time course experiments in a rotating biofilm annular reactor, which consisted of an external cylinder with removable slides and an internal solid drum, revealed a gradual change in zinc concentration associated with EPS, although the total zinc concentration in the biomass remained constant. Concurrently, the amount of extractable EPS decreased. This was a consequence of a microbial population shift, with bacterial counts decreasing and algal and fungal biomass increasing. Using confocal laser scanning microscopy and the fluorescent metal complexing agent Newport Green for in situ detection of zinc it was shown that metals were bound to algae and fungi in the latter part of the experiment. The biofilm became more and more heterogeneous coinciding with a decrease in EPS. To summarize, the observed sorption behavior of metals cannot be explained with the conventional paradigm of EPS as hydrophilic gel. Obviously, different binding mechanisms must be invoked to explain the role of EPS in the sorption and removal of toxic substances in activated sludge and biofilm systems. It is important to consider the microbial population to understand differences in sorption in different matrices. PMID- 11381970 TI - Hydrolysis of wastewater colloidal organic matter by extracellular enzymes extracted from activated sludge flocs. AB - Enzymatic activities associated with the exopolymeric substances (EPSs) extracted from activated sludges were tested for their ability to hydrolyse the organic colloidal fraction of wastewater. Bacteria extracted with EPS and concentrated by wastewater microfiltration were inhibited with NaN3 or KCN. The protein hydrolysis mainly resulted from the enzymatic activity of EPS, whereas the glycolytic activity was mainly present in the organic colloidal fraction of the wastewater. PMID- 11381971 TI - Effect of heavy metals (Cu, Pb, and Ni) on the compositions of EPS in biofilms. AB - A series of batch adsorption tests to estimate the biofilm capacity for removal of the heavy metals (Cu, Pb, and Ni) in wastewater were carried out. The metal sorption results were fitted to the Freundlich isotherm model to compare their sorption capacity. The change of the composition of EPSs (extracellular polymeric substances) was simultaneously represented by the ratio of carbohydrate to protein (C/P) when the biofilm was exposed to the heavy metals. EPS composed of slime loosely bound to the cell and capsular materials was extracted by the four general extraction methods including regular centrifugation, regular centrifugation with formaldehyde, EDTA extraction and steam extraction. Although the various extraction methods showed different results, C/P ratio of biofilm exposed to copper and lead metal ions was generally lower than that of control. PMID- 11381972 TI - Influence of calcium ions on the mechanical properties of a model biofilm of mucoid Pseudomonas aeruginosa. AB - The mechanical properties of biofilms and in particular their mechanical strength is of great importance for both biofilm reactors and for the removal of undesired biofilms as in cases of biofouling and biocorrosion. By uniaxial compression measurements, it is possible to determine the apparent elastic or Young's modulus and the yield stress as parameters for mechanical stability. This was performed with a recently developed device, using model biofilms of mucoid strain Pseudomonas aeruginosa SG81. The biofilms were grown on membrane filters placed on nutrient agar medium with different concentrations of calcium ions. The compressive stress-strain behaviour up to failure was recorded at a compression speed of 1 micron s-1. The apparent Young's modulus, representing the stiffness of the biofilm, and the yield stress obtained from the stress--strain diagram were used for the description of mechanical properties of biofilms. A certain critical concentration of calcium ions was found where the Young's modulus of the P. aeruginosa biofilms increases strongly and subsequently remains constant for higher calcium concentrations. This behaviour is explained by the presence of calcium ions crosslinking alginate, which is the major component of the extracellular polymeric substances produced by the mucoid P. aeruginosa strain used in this investigation. PMID- 11381973 TI - Adsorption of heavy metals by EPS of activated sludge. AB - Extracellular polymeric substances (EPSs) were extracted by high-speed centrifugation at 20,000 G for 30 min from an activated sludge treating municipal wastewater. Each gram of sludge, as measured in volatile suspended solids (VSS), contained 7.3 mg of EPS, including 6.5 mg of protein (EPSp) and 0.8 mg of carbohydrate (EPSc). The EPSp had a mean MW of 2.0 x 105; about 18% of EPSp had MW over 5 x 104 and 16% below 5 x 103. For heavy metal concentrations ranging 10 100 mg/l, EPS on average removed 99% of Zn2+, 98% of Cu2+, 97% of Cr3+, 85% of Cd2+, 69% of Co2+, 37% of Ni2+, and 26% of CrO4(2-). The relative degrees of metal removals were inconsistent with those reported for the activated sludge process. Each mg of ESP had the capacity to remove up to 1.48 mg of Zn2+, 1.12 mg of Cu2+, 0.83 mg of Cr3+, 0.90 mg of Cd2+, 1.10 mg of Co2+, 0.25 mg each of Ni2+ and CrO4(2-). Results suggest the feasibility of recovering ESP from waste sludge for use as adsorbent. Freundlich isotherm correlated satisfactorily with the adsorption data of Ni2+, Cu2+, Cd2+, and CrO4(2-) (R2 ranging 0.89-0.97), whereas Langmuir isotherm correlated satisfactorily with those of Zn2+, Cr3+ and Ni2+ (R2 ranging 0.93-0.96). Both correlated poorly for those of Co2+. PMID- 11381974 TI - Quantification of the bond energy of bacteria attached to activated sludge floc surfaces. AB - The great majority of activated sludge bacteria exist incorporated in flocs. The increase in dispersed bacteria when exposed to increasing turbulent shear rates has been successfully modelled by a model assuming that the adhesion and erosion of cells may be considered in analogy to ordinary chemical phase transitions. By this adhesion-erosion model (AE-model), an "enthalpy" of cell adhesion can be estimated, and this value in turn determines the range of shear rates in which erosion of cells predominates. Application of the model has indicated that only a mass fraction less than ca. 6-17% may be released from activated sludge, even when exposed to a severe turbulent environment, i.e. only a small fraction of the flocs is dispersible by means of erosion by turbulence. The shear sensitivity and the dispersible floc fraction were found to depend on the floc composition. A net decrease in the floc EPS content during anaerobic sludge stabilisation causes a dramatic increase in the dispersed fraction, indicating the important role of EPS for the floc strength. It was found also that activated sludge cells do not reflocculate completely after exposure to high shear rates. This may be an indication that the cohesion energy of bacteria growing in colonies is greater than the energy of the more stochastic adhesion of dispersed cells to floc surfaces. It could also be another indicator of the importance of entanglement forces, which do not reform instantly, once broken. When the bond strength of cell attachment to sludge surfaces is altered by changes in the chemical environment, this may cause a change in the dispersible floc fraction as well as a change in the shear range of erosion. When the shear sensitivity constant kss is adopted for sludge characterisation, an increased degree of dispersion under standard test conditions will result in increased shear sensitivity estimates. The shear sensitivity may be used for the estimation of en equivalent change in the Gibb's energy of cell adhesion (delta (delta Gad/RT)). PMID- 11381975 TI - Exopolysaccharides in biofilms, flocs and related structures. AB - In biofilms, flocs and similar multispecies microbial communities, exopolysaccharides (EPSs) are always present, frequently as the major component other than water. The EPSs vary widely in their composition, structure and properties and thus it is impossible to generalise about their contribution to biofilm or floc structure. Relatively few of the polymers obtained from biofilms and flocs have been adequately purified and analysed but such evidence as is so far available suggests that the polysaccharides closely resemble those synthesised by the corresponding planktonic bacteria. From a knowledge of the physical properties of these, it is now possible to present a reasonably accurate picture of some of the factors which they may contribute to the structure and stability of complex aggregates of micro-organisms in biofilms and flocs. PMID- 11381976 TI - Fluorescence spectroscopy and molecular weight distribution of extracellular polymers from full-scale activated sludge biomass. AB - Two fractions of extracellular polymer substances (EPSs), soluble and readily extractable (RE), were characterised in terms of their molecular weight distributions (MWD) and 3-D excitation-emission-matrix (EEM) fluorescence spectroscopy signatures. The EPS fractions were different: the soluble EPSs were composed mainly of high molecular weight compounds, while the RE EPSs were composed of small molecular weight compounds. Contrary to previous thought, EPS may not be considered only as macromolecular because most organic matter present in both fractions had low molecular weight. Three different fluorophore peaks were identified in the EEM fluorescence spectra. Two peaks were attributed to protein-like fluorophores, and the third to a humic-like fluorophore. Fluorescence signatures were different from other previously published signatures for marine and riverine environments. EEM spectroscopy proved to be a suitable method that may be used to characterise and trace organic matter of bacterial origin in wastewater treatment operations. PMID- 11381977 TI - Relevance of microbial extracellular polymeric substances (EPSs)--Part II: Technical aspects. AB - Extracellular polymeric substances (EPSs) are involved in both detrimental and beneficial consequences of microbial aggregates such as biofilms, flocs and biological sludges. In biofouling, they are responsible for the increase of friction resistance, change of surface properties such as hydrophobicity, roughness, colour, etc. In biocorrosion of metals they are involved by their ability to bind metal ions. In bioweathering, they contribute by their complexing properties to the dissolution of minerals. The EPSs represent a sorption site for pollutants such as heavy metal ions and organic molecules. This can lead to a burden in wastewater sludge; on the other hand, the sorption properties can be used for water purification. Other biotechnological uses of EPS exploit their contribution to viscosity, e.g., in food, paints and oil-drilling 'muds'; their hydrating properties are also used in cosmetics and pharmaceuticals. Furthermore, EPSs may have potential uses as biosurfactants, e.g., in tertiary oil production, and as biological glue. EPSs are an interesting component of all biofilm systems and still hold a large biotechnological potential. PMID- 11381978 TI - In situ detection of cell surface hydrophobicity of probe-defined bacteria in activated sludge. AB - The surface hydrophobicity of different types of bacteria in activated sludge were investigated under in situ conditions by following the adhesion of fluorescent microspheres with defined surface properties to bacterial surfaces (the MAC-method). This technique was combined with identification of the bacteria with fluorescence in situ hybridization with rRNA-targeted oligonucleotides (FISH) and could thus be used for characterization of surface properties of probe defined bacteria directly in a complex system without prior enrichment or isolation. This MAC-FISH technique could be used for single bacteria as well as filamentous bacteria. In the investigated activated sludge from an industrial wastewater treatment plant, two types of filamentous bacteria dominated. One morphotype consistently attracted only very few hydrophobic microspheres, indicating that the thin sheath of exopolymers around the cells had a hydrophilic surface. Use of a hierarchical set of gene probes revealed that these filaments were sulphide oxidising Thiothrix spp. The other predominating filamentous morphotype had a thick, very hydrophobic exopolymeric sheath. This filamentous bacterium was found to belong to the alpha-Proteobacteria. The relevance of the significant differences in surface hydrophobicity for the two morphotypes in respect to substrate uptake and floc formation is discussed. PMID- 11381979 TI - Monitoring of a biogas process using electronic gas sensors and near-infrared spectroscopy (NIR). AB - The use of electronic gas sensors and near-infrared spectroscopy (NIR) to monitor the dynamics in a biogas process was evaluated using multivariate data analysis. The digester, a completely stirred 8 l tank reactor fed with a mixture of cellulose, albumin and minerals, was exposed to an overload of glucose after which monitoring of electronic gas sensor responses, NIR spectra as well as traditional chemical variables and analysis of microbial community structure were done. The responses from an array of electronic gas sensors consisting of MOS and MOSFET-sensors were correlated against volatile compounds in the headspace using partial least square (PLS) regressions. The root mean square error of prediction (RMSEP) was 0.15 g/l for acetate in the range of 0.14-1.72 g/l and the RMSEP for methane was 2.3% in the range of 27-73%. Selected wavelengths from the second derivative of the original NIR spectra (400-2500 nm) resulted in a PLS-model for predicting microbial biomass, measured as total phospholipid fatty acids, with a RMSEP of 9 nmol/ml in the range of 163-293 nmol/ml. The NIR model developed for acetate had a RMSEP of 0.20 g/l within the range of 0.14-1.72 g/l. The results clearly show that both NIR and an array of electronic gas sensors can provide simultaneous non-invasive in situ monitoring of important process variables in anaerobic digesters. PMID- 11381980 TI - Pilot-scale gasification of municipal solid wastes by high-rate and two-phase anaerobic digestion (TPAD). AB - Bioconversion of municipal solid waste-sludge blend by conventional high-rate and two-phase anaerobic digestion was studied. RDF (refused-derived fuel)-quality feed produced in a Madison, Wisconsin, USA, MRF (materials-recovery facility) was used. High-rate digestion experiments were conducted with bench-scale digesters under target operating conditions developed from an economic feasibility study. The effects of digestion temperature, RDF content of digester feed, HRT, loading rate, RDF particle size, and RDF pretreatment with cellulase or dilute solutions of NaOH or lime on digester performance were studied. A pilot-scale two-phase digestion plant was operated with 80:20 (weight ratio) RDF-sludge blends to show that this process exhibited a higher methane yield, and produced a higher methane content digester gas than those obtained by single-stage, high-rate anaerobic digestion. PMID- 11381981 TI - Two-phase anaerobic digestion of source sorted OFMSW (organic fraction of municipal solid waste): performance and kinetic study. AB - The results of a two-phase system operated in different conditions, treating the source-sorted organic fraction of municipal solid waste (SS-OFMSW), coming mainly from fruit and vegetable markets, are presented. Hydraulic retention time (HRT) in the hydrolytic reactor and in the methanogenic reactor and also the temperature in the hydrolytic reactor (mesophilic and thermophilic conditions) are varied in order to evaluate the effect of these factors. The methanogenic reactor is always operated within the thermophilic range. Optimum operating conditions are found to be around 12 days (total system) using the mesophilic range of temperature in the first reactor. Specific gas production (SGP) in these conditions is around 0.6 m3/kg TVS. A kinetic study is also carried out, using the first and the step diffusional models. The latter gives much better results, with fitted constants comparable to other studies. Finally, a comparison with a one-phase system is carried out, showing that a two-phase system is much more appropriate for the digestion of this kind of highly biodegradable substrate in thermophilic conditions. PMID- 11381982 TI - Effect of moisture content on anaerobic digestion of dewatered sludge: ammonia inhibition to carbohydrate removal and methane production. AB - The purpose of this study is to investigate the effect of moisture content on anaerobic digestion of dewatered sewage sludge under mesophilic condition. The moisture contents of sludge fed to reactors were 97.0%, 94.6%, 92.9%, 91.1% and 89.0%. The VS removal efficiency changed from 45.6% to 33.8%, as the moisture content of sludge fed to digester decreased from 97.0% to 89.0%. The carbohydrate removal efficiency also decreased from 71.1% to 27.8%. Methane production decreased when the moisture content of sludge was lower than 91.1%. The number of glucose consuming acidogenic bacteria was decreased from 3.1 x 10(6) to 3.1 x 10(8) (MPN/mL) as the moisture content decreased from 91.1% to 89.0%. The numbers of hydrogenotrophic and acetoclastic methanogenic bacteria decreased by one order of magnitude when the moisture content was lower than 91.1%. The decrease in numbers of glucose consuming acidogenic bacteria and methanogenic bacteria was found to correspond to the decrease in the carbohydrate removal efficiency and the accumulation of propionic acid. Batch experiments showed that acetoclastic methanogenic bacteria were acclimated to high ammonia concentration, on the other hand, glucose consuming acidogenic bacteria were inhibited. PMID- 11381983 TI - Fact-finding survey of actual garbage discharged from dormitory and its biological anaerobic-aerobic treatment. AB - The objective of this study is to find a possibility of complete treatment of garbage and resource recovery (production of methane from available utility of carbon resource in garbage) by biological treatment process. As the first step, a fact-finding survey of actual garbage discharged from the dormitory of the Ube National College of Technology (equivalent to 300 population) was carried out. Second, the combined biological anaerobic-aerobic treatment, i.e. combination of upflow anaerobic sludge blanket (UASB) process and aerobic membrane bioreactor (AMB) process, was applied to the garbage treatment. The applicability and efficiency of this system were investigated in this study. The survey results showed that the composition and quantity of garbage from a student dormitory changed slightly during a week due to the change of the menu, however, they remained almost unchanged during the entire experimental period. The experimental results showed high biodegradability of the garbage, and demonstrated its suitability for methane production. The soluble nitrogen removal was high: over 97%. No excess sludge was wasted from the system. A high treatment efficiency of simultaneous organic carbon and nitrogen was obtained. The possibility of complete treatment of garbage with this process has been positively demonstrated by this study. PMID- 11381984 TI - Investigations on the changes in anaerobic biodegradability and biotoxicity of an industrial microbial biomass induced by a thermochemical pretreatment. AB - In the anaerobic digestion of solid wastes, hydrolysis is the rate limiting step and physicochemical pretreatment is often required to promote solubilization of organic matter. As an illustration, anaerobic digestion was limited by the substrate solubilization step during continuous cultures performed with an industrial microbial biomass. In optimal operating conditions determined for the hydrolysis-acidogenesis step (T = 35 degrees C; pH = 8.5; OLR = 5.4 g COD/l.d) 50.6% COD solubilization was achieved. A thermochemical pretreatment based on sodium hydroxide addition, was used in order to enhance COD solubilization. Optimal conditions for COD solubilization were pH = 12, T = 140 degrees C for 30 minutes. In these conditions, 70% COD solubilization was achieved. However, anaerobic biodegradability of the pretreated substrate was not improved and remained near 40%. The poor anaerobic biodegradability performances were attributed to the soluble molecules generated during the thermochemical pretreatment that were refractory and/or inhibitory to anaerobic microorganisms. Fractionation of the soluble pretreated microbial biomass by two methods (treatment with adsorbent resins and precipitation by pH adjustment) demonstrated that high molecular weight compounds (> 100 kDa) are involved in the poor biodegradability and in the biotoxicity observed. Partial decolorization through resin use and acid precipitation remove these compounds. The consequence of their removal was an increase of the production of biogas. PMID- 11381985 TI - Increase of anaerobic degradation of particulate organic matter in full-scale biogas plants by mechanical maceration. AB - Different concepts of implementation of mechanical pretreatment for enhancing the biogas potential from fibers in manure feedstock were evaluated by sampling before and after macerators at different biogas plants and from a fiber separation unit. An increase of the biogas potential of up to 25% by pretreatment of the whole feed in the macerator before the reactor was observed. Implementation concepts with a treatment of the fibers alone after separation from the manure showed to be not efficient due to a low recovery of organic matter in the fibers by the separation unit. The low operational costs of a macerator make it attractive to use this pretreatment method for a more complete degradation of particulate organic matter. Investigation of the size distribution of the fibers showed that a change in biogas potential was not correlated to a smaller size of the fibers. Results from the macerators indicate that the biodegradability of the fibers is rather enhanced by shearing which is not necessarily reflected by a change in fiber size. PMID- 11381986 TI - Influence of the size reduction of organic waste on their anaerobic digestion. AB - The rate-limiting step in anaerobic digestion of organic solid waste is generally their hydrolysis. A size reduction of the particles and the resulting enlargement of the available specific surface can support the biological process in two ways. Firstly, in case of substrates with a high content of fibres and a low xegradability, their comminution yields to an improved digester gas production. This leads to a decreased amount of residues to be disposed of and to an increased quantity of useful digester gas. The second effect of the particle size reduction observed with all the substrates but particularly with those of low degradability is a reduction of the technical digestion time. Furthermore, the particle size of organic waste has an influence on the dewaterability after codigestion with sewage sludge. The presence of organic waste residues improves the dewaterability measured as specific resistance to filtration but this positive effect is attenuated if the particle size of the solids is reduced. PMID- 11381987 TI - Solubilization of organic sludge by thermophilic aerobic bacteria as a pretreatment for anaerobic digestion. AB - Solubilization of organic sludge by thermophilic aerobic bacteria as a pretreatment for anaerobic digestion was investigated. The thermophilic bacteria which solubilized organic sludge were isolated from the thermophilic aerobic digestion reactor. The bacterium type SPT2-1 could grow at pH ranging from 5.0 to 8.5 with optimal temperature at 60-70 degrees C. In batch experiments, 25-30% of volatile suspended solids (VSS) in the pre-heated sludge were solubilized on inoculating with the isolated bacteria although little was solubilized without inoculation. The isolated bacteria appeared to secret the extracellular enzymes including proteases and amylases. In continuous flow experiments, sludge solubilization rate (VSS removal) was around 40% under aerobic as well as microaerobic conditions. No accumulation of volatile fatty acids in the treated sludge was observed under aerobic conditions while significant amounts of them were accumulated under microaerobic conditions. Production of biogas on anaerobic digestion of the microaerobically-pretreated sludge was increased by 1.5 when compared with the sludge without pretreatment. PMID- 11381988 TI - Anaerobic hydrolysis kinetics of particulate substrates. AB - A mathematical description of the surface related hydrolysis kinetics for spherical particles in a batch digestion is presented as well as a verification of this model with particulate starch as a substrate. Three substrates containing starch with different particle size distributions (PSD) were used. Two were obtained from fresh potatoes by wet sieving and for the third substrate a commercially available starch was used. The substrates were batch digested at 30 degrees C with granular sludge as inoculum and the hydrolysis efficiency was measured and fitted with the model. The results revealed that the hydrolysis rates for the three substrates were equal, viz. 0.4 +/- 0.1 g starch/m2/hour. Moreover, for the commercial starch not only the hydrolysis efficiency but also the changes within the PSD of the starch was determined several times with the use of light microscopy and image analysis. The obtained experimental PSD showed good similarity with the theoretical PSD from model calculations. This shows that the surface of the particulate substrate is the key factor for the hydrolysis process. PMID- 11381989 TI - Effects of disintegration on anaerobic degradation of sewage excess sludge in downflow stationary fixed film digesters. AB - The effects of mechanical disintegration on anaerobic digestibility of sewage excess sludge in downflow stationary fixed film (DSFF) digesters were investigated on laboratory scale. Mechanical pretreatment using a high pressure homogenizer led to significantly enhanced concentrations of soluble proteins and carbohydrates in the feed sludge. Using DSFF digesters with two different tubular plastic media as support material it was shown that a stable digestion process could be achieved at hydraulic retention times (HRT) down to 5 days. Compared to conventional digesters at 10 d and 15 d HRT respectively, the degradation of volatile solids was enhanced up to 25%, also resulting in a higher specific biogas production. Further investigations on degradation of soluble proteins and carbohydrates showed that a slowly degradable fraction of carbohydrates was released via disintegration. Using the distribution of chain length and the concentrations of volatile fatty acids as process parameters, the dependability on the HRT and the degree of disintegration (the release of soluble COD) predominated the effects of specific surface area of the support media. PMID- 11381990 TI - Thermal hydrolysis (TDH) as a pretreatment method for the digestion of organic waste. AB - The recycling concept under consideration is based on the process of Thermal Hydrolysis (TDH) followed by an anaerobic digestion. By increasing pressure and temperature the organic part of the waste is split up in a first step into short chain fragments that are biologically well suited for microorganisms. The following fermentation runs much faster and more complete than in conventional digestion processes and the biogas yield is increased. Left is just a small amount of a solid residue that can be easily dewatered and utilized as surrogate fuel for incineration or as compost additive. The thermal hydrolysis process allows a complete energy recovery from organic waste. During the total procedure more energy sources are produced than are needed for running the plant. The procedure is especially suited for wet organic waste and biosolids that are difficult to compost, such as food scraps, biological waste from compact residential areas and sewage sludge. As a complete disinfection is granted due to the process temperatures the procedure is also suited for carcasses. PMID- 11381991 TI - Methods for increasing the biogas potential from the recalcitrant organic matter contained in manure. AB - The biogas potential of manure could be significantly increased by treatment of the recalcitrant organic matter (biofibers) contained in the manure. Several treatment methods were tested. Mechanical maceration resulted in an average increase of the biogas potential of approximately 17% as shown by the continuous stirred reactor experiment. In general the smaller the fibers the higher the biogas potential was. The best results showed an approximately 20% increase of the biogas potential with fibers smaller than 0.35 mm as measured by batch experiments. The increase was approximately 16% with fibers of size 2 mm. Chemical treatment of the fibers with bases such as NaOH, NH4OH or a combination of bases also resulted in an increased methane potential. However, combination of maceration and chemical treatment did not result in a further increase of the methane potential. There was not any significant difference of the biogas potential from fibers in the range 5-20 mm. Treatment of the fibers with hemicellulolytic or cellulolytic enzymes did not result in any significant increase of the methane potential. However, biological treatment of the fibers of the manure with the hemicellulose degrading bacterium B4 resulted in a significant increase of the biogas potential of manure. An increase of approximately 30% in methane potential was achieved compared to controls. PMID- 11381992 TI - Full scale co-digestion of organic waste. AB - Operational results of a co-digestion facility were assessed over a period of 18 months. The organic fraction of municipal solid waste (OFMSW) contains a considerable amount of contaminants and grit (up to 6% w/w). A BTA-Pulper efficiently treated the different waste streams and converted a high amount of volatile solids (VS) into the digester feedstock. The seasonal fluctuations of the waste composition significantly influenced the biogas production. The impact of this seasonally variant degradability of VS had to be considered by evaluating the operation results. The waste streams investigated did not show any negative impact on digester performance. The hydraulic retention time (HRT) in the digester considerably affected the VS-reduction. Despite a considerable decrease of VS-degradation a reduction of HRT from 14 to 8 days slightly improves the gas production rate (GPR). An activated sludge system efficiently reduced the pollution of the effluent. The nutrient content of the anaerobic compost was favourable and the content of pollutants was low. The facility produced surplus electrical power up to 290 MJ/t. An overall energy balance shows that the facility substitutes primary energy. PMID- 11381993 TI - Anaerobic digestion technologies for closing the domestic water, carbon and nutrient cycles. AB - Sustainable wastewater treatment requires that household wastewater is collected and treated separately from industrial wastewater and rainwater run-offs. This separate treatment is, however, still inadequate, as more than 70% of the nutrients and much of the chemical oxygen demand (COD) and potential pathogens of a domestic sewage system are confined to the few litres of black water (faeces, urine and toilet water). Whilst grey water can easily be filter treated and re used for secondary household purposes, black water requires more intensive treatment due to its high COD and microbial (pathogens) content. Recently developed vacuum/dry toilets produce a nutrient rich semi-solid waste stream, which, with proper treatment, offers the possibility of nutrient, carbon, water and energy recovery. This study investigates the terrestrial applicability of Life Support System (LSS) concepts as a framework for future domestic waste management. The possibilities of treating black water together with other types of human-generated solid waste (biowastes/mixed wastes) in an anaerobic reactor system at thermophilic conditions, as well as some post treatment alternatives for product recovery and re-use, are considered. Energy can partially be recovered in the form of biogas produced during anaerobic digestion. The system is investigated in the form of theoretical mass balances, together with an assessment of the current feasibility of this technology and other post-treatment alternatives. PMID- 11381994 TI - Co-digestion of organic solid waste and sludge from sewage treatment. AB - Solid organic wastes were codigested together with sludge of a sewage treatment plant (STP). In the practical part of the study, a plant to pretreat the organic solid wastes provided by local super markets was constructed at the STP of Frutigen, Switzerland. Up to more than 1 cubic metre of wastes was added to the fermenter of the STP every day. Data collected during 14 months of practical works, showed that for raw fruit and vegetable wastes a two step pretreatment is necessary: First the wastes were chopped and afterwards reduced to a size of 1-2 millimetres, in order to get a homogeneous suspension together with the primary sludge. The vegetable wastes showed excellent digestibility: They seemed to accelerate the digestion process as well as to increase the degree of the anaerobic degradation of the sludge. The energy demand for both, pretreatment and digestion, was 85 kWh/ton of fresh wastes. 20% of the energy was used for the hygienization, a step which does not seem to be necessary for this kind of waste in most of the cases, however. After using the gas for energy conversion, a net yield of 65 kWh/ton of electricity and 166 kWh/ton of heat was measured. Treating cooked kitchen wastes, the net energy production will be higher, because in this case a one step pretreatment will be sufficient. The pretreatment and treatment costs for codigestion on STP's were calculated to be in the range of 55 US$/ton treating half a ton per day and 39 US$/ton treating one ton, respectively. A theoretical feasibility study showed that in Switzerland there is a short term potential on STP's for the codigestion of about 120,000 tons of biogenic wastes per year without big investments. Economic studies about codigestion on agricultural biogas plants showed that the codigestion is a must at the current energy prices, which are far too low for agricultural AD without an additional income by treating solid wastes for third parties. PMID- 11381995 TI - Characterization and anaerobic treatment of the sanitary landfill leachate in Istanbul. AB - In this study, characterization and anaerobic treatability of leachate from Komurcuoda Sanitary Landfill located on the Asian part of Istanbul were investigated. Time based fluctuations in characteristics of leachate were monitored for an 8 month period. Samples were taken from a 200 m3 holding tank located at the lowest elevation of the landfill. COD concentrations have ranged between 18,800 and 47,800 mg/l while BOD5 between 6820 and 38,500 mg/L. COD and BOD5 values were higher in summer and lower in winter due to dilution by precipitation. On the other hand, it was quite interesting that such a dilution effect was not observed for ammonia. The highest ammonia concentration, 2690 mg/L was in November 1998. BOD5/COD ratio was larger than 0.7 for most samples indicating high biodegradability, and acidic phase of decomposition in the landfill. For anaerobic treatability, three different reactors, namely an upflow anaerobic sludge bed reactor, an anaerobic upflow filter and a hybrid bed reactor, were used. The anaerobic reactors were operated for more than 230 days and were continuing operation when this paper was prepared. Organic loading was increased gradually from 1.3 kg COD/m3.day to 8.2 kg COD/m3.day while hydraulic retention time was reduced from 2.4 days to 2.0 days. All the reactors showed similar performances against organic loadings with efficiencies between 80% and 90%. However the reactors have experienced high ammonia concentrations several times throughout the experimental period, and showed different inhibition levels. Anaerobic filter was the least affected reactor while UASB was the most. Hybrid bed reactor has exhibited a similar performance to anaerobic filter although not to the same degree. PMID- 11381996 TI - Kinetics of accelerated solid-state fermentation of organic-rich municipal solid waste. AB - Biotransformation of landfill solid wastes is a slow process requiring decades for completion. Accelerated anaerobic fermentation in modulated landfill environments may alleviate or eliminate pollution of land, water and air. This research was undertaken to demonstrate the application of biphasic fermentation to a simulated laboratory-scale landfill to effect rapid biomethanation of biodegradable solids. The biphasic process consisted of solid-state, acidogenic fermentation of the organic fraction of MSW followed by biomethanation of acidic hydrolysates in a separate methane fermenter. Solid-state fermentation of the MSW with effluent recirculation resulted in rapid hydrolysis, acidification and denitrification, with soluble COD and VFA concentrations accumulating to inhibitory levels of 60,000 mg/l and 13,000 mg/l, respectively, at a pH of 4.5. The landfill gas methane concentration reached a maximum of 55 mol.%. By comparison, the methanogenic reactor produced high methane-content (70-85 mol.%) gases. The biphasic process effected carbohydrate, lipid, and protein conversion efficiencies of 90%, 49%, and 37%, respectively. Development of a Monod-type product-formation model was undertaken to predict methane formation and to determine kinetic parameters for the methanogenic processes in the simulated landfill and separate methane reactors. A first-order solids hydrolysis rate constant of 0.017 day-1 was evaluated to show that landfill solids hydrolysis was slower than the inhibited methanogenesis rate. PMID- 11381997 TI - Anaerobic treatment of landfill leachate by sulfate reduction. AB - The present study was conducted to investigate the effectiveness of the sulphate reduction pathway in the anaerobic treatment of landfill leachate. The effects of several COD/SO4 ratios (keeping COD constant) and loadings on anaerobic filter performance were studied and compared with the results from anaerobic filters which followed the methanogenic pathway. Results indicated that the treatability of leachate by sulphate reducing bacteria (SRB) was dependent upon the leachate strength. With high strength leachate (COD = 15,000 mg/L) from the Keele Valley Landfill, it was found that at lower COD/SO4 ratios (< or = 1.6) toxic conditions developed in the system that were more inhibitory to the SRB than to the methane producing bacteria (MPB). As the COD/SO4 ratio increased, methanogenesis predominated. No predominance of SRB occurred at any COD/SO4 ratio with high strength leachate. The highest COD removal achieved was about 70% of which 20% was accomplished by the SRB at a COD/SO4 ratio of 1.6 and an organic loading rate (OLR) of 4 kg COD/m3.d. With low strength leachate (COD = 1500-3300 mg/L) from the Brock West Landfill, and a COD/SO4 ratio < or = 1, SRB became predominant. In these anaerobic filters in which SRB were predominant, the SRB reduced the COD as well as the MPB could. Sulphide inhibition did not take place at any loading in units treating low strength leachate. Consequently, both SRB and MPB should function at COD/SO4 ratios between 1 and 3. About 60% COD removal was achieved at a loading of 2.8 kg COD/m3.d and a COD/SO4 ratio of 1.0. However at a loading of 6 kg COD/m3.d only 27% COD removal was achieved, all of it through the sulphate reduction pathway. These OLR values are comparable to those applied in systems where methanogenesis was dominant. It was also observed that once the methanogens were established in the units, it was not possible to displace them completely. However, where methanogenesis had not been previously established, it was found that sulphate-reduction could be the sole pathway for COD removal. From this study, it can be concluded that there is no advantage to the sulphate-reduction pathway in the anaerobic treatment of landfill leachate. The other options for increasing the loadings, i.e. the use of high surface/volume filter media (to achieve higher biomass concentrations) or high rate systems are likely to be more successful. PMID- 11381998 TI - Microbially mediated attenuation potential of landfill bioreactor systems. AB - The origin and fate of landfill leachate and gas constituents generated during the sequential phases of solid waste transformation and stabilization are emphasized within the perspective of the in situ processes of microbially mediated attenuation. The fundamental biochemical and physicochemical reaction mechanisms are presented in terms of their spatial and temporal dimensions and their significance for transformation of both nonhazardous and hazardous waste constituents. Supporting information from laboratory, pilot-scale and full-scale applications is used as a basis for interpretive analysis and for providing operational guidance and promoting future developments. The diversity, domains, and functional interdependence of the acidogenic, methanogenic, sulfate and nitrate reducing, nitrifying and denitrifying, and methanotrophic consortia are addressed in order to reveal opportunities for landfill process modifications and associated operational optimization. Controlled attenuation, linked with operational and regulatory realities, are used to suggest innovative landfill configurations involving prospective compartmentalization and integrated waste loading, dedicated treatment zones for in situ transformation of waste and leachate constituents with associated gas capture, control and utilization. Monitoring requirements are emphasized to provide guidance and feedback for operational control and environmental compliance. Finally, technology needs for establishing a more unified approach to the development and management of bioreactor landfills are presented. PMID- 11381999 TI - Biological hydrogen potential of materials characteristic of the organic fraction of municipal solid wastes. AB - The purpose of this study is to investigate the biological hydrogen production potential of individual organic fraction of municipal solid wastes (OFMSW) by batch experiments. Seven varieties of typical organic solid wastes including rice, cabbage, carrot, egg, lean meat, fat and chicken skin were selected to estimate the hydrogen production potential. Among the OFMSW, carbohydrate produced the most hydrogen through biological hydrogen fermentation compared with proteins or lipids. Subsequently, the biological hydrogen production potentials of some individual carbohydrate were measured: cabbage, 26.3-61.7 mL/g-VS; carrot, 44.9-70.7 mL/g-VS; and rice, 19.3-96.0 mL/g-VS. The hydrogen percentages of the total biogas produced from cabbage, carrot and rice were 33.9-55.1%, 27.7 46.8% and 44.0-45.6%, respectively. PMID- 11382000 TI - Effect of substrate-seed mixing and leachate recirculation on solid state digestion of biowaste. AB - Lab-scale experiments were performed and a mechanistic model was developed to simulate the solid state digestion of biowaste in a batch reactor. Both experiments and model showed that the substrate-seed mixing degree and leachate recirculation rate have a strong effect on the reactor performance. This is due to mass transport limitations of volatile fatty acids (VFA) within the biowaste seed bed. In that case two regions are developed in the digester, so-called acidogenic and methanogenic pockets. Limitations in mass transport will prevent irreversible acidification during start-up of the reactor because whereas high VFA concentration is met in the fresh waste pockets, the VFA concentration in the methanogenic pockets will remain low. However, accumulation of VFA in the acidogenic pockets will reduce the hydrolysis rate of biowaste due to inhibition by VFA. Moreover, experiments and simulations showed that the reactor performance can be improved by varying the leachate recirculation rate or applying sequential batch operation. PMID- 11382001 TI - Ecological, energetic and economic comparison of anaerobic digestion with different competing technologies to treat biogenic wastes. AB - In order to get more detailed information for better decision making in future biogenic waste treatment, different processes to treat biogenic wastes in plants with a treating capacity of 10,000 tons of organic household wastes per year were compared. The comparison included life cycle assessments as well as economic considerations for different treating methods. Measurements on compost plants showed that methane emissions are higher than estimated so far. With the tools ECOINDICATOR and UBP anaerobic digestion shows to be advantageous as compared to composting, incineration or combination of digestion and composting, mainly because of a better energy balance. In fully enclosed, professional treatment plants, the specific biotechnological treatment costs are in the range of about 150.-sFr/ton for aerobic, anaerobic and combined technologies. It can be concluded, that anaerobic processes will become considerably more important in the future mainly for ecological reasons. PMID- 11382002 TI - Emission of greenhouse gases from anaerobic digestion processes: comparison with other municipal solid waste treatments. AB - This contribution analyzes the anaerobic digestion process and compares GHG emissions estimated for four different management processes for MSW (Municipal Solid Waste): biogasification, landfilling, composting and incineration. The comparison has been undertaken by considering in the estimation of the emissions the full cycle of MSW treatment, and not only the emissions derived from the fraction of MSW treated by each particular system. For instance, the fraction of MSW not submitted to biological treatment has to be incinerated or deposited in a landfill. The corresponding emissions of these processes have to be considered in the calculations of the final emissions. PMID- 11382003 TI - Anaerobic digestion of solid waste: state-of-the-art. AB - In order to make a correct assessment of the state-of-the-art of the technology, a study was made on the development of digestion capacity for solid waste in Europe. The study was limited to plants in operation or under construction that were treating at least 10% organic solid waste coming from market waste or municipal solid waste. A total treatment capacity for solid waste organics, excluding the tonnage used for sewage sludge and manures, evolved from 122,000 ton per year in 1990 to 1,037,000 ton available or under construction by the year 2000 in 53 plants across Europe, an increase by 750%. Both mesophilic and thermophilic technologies have been proven, with about 62% of capacity being operated at mesophilic temperatures. Wet and dry digestion are almost evenly split, while a clear choice was made for one-phase systems instead of two-phase systems, which represent only 10.6% of capacity. The capacity provided by co digestion systems is limited, while there is a rising interest in digestion of mixed household waste. The reliable performance has been demonstrated for all types of anaerobic digestion systems. On the basis of the Dranco technology, a single-phase thermophilic dry digestion process, performances were reached similar to high-rate wastewater digestion. An annual average loading rate of 18.5 kg COD/m3.day, resulting in a biogas production of 9.2 m3/m3 reactor.day was obtained at a full-scale plant. The plant operated at a retention time of 15.3 days. Feedstocks range from clean organic wastes (31% dry matter) to heavily polluted grey waste organics (57% dry matter). Average dry matter concentrations of the digested residue of 41% were obtained. PMID- 11382004 TI - Determinant impact of waste collection and composition on anaerobic digestion performance: industrial results. AB - The performance of the anaerobic digestion process depends deeply on the quality of the waste to be treated. This has been already demonstrated at the lab-scale. The objective of this study is to confirm this result at the industrial scale, with very long representative period and with the same process, the Valorga process. According to the waste quality and the collection type and even with the same conditions of fermentation, the biogas yield can vary by a factor of 1.5 when it is expressed (under normal conditions of pressure and temperature) in m3 biogas/t fresh waste, and by a factor of 2 when it is expressed in m3 CH4/t volatile solids. So, the biogas performance does not characterise a process since it is deeply governed by waste composition. This biogas productivity becomes a pertinent parameter only with consistent and relevant hypothesis and/or analytical results on the waste composition which depends on the collection procedure, the site characteristics and the season. PMID- 11382005 TI - Full scale experience with the BIOCEL process. AB - The BIOCEL process is a mesophilic dry anaerobic batch digestion system for solid organic wastes. In the BIOCEL process organic solid wastes, such as source separated organic fraction of MSW (biowaste) is converted into enriched compost and biogas. In the process net energy production is achieved by converting the biogas to heat and power with a heat-electric power production unit. In September 1997 the first full scale plant is started-up in Lelystad, The Netherlands. This plant is processing 50,000 tons of biowaste (organic fraction of MSW from source separation) per year. The plant has a net energy production and therefore contributes to prevention of CO2 emissions from fossil fuels. In the BIOCEL system the several compost fractions are produced with a "wet" separation process. During the wet separation sand and contaminants are removed. An important aspect of compost quality is the absence of several types of pathogens. It appears that anaerobic digestion with the BIOCEL-process results in complete inactivation of several important groups of plant and animal pathogens. The mechanism that causes the inactivation is not yet fully understood, but the relatively high Volatile Fatty Acids concentration during the first two weeks of the digestion process might presumably be the key factor. PMID- 11382006 TI - Establishing procedures for design, operation and maintenance of sewage sludge anaerobic treatment plants. AB - In 1993 the Irish Government commissioned a strategy study to address the difficulties encountered with sludge management and disposal in this country. Many Local Authorities have complied with the recommendations of this strategy study by incorporating Anaerobic Digestion (AD) of sewage solids in the design of new and retrofitted sewage treatment plants. Irish Local Authorities have little experience in anaerobic technologies compared to the more traditional mechanical and aerobic processes. The United States Environmental Protection Agency (US EPA) developed procedures for evaluating sewage treatment plants. These Comprehensive Performance Evaluations (CPE) are described in such publications as "Retrofitting Publicly Owned Treatment Works (1989)". The emphasis in such documentation was on aerobic secondary treatment processes. The present study developed an evaluation procedure for AD plants similar to the CPEs produced by the US EPA for secondary processes. This procedure was applied to existing and newly commissioned AD plants in Ireland in order to deduce process performance. Based on these studies, recommendation for the design, operation and maintenance of AD plants is presented. All plant operators agreed that frequent and accurate monitoring of pH and temperature was essential for adequate process control. The five existing sites were surveyed. Apart from the usual difficulties at start-up, plant operators expressed satisfaction with the anaerobic process at four of the sites. The fifth plant is currently being upgraded. PMID- 11382007 TI - Two-particle model of anaerobic solid state fermentation. AB - A structured mathematical model of anaerobic solid state fermentation (ASSF) has been developed. Since a stable ASSF requires addition of significant quantities of methanogenic seed sludge and mass-transfer limitation becomes important, the model postulates the existence of two different types of particles inside the fermenting solid mass--so-called "seed" particles with low biodegradability and high methanogenic activity and so-called "waste" particles with high biodegradability and low methanogenic activity. Any particle is assumed to be a completely mixed reactor and mass transfer of solutes between the particles is brought about by diffusion. The model includes multiple-reaction stoichiometry, microbial growth kinetics, material balances, liquid-gas interactions and liquid phase equilibrium chemistry. The theoretical model agrees on the qualitative level with existing experimental studies of ASSF. Hypothetical computer simulations are presented to illustrate the influence of biodegradability and mass transfer intensity on the stability of ASSF. On this basis, possible measures are proposed to prevent accumulation of volatile fatty acids inside the "seed" particles beyond their assimilative methanogenic capacity. PMID- 11382008 TI - Rheology of sludge from double phase anaerobic digestion of organic fraction of municipal solid waste. AB - In this paper experimental results on the anaerobic digestion of sewage sludge and organic fraction of municipal solid waste (OFMSW) by using a double phase process are reported. The long-term experiment has been carried out on a pilot scale plant, performed in different sets of operative conditions, during which granulometric distributions of particles in sludges and rheological properties of sludges were monitored. A significant fluidification of sludge was evidenced in the meso-thermo process, especially taking into account the variation in sludge behaviour from the first to the second phase. In the thermo-thermo process a fluidification higher than that shown in meso-thermo conditions is not observed, this suggesting that better results in terms of sludge conditioning can be obtained in a long time spent in thermophilic anaerobic digestion. Total volatile solids (TVS) and total fixed solids (TFS) become the most important parameters when mathematical modelling is applied to these processes. In the acidogenic phase, hydraulic retention time (HRT) and temperature are used to determine rigidity coefficient (RC), while only temperature is needed for yield stress (YC). Organic loading rate (OLR) and specific gas production (SGP) exert an important role in methanogenic phase description. PMID- 11382009 TI - Mathematical modeling of the hydrolysis of anaerobic processes. AB - In the recently published dynamic simulation model for mesophilic digestion of sewage sludge the hydrolysis constant refers to the total dry solid content without regarding their composition. To apply these models to the digestion of municipal solid waste, the hydrolysis constants of the various fractions must be considered. The major constituents of organic waste were identified as carbohydrates, proteins and lipids. For these three constituents the hydrolysis constants for thermophilic digestion were determined. The implementation of these constants into the existing dynamic models allowed a reasonable characterization of the digestion of municipal organic waste. PMID- 11382010 TI - Effect of particle size and sodium ion concentration on anaerobic thermophilic food waste digestion. AB - Serial basic tests were conducted for the determination of fundamental kinetics and for the actual application of kinetic parameters to food waste digestion with precise measurement of methane production. The effects of food particle size and sodium ion concentration on the anaerobic thermophilic food waste digestion process were investigated. Results of serial tests for the determination of fundamental kinetic coefficients showed the value of k (maximum substrate utilization rate coefficient) and KS (half-saturation coefficient) as 0.24 hr-1 and 700 mg/L, respectively, for non-inhibiting organic loading range. The substrate inhibition coefficient (Ki) was 1000 mg/L for inhibiting organic loading range. No inhibition effect was shown until 5 g/L of sodium ion concentration was applied to the serum bottle reactor. However, the volume of methane gas was decreased gradually to the concentrations of more than 5 g/L of sodium ion applied. All sizes of food waste particle had the same constants (A: 0.45) but the maximum substrate utilization rate constant (kHA) was inversely proportional to particle size. As average particle size increased from 1.02 mm to 2.14 mm, kHA decreased from 0.0033 hr-1 to 0.0015 hr-1. Result reveals that particle size is one of the most important factors in anaerobic food waste digestion. PMID- 11382011 TI - Performance of thermophilic semi-dry anaerobic digestion process changing the feed biodegradability. AB - The study concerns the application of the semi-dry single phase thermophilic anaerobic digestion process to the organic fraction of municipal solid waste. The experiments were carried out using 3 m3 and 1 m3 CSTR pilot scale reactors. The process performance in terms of biogas yields, digester stability and kinetic aspects was studied, considering a progressive increase in the feed biodegradability, in order to evaluate the process behaviour changing from an undifferentiated collection of waste to a separate collection. This was carried out using blends of two different kinds of substrates: mechanically sorted organic fraction of municipal solid wastes (MS-OFMSW) and source sorted organic fraction of municipal solid wastes (SS-OFMSW). The study shows that OLR up to 6 kg TVS/m3d can be applicable for the medium selected fraction (TVS/TS < or = 0.7), while for the MS-OFMSW alone this limit can be doubled. The results obtained with SS-OFMSW alone suggest the use of the double phase process to give more stable conditions. PMID- 11382012 TI - Development of a methanogenic process to degrade exhaustively the organic fraction of municipal "grey waste" under thermophilic and hyperthermophilic conditions. AB - Different laboratory-scale, continuously driven reactor concepts (up to 3 reactors in series, max. 70 degrees C) for anaerobic digestion of the organic fraction of municipal grey waste were investigated. Over a period of 2 1/2 years several setups of reactors being daily fed and held in steady state balance were investigated. The perferred variant was a 2-stage setup with a HRT of 4.3 d for the 1st and 14.2 d for the 2nd reactor. Removal efficiencies of VS obtained by comparing the organic loading rate (OLR, g VS/l/d) of the effluent with the OLR of the feed could reach 80%. Removal efficiencies determined indirectly by the combined biogas yield of the 1st and 2nd reactor stage revealed even up to 91.5% of the theoretical possible yield of 807 l/kg VS. The produced gas had a methane content of 60-65%. A completely distinct hydrolysis stage with a gas production of only 1.6-5.5% of the theoretical yield could be reached by hyperthermophilic conditions (60-70 degrees C) or by a HRT of 1.25 d. It also demonstrated that a stable methanogenesis was not possible at temperatures of 60-70 degrees C. Kinetic analyses of the 2nd reactor stage revealed that the degradation of VS fell from 80 to 40% with raising organic loading rate (OLR) from 3 to 11 g VS/l/d. In contrast to this the VS-removal of the first hydrolysis reactor stage increased linearily from 5 to 20% at raising OLR's from 12 to 26 g VS/l/d. The same kinetics with linear increase exhibited the specific cellulose degradation with conversion rates of 0.1-3 x 109 g cellulose/single bacterium (10(-12) g)/d. This was an indication for the cellulose degradation as a rate limiting step. Both reactor stages combined allowed an optimal VS removal efficiency at OLR of 10 g VS/l/d. Analysis of bacterial populations of 28 reactors were referred either to eubacteria utilizing different sugars or cellulose or acetate or H2-CO2 or archaea (plus antibiotics) with acetate or H2-CO2 as substrate. H2-CO2 utilizers with numbers of 10(8)-10(10)/g TS dominated obviously the acetotrophic methanogens by the factor 10-10,000. This explained the observed short HRTs being possible. PMID- 11382013 TI - Microbial ecology of the leach bed anaerobic digestion of unsorted municipal solid waste. AB - The microbial ecology of the sequential, leach-bed, mesophilic anaerobic digestion of unsorted, coarse municipal solid waste (MSW) was examined over 80 days. The methane yield was approximately 75% of the ultimate biochemical methane potential (BMP) of the waste loaded into the digesters. The operational strategy involved a sequence of two digesters containing fresh and anaerobically stabilised MSW respectively. Cell wall phospholipid fatty acid (PLFA) and ether lipid (PLEL) analysis was used to monitor changes in microbial biomass. Both Bacterial and Archaeal biomass were heavily influenced by pH during the two-week start up period. Archaeal biomass peaked just before the methane production rate reached a maximum whereas Bacterial biomass peaked at a later stage. Changes in the phylogenetic diversity of the population were monitored by denaturing gradient gel electrophoresis (DGGE). An analysis of the changes in DGGE banding patterns suggested that rapid start-up of a new reactor was effected by inoculation as well as the provision of buffering capacity from the mature reactor leachate. PMID- 11382014 TI - Bench-scale anaerobic bioconversion of newsprint and office paper. AB - Anaerobic bioconversion of newsprint and waste office paper was performed in bench-scale reactors with three inocula sources: landfill, rumen, and anaerobic digester. Office paper bioconversion was nearly complete within 20 days but continued for about 165 days with methane yield efficiencies ranging from 71-85% of potential chemical oxygen demand (COD) conversion. Average newsprint methane conversion efficiencies ranged from 32-41% of total COD under strictly anaerobic conditions for 300 days. Mass balance calculations revealed that more than 80% of newsprint cellulose was biodegraded. The apparent limiting factor for anaerobic bioconversion of newsprint was the physical association between lignin and cellulose. After proper acclimation, the three inocula tested equally well for methane production under strictly anaerobic conditions. Testing of ground, shredded strips, and whole paper pieces showed no effects of feedstock size on bioconversion rate or extent. Alkali pretreatment with NaOH concentration up to 10% significantly improved newsprint biodegradability. Treatment for longer duration or at elevated temperatures increased the solubilization of lignin, but did not improve bioconversion of newsprint to methane. Neutralizing treated samples with carbon dioxide gave higher methane yields compared to sulfuric or hydrochloric acids, suggesting that digester neutralization could be combined with biogas scrubbing. PMID- 11382015 TI - Effect of bioaugmentation on microbial transport, water infiltration, moisture loss, and surface hardness in pristine and contaminated soils. AB - Three different soils, a clay, a pristine sandy loam and a PCB-contaminated sandy loam, were bioaugmented to determine the influence of clay content and contaminants on the transport of bacteria in unsaturated soils, using surface irrigation water as a transport medium. The results indicate that the transport of the implanted bacteria was influenced negatively more by the presence of PCBs than by the clay content of the soil. Transport was directly related to the frequency of irrigation and length of the intervals between irrigation periods, making these variables important factors to consider when applying bioaugmentation via downward percolating water. Other parameters measured after bacterial bioaugmentation were water infiltration, moisture loss, and surface hardness of these soils. Surface water infiltration was affected more by the soil clay content than by the hydrophobic contaminant. Infiltration was significantly but differently influenced by bioaugmentation, positively in clay, negatively in sandy loam, and negatively (to a lesser extent) in the PCB-contaminated sandy loam soils. Moisture loss was slower in the bioaugmented soil than in the control soils, with this difference being most pronounced in the PCB soil. High moisture loss in the bioaugmented clay soil rendered it the hardest soil for surface penetration. PMID- 11382016 TI - Interaction of nitric oxide with the surface of stabilized calcium sulfate. AB - This work explores surface interactions between stabilized gypsum and nitric oxide. Gypsum is a common air-borne mineral particulate that has a potential two fold relationship to the air pollution problem: as a particulate pollutant and as a catalyst or adsorbent for pollutant gases. Nitric Oxide is found in stack gases and automotive exhausts. Isotherms of nitric oxide adsorbed on stabilized gypsum were studied at 24 degrees C, 0 degree C, and -78 degrees C. Coverages were related to a monolayer based upon a surface area of 17.0 m2/g as determined from a nitrogen adsorption isotherm and the B.E.T. method. Multilayer coverages containing both reversible and irreversible adsorption were observed for nitric oxide adsorbed on stabilized hydrated calcium sulfate. An irreversible coverage of 41% at 24 degrees C and 61% at 0 degree C of the nitrogen monolayer was observed for nitric oxide adsorbed on hydration stabilized gypsum. The heat of adsorption at zero coverage was found to be 80.4 kJ/mol for nitric oxide on stabilized hydrated calcium sulfate for the irreversible adsorption and 7.5 kJ/mol for the reversible adsorption. PMID- 11382017 TI - Statistical evaluation of PCDD/F emission data during solid waste combustion by fuzzy clustering techniques. AB - An advanced statistical analysis technique using the fuzzy clustering method was employed in this work, for the evaluation of PCDD/F emissions during solid waste combustion. In addition, this technique was applied for the assessment of the effect of an inhibitor (urea) on the toxic compound releases and on the various isomer distributions. Municipal solid wastes were combusted in a lab-scale reactor and the toxic gas emissions were measured at the unit outlet. Combustion tests of urea-fuel mixtures were classified in the same group, indicating that urea affected the formation mechanisms of toxic gases. Combustion tests of single fuel were not included in the same group. Furthermore, urea ability to modify the gas emissions pathways was not affected by the method of its addition to the fuel. PMID- 11382018 TI - Catalytic degradation of high density polyethylene using zeolites. AB - Plastic wastes, which cause a serious environmental problem in urban areas, can serve as sources of energy. Catalytic treatment of High Density Polyethylene (HDPE) has shown that the degradation of HDPE resulted in the production of a stream of gaseous hydrocarbons varied in the range C1-C8. The degradation was carried out using diluted forms of zeolites ZSM-5, USY and Mordenite (MORD) using a fluidized bed reactor (FBR). Effect of coke formation on the activity of the catalysts was screened by thermogravimetric (TGA). ZSM-5 showed a significant resistance to deactivation because of the nature of its small pore size compared with USY and MORD. PMID- 11382019 TI - The comparative selection between spiral wound and disc tube membranes to treat steel industry wastewater. AB - To select the most suitable membrane process to treat the final effluent discharged from the S wastewater treatment facility at P Steel Works, the effluent was passed through two types of membrane (spiral wound and disc tube) in a pilot-scale study. The permeate flux and regeneration of each membrane were comparatively observed and the removal efficiency of contaminants determined. The experimental results from employing the disc tube membrane revealed that the fouling was comparably limited by operating the plant at 34 bar recovery and was highly sustained at permeate flux of 75% which equals to 32.9 l/hr.m2. In contrast, the fouling was significantly greater when applying the spiral would membrane with the recovery correspondingly diminished at 72%. The disc-tube membrane was chosen to treat the effluent to be recycled for process water as the water quality satisfied the industry water standards. PMID- 11382020 TI - Enhancing the dewaterability and amenability of sludge for subsequent stabilization processes by using organic waste solids as conditioners. AB - Dual-material conditioning of sludge was evaluated using a chemical conditioner (either alum or ferric chloride), followed by a physical conditioner (either wood chips or wheat dregs). The combined effects of the conditioners on the dewaterability of the sludge with vacuum filtration and the amenability of the resulting sludge cake for subsequent stabilization processes were investigated. Results showed that the specific resistance of the conditioned sludge was effectively reduced when the dose of wood chips or wheat dregs was increased in each dual-material combination. From the results of analysis of solids in the cake, the filtered sludge cake with either wood chips or wheat dregs conditioning in higher dosage resulted in lower moisture and higher volatile combustible solids content. Accordingly, sludge conditioning with these two materials substantially increased the heating value of the dewatered sludge. In addition, wood chips are the most widely used bulking agent, and wheat dregs could be suitable to act as amendments in sludge composting. These results indicate that these two organic waste solids could replace a significant portion of the chemicals now used for sludge conditioning to improve the dewaterability of sludge and the amenability of the resulting sludge cake for incineration and composting processes. PMID- 11382021 TI - Kinetics of photocatalytic degradation of reactive yellow 17 dye in aqueous solution using UV irradiation. AB - In this paper photoinduced degradation of reactive yellow 17 (RY 17) dye has been studied employing TiO2 in the form of slurry in a batch reactor. UV lamps were used as the source for irradiation. The disappearance of the dye in the solution follows approximately pseudo-first order kinetics. The apparent rate constant decreases with increase in initial concentration of the dye. The addition of oxygen, hydrogen peroxide and persulphate ion influences the degradation rate of the dye. The formation of CO2, SO4(2-), NO3- and NH4+ ions have been identified in the mineralisation process. The decolourisation and mineralisation were followed by UV-Visible spectrophotometer and high performance liquid chromatography respectively. The results reveal that photocatalytic degradation may be a useful technique in the removal of reactive dyes in wastewater from textile industries. PMID- 11382022 TI - Source contributions to ambient concentrations of CO and NOX in the urban area of Beijing. AB - An emission inventory based on GIS technology was developed in this study, and used to estimate the spatial distribution of the stationary and mobile sources in the urban area of Beijing. The stationary sources were divided into various types of finer classes depending on different usage, stack height and emission factors. The mobile sources were treated as both line sources and area sources. It was shown that total anthropogenic CO and NOX emissions had reached 1.4 million and 233 thousand tons, respectively, and vehicle sources emissions of CO and NOX accounted for 76.8% and 40.2%, respectively, of the total emissions in 1995. ISCST3, a Gaussian dispersion air quality model, was modified at low wind speed condition, and then used to facilitate the study of source contributions to ambient concentrations of CO and NOX in Beijing. The simulation results showed that emissions from the vehicle sources had contributed 76.5% and 68.4% of the total CO and NOX concentrations in urban atmosphere of Beijing in 1995, and were even higher in the city core (86.3% of the CO and 72.0% of the NOX). Therefore, strategies for CO and NOX pollution control will of necessity need to focus on the vehicle sources. PMID- 11382023 TI - The effect of surfactants on the deliquescence of sodium chloride. AB - This study investigated the deliquescence of sodium chloride aerosol with surfactants at a retention time of 4.24 sec. Two surfactants used in this study, glutaric acid and pyruvic acid, are found in atmospheric aerosol and have a high hydrophile-lipophile balance (HLB) value. The experimental system consisted of a relative humidity conditioner, a tandem differential mobility analyzer (TDMA) and a scanning mobility particle sizing (SMPS) system. Results obtained from the observation of TDMA presented the deliquescence point of sodium chloride aerosol at 75% RH. In addition, the growth size of sodium chloride aerosol was observed to be 79.47 nm and the growth ratio was 77.94%, when the initial size of aerosol was 101.82 nm. Surfactants were internally mixed with sodium chloride at six different weight fractions, i.e. 2.5, 5, 10, 20, 40, and 60% by weight of surfactants in dry aerosol. Both surfactants apparently decreased the deliquescence point of sodium chloride, in which the lowest deliquescence point appeared at about 71% RH when the weight fraction of surfactants is 60% by weight of surfactants in dry aerosol. Moreover, the smallest size of sodium chloride aerosol with surfactants was 142.7 nm at 60% by weight of pyruvic acid in dry aerosol. Our results further demonstrate that the deliquescence point and size of sodium chloride aerosol with surfactants are related to the weight fraction of surfactants. PMID- 11382024 TI - Kinetic analysis of inhibitory substrate degradation in bioaugmented activated sludge process. AB - A steady-state model coupling Haldane kinetics and mass conservative principles was developed herein to depict the degradation of a single inhibitory substrate in a bioaugmented activated sludge reactor. The substrate toxicity impact on the bioaugmentation was evaluated for three levels of inhibition coefficient Ki: 0.1, 10, and 1000 mg/L. Under high toxicity conditions of Ki = 0.1 mg/L, the input biomass markedly enhanced the substrate removal. However, at less toxicity of Ki > 10 mg/L, the input biomass was ineffective. The sensitivity study confirmed the parameters of k, Ks, and bd has no connection with Ki, but depends on the amounts of degradative bacteria inside the reactor. Alternatively, parameter Y is important only when Ki is greater than 10 mg/L and the bacteria significantly utilize the substrate. In addition, a novel indicator of marginal solids retention time examines the economic advantages of adding biomass as compared to extending the solids retention time. PMID- 11382025 TI - [Influence of environmental factors on CH4 emission from Reed Wetland]. AB - Determination of CH4 emission from reed wetland in Liaohe delta, China using closed chamber(enclosure area 0.64 m2) technique showed that average emission rate of CH4 was 520 micrograms.m-2.h-1 with significant seasonal patterns. When soil redox potential reached -110 mV, CH4 occured and increased with its decrease. CH4 emission was also influenced by water management and reduced with deep water cover. During the period of measurement, there was positive relation between temperature and CH4 flux(R2 = 0.196, n = 21, P < 0.05). In addition, the experimental result also showed that the methanogenic activites in 0-5 cm soil layer were higher and decreased obviously with soil deepening. PMID- 11382026 TI - [Analysis on effect of inspection and maintenance (I/M) program for vehicle emission reduction]. AB - An Inspection and maintenance(I/M) programme aims to ensure that motor vehicle emission control systems are functioning properly throughout the two stage idle test. Based on three factors including data from inspection, standard for inspection and failure rate, the effectiveness of the current I/M program was studied through the cumulative distributions. The disadvantages of the cutpoints, institution settings, management, and supervision was also analyzed. The fundamental law of emission limits was proposed. PMID- 11382027 TI - [Release character of phosphorus from the sediments of east Lake, Wuhan]. AB - Effects of environmental factors on the release of phosphorus were studied at laboratory simulated test. The relationship between the different forms of phosphorus in sediments and the quantity of releasing phosphorus was studied at the same time. Results showed that the rates of phosphorus release from the sediment increased with rising the temperature of disturbing the overlying water. The content of phosphorus release from the sediment reached the lowest level when pH value was 7.4, high pH or low pH increased the rates of phosphorus release from the sediment. In anaerobic, the release rates of phosphorus was 30 times as much as in aerobic. Research also showed that different chemical forms of phosphorus in sediments were related to the quantity of phosphorus released from the sediments in varying degrees. The dissolved phosphorus and Fe binding phosphorus in sediments played the most important roles (r > 0.90, P < 0.01). PMID- 11382028 TI - [Photooxidation of acridine orange in homogeneous system containing Fe3+ and H2O2 under visible irradiation]. AB - It was found that acridine orange dye can be effectively degraded under visible irradiation (> 470 nm). The influence of ferrous ion, oxalate or surfactant DBS was also studied, the results showed that the addition of ferrous ions or DBS may enhance the degradation rate, on the contrary, the addition of oxalate decreased the degradation rate. PMID- 11382029 TI - [Analysis of toxic organic compounds in MGP-Wastewater and investigation of Wastewater's general characteristics]. AB - A full-scale investigation and analysis of MGP(Manufacture Gas from Petroleum) Wastewater was carried out in this study. It's the first time that this paper publicizes the wastewater's general data, pretreatment method of GC/MS analysis and compositions of toxic organic compounds. The concentrtion of wastewater's CODCg, BOD5, phenolic, N-NH4+ and extractive-organic is: 400 mg.L(-1)-600 mg.L-1, 60 mg.L(-1)-80 mg.L-1, 20 mg.L(-1)-30 mg.L-1, 60 mg.L(-1)-80 mg.L-1 and 170 mg.L( 1)-200 mg.L-1 respectively. There are 79 kinds of aromatic compounds were detected by GC/MS, the total concentration of influent and effluent is 140.79 mg.L-1 and 129.11 mg.L-1 respectively. The data approved that MGP-wastewater is toxic and difficult for bio-treatment. PMID- 11382030 TI - [The sedimentation rate and fractional dimension during flocculation with modified natural polymer and polymeric aluminum chloride]. AB - The sedimentation rate and fractional dimension of the floc was studied during the process of the organic waste water treatment with modified natural polymer (CMS) and polymeric aluminum chloride (PAC). Irregular garrulous form of the floc was observed with microscope and electron-scanning microscope. The results showed that the sedimentation rate was 2.6 mm/min with the modified natural polymer (CMS) and 1.7 mm/min without CMS in condition of keeping the pH value to 6.86 and added quantity of PAC to 80 mg/L. The fractional dimension of the floc Df was calculated to be 1.70 in the best sedimentation rate. PMID- 11382032 TI - [Cumulative impact assessment: problems and practice in China mainland and Hong Kong]. AB - Including past, present and future impacts in environmental impact assessment (EIA) of the proposed action, it is the important property of cumulative impact assessment (CIA) that different from traditional EIA. It is also the main trend of EIA improvement. From the area of regulation demands, documentation practice, spatial and temporal boundary, key contents and mitigation measures, this paper analyses the status of CIA both of China Mainland and Hongkong in the practice of EIA. The author suggested that technical guidelines for cumulative effects assessment should be set up in China Mainland and Hongkong, CIA should be included in project, region and strategy environment assessment and cumulative impacts should be list clearly in EIA study executive summary and reported in a separate part of the environmental consequences section. Spatial and temporal boundaries and any cumulative impacts addressed within the CIA process should be provided evidence or analysis to support the conclusion. In order to prevent cumulative effects effectively, the focus on CIA should be put on biodiversity, social and economic impacts and global environmental effects. PMID- 11382031 TI - [The transference and distribution of tritium water in the soybean-soil system]. AB - The isotope trace techniques of simulation contamination were applied to explore the rule of transference and distribution of tritium water in a simulated soybeam soil ecosystem. The activity ratio of two forms of tritium in plant and soil sample were measured by 50 days simulation experiment. The results showed that when tritium water was entered the soil, it not only was transferred to other compartments in the system, but also vaporized into atmosphere rapidly. Both free water tritium and bound tritium were found in the soybean and the soil. The activity ratio of free water tritium reduced after reaching the maximum (19.4 Bq.g-1 in root, 12.3 Bq.q-1 in stem and leaf) at 6th hour in the soybean. The bound tritium increased slowly. The activity ratio of total tritium in soybean root was upper than those in stem and leaf in the earlier stage, they reach equipoise each other in the later stage. The activity ratio of two tritium in the surface soil reduced continuously. The compartment model of tracer kinetic was applied to imitate the experimental data. The activity ratio of soil Cs = 88.37e 11.847t + 7.38e-0.030t and the activity ratio of soybean Cb = 10.30(e-0.030t - e 11.847t) were gained. PMID- 11382033 TI - [An incentive mechanism for environment management under unsymmetrical information]. AB - Information unsymmtry is a big obstacle in environment management. In this paper, an incentive mechanism for implementation of effluent charge under unsymmetrical information was designed. This mechanism will provide incentives for true information and lower down the transaction cost in environment management. PMID- 11382034 TI - [Adsorption of anthraquinone dyes by biosorbent GX2]. AB - The GX2 growing mycelium shows excellent adsorptive capacity on the four kinds of anthraquinone dyes, however, due to the difference of molecular structure of dyes, the adsorption rates are different. Although the dyes show inhibiting effect on the growth of GX2, when the concentration of reactive brilliant blue KN R is 250 and 400 mg/L, the adsorption rate of dye by the GX2 growing mycelium reaches 100% and 91.4%, respectively. With the increase of salinity (NaCl), the dry weight of GX2 mycelium increases; the diameter of granular mycelium decreases and its adsorption surface area increases accordingly, the salinity shows evident enhancement on the adsorption of reactive brilliant blue KN-R by GX2 growing mycelium in the range of 0%-2% salinity. The adsorption process is influenced by the concentration of carbon source as well, reactive brilliant blue KN-R with the concentration of 120 mg/L can be completely adsorbed only when the concentration of glucose is more than 2.5 g/L. The GX2 growing mycelium has better adsorptive capacity than resting living mycelium and dead mycelium. PMID- 11382035 TI - [The bindings of typical aldehydes pollutants with cell DNA]. AB - The bindings of 3 kinds of aidehyde pollutants, formaldehyde, acetaldehyde, acrolein, with both prokaryotic and eukaryotic DNA, and their genotoxic effect and mechanism were conducted by the shifts of maximum UV absorption peak to determine binding effects and by HPLC to determine binding sites in vitro model system. The shifts of maximum UV absorption peak of prokaryotic DNA contaminated by 3 kinds of aldehyde pollutants are not significant; but after the DNA extracted from prokaryotic bacteria reacting with formaldehyde in tube, the shifts of maximum UV absorption peak of DNA are significant; the shifts of maximum UV absorption peak of eukaryotic DNA contaminated by 3 kinds of aldehyde pollutants are significant also. The reacition of acetaldehyde with dG reduced by NaBH4 was separated and detected by HPLC, the product was determined qualitatively as N2-ethaldeoxyguanosine adduct. The 3 kinds of aldehyde pollutants could bind with cellular DNA to express genotoxic effects; and the N2 site of deoxyguanosine is the possible covalent binding site. PMID- 11382036 TI - [Toxic effects of Cu, Zn and Mn on the inhibition of Chlorella pyrenoidosa's growth]. AB - A standard method of algal bioassay for evaluating the toxicity of toxic chemicals was applied in the study. It was found that safety concentrations of Cu, Zn and Mn for Chlorella pyrenoidosa's growth were 31.8 micrograms/L, 65.0 micrograms/L and 5.5 mg/L, and 96 h-EC50 were 67.3 micrograms/L, 473.0 micrograms/L and 17.0 mg/L, respectively. According to both safety concentration and 96 h-EC50, the toxic sequences of these metals from high to low on the inhibition of C. pyrenoidosa's growth were Cu, Zn and Mn. The different affinities between different metal ions and algal cells are mainly responsible for the different inhibitions of C. pyrenoidosa's growth. PMID- 11382037 TI - [Effective factors of flocculation-DAF pilot process treating water with low temperature and low turbidity from Miyun reservoir]. AB - The hydraulic parameters in coagulation, flocculation and flotation units of DAF pilot process were systematically investigated when employing flocculants AS and PAC respectively. In general, the available parameters for PAC are more favorable in DAF process. For PAC as a flocculant during DAF run, the favorable coagulation rapid mixing time is 15 s, the flocculant mixing time is no less than 5 minutes. However for AS, the corresponding time values are greater than 30 s and more than 7.5 minutes respectively. The available coagulation mixing intensity is 300-1000 s-1. The available flocculation mixing intensity (average G) is 40-140 s-1, and the GT (average) value is greater than 2 x 10(4). Both equal-speed mixing and two or three stages mixing could provide slight better turbidity removal. For MJ model nozzle, the suitable recycle ratio is 5.0%-8.3% at dispersed pressure of 2.5-3.3 kg.cm2. The detention time and hydraulic intensity in contact zone of DAF tank have important effects on DAF turbidity removal. PMID- 11382038 TI - [Evaluation for the regeneration efficiency of activated carbon by wet air oxidation]. AB - It is well known that different regenerating techniques of activated carbon(AC) require the suitable evaluating methods for regeneration efficiency. In this paper, several methods for evaluating the regeneration efficiency of AC were compared and discussed by wet air oxidation(WAO) which were respectively based on different experimental protocols such as iodine value, molasses color adsorption, breckthrough profile, adsorption isotherm and standard reload experiment. The iodine value, molasses color adsorption and breckthrough profile yielded indefinitely PR values for WAO process, so they could not be applied to evaluate regeneration efficiency. And adsorption isotherm experiment was more correct because it compared isotherm loadings at the same equilibrium liquid phase concentrations, but this more laborious and time-consuming method was difficult to be widely used. The results also showed that the standard reload experiment was consequently picked out, easy to use and had high sensibility alternative to adsorption isotherm experiment. When controlling the WAO parameters at 250 degrees C, 0.6 MPa and 1 h, the regeneration efficiency arrived to 55%. PMID- 11382039 TI - [Ultrafiltration of micropolluted water in combination with coagulation and PAC process]. AB - The experiments of ultrafiltration of Huangpu River water with coagulation and powered activated carbon (PAC) combinations was evaluated. The results showed that coagulation and PAC pretreatment effectively remove dissolved organic matters (DOM), of which coagulation removes high molecular weight (MW) DOM and PAC removes low MW DOM. Coagulation and PAC are effective treatment for removing Trihalomethane Formation Potential (THMFP). In terms of low MW THMEP, coagulation removes poorly and PAC removes effectively. The results also showed that addition of coagulant and PAC reduce cake resistance greatly. At coagulant dosage 4 mg/L (as Al), cake resistance was minimized. PMID- 11382040 TI - [Decomposition of aqueous sodium pentachlorophenolate (PCP-Na) by using TiO2 coating photocatalyst]. AB - Titanium dioxide (TiO2) photocatalysts were coated on hollow glass beads by sodium silicate (Na2SiO3.9H2O) solution. The coated catalysts were characterized by BET, XRD. Their photoactivity were examined using photocatalytic degradation of sodium pentachlorophenolate(PCP-Na) and glucose solution. The impact of catalysts dosage, PCP-Na initial concentration, initial pH and oxygen concentration on reaction kinetics were also studied in detail. The experiments illustrated that the performance of C3-type catalysts was satisfactory. Conditions of the experiments were following: The initial concentration of PCP-Na and CODCr were 10 mg.L-1 and 400 mg.L-1, respectively, and illumination time was 2 hours and catalysts dosage was 2 g.L-1, the removal rates of CODCr and PCP-Na were above 65% and 92% respectively. Optimum dosage of C3-type catalysts was 2 2.5 g.L-1, DO was no less than 3 mg.L-1. PMID- 11382041 TI - [Comparison between porous polymer carrier and activated carbon carrier used for treating organic wastewater in anaerobic fluidized-bed reactor]. AB - A comparative performance between porous polymer carriers (HP) and granular activated carbon carriers (GAC) in anaerobic fluidied-bed reactors was undertaken to evaluate their characters. The results showed that the COD removal and the biogas volume yield rate were 84% and 16.5 m3/(m3.d) respectively when HP was used as carrier to treat synthetic wastewater, at the top COD organic load rate of 65.5 kg/(m3.d), however those were 74.2% and 14.5% respectively for GAC carrier at the top load rate of 63.25 kg/(m3.d). The COD removal and biogas volume yield rate were 64.7%-54.5% and 1.89-2.7 m3/(m3.d) respectively when HP was used as carriers to treat straw pulping wastewater, at the load rate of 14.5 36.15 kg/(m3.d), and those were 61.0%-52.1% and 0.73-2.0 m3/(m3.d) respectively for GAC carriers at the load rate 9.16-19.06 kg/(m3.d). The study revealed that the HP carriers reactor is more efficient than the GAC carriers reactor in microbial immobilization and the wastewater treatment. PMID- 11382042 TI - [The toxicity variation of organic extracts in drinking water treatment processes]. AB - Source water samples and outlet water samples from different treatment processes of the Beijing Ninth Water Works were concentrated in situ with XAD-2 filled columns. GC-MS analysis and toxic assessment including acute toxicity evaluation by luminescent bacterium bioassay(Q67 strains) and mutagenicity assessment by Ames test(TA98 and TA100 strains with and without S9 addition) were conducted on these samples. The results showed that prechlorination caused the direct and indirect frame shift mutagenicity as well as indirect base pair substitute mutagenicity. Addition of coagulant may increase the base pair substitute mutagenic effects greatly. Sand and coal filtration and granular activated carbon filtration could effectively remove most of the formed mutagens. The rechlorination do not obviously increase the mutagenic effects. No mutagenic effect was observed in tap water. Acute toxicity showed the same variation with that of mutagenicity during the treatment processes. Sample from flocculation treatment process was found to be the most toxic sample. Results of GC-MS analysis showed that water in this plant was not contaminated by PCB. Concentrations of toluene, naphthalene and phenol increased in flocculation treatment process and in tap water. However, the concentrations of these substances were at the level of microgram/L, therefore, were not high enough to cause mutagenicity. PMID- 11382043 TI - [Hydrodynamic property of internal airlift loop bioreactor with cells immobilized onto ceramic honeycomb support(IALBR-CICHS)]. AB - An internal airlift loop biorector with cells immobilized onto ceramic honeycomb support(IALBR-CICHS) was developed based on the bubble columns(BC) and airlift(AL) bioreactor by installing a ceramic honeycomb support in the draught tube. Their hydrodynamics were respectively studied by the tracer element to determine their retention time distribution (RTD). Experiment and theory analysis indicated that the IALBR-CICHS had more efficacious reactive volume rate(eta) and less short-circuit rate(lambda) for fluid flow compared with the BC and AL bioreactor. PMID- 11382045 TI - [Mass concentration and major inorganic compositions of coastal aerosol in Qingdao]. AB - Five times measurements of atmosphere aerosol were conducted from September, 1997 to November, 1998 at a coastal site of Qingdao. The mass concentrations of aerosol and their main soluble compositions were measured. The mass concentration and seasonal variation of the aerosol and its composition were discussed by different size. The PM2.5 (d < 2.5 microns) mass concentration accounts for half of the TSP, while the PM10 (d < 10 microns) mass concentration is about 75% of the TSP. More than 40% of the TSP mass concentration are from that of the soluble ions, among which SO4(2-), NO3- and NH4+ are the primary ones. SO4(2-), NH4+, NO3 are mainly in fine particles which suggest that they mainly existed in secondary aerosols. The mass concentrations of TSP in winter are much higher than those in other seasons. PMID- 11382044 TI - [Effect of LAS on physico-chemical properties and microorganism activity in soil environment]. AB - Using a well drained paddy soil and the derived soil under vegetables in the Taihu Lake region, China, experiments of soil capillary rise, colloid dispersion and adsorption of toxic organics and heavy metals by soil under various degrees of LAS addition were conducted. Also incubation experiments of microorganism activity in soil with added LAS in different concentrations were carried out in laboratory. LAS addition resulted in lower capillary rise of soil solution, higher dispersion of soil colloids and lower capacity of soil for organic pollutants but no significant influence on adsorption of heavy metal cations. The nitrification and amonification in soil were very sensitive to LAS stress while denitrification turned to be stimulated. LAS addition gave stronger growth of soil bacteria while decreased growth of fungus. Meanwhile, soil respiration was stimulated in the first week of incubation and compressed afterwards by LAS at concentration over 5 mg/kg. It seemed that some species of microorganisms were tolerant of LAS in the studied soils. PMID- 11382046 TI - [Combustion characteristics of municipal solid waste in fluidized bed]. AB - For the experimental investigation on the combustion characteristics of Municipal Solid Waste(MSW) in Fluidized Bed, a lab scale Fluidized Bed facility was constructed. Single and mixed municipal solid waste burning in Fluidized Bed showed that dry waste can be burnt quickly at the bed temperature of 500 degrees C, and furnace temperature increased about 30-50 degrees C. Many kinds of combustion runs were conducted in this Fluidized Bed combustion facility. The parameters examined were air flow rate(from 5.5 m3/h to 7.5 m3/h), form of fuels(scrap or whole), moisture content and so on. Concentration of CO2, CO, SO2 and NOx in the flue gas were monitored and recorded every 5 seconds. The temperatures along the reactor were recorded every 10 seconds. PMID- 11382047 TI - [Chemical assessment of mutagenicity of halohydrocarbons: FTIR, UV spectrum characteristics of guanine-halohydrocarbon adducts]. AB - Determination and identification of guanine-halohydrocarbon adduct isomers were carried out by high performance liquid chromatography (HPLC) and Fourier transform infrared spectroscopy (FTIR). The experimental results showed that the UV spectral overlay of O6-guanine adducts retrieved after HPLC at 260-270 nm. The UV spectra of N-side adducts are similarly between 240-250 nm. After thin layer chromatographic separation (silica gel-GF254), the all IR spectra of the N-side adducts of guanine with different halohydrocarbons showed a absorption near 1700 cm-1 assigned the characteristic peak of C=O. The IR spectra among the adducts of O6-guanine with different halohydrocarbons are quite similar. PMID- 11382048 TI - [Treatment of dilute solution anilines-containing with displacement by extraction]. AB - It was found that the biodegradability (indicated by BOD5/CODCr value of water solution containing anilire or chloroaniline declined with the increasing of their concentration. In order to bio-treat such water, decrease of the solute's concentration was necessary. Based on the research of organic solvents' and complexing agent's biodegradability, a series of extraction experiments were conducted for aniline and m-chloroaniline with complexing agent or physical solvent. The factors influencing the distribution ratio were discussed. The BOD5/CODCr value of raffinates showed that the raffinate can be treated by biological degradation without dilution if suitable solvent was choosed. It was proved that displacement by extraction had great potentiality for treating wastewater containing non-biodegradable organic contaminant. PMID- 11382049 TI - [Removal of TOC and Ames mutagens from drinking water using AC-NF process]. AB - Expermental results of the effect and mechanism of removing TOC and Ames mutagens from drinking water using AC-NF process showed that the adsorptive ability of AC was limited, depending on the characters of AC and the organic matters in drinking water. But AC can be used as the pretreatment of NF to ensure to meet the requirement of membrane feed. NF can remove the majority of total organic carbon and Ames mutagens effectively to ensure the safety of drinking water. The MR values of TA98 and TA100 of NF permeate were below 2 under the tested doses, and the results of Ames test of NF permeates were completely negative. Therefore, it is feasible to combine AC and NF to obtain the high-grade drinking water. PMID- 11382050 TI - [Quinoline removal in fluidized-bed bioreactor using immobilized cells]. AB - Burkholderia pickettii, capable of using quinoline as sole source of carbon, nitrogen and energy, was immobilized by an innovative PVA-H3BO3-gauze method. A fluidized-bed bioreactor, in which immobilized cells were added, was constructed to degrade quinoline in wastewater. When the initial concentrations of quinoline were 100, 350 and 500 mg/L, the bioreactor can remove it completely within 2.5, 6 and 12 hours respectively. A series of zero-order reaction equations were proposed to describe quinoline biodegradation process in this bioreactor kinetically. In continuous flow experiments, the performance of the bioreactor was investigated at different HRT (dilution rate) for different quinoline concentrations in wastewater and the tolerance against shock loading was determined by increasing quinoline concentration from 100 to 300 to 500 mg/L at HRT 4 hours. PMID- 11382051 TI - [Vertical distribution characteristics of organochlorinated pesticides in sediment core from Macao estuary, Pearl River delta]. AB - This work presents the analysis result of concentration for organochlorinated pesticides in a sedimentary core sample. Pesticides were quantified by GC/ECD with the internal standard material. The core dated by 210Pb methods. The vertical distribution of pesticides concentration with the depth was discussed. The results showed the concentration of BHCs and DDTs in the core ranged from 0.48 ng/g to 26.28 ng/g and 1.92-39.13 ng/g respectively. The distributing characteristics of organochlorinated pesticides in the core varied with the water fluxes of Pearl River. Organochlorinated pesticides may come from the residue of agricultural soils. Concentration of DDTs was above the effects range-low value for marine and estuarine sediments. PMID- 11382052 TI - [The survey of polycyclic aromatic hydrocarbons in a habitation air in Hangzhou]. AB - The twelve polycyclic aromatic hydrocarbons(PAHs) were detected in a habitation air air of Hangzhou. The results indicated that the concentrations of PAHs in the habitation air were between 8.237 micrograms/m3 and 14.234 micrograms/m3. The habitation air was seriously polluted by PAHs. The main source of PAHs in the habitation air was from cooking and burning. And the waste gases from vehicular will contribute to it also. PMID- 11382053 TI - [Sediment-copper interaction and vertical distribution in hyper-concentrated turbulent system]. AB - This paper presents a special problem on vertical distribution for sediment and copper in hyper-concentrated turbulent solid-liquid system that is essentially different from the ordinary low concentrated turbulent system. A resonance type turbulent simulation equipment is used for the experimental study in which a vertically uniform turbulent field of the mixture of loess and water is produced in a testing cylinder with a grille stirrer that moves up and down harmoniously with varying vibration frequencies. In order to compare the variations of the vertical profiles of sediment and copper in low- and hyper-concentrated solid liquid system, different scenarios for input sediment content ranging from 5 to 800 kg/m3 was considered in the experimental studies. It was found that solids copper content increases with input sediment content, S, and reaches its peak as S goes to 10 kg/m3 and then decreases rapidly with increasing input sediment content. Such a behavior is possibly resulted from the joint effect of the specific adsorption of copper on loess, precipitation of carbonate and hydroxide of copper due to high carbonate content in the loess and the so-called "particulate concentration effect" due to the present of the sediment variation in water. The vertical sediment concentration distribution resulted from the uniform turbulence is generally uniform, but slight non-uniformity does occur as sediment concentration exceeds certain value. However, the vertical concentration distributions of soluble copper seem not affected much by the variation of sediment concentrations. PMID- 11382054 TI - [Characterization of speciation distribution of IPF-PFSi]. AB - The preparation and speciation distribution characteristic of a new kind of inorganic polymer flocculant(IPF)-PFSi were investigated in detail by the combination of direct and indirect speciation analysis methods. The experimental results show that the speciation distribution of PFSi is decided significantly with basicity, kinds of silica and Si/Fe ratio. Among them, the basicity is one of the main factors. SilicaA and silicaB exhibit a similar effect, while silicaC has limited effect on the hydrolysis of Fe(III). The typical speciation distribution and transformation property of PFCl changes limitedly with the introduction of silica species. PMID- 11382055 TI - [Phosphorus loss potential of soil-water in sites of the main rice-yield area in the Northern Zhejiang]. AB - Phosphorus (P) loss potential and its environmental impact from soil-water were surveyed in Jiashan Yuyao, Deqing and Yuhang, named main rice-yield areas in Northern Zhejiang province China. High P input has resulted in soil P accumulation. Higher soil Olsen-P contributed its bio-available P, water extracted P and increased soil P loss potential. The role of P in paddy soils is of environmental rather than agronomic concern in the process of soil P build-up. During the no-rice time, total P (TP) concentration in the various water including ditch drain inside/outside field, field surface water and pipe drain et al are over the critical values associated with accelerated waters eutrophication. The average proportion of dissolved reactive P (DRP) concentration in TP was 40%, however, these current DRP levels in partly ditch drain outside field and surface or ground water in no-rice time could not caused serious impact to the outside waters nearby rice-yield agriculture area. No significant line correlation between soil P tests and P in field surface water was found in no-rice time due to differences in field tillage among rice-yield areas, while P application increased both soil P and its correspondingly field surface P level after rice was planted at the same time. PMID- 11382056 TI - Evaluation of individual components of plum odor as potential attractants for adult plum curculios. AB - We evaluated olfactory attraction of overwintered plum curculio (PC) adults, Conotrachelus nenuphar, to 16 individual volatile components of unripe plum odor in the laboratory using a still-air dual-choice bioassay system and in the field using baited cotton dental wicks attached to boll-weevil traps placed on the ground beneath the canopy of unsprayed apple trees. Two compounds, ethyl isovalerate and limonene, were significantly attractive in both laboratory bioassays and field experiments. In laboratory bioassays, as concentration was decreased across five orders of magnitude, a greater number of compounds elicited responses suggestive of attractancy (except at the lowest concentration). Even so, linalool, 2-hexanone, and 3-hydroxy-2-butanone were the only other compounds showing significant attractiveness in laboratory bioassays, but none of these (nor any other compounds) were significantly attractive in field assays. We suggest that the use of ethyl isovalerate and/or limonene as odor attractants offers potential to increase the efficacy of current traps for monitoring PCs immigrating into fruit orchards during spring. PMID- 11382057 TI - Mandibular gland chemistry of grass-cutting ants: species, caste, and colony variation. AB - The compositions of the alarm pheromones of two species of grass-cutting ant, Atta bisphaerica and A. capiguara, were examined, and caste and colony variations quantified. The pheromones of A. bisphaerica and A. capiguara were remarkably similar and were composed of a complex mixture of volatiles in which 4-methyl-3 heptanone and 2-heptanone were the most abundant compounds. Small but consistent intraspecific differences were found between the worker castes and between individual colonies. The results support the view that alarm pheromones are rarely species specific. The possible importance of intercolony variation is discussed. PMID- 11382058 TI - Allelochemicals in wheat (Triticum aestivum L.): variation of phenolic acids in shoot tissues. AB - Seven known phenolic acids implicated in wheat allelopathy were analyzed in a worldwide collection of 58 wheat accessions by gas chromatography and tandem mass spectrometry (GC-MS-MS). Chemical analysis showed that accessions differed significantly in the production of p-hydroxybenzoic, vanillic, syringic, trans-p coumaric, cis-p-coumaric, trans-ferulic, and cis-ferulic acids in the shoots of 17-day-old wheat seedlings. The concentrations of p-hydroxybenzoic, vanillic, cis p-coumaric, and cis-ferulic acids were normally distributed in the 58 accessions. A binormal distribution was found for syringic and trans-ferulic acids and a skewed normal distribution for trans-p-coumaric acid. The concentration of each compound also varied with phenolic acids. The relative abundance of each phenolic acid was ordered decreasingly as trans-ferulic, vanillic, trans-p-coumaric, p hydroxybenzoic, syringic, cis-ferulic, and cis-p-coumaric acids. The concentration of total identified phenolic acids varied from 93.2 to 453.8 mg/kg in the shoots of 58 accessions. The content of each phenolic acid or group was highly associated with others in the shoots of wheat seedlings. Wheat accessions with high levels of total identified phenolic acids in the shoots are generally strongly allelopathic to the growth of annual ryegrass. PMID- 11382059 TI - Plant-growth inhibitory activity of cedrelanolide from Cedrela salvadorensis. AB - The effect of cedrelanolide, the most abundant limonoid isolated from Cedrela salvadorensis (Meliaceae), was assayed as a plant-growth inhibitory compound against monocotyledonous and dicotyledonous seeds. This compound inhibited germination, seed respiration, and seedling dry weights of some plant species (Lolium multiflorum, var. Hercules, Triticum vulgare, var. Salamanca, Physalis ixocarpa, and Trifolium alexandrinum). Our results indicate that cedrelanolide interferes with monocot preemergence properties, mainly energy metabolism of the seeds at the level of respiration. In addition, the compound inhibits photophosphorylation, H+ uptake, and noncyclic electron flow. This behavior might be responsible for its plant-growth inhibitory properties and its possible role as an allelopathic agent. PMID- 11382060 TI - Influence of defoliation on toxic alkaloid concentration and alkaloid pools in tall larkspur. AB - This study was replicated at two locations in the mountains of central Utah. In 1997, ten uniform plants of tall larkspur (Delphinium barbeyi) in the early bud stage (40 cm in height) were selected at each site and clipped at 5 cm above soil level. In 1998, one stalk from each plant was harvested on a weekly basis; in 1999, one stalk was harvested at four times during its phenological development. Toxic and total alkaloid concentrations were measured and alkaloid pools in the entire stalk were calculated. Clipping reduced stalk height to less than 50 cm in 1998 and 65 cm in 1999, compared to over 100 cm in unclipped control plants. Alkaloid concentration was similar to control plants, but toxic alkaloid pools were 70% lower than control plants, because of the reduction in biomass of the stalks. Clipping reduced subsequent vigor and the amount of toxic and total alkaloids in tall larkspur. PMID- 11382061 TI - Involvement of ligninlike compounds in toxicity of dietary alder leaf litter against mosquito larvae. AB - The toxicological characteristics of dietary decomposed alder leaf litter against mosquito larvae were further investigated through enzymatic and chemical purification of a phenoliclike cell-wall fraction isolated from crude litter. The toxicity of the subfractions obtained was controlled by standard bioassays on third instars of Aedes aegypti chosen as a reference target species. Enzymatic hydrolyses of the cell-wall fraction were performed with caylase, pectolyase, esterase, and beta-glycosidase, in order to release, respectively, cellulose material and phenolic compounds bound to lignins. These treatments did not affect the larvicidal activity and the phenolic activity of the cell-wall fraction. Chemical alkaline and acid hydrolyses were carried out to break ester and glycosidic bonds of the cell-wall fraction. Comparison of HPLC profiles of the hydrolysates from both toxic and nontoxic fractions did not reveal differences between the phenolic acids released. Aluminum chloride, known for its phenolic complexing activity, counteracted the larvicidal activity of the cell-wall fraction. Altogether, these results suggest the involvement of ligninlike compounds in the toxicity of dietary alder leaf litter against larval mosquitoes. The toxicity of this fraction, which was very sensitive to drastic and smooth oxidations, seemed to be associated with a strong oxidative potential. These results are discussed in relation to a possible mode of action of lignins in the plant-mosquito interactions. PMID- 11382062 TI - Thin-layer chromatography assessing feeding stimulation by labial gland secretion compared to synthetic chemicals in the subterranean termite Reticulitermes santonensis. AB - The labial gland of the French subterranean termite Reticulitermes santonensis De Feytaud contains a polar, heat-resistent, and persistent chemical signal that is released onto the food during food exploitation and stimulates feeding in nestmates. Separation of the labial gland secretion by thin-layer chromatography on cellulose plates revealed that the secretion contains components with reducing and amino groups. In feeding bioassays conducted on the cellulose plate after TLC, termites preferred the area between Rf 0.46 and 0.88 (biologically active zone) for feeding, indicating the location of the feeding-stimulating signal. Thirty-five synthetic chemicals with similar chemical properties as the feeding stimulating signal were analyzed with TLC. None of them covered the biologically active zone. Therefore, all chemical classes tested, such as sugars, amino acids, and salts, are unlikely as possible sources for the signal structure. In feeding choice tests with synthetic chemicals, termites showed clear feeding preference only for sugarlike components with physiologically excessive concentrations of 10 mmol and 100 mmol. Amino acids induced only light feeding preference. The intensity of feeding stimulation by the natural signal from the labial gland as compared to synthetic phagostimulants is discussed. PMID- 11382063 TI - Food exploitation in termites: indication for a general feeding-stimulating signal in labial gland secretion of isoptera. AB - The paired labial glands are located in all termite species in the thorax. During food exploitation workers of the French termite Reticulitermes santonensis and the African termite Schedorhinotermes lamanianus release the secretion of their labial glands directly onto the food. The secretion carries a water-soluble, heat resistant, nonvolatile signal that stimulates gnawing and feeding and leads to aggregations of feeding workers. In a feeding bioassay, extracts of the labial glands of 11 termite species from five families all proved to have this feeding stimulating effect both on R. santonensis and S. lamanianus. The heat resistance of the feeding-stimulating signal also could be shown for selected species from all termite families tested. A combined thin layer chromatography-feeding bioassay on cellulose TLC plates showed that after chromatography of labial gland secretion, the feeding-stimulating signal is located in all 11 species in the same area from Rf 0.46 to 0.88. An extract of labial glands of cockroaches stimulated feeding in R. santonensis and S. lamanianus as well, but was not active after heat treatment and after TLC. This points towards a general feeding stimulating signal having evolved only in the labial gland secretion of termites. PMID- 11382064 TI - Plant growth inhibitory activity of L-canavanine and its mode of action. AB - L-Canavanine is a nonprotein amino acid contained in jack bean [Canavalia ensiformis (L.) DC] and shows a plant inhibitory effect. The inhibitory effect was determined by an immersion test and a microdrop test that employed rice seedlings. L-Canavanine inhibited elongation of the second leaf sheath of rice seedlings more than other natural bioactive substances, such as salicylic acid and cinnamic acid. The modified microdrop test revealed that the mode of action of L-canavanine had no relation to gibberellin synthesis. In the microdrop test, the inhibitory effect of L-canavanine was decreased by simultaneous addition of L arginine, an analog of L-canavanine. Free amino acid analysis of rice shoots clearly showed that L-canavanine induced an unusual accumulation of L-arginine. However, accumulation of L-arginine did not cause the inhibitory effect on plant growth. These results suggest that the mechanism of inhibition of L-canavanine is closely related to the inhibition of arginine metabolism. PMID- 11382065 TI - Response of the ladybird parasitoid Dinocampus coccinellae to toxic alkaloids from the seven-spot ladybird, Coccinella septempunctata. AB - Electrophysiological and behavioral responses of the ladybird parasitoid Dinocampus coccinellae to volatiles from the seven-spot ladybird, Coccinella septempunctata, were investigated to identify semiochemicals involved in host location. Coupled gas chromatography-electroantennography (GC-EAG) with D. coccinellae located a small peak of prominent activity in an extract of volatiles from adult C. septempunctata. The active compound was identified by coupled GC mass spectrometry and by comparison with an authentic sample as the free-base alkaloid precoccinelline, which forms part of the toxic defense of this ladybird. Behavioral studies in an olfactometer showed that D. coccinellae was significantly attracted to the volatile extract and also to the alkaloid. Myrrhine, a stereoisomer of precoccinelline found in low amounts in C. septempunctata and in other ladybird species, was shown to be electrophysiologically active and significantly attractive. Perception of ladybird alkaloids by D. coccinellae is a rare example of toxicants acting as aerially transmitted cues for interactions between the third and fourth trophic levels. PMID- 11382066 TI - Importance of predator diet cues in responses of larval wood frogs to fish and invertebrate predators. AB - We examined the effects of predator diet on the antipredator responses of larval woodfrogs (Rana sylvatica). We found that tadpoles showed stronger responses to fish (Perca flavescens) that were fed tadpoles than those fed invertebrates. Similarly, we found that tadpoles responded more strongly to larval dragonflies (Anax spp.) fed tadpoles than to dragonflies fed invertebrates. The overall intensity of response of tadpoles to fish was much stronger than that to dragonflies. Predator diet effects are not ubiquitous in predator-prey systems. We discuss possible reasons why predator diet effects are seen in some, but not all, predator-prey systems. PMID- 11382067 TI - Ethylene production and peroxidase activity in aphid-infested barley. AB - The purpose of this work was to investigate whether ethylene is involved in the oxidative and defensive responses of barley to the aphids Schizaphis graminum (biotype C) and Rhopalophum padi. The effect of aphid infestation on ethylene production was measured in two barley cultivars (Frontera and Aramir) that differ in their susceptibility to aphids. Ethylene evolution was higher in plants infested for 16 hr than in plants infested for 4 hr in both cultivars. Under aphid infestation, the production of ethylene was higher in cv. Frontera than in Aramir, the more aphid susceptible cultivar. Ethylene production also increases with the degree of infestation. Maximum ethylene evolution was detected after 16 hr when plants were infested with 10 or more aphids. Comparing the two species of aphids, Schizaphis graminum induced more ethylene evolution than Rhopalosiphum padi. Infestation with S. graminum increased hydrogen peroxide content and total soluble peroxidase activity in cv. Frontera, with a maximum level of H2O2 observed after 20 min of infestation and the maximum in soluble peroxidase activity after 30 min of infestation. When noninfested barley seedlings from cv. Frontera were exposed to ethylene, an increase in hydrogen peroxide and in total peroxidase activity was detected at levels similar to those of infested plants from cv. Frontera. When noninfested plants were treated with 40 ppm of ethylene, the maximum levels of H2O2 and soluble peroxidase activity were at 10 and 40 min, respectively. Ethylene also increased the activity of both cell-wall-bound peroxidases types (ionically and covalently bound), comparable with infestation. These results suggest that ethylene is involved in the oxidative responses of barley plants induced by infestation. PMID- 11382068 TI - Secretions of stingless bees: the Dufour gland of Nannotrigona testaceicornis. AB - The Dufour gland of Nannotrigona testaceicornis is a large, wide, pear-shaped sac. The gland secretion consists chiefly of the diterpene ester all-trans geranylgeranyl acetate (64% of the total), together with a complex mixture of small amounts of cyclic ketals; mono-, sesqui-, and diterpene compounds; acetates; and other oxygenated compounds. Samples of N. testaceicornis collected at two sites in Brazil and one in Mexico shared the same composition of their glands, suggesting that the species is uniform over this wide geographical area. PMID- 11382069 TI - Phenolic compounds in a Sahelian sorghum (Sorghum bicolor) genotype (CE145-66) and associated soils. AB - CE145-66 is an improved early-maturing grain sorghum genotype, increasingly grown by farmers in the Sahelian part of Senegal. This genotype is known to have negative effects on the following groundnut crop, because of the release of allelopathic phenolic compounds into the soil. We have assessed the synthesis of phenolics in sorghum vegetative parts and the variations in synthesis between years and sites. Total phenols and phenolic acids in the aerial parts and roots of flowering sorghum plants from 52 farmers' fields at two sites (Sagnaneme and Medina) in Senegal in 1996 and 1997 were measured. Thirty-eight soil samples, collected after the sorghum harvest, from sorghum rows and interrows also were analyzed for their phenolic content. Total phenols reached 1.1-1.5% of root dry weight and 1.1-2.2% of aerial parts dry weight, with little variation between sites, and large variability between years, presumably due to climatic conditions. Eight phenolic acids and three associated aldehydes were identified by HPLC, with p-hydroxybenzoic, p-coumaric, and ferulic acids the most abundant. Their totals reached 2.9-3.2 mg/g in 1996 and 2.6-2.8 mg/g in 1997 for the aerial part; and 3.3-3.6 mg/g in 1996 and 2.8-3.3 mg/g in 1997 for roots. In soils under sorghum rows, the mean water-soluble total phenols increased from 4.6 in 1997 to 6.7 micrograms/g in 1998 in Sagnaneme, and from 3.8 in 1997 to 5 micrograms/g in 1998 in Medina. The concentrations of total phenols and phenolic acids were higher in rows than in interrows. All the phenolic monomers identified in vegetative parts were recovered in associated soil samples, with vanillic and p hydroxybenzoic acids the most abundant. Finally, variability in plant phenolic content seemed more due to climatic than to cropping or soil factors, as differences between years appear more important than differences between or within sites. PMID- 11382070 TI - Chemical ecological characteristics of herbivory of Siparuna guianensis seeds by buffy-headed marmosets (Callithrix flaviceps) in the Atlantic Forest of southeastern Brazil. AB - The buffy-headed marmoset (Callithrix flaviceps) is apparently the only predispersal herbivore of the seeds of Siparuna guianensis at the Caratinga Biological Station in southeastern Brazil. Both the fruit receptacles and the frutioles (seeds) of S. guianensis are relatively rich in nutrients such as carbohydrates, proteins, and lipids, but the receptacles contain high concentrations of benzylisoquinoline alkaloids. The latter presumably act as a qualitative chemical defense, impeding the access of potential predators to the alkaloid-poor frutioles. However, on ripening, the receptacle splits open, exposing the frutioles, which enables C. flaviceps to avoid the plant's chemical defenses effectively. Taking care to avoid contact with the receptacle, the marmosets pluck out the frutioles and ingest the seeds. Qualitative and quantitative changes (in particular a significant reduction in daily ranging) in the marmosets' behavior during the period when S. guianensis frutioles were accessible indicate that this was a preferred plant resource. This is the first record of the consumption of S. guianensis seeds by callitrichine monkeys (which are not known to be systematic seed eaters), despite the fact that both are widely distributed in the Neotropics. It is thus possible that the behavioral strategy observed here is a unique phenomenon resulting from a specific combination of factors, including the abundance of S. guianensis within the study area. The lack of other records may nevertheless be a result of insufficient sampling effort, in which case, the feeding strategies of callitrichines may have an important influence on the reproductive biology of S. guianensis in many areas. PMID- 11382071 TI - A novel method for the prediction of functional biological activity of polyethylene wear debris. AB - The comparative performance of artificial hip joints has been extensively investigated in vitro through measurements of wear volumes. In vivo a major cause of long-term failure is wear-debris-induced osteolysis. These adverse biological reactions are not simply dependent on wear volume, but are also controlled by the size and volumetric concentration of the debris. A novel model is presented which predicts functional biological activity; this is determined by integrating the product of the biological activity function and the volumetric concentration function with the wear volume over the whole particle size range. This model combines conventional wear volume measurements with particle analysis and the output from in vitro cell culture studies to provide a new indicator of osteolytic potential. The application of the model is demonstrated through comparison of the functional biological activity of wear debris from polyethylene acetabular cups articulating under three different conditions in a hip joint simulator. PMID- 11382072 TI - Effect of contact stress on friction and wear of ultra-high molecular weight polyethylene in total hip replacement. AB - This paper studies the effect of contact stress on friction and wear of ultra high molecular weight polyethylene (UHMWPE) acetabular cups by means of friction and wear joint simulator testing under serum lubrication. For a given applied load, increasing the contact stress by increasing the ball/socket radial clearance decreased both the coefficient of friction and the wear rate. Friction and wear were highly correlated. The dependence of friction on contact stress for the UHMWPE socket under serum lubrication was similar to that of semi-crystalline polymers under dry sliding. This finding indicates the occurrence of partial dry contact at asperity levels for the metal-polyethylene ball-in-socket joint under serum lubrication. PMID- 11382073 TI - Elastohydrodynamic lubrication analysis of ultra-high molecular weight polyethylene hip joint replacements under squeeze-film motion. AB - Elastohydrodynamic lubrication was analysed under squeeze-film or normal approach motion for artificial hip joint replacements consisting of an ultra-high molecular weight polyethylene (UHMWPE) acetabular cup and a metallic or ceramic femoral head. A simple ball-in-socket configuration was adopted to represent the hip prosthesis for the lubrication analysis. Both the Reynolds equation and the elasticity equations were solved simultaneously for the lubricant film thickness and hydrodynamic pressure distribution as a function of the squeeze-film time was solved using the Newton-Raphson method. The elastic deformation of the UHMWPE cup was calculated by both the finite element method and a simple equation based upon the constrained column model. Good agreement of the predicted film thickness and pressure distribution was found between these two methods. A simple analytical method based upon the Grubin-Ertel-type approximation developed by Higginson in 1978 [1] was also applied to the present squeeze-film lubrication problem. The predicted squeeze-film thickness from this simple method was found to be remarkably close to that from the full numerical solution. The main design parameters were the femoral head radius, the radial clearance between the femoral head and the acetabular cup, and the thickness and elastic modulus for the UHMWPE cup; the effects of these parameters on the squeeze-film thickness generated in current hip prostheses were investigated. PMID- 11382074 TI - In vitro comparison of the two hard-hard articulations for total hip replacements. AB - Polyethylene particle disease is one of the major causes of late aseptic loosening of total hip replacement. Two hard-hard articulations (alumina-on alumina and metal-on-metal) have been developed in Europe as an alternative to the ultra-high molecular weight polyethylene (UHMWPE) articulations. Even though these hard-hard articulations are on the market and numerous reports have been published about them, only a very limited number of studies allowing a direct in vitro comparison of the two articulations have been published so far. This paper compares in vitro these two types of articulation (alumina-on-alumina and metal on-metal), which have been tested with a hip simulator for their tribological behaviour using exactly the same experimental methodology. This comparison shows that these two types of hard-hard articulation have very similar abrasive wear behaviour with four main features: 1. A running-in wear period (1 x 10(6) cycles) gives a cumulative wear of about 20 microns with head diameters of 28 mm. 2. After the running-in wear, there is a stabilization of the linear wear behaviour with a low linear wear rate/10(6) cycles for both types of articulation. 3. The volumetric wear rate of both articulations (< 2.0 mm3/year for head diameters of 28 mm) is significantly lower than that observed for metal-on-polyethylene or ceramic-on-polyethylene articulations having the same head diameter. 4. Abrasive wear is readily apparent (indicating a mixed lubrication regime) with both types of articulation. The extremely low wear performance of these articulations is confirmed and they constitute a low-wear alternative to the UHMWPE articulations currently used. PMID- 11382075 TI - The effect of femoral head diameter upon lubrication and wear of metal-on-metal total hip replacements. AB - It has been found that a remarkable reduction in the wear of metal-on-metal hip joints can be achieved by simply increasing the diameter of the joint. A tribological evaluation of metal-on-metal joints of 16, 22.225, 28 and 36 mm diameter was conducted in 25 per cent bovine serum using a hip joint simulator. The joints were subject to dynamic motion and loading cycles simulating walking for both lubrication and wear studies. For each size of joint in the lubrication study, an electrical resistivity technique was used to detect the extent of surface separation through a complete walking cycle. Wear of each size of joint was measured gravimetrically in wear tests of at least 2 x 10(6) cycles duration. Joints of 16 and 22.225 mm diameter showed no surface separation in the lubrication study. This suggested that wear would be proportional to the sliding distance and hence joint size in this boundary lubrication regime. A 28 mm diameter joint showed only limited evidence of surface separation suggesting that these joints were operating in a mixed lubrication regime. A 36 mm diameter joint showed surface separation for considerable parts of each walking cycle and hence evidence of the formation of a protective lubricating film. Wear testing of 16 and 22.225 mm diameter metal-on-metal joints gave mean wear rates of 4.85 and 6.30 mm3/10(6) cycles respectively. The ratio of these wear rates, 0.77, is approximately the same as the joint diameters ratio, 16/22.225 or 0.72, as expected from simple wear theory for dry or boundary lubrication conditions. No bedding-in was observed with these smaller diameter joints. For the 28 mm diameter joint, from 0 to 2 x 10(6) cycles, the mean wear rate was 1.62 mm3/10(6) cycles as the joints bedded-in. Following bedding-in, from 2.0 x 10(6) to 4.7 x 10(6) cycles, the wear rate was 0.54 mm3/10(6) cycles. As reported previously by Goldsmith et al. in 2000 [1], the mean steady state wear rate of the 36 mm diameter joints was lower than those of all the other diameters at 0.07 mm3/10(6) cycles. For a range of joints of various diameters, subjected to identical test conditions, mean wear rates differed by almost two orders of magnitude. This study has demonstrated that the application of sound tribological principles to prosthetic design can reduce the wear of metal-on-metal joints, using currently available materials, to a negligible level. PMID- 11382076 TI - Quantification of third body damage to the tibial counterface in mobile bearing knees. AB - Fourteen pairs of explanted low contact stress (LCS) tibial interface components: six rotating platform (RP), six meniscal (MN) and two anterior-posterior (AP) glide designs, have been analysed with particular attention paid to the condition of the tibial counterfaces. The average surface roughness, Ra, for the tibial trays ranged from 0.01 to 0.087 micron, significantly greater than the unworn control measurement of 0.008 micron. The scratch geometry analysis showed that the scratch peaks were found to be consistently of a lower aspect ratio than the scratch valleys and under 1 micron in height (average asperity height Rp = 0.52 micron, aspect ratio delta p = 0.01, average asperity depth Rv = 1.10 microns, delta v = 0.05). The largest scratches were 3-4 microns in both Rp and Rv. In vitro tests have shown that ultra-high molecular weight polyethylene (UHMWPE) wear increases in the presence of counterface scratches perpendicular to the direction of motion. In these explants, the unidirectional motion produced scratches parallel to the direction of sliding which is predicted to produce a smaller increase in UHMWPE wear. Other designs in mobile bearing knees have less constrained motion at the tibial counterface and this has been shown to accelerate wear; it may also lead to a further increase in wear in the presence of third body scratches. It may be possible in future knee designs to reduce this type of wear damage by introducing alternative materials or coatings which are more resistant to scratching and surface roughening. PMID- 11382077 TI - Computational studies of 'whiplash' injuries. AB - The term 'whiplash' was initially used to describe injuries to the neck caused by the head being forced backwards during a rear-end collision in cars without head restraints. The addition of head restraints in the 1970s was expected to solve this problem by preventing excessive extension of the neck but experience suggests the problem still exists. This paper reviews available experimental studies of whiplash and uses the data to construct a finite element model which is capable of dynamically simulating whiplash collisions and predicting the forces in all the relevant neck ligaments. For the first time, it is shown that trauma occurs long before the head hits the head restraint as a result of displacement between the head and the torso caused by the head's inertia leading to markedly different acceleration histories. It is concluded that experimental and computational studies must be used together to produce progress in biomechanical studies. PMID- 11382078 TI - Fabrication of porous polymeric matrix drug delivery devices using the selective laser sintering technique. AB - New techniques in solid freeform fabrication (SFF) have prompted research into methods of manufacturing and controlling porosity. The strategy of this research is to integrate computer aided design (CAD) and the SFF technique of selective laser sintering (SLS) to fabricate porous polymeric matrix drug delivery devices (DDDs). This study focuses on the control of the porosity of a matrix by manipulating the SLS process parameters of laser beam power and scan speed. Methylene blue dye is used as a drug model to infiltrate the matrices via a degassing method; visual inspection of dye penetration into the matrices is carried out. Most notably, the laser power matrices show a two-stage penetration process. The matrices are sectioned along the XZ planes and viewed under scanning electron microscope (SEM). The morphologies of the samples reveal a general increase in channel widths as laser power decreases and scan speed increases. The fractional release profiles of the matrices are determined by allowing the dye to diffuse out in vitro within a controlled environment. The results show that laser power and scan speed matrices deliver the dye for 8-9 days and have an evenly distributed profile. Mercury porosimetry is used to analyse the porosity of the matrices. Laser power matrices show a linear relationship between porosity and variation in parameter values. However, the same relationship for scan speed matrices turns out to be rather inconsistent. Relationships between the SLS parameters and the experimental results are developed using the fractional release rate equation for the infinite slab porous matrix DDD as a basis for correlation. PMID- 11382079 TI - A three-dimensional finite element model from computed tomography data: a semi automated method. AB - Three-dimensional finite element analysis is one of the best ways to assess stress and strain distributions in complex bone structures. However, accuracy in the results may be achieved only when accurate input information is given. A semi automated method to generate a finite element (FE) model using data retrieved from computed tomography (CT) was developed. Due to its complex and irregular shape, the glenoid part of a left embalmed scapula bone was chosen as working material. CT data were retrieved using a standard clinical CT scanner (Siemens Somatom Plus 2, Siemens AG, Germany). This was done to produce a method that could later be utilized to generate a patient-specific FE model. Different methods of converting Hounsfield unit (HU) values to apparent densities and subsequently to Young's moduli were tested. All the models obtained were loaded using three-dimensional loading conditions taken from literature, corresponding to an arm abduction of 90 degrees. Additional models with different amounts of elements were generated to verify convergence. Direct comparison between the models showed that the best method to convert HU values directly to apparent densities was to use different equations for cancellous and cortical bone. In this study, a reliable method of determining both geometrical data and bone properties from patient CT scans for the semi-automated generation of an FE model is presented. PMID- 11382080 TI - A new medical microrobot for minimal invasive surgery. AB - A new medical microrobot to drive endoscopes into endocoeles is presented. The structure of the microrobot is very simple; it consists of a right spirally grooved micromotor, a left spirally grooved cylinder and a flexible coupling. When the micromotor rotates, a hydrodynamic mucus film is formed because highly viscous mucus exists in endocoeles. The mucus film can prevent direct contact between the microrobot and the endocoele, and the injury to organic tissues may be avoided. The locomotion speed and the hydrodynamic mucus film thickness formed when the microrobot drives endoscopes into endocoeles have been calculated according to hydrodynamic lubrication theory. The results indicate that the microrobot can be suspended to drive endoscopes quickly into endocoeles. This has been confirmed by experiments. PMID- 11382081 TI - Modelling femoral curvature in the sagittal plane: a cadaveric study. AB - This study examined the possibility of representing the mid third of the human femur with two straight sections. This portion of the femur visually has a distinct curvature, which can potentially present problems when considering implant stem designs to be introduced in this region. Sixteen femora were sectioned at 10 mm intervals along the femoral shaft in the mid third region (35 65 per cent of femoral length). Photographic records were obtained of each section against a consistent axis system to which all coordinates were referenced. The position of the centre of the medullary canal cross-sectional area along the femur, in relation to fixed orthogonal planes, has been analysed; the outer anterior cortex was also modelled. The results showed that the medullary centre of area plots and the anterior cortex coordinates are suitably modelled as two straight lines. For each bone it was possible to define the intersection point between the two straight sections (point of angulation), and the subtended angle between these sections (angle of incidence). The average point of angulation for the medullary plots occurred at 57 per cent along the femur, while the mean angle of incidence was 6.5 degrees. The anterior surface had an average point of angulation at 58 per cent along the femur with the mean angle of incidence being 22.2 degrees. The centre-line of the medulla was also found to be almost parallel to the outer anterior surface for sections distal to the point of angulation. It is proposed therefore, that this difference in angulation is the result of medullary expansion/cortical thinning towards the proximal extremity of the femur, causing the straight-line model of the medulla to angulate less than the outer anterior cortex. PMID- 11382082 TI - The development of a modular humeral prosthesis locking mechanism. AB - A new modular humeral prosthesis has been developed, incorporating a novel locking mechanism to prevent head-stem disassembly. A series of test protocols was designed to evaluate the integrity of the new locking mechanism and a dedicated test rig was manufactured. Preliminary test results were disappointing and the locking mechanism failed at lower applied loads than predicted. Microscopy studies were conducted to establish the cause of this poor performance and with the aid of statistical analysis, these studies assisted in the development process. Development of the design was directed at the control of the position and angle of contact between bearing surfaces of the locking mechanism. The optimized design was tested under the maximum anticipated loading conditions and performed satisfactorily. PMID- 11382083 TI - Unsteady viscous flow model on moving the domain through a stenotic artery. AB - An unsteady Navier-Stokes (N-S) solver based on the method of operator splitting and artificial compressibility has been studied for the moving boundary problem to simulate blood flow through a compliant vessel. Galerkin finite element analysis is used to discretize the governing equations. The model has been applied to a time-varying computational domain (two-dimensional tube) as a test case for validation. Consideration has been given to retaining the space conservation property. The same code is then applied to a hypothetical critical high-pressure gradient over a short length of blood vessel based on the spring and dashpot model. The governing equation for the blood vessel is based on two dimensional dynamic thin-shell theory that takes into account the curvature of the stenotic portion of the vessel. Progressing the solution towards steady state is considered, as the main objective is to show the viability of the current technique for fluid/structure interactions. Preliminary results of the wall velocity and displacement based on steady state prediction agree well with data in the literature. Results, such as the streamlines, wall pressures and wall shear stress depict the possible progression of arterial disease. PMID- 11382084 TI - The effect on the fatigue strength of bone cement of adding sodium fluoride. AB - New bone cements that include several additives are currently being investigated and tested. One such additive is sodium fluoride (NaF), which promotes bone formation, facilitating implant integration and success. The influence of NaF on the fatigue performance of the cement as used in biomedical applications was tested in this paper. In fact fatigue failure of the cement mantle is a major factor limiting the longevity of a cemented implant. An experimental bone cement with added NaF (12 wt%) was investigated. The fatigue strength of the novel bone cement was evaluated in comparison with the cement without additives; fatigue tests were conducted according to current standards. The load levels were arranged based on a validated, statistically based optimization algorithm. The curve of stress against number of load cycles and the endurance limit were obtained and compared for both formulations. The results showed that the addition of NaF (12 wt%) to polymethylmethacrylate (PMMA) bone cement does not affect the fatigue resistance of the material. Sodium fluoride can safely be added to the bone cement without altering the fatigue performance of the PMMA bone cement. PMID- 11382085 TI - Comparative two- and three-dimensional finite element modelling techniques for tibial fractures. AB - Often the choice of a two-dimensional modelling approach over a three-dimensional approach is made on the basis of available resources, and not on task appropriateness. In the case of simulating the mechanical behaviour of irregular anatomical structures in biomedical engineering, the authenticity of two dimensional model behaviour and the interpretation of model solutions is of particular concern since little comparable two-dimensional and three-dimensional data have been published. As part of a research programme, a comparison was made between two-dimensional and three-dimensional finite element models (FEMs) that examine the stress-strain environment of a clinical bone fracture and callus. In comparison with the three-dimensional model, the two-dimensional model substantially underestimated peak compressive principal stresses in the callus tissue and peak equivalent strains. This was a consequence of geometrical and structural asymmetry in a plane perpendicular to the two-dimensional model. However, the two-dimensional model predicted similar patterns of stress and strain distribution to the corresponding mid-longitudinal plane of the three dimensional model, and underestimates of peak stress and strain were much reduced. This confirmed that despite the irregular geometry and structure of the subject, the two-dimensional model provided a valid mechanical simulation in the plane of the fracture that it represented. PMID- 11382086 TI - The influence of phospholipid concentration in protein-containing lubricants on the wear of ultra-high molecular weight polyethylene in artificial hip joints. AB - There is considerable interest in the wear of polyethylene and the resulting wear debris-induced osteolysis in artificial hip joints. Proteins play an important role as boundary lubricants in vivo in the pseudosynovial fluid, and these are reproduced in in vitro tests through the use of bovine serum. Little is known, however, about the effect of phospholipid concentrations within proteinaceous solutions on the wear of ultra-high molecular weight polyethylene (UHMWPE). The effects of protein-containing lubricants with 0.05, 0.5 and 5 per cent (w/v) phosphatidyl choline concentrations on the wear of ultra-high molecular weight polyethylene (UHMWPE) were compared with 25 per cent (v/v) bovine serum which had 0.01 per cent (w/v) lipid; the effects were compared in a hip joint simulator with smooth (n = 4) and scratched (n = 3) femoral heads. The control bovine serum lubricant produced UHWMPE wear of 55 and 115 mm3/10(6) cycles on the smooth and rough heads respectively. The increased phospholipid concentration significantly reduced the wear rate. At the higher concentration (5% w/v phosphatidyl choline) the average wear was reduced to less than 2 mm3/10(6) cycles. Even with the relatively low concentrations of 0.05% w/v phosphatidyl choline the wear was reduced by at least threefold compared with the bovine serum tests for both the smooth and rough femoral heads. There may be considerable differences in the phospholipid concentrations in patients' synovial fluid and this is highly likely to produce considerable variation in wear rates. In vitro, differences in the phospholipid concentration of lubricants may also cause variation in wear rates between different simulator tests. PMID- 11382087 TI - Alice Catherine Evans (1881-1975). PMID- 11382088 TI - Malaria kills one child every 30 seconds. PMID- 11382089 TI - A short history of occupational health. PMID- 11382090 TI - Epidemiologic highlights of the past with a look towards the future. AB - Since the first meeting of the American Public Health Association in 1873, epidemiology and epidemiologists have been central to the activities of the organization. At that meeting, the most prominent American physician of the time, Austin Flint, presented a classic paper entitled "Relations of Water to the Propagation of Fever." In that remarkable paper, Flint reinterpreted observations on the North Boston, N.Y. typhoid fever epidemic made 30 years earlier in which he had correctly concluded that the epidemic had been propagated by contagion, but incorrectly concluded that transmission was not the result of contaminated drinking water. During the last quarter of the 19th century and the first quarter of the 20th, Annual Meetings of the APHA were frequently the locus for reports of epidemic investigations and the Journal of the Association was the vehicle for their publication. When the Association was reorganized to include discipline oriented sections, the Epidemiology Section was among the first to be established. At the time, 1929, the Section was the only meeting place (outside the small and exclusive American Epidemiological Society) for epidemiologists to exchange ideas and information. The Section was also a place where public health policies were hotly debated. After World War II, academic departments of epidemiology increasingly focused the field on methodological issues and etiological investigations. Furthermore, new organizations and a plethora of epidemiological journals flooded the field. Thus, the Epidemiology Section has remained a major center for translating epidemiological knowledge into policy and is likely to intensify that function in the future. PMID- 11382091 TI - Eliminating smoking in bars, restaurants, and gaming clubs in California: BREATH, the California Smoke-Free Bar Program. AB - On January 1, 1998, California became the first state in the country to prohibit indoor smoking in nightspots, bars, bar/restaurant combinations, bingo parlors, cardrooms, and casinos on a statewide basis. This paper describes the activities which achieved this result, based on a program with three main goals: Ease the transition for business owners; Activate public support for the law; Defeat tobacco industry efforts to undermine the law. Important lessons learned from this campaign are presented. PMID- 11382092 TI - Estimating the costs of caring for people with Alzheimer disease in California: 2000-2040. AB - The costs of caring for people with Alzheimer disease (AD) in California are estimated using data from a study of the costs of caring for community-resident and institutionalized people with AD, combined will prevalence and population projections. Costs for community-resident patients will increase 83 percent in the period 2000 ($23.4 billion) to 2020 ($42.8 billion), and will grow an additional 59 percent from 2020 to 2040 ($68.1 billion). Costs for AD patients in institutions will increase 84 percent from 2000 ($2.5 billion) to 2020 ($4.6 billion), and will grow an additional 61 percent from 2020 to 2040 ($7.4 billion), assuming the supply of nursing home beds meets projected demand. Total costs of caring for AD patients will nearly triple between 2000 and 2040. The rapid aging of the U.S. population makes more aggressive societal action necessary if the personal and societal burden of Alzheimer's disease is to be reduced in the future. PMID- 11382093 TI - Carlos Juan Finlay (1833-1915). PMID- 11382094 TI - Evidence-based medicine. A critical view from the clinician. PMID- 11382095 TI - [Gene therapy: its contributions to liver and kidney diseases]. PMID- 11382096 TI - [Mechanisms responsible for peripheral vasodilation in liver cirrhosis]. PMID- 11382097 TI - [Hepatorenal syndrome]. PMID- 11382098 TI - [Cell activation and apoptosis as markers of hemodialysis-induced inflammation]. PMID- 11382099 TI - [Physiopathologic bases and data of the general population]. PMID- 11382100 TI - [Polymorphisms of the vitamin D receptor (VDR) gene and parathyroid function]. PMID- 11382101 TI - [Polymorphism of the vitamin D receptor and bone disease after renal transplantation]. PMID- 11382102 TI - [Vascular oxidative stress and endothelial dysfunction]. PMID- 11382103 TI - [Polymorphisms of the ACE gene and cardiovascular disease]. PMID- 11382104 TI - [What is the optimal treatment duration of group A streptococcal pharyngitis?]. PMID- 11382105 TI - [Primary carcinoma of the ear canal. Clinical aspects, intratemporal growth behavior and surgical strategy]. AB - BACKGROUND AND OBJECTIVE: The symptoms and microscopic findings in carcinoma of the external auditory meatus are often unspecific and misinterpreted as recurrent otitis externa. A definite diagnosis is therefore often delayed, frequently resulting in advanced tumor stages with unfavorable prognosis at the time of diagnosis. PATIENTS/METHODS: In a retrospective analysis of 11 of our own patients and four cases from the Wittmack collection, the clinical data and course of this disease were examined with particular focus on the intratemporal bone growth pattern and possible surgical strategies. RESULTS: Histopathologically, all cases consisted of squamous cell carcinoma of variable differentiation, with predomination of higher tumor stages (T2-T4: 93.3%) and little initial lymph node metastases (N+: 14.3%; M+: 0%). Osseous destruction was present (almost obligatory) in 93.3% (determined histopathologically and radiologically with CT). Follow-up (to a maximum of 6 years) after surgical resection and postoperative irradiation shows local recurrence at an average of 10 months in 40% of the cases, with a metastasis rate of 44.4%. Mean overall survival is 29.7 months, and 45.5% of the patients experience an average tumor free period of 46.4 months. The most important prognostic factor in this context is the initial tumor stage. CONCLUSION: In regard to prognosis of external auditory meatus carcinoma, early diagnosis is of vital importance. Especially older patients (50-80 years) who previously have had no history of ear disease should undergo early computer tomographical examination and biopsy in cases of recurrent external otitis. PMID- 11382106 TI - [Clinical aspects and therapy of extratemporal facial neurinoma]. AB - BACKGROUND AND OBJECTIVE: Extratemporal facial nerve neurinomas are rare. In the present study, the epidemiology, clinical presentation and in particular surgical treatment of this entity was analysed under special focus on facial nerve preservation. PATIENTS/METHODS: 22 patients with an extratemporal facial nerve neurinoma of the parotid gland, treated at the University-ENT-Clinic Hamburg Eppendorf during 1977-1997 were analysed retrospectively. RESULTS: All patients presented with a unilateral parotid mass, on ultrasound examination regularly an encapsulated intraparotideal tumor. 16 patients (72.7%) had regular facial nerve functioning. 6 patients (27.3%) with partial or complete peripheral facial nerve paralysis were treated with partial or radical parotidectomy because of malignancy possibility. In the group of patients with intact facial nerve, latero facial (superficial) parotidectomy with segmental nerve resection was undertaken in 9 cases (56.3%) because nerve-continuity was not siezable within the tumor area. In 7 cases (43.7%), tumor-enucleation was feasable with complete nerve preservation. Intraoperative frozen section in a total of 4 (18.2%) patients always showed the correct histopathological diagnosis. No tumor-recurrence was observed in the 15 cases (68.6%) which underwent follow-up. CONCLUSIONS: In case of extratemporal neurinoma of the parotid gland, preparation of the facial nerve during surgery is often impossible, leading to segmental nerve resections. This decision, however, is made without proof for malignancy. In recognition of the biological behaviour of this entity the authors recommend that after tumor exploration a biopsy should be taken from the periphery of the tumor to exclude malignancy. After diagnosis of a neurinoma a nerve preserving (wait-and-see) approach appears justified. PMID- 11382107 TI - [Sleep-associated respiratory disorders in childhood]. AB - Sleep-related breathing disorders in children are common. Nearly 10% of preschool aged children snore, and 1% of 4- to 5-year-old children present with an obstructive sleep apnea syndrome (OSAS), mostly due to an adenotonsillar hyperplasia. OSAS in children differs markedly from adults concerning etiology, clinical symptoms, polysomnographic findings, and course of the disease. Therefore, results of adult sleep medicine cannot easily be applied to children. The disease may result in pulmonary or systemic hypertension, failure to thrive, and neurocognitive misbehavior. Up to now, there is no consensus concerning diagnosis and therapy. In this article, we summarize and discuss what is known so far about sleep-related breathing disorders in children, focussing on the OSAS as the most important diagnosis for the ENT specialist. PMID- 11382108 TI - [Incidence of salivary fistulas in relation to timing of oral nutrition after laryngectomy]. AB - BACKGROUND AND OBJECTIVE: A pharyngocutaneous fistula is the most common complication after total laryngectomy. In Germany, a traditional recommendation is to use a nasogastric tube for feeding for 10-14 days postoperatively because many surgeons believe that oral feeding after surgery contributes to fistula development. However, there is no international agreement about when to begin oral feeding after total laryngectomy. Some authors begin oral feeding between the 1st and 4th postoperative day without any nasogastric tube, while others using a nasogastric tube delay oral feedings until 7-14 days after surgery. The aim of the present study was to investigate the relationship between the timing of oral feeding and the development of fistulas after total laryngectomy. PATIENTS/METHODS: In a prospective trial with 42 consecutive patients who underwent laryngectomy, oral feeding was started on different postoperative days between the 1st and the 10th. Most patients were selected randomly for the different postoperative days. Furthermore, other potential risk factors predisposing to fistula formation were analyzed retrospectively. RESULTS: Five fistulas occurred in the total group (12%). Early postoperative oral feeding does not increase the incidence of fistulas. The fistula rate was only 9% in patients fed orally in the 1st postoperative week. The analysis of further risk factors for fistula formation showed only a significant correlation between type of resection and fistula occurrence (extended laryngectomy with partial pharyngectomy vs standard laryngectomy; p = 0.018). CONCLUSIONS: Our results indicate that early oral feeding in the 1st postoperative week does not influence fistula formation after laryngectomy. PMID- 11382109 TI - [Squamous epithelial carcinomas of the external ear]. AB - BACKGROUND AND OBJECTIVE: Squamous cell carcinoma of the pinna seems to be associated with a worse prognosis as compared to other locations. PATIENTS/METHODS: We studied 88 patients treated between 1975 and 1990 for a squamous cell carcinoma of the pinna. RESULTS: Lymph node metastases were present in eight cases (9%) prior to treatment. Treatment was intended to be curative in 83 patients (94%). Tumor therapy was operative in all cases. Radiotherapy was instituted postoperatively in three patients; five patients (5.7%) died due to the tumor. Of 83 curatively treated patients, only 2 died of tumor progression. The survival rate was 98% after 2 years and 95% after 5 years. The recurrence rate was 7% after 1 year, 13% after 2 years, and 18% after 5 years. The outcome with regard to local tumor control and survival was significantly poorer when neck metastases were present. CONCLUSIONS: We recommend tumor excision with wide margins (5-10 mm) as first-line treatment. Neck dissection with parotidectomy is indicated when suspicious lymph nodes are detected by ultrasound sonography, the tumor diameter is > 4 cm, cartilaginous invasion is present, and vertical tumor thickness is > 5 mm. PMID- 11382110 TI - [Value of acoustic speech analysis for prognostic assessment of stuttering in children. Partial results of a prospective longitudinal study]. AB - PRESENT STATE OF KNOWLEDGE: There are currently no known acoustic parameters by which stuttering children can be appraised which will predict their subsequent speech fluency. AIM: To explain the significance of factors which perpetuate stuttering by using computer-based speech analysis of fluent speech for a 1 1/2 year period and to relate acoustic analysis with clinical measurements of stuttering. Special attention was given to motor-oral and/or linguistic deficits. METHOD AND RESULTS: A prospective study of 58 pre-school children who stutter. Correlations were sought between acoustic variables in the severity and course of the stuttering with the influence of motor-oral and linguistic disturbances. 19 age-matched, normal-speaking children served as controls. A subdivision of the study group into different subgroups with particular motor-oral and/or linguistic problems showed that children whose stuttering coincides with a delayed speech development have a distinctly better prognosis for early remission. In most of these children the stuttering remitted to such a degree as the deficits causing the stuttering could be reappraised, which means simultaneous improvement of the linguistic competence. CONCLUSION: It was apparent that remission rate was much higher in those children who showed linguistic disturbances at the same time with stuttering. Within the stuttering group, subgroups showed a few correlations in several acoustic parameters, but these could not, as yet, be shown to give any prognostic markers in the routine diagnosis of children who stutter. If a child shows any danger-signs of acquiring stuttering on a more permanent basis, a careful diagnosis is necessary in order to find the individually underlying factors before any therapeutical procedure. PMID- 11382111 TI - [Metastasis of a spinal myxopapillary ependymoma to the inner auditory canal]. AB - Ependymomas are usually benign tumors of the central nervous system that derive from the ependymal cells lining liquor-filled spaces. Myxopapillary ependymomas are a variant occurring almost exclusively in the region of the cauda equina. Histologically, these tumors are benign (WHO grade I). The following case describes the first reported spread of a spinal myxopapillary ependymoma into the internal auditory canal. The symptoms and diagnostic results are similar to other tumors of this location. There are no typical findings in the diagnostic imaging (CT, MRI). The extended middle cranial fossa approach was chosen for surgical treatment. Adjuvant radiotherapy is indicated in cases of incomplete resection or recurrence. PMID- 11382112 TI - [Hyperacusis after tympanoplasty]. AB - A case of hyperacusis after tympanoplasty is presented. A 62-year-old woman experienced unilateral conductive hearing loss for about 60 years after antrotomy in infancy. Neurological or otoneurological symptoms were not evident. Tympanotomy showed a missing incus with discontinuity of the chain. Reconstruction was performed by interposition of a partial ossicular replacement prosthesis (PORP). The postoperative audiogram revealed minimal perceptive hearing loss in the high-frequency field in the operated ear. Several weeks after middle ear surgery, the patient complained of hyperacusis on the operated side, which persisted for more than 2 years. To alleviate the unpleasant sensations, an ear plug was used by the patient. In spite of the asymmetric hearing loss, no late onset auditory deprivation could be diagnosed. The pathophysiological causes of the hyperacusis have not yet been clarified. Possible reasons are discussed. PMID- 11382113 TI - [Psychogenic vertigo in ENT practice]. PMID- 11382114 TI - [Unilateral exophthalmos of uncertain origin. Orbital emphysema in medial "blow out fracture"]]. PMID- 11382115 TI - [Penetrating irrigation of the maxillary sinus]. PMID- 11382116 TI - [Tuberculous and nontuberculous mycobacterial diseases of the cervical lymph nodes]. PMID- 11382117 TI - [Segmental type 2 manifestation of autosome dominant skin diseases. Development of a new formal genetic concept]. AB - The prevailing theory says that mosaic forms of autosomal dominant skin diseases originate from postzygotic new mutations. This theory is no longer generally valid. According to a new rule of dichotomy, we can distinguish two types of segmental manifestations. The type 1 reflects heterozygosity for a postzygotic new mutation, whereas the type 2 results from loss of the corresponding wildtype allele occurring in a heterozygous embryo and reflects either homozygosity or hemizygosity for the underlying mutation, giving rise to rather pronounced segmental lesions that are superimposed on the ordinary nonsegmental phenotype. Autosomal dominant skin diseases exemplifying the concept of type 2 segmental manifestation so far include neurofibromatosis 1, tuberous sclerosis, cutaneous leiomyomatosis, glomangiomatosis, Buschke-Ollendorff syndrome, multiple syringomas, multiple trichoepitheliomas, multiple basaloid follicular hamartomas, multiple nevoid basal cell carcinomas, Darier disease, Hailey-Hailey disease, epidermolytic hyperkeratosis of Brocq, KID syndrome, disseminated superficial actinic porokeratosis and autosomal dominant dyskeratosis congenita. A strikingly high frequency of type 2 segmental involvement has been documented in cutaneous leiomyomatosis, glomangiomatosis and disseminated superficial actinic porokeratosis. It should be noted that there is so far no molecular proof for the proposed rule of dichotomy that has been developed from clinical dermatology. According to present knowledge, however, it is very likely that molecular analysis will confirm the described concept that can explain some so far enigmatic features as observed in autosomal dominant genodermatoses. PMID- 11382118 TI - [Rare and newly described histological variants of cutaneous squamous epithelial carcinoma. Classification by histopathology, cytomorphology and malignant potential]. AB - The histological spectrum of squamous cell carcinoma (SCC) of the skin is presented, including recently mentioned and rarely appearing tumor subtypes. Beside the most frequently occurring common SCC with variable degree of differentiation, the following tumor forms can be identified microscopically: Bowen-SCC, acantholytic SCC, clear cell SCC, spindle cell SCC, keratoacanthoma like SCC, desmoplastic SCC, neurotrophic SCC, verrucous SCC, adenosquamous carcinoma and basosquamous cell carcinoma. The classification of lymphoepithelioma-like carcinoma as a cutaneous SCC variant is disputed, because epidermal connections are regularly absent and evidence of glandular and follicular differentiation suggest an adnexal origin. Beside the description of distinctive histological and cytomorphological features of each SCC subtype, important differences in malignant behavior and clinical course are emphasized. PMID- 11382119 TI - [Disseminated melanoma cells in blood and bone marrow. Significance and detection by potential tumor markers]. AB - As the majority of primary malignant melanomas can be cured by surgical excision, the prognosis of melanomas is dependent on whether tumor cells have disseminated orare capable of doing so at the time of surgery. A prospective and valid detection of this minimal residual disease is not currently possible. The most important known so-called markers of melanoma disease, tyrosinase, S100 and MIA, all are more likely to be present in patients with more advanced disease. A valid prognostic effect has only been shown for S100 in patients with already identified metastatic disease. Further prospective studies are required to determine the potential gain of information by routine determination of these markers in melanoma patients. PMID- 11382120 TI - [Guidelines for quality assurance in ultrasound diagnosis in dermatology. Guideline of the Ultrasound in Dermatology Subgroup of the Physical procedures in Dermatology Subcommittee January 2000]. PMID- 11382121 TI - [Botulinophilia. The new life style venenophilia]. AB - BACKGROUND AND OBJECTIVE: Botulinum toxin is effective in the treatment of hyperhidrosis and the demand for therapy is increasing. Simultaneously we have observed an increase in patients with body dysmorphic disorders who also want botulinum toxin therapy. This botulinophilie is a new variant of venenophilie. We investigated the prevalence of this new diagnosis in our patient population. PATIENTS/METHODS: In the first quarter of 2000 we studied the biopsychosocial features of 13 patients with hyperhidrosis. RESULTS: In 23.1% of our cases we were able to confirm a botulinophilie with body dysmorphic disorder and a normal Minor sweat test. CONCLUSIONS: Botulinophilie is not an indication for botulinum toxin therapy but for psychotherapy. PMID- 11382122 TI - [What remains of the skin after 2000 years in a bog?]. AB - BACKGROUND AND OBJECTIVE: Mummies have an important place in the study of archaeology and paleopathology because they are so well preserved. For the first time skin samples of six 2300-1600 year old bog bodies from North Germany were examined by histology, transmission electron microscopy and immunohistology. METHODS: For histology the samples were stained with H&E and van Gieson elastic stain. Fixation and embedding in epoxy followed for the transmission electron microscopy. Specific antibodies directed to type IV collagen and S-100 were used. RESULTS: Histologically it was possible to observe collagen bundles in the dermis, with a density similar to recently stained samples. Epidermis was not preserved. The electron microscopy showed collagen fibrils with a diameter of 45 110 nm and the characteristic axial periodicity. Throughout the dermis, a number of spores of bacteria with a diameter of 0.83 +/- 0.051 micron and an electron dense core were found. No activity against the used antibodies could be detected. CONCLUSIONS: Histology and electron microscopy demonstrate the excellent conservation of the dermal collagen in the bog. In contrast to ice mummies like "Otzi" and mummies from Egypt, no cellular elements could be found in the skin of bog bodies. PMID- 11382123 TI - [Photochemotherapy of cutaneous AIDS-associated Kaposi sarcoma with indocyanine green and laser light]. AB - BACKGROUND AND OBJECTIVE: Indocyanine green (ICG) is a clinically approved dye for diagnostic purposes, which has an absorption peak in the near infrared and remains intravascular due to a high plasma protein binding. Its therapeutic potential in combination with a diode laser was studied for well vascularized cutaneous tumors. PATIENTS/METHODS: Six male patients (mean age 49.2 years) with AIDS-related Kaposi sarcomas (n = 30) received ICG (2 x 2 mg/kg i.v.) followed directly by irradiation with a diode laser (lambda = 805 nm, 100 J/cm2, 3 W/cm2). RESULTS: All macular and plaque-type lesions (n = 27) showed primarily blister- and crust formation and healed within 14 days. Only one out of the 3 nodular lesions treated showed complete remission. The only side effect recognized was a mild burning sensation during irradiation. Nineteen lesions resolved completely leaving a slight atrophic scar, in three lesions a transient postinflammatory hyperpigmentation occurred. Within the follow-up period of 2 years no recurrence was detected. CONCLUSIONS: The ICG-mediated photochemotherapy is an effective palliative therapeutic modality with a low rate of side effects in the treatment of macular or plaque-type cutaneous Kaposi sarcomas. PMID- 11382124 TI - [Case report on therapy with granulocyte stimulating factor in diabetic foot]. AB - Several pathogenetic factors such as peripheral neuropathy, vasculopathy and infection are responsible for the development of diabetic foot ulcerations. An important factor contributing to the high infection risk in diabetic patients is a defect in neutrophil granulocytes. Deficiencies in neutrophil chemotaxis, phagocytosis and respiratory burst activity with the decrease of the super- and peroxids are known to be associated with diabetes. Granulocyte-colony stimulating factor (G-CSF) increases the release of neutrophils from the bone marrow and improves neutrophil function. A 78-year old patient with non-insulin-dependent diabetes presented with ulcerations of both big toes and a malum perforans on the right sole. He also had generalized arteriosclerosis as well as a polyneuropathy with a dry foot and typical foot deformation as well as decreased in sensitivity. Intensive local care for 35 days led to no improvement of the ulcerations. Then G CSF (Neupogen) was administered in a total dose of 165 million IU over 11 days; the daily dose varied between 15-30 million IU depending on the absolute leucocyte count. In addition 500 mg of oral ciprofloxacin (Ciproxin) was given b.i.d. This treatment led to a significant improvement of the lesions. Within 11 days cost analysis suggests G-CSF may be a cost-effective addition to antimicrobial therapy in diabetic foot infection. PMID- 11382125 TI - [Postherpetic erythema exsudativum multiforme with peri-nevus collection of annular outbreaks]. AB - A 10 year old boy with postherpetic erythema multiforme developed an unusual arrangement of target lesions around all preexisting melanocytic nevi on his trunk. This is the first description of a perineval erythema multiforme. PMID- 11382126 TI - [Bilateral asymmetric herpes zoster in adolescence]. AB - Zoster is a frequent disease of adulthood with a distinct age-dependent increase after 60. In contrast during childhood or adolescence zoster only rarely occurs. Certain risk factors such as hematologic malignancies are associated with early appearance. The typical clinical manifestation is unilateral, equally involving thoracic dermatomes. A 16-year-old patient presented with zoster in bilateral asymmetrical distribution, with trigeminal and thoracic dermatomes simultaneously affected. Despite the clinical findings and the unusual localization, there was no history, clinical nor laboratory signs of an immune suppression or any other underlying disease. Careful follow-up examinations are necessary in order to recognize systemic, especially hematologic, malignancies. PMID- 11382127 TI - [Systematic sebaceous nevus with multiple secondary tumors as the chief symptom of Schimmelpenning-Feuerstein-Mims syndrome]. AB - A women presented with a systematized sebaceous nevus of the head and neck. She had developed multiple basal cell carcinomas and sebaceomas in the nevus over the past 3 years after a period of about 55 years without any changes. Detailed medical history and physical examination revealed a constellation of findings suggesting a syndrome. Besides ptosis and a congenital, vascular corneal clouding at the right, there was hypoplasia of the right maxillary sinus, a right labyrinthine deafness and recurrent inguinal herniae on the right. The Schimmelpenning-Feuerstein-Mims-Syndrome was diagnosed. PMID- 11382128 TI - [Isolated positive antistreptolysin titer in chronic urticaria or psoriasis vulgaris]. PMID- 11382129 TI - [Ernst Schweninger, personal physician of the German chancellor and director of the Charite Dermatology Clinic 1884-1902]. AB - Ernst Schweninger was at the age of 33 years already world renowned for being Bismarck's personal physician. In 1884, his grateful patient made him chief of the Department of Dermatology in Berlin and Professor of Dermatology. The subsequent years were marked by political struggles within the university, but he still produced a number of scientific publications, and is still remember for his first description of anetoderma type Schweninger-Buzzi. Dermatology and his activities at the Berlin clinic from whose direction he resigned in 1902 accounted for only one period in his life. His main interest was the physiologic dietetic therapeutic method which he applied also to dermatology. Later on, he dissociated himself from scientific medicine and became the founder of a school for natural healing methods. The present report is designed to keep alive the memories of this politically active physician. PMID- 11382130 TI - [Comment on the contribution by H. Breuninger, C. Garbe and G. Rassner: Shave excision of melanocytic nevi of the skin--indications, technique, results]. PMID- 11382131 TI - [Swelling and erythema of the right leg in a 62-year-old patient. Wells syndrome]. PMID- 11382132 TI - [Infections with varicella zoster virus]. PMID- 11382133 TI - [Living and letting die--new mechanisms of apoptosis resistance in melanoma cells]]. PMID- 11382134 TI - The origin of life. I: When and where did it begin? AB - For decades most scientists assumed that life emerged billions of years ago in a "primordial soup" somewhere on the Earth's surface. Evidence is mounting, however, that life may have begun deep beneath the surface, perhaps near a volcanic ocean vent or even inside the hot crust itself. Since there are hints that life's history on Earth extends back through the phase of massive cosmic bombardment, it may be that life started on Mars and came here later, perhaps inside rocks ejected from the Red Planet by large impacts. The traffic of intact rocks between Mars and Earth is now an established fact, and experiments confirm that microbes could survive the rigours of the journey through space if cocooned within such material. Unfortunately, this planetary cross-contamination compromises astrobiologists' hope of finding a second genesis in the solar system. PMID- 11382135 TI - The origin of life. II: How did it begin? AB - The problem of how a mixture of chemicals can spontaneously transform themselves into even a simple living organism remains one of the great outstanding challenges to science. Various primordial soup theories have been proposed in which chemical self-organization brings about the required level of complexity. Major conceptual obstacles remain, however, such as the emergence of the genetic code, and the "chicken-and-egg" problem concerning which came first: nucleic acids or proteins. Currently fashionable is the so-called RNA world theory, which casts RNA in the role of both chicken and egg. Other theories assume that protein chemistry and even clay crystal life came before nucleic acids. To be fully successful, a theory of biogenesis has to explain not merely the emergence of molecular replication and chemical complexity, but the crucial information content and information processing capabilities of the living cell. PMID- 11382136 TI - Relics: penguin population programs. AB - What has been responsible for the increase in Chinstrap penguin populations during the past 40 years in maritime Antarctica? One view ascribes it to an increase in availability of their prey brought on by the decrease in baleen whale stocks. The contrary opinion, attributes it to environmental warming. This causes a gradual decrease in the frequency of cold years with extensive winter sea ice cover. A number of penguin monitoring programs are in progress and are expected to provide some answers to these questions. Unfortunately, it is not easy to distinguish natural variability from anthropogenic change since penguins are easily accessible predators of krill and the feeding range of the penguins has almost overlapped with the krill fishery in time and space in the last four decades. Therefore it is important to reconstruct the change of ancient penguin abundance and distribution in the absence of human activity. Many efforts have focused on surveying the abandoned penguin rookeries, but this method has not been able to give a continuous historical record of penguin populations. In several recent studies, ancient penguin excreta was scooped from the penguin relics in the sediments of the lake on penguin rookery, Ardley Island, maritime Antarctica. In these studies, penguin droppings or guano soil deposited in the lake and changes in sediment geochemistry have been used to calculate penguin population changes based upon the geochemical composition of the sediment core. The results suggest that climate change has a significant impact on penguin populations. PMID- 11382137 TI - Use of EPR spectroscopy to study macromolecular structure and function. AB - Electron paramagnetic resonance (EPR) spectroscopy is now part of the armory available to probe the structural aspects of proteins, nucleic acids and protein nucleic acid complexes. Since the mobility of a spin label covalently attached to a macromolecule is influenced by its microenvironment, analysis of the EPR spectra of site-specifically incorporated spin labels (probes) provides a powerful tool for investigating structure-function correlates in biological macromolecules. This technique has become readily amenable to address various problems in biology in large measure due to the advent of techniques like site directed mutagenesis, which enables site-specific substitution of cysteine residues in proteins, and the commercial availability of thiol-specific spin labeling reagents (Figure 1). In addition to the underlying principle and the experimental strategy, several recent applications are discussed in this review. PMID- 11382138 TI - Safety aspect concerning radiolytic gas generation in reactors. AB - In water cooled and water moderated reactors (H2O in boiling water reactors/pressurised water reactors, D2O in pressurised heavy water reactors) during normal operation, radiolysis is a source of production of hydrogen/deuterium and oxygen. During the progress of a nuclear accident, while there are other important sources of hydrogen/deuterium, the oxygen availability can occur only through radiolysis or direct contact with air. In air saturated with water vapour at room temperature and pressure when H2/D2 concentration exceeds 4 vol % (a conservative estimate), a combustible mixture with oxygen can be formed. It is proposed to examine the basic principles of water radiolysis as far as they pertain to generation of H2/D2 and O2 and try to apply these concepts to reactors both under operating conditions and in accident situations. It is concluded that the possibility of an accident taking place through radiolysis is highly unlikely. PMID- 11382139 TI - [Tumor markers]. AB - Tumour markers are substances developed in or induced by tumour cells and secreted into body fluids in which they can be quantified by non-invasive analyses. The malignant transformation of cells leads to increased concentrations of tumour markers and thus they can indicate malignant diseases. It appears, however, that other proliferative processes, i.e. inflammatory and benign transformations are also able to induce the rise of tumour marker levels. Due to their low sensitivity and specificity, tumour markers--except for PSA--are not useful in diagnosis and screening. Though disseminated malignant disorders are associated with high tumour marker levels, a correlation between their concentration and the tumour volume is not clearly approved. The use of tumour markers seems established for the follow-up after curative surgery and for the treatment and monitoring of palliative therapy. PMID- 11382140 TI - Bone mineral density in multiple myeloma patients after intravenous clodronate therapy. AB - Bisphosphonates inhibit osteoclastic bone destruction that may be stimulated by myeloma cells. By this way, bisphosphonates carry the potential to lower the number of new pathological fractures, of hypercalcaemic events, and of intensity of bone pain as was published earlier. Clodronate has been administered orally in most clinical studies so far despite of its poor bioavailability from the gastrointestinal tract. The aim of our study was to evaluate bone mineral density (BMD) changes in 34 newly diagnosed multiple myeloma patients regularly treated by 900 mg of clodronate in slow intravenous infusions on an outpatient basis in two-week intervals for at least 24 months. BMD was evaluated by CT scanning every six months. Initial values of trabecular BMD were only about 60 per cent of age- and sex-adjusted healthy population values. Clodronate therapy seemed to preserve BMD within the period of two or three years. This effect was seen not only in patients responding to chemotherapy but even in the small subgroup of patients with persisting active disease. We conclude that the long-term intravenous clodronate therapy may contribute to the preservation of BMD and thus calcium hydroxyapatite in bones in multiple myeloma patients concomitantly treated by chemotherapy. The administration of clodronate should start early in the course of disease, before BMD has been markedly reduced. PMID- 11382141 TI - The limits of surgical treatment of non-small cell lung cancer. AB - Between January 1, 1991 until December 31, 1996, 286 patients with non-small cell lung cancer were diagnosed and treated in the Department of Tuberculosis and Respiratory Diseases and Palliative Care Unit in Babice nad Svitavou, Czech Republic. There were 251 (87.8%) males and 35 (12.2%) females. The mean age of the whole group was 63.8 years. According to TNM staging (regarding the locoregional extension of the disease) as the only criterion 112 patients (39.2%) were eligible for surgical resection of primary lung tumour. However lung surgery was performed in 57 patients (19.9%) with successful radical resection in 49 cases (17.1%) out of 286. Surgical procedure was contraindicated in 55 (49.1%) out of 112 potentially resectable patients mostly because of limited cardiopulmonary function. The strategy of neoadjuvant treatment before the resection of lung tumour was used for 12 (4.2%) patients. Chemotherapy, radiotherapy or the combination of both treatment modalities was used in 121 (42.3%) patients. Symptomatic therapy alone was chosen in 104 (36.4%) cases. The performance of radical resection of lung tumours resulted in a significant influence on the overall survival in the subgroup of all patients with potentially locoregionally operable disease according to the TNM staging. The median survival period reached 17 months if radical resection was done compared to 8.5 months if radical resection was not performed. This difference is statistically significant, p = 0.05. PMID- 11382142 TI - [Graves' disease. Clinical features and treatment results]. AB - In the last years the diagnosis of Grave's disease has become easier and accelerated due to new laboratory and instrumental tests. The therapeutic possibilities improved also by using antithyroidal drugs, surgery and radioiodine application subsequently. In a retrospective study 278 outpatients with Graves disease, treated and controlled by our centre for thyroid diseases in 1992-1999, were investigated to evaluate the initial and leading subjective symptoms which led the patients to seek medical help ("guide symptoms"). These obtained guide symptoms were compared with the objective "clinical signs" observed at the beginning of the disease. Weight loss turned out to be the most frequent (26%) guide symptom, whereas tachycardia (> 90 beats/min) and moist skin were the most frequently observed clinical signs (71 and 61% respectively). The beginning of therapy, dosage of drugs, length of application and results of treatment are critically evaluated, with special emphasis on the rate and time of remission, on the frequency of recurrences and on the success of additional treatments such as surgery and/or radioiodine application. Pharmacological treatment alone induced a persistent remission (at least 6 months of euthyroidism) in 156 patients (in 113 after the first and in 43 after the 2nd drug trial). Surgery was successful in 46, radioiodine application in 40 patients. The sequential use of drug therapy, surgery and radioiodine as it is performed in our institution leads to a permanent remission in 242 of 278 (= 87%) patients. PMID- 11382143 TI - The influence of new scientific information on the treatment of elderly patients in general practice. AB - It has been proven that with an increasing number of diseases elderly patients are treated by an increasing number of drugs despite the fact that treatment of elderly persons should be as simple as possible. Simpler dosage of (fewer?) drugs may contribute to a good cooperation on the part of patients, as well as to a lower incidence of undesirable effects and drug interactions. Sticking to established medication schemes is another feature observed in practice, which interferes with the introduction of novelties. The aim of this study was to investigate the actual situation of medication of elderly patients treated by general practitioners in this country. Between 1996 and 1998, a random group of 1481 patients aged older than 75 was studied in cooperation with general practitioners. A detailed history was obtained and physical examination was performed, signs of depression were assessed, Barthel's test of everyday activities, and IADL (activities of daily living) and MMSE (Mini Mental State Examination) tests were applied. A unified "Protocol on Examination" was used in which three questions were concerned with medication--the kinds of drugs taken by the patient, their names and dosage, and whether any hypnotics were taken. The five most frequently prescribed groups include vasodilators (62% patients), cardiotonics (39%), analgetics (41%) and Ca-antagonists (25%). The dynamics of the prescription were remarkable--a significant decrease of the use of analgetics and cardiotonics was observed in comparison with a significant increase in the use of ACE inhibitors. The number of drugs administered is as follows: while only 1.3% patients took no drugs, 1.6% patients took more than 13 drugs. 61% patients rank among the categories taking 4-5 or 6-8 kinds of drugs. On the whole, general practitioners tend to prescribe medicaments in doses one tablet per day. CONCLUSIONS: The results suggest that, even nowadays, elderly patients are treated with a rather high number of medicaments. In contrast, the dosage of one tablet a day decreases the total number of the tablets taken. A significant influence of new scientific information was visible in the prescription habits of general practitioners. Be that as it may, in elderly patients, the simplest possible medication should continue to be aimed at. PMID- 11382144 TI - [Prescription patterns and quality of information provided for consumers of benzodiazepines]. AB - The long-time benzodiazepine use by a considerable part of the population and its adverse consequences such as somatic and cognitive side effects, interactions with other drugs and alcohol and its possible impairment of quality of life has provoked a critical discussion about the practice of prescribing benzodiazepine and the information provided by physicians. We therefore investigated these issues in admitted patients of a general hospital. All patients having been admitted to the department of Internal Medicine and taking benzodiazepine were asked by means of a semi-structured interview, which was repeated after 1 year. In 90% of later benzodiazepine discontinuers and in almost 50% of benzodiazepine continuers, tranquilisers were first prescribed during hospital admission. Only 2% of all patients estimated the information provided by the prescribing physicians as satisfying and about 2/3 reported that they did not get any information at all. 66% of all patients were informed about dependency risks. This information was mainly provided by pharmacies, friends and patient information leaflets; only about 20% of all patients were informed by the prescribing physician. Guidelines for tranquiliser prescription only seem to have a poor impact on the clinical practice. The education of physicians about the risks of low-dose dependencies and especially about complementary therapy methods needs to be improved. PMID- 11382145 TI - Tmolt4 leukemic type II isoform of IMP dehydrogenase as a target for 1,2,4 triazolidine-3,5-diones, 1-(1-(3-methylphenyl)ethylidineamino)-4,4-diethyl-3,5 azetidinediones, 3,5-isoxazolidinediones, and 4,4-disubstituted-3,5 pyrazolidinediones. AB - The 1,2,4-triazolidine-3,5-diones, 1-(1-(3-methylphenyl)ethylidineamino)-4,4 diethyl-3,5-azetidinediones, and 4,4-disubstituted-3,5-pyrazolidinediones proved to be potent competitive inhibitors of human Tmolt4 leukemia Type II IMP dehydrogenase [IMPDH] activity, an enzyme isoform which is induced in highly proliferating cells. On the other hand, the 3,5-isoxazolidinediones were shown to be uncompetitive inhibitors of Type II IMPDH activity. The correlation between inhibition of Type II IMPDH activity with the agents' ability to suppress DNA and purine syntheses in these Tmolt4 leukemia cell was positive. Type I IMPDH (i.e., the isoform that is present in normal cells) was not inhibited by these compounds suggesting that these agents would be less toxic to normal cells and have selective inhibition towards proliferating cells. PMID- 11382146 TI - Synthesis and antimicrobial activity of novel pyrazolo[3,4-b]quinoline derivatives. AB - New pyrazolo[3,4-b]quinoline derivatives have been prepared by cyclization of the intermediate 2-chloroquinoline-3-carbonitrile 7, namely 3-amino-1H-pyrazolo[3,4 b]quinoline 8a, 3-amino-1-phenyl/(p-substituted)phenyl/-1H-pyrazolo[3,4-b] quinoline 8b-f. Furthermore, 3-[(3-aryl-4-oxothiazolidin-2-ylidene)amino]-1H pyrazolo[3,4-b]quinolines 11a,b; 3-[(3-aryl-4-oxothiazin-2-ylidene)amino]-1H pyrazolo[3,4-b]quinolines 12a,b and 3-(2-aryl-4-oxothiazolidin-3-yl)-1H pyrazolo[3,4-b]quinolines 13a,b were synthesized. The antimicrobial activity was evaluated for most of the prepared compounds. PMID- 11382147 TI - Hexamethonium-type allosteric modulators of the muscarinic receptors bearing lateral dibenzazepine moieties. AB - Alkane-bisammonium compounds carrying lateral phthalimido substituents are known to have a high affinity for the allosteric binding site of the acetylcholine M2 receptor. The purpose of this study was to replace the lateral phthalimido moieties with rigid tricyclic skeletons of a large volume in order to learn more about the function of the lateral heterocycles. In addition, methyl groups were introduced into the lateral connecting chains. Allosteric inhibition of the dissociation of [3H]N-methylscopolamine from the M2 receptors in porcine cardiac homogenates served to indicate binding of the test compounds to the allosteric site. The phthalimido groups could be replaced with dibenzazepine moieties without any loss in potency. Interestingly, the additional methyl group in the lateral spacer seems to have a significant influence on the allosteric behaviour. PMID- 11382148 TI - 1-(2,6-dichloro-4-hydroxyphenyl)-2-phenylethanes--new biological response modifiers for the therapy of breast cancer. Synthesis and evaluation of estrogenic/antiestrogenic properties. AB - [meso-1,2-Bis(2,6-dichloro-4-hydroxyphenyl)ethylenediamine]platinum(II) complexes (meso-1-PtLL'; L,L' = Cl or L = H2O and L' = OSO3) are highly effective towards hormone-sensitive, rodent breast cancers due to their significant estrogenic potencies. Their antitumor activities are caused by modification of the immune response. The pharmacophor of these compounds, the 1,2-bis(2,6-dichloro-4 hydroxyphenyl)ethane (23H), was used as the lead structure in a structure activity study with the goal of finding new biological response modifiers for the therapy of breast cancer. As intermediates for the synthesis of the 23H derivatives, the CH3O-substituted stilbenes 12E/12Z-16E/16Z were prepared by reaction of the related benzyltriphenylphosphonium halides with 2,6-dichloro-4 methoxybenzaldehyde by the method of Wittig/Campbell and Donald, respectively. Separation of the E/Z-mixtures was performed by fractional crystallization and/or column chromatography. The E-1,2-bis(2,6-dichloro-4-methoxyphenyl)ethene (17E) was obtained by reductive coupling of 2,6-dichloro-4-methoxybenzaldehyde with TiCl4/Zn according to the method of Mukaiyama. Illumination of the solution of 17E in benzene with UV light resulted in an E/Z-isomerization. Compound 17Z could be isolated from this mixture. The CH3O-substituted stilbenes were transformed into their 1,2-diphenylethanes (12H-17H) by catalytic hydrogenation of the C1=C2 double bond. Ether cleavage of the compounds was performed with BBr3. In the estrogen receptor binding assay all OH-substituted 1,2-diphenylethanes showed affinity to the estrogen receptor, which was about two orders of magnitude lower than that of 17 beta-estradiol. In the uterus weight test on the immature mouse 1,2-diphenylethanes with 4-substituted OH groups proved to be "true" estrogens (19H: 2-F/4-OH; 20H: 2-Cl/4-OH; 23H: 2,6-Cl2/4-OH), while those with a 3 substituted OH group in the 2-phenyl ring showed the properties of a "partial" estrogen (18H: 3-OH) or of an "impeded" estrogen (21H: 2-Cl/3-OH; 22H: 2-Cl/5 OH). The latter also showed significant additional antiestrogenic activity. The related E-stilbenes mostly exhibit similar hormonal activities. As a rule, the replacement of the OH groups by the CH3O groups and the change from the E- to the Z-configuration led to a reduction of the estrogenic potencies. Several of the 1 (2,6-dichloro-4-methoxyphenyl)-2-phenylethenes (12E: 3-OCH3; 12Z: 3-OCH3; 15E: 2 Cl/3-OCH3; 15Z: 2-Cl/3-OCH3; 16E: 2-Cl/5-OCH3) produced antiestrogenic effects in the uterus weight test. It is supposed that those new 1-(2,6-dichloro-4 hydroxyphenyl)-2-phenylethanes endowed with marked estrogenic properties are also active as biological response modifiers in animals bearing hormone-sensitive breast cancer. The antiestrogenic derivatives presumably inhibit the breast cancer development by competing with tumor growth stimulating endogenous estrogens for the binding to the receptor. This is to be confirmed in a further study. PMID- 11382149 TI - Synthesis and evaluation of steroidal hydroxamic acids as inhibitors of P450 17 (17 alpha-hydroxylase/C17-20-lyase). AB - With the aim of developing new inhibitors of 17 alpha-hydroxylase/C17,20-lyase (P450 17, CYP 17), two steroidal hydroxamic acids (compounds 2 and 3) were synthesized and evaluated as inhibitors of CYP 17. The synthesis was performed using carboxylic acids as starting material to give acid chlorides which were reacted with N,N,O-tris(trimethylsilyl) hydroxylamine. Using microsomal fractions of human and rat testes and progesterone as a substrate, both compounds moderately inhibited the human and rat enzyme. PMID- 11382150 TI - [Disease in Velazquez' painting]. AB - A short biographical abstract of Painter Velazquez and his artistical and cultural background with relation to this own time is given. His whole work and of the court and official jobs he had has, been analysed. Together with his artistical vocation and great capacity and aptitude for painting, he felt the ambition to seek a position of nobility. A peculiar research of the pathological aspects appearing in his pictures has been made, regardless of the technical characteristics of his painting and style. For this purpose, his paintings have been classified in still lives, religious pictures, mythological pictures, portraits of royal male and female family members, portraits of ordinary people, big peoples such as Las Lanzas, Las Hilanderas and Las Meninas, etc., to conclude with the really singular group of buffoons, dwarfs, jesters and mad people. The pathological issues have been commented. A list of bibliographical references is also added. PMID- 11382151 TI - [Menisci transplantation and autologous chondrocytes]. AB - We report 66 patients with chondral and osteochondral knee lesions and 3 with ankle lesions in whom, after clinical and radiological examination, arthroscopy was performed to harvest 0.3 or 0.4 g of hyaline cartilage from the knee or ankle to be sent for culture. After culture, autologous chondrocytes were implanted using open surgery. Our statistics show 85-90% of excellent and good results in patients under 50 years of age, including 30 elite athletes who were able to return to sport. The best results in the knee were obtained in the femoral condyle, and in the talus for ankle lesions. In one patient, an ACI in the lateral condyle and a meniscal graft were performed in the same surgery, thus restoring the normal physiology and biomechanics of the joint. PMID- 11382152 TI - [Necrologic session. In memory of the Honorable Mr. Enrique Costa Novella. October 24, 2000]. PMID- 11382153 TI - [The moral conscience in the depressive patient]. AB - The two fundamental forms of the depressive guilt are the feeling of guilt and the delusion of guilt. Both phenomena emanate from a conscience distorted because of the depressive mood by means of an holothymic and catathymic mechanism. Also the other three dimensions of depression (anergy, discommunication and rhythmopathy) take part in producing this moral distortion. The appearance of the depressive guilt happens only when these two factors are present: a previous personality with a great spiritual development and a complete or tetradimensional depression. Shame and distrust replace the guilt in the asiatic and african countries. PMID- 11382154 TI - [Spanish disinfectants for the 21st century]. AB - There are two chemical disinfectants patents from Spain that permit to obtain advantageous products on other disinfectants: Nduopropenide (two iodures of quaternary ammonium) and "Peroxidine" (hydrogen peroxide that active to lactic acid and a surfactant mixture). The first product is used as an antiseptic or disinfectant, but the second, only act as disinfectant. DISINFECTION: It is studied (by germ-carrier methods), the microbicide effect on different microorganisms (Gram positive cocci, Gram negative bacilli, fungus, Mycobacteria and B subtilis spores), comparing these two products with different disinfectants as 2% glutaraldehyde, 1/8 phenate-glutaraldehyde, peracetic acid compounds, 11% oxygen peroxide and 2% sodium hypoclorite. It is obtained that 1/4 Peroxidine in 5 minutes or 1/6 Peroxidine in 10 minutes, are the most effective disinfectant on all microorganisms used (includes the most resistant) since it produces destruction of 4 log-10 of spores and 5 log-10 of Mycobacteria. Moreover, it can destroy, completely, the inoculum of commercial spores, routinely used for sterilization process evaluation, in 20 minutes, when 2% glutaraldehyd needs 3-10 hours. ANTISEPSIE: It is studied the "hygienization" and surgical handwashing with Nduopropenide solution, in comparison with classical washing methods (neutral soap in routinely handwashing and 5% chlorhexidine or 10% iodine povidone in surgical washing): 1) Nduopropenide and alcohol solution is more effective that routinely handwashing. 2) This product is more effective and persistent, after surgical washing that chlorhexidine or iodine-povidone. Moreover, it does not must be applied with brush. 3) The mixture Nduopropenide and chlorhexidine makes a synergy, then it can be used in hand or skin washing, on heath personnel or patient people, being advantageous on the other products. PMID- 11382155 TI - [Genetic alterations in the differential diagnosis of melanocytic diseases]. AB - The biological significance of melanocytic lesions depends on its association with melanomas. Some melanocytic lesions are considered as precursors of melanoma while others may share histological similarities with malignant lesions. Melanomas exhibit clinical, epidemiological and histological heterogeneity. Molecular and cytogenetic studies may supply additional information to supplement the histopathological evaluation. Several genes have been linked to melanomas: CMM1 on chromosome 1p36 and CMM2 on 9p21, and other chromosomal regions where tumor suppressor genes are located as p16 (9p21) and p53 (17p13). We analyzed different melanocytic lesions to determine LOH at 9p21 and 17p13 and to assess clonality by the HUMARA technique. All the malignant melanomas were monoclonal and all the benignant lesions we analyzed were policlonal in our series including deep penetrating nevi and Spitz Nevi. LOH at 9p21 was determined in 60% melanomas, 50% Spitz and 0% intradermal nevi. In conclusion, genotypic changes may supplement the phenotypic or morphological evaluation of melanocytic lesions. PMID- 11382156 TI - [Reversion of cancer and mitochondrial filamentation]. AB - It is concisely described the dedication of the author during the last thirty years to the study of the nature, cause and treatment of cancer through the investigation by his group of the energy metabolism of tumors. It is defined the precise responsibility of the author in the discoveries of the second site of control of glycolysis, rotenone tumors, quelamycin and thioproline, filamentous mitochondria, and the technique of cancer reversal by dual strategy. PMID- 11382157 TI - [Microbiopsy and endometrial dating]. AB - This paper summarizes the experience from 78,000 biopsies of the endometrium taken and studied in collaboration with Dr. Francisco Nogales from 1944 to 1982. This is one of the biggest series existing on this matter and its commentary comprises all the changes in our idea of endometrial biopsy which have taken place in this long period of time. PMID- 11382158 TI - [Thalamo-cortical system and consciousness]. AB - After reviewing the concept of the specific and non specific thalamo-cortical systems, the connectivity of the relay and intralaminar nuclei is analyzed as well as the recent data concerning the chemical identity of thalamic neurones, the concept and distribution of "matrix" and "core" neurones and its functional role. The intrinsic electrical properties of thalamic neurones, its mode of discharge--depending of the membrane potential level--and its functional significance in the context of the brain's global activity are discussed. Of special interest are the studies on the effects of lesion of the relay and intralaminar nuclei as well as its repercussion in the interpretation of the sensory perception. After intralaminar nuclei lesion the individual is not aware of the nformation conveyed through the specific channels. It follows a discussion on the importance of the temporal and spatial mapping in the elaboration of perception and cognition. Due to the intrinsic electrical properties and the connectivity of thalamic neurones two groups of corticothalamic loops are generated, which resonate at a frequency of 40 Hz. The specific thalamo-cortical loops give the content of cognition and the no specific loop, the temporal binding required for the unity of the cognitive experience. Consciousness is then, a product of the resonant thalamo-cortical activity, and the dialogue between the thalamus and cortex, the process that generates subjectivity, the unique experience we all recognized as the existence of the "self". PMID- 11382160 TI - [Sporadic primary hypothyroidism in children yesterday and today]. AB - The evolution of congenital primary hypothyroidism has extremely capitalized on early L-T4 substitutive therapy, as a result of neonatal screening techniques, that allow early diagnosis. However, IQ and neurodevelopment of patients diagnosed and treated in this manner are yet quite different from those normal controls. Latest efforts to achieve precocious neonatal diagnosis have led to earlier initiation of treatments. These circumstances, in addition to the use of higher L-T4 doses (between 10 and 15 micrograms/Kg/day) have permitted to improve clinical responses, in terms of QI and neurodevelopment. Still concern remains about the potential late side effects of high L-T4 doses, that could promote conduct and behavior alterations in these patients in the coming years; in this regard, some suspicions have been shown. PMID- 11382161 TI - [Current status of refractive surgery]. AB - RATIONALE: An attempt is made to review the different surgical procedures for the treatment of the ocular refraction defects and their organs. Special emphasis is placed on the advantages and disadvantages of them, their principal indications and present situation in the surgeon's strategy when solving the problems which, in practice, are caused by these defects, as well as the possibilities that are being tested to solve the problems that presently have not satisfactory response. METHODOLOGY: In addition to analyzing the bibliography, the procedures used by the author and his or her opinion on them, an attempt is made to group them based on the objective on which the intervention is centered to obtain the result desired. COMMENTARY: Both the accuracy of the procedures as well as the complications that are presently derived from this surgery are discussed. Reasonable limits of each procedures, the reasons that make it obligatory to discontinue some of them and the problems that must be solved in the future to up date each one of the techniques presently in use are commented on. PMID- 11382162 TI - [A case of fever of unknown origin]. PMID- 11382163 TI - [Circadian periodicity in human heart rate daily variability disorders: subsequent demonstrations through a chronobiological approach to the values of estimated alpha exponent on sinusal R-R intervals]. AB - OBJECTIVE: The present study is aimed at giving a further demonstration that the disorder in human heart rate variability is characterized by a circadian periodicity. Such an evidence has been provided by estimating the alpha exponent of the Levy's stable distribution approximated to the histogram of the differences between the durations of two adjacent sinusal R-R intervals. MATERIALS AND METHODS: The purpose has been pursued by estimating the alpha exponent on fragments of one hour of the Holter electrocardiogram executed in 10 clinically healthy subjects (5 males and 5 females, ranging in age from 23 to 30 years). RESULTS: The chronobiological analysis of the hourly-qualified values of the alpha exponent has demonstrated a significant circadian rhythm for this coefficient. CONCLUSIONS: The significant circadian periodicity of the alpha exponent is a further evidence that the disorder in human heart rate variability, herein represented by the extreme variability of the differences between the durations of two consecutive sinusal R-R intervals, is characterized by a periodic recursivity. PMID- 11382164 TI - [Partially hydrolyzed guar gum: a fiber as coadjuvant in the irritable colon syndrome]. AB - PURPOSE: Partially hydrolyzed guar gum (PHGG) is a water-soluble dietary fiber, possessing non-gelling properties. The objective of this clinical experience was to evaluate the progress of symptoms and the modifications in the frequency of evacuation in subjects affected by IBS and regularly taking PHGG. PATIENTS AND METHODS: The group was made up of 134 out-patients of both sexes, average age 43.12, suffering from IBS, both obese and of normal weigh, with a mean number of weekly evacuations between 2 and 35. The subjects, divided in 2 groups on the basis of Body Mass Index (BMI), were submitted for 24 weeks to a balanced, low or normal calorie diet supplemented by 5 g a day of PHGG. The following information was gathered: number of weekly evacuation, typical symptoms of IBS, cholesterol, triglycerides and glucose levels. In a few subjects (n. = 34) also the plasmatic electrolyte levels, before and during PHGG intake, were evaluated. RESULTS: Both groups showed positive results in the evacuation frequency (p < 0.01 at 12th week) and a decrease, after 3 weeks of PHGG intake, in frequency of IBS symptoms such as flatulence (-55.6%), abdominal tension (-4.7%) and abdominal spasm ( 35%). On the other hand an increased number of subjects showed normal levels of cholesterol (+12.2%), lipids (+26.9%) and glucose (+16%). Concentrations of plasmatic electrolytes didn't change during PHGG intake, except for a marked increase of selenium levels, compared to pre-intake levels. CONCLUSIONS: The observations obtained from this clinical experience reassert that dietary fiber supplementation is useful in cases of altered intestinal motility. PHGG, due to its water-solubility and non-gelling properties, can be useful also in IBS. PMID- 11382166 TI - [Assessment of teaching quality: from normative aspects to the relapse of the formative project]. AB - INTRODUCTION: To base the medical student's education on scientific evidence, we need to applied to medical education the same evidence-based methods characteristics of scientific research. Our goal is to change curricula, educational methods, teaching of clinical skills, in order to improve professional training of medical and nursing students. Our work highlights the student's point of view relative to changes of educational project, that is a constitutive aspect of best evidence medical education. DESIGN: Every year, an evaluation test is submitted to all medical, nursing and nutritionist students of Universita "Campus Bio-Medico". This test worked out by both teachers and students, is designed to explore student's perception of all aspects, educational and relational, related to the university. Data are been processed using explorative analysis of principal elements, and then using factorial analysis with "Varimax", data orthogonal rotation. A specific database in Microsoft Access, is been used for data entry, while statistical analysis is been performed using didactic software STATA (Stata Corporation). RESULTS: According to data, we can claim that our students evaluate their teachers and tutors depending on two principal factors: on one hand educational skills, that include personal competence on teaching and getting in touch with the students; on the other hand managing and planning skills. These are very important to overcome the dangers related to integrated courses, composed by many different scientific matters and planned by many teachers: without a very good planning, students may not be allowed to achieve clear, synthetic and well-structured knowledge. CONCLUSIONS: Students want to be regarded as adult learners, they wish to achieve a well structured knowledge, both composed by theoretical and practical skills and personal relations, in order to think of every activity according to an organic knowledge. PMID- 11382165 TI - [Epilepsy and pregnancy]. AB - Epilepsy is one of the most common pre-existing neurologic disorders encountered by the obstetrician (1) and pregnant women with epilepsy constitute 0.5% of all pregnancies (2). Pregnant women with epilepsy are at risk for a variety of complications (3). Pregnancy may influence the course of epilepsy and epilepsy and antiepileptic drug therapy may have an impact on the course of pregnancy and pregnancy outcome. This study represents a retrospective analysis of 40 pregnant patients with epilepsy. PMID- 11382167 TI - [Vaccine shortage and re-emergence of influenza. The problem in the USA]. PMID- 11382168 TI - [Evidence-based medicine: accurate review of the international literature]. AB - The Authors emphasize the continual scientific technological advances of medical science, the appearance of new nosological extents and the increased health request by people. Physicians are called to up-to-date all the time to keep a high-level of professional competence also after the end of the university studies. Every medical staff can use EBM, setting aside all personal specialization and also with the limitations estimated in our research. PMID- 11382169 TI - [Second-line chemotherapy and new drugs for ovarian carcinoma]. AB - Epithelial ovarian carcinoma represents a prototypic example of neoplastic disease sharing at the same time good chemosensitivity as well as marked propensity to relapse. If in one hand the definition of almost clear-cut guidelines has been reached in the setting of first-line therapy (i.e., cytoreductive surgery followed by first-line chemotherapy), more difficult to discern for the oncologist remains the choice of treatment in the occasion(s) of relapse(s). This article focuses on this particular setting of disease, analyzing the specific criteria of choice of drug or their combinations; the definite criteria followed for the use of platinum compounds in the second line of treatment, even if already utilized in the front line therapy, are also analyzed. Specific attention has also been paid in the definition of the role of single parameter as a "prognosticator" and/or as "predictor" of response. The Authors emphasize how clinical definitions remain the more reliable, simple, reproducible tools in therapeutical decision making. PMID- 11382170 TI - [A 35-year-old woman with fever, dyspnea, and pain in the left thigh]. AB - A thirty-five years old woman during her twelfth pregnancy presented fever and pain at the left thigh. After cesarean delivery dyspnea added to the first two symptoms and pulmonary embolism was suspected. A clinical history revaluation suggested a diagnosis of infectious endocarditis and femoural osteomielitis due to a septic embolus. PMID- 11382171 TI - [Successful cancer chemotherapy. The cure of Hodgkin's disease. III]. PMID- 11382172 TI - [LDL-apheresis: current status]. AB - PURPOSE: State of the art of LDL-apheresis and treatment of severe familiar hypercholesterolemia. PATIENTS AND METHODS: Clinical experience of treatment with LDL-apheresis of pediatric and adult patients with familial hypercholesterolemia using the following techniques: D.A.L.I., H.E.L.P., D.S.C. RESULTS: The outcome of treatment with LDL-apheresis in young patients, using the most recently introduced techniques, is reported. We have submitted to LDL-apheresis 11 pediatric patients. The youngest is aged 3.5. LDL-apheresis is able at improving the metabolic impairment and halting the natural evolution of atherosclerotic disease and atherosclerotic complications. CONCLUSIONS: At present, LDL-apheresis is the most effective and safe therapeutic approach to the treatment of homozygous, heterozygous and double heterozygous familial hypercholesterolemia. PMID- 11382173 TI - [Counseling of immigrant families]. AB - The quality and efficiency in the process of psychological counseling with migrant families depends on the counselor's insight in the constellations of migration and culture. Both have a special influence on the way of coping with problems and the relation between client and counselor during the therapy. It is therefore necessary to adapt the own therapeutical approach to the migrant's experiences and abilities of comprehension. The author presents one example of intercultural family counseling by three short case explorations. PMID- 11382174 TI - [Counseling of socially disadvantaged families]. AB - Poverty includes more and more families with children. In Germany nearly every fifth child is affected by poverty. Poverty is a very hard risk for the development of children. The biographies of family living in poverty are reconstructed by representative materials of an educational counseling office in Munich which is engaged with social discrimination for 23 years. Studies of resilience are especially suited to explain the psychic impacts of poverty for children. In conjunction with the community psychology they offer impulses for the arrangement of efficient help. Finally the author presents different consulting strategies of conduct and practical examples of individual help. PMID- 11382175 TI - [Cooperation--a prerequisite strategy in the psychosocial practice]. AB - With the help of empirical data requirements of successful cooperation will be described. At first concepts are clarified, then theoretical assumptions and empirical results are reported and at least on the basis of own empirical data conditions for cooperation are developed. PMID- 11382176 TI - [Delivery of counselling to divorcing families--new ways of interdisciplinary cooperation for the benefit of the child]. AB - The essence of the reformed German law of parent and child (Kindschaftsrecht) for families who have filed for divorce is that parents need counselling and assistance rather than patronizing from the state when it comes to decisions regarding the custody of children. Prior to the passing of the new law, critics doubted whether parents would of their own accord avail of the services of youth counselling institutions. A survey of all youth welfare offices in Bavaria documents practices since the law was revised. The results indicate that closer cooperation between the parties involved is of vital importance and that the youth welfare offices must invest a great deal of energy in public relations in order to encourage families to avail of counselling. A pilot project, in which family courts and youth welfare offices work in close cooperation, illustrates how this might function. PMID- 11382177 TI - Dioxin in Vietnam--the ongoing saga of exposure. PMID- 11382178 TI - Recent dioxin contamination from Agent Orange in residents of a southern Vietnam city. AB - Marked elevation of dioxin associated with the herbicide Agent Orange was recently found in 19 of 20 blood samples from persons living in Bien Hoa, a large city in southern Vietnam. This city is located near an air base that was used for Agent Orange spray missions between 1962 and 1970. A spill of Agent Orange occurred at this air base more than 30 years before blood samples were collected in 1999. Samples were collected, frozen, and sent to a World Health Organization- certified dioxin laboratory for congener-specific analysis as part of a Vietnam Red Cross project. Previous analyses of more than 2200 pooled blood samples collected in the 1990s identified Bien Hoa as one of several southern Vietnam areas with persons having elevated blood dioxin levels from exposure to Agent Orange. In sharp contrast to this study, our previous research showed decreasing tissue dioxin levels over time since 1970. Only the dioxin that contaminated Agent Orange, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), was elevated in the blood of 19 of 20 persons sampled from Bien Hoa. A comparison, pooled sample from 100 residents of Hanoi, where Agent Orange was not used, measured blood TCDD levels of 2 parts per trillion (ppt). TCDD levels of up to 271 ppt, a 135-fold increase, were found in Bien Hoa residents. TCDD contamination was also found in some nearby soil and sediment samples. Persons new to this region and children born after Agent Orange spraying ended also had elevated TCDD levels. This TCDD uptake was recent and occurred decades after spraying ended. We hypothesize that a major route of current and past exposures is from the movement of dioxin from soil into river sediment, then into fish, and from fish consumption into people. PMID- 11382179 TI - Health surveillance for occupational chronic obstructive pulmonary disease. AB - Screening for lung disease in workers with a low prevalence of chronic obstructive pulmonary disease (COPD) is subject to debate. Examining all workers would lead to unacceptable costs. Surveillance of high-risk groups may be a useful alternative. Data from 314 workers exposed to dust were used to develop a prediction model. Data from a study comprising 96 rubber workers were used to validate the model's performance. COPD was defined as a ratio of forced expiratory volume in 1 second to forced vital capacity outside the 5th percentile. The accuracy of the model was evaluated by receiver operating characteristic curve analysis and by comparison of the observed versus predicted probabilities. Shortness of breath, wheeze, heavy smoking, and work-related lower respiratory symptoms were identified as independent determinants of having COPD. Workers with COPD can be distinguished from those without COPD on the basis of a medical/work-related questionnaire. Occupational physicians can develop a low cost strategy for detecting workers at risk for COPD. PMID- 11382180 TI - Predictive validity of the Strain Index in turkey processing. AB - The Strain Index is a job analysis method for determining if workers are exposed to increased risk of developing distal upper extremity disorders. Its predictive and external validity was initially demonstrated in a pork processing plant. The purpose of this study was to evaluate the predictive validity of the Strain Index in one turkey processing plant. While blinded to health outcomes, investigators analyzed the right and left sides of workers in 28 jobs using the Strain Index and classified them as "hazardous" or "safe" based on the Strain Index score. Subsequently, OSHA 200 logs were used to ascertain the occurrence of distal upper extremity disorders retrospectively. If at least one such disorder had occurred on the right or left side during the previous 3 years, that side was classified as "positive." If no such disorder was reported during the previous 3 years, that side was classified as "negative." When comparing sides, symmetry between morbidity and hazard classification was required. When comparing jobs, such symmetry was not required. Evidence of association between the hazard classifications and the morbidity classifications for the 56 sides and the 28 jobs was evaluated using 2 x 2 contingency tables. For the sides, the association between hazard classification and morbidity classification was statistically significant, with an odds ratio of 22.0. The sensitivity, specificity, positive predictive value, and negative predictive value were 0.86, 0.79, 0.92, and 0.65, respectively. Similar results were noted for the jobs--the odds ratio was 50.0, and the sensitivity, specificity, positive predictive value, and negative predictive value were 0.91, 0.83, 0.95, and 0.71. These results provide additional evidence of the external validity and predictive validity of the Strain Index. PMID- 11382181 TI - The utility of Poison Control Center data for assessing toxic occupational exposures among young workers. AB - This study assessed the utility of Poison Control Center data for identifying and describing occupational toxic exposures to youth and the associated health outcomes of such exposures. The authors reviewed 1997 data from the American Association of Poison Control Centers' Toxic Exposure Surveillance System for occupational exposures to persons 6 to 19 years old. In 1997, there were 3442 reported occupational exposure cases among 6- to 19-year-olds. Exposure cases included underage workers, exposure to multiple substances, continuous or repeated exposure to the offending substance, and exposures occurring in the home. Poison Control Centers provide unique and valuable information on toxic workplace exposures to youth. Child labor laws may be violated in many cases, and risks exist with respect to industrial work done in the home. Prospective studies with immediate follow-back are needed. PMID- 11382182 TI - Adverse respiratory effects following overhaul in firefighters. AB - Overhaul is the stage in which firefighters search for and extinguish possible sources of reignition. It is common practice not to wear respiratory protection during overhaul. Fifty-one firefighters in two groups, 25 without respiratory protection and 26 wearing cartridge respirators, were monitored for exposure to products of combustion and changes in spirometric measurements and lung permeability following overhaul of a structural fire. Testing at baseline and 1 hour after overhaul included forced vital capacity (FVC), forced expiratory volume in one second (FEV1), serum Clara cell protein (CC16), and serum surfactant-associated protein A (SP-A). Overhaul increased CC16 in both groups, indicating increased alveolarcapillary membrane permeability. Contrary to expectations, SP-A increased and FVC and FEV1 decreased in the firefighters wearing cartridge respirators. Changes in FEV1, CC16, and SP-A were associated with concentrations of specific products of combustion or carboxyhemoglobin levels. Firefighter exposures during overhaul have the potential to cause changes in spirometric measurements and lung permeability, and self-contained breathing apparatus should be worn during overhaul to prevent lung injury. PMID- 11382183 TI - Abnormal liver function associated with occupational exposure to dimethylformamide and hepatitis B virus. AB - N,N-Dimethylformamide (DMF) has excellent solvent properties and is used intensively in the production of synthetic leather and resins. It has caused hepatoxicity in human and animal studies. Hepatitis B virus (HBV) and hepatitis C virus infections are reported to be the major causes of chronic liver diseases (including liver cirrhosis and liver cancer) in Taiwan. This study examined the dose-response relationship of the observed abnormal liver function among the DMF exposed workers and the interactions among DMF, other chemical exposures, HBV infection, and potential confounders on liver abnormalities. The average DMF exposure concentration was 11.6 ppm (median, 5.9 ppm; range, 0.1 to 86.6 ppm); 65 of 176 workers (36.9%) had high (> 10 ppm) DMF exposure, 37 (21%) had middle (> 5 ppm, < or = 10 ppm) exposure, and 74 (42%) had low (< or = 5 ppm) exposure. There were 24 of 65 abnormal liver function test results (LFTs) (36.9%) (elevations of either glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, or gamma-glutamyl transpeptidase) among the workers with high DMF exposure, 10 of 37 abnormal LFTs (27%) among workers with middle DMF exposure, and 11 of 74 abnormal LFTs (22%) among workers with low DMF exposure. Compared with the workers having low DMF exposure, the HBV, drinking, body mass index (BMI), sex, duration of employment, epichlorohydrin, and toluene exposure adjusted odds ratios (ORs) (and 95% confidence intervals [CIs]) for abnormal LFTs were 1.62 (0.61, 4.28) for workers with middle DMF exposure and 2.93 (1.27, 6.8) for those with high DMF exposure, and there was a significant dose response between DMF exposure and the prevalence of abnormal LFTs (P = 0.006). There were significant associations between abnormal LFTs and HBV carriers (adjusted OR: 3.11; 95% CI: 1.29, 7.5; P = 0.01) and between abnormal LFTs and increased BMI (adjusted OR: 2.2; 95% CI: 1.02, 4.72; P = 0.041). Ultrasonography showed significant associations between chronic liver diseases and HBV carrier status, increased BMI, and high cumulative (> 100 ppm-years) DMF exposure (respectively, adjusted OR: 9.58, 95% CI: 1.79, 51.4, P = 0.007; adjusted OR: 13.2, 95% CI: 1.32, 132, P = 0.025; and adjusted OR: 6.2, 95% CI: 1.14, 34.1, P = 0.032). Drinking and BMI were significantly associated with fatty liver (respectively, adjusted OR: 4.9, 95% CI: 1.39, 17.3, P = 0.012; and adjusted OR: 7.93, 95% CI: 1.6, 39.3, P = 0.01). In conclusion, this study demonstrated that (1) a significant dose-response relationship existed between liver function abnormalities and DMF exposure among workers in Taiwan, (2) HBV carrier status or increased BMI had synergistic effects with DMF in causing liver abnormalities (abnormal LFTs and clinical chronic liver diseases). PMID- 11382184 TI - Mortality at an automotive engine foundry and machining complex. AB - Mortality was analyzed for an automotive engine foundry and machining complex, with process exposures derived from department assignments. Logistic regression models of mortality odds ratios (ORs) were calculated for 2546 deaths, and numbers of work-related deaths were estimated. Lung cancer mortality in the foundry was increased where cleaning and finishing of castings was performed (OR, 1.7; 95% CI, 1.15 to 2.4 [at mean exposure duration of exposed cases]) and in care-making after 1967 (OR, 1.5; 95% CI, 1.11 to 2.0). Black workers had excess lung cancer mortality in machining heat-treat operations (OR, 2.5, 95% CI, 1.4 to 4.3) and excess nonmalignant respiratory disease mortality in molding (OR, 2.5; 95% CI, 1.16 to 5.5) and core-making (OR, 2.7; 95% CI, 1.25 to 5.8). Stomach cancer mortality was elevated among workers with metalworking fluid exposures in precision grinding (OR, 2.4; 95% CI, 1.14 to 5.1). Heart disease mortality was increased among all workers in molding (OR, 1.6; 95% CI, 1.09 to 2.3), as was stroke mortality among workers exposed to metalworking fluids (OR, 1.8; 95% CI, 1.22 to 2.7). Malignant and nonmalignant liver disease mortality was elevated in assembly/testing and precision grinding. In this modern foundry, 11% of deaths were estimated to be work-related despite it's being largely in regulatory compliance over its 40-year existence. Machining plant exposures accounted for 3% or more of deaths there. PMID- 11382185 TI - Intervention study on acquired color vision deficiencies in styrene-exposed workers. AB - The main aim of the study was to examine the possible effects of occupational exposure to styrene on color vision function and the course after reduction of exposure. Color vision function was examined in 22 styrene-exposed laminators and 11 control subjects at a boat manufacturing plant. The Lanthony D-15 desaturated panel was used to test acquired dyschromatopsia. In all, six examinations were performed: Monday morning and Thursday afternoon of the same week, before and immediately after a vacation of 4 weeks (altogether, phase 1), and approximately 10 months later (phase 2), after the exposure level of styrene had been reduced. Styrene uptake was objectified by biological monitoring measuring the metabolites mandelic acid and phenylglyoxcylic acid in urine samples taken on Thursday afternoon. In both Thursday examinations, styrene-exposed workers had higher color confusion index (CCI) values compared with controls, which indicated quantitative color vision loss. After an exposure-free period of 4 weeks, a significant decrease of CCI values to normal range was found in laminators. Reexamination 10 months later showed also lower CCI values in exposed workers, indicating a dose-effect relationship. Abnormal CCI values occurred primarily in subjects with an excretion of approximately 500 to 600 mg mandelic acid plus phenylglyoxcylic acid per gram creatinine or more. We concluded that styrene induced color vision dysfunction is reversible after an exposure-free interval of 4 weeks. The current Biological Tolerance Value of 600 mg mandelic acid plus phenylglyoxcylic acid per gram creatinine, as used in Germany, protects styrene exposed workers from this subclinical effect. PMID- 11382186 TI - External Practicum-Year Residency Training in Occupational and Environmental Medicine: the University of Pennsylvania Medical Center Program. AB - We describe a competency-based training program that allows physicians employed full-time in occupational and environmental medicine to satisfy the supervised practicum year of training required by the American Board of Preventive Medicine (ABPM). The program is designed for trainees with greater clinical experience than the 1 clinical year required by the ABPM. To date, 25 physicians from clinic based, academic, corporate, and government employment across most geographic regions of the United States have been admitted into the program. Most completed a master's in public health (MPH) in a distance-learning, on-job, on-campus, or executive program. The practicum-year training has been highly successful, as evidenced by improvements in resident self-assessment of competency, resident satisfaction with the training, faculty evaluation of resident performance, and success rate in the ABPM examination. The program has opened a new pathway for physicians making a mid-career shift to occupational and environmental medicine to obtain high-quality, in-depth education and board certification. PMID- 11382187 TI - Mitochondrial encephalomyopathies: therapeutic approaches. AB - Therapy of mitochondrial encephalomyopathies (defined restrictively as defects of the mitochondrial respiratory chain) is woefully inadequate, despite great progress in our understanding of the molecular bases of these disorders. We review available and experimental therapeutic approaches, which fall into seven categories: (1) palliative therapy; (2) removal of noxious metabolites; (3) administration of artificial electron acceptors; (4) administration of metabolites and cofactors; (5) administration of oxygen radical scavengers; (6) gene therapy; and (7) genetic counseling. Progress in each of these approaches provides some glimmer of hope for the future, although much work remains to be done. PMID- 11382188 TI - Molecular basis for treatment of mitochondrial myopathies. AB - Mitochondrial DNA (mtDNA) is the only autonomously replicating source of DNA outside the nucleus. The mitochondrial genome encodes thirteen essential polypeptides of the mitochondrial respiratory chain. Defects of the mitochondrial genome can cause severe neurological and multi-systemic disorders. In many patients there is a mixture of mutated and wild-type mtDNA in the same cell (a situation termed heteroplasmy). In these patients the ratio of mutated to wild type mtDNA is crucial and a biochemical defect only occurs with relatively high levels of mutated mtDNA within an individual cell. This threshold also seems to be critical in the development of mtDNA disease. Since the genetic defect causes a dysfunction in the terminal stage of oxidative metabolism, there is little potential for pharmacological intervention. Molecular techniques must be developed to reverse the ratio of mutated and wild-type mtDNA. In this paper we summarise our approach using both antigenomic peptide nucleic acids and cell necrosis. PMID- 11382189 TI - Molecular basis of treatment in motor neurone disease. AB - The pathways leading to motorneuron degeneration in amyotrophic lateral sclerosis (ALS) are complex. Excitotoxicity, oxidative damage and, maybe, abnormal aggregation of neurofilaments are key events on which therapeutical strategies can be designed. This paper reviews current knowledge on these strategies. Even though we should be aware that appropriate management of disease symptoms remains the most effective therapeutical intervention, understanding the pathophysiology of ALS is essential for developing new therapies. PMID- 11382190 TI - New therapies in muscular dystrophies. AB - Muscular dystrophies are a group of hereditary muscle disorders that often result in severe disability. Curative therapy is not yet available for muscular dystrophies (MD). In the near future, it is not expected that gene-replacement therapy will be available. Other strategies to decrease the rate of muscle necrosis and to increase strength in patients are necessary. Therefore the interest in symptomatic drug treatment has recently increased. A few trials have been performed on different types of muscular dystrophies, and some have generated positive results on muscle strength or muscle mass. We review the state of the art in therapy of MD and summarize the drugs that have been used and the evidence and results of such clinical trials. PMID- 11382191 TI - Retroviral vectors for gene therapy of Duchenne muscular dystrophy. AB - The aim of this review is to summarize the state of art of retroviral vectors for gene therapy of Duchenne muscular dystrophy (DMD). Actual knowledge on this matter indicates that retroviral vectors are able to transduce muscle satellite cells in vivo and that these cells can participate in muscle repair processes, even if the efficiency of transduction of satellite cells remains the limiting factor. Such a process is reminiscent of the normalization process of DMD carriers and offers hope for the development of long-term gene therapy of this disease. PMID- 11382192 TI - Structural comparison of actin binding in utrophin and dystrophin. AB - Dystrophin links the actin cytoskeleton to the dystroglycan complex in the plasma membrane as part of the linkage between the cytoskeleton and the extracellular matrix. Damage to or absence of dystrophin causes Duchenne or Becker muscular dystrophy. It has been proposed that elevating the levels of utrophin, a close homologue of dystrophin, may act as a therapy for these forms of muscular dystrophy. This requires that there is a close functional equivalence of these two proteins. In both utrophin and dystrophin, the main actin-binding region is at the N terminus. It is related to sequences found in a number of other proteins including alpha-actinin, spectrin and fimbrin. Recent structural and biochemical studies of these proteins have shown that although the method of binding to actin is broadly similar, there are significant differences. There are even differences between utrophin and dystrophin. These studies imply that some caution should be applied to claims that utrophin and dystrophin are completely functionally interchangeable. In this paper, I review studies that elucidate and compare the actin-binding function of utrophin and dystrophin, particularly those initiated in the laboratory of Dr. John Kendrick-Jones at the MRC in Cambridge. PMID- 11382193 TI - The current status of myoblast transfer. AB - As a means of correcting the defect in the muscles of Duchenne muscular dystrophy (DMD) patients, myoblast transplantation has the advantage of utilizing normal biological mechanisms of repair of this tissue. Thus, it has the potential not only for correcting or complementing the genetic defect, but also for restoring the structure and function of pathologically damaged muscle tissue. However, it is also subject to many of the problems of other genetic therapies for muscle tissue, especially the problem of dispersing the therapeutic agent throughout the muscle mass and that of immune rejection of the genetically corrected tissue. In this article, these problems are discussed. In addition the history of cell transplantation therapy for DMD is presented as an example of the practical difficulties that arise from over eager application to human trials of preliminary data from animal experiments. Recent demonstrations that stem or early precursor cells from a muscle and from the bone marrow are able to disperse to sites of muscle damage via the blood vascular system and to participate in muscle repair raise the hope that this mode of therapy may be applicable to muscular dystrophies. PMID- 11382194 TI - Regenerative capacity of human satellite cells: the mitotic clock in cell transplantation. AB - In this communication, we will review the problems caused by cell-mediated gene therapy, taking skeletal muscle as a physiological model. In particular we have utilised vectors transferring telomerase under the control of retroviral promoters into human satellite cells. The set of results presented here has several implications regarding gene therapy trials. Nevertheless, more experiments will be required to fully validate this cellular model and to use telomerase to safely extend the lifespan of putative gene therapy vectors. PMID- 11382195 TI - Therapy in myotonic disorders and in muscle channelopathies. AB - Myotonia and muscle weakness are cardinal features of myotonic disorders including the myotonic dystrophies and the non-dystrophic myotonias. Despite the recent progress in molecular genetics of these myotonic disorders, the precise mechanisms responsible for myotonia and for permanent or episodic muscle weakness are still unclear. Treatment has been mostly symptomatic, independent of the disease process involved. Moreover, there have been few randomized controlled trials of treatment for myotonic disorders and consequently no standardized treatment regimens are available. We present a review of selected treatment trials in the myotonic disorders and in muscle channelopathies, and discuss, on the basis of our experience in the myotonic disorders, the limits and advantages of treatment trials in this field. Future genotype-phenotype correlations using the patch-clamp technique are also illustrated. PMID- 11382196 TI - Cytogenetic damage in peripheral lymphocytes of mitochondrial disease patients. AB - Recent studies indicate an important role of endogenous oxidative stress in the onset and/or in the progression of mitochondrial encephalomyopathies. In particular, the increased production of radical species caused by altered mitochondrial functionality could affect both mitochondrial and nuclear DNA integrity. We performed the micronucleus assay coupled with fluorescence in situ hybridization (FISH) to detect chromosome damage in peripheral blood lymphocytes in a group of patients affected by different forms of mitochondrial encephalomyopathies. Moreover the comet assay has been carried out to detect primary and oxidative damage in the nuclear DNA. Our results show a significant presence of both DNA and chromosome damage in patients compared to a matched group of controls. A reduction in DNA alterations is also observed in patients after treatment with coenzyme-Q10. PMID- 11382197 TI - Gene transfer in skeletal muscle by systemic injection of DODAC lipopolyplexes. AB - Lipid-based vectors are a promising tool for gene therapy applications. Several studies have reported their use in vivo to transfect different organs. Few data, however, are available about lipid-mediated gene transfer in skeletal muscle. Here we report the initial results obtained after systemic administration of lipopolyplexes based on the DODAC cationic lipid in an animal model of muscle regeneration. In particular, we compared three routes of administration: intravenous (i.v.), intracardiac (IC) and intra-arterial (IA). Analysis of reporter gene expression (luciferase) showed that regenerating muscle is more efficiently transfected in all cases and that IA injection is by far the best approach. PMID- 11382198 TI - Quantitative myotonia assessment: an experimental protocol. AB - Severe clinical myotonia can be physically disabling and socially impairing but as yet there is no standardized treatment regimen. The aim of our study is to present a protocol to measure myotonia using quantitative muscle assessment measures. The proposed protocol addresses two main issues. Muscle strength is assessed in 8 muscles on the right and on the left using a myometer (QMA, quantitative muscle assessment) and by testing strength manually using the 5 point MRC scale (5 = normal) in 15 muscles on the right and on the left. Grip myotonia is assessed by: (a) measuring 1/2 and 3/4 relaxation times (RT) after maximum voluntary contraction (MVC) using QMA apparatus; (b) functional tests (time to open a fist 10 times, time to open and squeeze the eyes 10 times, time to climb 10 steps starting from a seated position, time to protrude the tongue 10 times, time to step onto a chair 10 times; (c) subjective measures of the severity of myotonia using an arbitrary 4-point scale (0 = absent, 4 = severe); and (d) electromyography (EMG) relaxation times after MVC. Although QMA seems to be a reliable tool to measure myotonia, there are still a number of unsolved issues. Further studies are needed to ensure the ability of QMA to quantify myotonia and to guarantee the reliability of the results for clinical research purposes. PMID- 11382199 TI - Correlation of clinical function and muscle CT scan images in limb-girdle muscular dystrophy. AB - Knowledge of selective muscle-involvement is useful to the clinician. Therefore we investigated in sarcoglycanopathy and dysferlinopathy: (i) the correlation between clinical scale, MRC scale and CT findings and (ii) the muscles involved. Patients with a definite diagnosis of dysferlinopathy and sarcoglycanopathy were tested for their clinical functions and muscle strength and assigned a functional grade. Nineteen muscles were evaluated by CT. In dysferlinopathy, distal lower limb muscles are involved, while in sarcoglycanopathy proximal muscles are more affected. In both groups, muscles with MRC < 3 often had severe abnormalities on CT imaging, while muscles with MRC > 4 could be either normal or abnormal. Dysferlinopathy and sarcoglycanopathy have different and selective muscle involvement. This may affect certain types of functional activity. There is a possible relationship between CT findings and MRC grade in muscles with a low (< 3) MRC grade. Abnormality on CT scan may precede the clinical symptoms. PMID- 11382200 TI - Potential involvement of ubiquinone in myotonic dystrophy pathophysiology: new diagnostic approaches for new rationale therapeutics. AB - An impairment of mitochondrial function may contribute to the pathophysiology of myotonic dystrophy (MyD). Coenzyme Q10 (CoQ10) deficiency has been previously observed, even if in a restricted sample of patients. The aim of this investigation was to obtain more information about coenzyme Q10 and its relationships to the aerobic metabolism in a group of MyD patients. Serum CoQ10 appeared significantly reduced with respect to normal controls: 0.93 +/- 0.22 vs. 1.58 +/- 0.28 micrograms/ml (p < 0.05). Moreover, the results demonstrated an inverse tendency between CoQ10 levels and the CTG expansion degree. Basal blood lactate levels were significantly higher than controls (p < 0.05). A borderline inverse correlation between CoQ10 and lactate, corresponding to lactate threshold, was found. These data suggest a possible role of CoQ10 in the pathogenesis of MyD, which may be mediated by mechanisms of cellular damage common to the oxidative pathway. Therapeutic strategies may be devised by virtue of this rationale. PMID- 11382201 TI - Long-term treatment with idebenone and riboflavin in a patient with MELAS. AB - We report a patient with MELAS treated for 24 months with idebenone and riboflavin, during which no stroke-like episodes occurred. Moreover neurological symptoms clearly improved, and a recovery of brain MRI and EEG abnormalities was observed. We conclude that the combined treatment with idebenone and riboflavin may restore the metabolic impairment in MELAS, possibly improving the long-term prognosis in these patients. PMID- 11382202 TI - A novel mtDNA mutation in the ATPase6 gene studied by E. coli modeling. AB - This study aimed to understand the pathogenesis of a new mtDNA-related etiology of Leigh syndrome. We identified the T9176G mutation as the molecular basis of Leigh syndrome in a child and looked for alterations in cellular ATP production. We then modeled the new mtDNA mutation in E. coli and analyzed ATP synthesis, hydrolysis, and the ability of the mutated enzyme to pump protons. Our results suggest that the T9176G change results in a novel, fully assembled enzyme which inhibits the holoenzyme probably by blocking the proton pathway. PMID- 11382204 TI - Classroom strategies used by teachers of students with attention deficit hyperactivity disorder. AB - Pediatric therapists working in schools commonly function as consultants to assist teachers to develop strategies to help promote the classroom performance of students with attention deficit hyperactivity disorders (ADHD). This study surveyed general education teachers in northern New England to determine the classroom strategies teachers commonly use and perceive as being effective for improving the performance of children with ADHD. The strategies receiving the highest frequency and effectiveness ratings were enforcing routine and structure, frequent contact, preferential seating, use of motor breaks, and teaching self monitoring of behaviors. The strategies receiving the lowest effectiveness ratings were peer tutoring, timeout, and giving assistance during transitions. Cross tabulations with chi-square analysis of the effectiveness ratings by grade level indicated the ratings for 3 of the 15 strategies, use of a designated quiet space, use of motor breaks, and sensory modulation techniques, were dependent on grade level. Qualitative analysis of teacher responses to open-ended questions provided recommendations for improving the education of children with ADHD, including: increasing special education support in the classroom, smaller class sizes, frequent parent-teacher collaboration, and more hands-on learning experiences. Implications of the results of the study for therapists working with students with ADHD and their teachers are discussed. PMID- 11382203 TI - Abnormal levels of human mitochondrial transcription factor A in skeletal muscle in mitochondrial encephalomyopathies. AB - Human mitochondrial transcription factor A (hmtTFA or Tfam), a 25-kDa protein encoded by a nuclear gene located on chromosome 10q21, is involved in the control of replication and transcription of mtDNA. To better understand the complex relationship between h-mtTFA and mitochondrial function, we assessed by western blot the levels of h-mtTFA in skeletal muscle from 7 patients affected by mitochondrial diseases (MD) caused by different mtDNA mutations. We related these results to exercise lactate production as well as to the pathologic features of the underlying myopathy. h-mtTFA levels were significantly inversely related to blood lactate and the percent of RRF, borderline to cox negative fibers. Our results indicate that hmtTFA may be involved in the cascade of events which determine functional impairment of MD. PMID- 11382205 TI - Contemporary trends and practice strategies in pediatric occupational and physical therapy. AB - This paper identifies and reflects on recent trends in pediatric occupational therapy and physical therapy practice. These trends were initially identified by the student authors for a Maternal and Child Health (MCH) leadership seminar, which was conducted as part of the postprofessional graduate program in physical therapy at the University of Washington. Trends were then reviewed and discussed among the student and faculty authors. Consensus was reached on the most important trends, which were subsequently summarized in this paper. The first part of the paper reviews the impact of these trends on current and future clinical practices in health, educational, and community-based settings. The second part of the paper offers proposed directions to meet the challenges presented by the trends in four key areas; (1) research, (2) professional education, (3) enhancing family-centered care, and (4) advocacy. PMID- 11382206 TI - Aquatic therapy in pediatrics: annotated bibliography. AB - "Aquatic therapy" refers to therapeutic intervention taking place in water. The purpose of this review is to summarize the published articles in the rehabilitation literature from 1979 through 1999 that relate to the use of aquatic therapy as an intervention for children and adolescents with neuromuscular and musculoskeletal diagnoses. Despite the trend toward evidence based practice, a paucity of literature exists related to aquatic therapy for children. Most of the available articles are case reports and other descriptions of clinical practice. The research reports are limited in design and scope, and subjects had a wide variety of ages and diagnoses. PMID- 11382207 TI - Content and construct validity of a Spanish translation of the Pediatric Evaluation of Disability Inventory for children living in Puerto Rico. AB - Although Puerto Rican children with disabilities are at high risk for poor developmental outcomes, no standardized measures are validated for use with these children. In this study, we evaluated the content and construct validity of a Spanish translation of the Pediatric Evaluation of Disability Inventory (PEDI) for use with children living in Puerto Rico. Methods included expert content review, surveys of parents, and the evaluation of 44 children with and without disabilities, who were matched for socio-economic status, age, and gender. The results demonstrated that the translated version of the PEDI is valid; yet additional research is warranted to identify socio-cultural influences on the performance and capability of typically developing Puerto Rican children. PMID- 11382208 TI - Web sites related to cerebral palsy. PMID- 11382210 TI - Using Baxter ThermaCyl blood/fluid warmer with manual pumps may cause warming cuffs to burst. PMID- 11382209 TI - Enhanced nurse call systems. AB - This Evaluation focuses on high-end computerized nurse call systems--what we call enhanced systems. These are highly flexible systems that incorporate microprocessor and communications technologies to expand the capabilities of the nurse call function. Enhanced systems, which vary in configuration from one installation to the next, typically consist of a basic system that provides standard nurse call functionality and a combination of additional enhancements that provide the added functionality the facility desires. In this study, we examine the features that distinguish enhanced nurse call systems from nonenhanced systems, focusing on their application and benefit to healthcare facilities. We evaluated seven systems to determine how well they help (1) improve patient care, as well as increase satisfaction with the care provided, and (2) improve caregiver efficiency, as well as increase satisfaction with the work environment. We found that all systems meet these objectives, but not all systems perform equally well for all implementations. Our ratings will help facilities identify those systems that offer the most effective features for their intended use. The study also includes a Technology Management Guide to help readers (1) determine whether they'll benefit from the capabilities offered by enhanced systems and (2) target a system for purchase and equip the system for optimum performance and cost-effective operation. PMID- 11382211 TI - Inadequate warnings on a GE CT/i (Octane) CT scanner may delay diagnosis. PMID- 11382212 TI - Isolated power systems. PMID- 11382213 TI - EMI from ESU impairs Kendall A-V foot pump. PMID- 11382214 TI - A graphic framework for teaching critical appraisal of randomized controlled trials. AB - Students of evidence-based medicine often try unsuccessfully to commit to memory a particular critical appraisal framework (often lengthy), or they have to depend on pocket cards and are lost without them. We have described a pedagogic aid: a flow diagram of an RCT, which has been developed over years of teaching residents. This diagram focuses on the steps in an RCT, and by drawing arrows, it highlights the biases possible at each step. This diagram serves as a framework on which the list of critical appraisal questions can be hung and is easy to remember. PMID- 11382215 TI - [Methods of measurement and biological diagnosis of the dyslipidemias in primary prevention. Text of the recommendations, January 2000. L'Agence Nationale d'Accreditation et d'Evaluation en Sante (ANAES)]. PMID- 11382216 TI - Do polyethylene glycol gels have a protective effect on the skin? PMID- 11382217 TI - Occupational post-traumatic psoriasis. PMID- 11382218 TI - Contact allergy to trometamol. PMID- 11382219 TI - The gene transfer agent of Rhodobacter capsulatus and "constitutive transduction" in prokaryotes. AB - Transduction, bacteriophage-mediated gene transfer, is thought to play an important role in the evolution of prokaryote genomes. Several gene transfer agents that resemble transducing phages have been found in diverse prokaryotes. This mini-review discusses these interesting agents of genetic exchange with a focus on the gene transfer agent (GTA) of Rhodobacter capsulatus, at present the only member of this group for which genetic information exists about the production of transducing particles. Production of GTA results from expression of genes that are similar to phage genes, yet transcription of these genes is dependent upon cellular (two-component) signaling proteins. The significance of these relationships, as well as the finding of GTA gene homologues in the bacterium Rhodopseudomonas palustris, is discussed. PMID- 11382220 TI - Microbial biosynthesis of halometabolites. AB - Halometabolites are compounds that are commonly found in nature and they are produced by many different organisms. Whereas bromometabolites can mainly be found in the marine environment, chlorometabolites are predominately produced by terrestrial organisms; iodo- and fluorocompounds are only produced infrequently. The halogen atoms are incorporated into organic compounds by enzyme-catalyzed reactions with halide ions as the halogen source. For over 40 years haloperoxidases were thought to be responsible for the incorporation of halogen atoms into organic molecules. However, haloperoxidases lack substrate specificity and regioselectivity, and the connection of haloperoxidases with the in vivo formation of halometabolites has never been demonstrated. Recently, molecular genetic investigations showed that, at least in bacteria, a different class of halogenases is involved in halometabolite formation. These halogenases were found to require FADH2, which can be produced from FAD and NADH by unspecific flavin reductases. In addition to FADH2, oxygen and halide ions (chloride and bromide) are necessary for the halogenation reaction. The FADH2-dependent halogenases show substrate specificity and regioselectivity, and their genes have been detected in many halometabolite-producing bacteria, suggesting that this type of halogenating enzymes constitutes the major source for halometabolite formation in bacteria and possibly also in other organisms. PMID- 11382221 TI - Isolation and characterization of a Chryseobacterium strain from the gut of the American cockroach, Periplaneta americana. AB - A 16S rDNA sequence cloned directly from whole-gut microbiota of the American cockroach, Periplaneta americana, indicated the presence of a member of the Bacteroides/Flavobacterium group most closely related to the genus Flavobacterium. In an attempt to confirm this finding, we isolated a yellow pigmented bacterium (strain FR2) from the hindgut of this insect. Strain FR2 was phylogentically and phenotypically most similar to species of Flavobacterium and related bacteria, namely Chryseobacterium indologenes. Fifty-four other yellow pigmented bacteria isolated during a 1-year study shared the salient phenotypic characteristics of Chryseobacterium spp., and thus were considered the same phenotype. This phenotype's abundance was related to the fiber content of the insect diet, being consistently detected only in cockroaches fed a high-fiber diet (30% crude fiber by weight). The highest population density was in the hindgut, ranging from 2 x 10(6) to 1.2 x 10(7) colony forming units ml(-1) during a 1-year period. The nature of the symbiosis between the FR2 phenotype and P. americana is discussed. PMID- 11382222 TI - Identification of amino acids in the leader peptide of Methanococcus voltae preflagellin that are important in posttranslational processing. AB - Archaeal flagellins are made initially as preproteins with short, positively charged leader peptides. Analysis of all available archaeal preflagellin sequences indicates that the -1 position is always held by a glycine while the -2 and -3 positions are almost always held by charged amino acids. To evaluate the importance of these and other amino acids in the leader peptides of archaeal flagellins for processing by a peptidase, Methanococcus voltae mutant FlaB2 preflagellin genes were generated by PCR and the proteins tested in a methanogen preflagellin peptidase assay that detects the removal of the leader peptide from preflagellin. When the -1 position was changed from glycine to other amino acids tested, no cleavage was observed by the peptidase, with the exception of a change to alanine at which poor, partial processing was observed. Amino acid substitutions at the -2 lysine position resulted in a complete loss of processing by the peptidase, while changes at the -3 lysine resulted in partial processing. A mutant preflagellin with a leader peptide shortened from 12 amino acids to 6 amino acids was not processed. When the invariant glycine residue present at position +3 was changed to a valine, no processing of this mutant preflagellin was observed. The identification of critical amino acids in FlaB2 required for proper processing suggests that a specific preflagellin peptidase may cleave archaeal flagellins by recognition of a conserved sequence of amino acids. PMID- 11382223 TI - Use of isotopic and molecular techniques to link toluene degradation in denitrifying aquifer microcosms to specific microbial populations. AB - Microcosms were inoculated with sediments from both a petroleum-hydrocarbon (PHC) contaminated aquifer and from a nearby pristine aquifer and incubated under anoxic denitrifying conditions with [methyl-13C]toluene. These microcosms served as a laboratory model system to evaluate the combination of isotope (13C-labeling of polar-lipid-derived fatty acids) and molecular techniques (16S rRNA-targeting gene probes) to identify the toluene-metabolizing population. After total depletion of toluene, the following bacterial phospholipid fatty acids (PLFA) were 13C-enriched: 16:1omega7c, 16:1omega7t, 16:0, cy17:0, and 18:1omega7c. Pure culture experiments demonstrated that these compounds were also found in PLFA profiles of PHC-degrading Azoarcus spp. (beta-Proteobacteria) and related species. The origin of the CO2 evolved in the microcosms was determined by measurements of stable carbon isotope ratios. Toluene represented 11% of the total pool of mineralized substrates in the contaminated sediment and 54% in the pristine sediment. The microbial community in the microcosm incubations was characterized by using DAPI staining and whole-cell hybridization with specific fluorescently labeled 16S rRNA-targeted oligonucleotide probes. Results revealed that 6% of the DAPI-stained cells in the contaminated sediment and 32% in the pristine sediment were PHC-degrading Azoarcus spp. In biotic control microcosms (incubated under denitrifying conditions, no toluene added), Azoarcus spp. cells remained at less than 1% of the DAPI-stained cells. The results show that isotope analysis in combination with whole-cell hybridization is a promising approach to identify and to quantify denitrifying toluene degraders within microbial communities. PMID- 11382224 TI - Molecular analysis of the cytochrome bc1-aa3 branch of the Corynebacterium glutamicum respiratory chain containing an unusual diheme cytochrome c1. AB - In this work, the genes for cytochrome aa3 oxidase and the cytochrome bc1 complex in the gram-positive soil bacterium Corynebacterium glutamicum were identified. The monocistronic ctaD gene encoded a 65-kDa protein with all features typical for subunit I of cytochrome aa3 oxidases. A ctaD deletion mutant lacked the characteristic 600 nm peak in redox difference spectra, and growth in glucose minimal medium was strongly impaired. The genes encoding subunit III of cytochrome aa3 (ctaE) and the three characteristic subunits of the cytochrome bc1 complex (qcrABC) were clustered in the order ctaE-qcrCAB. Analysis of the deduced primary structures revealed a number of unusual features: (1) cytochrome c1 (QcrC, 30 kDa) contained two Cys-X-X-Cys-His motifs for covalent heme attachment, indicating that it is a diheme c-type cytochrome; (2) the 'Rieske' iron-sulphur protein (QcrA, 45 kDa) contained three putative transmembrane helices in the N terminal region rather than only one; and (3) cytochrome b (QcrB, 60 kDa) contained, in addition to the conserved part with eight transmembrane helices, a C-terminal extension of about 120 amino acids, which presumably is located in the cytoplasm. Staining of C. glutamicum proteins for covalently bound heme indicated the presence of a single, membrane-bound c-type cytochrome with an apparent molecular mass of about 31 kDa. Since this protein was missing in a qcrCAB deletion mutant, it most likely corresponds to cytochrome c1. Similar to the deltactaD mutant, the deltaqcrCAB mutant showed strongly impaired growth in glucose minimal medium, which indicates that the bc1-aa3 pathway is the main route of respiration under these conditions. PMID- 11382225 TI - Secretion of human growth hormone by the food-grade bacterium Staphylococcus carnosus requires a propeptide irrespective of the signal peptide used. AB - Staphylococcal exoproteins can be divided into two groups. One group comprises proteins bearing only a signal peptide, the other group requires an additional propeptide for secretion. The secretion signals of the propeptide-requiring lipase from Staphylococcus hyicus (Lip) have been frequently used to produce recombinant secretory proteins in the food-grade species Staphylococcus carnosus. However, it has been unclear whether recombinant proteins can be secreted using signal peptides of staphylococcal proteins without propeptide. The human growth hormone protein (hGH) was fused to various staphylococcal secretion signals of proteins without propeptide (Seb, SceA, and SceB) and of proteins requiring a propeptide (lipase, lysostaphin, and glycerol ester hydrolase). Secretory hGH was efficiently produced by S. carnosus after fusion with any propeptide-containing secretion signal, whereas precursor proteins were retained in the cells when only a signal peptide was used. Addition of the first six amino acid residues of mature SceA to the signal peptide did also not lead to secretion of hGH. It was concluded that the properties of the mature protein domains determine whether a propeptide is required for secretion or not. The Lip propeptide could be efficiently removed from hGH after introduction of an enterokinase cleavage site between the two protein domains. PMID- 11382226 TI - Mechanisms underlying the acquisition of resistance to octanoic-acid-induced death following exposure of Saccharomyces cerevisiae to mild stress imposed by octanoic acid or ethanol. AB - Acquisition of resistance to lethal concentrations of octanoic acid was induced in cells of Saccharomyces cerevisiae grown in the presence of sublethal concentrations of this lipophilic acid or following rapid exposure (1 h) of unadapted yeast cells to mild stress imposed by the same acid. Experimental evidence indicated that the referred adaptation involved de novo protein synthesis, presumably due to the rapid induction of a plasma membrane transporter which mediates the active efflux of octanoate out of the cell. Rapid exposure of cells to mild ethanol stress also led to increased resistance to lethal concentrations of octanoic acid. This cross-resistance to octanoic-acid-induced death was below the level of resistance induced by mild octanoic acid stress and did not involve induction of the active expulsion of octanoate out of the cell. However, the rapid exposure of yeast cells to octanoic acid or ethanol led to the activation of plasma membrane H+-ATPase. The physiological role of the two stress responses examined during the present study, namely, the active efflux of octanoate specifically induced by octanoic acid and the stimulation of plasma membrane H+-ATPase activity, is discussed. PMID- 11382227 TI - Sequencing, high-level expression and phylogeny of the pyrophosphate-dependent phosphofructokinase from the thermophilic spirochete Spirochaeta thermophila. AB - The full-length gene encoding a 554-amino-acid, active pyrophosphate-dependent phosphofructokinase from Spirochaeta thermophila was cloned and sequenced using a combination of degenerate and inverse PCR, and the enzyme expressed to a high level in Escherichia coli. The recombinant enzyme, with a calculated molecular mass of 61 kDa, was purified to near homogeneity and found to be similar to the purified native enzyme for most properties examined. Phylogenetic analysis demonstrated a close relationship between the thermophilic S. thermophila phosphofructokinase and the large beta-subunits of the phosphofructokinases from Borrelia burgdorferi and Treponema pallidum. PMID- 11382228 TI - Phytochemistry in the genomics and post-genomics eras. PMID- 11382229 TI - Inhibition of polygalacturonase from Verticillium dahliae by a polygalacturonase inhibiting protein from cotton. AB - An extracellular endo-polygalacturonase (PGase) [E.C. 3.2.1.15] was isolated from 18-day-old culture filtrates of Verticillium dahliae and partially purified using gel permeation chromatography. The band responsible for PGase activity was electrophoretically characterized as having a molecular mass of approximately 29 500 and an isoelectric point of 5.4. Kinetic studies indicate a Km of 3.3 mg ml( 1) and Vmax of 0.85 micromol reducing units min(-1) ml(-1) with polygalacturonic acid as substrate. Polygalacturonase inhibitor protein (PGIP) in cotton seedlings was induced by 5 mM salicylic acid and immunochemical analysis indicated high levels in the hypocotyl tissues. PGIP was purified from roots and stems using affinity chromatography with endo-PGase from Aspergillus niger as an immobilised ligand. The purified PGIP contained monomeric and dimeric molecules with molecular masses of 34 and 66 kDa respectively. Purified cotton PGIP inhibited endo-polygalacturonase from A. niger in a non-competitive or mixed manner with an inhibition constant. K(I) of 15 nM. The isolated V. dahliae PGase was, however, inhibited in a positive cooperative manner, indicative of allosteric interactions between the enzyme and the inhibitor protein. In addition to reducing the reaction rate, decreased substrate affinity may contribute to the accumulation of elicitor-active oligouronides. PMID- 11382230 TI - Cell-wall proteins from Sitka spruce xylem are selectively insolubilised during formation of dehydrogenation polymers of coniferyl alcohol. AB - Extracts from the lignifying xylem of Sitka spruce that were enriched in cell wall-associated glycoproteins contained peroxidase and oxidase activity and readily formed lignin-like water-insoluble dehydrogenation polymers (DHPs) from coniferyl alcohol (CA) when supplied with H2O2. During the formation of DHPs, the abundance of a number of polypeptides in the extracts was diminished. However, these polypeptides were also diminished in control reactions that contained H2O2 but lacked CA. Polypeptides could be recovered from the DHPs by heating in SDS PAGE sample buffer but no insolubilised polypeptides could be recovered from the + H2O2 reactions. Although most of the DHP-bound polypeptides were easily removed by pre-washing the DHPs, two polypeptides at 125 and 52 kDa remained tightly bound to the DHPs. The abundance of the two DHP-bound polypeptides mirrored the diminution of 120 and 46 kDa polypeptides in the extracts. The N-terminal protein sequences of the 125 and 52 kDa DHP-bound polypeptides were essentially identical to the sequences obtained from the 120 and 46 kDa polypeptides from the extracts, which confirmed that the DHP-bound polypeptides were derived from these soluble polypeptides. The 125-kDa DHP-bound polypeptide yielded an N-terminal protein sequence that was identical to a laccase-type oxidase previously identified in similar extracts from lignifying Sitka xylem. The N-terminal protein sequence of the 46-kDa polypeptide was homologous with a subset of plant peroxidases. The DHPs had tightly bound peroxidase and oxidase activity, which suggested that these polypeptides were active in their insolubilised state. The mechanism and selectivity of insolubilisation of these enzymes is discussed. PMID- 11382231 TI - Biosynthesis of diaporthin and orthosporin by Aspergillus ochraceus. AB - Diaporthin and orthosporin were characterised from the fungus Aspergillus ochraceus D2306. Diaporthin was identified by high-resolution electron impact mass spectrometry and 1H and 13C NMR spectroscopy, from which new spectroscopic assignments were made. Orthosporin was also identified by mass spectrometry and both fungal metabolites are reported for the first time as co-metabolites and also as products of A. ochraceus. The methylation inhibitor ethionine affected production of both diaporthin and orthosporin in spite of no obvious methylation step in the biosynthesis of orthosporin, implying that extracellular orthosporin may arise by de-O-methylation of diaporthin. The biosynthetic origin of diaporthin was demonstrated by incorporation of [1-14C]acetate and [methyl 14C]methionine administered in early idiophase. PMID- 11382232 TI - Accumulation of 6-deoxocathasterone and 6-deoxocastasterone in Arabidopsis, pea and tomato is suggestive of common rate-limiting steps in brassinosteroid biosynthesis. AB - To gain a better understanding of brassinosteroid biosynthesis, the levels of brassinosteroids and sterols related to brassinolide biosynthesis in Arabidopsis, pea, and tomato plants were quantified by gas chromatography-selected ion monitoring. In these plants, the late C-6 oxidation pathway was found to be the predominant pathway in the synthesis of castasterone. Furthermore, all these plant species had similar BR profiles, suggesting the presence of common biosynthetic control mechanisms. The especially high levels of 6 deoxocathasterone and 6-deoxocastasterone may indicate that their respective conversions to 6-deoxoteasterone and castasterone are regulated in planta and hence are important rate-limiting steps in brassinosteroid biosynthesis. Other possible rate-limiting reactions, including the conversion of campestanol to 6 deoxocathasteonre. are also discussed. Tomato differs from Arabidopsis and pea in that tomato contains 28-norcastasterone as a biologically active brassinosteroid, and that its putative precursors, cholesterol and its relatives are the major sterols. PMID- 11382233 TI - Metabolism of indole-3-acetic acid by orange (Citrus sinensis) flavedo tissue during fruit development. AB - [5-3H, 1'-14C, 13C6, 12C] Indole-3-acetic acid (IAA), was applied to the flavedo (epicarp) of intact orange fruits at different stages of development. After incubation in the dark, at 25 degrees C, the tissue was extracted with MeOH and the partially purified extracts were analyzed by reversed phase HPLC-RC. Six major metabolite peaks were detected and subsequently analyzed by combined HPLC frit-FAB MS. The metabolite peak 6 contained oxindole-3-acetic acid (OxIAA), indole-3-acetyl-N-aspartic acid (IAAsp) and also indole-3-acetyl-N-glutamic acid (IAGlu). The nature of metabolite 5 remains unknown. Metabolites 3 and 4 were diastereomers of oxindole-3-acetyl-N-aspartic acid (OxIAAsp). Metabolite 2 was identified as dioxindole-3-acetic acid and metabolite 1 as a DiOx-IAA linked in position three to a hexose, which is suggested to be 3-(-O-beta-glucosyl) dioxindole-3-acetic acid (DiOxIAGlc). Identification work as well as feeding experiments with the [5-3H]IAA labeled metabolites suggest that IAA is metabolized in flavedo tissue mainly through two pathways, namely IAA-OxIAA DiOxIAA-DiOxIAGlc and IAA-IAAsp-OxIAAsp. The flavedo of citrus fruit has a high capacity for IAA catabolism until the beginning of fruit senescence, with the major route having DiOxIAGlc as end product. This capacity is operative even at high IAA concentrations and is accelerated by pretreatment with the synthetic auxins 2,4-D, NAA and the gibberellin GA3. PMID- 11382234 TI - The uptake and speciation of various Al species in the Brassica rapa pekinensis. AB - An investigation was carried out on the uptake and speciation of Al species in Al tolerant Chinese cabbage (Brassica rapa L. ssp. pekinensis). Plants were exposed to 10 microg cm(-3) of Al in the chemical forms of Al3+, Al-citrate and Al-malate in a time span from 1 up to 24 h. In each experiment the nutrient solution and stem sap were analysed by a combination of FPLC ICP AES and ES MS MS techniques. Speciation analysis enabled determination of particular chemical forms of Al present in the nutrient solution or in stem sap. The results indicate that Al3+ added to the nutrient solution remained as Al3+ in the solution during the experiments, but in the roots transformation to Al-malate occurred. Al was transported from roots to the upper parts of the plant as Al-malate (70%) and Al3+ (30%). Al-citrate or Al-malate added to the nutrient solution were transferred to the upper parts of the plant without transformation of their chemical forms. PMID- 11382235 TI - Biotransformation of (R)-(+)- and (S)-(-)-limonene by fungi and the use of solid phase microextraction for screening. AB - The biotransformation of (R)-(-)- and (S)-(-)-limonene by fungi was investigated. More than 60 fungal cultures were screened for their ability to bioconvert the substrate, using solid phase microextraction as the monitoring technique. After screening, the best fungal strains were selected for further study and were grown as sporulated surface cultures in conical flasks and as submerged liquid cultures. It was found that (+)- and (-)-limonene were converted by Penicillium digitatum to alpha-terpineol (main metabolite), cis- and trans-p-menth-2-en-1-ol, neodihydrocarveol and limonene oxide (minor metabolites) using liquid cultures. The bioconversion of (R)-(-)- and (S)-(-)-limonene by Corwespora cassiicola yielded (1S,2S,4R)- and (1R,2R,4S)-limonene-1,2-diol respectively. The bioconversions by liquid cultures were also monitored by solid phase microextraction as a function of time. The optimum conversion of limonene to alpha-terpineol by Penicillium digitatum was obtained after 8 hours (yield up to 100%). Since an important pH-decrease was noticed in some liquid broths, the stability of limonene under acidic conditions was investigated. No acid catalysed conversion products were recovered after 8 days from control flasks at pH 3.5 containing limonene. PMID- 11382236 TI - Comprehensive chemical profiling of gramineous plant root exudates using high resolution NMR and MS. AB - Root exudates released into soil have important functions in mobilizing metal micronutrients and for causing selective enrichment of plant beneficial soil micro-organisms that colonize the rhizosphere. Analysis of plant root exudates typically has involved chromatographic methods that rely on a priori knowledge of which compounds might be present. In the research reported here, the combination of multinuclear and 2-D NMR with GC-MS and high-resolution MS provided de novo identification of a number of components directly in crude root exudates of different plant types. This approach was applied to examine the role of exudate metal ion ligands (MIL) in the acquisition of Cd and transition metals by barley and wheat. The exudation of mugineic acids and malate was enhanced by Fe deficiency. which in turn led to an increase in the tissue content of Cu, Mn, and Zn. The presence of elevated Cd maintained at a free activity pCd of 8.8 (10( 8.8) M), resulted in reduced phytosiderophore production by Fe deficient plants. The buffer morpholinoethane sulfonate (MES), which is commonly used in chelator buffering nutrient solutions, was detected in the root exudate mixture, suggesting uptake and re-secretion of this compound by the roots. The ability to detect this compound in complex mixtures containing organic acids, amino acids, and other substances suggests that the analytical methods used here provide an unbiased method for simultaneous detection of all major components contained in root exudates. PMID- 11382237 TI - Phytoalexins from Pinus strobus bark infected with pinewood nematode, bursaphelenchus xylophilus. AB - From the bark of Pinus strobus infected with pinewood nematode, Bursaphelenchus xylophilus, a stilbenoid 3-O-methyldihydropinosylvin and a flavanone (2S) pinocembrin were isolated as active principles of inducibly produced antifungal compounds. The structures of the compounds were determined on the basis of spectroscopic analyses. Close investigations of spectroscopic analyses led to the complete assignment of 13C NMR and 1H NMR chemical shifts for the former compound and to the determination of the stereochemistry of the latter compound from P. strobus to be 2S. Furthermore, the distribution and time course accumulation of the identified compounds were investigated. The former compound was demonstrated to be nematocidal and the concentrations of the compound in the inoculated branches were sufficient enough to inactivate the pinewood nematode within one week after inoculation. PMID- 11382238 TI - Synergism between the potato glycoalkaloids alpha-chaconine and alpha-solanine in inhibition of snail feeding. AB - Snails (Helix aspersa L.) were fed filter paper treated with the potato glycoalkaloids, alpha-solanine and alpha-chaconine, singly or together. In pure form, both glycoalkaloids deterred feeding, with chaconine being the more active compound. In combination, authentic solanine and chaconine interacted synergistically in their inhibition of feeding. The antifeedant activities of methanolic extracts of tuber peel of the potato varieties Majestic and Sharpe's Express presented via filter paper discs did not differ significantly from those of authentic glycoalkaloid solutions of comparable concentration and ratio. In contrast, feeding inhibition by diluted tuber peel extracts of the variety Homeguard was greater than that elicited by comparable authentic glycoalkaloid solutions suggesting additional inhibitory compound(s) in the peel of this variety. Comparison of data from peel extracts of all three potato varieties and authentic glycoalkaloids indicated that the level of feeding inhibition by the extracts was, at least in part, a consequence of a synergism between solanine and chaconine. PMID- 11382239 TI - A chemotaxonomic investigation of volatile constituents in Stachys subsect. Swainsonianeae (Labiatae). AB - The composition of the previously unknown essential oils of Stachys subsect. Swainsonianeae (Labiatae) are investigated by GC-MS analyses. The group includes the taxa Stachys ionica, St. spruneri, St. swainsonii ssp. swainsonii, St. swainsonii ssp. argolica, St. swainsonii ssp. melangavica and St. swainsonii ssp. scyronica, all endemic to Greece. Eighteen populations were analyzed, and more than two hundred components were identified. Statistical analyses of oil components separated St. ionica, due to its high amount of (E)-nerolidol, high amount of alpha-cadinol and low amount of (+)-(E)-caryophyllene. The rest of the taxa forms a coherent group where (+)-(E)-caryophyllene, delta-cadinene and partly beta-elemene, (+)-caryophyllene oxide and (+)-limonene predominate but each of them never exceeds 20% of the total. Essential oil composition reflects current taxonomy and phytogeography of the group with a somewhat isolated position for St. ionica. Similarities in morphology and essential oil composition of the St. swainsonii group and St. spruneri may be the result of extensive gene flow in the past. Evidence from volatile oil compounds that may attribute to St. swainsonii ssp. melangavica a hybrid origin from crosses between St. spruneri and St. swainsonii ssp. swainsonii/scyronica group is scarce. PMID- 11382240 TI - Essential oils from Azorean Laurus azorica. AB - The essential oils isolated from leaves of ten and from unripe berries of eight populations of Laurus azorica (Seub.) Franco, collected on five islands of the Azorean archipelago, were analysed by GC and GC-MS. All oil samples were dominated by their monoterpene fraction (60-94%), alpha-pinene (15-37%) and 1,8 cineole (12-31%) being the main components of the leaf oils, while trans-beta ocimene (27-45%) and alpha-pinene (12-22%) were the main components of the oils from the berries. The sesquiterpene fractions of the oils ranged from 3 to 17% and the main components were beta-caryophyllene (traces-8%) and beta-elemene (traces-3%) both in the leaf and berry oils. Some phenylpropanoid components were also present, in total amounting to 17%, trans-cinnamyl acetate (215% of the leaf oils) being the main component of this fraction. Cluster analysis of the enantiomeric composition of alpha- and beta-pinene in the oils from the leaves clearly showed two groups, one constituted by the two populations growing on the island S. Jorge, and the other constituted by the remaining populations. PMID- 11382241 TI - Cyclolinopeptides F-I, cyclic peptides from linseed. AB - Four cyclic peptides, cyclolinopeptides F-I, were isolated from seeds of Linum usitatissimum. Their structures were elucidated by extensive 2D NMR spectroscopic methods and by chemical degradation. Further, their immunosuppressive activity is examined. PMID- 11382242 TI - Jesterone and hydroxy-jesterone antioomycete cyclohexenone epoxides from the endophytic fungus Pestalotiopsis jesteri. AB - Jesterone and hydroxy-jesterone are novel highly functionalized cyclohexenone epoxides isolated from a newly described endophytic fungal species- Pestalotiopsis jesteri. They were purified from cultures of the fungus by bioassay guided fractionation using Pythium ultimum as the indicator organism. Jesterone, in particular, displays selective antimycotic activity against the oomycetous fungi which are some of the most plant pathogenic of all disease causing fungi. The possible importance of these cyclohexenones to the biology of the endophytic fungus-host plant relationship is also discussed. PMID- 11382243 TI - Sesquiterpene lactones and a myoinositol from glandular trichomes of Viguiera quinqueremis (Heliantheae; Asteraceae). AB - The extract of the floral parts of Viguiera quinqueremis afforded, in addition to known compounds, six new sesquiterpene lactones as well as a new myoinositol derivative. All compounds were detected in glandular trichomes which were collected micromechanically from the anther appendages and were analyzed by HPLC. Structure identification was performed by 1H NMR measurements including LC NMR and LC MS experiments. PMID- 11382244 TI - Benzopyran derivatives from Mallotus apelta. AB - From the leaves of Mallotus apelta, seven benzopyran compounds were obtained and their structures were determined using spectroscopic methods. One showed moderate antibiotic activity against Micrococcus lutens. PMID- 11382245 TI - Nemorosone, the major constituent of floral resins of Clusia rosea. AB - Nemorosone, the major constituent of the floral resin of Clusia rosea was isolated after exhaustive chromatography. This compound was fully characterized as it is in the nature, without methylation as reported before. A keto-enol equilibrium was observed and both isomers were totally characterized by NMR spectroscopic techniques. The previously announced structure for methylnemorosone was corrected on the basis of application of chemical methylation, high field 2D NMR techniques and NOE difference spectroscopy experiments on the natural product. Our studies concluded that an interchange occurred in the assignment of the benzoyl moiety position with an isoprenyl group in that structure. PMID- 11382246 TI - Minor constituents of Spigelia anthelmia and their cardiac activities. AB - A more detailed phytochemical analysis of extracts of the aerial parts of Spigelia anthelmia L. (Loganiaceae) yielded 20 structurally related new compounds besides spiganthine and ryanodine. Structure elucidation was achieved mainly by spectroscopic methods. The compounds were tested on their cardiac and on their insect antifeedant activities. PMID- 11382247 TI - Flavonoid glucuronides from Helicteres isora. AB - Five flavonoid glucuronides were obtained from the fruit of Helicteres isora, three of which were previously unknown compounds: isoscutellarein 4'-methyl ether 8-O-beta-D-glucuronide 6"-n-butyl ester. isoscutellarein 4'-methyl ether 8-O-beta D-glucuronide 2", 4"-disulfate and isoscutellarein 8-O-beta-D-glucuronide 2",4" disulfate. The structures were determined on the basis of spectroscopy and hydrolysis experiments. PMID- 11382248 TI - Rapid detection of chlorinated bisbibenzyls in Bazzania trilobata using MALDI-TOF mass spectrometry. AB - Chlorinated bisbibenzyls of the bazzanin type are detected in crude bryophyte plant extracts of Bazzania trilobata from different locations using MALDI-TOF mass spectrometry. These results show that these chlorinated compounds are not artefacts of an incidental occurrence or of the sample preparation but are genuine and produced by the liverwort or an endosymbiotic metabolism. Further experiments were performed concerning the in vitro chlorination of the halogen free basic unit isoplagiochin C. PMID- 11382249 TI - Pacifigorgianes and tamariscene as constituents of Frullania tamarisci and Valeriana officinalis. AB - The sesquiterpenoid constituents of the essential oils from the liverworts Frullania tamarisci, Frullania fragilifolia and of the angiosperm Valeriana officinalis were investigated. Tamariscene, a compound with a new sesquiterpene skeleton, valerena-4,7(11)-diene and five new pacifigorgiadienes, namely pacifigorgia-1,10-diene, pacifigorgia-1(6),10-diene, pacifigorgia-1(9),10-diene, pacifigorgia-2,10-diene, and pacifigorgia-2(10),11-diene were isolated and identified. Structure elucidation was carried out by NMR spectroscopy and chemical correlations to establish absolute configurations. Compounds present in both the essential oils of the Frullania species and Valeriana officinalis were enantiomeric to each other. A plausible biogenetic relationship between the pacifigorgiane, valerenane and tamariscane skeletons is postulated. Pacifigorgia 6,11-diene, not yet detected in nature, was generated by dehydration and rearrangement of natural (-)-tamariscol. PMID- 11382250 TI - Outcome evaluation in patients with elbow pathology: issues in instrument development and evaluation. AB - Measurement of outcome after treatment of elbow pathology has tended to rely on nonstandardized scales and isolated impairment measures, particularly range of motion. This study evaluated the reliability and validity of patient questionnaires with patients with elbow pathology. Patients with a variety of complaints affecting the elbow (n = 70) completed four questionnaires-the American Shoulder and Elbow Surgeons Elbow Form; the Patient-rated Elbow Evaluation; the Disabilities of Arm, Shoulder and Hand questionnaire; and Short Form 36--during clinic attendance and returned repeat questionnaires by mail (n = 50). The test-retest reliability was determined to be acceptable for all four instruments (16 of 19 ICCs for subscales > 0.78; all ICCs for total score s > 0.90). Four constructs on the relationship expected between outcome measures, determined prior to data collection, were observed as anticipated, supporting the validity of outcome questionnaires. This study supports the use of these instruments to evaluate outcome in patients with elbow pathology. PMID- 11382251 TI - An upper body musculoskeletal assessment instrument for patients with work related musculoskeletal disorders: a pilot study. AB - OBJECTIVES: To examine the reliability and validity of a new outcome measure, the Upper Body Musculoskeletal Assessment (UBMA). DESIGN: Twenty subjects physician diagnosed as having work-related musculoskeletal disorders (WRMD) and ten healthy subjects were assessed using the UBMA on three separate occasions. All subjects with WRMD attributed their injury to equipment use on their job. RESULTS: The WRMD group had significantly higher UBMA scores on the side of equipment use than on the other side (p <0.01), whereas the healthy group had similar scores on both sides (p> 0.05). UBMA scores for the WRMD group were significantly greater on both sides of the body than scores for the healthy group (p<0.01). Only one test occasion was required to produce excellent reliability coefficients (ICCs>0.88). Although group reliability was excellent, changes of 24% for patients with WRMD and 44% for healthy subjects would be required for confidence that UBMA scores for individual patients on the side of equipment use had changed from baseline. CONCLUSIONS: Although testing on one occasion produced reliable UBMA scores, healthy subjects could be distinguished from patients with WRMD, and the side of equipment use could be distinguished from the other side in patients with WRMD, prediction of individual UBMA scores was poor. In its present form, the UBMA is useful for making decisions about groups but not about individual patients. Modifications of the current UBMA are required to reduce measurement error. PMID- 11382252 TI - The Injured Workers Survey: development and clinical use of a psychosocial screening tool for patients with hand injuries. AB - Hand therapists often recognize that their patients are experiencing psychosocial distress following traumatic hand injury but may lack the time and training to determine whether referral for psychological assessment and treatment is warranted. The Injured Workers Survey (IWS) is a brief, effective self-report tool that screens for psychosocial distress, in particular for characteristic symptoms of post-traumatic stress disorder (PTSD). It also collects clinically relevant information about other psychosocial issues that have a major effect on treatment outcomes. A recent research study has validated the IWS as a screen for PTSD and supports its routine use in the initial assessment of all patients with traumatic hand injuries. To the extent that psychosocial distress interferes with recovery, its early identification and treatment can optimize overall treatment outcomes and facilitate successful in return to work. PMID- 11382253 TI - Measuring the whole or the parts? Validity, reliability, and responsiveness of the Disabilities of the Arm, Shoulder and Hand outcome measure in different regions of the upper extremity. AB - The Disabilities of the Arm, Shoulder and Hand (DASH) outcome measure was developed to evaluate disability and symptoms in single or multiple disorders of the upper limb at one point or at many points in time. PURPOSE: The purpose of this study was to evaluate the reliability, validity, and responsiveness of the DASH in a group of diverse patients and to compare the results with those obtained with joint-specific measures. METHODS: Two hundred patients with either wrist/hand or shoulder problems were evaluated by use of questionnaires before treatment, and 172 (86%) were re-evaluated 12 weeks after treatment. Eighty-six patients also completed a test-retest questionnaire three to five days after the initial (baseline) evaluation. The questionnaire package included the DASH, the Brigham (carpal tunnel) questionnaire, the SPADI (Shoulder Pain and Disability Index), and other markers of pain and function. Correlations or t-tests between the DASH and the other measures were used to assess construct validity. Test retest reliability was assessed using the intraclass correlation coefficient and other summary statistics. Responsiveness was described using standardized response means, receiver operating characteristics curves, and correlations between change in DASH score and change in scores of other measures. Standard response means were used to compare DASH responsiveness with that of the Brigham questionnaire and the SPADI in each region. RESULTS: The DASH was found to correlate with other measures (r > 0.69) and to discriminate well, for example, between patients who were working and those who were not (p<0.0001). Test-retest reliability (ICC = 0.96) exceeded guidelines. The responsiveness of the DASH (to self-rated or expected change) was comparable with or better than that of the joint-specific measures in the whole group and in each region. CONCLUSIONS: Evidence was provided of the validity, test-retest reliability, and responsiveness of the DASH. This study also demonstrated that the DASH had validity and responsiveness in both proximal and distal disorders, confirming its usefulness across the whole extremity. PMID- 11382254 TI - Validity of health status measures in patients with ulnar wrist disorders. AB - PURPOSE: To assess measurement properties and construct validity of health status measures. METHOD: Forty-three patients with surgically managed ulnocarpal impingement completed a self-report mail survey, including regional (Disabilities of the Arm, Shoulder and Hand [DASH] questionnaire), disease-specific (Brigham Functional Scale), and generic (Short Form 36 [SF-36] Acute Health Survey) health status measures and questions on condition severity and work status. STATISTICAL ANALYSIS: Scores were transformed and frequency distributions constructed to compare the distribution of responses to the measures. Correlation analysis and analysis of variance were applied to assess construct validity. RESULTS: The DASH and Brigham questionnaires had similar distributions of scores, with a slightly greater spread of responses and no ceiling effect on the DASH. Patients appeared slightly less healthy on the basis of the SF-36 scores, which reflected in part the effect of comorbidities. Both the DASH and the Brigham discriminated across levels of severity and work status; the DASH also discriminated on the basis of type of surgery. The SF-36 was able to discriminate some constructs but not as well as the regional and disease-specific measures. CONCLUSION: This study provides evidence of construct validity for the DASH and Brigham questionnaires in patients with ulnar wrist problems in the late post-operative period. PMID- 11382255 TI - Distal radius fracture: a prospective outcome study of 275 patients. PMID- 11382256 TI - Outcome assessment in hand surgery and hand therapy: an update. PMID- 11382257 TI - A review of self-report scales for the assessment of functional limitation and disability of the shoulder. AB - A shoulder disorder affects a patient's overall quality of life. Self-report outcome measures can be used to assess the effects of shoulder dysfunction in terms of patients' functional limitations and disabilities. Selection of a shoulder-specific scale from the numerous scales available can be overwhelming. The purpose of this study was to review the self-report scales in current use for assessing the shoulder joint. A MEDLINE literature search was performed to locate scales appropriate to the assessment of function and disability of the shoulder. Only scales with established measurement properties were selected for review. Eleven scales were reviewed. Methodological and practical criteria were established to evaluate each scale. All scales met the minimal criteria, but not all scales were equal with regard to the criteria established for analysis. PMID- 11382258 TI - Outcome measures for evaluation of scar: a literature review. AB - The number of interventions and the related costs of management of scar are increasing. Despite increasing emphasis on evidence-based practice, there is little comparative data on which to base treatment selection. The ability of a therapist to evaluate the natural progression of a scar or treatment efficacy may be hampered by the nature of the particular scar, the lack of conformity in descriptions of scar, and the lack of reliable standardized outcome measurements. This literature review seeks to clarify the constructs measured for scar and present the measures currently available for use. PMID- 11382259 TI - Pain outcome measures. AB - Pain is a complex, multi-dimensional experience that is usually associated with local tissue damage or may be referred from a distant site. Classically, pain is viewed as having sensory, affective, and cognitive components. To assess pain, however, the clinician or the researcher must use the most appropriate measure for the given situation. Many pain outcome measures are currently in use. Some of these are simple uni-dimensional scales, whereas others adopt a more multi dimensional approach. Pain is rarely measured in isolation but must instead be put into the context of the whole person and that person's functional ability. All these factors should be borne in mind in the evaluation of pain in the hand. This paper discusses pain measurement instruments and makes recommendations for assessment of pain in the hand. PMID- 11382260 TI - The Prosthetic Upper Extremity Functional Index: development and reliability testing of a new functional status questionnaire for children who use upper extremity prostheses. AB - The Prosthetic Upper Extremity Functional Index (PUFI) was developed by the authors' clinical research group to evaluate the extent to which a child actually uses a prosthetic limb for daily activities, the comparative ease of task performance with and without the prosthesis, and its perceived usefulness. The PUFI's test-retest and interrater reliability were evaluated with 24 children. Intraclass coefficients (ICCs) were calculated for each of four subscales of the PUFI--specifically, method of performance, ease of prosthetic use, usefulness of the prosthesis, and ease of performance without the prosthesis. The ICCs were greater than 0.65, indicating good test-retest reliability for the older-child respondents (n = 10) and fair to good reliability (ICCs, 0.40 to 0.84) for the parent respondents overall (n= 21). Interrater (child-parent) reliability was lower, with ICCs from 0.30 to 0.77. This finding was not unexpected, since a child and parent may rate in the context of different functional environments. The prosthesis was used 53% of the time by older children and more than 75% of the time by younger children. The results provide evidence that the PUFI has good test-retest reliability overall as a measure of a child's ability to perform upper extremity activities with a prosthesis. PMID- 11382261 TI - Teaching psychosomatic (biopsychosocial) medicine in United States medical schools: survey findings. AB - OBJECTIVE: A survey of US medical schools regarding the incorporation of psychosomatic (biopsychosocial) medicine topics into medical school curriculum was conducted. The perceived importance and success of this curriculum, barriers to teaching psychosomatic medicine, and curricular needs were also assessed. METHODS: From August 1997 to August 1999, representatives of US medical schools were contacted to complete a survey instrument either by telephone interview or by written questionnaire. RESULTS: Survey responses were received from 54 of the 118 US medical schools contacted (46%). Responses were obtained from representatives of both public (57%) and private (43%) institutions. Only 20% of respondents indicated that their schools used the term "psychosomatic medicine"; the terms "behavioral medicine" (63%) and "biopsychosocial medicine" (41%) were used more frequently. Coverage of various health habits (eg, substance use and exercise) ranged from 52% to 96%. The conceptualization and/or measurement of psychosocial factors (eg, stress and social support) was taught by 80% to 93% of schools. Teaching about the role of psychosocial factors in specific disease states or syndromes ranged from 33% (renal disease) to 83% (cardiovascular disease). Coverage of treatment-related issues ranged from 44% (relaxation/biofeedback) to 98% (doctor-patient communication). Topics in psychosomatic medicine were estimated to comprise approximately 10% (median response) of the medical school curriculum. On a scale of 1 (lowest) to 10 (highest), ratings of the relative importance of this curriculum averaged 7 (SD = 2.5; range = 2-10). Student response to the curriculum varied from positive to mixed to negative. Perceived barriers to teaching psychosomatic medicine included limited resources (eg, time, money, and faculty), student and faculty resistance, and a lack of continuity among courses. Sixty-three percent of respondents expressed an interest in receiving information about further incorporation of topics in psychosomatic medicine into their school's curriculum. CONCLUSIONS: Results of this survey reveal variable coverage of specific psychosomatic medicine topics in the medical school curriculum and differential use of nomenclature to refer to this field. There is a need for further curricular development in psychosomatic medicine in US medical schools. PMID- 11382262 TI - Intimate partner violence and health: self-assessed health, chronic health, and somatic symptoms among Mexican American women. AB - OBJECTIVE: In medical settings intimate partner violence (IPV) has been linked to a variety of health problems. However, few population-based studies have assessed the health of abused women, particularly women from low socioeconomic groups such as Mexican Americans. This study examined the association between recent physical or sexual IPV and self-rated health, chronic health conditions, and somatic symptoms among Mexican American women. METHODS: Participants were women (N = 1155) with current male partners enrolled in a household survey of 3012 Mexican origin adults, ages 18 to 59 years, living in urban, town, and rural areas of Fresno County, California. Crude and adjusted odds ratios (ORs) were calculated for four self-assessed health measures, seven chronic diseases, and 32 somatic symptoms. RESULTS: In multivariate analyses, women reporting previous-year physical or sexual IPV were more likely to report 1) fair/poor overall health (OR, 1.9; confidence interval [CI], 1.0-3.7), physical health (OR, 2.1; CI, 1.2 3.9), and mental health (OR, 3.4; CI, 1.9-6.1), as well as worse comparative health (OR, 4.4; CI, 2.3-8.3); 2) a history of heart problems (OR, 17.0; CI, 4.3 66.7); 3) persistent health problems (OR, 3.3; CI, 1.5-7.0); and 4) numerous somatic symptoms. CONCLUSIONS: Physical or sexual IPV was associated with poorer self-assessed health and many health symptoms among this culturally distinctive Mexican American population. PMID- 11382263 TI - Are women with premenstrual dysphoric disorder prone to osteoporosis? AB - OBJECTIVE: The objective of this study was to examine whether bone mineral density (BMD) is reduced in women with premenstrual dysphoric disorder (PMDD). METHODS: Thirty-eight healthy women participated: 20 with prospectively confirmed PMDD and 18 without PMDD. Bone mass was measured using dual-energy x-ray absorptiometry at three sites: lumbar spine anteroposterior, lumber spine lateral, and femoral neck. Results from the PMDD and control groups were compared with each other and with age- and sex-matched normative data. RESULTS: The BMD of both groups was as expected for their age and sex, and groups did not differ in BMD or Z scores for any of the bone sites studied. CONCLUSIONS: If women with PMDD are at an increased risk of developing osteoporosis, this risk is not manifested in their BMD. PMID- 11382264 TI - Cerebral activation in patients with irritable bowel syndrome and control subjects during rectosigmoid stimulation. AB - OBJECTIVE: Patients with irritable bowel syndrome (IBS) show evidence of altered perceptual responses to visceral stimuli, consistent with altered processing of visceral afferent information by the brain. In the current study, brain responses to anticipated and delivered rectal balloon distension were assessed. METHODS: Changes in regional cerebral blood flow were measured using H2(15)O-water positron emission tomography in 12 nonconstipated IBS patients and 12 healthy control subjects. Regional cerebral blood flow responses to moderate rectal distension (45 mm Hg) and anticipated but undelivered distension were assessed before and after a series of repetitive noxious (60-mm Hg) sigmoid distensions. RESULTS: Brain regions activated by actual and simulated distensions were similar in both groups. Compared with control subjects, patients with IBS showed lateralized activation of right prefrontal cortex; reduced activation of perigenual cortex, temporal lobe, and brain stem; but enhanced activation of rostral anterior cingulate and posterior cingulate cortices. CONCLUSIONS: IBS patients show altered brain responses to rectal stimuli, regardless of whether these stimuli are actually delivered or simply anticipated. These alterations are consistent with reported alterations in autonomic and perceptual responses and may be related to altered central noradrenergic modulation. PMID- 11382265 TI - Timing and specificity of the cognitive changes induced by interleukin-2 and interferon-alpha treatments in cancer patients. AB - OBJECTIVE: Neuropsychological changes develop in patients treated by cytokine immunotherapy with interleukin-2 (IL-2) and interferon-alpha (IFN-alpha). However, the time course of appearance of these effects remains unclear, and their precise nature is still incompletely characterized. The objective of this study was to assess and characterize the early cognitive changes induced by IL-2 and IFN-alpha in cancer patients at the end of the first week of treatment and to investigate the subsequent evolution of these changes. METHODS: The study was conducted in 47 cancer patients who received subcutaneous IL-2, administered alone (N = 17) or with IFN-alpha (N = 7), or IFN-alpha alone, administered subcutaneously at low doses (N = 7) or intravenously at high doses (N = 16). An automated battery of neuropsychological tests (Cambridge Neuropsychological Test Automated Battery) was used to measure reaction time, spatial working memory, and planning tasks. Cognitive tests were performed before treatment (day 1) and after 5 days (day 5) and 1 month of treatment. RESULTS: On day 5, patients treated with IL-2 alone had impaired spatial working memory and lower accuracy of planning abilities. In contrast, patients treated with IFN-alpha did not show any impairment in performance accuracy in these tasks but showed longer latencies in the test of reaction time. Most of these early alterations persisted at the end of the first month of treatment without any obvious sign of worsening. CONCLUSIONS: These findings suggest the existence of early differential neuropsychological changes in patients treated with IL-2 and IFN-alpha. PMID- 11382267 TI - Predictors of posttraumatic stress among victims of motor vehicle accidents. AB - OBJECTIVE: This study identified factors that predict individual vulnerability to psychological trauma by examining the relationships among situation and person variables and symptoms of posttraumatic stress disorder (PTSD) 1, 6, and 12 months after a serious motor vehicle accident (MVA). METHODS: Background characteristics, exposure variables (ie, injury severity and accident characteristics), and psychosocial variables (ie, perceived loss of control, social support, and coping) were used to predict symptoms of PTSD and recovery in 115 injured MVA victims. All participants were injured during the MVA and provided data prospectively over the course of a year after their accidents. RESULTS: Along with background and exposure variables, use of wishful thinking coping distinguished between victims with and without symptoms of PTSD. CONCLUSIONS: Psychosocial variables such as wishful thinking coping can be used to identify MVA victims who are at risk of developing chronic posttraumatic stress and warrant further investigation. PMID- 11382266 TI - Psychogenic lowering of urinary cortisol levels linked to increased emotional numbing and a shame-depressive syndrome in combat-related posttraumatic stress disorder. AB - OBJECTIVE: The purpose of the study was to search for the intrapsychic correlates of individual differences in cortisol levels in male Vietnam combat veterans with posttraumatic stress disorder. METHODS: The study involved measurement of urinary cortisol levels and clinical assessment with a broad profile of psychometric tests during a single 48-hour period in 30 inpatients. RESULTS: The main finding by both correlation and t test analyses was a significant inverse relationship between urinary cortisol levels and a symptom complex composed of two closely interrelated clinical subgroupings, "disengagement" (principally involving emotional numbing) and "shame-laden depression." CONCLUSIONS: The findings support the concept that cortisol levels reflect the ongoing balance between the undifferentiated emotional arousal state of engagement (associated with higher cortisol levels) and opposing antiarousal disengagement defense mechanisms (associated with lower cortisol levels). It appears that the low cortisol levels often seen in patients with posttraumatic stress disorder are psychogenic and reflect a dominating effect of disengagement coping strategies, which represent secondary compensatory adaptations during the chronic course of this disorder to counteract primary arousal symptoms, especially those related to an intractable shame-laden depressive syndrome. The psychoendocrine findings suggest that the relatively inconspicuous clinical feature of shame resulting from both the primary and secondary traumatizations is a particularly powerful, preoccupying, and overwhelming source of emotional engagement. Shame may represent a "sleeper" that is worthy of greater attention in both research and clinical efforts to understand the pathogenesis and psychopathology of this devastating stress related disorder. PMID- 11382268 TI - Relationship among plasma cortisol, catecholamines, neuropeptide Y, and human performance during exposure to uncontrollable stress. AB - OBJECTIVE: Although many people are exposed to trauma, only some individuals develop posttraumatic stress disorder; most do not. It is possible that humans differ in the degree to which stress induces neurobiological perturbations of their threat response systems, which may result in a differential capacity to cope with aversive experiences. This study explored the idea that differences in the neurobiological responses of individuals exposed to threat are significantly related to psychological and behavioral indices. METHODS: Individual differences in neurohormonal, psychological, and performance indices among 44 healthy subjects enrolled in US Army survival school were investigated. Subjects were examined before, during, and after exposure to uncontrollable stress. RESULTS: Stress-induced release of cortisol, neuropeptide Y, and norepinephrine were positively correlated; cortisol release during stress accounted for 42% of the variance in neuropeptide Y release during stress. Cortisol also accounted for 22% of the variance in psychological symptoms of dissociation and 31% of the variance in military performance during stress. CONCLUSIONS: Because dissociation, abnormalities in the hypothalamic-pituitary-adrenocortical axis, and catecholamine functioning have all been implicated in the development of stress disorders such as posttraumatic stress disorder, these data suggest that some biological differences may exist before index trauma exposure and before the development of stress-related illness. The data also imply a relationship among specific neurobiological factors and psychological dissociation. In addition, the data provide clues about the way in which individuals' psychobiological responses to threat differ from one another. PMID- 11382270 TI - Hostility, gender, and cardiac autonomic control. AB - OBJECTIVE: Although considerable evidence implicates hostility in the development of coronary artery disease (CAD), the pathogenic mechanisms remain poorly understood. We have developed a psychophysiological model that holds that altered autonomic nervous system function links psychological traits with CAD outcomes. In laboratory studies, stressors reduce high-frequency (HF) heart period variability, an index of cardiac vagal modulation. With ambulatory electrocardiographic recording, we demonstrated in a predominantly male sample that hostility was inversely associated with HF power, but only during waking hours. These findings are consistent with the hypothesis that hostile individuals experience multiple stressful interpersonal transactions each day, resulting in overall lower HF power during the day but not at night. METHODS: To further evaluate this hypothesis, we screened 96 subjects using the Cook-Medley Hostility Scale and selected 15 men and 15 women representing a wide distribution of hostility. These subjects were studied in a laboratory session assessing reactivity to psychological and orthostatic challenges with continuous electrocardiographic, blood pressure, and respiration monitoring. We predicted that for men and women, hostility would be inversely related to reductions in HF power in response to challenge. RESULTS: In response to mental stressors, all measures of heart period variability change were inversely related to hostility as predicted. No such relationships were found for responses to tilt. The data suggested a possible effect of gender on these relationships. CONCLUSIONS: These data add to the growing body of evidence showing that hostility influences vagal modulation of the cardiovascular system and suggest that altered autonomic control is a pathogenic mechanism linking hostility and CAD. PMID- 11382269 TI - Partner interactions are associated with reduced blood pressure in the natural environment: ambulatory monitoring evidence from a healthy, multiethnic adult sample. AB - OBJECTIVE: The objective of this study was to examine the effects of partner interactions on ambulatory blood pressure in a sample of 120 healthy adults who were monitored over a 6-day period. METHODS: After each blood pressure measurement, participants rated characteristics of ongoing social interactions, along with emotional activation, physical activity, talking, posture, and other covariates, with computer-assisted self-report assessments. RESULTS: Using multilevel modeling, we showed that blood pressure was significantly lower during social interactions with one's partner relative to social interactions with any other person and relative to periods of not interacting. Interactions with partners also were characterized by significantly less talking and emotional activation and more intimacy and perceived emotional support; however, these differences did not mediate the partner effect on blood pressure. In addition, the relative benefits of interacting with a partner were not moderated by relationship quality, gender, or race. CONCLUSIONS: The effects of social situations on ambulatory blood pressure may represent one pathway through which social relationships affect cardiovascular health. Key words: blood pressure, social interaction, partner interactions, cardiovascular health. PMID- 11382271 TI - Somatic and psychological complaints and their correlates with insomnia in the Japanese general population. AB - OBJECTIVE: This cross-sectional study was conducted to estimate the prevalence of somatic and psychological complaints (SPCs) and to investigate the association of SPCs with insomnia in a sample of the general adult population of Japan. METHODS: We randomly selected 4000 adult residents (-20 years old) from five areas of Japan using stratified sampling and conducted interviews using a structured questionnaire. The questionnaire solicited information about eight somatic symptoms, eight psychological symptoms, three sleep problems, and demographic and health-related information. A total of 3,030 subjects completed questionnaires, giving a response rate of 75.8%. RESULTS: Stiff neck/shoulder (45.3%), backache (35.1%), and fatigue (31.4%) were the most common complaints in this population. In general, SPCs were more prevalent in younger persons and in women. Logistic regression analyses, controlling for other factors, showed that insomnia was significantly associated with a number of SPCs: backache (odds ratio [OR] = 1.4, 95% confidence interval [CI] = 1.1-1.6), epigastric discomfort (OR = 1.7, 95% CI = 1.3-2.2), weight loss (OR = 2.0, 95% CI = 1.2-3.3), headache (OR = 1.7, 95% CI = 1.3-2.2), fatigue (OR = 1.7, 95% CI = 1.4-2.1), worrying (OR = 1.6, 95% CI = 1.1-2.3), irritability (OR = 1.4, 95% CI = 1.1-1.7), and loss of interest (OR = 1.8, 95% CI = 1.2-2.7). CONCLUSIONS: SPCs were common and were largely associated with insomnia in the general adult population of Japan. Further study is needed to examine the causal links between SPCs and insomnia. PMID- 11382272 TI - Distraction reduces self-reported physiological reactions to blood donation in novice donors with a blunting coping style. AB - OBJECTIVE: Vasovagal reactions experienced by some blood donors (eg, faintness, lightheadedness, and dizziness) have been shown to be related to a decreased likelihood of future blood donations. This study evaluated the efficacy of audiovisual distraction as a means of reducing self-reported physiological reactions in first-time blood donors. Because interventions that are consistent with an individual's preferred coping style have been shown to be more effective at reducing physiological and psychological responses to stressful medical procedures, coping style (monitoring vs. blunting) was assessed as a possible moderating variable. METHODS: First-time blood donors were randomly assigned to one of two conditions: audiovisual distraction or no-treatment control. Participants in the distraction group donated blood at an American Red Cross blood drive while watching a three-dimensional video presentation on a personal visor and headset. The control group donated blood according to standard American Red Cross procedures. Score on a self-reported measure of physiological reactions completed immediately after donation served as the dependent variable. RESULTS: Individuals who typically use blunting coping strategies to cope with stress reported an attenuation of vasovagal reactions to blood donation in the distraction vs. the control condition (t(49) = 2.29, p < .05), whereas donors who prefer a monitoring coping style did not benefit from distraction. CONCLUSIONS: Among first-time blood donors, audiovisual distraction may be an effective means of reducing vasovagal reactions in donors who prefer to cope with stress using such strategies as distraction, denial, and reinterpretation. PMID- 11382273 TI - Pain and immunologic response to root canal treatment and subsequent health outcomes. AB - OBJECTIVE: This study examined the effects of pain and stress associated with a dental procedure, root canal treatment (RCT), on natural killer cell cytotoxicity (NKCC) and the subsequent development of symptoms of upper respiratory illness during the following month. METHODS: Patients (N = 33) were recruited from those scheduled for RCT appointments. Subjects for a non-RCT comparison group (N = 14) were also recruited from dental clinic patients. Peripheral blood was drawn by use of an indwelling catheter three times: just before RCT, 30 minutes after injection of a local anesthetic, and 30 minutes after RCT (a parallel time course was followed for the comparison group.) Blood was assayed for cortisol and NKCC. Subjects completed a health diary in the month after RCT. RESULTS: Patients showed a significant increase in NKCC between baseline and RCT and a significant decrease from RCT to after RCT, whereas the comparison group did not. The NKCC following the RCT was negatively correlated with the pain level during RCT (r = 0.48, p < .01) and pain levels 2 and 6 hours after RCT (r = -0.43, p < .05; r = 0.44 p < .05, respectively). The patient group reported significantly more illness episodes 2 weeks after RCT than the comparison group (Wilcoxon rank sum = 4.78, p = .03). Discriminant function analysis correctly classified 88% of the subjects into the illness category using predictor variables of post-RCT NKCC, stress, and pain levels during RCT (F(3,21) = 8.23, p < .001). CONCLUSIONS: Transitory changes in NKCC associated with pain and stress may be implicated in the development of infectious disease episodes after an acute stressful event. PMID- 11382274 TI - Defensive hostility and coronary heart disease: a preliminary investigation of male veterans. AB - OBJECTIVE: Research and theory link an interpersonal conflict model to cardiovascular disease. Specifically, persons scoring high on cynical hostility and social defensiveness are thought to manifest a defensive need for approval while harboring basic distrust and hostility toward those who could provide such approval. The objective of this study was to assess whether angiographically determined coronary artery disease (CAD) was associated with this combination of high cynical hostility and high social defensiveness. METHODS: Fifty-nine male patients of a Veterans Administration Medical Center (86% white, mean age = 59.9 years) participated in the study on the day before their angiographic procedure; these men filled out the Cook-Medley Hostility Scale and the Marlowe-Crowne Social Desirability Scale (social defensiveness). They subsequently were categorized as having defensive hostility (DH), high hostility (HH), high social defensiveness (SD), or low psychosocial risk (LRisk; low on both scales). RESULTS: The four groups did not differ significantly on risk factor status or health status. As predicted, a preplanned contrast showed that the DH group's mean number of arteries with at least 50% blockage (mean = 2.5) differed significantly from the combined means of the other groups. The HH and SD groups did not differ from the LRisk group. CONCLUSIONS: When combined with other reports, the approach-avoidance interpersonal conflict model holds the promise of providing additional information about the psychosocial factors contributing to CAD development among men with high cynical hostility. PMID- 11382275 TI - Anger inhibition potentiates the association of high end-tidal CO2 with blood pressure in women. AB - OBJECTIVE: High resting end-tidal CO2 (PetCO2) has been shown to be an independent predictor of systolic blood pressure (SBP) in women, particularly older women. The study reported in this article investigated whether the tendency to experience, express, and/or suppress anger contributes to the association of PetCO2 and SBP in women and in men. METHODS: The Spielberger Anger Expression Inventory was administered to 403 healthy male and female participants in the Baltimore Longitudinal Study on Aging. Resting PetCO2 was obtained by means of a respiratory gas monitor, and resting blood pressure was obtained with an oscillometric device. The associations of resting PetCO2 and the anger scales with SBP and diastolic blood pressure (DBP) were investigated using multivariate regression analyses. RESULTS: PetCO2, as well as age and body mass index, was an independent predictor of SBP in women with low, but not high, trait anger and in women with low, but not high, anger-out. PetCO2 was not an independent predictor of SBP in men with either high or low anger. In addition, PetCO2 was not an independent predictor of DBP in either men or women. CONCLUSIONS: The results of this study indicate that inhibition of anger potentiates the relationship of high PetCO2 with SBP in women but not men. Additional studies are needed to determine the origins of the observed gender differences and the psychophysiological pathways by which high resting PetCO2 contributes to elevated resting blood pressure in women. PMID- 11382277 TI - Normalization of hypertensive responses during ambulatory surgical stress by perioperative music. AB - OBJECTIVE: The purpose of this study was to determine whether cognitive appraisals of stress level and hypertensive responses to ambulatory ophthalmic surgery can be ameliorated by patient-selected music. METHODS: We studied 40 elderly individuals requiring ophthalmic surgery, 20 in an experimental group (mean age, 74 years) and 20 in a control group (mean age, 77 years). All patients had an established resting blood pressure <140/90 mm Hg. In the experimental group self-selected music was provided by headphones throughout the preoperative, surgical, and postoperative periods. In the control group patients had neither headphones nor music. All patients received similar (weight-determined) doses of alfentanil and midazolam during surgery. Heart rate, blood pressure, and patient reported stress and coping levels were the dependent variables. RESULTS: In both groups, blood pressure values were normal (approximately 129/82 mm Hg) during screening examinations 1 week before surgery. On the day of surgery both groups displayed increased preoperative blood pressures (approximately 159/92 mm Hg) associated with increases in heart rate (by approximately 17 beats/min). Intraoperative blood pressures in the experimental group returned quickly to screening baseline values, whereas the control group experienced persistent elevations in intraoperative blood pressure similar to preoperative levels. Over the course of the surgical experience, patients with music reported significant reductions in perceived stress and increases in coping abilities (p < .001), whereas those without music did not. CONCLUSIONS: The perceived stress of ambulatory surgery in geriatric patients is associated with a clinical hypertensive response that is ameliorated by self-selected perioperative music, which also decreases perceived stress and increases patients' sense of personal control and well-being. PMID- 11382276 TI - Mechanisms and mediators of psychological stress-induced rise in core temperature. AB - OBJECTIVE: Despite numerous case reports on "psychogenic fever," it remains uncertain how psychological stress raises core temperature and whether the rise in core temperature is a real fever or a hyperthermia. This article reviews studies on the psychological stress-induced rise in core temperature (PSRCT) in animals with the aim to facilitate studies on the mechanisms of so-called psychogenic fever in humans. METHODS: To address this question, we reviewed the mechanisms and mediators of the PSRCT and classic conditioning of the fever response in animals. RESULTS: The PSRCT is not due to the increased locomotor activity during stress, and the magnitude of the PSRCT is the same in warm and cold environments, indicating that it is a centrally regulated rise in temperature due to an elevated thermoregulatory "set point." The PSRCT caused by conventional psychological stress models, such as open-field stress, is attenuated by cyclooxygenase inhibitors, which block prostaglandin synthesis. On the other hand, the PSRCT elicited by an "anticipatory anxiety stress" is not inhibited by cyclooxygenase inhibitors but by benzodiazepines and serotonin Type 1A receptor agonists. The febrile response can be conditioned to neutral stimuli after paired presentation with unconditioned stimuli such as injection of lipopolysaccharide, a typical pyrogen. CONCLUSIONS: Most findings indicate that the PSRCT is a fever, a rise in the thermoregulatory set point. The PSRCT may occur through prostaglandin E2-dependent mechanisms and prostaglandin E2 independent, 5-HT-mediated mechanisms. The febrile response can be conditioned. Thus, these mechanisms might be involved in psychogenic fever in humans. PMID- 11382278 TI - Heart rate, neuroendocrine, and immunological reactivity in response to an acute laboratory stressor. AB - OBJECTIVE: The primary objective of the present study was to identify neuroendocrine and immunological correlates of cardiovascular reactivity to an acute laboratory stressor. METHODS: Subjects were 56 healthy volunteers. Heart rate and blood pressure were assessed at regular intervals during a 30-minute adaptation period and a 6-minute videotaped speech task. Blood was drawn before and after the task and was assayed for natural killer cell activity (NKCA), cortisol production, in vitro interferon gamma (IFN-gamma) and interleukin 10 production by peripheral blood mononuclear cells (PBMC), and antibody titers to the Epstein-Barr virus. Psychological measures were also administered. RESULTS: NKCA increased significantly in response to the task, and this increase was significantly and positively correlated with heart rate reactivity. IFN-gamma production by PBMC also increased in response to the task, but these increases were unrelated to heart rate reactivity. In addition, baseline cortisol levels were found to be predictive of heart rate reactivity. Finally, questionnaire data were modestly related to various aspects of stress-induced reactivity. CONCLUSIONS: Consistent with the task-related increases in NKCA and IFN-gamma, acute stress may signal an increase in at least some aspects of the cell mediated, or TH1-driven, immune response. Furthermore, the finding that heart rate reactivity was related in part to baseline individual differences in cortisol production suggests that short-term cardiovascular responses to stress may be directly related to longer-term neuroendocrine modulation. Finally, the present results also help to highlight the influence of both sympathetic and nonsympathetic pathways in the response to acute stressors and suggest tentative links between certain psychological traits and various aspects of stress-induced reactivity. PMID- 11382279 TI - Cytokines and anorexia nervosa. AB - OBJECTIVE: Recent studies have indicated that the inflammatory cytokines could be implicated in anorexia nervosa and in its complications. To determinate the potential role of interleukins (IL-1, IL-2, IL-4, IL-6, IL-10), interferon (IFN gamma), tumor necrosis factor (TNF-alpha), and transforming growth factor (TGF beta2) in anorexia nervosa, serum concentrations of these cytokines were measured in patients suffering from anorexia nervosa in comparison to healthy subjects. METHOD: Twenty-nine anorexic women according to DSM-IV criteria participated in the study. The control group consisted of 20 healthy women without eating disorders, mood disorders, and immunological disorders. RESULTS: We find that serum IL-2 and TGF-beta2 concentrations were both significantly decreased in anorexic patients, although the other cytokines did not differ significantly between the two groups. CONCLUSION: Our results show that in patients with anorexia nervosa, there are lower levels of specific cytokines (especially IL-2 and TGF-beta2). These levels may reflect the combination of impaired nutrition and weight loss, therefore, the dysregulation of these cytokines may contribute in anorexia's complications. Follow-up studies should examine the effects of parameters such as starvation, psychopathologic factors, and psychoneuroendocrinological perturbation which could affect interplay between cytokines, neuropeptides, and neurotransmitters. PMID- 11382280 TI - Sacral neuromodulation for urinary retention caused by sexual abuse. AB - OBJECTIVE: This case report describes the use of sacral neuromodulation to treat urinary retention after sexual abuse. METHODS: Sacral neuromodulation was added to therapeutic regimen of a 38-year-old woman in whom chronic, complete urinary retention developed after psychological and sexual abuse during childhood. RESULTS: The combination of psychotherapy and neuromodulation restored the patient's ability to void, whereas psychotherapy alone had not. CONCLUSIONS: Although a multifactorial etiology of retention cannot be ruled out in this patient, neuromodulation might effectively treat urinary retention in cases of a conversion disorder after sexual abuse. PMID- 11382281 TI - Influence of psychiatric diagnoses on the relationship between suicide attempts and the menstrual cycle. PMID- 11382282 TI - Conductivity detection and quantitation of isotachophoretic analytes on a planar chip with on-line coupled separation channels. AB - A poly(methylmethacrylate) chip, provided with two separation channels in the column-coupling (CC) arrangement and on-column conductivity detection sensors and intended, mainly, to isotachophoresis (ITP) and ITP-capillary zone electrophoresis (CZE) separations was developed recently. The present work was aimed at assessing its performance relevant to the detection and quantitation of the ITP analytes. Hydrodynamic (HDF) and electroosmotic (EOF) flows of the solution in the separation compartment of the CC chip were suppressed and electrophoresis was a dominant transport process in the ITP separations with model analytes carried out in this context. When the surfaces of the detection electrodes of the conductivity sensors on the chip were appropriately cleaned qualitative indices of the test analytes [relative step heights (RSHs)], provided by a particular detection sensor, agreed within 1% (expressed via RSDs of the RSH values). Their long-term reproducibilities for one sensor, as estimated from 70 ITP runs repeated in 5 days, were 2% or less. Sensor-to-sensor and chip-to-chip fluctuations of the RSH values for the test analytes were 2.5% or less. In addition, experimentally obtained RSH values agreed well with those predicted by the calculations based on the ITP steady-state model. Reproducibilities of the migration velocities attainable on the CC chips with suppressed EOF and HDF, assessed from the migration time measurements of the ITP boundary between well defined positions on the separation channels of the chips (140 repeated runs on three chips), ranged from 1.4 to 3.3% for the migration times in the range of 100 200 s. Within-day repeatabilities of the time-based zone lengths for the test analytes characterized 2% RSDs, while their day-to-day repeatabilities were less than 5%. Chip-to-chip reproducibilities of the zone lengths, assessed from the data obtained on three chips for 100 ITP runs, were 5-8%. PMID- 11382283 TI - Influence of buffer zone concentrations on efficiency in partial filling micellar electrokinetic chromatography. AB - The potential of counter pressure-moderated partial filling micellar electrokinetic chromatography (PF-MEKC) was investigated in this work. Plate numbers of homologous omega-phenylalcohols were measured in a two-plug PF-MEKC system varying the concentrations and hence the ionic strengths of the background buffer compared to the sodium dodecyl sulfate-containing separation buffer and the counter pressure on the cathodic buffer reservoir. It was observed that plate numbers are strongly influenced by both the buffer concentrations and the counter pressure. Highest plate numbers were obtained with a buffer system where the concentrations are adjusted such that the electroosmotic flow velocities in both zones are equal. Differences in the local electroosmotic flow velocities of the zones caused by different buffer concentrations are responsible for tremendously reduced plate numbers. The efficiency drop is explained in several models by the formation of an intersegmental pressure which produces a parabolically shaped laminar flow component in both zones. Thus, the electroosmotic plug-like flow profile is distorted and the efficiency is reduced. The effect of counter pressure on efficiency turned out to be very complex in dependence on the buffer system applied. PMID- 11382284 TI - On-line sample concentration in micellar electrokinetic chromatography using cationic surfactants. AB - Two on-line sample concentration techniques, sample stacking and sweeping, were evaluated using cationic surfactants as pseudostationary phases in micellar electrokinetic chromatography. As cationic surfactant micelles, tetradecyltrimethylammonium bromide and cetyltrimethylammonium chloride were employed. About 10-fold and 1000-fold increases in detection sensitivity in terms of peak heights were observed by sample stacking and sweeping, respectively, without suppression of the electroosmotic flow. In particular, the concentration limits of detection (S/N=3) for test naphthalenesulfonic acids obtained with sweeping were from 0.96 to 0.47 ppb with UV detection without any preconcentration procedure. PMID- 11382285 TI - New configuration in capillary isotachophoresis-capillary zone electrophoresis coupling. AB - The paper surveys possible configurations of a coupling capillary column operating in various electromigration modes. Special attention is given to capillary isotachophoresis-capillary zone electrophoresis (cITP-CZE) coupling and its description from the theoretical point of view. Computer simulations of separation are presented and compared with experiments. Further, we propose a new configuration of electrolyte systems in cITP-CZE coupling, which offers a possibility to perform complex analyses of micro- and macro-constituents in one run. The electrolyte system is verified by practical experiments for both anionic and cationic modes of analysis. The advantages and disadvantages of the new combination are discussed. PMID- 11382286 TI - Capillary zone electrophoresis of orotic acid in urine with on-line isotachophoresis sample pretreatment and diode array detection. AB - Potentialities of capillary zone electrophoresis with on-line isotachophoresis sample pretreatment and diode array detection (ITP-CZE-DAD) to the separation, detection and identification of trace analytes present in biological matrices were investigated. Urine represented a multicomponent, variable and high ionic strength matrix while orotic acid was chosen as a model analyte of a practical clinical relevance in this investigation. Using the ITP-CZE combination in the column-coupling configuration of the separation system ITP provided an enhanced sample load capacity to the separation system (a 30 microl sample injection volume), concentrated the analyte and served as an on-line sample clean up technique. On the other hand, CZE performed a final separation of the analyte from matrix constituents present in the ITP pretreated sample and provided favorable conditions for its detection and identification by DAD. Using current correction and smoothing procedures analytically relevant DAD spectra of orotic acid could be obtained also in instances when this was injected in a model sample at a 2 x 10(-7) mol/l concentration (an estimated limit of determination of orotic acid at a 218 nm detection wavelength). ITP-CZE separations of urine samples (based on differences in acid-base properties and host-guest complexations of the analyte and matrix anionic constituents) led to significant sample clean ups. Consequently, DAD spectra of orotic acid matching its reference spectrum, could be acquired also in instances when the acid was present in urine matrices (loaded in 30 microl injection volumes of 20-fold diluted urine samples) at 4-6 x 10(-7) mol/l concentrations. Here, residual trace matrix interferents prevented a closer approach to the above value attainable for model samples. Although this work was focused only on one analyte and urine matrix it implies very promising potentialities of the ITP-CZE-DAD combination in the identification and quantitation of trace analytes present in biological matrices, in general. PMID- 11382287 TI - Isotachophoresis and isotachophoresis--zone electrophoresis separations of inorganic anions present in water samples on a planar chip with column-coupling separation channels and conductivity detection. AB - The use of a poly(methylmethacrylate) chip, provided with two separation channels in the column-coupling (CC) arrangement and on-column conductivity detection sensors, to electrophoretic separations of a group of inorganic anions (chloride, nitrate, sulfate, nitrite, fluoride and phosphate) that need to be monitored in various environmental matrices was studied. The electrophoretic methods employed in this study included isotachophoresis (ITP) and capillary zone electrophoresis (CZE) with on-line coupled ITP sample pretreatment (ITP-CZE). Hydrodynamic and electroosmotic flows of the solution in the separation compartment of the CC chip were suppressed and electrophoresis was a dominant transport process in the separations performed by these methods. ITP separations on the chip provided rapid resolutions of sub-nmol amounts of the complete group of the studied anions and made possible rapid separations and reproducible quantitations of macroconstituents currently present in water samples (chloride, nitrate and sulfate). However, concentration limits of detection attainable under the employed ITP separating conditions (2-3 x 10(-5) mol/l) were not sufficient for the detection of typical anionic microconstituents in water samples (nitrite, fluoride and phosphate). On the other hand, these anions could be detected at 5-7 x 10(-7) mol/l concentrations by the conductivity detector in the CZE stage of the ITP-CZE combination on the CC chip. A sample clean-up performed in the ITP stage of the combination effectively complemented such a detection sensitivity and nitrite, fluoride and phosphate could be reproducibly quantified also in samples containing the macroconstituents at 10(4) higher concentrations. ITP-CZE analyses of tap, mineral and river water samples showed that the CC chip offers means for rapid and reproducible procedures to the determination of these anions in water (4-6 min analysis times under our working conditions). Here, the ITP sample pretreatment concentrated the analytes and removed nanomol amounts of the macroconstituents from the separation compartment of the chip within 3-4 min. Both the ITP and ITP-CZE procedures required no or only minimum manipulations with water samples before their analyses on the chip. For example, tap water samples were analyzed directly while a short degassing of mineral water (to prevent bubble formation during the separation) and filtration of river water samples (to remove particulates and colloids) were the only operations needed in this respect. PMID- 11382288 TI - Determination of organic acids and inorganic anions in wine by isotachophoresis on a planar chip. AB - Isotachophoretic (ITP) separation and determination of a group of 13 organic and inorganic acids, currently present in wines, on a poly(methyl methacrylate) chip provided with on-column conductivity detection was a subject of a detailed study performed in this work. Experiments with the ITP electrolyte systems proposed to the separation of anionic constituents present in wine revealed that their separation at a low pH (2.9) provides the best results in terms of the resolution. Using a 94 mm long separation channel of the chip, the acids could be resolved within 10-15 min also in instances when their concentrations corresponded to those at which they typically occur in wines. A procedure suitable to the ITP determination of organic acids responsible for some important organoleptic characteristics of wines (tartaric, lactic, malic and citric acids) was developed. Concentrations of 2-10 mg/l of these acids represented their limits of quantitation for a 0.9 microl volume sample loop on the chip. A maximum sample load on the chip, under the preferred separating conditions, was set by the resolution of malate and citrate. A complete resolution of these constituents in wine samples was reached when their molar concentration ratio was 20:1 or less. ITP analyses of a large series of model and wine samples on the chip showed that qualitative indices [RSH (relative step height) values] of the acids, based on the response of the conductivity detector, reproduced with RSD better than 2% while reproducibilities of the determination of the acids of our interest characterized RSD values better than 3.5%. PMID- 11382289 TI - Determination of 5-aminosalicylic acid related impurities by micellar electrokinetic chromatography with an ion-pair reagent. AB - A micellar electrokinetic chromatographic (MEKC) method was developed for the quantification of mesalazine or 5-aminosalicylic acid (5-ASA) and its major impurities 3-aminosalicylic acid, salicylic acid, sulfanilic acid and 4 aminophenol. The optimisation of the experimental conditions was carried out considering some important requirements: resolution, reproducibility, detection limits of 0.1% (m/m) or less, short total analysis time. Preliminary investigations employing sodium dodecyl sulfate (SDS) as surfactant did not lead to the necessary resolution of the studied compounds; the addition of tetrabutylammonium bromide (TBAB) to the SDS micellar system resulted in the complete separation of all the compounds. The effects on the separation by several parameters such as TBAB concentration, SDS concentration, background electrolyte pH and concentration, were evaluated. Using a fused-silica capillary (8.5 cm effective length) complete analysis was obtained in a very short time. Under the optimised final conditions [120 mM 3-(cyclohexylamino)-2-hydroxy-1 propanesulfonic acid buffer, pH 10.20, 65 mM SDS in the presence of 55 mM TBAB and 5% methanol] the method was validated for specificity, precision, linearity, limits of detection and quantitation, and robustness: the 5-ASA related impurities can be quantified at least at the 0.1% (m/m) level. PMID- 11382290 TI - Application of capillary isotachophoresis for fruit juice authentication. AB - A method using capillary isotachophoresis (ITP) was developed and applied for the determination of the anionic profile of orange juices with the aim to obtain some useful information on the authenticity or adulteration of imported and native beverage products. An EA 100 electrophoretic analyser (Villa-LABECO, Slovak Republic) was used for capillary isotachophoretic determination of anions in tested samples. More systems of leading and terminating electrolytes were used. Detection conductivity and UV detection at 254 nm were used. Sample injection volume was 30 microl. These systems allow one to determinate inorganic anions, organic acids and some additives--adulterants in anionic forms in orange juices. By capillary isotachophoretic determination the lengths or areas of characteristic zones were established and compared to authentic orange juices of different species and origin and with RSK reference values (Code of Practice). Special emphasis was placed on D-isocitric acid ITP determination as a reliable fruit juice authentication marker. The presented multicomponent analysis of orange juice authenticity according to ITP anionic profiles obtained by capillary isotachophoresis presents an alternative information source necessary for deciding about authenticity of the products. PMID- 11382291 TI - Discrete spacers for photometric characterization of humic acids separated by capillary isotachophoresis. AB - A group of twenty discrete spacers suitable for photometric characterization of humic acids (HAs) after their isotachophoretic (ITP) separation at pH 10 was found. The spacers, inorganic and organic acids and amino acids of suitable acid base and migration properties exhibiting no light absorption in the UV region of the light spectrum, made possible to perform this characterization in a sensitive spike mode of the ITP analysis. Using this approach a complex mixture of humic constituents present in a test HA preparation was separated into 22 fractions migrating in the interzonal boundary layers formed by the zones of discrete spacers and 21 fractions mixed with the zones of the spacers. A photometric monitoring of the fractions in the ITP stack at a 405 nm detection wavelength provided an adequate selectivity and sensitivity into the characterization. Relative sizes of the detected fractions of the test HA preparation ranged from 0.2-0.3 to 27.5% (based on the response of the phototometric detector at 405 nm). The fractions representing ca. 0.2-0.3% of the total peak area could be still quantified when 800 ng of the test preparation was loaded onto the ITP column. A typical repeatability of the total area of the detection signal corresponding to humic constituents in the ITP stack was ca. 2.5%. Repeatabilities of the peak areas of the fractions of the humic constituents defined by the spacers ranged from 2 to 6% for the fractions representing 1% or more of the total area and from 8 to 12% for those representing less than 1%. No marks of aggregations of the humic constituents were detected and reproducible ITP profiles (fingerprints) of the studied humic preparation were achieved under the developed working conditions. PMID- 11382292 TI - Analysis of proguanil and its metabolites by application of the sweeping technique in micellar electrokinetic chromatography. AB - The method of applying large sample volumes in micellar electrokinetic chromatography termed sweeping is applied to determine the conservative limits of detection of some basic drugs in plasma and urine. The biguanides proguanil, 4 chlorophenylbiguanide and cycloguanil are used as models of basic drugs and the limits of detection obtained compared with those previously reported for capillary zone electrophoresis using field-amplified sample injection (FASI) and also by LC using off-line preconcentration. It is found that the sweeping method can be applied to extracts of such biological matrices. The limits of detection obtained by sweeping are improved over FASI for plasma but not for urine and the limits of detection are higher than those reported for LC, for these compounds. PMID- 11382293 TI - Isotachophoretic determination of naproxen in the presence of its metabolite in human serum. AB - An isotachophoretic method with conductivity detection was developed to determine naproxen in the presence of its metabolite 6-O-desmethylnaproxen in human serum. The leading electrolyte contained 10 mM hydrochloric acid, beta-alanine, pH 4.0 and 0.1% methylhydroxypropylcellulose. The terminating electrolyte was 10 mM 2-(N morpholino)ethanesulfonic acid-tris(hydroxymethyl)aminomethane, pH 6.9, containing 20% (v/v) of ethanol. Naproxen was determined in serum supernatant after simple deproteination of the sample with ethanol. The isotachophoretic results were compared with those obtained by synchronous fluorescence spectrometry. PMID- 11382294 TI - Determination of 5-fluorouracil and 5-fluoro-2'-deoxyuridine-5'-monophosphate in pancreatic cancer cell line and other biological materials using capillary electrophoresis. AB - Capillary zone electrophoresis (CZE) was used for the rapid determination of 5 fluorouracil (5-FU) and 5-fluoro-2'-deoxyuridine-5'-monophosphate (FdUMP) in pancreatic cancer cell line (PANC-1), culture medium, plasma and pancreatic tissue. The assay is based upon protein precipitation with acetonitrile followed by a 9-min CZE analysis of the supernatant in an uncoated fused-silica capillary employing a borate buffer and on-column absorbance detection at 265 nm. Using 50 microl of sample, 5-FU levels between 4.12 and 132 microg/ml (31.7-1000 microM) were found to provide linear calibration graphs. Intra-day and inter-day RSD values evaluated from peak height ratios (n=5) were <7.6 and <8.8%, respectively. Corresponding RSD values of detection times (n=7) were <1 and <1.5%, respectively. The limits of detection for 5-FU and FdUMP were 1.72 and 5.29 microg/ml, respectively. As application, the accumulation of 5-FU by PANC-1 cells over a 4-h time period was investigated. Having a culture medium concentration of 100 microg/ml, the 5-FU cell content was estimated to become equal to that of the surrounding medium (i.e., 100 microg/ml or 3.61 fmol per cell with a volume of 4.7 pl) within that time period. The sensitivity of the assay was sufficient for the determination of 5-FU in all cell samples. FdUMP, however, could not be detected in these samples. Furthermore, the data obtained in uncoated capillaries are compared to those measured in a fused-silica capillary whose inner surface was coated with linear polyacrylamide (about 10-fold reduction of electroosmosis). The latter capillary format was found to be useless for simultaneous analysis of 5-FU and FdUMP in pancreatic cells but could be potentially useful for analysis of these compounds in plasma. PMID- 11382295 TI - Capillary electrophoresis-electrospray ionization ion trap mass spectrometry for analysis and confirmation testing of morphine and related compounds in urine. AB - Using an aqueous background electrolyte containing 25 mM ammonium acetate and NH3 (pH 9), CE-tandem MS and CE-triple MS with atmospheric pressure electrospray ionization in the positive ion mode are shown to represent attractive approaches for analysis and confirmation testing of morphine (MOR) and related opioids in human urine. Injection of plain or diluted urine permits monitoring of solutes at concentrations above 2-5 microg/ml. For the recognition of lower concentrations, solute extraction and concentration is required. Liquid-liquid extraction at alkaline pH is shown to be suitable for analysis of free opioids only whereas solid-phase extraction using a mixed-mode polymer phase is demonstrated to permit analysis of both free and glucuronidated opioids. The former sample preparation approach, however, requires about half of the time only. Commencing with 2 ml of urine, reconstitution to provide a sample volume of 0.2 ml and hydrodynamic sample injection, detection limits for free opioids are shown to be on the 100 200 ng/ml drug level. Much improved (ppb) sensitivity is obtained by infusing the extract directly into the source of the MS system. However, solutes that produce equal fragments (such as the two glucuronides of MOR) can thereby not be distinguished. CE-tandem MS and CE-triple MS are demonstrated to be suitable to confirm the presence of MOR, MOR-3-glucuronide, 6-monoacetylmorphine, codeine, codeine-6-glucuronide, dihydrocodeine, methadone and 2-ethylidene-1,5-dimethyl 3,3-diphenylpyrrolidine in a toxicological quality control urine. The same is shown for selected metabolites of codeine and dihydrocodeine in urines collected after administration of pharmaceutical preparations. PMID- 11382296 TI - On-line concentration of s-triazine herbicides in micellar electrokinetic chromatography using a cationic surfactant. AB - On-line concentration of neutral species of s-triazine herbicides in micellar electrokinetic chromatography using tetradecylammonium bromide (TTAB) as a cationic surfactant was investigated. Factors affecting the stacking of analytes were examined. The results indicate that the stacking efficiency is markedly improved with addition of phosphate buffer in the sample matrix. It was found that, depending on the nature of the analytes, the most effective stacking of these analytes occurs when the ratio of the conductivity of buffer electrolyte to that of sample matrix is in the range 1.4-1.2, with sample matrix containing phosphate buffer. Micelle concentration in the separation buffer is also a crucial factor to enhance the stacking efficiency and detection sensitivity of analytes. Moreover, the stacking efficiency of each individual analyte depends on its binding constant to TTAB micelles. The concentration effect is primarily based on sweeping mechanism which is operated in a normal stacking mode with reversed electrode polarity in the presence of reversed electroosmotic flow. As a result of concentration enhancement, the detection limits of these herbicides can reach about 9-15 ng/ml with UV detection. PMID- 11382297 TI - Analysis of metabolites of glucose pathways in human erythrocytes by analytical isotachophoresis. AB - The aim of this study was to establish an isotachophoretic (ITP) method for the determination of the main compounds of glycolysis in human erythrocytes in order to analyze the influence of different glucose concentrations (mimicking the situation in diabetes mellitus) on this pathway. Samples for ITP were prepared by isolation of erythrocytes, lysis of the cells by heating in double-distilled water and subsequent ultrafiltration (Mr cut-off: 5000). All the main compounds of glycolysis were characterized by ITP. The influence of different glucose concentrations on the main compounds of the energy metabolism (ATP, ADP, lactate, pyruvate) and 2,3-diphosphoglycerate were analyzed in short- and long-time incubations. PMID- 11382298 TI - Software and internet resources for capillary electrophoresis and micellar electrokinetic capillary chromatography. AB - The purpose of the contribution is to show how software and the internet have changed the way in which we carry out research, and what additional possibilities these new resources and tools provide. The internet and e-mail broaden our horizon for cooperation. There are no borders for information exchange: our library is a virtual one, electronic databases are at our fingertips. E-mail discussion groups provide an electronic community of CE users. Such forums have provided a basis for worldwide scientific cooperation amongst scientists; the present contribution is only one of several examples. Estimation software provides us with estimates of component properties in those cases where this information is not available from literature or from experiments (an estimated value is better than no value). Several examples will illustrate the use of estimation software in capillary zone electrophoresis and micellar electrokinetic capillary chromatography. Simulation software presents visualization of experimental results to be expected, both as a training tool and as the first step in method development. Other simulations yield valuable insights into phenomena that are not readily accessible experimentally for reasons of size or time-scale. PMID- 11382299 TI - Chemometric modeling of neurotransmitter amino acid separation in normal and reversed migration micellar electrokinetic chromatography. AB - A chemometric experimental design has been applied for the optimization of neurotransmitter amino acid separation in capillary electrophoresis. The optimizations were carried out for normal micellar electrokinetic chromatography (N-MEKC) and reversed migration micellar electrokinetic chromatography (RM-MEKC). In order to optimize three separation factors and study the interaction between factors, a response function was optimized via searching its optimum (minimum/maximum). For this purpose a central composite design with multivariate linear regression (MLR) analysis was utilized. Modeling with good regression coefficients from the MLR adequately described the interaction of factors such as background electrolyte and sodium dodecylsulfate concentrations which had a large impact on selectivity and migration behaviors. Similar optimal conditions regarding resolution and number of theoretical plates but different retention behaviors as a function of background electrolyte and micellar concentrations were observed for N-MEKC and RM-MEKC. Improved overall performance from the RM MEKC separation of five neurotransmitter acids, superior to N-MEKC, is demonstrated in terms of repeatability, peak symmetry, sensitivity, and in particular, impurity determination in an overloaded separation system. PMID- 11382300 TI - Application of capillary electrochromatography using macroporous polyacrylamide columns for the analysis of lignans from seeds of Schisandra chinensis. AB - Capillary electrochromatography (CEC) using polymer-based monolithic stationary phase has been developed as a promising method for the determination of lignans of Schisandra chinensis. The columns were prepared by in situ copolymerisation of acrylamide, N,N'-methylenebisacrylamide, vinylsulfonic acid and lauryl acrylate in presence of poly(ethylene glycol) as a porogenic agent. The columns [33 cm (24.5 cm effective length) x 75 microm I.D.] were successfully used to analyse and quantify the major lignans in extract of the seeds of Schisandra chinensis. Good separations were achieved in less than 35 min. The calibration graphs were linear in the range 0.025-1.0 mg/ml of given lignan with correlation coefficients between 0.9951 and 0.9996. The inter-day reproducibility of the peak area were below 3.9% and the inter-day reproducibility of the migration time were below 4.2%. The results of quantitative CEC analyses were compared with those obtained by reversed-phase HPLC, the levels of schizandrin, gomisin A, gomisin N and wuweizisu C determined by CEC were in a good agreement with those determined by HPLC. PMID- 11382301 TI - Analysis of thiocyanate in biological fluids by capillary zone electrophoresis. AB - A new sensitive and simple method has been developed for the determination of thiocyanate in human serum, urine and saliva. The determinations were performed in a fused-silica capillary [64.5 cm (56 cm effective length) x 75 microm] using 0.1 M beta-alanine-HCl (pH 3.50) as a background electrolyte, separation voltage 18 kV (negative polarity), temperature of capillary 25 degrees C and direct detection at 200 nm. Serum samples were 10-times diluted with deionised water and deproteinised with acetonitrile in the ratio 1:2. Urine and saliva samples need only 20-fold dilution with deionised water. The proposed method was successfully applied to the determination of thiocyanate in various human serum, saliva and urine samples. PMID- 11382302 TI - Capillary zone electrophoresis applied to the determination of the angiotensin converting enzyme inhibitor cilazapril and its active metabolite in pharmaceuticals and urine. AB - A capillary zone electrophoresis method has been developed for the quantitation of antihypertensive drug cilazapril and its active metabolite cilazaprilat in pharmaceuticals and urine. The separation of the compounds was performed in a fused-silica capillary filled with the running electrolyte, which consisted of a 60 mM borate buffer solution at pH 9.5. Under the optimized experimental conditions, the separation took less than 5 min. The analysis of urine samples required a previous solid-phase extraction step using C8 cartridges. The method was successfully applied to the determination of the drug and its metabolite in urine samples obtained from three hypertensive patients (detection limits of 115 ng ml(-1) for cilazaprilat and 125 ng ml(-1) for cilazapril) and to pharmaceutical dosage forms. The method was validated in terms of reproducibility, linearity and accuracy. PMID- 11382303 TI - Rapid capillary electrophoretic method for the determination of clozapine and desmethylclozapine in human plasma. AB - A rapid and sensitive high-performance capillary electrophoretic method for the determination of clozapine and its main metabolite desmethylclozapine in human plasma was developed. The separation of the two analytes was carried out in an untreated fused-silica capillary [33 cm (8.5 cm effective length) x 50 microm I.D.] filled with a background electrolyte at pH 2.5 containing beta cyclodextrin. Baseline separation of clozapine and desmethylclozapine was recorded in less than 3 min. An accurate sample pretreatment by means of solid phase extraction and subsequent concentration allows for reliable quantitation of clozapine in the plasma of schizophrenic patients under treatment with the drug. The method showed good precision (mean RSD = 4.0%) as well as satisfactory extraction yields (approximately 88%) and a good sensitivity (limit of quantitation = 0.075 microg ml(-1), limit of detection = 0.025 microg ml(-1)). PMID- 11382304 TI - Determination of the beta-blocker atenolol in plasma by capillary zone electrophoresis. AB - A capillary zone electrophoretic method was optimised for the determination of the beta-blocker atenolol in plasma. Separation was performed in an uncoated silica capillary of 58.5 cm (effective length 50 cm) x 75 microm I.D., and detection was at 194 nm. The effects of the buffer (concentration and pH), the injection time, the voltage applied and the plasma clean-up procedure were studied. The determination of atenolol was achieved in less than 3 min, using an electrolyte of 50 mM H3BO3-50 mM Na2B4O7 (50:50, v/v) pH 9, injected hydrodynamically for 4 s at 50 mbar and applying a voltage of +25 kV. This method was applied to the determination of atenolol in plasma of nine hypertensive patients (male and female, aged from 39 to 73 years). Atenolol concentrations found vary from 30 to 585 ng/ml. PMID- 11382305 TI - Theory of capillary electrochromatography. AB - The present state of the theory of capillary electrochromatography (CEC) is reviewed. Emphasis is placed on electroosmosis and the electrical double layer, and the generally good understanding of the factors affecting the electroosmotic flow in CEC columns. The relation of CEC to other electrically driven separations are described, along with band broadening, and the influence of column temperature in CEC. The theoretical potential of CEC is assessed from the standpoint of current and future column technology, and likely future application areas are described. PMID- 11382306 TI - Effect of temperature on the separation of long DNA fragments in polymer solution. AB - Electrophoresis of long DNA fragments in polymer solutions is still attractive when performed in short capillaries. Then the separations can be accomplished in minutes rather than hours as is usual in various slab electrophoresis techniques. In this paper we focused on the behavior of large DNA fragments in pulsed field capillary electrophoresis under various temperature conditions. The mobility dependence of fragments of lambda-DNA single-cut mixture on various frequencies at three different temperatures showed that the antiresonance mobility minima are shifted to higher frequencies at higher temperatures. This interesting result is explained in terms of the geometration model of DNA motion. PMID- 11382307 TI - Conductivity detection cell for capillary zone electrophoresis with a solution mediated contact of the separated constituents with the detection electrodes. AB - A contact conductivity detection cell for capillary zone electrophoresis (CZE) with an electrolyte solution mediated contact of the separated constituents with the detection electrodes (ESMC cell) was developed in this work. This new approach to the conductivity sensing in CZE is intended to eliminate detection disturbances due to electrode reactions and adsorption of the separated constituents when these are coming into direct contact with the detection electrodes. An optimum detection performance of the cell was achieved when the carrier electrolyte solution mediated the electric contact of the detection electrodes with the separated constituents. Different compositions of the mediator and carrier electrolyte solutions led to large drifts of the detection signals. Isotachophoresis experiments performed in this context with the ESMC cell revealed that origins of these drifts are in transport processes (diffusion and electromigration) between the detection compartment and the detection electrodes in the cell. These processes affected, to some extent, other analytically relevant performance parameters of the ESMC cell of the present construction as well [e.g., concentration limits of detection (LODs), a contribution of the cell to the band broadening]. For example, the ESMC cell gave, under optimum operating conditions, 3-4 times higher concentration LODs for the test analytes than a current on-column conductivity cell employed under identical working conditions. On the other hand, these LOD values (25-150 nmol/l) were still 20-25 times lower than those estimated from reference experiments for a contactless conductivity detector. CZE experiments with iodide, carried out under working conditions leading to electrochemical reactions of this anion on the detection electrodes of current conductivity cells, did not occur in the ESMC cell. In addition, this cell, contrary to a reference contact conductivity cell, required no special care (e.g., cleaning of the surfaces of the detection electrodes by chemical or electrochemical means) to maintain its reliable long term performance. Anionic CZE analyses of tap and mineral water samples monitored by the conductivity detector provided with the ESMC cell demonstrated a practical applicability and certain limitations of this detection approach in the analysis of ionic constituents present in high ionic strength sample matrices. PMID- 11382308 TI - Why robust background electrolytes containing multivalent ionic species can fail in capillary zone electrophoresis. AB - In this paper it is demonstrated that system peaks are induced by multivalent weak ionic species in background electrolytes. Their existence is derived from SystCharts and from Peak Shape Diagrams and the theory is confirmed experimentally. If analytes are present in a sample, with mobilities approximately equal to the mobility of a system peak, they interact, resulting in a strong increase of electromigration dispersion. This leads to strong peak broadening, peak deformation and a loss of resolution. Typical background electrolytes containing multivalent ionic species, e.g. phosphate and phthalate buffers, often reported to be robust electrolytes, are therefore not always universally applicable and can fail for the application of specific analytes. This paper reports a systematic study of the above phenomena and shows both theoretical and experimental results for background electrolytes containing phosphoric acid and phthalic acid. PMID- 11382309 TI - Lonene-coated sulfonated silica as a packing material in the packed-capillary mode of electrochromatography. AB - An aliphatic ionene (2-10-ionene) has been selected as a modifier to prepare a novel polymer-coated packing material for capillary electrochromatography. The packing material was produced by dynamical modification of a commercially available sulfonated silica Exsil-100 SCX. Strong ion-exchange interactions in the capillary packed with ionene-modified sulfonated silica have been demonstrated by the example of the retention of p-aminobenzoic acid. The total calculated anion-exchange capacity of the sorbent in the capillary was about 4 x 10(-9) mol. A fast separation (about 15 min) of several aromatic acids was achieved with the packing material. The highest number of theoretical plates obtained was about 120,000. Limits of detection of the aromatic acids were 2-5 microg/ml. The advantages and lacks of the approach are discussed briefly. PMID- 11382310 TI - Hemispherodextrins, a new class of cyclodextrin derivatives, in capillary electrophoresis. AB - A capped cyclodextrin derivative (THCMH), called hemispherodextrin, was observed to behave as a very efficient chiral selector for a variety of phenoxyacid enantiomeric pairs, both at pH 6 and pH 9. The very low concentration necessary to obtain separation was particularly impressive. The behaviour of THCMH was compared with that of other hemispherodextrins and cyclodextrin derivatives and the conclusions are reported. Some interesting conclusions are drawn by comparing the behaviour of THCMH with that of other hemispherodextrins reported elsewhere. PMID- 11382311 TI - Capillary isoelectric focusing with UV-induced fluorescence detection. AB - Low-molecular-mass fluorescent compounds excitable in the near UV region with suitable acidobasic and electrophoretic properties are suggested as isoelectric point (pI) markers for isoelectric focusing (IEF) with UV photometric and UV excited fluorometric detection. The experimental set-up of capillary IEF with UV excited fluorometric detection and properties of new UV-induced fluorescent pI markers are given. The pI values of 18 new pI markers determined independently of IEF methods range from 2.1 to 10.3. The examples of separation of new pI markers together with derivatized proteins by capillary IEF with photometric or fluorometric detection are presented. PMID- 11382312 TI - Use of a fluorosurfactant in micellar electrokinetic capillary chromatography. AB - A fluorosurfactant, the anionic N-ethyl-N-[(heptadecafluorooctyl)sulfonyl]glycine potassium salt, trade name FC-129 [CAS 2991-51-7] was investigated for possible application in micellar electrokinetic capillary chromatography (MEKC). The surfactant was characterized with conductometric titration and test sample mixtures were investigated in MEKC systems, and compared with sodium dodecylsulphate. An increased efficiency and interesting selectivity differences were observed. PMID- 11382313 TI - Electrokinetic chromatography with micelles, polymeric and monomeric additives with similar chemical functionality as pseudo-stationary phases. AB - A comparison is made of the retention properties of additives applied as positively charged pseudo-stationary phases for electrokinetic chromatography of neutral analytes. All additives have a quaternary ammonium as functional group. The polymeric additive [poly(N,N,N',N'-tetramethyl-N trimethylenehexamethylenediammonium), Polybrene] has a concentration of 2% (w/w) in the background electrolyte (acetate, pH 5.2). Monomeric octyltrimethylammonium (OTMA) was used at a concentration below or above its critical micelle concentration (CMC) (140 mmol/l). At a concentration (259 mmol/l) above the CMC the system is that normally used for micellar electrokinetic chromatography with cationic micelles. However, even below the CMC, where OTMA is present as monomer, retention of the neutral analytes is observed as well. In all systems coating of the capillary wall with Polybrene establishes an electroosmotic flow directed towards the anode, counter-migrating to the electrophoretic movement of the additive. Based on the measurement of the mobility of the analytes (15 small, monofunctional aromatic compounds with different functional groups), their capacity factors, k(i), were determined in all systems. Low correlation of the k(i) values is observed between the particular systems, indicating their different selectivity at least for individual pairs of analytes. Based on the log k(i) values, a linear free energy relationship was applied to elucidate the main types of chemical interaction responsible for retention. As a result, cavity formation and n or pi electron interactions were found being significant for the micellar OTMA system, which agrees with findings described in the literature for other (cationic and anionic) micellar systems. For the polymeric system and for the monomeric OTMA system, the significant retention parameter is indicating n and pi electron interactions. PMID- 11382314 TI - Extremely high electric field strengths in non-aqueous capillary electrophoresis. AB - The influence of high electric field strength on the separation of basic analytes in non-aqueous alcohol background electrolyte (BGE) solutions was investigated. Increasing the separation voltage in capillary electrophoresis (CE) may be advantageous if the conductivity of the BGE solution is low enough to allow fast separations without excessive Joule heating or band broadening. The voltage range tested was 20-60 kV with methanol and ethanol, and 25-60 kV with propanol and butanol as solvent for BGE. The resulting electric field strengths ranged from 660 V cm(-1) to 2000 V cm(-1). Experiments were made with a special laboratory constructed CE instrument. The separation efficiency vs. voltage curve was found to vary with the alcohol BGE solution. The increase in voltage decreased the separation efficiency in the case of methanol BGE solution, but with the other BGEs a clear efficiency maximum was obtained above 30 kV. The highest separation efficiencies were achieved with propanol BGE solution, where the efficiency maximum was reached at 45 kV. However, reasonable efficiency was achieved even at 60 kV. The extent of Joule heating was determined by calculating the temperature inside the capillary and the observed plate heights were interpreted in terms of the Van Deemter equation. The decrease in the separation efficiency with higher voltage was attributed mainly to Joule heating in the case of methanol and ethanol BGE solution and to the analyte adsorption on the capillary wall with propanol and butanol BGE solutions. PMID- 11382315 TI - Surgical endoscopy--an old and new challenge for surgeons. PMID- 11382316 TI - Carcinogenesis of cancer of the papilla and ampulla: pathophysiological facts and molecular biological mechanisms. AB - BACKGROUND: Ampullary cancer has one of the highest resectability rates and best prognoses among neoplasms arising in the periampullary region. DISCUSSION: Early diagnosis due to early symptoms can partially explain the better prognosis as compared to other cancers of the periampullary region, but biologic factors should also be taken in account. In the past few years, the molecular mechanisms underlying this disease have been investigated and alterations of genes that regulate different cell functions have been described. Mutations of K-ras and of the tumor suppressor genes APC, p16 and p53 indicate a major disturbance in cell cycle regulation. CONCLUSIONS: If the molecular profile of ampullary cancer is examined in terms of rate and type of molecular changes, it seems to be more similar to intestinal than to pancreatic cancer. Furthermore, the fact that many ampullary carcinomas arise from adenomas and the frequent finding of ampullary tumors in patients affected by polyposis syndromes also suggest that ampullary and colon cancers share common molecular mechanisms of carcinogenesis. PMID- 11382317 TI - Adenoma of the papilla and ampulla--premalignant lesions? AB - Ampullary adenomas arising in the papilla or the ampulla Vateri, are rare, benign, neoplastic lesions. No specific aetiological factors, such as diet, chemical or environmental causes, have been identified yet. An established risk factor which is accompanied by the development of adenoma is the presence of genetically inherited polyposis syndromes, e.g. familial adenomatosis coli (FAP). Adenomas assume tubular, tubulovillous, or villous architecture and are not different from adenomas arising elsewhere in the gastrointestinal tract. The full neoplastic spectrum, ranging from mild to high grade dysplasia, up to invasive carcinoma, resembles the adenoma-carcinoma sequence of the large bowels. PMID- 11382318 TI - Rationale for endoscopic management of adenoma of the papilla of Vater: options and limitations. AB - BACKGROUND: Several studies and our own results prove that endoscopic therapy in selected cases of benign adenomas is safe and technically feasible. In patients refusing surgery or patients with high comorbidity and poor physical health status, endoscopic resection is an excellent alternative. DISCUSSION: The decision for endoscopic or surgical excision of adenomas is determined by general health status, histology, size, location, and depth of the lesion. In carcinoma of the papilla of Vater it is important to assess the tumoral ductal infiltration correctly to determine whether endoscopic resection is a viable option. Intraductal ultrasound is essential before initiating treatment and it therefore contributes to conservative therapy in patients with tumors of the papilla of Vater. Temporary placement of a short pancreatic duct stent may protect against pancreatitis and might allow more excessive ablation of adenomatous tissue, especially around the pancreatic duct orifice. After endoscopic sphincterotomy, biliary and pancreatic endoprostheses can be inserted easily in cases of obstructed pathways or cholangitis and pancreatitis due to tumor obstruction. Argon plasma coagulation can be used to treat oozing tumor hemorrhages or to vaporize tumoral residues after endoscopic snare resection. Endoscopic surveillance is essential after surgical or endoscopic resection of adenomas of the papilla of Vater. PMID- 11382319 TI - Morphologic changes of the anal sphincter musculature during and after temporary stool deviation. AB - BACKGROUND AND AIMS: Temporary stool deviation, using a stoma, is a well-known surgical principle to protect low colorectal or coloanal anastomoses. The purpose of this study was to evaluate any morphologic changes with regard to the anal sphincter muscles during and after temporary ileostomy. PATIENTS AND METHODS: Forty-four patients with rectal carcinomas were studied prospectively. All patients underwent low anterior resection. Reconstruction was performed using either a coloanal pouch or a straight end-to-end anastomosis. A protective stoma was fashioned in all 44 patients (ileostomy n=41; colostomy n=3). Stoma closure was carried out after a median of 85 days (41-330 days). Using a standard protocol, anal-sphincter thickness [m. puborectalis, external anal sphincter (EAS) and internal anal (IAS) sphincter] was assessed by means of endoanal ultrasonography preoperatively, at the time of stoma closure, and every 3 months thereafter for 1 year. RESULTS: The diameter of the puborectal muscle decreased from a median preoperative value of 6.3 mm to 5.7 mm at the time of stoma closure (P=0.03). After 3 months, 6.2 mm was measured. This value remained stable for the complete follow-up period. Similar results were recorded for the EAS. The IAS thickness remained stable throughout the study period, measuring between 2.1 mm and 2.4 mm. CONCLUSION: Temporary stool deviation does lead to morphologic changes of the anal sphincter. While the smooth muscle remains unchanged, the striated counterpart undergoes atrophic transformation. However, after passage reconstruction, i.e., stoma closure, a rapid regeneration of the voluntary muscles is observed. PMID- 11382320 TI - The value of posterior levator repair in the treatment of anorectal incontinence due to rectal prolapse--a clinical and manometric study. AB - BACKGROUND: When treating a complete rectal prolapse, the most important objective is elimination of the prolapse. In addition, restoration of sufficient anorectal continence is extremely important for the patients. We examined the value of posterior levator repair with respect to stabilization of the pelvic floor and to improvement in anorectal incontinence. METHODS: In patients with disabling anorectal incontinence, a posterior levatorplasty can be concomitantly performed during operative removal of the prolapse. To facilitate evaluation of the operative results, we implemented a scoring system to judge the patients' subjective symptoms of incontinence; in addition, we performed manometric measurements of resting and squeezing pressures of the anal sphincter to objectify the anorectal incontinence. RESULTS: From 1991 to 1997, 84 patients (mean age 65+/-10 years, 38-91 years; 79 women, 5 men) with complete rectal prolapse and severe incontinence were operatively treated; corresponding follow ups were done. The following procedures were performed: Frykmann-Goldberg, 28 patients; Wells, 18 patients; Ripstein, 22 patients; and perineal proctectomy, 16 patients. Incontinence for liquid and solid stools was present in all of these patients. Posterior levatorplasty was implemented in 38 patients, and in this group we found significantly better postoperative results, both clinically and in the manometric measurements. Continence was improved by 84% in the group with levatorplasty, but improvement was only 67% in the other group (P<0.05). The incontinence score decreased significantly in the group with levatorplasty (preoperative 16.4+/-3.1, postoperative 9.3+/-4.5, P<0.05, vs the other group with preoperative 15.6+/-4.2, postoperative 11.5+/-5.1). Manometric observations in the group with levatorplasty demonstrated 55% improvement in resting pressure (preoperative 29+/-17 cm H2O, postoperative 45+/-21 cm H2O, P<0.05) and 40% improvement in squeezing pressure (preoperative 61+/-25 cm H2O, postoperative 85+/-31 cm H2O, P<0.05). In the group without levatorplasty, resting and squeezing pressure improved only by 20% (resting pressure: preoperative 32+/-16 cm H2O, postoperative 38+/-18 cm H2O; squeezing pressure: preoperative 64+/-29 cm H2O, postoperative 75+/-26 cm H2O). CONCLUSIONS: Posterior levatorplasty is an easy and efficient operative procedure which facilitates an improvement in anorectal continence. There are no apparent disadvantages. For this reason, levatorplasty can be part of operative procedures implemented in the treatment of a complete rectal prolapse accompanied by disabling anorectal incontinence. PMID- 11382321 TI - Comparison of colonic J-pouch reconstruction and straight coloanal anastomosis after intersphincteric rectal resection. AB - The tendency towards sphincter-preserving resection for distal rectal cancers has led to the technique of straight coloanal anastomosis (CAA) and colonic J-pouch anal anastomosis (CPA) after low anterior resection. The aim of the present study was to compare complication rate, anorectal physiology and functional results after both types of reconstruction after ultra-low intersphincteric resection. A total of 31 patients who had undergone CPA were followed up prospectively using anorectal manometry and a standardised questionnaire and were compared with 63 patients who had undergone CAA and were followed up in the same way. The complication rate after CPA did not differ significantly from that after CAA. One year postoperatively, the median stool frequency and urgency were reduced after CPA (1.7+/-2.2/day; 7% vs. 2.4+/-3.6/day; 14%; P<0.05). Three months after colostomy/ileostomy closure, the maximum tolerable volume, threshold volume and compliance were decreased after CAA when compared with CPA (55+/-12, 34+/-12, and 3.9+/-0.3 ml/mmHg vs. 85+/-21, 53+/-11 and 6.2 ml/mmHg, respectively; P<0.05). Anal manometry revealed no significant differences in the anal resting and squeeze pressure. One year postoperatively, continence also did not differ significantly between CPA and CAA. Colonic J-pouch reconstruction seems to be superior to the straight coloanal anastomosis, especially during the first postoperative year. In view of the often poor prognosis of the patients, it is the reconstruction of choice after ultra-low resections of the rectum. PMID- 11382322 TI - Technique and results of laparoscopic adrenalectomy. AB - The aim of this report is to evaluate the benefits of laparoscopic adrenalectomy in terms of perioperative morbidity, complications and patients recuperation. We reviewed our experience with laparoscopic adrenalectomy in 47 consecutive patients who underwent adrenalectomy over a 4-year period. We used the lateral transperitoneal approach in all cases. The indications for adrenalectomy were Conn's adenoma in 24 patients, pheochromocytoma in 11, Cushing's syndrome in 3 and incidental adrenal tumour in 9. The average duration of surgery was 130 min (range, 60-300 min) and average adrenal gland size was 3.4 cm (range, 1.2-8 cm). Conversion from laparoscopy to laparotomy was necessary in three patients (6.4%), and postoperative complications occurred in two patients. There was no mortality. Laparoscopic adrenalectomy can be considered the method of choice for managing almost all adrenal masses, because of its low morbidity and short postoperative recovery. The main difficulty is to identify the adrenal gland, so several technical procedures are suggested. PMID- 11382323 TI - Intraperitoneal capsaicin treatment reduces postoperative gastric ileus in awake rats. AB - INTRODUCTION: Postoperative gastric ileus interferes with postoperative recovery of the patients. Previous studies suggest that capsaicin-sensitive afferent neurons are involved in the mediation of postoperative gastric ileus. METHODS: A group of rats were equipped with a strain gauge transducer sutured to the gastric wall. Gastric motility was recorded after intraperitoneal injection of capsaicin (0.1 micromol/kg and 1 micromol/kg) or vehicle. The rats were given 2 days of recovery before gastric motility was investigated in a postoperative ileus model. RESULTS: Pretreatment with capsaicin 2 days prior to abdominal surgery significantly increased postoperative gastric motility, with complete recovery of gastric motility at 30 min postoperatively (with the baseline motility index set at 100+/-4%, the gastric motility index 30-45 min postoperatively was 64+/-4% for the vehicle, 138+/-20% for capsaicin 0.1 micromol/kg, and 110+/-12% for capsaicin 1 micromol/kg: P=0.0008 vehicle vs capsaicin). In contrast, capsaicin treatment 2 h prior to abdominal surgery did not increase postoperative gastric motility (gastric motility index 30-45 min postoperatively was 64+/-4% for the vehicle and 51+/-8% for capsaicin 0.1 micromol/kg). The acute intraperitoneal injection of capsaicin decreased gastric motility by about 50-60%, the response lasting for 15 30 min. CONCLUSIONS: Intraperitoneal capsaicin treatment 2 days prior to abdominal surgery resulted in immediate recovery of postoperative gastric motility, indicating an important role for serosal visceral afferent nerve fibers in the mediation of postoperative gastric ileus. Possibly, capsaicin or vanilloid (capsaicin) receptor agonists can be used to treat postoperative ileus in the future. PMID- 11382324 TI - Ultrasound-guided percutaneous cholecystostomy in high-risk surgical patients. AB - BACKGROUND AND AIMS: In critically ill patients, cholecystectomy is associated with a high mortality rate. The aim of this study was to evaluate the safety, efficacy and long-term outcome of ultrasound-guided percutaneous cholecystostomy (USGPC) in critically ill patients with acute cholecystitis. MATERIALS AND METHODS: Clinical records of 51 patients, all considered high-risk surgical patients, with acute cholecystitis treated with USGPC between 1987 and 1999, were retrospectively reviewed. Response was defined as improvement in clinical symptoms and signs, and/or reduction in c-reactive protein and white blood count levels within 72 h. Long-term results were evaluated by means of clinical records and written correspondence. RESULTS: Gallbladder stones were seen in 28 patients whereas 23 had acalculous cholecystitis. Ninety percent showed clinical improvement after USGPC. Cholecystectomy was performed in 16%, of which 6% after recurrent cholecystitis. Recurrence of cholecystitis occurred in 22%. Hospital mortality was 16%. None of the deaths was procedure related or related to acute cholecystitis alone. Major complications relating to the USGPC were rare (4%), while minor catheter-related complications were quite common. CONCLUSIONS: USGPC is a procedure with few complications and a high success rate. In patients with acalculous cholecystitis as well as in many patients with calculous cholecystitis, no further treatment was needed. PMID- 11382325 TI - Influence of suture technique on laparotomy wound healing: an experimental study in the rat. AB - A suture length to wound length ratio (SLWL ratio) of 4:1 for laparotomy closure has proven in clinical studies to reduce incisional hernia incidence. The effect of different SLWL ratios on the mechanical qualities of the healing incision has not been examined experimentally. In 50 rats, the musculo-fascial layer of median laparotomies was closed with polypropylene sutures using SLWL ratios of 8:1, 4:1, 2:1 and 1.7:1. Single and running sutures, different tissue bites and different suture tensions were applied. Five rats served as controls. After 14 days, the horizontal strength of the incision was tested in a digitised tensiometer. The SLWL ratio, suture tension and suture technique proved to have significant influences on the mechanical strength of the incision. Running sutures and especially closures with a ratio of between 4:1 and 8:1 proved significantly stronger than wounds closed with single sutures. When small tissue bites were applied, the positive influence of running sutures was equalised in the early phase of wound healing. High suture tension led to significantly weaker scars independent of the applied suture technique. In accordance with clinical data, it could be proven experimentally that running closure of midline laparotomies with a SLWL ratio above 4:1 avoiding high suture tension exerts a significantly positive effect on the mechanical strength of the incision. Further studies are needed to allow measurement and better control of suture tension. PMID- 11382326 TI - Carcinogenesis of malignant lesions of the gall bladder. The impact of chronic inflammation and gallstones. AB - Gallbladder carcinoma is an uncommon but highly malignant tumor with a poor 5 year survival rate. The presence of gallstones is a well-established risk factor for gallbladder carcinoma, and the risk seems to correlate with stone size. Metaplastic changes of the gallbladder epithelium present in chronic cholecystitis may be a premalignant lesion. Solitary polyps with a size of greater than 1 cm are recognized as a predisposing factor for gallbladder carcinoma when their characteristics are echopenic, sessile, and high cell density. Endoscopic ultrasound is the most useful technique to detect the early changes of malignancy in polyps. Anomalous junction of pancreaticobiliary ducts (AJPBD) without a choledochal cyst and porcelain gallbladder is an additional risk factor for gallbladder malignancy. At the molecular level, it has been proposed that chronic inflammation of the gallbladder may lead to the loss of p53 gene heterozygosity and excessive expression of p53 protein. Furthermore, a proposed mechanism underlying the high risk of gallbladder carcinoma in patients with AJPBD is that chronic reflux of pancreatic juice causes intestinal metaplasia, hyperplasia, and dysplasia with the mutation of p53 and K-ras. In contrast, the causal relationship between porcelain gallbladder and malignancy is yet to be established. In this article, recognition of risk factors for gallbladder carcinoma was summarized with special attention to gallstones and chronic inflammation. PMID- 11382327 TI - Laparoscopic surgery of the spleen: state of the art. AB - INTRODUCTION: Laparoscopic splenectomy (LS) offers superior visualization and access to the spleen and avoids the major laparotomy incision necessary in open splenectomy (OS). This review summarizes the current knowledge of laparoscopic techniques for splenectomy from the perspective of surgeons whose combined experience now totals 340 cases. BACKGROUND AND DISCUSSION: While LS has been applied across the spectrum of splenic diseases, it is most indicated in treatment of a benign hematologic condition with a normal or slightly enlarged spleen as seen in autoimmune thrombocytopenic purpura (ITP), autoimmune deficiency syndrome-related ITP, hemolytic anemia, or spherocytosis. Both anterior and lateral approaches have been used for LS. While benefits of the anterior approach include access to the splenic artery along the superior border of the pancreas within the lesser sac, thus securing vascular control early in the procedure, the lateral approach allows for improved exposure of and access to the splenic pedicle. Also, mechanics and sequence of dissection are enhanced and more intuitive to the surgeon using the lateral approach, and the tail of the pancreas is more easily identified. Potential perioperative complications of LS include hemorrhage, injury to the tail of the pancreas, and deep vein thrombosis. The most common criticisms facing LS are the potential for missed accessory spleens, longer operating time, and greater operating room costs compared to OS. However, while LS requires a longer operating time than OS, studies indicate shorter postoperative hospital stays for LS versus OS patients in comparable cases, which can, in turn, reduce the total hospital cost for the procedure. CONCLUSION: Although LS continues to pose certain technical challenges--such as management of the massive spleen, specimen extraction, and identification of remotely located accessory spleens--its advantages over OS in terms of faster postoperative recovery, shorter hospital stay, and equivalent or lower perioperative morbidity are now well established. Indications for LS and more laparoscopic spleen-conserving surgery are likely to broaden. PMID- 11382328 TI - Nursing shortage: not a simple problem - no easy answers. AB - The article examines the issue of nursing workforce planning within the present health system environment and addresses ethical issues related to the shortage. Numerous factors are influencing both the supply of registered nurses as well as the demand for nursing services. Of particular concern is the negative impact that the current nursing practice environment is having on the retention of registered nurses as well as the ability of the profession to recruit students. Other factors driving this present nursing shortage include the increasing age of working nurses and the potential for retirement; the aging of nursing faculty impacting the capacity of nursing schools; and poor wage compensation. The continual swing in the supply of and demand for nursing services will always be present; however, it is time that nursing sat at the policy-making table with the other decision makers to better anticipate the nature of these changes and how to address them. PMID- 11382329 TI - The nursing shortage in California: the public policy role of the California strategic Planning Committee For Nursing. AB - California is experiencing a nursing shortage resulting from multiple factors. The purpose of this article is to summarize nursing shortage issues in California and present the role of the California Strategic Planning Committee for Nursing (CSPCN) in addressing the issues from a public policy perspective. Reasons for the development of CSPCN as well as goals, objectives, and phases of work are presented. Selected findings from CSPCN's 1999 synthesis of data on the demand for and supply of nurses as well as resulting recommendations are summarized. The role of CSPCN in public policy initiatives is described. PMID- 11382330 TI - The nursing shortage: a continuing challenge: the shortage of educationally prepared nursing faculty. AB - The profession is facing a unique shortage of nursing faculty. Schools of Nursing have always had a shortage of doctorally prepared nursing faculty. The academic norm for faculty teaching in baccalaureate and graduate programs to hold earned doctoral degrees was established later in nursing than for other disciplines. At best, 50% of nursing faculty in such programs have met this academic standard. In addition, the shortage is predicted to escalate in the next decade due to the retirement of a "graying professoriate" of nursing faculty. This article outlines a number of factors influencing the shortage of nursing faculty. Several institutional School of Nursing strategies are suggested for countering the shortage situation. In addition, several national professional and policy strategies are suggested. More creative strategies need to be developed and evaluated by Schools of Nursing and by the profession. PMID- 11382331 TI - The nursing shortage: solutions for the short and long term. AB - With predictions that this nursing shortage will be more severe and have a longer duration than has been previously experienced, traditional strategies implemented by employers will have limited success. The aging nursing workforce, low unemployment, and the global nature of this shortage compound the usual factors that contribute to nursing shortages. For sustained change and assurance of an adequate supply of nurses, solutions must be developed in several areas: education, healthcare deliver systems, policy and regulations, and image. This shortage is not solely nursing's issue and requires a collaborative effort among nursing leaders in practice and education, health care executives, government, and the media. This paper poses several ideas of solutions, some already underway in the United States, as a catalyst for readers to initiate local programs. PMID- 11382332 TI - Threshold dose-response in radiation carcinogenesis: an approach from chronic beta-irradiation experiments and a review of non-tumour doses. AB - PURPOSE: To discuss the threshold dose problem in radiation carcinogenesis after a review of the present author's experimental data on mouse tumour induction by chronic beta-irradiation and other relevant data. CONCLUSIONS: A threshold dose response in radiation carcinogenesis appears in certain tissues and under certain conditions. The optimum condition for demonstrating an apparent threshold is with partial-body chronic or repeated radiation rather than with acute whole-body radiation. Its possible mechanism is host tolerance, involving DNA repair, apoptosis and an immune response activated by low radiation doses. This tolerance level was examined by a survey in the literature of non-tumour-inducing doses, D(nt), the highest dose at which no significant increase of tumours was observed above the control level. PMID- 11382333 TI - Time-course of translocation and dicentric frequencies in a radiation accident case. AB - PURPOSE: To assess the persistence of exchange aberrations measured by FISH chromosome painting after accidental radiation exposure. MATERIALS AND METHODS: Chromosome analyses were carried out in peripheral lymphocytes of a 13-year-old boy exposed to protracted low dose-rate whole-body and short-time partial-body irradiation from a radiation accident in Estonia in 1994. Up to November 1998, the frequencies of translocations and dicentrics were periodically measured using FISH chromosome painting of the target chromosomes 1, 4 and 12, with a simultaneous pancentromeric probe. RESULTS: For the yields of dicentrics, an expected rapid temporal decline was found with a half-time of 14.2+/-1.9 months. The yields of reciprocal translocations also revealed a gradual but significant reduction with a half-time of 51.7+/-12.7 months. CONCLUSION: An unchanged temporal persistence of so-called stable translocations cannot be assumed. Any significant reduction of this aberration type with time obviously limits the application of FISH-based translocation measurements for reliable long-term biodosimetry after combined protracted whole-body and partial-body radiation exposure. PMID- 11382334 TI - Alpha hit frequency due to radon decay products in human lung cells. AB - PURPOSE: To calculate the hit probabilities by alpha-particles emitted by radon progeny for basal and secretory cell nuclei in the epithelium of the human tracheobronchial tree. MATERIALS AND METHODS: The equilibrium activities on the surface of airway tubes were calculated using the ICRP66 model. The stopping power and ranges of alpha-particles in tissue were adopted from ICRU49. A semi analytical method for determining the alpha-particle fluence rate in tissue was applied. The distributions of secretory and basal cells throughout the tracheobronchial tree were as given by Mercer et al. (1991). RESULTS: The probability to hit basal cell nuclei is three-to-four times smaller than for secretory cell nuclei in the bronchial region (BB). However, the total number of traversed basal cell nuclei is greater because of the larger volume abundance of basal cells in BB. The total number of secretory cell nuclei hit in the bronchiolar region (bb) is larger than that in BB because the volume abundance of secretory cells in bb is larger than that in BB. CONCLUSIONS: Basal cells are more sensitive to alpha-radiation than secretory cells. This finding is based on the analysis of the relative number of cell hits and the relative frequencies of lung cancer induction in BB and bb. PMID- 11382335 TI - Neuron loss during early adulthood following prenatal low-dose X-irradiation in the mouse brain. AB - PURPOSE: Apart from subsequent cell death, little is known about long-term effects of a prenatal low-dose X-irradiation (PLDI) on nuclear (n) and mitochondrial (mt) DNA, and whether these effects are connected with reduced neuron numbers in the adult brain. MATERIALS AND METHODS: Pregnant mice were X irradiated with 0, 10 or 50cGy at day 13 (E13) of pregnancy. One day after (E14), or postnatally at day 25 (P25) or P180, the brains of the offspring were analysed concerning the extent of nDNA repair, mt biogenesis, and the relative content of nDNA single strand breaks (SSB). Stereology was applied for evaluating neuronal loss. RESULTS: One day after irradiation no unrepaired SSB were detected. Significant results were mainly obtained for hippocampal pyramidal cells at P180, particularly cell loss following 50 cGy PLDI, increased SSB content and mt biogenesis (0 vs. 10cGy) but decreased mt biogenesis for 10 vs. 50 cGy. CONCLUSIONS: A hypothesis closely related to that regarding molecular events during aging is presented for explaining this second wave of cell death in adult mice following PLDI as a result of accumulated mtDNA damage caused by PLDI. A possible relation to the neurodegenerative hypothesis of schizophrenia is discussed. PMID- 11382336 TI - Effect of irradiation at the early foetal stage on adult brain function of mouse: learning and memory. AB - PURPOSE: To study the long-term effect of early foetal irradiation on the learning and memory in the adult mouse. MATERIALS AND METHODS: The abdominal area of pregnant Swiss albino mice was exposed to a single dose of 0.25-1.5Gy gamma radiation on the 14th day of gestation and the mice were left to deliver their offspring. At 6 months of age, the learning and memory functions of the F(1) mice were tested by hole-board activity, conditioned avoidance response and radial arm maze performance. The animals were again subjected to the radial arm maze test at 12 and 18 months of age. RESULTS: There was a significant dose-dependent decrease in the learning ability and memory retention of 6-month-old mice at doses > 0.25Gy. The significant changes persisted to 18 months of age in mice exposed to >or= 0.5Gy. All changes showed a linear dose-response at doses < 1Gy. CONCLUSIONS: The gestational day 14 of Swiss albino mice is a sensitive stage in brain development to gamma-ray-induced impairment of learning and memory during the adult life. Permanent deficits in these functions can be induced by a dose of approximately 0.5Gy at this stage, when the developmental activity of the cerebral cortex is at its peak. PMID- 11382337 TI - Influence of continuous, very low-dose gamma-irradiation on the mouse immune system. AB - PURPOSE: To investigate whether continuous, very low-dose gamma-irradiation (10 cGy/year) modifies immune parameters in mice. MATERIAL AND METHODS: C57BL/6 female mice, 4 weeks old, were irradiated for 24 months and compared with control mice living in the same room. B- and T-cell subsets were evaluated by flow cytometry before and after stimulation with lectins; subclasses of immunoglobulins were determined by ELISA 2, 4, 6, 8, 12, 18 and 24 months after the beginning of the irradiation. RESULTS: No difference was found in the percentage of CD4(+) and CD8(+) cells in the thymus and the spleen, or in the reactivity of T-cells to lectins. While the number of B-cells in the spleen remained unchanged, a significant decrease of IgG1, IgG2b and IgG2a was observed after respectively 12, 18 and 24 months of irradiation. CONCLUSION: The parameters of cellular immunity studied were not affected by this chronic low dose of irradiation, but this dose rate is probably too low to induce the hormetic effect previously described. Further investigations are necessary to assess whether the decline of immunoglobulin secretion is indicative of a lower rate of infectious diseases or a defect in B-cell function. PMID- 11382338 TI - RPA facilitates rejoining of DNA double-strand breaks in an in vitro assay utilizing genomic DNA as substrate. AB - PURPOSE: Replication protein-A (RPA) is a heterotrimeric single-stranded DNA binding protein playing essential roles in many aspects of nucleic acid metabolism, including DNA replication, nucleotide excision repair and homologous recombination. Here, the role of RPA in the rejoining of radiation-induced DNA double-strand breaks (DSB) by non-homologous end-joining (NHEJ) was investigated. METHODS AND MATERIALS: A previously described in vitro assay for DSB rejoining was employed. The assay used 'naked' genomic DNA prepared from agarose-embedded G(1)-phase A549 cells as a substrate and extracts prepared from HeLa cells as a source of enzymes. Rejoining of DSB in this assay is absolutely dependent on cell extract and proceeds, under optimal reaction conditions, to an extent similar to that observed in intact cells. For experiments, extracts were supplemented with excess purified recombinant RPA. Alternatively, RPA was removed from the extracts either by fractionation or immunodepletion. RESULTS: Although the rejoining of DSB in vitro was not absolutely dependent on RPA, it proceeded faster and to higher levels of completion when recombinant protein was added to the extracts. Depletion of RPA from extracts reduced the rejoining half-times and addition of purified recombinant protein restored the kinetics of DSB rejoining. Extract fractionation indicated the operation of at least two pathways in DSB rejoining, only one of which was facilitated by RPA. CONCLUSIONS: The results suggest that in addition to its role in homologous recombination, RPA may also have a supportive role in some forms of non-homologous end-joining. PMID- 11382339 TI - Suppression of the radiation-sensitive phenotype of hamster irs1 and irs2 strains selected for resistance to 3-aminobenzamide. AB - PURPOSE: The radiosensitive hamster cell lines irs1 and irs2 have phenotypic similarities to cells defective in the early response to DNA-damaging agents as a result of mutations of the genes encoding poly(ADP-ribose)polymerase (PARP) or ataxia-telangiectasia mutated (ATM). Whether modification of PARP activity through selection of strains resistant to 3-aminobenzamide (3-AB) would affect the radiosensitive phenotype of irs1 and irs2 was examined. MATERIALS AND METHODS: 3-AB-resistant strains of irs1, irs2 and their parent line V79 were established and their sensitivity to DNA-damaging agents was measured. In some 3 AB-resistant strains, the radiation resistance of DNA synthesis and the induction of apoptosis were also assayed. Additionally, a number of aspects of PARP function were measured. RESULTS: Independently selected 3-AB-resistant strains of irs2 showed nearly complete suppression of radiation sensitivity, sensitivity to topoisomerase inhibitors, and radioresistant DNA synthesis. 3-AB-resistant strains of irs1 showed partial suppression of phenotype while 3-AB-resistant strains of V79 had no sensitivity changes. The induction of apoptosis in 3-AB resistant strains of irs2 required substantially higher radiation doses than for irs2 itself. 3-AB-resistant strains had no detectable alteration of PARP level or cleavage following ionizing irradiation and there were no mutations in the PARP gene. CONCLUSIONS: Suppression of radiosensitivity associated with 3-AB resistance has important implications for mechanisms of tolerance to damage because it is able to override responses associated with specific genetic defects. PMID- 11382340 TI - Mechanism of protection against radiation-induced DNA damage in plasmid pBR322 by caffeine. AB - PURPOSE: Caffeine (1,3,7-trimethyl xanthine), a dietary component, has been shown to have widely varying effects on DNA damage induced by UV and ionizing radiation, depending upon pre- or post-irradiation administration and its concentration. Caffeine administered post-UV irradiation is known to inhibit enzymatic repair of DNA lesions, leading to potentiation of damage, whereas its presence before or during irradiation elicits protection in a wide range of test systems: bacteria, cultured human cells, plant seeds and mouse. The purpose of this study is to test whether caffeine present during gamma-irradiation of plasmid DNA, a system devoid of replication and repair, could elicit protection by scavenging free radicals. MATERIALS AND METHODS: Plasmid pBR322 DNA was exposed to gamma-radiation in the presence or absence of caffeine at a dose-rate of 1.20 Gy min(-1) and damage measured as single-strand breaks. To understand the mechanisms of the observed protection, especially under oxic conditions, reaction of caffeine with superoxide radical (O(2)(-)), hydrogen peroxide (H(2)O(2)) and the deoxyribose peroxyl radical (ROO(*)) were studied. RESULTS: Irradiation of pBR322 was observed to induce a dose-dependent increase in single-strand breaks. Caffeine itself did not induce strand breaks but reduced radiation-induced strand breaks at micromolar to millimolar concentrations. Caffeine has been shown to react with the radiation-derived oxidants. The reaction rate constants observed were 7.5x10(1) M(-1) s(-1) with O(2)(-) 1.05x10(8) M(-1) s(-1) with ROO(*) and 8.8x10(1) M(-1) s(-1) with H(2)O(2). CONCLUSIONS: Caffeine effectively protects DNA against ionizing radiation in a system devoid of repair and replication machinery. Thus, DNA protection shown by caffeine is possibly due to the scavenging of radiation-derived primary as well as secondary reactive oxygen species, and this physicochemical protective pathway possibly pre-empts any subsequent inhibitory effect of caffeine on the enzymatic repair of DNA. PMID- 11382341 TI - Chemical modification of 5-[125I]iodo-2'-deoxyuridine toxicity in mammalian cells in vitro. AB - PURPOSE: To address the cytotoxic effects of DNA-incorporated (125)I in Chinese hamster V79 lung fibroblasts under various scavenging conditions. METHODS: The toxic effects of DNA-incorporated 5-[(125)I]iodo-2'-deoxyuridine ((125)IdUrd) were assessed by the colony-forming assay with cells incubated in medium containing serum and/or dimethyl sulphoxide (DMSO). Experiments were carried out at 0.3 or -135 degrees C. RESULTS: When (125)I decays were accumulated at 0.3 degrees C in 10% serum 0, 5 or 10% DMSO, no radioprotection was afforded by 5% DMSO, while the dose modification factor (DMF) for 10% DMSO was 2.0. For cells accumulating decays at 135 degrees C in the presence of 5 or 10% serum, DMSO was radioprotective (DMF= 1.8-1.9). D(0) obtained at each serum concentration correlated strongly (R=0.999) with the scavenging capacity of DMSO. Under these experimental conditions, 10% serum is approximately 3.6 times more protective than 5% serum. CONCLUSIONS: The contribution of indirect mechanisms to the toxicity of (125)I decaying within mammalian cell nuclear DNA can be demonstrated not only with DMSO, but also with the hydroxy radical scavengers present in serum. PMID- 11382342 TI - Micronucleus assay in vivo provides significant prognostic information in human cervical carcinoma; the updated analysis. AB - PURPOSE: Reanalysis after a 5-year follow-up previously presented relationships between spontaneous and radiation-induced micronucleus frequencies in tumour cells and the clinical outcome of patients with advanced stages (II B-IV B) of cervix carcinomas treated with radiotherapy. MATERIALS AND METHODS: Spontaneous and induced in vitro and in vivo micronucleus frequencies were determined and related to clinical parameters. Data were analysed by the univariate Kaplan-Meier method and multivariate Cox proportional hazards model. RESULTS: In univariate analysis stage, spontaneous micronucleus frequency before radiotherapy (MNSP) and per cent increment of micronucleus level in vivo after 20 Gy in relation to spontaneous pretreatment level were statistically significant predictors of 5 year recurrence-free, disease-free and overall survival. Neither micronucleus frequency (MN/BNC at 2 Gy) nor proliferating fraction (%BNC at 0 Gy) estimated in vitro (in primary culture) were related to radiotherapy outcome. The age of patients was not associated with clinical results. Multivariate analysis demonstrated that the clinical stage of disease, the high frequency of spontaneous micronuclei and low-induced micronucleus frequency were independent and significantly unfavourable predictive factors for disease-free and overall survival. But for local control, only high MNSP and low-induced MN frequency were significant negative predictive variables. CONCLUSIONS: A high frequency of micronuclei before radiotherapy and a slight increase of micronucleus frequency during radiotherapy measured after 10 fractions of 2 Gy were independent on stage, statistically significant adverse predictors of clinical outcome in cervical carcinoma patients treated with radiotherapy. PMID- 11382343 TI - Generation and identification of the transient intermediates of allophycocyanin by laser photolytic and pulse radiolytic techniques. AB - PURPOSE: To explore the photophysical and photochemical properties of allophycocyanin (APC) and contribute to the understanding of the molecular mechanism of APC photosensitization. MATERIALS AND METHODS: Laser-flash photolytic and pulse radiolytic techniques were used for the first time to characterize the transient intermediates involved in APC photochemistry and photophysics. The excited triplet state and radical cation of APC were identified by acetone sensitization and one-electron oxidation. RESULTS: The 248-nm laser flash photolysis of APC in N(2)-saturated aqueous solution (pH 7.0) yields the triplet state and radical cation of APC. The APC radical cations were generated by ionization via a monophotonic process, with a quantum yield of 0.17. CONCLUSIONS: APC can undergo both photo-excitation and photo-ionization under the present experimental conditions. These new findings suggest that APC has the potential to act as both a type I and type II photosensitizer. PMID- 11382345 TI - Solar ultraviolet exposures at ground level in tree shade during summer in south east Queensland. AB - Data are presented on the effect of the tree canopy transmittance in the visible waveband (VT), canopy width, height and height of the start of the tree canopy (CH) on the solar UV in tree shade on a horizontal plane at ground level during a Southern Hemisphere summer. Of these factors, the VT and CH have an influence on the UV irradiances in the tree shade. The shade ratios (UV in tree shade to that in full sun) for erythemal UV ranged from 0.71 to 0.42, 0.54 to 0.29 and 0.63 to 0.41 for morning, noon and afternoon, respectively, for the VT range of 0.4-1.0. Over the same VT range, the shade ratios for UVA ranged from 0.61 to 0.28, 0.50 to 0.22 and 0.49 to 0.29 for morning, noon and afternoon, respectively. The UV exposures in the tree shade decreased with the VT with a marginally higher decrease in the irradiances for the UVA compared to the erythemal UV. Despite the protection by the tree shade, significant UV in the tree shade of approximately 4 MED (minimum erythemal dose) were received for the latitude in this research on a cloud free summer day on a horizontal plane over a 2-h period centred about solar noon. PMID- 11382346 TI - Decontamination issues for chemical and biological warfare agents: how clean is clean enough? AB - The objective of this assessment is to determine what level of cleanup will be required to meet regulatory and stakeholder needs in the case of a chemical and/or biological incident at a civilian facility. A literature review for selected, potential chemical and biological warfare agents shows that dose information is often lacking or controversial. Environmental regulatory limits or other industrial health guidelines that could be used to help establish cleanup concentration levels for such agents are generally unavailable or not applicable for a public setting. Although dose information, cleanup criteria, and decontamination protocols all present challenges to effective planning, several decontamination approaches are available. Such approaches should be combined with risk-informed decision making to establish reasonable cleanup goals for protecting health, property, and resources. Key issues during a risk assessment are to determine exactly what constitutes a safety hazard and whether decontamination is necessary or not for a particular scenario. An important conclusion is that cleanup criteria are site dependent and stakeholder specific. The results of a modeling exercise for two outdoor scenarios are presented to reinforce this conclusion. Public perception of risk to health, public acceptance of recommendations based on scientific criteria, political support, time constraints, and economic concerns must all be addressed in the context of a specific scenario to yield effective and acceptable decontamination. PMID- 11382347 TI - Parametric study of selective removal of atmospheric aerosol by coagulation, condensation and gravitational settling. AB - This work studies the scavenging efficiencies of aerosol particles after a given mechanism of removal (coagulation, heterogeneous nucleation and gravitational settling) as a function of time. It also analyses the health impact of the aerosol before and after the above dynamic mechanisms by comparing the respirable dust fractions. The well-known equations of scavenging are applied to three atmospheric environments (clear, hazy and urban) that represent the aerosol PSDs in the countryside, industry and the city, respectively. From this study it is inferred that respirable dust is hardly scavenged and when compared with the initial volume of respirable aerosol, roughly 10% remains after 18 h of gravitational settling. Therefore, gravitational settling is the main removal mechanism of respirable aerosol compared to condensation and coagulation and is almost six times better than rainout. PMID- 11382348 TI - A review of the science behind drinking water standards for copper. AB - Copper is an essential element, being a vital component in several enzyme systems. Some intake, therefore, is necessary for human health. At high intakes, however, it can have toxic effects. This paper uses a quality audit framework to review the strength of the science underlying the setting of standards for copper in drinking water intended for human consumption. The two copper standards examined are the revised EU drinking water directive (98/83/EC) of 2 mg/l, based on WHO guidelines, and the USEPA treatment technique action level of 1.3 mg/l. Examination of these standards reveals that neither has a firm scientific basis, accordingly both are difficult to justify from a scientific standpoint, a situation that is worrying in both health and policy terms. PMID- 11382349 TI - The efficacy of subsurface flow reed bed treatment in the removal of Campylobacter spp., faecal coliforms and Escherichia coli from poultry litter. AB - The use of poultry waste as a fertiliser on arable land is an accepted method of waste treatment. However, run-off from such practices may result in contamination of the watercourse by human pathogens. In this study the effectiveness of using constructed wetlands as an alternative treatment for poultry manure waste was evaluated. Enumeration of Campylobacter spp., Escherichia coli, total coliforms and total aerobes were carried out on influent and effluent samples from reed beds loaded with poultry waste. For both sequential loading and continuous loading there was a statistically significant mean log reduction of 3.56 and 4.25 for E. coli, 3.2 and 3.88 for coliforms, 3.85 and 4.2 for total aerobic counts and 3.13 and 2.96 for Campylobacter spp., respectively. This method, which has been previously recognised as cost-effective and environmentally acceptable, provides an efficient method for reducing numbers of these bacteria in poultry waste and therefore an effective alternative treatment for such waste or waters containing run off from land previously spread with poultry manure. PMID- 11382350 TI - Occupational lead exposure and amino acid profiles and liver function tests in industrial workers. AB - The aim of the present study is to determine the effect of blood lead on the plasma levels of amino acids and serum liver enzymes in industrial workers in United Arab Emirates (UAE). This comparison study consisted of 100 industrial workers (exposed) and 100 non-industrial workers (non-exposed), matched for age, sex and nationality selected from Al-Ain, Abu-Dhabi Emirates. Industrial workers had higher proportion (19%) of smokers than non-industrial workers (11%) which was not considered to be statistically significantly different. Industrial workers had significantly higher mean of blood lead level (77.5 +/- 42.8 miccrog x dl(-1)) than non-industrial workers (19.8 +/- 12.3 microg x dl(-1)). The amino acid analysis showed higher values among industrial than non-industrial workers for histidine, isoleucine, leucine, threonine, lysine, valine, methionine and arginine (essential amino acids, (p<0.0001). Ornithine, taurine, glutamic acid, serine, glycine, proline and alanine (non-essential amino acids) showed significantly higher values in industrial when compared with the non-industrial workers (p<0.0001). Plasma liver function test, cardiac enzymes and renal function test were carried out on industrial and non-industrial workers. The results revealed alkaline phosphatase (p=0.012) and lactate dehydrogenase (p=0.029) were significantly higher in industrial than in non-industrial workers. On the basis of this study, it can be concluded that a substantial difference in amino acid profiles, blood lead and LFT between exposed and non-exposed was found. These results might be related to lead exposure and might have affects on the kidneys or liver. PMID- 11382351 TI - Fuel oil effect on the population growth, species diversity and chlorophyll (a) content of freshwater microalgae. AB - Fresh water algae were subjected to different concentrations (0.03, 0.07, 0.12, 0.25 and 0.5 g x l(-1)) of aqueous extract of reference fuel oil (EPA, USA, API Oil No. 2, 38% aromatic, 1274). Significant decrease in Chlorophyll. (a) was observed as the concentration of fuel oil was increased. The EC50 value of fuel oil after 7 days was 0.29 g x l(-1). Total algal counts and growth rate decreased in response to the studied fuel oil. High diversity values in diatoms were observed in all treated aqueous cultures. High concentrations of fuel oil significantly decreased carbohydrate and protein contents of algal cells. PMID- 11382353 TI - Theoretical analysis of mutation-adaptive evolutionary algorithms. AB - Adaptive evolutionary algorithms require a more sophisticated modeling than their static-parameter counterparts. Taking into account the current population is not enough when implementing parameter-adaptation rules based on success rates (evolution strategies) or on premature convergence (genetic algorithms). Instead of Markov chains, we use random systems with complete connections - accounting for a complete, rather than recent, history of the algorithm's evolution. Under the new paradigm, we analyze the convergence of several mutation-adaptive algorithms: a binary genetic algorithm, the 1/5 success rule evolution strategy, a continuous, respectively a dynamic (1+1) evolutionary algorithm. PMID- 11382354 TI - Convergence in evolutionary programs with self-adaptation. AB - Evolutionary programs are capable of finding good solutions to difficult optimization problems. Previous analysis of their convergence properties has normally assumed the strategy parameters are kept constant, although in practice these parameters are dynamically altered. In this paper, we propose a modified version of the 1/5-success rule for self-adaptation in evolution strategies (ES). Formal proofs of the long-term behavior produced by our self-adaptation method are included. Both elitist and non-elitist ES variants are analyzed. Preliminary tests indicate an ES with our modified self-adaptation method compares favorably to both a non-adapted ES and a 1/5-success rule adapted ES. PMID- 11382355 TI - Completely derandomized self-adaptation in evolution strategies. AB - This paper puts forward two useful methods for self-adaptation of the mutation distribution - the concepts of derandomization and cumulation. Principle shortcomings of the concept of mutative strategy parameter control and two levels of derandomization are reviewed. Basic demands on the self-adaptation of arbitrary (normal) mutation distributions are developed. Applying arbitrary, normal mutation distributions is equivalent to applying a general, linear problem encoding. The underlying objective of mutative strategy parameter control is roughly to favor previously selected mutation steps in the future. If this objective is pursued rigorously, a completely derandomized self-adaptation scheme results, which adapts arbitrary normal mutation distributions. This scheme, called covariance matrix adaptation (CMA), meets the previously stated demands. It can still be considerably improved by cumulation - utilizing an evolution path rather than single search steps. Simulations on various test functions reveal local and global search properties of the evolution strategy with and without covariance matrix adaptation. Their performances are comparable only on perfectly scaled functions. On badly scaled, non-separable functions usually a speed up factor of several orders of magnitude is observed. On moderately mis-scaled functions a speed up factor of three to ten can be expected. PMID- 11382356 TI - Self-adaptive genetic algorithms with simulated binary crossover. AB - Self-adaptation is an essential feature of natural evolution. However, in the context of function optimization, self-adaptation features of evolutionary search algorithms have been explored mainly with evolution strategy (ES) and evolutionary programming (EP). In this paper, we demonstrate the self-adaptive feature of real-parameter genetic algorithms (GAs) using a simulated binary crossover (SBX) operator and without any mutation operator. The connection between the working of self-adaptive ESs and real-parameter GAs with the SBX operator is also discussed. Thereafter, the self-adaptive behavior of real parameter GAs is demonstrated on a number of test problems commonly used in the ES literature. The remarkable similarity in the working principle of real parameter GAs and self-adaptive ESs shown in this study suggests the need for emphasizing further studies on self-adaptive GAs. PMID- 11382357 TI - A comparison study of self-adaptation in evolution strategies and real-coded genetic algorithms. AB - This paper discusses the self-adaptive mechanisms of evolution strategies (ES) and real-coded genetic algorithms (RCGA) for optimization in continuous search spaces. For multi-membered evolution strategies, a self-adaptive mechanism of mutation parameters has been proposed by Schwefel. It introduces parameters such as standard deviations of the normal distribution for mutation into the genetic code and lets them evolve by selection as well as the decision variables. In the RCGA, crossover or recombination is used mainly for search. It utilizes information on several individuals to generate novel search points, and therefore, it can generate offspring adaptively according to the distribution of parents without any adaptive parameters. The present paper discusses characteristics of these two self-adaptive mechanisms through numerical experiments. The self-adaptive characteristics such as translation, enlargement, focusing, and directing of the distribution of children generated by the ES and the RCGA are examined through experiments. PMID- 11382358 TI - Looking beyond selection probabilities: adaptation of the chi(2) measure for the performance analysis of selection methods in GAs. AB - Viewing the selection process in a genetic algorithm as a two-step procedure consisting of the assignment of selection probabilities and the sampling according to this distribution, we employ the chi(2) measure as a tool for the analysis of the stochastic properties of the sampling. We are thereby able to compare different selection schemes even in the case that their probability distributions coincide. Introducing a new sampling algorithm with adjustable accuracy and employing two-level test designs enables us to further reveal the intrinsic correlation structures of well-known sampling algorithms. Our methods apply well to integral methods like tournament selection and can be automated. PMID- 11382360 TI - Modeling amino acid replacement. AB - The estimation of amino acid replacement frequencies during molecular evolution is crucial for many applications in sequence analysis. Score matrices for database search programs or phylogenetic analysis rely on such models of protein evolution. Pioneering work was done by Dayhoff et al. (1978) who formulated a Markov model of evolution and derived the famous PAM score matrices. Her estimation procedure for amino acid exchange frequencies is restricted to pairs of proteins that have a constant and small degree of divergence. Here we present an improved estimator, called the resolvent method, that is not subject to these limitations. This extension of Dayhoff's approach enables us to estimate an amino acid substitution model from alignments of varying degree of divergence. Extensive simulations show the capability of the new estimator to recover accurately the exchange frequencies among amino acids. Based on the SYSTERS database of aligned protein families (Krause and Vingron, 1998) we recompute a series of score matrices. PMID- 11382361 TI - Mutation-tolerant protein identification by mass spectrometry. AB - Database search in tandem mass spectrometry is a powerful tool for protein identification. High-throughput spectral acquisition raises the problem of dealing with genetic variation and peptide modifications within a population of related proteins. A method that cross-correlates and clusters related spectra in large collections of uncharacterized spectra (i.e., from normal and diseased individuals) would be very valuable in functional proteomics. This problem is far from being simple since very similar peptides may have very different spectra. We introduce a new notion of spectral similarity that allows one to identify related spectra even if the corresponding peptides have multiple modifications/mutations. Based on this notion, we developed a new algorithm for mutation-tolerant database search as well as a method for cross-correlating related uncharacterized spectra. PMID- 11382362 TI - Distance-based reconstruction of tree models for oncogenesis. AB - Comparative genomic hybridization (CGH) is a laboratory method to measure gains and losses in the copy number of chromosomal regions in tumor cells. It is hypothesized that certain DNA gains and losses are related to cancer progression and that the patterns of these changes are relevant to the clinical consequences of the cancer. It is therefore of interest to develop models which predict the occurrence of these events, as well as techniques for learning such models from CGH data. We continue our study of the mathematical foundations for inferring a model of tumor progression from a CGH data set that we started in Desper et al. (1999). In that paper, we proposed a class of probabilistic tree models and showed that an algorithm based on maximum-weight branching in a graph correctly infers the topology of the tree, under plausible assumptions. In this paper, we extend that work in the direction of the so-called distance-based trees, in which events are leaves of the tree, in the style of models common in phylogenetics. Then we show how to reconstruct the distance-based trees using tree-fitting algorithms developed by researchers in phylogenetics. The main advantages of the distance-based models are that 1) they represent information about co-occurrences of all pairs of events, instead of just some pairs, 2) they allow quantitative predictions about which events occur early in tumor progression, and 3) they bring into play the extensive methodology and software developed in the context of phylogenetics. We illustrate the distance-based tree method and how it complements the branching tree method, with a CGH data set for renal cancer. PMID- 11382363 TI - Testing for differentially-expressed genes by maximum-likelihood analysis of microarray data. AB - Although two-color fluorescent DNA microarrays are now standard equipment in many molecular biology laboratories, methods for identifying differentially expressed genes in microarray data are still evolving. Here, we report a refined test for differentially expressed genes which does not rely on gene expression ratios but directly compares a series of repeated measurements of the two dye intensities for each gene. This test uses a statistical model to describe multiplicative and additive errors influencing an array experiment, where model parameters are estimated from observed intensities for all genes using the method of maximum likelihood. A generalized likelihood ratio test is performed for each gene to determine whether, under the model, these intensities are significantly different. We use this method to identify significant differences in gene expression among yeast cells growing in galactose-stimulating versus non stimulating conditions and compare our results with current approaches for identifying differentially-expressed genes. The effect of sample size on parameter optimization is also explored, as is the use of the error model to compare the within- and between-slide intensity variation intrinsic to an array experiment. PMID- 11382364 TI - Analysis of variance for gene expression microarray data. AB - Spotted cDNA microarrays are emerging as a powerful and cost-effective tool for large-scale analysis of gene expression. Microarrays can be used to measure the relative quantities of specific mRNAs in two or more tissue samples for thousands of genes simultaneously. While the power of this technology has been recognized, many open questions remain about appropriate analysis of microarray data. One question is how to make valid estimates of the relative expression for genes that are not biased by ancillary sources of variation. Recognizing that there is inherent "noise" in microarray data, how does one estimate the error variation associated with an estimated change in expression, i.e., how does one construct the error bars? We demonstrate that ANOVA methods can be used to normalize microarray data and provide estimates of changes in gene expression that are corrected for potential confounding effects. This approach establishes a framework for the general analysis and interpretation of microarray data. PMID- 11382365 TI - Fidelity of enzymatic ligation for DNA computing. AB - We describe a convenient assay for rapid qualitative evaluation of hybridization/ligation fidelity. The approach uses randomized probe strands of DNA and restriction enzyme digestion after amplification of reaction products by the polymerase chain reaction (PCR). We report ligation efficiencies and fidelities of two DNA ligases, T4 DNA ligase and Thermus aquaticus (Taq) DNA ligase, over a range of temperatures. PMID- 11382366 TI - Representing and reasoning about protein families using generative and discriminative methods. AB - This work addresses the issues of data representation and incorporation of domain knowledge into the design of learning systems for reasoning about protein families. Given the limited expressive capacity of a particular method, a mixture of protein annotation and fold recognition experts, each implementing a different underlying representation, should provide a robust method for assigning sequences to families. These ideas are illustrated using two data-driven learning methods that make use of different prior information and employ independent, yet complementary, projections of a family: hidden Markov models (HMMs) based on a multiple sequence alignment and neural networks (NNs) based on global sequence descriptors of proteins. Examination of seven protein families indicates that combining a generative (HMM) and a discriminative (NN) method is better than either method on its own. Biologically, human 4-hydroxyphenylpyruvic acid dioxygenase, involved in tyrosinemia type 3, is predicted to be structurally and functionally related to the glyoxalase I family. PMID- 11382367 TI - Herpes zoster as an immune reconstitution disease after initiation of combination antiretroviral therapy in patients with human immunodeficiency virus type-1 infection. AB - BACKGROUND: Initiation of combination antiretroviral therapy may be followed by inflammatory reactions. We studied the epidemiology of herpes zoster infection among patients with human immunodeficiency virus (HIV) infection who were treated with combination antiretroviral therapy. SUBJECTS AND METHODS: Of 316 patients who initiated combination antiretroviral therapy, 24 (8%) were treated for herpes zoster within 17 weeks of starting therapy. The characteristics of these cases were compared with those of a control group of 96 HIV-1-infected patients, who were matched by age, sex, plasma HIV-1 RNA concentration and CD4 cell counts, and length of follow-up. RESULTS: The incidence of herpes zoster associated with combination antiretroviral therapy was 9 episodes per 100 patient-years. There were no significant differences between cases and controls in age, sex, years of HIV infection, history of herpes zoster, previous acquired immune deficiency syndrome, or baseline mean CD4 and CD8 cell counts before beginning combination antiretroviral therapy. However, patients who developed herpes zoster had a significantly greater mean (+/- SD) increase in the number of CD8 cells than did controls (347 +/- 269 vs. 54 +/- 331 cells/mL, P = 0.0006). In a multivariate analysis, the only factor that was associated with the development of herpes zoster was the increase in CD8 cells from before initiation of combination antiretroviral therapy to 1 month before development of herpes zoster (odds ratio 1.3 per percentage increase; 95% confidence interval: 1.1 to 1.5; P = 0.0002). CONCLUSION: The initiation of combination antiretroviral therapy in HIV-1 infected patients was often associated with the development of herpes zoster, especially in those in whom the number of CD8 cells increased after therapy. PMID- 11382368 TI - A randomized, controlled trial of interventions to improve adherence to isoniazid therapy to prevent tuberculosis in injection drug users. AB - PURPOSE: To determine the effect of several interventions on adherence to tuberculosis preventive therapy. METHODS: We conducted a randomized trial with a factorial design comparing strategies for improving adherence to isoniazid preventive therapy in 300 injection drug users with reactive tuberculin tests and no evidence of active tuberculosis. Patients were assigned to receive directly observed isoniazid preventive therapy twice weekly (Supervised group, n = 99), daily self-administered isoniazid with peer counseling and education (Peer group, n = 101), or routine care (Routine group, n = 100). Patients within each arm were also randomly assigned to receive an immediate or deferred monthly $10 stipend for maintaining adherence. The endpoints of the trial were completing 6 months of treatment, pill-taking as measured by self-report or observation, isoniazid metabolites present in urine, and bottle opening as determined by electronic monitors in a subset of patients. RESULTS: Completion of therapy was 80% for patients in the Supervised group, 78% in the Peer group, and 79% in the Routine group (P = 0.70). Completion was 83% (125 of 150) among patients receiving immediate incentives versus 75% (112 of 150) among patients with deferred incentives (P = 0.09). The proportion of patients who were observed or reported taking at least 80% of their doses was 82% for the Supervised arm of the study, compared with 71% for the Peer arm and 90% for the Routine arm. The proportion of patients who took 100% of doses was 77% for the Supervised arm (by observation), 6% for the Peer arm (by report), and 10% for the Routine arm (by report; P <0.001). Direct observation showed the median proportion of doses taken by the Supervised group was 100%, while electronic monitoring in a subset of patients showed the Peer group (n = 27) took 57% of prescribed doses and the Routine group (n = 32) took 49% (P <0.001). Patients in the Routine arm overreported adherence by twofold when data from electronic monitoring were used as a gold standard. There were no significant differences in electronically monitored adherence by type of incentive. CONCLUSION: Adherence to isoniazid preventive therapy by injection drug users is best with supervised care. Peer counseling improves adherence over routine care, as measured by electronic monitoring of pill caps, and patients receiving peer counseling more accurately reported their adherence. More widespread use of supervised care could contribute to reductions in tuberculosis rates among drug users and possibly other high-risk groups. PMID- 11382370 TI - Influence of body weight on response to subcutaneous vitamin K administration in over-anticoagulated patients. AB - PURPOSE: To determine the influence of body weight on the international normalized ratio (INR) response to a fixed dose of vitamin K in overanticoagulated patients. METHODS: Retrospective review of records of patients who received 1 mg of vitamin K subcutaneously to correct excessive INR. Dose of vitamin K in milligrams per kilograms plotted against change in INR in 24 hours. RESULTS: Fifteen patients were identified who met all inclusion criteria. Linear regression analysis plotted INR response at 24 hours versus dose of vitamin K adjusted for body weight. Pearson's product moment correlation (R = 0.85) indicated a significant relationship between INR response at 24 hours to an adjusted body weight dose of subcutaneous vitamin K (P = 0.0000523). A strong correlation (r = 0.69) also existed between INR response at 24 hours and the actual body weight dose of subcutaneous vitamin K (P = 0.004). CONCLUSIONS: In overanticoagulated patients, variability in response to vitamin K may be explained by variability in body weight. Dosing vitamin K according to body weight may result in a more predictable INR response. PMID- 11382369 TI - Propionyl-L-carnitine improves exercise performance and functional status in patients with claudication. AB - PURPOSE: We tested the hypothesis that propionyl-L-carnitine would improve peak walking time in patients with claudication. Secondary aims of the study were to evaluate the effects of propionyl-L-carnitine on claudication onset time, functional status, and safety. SUBJECTS AND METHODS: In this double-blind, randomized, placebo-controlled trial, 155 patients with disabling claudication from the United States (n = 72) or Russia (n = 83) received either placebo or propionyl-L-carnitine (2g/day orally) for 6 months. Subjects were evaluated at baseline and 3 and 6 months after randomization with a graded treadmill protocol at a constant speed of 2 miles per hour, beginning at 0% grade, with increments in the grade of 2% every 2 minutes until maximal symptoms of claudication forced cessation of exercise. Questionnaires were used to determine changes in functional status. RESULTS: At baseline, peak walking time was 331 +/- 171 seconds in the placebo group and 331 +/- 187 seconds in the propionyl-L-carnitine group. After 6 months of treatment, subjects randomly assigned to propionyl-L carnitine increased their peak walking time by 162 +/- 222 seconds (a 54% increase) as compared with an improvement of 75 +/- 191 seconds (a 25% increase) for those on placebo (P <0.001). Similar improvements were observed for claudication onset time. Propionyl-L-carnitine treatment significantly improved walking distance and walking speed (by the Walking Impairment Questionnaire), and enhanced physical role functioning, reduced bodily pain, and resulted in a better health transition score (by the Medical Outcome Study SF-36 Questionnaire). The incidence of adverse events and study discontinuations were similar in the two treatment groups. CONCLUSIONS: Propionyl-L-carnitine safely improved treadmill exercise performance and enhanced functional status in patients with claudication. PMID- 11382371 TI - Correlates of hepatitis B vaccination in a high-risk population: an Internet sample. AB - PURPOSE: We sought to identify factors associated with hepatitis B virus vaccination, including knowledge and attitudes about hepatitis vaccination, and sexual and nonsexual risk behaviors among at-risk homosexual and bisexual men. SUBJECTS AND METHODS: Internet electronic communications were used to collect data from homosexual and bisexual men from the United States, using a 31-item online questionnaire accessible for 1 month. RESULTS: The mean (+/- SD) age of the 336 respondents was 38 +/- 11 years. Nearly 42% (142) reported at least one dose of vaccine; the remainder were completely unvaccinated. About 21% (n = 71) reported having no information about hepatitis. Approximately 72% (242) of respondents reported never using condoms during oral intercourse, and 26% (n = 87) reported using condoms during less than half of their episodes of anal intercourse. In multivariate analysis, variables associated with vaccination were younger age (odds ratio [OR] 0.7 per 10-year increase in age; 95% confidence interval [CI] 0.59 to 0.84, P = 0.002), high level of knowledge about the vaccine (OR 1.4; 95% CI: 1.03 to 1.83, P = 0.007), communication with a health-care provider about hepatitis (OR 1.98; 95% CI 1.31 to 2.98, P = 0.006), and professional training that included hepatitis education (OR 2.77; 95% CI 1.7 to 4.5, P = 0.001). CONCLUSIONS: Our findings underscore the need for health care providers to emphasize vaccine efficacy and safety, and to encourage high-risk patients to receive vaccination, particularly among men at high risk based on sexual and drug use behaviors. PMID- 11382372 TI - Noble heart. PMID- 11382373 TI - New heparin dosing recommendations for patients with acute coronary syndromes. AB - Despite major innovations in antithrombotic and antiplatelet therapy, unfractionated intravenous heparin is widely used to treat acute coronary syndromes. Recommendations for unfractionated heparin dosing in acute myocardial infarction and unstable angina have been issued in two recent American College of Cardiology/American Heart Association guidelines. An initial heparin bolus of 60 U/kg (maximum, 4000 U) followed by a 12-U/kg/h infusion (maximum 1000 U/h) is recommended with alteplase for ST-elevation myocardial infarction. When intravenous heparin is administered for myocardial infarction with non-ST elevation and unstable angina, an initial bolus of 60 to 70 U/kg (maximum, 5000 U) followed by a 12- to 15-U/kg/h infusion is recommended. The goal is to achieve an activated partial thromboplastin time of 50 to 70 seconds. Here, we review these new dosing regimens and explain the rationale for their use. We also review the risk of bleeding with heparin, especially when administered concurrently with aspirin, thrombolytic agents, and glycoprotein IIb/IIIa antagonists, and the relationship between activated partial thromboplastin time and cardiac events. PMID- 11382374 TI - Whiplash: a review of a commonly misunderstood injury. AB - Whiplash injury is a relatively common occurrence, but its mechanism and optimal treatment remain poorly understood. It is estimated that the incidence of whiplash injury is approximately 4 per 1,000 persons. The most common radiographic findings include either preexisting degenerative changes or a slight flattening of the normal lordotic curvature of the cervical spine. Computed tomography and magnetic resonance imaging are generally reserved for cases of neurologic deficit, suspected disc or spinal cord damage, fracture, or ligamentous damage. Biomechanics studies have determined that after rear impact C6 is rotated back into extension before movement of the upper cervical vertebrae. Thus, the lower cervical vertebrae were in extension while the upper vertebrae were in a position of relative flexion, producing an S shape in the cervical spine. It is believed that this abnormal motion pattern might play a role in the development of whiplash injuries. Historically, a soft cervical collar has been used early after the injury in an attempt to restrict cervical range of motion and limit the chances of further injury. More recent studies report rest and restriction of motion to be detrimental and to slow the healing process. PMID- 11382375 TI - Patent ductus venosus in an adult presenting as pulmonary hypertension, right sided heart failure, and portosystemic encephalopathy. PMID- 11382378 TI - Human immunodeficiency virus-associated immune reconstitution disease. PMID- 11382379 TI - Expanding directly observed therapy: tuberculosis to human immunodeficiency virus. PMID- 11382380 TI - Whiplash. PMID- 11382381 TI - Why what we do matters. PMID- 11382382 TI - Long-term alteration of calcium homeostatic mechanisms in the pilocarpine model of temporal lobe epilepsy. AB - The pilocarpine model of temporal lobe epilepsy is an animal model that shares many of the clinical and pathophysiological characteristics of temporal lobe or limbic epilepsy in humans. This model of acquired epilepsy produces spontaneous recurrent seizure discharges following an initial brain injury produced by pilocarpine-induced status epilepticus. Understanding the molecular mechanisms mediating these long lasting changes in neuronal excitability would provide an important insight into developing new strategies for the treatment and possible prevention of this condition. Our laboratory has been studying the role of alterations in calcium and calcium-dependent systems in mediating some of the long-term neuroplasticity changes associated with epileptogenesis. In this study, [Ca(2+)](i) imaging fluorescence microscopy was performed on CA1 hippocampal neurons acutely isolated from control and chronically epileptic animals at 1 year after the induction of epileptogenesis with two different fluorescent dyes (Fura 2 and Fura-FF) having high and low affinities for [Ca(2+)](i). The high affinity Ca(2+) indicator Fura-2 was utilized to evaluate [Ca(2+)](i) levels up to 900 nM and the low affinity indicator Fura-FF was employed for evaluating [Ca(2+)](i) levels above this range. Baseline [Ca(2+)](i) levels and the ability to restore resting [Ca(2+)](i) levels after a brief exposure to several glutamate concentrations in control and epileptic neurons were evaluated. Epileptic neurons demonstrated a statistically significantly higher baseline [Ca(2+)](i) level in comparison to age-matched control animals. This alteration in basal [Ca(2+)](i) levels persisted up to 1 year after the induction of epileptogenesis. In addition, the epileptic neurons were unable to rapidly restore [Ca(2+)](i) levels to baseline following the glutamate-induced [Ca(2+)](i) loads. These changes in Ca(2+) regulation were not produced by a single seizure and were not normalized by controlling the seizures in the epileptic animals with anticonvulsant treatment. Peak [Ca(2+)](i) levels in response to different concentrations of glutamate were the same in both epileptic and control neurons. Thus, glutamate produced the same initial [Ca(2+)](i) load in both epileptic and control neurons. Characterization of the viability of acutely isolated neurons from control and epileptic animals utilizing standard techniques to identify apoptotic or necrotic neurons demonstrated that epileptic neurons had no statistically significant difference in viability compared to age-matched controls. These results provide the first direct measurement of [Ca(2+)](i) levels in an intact model of epilepsy and indicate that epileptogenesis in this model produced long-lasting alterations in [Ca(2+)](i) homeostatic mechanisms that persist for up to 1 year after induction of epileptogenesis. These observations suggest that altered [Ca(2+)](i) homeostatic mechanisms may underlie some aspects of the epileptic phenotype and contribute to the persistent neuroplasticity changes associated with epilepsy. PMID- 11382383 TI - Par-4 is a synaptic protein that regulates neurite outgrowth by altering calcium homeostasis and transcription factor AP-1 activation. AB - Although Par-4 (prostate apoptosis response-4) is involved in initiation of neurodegenerative cascades associated with certain neurodegenerative disorders, normal physiological roles of Par-4 in neurons have remained elusive. It was recently reported that Par-4 protein levels could be regulated at translational level in synaptic terminals following apoptotic insults, suggesting that Par-4 might play a role in synaptic function. We report that Par-4 is a synaptic protein preferably localized in postsynaptic density (PSD). The expression of Par 4 in synaptosome preparations and PSDs are developmentally and regionally regulated. Synaptic Par-4 is enriched in the cerebral cortex and the hippocampus, but not in the cerebellum. In vitro as well as in vivo experiments demonstrate that the levels of synaptic Par-4 increase as the neurons mature. Overexpression of Par-4 in transfected PC12 cells inhibits nerve growth factor (NGF)-induced cellular differentiation and neurite outgrowth by a mechanism involving aberrant elevation of intracellular calcium levels and suppression of activation of the transcription factor AP-1. The actions of Par-4 were consistently blocked by co expression of the dominant negative regulator of Par-4 activity (the leucine zipper domain of Par-4). Since the leucine zipper domain of Par-4 (Leu.zip) may mediate protein--protein interactions, the results indicate that the actions of Par-4 require its interaction with other protein(s) or dimerization with itself. These results suggest that Par-4 may play an important role in postsynaptic signal transduction and regulation of cellular pathways associated with cellular differentiation and neurite outgrowth. Identification of Par-4 as a novel synaptic protein may have significant implications in understanding the mechanisms of synaptic functions in physiological and pathological settings. PMID- 11382384 TI - Cellular mRNA expression of the transcription factor NGFI-B suggests a gene regulatory role in striatal opiate-peptide neurons. AB - Previous studies have shown that NGFI-B mRNA is highly expressed in the adult striatum. In the present study we analyzed the anatomical distribution of NGFI-B mRNA within this brain region as well as the degree of co-existence of NGFI-B with different striatal markers in the adult brain. NGFI-B mRNA levels were found to be significantly higher within the dorsomedial portion of the striatum as compared to the ventrolateral striatum. This distribution pattern was maintained throughout the rostro--caudal axis of the striatum. Double in situ hybridization studies showed that striatal NGFI-B mRNA colocalized with a subset of preproenkephalin and prodynorphin positive spiny neurons within the dorsomedial striatum; 22--28% of all opiate-peptide positive cells co-expressed NGFI-B mRNA. NGFI-B did not colocalize with striatal aspiny interneurons expressing choline acetyl transferase mRNA or those containing the calcium-binding protein parvalbumin. The pattern of NGFI-B mRNA expression within different striatal spiny projecting neurons suggests that this transcription factor may have a direct effect on the function of different striatal efferent pathways. PMID- 11382385 TI - Separate urinary bladder and external urethral sphincter neurons in the central nervous system of the rat: simultaneous labeling with two immunohistochemically distinguishable pseudorabies viruses. AB - This work examines the distribution, in the central nervous system, of virus labeled neurons from the rat urinary bladder and the external urethral sphincter simultaneously within the same tissue sections. Two immunohistochemically distinct pseudorabies virus strains were injected into male Sprague--Dawley rats (approximately 280 g). One virus was injected into the bladder and the other into the external urethral sphincter. After incubation intervals of 2, 2.5 and 3 days, sections from the spinal cord and brain were treated immunohistochemically to detect cells which were labeled separately by each virus or were labeled by both viruses. The major result of these experiments is that each strain of virus labeled a separate population of neurons and that some neurons were labeled by both strains. In the lumbosacral cord, 3 days post-infection, neurons labeled by virus from the external urethral sphincter were found in Onuf's nucleus, the dorsal gray commissure, and the superficial dorsal horn. Neurons labeled by virus from the urinary bladder were found in the L6--S1 and L1--L2 spinal cord segments within the dorsal gray commissure, the intermediolateral area and the superficial dorsal horn. Double-labeled interneurons were mainly located in the dorsal gray commissure although some were also found in the intermediolateral area and the superficial dorsal horn. In the medulla, external urethral sphincter neurons and bladder neurons and double-labeled neurons were found in the reticular region and the raphe. More rostrally, bladder neurons were located in the pontine micturition center and external urethral sphincter neurons were found in the locus coeruleus and subcoeruleus. A very small number of double-labeled neurons were found in the pontine micturition center and the locus coeruleus or subcoeruleus. PMID- 11382386 TI - Regional expression of Bcl-2 mRNA and mitochondrial cytochrome c release after experimental brain injury in the rat. AB - Regional levels of anti-apoptotic Bcl-2 mRNA and the cytosolic cytochrome c protein were measured after lateral fluid percussion (FP) brain injury in rats. Levels of Bcl-2 mRNA were significantly decreased in the injured left cortex (IC) and ipsilateral hippocampus (IH), but not in the contralateral right cortex (CC) and hippocampus (CH) after brain injury. Levels of Bcl-2 mRNA were significantly decreased as early as 2 h and stayed decreased as long as 48 h in the IC and IH after injury. Levels of the cytosolic cytochrome c protein were significantly increased in the IC and IH, but not in the CC and CH after brain injury. Levels of cytosolic cytochrome c were significantly increased in the IC at 30 min, 48 and 72 h, and in the IH at 2 h and as long as 72 h after injury. The increase of cytosolic cytochrome c suggests that the mitochondrial release of cytochrome is increased in the IC and IH after lateral FP brain injury. These data show that the reduction of anti-apoptotic Bcl-2 and increases of mitochondrial release of cytochrome c protein occur only in the IC and IH, regions which have been observed to undergo apoptosis and neuronal cell loss after lateral FP brain injury. Therefore, it is likely that the reduction of Bcl-2 and the increased cytochrome c protein in the cytosol contribute to the observed apoptosis and neuronal cell death in the IC and IH after lateral FP brain injury in rats. PMID- 11382387 TI - Brainstem activates paroxysmal discharge in human generalized epilepsy. AB - In nine patients with generalized epilepsy of convulsive seizures, the excitability change of the brainstem was evaluated over the course of the interictal paroxysmal discharge (poly spike-and-wave complex, poly SWC). The evaluation was carried out by a sequential analysis of brainstem auditory evoked potentials (BAEPs) before and during one sequence of poly SWC. The characteristics of BAEPs, i.e. far-field evoked potentials, allowed the evaluation of the excitability change in the brainstem, which was not influenced by the cortical activity. The excitability in the ventral brainstem, measured with the parameters of wave-III, showed a biphasic fluctuation (deceleration- acceleration) before the onset of poly SWC (minima at -0.7+/-0.4 s). On the other hand, the excitability in the dorsal brainstem, measured with the parameters of wave-V, showed no significant difference over the course of poly SWC. The results suggest that the biphasic excitability change in the ventral brainstem is conveyed to the cortex through the ascending activating system. The excitability acceleration preceded by deceleration in the ventral brainstem probably synchronizes the cortical activity profoundly enough to produce poly SWC through the activation of intralaminar thalamic neurons. PMID- 11382388 TI - Naloxone inhibits nicotine-induced receptor current and catecholamine secretion in bovine chromaffin cells. AB - Nicotine-induced catecholamine (CA) secretion and inward ionic currents were inhibited by the opioid antagonist naloxone in cultured bovine chromaffin cells. Naloxone inhibited nicotine-induced CA secretion, as detected by an on-line real time electrochemical technique, in a dose-dependent manner (IC(50)=29 microM). In voltage-clamped chromaffin cells, nicotine (10 microM) evoked an average peak inward current of -146 pA that was inhibited by low concentrations of naloxone (42% at 0.1 microM). The antagonist also inhibited total charge influx associated with nicotinic receptor activation (53% at 0.1 microM). This provides strong evidence that naloxone modulation of nicotine-induced CA secretion does not involve opioid receptors but results from the direct interaction with the nicotinic receptor itself, which might also be the case for other related opioid compounds. PMID- 11382389 TI - Co-involvement of mitochondria and endoplasmic reticulum in regulation of apoptosis: changes in cytochrome c, Bcl-2 and Bax in the hippocampus of aluminum treated rabbits. AB - Neurodegenerative diseases, including Alzheimer's disease, are characterized by a progressive and selective loss of neurons. Apoptosis under mitochondrial control has been implicated in this neuronal death process, involving the release of cytochrome c into the cytoplasm and initiation of the apoptosis cascade. However, a growing body of evidence suggests an active role for the endoplasmic reticulum in regulating apoptosis, either independent of mitochondrial, or in concert with mitochondrial-initiated pathways. Members of the Bcl-2 family of proteins have been shown to either inhibit apoptosis, as is the case with Bcl-2, or to promote it, in the case of Bax. Investigations in our laboratory have focused on neuronal injury resulting from the intracisternal administration of aluminum maltolate to New Zealand white rabbits, an animal system relevant to a study of human disease in that it reflects many of the histological and biochemical changes associated with Alzheimer's disease. Here we report that treatment of young adult rabbits with aluminum maltolate induces both cytochrome c translocation into brain cytosol, and caspase-3 activation. Furthermore, as assessed by Western blot analysis, these effects are accompanied by a decrease in Bcl-2 and an increase in Bax reactivity in the endoplasmic reticulum. PMID- 11382390 TI - Nitric oxide synthase inhibitors enhance mechanosensitive Ca(2+) influx in cultured dorsal root ganglion neurons. AB - Nitric oxide (NO) can have opposite effects on peripheral sensory neuron sensitivity depending on the concentration and source of NO, and the experimental setting. The aim of this study was to determine the role of endogenous NO production in the regulation of mechanosensitive Ca(2+) influx of dorsal root ganglion (DRG) neurons. Adult mouse DRG neurons were grown in primary culture for 2-5 days, loaded with Fura-2, and tested for mechanically mediated changes in [Ca(2+)](i) by fluorescent ratio imaging. In the presence of the NOS inhibitors L NAME, TRIM, or 7-NI, but not the inactive analogue D-NAME, peak [Ca(2+)](i) transients to mechanical stimulation were increased more than 2-fold. Neither La(3+) (25 microM), an inhibitor of voltage activated Ca(2+) channels, or tetrodotoxin (TTX, 1 microM), a selective inhibitor of voltage-gated Na(+) channels, had an effect on mechanically activated [Ca(2+)](i) transients under control conditions. However, in the presence of L-NAME, both La(3+) and TTX partially blocked the [Ca(2+)](i) response. Addition of Gd(3+), a blocker of mechanosensitive cation channels and L-type Ca(2+) channels, at a concentration (100 microM) that markedly inhibited the mechanical response under control conditions, only partially inhibited the response in the presence of L-NAME. The combination of either La(3+) or TTX with Gd(3+) caused near complete inhibition of mechanically stimulated [Ca(2+)](i) transients in the presence of L-NAME. We conclude that focal mechanical stimulation of DRG neurons causes Ca(2+) influx occurs primarily through mechanosensitive cation channels under control conditions. In the presence of NOS inhibitors, additional Ca(2+) influx occurs through voltage-sensitive Ca(2+) channels. These results suggest that endogenously produced NO in cultured DRG neurons decreases mechanosensitivity by inhibiting voltage-gated Na(+) and Ca(2+) channels. PMID- 11382391 TI - A non-radioactive assay for the cAMP-dependent protein kinase activity in rat brain homogenates and age-related changes in hippocampus and cortex. AB - Cyclic AMP-dependent protein kinase (PKA) activity was involved in a number of brain functions such as cognitive process or aging. The measurement of PKA activity is traditionally based on the use of [(32)P]ATP in phosphorylation of specific protein. Recently non-isotopic PKA assays have been developed, but none has been tested on brain homogenates. This work aimed to adapt a fluorimetric method of PKA activity into a novel assay never applied before in brain homogenate, and to characterize the enzyme activity and ratio in hippocampus and cortex from rats of different ages. Optimal conditions of homogenization and enzyme protection were determined. The method was sensitive and reproducible (intra-assay and interassay variation was 5.0% and 9.0%, respectively). In hippocampal cytosol, PKA activity was 27+/-8 and 80+/-9 nmol/min per mg protein in basal and cAMP-stimulated activity, respectively, and accounted for 80% of total cell PKA activity. The non-PKA activity, assessed by the use of the PKA specific inhibitor (PKI) accounted for 49.0% and 65.0% of endogenous levels in cytosol and membrane, respectively. cAMP-augmenting drugs effects were measured and increase of 53%, 273% and 118% over basal by 10 microM isoproterenol, 100 microM forskolin, 1 microM Sp-AMP, respectively, was observed. With respect to the changes in animal age, PKA activity increased from newborn to the mature rats but decreased in older rats. The PKA ratio was higher in cytosol than in particulate fraction, and was decreased in hippocampal sample from old rats (P<0.05). This last result was interpreted as related to the loss of cognitive capacities in old animals. PMID- 11382392 TI - Changes in presynaptic proteins, SNAP-25 and synaptophysin, in the hippocampal CA1 area in ischemic gerbils. AB - A general consensus exists that the presynaptic terminals in the hippocampal CA1 area are resistant to ischemic stress in spite of the loss of their target cells (CA1 pyramidal neurons). We have verified this by immunostaining and Western immunoblotting using the antibodies for presynaptic proteins, synaptosomal associated protein of 25 kDa (SNAP-25) and synaptophysin in gerbils after bilateral carotid artery ligature. In the immunohistochemical analysis, decreases in SNAP-25 and synaptophysin immunoreactivities in the strata radiatum and oriens, especially around the apical dendrite of CA1 neurons, and disappearance of SNAP-25 immunoreactivity in the alveus were observed on day 2 after ischemia. On days 7 and 14, SNAP-25-positive granular materials were expressed in the CA1 area, and intense synaptophysin immunoreactivity around surviving CA1 neurons was observed. Western immunoblot analysis revealed significant decreases of SNAP-25 and synaptophysin (about 60% of control levels) on day 2, and then increase of their proteins (130--140% of control levels) on day 14. These results indicate that presynaptic degeneration occurs in the hippocampal CA1 area after ischemia, and it precedes the delayed neuronal death of CA1 neurons. The presynaptic terminal damage may be responsible for some pathological changes in ischemic brains. PMID- 11382393 TI - Dopamine output in the nucleus accumbens shell is related to the acquisition and the retention of a motivated appetitive behavior in rats. AB - Chronic stress exposure consistently impairs the reactivity to aversive and pleasurable stimuli in rats; these behavioral modifications are associated with a decrease in dopamine output in the nucleus accumbens shell (NAcS). However, when rats that have already acquired an appetitive behavior are exposed to chronic stress, they develop an impaired reactivity to avoidable aversive stimuli while retaining the appetitive behavior. The dissociation between these two behavioral traits was used to study whether the decreased dopaminergic activity in the NAcS was connected to either of the two deficits. Dopamine output was studied through microdialysis as dopamine accumulation following re-uptake inhibition by cocaine. When rats that had previously acquired the appetitive behavior were exposed to chronic stress, they showed a dopaminergic transmission in the NAcS similar to that of controls and significantly higher than that of chronically stressed animals. Thus, dopamine output in the NAcS was consistently associated to the acquisition and maintenance of appetitive behavior, while the expression of a deficit in avoidance appeared to be independent of it. PMID- 11382394 TI - Secondary somatosensory cortex stimulation facilitates the antinociceptive effect of the NO synthase inhibitor through suppression of spinal nociceptive neurons in the rat. AB - Electrical stimulation of the secondary somatosensory cortex (S-II), which is clinically effective in some chronic pain patients, produces a weak antinociception by itself and also strongly facilitates the antinociceptive effect of the neuronal NO synthase inhibitor 7-nitro-indazole in laboratory animals (rats). The present study thus investigated the mechanisms by which S-II stimulation facilitates the 7-nitro-indazole-induced antinociception. S-II stimulation in combination with 7-nitro-indazole at a subeffective dose, 5 mg/kg, synergistically reduced the number of cells expressing c-Fos in response to intraplantar injection of formalin in the superficial regions (laminae I and II) of the L4 and L5 spinal dorsal horn in conscious rats, although each had no significant effect. A similar synergism produced by S-II stimulation and 7-nitro indazole was also confirmed in both the first and second phases in the formalin induced behavioral nociception test. The synergistic antinociception exerted by S II stimulation in combination with 7-nitro-indazole was resistant to systemic administration of the opioid antagonist naloxone or the alpha-adrenoceptor antagonist phentolamine. In contrast, intrathecally administered methysergide, a serotonin receptor antagonist, at 20 microg/rat, abolished the first-phase, but not the second-phase, antinociception following S-II stimulation in combination with 7-nitro-indazole. These findings suggest that S-II stimulation, in combination with inhibition of neuronal NO synthase, can suppress spinal nociceptive neurons, at least in part through the descending spinal serotonergic pathway, resulting in antinociception. PMID- 11382395 TI - Central representation of the sympathetic nervous system in the cerebral cortex. AB - The sympathetic-related regions of the cerebral cortex were identified in rats after pseudorabies virus injections were made in functionally different targets: adrenal gland, stellate ganglion which regulates the heart, or celiac ganglion which innervates the gastrointestinal tract. Extensive transneuronal labeling was found in limbic system areas: (1) extended amygdaloid complex, (2) lateral septum, and (3) infralimbic, insular, and ventromedial temporal cortical regions (viz., ectorhinal cortex=Brodmann's area 36, perirhinal cortex=area 35, lateral entorhinal=area 28, and ventral temporal association cortex=Te3 region). Deep temporal lobe structures were prominently labeled, including the amygdalopiriform and amygdalohippocampal transition areas, ventral hippocampus and ventral subiculum. The cortical circuits mediating emotional-autonomic changes (i.e., mind-body control) are discussed. PMID- 11382396 TI - Expression and regulation of cholecystokinin and cholecystokinin receptors in rat nodose and dorsal root ganglia. AB - Cholecystokinin (CCK) is an important satiety factor, acting via the vagus nerve to influence central feeding centers. CCK binding sites have been demonstrated in the vagal sensory nodose ganglion and within the nerve proper. Using in situ hybridization, expression of the CCK(A) and (B) receptors (Rs), as well as of CCK itself, was studied in the normal nodose ganglion (NG), and after vagotomy, starvation and high-fat diet. CCK(A)-R mRNA expression in dorsal root ganglia (DRGs) was also explored. In the NG, 33% of the neuron profiles (NPs) contained CCK(A)-R mRNA and in 9% we observed CCK(B)-R mRNA. CCK mRNA was not found in normal NGs. Peripheral vagotomy decreased the number of CCK(A)-R mRNA-expressing NPs, dramatically increased the number of CCK(B)-R mRNA, and induced CCK mRNA and preproCCK-like immunoreactivity in nodose NPs. No significant differences in the number of NPs labelled for either mRNA species were detected following 48 h food deprivation or in rats fed a high-fat content diet. In DRGs, 10% of the NPs expressed CCK(A)-R mRNA, a number that was not affected by either axotomy or inflammation. This cell population was distinct from neurons expressing calcitonin gene-related peptide mRNA. These results demonstrate that the CCK(A)-R is expressed by both viscero- and somatosensory primary sensory neurons, supporting a role for this receptor as a mediator both of CCK-induced satiety and in sensory processing at the spinal level. The stimulation of CCK and CCK(B)-R gene expression following vagotomy suggests a possible involvement in the response to injury for these molecules. PMID- 11382397 TI - Metabolic mapping of brain regions associated with behavioral extinction in preweanling rats. AB - Fluorodeoxyglucose autoradiography, quantitative image analysis, and a multivariate tool (partial least squares) were used to assess distributed patterns of brain activation in postnatal day 17 and day 12 rat pups engaged in extinction of instrumental behavior. Pups were trained in a straight alley runway on an alternating reward schedule, or on a pseudorandom reward schedule, injected with fluorodeoxyglucose, and then shifted to continuous nonreward (extinction). Another group at each age served as handled controls. Day 17 pups trained on the alternating schedule demonstrated faster extinction rates compared to those trained on the pseudorandom schedule, a phenomenon known as the partial reinforcement extinction effect. No differences were found between day 12 groups. Partial least-squares analysis revealed age-related increases in fluorodeoxyglucose uptake across all three training conditions in the cingulate and frontal cortices, amygdala, midline thalamic nuclei, cerebellum, and in several brainstem regions. Training-related increases common to both age groups were found in the orbital frontal cortex, limbic thalamus, gigantocellular reticular nucleus, the somatosensory system, and cerebellum. Age-dependent training effects were found in the interpositus and medial cerebellar nuclei wherein fluorodeoxyglucose uptake increased in the day 12 alternation and pseudorandom groups relative to controls. Day 12 pups trained on the alternating schedule demonstrated increased uptake in the anterior dorsal thalamus relative to pseudorandom and control pups. Hence, a large-scale neural system comprised by somatosensory, cerebellar, and brainstem regions govern extinction behavior in preweanling rats. Recruitment of limbic structures may allow the older pups to modify extinction behavior based on prior learning. PMID- 11382398 TI - Tyrosine hydroxylase-immunoreactive projections from the caudal ventrolateral medulla to the subfornical organ in the rat. AB - The subfornical organ (SFO) is known to be innervated by noradrenergic fibers. One possible origin of these fibers, which carry peripheral baroreceptor information to enhance the activity of SFO neurons, is the nucleus tractus solitarius (NTS). To investigate possible sites of origin of the catecholaminergic projections to the SFO, a retrograde tracing method was combined with immunohistochemistry in the rat. Stereotaxical injection of a retrograde tracer, wheat germ agglutinin-conjugated horseradish peroxidase- colloidal gold complex, into the SFO from the dorsal aspect revealed retrogradely labeled neurons in several catecholaminergic cell groups. A substantial number of retrogradely labeled neurons showing tyrosine hydroxylase (TH) immunoreactivity were found in the NTS and ventrolateral medulla (VLM) at levels caudal to the obex and in the locus coeruleus, while retrogradely labeled neurons without TH immunoreactivity were found in the VLM at levels rostral to the obex and in the nucleus prepositus hypoglossi. When the tracer was injected into the structures dorsal to the SFO, including the triangular septal nucleus, the frequency of retrogradely labeled neurons in the NTS and VLM at the caudal level was very low. These findings indicate the existence of catecholaminergic projections from the VLM (probably A1) to the SFO, in addition to the noradrenergic projections from the NTS previously reported. PMID- 11382399 TI - Kindling of the midbrain periaqueductal gray in rats. AB - We investigated the role of midbrain periaqueductal gray matter (PAG) in the manifestation of generalized seizures by administering electrical stimulation to this area in rats. Electrical stimulation of 60 Hz biphasic square pulses of a 1 s duration administered to the PAG-induced convulsive responses in the following order: (1) Type I, running (stimulus intensity; range 50--200 microA, mean 73.1 microA) without afterdischarge (AD), (2) Type IIa, running (stimulus intensity; range 50--300 microA, mean 111.8 microA) with AD at the PAG and the amygdala (AMY), and (3) Type IIb, generalized tonic--clonic seizures (GTCS) (stimulus intensity; range 50--250 microA, mean 182.1 microA) with AD at the PAG and AMY. Twenty daily PAG stimulations at the non-GTCS inducing threshold failed to produce kindling. However, 20 daily PAG stimulations at the GTCS threshold produced progressive AD spread involving the motor cortex, and progressive changes in the behavioral seizure pattern. These findings indicate that the PAG can be effectively kindled. However, PAG kindling has no apparent influence on subsequent AMY kindling. PMID- 11382400 TI - Organization of projections from the medial agranular cortex to the superior colliculus in the rat: a study using anterograde and retrograde tracing methods. AB - The organization of corticotectal projections from the medial agranular cortex (AGm), which has been considered to contain rat's frontal eye field, was examined using anterograde and retrograde tracing techniques. When biotinylated dextranamine (BDA) injections were made into the rostral part of the AGm, small numbers of BDA-labeled axons were found in the rostral two-thirds of the superior colliculus (SC) while some labeled axons were seen in the caudal one-third of the SC. These labeled axons were distributed mainly in the lateral part of the stratum griseum intermediale. On the other hand, after BDA injections into the caudal part of the AGm, moderate to dense plexuses of labeled axons were found in the rostral two-thirds of the SC while some labeled axons were seen in the caudal one-third of the SC. These labeled axons were distributed in the ventromedial and dorsolateral marginal zones of the stratum griseum intermediale as well as in the stratum griseum profundum. The corticotectal projections were largely uncrossed. After combined injections of BDA into the caudal part of the AGm on one side and cholera toxin B subunit (CTb) into the paramedian pontine reticular formation on the opposite side or into the interstitial nucleus of Cajal on the same side, the overlapping distributions of BDA-labeled axons and CTb-labeled neurons were found in the ventromedial marginal zone of the stratum griseum intermediale ipsilateral to the site of BDA injection. These results suggest that the caudal part of the AGm plays a more significant role in the oculomotor function than does the rostral part of the AGm. PMID- 11382401 TI - (S)-4C3HPG, a mixed group I mGlu receptor antagonist and a group II agonist, administered intrastriatally, counteracts parkinsonian-like muscle rigidity in rats. AB - The aim of the present study was to determine whether S-4-carboxy-3 hydroxyphenylglycine (S)-4C3HPG, a mixed group I glutamate metabotropic receptor antagonist and a group II agonist, attenuated parkinsonian-like muscle rigidity in rats. Muscle tone was examined using a combined mechano and electromyographic method, which measured simultaneously the muscle resistance (MMG) of the rat's hind foot to passive extension and flexion in the ankle joint and the electromyographic activity (EMG) of the antagonistic muscles of that joint: gastrocnemius and tibialis anterior. Muscle rigidity was induced by pretreatment with haloperidol (1 mg/kg i.p.). (S)-4C3HPG injected in doses of 5 and 15 microg/0.5 microl bilaterally, into the rostral region of the striatum, decreased both the haloperidol-induced muscle rigidity (MMG) and the enhanced electromyographic activity (EMG). The present results suggest that blockade of mGluR1 receptors and/or activation of mGluR2 ones, localized in the rostral part of the striatum, may be responsible for the anti-parkinsonian effect of (S) 4C3HPG. PMID- 11382402 TI - Axonal elongation through long acellular nerve segments depends on recruitment of phagocytic cells from the near-nerve environment. Electrophysiological and morphological studies in the cat. AB - The distal nerve stump plays a central role in the regeneration of peripheral nerve but the relative importance of cellular and humoral factors is not clear. We have studied this question by freezing the tibial nerve distal to a crush lesion in cat. The importance of constituents from the near-nerve environment was assessed by modification of the contact between the tibial nerve and the environment. Silicone cuffs, containing electrodes for electrophysiological assessment of nerve regeneration, were placed around the tibial nerve distal to the crush site. The interaction between long acellular frozen nerve segments (ANS) and the near-nerve environment was ascertained by breaching the silicone cuff to allow access of cellular or humoral components. Tibial nerves were crushed and frozen for 40 mm and enclosed in nerve cuffs with 0.45-microm holes or 2.0-mm holes to allow access of humoral factors or tissue ingrowth, respectively. In a second set of experiments, tibial nerves were crushed and either frozen for 20+20 mm, leaving a 10 mm segment with viable cells in the center (stepping-stone segment) or frozen for 50 mm. These nerves were enclosed in cuffs with 2.0 mm holes corresponding to the viable nerve segment. The regeneration was monitored electrophysiologically by implanted electrodes and after 2 months the nerves were investigated by light and electron microscopy. The results indicate that soluble substances in the near-nerve environment, such as nutrients, oxygen or tropic substances did not exert any independent beneficial effect on the outgrowing axons. However, phagocytic cells entering the acellular segment from the near-nerve environment were crucial for axonal outgrowth in long ANS. PMID- 11382403 TI - Firing patterns of pre-Botzinger and Botzinger neurons during hypocapnia in the adult rat. AB - Controversy exists about how a coordinated respiratory rhythm is generated in the brainstem. Some authors suggest that neurons in the pre-Botzinger complex are key to initiation of all types of breathing. While, on the other hand, it has been reported that some pre-Botzinger neurons fail to maintain a rhythmic discharge in phase with phrenic nerve discharge during mechanical hyperventilation. Extracellular recordings were made from respiratory units in the pre-Botzinger and Botzinger complexes of 13 anaesthetised, paralysed and vagotomised rats. Central respiratory activity was monitored from the C5 phrenic nerve. During mechanical hyperventilation, several changes were observed in the phrenic neurogram. Firstly, the frequency and amplitude of integrated phrenic nerve discharge were reduced and reversibly stopped. Secondly, the patterned discharges changed from an augmenting to a variety of non-augmenting patterns in 53 of 60 cases. In some cases (n=9) we observed that the pattern appeared to have two components, an early short duration discharge followed by a longer duration discharge. Respiratory units also started to show different firing patterns during mechanical hyperventilation. In general, they were divided into those units that fired tonically (n=28) and units that became silent (n=32), before phrenic nerve discharge ceased coincidently with complete apnoea. Of particular interest were those expiratory-inspiratory units in the pre-Botzinger complex (n=8) that narrowed their firing period towards late expiration and early inspiration during mechanical hyperventilation. Given their firing features, it is possible that these expiratory-inspiratory units may participate in generation of the early inspiratory component of phrenic nerve discharge. PMID- 11382405 TI - Opioid neurotransmission in the post-ictal analgesia: involvement of mu(1)-opioid receptor. AB - Pentylenetetrazol (PTZ), a non-competitive antagonist that blocks GABA-mediated Cl(-) flux, was used in the present work to induce seizures in animals. The aim of this work is to study the neurochemical basis of the antinociception induced by convulsions elicited by peripheral administration of PTZ (64 mg/kg). The analgesia was measured by the tail-flick test, in eight rats per group. Convulsions were followed by significative increase in the tail-flick latencies (TFL), for at least 120 min of the post-ictal period. Peripheral administration of naltrexone (5 mg/kg, 10 mg/kg and 20 mg/kg) caused a significant decrease in the TFL in seizing animals, as compared to controls. These data were corroborated with peripheral administration of naloxonazine (10 mg/kg and 20 mg/kg), a mu(1) opioid blocker, in the same doses used for non-specific antagonist. These results indicate that endogenous opioids may be involved in the post-ictal analgesia. The involvement of mu(1)-opioid receptor was also considered. PMID- 11382404 TI - Stimulation of astrocyte-enriched culture with C2 ceramide increases proenkephalin mRNA: involvement of cAMP-response element binding protein and mitogen activated protein kinases. AB - In rat astrocyte-enriched culture, C2 ceramide dose- and time-dependently increased proenkephalin (proENK) mRNA; the significant increase began at 6 h after 30 microM C2 ceramide treatment (about 13-fold) and at 12 h after treatment (about 21-fold). In addition, C2 ceramide also increased AP-1 proteins, such as Fra-1, c-Jun, JunB and JunD, and phosphorylation of CREB. The blocking of protein synthesis by cycloheximide (CHX) evokes a further increase of C2 ceramide-induced proENK mRNA and phospho-CREB level, while C2 ceramide-induced increases of AP-1 protein levels were reduced by CHX. The C2 ceramide-induced proENK mRNA expression was not changed significantly by the pretreatment with H89 (a PKA inhibitor), KN62 (a calcium/calmodulin-dependent protein kinase II inhibitor), and PD98059 (an ERK pathway inhibitor). However, calphostin C (a PKC inhibitor) and or SB203580 (a p38 inhibitor) partially but significantly reduced C2 ceramide induced proENK mRNA expression as well as phospho-CREB level. These results suggest that, in the rat astrocyte-enriched culture, C2 ceramide increases proENK mRNA expression via phosphorylation of CREB rather than the increases of AP-1 protein levels. Additionally, the activations of PKC and p38, but not PKA, calcium/calmodulin-dependent protein kinase II, and ERK, by C2 ceramide play important regulatory roles in C2 ceramide-induced proENK mRNA expression via activating the CREB. PMID- 11382406 TI - Effects of riluzole on N-methyl-D-aspartate-induced tyrosine phosphorylation in the rat hippocampus. AB - Since increased tyrosine phosphorylation has been observed in response to brain ischemia, we investigated whether riluzole (an inhibitor of glutamate neurotransmission with neuroprotective properties) affects tyrosine phosphorylation stimulated by N-methyl-D-aspartate (NMDA) in rat hippocampal slices. Riluzole produced an extremely potent concentration-related inhibition of NMDA (1 mM)-stimulated protein tyrosine phosphorylation (IC(50)=0.5+/-0.03 microM, mean+/-S.D.), but failed to affect that evoked by phorbol 12-myristate 13 acetate (PMA, an activator of protein kinase C, 0.1 and 1 microM). These results suggest that inhibition of tyrosine phosphorylation may contribute to the neuroprotective effects of riluzole against excitotoxic injury. PMID- 11382407 TI - Cholinergic- and stress-induced signaling activities in cells overexpressing wild type and mutant presenilin-1. AB - This study examined the effects of overexpression of presenilin-1 wild-type (PS1wt) or mutant L286V (PS1m) in human neuroblastoma SH-SY5Y cells on signal transduction systems. Oxotremorine-M-induced activation of AP-1 was 40--53% lower in PS1wt than control cells, and further impaired (63--76%) in PS1m cells. Heat shock (45 degrees C) activated Akt, increased heat shock factor-1 (HSF-1) DNA binding activity, and increased levels of heat shock protein 70, and these responses were not altered by overexpression of PS1wt or PS1m. H(2)O(2) also caused a time-dependent increase in HSF-1 DNA binding activity which was similar in all cell lines. Thus, overexpression of PS1wt reduced muscarinic receptor mediated activation of AP-1, and PS1m overexpression caused greater inhibition, but stress-induced activation of Akt and HSF-1 was unaffected by either PS1wt or PS1m. PMID- 11382408 TI - Distribution of various calcium channel alpha(1) subunits in murine DRG neurons and antinociceptive effect of omega-conotoxin SVIB in mice. AB - Immunohistological study revealed the differential localization of subtypes of voltage-dependent calcium channels in the dorsal root ganglion neurons. Intrathecal injection of omega-conotoxin SVIB, an analogue of omega-conotoxin GVIA, which acts on N-type voltage-dependent calcium channels, significantly shortened the licking time in the late phase of a formalin test. PMID- 11382409 TI - Towards micro electrode implants: in vitro guidance of rat spinal cord neurites through polyimide sieves by Schwann cells. AB - Our goal is to develop biohybrid neural microprobe implants with sieve electrodes for external stimulation of co-implanted neurons whose axons penetrate through the holes of electrodes and innervate host targets such as denervated muscle fibers. For evaluation of implants, potential scar formation was imitated in fibroblast-spinal cord co-cultures. In vitro neurite extension through flexible 10-microm thick polyimide sieves was inhibited by co-cultured fibroblasts. In contrast, the neurite penetration of sieves could be greatly stimulated by oriented exposure to Schwann cells. To our knowledge this is the first direct proof that Schwann cells display a guidance effect on spinal cord neurons in vitro. The results pave the way for novel biohybrid neuro-implants and provide means to circumvent the obstacle of inhibitory scar formation. PMID- 11382410 TI - Changes in defensive behaviors following olfactory bulbectomy in male and female rats. AB - The present study examined if olfactory bulbectomy (OBX) altered defensive behaviors on the elevated plus-maze and the open-field differently in male and female rats. Similar increases in defensive behaviors in male and female rats were observed in both tests following OBX. No significant correlations were detected between defensive behaviors and activity, supporting the hypothesis that some behavioral changes following OBX may be due to decreased defensive behaviors and not increased activity. PMID- 11382411 TI - Differential alterations in the distribution of voltage-gated calcium channels in aged rat cerebellum. AB - In the present study, we used immunohistochemistry and Western blot analysis to determine region-specific changes in the distribution of voltage-gated calcium channels (VGCCs) in aged rat cerebellum. Age-dependent changes in the staining intensity of the alpha(1C) and alpha(1D) subunits were prominent in the Purkinje cells, whereas there was no change in the expression of the alpha(1A) and alpha(1B) subunits. In the aged rat, in particular, immunoreactivity for the alpha(1C) subunits were increased in the dendrites as well as in the cell bodies of Purkinje cells. On the other hand, decreases in immunoreactivity for alpha(1A) and alpha(1D) subunits were found in the molecular or granular layers. However, only alpha(1D) subunit immunoreactivity was decreased in the aged cerebellum membrane by Western blot analysis that, while not addressing regional specificity, further confirmed an age-related decrease in alpha(1D) subunit. These age-related changes in alpha(1D) subunit expression might reflect a gradual loss of regulation for L-type Ca(2+) channels in the senescent period. The first demonstration of age-related alterations in VGCC expression may provide useful data for future investigations on aging and neurodegenerative diseases. PMID- 11382412 TI - Time dependent amelioration against ischemic brain damage by glial cell line derived neurotrophic factor after transient middle cerebral artery occlusion in rat. AB - Time dependent influence of glial cell line-derived neurotrophic factor (GDNF) was examined after 90 min of transient middle cerebral artery occlusion (MCAO) in rats. Treatment with GDNF significantly reduced the infarct volume stained with 2,3,5-triphenyltetrazolium chloride (TTC) when GDNF was topically applied at 0 and 1 h of reperfusion, but became insignificant at 3 h as compared to vehicle group. The protective effect of GDNF was closely related to the significant reduction of the number of terminal deoxynucleotidyl transferase-mediated dUTP biotin in situ nick end labeling (TUNEL) positive cells as well as immunofluorescently positive cells for active forms of caspases, especially active caspase-3 but not -9. Thus, the present study showed that topical application of GDNF significantly reduced infarct size in a time-dependent manner, while the therapeutic time window was shorter than other chemical compounds such as an NMDA receptor antagonist (MK-801) and a free radical scavenger (alpha-phenyl-tert-butyl-nitrone, PBN). The effect of GDNF was stronger in suppressing active caspase-3 than active caspase-9. PMID- 11382413 TI - Hypocretin (orexin) input to trigeminal and hypoglossal motoneurons in the cat: a double-labeling immunohistochemical study. AB - In trigeminal and hypoglossal motor nuclei of adult cats, hypocretin immunoreactive fiber varicosities were observed in apposition to retrogradely labeled motoneuron somata and dendrites. Among those lateral hypothalamus neurons that project to the hypoglossal nucleus some were determined to be hypocretin immunoreactive and were located amongst the single-labeled hypocretinergic neurons. These data suggest that hypocretin may play a role in the synaptic control of these motoneurons. PMID- 11382414 TI - Melatonin promotes sleep-like state in zebrafish. AB - The sleep-promoting effect of the pineal hormone melatonin in humans is known for decades. However, the mechanisms of this phenomenon remain obscure, mainly due to lack of a simple, genetically tractable, animal model. We now report that melatonin promotes sleep-like state in a diurnal lower vertebrate, zebrafish (Danio rerio), and this effect is mediated through activation of specific melatonin membrane receptors. Furthermore, our data show that the sleep-like state in zebrafish has fundamental similarities with sleep in mammals, including characteristic postures, elevated arousal threshold to sensory stimulation and a compensatory rest rebound following rest deprivation, and can be induced by conventional hypnotics, diazepam and sodium pentobarbital. Collectively, these data indicate that melatonin is evolutionary conserved sleep-promoting agent in diurnal species and suggest that zebrafish provide an efficient animal model for studying the molecular mechanisms of sleep regulation and for screening new types of hypnotic medications. PMID- 11382416 TI - A tract on track--an attack. PMID- 11382417 TI - Deprivation and violence in the community: a perspective from a UK Accident and Emergency Department. AB - Using routinely collected data from an Accident and Emergency department over a 12 month period, we demonstrated a strong correlation (Pearson Correlation Coefficient 0.90; 95% Confidence Intervals, 0.77-0.96) between material deprivation and attendance following violent assault. PMID- 11382418 TI - Facial fractures in a level I trauma centre: the importance of protective devices and alcohol abuse. AB - Urban trauma centres have recently noted a shift in the causative mechanism of facial fractures away from motor vehicle crashes (MVC) to blunt assaults (BA). This study was conducted to examine the incidence and aetiology of facial fractures at our institution as well as the relationship with alcohol and protective device use. Trauma registry records of all patients admitted to a level I trauma centre from 1 January 1988 to 1 January 1999 were reviewed. There were 13594 trauma admissions during the 11-year period. Facial fractures were sustained by 1429 patients (10.5%) and this group forms the subject of this study. MVC was the predominant aetiology (59.9%) followed by BA (18.8%). Facial fractures were found in 9.5% of restrained MVC patients compared to 15.4% of unrestrained patients (P<0.00l). Non-helmeted motorcyclists were four times more likely to sustain facial fractures (4.3% vs. 18.4%) than helmeted patients (P<0.00l). 39.6% of patients in the MVC group were legally intoxicated compared to 73.5% in the BA group (P<0.00l). 45.4% of unrestrained patients with facial fractures were intoxicated compared to 11.8% of restrained MVC patients with facial fractures (P<0.001). MVC continue to be the primary aetiology of facial fractures in our trauma population. Protective devices decrease the incidence of facial fractures. Lack of protective device use and the consumption of alcohol correlate with sustaining facial fractures. PMID- 11382419 TI - Popliteal artery trauma. A critical appraisal of an uncommon injury. AB - Although popliteal artery injuries are uncommon, the consequent lack of management protocols may contribute to an already high level of outcome morbidity. We critically reviewed all nine cases of popliteal artery trauma treated at our institution in the last 5 years. The main findings were that blunt trauma was predominant and most patients presented with severe signs of ischaemia. Long delays occurred between injury and treatment, mostly due to the tyranny of distance. Nevertheless, most patients were subjected to further delay whilst undergoing unnecessary formal angiography. Interposition vein graft using contralateral long saphenous vein was the predominant procedure for arterial injury. Most patients did not receive systemic anticoagulation or fasciotomy, and none received intra-operative thrombolysis. Our limb salvage rate was 7/9, although two of these had persistent neurological disability. We criticise our shortcomings in the light of the current literature. PMID- 11382420 TI - Complex proximal humeral fractures in adults--a systematic review of management. AB - We performed a systematic review of the literature to compare the clinical outcomes of the management of three and four part fractures of the proximal humerus by conservative regimes, internal and external fixations and arthroplasty. We identified 147 reports of comparative trials and case series between 1969 and 1999. We selected only studies dealing exclusively with three and four part fractures of the proximal humerus, with at least 15 patients, treated within 48 h of injury by one of the three modalities studied, in which at least one of the outcomes of interest (pain, range of motion, infection and restoration of anatomy) was described. Also, to be included, studies had to have a follow-up period of at least 6 months, in which a minimum of 85% of patients were followed-up. Twenty four reports met our eligibility criteria. Conservatively managed patients had more pain and a poorer range of motion than those managed by either fixation or arthroplasty. Better restoration of anatomy was delivered in the fixation group. There was no significant difference in the functional range of motion between the arthroplasty and fixation groups, but there were very few studies available to compare infection rates in these two management modalities. The results from the present systematic review suggest that the data from the published literature are inadequate for evidence-based decision making with regards to the treatment of complex proximal humeral fractures. PMID- 11382421 TI - A new approach to the management of pretibial lacerations. AB - This paper describes a technique for the management of pretibial lacerations by deep reinforced suturing through steristrips, which are applied parallel to the wound edges. This is carried out under local anaesthesia and followed by application of gentle localised compression dressing. Typical victims of pretibial lacerations are the elderly and patients on long-term systemic steroid therapy. This suturing technique, which was used on both flap and linear lacerations, obliterates the dead space in the wound and prevents tearing of the thin, fragile skin of these patients. The dressing technique used, has a great advantage over the toes to knee pressure dressing currently used for such lacerations, because it frees the foot of bandaging and allows the patients (especially the elderly with decreased mobility) to wear their normal footwear immediately post-operatively and to maintain their normal mobility. In the total sample of 147 patients treated by this method, the average healing time was 26 days for 112 patients with flap lacerations, and 16 days for the remaining 35 patients with linear lacerations. This is significantly shorter than that reported in the medical literature using both non-operative methods and simple suturing. Moreover, none of these patients required skin grafting or hospitalisation (except for social reasons). PMID- 11382422 TI - Fluid accumulation and the rapid spread of bacteria in the pathogenesis of external fixator pin track infection. AB - An animal model was used to study the pathogenesis of pin track infection. The roles of fluid accumulation around the pin/bone interface and mechanical loosening of the pin were specifically studied. In addition, the spread of bacteria in relation to the clinical appearance of the pin track was assessed. This study demonstrated that fluid accumulation around the interface is an important factor in the spread of infection from the superficial wound track to the medulla of the bone. Spread occurs despite the absence of mechanical pin loosening, and can occur before any clinical features of infection are apparent. PMID- 11382423 TI - Treatment of unstable distal radial fractures by intrafocal, intramedullary K wires. AB - Displaced distal radial fractures with extensive dorsal comminution and plastic cancellous deformation are unstable and frequently cause treatment problems since there is no single, reliable method of treatment, notably in osteoporotic bone. We present a method of holding unstable distal radial fractures with blunt ended K-wires via intrafocal and intramedullary insertion, so modifying the Kapandji technique. Wires were placed dorsally, radially and when necessary from the volar direction depending on fracture configuration. Over a 7-year period (1992-1999) we treated 102 patients with unstable distal radial fractures in this way. Of these, 80 were followed up for 6-42 weeks according to clinical need and scored radiologically and functionally using modified Lidstrom scoring system. Results showed that 92 and 95% of these patients achieved good or excellent results in these scoring systems, respectively. We present this as a useful and reliable method of treating these common fractures, particularly in osteoporotic bone. PMID- 11382424 TI - Cerclage wiring in displaced both-column fractures of the acetabulum. AB - We evaluated the effect of cerclage wiring in the open reduction and internal fixation of displaced both-column fractures of the acetabulum. This was a prospective clinical evaluation of such cases where the main surgical strategy was open reduction and internal fixation with cerclage wire and supplemental reconstruction plates. Data on 35 cases treated by open reduction (all via the triradiate approach)/internal fixations with cerclage wire and reconstruction plates were collected. The follow-up period was 40 months (18-69). Reduction with a fracture gap of less than 2 mm without articular stepping was achieved in all 35 cases. Postoperative complications developed in seven cases, including subcutaneous haematoma in two, wound infection in two and heterotopic ossification in three. All the complications had no adverse effect on the clinical outcome, and all the cases had good to excellent final results. Cerclage wiring is very useful and effective in the reduction and fixation of displaced both-column fractures of the acetabulum, and supplemental fixation with reconstruction plates and screws is necessary. PMID- 11382425 TI - Dall-Miles cable and plate fixation for the treatment of peri-prosthetic femoral fractures-analysis of results in 13 cases. AB - We present a retrospective review of 13 patients with periprosthetic femoral fractures treated with the Dall-Miles cable and plate fixation system between 1995 and 1999. Of these, 12 fractures were in relation to hip arthroplasty and one was proximal to the femoral component of a total knee replacement. Mean age at presentation was 77 years (range 66-87) with the male female ratio of 6:7. All patients were followed up until there was evidence of clinical and radiological union. The average follow-up period was 6.5 months (range 2.5-24). We achieved satisfactory results in ten patients with average time to union of 4.4 months. The results were unsatisfactory in three patients who required further revision procedures due to failure of fixation or non-union. Internal fixation of the fracture with the loose femoral component left in-situ led to failure of union in one patient. Varus mal-alignment of the femoral component to the shaft of more than 6 degrees was associated with unsatisfactory outcome in two patients. The Dall-Miles cable and plate system is a useful method of internal fixation for majority of periprosthetic femoral fractures. This method may not be suitable if the femoral component is loose or if it is in varus angulation of more than 6 degrees to the shaft of the femur. PMID- 11382426 TI - The fatigue strength of small diameter tibial nails. AB - This study was performed to evaluate the fatigue strength of commonly used small diameter tibial nails. Five types of small diameter tibial nails (7 mm aap Biorigid Nail, 8 mm aap Biorigid Nail, 8 mm Ace UTN, 8 mm RT Tibial Nail, 8 mm Synthes UTN) with a sample size of 30 implants (six samples of each nail type) were tested with a new modular testing system. One module of the testing system was removed, simulating a 55-mm distal tibial defect, to induce maximum loading on the distal portion of the implant and locking bolts. The average yield strengths were obtained from static, single cycle tests and revealed an average static ultimate load of 1072 N (aap7), 1820 N (aap8), 1812 N (Ace), 1942 N (R&T), and 1543 N (Syn). The fatigue limits were evaluated by cyclic, sinusoidal loading and revealed a fatigue strength of 750 N (aap7), 1092 N (aap8), 906 N (Ace), 971 N (R&T), and 771 N (Syn) to endure 500000 cycles. Our results showed that the solid tibial nails fractured in the testing device in the same manner and location as they do in the clinical series. This study showed that all the small diameter tibial nails tested are obviously able to transmit loads of more than average body weight. The fatigue strength of the implants is relatively high, which means that patients without bone defects could be mobilised with full weight bearing as tolerated by pain. However, according to the results of this study, early mobilisation with full weight bearing in a normal gait cycle can not be suggested in patients with segmental tibial fractures or fractures with defect zones. PMID- 11382427 TI - Removal of the retained fragment of broken solid nails by the intra-medullary route. AB - The development of solid nails has not eliminated the risk of occasional nail failure. It has been suggested that subsequent intra-medullary removal of solid nail fragment may be virtually impossible and thus major surgery is required for their revision. We report two cases of hardware failure of a solid intra medullary nail (Synthes solid femoral and tibial nail) and describe their successful intra-medullary removal using the Synthes extraction kit. PMID- 11382428 TI - Outcome of compartment syndrome following intramedullary nailing of tibial diaphyseal fractures. AB - Seventeen cases of compartment syndrome were treated in a group of 626 consecutive patients with tibial diaphyseal fractures. Clinical and radiological follow-up was performed at an average of 24 months (range 8-54 months). Functional outcome was assessed using Edward's classification. All patients who developed compartment syndrome had fracture stabilisation with a reamed intramedullary nail using skeletal traction. The average interval between the nailing procedure and fasciotomy was 11 h. Results were good in 10 cases, fair in four cases and poor in the remaining three cases. Patients who had decompression within 12 h had a good functional outcome. Patients with poor results were all treated at an interval greater than 24 h. PMID- 11382429 TI - Routine monitoring of compartment pressure in patients with tibial fractures: Beware of overtreatment! AB - PURPOSE: This prospective clinical study sought to evaluate the possible effects of routine compartment pressure monitoring in tibial fractures and to assess the previously published criteria. Pressures in the anterior compartment were measured in consecutive cases of tibial fractures. In 95 patients, the protocol was completed with a follow-up after 1 yr. An optimal threshold pressure with both good sensitivity and specificity could not be identified. Such monitoring is a useful tool in the diagnosis of compartment syndrome, mainly in symptomatic patients or those who are difficult to assess, but routine monitoring and the use of most popular threshold pressures, which have low specificity, can result in overtreatment. PMID- 11382430 TI - Dynamised cast management of Achilles tendon ruptures. AB - Non-operative treatment of closed ruptures of the Achilles tendon within 48 h of injury has been shown to produce results comparable with surgical treatment [1,2]. We report 49 patients treated using a dynamised cast, with a mean follow up of 42 months, in which there has been only one case of re-rupture. A total of 33 patients were reviewed clinically and measurements revealed a mean calf circumference deficit of 1 cm and a mean functional range of ankle motion deficit of 3 degrees and plantar-flexion strength of 81-90%, when compared with the contra-lateral uninjured limb. Early mobilisation of the injured Achilles tendon using a dynamised cast has produced a re-rupture rate and functional recovery that compare favourably with previously published results for patients treated operatively. Non-operative management, in this way, is well tolerated by patients and is of low cost. PMID- 11382431 TI - A case report using Kirschner wires to facilitate closed reduction of a fracture dislocation of the elbow. PMID- 11382432 TI - Dislocation of cervico-thoracic spine with no neurological deficit--a case report. PMID- 11382516 TI - Galen and the origins of artificial ventilation, the arteries and the pulse. PMID- 11382517 TI - The respiratory system during resuscitation: a review of the history, risk of infection during assisted ventilation, respiratory mechanics, and ventilation strategies for patients with an unprotected airway. AB - The fear of acquiring infectious diseases has resulted in reluctance among healthcare professionals and the lay public to perform mouth-to-mouth ventilation. However, the benefit of basic life support for a patient in cardiopulmonary or respiratory arrest greatly outweighs the risk for secondary infection in the rescuer or the patient. The distribution of ventilation volume between lungs and stomach in the unprotected airway depends on patient variables such as lower oesophageal sphincter pressure, airway resistance and respiratory system compliance, and the technique applied while performing basic or advanced airway support, such as head position, inflation flow rate and time, which determine upper airway pressure. The combination of these variables determines gas distribution between the lungs and the oesophagus and subsequently, the stomach. During bag-valve-mask ventilation of patients in respiratory or cardiac arrest with oxygen supplementation (> or = 40% oxygen), a tidal volume of 6-7 ml kg(-1) ( approximately 500 ml) given over 1-2 s until the chest rises is recommended. For bag-valve-mask ventilation with room-air, a tidal volume of 10 ml kg(-1) (700-1000 ml) in an adult given over 2 s until the chest rises clearly is recommended. During mouth-to-mouth ventilation, a breath over 2 s sufficient to make the chest rise clearly (a tidal volume of approximately 10 ml kg(-1) approximately 700-1000 ml in an adult) is recommended. PMID- 11382518 TI - The evolutionary process of Medical Emergency Team (MET) implementation: reduction in unanticipated ICU transfers. AB - OBJECTIVE: To determine whether the introduction of the Medical Emergency Team (MET) system designed to provide immediate help for seriously ill patients: (i) changed the pattern of ICU patient transfers from the wards; and (ii) improved hospital survival rates. METHODS: Prospective information on MET calls and unanticipated ICU transfers was collected for 3 years in a suburban metropolitan hospital. RESULTS: A 3-year review of MET showed the number of MET calls doubled in the second and third year and the team was activated for more than just the most extremely ill patients. Whilst the frequency of calls for cardiopulmonary arrest remained constant (n = 16), increased use of the MET resulted in the proportion of calls for cardiopulmonary arrest dropping from 30% in year 1 to 13% in year 3. A slight decrease in the percentage of in-hospital deaths (0.74% in year 1 to 0.65% in year 3) was also demonstrated. The incidence of cardiopulmonary arrest per hospital admission also decreased slightly (0.08 0.07%). Although the overall number of ICU transfers remained constant, more seriously ill patients were transferred to ICU via the MET system. This was accompanied by a significant fall in unanticipated ICU transfers. Whilst the reduction in hospital deaths was encouraging, this study could not demonstrate whether the slight improvement in hospital survival rate over the 3 years was due to the MET system. CONCLUSION: More information is needed to demonstrate that the MET system improves patient survival. The study also highlights the importance of taking proactive measures, which should include providing in-service education on the benefits of early identification and treatment of patients who are at risk of acute deterioration, raising awareness and changing attitudes in hospitals when introducing system such as the MET. PMID- 11382519 TI - Results from the first 12 months of a fire first-responder program in Australia. AB - STUDY OBJECTIVE: We aimed to reduce response times and time to defibrillation for out-of-hospital cardiac arrest patients through fire first-responders equipped with automatic external defibrillators (AEDs). The fire first-responders were added as an extra tier to the existing two-tired ambulance response. METHODS: This prospective controlled trial set in Melbourne, Australia, consisted of a control area (277 km2, population density 2343/km2-ambulance only dispatch) and a pilot area (171 km2, population density 2290/km2-ambulance and fire first responder dispatch). The main outcome measures were time to emergency medical service (EMS) arrival at scene for all cardiac arrest patients and time to defibrillation for cardiac arrest patients presenting in ventricular fibrillation (VF). The study participants were patients who suffered a cardiac arrest of presumed cardiac aetiology for which a priority 0 emergency response was activated. A total of 268 patients were located in the control area and 161 in the pilot (intervention) area. RESULTS: The mean response time to arrival at scene was reduced by 1.60 (95% CI 1.21, 1.99) min, P < 0.001. A large reduction in prolonged responses (> or = 10 min) to cardiac arrests was also observed in the pilot area (2%) compared with the control area (18%), chi = 23.19, P < 0.001. Mean time to defibrillation was reduced by 1.43 (95% CI 0.11, 2.98) min, P = 0.068. CONCLUSION: The results from this study suggest that fire officers can be successfully trained in the use of AEDs and can integrate well into a medical response role. The combined response of ambulance and fire personnel significantly reduced the response interval and reduced time to defibrillation. This suggests that in appropriate situations other agencies could be considered for involvement in co-ordinated first-responder programs. PMID- 11382520 TI - Emergency medical service transport-induced stress? An experimental approach with healthy volunteers. AB - This randomized controlled trial was designed to evaluate the effects of simulated emergency medical service (EMS) transport related stress on hemodynamic variables, and catecholamine plasma levels. A total of 32 healthy male volunteers were randomized to being carried by paramedics from a third-floor apartment through a staircase with subsequent high-speed EMS transport with lights and sirens (stress; n = 16); or sitting on a chair for 5 min, and lying on a stretcher for 15 min (control; n = 16). Blood samples and hemodynamic variables were taken in the apartment before transfer, at the ground floor, and at the end of EMS transport in the stress group, and at corresponding time points in the control group. The stress versus control group had both significantly (P < 0.05) higher mean +/- SEM epinephrine (71 +/- 7 versus 37 +/- 3 pg/ml), and norepinephrine (397 +/- 29 versus 299 +/- 28 pg/ml) plasma levels after transport through the staircase. After EMS transport, the stress versus control group had significantly higher epinephrine (48 +/-6 versus 32 +/- 2 pg/ml), but not norepinephrine (214 +/- 20 versus 264 +/- 31 pg/ml) plasma levels. Heart rate increased significantly from 72 +/- 2 to 84 +/- 3 bpm after staircase transport, but not during and after EMS transport. In conclusion, volunteers being carried by paramedics through a staircase had a significant discharge of both epinephrine and norepinephrine resulting in increased heart rate, but only elevated epinephrine plasma levels during EMS transport. Transport through a staircase may reflect more stress than emergency EMS transport. PMID- 11382521 TI - Work of breathing characteristics of seven portable ventilators. AB - Portable ventilators (PVs) are used for patient transport with increasingly frequency. Due to design differences it would not be unexpected to find differences among these ventilators in the imposed work of breathing (WOBI) during spontaneous respiratory efforts. The purpose of this investigation was to compare the WOBI characteristics during spontaneous breathing of seven PVs; Bird Avian, Bio-Med Crossvent 4, Pulmonetics LTV 1000, Hamilton Max, Dragerwerk Oxylog 2000, Impact Uni-Vent 750, and Impact Uni-Vent 754 using a model of spontaneous breathing. Differences between the PVs in regards to the measured parameters increased with increases in simulated breathing demand. WOBI, peak inspiratory pressure, and pressure-time product were consistently less with the LTV 1000 over the range of simulated breathing conditions. During pressure support ventilation these parameters were significantly less with the LTV 1000 compared with the Crossvent 4. Only the WOBI produced by the LTV was consistently lower than the physiologic work of breathing across the simulated spontaneous breathing conditions. Based on these results it is predicted PVs with flow triggering and positive end-expiratory pressure compensation will consistently offer the least WOBI. Clinicians should be aware of these characteristics when using PVs with spontaneous breathing patients. PMID- 11382522 TI - Ejection as a key word for the dispatch of a physician staffed helicopter: the Swiss experience. AB - We evaluated the type and severity of injuries and the possible influence of a helicopter staffed by a physician on the outcome of 71 consecutive occupants ejected from a four-wheel vehicle ejected occupants who were cared for by the Swiss Air Rescue Helicopter team from January 1994 to February 1999. The investigation and the data collection were planned prospectively. The following data were collected for each patient ejected from a four wheel vehicle: (1) demographic information; (2) type of injury; (3) vital signs on scene, in flight and at hospital; (4) hospital diagnosis; (5) injury severity score; (6) secondary transfer; (7) length of stay in hospital and on intensive care; and (8) outcome at hospital discharge. A control group included consecutive patients cared for by the same rescue team during the same period but who were not ejected out of their vehicle. Forty-four percent of the ejected patients had a GCS < or = 8, 21% were hypotensive and 22% had respiratory problems. Nine patients died at the scene. A total of 53% of the 62 ejected patients who were transported had an ISS > or = 16. The median ISS was 17. A total of 37% of the patients were intubated at the scene, needle chest decompression was performed in 5% and major analgesia was used in 27% of the cases. A total of 38% of the patients needed surgery in the first 4 h, 34% needed intensive care. No patient needed secondary transfer to the Trauma Centre if they were not brought there in the first instance. The outcome was poor in 27 cases (38%): 17 died and 10 needed transfer to specialised institutions. Non-ejected patients suffered mostly from head and neck injuries (50%) of which 9% were severe (head and neck AIS > or = 4, P < 0.05). Thoracic injuries were less frequent (35%) of which 13% were severe (thorax AIS > or = 4, P < 0,05). The median ISS was 9 for the non-ejected patients, P < 0.05. In conclusion, ejection from a four-wheel vehicle causes more severe injuries and requires a high number of advanced life support manoeuvres. Based on the mechanism of injury alone, patients ejected from four-wheel vehicles should automatically receive a response from the best available pre-hospital team. In our system, this means the dispatch of a physician staffed helicopter. PMID- 11382523 TI - Communication between members of the cardiac arrest team--a postal survey. AB - AIMS: Effective communication enhances team building and is perceived to improve the quality of team performance. A recent publication from the Resuscitation Council (UK) has highlighted this fact and recommended that cardiac arrest team members make contact daily. We wished to identify how often members of this team communicate prior to a cardiopulmonary arrest. METHOD: A questionnaire on cardiac arrest team composition, leadership, communication and debriefing was distributed nationally to Resuscitation Training Officers (RTOs) and their responses analysed. RESULTS: One hundred and thirty (55%) RTOs replied. Physicians and anaesthetists were the most prominent members of the team. The Medical Senior House Officer is usually nominated as the team leader. Eighty-seven centres (67%) have no communication between team members prior to attending a cardiopulmonary arrest. In 33%, communication occurs but is either informal or fortuitous. The RTOs felt that communication is important to enhance team dynamics and optimise task allocation. Only 7% achieve a formal debrief following a cardiac arrest. CONCLUSION: Communication between members of the cardiac arrest team before and after a cardiac arrest is poor. Training and development of these skills may improve performance and should be prioritised. Team leadership does not necessarily reflect experience or training. PMID- 11382524 TI - Emergency drug availability for the cardiac arrest team: a national audit. AB - Cardiac arrest teams are called upon to deal with many different acute medical emergencies, including cardiac arrest. However, the drugs that are supplied for them to perform their role differs vastly from hospital to hospital. We have confirmed this in an audit of adult cardiac arrest teams from all the acute hospitals within Wales. The rational use of defibrillation and drugs during cardiopulmonary resuscitation has been standardised according to International guidelines, and there is no reason why resuscitation kits could not also be standardised. Further considerations of drug availability is needed for acute management of other collapsed patients without cardiac arrest, most commonly vaso vagal syncope, anaphylaxis, hypoglycaemia, fits and respiratory arrest. Based upon recent recommendations from the project team of the Resuscitation Council (UK), augmented by other commonly needed drugs for first aid in the collapsed patient, we propose a reduced and simplified emergency drug list, employing a two box system covering immediate and most secondary requirements. A standardised format of drug kits for use by adult cardiac arrest teams could speed effective delivery of emergency care. PMID- 11382525 TI - Serum levels of the brain-derived proteins S-100 and NSE predict long-term outcome after cardiac arrest. AB - BACKGROUND AND PURPOSE: patients with cardiac arrest have a high mortality and the long-term outcome is doubtful. The prognosis is mainly dependent on clinical parameters. S-100 and neurone specific enolase (NSE) are established biochemical markers of central nervous system (CNS) injury. The purpose of this study was to validate the use of serum determinations of S-100 and NSE with neurological investigations in regard to brain damage and long-term outcome after cardiac arrest. METHODS: neurological examinations were performed on 66 patients after cardiac arrest. Serum levels of S-100 and NSE were determined during the first 3 days of post arrest, using commercial luminescent immunoassays (LIAs). The main outcome variable was the Glasgow Outcome Scale (GOS), while secondary variables were the activity of daily living (ADL) index and mini mental state examination (MMSE). Outcome was determined at 1 year. RESULTS: the serum levels of S-100 and NSE were increased during the first 3 days after the arrest and were related to coma depth, time of anoxia and abnormal brain stem reflexes. High levels predicted a poor outcome, according to the GOS (death, vegetative state and severe disability). The prognostic value of the brain damage markers was comparable with that of traditional clinical parameters. None of the secondary outcome variables (ADL and MMSE) was strongly associated with S-100 or NSE. DISCUSSION: the serum levels of S-100 and NSE increased after cardiac arrest due to the anoxic brain damage. The determination of S-100 and NSE can be used as an adjunct to predict long-term outcome after cardiac arrest. PMID- 11382526 TI - Early cellular brain damage and systemic inflammatory response after cardiopulmonary resuscitation or isolated severe head trauma: a comparative pilot study on common pathomechanisms. AB - Severe neurological deficits are common characteristics of patients surviving cardiopulmonary resuscitation (CPR) or isolated severe head trauma (SHT). For comparative evaluation of underlying pathomechanisms, 22 patients with out-of hospital cardiac arrest and successful CPR as well as 10 patients with SHT were included in our prospective study. Circulating S-100B was determined as an indicator of cellular brain damage. Interleukin-8 (IL-8), soluble E-selectin (sE selectin) and polymorphonuclear (PMN-) elastase were measured as markers of systemic inflammation following whole body ischaemia and reperfusion injury. Venous blood samples were drawn on scene (median time 11.0 min after starting basic life support) and in the intensive care unit (median time 12.5 h thereafter) in CPR patients and at admission to hospital (median time 43.8 min after trauma) and approx. 12 h later in SHT patients. Biochemical parameters in these samples were compared with specimens taken from 20 healthy volunteers. Initial median S-100B levels of the CPR and SHT patients were both significantly increased compared with the controls. Twelve hours later, significant falls in S 100B revealed no differences between the two patient groups, but did not reach control values. Median IL-8 and sE-selectin levels entry to the study were elevated in both patient groups compared with controls and showed further rises within the following 12 h. Finally, increased initial median levels of PMN elastase revealed significant differences between the patient groups and between patients and controls. Twelve hours later, median PMN-elastase values were equally elevated in the CPR and SHT subjects. Our preliminary data suggest similar pathomechanisms occurring after both CPR and SHT. Both clinical entities seem to be associated with early transient cellular brain damage as shown by prolonged rapidly increasing and subsequent fall in S-100B serum levels. In contrast, the prolonged elevation of circulating IL-8, sE-selectin and PMN elastase may indicate a very similar systemic inflammatory response by endothelial cells and neutrophils initiated by ischaemia and reperfusion injury in both conditions. Further studies should be carried out to determine the cause and the prognostic value of these biochemical parameters in relation to long-term neurological outcome. PMID- 11382527 TI - Mass cardiopulmonary resuscitation 99--survey results of a multi-organisational effort in public education in cardiopulmonary resuscitation. AB - Mass cardiopulmonary resuscitation (CPR) 99 in Singapore was a large-scale multi organisational effort to increase awareness and impart basic cardiac life support skills to the lay public. Mass CPR demonstrations followed by small group manikin practice with instructor guidance was conducted simultaneously in three centres, four times a day. The exercise enlisted 15 community organisations and received the support of 19 other organisations. Three hundred and ninety-eight manikins and 500 instructors ('I's) were mobilised to teach an audience of 6000 participants ('P's). Two surveys, for 'I's and 'P's were conducted with respondent rates of 65.8% and 50%, respectively. 73.6% of the P-respondents ('P R's) indicated that they attended the event to increase their knowledge. 66.9% were willing to attend a more comprehensive CPR course. Concerns and perceptions in performing bystander CPR were assessed. PMID- 11382528 TI - Long-term survival after in-hospital CPR. PMID- 11382529 TI - Out-of-hospital cardiac arrest in a child without overt cardiac disease: emergency department management. AB - This case report describes the successful resuscitation of a 7-year-old girl who had no previous history of cardiac disease other than one episode of syncope. She developed ventricular fibrillation for 10 min. External chest compressions, early defibrillation and orotracheal intubation were used with a successful outcome. PMID- 11382530 TI - Survival after cardiac arrest with a pH 6.6. PMID- 11382532 TI - Development and distribution of FMRFamide-like immunoreactivity in the toad (Bufo bufo) brain. AB - By using immunohistochemistry, we studied the development and distribution of the FMRFamide-like immunoreactive (ir) neuronal system in the toad brain during the ontogeny. In addition to this, experimental evidence was provided to show that the rostral forebrain-located FMRFamide neurons originate in the olfactory placode and then migrate into the brain along the olfactory pathway. During early development, within the brain, FMRFamide-ir perikarya first appeared in the periventricular hypothalamus. Later in development, FMRFamide-ir cells were visualized in the rostralmost forebrain simultaneously with similar ir cells in the developing olfactory mucosa. Selective ablation of the olfactory placode(s), prior to the appearance of the first FMRFamide-ir cells in the brain, resulted in the total absence of ir cells in the telencephalon (medial septum and mediobasal telencephalon) of the operated sides(s). The preoptic-suprachiasmatic infundibular hypothalamus-located FMRFamide-ir neurons were not affected by olfactory placodectomy, arguing that they do not originate in the placode. This result points to the placode as the sole source of such neurons in the rostral forebrain. PMID- 11382533 TI - Alteration of Bcl-2 expression in the nigrostriatal system after kainate injection with or without melatonin co-treatment. AB - In order to understand further the role of the anti-apoptotic Bcl-2 proto oncogene protein in excitotoxin-induced brain injury and possible interaction between Bcl-2 and the antioxidant melatonin, the expression of Bcl-2 in various brain parts was studied after intrastriatal injection of kainate (KA, 2.5 nmol) with or without co-treatment of melatonin (10 mg/kg, intraperitoneally (i.p.)). Three days after unilateral injection of KA to the striatum in the rat, a dramatic direct cytotoxic effect was observed, as indicated an expression of Bcl 2 immunoreactivity in TUNEL- and OX-42-positive cells in the KA-injected striatum and traumatized cortical region. A less severe detrimental effect was also observed in the ipsilateral substantia nigra and peritraumatic cortex, as reflected by an upregulation of Bcl-2-immunostained neurons. Surprisingly, a reduction in Bcl-2-immunoreactive neurons that was accompanied by a less severe loss of tyrosine hydroxylase-immunoreactive neurons in the nigrostriatal pathway was observed after co-treatment with melatonin. Western blot analysis confirmed that Bcl-2 expression is elevated in striatum and cortex on the lesioned side, and that its expression was attenuated substantially after systemic administration of melatonin. The results showing an upregulation of Bcl-2 in nigral neurons and reactive microglia after KA lesion are consistent with the view that Bcl-2 is protective in function in the central nervous system. PMID- 11382534 TI - Distribution of hypocretin-(orexin) immunoreactivity in the central nervous system of Syrian hamsters (Mesocricetus auratus). AB - The hypocretins are peptides synthesized in neurons of the hypothalamus. Recent studies have suggested a role for these peptides in the regulation of sleep, feeding, and endocrine regulation. The distribution of hypocretin-immunoreactive cell bodies and fibers has been extensively described in rats, but not in other species. This study was designed to examine the distribution of hypocretin immunoreactivity in Syrian hamsters, as important differences in neuropeptide distribution between rats and hamsters have previously been demonstrated. Immunoreactive cell bodies were found primarily in the lateral hypothalamic area and the perifornical area, although a few hypocretin-positive cells were also located in the dorsomedial hypothalamus and the retrochiasmatic area. Fibers were distributed throughout the brain in a pattern similar to that seen in rats. The densest projections were found in the paraventricular nucleus of the thalamus, locus coeruleus, dorsal raphe, and lateroanterior hypothalamus. The innervation of the anterior hypothalamus may be of particular interest as similar cluster of immunoreactivity does not appear to be present in rats. Moderate levels of immunoreactivity could be seen throughout the hypothalamus, the lateral septum, bed nucleus of the stria terminalis, A5 noradrenergic area, and the midline thalamic nuclei. Hypocretin-immunoreactive fibers are present in all lamina of the spinal cord, with the greatest axon densities in lamina 1 and 10. The widespread distribution of hypocretin suggests its involvement in a wide variety of physiological and behavioral processes. Our results in hamsters indicate that the organization of the hypocretin system is strongly conserved across species, suggesting an important role for the peptide and its projections. PMID- 11382535 TI - Parvalbumin-containing interneurons of the human cerebral cortex express nicotinic acetylcholine receptor proteins. AB - Cholinergic fibers from the basal forebrain are known to contact cholinoceptive cortical pyramidal neurons. Recent electrophysiological studies have revealed that nicotinic acetylcholine receptors are also present in human cerebrocortical interneurons. A direct visualization of nicotinic receptor subunits in cortical interneurons has, however, not yet been performed. We have applied double immunofluorescence using antibodies against parvalbumin --a marker for the Chandelier and basket cell subpopulation of interneurons--and to the alpha4 and alpha7 subunit proteins of the nicotinic acetylcholine receptor. The vast majority of the parvalbuminergic interneurons was immunoreactive for the alpha4 and the alpha7 nicotinic acetylcholine receptor. Provided these receptors would be functional--as suggested by recent electrophysiological findings--the connectivity pattern of cholinergic afferents appears much more complex than thought before. Not only direct cholinergic impact on cortical projection neurons but also the indirect modulation of these by cholinergic corticopetal fibers contacting intrinsic cortical cells would be possible. PMID- 11382537 TI - SPECT mapping of cerebral activity changes induced by repetitive transcranial magnetic stimulation in depressed patients. A pilot study. AB - Repetitive transcranial magnetic stimulation (rTMS) is being investigated as an alternative treatment for depression. However, little is known about the clinical role and the neurophysiological mechanisms of the action of rTMS in these patients. In this study, 99mTc-HMPAO single photon emission computed tomography (SPECT) was used to map the effects of left dorsolateral prefrontal rTMS on prefrontal activity in seven patients who met DSM-IV criteria for major depression resistant to pharmacological treatment. rTMS consisted of 30 trains of 2-s duration stimuli (20 Hz, 90% of motor threshold), separated by 30-s pauses. Each patient underwent three SPECTs: at baseline; during the first rTMS; and 1 week after 10 daily sessions of rTMS. Regional cerebral blood flow (rCBF) of each cerebral region was normalized to the rCBF value in the cerebellum and relative changes in normalized rCBF were addressed using a region-of-interest analysis. The Hamilton Depression Rating Scale (HDRS) was used for clinical evaluation before and after rTMS. A significant rCBF increase after the 10 sessions of rTMS was found in the left prefrontal region (MANOVA F=5.29, d.f.=2,10, P=0.027), but no significant rCBF changes were found during the first rTMS session. The remaining cerebral regions showed no significant rCBF changes at any time. Only two patients showed a clinical improvement after rTMS, with 50% reduction of the initial HDRS score. The study was repeated under placebo conditions (identical design but addressing coil discharges to the air) in these two patients, who failed to show any rCBF increase during sham-rTMS. No relationship was found between the percentage of left prefrontal rCBF change and the clinical findings. In conclusion, rTMS of the left prefrontal cortex induces a significant rCBF increase in this region, despite the limited clinical effect in our sample of depressed patients. Cerebral perfusion SPECT is a useful tool to map cerebral activity changes induced by rTMS. PMID- 11382538 TI - Diffusion tensor imaging (DTI) and proton magnetic resonance spectroscopy (1H MRS) in schizophrenic subjects and normal controls. AB - Several proton magnetic resonance spectroscopy (1H MRS) studies in schizophrenia have found reduced N-acetyl aspartate (NAA) concentrations in pre-frontal and temporal regions of the brain. Reductions in NAA may reflect abnormalities of neuronal structure (e.g. reduced neuronal density or viability) or abnormalities of neuronal function. Diffusion tensor imaging (DTI) measures diffusion anisotropy, an indicator of the structural integrity of a neuronal tract. Both techniques were used to examine the anatomical basis of pre-frontal dysfunction in schizophrenia. Ten patients with DSM-IV schizophrenia were compared with 10 healthy controls. 1H MRS and DTI were performed on a clinical MR system and analysed with a region of interest approach. NAA concentrations and diffusion anisotropy were measured in the same pre-frontal white matter region. Diffusion anisotropy was also measured in a control region (occipital white matter). 1H MRS revealed non-significant but consistently reduced NAA concentrations (by 10-15%) in the pre-frontal white matter in schizophrenic subjects. Diffusion anisotropy measures revealed no such differences between schizophrenics and controls. It is concluded that the abnormalities of 'connectivity' reported in brain-imaging studies of schizophrenia may not be attributable to structural abnormalities of white matter and that reduced NAA in the pre-frontal white matter may reflect abnormal function of structurally intact neurons. PMID- 11382539 TI - Phospholipid abnormalities in postmortem schizophrenic brains detected by 31P nuclear magnetic resonance spectroscopy: a preliminary study. AB - It has been hypothesized that schizophrenia arises from cell membrane abnormalities due to changes in phospholipid (PL) composition and metabolism. We have used high resolution, in vitro 31P nuclear magnetic resonance (NMR) to characterize the PLs in left frontal cortex (gray matter) of postmortem brain from four schizophrenics and five controls. High resolution 31P NMR spectra were obtained in an organic-solvent system to resolve PL classes (headgroups) and in a sodium-cholate, aqueous dispersion system to resolve phosphatidylcholine (PC) molecular species. Multivariate analysis which included the major PC molecular species and phosphatidylinositol (PI) showed a significant difference between schizophrenics and controls. Analysis of specific interactions showed that the PI was significantly higher in the schizophrenic group than in the control group. There were no differences between the two groups for other individual PL classes, or for individual PL subclasses determined by the linkage type at the sn-1 position on glycerol. There was a trend for total PL content to be higher in schizophrenics than in controls. There was no evidence for elevated lysophosphatidylcholine or lysophosphatidylethanolamine in schizophrenia. The intensity of the PC peak representing molecular species with one saturated and one unsaturated (one or two double bonds) acyl chain was higher for the schizophrenic group than for the control group. Although these results are not in complete agreement with previous studies, they support the idea that PL abnormalities occur in the brain in schizophrenia and that fatty acid metabolism may be abnormal. PMID- 11382540 TI - 31P Nuclear magnetic resonance spectroscopy findings in bipolar illness: a meta analysis. AB - Published literature comparing 31P MR brain spectra of bipolar patients to healthy controls was evaluated, focusing on phosphomonoester (PME)/phosphodiester (PDE) resonance areas because these metabolites are related to membrane phospholipids and membrane defects in bipolar disorder have been suggested. Studies comparing PME and/or PDE values of bipolar subjects to values observed in healthy controls were reviewed. Data from the studies meeting our inclusion criteria (8 reports involving 139 bipolar and 189 comparison subjects) were grouped according to the mood state of the subjects. Meta-analyses of data were performed to compare PME and PDE levels of euthymic bipolar patients to healthy controls, as well as comparing PME levels during euthymia in bipolar subjects to values observed during manic and depressed states. The PME values of euthymic bipolar patients were found to be significantly lower than PME values of healthy controls. Depressed bipolar patients had significantly higher PME values in comparison to euthymic bipolar patients. No significant difference could be detected between the PDE values of bipolars and controls. This meta-analysis found support for trait- and possibly state-dependent abnormalities of membrane phospholipid metabolism, which may reflect a dysregulation in brain-signal transduction systems of relevance in bipolar illness. PMID- 11382541 TI - Reliability of tissue volumes and their spatial distribution for segmented magnetic resonance images. AB - Before using MRI tissue segmentation in clinical studies as a dependent variable or as a means to correct functional data for differential tissue contribution, we must first establish the volume reliability and spatial distribution reproducibility of the segmentation method. Although several reports of volume reliability can be found in the literature, there are no articles assessing the reproducibility of the spatial distribution of tissue. In this report, we examine the validity, volume reliability, and spatial distribution reproducibility for our K-means cluster segmentation. Validation was examined by classifying gray matter, white matter, and CSF on images constructed using an MRI simulator and digital brain phantom, with percentage volume differences of less than 5% and spatial distribution overlaps greater than 0.94 (1.0 is perfect). We also segmented repeat scan MRIs from 10 healthy subjects, with intraclass correlation coefficients greater than 0.92 for cortical gray matter, white matter, sulcal CSF, and ventricular CSF. The original scans were also coregistered to the repeat scan of the same subject, and the spatial overlap for each tissue was then computed. Our overlaps ranged from 0.75 to 0.86 for these tissues. Our results support the use of K-means cluster segmentation, and the use of segmented structural MRIs to guide the analysis of functional and other images. PMID- 11382542 TI - Protective role of L-carnitine on liver and heart lipid peroxidation in atherosclerotic rats. AB - Lipid peroxides are considered to be the initiation factor for atherosclerosis. Present study depicts that L-carnitine treatment (300 mg/kg body weight/day) for 7 and 14 days caused significant reduction in the tissue lipid peroxidations. It also shows marked improvement in the antioxidant status. By this way carnitine maintain the normal function of the cells. PMID- 11382543 TI - Dietary n-3 fatty acids alter the contractile response to thromboxane A(2) agonists of porcine coronary arteries. AB - Dietary supplementation with marine fish oils rich in n-3 fatty acids reduces circulating thromboxane A(2) (TxA(2)). However, the effects on thomboxane A(2) receptor mediated vascular reactivity are uncertain. The aim of this study was to test the hypothesis that dietary modification of TxA(2) levels alters vascular responsiveness to TxA(2) analogues. Juvenile female white pigs were fed a diet enriched in either 5% (w/w) fish oil or beef tallow for 6 weeks. Serum and myocardial tissue levels of eicosapentaenoic and docosahexaenoic acid reached a plateau during this period. Vascular responses were measured in isolated coronary arterial rings with intact endothelium by isometric tension measurement. Arteries from pigs fed fish oil produced a greater maximum vasoconstrictor tension to the TxA(2) analogue U46619 than did rings from pigs fed beef tallow (120 +/- 6% compared to 92 +/- 8%, values represented as a percentage relative to the maximum vasoconstrictor effect obtained to KCl, regression analysis, analysis of variance, P or = 0.95). The mean IOP reduction from baseline in the brimonidine group (-2.8 +/- 2.8 mmHg) was not statistically different (P = 0.55) compared with the mean IOP reduction in the apraclonidine group (-3.6 +/- 3.3 mmHg). There were no statistically significant changes in mean heart rate or blood pressure in either group except for a slight reduction in diastolic blood pressure at 1 hour (P = 0.005) in the brimonidine group (-5.2 +/- 7.4 mmHg) compared with the apraclonidine group (-0.2 +/- 6.4 mmHg). There were no clinically significant side effects noted in either group. CONCLUSIONS: A single preoperative drop of brimonidine 0.2% is as effective as apraclonidine 0.5% in preventing IOP elevation immediately after anterior segment laser surgery. PMID- 11382626 TI - Intraoperative mitomycin-C without conjunctival and Tenon's capsule touch in primary trabeculectomy. AB - OBJECTIVE: To verify whether intraoperative mitomycin C (MMC) without conjunctival and Tenon's touch is effective in inhibiting the development of thin, avascular blebs in eyes undergoing primary trabeculectomy. DESIGN: Noncomparative, interventional study. PARTICIPANTS: Fifteen eyes of 15 consecutive patients undergoing primary trabeculectomy. INTERVENTION: All eyes underwent trabeculectomy with intraoperative MMC (0.25 mg/ml for 3 minutes) without either conjunctival or Tenon's touch. Patients were examined 1 month, 3 months, 6 months, and 12 months after surgery. Intraocular pressure (IOP) and number of medications were evaluated at each examination. The appearance of the bleb was classified at the last examination into one of three groups: flat and vascularized; elevated but not avascular; or elevated, thin, and avascular. MAIN OUTCOME MEASURES: Intraocular pressure, number of antiglaucoma medications, and appearance of the bleb. RESULTS: Preoperative mean IOP was 30.57 +/- 10.92 mmHG: Statistically significant IOP reductions were observed 1 month, 3 months, 6 months, and 1 year after surgery (P < 0.01). Twelve months after surgery, the mean IOP was 14.92 +/- 6.53 mmHG: Five eyes (33.3%) showed an IOP less than 15 mmHg without antiglaucoma medication at the 12-month examination. The bleb was considered elevated, thin, and avascular in 12 of 15 eyes (80%) at the end of follow-up. CONCLUSIONS: Intraoperative MMC at 0.25 mg/ml for 3 minutes without either conjunctival or Tenon's touch was not effective in eliminating the development of thin, avascular blebs in eyes undergoing primary trabeculectomy. PMID- 11382627 TI - Ocular hemodynamics in pseudoexfoliation syndrome and pseudoexfoliation glaucoma. AB - OBJECTIVE: To evaluate orbital blood flow velocities by using color Doppler imaging in patients with pseudoexfoliation syndrome and pseudoexfoliation glaucoma. DESIGN: Prospective, comparative case series. PARTICIPANTS: Twenty eight patients with pseudoexfoliation syndrome (n = 14) or pseudoexfoliation glaucoma (n = 14) and 14 healthy control participants were included. INTERVENTION: Color Doppler imaging was used with a 7.5-MHz probe. Evaluation of the ophthalmic, central retinal, short posterior nasal, and temporal ciliary arteries was performed, and peak systolic and end diastolic flow velocities were measured. Resistive indices were calculated. RESULTS: When compared with the control participants, patients with pseudoexfoliation syndrome showed statistically significant decreases in the mean peak systolic velocity of the central retinal artery (11.21 +/- 2.19 cm/second; P < 0.05), and end diastolic velocities of the central retinal artery (3.00 +/- 1.03 cm/second; P < 0.005), and short posterior temporal ciliary arteries (3.50 +/- 1.74 cm/second; P < 0.005), whereas mean resistive indices of the ophthalmic artery (0.75 +/- 0.06 cm/second; P < 0.005) and central retinal artery were found to have increased (0.70 +/- 0.05 cm/second; P < 0.01). Patients with pseudoexfoliation glaucoma, when compared with the control participants, showed statistically significant decreases in the mean peak systolic and end diastolic velocities and increased mean resistive indices in all vessels measured (P < 0.05). Compared with the patients with pseudoexfoliation syndrome, patients with pseudoexfoliation glaucoma showed statistically significant decreases in the mean peak systolic velocities of the ophthalmic artery (30.07 +/- 4.00 cm/second; P < 0.05) and short posterior nasal ciliary arteries (2.35 +/- 0.09 cm/second; P < 0.05), and in the mean end diastolic velocities of the ophthalmic artery (6.28 +/- 2.12 cm/second; P < 0.05), and short posterior nasal ciliary arteries (2.35 +/- 0.09 cm/second; P < 0.05). The differences in the mean resistive indices were not statistically significant between the patients with pseudoexfoliation syndrome and the ones with pseudoexfoliation glaucoma. CONCLUSIONS: The findings suggest that hemodynamic parameters in the retrobulbar vessels were altered in patients with pseudoexfoliation syndrome and pseudoexfoliation glaucoma; however, these alterations were more prominent in the latter group. PMID- 11382628 TI - Pars plana tube insertion of glaucoma drainage implants and penetrating keratoplasty in patients with coexisting glaucoma and corneal disease. AB - PURPOSE: To determine the efficacy and associated complications of glaucoma drainage implant (GDI) surgery with pars plana tube insertion and penetrating keratoplasty (PK) in eyes with glaucoma and corneal disease. DESIGN: Retrospective, interventional case series. PARTICIPANTS: All patients who underwent both GDI surgery with pars plana tube insertion and PK before September 1997 were included. METHODS: The medical records of 34 consecutive patients (34 eyes) who had undergone GDI (Baerveldt, Pharmacia & Upjohn, Kalamazoo, MI; Molteno, IOP INC:, Costa Mesa, CA; Krupin, Hood Laboratories, Pembroke, MA; or Ahmed, New World Medical, Rancho Cucamonga, CA) insertion before, concurrent with, or after PK were reviewed retrospectively. All corneal grafts were clear before GDI surgery for patients who underwent glaucoma surgery after PK. Outcomes were evaluated using Kaplan-Meier life-table analysis. MAIN OUTCOME MEASURES: Clinical outcome assessment included corneal graft clarity, intraocular pressure (IOP), visual acuity, and identification of complications. RESULTS: Mean follow up after completion of both GDI surgery and PK was 12.1 +/- 8.4 months (range, 0 31.8 months). Twelve- and 24-month life-table rates for complete success after both GDI and PK were 63% and 33%, respectively. Twelve- and 24-month life-table success rates for IOP control and corneal graft clarity were 85% and 62%, and 64% and 41%, respectively. Final postoperative visual acuity was the same as or better than (> or =2 Snellen lines) the preoperative level in 29 patients (85%). One or more posterior segment complications occurred in 15 (44%) patients. CONCLUSIONS: Pars plana tube insertion of GDIs is a reasonable option for patients who have undergone PK or in whom PK is anticipated, despite the need for a complete pars plana vitrectomy. Although complications related to limbal tube placement are avoided, the incidence of posterior segment complications may be higher for pars plana insertion. The potential for enhanced corneal graft survival with pars plana versus anterior segment tube placement warrants further investigation. PMID- 11382629 TI - Central retinal vessel trunk exit and location of glaucomatous parapapillary atrophy in glaucoma. AB - OBJECTIVE: To evaluate whether the position of the central retinal vessel trunk exit on the lamina cribrosa spatially correlates with the location of parapapillary atrophy in glaucoma. DESIGN: Clinic-based, observational, cross sectional study. PATIENTS: Color stereo optic disc photographs of 95 patients with primary or secondary open-angle glaucoma and 65 healthy persons were morphometrically evaluated. The intrapapillary and parapapillary region was divided into four quadrants. We determined the position of the central retinal vessel trunk exit on the lamina cribrosa surface and measured the area of parapapillary atrophy and neuroretinal rim in the four quadrants. MAIN OUTCOME MEASURES: The area of neuroretinal rim and parapapillary atrophy and the position of the central retinal vessel trunk exit. RESULTS: Comparing measurements between opposite disc quadrants showed that beta zone of parapapillary atrophy was significantly (P < 0.05) larger and that the neuroretinal rim was significantly smaller when beta zone and neuroretinal rim were measured in the disc quadrant most distant to the central retinal vessel trunk exit, than if the beta zone and neuroretinal rim were measured in the quadrant containing the vessel trunk exit. Comparing measurements in the disc quadrants between eyes with different positions of the central retinal vessel trunk exit revealed that, in the respective disc quadrant, the beta zone was significantly larger and the neuroretinal rim was smaller in eyes with the vessel trunk exiting in the opposite disc quadrant than in eyes with the vessel trunk exit located in the respective disc quadrant where the measurements were obtained. CONCLUSIONS: Position of the central retinal vessel trunk exit on the lamina cribrosa influences the location of parapapillary atrophy in glaucoma. The longer the distance to the central retinal vessel trunk exit, the more enlarged is parapapillary atrophy and the smaller is the neuroretinal rim. This relationship agrees with the spatial relationship between glaucomatous neuroretinal rim loss and enlarged parapapillary atrophy in glaucoma. Diagnostically, it may indicate that, in eyes with an abnormal configuration of parapapillary atrophy or with an abnormal position of the central retinal vessel trunk exit, early glaucomatous rim changes should be looked for in the disc sector that is most distant to the central retinal vessel trunk exit and where parapapillary atrophy may be relatively large. PMID- 11382630 TI - Stability of corneal polarization axis measurements for scanning laser polarimetry. AB - OBJECTIVE: Corneal polarization axis (CPA) has been reported to affect retardation measurements obtained with scanning laser polarimetry. The purpose of this investigation was to evaluate the longitudinal stability of CPA measurements. DESIGN: Prospective, noncomparative case series. PARTICIPANTS: Persons with normal corneas were enrolled; eyes with less than 1 year of follow up from the initial CPA measurement were excluded. INTERVENTION: We constructed a noninvasive slit-lamp-mounted device incorporating two crossed linear polarizers and an optical retarder to measure the slow axis of corneal birefringence, or CPA. MAIN OUTCOME MEASURES: Corneal polarization axis measurements. RESULTS: Seventy-one eyes of 40 individuals (23 female, 17 male) were enrolled in this investigation (mean age, 42.9 +/- 13.6 years; range, 22-85 years). Initial CPA measurements (mean, 24.0 +/- 18.0 degrees nasally downward; range, 67 degrees downward to 13 degrees nasally upward) were strongly associated (R2 = 0.88; P < 0.0001) with repeat CPA measurements (mean, 20.9 +/- 14.6 degrees nasally downward; range, 59 degrees nasally downward to 14 degrees nasally upward). The mean change in CPA was 4.1 +/- 3.2 degrees (range, 0-13 degrees ). Corneal polarization axis stability was statistically associated with the mean (initial and repeat) CPA (R2 = 0.1; P = 0.009), but not associated with age (R2 = 0.0003; P = 0.9) or gender (R2 = 0.03; P = 0.2). CONCLUSIONS: Corneal polarization axis measurements have good 1-year stability. These data suggest that CPA should not contribute significantly to longitudinal measurements of retinal nerve fiber layer thickness obtained with scanning laser polarimetry. PMID- 11382631 TI - Stress wave amplitudes during laser surgery of the cornea. AB - PURPOSE: To determine the stress wave amplitudes generated during photoablation of the cornea using an argon fluoride excimer laser. DESIGN: Experimental study using porcine eyes. METHODS: Profiles of the stress wave amplitudes and enucleated human eyes along the axis of symmetry of porcine eyes and enucleated human eyes were measured using a miniature piezoelectric transducer. The ablation parameters, fluence, and ablation diameters were varied within the range of clinical application. MAIN OUTCOME MEASURES: Stress wave amplitudes generated during photoablation. RESULTS: The stress waves pass through a pressure focus located in the posterior lens and anterior vitreous, where amplitudes of up to 100 atm were measured with a 6-mm or larger ablation zone. Posterior to this focus, the stress wave amplitudes rapidly decrease to less than 10 atm at the retinal site. Small diameter excimer laser spots (< or =1.5 mm) produce a declining stress wave with no pressure focus at the lens and anterior vitreous. CONCLUSIONS: Stress waves may be potentially hazardous to anterior structures of the human eye, including the corneal endothelium, lens and anterior vitreous face. They peak at the lens and vitreous with a broad beam, but not with small spot laser ablation. At posterior retinal and subretinal structures, they may be considered harmless. PMID- 11382632 TI - Diffuse lamellar keratitis after laser in situ keratomileusis imaged by confocal microscopy. AB - OBJECTIVE: To report the confocal microscopic findings of two different cases of diffuse lamellar keratitis (DLK) after primary laser in situ keratomileusis (LASIK) and LASIK flap relifting. DESIGN: Observational case report of two cases. METHODS: Two cases of DLK with different clinical appearances after primary LASIK and LASIK flap relift were selected. A third case of uneventful LASIK was selected as control case. Examination with a white-light tandem slit-scanning confocal microscope was performed in addition to routinely applied slit-lamp biomicroscopy. RESULTS: In both cases, confocal microscopic examination showed infiltration of cells considered most likely to be mononuclear cells and granulocytes into the flap interface. Under treatment with topical steroids, the inflammation was reduced in both cases, to a minimum within 1 week, without loss of visual acuity. At this stage, only remnants of the granulocytelike cells, but not active inflammation, were imaged by confocal microscopic examination. CONCLUSIONS: Our findings confirm the clinical observation that DLK is an inflammation confined to the flap interface. Confocal in vivo microscopy could be a useful noninvasive tool to study the course of DLK in humans. PMID- 11382633 TI - Laser in situ keratomileusis-induced (presumed) neurotrophic epitheliopathy. AB - OBJECTIVE: To evaluate tear production, corneal topography, accuracy of refractive correction, and best spectacle-corrected visual acuity in eyes that had moderate to severe rose bengal staining develop on the flap compared with eyes with little or no staining on the flap, the first few months after laser in situ keratomileusis (LASIK). None of the eyes in this study had significant preoperative dry eye disease. DESIGN: Retrospective case control study. PARTICIPANTS: Individual eyes of 19 consecutive patients with moderate to severe punctate epithelial erosions and rose bengal staining on the flap 1 to 3 months after LASIK were compared with eyes of 19 concurrent patients who did not have punctate epithelial erosions or more than trace staining on the flap develop. METHODS: Nonparametric statistical analyses were used to compare tear secretion, corneal topographic irregularity, spherical equivalent, and visual acuity 3 and 6 months after surgery. Some eyes in both groups also had analysis of tear secretion 1 month after surgery. MAIN OUTCOME MEASURES: Schirmer's test without anesthesia, the topographic corneal irregularity measurement (CIM), the difference between attempted and achieved spherical equivalent, and the loss of best spectacle-corrected visual acuity. RESULTS: There was no difference in tear production 1, 3, or 6 months after LASIK in patients who had punctate epithelial erosions and rose bengal staining on the flap develop and those who did not. There was no significant difference in the CIM or mean accuracy of the refractive correction in the two groups, but some patients had a transient decrease in best spectacle-corrected visual acuity. Flap rose bengal staining resolved by 6 months after LASIK in most affected patients. CONCLUSIONS: LASIK-induced rose bengal staining in patients without preexisting dry eye is likely neurotrophic epitheliopathy, because there is no difference in mean tear production between patients who have significant punctate epithelial erosions and rose bengal staining develop on the flap and those who do not. The signs and symptoms of LASIK-induced (presumed) neurotrophic epitheliopathy tend to resolve approximately 6 months after surgery. This disorder tends to be more common and severe in patients with pre-existing dry eye disease. PMID- 11382634 TI - Demonstration of biofilm in infectious crystalline keratopathy using ruthenium red and electron microscopy. AB - OBJECTIVE: Bacterial biofilm formation has been implicated in the pathogenesis of infectious crystalline keratopathy. Biofilm cannot be visualized by electron microscopy without the addition of a fixative that stabilizes the polysaccharide rich bacterial extracellular matrix that surrounds the bacterial colonies in a biofilm. We used ruthenium red as a fixative to evaluate corneal biopsy specimens for the presence of bacterial biofilm in three cases of infectious crystalline keratopathy (ICK) and five cases of chronic microbial keratitis without crystalline changes. DESIGN: Case series with clinicopathologic correlation. PARTICIPANTS: Eight patients underwent corneal biopsy or therapeutic keratoplasty as part of their management for chronic unresponsive microbial keratitis. METHODS: The corneal specimens removed were trisected for microbiology, pathology, and transmission electron microscopy (TEM). The TEM specimens were fixed in 2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer with 0.05% ruthenium red. MAIN OUTCOME MEASURES: Demonstration of bacterial biofilm with TEM. RESULTS: TEM demonstrated organisms with a surrounding extracellular matrix consistent with a bacterial biofilm in the three cases of ICK but not in the five other cases of chronic microbial keratitis. CONCLUSIONS: The presence of biofilm in ICK can be demonstrated with TEM with appropriate fixation techniques that stabilize the bacterial extracellular matrix. Biofilm stains intensely with periodic acid-Schiff because of the polysaccharide-rich extracellular matrix and weakly with Gram stain because of the high proportion of nonviable organisms. Biofilm formation occurs in ICK but probably not in chronic bacterial keratitis without crystalline changes. Secretion of an extracellular matrix by bacteria to form a biofilm is a response to a nutrient-deprived environment in which growth and replication is depressed. The extracellular matrix of the biofilm may mask bacterial antigens, explaining the relative lack of inflammatory response in these infections. It may also be one of the mechanisms explaining the resistance to in vivo antimicrobial therapy when in vitro sensitivities have been proven. PMID- 11382635 TI - Treatment of conjunctival and corneal intraepithelial neoplasia with topical interferon alpha-2b. AB - OBJECTIVE: To evaluate the role of topical interferon alfa-2b (IFNalpha2b) in the treatment of conjunctival and corneal intraepithelial neoplasia (CIN). DESIGN: Noncomparative case series. PARTICIPANTS: Five patients with histologically proven CIN or recurrences of proven CIN were studied prospectively. INTERVENTION: After histologic confirmation, patients were given topical recombinant IFNalpha2b (INTRON A, Schering Plough, Kenilworth, NJ) 1 million IU/ml four times a day. Patients were continued on interferon until complete resolution of the tumor had occurred. MAIN OUTCOME MEASURES: Patients were followed clinically and photographically for evidence of tumor resolution. RESULTS: All patients had complete resolution of the CIN lesion on IFNalpha2b. The mean time to clinical resolution was 11.6 weeks (range, 4-22 weeks). The mean follow-up was 17.6 months (range, 7-28 months). One patient had a clinical recurrence of his corneal CIN 1 year after tumor resolution. This patient was retreated, resulting in clinical resolution within 6 weeks, and has been tumor free for 8 months of follow-up. PMID- 11382636 TI - Primary cataract extraction and intraocular lens implantation in penetrating ocular trauma. AB - PURPOSE: To analyze the postoperative outcome and complication rate after cataract extraction or lensectomy with primary intraocular lens (IOL) implantation for penetrating traumatic cataract. DESIGN: Retrospective, nonconsecutive, noncomparative case series. METHODS: We retrospectively reviewed the files of 21 patients who were admitted to our departments because of traumatic cataract with corneal or scleral laceration caused by penetrating trauma with or without intraocular foreign body (IOFB) from 1992 through 1997. Lens aspiration or manual extracapsular cataract extraction with primary IOL implantation was performed in all patients. Removal of an IOFB was performed in eight patients. MAIN OUTCOME MEASURES: Final visual acuity and deviation of actual refraction from emmetropia and from expected postoperative refraction. RESULTS: The mean follow-up was 20.4 months. Fourteen eyes (67%) achieved final visual acuity of 20/40 or better, 95% obtained 20/60 or better final visual acuity, and all eyes achieved 20/100 or better final visual acuity. Major causes of limited visual acuity were central corneal scar and central retinal injury. Eleven eyes (57%) experienced secondary cataract and underwent neodymium:yytrium aluminum-garnet capsulotomy. CONCLUSIONS: Primary implantation of posterior chamber lenses after penetrating ocular trauma is associated with favorable visual outcome and a low rate of postoperative complications. PMID- 11382637 TI - Visual outcome in patients with isolated autosomal dominant congenital cataract. AB - OBJECTIVE: To determine the visual outcome and surgical complication rates of patients with isolated inherited congenital cataract. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Patients and their families were ascertained from the genetic eye clinic and outpatient databases of Moorfields Eye Hospital, London, and invited to participate in the study. Four hundred twenty-two individuals from 72 pedigrees with this form of autosomal dominant cataract underwent ophthalmologic assessment. MAIN OUTCOME MEASURES: Visual acuity and surgical complications (glaucoma, retinal detachment, amblyopia). RESULTS: In this study 49.4% of patients (46.8% of those operated) achieved a visual acuity (VA) of 20/40 or better, 35.9% (36.1% of those operated) a VA between 20/50 and 20/200, and 14.7% (17.1% of those operated) worse than 20/200. Opacities that were more diffuse or did not lie close to the visual axis were associated with a better prognosis for vision; 6.6% had glaucoma and 5.0% had retinal detachment develop. CONCLUSIONS: Patients with isolated inherited congenital cataract have a better visual and surgical outcome than those with coexisting ocular and systemic abnormalities. The improved prognosis is related in part to the lack of other developmental abnormalities of the eye, and, because inherited cataracts are often partial at birth, surgery may be delayed to later infancy and childhood when there is a lower incidence of surgical complications and refractive correction is easier. Certain inherited phenotypes (lamellar, pulverulent, polymorphic, coralliform, and cortical) also seem to have a better prognosis, and this should be borne in mind when counseling these families. A large number of the patients in this study underwent surgery many years previously, when surgical outcomes were less favorable, and thus the results of this study establish only a minimum acuity dataset for the purposes of counseling. PMID- 11382638 TI - Second primary tumors in hereditary retinoblastoma: a register-based study, 1945 1997: is there an age effect on radiation-related risk? AB - OBJECTIVE: The aim of this study is to evaluate the influence of age at external beam irradiation (EBRT) on the occurrence of second primary tumors (SPTs) inside and outside the irradiation field in hereditary retinoblastoma patients. DESIGN: Cross-sectional study. PARTICIPANTS: The study included 263 hereditary retinoblastoma patients born in The Netherlands between 1945 and 1997. METHODS: A national register-based follow-up cohort study was performed on hereditary retinoblastoma patients. Information on therapy, age at irradiation, and location of SPT was obtained from the register. The Kaplan-Meier method calculated cumulative incidences of SPT in three subgroups: irradiation before (early EBRT) and after 12 months of age (late EBRT), and no irradiation. The Mantel-Cox method determined the statistical significance of differences between the cumulative incidence curves. MAIN OUTCOME MEASURES: Development of SPT inside and outside a precisely defined irradiation field in relation to age at irradiation. Our definition excluded pineoblastoma as SPT, because they constitute part of a trilateral retinoblastoma; in addition, they lie outside the field of irradiation. RESULTS: The cumulative incidence of SPT at the age of 25 years was 22% (95% confidence intervals 13%-34%) in the early EBRT group, 3% (0%-14%) in the late EBRT group, and 5% (1%-16%) in the nonirradiated group (Mantel-Cox overall: P = 0.001; between early and late EBRT, P = 0.04). However, in early irradiated patients, the incidence of SPTs inside and outside the irradiation field was similar (11%), and the difference between early and late EBRT in incidence of SPT inside the field of irradiation was less prominent than overall (11% vs. 3%: P = 0.37). Sensitivity analysis showed the results depended on the way SPT, irradiation field, and, especially, pineoblastomas are defined. CONCLUSIONS: Hereditary retinoblastoma confers an increased risk for the development of SPT, especially in patients treated with EBRT before the age of 12 months. However, the presence of similar numbers of SPTs inside and outside the irradiation field suggests that irradiation is not the cause. In other words, this study does not show an age effect on radiation-related risk. Rather, early EBRT is probably a marker for other risk factors of SPT. PMID- 11382639 TI - Basal cell carcinoma of the eyelid associated with Gorlin-Goltz syndrome. AB - PURPOSE: To describe the ophthalmic and systemic features in a series of patients initially seen with eyelid basal cell carcinoma associated with Gorlin-Goltz syndrome. DESIGN: Retrospective noncomparative case series. PARTICIPANTS: Of 105 consecutive patients with eyelid basal cell carcinoma managed at an Ocular Oncology Center between January 1973 and December 1999, four patients with Gorlin Goltz syndrome were identified. METHODS: The ophthalmic and systemic features, management, and outcome of patients with eyelid basal cell carcinoma associated with Gorlin-Goltz syndrome were analyzed. The published literature on Gorlin Goltz syndrome, specifically related to genetics, systemic features, ophthalmic associations, and prophylactic management strategies, was reviewed. MAIN OUTCOME MEASURES: Response of the eyelid basal cell carcinoma to treatment and the final systemic condition were the main outcome measures. RESULTS: All four patients had a family history of Gorlin-Goltz syndrome. The systemic manifestations included multiple basal cell carcinomas in all the patients, frontal bossing or increased occipitofrontal circumference in three patients, palmar pits in two patients, odontogenic keratocyst in one patient, ectopic calcification in one patient, and bifid rib in one patient. The mean age at the detection of the first basal cell carcinoma was 30 years (range, 16-38 years). All four patients had multiple basal cell carcinomas on the face and elsewhere. The eyelid basal cell carcinoma was advanced with orbital infiltration in three patients, one of whom opted for palliative radiotherapy, whereas the other two underwent orbital exenteration. The fourth patient, who had localized recurrent basal cell carcinoma in the upper eyelid, was treated with excision and eyelid reconstruction. At the final follow up (mean, 41 months), eyelid basal cell carcinoma was cured in three patients and stable in one patient. No patient had life-threatening sequelae of Gorlin-Goltz syndrome. CONCLUSIONS: Gorlin-Goltz syndrome is a rare autosomal dominant cancer predisposition syndrome that may be associated with eyelid basal cell carcinoma. The associated systemic findings may be a clue to the diagnosis of this condition. It is important to recognize Gorlin-Goltz syndrome when a patient has multiple basal cell carcinomas or when a young patient with eyelid basal cell carcinoma is seen by an ophthalmologist, because lifelong monitoring is essential for patient management. PMID- 11382640 TI - Sebaceous carcinoma of the eyelid associated with retinoblastoma. AB - OBJECTIVE: To analyze the association between retinoblastoma (Rb) and sebaceous carcinoma (SC) of the eyelid to improve surveillance of survivors of RB: DESIGN: Case report and systematic literature review. METHODS: Ten patients who had SC develop after Rb were identified by systematic literature review, and a child who died with lymph node, lung, and liver metastases 7 years after irradiation for Rb is described. The data were analyzed by univariate statistics, including cumulative frequency distribution plots and Kaplan-Meier analysis. RESULTS: Of 11 children with SC of the eyelid who all had hereditary RB, 9 (82%; 95% confidence interval, 48-98) received a median of 46 Gy (range, 21-89) of radiotherapy at a median age of 16 months (range, 0.5-15 years) and had SC develop within the field of radiation. Their median age at diagnosis of SC was 14 years (range, 8-30 years), median diagnostic delay 12 months (range, 6 months-3 years), and median interval from irradiation 11 years (range, 5-26 years); 7 of them (78%; 95% confidence interval, 40-97) were diagnosed between 5 and 15 years after radiotherapy. SC also developed at the age of 32 and 54 years in two nonirradiated Rb patients. Five patients had regional lymph node metastases after a median time of 12 months (range, 1 month-24 years). The 5-year cumulative probability of survival was 87%. CONCLUSIONS: SC of the eyelid may occur in patients with hereditary Rb regardless of primary treatment, especially within the field 5 to 15 years after radiotherapy. PMID- 11382641 TI - Orbital inflammatory pseudotumor and ischemic vasculitis in Churg-Strauss syndrome: report of two cases and review of the literature. AB - OBJECTIVE: To clarify the characteristics of ocular manifestations in Churg Strauss syndrome (allergic granulomatosis and angiitis). DESIGN: Two interventional case reports and literature review. PARTICIPANTS: Two patients with Churg-Strauss syndrome with ocular manifestations are described; 15 previously reported cases and the present 2 cases of Churg-Strauss syndrome with ocular manifestations are reviewed. INTERVENTION: Ocular manifestations were divided into two groups: orbital inflammatory pseudotumor and ischemic vasculitis. MAIN OUTCOME MEASURES: The onset, conjunctival involvement, orbital imaging, antineutrophil cytoplasmic antibodies (ANCA), and visual prognosis were evaluated. RESULTS: The characteristics of the orbital inflammatory pseudotumor type (eight cases) are chronic onset, positive conjunctival involvement, abnormalities in orbital imaging studies, negative ANCA, and good visual prognosis. The ischemic type (nine cases) is characterized by sudden onset, no conjunctival involvement or abnormalities in imaging studies, positive ANCA, and occasional poor visual prognosis. CONCLUSIONS: Orbital inflammatory pseudotumor and ischemic vasculitis may represent two essential characteristics of Churg Strauss syndrome, granulomatosis and angiitis, respectively. The clinical features of the two types are so distinct that differentiation may be meaningful for diagnosis and treatment of Churg-Strauss syndrome with ocular manifestations. PMID- 11382642 TI - Methotrexate therapy for chronic noninfectious uveitis: analysis of a case series of 160 patients. AB - PURPOSE: To evaluate the outcomes of patients with chronic noninfectious uveitis unresponsive to conventional antiinflammatory therapy who were treated with methotrexate. DESIGN: Retrospective noncomparative interventional case series. PARTICIPANTS: All patients with chronic noninfectious uveitis treated with methotrexate at a single institution from 1985 to 1999. METHODS: Charts of patients seen on the Ocular Immunology & Uveitis Service at the Massachusetts Eye & Ear Infirmary were reviewed. Patients with chronic uveitis of noninfectious origin treated with methotrexate were included in the study. MAIN OUTCOME MEASURES: Control of inflammation, steroid-sparing effect, visual acuity, adverse reactions. RESULTS: A total of 160 patients met the inclusion criteria. Control of inflammation was achieved in 76.2% of patients. Steroid-sparing effect was achieved in 56% of patients. Visual acuity was maintained or improved in 90% of patients. Side effects requiring discontinuation of medication occurred in 18% of patients. Potentially serious adverse reactions occurred in only 8.1% of patients. There was neither long-term morbidity nor mortality caused by methotrexate. CONCLUSIONS: Methotrexate is effective in the treatment of chronic noninfectious uveitis that fails to respond to conventional steroid treatment. It is an effective steroid-sparing immunomodulator, is a safe medication, and is well tolerated. PMID- 11382643 TI - Pars plana vitrectomy for cystoid macular edema secondary to sarcoid uveitis. AB - OBJECTIVE: To examine the results of pars plana vitrectomy for cystoid macular edema secondary to sarcoid uveitis resistant to medical treatment. DESIGN: Retrospective, interventional, noncomparative case series. SUBJECTS: Fourteen consecutive subjects (18 eyes) with cystoid macular edema associated with sarcoid uveitis resistant to medical treatment. INTERVENTION: All eyes underwent pars plana vitrectomy. Nine eyes also underwent peeling of the epiretinal membrane or removal of the posterior vitreous cortex. MAIN OUTCOME MEASURES: Status of macular edema, visual acuity, and complications. RESULTS: Ten eyes (56%) improved 2 or more lines of Snellen visual acuity within 12 months. Six eyes (33%) remained unchanged, within a line of preoperative Snellen visual acuity, and two eyes (11%) worsened by 2 or more lines of Snellen visual acuity. Slit-lamp biomicroscopy showed that cystoid macular edema had resolved in 14 eyes (78%) within 9 months postoperatively. One eye (6%) had minimal edema, whereas three eyes (17%) remained unchanged biomicroscopically at the final visit. Postoperative complications included cataract formation, glaucoma, optic nerve atrophy, epiretinal membrane formation, and tractional retinal detachment. No severe postoperative inflammation was noted. CONCLUSIONS: Pars plana vitrectomy seems to have a beneficial effect on cystoid macular edema caused by sarcoidosis resistant to medical treatment. PMID- 11382644 TI - The epidemiology of giant cell arteritis : a 12-year retrospective study. AB - OBJECTIVE: To study the incidence of biopsy-proven giant cell arteritis (GCA) in a Hispanic population with clinical features suggestive of GCA. DESIGN: Retrospective review. PARTICIPANTS: Records of 121 consecutive patients who underwent temporal artery biopsy at the Doheny Eye Institute and the Los Angeles County/University of Southern California Hospital from January 1986 through April 1998 were reviewed. MAIN OUTCOME MEASURES: The incidence of biopsy-proven GCA was determined, and the biopsy-positive group was compared with the biopsy-negative group. Study variables included age at diagnosis, gender, erythrocyte sedimentation rate (ESR), and ethnic background. RESULTS: Among these 121 patients who underwent temporal artery biopsy, the mean age of those in the biopsy-positive group (75.2 +/- 5.0 years) was higher than that of those in the biopsy-negative group (69.1 +/- 9.2 years; P < 0.0001). There was no statistical correlation between biopsy-positive and biopsy-negative groups for gender or ESR level, but ESR was statistically significant for whites when we controlled for race. Nineteen of 66 white patients (29%) had positive biopsy results, whereas only 1 of the 9 Asian patients (11%) none of the 40 Hispanic patients (0%; P < 0.0001) and none of the 6 African American patients (0%) had positive biopsy results. CONCLUSIONS: Giant cell arteritis occurs primarily in the white population. None of the Hispanic patients in our study was found to have positive biopsy results. Hispanic persons may have unknown factors that protect them from this disease. Further study is necessary to examine the genetic predisposition. PMID- 11382645 TI - Long-term follow-up of unoperated macular holes. AB - OBJECTIVE: To investigate the anatomic and visual acuity outcomes among patients with unoperated macular holes and at least 5 years of follow-up. DESIGN: Retrospective, noncomparative case series from an institutional practice setting. PARTICIPANTS: All patients with unoperated full-thickness macular holes evaluated at Bascom Palmer Eye Institute between January 1, 1968 and December 31, 1993 and observed for at least 5 years. METHODS: Demographic and clinical data were abstracted from patients' medical records and ophthalmologic photography records. For patients with bilateral macular holes, only one eye was included. MAIN OUTCOME MEASURES: Visual acuity and clinical features on initial examination, at 5 years, and at final follow-up. RESULTS: The study included 65 eyes of 65 patients with a median age of 65 years (range, 52-85 years) and a median follow up of 9.3 years (range, 5-29 years). On initial examination at Bascom Palmer Eye Institute, the macular hole was stage 2 in 15 eyes (24%), stage 3 in 23 eyes (37%), and stage 4 in 25 eyes (40%). At final follow-up, the macular hole was stage 3 in 10 eyes (16%) and stage 4 in 53 eyes (84%). Visual acuity was 20/200 or worse in 35 eyes (54%) on initial examination, in 43 eyes (74%) at 5 years, and in 53 eyes (82%) at final follow-up. Poorer visual acuity on initial examination was a significant predictor of poorer final vision (P < 0.01). Other accompanying clinical features such as the presence of operculum, posterior vitreous detachment, and epiretinal membrane were not significantly associated with final vision. Throughout follow-up, there was a redistribution and reduced number of yellow nodular opacities at the level of the retinal pigment epithelium at the base of the macular holes and the development of retinal pigment epithelial atrophy around the macular holes. CONCLUSIONS: Long-term follow-up of unoperated macular holes demonstrates progression in hole size and stage, vision loss which generally stabilizes at the 20/200 to 20/400 level, a redistribution and reduced number of yellow nodular opacities at the level of the retinal pigment epithelium, and the development of retinal pigment epithelial atrophy surrounding the macular hole, resulting in a "bull's-eye" macular appearance. PMID- 11382646 TI - Exudative retinal detachment in relapsing polychondritis : case report and literature review. AB - OBJECTIVE: To report the atypical ocular symptoms (arterialized conjunctival vessels, exudative retinal detachment) that can be the presenting manifestations of relapsing polychondritis. DESIGN: Observational case report and literature review. METHODS: A complete ocular and systemic evaluation was performed on a patient with relapsing polychondritis and exudative retinal detachment. MAIN OUTCOME MEASURES: Retinal, choroidal, and scleral findings. RESULTS: A 73-year old man with relapsing polychondritis presented with a unilateral large bullous exudative retinal detachment and marked choroidal and scleral thickening bilaterally. CONCLUSIONS: Ophthalmologists should consider relapsing polychondritis in the differential diagnosis of exudative retinal detachment. A combination of echographic and laboratory findings can assist in the accurate diagnosis of this rare condition. PMID- 11382647 TI - The performance of the refractive status and vision profile survey in a contact lens clinical trial. AB - OBJECTIVE: To evaluate the Refractive Status and Vision Profile (RSVP) questionnaire in a contact lens clinical trial and to assess the feasibility of its use in future trials. DESIGN: Randomized, crossover clinical trial and survey. PARTICIPANTS: The RSVP was administered to 50 contact lens-wearing participants recruited from a large contact lens clinic. METHODS: RSVP scores were calculated for subjects at three outcome visits: (1) baseline, (2) after wearing daily disposable contact lenses, and (3) after wearing disposable extended wear contact lenses. One-sample t tests were used to compare baseline survey scores with those previously reported for the instrument. The relation of survey scores within and between subjects was assessed with repeated measures analysis of variance. Statistical power and sample size calculations were performed to determine the number of subjects required in future studies to detect differences in RSVP subscale scores. MAIN OUTCOME MEASURES: Eight subscale scores (concern, driving, expectations, glare, optical problems, physical and social functioning, problems with corrective lenses, and symptoms) and the overall RSVP score were calculated on the basis of guidelines reported for scoring of the instrument. RESULTS: Five of eight mean subscale scores (concern, expectations, driving, optical problems, and problems with corrective lenses) and the overall score were significantly lower than those previously reported. No differences were found in the following subscales after contact lens treatment: concern, expectations, physical and social functioning, driving, optical problems, problems with corrective lenses, or the overall RSVP score. Significant differences were found on the symptoms (P = 0.03) and glare (P = 0.05) subscales; post hoc testing revealed that the significant differences in these scores occurred between baseline and daily disposable lens wear and not between treatment modalities (daily disposable and disposable extended wear). A sample size of 420 would be required to find clinically relevant significant differences (25%) for all subscales in a similar one-sample clinical trial. A more restrictive difference (10%) would require more than 1100 subjects for four of the nine subscales. CONCLUSIONS: The RSVP questionnaire may not be the appropriate quality-of-life instrument for use in a contact lens clinical trial. The instrument seems to lack sensitivity and discriminative validity in evaluating contact lens treatments. PMID- 11382648 TI - Vitamin E: too much or not enough? PMID- 11382649 TI - Energy density of foods: building a case for food intake management. PMID- 11382650 TI - Do trans fatty acids increase the incidence of type 2 diabetes? PMID- 11382651 TI - Critique of the requirement for vitamin E. AB - The Food and Nutrition Board of the Institute of Medicine recently published a comprehensive evaluation of antioxidants in human diets that includes dietary reference intakes for vitamin E. The new dietary reference intake is 15 mg (35 mol)/d for adults, which is 50% greater than the generous allowance in the 10th edition of Recommended Dietary Allowances published in 1989. Much of the data interpreted in these publications came from studies sponsored by the Committee of Nutritional Studies at Elgin State Hospital (Elgin, IL) of an earlier Food and Nutrition BOARD: The 50% increase in the recommended dietary allowances for vitamin E is not supported by any new data. It is possible that the publication of the Institute of Medicine did not take into consideration the effects of the oxidized lipids in the diets used to promote the development of vitamin E deficiency. If lipids, oxidized to remove tocopherols, had not been a part of the experimental diets, the minimum requirement for vitamin E would have been too small for possible evaluation. Studies on the different effects of saturated and oxidized lipids in the production of encephalomalacia in chicks and muscular dystrophy in rats are reviewed. The tolerable upper intake level of vitamin E supplementation is reported to be 1000 mg/d. It is possible that the universal consumption of aspirin may not have been taken into consideration when this level was determined. Vitamin E plus aspirin may increase the tendency to hemorrhage, which makes a lower upper intake level worth consideration. PMID- 11382652 TI - "Images" of nutrition in medical education and primary care. AB - In this article I describe challenges to medical-nutrition educators and an opportunity provided by the Association of American Medical Colleges Medical School Objectives Project. Brief snapshots of nutrition education are given during the family medicine residency and through continuing medical education programming. An argument is made to provide education that focuses not on knowledge, but on confidence and skill in providing nutrition services to primary care patients. Medical-nutrition educators are challenged to share curriculum ideas and to explore ways to use technology to become a more effective community. PMID- 11382653 TI - Energy density of foods affects energy intake across multiple levels of fat content in lean and obese women. AB - BACKGROUND: The results of previous studies indicated that energy density, independent of fat content, influences energy intake. In most studies, however, both fat content and energy density were lower than in typical American diets. OBJECTIVE: We examined the influence of energy density on intake when fat content was above, below, or similar to the amount of fat typically consumed and when energy density was closer to that of American diets. DESIGN: Lean (n = 19) and obese (n = 17) women consumed all meals daily in our laboratory during 6 experimental sessions. The main entrees, consumed ad libitum, were formulated to vary in fat content (25%, 35%, and 45% of energy) and energy density (5.23 kJ/g, or low energy density, and 7.32 kJ/g, or high energy density) but to have similar palatability. RESULTS: Energy density influenced energy intake across all fat contents in both lean and obese women (P < 0.0001). Women consumed less energy in the low (7531 kJ) than in the high (9414 kJ) energy density condition. Despite this 20% lower energy intake, there were only small differences in hunger (7%) and fullness (5%). Women consumed a similar volume, but not weight, of food daily across conditions. Differences in intake by weight, but not volume, occurred because for some versions of manipulated foods, weight and volume were not directly proportional. CONCLUSIONS: Energy density affected energy intake across different fat contents and at levels of energy density comparable with those in typical diets. Furthermore, our findings suggest that cues related to the amount of food consumed have a greater influence on short-term intake than does the amount of energy consumed. PMID- 11382654 TI - Dietary fat intake and risk of type 2 diabetes in women. AB - BACKGROUND: The long-term relations between specific types of dietary fat and risk of type 2 diabetes remain unclear. OBJECTIVE: Our objective was to examine the relations between dietary fat intakes and the risk of type 2 diabetes. DESIGN: We prospectively followed 84204 women aged 34-59 y with no diabetes, cardiovascular disease, or cancer in 1980. Detailed dietary information was assessed at baseline and updated in 1984, 1986, and 1990 by using validated questionnaires. Relative risks of type 2 diabetes were obtained from pooled logistic models adjusted for nondietary and dietary covariates. RESULTS: During 14 y of follow-up, 2507 incident cases of type 2 diabetes were documented. Total fat intake, compared with equivalent energy intake from carbohydrates, was not associated with risk of type 2 diabetes; for a 5% increase in total energy from fat, the relative risk (RR) was 0.98 (95% CI: 0.94, 1.02). Intakes of saturated or monounsaturated fatty acids were also not significantly associated with the risk of diabetes. However, for a 5% increase in energy from polyunsaturated fat, the RR was 0.63 (0.53, 0.76; P < 0.0001) and for a 2% increase in energy from trans fatty acids the RR was 1.39 (1.15, 1.67; P = 0.0006). We estimated that replacing 2% of energy from trans fatty acids isoenergetically with polyunsaturated fat would lead to a 40% lower risk (RR: 0.60; 95% CI: 0.48, 0.75). CONCLUSIONS: These data suggest that total fat and saturated and monounsaturated fatty acid intakes are not associated with risk of type 2 diabetes in women, but that trans fatty acids increase and polyunsaturated fatty acids reduce risk. Substituting nonhydrogenated polyunsaturated fatty acids for trans fatty acids would likely reduce the risk of type 2 diabetes substantially. PMID- 11382655 TI - Association between B vitamin intake and plasma homocysteine concentration in the general Dutch population aged 20-65 y. AB - BACKGROUND: An elevated plasma total homocysteine (tHcy) concentration is associated with an increased risk of cardiovascular diseases. Folate, riboflavin, vitamin B-6, and vitamin B-12 are essential in homocysteine metabolism. OBJECTIVE: The objective was to describe the association between dietary intakes of folate, riboflavin, vitamin B-6, and vitamin B-12 and the nonfasting plasma tHcy concentration. DESIGN: A random sample of 2435 men and women aged 20-65 y from a population-based Dutch cohort examined in 1993-1996 was analyzed cross sectionally. RESULTS: Univariately, intakes of all B vitamins were inversely related to the plasma tHcy concentration. In multivariate models, only folate intake remained inversely associated with the plasma tHcy concentration. Mean plasma tHcy concentrations (adjusted for intakes of riboflavin, vitamin B-6, vitamin B-12, and methionine and for age, smoking, and alcohol consumption) in men with low (first quintile: 161 microg/d) and high (fifth quintile: 254 microg/d) folate intakes were 15.4 and 13.2 micromol/L, respectively; in women, plasma tHcy concentrations were 13.7 and 12.4 micromol/L at folate intakes of 160 and 262 microg/d, respectively. In men, the difference in the mean plasma tHcy concentration between men with low and high folate intakes was greater in smokers than in nonsmokers (2.8 compared with 1.6 micromol/L) and greater in nondrinkers than in drinkers of >2 alcoholic drinks/d (3.5 compared with 1.4 micromol/L). In women, the association between folate intake and plasma tHcy was not modified by smoking or alcohol consumption. CONCLUSIONS: In this Dutch population, folate was the only B vitamin independently inversely associated with the plasma tHcy concentration. Changing dietary habits may substantially influence the plasma tHcy concentration in the general population. PMID- 11382656 TI - Effect of iron-fortified candies on the iron status of children aged 4-6 y in East Jakarta, Indonesia. AB - BACKGROUND: Iron deficiency anemia is the most prevalent nutrition problem in young children. One possible strategy to prevent iron deficiency anemia in this population group is the fortification of affordable food. OBJECTIVE: This study was designed to assess whether iron-fortified candies can improve iron status and are acceptable to children aged 4-6 y. DESIGN: A double-blind, placebo-controlled intervention study was conducted in Jakarta, INDONESIA: The children were randomly assigned to 1 of 2 treatment groups: a fortified group (n = 57) and a placebo group (n = 60). Every week for 12 wk, 30 g (10 pieces) candy was given to the children. The candy given to the fortified group contained 1 mg elemental Fe/g and very small amounts of other vitamins and minerals. RESULTS: The hemoglobin concentration of the fortified group increased by 10.2 g/L (95% CI: 8.3, 12 g/L) whereas that of the placebo group increased by 4.0 g/L (2.0, 6.0 g/L; P < 0.001). Anemia prevalence decreased from 50.9% at the start of the intervention to 8.8% after 12 wk of intervention in the fortified group (P < 0.001) and from 43.3% to 26.7% in the placebo group (P < 0.05). After 12 wk of intervention, the serum ferritin concentration was 71% higher than at baseline in the fortified group and 28% higher in the placebo group (P < 0.001). Acceptability of the iron-fortified candies was good. The per capita cost of the supplement was approximately US$0.96-1.20 for the 12 wk of intervention. CONCLUSION: Iron-fortified candies were effective for improving the iron status of young children and might be an affordable way to combat iron deficiency in children of low-to-middle income groups. PMID- 11382657 TI - Consumption of flavonoids in onions and black tea: lack of effect on F2 isoprostanes and autoantibodies to oxidized LDL in healthy humans. AB - BACKGROUND: Oxidative damage to lipids in vivo may be involved in the development of atherosclerosis and cancer. Onions and black tea are foods rich in flavonoids, predominantly the flavonoid quercetin, which is a potent in vitro inhibitor of membrane lipid peroxidation and LDL oxidation. OBJECTIVE: Our objective was to investigate the effects of consuming a high-flavonoid (HF) diet enriched with onions and black tea on indexes of oxidative damage in vivo compared with a low flavonoid (LF) diet. DESIGN: Thirty-two healthy humans were studied in a randomized crossover design. Indexes of oxidative damage used were plasma F2 isoprostanes (a biomarker of lipid peroxidation in vivo) and the titer of antibodies to malondialdehyde (MDA)-modified LDL. RESULTS: There were no significant differences in the intake of macronutrients or assessed micronutrients, plasma F2-isoprostane concentrations, and MDA-LDL autoantibody titer between the HF and LF dietary treatments. In the men, however, plasma concentrations of the F2-isoprostane 8-epi-prostaglandin F2alpha were slightly higher after the HF treatment phase than after the LF treatment [0.31 +/- 0.029 nmol/L (111 +/- 10.4 ng/L) compared with 0.26 +/- 0.022 nmol/L (92 +/- 7.8 ng/L); P = 0.041]. In all subjects, plasma quercetin concentrations were significantly higher after the HF treatment phase than after the LF treatment: 221.6 +/- 37.4 nmol/L compared with less than the limit of detection of 66.2 nmol/L. CONCLUSION: Flavonoid consumption in onions and tea had no significant effect on plasma F2 isoprostane concentrations and MDA-LDL autoantibody titer in this study and thus does not seem to inhibit lipid peroxidation in humans. PMID- 11382658 TI - Zinc supplementation might potentiate the effect of vitamin A in restoring night vision in pregnant Nepalese women. AB - BACKGROUND: Zinc deficiency may result in abnormal dark adaptation or night blindness, a symptom primarily of vitamin A deficiency. During a placebo controlled trial in Nepal, weekly vitamin A supplementation of women reduced but failed to eliminate the incidence of night blindness during pregnancy, suggesting a role for zinc. OBJECTIVE: The study examined the efficacy of daily zinc supplementation in restoring night vision of pregnant women who developed night blindness while routinely receiving either vitamin A, beta-carotene, or placebo in a field trial. DESIGN: Women (n = 202) who reported to be night blind during pregnancy were randomly assigned in a double-blind manner, stratified on vitamin A, beta-carotene, or placebo receipt, to receive 25 mg Zn or placebo daily for 3 wk. Thus, the 6 groups studied were as follows: beta-carotene + zinc, beta carotene alone, vitamin A + zinc, vitamin A alone (vitamin A + placebo), zinc alone (zinc + placebo), and placebo (2 placebos: one for the vitamin A or beta carotene study and one for the zinc study). Women underwent a clinic-based assessment that included pupillary threshold testing and phlebotomy before and after supplementation. Supplement use and daily history of night blindness were obtained at home twice every week. RESULTS: Zinc treatment increased serum zinc concentrations, but alone (zinc alone group), failed to restore night vision or to improve dark adaptation. However, women in the vitamin A + zinc group who had baseline serum zinc concentrations <9.9 micromol/L were 4 times more likely to have their night vision restored (95% CI: 1.1, 17.3) than were women in the placebo group and tended to have a small improvement in pupillary threshold scores (by 0.21 log candela/m2; P = 0.09). CONCLUSION: These data suggest that zinc potentiated the effect of vitamin A in restoring night vision among night blind pregnant women with low initial serum zinc concentrations. PMID- 11382659 TI - Chronic administration of pharmacologic doses of vitamin E improves the cardiac autonomic nervous system in patients with type 2 diabetes. AB - BACKGROUND: Type 2 diabetes is associated with elevated oxidative stress and declines in antioxidant defense. The disease is also characterized by an imbalance in the ratio of cardiac sympathetic to parasympathetic tone. Antioxidants, vitamin E in particular, may have beneficial effects on the cardiac autonomic nervous system through a decline in oxidative stress. OBJECTIVE: We investigated the possible effects of vitamin E on the cardiac autonomic nervous system, as assessed by analysis of heart rate variability, in patients with type 2 diabetes and cardiac autonomic neuropathy. DESIGN: In a double-blind randomized controlled trial, 50 patients with type 2 diabetes were assigned to treatment with vitamin E (600 mg/d) or placebo for 4 mo. RESULTS: The anthropometric characteristics of the patients remained unchanged throughout the study. Chronic vitamin E administration was associated with decreases in concentrations of glycated hemoglobin (P < 0.05), plasma insulin (P < 0.05), norepinephrine (P < 0.03), and epinephrine (P < 0.02); a lower homeostasis model assessment index (P < 0.05); and improved indexes of oxidative stress. Furthermore, vitamin E administration was associated with increases in the R-R interval (P < 0.05), total power (P < 0.05), and the high-frequency component of heart rate variability (HF; P < 0.05) and decreases in the low-frequency component (LF; P < 0.05) and the ratio of LF to HF (P < 0.05). Finally, change in the plasma vitamin E concentration was correlated with change in the LF-HF ratio (r = -0.43, P < 0.04) independently of changes in the homeostasis model assessment index and plasma catecholamines concentrations. CONCLUSIONS: Chronic vitamin E administration improves the ratio of cardiac sympathetic to parasympathetic tone in patients with type 2 diabetes. Such an effect might be mediated by a decline in oxidative stress. PMID- 11382660 TI - HIV and other predictors of serum beta-carotene and retinol in pregnancy: a cross sectional study in Zimbabwe. AB - BACKGROUND: Vitamin A status during pregnancy is important to maternal and infant health. OBJECTIVE: Our goal was to identify predictors of serum beta-carotene and retinol. DESIGN: This was a cross-sectional study of 1669 women (22-35 wk of gestation) in Harare, Zimbabwe, who were receiving prenatal care. The statistical effects of age, season, gestational age, gravidity, HIV-1 infection, malaria parasitemia, and serum alpha1-antichymotrypsin (ACT) on serum beta-carotene (log10 transformed) and retinol were estimated by using multiple linear regression analyses. RESULTS: HIV infection was found in 31.5% of the women; 0.4% had malaria. Serum beta-carotene concentrations (geometric x: 0.19 micromol/L) were lower in HIV-infected women than in uninfected women (10beta = 0.78; 95% CI: 0.72, 0.84) and increased with age (10beta = 1.05; 1.02, 1.07) in gravida 1 but not in gravida > or =2 (P for interaction = 0.00002). Serum retinol (x: 0.92 micromol/L) increased with age (beta = 0.004; 0.0001, 0.008) in uninfected women but not in HIV-infected women (P for interaction = 0.02) and was 0.05-micromol/L (0.02, 0.09) lower in HIV-infected women than in uninfected women at 24 y of age. Furthermore, gestational age, season, use of prenatal supplements, and malaria were predictors of serum beta-carotene. Serum retinol was lower in women carrying male (beta = -0.04; -0.08, -0.00005) and multiple (beta = -0.21; -0.35, -0.08) fetuses. Serum ACT concentrations of 0.3-0.4, 0.4-0.5, and >0.5 g/L were associated with 3%, 11%, and 44% lower serum beta-carotene and 0.04-, 0.15-, and 0.41-micromol/L lower serum retinol. Serum ACT (g/L) was higher in women with malaria than in those without (beta = 0.10; 0.03, 0.16) and in gravida 1 than in gravida > or =2 (beta = 0.012; 0.003, 0.021), but was not higher in HIV-infected women than in uninfected women (beta = 0.001; -0.008, 0.011). CONCLUSIONS: HIV infection, malaria, gravidity, and gestational age were predictors of serum beta carotene and retinol. Serum ACT was an important predictor of both and was associated with gravidity and gestational age. PMID- 11382661 TI - HIV and other predictors of serum folate, serum ferritin, and hemoglobin in pregnancy: a cross-sectional study in Zimbabwe. AB - BACKGROUND: Folate and iron status and hemoglobin concentrations are important to maternal and infant health. OBJECTIVE: Our goal was to identify predictors of serum folate, serum ferritin, and hemoglobin. DESIGN: This was a cross-sectional study of 1669 pregnant women (22-35 wk of gestation) in Harare, Zimbabwe, who were receiving prenatal care. The statistical effects of age, season, gestational age, gravidity, HIV-1 infection, malaria parasitemia, and serum alpha1 antichymotrypsin (ACT) on serum folate, serum ferritin (log10 transformed), and hemoglobin were estimated by using multiple linear regression analyses. RESULTS: Serum folate (x: 11.4 micromol/L) was 0.52-nmol/L (95% CI: 0.04, 1.0) lower in HIV-infected women than in uninfected women and 0.65-nmol/L (0.014, 1.28) lower in weeks 25-35 than in weeks 22-25. Serum ferritin (geometric x: 11.6 microg/L) was 0.93 times (0.86, 0.99) lower in HIV-infected women and 2.25 times (1.41, 3.61) higher in women with malaria parasitemia than in uninfected women. Similarly, serum ferritin was 0.71 times (0.63, 0.79) higher in weeks 32-35 than in weeks 22-25 and 1.21 times (1.12, 1.29) higher in gravida > or =3 than in gravida 1. Elevated serum ACT was a strong predictor of serum folate, serum ferritin, and hemoglobin. HIV infection was associated with a 12.9-g/L (8.9, 16.8) lower hemoglobin concentration in women with nondepleted iron stores but low serum retinol and a 7-8-g/L lower hemoglobin concentration in women with other combinations of serum ferritin and retinol (P for interaction = 0.038). Season, age, gestational age, and gravidity were not significant predictors of hemoglobin. Low serum folate, ferritin, and retinol were associated with low hemoglobin. CONCLUSIONS: HIV was associated with lower serum folate, serum ferritin, and hemoglobin. HIV infection was also associated with lower hemoglobin, particularly in women with stored iron and low serum retinol. Low serum folate, ferritin, and retinol were associated with low hemoglobin. PMID- 11382662 TI - Comparison of the peripartum and postpartum phospholipid polyunsaturated fatty acid profiles of lactating and nonlactating women. AB - BACKGROUND: Pregnancy is associated with increased absolute amounts of docosahexaenoic acid (DHA; 22:6n-3) in plasma phospholipids. Expressed as a proportion of total fatty acids, DHA declines slightly in late pregnancy but little information is available on the normalization of DHA postpartum, which may be different in lactating and nonlactating women. OBJECTIVE: The aim was to investigate maternal plasma and erythrocyte long-chain polyunsaturated fatty acids (long-chain polyenes; LCPs) postpartum, particularly DHA, in relation to lactation and dietary LCP intake. DESIGN: Healthy pregnant women who intended to breast-feed or exclusively bottle-feed their infants were studied at 36-37 wk of pregnancy. Blood samples were collected at entry, after parturition on days 2 and 5, and 1, 2, 4, 8, 16, 32, and 64 wk postpartum. Fatty acid profiles were analyzed in plasma and erythrocyte phospholipids. Dietary intakes were assessed 4 and 32 wk postpartum with a validated food-frequency questionnaire. RESULTS: After delivery, the percentages of plasma linoleic, arachidonic, eicosapentaenoic, and docosapentaenoic acids increased over time, whereas the percentage of docosapentaenoic acid decreased; the patterns of change did not differ significantly between the lactating and nonlactating groups. The percentage of DHA in plasma and erythrocyte phospholipid fatty acids declined significantly in the 2 groups, more so in the lactating women, and was enhanced when the lactation period was extended. Despite the apparent higher dietary intake of essential fatty acids in the lactating group at week 4, it was not significantly different from that of the nonlactating group. CONCLUSION: Normalization of maternal plasma and erythrocyte phospholipid n-3 LCPs differs significantly between lactating and nonlactating women postpartum but that of n-6 LCPs does not. PMID- 11382663 TI - Flavor variety enhances food acceptance in formula-fed infants. AB - BACKGROUND: Research in humans and animal models suggests that acceptance of solid foods by infants during weaning is enhanced by early experiences with flavor variety. OBJECTIVE: We tested the hypotheses that the acceptance of novel foods by formula-fed infants could be facilitated by providing the infants with a variety of flavors at the time when beikost is first introduced and that, contrary to medical lore, infants who had previously consumed fruit would be less likely to reject vegetables when first introduced than would infants without such an experience. DESIGN: The infants' acceptance of a novel vegetable (pureed carrot) and a novel meat (pureed chicken) was evaluated after a 9-d exposure period in 3 groups of infants, some of whom had previously consumed fruit. During the home-exposure period, one group was fed only carrots, the target vegetable; a second group was fed only potatoes, a vegetable that differed in flavor from carrots; and a third group was fed a variety of vegetables that did not include carrots. RESULTS: Infants fed either carrots or a variety of vegetables, but not those fed potatoes, ate significantly more of the carrots after the exposure period. Exposure to a variety of vegetables also facilitated the acceptance of the novel food, pureed chicken, and daily experience with fruit enhanced the infants' initial acceptance of carrots. CONCLUSION: These findings are the first experimental evidence to indicate that exposure to a variety of flavors enhances acceptance of novel foods in human infants. PMID- 11382664 TI - Prevalence of overweight in US children: comparison of US growth charts from the Centers for Disease Control and Prevention with other reference values for body mass index. AB - BACKGROUND: Several different sets of reference body mass index (BMI) values are available to define overweight in children. OBJECTIVE: The objective of this study was to compare the prevalence of overweight in US children calculated with 3 sets of reference BMI values: the revised growth charts of the Centers for Disease Control and Prevention (CDC-US growth charts), international standards proposed by Cole et al, and values developed by Must et al. DESIGN: Data for children and adolescents came from cross-sectional nationally representative US surveys: cycles II and III of the National Health Examination Survey (1963-1965 and 1966-1970) and the first, second, and third National Health and Nutrition Examination Surveys: NHANES I (1971-1974), II (1976-1980), and III (1988-1994). The reference values of Cole et al equivalent to a BMI of 25 were compared with the 85th percentiles from the other 2 methods; the values equivalent to a BMI of 30 were compared with the 95th percentiles. RESULTS: The 3 methods gave similar but not identical results. The reference values of Cole et al gave lower estimates than did the CDC-US growth charts for young children but higher estimates for older children. The reference values of Must et al gave much higher prevalences for younger girls than did the other 2 methods. CONCLUSIONS: Differences between methods were related to differences in data sets, smoothing methods, and theoretical approaches. All 3 methods are based on statistical criteria and incorporate arbitrary assumptions. These methods should be used cautiously, with awareness of the possible limitations. PMID- 11382665 TI - Determinants of serum enterolactone concentration. AB - BACKGROUND: The lignan enterolactone, which is produced by the intestinal microflora from dietary precursors, may protect against hormone-dependent cancers and cardiovascular diseases. OBJECTIVE: We examined the cross-sectional associations between the serum enterolactone concentration and variables related to diet and health in Finnish adults. DESIGN: Serum enterolactone was measured by using time-resolved fluoroimmunoassay in 2380 Finnish men and women aged 25-64 y who were participating in a cross-sectional national survey in 1997. Background information was collected with self-administered questionnaires and the diet was assessed with a food-frequency questionnaire. RESULTS: The median serum enterolactone concentration was 13.8 nmol/L (range: 0-95.6 nmol/L) in men and 16.6 nmol/L (range: 0-182.6 nmol/L) in women. Multiple regression analyses showed positive associations in men between the serum enterolactone concentration and constipation, consumption of whole-grain products, and intake of fruit and berries. In women, the serum enterolactone concentration was positively and independently associated with consumption of vegetables, subject age, and constipation and was negatively associated with smoking. Furthermore, female subjects of normal weight had significantly higher serum enterolactone concentrations than did their underweight or obese peers. CONCLUSIONS: The serum enterolactone concentration varies widely in the population. Of the variables we examined, the most important determinants of the serum enterolactone concentration were consumption of lignan-containing foods and constipation; however, these appeared to explain only a small part of the variation. Therefore, the role of gut microflora in the metabolism of lignans might be very important. Further studies will also be needed to determine the bioavailability and absorption rate of lignans. PMID- 11382666 TI - Variability in commercial ginseng products: an analysis of 25 preparations. AB - BACKGROUND: Because dietary supplements are not subject to the same regulations that pharmaceuticals are, there is concern among medical professionals that these products may lack purity or potency. OBJECTIVE: To determine the variability in a range of ginseng herbal products available in the United States, we identified and measured the concentration of marker compounds by using HPLC and liquid chromatography-tandem mass spectrometry. DESIGN: Twenty-five commercial ginseng preparations from the genera Panax or Eleutherococcus were obtained from a local health food store and analyzed for 7 ginsenosides (marker compounds for Panax species, which include Asian and American ginseng) and 2 eleutherosides (marker compounds for Eleutherococcus senticosus, also known as Siberian ginseng). RESULTS: All plant products were correctly identified by botanical plant species (ie, Panax species or E. senticosus); however, concentrations of marker compounds differed significantly from labeled amounts. There was also significant product to-product variability: concentrations of ginsenosides varied by 15- and 36-fold in capsules and liquids, respectively, and concentrations of eleutherosides varied by 43- and 200-fold in capsules and liquids, respectively. Although a systematic search for adulterants was not conducted, review of the HPLC and liquid chromatography-tandem mass spectrometry data suggest that no substances other than ginsenosides or eleutherosides were extracted from the plant material. CONCLUSION: Our data suggest that US ginseng products are correctly labeled as to plant genus; however, variability in concentrations of marker compounds suggests that standardization may be necessary for quality assurance and that characterization of herbal products should be considered in the design and evaluation of studies on herbal products. PMID- 11382667 TI - Effect of an integrated nutrition curriculum on medical education, student clinical performance, and student perception of medical-nutrition training. AB - BACKGROUND: Ninety-eight percent of medical schools report nutrition as a component of medical education. However, most schools do not have an identifiable nutrition curriculum. Medical schools that do include nutrition have not evaluated its effect on clinical skills. OBJECTIVE: The objective was to determine the efficacy of an integrated undergraduate medical curriculum to increase the quantity of nutrition instruction and to advance nutrition clinical skills demonstrated by medical students. DESIGN: A quasiexperimental design was constructed to determine whether an integrated nutrition curriculum increased the performance on nutrition-oriented clinical examinations of medical school classes that received 1, 2, or 3 y of the curriculum. The evaluation of the curriculum focused on 3 areas: 1) hours of nutrition instruction, 2) the application of nutrition within a clinical setting, and 3) perceptions about the nutrition curriculum. The Objective Structured Clinical Examination (OSCE) nutrition score was compared between graduating classes by use of analysis of variance. Data from the American Association of Medical Colleges were analyzed to determine the change in the proportion of students who reported that the amount of time devoted to nutrition was adequate. RESULTS: The implementation of the integrated nutrition curriculum resulted in a doubling of the total hours of required instruction in the medical curriculum (35 compared with 75 h). The mean (+/-1 SEM) OSCE nutrition score significantly improved after the implementation of the curriculum (41.7 +/- 0.9% compared with 50.6 +/- 1.1%) and the percentage of students who reported that the amount of nutrition taught during medical school was inadequate decreased (68.4% compared with 11.5%). CONCLUSION: Medical students improved their clinical nutrition practice skills through participation in an integrated nutrition curriculum. PMID- 11382669 TI - Vitamin E, cancer, and apoptosis. PMID- 11382678 TI - John P. Peters and nephrology. PMID- 11382679 TI - Composition of nephritic factor-generated glomerular deposits in membranoproliferative glomerulonephritis type 2. AB - Two observations suggest that nephritic factors (NFs) may be nephritogenic. First, glomerulonephritis is present in unusual frequency in three conditions in which the function of factor H is blocked, a dysfunction also produced by NFS: Second, in membranoproliferative glomerulonephritis (MPGN) type 2, subepithelial deposits on the paramesangial portion of the glomerular basement membrane are found only in renal biopsy specimens obtained during hypocomplementemia when NF is presumably present. In the present study, the composition of these deposits with respect to C3 derivatives was assessed by immunohistological evaluation using anti-C3c and anti-C3d. The assessment used routinely obtained photomicrographs, as well as immunohistologic examination of freshly cut tissue using the double-antibody method. Deposits in patients with typical hypocomplementemic MPGN type 2 reacted only with anti-C3c, whereas those in two patients with rapidly progressive MPGN type 2, six patients with poststreptococcal acute glomerulonephritis, and five patients with juvenile acute nonproliferative glomerulitis reacted with anti-C3d, as well as anti-C3c. Because all products derived from the breakdown of C3 except C3c react with anti-C3d, the deposits in typical MPGN type 2 must be composed only of C3c. With complete breakdown of bound C3b, C3c is released into the fluid phase. Therefore, the C3c in the deposits cannot be a product of a glomerular complement reaction, but instead must be formed in the circulation by the reaction of NF with native C3. Supporting C3c as the only constituent of these deposits is the observation that they are devoid of properdin and C5 is present in only small amounts. PMID- 11382680 TI - Improved strategy for molecular genetic diagnostics in juvenile nephronophthisis. AB - Juvenile or type 1 nephronophthisis (NPH1), an autosomal recessive cystic kidney disease, represents the most common genetic cause of end-stage renal disease in the first two decades of life. Because the disease is caused by large homozygous deletions of the NPHP1 gene in approximately 66% of patients with nephronophthisis, molecular genetic testing offers a method for the definite diagnosis of NPH1 and avoids the invasive procedure of renal biopsy. We recently developed an algorithm for molecular genetic diagnosis of NPH1 that efficiently detects homozygous deletions. However, a major limitation remained for the detection of heterozygous deletions that cause NPH1 in combination with point mutations at the other NPHP1 allele. Because a partial sequence from the NPHP1 region recently became available through the Human Genome Projects, we exploited this information to develop novel polymorphic markers from this genetic region for the detection of heterozygous deletions of NPHP1, thus bridging the diagnostic gap. Five novel polymorphic microsatellites positioned within the large common NPHP1 deletion were generated. Two multiplex polymerase chain reaction sets using two and three polymorphic markers from the NPHP1 deletion region together with one positive control marker allowed four different diagnostic problems to be solved in one diagnostic setup: (1) detection of the classic homozygous deletion of NPH1, (2) detection of a rare smaller homozygous deletion of NPH1, (3) testing for a heterozygous deletion, and (4) potential exclusion of linkage to NPHP1. The newly generated multiplex marker sets will greatly enhance the efficacy of molecular diagnostics in NPH through improved detection of heterozygous deletions. PMID- 11382681 TI - Contributory metabolic factors in the development of nephrolithiasis in patients with medullary sponge kidney. AB - Whether specific metabolic abnormalities are related to nephrolithiasis in patients with medullary sponge kidney (MSK) remains a debated issue. The purpose of this study is to determine metabolic disorders in patients with MSK and nephrolithiasis compared with idiopathic calcium-stone-forming patients. One hundred eighty-four patients with recurrent calcium-stone formations were investigated with regard to metabolic abnormalities. Of these, 22 patients (11.9%; 13 men, 9 women) showed MSK by radiological examination. MSK was defined as a kidney that presented at least three linear or round papillary opacities in the affected papilla on urography. Multiple stones (more than five) existed in both kidneys in all patients with MSK. The remaining 162 patients (109 men, 53 women) were idiopathic calcium-stone formers. Frequencies of low urine volume (urine < 1,500 mL/24 h) and hyperoxaluria (oxalate > 40 mg/24 h) were similar between the groups. Hypercalciuria (men, calcium > 300 mg/24 h; women, calcium of 250 mg/24 h) was found less frequently in the MSK group. The frequency of hypocitraturia (citrate < 300 mg/24 h) was significantly greater in the MSK group than the idiopathic group (77.3% versus 33.9%, respectively). Mean 24-hour urinary excretions of calcium, citrate, uric acid, and magnesium were significantly less in the MSK group. No differences were found in serum calcium, phosphate, and parathyroid hormone levels between the groups. Low urinary excretions of citrate and magnesium are the most typical metabolic disorders that distinguish MSK stone patients from idiopathic calcium-stone-forming patients. In addition to such anatomic abnormalities as ectatic collecting ducts, low levels of urinary inhibitors of stones seem to contribute to the pathogenesis of nephrolithiasis in patients with MSK. PMID- 11382682 TI - Transjugular renal biopsy in patients with liver disease. AB - Although transjugular renal biopsy has been used extensively in Europe, experience with its use in the United States has been limited. We report 25 patients who underwent both transjugular liver and renal biopsies in the same sitting and 4 patients who underwent only a transjugular renal biopsy. All 29 patients had both liver disease and renal abnormalities. Each patient was also believed to have a relative or absolute contraindication to a percutaneous renal biopsy (usually in the form of a bleeding abnormality). Transjugular renal biopsy yielded a quantity of tissue sufficient for diagnosis in all but 1 patient. The mean number of glomeruli obtained per biopsy was 19.4 +/- 12.2 (SD). Pathological diagnoses found were tubular injury in 5 patients, membranoproliferative glomerulonephritis in 5 patients, nephrosclerosis in 3 patients, diabetic nephropathy in 2 patients, immunoglobulin A (IgA) nephropathy in 2 patients, minimal change disease in 2 patients, end-stage renal disease in 2 patients, nonspecific changes in 1 patient, early glomerulosclerosis in 1 patient, tubular atrophy only in 1 patient, and normal renal histological characteristics in 4 patients. One patient with suspected IgA nephropathy had no histological diagnosis established because of a lack of glomeruli in the biopsy specimen. There were no instances of major bleeding from the perirenal area; however, a small perirenal hematoma was identified in 3 patients by postbiopsy computed tomography or sonography. Thus, based on our experience, transjugular renal biopsy appears to be a safe and effective procedure for establishing a histological diagnosis and is an attractive alternative biopsy method for patients with advanced liver disease and contraindications to conventional percutaneous renal biopsy. PMID- 11382683 TI - Relationship between underlying renal disease and renal transplantation outcome. AB - The purpose of this study is to better characterize graft and patient survival posttransplantation by examining survival according to underlying renal disease for all first-time renal allograft recipients in the United Network for Organ Sharing (UNOS) registry. From 1987 through 1996, the UNOS registry collected data on 23,838 living and 67,183 cadaveric renal transplantations. This investigation included all patients undergoing their first renal transplantation for whom the underlying cause of renal failure could be identified and categorized. Gross 1- and 3-year patient and graft survival according to underlying renal disease are included. In addition, a Cox proportional hazards model was created to analyze the effect of underlying disease on graft and patient survival after adjusting for comorbid conditions, demographics, and type of renal transplant (living versus cadaveric). The association between underlying disease and graft and patient survival is shown. Amyloidosis, sickle cell anemia, scleroderma, and radiation nephritis are associated with poor graft and patient survival. The risk ratio for patient mortality was more than twice that for immunoglobulin A nephropathy for a number of conditions, including analgesic nephropathy, amyloidosis, and both forms of diabetes mellitus. PMID- 11382684 TI - Lack of long-term benefits of steroid withdrawal in renal transplant recipients. AB - Glucocorticoids used in renal transplantation have been associated with numerous adverse effects. Most studies that showed short-term benefits of steroid withdrawal made comparisons for patients administered prednisone, 10 to 17.5 mg/d, versus no prednisone. Few have studied long-term benefits of steroid withdrawal. We performed a retrospective review and identified 58 patients administered cyclosporine, azathioprine, and prednisone who underwent complete steroid withdrawal. Post-steroid withdrawal follow-up was 7.6 +/- 1.9 years. Nine patients restarted prednisone therapy, 3 patients lost their grafts (2 of those restarted on prednisone therapy), and 2 patients died with functioning grafts. When prednisone dosage was tapered from 10 mg/d to 10 mg every other day, clinically significant improvements were seen in weight, systolic and diastolic blood pressure, blood pressure medications, glycosylated hemoglobin level, and diabetic medications. No further benefits were seen in these parameters and total cholesterol level on complete steroid withdrawal from prednisone, 10 mg every other day. Most of the earlier benefits were not sustained on long-term follow up, and the increase in these parameters was similar to that of a similar matched control group (that underwent transplantation during the same period) maintained on prednisone, 5 mg/d. Major differences were decreases in creatinine clearances and hemoglobin levels, which were greater in the steroid-withdrawal group (7.4 +/ 1.9 mL/min and 1.2 +/- 0.2 g/dL, respectively) compared with the control group (2.6 +/- 1.5 mL/min and 0.5 +/- 0.2 g/dL, respectively). In conclusion, most of the metabolic benefits were seen with steroid dosage taper from prednisone, 10 mg/d to 10 mg every other day, with no further benefits with steroid withdrawal. Most of these benefits were not sustained on long-term follow-up, questioning the utility of steroid withdrawal. PMID- 11382685 TI - A comparison between office and ambulatory blood pressure measurements in renal transplant patients with chronic transplant nephropathy. AB - Hypertension is an important risk factor for chronic transplant nephropathy. Therapy is usually based on casual office blood pressure (BP) measurements. However, it is not well known how casual BP predicts 24-hour BP in this population. The main focus of this study is to compare casual office BP with 24 hour ambulatory BP monitoring in renal transplant recipients with signs of chronic transplant nephropathy. Moreover, in this group, the day-night BP profile was assessed. In 36 renal transplant recipients with incipient or progressive proteinuria or an increase in serum creatinine level greater than 20%, 24-hour ambulatory BP was performed. Patients were defined as a nondipper if the mean BP decreased by less than 10% during the nighttime period. The correlation between single office and 24-hour ambulatory BPs was 0.61 for systolic BP and 0.55 for diastolic BP (P < 0.001). The mean difference between 24-hour ambulatory and single office BPs was -4.2 +/- 18.6 mm Hg (range, -44 to 36 mm Hg) for systolic BP and -1.1 +/- 10.7 mm Hg (range, -34 to 27 mm Hg) for diastolic BP; 94.5% of patients were classified as nondippers. There was a significant relation between the nightly decline in mean arterial pressure and calculated creatinine clearance (r = 0.34; P < 0.05). In conclusion, in renal transplant recipients with chronic transplant nephropathy, a large difference between office and ambulatory BPs is present, with both overestimation and underestimation of 24-hour BP by office BP measurements. Moreover, a severely disturbed day-night BP rhythm was observed. In transplant recipients with compromised graft function, office BP may not reflect 24-hour BP adequately, and ambulatory BP measurements should be considered. PMID- 11382686 TI - Prevalence and characteristics of individuals with chronic kidney disease in a large health maintenance organization. AB - This study is designed to estimate the prevalence of and gain further insight into the characteristics of the chronic kidney disease (CKD) population in a large US health maintenance organization (HMO) to better understand the CKD population in the United States overall. Analyses were performed using data from a staff and network model HMO in the southwestern United States with more than 150,000 members per year during 1994 to 1997. The estimated prevalence of CKD in the HMO population varied from 0.4% to 7.1%, depending on the definition of CKD used. Regardless of the definition, CKD was more common in men compared with women and in patients with diabetes mellitus and/or hypertension. Applying the age- and sex-specific prevalence rates in the HMO to the US population in 1990, we estimate there were approximately 9.1 million Americans with at least one elevated sex-specific creatinine (Cr) value and approximately 4.2 million Americans with at least two elevated Cr values separated by 90 days or greater, a more rigorous definition of CKD. From these results, it is apparent that there are a large number of patients in the United States with CKD. Most have not been identified because screening for CKD generally is not performed. Considering the high prevalence of CKD and the high cost and clinical morbidity associated with end-stage renal disease (ESRD), it is clear that CKD is an important public health problem. Early identification of patients with CKD would allow treatment that could slow the progression to ESRD, improve clinical outcomes, and constrain the growth of costs in the ESRD program. The time has come for a structured public and professional educational program to address this serious condition. PMID- 11382687 TI - Incidence trends and mortality in end-stage renal disease attributed to renovascular disease in the United States. AB - End-stage renal disease (ESRD) attributed to renovascular disease (RVD-ESRD) has been incompletely characterized. We determined incidence trends, clinical features, prior treatment, and survival of patients with RVD-ESRD using the US Renal Data System database. Primary causes of ESRD were assessed in patients starting ESRD therapy during 1991 to 1997. The incidence of RVD-ESRD increased from 2.9/10(6) per year (1.4% of new ESRD cases) to 6.1/10(6) per year (2.1%). The annualized increase was 12.4% per year. This is a greater rate of increase than for ESRD from diabetes mellitis (DM-ESRD; 8.3% per year) and ESRD overall (5.4% per year). The risk for RVD-ESRD versus other-cause ESRD correlated positively with age (odds ratio [OR], 1.7 per 10-year increment; P < 0.0001) and male sex (OR, 1.2; P < 0.0001) and negatively with black (OR, 0.17; P < 0.0001), Asian (OR, 0.29; P < 0.0001), and Native American race (OR, 0.31; P < 0.0001). The unadjusted prevalence of coronary heart disease, cerebrovascular disease, and peripheral vascular disease was greater in patients with RVD-ESRD versus other cause ESRD (P < 0.001). Of patients with RVD-ESRD, 5% underwent revascularization in the 2 years before ESRD compared with 0.5% of patients with other-cause ESRD, including DM-ESRD. Adjusted for age, race, sex, comorbidity, and laboratory values, the survival of patients with RVD-ESRD was similar to that for patients with other-cause ESRD (risk ratio, 1.01; P = 0.5). These findings suggest that RVD-ESRD is increasing faster than other-cause ESRD and is not independently associated with an increased mortality risk. Strategies may exist to prevent progression to ESRD and merit priority for further study. PMID- 11382688 TI - Determinants of mortality after myocardial infarction in patients with advanced renal dysfunction. AB - Previous studies using administrative data have shown high mortality in patients with renal failure requiring dialysis after acute myocardial infarction (AMI). There has been little investigation into the mortality after AMI in those with advanced renal disease who are not on dialysis therapy. We analyzed a prospective coronary care unit registry of 1,724 patients with ST segment elevation myocardial infarction admitted over an 8-year period at a single tertiary-care center. Those not on chronic dialysis therapy were stratified into groups based on corrected creatinine clearance, with cutoff values of 46.2, 63.1, and 81.5 mL/min/72 kg. Dialysis patients (n = 47) were considered as a fifth comparison group. Older age, black race, diabetes, hypertension, previous coronary disease, and heart failure were incrementally more common across increasing renal dysfunction strata. There were also graded increases in the relative risk for atrial and ventricular arrhythmias, heart block, asystole, development of pulmonary congestion, acute mitral regurgitation, and cardiogenic shock. Primary angioplasty, thrombolysis, and beta-blockers were used less often across the risk strata (P < 0.0001 for all trends). There was an early mortality hazard (age adjusted relative risk, 8.76; P < 0.0001) for those with renal dysfunction but not on dialysis therapy for the first 60 months, followed by graded decrements in survival across increasing renal dysfunction strata. The excess mortality in this population appears to be mediated through arrhythmias, adverse hemodynamic events, and the lower use of mortality-reducing therapy. PMID- 11382689 TI - Plasma brain natriuretic peptide levels in chronic hemodialysis patients: influence of coronary artery disease. AB - A noninvasive biochemical testing method for early detection and monitoring the condition of cardiac complications in hemodialysis (HD) patients would be useful and might lead to improved survival. The aim of this study is to clarify the pathophysiological significance of plasma brain natriuretic peptide (BNP) levels in HD patients with and without coronary artery disease (CAD). We measured plasma atrial natriuretic peptide (ANP) and BNP levels on Monday, Wednesday, and Friday before and after HD in 28 consecutive patients who underwent HD three times weekly. In addition, we measured plasma ANP and BNP levels in 21 HD patients with CAD and 27 HD patients without CAD and studied the relationships between BNP levels and cardiac function and clinical variables. Plasma ANP levels significantly decreased after HD on Monday, Wednesday, and Friday, and predialysis plasma ANP levels on Monday were significantly greater than those on other days. Plasma BNP levels did not change after HD on Monday; however, they significantly decreased after HD on Wednesday and FRIDAY: Predialysis plasma BNP levels on Monday were greater than those on other days, and postdialysis plasma BNP levels on Monday were greater than predialysis plasma BNP levels on WEDNESDAY: Plasma BNP levels in HD patients with CAD were significantly greater than those in HD patients without CAD and significantly correlated with left ventricular (LV) ejection fraction (r = -0.69), end-diastolic volume index (r = 0.59), and end-systolic volume index (r = 0.84) determined by left ventriculography. Conversely, plasma BNP levels in HD patients without CAD significantly correlated with LV mass index (r = 0.54) determined by echocardiography and mean systolic blood pressure (r = 0.72) determined by 48 hour ambulatory blood pressure monitoring. These results suggest the following: (1) plasma BNP levels before and after HD in chronic HD patients directly correlate with the degree of body fluid retention, and the day of the week on which the sample is obtained should be considered for its evaluation; (2) plasma BNP levels reflect LV function in HD patients with CAD; and (3) plasma BNP levels reflect LV mass and blood pressure in HD patients without CAD. PMID- 11382690 TI - Effect of lipid-lowering dietary recommendations on the nutritional intake and lipid profiles of chronic peritoneal dialysis and hemodialysis patients. AB - Patients with end-stage renal failure are a high-risk group for atherosclerotic cardiovascular disease and commonly have dyslipidemia as a major factor. Dietary manipulation is the recommended first line of therapy for reducing lipid levels in people with normal renal function; however, complex dietary requirements of dialysis-treated patients with end-stage renal failure impose significant constraints. In this study, we evaluated the effect of trying to comply with established lipid-lowering recommendations superimposed on our normally prescribed dialysis diet over 14 weeks in stable subjects treated with either hemodialysis (HD) or chronic peritoneal dialysis (PD). Of 306 dialysis patients screened, 75 subjects were enrolled; 8 subjects did not complete the study. In the remainder, HD subjects (n = 41) decreased saturated fat intakes by 18% overall and cholesterol intakes by 16%. This was associated with a decrease in total cholesterol levels from 232 +/- 8 to 209 +/- 4 mg/dL (mean +/- SEM; P = 0.007) and low-density lipoprotein cholesterol levels from 147 +/- 4 to 131 +/- 4 mg/dL (P = 0.009). However, energy intakes decreased by almost 10%. There were no statistically significant changes in PD patients (n = 26). Only 24.4% of HD (10 of 41 patients) and 15.4% of PD patients (4 of 26 patients) normalized their lipid levels. Considerable problems were encountered in maintaining compliance with the modified dialysis diets. This study shows that if adhered to, properly constructed dialysis diets are close to optimal lipid-lowering recommendations. Further dietary manipulation is difficult, leads to little benefit in the majority, and is accompanied by added problems of adherence. We conclude that the vast majority of dyslipidemic patients with end-stage renal failure require pharmacological therapy. PMID- 11382691 TI - Dialysis patients' preferences regarding cardiopulmonary resuscitation and withdrawal of dialysis in Japan. AB - The aim of this study is to show the preferences of Japanese dialysis patients for receiving cardiopulmonary resuscitation (CPR) in their current health status, if they were severely demented, or if they had terminal cancer and to determine their desires about continuing dialysis if they were severely demented or had terminal cancer. A questionnaire survey including the three scenarios was administered to 450 dialysis patients in 15 hospitals in JAPAN: Three hundred ninety-eight patients completed the questionnaires for a response rate of 88%. The majority of responding patients were men and were undergoing hemodialysis. Only 5% of the patients had discussed their preferences regarding CPR with their physicians, and 29%, with their family members. Forty-two percent of the patients answered that they would want to receive CPR if they experienced cardiopulmonary arrest in their current health status, and 12% answered in the affirmative if they were seriously demented or had terminal cancer. Eighteen percent of the patients would want to continue dialysis if they were demented, and 45%, if they had terminal cancer. Statistical analysis showed that more patients who were working tended to want to continue dialysis if they had terminal cancer than those who were not (53% versus 37%; P < 0.014). Patients' age and preferences did not statistically correlate. Preferences of Japanese dialysis patients for CPR and dialysis vary according to differences in health status, and only a minority would want to receive CPR for cardiopulmonary arrest even in their current health status. PMID- 11382692 TI - Morbidity and cost implications of inadequate hemodialysis. AB - American hemodialysis patients have short lifespans, frequent hospitalizations, and aggregate Medicare inpatient expenditures of $4 billion/year. Dose of dialysis, as quantified by the parameter, Kt/V, corresponds strongly with survival and is estimated to be inadequate (Kt/V <1.2) in one fourth of patients. However, little is known about the morbidity and cost implications of inadequate dialysis. We sought to determine the independent relationship between dose of dialysis and (1) number of hospitalizations, (2) hospital days, and (3) Medicare inpatient reimbursements. We randomly selected 674 patients from all 22 hemodialysis units in northeast Ohio and examined hospitalizations, hospital days, and Medicare inpatient reimbursements for a 6-month interval following a 90 day quantification of dialysis dose. Every 0.1 decrease in Kt/V was independently associated with more hospitalizations (rate ratio, 1.11; 95% confidence interval [CI], 1.07 to 1.15), increased hospital days (rate ratio, 1.12; 95% CI, 1.03 to 1.22), and higher Medicare inpatient expenditures ($940; 95% CI, $450 to $1,440) after adjustment for patient age, sex, race, cause of renal failure, number of years on dialysis, and number of comorbid conditions. We estimate that increasing dialysis doses to a Kt/V of 1.2 for all patients nationally may decrease Medicare inpatient expenditures by $150 million annually. In conclusion, inadequate dialysis dose is independently associated with increased hospitalizations, hospital days, and Medicare inpatient expenditures. Improving dialysis adequacy may both improve patient morbidity and lessen health care costs. PMID- 11382693 TI - A prospective study of vascular access infections at seven outpatient hemodialysis centers. AB - Vascular access infections are a major cause of morbidity and mortality in hemodialysis patients, and the use of antimicrobials to treat such infections contributes to the emergence and spread of antimicrobial-resistant bacteria. To determine the incidence of and risk factors for vascular access infections, we studied hemodialysis patients at 7 outpatient dialysis centers (4 in Richmond, VA, and 3 in Baltimore, MD) during December 1997 to July 1998. Vascular access infections were defined as local signs (pus or redness) at the vascular access site or a positive blood culture with no known source other than the vascular access; and hospitalization or receipt of an intravenous (IV) antimicrobial. A total of 796 patients were followed for 4,134 patient-months. The vascular access infection rate was 3.5/100 patient-months, ie, patients had a 3.5% risk of infection each month. Independent risk factors were the specific dialysis unit where the patient was treated (relative hazard varying from 1.0 to 4.1 among the 7 centers), catheter access (relative hazard, 2.1 v implanted access), albumin level (relative hazard, 2.4 for lowest v highest quartile), urea reduction ratio (relative hazard, 2.2 for lowest v highest quartile), and hospitalizations during the previous 90 days (relative hazard, 4.9 for >/=6 v zero hospitalizations). These data confirm that vascular access infections are common in hemodialysis patients and that infection rates differ substantially among different centers. Catheter use should be minimized to reduce these infections. Additionally, the possibility that improved serum albumin and urea reduction ratio could reduce vascular access infections should be evaluated. PMID- 11382694 TI - Retrograde hemodialysis access flow during dialysis as a predictor of access pathology. AB - We have detected retrograde flow by Doppler ultrasound within the hemodialysis access of 10 patients during routine dialysis. Nine of these accesses were prosthetic grafts, and 1 was an autogenous fistula. All 9 of the grafts had access pathology demonstrated by angiography. The single patient with the fistula exhibiting reversed access flow had a poorly developed access with no focal stenoses on angiogram, but 18% recirculation by ultrasound dilution. All patients with retrograde flow had access flow rates below 650 mL/min. While 3 of these patients had substantial access recirculation (2 grafts, 12% and 40%; and 1 fistula, 18%) 2 of these patients had 3% recirculation (2 needle urea method), and 4 patients had 0% access recirculation (ultrasound dilution and 2 needle urea method). We report that retrograde access flow during dialysis may be a specific indicator of access dysfunction. These findings further suggest that retrograde access flow develops before access recirculation, indicating that this finding is more sensitive than recirculation for detecting access dysfunction. Further study is needed to determine the utility of this finding in access surveillance. PMID- 11382695 TI - A comparison of quantitative computed tomography and dual X-ray absorptiometry for evaluation of bone mineral density in patients on chronic hemodialysis. AB - Chronic renal failure leads to a reduction of bone mineral density (BMD). Therefore, noninvasive methods to evaluate BMD are also used regularly in this patient population. In this study, we compared the results of two widely used methods, quantitative computed tomography (QCT) of the lumbar spine and dual x ray absorptiometry (DXA) at different sites in 90 patients on chronic hemodialysis. Additionally, we also determined various clinical and biochemical data to assess their relationship to BMD at the different measurement sites. A total of 75% of our patients was found to have reduced BMD, and 25% had an average z-score below -2. Z-scores obtained by the different methods and at the different measurement sites within an individual patient varied considerably from completely normal to severely reduced values. Multivariate analyses using clinical and biochemical parameters showed lower values of BMD at all measurement sites after transplantation and marginally higher values after parathyroidectomy, which was seen only in DXA measurement at the femoral neck. We conclude from our study that determination of BMD in chronic hemodialysis patients yields highly variable results, depending on the technique used. As long as the exact clinical relevance of these results has not been determined, regular routine measurement of isolated sites of BMD cannot be advocated in this patient population. PMID- 11382696 TI - Impact of co-amoxiclav on polymorphonuclear granulocytes from chronic hemodialysis patients. AB - Phagocyte-dependent host defenses are frequently impaired in maintenance hemodialysis patients who show an increased susceptibility to infections. In these individuals, the course of infections can be more aggressive than in normal hosts, and the antibiotic of choice should have a high antimicrobial effect without impairing host defenses. Hence, in uremic patients, the antibiotic enhancement of phagocyte functions may be of potential clinical importance in the outcome of bacterial infections. Because we demonstrated previously that co amoxiclav had beneficial properties that result in enhancement of the microbicidal functions of human polymorphonuclear cells (PMNs) from healthy subjects, we investigated the influence of this combination on the activities of PMNs from chronic hemodialysis patients against Klebsiella pneumoniae, a human pathogen that can pose severe problems in patients whose immunity is impaired. PMNs from chronic dialysis patients showed a diminished in vitro phagocytic efficiency with a reduced phagocytosis and bactericidal activity towards intracellular K. pneumoniae compared with that seen in PMNs from healthy subjects. When co-amoxiclav was added to PMNs from chronic hemodialysis patients, it was able to restore the depressed primary functions of PMNs, resulting in a significant high increase in both phagocytosis or killing activity. A similar pattern was detected with PMNs collected from hemodialysis patients treated with co-amoxiclav. The results of the present study provide evidence that co-amoxiclav is able to induce stimulation of depressed phagocytic response of PMNs from patients on chronic hemodialysis, restoring their primary functions both in vitro and in vivo. PMID- 11382697 TI - Protein intake, control of serum phosphorus, and relatively low levels of parathyroid hormone in elderly hemodialysis patients. AB - In maintenance hemodialysis (MHD) patients, advanced age is a factor associated with relative hypoparathyroidism and an adynamic bone. However, the underlying mechanism remains elusive. The aim of the present study was to analyze whether the observed spontaneous decrease in protein intake in the elderly favors a better control of serum phosphorus (P) and parathyroid hormone (PTH) levels. A cross-sectional study including 207 MHD patients (mean age, 60 +/- 14; 59% males; time on dialysis, 61 +/- 55 months) dialyzed 3.5 to 4.5 hours 3 times a week using bicarbonate hemodialysis from 6 Spanish hemodialysis centers was performed. In 95 patients, the nutrient intake was recorded over a 5-day period, and average daily ingestion of nutrients was calculated using a computerized diet analysis system. One-way analysis of variance showed that serum phosphorus and intact PTH decreased with age. In addition, patients with serum phosphorus lower than 4 mg/dL as compared with those with serum phosphorus greater than 4 mg/dL were older (68 +/- 9 v 58 +/- 15 years, P < 0.001), had a lower protein (0.86 +/- 0.3 v 1.05 +/- 0.4 g/kg body weight, P < 0.01) and caloric intake (21.9 +/- 7.4 v 25.7 +/- 8.3 kcal/Kg body weight, P < 0.01), and had lower PTH levels (102 +/- 155 v 290 +/- 345 pg/mL, P < 0.001). An inverse and significant correlation was observed between age and protein intake (r = -0.48; P < 0.01), caloric intake (r = -0.37, P < 0.01), serum phosphorus concentration (r = -0.40; P < 0.01), and PTH levels (r = -0.26; P < 0.01). Additionally, a significant positive correlation was found between serum phosphorus and PTH levels (r = 0.40; P < 0.01). The results obtained in the present study suggest that a lower serum phosphorus level due to spontaneous reduction of protein intake might contribute to the relative low PTH levels observed in elderly hemodialysis patients. PMID- 11382698 TI - Calciphylaxis is associated with hyperphosphatemia and increased osteopontin expression by vascular smooth muscle cells. AB - Calciphylaxis or calcific uremic arteriolopathy (CUA) is a fatal disease in dialysis patients due to calcification of cutaneous blood vessels. The pathogenesis has been attributed to elevated parathyroid hormone (PTH). However, recent studies evaluating vascular calcification in nondialysis patients have found that the smooth muscle cells play an active role, including production of the bone matrix protein osteopontin. To examine the involvement of various clinical parameters and smooth muscle cells of CUA, we performed a case-control analysis comparing 10 CUA patients with our current dialysis patients. Available histologic sections were immunostained for osteopontin, markers of smooth muscle cells, endothelial cells, and macrophages. Compared with our current dialysis population, patients with CUA were more likely to be obese, white, and female (P < 0.02). Comparison of laboratory values found CUA patients with lower serum albumin, greater serum phosphorus, and greater calcium X phosphorus product (P < 0.01). In contrast, there was no difference in the concentration of PTH or calcium between the 2 groups. Immunostaining of calcified blood vessels showed that all calcified vessels stained positive for osteopontin, whereas all the noncalcifed vessels showed no osteopontin localization. Staining for smooth muscle alpha-actin decreased in the medial layer with calcification, with cells appearing to be sloughed off, leading to near occlusion of the vessel lumen. Our case-control study demonstrates that hyperphosphatemia and an elevated calcium X phosphorus product is associated with CUA. Histologic examination suggests that the calcification is associated with increased expression of osteopontin by smooth muscle cells. PMID- 11382699 TI - Hyaluronan and peritoneal ultrafiltration: a test of the "filter-cake" hypothesis. AB - Adding hyaluronan (HA) to the dialysis fluid seems to improve the efficiency of peritoneal dialysis (PD). This effect may be explained by the gradual formation of a HA "filter-cake" that decreases the tissue hydraulic conductivity. A filter cake (concentration hyperpolarization layer) can be formed when a large, slowly diffusible molecule, such as HA, is partly sieved by the pores of a membrane during the process of transmembrane ultrafiltration. The filter cake then forms at the "uphill" membrane-fluid interface, thereby increasing the resistance to fluid flow across the membrane. To test the filter-cake hypothesis, we investigated the effects of intraperitoneal (IP) HA on peritoneal small solute and water transport by administering HA either during the dwells or as incubations before PD dwells in rats. In the first set of experiments, HA, 0.01% (n = 7), 0.05% (n = 6), and 0.1% (n = 7) was given in 20 mL dialysis fluid (3.86% Dianeal). Control group was instilled with 20 mL of dialysis fluid. Evans Blue (EB) albumin was given as an intra-arterial (IA) bolus and (51)Cr-EDTA as an intravenous (IV) infusion. Plasma and dialysate were sampled up to 240 minutes to determine total peritoneal clearance (Cl), clearance from dialysate to plasma (Cl ->P) of (125)I-albumin (RISA), clearance from plasma to dialysate (Cl-->D) of EB albumin, and mass transfer area coefficients (MTAC or permeability-surface area products, PS) of (51)Cr-EDTA and glucose. Peritoneal ultrafiltration (UF) was determined from RISA dilution. In the second set of experiments, rats were first incubated with 4 mL of phosphate-buffered saline (PBS) or PBS containing 0.1% HA for 120 minutes. Rats were then dialyzed with HA-free PD fluid, and sampling of plasma and dialysate was performed for 60 minutes. For HA concentrations exceeding 0.01%, UF volumes increased with increasing doses of HA. Small solute MTACs and initial UF were unaffected when HA was either given during the dwell or as a preincubation. Compared with control, there was a significant decrease in RISA-Cl for 0.05% HA and 0.1% HA. Also, Cl-->P decreased significantly compared with control for 0.1% HA. In conclusion, the present data clearly demonstrate that small solute MTACs and the glucose-induced osmotic water transport occurring early in the dwell are not affected by HA. Only the back-filtration of fluid from peritoneum to plasma was affected. PMID- 11382700 TI - Impact of therapeutic plasma exchange on cyclosporine kinetics during membrane based lipid apheresis. AB - Cyclosporine is used widely as an immunosuppressant in transplant recipients and for various autoimmune diseases. In some cases, these patients require therapeutic plasma exchange (TPE). Cyclosporine is known to be highly bound to lipoproteins, and their removal by TPE would be expected to have an impact on drug dosing. We studied cyclosporine kinetics in a 54-year-old woman who is status post-cardiac transplant and has been receiving weekly TPE for familial hypercholesterolemia. We obtained serial measurements of cyclosporine, low density lipoproteins, and high-density lipoproteins at scheduled times related to the dosing of the medication on days that she received TPE versus a day she did not. We also measured cyclosporine, low-density lipoprotein, and high-density lipoprotein levels in the fixed volume (3.5 L) of the discarded plasma. Our results show a similar rate of decline of serum cyclosporine levels on TPE days as compared with a day without TPE. Net cyclosporine in the discarded plasma was found to be approximately 1 mg per treatment or less, a relatively insignificant amount when compared with the ingested daily dose of 150 to 250 mg twice a day. Despite substantial removal of lipoproteins, there is minimal impact of TPE on serum levels of cyclosporine, and dosage adjustment is not needed for patients undergoing this procedure. PMID- 11382701 TI - A patient with partial central diabetes insipidus: clarifying pathophysiology and designing treatment. AB - Studies were undertaken in a 32-year-old man who developed polyuria (4 L/d) a few days after a basal skull fracture; the condition persisted 1 year after the accident. The other major features were thirst, a plasma sodium of 143 mmol/L, 24 hour urine osmolality of 221 mOsm/kg H(2)O, and levels of vasopressin in plasma that were less than 0.5 pg/mL on 20 separate occasions. The 24-hour urine volume implied that the diagnosis was partial rather than complete central diabetes insipidus; however, several random urine samples had a much higher osmolality. An infusion of hypertonic saline led to the release of vasopressin and the excretion of concentrated urine. We propose that the basis for the lesion may be the transection of some, but not all, of the fibers connecting the osmostat and vasopressin release center. This partial transection could permit vasopressin to be secreted in response to a larger rise in plasma sodium concentration. This pathophysiologic analysis provided the basis for therapy to minimize the degree of polyuria. PMID- 11382702 TI - Successful use of thoracoscopic pericardiectomy in elderly patients with massive pericardial effusion caused by uremic pericarditis. AB - We report the use of thoracoscopic pericardiectomy to treat two elderly patients with massive pericardial effusion caused by uremic pericarditis. A 79-year-old man, admitted to our hospital complaining of dyspnea, was diagnosed with end stage renal failure and began maintenance hemodialysis. Although intensive hemodialysis was performed, the patient could not remain on hemodialysis because of severe hypotension during the procedure. Echocardiography revealed massive pericardial effusion and severe hypokinesis of the left ventricular wall. Pericardiocentesis was performed first, without success, followed by thoracoscopic pericardiectomy under general anesthesia. One month after the pericardiectomy, episodes of hypotension during hemodialysis improved, and dyspnea diminished. Echocardiography showed no pericardial effusion and improvement of left ventricular wall motion. Pericarditis is a fatal complication in patients with end-stage renal failure and patients on maintenance hemodialysis. The second patient received the same procedure with a similar improvement of clinical symptoms. These cases suggest that thoracoscopic pericardiectomy is a safe and effective treatment of pericardial effusion caused by uremic pericarditis in elderly patients on hemodialysis. PMID- 11382703 TI - Intraglomerular metastasis from pancreatic cancer. AB - Few case reports have shown the presence of metastatic tumor cells in renal glomeruli. We report one case with intraglomerular metastasis proved at renal biopsy. A 60-year-old man suffered from weight loss and fever of unknown origin. Urinalysis revealed proteinuria with cellular and granular casts. Because vasculitis was suspected, renal biopsy was performed. Presence of tumor cells occupying the glomerular capillary lumina was shown by means of light microscopy and electron microscopy. Laboratory findings revealed elevated leukocyte count (28.9 x 10(3)/mm(3)), serum granulocyte colony-stimulating factor (G-CSF) (77 pg/mL), and serum CA 19-9 (21,885 U/mL). The patient soon developed disseminated intravascular coagulation and died. Autopsy findings revealed pancreatic cancer showing positive staining for G-CSF and CA 19-9. Tumor cells in the glomerular capillary lumina showed positive staining for CA 19-9 and proliferating cell nuclear antigen (PCNA). These results suggest that the pancreatic tumor cells producing G-CSF were entrapped in the glomerular capillary lumina where they proliferated. This may have been the first step in renal metastasis. PMID- 11382704 TI - Transjugular biopsy in patients with combined renal and liver disease: making every organ count. PMID- 11382705 TI - East and West in the dialysis unit. PMID- 11382706 TI - Dysregulation of the calcium, phosphorus, parathyroid hormone, and vitamin D axis: what are the causes and risks? PMID- 11382707 TI - Doqi guidelines for nutrition in long-term peritoneal dialysis patients: a dissenting view. AB - The DOQI guidelines recommend a dietary protein intake of 1.2 to 1.3 g/kg/d in long-term peritoneal dialysis patients. A quantitative analysis of the relationship between this high protein intake and phosphorus intake shows that this daily load of protein is likely to lead to hyperphosphatemia in anuric patients with borderline dialysate clearance. Currently available phosphorus binders are not efficient enough to offset completely the adverse effects of a phosphorus-rich diet. An inverse correlation has been shown between serum phosphorus and outcome among dialysis patients, probably mediated by an increase in the Ca x P product. Evidence, theoretic or experimental, that such a high protein intake is by itself beneficial to dialysis patients is lacking. Because high protein intake may increase the daily load of phosphorus and predispose to hyperphosphatemia, it may lead to harmful consequences, and it should be modified. Using the general recommendation for normal subjects plus the usual excessive nitrogen loss during peritoneal dialysis, between 0.9 and 1 g/kg/d most likely would be an appropriate protein intake without exposing the patient to excessive phosphorus load. PMID- 11382708 TI - Dyslipidemia and hypertension: twin killers in renal vascular disease. PMID- 11382709 TI - The woman who loved well but not too wisely, or the vicissitudes of immunosuppression. PMID- 11382710 TI - Dialysis modality distribution in the United States. PMID- 11382711 TI - Re-evaluating our approach to calcium and phosphorus management in dialysis patients. PMID- 11382712 TI - Cuffed tunneled catheter as permanent hemodialysis access in Lucania. PMID- 11382715 TI - Pulse methylprednisolone, cyclosporine, and ace inhibitor therapy decreases proteinuria in two siblings with familial focal segmental glomerulosclerosis. AB - Familial focal segmental glomerulosclerosis (FSGS) is a heterogeneous renal disease characterized by proteinuria and an unremitting deterioration of renal excretory function. Previous studies showed corticosteroid unresponsiveness and a variable response to cyclophosphamide therapy. We hypothesized that treatment with pulse methylprednisolone therapy (PMT), alternate-day corticosteroids, and cyclosphosphamide or cyclosporine would decrease proteinuria in patients with familial FSGS. Two adolescent brothers, 13 and 16 years old, presented with nephrotic range proteinuria, but with normal renal excretory function. Both brothers had renal biopsies that showed FSGS with mesangial hypercellularity and tubular atrophy. Intravenous PMT, at doses of 1 g, was initiated per the Tune Mendoza protocol. Both patients received lisinopril therapy. One brother (case 1) was treated with PMT, alternate-day corticosteroids, and cyclophosphamide (total cumulative cyclophosphamide dose was 154.3 mg/kg). Urinary protein-to-urinary creatinine (UP/UC) ratios decreased from 6.79 to 3.79. Cyclosporine therapy decreased the UP/UC further from 2.48 to 0.76 at the end of PMT. The other brother (case 2), treated with PMT, alternate-day corticosteroids, and cyclosporine, experienced a decrease in UP/UC from 7.27 to 1.14. At the time of last evaluation, approximately 7 months after the last PMT dose, the UP/UC ratios were 0.27 (case 1) and 0.37 (case 2). PMT-attributable adverse effects were not severe. Both patients continued to receive oral cyclosporine and lisinopril after completion of PMT. PMT and cyclosporine therapy may reduce proteinuria, without decreasing renal excretory function, in some patients with familial FSGS. Further evaluation of cyclosporine therapy and PMT of patients with familial FSGS is warranted. PMID- 11382716 TI - Administration and clearance of amphotericin B during high-efficiency or high efficiency/high-flux dialysis. AB - Administration and clearance of amphotericin B infused during high-efficiency or high-efficiency/high-flux dialysis were studied in two end-stage renal disease patients requiring systemic antimycotic treatment for fungal peritonitis. Amphotericin B concentrations were measured in the arterial and venous dialysis ports as well as in the ultrafiltrate. Amphotericin B is poorly dialyzable while administered during hemodialysis sessions with high-efficiency (CA 210) or high efficiency/high-flux (CT 190 G) membranes. Amphotericin B infusion during hemodialysis was well tolerated and can be administered conveniently in an outpatient dialysis setting, avoiding prolonged hospitalization for parenteral antifungal therapy. PMID- 11382717 TI - Monitoring platelet function in glycoprotein IIB/IIIA inhibitor therapy. PMID- 11382718 TI - Modulation of C-reactive protein-mediated monocyte chemoattractant protein-1 induction in human endothelial cells by anti-atherosclerosis drugs. AB - BACKGROUND: C-reactive protein (CRP) induces adhesion molecule expression by endothelial cells. However, the effects of CRP on chemokine expression by endothelial cells are not known. METHODS AND RESULTS: We tested the effects of CRP on the production of the chemokines monocyte chemoattractant protein-1 (MCP 1) and RANTES in cultured human umbilical vein endothelial cells. The secretion of chemokines was assessed by ELISA. Incubation with 100 microgram/mL recombinant human CRP induced a 7-fold increase in MCP-1 but no change in RANTES secretion. We showed that the effect of CRP on MCP-1 was present even at 5 microgram/mL CRP, with stepwise increases as the CRP concentration was increased to 10, 50, and 100 microgram/mL. The effect of CRP on MCP-1 induction was not influenced by aspirin (at concentrations up to 1 mmol/L), but it was significantly inhibited by 5 micromol/L simvastatin. The peroxisome proliferator-activated receptor-alpha activators fenofibrate (100 micromol/L) and Wy-14649 (100 micromol/L) almost completely abolished the induction of MCP-1, but the peroxisome proliferator activated receptor-gamma activator ciglitazone had only a moderate effect. CONCLUSIONS: These results further strengthen the role of CRP in the pathogenesis of vascular inflammation and, likely, atherosclerosis and provide a crucial insight into a novel mechanism of action of anti-atherosclerosis drugs such as simvastatin and fenofibrate. PMID- 11382719 TI - Detection of coronary artery stenoses by contrast-enhanced, retrospectively electrocardiographically-gated, multislice spiral computed tomography. AB - BACKGROUND: Multislice spiral computed tomography (MSCT) with retrospectively ECG gated image reconstruction permits coronary artery visualization. We investigated the method's ability to identify high-grade coronary artery stenoses and occlusions. METHODS AND RESULTS: A total of 64 consecutive patients were studied by MSCT (4x1 mm cross-sections, 500-ms rotation, table feed 1.5 mm/rotation, intravenous contrast agent, retrospectively ECG-gated image reconstruction). All coronary arteries and side branches with a luminal diameter >/=2.0 mm were assessed concerning evaluability and the presence of high-grade stenoses (>70% diameter stenosis) or occlusions. Results were compared with quantitative coronary angiography. Of 256 coronary arteries (left main, left anterior descending, left circumflex and right coronary artery, including their respective side branches), 174 could be evaluated (68%). In 19 patients (30%), all arteries were evaluable. Artifacts caused by coronary motion were the most frequent reason for unevaluable arteries. Overall, 32 of 58 high-grade stenoses and occlusions were detected by MSCT (58%). In evaluable arteries, 32 of 35 lesions were detected, and the absence of stenosis was correctly identified in 117 of 139 arteries (sensitivity, 91%; specificity, 84%). If analysis was extended to all stenoses with >50% diameter reduction, sensitivity was 85% (40 of 47) and specificity was 76% (96 of 127). CONCLUSIONS: MSCT with retrospective ECG gating permits the detection of coronary artery stenoses with high accuracy if image quality is sufficient, but its clinical use may presently be limited due to degraded image quality in a substantial number of cases, mainly due to rapid coronary motion. PMID- 11382720 TI - Percutaneous in situ coronary venous arterialization: report of the first human catheter-based coronary artery bypass. AB - Diffuse coronary artery disease is frequently untreatable by coronary artery bypass or angioplasty. Many such "no-option" patients have been subjects for trials of angiogenesis using growth factor manipulation or laser injury. We think these novel revascularization strategies are limited by insufficient inflow to putative areas of new microvasculature and thus seek a more mechanical solution. We report the use of a catheter-based system for arterializing the adjacent anterior cardiac vein in a patient with chronic total occlusion of the left anterior descending coronary artery. A composite catheter system (phased-array ultrasound imaging system mounted on a catheter with extendable nitinol needle) was used to deliver an exchange-length intracoronary guidewire from the proximal left anterior descending coronary artery into the parallel anterior interventricular vein. Using standard angioplasty techniques, a fistula was then constructed from the proximal artery to the coronary vein using a self-expanding connector. The proximal vein was blocked with a novel self-expanding "blocker," thus precluding "steal" through the coronary sinus and forcing retroperfusion of the anterior wall. The procedure was completed without complication, and a follow up angiogram at 3 months confirmed continued patency of the arteriovenous connection. This patient, who had severe angina before the procedure, has been asymptomatic for 12 months. Percutaneous in situ venous arterialization may be an effective therapy for diffuse, "untreatable" coronary disease by supplying a robust inflow of arterialized blood via retroperfusion to severely ischemic myocardium. PMID- 11382721 TI - Plasma total cysteine as a risk factor for vascular disease: The European Concerted Action Project. AB - BACKGROUND: Elevated plasma total homocysteine (tHcy) is a risk factor for cardiovascular disease. Although cysteine is structurally similar and metabolically linked to tHcy, its relation to the risk of cardiovascular disease has received little attention. We studied the relation between plasma total cysteine (tCys) levels and the risk of vascular disease in the coronary, cerebral, and peripheral vessels. METHODS AND RESULTS: This case-control study included 750 patients with vascular disease and 800 age- and sex-matched control subjects recruited from 19 centers in 9 European countries. Conventional risk factors for cardiovascular disease were recorded. In addition, plasma levels of tCys, tHcy, folate, B(6), B(12), and creatinine were measured. Overall, a U shaped relationship was observed between tCys and risk of vascular disease. With the middle range of 250 to 275 micromol/L tCys used as the reference category, the adjusted risk of vascular disease at low (300 micromol/L) tCys levels was 1.6 (95% CI 1.1 to 2.3). Different shapes of the dose-response relationship were seen for the 3 vascular disease categories. The relation with peripheral vascular and cerebrovascular disease was U-shaped, whereas a weak positive relation was observed with coronary heart disease. CONCLUSIONS: Our data show a significant U shaped relationship between tCys and cardiovascular disease after adjustment for tHcy, creatinine, and other cardiovascular disease risk factors. PMID- 11382722 TI - Combination therapy with abciximab reduces angiographically evident thrombus in acute myocardial infarction: a TIMI 14 substudy. AB - BACKGROUND: Use of abciximab in combination with administration of thrombolytics has been shown to improve epicardial and microvascular coronary blood flow in acute myocardial infarction (AMI). As a potential mechanism, we hypothesized that combination therapy would reduce angiographically evident thrombus (AET) and would increase lumen diameter compared with thrombolytic monotherapy. METHODS AND RESULTS: Patients who received combination therapy in TIMI 14 (low-dose thrombolytic plus abciximab, n=732) were compared with patients who received thrombolytic monotherapy without abciximab in the TIMI 4, 10A, 10B, and 14 trials (n=1662). Thrombus burden was assessed 90 minutes after treatment, and quantitative angiography was performed in an angiographic core laboratory by investigators blinded to treatment assignment. The frequency of AET was reduced in patients who received abciximab combination therapy compared with thrombolytic monotherapy (26.6% versus 35.4%, P<0.001). Similar findings were observed when the analysis was restricted to patients with patent arteries (14.7% versus 20.8%, P=0.001). Residual percent diameter stenosis at 90 minutes was also improved in the abciximab therapy group both in patent arteries (64.6+/-16.6 versus 68.3+/ 14.8, P<0.001) and between patent and occluded arteries (69.3+/-19.5 versus 73.8+/-17.9, P<0.001). The absence of AET was associated with an increased frequency of >70% ST-segment resolution by 90 minutes (37.2%, 110/296 versus 18.9%, 54/286, P<0.001). CONCLUSIONS: Compared with thrombolytic monotherapy, combination therapy with abciximab reduces AET, which in turn is associated with reduced residual stenosis and improved ST-segment resolution in AMI. These data provide a pathophysiological link between platelet inhibition, reduced thrombus, and improvements in both epicardial and microvascular perfusion in AMI. PMID- 11382723 TI - Ability of recombinant factor VIIa to generate thrombin during inhibition of tissue factor in human subjects. AB - BACKGROUND: In view of the central role of the tissue factor-factor VIIa pathway in the initiation of blood coagulation, novel therapeutic strategies aimed at inhibiting this catalytic complex are currently being evaluated. A limitation of this new class of anticoagulants may be the lack of an appropriate strategy to reverse the effect if a bleeding event occurs. The aim of this study was to investigate the in vivo potential of recombinant factor VIIa (rVIIa) to induce thrombin generation in healthy subjects pretreated with recombinant nematode anticoagulant protein c2, a specific inhibitor of the tissue factor-factor VIIa complex, in a double-blind randomized crossover study. METHODS AND RESULTS: Administration of nematode anticoagulant protein c2 (3.5 microgram/kg) caused a prolongation of the prothrombin time from 13.7+/-0.6 to 16.9+/-1.2 seconds. The subsequent injection of rVIIa (90 microgram/kg) resulted in an immediate and complete correction of the prothrombin time and a marked generation of thrombin, reflected by increased levels of prothrombin activation fragment F1+2 and thrombin-antithrombin complexes from 0.75+/-0.64 to 3.29+/-6.3 nmol/L and from 2.4+/-0.6 to 10.7+/-3.9 microgram/mL, respectively. Factor X and IX activation peptides showed a 3.5-fold and a 3.8-fold increase, respectively, after the administration of rVIIa in the presence of nematode anticoagulant protein c2. CONCLUSIONS: During treatment with an inhibitor of the tissue factor-factor VIIa complex, the infusion of rVIIa resulted in thrombin generation. Our results indicate that rVIIa may be a good candidate as an antidote for inhibitors of tissue factor. PMID- 11382724 TI - Noninvasive quantification of coronary blood flow reserve in humans using myocardial contrast echocardiography. AB - BACKGROUND: We hypothesized that coronary blood flow (CBF) reserve could be quantified noninvasively in humans using myocardial contrast echocardiography (MCE). METHODS AND RESULTS: Eleven patients with normal epicardial coronary arteries (group I) and 19 with single-vessel coronary stenosis (group II) underwent quantitative coronary angiography, MCE, and CBF velocity measurements at rest and during intravenous adenosine infusion. In group I patients, MCE derived myocardial blood flow (MBF) velocity reserve (2.4+/-0.08) was similar to CBF velocity reserve using a Doppler flow wire (2.4+/-1.1). Patients with a single risk factor had a significantly higher MBF reserve (3.0+/-0.89) than those with >/=2 risk factors (1.7+/-0.22). In group II patients, significant differences were found in MBF velocity reserve in patients with mild (<50%), moderate (50% to 75%), or severe (>75%) stenoses (2.2+/-0.40, 1.6+/-0.65, and 0.55+/-0.19, respectively; P=0.005). A linear relation was found between flow velocity reserve determined using the 2 methods (r=0.76, P<0.001), and a curvilinear relation was noted between the percent coronary stenosis measured using quantitative coronary angiography and velocity reserve using both methods. CONCLUSIONS: CBF reserve can be measured in humans using MCE. This method may allow the noninvasive assessment of coronary stenosis severity and the detection of microvascular dysfunction. PMID- 11382725 TI - Prediction of mortality by exercise echocardiography: a strategy for combination with the duke treadmill score. AB - BACKGROUND: In studies generally involving short follow-up, exercise echocardiography has been shown to predict composite end points. We sought to study the prediction of mortality with this test and to devise a strategy for combination with standard exercise testing. METHODS AND RESULTS: Clinical, exercise testing, and echocardiographic data were collected in 5375 patients (aged 54+/-14 years, 3880 men) undergoing exercise echocardiography. The Duke treadmill score was derived from the results of treadmill exercise testing. Resting left ventricular (LV) function and the presence and severity of ischemia were interpreted by expert observers. Follow-up at 10.6 years (mean 5.5+/-1.9 years) was complete in 5211 patients (97%). The Duke score classified 59% of patients as low risk, 39% as intermediate risk, and 2% as high risk. Resting LV dysfunction was present in 1445 patients (27%), and the exercise echocardiogram was abnormal in 2525 patients (47%). Death occurred in 649 patients (12%). Over the first 6 years of follow-up, those with normal exercise echocardiograms had a mortality of 1% per year. Ischemia was an independent predictor of mortality. In sequential Cox models, the predictive power of clinical data was strengthened by adding the Duke score, resting LV function, and the results of exercise echocardiography. Exercise echocardiography was able to substratify patients with intermediate-risk Duke scores into groups with a yearly mortality of 2% to 7%. CONCLUSIONS: A normal exercise echocardiogram confers a low risk of death, and positive results are an independent predictor of death; ischemia is incremental to other data. This test may be particularly useful in patients with intermediate risk Duke treadmill scores. PMID- 11382726 TI - Point-of-care measured platelet inhibition correlates with a reduced risk of an adverse cardiac event after percutaneous coronary intervention: results of the GOLD (AU-Assessing Ultegra) multicenter study. AB - BACKGROUND: The optimal level of platelet inhibition with a glycoprotein (GP) IIb/IIIa antagonist necessary to minimize thrombotic complications in patients undergoing a percutaneous coronary intervention (PCI) is currently unknown. METHODS AND RESULTS: Five hundred patients undergoing a PCI with the planned use of a GP IIb/IIIa inhibitor had platelet inhibition measured at 10 minutes, 1 hour, 8 hours, and 24 hours after the initiation of therapy with the Ultegra Rapid Platelet Function Assay (Accumetrics). Major adverse cardiac events (MACES: composite of death, myocardial infarction, and urgent target vessel revascularization) were prospectively monitored, and the incidence correlated with the measured level of platelet function inhibition at all time points. One quarter of all patients did not achieve >/=95% inhibition 10 minutes after the bolus and experienced a significantly higher incidence of MACEs (14.4% versus 6.4%, P=0.006). Patients whose platelet function was <70% inhibited at 8 hours after the start of therapy had a MACE rate of 25% versus 8.1% for those >/=70% inhibited (P=0.009). By multivariate analysis, platelet function inhibition >/=95% at 10 minutes after the start of therapy was associated with a significant decrease in the incidence of a MACE (odds ratio 0.46, 95% CI 0.22 to 0.96, P=0.04). CONCLUSIONS: Substantial variability in the level of platelet function inhibition is achieved with GP IIb/IIIa antagonist therapy among patients undergoing PCI. The level of platelet function inhibition as measured by a point of-care assay is an independent predictor for the risk of MACEs after PCI. PMID- 11382727 TI - Different prognostic impact of 24-hour mean blood pressure and pulse pressure on stroke and coronary artery disease in essential hypertension. AB - BACKGROUND: We tested the hypothesis that the steady and pulsatile components of blood pressure (BP) exert a different influence on coronary artery disease and stroke in subjects with hypertension. METHODS AND RESULTS: We analyzed data on 2311 subjects with essential hypertension. All subjects (mean age 51 years, 47% women) underwent off-therapy 24-hour ambulatory BP monitoring. Over a follow-up period of up to 14 years (mean 4.7 years), there were 132 major cardiac events (1.20 per 100 person-years) and 105 cerebrovascular events (0.90 per 100 person years). After adjustment for age, sex, diabetes, serum cholesterol, and cigarette smoking (all P<0.01), for each 10 mm Hg increase in 24-hour pulse pressure (PP), there was an independent 35% increase in the risk of cardiac events (95% CI 17% to 55%). Twenty-four-hour mean BP was not a significant predictor of cardiac events after controlling for PP. After adjustment for age, sex, and diabetes (all P<0.05), for every 10 mm Hg increase in 24-hour mean BP, the risk of cerebrovascular events increased by 42% (95% CI 19% to 69%), and 24-hour PP did not yield significance after controlling for 24-hour mean BP. Twenty-four-hour PP was also an independent predictor of fatal cardiac events, and 24-hour mean BP was an independent predictor of fatal cerebrovascular events. CONCLUSIONS: In subjects with predominantly systolic and diastolic hypertension, ambulatory mean BP and PP exert a different predictive effect on the cardiac and cerebrovascular complications. Although PP is the dominant predictor of cardiac events, mean BP is the major independent predictor of cerebrovascular events. PMID- 11382728 TI - Permanent epicardial pacing in pediatric patients: seventeen years of experience and 1200 outpatient visits. AB - BACKGROUND: The purpose of this study was to evaluate the long-term outcome of all pediatric epicardial pacing leads. METHODS AND RESULTS: All epicardial leads and 1239 outpatient visits between January 1, 1983, and June 30, 2000, were retrospectively reviewed. Pacing and sensing thresholds were reviewed at implant, at 1 month, and at subsequent 6-month intervals. Lead failure was defined as the need for replacement or abandonment due to pacing or sensing problems, lead fracture, or phrenic/muscle stimulation. A total of 123 patients underwent 207 epicardial lead (60 atrial/147 ventricular, 40% steroid) implantations (median age at implant was 4.1 years [range 1 day to 21 years]). Congenital heart disease was present in 103 (84%) of the patients. Epicardial leads were followed for 29 months (range 1 to 207 months). The 1-, 2-, and 5-year lead survival was 96%, 90%, and 74%, respectively. Compared with conventional epicardial leads, both atrial and ventricular steroid leads had better stimulation thresholds 1 month after implantation; however, only ventricular steroid leads had improved chronic pacing thresholds (at 2 years: for steroid leads, 1.9 muJ [from 0.26 to 16 mu]; for nonsteroid leads, 4.7 muJ [from 0.6 to 25 muJ]; P<0.01). Ventricular sensing was significantly better in steroid leads 1 month after lead implantation (at 2 years: for steroid leads, 8 mV [from 4 to 31 mV]; for nonsteroid leads, 4 mV [from 0.7 to 10 mV]; P<0.01). Neither congenital heart disease, lead implantation with a concomitant cardiac operation, age or weight at implantation, nor the chamber paced was predictive of lead failure. CONCLUSIONS: Steroid epicardial leads demonstrated relatively stable acute and chronic pacing and sensing thresholds. In this evaluation of >200 epicardial leads, lead survival was good, with steroid-eluting leads demonstrating results similar to those found with historical conventional endocardial leads. PMID- 11382729 TI - Alleviation of myocardial ischemia after Kawasaki disease by heparin and exercise therapy. AB - BACKGROUND: Heparin promotes angiogenesis. We evaluated the effects of combined treatment with heparin and exercise on myocardial ischemia in the chronic stage of Kawasaki disease. METHODS AND RESULTS: This study was conducted in 7 patients (aged 6 to 19 years) who had a totally occluded coronary artery and stress induced myocardial ischemia in the collateral-dependent areas. Twice-daily exercise using a bicycle ergometer was performed with increments of 0.5 W/kg every 3 minutes up to maximal exertion for 10 days. Heparin, which immediately increased circulating hepatocyte growth factor, was given intravenously 10 minutes before each exercise period. Newly developed myocardial infarction, ventricular tachyarrhythmia, anginal attack, or hemorrhagic complication was not observed in any patient. Dipyridamole-loading single photon emission computed tomography documented improved myocardial perfusion in the collateral-dependent areas and a significant reduction in total defect scores in all patients after the completion of 20 sessions (P=0.01). In control patients who did not receive the heparin-exercise therapy, however, stress defect scores remained unchanged (n=1) or increased (n=2) during follow-up. Computerized quantitative coronary angiography provided evidence that the heparin-exercise therapy increased the diameter of the occluded artery to which collaterals terminated (P=0.001) but not that of the reference artery with which collaterals were not connected (P=0.96). CONCLUSIONS: The findings suggest that a series of heparin and exercise treatments over 10 days may have a dramatic effect on the alleviation of myocardial ischemia in collateral-dependent regions. This may be a safe, noninvasive revascularization therapy for patients with coronary artery occlusion in the chronic stage of Kawasaki disease. PMID- 11382730 TI - Simvastatin exerts both anti-inflammatory and cardioprotective effects in apolipoprotein E-deficient mice. AB - BACKGROUND: Simvastatin attenuates ischemia and reperfusion in normocholesterolemic animals by stabilizing endothelial nitric oxide synthase activity and inhibiting neutrophil-mediated injury. Because endothelial dysfunction is a detrimental effect of hypercholesterolemia, we examined whether short-term treatment with simvastatin could inhibit leukocyte-endothelium interaction and attenuate myocardial ischemia-reperfusion injury in apoE deficient (apoE(-/-)) mice fed a high-cholesterol diet. METHODS AND RESULTS: We studied leukocyte-endothelium interactions in apoE(-/-) mice fed a normal or a high-cholesterol diet after short-term (ie, 18 hours) simvastatin treatment. We also studied simvastatin treatment in myocardial ischemia-reperfusion injury by subjecting apoE(-/-) mice to 30 minutes of ischemia and 24 hours of reperfusion. ApoE(-/-) mice fed a high-cholesterol diet exhibited higher blood cholesterol levels, which were not affected by short-term simvastatin treatment. However, the increased leukocyte rolling and adherence that occurred in cholesterol-fed apoE( /-) mice (P<0.001 versus control diet) were significantly attenuated by simvastatin treatment (P<0.01 versus vehicle). Cholesterol-fed apoE(-/-) mice subjected to myocardial ischemia-reperfusion also experienced increased myocardial necrosis (P<0.01 versus control diet), which was significantly attenuated by simvastatin (P<0.01 versus vehicle). Simvastatin therapy also significantly increased vascular nitric oxide production in apoE(-/-) mice. CONCLUSIONS: Simvastatin attenuates leukocyte-endothelial cell interactions and ameliorates ischemic injury in hypercholesterolemic mice independently of lipid lowering actions. PMID- 11382731 TI - Acyl-CoA:cholesterol acyltransferase inhibition reduces atherosclerosis in apolipoprotein E-deficient mice. AB - BACKGROUND: Acyl-COA:cholesterol acyltransferase (ACAT) converts cholesterol to cholesteryl esters. The form of ACAT in macrophages, ACAT1, contributes to foam cell formation in the arterial wall and the development of atherosclerosis. Recent studies in a mouse model of atherosclerosis (the apolipoprotein E [apoE] deficient mouse), however, have suggested that complete deficiency of ACAT1 activity is not antiatherogenic, in part because of toxicity resulting from adverse effects on tissue cholesterol homeostasis. We have tested whether partial inhibition of ACAT1 and ACAT2 (expressed in liver and intestine) activities reduces atherosclerosis development in apoE-deficient mice and avoids toxicity. METHODS AND RESULTS: ApoE-deficient mice were maintained for 17 weeks on a Western-type diet without (control) or with the ACAT inhibitor F-1394 (effective against ACAT1 and ACAT2) at doses of either 300 (low) or 900 (high) mg/kg. Intimal lesion area at the aortic sinus in controls was 0.69+/-0.06 mm(2). F-1394 treatment significantly decreased lesional area by 39% (low) or 45% (high). F 1394 treatment also reduced lesional immunostaining for macrophages by 61% (low) or 83% (high). En face analysis showed that surface lipid staining in control aortas was 20.0+/-2.8%; F-1394 treatment reduced this by 46% (low) or 62% (high). There were no obvious signs of systemic or vessel wall toxicity associated with F 1394 treatment. CONCLUSIONS: Partial ACAT inhibition by F-1394 had antiatherogenic effects in apoE-deficient mice that were achieved without obvious toxicity. Partial ACAT inhibition may have therapeutic potential in the clinical treatment of atherosclerosis. PMID- 11382732 TI - T helper-cell phenotype regulates atherosclerosis in mice under conditions of mild hypercholesterolemia. AB - BACKGROUND: T cells are implicated in atherosclerosis, but little is known about the genetic control or molecular pathways, especially under conditions of mild hypercholesterolemia. METHODS AND RESULTS: BALB/c mice, making a CD4+ Th2 (IL-4+) cell response, express both MHC class II antigens (IA(d), IE(d)) and are atherosclerosis-resistant. C57Bl/6 mice produce a CD4+ Th1 (interferon [IFN]gamma+) response, express IA(b) but no IE, and are atherosclerosis-prone. To evaluate T helper-cell phenotype in fatty streak formation, wild-type C57Bl/6 mice (IA(b)+IE-) and transgenic mice, either AB(o), IA(b)-IE-; ABEalpha, IA IE(k)+; or BL:TG:Ealpha, IA(b)+IE(k)+, were fed a high-cholesterol diet for 16 weeks and evaluated histomorphometrically for aortic lesions. Lesion size in AB(o), ABEalpha, and BL:TG:Ealpha strains was decreased by 54%, 79%, and 82%, respectively, compared with wild-type, correlating with decreased Th1 and increased Th2 expression and suggesting that T helper-cell phenotype is important in fatty lesion development. Decreasing Th1 cells by antibodies (alpha-CD4) or cytokines (IL-4) also caused >/=80% reductions in lesion size. Immunohistology revealed IFN-gamma, but not IL-4, colocalized with activated macrophages. Confirming these findings in a different mouse strain, BALB/c Stat 6 knockout mice (Th2 cell-deficient) developed aortic lesions comparable to C57Bl/6 mice on the same diet. CONCLUSIONS: In mildly hypercholesterolemic C57Bl/6 mice, presence of IA(b) and absence of IE regulated CD4+ T helper-cell phenotype; fatty lesions were proportional to IFNgamma+ Th1 cells in both C57Bl/6 and BALB/c strains. IFN gamma may participate through macrophage activation, whereas IL-4 may act to limit Th1-cell response. PMID- 11382733 TI - Mitochondria as targets for nitric oxide-induced protection during simulated ischemia and reoxygenation in isolated neonatal cardiomyocytes. AB - BACKGROUND: As shown previously, exposure to NO donors initiates protective mechanisms in cardiomyocytes that persist after removal of the donor, a form of pharmacological preconditioning. Because NO also affects mitochondrial respiration, we studied the effect of NO on mitochondrial Ca(2+) uptake. METHODS AND RESULTS: Neonatal rat ventricular myocytes in primary culture were exposed to 1 hour of simulated ischemia and 1 hour of reoxygenation (sI/R). Pretreatment with the NO donor S-nitroso-N-acetyl-penicillamine (SNAP) (1 mmol/L for 90 minutes), followed by washing and incubation for 10 to 30 minutes, reduced sI/R induced cell death to 25.4% compared with control (propidium iodide exclusion assay, P<0.001). Short (10-second) exposures to SNAP reversibly suppressed mitochondrial respiration without a detectable change in mitochondrial potential. In contrast, treatment with SNAP for 90 minutes caused a modest but sustained mitochondrial depolarization, as judged by JC-1 fluorescence. SNAP pretreatment limited cellular Ca(2+) overload during ischemia (fura-2 ratio rose to 226+/-40% versus 516+/-170% of baseline, n=5, P<0.05) and prevented loss of cell membrane integrity during reoxygenation. SNAP pretreatment also significantly reduced the ability of mitochondria to accumulate Ca(2+) in the face of a similar cytosolic Ca(2+) load (peak rhod-2 fluorescence 133+/-4% versus 166+/-7% of baseline at similar fluo-3 levels, P=0.0004, n=52 and 25, respectively). CONCLUSIONS: Pretreatment with an NO donor induces a modest, sustained mitochondrial depolarization and protects cardiomyocytes from sI/R injury. The demonstrated reduction in mitochondrial Ca(2+) uptake possibly reduces cytosolic Ca(2+) overload, providing a likely mechanism for NO-induced protection. PMID- 11382734 TI - Detection of coronary artery stenosis with power Doppler imaging. AB - BACKGROUND: Power Doppler is a new imaging method for detecting microbubbles during myocardial contrast echocardiography (MCE) based on the registration of variance resulting from ultrasound-induced nonlinear bubble behavior. We tested the hypothesis that power Doppler imaging can be used to quantify coronary stenoses. METHODS AND RESULTS: Three left anterior descending (LAD) coronary stenoses of varying severity were created in each of 9 open-chest dogs. MCE was performed by continuous intravenous infusion of a nitrogen-filled bilayer shell microbubble, PB127, during triggered power Doppler imaging at incremental pulsing intervals. MCE and radiolabeled microsphere measurements were made at baseline and during each stenosis, with and without adenosine stress. Videointensities in the LAD and left circumflex (LCx) beds were plotted against pulsing interval and fit to a previously described exponential function modeling microbubble destruction and replenishment, which was used to derive parameters of bubble velocity (beta) and peak plateau videointensity (A). Contrast defects matching the location of radiolabeled microsphere hypoperfusion were clearly seen, without need for image processing. The product of beta and A was linearly related to LAD/LCx flow (r=0.90, P<0.0001) and inversely related to stenosis gradient (r= 0.70, P<0.0001). Endocardial/epicardial flow ratios were visualized and quantifiable. CONCLUSIONS: As with B-mode harmonics, a model of microbubble destruction/replenishment can be applied to power Doppler data as a means to detect a broad range of stenoses. Image clarity and the lack of attenuation or requirement for background subtraction are additional advantages of this imaging approach. Power Doppler MCE imaging holds promise for the detection of coronary artery disease. PMID- 11382735 TI - Left-to-right gradient of atrial frequencies during acute atrial fibrillation in the isolated sheep heart. AB - BACKGROUND: Recent studies demonstrated spatiotemporal organization in atrial fibrillation (AF). We hypothesized that waves emanating from sources in the left atrium (LA) undergo fragmentation, resulting in left-to-right frequency gradient. Our objective was to characterize impulse propagation across Bachmann's bundle (BB) and the inferoposterior pathway (IPP) during AF. METHODS AND RESULTS: In 13 Langendorff-perfused sheep hearts, AF was induced in the presence of acetylcholine (ACh). Fast Fourier transform of optical and bipolar electrode recordings was performed. Frequency-dependent changes in the left-to-right dominant frequency (DF) gradient were studied by perfusing D600 (2 micromol/L) and by increasing ACh concentration from 0.2 to 0.5 micromol/L. BB and IPP were subsequently ablated. At baseline, a left-to-right decrease in DFs occurred along BB and IPP, resulting in an LA-right atrium (RA) frequency gradient of 5.7+/-1.4 HZ: Left-to-right impulse propagation was present in 81+/-5% and 80+/-10% of cases along BB and IPP, respectively. D600 decreased the highest LA frequency from 19.7+/-4.4 to 16.2+/-3.9 Hz (P<0.01) and raised RA DF from 8.6+/-2.0 to 10.7+/-1.8 Hz (P<0.05). An increase in ACh concentration increased the LA-RA frequency gradient from 4.9+/-1.8 to 8.9+/-1.8 Hz (P<0.05). Ablation of BB and IPP decreased RA DF from 10.9+/-1.2 to 9.0+/-1.5 Hz (P<0.01) without affecting LA DF (16.8+/-1.5 versus 16.9+/-1.8 Hz, P=NS). CONCLUSIONS: Left-to-right impulse propagation and frequency-dependent changes in the LA-RA frequency gradient during AF strongly support the hypothesis that this arrhythmia is the result of high-frequency periodic sources in the LA, with fibrillatory conduction away from such sources. PMID- 11382736 TI - Challenges posed by adults with repaired congenital heart disease. PMID- 11382737 TI - Recommendations for the management of intracranial arteriovenous malformations: a statement for healthcare professionals from a special writing group of the Stroke Council, American Stroke Association. PMID- 11382738 TI - Clinical trials in cardiovascular medicine. PMID- 11382739 TI - Double-chambered right ventricle. PMID- 11382740 TI - Estrogen stimulates angiogenesis in normoperfused skeletal muscle in rabbits. PMID- 11382741 TI - Wide-complex tachycardia. PMID- 11382742 TI - Intravascular ultrasound guidance for stent implantation. PMID- 11382743 TI - P-wave duration and dispersion analysis: methodological considerations. PMID- 11382744 TI - Tremor-induced ECG artifact mimicking ventricular tachycardia. PMID- 11382746 TI - Real-time monitoring of the interactions of transforming growth factor-beta (TGF beta ) isoforms with latency-associated protein and the ectodomains of the TGF beta type II and III receptors reveals different kinetic models and stoichiometries of binding. AB - Mature transforming growth factor-beta (TGF-beta) is proteolytically derived from the C terminus of a precursor protein. Latency-associated protein (LAP), the N terminal remnant of the TGF-beta precursor, is able to bind and neutralize TGF beta. Mature TGF-beta exerts its activity by binding and complexing members of two subfamilies of receptors, the type I and II receptors. In addition to these signaling receptors, TGF-beta can also interact with an accessory receptor termed the type III receptor. Using a surface plasmon resonance-based biosensor (BIAcore), we determined the mechanisms of interaction of four binding proteins (LAP, the type II and III receptor ectodomains (EDs), and a type II receptor ED/Fc chimera) with three TGF-beta isoforms, and we quantified their related kinetic parameters. Using global fitting based on a numerical integration data analysis method, we demonstrated that LAP and the type II receptor/Fc chimera interacted with the TGF-beta isoforms with a 1:1 stoichiometry. In contrast, the type II ED interactions with TGF-beta were best fit by a kinetic model assuming the presence of two independent binding sites on the ligand molecule. We also showed that the type III ED bound two TGF-beta molecules. Further experiments revealed that LAP was able to block the interactions of TGF-beta with the two EDs, but that the two EDs did not compete or cooperate with each other. Together, these results strongly support the existence of a cell-surface complex consisting of one type III receptor, two TGF-beta molecules, and four type II receptors, prior to the recruitment of the type I receptor for signal transduction. Additionally, our results indicate that the apparent dissociation rate constants are more predictive of the neutralizing potency of these TGF-beta-binding proteins (LAP, the type II and III receptor EDs, and the type II receptor/Fc chimera) than the apparent equilibrium constants. PMID- 11382747 TI - Crystal structure of mannanase 26A from Pseudomonas cellulosa and analysis of residues involved in substrate binding. AB - The crystal structure of Pseudomonas cellulosa mannanase 26A has been solved by multiple isomorphous replacement and refined at 1.85 A resolution to an R-factor of 0.182 (R-free = 0.211). The enzyme comprises (beta/alpha)(8)-barrel architecture with two catalytic glutamates at the ends of beta-strands 4 and 7 in precisely the same location as the corresponding glutamates in other 4/7 superfamily glycoside hydrolase enzymes (clan GH-A glycoside hydrolases). The family 26 glycoside hydrolases are therefore members of clan GH-A. Functional analyses of mannanase 26A, informed by the crystal structure of the enzyme, provided important insights into the role of residues close to the catalytic glutamates. These data showed that Trp-360 played a critical role in binding substrate at the -1 subsite, whereas Tyr-285 was important to the function of the nucleophile catalyst. His-211 in mannanase 26A does not have the same function as the equivalent asparagine in the other GH-A enzymes. The data also suggest that Trp-217 and Trp-162 are important for the activity of mannanase 26A against mannooligosaccharides but are less important for activity against polysaccharides. PMID- 11382748 TI - Ionizing radiation-induced apoptosis in ataxia-telangiectasia fibroblasts. Roles of caspase-9 and cellular inhibitor of apoptosis protein-1. AB - Ionizing radiation (IR) has been shown to induce apoptosis to a greater extent in a fibroblast cell line AT5BIVA derived from an individual with ataxia telangiectasia (AT) than in control fibroblasts. However, the signaling pathway that underlies IR-induced apoptosis in AT cells has remained unknown. The mechanism of apoptosis in response to gamma-irradiation has now been examined in three AT fibroblast lines (AT3BIVA, AT4BIVA, and AT5BIVA) derived from different individuals with AT. The apoptotic indexes of these cell lines at 72 h after irradiation were 12, 31, and 35%, respectively, compared with a value of 2.3% for control fibroblasts. Immunoblot analysis and fluorometric assays revealed that the extents of IR-induced activation of caspase-3 and caspase-9 were markedly greater in AT4BIVA and AT5BIVA cells than in AT3BIVA and control cells. Furthermore, the basal abundance of the apoptotic inhibitor, a cellular inhibitor of apoptosis proteins (c-IAP-1), was markedly reduced in AT4BIVA and AT5BIVA cells compared with that in AT3BIVA and control cells. The overexpression of either caspase-9 mutant forms or recombinant c-IAP-1 in AT5BIVA cells inhibited the IR-induced activation of caspases-3 and 9 and reduced the apoptotic index of the irradiated cells. These results indicate that the extent of IR-induced apoptosis in different AT cell lines is inversely related to the abundance of c IAP-1 and directly related to the extent of activation of caspases-3 and 9. PMID- 11382749 TI - Rpb4, a non-essential subunit of core RNA polymerase II of Saccharomyces cerevisiae is important for activated transcription of a subset of genes. AB - A major role in the regulation of eukaryotic protein-coding genes is played by the gene-specific transcriptional regulators, which recruit the RNA polymerase II holoenzyme to the specific promoter. Several components of the mediator complex within the holoenzyme also have been shown to affect activation of different subsets of genes. Only recently has it been suggested that besides the largest subunit of RNA polymerase II, smaller subunits like Rpb3 and Rpb5 may have regulatory roles in expression of specific sets of genes. We report here, the role of Rpb4, a non-essential subunit of core RNA polymerase II, in activation of a subset of genes in Saccharomyces cerevisiae. We have shown below that whereas constitutive transcription is largely unaffected, activation from various promoters tested is severely compromised in the absence of RPB4. This activation defect can be rescued by the overexpression of cognate activators. We have localized the region of Rpb4 involved in activation to the C-terminal 24 amino acids. We have also shown here that transcriptional activation by artificial recruitment of the TATA-binding protein (TBP) to the promoter is also defective in the absence of RPB4. Surprisingly, the overexpression of RPB7 (the interacting partner of Rpb4) does not rescue the activation defect of all the promoters tested, although it rescues the activation defect of the heat shock element containing promoter and the temperature sensitivity associated with RPB4 deletion. Overall, our results indicate that Rpb4 and Rpb7 play independent roles in transcriptional regulation of genes. PMID- 11382750 TI - Identification of core alpha 1,3-fucosylated glycans and cloning of the requisite fucosyltransferase cDNA from Drosophila melanogaster. Potential basis of the neural anti-horseadish peroxidase epitope. AB - For many years, polyclonal antibodies raised against the plant glycoprotein horseradish peroxidase have been used to specifically stain the neural and male reproductive tissue of Drosophila melanogaster. This epitope is considered to be of carbohydrate origin, but no glycan structure from Drosophila has yet been isolated that could account for this cross-reactivity. Here we report that N glycan core alpha1,3-linked fucose is, as judged by preabsorption experiments, indispensable for recognition of Drosophila embryonic nervous system by anti horseradish peroxidase antibody. Further, we describe the identification by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry and high performance liquid chromatography of two Drosophila N-glycans that, as already detected in other insects, carry both alpha1,3- and alpha1,6-linked fucose residues on the proximal core GlcNAc. Moreover, we have isolated three cDNAs encoding alpha1,3-fucosyltransferase homologues from Drosophila. One of the cDNAs, when transformed into Pichia pastoris, was found to direct expression of core alpha1,3-fucosyltransferase activity. This recombinant enzyme preferred as substrate a biantennary core alpha1,6-fucosylated N-glycan carrying two non reducing N-acetylglucosamine residues (GnGnF6; Km 11 microm) over the same structure lacking a core fucose residue (GnGn; Km 46 microm). The Drosophila core alpha1,3-fucosyltransferase enzyme was also shown to be able to fucosylate N glycan structures of human transferrin in vitro, this modification correlating with the acquisition of binding to anti-horseradish peroxidase antibody. PMID- 11382751 TI - A role for the extracellular signal-regulated kinase and p38 mitogen-activated protein kinases in interleukin-1 beta-stimulated delayed signal tranducer and activator of transcription 3 activation, atrial natriuretic factor expression, and cardiac myocyte morphology. AB - We have demonstrated that two hypertrophic agents, interleukin-1 beta (IL-1 beta) and leukemic inhibitory factor (LIF), altered cardiac myocyte morphology with striking similarity and prompted us to investigate the common actions of these cytokines. We compared the phosphorylation/activation of signal tranducer and activator of transcription 3 (STAT3), extracellular signal-regulated kinase (ERK), p38(MAPK), and c-Jun N-terminal kinase mitogen-activated protein kinases (MAPKs). The phosphorylation of STAT3 by IL-1 beta was delayed (>60 min), whereas the response to LIF was rapid (<10 min) and transient. We confirmed that IL-1 beta potently stimulated all three MAPK subfamilies. In contrast, LIF promoted strong activation of ERKs, marginal activation of p38(MAPK), and no c-Jun N terminal kinase activation. To test the roles of ERKs and p38(MAPK), myocytes were pretreated with PD98059 and SB203580. Either inhibitor alone prevented STAT3 phosphorylation, implicating ERKs and p38(MAPK) in the delayed STAT3 response to IL-1 beta. The interplay of MAPKs and STAT3 phosphorylation in regulating IL-1 beta-stimulated hypertrophy was investigated by evaluating the effect of MAPK inhibitors on atrial natriuretic factor (ANF) expression and myocyte morphology. The specific inhibition of either ERK or p38(MAPK) attenuated the IL-1 beta- or LIF-stimulated ANF expression by up to 70%. Inhibition was not further increased in the presence of both inhibitors. Furthermore, although individual inhibition of ERK or p38(MAPK) did not affect morphology, co-treatment with both inhibitors abrogated the hypertrophic morphology stimulated by IL-1 beta but not by LIF. Taken together, our data indicate that the activation of ERK and p38(MAPK) is essential in regulating a delayed STAT3 phosphorylation as well as changes in ANF expression and morphology that follow IL-1 beta treatment. Thus, the role of MAPKs in the hypertrophic response can be dictated at least partly by the nature of the hypertrophic agent employed. PMID- 11382752 TI - Yol082p, a novel CVT protein involved in the selective targeting of aminopeptidase I to the yeast vacuole. AB - The yeast vacuolar enzyme aminopeptidase I (API) is synthesized in the cytoplasm as a precursor (pAPI). Upon its assembly into dodecamers, pAPI is wrapped by double-membrane saccular structures for its further transport within vesicles that fuse with the vacuolar membrane and release their content in the vacuolar lumen. Targeting of API to the vacuole occurs by two alternative transport routes, the cvt and the autophagy pathways, which although mechanistically similar specifically operate under vegetative growth or nitrogen starvation conditions, respectively. We have studied the role of Yol082p, a protein identified by its ability to interact with API, in the transport of its precursor to the vacuole. We show that Yol082p interacts with mature API, an interaction that is strengthened by the amino extension of the API protein. Yol082p is required for targeting of pAPI to the vacuole, both under growing and short term nitrogen starvation conditions. Absence of Yol082p does not impede the assembly of pAPI into dodecamers, but precludes the enclosure of pAPI within transport vesicles. Microscopy studies show that during vegetative growth Yol082p is distributed between a cytoplasmic pool and a variable number of 0.13--0.27-microm round, mobile structures, which are no longer observed under conditions of nitrogen starvation, and become larger in cells expressing the inactive Yol082 Delta C32p, or lacking Apg12p. In contrast to the autophagy mutants involved in API transport, a Delta yol082 strain does not lose viability under nitrogen starvation conditions, indicating normal function of the autophagy pathway. The data are consistent with a role of Yol082p in an early step of the API transport, after its assembly into dodecamers. Because Yol082p fulfills the functional requisites that define the CVT proteins, we propose to name it Cvt19. PMID- 11382753 TI - Involvement of ITF2 in the transcriptional regulation of melanogenic genes. AB - In response to agouti signal protein, melanocytes switch from producing eumelanin to pheomelanin concomitant with the down-regulation of melanogenic gene transcription. We previously reported that a ubiquitous basic helix-loop-helix transcription factor, known as ITF2, is up-regulated during this switch, and we now report that treatment of melanocytes with melanocyte-stimulating hormone down regulates expression of ITF2. To more fully characterize the involvement of ITF2 in regulating melanogenic gene transcription, ITF2 sense or antisense constructs were introduced into melan-a melanocytes. Gene and protein expression analyses and luciferase reporter assays using promoters from melanogenic genes showed that up-regulation of ITF2 suppressed melanogenic gene expression as well as the expression of Mitf, a melanocyte-specific transcription factor. In addition, stable ITF2 sense transfectants had significant reductions in pigmentation and a less dendritic phenotype compared with mock transfectants. In contrast, ITF2 antisense-transfected melanocytes were more pigmented and more dendritic. These results demonstrate that up-regulation of ITF2 during the pheomelanin switch is functionally significant and reveal that differential expression of a ubiquitous basic helix-loop-helix transcription factor can modulate expression of melanogenic genes and the differentiation of melanocytes. PMID- 11382754 TI - Purification, characterization, and cDNA cloning of lipoate-activating enzyme from bovine liver. AB - In mammals, lipoate-activating enzyme (LAE) catalyzes the activation of lipoate to lipoyl-nucleoside monophosphate. The lipoyl moiety is then transferred to the specific lysine residue of lipoate-dependent enzymes by the action of lipoyltransferase. We purified LAE from bovine liver mitochondria to apparent homogeneity. LAE activated lipoate with GTP at a 1000-fold higher rate than with ATP. The reaction absolutely required lipoate, GTP, and Mg(2+) ion, and the reaction product was lipoyl-GMP. LAE activated both (R)- and (S)-lipoate to the respective lipoyl-GMP, although a preference for (R)-lipoate was observed. Similarly, lipoyltransferase equally transferred both the (R)- and (S)-lipoyl moieties from the respectively activated lipoates to apoH-protein. Interestingly, however, only H-protein carrying (R)-lipoate was active in the glycine cleavage reaction. cDNA clones encoding a precursor LAE with a mitochondrial presequence were isolated. The predicted amino acid sequence of LAE is identical with that of xenobiotic-metabolizing/medium-chain fatty acid:CoA ligase-III, but an amino acid substitution due to a single nucleotide polymorphism was found. These results indicate that the medium-chain acyl-CoA synthetase in mitochondria has a novel function, the activation of lipoate with GTP. PMID- 11382756 TI - The selectivity filter of the voltage-gated sodium channel is involved in channel activation. AB - Amino acids located in the outer vestibule of the voltage-gated Na+ channel determine the permeation properties of the channel. Recently, residues lining the outer pore have also been implicated in channel gating. The domain (D) IV P-loop residue alanine 1529 forms a part of the putative selectivity filter of the adult rat skeletal muscle (mu1) Na+ channel. Here we report that replacement of alanine 1529 by aspartic acid enhances entry to an ultra-slow inactivated state. Ultra slow inactivation is characterized by recovery time constants on the order of approximately 100 s from prolonged depolarizations and by the fact that entry to this state can be reduced by binding to the pore of a mutant mu-conotoxin GIIIA, suggesting that ultra-slow inactivation may reflect a structural rearrangement of the outer vestibule. The voltage dependence of ultra-slow inactivation in DIV A1529D is U-shaped, with a local maximum near -60 mV, whereas activation is maximal only above -20 mV. Furthermore, a train of brief depolarizations produces more ultra-slow inactivation than a single maintained depolarization of the same duration. These data suggest that ultra-slow inactivation emanates from "partially activated" closed states and that the P-loop in DIV may undergo a conformational change during channel activation, which is accentuated by DIV A1529D. PMID- 11382755 TI - Molecular characterization of mammalian homologues of class C Vps proteins that interact with syntaxin-7. AB - Vesicle-mediated protein sorting plays an important role in segregation of intracellular molecules into distinct organelles. Extensive genetic studies using yeast have identified more than 40 vacuolar protein sorting (VPS) genes involved in vesicle transport to vacuoles. However, their mammalian counterparts are not fully elucidated. In this study, we identified two human homologues of yeast Class C VPS genes, human VPS11 (hVPS11) and human VPS18 (hVPS18). We also characterized the subcellular localization and interactions of the protein products not only from these genes but also from the other mammalian Class C VPS homologue genes, hVPS16 and rVPS33a. The protein products of hVPS11 (hVps11) and hVPS18 (hVps18) were ubiquitously expressed in peripheral tissues, suggesting that they have a fundamental role in cellular function. Indirect immunofluorescence microscopy revealed that the mammalian Class C Vps proteins are predominantly associated with late endosomes/lysosomes. Immunoprecipitation and gel filtration studies showed that the mammalian Class C Vps proteins constitute a large hetero-oligomeric complex that interacts with syntaxin-7. These results indicate that like their yeast counterparts, mammalian Class C Vps proteins mediate vesicle trafficking steps in the endosome/lysosome pathway. PMID- 11382757 TI - Simian virus 40 large tumor antigen modulates the Raf signaling pathway. AB - The large tumor antigen of simian virus 40 (SVLT) is a potent oncogene. Although inactivation of the p53 and pRb tumor suppressors has been causally linked to the transforming properties of SVLT, its exact mechanism of action remains undefined. Previous data indicated that Ras is activated in SVLT-expressing cells. In this report we show that SVLT also increases Raf kinase activity in both insect and mammalian cells, thus identifying the Raf kinase as an additional target of SVLT. Our results further show that SVLT was still able to activate Raf in cells where Ras levels had been drastically reduced through expression of an antisense construct, indicating that SVLT may activate Raf at least partly by a mechanism that is independent of its stimulatory effect on Ras. PMID- 11382758 TI - A screening for high copy suppressors of the sit4 hal3 synthetically lethal phenotype reveals a role for the yeast Nha1 antiporter in cell cycle regulation. AB - A screening for multicopy suppressors of the G(1)/S blockage of a conditional sit4 hal3 mutant yielded the NHA1 gene, encoding a Na(+),K(+)/H(+) antiporter, composed of a transmembrane domain and a large carboxyl-terminal tail, which has been related to cation detoxification processes. Expression of either the powerful Saccharomyces cerevisiae Ena1 Na(+)/H(+)-ATPase or the Schizosaccharomyces pombe Sod2 Na(+)/H(+) antiporter, although increasing tolerance to sodium, was unable to mimic the Nha1 function in the cell cycle. Mutation of the conserved Asp residues Asp(266)-Asp(267) selectively abolished Na(+) efflux without modifying K(+) efflux and did not affect the capacity of Nha1 to relieve the G(1) blockage. Mutagenesis analysis revealed that the region near the carboxyl-terminal end of Nha1 comprising residues 800-948 is dispensable for sodium detoxification but necessary for transport of K(+) cations. Therefore, this portion of the protein contains structural elements that selectively modulate Nha1 antiporter functions. This region is also required for Nha1 to function in the cell cycle. However, expression of the closely related Cnh1 antiporter from Candida albicans, which also contains a long carboxyl-terminal extension, although allowing efficient K(+) transport does not relieve cell cycle blockage. This indicates that although the determinants for Nha1-mediated regulation of potassium transport and the cell cycle map very closely in the protein, most probably the function of Nha1 on cell cycle is independent of its ability to extrude potassium cations. PMID- 11382759 TI - The proamphiregulin cytoplasmic domain is required for basolateral sorting, but is not essential for constitutive or stimulus-induced processing in polarized Madin-Darby canine kidney cells. AB - In this study, the role of the amphiregulin precursor (pro-AR) cytoplasmic domain in the basolateral sorting and cell-surface processing of pro-AR in polarized epithelial cells has been investigated using Madin-Darby canine kidney cells stably expressing various human pro-AR forms. Our results demonstrate that newly synthesized wild-type pro-AR (50 kDa) is delivered directly to the basolateral membrane domain with >95% efficiency, where it is sequentially cleaved within the ectodomain to release several soluble amphiregulin (AR) forms. Analyses of a pro AR cytoplasmic domain truncation mutant (ARTL27) and two pro-AR secretory mutants (ARsec184 and ARsec190) indicated that the pro-AR cytoplasmic domain is not required for efficient delivery to the plasma membrane, but does contain essential basolateral sorting information. We show that the pro-AR cytoplasmic domain truncation mutant (ARTL27) is not sorted in polarized Madin-Darby canine kidney cells, with approximately 65% of the newly synthesized protein delivered to the apical cell surface. Under base-line conditions, ARTL27 was preferentially cleaved from the basolateral surface with 4-fold greater efficiency compared with cleavage from the apical membrane domain. However, ARTL27 ectodomain cleavage could be stimulated equivalently from either membrane domain by a variety of different stimuli. The metalloprotease inhibitor BB-94 could inhibit both base line and stimulus-induced ectodomain cleavage of wild-type pro-AR and ARTL27. These results indicate that the pro-AR cytoplasmic domain is required for basolateral sorting, but is not essential for ectodomain processing. Preferential constitutive cleavage of ARTL27 from the basolateral cell surface also suggests that the metalloprotease activity involved in base-line and stimulus-induced ARTL27 ectodomain cleavage may be regulated differently in the apical and basolateral membrane domains of polarized epithelial cells. PMID- 11382761 TI - Apg2p functions in autophagosome formation on the perivacuolar structure. AB - Autophagy is a degradative process in which cytoplasmic components are non selectively sequestered by double-membrane structures, termed autophagosomes, and transported to the vacuole. We have identified and characterized a novel protein Apg2p essential for autophagy in yeast. Biochemical and fluorescence microscopic analyses indicate that Apg2p functions at the step of autophagosome formation. Apg2p localizes to some membranous structure distinct from any known organelle. Using fluorescent protein-tagged Apg2p, we showed that Apg2p localizes to a dot structure close to the vacuole, where Apg8p also exists, but not on autophagosomes unlike Apg8p. This punctate localization of Apg2p depends on the function of Apg1p kinase, phosphatidylinositol 3-kinase complex and Apg9p. Apg2p(G83E), encoded by an apg2-2 allele, shows a severely reduced activity of autophagy and a dispersed localization in the cytoplasm. Overexpression of the mutant Apg2p lessens the defect in autophagy. These results suggest that the dot structure is physiologically important. Apg2p and Apg8p are independently recruited to the structure but coordinately function there to form the autophagosome. PMID- 11382760 TI - Apg2 is a novel protein required for the cytoplasm to vacuole targeting, autophagy, and pexophagy pathways. AB - To survive starvation conditions, eukaryotes have developed an evolutionarily conserved process, termed autophagy, by which the vacuole/lysosome mediates the turnover and recycling of non-essential intracellular material for re-use in critical biosynthetic reactions. Morphological and biochemical studies in Saccharomyces cerevisiae have elucidated the basic steps and mechanisms of the autophagy pathway. Although it is a degradative process, autophagy shows substantial overlap with the biosynthetic cytoplasm to vacuole targeting (Cvt) pathway that delivers resident hydrolases to the vacuole. Recent molecular genetics analyses of mutants defective in autophagy and the Cvt pathway, apg, aut, and cvt, have begun to identify the protein machinery and provide a molecular resolution of the sequestration and import mechanism that are characteristic of these pathways. In this study, we have identified a novel protein, termed Apg2, required for both the Cvt and autophagy pathways as well as the specific degradation of peroxisomes. Apg2 is required for the formation and/or completion of cytosolic sequestering vesicles that are needed for vacuolar import through both the Cvt pathway and autophagy. Biochemical studies revealed that Apg2 is a peripheral membrane protein. Apg2 localizes to the previously identified perivacuolar compartment that contains Apg9, the only characterized integral membrane protein that is required for autophagosome/Cvt vesicle formation. PMID- 11382762 TI - Evidence for a distinct inhibitory factor in the regulation of p53 functional activity. AB - Under normal conditions, tumor suppressor protein p53 exists in the cell in its latent form and is unable to function as a transcription factor. The allosteric model of p53 regulation postulates that the extreme portion of p53 carboxyl terminus (aa 364-393) binds to the core domain of the protein, thereby abrogating specific DNA binding in that region. In this study we propose an alternative mechanism of p53 functional regulation, which involves a separate molecule acting in trans to inhibit p53 transcriptional activity. Through the use of chimeric proteins of p53, p63gamma and p73beta, we show that the extreme COOH-terminal domain of p53 exerts a powerful and specific inhibitory effect on the p73- and p63-driven expression of a reporter gene. Moreover, fusion of p53 extreme COOH terminus to a completely unrelated transcriptional activator Gal4-VP16 also results in significant inhibition of transactivation activity. Since p73, p63, or Gal4-VP16 cannot associate with any part of the p53 molecule, we conclude that p53(aa 364-393) represses transcriptional activity of chimeric proteins and p53 itself through the binding of external negative modulator(s) in that region and not by the allosteric mechanism of regulation. In accordance with the "distinct inhibitor" hypothesis, the activity of wild type p53 is substantially increased by overexpression of chimeric proteins bearing p53(aa 364-393), which might be due to the competitive removal of transcriptional inhibitor(s). Our findings provide the basis for the identification of such negative modulators of p53 transcriptional activity. PMID- 11382763 TI - Regulation of the L-type calcium channel by alpha 5beta 1 integrin requires signaling between focal adhesion proteins. AB - The L-type calcium channel is the major calcium influx pathway in vascular smooth muscle and is regulated by integrin ligands, suggesting an important link between extracellular matrix and vascular tone regulation in tissue injury and remodeling. We examined the role of integrin-linked tyrosine kinases and focal adhesion proteins in regulation of L-type calcium current in single vascular myocytes. Soluble tyrosine kinase inhibitors blocked the increase in current produced by alpha(5) integrin antibody or fibronectin, whereas tyrosine phosphatase inhibition enhanced the effect. Cell dialysis with an antibody to focal adhesion kinase or with FRNK, the C-terminal noncatalytic domain of focal adhesion kinase, produced moderate (24 or 18%, respectively) inhibition of basal current but much greater inhibition (63 or 68%, respectively) of integrin enhanced current. A c-Src antibody and peptide inhibitors of the Src homology-2 domain or a putative Src tyrosine phosphorylation site on the channel produced similar inhibition. Antibodies to the cytoskeletal proteins paxillin and vinculin, but not alpha-actinin, inhibited integrin-dependent current by 65-80%. Therefore, alpha(5)beta(1) integrin appears to regulate a tyrosine phosphorylation cascade involving Src and various focal adhesion proteins that control the function of the L-type calcium channel. This interaction may represent a novel mechanism for control of calcium influx in vascular smooth muscle and other cell types. PMID- 11382764 TI - Identification of Src phosphorylation sites in the catenin p120ctn. AB - p120-catenin (p120(ctn)) interacts with the cytoplasmic tail of cadherins and is thought to regulate cadherin clustering during formation of adherens junctions. Several observations suggest that p120 can both positively and negatively regulate cadherin adhesiveness depending on signals that so far remain unidentified. Although p120 tyrosine phosphorylation is a leading candidate, the role of this modification in normal and Src-transformed cells remains unknown. Here, as a first step toward pinpointing this role, we have employed two dimensional tryptic mapping to directly identify the major sites of Src-induced p120 phosphorylation. Eight sites were identified by direct mutation of candidate tyrosines to phenylalanine and elimination of the accompanying spots on the two dimensional maps. Identical sites were observed in vitro and in vivo, strongly suggesting that the physiologically important sites have been correctly identified. Changing all of these sites to phenylalanine resulted in a p120 mutant, p120-8F, that could not be efficiently phosphorylated by Src and failed to interact with SHP-1, a tyrosine phosphatase shown previously to interact selectively with tyrosine-phosphorylated p120 in cells stimulated with epidermal growth factor. Using selected tyrosine to phenylalanine p120 mutants as dominant negative reagents, it may now be possible to selectively block events postulated to be dependent on p120 tyrosine phosphorylation. PMID- 11382765 TI - Conformational change in the vinculin C-terminal depends on a critical histidine residue (His-906). AB - A phospholipid-controlled interaction between the N-terminal and C-terminal domains of vinculin is thought to be a major mechanism that regulates binding activities of the protein. To probe the mechanisms underlying these interactions we used chemical modification and site-directed mutagenesis directed at histidine residues. Diethylpyrocarbonate (DEPC) modification of the C-terminal, but not the N-terminal, domain greatly decreased affinity of the N-terminal-C-terminal binding, implicating histidine residues in the C-terminal. Mutation of either or both C-terminal histidines (at positions 906 and 1026), however, did not affect N C binding at neutral pH. The H906A mutation did prevent DEPC effects and also prevented the normal decrease in binding affinity for the N-terminal at lower pH. We found that the wild type C-terminal domain, but not the H906A mutant, underwent a conformational change at pH 6.5, reflected in an altered circular dichroism spectrum and apparent oligomerization. Phospholipid also induced conformational changes in the wild type C-terminal domain but not in the H906A mutant, even though the mutant protein did bind to the phospholipid. Finally, the sensitivity of the N-C interaction to phospholipid was much reduced by the H906A mutation. These results show that H906 plays a key role in the conformational dynamics of the C-terminal domain and thus the regulation of vinculin. PMID- 11382766 TI - Dynamic modulation of cytoskeletal proteins linking integrins to signaling complexes in spreading cells. Role of skelemin in initial integrin-induced spreading. AB - Recently we showed that signaling across beta3-integrin leads to activation of calpain and formation of integrin clusters that are involved in Rac activation. The subsequent activation of Rac and Rho leads to the formation of focal complexes and focal adhesions, respectively. The goal of the present study was to determine whether different proteins link the integrin to the cytoskeleton in the different complexes. We show that talin is present in focal adhesions but not in the calpain-induced clusters. alpha-Actinin colocalized with integrin at various sites, including the calpain-induced clusters. Skelemin, a protein shown recently to interact with beta1- and beta3-integrin in vitro, colocalized with integrin in calpain-induced clusters but was absent from focal adhesions. Cells transiently expressing skelemin C2 motifs, which contain the integrin binding site, failed to form integrin clusters or to spread on a substrate for beta1- and beta3 integrins. These results 1) suggest a dynamic reorganization of integrin complexes during cell spreading, 2) show that different cytoskeletal proteins link integrins in different complexes, and 3) demonstrate that skelemin is responsible for linking integrin to the calpain-induced clusters, and 4) show that the integrin-skelemin interaction is essential for transmission of signals leading to the initial steps of cell spreading. PMID- 11382767 TI - Purification and characterization of native conventional kinesin, HSET, and CENP E from mitotic hela cells. AB - We have developed a strategy for the purification of native microtubule motor proteins from mitotic HeLa cells and describe here the purification and characterization of human conventional kinesin and two human kinesin-related proteins, HSET and CENP-E. We found that the 120-kDa HeLa cell conventional kinesin is an active motor that induces microtubule gliding at approximately 30 microm/min at room temperature. This active form of HeLa cell kinesin does not contain light chains, although light chains were detected in other fractions. HSET, a member of the C-terminal kinesin subfamily, was also purified in native form for the first time, and the protein migrates as a single band at approximately 75 kDa. The purified HSET is an active motor that induces microtubule gliding at a rate of approximately 5 microm/min, and microtubules glide for an average of 3 microm before ceasing movement. Finally, we purified native CENP-E, a kinesin-related protein that has been implicated in chromosome congression during mitosis, and we found that this form of CENP-E does not induce microtubule gliding but is able to bind to microtubules. PMID- 11382768 TI - The p14ARF tumor suppressor protein facilitates nucleolar sequestration of hypoxia-inducible factor-1alpha (HIF-1alpha ) and inhibits HIF-1-mediated transcription. AB - Oncogenic alterations can influence tumor cell survival partly by affecting the activity of the hypoxia-inducible factor-1 (HIF-1) transcription factor. The alpha subunit of HIF-1 was found to be frequently overexpressed in advanced tumors, which was proposed to help the adaptation of tumor cells to hypoxia. Here we show that an important tumor suppressor protein, p14ARF (alternative reading frame product of the INK4A locus) can directly inhibit the transcriptional activity of HIF-1 by sequestering its alpha subunit into the nucleolus. The interaction requires neither p53 nor HDM2. This is one of the first reports that describe the interaction of p14ARF with a protein besides HDM2, which may define a p53-independent tumor suppressor activity for p14ARF. PMID- 11382769 TI - Claudin promotes activation of pro-matrix metalloproteinase-2 mediated by membrane-type matrix metalloproteinases. AB - Genes associated with regulation of membrane-type matrix metalloproteinase-1 (MT1 MMP)-mediated pro-MMP-2 processing were screened in 293T cells by a newly developed expression cloning method. One of the gene products, which promoted processing of pro-MMP-2 by MT1-MMP was claudin-5, a major component of endothelial tight junctions. Expression of claudin-5 not only replaced TIMP-2 in pro-MMP-2 activation by MT1-MMP but also promoted activation of pro-MMP-2 mediated by all MT-MMPs and MT1-MMP mutants lacking the transmembrane domain (DeltaMT1-MMP). A carboxyl-terminal deletion mutant of pro-MMP-2 (proDeltaMMP-2) was processed to an intermediate form by MT1-MMP in 293T cells and was further converted to an activated form by introduction of claudin-5. In contrast to the stimulatory effect of TIMP-2 on pro-MMP-2 activation by MT1-MMP, activation of pro-MMP-2 by DeltaMT1-MMP in the presence of claudin-5 and proDeltaMMP-2 processing by MT1-MMP were both inversely repressed by expression of exogenous TIMP-2. These results suggest that TIMP-2 is not involved in cluadin-5-induced pro-MMP-2 activation by MT-MMPs. Stimulation of MT-MMP-mediated pro-MMP-2 activation was also observed with other claudin family members, claudin-1, claudin-2, and claudin-3. Amino acid substitutions or deletions in ectodomain of claudin-1 abolished stimulatory effect. Direct interaction of claudin-1 with MT1 MMP and MMP-2 was demonstrated by immunoprecipitation analysis. MT1-MMP was co localized with claudin-1 not only at cell-cell borders, but also at other parts of the cells. TIMP-2 enhanced cell surface localization of MMP-2 mediated by MT1 MMP, and claudin-1 also stimulated it. These results suggest that claudin recruits all MT-MMPs and pro-MMP-2 on the cell surface to achieve elevated focal concentrations and, consequently, enhances activation of pro-MMP-2. PMID- 11382770 TI - Etk/Bmx tyrosine kinase activates Pak1 and regulates tumorigenicity of breast cancer cells. AB - Etk/Bmx, a member of the Tec family of nonreceptor protein-tyrosine kinases, is characterized by an N-terminal pleckstrin homology domain and has been shown to be a downstream effector of phosphatidylinositol 3-kinase. P21-activated kinase 1 (Pak1), another well characterized effector of phosphatidylinositol 3-kinase, has been implicated in the progression of breast cancer cells. In this study, we characterized the role of Etk in mammary development and tumorigenesis and explored the functional interactions between Etk and Pak1. We report that Etk expression is developmentally regulated in the mammary gland. Using transient transfection, coimmunoprecipitation and glutathione S-transferase-pull down assays, we showed that Etk directly associates with Pak1 via its N-terminal pleckstrin homology domain and also phosphorylates Pak1 on tyrosine residues. The expression of wild-type Etk in a non-invasive human breast cancer MCF-7 cells significantly increased proliferation and anchorage-independent growth of epithelial cancer cells. Conversely, expression of kinase-inactive mutant Etk-KQ suppressed the proliferation, anchorage-independent growth, and tumorigenicity of human breast cancer MDA-MB435 cells. These results indicate that Pak1 is a target of Etk and that Etk controls the proliferation as well as the anchorage independent and tumorigenic growth of mammary epithelial cancer cells. PMID- 11382771 TI - Residual ataxia telangiectasia mutated protein function in cells from ataxia telangiectasia patients, with 5762ins137 and 7271T-->G mutations, showing a less severe phenotype. AB - We have assessed several ataxia Telangiectasia mutated (ATM)-dependent functions in cells derived from ataxia telangiectasia patients, carrying either an ATM 5762ins137 splice site or a 7271T-->G missense mutation, with a less severe phenotype compared with the classical disorder. ATM kinase in vitro, from 5762ins137 cells, showed the same specific activity as ATM in normal cells, but the protein was present at low levels. In contrast, mutant ATM kinase activity in the 7271T-->G cells was only about 6% that of the activity in normal cells, although the level of mutant protein expressed was similar to normal cells. Phosphorylation of the DNA double strand break repair proteins Nbs1 and hMre11, following DNA damage, was observed in normal and 7271T-->G cells but was almost absent in both 5762ins137 and classical ataxia telangiectasia cells. The kinetics of p53 response was intermediate between normal and classical ataxia telangiectasia cells in both the 7271T-->G and 5762ins137 cells, but interestingly, c-Jun kinase activation following DNA damage was equally deficient in cell lines derived from all the ataxia telangiectasia patients. Our results indicate that levels of ATM kinase activity, but not induction of p53 or c-Jun kinase activity, in these cells correlate with the degree of neurological disorder in the patients. PMID- 11382772 TI - Glycogen synthase kinase 3beta regulates GATA4 in cardiac myocytes. AB - Inactivation of glycogen synthase kinase 3beta (GSK3beta) is critical for transcription of atrial natriuretic factor (ANF) by beta-adrenergic receptors in cardiac myocytes. We examined the mechanism by which GSK3beta regulates ANF transcription. Stimulation of beta-adrenergic receptors induced nuclear accumulation of GATA4, whereas beta-adrenergic ANF transcription was suppressed by dominant negative GATA4, suggesting that GATA4 plays an important role in beta adrenergic ANF transcription. Interestingly, GATA4-mediated transcription was markedly attenuated by GSK3beta. GSK3beta physically associates with GATA4 and phosphorylates GATA4 in vitro. Overexpression of GSK3beta suppressed both basal and beta-adrenergic increases in nuclear expression of GATA4, whereas inhibition of GSK3beta by LiCl caused nuclear accumulation of GATA4, suggesting that GSK3beta negatively regulates nuclear expression of GATA4. The nuclear exportin Crm1 reduced nuclear expression of GATA4, and the reduction was enhanced by GSK3beta but not by kinase-inactive GSK3beta. Leptomycin B, an inhibitor for Crm1, increased basal nuclear GATA4 and suppressed GSK3beta-induced decreases in nuclear GATA4. These results suggest that GSK3beta negatively regulates nuclear expression of GATA4 by stimulating Crm1-dependent nuclear export. Inhibition of GSK3beta by beta-adrenergic stimulation abrogates GSK3beta-induced nuclear export of GATA4, causing nuclear accumulation of GATA4, which may represent an important signaling mechanism mediating cardiac hypertrophy. PMID- 11382773 TI - Mechanisms of endothelin receptor subtype-specific targeting to distinct intracellular trafficking pathways. AB - We recently reported that the endothelin (ET) receptor subtypes ET(A) and ET(B) are targeted to distinct intracellular destinations upon agonist stimulation (Bremnes, T., Paasche, J. D., Mehlum, A., Sandberg, C., Bremnes, B., and Attramadal, H. (2000) J. Biol. Chem. 275, 17596-17604). The ET(A) receptor was shown to follow the recycling route of transferrin, whereas ET(B) is targeted to lysosomes for degradation. In the present study we have investigated the mechanisms of ET receptor subtype-specific targeting to distinct intracellular trafficking pathways. Truncation mutants of the ET(A) and ET(B) receptors with deletions of the cytoplasmic carboxyl-terminal tail distal to the palmitoylation site were found to mediate inositol phosphate accumulation and to internalize upon agonist stimulation, although internalization occurred at a slower rate as compared with the wild-type receptors. However, the truncated ET(A) receptor was no longer able to undergo recycling. Rather, both truncation mutants were recognized by beta-arrestin for recruitment to endocytosis and were sorted to lysosomes by a dynamin-dependent internalization pathway. Furthermore, studies of chimeric ET(A) and ET(B) receptors where the cytoplasmic tail of ET(A) was swapped with the corresponding domain of ET(B), and vice versa, revealed that the cytoplasmic tail of ET(B) is required for efficient lysosomal sorting and that signals for targeting to recycling reside in the cytoplasmic tail of the ET(A) receptor. PMID- 11382774 TI - Alpha-helix 2 in the amino-terminal mad homology 1 domain is responsible for specific DNA binding of Smad3. AB - Smads, signal transducers of the transforming growth factor-beta (TGF-beta) superfamily proteins, directly bind to DNA and regulate transcription of target genes. Smad3 binds to CAGA box, whereas Smad1 and Smad5 preferentially bind to GC rich sequences. The beta-hairpin loop in the amino-terminal Mad homology 1 (MH1) domain is the direct DNA-binding site of Smad3; however, the amino acid sequences of the beta-hairpin loop of Smad3 and Smad1/5 are identical, suggesting that other regions may be responsible for the differential DNA binding of Smad3 and Smad1/5. To identify regions other than the beta-hairpin loop responsible for specific DNA binding of Smad3, we generated chimeras containing various regions of Smad3 and Smad1. Luciferase assays using a TGF-beta-responsive reporter (CAGA)9-MLP-Luc and gel-mobility shift assays using 3xCAGA as a probe revealed that alpha-helix 2 (H2) in the amino-terminal part of the MH1 domain plays an important role in specific DNA binding and transcriptional activation of Smad3. Luciferase assays using natural TGF-beta-responsive reporters also revealed the functional importance of H2 in the Smad3 MH1 domain in direct DNA binding. Smad3 thus binds to DNA directly through the beta-hairpin loop, and H2 supports specific DNA binding of Smad3. PMID- 11382775 TI - Friend of GATA 2 physically interacts with chicken ovalbumin upstream promoter TF2 (COUP-TF2) and COUP-TF3 and represses COUP-TF2-dependent activation of the atrial natriuretic factor promoter. AB - Friend of GATA (FOG)-2 is a multi-zinc finger transcriptional corepressor protein that binds specifically to GATA4. Gene targeting studies have demonstrated that FOG-2 is required for normal cardiac morphogenesis, including the development of the coronary vasculature, left ventricular compact zone, and heart valves. To better understand the molecular mechanisms by which FOG-2 regulates these cardiac developmental programs, we screened a mouse day 11 embryo library using a yeast two-hybrid interaction trap with the fifth and sixth zinc fingers of FOG-2 as bait. Using this approach, we isolated clones encoding the orphan nuclear receptors chicken ovalbumin upstream promoter-transcription factor (COUP-TF) 2 and COUP-TF3. COUP-TF2-null embryos die during embryonic development with defective angiogenesis and cardiac defects, a pattern that partly resembles the FOG-2-null phenotype. The interaction between COUP-TF2 and FOG-2 in mammalian cells was confirmed by co-immunoprecipitation of these proteins from transfected COS-7 cells. The sites of binding interaction between COUP-TF2 and FOG-2 were mapped to zinc fingers 5 and 6 and fingers 7 and 8 of FOG-2 and to the carboxyl terminus of the COUP-TF proteins. Binding to COUP-TF2 was specific because FOG-2 did not interact with the ligand-binding domains of retinoid X receptor alpha, glucocorticoid receptor, and peroxisome proliferating antigen receptor gamma, which are related to the COUP-TF proteins. Full-length FOG-2 markedly enhanced transcriptional repression by GAL4-COUP-TF2(117-414), but not by a COUP-TF2 repression domain mutant. Moreover, FOG-2 repressed COUP-TF2dependent synergistic activation of the atrial natriuretic factor promoter by both GATA4 and the FOG-2 independent mutant GATA4-E215K. Taken together, these findings suggest that FOG-2 functions as a corepressor for both GATA and COUP-TF proteins. PMID- 11382776 TI - A novel endo-beta-galactosidase from Clostridium perfringens that liberates the disaccharide GlcNAcalpha 1-->Gal from glycans specifically expressed in the gastric gland mucous cell-type mucin. AB - We found that commercially available sialidases prepared from Clostridium perfringens ATCC10543 were contaminated with an endoglycosidase capable of releasing the disaccharide GlcNAcalpha1-->4Gal from glycans expressed in the gastric gland mucous cell-type mucin. We have isolated this enzyme in electrophoretically homogeneous form from the culture supernatant of this organism by ammonium sulfate precipitation followed by affinity chromatography using a Sephacryl S-200 HR column. The enzyme was specifically retained by and eluted from the column with methyl-alpha-Glc. By NMR spectroscopy, the structure of the disaccharide released from porcine gastric mucin by this enzyme was established to be GlcNAcalpha1-->4Gal. The specificity of this enzyme as an endo beta-galactosidase was established by analyzing the liberation of GlcNAcalpha1- >4Gal from GlcNAcalpha1-->4Galbeta1-->4GlcNAcbeta1-->6(GlcNAcalpha1--> 4Galbeta1- >3)GalNAc-ol by mass spectrometry. Because this novel endo-beta-galactosidase specifically releases the GlcNAcalpha1-->4Gal moiety from porcine gastric mucin, we propose to call this enzyme a GlcNAcalpha1-->4Gal-releasing endo-beta galactosidase (Endo-beta-Gal(GnGa)). Endo-beta-Gal(GnGa) was found to remove the GlcNAcalpha1-->4Gal epitope expressed in gastric adenocarcinoma AGS cells transfected with alpha1,4-N-acetylglucosaminyltransferase cDNA. Endo-beta Gal(GnGa) should become useful for studying the structure and function of glycoconjugates containing the terminal GlcNAcalpha1-->4Gal epitope. PMID- 11382777 TI - Irx4 forms an inhibitory complex with the vitamin D and retinoic X receptors to regulate cardiac chamber-specific slow MyHC3 expression. AB - The slow myosin heavy chain 3 gene (slow MyHC3) is restricted in its expression to the atrial chambers of the heart. Understanding its regulation provides a basis for determination of the mechanisms controlling chamber-specific gene expression in heart development. The observed chamber distribution results from repression of slow MyHC3 gene expression in the ventricles. A binding site, the vitamin D response element (VDRE), for a heterodimer of vitamin D receptor (VDR) and retinoic X receptor alpha (RXR alpha) within the slow MyHC3 promoter mediates chamber-specific expression of the gene. Irx4, an Iroquois family homeobox gene whose expression is restricted to the ventricular chambers at all stages of development, inhibits AMHC1, the chick homolog of quail slow MyHC3, gene expression within developing ventricles. Repression of the slow MyHC3 gene in ventricular cardiomyocytes by Irx4 requires the VDRE. Unlike VDR and RXR alpha, Irx4 does not bind directly to the VDRE. Instead two-hybrid and co immunoprecipitation assays show that Irx4 interacts with the RXR alpha component of the VDR/RXR alpha heterodimer and that the amino terminus of the Irx4 protein is required for its inhibitory action. These observations indicate that the mechanism of atrial chamber-specific expression requires the formation of an inhibitory protein complex composed of VDR, RXR alpha, and Irx4 that binds at the VDRE inhibiting slow MyHC3 expression in the ventricles. PMID- 11382778 TI - High affinity binding of receptor-associated protein to heparin and low density lipoprotein receptor-related protein requires similar basic amino acid sequence motifs. AB - The 39-kDa receptor-associated protein (RAP) is a specialized chaperone for members of the low density lipoprotein receptor gene family, which also binds heparin. Previous studies have identified a triplicate repeat sequence within RAP that appears to exhibit differential functions. Here we generated a series of truncated and site-directed RAP mutants in order to define the sites within RAP that are important for interacting with heparin and low density lipoprotein receptor-related protein (LRP). We found that high affinity binding of RAP to heparin is mediated by the carboxyl-terminal repeat of RAP, whereas both the carboxyl-terminal repeat and a combination of amino and central repeats exhibit high affinity binding to LRP. Several motifs were found to mediate the binding of RAP to heparin, and each contained a cluster of basic amino acids; among them, an intact R(282)VSR(285)SR(287)EK(289) motif is required for high affinity binding of RAP to heparin, whereas two other motifs, R(203)LR(205)R(206) and R(314)ISR(317)AR(319), also contribute to this interaction. We also found that intact motifs of both R(203)LR(205)R(206) and R(282)VSR(285)SR(287)EK(289) are required for high affinity binding of RAP to LRP, with the third motif, R(314)ISR(317)AR(319), contributing little to RAP-LRP interaction. We conclude that electrostatic interactions likely contribute significantly in the binding of RAP to both heparin and LRP and that high affinity interaction with both heparin and LRP appears to require mostly overlapping sequence motifs within RAP. PMID- 11382779 TI - Age-associated differences in cardiovascular inflammatory gene induction during endotoxic stress. AB - Upon physiological stress, families of stress response genes are activated as natural defense mechanisms. Here, we show that induction of specific inflammatory genes is significantly dysregulated and altered in the heart of aged (24--26 month-old) versus young (4-month-old) mice experimentally challenged with a bacterial endotoxin, lipopolysaccharide (LPS, 1.5 mg/kg of body mass). Whereas the LPS-mediated induction of cardiac mRNA for tumor necrosis factor alpha or inducible nitric-oxide synthase showed no age-associated differences, the induction of interleukin-1 beta (IL-1 beta) and intracellular adhesion molecule-1 was modestly extended with aging, and the induction of IL-6 was significantly prolonged with aging. This age-associated phenomenon occurred gradually from 4 to 17 months of age and became more evident after 23 months of age. The age associated augmentation of the cardiac IL-6 induction was also dramatic at the protein level. Immunohistochemically, the LPS-induced cardiac IL-6 was localized mainly in the microvascular walls. Aged but not young mice showed a high mortality rate during these experiments. These results demonstrate that endotoxin mediated induction of specific inflammatory genes in cardiovascular tissues is altered with aging, which may be causally related to the increased susceptibility of aged animals to endotoxic stress. PMID- 11382780 TI - TcSCA complements yeast mutants defective in Ca2+ pumps and encodes a Ca2+-ATPase that localizes to the endoplasmic reticulum of Trypanosoma cruzi. AB - Intracellular Ca(2+) in Trypanosoma cruzi is mainly located in an acidic compartment named the acidocalcisome, which among other pumps and exchangers possesses a plasma membrane-type Ca(2+)-ATPase. Evidence for an endoplasmic reticulum-located Ca(2+) uptake has been more elusive and based on indirect results. Here we report the cloning and sequencing of a gene encoding a sarcoplasmic-endoplasmic reticulum-type Ca(2+)-ATPase from T. cruzi. The protein (TcSCA) predicted from the nucleotide sequence of the gene has 1006 amino acids and a molecular mass of 109.7 kDa. Several sequence motifs found in sarcoplasmic endoplasmic reticulum-type Ca(2+)-ATPases were present in TcSCA. Expression of TcSCA in yeast mutants deficient in the Golgi and vacuolar Ca(2+) pumps (pmr1 pmc1 cnb 1) restored growth on EGTA. Membranes were isolated from the pmr1 pmc1 cnb1 mutant transformed with TcSCA, and it was found that the TcSCA polypeptide formed a Ca(2+)-dependent and hydroxylamine-sensitive (32)P-labeled phosphoprotein of 110 kDa in the presence of [gamma-(32)P]ATP. Cyclopiazonic acid, but not thapsigargin, blocked this phosphoprotein formation. Transgenic parasites expressing constructs of TcSCA with green fluorescent protein exhibited co-localization of TcSCA with the endoplasmic reticulum proteins BiP and calreticulin. An endoplasmic reticulum location was also found in amastigotes and trypomastigotes using a polyclonal antibody against a COOH-terminal region of the protein. The ability of TcSCA to restore growth of mutant pmr1 pmc1 cnb 1 on medium containing Mn(2+) suggests that TcSCA may also regulate Mn(2+) homeostasis by pumping Mn(2+) into the endoplasmic reticulum of T. cruzi. PMID- 11382781 TI - Identification and adipocyte differentiation-dependent expression of the unique disialic acid residue in an adipose tissue-specific glycoprotein, adipo Q. AB - Recently, we have shown that alpha 2,8-linked disialic acid (diSia) residue occurs in glycoproteins more frequently than ever recognized (Sato, C., Fukuoka, H., Ohta, K., Matsuda, T., Koshino, R., Kobayashi K., Troy, F. A., II, and Kitajima, K. (2000) J. Biol. Chem. 275, 15422--15431). In the course of identification of the diSia-containing glycoproteins in mammals, the 30-kDa glycoprotein was found in bovine serum. The 30-kDa glycoprotein was shown to be the bovine adipo Q, an adipocyte-specific protein, based on the partial amino acid sequences and the immuno-cross-reactivity with the recombinant mouse adipo Q. The bovine adipo Q was shown to have no N-linked but O-linked glycan(s) containing the diSia epitope, Neu5Ac alpha 2-->8Neu5Ac alpha 2-->3Gal. Furthermore, the diSia epitope was also found in the mouse adipo Q in serum as well as in the 3T3-L1 cells that are fully differentiated into adipocytes. Notably, among the known alpha 2,8-sialyltransferases, only the alpha 2,8 sialyltransferase III mRNA was detected in the 3T3-L1 cells at any stages of differentiation, and the recombinant alpha 2,8-sialyltransferase III could sialylate the purified bovine adipo Q. Thus, this study clearly provides the new findings that adipo Q is the diSia-containing glycoprotein and a physiological substrate of alpha 2,8-sialyltransferase III, whose substrates have not been identified so far. PMID- 11382782 TI - Calpain cleavage promotes talin binding to the beta 3 integrin cytoplasmic domain. AB - Talin links integrin beta cytoplasmic domains to the actin cytoskeleton and is involved in the clustering and activation of these receptors. To understand how talin recognizes integrin beta cytoplasmic domains, we configured surface plasmon resonance methodology to measure the interaction of talin with the beta3 integrin cytoplasmic domain. Here we report that the N-terminal approximately 47-kDa talin head domain (talin-H) has a 6-fold higher binding affinity than intact talin for the beta3 tail. The affinity difference is mainly due to a difference in k(on). Calpain cleavage of intact talin released talin-H and resulted in a 16-fold increase in apparent K(a) and a 100-fold increase in apparent k(on). The increase in talin binding after cleavage was greater than predicted for stoichiometric liberation of free talin-H. This additional increase in binding was due to cooperative binding of talin-H and talin rod domain to the beta3 tail. Talin resembles ERM (ezrin, radixin, moesin) proteins in possessing an N-terminal FERM (band four-point-one, ezrin, radixin, moesin) domain. These data show that the talin FERM domain, like that in the ERM proteins, is masked in the intact molecule. Furthermore, they suggest that talin cleavage by calpain may contribute to the effects of the protease on the clustering and activation of integrins. PMID- 11382783 TI - Interaction of two structurally distinct sequence types with the clathrin terminal domain beta-propeller. AB - The amino-terminal domain of the clathrin heavy chain, which folds into a seven bladed beta-propeller, binds directly to several endocytic proteins via short sequences based on the consensus residues LLDLD. In addition to a single LLDLD based, type I clathrin-binding sequence, both amphiphysin and epsin contain a second, distinct sequence that is also capable of binding to clathrin directly. Here, we analyzed these sequences, which we term type II sequences, and show that the (257)LMDLA sequence in rat epsin 1 appears to be a weak clathrin-binding variant of the sequence (417)PWDLW originally found in human amphiphysin II. The structural features of the type II sequence required for association with clathrin are distinct from the LLDLD-based sequence. In the central segment of amphiphysin, the type I and type II sequences cooperate to effect optimal clathrin binding and the formation of sedimentable assemblies. Together, the data provide evidence for two interaction surfaces upon certain endocytic accessory proteins that could cooperate with other coat components to enhance clathrin bud formation at the cell surface. PMID- 11382784 TI - Association of tyrosine phosphatase SHP-2 with F-actin at low cell densities. AB - SHP-2 is an intracellular SH2 domain-containing protein-tyrosine phosphatase with an essential role in cell signaling. Here we demonstrate that localization of SHP 2 is regulated by cell density in a cell adhesion-dependent manner. When cells were plated at low densities, SHP-2 was distributed in Triton X-100-insoluble fractions, whereas it was totally soluble when cells were plated at high densities or when low density cells approached confluency. In all cases, the total protein level of SHP-2 was not changed. Fluorescent cell staining revealed that SHP-2 was co-localized with actin stress fibers to the cell peripheral at low cell densities but was diffused in the entire cytoplasm at high cell densities. Transient transfection of cells with truncated forms of SHP-2 demonstrated that the catalytic domain of the enzyme was responsible for the density-regulated distribution of SHP-2, but the catalytic activity was not required. An in vitro co-sedimentation study demonstrated direct binding of full length and SH2 domain-truncated forms of SHP-2 to F-actin. The data indicate that SHP-2 is regulated by cell density and that it may have a role in assembling and disassembling of the actin network. PMID- 11382785 TI - Skeletal muscle satellite cell characteristics in young and older men and women after heavy resistance strength training. AB - Skeletal muscle satellite cell proportions and morphology were assessed in healthy, sedentary young and older men and women in response to heavy resistance strength training (HRST). Fourteen young (20-30 years) men (n = 7) and women (n = 7) and 15 older (65-75 years) men (n = 8) and women (n = 7) completed 9 weeks of unilateral knee extension exercise training 3 days per week. Muscle biopsies were obtained from each vastus lateralis before and after training, with the nondominant leg serving as an untrained control. All four groups demonstrated a significant increase in satellite cell proportion in response to HRST (2.3 +/- 0.4% vs 3.1 +/- 0.4% for all subjects combined, before and after training, respectively; p < .05), with older women demonstrating the greatest increase (p < .05). Morphology data indicated a significant increase in the proportion of active satellite cells in after-training muscle samples compared with before training samples and with control leg samples (31% vs 6% and 7%, respectively; p < .05). The present results indicate that the proportion of satellite cells is increased after HRST in young and older men and women, with an exaggerated response in older women. Furthermore, the proportion of satellite cells that appear morphologically active is increased as a result of HRST. PMID- 11382786 TI - Differential influence of physical activity on lumbar spine and femoral neck bone mineral density in the elderly population. AB - This study investigates the relationship between lifetime physical activity and bone mineral density (BMD) at various sites in 129 healthy men and women aged 72.1 +/- 6.5 years. BMD was measured by dual energy x-ray absorptiometry, and physical activity was assessed by using the QUANTAP system (Quantification de l'Activite Physique), a standardized and structured computer-assisted interview tool designed to assess lifetime physical activity. Linear regression models controlling for age, gender, height, body mass, lean mass, and smoking habits were performed. Higher levels of sporting activity during youth were associated with greater lumbar spine BMD ( p < .001). Similarly, femoral neck BMD was greatest in subjects who reported regularly taking part in sports over the previous 20 years ( p <. 05) and during their whole lifetime ( p < 0.05). Sporting activity at the time of bone mass development increases subsequent lumbar spine BMD, and more recent sporting activity contributes to the preservation of femoral neck BMD. These results suggest that physical activity has a differential influence on BMD at different sites and at different ages, possibly related to the processes of bone construction and bone aging taking place at the time. PMID- 11382787 TI - Motor unit properties of nerve-intact extensor digitorum longus muscle grafts in young and old rats. AB - Impaired reinnervation has been implicated as the cause of the threefold disparity in the recovery of maximum force (P0) of standard muscle grafts in old compared with young rats. The specific, null hypothesis of this study is that compared with age-matched control extensor digitorum longus (EDL) muscles, nerve intact EDL muscle grafts in young and old rats show no evidence of an age-related impairment in reinnervation. Nerve-intact grafts were performed in 3-month-old and 23-month-old rats and were evaluated 60 days postoperatively. Compared with age-matched control EDL muscles, nerve-intact grafts in young and old rats showed no difference in muscle mass or motor unit numbers. The mean motor unit P0 for nerve-intact graft muscles in both age groups was significantly lower than that of age-matched control muscles. These data support our hypothesis that if axons are allowed to regenerate in an endoneurial environment, there is no evidence of an age-related impairment in muscle reinnervation. PMID- 11382788 TI - Aging lowers steady-state antioxidant enzyme and stress protein expression in primary hepatocytes. AB - It has been reported that the isolation and culture of primary hepatocytes can compromise cellular ability to constituitively express antioxidant enzyme (AE) genes, making it difficult to study their regulation ex vivo. In the present study, the steady-state expression of manganese-containing superoxide dismutase, copper- and zinc-containing superoxide dismutase, catalase, and glutathione peroxidase was assessed in primary hepatocytes isolated from young and senescent rats and cultured in MATRIGEL: There was no change in steady-state superoxide dismutase protein or activity levels in cells collected from young animals and cultured for 7 days. Catalase expression was initially increased, and then it declined 30%. In contrast, superoxide dismutase expression declined 60% and catalase expression declined 50% in cells from senescent animals. Constitutive and inducible 70-kDa heat shock protein expression increased coincident with declining AE levels in the young cells but not senescent cells. For both age groups, electron micrographs showed rounded hepatocytes with abundant rough endoplasmic reticulum, mitochondria, and peroxisomes. Hepatocytes were organized into clusters of 6-12 cells surrounding a large central lumen devoid of microvilli. Each cluster also contained smaller microvilli-lined lumens between adjacent hepatocytes that resembled canniculi. The plasma membranes of these lumens were sealed from the extracellular space by junctional complexes. Gap junctions in the plasma membrane suggest that hepatocytes were capable of intercellular communication. We conclude that the Matrigel system can be used to study AE regulation in primary hepatocytes from young and senescent animals, provided that experiments can be conducted within a time frame of 5-7 days in culture. These data also support the hypothesis that aging compromises hepatocellular ability to maintain AE status and upregulate stress protein expression. PMID- 11382789 TI - Unexpected effects of a heterozygous dnmt1 null mutation on age-dependent DNA hypomethylation and autoimmunity. AB - DNA methylation modifies gene expression. Methylation patterns are established during ontogeny, but they change with aging, usually with a net decrease in methylation. The significance of this change in T cells is unknown, but it could contribute to autoimmunity, senescence, or both. We examined the effects of a null mutation in DNA methyltransferase 1 (Dnmt1), a gene maintaining DNA methylation patterns, on immune aging. Whereas aged control mice developed hypomethylated DNA, autoimmunity, and signs of immune senescence as predicted, the knockout mice surprisingly increased DNA methylation and developed signs of autoimmunity and senescence more slowly. To identify potential mechanisms, we compared transcripts of DNA methyltransferase and methylcytosine binding protein family members in control and knockout mice. MeCP2, a methylcytosine binding protein involved in gene suppression and chromatin inactivation, was the only transcript differentially expressed between old knockout mice and controls, and thus it is a candidate for a gene product mediating these effects. PMID- 11382790 TI - Issues and dilemmas in the prevention and treatment of pressure ulcers: a review. AB - Considerable dogma and rhetoric, rather than evidence-based results, have accompanied recommendations for the prevention and treatment of pressure ulcers. Therapy for pressure ulcers is generally empiric, based on anecdotal experience, or borrowed from the treatment of patients with acute wounds. The treatment of pressure ulcers is problematic because of multiple comorbidities of patients, the chronic duration of pressure ulcers, and often by the physician's relative unfamiliarity with treatment options. Issues and dilemmas in the prevention of pressure ulcers center around risk assessment, means of pressure relief, and nutritional support. Similar issues in the treatment of pressure ulcers include implementing pressure relief, nutritional support, local wound care, the best method of debridement, diagnosing infection, the use of topical growth factors, and surgical treatment. The accumulating data for the prevention and management of pressure ulcers permits an outline of clinical strategies. PMID- 11382791 TI - Coping with population aging in the old continent--the need for european academic geriatrics. PMID- 11382792 TI - Bioelectrical impedance analysis estimation of water compartments in elderly diseased patients: the source study. AB - BACKGROUND: This study validates, in geriatric patients, bioelectrical impedance analysis (BIA) equations that had been derived to estimate total body water (TBW) and extracellular water (ECW) in healthy elderly subjects. METHODS: We performed a multicentric trial in six geriatric wards. We studied 169 patients with varying degrees of hydration: dehydrated, euvolemic, and overhydrated. BIA estimates of TBW and of ECW were compared with the measurement of TBW with (18)O dilution and of ECW with bromide (Br) dilution. RESULTS: BIA estimated TBW with a difference of 0.48 +/- 2.3 l (mean +/- SD) (50 kHz; p = .01) and 0.69 +/- 2.2 l (100 kHz; p < 0.001) compared with (18)O dilution. The difference was not affected by the hydration status. Estimates of ECW with BIA were systematically biased compared with Br dilution: 4.6 +/- 3.1 l (equation from Segal and colleagues; p < .001) and 3.4 +/- 2.9 l (equation from Visser and colleagues; p < .001). We propose a new, cross-validated equation. Conclusions. Body water spaces can be estimated accurately in geriatric patients with BIA. PMID- 11382793 TI - Chronic cellular dehydration in the aged patient. AB - BACKGROUND: As a reduction of water spaces is expected in the elderly because of fat-free mass loss, disease is often associated with increased hydration. The present study compared water spaces and cellular hydration in adults, healthy and diseased aged patients. METHODS: An open study was conducted in 6 geriatric wards and a nutrition laboratory involving 85 aged diseased persons, 68 healthy elderly adults, and 35 adults. Total body water (TBW, H(2)(18)O dilution), extracellular water (ECW, Bromide dilution), and fat-free mass (FFM, body density and Siri's equation) were measured directly whereas intracellular water (ICW = TBW - ECW) and body cell mass (FFM - ECW) were obtained by calculations. RESULTS: FFM, TBW, and ICW were higher in adults than in the 2 other groups and in the elderly than in aged patients. ECW was higher in aged patients than in healthy elderly participants. The proportion of TBW made of ECW or ICW was the same in adults and in healthy elderly persons. A higher proportion of TBW was composed of ECW, and a lower proportion of TBW was composed of ICW, in diseased patients compared with the 2 other groups. The proportion of ICW in body cell mass was also lower in diseased patients. CONCLUSIONS: Diseased elderly persons display reduced ICW and expanded ECW. A cellular dehydration is suggested. PMID- 11382794 TI - Endogenous anticholinergic substances may exist during acute illness in elderly medical patients. AB - BACKGROUND: The purpose of this study was to determine if serum anticholinergic activity (SACA) arises from endogenous substances produced during illness. METHODS: Elderly medical inpatients (N = 612) were screened for anticholinergic medication use in the week prior to the study by interviews of subjects and proxies and review of emergency room, hospital, and nursing home medication administration records. Of 24 subjects without a recent anticholinergic medication history, 15 were recruited and 10 completed the study. Serum samples were obtained on Day 2 of hospital admission. SACA was measured using a radionuclide displacement assay. Medications taken by subjects were assayed for central muscarinic receptor binding at therapeutic concentrations. Results. Eight of the ten subjects had SACA detectable in the serum. No medication used by these subjects had anticholinergic activity at usual therapeutic concentrations. CONCLUSIONS: Endogenous anticholinergic substances may exist during acute illness. Characterization of such substances may increase the depth of our understanding of delirium and lead to useful intervention strategies. PMID- 11382795 TI - Bright light treatment decreases depression in institutionalized older adults: a placebo-controlled crossover study. AB - BACKGROUND: An important parallel exists between patients with seasonal affective disorder and institutionalized older adults. Many older patients, as a result of global physical decline and immobility, are confined to their rooms, experiencing little natural sunlight. Thus, institutionalized older adults are at risk for chronic light deprivation. Testing the hypothesis that chronic light deprivation might be responsible, at least in part, for some depression among institutionalized older adults, the aim of this study was to investigate the efficacy of morning bright light treatment on depression among older adults residing in a long-term care facility. METHODS: In a placebo controlled, crossover design, participants (N = 10, six women and four men; M age = 83.8) received each of the following: (i) 1 week (5 days) of 10,000 lux (therapeutic dose); (ii) 1 week (5 days) of 300 lux (placebo); or 1 week of no treatment (control). Each week of light treatment was 5 consecutive days, 30 minutes daily, with a wash-out period consisting of 1 week between conditions. RESULTS: Geriatric Depression Scale (GDS) scores at baseline during all treatment conditions were positively correlated (r = .81, p < .01) with months of institutionalization, where participants with higher GDS scores experienced more time institutionalized. Scores on the GDS remained unchanged during the placebo and control conditions, but depression scores decreased significantly during the 10,000 lux treatment (pretest GDS M = 15 vs posttest GDS M = 11, p < .01). After the 10,000 lux treatment, 50% of the participants no longer scored in the depressed range. Improvement during the 10,000 lux condition was positively correlated (r = .62, p < .05) to baseline GDS scores, where participants with higher GDS scores experienced greater improvement following the 10,000 lux treatment. CONCLUSIONS: The results of the present study suggest that bright light treatment may be effective among institutionalized older adults, providing nonpharmacological intervention in the treatment of depression. Furthermore, the length of institutionalization may play an important role in determining the efficacy of bright light treatment for older adults in the nursing-home setting. PMID- 11382796 TI - Reasons prompting digitalis therapy in the acute care hospital. AB - BACKGROUND: The choice of administering digitalis to older patients with congestive heart failure (CHF) cannot be made on the account of univocally defined criteria because of uncertainty about efficacy and concern about safety of digitalis in this population. The purpose of this study was to verify whether the clinical characteristics on admission to the acute care hospital determine the use of digitalis therapy in elderly patients. METHODS: A total of 1239 patients (mean age 77.8 +/- 7.1 years, range 65-100 years, males 49.8%) consecutively admitted to 69 General Medicine and Geriatrics wards over a 4-month period were grouped by combining two dichotomous factors (Carlson's score > 4: definite or possible diagnosis of CHF; Carlson's score < 5: unlikely diagnosis of CHF; in-hospital adoption of digitalis therapy: yes or no) as follows: Group A: Carlson's score > 4, digitalis (n = 413); Group B: Carlson's score > 4, no digitalis (n = 260); Group C: Carlson's score < 5, digitalis (n = 104); Group D: Carlson's score < 5, no digitalis (n = 462). Variables significantly distinguishing groups were entered into a discriminant analysis aimed at assessing the group specificity of individual clinical profiles. RESULTS: Use of digoxin at home, atrial fibrillation, older age, and comorbidity (mainly COPD and chronic renal failure) characterized most of the patients given digoxin with or without a definite diagnosis of CHF. Clinical profiles of groups A, B, and C largely overlapped. CONCLUSION: Age, historical use of digitalis, and comorbidity might lead to seemingly incongruous digitalis prescription. The choice of adopting digitalis therapy cannot be reliably predicted on the basis of clinical variables only. Presently unexplored physician-related factors, such as cultural background, likely outweigh clinical variables in prompting digitalis prescription. PMID- 11382797 TI - Screening for undernutrition in geriatric practice: developing the short-form mini-nutritional assessment (MNA-SF). AB - BACKGROUND: The Mini-Nutritional Assessment (MNA) is a validated assessment instrument for nutritional problems, but its length limits its usefulness for screening. We sought to develop a screening version of this instrument, the MNA SF, that retains good diagnostic accuracy. METHODS: We reanalyzed data from France that were used to develop the original MNA and combined these with data collected in Spain and New MEXICO: Of the 881 subjects with complete MNA data, 151 were from France, 400 were from Spain, and 330 were from New MEXICO: Independent ratings of clinical nutritional status were available for 142 of the French subjects. Overall, 73.8% were community dwelling, and mean age was 76.4 years. Items were chosen for the MNA-SF on the basis of item correlation with the total MNA score and with clinical nutritional status, internal consistency, reliability, completeness, and ease of administration. RESULTS: After testing multiple versions, we identified an optimal six-item MNA-SF total score ranging from 0 to 14. The cut-point score for MNA-SF was calculated using clinical nutritional status as the gold standard (n = 142) and using the total MNA score (n = 881). The MNA-SF was strongly correlated with the total MNA score (r = .945). Using an MNA-SF score of > or = 11 as normal, sensitivity was 97.9%, specificity was 100%, and diagnostic accuracy was 98.7% for predicting undernutrition. CONCLUSIONS: The MNA-SF can identify persons with undernutrition and can be used in a two-step screening process in which persons, identified as "at risk" on the MNA-SF, would receive additional assessment to confirm the diagnosis and plan interventions. PMID- 11382798 TI - The recommended dietary allowance for protein may not be adequate for older people to maintain skeletal muscle. AB - BACKGROUND: Inadequate dietary protein intake results in loss of skeletal muscle mass. Some shorter-term nitrogen balance studies suggest that the Recommended Dietary Allowance (RDA) of protein may not be adequate for older people. The aim of this study was to assess the adequacy of the RDA of protein for older people by examining longer-term responses in urinary nitrogen excretion, whole-body protein metabolism, whole-body composition, and mid-thigh muscle area. METHODS: This was a 14-week precisely controlled diet study. Ten healthy, ambulatory men and women, aged 55 to 77 years, were provided eucaloric diets that contained 0.8 g protein.kg(-1).day(-1). The study was conducted at a General Clinical Research Center using an outpatient setting for 11 weeks and an inpatient setting for 3 weeks. The main outcome measures included urinary nitrogen excretion, postabsorptive and postprandial whole-body leucine kinetics via infusion of L-[1 (13)C]-leucine, whole-body density via hydrostatic weighing, total body water via deuterium oxide dilution, and mid-thigh muscle area via computed tomography scans. RESULTS: Mean urinary nitrogen excretion decreased over time from Weeks 2 to 8 to 14 (p =.025). At Week 14, compared with Week 2, there were no changes in postabsorptive or postprandial leucine kinetics (turnover, oxidation, incorporation into protein via synthesis, release via breakdown, or balance). Whole-body composition (% body fat, fat-free mass, and protein + mineral mass) did not change over time in these weight-stable subjects. Mid-thigh muscle area was decreased by -1.7 +/- 0.6 cm(2) (p =.019) at Week 14 compared with Week 2. The loss of mid-thigh muscle area was associated with the decrease in urinary nitrogen excretion (Spearman r =.83, p =.010). CONCLUSIONS: The maintenance of whole-body leucine metabolism and whole-body composition is generally consistent with a successful adaptation to the RDA for protein. However, the decrease in mid thigh muscle area and the association with decreased urinary nitrogen excretion are consistent with a metabolic accommodation. These results suggest that the RDA for protein may not be adequate to completely meet the metabolic and physiological needs of virtually all older people. PMID- 11382799 TI - Differences in the neuromuscular capacity and lean muscle tissue in old and older community-dwelling adults. AB - BACKGROUND: This study investigated whether there was a worsening of the neuromuscular capacity of older adults after the seventh decade of life. METHODS: Fifteen healthy community-dwelling old (< 70 years of age) and 15 older adults (> or = 70 years of age) were assessed for maximal isometric strength (MVC) and force production characteristics, a one-repetition maximum (1-RM) performance, electromyographic (EMG) activity, and bone-free lean tissue (BFLT) mass of the lower extremity. RESULTS: The isometric MVC, 1-RM, and BFLT mass values in the old group were significantly greater than in the older group. In addition, the individual BFLT mass values correlated significantly with the isometric MVC values (r = .85) and the 1-RM scores of the thigh muscle groups (r = .54-.80). The old group generated significantly greater isometric maximal rate of torque development than the older group and performed significantly better at all intervals of the absolute and relative force-time curves. The voluntary muscle activation of the knee extensors of the old group produced significantly higher integrated EMG (iEMG) activity at each epoch in the early iEMG-time curve compared with the old group. CONCLUSIONS: The results suggest that the age related deterioration in maximal strength measures and rapid force production characteristics in older adults could be related to a reduction in the mass and neural activation of the thigh muscles. The deterioration of the neuromuscular system of community-dwelling older adults may contribute to an increased difficulty in performing daily activities and may increase their risks of tripping and falling. PMID- 11382800 TI - Effect of age on ocular microtremor activity. AB - BACKGROUND: Ocular microtremor (OMT) is a high-frequency tremor of the eyes. It is present in all individuals and is related to brainstem activity. The OMT signal appears as an irregular oscillatory movement with intermittent burst-like components. The clinical interest in OMT has centered on its use in the assessment of the comatose patient, with broad agreement among authors of its prognostic value. The purpose of this study was to examine the changes in OMT activity related to aging. METHODS: OMT was recorded from 72 normal healthy subjects using the piezoelectric strain gauge technique. The subjects ranged in age from 21 to 88 years (54.22 +/- 20.43 years, mean +/- SD). RESULTS: Our results show that the overall frequency and frequency content of the bursts falls with age (p < .002 and p < .001, respectively). There is a highly significant drop in all three frequency parameters of OMT (p < .0001) in subjects older than 60 years of age. CONCLUSIONS: These results suggest that different values of normality should operate for subjects over 60 years of age when considering the clinical application of OMT. PMID- 11382801 TI - Efficacy of Influenza Vaccine in Elderly Persons in Welfare Nursing Homes: Reduction in Risks of Mortality and Morbidity During an Influenza A (H3N2) Epidemic. AB - BACKGROUND: The effect of influenza vaccination on the occurrence and severity of influenza virus infection in a population residing in nursing homes was studied through a program by the Osaka Prefectural Government, which is the first and official support for influenza vaccination of the elderly population during an influenza A (H3N2) epidemic in JAPAN: METHODS: A cohort study located in the Osaka Prefecture, Japan, followed the outcomes of elderly nursing home residents who received influenza vaccinations (n = 10,739) in comparison with control subjects who did not receive influenza vaccinations (n = 11,723) and monitored clinically the onset of serious morbidity and mortality of influenza illness. Subjects were 22,462 persons older than 65 years who resided in 301 welfare nursing homes in the Osaka Prefecture, Japan during an influenza A (H3N2) epidemic in 1998 to 1999. RESULTS: Of 22,462 individuals living in 301 nursing homes, 10,739 received either one dose (2027 subjects) or two doses (8712 subjects) of inactivated, subunit trivalent influenza vaccine. Through the period from November 1998 to March 1999, there were 950 cases of influenza infection diagnosed clinically with cases by virus isolation and/or serology. There were statistically significantly fewer clinical cases of influenza, hospital admissions due to severe infection, and deaths due to influenza in the vaccinated cohort (256 cases, 32 hospital admissions, and one death) compared with the unvaccinated controls (694 cases, 150 hospital admissions, and five deaths). Vaccination was equally effective in those who received one dose of vaccine as in those who received two doses. No serious adverse reactions to vaccination were recorded. Thus, influenza vaccination is safe and effective in this population and should be an integral part of the routine care of persons aged 65 years and older residing in nursing homes. CONCLUSIONS: This study provides an analysis of the clinical efficacy of influenza vaccination in a large cohort of nursing home residents in JAPAN: Annual influenza vaccine administration requires the attention of all nursing home attendants, physicians, and public health organizations. PMID- 11382802 TI - Interaction of scorpion alpha-toxins with cardiac sodium channels: binding properties and enhancement of slow inactivation. AB - The effects of the scorpion alpha-toxins Lqh II, Lqh III, and LqhalphaIT on human cardiac sodium channels (hH1), which were expressed in human embryonic kidney (HEK) 293 cells, were investigated. The toxins removed fast inactivation with EC(50) values of <2.5 nM (Lqh III), 12 nM (Lqh II), and 33 nM (LqhalphaIT). Association and dissociation rates of Lqh III were much slower than those of Lqh II and LqhalphaIT, such that Lqh III would not dissociate from the channel during a cardiac activation potential. The voltage dependence of toxin dissociation from hH1 channels was nearly the same for all toxins tested, but it was different from that found for skeletal muscle sodium channels (muI; Chen et al. 2000). These results indicate that the voltage dependence of toxin binding is a property of the channel protein. Toxin dissociation remained voltage dependent even at high voltages where activation and fast inactivation is saturated, indicating that the voltage dependence originates from other sources. Slow inactivation of hH1 and muI channels was significantly enhanced by Lqh II and Lqh III. The half-maximal voltage of steady-state slow inactivation was shifted to negative values, the voltage dependence was increased, and, in particular for hH1, slow inactivation at high voltages became more complete. This effect exceeded an expected augmentation of slow inactivation owing to the loss of fast inactivation and, therefore, shows that slow sodium channel inactivation may be directly modulated by scorpion alpha-toxins. PMID- 11382803 TI - Integrated allosteric model of voltage gating of HCN channels. AB - Hyperpolarization-activated (pacemaker) channels are dually gated by negative voltage and intracellular cAMP. Kinetics of native cardiac f-channels are not compatible with HH gating, and require closed/open multistate models. We verified that members of the HCN channel family (mHCN1, hHCN2, hHCN4) also have properties not complying with HH gating, such as sigmoidal activation and deactivation, activation deviating from fixed power of an exponential, removal of activation "delay" by preconditioning hyperpolarization. Previous work on native channels has indicated that the shifting action of cAMP on the open probability (Po) curve can be accounted for by an allosteric model, whereby cAMP binds more favorably to open than closed channels. We therefore asked whether not only cAMP-dependent, but also voltage-dependent gating of hyperpolarization-activated channels could be explained by an allosteric model. We hypothesized that HCN channels are tetramers and that each subunit comprises a voltage sensor moving between "reluctant" and "willing" states, whereas voltage sensors are independently gated by voltage, channel closed/open transitions occur allosterically. These hypotheses led to a multistate scheme comprising five open and five closed channel states. We estimated model rate constants by fitting first activation delay curves and single exponential time constant curves, and then individual activation/deactivation traces. By simply using different sets of rate constants, the model accounts for qualitative and quantitative aspects of voltage gating of all three HCN isoforms investigated, and allows an interpretation of the different kinetic properties of different isoforms. For example, faster kinetics of HCN1 relative to HCN2/HCN4 are attributable to higher HCN1 voltage sensors' rates and looser voltage-independent interactions between subunits in closed/open transitions. It also accounts for experimental evidence that reduction of sensors' positive charge leads to negative voltage shifts of Po curve, with little change of curve slope. HCN voltage gating thus involves two processes: voltage sensor gating and allosteric opening/closing. PMID- 11382804 TI - Cysteine mutagenesis reveals novel structure-function features within the predicted third extracellular loop of the type IIa Na(+)/P(i) cotransporter. AB - The transport function of the rat type IIa Na(+)/P(i) cotransporter is inhibited after binding the cysteine modifying reagent 2-aminoethyl methanethiosulfonate hydrobromide (MTSEA) to a cysteine residue substituted for a serine at position 460 (S460C) in the predicted third extracellular loop. This suggests that Ser-460 lies in a functionally important region of the protein. To establish a "structure function" profile for the regions that flank Ser-460, the substituted cysteine accessibility method was employed. 18 mutants were constructed in which selected amino acids from Arg-437 through Leu-465 were substituted one by one for a cysteine. Mutants were expressed in Xenopus oocytes and transport function (cotransport and slippage) and kinetics were assayed by electrophysiology with or without prior treatment with cysteine modifying (methanethiosulfonate, MTS) reagents. Except for mutant I447C, mutants with cysteines at sites from Arg-437 through Thr-449, as well as Pro-461, were inactive. Cotransport function of mutants with Cys substitutions at sites Arg-462 through Leu-465 showed low sensitivity to MTS reagents. The preceding mutants (Cys substitution at Thr-451 to Ser-460) showed a periodic accessibility pattern that would be expected for an alpha-helix motif. Apart from loss of transport function, exposure of mutants A453C and A455C to MTSEA or 2-(triethylammonium)ethyl MTS bromide (MTSET) increased the uncoupled slippage current, which implicated the mutated sites in the leak pathway. Mutants from Ala-453 through Ala-459 showed less pH dependency, but generally stronger voltage dependency compared with the wild type, whereas those flanking this group were more sensitive to pH and showed weaker voltage dependence of cotransport mode kinetics. Our data indicate that parts of the third extracellular loop are involved in the translocation of the fully loaded carrier and show a membrane-associated alpha-helical structure. PMID- 11382805 TI - Early intermediates in the transport cycle of the neuronal excitatory amino acid carrier EAAC1. AB - Electrogenic glutamate transport by the excitatory amino acid carrier 1 (EAAC1) is associated with multiple charge movements across the membrane that take place on time scales ranging from microseconds to milliseconds. The molecular nature of these charge movements is poorly understood at present and, therefore, was studied in this report in detail by using the technique of laser-pulse photolysis of caged glutamate providing a 100-micros time resolution. In the inward transport mode, the deactivation of the transient component of the glutamate induced coupled transport current exhibits two exponential components. Similar results were obtained when restricting EAAC1 to Na(+) translocation steps by removing potassium, thus, demonstrating (1) that substrate translocation of EAAC1 is coupled to inward movement of positive charge and, therefore, electrogenic; and (2) the existence of at least two distinct intermediates in the Na(+)-binding and glutamate translocation limb of the EAAC1 transport cycle. Together with the determination of the sodium ion concentration and voltage dependence of the two exponential charge movement and of the steady-state EAAC1 properties, we developed a kinetic model that is based on sequential binding of Na(+) and glutamate to their extracellular binding sites on EAAC1 explaining our results. In this model, at least one Na(+) ion and thereafter glutamate rapidly bind to the transporter initiating a slower, electroneutral structural change that makes EAAC1 competent for further, voltage-dependent binding of additional sodium ion(s). Once the fully loaded EAAC1 complex is formed, it can undergo a much slower, electrogenic translocation reaction to expose the substrate and ion binding sites to the cytoplasm. PMID- 11382806 TI - Single-channel properties of recombinant acid-sensitive ion channels formed by the subunits ASIC2 and ASIC3 from dorsal root ganglion neurons expressed in Xenopus oocytes. AB - The acid-sensitive ion channels known as ASIC are gated by external protons. A set of these channels is expressed in dorsal root ganglion neurons where they may participate in the transduction of mechanical and nociceptive stimuli. Here, we have examined the single-channel properties of channels formed by the subunits ASIC2 and ASIC3 expressed in Xenopus oocytes using outside-out patches. The mean single-channel current-voltage relationship is linear with a slope conductance of 18 pS between -80 and -40 mV in 150 mM Na(+) outside and 150 mM K(+) inside the patch pipet. The selectivity for monovalent cations has the sequence Na(+) > Li(+) > K(+). Divalent cations such as Ca(2+) do not permeate, but instead block the channel when applied to the extracellular side. External protons increase the probability of channels being open to a maximum of 0.8 with an EC(50) of 16 +/- 4 microM and a Hill coefficient of 2.7 +/- 0.3, whereas the mean single-channel current amplitude is independent of external pH. Analysis of the kinetics of single channels indicates the presence of at least four modes of activity (Mod1 to Mod4) in addition to an inactivated state. Three of the modes exhibit distinct kinetics, and can be unambiguously identified on the basis of open probability (P(oMod1) = 0.5 +/- 0.05; P(oMod2) > 0.9 +/- 0.05; P(oMod3) < 0.1). Mode 4, which has a P(o) in the range of 0.5-0.8, may constitute a distinct mode or alternatively, it represents transitions between the other three modes of activity. Increasing [H(+)](o) increases the frequency of entering the modes with high P(o) (modes 1, 2, and 4) and the time the channel spends in the modes with high activity. PMID- 11382807 TI - Role of aquaporin water channels in airway fluid transport, humidification, and surface liquid hydration. AB - Several aquaporin-type water channels are expressed in mammalian airways and lung: AQP1 in microvascular endothelia, AQP3 in upper airway epithelia, AQP4 in upper and lower airway epithelia, and AQP5 in alveolar epithelia. Novel quantitative methods were developed to compare airway fluid transport-related functions in wild-type mice and knockout mice deficient in these aquaporins. Lower airway humidification, measured from the moisture content of expired air during mechanical ventilation with dry air through a tracheotomy, was 54-56% efficient in wild-type mice, and reduced by only 3-4% in AQP1/AQP5 or AQP3/AQP4 double knockout mice. Upper airway humidification, measured from the moisture gained by dry air passed through the upper airways in mice breathing through a tracheotomy, decreased from 91 to 50% with increasing ventilation from 20 to 220 ml/min, and reduced by 3-5% in AQP3/AQP4 knockout mice. The depth and salt concentration of the airway surface liquid in trachea was measured in vivo using fluorescent probes and confocal and ratio imaging microscopy. Airway surface liquid depth was 45 +/- 5 microm and [Na(+)] was 115 +/- 4 mM in wild-type mice, and not significantly different in AQP3/AQP4 knockout mice. Osmotic water permeability in upper airways, measured by an in vivo instillation/sample method, was reduced by approximately 40% by AQP3/AQP4 deletion. In doing these measurements, we discovered a novel amiloride-sensitive isosmolar fluid absorption process in upper airways (13% in 5 min) that was not affected by aquaporin deletion. These results establish the fluid transporting properties of mouse airways, and indicate that aquaporins play at most a minor role in airway humidification, ASL hydration, and isosmolar fluid absorption. PMID- 11382808 TI - Inactivation of BK channels mediated by the NH(2) terminus of the beta3b auxiliary subunit involves a two-step mechanism: possible separation of binding and blockade. AB - A family of auxiliary beta subunits coassemble with Slo alpha subunit to form Ca(2)+-regulated, voltage-activated BK-type K(+) channels. The beta subunits play an important role in regulating the functional properties of the resulting channel protein, including apparent Ca(2)+ dependence and inactivation. The beta3b auxiliary subunit, when coexpressed with the Slo alpha subunit, results in a particularly rapid ( approximately 1 ms), but incomplete inactivation, mediated by the cytosolic NH(2) terminus of the beta3b subunit (Xia et al. 2000). Here, we evaluate whether a simple block of the open channel by the NH(2)-terminal domain accounts for the inactivation mechanism. Analysis of the onset of block, recovery from block, time-dependent changes in the shape of instantaneous current-voltage curves, and properties of deactivation tails suggest that a simple, one step blocking reaction is insufficient to explain the observed currents. Rather, blockade can be largely accounted for by a two-step blocking mechanism (C(n) <-- > O(n) <---> O(*)(n) <---> I(n)) in which preblocked open states (O*(n)) precede blocked states (I(n)). The transitions between O* and I are exceedingly rapid accounting for an almost instantaneous block or unblock of open channels observed with changes in potential. However, the macroscopic current relaxations are determined primarily by slower transitions between O and O*. We propose that the O to O* transition corresponds to binding of the NH(2)-terminal inactivation domain to a receptor site. Blockade of current subsequently reflects either additional movement of the NH(2)-terminal domain into a position that hinders ion permeation or a gating transition to a closed state induced by binding of the NH(2) terminus. PMID- 11382809 TI - Gating properties conferred on BK channels by the beta3b auxiliary subunit in the absence of its NH(2)- and COOH termini. AB - Both beta1 and beta2 auxiliary subunits of the BK-type K(+) channel family profoundly regulate the apparent Ca(2)+ sensitivity of BK-type Ca(2)+-activated K(+) channels. Each produces a pronounced leftward shift in the voltage of half activation (V(0.5)) at a given Ca(2)+ concentration, particularly at Ca(2)+ above 1 microM. In contrast, the rapidly inactivating beta3b auxiliary produces a leftward shift in activation at Ca(2)+ below 1 microM. In the companion work (Lingle, C.J., X.-H. Zeng, J.-P. Ding, and X.-M. Xia. 2001. J. Gen. Physiol. 117:583-605, this issue), we have shown that some of the apparent beta3b-mediated shift in activation at low Ca(2)+ arises from rapid unblocking of inactivated channels, unlike the actions of the beta1 and beta2 subunits. Here, we compare effects of the beta3b subunit that arise from inactivation, per se, versus those that may arise from other functional effects of the subunit. In particular, we examine gating properties of the beta3b subunit and compare it to beta3b constructs lacking either the NH(2)- or COOH terminus or both. The results demonstrate that, although the NH(2) terminus appears to be the primary determinant of the beta3b-mediated shift in V(0.5) at low Ca(2)+, removal of the NH(2) terminus reveals two other interesting aspects of the action of the beta3b subunit. First, the conductance-voltage curves for activation of channels containing the beta3b subunit are best described by a double Boltzmann shape, which is proposed to arise from two independent voltage-dependent activation steps. Second, the presence of the beta3b subunit results in channels that exhibit an anomalous instantaneous outward current rectification that is correlated with a voltage dependence in the time-averaged single-channel current. The two effects appear to be unrelated, but indicative of the variety of ways that interactions between beta and alpha subunits can affect BK channel function. The COOH terminus of the beta3b subunit produces no discernible functional effects. PMID- 11382810 TI - Genes encoding the vacuolar Na+/H+ exchanger and flower coloration. AB - Vacuolar pH plays an important role in flower coloration: an increase in the vacuolar pH causes blueing of flower color. In the Japanese morning glory (Ipomoea nil or Pharbitis nil), a shift from reddish-purple buds to blue open flowers correlates with an increase in the vacuolar pH. We describe details of the characterization of a mutant that carries a recessive mutation in the Purple (Pr) gene encoding a vacuolar Na+/H+ exchanger termed InNHX1. The genome of I. nil carries one copy of the Pr (or InNHX1) gene and its pseudogene, and it showed functional complementation to the yeast nhx1 mutation. The mutant of I. nil, called purple (pr), showed a partial increase in the vacuolar pH during flower opening and its reddish-purple buds change into purple open flowers. The vacuolar pH in the purple open flowers of the mutant was significantly lower than that in the blue open flowers. The InNHX1 gene is most abundantly expressed in the petals at around 12 h before flower-opening, accompanying the increase in the vacuolar pH for the blue flower coloration. No such massive expression was observed in the petunia flowers. Since the NHX1 genes that promote the transport of Na+ into the vacuoles have been regarded to be involved in salt tolerance by accumulating Na+ in the vacuoles, we can add a new biological role for blue flower coloration in the Japanese morning glory by the vacuolar alkalization. PMID- 11382811 TI - A large family of class III plant peroxidases. AB - Class III plant peroxidase (POX), a plant-specific oxidoreductase, is one of the many types of peroxidases that are widely distributed in animals, plants and microorganisms. POXs exist as isoenzymes in individual plant species, and each isoenzyme has variable amino acid sequences and shows diverse expression profiles, suggesting their involvement in various physiological processes. Indeed, studies have provided evidence that POXs participate in lignification, suberization, auxin catabolism, wound healing and defense against pathogen infection. Little, however, is known about the signal transduction for inducing expression of the pox genes. Recent studies have provided information on the regulatory mechanisms of wound- and pathogen-induced expression of some pox genes. These studies suggest that pox genes are induced via different signal transduction pathways from those of other known defense-related genes. PMID- 11382812 TI - Chlorophyll degradation in a Chlamydomonas reinhardtii mutant: an accumulation of pyropheophorbide a by anaerobiosis. AB - Chlorophyll degradation was investigated in cells of a chlorophyll b-less mutant of Chlamydomonas reinhardtii under aerobic and anaerobic conditions. During degradation of chlorophyll under anaerobic conditions, chlorophyll catabolite P535, an open-tetrapyrrole, was not excreted, but pyropheophorbide a was accumulated as the end product with a transient accumulation of chlorophyllide a and pheophorbide a in cells, in contrast to the breakdown under aerobic conditions. It is likely that in the absence of oxygen, degradation of chlorophyll a proceeds to pyropheophorbide a by three consecutive reactions, dephytylation, metal-releasing and demethoxycarbonylation, and then stops due to a limitation of the oxygen that the monooxygenase reaction requires for bilin formation. A novel enzyme catalyzing demethoxycarbonylation of pheophorbide a was partially purified. The enzyme activity increased dependent on the age of cells, and its increase was completely suppressed by cycloheximide. Production of P535 was also dependent on cytoplasmic protein synthesis. PMID- 11382813 TI - Cloning and characterization of a hydroxycinnamoyl-CoA:tyramine N (hydroxycinnamoyl)transferase induced in response to UV-C and wounding from Capsicum annuum. AB - Hydroxycinnamoyl-CoA : tyramine N-(hydroxycinnamoyl) transferase (THT) is a pivotal enzyme in the synthesis of N-(hydroxycinnamoyl)-amines, which are associated with cell wall fortification in plants. The cDNA encoding THT was cloned from the leaves of UV-C treated Capsicum annuum (hot pepper) using a differential screening strategy. The predicted protein encoded by the THT cDNA is 250 amino acids in length and has a relative molecular mass of 28,221. The protein sequence derived from the cDNA shares 76% and 67% identity with the potato and tobacco THT protein sequences, respectively. The recombinant pepper THT enzyme was purified using a bacterial overexpression system. The purified enzyme has a broad substrate specificity including acyl donors such as cinnamoyl , sinapoyl-, feruloyl-, caffeoyl-, and 4-coumaroyl-CoA and acceptors such as tyramine and octopamine. In UV-C treated plants, the THT mRNA was strongly induced in leaves, and the elevated level of expression was stable for up to 36 h. THT mRNA also increased in leaves that were detached from the plant but not treated with UV-C. THT expression was measured in different plant tissues, and was constitutive at a similar level in leaf, root, stem, flower and fruit. Induction of THT mRNA was correlated with an increase in THT protein. PMID- 11382814 TI - Absence of lutein, violaxanthin and neoxanthin affects the functional chlorophyll antenna size of photosystem-II but not that of photosystem-I in the green alga Chlamydomonas reinhardtii. AB - Chlamydomonas reinhardtii double mutant npq2 lor1 lacks the beta, epsilon carotenoids lutein and loroxanthin as well as all beta,beta-epoxycarotenoids derived from zeaxanthin (e.g. violaxanthin and neoxanthin). Thus, the only carotenoids present in the thylakoid membranes of the npq2 lor1 cells are beta carotene and zeaxanthin. The effect of these mutations on the photochemical apparatus assembly and function was investigated. In cells of the mutant strain, the content of photosystem-II (PSII) and photosystem-I (PSI) was similar to that of the wild type, but npq2 lor1 had a significantly smaller PSII light-harvesting Chl antenna size. In contrast, the Chl antenna size of PSI was not truncated in the mutant. SDS-PAGE and Western blot analysis qualitatively revealed the presence of all LHCII and LHCI apoproteins in the thylakoid membrane of the mutant. The results showed that some of the LHCII and most of the LHCI were assembled and functionally connected with PSII and PSI, respectively. Photon conversion efficiency measurements, based on the initial slope of the light saturation curve of photosynthesis and on the yield of Chl a fluorescence in vivo, showed similar efficiencies. However, a significantly greater light intensity was required for the saturation of photosynthesis in the mutant than in the wild type. It is concluded that zeaxanthin can successfully replace lutein and violaxanthin in most of the functional light-harvesting antenna of the npq2 lor1 mutant. PMID- 11382815 TI - Molecular cloning of beta-galactosidase from Japanese pear (Pyrus pyrifolia) and its gene expression with fruit ripening. AB - We have cloned a cDNA fragment encoding a beta-galactosidase from Japanese pear (Pyrus pyrifolia) fruit (JP-GAL). It contained an untranslated sequence of 182 nucleotides at the 5' end, a presumptive coding sequence of 2,193 nucleotides and an untranslated sequence of 268 nucleotides including a polyadenylation signal and a poly (A) tail at the 3' end. It encoded a protein with a calculated molecular weight of 80.9 kDa which consists of 731 amino acids. Both the nucleotide and the deduced amino acid sequences showed a 98% sequence identity with that obtained from the apple beta-galactosidase cDNA. The peptide sequence obtained from the purified Japanese pear beta-galactosidase III matched the deduced amino acid sequence of SVSYDHKAIIINGQKRILISG (amino acid 25-45). Northern blot analysis showed that the probe derived from JP-GAL hybridized to a single 2.6 kb RNA. The mRNA was detected solely in the fruit; none was detected in the buds, leaves, roots or shoots of the Japanese pear. The steady-state level of the beta-galactosidase mRNA was measured during fruit ripening in three cultivars, Housui, Kousui (early ripening) and Niitaka (late ripening). The results showed that regardless of the cultivar, no JP-GAL mRNA was detected in the immature fruit. Increment of the mRNA level with fruit ripening coincided with the increase in the beta-galactosidase III activity. Our results showed that the expression of JP-GAL correlated with fruit softening and JP-GAL may be beta galactosidase III. PMID- 11382816 TI - EMF genes interact with late-flowering genes in regulating floral initiation genes during shoot development in Arabidopsis thaliana. AB - To investigate the mechanisms regulating the initiation of floral development in Arabidopsis, a construct containing beta-glucuronidase (GUS) gene driven by APETALA1 promoter (AP1::GUS) was introduced into emf fwa and emf ft double mutants. GUS activity was strongly detected on shoot meristem of emf1-1 single mutants harboring AP1::GUS construct just 5 d after germination. By contrast, GUS activity was undetectable on emf1-1 fwa-1, emf1-1 ft-1, emf2-1 fwa-1, emf2-3 fwa 1 and emf2-3 ft-1 double mutants harboring AP1::GUS construct 10 d after germination. GUS activity was only weakly detected on the apical meristem of 20 day-old emf1-1 fwa-1 and emf2-1 fwa-1 seedlings. During this time, only sessile leaves were produced. Further analysis indicated that AP1 was strongly expressed in 10-day-old emf1-1 and emf2-1 single mutants. Its expression was significantly reduced in all emf1-1 or emf2-1 late-flowering double mutants tested. Similar to AP1, the expression of LEAFY (LFY) was also high in emf1-1 and emf2-1 single mutants and reduced in emf1-1 or emf2-1 late-flowering double mutants. Our results indicate that the precocious expression of AP1 and LFY is dependent not only on the low EMF and FWA activities but also on the expression of most of the late-flowering genes such as FT, FCA, FE, CO and GI. These data also reveal that most late-flowering genes may function downstream of EMF or in pathways distinct from EMF to activate genes specified floral meristem identity during shoot maturation in Arabidopsis. PMID- 11382817 TI - Cyclic flow of electrons within PSII in thylakoid membranes. AB - In photosynthesis, the electrons released from PSII are considered to be shared mainly by carbon metabolism and the water-water cycle. We demonstrated previously that some electrons are utilized in a CO2- and O2-independent manner in leaves of wild watermelon [Miyake and Yokota (2000) Plant Cell Physiol: 41: 335]. In the present study, we examined the mechanism of this alternative flow of electrons in thylakoid membranes, isolated from fresh spinach leaves, by simultaneously measuring the quantum yield of PSII and the flux of the linear flow of electrons. In the presence of the protonophore nigericin, which eliminates the pH gradient across thylakoid membranes, the quantum yield and the flux of the linear flow of electrons were directly proportional to one another. The quantum yield at a given linear flux of electrons was much higher in the absence of nigericin than in its presence, indicating that an additional or alternative flow of electrons can occur independently of the linear flow in the absence of nigericin. In the presence of nigericin, the alternative flux of electrons increased with decreasing pH and with increasing reduction of the plastoquinone pool. Cyclic flow of electrons in PSII appears to be the most plausible candidate for the alternative flow of electrons. The flux reached 280 micromol x e(-) (mg Chl)(-1) x h(-1) and was similar to that of the CO2- and O2-independent alternative flow of electrons that we found in leaves of wild watermelon. The cyclic, alternative flow of electrons in PSII provides a possible explanation for the alternative flow of electrons observed in vivo. PMID- 11382818 TI - Modulation in the activity of purified tonoplast H+-ATPase by tonoplast glycolipids prepared from cultured rice (Oryza sativa L. var. Boro) cells. AB - Glycolipids, phospholipids, and neutral lipids were extracted from the tonoplast fraction of cultured rice cells (Oryza sativa L. var. Boro). Acyl steryl glucoside (ASG) and glucocerebroside (GlcCer) were also prepared from this fraction. We determined the effects of these tonoplast lipids on the activity of H+-ATPase which was delipidated and purified from the tonoplast fraction. Exogenously added tonoplast phospholipids stimulated the activity of purified tonoplast H+-ATPase, but tonoplast glycolipids did not. When tonoplast glycolipids or tonoplast ASG was added in the presence of tonoplast phospholipids, they decreased the phospholipid-induced activation of the tonoplast H+-ATPase; tonoplast GlcCer only caused a small decrease. Steryl glucoside (SG) did not cause any decrease in this activation. Phospholipids, ASG, and GlcCer made up 35 mol%, 20 mol% and 7 mol% of the total lipids of the tonoplast fraction of cultured rice cells, respectively, and these glycolipid levels were enough to depress the phospholipid-induced activation of the tonoplast H+-ATPASE: These results revealed that H+-ATPase activity in the tonoplast may be modulated toward activation and depression by tonoplast phospholipids and glycolipids, respectively. The acylation of SG would be responsible for the depression in the phospholipid-induced H+-ATPase activity. PMID- 11382819 TI - Association of glutathione with flowering in Arabidopsis thaliana. AB - In order to study the relationship between GSH and flowering, wild-type and late flowering mutant, fca-1, of Arabidopsis thaliana were treated with L-buthionine sulfoximine (BSO), a specific inhibitor of GSH biosynthesis, under long-day conditions. BSO treatment of the fca-1 mutant starting at 17 d after imbibition promoted flowering. However, when the treatment was started at 12 d after imbibition, BSO treatment at 10(-4) M resulted in an inhibition of flowering. This inhibitory effect of BSO on flowering was abolished by GSH treatment at 10( 4) M, although GSH treatment at an increased concentration of 10(-3) M clearly delayed flowering. In contrast, BSO treatment of wild-type plants starting at 12 d after imbibition promoted flowering, whose effect was abolished by GSH application. In the fca-1 mutant, whose endogenous GSH levels were high, chilling treatment lowered the GSH levels and promoted flowering, as was the case in the BSO treatment. An A. thaliana mutant, cad2-1, which has a defect in GSH biosynthesis also exhibited late flowering. The late-flowering phenotype of this mutant tended to be strengthened by BSO and abolished by GSH treatment. These results suggest that flowering is associated with the rate of GSH biosynthesis and/or the levels of GSH in A. thaliana. PMID- 11382820 TI - Regulation of the H+ pump activity in the plasma membrane of internally perfused Chara corallina. AB - The role of cytoplasm for the maintenance of the H+ pump activity in Chara corallina internodal cells was examined by the intracellular perfusion technique. Cytoplasm-rich and -poor states were obtained by changing the perfusion time, short-term (less than 2 min) and long-term (more than 5 min), respectively. A large portion of cytoplasm was left by short-term perfusion but most of the cytoplasm was removed by long-term perfusion. The activities of the H+ pump of these two different conditions were examined by measuring current-voltage relation (I-V curve) and conductance-voltage relation (G-V curve) under voltage clamp conditions. The H+ pump conductance decreased to 37%, 9% and zero by short term, long-term and hexokinase perfusion, respectively, whereas the passive channel conductance decreased to 71%, 39% and 73% by short-term, long-term and hexokinase perfusion, respectively. On the other hand, the electromotive-force of the H+ pump (approximately -260 mV) and the passive channel (approximately -130 mV) were not affected by either short- or long-term perfusion. It is indicated that the cytoplasm plays an essential role to regulate the activity of both the H+ pump and the passive channel together with ATP. PMID- 11382821 TI - Magnesium is more efficient than calcium in alleviating aluminum rhizotoxicity in soybean and its ameliorative effect is not explained by the Gouy-Chapman-Stern model. AB - The mechanistic basis for cation amelioration of Al rhizotoxicity in soybean was investigated through a series of studies comparing protective effects of Ca and Mg against Al inhibition of root elongation in a background 0.8 mM CaSO4 solution (pH 4.3). A modified Gouy-Chapman-Stern model was used to evaluate the effect of cations on electrical potential and Al3+ activity at root plasma membrane surfaces. Activities of Al3+ up to 4.6 microM in the background solution inhibited soybean tap root elongation by more than 80%. There was little or no response in root elongation when Ca and Mg were added to background solutions in the absence of AL: When added to Al-toxic solutions in the micromolar concentration range, Mg was 100-fold more effective than Ca in alleviating Al toxicity, whereas both cations were equally effective when added in the millimolar concentration range. The protective effect of micromolar additions of Mg on root elongation was specific for Al and it failed to alleviate La rhizotoxicity. In contrast to wheat, Mg amelioration of Al toxicity to soybean root elongation at low Mg concentration could not be explained by changes in potential and Al3+ activity at the root plasma membrane surfaces as predicted by a Gouy-Chapman-Stern model. These results suggest that Mg is not acting as an indifferent cation when present at low concentration and implies the involvement of a mechanism other than pure electrostatic effects at the root surface. PMID- 11382822 TI - Magnesium ameliorates aluminum rhizotoxicity in soybean by increasing citric acid production and exudation by roots. AB - Superior effectiveness of Mg over Ca in alleviating Al rhizotoxicity cannot be accounted for by predicted changes in plasma membrane Al3+ activity. The influence of Ca and Mg on the production and secretion of citrate and malate, and on Al accumulation by roots was investigated with soybean genotypes Young and PI 416937 which differ in Al tolerance. In the presence of a solution Al3+ activity of 4.6 microM, citrate and malate concentrations of tap root tips of both genotypes increased with additions of either Ca up to 3 mM or Mg up to 50 microM. Citrate efflux rate from roots exposed to Al was only enhanced with Mg additions and exceeded malate efflux rates by as much as 50-fold. Maximum citrate release occurred within 12 h after adding Mg to solution treatments. Adding 50 microM Mg to 0.8 mM CaSO4 solutions containing Al3+ activities up to 4.6 microM increased citrate concentration of tap root tips by 3- to 5-fold and root exudation of citrate by 6- to 9-fold. Plants treated with either 50 microM Mg or 3 mM Ca had similar reductions in Al accumulation at tap root tips, which coincided with the respective ability of these ions to relieve Al rhizotoxicity. Amelioration of Al inhibition of soybean root elongation by low concentrations of Mg in solution involved Mg-stimulated production and efflux of citrate by roots. PMID- 11382823 TI - [Cerebral death: a certitude]. PMID- 11382824 TI - Hemodilution in experimental setting. PMID- 11382825 TI - Haemodilution in clinical anesthesia. PMID- 11382826 TI - [Blood coagulation monitoring during liver transplantation: Sonoclot analysis and laboratory tests]. AB - BACKGROUND: Aim of this paper is to validate blood coagulation data obtained using the Sonoclot thromboelastographic analyser (Sienco Inc., Morrison, USA) by means of standard laboratory tests during orthotopic liver transplantation (OLT). METHODS: DESIGN: comparative study between laboratory data and Sonoclot analysis on simultaneously collected blood samples. SETTING: National Health System Liver Transplantation Center. PATIENTS: fifty-one patients, both males and females, affected by terminal hepatic disease submitted to OLT were enrolled in the study. DATA COLLECTION: simultaneous blood samples were collected during the pre anhepatic, anhepatic and post-reperfusion phases of OLT; coagulation status was assessed by means of either standard Laboratory tests (INR, aPTT, Fibrinogen, PLT, D-Dimer) and Sonoclot analyser data (SonACT, Rate, Time to Peak, Downward Deflection); a statistical analysis was performed (Pearson s chi(2) test). RESULTS: A statistically significative correlation between the analysed data was found. The Sonoclot analyser was useful in identifying platelets dysfunction and was more sensitive to detect fibrinolysis. CONCLUSIONS: The Sonoclot thromboelastographic analyser is a reliable device for monitoring coagulation during OLT. PMID- 11382827 TI - [Anesthesia for lung volume reduction surgery]. AB - BACKGROUND: Lung volume reduction surgery (LVRS) has become a novel palliative procedure for a subgroup of patients with advanced non-bullous emphysema. METHODS: Seventy-six patients with severe emphysema were evaluated: ten patients were considered for lung transplantation and only 24 underwent LVRS. In all patients an epidural catheter was inserted between the T5-T9 space. During one lung ventilation (OLV), ventilatory setting was adjusted to avoid air trapping and/or dynamic hyperinflation and high frequency jet ventilation was used when PaO2/ FiO2 was lower than 60 mmHg in 5 patients. Permissive hypercapnia (PaCO2=53 mmHg) was allowed to avoid hyperinflation and reach hemodynamic stability. RESULTS: During OLV PaO2/FiO2 was 148+/-80 mmHg, PaCO2 53+/-11 mmHg, mPA 27+/-2 mmHg and Qsp/Qt was 38+/-6%. Although the high risk patients, there were no complications due to hypercapnia during surgery. Twenty-three patients were extubated successfully at the end of the surgery (PaO2/FiO2 179+/-34 mmHg and PaCO2 59+/-11 mmHg) and only one patient was not extubated because of air leakage and died for postoperative respiratory failure after 20 days. Another patient died because of sepsis after 15 days. Numeric Ordinal Verbal Scale (by Keele modified) was used for postoperative pain degree at 0, 12th and 24th hours. No patients had pain>2. CONCLUSIONS: In conclusion, a careful anesthesia technique with an accurate intraop monitoring associated with thoracic epidural analgesia even in Video Assisted Thoracic Surgery is suggested in LVRS patients; 12 months postoperative data confirm the validity of the procedure (FEV1 24 AE 36%, FVC 53 AE 70%, RV 265 AE 199% and 6MWT 213 AE 330 m). PMID- 11382828 TI - [The nitric oxide metabolism in the hypoxic, ischemic and reperfused human skeletal muscle cell: clinical and therapeutical observations]. AB - BACKGROUND: The biochemical and metabolic role played by nitric oxide (NO) in course of oxidative stress due to cell hypoxia, ischemia and reperfusion has a determinant relevance in the mitochondrial adaptive changes which antagonize the irreversible morpho-functional damage. In particular conditions, such as in prolonged ischemia and/or exogenous NO supplementation, this element is present in the radical form (NOO*) concurring to peroxidative cell injury. Aim of this study was to investigate these opposite NO aspects in hypoxic, ischemic and reperfused human skeletal muscle tissue. METHODS: Skeletal muscle samples were taken during elective knee orthopedic surgery in 10 consecutive patients. The biopsies were obtained before, after 5+/-1 min and 58+/-2 min from tourniquet application and then after 18+/-3 min following muscle reperfusion. The samples, immediately frozen in liquid nitrogen, were assayed for endocellular free NO following the gas-amperometric method described by Palmerini C. RESULTS: When compared with normoxic tissues, a significant decrease in free NO content was observed in hypoxic samples. After about 60 min of prolonged ischemia the NO levels show an evident increase, while the tissue reperfusion leads to a progressive restoration of physiological content in the cellular free nitric oxide. CONCLUSIONS: The obtained data in hypoxic muscle cell seem to underline the pivotal role played by NO in adapting the cytochrome c oxidase oxidative activity to lower O2 bio-availability. On the other hand the prolonged ischemia leads to a consistent NOO* generation triggered by oxyradical generation and Ca2+ intracellular over load. Even if the tissue reoxygenation restores the normal NO levels it is arguable that the pre-treatment of ischemic cell with antioxidants, Ca-antagonist and Dexamethasone supplementation could represent a crucial and specific therapeutic approach to critically ill patient. PMID- 11382829 TI - Lipid peroxidation, circulating cytokine and endothelin 1 levels in healthy volunteers undergoing hyperbaric oxygenation. AB - BACKGROUND: The aim of the present study was to evaluate the effects of hyperbaric oxygenation on lipid peroxidation, on the release of circulating cytokines (TNFa, IL6, IL1b) and endothelin-1 (ET1). METHODS: EXPERIMENTAL DESIGN: single arm, prospective study. SETTING: ICU hyperbaric division of a University Hospital. PATIENTS: fifteen healthy volunteers (10 male and 5 female, mean age 32+/-7 years) studied during hyperbaric oxygenation divided at random into two groups: group A (7 subjects) and group B (8 subjects). INTERVENTIONS: Both groups were consecutively pressurized at 2 atmospheres (2 atm abs) and 2.8 atm abs, with a constant descending rate of 1 m/min; in accordance with the experimental design, group A breathed pure oxygen continuously through facial masks and group B breathed chamber air during pressurization. MEASURES: Twenty millilitres of blood were drawn from all individuals at the following times: 1) basal, before HBO; 2) after 10 min at 2 atm abs; 3) after 10 min at 2.8 atm abs; 4) 30 min after the end of HBO. In all collected samples thiobarbituric reacting substances were evaluated, using the spectrophotometric technique, IL1 TNF and IL6 serum levels by ELISA and endothelin 1 plasma levels by radioimmunoassay. RESULTS: In both groups, TBARS levels showed a twofold increase (p<0.05) in relation to the baseline, during and after hyperbaric oxygenation. Serum IL6 and IL1b values did not significantly change over the study in any of the volunteers. TNFa amounts significantly increased (p<0.05) during HBO, at 2 atm abs and 2.8 atm abs in both groups, with almost twofold increments. ET1 plasma values increased (p<0.05) in all volunteers during and after HBO: at 2 atm abs (range 7 to 24 pg/ml), 2.8 atm abs (range 7 to 19 pg/ml) and 30 min after (range 8 to 17 pg/ml) in relation to baseline (range 4 to 12 pg/ml). All the studied compounds had a similar trend in the two groups. CONCLUSIONS: Hyperbaric oxygenation in healthy volunteers can induce not only lipid peroxidation, but also liberation of compounds such as TNFa and endothelins, no matter whether pure oxygen is breathed or not. These results suggest that the phenomenon behind this release might be leukocyte activation as induced by HBO. The possible role of ET1 in determining vasoconstriction occurring during HBO is also suggested. PMID- 11382830 TI - Improving postoperative analgesia after axillary brachial plexus anesthesia with 0.75% ropivacaine. A double-blind evaluation of adding clonidine. AB - BACKGROUND: The aim of this prospective, randomized, double-blind study was to evaluate the effects of adding 1 microg/kg clonidine to 20 ml of ropivacaine 0.75% for axillary brachial plexus anesthesia. METHODS: With Ethical Committee approval and written consent, 30 ASA physical status I-II in-patients, undergoing upper extremity orthopedic procedures were randomly allocated to receive axillary brachial plexus block with 20 ml of 0.75% ropivacaine alone (group ropivacaine, n = 15) or 0.75% ropivacaine + 1 microg/kg clonidine (group ropivacaine-clonidine, n = 15). Nerve blocks were placed using a nerve stimulator with the multiple injection technique (stimulation frequency was 2 Hz; stimulation intensity was decreased to < or = 0.5 mA after each muscular twitch; the anesthetic volume was equally divided among arm flexion, arm extension, wrist flexion, and thumb adduction). A blinded observer recorded the time required to achieve surgical block [loss of pinprick sensation in the innervation areas of the hand (C6-C8) with concomitant inability to move the wrist and hand] and first analgesic request. RESULTS: No differences in demography, degree of sedation, peripheral oxygen saturation, and hemodynamic variables were observed between the two groups. Readiness for surgery required 15 min (5-36 min) with 0.75% ropivacaine and 20 min (5-30 min) with the ropivacaine-clonidine mixture. The degree of pain measured at first analgesic request, and consumption of postoperative analgesics were similar in the two groups; while first postoperative analgesic request occurred after 13.8 h (25th-75th percentiles: 9.1-13 h) in the ropivacaine group and 15.2 h (25th-75th percentiles: 10.7-16 h) in the ropivacaine-clonidine group (p = 0.04). CONCLUSIONS: Adding 1 microg/kg clonidine to 20 ml of ropivacaine 0.75% for axillary brachial plexus anesthesia provided a 3 h delay in first analgesic request postoperatively, without clinically relevant effects on the degree of sedation and cardiovascular homeostasis. PMID- 11382831 TI - [Life threatening asthma associated to severe pneumonia and acute myopathy]. AB - A young male was transferred to our intensive care unit (ICU) from the intensive care unit of a local hospital where he was admitted for life-threatening asthma ten days before. As severe hypoxemia, we immediately started inhaled nitric oxide (iNO) that improved significantly pulmonary gas exchange. The first day after admission in our ICU, a chest computed tomography showed a three-lobar pneumonia and, therefore, a broad-spectrum antibiotic therapy was decided. iNO therapy was withdrawn 96 hours after the beginning because of a stable improvement of pulmonary gas exchange and a relative loss of efficacy. Five days after arrival in our ICU, sedative and neuromuscular blocking drugs initiated 15 days before were stopped and, after the awakening, the patient presented tetra-paresis. Muscle biopsy and electromyography indicated an acute myopathy that was probably caused by the association between large doses of steroids and neuromuscular blocking agents. In spite of an intensive physiotherapy program, the patient was extubated only 15 days after admission and he underwent non-invasive mechanical ventilation for further 7 days. The patient was discharged from our ICU 10 days after extubation with a good restore of muscle functioning which was complete two months later. PMID- 11382832 TI - [Coronary artery anomalies and coronary disease: is there a relationship?]. AB - BACKGROUND: It has been suggested that in coronary anomalies the turbulence of the flow might develop atherosclerosis, but this theory remains controversial. The relationship between coronary anomalies and coronary artery disease have been studied. METHODS: Twenty-five patients [15 males, 10 females (mean age 75+/-1.4 years)] with coronary anomalies at coronary angiography were enrolled in a prospective study. Thirteen patients out of 25 (group 1) had critical coronary artery disease (>50% of reduction in luminal diameter). The anomalous vessels passing intramyocardial or between aorta and pulmonary veins were excluded. Twelve patients out of 25 had no sign of atherosclerosis (group 2). At clinical and instrumental follow-up (54+/-8.4 months) the number of cardiovascular events (acute myocardial infarct, unstable angina, silent ischemia, bypass or PTCA interventions, cardiovascular death) was evaluated in the two groups. RESULTS: Age and ejection fraction value were no statistically different in the two groups. Risk factors were significantly (p<0.04) higher in group 1 (smoke, hypertension, hypercholesterolemia, diabetes). The patients number with cardiovascular events was significantly higher in group 1(11 patients vs 2) confirming the independence of the two phenomena. No correlation was found between presence of risk factors and presence of events. A correlation (r=0.70) was found only between previous coronary artery disease and number of events. CONCLUSIONS: No relationship between coronary anomalies and coronary artery disease seems to be confirmed. The real relevance of flow turbulence in the development of atherosclerosis in coronary anomalies needs to be still confirmed. PMID- 11382833 TI - Modifications in cardiovascular functional parameters with aging. AB - BACKGROUND: The constant rise in the number of old people involves a great interest towards the aging of the cardiovascular system. Many authors have studied age-adjusted limits to echocardiographic and functional parameters. The aim of this study is to evaluate some functional parameters so as to define the cardiovascular modifications of the aging heart. METHODS: We have studied 66 patients, 32 of mean age 71+/-4 years (Group 1) and 34 with mean age 33+/-6 years (Group 2), without cardiovascular diseases, through basal echocardiography and exercise test. RESULTS: In comparison to group 2, in group 1 we have observed at rest normal ventricular dimensions with increase of the thickness (SIV 11.2+/-0.4 vs 9+0.6, p=0.01; PP 10.6+/-0.6 vs 9.2+0.4, p=0.01), of the left atrium (41.2+/ 1.08 vs 36.2+1.3, p=0.05), and of the aortic root (33.2+/-1.3 vs, 30.8+0.9, p=0.05), altered diastolic function (E/A 0.7+/-0.14 vs 1.9+/-0.7, p=0.01), with normal systolic indexes. At 100 Watts workload there was an age-related increase in end-diastolic volume (142+/-10.6 vs 127+/-4.3, p=0.01) and an age related decrease in heart rate (HR 100 W 148+/-10.7 vs 169+/-10.9, p=0.05). CONCLUSIONS: In the heart of an elderly subject therefore there are morphological modifications that in the absence of organic pathologies can support the metabolic demands of the organism and adapt to the physiological modifications of the elderly subject. PMID- 11382834 TI - Personal experience of the treatment of ruptured aortic aneurysms. The prognostic evaluation of some parameters. AB - BACKGROUND: The rupture of an aortic aneurysm is the most frequent and most severe complication, with an incidence of approximately 20-40/100,000 persons each year. The aim of this study was to identify the preoperative, intraoperative and postoperative factors that may influence the mortality rate. METHODS: Between January 1996 and December 1999 145 patients underwent emergency abdominal aortic repair surgery. One hundred and twenty-three patients represented a ruptured AAA and 22 a fissured aneurysm. One hundred and thirty-one patients were males and 14 were females; their mean age was 75+/-5.06 years. We selected a number of parameters after a review of the international literature and these were analysed in the two classes of survivors. No statistical analyses were performed on fissured aneurysms owing to the scant number of cases examined. RESULTS: Mortality was 41.4% (44.7% in ruptured aneurysms alone). The following were negative preoperative prognostic factors: old age, hypertension, ASA V, intraoperative anuria and platelet count <100,000. Intraoperative factors included: hypotension, cardiac arrest, onset of anuria, aortic clamping in more than one site, duration of surgery, quantity of RBC and plasma transfused. The main postoperative variables correlated to mortality were: hypotension, hypothermia, onset of anuria, need for further transfusions, high creatinine levels. CONCLUSIONS: Rapid intervention can minimise mortality in structures with expert teams of surgeons and anesthetists experienced in treating this type of pathology. PMID- 11382835 TI - Therapeutic effects of Vascupump treatment patients with Fontaine Stage II B arteriopathy. AB - Intermittent claudication is the primary symptom of chronic obstructive arteriopathies of the lower extremities. Increased walking distance is the aim of all physical or pharmacological treatment of such patients. The study reported aims to assess the increase in distal arterial blood flow in arteriopathic patients achieved by physical treatment using a cuff, generally positioned on the upper third of the thigh to produce pulsed compressions. By tightening for a few milliseconds slightly behind the natural cardiac systole, the cuff reinforces the systolic thrust to increase distal perfusion without making any additional demand, on the cardiac pump. The cuff used is produced by Vascupump Copyright vp El. The trial was conducted on 36 patients with intermittent claudication divided into 3 groups. Group A included 12 patients given Vascupump Copyright treatment only (20 yen 45 minute sessions); Group B included 12 patients given combined treatment with the Vascupump Copyright and endovenous Pentoxillin (3 fl/per diem for 20 days); Group C included 12 patients given Pentoxyfillin alone (3 n per diem for 20 days). Each patient was examined at the start (D0) and end (D20) of treatment as follows: Treadmill tests at 0 and 12% gradients at a speed of 3 km/h. Doppler arterial tensiometry with calculation of the Winsor Index; photoplethysmography. Group A revealed an increase in relative and absolute walking distance of 230% and 224% respectively on the flat and 357% and 343% respectively uphill. Doppler tensiometry revealed a 40.7% improvement in Group A compared to 29% in Group B and 13.6% in Group C. By contrast the improvement in the plethysmographic reading was higher in Group B (+ 119%) than in Group A (+67.3%). These results suggest that the Vascupump Copyright gives better than expected results on Fontaine Stage B arteriopathies. PMID- 11382836 TI - [Gastroesophageal reflux disease and chest pain]. AB - Gastroesophageal reflux disease (GERD) is highly prevalent in the general population. Heartburn and acid regurgitation are considered the typical presentation of GERD, that however might represent a major pathogenic mechanism in patients with chest pain without coronary artery disease. Chest pain in this instance is often successfully treated with high-dose antisecretory drugs, but the pathogenetic mechanism remains unclear. Further research is needed to clarify the role of GERD on the pathogenesis of chest pain and to ascertain whether a medical or surgical anti-secretory treatment could represent a possible approach. Patients with non-cardiac chest pain of unknown origin should be carefully screened for the occurrence of GERD. PMID- 11382837 TI - Small aorta syndrome: hypothesis or reality? AB - A syndrome of peripheral obliterating arterial disease characterised by aortoiliac steno-occlusion is reported in the literature under the name small aorta syndrome, occurring in young women of small stature with relatively typical risk factors. Starting from an analysis of the studies reported in the literature and on the basis of our own results, we have attempted to ascertain whether small aorta syndrome represents an independent nosological entity. By analysing studies on the small aorta syndrome and in the light of a recent study made by our group on arterial diameters measured in cadavers, which highlights a significant correlation between aortic diameter and age, it can be affirmed that a pathology of this nature does not respond to absolute criteria for existence. Therefore, the aortoiliac diameter in women suspected of being affected by small aorta syndrome appears to be broadly in proportion to that expected in healthy women of the same age. Small aorta syndrome does not therefore appear to represent a separate nosological entity. It takes the form of a hypoplastic vascular disorder, which is probably congenital, correlated to other arterial districts in the same subject. However, it may encourage the onset of early symptoms in women of small stature. PMID- 11382838 TI - [Popliteal trauma due to posterior dislocation of the knee]. AB - Popliteal artery injuries have an incidence ranging between 5% and 19% of all traumatic arteria lesions. A high index of amputation, when associated with lesions of the infrapopliteal branches, and a 4-5% overall mortality are recorded in these conditions. Three patients with popliteal artery injury due to posterior knee dislocation were operated upon in our department during the last 12 months. In one case, the physical examination revealed a warm limb, with peripheral pulse; angiography showed a pseudoaneurysm of the popliteal artery with intimal dissection and partial thrombosis. In the last two cases an acute ischemia of the limb was present; in one case the angiographic study showed a complete transection of the artery, while in the second case an obstruction of popliteal artery. In all cases the dislocation was corrected and a the reconstruction carried out by inverted autologus saphenous vein with termino-terminal anastomosis. In all cases a good patency of the popliteal reconstruction was achieved with limb salvage. Prognosis of popliteal injuries is related to an early diagnosis and they should be suspected even in absence of overt signs of acute ischemia. Surgical timing cannot be the same in all cases. The need for a preliminary orthopedic phase with its modality must be established case by case, relating to the severity of ischemia, to the time elapsed between trauma and surgery, to the peculiarities of skeleton and joints lesions. PMID- 11382839 TI - An in vivo method to assess the photostability of UV filters in a sunscreen. AB - An in vivo method was developed in order to study the photostability of UV filters in a sunscreen. This method assesses sunscreen photostability in the emulsion after direct interaction with the skin. For this purpose UV filters were recovered from volunteers' forearms by using the stripping technique, then extracted from tapes and quantified by high-performance liquid chromatography (HPLC). The photostability of the filters tested was evaluated by comparing the amount of filters recovered from the strippings of UV-irradiated skin (40 minimal erythema dose, or MED) versus non-irradiated skin. Sequential analysis of several successive tapes reflected the distribution profile of the filters in the stratum corneum. Photochemical change was observed for one filter: it was shown to undergo a photochemical modification with the appearance of an additional HPLC peak. Moreover, UV filters tested displayed a high affinity for the stratum corneum but presented different distribution profiles. This in vivo method takes into account the interaction of the sunscreen agents with the stratum corneum. Furthermore, unlike spectrometric methods usually used for photostability assessment, it gives quantitative data for each individual filter of a finished product by using an HPLC technique. PMID- 11382840 TI - The ability of electrical measurements to predict skin moisturization. I. Effects of NaCl and glycerin on short-term measurements. AB - Non-invasive methods to evaluate skin hydration by measuring electrical properties are widely used in the cosmetic industry. However, there is still some controversy about factors that affect measurement. For example, concerns have often been expressed about the possible confounding effect of salts, either in the formulation or on the skin. Ionized salts on the skin may increase electrical conductivity and may lead to changes in electrical properties that are not related to increased water content. We have performed a systematic study of the effects of salt, i.e., sodium chloride, and glycerin on the electrical properties of skin as measured by the three most commonly used instruments, the Nova DPM 9003, the Corneometer CM 825, and the Skicon 200. Formulations containing salt from 0-3% and glycerin from 0-10% were tested for their effects at one and two hours after a single application. Salt lowered the readings in the absence of glycerin and increased the reading in the presence of glycerin. For all three instruments, there was a linear correlation between the measurement and the glycerin level in the presence or absence of salt. PMID- 11382841 TI - The ability of electrical measurements to predict skin moisturization. II. Correlation between one-hour measurements and long-term results. AB - We investigated the ability of short-term (one-hour) electrical measurements with three different commonly used instruments to predict the effects of long-term treatment with glycerin-containing formulations on moderately dry leg skin. We report the moisturizing effects of glycerin on healthy female adult skin in a two week study as measured by electrical conductance and capacitance, transepidermal water loss (TEWL), and clinical grading of skin dryness. The test formulations contained 1.5% NaCl and levels of glycerin from 0% to 15%. Results obtained with a smaller cohort of ten subjects, one hour after treatment, were predictive of moisturizing efficacy in the two-week period among twenty subjects with dry leg skin. Our results show that single application tests can be predictive of longer term results with humectant-based moisturizers and that electrical measurements of skin conditions correlate well with skin grades. PMID- 11382842 TI - Kinetics of permeation and metabolism of alpha-tocopherol and alpha-tocopheryl acetate in micro-Yucatan pig sin. AB - The objective of this research was to investigate the permeation and metabolism of alpha-tocopheryl acetate (alpha-TAc) and alpha-tocopherol (alpha-T) from solution and emulsion formulations and to delineate the kinetics of such metabolism. Simple formulations containing alpha-TAc and alpha-T were applied to fresh, viable micro-Yucatan skin dermatomed to a thickness of 250-300 microns, as a finite dose in a flow-through diffusion system. The experiments were stopped at time intervals of 2, 6, 12, and 24 hours. At the end of each time interval, the amounts removed by washing, retained in the stratum corneum (SC), and penetrated into the viable skin and receptor were determined by a validated HPLC method. Receptor concentrations were below the limit of detection. alpha-TAc underwent metabolism in pig skin to the active antioxidant alpha-T. The metabolite appeared as early as two hours after application. The extent of metabolism was highest at 6-12 hours after application. No metabolism was detected in the stratum corneum. Delivery of alpha-T from isopropyl myristate (IPM) solution was more efficient than utilization of alpha-TAc from the same solution. Approximately 1.5% of alpha T yielded the same viable skin concentration as 5% alpha-TAc. Topical application of alpha-tocopherol or its prodrug acetate was capable of enhancing the overall antioxidant capacity of pig skin. The hydrolytic pathway of alpha-TAc leading to the active antioxidant alpha-T could possibly be saturable. PMID- 11382843 TI - Quantitative methods for evaluating optical and frictional properties of cationic polymers. AB - This paper presents three quantitative methods to examine gloss, opacity, and friction of cationic polymers. The adsorption of cationic polymers onto hair and skin can be regarded as a thin film coating. Therefore, optical and frictional properties of polymer films are of significant relevance to the applications of cationic polymers in hair care products. Such properties reflect the desirable hair condition attributes consumers seek in shampoo and conditioner products. Using these test methods, polyquaternium-10 and cationic guar samples of varying molecular weight and cationic substitution were compared. The effect of an anionic surfactant, sodium dodecyl sulfate (SDS), on polymer film properties was also investigated. Neat guar hydroxypropyl trimonium chloride imparts less friction than polyquaternium-10 but dulls the substrate employed in this study. The optical data show that polyquaternium-10 provides greater film clarity and gloss than cationic guars. In the presence of SDS, polyquaternium-10 also displays similar or lower friction than cationic guar. The comparative optical and frictional results are in good agreement with the visual assessment of the cationic polymer films. These results clearly demonstrate that polyquaternium-10 exhibits superior film properties in the forms of both neat polymer and polymer/surfactant complex. In addition, microscopic techniques such as scanning electron microscopy (SEM) and atomic force microscopy (AFM) provide powerful explanations for the differences noted between the two popular classes of cationic polymers. The test methods described in this paper can be utilized to differentiate the upper performance potential of cationic polymers. These objective and standardized test methods derived from the coatings industry are not affected by the variability of hair or the formulation complexity of end products. They can be useful tools in the product development process in quickly screening the relative performance of different polymers. PMID- 11382844 TI - Papers Presented at the 2000 Annual Scientific Meeting (Thursday's Program). PMID- 11382845 TI - Letter from the Editor. PMID- 11382847 TI - Ultraviolet damage on natural gray hair and its photoprotection. AB - The physicochemical properties of natural gray hair obtained from the heads of individuals and as well as commercial samples were investigated. No statistically significant differences were observed in terms of their central maximum diameter, central cross-sectional area, central ellipticity, average tensile strength, and average extent of transverse swelling between gray and black hair. The correlation between the elongation and the contraction of the cross-sectional area of hair fibers during extension was established as a statistically linear function, with a coefficient of 0.758. The damage on natural gray hair from ultraviolet (UV) irradiation were assessed by measuring the following parameters: hair color, Young's modulus, stress-to-break, wet combing force, dynamic advancing contact angle, tryptophan damage, cuticle abrasion, and transverse swelling of hair fiber in 0.1 N NaOH solution. It has been found that gray hair undergoes more severe UV damage and needs more UV protection than dark brown hair. Experimental results indicate that the quaternized UV absorber, cinnamidopropyltrimonium chloride (CATC), delivered from a simple shampoo system, is more substantive on hair and more effective in protecting hair from UV damage than a conventional UV filter. CATC also provided an additional conditioning benefit on hair. PMID- 11382846 TI - Skin optics revisited by in vivo confocal microscopy: melanin and sun exposure. AB - A new confocal prototype dedicated to the exploration of in vivo human skin has been constructed around a laser confocal module (Oz Noran, Inc.) and a skin contact device, assuring perfect stability of skin images. The power of the Argon/Krypton laser source has been limited to 2mW to secure safety, and the laser provides three visible wavelengths: 488, 568, and 647 nm. Optical sections were digitized at video rate, providing easy and rapid measurements of the thickness of epidermal layers and time-resolved information. Unexpected details of the epidermis were recorded with the blue laser line. Melanin provided strong reflection of the basal keratinocytes instead of the absorption expected. The 3D reconstruction of the melanin cap in basal keratinocytes confirmed the behavior of melanosomes acting as myriads of nanomirrors that reflected light. Confocal images of the posterior aspect of the forearm were recorded before sun exposure and then for one month after exposure. There was a 25% increase in the thickness of the stratum corneum. Bright inclusions into the dark nucleus of numerous spinous cells were interpreted as local condensation of chromatin. Numerous bright intercellular filaments were attributed to melanosomes filling up dendrites of melanocytes. A striking observation concerned the lack of melanosome caps in basal keratinocytes. In vivo confocal microscopy affords new insight to the role of melanin and its gradual migration after sun exposure. PMID- 11382848 TI - Examining cationic polysaccharide deposition onto keratin surfaces through biopolymer fluorescent labeling. AB - Fluorescein-labeled polyquaternium-10 and guar hydroxypropyltrimonium chloride were employed to study the deposition behavior of these cationic polymers onto hair from a surfactant system. The influence of the covalently attached fluorescein dye on labeled polyquaternium-10 was examined through rheological studies and comparative studies against data previously obtained from radiolabeled polyquaternium-10. A quantitative method for analyzing the amount of cationic polymer that deposits onto hair during a standard shampooing process has been developed using the labeled cationic polymers. The technique requires digestion of the hair and analysis of the resulting solutions against known standardization curves. It has been found that the molecular weight of the cationic polymers plays the most significant role in influencing the deposition of the polymers from surfactant, a far greater role than either cationic charge or washing cycles. The technique also allows for determination of polymer deposition at various tress locations, allowing for the study of the influence of tress age (i.e., damage) on polymer deposition. The use of fluorescein-labeled polyquaternium-10 also provides a unique opportunity to visualize the deposited polymers on individual hair fibers via fluorescent or confocal microscopy. PMID- 11382849 TI - The use of x-ray fluorescent spectroscopy to study the influence of cationic polymers on silicone oil deposition from shampoo. AB - In this study, x-ray fluorescent spectroscopy was employed, in a non-destructive way, to analyze the influence that water-soluble, cationic hydroxyethylcellulose (i.e., polyquaternium-10) has on the deposition of silicone oil (dimethicone) onto hair. Virgin brown hair tresses were washed with various model shampoos that contained emulsified dimethicone. The shampoos were modified only by the addition or absence of polyquaternium-10. The results indicate that the cationic polymers do influence silicone oil deposition onto hair during the shampooing process. In the absence of cationic polymer, the silicone oils deposit readily, but appear to show "build-up" phenomena upon repeated washings. When a cationic polymer is present in the continuous phase of the shampoo, the build-up phenomena is significantly diminished, and silicone oil deposition remains relatively constant in repeated washings. In addition, we have noted that the molecular weight of the cationic polymer can have a strong effect on silicone oil deposition. It appears that the higher the molecular weight of the polyquaternium-10, the greater the amount of silicone deposition onto the surface of the hair. To demonstrate that the analysis technique has potential applications in commercial shampoos, we examined a commercial "2-in-1" shampoo that contains dimethicone and polyquaternium-10 and found that the data for our simple model shampoos and the commercial shampoo correlated closely. PMID- 11382851 TI - Coalescence of sleep rhythms and their chronology in corticothalamic networks. AB - The cellular substrates of sleep oscillations have recently been investigated by means of multi-site, intracellular and extracellular recordings under anesthesia, and these data have been validated during natural sleep in cats and humans. Although various rhythms occurring during the state of resting sleep (spindle, 7 14 Hz; delta, 1-4 Hz; and slow oscillation, <1 Hz) are conventionally described by using their different frequencies, they are coalesced within complex wave sequences due to the synchronizing power of the cortically generated slow oscillation (main peak around 0.7 Hz). In intracellular recordings from anesthetized animals, the slow oscillation is characterized by a biphasic sequence consisting of a prolonged hyperpolarization and depolarization. Basically similar patterns are observed by means of extracellular discharges and/or field potentials in naturally sleeping animals and humans. The depolarizing component of the slow oscillation is transferred to the thalamus where it contributes to the synchronization of spindles over widespread territories. The association between the depolarizing component of the slow oscillation and the subsequent sequence of spindle waves forms what is termed the K-complex. The slow oscillation also groups cortically generated delta waves. At variance with previous assumptions that the brain lies for the most part in the dark and a global inhibition occurs in resting sleep, cortical cells are quite active in this behavioral state. This unexpectedly rich activity raises the possibility that, during sleep, the brain is occupied to specify/reorganize circuits and to consolidate memory traces acquired during wakefulness. PMID- 11382852 TI - Changes in anti-phosphoserine and anti-phosphothreonine antibody binding during the sleep-waking cycle and after lesions of the locus coeruleus. AB - Cellular responses to many extracellular signals occur through phosphorylation or dephosphorylation of intracellular proteins. To determine whether changes in protein phosphorylation accompany the electrophysiological changes occurring during the sleep-waking cycle, immunocytochemical mapping of cells labeled with anti-phosphoserine and anti-phosphothreonine antibodies was performed on brain sections of sleeping and waking rats. Animals implanted for chronic polysomnographic recordings were sacrificed after either 3h of sleep or 3h of sleep deprivation by gentle handling. Anti-phosphoserine and anti phosphothreonine staining was mainly localized in neurons and was high in some brain regions, such as cerebral cortex and hypothalamus, and low in others, such as the thalamus. In all cases, the number of cells labeled with either antibody in the cerebral cortex was markedly higher in rats sacrificed after 3h of waking than in rats sacrificed after 3h of sleep. Unilateral lesions of the locus coeruleus by local injection of 6-hydroxydopamine were performed in other animals to determine whether the increase in protein phosphorylation during waking was influenced by the activity of the noradrenergic system, which is higher in waking than in sleep. In animals sacrificed after 3h of spontaneous or forced waking, the number of labeled neurons in the cerebral cortex was decreased on the side in which noradrenergic fibers had been lesioned. These results suggest that 1) neurons exist physiologically in different states of phosphorylation, ranging from a state of very high phosphorylation (e.g., in the cerebral cortex) to a state of very low phosphorylation (e.g., in many thalamic nuclei); 2) the fraction of highly phosphorylated neurons in cerebral cortex is higher in waking than in sleep and 3) part of the immunoreactive phosphorylation present in highly labeled cortical neurons is controlled by the locus coeruleus. PMID- 11382853 TI - Decrease in muscarinic M2 receptors from synaptosomes in the pons and hippocampus after REM sleep deprivation in rats. AB - The effects of both REM sleep deprivation and its recovery on pontine and hippocampus muscarinic M2 receptors were investigated in synaptosomes using [3H] AF-DX 384 as a ligand. Animals were divided into three groups: REM sleep deprivation group (small platforms 6.5 cm of diameter); stress group (large platforms 14 cm of diameter) and cage control group. In a second experiment REM sleep-deprived animals were allowed 48 h of recovery. REM sleep-deprived rats showed a reduction in M2 receptors compared with both intact and stress groups. Changes in M2 receptors were also observed after 48 h of recovery from REM sleep deprivation only in hippocampus. The enhancement of acetylcholine release during both REM sleep deprivation and recovery could explain the present findings. PMID- 11382854 TI - Muscle fibre type and obstructive sleep apnea. AB - Muscular pharyngeal structural changes, as fibre type disproportion, have been described in patients affected by Obstructive Sleep Apnea (OSA) and in an animal experimental OSA model. The unsolved question is whether these muscular abnormalities are either secondary to a compensatory increased activity or due to a constitutionally determined reduction of slow-alpha motor neurons. In the present study Medium Pharyngeal Constrictor Muscles (MPCM) of OSA (n = 13) and non-OSA (n = 9) patients have been morphologically evaluated. In addition a needle biopsy of Vastus Lateralis Muscle (VLM) was performed in 5 randomly selected patients of each group. Our results confirmed a specific fibre type disproportion of MPCM of OSA patients compared to non-OSA ones with a type II predominance and aspecific myopathic changes such as fibrosis and central nuclei. No difference was found in the VLM of the two groups. This finding could be explained by a secondary adaptive transformation consequent to nocturnal upper airway resistance in OSA. In fact, it has been demonstrated in human muscle that heavy-resistance training may produce preferential type II fibre hypertrophy in stimulated muscle. PMID- 11382855 TI - c-fos Expression in mesopontine noradrenergic and cholinergic neurons of the cat during carbachol-induced active sleep: a double-labeling study. AB - The interaction of cholinergic and catecholaminergic mechanisms in the mesopontine region has been hypothesized as being critical for the generation and maintenance of active (REM) sleep. To further examine this hypothesis, we sought to determine the pattern of neuronal activation (via c-fos expression) of catecholaminergic and cholinergic neurons in this region during active sleep induced by the pontine microapplication of carbachol (designated as active sleep carbachol). Accordingly, we used two sets of double-labeling techniques; the first to identify tyrosine hydroxylase-containing neurons (putative catecholaminergic cells) which also express the c-fos protein product Fos, and the second to reveal choline acetyltransferase-containing neurons (putative cholinergic cells) which also express Fos. Compared to control cats, active sleep carbachol cats exhibited a significantly greater number of Fos-expressing neurons in the dorsolateral region of the pons, which encompasses the locus coeruleus, the lateral pontine reticular formation, the peribrachial nuclei and the latero dorsal and pedunculo-pontine tegmental nuclei. However, both control and active sleep-carbachol cats exhibited a similar number of catecholaminergic and cholinergic neurons in those regions that expressed Fos (i.e., double-labeled cells). A large number of c-fos-expressing neurons in the active sleep-carbachol cats whose neurotransmitter phenotype was not identified suggests that non catecholaminergic, non-cholinergic neuronal populations in mesopontine regions are involved in the generation and maintenance of active sleep. The lack of increased c-fos expression in catecholaminergic neurons during active sleep carbachol confirms and extends previous data that indicate that these cells are silent during active sleep-carbachol and naturally-occurring active sleep. The finding that cholinergic neurons of the dorsolateral pons were not activated either during wakefulness or active sleep-carbachol raises questions regarding the synaptic mechanisms of activation of these cells during these behavioral states. PMID- 11382856 TI - Pontine injections of nitric oxide synthase inhibitor l-name consolidate episodes of rem sleep in the rat. AB - Dorsal mesopontine cholinergic neurons control rapid eye movement sleep (REMS) and wakefulness and contain nitric oxide (NO) synthase. To assess whether local inhibition of NO synthase has distinct effects on sleep, N-nitro-L-arginine methyl ester, an NO synthesis inhibitor (L-NAME, 80 mM), carbachol, a cholinergic agonist (2, 10 or 50 mM), or saline were microinjected (120-200 nl) into the dorsal mesopontine tegmentum in rats. Sleep-wake cycles were monitored during the subsequent 6 h periods. Compared to control injections, L-NAME changed the pattern of REMS by prolonging individual episodes with a small increase in the percentage time of REMS and no change in slow wave sleep (SWS). Carbachol, at 50 mM, enhanced wakefulness and suppressed both SWS and REMS, especially during the first 2 h post-injection. At the two lower concentrations, carbachol moderately enhanced REMS 2-6 h post-injection by increasing the frequency, rather than duration, of individual episodes. Thus, a reduced NO release in the dorsal pontine tegmentum has a powerful consolidating effect on REMS episodes, whereas the direction of the effect of carbachol on the amount of sleep, and REMS in particular, depends on the magnitude of cholinergic stimulation. The REMS consolidating effects of NO synthase inhibition in the pons may result from modulatory effects of NO on the release of acetylcholine and other neurotransmitters within the dorsal mesopontine tegmentum. PMID- 11382857 TI - Increased dopaminergic transmission mediates the wake-promoting effects of CNS stimulants. AB - Amphetamine-like stimulants are commonly used to treat sleepiness in narcolepsy. These compounds have little effect on rapid eye movement (REM) sleep-related symptoms such as cataplexy, and antidepressants (monoamine uptake inhibitors) are usually required to treat these symptoms. Although amphetamine-like stimulants and antidepressants enhance monoaminergic transmission, these compounds are non selective for each monoamine, and the exact mechanisms mediating how these compounds induce wakefulness and modulate REM sleep are not known. In order to evaluate the relative importance of dopaminergic and noradrenergic transmission in the mediation of these effects, five dopamine (DA) uptake inhibitors (mazindol, GBR-12909, bupropion, nomifensine and amineptine), two norepinephrine (NE) uptake inhibitors (nisoxetine and desipramine), d-amphetamine, and modafinil, a non-amphetamine stimulant, were tested in control and narcoleptic canines. All stimulants and dopaminergic uptake inhibitors were found to dose dependently increase wakefulness in control and narcoleptic animals. The in vivo potencies of DA uptake inhibitors and modafinil on wake significantly correlated with their in vitro affinities to the DA and not the NE transporter. DA uptake inhibitors also moderately reduced REM sleep, but this effect was most likely secondary to slow wave sleep (SWS) suppression, since selective DA uptake inhibitors reduced both REM sleep and SWS proportionally. In contrast, selective NE uptake inhibitors had little effect on wakefulness, but potently reduced REM sleep. These results suggest that presynaptic activation of DA transmission is critical for the pharmacological control of wakefulness, while that of the NE system is critical for REM sleep regulation. Our results also suggest that presynaptic activation of DA transmission is a key pharmacological property mediating the wake-promoting effects of currently available CNS stimulants. PMID- 11382858 TI - Disinhibition of the sleep state-dependent p1 potential in Parkinson's disease improvement after pallidotomy. AB - We previously reported that the P1 or P50 midlatency evoked potential underwent decreased habituation or disinhibition in patients with Parkinson's Disease. This sleep state-dependent response appears to be generated by cholinergic elements of the reticular activating system. We attempted to determine if the decreased habituation or disinhibition of the P1 potential would be altered by bilateral pallidotomy. Twenty-three patients who met inclusion criteria for surgery underwent pre- and post-operative evaluation using a Modified United Parkinson's Disease Rating Scale (UPDRS) and P1 potential recordings. Decreased habituation of the P1 potential was determined using a paired stimulus paradigm in which click stimuli were presented at 250, 500 and 1000 msec interstimulus intervals (ISI). Pre-operatively, patients showed disinhibition of the P1 potential at the 250 msec ISI (60 37% vs. 21 20%) and 500 msec ISI (78 47% vs. 43 31%) compared to age-matched control subjects. Post-operatively, the same patients showed a significant improvement in habituation of the P1 potential at the same ISIs (250 msec 37 21%; 500 msec 43 32%). UPDRS scores for these patients pre-operatively were 59 18 and 24 11 post-operatively, resulting in a significant reduction in symptom severity. We conclude that bilateral pallidotomy resulted in a significant improvement in symptom ratings and reduced the disinhibition of the P1 midlatency evoked response. PMID- 11382859 TI - Obstructive sleep apnea patients use more health care resources ten years prior to diagnosis. AB - Because Obstructive Sleep Apnea Syndrome (OSAS) patients may be treated for comorbidities prior to OSAS diagnosis, we examined the health care utilization records of 181 OSAS patients and those of matched controls. We compared OSA patient health care utilization for a ten-year interval prior to diagnosis to those of randomized age-, gender-, and geographically-matched controls from the general population. We found that OSAS patients used approximately twice the resources (as defined by physician claims and stays in hospital) in the ten years prior to their diagnosis. Physician claims for cases totaled $686,365 ($3,972 per patient) compared to $356,376 ($1,969 per patient) for the controls for the length of the study. Utilization was significantly higher in 7 of 10 years prior to diagnosis. OSAS patients also had more hospitalizations: they had 1,118 nights (6.2 per patient) in hospital versus 676 nights (3.7 per patient) for controls over the ten-year period. Thus OSA patients are heavy users of health care resources ten years prior to diagnosis. PMID- 11382860 TI - Ventilatory responsiveness to progressive asphyxia during sleep in newborn lambs: effects of maternal anemia during pregnancy. AB - Our aim was to determine the effects of intrauterine compromise, induced by maternal anemia, on ventilatory responsiveness of the sleeping newborn to progressive asphyxia. We induced anemia in 6 sheep for the final third of pregnancy and studied their offspring for 2-3 weeks after birth. Lambs from anemic ewes were growth-restricted at birth; they and 6 control lambs were chronically instrumented soon after birth and underwent studies during which we determined ventilatory and arousal responsiveness to a progressive asphyxic stimulus during sleep. During quiet wakefulness, active sleep and quiet sleep, lambs from anemic ewes had elevated end-tidal CO2 levels (FECO2,%) compared to controls. Ventilatory responsiveness (i.e., gradient of relationship between minute ventilation and FECO2) was greater in quiet sleep than in active sleep for both groups of lambs but did not differ between the two groups in either active or quiet sleep. Lambs from anemic ewes had significantly higher FECO2 values than controls before arousing from either active or quiet sleep. Other indices of arousability (time to arousal, percent hemoglobin saturation at arousal) were not different between the two groups. Our results indicate that prenatal exposure to maternal anemia induces fetal growth restriction and elevates the CO2 'set-point' for normal ventilation. It does not, however, produce significant abnormalities in ventilatory responsiveness to progressive asphyxia during sleep. PMID- 11382861 TI - Central administration of a 5-HT2 receptor agonist and antagonist: lack of effect on rapid eye movement sleep and pgo waves. AB - Serotonin (5-HT) has a role in regulating behavioral state and controlling the production of ponto-geniculo-occipital (PGO) waves, though the exact mechanism of action is not known. The most prevailing explanation is that 5-HT exerts its influence on behavioral state and PGO waves by inhibiting and disinhibiting cholinergic cells in the pedunculopontine tegmentum (PPT) and laterodorsal tegmentum (LDT), which have been implicated in their generation. Recent work in rats has demonstrated 5-HT2 receptors on most cholinergic cells in PPT/LDT. We microinfused the relatively specific 5-HT2 agonist, DOI (1-(2,5-dimethoxy-4 iodophenyl)-2-aminopropane), the relatively specific 5-HT2 antagonist, ketanserin, and the nonspecific 5-HT antagonist, methysergide, locally into the peribrachial region of PPT in cats and monitored behavioral state and PGO waves. Neither drug significantly affected behavioral state or PGO wave activity. These results suggest that 5-HT2 receptors associated with cholinergic cells are minimally involved in the control of behavioral state and, together with the recent findings of others, suggest that 5-HT may not modulate PGO wave generation via direct action on cholinergic neurons in PPT/LDT, a departure from the long held but minimally-tested view. PMID- 11382862 TI - Changes in sleep in response to intracerebral injection of insulin-like growth factor-1 (IFG-1) in the rat. AB - Changes in sleep were studied during 6 hours after intracerebroventricular (ICV) administration of Insulin-like growth factor-1 (IGF-1) or the structurally related insulin. IGF-1 was injected either at dark onset (0.05 or 0.5 microgram) or 6 hours after light onset (0.05, 0.5, or 5.0 microgram). The small dose of IGF 1 consistently, albeit modestly, enhanced NREMS over the 6 hour postinjection period in both the dark and the light cycles (REMS increased only at night). The NREMS-promoting activity vanished when the dose was increased to 0.5 microgram, and 5.0 microgram IGF-1 elicited a marked and prompt suppression in NREMS. Heat inactivated IGF-1 (0.05 microgram) did not alter sleep. On a molar base, the NREMS-promoting dose of insulin was higher than that of IGF-1. Late (hours 7-17 postinjection) enhancements in EEG slow wave activity during NREMS were observed after 5.0 microgram IGF-1. The results indicate that IGF-1 can promote NREMS and may contribute to the mediation of the effects of GH on sleep. The acute sleep suppressive activity of the high dose of IGF-1 is attributed to an inhibition of endogenous growth hormone-releasing hormone (GHRH). PMID- 11382863 TI - Sleep and the cholinergic rem sleep induction test in patients with primary alcoholism. AB - Sleep disturbances of alcoholics while actively drinking and at the beginning of, and during, abstinence were frequently reported. Recently, Gillin et al. (1994) showed that a high "REM sleep pressure" at the time of admission to a 1-month inpatient alcohol treatment program predicted the relapse in nondepressed patients with primary alcoholism at 3 months following hospital discharge. We investigated 24 patients with primary alcoholism after 2-3 weeks abstinence in the sleep laboratory; in 15 of these patients the cholinergic REM sleep induction test (CRIT) with 10 mg galanthamine was performed additionally. In comparison with an age- and sex-matched healthy control group, patients had a heightened "REM sleep pressure" including shortened REM latency and increased REM density. A decrease of serotonergic neurotransmission is proposed as being the neurochemical mechanism to explain the results in alcoholic subjects. Follow-up investigations will clarify whether the sleep abnormalities in alcoholism are state- or trait markers and whether they are suitable to predict the relapse risk. PMID- 11382864 TI - The disappearing slow wave activity of hibernators. AB - High and monotonically declining levels of EEG slow wave activity (SWA) occur following arousal from hibernation. Similar profiles of SWA occur in mammals including humans during sleep following periods of prolonged wakefulness, and have been interpreted as reflecting a homeostatic process regulating NREM sleep. It was proposed that even though hibernation appears to be an evolutionary extension of NREM sleep, the low brain temperatures during hibernation are not compatible with sleep restorative processes, and therefore sleep debt accumulates during hibernation and may be a factor triggering periodic arousal. In the present study, golden-mantled ground squirrels were sleep deprived by gentle handling following arousal from hibernation. If the SWA peaks following bouts of hibernation reflect a homeostatic response to an accumulated sleep debt, sleep deprivation should simply displace the SWA which would then occur, and be augmented, during subsequent sleep. In contrast, when animals were sleep deprived following arousal from hibernation, the anticipated SWA peak did not occur during subsequent sleep. It is suggested that the SWA following arousal from hibernation does not represent homeostatic regulation of NREM sleep, but instead some other neurological process involved in the recovery of brain function from an extended period at low temperature. PMID- 11382865 TI - Anatomical demonstration of a medullary enkephalinergic pathway potentially implicated in the oro-facial muscle atonia of paradoxical sleep in the cat. AB - The present study was aimed to compare in detail the distribution within the rostral ventromedial medulla of Methionin-Enkephalin-immunoreactive neurons with efferent projections to the facial or trigeminal motor nuclei, using a double immunostaining technique in colchicine-treated cats. Following cholera toxin B subunit injections in the facial or trigeminal motor nuclei, we found that respectively 55% and 65% of the medium to large-sized retrogradely labeled cells in the lateral part of the nucleus reticularis magnocellularis were Methionin Enkephalin-positive. For both motor nuclei, the double-labeled neurons had similar morphology and size and were located exactly in the same area. They could therefore belong to the same population of reticular enkephalinergic neurons. Based on these and previous anatomical and electrophysiological data, we propose that these enkephalin-containing neurons could participate in the hyperpolarization of brainstem and spinal somatic motoneurons during paradoxical sleep. PMID- 11382866 TI - Prolactin microinjections into the amygdalar central nucleus lead to decreased NREM sleep. AB - Prolactin administered systemically, intracerebroventricularly or locally into the lateral hypothalamus enhances rapid eye movement sleep (REM) when given diurnally and decreases REM when given nocturnally. The amygdala is being recognized as an important modulator of behavioral state, and the central nucleus of the amygdala (CNA) has a high concentration of prolactin fibers and receptors. We microinjected prolactin (10, 100, 250 ng/0.2 microliter saline) or saline alone into CNA of rats and measured the effect on behavioral state. Prolactin produced a dose-dependent decrease in non-REM (NREM), with the effect becoming significant at the high (250 ng) dose. REM was not significantly affected at any dosage. The results indicate a role for prolactin in CNA in the control of NREM. The results are discussed in terms of the amygdala having a broad role in the regulation of behavioral state. PMID- 11382867 TI - Behavioral effects of melatonin treatment in non-human primates. AB - Melatonin treatment has been shown to induce sleepiness and promote sleep in humans. In order to understand the mechanisms by which melatonin acts on human sleep and behavior, it would be useful to have an animal model in which the physiological nocturnal increase in melatonin secretion correlated with nocturnal sleep, i.e., a diurnal species. In this pilot study the oral administration of melatonin to two Pigtail macaques (Macaca Nemestrina) at different times of the day significantly decreased motor activity and promoted earlier sleep onset, as measured actigraphically. The decline in the animals' motor activity occurred within 25-40 min after melatonin ingestion. The duration of motor inhibition was dose dependent. Administration of a 0.05 mg dose induced serum melatonin levels comparable to the peak physiologic concentrations reported in untreated humans and the non-human primates. These data suggest that melatonin may modulate motor activity and sleep pattern in certain diurnally-active primates. PMID- 11382868 TI - Sleep-wake and eeg effects following adenosine a1 agonism and antagonism: similarities and interactions with sleep-wake and eeg effects following a serotonin reuptake inhibitor in rats. AB - Adenosine is currently being investigated as a possible mediator of a homeostatic sleep need. Reports from different laboratories suggest that both adenosine A1 agonists and selective serotonin reuptake inhibitors (SSRI) increase deep slow wave sleep (SWS-2) after an interval. In this study, the sleep-wake effects of the adenosine A1 agonist N6-cyclopentyladenosine (CPA) and the SSRI zimeldine are directly compared in the same animals. Since the SWS-2 increase following SSRIs may be secondary to increased adenosine levels during the initially increased waking, it was also investigated whether the adenosine A1 antagonist 8 cyclopentyltheofylline (CPT) would inhibit the SWS-2 increase following the serotonin reuptake inhibitor. Both the adenosine A1 agonist CPA and the SSRI zimeldine increased SWS-2 after an interval. Both drugs increased slow wave activity and decreased 9-20 Hz activity during SWS-2. Both the adenosine A1 antagonist CPT, zimeldine and the two drugs combined initially increased waking and subsequently increased SWS-2 after 2 or 4 h. All treatments increased 2-6 Hz activity in SWS-2 after 2h. Thus, CPT did not antagonize the SWS-2 increase of zimeldine. Based on the sleep and power spectral effects it is suggested that the adenosine A1 antagonist potentiated the zimeldine effect, possibly due to antagonism of adenosine A1 inhibition of serotonin release. The data indicate that the delayed SWS-2 and slow wave activity increases following zimeldine are not due to increased stimulation of adenosine A1 receptors following the initial sleep loss. PMID- 11382869 TI - Abolition of the neocortically monitored theta rhythm after ibotenic acid lesion of the parafascicular nucleus in behaving rats. AB - Seven adult Sprague-Dawley rats, chronically implanted with standard electrodes to monitor frontoparietal electroencephalographic (EEG) and nuchal electromyographic (EMG) activity, received, under deep anesthesia, unilateral or bilateral microinjections of ibotenic acid in the lateral part of the parafascicular nucleus of the thalamus. Four days after the injections (corresponding to the period of neuronal destruction), obliteration of the oscillatory activity in the theta range was found on the side ipsilateral to the injection, while on the intact hemisphere the rhythm was well developed. The asymmetry between the two hemispheres was particularly evident during REM sleep but was also seen during attentive but immobile alertness. In bilaterally injected rats, the neocortical theta rhythm was abolished on both hemispheres. These results suggest that in freely-moving rats the lateral parafasciculus neurons are part of the network on which the emergence of the theta rhythm relies. PMID- 11382870 TI - Differential responses of brain stem neurons during spontaneous and stimulation induced desynchronization of the cortical eeg in freely moving cats. AB - The EEG is desynchronized during wakefulness and REM sleep. There are awake and REM sleep-related neurons in the brain stem. This study was carried out to investigate if the same neuron in the brain stem reticular formation may be responsible for EEG desynchronization during wakefulness and REM sleep. Single neuronal activity was recorded in chronically prepared freely moving normal cats and their activities were correlated with EEG desynchronization during spontaneous wakefulness, REM sleep, and during wakefulness induced by stimulation of the brain stem reticular formation. A majority of the neurons showed an increased firing associated with spontaneous EEG desynchronization during wakefulness and REM sleep, however, about 55% of them showed a similar behavior during stimulation-induced desynchronization. It was found that responses of a majority of the neurons during stimulation-induced desynchronization were similar to that of their firing rate during EEG desynchronization associated with spontaneous wakefulness irrespective of their behavior during REM sleep; the REM ON neurons were not affected by the stimulation-induced desynchronization. A majority of the neurons which showed an increased firing during spontaneous and stimulation-induced EEG desynchronization received an excitatory input from the brain stem reticular formation. The results of this study suggest that although some neurons may be common, there is a strong possibility that the same neuron in the brain stem reticular formation is not involved in EEG desynchronization during wakefulness and REM sleep. PMID- 11382871 TI - Sleep disturbances and eeg slowing in alzheimer's disease. AB - Changes in sleep structure, and especially REM sleep, and in EEG activation were studied in relation to the cholinergic deficit found in Alzheimer's Disease (AD). With respect to sleep architecture, only REM sleep percent was reduced in AD patients compared to controls as a result of a decrease in mean REM episode duration. Different results were obtained in patients with progressive supranuclear palsy (PSP). These results are discussed with respect to the role of brainstem and forebrain cholinergic populations in REM sleep generation in humans. More importantly, it was shown by means of spectral analyses that EEG slowing is much more prominent in REM sleep than in wakefulness in AD. Furthermore, there is a distinct topographical pattern of REM sleep EEG slowing in AD patients which is in agreement with findings from neuroradiological and neuropathological studies. Using the ratio of slow over fast frequencies from the temporal regions, a correct classification of 90.4% of subjects was obtained for the REM sleep EEG. This discrimination rate is the best marker of AD so far using a single measure. Quantitative REM sleep EEG was also used to evaluate patients' biological response to cholinergic treatments. Finally, we present here preliminary data on the progression of EEG slowing in wakefulness and in REM sleep. After six months on a placebo, there was only a decrease in alpha activity in wakefulness over all regions studied. No changes were observed for REM sleep. PMID- 11382872 TI - Effects of gamma-hydroxybutyrate on ventral tegmental unit activity in the rat: considerations on rem sleep control. AB - The effect of gamma-hydroxybutyrate (GHB) administration on spontaneously active dopaminergic cells of the ventral tegmental area (VTA) was determined using extracellular single unit recordings in urethane-anesthetized rats. High doses (160-250 mg/kg, i.p.) of GHB reversibly decreased firing rate in 63.6% of the cells tested (n=11); remaining cells (36.4%) were unaffected. When the GHB receptor antagonist NCS-382 (10 mg/kg, i.p.) was co-administered with GHB at high doses, 50% of the cells became excited while remaining cells were unaffected. Of the 34 cells tested with GHB at low doses (10 mg/kg, i.p.), 21 (61.8%) changed their firing activity. Of these, 12 (57.1%) were excited, five (23.8%) were inhibited, and four (19.0%) were first excited then totally inhibited (E/Ipattern). Out of the three E/I cells tested, two resumed their firing activity after apomorphine (50 microgram/kg s.c.), showing that they were in a state of depolarization inactivation. When NCS-382 (10 mg/kg, i.p.) was co administered with GHB at low doses, only two of the seven cells tested (28.6%) changed their firing activity, both with excitations. We conclude that only low doses of GHB selectively activate GHB receptors. Mechanisms by which low doses of GHB facilitate REM sleep are discussed. PMID- 11382873 TI - Impaired circadian waking arousal in narcolepsy-cataplexy. AB - The 24-hour sleep/wake distributions of untreated patients with narcolepsy cataplexy and matched normal habitual nappers were compared using home ambulatory monitoring. Subjects followed their usual sleep patterns including, for the habitual nappers, a self-selected daytime nap. There were no differences in 24 hour totals of sleep between groups other than a small increase in SWS in narcolepsy. Narcolepsy showed greater amounts of day sleep (stages 2, SWS, REM and total sleep) and less night sleep (stage 2, total sleep). Data were collapsed into 5 min epochs and entered into a matrix. The data in the two groups were then "wrapped" (re-aligned) around the 24 hours with phase 0 as each of the times of: evening sleep onset, onset of SWS, mid-point of night sleep and moment of morning awakening. In habitual nappers alignment beginning at morning wake-up produced the highest amplitude, least temporal dispersion and greatest kurtosis of daytime sleep (naps). The 24-hour sleep/wake distribution curves of both subject groups (data aligned at morning wake-up) based on collapsed data into 5 min bins then underwent curve fitting using 15th order polynomial regression. As with visual analyses of the raw data, the curve fits confirmed that the peak in daytime sleep propensity in narcoleptics was earlier by about 40 (2.66 hours). It was concluded that decreased daytime amplitude of a circadian arousal system was the most parsimonious explanation for the increased amount, broader temporal distribution and relative phase advance of day sleep in narcolepsy and that, as well, such a mechanism could explain a number of other features of the disease. PMID- 11382874 TI - SCN controlled circadian arousal and the afternoon "nap zone". AB - This paper outlines a conceptual model for the regulation of the circasemidian sleep propensity process with emphasis on a possible mechanism of the afternoon "nap zone". It is proposed that the afternoon nap zone is due to increasing sleep propensity after morning wakening (Borb ly's Process-S) being overwhelmed by a light-sensitive SCN-dependent circadian arousal process of the type discovered by Edgar et al., (1993) and currently being identified in its pathways and neurochemistry by Jouvet and colleagues. It is maintained that this arousal process is reflected in the circadian core body temperature pattern, and that under normal entrained conditions the latter does not resemble a sine-wave or skewed sine-wave. Rather it is very asymmetrical in time and somewhat asymmetrical in amplitude. Cosinor type analyses which enforce symmetry in time and amplitude are therefore ill suited to adequately curve-fit the empirical data. The shape of the circadian arousal system was clarified by meta-analyses of data from three laboratories for three conditions: the normal entrained state, the constant routine, and temporal isolation. Under normal entrained conditions for about one-third of the circadian day core body temperature, and therefore the assumed intensity of the circadian arousal system, is below the mesor with the nadir being at about 0500h; and for about two-thirds of the circadian day it is above the mesor with the acrophase on average being at about 2100h. For modeling purposes, the homeostatic process (Process-S) employed the actual data of the Zurich laboratories for night sleep, but altered the equation for the daytime period to ensure an exponential increase after wake-up. Combining these modified processes indicated that the nap zone could be explained, as predicted, by an increasing homeostatic pressure for sleep across the daytime being reversed by the circadian arousal process. This 2-process combination predicted quite well the shape of the entire circasemidian sleep/wake propensity process and can explain the presence of morning sleep inertia without requiring a third process. It would appear that the circadian arousal process can be modified in phase and in amplitude by a number of normal and pathological conditions. PMID- 11382875 TI - Injection of 6-hydroxydopamine into the ventral tegmental area suppresses the increase in arterial pressure during REM sleep in the rat. AB - We have examined the effect of injection of 6-hydroxydopamine (6-OHDA) into the ventral tegmental area (VTA) on the changes in arterial blood pressure (AP) and heart rate (HR) during the transition from non-rapid eye movement (NREM) sleep to REM sleep. The 6-OHDA-treated rats showed suppression of the increase of AP and HR during REM sleep and of theta frequency in the cortical electroencephalogram (EEG) during wakefulness (W) and REM sleep. It is suggested that midbrain dopaminergic neurons are involved in the control of AP and HR during REM sleep and in the EEG theta activity. PMID- 11382876 TI - Cervical positional effects on snoring and apneas. AB - We examined the effects of cervical position on the Obstructive Sleep Apnea Syndrome (OSAS) through the use of a custom-designed cervical pillow which promoted neck extension. Twelve subjects with OSAS were recruited from a tertiary sleep disorder clinic population. Of the twelve subjects, three had mild cases of OSAS, four had moderate cases, and the remaining five had severe cases. The subjects used their usual pillows during two consecutive recorded baseline nights in our laboratory. The subjects then used the cervical pillow for five days at home, and returned for two consecutive recorded nights at our laboratory while using the cervical pillow. During the nights in our laboratory, the subjects completed questionnaires, were videotaped to record head and body position, and had their breathing parameters recorded during sleep. Subjects with mild OSAS cases had a non-significant improvement in the severity of their snoring and a significant improvement in their respiratory disturbance index with the cervical pillow, while subjects with moderate OSAS cases showed no improvement in these parameters. Subjects with severe OSAS cases showed slight improvement in some measures of their abnormal respiratory events during the experimental period. PMID- 11382877 TI - From snoring to sleep apnea in a Singapore population. AB - We have no information on snoring and obstructive sleep apnea (OSA) in our population, which is predominantly Chinese. Our perception is that sleep apnea syndrome is more common than the 2-4% prevalence (Young et al., 1993) often quoted, judging from the experience in our sleep disorder unit. We studied the snorers in an adult population in Singapore and then went on to see how many snorers suffer pathological apnea and sleep apnea syndrome. Room partners, 220 of them aged 30-60 years, were interviewed for their observation of snoring among each other. 106 consecutive habitual loud snorers of a similar age group in the same population were studied with polysomnography in our sleep laboratory. An apnea index greater than 5 was considered pathological. 24.09% were loud habitual snorers. 87.5% of loud habitual snorers had significant obstructive apneas on the polysomnogram and 72% of these apneics complained of excessive daytime sleepiness (EDS). Given the clinical observation that all apneics snored, by extrapolating these figures, we guess that sleep apnea syndrome affects about 15% of the population. Multiple Sleep Latency Tests validated EDS in our cases with clinical hypersomnia. Hypersomnolence was significantly related to the poor delta wave sleep. Contrary to what was believed, OSA occurred predominantly in stage 1 and 2 non-rapid eye movement (NREM) sleep rather than in REM sleep. The frequent arousals prevented sleep going beyond stage 1 and 2. PMID- 11382878 TI - Selective slow-wave sleep (SWS) deprivation and SWS rebound: do we need a fixed SWS amount per night? AB - Visually scored delta activity (stages 3 and 4, SWS) as well as computerized delta activity measures increase after total and selective sleep deprivation. It is, however, still controversial if SWS amount is only a function of prior waking duration, or if it is related to the structure of the previous sleep period (i.e., to the time spent in SWS). In order to clarify if the amount of SWS is crucial in determining SWS recovery, we selectively deprived SWS during two nights to assess the presence of a compensatory SWS rebound in the following recovery night. Ten normal males slept for 6 consecutive nights in the laboratory. After an adaptation and two baseline nights (BSL; BSL-A), selective SWS deprivation was accomplished for two consecutive nights (DEP-1; DEP-2), by means of an acoustic stimulation technique. A recovery (REC) night then followed. An almost complete selective SWS suppression during both deprivation nights was achieved. A significant increase of S4 and SWS in the REC as compared to the BSL A paralleled a significant shortening of S3 and S4 latencies. S2 percentage significantly increased during both DEP nights with respect to the other experimental nights. There was no significant difference among nights with regard to total sleep time, percentage of REM sleep, stage 1, movement time, number of awakenings and number of movement arousals, indicating that the acoustic stimulation technique did not dramatically disrupt normal sleep continuity and architecture. These results indicate that SWS rebound after selective SWS deprivation can be ascribed to the loss of SWS accumulated during two consecutive nights, further supporting the idea that the delta sleep amount is more linked to SWS in the previous sleep periods than to the total sleep duration. PMID- 11382879 TI - Narcolepsy-cataplexy syndrome associated with DRB1*0806-DQB*0602 haplotype in a Caucasian patient. AB - Narcolepsy-Cataplexy (NC) is a neurological disorder associated with the human leukocyte antigen HLA DR2. This is a prerequisite for the disease in 95 to 98% of Caucasian patients. It has been demonstrated that the HLA DQB1*0602 allele is a better marker for narcolepsy than DRB1*1501 (DR2). We present a DR-negative and DQB1*0602-positive Caucasian Spanish patient with a very unusual genotype. A 20 year-old male presented with a 12-year history of excessive daytime sleepiness and sudden muscle weakness caused by laughter and disturbed nocturnal sleep. He had never presented hypnagogic hallucinations or sleep paralysis. The family history was negative. Physical and neurological examinations were normal. The Epworth Sleepiness Scale score was 21/24, The Ullanlinna Scale score was 20/40. The polysomnographic recording showed short sleep latency, increased percentage of stage 1 (St 1), increased number of body movements and decreased sleep efficiency index. MSLT data: mean sleep latency of 1 minute and three sleep onset rapid eye movement (REM) periods (SOREMPs). HLA phenotype: A1, A11; Cw5, Cw7; B44, B39; Bw4, Bw6; DR4, DR8; DR53; DQ6, DQ8 and at the gene level: DRB1*0402, DQB1*0302; DRB1*0806, DQB1*0602. The DRB1*0806 and DQB1*0602 genotype is very infrequent in NC and identical to one African-American case in the series by Mignot et al. (1997a), and to a Caucasian case in another series by Mignot et al. (1997b). This indicates the genetic heterogeneity of the NC. PMID- 11382880 TI - Correlation between blood adenosine metabolism and sleep in humans. AB - Blood adenosine metabolism, including metabolites and metabolizing enzymes, was studied during the sleep period in human volunteers. Searching for significant correlations among biochemical parameters found: adenosine with state 1 of slow wave sleep (SWS); activity of 5'-nucleotidase with state 2 of SWS; inosine and AMP with state 3-4 of SWS; and activity of 5'-nucleotidase and lactate with REM sleep. The correlations were detected in all of the subjects that presented normal hypnograms, but not in those who had fragmented sleep the night of the experiment. The data demonstrate that it is possible to obtain information of complex brain operations such as sleep by measuring biochemical parameters in blood. The results strengthen the notion of a role played by adenosine, its metabolites and metabolizing enzymes, during each of the stages that constitute the sleep process in humans. PMID- 11382881 TI - A GABAergic pontine reticular system is involved in the control of wakefulness and sleep. AB - The present work is the first in a series of studies designed to examine the role of a brainstem GABAergic system in the control of the behavioral states of sleep and wakefulness. GABA, muscimol (a GABAA receptor agonist) and bicuculline methiodide (a GABAA receptor antagonist) were microinjected, separately, into the nucleus pontis oralis (NPO) in three chronic, unanesthetized cats. The effects of these microinjections on the behavioral states of sleep and wakefulness were then examined. The injection of either GABA or muscimol induced wakefulness; quiet sleep and active sleep were suppressed. In contrast, the injection of bicucculline induced a prolonged state that was similar to naturally-occurring active sleep. These findings indicate the existence of GABAergic processes capable of controlling the activity of neurons within the NPO that are involved in the control of sleep and waking states. Specifically, these data suggest that cells within the NPO must be tonically inhibited by a GABAergic brainstem system in order for the state of wakefulness to be generated and maintained. PMID- 11382882 TI - Alterations in monoamine neurotransmitters and dendritic spine densities at the medial preoptic area after sleep deprivation. AB - The experiments were conducted on 24 adult male Wistar rats to find out the alterations in the levels of monoamines and dendritic spine densities in the medial preoptic area and cortex after total sleep deprivation. Noradrenaline was reduced in the medial preoptic area, though there was no significant change in the cortex. Dopamine and serotonin were decreased both in the medial preoptic area and in the cortex. Dendritic spine counts in the medial preoptic area and the motor cortex were increased after total sleep deprivation. Enhanced release of the monoamines and their subsequent breakdown during sleep deprivation could be responsible for the decreased levels of the transmitters. An increase in synaptic activity, resulting in the enhanced release of the transmitters, might be responsible for the increased spine density after total sleep deprivation. Localized changes in noradrenaline levels at the medial preoptic area suggest its involvement in sleep genesis and maintenance, though its possible contribution to other functions like thermoregulation and reproduction cannot be ruled out. As the available literature does not indicate a role for serotonin and dopamine at the medial preoptic area in sleep regulation, these changes may represent their participation in non-sleep functions. PMID- 11382883 TI - Are there non-monoaminergic paradoxical sleep-off neurons in the brainstem? AB - Using extracellular single unit recording in the medulla of freely moving cats, we have found a population of PS-off ("Type II") neurons that are distinct from the classically described monoaminergic PS-off ("Type I") neurons. The presumed non-monoaminergic Type II PS-off neurons (n=22) showed a relatively high rate of tonic discharge during both quiet waking and slow-wave sleep (10.4+/-4.1 and 9.3+/-3.1 spikes/sec, mean +/- S.D., respectively) and a marked overall decrease in discharge rate during PS (0.3+/-0.4 spikes/sec). In contrast to the presumed monoaminergic PS-off neurons (n=62), Type II PS-off neurons showed short-lasting phasic discharges during PS, often in association with rapid eye movement and PGO wave bursts. These Type II neurons were all characterized by a short action potential which was significantly different from that of the monoaminergic PS-off neurons described so far. Five out of 22 neurons were identified antidromically by stimulation of the ventrolateral reticulospinal tract (vlRST) at the caudal medulla, while 2 of the 22 were identified antidromically by stimulation of the peri-locus coeruleus alpha of the mediodorsal pontine tegmentum. Their mean conduction velocity (7.2+/-1.9 m/sec) was significantly higher than that (0.9+/ 0.3 m/sec) of the presumed monoaminergic PS-off neurons which were identified exclusively by stimulation of the vlRST. In addition, when examined during the sleep-waking cycle, the antidromic responses of Type II PS-off neurons were either completely blocked or reduced, with a prolongation of antidromic latency during PS. Most of these neurons were located in medullary structures containing no, or virtually no, monoaminergic neurons, and none responded by inhibition to systemic administration of serotonergic or adrenergic autoreceptor agonists. These findings indicate the existence, in the medulla, of non-monoaminergic PS off neurons that would play an important role in PS generation. PMID- 11382884 TI - Frontal predominance of a relative increase in sleep delta and theta EEG activity after sleep loss in humans. AB - The effect of sleep deprivation (40 h) on topographic and temporal aspects of electroencephalographic (EEG) activity during sleep was investigated by all night spectral analysis in six young volunteers. The sleep-deprivation-induced increase of EEG power density in the delta and theta frequencies (1-7 Hz) during nonREM sleep, assessed along the antero-posterior axis (midline: Fz, Cz, Pz, Oz), was significantly larger in the more frontal derivations (Fz, Cz) than in the more parietal derivations (Pz, Oz). This frequency-specific frontal predominance was already present in the first 30 min of recovery sleep, and dissipated in the course of the 8-h sleep episode. The data demonstrate that the enhancement of slow wave EEG activity during sleep following extended wakefulness is most pronounced in frontal cortical areas. PMID- 11382885 TI - Sexual behavior in sleep, sleepwalking and possible REM behavior disorder: a case report. AB - Seven cases of sexual behavior during sleep (SBS) have been recently reported. The subjects had histories of behavioral parasomnias as well as positive family histories of parasomnia. A 27 year-old man with a history of sexual behavior during sleep was reported. His sleep history disclosed sleepwalking (SW) since 9 years of age. He also developed episodes of highly disruptive and violent nocturnal behavior with dream enactment at age 20 years, which often resulted in physical injuries either to himself or his wife and infant. His wife also reported episodes of amnestic sexual behavior that began 4 years before referral. During the episodes, the patient typically procured his wife, achieving complete sexual intercourse with total amnesia. Physical and neurological diagnostic workups were unremarkable. Family history disclosed sleepwalking in his brother. He was put on 2mg/day of bedtime clonazepam with a remarkable clinical improvement. This case involves either the combination of violent and non-violent sleepwalking with SBS, or the superimposition of presumed REM sleep behavior disorder (RBD) on top of preexisting SW in a man who also developed SBS in adulthood. Thus, this is a case report of probable parasomnia overlap syndrome. PMID- 11382886 TI - Sleep stages preceding spontaneous awakenings in the elderly. AB - Spontaneous awakenings from sleep were studied in a group of 21 elderly subjects (mean age 69.29+/-3.02 years) free of neurological and somatic diseases. The prevalence of awakenings and the duration of waking bouts were analyzed with regard to the prior sleep state. The results showed an increased frequency of awakenings during Stage 2 NREM in the elderly, who wake out of Stage 2 NREM no less frequently than out of REM sleep. This trend is different from that observed in younger subjects (babies and young adults), where a clear prevalence of REM sleep awakenings has been reported. The duration of Stage 2 interrupted by awakening was shorter than Stage 2 followed by sleep. The duration of waking bouts did not differ according to the preceding sleep stage. It is suggested that the relative inability to sustain Stage 2 may be a mechanism which contributes to the difficulty of sleep maintenance in the elderly. PMID- 11382887 TI - Brainstem afferents of the cholinoceptive pontine wave generation sites in the rat. AB - The present study was designed to investigate the distribution of brainstem neurons projecting to the pontine wave (P-wave)-generating sites in the rat. In six rats, biotinylated dextran amine (BDA) was microinjected into the physiologically identified cholinoceptive P-wave generation site. In all cases, microinjections of BDA in the cholinoceptive P-wave generating site resulted in retrograde labeling of cell bodies in many parts of the brainstem. The majority of those retrogradely labeled cells were in the pedunculopontine tegmentum, pontine reticular nucleus oralis, parabrachial nucleus, vestibular nucleus, and gigantocellular reticular nucleus. The results presented in this study provide anatomical evidence that the cholinoceptive P-wave generation site in the rat receives anatomical projections from other parts of the brainstem known to be involved in the REM sleep-generation mechanism. PMID- 11382888 TI - REM sleep in response to light and dark in congenic albino and pigmented F344 rats. AB - Sleep was recorded in congenic F344 albino (c/c) and pigmented (c/+) rats while they were exposed to various light-dark schedules at 10, 50 and 100 lux. In short LD schedules (1:1 and 3:3), both c/c and c/+ rats had similar patterns of NREM and waking in the light and dark. NREM was higher in the light and there was more wakefulness in the dark. These differences were accentuated with increased light intensity. In contrast, substantial effects on REM sleep were seen only in the c/c rats and increased light levels also enhanced these effects. REM sleep in pigmented c/+ rats was virtually unaffected by lighting changes. These results indicate that different systems are involved in regulating sleep-waking and REM sleep responses to light and further that these systems are differentially affected by alleles at (or near) the c locus and/or albinism. PMID- 11382889 TI - Sleep spindles and arousal: the effects of age and sensory stimulation. AB - This study assessed a proposed sleep-preserving role for sleep spindles by evaluating variations in this activity as a function of factors, both naturally occurring and experimentally induced, known to affect and effect arousal from sleep. These factors included age, auditory stimulation, and experimentally induced arousal from sleep. Analyses were based on data from 84 males (5-49 yrs. old) from normal and clinical (hyperactive, enuretic, and chronic pain) populations who had participated in sleep auditory arousal threshold studies involving adaptation and 1-2 experimental nights. Spindles on experimental nights were visually analyzed and incidence determined for the two minutes preceding and throughout all Stage 2 arousal attempts. Prestimulation spindle occurrence in 39 preadolescent subjects with two experimental nights did not vary significantly from night-to-night, and prestimulation period comparisons between clinical groups and their respective controls were also non-significant. Anticipated relationships between spindle activity and indices of arousal-either inverse with respect to known variations in arousal threshold, i.e., decreases with age and across the night, or direct with respect to stimulus intensity particularly on trials when arousal did not occur-were not observed. Instead, all age groups showed significant decreases in spindle density as an increasing negative function of stimulus intensity. These findings suggest that to the extent to which sleep spindles can be considered to play a role in sleep preservation by inhibiting or attenuating potentially arousing stimuli, these effects appear to be restricted to endogenously generated stimuli and are passive rather than reactive in nature. PMID- 11382890 TI - Chewing can relieve sleepiness in a night of sleep deprivation. AB - Night sleepiness in two groups of student volunteers who stayed awake for one night was assessed at standardized time (22:00, 01:00, 04:00, 07:00) by the Stanford Sleepiness Scale (SSS). One group (N=21) chewed the chewing gum from midnight until the end of the experiment in the morning, while the other group (N=43) was not chewing at all. The results show that both groups at the initial assessment at 22:00 were not sleepy, with similar SSS scores. Sleepiness in both groups appeared after midnight, worsening towards the morning. The students who were chewing from midnight assessed their sleepiness as lower than the students who were not chewing, which was more marked at 01:00 and 04:00. In the group of medical professionals, nurses and technicians, sleepiness was assessed by SSS in a routine night shift when they, according to their own experience, had the most difficulty overcoming it. Immediately after the assessment they chewed the chewing gum (N=60) or stood/walked (N=27) for 15 minutes. At the end of the fifteenth minute, they assessed their sleepiness again. After 15 minutes of treatment both groups of medical professionals assessed their sleepiness as relieved, with a lower SSS score, more markedly in the chewing group. The obtained results seem to indicate that chewing may alleviate sleepiness in professionals and nonprofessionals who stayed awake through the night. PMID- 11382891 TI - Effects of 48 hours sleep deprivation on human immune profile. AB - It is a common belief that sleep deprivation increases the susceptibility to diseases. In order to evaluate the effects of sleep deprivation on immune profile in humans, peripheral venous blood was obtained from sixteen healthy young male volunteers. Ten of the volunteers underwent 48 hours of sleep deprivation and the other six maintained their regular sleep schedule and acted as controls. The first blood samples were taken at the end of the first polysomnographic recording at 8:00 a.m. After this sampling, ten subjects were sleep deprived for 48 hours in sedentary conditions. The second and third blood samples were taken at the 24th and 48th hours. The subjects were recorded again to verify rebound effects of sleep deprivation after the third blood sampling. In this second polysomnographic recording, all sleep-deprived subjects showed slow wave and REM sleep rebound. The last blood samples were taken at the 72nd hour of study at 8:00 a.m. CD4, CD8, CD5, CD16, CD19 surface antigen positive lymphocyte subsets, serum IgG, IgM, and cortisol levels were assessed in all samples. Our results showed that the proportion of NK cells were decreased during sleep deprivation and returned to normal values after recovery sleep. In the control group, we did not observe any changes in the same direction as the sleep-deprived group. PMID- 11382892 TI - REM sleep enhancement and behavioral cataplexy following orexin (hypocretin)-II receptor antisense perfusion in the pontine reticular formation. AB - Orexin (hypocretin)-containing neurons of the hypothalamus project to brainstem sites that are involved in the neural control of REM sleep, including the locus coeruleus, the dorsal raphe nucleus, the cholinergic zone of the mesopontine tegmentum, and the pontine reticular formation (PRF). Orexin knockout mice exhibit narcolepsy/cataplexy, and a mutant and defective gene for the orexin type II receptor is present in dogs with an inherited form of narcolepsy/cataplexy. However, the physiological systems mediating these effects have not been described. We reasoned that, since the effector neurons for the majority of REM sleep signs, including muscle atonia, were located in the PRF, this region was likely implicated in the production of these orexin-related abnormalities. To test this possibility, we used microdialysis perfusion of orexin type II receptor antisense in the PRF of rats. Ten to 24 hours after antisense perfusion, REM sleep increased two- to three-fold during both the light period (quiescent phase) and the dark period (active phase), and infrared video showed episodes of behavioral cataplexy. Moreover, preliminary data indicated no REM-related effects following perfusion with nonsense DNA, or when perfusion sites were outside the PRF. More work is needed to provide precise localization of the most effective site of orexin-induced inhibition of REM sleep phenomena. PMID- 11382893 TI - Letter to the editor: hypothesis for the neurophysiology of dreaming. AB - During wakefulness, the cerebral cortex, which is responsible for generating mental activities, is activated by brain stem ascending influences. This is evidenced by classic electrophysiological field and unitary activities, gamma range activity and cortical blood flow. However, aminergic ascending neurons exert mainly diffuse inhibitory influences. These two kinds of influences together support reflective and rational psychological activities. During slow wave sleep, both kinds of ascending influences decrease and the mental content comprises low-intensity thought-like activities, similar to the waking mode of functioning, although dreams have been described. During rapid eye movement sleep, the principal dreaming stage, the cortex is activated but significantly disinhibited since all aminergic neurons are silent except the dopaminergic ones. We hypothesize that, in addition to this unusual state, the persistent release of dopamine associated with the specific silence of noradrenergic neurons could explain the characteristics of dream mental activity which are somewhat similar to psychotic symptoms. PMID- 11382894 TI - Sexually dimorphic effects of GHRH on sleep-endocrine activity in patients with depression and normal controls - part I: the sleep eeg. AB - In patients with depression, enhanced secretion of ACTH and cortisol, a reduction in slow wave sleep (SWS) and a blunted nocturnal growth hormone (GH) surge have been described and attributed, at least partly, to an elevation of corticotropin releasing hormone (CRH), hence a shift in the ratio between growth hormone releasing hormone (GHRH) and CRH. We investigated the effects of pulsatile administration of GHRH (4x50 microgram, at hourly intervals between 2200 and 0100 h) on the sleep EEG (2300-0700 h) in patients with depression (16 females, 19 males, age range 19-76 years) and matched controls (20 females, 20 males). In patients compared with controls, NREM sleep and in particular stage 2 sleep was greatly decreased at baseline. GHRH treatment enhanced NREM sleep, and in particular stage 2 sleep in men, regardless of diagnosis, while decreasing it in women (F=6.0 and 7.1, p<0.05). In controls, aging was associated with a decrease in NREM sleep, including both SWS and stage 2 sleep (r= -0.45 r= -0.39, p<0.05), while in patients only SWS declined with age (r= -0.49, p<0.05). The significant decrease in NREM sleep including stage 2 sleep in patients with depression and elderly control subjects is compatible with the suggested role of sleep continuity and stage 2 sleep in cognitive functioning. GHRH promoted NREM sleep, stage 2 sleep and sleep continuity and might prove beneficial for improvement of cognitive function, at least in men. These data support the hypothesis that female gender, aging and depression are associated with a shift in the GHRH/CRH ratio towards CRH. PMID- 11382895 TI - Sexually dimorphic effects of GHRH on sleep-endocrine activity in patients with depression and normal controls - part II: hormone secretion. AB - In depression and aging an increase in nocturnal cortisol secretion and a blunted nocturnal growth hormone (GH) surge have been described. In normal young men, growth hormone-releasing hormone (GHRH) promotes GH release and reduces plasma cortisol. Here, we examined whether GHRH could help to restore sleep-endocrine regulation in patients with depression and aging. GHRH (4x50 microgram, at 2200, 2300, 2400 and 0100 h) or saline (placebo) was injected intravenously to 42 patients with depression (19 females, 23 males) and matched controls (age range 19-76 years). Blood samples were withdrawn at 20 min intervals between 2200-0700 h and analysed using Manova (D.F. 1, 72). Patients compared to controls had significantly higher levels of ACTH and cortisol, particularly during the first half of the night (F=9 and F=11.8, each p<0.05). GHRH reduced ACTH during the first and cortisol secretion during the second half of the night in males, regardless of diagnosis, but enhanced it in females (F=5.1 and F=4.0, each p<0.05). ACTH and cortisol secretion were inversely related to NREM and stage 2 sleep in patients (r= -0.42, -0.42 and r= -0.36, -0.39, respectively, each p<0.05) but not in controls. Our data suggest that: 1) female gender, depression and aging add-on to enhance HPA activity, and 2) hyperactivity of the HPA system and the decrease in NREM and in particular stage 2 sleep in depression are interrelated. In men, GHRH can restore some of the sleep-endocrine alterations associated with depression and aging. PMID- 11382896 TI - Systemic administration of hypocretin-1 reduces cataplexy and normalizes sleep and waking durations in narcoleptic dogs. AB - Recent work has implicated the hypocretin (orexin) system in the genesis of narcolepsy. In the current study we demonstrate that systemically administered hypocretin-1 (Hcrt-1) produces an increase in activity level, longer waking periods, a decrease in REM sleep without change in nonREM sleep, reduced sleep fragmentation and a dose dependent reduction in cataplexy in canine narcoleptics. Repeated administration of single daily doses of Hcrt-1 led to consolidation of waking and sleep periods and to a complete loss of cataplexy for periods of three or more days after treatment in animals that were never asymptomatic under control conditions. Systemic administration of Hcrt-1 may be an effective treatment for narcolepsy. PMID- 11382897 TI - Dorsal raphe nucleus administration of 5-HT1A receptor agonist and antagonists: effect on rapid eye movement sleep in the rat. AB - The effect of flesinoxan, a selective 5-HT1A receptor agonist, WAY 100635, a selective 5-HT1A receptor antagonist, and (+/-)pindolol, a mixed beta adrenoceptor and 5-HT1A/B receptor antagonist, on spontaneous sleep was studied in adult rats implanted for chronic sleep recordings. Drugs were infused directly into the dorsal raphe nucleus (DRN). Direct application of flesinoxan (25.0 and/or 50.0 ng) into the DRN induced a significant increment of REM sleep (REMS) during the second and third 2 h period of recording. On the other hand, microinjection into the DRN of (+/-)pindolol (100.0 and/or 200.0 ng), and WAY 100635 (12.5, 25.0 and 50.0 ng) significantly reduced REMS during the first and/or second 2 h recording period. Our findings support previous studies indicating that microdialysis perfusion of the 5-HT1A receptor agonist 8-OHDPAT into the DRN increases REMS. In addition, they favor the proposal that microinjection of 5-HT1A receptor antagonists into the DRN would suppress 5-HT inhibition and reduce REMS. PMID- 11382898 TI - Distribution of hypocretin-containing neurons in the lateral hypothalamus and C fos-immunoreactive neurons in the VLPO. AB - The present study investigated the distribution of neurons implicated in the regulation of sleep in three species generally used in sleep research, i.e., mice, rats and cats. We focused on sleep active neurons in the ventral lateral preoptic (VLPO) area and the hypocretin/orexin-containing neurons in the lateral hypothalamus. The latter groups of neurons were found recently to play an important role in the regulation of REM sleep. The expression of the transcription factor, c-Fos, was used to identify the VLPO. In mice and rats, in response to sleep, a discrete cluster of c-Fos positive cells was found in the VLPO. In mice, this cluster was located more medially compared to the rat, and as in the rat, galanin immunostained neurons were found in the VLPO. In the cat, c Fos positive cells did not segregate to a specific location but were more diffusely represented in the preoptic area. In all three species, orexin/hypocretin-containing neurons were located only in the lateral hypothalamus with the distribution being more diffuse in the cat. The grouping of sleep-active cells in rodents makes it feasible to extract these cells for tissue culture and molecular analysis. Moreover, given that rodents have a distinct circadian distribution of sleep-wakefulness, the connectivity with the suprachiasmatic nucleus can also be determined. PMID- 11382899 TI - How significant are primary sleep disorders and sleepiness in the chronic fatigue syndrome? AB - In order to study both the prevalence of Primary Sleep Disorders (PSD) and sleepiness, and their association to the Chronic Fatigue Syndrome (CFS), 46 unselected outpatients (34 women, mean age 36.5) were examined clinically and underwent two nights of all-night polysomnography and multiple sleep latency tests (MSLT). Forty-six percent presented with a Sleep Apnea/Hypopnea Syndrome Index (AHI>=5), 5% with a Periodic Limb Movements syndrome. No subject received a diagnosis of Narcolepsy or Idiopathic Hypersomnia. Thirty percent showed the presence of objective sleepiness as measured by MSLT<10 minutes. Objective and subjective measures of sleepiness were not associated with CFS, nor with the double diagnosis of CFS and a PSD. The presence of PSD or sleepiness was not associated with any of the clinical scales that were used to measure anxiety, depression, somatisation, physical or mental fatigue, or functional status impairment. Fifty-four percent of CFS patients had no PSD, and 69% no sleepiness. These patients could not be distinguished clinically from patients having a PSD or from those with sleepiness. Therefore, it is unlikely that CFS is simply a somatic expression of any PSD observed in our sample or of sleepiness per se. PMID- 11382900 TI - Some basic features of the new sleep-aid tea (SAT) for the treatment of insomnia. AB - The "immediate-onset"-acting hypnotics (1-2 h after oral administration) (e.g., benzodiazepines) serve as the leading approach and great progress has been made in this century. They are exogenous from artificial synthesis and mainly fit for short-term insomnia. The "gradual-onset"-acting hypnotics (3-5 d after oral administration) are mild and gradually effective, but are very safe and without noticeable side or adverse effects. They are endogenous or endogenous-mimetic from natural isolation. It may serve as a more natural approach for the treatment of chronic or long-term insomnia, which is mainly gradually developed. This approach is especially useful in treating elderly people, whose numbers are rapidly increasing worldwide, especially in the next century. Sleep-Aid Tea (SAT) is a naturally processed Tea consisting of endogenous or endogenous-mimetic sleep inducing substances isolated from specific natural and edible fruits and plants well known for the treatment of insomnia, e.g., Compendium of Materia Medica (Li, 1596) associated with modern knowledge and isolation technology (Liu, 1990, 1993). SAT is especially fit for the treatment of insomnia in aged people because it is mild, "gradual-onset"-acting (3-5 d after oral administration), natural, safe and without side effects. PMID- 11382901 TI - Brainstem origin for a new very slow (1mHz) oscillation in the human non-REM sleep episode. AB - The time-courses of power in the different frequency bands (1-40 Hz) within the non-rapid-eye-movement (NREM) episode of the human sleep electroencephalogram have provided for many years a fascinating window into the sleep process. Here our analysis of the slow-wave band (1-4 Hz) reveals a hitherto unrecognized very slow oscillation of power with mean period ~15 minutes, an instability that appears to be an integral characteristic of the early NREM episode. The neuronal transition probability (NTP) model has already given a mechanism explaining how power in the spindle band peaks consistently before that of slow wave activity. Here we show that an extension of the model, with the hypothesis of a population of sleep neurons alternating between two steady probability states, can simulate the very slow oscillation. In doing so it gives not only the time course of power in the slow wave band, but also the simultaneous time-courses in the spindle and in the fast frequency bands. Animal data suggest that a brainstem neuronal population, toggled by an external switching source, generates these time-courses and dictates them to the thalamus and thence to the cortex. The discovery of the very slow oscillation and the success of the NTP model in interpreting the overall NREM structure may have important implications for both clinical and fundamental sleep research. PMID- 11382902 TI - Isolated sleep paralysis: a web survey. AB - Isolated Sleep Paralysis (SP) occurs at least once in a lifetime in 40-50% of normal subjects, while as a chronic complaint it is an uncommon and scarcely known disorder. A series of messages written by subjects who experienced at least one episode of SP, containing more or less detailed descriptions of this disorder, were collected from the Sleep Web site of the University of California in Los Angeles between January 1996 and July 1998. Two hundred and sixty-four messages fulfilling the International Classification of Sleep Disorders (ICSD) (Thorpy, 1990) minimal criteria for SP were analyzed. A wide spectrum of severity was evident, with a frequency of episodes ranging from one in a lifetime to almost every night, and a variety of emotional and hallucinatory experiences associated with SP episodes were reported. Clinical similarities between the recurrent form of isolated SP and channelopathies (in particular, periodic paralyses) are discussed. An activation of limbic system structures is suggested in order to explain some of the most common subjective experiences associated with SP. PMID- 11382903 TI - Slow wave and REM sleep mentation. AB - The aim of this experiment was to compare the characteristics of mental activity during REM and Slow Wave Sleep (SWS). Forty dream reports and their mnemonic associations were collected from twenty subjects. The reports were analyzed for structure (number of temporal units, number of report multi-units, and narrative continuity), awareness (reality testing, subjective time), and content (self, setting, laboratory references, number of non-self characters, implausibility, body feelings, number and intensity of emotions, vividness). Associations were classified as episodic and semantic memories. Results showed that REM reports were significantly longer than SWS reports. Minor content SWS-REM differences were also detected, which were more quantitative than qualitative. The collected observations might support the tentative hypothesis that dreaming is a continuous process, which is not unique to REM sleep. Different levels of engagement of the cognitive system are responsible for the SWS-REM differences that were detected. PMID- 11382904 TI - A polymorphism in the human timeless gene is not associated with diurnal preferences in normal adults. AB - The effect of a single nucleotide polymorphism, a glutamine to arginine amino acid substitution in the human Timeless gene (Q831R, A2634G), on diurnal preferences was studied in a random sample of normal volunteers enrolled in a population-based epidemiology study of the natural history of sleep disorders. We genotyped 528 subjects for this single nucleotide polymorphism and determined morningness-eveningness tendencies using the Horne-Ostberg questionnaire. Our results indicate that Q831R Timeless has no influence on morningness- eveningness tendencies in humans. PMID- 11382905 TI - Cerebral circulation in REM sleep: is oxygen a main regulating factor? AB - In the transition from NREM to REM sleep, as in other instances of brain activation, a marked increase in cerebral blood flow and glucose uptake is observed, together with a lesser increase in O2 uptake. Brain activation also entails an increase in capillary PO2 and lactate production. The hypothesis of saturation of the oxidative machinery was advanced to explain anaerobic glycolysis and lactate production in the presence of high PO2, but data are available that cannot be explained by this hypothesis: hypoxic spots exist in the brain, augmenting in arterial hypoxia and disappearing in arterial hyperoxia, while tissue [H+] lowers as arterial PO2 increases beyond 100 mmHg. Additional hypotheses are thus required. We suggest that O2 diffusion limitation exists in the brain: microregions lying at mid-distance between capillaries may become hypoxic and partly resort to anaerobic glycolysis. These microregions are thought to enlarge with increasing metabolic rate or arterial hypoxia and give rise to vasodilatatory signals regulating local blood flow. REM sleep time is strongly reduced by hypoxic and increased by hyperoxic atmosphere, in accordance with the existence of an O2 diffusion limitation. Any pathological decrease in arterial PO2 and/or O2 delivery creates a specific risk in REM sleep. PMID- 11382906 TI - Sleep/wake cycle and physical activity in healthy elderly people. AB - Investigations concerning the temporal organization of the sleep/wake cycle (SWC) in healthy active elderly, specially focusing possible relationships between regular daily routines, sleep fragmentation and sleep quality in aging are still scarce. We studied the SWC of 23 healthy active elderly (65-76 years old) of both sexes (15 female, 8 male), engaged in an exercise program for old people, considering that physical exercise sessions could represent a strategy for keeping structured daily routines. We report the relationships between SWC characteristics and performance of physical exercises viewed as time cues. We collected SWC data by means of sleep logs and data about light exposition by means of actigraphy during 23 consecutive days. Individuals who chose earlier times for exercising showed longer exposition to natural bright light and reported better sleep quality. Indications that physical exercises could be beneficial upon sleep were scarce (better subjective sleep quality). Strength of semicircadian component of the SWC was greater for subjects who exercised longer, suggesting they have advantageously used naps as part of their active and healthy lifestyle, and indicating the need for studying more closely the temporal organization of naps. PMID- 11382907 TI - Effect of age on sensory gating of the sleep state-dependent P1/P50 midlatency auditory evoked potential. AB - The P1/P50 midlatency auditory evoked potential is a sleep state-dependent waveform present during waking and rapid eye movement (REM) sleep and absent during slow-wave sleep. The P50 potential was studied in normal male and female subjects of various ages including post-pubertal adolescents (12-19 yrs), young adults (24-39 yrs), middle-aged adults (40-55 yrs) and older adults (55-78 yrs). There were no statistically significant differences in the mean peak amplitude or mean peak latency of the P50 potential between males and females or between age groups. Using a paired stimulus paradigm, the degree of sensory gating of the P50 potential was tested at three different interstimulus intervals (ISIs), 250, 500 and 1000 msec. There were no statistically significant differences in the sensory gating of the P50 potential between males and females. However, there was a significant decrease in sensory gating of the P50 potential in the adolescent group compared to each of the other age groups at the 250 msec ISI, but not at the 500 or 1000 msec ISI. These results suggest the presence of decreased sensory gating in normal adolescents compared to normal, older age groups. PMID- 11382908 TI - Timing patterns of cluster headaches and association with symptoms of obstructive sleep apnea. AB - Cluster headaches (CH) frequently recur at the same point in the circadian cycle, often during sleep. They may, in some cases, represent a susceptible individual's response to hypoxemia or other physiological changes induced by obstructive sleep apnea (OSA). If and when this mechanism exists, timing of CH close to the onset of sleep-and therefore OSA-might be expected. We questioned 36 subjects with CH about the times at which their CH usually occurred and about several symptoms known to be predictive of OSA, including habitual snoring, loud snoring, observed apneas and excessive daytime sleepiness. We then used logistic regression to determine whether occurrence of CH in each of six time periods was associated with OSA symptoms. The 23 subjects (64%) who reported CH in the first half of a typical night's sleep also tended to report headaches during the midday/afternoon period. Symptoms of OSA, and in particular habitual snoring, were predictive of both first-half-of-the-night and midday/afternoon CH (p<.05). Thirty-one subjects (86%) reported that their CH were sleep-related, usually occurring during any part of the night or on awakening, but symptoms of OSA were not predictive of this timing pattern. In short, several OSA symptoms showed an association with CH occurrence in the first half of the night but not with sleep-related CH in general. These findings suggest that in some patients, physiological consequences of OSA may trigger CH during the first few hours of sleep and thereby influence the timing of subsequent daytime headaches. PMID- 11382909 TI - EEG delta activity during undisturbed sleep in the squirrel monkey. AB - The squirrel monkey (Saimiri sciureus) exhibits a robust daily rhythm of sleep wakefulness that is under circadian control, but the nature of homeostatic sleep regulation in this diurnal primate is poorly understood. Since delta frequency (0.5-2.0 Hz) activity in the electroencephalogram (EEG) during non-Rapid Eye Movement (NREM) sleep is thought to reflect homeostatic factors contributing to sleep tendency, we measured EEG delta power density and slow wave incidence and amplitude during NREM sleep during spontaneous sleep, occurring when monkeys were housed undisturbed in a 24-hour light-dark (LD) cycle and in constant light (LL). In LD and LL conditions, monkeys exhibited circadian rhythms in delta power density, wave incidence and wave amplitude that peaked in the middle of the subjective night, several hours after consolidated sleep onset. These results differ from predictions of a purely homeostatic model of sleep that would include maximal levels of delta activity at sleep onset. PMID- 11382910 TI - Exercise training effect on obstructive sleep apnea syndrome. AB - The role, if any, of exercise training in the management of individuals with obstructive sleep apnea syndrome (OSAS) is unclear. Anecdotally, patients have reported improvement in symptoms with regular participation in an exercise regime. In this study, we evaluated the effects of an exercise training program and weight loss on physical and subjective measures associated with OSAS. Nine subjects with mild to moderate OSAS completed a six month supervised exercise program. Pre and post-training measures on polysomnographic testing, physical training, anthropometric measures, quality of life (QOL), daytime somnolence and mood states were assessed. A significant decrease in the AHI (p=0.002) was noted along with improvements (p<0.05) in total sleep time, sleep efficiency, number of awakenings/hour, arousals/hour, apnea index and mean exercise training workloads. Significant decreases (p<.001) in weight (-6.2 kg) and body mass index (-1.6) were observed. Evaluation of QOL measures by the Health Status Questionnaire, Profile of Mood States and Epworth Sleepiness Scale showed significant changes in health status, affective state, and a decrease in daytime somnolence. Regular exercise training had a positive impact on the AHI, aerobic capacity, body mass index and QOL. However, exercise training alone was not an adequate intervention strategy for most individuals with OSAS but may serve well as an adjunct treatment strategy in the conservative management of individuals with mild to moderate OSAS. PMID- 11382912 TI - Effect of ambient temperature on sleep-wakefulness in normal and medial preoptic area lesioned rats. AB - The changes in sleep-wakefulness were studied in rats during their exposure to different ambient temperatures of 18 degrees C, 24 degrees C and 30 degrees C, before and after the destruction of the medial preoptic area neurons by N-Methyl D-aspartic acid. In normal rats, there was an increase in paradoxical sleep and slow wave sleep and a decrease in wakefulness at higher ambient temperatures. The increase in sleep was primarily due to an increase in the duration of sleep episodes. Destruction of the medial preoptic area neurons produced a decrease in sleep at all three different ambient temperatures. But, there was a linear increase in sleep with higher temperatures in the lesioned rats that was qualitatively different from that in the normal animals, as the increase in sleep was associated with an increase in the number of short duration slow wave sleep episodes. The findings indicate that the medial preoptic area is essential for sleep maintenance and improving the quality of sleep with higher ambient temperatures. It is possible that the medial preoptic area serves as a fine tuning mechanism to regulate sleep for energy homeostasis, including thermoregulation. PMID- 11382911 TI - Restorative effects of a short afternoon nap (<30 min) in the elderly on subjective mood, performance and eeg activity. AB - This study investigated the effects of a short afternoon nap (<30 min) in the elderly on subjective mood, performance and electroencephalograms (EEG). Ten healthy elderly persons who habitually napped in the afternoon three or more times a week participated in the present study. They participated in two experimental conditions with an interval of more than five days. In the nap condition, the participants went to bed at 13:00 hours and slept for 30 min, whereas they just had a rest while watching television in the no-nap condition. In both conditions, subjective sleepiness, fatigue, performance and EEG activities with eyes opened and closed were repeatedly measured before and after the nap or rest. The nap significantly reduced subjective sleepiness and fatigue in the afternoon. It also improved performance levels and EEG theta, alpha 1 or alpha 2 band activities with eyes opened and closed. These findings suggest that a short afternoon nap is useful for the elderly in maintaining their daytime psychological, behavioral and physiological arousal at an adequate level. PMID- 11382913 TI - State-dependency of neuronal slow dynamics during sleep observed in cat lateral geniculate nucleus. AB - From the accumulated results, we hypothesize that neurons in the central processor systems of the brain generally exhibit a common state-dependency in slow dynamics of their spontaneous activities during sleep. In this paper, activities of relay cells in the cat's lateral geniculate nucleus (LGN) were studied to see if our hypothesis can be applied in this thalamic region. Data segments in polygraphically steady states were strictly extracted in order to sample the activities whose stationarity was guaranteed in a statistical sense. During slow wave sleep (SWS), the discharge pattern was characterized by short bursts. In contrast, the rather tonic discharge pattern was observed to prevail during rapid eye movement (REM) sleep. Spectral analyses showed white noise-like spectra in the low frequency range of 0.04-1.0 Hz during SWS, and 1/f noise-like spectra in the same frequency range during REM sleep. This state-dependency of the slow dynamics was consistently characterized by the other statistical parameters concerning the second-order moment as well. In contrast, the fast dynamics over 1.0 Hz tended to exhibit neuron-specific changes associated with the sleep state in terms of the Markovian dependency analysis. Consequently, our working hypothesis was not rejected for the LGN relay cells. The result here extends the possibility that the state-dependency of the slow dynamics we found is a general rule concerning single neuronal dynamics in widespread areas of the brain during sleep. The state-dependency of the slow dynamics of the LGN relay cells could be understood according to the proposed mechanism that a state associated alteration in the global biasing input to a neural network during sleep induces the phenomenon with which we are concerned. The slow dynamics of neuronal activities might provide a novel framework defining SWS and REM sleep states instead of the polygraphic characteristics. PMID- 11382914 TI - Topographical distribution of spindles: variations between and within nrem sleep cycles. AB - Spindle density, visually scored in the 12-15 Hz range over antero-posterior midline derivations, was assessed during a baseline night in ten normal subjects. Sleep spindles were found to be highly variable between subjects and more abundant during Stage 2. Topographical distribution of spindle density showed a centroparietal prevalence, stable between NREM sleep stages. Intra-night variations of spindle density exhibited a linear increase across consecutive NREM episodes, suggesting an inverse relation with the time course of slow wave sleep. Except for occipital leads reaching a maximum during the third NREM cycle and then decreasing, changes in spindle density across sleep cycles were similar over different derivations. Intra-cycle variations fit a fourth-order polynomial curve with a minimum in the middle part of each sleep episode (when most slow wave sleep is expressed); this intra-cycle trend also seems stable between derivations and consecutive sleep cycles. These results confirm and extend, to the level of macroscopic EEG, the reciprocal relationship between sigma and delta waves previously shown by spectral analysis of EEG frequencies and, at a neuronal level in the thalamocortical network, by changes of membrane potentials that oscillate in the frequency range of spindles or delta at different levels of hyperpolarization. PMID- 11382915 TI - Altered sleep and behavioral patterns of arthritic rats. AB - The present study sought to evaluate concomitant alterations of behavioral and sleep patterns of arthritic rats. Rats were implanted with electrodes for polysomnographic recordings and submitted to the model of arthritis by a subcutaneous (s.c.) administration of Freund adjuvant in the posterior right paw and saline in the posterior left paw. The SHAM group was injected with saline in both paws, whereas the control group (CTL) was not submitted to any manipulation. Behavioral tests were carried out twice before induction of arthritis, on the second day of arthritis, and once a week afterwards until the eighth week. Body weight, colonic temperature, and measurements of the injured paw were carried out on the same days. Arthritic rats presented a reduction of total sleep time, increased latency to synchronized sleep, augmented number of episodes of synchronized sleep, reduction of sleep efficiency, more stage shifts, and increased total alert time. Moreover, these animals presented a lower pain threshold than control and SHAM animals. This reduction was observed on the second day of arthritis and remained so reduced until the end of the study. The data appear to indicate a relationship between altered sleep pattern and increased pain sensitivity in arthritic rats. PMID- 11382916 TI - The relationship between esophageal pressure and apnea hypopnea index in obstructive sleep apnea-hypopnea syndrome. AB - Severity of negative esophageal pressure (Pes) and apnea hypopnea index (AHI) were investigated in 34 patients with obstructive sleep apnea-hypopnea syndrome (OSAHS). The OSAHS patients were diagnostically classified as having obstructive sleep apnea syndrome (OSAS) or upper airway resistance syndrome (UARS). Diagnosis of OSAS was based on an AHI of more than 5, and that of UARS on an AHI of less than 5, EEG arousals which were associated with apnea, hypopnea and/or respiratory effort occurring more than 10 times per hour, and daytime sleepiness. Negative Pes was represented by the greatest peak (NPes Max) and the number of increased (more than 13.5 cmH2O) episodes per hour (NPesI13.5). There was no significant correlation between the AHI and Pes indices, but NPes Max and NPesI13.5 showed significant correlation (p<0.01). NPes Max and NPesI13.5 showed no significant differences among the severe OSAS (AHI>50; 8 cases), moderate OSAS (50>AHI>15; 10 cases), mild OSAS (15>AHI>5; 9 cases) and UARS (7 cases) groups. We conclude that AHI does not reflect the severity of the increase in negative Pes, which is an important aspect of the pathophysiology of OSAHS. Assessment of OSAHS based on AHI alone may therefore underestimate the risk of increased negative Pes in cases with reduced AHI. PMID- 11382917 TI - Endoglin: An accessory component of the TGF-beta-binding receptor-complex with diagnostic, prognostic, and bioimmunotherapeutic potential in human malignancies. AB - Endoglin (CD105) is a cell membrane glycoprotein over-expressed on highly proliferating endothelial cells in culture, and on endothelial cells of angiogenetic blood vessels within benign and malignant tissues. CD105 binds several factors of the Transforming Growth Factor (TGF)-beta superfamily, and its over-expression modulates cellular responses to TGF-beta1. The complex of experimental findings accumulated in the last few years strongly indicate that CD105 is a powerful marker of angiogenesis, and that it might play a critical role in the pathogenesis of vascular diseases and in tumor progression. In this paper, we will review the structural, biological and functional features of CD105, as well as its distribution within normal and neoplastic tissues, emphasizing its foreseeable role as a molecular target for new diagnostic and bioimmunotherapeutic approaches in human malignancies. PMID- 11382918 TI - Early molecular changes in irradiated aortic endothelium. AB - Irradiated aortic endothelial cells (EC) exhibit distinct morphological, functional, and physiological responses to ionizing radiation (IR). However, the molecular basis for these responses has not been fully characterized. Cultured bovine and rat aortic endothelial cells were exposed to single fraction doses (0 30 Gy) of gamma radiation. IR caused dose-dependent DNA strand breaks which were repaired to near baseline levels within 30 min. A dose-dependent inhibition of cell growth was noted for IR greater than 1 Gy. At doses greater than 2.5 Gy, morphologic changes consistent with apoptosis and loss of cell viability were present beginning 12-16 h after radiation, with subsequent detachment of EC from the cell monolayer. By Western blot analysis, expression of p53, gadd45, p21, and bax protein increased in a time-and dose-dependent manner; p53 expression was maximal at 3 h after IR, and gadd45, bax and p21 levels peaked at 6 h. By Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), levels of p53 mRNA were not significantly increased after IR, whereas gadd45 exhibited time- and dose dependent increase in mRNA synthesis after IR. Activation of intracellular caspases, manifest by proteolytic poly (ADP-ribose) polymerase (PARP) and lamin B cleavage, was maximal at 15 h after IR, concident with other indices of EC apoptosis, including oligonucleosomal DNA degradation, TUNEL immunostaining, and morphologic changes. The tripeptide protease inhibitor z-Val-Ala-Asp (zVAD) prevented PARP and lamin cleavage, DNA fragmentation, morphological changes, and cell detachment in irradiated EC. The combined data suggested that gamma radiation induces a dose- and time-dependent sequence of early events in cultured EC with modulate growth arrest, apoptosis, and possibly premature senescence in surviving cells. PMID- 11382919 TI - Phosphorylation of focal adhesion kinase (pp125(FAK)) is increased in human keratinocytes induced to migrate by extracellular matrices. AB - During the healing process of skin wounds, human keratinocytes migrate across a provisional matrix of the wound bed. The mechanisms by which keratinocytes migrate on connective tissue are not known. In this study, we examined the role of focal adhesion kinase (FAK), an 125 kDa protein that co-localizes with focal adhesions in cells plated on extracellular matrix. We induced human keratinocytes into various states of migration by plating them on extracellular matrices that minimally, moderately, or strongly induce cellular migration, and then examined the expression of FAK at the protein level and its degree of tyrosine phosphorylation using Western immunoblotting and immunoprecipitation. In highly migratory human keratinocytes, we found that three proteins were predominantly tyrosine phosphorylated, one of them being FAK. Tyrosine phosphorylation of FAK tightly correlated with the level of cellular motility but not cell attachment to the matrix. Time course experiments demonstrated that in highly motile keratinocytes, tyrosine phosphorylation of FAK peaked at 12 h, the time when maximal migration on the matrix ensues. In contrast to FAK, the beta1 integrin subunit of human keratinocytes that configures with the alpha2, alpha3, and alpha5 integrin subunits to form integrin receptors for matrix, did not display tyrosine phosphorylation linked to motility. Using anti-sense oligonucleotides to FAK, we demonstrate that FAK is required for human keratinocyte migration, but not for focal adhesion formation. PMID- 11382920 TI - Hypoxia-induced upregulation of eNOS gene expression is redox-sensitive: a comparison between hypoxia and inhibitors of cell metabolism. AB - Several papers report a hypoxia-induced upregulation of the endothelial nitric oxide synthase (eNOS) mRNA expression. Since there is no known hypoxia-sensitive element binding site in the eNOS promoter, we reasoned that the effect of hypoxia could be simulated by a metabolically elicited alteration of the redox state. Therefore, cultured porcine aortic endothelial cells (PAEC) were exposed to hypoxia (1-10% O(2)) or inhibitors of cellular energy metabolism including rotenone, 2, 4 dinitrophenol (DNP) and 2-deoxyglucose for 6 to 24 h. Additionally, cells were treated with lactate and nicotinic acid to alter the cellular NAD(P)H/NAD(P) ratio without changes of energy supply. The cellular NAD(P)H/NAD(P) ratio was used as an index of the cellular redox state and determined using the MTT-assay. Hypoxia increased eNOS mRNA transcription and MTT reduction in a manner inversely proportional to pO(2). Exposure to rotenone, DNP, and lactate increased the NAD(P)H/NAD(P) ratio, MTT-reduction, and eNOS mRNA also in parallel. In contrast, 2-deoxyglucose and nicotinic acid attenuated both MTT reduction and eNOS mRNA expression. In order to study a potential role of the redox regulated transcription factor complex AP-1 in hypoxia-induced eNOS mRNA transcription, c-jun expression was determined and decoy experiments were performed. c-jun expression paralleled changes of eNOS mRNA expression and MTT reduction. Furthermore, in the presence of oligodeoxynucleotides corresponding to the AP-1 binding sites of the eNOS promoter, the hypoxia and chemically induced eNOS mRNA expression was completely abolished. We propose that hypoxia, by altering cellular metabolism, leads to an increase in the cellular NAD(P)H/NAD(P) ratio which favors enhanced eNOS expression by redox-sensitive AP-1 mediated transcriptional control. PMID- 11382921 TI - COP-1, a member of the CCN family, is a heparin-induced growth arrest specific gene in vascular smooth muscle cells. AB - Vascular smooth muscle cell (VSMC) hyperplasia is responsible for the failure of 15-30% of vascular surgical procedures such as coronary artery bypass grafts and angioplasties. We and others have shown that heparin suppresses VSMC proliferation in vivo and in cell culture. We hypothesize that heparin inhibits VSMC proliferation by binding to cell surface receptors, resulting in selective modulation of mitogenic signal transduction pathways and altered transcription of a specific subset of growth regulatory genes. To test this idea, we used subtractive hybridization to identify differentially expressed mRNAs in heparin treated and untreated VSMC. We identified a heparin induced mRNA identical to Cop 1, a member of the CCN family of proteins which are secreted, cysteine-rich modular proteins involved in growth regulation and migration. Cop-1 from smooth muscle cells appears to have a different expression pattern and possibly different functions than Cop-1 from other cells. Cop-1 mRNA is expressed at high levels in quiescent VSMC and at low levels in proliferating VSMC, an expression pattern highly characteristic of growth arrest specific genes. Cop-1 mRNA is expressed at high levels in heparin treated VSMC and COP-1 protein is secreted into culture medium. In tissues, Cop-1 expression is observed in the uninjured rat aorta suggesting a possible role for Cop-1 in vivo. We found PDGF, but not EGF, inhibits the expression of Cop-1 in VSMC. Neither TGF-beta nor interferon beta, two inhibitors of VSMC proliferation, were able to induce Cop-1 expression. In addition, heparin does not induce Cop-1 mRNA in endothelial cells and VSMC resistant to the antiproliferative effect of heparin. Conditioned medium from cells over-expressing COP-1 protein inhibits VSMC proliferation in culture. Together, our data indicate that COP-1 may play a role in the antiproliferative mechanism of action of heparin. PMID- 11382922 TI - Reduction of c-myc expression by an antisense approach under Cre/loxP switching induces apoptosis in human liver cancer cells. AB - c-Myc has been documented to be both a positive and a negative signal for the induction of apoptosis. It is well known that overexpression of the c-myc gene induces apoptosis of normal cells, but the result of a reduction in its expression is not fully understood. We examined whether a reduction in c-myc expression would induce apoptosis in human liver cancer cells. Specifically, antisense and sense oligodeoxynucleotides (oligos) against the human c-myc mRNA were synthesized, mixed with a liposome reagent at various ratios, and were applied to the liver cancer-derived cell lines, HCC-T, HepG2, and PLC/PRF/5. To exclude effects resulting from using oligos, plasmid vectors expressing the full length c-myc cDNA in both sense and antisense orientations under the control of the Cre/loxP system were generated. Monoclonal cell lines including these plasmid vectors were produced and Cre was supplied by adenovirus infection. Apoptosis was determined morphologically and c-Myc and Bcl-2 expression was examined by Western blotting. The antisense myc significantly inhibited the proliferation of the cells within two days, while neither the liposome reagent alone nor sense myc did so. Most of the cells were rounded up by the antisense-treatment and nuclear fragmentation and DNA ladder formation were detected after two days in antisense c-myc-treated cells. Antisense c-myc largely reduced c-Myc and partially Bcl-2 expression; overexpression of Bcl-2 partially rescued from apoptosis in HCC-T and HepG2 cells. These results suggest that the massive reduction in c-myc mRNA induces apoptosis in liver cancer cell lines and consequent decrease in Bcl-2 enhances the cell death. c-Myc reduction under the Cre/loxP switching system may be a useful tool for the clarification of c-myc-related cellular mechanisms in differentiation and proliferation. PMID- 11382923 TI - Perlecan inhibits smooth muscle cell adhesion to fibronectin: role of heparan sulfate. AB - Smooth muscle cell migration, proliferation, and deposition of extracellular matrix are key events in atherogenesis and restenosis development. To explore the mechanisms that regulate smooth muscle cell function, we have investigated whether perlecan, a basement membrane heparan sulfate proteoglycan, modulates interaction between smooth muscle cells and other matrix components. A combined substrate of fibronectin and perlecan showed a reduced adhesion of rat aortic smooth muscle cells by 70-90% in comparison to fibronectin alone. In contrast, perlecan did not interfere with cell adhesion to laminin. Heparinase treated perlecan lost 60% of its anti-adhesive effect. Furthermore, heparan sulfate as well as heparin reduced smooth muscle cell adhesion when combined with fibronectin whereas neither hyaluronan nor chondroitin sulfate had any anti adhesive effects. Addition of heparin as a second coating to a preformed fibronectin matrix did not affect cell adhesion. Cell adhesion to the 105- and 120 kDa cell-binding fragments of fibronectin, lacking the main heparin-binding domains, was also inhibited by heparin. In addition, co-coating of fibronectin and (3)H-heparin showed that heparin was not even incorporated in the substrate. Morphologically, smooth muscle cells adhering to a substrate prepared by co coating of fibronectin and perlecan or heparin were small, rounded, lacked focal contacts, and showed poorly developed stress fibers of actin. The results show that the heparan sulfate chains of perlecan lead to altered interactions between smooth muscle cells and fibronectin, possibly due to conformational changes in the fibronectin molecule. Such interactions may influence smooth muscle cell function in atherogenesis and vascular repair processes. PMID- 11382924 TI - Functional role of matrix metalloproteinases (MMPs) in mammary epithelial cell development. AB - The extracellular matrix (ECM) is an important regulator of mammary epithelial cell (MEC) function and is remodeled by matrix metalloproteinases (MMPs). To investigate the significance and regulation of MMP activity in normal MEC, we utilized a primary culture model in which rat MEC were grown three dimensionally within a reconstituted basement membrane (RBM) in defined serum-free medium. Zymograms of culture medium demonstrated that five major gelatinases of 97, 80, 74, 69, and 65 kDa were secreted by MEC and were distinct from gelatinases of RBM origin. Based on molecular weight, p-aminophenylmercuric acid activation, immunoblotting with MMP-specific antibodies, inhibition by EDTA, a peptide containing the prodomain sequence of MMP (TMRKPRCGNPDVAN) and two synthetic MMP inhibitors (BB-94 and CGS 27023A), these were classified as inactive and active forms of MMP-9 and MMP-2. The maximal MMP activities occurred when MEC were in a rapid proliferation and branching phase and declined after they underwent functional differentiation. Known regulators of MEC growth and differentiation were evaluated for their ability to modulate gelatinase activity in primary culture. Secretion of one or both MMPs was inhibited by EGF, TGFalpha, prolactin, and hydrocortisone and stimulated by progesterone. Furthermore, the functional significance of MMPs was demonstrated since three MMP inhibitors blocked branching morphogenesis elicited by the absence of hydrocortisone. Additionally, two synthetic MMP inhibitors not only inhibited epithelial cell growth but also inhibited normal alveolar development of the MEC. Finally, these drugs were found to enhance MMP secretion from MEC, although the activity of the secreted MMPs was inhibited as long as the drug was present. PMID- 11382925 TI - Carbonic anhydrase II is an AP-1 target gene in osteoclasts. AB - c-Fos, a member of the AP-1 family of transcription factors, is necessary for osteoclast differentiation but to date, none of the osteoclast-phenotypic markers have been identified as AP-1 target genes. Here, we demonstrate that carbonic anhydrase II (CA II), an enzyme necessary for osteoclast activity, is transcriptionally upregulated by c-Fos/AP-1. A functional AP-1 binding site is present in the CA II promoter and is necessary for this regulation. Furthermore, we show that AP-1 binding activity, mainly composed of Fra-2 and JunD, is induced by treatment of bone marrow cultures with the osteoclastogenic hormone 1,25 dihydroxyvitamin D(3). Fra-2 and JunD are found in mature osteoclasts as well. Thus, our data demonstrate that cFos/AP-1 can directly regulate the expression of this osteoclast marker and that AP-1 activity is upregulated in osteoclast progenitors in response to osteoclastogenic signals. PMID- 11382926 TI - Evidence that the death receptor DR4 is a DNA damage-inducible, p53-regulated gene. AB - DR4 (TRAIL-R1), a member of the tumor necrosis factor receptor superfamily, is a cell surface receptor that triggers the apoptotic machinery upon binding to its ligand tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). Although three other TRAIL receptors DR5, DcR1, and DcR2 are induced by DNA damage and are regulated by the wild-type p53 tumor suppressor, it was not known whether these factors also affect DR4 expression. In this study, we found that DR4 expression is also enhanced by DNA damage whether induced by ionizing radiation or by chemotherapeutic agents. The induction was observed predominantly in cells containing wild-type p53 and was similar to the regulation patterns of DR5 and Fas, two other members of the family which are known to be regulated by p53. Transfection of HPV 16 E6 gene into cells with wild-type p53, which decreased the level of p53 protein, resulted in suppression of DR4 induction by DNA-damaging agents. Conversely, introduction of exogenous wild-type p53 through adenovirus infection has led to upregulation of endogenous DR4 in cells with mutant p53. Moreover, the transcription inhibitor actinomycin D abolished DNA-damaging agent induced DR4 expression. Thus, DR4 appears to be a DNA damage-inducible, p53 regulated gene. PMID- 11382928 TI - TNFalpha induces NFkappaB/p50 in association with the growth and morphogenesis of normal and transformed rat mammary epithelial cells. AB - In contrast to the cytotoxic or cytostatic effect of TNFalpha on many breast cancer cell lines, TNFalpha stimulates growth and morphogenesis of normal rat mammary epithelial cells (MEC). The present studies were carried out to determine whether there are intrinsic differences between normal and malignant MEC which may explain the differing responsiveness to TNFalpha. Freshly isolated rat MEC organoids from normal mammary gland or 1-methyl-1-nitrosourea-induced mammary tumors were treated with TNFalpha for 21 days. Unexpectedly, TNFalpha stimulated growth and morphogenesis of both normal and transformed MEC in primary culture, although in transformed cells its effects were delayed and the majority of the colonies were histologically abnormal, with multiple cell layers and no lumen. Since NFkappaB is a key mediator of TNFalpha action and has been implicated in carcinogenesis, the expression of the p50, p52, p65, and c-rel NFkappaB proteins in normal and transformed MEC was determined. Expression of p52 was significantly reduced in tumor cells, and p50 was absent, although its putative precursor, p105 was abundant. There were no changes in the levels of p65 or c-rel. TNFalpha induced a pronounced and sustained increase of a p50 homodimeric NFkappaB/DNA complex in both normal and transformed MEC. However, in transformed MEC, NFkappaB binding was initially undetectable but then increased in response to TNFalpha. Thus, NFkappaB expression and DNA binding activity are altered during mammary carcinogenesis. In addition, the significant increase in NFkappaB/p50 DNA-binding was temporally coincident with TNFalpha-induced growth and morphogenesis, suggesting that it may play a significant role in both normal development and carcinogenesis. PMID- 11382927 TI - Calyculin-A induces focal adhesion assembly and tyrosine phosphorylation of p125(Fak), p130(Cas), and paxillin in Swiss 3T3 cells. AB - Treatment of intact Swiss 3T3 cells with calyculin-A, an inhibitor of myosin light chain (MLC) phosphatase, induces tyrosine phosphorylation of p125(Fak) in a sharply concentration- and time-dependent manner. Maximal stimulation was 4.2 +/- 2.1-fold (n = 14). The stimulatory effect of calyculin-A was observed at low nanomolar concentrations (<10 nM); at higher concentrations (>10 nM) tyrosine phosphorylation of p125(Fak) was strikingly decreased. Calyculin-A induced tyrosine phosphorylation of p125(Fak) through a protein kinase C- and Ca(2+) independent pathway. Exposure to either cytochalasin-D or latrunculin-A, which disrupt actin organization by different mechanisms, abolished tyrosine phosphorylation of p125(Fak) in response to calyculin-A. Treatment with high concentrations of platelet-derived growth factor (20 ng/ml) which also disrupt actin stress fibers, completely inhibited tyrosine phosphorylation of p125(Fak) in response to calyculin-A. This agent also induced tyrosine phosphorylation of the focal adhesion-associated proteins p130(Cas) and paxillin. These tyrosine phosphorylation events were associated with a striking increase in the assembly of focal adhesions. The Rho kinase (ROK) inhibitor HA1077 that blocked focal adhesion formation by bombesin, had no effect on the focal adhesion assembly induced by calyculin-A. Thus, calyculin-A induces transient focal adhesion assembly and tyrosine phosphorylation of p125(Fak), p130(Cas), and paxillin, acting downstream of ROK. PMID- 11382930 TI - Association for Molecular Pathology: Sixth annual meeting in Denver, November 9 12, 2000. PMID- 11382929 TI - Regulation of angiotensin II-stimulated osteopontin expression in cardiac microvascular endothelial cells: role of p42/44 mitogen-activated protein kinase and reactive oxygen species. AB - Using spontaneously hypertensive and aortic banded rats, we have shown that expression of myocardial osteopontin, an extracellular matrix protein, coincides with the development of heart failure and is inhibited by captopril, suggesting a role for angiotensin II (ANG II). This study tested whether ANG II induces osteopontin expression in adult rat ventricular myocytes and cardiac microvascular endothelial cells (CMEC), and if so, whether induction is mediated via activation of mitogen-activated protein kinases (p42/44 MAPK) and involves reactive oxygen species (ROS). ANG II (1 microM, 16 h) increased osteopontin expression (fold increase 3.3+/-0.34, n = 12, P < 0.01) in CMEC as measured by northern analysis, but not in ARVM. ANG II stimulated osteopontin expression in CMEC in a time- (within 4 h) and concentration-dependent manner, which was prevented by the AT1 receptor antagonist, losartan. ANG II elicited robust phosphorylation of p42/44 MAPK as measured using phospho-specific antibodies, and increased superoxide production as measured by cytochrome c reduction and lucigenin chemiluminescence assays. These effects were blocked by diphenylene iodonium (DPI), an inhibitor of the flavoprotein component of NAD(P)H oxidase. PD98059, an inhibitor of p42/44 MAPK pathway, and DPI each inhibited ANG II stimulated osteopontin expression. Northern blot analysis showed basal expression of p22phox, a critical component of NADH/NADPH oxidase system, which was increased 40-60% by exposure to ANG II. These results suggest that p42/44 MAPK is a critical component of the ROS-sensitive signaling pathways activated by ANG II in CMEC and plays a key role in the regulation of osteopontin gene expression. Published 2001 Wiley-Liss, Inc. PMID- 11382931 TI - Structural characterisation of Baltic amber and its solvent extracts by several mass spectrometric methods. AB - A sample of Baltic amber ( approximately 40 million yrs old) has been extracted using pentane, toluene and 1-methyl-2-pyrrolidinone (NMP). The relationship between solubility characteristics of the extracts in relation to molecular mass and chemical makeup has been investigated. The extracts were first characterised by (13)C-NMR spectrometry, size exclusion chromatography (SEC) and UV fluorescence spectroscopy. The fractions differed less in terms of chemical structural features than they did in terms of molecular mass. This contrasts markedly with data on fractions of coal-derived liquids, but parallels results from petroleum-derived vacuum residues. In SEC, the toluene soluble/pentane insoluble fraction gave a peak for high mass material at about 67 000 u. Material excluded from the column porosity in this fraction and in NMP solubles eluted between 8 and 11 min, corresponding to polystyrene masses between 200 000 and several million u. A column with a larger pore size distribution was calibrated using polystyrene and polymethylmethacrylate standards with detection by a light scattering evaporative analyser. The largest polystyrene standard (15.4 million u) eluted at 13.4 min, similar to that of the earliest eluting amber-derived material in the NMP solubles fraction. Results from probe-MS and pyrolysis-GC/MS have been used to confirm the similarity of chemical structures of the three solubility fractions. Broadly, low mass ions appear to correspond to the various monomeric units of structures present in the amber, the higher mass ions to dimer units and the molecular ions to the different combinations of three or more monomeric units. The main monomer groups have been identified in detail, showing a situation very different from that of coal-derived materials, where the sizes of aromatic ring systems increase with molecular size. PMID- 11382932 TI - Simultaneous determination of betamethasone and dexamethasone residues in bovine liver by liquid chromatography/tandem mass spectrometry. AB - In this study a new method for the simultaneous confirmation of betamethasone and dexamethasone residues in bovine liver is presented. A Quattro LCZ triple quadrupole mass spectrometer, equipped with an atmospheric pressure ionization (API) source, was coupled to a high performance liquid chromatograph (HPLC) system. Spiked liver samples were first extracted with acetonitrile, and the extracts were purified on C-18 columns. LC separations were performed on a Hypercarb column, with acetonitrile/water (90:10, v/v, +0.3% formic acid) as the mobile phase. Retention times for dexa- and betamethasone were 6.60 and 8.50 min, respectively. Fluorometholone had a retention time of 6.70 min and was used as the internal standard. The detection of the analytes was performed in the multiple reaction monitoring (MRM) mode. The assay was linear over the range of 0.5 to 8 microg/kg for both analytes. The estimated determination limits were 0.2 microg/kg for both beta- and dexamethasone and the quantification limits were 0.4 microg/kg for dexamethasone and 0.3 microg/kg for betamethasone. Analysis precision at 1, 2 and 4 microg/kg was lower than 6.1% (relative standard deviation, RSD) and accuracy was at least 97.5%. Recoveries at 1, 2 and 4 microg/kg ranged between 56 and 69%. PMID- 11382934 TI - Accuracy and precision of flowing afterglow mass spectrometry for the determination of the deuterium abundance in the headspace of aqueous liquids and exhaled breath water. AB - We have assessed the accuracy and precision of our flowing afterglow mass spectrometric method (FA-MS) for absolute measurements of deuterium abundance in water using standardised tap water/D(2)O mixtures within the D/H range from 155 ppm (local tap water) to 1104 ppm, prepared by the gravimetric method. The results of this study show that a precision and accuracy of 1% can be achieved for the deuterium abundance in water samples. This is quite adequate for the main application, which is the rapid, non-invasive measurement of total body water by deuterium analysis of breath water vapour using the D(2)O dilution method. PMID- 11382933 TI - Analysis of pyridoquinoline derivatives by liquid chromatography/atmospheric pressure chemical ionization mass spectrometry. AB - A method using liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (LC/APCI-MS) has been developed for the characterization and determination of pyridoquinoline derivatives 4,6-bis(dimethylaminoethylamino) 2,8,10-trimethylpyrido[3,2-g]quinoline, 4,6-bis(dimethylaminoethoxy)-2,8,10 trimethylpyrido[3,2-g]quinoline and 4,6-bis[(dimethylaminoethyl)thio]-2,8,10 trimethylpyrido[3,2-g] quinoline, all with potential antitumor properties. LC separation was performed on a conventional C18 column using a binary mobile phase composed of acetonitrile and 50 mM aqueous ammonium formate at pH 3. The APCI mass spectra obtained showed that proton addition giving [M + H]+ was the common mode of ionization to the amino- and thiopyridoquinolines, whereas the alkoxypyridoquinoline was identified by the main formation of the [M - (C2H3)N(CH3)2 + H]+, followed by the [M + H]+ ion. The LC separation conditions and MS detection parameters were optimized for the determination. The analytical method was also applied to the determination of these pyridoquinoline derivatives in fetal calf serum using liquid-liquid extraction with dichloromethane. Acceptable recovery values were obtained, ranging between 45 and 98%. PMID- 11382935 TI - The effect of water on the ion trap analysis of trimethylsilyl derivatives of long-chain fatty acids and alcohols. AB - Gas chromatography/mass spectrometry (GC/MS), with an ion trap mass analyzer, was used to examine the very-long-chain cuticular acid and certain non-acid wax constituents on the leaf sheath surface of Sorghum bicolor before and during 36 hours of light exposure. The mass spectra of the trimethylsilylated acids and alcohols did not match any of those published in searchable mass spectral libraries. The observed differences can be related to the interaction between water and the trimethylsilylated acids and alcohols. Understanding the observed mass spectra of the very-long-chain plant waxes is critical for studies that employ GC/MS with the ion trap mass analyzer to elucidate cuticular wax compositions on plants. PMID- 11382936 TI - In-line coupling of low-pressure short capillary electrochromatography columns to electrospray ionisation mass spectrometry. AB - A new in-house designed and constructed injection valve for capillary electrochromatography (CEC) based on a rotating injection part with compartments for the eluent as well as for the sample has been coupled to a mass spectrometer via a sheath flow electrospray ionisation (ESI) interface, using short capillary columns of 15 cm length. The CEC columns were packed with 3 microm C(18) bonded silica particles, and a mixture of peptides was analysed using an ammonium acetate/acetonitrile eluent. A significant increase in the signal-to-noise ratio was obtained when the peptides were dissolved in water with the same content of organic modifier as in the eluent with an addition of 0.5% (v/v) acetic acid. When the CEC analysis was performed without any additional pressure, the separation current sometimes dropped tremendously due to bubble formation, caused by different permeability in the first and packed part of the column causing an extremely low electroosmotic flow. The separation current was restored to its original value by applying only 7 bar at the inlet of the CEC column, and the separation performance for the test peptides was recovered. A comparison of the CEC performance of peptides in pure CEC mode and in low-pressure CEC mode is reported. PMID- 11382937 TI - Electron ionization mass spectra of bis-1,2,4-oxadiazoles: tandem mass spectrometry and accurate mass measurements. PMID- 11382938 TI - Characterization of stereoisomeric alkaloids by electrospray ionization tandem mass spectrometry. PMID- 11382939 TI - Challenges to interpretation of breast MRI. AB - This review describes the current knowledge and challenges of lesion interpretation with MRI of the breast according to different image interpretation strategies. Particular emphasis is given to patient- and tumor-related factors that influence image interpretation. The impacts of the menstrual cycle, prior surgery, radiation therapy, and chemotherapy are summarized. Particular enhancement features of ductal carcinoma in situ (DCIS) or invasive lobular carcinoma are described. Finally, an adequate diagnosis at MRI of the breast should take into account the results of the patient's history, physical examination, and all imaging tests performed before MRI. J. Magn. Reson. Imaging 2001;13:821-829. PMID- 11382940 TI - Integration of breast MRI in clinical trials. AB - Clinical trials are the mechanism for implementing new treatment methods into clinical practice. A number of organizations have been developed to facilitate the initiation, implementation, and data analysis for clinical trials. Imaging is important for patient selection and establishment of endpoints, yet imaging expertise has been underrepresented in most clinical trial groups. The integration of breast MRI in clinical trials is an important step toward the establishment of this method of imaging in routine practice. In addition, these data will be important in justifying the expense of MRI to payer organizations. The potential roles of breast MRI in clinical trials are reviewed. These breast MRI applications are linked with some of the studies that are under development. It is important for the MRI community to be aware of the importance of clinical trials in the future clinical establishment of breast MRI. J. Magn. Reson. Imaging 2001;13:830-836. PMID- 11382941 TI - MRI: a role in clinical trials. AB - The diagnosis and treatment of breast cancer has evolved significantly over the last 20 years. Breast-conserving therapy is replacing the Halstedian concept of "en bloc" resection. Difficulties in detection, pre- and postoperative planning and follow up continue to challenge the clinician. Women at high risk present a significant clinical dilemma. MRI technology in many of these areas is providing more information about detection, tumor size, extent, and response to treatment. The careful and thoughtful inclusion of MRI in clinical trials may help continue the advancement of breast cancer care. J. Magn. Reson. Imaging 2001;13:837-841. PMID- 11382942 TI - Anatomy and physiology of the female pelvis: MR imaging revisited. AB - This article reviews the normal anatomy of the female pelvis and focuses on uterine physiology, presenting the kinematics of the uterus that can be identified on ultra-fast MR imaging. It also discusses the many facets of the junctional zone on MR imaging. Ultra-fast MR imaging seems to be a powerful tool for evaluating normal anatomy, physiology, and pathology of the uterus. J. Magn. Reson. Imaging 2001;13:842-849. PMID- 11382943 TI - Staging endometrial cancer: role of magnetic resonance imaging. AB - This review article summarizes and comments the role of magnetic resonance imaging (MRI) in the management of endometrial cancer. The MRI technique, appearance, and diagnostic criteria of endometrial carcinoma are discussed. The value of MRI in the preoperative staging of endometrial cancer is compared to alternative strategies. Contrast-enhanced MRI performs best in the pretreatment evaluation of myometrial or cervical invasion, compared to ultrasonography (US), computed tomography (CT), or nonenhanced MRI. The overall costs and accuracy are similar to those of the current methods of staging, including intraoperative gross dissection of the uterus. In addition, results of MRI might decrease the number of unnecessary lymph node dissections. J. Magn. Reson. Imaging 2001;13:850 855. PMID- 11382944 TI - Potential heating effect in the gravid uterus during MR HASTE imaging. AB - Our purpose was to evaluate if temperature changes occur in maternal or fetal tissues during HASTE imaging. METHODS: Pregnant pigs were scanned with the HASTE technique, and temperatures were measured with phase maps and temperature probes inserted into the amniotic fluid and fetal brain. RESULTS: Fiberoptic probes showed that no heating occurred in fetal tissues or amniotic fluid during HASTE imaging. CONCLUSION: Our current HASTE protocols do not deposit a significant amount of heat in the gravid uterus. J. Magn. Reson. Imaging 2001;13:856-861. PMID- 11382945 TI - Critical role of spatial resolution in dynamic contrast-enhanced breast MRI. AB - The spatial resolution of three-dimensional (3D) gradient-echo T1-weighted images, from 40 women with 25 malignant and 23 benign lesions, was purposely degraded to determine the role of spatial resolution in recording, analysis, and diagnosis of dynamic contrast-enhanced breast MRI. Images were recorded and analyzed at pixel resolution according to the 3TP method (Degani et al., Nat Med 1997;3:780-782). Reduction in spatial resolution degraded the appearance of foci with fast wash-in and fast washout dynamics. This resulted in an increase in false-negative diagnoses. The sensitivity for differentiating between malignant and benign lesions, using threshold criteria defined by the 3TP analysis, of 76% decreased to 60% and 24% for a 2- and 4-fold reduction in spatial resolution, respectively, without affecting significantly the high specificity (96-100%). In order to minimize false-negative diagnoses of contrast-enhanced breast MRI and maintain high specificity, it is essential to record and analyze the dynamic behavior at high spatial resolution. J. Magn. Reson. Imaging 2001;13:862-867. PMID- 11382946 TI - MRI vs. histologic measurement of breast cancer following chemotherapy: comparison with x-ray mammography and palpation. AB - Twenty consecutive patients with breast cancer were evaluated following chemotherapy using MRI to assess the size of cancer residua and compare these data with subsequent histologic measurements of the viable tumor. This retrospective study also involved assessment of the preoperative size of the malignancy as determined by physical exam and x-ray mammogram. These values were later compared with the histology. The tumor size correlation coefficient between MRI and pathologic analysis was the highest, at 0.93. Physical exam and x-ray mammography (available for 17 patients) produced correlation coefficients of 0.72 and 0.63, respectively, compared to histologic measurement. The accuracy of MRI did not vary with the size of cancer residua. MRI is an accurate method for preoperative assessment of breast cancer residua following chemotherapy. J. Magn. Reson. Imaging 2001;13:868-875. PMID- 11382947 TI - Texture detection of simulated microcalcification susceptibility effects in magnetic resonance imaging of breasts. AB - The presence, size, structure and clustering characteristics of microcalcifications can indicate breast cancer. The magnetic susceptibility of microcalcifications differs from soft biological tissues, leading to directional blurring effects that can be detected by statistical image processing methods. A study of the ability of statistical texture analysis to detect simulated localized blurring in magnetic resonance imaging (MRI) of dense breast is presented. This method can detect localized blurring with sensitivity of 88.89% to 94.44%, specificity of 99.72% to 100%, positive predictive value of 73.91% to 100% and negative predictive value of 99.91% to 99.95%. J. Magn. Reson. Imaging 2001;13:876-881. PMID- 11382948 TI - Tumor microvascular characterization using ultrasmall superparamagnetic iron oxide particles (USPIO) in an experimental breast cancer model. AB - The diagnostic potential of ultrasmall superparamagnetic iron oxide particles (USPIO) for quantitative tumor microvessel characterization was assessed by kinetic analysis of dynamic magnetic resonance imaging (MRI) in a rodent breast cancer model. Microvascular characteristics (transendothelial permeability (K(PS)) and fractional plasma volume (fPV)) were estimated in 32 female Sprague Dawley rats, bearing breast tumors of varying malignancy. These values were compared to a prototype macromolecular contrast medium standard, albumin (GdDTPA)(30). Transendothelial permeability (K(PS)) correlated significantly (P < 0.05) with the tumor grade (Scarff-Bloom-Richardson (SBR) score) for the USPIO (r = 0.36), as well as for the reference macromolecule, albumin-(GdDTPA)(30) (r = 0.54). Estimates for the fPV did not show a statistically significant correlation with the tumor grade for either contrast medium. In conclusion, USPIO-enhanced MRI data were capable to characterize tumor microvessel properties in this breast cancer model: microvascular permeability (determined using USPIO) correlated significantly with tumor grade. Thus, quantitative estimation of microvascular characteristics in tumors could provide a surrogate of new vessel formation (angiogenesis) and thus a further important clinical indication for USPIO, in addition to MR angiography. J. Magn. Reson. Imaging 2001;13:882-888. PMID- 11382949 TI - Development, standardization, and testing of a lexicon for reporting contrast enhanced breast magnetic resonance imaging studies. AB - The purpose of this study was to develop, standardize, and test reproducibility of a lexicon for reporting contrast-enhanced breast magnetic resonance imaging (MRI) examinations. To standardize breast MRI lesion description and reporting, seven radiologists with extensive breast MRI experience developed consensus on technical detail, clinical history, and terminology reporting to describe kinetic and architectural features of lesions detected on contrast-enhanced breast MR images. This lexicon adapted American College of Radiology Breast Imaging and Data Reporting System terminology for breast MRI reporting, including recommendations for reporting clinical history, technical parameters for breast MRI, descriptions for general breast composition, morphologic and kinetic characteristics of mass lesions or regions of abnormal enhancement, and overall impression and management recommendations. To test morphology reproducibility, seven radiologists assessed morphology characteristics of 85 contrast-enhanced breast MRI studies. Data from each independent reader were used to compute weighted and unweighted kappa (kappa) statistics for interobserver agreement among readers. The MR lexicon differentiates two lesion types, mass and non-mass like enhancement based on morphology and geographical distribution, with descriptors of shape, margin, and internal enhancement. Lexicon testing showed substantial agreement for breast density (kappa = 0.63) and moderate agreement for lesion type (kappa = 0.57), mass margins (kappa = 0.55), and mass shape (kappa = 0.42). Agreement was fair for internal enhancement characteristics. Unweighted kappa statistics showed highest agreement for the terms dense in the breast composition category, mass in lesion type, spiculated and smooth in mass margins, irregular in mass shape, and both dark septations and rim enhancement for internal enhancement characteristics within a mass. The newly developed breast MR lexicon demonstrated moderate interobserver agreement. While breast density and lesion type appear reproducible, other terms require further refinement and testing to lead to a uniform standard language and reporting system for breast MRI. J. Magn. Reson. Imaging 2001;13:889-895. PMID- 11382950 TI - Freehand iMRI-guided large-gauge core needle biopsy: a new minimally invasive technique for diagnosis of enhancing breast lesions. AB - The lack of reliable methods for minimally invasive biopsy of suspicious enhancing breast lesions has hindered the utilization of contrast-enhanced magnetic resonance imaging (MRI) for the detection and diagnosis of breast cancer. In this study, a freehand method was developed for large-gauge core needle biopsy (LCNB) guided by intraprocedural MRI (iMRI). Twenty-seven lesions in nineteen patients were biopsied using iMRI-guided LCNB without significant complications. Diagnostic tissue was obtained in all cases. Nineteen of the 27 lesions were subsequently surgically excised. Histopathologic analysis confirmed that iMRI-guided LCNB correctly distinguished benign lesions from malignancy in 18 of the 19 lesions. The histology revealed by core biopsy was partially discrepant with surgical biopsy in 2 of the other 19 lesions. Freehand iMRI guided LCNB of enhancing breast lesions is promising. Larger studies are needed to determine the smallest lesion that can be sampled reliably and to precisely measure the accuracy of iMRI-guided LCNB as a minimally invasive tool to diagnose suspicious lesions found by breast MRI. J. Magn. Reson. Imaging 2001;13:896-902. PMID- 11382951 TI - Localization and biopsy of breast lesions by magnetic resonance imaging guidance. AB - Contrast-enhanced MRI of the breast has proved to be a valuable tool in the detection and work-up of breast lesions. Most of these lesions are small and not visible by other imaging modalities, such as mammography or US. Thus, only MR guided preoperative localization techniques or MR-guided percutaneous biopsy can provide a histologic work-up of such lesions. MR-guided preoperative localization seems to be a well-established procedure. However, MR-guided biopsy is still problematic. Although prototypical biopsy systems have been developed, considerable progress is still required. Problems exist with MR-guided biopsy due to severe needle artifacts and tissue shift during the intervention. Thus, needle biopsy currently is not recommended for lesions smaller than 10 mm. This work reviews current techniques for MR-guided preoperative localization and percutaneous biopsy in breast lesions. The diagnostic accuracy achievable with these techniques will be discussed, as well as the potential for new research opportunities and directions. J. Magn. Reson. Imaging 2001;13:903-911. PMID- 11382952 TI - Evaluation of female intrapelvic tumors by clinical proton MR spectroscopy. AB - We measured metabolites in large tumors of the female pelvis (23 cases total: 6 malignant cases, 17 benign cases) using single-voxel proton magnetic resonance (MR) spectroscopy and evaluated the clinical significance of this method in the differential diagnosis of female pelvic tumors. The characteristically obtained signal was lactate, which was detected not only in all the malignant tumors but also in some of the benign tumors. However, the lactate signals of the malignant tumors tended to form higher peaks than those of benign tumors, which was a finding consistent with the result of the earlier in vitro study reported in ovarian tumors. The signal of choline-containing compounds (Cho) was found only in solid tumors, and the signal intensity varied among different histological types of tumors, possibly reflecting metabolic activity of the cell membrane. We considered that proton MR spectroscopy (MRS) of intrapelvic tumors would be useful for the diagnosis of the nature of female intrapelvic tumors. J. Magn. Reson. Imaging 2001;13:912-917. PMID- 11382953 TI - Characteristic MR findings of cervical pregnancy. AB - The aim of this study was to define the characteristic MR findings of cervical pregnancy. Twelve patients with cervical pregnancy underwent MRI because of difficulty in the diagnosis by ultrasonography, human chorionic gonadotropin assessment, and other clinical evaluations. The assessment of MRI included size and location of the lesion, margin, MR signal intensity, rim of low-signal intensity, enhancement pattern, appearance of enhancing solid component, parametrial change, endometrial change, pelvic fluid collection, and ovarian change. All cases showed ill-marginated mass with very heterogeneous signal intensity on T2-weighted images, irregular internal high-signal intensities on T1 weighted images, a partial or circumferential rim of low-signal intensity, dense irregular peripheral enhancement and enhancing papillary solid components with accompanying tubular signal voids, and variably increased parametrial vascularities. This heterogeneous hemorrhagic mass with densely enhancing papillary solid components may be the typical MR finding for cervical pregnancy. J. Magn. Reson. Imaging 2001;13:918-922. PMID- 11382954 TI - Dynamic imaging of the pelvic floor using an open-configuration magnetic resonance scanner. AB - The aims of this study were to develop a noninvasive, erect, gravity-dependent method for assessing movements of the female pelvic floor, to describe the range of movements in pelvic floor ascent and descent in asymptomatic and symptomatic women, and to quantify any differences. A total of 102 women, 28-86 years of age, 35 symptomatic and 67 asymptomatic, were included in the study. They were scanned in a sitting position in an open scanner with good vertical access using fast gradient echo sequences fast spoiled grass (FSPGR). Measurements of the bladder base, uterocervical junction, and anorectal junction were taken in the sagittal plane. The levator ani (LA) muscle insertion was assessed in the coronal plane with the patients at rest, during maximal strain, and during maximum contraction of the pelvic floor. Premenopausal multiparous women have a significantly lower bladder base (8/0 mm above the baseline; P value = 0.009) and uterocervical junction (15.5/3.5 mm; P value = 0.03) at rest than nulliparous women, and this becomes more apparent on straining. Parity confers a more significant effect on the position and function of the pelvic floor than menopausal status. All pelvic organs are lower at rest and on straining in women with defecation difficulties (0/-24 mm; P value = 0.001). These differences are also seen when comparing women with and without urinary incontinence. Dynamic seated magnetic resonance imaging (MRI) shows that all the pelvic organs are lower at rest and on straining in multiparous women and in those with urinary incontinence than in a group of asymptomatic nulliparous volunteers. This difference is also seen in the position of the bladder base and anorectal junction during pelvic floor contraction. These findings suggest general pelvic floor weakness in women who present with symptoms in one compartment and indicate the need for evaluation of the entire pelvic floor particularly prior to surgery. J. Magn. Reson. Imaging 2001;13:923-929. PMID- 11382955 TI - T2-weighted MRI of the female pelvis: comparison of breath-hold fast-recovery fast spin-echo and nonbreath-hold fast spin-echo sequences. AB - In 49 patients who had pelvic abnormalities, breath-hold T2-weighted fast recovery (FR)-fast spin-echo (FSE) (imaging time = 24 sec) and nonbreath-hold FSE MR images (2 min 8 sec) were compared qualitatively (on a four-point scale) and quantitatively (using signal-to-noise ratios (SNRs) and contrast ratios (/SIs of the lesions-SIs of the myometrium/SIs of the myometrium)). Motion artifacts were reduced on breath-hold FR-FSE (3.8:3.2 = breath-hold FSE:nonbreath-hold FSE, P < 0.01) and image quality was comparable (3.8:3.7, NS). In all patients, pathology (leiomyoma [N = 26], adenomyosis [N = 10], endometrial carcinoma [N = 8], and ovarian cystic lesions [N = 21]) was recognized with comparable lesion conspicuity (3.8:3.7, NS) and better delineation of the structures (3.9:3.6, P < 0.05) on the FR-FSE images. There was no significant difference in contrast ratios, although SNRs (e.g., myometrium 18.3:25.8, P < 0.01) were better and the uterine zonal anatomy was recognized better on the nonbreath-hold FSE (3.4:3.7, P < 0.05). These differences did not affect the diagnosis. Breath-hold FR-FSE provides the benefits of motionless imaging and a short examination time, although lower SNRs were noted. J. Magn. Reson. Imaging 2001;13:930-937. PMID- 11382956 TI - Fetal shoulder measurements by fast and ultrafast MRI techniques. AB - The purpose of this study was to evaluate magnetic resonance imaging (MRI) of fetal shoulder measurements of fetuses with suspected macrosomia. The actual fetal shoulder measurements made immediately after birth were compared with measurements obtained by fast and ultrafast MRI techniques antepartum. Eight singleton diabetic pregnant mothers underwent MRI examination with fast imaging in steady-state precession (TrueFISP) and spin-echo (SE) and gradient-echo (GE) echo-planar (EPI) sequences to show the fetal shoulder width. The actual shoulder width was measured immediately postpartum by a neonatologist. There was a statistically significant correlation between the MRI measurements and the actual shoulder width (P < 0.001 - P < 0.05) for all sequences. TrueFISP (r = 0.98, P < 0.001) was superior to EPI sequences (r = 0.88, P < 0.01 for SE EPI and r = 0.80, P < 0.05 for GE EPI). The images of all three sequences used were free of major motion artifacts. Fast and ultrafast sequences seem to be reliable for fetal shoulder measurements and the TrueFISP was the most accurate sequence compared to SE and GE echo-planar sequences. J. Magn. Reson. Imaging 2001;13:938-942. PMID- 11382957 TI - Functional MRI with simultaneous EEG recording: feasibility and application to motor and visual activation. AB - The possibility of combining the high spatial resolution of functional magnetic resonance imaging (fMRI) with the high temporal resolution of electroencephalography (EEG) may provide a new tool in cognitive neurophysiology, as well as in clinical applications such as epilepsy. However, the simultaneous recording of EEG and fMRI raises important practical problems: 1) the patients' safety, in particular the risk of skin burns due to electrodes heating; 2) the impairment of the EEG recording by the static magnetic field, as well as by RF and magnetic field gradients used during MRI; and 3) the quality of MR images, which may be affected by the presence of conductors and electronic devices in the MRI bore. Here we present our experiences on 19 normal volunteers who underwent combined fMRI and 16-channel EEG examination. Consistent with previous reports, safety could be assured when performing EEG recordings during fMRI acquisition. Electrophysiological signals recorded with surface EEG were similar inside and outside the 1.5 T magnet. Furthermore, fMRI using motor or visual tasks revealed similar areas of activation when performed with and without 16-channel EEG recording. J. Magn. Reson. Imaging 2001;13:943-948. PMID- 11382958 TI - Line scan diffusion tensor MRI of the cervical spinal cord in preterm infants. AB - Line scan diffusion tensor magenetic resonance imaging (DT-MRI) of the cervical spinal cord was demonstrated in vivo for unsedated preterm (gestational age 24-30 weeks at birth), very low birthweight (birthweight 620-1300 g) infants at postmenstrual ages from 29-40 weeks. Scalar invariant measures of diffusion [apparent diffusion coefficient (ADC) and relative anisotropy (RA)] determined from a cervical cord region of interest in each case are reported, characterizing the maturational status of the normal third trimester and newborn spinal cord. Mean ADC of 11 infants was 1.2 +/- 0.1 microm(2)/msec and the mean RA was 24.3 +/ 4.9%. Normal infant cord neural fiber tract morphology was visualized using a mapping of the predominant diffusion tensor eigenvector. Potential clinical applications of line scan DT-MRI of the spinal cord of preterm and term newborns for assessment of spinal cord injury are discussed. J. Magn. Reson. Imaging 2001;13:949-953. PMID- 11382959 TI - Effect of lung inflation on arterial spin labeling signal in MR perfusion imaging of human lung. AB - The effect of lung inflation on arterial spin-labeling signal in lung perfusion is investigated. Arterial spin-labeling schemes, called alternation of selective inversion pulse (ASI) and its hybrid (HASI), which uses blood water as an endogenous, freely diffusible tracer, were applied to magnetic resonance (MR) perfusion imaging of the lung. Perfusion-weighted images of the lung from nine healthy volunteers were obtained at different time delays. There was a significant signal difference in ASI images acquired at different respiratory phases. Greater signal enhancement has been observed when the volunteers performed breath holding on end expiration than on end inspiration. This is in agreement with the normal physiologic effect of lung inflation on the pressure flow relationship of pulmonary vasculature. ASI and HASI perfusion-weighted images show similar lung features and image quality. Preliminary results from pulmonary embolism patients indicate that arterial spin labeling is sensitive for the detection of areas of perfusion deficit. J. Magn. Reson. Imaging 2001;13:954 959. PMID- 11382960 TI - Fast spin echo and fast gradient echo MRI with low acoustic noise. AB - It has been shown previously that a significant reduction of the acoustic noise in standard, slow magnetic resonance imaging (MRI) sequences is achieved when the gradient pulses have long sinusoidal ramps. An improvement of this method is now presented for fast gradient echo (FLASH) and fast spin echo (RARE) sequences. The new strategy consists of using a sinusoidal readout pulse with no plateau and extending the phase encoding pulses to the entire readout period. The distribution of k-space samples is no longer equidistant, and images have to be reconstructed with gridding. A reduction of the acoustic noise by 20-40 dBA was obtained with respect to standard sequences installed by the scanner manufacturer, which is 10-20 dBA better than that with the previously proposed method of sinusoidal ramps. J. Magn. Reson. Imaging 2001;13:960-966. PMID- 11382961 TI - An integrated visualization system for surgical planning and guidance using image fusion and an open MR. AB - A surgical guidance and visualization system is presented, which uniquely integrates capabilities for data analysis and on-line interventional guidance into the setting of interventional MRI. Various pre-operative scans (T1- and T2 weighted MRI, MR angiography, and functional MRI (fMRI)) are fused and automatically aligned with the operating field of the interventional MR system. Both pre-surgical and intra-operative data may be segmented to generate three dimensional surface models of key anatomical and functional structures. Models are combined in a three-dimensional scene along with reformatted slices that are driven by a tracked surgical device. Thus, pre-operative data augments interventional imaging to expedite tissue characterization and precise localization and targeting. As the surgery progresses, and anatomical changes subsequently reduce the relevance of pre-operative data, interventional data is refreshed for software navigation in true real time. The system has been applied in 45 neurosurgical cases and found to have beneficial utility for planning and guidance. J. Magn. Reson. Imaging 2001;13:967-975. PMID- 11382962 TI - Necropsy by magnetic resonance in a case of conjoined thoracopagus twins. AB - A case of thoracopagus conjoined stillborn twins studied with magnetic resonance imaging (MRI) is presented. Due to the parents' denial of authorization for an autopsy of the fetuses, the MRI study was performed as an alternative to necropsy. High-resolution images of the fetuses demonstrated relevant anatomic features, providing noninvasive pathological insight with preservation of the fetal specimen. J. Magn. Reson. Imaging 2001;13:976-981. PMID- 11382963 TI - Uterine fibroid embolization: assessment of treatment response using perfusion weighted extraslice spin tagging (EST) magnetic resonance imaging. AB - In this pilot study, we demonstrate the feasibility of using an arterial spin tagging technique, Extraslice Spin Tagging (EST), to assess tumor perfusion before and after uterine fibroid embolization (UFE) and correlate the changes in perfusion with fibroid size reduction. We followed two patient volunteers over a six-month period. The perfusion-weighted image intensity decreased immediately after UFE. The size of the tumor decreased by 14% immediately after UFE and continued to decrease over a six-month period to 84%. The imaging methods presented allow for rapid measurement of tumor volume and the evaluation of perfusion of the tumor without the need for intravenous administration of gadolinium compounds. J. Magn. Reson. Imaging 2001;13:982-986. PMID- 11382964 TI - Time course of arterial hyperintensity with fast fluid-attenuated inversion recovery imaging in acute and subacute middle cerebral arterial infarction. AB - The purpose of this study was to evaluate the time course of arterial hyperintensity (AH) in acute and subacute cerebral infarctions of the middle cerebral artery (MCA) distribution by using fast fluid-attenuated inversion recovery (FLAIR) imaging. A total of 40 FLAIR MR examinations were performed in 27 patients with MCA infarction within 13 days after ictus. Thirteen patients underwent two MR examinations during this period. Thrombotic or embolic infarctions were included in this study, but lacunar infarctions were excluded. The presence or absence of AH and the location of AH were analyzed. Overall, AH was found in 24 (60%) of 40 FLAIR examinations within 13 days after onset. AH was seen in 17 (100%) examinations less than 24 hours old, four (40%) of 10 examinations 1-4 days old, two (18%) of 11 examinations 5-9 days old, and one (50%) of two examinations 10-13 days old. AH was most frequently found at the sylvian fissure (87%), followed by the sulci (54%), and the horizontal segment of MCA (29%) in the affected MCA distribution. Although AH could be seen even at 13 days after ictus, the presence of AH declined over time. AH represented an early sign of acute embolic or thrombotic infarction. J. Magn. Reson. Imaging 2001;13:987-990. PMID- 11382966 TI - [Biological basis of orthodontic tooth movement]. AB - The effect of orthodontic therapy is dependent of the biological possibilities and limitations of the dento-alveolar complex. Biomechanical effects determine the first phase of tooth movement. In the second phase hyalinisation occurs in almost all cases. Elimination of the hyalinised tissue is associated with undermining bone resorption. Next, 'real' tooth movement starts. At the pressure side the normal structure of the periodontal ligament is destroyed and so is the tooth attachment. At the tension side deposition of trabecular bone is found and the tooth attachment remains. The regulation of these processes is still not completely understood, but cytokines and growth factors play an important role. The biological system does not react according to a simple dose-response relation and large individual differences in susceptibility of the system exist. PMID- 11382967 TI - [Myth of optimal pressure]. AB - Orthodontic tooth movement always follows the same pattern. Four phases can be distinguished. During the last phase, the linear phase, the tooth moves through the alveolar bone. One could assume that the rate of tooth displacement is related to the magnitude of the force or to the pressure in the periodontal ligament. No consensus exists on the optimal pressure for orthodontic tooth movement. In literature pressures are advocated, ranging from 2 to 30 KPa. Animal experiments show that a large range of force magnitudes results in an equal rate of tooth movement. A dose-response relation is only feasible when forces are used which are far below those used in an everyday practice. PMID- 11382968 TI - [Photography in orthodontics]. AB - To achieve high quality pictures decisions have to be made regarding the camera and lenses, the lighting, the photographic film, the photographic procedures and the registration of the pictures in the files of the patient. The necessity of standardisation and the difficulties in achieving this are mentioned. The use of pictures in orthodontic diagnosis, treatment planning, treatment follow-up and quality control are described too. PMID- 11382969 TI - [Facial analysis without cephalometric radiography]. AB - A careful clinical examination of the face is an essential part of the orthodontic diagnosis. By visual examination of the face important diagnostic determinations can be made regarding the patients profile, dental and skeletal relationships and 'facial animation'. This article presents some clinical and photographic diagnostic methods to analyse the face. The possibilities and limitations of facial analysis without cephalometric radiography are discussed. PMID- 11382970 TI - [Early orthodontic treatment, a diagnostic challenge]. AB - This article investigates which growth anomalies require early orthodontic interception, that is before the age of 9 years. The diagnosis should be established at three levels: the skeletal jaw relationship including the facial profile, dental space problems determined by the apical base area, and function. Rather than to advise on mechanotherapy emphasis is placed on improvement of diagnostic skills because, contrary to therapy, making a diagnosis cannot be delegated. The analysis at the three diagnostic levels shows that only a limited number of situations demand early treatment. Among these are a negative overjet, a cross-bite, open mouth posture, and sometimes a space maintainer after extraction of deciduous molars. Special precautions are required when front teeth become ankylosed at an early age after trauma. In case of doubt the general practitioner is advised to seek orthodontic counseling from a specialist. PMID- 11382971 TI - [Are removable appliances obsolete?]. AB - Removable orthodontic appliances have some limitations compared to fixed appliances. However, a removable appliance can be used during each period of craniofacial growth and dental development. A number of clinical applications for the removable appliance during the mixed and permanent dentition is explained. The use of functional orthopedic appliances and removable retention appliances are also described. It is concluded that the removable appliance can still play a favorable part in the treatment of malocclusions in each developmental stage. PMID- 11382972 TI - [Developments in fixed orthodontic appliances]. AB - The Edgewise system and the Begg technique were until the early 1970's two of the most familiar and well-known fixed appliance techniques in orthodontics. During the following years they both underwent some slight adaptations that were generally based on more straightforward biomechanics. These changes have led to what in recent years is called the 'Straight Wire technique'. The latter claims to be the treatment option of choice based on preadjusted appliances and preformed arch wires. Besides biomechanics, treatment tools also concentrated on esthetics. It resulted in the development of smaller brackets as well as tooth coloured ones made from polycarbonates or ceramics. Finally lingual orthodontics starts to play a more important role in the orthodontic treatment options for the new millennium. PMID- 11382973 TI - [Orthodontic treatment for adults]. AB - The number of adults undergoing orthodontic treatment has increased strongly and the average age that adult patients undergo orthodontic treatment increased steadily although 3/4 is still younger than 27 years. In adults the facial skeletal pattern can only be changed in a very confined way, consequently in case of an abnormal skeletal pattern one has to choose between a combined orthodontic surgical approach (which is the case in 18% of the patients) and a compromised orthodontic treatment, if necessary combined with other disciplines. It is still controversial whether tooth movement in adults is slower and more difficult than in adolescents. The same holds true for the risk for loss of periodontal support, for root resorption, for gnathologic problems and for relapse. As related to these variables there appears to be a large individual variation. Many adults show one or more problems in their dentition that may influence their orthodontic treatment. About 60% of the adult patients need a multidisciplinary approach. The development of implantology and of bone regeneration and bone grafting has lead to more combined treatments. The risks of such complex treatment plans are generally larger than those for more simple kinds of treatment. A very careful treatment planning and good communication between the different specialists is essential. Moreover the treatment plan with all its (dis)advantages has to be extensively discussed with the patient. PMID- 11382974 TI - [Permanent retention: yes or no?]. AB - The (in)stability of orthodontic treatment is a daily problem in orthodontics. This implies that retention--in whatever way--also remains an actual subject. There is however scarce valid scientific evidence on the topic of retention. PMID- 11382975 TI - [Iatrogenic effects of orthodontic therapy]. AB - Iatrogenic effects of orthodontic treatment are root resorption, pulpal changes, decalcifications and white spots, gingival and periodontal changes, enamel surface changes, temporomandibular dysfunction, immunological reactions, pain and discomfort, and accidents. The authors discuss these items and give, if possible, tips to prevent or reduce these effects. PMID- 11382976 TI - [Long-term stability of orthodontic treatment]. AB - Aim of this study was to assess long-term stability of orthodontic treatment in a sample of 1016 patients until 10 years postretention. Treatment outcome was measured with the PAR-index at 6 different stages. The mean age of the patients was 12.0 +/- 3.1 year at the start of treatment to 26.3 +/- 2.9 year 10 years postretention. The results show that 67% of the orthodontic treatment result, as measured with the PAR-index, was maintained 10 years postretention. The PAR scores for the midline and the open bite remained about the same over the years. However, the scores for the lateral occlusion, overjet, reversed overjet, overbite, and contact point displacement of the upper and lower front teeth increased gradually over time. Nearly 50% of the total relapse took place the first two years after retention. The largest change was found for the position of the lower incisors. Ten years postretention their position was even worse than at the start of treatment. PMID- 11382977 TI - [Quality management in orthodontics]. AB - For the Dutch government efficient and effective care aimed at patient satisfaction are key words in the quality policy. Without a structural feedback from patients, the users of care, and the purchasers of care successful development of a quality system is not possible. This is the reason why the Dutch government stimulates coordination and cooperation between three parties involved in health care: providers, purchasers and patients. Three different aspects of quality deserve attention in this respect: professional, relational and organisational quality. The ultimate aim is that every patient in orthodontic treatment can count on care that is based on respect and on treatment in which the discomfort is limited to the minimum. PMID- 11382978 TI - Mediocre media. PMID- 11382979 TI - Scavenging of thorium isotopes in the Arctic regions: implications for the fate of particle-reactive pollutants. AB - The sources of inorganic pollutants to the Arctic areas are reviewed using previously published results. The removal of particle-reactive pollutants is discussed using thorium scavenging as an analog. The scavenging of 234Th from the upper water column (approximately 100 m) and sediment inventory of 230Th from the deep Arctic waters is compared to different ocean basins in the subarctic areas. Such a comparison shows that 234Th is in equilibrium with its parent, 238U, in certain regions of the Canada Basin of the Arctic Ocean, while it is deficient in other regions of the arctic as well as in sub-polar ocean basins. This implies that the particle-reactive pollutants in the deep Arctic of the Canada Basin are less likely to be removed from the deep waters and will eventually be transported out of this area. We have utilized the 230Th inventory in sediments from the Arctic area to determine the removal rates of particle-reactive nuclides. The 230Th inventory in the deep Arctic Ocean of the Canada Basin is much lower than the Norwegian Sea and the Fram Strait of the Arctic as well as all other sub polar world oceans. These observations suggest that any pollutants into the deep Arctic areas of the Canada Basin are less likely to be removed locally and may be transported out of this area. In those areas, the colloidal material could potentially play a major role in the removal of particle-reactive contaminants. PMID- 11382980 TI - Development and validation of biocriteria using benthic macroinvertebrates for North Carolina estuarine waters. AB - Biological criteria were developed to assess water quality in mesohaline and polyhaline estuarine waters in North Carolina. The criteria used a combination of three metrics: a new 'Hilsenhoff-type' biotic index incorporating the sensitivity to pollution of each taxon and its relative abundance, total number of taxa, and number of amphipod and caridean shrimp taxa. Agreement with an independent data set was 90%, as was within site repeatability of this biocriteria over a range of water qualities and salinities. The versatility of the biotic index is discussed relative to other sampling methods and in other parts of the world. PMID- 11382981 TI - Clostridium perfringens as a potential indicator for the presence of sewage solids in marine sediments. AB - Marine sediment cores collected from several depths of water and distances from a California sewage outfall were tested to see if sediments influenced by sewage solids were a reservoir of enteric pathogens, and if concentrations of indicator bacteria were related to the presence of sewage solids. Vertical distributions of microorganisms in marine sediments were determined; there was a decrease of indicator bacteria with increasing sediment depth. Aeromonas was randomly isolated, but none of the enteric bacterial pathogens or viruses were detected. While classic indicator bacteria were of little value in predicting the presence of pathogens, or relative amounts of sewage solids, Clostridium perfringens may be a suitable indicator. Clostridium perfringens concentrations were not related to the presence of pathogens in sediments. PMID- 11382982 TI - Composition, distribution and sources of polycyclic aromatic hydrocarbons in sediments of the Gulf of Trieste, Northern Adriatic Sea. AB - The composition, distribution and the sources of polycyclic aromatic hydrocarbons (PAHs) in the surficial sediments of the Gulf of Trieste were investigated. To document the spatial PAH input, surficial sediment samples from 17 locations throughout the Gulf were analysed. The total PAH load determined in the surficial sediment samples are between 30 and 600 ng g-1, and were the highest in the immediate vicinity of the Port of Trieste. The PAH contents decline rapidly with increasing distance from the shore. The ratios of methylphenanthrenes/phenanthrene and methylpyrene/pyrene are sensitive indicators of the origin of PAH pollution in the Gulf which is mostly pyrolitic. The phenanthrene/anthracene ratio was used to determine the approximate location and distance from the source of PAH pollution, while 1-methy 1-7 isopropylphenanthrene (retene) was used as indicator for forest fires. A sediment depth profile indicates a major increase in the PAH concentrations after the First World War. PMID- 11382983 TI - Holistic environmental assessment and offshore oil field exploration and production. AB - According to UK Government surveys, concern for the environment is growing. Environmental regulation of the industry is becoming wider in its scope and tougher in its implementation. Various techniques are available to assess how the industry can drive down its environmental impact and comply with environmental regulation. Environmental Assessments (EA) required by European law do not cover the whole life cycle of the project that they are analysing. Life Cycle Analysis (LCA) was developed to assess the environmental loadings of a product, process or activity over its entire life cycle. It was the first technique used in environmental analysis that adopted what was described as a holistic approach. It fails this approach by not assessing accidental emissions or environmental impacts other than those that are direct. Cost Benefit Analysis (CBA) offers the opportunity to value environmental effects and appraise a project on the basis of costs and benefits. Not all environmental effects can be valued and of those that can there is considerable uncertainty in their valuation and occurrence. CBA cannot satisfactorily measure the total environmental risk of a project. Consequently there is a need for a technique that overcomes the failures of project-level EA, LCA and CBA, and assesses total environmental risk. Many organizations such as, the British Medical Association, the European Oilfield Speciality Chemicals Association, the Royal Ministry of Petroleum and Energy (Norway) and Shell Expro now recognize that a holistic approach is an integral part of assessing total risk. The Brent SPAR case study highlights the interdisciplinary nature required of any environmental analysis. Holistic Environmental Assessment is recommended as such an environmental analysis. PMID- 11382984 TI - EPA fines university for polluting environment. PMID- 11382985 TI - Changes in macroalgal communities in the vicinity of a Mediterranean sewage outfall after the setting up of a treatment plant. AB - Benthic macroalgal communities of the upper rocky sublittoral were studied in 1995-1996 in the vicinity of the Marseille (Mediterranean, France) sewage outfall, 8 years after the setting up of a wastewater treatment plant and compared to a previous study carried out in 1972-1974. The number of taxa has increased, a clear stational and seasonal gradient of differentiation of the vegetation appeared, and a turf of ephemeral species is taking place of Corallina elongata at sites close to the outfall. These changes may be due to a decrease in pollutant load, the discharge of ferric chlorates used in the treatment process. However, the overall change is much less conspicuous than that described for deeper soft bottom communities, in particular the Cystoseira amantacea community is not still restored. Biological traits of this species (short distance dispersal) and the nature of most pollutants removed from the effluent (solids and organic matter) may explain this phenomenon. PMID- 11382986 TI - Coral bleaching and mortality on artificial and natural reefs in Maldives in 1998, sea surface temperature anomalies and initial recovery. AB - The bleaching and subsequent mortality of branching and massive corals on artificial and natural reefs in the central atolls of Maldives in 1998 are examined with respect to sea surface temperature (SST) anomalies. SST normally peaks in April-May in Maldives. The UK Meteorological Office's Global sea-Ice and SST data set version 2.3 b shows that in 1998 monthly mean SST was 1.2-4 S.D. above the 1950-1999 average during the warmest months (March-June), with the greatest anomaly in May of +2.1 degrees C. Bleaching was first reported in mid April and was severe from late April to mid-May with some recovery evident by late-May. At least 98% of branching corals (Acroporidae, Pocilloporidae) on artificial structures deployed on a reef flat in 1990 died whereas the majority of massive corals (Poritidae, Faviidae, Agariciidae) survived the bleaching. The pre-bleaching coral community on the artificial reefs in 1994 was 95% branching corals and 5% massives (n = 1589); the post-bleaching community was 3% branching corals and 97% massives (n = 248). Significant reductions in live coral cover were seen at all natural reefs surveyed in the central atolls, with average live coral cover decreasing from about 42% to 2%, a 20-fold reduction from pre bleaching levels. A survey of recruitment of juvenile corals to the artificial structures 10 months after the bleaching event showed that 67% of recruits (> or = 0.5 cm diameter) were acroporids and pocilloporids and 33% were from massive families (n = 202) compared to 94% and 6%, respectively, in 1990-1994 (n = 3136). Similar post-bleaching dominance of recruitment by branching corals was seen on nearby natural reef (78% acroporids and pocilloporids; 22% massives). A linear regression of April mean monthly SST against year was highly significant (p < 0.001) and suggests a rise of 0.16 degree C per decade. If this trend continues, by 2030 mean April SST in the central atolls will normally exceed the anomaly level at which corals appear there are susceptible to mass bleaching. PMID- 11382987 TI - Observations on the effects of barite on the gill tissues of the suspension feeder Cerastoderma edule (Linne) and the deposit feeder Macoma balthica (Linne). AB - Barite, an important component of offshore drilling muds, is shown to adversely affect the ctenidia of the suspension feeding bivalve, Cerastoderma edule and the deposit feeder, Macoma balthica. SEM observations showed that exposure to barite caused cilia to shorten and coagulate, and, in some extreme cases, cause the disintegration of the gill structure itself. Using a simple ciliary condition index (CCI) the impact of the barite was quantified and damage rates expressed. The bivalves were treated with daily doses of 1, 2, and 3-mm depth equivalents of barite. In all treatments significant damage to the gills was recorded although, in the case of the 1-mm dose rate, this did not occur for 4 days. In the other two treatments, damage was apparent within a day of exposure with 100% mortality occurring within 12 days. Macoma balthica appeared slightly more tolerant of exposure to barite than C. edule. PMID- 11382988 TI - First National Conference on marine bioinvasions, Massachusetts Institute of Technology, Cambridge, Massachusetts, 24-27 January 1999. PMID- 11382989 TI - The 1999 IAHSS survey--crime in hospitals. AB - The 1999 IAHSS crime survey attracted the largest number of participants since 1991. Their data confirm the continued reported drop in property crimes--fueled by the decline in larceny theft--in the past few surveys. A smaller drop in violent crimes in 1999 compared to the average for the decade is also reported. Most hospital crimes continue to take place inside a building (by three to one) and the hospital employee continues to be the most likely victim. PMID- 11382990 TI - Multiple shifts as a solution to limited resources. AB - How a security manager, faced with a limited budget, was able to find a flexible and cost-effective way to provide security coverage for a large medical campus that includes a trauma center. PMID- 11382991 TI - Training: putting the pieces together. AB - What steps are needed to develop and successfully manage a comprehensive safety and health program for your institution? Don't overlook a long-term action plan and remember that safety is a shared responsibility, says the author. PMID- 11382992 TI - Four methods of training for security officers. AB - Security officers need to maintain and improve their training. The author discusses the various types of training that are available and the merits of each. PMID- 11382993 TI - Tools to cure healthcare ills. AB - How can healthcare staff at diverse and remote off-site locations be protected? The right security technologies balanced with human resources, says the author, can help security professionals find cost-effective ways to handle the difficult challenges of the healthcare environment. PMID- 11382994 TI - Disaster management: the iceman cometh. AB - How a small, rural Canadian hospital learned valuable disaster management lessons when it was called to help the community survive an intense ice storm. PMID- 11382995 TI - Hospital security's two-edged sword: over-reliance on automation, under-reliance on experienced personnel. AB - The article explores the current trend in the healthcare industry of cutting not only personnel--security directors, managers, and security officers--but also training programs in the name of budget cuts. What are the dangers to patient protection? What changes can be made to operate more effectively in this economic climate? Why does over-reliance on automation create problems of its own? PMID- 11382996 TI - Looking for Dr. Jekyll but hiring Mr. Hyde. AB - As healthcare facilities are expected to provide a safe environment for employees and the public, proactive steps must be taken to screen, supervise, and train employees. This article discusses human resource management procedures to avoid liability for negligent hiring, supervision, retention, and training. PMID- 11382997 TI - A healthy hiring approach. AB - The author describes his healthcare system's experience in dealing with a state law requiring healthcare organizations to conduct background checks on potential employees. Security, he says, must play a role in implementing these requirements. PMID- 11382998 TI - Infant kidnapping: a tragic incident provokes a new HCFA response. AB - The death of a newborn successfully kidnapped from a hospital nursery overshadowed a recent record of fewer or no abductions compared to prior years. In this case, for the first time in hundreds of fatal or non-fatal kidnappings, HCFA made serious threats to cut off the hospital's Medicare funding. PMID- 11382999 TI - Baby switching: an underreported problem that needs to be recognized. AB - Baby switching is a problem that has been underreported in the past but that can have the same impact on a healthcare facility as an infant kidnapping. This article discusses incidents of baby switching at various medical centers that have taken place in the past few years and the precautions being taken by those facilities as well as others to prevent such occurrences. PMID- 11383001 TI - What do we gain from collaboration between developing and industrialized countries? PMID- 11383000 TI - Predicting the future of psychiatry. PMID- 11383002 TI - [Mentality and Spirituality of the Japanese]. PMID- 11383004 TI - [Attitude of community psychiatrists toward access to medical records]. PMID- 11383003 TI - [Public music concerts in a psychiatric hospital: effects on public opinion and as therapy for patients]. AB - We investigate the effects of music therapy concerts, which were held 60 times over a four year period, 1992 to 1996, in Geiyo Psychiatric Hospital, Kochi Prefecture and found that; 1) Musicians who performed at the concerts were not only from Kochi prefecture but also from other prefectures (10 times) and from four foreign countries (7 times). 2) Live concerts in a small hall had a positive influence on patients and drew the patient's attention and interest away from their hallucinations and delusions to the real world. Moreover, the concerts provided the patients with chances to acquire social graces such as being well groomed. 3) Explanations by the musicians, interviews with the musicians and the seasonal choruses accompanied by the musicians were helpful to give the patients motives for recovering communication skills and to interact with society. 4) Inquiries to the patients about the concerts indicated discrepancies between the poor observed estimations during the concerts (83.3%) and the good subjective impressions expressed by the patients (82.0%), suggesting that the patients were not good at expressing their internal emotions through facial expressions or attitudes. 5) Many citizens including children came to the concerts and/or gave aid to the hospital because the concerts were open to the public and we suggest that this contributed to improving the general publics' image of psychiatric hospitals. Questionnaires revealed that 90% of people in a control group had a bad image of psychiatric hospitals in Japan, but only 32% of the members of the general public who attended our concerts had a bad image of psychiatric hospitals. In addition, the revolving ratio of the hospital beds rose from 0.4 to 1.2 over the four years, which also suggests a beneficial effect on the patients. PMID- 11383005 TI - [Present status of access to medical records in England--special reference to psychiatry]. PMID- 11383006 TI - [Confidentiality of patient records -- an ethical problem facing the Japanese Society of Psychiatry]. PMID- 11383007 TI - [Conditions for access to medical records in psychiatry viewed from the standpoint of medical education]. PMID- 11383008 TI - [Legal problems in access to medical records in psychiatry]. PMID- 11383009 TI - [Guidelines for the therapy of patients with schizophrenia]. PMID- 11383010 TI - [EBM/algorithm (flow chart) vs. experimental evidence/therapeutic effectiveness- physicians' decision on choice of therapeutic measures]. PMID- 11383011 TI - [Non-McDonaldization of the therapeutic guidelines for mental diseases]. PMID- 11383012 TI - [Current status of autism studies]. AB - The current status of autism studies was reviewed based on English articles published during the 1990s. Although the concepts of autism and pervasive developmental disorders (PDD) are established, diagnostic criteria of PDDNOS or atypical autism, which is frequently difficult to differentiate from autism, need to be established. The prevalence of autism has been estimated as about 0.05% in the U.S and many European countries, while it was reported to be 0.1% or higher in Japan and some European countries, though the reasons for this difference are unclear. High-functioning (IQ > or = 70) autism may not be as rare a condition as previously thought and both its difference from and similarity to Asperger's syndrome, the highest functioning PDD subtype, need clarification. About 20 to 40% of children with autism lose meaningful words by the age of 2 years and display autistic symptoms thereafter. Such autism, called the setback type in Japan, has been demonstrated to have a poorer adolescent/adult outcome compared to autism without setback and its relationship with childhood disintegrative disorder, which displays a clearer regression after normal development for at least the first 2 years of life, needs to be addressed. The etiology of autism is now considered mostly genetic for reasons, such as the significantly higher concordance rate of autism in identical twin pairs (60-80%) than in fraternal twin pairs (0-10%) and an 3-5% incidence of autism among sibs of an autism proband, 30 to 100 times higher than that in the general population. The involvement of several genes is implicated to create susceptibility for autism, yet the responsible genes have not been identified. Although there is no medication to cure autism, some psychotropic drugs, such as antipsychotics and SSRIs, seem effective for behavior problems in autism patients. Psychosocial treatments are the main therapeutic approach to autism, though they are yet to be well systematized. It is important to evaluate various autism treatments now practiced and to integrate effective ones into an efficient therapeutic system. Early diagnosis of PDD, support for the mother to develop a better relationship with her autistic infant, enrollment of an autistic infant in a day care for handicapped or non-handicapped infants, education in a relevant school, vocational training, treatment of behavior problems, and establishing social welfare services are a framework of the therapeutic system of autism. Much should be done to improve the long-term outcome of autism patients, including high functioning autism. PMID- 11383013 TI - [Psychopathological study of twin schizophrenia from the viewpoint of twin interaction]. AB - Schizophrenia research requires both genetical and environmental approaches. Some researchers have studied the environmental aspects of schizophrenia by comparing schizophrenic twins with non-schizophrenic ones in discordant pairs. But few studies have taken notice of the interaction between twins. This article focused on the unique relationship between twins, "inter-twin community", which might be considered one of the most important environmental factors. This study included 14 pairs of twins, 28 people including 17 schizophrenic patients. These inter twin communities were divided into "close type" and "distant type" according to the closeness between the twins prior to the onset of schizophrenia. The close type, in which there was a high degree of closeness, included 8 pairs of twins, 16 people, 11 of whom were patients. The distant type, in which there was a low degree of closeness, included 5 pairs, 10 people, 5 of whom were patients. An additional sub-class of the close type was the "close-competitive type". This sub type had a high degree of closeness and a high degree of competition, and included 1 pair, both of whom were patients. Eight out of the 11 patients in the close type (72.7%) had an excellent prognosis. In this type of inter-twin community the relationships gave patients a strong and stable protective environment. In the close-competitive type, the twins supported each other while one twin was having difficulty, but fought with each other when they recovered and eventually one twin relapsed, resulting in an alternating pattern of schizophrenic episodes. The twins were unable to separate from each other and they both had a poor prognosis; thus their relationship both provided the patients with a degree of protection and had a marked harmful effect. In the distant type, 4 of the 5(80%) patients had an poor prognosis. In this type, when one twin became ill, the other twin did not support them, and the relationship provided little protective effect, resulting in poor outcomes. The findings in this study support the idea that the nature of the inter-twin community has a strong influence on the course of schizophrenia in twins. Additionally, since all the female pairs belonged to the close type, and the male pairs tended to be the distant type, there are probable gender differences in the types of inter-twin communities. Finally, in making therapeutic plans for twin schizophrenics, it is very important to consider their relationship in terms of the inter-twin communities. PMID- 11383014 TI - [Community psychiatric hospital attitude regarding access to health records]. PMID- 11383015 TI - Bold approach curbs antibiotic use for bronchitis. PMID- 11383016 TI - Regular PSA screens undervalued, underutilized in prostate cancer detection. AB - One study after another reports that men aren't getting screened for colorectal cancer and prostate cancer, leading to unnecessary morbidity and mortality--as well as higher costs. Here's what you can do about it. PMID- 11383017 TI - Here's how to shop for in-home DM devices. AB - High-tech home devices like weight scales, glucometers, and peak flow meters that send information to clinicians electronically are becoming increasingly popular disease management tools. But whether or not these gadgets can deliver improved quality of care--and a decent return on investment--depends largely on knowing what to look for when selecting the equipment, and figuring out whether your choice will be easy enough for patients to use. PMID- 11383018 TI - Cross-organizational DM programs gaining ground. AB - Imagine the benefits to patients if health plans and systems created a standard set of preventive care measures, disease management protocols, and patient education materials that hold true throughout a given health care market regardless of what health plan or system a patient is enrolled in. In at least a few cases, this vision of disease management utopia is slowly becoming a reality. PMID- 11383019 TI - Hospital offers free drugs to prevent costly hospitalizations. AB - A financially strapped rural hospital is helping control the costs of indigent care by giving away prescriptions to residents with diabetes, asthma, and other chronic illnesses. The program's small expense is more than offset by the reductions in acute care costs. PMID- 11383020 TI - [Needs for implant therapy in cancer patients; a retrospective study]. AB - After cancer treatment in the head and neck area, mastication and speech are often affected. Some of the problems encountered can be solved by adequate dental rehabilitation. However, dental rehabilitation is often compromised, for various reasons. The change in anatomy due to surgery often results in lack of denture bearing mucosa. The effects of radiotherapy of the salivary glands and the mucosa result in dry oral tissues and diminished retention of removable dentures. Osseointegrated implants can help to solve these problems. Implant treatment has, so far, not been widely used in cancer patients. An analysis was made of 95 consecutive patients with a tumor in the head and neck area. The indication for treatment with osseointegrated implants was reviewed and the need for implants as experienced bij patients was evaluated. Results show that 45% did not need specific prosthetic rehabilitation, and approximately 25% of the patients could benefit from osseointegrated implants. Due to general and local contra indications and patients' refusal, only 3% actually have been treated. For complete oral rehabilitation of this group of patients the use of osseointegrated implants should be considered at an early stage, before the initial tumor treatment. PMID- 11383021 TI - [Restoration materials in overdenture abutment teeth]. AB - To assess the performance of amalgam, resin composite and resin-modified glass ionomer cement as filling material in overdenture abutment teeth, 155 restorations were made to seal the root canal orifices in 49 patients. The three restorative materials were randomly assigned to the abutment teeth using a number of balancing criteria. All patients were reviewed every six months and received the same preventive regimen. Survival was assessed at two levels: complete survival even without maintenance treatments (Scomp) and survival of the original restoration independent from eventual maintenance treatments (Sorig). The calculated overall survival of Sorig and Scomp were 63 +/- 6% and 57 +/- 6% respectively (mean +/- SE). At both levels no statistically significant differences were observed between the survival of the investigated materials. The results of this study did not point out a superior restorative material for the seal of root canal orifices of overdenture abutment teeth. The distribution of failures over the patients indicated a certain patient dependency. PMID- 11383022 TI - [Metronidazole in the treatment of refractory periodontitis]. AB - In this article the clinical and microbiological effects of systemic metronidazole in the treatment of refractory periodontitis in adult patients is discussed. For this study, 27 adult patients with detectable Bacteroides forsythus and culture negative for Actinobacillus actinomycetemcomitans were selected. After renewed supra- and subgingival debridement, patients were treated with systemic metronidazole (500 mg TID) for 7 days. Mean probing pocket depth, probing attachment level and bleeding index significantly improved after metronidazole therapy. Patients that became culture negative for B. forsythus showed a significant better therapy response in comparison to subjects that had detectable B. forsythus post-therapy. The maximum clinical response was observed in patients that became negative for B. forsythus, Porphyromonas gingivalis and Prevotella intermedia after renewed debridement and metronidazole therapy. It is concluded that systemic metronidazole therapy can be effective in refractory adult periodontitis. Microbiological analyses of the subgingival microflora in these patients can assist in selecting patients that may benefit from systemic metronidazole therapy. PMID- 11383023 TI - [Syndromes 22. Rieger's syndrome]. AB - Rieger syndrome is a rare autosomal-dominant disorder characterized by defects of the anterior chamber of the eye, failure of the periumbilical skin to involute, and developmental malformations of the dentition. Recognition of the dental anomalies may result in the early diagnosis of this syndrome and prevent progressive visual loss. PMID- 11383024 TI - [Treatment of an impacted upper cuspid: a different approach]. PMID- 11383025 TI - [Developments in dentistry in the twentieth century 6. Dental radiography]. PMID- 11383026 TI - [Unusual complication of mandibular implantation]. PMID- 11383027 TI - [Gout]. PMID- 11383028 TI - [Human Genome Project on Internet 1]. PMID- 11383029 TI - [Two questions concerning local anesthesia]. PMID- 11383030 TI - [Interface between dentistry and otorhinolaryngology]. AB - In this paper some relevant pathological aspects on the interface of dentistry and ENT surgery are discussed. A major part of the field of ENT surgery is not available for direct inspection and/or evaluation. Therefore, ENT pathology is often overseen or not detected at an early stage. Because of frequent patient dentist contact, also in healthy patients, dentists can play an important role in early detection of ENT pathology. The combination of specific anatomic knowledge of the ENT region and a proper anamnesis are potent tools for a dentist to suspect ENT pathology, even in not easily accessible areas like the hypopharynx and oesophagus. PMID- 11383031 TI - [Treatment protocol for craniomandibular disorder 1. Diagnosis]. AB - In two essays the treatment strategy of the department of Oral Function of the Academic Center of Dentistry Amsterdam for treating craniomandibular disorder (CMD) patients will be presented. This first essay starts with a short description of the symptomatology, classification and etiology of CMD. Then it is described how to diagnose a CMD with the use of a standardized diagnostic protocol. Some important aspects of the differential diagnosis of CMD are discussed. Furthermore it is discussed when a CMD-patient can be treated by the dentist or when referral to a CMD specialist or oral surgeon is necessary. PMID- 11383032 TI - [Developments in dentistry in the twentieth century 7. Caries]. PMID- 11383033 TI - [Actinomycosis of the head and neck region]. PMID- 11383034 TI - [The human genome 1]. PMID- 11383035 TI - [Human Genome Project on Internet 2]. PMID- 11383036 TI - [Prosthetic reconstruction after autotransplantation?]. PMID- 11383037 TI - Management of individual mechanical sewage-treatment systems: how much is needed? AB - Over 2,600 individual mechanical sewage-treatment systems were studied to determine if mandatory maintenance contracts and semiannual inspections with effluent sample collection are adequate to meet discharge standards. The following discharge standards were set: 20 milligrams per liter (mg/L) for five day biochemical oxygen demand (BOD5), 20 mg/L for total suspended solids (TSS), and 400 organisms per 100 milliliters for fecal coliform bacteria. In 1997, 67 percent of the individual mechanical sewage-treatment systems discharging in Will County, Illinois, were in violation of at least one discharge standard. The overall cost of management, shared by the homeowners and the county health department, ranged from $350 to $725 per year. PMID- 11383038 TI - Primary- and secondary-school environmental health science education and the education crisis: a survey of science teachers in Ohio. AB - There is a science education crisis in the United States, with studies showing that U.S. high school graduates are not as well-versed in science as graduates in other countries. Studies also suggest that students are better learners when the environment is used as an integrating theme. Therefore, the time is right to discuss opportunities for integrating environmental health science into kindergarten through 12th grade (K-12) curriculum. The research presented here takes a step toward developing the use of environmental health science as a multidisciplinary theme in the K-12 curriculum. Almost 500 K-12 science teachers in Ohio were surveyed for their opinions about the science education crisis and the role of environmental health science in their current courses of instruction. These teachers had been identified as having an interest in environmental education because of their participation in the Environmental Education Council of Ohio. Nevertheless, the results of the survey suggest that these environmentally oriented science teachers are currently not aware of existing environmental health science learning opportunities. Environmental health practitioners have work to do to educate science teachers about the field and about the ways in which studying environmental health science could alleviate the science education crisis. PMID- 11383039 TI - The KD451 indoor air quality monitor. PMID- 11383040 TI - Are you a supervisor? I'm talking to you! PMID- 11383041 TI - Environmental health and antisocial behavior: implications for public policy. AB - Antisocial behavior persists as a preeminent public policy issue in the United States. A more critical assessment of both the policies and the programs designed to address this problem is necessary, and new risk factors for antisocial behavior need to be investigated. Sufficient evidence exists to hypothesize that exposure to neurotoxins may be a risk factor for antisocial behavior. Neurotoxins such as lead have been demonstrated to affect the cognitive development of children, and impaired cognition is accepted as a risk factor for antisocial behavior. Little consideration has been given, however, to the possible link between neurotoxins and human behavior. This paper presents a biologically and socially plausible justification for this association, reviews the supporting evidence, and emphasizes the need for additional investigation of the phenomenon. Elucidation of this risk factor may lead to new strategies for preventing or mitigating antisocial behaviors among youth and adults. PMID- 11383042 TI - [Pathogenesis and therapy of steroid-induced osteoporosis]. PMID- 11383043 TI - [Significance of cytokines for bone loss--are there possible interventions?]. PMID- 11383044 TI - [Epidemiology of corticosteroid-induced osteoporosis]. PMID- 11383045 TI - [Low dose steroid therapy in rheumatoid arthritis: a model for therapy of inflammatory diseases?--Results of the LDPT Study]. PMID- 11383046 TI - [Pathophysiology of bone loss in rheumatic diseases--do bone markers help in monitoring?]. PMID- 11383047 TI - [Possibilities and indications in osteodensitometry]. PMID- 11383048 TI - [Mechanism of action and effects of glucocorticoids]. PMID- 11383049 TI - Therapy and prophylaxis of corticosteroid induced osteoporosis. PMID- 11383050 TI - Protective effect of folinic acid on low-dose methotrexate genotoxicity. AB - INTRODUCTION: Methotrexate (MTX) is an antineoplastic agent widely used in low dose to treat patients with rheumatoid arthritis (RA). Its side effects can partly be explained by folate antagonism. Folinic acid (Leucovorin) is generally administered with MTX to decrease MTX-induced toxicity. However information regarding the inhibitory effect of folinic acid against cytogenetic damage caused by MTX is limited. The aim of this study was to assess the protective effect of folinic acid against MTX-genotoxicity. METHODS: This study was done on Wistar albino rats and in patients with RA. Forty rats of both sexes were randomized into four equal groups and dosed in the following way: Group-I, distilled water vehicle; Group-II, 0.5 mg/kg folinic acid; Group-III, 0.5 mg/kg MTX; Group-IV, 0.5 mg/kg folinic acid plus 0.5 mg/kg MTX. Doses were given i.p., once daily for 8 consecutive days. A bone marrow chromosomal study and a micronucleus test were performed for each rat. Twenty patients with RA (5 males and 15 females) on a 10 mg weekly dose of MTX, i.m., for one month, were administered the same dose of MTX in addition to 10 mg of folinic acid as a single dose 4 hours after MTX administration, i.m., every week for another 4 weeks. Chromosomal studies as well as a micronucleus test were evaluated for each patient. RESULTS: MTX produced a significant genetic injury as proved by the increased incidence of chromosomal aberration and micronuclei formation in Group-III animals. Inversely, folinic acid (group IV) produced a significant protection against genetic damages induced by MTX. In RA patients, folinic acid provides satisfactory improvement of MTX induced genetic damage. CONCLUSION: Folinic acid has a protective affect against MTX genotoxicity in human as well as in animal models. PMID- 11383051 TI - Loss of functional capacity caused by a delayed onset of DMARD therapy in rheumatoid arthritis. Long-term follow-up results of the Keitel function test. Brief definite report. AB - The aim of this analysis was to investigate the impact of the postponement of DMARD therapy on the functional outcome of rheumatoid arthritis after 10 years of disease. 321 individuals with a disease duration of at least ten years were selected out of a cohort of more than 1800 patients. Two groups were analysed separately: patients who started DMARD therapy within the first year of their disease, and patients who received their first DMARD not earlier than five years after the onset of RA. The Keitel functional index was determined in every patient after 10 years of RA. After 10 years of disease, the swollen joint count and the ESR had decreased in both groups to a comparable degree. However, patients with early treatment performed significantly better in the Keitel test compared to the group with delayed therapy. Although patients with seropositive RA or rheumatoid nodules had a worse outcome in general, the benefit of early treatment was also significant in these subgroups. PMID- 11383052 TI - [Therapy of ankylosing spondylitis (AS) with radium chloride (224SpondylAT)]. PMID- 11383053 TI - [Position of the German Society of Rheumatology on therapy of ankylosing spondylitis (AS) with radium chloride (224SpondylAT)]. PMID- 11383054 TI - [Introduction of the Hannover Competence Center]. PMID- 11383056 TI - [Prognostic factors--for patients and surgeons?]. PMID- 11383055 TI - [Preliminary consensus statement on glucocorticoid-induced osteoporosis]. PMID- 11383057 TI - [Criteria for evaluating prognostic factors]. AB - In oncology prognostic factors are defined as variables (covariates) with independent influence on outcome. Toxicity and complications of treatment are not considered. The significance and independence of a putative prognostic factor has to be proven by multivariate analysis. The results of prognostic factor analysis differ depending on tumor entity and residual tumor status. Sometimes differences are observed for various endpoints and certain patient subgroups. For identification of prognostic factors a systematic step-wise procedure is recommended. A checklist of important methodological aspects is suggested. The acceptance of new prognostic factors requires a standardized way to determine the factors, adequate evidence of significance and independence and the demonstration of additional prognostic information and its clinical relevance. By integration of multiple prognostic factors the present tumor classification will be extended to prognostic systems. PMID- 11383058 TI - [Tumor related prognostic factors--established and hypothetical aspects]. AB - The prognostic assessment of cancer patients is of special importance clinically and scientifically. Prognostic factors are divided into three groups: patient-, treatment- and tumor-related factors. Only a minority of those factors, for example the UICC TNM classification, have been proven by multivariate studies to be of prognostic significance and have an impact on patient management, even then only as prognostic groups. The individual prognosis of a specific patient, which would allow an individualized therapy, cannot yet be adequately assessed. New prognostic factors may result in improved prognostic predictions; currently it is unclear whether this will be accomplished by molecular prognostic parameters assessed on the basis of the primary tumor or rather by the detection of disseminated cancer cells. Of major importance are well-designed clinical studies, in order to prove the clinical benefit of an individualized patient management. PMID- 11383059 TI - [Long-term outcomes in oncological surgery: influence of individual prognostic factors]. AB - In addition to tumor stage there are factors influencing the long-term outcome of an oncological operation. Age per se is not a risk factor; however, comorbidity coinciding with increased age definitely must be regarded as a decisive risk factor. The best prediction of the operative risk is achieved by the ASA classification. Frequently used scores such as ECOG and the Karnofsky index are of little help. Since most elderly patients are excluded from prospective, controlled and randomized studies, only individualized assessments concerning the risk profile are useful at present. However, the overall operative competence of a center is most relevant in the treatment of the so-called "risk patient". PMID- 11383060 TI - [Therapeutic modalities and prognosis]. AB - The individual prognosis of a patient with gastrointestinal cancer is determined by a number of clinical and biological factors. The most relevant prognostic factors are those that can be influenced by the surgeon himself. The completeness of tumor resection, the so-called residual tumor status on the level of the primary tumor in all three dimensions and on the level of the lymphatic drainge is the outstanding factor with an independent influence on the survival of the patient. In addition, the principles of blood-saving preparation with avoidance of blood transfutions, the consideration of no-touch isolation and the complication-free postoperative course have been shown to be independent prognostic factors that can be influenced by the surgeon. There is clear evidence that the hospital volume and the experience of the surgeons, expressed by the number of cases (caseload) in a specific field, has a strong impact on the outcome of a surgical treatment in gastrointestinal cancer patients. To optimize the prognosis of a patient with gastrointestinal cancer one should consider all therapy-related prognostic factors, and therapeutic modalities should be scheduled after a consensus conference (tumor board) of all therapeutic fields involved in the treatment of cancer. PMID- 11383061 TI - [Resection method and functional restoration in soft tissue sarcomas of the extremities]. AB - In soft tissue sarcoma of the extremities wide tumor excision (R0) has become the most important factor for local control. The tactical and technical surgical parameters are defined, which allow for diminished complication rates during tumor biopsy and definite resection. Reconstructive plastic procedures play a key role in coverage of major defects and prevention of problems due to wound infection and irradiation. If localized within the periphery of limbs, the vast majority of cases will require sophisticated techniques to achieve limb salvage. PMID- 11383062 TI - [Plastic surgery reconstruction of the trunk area after resection of malignant tumors and therapy sequelae]. AB - Plastic surgery is playing an increasingly important role in interdisciplinary therapeutic concepts for malignant skin and soft tissue tumors. Immediate or early reconstruction of post-resection defects is important for the patient's rehabilitation. This article describes the principles of surgical oncological concepts for malignant soft tissue tumors of the trunk and gives an overview of the primary and secondary reconstructive options. PMID- 11383063 TI - [Revised approaches in surgery]. PMID- 11383064 TI - [Scientific publication: German, English or both?]. PMID- 11383065 TI - [Role of FDG-PET in oncological surgery]. AB - Fluorine-18 fluorodeoxyglucose positron emission tomography ([18F]FDG-PET) allows in vivo analysis of tissue metabolic activity. Based on the observation that most malignant tumors display a higher metabolic activity than benign tissues, [18F]FDG-PET offers an interesting option for the diagnosis of primary and recurrent malignant tumors. For the oncological surgeon [18F]FDG-PET is particularly helpful for the diagnosis of tumors of the pancreas, colorectum, lung and esophagus. This short review describes the biological basis of [18F]FDG PET and gives a critical discussion of its role in oncological surgery. PMID- 11383066 TI - [PET studies of recurrent rectal carcinoma. Fundamental aspects of lymphatic metastasis of visceral and somatic carcinomas]. AB - We have shown previously that the bodies of vertebrates are made of a somatic and a visceral component, separated by thin fascial sheets (Chirurg 66: 1230). We postulated that advanced visceral cancers are confined in their pattern of lymphatic spread to one or a few distinct metastatic pathways. In this they are fundamentally different from somatic cancers. To investigate this hypothesis in vivo, we evaluated positive lymph nodes (LNs) on PET scan series of 58 consecutive patients with rectum cancers (RC) compared to breast cancers (BC). We conclude that in RC, a visceral cancer spreads mostly along only one caudocranial pathway (regional, para-aortic, mediastinal, supraclavicular LNs) and never affects somatic LNs. In contrast, BC and urogenital carcinomas, as somatic cancers spread to regional somatic LNs, but do not affect visceral LNs. PMID- 11383067 TI - [Resection of colorectal liver metastases. What prognostic factors determine patient selection?]. AB - AIM OF THE STUDY: Based on a consecutive series of patients undergoing liver resection for colorectal metastases, indicators of prognosis and selection criteria were evaluated. PATIENTS AND METHODS: From 1960 to 1998, a total of 654 patients underwent resection of colorectal liver metastases. In 516 patients (78.9%) this was an R0 resection for initial metastatic disease. These patients form the basis for the investigation. RESULTS: 30-day mortality in this group was 5.8%, while the total procedure-related mortality was 8.3%. Significant morbidity was observed in 16% of patients. Follow-up information until 1 January, 2000 was achieved in 99.5% of patients. Including operative mortality, the actuarial 5-, 10-, and 20-year survival is 38 +/- 5%, 27 +/- 6% und 24 +/- 24%, rising to 41 +/ 5%, 29 +/- 6% and 26 +/- 26% after excluding operative deaths. Tumor-free survival is 35 +/- 5% at 5 years. In the multivariate analysis the following factors are associated with decreased crude survival: extrahepatic tumor (P < 0.0001), intraoperative hypotension (P = 0.0001), non-anatomical procedures (P = 0.0002), a metastasis diameter > or = 5 cm (P = 0.0002), unfavourable grading of the primary tumor (P = 0.0003), satellite metastases (P = 0.0069), mesenteric lymph node involvement (P = 0.0260), use of FFP (P = 0.0307) and synchronous diagnosis of metastases (P = 0.1240). With respect to disease-free survival metastasis diameter is first, followed by extrahepatic disease (P < 0.0001 each). Satellite metastases are removed, while the primary tumor site becomes important with inferior results for rectal cancer (P = 0.0188). The other factors remain stable and in the same order. The number of independent tumor nodules as well as the width of resection margin fail to be significant in both univariate and multivariate analysis. CONCLUSION: These results underline the paramount importance of an R0 resection, but diminish the relevance of most commonly used "contraindications". For the actual decision on liver resection, beside the possibility of achieving an R0 situation, safety aspects regarding comorbidity and acceptable extent of parenchyma loss represent the prime limitation. PMID- 11383068 TI - [Prognostic factors for development of peritoneal carcinosis in stomach carcinoma]. AB - OBJECTIVE: Several adjuvant therapy concepts have been developed to improve the treatment of gastric cancer patients. Dealing with intraperitoneal chemotherapy, it seems to be useful to determine suitable prognostic factors for the occurrence of peritoneal carcinosis, as it is possible to select patients who may profit from this therapy. METHODS: Between June 1975 and December 1999 resection of gastric cancer was performed in 575 patients. From 1 January 1986 clinical data were recorded prospectively, before that time, retrospectively. The complete data concerning preoperative diagnosis, operation, histology, postoperative course and survival time were documented in an Excel file for statistical analysis. Multivariate analysis was performed using the chi-squared logistic regression test. RESULTS: Significant correlation was found between the occurrence of peritoneal dissemination and tumour stage pT3, pN2, G3, cancer of the whole stomach and cancer at the anastomotic site after partial gastric resection. Lauren classification, signet-ring cell cancer, liver metastasis and tumour localisation in the distal or proximal stomach showed no significant correlation to peritoneal carcinosis in the multivariate analysis. However, only one-third of patients with liver metastasis had simultaneous peritoneal dissemination. CONCLUSION: The results show a clinical correlation of tumour localisation, infiltration of the serosa, lymph node metastasis and grading with peritoneal dissemination. The present data serve as a basis for further histochemical and molecular biological investigations e.g. of the expression of adhesion molecules to determine the risk of peritoneal tumour dissemination after gastric cancer. PMID- 11383069 TI - [Surgical therapy of alveolar echinococcosis and long-term outcome]. AB - INTRODUCTION: The incidence of alveolar echinococcosis (AE) is low, and studies and progress reports with regard to surgical procedures are rare. METHODS: Retrospective analysis of surgical therapy of AE and its long-term results between 1983 and 2000 by evaluating medical records and questionnaires. SETTING: German university hospital within the endemic area. RESULTS: Twenty-five surgical procedures were performed in 19 patients with AE (12x partial resection of the liver, 3 of them with additional extrahepatic resection; 3x just extrahepatic resection, 4x bilidigestive anastomosis, 5x exploratory laparotomy, 1x bypass procedure). Fifteen patients were operated on the first time with that diagnosis, four due to a relapse. Seven surgical procedures were estimated to be curative, whereas 18 were palliative, because the parasitic mass could not be resected in toto. One patient died from persistent systemic sepsis as a consequence of microbial superinvasion of a splenic parasitic mass. Morbidity was 28%. All patients had additional medical treatment and periodic follow-up. Three of seven patients estimated for curative surgery developed a relapse. One of the patients discharged following palliative surgery died 13 years after diagnosis with liver insufficiency. Advances in conservative and interventional treatments have greatly improved the prognosis of the disease. CONCLUSION: Curative surgery for AE is feasible only in a minority of patients, because frequently the disease has already spread widely when diagnosed. The minimum distance between the lesion and the cut surface should be 2 cm. Taking the advances in conservative treatment into consideration, the benefit of palliative surgery is uncertain and today there is no evidence for prolonged survival by palliative surgical procedures. Palliative surgery should therefore be reserved for cases with complications that could not be managed by conservative and interventional treatment. PMID- 11383070 TI - [Perianal mucinous adenocarcinoma. A further reason for histological study of anal fistula or anorectal abscess]. AB - A 39-year-old man came to us for surgical treatment of a hidradenitis suppurativa. Upon excision of a perianal abscess, the diagnosis of a rare tumor, a perianal mucinous adenocarcinoma (pT4, pN 1, MO), was made. An abdominoperineal resection was performed, followed by a combination of adjuvant radiation and chemotherapy. A year after the operation, the patient is doing well without any signs of recurrence. This carcinoma probably arises in the anal glands. It often presents as a perirectal abscess and/or an anal fistula. Therefore, the diagnosis is often delayed. At presentation, the tumor is bigger than 5 cm in diameter in 80% of the cases, and the prognosis is poor. It metastasizes mostly to the superficial inguinal or to the retrorectal lymph nodes. There are only case reports and no comparative studies in the literature. In the last 10 years, the carcinoma has mostly been treated by neoadjuvant radiation and chemotherapy, followed by abdominoperineal resection. Since then, the median survival has increased to 3 years. This is the first case report of a combination of a perianal mucinous adenocarcinoma with a hidradenitis suppurativa. PMID- 11383071 TI - [Intraoperative monitoring of intact parathyroid hormone (iPTH) in surgery of primary hyperparathyroidism with a new rapid test]. AB - OBJECTIVES: Intraoperative differentiation between adenoma and hyperplasia during surgery for primary hyperparathyroidism (pHPT) is sometimes difficult, but essential for good results. The aim of our study was to evaluate a new highly sensitive electrochemiluminescence immunoassay (ECLIA) for intraoperative monitoring of intact parathyroid hormone (iPTH) following parathyroidectomy as an adjunct for identification of solitary adenoma in patients with pHPT. METHODS: Thirty consecutive patients with pHPT (2 with recurrent pHPT) were examined following a standardized protocol: Immediately before and 5, 10 and 15 min following parathyroidectomy of the enlarged gland, iPTH was measured with a new ECLIA (Roche-Diagnostics, Mannheim, Germany). The results were available within 15-20 min. Besides 20 conventional bilateral neck explorations, parathyroidectomy was carried out in a minimally invasive video-assisted technique (MI-VAP) in 10 patients. RESULTS: Among the 30 patients we found 24 with solitary adenoma (80%), 5 with hyperplasia (17%) and one with a double adenoma (3%). Five minutes after removal of a solitary adenoma the level of iPTH had decreased by 65 (12)% [mean (+/- SD)], after 10 min by 76 (8)% and after 15 min by 81 (8)%. All patients with multiple gland disease could be clearly identified, as iPTH after 15 min did not fall below 50% of basal value. Only after removal of all hyperplastic glands did iPTH decrease to the normal range. Sensitivity and specificity for prediction of a solitary adenoma were 92% and 100% (decline of iPTH more than 50% from baseline value 5 min after parathyroidectomy). In one patient with recurrent pHPT intraoperative sampling from different sites in both internal jugular veins could predict the quadrant of the enlarged gland. Correlation (r) between the results of the quick and the conventional assay, which requires 24 h of incubation, was 0.955. All patients had normal or low calcium levels postoperatively. CONCLUSIONS: (1) Intraoperative monitoring of iPTH with this new quick assay allows safe identification of patients with solitary adenoma during surgery for pHPT. (2) It represents a valuable adjunct to surgical skill not only in primary operations for pHPT but especially in cases of recurrent surgery for pHPT. (3) With this test available minimally invasive techniques for parathyroidectomy may be employed in cases of preoperatively localized adenoma (ultrasound, sesta-mibi scan), avoiding bilateral neck exploration with its higher potential for complications. PMID- 11383072 TI - [Endobronchial juvenile hemangioma in infancy--removal by main bronchus cuff resection. Case report]. AB - The rare case of a 3.5 month-old infant diagnosed with a endobronchial haemangioma is presented and discussed against a background of reference literary material. A main-bronchus sleeve resection with end-to-end anastomosis was performed without reduction of lung parenchyma. No complications were documented during the period of observation. The results of repeated bronchoscopy showed the post-operative period to be of a non-problematic nature. PMID- 11383073 TI - [Hand-assisted, laparoscopic nephrectomy for live donor organ harvesting. Initial results]. AB - We report technique and preliminary results with a new, laparoscopic hand assisted, technique for live donor nephrectomy. A special device (Pneumo-Sleeve, Handport) in a short (7.5-8 cm) Pfannenstiel incision allows the surgeon to insert his or her hand into the insufflated abdominal cavity and to use it as a "multifunctional instrument" while pneumoperitoneum is preserved. This technique offers significant advantages over the pure laparoscopic approach in terms of maintained tactile feedback, facilitated exposure and dissection, decreased operation time and easy retrieval of specimen with minimized allograft warm ischemia time. PMID- 11383074 TI - [C-arch navigation for transoral atlanto-axial screw placement]. PMID- 11383075 TI - [Gastrointestinal autonomic nerve tumor (GANT)--a rate tumor of the ileum]. AB - The gastrointestinal autonomic nerve tumor (GANT) is an uncommon stromal tumor of the intestinal tract and retroperitoneum first described by Herrera and associates in 1984. GAN tumors, also termed "plexosarcomas", arise from autonomic nervous system plexuses of the gastrointestinal tract. We report a case of GAN tumor of the intestinal tract in a 63-year-old woman. The diagnosis is based on light microscopy and immunohistochemical analyses. The tumor stained positive for neuron-specific enolase (NSE) and S-100 protein and was negative for muscle markers. Pain and chronic and acute bleeding are the most frequent but not specific symptoms, and the diagnostic delay is reflected by a large diameter of these tumors. GAN tumors are fatal and must be considered malignant. They need radical surgical resection. PMID- 11383076 TI - [Isolated complete duodenal avulsion at the pylorus after blunt abdominal trauma]. AB - We would like to place on record a case of an isolated complete duodenal avulsion distal to the pylorus after blunt abdominal trauma, in a 78-year-old patient. From all relevant literature, this is the first case known to us of duodenal rupture, around its complete circumference, in close proximity to the pylorus, which did not result in the injury of any further organs, in the absence of chronically infectious alterations. A duodenal injury in the case of blunt abdominal trauma is rare and amongst injuries to hollow organs is of no great importance. In isolated cases of duodenal injuries, it is normally the second or third section that is affected, both of which are fairly rigidly anchored to the retroperitoneum. If the first section is injured this is normally accompanied by considerable injury to the liver or head of the pancreas. PMID- 11383077 TI - [Repeat rupture of covered rupture of non-anastomotic false aneurysm after femoropopliteal bypass]. AB - For arterial reconstructions veins or alloplastic vascular prostheses are used. Nonanastomotic pseudoaneurysms of vascular protheses are a rare event. Breaks of the suture material, structural defects of the transplant fabric or degenerative modifications in the recipient vessel can cause nonanastomotic or anastomotic pseudoaneurysms. Such pseudoaneurysms are known from early clinical application of ePTFE grafts. In the case presented a nonanastomotic rupture of a ePTFE graft is described with partial repair of the rupture site by fibroblasts and connective tissue, which prevented bleeding. PMID- 11383078 TI - [Cystic space-occupying lesion with involvement of the pelvis: unusual site of echinococcosis. Case report and surgical treatment]. AB - A case of an unusual localization of cystic echinococcosis is presented. Initially it appeared as a retropertioneal tumor with bone involvement and the diagnosis was only recognized intraoperatively. The patient was treated by radical surgery and chemotherapy according to the guidelines of the WHO. We conclude that echinococcosis should be considered in extrahepatic cystic tumors, especially in patients from endemic areas. PMID- 11383079 TI - [Teaching and learning in surgery--The Gottingen curriculum]. AB - The study of medicine in Germany is in need of reform. Oversubscribed courses, the lack of practical reference, scarce patient contact and cancelled lessons define the educational landscape for many students. Since at present we cannot carry out global reforms, the quality of medical education is highly dependent on the commitment of those responsible in the individual institution. The aim of the revised surgical curriculum in Gottingen is to demonstrate how medical education with a high didactic level including multimedia teaching can be realised despite large student numbers. Duties both in patient care and education are coordinated through integrated organisation. Educational content and structure are made transparent by online guidebooks. In the surgical examination course and the practical course in surgery, patient-oriented tuition takes place consistently in small groups. The Teaching Studio provides additional aids such as interactive CD ROMs, online research, textbooks, ultrasound equipment, examination and suturing dummies. Curriculum-relevant information, including the weekly case examples and instruction guides for clinical practical experience, is presented on an Internet website. Constant evaluation of the curriculum not only documents student motivation, but also serves to improve the educational concepts on a continuous basis. On completion of the practical course in surgery, 65% of participants maintain that they have reached the defined learning objectives "well" or "very well" (prior to reform only 17%). In the summer semester 2000, 46% of students gave top marks in the practical course in surgery with respect to tuition in patient contact. The results of the evaluations verify the positive feedback from the student body. PMID- 11383080 TI - [How are beginning students in surgery selected? Results of a survey of German consultant surgeons]. AB - AIM: After finishing medical school it is getting more and more difficult for medical students to get one of the rare positions in a surgical training program. Only a good career plan and an intelligent strategy for the first steps in surgical life seem to be an advantage for that competition. On the other hand the selection of basic surgical trainees only based on the curriculum vitae and brief interviews may not be satisfying. The aim of this investigation was to answer the following questions: 1. How do consultant surgeons select their basic trainees? 2. How important is the applicant's record and the interview? 3. Which educational and personal criteria are important for the selection? 4. How can medical students improve their chance of getting a position in a surgical training program. METHODS: Using an anonymous and standardized questionnaire 39 chiefs of university departments of surgery and 44 consultant surgeons each from general and local hospitals were interviewed concerning their selection process and criteria required for application in their training program. The candidates were chosen from an official address list (Deutsche Chirurgie 98, Berufsverband der Deutschen Chirurgen). The questionnaire was sent to the candidates by official postal service together with a prepaid return envelope. RESULTS: Twenty eight chiefs of university departments of surgery (71.8%) and 29 (65.9%) and 20 (45.5%) of the interviewed consultant surgeons in general and local hospitals responded to the 89-item questionnaire. It turned out that the majority of applications were send to the clinics at the applicant's initiation. Personal relationships or networks are important. Position advertisements are not important. 85.1% of the responders are satisfied with their procedure of selection, nevertheless, 43.4% are interested in recruiting markets. In detail, chiefs of surgical departments in university clinics get a mean of 12.3 (5-50) applications for basic surgical training per month, consultant surgeons in general hospitals 6.5 (4-20), those in local hospitals 5.6 (2-10). The completion and the contents of the doctor's thesis, former research efforts, foreign languages, computer knowledge and other qualifications are the most important criteria in the selection process. Reliability, loyalty, physical and psychological stability, organising and planning ability, stamina, sociability and independence are the most valued personal skills. CONCLUSIONS: Despite the satisfaction with the established common selection process, the high rate of participation in our study shows how important this subject is ranked by leading surgeons. It turned out that besides professional qualification, personal skills are a major criterion in the selection of basic surgical trainees. PMID- 11383081 TI - [Comment on Ch. Nies et al.: Outcome after minimally invasive surgery- qualitative analysis and evaluation of clinical relevance of study points by patient and physician]. PMID- 11383082 TI - [Comment on U. Steinhilper et al.: Micro-invasive laparoscopic cholecystectomy with 2 mm instruments. Introduction of the method and initial results]]. PMID- 11383083 TI - [Varicose vein operations with tumescence local anesthesia--an effective method]. PMID- 11383084 TI - [Diagnosis and therapy of bone metastases]. PMID- 11383086 TI - [DRG practice]. PMID- 11383085 TI - [Comment of the DRG committe]. PMID- 11383087 TI - [Structural change in inpatient management as a challenge for surgery]. PMID- 11383089 TI - [Which private health insurance carriers promote ambulatory surgery?]. PMID- 11383090 TI - [Sick competitors. The federal administration must decide how much competition there will be in health care]. PMID- 11383091 TI - [Resterilization and reuse of a femoral nail]. PMID- 11383093 TI - A new deal in the drug war, sort of. An emphasis on treatment--and the bad guys. PMID- 11383094 TI - In the DNA vials, secrets to steal. For more companies, spying is the easy R & D. PMID- 11383095 TI - Cancer's mechanics. Targeting errant cells. PMID- 11383096 TI - Growing old in a good-home. PMID- 11383097 TI - The assisted-living dilemma. Residents and regulators strain to balance freedom with protection. PMID- 11383098 TI - Box jellyfish (Carybdea alata) in Waikiki: their influx cycle plus the analgesic effect of hot and cold packs on their stings to swimmers at the beach: a randomized, placebo-controlled, clinical trial. AB - The study measured the analgesic effect of hot and cold packs on box jellyfish (Carybdea alata) stings to Waikiki swimmers at the beach. Analysis of data showed a minimal trend toward pain relief 10 minutes after the application of hot packs, particularly when the initial pain was mild to moderate. Cold packs showed no clinically significant relief of pain, compared to the control. Data tracking shows that most box jellyfish appear in Waikiki waters on the 9th or 10th day after the full moon. PMID- 11383099 TI - Case report. Pseudo pseudothrombocytopenia. AB - An 82 year old Caucasian male minister presented with thrombocytopenia and platelet clumping on the blood smear thought to have pseudo thrombocytopenia. However, the platelet clumping was due to delay in making the blood smear rather than clumping due to EDTA anticoagulant. This factitious or artificial platelet clumping (pseudo pseudothrombocytopenia) should not be confused with pseudo thrombocytopenia. PMID- 11383100 TI - Indigenous ways of healing guinea worm by the Sonninke culture in Mauritania, West Africa. PMID- 11383101 TI - Risk perception, future land use and stewardship: comparison of attitudes about Hanford Site and Idaho National Engineering and Environmental Laboratory. AB - With the ending of the Cold War, the Department of Energy (DOE) is evaluating mission, future land use and stewardship of departmental facilities. This paper compares the environmental concerns and future use preferences of 351 people interviewed at Lewiston, Idaho, about the Hanford Site and Idaho National Engineering and Environmental Laboratory (INEEL), two of DOE's largest sites. Although most subjects lived closer to Hanford than INEEL, most resided in the same state as INEEL. Therefore their economic interests might be more closely allied with INEEL, while their health concerns might be more related to Hanford. Few lived close enough to either site to be directly affected economically. We test the null hypotheses that there are no differences in environmental concerns and future land-use preferences as a function of DOE site, sex, age and education. When asked to list their major concerns about the sites, more people listed human health and safety, and environmental concerns about Hanford compared to INEEL. When asked to list their preferred future land uses, 49% of subjects did not have any for INEEL, whereas only 35% did not know for Hanford. The highest preferred land uses for both sites were as a National Environmental Research Park (NERP), and for camping, hunting, hiking, and fishing. Except for returning the land to the tribes and increased nuclear storage, subjects rated all future uses as more preferred at INEEL than Hanford. Taken together, these data suggest that the people interviewed know more about Hanford, are more concerned about Hanford, rate recreational uses and NERP as their highest preferred land use, and feel that INEEL is more suited for most land uses than Handford. Overall rankings for future land uses were remarkably similar between the sites, indicating that for these stakeholders, DOE lands should be preserved for research and recreation. These preferences should be taken into account when planning for long-term stewardship at these two DOE sites. PMID- 11383102 TI - Industrial-waste management in developing countries: the case of Lebanon. AB - This paper presents a critical assessment of the existing Lebanese industrial sector, namely the current status and classification of industrial establishments based on a comparative synthesis and analysis of recent nationwide surveys and studies pertaining to industrial-waste management. Characterisation of solid and liquid industrial wastes generated, including hazardous wastes, is presented together with current and projected waste loads, recycling opportunities, and export/import practices. Institutional capacity and needs pertaining to the enforcement of relevant environmental legislation, staffing and resources, monitoring schemes, and public participation are critically evaluated. Finally, realistic options for industrial-waste management in the context of country specific institutional economic and technical limitations are outlined. The industrial sector in Lebanon consists of small-scale industries (84% employ less than 10 persons), primarily involved in light manufacturing (96%). These industries which are distributed among 41 ill-defined zones and deficient in appropriate physical infrastructure, generate solid, liquid, and hazardous waste estimated at 346,730 tons/year, 20,169,600 m3/year and between 3000 to 15,000 tons/year, respectively. Although the growth of this sector contributes significantly to the socio-economic development of the country (industry accounts for 17% of the gross domestic product), in the absence of a comprehensive environmental management plan, this expansion may not be sustained into the coming millennium. The anticipated expansion will inevitably amplify adverse environmental impacts associated with industrial activities due to rising waste volumes and improper waste handling and disposal practices. These impacts are further aggravated by a deficient institutional framework, a lack of adequate environmental laws, and lax enforcement of regulations governing industrial-waste management. PMID- 11383103 TI - Farmers' involvement in landscape activities: an analysis of the relationship between farm location, farm characteristics and landscape changes in two study areas in Jutland, Denmark. AB - Farmers are important agents in rural landscape management as they modify landscape elements to suit their needs. The purpose of this study is to investigate if patterns of landscape activities undertaken by farmers are related to certain farm characteristics and if landscape activities vary from one location to another. With this purpose, we investigated the differences between the level of farmers' involvement in landscape activities in two study areas in central Jutland, Denmark. Particular attention was given to the analysis of the type and extent of landscape activities and their relationship with farm characteristics. In both study areas, landscape activities leading to a more extensive type of land use predominate. However, a multivariate analysis implemented on the set of landscape activity data reveals significant differences between the two study areas. Hedgerow planting and removal is more common in Sonder Omme, while pond digging and cultivation of permanent grass lands are characteristic activities in Gadbjerg-Givskud. Moreover, the proportion of farmers involved and the area affected by different landscape activities is larger in Sonder Omme than in Gadbjerg-Givskud. The farms of the two areas are different in terms of production type, size and socio-economic characteristics. These characteristics represent significant relationships with landscape activities. However, they are not sufficient to explain the differences between landscape activities undertaken in the two areas. The differences are caused by unique cultural and biophysical environment, which influence and modify the relationship between farm characteristics and landscape activities in a distinct manner for each study area. PMID- 11383104 TI - Sustainable wastewater management for small communities in the Middle East and North Africa. AB - Accelerated expansion of wastewater services to small communities in the Middle East and North Africa (MENA) is essential in order to address serious concerns over water scarcity and pollution in addition to meeting the demand for convenience and protecting public health. Centralized and conventional wastewater systems are currently the preferred choice of planners and decision-makers in MENA. Water and funding are not available to provide these centralized conventional services to small communities. This paper presents an integrated approach to sustainable wastewater management for small communities in MENA under the severe water resources crisis. The approach calls for a paradigm shift from centralized conventional wastewater systems to decentralized wastewater systems. Management of wastewater in MENA should start at home. Wastewater generation should be reduced through a combination of domestic water conservation measures. On-site systems must be improved and monitored to control pollution and to recover water for non-potable water uses. Should the circumstances not allow the use of on-site systems, wastewater should be transported and managed through a community system applying the principles of decentralized wastewater management and using the settled sewers for wastewater transportation where appropriate. This approach will facilitate the accelerated and sustainable extension of environmentally responsible wastewater services to MENA's small communities. It offers great potential for cost reduction, accommodates the necessary domestic water conservation efforts, reduces freshwater inputs in wastewater transportation thus eliminating unnecessary demand on freshwater, reduces associated environmental risks and increases wastewater reuse opportunities. PMID- 11383105 TI - Conservation strategies for effective land management of protected areas using an erosion prediction information system (EPIS). AB - This research demonstrates the predictive modeling capabilities of a geographic information system (GIS)-based soil erosion potential model to assess the effects of implementing land use change within a tropical watershed. The Revised Universal Soil Loss Equation (RUSLE) was integrated with a GIS to produce an Erosion Prediction Information System (EPIS) and modified to reflect conditions found in the mountainous tropics. Research was conducted in the Zenzontia subcatchment of the Rio Ayuquila, located within the Sierra de Manantlan Biosphere Reserve (SMBR), Mexico. Expanding agricultural activities within this area will accentuate the already high rate of soil erosion and resultant sediment loading occurring in the Rio Ayuquila. Two land-use change scenarios are modeled with the EPIS: (1) implementation of soil conservation practices in erosion prone locations; and (2) selection of sites for agricultural expansion which minimize potential soil loss. Confronted with limited financial resources and the necessity for expedient action, managers of the SMBR can draw upon the predictive capacity of the EPIS to facilitate rapid and informed land-use planning decisions. PMID- 11383106 TI - Efficient regional ozone control strategies for the eastern United States. AB - When environmental regulatory bodies formulate control plans, it is incumbent upon them to try to achieve the stated goals in an economically efficient manner. The US Environmental Protection Agency (EPA) is presently developing regulations to limit the influence of transported ozone on areas that are having difficulty meeting the ambient air quality standard. EPA has proposed stringent control measures for emissions of nitrogen oxides (NOx) in 22 states of the eastern US. The strategy would necessitate the use of selective catalytic reduction or similar high-performance technology on almost all major power plants in the region, as well as extensive controls on industrial sources. This paper suggests several alternative approaches that would achieve equal or better environmental improvement at lower cost. These include focusing control efforts on sources closer to the North-east Corridor, pushing controls on close-in sources to a higher level of technology performance, and relaxing the stringency of requirements for states remote from ozone problem areas. All the approaches examined are two to three times more cost-effective than EPA's proposed approach in the North-east Corridor. PMID- 11383107 TI - Targeting sites for conservation: using a patch-based ranking scheme to assess conservation potential. AB - Rare habitats are increasingly threatened by fragmentation and measures are required to conserve these valuable resources. Here, we present a method of targeting habitat patches for conservation using a Geographical Information System. We ranked patches of chalk grassland in the Chiltern Hills Area of Outstanding Natural Beauty using simple abiotic criteria, namely: patch area, patch shape, proximity to areas of the same habitat and surrounding land-use type. We compiled a regionally specific list of indicator species and ranked the habitat patches based on their alpha diversity (calculated from species richness). We compared the results of the two ranking schemes and identified key aggregations of the existing reserve network. These could form the basis of future habitat expansion as required by the United Kingdom Biodiversity Action Plan. PMID- 11383108 TI - Estimating Markov transitions. PMID- 11383109 TI - RX for nosebleed prices. PMID- 11383110 TI - Closing in on cancer. PMID- 11383111 TI - Can gays switch sides? PMID- 11383112 TI - Ah, the blue smell of it! PMID- 11383113 TI - Scourge of the playground. PMID- 11383114 TI - Close encounters. PMID- 11383115 TI - Can the juice! If your kids drink too much sweet stuff, they will miss essential nutrients--and put on pounds. PMID- 11383116 TI - [Vestibular toxicity of gentamycin in newborn infants]. AB - BACKGROUND: Gentamicin known as ototoxic drug is routinely used for treatment of vital infections in neonatologic departments. The aim of the present study is to clarify whether gentamicin has a vestibulotoxic effect when used in therapeutic levels in the newborn phase. METHOD: Children were taken on the knees of the mother or relative sitting on an electronically driven rotating chair. The rotatory stimulation consisted of an undamped sinusoidal stimulus pattern which was performed in total darkness. Horizontal and vertical eye movements were recorded electronystagmographically. PATIENTS: The patient group consisted of 30 children aged between 3.1 and 32.9 months. These children had been treated with gentamicin during the newborn period. A group of 30 healthy children of similar age without gentamicin treatment was the control group. RESULTS: The statistical means of the nystagmus reactions during sinusoidal rotation were similar in both groups. No increase of spontaneous eye movements was seen in the patients or in the control group. CONCLUSIONS: Gentamicin is an important antibiotic for treatment of life-threatening infectious diseases, which acts less ototoxically in controlled therapeutic doses in newborns than in later childhood or in adults. PMID- 11383117 TI - [Magnetic resonance tomography imaging of the inner ear of patients with sensorineural hearing loss or vertigo]. AB - BACKGROUND: Progress in surgical techniques in the temporal bone has lead to the demand of exact delineation of this complex anatomical region for preoperative planning. The aim of this study was to assess the potential of a three dimensional T2 weighted turbo-spin-echo-sequence to depict anatomical details and pathological changes of the inner ear. METHODS: Twenty-five patients presenting with sensori-neural hearing loss and/or vertigo were included in this study. A T2 weighted turbo-spin-echo-sequence was carried out on a 1.5 T imager, employing a surface coil with a diameter of 8 cm. The scan parameters were set as follows: TR 2000, TE 500, scan matrix 128 x 128, field of view 90 mm, slice thickness 0.66 mm, NSA 4. In addition, 3D maximum-intensity-projections (MIP) were created, using these data as source images. RESULTS: Image quality was excellent in all 25 cases. The 2.5 cochlear windings, modiolus, vestibule with saccule and utricle all three semicircular canals, crus commune and the facial nerve and vestibulocochlear nerve in the inner auditory canal were clearly delineated in all patients. The cochlear aqueduct was seen in 32% (8 cases), while the vestibular aqueduct was delineated in 16% (4 cases) in this series. In MIP, the entire labyrinth was shown in a single view. In 22 patients regular morphology was found, while in three cases pathological changes were detected (an enlarged vestibular aqueduct and saccus in one patient, partial fibrous obliteration of the labyrinth in two patients). CONCLUSIONS: The introduced TSE sequence can be used to delineate the anatomy of the inner ear as well as pathological changes. The technique appears to be useful for preoperative planning. PMID- 11383118 TI - [Detection of progesterone receptors in connective tissue cells of the lower nasal turbinates in women]. AB - BACKGROUND: The syndrome of the Rhinopathia gravidarum has been frequently discussed in the literature, but the etiology is yet unknown. An increase of oestrogen and progesterone concentration is said to contribute to the pathogenesis. The aim of this study was to localize estrogen (ER) and progesterone receptors (PgR) in the nasal mucosa of women and to compare the localization with the distribution of mast cells (MC). The patients' medical history was obtained with special emphasis on nasal symptoms during pregnancy, the menstrual cycle, or with the use of oral contraceptives. METHODS: Immunohistochemistry (IHC) was performed on formalin-fixed paraffin sections (nasal mucosa, inferior turbinate of 40 women) with monoclonal antibodies against ER, PgR, and mast cell tryptase. RESULTS: PgR-positive cells were found in fibroblasts (nuclear staining). The cytoplasmic staining for ER in serous glands and excretory ducts and for PgR in the interstitium of glands is considered nonspecific. The pattern of the receptor distribution was different from the pattern seen in the MC-IHC. No significant statistical results were obtained comparing the patient's medical histories and the immunohistochemical findings. CONCLUSIONS: Our findings possibly indicate a direct influence of progesterone on fibroblasts and therefore on the consistency of the extracellular matrix. Additionally, estrogen and progesterone might cause rhinopathic symptoms indirectly by changing the concentration of neurotransmitters (e.g. substance P, NO) and their receptors. PMID- 11383119 TI - [Cytotoxicity and genotoxicity of fluorides in human mucosa and lymphocytes]. AB - BACKGROUND: Fluorides are widely used in dental health products and drinking water, due to their beneficial effects in caries-prophylaxis and -treatment. Nevertheless, irritation of the gingiva and oropharyngeal mucosa as well as in gastric mucosa is observed since neither local nor systemic application is restricted to the teeth. These effects may partly be attributed to a known cytotoxicity of fluorides. Whether fluorides also have genotoxic effects on human mucosa or lymphocytes as a possible factor in tumor initiation was investigated in this study. MATERIAL AND METHODS: Human oropharyngeal epithelial cells and peripheral lymphocytes were incubated after single cell preparation with the aminefluoride Olaflur at concentrations of 2 ppm, 21 ppm, 35 ppm, 71 ppm and 213 ppm. The extent of cytotoxicity was investigated using the trypan blue exclusion test. Following incubation, electrophoresis for migration of DNA fragments, fluorescence staining and digital image analysis according to a standard protocol of the single cell microgel electrophoresis assay (Comet assay) followed. DNA damage was characterized using the Olive Tail Moment (OTM). RESULTS: For fluoride concentrations of 2 ppm to 35 ppm, non vital cells of less than 10% could be shown. After incubation with 71 ppm and 213 ppm Olaflur, there were 15% and 43% of damaged cells, respectively. Weak genotoxic effects on mucosal cells as well as on lymphocytes could be demonstrated at all concentrations tested. In fluoride concentrations of 213 ppm genotoxicity increased to max. OTM-levels of 23. CONCLUSIONS: Beside the cytotoxic effect of fluorides, also a minor genotoxic impact on human mucosa and on peripheral lymphocytes could be demonstrated using the Comet assay. Further investigations are warranted to examine fluorides in a model allowing for repeated or long term incubations on structurally intact human mucosa in vitro. Such a model will help to distinguish between DNA damage that may be repaired successfully and other impairments that may show an additive character in repetitive or chronic exposure in vivo. PMID- 11383120 TI - [IL-2 gene therapy in ENT carcinomas]. AB - Suppressed cellular immunity is common in patients with squamous cell carcinoma of the head and neck (HNSCC). It was demonstrated in previous studies that administration of interleukin 2 (IL-2) results in enhanced antitumoral immunity in vitro as well as in vivo. Since the serum half-life of IL-2 is relatively short, repeated applications are necessary to achieve therapeutically effective serum concentrations, but this strategy might cause severe side effects. Therefore, methods that provide high local cytokine levels over a prolonged period of time without the need for repeated injections are desirable. Gene therapy as an innovative treatment approach using tumor cells stably transduced to produce IL-2 might meet these criteria. In vitro manipulated tumor cells, if readministered in the vicinity of non-manipulated tumor cells, may enhance a specific anti-tumor response in vivo without systemic side effects. The present manuscript reviews the current literature dealing with IL-2-protein and -gene therapy with special emphasis on head and neck cancer. Our own in vitro results with IL-2 gene therapy in conjunction with published data from other authors argue in favour of an in vivo approach for this therapeutic strategy that is currently in progress in our department. PMID- 11383121 TI - [Carcinoma of the external ear canal and middle ear as interdisciplinary challenge for ear surgery and radiotherapy]. AB - BACKGROUND: Carcinoma of the external auditory canal are tumours considered to have a poor prognosis. Improvement of the survival rate by surgical means alone is not possible. Individual therapy modalities as a result of an interdisciplinary approach between otosurgeon and radiotherapist are necessary. PATIENTS AND METHODS: A series of 30 patients (3 patients pretreated at other institutions) with carcinoma of the external auditory canal and middle ear treated between 1978 and 1997 in our institutions was analysed with particular reference to tumour size and its relation to surrounding tissues, patterns of neck node involvement, surgical procedures, and radiation techniques. Clinical endpoints were freedom from local failure, overall survival, disease-free survival. The mean follow-up was 4.7 years (range: 0.1 to 18.8 years), median 3 years. RESULTS: Treatment by surgery and radiotherapy resulted in an overall 5 year survival rate of 51%. According to Pittsburgh classification the 5-year survival rate for early disease (T1- and T2-tumours) was 89%, for stage III 67% and for stage IV 39%. Most important prognostic factors were dural infiltration (all patients with dural invasion died within 2.2 years) and the infiltration of surgical margins (the 5-year survival rate of patients with complete tumour resection was 100%, but 54% in patients with tumour beyond surgical margins). 192 iridium HDR afterloading brachytherapy based on 3D CT-treatment planning is an effective tool in the management of local recurrences following surgery and a full course of external beam radiotherapy. CONCLUSIONS: Surgical resection followed by radiotherapy adapted to the stage of disease and grade of resection is the preferred treatment of cancer of the external auditory canal and middle ear. PMID- 11383122 TI - [Improved tumor site and contrast resolution in ultrasound diagnosis by using harmonic frequencies]. AB - BACKGROUND: The aim of new techniques in head and neck sonography is to increase the sensitivity and the specificity of the examination. With tissue harmonic imaging (THI) and contrast harmonic imaging (CHI) new techniques are available which allow increased contrast and resolution in head and neck sonography as well as a better detection of small blood vessels. METHODS: We studied whether these techniques improve sonographic detection of head and neck lymph nodes and primary tumours of the upper aerodigestive tract. RESULTS: The results indicate that THI allows a better detection of cervical lymph nodes. Furthermore, intranodal structures and the borders of the lymph node can be better detected. Using CHI, typical patterns of vascularization can be seen, allowing a better interpretation of the node's tumour status. Primary tumours can be detected sufficiently well using conventional B-scan techniques although they are easier to detect with THI. Using CHI, many tumour vessels can be detected, but this does not increase the sensitivity of the sonographic examination. CONCLUSION: THI and CHI may increase the specificity of sonographic diagnostics of cervical lymph nodes but show no clinical benefit regarding the diagnostics of primary tumours of the upper aerodigestive tract. PMID- 11383123 TI - [Experiences with using the ultrasound contrast medium levovist in differentiation of cervical lymphomas with color-coded duplex ultrasound]. AB - AIM: Proof of intranodal vessels in lymph nodes of the head and neck region and their association to specific types of vascularisation enables categorization of lymph nodes and improves preoperative predictability of tissue dignity. In many lymph nodes imaged by colour duplex sonography no intranodal vascularisation can be found. The purpose of the study was to find out if sonographic categorisation of lymph nodes, which do not show intranodal vessels in normal colour duplex sonography, is enabled with the contrast amplifier Levovist. METHOD: In 30 patients with metastasis in lymph nodes, malignant lymphomas or inflammatory lymph nodes with no or poor intranodal vascularisation, we administered Levovist, a contrast amplifier given intravenously, during colour duplex sonography. RESULTS: Intranodal vascularisation could be visualized in all cases by the use of Levovist contrast amplifier during the sonographic investigation. CONCLUSION: Use of the contrast amplifier Levovist leads to an improvement of the categorisation of lymph nodes. Especially inflammatory lymph nodes show more intranodal vessels in the hilus area, whereas metastatic lymph node vessels show no link to the hilus area. Preoperative diagnostic procedures may take advantage of this new technique. PMID- 11383124 TI - [Toxicity of glass ionomer cement]. AB - BACKGROUND: The hybrid bone substitute ionomeric cement achieves a stable and durable space-free bond to adjacent bone during hardening. Clinical studies have evaluated the material differently: Fully hardened ionomeric cement showed in middle ear surgery, e.g. as an ossicular prosthesis, good biocompatibility with outstanding functional results. In a few cases, freshly mixed ionomeric cement led to severe complications after contact with CSF in skull base surgery. Therefore we intended to evaluate the influence of early fluid contact on the quality of cement and to define the interval for a safe application of the material, using a cell culture model. Further we intended to investigate whether combining cement with homologous and alloplastic materials influenced its quality. METHODS: 1) Ionomeric cement (Ionocem) test bodies were placed in Ringer's solution at different times after the mixing phase. 2) Ionomeric cement (Ionocem) test bodies were coated with different clinically used homologous and alloplastic materials during the setting and hardening phase. The concentrations of released cement-forming ions and the toxic effects on mouse fibroblasts within cell cultures were measured. RESULTS: Cytotoxic effects were observed when ionomeric cement was not carefully protected from fluid contact for the first two hours after mixing. This was due to forced fast elution of large amounts of cement-constituting fluoride ions and aluminium ions and to the development of acid valences and their interactions. A total hardening time of less than 25 min had an especially unfavourable influence on cell behaviour. Cell impairments could be reduced significantly by coating the 30-minute cured cement with PDS sheeting and significantly by covering it with viscous collagene. On the other hand, cement toxicity was intensified in part by combinations with some other coating materials. CONCLUSIONS: Ionomeric cement should be kept dry and protected from any fluid contact for at least 30 minutes after mixing. Contact with soft tissue should also be avoided for this time. With a hardening time of 30 minutes, the quality and biocompatibility of glass ionomeric cement could be substantially optimized by coating it with PDS sheeting. These results should be verified in animal experiments and clinical trials. PMID- 11383125 TI - [Interesting case no. 44. Osteoma]. AB - We report on a 30-year-old man who was treated in the outpatient clinic for bifrontal headache for four weeks. Computed tomography showed sinusitis maxillaris, a large osteoma in the ethmoid sinus on the right side and a pneumocephalus. The surgical procedure included removal of the focus (sinusitis), extirpation of the osteoma and reconstruction of the skull base from an external approach. PMID- 11383126 TI - [Phoniatric aspects in microsurgical removal of benign vocal cord changes]. AB - INTRODUCTION: Microsurgery of benign vocal cord lesions is usually done to improve, to restore or to preserve patients' voice. Solid anatomical and physiological knowledge concerning the voice are basic requirements of any surgeon operating in this field. PREOPERATIVE DIAGNOSTICS AND DOCUMENTATION: The significance of patients' history as well as phoniatric investigation, laryngoscopic and stroboscopic findings and their proper documentation are presented. INDICATIONS: The surgical procedure must be part of an extensive therapeutical concept that also takes into consideration drug therapy as well as voice therapy procedures to improve voice quality. POSTOPERATIVE REGIMEN: The care for the patient's voice usually goes beyond surgery. To gain the best possible postoperative results, several steps are necessary in addition to surgical procedures. These steps will be pointed out. PMID- 11383127 TI - [Surgery of the mediastinum. III--Mediastinoscopy]. PMID- 11383128 TI - Help from my friends. The high court's marijuana ruling won't play in Mendocino. PMID- 11383129 TI - An equine mystery in the bluegrass. An epidemic is killing Kentucky's finest foals. PMID- 11383130 TI - The seat of identity. Bizarre brain changes. PMID- 11383131 TI - Your cholesterol might have to come down--way down. PMID- 11383132 TI - To fight off Alzheimer's, train your brain and take care of your body. PMID- 11383133 TI - Effect of cisplatin treatment on the urinary excretion of guanidinoacetic acid, creatinine and creatine in patients with urinary tract neoplasm, and on superoxide generation in human neutrophils. AB - Production of guanidinoacetic acid, a precursor of creatinine is known to be reduced by metabolic disturbance when kidney function is damaged, and thus it may be a sensitive marker of renal damage. Therefore, the urinary levels of guanidinoacetic acid, creatinine and creatine from patients with urinary tract neoplasm who received cisplatin treatment were measured by liquid chromatography mass spectrometry. Following the administration of cisplatin, the urinary excretion of guanidinoacetic acid decreased significantly, and the low concentration was maintained for at least five days. The concentrations of creatinine and creatine gradually decreased until the third day after cisplatin administration, and slightly increased on the fifth day. As superoxide might be concerned in renal damage by cisplatin, the effect of cisplatin on superoxide generation was also investigated using human neutrophils. Cisplatin significantly enhanced phorbol 12-myristate 13-acetate-induced superoxide generation in a concentration-dependent manner, but had no effect on the superoxide generation induced by N-formyl-methionyl-leucyl-phenylalanine and arachidonic acid. The superoxide generation increased by cisplatin was inhibited by staurosporine, an inhibitor of protein kinase C, but was rather enhanced by genistein, an inhibitor of protein tyrosine kinase. PMID- 11383134 TI - Protective effect of L-arginine against lipid peroxidation in goat epididymal spermatozoa. AB - L-arginine plays an important role in physiology of spermatozoa and is shown to enhance the metabolism of these cells. We report here the effect of L-arginine on membrane lipid peroxidation of goat epididymal spermatozoa. Both natural peroxidation as well as that induced by UV radiation, freezing and oxidizing agents have been studied. Irrespective of the nature of induction of peroxidation, L-arginine reduces the extent of lipid peroxidation in a concentration dependent manner. Both L-arginine and alpha-tocopherol act synergistically in protecting against lipid peroxidation induced by the above methods. Thus, in order to provide protection against lipid peroxidation, L arginine may be added in media used to preserve spermatozoa. PMID- 11383135 TI - Cystamine transport in spheroplasts of Saccharomyces cerevisiae. AB - This work is the first demonstration that cystamine is actively accumulated in spheroplasts of Saccharomyces cerevisiae. We have identified and quantitatively determined the transported cystamine in extracts of spheroplasts that have been incubated over different time periods and in the presence of different amounts of cystamine. The method used, already reported in literature for the identification of natural aliphatic polyamines in biological fluids, consists of a derivatization of spheroplast extracts with dabsyl-chloride and subsequent chromatographic analysis in HPLC. Our results show that cystamine accumulation is a function of time, it increases up to 2.5 min then decreases. Transport is inhibited by natural aliphatic polyamines, which, at the same concentration of cystamine (1 mM), cause a decrease in cystamine transport of about 90% for spermidine, 50% for spermine and only 15% for putrescine. Furthermore, transport is energy-dependent as demonstrated by a significant decrease observed in the presence of 2,4-dinitrophenol, ouabain and vanadate. In particular 0.2 mM ouabain causes a decrease of more than 60% in cystamine transport. Our data suggest that cystamine is transported in Saccharomyces cerevisiae spheroplasts via the same polyamine transport system(s) known to be operating in higher eukaryotic cells. PMID- 11383136 TI - Sensitized photooxygenation and peroxidase-catalyzed inactivation of xanthine oxidase--evidence of cysteine damage by singlet oxygen. AB - Xanthine oxidase (XO) has been investigated for its decreased activity in several cancerous tissues and constitutive generation of reactive oxygen species (ROS) in vivo seems to contribute significantly to its inactivation. Singlet oxygen (1O2) production has been suggested to be relevant when considering folic acid metabolism by cancer cells. Thus, the susceptibility of XO to inactivation by 1O2 generated either by the bioenergized systems folic acid/peroxidase/GSH/Mn2+/O2 and malonaldehyde/peroxidase/Mn2+/O2 or by methylene blue (MB) or eosin sensitized photooxygenation was studied. Our results showed that other ROS were also responsible for XO inactivation when MB was used. In contrast, eosin produced almost exclusively 1O2. Kinetic studies of XO oxidation in the malonaldehyde/peroxidase system showed that histidine (His) is a competitive inhibitor with respect to XO. A similar result was observed in the eosin photosensitized process, suggesting the involvement of 1O2 in both processes. In addition, an efficient quenching of XO oxidation by guanosine in the folic acid/peroxidase system was observed. Amino acid analysis revealed that cysteine (Cys) is more affected than other XO amino acids also prone to oxidation such as tyrosine (Tyr), methionine (Met) and His. These results indicate that 1O2 may cause oxidative damage to the Cys residues of XO, with loss of enzyme activity. Alteration of the flavin prosthetic site is hypothesized. PMID- 11383137 TI - Dielectric properties of some components of fruit aroma in carbon tetrachloride solution at 298.15 K. AB - The static electric permittivity and refractive index of carbon tetrachloride solutions of ethyl-2-methyl butanoate, 3-hexene-l-ol, 2-hexenal, ethyldecanoate, methyl anthranilate, anethole, damascenone and isoamyl propanoate were measured at 298.15 K in the concentration range up to 4.0 mol dm-3. The molar polarizations and refractions of the solutions, the partial molar polarizations and refractions of the solutes, as well as the dipole moments at infinite dilution of the solutes were calculated. On the basis of the concentration dependence of apparent values of the square of the molecular dipole moment, it was established that for the systems studied oligomeric species with low dipole moment were prevalent. With the assumption that the dipole moment of dimeric species amounts to zero (except for 3-hexene-l-ol and methyl anthranilate), the constant of dimerization was calculated and a weak molecular association was deduced. PMID- 11383138 TI - Evidence of a new type of protein-protein interaction: desensitized actomyosin blocks Ca(2+)-sensitivity of the natural one. A possible model for an intracellular signalling system related to actin filaments. AB - Actin filaments are certainly believed to function as an intracellular signalling system; however, this is not confirmed by direct evidence. We used a two-layer actomyosin gel with a concentration gradient of the troponin-tropomyosin complex (TT-complex, Ca(2+)-sensitive system) between the two layers. To prepare one layer of the system, natural actomyosin (nAM) rich in TT-complex was used. To prepare the second layer, we used desensitized actomyosin (dAM) without the complex. All experimental studies were made in medium with a low ionic strength. Two phenomena were observed: (1) dAM blocks Ca(2+)-sensitivity of nAM when the dAM weight portion in the system (as well as in mixed nAM + dAM suspension) reaches 40% and more; further increase of the dAM portion does not affect the Ca(2+)-sensitivity; (2) it was electrophoretically shown that a rapid diffusion of the TT-complex from nAM gel into the dAM gel took place. The apparent diffusion coefficient for the TT-complex in dAM gel is about (1-4).10(-4) cm2/sec, i.e. three orders higher than the same values for protein diffusion in water. PMID- 11383139 TI - Physics in muscle research. AB - Muscle is one of few organs whose performance can be measured by physical quantities. However, very few attempts have been made to apply theoretical physics to muscle. In this paper we will see how physical principles can be applied by taking advantage of unique properties of muscle structure. The first topic is to establish the stability conditions of sarcomere structure. The conclusions are then compared to some experimental facts. Next, we move on to the field theory fundamentals. The concept of energy density as a stress tensor is shown to be a powerful tool for the dielectric force theory to understand how proteins move under electric fields. By combining the structural stability theory and the dielectric force theory we arrive at a helical dipole array. We discuss the source of strong dipole fields and how the dipole strength could be controlled by Ca ions. The behavior of water and ions under electric fields is briefly discussed. The third topic is the mechanical stiffness of muscle in longitudinal and lateral directions. Some experimental data are shown and the physics of anisotropic stiffness is discussed. An appendix is provided to explain the pitfalls of experimenting with isolated components rather than organized structures (sarcomere). PMID- 11383140 TI - [Reasons for the necessity of health care reform]. PMID- 11383141 TI - [Specialization of the hospital psychiatric department]. PMID- 11383142 TI - [Role of the hospital psychiatric department in health care service-- results of a questionaire]. PMID- 11383143 TI - [Therapy and care of patients with substance-induced mental disorders- experiences in the hospital psychiatric departments]. PMID- 11383145 TI - [Care of patients with substance-induced mental disorders within a community mental health service center]. PMID- 11383144 TI - [Treatment programs for patients with substance-induced mental disorders]. PMID- 11383146 TI - [Care of patients with substance-induced mental disorders]. PMID- 11383147 TI - [Japanese Society of Psychiatric History]. PMID- 11383148 TI - [Pathology of intractable epilepsy with regard to surgical treatment]. AB - There have recently been a number of new pathological findings of specimens from epileptic foci that have increasingly been highlighted as opportunities for surgical treatment. These findings include a wide range from hippocampal sclerosis-related lesions of temporal lobe epilepsy to newly established malformative lesions. As to the former, special interest has been paid to additional knowledge regarding the pathology inside and outside the medial temporal regions, such as the significance of 1) accumulation of corpora amylacea in hippocampal sclerotic lesions, 2) dispersion of the granular layer of the dentate gyrus, 3) various pathologic features in the lateral temporal lobe, and 4) dual pathology in extratemporal areas. As increasing opportunities for surgical treatment of intractable epilepsy, new categories of malformative lesions are being established, including focal cortical dysplasia and microdysgenesis, the latter of which can be diagnosed only by pathological examination after surgery. In the field of surgical pathology of epilepsy patients, further clues to clarify the pathomechanism of epilepsy remain to be discovered. PMID- 11383149 TI - [Problems surrounding offenders with mental disorder--concerning the symposium "the present subjects of forensic psychiatry" of the 95th Congress of the Japanese Society of Psychiatry and Neurology]. PMID- 11383150 TI - [Automated analysis for determination of PCBs in fish]. AB - An automated analytical method for determination of polychlorinated biphenyls (PCBs) in fish was developed using supercritical fluid extraction (SFE) in combination with an automated sample preparation instrument (Prep) and GC/MS. By incorporating basic alumina with the sample in the extraction process, and optimizing the amount of carbon dioxide used, fish lipid was selectively reduced. The extract was cleaned up on a Florisil cartridge with Prep. The method was evaluated using naturally contaminated tissues and by comparison of automated analytical method results with those obtained by the conventional method. Mean recovery of PCBs from 3 kinds of fish including hairtail, mackerel and yellowtail were 69.8%, 90.2% and 81.1%, respectively. This method is less laborious and requires far less organic solvent than the conventional method, but produced comparable results. PMID- 11383151 TI - [Cause of bisphenol A migration from cans for drinks and assessment of improved cans]. AB - In the previous investigation, we found that some cans for coffee and black tea drinks released large amounts of bisphenol A (BPA) into their contents. Equivalent cans were obtained and the cause of BPA migration was investigated. Equivalent cans A, B and D contained high levels of BPA in the side seam, in the bottom, and in the bottom and the side seam, respectively, while can C contained some level of BPA in the body, which has a large area, therefore, all of them contained high amounts of BPA in their coatings. In the migration test, there was no BPA migration from the cans into water at 60 and 95 degrees C for 30 min, into 20% ethanol at 60 degrees C for 30 min, or into n-heptane at 25 degrees C for 60 min. However, at 120 degrees C for 30 min, equivalent cans released 35-124 ng/mL BPA into the water. The total migration was similar to the total residues of BPA in the can coating and was close to the total amount of BPA in the drinks. Thus, BPA migration from the can coating requires heating to more than 105 degrees C, which is the glass transition temperature of the epoxy resin. Improved cans which contained less than 1/10 as much BPA as the equivalent cans showed very low migration levels, i.e., 3-6 ng/mL. PMID- 11383152 TI - [Estimation of daily dietary intake of aluminum]. AB - The daily dietary intake of aluminum was estimated through a total diet study from 1996 to 1998. In ten institutes, total diet study samples were prepared and their aluminum concentration was determined. The average daily intake of aluminum was 3.5 mg and the range was 1.8-8.4 mg. The validity of the analytical result was supported by analyses of certified reference materials. PMID- 11383153 TI - A multiplex PCR method of detecting recombinant DNAs from five lines of genetically modified maize. AB - Seven lines of genetically modified (GM) maize have been authorized in Japan as foods and feeds imported from the USA. We improved a multiplex PCR method described in the previous report in order to distinguish the five lines of GM maize. Genomic DNA was extracted from GM maize with a silica spin column kit, which could reduce experimental time and improve safety in the laboratory and potentially in the environment. We sequenced recombinant DNA (r-DNA) introduced into GM maize, and re-designed new primer pairs to increase the specificity of PCR to distinguish five lines of GM maize by multiplex PCR. A primer pair for the maize intrinsic zein gene (Ze1) was also designed to confirm the presence of amplifiable maize DNA. The lengths of PCR products using these six primer pairs were different. The Ze1 and the r-DNAs from the five lines of GM maize were qualitatively detected in one tube. The specific PCR bands were distinguishable from each other on the basis of the expected length. The r-DNA could be detected from maize samples containing 0.5% of each of the five lines of GM maize. The sensitivity would be acceptable to secure the verification of non-GMO materials and to monitor the reliability of the labeling system. PMID- 11383154 TI - Comparison of TaqMan Salmonella amplification/detection kit with standard culture procedure for detection of Salmonella in meat samples. AB - We evaluated the TaqMan PCR Salmonella amplification/detection kit (PE Applied Biosystems) for rapid detection of Salmonella from a variety of meat samples. This system uses the 5' nuclease activity of Taq DNA polymerase, which digests an internal fluorogenic probe to monitor the amplification of the target gene. The detection sensitivity of the kit, using 2 kinds of DNA extraction protocols, was compared with that obtained with 4 protocols of official culture methods. A total of 98 meat samples (16 raw beef, 31 pork and 51 chicken) were tested. The results of the TaqMan PCR method and the combined results of the 4 cultural protocols showed excellent agreement. However, no single culture protocol showed optimal recovery of Salmonella comparable to the PCR method. These results suggest that the TaqMan PCR method is a reliable and rapid method useful for detecting Salmonella in meat products. PMID- 11383155 TI - [Determination of neohesperidin dihydrochalcone in foods]. AB - An HPLC method was developed for the determination of neohesperidin dihydrochalcone (NHDC) in foods. A solid sample was extracted with methanol. The extract was evaporated and the residue was dissolved in methanol-water (2:8) mixture. In the case of fluid sample, the sample solution was prepared by dilution with methanol-water (2:8) mixture. The sample solution was cleaned up on a C18 cartridge. The cartridge was washed with water and methanol-water (3:7) mixture, and NHDC was eluted with methanol-water (7:3) mixture. NHDC in the eluate was separated on an ODS column and determined with a UV detector (282 nm). The identity of NHDC in foods was confirmed by means of HPLC with a photodiode array detector. The recoveries of NHDC added to various kinds of foods were 75.2 134%. The determination limit of NHDC was 1 microgram/g. PMID- 11383156 TI - [Determination method of isocitric acid in food additive citric acid]. AB - A simple and rapid method using HPLC was developed for the determination of isocitric acid in food additive citric acid. One gram of sample was dissolved in 100 mL of water. HPLC separation was performed on an Inertsil ODS-3 column (4.6 mm i.d. x 250 mm) using 0.1% phosphoric acid as the mobile phase at a flow rate of 1 mL/min. Isocitric acid was detected at 210 nm. The calibration graph was rectilinear from 5 to 100 micrograms/mL. The recoveries of isocitric acid from sample at the levels of 0.1% and 0.4% were 98% and 99%, respectively, and the determination limit was 0.05%. PMID- 11383157 TI - [Contents of phthalate in polyvinyl chloride toys]. AB - The presence of plasticizers in PVC toys obtained in October 1998 was investigated. Diisononyl phthalate (DINP), di-2-ethylhexyl phthalate (DEHP), di-n butyl phthalate (DBP), dinonyl phthalate (DNP), diheptyl phthalate (DHP), and di 2-ethylhexyl adipate (DEHA) were detected. The phthalates were found in all of the 68 samples. The principal phthalate found in toys was DINP, which was present in 48 of 68 samples. The DINP content ranged from 15 mg/g to 580 mg/g, and mean content was 308 mg/g. The highest content was found in a pacifier toy. DEHP was present in 20 of 68 samples and the content ranged from 2.0 mg/g to 380 mg/g. The mean content was 162 mg/g. It was found in 60% of domestic toys. PMID- 11383158 TI - [Application and improvement of aflatoxin analysis in foods using a multifunctional column and HPLC]. AB - In an earlier report, we developed a rapid, sensitive and clean method consisting of non-chloroform extraction, clean-up on a commercial multifunctional cartridge column and HPLC with fluorescence detection for the analyses of aflatoxins. In this report, we applied this method to analyze aflatoxins in nuts, giant corn, cereals, spice and black teas. The method was effective for macadamia nuts, walnuts, hazelnuts, brazil nuts, giant corn, rice, wheat and buckwheat, and the recoveries of aflatoxins B1, B2, G1 and G2 spiked in them at the level of 10 ng/g were 85-106%. However, in the chromatograms of spices and black tea, many background peaks were observed. Therefore, we added a purification step with an affinity column to the clean-up of these samples with the multifunctional cartridge column. After the additional purification, most of the background peaks were gone. The recoveries of aflatoxins B1, B2 and G1 spiked at the level of 10 ng/g were 71-112% except for the case of B2 in white pepper (48%). The recoveries of G2 were 49-95%. PMID- 11383159 TI - Flavor production from a non-stick oil by moulds. AB - Natural flavor was accidentally produced from rice cake products in Japan. A non stick oil had been sprayed on the products during the production process. It was found that a Penicillium corylophilum strain, a contaminant of the oil, produced the flavor from the oil. The ingredients of the flavor were four volatile substances, 2-heptanone, 2-nonanone, 2-heptanol, and 2-nonanol. Challenge tests with the mould strain in a rice cake system were performed under various conditions. The volatile substances were produced in the largest amounts at 25 degrees C, followed by 20 or 30 degrees C then 10 degrees C. 2-Heptanone was produced most remarkably at 25 degrees C, followed by 2-nonanone, 2-heptanol, and 2-nonanol. The growth patterns of the mould were similar between 20-30 degrees C, and the growth at 10 degrees C was delayed. The non-stick oil itself had neither flavor nor volatile substance. The flavor was also produced from coconut oil, which was one of the materials of the non-stick oil. No bacteria or yeasts tested produced any flavor from the non-stick oil, whereas most of the moulds tested produced flavor components. PMID- 11383160 TI - [Application of immunoaffinity columns to food analysis]. PMID- 11383161 TI - [Geographic references of chemicals]. PMID- 11383162 TI - Ole tyme religion: Bravo is Bay Ridge; Bay Ridge is Bravo. PMID- 11383163 TI - Tornado warning. AB - Your ability to mitigate, prepare, respond and recover from a tornado touchdown greatly depends upon preplanning and the application of common-sense principles. You must be able to provide emergency medical assistance for your community when it needs you most. Develop an Emergency Tornado Operations Plan now. You'll be glad you did when the warning sirens wail. PMID- 11383164 TI - Fatal phrases. PMID- 11383165 TI - The state of EMS. PMID- 11383166 TI - Domestic preparedness: the grand illusion. AB - The problems posed by terrorism to not only the emergency response community, but to national security at large can be overwhelming. Adoption of what would be considered prudent and effective business practices by implementing a disciplined and effectively structured central strategy cannot be overencouraged. The emerging strategy must take into account the existing emergency response infrastructures and build upon existing capacity in an effort to achieve greater readiness. This technique is no different than the training and issuance of radiological response equipment to emergency responders in the 1950s by the then Civil Defense Agency. The training that is offered, especially to EMS providers, needs to be institutionalized to ensure that our peers, on a regular basis, revisit curriculum content. Incorporating a training module within the existing DOT NHTSA initial and refresher EMT and paramedic educational curricula could easily achieve this goal. Implementing fiscal support to the local emergency response agencies in a sustainable manner is a must. The costs associated with training, equipping and servicing the equipment and medication stores are budget busters. This is a threat to national security and, as such, the federal government needs to rise to the challenge of supporting the local response organizations that will meet this threat head-on during the aftermath of an attack. As previously mentioned, when the U.S. faced its last large national security threat (Soviet nuclear missiles), we witnessed the materialization of a comprehensive agenda that provided most of the attributes we desire with the contemporary problem of terrorism. There is no single solution to the problem of terrorism. In fact, it will take many individuals and functional areas to come together and stop viewing the threat as a "cash cow." The improved response capacity for acts of terrorism will have an inevitable "spillover benefit" of better trained and equipped emergency responders for everyday emergencies; which will inevitably be our "payday." PMID- 11383167 TI - The EMS safety officer: incident accountability. PMID- 11383168 TI - Documenting patient refusals. AB - Patient morbidity and mortality, subsequent to either patient- or provider initiated refusals, are noteworthy. It has been estimated that hospital admission is twice as likely after prehospital providers refuse a patient transportation to the hospital. In one particular study group, prehospital-provider refusal of transportation, as opposed to patient refusal of transportation, accounted for 73% of the post-refusal hospital admissions. Provider-initiated refusals are tantamount to a time bomb. There are very few justifiable provider-initiated refusals of treatment or transportation. If the call is of a non-emergency nature, the decision not to treat or transport should be a mutual agreement between the patient and provider. Consult with the online medical director for guidance as needed. Document the physician's name, consulting facility and medical direction. As with every patient encounter, a legally defensible runsheet should be completed. Should you write a report if your services are "not needed," or if you are canceled en route to the call? For your protection, a report archiving every run should be documented. If your services are canceled en route, note the canceling authority and time of cancellation. If your services are canceled at the scene, document the canceling authority, time of cancellation and the circumstance. It is important to specifically document that "no patient contacts were made." When patient contact is made, a patient-provider relationship is established, thereby redefining your duty to the patient. Protect yourself, your crew members, your chain of command, your jurisdiction and your agency by writing a legally defensible informed refusal report. According to one source, "Every negligence case in the last 30 years has been decided on its documentation." If it wasn't written down, it wasn't done. Be safe, and document safely. PMID- 11383169 TI - Crew resource management for EMS personnel. PMID- 11383170 TI - Noncardiac chest pain. PMID- 11383171 TI - Emergency baptism of the critically ill or stillborn child. PMID- 11383172 TI - The scene safe myth. PMID- 11383173 TI - The French connection: EMS in France. PMID- 11383174 TI - Kaiser does quality assurance and risk management in online discussion groups. PMID- 11383176 TI - Building the case for e-case management. PMID- 11383175 TI - Harvard's HealthAware project guides patients through healthcare decisions. PMID- 11383177 TI - Redefining interdisciplinary practice. PMID- 11383178 TI - Issues surrounding adjuvant chemotherapy for breast cancer during pregnancy. PMID- 11383179 TI - Into the millennium: open the door and let the future in for cancer nursing research. PMID- 11383180 TI - The payment of research subjects: ethical concerns. AB - PURPOSE/OBJECTIVES: To present models for the payment of research subjects, pros and cons of each, and the application of ethical principles in the current environment. DATA SOURCES: Federal regulations and guidelines, current and historical research, and opinions expressed in the nursing, medical, bioethics, pharmacology, and clinical trial literature. DATA SYNTHESIS: Relevant regulations are sparse. A variety of payment models exist, but all are somewhat problematic. Balancing the principles of respect for people and justice in our current society will not be entirely possible. Practice decisions must be made for individual situations. Special care must be taken with patients because they are subject to therapeutic misconception. CONCLUSIONS: No model fits all situations, but research nurses can use their skills to evaluate particular applications. As a society, nurses must promote participation in research as a socially responsible activity and prevent unethical payment models from predominating. IMPLICATIONS FOR NURSING PRACTICE: Oncology research nurses may be involved with the consent process, the development of protocols, and staff education. A thorough understanding of an issue that may impair the consent process or impinge upon subjects' rights is critical to effective and ethical practice. PMID- 11383181 TI - Educational needs and altered eating habits following a total laryngectomy. AB - PURPOSE/OBJECTIVES: To describe eating-related experiences and informational needs of people following total laryngectomies. DESIGN: Descriptive study. SETTING: Internet-based laryngectomy support group in the United States. SAMPLE: 34 people with a laryngectomy (68% total laryngectomy, 29% with total plus radical neck dissection, and 3% with partial laryngectomy with radical neck dissection): 29 males, 5 females; mean age of 62 years. METHODS: Members of a laryngectomy support group completed a Food Eating Experiences and Diet Questionnaire designed by the investigators. Both quantitative and qualitative data were collected. MAIN RESEARCH VARIABLES: Effect of laryngectomy on food choice, eating habits, and overall enjoyment of eating: perceptions of teaching received from healthcare professionals regarding potential eating difficulties as a result of laryngectomy. FINDINGS: 90% of the participants experienced a change in one or more aspects of eating. The most prominent changes were decreased sense of smell, decreased taste, decreased enjoyment of eating, and an increase in the length of time required to eat meals. Most participants were not satisfied with the information they received from healthcare professionals. Topics requiring emphasis during patient teaching were identified from participants' comments. CONCLUSIONS: Total laryngectomy produced significant changes in factors related to eating that can affect nutritional intake and quality of life. Participants reported that most healthcare providers did not adequately prepare them for potential alterations in eating that can occur following a total laryngectomy. IMPLICATIONS FOR NURSING PRACTICE: Data from this study can be used to raise awareness of incidence and severity of changes in eating that occur after total laryngectomy and to improve patient preparation to cope with these changes. PMID- 11383182 TI - Weight gain and quality of life in women treated with adjuvant chemotherapy for early-stage breast cancer. AB - PURPOSE/OBJECTIVES: To document weight gain in women treated with adjuvant chemotherapy for early-stage breast cancer and to examine the relationship of weight gain and perceived quality of life (QOL). DESIGN: Descriptive, correlational study. SETTING: Data collected in three settings: an ambulatory oncology service of a university teaching hospital, a private oncology office, and a university-affiliated health clinic, all located in southern New England. SAMPLE: Women with stage I or II breast cancer with primary treatment of simple or modified mastectomy or breast-conserving surgery with radiotherapy scheduled to receive adjuvant chemotherapy. METHODS: Weight data collected through retrospective chart review. QOL data collected prospectively using the Linear Analog Self Assessment Symptom Distress Scale for Breast Cancer and the Functional Assessment of Cancer Therapy for Breast Cancer Scale. MAIN RESEARCH VARIABLES: QOL and weight gain of five pounds or more. FINDINGS: One year after treatment began, 62.5% of the study participants experienced weight gain (X = 10.44 lb), with a range of 5-27 pounds. After two and three years, 68% and 40%, respectively, maintained a clinically significant weight gain. A greater weight gain occurred over time in premenopausal women. No correlation between overall QOL and weight gain existed, but selected items were significantly positively correlated with weight gain. CONCLUSIONS: This study documented a significant weight gain in women treated with adjuvant chemotherapy for early-stage breast cancer. A large percentage of those women maintained this weight gain. Women premenopausal at diagnosis had a greater tendency to gain weight. Although weight gain was not correlated with overall QOL it was distressing for these women. IMPLICATIONS FOR NURSING PRACTICE: Nurses can incorporate the possibility of weight gain into the plan of care for women with breast cancer. Nurses should include this information in education about side effects of treatment and in the ongoing nursing assessment of patients with breast cancer. PMID- 11383183 TI - Nurse documentation: not done or worse, done the wrong way--Part I. AB - PURPOSE/OBJECTIVES: To focus on nursing documentation and expanding technologies (e.g., facsimile, telephone, e-mail, computer charting) that offer different ways to record, deliver, and receive patient records and avoid nursing liability for inadequate or inaccurate documentation. DATA SOURCES: Nursing, non-nursing healthcare, legal journals, case law, and related Internet sources. DATA SYNTHESIS: To avoid liability for inadequate or inaccurate documentation, nurses must be aware of the major issues involved in documentation litigation. New technology is altering how healthcare documentation is done and raising new confidentiality issues. CONCLUSIONS: Nurses should follow their facility's guidelines and principles for documentation of patient care, especially when using more advanced technologies. IMPLICATIONS FOR NURSING PRACTICE: Educating nurses about the principles of documentation and the importance of implementing risk-reduction practices will help guard against liability and ultimately improve patient care. PMID- 11383184 TI - Backing and forthing: the process of decision making by women considering participation in a breast cancer prevention trial. AB - PURPOSE/OBJECTIVES: To describe the process of decision making by women considering participation in a breast cancer prevention trial (BCPT). DESIGN: Qualitative. SETTING/SAMPLE: Twenty-six women considering participation in a BCPT in the Northeastern United States. METHODS: Women were interviewed one or two times over a period of one year, with each interview averaging 40 minutes in length. The grounded theory method was used to collect and analyze the data. In depth interviews were conducted with each participant. Data were analyzed using the constant comparative method. MAIN RESEARCH CONCEPTS: Context, decision making, meaning. FINDINGS: The core variable of backing and forthing is a nonlinear complex process of decision making that includes reviewing life, wanting to be sure, chancing and deciding within the contexts of fear, view of self in the world, transgenerational issues, and social support. CONCLUSIONS: The process of decision making for women considering participation in a BCPT is complex. Women tend to make decisions based on what is in their heads and hearts. They often are concerned more about others than they are about themselves. IMPLICATIONS FOR NURSING PRACTICE: Trust in the provider and active involvement in the process is critical to women making a decision to participate in a BCPT. Decision making is unique for each woman; however, understanding the context, the core variables, and the process will help healthcare providers to support decision making. PMID- 11383185 TI - The relationship between access and stage at diagnosis of breast cancer in African American and Caucasian women. AB - PURPOSE/OBJECTIVES: To identify the relationships among potential access, realized access, and stage at diagnosis of breast cancer in African American and Caucasian women receiving care within an equal economic access healthcare system. DESIGN: Descriptive-comparative design. SETTING: Department of Defense Military Health System. SAMPLE: 62 African American and 573 Caucasian women (ages 25-97) diagnosed with breast cancer over a 10-year period. METHODS: Secondary analysis using tumor registry records data. MAIN RESEARCH VARIABLES: Economic, potential, and realized access and stage at diagnosis of breast cancer. FINDINGS: When compared to Caucasian women, African American women were diagnosed at later stages, were younger when diagnosed, were from a lower socioeconomic status, had a higher percentage of incidental breast self-examination-discovered abnormalities, and had a lower percentage of mammogram-discovered abnormalities. CONCLUSIONS: Economic access to care did not always result in early diagnosis. Other factors such as age, race, socioeconomic status, and means of discovery influenced diagnosis outcomes. IMPLICATIONS FOR NURSING PRACTICE: Interventions aimed at increasing participation in breast health programs also should focus on noneconomic aspects of access such as help-seeking behaviors and perceptions of access to care. Additional studies should be conducted to evaluate adherence to breast cancer screening guidelines by women receiving care within equal economic access healthcare systems. PMID- 11383186 TI - Ethnic influences on body awareness, trait anxiety, perceived risk, and breast and gynecologic cancer screening practices. AB - PURPOSE/OBJECTIVES: To examine ethnic influences on body awareness, trait anxiety, perceived risk, and breast and gynecologic cancer screening practices. DESIGN: Descriptive, correlational secondary analysis. SETTING: Urban and rural home and community populations. SAMPLE: 233 women: 138 (59%) Caucasian, 37 (17%) African American, 29 (12%) Hispanic, and 29 (12%) American Indian women (X = 46.86 years) were recruited through mailings, churches, and community organizations. METHODS: Structured questionnaires. MAIN RESEARCH VARIABLES: Body awareness, trait anxiety, perceived risk, and breast and gynecologic cancer screening practices. FINDINGS: Ethnicity predicted breast and gynecologic cancer screening practices (except clinical breast examination), body awareness, trait anxiety, and perceived risk. Hispanic and American Indian women reported greater breast self-examination frequency than Caucasian and African American women. Caucasian and African American women reported more mammogram use than Hispanic and American Indian women. Increased body awareness was related to fewer gynecologic exams for American Indian women. CONCLUSIONS: Women of different ethnic backgrounds respond differently to breast and gynecologic cancer screening practices. The influence of psychosocial variables on these practices varied with different groups. IMPLICATIONS FOR NURSING PRACTICE: Nursing interventions to increase breast and gynecologic cancer screening should be ethnic-specific, with particular attention to the meaning of body awareness to American Indian women and trait anxiety and perceived risk to African American women. PMID- 11383187 TI - The experience of women receiving brachytherapy for gynecologic cancer. AB - PURPOSE/OBJECTIVES: To explore and document the lived experience of receiving low dose rate brachytherapy for gynecologic cancer. DESIGN: Qualitative method based on phenomenology. SETTING: Radiation treatment facility in a cancer-care setting in Toronto, Ontario, Canada. SAMPLE: Ten women between the ages of 36 and 75 (x = 59.2) receiving low-dose rate brachytherapy for cancer of the cervix or endometrium. METHODS: Verbatim data were analyzed manually using Giorgi's method of analyzing qualitative data. FINDINGS: Three themes emerged from the data: (a) women's experiences with brachytherapy were embedded within the complete context in which treatment was given, shaped by personal, environmental, and treatment related factors, (b) the discomfort that women experienced during brachytherapy was perceived as a totality of symptoms including but not limited to pain, and (c) the brachytherapy experience was characterized by an intense focus on time and tensions embedded in issues related to time. CONCLUSIONS: When dealing with the brachytherapy treatment, women are concerned with the context in which the treatment is provided and the care that is associated with the treatment. Different and unique strategies assist women to get through treatment. Supportive nursing interventions can be implemented easily in the nursing care plan for women undergoing brachytherapy. IMPLICATIONS FOR NURSING PRACTICE: The aspects of nursing care that women perceive as positive, such as competence level of the nurse, symptom management, and providing information in sensory terms, should be strengthened. Alternatively, aspects of nursing care that are perceived negatively by women should be changed. Nurses have to avoid situations that will prolong the time of brachytherapy treatment. Nurses should support women in using coping strategies that assist them in getting through the brachytherapy treatment. PMID- 11383188 TI - Hearing and Perception. PMID- 11383189 TI - A prior for global convexity in local shape-from-shading. AB - To solve the ill-posed problem of shape-from-shading, the visual system often relies on prior assumptions such as illumination from above or viewpoint from above. Here we demonstrate that a third prior assumption is used--namely that the surface is globally convex. We use complex surface shapes that are realistically rendered with computer graphics, and we find that performance in a local-shape discrimination task is significantly higher when the shapes are globally convex than when they are globally concave. The results are surprising because the qualitative global shapes of the surfaces are perceptually unambiguous. The results generalise findings such as the hollow-potato illusion (Hill and Bruce 1994 Perception 23 1335-1337) which consider global shape perception only. PMID- 11383190 TI - The backward inclination of a surface defined by empirical corresponding points. AB - Three experiments were conducted to investigate whether a locus of binocular correspondence extends eccentrically from the vertical horopter. In experiment 1, we investigated whether the backward inclination of the vertical horopter was manifested in the angle at which readers prefer to orient the page. All observers preferred a page inclined backwards to any other orientation. This backward inclination was less than predicted from previous psychophysical reports, however. In experiment 2, we investigated the extent of binocular correspondence, defined by minimal apparent interocular horizontal motion, in the central 24 deg of the binocular field. Our data define a planar surface inclined top-away from the observer as a locus from which psychophysical corresponding points are stimulated. In experiment 3, we measured vertical adjustments required to eliminate apparent vertical motion. Together, the pattern of results from experiments 2 and 3 is most consistent with a planar surface, inclined top-away from the observer. This is consistent with Helmholtz's account of the backward inclination of the vertical horopter and expands the locus of zero horizontal disparity from a single line in the median plane to eccentric loci extending at least +/- 12 deg in the central binocular field. PMID- 11383191 TI - Ambiguity and the 'mental eye' in pictorial relief. AB - Photographs of scenes do not determine scenes in the sense that infinitely many different scenes could have given rise to any given photograph. In psychophysical experiments, observers have (at least partially) to resolve these ambiguities. The ambiguities also allow them to vary their response within the space of 'veridical' responses. Such variations may well be called 'the beholder's share' since they do not depend causally on the available depth cues. We determined the pictorial relief for four observers, four stimuli, and four different tasks. In all cases we addressed issues of reliability (scatter on repeated trials) and consistency (how well the data can be explained via a smooth surface, any surface). All data were converted to depth maps which allows us to compare the relief from the different operationalisations. As expected, pictorial relief can differ greatly either between observers (same stimulus, same task) or between operationalisations (same observer, same stimulus). However, when we factor out the essential ambiguity, these differences almost completely vanish and excellent agreement over tasks and observers pertains. Thus, observers often resolve the ambiguity in idiosyncratic ways, but mutually agree--even over tasks--in so far as their responses are causally dependent on the depth cues. A change of task often induces a change in 'mental perspective'. In such cases, the observers switch the 'beholder's share', which resolves the essential ambiguity through a change in viewpoint of their 'mental eye'. PMID- 11383192 TI - The influence of presentation format and viewer training in the visual arts on the perception of pictorial and aesthetic qualities of paintings. AB - The comparability of viewers' responses to slide-projected and computer-generated images of nine paintings by renowned artists to those obtained from individuals experiencing the originals in the galleries of the New York Metropolitan Museum of Art was investigated. The influence of training in the visual arts upon evaluative judgments made under the three presentation formats was also assessed. Specifically, art-trained and untrained participants in each format condition rated each artwork on sixteen measures of physical and structural characteristics, novelty of content, and aesthetic qualities. Analyses revealed significant differences in the judged hedonic value of the originals as contrasted with the two types of reproduction, whereas trained and untrained participants' evaluations of the pictorial qualities of the artworks were comparable across presentation formats. Findings are discussed in terms of a facsimile-accommodation hypothesis proposed by the authors. PMID- 11383193 TI - Classical and inverted White's effects. AB - In classical White's effect, intermediate-luminance targets appear lighter when they interrupt the dark stripes of a grating and darker when they interrupt the light stripes. The effect is reversed when targets are of double-increment or double-decrement luminance, relative to the luminances of grating stripes. To find a common explanation for classical and inverted effects, we ran two experiments. In experiment 1, we utilised intermediate-target displays to show that perceived transparency dominates over occlusion only when the target luminance is close to the luminances of top regions. This result weakens transparency-based accounts of White's effect. In experiment 2, we varied grating contrast and target luminance to measure the classical effect in seven intermediate-target cases, as well as the inverted effect in four double increment and four double-decrement cases. Both types of effect are explained by a common model, based on assimilation to the top region and contrast with the interrupted region, weighted by adjacency along the luminance continuum. PMID- 11383194 TI - Motion capture depends on signal strength. AB - When flickering dots are superimposed onto a drifting grating, the dots appear to move coherently with the grating. In this study we examine: (i) how the perceived direction of a compound stimulus composed of superimposed grating and dots, moving in opposite directions with equal speeds, is influenced by the relative strength of the motion signals; (ii) how the perceived speed of a compound stimulus composed of superimposed grating and dots, moving in the same direction but at different speeds, is influenced by the relative strength of the motion signals; and (iii) whether this stimulus is discriminable from its metameric speed match. Dot signal strength was manipulated by using different proportions of signal dots in noise and different dot lifetimes. Both the perceived direction and speed of these compound stimuli depended upon the relative motion-signal strengths of the grating and the dots. Those compound stimuli that appeared coherent were not discriminable from the speed-matched metameric compound stimuli. When the signals were completely integrated into a coherent compound stimulus, the local motion signals were no longer perceptually available, though both contributed to the global percept. These data strongly support a weighted combination model where the relative weights depend on signal strength, instead of a winner-takes-all model. PMID- 11383195 TI - Velocity not acceleration of self-motion mediates vestibular-visual interaction. AB - We investigated the influence of vestibular stimulation with different angular accelerations and velocities on the perception of visual motion direction. Constant accelerations resulting in different angular velocities and constant angular velocities obtained at different accelerations were combined in twenty healthy subjects. Random-dot kinematograms with coherently moving pixels and randomly moving pixels were used as visual stimuli during whole-body rotations. The smallest percentage of coherently moving pixels leading to a clear perception of motion direction was taken as the perception threshold. Perception thresholds significantly increased with increasing angular velocity. Increased acceleration, however, had no significant effect on the perception thresholds. We conclude that the achieved angular velocity, and not acceleration, is the predominant factor in the processing of vestibular-visual interaction. PMID- 11383196 TI - Similarity between Petter's effect and visual phantoms. AB - Here we draw attention to similarity between Petter's effect and the visual phantom illusion. Phantoms are visible when the spatial frequency of the inducing grating is low or the occluder is thin, whereas phantoms are invisible when the spatial frequency of the inducing grating is high or the occluder is thick. Moreover, phantoms are perceived in front of the occluder when they are visible, whereas the occluder is seen in front of the inducing gratings when phantoms are invisible. These characteristics correspond to Petter's effect, in which the thicker region tends to be perceived in front of the thinner region when two regions of the same lightness and of different sizes overlap, since 'thick' corresponds to low spatial frequency of the inducing grating or a thick occluder while 'thin' corresponds to high spatial frequency of the inducing grating or a thin occluder. PMID- 11383197 TI - The privacy paradox: the divergent paths of the United States Supreme Court and state courts on issues of sexuality. PMID- 11383198 TI - The pharmacist duty to warn revisited: the changing role of pharmacy in health care and the resultant impact on the obligation of a pharmacist to warn. PMID- 11383199 TI - Building a bridge to cross the chasm. PMID- 11383200 TI - Lessons learned: fires in the home care setting. PMID- 11383201 TI - Aligning environment of care requirements. PMID- 11383202 TI - Building a "good practices" database. PMID- 11383203 TI - Fine tuning the tailored survey policy. PMID- 11383204 TI - [Application of novel drug delivery system, fusogenic liposome, for cancer therapy]. AB - Over the past decade, many studies concerning novel anti-cancer therapies have been reported. It has been occasionally noted that a powerful anti-cancer drug, especially one whose target is the cytoplasm or cell nucleus, does not work due to the low permeability across a plasma membrane, degradation by lysosomal enzymes through an endocytosis-dependent pathway, and other reasons. Thus, several approaches using drug delivery systems (DDS) are focused on overcoming these difficulties, eventually leading to the induction of maximal ability of anti-cancer drug. In this respect, we have developed a new paradigm for cancer therapy using a novel drug delivery system, fusogenic liposome. Fusogenic liposomes are composed of the ultraviolet-inactivated Sendai virus and conventional liposomes. Fusogenic liposomes effectively and directly deliver their encapsulated contents into the cytoplasm using a fusion mechanism of the Sendai virus, whereas conventional liposomes are taken up by endocytosis. Thus, fusogenic liposome is a good candidate as a vehicle to deliver drugs into the cytoplasm in an endocytosis-independent manner. In this report, we show the feasibility of fusogenic liposome as a delivery vehicle for anti-cancer drugs using a fragment A of diphtheria toxin as an anti-cancer reagent. We also demonstrate the application of fusogenic liposome for cancer gene therapy and cancer vaccines using a TNF-alpha-expression plasmid and a chicken egg ovalbumin, respectively. PMID- 11383205 TI - [Tumor angiogenesis and tumor angiogenesis inhibitors]. AB - It is important to survey the molecular targets which are involved in tumor angiogenesis for the development of antiangiogenic agents as one of the cancer therapy. This article is meant to review the recent molecular targets of tumor angiogenesis and the molecular mechanism of antiangiogenic agents in human clinical trials. PMID- 11383206 TI - [Signal transduction inhibitor]. AB - Numerous molecular targets for cancer chemotherapy have been identified based on the progress made in molecular biology, and new categories of anticancer drugs have been developed. They are variously called target-based drugs, non-cytotoxic drugs and cytostatic drugs. These include inhibitors of signal transduction, cyclin-dependent kinase, angiogenesis, and matrix metalloproteinases. Other new therapies include gene therapy and immunotherapy. There are multiple steps in the signal transduction cascade. Growth factor binds to its cognate receptor tyrosine kinase and the phosphotyrosines on the receptor serve as attachment sites for substrates or adapter molecules. Grb2 functions by directly coupling activated receptor tyrosine kinases to the Ras-activating nucleotide exchange factor SOS. Activation of Ras or Ras family members leads to activation of the mitogen activated protein kinase (MAPK) cascade pathway. This has been implicated as a necessary component of intracellular signaling to elicit a range of cellular responses including mitogenesis, differentiation, and cell survival. We introduce signal transduction inhibitors including, Ras inhibitors, protein kinase C inhibitors, and MAPK cascade inhibitors. Recently, these drugs have been used in clinical trials and some of them have an antitumor effect. In the near future these drugs may play an important role in cancer chemotherapy. However, these drugs are thought to induce a non-cytotoxic effect different from cytotoxic drugs. Therefore correct and efficient clinical evaluation of these drugs is needed. We look forward to developments from future research on signal transduction inhibitors. PMID- 11383207 TI - [Tyrosine kinase inhibitor--hematological malignancies]. AB - STI571 selectively inhibits the ABL-tyrosine kinase, the activity of which is activated by the formation of chimeric BCR/ABL. A phase I study in the USA showed STI571 to be remarkably effective in cases of interferon-refractory chronic myeloid leukemia, with almost no adverse effects. STI571 may become the first choice drug prior to stem cell transplantation and interferon treatment. PMID- 11383208 TI - [Tyrosine kinase inhibitors--solid cancers]. AB - Tyrosine kinase inhibitors have drawn the most attention in recent years as molecular target agents for cancer treatment. The reason for this can only be the dramatic antitumor effects shown in early clinical trials against small cell cancer and chronic myeloid leukemia by the EGFR tyrosine kinase inhibitor, ZD1839, and the BCR-Abl tyrosine kinase inhibitor, STI-571, respectively. Various hypotheses were advanced in the preliminary stages of the clinical development of such molecular target agents: "They only prevent cancer cell proliferation and have no killer cell activity; they are extremely weak, and can not be expected to reduce tumors at the clinical level. "Or:" A long time is required before their physiological activity will be expressed in an antitumor effect." However, with non-small cell lung cancer, the most difficult tumor among solid cancers for an anticancer agent to be effective, not only was ZD1839 effective, but showed clear effectiveness in combination chemotherapy in the pretreatment stage. Moreover, the time for the expression of its tumor reduction effect was virtually the same as with conventional anticancer drugs, and its effectiveness proved to last longer after its initial expression. ZD1839 has succeeded in remaking the very image of molecular target agents for cancer treatment. In what follows, we focus mainly on the EGFR tyrosine kinase inhibitor, ZD1839. PMID- 11383209 TI - [Telomerase inhibitor]. AB - Telomerase is a ribonucleprotein enzyme that extends telomeric repeats at the end of chromosome DNA. Telomere maintenance by telomerase leads to cellular immortality and carcinogenesis. Telomerase is activated in most malignant tumors while it is usually repressed in normal somati tissues, and telomerase activation is thought to be a critical step for cancer progression. These findings suggest that telomerase is a good target for cancer gene therapy. A variety of molecular technique have been used to inhibit telomerase activity in cancer cells. In this review, I introduce the representative methods which efficently inhibit telomerase activity and may have a potential for clinical application. PMID- 11383210 TI - [Platinum compounds in cancer therapy--past, present, and future]. AB - Platinum cytotoxics play an important role globally in the management of solid tumours. Cisplatin sets the standard for efficacy in both regions with careful administration to reduce nephrotoxicity. Carboplatin is associated with neurotoxicity, but has become the leading product in the US due largely to the easier to manage toxicity profile. Both agents have been widely used in both registered and non registered indications and are frequently combined with other cytotoxics. In Japan, cisplatin has been used successfully at low doses in combination with 5-FU based regimens and appears to achieve a synergistic effect, but controlled data are not yet available. More recently oxaliplatin (Europe) and nedaplatin (in Japan) have been introduced, but their clinical roles in therapy have yet to be established. One of the limiting features of the first generation of platinum compounds is that a significant proportion of tumours develop cross resistance to platins due to either changes in uptake or excretion, intracellular detoxification or accelerated DNA repair. The forum discussed the possibility for the development of better new platinum compounds, A new platin agent which had lower toxicity and higher efficacy across a wide range of cancers without the development of resistance would be a significant step forward. If the tolerability profile was suitable, an oral formulation may improve the quality of life for patients but this must not be at the expense of efficacy. Even after the introduction of new target based drugs, platinum cytotoxics are likely to be used to reduce the tumour mass and in some cases can be expected to potentiate the effects of the new agents. In preclinical studies, ZD0473 has been shown to by pass some major mechanisms of resistance and has the potential to achieve these objectives and is now being evaluated in clinical studies in both Japan and the West. PMID- 11383211 TI - [Efficacy of docetaxel for anthracycline-resistant metastatic breast cancer]. AB - From June 1997 through December 1999, we treated twenty-two metastatic breast cancer patients with Docetaxel (60 mg/m2 administered intravenously every four weeks). All patients had received prior chemotherapy including anthracycline, and also another agent such as MMC-VDS, HDMTX-5-FU or CPT-11. One CR, 5 PR, and 4 Long NC were achieved. Grade 3/4 leukocytopenia occurred in 16 patients. Using the Kaplan Meier method, the median time to progression was 179 days, and the median survival time was 369 days. No serious adverse effects were observed. In this study Docetaxel seems to have shown significant activity, even for poor candidates, with intensive prior therapy. PMID- 11383212 TI - [Weekly paclitaxel and cisplatin in recurrent ovarian cancer]. AB - This study was performed to assess the feasibility of weekly paclitaxel (TXL) and cisplatin (CDDP) in patients with recurrent ovarian cancer. Ten of eleven patients experienced recurrence after more than 6 months after first line CDDP based chemotherapy. TXL and CDDP were given at initial doses of 60 mg/m2 and 30 mg/m2 on days 1, 8, and 15 in 2 patients and an increase in the respective dose level was planned to 60/35 in 5 patients, 70/35 in 2 patients, and 70/40 in 2 patients. Toxicities were well tolerated. None of the patients suffered from neurotoxicity or myalgia of more than grade 2. Gastrointestinal disorder was recognized as grade 1-2, and grade 3-4 hematological toxicity included leucocytopenia (64%), anemia (36%), and thrombocytopenia (9%). We set the recommended dose of TXL at 70 mg/m2 and that of CDDP at 35 mg/m2, considering toxicity and performed planned schedule. Of eleven patients, nine were assessable by computed tomographic scan. The overall response rate was 67% (CR: 1, PR: 5, NC: 1, PD: 2). One of two patients with standard TXL/CDDP therapy showed PR by switching to a weekly schedule. The median follow-up duration was 490 days and the median response duration was 371 days. From the results presented here, it is suggested that this regimen with increased DI might be quite effective and well tolerated in patients who experience relapse after CDDP-based chemotherapy. PMID- 11383213 TI - [Significance of pyrimidine nucleoside phosphorylase (PyNPase) and dihydropyrimidine dehydrogenase (DPD) in advanced gastric cancers]. AB - We measured pyrimidine nucleoside phosphorylase (PyNPase), a known angiogenetic factor, and dihydropyrimidine dehydrogenase (DPD) in advanced gastric cancers. PyNPase was expressed in cytoplasm of the cancer cells and surrounding interstitial cells. The levels of PyNPase and DPD were significantly higher in cancer tissue. With respect to tumor factors, the level of PyNPase was significantly higher in cases positive for venous invasion. We divided patients into two groups, with high and low activities of IAP and MMP-9. The level of PyNPase was significantly higher in the high IAP activity group. A correlation was suggested between the level of PyNPase and the activity of IAP. 5'-Deoxy-5 fluorouridine (5'-DFUR) is transformed into 5-FU by PyNPase and manifests antitumor effects. DPD is a rate-limiting enzyme in the process of degradation of 5-FU. In the present study, the level of PyNPase/DPD was significantly higher in cancer tissue. PyNPase/DPD suggests not only the malignant potential of the tumor but also the efficiency of chemotherapy using 5-FU, especially 5'-DFUR. PMID- 11383214 TI - [Relationship between the anti-metastatic effect of UFT and in vitro chemosensitivity to 5-FU in metastatic tumors from orthotopic implanted colon cancer in nude rats]. AB - We have investigated the correlation between the in vitro chemosensitivity to 5 FU, measured using the collagen gel droplet embedded culture drug sensitivity test (CD-DST), and the anti-tumor effect of UFT, a prodrug of 5-FU, in metastatic tumors from orthotopic implanted colon cancer in nude rats. Human colon cancer cells (KM12SM) were injected into the cecal wall of the nude rats. Five weeks later, the implanted cecal tumors were removed. Oral UFT (a daily dose of 30 mg/kg) was administered postoperatively for four weeks. After the UFT administration period, the lung and lymph nodes were analyzed macroscopically and microscopically. In vitro chemosensitivity to 5-FU in the lung and lymph node metastases was tested using CD-DST, and the enzymatic activities of thymidine synthetase (TS) and dihydropyrimidine dehydrogenase (DPD) in the lung and lymph node metastases were measured. A daily administration of UFT produced an inhibitory effect on lung metastasis compared with the control group. However, there was no difference in the frequency of lymph node metastasis. The inhibition rate produced by 5-FU in CD-DST was significantly higher for lung metastases than for lymph node metastases. There was no difference in the TS and DPD activities between the metastatic tumoral tissues. These results suggest that the organ specificity of the anti-tumor effects of UFT on colon metastases may be determined by CD-DST of 5-FU for individual tumors. The TS and DPD activity in the tumoral tissues may not affect the organ specificity of the anti-tumor effect of UFT on colon metastases. PMID- 11383215 TI - [Correlation between 5-fluorouracil (5-FU) sensitivity as measured by collagen gel droplet embedded culture drug sensitivity test (CD-DST) and expression of orotate phosphoribosyl transferase (OPRT), thymidylate synthase (TS), and dihydropyrimidine dehydrogenase (DPD) in colorectal cancer]. AB - The levels of OPRT, DPD, and TS were determined in colorectal cancer tissue specimens, and 5-FU sensitivity was measured by CD-DST. The correlation between enzyme activity and 5-FU sensitivity was then studied. Six patients with colorectal carcinoma who had undergone surgical resection in our institution between May and August 2000 were studied. The CD-DST method was used to measure the sensitivity to 5-FU under three sets of conditions: 0.2 microgram/ml x 5 days (A), 1.0 microgram/ml x 1 day (B), and 10.0 micrograms/ml x 3 h (C). The coefficients of correlation of tumor sensitivity to 5-FU and OPRT activity were A: 0.8246, B: 0.7670, and C: 0.7856, and to DPD activity were A: 0.2525, B: 0.3928, and C: 0.4337, while the coefficients of correlation to TS enzyme levels were A: -0.5240, B: -0.4770, and C: -0.6131. These findings demonstrate a high degree of correlation between OPRT activity at the tumor site and tumor sensitivity to 5-FU under a variety of conditions, suggesting that OPRT activity can be a useful indicator in predicting the anti-tumor effectiveness of 5-FU for a specific tumor. PMID- 11383216 TI - [Fundamental study on the sensitivity test inhibiting fibroblast proliferation]. AB - Although many kinds of chemosensitivity test are available for the selection of suitable anticancer agents, the results of in vitro tests against solid tumors have been generally influenced by mixed in fibroblasts, as have SDI tests. We demonstrated that the influence of fibroblasts can be excluded by performing the SDI test on an agar layer, thus significantly inhibiting the growth of fibroblasts in the liquid top layer. The sensitivity of the agar SDI test was determined on the 3-4th day after incubation because cell proliferation tended to be delayed, and there were also somewhat higher cell counts of about 40,000 per well. The inhibition indices with and without agar in vitro were the same, showing no significant differences. With nude mice, the transplanted tumor index value of the agar SDI test is higher than that of SDI, and approaches in value the SDI of a pure tumor cell, which means that the sensitivity of a solid tumor might be more accurately shown by an agar SDI test than by an SDI test. PMID- 11383217 TI - [Three cases of advanced gastric cancer resected after successful treatment with the novel oral anticancer drug TS-1]. AB - We operated on 3 patients with advanced gastric cancer after successful treatment with novel oral 5-fluorouracil derivatives (TS-1). In all 3 patients, gastrointestinal fiberscopy revealed a considerable reduction in the tumor after oral administration of TS-1 in our clinic. Case 1 underwent surgery after an interval of 1 day following oral administration of TS-1 for 18 days. Pathologically, no cancer cells were found in the resected stomach and a few cancer cells were found only in resected lymph nodes. In case 2, who underwent surgery after an interval of 7 days following oral administration of TS-1 for 14 days, a few cancer cells were found only in the submucosal layer of the stomach and no viable cancer cells remained in the metastasized lymph nodes. In case 3, who underwent surgery after an interval of 11 days following administration of TS 1 for 26 days, scattered cancer cells, mostly fibrotic, were found as far as the subserosal layer. The pathological effectiveness of chemotherapy was grade 2 or 3 in each case. Case 2 and 3 followed a satisfactory postoperative course; however, case 1 suddenly had endotoxinemia on the 7th postoperative day. Although the patient had temporary multiple organ failure, he recovered eventually. All 3 patients have been well without any signs of recurrence for more than a year after surgery. PMID- 11383218 TI - [Two cases of alpha-fetoprotein producing gastric cancer, showing marked response to continuous hepatic arterial infusion (HAI) chemotherapy with adriamycin]. AB - We report two cases of alpha-fetoprotein producing gastric cancer (AFPGC) with multiple liver metastases showing marked response to continuous HAI chemotherapy with adriamycin (ADM). In both cases, 5-FU 500 mg/body/day and Leucovorin (LV) 30 mg/body/day was infused continuously for 7 days and ADM 30 or 60 mg/body/day was infused continuously for 4 hours on day 7 as preoperative HAI chemotherapy. The primary gastric lesions were reduced and became resectable. After gastrectomy, they were treated with 4-hour continuous HAI of ADM 30 or 60 mg/body with or without 5-FU and LV once a week several times in our outpatient clinic. After these treatments, the multiple liver metastases were reduced remarkably, with a marked decrease of serum AFP levels. During these treatments, neither patient showed remarkable side effects, so they could work as before. This frequently low dose ADM administration resulted in a high local response without severe side effects. PMID- 11383219 TI - [Gastric cancer associated with dermatomyositis accompanied by photoallergy]. AB - A 62-year-old man was admitted to the hospital because of a 5-month history of edematous erythema and itching on the face, upper chest, and upper extremities. The symptoms developed immediately after bathing in the sea. Dermatomyositis associated with photoallergy was diagnosed by skin and muscle biopsy. A search for malignancy revealed Borrmann 3 gastric cancer, and subtotal gastrectomy was performed. He has done well for 3 years and 8 months after the operation, but there has been no remission in the symptoms of dermatomyositis. Dermatomyositis associated with photoallergy has a higher incidence of complications with malignant disease than ordinary dermatomyositis. PMID- 11383220 TI - [Successful resection of multiple liver metastases from rectal cancer following initial treatment using hepatic arterial infusion chemotherapy and radiotherapy]. AB - A 69-year-old-man was referred to our hospital because of rectal cancer with multiple liver metastases. He was initially treated by hepatic arterial chemotherapy using an infusion reservoir (HACR) and radiotherapy for the rectal cancer. An abdomino-perineal resection and extended left lobectomy of the liver were performed and resulted in a reduction in size of the liver tumor. He was diagnosed as having a recurrent liver metastasis (S7) at 3 months after the operation, and was retreated by HACR in the outpatient clinic. A partial hepatectomy was reperformed at 6 months after the operation. Adjuvant hepatic arterial infusion chemotherapy (HAIC) was performed on an outpatient basis and the patient is doing well without recurrence or relapse. Preoperative arterial chemotherapy for metastatic liver tumor may be of some benefit for certain patients with far advanced colorectal carcinoma. PMID- 11383221 TI - [Non-small-cell lung cancer (NSCLC) in an elderly patient--a case of squamous cell carcinoma successfully treated with chemotherapy using vinorelbine]. AB - We report an elderly patient with squamous cell carcinoma who was successfully treated with chemotherapy using vinorelbine. A 76-year-old man was referred to our hospital for evaluation of a nodular shadow in the left lung. Chest CT scam showed a 3-cm tumor shadow in left S9 and a 1-cm small nodule in right S2. Transbronchial lung biopsy yielded a diagnosis of squamous cell carcinoma. The clinical stage was IV (cT2N2M1). The patient first underwent chemotherapy consisting of cisplatin (CDDP) 80 mg/m2 on day 1 and vinorelbine (VNB) 20 mg/m2 on days 1, 8, and 15, which generated tumor shrinkage of 48% as well as transient elevation of grade 1 in serum creatinine. The 2 cycles of chemotherapy using vinorelbine only (VNB 20 mg/m2 on days 1, 8, 15) produced a tumor reduction of 70% with grade-1 decrease of granulocytes. The low grade of toxicity enabled us to treat the patient in our outpatient office for the second cycle of the regimen. This case suggests that chemotherapy using low-dose vinorelbine might be suitable to treat elderly patients with NSCLC in outpatient settings. PMID- 11383222 TI - [A modified administrated schedule for combination therapy with irinotecan and cisplatin as a neoadjuvant chemotherapy in locally advanced cervical cancer--a report of 2 cases as a pilot study]. AB - To shorten the treatment term of neoadjuvant chemotherapy (NAC) for locally advanced cervical cancer, a combination of irinotecan (CPT-11) and cisplatin was administered on a modified administrated schedule for 2 eligible patients as a pilot study. CPT-11 70 mg/m2 was administered (div) on days 1 and 8, followed by cisplatin 70 mg/m2 given (div) on day 1. Treatment was repeated every 3 weeks for a total of two cycles. Both patients showed a complete clinical response. No severer toxicities were observed than with the usual regimen, and both cases could undergo radical hysterectomy after NAC. The results suggest that this modified regimen of combination of CPT-11 and cisplatin can be effective as an NAC in cases of locally advanced cervical cancer, especially in terms of shortening treatment term. This regimen is worthy of further study. PMID- 11383223 TI - [Non-small cell lung cancer with liver metastasis responsive to gemcitabine--a case report]. AB - A 56-year-old male patient underwent a right upper lobectomy and lymph node dissection for non-small cell lung cancer in March 1994. Multiple lung metastases in the right lung were found 45 months after the operation, and chemotherapy with docetaxel was administered. A liver metastasis was detected 11 months later, and it was refractory to docetaxel. Therefore, the patient was treated with cisplatin, mitomycin C and vinorelbine, which resulted in no change to the liver metastasis. He was next treated with gemcitabine, which resulted in a partial response of the liver metastasis. The adverse effects of gemcitabine were Grade 3 thrombocytopenia and Grade 2 neutropenia. The response duration for gemcitabine therapy was three months. PMID- 11383224 TI - [Application of population pharmacokinetic approach to clinical evaluation of anticancer agents]. AB - Population pharmacokinetics deals with the typical profiles and the inter- and intra-individual variability in the target patient population to which the drug is applied. It also describes factors that can affect the inter-individual variability in pharmacokinetics, including physiological, pathological, genetic, and external factors. The sample population is the actual patients with a variety of backgrounds, which enables us to analyze the influences of several factors such as severity of illness, advanced age, childhood, and renal or hepatic dysfunction, and also to identify the special populations where dose adjustment is needed. Pharmacokinetic and pharmacodynamic information are useful to a rational dosage regimen. The findings obtained by the population pharmacokinetic and pharmacodynamic analysis, provide an advice about whether a dosage regimen should be individualized in all patients or in identified special populations, together with how to adjust the dose. The study design for population pharmacokinetic analysis is called "pharmacokinetic screen", where drug concentration data are collected from a large number of patients while only a few blood samples are taken from individual patients. The population pharmacokinetic approach is useful not only for establishing the rationale dosage regimen but also for international exchange of clinical data in the global drug development. PMID- 11383225 TI - [Tumor markers for hepatocellular carcinoma]. AB - The diagnosis of hepatocellular carcinoma (HCC) is based mainly on serological tumor markers, such as alpha-fetoprotein, L3% fraction thereof and PIVKA-II, and imaging modalities. These are not correlated but are complementary. Hence, a combination designed to take advantage of the characteristics of each needs to be worked out. First, it is necessary to identify the patients at high risk for developing HCC, such as those with chronic hepatitis or liver cirrhosis, and in the follow-up conduct regular check-ups for serological tumor markers. Those testing positive for any marker are at the highest risk for developing HCC, even when imaging fails to disclose any space-occupying lesions. Following these high risk patients, in concert with imaging, enables accurate evaluation of the efficacy of therapies for HCC. Since serological tumor markers can signal the development of HCC earlier than any other laboratory test, they offer an excellent means of identifying relapsing HCC. Equally important in the management of patients with HCC are biological indicators for malignancy, selection of therapeutic interventions and prediction of the outcome. PMID- 11383226 TI - Allogeneic haematopoietic stem cell transplantation with reduced intensity conditioning regimens ("minitransplants"). AB - During the recent years it became evident that allogeneic haematopoietic stem cell transplantation with reduced intensity conditioning is feasible. Though less toxic than conventional stem cell grafting severe adverse effects have been reported and are not uncommon. Due to the still short observation periods realistic outcome estimations cannot be made yet. Nevertheless allografting with reduced intensity conditioning is a promising new therapeutic option and hopefully its true position in the treatment of haematologic and oncologic diseases will become more clear in the next future. PMID- 11383227 TI - Treatment of congenital aortic valve stenosis: impact of the Ross operation. AB - GOAL: To evaluate the impact of the Ross operation, recently (1997) introduced in our unit, for the treatment of patients with congenital aortic valve stenosis. METHODS: The period from January 1997 to December 2000 was compared with the previous 5 years (1992-96). Thirty-seven children (< 16 yrs) and 49 young adults (16-50 yrs) with congenital aortic valve stenosis underwent one of these treatments: percutaneous balloon dilatation (PBD), aortic valve commissurotomy, aortic valve replacement and the Ross operation. The Ross operation was performed in 16 patients, mean age 24.5 yrs (range 9-46 yrs) with a bicuspid stenotic aortic valve, 7/10 adults with calcifications, 2/10 adults with previous aortic valve commissurotomy, 4/6 children with aortic regurgitation following PBD, and 1/6 children who had had a previous aortic valve replacement with a prosthetic valve and aortic root enlargement. RESULTS: PBD was followed by death in two neonates (fibroelastosis); all other children survived PBD. Although there were no deaths, PBD in adults was recently abandoned, owing to unfavourable results. Aortic valve commissurotomy showed good results in children (no deaths). Aortic valve replacement, although associated with good results (no deaths), has been recently abandoned in children in favour of the Ross operation. Over a mean follow-up of 16 months (2-40 months) all patients are asymptomatic following Ross operation, with no echocardiographic evidence of aortic valve regurgitation in 10/16 patients and with trivial regurgitation in 6/16 patients. CONCLUSIONS: The approach now for children and young adults with congenital aortic valve stenosis should be as follows: (1) PBD is the first choice in neonates and infants; (2) Aortic valve commissurotomy is the first choice for children, neonates and infants after failed PBD; (3) The Ross operation is increasingly used in children after failed PBD and in young adults, even with a calcified aortic valve. PMID- 11383228 TI - Risk free simultaneous prenatal identification of fetal Rhesus D status and sex by multiplex real-time PCR using cell free fetal DNA in maternal plasma. AB - QUESTIONS UNDER STUDY: Pregnancies with a Rhesus constellation still present a considerable obstetric problem. In addition, pregnancies with male Rhesus D fetuses are more severely affected by haemolytic disease of the newborn, requiring more transfusions in utero and having a three fold higher mortality than female Rhesus D fetuses. Furthermore, almost 150 X-linked genetic deficiencies have now been characterised, increasing the need for prenatal sex determination in pregnancies at risk for such a disorder. In order examine these two important fetal loci in a risk free manner, we have developed a novel multiplex real-time PCR assay for the analysis of extracellular fetal DNA in maternal plasma. METHODS: Cell free DNA was isolated from 34 maternal plasma samples and examined by a multiplex real-time PCR assay for the Rhesus D gene and the SRY locus on the Y chromosome. RESULTS: Our study showed that we were able to genotype 12/13 Rhesus D males correctly. All 5 Rhesus d males were correctly identified. In addition a 100% concordance was found in the 16 samples obtained from pregnancies with female Rhesus D or Rhesus d fetuses. CONCLUSIONS: By developing a novel multiplex real-time PCR assay we present the first report describing the determination of multiple fetal loci from cell free DNA in maternal plasma by these means. As this assay is suitable for automation, our data, therefore, suggest that such assays provide a good basis for the clinical examination of multiple fetal loci, in particular Rhesus D status or fetal sex, and can be performed effectively using real-time multiplex PCR assays. PMID- 11383229 TI - Therapeutic efficacy of selective nerve root blocks in the treatment of lumbar radicular leg pain. AB - The objective of this study was to investigate the clinical effectiveness of nerve root blocks (i.e., periradicular injection of bupivacaine and triamcinolone) for lumbar monoradiculopathy in patients with a mild neurological deficit. We have retrospectively analysed 30 patients (29-82 years) with a minor sensory/motor deficit and an unequivocal MRI finding (20 disc herniations, 10 foraminal stenoses) treated with a selective nerve root block. Based on the clinical and imaging findings, surgery (decompression of the nerve root) was justifiable in all cases. Twenty-six patients (87%) had rapid (1-4 days) and substantial regression of pain, five required a repeat injection. 60% of the patients with disc herniation or foraminal stenosis had permanent resolution of pain, so that an operation was avoided over an average of 16 months (6-23 months) follow-up. Nerve root blocks are very effective in the non-operative treatment of minor monoradiculopathy and should be recommended as the initial treatment of choice for this condition. PMID- 11383230 TI - Reactive hypoglycaemia due to late dumping syndrome: successful treatment with acarbose. AB - Reactive hypoglycaemia is a rare disease which occurs postprandially in everyday life involving blood glucose levels below 2.5 to 2.8 mmol/l. We report on a 66 year-old patient who developed symptomatic reactive hypoglycaemia due to late dumping syndrome 10 years after oesophagectomy with cervical anastomosis. A 75 g sucrose load revealed a plasma glucose level of 9.4 mmol/l after one hour, followed by symptomatic hypoglycaemia with a plasma glucose level of 1.8 mmol/l after three hours. Concomitantly, high concentrations of insulin (3216 pmol/l at a glucose level of 9.4 mmol/l and 335 pmol/l at a glucose level of 1.8 mmol/l) and glucagon-like peptide 1 (GLP-1) (375 pmol/l at a glucose level of 9.4 mmol/l and 85 pmol/l at a glucose level of 1.8 mmol/l) were measured. While the patient was under treatment with acarbose, another sucrose load did not provoke symptomatic hypoglycaemia (plasma glucose nadir of 4.6 mmol/l after two hours). Insulin and GLP-1 levels increased much less, to peak levels of 375 pmol/l and 75 pmol/l respectively, after one hour when plasma glucose was 6.8 mmol/l. We conclude that in patients with reactive hypoglycaemia due to gastrointestinal surgery, acarbose decreases rapid glucose absorption associated with hyperglycaemia and GLP-1 secretion, and thus diminishes excessive insulin release. Acarbose is therefore a successful treatment modality for reactive hypoglycaemia due to late dumping syndrome. PMID- 11383231 TI - Therapeutic inadequacy in spite of bioequivalency on replacing Fluctine with Fluocim. PMID- 11383232 TI - [Root caries in patients in periodontal follow-up care. Prevalence and risk factors]. AB - The aim of this cross-sectional study was to investigate the prevalence and several risk indicators of root caries in 45 periodontal maintenance patients, who had been actively treated for adult periodontitis 11-22 years ago. These patients were part of a routine 3-6 monthly maintenance schedule. Active and inactive root caries and root fillings were recorded, as well as coronal caries experience. Plaque and bleeding scores, number of exposed root surfaces, rate of saliva secretion, saliva buffer capacity, S. mutans counts and Lactobacilli were also scored. From the total of 45 subjects, 37 patients (82%) showed root lesions (root caries and/or fillings), while only 8 patients were free of any root lesions. On average, there were 4.3 lesions per patient (range 0-19) in the present study. Of all damaged root surfaces, 9% were active lesions, mostly located on mandibular teeth at lingual and vestibular sites; 40% were inactive lesions often detected at vestibular sites. The remaining damaged root surfaces (51%) were restored; they were equally divided over both jaws. A higher number of root lesions was observed in those patients with > 106 S. mutans/ml saliva. Although the actual number of lesions per patient was low in relation to the large number of sites with gingival recession, the results from this cross sectional study in periodontal maintenance patients indicate that: root caries can be regarded as a complication in periodontal maintenance patients, that the individual number of root lesions correlate with individual dental plaque score, that a high number of root lesions is associated with counts of salivary S. mutans, and that no relation between root caries and coronal caries experience, salivary secretion rate or salivary buffering capacity seems present. Therefore, repeated oral hygiene instructions and adjunctive preventive measures including diet counseling and fluoride rinses, as well as fluoride and chlorhexidine varnishes, should be advocated in high-risk patients. PMID- 11383233 TI - [Treatment protocol for craniomandibular dysfunction 2. Treatment]. AB - In two essays the treatment strategy of the department of Oral Function of the Academic Center of Dentistry Amsterdam for treating craniomandibular disorder (CMD) patients is described. In this second essay a description is given of several dental, physiotherapeutical and psychological treatment modalities for CMD. Then treatment strategies for the different categories of CMD are described. It is also indicated which aspects of the treatment strategy are based upon 'evidence based care' and which aspects are more based upon principles of 'common sense' and 'clinical prudence'. PMID- 11383234 TI - [Oral aspects of Crohn's disease]. AB - Crohn's disease is a chronic inflammatory disease, which may involve any part of the gastrointestinal tract, including the oral cavity. This review discusses possible causes, consequences and medical treatment of Crohn's disease. In addition, an overview of the oral manifestations of this disease and the implications for dental treatment are presented. PMID- 11383235 TI - [Corticotomy of the upper jaw: a different view]. PMID- 11383236 TI - [Developments in dentistry in the twentieth century 8. Endodontics]. PMID- 11383237 TI - [Bite, swallow, away?]. PMID- 11383238 TI - [Human genome 2]. PMID- 11383239 TI - [Center for smell and taste disorders]. PMID- 11383240 TI - [Taking impressions of a somewhat resorbed jaw]. PMID- 11383241 TI - Expectant management of prostate cancer. AB - OBJECTIVES: To discuss the role of expectant management in the treatment of clinically localized prostate cancer. DATA SOURCES: Published research and review articles, textbooks, and pending research publications. CONCLUSIONS: Expectant management is a viable option for the treatment of clinically localized prostate cancer in carefully selected men. IMPLICATIONS FOR NURSING PRACTICE: Nursing personnel along with physicians must work together to develop coping strategies for these men to deal with the continual uncertainty of this treatment program. PMID- 11383243 TI - Treatment of advanced prostate cancer. AB - OBJECTIVES: To review the management of advanced prostate cancer, including symptom management, hormonal therapy, and the use of chemotherapy. DATA SOURCES: Published articles and book chapters on advanced prostate cancer. CONCLUSIONS: Advanced prostate cancer is a common problem that has a significant impact on the patient's quality of life. Multiple complications may develop during the course of the disease. Treatment may include local and systemic approaches. Advances in disease treatment include hormonal therapies and chemotherapy. Additional research is needed to determine the optimal treatment for these men. IMPLICATIONS FOR NURSING PRACTICE: Patient education is crucial to the management of advanced prostate cancer in all practice settings. Early and prompt recognition of disease complications will enhance the patient's quality of life. PMID- 11383242 TI - Decision making and prostate cancer treatment selection: a review. AB - OBJECTIVES: To review the issues surrounding decision making regarding prostate cancer treatment. DATA SOURCES: Journal articles, abstracts, and selected findings from one prospective grounded theory research study. CONCLUSIONS: Multiple factors are involved in the selection of prostate cancer treatment, such as individual disease-specific data, age, personal values, vicarious and personal cancer experiences, and the physician-patient relationship. IMPLICATIONS FOR NURSING PRACTICE: Accurate information regarding each treatment option delivered by experts in their respective fields is necessary for informed decision making. A more multidisciplinary approach regarding discussion of treatment options is needed. Nurses play a pivotal role in facilitating discussion among family members and assisting patients to make a decision that balances their personal values with accurate information regarding treatment outcomes. PMID- 11383244 TI - Psychosocial and educational intervention trials in prostate cancer. AB - OBJECTIVES: To describe the nature and focus of published educational intervention trials in prostate cancer in the last 10 years and to examine the state of the research in this particular area of knowledge. DATA SOURCES: Health science and psychosocial electronic databases were searched for studies reporting trials of educational or psychosocial interventions for men with prostate cancer and for the partners or families of these men. CONCLUSIONS: Interventions were characterized by the aspect of the illness trajectory that was targeted. The majority of the studies focused on managing the disease, treatments, and side effects. IMPLICATIONS FOR NURSING PRACTICE: Psychosocial stressors and uncertainty are pervasive throughout the prostate cancer trajectory. Knowledge of psychosocial and educational interventions that have been tested is critical for nurses working with these patients and their families. PMID- 11383245 TI - Quality of life in patients with prostate cancer. AB - OBJECTIVES: Quality of life (QOL) issues are important for patients with prostate cancer because side effects from treatment are substantial, while the disease itself may be indolent. This article reviews prostate cancer QOL studies. DATA SOURCES: Selected studies published on QOL in prostate cancer using validated patient-assessed tools from the last 5 years. CONCLUSIONS: Prostate cancer treatments are associated with side effects: prostatectomy has more urinary and sexual side effects, while external radiation therapy has more bowel symptoms. Side effects are not highly correlated with overall QOL. IMPLICATIONS FOR NURSING PRACTICE: Patients must be made aware of potential gains in life expectancy as well as side effects of treatments to make informed decisions about treatment. PMID- 11383246 TI - The epidemiology of prostate cancer in the United States. AB - OBJECTIVES: To provide a review of prostate cancer epidemiology. DATA SOURCES: Journal articles and the Surveillance, Epidemiology, and End Results database. CONCLUSIONS: Numerous risk factors for prostate cancer have been identified. Incidence and mortality rates are higher in certain countries and among racial ethnic groups. Environmental and dietary influences are likely to be significant causes of prostate cancer. Distinguishing those cancers that are clinically significant from those that are not remains a central concern in prostate cancer research. IMPLICATIONS FOR NURSING PRACTICE: Knowledge of epidemiologic patterns and trends will assist nurses in recognising persons at high risk for the development of prostate cancer and should result in tailored educational interventions. PMID- 11383247 TI - The debate about prostate cancer screening: what nurses need to know. AB - OBJECTIVES: Special challenges and unique opportunities for nurses in the 21st century related to prostate cancer screening are reviewed. DATA SOURCES: Current health care literature pertaining to prostate cancer screening issues. CONCLUSIONS: Decisions that weigh the immediate risks of incontinence and erectile dysfunction against the long-term potential risk of death from advanced cancer must be made with conflicting values and incomplete data. IMPLICATIONS FOR NURSING PRACTICE: With their expertise in patient education nurses are in a unique position to communicate the risks and benefits of prostate cancer screening in a manner in which patients can understand. A sample nursing care plan is presented for shared decision making. PMID- 11383248 TI - The role of surgery in the treatment of prostate cancer. AB - OBJECTIVES: To describe the role of surgery in the treatment of organ-confined prostate cancer and advanced prostate cancer. DATA SOURCES: Journals, textbooks, and personal communication. CONCLUSIONS: Radical prostatectomy plays a possible role in curing those patients with disease confined to the prostate. Bilateral orchiectomy plays a palliative role for patients with advanced disease. IMPLICATIONS FOR NURSING PRACTICE: This article provides nurses with the knowledge needed to teach patients about how the surgery is performed, the associated complications, and the likelihood of cancer control. PMID- 11383249 TI - Radiation therapy for prostate cancer. AB - OBJECTIVES: To review the advances in radiation therapy for prostate cancer and the nursing care of patients with prostate cancer. DATA SOURCES: Peer-reviewed journal articles, including research studies and review articles. CONCLUSIONS: Radiation therapy is used to cure early stage prostate cancer, control locally advanced disease, and effectively palliate symptoms of metastasis. The three forms of treatment used include external beam radiation therapy, brachytherapy; and radiopharmaceutical treatments. IMPLICATIONS FOR NURSING PRACTICE: Nursing care of patients receiving radiation therapy for prostate cancer includes managing the symptoms associated with the disease and treatment, educating patients and families about self-care measures, and providing support throughout the course of the disease. PMID- 11383250 TI - [Team approach in dentistry]. PMID- 11383251 TI - [Team management of orofacial clefts]. AB - In the Netherlands 15 centres provide multidisciplinary care for cleft lip and palate patients. Usually the following disciplines participate in such teams: paediatrics, plastic and reconstructive surgery, orthodontics, genetics, social work or nursing, ENT, speech therapy, maxillofacial surgery, prosthetic dentistry, psychology and oral hygiene. An overview is given of the treatment protocol from birth until 20 years of age for a child with a complete UCLP or BCLP. It is concluded that properly designed prospective clinical trials are rare, resulting in a lack of evidence based care in the field of cleft lip and palate. Furthermore it should be investigated whether it is preferable to centralise the cleft care in less centres than the present 15 ones. PMID- 11383252 TI - [Team approach in maxillofacial oncology]. AB - The most frequent malignant tumour in the head and neck region this area is the squamous cell carcinoma of the upper aerodigestive tract. In the Netherlands the incidence of these tumours is low, about 2000 new cases a year. Multidisciplinary treatment of these tumours is mandatory because of the extension of the lesion in the area of other disciplines. The variety of diagnostic and therapeutic possibilities and the supportive care ask for a multidisciplinary approach. The low incidence of head and neck cancer in the Netherlands and the complexity of the treatment of these tumours were the reason to concentrate the care for these patients in multi disciplinary head and neck cancer centres. PMID- 11383253 TI - [Multidisciplinary management of trauma patients. Role of the maxillofacial surgeon]. AB - The approach of patients with multiple trauma is considered to be complicated. Many (non)medical disciplines are involved. This article describes the treatment in the early post-traumatic period and the organisation of the care in both the pre-hospital and hospital phase. Special attention is given to the treatment of patients with maxillofacial injuries. The task of the oral and maxillofacial surgeon in particular when dealing with facial injuries is described. PMID- 11383254 TI - [Osteotomy team]. AB - The way the surgical orthodontic team approach has been developed over the years is described in its historical context. Originally orthodontists and surgeons worked more or less separately from each other. Nowadays a team of specialists including psychologists takes care of the patient. The importance to balance the indications for surgical orthodontic treatment against the contra-indications is emphasized. PMID- 11383255 TI - [Diagnosis and treatment of orofacial pain]. AB - Management of orofacial pain should address factors that contribute to the continuation of the pain complaint. This requires elimination of etiologic factors, modification of nociceptive mechanisms, and support of adaptive mechanisms. In addition, methods that help the patient to cope with (remaining) complaints should be applied. A specific diagnosis of the pain problem is the basis for successful management. From the history and clinical examination it should become clear whether the pain problem is acute or chronic, whether its characteristics suggest a somatic or neuropathic basis, and whether the experienced pain is primary or secondary (i.e., hyperalgesia, referred pain). It is important to regard pain as a multidimensional phenomenon, in which psychosocial factors are involved. It is the responsibility of the dentist to judge as early as possible whether he/she is capable of managing the pain problem, or further consultation or referral is necessary for diagnosis and/or management. PMID- 11383256 TI - [Multidisciplinary diagnosis and treatment of craniomandibular disorders]. AB - Treatment of craniomandibular disorders (CMD) requires a multidisciplinary approach. The CMD-team of the Academic Centre for Dentistry Amsterdam (ACTA) therefore consists not only of specialists in CMD and orofacial pain, but also of physiotherapists and a psychologist. Possible CMD-patients are referred to this team by the dentist-general practitioner, either directly or upon request of a family physician or a medical specialist; the many rules of such referrals are outlined in the article. The CMD-team regularly consults other disciplines for diagnosis and/or treatment of their patients. For instance, internal referrals are sometimes made to the departments of Oral radiology, Endontology, Periodontology, Orthodontics, or Oral and maxillofacial surgery. External referrals to, for example, a speech therapist or a medical specialist (e.g., a pain specialist) are arranged by the family physician upon request of the CMD team. PMID- 11383257 TI - [Combined periodontal treatment with dental implants]. AB - Implant treatment of partial edentulous periodontal compromised patients have led to a change in indication and therapy. After an adequate periodontal treatment for most of the patients there are no obstructions for implant treatment. However untreated periodontal disease and refractory periodontitis patients are at risk for complications. The diagnosis, treatment planning, treatment and maintenance of periodontal compromised patients need a multidisciplinary approach. A regular and tight maintenance program is essential to keep the periodontal and peri implant tissues in a good condition. PMID- 11383258 TI - [Cooperation between the dentist and the dental technician]. AB - In prosthetic dentistry the final result is mainly determined by the co-operation within the team: the dentist, the implantologist, the dental technician and the patient. The technician is more and more involved in the composition and colouring of the dental restorations. In many laboratories a dental unit is available to maximise the co-operation and to involve the patient if necessary. A plea is made to develop protocols to improve the co-operation between dentist and dental technician. At the same time new communication techniques can be used such as digital photography. As the costs are often high a treatment plan and an estimate of expenditure are necessary. It is advised that the dentist asks the technician to submit an offer. The dentist should realise that patients, because of the high costs, often have great expectations of the final result, which not always can be fulfilled. PMID- 11383259 TI - [Hormone dysregulators. Pseudo-estrogens in dental composite resins and sealanta?]. AB - A number of polluting chemicals in the ecosystems must be characterized as hormone disruptors. Among others, male animals appear to become feminized by the action of the so-called pseudo-estrogens and under their influence mens' fertility is said to decrease. Composites and sealants based on Bis-GMA resin may contain bisphenol-A as an impurity and Bis-DMA, from which in saliva bisphenol-A will be formed by hydrolytic degradation. Therefore, in extreme circumstances a weak estrogenic effect is not impossible on the short-term. However, the amounts of these probably not very potent estrogenic compounds are small, thereby resulting in a tolerable risk on the short term. Long-term-effects and synergism with pseudo-estrogens from other sources prompt further studies in order to verify the safety of the Bis-GMA containing products. PMID- 11383260 TI - [Male fertility. Possibly affected by occupational exposure to mercury]. AB - In the last decades there is circumstantial evidence for an increased incidence of cryptorchidisme and hypospadia in new-born boys and an increased incidence of testicular cancer in adults. Moreover, there are indications that sperm quality is declining. Exposure to endocrine disrupters in utero and during adulthood is a possible explanation for the impairment of male reproductive health. Exposure to mercury may lead to deposition in the pituitary gland and the testicular tissues causing impairment of testicular function, especially spermatogenesis. Evidence based research is needed. PMID- 11383261 TI - [Oral health of community-living elderly. 1. Condition of teeth, use of professional dental care and oral hygiene habits]. AB - With the aim to get information on oral health, professional dental care and oral hygiene habits a study was performed on persons aged 60 up to 79 years in Haarlem, the Netherlands. After an interview an oral examination was performed, using a mirror and a pocket lamp. Of all persons approached, 376 (38%) participated in the clinical part of the study. In elderly persons with natural teeth (52% of all participants) the mean numbers of present teeth (third molars excluded), FT and obvious DT were 18.1, 10.8 and 0.7 respectively. The percentage of persons with natural teeth showed a strong relationship with level of education. A comparison with results of a study performed in 1986 showed that oral health in elderly persons is improved. Of all the elderly with natural teeth 91% visited their dentist periodically at least one time a year and 76% of them brushed their teeth at least two times a day. PMID- 11383262 TI - [Comparison of treatment options for myogenous temporomandibular dysfunction]. AB - A randomized clinical trial which included a follow-up of one year was applied to 118 myogenous TMD-patients. The therapies investigated were: physiotherapy of the masticatory system or splint therapy for patients without occlusal interferences, and occlusal adjustment therapy or the combination of splint and occlusal adjustment therapy for patients with pronounced occlusal interferences. Counseling, which yielded a reduction of 27% of the scored pain intensity, will most likely eliminate any further need for treatment of patients with a low level of myogenous TMD signs and symptoms. Otherwise, physiotherapy might be preferred as a starting option with respect to splint therapy because of 1. A similar efficacy; 2. A shorter treatment duration so that either chronic facial pain is earlier relieved or a patient can undergo a second type of therapy earlier; and 3. Lower costs. One third of the patients selected had pronounced occlusal interferences. Using stringent criteria it might be possible to apply occlusal adjustment therapy without involvement of splint therapy yielding a similar therapy efficacy and with advantages of a shorter treatment duration and lower costs. All types of therapy diminished not only facial pain but also pain of neck and shoulder areas. PMID- 11383263 TI - [Comment on 'Comparison of treatment options in myogenous temporomandibular dysfunction']. PMID- 11383264 TI - [Developments in dentistry in the twentieth century 9. Periodontology]. PMID- 11383265 TI - [Root (caries) and other statistical treatments in dentistry]. PMID- 11383266 TI - [Hypersalivation, lesions of oral mucosa and generalized fatigue]. AB - In case of a combination of hypersalivation and mucosal disease that rapidly progresses towards becoming indisposed, one has to consider an underlying general disease. In this paper a patient with these complaints, who visited the emergency department of the Groningen University Hospital, is described. The patient appeared to be suffering from a specific form of erythema multiforma, viz. the syndrome of Stevens-Johnson. PMID- 11383267 TI - [Schizophrenia 1]. PMID- 11383268 TI - [How much sodium hypochlorite should be used in root canal treatment?]. PMID- 11383269 TI - [Complaints against dentists]. AB - To get an impression of the claim-behaviour of patients of Dutch dentists, a qualitative survey was completed amongst eight out of the nine dental consultants of dental liability insurers in the Netherlands. After a literature-study and a test-interview, 11 questions were composed to get the information. It became apparent that the accused dentist is either the young, (too) enthusiastic dentist or the older one, who has lost interest in his job and follows no post academic education. Like in most western countries, according to literature, the most frequent topics patients complain about in the Netherlands are oral surgery, followed by fixed prosthetics, endodontology and recently periodontology. The main cause of the complaints is, according to the interviewed advisors, the lack of communication between the dentist and his patient. Moreover it became clear that dentists misuse their malpractice liability insurance as a second patient insurance. PMID- 11383270 TI - [Does history repeat itself?]. PMID- 11383271 TI - [Is the dentist presently no more qualified to construct a dental prosthesis?]. PMID- 11383272 TI - [Developments in dentistry in the twentieth century. 10. Infection prevention]. PMID- 11383273 TI - [Capacity dental care: short and long-term recommendations]. PMID- 11383274 TI - [Hyperparathyroidism with maxillo-mandibular deformation]. AB - Hyperparathyroidism is a condition due to an elevated secretion of parathormone. Oral manifestations are caused by the disturbance of the calcium-phosphate equilibrium which results in pathologic changes of the bones. A patient with secondary hyperparathyroidism and severe maxillary and mandibular deformations is described. PMID- 11383275 TI - [Schizophrenia 2]. PMID- 11383276 TI - [Assurance of original radiographs in the digital format]. PMID- 11383277 TI - [Clinical evaluation of the Biocomp-implant system. Results after 1,3 and 5 years in a general practice]. AB - A population of 107 consecutive patients was treated with a new HA-coated, screw type dental implant (n = 384 implants), the Biocomp implant. Treatment was performed in a general dental practice by one and the same dentist. The implants were utilized for various types of prosthetic constructions, sometimes in challenging conditions with respect to patient selection, jaw bone quantity and quality. A clinical and radiographical evaluation was performed after 1, 3 and 5 years. After 5 years, the chance on implant survival was 87.1% (standard error 3.4%). Implant loss occurred predominantly within the first 1.5 year after implant placement. Particularly short implants, placed in the atrofic maxilla, frequently failed. Biocomp implants that served as abutments for an overdenture in the edentulous mandible or as a single tooth replacement in the upper anterior region had an excellent prognosis. PMID- 11383278 TI - [Repair and revision 1. Repair or replacement of amalgam]. AB - Amalgam restorations may suffer from all sorts of shortcomings. Three options are available to the dental practitioner: replacement, repair or no repair. Minor shortcomings which do not cause any harm are better left untreated. Reasons for repair or replacement are: secondary caries, failing proximal contact, cervical overhanging, bulk fracture, fracture of adjacent tooth structure and postoperative sensibility. Repair will cause less iatrogenic damage than complete replacement, and is therefore preferred. Amalgam restorations can be repaired with composite resin (in conjunction with an adhesive) or with amalgam, but a repaired restoration is less strong than a restoration made in one piece. PMID- 11383279 TI - [Prevalence of approximal enamel lesions on bitewing radiographs in cohorts, ages 14 to 23 years]. AB - Bitewing radiographs are mainly used to detect carious lesions in the approximal tooth surfaces, and are an indispensable aid to clinical diagnosis. The aim of this study was to assess the prevalence of approximal enamel lesions from radiographs in groups of 14-, 17-, 20- and 23-years-old persons, and to determine the relation with dentine lesions and restorations present. Per age group the bitewings of 120 persons were randomly selected and assessed for enamel lesions from the distal surface of the first premolar to the mesial surface of the second molar. A second examiner assessed for enamel lesions 20% of the material to measure the interexaminer agreement (Cohen's kappa = 0.63). An enamel lesion was found in about 12 to 15% of the unfilled surfaces. The mean number of enamel lesions was almost three per person. For all age groups it was found that about 20% of the persons had at least 4 enamel lesions and the number of enamel lesions was significantly correlated with the number of dentine lesions. Of all enamel lesions 20% was found in persons without any dentinal lesions or restorations. The bitewing radiographs showed a considerable amount of enamel lesions for these age groups. This seems to justify its use on a routine base as a diagnostic tool at the age of 14-16 years. PMID- 11383280 TI - [Job stress and health in dentists]. AB - This study aimed at monitoring stressful work factors as indicated by Dutch general dental practitioners. Special attention was given to gender differences. Apart from mental pressure also physical pressure was studied. It appeared that among all dentists patient related aspects were considered stressful, als well as external interference by government, insurance company, etc. Male dentists also felt pressure from a lack of career perspective and dental office management, whereas female dentists felt pressure from unexpected or uncertain professional situations. Dentists appeared to have a good physical health, when compared with Dutch reference scores, although some job specific physical complaints could be detected. In the discussion the importance of early career planning is emphasized. PMID- 11383281 TI - [The dentist stressful for the child or the child stressful for the dentist?]. PMID- 11383282 TI - [Preoperative prosthetic treatment planning for oncology patients. Beware of pitfalls in fitting of implants!]. PMID- 11383283 TI - [Blueprint 2000 Education of dentists in the Netherlands. Objectives of undergraduate dental education. A summary]. AB - Blueprint 2000 Education of dentists in the Netherlands includes nationally developed objectives for the dentist at graduation. The changing state of oral health of the population and oral health care require a dentist trained in a broad range of dental subjects. Intellectual and scientific training should be an important aspect of undergraduate education. Developments in teaching methods support these requirements. The differences in the implementation of educational developments in the three Dutch dental schools made a uniform description of objectives not yet possible. The actual objectives therefore are presented in four different ways: (1) the profile of the dentist in broad terms, (2) general objectives by way of competencies, (3) discipline-based objectives and (4) a list of problems. These problems are derived from every day practice and can serve as starting point in the learning process of the student. It is expected, that Blueprint 2000 Education of dentists in the Netherlands will be used as a basis for curriculum development. What type of objectives, or combinations, will be used will be seen in the future. PMID- 11383284 TI - [Shooting pains in the temporal region due to infection with herpes zoster]. PMID- 11383285 TI - [BSE and other prion diseases]. PMID- 11383286 TI - [Is in a deformed jaw lowering of the 'bite' acceptable?]. PMID- 11383287 TI - Treatment options in full thickness rotator cuff tears. AB - Increasing interest in shoulder pathology during the last decades has considerably diversified the possible treatment options of full thickness rotator cuff tears. This review of the recent literature combined with information gathered during recent European shoulder meetings attempts to summarize present trends. Every full thickness cuff tear, except for the acute traumatic tear in younger patients, should always benefit first from a conservative rehabilitation program. In case of failure of the latter, reparable tears should be repaired, except perhaps in a low-demand population. This latter group of patients, presenting with a massive, irreparable tear may be satisfied with an arthroscopic debridement, but interposition techniques provide better results in activities of daily living and also give better strength. Salvage procedures such as muscle tendon transfers are technically demanding and must be reserved for the younger age group. For arthroplasty, the choice between a nonconstrained total prosthesis and a hemiprosthesis can be difficult; the early functional results of the reversed prosthesis seem to be very promising in an elderly but still active population. PMID- 11383288 TI - The bone block procedure in recurrent posterior shoulder instability. AB - We analyzed the results of eleven shoulders in ten patients who had a bone block procedure for recurrent posterior shoulder instability after extensive conservative treatment had failed. Five patients had posterior instability with additional laxity in another direction (UPI+), and five patients had unidirectional posterior instability without additional laxity (UPI-). After a median follow-up of 72 (43-102) months there was no recurrent posterior instability in the UPI-group (100% success), but the recurrence rate in the UPI+ group was high (20% success). We concluded from these results that a bone block procedure is not sufficient to treat recurrent posterior shoulder instability in unidirectional posterior instability with additional laxity but it seems to be a good method to treat unidirectional posterior instability without additional laxity. PMID- 11383289 TI - Fractures and nonunions of the carpal scaphoid in children. AB - Fractures of the carpal scaphoid in children show some quite remarkable differences with respect to fractures of the carpal scaphoid in adults. A review of the literature shows that fractures in children are more often located in the distal third, are more often incomplete and are usually not displaced. Our experience with 23 fresh fractures of the carpal scaphoid in children confirms these findings of the literature. Nonunion of the carpal scaphoid is exceedingly rare in children. When reviewing the literature we could find only 29 published cases of nonunion of the carpal scaphoid in children. In most articles describing carpal scaphoid nonunion in children, bone grafting is recommended as the treatment of choice. We report two children with a nonunion of the carpal scaphoid treated successfully by cast immobilization. Therefore, we propose that a child with a nonunion of the carpal scaphoid bone that has never been immobilized previously should be treated by cast immobilization. Surgery should be considered only if there is no indication of healing after 3 months of immobilization. PMID- 11383290 TI - Shelf acetabuloplasty for Perthes' disease: 12-year follow-up. AB - The goal of all therapies for Perthes' disease is to achieve an optimal shape of the acetabulum and an optimal coverage of the femoral head. Thirty patients who were included in this follow-up study (mean follow-up 12 years) underwent a shelf acetabuloplasty for Catterall group III or IV. The mean IOWA hip score at follow up was 96 (max. 100 points, range 74-100). The mean acetabular-head quotient increased from 82.9% pre-operatively to 107.9% postoperatively, and remained 102.4% at follow-up. There was a decrease in mean lateral subluxation ratio from 1.44 pre-operatively to 1.27 postoperatively, which remained 1.23 at follow-up. According to the Stulberg classification in the 18 adult hips, 6 hips had a good result (Stulberg 1 or 2), 10 hips had a fair result (Stulberg 3), and 2 hips had a poor result (Stulberg 4 or 5). These results appear to be better than the natural history as described by Stulberg. Shelf acetabuloplasty can be considered as an appropriate surgical treatment for severe cases of Perthes' disease. PMID- 11383291 TI - Unicondylar femoral fractures: therapeutic strategy and long-term results. A review of 23 patients. AB - Unicondylar femoral fractures account for 0.65% of all femoral fractures; they have not been studied extensively in the orthopedic literature. Usually occurring following sports trauma or traffic accidents, these fractures involve the lateral condyle three times more frequently than the medial condyle. Conservative or surgical treatment has been advocated, depending on the fracture type. From 1986 to 1999, 19 patients with unicondylar femoral fractures were surgically treated at our institution: there were 15 males and 4 females, aged 36.2 years on average. According to the AO-ASIF classification, there were 7 B1, 6 B2 and 6 B3 fractures. We used Herbert screws in six cases, Barr screws together with cancellous screws in two, cannulated screws in five, cancellous screws alone in four, a compression screw and plate in one and a T-plate in one. In the same period of time, four patients were treated conservatively with a cast. Sixteen patients treated surgically were evaluated with a mean follow-up of 60 months, using Shatzker and Lambert's criteria: six results were rated as excellent, five good, four fair and one poor, while conservative treatment gave three fair and one poor results. In conclusion, we think that open reduction and internal fixation are essential in the treatment of such fractures. PMID- 11383292 TI - [Weil's cervicocapital osteotomy for median metatarsalgia. Report of 70 cases]. AB - The authors report a series of Weil's cervicocapital metatarsal osteotomies which were performed to treat central metatarsalgias. The series included 70 central metatarsalgias treated by osteotomy of one to four metatarsals. There was an excess of length of one or several metatarsals in all cases; there were 30 metatarsophalangeal dislocations. The results were evaluated according to Kitaoka's criteria: 20 were quoted very good, 26 good, 9 fair and 3 poor. The osteotomy gave an overall improvement regarding pain and shoe fitting but the mobility of the MP joint was reduced in all cases. The results were satisfactory in cavus feet and in cases where the osteotomy was combined with correction of a hallux valgus. Weil's osteotomy allows accurate adjustment of the shortening and early weight-bearing. This is indicated in cases with excessive length of the central metatarsals and also in cases with metatarsophalangeal dislocations. PMID- 11383293 TI - Comparative study between Wilson and Mitchell metatarsal osteotomies for the treatment of hallux valgus in adults. AB - Although Mitchell osteotomy and Wilson osteotomy are two popular methods for the treatment of hallux valgus, there are no studies directly comparing their results. Fifty-six patients (73 feet) who underwent a Wilson osteotomy and 30 patients (34 feet) who had a Mitchell osteotomy were followed for a mean period of 33.7 and 38.9 months, respectively. The results were comparable in terms of hallux valgus angle correction and first intermetatarsal angle correction, although symptomatic improvement was higher in the Mitchell group. Moreover, the incidence of postoperative metatarsalgia was significantly lower in the Mitchell group as compared to the Wilson group (11.8% vs 32.9%), while less time was required for the patients who underwent Mitchell osteotomy to return to work or normal activities postoperatively. The difference in symptomatic improvement, incidence of postoperative metatarsalgia and rehabilitation time was even more clearly in favor of the Mitchell group in patients over 55. The increased stability at the osteotomy site offered by the Mitchell osteotomy compared to Wilson osteotomy could be the reason why patients had a lower incidence of postoperative metatarsalgia and returned to their normal activities faster, thus giving a higher satisfaction rate. PMID- 11383294 TI - Pullout strength of pedicle screws versus pedicle and laminar hooks in the thoracic spine. AB - While the biomechanical properties of pedicle screws have proven to be superior in the lumbar spine, little is known concerning pullout strength of pedicle screws in comparison to hooks in the thoracic spine. In vitro biomechanical pullout testing was performed to evaluate the axial pullout strength of pedicle screws versus pedicle and laminar hooks in the thoracic spine with regard to surgical correction techniques in scoliosis. Nine human cadaveric thoracic spines were harvested and disarticulated. To simulate a typical posterior segmental scoliosis instrumentation, standard pedicle hooks were used between T4 and T8 and supralaminar hooks between T9 and T12 and tested against pedicle screws. The pedicle screws were loaded strictly longitudinal to their axis; the hooks were loaded perpendicular to the intended rod direction. In total, 90 pullout tests were performed. Average pullout strength of the pedicle screws was significantly higher than in the hook group (T4-T8: 531 N versus 321 N, T9-T12: 807 N versus 600 N, p < 0.05). Both screw diameter and the bone mineral density (BMD) had significant influence on the pullout strength in the screw group. For scoliosis correction, pedicle screws might be beneficial, especially for rigid thoracic curves, since they are significantly more resistant to axial pullout than both pedicle and laminar hooks. PMID- 11383296 TI - Neurofibromatosis, gigantism, elephantiasis neuromatosa and recurrent massive subperiosteal hematoma: a new case report and review of 7 case reports from the literature. AB - The authors report the case of a 13-year-old patient with neurofibromatosis (NF I), who suffered blunt trauma to the left tibia in 1993. The diagnosis of subperiosteal hematoma was made. Treatment consisted of temporary rest. There was a recurrence in 1996, and the subperiosteal hematoma was drained. In 1997, a shortening osteotomy of the left tibia was performed. However, massive gigantism with elephantiasis of the left leg remained, causing a serious functional and cosmetic problem. In 1999, the leg was amputated above the knee. The literature is reviewed and 7 case reports are compared. The pathogenesis of subperiosteal hematoma is discussed. PMID- 11383295 TI - Surgical treatment for rheumatoid neck arthritis bilateral occipitospinal fusion with plate fixation. AB - The results of surgical treatment of 12 patients with rheumatoid cervical spine arthritis were reviewed. The Ranawat classification was as follows: 5 with Ranawat IIIB, 1 Ranawat IIIA and 6 Ranawat II. Decompression and fusion using autogenous iliac bone graft and double occipitospinal plate fixation was carried out on 11 of these patients; the remaining patient underwent upper cervical spine fusion using screw and wire fixation. The main indication for surgery was neurological deterioration. In three cases previous surgery had been carried out on the cervical spine. The results were assessed at a mean follow-up of 26.1 months. According to Frankel's grading the neurological recovery in patients with neurological compression was one grade. There was clinical and radiological evidence of fusion in all these patients. The following complications required further surgery: acute postoperative epidural hematoma (one patient) screw loosening (one patient) CONCLUSION: Fusion of the occiput and lateral mass of the involved cervical spine using a plate on each side provides a relatively stable fixation in patients with rheumatoid arthritis of the cervical spine. Laminectomy and adequate decompression of the neural elements can be carried out without compromising spinal stability. There is a relatively high complication rate associated with surgery for rheumatoid neck and the patient needs to be informed. PMID- 11383297 TI - Subtalar arthritis as a presenting symptom of Familial Mediterranean fever: case report and literature review. AB - Familial Mediterranean fever (FMF) is an autosomal, recessive disease affecting mainly people of Mediterranean origin. The primary pattern of FMF is acute, self resolving periodic attacks of high-grade fever, accompanied by either peritonitis, pleuritis, or arthritis and sometimes typical ankle rash that simulates erysipelas. Rare manifestations, such as pericarditis or massive knee effusion, have been reported in the literature as a presenting symptom of FMF. The final diagnosis has recently become more accurate by identification of the gene for FMF. We describe a unique presenting symptom of subtalar arthritis with no former personal or family history of FMF. A genetic evaluation revealed a 694/726 genetic variant that confirmed the diagnosis of FMF. Treatment with daily colchicine, 1 mg/day, resulted in complete resolution of all complaints. PMID- 11383298 TI - Total cleidectomy for a solitary metastasis of the clavicle. AB - The clavicle is a fairly common site of metastases of renal cell carcinoma. We report the cases of two patients with undiagnosed renal cell carcinoma who were first seen for shoulder pain secondary to a solitary clavicular metastasis. Wide resection was performed in both cases. Functional and cosmetic results were good, with no shoulder pain or neurovascular deficits. We suggest wide surgical resection of a solitary bony metastasis from renal cell carcinoma, associated with appropriate systemic treatment, because the survival may be increased. PMID- 11383299 TI - Dieterich's disease: avascular necrosis of the metacarpal head: a case report. AB - The authors report a case of non-traumatic avascular necrosis of the third metacarpal head. This very rare condition is known as Dieterich's disease and has been associated with trauma, systemic lupus erythematosus and steroid use. PMID- 11383300 TI - Scapho-capitate fracture syndrome. A case report. AB - A rare injury of the wrist, scapho-capitate fracture syndrome, in a young patient is reported. Despite early recognition of the injury and surgical intervention, the scaphoid fracture did not unite and another attempt to achieve union with bone grafting and internal fixation also failed. The wrist continued to be painful and stiff. Radiographs of the wrist, 18 months after the injury, showed nonunion of the scaphoid, avascular necrosis of the scaphoid and the lunate and carpal collapse with midcarpal joint arthritis. Due to persistent and disabling symptoms arthrodesis of the wrist had to be carried out. Possible causes for the bad outcome after this injury are discussed. We recommend open reduction for the fracture of the capitate and open reduction and internal fixation with primary bone grafting for a displaced comminuted scaphoid fracture. PMID- 11383301 TI - Delayed presentation of compartment syndrome following gastrocnemius tear. AB - The authors describe a case of acute compartment syndrome occurring twenty days following a tear of gastrocnemius. To their knowledge, this is the only such case reported where the onset of compartment syndrome was so long since of index injury. PMID- 11383302 TI - Fat and bone marrow embolism in total hip arthroplasty. AB - A number of cardiovascular and pulmonary complications have been reported to occur occasionally after insertion of a total hip prosthesis. Of the proposed causes of these reactions, the possibility of fat embolism has received considerable support. The increase in intramedullary pressure, produced by the mechanical compression of the femoral canal during the insertion of the stem, seems to be the decisive pathogenic factor for the development of emboli. Surgeons' proclivity to deny the clinical relevance of intraoperative emboli is directly related to their awareness of and their attempt to avoid this phenomenon. Depending on the preoperative clinical condition of the patient, the cardiorespiratory impairment may be subclinical for those with good reserve, or clinical for those with poor reserve. In cases with preexisting cardiorespiratory diseases, severe embolism can also lead to death. Moreover, tissue thromboplastin from bone marrow forced into the draining veins of the proximal femur during insertion of the stem leads to activation of the clotting cascade, lesions of the venous endothelium, and thrombogenesis. A correlation was found between the embolic events observed using transesophageal echocardiography and the cardiopulmonary function of the patients during the perioperative period. A modified surgical technique was designed to reduce the intramedullary pressure during insertion of the stem to prevent intraoperative embolic events. Surgical prevention of fat and bone marrow embolism can also reduce the incidence of postoperative deep vein thrombosis. PMID- 11383303 TI - A chain of editorial reflections. PMID- 11383304 TI - The prevalence of reading and spelling difficulties among inmates of institutions for compulsory care of juvenile delinquents. AB - Recent studies have focused on reading and writing disabilities among inmates in prisons and at juvenile institutions. Some studies in Sweden have demonstrated that more than half of the delinquents have serious reading difficulties, and for immigrants the situation is even worse. However, these studies have focused on small groups. Furthermore, little attention has been paid to different types of reading and writing difficulties. The main purpose of this investigation was to estimate the prevalence of reading and writing disabilities in juvenile institutions. The study analyses gender differences and differences between immigrants and Swedish pupils. The study included 163 pupils from 22 institutions and used three tests of literacy skills: word identification, spelling and reading comprehension. More than 70% showed some problems in reading and spelling. However, only 11% had serious difficulties. Moreover, the results showed that comprehension ability among immigrant boys was lower than among Swedish boys, despite the same level of word reading skill. The high prevalence of reading and writing disabilities seems primarily to be related to social and cultural factors, home backgrounds, limited school attendance and poor self esteem rather than to constitutional problems of a dyslexic nature. The implication of this conclusion may be important for the intervention process. PMID- 11383305 TI - An examination of the relationship between dyslexia and offending in young people and the implications for the training system. AB - A screening study was undertaken which involved 50 young offenders, serving sentences of various lengths, all from the largest young offenders' institution in Scotland. All 50 were screened for dyslexia and a number received a more detailed follow-up assessment. The results of the screening showed that 25 of the young offenders (50%) were dyslexic to some degree. This finding has implications for professionals, particularly in respect of follow-up assessment and support, and for politicians in relation to issues such as school experience, prison education and staff training. These issues are discussed here in relation to the background and results of the study. PMID- 11383306 TI - From inside prison. PMID- 11383307 TI - Measurement of radiated electromagnetic field levels before and after a changeover to energy-efficient lighting. AB - An energy-efficient lighting retrofit at the Food and Drug Administration (FDA) Winchester Engineering and Analytical Center (WEAC) presented the opportunity to measure the electromagnetic (EM) environments in several rooms before and after changing the fluorescent lighting systems and to compare the changes in EM fields with the proposed standard EM immunity levels. Three rooms, representing the types of work areas in the laboratory, were selected and measured before and after the lighting changeover. Electric and magnetic field measurements were taken in the extremely low frequency (ELF), very low frequency (VLF), and radio frequency (RF) ranges of the EM spectrum. In 2 rooms, ELF electric fields were reduced and VLF and RF electric fields were increased as a result of the changeover to high-frequency fixtures. A third room received low-frequency, energy-efficient fixtures during this changeover, and this change resulted in only a slight increase of the ELF electric fields. The ELF magnetic fields were greatly reduced in 2 but only slightly reduced in the third room. No significant change was seen in VLF or RF magnetic fields for any of these rooms. Some field strength measurements exceeded the proposed immunity levels recommended in the draft International Electrotechnical Commission standard IEC 60601-1-2 (rev. 2). The data show that increasing the separation distance from the fluorescent light fixtures greatly reduces the field-strength levels, limiting the potential for EM interference. PMID- 11383308 TI - Mechanical remodeling of small-intestine submucosa small-diameter vascular grafts -a preliminary report. AB - Small-intestine submucosa (SIS) is cell-free, 100-mu-thick collagen derived from the small intestine. It has been used as a vascular graft and has the highly desirable ability to be remodeled to become histologically indistinguishable from native adjacent artery. To date there has been limited reporting of its preimplantation and explant mechanical properties as a vascular graft. In this study, compliance, elastic modulus, and burst pressure were measured on preimplant-tested 5- and 8-mm SIS grafts and two 60-day remodeled grafts. Seven prefabricated grafts were implanted in the carotid (n = 7) in dogs, which were sacrificed after 55-63 days. The animals (n = 4) weighed from 22 to 27 kg. One dog received a unilateral carotid graft, and 3 dogs received bilateral carotid grafts. The fabrication technique employed hand-suturing with either nonresorbable or resorbable sutures. None of the grafts had a patency failure. Angiograms taken at 1 month and just before explantation showed uniform flow and no dilation. At the time of explantation, all carotid grafts were found to be encased in fibrous tissue. The grafts made with nonresorbable sutures showed thicker tissue growth at the suture line compared with those made with the resorbable sutures. Along the suture line, the grafts made with resorbable sutures exhibited a more natural color than those sutured with nonresorbable sutures. When the explanted carotid grafts were slit open, the lumen was white, shiny, and glistening. The grafts sutured with nonresorbable sutures exhibited small areas of fibrin and red blood cells when the suture was within the lumen. The resorbable-sutured grafts did not exhibit this response. The mean compliance (percent diameter increase for a pressure rise from 80 to 120 mm Hg) was on average 4.6% (range, 2.9%-8.6%) for the 5-mm preimplant-tested grafts. For the 8 mm preimplant-tested grafts, the increase in diameter for the same pressure rise was 8.7%, on average (range, 7.2% to 9.5%). For comparison, the small-diameter SIS graft at the time of implantation was about one half as compliant as the adjacent dog carotid artery, about 4 times more compliant than a typical vein graft, and more than 10 times more compliant than synthetic vascular grafts. The compliance measured on two 60-day carotid grafts was 10.5% and 7.2%, respectively. This is midway between the original compliance value and the compliance of a typical canine carotid artery (14%), indicating that mechanical remodeling occurred. The modulus of elasticity (E) increased exponentially with increasing pressure according to E = E0e alpha P, where E0 is the zero-pressure modulus and alpha is the exponent that describes the rate of increase in E with pressure; the unit of measure for variables E, E0, and P is g/cm2. The mean value for E0 was 4106 gm/cm2 (range, 1348-5601). The mean value for alpha was 0.0059 (range, 0.0028-0.0125). At 100 mm Hg, the mean value for E was 8.03 x 10(6) dynes/cm2 (range, 4.95-15.7 x 10(6)). For a 60-day SIS graft implant, the elastic modulus at 100 mm Hg decreased from a high value at implant time to twice that of a typical native canine carotid artery. The mean burst pressure for 5.5-mm grafts was 3517 mm Hg (range, 2069-4654). The burst pressure of the remodeled carotid grafts averaged 5660 mm Hg. The burst pressure for a typical carotid artery is about 5000 mm Hg. The results of this preliminary study complement those of previous SIS-vascular-graft studies and add a new factor, namely that the mechanical properties of the remodeled graft approach those of the vessel it replaces. PMID- 11383309 TI - When HIPAA finally comes, will clinical engineering be ready? AB - We live in interesting times. Recognize that HIPAA will impact nearly everyone in every health care organization and fundamentally change the health care industry. Implementation of HIPAA standards as well as ongoing developments in telemedicine, e-health and the Internet will continue to have a synergistic effect and further transform our industry. Biomedical technology will evolve and combine into ever-larger integrated networks. These networks will extend not only throughout the hospitals but also into the clinics, doctors' offices, and even patients' homes and workplaces. As clinical engineers, we need to stay ahead of the curve, anticipate and be prepared for these developments. We need to understand HIPAA and the implications it has for the technology we manage. We must ensure that the organizations we represent are prepared for what will undoubtedly be an exciting future. PMID- 11383310 TI - Listservs emerge as useful tool to link and educate biomedical community. PMID- 11383311 TI - The fundamentals of Ethernet communication. PMID- 11383312 TI - The golden age of clinical engineering: digital convergence and IT partnerships. PMID- 11383313 TI - Therapeutic strategies in systemic necrotizing vasculitides. PMID- 11383314 TI - Hormones after menopause? AB - The average life span of a woman is increasing and the age of menopausal onset has not changed much. The length of time that a woman spends in the postmenopausal state is thus increasing. This potential estrogen-deficient state may have certain physiologic and metabolic consequences. The onset of menopause is an excellent time to assess a women's overall health and to evaluate the benefit/risk equation of hormone replacement therapy (HRT). PMID- 11383315 TI - Bronchoscopic cryotherapy: preliminary experience. AB - Our preliminary experience with the bronchoscopic application of cryotherapy using rapid decompression of liquid nitrous oxide as cooling agent is reported. Seventeen applications through rigid bronchoscopy in twelve patients were performed. A single cryotherapy session was successful in the debulking of obstructive malignant lesions of the central airways in five patients (four non small cell carcinoma, one renal cell cancer metastasis), and in the treatment of a capillary haemangioma (one patient). Two sessions were successful in the treatment of a metastatic melanoma (one patient) and benign granulation tissue (one patient). Cryotherapy was also successful in the treatment of early bronchial cancer (carcinoma in situ) in four patients, requiring repetitive sessions in two. There were no complications or side-effects. These preliminary findings confirm the safety and efficacy of bronchoscopic cryotherapy in a variety of airway lesions. PMID- 11383316 TI - [Is there a relationship between hyperemesis gravidarum and hyperthyroidism?]. AB - This retrospective study evaluates the incidence and degree of thyroidal stimulation in patients with hyperemesis, and the correlation between thyroid function, the hCG level and the severity of the hyperemesis gravidarum. The role of antithyroidea is discussed in patients with a gestational hyperthyroidism. The degree of thyroid stimulation on the outcome of the pregnancy was studied. At admission hCG, TSH, FT4, FT3, TSI, anti-TPO and anti-Tg were determined. The severity of the hyperemesis gravidarum was evaluated by the degree of ketonuria, % weight loss, and the electrolytes and the liver function disorders. An ultrasound to confirm the gestational age and to exclude a multiple pregnancy or a trophoblastic disease was carried out. RESULTS: In a period of 1 January '91 to 31 January '94, 48 hyperemesis gravidarumpatients were admitted at the maternity. 22.9% of the hyperemesispatients had thyroid stimulation; 4 patients had a decreased TSH and an increased FT4 (group 2) and 7 patients had a decreased TSH, an increased FT4 and FT3 (group 3). The age of the mother, the parity and the gestational age at admission are comparable, and do not differ from the patients without thyroid disorders (group 1). Parameters, determining the severity of hyperemesis gravidarum, are not significantly different in the 3 groups. Treatment with antithyroidea in the group with proven hyperthyroidism (decreased TSH, increased FT4 and FT3) does not only lead to normalisation of the thyroid tests, but also to an improvement of the symptomatology. The gestational age at delivery and the birthweight of the babies are comparable in the 3 groups. CONCLUSION: In patients admitted with hyperemesis gravidarum, we found 23% of the women to present a thyroid stimulation. This hyperthyroidism differs from the auto-immune hyperthyroidism, i.e. the patients have no thyroid antibodies, no classic clinical signs of thyrothoxicosis. Probably, the absolute hCG concentration and its biological activity plays a crucial role in the thyroid stimulation. This degree of thyroid stimulation has no influence on the severity of the hyperemesis gravidarum, neither on the outcome of the pregnancy. PMID- 11383317 TI - Minor alterations in thyroid-function tests associated with diabetes mellitus and obesity in outpatients without known thyroid illness. AB - Thyroid function tests might be affected by diabetes and obesity. To evaluate the influence of these parameters in routine conditions, 72 diabetic and 53 non diabetic outpatients without known thyroid diseases or severe chronic illness were recruited over a 7-month period. For each patient, dosages of thyrotropin (TSH), total and free thyroxine (TT4 and FT4, respectively), total and free triiodothyronine (TT3 and FT3) and T3 resin uptake (T3RU) were performed by radioimmunoassays. The simultaneous influence of various parameters known to affect thyroid-function tests was evaluated by multivariate linear regression. The studied variables included gender, age, glucosteroids, estrogens, tobacco habits, iodine contacts, body mass index (BMI) and diabetes mellitus. Tobacco habits and iodine contacts did not influence any tests. As expected, estrogens induced an increase in TT4 and TT3 values (p < 0.001 and 0.020, respectively) associated with a decrease in T3RU (p < 0.001). Consequently, females had lower T3RU than males (p < 0.0001). Corticotherapy was associated with decreased TSH values (p = 0.022). TT3 and FT3 decreased with age (p < 0.001), whereas T3RU and FT4 increased (p = 0.020 and 0.004, respectively). In contrast to an increase in TSH (p = 0.006), TT4 and FT4 decreased at higher BMI levels (p = 0.018 and 0.004, respectively), which is consistent with subclinical hypothyroidism. In diabetic patients, TSH was lower than in nondiabetic subjects (p = 0.039). Thus, the present study indicates that besides known parameters such as age and drugs, thyroid-function tests can also be altered by diabetes mellitus and obesity. PMID- 11383318 TI - [Nutritional status of patients with type 1 and 2 diabetes in Belgium]. AB - The aim of this study was to evaluate the daily caloric intake as well as the distribution of carbohydrates, fat, protein and alcohol in a Belgian population with type 1 and 2 diabetes, in the view of the recent ADA recommendations. Dietary intake and repartition of nutriments was based upon the three days evaluation recall. Our results show excessive caloric intake in type 2 diabetic patients when compared with type 1 subjects (2652 +/- 850 vs. 2271 +/- 546 Kcal/day; P < 0.005). We also show an inadequate repartition of carbohydrates (41 +/- 5 and 39 +/- 6% in type 1 and 2 diabetic patients, respectively) and lipids (39 +/- 5 and 39 +/- 7%). Protein intake was 16% of the total caloric amount. Fibers intake was adequate. High consumption of alcohol was assessed in both type 1 and 2 diabetic patients. In conclusion, our data evidenced several pitfalls in the diet of type 1 and 2 diabetic patients. They emphasize the need for more (and repeated) practical diabetic information in order to improve daily nutritional habits. PMID- 11383319 TI - Evidence-based treatment of hypertension in patients with type 2 diabetes. PMID- 11383320 TI - Treatment of heterozygous familial hypercholesterolemia: atorvastatin vs simvastatin. AB - BACKGROUND AND AIM: This study compares the cholesterol-lowering efficacy of atorvastatin and simvastatin in attainment of the National Cholesterol Education Program (NCEP) guidelines LDL-cholesterol (LDL-C) goal in patients with heterozygous familial hypercholesterolemia (HFH). The association of atorvastatin with significant changes of blood fibrinogen and other coagulative variables was also compared with that of simvastatin. METHODS AND RESULTS: In a 24-week study, 26 HFH patients (16 men, 10 women, mean age 55.1 +/- 11.3) were randomly assigned to receive atorvastatin or simvastatin. The initial daily dose of 10 mg was progressively raised to 20, 40 and 80 mg in patients who had not reached the NCEP LDL-C goal. Significant reductions of total and LDL-C (p < 0.001), triglycerides (p < 0.005) and apoB100 (p < 0.001) were observed in both groups. Atorvastatin caused greater reductions in total cholesterol (-42% vs -30%) (p < 0.001) and LDL C (-50% vs -37%) (p < 0.01). Three patients treated with Atorvastatin (23%) and none of those treated with simvastatin reached the NCEP LDL-C goal at the end of the study. No significant departures from the fibrinogen and coagulative variable baselines were observed. CONCLUSIONS: Atorvastatin has greater cholesterol lowering efficacy than simvastatin in HFH. PMID- 11383322 TI - Gene-environment interactions: the example of coagulation factors. PMID- 11383321 TI - Effects of de-alcoholated red wine and its phenolic fractions on platelet aggregation. AB - BACKGROUND AND AIM: Platelet aggregation is involved in atherosclerosis and pharmacological inhibition of platelet activity may reduce the risk of coronary thrombosis and myocardial infarction. Red wine polyphenols may reduce platelet aggregability. This study evaluates the effect of de-alcoholated red wine (DRW) and its phenolic fractions on rat platelet aggregation and cyclic AMP (c-AMP) content. METHODS AND RESULTS: DRW was fractionated into four classes of phenolic compounds: phenolic acids (fraction 1), procyanidins, catechins and monomeric anthocyanidins (fraction 2), flavonols and resveratrol (fraction 3) and polymeric anthocyanidins (fraction 4). The effect of each fraction on ADP-induced rat platelet aggregation and c-AMP content was compared with that of DRW and pure phenolic compounds (quercetin, catechin, resveratrol, caffeic acid). DRW completely inhibited ADP-induced platelet aggregation. Fraction 2 also showed a significant anti-aggregating activity, whereas the effects of fractions 3 and 4 and the pure phenolics were not significant. A significant increase in platelet c AMP content was observed after the addition of DRW and fraction 2. CONCLUSIONS: Our data indicate that DRW and its catechin-anthocyanidin fraction exert a significant effect on platelet aggregation in vitro, perhaps by enhancing platelet c-AMP levels. PMID- 11383323 TI - Effects of plant sterols and stanols on lipid metabolism and cardiovascular risk. AB - Functional foods enriched with plant sterols and stanols are on sale in many countries. Due to their structural similarity with cholesterol, these additives lower intestinal absorption of cholesterol, resulting in a 10-15% reduction in LDL-cholesterol when their daily intakes are 2-3 g. They are also effective as part of a cholesterol-lowering diet and in combination with cholesterol-lowering drugs. Estimates for the absorption of plant sterols (sitosterol and campesterol) and of campestanol are around 10%, and for sitostanol less than 5%. Lipid standardized plasma levels are very low, but increase when statins are used. Extensive toxicological evaluation studies have not revealed any harmful side effects. In human studies, side-effects were comparable to placebo treatment. However, lipid-standardized levels of the hydrocarbon carotenoids may decrease, without leaving the normal range. Together, these findings indicate that these functional foods have great potential in the prevention of coronary heart disease. However, post-marketing surveillance for example for functional foods in general is necessary to monitor possible adverse effects and describe consumers and consumption patterns. PMID- 11383324 TI - Antithrombotic drugs for older subjects. Guidelines formulated jointly by the Italian Societies of Haemostasis and Thrombosis (SISET) and of Gerontology and Geriatrics (SIGG). AB - Older individuals contribute heavily to the percentage of deaths due to myocardial infarction (MI) and stroke. The incidence of venous thromboembolism (VTE) is highest in subjects > 65 years. Prospective intervention trials involving groups of clinically comparable subjects > or = 60 allow the following statements to be made with regard to the use of antithrombotic drugs in the elderly. Antiplatelet agents. To prevent recurrence of ischaemic stroke and MI in stable/unstable angina, MI, TIA/stroke or peripheral arterial disease, aspirin is the drug of choice. Clopidogrel is more effective than aspirin in this respect. Heparin. For the treatment of acute deep venous thrombosis (DVT) and pulmonary embolism (PE), intravenous standard heparin or subcutaneous standard heparin are effective (aPTT 1.5-2.0 times baseline values). As the risk of bleeding increases with age, low-molecular-weight heparins (LMWH) are preferable in the elderly. For the prophylaxis of VTE in general surgery in subjects at low-moderate risk, low dose heparin or low doses of LMWH are effective. In subjects at high risk, adjusted-dose heparin plus physical devices or high-dose LMWH are recommended. The combination of heparin and aspirin is the standard treatment for unstable angina and non-Q wave MI. LMWH are as active as standard heparin in this indication. Vitamin K antagonists. For the chronic treatment of VTE, warfarin is also the treatment of choice (INR 2.0-3.0) in the elderly, though lower doses are needed due to their hypersensitivity to oral anticoagulants. For the prevention of thromboembolic stroke in patients > 75 with atrial fibrillation, warfarin is the drug of choice. Patients aged 65-75 may receive warfarin or aspirin. Thrombolytic agents. Thrombolytic agents are not recommended for treating DVT in the elderly because of their limited risk/benefit ratio and should be confined to massive PE. In the absence of contraindications, thrombolysis for MI may be considered in the elderly. PMID- 11383325 TI - PPAR gamma: an essential role in metabolic control. AB - The peroxisome proliferator-activated receptor gamma is a nuclear hormone receptor playing a crucial role in adipogenesis and insulin sensitization. Prostaglandin J2 derivatives and the antidiabetic thiazolidinediones are its respective natural and synthetic ligands. The RXR/PPAR gamma heterodimer has also been reported to have important immunomodulatory activities and its pleiotropic functions suggest wide-ranging medical implications. PMID- 11383326 TI - Effects of atorvastatin and omega-3 fatty acids on LDL subfractions and postprandial hyperlipemia in patients with combined hyperlipemia. AB - BACKGROUND AND AIM: The aim of the present study was to see whether a moderate dose of omega-3 fatty acids (FA) potentiates the beneficial effects of statins on the high risk for coronary heart disease (CHD) in patients with combined hyperlipemia. METHODS AND RESULTS: In the present double-blind parallel study, 42 patients with combined hyperlipemia with serum triglycerides 2-15 mmol/L-1 and serum total cholesterol > 5.3 mmol/L-1 at the end of a three-month dietary run-in period were treated with 10 mg/d atorvastatin for 10 or more weeks. During the last 5 weeks they were randomized into two groups that received either 1.68 g/d omega-3 FA as ethylesters of eicosapentaenoic (45%) and docosahexaenoic acids (39%), or placebo (corn oil). As expected, atorvastatin significantly reduced serum total LDL-cholesterol (LDL-C), triglycerides and apolipoproteins B, E, CII and CIII, whereas HDL-cholesterol (HDL-C) was increased. Addition of omega-3 FA further increased HDL-C (p < 0.03), and reduced systolic blood pressure (< 0.03), while the small dense LDL-particles (LDL III) (p < 0.05) and postprandial hypertriglyceridemia (p < 0.01) were reduced compared with the baseline, though there were no significant differences to the placebo group. This may be related to the large individual variation in these parameters and the small number of patients. No significant effects on basic or postheparin activities of lipoprotein lipase or hepatic lipase were observed after atorvastatin with or without addition of omega-3 FA. CONCLUSIONS: This study indicates that addition of a low dose of omega-3 FA may further improve the risk profile for CHD in patients with combined hyperlipemia treated with atorvastatin. The effect is related to reduction of postprandial hyperlipemia and redistribution of LDL subfractions. PMID- 11383327 TI - Mitochondrial uncoupling proteins (UCPs) and obesity. AB - Obesity is now regarded as major public health problem worldwide. Research into this condition has been increasingly focussed on elucidating the cellular and molecular mechanisms regulating mammalian energy intake and expenditure. It is widely acknowledged that the brown adipose tissue (BAT) mitochondrial uncoupling protein (UCP1) plays a pivotal role in adaptive thermogenic responses. Two homologues of UCP1 (UCP2 and UCP3) have recently been identified and population based genetic studies have linked them with basal metabolic rate, while in vitro studies report that both have proton transport activity and may thus be involved in regulation of energy homeostasis and hence obesity. However, evidence from genetically modified animal models indicates that UCP2 and UCP3 have no specific physiological thermogenic function in vivo, though they may still be useful therapeutic targets for obesity. Furthermore, their role in modulating levels of reactive oxygen species and glucose homeostasis is also being investigated. PMID- 11383328 TI - Leaching of heavy metals from contaminated soils using EDTA. AB - Ethylenediaminetetraacetic acid (EDTA) extraction of Zn, Cd, Cu and Pb from four contaminated soils was studied using batch and column leaching experiments. In the batch experiment, the heavy metals extracted were virtually all as 1:1 metal EDTA complexes. The ratios of Zn, Cd, Cu and Pb of the extracted were similar to those in the soils, suggesting that EDTA extracted the four heavy metals with similar efficiency. In contrast, different elution patterns were obtained for Zn, Cd, Cu and Pb in the column leaching experiment using 0.01 M EDTA. Cu was either the most mobile or among the most mobile of the four heavy metals, and its peak concentration corresponded with the arrival of full strength EDTA in the leachate. The mobility of Zn and Cd was usually slightly lower than that of Cu. Pb was the least mobile, and its elution increased after the peaks of Cu and Zn. Sequential fractionations of leached and un-leached soils showed that heavy metals in various operationally defined fractions contributed to the removal by EDTA. Considerable mobilisation of Fe occurred in two of the four soils during EDTA leaching. Decreases in the Fe and Mn oxide fraction of heavy metals after EDTA leaching occurred in both soils, as well as in a third soil that showed little Fe mobilisation. The results suggest that the lability of metals in soil, the kinetics of metal desorption/dissolution and the mode of EDTA addition were the main factors controlling the behaviour of metal leaching with EDTA. PMID- 11383329 TI - Modelling the sorption of metals from aqueous solutions on goethite fixed-beds. AB - The research on separation methods of toxic metals from wastewater streams is continuous and intensive and, among them, sorption processes are considered of particular importance, mainly due to their effectiveness. The sorption of chromate anions and zinc cations from dilute aqueous solutions using a packed-bed (column configuration) of synthesised granulated goethite was investigated in the present study. The examined parameters during this investigation were the following: (1) the initial pH value of metal-laden aqueous solution (two representative values at acidic pH, 3.5 and 5.0, were tested); (2) the quantity of sorbent in the column, corresponding to bed height; and (3) the influence of ethylene diamine tetra acetic acid (EDTA) addition, a common, strong chelating agent. The Bed Depth-Service Time model has been applied to the sorption results in order to model the column operation. The removal efficiency of Cr(VI) anions by the sorptive column was found to be higher than that of Zn(II) cations. The presence of EDTA caused a certain decrease of removal efficiency for the case of hexavalent chromium, due to competition for the same sorption sites, while the removal of divalent zinc was increased, due to variation of cation speciation. PMID- 11383330 TI - PCDDs in the water/sediment-seagrass-dugong (Dugong dugon) food chain on the Great Barrier Reef (Australia). AB - Polychlorinated dibenzo-p-dioxin (PCDD) and dibenzofuran (PCDF) concentrations were measured in sediment and seagrass from five locations in or adjacent to the Great Barrier Reef Marine Park. A full spectrum of Cl5-8DDs were present in all samples and, in particular, elevated levels of Cl8DD were found. PCDFs could not be quantified in any samples. The PCDD concentrations ranged over two orders of magnitude between sites, and there was a good correlation between sediment and seagrass levels. There were large quantities of sediment present on the seagrass (20-62% on a dry wt. basis), and it was concluded that this was a primary source of the PCDDs in the seagrass samples. The PCDD levels in the seagrass samples were compared with the levels in the tissue of three dugongs stranded in the same region. The relative accumulation of the 2,3,7,8-substituted PCDD congeners in the dugongs decreased by over two orders of magnitude with increasing degree of chlorination. This was attributed to the reduced absorption of the higher chlorinated congeners in the digestive tract, a behaviour that has been observed in other mammals such as domestic cows. PMID- 11383331 TI - Isotherms and sequential extraction procedures for evaluating sorption and distribution of heavy metals in soils. AB - Heavy metals are potentially toxic to human life and the environment. Their contaminating effect in soils depends on chemical associations. Hence, determining the chemical form of a metal in soils is important to evaluate its mobility and bioavailability. We utilized a sequential extraction procedure and sorption isotherms (monometal and competitive) to evaluate the mobility and distribution of Cd, Cu, Ni, Pb, and Zn in four soils differing in their physicochemical properties: Calcixerollic Xerochrepts (Cx1 and Cx2), Paralithic Xerorthent (Px) and Lithic Haplumbrept (Lh). Most of the metals retained under point B conditions of sorption isotherms were extracted from the more mobile fractions: exchangeable and carbonates, in contrast with the profiles of the original soils where metals were preferently associated with the residual fraction. In soils having carbonate concentration under 6% (Cx1 and Lh), the exchangeable fraction was predominant, whereas in calcareous soils (Cx2 and Px) metals extracted from carbonates predominated. Partitioning profiles were in accordance with the affinity sequences deduced from the initial slope of isotherms and showed that the soils had a greater number of surface sites and higher affinity for Pb and Cu than for Cd, Ni, or Zn. In general, the simultaneous presence of the cations under study increased the percentages of metals released in the exchangeable fraction. The tendency towards less specific forms was more noticeable in Cx2 and Px soils and for Ni, Zn, and Cd. The affinity of inorganic surfaces was larger for Zn than for Cd or Ni, but the affinity of organic surfaces was larger for Cd or Ni than for Zn. PMID- 11383332 TI - Preliminary measurements of summer nitric acid and ammonia concentrations in the Lake Tahoe Basin air-shed: implications for dry deposition of atmospheric nitrogen. AB - Over the past 50 years, Lake Tahoe, an alpine lake located in the Sierra Nevada mountains on the border between California and Nevada, has seen a decline in water clarity. With significant urbanization within its borders and major urban areas 130 km upwind of the prevailing synoptic airflow, it is believed the Lake Tahoe Basin is receiving substantial nitrogen (N) input via atmospheric deposition during summer and fall. We present preliminary inferential flux estimates to both lake surface and forest canopy based on empirical measurements of ambient nitric acid (HNO3), ammonia (NH3), and ammonium nitrate (NH4NO3) concentrations, in an effort to identify the major contributors to and ranges of atmospheric dry N deposition to the Lake Tahoe Basin. Total flux from dry deposition ranges from 1.2 to 8.6 kg N ha-1 for the summer and fall dry season and is significantly higher than wet deposition, which ranges from 1.7 to 2.9 kg N ha-1 year-1. These preliminary results suggest that dry deposition of HNO3 is the major source of atmospheric N deposition for the Lake Tahoe Basin, and that overall N deposition is similar in magnitude to deposition reported for sites exposed to moderate N pollution in the southern California mountains. PMID- 11383333 TI - Monitoring laboratory-scale bioventing using synchronous scan fluorescence spectroscopy: analysis of the vapor phase. AB - Bioventing is an improved method of soil remediation that is being used with increasing frequency. In this paper, we refine techniques to measure the progress of petroleum hydrocarbon decomposition by monitoring vapor phase composition with synchronous scan fluorescence spectroscopy (SSFS). Analysis of the vapor phase has advantages compared to standard extraction techniques that require extensive sample handling and clean up. For comparison, hydrocarbon contamination in the soil was measured by analysis of Soxhlet extractions with gas chromatography-mass spectrometry (GC-MS). Comparison of the GC-MS and SSFS data showed that changes in hydrocarbon composition measured in the vapor phase provide an accurate measure of decomposition reactions taking place in the soil. PMID- 11383334 TI - Dynamics of PAH deposition, cycling and storage in a mixed-deciduous (Quercus Fraxinus) woodland ecosystem. AB - Estimates of standing biomass and fluxes of biomass in a mixed-deciduous woodland were derived, and used with results for concentrations of seven polycyclic aromatic hydrocarbons (PAHs) in different compartments of the woodland system to quantitatively assess some of the key fluxes and burdens of PAHs in this complex system. We quantified PAH burdens in air, in leaves of three deciduous tree species, in leaf litter and in soil, and uptake of PAHs by the tree leaves; PAH fluxes in litterfall, and deposition to the litter layer on the woodland floor during winter were calculated from these data. Air burdens exhibited marked seasonal variations for all compounds, with lowest values in summer when combustion-related emissions were low. Leaves did not accumulate large burdens of PAHs while on the trees and consequently, litterfall-associated fluxes of PAHs were small, representing only a fraction of the burdens in the litter layer to which they were deposited. Higher PAH burdens in air in winter, combined with the organic-matter-rich nature of the litter layer, are thought to be responsible for fluxes of PAHs to the litter layer in winter being 20-170 times the peak litterfall fluxes. The soil compartment was calculated to contain 25 years' worth of deposition of benzo[ghi]perylene, the most recalcitrant PAH in this study. Storage quotients for fluoranthene, pyrene, benzo[k]fluoranthene and benzo[a]pyrene burdens in soil represented 7-10 years' worth of deposition, while fluorene and phenanthrene storage in soil approached unity with inputs (1 and 3 years' worth of deposition, respectively). The relative importance of storage and loss processes was therefore closely related to the physico-chemical properties of the PAH, and is discussed in relation to the cycling of carbon in the woodland. PMID- 11383335 TI - Variation of gas exchange within native plant species of Switzerland and relationships with ozone injury: an open-top experiment. AB - Gas exchange and ozone-induced foliar injury were intensively measured during a 6 day period in mid-August 1998 on leaves of Acer pseudoplatanus, Betula pendula, Corylus avellana, Fagus sylvatica, Fraxinus excelsior, Morus nigra, Prunus avium, Prunus serotina, Rhamnus cathartica, and Viburnum lantana at a forest nursery site in Canton Ticino, Switzerland. Plants were grown in four open plots (AA), four open-top chambers receiving carbon-filtered (CF) air, and four receiving non filtered (NF) air. Significant variation in gas exchange (F > 12.7, P < 0.001) was detected among species with average net photosynthesis and average stomatal conductance differing by a factor of two. Species also varied significantly in foliar injury for those leaves for which we measured gas exchange (F = 39.6, P < 0.001). Fraxinus excelsior, M. nigra, P. avium, P. serotina, R. cathartica, and V. lantana showed more injury than A. pseudoplatanus, B. pendula, C. avellana, and Fagus sylvatica. Plants grown in CF chambers had significantly higher net photosynthesis (A) and stomatal conductance to water vapor (gwv), and lower foliar injury than plants grown in NF chambers and AA plots; interactions between species and ozone treatments were significant for all variables (F > or = 2.2, P < 0.05) except gwv (F = 0.7, P > 0.1). Although A and gwv decreased and foliar injury increased with leaf age, the magnitude of these changes was lower for plants grown in CF chambers than for plants grown in NF chambers and AA plots. Neither ozone uptake threshold (r = 0.26, P > 0.20) nor whole-plant injury (r = 0.15, P > 0.41) was significantly correlated with stomatal conductance across these species. It appears that the relationships between stomatal conductance and foliar injury are species-specific and interactions between physiology and environments and leaf biochemical processes must be considered in determining species sensitivity to ambient ozone exposures. PMID- 11383336 TI - Biomagnification of organochlorines along a Barents Sea food chain. AB - To trace the biomagnification of organochlorines in marine food chains near Svalbard, which may lead to the high organochlorine concentrations in top predators from the area, we compared concentrations and patterns of organochlorines in selected taxa. The pelagic crustaceans, Calanus spp. (copepods), Thysanoessa spp. (euphausiids), Parathemisto libellula (amphipod), and the fish species, Boreogadus saida (polar cod) and Gadus morhua (cod) were selected to represent the lower trophic levels in the food web. Four seabird species were chosen at the higher trophic levels, Uria lomvia (Brunnich's guillemot), Cepphus grylle (black guillemot), Rissa tridactyla (black-legged kittiwake) and Larus hyperboreus (glaucous gull). We found low concentrations of the organochlorines sigma hexachlorocyclohexanes (sigma HCHs), hexachlorobenzene (HCB), sigma Chlordanes, sigma DDTs and sigma polychlorinated biphenyls (sigma PCBs) in crustaceans (11-50 ng g-1 lipid wt.) and fish (15-222 ng g-1 lipid wt.). In seabirds, the organochlorine concentrations biomagnified one to three orders of magnitude dependent on species and compound class. Glaucous gulls had the highest concentrations of all organochlorines. The organochlorine levels in all taxa except glaucous gull were comparable to those recorded in similar species in the Canadian Arctic. The organochlorine pattern changed from crustaceans and fish to seabirds. Moving up the food chain, the relative contribution of sigma HCHs, HCB and sigma Chlordanes decreased, and the relative contribution of sigma DDTs, sigma PCBs, persistent compounds and metabolites increased. The results reflected trophic transfer of organochlorines along the food chain as well as different elimination potentials due to direct diffusion in crustaceans and fish, and higher contaminant metabolic activity in seabirds. PMID- 11383337 TI - Influence of trophic status on PCB distribution in lake sediments and biota. AB - We investigated the relationship between trophic status and polychlorinated biphenyl (PCB) distribution in 19 Swedish lakes. We analyzed PCB in water, phytoplankton, zooplankton, fish and sediment during two sampling periods, in spring and summer. The mass of sigma PCB in the lake sediments was positively related to lake trophy, i.e. more PCBs were accumulated and buried in the sediment of eutrophic lakes than in oligotrophic lakes. In the oligotrophic lakes a greater fraction of the total PCB load was dissolved in water. We conclude that this is a result of higher sedimentation rates in eutrophic lakes and relatively lower turnover of organic carbon in the water column of the shallow, eutrophic lakes. In the stratified lakes, the amount of PCB per cubic meter in the epilimnion decreased from spring to summer. We suggest that sedimentation of plankton beneath the thermocline during stratification act as a sink process of PCBs from the epilimnion. PMID- 11383338 TI - Geochemistry of natural and anthropogenic metals in the coastal sediments of the island of Lesvos, Aegean Sea. AB - The geochemistry of metals in the harbor and coastal zone of the town of Mytilene (island of Lesvos, Aegean Sea, Greece) was studied after normalization of the metal data to a conservative element. In the study area, Li was proven to be better suited than Al for such normalization and it was able to describe successfully the natural metal variability of the coastal sediments. Metal contamination (Cd, Cu, Pb, Zn) was recorded in the harbor sediments, while no pollution signs were detected in the wider coastal zone or the sediments of the ancient harbor, at the northern part of the town. The geochemical normalization of metal data to Li is a method that can detect the degree of metal contamination taking into consideration the natural metal variability in the sediments of a study area. PMID- 11383339 TI - The asexual enchytraeid worm Cognettia sphagnetorum (Oligochaeta) has increased Cu resistance in polluted soil. AB - We studied Cu resistance in the asexual (reproduction through fragmentation) enchytraeid worms (Cognettia sphagnetorum, Oligochaeta) originating from two sites: one uncontaminated, and another contaminated by heavy metals. Adult worms were smaller and population density was lower at the polluted site. However, adults from the contaminated site had better survival in Cu-contaminated soil, but lower survival as juveniles (fragments). As we do not know the genetic basis of Cu resistance of the worms, it may have been reached by acclimatization via induced Cu regulation. Because fragmentation is the only mode of reproduction, all phenotypic properties (including resistance) of a parental generation could be transferred to filial generations. PMID- 11383340 TI - Chlordane components and metabolites in seven species of Arctic seabirds from the Northwater Polynya: relationships with stable isotopes of nitrogen and enantiomeric fractions of chiral components. AB - The Northwater Polynya (NOW) is a large area of year-round open water found in the high Arctic between Ellesmere Island and Greenland. NOW has high biological productivity compared with other arctic marine areas, and supports large populations of several seabird species. Seven species of seabirds, dovekie (Alle alle, DOVE), thick-billed murre (Uria lomvia, TBMU), black guillemot (Cepphus grylle, BLGU), black-legged kittiwake (Rissa tridactyla, BLKI), ivory gull (Pagophila eburnea, IVGU), glaucous gull (Larus hyperboreus, GLGU) and northern fulmar (Fulmaris glacialis, NOFU) were collected in May and June 1998 to determine chlordane concentrations in liver and fat and to examine species differences, relationships with stable isotopes of nitrogen, and enantiomeric fractions (EFs) of chiral components. sigma CHLOR concentrations varied over an order of magnitude among species, from a low of 176 +/- 19 ng/g (lipid corrected) in TMBU liver to a high of 3190 +/- 656 ng/g (lipid corrected) in NOFU liver. Lipid-corrected concentrations of chlordane did not vary between sex for any species or between fat and liver except for the DOVE, that had fat concentrations that were significantly greater than the liver. delta 15N values described a significant percentage of the variability of concentrations for most chlordane components, although less than what has been reported for whole food chains. Slopes of delta 15N versus concentration of chlordane components and sigma CHLOR were similar with the exception of those which were metabolized (trans-chlordane) or formed through biotransformation (oxychlordane). The relative proportions of chlordane components in seabirds were related to phylogeny; the procellariid (NOFU) had the greatest percentage of oxychlordane (> 70%), followed by the larids (BLKI, IVGU and GLGU; 40-50%) and the alcids (DOVE and BLGU; 10-20%). The exception was TBMU, an alcid, where oxychlordane made up > 40% of its chlordane. EFs of chiral components failed to predict concentration or trophic level, but did identify biotransformation differences between species and chlordane components. TBMU appeared to have a greater capacity to metabolize and eliminate chlordane, based on high proportions of oxychlordane, the highest EFs for oxychlordane and heptachlor epoxide, and a delta 15N-sigma CHLOR value which was well below the relationships developed for all seabird species. PMID- 11383341 TI - Time to death response in carabid beetles exposed to multiple stressors along a gradient of heavy metal pollution. AB - We investigated the responses of invertebrates inhabiting polluted environments to multiple stressors. Carabid beetles (Pterostichus oblongopunctatus F.) were subjected to food deprivation and insecticide treatment (dimethoate) to resolve trends associated with a gradient of heavy metal pollution. Metal concentrations along the gradient of five sites ranged from approximately 150 to 10,500 mg/kg Zn, 136 to 2600 mg/kg Pb, and 0.84 to 81.9 mg/kg Cd. There was no difference in body mass along the pollution gradient. However, the beetles originating from the most contaminated sites were significantly less tolerant to food deprivation than beetles from the reference site. Median survival time was 120 h for the two most polluted sites, compared with 168 h at the reference site. Beetles from the two most polluted sites were also significantly more susceptible to dimethoate at 0.1 microgram active ingredient/beetle. Median survival times were 12 and 123 h for beetles from the two most polluted sites and 359 h for the reference site. Carabid beetles exposed to chronic pollution, therefore, exhibit elevated susceptibility to additional stressors. PMID- 11383342 TI - Patient advocacy. Winning ways. AB - Patient advocates need considerable professional experience and personal skills. They should report to someone at board level. The individual appointed should be prepared for initial wariness from other staff and have a non-judgemental approach to staff and clients. The seniority of the post should be reflected in the salary. PMID- 11383343 TI - Cross-border healthcare. Going nowhere. AB - Expectations of significant numbers of patients crossing borders within the EU for treatment have not materialised. This is despite the establishment of projects in border regions designed to increase the number of patients going outside their own country for treatment. Doctors' reluctance to refer patients outside their own country may be a factor in this. There are still legal and administrative uncertainties about reimbursement for care in another country. PMID- 11383344 TI - Day surgery. All in a day's work. PMID- 11383345 TI - Wound care. Painful subject. PMID- 11383346 TI - Guide to nicotine replacement therapies. Breath of fresh air. PMID- 11383347 TI - Rehabilitation. Upwardly mobile. PMID- 11383348 TI - [Split wisdom tooth high in the upper jaw]. PMID- 11383349 TI - [Legionellosis]. PMID- 11383350 TI - [A personal guide for medical-dental information]. PMID- 11383351 TI - [Distal support in a free-ending frame prosthesis]. PMID- 11383352 TI - [Repair and revision 2. Repair or replacement of composite]. AB - Nowadays adhesive techniques enable the dentist to preserve sound tooth tissue as only decay has to be removed when a carious lesion has to be restored. Also, when an existing composite resin restoration shows a defect, a total replacement is not always necessary and a repair can be made. Specific techniques for repairing old composite resin restorations are etching with hydrofluoridic acid, sandblasting and application of a silane solution. The decision between repair or replacement is made based on the quality of the existing restoration, the needed repair strength, the biological price of a total replacement and the skills of the operator. Some case reports are added as illustrations. PMID- 11383353 TI - [National Health Services in the Netherlands: its influence on dental care]. AB - In the year 1941 the National Health Services Act came into force in which the insured patients have claims to dentistry. The influence on dental care is beyond any doubt and is illustrated by some examples. The dental care for the youth is mentioned in particular. Special attention is paid to legal regulations concerning the application of fluorides. PMID- 11383354 TI - [Reparation and revision 3. Perforations during root canal treatment]. AB - Iatrogenic perforations from the pulp cavity towards the periodontium result in complications that often lead to root or tooth removal. Identification of perforations is possible by direct observation of bleeding, indirect bleeding assessment using paperpoints, radiography and an apex locator. Successful treatment depends mainly on immediate sealing and prevention of infection. Several factors affect the achievement of these goals, such as time of occurrence, size and location of the perforation and the lability of the repair material to seal the lesion. An overview is presented of diagnosis, prognosis and non-surgical treatment of perforations. PMID- 11383355 TI - [Three treatment modalities for mandibular overdentures on implants. Clinical and radiological aspects]. AB - The results are presented of 3 treatment modalities for overdentures on implants in the mandible: 2 implants with ball attachment, 2 implants interconnected with a bar and 4 implants interconnected with bars. The patients, in total 110, were at random treated and evaluated 3, 9 and 19 months after insertion of the implants. Six implants were lost out of 283 implants during the osseointegration period. There were hardly any differences found between the groups with respect to the clinical and radiologic parameters. De first group showed less bleeding around the implants after 19 months then the other groups. Around the medial implants there was significantly more bone resorption then around the distal implants and the implants in the other groups. No conclusions can be drawn with respect to the best treatment modality since the period of evaluation is too short and the other parameters like patients' experiences are not included. PMID- 11383356 TI - [New film for intraoral radiography]. AB - Recently, a new film for intraoral radiography was introduced, the Kodak Insight Dental film. The manufacturer claims that this film requires a reduced exposure time compared to Ektaspeed Plus-film, whereas other relevant film characteristics remained unchanged. These claims were investigated in this study. The exposure time of the Insight-film was on average 17% less than that of Ektaspeed Plus-film upon standard manual development, and 49% less than that of Kodak Ultraspeed film. Compared to the E-speed film, the Insight-film has an identical gradient and spatial resolution (> 10 lp/mm). Compared to the D-speed film the gradient was larger, indicating a better contrast, whereas the resolution was the same. It was concluded that the Insight Dental film can be used in intraoral radiographic diagnosis. PMID- 11383357 TI - [Cloning: applications in humans 1. Technical aspects]. AB - The successful cloning experiments in mammals such as the sheep and mouse prompted speculations on clinical application in humans. Cloning is possible by nucleus transplantation and by embryo splitting. Nucleus transplantation does not result in a genetically completely identical individual because the mitochondrial DNA originates from the ovum donor. Embryo splitting may be regarded as the artificial production of a monozygotic multiplet. Possible applications of cloning in humans belong in the context of reproduction (treatment of couples with subfertility, with genetic problems or with a 'replica motive'), transplantation of genetically identical tissue, and scientific research. PMID- 11383358 TI - [Swollen tongue]. PMID- 11383359 TI - [Depression]. PMID- 11383360 TI - [Advantages and disadvantages of intraosseous anesthesia]. PMID- 11383361 TI - [Supplemental fat in the diet of horses...is it advantageous?]. AB - Feeding fat-supplemented diets to horses has drawn considerable interest. One of the advantages of such diets is that the energy density is increased, so that less feed is needed to meet energy requirements. In addition, adding fat to the diet enhances the contribution of fat oxidation to energy production, thus sparing muscle glycogen. The 'spared' glycogen is available for energy metabolism when the acutely exercising horse reaches a point of oxygen deficit and must rely on anaerobic metabolism. This appears to be beneficial for both aerobic and anaerobic performance. Fats are readily digested by the horse. Vegetable oils are more palatable to horses than animal fats, but the palatability of fat-rich diets may decrease in the long term. PMID- 11383362 TI - [The sharpening and polishing of dental instruments]. AB - This article describes the maintenance of dental instruments in a companion animal practice, in particular the correct sharpening techniques for tartar scrapers, curettes, excavators, and elevators. Sharp instruments are essential for the quality and speed of dental care. PMID- 11383363 TI - [Traveling with pets. MAFF announces revision of quarantine regulations]. PMID- 11383364 TI - [Who is the bitten dog? Legal liability for damage by animals. Part 2]. PMID- 11383365 TI - [Not all veterinarians have been sleeping]. PMID- 11383367 TI - [Enormous excitement]. PMID- 11383366 TI - [Homeopathic veterinarians take advantage of foot and mouth disease crisis]. PMID- 11383368 TI - [Advertisement Palfium = Palface]. PMID- 11383369 TI - [Homeopathic medicine to control foot and mouth disease outbreak is not sustained]. PMID- 11383370 TI - [Anesthesia problems]. PMID- 11383371 TI - ["Permanent education; when are we allowed to proceed again?"]. PMID- 11383372 TI - Should all patients with cardiovascular disease receive statin therapy? AB - With the strong correlation between the development of coronary heart disease and elevated levels of total cholesterol and low-density lipoprotein cholesterol (LDL C), therapies that significantly lower lipid levels will be widely prescribed. Within the past 15 years, major studies have shown the statins to be very effective in lowering LDL-C levels. Are the effects of statin therapy powerful enough to justify administering one of these drugs to every patient with cardiovascular disease? Among the arguments favoring its general usage are its safety record, its high rate of patient compliance (especially in comparison to alternative therapies such as diet and exercise), and its cost effectiveness. On the other hand, elevated levels of LDL-C are not the only cause of atherosclerosis, so simply lowering the LDL-C level is not the sole answer to reducing the risk of mortality and morbidity from coronary heart disease. PMID- 11383373 TI - Threshold level or not for low-density lipoprotein cholesterol. AB - As drugs, such as the statins, and other therapies demonstrate the ability to significantly lower levels of low-density lipoprotein cholesterol (LDL-C), one issue is whether there is a lower threshold below which no further decline in coronary heart disease occurs. Those who evaluate the data from multiple trials and conclude that no significant decrease in coronary event rates occurs at or below 125 mg/dL suggest using this level as a guideline for clinical application of cholesterol-lowering therapy. On the other hand, analysis of the results of the same population and primary prevention studies concludes that no such threshold exists. The issues affected by the decision of whether to use a threshold include costs to the healthcare system for additional physician time, tests, and medication; unknown clinical events and safety related to very low LDL C; and resource prioritization to an unestablished therapeutic approach. PMID- 11383374 TI - Statins--similarities and differences. AB - The number of statins available to physicians continues to grow, leading to the question: Are all statins alike? Comparisons of side effects and safety profiles and the dose-response relationship among the different drugs show similar results. The cholesterol-lowering action of each depends on its ability to lower low-density lipoprotein cholesterol (LDL-C). On the other hand, the molecular structures of the newer statins are not similar and could have an effect on the mechanism of action of the compounds. Differences in metabolism also suggest the possibility of serious drug-drug interactions, and differences in levels of lipid reduction at varying dosages among the statins point to clinical variation as well. PMID- 11383375 TI - What do the statin trials tell us? AB - The results of 5 major placebo-controlled trials evaluating the effects of statins in approximately 31,000 individuals with and without known coronary disease have demonstrated the following: statins reduce the incidence of coronary events, the reduction in relative risk for coronary events increases with the duration of therapy, the reduction in coronary events is proportional to the reduction in low-density lipoprotein cholesterol (LDL-C) levels, and lower LDL-C levels are associated with lower event rates. The studies have also shown that statins are safe and effective in reducing the incidence of coronary events in women, individuals with diabetes, and patients older than 65 years of age and in reducing the risk of stroke and transient ischemic attacks in patients with coronary disease. Finally, studies indicate that statins ameliorate a variety of pathophysiologic processes that are associated with increased risk for atherosclerosis. PMID- 11383376 TI - Surrogate endpoints and newer risk markers in atherosclerosis management. AB - Trials with clinical events as the primary endpoint inherently have poor sensitivity to detect therapeutic effects on plaque stabilization and thrombosis because most plaques that rupture do not cause symptoms. Blood tests or imaging modalities that correlate to the burden or activity of atherosclerosis may provide surrogate endpoints to assess therapeutic efficacy in both clinical trials and clinical practice. For surrogate endpoints to be valid in clinical trials, they must be biologically plausible (i.e., related to the disease process) and altered by therapies that decrease the endpoint for which they are used as a substitute. Examples of surrogate endpoints include progression of coronary disease assessed by angiography, intravascular ultrasound, and other imaging techniques. Risk assessment may be refined and therapy better monitored with blood tests that measure novel markers of atherosclerotic disease. Markers that have been shown to be associated with atherosclerosis include C-reactive protein, intercellular adhesion molecule 1, interleukin-6, and fibrinogen. PMID- 11383377 TI - New therapies on the horizon. AB - Statins have proved to be potent drugs for reducing low-density lipoprotein cholesterol (LDL-C) levels. However, because the response to current statin therapy regimens is not always sufficient to reach defined goal levels, additional drugs to lower LDL-C are needed. New drugs may soon be available to lower LDL-C levels by mechanisms that differ from those of the statins. Among these new agents are a bile acid binding resin, inhibitors of bile acid transport, inhibitors of cholesterol transport, inhibitors of cholesterol esterification, and triglyceride-lowering agents. PMID- 11383378 TI - A new statin: a new standard. AB - In a 2-stage, placebo-controlled, Phase 2 dose-ranging trial evaluating the new statin rosuvastatin in men and postmenopausal women with hypercholesterolemia, the agent was found to reduce low-density lipoprotein cholesterol (LDL-C) levels in a dose-related manner. With each doubling of the rosuvastatin dose (1, 2.5, 5, 10, 20, 40, and 80 mg/day), there was an additional 4.5% reduction in LDL-C. Reductions in LDL-C were statistically significant across all doses and ranged from 35% to 65%. The latter percent reduction surpasses the maximal reductions reported for all other statins when used for monotherapy and suggests that rosuvastatin might enable more patients with hypercholesterolemia to achieve target LDL-C levels. Across all doses, the new statin also had favorable effects on other lipid parameters and was well tolerated, with the incidence of adverse events similar to placebo. PMID- 11383379 TI - [Sleep medicine between science and clinical practice--how long can we avoid the issue?]. PMID- 11383380 TI - Sleep-wake cycles in preterm infants below 30 weeks of gestational age. Preliminary results of a prospective amplitude-integrated EEG study. AB - In the newborn, presence of sleep-wake cycles indicates integrity and maturity of the central nervous system. By spectral EEG analysis and polygraphic recordings subtle variations of EEG background activity and behavioural patterns corresponding to early sleep-wake cycles have been found in preterm infants as young as 27 weeks of gestation. The emergence of sleep-wake cycles at early gestational ages may have a positive predictive value for long-term neurological outcome. Sleep-wake cycles and their significance for later outcome have not been studied in very preterm infants so far. Accordingly, we prospectively investigated maturational changes of EEG activity and sleep-wake cycles in preterm infants below 30 weeks of gestational age using the Cerebral Function Monitor, an amplitude-integrated EEG. We present preliminary data on the emergence of sleep-wake cycles in preterm infants from this ongoing study. Of 100 infants enrolled during a 1-year period, 38 infants without neurological complications were included in the reference group. In this group (mean gestational age 27 weeks), we observed cyclical variations of EEG background activity resembling early sleep-wake cycles at a mean gestational age of 28 weeks and a median postnatal age of 6 days. It is hypothesised that these cyclical variations of EEG background activity may represent switches between thalamo cortical and neo-cortical pattern generators and indicate integrity of central nervous system function. Amplitude-integrated EEG may thus serve as a useful noninvasive test for brain function in preterm infants. PMID- 11383381 TI - [Importance of medical history in diagnosis of respiratory insufficiency in patients suffering from neuromuscular diseases and thoracic deformities]. AB - Patients suffering from neuromuscular diseases and thoracic deformities may develop global respiratory failure during their illness. We wanted to judge clinical parameters and information from the patients' medical history to reliably, quickly and noninvasively diagnose a ventilatory failure. Therefore we evaluated 105 situations with and without mechanical ventilation from 29 patients with indication for noninvasive nocturnal mask ventilation. 6 clinical parameters (e.g. heart rate, oxygen saturation, relative vital capacity), 2 test results (pH and partial pressure of carbon dioxide (pCO2)) and 6 parameters from the patients' medical history (e.g. nycturia, frontal headache in the morning, breathlessness) were investigated. After statistical evaluation we could show a relation between heart rate and pCO2 (Spearman's correlation: r = 0.331, p = 0.001, n = 105; one-tailed significance: r = 0.335, p = 0.038, n = 29). Significant differences between the groups of nycturia incidence indicate a tight relation between the incidence of nycturia and the height of hypercapnia levels (ANOVA--analysis of variance: p = 0.001). Using logistic regression we could show that information regarding medical history, especially nycturia, frontal headache and indrawings, gives important indications for global respiratory failure (sensitivity 97.62-100%, specificity 57.14-76.19%). Pathogenesis needs to be elaborated further. PMID- 11383382 TI - Persistent hypercapnia in children after treatment of obstructive sleep apnea syndrome by adenotonsillectomy. AB - Obstructive sleep apnea syndrome (OSAS) in childhood is frequently in part a consequence of enlarged adenoids and/or tonsils and may lead to hypoxemia and hypercapnia during sleep. Whereas long-term blood gas alterations are well documented in adults, only few polygraphic data are available for children. It was the aim of this study to document blood gas alterations before and after treatment in this population. 9 children with OSAS (6 male, 3 female, median age 5.9 years, range 1.1-13.5 years) were investigated by polysomnography before and after adenotonsillectomy. Prior to intervention most children presented with moderate hypercapnia (ETCO2 mean 44.3 +/- 3.8 mm Hg, ETCO2 maximum 53.2 +/- 5.2) and hypoxemic episodes (oxygen saturation mean 93.2 +/- 3.2%, minimum 74.4 +/- 16.5%). Following adenotonsillectomy subsequent polygraphic investigations displayed normalisation of oxygen saturation (saturation mean 96.1 +/- 0.8%, minimum 90.1 +/- 3.1%). In contrast, moderate hypercapnia in several patients persisted up to five months after treatment (ETCO2 mean 44.9 +/- 2.8 mm Hg, ETCO2 maximum 51.2 +/- 3.6). Persistent hypercapnia most likely reflects an adaptation process of chemosensitivity and respiratory control due to preceding long-term hypercapnia. PMID- 11383383 TI - [Prevalence of sleep disorders in school children between 11 and 15 years of age]. AB - Little is known about sleep disorders in children and adolescents that might affect physical and emotional well-being. Depending on age and size of the cohort group, and differences in questionnaires, prevalence varies between 1-43% in international studies. We examined the prevalence of symptoms characteristic of sleep disorders in school aged children with a questionnaire which allows indication of symptoms by the children themselves. METHODS: An anonymous questionnaire, based on the German Dresden questionnaire, with 22 questions concerning the main symptoms of obstructive sleep apnea syndrome (OSAS), general symptoms of para- and insomnia as well as sociodemographic data, was developed. 332 pupils (age: 11-15 y, mean: 12.75 y; median: 12 y; 56% female, 44% male) in 2 high schools in Vienna were investigated. RESULTS: 28% (n = 93/332) of the examined group reported snoring (the main symptom of OSAS) and/or insomnia (night waking almost every night) or parasomnia (nightmares, night terrors or sleepwalking almost every night). 15% (n = 14/93) of this subgroup reported snoring and para- or insomnia coincidentally. Girls were affected more frequently than boys by nocturnal awakening (79% vs. 56%, p < 0.001) and nightmares (64% vs. 52%, p < 0.01). The snoring group (21% (71/332) of all examined children) was affected more frequently by mouth dryness (16% vs. 4%, p < 0.001), pallor (7% vs. 3%, p < 0.01), night sweats (6% vs. 1%, p < 0.05) and from the following sleep disorders: nightmares (10% vs. 2%, p < 0.01), night terrors (4% vs. 1.5%, p < 0.001), sleepwalking (1.4% vs. 1%, p < 0.05) and nocturnal awakening (16% vs. 5%, p < 0.01). DISCUSSION: Almost every fifth child reports about at least one main symptom characteristic of OSAS. The statistically significant relation between symptoms of OSAS and non-organic sleep disorders shows the necessity of interdisciplinary focusing on sleep disorders. Further epidemiological studies need to be carried out in order to clarify the role of sleep anamnesis in the diagnosis and management of sleep disorders during childhood. PMID- 11383385 TI - Polysomnographic measures in Parkinson's disease: a comparison between patients with and without REM sleep disturbances. AB - STUDY OBJECTIVE: To assess the frequency of rapid eye movement (REM) sleep abnormalities in Parkinson's disease (PD) patients and compare polygraphic sleep measures in those with and without REM sleep disturbances. DESIGN: Polysomnographic recordings of 2 consecutive nights were performed in 45 patients with PD (mean age 65 years, mean Hoehn and Yahr stage 2.2). Twenty patients were treated with dopaminergic drugs, 10 were drug-free for two weeks and 15 had never been treated with L-dopa or dopamine agonists. According to the polysomnographic findings, the patients were divided into those with and without REM sleep abnormalities. Abnormal REM sleep features were defined as REM sleep without atonia (RWA) and REM sleep behavior disorder (RBD). RESULTS: Eighteen (40%) of the PD patients showed either RWA (24%; 6 men, 5 women) or RBD (16%; 6 men, 1 woman). Patients with REM sleep disturbances had a significantly longer duration of the disease (8.3 vs. 3.9 years), a more severe stage of the disease (2.6 vs. 2.0 Hoehn and Yahr stage) and were treated with a higher dosage of dopaminergic drugs (L-dopa, pergolide and bromocriptin). 67% of the patients with normal REM sleep were untreated at the time of the sleep study, but only 39% of those with REM sleep abnormalities. Sleep EEG measures (sleep efficiency, sleep onset latency, sleep period time, relative amounts of sleep stages) for the second night showed no significant differences between both groups apart from a significantly lower sleep period time in PD patients with RWA/RBD. CONCLUSIONS: Abnormal REM sleep features are a frequent finding in patients with PD. The prevalence seems to increase with a longer disease duration. Therefore, a careful follow-up is necessary. A sleep architecture not different from PD patients without RWA/RBD suggests that the underlying abnormality is confined to REM sleep. PMID- 11383384 TI - N-CPAP rejecters--a specific group of noncompliant patients with obstructive sleep apnea syndrome. AB - A large number of studies have been focused on patients with severe obstructive sleep apnea syndrome (OSAS) being non-compliant with N-CPAP therapy, but data concerning patients who reject N-CPAP therapy from the very start, without a day of N-CPAP home therapy, do not exist. The purpose of the study was to determine differences, if any, in the quality of life (QoL) between patients rejecting N CPAP therapy from the start, untreated patients with severe OSAS waiting for N CPAP titration, and controls. Qol was measured in terms of life satisfaction on the basis of the Munich life quality dimension list (MLDL), an instrument for cognitive assessment of elementary components (physical condition, psyche, social life, everyday life) of QoL. Untreated patients (n = 16) with severe OSAS have a significantly worse QoL in the dimension of psyche (p = 0.00022), social life (p = 0.00582) and everyday life (p = 0.01633) than do rejecters of therapy (n = 19). In the dimension of physical condition, no significant difference was seen (p = 0.47138). Compared with controls (n = 113), rejecters of therapy have a significantly lower QoL score in regard of physical condition (p = 0.00014) but not in terms of psyche, social life and everyday life. We conclude that a good QoL and the absence of the stress of suffering is one reason why patients with severe OSAS reject N-CPAP therapy in spite of physical impairment. PMID- 11383386 TI - [Fatigue and stress sensitivity of physicians after 16 hours on duty at the emergency department]. AB - In addition to their 40-hour working week (Mon-Fri, 8 a.m.-4 p.m.) residents at the emergency department of the General Hospital of Vienna have to do approximately six 24-hour duties. The reasons for conducting the present field study were physicians' complaints about tiring night duties. 11 residents (4 women, 7 men; aged between 28 and 43 years, x = 33.5 +/- 4.9 years; working at the emergency department for 4-50 months, x = 31 +/- 20 months) were tested on an ordinary working day at 9 a.m. and midnight. Self-rating concerning sleep duration, perception of stress and workload on the days of the investigations were found to be representative of other prolonged duties. Subjects reported a usual nocturnal sleep duration of only 6-7 hours. Stress was regarded as moderate by most of the volunteers. Blood pressure and pulse rates did not show diurnal changes. Generally, residents felt significantly (p < 0.01) less awake at night than in the morning, but reported only slight vegetative and somatic stress reactions or annoyances as assessed by the Fahrenberg self-rating scale. Interindividual differences were found; residents who had been working at the emergency department for a longer period experienced a more pronounced impairment. Further studies are required in order to objectify a nocturnal decrease in vigilance (by means of computer-assisted EEG) and to evaluate potential performance deficits (by means of psychometric tests). PMID- 11383387 TI - [Sleep-related headaches]. AB - Headaches and sleep disorders are frequent and can be associated with each other. Some headache syndromes are related to certain sleep phases or circadian rhythms. These so-called sleep-related headache syndromes include specific types of migraine, cluster headache, chronic paroxysmal hemicrania, and the hypnic headache syndrome. Except for the latter, they were included in the International Classification of Sleep Disorders and are described in this article. PMID- 11383388 TI - [Nonorganic hypersomnia: epidemiology, diagnosis, and therapy]. AB - Daytime tiredness and daytime sleepiness are frequent complaints occurring in 29% and 14% of the Austrian population. Epidemiological studies demonstrate a high comorbidity between nonorganic hypersomnia and mental disorders. Especially comorbidity with affective disorders increases steadily from the general population over primary to tertiary care settings. Diagnostic criteria of nonorganic hypersomnia have been described in the International Classification of Diseases (ICD-10). Nonorganic hypersomnia can be primary or associated with a number of psychiatric disorders such as reaction to severe stress or adjustment disorders, affective disorders, other functional disorders, tolerance to or withdrawal of CNS-stimulating substances and chronic use of CNS-sedating substances. Diagnostic procedures comprise case history and symptom evaluation, sleep-specific and supplementary investigations. Concerning the latter, this article will focus on sleep questionnaires, vigilance and psychological tests as well as CNS investigations. Therapy of nonorganic hypersomnia rests on 3 pillars: psychological, somatic and pharmacological treatment. In view of the wide variety of psychiatric causes, resulting in a number of therapeutic options, it seems desirable that apart from subjective clinical assessment also objective methods be used in diagnosis and treatment. On the neurophysiological level objective measures can be obtained by means of EEG mapping during the day and polysomnography at night. Different mental disorder patients show different brain activity patterns as compared with normal controls and different classes of psychotropic substances cause different changes in neurophysiological variables. The fact that the changes in electrophysiological brain activity caused by mental disorders are exactly opposite to those induced by the psychotropic drugs used for their treatment suggests a key-lock principle in the diagnosis and treatment of nonorganic hypersomnia. PMID- 11383389 TI - [Excessive daytime sleepiness: etiology, differential diagnosis and diagnostic procedures]. AB - This article gives an overview on frequent causes of excessive daytime sleepiness in clinical practice. Specifically, the insufficient sleep syndrome, sleep disordered breathing, narcolepsy, restless legs syndrome/periodic limb movements in sleep, and circadian disorders causing daytime sleepiness are discussed. Other possible causes including symptomatic daytime sleepiness associated with various medical conditions are mentioned. Different methods to determine daytime sleepiness are discussed in the final section. PMID- 11383390 TI - [Myasthenia gravis and sleep]. AB - In myasthenia gravis respiratory function is often disturbed in the night, especially during REM sleep, despite of normal daytime respiratory function. Nevertheless, nocturnal respiratory problems are rarely diagnosed. Sleepiness, concentration and memory problems can be symptoms of a sleep related breathing disorder. Reports of reduction of REM sleep, memory dysfunction, and detection of acetylcholine receptor (AchR)-antibodies in the cerebrospinal fluid have lead to the hypothesis of a central nervous system involvement in myasthenia gravis. Possible mechanisms are centrally acting AchR-antibodies, unspecifically acting cytokines and hypoxia, possibly the most important influence upon REM sleep reduction and impaired cognitive function. In a patient presenting possible CNS involvement (cephalea, fatigue, concentration and memory problems), a polysomnographic investigation should therefore be performed to detect a sleep related breathing disorder. PMID- 11383391 TI - Activity monitoring in sleep research, medicine and psychopharmacology. AB - Motor activity as a diagnostic parameter has become an important feature in many fields of medicine and psychology. The concept of mobility and immobility implies the assumption that mental and behaviour disorders involve abnormal activity that can be measured to characterise the disorder itself, to diagnose its presence and to document the impact of treatment. In sleep research, activity monitoring by wrist actigraphs has proven its usefulness as an efficient method to assess the rest-activity cycle over long time periods and to estimate sleep-related features such as sleep efficiency and total sleep time. But like many other techniques and devices, activity monitoring has some limitations and drawbacks. This paper describes the basic features of wrist actigraphy in measuring nocturnal and daytime motor activity. PMID- 11383392 TI - [Burnout syndrome in nursing: etiology, complications, prevention]. AB - The scale of burnout and its conditions in nursing are investigated empirically in 28 teams of nurses with 303 staff members in a provincial hospital. 58% of those investigated have more or less strongly pronounced symptoms of burnout. The results show clearly that amongst all variables investigated, the leadership behaviour of ward managers is the variable with the strongest influence on inner burnout in the care profession. Cooperative work relationships, external vocational training courses and team supervision are important in preventing burnout. As expected, there are positive correlations between the working hours and burnout. PMID- 11383393 TI - Perspectives. Waning volunteerism is emergency for rural EMS. PMID- 11383394 TI - Whatever happened to eHealth? AB - Innovations in information technology will continue to transform the health care system to a system that more closely resembles that envisioned in the 1999 and 2001 IOM reports. Healthcare Leadership & Management Report will continue to report over the next few months on practical use of technology to improve the health care system. We are now researching several systems and providers that are using technology to improve their patient care, productivity, bottom line and host of operational concerns. PMID- 11383395 TI - AMA, JCAHO, NCQA release common measures for diabetes care. PMID- 11383396 TI - Had all your shots? PMID- 11383397 TI - Caught between dueling doctors. PMID- 11383398 TI - The problem of under-powering in nursing research. PMID- 11383399 TI - Perceptions about risk for HIV/AIDS among adolescents in juvenile detention. AB - Although progress has been made toward reducing risk-taking behavior among teens, adolescents confined in juvenile detention facilities and youths living in inner cities remain vulnerable. Reaching these populations with appropriate risk reduction strategies continues to challenge health providers and educators. Crucial first steps in the design of relevant programs involve discovering how at risk teens perceive risk and which risks and dangers within their communities occupy their attention. Participants in this study did not identify HIV/AIDS as a primary concern; instead, they described the dangers and risks they encountered in their home neighborhoods. Based on these findings, this discussion addresses the implications for the development of health education programs to empower teens for responsible behavior after release from detention. PMID- 11383400 TI - Outcomes of depression in early adolescents. AB - The purpose of this study was to examine the relationship between depressed mood and depressive symptomatology and the influence of both variables on perceived social support, interpersonal conflict, general well-being, and perceived health status in 144 early adolescents. The subjects responded to instruments measuring the study variables in classroom settings. Two bivariate regression structural equation models were examined using the LISREL 7 computer software program. In the health model, depressed mood had a direct positive effect on depressive symptoms and a direct negative effect on well-being and on perceived health status. Depressive symptoms had a direct negative effect on well-being and on perceived health status. In the interpersonal model, depressed mood had a direct positive effect on depressive symptoms and on conflict and a direct negative effect on social support. Depressive symptoms had a direct positive effect on conflict and a direct negative effect on social support. PMID- 11383402 TI - Families of origin of homeless and never-homeless women. AB - Naturalistic inquiry was used to compare the characteristics of families of origin of homeless women with never-homeless women. The women's experiences in their families of origin were explored during in-depth interviews using Lofland and Lofland's conceptions of meanings, practices, episodes, roles, and relationships to guide the analysis. The two groups were similar with respect to family abuse history, transience, and loss. The never-homeless women had support from an extended family member who provided unconditional love, protection, a sense of connection, and age-appropriate expectations, as contrasted with homeless women who described themselves as being without, disconnected, and having to be little adults in their families of origin. The experience of family love and connection seemed to protect never-homeless women from the effects of traumatic life events in childhood. These findings provide support for the influence of a woman's family of origin as a precursor to homelessness. PMID- 11383401 TI - Uprooting and resettlement experiences of South Asian immigrant women. AB - The purpose of this descriptive qualitative study was to examine and understand the challenges faced by elderly women from India who immigrated to Canada. Ten women were interviewed about their experiences with immigration and resettlement. The analysis of interview data involved iterative process, through which four themes were identified. These themes were isolation and loneliness, family conflict, economic dependence, and setting in and coping. The participants experienced loss because of changes in traditional values and lack of social support. Because the participants could not manage resettlement on their own, personal independence was not very important. Interdependence for the attainment of emotional security and social rewards was more desirable. Health care professionals must take into account the nature of stress and impact of these experiences on health of older immigrant women. PMID- 11383403 TI - Common patterns of person-environment interaction in persons with rheumatoid arthritis. AB - This study's purpose was to identify and describe common patterns of person environment interaction in persons with rheumatoid arthritis. Newman's propositions about pattern provided the theoretical foundation, and a combination of quantitative and qualitative techniques were used to recognize pattern. Sixty six adults diagnosed with rheumatoid arthritis completed questionnaires that assessed characteristic ways of functioning. Iterative cluster analysis of the data identified five distinct groups of individuals with similar response profiles. Content analysis of each group's characteristics produced a core theme representing the nature of the common pattern. The finding of multiple, distinct common patterns of person-environment interaction within the population of persons with RA is consistent with findings in other populations. Further study of the common patterns, in relation to health outcomes, has potential for increasing knowledge about the sources of different health experiences among persons with the same chronic illness. PMID- 11383404 TI - How do you develop a cost management system? PMID- 11383405 TI - Job sharing as an employment alternative in group medical practice. AB - Although physicians discuss quality-of-life and employment issues with their patients, they often fail to consider flexible scheduling and reduced employment options to lessen their own job stress. We examined one of these options by surveying two community-based, private practice groups with a combined 13-year experience with job sharing. We found that a majority of respondents rated job sharing as successful, and most wanted it to continue. Job sharers derived considerable personal benefit from the arrangement and had significantly more positive attitudes toward work than full-time physicians. Job sharing appeared to have little impact on practice parameters. Dependability, flexibility and willingness to cooperate were the most important attributes in choosing a job sharing partner. Job sharing is an employment alternative worth exploring to retain physicians in medical group practice. PMID- 11383406 TI - Developing a physician needs assessment. AB - The physician needs assessment allows a health care organization to figure the number and mix of physicians it will need, assisting in targeted, realistic recruitment efforts. It forms a key piece of your organization's strategic plan, lays out the current specialty mix, helps justify recruitment plans to your medical staff and the physicians you are recruiting, and can maintain your practice's good standing with regulatory agencies. PMID- 11383407 TI - Leadership survey. An evaluation of health care executives' challenges. AB - Locating and keeping employees represents one of the greatest challenges facing health care leaders today. This is a key finding of the third Leadership Survey of executives in physician practices, managed care organizations and hospitals. The survey is sponsored by the Medical Group Management Association and Hospitals & Health Networks magazine. Other significant results: Practices put the most emphasis on teamwork, training and staff development as methods to combat labor shortages; practice executives count adequacy of reimbursements and physician productivity as top leadership challenges, along with the availability of qualified workers; practices choose print advertising and the addition of new products and services as the best ways for them to build market share. PMID- 11383409 TI - Which managed care contract(s) should you sign? AB - Contracts are a basic responsibility in running a medical practice--so much so that it's inappropriate to pass those responsibilities to someone else. Managers must at least understand the basic terms and conditions of the contract and the reimbursement their practices will receive. This article is written from the perspective of a privately owned medical practice and addresses only discounted, fee-for-service managed care contracts. PMID- 11383408 TI - A new paradigm in academic health centers: productivity-based physician compensation. AB - This article describes the challenge of designing an incentive-based compensation program for a large group of academic pediatricians in the Division of General Pediatrics at the University of Michigan Health System. The program is based on an incentive system that measures performance in clinical care, education and research. Faculty members' salaries arise from five components: base, clinical incentive, academic supplement, administrative differential and teaching credit. PMID- 11383410 TI - Credibility in medicine: a physician's perspective. PMID- 11383411 TI - Liability insurance offers more than it used to. PMID- 11383412 TI - "They treated me like a criminal". PMID- 11383414 TI - E-health gurus still expect to transform your world. PMID- 11383413 TI - "Doctor, I know there's something wrong". PMID- 11383415 TI - Shaping your profile on the Web. PMID- 11383416 TI - "All the good doctors always leave". PMID- 11383417 TI - A medical crisis: who'll care for your patients? PMID- 11383418 TI - Spot the safety hazards in this office--and yours. PMID- 11383419 TI - How do we get back our humanity? PMID- 11383420 TI - These programs focus on relationships. PMID- 11383421 TI - Quality measures for the emergency services. PMID- 11383422 TI - The standards for emergency surgical services. AB - Gross underfunding of the National Health Service in England and Wales results in too few beds and operating theatres and too few nurses and doctors. Thus, standards of surgical care, particularly for emergencies, are compromised. The service requires sufficient senior and trainee surgeons to meet the needs of specialization, working together in an acceptable surgical rota which enables both dedication to emergency admissions and continuity of care. Calculation of local manpower needs demands an understanding of the acceptable workloads for operating and outpatient activity and assessment of the NHS and private surgical work carried out in the area. For general surgery and trauma and orthopaedics this equates to 1 consultant for 30,000 population. Emergency surgical services require the presence on site of all the core specialties, including sufficient fully staffed intensive-care, high-dependency and coronary care beds to ensure their availability for emergency admissions together with 24-hour-staffed dedicated emergency operating theatres. PMID- 11383423 TI - Quality measures for the emergency obstetrics and gynaecology services. AB - The maternal mortality rate was the first measure of quality in the obstetric services. It is a crude indicator but is still used for international comparisons. In the UK, confidential enquiries into maternal and perinatal deaths produce recommendations the implementation of which is not well audited as yet. 'Near misses' are harder to define but are audited in individual units. Standards drawn up by the Central Negligence Scheme for Trusts could potentially promote improvements. The Royal College of Midwives and the Royal College of Obstetricians and Gynaecologists have jointly published standards of care in labour wards. Gynaecological standards are less well developed but should evolve as NHS audit improves. PMID- 11383424 TI - Standards in accident and emergency medicine. AB - There are numerous standards currently available that relate to accident and emergency medicine. Some of these relate to organizational structure; others are clinical and relate either to the process of care or to outcomes. Few, if any, deal explicitly with the dimensions of quality mentioned in recent white papers about the NHS. It is suggested, to maximize the effect standards have on care, that they should be developed for existing technologies not just for novel ones, rigorously developed and effectively disseminated and implemented, formally evaluated after their introduction and mutually compatible. PMID- 11383425 TI - What are the standards for the emergency anaesthetic services? AB - Anaesthetists provide services throughout acute hospitals in areas such as the delivery floor and the intensive therapy unit as well as working in their traditional role in the operating theatre. Consensus standards of the number of staff needed to provide a satisfactory level of acute anaesthetic services, their qualifications and experience and the resources they require have been produced by a number of organizations. It is probable that many small and medium-sized district general hospitals will be unable to meet these standards without changes to traditional UK staffing structures. PMID- 11383426 TI - Setting standards for pathology service support to emergency services. AB - Quality standards have been established in two key areas of pathology directly relevant to standards for the provision of emergency medical services. First, there is a national scheme for accreditation of laboratory services--Clinical Pathology Accreditation (UK) Ltd (CPA)--which has been in formal operation since 1992 and currently covers about 80% of all UK laboratories. Secondly, guidelines have been issued by the Joint Working Group on Quality Assurance (JWGQA) on the support to any point-of-care (near patient) testing facilities. Point-of-care testing (POCT) is increasingly popular in emergency areas, where the availability of faster test results is expected to expedite diagnosis and treatment. When laboratory services are not accredited or POCT equipment and its usage are outside laboratory supervision, there should be concerns that quality standards for pathology service support of the emergency services are not being met. PMID- 11383427 TI - Evaluating emergency services activity at the health district level. AB - We do not have good information on the incidence and prevalence of emergency conditions nor is there good research evidence on the best ways of meeting these. There are, however, some indicators for evaluating emergency services activities and we have a good framework from Donabedian for evaluation, and the important dimensions of quality specified by Maxwell. The range of emergency services covers primary care, community crisis care, ambulance services, hospital services (accident and emergency [A&E] department, inpatient, critical care), laboratory (blood supplies, tests), social services, and public health. There are about eight main sources of current indicators. Unfortunately the availability, timeliness, and quality of these indicators is variable. A new development is situation reporting on emergency pressures ('Sitreps'). This provides a fortnightly and sometimes daily picture of current emergency activity as measured by key indicators such as the number of delayed discharges from hospital, the number of cancelled operations, and the number of medical inpatients outlying on other wards. 'Sitreps' was particularly helpful in handling emergency activity at the new millennium period. We need to specify a comprehensive, valid and easily collectable data set for assessing the quality of emergency services. This would include better ways of forecasting for early warning purposes. This could be done by monitoring the incidence of absenteeism, the sale of over-the-counter drugs, and the number of deaths in nursing homes. PMID- 11383428 TI - Clinical governance in pre-hospital care. AB - This article seeks to discover and recognize the importance of clinical governance within a new and emerging quality National Health Service (NHS) system. It evaluates the present state of prehospital care and recommends how change, via clinical governance, can ensure a paradigm shift from its currently fragmented state to a seamless ongoing patient care episode. Furthermore, it identifies the drivers of a quality revolution, examines the monitoring and supervision of quality care, and evaluates the role of evidence-based practice. A frank and open view of immediate care doctors is presented, with recommendations to improve the quality of skill delivery and reduce the disparity that exists. Finally, it reviews the current problems with pre-hospital care and projects a future course for quality and patient care excellence. PMID- 11383429 TI - Objective standards for the emergency services: emergency admission to hospital. AB - The primary objectives of the emergency services are to minimize early mortality and complications, although longer-term morbidity, quality of life and late mortality may also be influenced by early actions. Evaluation of the emergency services and demonstration of quality need to reflect these objectives by appropriate choice of outcome measures. This brief review of leading measures of quality in emergency admissions discusses population-based 30-day mortality, after examining some limitations of 'hospital mortality', incidences of complications, which can be more sensitive measures of quality of care when mortality rates are low, and a role for audit and management, when relationships between process and outcome are clear. As an example, the UK study of urgent admission to hospital, on behalf of the Clinical Standards Advisory Group, showed wide variation between sampled hospitals in timeliness of early clinical actions and a statistically significant association between timeliness and 30-day survival. The review also discusses capacity, a necessary requirement for a quality service, and operations research/queuing theory to facilitate management of capacity/resources to meet fluctuating demands. The NHS should be able to plan for seasonal needs. PMID- 11383430 TI - Standards of access and quality in primary care. AB - This paper reviews recent changes in the provision and organization of primary care in the UK. Access and availability are of prime importance to patients, particularly when medical care is sought urgently, and are therefore important elements of quality. The paper also discusses the possible impact of further changes in the delivery of primary care, including overlapping responsibilities of general practice and accident and emergency (A&E) departments and the role of NHS Direct, a telephone advice service. PMID- 11383431 TI - The interface between primary and secondary care. AB - The interface between primary and secondary care in the UK has been affected by a number of recent changes, particularly in provision of out-of-hours care and advice. This paper reviews some current measures of healthcare quality and argues that many do not adequately measure contributions in primary care. To overcome these deficiencies the Royal College of General Practitioners (RCGP) has published guidelines on issues of quality in primary care. PMID- 11383432 TI - Planning and policy for emergency services: four fallacies, two problems and some possible solutions. AB - This paper examines some of the misapprehensions that have often underpinned the planning of accident and emergency services in the UK. Accident and emergency (A&E) is not a homogenous group of activities and the different components that make up the service should be planned separately. This planning needs to be accompanied by some significant redesign to meet growing patient expectations. In particular, there is a major challenge for services to offer local access in an environment in which acute care is increasingly centralized. PMID- 11383433 TI - What will be the costs of a quality emergency service? AB - The Clinical Standards Advisory Group report on urgent and emergency admissions to hospital identified several important issues regarding emergency care. This paper examines what we know about the costs of a quality emergency service. Estimating the cost of a quality emergency service is not yet possible, but ongoing and future research should be capable of highlighting the costs of important service improvements. It must, however, be recognized that such work by itself will not automatically lead to increased funding within emergency care. Further consideration must be given to the funding arrangements for emergency care if long-term improvements are to be secured. PMID- 11383434 TI - Improving quality measures in the emergency services. AB - A large and continuing increase in medical emergency admissions has coincided with a reduction in hospital beds, putting the acute medical services under great pressure. Increasing specialization among physicians creates a conflict between the need to cover acute unselected medical emergencies and the pressure to offer specialist care. The shortage of trained nursing staff and changes in the training of junior doctors and the fall in their working hours contribute to the changing role of the consultant physician. The organization of the acute medical service is of paramount importance and requires multi-disciplinary teamwork on an admissions unit with full support services. Excellent bed management is essential. There must be guidelines for all the common medical emergencies and all units must undertake specific audits of the acute medical service. Continuing professional development (CPD) and continuing medical education (CME) should reflect the workload of the physician; that is, it must include time specifically focused on acute medicine and general (internal) medicine, as well as the specialty interest. PMID- 11383435 TI - Why linguistics needs the cognitive scientist. PMID- 11383436 TI - Why psychiatry and cultural anthropology still need each other. PMID- 11383437 TI - Edward Sapir's thought experiment in the interdisciplines of cultural anthropology and psychiatry. PMID- 11383438 TI - Why cultural anthropology needs the psychiatrist. 1938. PMID- 11383439 TI - Sapir's vision of culture and personality. AB - For Edward Sapir the concept of culture was a reification of processes that were rooted in individuals' personality and psychology. Sapir suggested that psychiatry's focus on individual biography and pathology gives it unique relevance for social science efforts to understand the mechanisms of cultural transmission and transformation. As a discipline that must integrate culture and biology in theory and practice, psychiatry can provide a corrective to the extremes of biological or cultural reductionism. Although mainstream psychiatry has largely abdicated the role it once had in the social sciences, the interdisciplinary field of cultural psychiatry may meet some of Sapir's hopes. Recent work in cultural psychiatry is centrally concerned with illness narratives that arise from the interaction of personal and collective meaning. Illness narratives may serve individual defensive functions, position individuals in a social world, and help to maintain overarching cultural formations. They also may challenge or subvert existing cultural meanings and create new forms of discourse. The close analysis of how cultural and individual meanings interact that is provided by cultural psychiatry has much to offer the wider field of cultural anthropology. PMID- 11383440 TI - Interview with Robert Cohen and Donald Burnham. Interview by Jon Frederickson. PMID- 11383441 TI - Trauma and extended separation from family among Latin American and African refugees in Montreal. AB - Although the vast majority of refugees have suffered trauma and extended separation from their families in exile, little is known about the interactions between these two types of experience. The qualitative and quantitative analyses of data gathered from 113 refugees from Latin America and Africa suggest that the joint occurrence of trauma and separation has a significant impact on emotional distress and confirm that the family plays a key role as an anchor of emotion and identity. PMID- 11383442 TI - The trauma story: a phenomenological approach to the traumatic life experiences of refugee survivors. PMID- 11383443 TI - Forced relocation: a family researcher/clinician's perspective. PMID- 11383444 TI - Do children cope better than adults with potentially traumatic stress? A 40-year follow-up of Holocaust survivors. AB - Anecdotal reports suggest that child survivors of the Nazi persecution are functioning well as adults. Ratings of their parents by a randomly selected community sample of young adult Ashkenazi Jews on a scale that measured Schizoid, Paranoid, Depressive/Masochistic and Type A/Normal Aggressive symptoms permitted verification of these reports. Among the parents were groups who were children, adolescents, or young adults in 1945, at the end of World War II. Child-survivor parents did not differ from native-born parents on these measures 40 years later, whereas, consistent with the empirical findings of others, survivors who were adolescents or young adults at the end of the war manifested more paranoid and depressive/masochistic symptoms than native-born parents. To explain this possible greater long-term resilience among those who were child survivors, reference is made to later caretakers, endowment, cognitive and social development, and psychodynamics. PMID- 11383446 TI - The psychological uses of fiction. PMID- 11383445 TI - Nocturnal hallucinations in ultra-orthodox Jewish Israeli men. AB - Hallucinations that occur predominantly at night are reported in 122 out of a sample of 302 ultra-orthodox Jewish Israeli men referred for psychiatric evaluation. Demographic data and the content of a semistructured interview in 302 ultra-orthodox Jewish young men seen over a 10-year period in Jerusalem were evaluated retrospectively by two researchers. Of the 302 subjects, 122 reported hallucinations predominantly at night, 23 reported hallucinations with no diurnal variation, and 157 did not report hallucinations. Most of those with nocturnal hallucinations were in their late teens, were seen only once or twice, were brought in order to receive an evaluation letter for the Army, and had a reported history of serious learning difficulties. The nocturnal hallucinatory experiences were predominantly visual, and the images were frightening figures from daily life or from folklore. Many of the subjects were withdrawn, monosyllabic, reluctant interviewees. Ultra-orthodox Jewish beliefs include a belief in demons, particularly of dead souls, who visit at night. This cultural group's value on study at Yeshivas away from home places significant pressure on teenage boys with mild or definite subnormality, possibly precipitating the phenomenon at this age in this sex. Although malingering had to be considered as a possible explanation in many cases owing to the circumstances of the evaluation, short-term and long term follow-up on a limited sample allowed this explanation to be dismissed in a significant number of cases. We suggest therefore that nocturnal hallucinations are a culture-specific phenomenon. PMID- 11383447 TI - [The scientific career of V. M. Dil'man]. AB - The life and research career of Prof. V.M. Dilman, who would have been 75 in July, 2000, are briefly outlined. V.M. Dilman had been on the staff of the N.N. Petrov Research Institute of Oncology for many years. He contributed much to the development of oncology, endocrinology and gerontology. Among the basic features of the ontogenetic model of development and aging put forward by V.M. Dilman are the age-associated emergence of hormonal-metabolic factors of increased risk for cancer and relevant measures to be taken to retard this process. PMID- 11383448 TI - [Oncologic diseases in menopause: causes and methods of prevention]. AB - Cancers of the breast, endometrium, ovary cervix and colon clearly have an important impact on women's health worldwide, in terms of their burden on incidence and mortality. Our understanding of causes of these cancers is far from complete. However, the present state of knowledge does give clues to biological mechanisms and for continued research. Presently, some prospects for prevention may be proposed. Changes in life styles regarding physical activity, diet and perhaps reproductive behavior, and also use of safe hormonal treatments, are possibilities for primary prevention. For secondary prevention, methods are available for screening early lesions in the breast, uterine cervix and colon that may enable curative treatment and reduction of mortality. Urgent topics for research include the identification of risk factors early in life and meaningful aspects of diet and life style that can be modified, and finally the investigation of chemical agents to prevent, for instance, breast cancer. PMID- 11383449 TI - [Age, environmental factors, and hormonal carcinogenesis]. AB - In this paper modern concepts of hormonal carcinogenesis are discussed. Distinguishing of the latter into 2 principal types (promotional and genotoxic) is supported. The role of age, aging and effect of environmental factors in the choice of hormonal carcinogenesis type (as well as for the creation of "estrogenic effect switching/overtargeting phenomenon") is underlined. Mentioned phenomenon is manifested by the weakening of endocrine (hormonal) effects of estrogens together with intensification of genotoxic (DNA-damaging) effect of their metabolites. Conclusion is made that both the conditions favoring hormone dependent tumors development and the direct mechanisms of hormonal carcinogenesis may be considerably modulated on different stages of ontogenesis and by the influence of environmental factors that is to be taken into considerations while choosing methods for oncological diseases prevention. PMID- 11383450 TI - [Genetic polymorphism and variability of steroid hormone metabolism: connection with risk of developing breast neoplasms]. AB - Endogenous estradiol is synthesized in the ovarian theca cells of premenopausal women or in the stromal adipose cells of the breast of postmenopausal women and in minor quantities in peripheral tissue. These cells, as well as breast cancer tissue, express all the necessary enzymes for this synthesis: CYP17, CYP11a, CYP19, hydroxysteroid hydrogenase, steroid sulphatase as well as enzymes further hydroxylating estradiol such as CYP1A1, CYP3A4, CYP1B1. Polymorphisms in these enzymes may have a possible role in the link between environmental estrogens and hormone-like substances and the interindividual risk of breast cancer. PMID- 11383451 TI - [Role of oncogenic G-proteins in development of endocrine neoplasms]. AB - New data available in the literature on the role of oncogenic heterotrimeric GTP proteins (G-proteins) in endocrine tissue carcinogenesis have been analyzed. A conclusion is made that stimulatory G-proteins coded by the gsp-oncogene are a major factor of the development of some pituitary and thyroid tumors. However, the role of the other oncogenic G-proteins in tumorigenesis remains unclear. PMID- 11383452 TI - [Hormones, autocrine and paracrine regulators of tumor growth in osteosarcoma]. AB - The data on 300 patients, aged 14-56, treated at the Center's Clinic have been evaluated. They include findings on androgen metabolism, basal level of secretion of sex steroid hormones in blood serum and their receptors in tumor, blood concentrations of sex steroids, binding globulin and pituitary hormones, arachidonic acid metabolism, expression of epidermal growth factor and its receptors and ligands, soluble Fas-antigen, vascular endothelial growth factor as well as calmodulin and cAMP levels in sarcoma. New trends of research in pathogenetic methods of treatment of osteosarcomas used in practice of oncology have been identified. Among them are androgen metabolism regulation, correction of the cyclooxygenase pathway of arachidonic acid metabolism, expression of epidermal growth factor and its ligands and neoangiogenesis in tumor. PMID- 11383453 TI - [Hormonal and molecular biological factors in pathogenesis of prostate cancer]. AB - Moleculo-genetic pathways of development and progression of prostate cancer have been studied. In the norm, paracrine regulation of the glandular secretory epithelium are predominant due to the influence of hormonal and protein factors of the stroma. The altered prostatic epithelium becomes independent from the stroma and androgens and, consequently, prone to metastasizing, following activation of protooncogenes (growth factor genes), inactivation of gene suppressors, hyperexpression of certain growth factors and apoptosis inhibitor Bcl-2, and inactivation of androgen receptor. In inoperable prostate cancer, palliative treatment should include antiandrogen therapy, inhibition of growth factors and activation of apoptosis. Prevention and medication for prostate cancer targeting apoptosis, growth factors and androgens should be based on recent achievements in experimental genotherapy and selective use of proliferation inhibitors and apoptosis activators which are to be fed to the gland via viral and non-viral vectors. Molecular-biological markers of risk for cancer, its early detection, prognosis of clinical course and effectiveness of treatment are discussed. A modality for laboratory diagnosis and complex treatment of prostate cancer offering maximum survival so far has been suggested. PMID- 11383454 TI - [Aromatase and breast cancer]. AB - Aromatase activity may be detected using in vitro and in vivo techniques in most breast cancers and mammary adipose tissue. This activity makes a variable contribution to endogenous estrogens within the breast and in many cases represents the major source of these hormones. Such local biosynthesis may maintain the growth of some hormone-dependent tumors. The factors which regulate aromatase activity within the breast are not defined but are likely to include growth factors and cytokines which may be produced by breast tissues so that autocrine and paracrine loops may exist. Estrogen biosynthesis within the breast, like other peripheral systems, appears sensitive to classical aromatase inhibitors and the new generation of drugs are capable of profoundly blocking the activity and markedly reducing endogenous estrogen levels; in turn these endocrine effects are translated in dramatic anti-tumor influences in hormone dependent breast cancer. PMID- 11383455 TI - [Potential for using aromatase inhibitors for preventing breast cancer]. AB - Substantial evidence supports the concept that estrogens cause breast cancer in animals and in women but the precise mechanism is unknown. The most commonly held theory is that estrogens stimulate proliferation of breast cells and thus statistically increase the chances for genetic mutations which could result in cancer. Another theory is that estrogen metabolism generates oxygen-free radicals and quinones which produce both stable and unstable DNA adducts. Both result in genetic mutations which accumulate and could ultimately cause cancer. A major criticism of the latter hypothesis is that breast tissue contains insufficient concentrations of estrogen for accumulation of genotoxic metabolites. Our hypothesis is that breast tissue estrogen levels, as a result of in situ synthesis, are much higher than previously thought. With sufficient amounts of aromatase in breast tissue, enough estradiol as substrate should be available to allow formation of substantial amounts of genotoxic metabolites. We postulate that aromatase overexpression may in this way cause breast cancer. As evidence supporting this concept, four animal models of aromatase overexpression and either breast cancer or pre-malignant lesions have been described. We have provided evidence that normal breast tissue can make estrogen and that certain stimulatory compounds can increase aromatase activity in the breast by nearly 10,000-fold. If our concepts are correct, it might be possible to prevent breast cancer by blocking the aromatase enzyme. Aromatase inhibitors might be more effective than antiestrogens in preventing breast cancer because of their dual role to block both initiation and promotion of breast cancer. PMID- 11383456 TI - [Modern approaches to hormone therapy of breast cancer as a reflection of pathogenesis of the disease]. AB - Experimental and epidemiological studies have pointed to a major role of estrogens in the pathogenesis of human breast cancer. The Oxford meta-analysis (1998) once again confirmed the efficacy of antiestrogens (tamoxifen) as adjuvant therapy. We need to know whether the new non-steroid antiestrogens (idoxifen, droloxifen and TAT-59) and selective estrogen receptor modulator (raloxifen), whith preclinical characteristics better than those of tamoxifen will be more efficient clinically. Large-scale trials to compare the new drugs with tamoxifen are under way. Faslodex, a pure antiestrogen, looks highly promising, too. Zoladex, a luteinising hormone-releasing hormone agonist, is looking as a better choice than ovariectomy or irradiation of the pelvis for ovarian ablation in premenopausal breast cancer. New aromatase inhibitors are more efficient than progestins and much safer than aminoglutethimide. It has been shown recently that these inhibitors keep metastatic breast cancer at bay longer, and with longer survival. The non-steroid inhibitors (anastrozole and letrozole) and the steroid oral drug exemestane are undergoing clinical trials as means of adjuvant treatment of breast cancer. The trial of arimidex and tamoxifen administered alone or in combination (ATAC) is unique since it is using a combination of tamoxifen and an aromatase inhibitor (anastrozole). New methods of endocrine therapy have resulted in less toxic and more convenient procedures. Also, longer therapeutic effects and survival are becoming more apparent. PMID- 11383457 TI - [Molecular-genetic determinants of the neoplastic process and state-of-the-art treatment of patients with uterine leiomyoma]. AB - Present-day evidence on uterine leiomyomata has pointed to progesterone and its receptors as a key factor in the mechanisms of auto- and paracrine influences on tumor development and growth. Treatment was determined by a correlation between tumor size and stage of molecular-genetical disorders. A clinico-genealogical study of familial predisposition to uterine leiomyoma, particularly in patients from accumulated disease families contributed to the potential of early detection of tumor and timely effective correction for preservation of reproductive function. PMID- 11383458 TI - [Level of estrogen receptors in breast tumors: role of ethnic factors]. AB - Estrogen receptor levels have been assayed in breast cancer patients. A significant increase in receptor-negative tumors was found in Kazakh menopausal women. The proportion of receptor-positive tumors correlated inversely with tumor progression, irrespective of ethnicity. A significant rise in the incidence of receptor-negative tumors was recorded in Kazakh females with breast cancer stage III. PMID- 11383459 TI - [Correlated connection of endocrine function of thymus and other internal secretion glands at stages of developing uterine choriocarcinoma]. AB - The mechanisms of endocrine and immune system disorders in normal and disturbed pregnancy as well as in chorionocarcinoma, hydatid mole, metastatic and non metastatic chorionocarcinoma have been investigated. Relevant disorders were chiefly caused by the endocrine function of the thymus and gonadotropins from the trophoblast, which made a powerful impact on the periphery of hormonal and immunological homeostasis. Unlike normal pregnancy, gestational trophoblastic disease involved changes in the nature, direction and degree of correlations between endocrine formations, which in turn determined the pathogenetic pattern of the disease. PMID- 11383460 TI - [Effect of neoadjuvant therapy with neovir on the level of steroid hormone receptors in endometrial cancer tissue]. AB - The effects of neoadjuvant therapy with neovir, provera and their combinations on endometrial tumor morphology, progesterone (PR) and estradiol (ER) receptor levels were studied in 50 patients, aged 40-78, with primary tumors stage I-III, and without concomitant pathology. A 19-day course of neovir did not alter tumor morphological status but was followed by a significant rise in PR and ER concentrations. After neoadjuvant therapy in combination with provera, which generally lowers cytoplasmic receptor levels, mean values of receptors dropped in PR+ tumors while, in PR- tumors, they increased significantly. Our findings suggest that neovir may be used in clinic to stimulate endometrial tumor sensitivity to specific hormonal therapy, particularly, for hormone-independent neoplasia. PMID- 11383461 TI - [Role of phosphatidylinositol signalling path in developing hormonal resistance in tumor cells]. AB - Phosphatidylinositol 3-kinase (PI3K) is a key regulatory protein which is responsible for anti-apoptotic signal transduction regulating cell survival during exposure to damaging factors. The report deals with the role of the PI3K signaling pathway in regulating cellular response to hormones and, particularly, in development of resistance as a result of long-term exposure of cells to steroid cytostatic hormones. In our study, even a short-term exposure of transformed fibroblasts of hamster (line 2PK) resulted in an activation of main PI3K effectors (MAP-kinase and protein kinase B (PKB)) which appeared against the background of hormone-induced inhibition of cellular growth. A long-term (3 months) cell culturing with dexamethasone was followed by formation of subpopulations of cells which were refractory to the growth inhibition by hormone and were characterized by high levels of activity of PI3K, MAP-kinases and PKB. Activation of PI3K and PI3K-dependent enzymes correlated with enhancement of synthesis of c-jun, a component of the AP-1 transcriptional factor, was observed both in short- and long-term application of dexamethasone. We believe that, during long-term exposure of cells to cytostatic hormones, continuous activation of PI3K and PI3K-dependent transcriptional factors may result in a significant restructuring of intracellular signal pathway, and, finally, constitutive PI3K signal pathway and partial overcoming the proliferative block by cells. PMID- 11383463 TI - Why don't office pediatricians use more technology? An historical perspective. PMID- 11383462 TI - [Bcl-2 tumor suppressor as a possible mediator between genotoxic stress and disturbance in hormonal homeostasis]. AB - Overexpression of tumor suppressor gene bcl-2 plays an important role in cellular resistance to apoptosis caused by various factors including glucocorticoids. In this study, the role of bcl-2 in glucocorticoid-mediated negative feedback regulation has been investigated. Transient transfection of bcl-2 into murine corticotrope AtT-20 cells resulted in significant resistance of proopiomelanocortin gene expression and ACTH secretion to down-regulation by dexamethasone. Bcl-2 revealed its activity mostly in the presence of saturating concentrations of dexamethasone (100 nM-1 mM). Overexpression of bcl-1 interfered with the receptor-mediated glucocorticoid activity and appeared to be relatively specific towards expression of proopiomelanocortin/ACTH. The data suggest a novel function of bcl-2 as a factor capable of regulating hormonal homeostasis. Thus, bcl-2 may be involved in hormonal and metabolic response to genotoxic stress, the phenomenon which was earlier defined as carcinogenic aging. PMID- 11383464 TI - After-hours telephone care: options for the pediatrician. PMID- 11383465 TI - Cost-efficient telephone care during pediatric office hours. PMID- 11383466 TI - Creating an after-hours telephone triage system for office practice. PMID- 11383467 TI - Applying clinical guidelines to pediatric practice. PMID- 11383468 TI - Efficient vaccination practices. PMID- 11383469 TI - The coming of age of the electronic office. PMID- 11383470 TI - Resident's column: technology in pediatric residency. PMID- 11383471 TI - Test designs to assess the influence of soil characteristics on the toxicity of copper and lead to the oligochaete Enchytraeus albidus. AB - Statistical test designs were used to identify and quantify the soil parameters important for determining the ecotoxicological effects of copper and lead to the potworm Enchytraeus albidus. The application of a Fractional Factorial Design revealed that the acute toxicity of copper and lead to E. albidus can vary over more than two orders of magnitude depending on the physico-chemical characteristics of the (artificial) soils. The differences in metal ecotoxicity were mainly determined by pH and organic matter content or cation exchange capacity (CEC). Using a Central Composite Design, models were developed describing the ecotoxicity of copper and lead to E. albidus as a function of these parameters. To validate the developed response surface models, two field soils and the standard artificial soil prescribed by OECD were spiked with copper and lead and the acute toxicity to E. albidus was assessed. These validation experiments confirmed that the toxicity of copper and lead could be predicted using information on the total metal concentration, the pH and the CEC of the soil with toxicity decreasing with increasing pH and CEC. PMID- 11383472 TI - Responses of chub (Leuciscus cephalus) populations to chemical stress, assessed by genetic markers, DNA damage and cytochrome P4501A induction. AB - Indicators of effects at the population level (genetic variation using allozymes) and early indicators of pollution (EROD activity and DNA strand break formation) were analysed in chub (Leuciscus cephalus) living in weakly and heavily contaminated stations of the Rhone River watershed. The genetic erosion was mainly detected in a fish population living in a contaminated small river system, through modifications in allelic and genotypic frequencies for PGM-2 locus and could be linked to a genetic bottleneck and to the reduced gene flow from upstream unable to maintain or restore the genetic diversity. In a contaminated large river system, the genetic diversity for PGM-2 and other loci was maintained and was probably the consequence of a high gene flow from upstream, linked to a sustained drift of larvae and juveniles in the system. A convergent increase of the frequency of the 90 allele at PGM-2 was observed in two contaminated stations compared with the reference station, this trend being confirmed on a more extensive geographic scale over the Rhone River basin. A high level of EROD activity was detected in both contaminated sites but only the fish in the large river system showed a significant DNA damage level compared to the reference population. The low DNA damage level and high hepato-somatic ratio characterized the impacted population of the small river system and could be associated to a chronic high-level exposure of fish to pollutants which selected individuals exhibiting a high level of DNA damage repair. In the two contaminated systems, some genotypes at the PGM-2 and EST-2 loci showed a low level of DNA damage and/or a high EROD activity and may be considered as being tolerant to pollutants. A higher tolerance of the most heterozygous fish was also detected in the contaminated large system and confirmed that a high level of heterozygosity may be necessary for survival in such a system. PMID- 11383473 TI - The potential use of the South African river crab, Potamonautes warreni, as a bioindicator species for heavy metal contamination. AB - In 1995, preliminary water and sediment analyses of the river bed and burrow sediments from 9 locations along the Mooi River, NW Province, South Africa had shown cadmium concentrations up to 0.009 mg l-1 +/- 0.003 and up to 0.33 and 0.89 weight % with scanning electron microscopy (SEM) and x-ray microanalysis. Samples of the adult river crab (Potamonautes warreni) were collected from the Mooi River at Noordbrug (26 degrees 40'S/27 degrees 05'E), 1 km north of Potchefstroom Town, and exposed to 0.2 or 2.0 mg Cd2+ l-1 in situ to determine tolerance, uptake and bioaccumulation of cadmium. Using flame atomic absorption spectroscopy (FAAS) the gills, haemolymph and digestive gland of naturally exposed P. warreni showed wet mass values of 0.74 +/- 0.27 microgram Cd2+ g-1, 0.007 +/- 0.007 microgram ml-1 and 0.12 +/- 0.09 microgram g-1 respectively. The tolerance of crabs to aqueous Cd reached its limit (ET50 = 42 hours) at 2.0 mg l-1 aqueous Cd exposure. At an exposure to 0.2 mg Cd2+ l-1 for 21 days, the greatest Cd (n = 11; 9.99 +/- 5.09 micrograms g-1 wet mass) and Cu concentrations (n = 11; 17.90 +/- 4.66 micrograms g-1 wet mass) were associated with the gills, and to a lesser extent the digestive gland (n = 11; 0.38 +/- 0.20 microgram g-1 wet mass), whereas concentrations of Zn were variable in both organs. In the haemolymph Cd levels were relatively small (n = 11; 0.012-0.006 microgram ml-1) with exposure and time and Cu, Zn concentrations varied. Changes in the uptake of Cd in P. warreni indicated that transport, storage and possibly regulatory mechanisms are likely to operate in adult crabs. The potential of P. warreni as a bioindicator species of pollution is also discussed. PMID- 11383474 TI - 2,3,7,8-tetrachlorodibenzo-p-dioxin affects size and shape, but not asymmetry, of mandibles in mice. AB - Fluctuating asymmetry (FA), the random differences between the left and right sides of a bilaterally symmetrical character, is often purported to be a sensitive measure of developmental instability particularly in populations exposed to environmental stressors. As the level of developmental instability increases, often too does the level of FA. In this study we tested the hypothesis that exposure of pregnant mice to low doses of 2,3,7,8-tetrachlordibenzo-p-dioxin (TCDD) would increase the level of FA in the mandibles of their offspring. We used ten landmark coordinates around the mandible to create a single size variable (centroid size) and 20 Procrustes shape variables. These were used to test for effects of dioxin on mandible size and shape and their asymmetries. We found no detectable effect of TCDD on levels of FA in either size or shape of the mandible, but TCDD did produce a significant decrease in mandible size, and a significant effect on the overall shape. PMID- 11383475 TI - Why short-term bioassays are not meaningful--effects of a pesticide (Imidacloprid) and a metal (cadmium) on pea aphids (Acyrthosiphon pisum Harris). AB - The life-table study on effects of a heavy metal, Cd (100 and 200 mg kg-1 soil), and an insecticide, imidacloprid (4 and 40 g a.i. ha-1), on pea aphids revealed significant effects of both chemicals on life history traits and population dynamics. Substantial differences in the action of the two chemicals and between the two doses of imidacloprid were observed. The pesticide caused high mortality at the beginning of the experiment, however some aphids were able to live for as long as control insects. In contrast, Cd-induced mortality was low at the beginning but increased during the experiment. Analysis of data collected during the first 10 days of the experiment, the first 20 days, or for the whole life time of the cohort gave different results. While highly significant effects of imidacloprid were detected already after 10 days of the experiment, Cd treatment resulted in no effect for this time of exposure. Higher dose of imidacloprid decreased growth and development rates along with delayed reproduction (9th or 10th day). In contrast, the aphids treated with the low dose of imidacloprid and with both concentrations of Cd started their reproduction at the same day as untreated insects (6th or 7th day), but the reproduction tended to cease earlier. This indicates that too short ecotoxicological tests may result in serious underestimation of some effects, while overestimating others. PMID- 11383476 TI - Trace metal toxicity to phytoplankton of Kuwait coastal waters. AB - Trace metal concentrations in phytoplankton were determined in seven locations (Stations I-VII) of the Kuwait Coast, known for significant industrial and recreational activities. Trace metal concentrations in phytoplankton (20-200 microns size) occurred in the sequence of Zn > Pb > Cu > Ni at Stations I-III and VII, and Zn > Cu > Ni > Pb at Stations IV-VI. Station III showed higher concentrations of all the analyzed metals than at other stations. Toxicity tests to phytoplankton revealed a significant decrease in the rate of oxygen with increasing concentrations of trace metals. Further, such comparisons to station wise observations between the rate of oxygen evolution and trace metal levels revealed the nature of trace metal pollution in each station off the Kuwait coast. PMID- 11383477 TI - Localization of inhibin/activin subunits in normal pituitary and in pituitary adenomas. AB - The localization of inhibin/activin (I/A) subunits was investigated in human normal adenohypophysial cells and in 87 pituitary adenomas of different types, using immunohistochemistry. Monoclonal antibodies directed against alpha, beta A and beta B subunits of I/A were employed. In normal pituitary, alpha subunit of inhibin was detected only in FSH-positive gonadotrophs, while beta A subunit of I/A was expressed in FSH-positive gonadotrophs, GH-cells and in a few PRL-cells. beta B subunit was found in FSH-positive gonadotrophs, TSH-cells and a few LH positive gonadotrophs. The three subunits of I/A were detected in the majority of nonfunctioning tumors, while functioning adenomas showed a significantly lower expression. This study shows that alpha, beta A and beta B subunits of I/A are expressed by specific adenohypophysial cell types and that they are characteristically present in nonfunctioning adenomas. These results suggest that inhibins and activins may play a role in the local regulation of pituitary hormonal secretion both in normal adenohypophysial cells and in pituitary adenomas. PMID- 11383478 TI - Genes expressed in the mouse pituitary corticotrope AtT-20/D-16v tumor cell line. AB - The pituitary corticotrope AtT-20 stable cell line has been used as a model system to study peptide secretion, glucocorticoid regulation, and several other processes. In order to better understand this model cell line, a phage cDNA library was generated from AtT-20/D-16v cell mRNA and cDNA sequences were obtained for 317 clones representing 203 known genes and 48 novel cDNAs. The sequencing results revealed the prevalence of the mouse leukemia virus in this cell line and also identified a number of putatively secreted molecules that were not previously recognized as being secreted from AtT-20/D-16v cells or pituitary corticotropes. Nine completely novel cDNAs and 39 cDNAs homologous to known ESTs were also identified. A listing of other genes known to be expressed in AtT-20/D 16v cells is also provided. PMID- 11383479 TI - The in vitro inhibitory action of antiprogestin RU486 on LH and FSH secretion in the absence of progesterone in rats is estrogen-dependent. AB - Previous in vivo findings show that in the virtual absence of progesterone (P), the antiprogestin RU486 reduces LH and FSH secretion in proestrous rats, indicating that activation of P receptor (PR) can occur in the absence of the cognate ligand. The present study investigates, in vitro, whether or not the inhibitory effect of antiprogestin RU486 on gonadotropin secretion in the absence of P is estrous cycle dependent, and whether its specific expression in proestrus mirrors the high estrogen (E2) background. In the first experiment we investigated the effect of RU486 (10 nM) and/or LHRH (10 nM) on LH and FSH secretion in incubated pituitaries collected on each day of the estrous cycle of the rat. In the second experiment, we determined the effect of RU486 and/or LHRH on preovulatory LH and FSH release by pituitaries from female rats that were ovariectomized (OVX), treated with the antiestrogen LY117018-HCL (Eli Lilly & Co.), or injected with 20 micrograms of estradiol benzoate (EB). The third experiment investigated the effect of RU486 and/or LHRH on LH and FSH release by pituitaries collected from intact or EB-treated (0.1 mg/kg over three consecutive days) male rats. RU486 reduced both basal and LHRH-stimulated LH and FSH secretion in proestrous pituitaries from normal 4-day cyclic rats. By contrast, in diestrous pituitaries, RU486 increased both parameters of LH secretion but was without effect on FSH release. RU486 was also without effect in pituitaries collected from rats in estrus or metestrus, or from OVX or antiestrogen-treated rats. Moreover, EB injection or treatment induced the full inhibitory effect of RU486 in pituitaries from female and male rats, respectively. The above results suggested that P occupancy of the receptor is not required for the formation or function of the active receptor and hence for preovulatory LH and FSH secretion, and that this form of PR activation at pituitary level is E2-dependent and not genetically determined. PMID- 11383480 TI - Double pituitary lesions in three patients with Cushing's disease. AB - Double pituitary adenomas are rare in surgical specimens and the most common clinical feature in reported patients has been acromegaly. We report 3 cases of double pituitary lesions in patients who presented with Cushing's disease. In a 22-year-old man (case 1) with delayed puberty and low testosterone levels, mild hyperprolactinemia was diagnosed and treated with dopamine agonist therapy that reduced the prolactin (PRL) levels to normal. Over a 1-year period Cushing's disease developed gradually and was confirmed with classical endocrine testing. In a 27-year-old woman (case 2) who initially presented with severe depression and morbid obesity there was a gradual onset of Cushing's disease; initially she had minimally elevated serum PRL. In a 33-year-old woman (case 3) there was a 2 year history of Cushing's disease characterized by hirsutism, hypertension and weight gain; serum PRL was normal. Magnetic resonance imaging in all 3 patients revealed a microadenoma that was successfully removed by transsphenoidal pituitary surgery. Histology and immunocytochemistry in case 1 and case 3 revealed a corticotroph cell adenoma and a PRL cell adenoma in separate areas of the pituitary. In case 3 the PRL cell adenoma was "silent" but in case 1 the PRL cell adenoma may have been the cause of the mild hyperprolactinemia. In case 2 nodular corticotroph hyperplasia was the cause of Cushing's disease and a "silent" PRL cell adenoma was also identified. We conclude from these cases and a literature review that double pituitary lesions may occur in patients with Cushing's disease. The corticotroph part of the double lesion may consist of a corticotroph cell adenoma or, as reported in this study, of corticotroph nodular hyperplasia. The counterpart of the double lesion may consist either of a "silent" PRL cell adenoma or a functional PRL cell adenoma causing hyperprolactinemia. PMID- 11383481 TI - Glucagon is an ACTH secretagogue as effective as hCRH after intramuscolar administration while it is ineffective when given intravenously in normal subjects. AB - It is widely accepted that glucagon stimulates GH, ACTH and cortisol release in humans, though the mechanisms underlying these effects are unclear. Aim of the present study was to evaluate the stimulatory effect of intramuscolar (i.m.) and intravenous (i.v.) glucagon (GLU) administration on ACTH, cortisol (F) and GH release in normal adult subjects and to compare its effect on hypothalamo pituitary adrenal (HPA) axis with that of hCRH. To this goal, in 6 normal young women (26-32 yrs, 50-58 kg) we studied the ACTH and F responses to either i.m. or i.v. GLU (1 mg, approximately 0.017 mg/kg in subjects of 54.1 +/- 1.6 kg) administration as well as to i.v. hCRH (2.0 micrograms/kg) or placebo administration. The GH and glucose variations after GLU administration were also studied. I.v. GLU did not modify the spontaneous decrease of ACTH and cortisol levels observed after placebo. Conversely, i.m. GLU elicited clear-cut ACTH and F responses (peak vs baseline, mean +/- SEM: 53.0 +/- 15.2 vs 19.0 +/- 1.5 pg/ml, p < 0.05 and 222.3 +/- 23.8 vs 158.3 +/- 7.0 micrograms/l, p < 0.05) which were higher than those recorded after hCRH (28.1 +/- 4.6 vs 17.4 +/- 3.1 pg/ml, p < 0.02 and 182.7 +/- 22.8 vs 114.8 +/- 12.3 micrograms/l p < 0.02), though this difference did not attain statistical significance. Also GH rise was recorded after i.m. but not after i.v. GLU administration (11.6 +/- 3.4 vs 3.3 +/- 0.7 micrograms/l, p < 0.05). Thirty min after both i.v. and i.m. GLU administration glucose levels showed a similar increase followed by similar decrease. The intramuscular administration of GLU induced negligible side-effects in some subject (mild and transient nausea) which, on the contrary, were clear in all subjects after its intravenous administration (nausea, vomiting, tachycardia). In conclusion, glucagon "per se" is not an ACTH, cortisol and GH secretagogue. After intramuscular administration glucagon is a stimulus of HPA axis at least as effective as hCRH. The mechanisms underlying the ACTH, cortisol and GH responses to i.m. glucagon unlikely include glucose variations or stress. PMID- 11383482 TI - Growth hormone (GH) secretion in primary adrenal insufficiency: effects of cortisol withdrawal and patterned replacement on GH pulsatility and circadian rhythmicity. AB - We studied the effects of cortisol withdrawal and patterned replacement upon spontaneous GH secretion and circadian rhythmicity in 7 patients with Addison's disease. Hydrocortisone was administered in physiological daily total dosages, and all resulting plasma cortisol values were 2-15 micrograms/dl. It was given in 3 pulsatile modes: simulating "physiological" rhythm, "reverse" diurnal rhythmicity and "continuous" pulsatility. All modes of cortisol administration increased mean 24 h, GH pulse amplitude and interpulse GH levels. During saline infusions circadian GH rhythmicity was preserved, with GH being at its highest between 2400-0400 h. Administration of hydrocortisone in any mode did not modify circadian GH rhythmicity. We conclude: Cortisol replacement in physiological daily doses increases GH output in patients with Addison's disease by augmenting GH pulse amplitude and interpulse levels. This is likely due to the attenuation of hypothalamic somatostatin (SRIF) secretion by physiologic levels of cortisol. By inference, it implies that cortisol deficiency leads to diminution of GH output with low GH pulse amplitude, likely as a result of an augmented hypothalamic somatostatin secretion. However, circadian rhythmicity of GH secretion is glucocorticoid-independent. PMID- 11383483 TI - Hypopituitarism following closed head injury. AB - We describe four young patients (age 19-34 years) with hypopituitarism following closed head injury. The diagnosis was made by demonstration of low basal pituitary hormone levels and dynamic tests showing low pituitary reserve. The time interval between the injury and diagnosis of hypopituitarism was between three weeks and two months demonstrating the difficulty and complexity of making this diagnosis. Three of our patients (all patients suffering from anterior pituitary hormone deficiency) had ACTH deficiency, a condition which may be life threatening if left undiagnosed; these patients also demonstrated central hypothyroidism. Hypogonadotrophic hypogonadism occurred in three of the patients and was treated with hormonal replacement. Diabetes insipidus was the only insult in one of our patients, accompanied other hormonal deficits in two, and did not appear at all in another patient. Information about skull damage was available for three of the patients, and included skull base and facial bone fractures, probably reflecting the extent of injury necessary to cause hypopituitarism. All patients regained normal lives with adequate hormonal replacement therapy. PMID- 11383484 TI - Somatotropinoma infarction during octreotide therapy leading to bilateral cavernous sinus syndrome. AB - The cyclic somatostatin analog, octreotide, forms the mainstay of medical treatment for acromegaly. In addition to lowering circulating growth hormone levels and shrinking tumor size, octreotide may provide symptomatic relief of headaches associated with growth hormone secreting tumors. The majority of reported complications of octreotide therapy are gastrointestinal and metabolic. The present case illustrates the development of acute bilateral cavernous sinus syndrome with loss of eye movement bilaterally during octreotide therapy. Serial MRI examination suggest tumor infarction as the etiology. The symptoms resolved over 2 months as the tumor shrunk in size and growth hormone was dramatically reduced. PMID- 11383485 TI - Rapid re-expansion of a macroprolactinoma after early discontinuation of bromocriptine. AB - Prolactin (PRL)-secreting pituitary adenomas are the most common functioning pituitary tumors. Medical treatment with dopamine agonists is the therapy of choice for macroprolactinomas (> or = 10 mm). Withdrawal of bromocriptine after weeks or months of uninterrupted therapy has been associated with rapid tumor re expansion as evidenced by x-ray and CT scanning of the pituitary region. We report a patient with a giant macroprolactinoma who had a dramatic response to bromocriptine (tumor volume shrinkage of 53% within a month) but rapid re expansion to its original dimensions one week after discontinuation of bromocriptine. To our knowledge, this is the first time that the rapid shrinkage/re-expansion of a macroprolactinoma has been documented with serial MRI scans. PMID- 11383486 TI - Time course of GH and IGF-1 levels following withdrawal of long-acting octreotide in acromegaly. AB - AIM: Several studies have demonstrated the efficacy of octreotide LAR administered intramuscularly at 4-week intervals in the treatment of acromegaly. In contrast, few data are available on the time course of GH and IGF-1 plasma levels following octreotide LAR withdrawal. This prompted us to study these parameters for up to 20 weeks following drug withdrawal in a group of 18 acromegalic patients treated for one year. DESIGN AND PATIENTS: We studied 18 patients treated with octreotide LAR 10 mg (n = 2), 20 mg (n = 15) and 30 mg (n = 1) every 4 weeks for one year. GH (mean level during a 4-hour daily profile) and IGF-1 concentrations were measured at the end of treatment, just before the last injection (baseline) and then 15 +/- 2 weeks (first control) after the last injection. In patients with GH levels below 2.5 micrograms/L and/or normal IGF-1 at the first control, a second control was performed four to eight weeks later. RESULTS: After one year of treatment with octreotide LAR, the mean plasma GH concentration was 1.91 +/- 1.25 micrograms/L (mean +/- SE) and the mean IGF-1 concentration was 440 +/- 251 micrograms/L. Among the 18 patients, 13 had mean plasma GH concentrations below 2.5 micrograms/L and seven could be considered as well-controlled (normal IGF1 and mean GH levels below 2.5 micrograms/L). After treatment withdrawal, the plasma GH concentration remained below 2.5 micrograms/L at the first and the second controls in 2 of the 13 (15%) patients with suppressed GH levels on baseline. Among the seven well-controlled patients on baseline (GH levels below 2.5 micrograms/L and normal IGF-1), one (15%) remained well-controlled, one (15%) kept GH levels below 2.5 micrograms/L but increased IGF-1 levels, and one (15%) kept normal IGF-1 levels but increased mean GH levels at the first control. This hormonal status remained unchanged at the second control in these 3 patients. CONCLUSIONS: These results show long-lasting suppression of GH secretion after treatment withdrawal in some acromegalic patients treated for 12 months with octreotide LAR. The duration of GH suppression after treatment withdrawal is variable. Mean GH levels remained below 2.5 micrograms/L in 15% of our patients for up to 21 weeks following withdrawal of octreotide LAR. In practice, it may be preferable to wait several months after long-acting somatostatin analog withdrawal before reassessing hormone status. Owing this long-lasting effect, a dose reduction to 10 mg and/or a longer interval between injections could be considered for very good responders, as this would lead to considerable cost savings without affecting GH or IGF-1 control. PMID- 11383487 TI - Regioselective alkylation at the N4 position of a 3-oxo-1,4-benzodiazepine on solid support. AB - An efficient solid phase regioselective alkylation at the N4 position of a 3-oxo 1,4-benzodiazepine template exemplified by 4-H-2,3,4,5-tetrahydro-7-iodo-3-oxo-1H 1,4-benzodiazepine-2-acetate-polymer ester is described. Further chemical elaboration at position 7, utilizing a modified Heck reaction, allows the incorporation of amides from primary or secondary amines. The two diversity points at positions 4 and 7 were utilized to synthesize a 28-membered, combinatorial array on Sasrin resin in moderate yields and > 80% purity. Having validated the chemistry on solid support, a combine and split approach to prepare a bead-bound combinatorial library is achievable utilizing similar experimental practices and procedures as in the array synthesis. PMID- 11383488 TI - Comments on the design of chemical libraries for screening. AB - Different representations of molecules, based on distinct sets of properties can yield different perspectives of the issues involved in library design. In particular, different chemical representations can give rise to very different estimates of required library sizes. We provide a preliminary mathematical framework that examines the size of libraries required to adequately sample the spaces corresponding to some commonly used property sets. Introduction of conformational flexibility is also discussed as a means of increasing coverage of chemical libraries, while at the same time considering the thermodynamic consequences of flexibility upon detectable activity. Our theoretical analysis reveals that the property spaces currently in use are extremely large and unlikely to provide adequate discrimination among compounds. PMID- 11383489 TI - A safety-catch linker for amine release under biologically compatible conditions. AB - A 'biocompatible' safety-catch linker based on an internal diketopiperazine release has been developed to allow the release of amine based compounds. The synthesis of the linker and its pre-loading with a representative set of amines is described. After immobilization onto an amino Tentagel resin, the linker was activated and the amines were released under biological conditions at pH 8. PMID- 11383490 TI - Analysis of libraries encoded with GC tags: compound elution, tag decode analysis, and statistical sampling analysis. AB - Libraries encoded with electrophoric tags present a unique challenge with respect to library quality control and characterization. Libraries are prepared on Tentagel resin in 200-fold redundancy wherein each resin particle contains one compound per one tag set. The amount of compound present on the bead is ca. 200 500 pmole while tag levels are estimated at 0.5-1 pmol/bead. Several quality control protocols have been developed in order to accurately estimate bead yield and purity for the entire library, ensure high tag fidelity, and to determine the overall performance of individual synthons. This review provides a unique, collective portrait of Pharmacopeia's approach in assessing the quality of libraries prepared using its molecular encoding technology. PMID- 11383491 TI - Heteroduplex mobility assay (HMA) pre-screening: an improved strategy for the rapid identification of inserts selected from phage-displayed peptide libraries. AB - Phage-displayed peptide libraries represent an efficient tool to isolate peptides that bind a given target molecule. After several selection rounds, generally a large pool of target binding phages is obtained. Conventional analysis of the selected phage population involves extensive sequencing of many clones, most of which can be identical. We have adapted the Heteroduplex Mobility Assay (HMA) for pre-screening of phage inserts that were amplified by direct colony PCR of ELISA positive clones. This strategy allowed for the rapid and reproducible assignment of insert sequences to different 'heteroduplex migration groups'. Sequence analysis of only one representative of each HMA migration group then completes the characterisation of the binding phage population. In our model experiments, only 16% of HMA pre-screened clones required further sequence analysis. PMID- 11383492 TI - Preparing adult patients for international travel. AB - More than 50 million Americans engage in international travel annually. Travel to developing countries can expose patients to a myriad of infectious diseases. Primary care clinicians can provide counseling, immunizations, and drugs to keep patients healthy while traveling. This article reviews risk assessment and required and recommended immunizations. Patients can greatly reduce their risk for travel-acquired illness by updating immunizations, taking prophylactic drugs, and observing recommended health behaviors. PMID- 11383493 TI - Evaluating and managing osteoporosis in men. AB - Osteoporosis affects approximately 10 million Americans; of these, 2 million are men. An estimated 3.5 million additional men are at risk of developing the disease. Individuals with osteoporosis commonly incur fractures of the spine, hip, and forearm. The clinical spectrum of osteoporosis is similar in men and women; however, differences exist in skeletal development, age-related bone loss, modifiable and nonmodifiable risk factors, and secondary causes. Prevention and early detection is achieved through identification of risk factors and secondary causes. Treatment options include risk factor reduction, correction of underlying disease, and use of pharmacologic and nonpharmacologic therapies. PMID- 11383494 TI - A new conjugate vaccine against pneumococcal disease. AB - Streptococcus pneumoniae (pneumococcus) is a major cause of morbidity and mortality, particularly among infants and children. Pneumococcal 7-valent conjugate vaccine (PNCRM7) is the first conjugate vaccine known to prevent most invasive pneumococcal disease in infants and children. PNCRM7, which has a favorable safety profile, provides protection against invasive disease caused by antibiotic-resistant strains of S. pneumoniae, and the vaccine has demonstrated a significant impact on otitis media recurrence. Routine immunization with this vaccine should substantially reduce morbidity and mortality and improve the quality of life for infants, children, and their families. PMID- 11383495 TI - Considering evidence-based practice. PMID- 11383496 TI - Can business and science coexist in this century? PMID- 11383497 TI - A higher calling. Arkansas doctors lend a helping hand to Hondurans. PMID- 11383498 TI - Posterior dislocation of the shoulder is uncommon, hard to diagnose. PMID- 11383499 TI - Nasal meningioma: report of one case and review. AB - A case of primary nasal meningioma in a 69-year-old women is described. The pathologic, radiologic and clinical characteristics are described. A summary of previously published articles on the subject is given. PMID- 11383500 TI - Enhancing patient safety preoperative antibiotic prophylaxis. PMID- 11383501 TI - Common urologic problems in children: guides to evaluation and referral, Part I. AB - A discussion of common urologic problems in children is presented to provide primary physicians with appropriate guidelines for evaluation and referrals. The problems will be discussed in two parts: Part I will cover urinary tract infections, voiding dysfunctions, hematuria and proteinuria. Part II will cover abnormalities found on antenatal renal ultrasonography, hypospadias and other penile anomalies, phimosis, undescended testes, inguinal hernia and hydrocele, and varicoceles. An adage states: "The questions in medicine never change over time--only the answers." Certainly the busy primary care physician may experience the frustration of changing evaluation guidelines established by narrow subspecialties. Guidelines for the evaluation of children with disorders of the genitourinary tract are no exception. The following presentation will address some of the most common childhood urologic problems with a brief discussion of how to evaluate and when to refer for pediatric urologic consultation or management. PMID- 11383502 TI - Meningococcal disease, serogroup W135. PMID- 11383503 TI - Smallpox eradication: temporary retention of variola virus stocks. PMID- 11383504 TI - Proceedings of the symposium on chronic obstructive pulmonary disease: pathogenesis to treatment. London, United Kingdom, 25-27 January 2000. PMID- 11383505 TI - American Institute for Cancer Research 10th annual research conference: The role of nutrition in preventing and treating breast and prostate cancer. Washington, DC, USA. August 31-September 1, 2000. Abstracts. PMID- 11383506 TI - 114th AOAC international annual meeting reports and transactions. PMID- 11383507 TI - Biocompatibility of New Peritoneal Dialysis Solutions. Proceedings of a seminar. Seoul, Korea, 26 August 2000. PMID- 11383508 TI - Free Radicals in Life Science. Proceedings of a symposium. Tokyo, Japan, 9-10 March 2000. PMID- 11383509 TI - The role of quantum electro dynamics (QED) in medicine. Proceedings of a meeting. Rome, Italy, 14 December 1999. PMID- 11383510 TI - Heat shock protein 27 is a substrate of cGMP-dependent protein kinase in intact human platelets: phosphorylation-induced actin polymerization caused by HSP27 mutants. AB - Phosphorylation of heat shock protein 27 (Hsp27) in human platelets by mitogen activated protein kinase-activated protein kinase (MAPKAP) 2 is associated with signaling events involved in platelet aggregation and regulation of microfilament organization. We now show that Hsp27 is also phosphorylated by cGMP-dependent protein kinase (cGK), a signaling system important for the inhibition of platelet aggregation. Stimulation of washed platelets with 8-para-chlorophenylthio-cGMP, a cGK specific activator, resulted in a time-dependent phosphorylation of Hsp27. This is supported by the ability of cGK to phosphorylate Hsp27 in vitro to an extent comparable with the cGK-mediated phosphorylation of its established substrate vasodilator-stimulated phosphoprotein. Studies with Hsp27 mutants identified threonine 143 as a yet uncharacterized phosphorylation site in Hsp27 specifically targeted by cGK. To test the hypothesis that cGK could inhibit platelet aggregation by phosphorylating Hsp27 and interfering with the MAPKAP kinase phosphorylation of Hsp27, the known MAPKAP kinase 2-phosphorylation sites (Ser15, Ser78, and Ser82) as well as Thr143 were replaced by negatively charged amino acids, which are considered to mimic phosphate groups, and tested in actin polymerization experiments. Mimicry at the MAPKAP kinase 2 phosphorylation sites led to mutants with a stimulating effect on actin polymerization. Mutation of the cGK-specific site Thr143 alone had no effect on actin polymerization, but in the MAPKAP kinase 2 phosphorylation-mimicking mutant, this mutation reduced the stimulation of actin polymerization significantly. These data suggest that phosphorylation of Hsp27 and Hsp27-dependent regulation of actin microfilaments contribute to the inhibitory effects of cGK on platelet function. PMID- 11383511 TI - Yeast nuclear extract contains two major forms of RNA polymerase II mediator complexes. AB - The yeast Mediator complex is required for transcription by RNA polymerase II (pol II) in vivo and in vitro. This complex of over 20 polypeptides associates with pol II and is recruited to transcription complexes at promoters. Previous isolations of yeast Mediator-containing complexes in different laboratories have identified several distinct complexes. To identify the major forms of Mediator in yeast, Mediator was isolated from nuclear extracts using a two-step chromatographic procedure, avoiding ion exchange chromatography and high salt conditions to prevent dissociation of subunits during purification. Components of the Mediator complexes were identified by mass spectrometry and Western analysis. The major form of Mediator, termed pol II x Med, contained pol II and Mediator, including the Srb8-11 module. A second lower molecular size complex was also identified, termed Mediator core (Medc), which lacked pol II, Srb8-11, Rox3, Nut1, and the Rgrl module. Both of these complexes were active in transcription in vitro, although the Medc complex had significantly lower activity and could compete with the activity of the pol II x Med complex in vitro. PMID- 11383512 TI - Abstracts of the 15th European Histocompatibility Conference. March 27-30, 2001. Granada, Spain. PMID- 11383513 TI - Comment on 'Quantitative estimation of rare adverse events which follow a biological progression: a new model applied to chronic NSAID use' Tramer et al., Pain 2000;85:169-182. PMID- 11383514 TI - Comment on 'Differences in somatic perception in female patients with irritable bowel syndrome with and without fibromyalgia' l. Chang et al, PAIN 84 (2000) 297- 307. PMID- 11383515 TI - [Physiopathology and management of the aged--physiopathology and care of exudative otitis media of the aged]. PMID- 11383516 TI - Re: "Carbon monoxide poisoning presenting as an afebrile seizure.". PMID- 11383517 TI - Microarrays in disarray? PMID- 11383518 TI - Millennium boom. PMID- 11383519 TI - Green politics' bitter harvest. PMID- 11383520 TI - Web alert. PMID- 11383521 TI - Proceedings from the international symposium on occupational and environmental allergy and immune diseases. Chieti, Italy, April 27-30, 1999. PMID- 11383522 TI - [VII CPA Symposium: The quality of extracorporeal erythrocytes. Mainz, 1-2 October 1999. Proceedings]. PMID- 11383523 TI - Skin colour in digital images. PMID- 11383524 TI - Treatment of burns: new perspectives for highly purified tannic acid? PMID- 11383525 TI - Umbilical burns in infants: an unusual complication of topical silver nitrate therapy. PMID- 11383526 TI - Recent references. PMID- 11383527 TI - The effect of porous microstructure on the anisotropy of bone-like tissue: a counterexample. PMID- 11383528 TI - Proceedings of the joint meeting of the Israeli Societies of Rheumatology, and Allergy and Clinical Immunology. Dead Sea, 7-8 December 2000. PMID- 11383529 TI - International Conference on Complementary, Alternative, and Integrative Medicine Research. San Francisco, California, USA. May 17-19, 2001. Abstracts. PMID- 11383530 TI - Olive oil and colorectal cancer. PMID- 11383531 TI - Altered alveolar mechanics in the acutely injured lung. AB - OBJECTIVES: Alterations in alveolar mechanics (i.e., the dynamic change in alveolar size during tidal ventilation) are thought to play a critical role in acute lung injuries such as acute respiratory distress syndrome (ARDS). In this study, we describe and quantify the dynamic changes in alveolar mechanics of individual alveoli in a porcine ARDS model by direct visualization using in vivo microscopy. DESIGN: Prospective, observational, controlled study. SETTING: University research laboratory. SUBJECTS: Ten adult pigs. INTERVENTIONS: Pigs were anesthetized and placed on mechanical ventilation, underwent a left thoracotomy, and were separated into the following two groups post hoc: a control group of instrumented animals with no lung injury (n = 5), and a lung injury group in which lung injury was induced by tracheal Tween instillation, causing surfactant deactivation (n = 5). Pulmonary and systemic hemodynamics, blood gases, lung pressures, subpleural blood flow (laser Doppler), and alveolar mechanics (in vivo microscopy) were measured in both groups. Alveolar size was measured at peak inspiration (I) and end expiration (E) on individual subpleural alveoli by image analysis. Histologic sections of lung tissue were taken at necropsy from the injury group. MEASUREMENTS AND MAIN RESULTS: In the acutely injured lung, three distinct alveolar inflation-deflation patterns were observed and classified: type I alveoli (n = 37) changed size minimally (I - EDelta = 367 +/- 88 microm2) during tidal ventilation; type II alveoli (n = 37) changed size dramatically (I - EDelta = 9326 +/- 1010 microm2) with tidal ventilation but did not totally collapse at end expiration; and type III alveoli (n = 12) demonstrated an even greater size change than did type II alveoli (I - EDelta = 15,418 +/- 1995 microm2), and were distinguished from type II in that they totally collapsed at end expiration (atelectasis) and reinflated during inspiration. We have termed the abnormal alveolar inflation pattern of type II and III alveoli "repetitive alveolar collapse and expansion" (RACE). RACE describes all alveoli that visibly change volume with ventilation, regardless of whether these alveoli collapse totally (type III) at end expiration. Thus, the term "collapse" in RACE refers to a visibly obvious collapse of the alveolus during expiration, whether this collapse is total or partial. In the normal lung, all alveoli measured exhibited type I mechanics. Alveoli were significantly larger at peak inspiration in type II (18,266 +/- 1317 microm2, n = 37) and III (15,418 +/- 1995 microm2, n = 12) alveoli as compared with type I (8214 +/- 655 microm2, n = 37). Tween caused a heterogenous lung injury with areas of normal alveolar mechanics adjacent to areas of abnormal alveolar mechanics. Subsequent histologic sections from normal areas exhibited no pathology, whereas lung tissue from areas with RACE mechanics demonstrated alveolar collapse, atelectasis, and leukocyte infiltration. CONCLUSION: Alveolar mechanics are altered in the acutely injured lung as demonstrated by the development of alveolar instability (RACE) and the increase in alveolar size at peak inspiration. Alveolar instability varied from alveolus to alveolus in the same microscopic field and included alveoli that changed area greatly with tidal ventilation but remained patent at end expiration and those that totally collapsed and reexpanded with each breath. Thus, alterations in alveolar mechanics in the acutely injured lung are complex, and attempts to assess what may be occurring at the alveolar level from analysis of inflection points on the whole-lung pressure/volume curve are likely to be erroneous. We speculate that the mechanism of ventilator-induced lung injury may involve altered alveolar mechanics, specifically RACE and alveolar overdistension. PMID- 11383532 TI - The art of negotiating. PMID- 11383533 TI - Do variations in cardiac output influence lung edema? PMID- 11383534 TI - Concern about records of fibrosing colonopathy study. PMID- 11383535 TI - Hymenoptera stings and serum tryptase. PMID- 11383536 TI - Use of Shewart's technique. PMID- 11383537 TI - Use of Shewart's technique. PMID- 11383538 TI - Use of Shewart's technique. PMID- 11383539 TI - Duchenne muscular dystrophy or Meryon's disease. PMID- 11383540 TI - Suicide of the nephron is preventable. PMID- 11383541 TI - Treatment of uterine fibroids. PMID- 11383542 TI - Treatment of uterine fibroids. PMID- 11383543 TI - Management of peanut allergy. PMID- 11383544 TI - Management of peanut allergy. PMID- 11383545 TI - Risks from depleted uranium. PMID- 11383546 TI - Risks from depleted uranium. PMID- 11383547 TI - Health-seeking behaviour for cough. PMID- 11383548 TI - Glycosylation of transferrin in Alzheimer's disease and alcohol-induced dementia. AB - Transferrin is a glycosylated metal-binding serum protein. Carbohydrate-deficient transferrin (CDT) is a marker of recent and heavy alcohol intake. A genetic variant of transferrin, TfC2, occurs with increased frequency in patients with Alzheimer's disease (AD). Hence the question arose whether, in addition to an altered amino acid sequence, there could also be a difference in the glycosylation state of transferrin in patients with dementia. Serum samples of 37 AD and 13 Alcohol-induced dementia patients as well as 10 healthy controls were analyzed for abnormal Tf variants, using isoelectric focusing followed by blotting with anti-Tf antibodies. This allowed the direct visualization of glycosylation variants of transferrin, and assessment of any increase in underglycosylated forms (di-, mono- and asialo transferrin). PMID- 11383549 TI - An investigation into the neuroprotective properties of ibuprofen. AB - There is increasing evidence suggesting a protective role for anti-inflammatory medications in neurological disorders such as Alzheimer's disease (AD). While there has not been any direct evidence for this, a number of clinical studies indicate that those patients who have had a history of nonsteroidal anti inflammatory use, have a lower incidence of AD. Since there is currently no evidence on the mechanism by which these agents offer possible neuroprotection, we investigated the potential neuroprotective properties of the nonsteroidal anti inflammatory drug, ibuprofen, by examining whether this agent could reduce lipid peroxidation and superoxide radical generation. Quinolinic acid and cyanide, known neurotoxins, were used to induce lipid peroxidation and superoxide anion formation respectively, in rat brain homogenate. The results show that ibuprofen significantly (p<0.05) reduced quinolinic acid-induced lipid peroxidation and cyanide-induced superoxide production. The results of the present report therefore suggest a possible mechanism for the neuroprotective effect of ibuprofen. PMID- 11383550 TI - A comparative study of the effects of cholesterol, beta-sitosterol, beta sitosterol glucoside, dehydroepiandrosterone sulphate and melatonin on in vitro lipid peroxidation. AB - The free radical scavenging abilities of the structurally related steroids beta sitosterol, beta-sitosterol glucoside (plant sterols and sterolins), cholesterol, and dehydroepiandrosterone sulphate (DHEAS) were compared with melatonin (an efficient free radical scavenger) in an in vitro system which measures lipid peroxidation of platelet membranes in the presence of iron (Fe2+). Lipid peroxidation is a process whereby cellular membranes are damaged due to the oxidative deterioration of polyunsaturated lipids, which may lead to cell death and disease in living organisms. Substances such as vitamin E protect cellular membranes against oxidative damage due to their chemical structures. The steroids cholesterol, beta-sitosterol, beta-sitosterol glucoside and dehydroepiandrosterone (DHEA) are structurally related to each other. During aging, serum concentrations of DHEA, DHEAS and melatonin decrease, while the concentration of cholesterol tends to increase. The aim of the present study was to compare the role these substances play in lipid peroxidation over a wide concentration range. At concentrations lower than the free iron in the reaction mixture, all the steroids investigated decreased lipid peroxidation. At higher concentrations, cholesterol and beta-sitosterol increased lipid peroxidation, while DHEAS and melatonin continued to decrease lipid peroxidation. PMID- 11383551 TI - 17Beta-estradiol protects against quinolinic acid-induced lipid peroxidation in the rat brain. AB - The neurotoxin quinolinic acid has been identified as a causative agent in Huntington's disease and is a metabolite of the tryptophan pathway in the brain. In the present study, the in vivo and in vitro effect of 17beta-estradiol on lipid peroxidation induced by quinolinic acid was investigated. For the in vivo experiments ovariectomized female rats were administered with 100 microg 17beta estradiol daily for seven days prior to and seven days following the intrahippocampal injection of 1 micromol quinolinic acid. The level of lipid peroxidation in brain homogenate was investigated using the thiobarbituric acid test. The in vitro experiments were performed in brain homogenates of ovariectomized female rats. The homogenate was treated with quinolinic acid alone or in combination with 17beta-estradiol. Quinolinic acid increased lipid peroxidation in a dose dependent manner in vitro, while homogenate co-treated with 17beta-estradiol showed a significant reduction in lipid peroxidation. 17Beta-estradiol was also shown to be protective against quinolinic acid in vivo. These results could explain the neuroprotective effect of 17beta-estradiol. PMID- 11383552 TI - Early suppression of striatal cyclic GMP may predetermine the induction and severity of chronic haloperidol-induced vacous chewing movements. AB - Haloperidol persists in brain tissue long after discontinuation while haloperidol induced tardive dyskinesia often worsens after withdrawal of the drug. The mechanism of haloperidol-associated tardive dyskinesia is unknown, although neurotoxic pathways are suspected. Nitric oxide (NO) synthase (NOS) inhibitors exacerbate haloperidol-induced catalepsy, while haloperidol itself is a potent neuronal NOS inhibitor in vitro. Since NO and cGMP are involved in striatal neural plasticity, this study investigates a possible relation between cGMP and extrapyramidal symptoms as early predictors of haloperidol-associated tardive dyskinesia. Sprague-Dawley rats were administered either water or oral haloperidol (0.25 mg/kg/d p.o.) for 17 weeks, followed by 3 weeks withdrawal. Saline (i.p.) or the nNOS/guanylate cyclase inhibitor, methylene blue (5 mg/kg/d i.p.), were co-administered with haloperidol for the first three weeks of treatment. Vacous chewing movements (VCM's) were continuously monitored, followed by the determination of striatal cGMP and peripheral serum nitrogen oxide (NOx) levels. Chronic haloperidol engendered significant VCM's, with acute withdrawal associated with significantly reduced striatal cGMP levels as well as reduced serum NOx. Furthermore, suppressed cGMP levels were maintained and VCM's were significantly worse after early administration of methylene blue to the chronic haloperidol group. However, serum NOx was unchanged from control. We conclude that the central effects of chronic haloperidol on striatal NO-cGMP function persist for up to 3 weeks post-withdrawal. Moreover, suppression of striatal cGMP constitutes an early neuronal insult that determines the presence and intensity of haloperidol-associated motor dysfunction. PMID- 11383553 TI - Overcrowding induces anxiety and causes loss of serotonin 5HT-1a receptors in rats. AB - Unfavorable conditions under which children grow up may contribute significantly to the development of psychiatric abnormalities in adult life. We studied the effects of overcrowding at an early age and how it may result in anxiety later in life. Sprague-Dawley rats were housed 10 animals per cage, from birth until 4 weeks post-weaning. Rats housed 3 per cage served as controls. The Elevated Plus maze was used to determine their anxious behavior. Thereafter, the serotonergic system in the hippocampus was investigated. Overcrowded rats were significantly more anxious than controls. The number of 5HT-1a receptors in the hippocampus decreased significantly and their affinity for the ligand, OH-DPAT, increased significantly. There was no significant difference in the hippocampal levels of serotonin between overcrowded rats and controls. Our study shows that overcrowding during childhood can result in anxious behavior and that the serotonergic system appears to play a role in its manifestation. PMID- 11383554 TI - Increased glutamate-stimulated norepinephrine release from prefrontal cortex slices of spontaneously hypertensive rats. AB - Spontaneously hypertensive rats (SHR) have behavioral characteristics (hyperactivity, impulsiveness, poorly sustained attention) similar to the behavioral disturbances of children with attention-deficit hyperactivity disorder (ADHD). We have previously shown that dopaminergic and noradrenergic systems are disturbed in the prefrontal cortex of SHR compared to their normotensive Wistar Kyoto (WKY) control rats. It was of interest to determine whether the underlying neural circuits that use glutamate as a neurotransmitter function normally in the prefrontal cortex of SHR. An in vitro superfusion technique was used to demonstrate that glutamate caused a concentration-dependent stimulation of [3H]norepinephrine release from rat prefrontal cortex slices. Glutamate (100 microM and 1 mM) caused significantly greater release of norepinephrine from prefrontal cortex slices of SHR than from control slices. The effect of glutamate was not mediated by NMDA receptors, since NMDA (10 and 100 microM) did not exert any effect on norepinephrine release and MK-801 (10 microM) did not antagonize the effect of 100 microM glutamate. These results demonstrate that glutamate stimulates norepinephrine release from rat prefrontal cortex slices and that this increase is enhanced in SHR. The results are consistent with the suggestion that the noradrenergic system is overactive in prefrontal cortex of SHR, the animal model for ADHD. PMID- 11383555 TI - Development of a method to evaluate glutamate receptor function in rat barrel cortex slices. AB - The rat is a nocturnal animal and uses its vibrissae extensively to navigate its environment. The vibrissae are linked to a highly organized part of the sensory cortex, called the barrel cortex which contains spiny neurons that receive whisker specific thalamic input and distribute their output mainly within the cortical column. The aim of the present study was to develop a method to evaluate glutamate receptor function in the rat barrel cortex. Long Evans rats (90-160 g) were killed by cervical dislocation and decapitated. The brain was rapidly removed, cooled in a continuously oxygenated, ice-cold Hepes buffer (pH 7.4) and sliced using a vibratome to produce 0.35 mm slices. The barrel cortex was dissected from slices corresponding to 8.6 to 4.8 mm anterior to the interaural line and divided into rostral, middle and caudal regions. Depolarization-induced uptake of 45Ca2+ was achieved by incubating test slices in a high K+ (62.5 mM) buffer for 2 minutes at 35 degrees C. Potassium-stimulated uptake of 45Ca2+ into the rostral region was significantly lower than into middle and caudal regions of the barrel cortex. Glutamate had no effect. NMDA significantly increased uptake of 45Ca2+ into all regions of the barrel cortex. The technique is useful in determining NMDA receptor function and will be applied to study differences between spontaneously hypertensive rats (SHR) that are used as a model for attention deficit disorder and their normotensive control rats. PMID- 11383556 TI - Immunolocalization of plasma kallikrein in human brain. AB - Plasma kallikrein (PK) is a cofactor in blood coagulation and modulates inflammation through the release of bradykinin. Previously it was believed that plasma prekallikrein (PPK), the precursor of PK and a member of the serine protease superfamily, was synthesized exclusively by hepatocytes and secreted into circulation. However, recent studies show that the human brain contains a high level of PPK mRNA. In this study we sought to determine which areas of the brain express PK. Tissue from the spinal cord and 13 different regions of the human brain were collected at autopsy within 24h from death. Sections were probed using polyclonal antibodies (characterized by Western blotting) specific for PK. PK concentrations in extracts of these tissues were measured by ELISA. Immunolabeling of PK was observed in the cell bodies of the neurons of the hypothalamus, thalamus, spinal cord, cerebral cortex and brainstem. Positive PK immuno-reactivity was also demonstrated in the cytoplasm of the ependymal cells in sections of the hypothalamus and spinal cord. In addition, some fibre tracts of the pons, medulla and hippocampus as well as secretory cells of the pituitary gland also labeled. No immunoreactive PK was visualized in the choroid plexus or cerebellum. Our data demonstrate the cellular localization of PK in human brain. This work is supported by other studies that demonstrate PK mRNA in human heart, lung, trachea and brain. The cellular distribution of PK and kinin receptors in specific brain areas suggests a role for PK in the nervous system. PMID- 11383557 TI - Detection of tissue kallikrein and kinin B1 and B2 receptor mRNAs in human brain by in situ RT-PCR. AB - Tissue kallikrein (TK) and kinin receptors have been immuno-localized in various areas of the human nervous system, suggesting that the kallikrein-kinin system (KKS) may be functionally active in the brain. The aim of this study was to determine the cellular expression of TK and kinin B1 and B2 receptor mRNAs in specific regions of the human brain by in situ reverse transcriptase polymerase chain reaction. Autopsy samples of the brain, spinal cord, kidney and salivary gland were embedded in paraffin. Sections (5 microm), adhered onto silane coated glass slides, were treated with Proteinase K and DNase, followed by reverse transcription polymerase chain reaction with specific KKS primers and digoxigenin dUTP. Detection of the digoxigenin-label demonstrated localization of TK, B1 and B2 mRNAs in the cytoplasm of some neuronal cell bodies in the hypothalamus, thalamus, frontal cortex and spinal cord. TK mRNA was also observed in the ependymal cells lining the cerebral ventricles and epithelial cells of the choroid plexus. In the choroid plexus, only B1 gene expression was observed in some choroidal epithelial cells while no B2 labeling was detected. The identification of mRNAs to TK, B1 and B2 kinin receptors in human nervous tissue supports previous evidence for the presence of the KKS in the brain and confirms localized protein synthesis. PMID- 11383558 TI - Inborn errors of metabolism: a cause of abnormal brain development. AB - Brain malformations are caused by a disruption in the sequence of normal development by various environmental or genetic factors. By modifying the intrauterine milieu, inborn errors of metabolism may cause brain dysgenesis. However, this association is typically described in single case reports. The authors review the relationship between brain dysgenesis and specific inborn errors of metabolism. Peroxisomal disorders and fatty acid oxidation defects can produce migration defects. Pyruvate dehydrogenase deficiency, nonketotic hyperglycinemia, and maternal phenylketonuria preferentially cause a dysgenetic corpus callosum. Abnormal metabolism of folic acid causes neural tube defects, whereas defects in cholesterol metabolism may produce holoprosencephaly. Various mechanisms have been proposed to explain abnormal brain development in inborn errors of metabolism: production of a toxic or energy-deficient intrauterine milieu, modification of the content and function of membranes, or disturbance of the normal expression of intrauterine genes responsible for morphogenesis. The recognition of a metabolic disorder as the cause of the brain malformation has implications for both the care of the patient and for genetic counseling to prevent recurrence in subsequent pregnancies. PMID- 11383559 TI - Clinical utility of surface EMG: report of the Therapeutics and Technology Assessment Subcommittee of the American Academy of Neurology. PMID- 11383560 TI - Clinical utility of surface EMG: report of the Therapeutics and Technology Assessment Subcommittee of the American Academy of Neurology. PMID- 11383561 TI - Clinical utility of surface EMG: report of the Therapeutics and Technology Assessment Subcommittee of the American Academy of Neurology. PMID- 11383562 TI - Chronic urticaria in a 17-year-old patient with a past history of bowel disease. PMID- 11383563 TI - Morphological organ alterations and infectious diseases in brown trout Salmo trutta and rainbow trout Oncorhynchus mykiss exposed to polluted river water. AB - Poor water quality is discussed as a major factor causing a decline of brown trout populations in Swiss rivers. For our study we have chosen a river in the Swiss midlands, where the brown trout population has decreased dramatically during the last 10 yr and where feral fish have shown distinctive pathological alterations. The objective of our study was to investigate whether river water may be responsible for impaired fish health leading to an increased mortality in the river. In an active monitoring program, groups of brown and rainbow trout were exposed to polluted river water for 24 mo. Fish held in tap water served as a reference. Mortality, macroscopic and histopathologic changes, and infectious agents were investigated. Compared with the reference group, high mortality rates and severe pathological alterations of the inner organs were observed in fish held in river water. Especially gills, liver and kidney of these fish showed significantly higher changes than fish from tap water. These changes were dominated by degenerative and inflammatory reactions. Additionally, several infectious agents were diagnosed in fish exposed to river water. The most important findings were furunculosis and proliferative kidney disease. Brown trout seemed to be more sensitive than rainbow trout to environmental stress and infectious agents. PMID- 11383564 TI - Identification of infectious salmon anaemia virus in Atlantic salmon from Nova Scotia (Canada): evidence for functional strain differences. AB - Infectious salmon anaemia (ISA) is a serious disease responsible for high morbidity in farmed Atlantic salmon Salmo salar in Norway, Scotland and New Brunswick, Canada. Recent attempts to identify different strains of ISA virus (ISAV) based on nucleotide sequence variation have shown that the Norwegian and Scottish samples are similar to one another but markedly different from New Brunswick samples. These data may suggest the presence of different strains on each side of the Atlantic but no functional difference has been found with either strain. We describe the first identification and characterisation of ISAV in Atlantic salmon from Nova Scotia, Canada. Further, salmon infected with the Nova Scotia ISAV do not show typical ISAV pathology or mortality. Sequencing of this new strain showed it to possess greater similarity to ISAV from Norway and Scotland than to ISAV from New Brunswick. These findings are discussed in terms of a possible origin of the Nova Scotia ISAV strain and the existence of an avirulent ISAV strain. The impact of current strain variation studies on our knowledge of ISAV is also discussed. PMID- 11383565 TI - Genome sequence of a VP2/NS junction region of pillar cell necrosis virus (PCNV) in cultured Japanese eel Anguilla japonica. AB - Pillar cell necrosis virus (PCNV) is an aquatic birnavirus that was isolated from farmed Japanese eel experiencing mass mortality. In this study, a VP2/NS junction region in genome segment A of PCNV was amplified by reverse transcriptase polymerase chain reaction (RT-PCR) and sequenced. The VP2/NS region in PCNV had the highest homology with that of a strain Ab of infectious pancreatic necrosis virus (IPNV). This result revealed that PCNV belongs to birnavirus genogroup II. PMID- 11383566 TI - A salmonid cell line (TO) for production of infectious salmon anaemia virus (ISAV). AB - A new cell line designated TO which provides a high yield of infectious salmon anaemia virus (ISAV) has been established. The cells originate from head kidney leukocytes isolated from Atlantic salmon and grow well at 20 degrees C in EMEM with 5% CO2 and without CO2 supplement in HMEM. The cells have at present been passed more than 150 times and no changes in morphology, growth or virus production have been observed. The virus infection results in cytopathic effects (CPE) within 9 d, and the virus titre obtained from centrifuged and filtrated cell lysates, measured as TCID50, was about 10(9.1) ml(-1). The virus isolated from lysates of infected cells by a sucrose gradient provided purified ISAV when examined by silver stained SDS-PAGE. Salmon injected with diluted virus supernatant showed mortalities, hematocrit values and clinical signs in accordance with infectious salmon anaemia. PMID- 11383567 TI - A novel trypanoplasm-like flagellate Jarrellia atramenti n. g., n. sp. (Kinetoplastida: Bodonidae) and ciliates from the blowhole of a stranded pygmy sperm whale Kogia breviceps (Physeteridae): morphology, life cycle and potential pathogenicity. AB - The successful 6 mo rehabilitation of a stranded juvenile pygmy sperm whale Kogia breviceps afforded the opportunity to study the poorly known protozoan fauna of the upper respiratory tract of cetaceans. Mucus samples were collected by holding either a petri dish or glass slides over the blowhole for 3 to 5 exhalations; preparations were examined as wet mounts, and then stained with Wrights-Giemsa or Gram stain. Blood smears were stained with Wrights-Giemsa. Unidentified spindle shaped and unidentified broad ciliates, reported from the blowhole of the pygmy sperm whale for the first time, were seen only initially, while yeast-like organisms and bacteria were seen intermittently. Epithelial cells and white blood cells were often present in the blowhole mucus, but red blood cells were never seen. A novel trypanoplasm-like bodonid kinetoplastid biflagellate (Order Kinetoplastida) was commonly encountered in the blowhole mucus, but never in the blood. Both mature flagellates and those undergoing longitudinal binary fission were present. The elongate flagellate had a long whiplash anterior flagellum; the recurrent flagellum was attached along at least two-thirds of the body length, forming a prominent undulating membrane, and the trailing portion was short. The kinetoplast was irregularly fragmented. The flagellates were either free swimming, or attached to host material via the free portion of the posterior flagellum. The prominent undulating membrane was characteristic of Trypanoplasma, while the fragmented kinetoplast was characteristic of some species of Cryptobia. For the novel bodonid kinetoplastid, with its unique combination of morphological features (prominent undulating membrane and fragmented kinetoplast), we propose the creation of a new genus Jarrellia. We believe this to be the first published description of a flagellate from a marine mammal, and among the first reports of a trypanoplasm-like flagellate from a warm-blooded host. We expect that a diversity of flagellates and ciliates are commonly present in the blowhole of cetaceans. Future studies on the identity of the protozoans and the health of their cetacean hosts, which are readily studied in captivity, are necessary to establish their status as commensals or parasites. PMID- 11383568 TI - Characterization of intracytoplasmic prokaryote infections in Dreissena sp. (Bivalvia: Dreissenidae). AB - This study characterizes intracytoplasmic infections with prokaryote microorganisms in Dreissena sp. (near Dreissena polymorpha) from northeastern Greece and represents the first report of such infections in freshwater bivalves. Light microscope observations of stained tissues revealed basophilic, cytoplasmic inclusion bodies in 87.5% (28/32) of the mussels sectioned. Inclusions in epithelial cells and connective tissues were noted, respectively, in 34.4 and 71.9% of the sample, with 5 mussels (15.6%) having both tissue types infected. Epithelial cell infections were observed in histological sections only in digestive gland tubules and ducts; within tubules, inclusions were present more often in secretory than digestive cells. Connective tissue infections, however, were systemic; among the 32 mussels sectioned, inclusions were found in the gills (65.6%), foot (12.5%), mantle (9.4%), labial palps (6.3%), digestive gland (6.3%), stomach (6.3%), and gonads (3.1%). Cytoplasmic inclusions (maximum dimension, 138 microm) were prominent enough in the gills to be visible in 17.0% of the 247 mussels dissected. Ultrastructurally, prokaryote cells in gill connective tissues were clearly characteristic of Chlamydiales-like organisms, with each intracytoplasmic inclusion containing a loosely packed mixture of elementary, reticulate, intermediate bodies, and blebs. Prokaryote colonies in digestive gland epithelial cells exclusively contained 1 of 4 morphological cell types and were considered Rickettsiales-like. Hexagonal, virus-like particles were present in the cytoplasm of the largest of these Rickettsiales-like prokaryotes. Although host stress was evident from localized cell necrosis and dense hemocyte infiltration, overall infection was fairly benign, with no major, adverse impact on body condition evident among sectioned or dissected mussels. A possible negative effect was partial constriction of gill water tubes, but at the infection intensity observed (typical range 1 to 7 inclusion bodies per section), significant interference with respiration and other metabolic functions of the gills was highly unlikely. PMID- 11383569 TI - Occurrence of myxosporean parasites in the gills of two tilapia species from Lake Nokoue (Benin, West Africa): effect of host size and sex, and seasonal patterns of infection. AB - The gill myxosporean parasites of 2 euryhaline tilapias from Lake Nokoue, Sarotherodon melanotheron melanotheron (Ruppel, 1853) and Tilapia zillii (Gervais, 1852), were investigated from October 1987 to October 1989. A total of 391 S. m. melanotheron and 222 T. zillii were examined. Both of the fish species studied were infected by 3 host-specific myxosporean parasites for which prevalence greatly varied during our investigations. The 2 most common ones were Myxobolus sp. and M. zillii, which were located in the branchial filament. No significant fish sex effect was found for these 6 different myxosporean parasites. As seasonal pattern was clearly demonstrated for M. zillii while a host size effect was found for M. dossoui. However, further investigations of these myxosporean infections are necessary to determine the real effect of these parasites on their host, as host fecundity and survival was not assessed. PMID- 11383570 TI - Redescription of Microsporidium takedai (Awakura, 1974) as Kabatana takedai (Awakura, 1974) comb. n. AB - Ultrastructural study of the microsporidian Microsporidium takedai from the muscles of masu salmon Oncorhynchus masou proved that this species can be assigned to the genus Kabatana Lom, Dykova and Tonguthai, 2000. The parasites develop within disintegrated sarcoplasm without any delimiting boundary or cyst. Cylindrical multinucleate meronts proliferate by serial constrictions into uninucleate stages which repeat the process. Eventually, the uninucleate stages transform into uninucleate sporonts, which divide once to produce sporoblasts, thus functioning as sporoblast mother cells. Spores, with a subterminally located anchoring disc and 3 to 4 turns of the polar tube coil, average 3.3 by 1.9 microm in size. The exospore is divided into small fields; the endospore frequently makes small invaginations into the spore inside. Phylogenetic analysis using SSU rDNA sequence consistently placed Kabatana takedai in a group consisting of Microgemma sp., Spraguea lophii and Glugea americanus. The K. takedai could easily be separated from the other species in the same group by 2 inserts in the SSU rDNA sequence. PMID- 11383571 TI - Purification of Piscirickettsia salmonis and associated phage particles. AB - Piscirickettsia salmonis was isolated from cell culture using differential centrifugation and purified on a 30% Percol gradient. The purity of the preparation was assessed by transmission electron microscopy and phage-like particles were found to be associated with some of the P. salmonis isolates examined. This is believed to be the first report of a phage associated with rickettsia from fish. PMID- 11383572 TI - Electron microscopic evidence that expression of capsular polysaccharide by Photobacterium damselae subsp. piscicida is dependent on iron availability and growth phase. AB - The expression of capsular polysaccharide (CPS) by the fish pathogen Photobacterium damselae subsp. piscicida was analysed in the virulent strain DI 21 in relation to the growth phase and presence or absence of available iron in the culture medium. Bacterial cells were processed for electron microscopy by a procedure that improves visualisation of the capsule through stabilisation with polycationic ferritin, and electron micrographs of ultrathin sections were scanned with an acquired computerised image analyser to measure capsular area. Cells grown under iron-limited conditions always had a significantly lower amount of capsular material on their surfaces than iron-supplemented cells, even when cells from different culture phases were compared. Irrespective of the presence or absence of iron in the culture medium the amount of CPS decreased with the age of the culture, i.e., from early log phase to late log phase to stationary phase. The in vivo significance of this regulatory role of iron remains to be investigated. PMID- 11383573 TI - Factors determining admission to neonatal units in Jamaica. AB - In order to identify the factors associated with admission to neonatal care units in a developing country, 1,823 newborns admitted to Jamaica's eight neonatal care units over a 6-month period were compared with 9,563 newborns identified during an island-wide population morbidity study. Maternal sociodemographic characteristics, past obstetric history, infant's growth parameters at birth and mode and place of delivery were investigated. Babies of mothers resident in the two regions of the island where specialist paediatric services were available had increased odds of admission (OR= 1.45, 1.22) compared with those living elsewhere (OR=0.70, 0.80). Maternal history of a previous miscarriage, termination or early neonatal death were associated with subsequent admission, but a previous stillbirth or late neonatal death were not. Very low birthweight infants of gestational age 28-31 weeks were more likely to be admitted than those < 28 weeks with ORs of 1.45 and 0.34 respectively. Factors determining neonatal admission in the developing world may be quite different from those of developed countries. The development of guidelines and support services to ensure wider access to these services for those most in need could contribute to more equitable utilisation of services. PMID- 11383574 TI - Role of structural birth defects in preterm delivery. AB - The proportion of preterm births associated with structural birth defects has not been adequately quantified. We explored the proportion of preterm infants with structural birth defects diagnosed in the first year of life, as well as the specific phenotypes of birth defects occurring among preterm infants. The data used were from the California Birth Defects Monitoring Program, a population based registry, as well as data from California vital records corresponding to births and fetal deaths in the period 1984-96. The prevalence of structural birth defects exceeded 8% among deliveries with gestational ages < or = 30 weeks, and prevalence decreased to 2% as gestational age increased to > or = 37 weeks gestation. The decreasing prevalence pattern with increasing gestational age was observed for a variety of anatomically defined birth defect groups suggesting that certain birth defects were not the sole contributors to the elevated prevalences among preterm births. Decreasing prevalence with increasing gestational age was also observed across strata of maternal race/ethnicities, ages, infant's sex and each year studied. These data indicate that structural birth defects may contribute significantly to the proportion of infants who are delivered before 37 weeks gestation. PMID- 11383575 TI - Testing a sociomedical model for preterm delivery. AB - The study purpose was to contribute to the development of a model for preterm birth that brings together social, psychosocial and biomedical factors. The three research questions were: (a) Which social and psychosocial factors influence the risk of preterm delivery? (b) Are the effects of social and psychosocial factors independent? (c) Do the biomedical factors identified and measured here explain the effect of social and psychosocial factors? The sample comprised 739 low income black non-Hispanic women. Interviews were conducted after delivery, and medical records were abstracted. Nearly a quarter of the women delivered preterm (23.5%). Two social factors strongly predicted risk of preterm delivery: inadequacy of time and money for non-essentials. The effect of both was stronger than expected from each alone and was partially mediated by psychosocial factors. Several psychosocial factors also predicted preterm delivery risk but only stress and locus of control were independent predictors. With the inclusion of biomedical factors, stress (OR = 1.86, P = 0.005) and locus of control (OR = 1.75, P = 0.007) continued to be strongly associated with preterm delivery. The effect of inadequate resources for non-essentials was no longer significant, suggesting mediation. These strong effects of social and psychosocial factors should be examined in future studies. PMID- 11383576 TI - Risk factors for spontaneous preterm birth in two urban areas of Ukraine. AB - The economic, social and health problems faced by former eastern bloc countries after the demise of the Soviet Union are unique in the recent history of Europe. We conducted a study in two urban areas of Ukraine, asking if the traditional predictors of preterm delivery continue to be associated with risk under these conditions. Subjects were pregnant women with last menstrual period (LMP) between 25 December 1992 and 23 July 1994. Self-completed questionnaires and the medical record provided data. We compared 137 spontaneous preterm deliveries with 2,886 full-term births, using all established risk factors for which we had data. Maternal age was the variable most strongly related to preterm birth. Being 18 or less had an odds ratio (OR) of 3.7; being 30+ had an OR of 2.5 relative to the reference group of age 25-29. Placental complications and pre-existing hypertension had ORs of 2.7 and 2.3, respectively, but the confidence interval included 1.0. Low net pregnancy weight gain (less than 10 kg) was significantly associated with preterm birth, but the rate of net weight gain was not. Marital status and educational category were only weakly related. We conclude that although Ukraine faced serious difficulties during its transition to a market economy, these problems did not generally alter the outcome of pregnancy in our sample when the classic risk factors for preterm delivery were present. PMID- 11383577 TI - Risks of induction of labour in uncomplicated term pregnancies. AB - Our objective was to evaluate the risks of maternal and perinatal morbidity associated with induction of labour in uncomplicated term pregnancies. We conducted a retrospective cohort study including 7,430 women, not referred from another institution, with a single baby in vertex presentation, and delivering between 38 and 40 weeks of pregnancy. Among these women, 3,546 were excluded for prelabour pregnancy complications. Relative risks (RR), adjusted for parity, were computed to compare 3,353 women who went into labour spontaneously with 531 women whose labour was induced. Induction of labour was found to be associated with a higher risk of caesarean section [RR = 2.4, 95% CI 1.8, 3.4]. Use of non-epidural [RR = 1.5, 95% CI 1.2, 1.8] and of epidural analgesia [RR = 1.4, 95% CI 1.1, 1.7] was more frequent after labour induction. Resuscitation [RR = 1.2, 95% CI 1.0, 1.5], admission to the intensive care unit [RR = 1.6, 95% CI 1.0, 2.4] and phototherapy [RR = 1.3, 95% CI 1.0, 1.6] were more frequent after induction of labour. Results were similar when controlling simultaneously for parity, maternal age, gestational age, year of delivery, birthweight and the physician in charge of delivery in a logistic regression analysis. The results of this study suggests that induction of labour is associated with a higher risk of caesarean section and of some perinatal adverse outcomes. Induction of labour should be reserved for cases where maternal and perinatal benefits outweigh the risk of these complications. PMID- 11383578 TI - Elective induction of labour: complicating the uncomplicated? PMID- 11383579 TI - Accuracy of self-reported cigarette smoking among pregnant women in the 1990s. AB - In large, prospective studies of pregnancy conducted in the 1960s, women reported very accurately whether or not they smoked. However, in the 1990s, pregnant women who smoke are often pressured to reduce or quit smoking, and the incentive to misreport may be greater than in the past. To assess the accuracy of reported smoking, the authors compared self-reported smoking with cotinine in the serum and/or urine of 105 women who participated in the Calcium for Pre-eclampsia Prevention pilot study in 1992. Cotinine confirmed the report of 84.6% of women who reported smoking and 94.5% of women who denied smoking. These fractions are virtually identical to those obtained in a pregnancy cohort from the 1960s. The authors conclude that in the setting of two obstetrical research studies not specifically focused on smoking, the accuracy of self-reported cigarette smoking did not change substantially from the 1960s to the 1990s. PMID- 11383580 TI - Review of risk factors for sudden infant death syndrome. AB - Sudden infant death syndrome (SIDS) accounts for the largest number of deaths during the first year of life in developed countries. The possible causes of SIDS are numerous and, to date, there is no adequate unifying pathological explanation for SIDS. Epidemiological studies have played a key role in identifying risk factors, knowledge of which has underpinned successful preventive programmes. This review critically assesses information on the main risk factors and causal hypotheses put forward for SIDS, focusing on research published since 1994. The overall picture that emerges from this review is that affected infants are not completely normal in development, but possess some inherent weakness, which may only become obvious when the infant is subjected to stress. Initially there may be some minor impairment or delay in development of respiratory, cardiovascular or neuromuscular function. None of these is likely to be sufficient, in isolation, to cause death and, provided the infant survives the first year of life, may no longer be of any significance. However, when a compromised infant is confronted with one or more stressful situations, several of which are now clearly identified as risk factors, and from which the majority of infants would normally escape, the combination may prove fatal. PMID- 11383581 TI - Neurosurgical exorcism. PMID- 11383582 TI - The incidence of lymphoid and myeloid malignancies among hospitalized Crohn's disease patients. AB - An association may exist between Crohn's disease (CD) and lymphoid/myeloid malignancies. We aimed to evaluate the 2-year cumulative incidence rate of lymphoid/myeloid malignancy among hospitalized CD patients. This is a retrospective cohort study using hospital discharge data from California and Virginia. Cohorts were defined by the presence or absence of a CD diagnosis in all patients discharged during a single calendar year (Year-2). The presence or absence of lymphoid/myeloid malignancy was determined for all hospitalizations during a 4-year period (Year-1 to Year-4) for each member of both cohorts. To obtain a 2-year cumulative incidence rate, patients with lymphoid/myeloid malignancy prior to or at the time of their first admission in Year-2 were excluded. Patients were followed for 8 quarters after this admission for the incidence of lymphoid/myeloid malignancy. Cumulative incidence rates and odds ratios were calculated. The crude 2-year incidence rate of lymphoid/myeloid malignancy among hospitalized CD patients was 3.87/1.000 CD patients (21/5,426; 95% CI = 2.40-5.92). The odds ratio adjusted for age, gender, and race was 2.04 (95% CI = 1.33-3.14, p < 0.001). The 2-year cumulative incidence of lymphoid/myeloid malignancies among hospitalized CD patients is greater than that seen in hospitalized patients without CD. This finding supports the need for further prospective population-based studies. PMID- 11383583 TI - Crohn's disease of the esophagus: clinical features and outcomes. AB - OBJECTIVES: Crohn's disease of the esophagus is rare. We sought to determine the clinical features and outcome of patients with esophageal Crohn's disease seen at our institution. METHODS: Patients with esophageal Crohn's disease evaluated at Mayo Clinic Rochester between 1976 and 1998 were identified. RESULTS: Twenty patients (0.2%) with esophageal involvement were identified. Median age at diagnosis was 31 years (range, 7-77 years). Eleven patients (55%) were female. Extraesophageal Crohn's disease preceded or was found at the same time as the diagnosis of esophageal Crohn's in all cases. Sixteen patients (80%) had symptoms referable to the esophagus. Endoscopic findings included ulcers in 17 (85%), erythema or erosions in 8 (40%), and strictures in 4 patients (20%). One patient had a fistula. The most common histological findings were active chronic inflammation (75%) and ulcer (30%). No granulomata were identified. Approximately one-half of our patients improved with first-line therapy. Eleven patients (55%) received immune modifier therapy. Six showed significant improvement on azathioprine, 6-mercaptopurine, or cyclosporine. Esophageal dilatation was required in six patients, and three patients required surgery. CONCLUSION: Esophageal Crohn's disease may be underdiagnosed. Patients with Crohn's disease complaining of esophageal symptoms should undergo upper endoscopy with biopsies, and the diagnosis of esophageal Crohn's disease should be entertained if aphthous or deep ulcers or strictures are present. Immune modifier therapy should be considered for steroid-dependent and steroid-resistant cases. PMID- 11383584 TI - Expression of inducible and endothelial nitric oxide synthases in pouchitis. AB - To study the induction of nitric oxide synthase (NOS) in different forms of pouchitis, we divided patients in five groups: 1) ulcerative colitis, no pouch; 2) no-pouchitis; 3) chronic asymptomatic pouchitis; 4) chronic active pouchitis; and 5) acute pouchitis. Ileal biopsies were scored for NOS-2 (inducible) and NOS 3 (endothelial) immunoreactivity and acute inflammation. In group 1, most specimens lacked NOS-2 immunoreactivity. In group 2, some specimens showed NOS-2 immunoreactive epithelium. In group 3, areas of NOS-2-immunoreactive epithelium were consistently observed in most specimens. In groups 4 and 5, most specimens showed moderate-to-extensive epithelial NOS-2 staining. NOS-2 immunoreactivity scores of groups 1-5 were 0.25 +/- 0.16, 0.67 +/- 0.19, 1.19 +/- 0.40, 2.0 +/- 0.23, and 2.18 +/- 0.12, respectively. Corresponding acute inflammation scores were 0, 0.53 + 0.17, 1.00 +/- 0.33, 1.80 +/- 0.20, and 1.64 +/- 0.15. NOS-2 score correlated with acute inflammation score (p < 0.0001), indicating that NOS-2 induction correlates with both the clinical degree of pouchitis and the severity of acute inflammation. NOS-3 immunoreactivity increased in all pouchitis groups. PMID- 11383585 TI - Inflammatory bowel disease-like colitis in glycogen storage disease type 1b. AB - Chronic inflammatory bowel disease (IBD)-like colitis is occasionally associated with glycogen storage disease-type 1b (GSD-1b). We describe a 17-year old boy with GSD-1b who developed an IBD-like colitis. Roentgenography and colonoscopy showed the lead-pipe appearance of the colon and circumferential ulcers. Histopathologic examination revealed nonspecific inflammation without granulomatous lesions. High-dose granulocyte-colony stimulating factor (G-CSF) and sulfasalazine led to the resolution of the colitis, although neutropenia continued. Besides this case, 10 published cases of GSD-1b and IBD-like colitis were reviewed. All cases had severe neutropenia and/or neutrophil dysfunction. The mean onset of bowel disease was 12.3 years of age. Seven cases required surgical treatment. All five patients with G-CSF/GM-CSF therapy showed clinical remission. These findings suggest that IBD-like colitis is a grave complication of GSD-1b and that recurrent enteric infections due to neutrophil deficiency may contribute to the development of this bowel disease. PMID- 11383586 TI - An investigation of the association of the prothrombin G20210A gene mutation and inflammatory bowel disease: Factor II and IBD. AB - BACKGROUND: A thrombotic etiology for inflammatory bowel disease (IBD) has been proposed as a result of its association with thromboembolic complications, smoking, the oral contraceptive pill, and the response of ulcerative colitis (UC) patients to heparin. We have previously demonstrated an increased prevalence of the Factor V Leiden mutation in UC and wished to investigate the frequency of the recently discovered prothrombin G20210A gene mutation in IBD. The aim of the study was to investigate the hypothesis that the prothrombic state associated with the prothrombin G20210A gene mutation is involved in the etiology of IBD. PATIENTS AND METHODS: A prospective cohort study of patients attending the Bristol Royal Infirmary and Gloucestershire Royal Hospital's IBD clinics was performed. Thirty-nine patients with IBD (24 with Crohn's disease and 15 with UC) and 100 historical controls were screened for the presence of the prothrombin gene mutation using a heteroduplex-based polymerase chain reaction technique. None of the patients with IBD had a personal history of thromboembolism, while three of them had a family history. RESULTS: No IBD patients had the prothrombin gene mutation compared with four (4%) controls (allelic frequency 2%). CONCLUSION: There does not appear to be an association of the prothrombin gene mutation with IBD and therefore it is unlikely to be involved in the etiology of IBD. PMID- 11383587 TI - Adaptation of bacteria to the intestinal niche: probiotics and gut disorder. AB - The gastrointestinal tract is a complex ecosystem host to a diverse and highly evolved microbial community composed of hundreds of different microbial species. The interactions that occur between this complex microbial community and the human host have become the focus of scientific research due to increases in the incidence of illnesses associated with deficient or compromised microflora (e.g., gastrointestinal tract infections, inflammatory bowel disease (Crohn's disease and ulcerative colitis), irritable bowel syndrome, antibiotic-induced diarrhea, constipation, food allergies, cardiovascular disease, and certain cancers). Effective multidisciplinary research programs now complement conventional microbiology with molecular ecology techniques to provide culture-independent analysis of the gastrointestinal ecosystem. Furthermore, as we acquire an understanding of gut microflora composition and processes such as intestinal adherence, colonization, translocation, and immunomodulation, we are also elucidating mechanisms by which these can be influenced. This knowledge not only allows scientists to define the activities and interactions of "functional food" borne beneficial bacteria in the gut, but will also provide the scientific basis for the development of innovative biotechnology-based products tailored to prevent specific diseases and promote overall human gastrointestinal health. PMID- 11383588 TI - Inflammatory bowel disease and pregnancy. AB - Most women with inflammatory bowel disease who desire to become pregnant can expect to conceive successfully, carry to term, and deliver a healthy infant. However, the management of inflammatory bowel disease during pregnancy remains challenging, and some women with ulcerative colitis or Crohn's disease will have difficulty becoming pregnant or have increased disease symptoms while pregnant. Control of disease activity before conception and during pregnancy is critical to optimize both maternal and fetal health. The natural history of inflammatory bowel disease during pregnancy will be reviewed and the medical and surgical therapy discussed. PMID- 11383589 TI - All patients with inflammatory bowel disease should have bone density assessment: pro. AB - Bone loss is a common problem for individuals with inflammatory bowel disease. Corticosteroids play an important role in the development of osteoporosis in these patients; however, active disease and longer disease duration also appear to increase the risk of osteoporosis. Given the current burden of osteoporosis, bone mineral density is recommended. PMID- 11383590 TI - All patients with inflammatory bowel disease should have bone density assessment: con. PMID- 11383591 TI - The importance of familial clusterings in Crohn's disease. PMID- 11383592 TI - Bones and Crohn's: should we treat Crohn's disease patients with alendronate? PMID- 11383593 TI - Growth of an IBD controversy: growth hormone and Crohn's disease. PMID- 11383594 TI - How safe is immunomodulator therapy? PMID- 11383595 TI - Infliximab in the treatment of severe, steroid-refractory ulcerative colitis: a pilot study. AB - We report the experience of 11 patients (of 60 planned patients) enrolled in a double-blind, placebo-controlled clinical trial of infliximab in patients with severe, active steroid-refractory ulcerative colitis. The study was terminated prematurely because of slow enrollment. Patients having active disease for at least 2 weeks and receiving at least 5 days of intravenous corticosteroids were eligible to receive a single intravenous infusion of infliximab at 5, 10, or 20 mg/kg body weight. The primary endpoint used in this study was treatment failure at 2 weeks after infusion. Treatment failure was defined as 1) unachieved clinical response as defined by a modified Truelove and Witts severity score, 2) increase in corticosteroid dosage, 3) addition of immunosuppressants, 4) colectomy, or 5) death. Safety evaluations included physical examination, clinical chemistry and hematology laboratory tests, and occurrence of adverse experiences. Four of 8 patients (50%) who received infliximab were considered treatment successes at 2 weeks, compared with none of 3 patients who received placebo. Improvement in erythrocyte sedimentation rates and serum concentrations of C-reactive protein and interleukin-6 correlated with the clinical response observed in patients receiving infliximab. Infusion with infliximab produced no significant adverse events. Infliximab was well tolerated and may provide clinical benefit for some patients with steroid-refractory ulcerative colitis. PMID- 11383596 TI - Is infliximab effective for induction of remission in patients with ulcerative colitis? PMID- 11383597 TI - Zinc supplementation tightens "leaky gut" in Crohn's disease. AB - OBJECTIVES: Small intestinal permeability is often increased in patients with Crohn's disease and may be pathogenic for clinical relapses. No effective prophylactic treatment is available for these patients. The aim of this study was to ascertain whether zinc supplementation may improve intestinal permeability. METHODS: We studied 12 patients with quiescent Crohn's disease who had been in remission for at least 3 months and had increased intestinal permeability on two separate occasions within the last 2 months. Patients received oral zinc sulfate supplements (110 mg three times a day) for 8 weeks and were followed-up for 12 months thereafter to monitor relapses. RESULTS: We found that the lactulose/mannitol ratio was significantly higher before supplementation than after (0.041 +/- 0.003 versus 0.026 +/- 0.005). During follow-up, 10 patients had normal intestinal permeability and did not relapse; of the remaining two who had increased intestinal permeability, one relapsed. CONCLUSIONS: Our findings show that zinc supplementation can resolve permeability alterations in patients with Crohn's disease in remission. Improving intestinal barrier function may contribute to reduce the risk of relapse in Crohn's disease. PMID- 11383599 TI - Multifunctional asymmetric catalysis. AB - Two types of general and practical enantioselective catalysts, namely, bimetallic complexes and Lewis acid-Lewis base bifunctional catalysts were developed based on the concept of multifunctional catalysis. In the first part of this review, the first example of a catalytic enatioselective nitro-Mannich reaction as well as a direct catalytic enantioselective aldol reaction of 2-hydroxyacetophenone using bimetallic complexes is discussed. The new complex, composed of ytterbium, potassium, and BINOL in a ratio of 1:1:3, promoted the nitro-Mannich reaction of nitromethane with up to 91% ee. On the other hand, second generation ALB catalyzed an enantioselective and diastereoselective nitro-Mannich reaction of nitroalkanes in up to 83% ee with a diastereomeric ratio up to 7:1. Moreover, the reaction of aldehydes with 2-hydroxyacetophenone in the presence of LLB, KHMDS, and H2O selectively gave the corresponding anti-alpha,beta-dihydroxy ketones in up to 95% ee and, in the presence of the catalyst prepared from linked-BINOL and 2 eq of Et2Zn, selectively afforded the syn-alpha,beta-dihydroxy ketones in up to 86% ee. In the second part, the development of new catalysts displaying a Lewis acidity and a Lewis basicity is described. The Lewis acid of the catalyst activates aldehydes, imines, acyl quinoliniums, and ketones. At the same time, the Lewis base activates the nucleophile (TMSCN). Catalysts of this type produced a highly enantioselective cyanation of these electrophiles. Application of the catalytic enantioselective cyanosilylation of aldehydes to a total synthesis of epothilones is also described. PMID- 11383598 TI - Impact of different therapeutic regimens on the outcome of patients with Crohn's disease of the upper gastrointestinal tract. AB - BACKGROUND: To follow-up clinical and biochemical features in patients with Crohn's disease (CD) of the upper gastrointestinal (GI) tract and to evaluate the impact of different therapeutics on the outcome of these patients. METHODS: 32 CD patients with endoscopically and histologically proven CD of the upper GI tract were included into this retrospective study. Gastroduodenal and intestinal permeability tests, inflammatory parameters, Crohn's Disease Activity Index (CDAI), and upper gastrointestinal complaint profile were sequentially assessed. These parameters were assessed at the beginning and followed up during therapies with antisecretory drugs, mesalamine, prednisolone, and azathioprine. RESULTS: Symptoms responded to antisecretory drugs. Gastroduodenal permeability increased under mesalamine. Gastroduodenal and intestinal permeability as well as CDAI decreased under prednisolone. Under azathioprine, gastroduodenal and intestinal permeability, CDAI, and C-reactive protein decreased. PMID- 11383600 TI - Evaluation of new pregnane derivatives as 5alpha-reductase inhibitor. AB - The objective of this study was to synthesize several new pregnane derivatives and evaluate them as antiandrogens. From the commercially available 16 dehydropregnenolone acetate (7), two new steroidal compounds were synthesized: 17alpha-hydroxy-17beta-methyl-16beta-phenyl-D-homoandrosta-1,4,6-triene-3,20 dione (18) and 17alpha-acetoxy-17beta-methyl-16beta-phenyl-D-homoandrosta-1,4,6 triene-3,20-dione (19). The 5alpha-reductase inhibitory effect of the new compounds 18 and 19 together with the previously synthesized intermediates 7, 8, 13, 16, and 17 was determined in three different models: gonadectomized hamster flank organs diameter size, incorporation of [1,2-(14)C]sodium acetate into lipids in flank organs and conversion of [3H]testosterone (T) to [3H]dihydrotestosterone (DHT) by Penicillium crustosum. The evaluation of these steroids was carried out with three different controls: one group was treated with vehicle, the second with T and the third group with T plus finasteride. The pharmacological results from this work demonstrated that T significantly increases the diameter of the pigmented spot on the flank organs (p<0.05) as well as the incorporation of labeled sodium acetate into lipids in gonadectomized hamster flank organs (from 0.125 to 0.255 nmol per gland). In this study we also observed that broth of Penicillium crustosum converted [3H]T to [3H]DHT in a manner comparable to that of the flank organs. All experiments indicated that finasteride as well as steroids 7, 8, 13, 16-19 reduced significantly the conversion of T to DHT in P. crustosum. These compounds also decrease the size of the pigmented spot in the flank organs as well as reducing the incorporation of radiolabeled sodium acetate into lipids; T and the control sample (treated with vehicle only) were used for comparison. Apparently the presence of the 4,6-diene 3,20-dione moiety and also the C-17 ester group produce a higher inhibitory effect on the parameters used. PPThe data from this study indicated also that the three models used for the pharmacological evaluation exhibited comparable results. PMID- 11383601 TI - Chemical conversion of cyclic alpha-amino acids to cyclic alpha-aminophosphonic acids. AB - The oxidative decarboxylation of cyclic alpha-amino acids having urethane-type N protecting groups with lead tetraacetate [Pb(OAc)4] gave 2-hydroxy derivatives, which were transformed into the corresponding alpha-aminophosphonic acid esters by treatment of trialkyl phosphites in the presence of Lewis acids. Deprotection and ester cleavage of the products in the usual manner afforded cyclic alpha aminophosphonic acids. The convenient chemical conversion of five- and six membered cyclic alpha-amino acids to the corresponding cyclic alpha aminophosphonic acids has been accomplished. PMID- 11383602 TI - New phenolic constituents from the fruit juice of Phyllanthus emblica. AB - Six new phenolic constituents, L-malic acid 2-O- (1), mucic acid 2-O- (5), mucic acid 1,4-lactone 2-O- (6), 5-O- (8), 3-O- (10), and 3,5-di-O- (11) gallates, were isolated from the fruit juice of Phyllanthus emblica together with their methyl esters (2-4, 7, 9), and their structures were determined by spectral and chemical methods. Compounds 5, 6, and 8, the major phenolic constituents of the juice, were present as an equilibrium mixture in aqueous solution. PMID- 11383603 TI - The mechanism of reaction of ebselen with superoxide in aprotic solvents as examined by cyclic voltammetry and ESR. AB - The mechanism of the redox reaction of ebselen with superoxide was investigated using both ESR and electrochemical techniques. The reaction with superoxide in aprotic solvents was followed by means of cyclic voltammetry and ESR spin trapping. A decrease in the oxidation current due to superoxide as a result of the addition of ebselen was clearly observed in the cyclic voltammograms. Ebselen reduced the ESR signal intensity of 5,5-dimethyl-1-pyrroline N-oxide (DMPO) superoxide in a dose-dependent manner. The formation of an amidyl radical in this redox reaction was confirmed by rapid mixing continuous-flow ESR. The selenonate form and the seleninate form of ebselen were identified as the final products of the reaction of ebselen with superoxide. The following mechanism for this redox reaction can be proposed: First, ebselen reacts with superoxide and is converted to an ebselen anion radical; second, the ebselen anion radical reacts with superoxide and is converted to the amidyl radical. Hydrogen abstraction by the amidyl radical occurs and gives both a seleninate form and a selenonate form. PMID- 11383604 TI - Kaempferol acetylrhamnosides from the rhizome of Dryopteris crassirhizoma and their inhibitory effects on three different activities of human immunodeficiency virus-1 reverse transcriptase. AB - Three new kaempferol glycosides, called crassirhizomosides A (1), B (2) and C (3), were isolated from the rhizome of Dryopteris crassirhizoma (Aspidiaceae), together with the known kaempferol glycoside, sutchuenoside A (4). The structures of 1-3 were determined as kaempferol 3-alpha-L-(2,4-di-O-acetyl)rhamnopyranoside 7-alpha-L-rhamnopyranoside, kaempferol 3-alpha-L-(3,4-di-O acetyl)rhamnopyranoside, and kaempferol 3-alpha-L-(2,3-di-O acetyl)rhamnopyranosside-7-alpha-L-rhamnopyranoside, respectively, by chemical and spectroscopic means. Inhibitory effects of 1-4 and kaempferol on human immunodeficiency virus reverse transcriptase-associated DNA polymerase (RNA dependent DNA polymerase and DNA-dependent DNA polymerase) and RNase H activities were investigated. PMID- 11383605 TI - Studies on the constituents of bark of Parameria laevigata Moldenke. AB - One new trimeric proanthocyanidin, epicatechin-(2beta-->O-->7, 4beta-->6) epicatechin-(2beta-->O--->7, 4beta-->8)-epicatechin (5) and two new tetrameric proanthocyanidins, epicatechin-(2beta-->O-->7, 4beta-->8)-[epicatechin-(4beta- >6)]-epicatechin-(4beta-->8)-epicatechin, named as parameritannin A-1 (6), and epicatechin-(2beta-->O-->5, 4beta-->6)-[epicatechin-(2beta-->O-->7, 4beta-->8)] epicatechin-(4beta-->8)-epicatechin, named as parameritannin A-2 (7), have been isolated from the bark of Parameria laevigata Moldenke (Apocynaceae) along with the two known dimers, proanthocyanidin A-2 (1) and proanthocyanidin A-6 (2), and two trimers, cinnamtannin B-1 (3) and aesculitannin B (4). These structures were elucidated by spectral and chemical evidence. PMID- 11383606 TI - A new synthesis of N-alkyl 4-methyl-1,4-dihydropyridines utilizing sec aminodienyl esters with crotonaldehyde. AB - The reactions of sec-aminodienyl esters 3 with crotonaldehyde (4) afforded N alkyl 3-[2-(methoxycarbonyl)ethenyl]-4-methyl-1,4-dihydropyridines 5, providing a new azaelectrocyclization reaction. PMID- 11383607 TI - A novel series of thromboxane A2 synthetase inhibitors with free radical scavenging and anti-peroxidative activities. AB - A novel series of indoline derivatives with imidazole and carboxyl moieties were synthesized and evaluated for their thromboxane A2 (TXA2) synthetase inhibiting, radical scavenging and anti-peroxidative activities. Among the compounds synthesized, 3-[5-substituted-3-[2-(imidazol-1-yl)ethyl]indolin-1-yl]propionic acids showed free radical scavenging activity and inhibitory effects on lipid peroxidation of rat brain homogenate and on arachidonate-induced TXA2-dependent aggregation of rabbit platelets. The anti-platelet and anti-peroxidative activities were related to the lipophilicity of the 5-substituent. The 5-hexyloxy derivative (13) showed about 35-fold higher inhibitory activity on TXA2 synthesis than that of ozagrel and about 100-fold higher activity on lipid peroxidation than that of alpha-tocopherol. Compound 13 showed in vivo anti-thrombotic effect in mice and ex vivo anti-peroxidative activity in rats. PMID- 11383608 TI - Antimalarial and cytotoxic activities of bicycl. AB - Biological evaluations of bicyclo[6.4.0]dodecenone derivatives on antimalarial activity in vitro against Plasmodium falciparum and cytotoxicity against human KB cells were made. (+/-)-(1R*,4S*,7R*,8S*)-4-tert-Butyl-dimethylsiloxy-5,5-dimethyl 1-methyl-9-methylene-7-phenylsulfonylbicyclo[6.4.0]dodec-2,11-dien-10-one (15) exhibited potent antimalarial activity, whereas (+/-)-(1R*,7R*,8S*)-1-methyl-9 methylene-7-phenylsulfonylbicyclo[6.4.0]dodec-2,11-dien-10-one (14) showed significant cytotoxic activity in human KB cells. Both 14 and 15 possess, as a structural character, the exo-methylene moiety in their 6-membered ring of the 8 6 fused ring system. PMID- 11383609 TI - Electron paramagnetic resonance study on free radical scavenging and/or generating activity of dopamine-4-O-sulfate. AB - The free radical scavenging and/or generating activity of dopamine-4-O-sulfate was examined and compared with that of dopamine. In humans, dopamine mostly exists in two isomeric forms of sulfate ester conjugates as metabolites; i.e., dopamine-3-O-sulfate and dopamine-4-O-sulfate in the circulation. Dopamine is generally believed to be oxidized by molecular oxygen or another reactive oxygen species under physiological conditions, to form oxidized dopamine derivatives that are cytotoxic. However, it is not known whether dopamine conjugates are generated on interaction with reactive oxygen species or not. In the present study, we measured the susceptibility to oxidization of dopamine-4-O-sulfate by using electron paramagnetic resonance (EPR) spectroscopy and optical absorption spectrometry. Dopamine was easily oxidized and dopamine-derived radicals appeared, whereas dopamine-4-O-sulfate was not oxidized under physiological conditions. Furthermore, dopamine-4-O-sulfate did not react with a strong oxidizing agent, sodium periodate. These results suggest that dopamine-4-O sulfate has resistance against autoxidation, and seems to be a stable metabolite of dopamine. PMID- 11383610 TI - New cyclopropyl-triterpenoids from the aerial roots of Ficus microcarpa. AB - Four new cyclopropyl-triterpenes, 27-nor-3beta-hydroxy-25-oxocycloartane (1), (22E)-25,26,27-trinor-3beta-hydroxycycloart-22-en-24-al (2), 3beta-acetoxy 15alpha-hydroxy-13,27-cyclours-11-ene (3), 3beta-acetoxy-12alpha-formyloxy-13,27 cycloursan-11alpha-ol (4), together with (23E)-27-nor-3beta-hydroxycycloart-23-en 25-one (5) were isolated from the aerial roots of Ficus microcarpa. Compounds 3 and 4 are rare 13,27-cycloursane-type triterpenes. Their structures were elucidated by spectroscopic and chemical methods. PMID- 11383611 TI - Water-soluble constituents of Glehnia littoralis fruit. AB - From the water-soluble portion of the methanol extract of the fruit of Glehnia littoralis Fr. SCHMIDT ex MIQ. (Umbelliferae; "hamabofu" in Japanese), thirty compounds, including three new monoterpenoids and a new monoterpenoid glucoside, a new benzofuran glucoside, a new alkyl glucoside, and a new glucide, were obtained. Their structures were clarified by spectral investigation. PMID- 11383612 TI - New sterols and triterpenoids from four edible mushrooms. AB - Four edible mushrooms, Panellus serotinus, Lepista nuda, Tricholoma matsutake and Naematoloma sublateritium, have been investigated chemically. Two new sterols, 5alpha,9alpha-epidioxy-(22E)-ergosta-7,22-diene-3beta,6alpha-diol (1) and 5alpha,9alpha-epidioxy-(22E)-ergosta-7,22-diene-3beta,6beta-diol (2), have been isolated from Panellus serotinus. Compound 2 was also isolated from Lepista nuda. A new sterol, 3beta,5alpha,9alpha,14beta-tetrahydroxy-(22E)-ergosta-7,22-dien-6 one (3), and compound 2 have been isolated from Tricholoma matsutake. Three new triterpenoids, sublateriols A-C (4-6), have been isolated from Naematoloma sublateritium. The structures of the new compounds were elucidated on the basis of their spectral data. PMID- 11383613 TI - Effect of mirabilitum in formularization: change of prolyl endopeptidase inhibitory activity and of constituents using the preparation method of Tokaku joki-to. AB - To clarify the effect of Mirabilitum in formularization, change of prolyl endopeptidase inhibitory activity and of constituents using the preparation method of a Kampo formula Tokaku-joki-to ([Japanese characters: see text], Persia and Rhubarb combination) was examined by the liquid chromatography-mass spectroscopy (LC-MS) method. Mirabilitum under boiling condition caused qualitative and quantitative change of the constituents through hydrolysis which caused a change of its activity. This was considered to be the main reason the classical Chinese medical book "Shang han lun ([Japanese characters: see text])" specified that Mirabilitum should be added at a later stage of decoction. PMID- 11383614 TI - Diels-Alder reactions of nitro-2(1H)-pyridones with 2,3-dimethyl-1,3-butadiene. AB - Diels-Alder (DA) reactions of 3- or 5-nitro-2(1H)-pyridones and nitro-2(1H) pyridones containing a methoxycarbonyl group with 2,3-dimethyl-1,3-butadiene were examined. The DA reactions of 3-nitro-2(1H)-pyridones in this paper represent, to the best of our knowledge, the first report of DA reactions of 3-substituted 2(1H)-pyridones and consequent production of isoquinolones. Performing the same reactions with 5-nitro-2(1H)-pyridones yielded quinolones. DA reactions of 2(1H) pyridones with nitro and methoxycarbonyl groups produced isoquinolones, quinolones and phenanthridones (the double DA adducts), aromatized or hydrogenated. The substituent effect was evaluated by calculating the activation energy, using the ab initio MO method. PMID- 11383615 TI - Ethenesulfonamide derivatives, a novel class of orally active endothelin-A receptor antagonists. AB - In the present article we wish to report the discovery of a novel class of ET(A) selective endothelin (ET) receptor antagonists through the modification of the ET(A)/ET(B) non-selective antagonist, Ro47-0203 (Bosentan, 1). Replacement of the benzenesulfonamide group of 1 with a 2-phenylethenesulfonamide group gave compound 5a and resulted in improvement in ET(A)-selectivity. Optimization of the alkoxy side chain attached to the core pyrimidine ring yielded the 2-fluoroethoxy derivative (5n) with further improvement of ET(A)-selectivity. [IC50=2.1 nM for ET(A) receptor, ET(B)/ET(A) ratio=1200]. After oral administration, compound 5n inhibited the big ET-1 induced pressor response in pithed rats with a DR2 value of 2.6 mg/kg and also exhibited a potent antagonistic activity in conscious rats. PMID- 11383616 TI - Co2(CO)8-mediated endo mode cyclization of epoxy-alcohol: synthesis of 2-ethynyl 3-hydroxy-2-methyltetrahydropyran and 2-ethynyl-3-hydroxy-3-methyltetrahydropyran derivatives. AB - Successive treatment of 4,5-epoxy-5-methyl-7-trimethylsilyl-6-heptyne-1-ol with Co2(CO)8 at 0 degrees C and a catalytic amount of BF3 x OEt2 at -78 degrees C gave the tetrahydropyran derivatives with the cobalt-complexed moiety. Similarly 4,5-epoxy-4-methyl-7-trimethylsilyl-6-heptyne-1-ol underwent ring closure under the above conditions to provide the corresponding tetrahydropyran derivatives. The preferential endo mode cyclization over the exo one was observed in these experiments. PMID- 11383618 TI - Resazurin as an electron acceptor in glucose oxidase-catalyzed oxidation of glucose. AB - The behavior of resazurin (1) as an electron acceptor in glucose oxidase (GOD) catalyzed oxidation of glucose under anaerobic conditions is described. When a mixture of 1, glucose, and GOD in phosphate buffer (pH 7.4, 0.1 M) was incubated at 25 degrees C, the resulting solution turned purple to fluorescent pink due to the deoxygenated product, resorufin (2). On incubation of 1 with GOD alone or with H2O2 under essentially the same conditions, no color change was seen, indicating that generation of 2 in the enzymatic reaction is brought about through reduction of 1 by the reduced form (GODred) of GOD, which was also supported by the voltammetric behavior of 1. However, it was found that the enzymatic transformation of 1 to 2 is of no practical use as an indicator reaction for glucose determination using only GOD due to a slow reaction of 1 with GODred. Based on a ping-pong type mechanism with a steady-state approximation, KM and kcat for 1 as an electron acceptor from GODred were estimated to be 15+/-1.3 microM and (5.0+/-0.5) x 10(-2) s(-1), respectively. PMID- 11383617 TI - A prenyloxycoumarin from Psiadia dentata. AB - A new coumarin identified as 5-hydroxy-6-methoxy-7-(3-methyl-but-2-enyloxy)-2H-1 benzopyran-2-one (isoobtusitin) was isolated from Psiadia dentata. This compound showed, in vitro, a moderate inhibitory activity against poliovirus and a very weak activity against (HIV), whereas it was inactive against (HSV1), (VSV), and murine tumoral cell lines (3LL, L1210). PMID- 11383619 TI - Three new triterpene saponins from the seeds of Aesculus chinensis. AB - Three new triterpenoid saponins were isolated from the seeds of Aesculus chinensis, and characterized as 22-tigloylprotoaescigenin 3-O-[beta-D glucopyranosyl (1-->2)] [beta-D-glucopyranosyl (1-->4)]-beta-D glucopyranosiduronic acid (escin IVg, 1), 22-angeloylprotoaescigenin 3-O-[beta-D glucopyranosyl (1-->2)] [beta-D-glucopyranosyl (1-->4)]-beta-D glucopyranosiduronic acid (escin IVh, 2) and 16-angeloyl-21-acetylprotoaescigenin 3-O-[beta-D-glucopyranosyl (1-->2)] [beta-D-glucopyranosyl (1-->4)]-beta-D glucopyranosiduronic acid (escin VIb, 3), together with two known compounds, escin IIIa (4) and desacylescin 1 (5). Their structures were established on the basis of spectroscopic and chemical evidence. PMID- 11383620 TI - Amino acid derivatives with anticonvulsant activity. AB - A series of benzylamides of N-alkylated, N-acylated or free nine cyclic and one linear amino acids as potential anticonvulsants have been synthesized. The structures of the obtained compounds were designed on the basis of the previously determined structure and physicochemical properties/anticvonvulsant activity relationship of the formerly synthesized compounds of this type. The obtained compounds were evaluated in mice after intraperitoneal (i.p.) administration, by maximal electroshock seizure test (MES test), subcutaneous (s.c.) pentylenetetrazol test (s.c. PTZ test) and by the rotarod neurotoxicity test (Tox test). The results were the basis for their classification into one of three classes of the Anticonvulsant Screening Project (ASP) of the Antiepileptic Drug Development Program (ADDP) of the NIH. Three selected compounds were tested quantitatively in rats after oral administration. The MES ED50, s.c. PTZ ED50, Tox TD50 were determined and their protective index (PI) values were calculated. Anticonvulsant activity of the most promising compound (15) was examined in different seizure models. The respective ED50 and PI values of this compound were as follows: against bicuculline, 73 and 1.4; against PTZ, 47 and 2.2; against strychnine, 73 and 1.4; against pilocarpine 156, and 0.7; against kainic acid (2 carboxy-4-isopropenyl-3-pyrrolidineacetic acid), 39 and 2.6; against AMPA (alpha amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid), 10 and 10.3 and against NMDA (N-methyl-D-Aspartic acid), 114 and 0.9. PMID- 11383621 TI - Four new oleanane saponins from Anemone anhuiensis. AB - Four new oleanane triterpene saponins, anhuienosides C-F, together with three known saponins, were isolated from the rhizomes of Anemone anhuiensis (Ranunculaceae). The structures of anhuienosides C-F were elucidated as 3-O-alpha L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl oleanolic acid 28-O-beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3-O-beta-D-xylopyranosyl oleanolic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6) beta-D-glucopyranosyl ester, 3-O-alpha-L-rhamnopyranosyl-(1-->2)-beta-D glucopyranosyl oleanolic acid 28-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranosyl ester, 3-O-beta-D-glucopyranosyl-(1- >3)-alpha-L-rhamnopyranosyl-(1-->2)-beta-D-xylopyranosyl oleanolic acid 28-O alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyranosyl ester, respectively. PMID- 11383622 TI - Studies on the constituents of Clematis species. VIII. Triterpenoid saponins from the aerial part of Clematis tibetana KUNTZ. AB - From the aerial part of Clematis tibetana, two new hederagenin 3,28-O bisdesmosides called clematibetosides A and C, and a new gypsogenin 3,28-O bisdesmoside called clematibetoside B, have been isolated together with ten known saponins. The structures of the new saponins have been elucidated based on chemical and spectral evidence as follows: clematibetoside A, 3-O-(2-O-caffeoyl) beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->4)-beta-D-ribopyranosyl (1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl hederagenin 28-O alpha-L-rhamnopyranosyl-(1-->4)-beta-D-glucopyranosyl-(1-->6)-beta-D glucopyraside; clematibetoside B, 3-O-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L arabinopyranosyl gypsogenin 28-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranoside; clematibetoside C, 3-O-beta-D ribopyranosyl hederagenin 28-O-alpha-L-rhamnopyranosyl-(1-->4)-beta-D glucopyranosyl-(1-->6)-beta-D-glucopyranoside. PMID- 11383623 TI - Synthesis of styrenes through the biocatalytic decarboxylation of trans-cinnamic acids by plant cell cultures. AB - A novel method for producing styrenes from trans-cinnamic acids was developed. When trans-cinnamic acid was incubated with plant cell cultures at room temperature, styrene was obtained. 4-Hydroxy-3-methoxystyrene (2a), 3 nitrostyrene (2f) and furan (2g) were synthesized quantitatively. PMID- 11383624 TI - Radical scavenging activity against 1,1-diphenyl-2-picrylhydrazyl of ascorbic acid 2-glucoside (AA-2G) and 6-acyl-AA-2G. AB - The radical scavenging activity of the stable derivatives, which are O substituted at the C-2 position of ascorbic acid (AA), against 1,1-diphenyl-2 picrylhydrazyl (DPPH) was evaluated in buffer under different pH conditions, and compared with those of AA and alpha-tocopherol. AA was shown to have 50% radical scavenging ability (EC50) at a concentration of 2.2 x 10(-5) M against 0.1 mM DPPH in 60% ethanol. Ascorbyl 6-palmitate, a lipophilic AA derivative which has a free endiol group and is therefore unstable, also showed potent radical scavenging activity with EC50 of 2.9 x 10(-5) M. A typical lipophilic antioxidant, alpha-tocopherol gave a similar EC50 value as that of AA. In contrast, ascorbyl 2,6-dipalmitate, AA 2-phosphate and AA 2-sulfate exhibited negligible scavenging activity. On the other hand, 2-O-alpha-D-glucopyranosyl-L ascorbic acid (AA-2G) and a series of 6-O-acyl-2-O-alpha-D-glucopyranosyl-L ascorbic acids (6-Acyl-AA-2G) themselves exhibited the radical scavenging activity of EC50: 6.1 x 10(-5) M and 4.4 x 10(-5)-5.9 x 10(-5) M, respectively, although their activities were lower than that of AA. Among 6-Acyl-AA-2G derivatives, the EC50 values tended to decrease with increasing length of their acyl carbon group. Increasing pH of the buffer resulted in decrease in the scavenging activity of all compounds tested as expected. We speculate that the difference in the radical scavenging activity of derivatives O-substituted at the C-2 position of AA may be ascribed to the linkage type of the substituent group to the endiol-lactone resonance system and the degree of dissociation of the C-3 proton. PMID- 11383625 TI - Structures of new sesquiterpenoids from Farfarae Flos. AB - Two new bisabolane-type sesquiterpenoids, (3R,4R,6S)-3,4-epoxybisabola-7(14),10 dien-2-one and (1R,3R,4R,5S,6S)-1-acetoxy-8-angeloyloxy-3,4-epoxy-5 hydroxybisabola-7(14),10-dien-2-one, and a new oplopane-type sesquiterpenoid, 14(R)-hydroxy-7beta-isovaleroyloxyoplop-8(10)-en-2-one, were isolated from Farfarae Flos along with three known compounds. The structures of these compounds were elucidated on the basis of spectroscopic evidence. PMID- 11383626 TI - Limonoids from Citrus sudachi. AB - Four new limonoid derivatives, 1-O-methylichangensin (1), sudachinoid A (2), B (3), and C (4) were isolated from the seeds of Citrus sudachi, together with the known compounds ichangensin, obacunone, obacunoic acid and limonin, and the structures of the new compounds were elucidated based on spectroscopic and chemical evidence. PMID- 11383627 TI - New insulinomimetic zinc(II) complexes of alpha-amino acids and their derivatives with Zn(N2O2) coordination mode. AB - Zinc(II) complexes of alpha-amino acids and their derivatives with a Zn(N2O2) coordination mode were found to have in vitro insulinomimetic activity as estimated with the inhibition of free fatty acid release in isolated rat adipocytes treated with epinephrine. It was revealed that the insulinomimetic activities of zinc(II) complexes with over-all stability constants (log beta) less than 10.5 are higher than those of ZnSO4 and VOSO4. The high blood glucose level of KK-Ay mice with type 2 diabetes mellitus was lowered by daily intraperitoneal injections of a zinc(II) complex, cis-[Zn(L-Thr)2(H2O)2], for 14 d. The improvement of diabetes mellitus was confirmed with the oral glucose tolerance test. PMID- 11383628 TI - Conformation of 1,4-dineopentyl-2,5-cis-diphenylpiperazine and its diammonium salts: remarkable change in conformation depending upon the counter anion. AB - The free base of 1,4-dineopentyl-2,5-diphenylpiperazine takes a chair conformation in CDCl3, while the conformation of its diammonium salts changes depending upon the counter anion. PMID- 11383629 TI - Lewis acid-promoted cycloaddition reaction of cyclopropanes with allylsilanes. AB - The treatment of cyclopropanes having donor and acceptor substituents at the vicinal positions on the cyclopropane ring with a Lewis acid readily generates a 1,3-zwitterion, which reacted with allylsilanes to produce cycloadducts and allylic products. It was found that the yield of the cycloadduct depends on the steric demand of the alkyl substituents on the silicon atom. PMID- 11383630 TI - Effects of chlorhexidine digluconate and hydrogen peroxide on Porphyromonas gingivalis hemin binding and coaggregation with oral streptococci. AB - Porphyromonas gingivalis, a gram-negative anaerobe, is one of the major causative agents of periodontal disease. In this study, the effects of chlorhexidine digluconate and hydrogen peroxide on the hemin binding of P. gingivalis and coaggregation of this bacterium with oral streptococci were examined. The pretreatment of P. gingivalis W50 and 381 with chlorhexidine digluconate and hydrogen peroxide increased the hemin binding of these bacteria. The hemin binding of P. gingivalis was increased by the subminimal inhibitory concentration (MIC) of chlorhexidine digluconate. However, concentrations of hydrogen peroxide below the MIC had no effect on the hemin binding of P. gingivalis W50 and 381. Coaggregation of P. gingivalis 381 with Streptococcus oralis ATCC 9811 and Streptococcus gordonii DL1 was diminished by chlorhexidine digluconate. The coaggregation-inhibitory effect was concentration-dependent. Hydrogen peroxide also showed inhibitory effects on the coaggregation of P. gingivalis 381 with S. oralis 9811 and S. gordonii DL1 at concentrations below that used clinically. Concentrations of chlorhexidine digluconate below the MIC inhibited coaggregation. However, concentrations of hydrogen peroxide below the MIC were not effective in reducing the coaggregation of P. gingivalis with oral streptococci. These observations show that chlorhexidine digluconate and hydrogen peroxide could confer variable effects on P. gingivalis hemin binding and coaggregation of this bacterium with oral streptococci. PMID- 11383631 TI - Flexibility and hardness of dental stainless steel wrought wires used in Thailand. AB - Stainless steel wrought wires used as clasp arms for removable partial dentures in Thailand were compared with those used in some other countries (in the as received condition) in terms of flexibility, Vickers microhardness and composition. The results showed that there were significant differences (P< or =0.05) among the wires. A Japanese stainless steel wire (SK) was obviously different from the others. It had the lowest proportional limit and microhardness, but its flexibility was almost the same. The chemical composition of each wire was not greatly different. The wires were about 18-20 wt% chromium and 8-9 wt% nickel, except for the SK wire, which had about 12 wt% nickel. PMID- 11383632 TI - Juvenile xanthogranuloma: case report with immunohistochemical identification of early and late cytomegalovirus antigens. AB - Juvenile xanthogranuloma (JXG) is a dermatological condition of unknown etiology that rarely affects the oral mucosa. There are conflicting reports suggesting that it may represent a reactive virally-induced lesion associated with cytomegalovirus (CMV) infection. The present paper reports an additional case of oral JXG and discusses its possible association with CMV infection. The biotin streptavidin system was used to detect early and late CMV antigens. Positive immunolabelling for both antigens was demonstrated in some histiocytes in the lesion. These findings suggest that JXG may be associated with CMV infection. PMID- 11383633 TI - Titanium casting: the surface reaction layer of castings obtained using ultra-low temperature molds. AB - To examine whether the surface reaction layer of titanium castings can be reduced by lowering the mold temperature during casting, we cast titanium at three mold temperatures, including an ultra-low temperature produced by cooling the mold with liquid nitrogen, then measured the tensile strength and elongation of the castings. The titanium was cast using a centrifugal casting machine, and the molds were incinerated according to the manufacturers' instructions. Castings were then made with the molds at 200 degrees C, 600 degrees C, and an ultra-low temperature (-196 degrees C). The castability of titanium cast in the mold at the ultra-low temperature was good. The Vickers hardness near the surface layer of castings decreased as the mold temperature decreased. PMID- 11383635 TI - Analysis of factors that may affect the enzymatic profile of gingival crevicular fluid: sampling technique, sequential sampling and mode of data presentation. AB - To evaluate the possible effect of sampling technique and sequential sampling on gingival crevicular fluid (GCF) volume and myeloperoxidase (MPO) activity, 14 patients presenting at least two symmetrical maxillary sites with mild/moderate periodontitis were selected. Two sites in each individual were sequentially sampled using either the deep-intracrevicular or orifice technique. Spectrophotometrically determined MPO levels were presented either as total MPO activity or MPO concentration. Although the clinical periodontal status of the 20 sampling sites were similar, the deep-intracrevicular technique regularly provided larger GCF volumes. With both techniques, the last samples contained the highest GCF volume. During sequential orifice sampling, GCF volume was relatively more stable. In general, a depletion of MPO activity was observed with sequential sampling performed with either of the techniques. Depletion of MPO did not replenish to baseline levels at the end of the 10-min sequential sampling. Although MPO activity showed a general reduction during sequential orifice sampling with both modes of data presentation, total MPO activity and MPO concentration did not match with the deep-intracrevicular technique. Due to the potential of affecting GCF volume/composition, the selection of sampling technique seems to be a critical methodological decision in GCF-profile studies, primarily during sequential sampling. In GCF-profile studies, mode of data presentation should also be considered. PMID- 11383634 TI - Influence of environmental conditions on dentin bond strengths of recently developed dentin bonding systems. AB - A study was conducted to investigate the influence of temperature and relative humidity (RH) on the bond strengths of several recently developed dentin bonding systems. Six environmental conditions, (A) 25+/-0.5 degrees C, 50+/-5% RH, (B) 25+/-0.5 degrees C, 80+/-5% RH, (C) 25+/-0.5 degrees C, 95+/-5 % RH, (D) 37+/-0.5 degrees C, 50 +/-5 % RH, (E) 37+/-0.5 degrees C, 80+/-5% RH, (F) 37+/-0.5 degrees C, 95+/-5 % RH were used. Bovine mandibular incisors were mounted in self-curing resin and the facial surfaces were ground on wet #600 SiC paper to expose the dentin. After the tooth surface had been treated according to each manufacturer's instructions, adhesives were applied, followed by condensation of resin composites into a mold placed on the dentin surface. Fifteen specimens per group were stored in distilled water at 37 degrees C for 24 h, and then shear-tested at a cross-head speed of 1.0 mm/min. Statistical analysis was carried out with two way ANOVA followed by Tukey's test (P<0.05). Dentin bond strengths decreased with increasing relative humidity but were not influenced by environmental temperature. Even though one-bottle adhesive systems require a wet dentin surface, their bond strengths are affected by an increase in environmental humidity. PMID- 11383636 TI - Oral health attitudes/behavior and gingival self-care level of Korean dental hygiene students. AB - The purpose of this study was to examine oral health attitudes/behavior and gingival self-care levels of Korean dental hygiene students (n = 271). A questionnaire known as HU-DBI (Korean version) was administered to each class. Higher scores of the HU-DBI indicate better oral health attitudes/behavior. The student's gingival self-care level was scored as excellent (+2), good (+1), questionable (0), poor (-1), or very poor (-2) according to the criteria of the Oral Rating Index (ORI) for youth. The mean score of the HU-DBI was 6.40, and that of the ORI was +0.12. Most samples scored 0 or +1. The mean HU-DBI scores of year-2 and year-3 students (7.06, and 7.61 respectively) were significantly greater than that of year-1 students (5.23) (P<0.001). Twenty-three percent of the students reported a belief that they may eventually require false teeth. Half stated that they put off going to the dentist until they had toothache. Year-1 students were more likely to have this attitude compared to year-3 students. HU DBI scores were significantly associated with ORI scores (r = 0.315; P<0.001). The differences in the HU-DBI and ORI scores across the level of education were highly significant. PMID- 11383637 TI - Pulse irradiation of low-power laser stimulates bone nodule formation. AB - Although low-power laser irradiation provides many anabolic effects such as acceleration of bone formation, the effects of different pulse frequencies used during laser irradiation on bone formation have not been elucidated. Osteoblastic cells isolated from fetal rat calvariae were irradiated once with a low-power Ga Al-As laser (830 nm, 500 mW) in two different irradiation modes; continuous irradiation (CI), and 1 Hz pulsed irradiation (PI). We then investigated the effects on cellular proliferation, bone nodule formation, alkaline phosphatase (ALP) activity, and ALP gene expression. Laser irradiation in both groups significantly stimulated cellular proliferation, bone nodule formation, ALP activity, and ALP gene expression, as compared with the nonirradiation group. Notably, PI markedly stimulated these factors, when compared with the CI group. Since 1 Hz pulsed laser irradiation significantly stimulates bone formation in vitro, it is most likely that pulse frequency is an important factor affecting biological responses in bone formation. PMID- 11383638 TI - Effects of guided bone regeneration with non-resorbable and bioabsorbable barrier membranes on osseointegration around hydroxyapatite-coated and uncoated threaded titanium dental implants placed into a surgically-created dehiscence type defect in rabbit tibia: a pilot study. AB - The purpose of this study was to evaluate the effects of guided bone regeneration (GBR) with either a non-resorbable (ePTFE) or bioabsorbable barrier membrane (RSLT) on osseointegration and extent of bone formation around hydroxyapatite coated (HA) and uncoated threaded titanium (Ti) dental implants placed into surgically-created dehiscence type defects in rabbit tibia. A dehiscence type bone defect, approximately 3 mm in width and height was surgically created on the outer surface of the bone in each tibia of 9 rabbits. For the conventional group, either a HA or a Ti implant was then placed at this site. After the same procedure was performed as in the conventional group, the implant site of the GBR group was covered with either an ePTFE or a RSLT. After 4 months, the rabbits were sacrificed. Specimens were prepared and examined histometrically. It was found that the mean percentage of osseointegration tended to increase in HA compared to Ti implants, both with and without membranes. There was a tendency that the extent of newly regenerated bone was higher in the GBR group than that in the conventional group. PMID- 11383639 TI - Maxillary double lip: report of two cases. AB - Two cases of double lip malformation, an uncommon oral anomaly, are presented, and the factors involved in the development, pathogenesis, diagnosis, and treatment of these uncommon lesions are reviewed. Double lip is usually associated with Ascher's syndrome, which is a rare disease with three more or less consistently associated abnormalities: double upper lip, blepharochalasis and enlargement of the thyroid. Two cases of double lip are reported; one of which was related with Ascher's syndrome, while the other had a traumatic origin. PMID- 11383640 TI - Salivary duct cyst: its frequency in a certain Japanese population group (Tohoku districts), with special reference to adenomatous proliferation of the epithelial lining. AB - It is reported in the European and American literature that salivary duct cysts constitute about 10% of all cysts of the salivary glands, although they appear to be rare in Japan. Between 1975 and 1999, only 3 (0.5%) of 586 salivary gland cysts were diagnosed as salivary duct cysts at the Division of Clinical Pathology, Iwate Medical University Hospital. Histologically, two cases appeared as a unilocular lesion lined by double- and multi-layered epithelium. The other case showed marked, intraluminar and intramural adenomatous proliferation of the epithelial lining, suggesting that the lesion was a benign tumor. A review of the literature yielded only two cases of tumors arising in pre-existing salivary duct cysts. PMID- 11383641 TI - Bronchial atresia: three-dimensional CT bronchography using volume rendering technique. AB - We report a case of asymptomatic bronchial atresia of the anterobasal and laterobasal segments of the right lower lobe. Because of the absence of typical bronchial mucocele on chest radiograph, it was difficult to base the diagnosis on chest radiography alone. Helical CT images demonstrated the characteristic appearance of bronchial atresia, and three-dimensional (3D) CT bronchography using the volume rendering technique helped us to comprehend the spatial relationship between the dilated bronchus distal to the atresia and the more proximal bronchus. PMID- 11383642 TI - CT and MR findings of a pleomorphic adenoma in the peripheral lung. AB - We report a rare case of pleomorphic adenoma in the peripheral lung and its appearance on both CT and MR. The findings correlated well with the pathological findings and corresponded to the nature of the cellular predominance type that is common in the lung but is rare in the salivary gland. PMID- 11383643 TI - A case of adenocarcinoma with mixed subtypes displaying ground-glass opacity on high-resolution CT. AB - We report a case of adenocarcinoma with mixed subtypes with pleural dissemination and lymphatic permeation, although the CT results showed ground-glass opacity that led to the diagnosis of bronchioloalveolar carcinoma without foci of active fibroblastic proliferation. PMID- 11383644 TI - Estimation of time parameter of LQ-model in fractionated radiotherapy of medulloblastoma. AB - PURPOSE: To estimate the dose required to counteract accelerated proliferation, gamma/alpha, of medulloblastoma (MB) cells during fractionated radiotherapy. MATERIALS AND METHODS: Sixty-five patients with MB (local control rate at 5 years: 73%) treated between 1980 and 1990 at our university hospitals were analyzed. The gamma/alpha was estimated by applying the profile likelihood method to Cox's proportional hazards model including the maximum value of time incorporated biologically effective dose, tBEDmax. The tBEDmax was conditionally calculated with various settings of alpha/beta, time to kick off accelerated proliferation, Tk, and gamma/alpha. Pretreatment T-stage, log(age+1), extent of removal, and use of chemotherapy and immunotherapy were also included in the multivariate model. RESULTS: When alpha/beta was taken to be 10 Gy, the estimate of gamma/alpha was 0.52 Gy/day (95% confidence limits: 0.29-0.75) for 0 day of Tk and was 0.55 Gy/day (0.30-0.80) for 21 days of Tk. The gamma/alpha values tended to be larger in the T3 subgroup and in the non-total removal subgroup than in all patients, by about 0.05 Gy/day and 0.1 Gy/day, respectively. When alpha/beta was taken to be 25 Gy, the estimated values were about 0.05 Gy/day smaller than these values. CONCLUSION: The estimated gamma/alpha values were considerably large. Therefore, it is preferable to take this consumed dose per day into consideration in planning and conducting treatments for MB. PMID- 11383645 TI - Basilar artery fenestrations detected by MR angiography. AB - Basilar artery (BA) fenestrations are the most frequently observed fenestrations of the cerebral arteries. Our goal was to examine the magnetic resonance (MR) angiographic incidence, location, and characteristic configuration of BA fenestration. Between April 1996 and March 2000, 600 cranial MR angiographies were performed at our institution. The majority of the patients examined had or were suspected to have cerebrovascular disease. We retrospectively reviewed these 600 MR angiograms. A 1.5 Tesla scanner was used in all studies, and maximum intensity projection (MIP) images obtained using the three-dimensional time-of flight (3D-TOF) technique were displayed stereoscopically. Ten BA fenestrations (1.7%) were detected on MR angiograms. In 87 cases from the early period, the vertebrobasilar (VB) junction was unfortunately not included in the imaging slices. Eight of the 10 fenestrations were located at the proximal BA: six of them were small with a slit-like shape, and two were relatively large with a convex-lens-like shape. A small fenestration was located at the distal BA, and the remaining one was a total duplication of the BA. We stress that knowledge and recognition of BA fenestration are useful and important in the interpretation of cranial MR angiography. Since saccular aneurysms are reported to arise frequently at BA fenestration, the VB junction should be included in the imaging slices of routine cranial MR angiography. PMID- 11383646 TI - Prediction of maximum hepatic enhancement on computed tomography from dose of contrast material and patient weight: proposal of a new formula and evaluation of its accuracy. AB - PURPOSE: To evaluate the accuracy of formulas for predicting maximum hepatic enhancement (MHE) on computed tomography (CT) from patient weight and dose of contrast material. MATERIALS AND METHODS: A total of 104 patients weighing 40-80 kg (mean 58+/-9.7 kg) underwent contrast enhanced abdominal CT. Contrast material [100 ml at 300 mgI/ml (n=54) or 370 mgI/ml (n=50)] was administered at the rate of 2 ml/sec. MHE (HU) was obtained in each patient. We assumed the following exponential model: MHE = C1W(C2)D(C3) (W: body weight in kg, D: dose of iodine in gram, C1, C2, and C3: coefficients). These coefficients were obtained by a multiple regression analysis performed on the data. The accuracy of our formula was evaluated by the ratio (AR) of calculated MHE to measured MHE, and the difference (AD) between calculated MHE and measured MHE. The values of AR and AD in our formula were compared with those in Heiken's formula: MHE=CD/W (C: coefficient). RESULTS: We derived MHE=13.2 D/W(0.5) as our exponential formula and MHE =101 D/W as Heiken's formula. AR was 1.04+/-0.20 (mean +/- SD) and AD was 9.03+/-6.35 with our formula, while the corresponding values were 1.04+/-0.21 and 9.55+/-6.88 with Heiken's formula. The values of AR and AD were not significantly different between the two formulas. CONCLUSION: The combination of our formula and Heiken's formula may be useful in determining the optimal dose of contrast material according to patient weight for the detection of hypovascular liver tumors. PMID- 11383647 TI - Phantom evaluation of scatter and attenuation correction in thallium 201/technetium-99m acquisition in myocardial perfusion single-photon emission computed tomography. AB - PURPOSE: This phantom study was carried out to evaluate the usefulness of scatter correction combined with transmission-based attenuation correction in separate and simultaneous 201Tl/99mTc myocardial SPECT. METHODS: An anthropomorphic torso phantom was used in this study. We used the triple-energy-window (TEW) method for scatter correction and transmission computed tomography (TCT) images for attenuation correction. Images without corrections (UC) and images with corrections (SAC) for scatter and attenuation were reconstructed for the evaluation. RESULTS: The differences in defect size between 99mTc and 201Tl UC images led to interpretation errors in separate (separate protocol) and simultaneous dual-isotope studies (simultaneous protocol). These errors were more prominent in the infero-posterior wall in the simultaneous protocol. Improvement for overestimation in object size and underestimation in defect contrast was visually obtained, and increased contrast was also shown by the myocardium-to defect count (MD) ratios on SAC images in the separate and simultaneous protocols. However, 201Tl SAC images in the simultaneous protocol still had less defect contrast than the corresponding 201Tl SAC images in the separate protocol. CONCLUSIONS: From the results of our phantom experiment, separate rest 201Tl/stress 99mTc-sestamibi acquisitions may be recommended in clinical practice. Further clinical and phantom studies will be needed to validate the method using scatter correction combined with transmission-based attenuation correction. PMID- 11383648 TI - Toxicity outcome after three-dimensional conformal radiotherapy for early stage prostatic cancer. AB - This study evaluated acute and late toxicity in patients with stages of T2a-b, N0, and M0 prostatic carcinoma treated with three-dimensional conformal radiotherapy (3D-CRT). A total of 26 patients were received a mean 69.07 Gy of 3D CRT, and the mean follow-up time was 18.23 months. There were no statistically significant differences between the groups given < or = 70 Gy and > 70 Gy in terms of acute gastrointestinal toxicity, acute genitourinary toxicity, late gastrointestinal toxicity, and late genitourinary toxicity (p>0.05). 3D-CRT offers less normal tissue morbidity with the possibility of giving high doses of irradiation to increase local control. PMID- 11383649 TI - Clinical studies of immunohistochemical staining of DNA-dependent protein kinase in oropharyngeal and hypopharyngeal carcinomas. AB - PURPOSE: DNA-dependent protein kinase (DNA-PK), a serine/threonine kinase composed of p470 catalytic subunit (DNA-PKcs) and p85/p70 heterodimer (Ku antigen), is considered a critical enzyme in the repair of the DNA double-strand breaks (DSB) that are the major lethal lesions induced by ionizing radiation. We investigated the expression of DNA-PK subunits in human tumors. MATERIALS AND METHODS: We examined immunohistochemically the biopsy specimens of 44 patients with oropharyngeal carcinoma and 32 patients with hypopharyngeal carcinoma who had been treated with radiotherapy. RESULTS: Immunopositivity to Ku85 and DNA PKcs was found in all patients. The staining of Ku85 and DNA-PKcs was nuclear, with none of the normal epithelial cells or malignant cells exhibiting cytoplasmic or membrane immunoreactivity. Normal epithelial cells were all stained intensely. In tumors, intense nuclear staining of DNA-PKcs was seen in 75 of 76 tumors, while that of Ku85 was seen in all 76 patients. The radiation responses of a primary tumor that was stained weakly with DNA-PKcs were excellent. CONCLUSION: Our results suggest the possibility of predicting the intrinsic radiosensitivity of human tumors in clinics able to perform immunohistochemical analysis of DNA-PK. PMID- 11383650 TI - Aortoiliac stenooculusive disease and aneurysms: screening with non-contrast enhanced two-dimensional cardiac gated cine phase contrast MR angiography with multiple velocity encoded values and cardiac gated two-dimensional time-of-flight MR angiography. AB - PURPOSE: To evaluate the performance of two-dimensional cine phase contrast MRA with multi-velocity encoded values (multi-VENC cine PC) and ECG-gated two dimensional time-of-flight MRA (ECG-2D-TOF) for the detection of stenoocclusive lesions and aneurysms in the aortoiliac area, when each method was used individually and when the two methods were used together. METHODS: Forty-one patients were included in this study. Multi-VENC cine PC and ECG-2D-TOF were obtained first, then contrast enhanced three-dimensional magnetic resonance angiography (CE-3D-MRA) was performed as the standard of reference. Two observers reviewed the images separately without knowledge of patients' symptoms or histories. Sensitivities and specificities were obtained separately for stenooclusive lesions and aneurysms by two reviewers. RESULTS: When the two methods were applied together, high sensitivities (93.0 by observer 1 and 91.9% by observer 2) and adequate specificities (87.6 and 82.3%) were obtained for stenoocclusive lesions. For aneurysms, moderate to high sensitivities (91.1 and 71.1%) and high specificities (98.8 and 99.4%) were obtained. CONCLUSION: These results suggest that the performance of two non-contrast enhanced MRA techniques may be valuable as a screening tool when the two methods are applied together. PMID- 11383651 TI - Plasmid and chromosomal basis of tolerance to cadmium and resistance to antibiotics in normal bovine duodenal bacterial flora. AB - Cadmium (Cd) tolerance and antibiotic resistance was studied in duodenal flora of 20 normal bovine samples. Twelve bacterial isolates (5 Staphylococcus spp, 4 Enterococcus faecalis, 2 Bacillus spp, and a Pseudomonas sp) were grown in Luria broth containing 0.05 to 0.8 mM of cadmium chloride (CdCl). All isolates displayed multiple antibiotic resistance, with 2 Enterococcus strains and Pseudomonas pickettii demonstrating resistance to 12/17 antibiotics tested. With the exception of Staphylococcus sp, all contained plasmid DNA. Curing to remove plasmid DNA determined if Cd tolerance and/or antibiotic resistance was plasmid or chromosomally mediated. None of the bacteria became sensitive to CdCl after curing, suggesting that tolerance was not plasmid-mediated. Six bacteria became sensitive to antibiotics after curing indicating that antibiotic2 resistance was plasmid mediated. Two of these bacteria became sensitive to multiple antibiotics; a Staphylococcus sp became sensitive to ampicillin, ceftiofur and cephalothin, and a Enterococcus strain became sensitive to neomycin, oxacillin, and tiamulin. All of the isolates were probed for the presence of known Cd-resistance genes (cadA, cadC, and cadD). DNA-DNA hybridization revealed cadA- and cadC-related sequences in chromosomal DNA of a Staphylococcus sp, an Enterococcus strain, and in plasmid DNA of another Staphylococcus sp. No cadD-related sequences were detected in any of the 12 isolates even under reduced stringency of hybridization. PMID- 11383652 TI - Chronic toxicity of dietary 2,3,7,8-tetrachlorodibenzo-p-dioxin to mink. AB - Mature female natural dark mink (Mustela vison) were fed 0.0006 (control), 0.016, 0.053, 0.180, or 1.40 ppb 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) for 131-132 d to ascertain the chronic toxic effects of TCDD in mink, including reproduction. Consumption of the 1.4 ppb TCDD diet resulted in lethargy, bloody stools, and 16.7% mortality. Final mink body weights were inversely proportional to the dietary TCDD concentrations. Due to subnormal mink breeding, definitive effects of TCDD on mink reproductive performance were not ascertained; however, there were significant dose-dependent decreases in kit (young mink) birth weight and survival from birth to 3 w of age in the groups that had reproduction. There were also significant differences in adult minkwhite blood cell counts, plasma total solids, serum iron, phosphorus, albumin, total protein, total CO2, cholesterol, osmolality, and anion gap concentrations, and alanine aminotransaminase activity between the various dietary groups. During the latter stages alopecia and thickened, deformed, and elongated toenails were observed in the adult mink fed 1.4 ppb TCDD. At termination the mink fed 1.4 ppb TCDD had ascites, gastric ulcers, intestinal hemorrhages, depletion of adipose tissue, and mottled and/or discolored livers, spleens, and kidneys. Focal lymphocytic meningitis in region of the olfactory bulb was present in 42% of the mink fed 1.4 ppb TCDD. These results confirmed the high sensitivity of mink to TCDD and revealed a toenail abnormality not previously reported for mink fed TCDD. PMID- 11383653 TI - Correlation of endophyte toxins (ergovaline and lolitrem B) with clinical disease: fescue foot and perennial ryegrass staggers. AB - Fescue foot, summer syndrome, reproductive problems, and ryegrass staggers are all diseases of livestock related to endophyte toxins in pasture grasses. Range finding experiments and case studies of fescue foot relative to ergovaline toxin found in endophyte infected tall fescue and lolitrem B present in endophyte infected perennial ryegrass were conducted. Within 42 d of initiating a feeding trial with chopped tall fescue straw containing 825 ppb ergovaline and at environmental temperatures of 15.9 C clinical signs of fescue foot were seen in cattle. Sheep on tall fescue pastures in November consuming feed with 540 ppb ergovaline and at environmental temperatures of 7.8 C developed fescue foot in 21 d while sheep on the adjacent field in the previous 2 mo with environmental temperatures of 16.6 C and 12.8 C and 458 ppb ergovaline in the pasture grasses did not. In a field outbreak of fescue foot affecting 42/425 feeder lambs in November, the ergovaline of sample pasture grasses had a mean concentration of 813 ppb. Perennial ryegrass staggers was seen in 42/237 feeder lambs when mean lolitrem B in the sampled grass was 2,135 ppb. Overgrazing both tall fescue and ryegrass fields increased probability of clinical disease since the highest levels of toxin were found in the crowns and basal leaf sheaths of tall fescue and perennial ryegrass respectively. Based on these findings, ergovaline dietary levels of 400 to 750 ppb to cattle and 500 to 800 ppb to sheep and lolitrem B levels of 1,800 to 2,000 ppb in feed for both species are approximated threshold values for disease. Cold environmental temperatures are equally important to toxin concentrations in precipitating fescue foot disease. PMID- 11383654 TI - Interplant alkaloid variation and Senecio vernalis toxicity in cattle. AB - Senecio vernalis and other plants containing pyrrolizidine alkaloids (PA) are implicated in the poisoning of cattle. The liver is a known target organ. In this study the content of the alkaloids senecionine (SCO), senkirkin (SKK) and seneciphyllin (SCP) and their toxic effects in cattle were studied. The content of these 3 compounds only varied by a factor of 2 within 10 plant collections at different locations in western Denmark (Jutland). However, individual alkaloids varied 3-fold, and the interplant variation for some of the PA up to 8-fold. SCO and SKK had very short half lives, 20 min and 70 min respectively. In cattle fed dried plant material corresponding to 200 and 400 g of fresh material for 10 d alanine aminotransferase, alkaline phosphatase and g-glutamyl transferase activities remained unchanged. Cattle subsequently fed fresh plant material up to 1 kg/d for 8 d also had no change in liver enzyme activities. Cattle did not show any clinical signs of poisoning, and no morphological liver changes were observed. PMID- 11383655 TI - Toxicity of dietary Heliotropium dolosum seed to mice. AB - Experimental pyrrolizidine alkaloid intoxication was produced in inbred Swiss mice. Animals were fed diets containing 0, 1, 3, 5, or 10% Heliotropium dolosum seed for 24 w. The seeds contained 0.13% total alkaloid concentration composed of 4 specific components: lasiocarpine (78.79%), heliosupine (11.96%), echimidine (5.43%), and heliotrine (3.82%). Deaths occurred in all dosed groups and increased with dietary seed concentration. Massive to submassive liver necrosis together with sinusoidal congestion, and hemorrhage or multifocal hepatocytic necrosis was limited to animals which survived < 5 w and were fed on 10% seed. Moderate to severe hepatomegalocytosis, scattered single cell necrosis, and mild subcapsuler fibrosis were seen in all test group animals that survived > 5 w. Intranuclear eosinophilic inclusions in hepatocytes and bile duct and ductular cell hyperplasia were the most noticeable lesions in the 1, 3, and 5% groups. There was mild to moderate renal tubular megalocytosis in the 3, 5, and 10% groups. It seems likely that H dolosum seed, at least to a limited extent, constitutes a health hazard for certain animal species. PMID- 11383656 TI - Pteridium aquilinum (bracken fern) toxicity in cattle in the humid Chaco of Tarija, Bolivia. AB - We studied the toxicity caused by chronic ingestion of Pteridium aquilinum (bracken) in cattle in the humid Chaco of Tarija, Bolivia. Bovine enzootic haematuria (BEH) and Carcinoma of the esophagus (CE) affected the herds. Sick animals showed caquexia, anemia, leucopenia and urine that turned from pink to intense red color with the presence of blood clots. Cattle grazed in the humid forests of the mountains where P aquilinum represented more than 50% of the plants. P aquilinum var arachnoideum and P aquilinum var Caudatum were present. Toxic norsesquiterpene, ptaquiloside, was identified in both varieties. Carcinomas were in the urinary bladders and esophagus of 100% and 50% of the cattle, respectively. Control of the intoxication could be difficult due to geographic characteristics of this Bolivian region. PMID- 11383657 TI - Metabolic effects of metaproterenol overdose: hypokalemia, hyperglycemia and hyperlactatemia. AB - This case report of metaproterenol toxicity was associated with hypokalemia, hyperglycemia and hyperlactatemia. A similar triad has been reported in acute theophylline poisoning. Hypokalemia and hyperglycemia have been reported with other beta-agonists, but not metaproterenol. Patients presenting with the metabolic triad of hypokalemia, hyperglycemia, and hyperlactatemia should have theophylline and beta-agonist toxicity included in their differential diagnosis because the prognosis and therapy of these 2 intoxications is markedly different. PMID- 11383658 TI - Acute renal failure following latrodectism. AB - Latrodectism is considered dangerous for human beings. Acute renal failure after envenomation is not common and usually results from prerenal failure. We report a 59-y-o man with acute oliguric renal failure due to a combination of prerenal and renal causes after being bitten by a black Latrodectus spider. He had the characteristic anxiety, severe hypertension, tremor, facial edema, and generalized diaphoresis. The patient recovered within a week without sequelae. Clinicians should not overlook the possibility of acute renal failure in latrodectism. PMID- 11383659 TI - High-dose loratadine exposure in a six-year-old child. AB - Loratadine is a long-acting antihistamine indicated for the treatment of seasonal allergic rhinitis and chronic idiopathic urticaria in patients 6 years of age and older. The literature contains little information on high-dose loratadine exposures; as a consequence, poison centers are unsure of the loratadine dose that can be managed with observation and the dose that requires treatment. We report an intentional ingestion of 300 mg loratadine by a 6-y-old child that resulted in minor elevation of blood pressure and heart rate when managed with supportive care only. Further studies and case series are needed before a minimum toxic dose can be established. PMID- 11383660 TI - Atypical rumen acidosis in a dairy herd from whiskey distillery by-products. AB - Decreased milk and reproductive performance, high incidence of gastrointestinal surgeries, and acute deaths were investigated in a herd of Holstein cows. The health problems were due to abnormally low rumen pH's from ingestion of 30 gal/hd/d of a 3.4 pH liquid feed ingredient. A combination of acid neutralizing agents (calcium hydroxide plus sodium carbonate) alleviated the toxic effects of the feed ingredient. PMID- 11383661 TI - Intoxication of goats by Plumbago scandens in Northeastern Brazil. AB - In the State of Paraiba, Northeastern Brazil, goat mortality occurred with the ingestion of Plumbago scandens. The fresh plant was then given experimentally to 4 goats at 5, 10, 17.5 or 25 g/kg bw. Depression, anorexia, salivation with foamy saliva, bellowing, bruxism, humpbacked posture, bloat, ruminal atony, continuous lateral head movements, tachycardia, dyspnea and dark brown to black urine were observed in the goats given 17.5 or 25 g/kg bw. The goats receiving 5 or 10 g/kg bw had less severe signs. The goat dosed with 25 g/kg bw died after 18-20 h. All others recovered in 3-9 d. At necropsy of the high dose goat, the main lesions were dark violet to black discoloration of the mucosa of the tongue, esophagus, reticulum and ventral sac of the rumen, and gelatinous edema in the visceral ruminal peritoneum. Histologically the reticulum and ventral rumen sac had diffuse epithelial necrosis and severe edema and neutrophilic infiltration of the submucosa. Separation of the ruminal epithelium from the submucosa was observed. Epithelial degeneration and necrosis was also seen in the omasum, esophagus and tongue. Reproduction of the disease with clinical signs similar to those observed by the farmer in the spontaneously affected goats suggests that the clinical mortality was caused by ingestion of Plumbago scandens. PMID- 11383663 TI - The potential toxicity of Ilex myrtifolia in beef cattle. AB - Spatial and temporal patterns of death loss in a beef cowherd indicated a possible relationship between the loss of 11 cows and the consumption of Ilex myrtifolia (myrtle leaf holly). To investigate this relationship, plant material from Ilex myrtifolia was harvested and 2 feeding trials were performed. The 1st trial involved intermittent feeding of plant material to 4-mo-o calves for 2 w, and the 2nd trial was continuous plant feeding to 2-mo-o calves for 35 d. No significant clinical pathology, histological or gross lesions resulted and no clinical signs consistent with the original herd problem were observed, suggesting that berries, leaves and stems from Ilex myrtifolia were not sufficiently toxic to induce clinical effects under these experimental conditions. PMID- 11383662 TI - Dietary exposure to 3,3',4,4',5-pentachlorobiphenyl (PCB 126) or 2,3,7,8 tetrachlorodibenzo-p-dixon (TCDD) does not induce proliferation of squamous epithelium or osteolysis in the jaws of weanling rats. AB - Previous studies demonstrated that dietary exposure to 24 ppb 3,3',4,4',5 pentachlorobiphenyl (PCB 126) or 2.4 ppb 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced maxillary and mandibular proliferation of periodontal squamous epithelium, osteolysis of alveolar bone, and loose and displaced teeth in juvenile mink (Mustela vison). This study determined if such effects could be induced in laboratory rats. Feeding weanling male Long Evans rats 20 or 100 ppb PCB 126 or 1 or 10 ppb TCDD for up to 101 days caused a dose-dependent decrease in body weight gains but did not produce the jaw lesion observed in PCB 126- or TCDD-treated mink. PMID- 11383664 TI - Diazepam poisoning with one-month monitoring of diazepam and nordiazepam blood levels. AB - A 54-y-old man ingested 2 g of bulk laboratory diazepam and was treated with activated charcoal, enhanced diuresis and flumazenil infusion. The treatment resulted in awakening, but the patient had drowsiness, dysarthria, diplopia, and dizziness for 9 d. Blood levels of diazepam and its main metabolite, nordiazepam, were obtained for 1 mo. The half-lives in this benzodiazepine overdose were longer than those seen with therapeutic doses. Benzodiazepines should not be readministrated when patients awake after suicide attempts. PMID- 11383665 TI - Tissue cadmium and locomotor behavior following acute inhalation exposure to cadmium aerosol in rats. AB - Rats were exposed for 1 h to increasing concentrations of cadmium (Cd) through inhalation of cadmium chloride aerosol using nose-only inhalation chambers and depositions of Cd in lungs, liver and kidneys were measured. Changes in spontaneous locomotor activities were recorded 2 and 7 d after cessation of exposure. A concentration dependent increase in Cd in lungs, liver and kidneys was accompanied by significant alterations in spontaneous locomotor response that was dependent on the air Cd concentrations and the postexposure duration. The study shows decreased spontaneous locomotor activity due to Cd accumulation in tissues. PMID- 11383666 TI - An unusual presentation of opioid-like syndrome in pediatric valproic acid poisoning. AB - We report a 3-y-o boy who accidentally poisoned himself with valproic acid (VPA). Clinical features included profound coma, depressed respiration and miosis. Treatment included naloxone, gastric lavage, and activated charcoal and a saline cathartic. The patient fully recovered and was discharged 24 h after the admission. Prompt use of naloxone is advised whenever the triad of coma, pinpoint pupils and depressed respiration concur with the clinical possibility of VPA intoxication. PMID- 11383667 TI - Spin doctors of the world--take heed! PMID- 11383668 TI - Management of anticoagulant poisoning--author's response. PMID- 11383669 TI - Technology integration for the provision of population-based equitable patient care: The Tayside Regional Diabetes Network--a brief description. PMID- 11383670 TI - Measuring differences in the provision of diabetes care. PMID- 11383671 TI - Establishing the UK National Paediatric Diabetes Audit. PMID- 11383672 TI - PROSIT--a quality management system for diabetic patients with nephropathy in Germany. PMID- 11383673 TI - Confidentiality and diabetes registers. PMID- 11383674 TI - Hyperglycemia and QT interval: time for re-evaluation. AB - Among the many mechanisms proposed to explain the relationship between glucose levels and subsequent cardiovascular events, a prolonged QT interval, ie the time interval between the start of activation of the ventricle and completion of its repolarization, seems noteworthy. In Type 2 diabetic patients, for example, the prevalence of QTc (corrected QT interval) prolongation is as high as 26% and is associated with heart disease. The mechanism by which hyperglycemia may produce ventricular instability, as manifested in QTc prolongation, may be increased sympathetic activity, increased cytosolic calcium content in myocytes, or both. By raising the production of free radicals, high glucose may reduce nitric oxide (NO) availability to target cells inducing a state of increased vasomotor tone and ventricular instability. Reduction of Na+/K+-ATPase activity, inhibition of Ca2+-ATPase activity, depressed Na+/Ca2+ exchanger activity, and activation of Na+/H+ antiport may all be implicated. Further studies are urgently needed to characterize in full the effect of hyperglycemia on vascular cells, in order to find therapeutic approaches that lessen the burden of cardiovascular morbidity and mortality in human diabetes. PMID- 11383675 TI - Monitoring renal function in obese patients with type 2 diabetes mellitus in daily practice. AB - BACKGROUND: Endogenous creatinine clearance (GFR(24-hr)) and the Cockcroft formula (GFR(Cockcroft)) are used for the assessment of the glomerular filtration rate (GFR) in daily practice. The influence of extreme obesity in Type 2 diabetes mellitus (T2DM) on the Cockcroft formula was evaluated. METHODS: We compared GFR as calculated by GFR(Cockcroft) to GFR as determined by GFR(24-hr) in 210 patients with T2DM and normal serum creatinine levels. The influence of body mass index (BMI) on the difference between both methods was evaluated. RESULTS: GFR(Cockcroft) was an overall good predictor for GFR(24-hr), but a large individual difference between both methods was observed. A significant correlation was found between the two methods (r=0.962, p<0.001). In T2DM patients with a BMI>35 kg/m2, mean GFR(Cockcroft) was 18% greater than GFR(24 hr). For every unit BMI in this patient group the GFR(Cockcroft) increased by 7.9% relative to the GFR(24-hr). CONCLUSIONS: The major limitation of this evaluation is the lack of a gold standard for GFR measurement. This evaluation of daily practice shows the GFR estimation by the Cockcroft formula and the 24-hr creatinine clearance are in agreement, although a large individual variation was found between both methods making the interchangeability questionable. In view of the influence of BMI on the Cockcroft formula, this should not be used to estimate the GFR in T2DM patients with extreme obesity. PMID- 11383676 TI - Vitamin E intake reduces plasminogen activator inhibitor type 1 in T2DM patients. AB - Previous studies hypothesised that vitamin E could protect against coronary heart disease and vascular complications in diabetes, but no studies have been performed regarding its eventual effects on fibrinolysis. Nevertheless, in Type 2 diabetes mellitus (T2DM) a profound reduction in the fibrinolytic activity has been demonstrated to be involved in vascular complications, probably due to plasminogen activator inhibitor type 1 (PAI-1) overproduction. On this basis we aimed to verify whether an antioxidant treatment with vitamin E is able to lower PAI-1 plasma levels in T2DM. Thirteen T2DM patients (9 males and 4 females; mean age+/-SD, 64.4+/-3.3 yr) were selected through strict admission criteria. These patients were treated with vitamin E (500 IU/die) for 10 weeks. Glyco lipometabolic, oxidative and haemocoagulative parameters were evaluated at baseline and after 5, 10, 30 and 60 weeks. Vitamin E levels at different times were [median (interquartile range)] 6.1 (5.3-7.7), 8.5 (7.3-9.9), 9.7 (8.9-12.9), 5.6 (4.4-6.8), 5.7 (4.5-7.1) microg/ml, respectively. Significant differences were found for PAI-1 antigen (p=0.006), PAI-1 activity (p=0.028), apolipoprotein B (p=0.015) and antioxidant defence, evaluated as ferric reducing ability of plasma (FRAP) values (p=0.005). Particularly, decrements were detected for PAI-1 antigen between baseline and the 10th week (p<0.05), followed by an increase back to basal at the 30th week. Similar behaviour was found for PAI-1 activity. Regarding the antioxidant defence, FRAP values increased until the 30th week (p<0.05) with a decrease at the 60th week. These results demonstrate that vitamin E is able to lower PAI-1 levels in diabetic patients but this effect does not seem related to improvements of glycometabolic data or to the increase in FRAP values, suggesting that PAI-1 overproduction can be decreased by other effects of vitamin E on endothelial cells. PMID- 11383677 TI - Vitamin D deficiency reduces insulin secretion and turnover in rats. AB - Vitamin D deficiency can result in low serum insulin levels, which lead to impaired glucose tolerance. The present study was conducted to assess the mechanisms of this impairment. Pancreatic secretion of insulin and insulin turnover in vitamin D-deficient rats were studied. Insulin secretion was found to be significantly decreased in vitamin D-deficient rats compared to control rats. Supplementation with calcium alone could reverse the defect. The impaired insulin secretion observed in vitamin D deficiency was not specific to glucose stimulation but was also seen in response to arginine. Insulin turnover was studied in vivo using A14-(125)I-insulin. The pharmacokinetic parameters (elimination rate constant K, insulin biological half-life, t1/2, plasma concentrations of insulin at time zero I0) were calculated from the plasma radioactivity. The vitamin D-deficient rats had a lower degradation rate than controls. These findings suggest that vitamin D deficiency leads to reduction in both insulin secretion and turnover, and that the reduced turnover could be an adaptive mechanism to conserve the low amounts of insulin secreted. PMID- 11383678 TI - A review of European experience with aggregated diabetes databases in the delivery of quality care to establish a future vision of their structure and role. PMID- 11383679 TI - Delivery of high-quality care depends on an integrated electronic medical record. PMID- 11383680 TI - A standard architecture for sustainability and integration of care. PMID- 11383681 TI - Quality development in diabetes care. PMID- 11383682 TI - PROMODR: Progressive Model for Diabetes Register. PMID- 11383683 TI - Atmospheric pressure chemical ionization mass spectrometry for analysis of lipids. AB - Atmospheric pressure chemical ionization (APCI) mass spectrometry (MS) has proven to be a very valuable technique for analysis of lipids from a variety of classes. This instrumental method readily produces useful ions with gentle fragmentation from large neutral molecules such as triacylglycerols and carotenoids, which are often difficult to analyze using other techniques. Molecules that are easily ionized, such as phospholipids, produce molecular ions and diagnostically useful fragment ions that are complementary to those produced by methods such as electrospray ionization MS with collision-induced dissociation. The simplicity and versatility of APCI-MS make it an ideal tool for use in solving hitherto very difficult analytical problems. PMID- 11383684 TI - Tolerance and incorporation of a high-dose eicosapentaenoic acid diester emulsion by patients with pancreatic cancer cachexia. AB - Chemotherapy and radiotherapy offer little benefit to patients with advanced pancreatic cancer. Eicosapentaenoic acid (EPA) has anticancer effects both in vitro and in animal models. The dose of EPA that can be administered to cancer patients has previously been limited by the low purity of available preparations and the tolerability of large capsules. A high-purity preparation of EPA as a 20% oil-in-water diester emulsion allowed a small study of the tolerance, incorporation, and effects of EPA in high doses in five patients with advanced pancreatic cancer. Patients underwent assessment at baseline and every 4 wk thereafter. All patients managed to tolerate a dose providing 18 g EPA per day, with doses between 9 and 27 g daily being taken for at least a month. Dosage was limited by a sensation of fullness, cramping abdominal pain, steatorrhea, and nausea. All such symptoms were controlled by dose reduction or pancreatic enzyme supplements. No other adverse effects attributable to the trial agent were observed. Plasma phospholipid EPA content increased from around 1% at baseline to 10% at 4 wk and 20% at 8 wk. Incorporation of EPA into red blood cell phospholipids reached levels of around 10%. The present study has shown that a novel, high-purity, EPA diester emulsion can be tolerated at a dose providing around 18 g EPA per day with side-effects being easily controlled. The acceptibility of large doses of oral EPA should allow larger controlled clinical studies into potential anticancer effects of EPA. PMID- 11383685 TI - Effects of carp and tuna oils on 5-fluorouracil-induced antitumor activity and side effects in sarcoma 180-bearing mice. AB - In this study, we examined the effects of fish oils on 5-fluorouracil (5-FU) induced antitumor activity in mice. First, we examined the antitumor activity of the oral administration of two fish oils (carp oil and tuna oil) in sarcoma 180 bearing mice. Carp oil (0.2 and 0.4 mL/mouse) and tuna oil (0.2 and 0.4 mL/mouse) had no effects on tumor growth. Next, we examined the combined effects of 5-FU plus two fish oils (carp oil and tuna oil) on the antitumor activity and side effects compared to the effects of 5-FU alone (12.5 mg/kg/d). We found that carp oil (0.4 mL/mouse) or tuna oil (0.2 or 0.4 mL/mouse) enhanced the ability of 5-FU (12.5 mg/kg/d) to prevent tumor growth, without increasing side effects such as myelotoxicity and immunocompetent organ toxicity. Tuna oil (0.2 mL/mouse) slightly reduced body weight as compared to the effects of 5-FU alone and water alone (control). The area under the curve (AUC) (0-120 min) of blood 5-FU levels was reduced by the oral co-administration of 5-FU with carp oil or tuna oil. Apparent Tmax was shortened by the oral co-administration of 5-FU with carp oil or tuna oil. On the other hand, AUC (0-4 h) of 5-FU incorporation into tumor RNA fraction was not affected by the oral co-administration of 5-FU with carp oil or tuna oil. PMID- 11383686 TI - Oxidative stability of low density lipoproteins and vitamin E levels increase in maternal blood during normal pregnancy. AB - In 24 healthy pregnant women, parameters related to the oxidative stability of low density lipoproteins (LDL) were determined at three times during pregnancy and shortly after delivery. The fatty acid composition of plasma phospholipids (PL) and the plasma concentrations of vitamin E, vitamin A, and beta-carotene were assessed in the same samples. Total triglyceride (TG), total cholesterol, LDL-cholesterol, and high density lipoprotein (HDL)-cholesterol concentrations were also determined. The length of the lag phase of isolated LDL challenged with Cu2+ ions significantly increased with the progression of pregnancy. The oxidation rate and the amount of conjugated dienes formed increased and reached a maximum at 29-37 wk of pregnancy. Total TG, cholesterol, and LDL-cholesterol reached a maximum in the third trimester of pregnancy. beta-Carotene remained stable, vitamin A decreased, and vitamin E significantly increased throughout pregnancy. Vitamin E plasma concentration correlated positively with the length of the lag phase. The increased levels of vitamin E could contribute to the higher resistance of LDL toward oxidation with progressing gestation, measured by the prolonged lag phase. Furthermore, vitamin E plasma levels correlated positively with TG concentration but not with LDL-cholesterol. The level of polyunsaturated fatty acids in PL decreased with the progression of pregnancy. No correlation was found between the fatty acid composition of plasma PL, nor with the cholesterol concentration, and the parameters studied related to the oxidative stability of LDL. The major finding of this study is the increased oxidative resistance of LDL with progressing gestation. PMID- 11383687 TI - Oxidative catabolism of alpha-tocopherol in rat liver microsomes. AB - The goal of this study was to clarify the mechanism responsible for the catabolism of alpha-tocopherol. The vitamin, bound to albumin, was incubated with rat liver microsomes and appeared to be broken down. Optimal production of the metabolite was obtained when 1 mg of microsomal protein was incubated with 36 microM of alpha-tocopherol in the presence of 1.5 mM of NADPH. Chromatographic and mass spectrometric analyses of the metabolite led to the conclusion that it consists of an omega-acid with an opened chroman ring, although we could not perform nuclear magnetic resonance analysis to confirm this. Our data show that alpha-tocopherol is omega-oxidized to a carboxylic acid and that this process can occur in rat liver microsomes in the presence of NADPH and O2. The oxidation to the quinone structure appears to be a subsequent event that may be artifactual and/or catalyzed by a microsomal enzyme(s). PMID- 11383688 TI - Lipid and fatty acid profiles in rats consuming different high-fat ketogenic diets. AB - High-fat ketogenic diets are used to treat intractable seizures in children, but little is known of the mechanism by which these diets work or whether fats rich in n-3 polyunsaturates might be beneficial. Tissue lipid and fatty acid profiles were determined in rats consuming very high fat (80 weight%), low-carbohydrate ketogenic diets containing either medium-chain triglyceride, flaxseed oil, butter, or an equal combination of these three fat sources. Ketogenic diets containing butter markedly raised liver triglyceride but had no effect on plasma cholesterol. Unlike the other fats, flaxseed oil in the ketogenic diet did not raise brain cholesterol. Brain total and free fatty acid profiles remained similar in all groups, but there was an increase in the proportion of arachidonate in brain total lipids in the medium-chain triglyceride group, while the two groups consuming flaxseed oil had significantly lower arachidonate in brain, liver, and plasma. The very high dietary intake of alpha-linolenate in the flaxseed group did not change docosahexaenoate levels in the brain. Our previous report based on these diets showed that although ketosis is higher in rats consuming a ketogenic diet based on medium-chain triglyceride oil, seizure resistance in the pentylenetetrazol model is not clearly related to the degree of ketosis achieved. In combination with our present data from the same seizure study, it appears that ketogenic diets with widely differing effects on tissue lipids and fatty acid profiles can confer a similar amount of seizure protection. PMID- 11383689 TI - Energy value and digestibility of dietary oil containing mainly 1,3 diacylglycerol are similar to those of triacylglycerol. AB - Diacylglycerol (DAG) is a component of various vegetable oils. Approximately 70% of the DAG in edible oils are in the configuration of 1,3-DAG. We recently showed that long-term ingestion of dietary oil containing mainly 1,3-DAG reduces body fat accumulation in humans as compared to triacylglycerol (TAG) oil with a similar fatty acid composition. As the first step to elucidate the mechanism for this result, we examined the difference in the bioavailabilities of both oils by measuring food energy values and digestibilities in rats. Energy values of the DAG oil and the TAG oil, measured by bomb calorimeter, were 38.9 and 39.6 kJ/g, respectively. Apparent digestibility expressed according to the formula: (absorbed) x (ingested)(-1) x 100 = (ingested - excreted in feces) x (ingested)( 1) x 100 for the DAG oil and the TAG oil were 96.3+/-0.4 and 96.3+/-0.3% (mean +/ SEM), respectively. The similarity in the bioavailabilities of both oils supports the hypothesis that the reduced fat accumulation by dietary DAG is caused by the different metabolic fates after the absorption into the gastrointestinal epithelial cells. PMID- 11383690 TI - Nonalcoholic components in wine reduce low density lipoprotein cholesterol in normocholesterolemic rats. AB - Using an experimental model that enables the effects of alcohol to be distinguished from the effects of the nonalcoholic components present in wine, we determined whether wine has effects other than those of alcohol on the metabolism of cholesterol. Male rats were fed a standard diet and had free access to water and either wine or an equivalent alcohol solution for 45 d or 6 mon. Alcohol intake was similar in the two groups of animals. Consumption of the alcohol solution or wine did not influence plasma cholesterol or high density lipoprotein cholesterol. At 45 d, the consumption both of wine and of alcohol solution reduced low density lipoprotein (LDL)-cholesterol and very low density lipoprotein cholesterol. At 6 mon, only the rats that consumed wine had reduced LDL-cholesterol. After 45 d of consuming alcohol solution, total cholesterol in the aorta was significantly increased mainly as a result of the rise in free cholesterol. In the aorta, the effect of wine consumption was similar to the effect of alcohol solution consumption, although it was less intense. The only clear effect that could be ascribed to the nonalcoholic components in wine was that the LDL-cholesterol was reduced in the long term, although aortic cholesterol was not. PMID- 11383691 TI - Wax ester biosynthesis in the liver of myctophid fishes. AB - The biosynthetic properties of wax esters in the liver were compared between two types of myctophid fishes having different body lipid composition, i.e., three triglyceride-rich species (Lampanyctus jordani, Diaphus theta, and Symbolophorus californiensis) and three wax ester-rich species (L. regalis, Stenobracius nannochir, and Stenobracius leucopsarus). n-Heptadecanol (17:0-ALC) and/or 10-cis heptadecenoic acid (17:1-ACID) was incubated with liver homogenate of the six myctophid fishes and with co-factors such as NADPH and ATP for 2 to 5 h. Considerable amounts of wax esters with odd-numbered fatty acids and/or alcohols were produced in the liver homogenate of the wax ester-rich species. Stenobracius nannochir and L. regalis, which exclusively contained wax esters as neutral lipids, showed the highest activity of wax ester synthesis, followed by S. leucopsarus, which contained triglyceride as the minor constituent. Only trace amounts at most of odd-numbered fatty acids and alcohols were incorporated into the wax esters after incubation with the liver homogenates of the triglyceride rich fishes. Active interchange between the fatty acids and the alcohols occurred during wax ester biosynthesis in the wax ester-rich fishes. The chain elongation and shortening of acyl moieties were also observed during incubation. These results suggested that the deposition of lipids in myctophid fishes is mainly due to their biosynthetic activities. PMID- 11383692 TI - Water-soluble organosulfur compounds of garlic inhibit fatty acid and triglyceride syntheses in cultured rat hepatocytes. AB - The putative hypolipidemic effect of garlic remains controversial. To gain further insight into the effect of garlic on lipid metabolism, the present study determined the inhibitory effects of water-soluble organosulfur compounds present in garlic on triglyceride (TG) and fatty acid synthesis in cultured rat hepatocytes. When incubated at 0.05 to 4.0 mmol/L with cultured hepatocytes, S allyl cysteine (SAC) and S-propyl cysteine (SPC) decreased [2-14C]acetate incorporation into triglyceride in a concentration-dependent fashion achieving a maximal inhibition at 4.0 mmol/L of 43 and 51%, respectively. The rate of [2 14C]acetate incorporation into phosphlipids was depressed to a similar extent by SAC and SPC. SPC, SAC, S-ethyl cysteine (SEC), and gamma-glutamyl-S-methyl cysteine decreased [2-14C]acetate incorporation into fatty acid synthesis by 81, 59, 35, and 40%, respectively, at 2.0-4.0 mmol/L concentrations. Alliin, gamma glutamyl-S-allyl cysteine, gamma-glutamyl-S-propyl cysteine S-allyl-N-acetyl cysteine, S-allylsulfonyl alanine, and S-methyl cysteine had no effect on fatty acid synthesis. The activities of lipogenic enzymes, fatty acid synthase (FAS), and glucose-6-phosphate dehydrogenase (G6PDH) were measured in cultured hepatocytes treated with the inhibitors. The activity of FAS in cells treated with 4.0 mmol/L SAC and SPC, respectively, was 32 and 27% lower than that of nontreated cells. Neither SAC nor SPC affected G6PDH activity. The results indicate that SAC, SEC, and SPC inhibit lipid biosynthesis in cultured rat hepatocytes, and further suggest that these S-alk(en)yl cysteines of garlic impair triglyceride synthesis in part due to decreased de novo fatty acid synthesis resulting from inhibition on FAS. Whether tissue concentrations of active garlic components can achieve levels required to inhibit TG synthesis in vivo warrants further investigation. PMID- 11383693 TI - Regulation by long-chain fatty acids of the expression of cholesteryl ester transfer protein in HepG2 cells. AB - Cholesteryl ester transfer protein (CETP) is an important determinant of lipoprotein function, especially high density lipoprotein (HDL) metabolism, and contributes to the regulation of plasma HDL levels. Since saturated and polyunsaturated fatty acids (FA) appear to influence the CETP activity differently, we decided to investigate the effects of FA on the expression of CETP mRNA in HepG2 cells using an RNA blot hybridization analysis. Long-chain FA (>18 carbons) at a 0.5 mM concentration were added to the medium and incubated with cells for 48 h at 37 degrees C under 5% CO2. After treatment with 0.5 mM arachidonic (AA), eicosapentaenoic (EPA), and docosahexaenoic acid (DHA), the levels of CETP mRNA were less than 50% of the control levels (AA, P = 0.0005; EPA, P < 0.01; DHA, P < 0.0001), with a corresponding significant decrease in the CETP mass. These results suggest that FA regulate the gene expression of CETP in HepG2 and this effect is dependent upon the degree of unsaturation of the acyl carbon chain in FA. PMID- 11383694 TI - Modulation of prostaglandin H synthase activity by conjugated linoleic acid (CLA) and specific CLA isomers. AB - Conjugated linoleic acid (CLA) has been shown to inhibit tumorigenesis in animal models and is cytostatic to numerous cell lines in vitro. However, the mechanism of action is unknown. In the current study, we determined the effects of CLA and specific isomers of CLA on the rate of oxygenation of arachidonic acid by prostaglandin H synthase (PGHS) in ram seminal vesicle microsomes. The enzyme was incubated with 0.1 to 100 microM CLA or specific isomers of CLA for 2 min prior to the addition of 44 to 176 microM arachidonate. The isomers tested were 9(E),11(E) CLA; 9(Z),11(E) CLA; 9(Z),11(Z) CLA, and 10(E),12(2) CLA. For a positive inhibitor control, flurbiprofen was used at 0.75 to 2.50 microM. Enzyme activity was assessed by measuring the rate of oxygen consumption. Inclusion of CLA or specific isomers of CLA in the incubation mixtures inhibits PGHS. The efficacy differs for each isomer, with the 9(Z),11 (E) CLA isomer being the most effective and the 9(Z),11 (Z) CLA isomer being the least effective inhibitor among the four CLA isomers tested. The Ki values obtained by Dixon replots range from 18.7 microM for the most effective isomer, 9(Z),11 (E) CLA, to 105.3 microM for the least effective isomer, 9(2),11(2) CLA. The Ki value for flurbiprofen with ram seminal vesicle microsomes was 0.33 microM. As the concentration of arachidonate was increased, the CLA-dependent inhibition of PGHS decreased, suggesting competitive inhibition. The results of this study demonstrate the potential of CLA and specific isomers of CLA to modulate prostaglandin biosynthesis. PMID- 11383695 TI - Hen egg yolk and white contain high amounts of lysophosphatidic acids, growth factor-like lipids: distinct molecular species compositions. AB - Hen egg yolk and white were found to contain high amounts of lysophosphatidic acid (acyl LPA) in addition to small amounts of lysoplasmanic acid (alkyl LPA). The levels of acyl LPA in hen egg yolk (44.23 nmol/g tissue) and white (8.81 nmol/g tissue) were on the same order as or higher than the levels of acyl LPA known to be required to elicit biological responses in various animal tissues. Noticeably, there is a marked difference between the fatty acid composition of egg yolk acyl LPA and of egg white acyl LPA; egg yolk acyl LPA predominantly contains saturated fatty acids as the acyl moiety, whereas egg white acyl LPA primarily contains polyunsaturated fatty acids. We found that the level of acyl LPA, especially polyunsaturated fatty acid-containing acyl LPA, in egg white was augmented markedly during the incubation at 37 degrees C, while there was no change in egg yolk. We confirmed that egg white contains both the substrate, i.e., polyunsaturated fatty acid-containing lysophosphatidylcholine (LPC), and the enzyme activity catalyzing the hydrolysis of polyunsaturated fatty acid containing LPC to the corresponding acyl LPA. Egg yolk LPA and egg white LPA may play separate physiological roles in the development, differentiation, and growth of embryos. PMID- 11383696 TI - Fullerene lipids: synthesis of novel nitrogen-bridged. AB - Reactions of methyl 6-azido-hexanoate, 8-azidooctanoate, and 12-azido-dodecanoate with [60]fullerene (1) gave the corresponding aza-[60]fullerene ester derivatives (2a-2c, 22-35% based on the amount of [60]fullerene reacted). The nitrogen atom is bonded to the [60]fullerene cage to yield a "[5,6]-open" type aza substructure. This was confirmed by the appearance of 30-31 sp2 signals at deltac 133-147 in the carbon nuclear magnetic resonance spectra. Reaction of methyl 11 azido-7-undecynoate with [60]fullerene furnished a mixture of aza-[60]fullerene (2d, 53%) and aziridine-[60]fullerene (2e, 38%) ester derivatives. Compound 2e was identified as the "[6,6]-closed" type aziridine-[60]fullerene derivative, which displayed 10 sp2 signals in the region deltac 140-145 and one signal at deltac 85.05 for the sp3 carbons of the cage. Refluxing a solution of compound 2d in toluene for 50 h gave about 50% yield of compound 2e, but not vice versa. PMID- 11383697 TI - A new diacylgalactolipid containing 4Z-16:1 from the marine cyanobacterium Oscillatoria sp. AB - A new diacylgalactolipid was isolated from the marine cyanobacterium Oscillatoria sp., and the structure was elucidated as (2S)-3-O-beta-D-galactopyranosyl-1-O (9Z,12Z-oc-tadecadienoyl)-2-O-(4Z-hexadecenoyl)glycerol by enzymatic partial hydrolysis using lipase and physicochemical evidence, which included determining the double bond position in the hexadecenoic acid moiety. PMID- 11383698 TI - Triple-bonded unsaturated fatty acids are redox active compounds. AB - Unsaturated fatty acids with triple bonds are used as inhibitors of unsaturated fatty acid metabolism or cytochrome P450 reactions because they are believed to be chemically inert. In this paper we use in vitro cytochrome C reduction to show that two commonly used triple-bonded unsaturated fatty acids are in fact potent electron transfer agents and could affect the multiple cellular systems that are redox-modulated. PMID- 11383699 TI - Precision of a new thermometer for rapid rectal temperature measurement in neonates. AB - The objective of this article is to evaluate the accuracy and reproducibility of a Penguin electronic thermometer compared with a mercury-in-glass thermometer for rectal thermometry in newborns. The mercury-in-glass thermometer is considered the "gold-standard" for rectal thermometry. Unfortunately, accurate measurement with a mercury-in-glass thermometer requires at least 4 minutes. Rectal temperature was obtained in 224 term and near-term infants using a mercury-in glass and a Penguin thermometers. Paired t-test was used to test the difference between the means, and F-test was used to test the difference between the variances of the two instruments. One hundred and sixty-one of the temperature measurements (72%) taken with the Penguin thermometer were within +/-0.2 degrees C and 208 (93%) within +/-0.5 degrees C from the measurements made with the mercury-in-glass thermometer. The differences in means and in variances were not significantly different between the instruments. The Penguin thermometer is an accurate and reproducible tool for measuring rectal temperature in term and near term infants compared with the mercury-in-glass thermometer. PMID- 11383700 TI - An unusual case of diverticulitis complicating pregnancy at 33 weeks' gestation. AB - A 38-year-old para 2 presented with the gradual onset of nausea vomiting and increasing left lower quadrant pain, at 33 weeks' gestation. She was known to have uterine leiomyomas, with ultrasonography depicting an 8-cm intramural fundal leiomyoma. In addition a left lateral nondiscrete 10 x 8-cm mass was depicted at the point of maximum tenderness. Magnetic resonance imaging (MRI) demonstrated diverticulosis of the descending and sigmoid colon. The patient remained afebrile and received repeated doses of intramuscular analgesics and was cleared by the surgical consultant, only to be readmitted with similar symptomatology 24 hours later. Subsequently, following repeat discharge she delivered at 34 weeks' gestation, and developed a small bowel obstruction during the immediate postpartum course. With the continued finding of a left lower abdominal mass and computed tomography findings suggestive of perforated sigmoid diverticulitis and resulting small bowel obstruction, laparotomy was performed. Multiple adhesions and phlegmon sequelae of chronic perforation of the sigmoid were confirmed, and a diverting descending colostomy and Hartman's procedure were performed. We present unusual MRI findings of diverticulitis in the third-trimester and review the literature pertaining to this unusual complication of pregnancy. PMID- 11383701 TI - Transient fetal hypothyroidism due to direct fetal administration of amiodarone for drug resistant fetal tachycardia. AB - Amiodarone, an anti-arrhythmic drug that contains 39% iodine, is rarely known to cause negative effects on fetal thyroid function after gestational exposure, when given orally to a pregnant woman. Two cases of fetal hypothyroidism after gestational exposure to amiodarone by direct fetal intravenous route are described here. PMID- 11383702 TI - From TPN to breast feeding--feeding the premature infant--2000: Part I. Parenteral nutrition. AB - Advancements in medical technology have allowed for the survival of smaller, more premature infants, during the past several years. Infants as immature as 23 weeks gestational age and as small as 500 g, or even less, are now common sights in Neonatal Intensive Care Units throughout the United States. These infants present a unique challenge to neonatologists and registered dietitians, who must provide them with the nutrients needed for normal growth and development. With mounting evidence of a direct link between early nutrition and later outcome, the need for a better understanding of fetal and neonatal nutrition is critical. From admission to discharge, most infants will transition from a dependence on parenteral feeding, to a dependence on tube feeding, and finally, the ability to orally feed. In the early days of postnatal life, decisions must be made about when to initiate TPN, what type of intravenous access will be required, and to what extent and at what rate parenteral nutrients should be advanced. Similar decisions are made about the introduction and advancement of enteral nutrition. The following is a review of the available literature on the parenteral feeding of premature infants, with an emphasis on how our practice in the year 2000 was evolved over time. A similar review of enteral feeding practices will be presented in a future installment. PMID- 11383703 TI - The myth of transient hypertension: descriptor or disease process? AB - The objective of this study is to describe the incidence of transient hypertension and to evaluate if transient hypertension is associated with increased maternal or fetal morbidity as compared to other hypertensive disorders of pregnancy and normotensive controls. Data were collected from all deliveries at the University of Mississippi Medical Center from July 1, 1996 through January 1, 1997. Patients were grouped according to ACOG criteria for pregnancy induced hypertension. Specific maternal and fetal morbidities were compared among the groups and controls. There were 1489 deliveries during the study period. Nearly 30% of patients met the criteria for transient hypertension. There were no significant differences between patients with transient hypertension and controls in regard to maternal and fetal outcomes. Transient hypertension occurs more often than expected, however, it appears to be of limited clinical significance. PMID- 11383704 TI - Late-onset cystic periventricular leukomalacia in premature infants: a threat until term. AB - The purposes of this study are (1) to describe a "late-onset" form of cystic periventricular leukomalacia eventually appearing in premature infants whose neurological assessments were normal in the first month of life; (2) to retrospectively evaluate its incidence among a large population of premature infants; (3) to suggest that a few unexpected complications of prematurity may trigger the development of white matter damage, even several weeks after birth. Retrospective study in a population of 1452 surviving infants after 5 days born before 33 weeks. We identified 10 cases of late-onset cystic periventricular leukomalacia appearing beyond the first 5 weeks of life. In 8 cases, an intercurrent event associated with a systemic inflammatory response preceded the appearance of cysts: necrotizing enterocolitis (n = 5), septicemia (n = 2 cases), strangulated inguinal hernia in one infant. Neurological surveillance should be repeated until discharge in very preterm infants, especially after the occurrence of an intercurrent complication coming along with a systemic inflammatory response. PMID- 11383705 TI - Neonatal-perinatal risk factors for the development of persistent pulmonary hypertension of the newborn in preterm newborns. AB - There is a long-held belief that preterm newborns lack sufficient arteriolar musculature to maintain a prolonged elevated pulmonary vascular resistance (PVR) after birth. Net ductal flow is thought to be minimal, with the developing pulmonary circulation incapable of significant vasoconstriction. We identified retrospectively 15 premature newborns over a 10-year period weighing < or = 1500 g and with a gestational age of < or = 30 weeks with documented persistent pulmonary hypertension of the newborn (PPHN) in the first 24 hours after birth. We matched 36 newborns of similar weight and gestation with no clinical evidence of shunting. The control group weaned to an FiO2 < or = 0.50 by 12 hours after birth. Despite similar gestational ages, the PPHN group (n = 15) had significantly higher birth weights than the control group (n = 36). The duration of ruptured membranes, maternal tobacco use, and use of antenatal steroids were significantly higher in the PPHN group. We speculate that these three factors might act in a synergistic relationship with which to accelerate pulmonary vascular smooth muscle development in premature newborns. PMID- 11383706 TI - Nucleated red blood cell counts in term neonates with umbilical artery pH < or = 7.00. AB - The purpose of this study was to determine whether nucleated red blood cell (NRBC) counts are elevated in term neonates who have severe fetal acidemia at birth. The neonatal NRBC counts of term (gestational age > or = 37 weeks) neonates with pathological acidemia were compared with those from control neonates who met the following criteria: gestational age > or = 37 weeks, birth weight > or = 2800 g, umbilical artery pH > or = 7.25, and a 5-minute APGAR > 7. Pathological acidemia was defined as an umbilical artery pH < or = 7.0 and a base excess > -12 mEq/L. Twenty-six neonates met all inclusion criteria and were compared to 78 controls. The mean NRBC/100 WBC was 11.9 +/- 13.5 (range 0 to 45) for acidemic neonates compared to 3.9 +/- 2.9 NRBC/100 WBC (range 0 to 11) for control neonates [p <0.001]. Our findings suggest that the onset of hypoxia ischemia in pregnancies complicated by severe fetal acidemia often begins prior to the intrapartum period. PMID- 11383707 TI - Spontaneous umbilical cord hematoma: a rare cause of acute fetal distress. AB - Spontaneous umbilical cord hematoma is very rare. We present a newborn who was discovered to have a spontaneous fetal umbilical cord hematoma. A nonreassuring fetal heart tracing complicated the first and second stages of labor. The only pathological finding was a short umbilical cord, which could have contributed to the rupture of fetal vessels. PMID- 11383708 TI - Electrically evoked release of glutamate in rat hippocampal slices: effects of various drugs and fimbria-fornix lesions. AB - A model of electrically evoked release of glutamate from rat hippocampus was developed and used to detect possible changes induced by lesions of hippocampal afferences. Neuronal glutamate in hippocampal slices was labelled by preincubation with [3H]glutamine. The slices were then superfused with physiological medium in the presence of the glutamate uptake inhibitor L transpyrrolidine-2,4-dicarboxylic acid (100 microM or 3 microM) and stimulated twice electrically (S1, S2: 240 pulses, 3 Hz, 2 ms, 26-30 mA); various drugs were added before S2. In order to determine the basal and evoked outflow of [3H]glutamate only, the mixture of 3H-labelled compounds (glutamine, glutamate and GABA) was separated by ion exchange chromatography in superfusate fractions and slices. The electrically evoked overflow of [3H]glutamate was largely Ca2+ dependent and tetrodotoxin-sensitive and hence represented action potential induced exocytotic release of [3H]glutamate. Evoked [3H]glutamate release was significantly increased by the adenosine A1 receptor antagonist 8-cyclopentyl-1,3 dipropylxanthine (DPCPX, 0.1 microM), suggesting the presence of endogenous inhibitory adenosine, and reduced by the A1 receptor agonist N6 cyclopentyladenosine (1 microM, antagonized by DPCPX, 0.1 microM). There was no evidence for a cholinergic, serotonergic, or adrenergic modulation of the evoked release of [3H]glutamate: the corresponding selective agonists (or antagonists) were ineffective. After aspirative lesions of the septohippocampal pathways the hippocampal noradrenaline content was markedly increased, whereas cholinergic and serotonergic markers were reduced. The evoked release of [3H]glutamate in hippocampal slices of lesioned rats was significantly increased by a mechanism which still has to be determined, but which is not related to alterations in A1 receptor function. It is concluded that the present model was able to detect lesion-induced differences in electrically evoked release of [3H]glutamate, but the relationship of these differences to changes of noradrenergic, cholinergic or serotonergic hippocampal innervations remains to be established. PMID- 11383709 TI - Opposite effects of antidepressants and corticosterone on the sensitivity of hippocampal CA1 neurons to 5-HT1A and 5-HT4 receptor activation. AB - Using extracellular and intracellular ex vivo recording techniques we studied changes in the reactivity of hippocampal pyramidal CA1 neurons to serotonin (5 HT) and to the 5-HT1A- and 5-HT4 receptor agonists (+/-)-2-dipropylamino-8 hydroxy- 1,2,3 ,4-tetrahydronaphthalene hydrobromide (8-OH-DPAT) and zacopride, respectively, evoked by repeated electroconvulsive shock (ECS), imipramine and corticosterone treatments. Rats were subjected to ECS for 1 or 10 days, treated with imipramine for 1, 7, 14 or 21 days (10 mg/kg p.o., twice daily) and with corticosterone for 7 days (10 mg/kg s.c., twice daily). Hippocampal slices were prepared 2 days after the last treatment. Activation of 5-HT1A receptors decreased the amplitude of population spikes evoked by stimulation of the Schaffer/collateral-commissural pathway and hyperpolarized CA1 cells. Activation of 5-HT4 receptors increased the population spike amplitude and decreased the amplitude of slow afterhyperpolarization. Both repeated ECS and imipramine enhanced the effects related to 5-HT1A receptor activation and attenuated the effects of 5-HT4 receptor activation. The action of imipramine was significant after a 7-day treatment and reached a maximum after 14 daily applications, remaining at the same level in a group of animals treated for 21 days. Repeated corticosterone attenuated the inhibitory effect of 5-HT and 8-OH-DPAT on the population spike amplitude and enhanced the increase in population spike amplitude induced by zacopride. These findings indicate that antidepressant treatments and repeated corticosterone have opposite effects on hippocampal responsiveness to 5-HT1A and 5-HT4 receptor activation. In consequence, antidepressants enhance, whereas corticosterone reduces the 5-HT-mediated inhibition of hippocampal CA1 cells, which may be relevant to the antidepressant and pro-depressant effects of either treatment, respectively. PMID- 11383710 TI - Effect of myelopoietic and pleiotropic cytokines on colony formation by blast cells of children with acute lymphoblastic leukemia. AB - The aim of this study was to see whether pleiotropic or myeloid hematopoietic growth factors, which do not stimulate normal lymphoid cells, can induce proliferation of blast cells of the acute lymphoid leukemia (ALL) of childhood. Bone marrow cells of 13 children with untreated ALL (nine common ALL, two myeloid antigen positive ALL and two early T-cell ALL) formed colonies of leukemic blast cells in primary methylcellulose cultures. Spontaneous growth was observed in three of 13 cases, whereas phytohemagglutinin-stimulated leukocyte conditioned medium (PHA-LCM), a conventional source of various natural human cytokines, induced colony formation in ten of 13 cases. A similar rate of responsiveness was seen with recombinant human granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF) and stem cell factor (SCF); a combination of these three cytokines induced colony formation in all cases studied. The effect of these growth factors on colony formation seemed to be dose-dependent in some cases. Of the stimuli studied, GM-CSF induced the smallest number of colonies, whereas the effects of G-CSF, SCF and PHA-LCM were similar in this respect. Combination of cytokines proved to be even more efficient in inducing clonal proliferation of leukemic lymphoblasts. In double combinations, G-CSF and GM-CSF as well as G-CSF and SCF were able to potentiate each other's effects. Triple combination of these cytokines mediated the most potent growth stimulus. Our results demonstrate that myeloid and pleiotropic cytokines are able to stimulate clonal proliferation of pediatric leukemic lymphoblasts. This may present a potential hazard to children with ALL while on adjuvant therapy with hematopoietic growth factors. In vitro colony assays performed prior to or in parallel with the administration of hematopoietic growth factors to ALL patients may help to forecast their possible effects on leukemic cells in vivo. PMID- 11383711 TI - Differential effects of eprosartan and losartan at prejunctional angiotensin II receptors. AB - A comparison was made of the influence of losartan and eprosartan on angiotensin II effects at pre- and postjunctional receptors of the canine pulmonary artery and at prejunctional receptors of the rat left ventricle. To study postjunctional contractile responses to angiotensin II, non-cumulative concentration-response curves were determined; to study prejunctional effects of angiotensin II, the tissues were preincubated with [3H]noradrenaline and then superfused and electrically stimulated (1 Hz, 2 ms, 50 mA, 5 min). Postjunctionally, both losartan and eprosartan caused a parallel shift of the concentration-response curve of angiotensin II to the right (pKd of 8.15 and 8.28, respectively). At the prejunctional level, while eprosartan, in concentrations similar to those which were effective postjunctionally (30-100 nM), antagonized the facilitatory effect on noradrenaline release in both the dog pulmonary artery and the rat ventricle, losartan was ineffective in concentrations up to 1 microM. It is concluded that pre-junctional receptors for angiotensin II in the canine pulmonary artery and in the rat left ventricle are different from postjunctional receptors of the canine pulmonary artery. It is proposed that the prejunctional receptors of these tissues are atypical AT1 or "AT1B-like" receptors. PMID- 11383712 TI - Effects of modulating Ca2+ entry and activating prejunctional receptors on facilitation of excitatory junction potentials in the guinea-pig vas deferens in vitro. AB - This study investigated the effects of changing the extracellular Ca2+ concentration on facilitation of excitatory junction potential (EJP) amplitude during trains of 20 stimuli at 1 Hz at sympathetic neuroeffector junctions in the guinea-pig vas deferens. These effects were compared with those of Ca2+ channel blockers and agents which act at prejunctional receptors to increase or decrease neurotransmitter release. In these experiments, alpha-adrenoceptor-mediated autoinhibition of neurotransmitter release was blocked by the alpha-adrenoceptor antagonist, phentolamine (1 microM). Varying the extracellular Ca2+ concentration (0.75-6 mM) changed the amplitude of EJPs without affecting the maximum level of facilitation during the trains of stimuli. Reductions in Ca2+ concentration (from 2 mM) were associated with a slowing in the rate of development of facilitation. The Ca2+ channel antagonists, Cd2+ (2 microM and 5 microM) and omega-conotoxin GVIA (10 nM), and agents which act at prejunctional receptors to reduce neurotransmitter release, adenosine (100 microM and 1,000 microM) and prostaglandin E2 (PGE2; 0.1 nM and 1 nM), produced similar effects to those of lowering the extracellular Ca2+ concentration. Raising the extracellular Ca2+ concentration (from 2 mM) increased the rate of development of facilitation. Angiotensin II (AII; 0.5 microM) produced similar effects to raising extracellular Ca2+. However, isoprenaline (1 microM), while increasing EJP amplitude, reduced the maximum level of facilitation and was without effect on the rate of development of facilitation. In the guinea-pig vas deferens EJPs are produced by neurally released ATP. Thus, the findings support the idea that adenosine, PGE2 and AII change ATP release by modifying Ca2+ entry into the nerve terminal. However, the effects of isoprenaline may not solely be accounted for by modifications in Ca2+ entry. PMID- 11383713 TI - Analysis of ligand activation of alpha 2-adrenoceptor subtypes under conditions of equal G alpha protein stoichiometry. AB - Variations in the measurement of ligand's intrinsic activity between receptor subtypes is a common consequence of unequal receptor:G protein density ratios. We have investigated ligand activation at the alpha2-adrenoceptor (alpha2-AR) subtypes under defined expression conditions of one receptor molecule for one Galpha protein molecule using fusion proteins. Fusion between either a wt alpha2C AR or a mutant Thr382Lys alpha2C AR and a chimeric Galphaq/il protein displayed robust, transient (-)-adrenaline-mediated Ca2+ responses with similar potencies (pEC50: 7.78 and 7.66) and kinetic properties. A comparison of the intrinsic activities of alpha2 AR agonists found d-medetomidine to be the only compound with an efficacy similar to that of (-)-adrenaline. The Ca2+ responses as mediated by UK 14304, oxymetazoline and clonidine became more potent and efficacious at the Thr381Lys alpha2C AR, whereas the response as mediated by talipexole displayed a higher potency with an unaltered maximal response. Whereas only small differences in ligand's intrinsic activities between the wt alpha2A, alpha2B and alpha2C AR fusion proteins were observed with most ligands, oxymetazoline was virtually silent at the alpha2A AR while active as a partial and apparently full agonist at the alpha2C AR and alpha2B AR, respectively. The mutant alpha2 AR subtypes could be differentiated using the apparent positive efficacy of ligands that used to be defined as antagonists. The following rank order of maximal responses was observed for the Thr381Lys alpha2C AR: idazoxan approximately equals SKF 86466 > atipamezole >> dexefaroxan; Thr373Lys alpha2A AR: SKF 86466 > idazoxan = atipamezole > dexefaroxan; and Thr370Lys alpha2B AR: atipamezole > idazoxan dexefaroxan. RX 811059 (10 microM) was the only compound to be completely silent at both the wt and mutant alpha2 AR subtypes. In conclusion, silent alpha2 AR ligands are probably rare in these specified alpha2 AR systems. Most antagonists may actually possess partial agonist properties at the alpha2 AR subtypes, which are facilitated by the same mutation in the distal portion of their third intracellular loop. PMID- 11383714 TI - Modulation of K+-evoked [3H]-noradrenaline release from rat and human brain slices by gabapentin: involvement of KATP channels. AB - To elucidate the mechanism of action of the anticonvulsant gabapentin (GBP), we compared its effects on K+-evoked [3H]-noradrenaline ([3H]-NA) release from rat hippocampal and human neocortical slices with those of the KATP channel opener pinacidil and the Na+ channel blockers phenytoin, carbamazepine and lamotrigine. Rat hippocampal and human neocortical slices were loaded with [3H]-NA and superfused. [3H]-NA release was evoked by increasing the extracellular [K+] from 3 to 15 mM. GBP decreased [3H]-NA release from rat hippocampal with a pIC50 of 5.59 and a maximum inhibition of 44%. Concentration-dependent inhibition was also seen in human neocortical slices (39% inhibition with 100 microM GBP). These inhibitory effects were antagonized by the KATP channel antagonist glibenclamide, yielding a pA2 of 7.50 in the rat. The KATP channel opener pinacidil (10 microM), like GBP, decreased [3H]-NA release from rat hippocampal slices by 27% and this effect was also antagonized by glibenclamide. In human neocortical slices the inhibition by pinacidil (10 microM) was 31%. Although phenytoin (10 microM), carbamazepine (100 microM) and lamotrigine (10 microM) also decreased [3H]-NA release (by 25%, 57% and 22%, respectively), glibenclamide did not antagonize the effects of these classical Na+ channel blockers. These findings suggest that GBP inhibits K+-evoked [3H]-NA release through activation of KATP channels. To establish whether the KATP channels under investigation were located on noradrenergic nerve terminals or on other neuronal elements, the effects of GBP were compared in the absence and in the presence of tetrodotoxin (TTX 0.32 microM) throughout superfusion. Since the functional elimination of the perikarya of interneurons by TTX reduced the inhibitory effect of GBP, the KATP channels mediating the effect of GBP may be located on nerve terminals, probably on both noradrenergic and glutamatergic nerve endings. PMID- 11383715 TI - Mechanical response to electrical field stimulation of rat, guinea-pig, monkey and human detrusor muscle: a comparative study. AB - The aim of the present investigation was to compare mechanical responses to electrical field stimulation (EFS), as well as cholinergic and non-adrenergic, noncholinergic (NANC) neurotransmission in guinea-pig, rat, monkey and human detrusor muscle strips. Responses to EFS (0.05, 0.5 and 1 ms pulse duration, 50 V, 1-15 Hz) of guinea-pig, rat, monkey and human detrusor muscle strips were recorded isometrically before and after blockade of muscarinic receptors and/or P2-purinoreceptors, as well as after desensitisation of P2-purinoceptors or blockade of the nerve impulse propagation. Single pulses of 0.05 ms duration elicited responses, in either guinea-pig or rat detrusor strips, which were abolished by tetrodotoxin (TTX), thus suggesting their neurogenic nature. In monkey and human detrusor strips, however, the same single pulses were not sufficient to generate contractile responses. The response of either rat or guinea-pig strips to single pulses of 0.5 ms and 1 ms duration was mainly myogenic in nature. While in rat and guinea-pig strips the neurogenic response was only partly reduced in the presence of atropine, in monkey and human strips it was abolished. In the presence of atropine, while suramin only partially reduced the EFS response either in rat or guinea-pig detrusor strips, a complete alpha,beta-methyleneATP-sensitive response was evident in guinea-pig detrusor strips. This suggests the involvement of other transmitter(s) beyond ATP in the NANC response of rat detrusor strips. PMID- 11383716 TI - Effects of Ro 64-6198 in nociceptin/orphanin FQ-sensitive isolated tissues. AB - The pharmacological profile of the non-peptide OP4 receptor (ORL1, LC132) agonist, Ro 64-6198, was investigated, in three electrically stimulated nociceptin/orphanin FQ (NC)-sensitive preparations, namely the mouse and rat vas deferens and the guinea pig ileum. Ro 64-6198 mimicked the inhibitory effect of NC in the three preparations, while showing slow kinetics of action and a slowly reversible effect compared to the fast and immediately and completely reversible effect of the natural peptide. Ro 64-6198 displayed similar pEC50 and Emax values as NC in the mouse and rat vas deferens while it was 100-fold less potent but more efficacious (higher Emax) than NC in the guinea pig ileum. In the rat vas deferens the effects of Ro 64-6198 were antagonised by [Nphe1]NC(1-13)NH2 and J 113397 with pKB values (6.30 and 8.05, respectively) similar to those obtained against NC (6.20 and 7.77, respectively). Naloxone (1 microM) was inactive. In the guinea pig ileum a clear shift of the concentration response curve to Ro 64 6198 was obtained only using a cocktail of antagonists (naloxone + [Nphe1]NC(1 13)NH2 or naloxone + J-113397). In the mouse vas deferens the antagonists were inactive against Ro 64-6198 either when tested alone or in combination. Therefore, Ro 64-6198 behaved as a selective OP4 receptor agonist only in the rat tissue. These results suggest a physiological heterogeneity in OP4 receptors across tissues and species and may explain why, when tested in vivo, Ro 64-6198 mimics the potent anxiolytic effect of NC better in the rat than in the mouse. PMID- 11383717 TI - Effects of smoking cessation and nicotine substitution on systemic eicosanoid production in man. AB - This study investigated the effects of smoking cessation with and without nicotine substitution on the excretion of major urinary metabolites of thromboxane A2 and prostacyclin, 11-dehydrothromboxane B2 and 2,3-dinor-6 ketoprostaglandin F1alpha, respectively, as well as on the excretion of leukotriene E4 in man. Urine samples were obtained from 20 healthy non-smoking controls and from 60 healthy smoking volunteers before, and 3, 7 and 14 days after smoking cessation. Fifteen smokers quit smoking without nicotine substitution, 15 used nicotine chewing gum and 30 used nicotine patches as a substitution therapy. Urinary thiocyanate as well as cotinine and trans-3' hydroxycotinine excretions were used as compliance and nicotine substitution indicators. 11-Dehydrothromboxane B2, 2,3-dinor-6-ketoprostaglandin F1alpha and leukotriene E4 excretion was about two, three and five times higher in smokers than in controls, respectively. Three days after smoking cessation without nicotine substitution, 11-dehydrothromboxane B2 and 2,3-dinor-6-ketoprostaglandin F1alpha levels were lowered to 75% (P<0.01) and 80% (P<0.05) of the initial values, and after 14 days to 50% (P<0.01) and 60% (P<0.05), respectively. In 3 days leukotriene E4 excretion was dropped to 70% of the initial value (P<0.05), but no further decrease was observed during the study. In individuals using nicotine chewing gum or nicotine patches no significant changes were observed in the analytes during the 2-week follow-up. The increased systemic eicosanoid synthesis observed in smokers may be involved in the harmful cardiovascular effects of smoking. The fact that eicosanoid production remains at pre-cessation level in volunteers who quit smoking but use nicotine substitution may be involved in the risk of cardiovascular complications reported during nicotine replacement therapy. PMID- 11383718 TI - Loss of locomotor sensitisation in response to morphine in D1 receptor deficient mice. AB - Mice lacking D1 receptors were used to study the role of these receptors in morphine-induced antinociception and locomotor sensitisation. In the hot-plate test D1 receptor deficient (-/-) and wild-type (+/+) mice showed similar reaction times under basal conditions. A single injection of 1.25 mg/kg and 2.5 mg/kg morphine resulted in a stronger antinociceptive response in D1 receptor deficient mice than in wild-type animals. Tolerance to the analgesic effect did not develop in both groups of animals when 12.5 mg/kg morphine was chronically applied twice daily for 13 days. There was no change in basal locomotor activity between saline injected wild-type and D1 receptor deficient mice. After chronic treatment wild type mice showed a continuous increase in locomotor activity, indicating the development of sensitisation. In contrast, a subchronic administration of morphine did not change locomotor activity in mutant mice. The lack of the development of locomotor sensitisation in D1 deficient mice was associated with reduced levels of immunoreactive mu opioid receptors in dorsal striatal patches as compared to wild-type mice. In contrast, no change in the distribution of immunoreactive mu receptors could be detected in areas related to pain pathways such as the spinal cord. Taken together, these results suggest an involvement of D1 receptors in morphine-induced locomotor activity and analgesia. PMID- 11383719 TI - Glutamatergic-cholinergic synergistic interaction in the pontine reticular formation. Effects on catalepsy. AB - Microinjections of small amounts of the cholinergic receptor agonists carbachol and nicotine into the pontine reticular formation (PRF) of rats were shown to induce catalepsy. Catalepsy was used in this work as an experimental model for studying the interactions between cholinergic mechanisms and excitatory amino acid mechanisms in the PRF. The excitatory amino acid (EAA) receptor agonists glutamate, NMDA, kainate and AMPA were microinjected in subcataleptic doses before carbachol in the same location into the PRF. All the EAA receptor agonists injected induced a significant potentiation of the cataleptogenic effect of carbachol. The NMDA receptor antagonist MK-801 and the non-NMDA receptor antagonist DNQX microinjected in picomol doses before the EAA receptor agonists attenuated their potentiating effect. These results support the suggestion that EAA neuronal mechanisms contribute synergistically with the cholinergic mechanisms to the PRF neuronal interactions involved in the generation of catalepsy. Similar synergistic interactions might be active in the generation of other pontine behavioral manifestations like REM sleep. PMID- 11383720 TI - Acetylcholine-induced positive inotropy mediated by prostaglandin released from endocardial endothelium in mouse left atrium. AB - The possible involvement of the endocardial endothelium in the positive inotropic response of the mouse left atrium to acetylcholine was examined pharmacologically. In mouse left atria, acetylcholine produced a biphasic inotropic response: a transient decrease in contractile force followed by a late increase. The positive response was not affected by the presence of phentolamine and propranolol, but was almost abolished by pretreatment of the preparation with 1% Triton X-100, which denudes the endocardium of its endothelium. Nordihydroguaiaretic acid, NG-nitro-L-arginine, BQ-123 and BQ-788 had no effect on the inotropic responses to acetylcholine, but indomethacin completely abolished the positive response. Prostaglandins and their analogues had a positive inotropic effect with a potency order PGF2alpha>PGD2>PGE2>U46619, whereas beraprost had no effect. Neither Triton X-100 pretreatment nor the presence of indomethacin affected the positive inotropic effect of PGF2alpha. Acetylcholine and PGF2alpha prolonged the action potential duration similarly. These results suggest that the acetylcholine-induced positive inotropic response in mouse left atria is mediated by prostaglandin released from the endocardial endothelium. PMID- 11383721 TI - Current multimodality management of infectious intracranial aneurysms. AB - OBJECTIVE: To implement an algorithm for and assess multimodality (medical, endovascular, and microsurgical) treatment of patients with infectious intracranial aneurysms. METHODS: Twenty patients with 27 infectious aneurysms were treated during a 10-year period. Bacterial endocarditis was the most common cause (65%). Most aneurysms presented with rupture (75%), and the middle cerebral artery was the most common location (70%). RESULTS: Five patients were treated endovascularly, with direct coiling for three patients and parent artery occlusion for two patients. Ten patients (15 aneurysms) were treated surgically, with 6 aneurysms being trapped/resected, 2 trapped/bypassed, 4 clipped, and 3 wrapped. Five patients were treated medically. Treatment-associated neurological morbidity was observed for two patients (10%), and two patients died (10%). Good outcomes were observed for 16 patients (80%). CONCLUSION: Factors that guide management decisions for these patients include aneurysm rupture, hematomas with increased intracranial pressure, and the eloquence of brain tissue supplied by the parent artery. Patients with unruptured infectious aneurysms are initially treated medically, with antibiotics and serial angiography. Patients with ruptured aneurysms that are not associated with hematomas and that do not involve eloquent vascular territory are treated endovascularly. Patients with ruptured aneurysms are treated surgically when there is a hematoma or the risk of ischemic complications in eloquent territory. Therefore, endovascular therapy is the first option for patients in stable condition with ruptured aneurysms; surgical therapy is the first option for patients in unstable condition with ruptured aneurysms and the second option for patients in stable condition who experience failure of endovascular therapy. Medically treated patients with enlarging or dynamic unruptured aneurysms also require direct surgical or endovascular intervention. Favorable patient outcomes can be achieved with this multimodality management. PMID- 11383722 TI - Transluminal stent-assisted angiplasty of the intracranial vertebrobasilar system for medically refractory, posterior circulation ischemia: early results. AB - OBJECTIVE: Symptomatic vertebrobasilar artery stenosis portends a poor prognosis, even with medical therapy. Surgical intervention is associated with considerable morbidity, and percutaneous angioplasty alone has demonstrated mixed results, with significant complications. Recent advances in stent technology have allowed for a novel treatment of symptomatic, medically refractory, vertebrobasilar artery stenosis. We report on a series of patients with medically refractory, posterior circulation stenosis who were treated with transluminal angioplasty and stenting at two medical centers in the United States. METHODS: A retrospective analysis of data for 11 consecutive patients with symptomatic, medically refractory, intracranial, vertebral or basilar artery stenosis was performed. All patients were treated with percutaneous transluminal angioplasty and stenting. Short-term clinical and angiographic follow-up data were obtained. RESULTS: Among 11 patients who were treated with stent-assisted angioplasty of the basilar or vertebral arteries, there were three periprocedural deaths and one delayed death after a pontine stroke. Other complications included a second pontine infarction, with subsequent residual diplopia. The remaining seven patients (64%) experienced symptom resolution and have resumed their preprocedural activities of daily living. Angiographic follow-up examinations demonstrated good patency of the stented lesions for five of seven survivors (71%); one patient exhibited minimal intrastent intimal hyperplasia, and another patient developed new stenosis proximal to the stent and also developed an aneurysm within the stented portion of the basilar artery. The last patient exhibited 40% narrowing of the treated portion of the vessel lumen. CONCLUSION: Recent advances in stent technology allow negotiation of the proximal posterior circulation vasculature. Although the treatment of vertebrobasilar artery stenosis with angioplasty and stenting is promising, long-term angiographic and clinical follow-up monitoring of a larger patient population is needed. PMID- 11383723 TI - Venous congestion is a major cause of neurological deterioration in spinal arteriovenous malformations. AB - OBJECTIVE: Although venous congestion is considered to be a major cause of progressive myelopathy in patients with spinal dural arteriovenous fistulae (DAVFs), the neurological deterioration in patients with spinal intradural arteriovenous malformations (AVMs) has been attributed to hemorrhage or to vascular steal. To reexamine this theory, we analyzed our own cases of spinal vascular diseases. METHODS: In 24 patients with spinal vascular diseases, those who demonstrated progressive myelopathy with T2 hyperintensity in the spinal cord on magnetic resonance imaging (MRI) were diagnosed as patients with congestive myelopathy. We further examined the clinical courses, MRI findings, and reversibility of these cases. RESULTS: Venous congestion was judged to be a cause of neurological deterioration in 13 patients (7 DAVFs, 6 intradural AVMs). The T2 signals on these patients' MRI scans were located in the center and extended over several levels not corresponding to distribution of ischemia due to arterial steal. Of the patients who were diagnosed with congestive myelopathy, no differences between those with DAVFs and those with intradural AVMs were apparent in terms of clinical manifestations and reversibility. Eight (four DAVFs, four intradural AVMs) of 13 patients experienced neurological improvement after treatment. All patients with poor outcomes had intervals from onset of more than 3 years and showed contrast enhancement of the spinal cord on MRI studies. CONCLUSION: Spinal intradural AVMs as well as spinal DAVFs can be a cause of venous congestive myelopathy. Regardless of its etiology, congestive myelopathy is potentially reversible if properly diagnosed and treated. PMID- 11383724 TI - Radiosurgery for the treatment of recurrent central neurocytomas. AB - OBJECTIVE: Central neurocytomas are benign neoplasms with neuronal differentiation typically located in the lateral ventricles of young adults. Although the treatment of choice is complete surgical excision, patients may experience local recurrence. Adjuvant therapy for patients with residual or recurrent tumor has included reoperation, radiotherapy, or chemotherapy. To avoid the side effects of conventional radiotherapy in young patients, we present a series of patients with clear evidence of tumor progression who were treated with gamma knife radiosurgery. METHODS: Four patients (ages 20-49 yr; mean, 28 yr) who presented with an intraventricular mass on magnetic resonance imaging scans and underwent craniotomy for tumor resection were reviewed retrospectively. Histopathological analysis confirmed central neurocytoma in all cases. Each patient was followed up clinically and radiographically with serial magnetic resonance imaging. When radiographic signs of tumor progression were evident, patients were treated with radiosurgery. RESULTS: Complete radiographic tumor resection was achieved in all patients. There were no major postoperative complications. Local tumor progression was detected on magnetic resonance imaging scans 9 to 25 months after surgery (median, 17.5 mo). All patients achieved complete response to radiosurgery with reduction in tumor size. There have been no complications from radiosurgery. Follow-up ranged from 12 to 28 months (mean, 16.5 mo) after radiosurgery, and from 24 to 84 months (mean, 54.5 mo) after initial presentation. CONCLUSION: Radiosurgery with the gamma knife unit provides safe and effective adjuvant therapy after surgical resection of central neurocytomas. Radiosurgery may eliminate the need for reoperation and avoid the possible long-term side effects from conventional radiotherapy in young patients. PMID- 11383725 TI - Transsphenoidal surgery for acromegaly: endocrinological follow-up of 98 patients. AB - OBJECTIVE: Transsphenoidal surgery is the preferred treatment modality for growth hormone (GH)-secreting pituitary adenomas. In many series, the reported postoperative remission is based mainly on achievement of GH levels less than 2 ng/ml. Strict criteria for insulin-like growth factor I normalization and even lower GH levels (<1 ng/ml) are now suggested to define cure of acromegaly, but the evidence does not yet support such low GH levels in epidemiological follow up. We analyzed our postoperative results in a large cohort of patients with acromegaly. METHODS: Ninety-eight patients harboring GH-secreting adenomas (46 microadenomas and 52 macroadenomas) underwent transsphenoidal surgery between 1990 and 1999. Ninety-one patients were operated for the first time, and 12 patients underwent reoperations because of previous surgical failure (7 had undergone surgery elsewhere previously). Biochemical remission was defined as a repeated fasting or glucose-suppressed GH level of 2 ng/ml or less, and a normal insulin-like growth factor I level. RESULTS: Remission was achieved in 74% of all patients after one operation, including 84% of patients with microadenomas and 64% of patients with macroadenomas. Seventy-three percent of patients with macroadenomas 11 to 20 mm in size achieved remission, as compared with a 20% remission rate for patients with adenomas larger than 20 mm. Patients with preoperative random GH levels lower than 50 ng/ml had a better outcome (85% remission), whereas GH greater than 50 ng/ml was associated with remission in 30% of the patients. Only one of the patients (8%) with postoperative active disease who underwent a second operation achieved remission. Recurrence was rare (one patient), and all failed surgical attempts could be detected during the immediate postoperative evaluation. CONCLUSION: On the basis of strict postoperative GH and insulin-like growth factor I criteria to define remission, our series demonstrates the efficacy of transsphenoidal surgery for acromegalic patients with microadenomas and noninvasive macroadenomas. However, patients with large adenomas (>20 mm) and preoperative GH greater than 50 ng/ml have a poor prognosis and require adjunctive medical or radiation therapy to control GH hypersecretion. PMID- 11383726 TI - Management of hydrocephalus associated with vestibular schwannoma and other cerebellopontine angle tumors. AB - OBJECTIVE: Hydrocephalus (HCP) resulting from cerebellopontine angle (CPA) tumors is not rare. This retrospective study was designed to investigate the incidence of HCP and the clinical presentations, management options, and outcomes of HCP in 284 patients with CPA tumors. METHODS: A retrospective study of 284 consecutive patients with CPA tumors (mostly vestibular schwannomas) treated from 1985 to 1996 at Toronto Western Hospital managed by one surgical team consisting of a neurosurgeon and a neuro-otologist. RESULTS: Thirty-nine patients (13.7%) had radiographic and/or clinical evidence of HCP, 37 preoperatively and 2 postoperatively. Tumor type distribution was 33 vestibular schwannomas, 5 meningiomas, and 1 cavernous hemangioma. Only five patients (12%) had obvious obstruction at the fourth ventricular level. In 36 patients (92%), symptoms were mostly chronic and mild, consistent with normal pressure hydrocephalus. Multivariate analysis confirmed the strong association of tumor size and incidence of HCP (P < .0001). Four patients underwent permanent shunting before microsurgical tumor excision, mainly because of florid symptoms of HCP. Microsurgical tumor excision without preoperative shunting was performed in 23 patients, 5 of whom required postoperative shunting in the first 2 months after tumor excision. Eighteen patients (78%) did not need shunts after tumor resection. With regard to tumor size, the postoperatively shunted group did not differ from the patients who had surgery but did not require shunt treatment (P < 0.50). The remaining 10 patients with preoperative HCP received shunts as the only treatment (3 patients), stereotactic radiosurgery (3 patients), or expectant management (4 patients). Two other patients without preoperative HCP developed postoperative HCP and required shunts. Postoperatively, we observed a significant (P < 0.001) increase in the incidence of pseudomeningocele and a nonsignificant (P < 0.1) increase in cerebrospinal fluid leaks (rhinorrhea and/or otorrhea) in patients without shunts as compared with postoperative patients without HCP. The patients were followed after any treatment for a mean of 3.2 years (range, 6 mo 10 yr). Follow-up in the patients who had surgery but did not require a shunt revealed a 61% decrease in clinical symptoms related to HCP and a 75% decrease in radiographic signs of HCP. CONCLUSION: In the presence of HCP, operative resection of CPA tumors can be performed without permanent cerebrospinal fluid shunting. Precautionary measures to decrease the incidence of postoperative complications related to cerebrospinal fluid leak in patients with preoperative HCP include meticulous obliteration of any exposed air cells, including those around the internal auditory canal, accurate restoration of the dural barrier, and temporary lowering of intracranial pressure with a ventricular or lumbar drain. Patients with persistent symptomatic HCP after tumor excision should be treated with a ventriculoperitoneal shunt. Delaying this decision until the postoperative period is safe and avoids unnecessary shunting in the majority of patients. PMID- 11383727 TI - High serum S100B levels for trauma patients without head injuries. AB - OBJECTIVE: Studies of patients with head trauma have demonstrated a correlation between a serum marker of brain tissue damage, namely S100B, and neuroradiological findings. It was recently demonstrated that the increases in serum S100B levels after heart surgery have extracerebral origins, probably surgically traumatized fat, muscle, and bone marrow. The current study examined multitrauma patients without head trauma, to determine whether soft-tissue and bone damage might confound the interpretation of elevated serum S100B concentrations for patients after head trauma. METHODS: A commercial assay was used to determine serum S100B concentrations for a normal population (n = 459) and multitrauma patients without head injury (n = 17). Concentrations of the two subtypes of S100B (S100A1B and S100BB) were determined using separate noncommercial assays. RESULTS: The mean serum S100B concentration for a normal healthy population was 0.032 microg/L (median, 0.010 microg/L; standard deviation, 0.040 microg/L). The upper 97.5% and 95% reference limits were 0.13 and 0.10 microg/L, respectively. No major age or sex differences were observed. Among trauma patients, serum S100B levels were highest after bone fractures (range, 2-10 microg/L) and thoracic contusions without fractures (range, 0.5-4 microg/L). Burns (range, 0.8-5 microg/L) and minor bruises also produced increased S100B levels. S100A1B and S100BB were detected in all samples. CONCLUSION: Trauma, even in the absence of head trauma, results in high serum concentrations of S100B. Interpretation of elevated S100B concentrations immediately after multitrauma may be difficult because of extracerebral contributions. S100B may have a negative predictive value to exclude brain tissue damage after trauma. Similarly, nonacute S100B measurements may be of greater prognostic value than acute measurements. PMID- 11383728 TI - Treatment of idiopathic trigeminal neuralgia: comparison of long-term outcome after radiofrequency rhizotomy and microvascular decompression. AB - OBJECTIVE: To evaluate the long-term outcome of patients after either percutaneous trigeminal rhizotomy or microvascular decompression (MVD) for idiopathic trigeminal neuralgia at a single institution. METHODS: From 1977 to 1997, 316 radiofrequency lesion procedures and 378 MVDs were performed. Questionnaires were sent to all patients who were alive in 1981, 1982, 1992, and 1998. For all other patients, interviews were conducted with their relatives and general practitioners. A retrospective comparative analysis was performed with Kaplan-Meier probability curves as of the latest follow-up date. In addition, 80 patients who underwent MVD were examined postoperatively with quantitative sensory measurements by use of von Frey hairs. RESULTS: Two hundred twenty-five patients who underwent MVD and 206 patients who underwent radiofrequency could be analyzed retrospectively in detail. Overall, there was a 50% risk for recurrence of pain 2 years after percutaneous radiofrequency rhizotomy. Conversely, 64% of patients who underwent MVD remained completely pain free 20 years postoperatively. Patients without sensory impairment after MVD were pain free significantly longer than patients who experienced postoperative hypesthesia or partial rhizotomy. CONCLUSION: Because it is curative and nondestructive, MVD is considered the treatment of choice for trigeminal neuralgia in otherwise healthy people. In our study, it proved to be a more effective and long-lasting procedure for patients with typical trigeminal neuralgia than radiofrequency rhizotomy. Patients without postoperative sensory deficit remained pain free significantly longer, which is a strong argument against the "trauma" hypothesis of this procedure. PMID- 11383729 TI - Treatment of refractory pain after brachial plexus avulsion with dorsal root entry zone lesions. AB - OBJECTIVE: Significant numbers of patients experience intractable pain after brachial plexus root avulsions. Medications and surgical procedures such as amputation of the limb are often not successful in pain treatment. METHODS: Forty seven patients with intractable pain after traumatic cervical root avulsions were treated with dorsal root entry zone coagulation between 1980 and 1998. The dorsal root entry zone coagulation procedure was performed 4 months to 12 years after the trauma, and patients were monitored for up to 18 years (average follow-up period, 14 yr). RESULTS: Immediately after surgery, 75% of patients experienced significant pain reduction; this value was reduced to 63% during long-term follow up monitoring. Nine patients experienced major complications, including subdural hematomas (n = 2) and motor weakness of the lower limb (n = 7). Improved coagulation electrodes with thermistors that could produce smaller and more accurate lesion sizes, which were introduced in 1989, significantly reduced the number of complications. CONCLUSION: Central deafferentation pain that persists and becomes intractable among patients with traumatic cervical root avulsions has been difficult to treat in the past. Long-term follow-up monitoring of patients who underwent the dorsal root entry zone coagulation procedure in the cervical cord indicated that long-lasting satisfactory relief is possible for the majority of individuals, with acceptable morbidity rates. PMID- 11383730 TI - Microsurgery for cerebral arteriovenous malformations: a dissection technique and its theoretical implications. AB - Surgical resection of cerebral arteriovenous malformations is often complicated by persistent bleeding from the nidus, which is difficult to manage. It has often been mentioned that the difficulty of hemostasis is caused by the fragile and pathological nature of vessels of the nidus. This may be one reason, but I hypothesize that the dissection procedure, which involves coagulation of the surface of the nidus, may cause obstruction of intranidal drainage and that this may cause increased intranidal pressure, especially at the late stage of resection. In operating on patients with cerebral arteriovenous malformations recently, I made an effort not to coagulate the surface of the nidus, and this technique has diminished hemorrhage problems effectively. I illustrate this technique herein with a resected specimen and plastic casts of the lesion. PMID- 11383731 TI - Angioplasty of intracranial occlusion resistant to thrombolysis in acute ischemic stroke. AB - OBJECTIVE: Thrombolysis has been demonstrated to improve revascularization and outcome in patients with acute ischemic stroke. Many centers now apply thrombolytic therapy locally via intra-arterial infusion. One therapeutic benefit is the ability to cross soft clots with a guidewire and to perform mechanical thrombolysis. In some instances, reopened arteries reocclude as a result of either thrombosis or vasospasm. We report the use of balloon angioplasty during thrombolysis for acute stroke. METHODS: From June 1995 through June 1999, 49 patients underwent intra-arterial therapy for acute stroke. In this group, nine patients (seven men and two women) were treated with balloon angioplasty after inadequate recanalization with thrombolytic infusion. The mean age of these patients was 67.9 years. Nine matched control patients who underwent thrombolysis alone without angioplasty were chosen for comparison. RESULTS: In the group of nine patients who had angioplasty, the mean National Institutes of Health Stroke Scale score at presentation was 21.8 +/- 5.4. Four patients had residual distal occlusion after angioplasty, and one patient had a hemorrhagic conversion. Of the five patients in which recanalization was successful, none had reocclusion of the balloon-dilated vessel. The mean score at 30 days for the five survivors was 12.6 +/- 14.9, for an improvement of 7.0 +/- 14.2. Among the nine control patients, the mean score at presentation was 20.3 +/- 5.2; the mean score at 30 days for the five survivors was 19.4 +/- 7.7, for an improvement of 4.2 +/- 7.8. CONCLUSION: In our experience, balloon angioplasty is a safe, effective adjuvant therapy in patients who are resistant to intra-arterial thrombolysis. The use of balloon angioplasty may prevent reocclusion in a stenotic artery and permit distal infusion of thrombolytic agents. PMID- 11383732 TI - Value of constructive interference in steady-state three-dimensional, Fourier transformation magnetic resonance imaging for the neuroendoscopic treatment of hydrocephalus and intracranial cysts. AB - OBJECTIVE: To assess the value of constructive interference in steady-state, three-dimensional, Fourier transformation (CISS) magnetic resonance imaging in the endoscopic management of hydrocephalus and intracranial cysts. METHODS: CISS imaging and T2-weighted imaging were performed for 14 consecutive patients before and after fenestration procedures, using a flexible endoscope, to treat loculated or multiloculated hydrocephalus (4 patients), aqueductal stenosis or obstruction (4 patients), arachnoid cysts (4 patients), a cyst of the velum interpositum (1 patient), or an ependymal cyst (1 patient). Fifteen fenestration procedures were performed, including one reoperation. RESULTS: Preoperative CISS imaging demonstrated intracystic intraventricular septa not observed with conventional T2 weighted imaging for 11 of 15 procedures and provided better brain tissue/cerebrospinal fluid contrast, allowing better understanding of the cause of hydrocephalus and the nature of the cysts. CISS imaging and T2-weighted imaging were equally useful for monitoring postoperative changes in the sizes of ventricles or cysts and the presence of flow voids after third ventriculostomies. However, only CISS imaging clearly demonstrated the site of fenestration for six of the nine patients who underwent fenestration procedures. CONCLUSION: CISS imaging provides excellent cerebrospinal fluid/brain tissue contrast, allowing detailed study of the anatomic features of the ventricular system and cystic lesions. CISS imaging is valuable for both preoperative decision-making and postoperative evaluation. PMID- 11383733 TI - Implantation of a reservoir for refractory chronic subdural hematoma. AB - OBJECTIVE: Recurrence of chronic subdural hematoma is not rare. Among patients who experience recurrence, severe background disease may adversely influence the prognosis of chronic subdural hematoma. We treated patients with these refractory hematomas with an Ommaya cerebrospinal fluid (CSF) reservoir and analyzed the effectiveness of the treatment. METHODS: Sixteen patients with refractory chronic subdural hematoma were studied. These patients had severe diseases that adversely influenced the clinical course of chronic subdural hematoma, including cerebral infarction, liver cirrhosis, thrombocytopenia, severe Parkinsonism, severe heart disease, psychiatric disease, and spinocerebellar degeneration. All patients were treated initially in the standard fashion: evacuation of the hematoma followed by irrigation and drainage of the hematoma cavity. In each patient, an Ommaya CSF reservoir was implanted after the hematoma recurred. Whenever the volume of the hematoma either decreased very slowly or increased, the reservoir was punctured. RESULTS: The hematoma size decreased to less than 3 mm a median of 60 days after introduction of the reservoir. Postoperatively, 13 patients returned to their condition before the onset of hematoma. One patient died of myocardial infarction, and two patients with Parkinson's disease could not maintain their previous functional level; both remained in a partially dependent state. Complications consisted of minor bleeding in two patients and occlusion of the reservoir in two other patients. CONCLUSION: By use of this method, reoperation was avoided and the patients were mobile early in the postoperative period. This method was suitable for refractory chronic subdural hematoma accompanied by severe disease that adversely influenced the clinical course. PMID- 11383734 TI - Fluoroscopic frameless stereotaxy for transsphenoidal surgery. AB - OBJECTIVE: To assess the value of frameless fluoroscopy-guided stereotactic transsphenoidal surgery using the FluoroNav Virtual Fluoroscopy System (Medtronic Sofamor Danek, Inc., Memphis, TN). METHODS: Twenty consecutive patients undergoing transsphenoidal surgery for sellar lesions were assigned to transsphenoidal surgery with or without computer-assisted fluoroscopic image guidance using the FluoroNav system. Prospective data regarding patient age, sex, lesion characteristics, operative time, and treatment cost were obtained. RESULTS: Although patients in the FluoroNav group were, on average, 17 years younger than the patients in the control group, more patients with recurrent adenomas were treated in the image guidance group. No other significant differences between the groups were found. FluoroNav provided accurate, continuous information regarding the anatomic midline trajectory to the sella turcica as well as anatomic structures (e.g., sella, sphenoid sinus) in the lateral view. No patient required reversion to intraoperative videofluoroscopy. No statistically significant differences were found with regard to preincision setup time, operative time, or cost. FluoroNav allowed procedures to be performed with significantly fewer x-rays being taken. CONCLUSION: Fluoroscopic computer assisted frameless stereotaxy furnishes accurate real-time information with regard to midline structures and operative trajectory. Although it is useful in first-time transseptal transsphenoidal surgery, its primary benefit is realized in recurrent surgery. PMID- 11383735 TI - Remote adenoviral gene delivery to the spinal cord: contralateral delivery and reinjection. AB - OBJECTIVE: This study characterizes the distribution of adenoviral genes in the spinal cord after viral vector injection into the sciatic nerve. It also evaluates the ability of repeated adenoviral sciatic nerve injections to prolong gene expression in the spinal cord. METHODS: Rat sciatic nerves were unilaterally coinjected with the retrograde tracer Fluoro-Gold (Fluorochrome, Inc., Denver, CO) and the adenoviral vector Ad5RSVntLacZ. The distribution of adenoviral gene expression in the spinal cord was compared with that of Fluoro-Gold. Next, levels of gene expression in the sciatic nerve and spinal cord were compared after single and repeated injections of Ad5RSVntLacZ. Finally, remote spinal cord gene expression in naive animals was compared with expression in animals that had been pretreated with subcutaneous Ad5RSVntLacZ inoculation. RESULTS: Viral gene expression was detected in all quadrants of the spinal cord gray matter, whereas Fluoro-Gold was detected only in the ipsilateral ventral horn (n = 5). This remote delivery was blocked by sciatic nerve transection (n = 10). Viral gene expression occurred in the sciatic nerve after both initial and repeated injections, whereas remote gene expression in the spinal cord was observed only after primary sciatic nerve injection (n = 24; P < 0.003). As with repeated sciatic nerve injections, subcutaneous inoculation with Ad5RSVntLacZ blocked subsequent remote spinal cord gene delivery (n = 8; P < 0.05). CONCLUSION: Remote viral gene delivery occurs in neurons without direct sciatic nerve projections but is dependent on intact peripheral nerves. Repeated injections fail to boost spinal cord gene expression, because of immune recognition of reinjected virus. PMID- 11383736 TI - Inhibition of experimental vasospasm by pretreatment with ultraviolet light irradiation in a rat femoral artery model. AB - OBJECTIVE: Chronic cerebral vasospasm is resistant to conventional treatments despite recent advances in treatment modalities. We studied the preventive effect of ultraviolet (UV) irradiation on development of vasospasm and its mechanism in a rat femoral artery model. METHODS: The rat femoral artery model for vasospasm was used in this investigation (n = 108). The femoral arteries were divided into four groups: empty and no irradiation (control), UV irradiation (UV group), blood placement (VS group), and blood placement after UV irradiation (VS + UV group). Luminal area was measured, and smooth muscle cell counts in the medial layer of the vessel wall were obtained. An immunohistochemical study was performed with cross sections of fixed femoral arteries at 12 hours and 1, 3, 5, 7, and 49 days after blood placement. The rings of femoral arteries on Day 7 were subjected to pharmacological study. RESULTS: Pretreatment with UV irradiation (VS + UV group) resulted not only in significant inhibition of chronic vasospasm but also in a significant decrease in smooth muscle cells compared with the VS group on Days 5 and 7. The UV-treated arteries (UV and VS + UV groups) exhibited a significant number of Bax- and Bcl-2-positive cells on Days 5 and 7, but few CPP-32 positive cells were observed at the same time points. In the pharmacological study, contractile response to KCI or phenylephrine was reduced significantly in the UV treated arteries. CONCLUSION: These results imply that UV irradiation prevents chronic vasospasm and suggest that UV-induced cell death plays an important role in the preventive effect without causing complications during the chronic period. PMID- 11383737 TI - Expression of endothelial nitric oxide synthase after exposure to perivascular blood. AB - OBJECTIVE: Although nitric oxide (NO) has been implicated in the development of vasospasm after subarachnoid hemorrhage, little is known regarding the time course of NO synthesis in vessel wall after exposure to perivascular blood. This study measures temporal characteristics of changes in vessel wall NO synthesis. METHODS: Rat femoral arteries exposed to perivascular blood for 3, 5, or 7 days were assayed for the endothelial isoform of NO synthase (eNOS) by Western blot testing. Additionally, rat femoral arteries exposed to perivascular blood for intervals from 3 to 14 days were analyzed by means of immunohistochemistry for eNOS. RESULTS: Semiquantitative densitometry of femoral artery Western blots demonstrated a biphasic pattern of eNOS expression after exposure to perivascular blood. Compared with control arteries, eNOS expression increased at 3 days (53 +/ 36%), normalized at 5 days (-6 +/- 7%), and decreased by 7 days (-39 +/- 15%). Immunohistochemistry confirmed the changes in expression of immunoreactive eNOS in femoral endothelium during the first week after chronic perivascular blood exposure and apparent reduced eNOS immunostaining, which persisted up to 14 days after application of blood. CONCLUSION: The expression of endothelial-derived NO in rat femoral artery exposed to perivascular whole blood does not directly correlate with changes in vessel caliber during this interval. The biphasic expression of eNOS observed in these experiments highlights the complexity of processes occurring in the vicinity of the vessel wall during vasospasm and may be related to several mechanisms that modulate vessel tone and response to injury. PMID- 11383738 TI - A scanning technique to measure regional cerebral blood flow and oxyhemoglobin level. AB - OBJECTIVE: The application of a laser scanning technique to measure regional cerebral blood flow (CBF) and tissue hemoglobin oxygenation (HbO2) using the rat closed cranial window preparation is described. METHODS: Twenty-nine male Wistar rats were used to consecutively measure local CBF by laser Doppler flowmetry and tissue HbO2 by a microspectrophotometric method at multiple corresponding cortical locations. The scanning technique used a computer-controlled micromanipulator. Data from three experimental models are presented: the whisker stimulation model, the ischemia-reperfusion model, and the sinus-vein thrombosis model. Sequential changes in local CBF and HbO2 data before, during, and after stimulation, ischemia, and sinus thrombosis were examined. Data from predefined locations within the same region were correlated with the topographical location and then arranged in a three-dimensional image. RESULTS: In the whisker stimulation model, we found a disproportionate increase in CBF (32 +/- 12%) as compared with that of HbO2 (9 +/- 4%) during stimulation. In the ischemia reperfusion model, the three-dimensional image showed heterogeneous low CBF (depending on the area) and homogeneous HbO2 at a reduced level during ischemia and postischemic hyperperfusion. However, the range of oxygenation was normal after reperfusion. In the sinus-vein thrombosis model, drainage of the unsaturated blood via the collateral pathways was noted. CONCLUSION: The laser scanning technique is useful for visualizing sequential changes in hemodynamic metabolic interactions of cortical brain tissue. This technique can reveal phenomena not detected by traditional monitoring procedures. PMID- 11383739 TI - African neurosurgery: current situation, priorities, and needs. AB - OBJECTIVE: The beginning of a new millennium causes us to reflect on the state of neurosurgery in the African countries and to affirm that we are eager to provide quality health care for our people. This study is important because its main purpose is to assess progress toward achievements and to identify potential problems, so that remedial action can be taken. METHODS: To attain these objectives, the following steps were performed: planning, surveying, analyzing feedback, and reporting. We first formed the World Health Organization African Subcommittee. In a meeting held in Dakar on May 25, 1998, we agreed on a strategy and devised a statistical form to adopt. After the drafts were approved, they were distributed to 51 African countries. The survey was performed by representatives who spent 6 months analyzing and collecting data. RESULTS: The results were alarming. After data analysis, we classified the countries into four main groups with respect to the number of neurosurgeons, the equipment potential, and the training capacity. One of the main observations is the need for organization and structured training. Another weakness is the lack of medicosurgical equipment. Poor funding and a lack of resources are responsible for this situation. CONCLUSION: Overcoming these problems will require that African neurosurgeons target a number of priorities, i.e., devising local training programs and obtaining the necessary equipment. We appeal to national and international institutions to focus on these two points to ensure long-term results, including greater involvement of African countries in local training and greater cooperation in terms of technical support and funding. PMID- 11383740 TI - International education: 20 years after implementation of a training program. AB - Medical care in developing countries may be improved by various means. Advances in the medical education and training of specialists play an important role in improving health care systems. The results of the efforts of a neurosurgeon from North America who helped establish neurosurgical training in Taiwan are reported after a follow-up period of 20 years. PMID- 11383741 TI - Functional neuroanatomy in the pre-Hippocratic era: observations from the Iliad of Homer. AB - OBJECTIVE: To describe observations of neurological significance made in the Iliad of Homer and to interpret these relative to pre-Hippocratic concepts of health and disease in Ancient Greece. METHODS: English translations of the Iliad were analyzed for references of neurological significance, and the Homeric Greek was subsequently reviewed for accuracy. Findings are discussed in the context of ancient Greek ideas regarding anatomy and physiology, early descriptions and conceptualizations of the nervous system, ancient Greek theories concerning illness and disease, and the practice of medicine in the pre-Hippocratic era. RESULTS: Descriptions of injuries sustained by soldiers fighting in the Trojan War represent some of the earliest case histories of neurotrauma. Passages in the Iliad describe immediate death after penetrating head trauma with injury to the brain or the brainstem, make reference to clinical signs of brain injury, and mention neurological signs and symptoms after damage to the spinal cord, brachial plexus, and peripheral nerves. CONCLUSION: The Iliad of Homer contains many descriptions of traumatic injury to the nervous system and provides us with 3,000 year-old references to some of the basic principles of functional neuroanatomy. PMID- 11383742 TI - Combined oligodendroglioma/pleomorphic xanthoastrocytoma: a probable collision tumor: case report. AB - OBJECTIVE AND IMPORTANCE: We report the first case of combined oligodendroglioma and pleomorphic xanthoastrocytoma. CLINICAL PRESENTATION: A female college student and competitive cross-country runner presented to the Mayo Clinic at age 18 years with complaints of progressive headache and visual disturbances. Neuroimaging revealed a heavily calcified left parieto-occipital mass with focal enhancement and remodeling of the overlying calvarium. INTERVENTION: A histological examination of the nearly gross total resection specimen disclosed a low-grade oligodendroglioma discretely abutting a superficially situated focus of pleomorphic xanthoastrocytoma. Ten months thereafter, a recurrence was suspected on the basis of the detection of a new focus of nodular enhancement. This lesion stabilized after radiation therapy, and the patient is well, without apparent residual disease 2.8 years after surgery. CONCLUSION: This unique neoplasm is presumed to represent a collision tumor, its biological behavior being similar to that of oligodendroglioma alone. However, the possibility of an unusual, mixed oligoastrocytoma with pleomorphic xanthoastrocytoma as the astrocytic component cannot be excluded entirely. Potentially, both components have the capacity for recurrence and anaplastic transformation. PMID- 11383743 TI - Meningeal solitary fibrous tumor as an unusual cause of expohthalmos: case report and review of the literature. AB - OBJECTIVE AND IMPORTANCE: We report an extremely rare case of a patient with meningeal solitary fibrous tumor with orbital involvement presenting as unilateral exophthalmos. This rare tumor should be considered in the differential diagnosis for aggressive dural-based lesions. CLINICAL PRESENTATION: A 54-year old man presented with a protruded eyeball on the right side and left hemiparesis. Computed tomography and magnetic resonance imaging demonstrated a huge mass, markedly enhanced in and around the right anterior clinoid process, which extended to the orbit and middle cranial fossa. Cerebral angiography revealed a richly vascular tumor fed by branches of both the right external and internal carotid arteries. A presumptive diagnosis of meningioma or hemangiopericytoma was considered. INTERVENTION: The tumor was exposed through an orbitozygomatic approach. At surgery, the tumor was grossly firm to hard and had destroyed the dura, orbital roof, anterior clinoid process, temporal bone, and muscle. Histological analysis revealed that the tumor was composed of spindle cell proliferation in a collagen-rich background, but it exhibited regional variations. The vascular network demonstrated irregular vascular lumina with a "hemangiopericytoma-like" pattern. Histological features included high cellularity and a high degree of pleomorphism. Immunohistochemical analysis of the tumor demonstrated diffuse positive staining for CD34 and vimentin. The tumor displayed no positive staining for cytokeratin, epithelial membrane antigen, glial fibrillary antigenic protein, S-100 protein, and factor XIII. CONCLUSION: Meningeal solitary fibrous tumor is considered a unique pathological entity. Wider use of immunohistochemical screening should enable analysis of the real incidence of these tumors; larger series and longer follow-up duration will allow conclusions to be drawn regarding treatment and prognosis. Differential diagnosis is discussed and the literature is reviewed. PMID- 11383744 TI - Giant cavernous sinus teratoma: a clinical example of a rare entity: case report. AB - OBJECTIVE AND IMPORTANCE: Teratomas represent 0.5% of all intracranial tumors. These benign tumors have tissue representative of the three germinal layers: ectoderm, mesoderm, and endoderm. Most teratomas are midline tumors located predominantly in the sellar and pineal regions. Cavernous sinus location is very rare; only two purely intracavernous teratomas have been reported. CLINICAL PRESENTATION: A 14-year-old boy presented with a history of progressive right eye proptosis and visual acuity impairment, headaches, and a neuralgia-like facial pain in the right V1 distribution. A head computed tomographic scan and magnetic resonance imaging disclosed a large tumor filling the right cavernous sinus and extending into the ipsilateral middle fossa. These scans also demonstrated mixed signals derived from different tissues conforming to the tumor (fat, cartilage, muscle strands, bone, and a primordial tooth). Heterogeneous enhancement was seen after infusion of contrast medium. Significant bone erosion and remodeling was evident in the middle fossa floor and right orbit, with secondary proptosis. A presumptive diagnosis of mature teratoma was made. INTERVENTION: With the use of a right frontotemporal interfascial approach, a combined extra- and intradural dissection of the tumor was performed. The lesion entirely occupied the cavernous sinus, laterally displacing the Gasserian ganglion and trigeminal branches (predominantly V1 and V2). The internal carotid artery and Cranial Nerve VI were medially displaced by the tumor mass. The lesion was composed of different tissues, including hair, fat, cartilage, muscle, nerve-like tissue, bone, and a primordial tooth. The tumor was removed completely, and the pathological report confirmed the diagnosis of a mature teratoma. There was no evidence of recurrence at the 8-month follow-up examination. CONCLUSION: Because of the lesion's location in the lateral wall of the cavernous sinus, a total removal was achieved with the use of standard microsurgical techniques. Knowledge of the microanatomy is essential in treating intracavernous pathology. We present the third reported case of a giant mature teratoma of the cavernous sinus. PMID- 11383745 TI - Exclusively epidural arteriovenous fistula in the cervical spine with spinal cord symptoms: case report. AB - OBJECTIVE AND IMPORTANCE: We describe the case of an epidural arteriovenous fistula (AVF) in the cervical spine draining only into the epidural and paravertebral plexus. An entirely epidural AVF having such drainage is extremely rare. CLINICAL PRESENTATION: A 24-year-old man presented with a 4-month history of gradually progressive sensory and motor disturbances of the upper and lower extremities. Magnetic resonance imaging and magnetic resonance angiography revealed a peridural vascular lesion within the canal compressing the spinal cord from C5 to T2. Diagnostic angiography revealed a perimedullary and/or dural high flow AVF, fed mainly by branches of ascending cervical and deep cervical arteries. The fistula drained into the epidural and paravertebral venous plexus without reflux into intradural venous systems. INTERVENTION: Multiple feeders of the AVF were embolized with a Liquid coil and n-butylcyanoacrylate via a two-step procedure. One week after embolization, the AVF was surgically removed. CONCLUSION: Interesting points of this case were the exclusively epidural location of the lesion, the exclusively epidural drainage of the AVF, and the etiology of the symptoms. Venous drainage of the fistula had no relation to any dural or intradural veins. Initially, spinal cord and nerve root compression by extradural veins with varicose dilation seemed to cause the radiculopathy and/or the myelopathy, and subsequent myelopathy caused by spinal venous hypertension was believed to be the main etiology in this case. PMID- 11383746 TI - Far lateral disc excision at L5-S1 complicated by iliolumbar artery incursion: case report. AB - OBJECTIVE AND IMPORTANCE: Paramedial approaches to far lateral discs at the L5-S1 space joint have advantages but may also bring the surgical space closer to large branch arteries of the internal iliac artery. I report incursion into an iliolumbar artery that required laparotomy to control hemorrhage. Surgeons performing extraforaminal disc explorations at L5-S1 need to evaluate preoperative magnetic resonance imaging scans for an enlarged iliolumbar artery near the disc space. CLINICAL PRESENTATION: The patient presented with a right L5 radiculopathy and a far lateral disc at L5-S1, as indicated by analysis of her magnetic resonance imaging scan. INTERVENTION: A midline incision and an extraforaminal exposure were performed. Arterial bleeding occurred when an anular disc fragment was removed with a pituitary rongeur under direct vision with microscopic magnification. Emergency laparotomy demonstrated hemorrhage from a branch of the internal iliac artery 2 cm from its origin. CONCLUSION: Iliolumbar artery variants may be at the margins of extraforaminal disc exposure at L5-S1. Preoperative magnetic resonance imaging scans should be evaluated for this vasculature structure. PMID- 11383747 TI - Resection of a giant intracranial dural arteriovenous fistula with the use of low flow deep hypothermic cardiopulmonary bypass after partial embolization: technical case report. AB - OBJECTIVE AND IMPORTANCE: To describe the surgical resection of a giant intracerebral arteriovenous fistula with involvement of dura mater and surrounding bone. Intraoperative bleeding was controlled by hypothermic circulatory arrest. CLINICAL PRESENTATION: This 46-year-old woman complained of persistent headache for 1 year; her diagnostic workup revealed the presence of an arteriovenous fistula in the dura mater of the left temporal region fed by the meningeal artery of the external and internal carotid arteries, with normal run off into Labbe's and Trolard's veins. Magnetic resonance imaging depicted a Chiari I malformation that was most likely a result of insufficient cerebral venous drainage. INTERVENTION: In preparation for surgery, staged embolization of feeders from the left meningeal artery and the left occipital artery was performed under controlled hypotension. This procedure failed to achieve a significant reduction in flow because of the immediate recruitment of internal branches of the internal carotid artery and dural branches of the right external carotid artery. Surgical treatment was undertaken without further embolization. Because of involvement of surrounding bone and the high risk of uncontrollable bleeding, the procedure was carried out with the patient under deep hypothermic cardiopulmonary bypass. Forty-five minutes of low flow (1.5 L/min) at 18 degrees C allowed total resection of the involved dura mater and surrounding bone. Postoperative recovery was marked by left brain edema that disappeared within 10 days. Findings on follow-up angiography were normal, and the patient was discharged with no neurological deficit. CONCLUSION: Low-flow deep hypothermic cardiopulmonary bypass can be used to control intraoperative bleeding for surgical excision of a giant intracerebral dural arteriovenous fistula. PMID- 11383748 TI - Improved cerebral perfusion and metabolism after stenting for basilar artery stenosis: technical case report. AB - OBJECTIVE: Recent advances in stent technology have allowed the introduction of more flexible stents that may be tracked more easily in the intracranial vessels. We present a patient with improved cerebral blood flow and metabolism as assessed by positron emission tomography after stent-assisted angioplasty for symptomatic basilar artery stenosis. CLINICAL PRESENTATION: A 62-year-old man, who had undergone left superficial temporal artery to middle cerebral artery bypass surgery for left internal carotid artery occlusion 10 years previously, presented with dizziness, blurred vision, and memory disturbance. Angiography revealed severe stenosis of the proximal basilar artery. Positron emission tomographic scans revealed decreased cerebral blood flow associated with increased oxygen extraction fraction in the entire brain, particularly in the posterior circulation and the left middle cerebral artery territory. Despite medical treatment, the patient experienced worsening visual disturbance and right-sided motor weakness. INTERVENTION: Ticlopidine and aspirin were used as antiplatelet agents. In addition, we used argatroban, which is a direct thrombin inhibitor, as an anticoagulant during the procedure. Predilation with a coronary artery balloon was performed, followed by placement of a GFX 3- by 8-mm stent (Arterial Vascular Engineering, Santa Rosa, CA), with excellent angiographic results. The patient made a good neurological recovery, and the postoperative positron emission tomographic scan demonstrated increases in both cerebral blood flow and cerebral metabolic rate of oxygen with a normalization of oxygen extraction fraction. CONCLUSION: Stent-assisted angioplasty can provide a favorable clinical course as well as improved cerebral perfusion and metabolism for a patient with basilar artery stenosis. Long-term follow-up data and additional clinical experience are required to assess the durability of this approach. PMID- 11383749 TI - Minocycline reduces traumatic brain injury-mediated caspase-1 activation, tissue damage, and neurological dysfunction. AB - OBJECTIVE: Caspase-1 plays an important functional role mediating neuronal cell death and dysfunction after experimental traumatic brain injury (TBI) in mice. Minocycline, a derivative of the antibiotic tetracycline, inhibits caspase-1 expression. This study investigates whether minocycline can ameliorate TBI mediated injury in mice. METHODS: Brains from mice subjected to traumatic brain injury underwent immunohistochemical analyses for caspase-1, caspase-3, and a neuronal specific marker (NeuN). Minocycline- and saline-treated mice subjected to traumatic brain injury were compared with respect to neurological function, lesion volume, and interleukin-1beta production. RESULTS: Immunohistochemical analysis revealed that activated caspase-1 and caspase-3 are present in neurons 24 hours after TBI. Intraperitoneal administration of minocycline 12 hours before or 30 minutes after TBI in mice resulted in improved neurological function when compared with mice given saline control, as assessed by Rotarod performance 1 to 4 days after TBI. The lesion volume, assessed 4 days after trauma, was significantly decreased in mice treated with minocycline before or after trauma when compared with saline-treated mice. Caspase-1 activity, quantified by measuring mature interleukin-1beta production by enzyme-linked immunosorbent assay, was considerably increased in mice that underwent TBI, and this increase was significantly diminished in minocycline-treated mice. CONCLUSION: We show for the first time that caspase-1 and caspase-3 activities localize specifically within neurons after experimental brain trauma. Further, these results indicate that minocycline is an effective pharmacological agent for reducing tissue injury and neurological deficits that result from experimental TBI, likely through a caspase-1-dependent mechanism. These results provide an experimental rationale for the evaluation of minocycline in human trauma patients. PMID- 11383750 TI - On individual preferences and aggregation in economic evaluation in healthcare. AB - For practical reasons, in order to carry out economic evaluations of collective decisions, total costs will generally be compared with total benefits; hence, individuals' willingness to pay (WTP) or quality-adjusted life-years (QALYs) have to be estimated at an aggregate level. So far, aggregation has usually been done by taking the individuals' mean WTP or the unweighted number of QALYs. Since the aggregation process is closely related to the way that income, health and/or utility of different individuals are compared and weighted, it also has significant equity implications. Thus. the explicit (or, more often, implicit) assumptions behind the aggregation process will largely affect how health and welfare are distributed is society. The aggregation problem in economic evaluation is certainly not trivial, but is seldom addressed in current practice. This paper shows the underlying assumptions of aggregate cost-benefit analysis (CBA) and cost-effectiveness analysis/cost-utility analysis (CEA/CUA), and it emphasises the particularly strong assumptions which have to be made when QALYs are interpreted as utilities in the welfare economics sense. Naturally, the appropriate method to choose depends on what is to be maximised: welfare or health. If decisions of resource allocation are to be based on economic welfare theory, then CBA should be preferred. However, if QALYs are interpreted as measures of health, rather than as utilities, then CEA/CUA would be appropriate. PMID- 11383753 TI - Augmenting reference pricing of pharmaceuticals in New Zealand with strategic cross-product agreements. AB - Reference pricing pharmaceuticals in New Zealand involves reimbursing drugs at the lowest price ruling in a given therapeutic subgroup, and has been argued to promote competition leading to equalised prices among similar drugs. Disappointment at the inability to contain public drug expenditures sufficiently has led to the augmentation of reference pricing with cross-product strategic agreements. These require firms seeking subsidisation of new drugs to significantly reduce their prices in unrelated markets, typically for relatively unpopular drugs. An examination of the markets for statins and angiotensin converting enzyme inhibitors in New Zealand shows that in neither case has price matching voluntarily accompanied these agreements. Although imperfect drug substitutability appears to be an important factor in explaining some of these results, particularly for statins, intrafirm cross-subsidisation induced by agreements and industry concern about international benchmarking of drug prices are proposed as major likely sources of influence. PMID- 11383751 TI - Pharmacoeconomics of hypertension management: the place of combination therapy. AB - Pharmacological treatment of hypertension has been shown to reduce the risk of stroke, coronary events, heart failure and progression of renal disease. However, rates of successful blood pressure control remain low among treated patients while antihypertensive medication represents a large and increasing proportion of healthcare expenditure in many countries. Several influential pharmacoeconomic analyses have confirmed the cost effectiveness of conventional antihypertensive treatments, usually involving monotherapy with diuretics or beta-blockers, compared with alternative strategies. Recent research has shown that a considerable proportion of the total cost of antihypertensive treatment in general practice is due to factors such as inadequate blood pressure control, poor compliance with therapy, discontinuation and switching between therapies. These factors operate to a much lesser extent in well-conducted clinical trials, and have not been fully incorporated into most economic studies. Some novel strategies, particularly low dose combinations of antihypertensive agents, may offer advantages in terms of efficacy, reduced adverse effects and improved compliance with treatment. There is therefore a need for comprehensive pharmacoeconomic analyses of novel strategies, taking these additional factors into account. Until such studies are available, the wider use of low dose combination therapy and other novel strategies should not be held back on the basis of earlier economic studies that have not included all relevant considerations. PMID- 11383754 TI - The cost of treatment of Alzheimer's disease in The Netherlands: a regression based simulation model. AB - OBJECTIVE: To examine the potential economic impact of treatment of Alzheimer's disease. DESIGN: Regression-based simulation estimation of the long term costs of Alzheimer's disease under a number of treatment scenarios. Data from an epidemiological study conducted in Rotterdam, The Netherlands, was used to simulate disease progression. Comparison of the costs and effectiveness experienced by the patients were used to measure the impact of treatment. PATIENTS AND INTERVENTION: 2 theoretical cohorts of patients with Alzheimer's disease, one of which receives standard treatment, while the other receives a treatment which slows cognitive decline as measured by the Mini-Mental State Examination (MMSE). MAIN OUTCOME MEASURES AND RESULTS: Under one of the scenarios examined, the baseline cost of Alzheimer's disease was 97,866 euro (EUR; 1996 values) per patient over 10 years' follow-up; the cost was almost EUR100,000 under all scenarios. Life expectancy following onset was about 4.5 years and MMSE decline was approximately 2 points per year for a typical prevalent (existing) patient and almost twice as much for incident (newly diagnosed) patients (1.82 vs 3.42 points per year, respectively). Slowing the rate of cognitive decline results in a slightly increased life expectancy, with more time being spent at home and less in a nursing home. Total costs (excluding those of therapy) will decrease, but savings will be modest and may well be less than the cost of therapy. Under the same scenario, total savings were EUR1,571 per patient which corresponds to an annual break-even cost of just EUR453. Decisions regarding the initiation or termination of therapy will affect both the number of patients treated and the costs and potential savings of treatment. CONCLUSIONS: The savings made in treating Alzheimer's disease will almost certainly be small in comparison with total costs and may well be offset by the cost of the treatment itself. Simulation models can be used to estimate the effect of therapy on the costs of care and can be useful tools in clinical decision-making and allocation of resources. These results show the need for further research into the costs and effects of treatment of Alzheimer's disease. PMID- 11383755 TI - Cost-benefit analysis of a Haemophilus influenzae type b meningitis prevention programme in The Philippines. AB - BACKGROUND: Haemophilus influenzae type b (Hib) meningitis is associated with high mortality and serious sequelae in children under 5 years of age. Vaccines which can prevent this infection are available. OBJECTIVE: To evaluate the costs and benefits of a 3-dose immunisation schedule in Manila, Philippines. PERSPECTIVE: Government and societal perspectives. DESIGN AND PARTICIPANTS: A cost-benefit analysis based on a birth cohort of 100,000 children. The state of health of the cohort with and without a Hib immunisation programme was modelled over a 5-year period. A survey of medical records of patients with Hib in Manila provided data on the extent and cost of sequelae following infection. INTERVENTION: A 3-dose Hib vaccination programme given at ages 2, 3 and 4 months. RESULTS: The model predicted that vaccinating children against Hib meningitis would prevent 553 cases per year in a birth cohort of 100,000, at a cost of 56,200 Philippine pesos (PHP) [$US1,605; 1998 exchange rate] per case (base case assumptions of 90% vaccine efficacy rate, 95 per 100,000 Hib incidence rate, 85% vaccination coverage). Results from the cost-benefit analyses indicated that the saving to the government would be around PHP39 million ($US1.11 million), and the saving to society would be PHP255 million ($US7.28 million). CONCLUSION: There would be a positive economic benefit for the Philippine government and for the Filipino society if a Hib vaccination programme was introduced in Manila. PMID- 11383752 TI - Symptom and health-related quality-of-life measures for use in selected gastrointestinal disease studies: a review and synthesis of the literature. AB - Patient-rated symptom and health-related quality-of-life (HR-QOL) outcomes are important end-points for clinical trials of medical treatments for gastrointestinal (GI) disorders. Based on this review, patient outcomes research is focused on gastroesophageal reflux disease and dyspepsia, with a growing interest in irritable bowel syndrome but little research in gastroparesis. State of-the-art for patient-rated symptom scales is rudimentary with an abundance of scales and little attention to systematic instrument development or comprehensive psychometric evaluation. Generally, disease-specific HR-QOL measures have been more systematically developed and evaluated psychometrically, but few have been incorporated into clinical trials. More comprehensive outcome assessments are needed to determine the effectiveness of new medical treatments for functional GI disorders. Future clinical trials of GI disorders should combine clinician assessments of outcomes and symptoms with patient-rated symptom and HR-QOL end points. PMID- 11383756 TI - Cost effectiveness of continuous terbinafine compared with intermittent itraconazole in the treatment of dermatophyte toenail onychomycosis: an analysis of based on results from the L.I.ON. study. Lamisil versus Itraconazole in Onychomycosis. AB - OBJECTIVE: To compare the costs and effectiveness of 2 oral antifungal treatment regimens in patients with dermatophyte toenail onychomycosis. DESIGN AND METHODS: A cost-effectiveness analysis using a model based on data from the Lamisil versus Itraconazole in Onychomycosis (L.I.ON.) study, a randomised controlled trial comparing continuous terbinafine with intermittent itraconazole. The trial included 4 treatment arms: terbinafine 250 mg/day for 12 or 16 weeks (T12, T16) and itraconazole 400 mg/day for 1 week in every 4 weeks for 12 or 16 weeks (I3, I4). Cost calculations for 6 countries (Finland, Germany, Iceland, Italy, The Netherlands, UK) included costs for medication, physician visits, laboratory tests, management of adverse events and management of relapse. Effectiveness was based on complete cure rates (mycological cure plus 100% toenail clearing). Costs per complete cure were determined and both average and incremental cost effectiveness ratios were calculated. PERSPECTIVE: Healthcare system. MAIN OUTCOME MEASURES AND RESULTS: In the L.I.ON. study, terbinafine was seen to be more effective than itraconazole (cure rates, 45.8 vs 23.4%). In most comparisons (5 of the 6 countries), the costs of T12 were statistically significantly lower than those of I3 [range: -37 to -173 euros (EUR); 1998 values; 1.172 US dollars = EUR1], indicating that T12 was the dominant strategy (i.e. less expensive and more effective). One exception (Finland) showed an incremental cost-effectiveness ratio of EUR524 per additional cure. In the other 5 countries, T16 and 14 were essentially equal in cost, but the greater effectiveness of T16 (cure rates, 55.1 vs 25.9%) resulted in a situation of extended dominance. CONCLUSION: From a healthcare system perspective, continuous terbinafine is less costly and more effective than intermittent itraconazole in the treatment of dermatophyte toenail onychomycosis. PMID- 11383757 TI - Cost-effectiveness analysis of n-3 polyunsaturated fatty acids (PUFA) after myocardial infarction: results from Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto (GISSI)-Prevenzione Trial. AB - OBJECTIVE: To estimate the cost effectiveness of treatment with n-3 polyunsaturated fatty acids (PUFA) for secondary prevention after myocardial infarction (MI). DESIGN AND SETTING: The cost-effectiveness analysis of n-3 PUFA treatment after MI was based on morbidity and mortality data and the use of resources obtained prospectively during the 3.5 year follow-up period of the Gruppo Italiano per lo Studio della Sopravvivenza nell'Infarto (GISSI) Prevenzione study. The cost-effectiveness analysis took into account the incremental number of life-years gained and the incremental costs for hospital admissions, diagnostic tests and drugs, applying a 5% discount rate. The value for money of n-3 PUFA treatment was assessed using the cost-effectiveness ratio and the number needed to treat (NNT) approach. PERSPECTIVE: Third-party payer. MAIN OUTCOME MEASURES AND RESULTS: The incremental cost-effectiveness ratio for n 3 PUFA in the basecase scenario was 24,603 euro (EUR, 1999 values) per life-year gained (95% confidence interval: 22,646 to 26,930). Sensitivity analysis included the analysis of extremes, producing estimates varying from EUR15,721 to EUR52,524 per life-year gained. 172 patients would need to be treated per year with n-3 PUFA, at an annual cost of EUR68,000, in order to save 1 patient. This is comparable with the NNT value, and associated annual cost for simvastatin, but less costly than that for pravastatin. CONCLUSIONS: The cost effectiveness of long term treatment with n-3 PUFA is comparable with other drugs recently introduced in the routine care of secondary prevention after MI. Since the clinical benefit provided by n-3 PUFA is additive, this therapy should be added to the established routine practice, with additive costs. PMID- 11383758 TI - Efavirenz: a pharmacoeconomic review of its use in HIV infection. AB - Efavirenz is a non-nucleoside reverse transcriptase inhibitor (NNRTI) used in the treatment of patients with HIV infection. Both US and British treatment guidelines for HIV infection recommend NNRTI- or protease inhibitor-based combinations [i.e. with nucleoside reverse transcriptase inhibitors (NRTIs)] as first-line treatmentoptions in the management of HIV disease. Results of a pivotal randomised study (DMP 266-006) comparing efavirenz- versus indinavir based triple combination therapy in patients with HIV infection (the majority of whom were antiretroviral therapy-naive) showed the efavirenz-based regimen was better tolerated and had greater success in achieving reductions in viral load below the limit of detection. These and other clinical data were incorporated into economic models in 2 analyses, one conducted in the US and the other in Canada. The US analysis examined long term clinical and economic outcomes predicted on the basis of response (viral load and CD4+ cell counts), tolerability and willingness to adhere to therapy. The efavirenz-based regimen was the dominant treatment strategy as it was predicted to improve survival and reduce direct medical costs in the US healthcare system. Compared with the indinavir-containing regimen, survival was increased by 11% (absolute difference) and cumulative costs were reduced by $US10,326 per patient (1998 discounted costs) at 5 years after starting treatment with efavirenz-based therapy. The Canadian analysis was conducted from the perspective of the Ontario healthcare system. This study did not consider differences in clinical efficacy between treatment groups, costs of study medication or outcomes beyond 1 year--all factors that would have favoured the efavirenz-based regimen. Of the 2 treatment options, the efavirenz-based regimen was associated with 7.4% lower average annual medical care costs, primarily because of greater costs associated with adverse clinical events with the indinavir-based regimen. In conclusion, current treatment guidelines for HIV infection recognise efavirenz-based combination regimens as a first-line treatment option. A pivotal comparative clinical trial (DMP 266-006) showed a significantly greater virological response to efavirenz- than indinavir-based triple combination therapy, and the efavirenz-based regimen was better tolerated. These clinical data are supported by pharmacoeconomic analyses conducted in the US and Canada, both of which showed lower medical care costs with the efavirenz-based regimen. The US analysis also predicted long term health benefits, such as improved survival, with efavirenz- versus indinavir based triple combination therapy. These results must be weighed against the inherent difficulties of predicting long term treatment failure rates from short term data, and the limited number of pharmacoeconomic analyses conducted with efavirenz to date. PMID- 11383759 TI - Will reversible (removable) refractive surgery reverse the way in which we do refractive surgery? PMID- 11383760 TI - Laser in situ keratomileusis and photorefractive keratectomy for residual refractive error after phakic intraocular lens implantation. AB - PURPOSE: To determine the visual and refractive outcome of photorefractive keratectomy (PRK) and laser in situ keratomileusis (LASIK) in eyes with prior posterior chamber phakic intraocular lens implantation for high myopia. METHODS: We studied a series of 37 consecutive eyes of 31 patients who underwent LASIK or PRK for residual refractive error following collamer posterior chamber intraocular lens (IOL) (Staar Surgical Implantable Contact Lens) implantation into a phakic eye. Twenty-eight eyes had LASIK and nine eyes had PRK. Mean follow up was 8.1 +/- 4.7 months after laser ablation (range, 3 to 18 mo). RESULTS: The preoperative mean spherical equivalent refraction prior to phakic posterior chamber IOL implantation was -17.74 +/- 4.89 D (range, -9.75 to -28.00 D). Following phakic IOL implantation and prior to LASIK or PRK, mean spherical equivalent refraction was -2.56 +/- 2.34 D (range, -0.25 to -8.75 D). One month following LASIK or PRK, mean spherical equivalent refraction was -0.24 +/- 0.52 D (range, -1.50 to +1.50 D), 3 months following LASIK or PRK, mean spherical equivalent refraction was -0.19 +/- 0.50 D (range, -1.50 to +1.00 D). The refraction was within +/-1.00 D of emmetropia in 36 eyes (97.2%) and within +/ 0.50 D in 31 eyes (83.7%). Three eyes developed anterior subcapsular opacities several weeks after laser ablation, one eye developed macular hemorrhage 4 weeks after laser ablation, and one eye had corticosteroid induced ocular hypertension. CONCLUSIONS: LASIK or PRK can be used to treat the residual refractive error following posterior chamber phakic IOL implantation. PMID- 11383761 TI - Geometric ray tracing analysis of visual acuity after laser in situ keratomileusis. AB - PURPOSE: Using a geometric ray tracing model, we explain the increase in visual acuity observed in myopic patients after laser in situ keratomileusis (LASIK). METHODS: This study included 37 eyes of 23 patients who underwent LASIK. All patients had myopia and a spectacle-corrected visual acuity of 0.95 or worse. Clinical tests included biometry, corneal topography, pachymetry, and refraction (with and without cycloplegia). Calculations were made by tracing rays through all the refractive surfaces of the eye based on a Le Grand-type theoretical model of the whole eye. RESULTS: Comparison of spectacle-corrected visual acuity of the eye before surgery, the size of the blur circle calculated by ray tracing, and the magnification for the ocular system facilitated a numerical criterion to assess visual acuity by geometric calculation. This criterion was applied to myopic eyes that underwent LASIK, and the maximum increase in spectacle-corrected visual acuity was predicted. An actual increase in visual acuity of approximately 40% of the predicted maximum was observed in patients. CONCLUSIONS: With geometric ray tracing, it was possible not only to obtain an estimate of the visual acuity before LASIK but also to assess the value of the maximum and probable increases in visual acuity after LASIK. PMID- 11383762 TI - Epithelial and stromal changes induced by intacs examined by three-dimensional very high-frequency digital ultrasound. AB - PURPOSE: To examine epithelial and stromal layers by three-dimensional very high frequency (VHF) digital ultrasound scanning before and after implantation of Intacs (intracorneal ring segments [ICRS]). METHODS: Three-dimensional scanning was performed in five eyes before and 3 months after Intacs insertion. Digital signal processing techniques provided high-resolution B-scan imaging and I-scan traces for high-precision (1-microm) three-dimensional pachymetry. Thickness maps of individual corneal layers were constructed of the epithelium, stroma, and full cornea before and after surgery. Difference maps for epithelium and stroma were produced to examine anatomical changes in the thickness profile induced in each layer and correlate these to refractive changes. RESULTS: B-scan examination revealed stromal and epithelial anatomy anterior and adjacent to the Intac. Ring depth could be measured topographically. There was stromal lamellar displacement by the ring segments that produced a concave anterior stromal groove within an annulus central to the ring. Epithelial filling of this concavity was shown in three dimensions in such a way as to produce orthogonally asymmetrical flattening of the corneal surface, thus potentially accounting for induced astigmatism. Mapping of the central stroma demonstrated thickening, potentially also accounting for astigmatic changes ascribable to orthogonal asymmetry. CONCLUSIONS: VHF digital ultrasound scanning provided imaging and three dimensional thickness mapping of corneal layers, enabling anatomical evaluation of the changes induced in the cornea by Intacs. PMID- 11383763 TI - Photorefractive keratectomy for myopia with the Meditec MEL 70G-Scan flying spot laser. AB - PURPOSE: To evaluate the efficacy, predictability, stability, and safety of the Meditec MEL 70G-Scan flying spot excimer laser. METHODS: One hundred thirty myopic eyes were treated with the Aesculap Meditec Mel 70G-Scan ArF flying spot excimer laser with photorefractive keratectomy (PRK). Patient groups: low myopia (Group 1) from -1.50 to -6.00 D (90 eyes), medium myopia (Group 2) from -6.10 to 9.00 D (31 eyes), high myopia (Group 3) from -9.10 to -14.00 D (9 eyes). RESULTS: At 12 months in the low myopia group, uncorrected visual acuity (UCVA) of 20/40 or better was achieved in 95.5% (86 eyes), 20/20 or better in 77.7% (70 eyes); 2.2% (two eyes) lost two or more lines of best spectacle-corrected visual acuity (BSCVA); 73.3% (66 eyes) were within +/-0.50 D of the target correction and 98.8% (89 eyes) were within +/-1.00 D. In the medium myopia group, UCVA of 20/40 or better was achieved in 74.2% (23 eyes), 20/20 or better in 25.8% (eight eyes); 3.2% (one eye) lost two lines, 61% (19 eyes) were within +/-0.50 D of desired correction and 83.8% (26 eyes) were within +/-1.00 D. In the high myopia group, UCVA of 20/40 or better was achieved in 22.2% (two eyes); 20/25 or better in 11.1% (one eye); none of the eyes achieved 20/20 UCVA; 22.2% (two eyes) lost two lines of BSCVA; 44.4% (four eyes) were within +/-0.50 D and 66.6% (six eyes) were within +/-1.00 D of the target correction. Refractive stability was achieved between 3 and 6 months. Increased intraocular pressure was detected overall in 7.69%. CONCLUSIONS: The results of predictability, safety, and efficacy in low and medium myopia with the Meditec MEL 70G-Scan flying spot excimer laser were good, but poorer predictability, regression, and a significant loss of BSCVA were observed in the high myopia group. PMID- 11383764 TI - Coefficient of friction of ocular surface lubricants for laser in situ keratomileusis. AB - PURPOSE: The coefficient of friction was empirically derived for a series of commercially available ophthalmic lubricants having potential use during laser in situ keratomileusis (LASIK). METHODS: Measurements were obtained using a modified tensile testing machine. Solutions tested included the artificial tear supplements Polytears-free, Bion Tears, Genteal, Refresh, Cellufresh, Celluvisc, Theratears, and Viscotears gel; and balanced salt solution (BSS). RESULTS: Viscotears carbomer gel had a significantly lower mean coefficient of friction compared to other ophthalmic lubricants tested (P=.002) and the dry plate (P=.005). Addition of 0.1 ml of BSS to 0.025 ml of Celluvisc significantly reduced the coefficent of friction of Celluvisc (P=.01). Polytears, Bion Tears, Genteal, Refresh, Celluvisc, Theratears, and BSS had coefficients of friction significantly higher than that of the dry plate. CONCLUSION: A range of values were obtained for the coefficient of the ophthalmic lubricants tested. Viscotears gel (0.1 ml), and Celluvisc (0.025 ml) + BSS (0.1 ml) had coefficients of friction significantly lower than that of the dry plate. PMID- 11383765 TI - Immunohistochemical study of subepithelial haze after phototherapeutic keratectomy. AB - PURPOSE: Subepithelial haze is a frequent complication and is often the cause of regression after photorefractive keratectomy (PRK). The lack of understanding of this undesirable complication following PRK is in part due to the limited availability of suitable tissues for pathological studies. METHODS: We examined the expression of various extracellular components in the cornea of a 46-year-old man who underwent phototherapeutic keratectomy (PTK) to remove a central corneal scar secondary to trauma. The patient subsequently underwent penetrating keratoplasty. A scar-free region containing an area of slight subepithelial haze adjacent to normal cornea was used for immunohistochemical staining with antibodies directed against cytoskeletal proteins, ie, vimentin, desmin and smooth muscle actin, and the extracellular components, laminin, heparan sulfate, keratan sulfate, and collagen types III, IV, V, and VII. RESULTS: Immunohistochemistry revealed that basal epithelial cells expressed components of basement membrane. The stromal fibroblasts within the haze tissue were labeled by anti-smooth muscle actin antibodies, a characteristic of myofibroblasts, which synthesized and secreted extracellular matrix components that contributed to the formation of the disorganized collagenous matrix and may account for subepithelial haze. CONCLUSIONS: The expression patterns for the cytoskeletal proteins and extracellular components indicated that the formation of subepithelial haze is a process of tissue remodeling, involving both corneal basal epithelial cells and keratocytes during wound repair. PMID- 11383766 TI - Mitomycin C reduces haze formation in rabbits after excimer laser photorefractive keratectomy. AB - PURPOSE: To investigate the effects of mitomycin C on haze after photorefractive keratectomy (PRK). METHODS: Twenty of 24 rabbits underwent bilateral 193-nm excimer laser PRK to correct -10.00 D of myopia; the remaining four rabbits were not operated (no PRK group). The right eyes of the 20 rabbits were treated with 0.02% mitomycin C during surgery (PRK+MMC group) and the left eyes did not receive 0.02% mitomycin C (PRK alone group). Clinical and histopathologic examinations were performed. RESULTS: The most severe haze in the PRK alone group after PRK reached grade 3; the PRK+MMC group did not exceed grade 1 haze. Statistically significant differences were found between the PRK+MMC and PRK alone groups from week 2 to week 26 after treatment (P<.01). Epithelial thickening appeared for 26 weeks in both PRK groups; no statistically significant differences were found between the two PRK groups (P>.05). A marked reduction of keratocytes in the anterior stroma of the PRK+MMC group was observed. At week 1, 2, and 4 after PRK, keratocytes of the PRK+MMC group were only 3.1+/-2.6, 6.8+/ 4.7, and 12.4+/-5.7 keratocytes x 10(4)/microm2, respectively, while those of the PRK alone group were 41.2+/-80, 42.3+/-7.8, and 40.0+/-3.3 keratocytes x 10(4)/microm2, respectively. There were statistically significant differences between the two groups (P<.001). CONCLUSION: A single intraoperative application of topical mitomycin C during PRK in rabbits reduced corneal haze by inhibiting the proliferation of keratocytes. PMID- 11383767 TI - Complications of laser in situ keratomileusis: etiology, prevention, and treatment. AB - PURPOSE: To review the etiology, prevention, and management of laser in situ keratomileusis (LASIK) complications. METHODS: Review of literature and the experience of the authors. RESULTS: Careful preoperative screening is critical to prevention of many potential complications of LASIK. Flap complications that occur during surgery are typically managed by replacement of the flap and repeating the surgery or applying special methods such as transepithelial photorefractive keratectomy weeks to months following the initial procedure. A common source of serious complications is the use of a microkeratome that functions after improper assembly. Timely treatment of postoperative complications such as diffuse lamellar keratitis, flap striae, and infection is critical to an optimal outcome. CONCLUSION: Most complications of LASIK can be treated effectively and have minimal effect on the final outcome after surgery, if appropriate methods are used for management. PMID- 11383768 TI - Muscular asthenopia and eccentric ablation after photorefractive keratectomy. AB - PURPOSE: To investigate the relationship between muscular asthenopia and eccentric ablation after photorefractive keratectomy (PRK). METHODS: Sixteen eyes of eight myopic patients whose muscular asthenopia was corrected by subjectively accepted spectacle prism after PRK, with visual acuity better than 0.8, were followed for 6 to 14 months. On the basis of preoperative and post-operative data and corneal topography, we calculated the total corrected corneal power using the Holladay formula and then measured the ablation eccentricity (h, millimeters) and its direction. According to the formula delta approximately equals Dh, the prism effective value (delta) caused by the eccentric ablation was computed and compared with the objectively accepted spectacle prism. RESULTS: The subjectively accepted spectacle prisms were similar to calculated values from the formula. Mean difference was 0.10 +/- 0.25delta. The direction of the subjectively accepted prism was in the direction of the ablation deviation. CONCLUSIONS: Eccentric ablation is an important cause of muscular visual asthenopia after PRK. The asthenopia may be corrected by spectacle prism. The spectacle prism value may be estimated by the formula delta approximately equals Dh. PMID- 11383769 TI - Interpreting the defocus equivalent index. PMID- 11383770 TI - Orbscan pachymetry after LASIK is not reliable. PMID- 11383771 TI - A case of failed LASIK. PMID- 11383772 TI - Tuning a Stradivarius. PMID- 11383773 TI - Evolving technology: recognition and opportunity. PMID- 11383774 TI - Thomas B. Ferguson lecture. Public policy and the practicing physician. PMID- 11383775 TI - High lactate predicts the failure of intraaortic balloon pumping after cardiac surgery. AB - BACKGROUND: Despite the use of intraaortic balloon pump (IABP) support in complex cardiac surgical patients, morbidity and mortality rates are high. More advanced mechanical cardiovascular support should be considered in those patients who are highly likely to die despite IABP support. We sought to identify early, readily available prognostic markers for patients receiving IABP support. METHODS: A retrospective analysis was performed on 39 patients requiring IABP support following cardiac surgery for more than 2 years. The accuracy and predictive ability of multiple potential markers of mortality were statistically assessed. RESULTS: Sixty-seven percent of the patients were successfully weaned from IABP support and 46% survived to hospital discharge. Serious complications occurred in 13% of patients. Serum lactate more than 10 mmol/L in the first 8 hours of IABP support predicted a 100% mortality. Base deficit more than 10 mmol/L, mean arterial pressure less than 60 mm Hg, urine output less than 30 mls/h for 2 hours, and dose of epinephrine or norepinephrine more than 10 microg/min were other highly predictive prognostic markers. CONCLUSIONS: Morbidity and mortality rates remain high despite IABP support following cardiac surgery. Mortality can be predicted by the presence of elevated serum lactate, elevated base deficit, hypotension, oliguria and large vasopressor doses, any of which should prompt appropriate consideration as to other mechanical cardiovascular support. PMID- 11383776 TI - An extracorporeal membrane oxygenation-based approach to cardiogenic shock in an older population. AB - BACKGROUND: We investigated the efficacy of an integrated system of advanced supportive care based on extracorporeal membrane oxygenation (ECMO) in older patients with an estimated mortality of more than 90% to establish whether its use is justifiable. METHODS: Treatment was provided by cardiac surgeons and critical care physicians and included the following key elements: (1) ECMO, (2) early application of continuous venovenous hemofiltration, (3) inhaled nitric oxide, (4) maintenance of perfusion pressure with norepinephrine, (5) maintenance of pulmonary blood flow by ventricular filling with intravenous colloids, (6) avoidance of early postoperative anticoagulation, (7) frequent use of transesophageal echocardiography, and (8) low tidal volume ventilation. Demographic features, intraoperative details, postoperative course, ECMO weaning rate, morbidity, survival to hospital discharge, and the quality of life of survivors were recorded. RESULTS: Seventeen consecutive patients (median age, 69 years) with refractory cardiogenic shock were studied. The median duration of ECMO was 86 hours (20 to 201 hours). Eleven patients (65%) were successfully weaned from ECMO. Seven patients (41%) survived to discharge. The major causes of morbidity were bleeding and leg ischemia. All patients who survived to discharge were alive and well at follow-up (median, 21 months) and reported a satisfactory quality of life. CONCLUSIONS: An ECMO-based approach can be used with acceptable results in the treatment of refractory cardiogenic shock, even in older patients. PMID- 11383777 TI - Risk factors for post-cardiopulmonary bypass vasoplegia in patients with preserved left ventricular function. AB - BACKGROUND: Although vasodilatory shock (VS) is one of the main complications of cardiopulmonary bypass (CPB), its pathophysiologic basis remains unclear. The aim of this study was to identify predisposing factors for the development of VS after CPB independent of ventricular function. METHODS: Thirty-six patients undergoing coronary artery bypass grafting who developed VS were compared with 72 control patients without post-CPB cardiogenic or vasoplegic shock, in a 2:1 case control study. Patients and controls underwent the same anesthetic protocol and were matched by age, sex, operation date, and left ventricle ejection fraction. RESULTS: Preoperative and intraoperative patient characteristics were not significantly different between the two groups. Preoperative use of angiotensin converting enzyme inhibitors and intravenous heparin were independent predictors for post-CPB VS by multivariate analysis (relative risk of 2.26 and 2.78, respectively). Intensive care unit stay and hospital stay were significantly longer in VS cases than controls, without any difference in early postoperative mortality. CONCLUSIONS: The only independent risk factors for postoperative VS identified were preoperative use of angiotensin-converting enzyme inhibitors and intravenous heparin. These risk factors were independent of age, gender, anesthetic protocol, and left ventricle ejection fraction. PMID- 11383779 TI - Total orthotopic heart transplantation for primary cardiac rhabdomyosarcoma: factors influencing long-term survival. AB - BACKGROUND: Primary cardiac sarcomas are uncommon and rare, with an unequal distribution in the population. A dismal prognosis is usually admitted that is related to a high propensity to develop distant metastasis with survival rarely exceeding 2 years. We report a case of a patient with a primary cardiac rhabdomyosarcoma characterized by an exceptional long-term survival after surgical treatment by a total orthotopic heart transplantation. From this limited experience, we reviewed factors that may influence survival to optimize therapeutic strategy. METHODS: A 33-year-old man was found to have a 10-cm primary cardiac rhabdomyosarcoma located in the right atrium and extending to the atrioventricular groove; therefore, resection was not possible. Since no metastases were detected, the patient was scheduled for urgent cardiac transplantation, which was performed after adjuvant radiotherapy. RESULTS: Postoperative outcome was uneventful and the patient is still alive, with regular follow-up, at 102 months. CONCLUSIONS: In a case of primary rhabdomyosarcoma, heart transplantation, despite immunosuppressive therapy, can provide long-term survival and can be considered for selected patients after rigorous analysis of predictors of survival. PMID- 11383778 TI - S100B as a predictor of size and outcome of stroke after cardiac surgery. AB - BACKGROUND: Stroke after cardiac surgery is a clinical problem with often fatal or disabling outcome. To assess severity and probable outcome in affected patients only from clinical and radiological examinations is difficult. The glial derived protein S100B has been suggested to be a marker of cerebral ischemia, and increased blood concentrations of S100B have been shown to correlate with size of lesion and prognosis after stroke. We studied the validity of S100B as a predictor of size of brain lesion and median term outcome in a consecutive group of patients suffering from stroke after cardiac surgery. METHODS: During a period of 17 months, 20 patients with clinical signs of postoperative stroke were investigated with S100B measurement, sampled at 5, 15 and 48 hours after surgery. All patients were examined with computed tomography or magnetic resonance imaging to confirm the diagnosis, and the size of cerebral infarction was estimated from the radiological examinations. The patients were followed up for survival 24 to 39 months after surgery. RESULTS: S100B concentration in blood 48 hours after surgery correlated with the size of infarcted brain tissue (r = 0.68, p < 0.001). Nine patients had S100B levels exceeding 0.5 microg/L and a 2-year mortality of 78%, whereas the 11 patients with S100B below 0.5 microg/L had a mortality of 18%. CONCLUSIONS: Increased S100B in patients with a stroke following cardiac surgery correlate with the size of infarcted brain tissue. High S100B levels 48 hours after surgery have a negative predictive value for median term survival. PMID- 11383780 TI - A multicenter, randomized, controlled trial of Celsior for flush and hypothermic storage of cardiac allografts. AB - BACKGROUND: A multicenter, randomized, controlled, open-label trial was conducted to evaluate the safety and efficacy of Celsior when used for flush and hypothermic storage of donor hearts before transplantation. METHODS: Heart transplant recipients were randomized to one of two treatment groups in which donor hearts were flushed and stored in either Celsior or conventional preservation solution(s) (control). Study subjects were followed for 30 days after transplantation. RESULTS: A total of 131 heart transplant recipients were enrolled (Celsior, n = 64; control, n = 67). The treatment groups were evenly distributed in donor and recipient base line characteristics. Graft loss rate was lower in the Celsior group on day 7 (3% versus 9%) and on day 30 (6% versus 13%), but the difference was not statistically significant based on 95% confidence interval analysis. No significant difference was measured between the Celsior and control groups in 7-day patient survival (97% versus 94%) and the proportion of patients with one or more adverse events (Celsior, 88%; control 87%) or serious adverse events (Celsior, 38%; control, 46%). Significantly fewer patients in the Celsior group developed at least one cardiac-related serious adverse event (13% versus 25%). CONCLUSIONS: Celsior was demonstrated to be as safe and effective as conventional solutions for flush and cold storage of cardiac allografts before transplantation. PMID- 11383781 TI - Surgical management of valvular disease in patients requiring left ventricular assist device support. AB - BACKGROUND: Success with long-term implantable left ventricular assist devices (LVAD) has led to increased use in patients previously thought to be unsuitable for mechanical circulatory assistance. Patients with preexisting or newly diagnosed valvular disease have been traditionally excluded from device placement. The purpose of this study was to review our experience with LVAD support in patients with valvular disease and to develop a management algorithm for these difficult patients. METHODS: We reviewed the clinical records of 199 consecutive patients who received the ThermoCardiosystems, Inc, HeartMate Pneumatic or Vented Electric LVAD. There were 18 patients (9%) who required surgical management of native or prosthetic valvular disease during LVAD implantation. RESULTS: Suture or patch closure of the aortic valve was performed in 6 patients, aortic valve plication and repair in 1 patient, mitral valve repair in 4 patients, and tricuspid valve annuloplasty in 5 patients. Two patients with mechanical mitral valve prostheses were treated with postoperative warfarin anticoagulation. Fifteen of the 18 patients with valvular pathology survived the immediate postoperative period (17% mortality). Eleven patients have either undergone transplantation or continue to be supported with an LVAD (61%). Operative mortality in LVAD patients without concomitant valve repair was 18% (n = 33) with a late mortality of 7% (n = 13). Seven of these late deaths occurred in patients who received a device as destination therapy. In the remaining 6 patients, the cause of death was sepsis (n = 2), multisystem organ failure (n = 2), driveline rupture (n = 1), and massive gastrointestinal bleed (n = 1). CONCLUSIONS: Preexisting native or prosthetic valve pathology does not increase the immediate perioperative risk of LVAD insertion; however, these patients continue to pose a challenge for postoperative management while awaiting transplantation. PMID- 11383782 TI - Favorable outcome after composite valve-graft replacement in patients older than 65 years. AB - BACKGROUND: Concomitant surgical replacement of the aortic valve and ascending aorta is an ideal treatment for aortic root aneurysms, but there may be hesitation in its use in older patients, despite their known increased risk of rupture. This study was conducted to examine our results in 84 patients older than 65 years undergoing elective aortic root resection with composite valve graft replacement. METHODS: Eighty-four patients older than 65 years were operated on between June 1987 and August 1998. Median age was 74 years (range, 66 to 89 years), and 57 patients were men. Seventeen patients were undergoing reoperation. Aortic insufficiency was present in 70 patients. Forty-seven patients received a conduit using a bioprosthesis, whereas in 37 a mechanical valved conduit (St. Jude) was used. The ascending aorta alone was replaced in 23 patients; 50 had hemi-arch replacement, and in 11 the entire aortic arch was replaced. RESULTS: Hospital mortality was 8.3% (7 of 84). Sixteen late deaths (19%) were noted during a median follow-up of 3.2 years (range, 0 to 10 years). Only one late death was aorta-related. The incidence of thrombotic or hemorrhagic complications was 2.1/100 patient-years, with equal frequency for both mechanical and bioprosthetic valves. CONCLUSIONS: We conclude that composite valve-graft replacement in elderly patients results in a low operative mortality, yields excellent long-term survival, and averts fatal aneurysm rupture in this high-risk population. PMID- 11383783 TI - Redo aortic root replacement: experience with 31 patients. AB - BACKGROUND: Aortic root re-replacement is being performed with increased frequency. Limited information is available regarding the surgical approaches and clinical outcomes of this reoperation. METHODS: Between May 1980 and May 1999, 31 patients (mean age, 45 +/- 15 years) underwent redo composite replacement of the aortic valve and ascending aorta. Indications for reoperation were prosthetic valve endocarditis in 12 patients (39%), failed biological valve in 17 (55%), and false aneurysm in 2 (6%). At reoperation, mechanical valves were implanted in 24 patients and biologic valves in 7. All patients with endocarditis had annular abscess and required reconstruction of the left ventricular outflow tract before implantation of a new valved conduit. Mechanical valves were used in 24 patients, aortic homograft in 4, and bioprosthetic valves in 3. The coronary button technique was used to reimplant the coronary arteries whenever possible. Extension of one or both coronary arteries with a short segment of saphenous vein or a synthetic graft was used in 16 patients (52%). The aortic arch was replaced in 7 patients (23%). RESULTS: There was one operative death (3%) because of rupture of an abdominal aortic aneurysm. The mean follow-up was 47 +/- 46 months and was 100% complete. There were five late deaths (16%), three of which were cardiac related. The actuarial survival was 71% +/- 12% at 5 years. Three patients experienced recurrent prosthetic valve endocarditis 4 months to 8 years after operation. The 8-year freedom from endocarditis for patients operated on for endocarditis was 82% +/- 11% compared with 100% for those operated on for other reasons (p = 0.1). At the last follow-up, 21 of 25 survivors (84%) were in New York Heart Association functional classes I or II, and 4 were in class III. CONCLUSIONS: Redo aortic root replacement can be performed with good early and late results. Patients operated on for prosthetic root endocarditis may have an increased risk of recurrent late endocarditis. PMID- 11383784 TI - Repair of anterior leaflet prolapse by papillary muscle repositioning: a new surgical option. AB - BACKGROUND: Although mitral valve repair is considered the gold standard for treating mitral regurgitation, anterior leaflet prolapse may still remain a challenging problem. This challenge is even greater for posterior commissural prolapse. We have used papillary muscle repositioning to treat anterior leaflet prolapse and suggest it as an alternative technique for all other methods previously described. METHODS: From 1989 to 1999 we performed 253 mitral valve repairs, among which 132 involved anterior leaflet prolapse. In this population there were two groups: group I (n = 92) treated with papillary muscle repositioning and group II (n = 40) treated with chordal shortening. There was no statistical difference between the two groups concerning age, functional class, and left ventricular function. Etiology was similar in both groups, a degenerative process being predominant. At echocardiography, regurgitation was graded 3.4/4 in both groups. There was no statistical difference concerning preoperative ejection fraction, end-systolic and end-diastolic left ventricular diameter. RESULTS: There were one in-hospital death in group I and two deaths in group II not related to mitral valve repair. Mean follow up is 36.4 +/- 29.2 months in group I and 70.5 +/- 9.5 months in group II. No patient was lost to follow-up. Mean regurgitation at follow-up was 0.75 +/- 0.67 in group I and 0.8 +/- 0.8 in group II (p = not significant). There was no statistical difference between the two groups concerning postoperative ejection fraction, end-systolic and end-diastolic left ventricular diameter. There was no late cardiac death in either group and there were no thromboembolic events. Actuarial survival rate is 98.9% and 96.3% in group I and 92.5% and 88.1% in group II at 3 and 8 years, respectively. CONCLUSIONS: Therefore, we conclude that papillary muscle repositioning is a safe technique that provides excellent results at mid-term follow-up and facilitates treatment of anterior leaflet prolapse. PMID- 11383785 TI - Midterm evaluation of the Tissuemed (Aspire) porcine bioprosthesis: 493 patients, 506 bioprostheses. AB - BACKGROUND: Valve durability has been a major concern with bioprostheses, and the Tissuemed (Aspire) porcine bioprosthesis was designed to provide a solution to structural valve failure. Because bioprostheses tend to fail by 8 years, the aim of our study was to determine its midterm durability and performance. METHODS: We reviewed 506 prostheses that were implanted in 493 patients (287 men; mean age 73 +/- 6 years) between 1991 and 1999. Preoperatively 316 (68%) patients were in New York Heart Association class III or IV. There were 417 (85%) aortic, 61 (12%) mitral, 13 (2.6%) aortic and mitral, and two (0.4%) tricuspid procedures. Concomitant procedures were performed in 163 (33%) patients. Follow-up was complete in 488 (98.9%) patients with a total cumulative follow-up of 1,402 patient-years. RESULTS: The 30-day mortality in this elderly population was 10% (95% confidence interval, 8 to 13), with no early valve-related deaths. Patients' survival at 8 years was 46% +/- 7%. This was influenced by the following factors: (1) the patient's age, being worse for those 70 years or older (p = 0.005); (2) those in New York Heart Association functional class III and IV (p = 0.004); (3) those in atrial fibrillation before the operation (p = 0.006); (4) those with poor left ventricular function (p = 0.009); and (5) those who had a previous cardiac operation (p = 0.003). Valve-related complications (expressed as percent per patient-year) were thromboembolism at 0.9%/patient-year; major hemorrhage at 1.4%/patient-year; bacterial endocarditis at 0.4%/patient-year; nonstructural dysfunction at 0.2%/patient-year, and reoperation at 0.2%/patient-year. At 8 years, freedom from thromboembolism was 93% +/- 7%, major hemorrhage, 90% +/- 4%, nonstructural dysfunction, 99% +/- 1%, structural valve failure, 100%, and reoperation, 99% +/- 1%. At follow-up, 98% of survivors were in New York Heart Association class I or II. CONCLUSIONS: Our study suggests that at 8 years, the Tissuemed (Aspire) porcine bioprosthesis is durable and has satisfactory performance with low complication rates. PMID- 11383786 TI - Clinical outcomes, angiographic patency, and resource utilization in 200 consecutive off-pump coronary bypass patients. AB - BACKGROUND: This retrospective study compared clinical outcomes and resource utilization in patients having off-pump coronary artery bypass grafting (OPCAB) versus conventional coronary artery bypass grafting (CABG). Angiographic patency was documented in the OPCAB group. METHODS: From April 1997 through November 1999, OPCAB was performed in 200 consecutive patients, and the results were compared with those in a contemporaneous matched control group of 1,000 patients undergoing CABG. Patients were matched according to age, sex, preexisting disease (renal failure, diabetes, pulmonary disease, stroke, hypertension, peripheral vascular disease, previous myocardial infarction, and primary or redo status. Follow-up in the OPCAB patients was 93% and averaged 13.4 months. RESULTS: Hospital death (1.0%), postoperative stroke (1.5%), myocardial infarction (1.0%), and re-entry for bleeding (1.5%) occurred infrequently in the OPCAB group. There were reductions in the rates of transfusion (33.0% versus 70.0%; p < 0.001) and deep sternal wound infection (0% versus 2.2%; p = 0.067) in the OPCAB group compared with the CABG group. Angiographic assessment of 421 grafted arteries was performed in 167 OPCAB patients (83.5%) prior to hospital discharge. All but five were patent (98.8%) (93.3% FitzGibbon A, 5.5% FitzGibbon B, 1.2% FitzGibbon O). All 163 internal mammary artery grafts were patent. Off-pump coronary artery bypass grafting reduced postoperative hospital stay from 5.7 +/- 5.3 days in the CABG group to 3.9 +/- 2.6 days (p < 0.001), with a decrease in hospital cost of 15.0% (p < 0.001). CONCLUSIONS: Off-pump coronary artery bypass grafting reduces hospital cost, postoperative length of stay, and morbidity compared with CABG on cardiopulmonary bypass. Off-pump coronary bypass grafting is safe, cost effective, and associated with excellent graft patency and clinical outcomes. PMID- 11383787 TI - Technical aspects and outcome of in situ right internal thoracic artery grafting to the major branches of the circumflex artery via the transverse sinus. AB - BACKGROUND: Little is known about the anatomic limitations of in situ right internal thoracic artery (RITA) grafting to the circumflex artery. METHODS: To evaluate the technical aspects and outcome of revascularization of the proximal and distal major branches of the circumflex artery (obtuse marginal [OM] branch and posterolateral [PL] branch), a total of 145 patients who possessed a graftable branch of the circumflex artery were enrolled into the prospective project. There were 73 patients who had the PL branch as a primary target and 72 patients with OM branches, which were allocated by a blinded observer who reviewed the preoperative angiography. RESULTS: Changes of primary target vessels were required in 9 patients (6.2%), yielding an overall success rate of RITA grafting of 93.8%. The success rates of RITA grafting to the OM branch and the PL branch were 95.8% (69/72; CI 88.3% to 99.1%) and 91.7% (67/73; CI 83.0% to 96.9%), respectively. The univariate analysis identified grafting under hypothermic ventricular fibrillation as predictors of inability to use in situ RITA grafting for revascularization of the circumflex artery. RITA grafting to the PL branch is not identified as a predictor. Postoperative angiography in 136 patients revealed only one occlusion (0.75%) of the RITA graft anastomosed to the marginal artery. There were no significant differences in patency rates between left and right ITA grafts. CONCLUSIONS: This prospective study showed that in situ RITA was, in most cases, able to reach most branches of the major circumflex artery and demonstrated an excellent patency rate. PMID- 11383788 TI - Atrial fibrillation: prevalence after minimally invasive direct and standard coronary artery bypass. AB - BACKGROUND: This study identified and compared the prevalence of new-onset atrial fibrillation (AFIB) following standard coronary artery bypass grafting (SCABG) with cardiopulmonary bypass (CPB) and minimally invasive direct vision coronary artery bypass grafting (MIDCAB) without CPB. A further comparison was made between AFIB prevalence in SCABG and MIDCAB subjects with two or fewer bypasses. METHODS: This is a retrospective, comparative survey. Patients with new-onset AFIB who underwent SCABG or MIDCAB alone were identified electronically using a triangulated method (International Classification of Diseases, 9th revision, Clinical Modification [ICD-9 CM] code; clinical database word search; and pharmacy database drug search). RESULTS: The total sample (n = 814; 94 MIDCAB, 720 SCABG) exhibited a trend toward lower AFIB prevalence in MIDCAB (23.4%) versus SCABG (33.1%) subjects (p = 0.059). AFIB prevalence in the SCABG subset with two or less vessel bypasses (n = 98; n = 18 single vessel, n = 80 double vessels) and MIDCAB subjects (n = 94; n = 90 single vessels, n = 4 double vessels) was almost identical (SCABG subset 24.5% versus MIDCAB 23.4%, p = 0.860). Slightly more than half (56.9%) of new-onset AFIB subjects were identified by ICD-9 CM codes, with the remainder by word search (37.7%) or procainamide query (5.4%). CONCLUSIONS: In this sample, the number of vessels bypassed seemed to have a greater influence on AFIB prevalence than the application of CPB or the surgical approach. Retrospective identification of AFIB cases by ICD-9 CM code grossly underestimated AFIB prevalence. PMID- 11383789 TI - Hemodynamics and gas exchange during carbon dioxide insufflation for totally endoscopic coronary artery bypass grafting. AB - BACKGROUND: In addition to single-lung ventilation (SLV), positive-pressure CO2 insufflation is mandatory for totally endoscopic coronary artery bypass grafting. Studies on the effects of unilateral CO2 insufflation on hemodynamics produced controversial results, and bilateral insufflation has not been studied to our knowledge. The present study sought to investigate hemodynamics and gas exchange during unilateral and bilateral CO2 insufflation in patients who underwent totally endoscopic coronary artery bypass grafting. METHODS: Eleven hemodynamic and gas exchange variables were monitored during 22 totally endoscopic coronary artery bypass grafting procedures with unilateral (n = 17) or bilateral (n = 5) CO2 insufflation at a pressure of 10 to 12 mm Hg. Data were obtained at baseline with double-lung ventilation, after institution of SLV, during insufflation, after cardiopulmonary bypass during SLV, and after return to double-lung ventilation. RESULTS: Arterial oxygen tension decreased significantly during SLV, whereas the peak inspiratory pressure increased. In addition, central venous pressure and heart rate increased significantly during insufflation, but mean arterial pressure remained unchanged. Although the end-tidal CO2 pressure did not change, arterial carbon dioxide tension increased progressively to a maximum of 44.6 +/- 5.9 mm Hg during unilateral insufflation, and 55.7 +/- 14.6 mm Hg during bilateral insufflation (p < 0.05 versus baseline and between groups). Mixed venous oxygen saturation declined during SLV regardless of CO2 insufflation and recovered to baseline once double-lung ventilation was restarted. Left and right ventricular ejection fractions remained unaltered. No patient required inotropic or vasopressor support. CONCLUSIONS: Carbon dioxide insufflation for totally endoscopic coronary artery bypass grafting with SLV had no adverse effects on hemodynamics. In contrast to a moderate increase of arterial carbon dioxide tension during unilateral insufflation, markedly elevated arterial carbon dioxide tension levels remain a cause of concern during bilateral insufflation. PMID- 11383791 TI - Is tranexamic acid safe in patients undergoing coronary endarterectomy? AB - BACKGROUND: Patients undergoing coronary endarterectomy during coronary artery bypass grafting (CABG) are at increased risk of perioperative myocardial infarction due to coronary intimal disruption. Data assessing the safety of the antifibrinolytic drug tranexamic acid (TA) in patients undergoing this procedure are lacking. METHODS: From September 1997 to December 1999, 221 patients underwent nonemergency primary CABG with endarterectomy of the right coronary artery alone in 149, the left anterior descending in 35, or both right and left anterior descending in 27. TA was administered intraoperatively to 87 patients (TA group: average total dose 62 +/- 4.4 mg/kg; range 20 to 109 mg/kg), and was not administered to 134 patients (No TA group). RESULTS: The patient characteristics of the 2 groups were similar. In-hospital mortality consisted of 2 patients in the TA group and 4 patients in the No TA group. Perioperative myocardial infarction rates were 2% and 5% in the TA and No TA groups, respectively (p = 0.49). The relative risk for any type of perioperative cardiac ischemic event in the TA group versus the No TA group was 0.77 (95% CI; 0.4, 1.2). Patients in the TA group had a significant reduction in postoperative chest tube drainage (685 versus 894 mL in the TA versus No TA groups, respectively) and in the use of fresh-frozen plasma (p = 0.03). CONCLUSIONS: These results suggest that the clinical effectiveness of tranexamic acid in reducing postoperative blood loss in patients undergoing coronary endarterectomy is not associated with a higher incidence of myocardial ischemia-related complications. PMID- 11383790 TI - Minimally invasive saphenous vein harvesting: effects on endothelial and smooth muscle function. AB - BACKGROUND: The long saphenous vein remains the commonest conduit used in coronary artery bypass grafting procedures. Surgical trauma during vein harvesting can cause endothelial and smooth muscle injury that has important implications for vein graft longevity. Minimally invasive vein harvesting is advocated to reduce wound morbidity. However, the functional consequences of increased handling and traction, with potentially detrimental effects, remain unknown. METHODS: Forty patients were prospectively randomized into either a minimally invasive (minimal) or traditional (open) saphenous vein harvest group. Smooth muscle contractile function was assessed by responses to potassium chloride and phenylephrine. Endothelial cell function was assessed by responses to serial escalations in concentration of acetylcholine, bradykinin, calcium ionophore, sodium nitroprusside, and N-nitro-L-arginine using isometric tension studies. RESULTS: Harvest times were similar for both groups. The total incision length in the minimal group was significantly shorter than in the open group. There were no differences in smooth muscle contractions to either receptor independent or receptor-mediated agonists between the two groups. Similarly, vasorelaxation in response to both endothelium-dependent and endothelium independent agonists were similar in both groups. CONCLUSIONS: Minimally invasive saphenous vein harvesting is associated with similar medial smooth muscle and endothelial function as open harvesting. These findings suggest that minimally invasive harvesting techniques can be used without major detrimental effects on vascular integrity. PMID- 11383792 TI - Increase in serum S100A1-B and S100BB during cardiac surgery arises from extracerebral sources. AB - BACKGROUND: Elevated levels of serum S100B after coronary artery bypass grafting may arise from extracerebral contamination. Serum S100B content was analyzed in several tissues, and the two dimers S100A1-B and S100BB were analyzed separately in blood. METHODS: Serum, shed blood, marrow, fat, and muscle were studied in patients undergoing coronary artery bypass grafting with cardiopulmonary bypass using suction either to the cardiotomy reservoir (group 1, n = 10) or to a cell saving device (group 2, n = 10), or operated on off-pump (group 3, n = 10). RESULTS: Serum S100B was sixfold higher in group 1 than in groups 2 and 3, which were identical. The same ratio between S100A1-B and S100BB was found in all groups. When compared with serum, S100B was 10(2) to 10(4) times higher in marrow, fat, muscle tissue, and shed blood. CONCLUSIONS: Separate analysis of S100A1-B and S100BB did not distinguish between S100B of cerebral and extracerebral origin. The concept that S100B only originates in astroglial and Schwann cells is wrong. Fat, muscle, and marrow in mediastinal blood contain high levels of S100B. Cardiopulmonary bypass caused no increase in S100B. PMID- 11383793 TI - Cardiopulmonary bypass induces the synthesis and release of matrix metalloproteinases. AB - BACKGROUND: A number of cellular and molecular events can be induced after cardiac procedures requiring cardiopulmonary bypass (CPB). The matrix metalloproteinases (MMPs) are a recently discovered family of enzymes that degrade the extracellular matrix, but expression during and after CPB is unknown. METHODS: Systemic plasma MMP levels were measured in patients (n = 28, 63 +/- 1 years) undergoing elective coronary revascularization requiring CPB at baseline, termination of CPB, and 30 minutes, 6 and 24 hours after CPB. Representative classes of MMP species known to degrade matrix and basement membrane components were selected for study. Specifically, the interstitial collagenases MMP-8 and MMP-13, and the gelatinases MMP-2 and MMP-9 were determined by internally validated enzyme-linked immunosorbent assay. RESULTS: The MMP-8 levels increased by fourfold at separation from CPB, and returned to within normal values within 30 minutes after CPB. The proenzyme forms of MMP-13 and MMP-9 increased by more than twofold at cross-clamp release and returned within normal limits within 6 hours after CPB. The proform of MMP-2 increased from baseline values at 6 and 24 hours postoperatively; likely indicative of de novo synthesis. CONCLUSIONS: A specific portfolio of MMPs are released and synthesized during and after CPB. Because MMPs can degrade extracellular proteins essential for maintaining normal cellular architecture and function, enhanced MMP release and activation may contribute to alterations in tissue homeostasis in the early postoperative period. PMID- 11383794 TI - Prophylactic use of pentoxifylline on inflammation in elderly cardiac surgery patients. AB - BACKGROUND: Inflammation plays a pivotal role in the pathogenesis of organ injury after cardiopulmonary bypass (CPB). Elderly patients appear to be especially prone to develop general inflammation. Use of pentoxifylline (PTX) before surgery may be a promising approach to minimize the negative effects of CPB in these patients. METHODS: In a prospective, randomized study, patients more than 80 years old undergoing aortocoronary artery bypass grafting received either PTX (n = 15) after induction of anesthesia (initial bolus of 300 mg followed by a continuous infusion of 1.5 mg.kg(-1).h(-1) during the next 2 days) or saline as placebo (control group; n = 15). Polymorphonuclear neutrophil (PMN) elastase, C reactive protein (CRP), and interleukins (IL-6, IL-8, IL-10) were measured from arterial blood samples before surgery (T0), at the end of surgery (T1), 5 hours after surgery (T2), and at the morning of the first (T3) and second (T4) postoperative day. RESULTS: Postoperatively, PTX-treated patients less often needed catecholamines and were extubated earlier than the control patients (p < 0.05). On the intensive care unit, cardiac index inceased more in the PTX-treated (from 1.95 +/- 0.3 to 3.26 +/- 0.4 L.min(-1).m(-2)) than in the control patients (from 1.89 +/- 0.2 to 2.78 +/- 0.3 L.min(-1).m(-2)). Increase in CRP and PMN elastase was significantly higher in the untreated control than in the PTX patients. After CPB, IL-6, IL-8, and IL-10 increased in both groups showing a significantly higher increase in the untreated control patients (IL-8 control: from 11.3 +/- 2.6 to 154.4 +/- 57 pg/mL [T1]); IL-8 PTX: from 10.9 +/- 2.7 to 71.8 +/- 23 pg/mL [T1]). CONCLUSIONS: In elderly cardiac surgery patients, use of PTX before surgery and continued after CPB resulted in less inflammatory response than in an untreated control group. The value of attenuating the inflammatory process by PTX on outcome in this patient population needs to be evaluated in further controlled studies. PMID- 11383795 TI - Reverse subclavian flap repair of hypoplastic transverse aorta in infancy. AB - BACKGROUND: Management of hypoplastic aortic arch associated with coarctation in infancy can be challenging. Reverse subclavian flap aortoplasty plus coarctation resection offers simplicity without needing foreign material or cardiopulmonary bypass. METHODS: Since 1988, 46 of 162 infants less than 3 months undergoing coarctation repair had hypoplastic arch enlargement with reverse subclavian flap aortoplasty. Median age was 11 days; mean weight was 3.2 kg. Thirty-seven patients (80%) had associated cardiac defects including single or multiple ventricular septal defects (14 infants), transposition of the great arteries (7), aortic or mitral stenosis (5), and complete atrioventricular septal defect (5 infants). Twenty-eight patients had pulmonary artery banding; 2 had an arterial switch operation through a separate median sternotomy. RESULTS: There were two hospital deaths: one 4 months postoperatively in a patient requiring a Norwood procedure the next day for underestimated left ventricular hypoplasia; the other of sepsis more than 1 month postoperatively. On follow-up from 1 to 129 months (mean, 38 months), there were five recurrent obstructions: three at the coarctation site treated with balloon dilatation and two at the arch site. Twenty six children had their heart defects corrected with 29 subsequent operations including an arterial switch operation for transposition of the great arteries/ ventricular septal defect (3 infants), relief of aortic or mitral stenosis +/- ventricular septal defect closure (5), multiple ventricular septal defect closure (3), a bidirectional Glenn (2), complete atrioventricular septal defect (2), and anomalous left coronary with ventricular septal defect repair (1 infant). Four children await debanding and ventricular septal defect closure or Glenn anastomosis. There have been two late deaths (overall survival, 91%). CONCLUSIONS: Reverse subclavian flap aortoplasty is excellent for relief of arch hypoplasia and coarctation in infants with low recurrence rates and acceptable operative and intermediate survival. PMID- 11383796 TI - Saphenous vein homograft: a superior conduit for the systemic arterial shunt in the Norwood operation. AB - BACKGROUND: Excessive pulmonary blood flow increases ventricular volume work in the face of inadequate systemic cardiac output, low diastolic blood pressure, and inadequate coronary perfusion. Using the smallest available 3-mm polytetrafluoroethylene shunts have been successful, although catastrophic shunt thrombosis has occasionally been observed. To avoid thrombosis with a smaller conduit, saphenous vein homografts (SVG) were used to construct the modified Blalock-Taussig (BT) shunts. METHODS: From January 1998 to April 1999, 25 patients weighing 3.1 kg (3.0 kg or less, n = 9), at a mean age of 8.9 days, underwent stage I Norwood using an SVG BT shunt. Common heart defects were aortic atresia (n = 8), mitral atresia and double-outlet right ventricle (n = 5), and unbalanced AVC (n = 5). Mean BT shunt size was 3.2 mm, with 12 patients having shunts that were 3 mm or smaller. RESULTS: Thirty-day hospital mortality was 8% (2 of 25). No shunt thrombosis was seen, despite banding the BT shunt in 3 patients. One patient had BT revision because of an anatomic issue not directly related to the shunt material. CONCLUSIONS: Excellent results may be achieved using SVG BT shunts in the Norwood operation. This conduit seems less likely to thrombose, both acutely and chronically, allowing the use of appropriately smaller-sized shunts in small neonates. PMID- 11383797 TI - Total cavopulmonary connections in children with a previous Norwood procedure. AB - BACKGROUND: Outcomes of the Fontan operation in children initially palliated with the modified Norwood procedure are incompletely defined. METHODS: From August 1993 to January 2000, 45 patients (mean age 2.6 +/- 1.1 years, weight 12.7 +/- 2.8 kg) who were palliated with staged Norwood procedures (hypoplastic left heart syndrome, n = 32; nonhypoplastic left heart syndrome, n = 13) underwent a modified Fontan operation. Preoperative features included moderate/severe atrioventricular valve regurgitation (n = 5, 11%), reduced ventricular function on echocardiography in 11 patients, McGoon index 1.56 +/- 0.38, and pulmonary artery distortion in 18 patients (40%). RESULTS: A lateral tunnel (n = 16) or an extracardiac conduit (n = 29) connection with fenestration in 38 patients (84%) was used. Concomitant procedures included pulmonary artery reconstruction (n = 24, 53%), atrioventricular valve repair (n = 4, 9%) or replacement (n = 1). Before Fontan, 12 patients (27%) had an intervention to address neoaortic obstruction, and 7 patients required balloon dilation/stenting of the left (n = 5) or right pulmonary artery (n = 5). Intraoperatively, left (n = 5) or right pulmonary artery (n = 1) stenting was performed in 5 patients (11%). On follow up, 8 patients required additional interventional procedures to address left pulmonary artery narrowing (n = 5), or venous (n = 5) or arteriopulmonary collaterals (n = 1). Perioperative mortality was 4.4% (n = 2). There were 2 late deaths at a mean follow-up of 39 +/- 20 months. CONCLUSIONS: In relatively high risk patients, midterm results of the Fontan operation for children initially palliated with the Norwood procedure were good. Combined interventional-surgical treatment algorithms can lead to improved outcomes. PMID- 11383798 TI - Cardiac arrhythmia in patients undergoing surgical repair of Ebstein's anomaly. AB - BACKGROUND: Arrhythmias remain an unsolved problem in Ebstein's anomaly. The aim of this study was to investigate the evolution of arrhythmias after surgical repair. METHODS: Forty-five patients with Ebstein's anomaly and arrhythmias were studied. Mean age was 33 +/- 15 years. Twenty-four patients (53%) had paroxysmal supraventricular tachycardia, 12 (27%) had atrial fibrillation or flutter, 8 (18%) had ventricular preexcitation (Wolff-Parkinson-White syndrome), and 1 (2%) had a nonsustained ventricular tachycardia. Surgical technique included detachment of the tricuspid anterior leaflet and suture on the atrioventricular annulus associated with right ventricular longitudinal plication. RESULTS: There were four hospital deaths (9%). A pacemaker was implanted early after operation in 5 patients (11%). During a mean follow-up of 57 +/- 50 months (range, 4 to 226 months), there were six additional deaths, three of which were sudden. Two patients were lost to follow-up. Of the 33 surviving patients, 8 (24%) continued to have symptomatic arrhythmias, and 15 (45%) were in permanent sinus rhythm. Of the 24 patients with preoperative paroxysmal supraventricular tachycardia and the 12 with atrial fibrillation or flutter preoperatively, 9 and 2 of the survivors, respectively, have had no further episodes of arrhythmia. The incidence of arrhythmia with or without symptoms was reduced to 39% (13/33) of the surviving patients. CONCLUSIONS: Arrhythmia is not totally abolished after operation. However, patients with Ebstein's anomaly and arrhythmia show substantial improvement after conservative surgical intervention. PMID- 11383799 TI - A policy of elective delayed sternal closure does not improve the outcome after arterial switch. AB - BACKGROUND: Delayed sternal closure is regularly used in the immediate management of hemodynamic instability after neonatal cardiac procedures. The aim of this study was to assess whether the routine, elective use of delayed sternal closure would reduce morbidity in neonates undergoing arterial switch for transposition of the great arteries. METHODS: A retrospective statistical analysis was performed on 52 neonates operated on from 1991 to 1998. Until 1994, chest closure was routinely attempted in all patients after arterial switch; the policy was then changed to delayed sternal closure in all cases in the latter half of the study period. RESULTS: Delayed sternal closure did not significantly alter the mean duration of ventilation (2.7 +/- 2.37 versus 2.7 +/- 1.3 days) nor intensive care stay (4.1 +/- 2.8 versus 5.7 +/- 10.0 days; p = 0.46). There was no increase in the incidence of wound sepsis (7.7% versus 3.8%; p = 0.55), and mortality was unchanged (7.7% in both groups). There was an increase in the incidence of urgent reexploration (7.7% versus 19.2%; p = 0.22), which did not reach significance. CONCLUSIONS: This study does not support the hypothesis that elective delayed sternal closure reduces the morbidity after arterial switch in neonates but does, however, confirm the safety and efficacy of the procedure. PMID- 11383800 TI - Tetralogy of Fallot: surgical management individualized to the patient. AB - BACKGROUND: Over the past four decades, the surgical trend has been toward early, complete repair of tetralogy of Fallot (TOF). Many centers currently promote all neonates for total correction irrespective of anatomy and symptoms, with some surgeons advocating hypothermic circulatory arrest for repair in small infants. We believe this approach increases morbidity. METHODS: Based on approximately 40 years' experience in 2,175 patients, we developed a management protocol focused on patient size, systemic arterial saturations, and anatomy. Symptomatic patients (hypercyanotic spells, ductal dependent pulmonary circulation) weighing less than 4 kg undergo palliative modified Blalock-Taussig shunt (BTS) followed by complete repair at 6 to 12 months. Asymptomatic patients, weighing less than 4 kg who have threatened pulmonary artery isolation, undergo BTS and repair at 6 to 12 months. All other patients undergo complete repair after 6 months. RESULTS: From July 1, 1995, to December 1, 1999, 144 patients underwent operation for TOF (129 patients) or TOF with atrioventricular septal defect (TOF/AVSD, 15 patients). Ninety-four patients underwent one stage complete repair (88 TOF, 6 TOF/AVSD). Thirty-nine patients underwent repair after initial BTS (32 TOF, 7 TOF/AVSD). Ten patients are awaiting repair after BTS. The mean age and weight at complete repair were 18 months and 9 kg. There were no operative deaths. There have been 3 late deaths with complete follow-up (mortality 3 of 144 [2.1%]). Four of 133 patients (3%) have required reoperation after total correction. CONCLUSIONS: This management strategy optimizes outcomes by individualizing the operation to the patient. Advantages include avoidance of circulatory arrest, low morbidity and mortality, and low incidence of reoperation after complete repair. PMID- 11383801 TI - Aortic valvotomy for congenital valvular aortic stenosis: a 37-year experience. AB - BACKGROUND: The purpose of the study was to analyze the long-term results of aortic valvotomy and the risk factors associated with reoperation and survival. METHODS: From 1960 to 1977, 116 patients with congenital valvular aortic stenosis underwent isolated aortic valvotomy at a mean age of 13.7 +/- 7.8 years with a mean aortic gradient of 78 +/- 33 mm Hg. Fifteen patients had additional aortic regurgitation, and leaflet calcification was present in another 15 patients. RESULTS: Postoperatively the mean aortic gradient decreased to 19.4 +/- 11.3 mm Hg (p < 0.0001). Early mortality was 2.6%. At a mean follow-up of 23.8 years, 26 late deaths (22.4%) occurred among the 113 early survivors. Actuarial 10-, 20-, 30-, and 37-year survival rates were 94.6%, 79.7%, 76.2%, and 72.5%, respectively. According to multivariate Cox regression analysis, survival was influenced by preoperative New York Heart Association class (p = 0.0418), leaflet calcification (p = 0.0339), date of operation (p = 0.0253), and postoperative endocarditis (p < 0.0001). At a mean interval of 18.3 years, 37 patients required reoperation (31.9%) mainly because of recurrent aortic stenosis. The reoperation rate increased significantly 15 years postoperatively from 0.73%/year to 2.31%/ year (p < 0.0001). In a multivariate risk model, reoperation was influenced by older patient age (p = 0.0032) and the presence of leaflet calcification (p = 0.0289). CONCLUSIONS: AORTIC valvotomy is a simple and effective procedure for congenital aortic stenosis with excellent long-term results. However, the rate of reoperation increases 15 years postoperatively, and clinical follow-up should be intensified. Our results suggest that early repair should be performed and that adequate patient selection is the most important determinant of the longterm results. PMID- 11383802 TI - Intranasal mupirocin reduces sternal wound infection after open heart surgery in diabetics and nondiabetics. AB - BACKGROUND: This study was designed to determine whether decreasing nasal bacterial colonization by applying Mupirocin (MPN) intranasally decreases sternal wound infections. METHODS: We prospectively followed 992 consecutive open heart surgery (OHS) patients who did not receive MPN prophylaxis (group I) from January 1, 1995 to October 31, 1996. Group II consisted of 854 consecutive patients followed prospectively from December 1, 1997 to March 31, 1999 treated with intranasal MPN given on the evening before, the morning of OHS, and twice daily for 5 days postoperatively. RESULTS: There was a significant difference in the rate of overall sternal wound infections between the untreated (group I) and the treated group (group II): 2.7% (27 of 992) versus 0.9% (8 of 854) (p = 0.005). The difference was also significant in the diabetic subgroup: 5.1% (14 of 277) (group I) versus 1.9% (5 of 266) (group II) (p = 0.04) and the nondiabetic group: 1.8% (13 of 715) (group I) versus 0.5% (3 of 588) (group II) (p = 0.03). The cost of MPN treatment was $12.47 per patient compared with $81,018 +/- $41,567 for a deep wound infection with no antibiotic-related complications recorded. CONCLUSIONS: Prophylactic intranasal MPN is safe, inexpensive, and very effective in reducing the overall sternal wound infections by 66.6%. PMID- 11383803 TI - Closed drainage using redon catheters for poststernotomy mediastinitis: results and risk factors for adverse outcome. AB - BACKGROUND: Several different surgical techniques have been described for the treatment of poststernotomy mediastinitis. The present study was undertaken to evaluate the midterm results of primary closed drainage using Redon catheters and to identify risk factors for adverse outcome. METHODS: Hospital records of 72 patients in whom poststernotomy mediastinitis developed and who underwent closed drainage with Redon catheters between April 1, 1996, and December 31, 1999, were reviewed. Follow-up was complete and averaged 11.8 +/- 11.5 months. RESULTS: Of the 25 deaths (34.7%) recorded, 15 were directly attributable to mediastinitis. Actuarial estimates for freedom from mediastinitis-related death were 80.1% at 1 month and 77.4% at 1 year, 2 years, and 3 years. Logistic regression identified older age (odds ratio, 1.1; 95% confidence interval, 1.02 to 1.18), incubation time of 14 days or less (6.5; 1.33 to 31.4), and methicillin-resistant Staphylococcus aureus (5.8; 1.2 to 27.2) as independent risk factors for mediastinitis-related death. Reintervention for recurrent mediastinitis was necessary in 9 patients (12.5%) and occurred at a mean interval of 18.7 +/- 13.5 days from the first debridement. Actuarial estimates for freedom from reintervention were 87.1% at 1 month and 85.2% at 1 year, 2 years, and 3 years. The combined end point of treatment failure (mediastinitis-related death or reintervention) was recorded in 9 patients (26.4%). Actuarial estimates for freedom from treatment failure were 74.3% at 1 month and 72.7% at 1 year, 2 years, and 3 years. Logistic regression identified older age (1.01; 1.02 to 1.18), preoperative renal insufficiency (6.8; 1.04 to 44.5), and methicillin resistant S aureus infection (4.8; 1.04 to 22.33) as independent risk factors for treatment failure (includes mediastinitis-related death and reintervention [with or without death]). CONCLUSIONS: Primary closed drainage using Redon catheters is an effective and simple treatment for most patients in whom poststernotomy mediastinitis develops. However, patients with methicillin-resistant S aureus infection or recurrent mediastinitis may benefit from a more aggressive approach. PMID- 11383804 TI - Cardiac hydatid cysts with intracavitary expansion. AB - BACKGROUND: Hydatid cyst disease is a significant health problem for undeveloped and developing countries. Although cardiac involvement is rare, early diagnosis and treatment of this situation is important. METHODS: To investigate the long term outcome of patients who underwent operation for cardiac hydatid cysts with intracavitary expansion, we reviewed 8 patients who had cardiac hydatidosis and who underwent operation in our institution between January 1988 and November 1999. All patients presented with intracavitary protrusion of the cysts. Seven patients were women. The mean age was 33 +/- 14.3 years with a range of 17 to 55 years. The cysts were located on the right ventricular outflow tract (2 patients), right midventricular part of the muscular septum, left atrial free wall and apical portions of the right (2), or left (2 patients) ventricle. Standard cardiopulmonary bypass and crystalloid antegrade cardioplegia with aortic cross-clamping were used in all patients. In one, with right ventricular hydatid cyst, we used cardiopulmonary bypass with femoral cannulation and total circulatory arrest at less than 18 degrees C systemic hypothermia. This patient, who was arrested because of pulmonary emboli could not be weaned from cardiopulmonary bypass and died. RESULTS: The cystic cavity was cleaned and closed with multiple pursestring sutures in 4 patients. In 2, cardiac and cystic cavities were united by partially resecting part of the cyst facing the cavity. In another patient, a left ventricular patch plasty was performed after removal of the cystic material in the left ventricle. Mebendazole was used postoperatively in all patients. Except for 1 patient who died, all were discharged without postoperative complications. The mean follow-up was 7.5 +/- 5 years. There was no late cardiac mortality or recurrence. CONCLUSIONS: Cardiac hydatid cysts with intracavitary expansion should be treated surgically without delay. Gentle handling of the heart during cardiopulmonary bypass minimizes operative risk. All patients should be investigated for systemic cysts. PMID- 11383805 TI - Protection of the spinal cord with pentobarbital and hypothermia. AB - BACKGROUND: Ischemic spinal cord damage during thoracic aortic operations has not been eliminated despite application of various adjuncts. We experimentally investigated the protective effects of pentobarbital and hypothermia on the spinal cord subjected to ischemia. METHODS: Among nine groups of 6 rabbits each, groups AI to AIII underwent 20-minute infrarenal aortic occlusion, and groups BI to BVI underwent 40-minute occlusion. Five milligrams per kilogram of pentobarbital was administered to groups AII and BII; 10 mg/kg in groups AIII, BIII, and BVI; 20 mg/kg in group BIV; and none in groups AI, BI, and BV. In groups BV and BVI, hypothermia was induced. Forty-eight hours postoperatively, the motor function of the lower limbs was evaluated. RESULTS: Statistically significant recovery of motor function was observed in animals in groups AII, AIII, BIII, BIV, BV, and BVI. CONCLUSIONS: Pentobarbital showed dose-dependent protective effects of the spinal cord. Moderate hypothermia alone also showed protective effects. Combined use of pentobarbital and hypothermia resulted in highly significant recovery of spinal cord function. PMID- 11383806 TI - Are macrophages involved in early myocardial reperfusion injury? AB - BACKGROUND: Neutrophils are the predominant phagocytes in the early stages of myocardial ischemia-reperfusion response and are also implicated in the development of tissue damage. This study examined the role of recruited macrophages in the evolution of this tissue injury. METHODS: Farm pigs were subjected to 30 minutes of myocardial ischemia followed by 30 minutes of reperfusion. Biopsy samples were taken from the control, ischemic, and ischemic reperfused left ventricle wall and processed for both morphologic and biochemical analyses. In situ production of tumor necrosis factor-alpha was evaluated by Western blot and immunofluorescence. A full hemodynamic evaluation was also performed. RESULTS: Myocardial ischemia and early reperfusion caused marked neutrophil and macrophage tissue accumulation and tumor necrosis factor-alpha production by the injured tissue. Immunofluorescence studies allowed us to localize tumor necrosis factor-alpha predominantly in tissue-infiltrating macrophages. No depression in the global myocardial contractile function was observed, either during ischemia or after reperfusion. CONCLUSIONS: These data suggest that the newly recruited macrophages within the ischemic and early post ischemic myocardium may play a role in promoting neutrophil tissue infiltration and subsequent neutrophil-induced tissue dysfunction by producing tumor necrosis factor-alpha. PMID- 11383807 TI - Efficacy of a new coating material, PMEA, for cardiopulmonary bypass circuits in a porcine model. AB - BACKGROUND: A new coating material, poly-2-methoxyethyl acrylate (PMEA), was developed to improve the biocompatibility of cardiopulmonary bypass (CPB) circuits. METHODS: To investigate the efficacy of the PMEA coating for CPB circuits, we compared PMEA-coated circuits (group P, n = 6) with uncoated circuits (group C, n = 6) and heparin (covalent-bonded heparin, Hepaface)-coated circuits (group H, n = 6) in a porcine CPB model. RESULTS: Platelet counts were significantly preserved in groups P and H compared with those in group C (P versus C, p < 0.05). The plasma levels of thrombin-antithrombin complex and bradykinin were significantly lower at 120 minutes in groups P and H than in group C (thrombin-antithrombin: P versus C, p < 0.05; bradykinin: P versus C, p < 0.05). The amount of fibrinogen adsorbed onto the hollow fibers was markedly less in group P than in groups C and H. CONCLUSIONS: The PMEA coating was equal to heparin coating in preventing reactions induced by CPB circuits, and might be superior to heparin coating in suppressing the adsorption of plasma proteins such as fibrinogen. Thus, PMEA coating may be a suitable means for improving the biocompatibility of CPB circuits. PMID- 11383809 TI - Prospective randomized trial compares suction versus water seal for air leaks. AB - BACKGROUND: Surgeons treat air leaks differently. Our goal was to evaluate whether it is better to place chest tubes on suction or water seal for stopping air leaks after pulmonary surgery. A second goal was to evaluate a new classification system for air leaks that we developed. METHODS: Patients were prospectively randomized before surgery to receive suction or water seal to their chest tubes on postoperative day (POD) #2. Air leaks were described and quantified daily by a classification system and a leak meter. The air-leak meter scored leaks from 1 (least) to 7 (greatest). The group randomized to water seal stayed on water seal unless a pneumothorax developed. RESULTS: On POD #2, 33 of 140 patients had an air leak. Eighteen patients had been preoperatively randomized to water seal and 15 to suction. Air leaks resolved in 12 (67%) of the water seal patients by the morning of POD #3. All 6 patients whose air leak did not stop had a leak that was 4/7 or greater (p < 0.0001) on the leak meter. Of the 15 patients randomized to suction, only 1 patient's air leak (7%) resolved by the morning of POD #3. The randomization aspect of the trial was ended and statistical analysis showed water seal was superior (p = 0.001). The remaining 14 patients were then placed to water seal and by the morning of POD #4, 13 patients' leaks had stopped. Of the 32 total patients placed to seal, 7 (22%) developed a pneumothorax and 6 of these 7 patients had leaks that were 4/7 or greater (p = 0.001). CONCLUSIONS: Placing chest tubes on water seal seems superior to wall suction for stopping air leaks after pulmonary resection. However, water seal does not stop expiratory leaks that are 4/7 or greater. Pneumothorax may occur when chest tubes are placed on seal with leaks this large. PMID- 11383808 TI - BioGlue surgical adhesive for thoracic aortic repair during coagulopathy: efficacy and histopathology. AB - BACKGROUND: We hypothesized that induction of coagulopathy in sheep would model clinical needle hole and surgical bleeding from synthetic graft anastomoses, and that a new tissue bioadhesive (BioGlue) would control postoperative blood loss during surgical repair of the thoracic aorta. METHODS: Sheep were anticoagulated with aspirin and heparin. A bypass was made using end-to-side anastomoses of a graft to a partially occluded descending thoracic aorta. Experimental anastomoses (EXP, n = 9) were treated with BioGlue, and control anastomoses (CON, n = 5) were treated with Surgicel to gain intraoperative hemostasis. RESULTS: EXP animals exhibited significantly reduced postsurgical bleeding (CON median 955 mL versus EXP median 470 mL, p < 0.003), a reduced rate of blood loss over the first 2 postoperative hours (CON median 210 mL/hr versus EXP median 92.5 mL/hr, p < 0.006), and over the entire recovery period (CON median 158 mL/hr versus EXP median 86 mL/hr, p < 0.05), and reduced total blood loss (CON mean 1,497 +/- 691 mL versus EXP mean 668 +/- 285 mL, p < 0.008). On histologic examination of tissues explanted after 3 months, BioGlue was relatively inert and demonstrated a minimal inflammatory response. CONCLUSIONS: The use of BioGlue significantly reduced the volume and rate of postsurgical bleeding in a coagulopathic sheep model for thoracic aortic operations. Histopathologically, BioGlue generated only a minimal inflammatory response. This new surgical tissue bioadhesive should prove extremely beneficial for coagulopathic patients undergoing thoracic aortic or vascular procedures. PMID- 11383810 TI - Randomized controlled trial of a synthetic sealant for preventing alveolar air leaks after lobectomy. AB - BACKGROUND: The intraoperative application of synthetic surgical lung sealant (SLS) to surfaces leaking air or at risk of air leaks has been advocated to reduce alveolar air leaks (AAL) after lobectomy. METHODS: This study was designed to investigate the effectiveness of SLS in reducing AAL in patients considered intraoperatively to have moderate to severe AAL, after all conventional measures to reduce such leaks had been used. Over 17 months, 124 patients undergoing standard lobectomy were randomized to standard closure of parenchymal surgical sites, with or without SLS. RESULTS: In treated patients, the mean numbers of intraoperative AAL after application of SLS were significantly smaller than in untreated patients (38.5 mL versus 59.9 mL, p = 0.0401). Postoperatively, the mean time to last observable AAL was shorter in the treated group (33.7 hours versus 63.2 hours, p = 0.0134) and the mean percentage of patients free of AAL at days 3 and 4 was smaller (87% versus 58.5%, p = 0.002). However, the occurrence of incomplete lung expansion after drain removal, and the length of the postoperative hospital stay due to prolonged AAL, were not different. In the treatment group, 4 patients developed localized empyema and incomplete lung expansion without bronchopleural fistula 7, 12, 15, and 20 days, respectively, after operation. In these 4 patients, inserted chest tubes drained infected sealant. CONCLUSIONS: Surgical lung sealant may be a useful adjunct to conventional techniques for reducing moderate and severe AAL after lobectomy, but its use seems to increase the risk of postoperative empyema. PMID- 11383812 TI - Endotracheal balloon dilatation and stent implantation in benign stenoses. AB - BACKGROUND: Tracheal reconstruction is the treatment of choice in nontumorous tracheal stenoses, but recurrences and concomitant medical conditions limit this approach. We investigated the outcome after balloon dilatation and silicone stent implantation. METHODS: Forty-two patients with inoperable tracheal stenoses underwent balloon dilatation and afterward silicone stent implantation. Patients were divided into two groups, in group A 24 patients received tracheal stents as a temporary treatment. In group B, definitive stenting was done in 18 patients with severe concomitant medical conditions that did not allow for stent removal. RESULTS: Immediate results were satisfactory in all patients. In group A, stents could be removed in 12 patients after a mean interval of 20 months. Restenting was not required during the following 18.9 months. Twelve patients are still waiting for stent removal after a mean follow-up of 20 months. In group B, mean follow-up is now 48.4 months. Complications included retained secretions, dislocation, and granuloma formation. CONCLUSIONS: Stenting after balloon dilatation is safe and effective in benign tracheal stenoses. After temporary use, stents can be removed when local and general conditions permit. In all other patients, stenting proved beneficial for 5 years as more definitive treatment. PMID- 11383811 TI - Trial of a novel synthetic sealant in preventing air leaks after lung resection. AB - BACKGROUND: Postoperative air leaks are a major cause of morbidity after lung resections. This study was designed to evaluate the efficacy and safety of a new synthetic, bioresorbable surgical sealant in preventing air leaks after pulmonary resection. METHODS: In a multicenter trial, 172 patients undergoing thoracotomy were randomized intraoperatively in a 2:1 ratio to receive surgical sealant applied to sites at risk for air leak after standard methods of lung closure (treatment group) or to have standard lung closure only (control group). The primary outcome variable was the percentage of patients free of air leakage throughout hospitalization. Secondary outcome variables were the control of air leaks intraoperatively and the time to postoperative air leak cessation. Time to chest tube removal, time to hospital discharge, and safety outcomes were also evaluated. RESULTS: Air leaks were identified before randomization in 89 of 117 patients in the treatment group and in 39 of 55 patients in the control group. Application of the sealant resulted in control of air leaks in 92% of treated patients (p < or = 0.001). A significantly higher percentage of treated patients than control patients remained free of air leaks during hospitalization (39% versus 11%, p < or =0.001). The mean times to last observable air leak were 30.9 hours in the treatment group and 52.3 hours in the control group (p = 0.006). In the treatment group, trends were observed for reduced time to chest tube removal and earlier discharge. No significant difference was identified in postoperative morbidity and mortality between the two groups. CONCLUSIONS: Air leaks after lung resection occur in most patients. The application of this novel surgical sealant appears to be effective and safe in preventing postoperative air leaks. PMID- 11383813 TI - Expression of vascular endothelial growth factor in thoracic sarcomas. AB - BACKGROUND: A body of data indicates that vascular endothelial growth factor (VEGF) expression by carcinomas is closely related to the prognosis of carcinomas. However, the relationship between VEGF expression and the prognosis of sarcomas is contradictory. METHODS: Tissue from 27 cases of thoracic sarcoma was analyzed immunohistochemically for VEGF expression while tumor vascularity was quantified using an antibody directed against endothelial CD34. The relationship between VEGF expression and the prognosis of patients with sarcomas was then evaluated semiquantitatively. RESULTS: The microvessel count in sarcomas with strong VEGF expression was significantly higher than that in sarcomas with absent or faint VEGF expression. The disease-free survival rates of sarcomas with strong VEGF expression were significantly lower than those of sarcomas with absent or faint VEGF expression. We found that strong VEGF expression impacted on the disease-free survival in multivariate analyses. CONCLUSIONS: VEGF expression of thoracic sarcomas is directly related to angiogenesis and tumor vascularity, and our findings suggest that strong VEGF expression is an independent prognostic factor in patients with thoracic sarcomas. PMID- 11383814 TI - Evolving management and outcome of esophageal cancer with airway involvement. AB - BACKGROUND: Esophageal cancer with airway involvement, including patients with esophagorespiratory fistula (ERF), has been associated with a poor prognosis. Multimodality treatment, self-expanding metal stents, and improved supportive therapy may be impacting outcome in these patients. There is concern for the development of ERF during therapy. METHODS: We retrospectively studied 74 consecutive male patients at a single institution presenting between 1/85 to 12/98 with bronchoscopic, endoscopic or radiographic confirmation of airway involvement with esophageal cancer, including 35 patients with ERF. Comparison was made between the first 35 patients (group I) and the last 39 patients (group II) with regard to antineoplastic therapy, stent placement, and survival. RESULTS: Treatment in group I included supportive care in 17 of 35 patients, plastic stent in 7 of 35 patients, and radiation or chemotherapy in 9 of 35 patients. In group II, radiation or chemotherapy was offered to 33 patients, and self-expanding metal stents were placed in 10 of 39 patients. Surgical resection was possible after neoadjuvant therapy in 13 of 39 patients in group II, including 2 initially presenting with ERF. Median survival in group I was 16 weeks and in group II was 37 weeks. Comparison of Kaplan-Meier survival estimates using log rank testing demonstrated improved survival in group II (p = 0.0026). Long-term survival in 4 group II patients initially presenting with ERF and receiving multimodality treatment was observed. Development of ERF during treatment occurred in 3 group II patients. Treatment failure was predominantly local in group I and local and distant in group II. CONCLUSIONS: More aggressive treatment may favorably influence outcome in esophageal cancer with airway invasion. Long-term survival and the development of ERF during therapy occurred at similar rates. PMID- 11383815 TI - Retinoic acid enhances lung growth after pneumonectomy. AB - BACKGROUND: We sought to identify the role of retinoic acid (RA) upon lung growth. RA has a role in perinatal lung development, and we hypothesized that exogenous RA would enhance postpneumonectomy compensatory lung growth. METHODS: Utilizing the postpneumonectomy rat model, we studied the impact of RA upon contralateral lung growth. Adult Sprague-Dawley rats were divided into three groups. Group S underwent a sham left thoracotomy, group P underwent left pneumonectomy, and group R underwent left pneumonectomy with administration of exogenous RA (0.5 microg/g/day intraperitoneally). We then quantitated right lung growth after 10 and 21 days. Lung weight and volume were expressed as a ratio to the final body weight (lung weight and volume indices, LWI and LVI). Epidermal growth factor receptor (EGFR) expression was quantitated using Western blot analysis. Cellular proliferation index (CPI) was determined using BrdU immunostaining. RESULTS: LWI, LVI, CPI, and EGFR expression at 21 days were significantly higher in group R versus S and P. At the 10-day interval, both LWI and LVI were significantly higher in group R versus S and P. CONCLUSIONS: RA administration markedly enhances lung growth after pneumonectomy, as evidenced by increases in LWI, LVI, and CPI. Upregulation of EGFR expression was associated with these effects. PMID- 11383816 TI - Recipient intramuscular gene transfer of active transforming growth factor-beta1 attenuates acute lung rejection. AB - BACKGROUND: Gene transfer into the donor graft has been demonstrated to be feasible in reducing ischemia-reperfusion injury and rejection in lung transplantation. This study was undertaken to determine whether intramuscular gene transfer into the recipient can also reduce subsequent lung graft rejection. METHODS: Brown Norway rats served as donors and F344 rats as recipients. Recipient animals were injected with 10(10) plaque-forming units of adenovirus encoding active transforming growth factor beta1 (group I, n = 6), beta galactosidase as adenoviral controls (group II, n = 6), or normal saline without adenovirus (group III, n = 6) into both gluteus muscles 2 days before transplantation. Gene expression was confirmed by enzyme-linked immunosorbent assay. Graft function was assessed on postoperative day 5. RESULTS: Successful gene transfection and expression were confirmed by the presence of active transforming growth factor beta1 protein in muscle and plasma. Oxygenation was significantly improved in group I (group I vs II and III, 353.6 +/- 63.0 mm Hg vs 165.7 +/- 39.9 and 119.1 +/- 41.5 mm Hg; p = 0.02 and 0.004). The muscle transfected with the transforming growth factor beta1 showed granulation tissue with fibroblast accumulation. CONCLUSIONS: Intramuscular adenovirus-mediated gene transfer of active transforming growth factor beta1 into the recipients attenuates acute lung rejection as manifested by significantly improved oxygenation in transplanted lung allografts. This intramuscular transfection approach as a cytokine therapy is feasible in transplantation and may be useful in reducing rejection as well as reperfusion injury. PMID- 11383817 TI - Combretastatin A-4 prodrug inhibits growth of human non-small cell lung cancer in a murine xenotransplant model. AB - BACKGROUND: Combretastatin A-4 prodrug (CA-4PD) has been identified as a potent antivascular agent in various rodent tumor models. The aim of this study was to investigate the effect of CA-4PD on human non-small cell lung cancer (NSCLC). METHODS: Cytostatic and cytotoxic effects of CA-4PD on selected NSCLC cells, Colo 699 and KNS-62, were studied in vitro. After subcutaneous xenotransplantation the effect of systemically administrated CA-4PD on tumor growth was investigated in vivo. A newly established orthotopic xenotransplant model was employed to estimate prolongation of survival after intrapulmonary tumor induction with secondary metastatic disease. RESULTS: In vitro, CA-4PD displayed a time and dose dependent antiproliferative effect on human lung cancer cells. In vivo, CA-4PD significantly delayed growth of subcutaneously induced lung cancer. This growth delay was translated into a prolongation of survival in the metastasizing orthotopic xenotransplant model. CONCLUSIONS: In vitro CA-4PD inhibits proliferation of NSCLC cells, most likely by disruption of microtubule assembly. In vivo, systemic treatment inhibits growth of subcutaneously xenotransplanted tumors by an antivascular effect. In the case of metastasizing human lung cancer this translated into a prolongation of survival. PMID- 11383818 TI - Tracheal allotransplantation maintaining cartilage viability with long-term cryopreserved allografts. AB - BACKGROUND: Cartilage viability of a cryopreserved tracheal allograft seems to affect graft function and durability. We previously reported the influence of warm ischemia and cryopreservation on cartilage viability of tracheal allografts. For the clinical application of tracheal allotransplantation, it is essential to preserve grafts for a long time. In this study, we assessed cartilage viability of tracheal allografts after long-term cryopreservation in transplantation models. METHODS: The tracheas were harvested from Lewis rats. The grafts were frozen to -80 degrees C in a programmable freezer immediately after being harvested and were then stored in liquid nitrogen (-196 degrees C) for different lengths of preservation (1, 2, 6, 9, 12, 18, and 24 months; n for each group = 8). Cartilage viability was evaluated by estimating proteoglycan synthesis. After harvest or thawing of the tracheas, the cartilage was labeled with 4 muCi/mL of Na2 35SO4. Specimens were then hydrolyzed in 0.5 mol/L NaOH, and a solution of the extracts was then counted by a liquid scintillation counter. 35Sulfur incorporation before and after cryopreservation was examined in each group. Tracheal allotransplantation was performed using Lewis rats as donors and Brown Norway rats as recipients. RESULTS: The average 35S incorporation in the cartilage before cryopreservation was 224 +/- 17 disintegrations per minute per milligram of tissue protein. The average 35S incorporation in the cartilage after cryopreservation decreased to 67% to 76% compared with that before cryopreservation. There were no significant differences among the groups in 35S incorporations after cryopreservation. Histologic examination after transplantation revealed normal tracheal cartilage in all groups. CONCLUSIONS: The viability of tracheal cartilage after cryopreservation decreased to 67% to 76%. There were no significant differences in viability of cartilage among the tracheas after different lengths of cryopreservation. Tracheal allotransplantation after long-term cryopreservation can be safely performed in the rat model. PMID- 11383819 TI - Dissection of atrial septum after mitral valve replacement. AB - We report a patient who presented with paraprosthetic leak complicated by dissection of the interatrial septum after mitral valve replacement. A review of the literature provides confirmation that only 3 cases have been previously reported of this potential, albeit extremely rare, complication of prosthetic mitral valve replacement. Prosthesis oversizing and improper mitral annular handling appeared to be the predisposing factors of this complication. PMID- 11383820 TI - Three-year follow-up of an atrial septal aneurysm. AB - This is a 3-year follow-up of a gradually ballooning atrial septal aneurysm (ASA) which developed a spontaneous echo contrast and later a mobile thrombus in the aneurysm. This clearly demonstrates one of the pathogenetic mechanisms of systemic thromboembolism associated with ASA. In view of the risk of systemic thromboembolism or the need for lifelong anticoagulation treatment, the aneurysm was excised and an atrial septal patch was fashioned to close the resultant defect. PMID- 11383821 TI - Left ventricular hamartoma associated with ventricular tachycardia. AB - Cardiac hamartomas are a rare type of benign tumor affecting the heart. We describe a 33-year-old patient who presented with a wide complex tachycardia. Diagnostic imaging revealed a mass in the patient's left ventricular wall, near the apex of the heart. The mass was surgically resected and appeared benign. Its pathology was that of a hamartoma of mature cardiac myocytes. Postoperative electrophysiology evaluation showed no inducible focus and the patient remains alive and asymptomatic after 2 years of follow-up. PMID- 11383822 TI - A primary cardiac sarcoma with unusual histology and clinical course. AB - A left atrial tumor, in which radical resection was impossible, demonstrated two processes: An inflammatory pseudotumor and cellular atypia suggestive of a sarcoma. Immunohistochemistry (proliferating cell nuclear antigen [PCNA], MIB-1 [Ki-67 antibody], bcl-2 positive; p53 negative, focal loss of nm23) was supportive for a malignant tumor. Despite no further therapy because of uncertainty in tumor classification, the patient remained in remission for 28 months. Thereafter, spine metastases and local regrowth were found, and the patient died 15 months later, after temporary remission by radiotherapy. This case stresses the impact immunohistochemistry may have on diagnosis of malignancy and the difficulty in predicting the biological behavior of cardiac sarcomas. PMID- 11383823 TI - Cardiac papillary fibroelastoma on the pulmonary valve: a rare cardiac tumor. okaken@mcai.med.hiroshima-u.ac.jp. AB - We report the case of a patient with a rare papillary fibroelastoma on the pulmonary valve visualized before surgical intervention. The tumor was an encapsulated, rounded mass. The gelatinous membrane on the surface tore easily, and multiple fronds appeared. This case emphasizes that when there is an encapsulated mass attached to a valve, the initial excision of valve tissue should be as minimal as possible. To avoid unnecessary injury to the valve, it is simple and practical to confirm that the tumor has the appearance of a sea anemone, thus identifying it as a papillary fibroblastoma, a benign tumor. PMID- 11383824 TI - Safe technique for removal of extensive renal cell tumors. AB - We describe our technique for excision of a renal tumor that extended to the right atrium and was adherent to the inferior vena cava within the liver. The surgical procedure was performed using cardiopulmonary bypass with deep hypothermic circulatory arrest and retrograde cerebral perfusion to extend the safe period of cerebral ischemia. PMID- 11383825 TI - Successful VATS ligation of a large anomalous branch producing IMA steal syndrome after MIDCAB. AB - The occurrence of a flow "steal phenomenon" from a large branch of the internal mammary artery (IMA) is rare and its pathogenesis is still controversial. We describe a case of a patent large anomalous left IMA branch which produced recurrent angina 3 years post MIDCAB. Transcatheter coil obliteration of the vessel produced symptomatic relief. However, six months later, chest pains recurred in association with unwinding of the coil and recannulization of the collateral vessel. Successful ligation of the culprit branch using a VATS approach is presented. PMID- 11383826 TI - Intravascular migration of fractured sternal wire presenting with hemoptysis. AB - We present a rare complication of median sternotomy in which a segment of fractured sternal wire punctured the heart, embolized to the right lung, and eroded into a bronchus causing massive hemoptysis. It was safely removed through a median sternotomy. Sternal wire fracture or migration is diagnosed easily on chest roentgenograms, but frequently goes unnoticed. Sternal wire failure can be managed nonoperatively; however, repair is indicated if fractured wires are displaced or potentially migratory. PMID- 11383827 TI - Fenestrated arterial switch operation: surgical approach to an unusual transposition of the great arteries complex. AB - Transposition of the great arteries with intact ventricular septum and aortopulmonary window is an extremely rare anatomic combination, having been reported just twice previously. Other authors performed a physiologic repair, because the combination was considered unsuitable for an anatomic repair. We describe the case of a 26-day-old baby with such anatomy who was successfully treated with an arterial switch operation. A 4 mm fenestration at atrial level was made for a smoother postoperative course. PMID- 11383828 TI - Transposition with absent pulmonary valve syndrome: early repair of a rare case. AB - The congenital absence of the pulmonary valve cusps can occur either isolated or in association with other heart lesions. We report a very rare case of a 40-day old infant with transposition of the great arteries, ventricular septal defect, pulmonary annular stenosis, absent pulmonary valve and aneurysmal dilation of the central pulmonary arteries, who received surgical treatment at our institution. PMID- 11383829 TI - Idiopathic main pulmonary artery aneurysm. AB - Main pulmonary artery aneurysms with no underlying pathology are very rare, and the role of surgery in this entity is not well defined. We report our experience of one case. PMID- 11383830 TI - Helical computed tomographic angiography in obstructed total anomalous pulmonary venous drainage. AB - Helical computed tomographic angiography with differential color imaging technique clearly demonstrated pulmonary venous obstruction in an infant with total anomalous pulmonary venous drainage before and after operation. This technique is less invasive and provides precise spatial information of complicated vascular anomalies. PMID- 11383831 TI - Giant midesophageal pulsion diverticulum: a report of two operated cases. AB - A giant midesophageal pulsion diverticulum is a medical rarity. Two successfully operated cases are reported. One patient had no clinical symptoms and was misinterpreted as a mediastinal tumor. Esophageal myotomy was not performed. PMID- 11383832 TI - Metalloptysis: a late complication of lung volume reduction surgery. AB - We describe three cases where patients expectorated titanium staples many months after lung volume reduction surgery (LVRS). The possible mechanisms and technical implications of this rare complication are discussed. PMID- 11383833 TI - Redoing reconstruction of the esophagus using remnants of the ileo-left colon aided by microvascular anastomosis. AB - Theoretically, the jejunum, fasciocutaneous or myocutaneous flap is recommended as an esophageal substitute in redoing reconstruction of the esophagus after a second incidence of corrosive injury. However, other esophageal substitutes should also be considered. We present a case of a 42-year-old woman who underwent esophageal reconstruction using an ileocolon graft for corrosive esophageal stricture ten years before. The patient ingested caustic drain cleaner again and underwent resection of the ileocolon graft secondary to corrosive necrosis. Two and a half months after the second incidence of corrosive injury, reconstruction of the esophagus was again performed using a graft of remnant ileo-left colon aided by microvascular anastomosis. The patient was able to swallow a regular diet after the procedure. Remnant ileo-left colon is a good alternative esophageal substitute in cases of repeated corrosive injury. PMID- 11383834 TI - Thoracoscopic resection of an ectopic intrathoracic goiter. AB - We report a case of an ectopic intrathoracic goiter in a 79-year-old human. This uncommon finding presented as a symptomatic paratracheal mass that was resected using thoracoscopic techniques without complication. PMID- 11383835 TI - Intrathoracic desmoid tumor mimicking primary lung neoplasm. AB - Most reported thoracic desmoid tumors originate from the chest wall. However, intrathoracic desmoid tumors are rare. The pathogenesis of these tumors is unclear but antecedent trauma and operation have been implicated. Desmoid tumors can present either with pain or be incidentally detected on radiographic studies. We describe the case of a 60-year-old woman with an intrathoracic desmoid detected on a routine screening chest roentgenogram who underwent complete surgical resection of the tumor. PMID- 11383836 TI - Bronchial anastomotic stricutre caused by ossification of an intercostal muscle flap. AB - We report a case of heterotopic ossification of a pedicled intercostal muscle flap that had been wrapped circumferentially around a bronchial sleeve anastomosis. This ossification caused severe bronchial stenosis and recurrent pneumonias. Stent insertion failed, and the patient ultimately required completion pneumonectomy. We recommended that caution be used when wrapping intercostal muscle around any important lumen. PMID- 11383837 TI - A unique congenital mediastinal malformation. AB - We report a case of a 35-year-old patient presenting with a unique asymptomatic malformation associating extralobar pulmonary sequestration communicating with a bronchogenic cyst of the esophageal wall via the aortopulmonary window, dextroisomerism, and complete agenesia of the left pericardium. Despite computed tomography (CT) scan and magnetic resonance imaging (MRI), the diagnosis could not be established before left thoracotomy. The sequestrated lobe and bronchogenic cyst were then successfully resected. PMID- 11383838 TI - Surgical correction of postpneumonectomy stridor by saline breast implantation. AB - Postpneumonectomy syndrome is a rare complication of pneumonectomy and is characterized by progressive dyspnea, stridor, and repeated chest infections. It is caused by displacement and rotation of the mediastinal structures into the pneumonectomy space, producing compression and malacic changes in the trachea and remaining bronchus. We report the successful long-term results of mediastinal correction, cardiopexy and plombage with saline breast prostheses in a 59-year old man after right pneumonectomy for carcinoma of the lung. PMID- 11383839 TI - Multislice helical computed tomography for minimally invasive cardiac surgery. PMID- 11383840 TI - Aortic stent-graft for patent ductus arteriosus in adults: the aortic exclusion technique. AB - The risk of closure of a patent ductus arteriosus in the elderly is high because of the fragility of the aorta and aneurysmal change in the ductus. Stent-grafting has emerged as a method of treatment of pathology of the aorta. We describe a case where this new endovascular technique has been successfully applied for closure of a patent ductus arteriosus in a high-risk patient. It may become part of the armamentarium for this pathology in adults. PMID- 11383841 TI - Surgical treatment of an aneurysm in the right aortic arch with aberrant left subclavian artery. AB - A saccular aneurysm in the right-sided aortic arch with aberrant left subclavian artery is an uncommon disease, and surgical treatment is complicated. Three patients with Edwards type III-B right aortic arch and enlargement of the Kommerell's diverticulum underwent operations. Right thoracotomy was the preferred approach for this lesion and partial cardiopulmonary bypass is a safe and simple procedure when the aortic arch has mild atherosclerosis. PMID- 11383842 TI - New method of coronary artery occlusion for beating-heart coronary artery bypass surgery. AB - Temporary occlusion of the coronary artery is an essential step in beating-heart coronary artery bypass surgery. The traditional method of occlusion of the coronary artery, by looping and suspension with thread or elastic slings, carries some potential risk for damage to the vessels. We report another method for temporary coronary artery occlusion involving parallel placement of stitches with pledgets along the coronary artery distal and proximal to the target area. This offers a clean operative field by gentle occlusion of the target coronary artery. PMID- 11383843 TI - The use of adenosine for repair of penetrating cardiac injuries: a novel method. AB - The use of intravenous administration of adenosine to expedite cardiorrhaphy in penetrating cardiac trauma by inducing temporary asystole is described. It is quicker, more effective, and safer than the traditional methods. PMID- 11383844 TI - Transxiphoid approach for intracardiac repair using video-assisted cardioscopy. AB - Video-assisted cardioscopy (VAC) is a novel tool for providing clear visualization of small intracardiac structures and achieving complete repair in minimally invasive surgery. Between July 1999 and July 2000, 12 patients with atrial septal defect and ventricular septal defect underwent surgical repair using a combined procedure with the transxiphoid approach and VAC in our institution. The mean skin incision was 5.4 cm, and the postoperative courses of all the patients were uneventful without any complications. The mean hospital stay was 8.3 days and 1 patient (8.3%) needed blood products. Our experience showed the technical feasibility and acceptable surgical results of transxiphoid approach using a VAC. PMID- 11383845 TI - A new off-pump technique for thoratec right ventricular assist device insertion. AB - The need for right ventricular support as an adjunct to left ventricular assistance is uncommon. When required, the insertion of a right ventricular assist device may be complicated by preexisting hepatic dysfunction, coagulation abnormalities, and renal failure, all of which are exacerbated by cardiopulmonary bypass. We report a technique for insertion of a right ventricular assist device without the need for cardiopulmonary bypass. PMID- 11383846 TI - Video-assisted extended thymectomy in patients with thymoma by lifting the sternum. AB - We present use of minimally invasive video thoracoscopic surgery to perform complete extended thymectomy in patients with thymoma. These procedures were performed using a sternum-elevating method that provides a wide field of vision between the sternum and heart. Indications for this method are Masaoka Stage I, II and some Stage III (invasion to the lung and pericardium). This new method may be useful from the standpoint of minimal access, rapid recovery, less pain, and good cosmetic results. PMID- 11383847 TI - Myocyte transplantation for myocardial repair: a few good cells can mend a broken heart. AB - Cell transplantation is a potential therapeutic approach for patients with chronic myocardial failure. Experimental transplantation of neonatal and fetal cardiac myocytes showed that the grafted cells can functionally integrate with and augment the function of the recipient heart. Clinical application of this approach will be limited by shortage of donors, chronic rejection, and because it is ethically contentious. By contrast skeletal myoblasts (satellite cells) are abundant and can be grafted successfully into the animal's own heart even after genetic manipulation in vitro. Functional integration of myoblasts, however, is hampered by the lack of intercellular gap junction communication and the difference in excitation-contraction coupling between skeletal and cardiac myocytes. In experimental studies several other cell types have been used to augment cardiac function. In this review we discuss the published results of myocyte transplantation with emphasis on potential sources of cells, the ethics of using donor embryonic and fetal cardiomyocytes, genetic transformation of skeletal myoblasts for myocardial repair, and the functional benefits of cell transplantation to the failing heart. PMID- 11383848 TI - As originally published in 1993: preoperative use of erythropoietin for cardiovascular operations in anemia. Updated in 2001. PMID- 11383849 TI - As originally published in 1993: skeletal muscle ventricles: left ventricular apex to aorta configuration. Updated in 2001. PMID- 11383850 TI - Antithrombotic treatment in patients with unstable coronary artery disease undergoing CABG. PMID- 11383851 TI - Remaining procoagulant property of wound blood washed by a cell-saving device. PMID- 11383852 TI - Efficacy of off-pump coronary artery bypass grafting in high-risk patients. PMID- 11383853 TI - OPCAB selection bias. PMID- 11383854 TI - Mitral valve repair in the elderly. PMID- 11383855 TI - Simple shunting of thoracoabdominal aneurysms with visceral perfusion. PMID- 11383856 TI - Successful recovery of allograft failure with biventricular support. PMID- 11383857 TI - Different technical approaches on cardiac binding procedures. PMID- 11383858 TI - Controlled perfusion of the transplanted lung. PMID- 11383859 TI - The potential of intratumoural unsealed radioactive source therapy. PMID- 11383860 TI - Biodistribution and dosimetry of 99mTc-ciprofloxacin, a promising agent for the diagnosis of bacterial infection. AB - This study reports on the biodistribution and dosimetry of technetium-99m ciprofloxacin, a radio-ligand developed for the visualisation of bacterial infection. Whole body scans were performed up to 24 h after intravenous injection of 370 MBq 99mTc-ciprofloxacin in three male and three female volunteers. Blood samples were taken at various times up to 24 h after injection. Urine was also collected up to 24 h after injection, allowing calculation of renal clearance and interpretation of whole body clearance. Time-activity curves were generated for the thyroid, heart, liver and whole body by fitting the organ-specific geometric mean counts, obtained from regions of interest. The MIRD formulation was applied to calculate the absorbed radiation doses for various organs. The images showed rapid, predominantly urinary excretion of 99mTc ciprofloxacin, with low to absent brain, lung and bone marrow uptake and low liver uptake and excretion. Accordingly, imaging conditions are excellent for both the thoracic and the abdominal region, even at early time points (60 min) post injection. In none of the volunteers was the gallbladder visualised. Approximately 60% of the injected activity was recovered in urine by 24 h post injection. The highest absorbed doses were received by the urinary bladder wall, the thyroid, the upper large intestine, the lower large intestine and the uterus. The estimated mean effective dose for the adult subject, taking into account the weight factors of the ICRP60 publication, was 0.0083 mSv/MBq. The amount of 99mTc ciprofloxacin required for adequate planar and tomographic imaging results in an acceptable effective dose to the patient. PMID- 11383861 TI - The value of 99mTc-HMPAO labelled white blood cell scintigraphy in acute appendicitis patients with an equivocal clinical presentation. AB - Various imaging studies can be performed in the evaluation of patients with a clinical presentation equivocal for acute appendicitis. One of these studies is technetium-99m hexamethylpropylene amine oxime (HMPAO) labelled white blood cell (WBC) scintigraphy. The aim of this study was to evaluate the accuracy and clinical value of 99mTc-HMPAO WBC scintigraphy in the aforementioned group of patients. Forty-one patients who had acute right lower quadrant abdominal pain with a clinical presentation equivocal for acute appendicitis were included in the study. The anterior abdomen and pelvis were imaged up to 4 h after the injection of 125-300 MBq 99mTc-HMPAO WBCs and the results were immediately reported to the surgeon before a decision was taken on whether to perform laparotomy. Diagnostic accuracy was established by the intra-operative findings and the histopathology in operated patients. In non-operated patients, absence of abdominal symptoms 1 month after scintigraphy and/or identification of another cause of abdominal pain was used to rule out acute appendicitis. There were 16 patients with positive scintigraphy and 81% of these patients were positive within 2 h post injection. There were no false-positive or false-negative results. We operated on 17 (41.4%) patients, and only one patient (5.9%) underwent unnecessary laparotomy. We conclude that 99mTc-HMPAO WBC scintigraphy is a rapid, highly accurate method for the exclusion of acute appendicitis and that its use can lower the unnecessarily high laparotomy rate in patients with an equivocal clinical presentation. PMID- 11383862 TI - Penile lymphoscintigraphy for sentinel node identification. AB - Lymphoscintigraphy for sentinel node (SN) identification has been extensively validated in breast cancer and melanoma. The aim of this study was to evaluate the findings of lymphoscintigraphy for SN identification in carcinoma of the penis. Lymphoscintigraphy was performed in 74 consecutive patients (mean age 62.2 years, range 28-87 years) with clinically lymph node-negative squamous cell carcinoma of the penis (stage T2 or greater). Following local anaesthesia by xylocaine 10% spray, technetium-99m nanocolloid (mean dose 64.8 MBq, range 40-131 MBq) in a volume of 0.3-0.4 ml was injected intradermally around the tumour. Shortly after injection, a 20-min dynamic study was performed with a dual-head gamma camera; subsequently, static anterior and lateral images were obtained at 30 min and 2 h using simultaneous cobalt-57 flood source transmission scanning. 57Co-assisted skin marking defined SN location for gamma probe/blue dye-guided biopsy, which was performed the next day. The SN visualization rate was 97% (72/74). Lymphatic drainage was bilateral in 81% of the cases (58/72), exclusively to the left groin in 13% (9/72) and only to the right groin in 6%. Bilateral lymph node drainage was synchronous in 38% (22/58) and asynchronous in 62% (in 18 patients the initial route was the left groin, and in the other 18, the right groin). Visualization before 30 min occurred in 66 patients (93%), in 64 of them (88%) already during the dynamic study. A total of 173 SNs were visualized (85 in the right groin, 88 in the left groin). Pitfalls were caused by inguinal skin contamination during injection (four patients) and intracavernous administration (one patient). At surgery, a total of 161 SNs were identified and removed. Sixteen patients (22%) had a tumour-positive SN and underwent standard regional lymph node dissection subsequently. During follow-up (median 28 months, range 3-74 months), two patients with a negative SN developed lymph node metastases in the mapped basin. It is concluded that penile lymphoscintigraphy is a valid and well-tolerated method for lymphatic mapping and SN identification. Although bilateral early inguinal drainage is the most frequent pattern, late imaging is recommended principally in patients with initial unilateral drainage in order to exclude delayed lymph node filling in the contralateral groin. SN identification may lead to a more accurate staging and avoid extensive lymph node dissection in the majority of patients with penile carcinoma. PMID- 11383863 TI - Analysis of FDG uptake with hybrid PET using standardised uptake values. AB - The standardised uptake value (SUV) has been used as an index of glucose metabolism to classify malignant tumours. To date, calculation of SUVs has been restricted to dedicated PET. The aim of this study was to investigate the feasibility of SUV calculation with attenuation-corrected hybrid PET, applying a singles count rate-related calibration method. Calibration factors for hybrid PET at different singles count rates were determined by phantom studies. SUVs were determined for hot spheres in a phantom study as well as for 68 malignant lesions in 56 patients. Recovery coefficients calculated for hot spheres were applied to SUVs of malignant lesions to correct for partial volume and recovery effects. At a sphere-to-background ratio of 10:1, SUVs of spheres with diameters from 34 to 16 mm varied from 5.0 to 1.5 for hybrid PET, and from 8.0 to 4.3 for dedicated PET. SUVs of malignant lesions calculated by hybrid and dedicated PET showed a strong correlation (r=0.95, P<0.001), with a mean percentage difference of 36%. SUVs calculated by hybrid PET were significantly lower than SUVs calculated by dedicated PET (6.2+/-4.3 vs 8.5+/-5.3, P<0.001). Application of recovery coefficients revealed an SUV of 12.2+/-7.3 for hybrid PET versus 10.8+/-6.3 for dedicated PET, with a significant reduction in the mean percentage difference (22%, P<0.01). In conclusion, singles count rate-related calibration factors allow calculation of SUVs with hybrid PET for lesions with a diameter larger than 15 mm. Correction for partial volume and recovery effects is needed to improve the agreement of SUVs of lesions determined by hybrid PET and dedicated PET. PMID- 11383864 TI - Assessment of left ventricular diastolic function from quantitative electrocardiographic-gated 99mTc-tetrofosmin myocardial SPET. AB - We have developed new software which can evaluate left ventricular (LV) diastolic functional parameters from a quantitative gated SPET (QGS) program. To examine its accuracy, we compared these findings with the LV diastolic functional indices obtained from gated radionuclide ventriculography (RNV). Twenty-four patients were selected for this study. Gated SPET with technetium-99m tetrofosmin was performed and the QGS program was used with a temporal resolution of 32 frames per R-R interval. The LV volume of each frame was calculated and four harmonics of Fourier series were retained for the analysis of the LV volume curve. From this fitted curve and its first derivative curve, we derived LV systolic functional indices, e.g. ejection fraction (EF), peak ejection rate (PER) and time to PER (TPER), as well as LV diastolic functional variables, e.g. 1/3 filling fraction (1/3 FF), peak filling rate (PFR) and time to PFR (TPFR). Within 5+/-2 days, gated RNV was performed and diastolic functional parameters were determined by the same method. No significant difference was observed between the variables calculated by gated SPET and by gated RNV. There was a good correlation between EF, PER, TPER, 1/3 FF, PFR and TPFR determined by these two methods (EF: r=0.95, P<0.0001; PER: r=0.87, P<0.0001; TPER: r=0.84, P<0.0001; 1/3 FF: r=0.87, P<0.0001; PFR: r=0.92, P<0.0001; TPFR: r=0.89, P<0.0001). Bland-Altman plots did not reveal any significant degree of directional measurement bias in any of the comparisons of gated SPET data and RNV data. It is concluded that, in addition to the conventional LV systolic functional indices, our program accurately provides LV diastolic functional parameters from gated SPET. Also, this program will be useful for detecting LV diastolic dysfunction in various cardiac diseases before LV systolic dysfunction becomes evident. PMID- 11383865 TI - Non-Q-wave myocardial infarction: impaired myocardial energy metabolism in regions with reduced 99mTc-MIBI accumulation. AB - Reduced regional technetium-99m methoxyisobutylisonitrile (99mTc-MIBI) accumulation in patients with chronic non-Q-wave infarction (NQWI) but without significant coronary artery stenosis indicates non-transmural damage of the myocardial wall. The aim of this study was to characterise cardiac energy metabolism after NQWI using phosphorus-31 magnetic resonance spectroscopy (31P MRS) and to compare the biochemical remodelling with changes in regional 99mTc MIBI uptake and with morphological and functional parameters assessed by magnetic resonance imaging (MRI). Fifteen patients with a history of NQWI, exclusion of significant coronary artery stenosis (<50% diameter stenosis) and hypokinesia of the anterior wall (group A) were examined with 31P-MRS to study the effects of NQWI on myocardial energy metabolism. Spectroscopic measurements were performed in the infarct-related myocardial region. Corresponding gradient-echo MR images and myocardial 99mTc-MIBI single-photon emission tomography images were acquired for exact localisation of the infarct region. All examinations were performed at rest under anti-ischaemic medication. Data were compared with those of patients in whom coronary artery disease had been excluded by angiography (group B, n=10). All patients of group A displayed anterior wall hypokinesia in the infarcted area on both ventriculography and MRI, with a reduced myocardial accumulation of 99mTc MIBI (66.3%+/-11.8% vs 95.6%+/-2.2% in group B). The mean wall thickness during the complete cardiac cycle (9.5+/-1.8 mm vs 13.1+/-1.1 mm in group B, P<0.001), the systolic wall thickening (2.6+/-1.4 mm vs 5.8+/-1.5 mm in group B, P<0.01) and the phosphocreatine/adenosine triphosphate ratio (1.12+/-0.22 vs 1.74+/-0.23 in group B, P<0.01) in the hypokinetic area were all significantly reduced. It is concluded that persisting hypokinetic myocardium after NQWI combined with reduced myocellular uptake of 99mTc-MIBI displays a reduced PCr/ATP ratio. Our results indicate that morphological remodelling after NQWI is accompanied by fundamental changes in cardiac energy metabolism. PMID- 11383866 TI - Determination of right ventricular ejection fraction from reprojected gated blood pool SPET: comparison with first-pass ventriculography. AB - Gated blood pool (GBP) studies are widely available and relatively inexpensive. We have previously published a simple and convenient method for measuring left ventricle ejection fraction (EF) with increased accuracy from single-photon emission tomography (SPET) GBP scans. This paper describes an extension of this method by which right ventricular EF may also be measured. Gated SPET images of the blood pool are acquired and re-oriented in short-axis slices. Counts from the left ventricle are excluded from the short-axis slices, which are then reprojected to give horizontal long-axis images. Time-activity curves are generated from each pixel around the right ventricle, and an image is created with non-ventricular pixels "greyed out". This image is used as a guide in drawing regions of interest around the right ventricle on the end-diastolic and end-systolic long-axis images. In 28 patients, first-pass ventriculography studies were acquired followed by SPET GBP scans. The first-pass images were analysed a total of four times by two observers and the SPET images were analysed three times each by two observers. The agreement between the two techniques was good, with a correlation coefficient of 0.72 and a mean absolute difference between first-pass and reprojected SPET EFs of 4.8 EF units. Only four of the 28 patients had a difference of greater than 8 EF units. Variability was also excellent for SPET right ventricular EF values. Intra-observer variability was significantly lower for SPET than for first-pass EFs: standard error of the estimate (SEE)=5.1 and 7.3 EF units, respectively (P<0.05). Inter-observer variability was comparable in the two techniques (SEE=5.2 and 6.9 EF units for SPET and first-pass ventriculography, respectively). PMID- 11383867 TI - Concordance between rest MIBG and exercise tetrofosmin defects: possible use of rest MIBG imaging as a marker of reversible ischaemia. AB - Perfusion imaging combined with pharmacological stress is the study of choice in patients with ischaemic heart disease who are incapable of exercising. Some medical conditions, however, can preclude the use of pharmacological stress. In these particular situations, availability of a diagnostic test which allows for the assessment of ischaemic territory at rest would be desirable. With the purpose of providing a marker of reversible ischaemia, we evaluated myocardial iodine-123 metaiodobenzylguanidine (MIBG) uptake in regions with fixed and reversible defects defined by exercise/rest perfusion study. Fifty-four male patients with ischaemic heart disease and previous myocardial infarction were studied by means of exercise/rest tetrofosmin and MIBG single-photon emission tomography (SPET). Regional tracer uptake was quantified and expressed as a percentage of maximum peak activity. Areas with denervated but perfused myocardium and areas with ischaemic myocardium were calculated. Regions with<75% of peak activity in the exercise perfusion study were divided into two groups according to whether the increase in peak activity in the respective rest study was >10% (reversible regional defect) or <10% (fixed regional defect). These percentages were compared with the percentages of the innervation study. The area of the innervation defect was significantly larger when the perfusion defect was reversible than when it was fixed. In regions with reversible perfusion defects, the size of the area of denervated but perfused myocardium was similar to the size of the area of ischaemic myocardium. In regions with reversible defects, the percentage of myocardial MIBG uptake was not significantly different from the percentage of tetrofosmin uptake at exercise, while it was significantly lower than the percentage of tetrofosmin uptake at rest. In regions with fixed defects, the percentage of myocardial MIBG uptake was significantly lower than the percentage of tetrofosmin uptake at exercise and at rest. In patients who developed angina during exercise test, the area of denervated but perfused myocardium was significantly larger than in patients without angina (4.1+/-2.4 vs 3.4+/-2.5, P=0.02). The same trend was observed with regard to the size of the innervation defect (8.6+/-2.4 vs 5.7+/-2.2, P=0.02). It is concluded that when the use of pharmacological stress is not possible in patients incapable of exercising, rest studies with MIBG combined with rest myocardial perfusion studies may be useful as a marker of reversible ischaemia. PMID- 11383868 TI - Age- and gender-specific differences in left ventricular cardiac function and volumes determined by gated SPET. AB - The aim of this study was to determine normative volumetric data and ejection fraction values derived from gated myocardial single-photon emission tomography (SPET) using the commercially available software algorithm QGS (quantitative gated SPET). From a prospective database of 876 consecutive patients who were referred for a 2-day stress-rest technetium-99m tetrofosmin (925 MBq) gated SPET study, 102 patients (43 men, 59 women) with a low (<10%) pre-test likelihood of coronary disease were included (mean age 57.6 years). For stress imaging, a bicycle protocol was used in 79 of the patients and a dipyridamole protocol in 23. Left ventricular ejection fraction (LVEF) and end-diastolic and -systolic volumes (EDV and ESV) were calculated by QGS. EDV and ESV were corrected for body surface area, indicated by EDVi and ESVi. To allow comparison with previous reports using other imaging modalities, men and women were divided into three age groups (<45 years, > or =45 years but <65 years and > or =65 years). Men showed significantly higher EDVi and ESVi values throughout and lower LVEF values when compared with women in the subgroup > or =65 years (P<0.05, ANOVA). Significant negative and positive correlations were found between age and EDVi and ESVi values for both women and men and between LVEF and age in women (Pearson P< or =0.01). LVEF values at bicycle stress were significantly higher than at rest (P=0.000, paired t test), which was the result of a significant decrease in ESV (P=0.003), a phenomenon which did not occur following dipyridamole stress (P=0.409). The data presented suggest that LVEF and EDVi and ESVi as assessed by QGS are strongly gender-specific. Although the physiological significance of these results is uncertain and needs further study, these findings demonstrate that the evaluation of cardiac function and volumes of patients by means of QGS should consider age- and gender-matched normative values. PMID- 11383869 TI - Temporal hypometabolism at the onset of cryptogenic temporal lobe epilepsy. AB - Most patients with intractable temporal lobe epilepsy (TLE) exhibit temporal glucose hypometabolism. The reasons for the development of this abnormality are as yet unclear. The current notion is that an initial injury causes seizures, which in turn give rise to hypometabolism. The aim of this study was to assess whether temporal reductions in glucose metabolism in non-lesional TLE are the result of repeated seizures or whether hypometabolism represents an initial disturbance at the onset of disease. Glucose consumption was assessed with fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG PET) in 62 patients with cryptogenic non-refractory TLE in different stages of disease. Twelve subjects without neurological illness served as controls. Patients with onset of epilepsy at least 3 years prior to the PET scan were defined as having chronic TLE. Using this criterion, the whole patient cohort included 27 patients with de novo TLE and 35 patients with chronic TLE. The groups were matched for age and sex. The appearance of high-resolution magnetic resonance images of the brain was unremarkable in all patients. In the total cohort, number, duration and frequency of seizures had a significant relation to the magnitude of hypometabolism. Temporal hypometabolism was exhibited by 26 of the 62 patients (42%), including 8 out of 27 (30%) with newly diagnosed TLE and 18 out of 35 (51%) with chronic TLE. The disturbances were more extensive and more severe in patients with chronic TLE. It is concluded that temporal hypometabolism may already be present at the onset of TLE, but is less frequent and less severe in newly diagnosed than in chronic TLE. The metabolic disturbance correlates with the number of seizures. These findings suggest that an initial dysfunction is present in a considerable number of patients and that hypometabolism is worsened by continuing epileptic activity. PMID- 11383870 TI - Iodide uptake in human anaplastic thyroid carcinoma cells after transfer of the human thyroid peroxidase gene. AB - Human thyroperoxidase (hTPO) is critical for the accumulation of iodide in thyroid tissues. Poorly differentiated and anaplastic thyroid tumours which lack thyroid-specific gene expression fail to accumulate iodide and, therefore, do not respond to iodine-131 therapy. We consequently investigated whether transfer of the hTPO gene is sufficient to restore the iodide-trapping capacity in undifferentiated thyroid and non-thyroid tumour cells. The human anaplastic thyroid carcinoma cell lines C643 and SW1736, the rat Morris hepatoma cell line MH3924A and the rat papillary thyroid carcinoma cell line L2 were used as in vitro model systems. Employing a bicistronic retroviral vector based on the myeloproliferative sarcoma virus for the transfer of the hTPO and the neomycin resistance gene, the C643 cells and SW1736 cells were transfected while the L2 cells and MH3924A cells were infected with retroviral particles. Seven recombinant C643 and seven SW1736 cell lines as well as four recombinant L2 and four MH3924A cell lines were established by neomycin selection. They were studied for hTPO expression using an antibody-based luminescence kit, followed by determination of the enzyme activity in the guaiacol assay and of the iodide uptake capacity in the presence of Na125I. Genetically modified cell lines expressed up to 1,800 times more hTPO as compared to wild type tumour cells. The level of hTPO expression varied significantly between individual neomycin resistant cell lines, suggesting that the recombinant retroviral DNA was integrated at different sites of the cellular genome. The accumulation of iodide, however, was not significantly enhanced in individual recombinant cell lines, irrespective of low or high hTPO expression. Moreover, there was no correlation between hTPO expression and enzyme activity in individual cell lines. The transduction of the hTPO gene per se is not sufficient to restore iodide trapping in non-iodide-concentrating tumour cells. Future studies will have to concentrate on the possible expression of enzymatically active proteins or the transfer of multiple genes involved in iodide trapping. PMID- 11383872 TI - Summary of the Second International Sentinel Node Conference. AB - A personal account is given of the most notable developments in lymphatic mapping that were discussed at the international conference "Sentinel Node 2000", held in December 2000 in Santa Monica. The latest advances in tracers, imaging, instrumentation and radiation are first outlined. Thereafter, techniques, results and controversies with respect to melanoma, breast cancer and other cancers are discussed. It is concluded that lymphatic mapping is quickly gaining momentum. The technique that is best used for lymphatic mapping is well established in melanoma but not in breast cancer. The results of large randomised studies are awaited in both these diseases. Gastrointestinal cancer is the main new focus. PMID- 11383871 TI - Relationship between expression of the sodium/iodide symporter and 131I uptake in recurrent lesions of differentiated thyroid carcinoma. AB - The sodium/iodide symporter (NIS) is known to be responsible for the active accumulation of iodide within the thyroid gland. We evaluated the relationship between the expression of NIS in primary or lymph node lesions and iodine-131 uptake in recurrent lesions of differentiated thyroid cancer. In 67 patients with differentiated thyroid cancer (5 follicular and 62 papillary carcinomas), the expression of NIS was analysed by immunohistochemical staining using polyclonal antibodies against human NIS. We used paraffin block tissues of primary tumours or metastatic lesions, and also assessed 131I uptake in recurrent lesions of thyroid cancer on post-operative 131I whole-body scan. Immunohistochemical staining was positive in 22 patients (32.8%), including 2 of 5 follicular and 20 of 62 papillary carcinomas. Recurrence was confirmed in 40 patients pathologically or clinically by serum thyroglobulin, 131I scan, fluorine-18 fluorodeoxyglucose positron emission tomography and/or computed tomography. Among these 40 patients, 28 showed positive uptake on 131I scan. Fourteen tumour specimens out of 28 (50%) were positive by NIS immunohistochemical staining. The remaining 12 patients with recurrent cancer showed negative 131I scans, and all specimens were negative by NIS immunohistochemical staining. Thus, NIS immunohistochemical staining predicted 131I uptake in recurrent cancer with a 100% positive predictive value and a 46.2% negative predictive value. There was no difference in the positivity of NIS according to the site of recurrence on 131I scan. Outcome of 131I therapy could be assessed in 22 of the 28 patients who showed 131I uptake in recurrent lesions. Patients with positive NIS immunostaining responded to 131I therapy better than did patients with negative immunostaining (P<0.05). In conclusion, NIS immunohistochemical staining showed a high positive predictive value in predicting iodine uptake. Positive immunohistochemical staining of human NIS in primary or lymph node lesions may predict 131I accumulation and effectiveness of 131I therapy in recurrent lesions. PMID- 11383873 TI - Carbon-11 acetate as a tracer of myocardial oxygen consumption. AB - Estimation of myocardial oxygen consumption (MVO2) and myocardial blood flow (MBF) is important for the understanding of various (patho)physiological mechanisms and diseases. Clearance rates of carbon-11 labelled acetate, determined with positron emission tomography, allow estimation of MVO2 on a segmental level and non-invasively. In addition, MBF can be determined from uptake rates. In this review, the background to estimation of MVO2 and MBF is discussed, as well as the currently available literature that has used 11C acetate to estimate MVO2 and MBF. PMID- 11383874 TI - 131I thyroid uptake in patients treated with 131I-Lipiodol for hepatocellular carcinoma. PMID- 11383875 TI - Morphological analysis of CD15-immunoreactive neurons in the guinea pig retina. AB - Using immunocytochemistry, morphometry and electron microscopy, we have investigated the distribution and characteristics of CD15-immunoreactive (IR) neurons in the guinea pig retina. In the present study, two types of amacrine cells, including interplexiform cells in the inner nuclear layer (INL) and some cells in the ganglion cell layer (GCL), were labeled with anti-CD15 antisera. Type 1 amacrine cells had large somata located in the INL, with long and branched processes ramifying mainly in strata 4 and 5 of the inner plexiform layer (IPL). Somata of type 2 cells had smaller diameters, and were also located in the INL. Their processes stratified in stratum 1. The densities of type I and type 2 amacrine cells increased from 152.8+/-36.7/mm2 and 160.6+/-61.7/mm2 in the peripheral retina, to 404.3+/-41.5/mm2 and 552.2+/-72.2/mm2 in the central retina, respectively. Cells in the GCL exhibiting CD15 immunoreactivity were rarely observed. Colocalization experiments, using consecutive semi-thin sections, demonstrated that these CD15-IR amacrine cells exhibited gamma aminobutyric acid (GABA) immunoreactivity. In addition, the processes of the type 1 cells formed one member of the postsynaptic dyads that are formed in the axon terminals of rod bipolar cells. Most of these processes made reciprocal synapses back to the axon terminals of the rod bipolar cells. Thus, CD15-IR amacrine cells constitute a subpopulation of GABAergic amacrine cells in the guinea pig retina, and the type 1 cells among them provide the inhibitory input to rod bipolar cells. PMID- 11383876 TI - Lipopolysaccharide alters ecto-ATP-diphosphohydrolase and causes relocation of its reaction product in experimental intrahepatic cholestasis. AB - Lipopolysaccharide (LPS), a bacterial endotoxin, exerts profound inflammatory actions toward various tissues and cells. We induced intrahepatic cholestasis in rats by administration of LPS and followed ecto-ATP-diphosphohydrolase (ecto apyrase) activity in the liver. The activity of the enzyme had decreased to 77% 2 h after injection compared with the activity in control animals. The maximum decrease was detected 24 h after administration. The activity was found to have partially recovered 1 week after injection, but had yet to reach control levels. In contrast to the decrease in ecto-apyrase activity, there were increases in alkaline phosphatase activity and bilirubin concentration, markers of cholestasis. In response to LPS, the reaction product of ecto-apyrase was found to relocate from the canalicular domain of the plasma membrane of hepatocytes, its predominant localization in the liver of intact animals, to the basolateral and sinusoidal domains. The pattern of histochemical reaction indicated modulation of the enzyme activity and changes in trafficking of intracellular proteins. Taken together, our findings showed that LPS administration alters ecto apyrase and causes relocation of its reaction product from the canalicular domain of the plasma membrane of hepatocytes in the rat. It is suggested that relocation of the reaction product may be a protective mechanism to enable the hepatocytes to withstand the cytokine-induced metabolic perturbations. PMID- 11383878 TI - Effect of different photoperiods on the ultrastructure of the specific secretory cells and alpha-subunit mRNA level in the chicken pars tuberalis. AB - The effect of different photoperiods on the specific secretory cells of the pars tuberalis was examined in male chicks. Animals were placed in one of three different photoperiod regimens: (1) normal control (light:dark = 12 h:12 h), (2) continuous light (L:D = 24 h:0), and (3) extended darkness (L:D = 1 h:23 h). The levels of common alpha-subunit mRNA in the pars tuberalis were examined by Northern blot analysis and compared with those in the pars distalis. In chicks exposed to continuous light for 1 week, alpha-subunit mRNA level in the pars tuberalis was decreased, although the level in the pars distalis was increased. Exposure to continuous light for 30 days also induced a decrease in alpha-subunit mRNA level in the pars tuberalis. On the other hand, in chicks exposed to extended darkness for 1 week, the alpha-subunit mRNA level of the pars tuberalis was markedly increased. In situ hybridization with digoxigenin-labeled common alpha-subunit cRNA probe also showed that the hybridization signals for alpha subunit mRNA in the pars tuberalis cells become weak under continuous light for 30 days but they are very intense under extended darkness. Thus, the synthesis of alpha-subunits in the chick pars tuberalis was inhibited by continuous light but stimulated by extended darkness. These results were confirmed by semiquantitative electron-microscopic analyses. After exposure to continuous light for 30 days, many pars tuberalis (PT)-specific cells were filled with enlarged secretory granules, showing the reduction of secretory activity. On the contrary, extended darkness for 30 days induced hypertrophy of the PT-specific cells; the areas of cytoplasm and nucleus were significantly increased. In addition, secretory granules became small in size and exocytotic features were more frequent. Mitochondria and lysosomes were also increased in number. Thus, the synthetic and secretory activities of the PT-specific cells were increased under extended darkness. The data indicate that the specific cells of the pars tuberalis are responsive to photoperiodic changes in the chick. PMID- 11383877 TI - Reactive astrocytes upregulate one or more gene products that are recognized by monoclonal antibody H. AB - We generated a panel of monoclonal antibodies (mAbs) selected to recognize components from a Triton X-100 extract of ovine oligodendrocytes. One of these Abs, mAb H, recognizes an O-linked N-acetyl glucosamine residue in a specific conformation and/or environment. mAb H stained, weakly, two bands with Mr x 10( 3) of 209 and 62 in lysates of cultured rat astrocytes, suggesting antigens of low abundance. We have employed immunohistochemistry to investigate the cell and tissue distribution of the mAb H antigen(s). In normal rat and human brains, the sparse reaction products detected were confined, mostly, to fibrous astrocytes. In sharp contrast, when pathological specimens from a variety of brain lesions, including anisomorphic and isomorphic gliosis, were examined, a strong reaction with mAb H was in evidence in all reactive astrocytes, independent of the origin or nature of the lesions. This we interpret as meaning that the gene product(s) recognized by this mAb is (are) upregulated or induced following injury to the brain. Hence, epitope H represents a new addition to the list of molecules that are affected by brain injury. Structural and functional identification of the antigen(s) should shed light on its (their) relevance to the pathophysiology of the disease process. PMID- 11383879 TI - Intrasexual and intersexual dimorphisms of the red salmon prosencephalon. AB - Intrasexual as well as intersexual dimorphisms were found in the prosencephalon and mesencephalon of adult Oncorhynchus nerka (red/sockeye salmon). These dimorphisms are concerned with the position of the preoptic nucleus, nucleus lateralis tuberis, habenula, third ventricle, tectal ventricles, preoptic recess, recessus lateralis, horizontal commissure, posterior commissure, and toral commissure. The intrasexual dimorphism was characterized by either a rostral ("r" pattern) or a caudal ("c"-pattern) position of the preoptic region as well as varying locations of other structures within the prosencephalon. As compared to "c"-pattern fish, the preoptic nucleus and nucleus lateralis tuberis were located more rostral, and the habenula was positioned further caudal, in "r"-type animals. The intersexual dimorphism was also characterized by different positions of the structures listed above. With the exception of the preoptic nucleus, all of these were located further rostral in "r"-pattern females than in type "r" males. In "c"-pattern females, they were positioned further caudal than in type "c" males. The number of neurons in the parvocellular and in the magnocellular portion of the preoptic region differed in the two genders with respect to "r"- as well as "c"-pattern fish. Males had more neurons than females in both the magno- and the parvocellular subdivisions of the preoptic region. In "r"- and "c" pattern fish, the average size of magnocellular preoptic neurons was larger in females than in males. The observed intersexual variations may reflect gender specific differences in the control of the pituitary. Functional correlates of intrasexual dimorphism are obscure. PMID- 11383880 TI - Cell surface events during resealing visualized by scanning-electron microscopy. AB - The function of exocytosis during plasma membrane resealing might be to facilitate the flow of surface lipid over the disruption site and/or to add defect-spanning "patches" of internal membrane across it. Scanning-electron microscopic visualization of large plasma membrane disruptions in sea urchin eggs is here used to distinguish between these two possibilities. Disruptions were induced by shear stress in the presence and absence of resealing-permissive levels of external Ca2+, and the eggs were fixed at various intervals thereafter for microscopic processing. In eggs fixed immediately (<1 s) after shearing in the absence of Ca2+, a condition which prevents resealing, disruption sites were filled with a uniform population of spherical vesicles (approximately 1 microm in diameter). In eggs fixed immediately after shearing at a resealing-permissive level of Ca2+, disruption sites were filled with a highly heterogeneous population of enlarged vesicles, some being more than 10 microm in diameter and many having irregular profiles and/or appearing to be joined to one another. In eggs fixed 2 s or 5 s post-shearing, the continuity of these large vesicles with one another and the surface membrane began to obscure individual vesicle identities. Single "apertures" of discontinuity over disruption sites, the predicted morphology of a flow-based resealing mechanism, were not observed at any time point (1-5 s) during the interval required for completion of resealing. These observations provide strong confirmation that "patching" of large disruptions mediates their resealing. PMID- 11383881 TI - Quantitative analysis of epithelial cell aggregation in the simple metazoan Hydra reveals a switch from homotypic to heterotypic cell interactions. AB - Hydra, a member of the diploblastic phylum Cnidaria, exhibits the most basic type of organized metazoan tissues. Two unicellular sheets of polarized epithelial cells - ectoderm and endoderm - form a double layer throughout the body column. The double layer can be reestablished from single-cell suspensions by tissue specific cell-sorting processes. However, the underlying pattern of interactions between ectodermal and endodermal epithelial cells responsible for double-layer formation is unclear. By analyzing cell interactions in a quantitative adhesion assay using mechanically dissociated Hydra epithelial cells, we show that aggregation proceeds in two steps. First, homotypic interactions within ectodermal epithelial cells (ecto-ecto) and within endodermal epithelial cells (endo-endo) form homotypic cell clusters. Second, at an aggregate size of about ten epithelial cells/cluster, ectodermal and endodermal clusters start to form heterotypic aggregates. Homotypic ecto-ecto interactions are inhibited by a polyclonal anti-Hydra membrane antiserum, and under these conditions homotypic endo-endo interactions do not proceed beyond a size of about ten epithelial cells/cluster. These data suggest that homotypic cell clusters reduce their initial homotypic affinity and acquire a new heterotypic affinity. A link between cell adhesion and cell signaling in early Hydra aggregates is discussed. PMID- 11383882 TI - Morphological and functional characterization of beige mouse adrenomedullary secretory vesicles. AB - We tested whether the giant secretory granules observed in the mast cells of the naturally occurring mutant beige mouse (BM) (C57BL/6N-bg) were also present in the adrenal chromaffin cells. The presence of large chromaffin granules (CG) would be a valuable tool for the study of exocytosis in neuronal tissues. Conversely, the observation of large vesicles within chromaffin cells that are different from CG could indicate that CG are of a different origin than granules of mast cells. Ultrastructural analysis demonstrated the presence of large lysososmal-like vesicles in the BM, and also a discrete increase in the number of CG with diameters larger than 240 nm but not of giant CG. In addition, amperometric measurements of single-event exocytosis, using carbon fiber microelectrodes, showed no differences between the quantal size of secretory events from BM and wildtype or bovine chromaffin cells. Minor but significant differences were found between the kinetics of exocytosis in BM cells andwild type mouse cells. We conclude that CG, but not the abnormal-sized vesicles found in BM chromaffin cells contribute to the catecholamine secretion and that abnormal secretory granules are not present in adrenergic cell lineage. PMID- 11383883 TI - Immunoelectron-microscopic study of Kit-expressing cells in the jejunum of wildtype and Ws/Ws rats. AB - Interstitial cells of Cajal (ICC) are responsible for generating electrical slow waves in the gastrointestinal (GI) tract. Slow waves regulate the frequency of contractions of the tunica muscularis, and therefore ICC are critical for normal motility in the small intestine. ICC express Kit, the gene product of c-kit, a protooncogene that encodes a receptor tyrosine kinase. Physiological evidence demonstrating that ICC are pacemakers has come from experiments on W-mutant mice which have few Kit-positive cells at the level of the myenteric plexus (IC-MY) and also lack electrical slow waves. In the past identification of ICC required the use of electron microscopy, however the discovery that ICC express Kit has facilitated studies of the distribution of ICC in several species. Immunoelectron microscopy to relate ultrastructure to Kit expression has only been performed in a limited number of studies of mice. We examined the ultrastructure of Kit expressing cells in the rat using immunoelectron microscopy and an anti-Kit antibody. We compared the presence and appearance of Kit-expressing ICC in wildtype and Ws/Ws rats, which carry a mutation in the white spotting locus and have a phenotype similar to W/Wv mutant mice. Kit-expressing cells could be detected in the myenteric plexus (MY) and deep muscular plexus (DMP) regions of the small intestine of wildtype animals. In Ws/Ws rats, Kit-expressing cells were not observed in the region of MY, but were observed in the DMP. The density of Kit-positive cells in the DMP of Ws/Ws rats was similar to those in wildtype rats. Electron microscopy showed that Kit-expressing cells at the level of the MY of the rat had similar ultrastructural features as IC-MY in wildtype mice. IC-DMP in the rat of both wildtype and Ws/Ws mutants were similar in structure to IC-DMP of the mouse. We conclude that wildtype rats have IC-MY and IC-DMP in the tunica muscularis of the jejunum. ICC express Kit-like immunoreactivity (Kit-LI) in the rat as in the mouse. IC-MY are absent in the small intestine of Ws/Ws rats, and this corresponds to the lack of Kit-labeling in this region. Ws/Ws rats, however, possess IC-DMP with normal ultrastructural features and Kit-LI. The absence of IC MY of Ws/Ws rats is likely to account for the abnormal contractile activity of the GI tract observed in these mutants. The present study suggests that Ws/Ws rats could provide an interesting model to investigate the physiological significance of pacemaker activity because they manifest a defect in IC-MY. PMID- 11383884 TI - Distribution of nitric oxide synthase and vasoactive intestinal polypeptide immunoreactivity in the sphincter of Oddi and duodenum of the possum. AB - The nitrergic innervation of the sphincter of Oddi (SO) and duodenum in the Australian brush-tailed possum and the possible association of this innervation with the neuropeptide vasoactive intestinal polypeptide (VIP) were investigated by using immunohistochemical localisation of nitric oxide synthase (NOS) and VIP, together with the general neuronal marker, protein gene product 9.5 (PGP9.5). Whole-mount preparations of the duodenum and attached SO without the mucosa, submucosa and circular muscle (n=12) were double- and triple-labelled. The density of myenteric nerve cell bodies of the SO in the more distal region (duodenal end) was significantly higher than that in the more proximal region. In the SO, approximately 50% of all cells were NOS-immunoreactive (IR), with 27% of the NOS-IR cells being VIP-IR. Within the duodenal myenteric plexus, NOS immunoreactivity was present in about 25% of all neurons, with 27% of these NOS IR neurons also being VIP-IR, a similar proportion to that in the SO. Varicose nerve fibres with NOS and VIP immunoreactivity were present within the myenteric and submucous plexuses of the SO and duodenum, and in the circular and longitudinal muscle layers. The NOS-positive cells within both the SO and duodenum were unipolar, displaying a typical Dogiel type I morphology. The myenteric plexuses of the SO and duodenum were in direct continuity, with many interconnecting nerve trunks, some of which showed NOS and VIP immunoreactivity. Thus, the possum possesses an extensive NOS innervation of the SO and duodenum, with a significantly higher proportion of NOS-IR neurons within the SO, a subset of which contains VIP. PMID- 11383885 TI - Differential localization of prohormone convertases PC1 and PC2 in two distinct types of secretory granules in rat pituitary gonadotrophs. AB - Prohormone convertases PC1 and PC2 are endoproteases involved in prohormone cleavage at pairs of basic amino acids. There is a report that prohormone convertase exists in the rat anterior pituitary gonadotrophs, where it had previously been considered that proprotein processing does not take place. In addition to luteinizing hormone and follicle-stimulating hormone, rat pituitary gonadotrophs contain chromogranin A (CgA) and secretogranin II (SgII), two members of the family of granin proteins, which have proteolytic sites in their molecules. In the present study we examined whether there is a close correlation between subcellular localization of prohormone convertases and granin proteins. Ultrathin sections of rat anterior pituitary were immunolabeled with anti-PC1 or PC2 antisera and then stained with immunogold. Immunogold particles for PC1 were exclusively found in large, lucent secretory granules, whereas those for PC2 were seen in both large, lucent and small, dense granules. The double-immunolabeling also demonstrated colocalization of PC2 and SgII in small, dense granules and of PC1, PC2, and CgA in large, lucent granules. These immunocytochemical results suggest that PC2 may be involved in the proteolytic processing of SgII and that both PC1 and PC2 may be necessary to process CgA. PMID- 11383886 TI - Effect of protracted estrogen administration on the thyroid of Ames dwarf mice. AB - The effect of protracted estrogen administration on estrogen receptor expression and cellular composition of the thyroid was examined in genetically thyrotropin (TSH)-deficient female Ames dwarf mice (df/df) to reveal whether estrogen might act independently from TSH. inducing changes in thyroid morphology and function. To evaluate such changes, the thyroid from four estrogen-implanted Ames dwarf mice, four sham-implanted Ames dwarf mice and four sham-implanted normal littermate mice were investigated histologically, immunohistochemically and morphometrically. Our morphologic study demonstrated significant differences in the colloid areas of normal and dwarf mice (P<0.001). The correlation observed between this parameter and body weights (r=0.610, P<0.05) and thyroid weights (r=0.729, P<0.01) suggests that the decrease in the colloid areas is not a result of abnormal folliculogenesis but is in direct correlation with the small thyroid and body size of dwarf mice. Although two types of estrogen receptors are known to exist in the present study, only the alpha (ERalpha) variant was found in the thyroid. ERalpha immunoreactivity was detected in the nuclei of parafollicular cells but not of the follicular epithelium. No significant differences were reported in ER expression between estrogen-implanted dwarf mice and sham implanted dwarf mice, suggesting that estrogen receptor expression in the thyroid is independent of circulating estrogen levels. In spite of the absence of ERalpha in follicular cells, protracted estrogen administration affected mainly the follicular cells. Our results suggest that when TSH is absent estrogens may exert a negative feedback on the activity of follicular cells. PMID- 11383887 TI - Asymmetrical development of bones and soft tissues during eye migration of metamorphosing Japanese flounder, Paralichthys olivaceus. AB - The symmetrical body of flatfish larvae dramatically changes into an asymmetrical form after metamorphosis. Eye migration results in the most significant asymmetrical development seen in any vertebrate. To understand the mechanisms involved in eye migration, bone and cartilage formation was observed during metamorphosis in laboratory-reared Japanese flounder, Paralichthys olivaceus, by using whole-body samples and histological sections. Most of the hard tissues of the cranium (parasphenoid, trabecular cartilage, supraorbital canal, and supraorbital bar) exist symmetrically in the larval period before metamorphosis and develop by twisting in the same direction as that in which the eye migrates. An increase in skin thickness beneath the eye was observed only on the blind side at the beginning of eye migration; this was the first definitive difference between the right and left sides of the body. The pseudomesial bar, a peculiar bone present only in flatfishes, developed from this thick skin and grew dorsad. Novel sac-like structures were found and named retrorbital vesicles. The retrorbital vesicle of the blind side grew larger and faster than that of the ocular side when the right eye moved most dramatically, whereas no difference was observed between the volume of right and left connective tissue in the head. The asymmetrical presence and growth of the pseudomesial bar together with inflation of the retrorbital vesicle on the blind side may be responsible for right eye migration during metamorphosis in the Japanese flounder. PMID- 11383888 TI - Formation of the sphenomandibular ligament by Meckel's cartilage in the mouse: possible involvement of epidermal growth factor as revealed by studies in vivo and in vitro. AB - In mammals, the midportion of the soft tissue of Meckel's cartilage at the degenerating stage forms a ligament known as the sphenomandibular ligament. To clarify the mechanism of formation of this ligament by Meckel's cartilage in mouse, we examined the effects of epidermal growth factor (EGF) on the chondrocytes in terms of the proliferation and differentiation of cells and calcification of the matrix in vivo and in vitro. The effects of EGF were examined by immunohistochemical staining, with EGF-soaked beads, by electron microscopy, and by general histochemical analysis of proteoglycans and calcification. Analysis of labeling with bromodeoxyuridine (BrdU) and the rate of cell growth revealed that EGF enhanced DNA synthesis and the proliferation of Meckel's chondrocytes. Histological findings in organ culture and in cell culture, with and without the application of EGF-soaked beads, revealed that EGF inhibited the differentiation of cells to chondrocytes and induced phenotypic changes in fibroblastic cells. The inhibition of alkaline phosphatase activity that resulted from exposure to EGF was accompanied by prolonged calcification of the matrix. Whole-mount staining revealed that subcutaneous injection of EGF enhanced the disappearance of Meckel's cartilage. Our results suggest a possible mechanism whereby the midportion of Meckel's cartilage remains uncalcified and is rapidly transformed into the sphenomandibular ligament. PMID- 11383889 TI - Sexual differences and effects of castration on secretory mode and intracellular calcium ion dynamics of golden hamster Harderian gland. AB - The aim of the present work was to study the sexual differences in secretory mechanisms and intracellular calcium ion dynamics in the Harderian gland of the golden hamster. In both sexes the Harderian gland consisted of small and large lobes. In the intact control male glands the secretory portions of both lobes showed wide lumina that contained secretory material and cytoplasmic fragments, suggestive of the occurrence of exocytosis and apocrine secretion. After perfusion with HEPES-buffered Ringer's solution containing 10 microM carbamylcholine (CCh), the glandular cells showed features of enhanced secretion and a rise in intracellular calcium concentration ([Ca2+]i). In the intact control female gland the lumina of most secretory portions in the large lobe contained porphyrin accretions, and exocytosis was the sole secretory mechanism. Stimulation of the large lobe with 10 microM CCh did not raise [Ca2+]i or cause enhanced secretion. The small lobe in females resembled the male gland in secretory functions, and CCh administration caused enhanced secretion and a rise in [Ca2+]i. Castration in males abolished apocrine secretion; exocytosis became the sole secretory mechanism, and stimulation of the glandular cells with CCh did not cause enhanced secretion or induce a rise in [Ca2+]i. To the contrary, in females, castration restored apocrine secretion and CCh administration caused enhanced secretion and a rise in [Ca2+]i. Castration did not affect the secretory mechanisms and the effect of CCh on the glandular cells in the small lobes of both male and female glands. The present study points to the possibility that sex hormones may control the functioning or expression of muscarinic receptors in the Harderian gland of the golden hamster. PMID- 11383890 TI - The distribution of MUC1, an apical membrane glycoprotein, in mammary epithelial cells at the resolution of the electron microscope: implications for the mechanism of milk secretion. AB - The distribution of the glycoprotein, mucin 1 (MUC1), was determined in lactating guinea-pig mammary tissue at the resolution of the electron microscope. MUC1 was detected on the apical plasma membrane of secretory epithelial cells, the surface of secreted milk-fat globules, the limiting membranes of secretory vesicles containing casein micelles and in small vesicles and tubules in the apical cytoplasm. Some of the small MUC1-containing vesicles were associated with the surfaces of secretory vesicles and fat droplets in the cytoplasm. MUC1 was detected in much lower amounts on basal and lateral plasma membranes. By quantitative immunocytochemistry, the ratio of MUC1 on apical membranes and milk fat globules to that on secretory vesicle membranes was estimated to be 9.2:1 (density of colloidal gold particles/microm membrane length). The ratio of MUC1 on apical membranes compared with basal/lateral membranes was approximately 99:1. The data are consistent with a mechanism for milk-fat secretion in which lipid globules acquire an envelope of membrane from the apical surface and possibly from small vesicles containing MUC1 in the cytoplasm. During established lactation, secretory vesicle membrane does not appear to contribute substantially to the milk-fat globule membrane, or to give rise in toto to the apical plasma membrane. PMID- 11383891 TI - A new synthetic method for preparing indole derivatives from 2-keto glycosides. AB - In this article, a new reaction of the addition of two molecules of aniline to 2 keto glycosides (glycoside 2-uloses, 2-ulosides) is reported. A possible pathway for the reaction is presented. This reaction provides a novel one-pot method for the synthesis of indole derivatives from sugars. PMID- 11383892 TI - Synthesis and identification in bacterial lipopolysaccharides of 5,7-diacetamido 3,5,7,9-tetradeoxy-D-glycero-D-galacto- and -D-glycero-D-talo-non-2-ulosonic acids. AB - 5,7-Diacetamido-3,5,7,9-tetradeoxy-D-glycero-D-galacto- and -D-glycero-D-talo-non 2-ulosonic acids were synthesized by condensation of 2,4-diacetamido-2,4,6 trideoxy-D-mannose with oxalacetic acid. Comparison of the 1H and 13C NMR data and the specific optical rotation values of these monosaccharides and the corresponding L-glycero-D-galacto and L-glycero-D-talo isomers synthesized earlier [Tsvetkov, Y. E.; Shashkov, A. S.; Knirel, Y. A.; Backinowsky, L. V.; Zahringer, U. Mendeleev Commun. 2000, 90-92] with data of the natural compounds enabled the identification in bacterial lipopolysaccharides of derivatives of 5,7 diamino-3,5,7,9-tetradeoxy-D-glycero-D-galacto-non-2-ulosonic (legionaminic) acid and epimers of legionaminic acid at C-4 and C-8. PMID- 11383893 TI - New evidence for the mechanism of the tin(II) chloride catalyzed reactions of vicinal diols with diazodiphenylmethane in 1,2-dimethoxyethane. AB - A kinetic study of the tin(II) chloride catalyzed reaction of diazodiphenylmethane with ethylene glycol in dimethoxyethane is reported. The preparation and characterization of ethylene glycol monodiphenylmethyl ether, the main product from this reaction, is also reported as well as the preparation of the two diphenylmethyl monoethers of methyl 4,6-O-benzylidene-alpha-D glucopyranoside. An unexpected relationship between the concentration of ethylene glycol and the pseudo first-order rate constant, k', was observed in these reactions. For low concentrations of ethylene glycol (below 0.06 M), k' increases with increasing concentration of the diol. This trend is reversed for high concentrations of ethylene glycol (from about 0.06 to about 0.2 M). The apparent rate constant was also inversely related to the initial concentration of diazodiphenylmethane for the concentrations investigated. These results make the previously proposed involvement of a 1,3,2-dioxastannolane intermediate very unlikely [Petursson, S.; Webber, J.M. Carbohydr. Res. 1982, 103, 41-52]. The results suggest that more likely intermediates for these reactions involve tin(II) chloride complexes in a dynamic equilibrium with the diol. PMID- 11383894 TI - Synthesis of methyl alpha-D-glucopyranosyl-(1-->4)-alpha-D-galactopyranoside and methyl alpha-D-xylo-hex-4-ulopyranosyl-(1-->4)-alpha-D-galactopyranoside. AB - The syntheses of methyl alpha-D-glucopyranosyl-(1-->4)-alpha-D-galactopyranoside (1) and methyl alpha-D-xylo-hex-4-ulopyranosyl-(1-->4)-alpha-D-galactopyranoside (4) are reported. The keto-disaccharide 4 is of interest in our design, synthesis, and study of pectate lyase inhibitors. The key step in the syntheses was the high-yielding, stereospecific formation of methyl 4,6-O-benzylidene-2',3' di-O-benzyl-alpha-D-glucopyranosyl-(1-->4)-2,3,6-tri-O-benzyl-alpha-D galactopyranoside (15), which was accomplished by reacting 2,3-di-O-benzyl-4,6-O benzylidene-D-glucopyranosyl trichloroacetimidate (10) with methyl 2,3,6-tri-O benzyl-alpha-D-galactopyranoside (14) in the presence of a catalytic amount of tert-butyldimethylsilyl trifluoromethane sulfonate (TMSOTF). Compound 15 was either hydrogenolyzed to yield disaccharide 1 or treated with NaBH3CN-HCl in 1:1 tetrahydrofuran-ether to yield methyl 2,3,6-tri-O-benzyl-alpha-D-glucopyranosyl (1-->4)-2,3,6-tri-O-benzyl-alpha-D-galactopyranoside (2). The free 4'-OH of compound 2 was oxidized to a carbonyl group by a Swern oxidation, and the protecting groups were removed by hydrogenolysis to yield keto-disaccharide 4. These synthetic pathways were simple, yet high yielding. PMID- 11383895 TI - A study of the donor properties of sialyl phosphites having an auxiliary 3-(S) phenylseleno group. AB - Two phosphite sialyl donors, each having an auxiliary 3-(S)-phenylseleno group, were prepared and evaluated. The phenylseleno group was introduced via a new mode of generating phenylselenenic acid ('PhSeOH'). Although the sialyl donors provided fair yields (32-76%) of the desired sialosides in glycosylations of the reactive acceptor 1,2;3,4-di-O-isopropylidene-alpha-D-galactopyranose, no sialylated products could be obtained with less reactive acceptors. The presence of a 5-N-acetylacetamido group on the phosphite sialyl donor did not appear to improve its sialylating capability. The weak C-Se bond, possibly in combination with a steric hindrance, which disfavors alpha-nitrilium ion formation, seem to explain the unsuccessful sialylations of the less reactive acceptors. PMID- 11383896 TI - Location of O-acetyl groups in S-657 using the reductive-cleavage method. AB - A two-step procedure is described in this paper to identify the position of O acetyl groups in S-657 polysaccharide. Reductive-cleavage experiments performed on the fully methylated (base-catalyzed) polysaccharide, followed by acetylation of anhydroalditols, identified individual sugar residues and their position of linkage. In a second experiment, the polysaccharide was methylated under neutral conditions leaving native acetate groups intact. Reductive cleavage of the neutral methylated polysaccharide using CF3SO3SiMe3 as a catalyst, followed by acetylation in situ, identified sugar residues containing native acetate groups and established their position of substitution. Using this two-step procedure of analysis, S-657 polysaccharide is shown to contain O-acetyl groups on the 2 position and the 2,6-positions of 3-linked glucopyranosyl residues. PMID- 11383897 TI - On the structure of kappa/iota-hybrid carrageenans. AB - The coil-to-helix transition and temperature dependence of the viscosity of commercial kappa/iota-hybrid carrageenans produced by the red algae Sarcothalia crispata, Mazaella laminarioides, and Chondrus crispus were studied using rheometry and optical rotation. The structure of these kappa/iota-hybrid carrageenans was determined by 1H and 13C NMR spectroscopy combined with monosaccharide composition analysis. The coil-to-helix transitions, measured by polarimetry and rheometry, of the kappa/iota-hybrid carrageenans are significantly different from those of pure kappa- and iota-carrageenan, and from hand-made mixtures thereof. This provides evidence that the kappa/iota-hybrid carrageenans are mixed chains, containing both kappa- and iota-repeating units. PMID- 11383898 TI - Structural studies of S-7, another exocellular polysaccharide containing 2-deoxy arabino-hexuronic acid. AB - The exocellular polysaccharide S-7, a heteropolysaccharide from Azotobacter indicus var. myxogenes has been studied using methylation analysis, Smith degradation, partial acid hydrolysis, NMR spectroscopy and mass spectrometry as the principal methods. It is concluded that the repeating unit has the following structure: [structure: see text] The absolute configuration of the deoxyhexuronic acid was deduced from 1H NMR chemical shifts and is most likely D. Approximately two O-acetyl groups per repeating unit are present, one of which is presumably on the Rha residue. The structure bears great resemblance to another polysaccharide, recently studied, produced by Sphingomonas paucimobilis I-886. PMID- 11383899 TI - Purification and characterisation of a galactoglucomannan from kiwifruit (Actinidia deliciosa). AB - A galactoglucomannan (GGM) has been purified from the primary cell walls of ripe kiwifruit. A combination of barium hydroxide precipitation, anion exchange- and gel-permeation chromatography gave a chemically homogeneous polymer with a 1:2:2 galactose-glucose-mannose ratio and a molecular weight range of 16-42 kDa. Complete hydrolysis of the polymer with endo-1,4-beta-mannanase (EC 3.2.1.78) from Aspergillus niger gave a mixture of oligosaccharides, three of which (II, III, IV) accounted for more than 80% of the GGM. Structural characterisation of these oligosaccharides and the original polysaccharide was achieved by linkage analysis, 1D and 2D NMR spectrometry and enzymatic hydrolysis. Oligosaccharide II beta-D-Glcp-(1-->4)-beta-D-Manp-(1-->, III beta-D-Glcp-(1-->4)-[alpha-D-Galp-(1- >6)]-beta-D-Manp-(1-->, and IV beta-D-Glcp-(1-->4)-[beta-D-Galp-(1-->2)-alpha-D Galp-(1-->6)]-beta-D-Manp-(1-->4)-beta-D-Glcp-(1-->4)-beta-D-Manp-(1-->, appeared in the molar ratio of 2:1:1. A trace amount of mannobiose (I) was detected, indicating that some of the mannosyl residues were contiguous. It is concluded that the predominant structural feature of kiwifruit GGM is a backbone of alternating beta-(1-->4)-linked D-glucopyranosyl and D-mannopyranosyl residues, with approximately one third of the latter carrying side-chains at 0-6 of single alpha-D-Galp-(1--> residues (50% of the branches) or the disaccharide beta-D-Galp (1-->2)-alpha-D-Galp-(1--> (50% of the branches), the substituted residues being separated by three or five unsubstituted monosaccharide units. PMID- 11383900 TI - Further evidence for the gelation ability-structure correlation in sugar-based gelators. AB - Eight methyl glycosides of 4,6-O-benzylidene derivatives of the monosaccharides D glucose, D-mannose, D-allose and D-altrose were synthesized to systematically study the effect of small configurational changes on the ability to gelate organic solvents. Among the beta anomers, only the D-mannose glycoside exhibits a strong gelation ability, whereas in the alpha-series the D-glucose and D-mannose derivatives act as versatile gelators. Also, as a general rule we found that the beta anomers possess a higher ability to gelate solvents than the alpha anomers. The gelation properties are discussed on the basis of SAXS, FTIR, differential scanning calorimetric (DSC) measurements and scanning electron microscopy (SEM) observations. The temperature-dependent SAXS measurements were carried out to elucidate the sol-gel transition temperature. The present study emphasizes that the saccharide family provides, not only valuable information of the structural requirements for the design of new gelators, but also for molecular assembly systems in general. PMID- 11383901 TI - Molecular and crystal structures of N-arylglycopyranosylamines formed by reaction between sulfanilamide and D-ribose, D-arabinose and D-mannose. AB - The X-ray crystal structures of three monosaccharide derivatives prepared by the reaction of sulfanilamide with D-ribose, D-arabinose, and D-mannose have been determined. The derivatives are N-(p-sulfamoylphenyl)-alpha-D-ribopyranosylamine (1), N-(p-sulfamoylphenyl)-alpha-D-arabinopyranosylamine (2), and N-(p sulfamoylphenyl)-beta-D-mannopyranosylamine monohydrate (3). The monosaccharide ring of 1 and 2 has the 1C4 conformation, stabilized in 1 by an intramolecular hydrogen bond from 0-2 to 0-4. Compound 3 has the 4C1 conformation at the monosaccharide ring and the gt conformation at the C-6-O-6 side chain. Occupancy of the water molecule in the crystal of 3 actually examined was 22%. The degree of interaction between sulfamoyl groups and monosaccharide moieties varies from structure to structure. The packing arrangement of 2 involves hydrogen bonding between sulfamoyl groups and monosaccharide hydroxyl groups, but interactions of this type are fewer in 1, and in 3 the hydrogen bonds are either strictly between monosaccharide hydroxyl groups or strictly between sulfamoyl groups. Pairs of hydrogen bonds (two-point contacts) link neighboring molecules in all three structures, between screw-axially related molecules in 1 and 2 and between translationally related molecules in 3. The contact in 3 defined by the O-3-H...O 5 and O-6-H...O-4 hydrogen bonds is found in several other N-aryl-beta-D mannopyranosylamine crystal structures and is apparently an especially favorable mode of intermolecular interaction in these compounds. PMID- 11383902 TI - The anomalous reactivity of the bis(dibutylstannylene) acetal of pentaerythritol: a case of triple activation. AB - The only dibutyltin derivative of pentaerythritol which is observed by refluxing with dibutyltin oxide in methanol is a bis(dibutylstannylene) acetal. This is converted to the expected dibenzyl ether with benzyl bromide, in the presence of tetraethylammonium bromide in boiling toluene, but benzoylation at room temperature gives a tribenzoate. A mechanism is suggested to account for this triple activation. PMID- 11383903 TI - Structure of the O-specific polysaccharide isolated from the lipopolysaccharide of Citrobacter gillenii serotype O12a, 12b strain PCM 1544. AB - A neutral O-specific polysaccharide was isolated from the lipopolysaccharide of Citrobacter gillenii strain PCM 1544, representing serotype O12a,12b. The polysaccharide was studied by sugar and methylation analyses and Smith degradation along with 1H and 13C NMR spectroscopy, including a ROESY experiment. The following structure of the tetrasaccharide repeating unit was established, in which substitution with terminal GlcNAc is approximately 60%. [structure: see text] PMID- 11383904 TI - Investigating the nature of branching in pectin by atomic force microscopy and carbohydrate analysis. AB - Atomic force microscopy (AFM) has been used to investigate the nature of the long branches attached to pectin which were described in a previous report [Round, A. N.; MacDougall, A. J.; Ring, S. G.; Morris, V. J. Carbohydr. Res. 1997, 303, 251 253]. Analysis of the AFM images and comparison with neutral sugar and linkage analyses of the two pectin fractions suggest that the distribution and total amount of branches observed do not correspond with the pattern of neutral sugar distribution. It is thus postulated that the long chains consist of polygalacturonic acid, attached via an as yet undetermined linkage to the pectin backbone, with the neutral sugars present as short, undetected branches. This explanation would have important implications for the nature of 'in situ' pectin networks within plant cell walls and models of gelation in commercial extracted pectin, and the existence of significant branching will markedly influence the viscosity of extracted pectins. PMID- 11383905 TI - Preparation, single-crystal X-ray diffraction and high-resolution NMR spectroscopic analyses of N-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl) trimethylammonium bromide. AB - Preparation of N-(2,3,4,6-tetra-O-acetyl-beta-D-glucopyranosyl)trimethylammonium bromide as the unique N-glycosylated quaternary salt derived from trialkylamine is described. The structure of the compound was elucidated by 1H and 13C NMR spectroscopy and by single-crystal X-ray analysis as well. PMID- 11383906 TI - GH/IGF-I normalization and tumor shrinkage during long-term treatment of acromegaly by lanreotide. AB - New depot somatostatin analogs such as lanreotide-slow release (LAN) represent a significant improvement in the medical treatment of acromegaly. Seventy-three consecutive acromegalic patients, treated by LAN, were evaluated in a retrospective monocentric study. Sixteen were excluded from further evaluation due to combined treatment with dopamine agonist drugs, early LAN withdrawal for persistence of headache, or gastrointestinal side-effects. Fifty-seven patients (aged 20-82 years, 16 males) were thus evaluated. Thirty-two patients had been previously treated by neurosurgery (Tx) and/or radiotherapy (Rx). After washout, LAN (30 mg) was administered im at 10-14-day intervals. Time intervals between injections were then individually tailored to normalize IGF-I levels. LAN was administered for 12 (6-36) [median (range)] months. GH and IGF-I levels decreased from 13 (7-20) [median (interquartile)] microg/l to 3.2 (1.7-6.2) microg/l (p<0.0001) and from 780 (596-1000) microg/l to 264 (180-530) microg/l (p<0.000001), respectively. Seven patients were resistant to treatment. Among the 50 sensitive patients, GH levels fell below 2.5 microg/l in 52% (and below 1 microg/l in 18%), IGF-I levels normalized in 72% and both results were obtained in 46%. IGF-I values normalized in 87% of patients treated every 14 days, in 100% every 21-28 days, in 69% every 10 days and in 22% every 7 days. No different control of GH/IGF-I hypersecretion was evidenced between patients previously treated or not by Tx and/or Rx. Patients with the lowest basal hormonal levels and those over 55 years showed greater responsiveness (both p<0.05). The maintenance of LAN schedule up to 18 months determined a further suppression (p=0.04 for IGF-I). A reduction of tumor size was shown in 60% of evaluated patients (6/10). HbA1c slightly increased in 42% of patients and gallstones were observed in 16%. LAN is a very effective tool in the treatment of acromegaly: its chronic administration normalizes GH/IGF-I levels in most patients, shrinks the tumor in a high percentage of patients and seems to control hormonal hypersecretion as primary treatment as well as neurosurgery. PMID- 11383907 TI - Lack of evidence of a genetic origin in the impaired spermatogenesis of a patient cohort with low-grade varicocele. AB - Varicocele is the most common clinical finding in infertile men but controversy continues to surround the utility of its treatment. An increased response of FSH to gonadotrophin-releasing hormone testing has been described in patients with varicocele, while the co-influence of Yq chromosome microdeletions in the infertility associated to this pathology is still under investigation. We studied 30 patients with first- and second-grade varicocele, 15 idiopathic oligozoospermic men and 21 age-matched healthy controls. All subjects underwent testicular Doppler ultrasonography, semen analysis, gonadotrophin-releasing hormone testing and baseline blood sampling for total and free testosterone, PRL, 17beta-estradiol, SHBG evaluation and Yq chromosome analysis. Apart from FSH, no difference in baseline hormonal levels was found between the groups. The patients with varicocele showed both an increased basal (p=0.007) and GnRH-induced FSH response (peak and AUC) (p=0.004) in comparison with the controls, while the idiopathic oligozoospermic men had only higher GnRH-induced FSH AUC (p=0.04). In the varicocele group, FSH peaks after GnRH testing correlated positively with the grade of disease (r=0.42, p=0.02) and negatively with sperm count (r=-0.50, p=0.005) and bilateral testis volume (r=-0.52, p=0.005). Sperm count and sperm motility were similarly significantly reduced both in patients with varicocele and in patients with idiopathic oligozoospermia in comparison with healthy controls. Yq chromosome analysis by sequence-tagged site PCR revealed no microdeletion in the AZF regions in any subject studied. Given the quite small number of subjects studied, our overall findings can only prompt us to suggest a possible causal role of varicocele in the impairment of spermatogenesis in our patients. Furthermore, although a genetic co-influence (i.e. Yq microdeletions) does not seem to be involved in the pathogenesis of infertility in men with varicocele and mild to moderate oligozoospermia, genetic screening seems to be advisable, especially in those patients who present a severe impairment of sperm count, as has been suggested by recent literature data. PMID- 11383908 TI - Effects of 12-month GH treatment on bone metabolism and bone mineral density in adults with adult-onset GH deficiency. AB - Serum bone-Gla protein (BGP), bone alkaline phosphatase (B-AP), and C-terminal cross-linked telopeptide of type I collagen (ICTP) levels were evaluated in 18 adults with acquired GH deficiency (GHD, 14 males and 4 females, age range: 25-59 yr) before, at 3, 6, 9 and 12 months of rec-GH treatment (0.125 IU/kg/week for the first month, followed by 0.25 IU/kg/week for 11 months) and 6 months after the withdrawal of therapy. Total body bone mineral density (BMD, g/cm2) was measured with dual energy X-ray absorptiometry (Hologic QDR 1000/W) before, at 12 months of GH treatment and 6 months after its withdrawal. Before treatment, BGP (mean+/-SE: 5.1+/-0.4 ng/ml), B-AP (59.4+/-6.5 IU/l), ICTP (3.1+/-0.3 ng/ml) levels of patients were similar to in healthy controls (BGP: 5.4+/-0.1 ng/ml; B AP: 58.2+/-2.0 IU/l; ICTP: 4.1+/-0.3 ng/ml). GH treatment caused a significant increase of BGP, B-AP, ICTP levels, the maximal stimulation of bone resorption, occurring after 3 months of GH treatment, while the maximal effect on bone formation being evident later (at 6th month). A slight decline in BGP, B-AP, T-AP and ICTP levels occurred at 9-12 months of therapy, although the values remained significantly higher than in basal conditions and with respect to healthy controls. Before treatment, mean total body BMD of patients (1.110+/-0.027 g/cm2, range: 0.944-1.350 g/cm2) was not significantly different (z-score: +0.47+/-0.31, NS) from that observed in healthy controls (1.065+/-0.008 g/cm2, range: 1.008 1.121 g/cm2). GH therapy was associated with a significant reduction of mean total body BMD values (6th month: -1.8+/-0.5%, p<0.01; 12th month: -2.1+/-1.0%, p<0.05 vs baseline), particularly evident in the first six months of treatment. Six months after the withdrawal of GH therapy, BGP (5.9+/-0.5 ng/ml), B-AP (57.3+/-7.0 IU/l) and ICTP (3.2+/-0.1 ng/ml) levels returned similar to those recorded before treatment, while total BMD increased (+1.5+/-0.7, p<0.05), remaining however slightly lower than in basal conditions (-0.6+/-1.2, NS). In conclusion, our study shows that: a) acquired GHD in adulthood is associated with both normal bone formation/resorption indexes and normal total body BMD; b) GH therapy causes a significant rise of bone formation/resorption markers (earlier and greater for bone resorption); c) one-year GH therapy is associated with a reduction of total body BMD values, particularly evident in the first 6 months of treatment; d) the effects of GH therapy on bone turnover are transient, being completely reverted six months after the withdrawal of GH therapy; e) the increase of total body BMD (up to baseline values) after GH withdrawal might be explained as consequence of persisting effects of previous GH stimulation on bone remodeling. PMID- 11383909 TI - Effect of acute hyperglycemia on potassium (86Rb+) permeability and plasma lipid peroxidation in subjects with normal glucose tolerance. AB - Hyperglycemia is likely to be one of the important determinants of ion transport as it is known to induce oxidative stress and may thus enhance non-specific permeability of membranes. The aim of the present study was to evaluate the effects of an acute increase in glycemia on 86Rb+ (a marker for K+) influx and lipid peroxidation. We evaluated the 75-g oral glucose tolerance test (OGTT) induced modification on 86Rb+ influx and plasma lipid peroxidation in 20 subjects with normal glucose tolerance (NGT). After 2-hour glucose loading, the levels of passive 86Rb+ influx and plasma lipid peroxidation were significantly increased, whereas the active influx of 86Rb+ was unchanged. The total and passive influx of 86Rb+ into erythrocytes was significantly correlated with the level of plasma lipid peroxidation. This study demonstrates that acute hyperglycemia induces an increase in the passive influx of 86Rb+ in subjects with NGT, suggesting that acute hyperglycemia may produce an oxidative stress in plasma. These changes may be among the earliest changes occurring in response to hyperglycemia. PMID- 11383910 TI - An assessment of genetic markers as predictors of bone turnover in healthy adults. AB - In 1992 a significant relationship between bone turnover and the vitamin D receptor (VDR) genotype was reported in Australian subjects of UK-Irish decent. Since then, several groups have investigated the relationship between VDR and other bone-related genotypes, bone mass and bone turnover in several populations. However, the results of these studies are conflicting. Therefore, our aim was to determine bone-related genotypes in a population of healthy Irish adults and relate these genotypes to the rate of bone turnover. One hundred and eighteen healthy Irish adults (aged 19-67 yr) were recruited and fasting blood and first void urines were collected from each subject. Bone-related genotype frequencies in healthy Irish adults were similar to those reported in other Caucasian populations and were in Hardy-Weinberg equilibrium. Estrogen receptor (Pvu II or Xba I), apolipoprotein E and collagen IA1 genotypes were not related to bone turnover. The tt VDR genotype was associated with significantly higher serum osteocalcin (29% and 40%) compared with the Tt and TT genotypes, respectively. The ff VDR genotype was associated with significantly higher urinary pyridinoline (by approximately 44% and approximately 29%) and deoxypyridinoline (by approximately 76% and approximately 58%) levels and higher serum osteocalcin (by approximately 25% and approximately 53%) compared with the Ff or FF genotypes, respectively. These findings suggest that healthy Irish adults with either the tt or ff VDR genotype have higher rates of bone turnover than those with Tt or TT, or Ff or FF genotypes, respectively, and therefore may have a higher risk of low bone mineral density and osteoporosis in later life. PMID- 11383911 TI - Bone mineral density and bone markers in hypogonadotropic and hypergonadotropic hypogonadal men after prolonged testosterone treatment. AB - After prolonged treatment (76.4+/-10 and 70.1+/-12.3 months, respectively) (mean+/-SE) with testosterone enanthate (250 mg i.m. every 3 weeks), bone mineral density (BMD) and bone metabolism were evaluated in 12 patients (aged 29.3+/-1.4 yr) affected by idiopathic hypogonadotropic hypogonadism (IHH), in 8 patients (29.6+/-2.6 yr) affected by Klinefelter's syndrome (KS), and in 10 healthy men (30.6+/-1.7 yr) matched according to age and BMI. Spinal BMD in IHH was significantly lower than in controls (0.804+/-0.04 vs 1.080+/-0.01 g/cm2; p<0.001), while there was no difference in neck BMD (0.850+/-0.01 vs 0.948+/-0.02 g/cm2). Neither spinal (0.978+/-0.05 g/cm2) nor neck (0.892+/-0.03 g/cm2) BMD in KS were significantly different from controls. Six IHH and one KS subjects were osteoporotic, while 6 IHH and 2 KS subjects were osteopenic. A significant inverse correlation was found between spinal BMD and age at the treatment onset in IHH (r=-0.726, p=0.007). In IHH there were significant increases in bone formation (alkaline phosphatase=318.3+/-33.9 vs 205.4+/-20.0 IU/l; osteocalcin=13.44+/-1.44 vs 8.57+/-0.94 ng/ml; p<0.05) and in bone resorption (urinary cross-linked N-telopeptides of type I collagen=149.1+/-32.3 vs 47.07+/ 8.4 nmol bone collagen equivalents/mmol creatinine; p<0.05) compared to controls, while such differences were not present in KS. Our results outline the importance of BMD evaluation in all hypogonadal males. Nevertheless, bone loss is a minor characteristic of KS, while it is a distinctive feature of IHH. Therefore, early diagnosis and age-related replacement therapy coupled with a specific treatment for osteoporosis could be useful in preventing future severe bone loss and associated skeletal morbidity. PMID- 11383912 TI - Type I 5'-iodothyronine deiodinase activity and mRNA are remarkably reduced in renal clear cell carcinoma. AB - The purpose of this study was to compare thyroid hormone metabolism between non cancerous tumor-surrounding human kidney tissues and renal clear cell carcinomas (RCCC). The material consisted of samples taken from 10 RCCC patients of both sexes and three grades of differentiation, G1 to G3. We showed that, similar to rat tissue, type I 5' monodeiodinase (5'DI) expression is heterogeneous within the human kidney. We also found a poor correlation between 5'DI activity and mRNA level in non-cancerous tumor-surrounding tissue suggesting significant post transcriptional regulation of 5'DI expression by an unidentified process in the human kidney. In all RCCC tissues both 5'DI activity and mRNA levels were undetectable. This suggests either loss of human 5'DI gene expression during neoplastic transformation or the origination of RCCC from a tubular cell type that does not express 5'DI. PMID- 11383913 TI - Lymphocytic hypophysitis: its expanding features. AB - Lymphocytic hypophysitis is classically defined as an inflammatory disorder confined to adenohypophysis. However, it has recently been indicated that infundibuloneurohypophysitis underlies some subsets of central diabetes insipidus (DI). Therefore, lymphocytic hypophysitis can be considered a syndrome including disorders of both the anterior pituitary (lymphocytic adenohypophysitis) and the posterior pituitary (lymphocytic infundibuloneurohypophysitis). We describe a 77 yr-old woman with lymphocytic hypophysitis presenting with headache, diplopia, general malaise and appetite loss. Head magnetic resonance imaging (MRI) demonstrated pituitary swelling and dura mater thickening on the dorsum sella. Endocrinological investigations revealed both anterior and posterior pituitary dysfunction associated with primary hypothyroidism due to Hashimoto's thyroiditis. Headache and diplopia spontaneously disappeared, and anterior pituitary dysfunction, general malaise and appetite loss improved after taking 10 mg hydrocortisone daily, although ACTH hyposecretion persisted. Pituitary swelling was thereafter reduced but the dura mater thickening persisted. We suggest that this case may represent a variant of lymphocytic hypophysitis in which chronic inflammatory process involves both the anterior and the posterior pituitary gland, infundibulum, dura mater on the dorsum sella and cavernous sinus. Regarding expanding features of lymphocytic hypophysitis, it may be considered a syndrome including heterogeneous disorders, of which the pathogenesis remains to be elucidated. PMID- 11383914 TI - Conn's syndrome and bilateral renal artery stenosis in the presence of multiple renal arteries. AB - We report the case of a 42-year-old male who was admitted to our hospital after an acute hypertensive crisis despite four-way anti-hypertensive therapy. The renal scintigraphy, the excretory urogram and the biochemical profile performed two years before were unremarkable, except for slightly elevated serum creatinine and plasma aldosterone, in presence of normal aldosterone/renin ratio. The renal arterial angiography that was performed despite a second unremarkable scintigraphy revealed high-grade bilateral arterial stenosis in the presence of multiple renal arteries. Following dilatation of the left stenosis, the aldosterone/renin ratio was pathologic. Recumbent and orthostatic aldosterone values were 830 pg/ml and 1824 pg/ml, respectively, and recumbent and orthostatic renin values were 0.82 and 1.21 ng angiotensin I/ml/h, respectively. The abdominal computed tomography performed to investigate a possible concomitant Conn's syndrome resulted in the detection of a left adrenal tumor. After resection of the lesion, plasma-aldosterone levels normalized and a pronounced rise in serum potassium levels was observed. Following angioplasty of the right renal artery stenosis, blood pressure could easily be managed with combined beta and calcium channel blocker therapy. Particularly in cases of bilateral (but also in the presence of unilateral) renal artery stenosis in association with Conn's syndrome, all the available screening methods for these disorders can fail. In cases of poor response to combination hypertensive therapies, renal arteriography and a fludrocortisone-suppression test should be performed in order to rule out both renal arterial stenosis and Conn's syndrome, even in the absence of clinical and biochemical findings suspicious for either disorder. PMID- 11383915 TI - Enhancing insulin action: from chemical elements to thiazolidinediones. AB - Type 2 diabetes is characterized by two fundamental biological defects: a reduced glucose-dependent insulin secretion and an increased resistance to the action of insulin at the level of various target tissues. While the use of agents to improve the insulin secretory activity of the islets of Langerhans has witnessed the flourishing of several new drugs over the years, a much greater difficulty has been experienced in the search for insulin-sensitizing drugs. The aim of this article is to critically review this topic, and to emphasize the importance of providing alternative strategies for the management of Type 2 diabetes. PMID- 11383916 TI - Bone mineral density in acromegaly: does growth hormone excess protect against osteoporosis? PMID- 11383917 TI - White addison's disease: an intrinsic cellular defect. PMID- 11383918 TI - "Madonna with the infant". Unknown painter, XII-XIV century. PMID- 11383919 TI - Different insulin-like 3 (INSL3) gene mutations not associated with human cryptorchidism. AB - Cryptorchidism is the most frequent congenital anomaly of the urogenital tract in the male, but its etiology is for the most part unknown. Evidence suggests that a possible genetic cause may be involved. Animal models support this hypothesis, and in particular INSL3 (Leydig insulin-like 3 hormone) has been proposed as putative gene for cryptorchidism, since male mice mutant for Insl3 exhibit bilateral abdominal cryptorchidism due to alteration of gubernaculum development. In this study, we analyzed whether mutations in INSL3 could be associated with human cryptorchidism. Heteroduplex analysis and sequencing of both exons of INSL3 in 65 ex-cryptorchid patients and a group of control subjects allowed us to find four nucleotide changes in the sequence of exon I. These mutations are all single base substitutions from G to A at position 27, 96, 126 and 178. Only the 178G-->A substitution changes codon 60 from alanine to threonine (A60T). All mutations were found in comparable distribution in ex-cryptorchid patients and non cryptorchid men. Therefore, all mutations represent neutral polymorphisms not associated with phenotype. This study confirms previous observations and demonstrates a novel polymorphism in the INSL3 gene. In contrast to that described for the mutant mouse, these data indicate that mutations of INSL3 do not seem to represent a frequent cause of cryptorchidism. PMID- 11383920 TI - Effects of angiotensin II on adipose conversion and expression of genes of the renin-angiotensin system in human preadipocytes. AB - A complete functional renin-angiotensin system exists in human adipose tissue, but its regulation and the effects of angiotensin II on cells from this tissue are only beginning to be understood. In this study, we examined the effects of angiotensin II on changes in lipid accumulation, specific glycerol-3-phosphate dehydrogenase activity, and the expression of five genes of the renin-angiotensin system during the adipose conversion of human primary cultured preadipocytes. Angiotensin II leads to a distinct reduction in insulin-induced differentiation, but only has a marginal effect on the adipose conversion of cells stimulated with insulin, cortisol, and isobutyl methyl xanthine. During differentiation, angiotensinogen mRNA levels rise, renin mRNA levels decline, whereas renin binding protein and angiotensin-converting enzyme levels are unaffected. Angiotensin II downregulates angiotensinogen and renin gene expression, but it does not affect renin-binding protein and angiotensin-converting enzyme levels. Angiotensin II thus prevents the development of adipocytes in contact with high insulin levels, while not inhibiting differentiation, which is further stimulated. Therefore, angiotensin II could be a protective factor against uncontrolled expansion of adipose tissue. Further studies are needed to find out whether the effects of angiotensin II on the renin-angiotensin system are direct feedback loops or secondary to changes in the differentiation program. PMID- 11383921 TI - Effect of angiotensin peptides on PAI-1 expression and production in human adipocytes. AB - Angiotensin (Ang) II is the active component of the renin-angiotensin-system (RAS), but its degradation products have also been shown to exhibit biological activity. This system, which mainly controls blood pressure and electrolyte homeostasis, was recently found to be completely expressed in human adipose tissue. The major determinant in the fibrinolytic system is the plasminogen activator inhibitor-1 (PAI-1). Both PAI-1 and components of the RAS are over expressed in the obese state. We have recently shown that Ang II is able to induce PAI-1 expression and release via the AT1-receptor in human fat cells in primary culture, and have provided the first evidence that two metabolites, Ang III and Ang IV, may have a similar stimulatory effect on PAI-1 release. We have now performed additional experiments to further characterize the role of the angiotensin peptides in the production of PAI-1. Ang III and Ang IV showed a time and dose-dependent stimulation of PAI-1 protein release. Concomitantly, mRNA levels were markedly elevated. Using specific receptor blockers, all angiotensin peptides seem to induce PAI-1 expression via the angiotensin receptor subtype 1. However, components of the renin-angiotensin-system seem to play an important role in the control of fibrinolysis in adipose tissue. We conclude that PAI-1 production by adipose tissue may contribute to the elevated thromboembolic risk in obesity. PMID- 11383922 TI - Protein kinase C alpha modulates the Ca2+ influx phase of the Ca2+ response to 1alpha,25-dihydroxy-vitamin-D3 in skeletal muscle cells. AB - Treatment of chick skeletal muscle cells with 1alpha,25-dihydroxy-vitamin D3 [1alpha,25(OH)2D3] triggers a rapid and sustained increase in cytosolic Ca2+ ([Ca2+]i), which depends on Ca2+ mobilization from inner stores and extracellular Ca2+ entry. Fluorimetric analysis of changes in [Ca2+]i in Fura-2-loaded cells revealed that the hormone significantly stimulates the Ca2+ influx phase within the concentration range of 10(-12)-10(-6) M, with maximal effects (3.5-fold increase) at 10(-9) M 1alpha,25(OH)2D3. The effects of the sterol on the Ca2+ entry pathway were abolished by the PKC inhibitors bisindolylmaleimide and calphostin. We have recently shown that, in these cells, 1alpha,25(OH)2D3 activates and translocates PKC alpha to the membrane, suggesting that this isozyme accounts for PKC-dependent 1alpha,25(OH)2D3 modulation of Ca2+ entry. The role of PKC alpha was specifically addressed here using antisense technology. When the expression of PKC alpha was selectively knocked out by intranuclear microinjection of an antisense oligonucleotide against PKC alpha mRNA, the Ca2+ influx component of the response to 1alpha,25(OH)2D3 was markedly reduced (-60%). These results demonstrate that 1alpha,25(OH)2D3-induced activation of PKC alpha enhances extracellular Ca2+ entry partially contributing to maintainance of the sustained phase of the Ca2+ response to the sterol. PMID- 11383923 TI - Effect of leptin on insulin, glugacon and somatostatin secretion in the perfused rat pancreas. AB - We have investigated the effect of rat leptin as well as the 22-56 fragment of this molecule on pancreatic hormone secretion in the perfused rat pancreas. In pancreases from fed rats, leptin failed to alter the insulin secretion elicited by glucose, arginine or tolbutamide, but inhibited the insulin response to both CCK-8 and carbachol, secretagogues known to act on the B-cell by increasing phospholipid turnover. This insulinostatic effect was also observed with the 22 56 leptin fragment. In pancreases obtained from 24-hour fasted rats, no effect of leptin on carbachol-induced insulin output was found, perhaps as a consequence of depressed B-cell phospholipid metabolism. Leptin did not influence glucagon or somatostatin release. Our results do not support the concept of leptin as a major regulator of B-cell function. Leptin inhibition of carbachol-induced insulin output might reflect a restraining effect of this peptide on the cholinergic stimulation of insulin release. PMID- 11383924 TI - Extraction of total RNA from adipocytes. AB - RNA isolation from adipocytes presents with several technical problems and yields unacceptable results when following standard protocols. Here, we will describe additional steps and modifications necessary for the use of different RNA isolation protocols in terms of RNA yield, RNA quality and preparation time. Using five times the recommended quantity of lysis buffer, incubating the lysate at 37 degrees C, repeatedly passing the lysate through a cannula, and centrifugation to remove the lipid layer are essential additional steps when working with adipocytes. With these modifications, isolation of total RNA resulted in an average yield of 12-30 microg total RNA from 2 x 10(6) cells. Preparation times were similar for all but the CsCl gradient method. The purest RNA was obtained by spin-column purification, whereas acid phenol-chloroform methods yielded the highest amounts of total RNA. CsCl gradient ultracentrifugation is suggested for situations where DNase I digestion is impractical. PMID- 11383925 TI - Hyperprolactinemia and luteal insufficiency in infertile patients with mild and minimal endometriosis. AB - The objective of the present paper was to assess the presence of hormonal alterations in infertile women with stage I or II endometriosis (Group III, n = 20) compared to fertile women without endometriosis (Group I, n = 14) and to fertile women with endometriosis (Group II, n = 7). Serum levels of FSH, LH, estradiol, TSH, and PRL were measured between days 1 and 5 of the early follicular phase; in the luteal phase, three serum samples were collected for progesterone measurement, and endometrial biopsies were performed. Serum estradiol levels were lower (p = 0.035) in infertile patients with endometriosis than in fertile patients without endometriosis. Six infertile patients with endometriosis presented prolactin levels above 20 ng/ml. This was not observed in the other groups. Luteal insufficiency was more frequent in infertile patients with endometriosis (78.9%) than in fertile patients with (42.9%) or without endometriosis (0%). In a multiple logistic regression analysis, only the presence of endometriosis and infertility was significantly associated with luteal insufficiency. The serum levels of LH, FSH, and TSH were not significantly different among the groups. Luteal insufficiency and altered prolactin secretion were associated with endometriosis, and could be important mechanisms causing infertility in this group of patients. PMID- 11383927 TI - Lipid peroxidation, antioxidant defence and acid-base status in cord blood at birth: the influence of diabetes. AB - Pregnancy complicated by poor control of diabetes is associated with a higher risk of embryopathies, spontaneous abortions and perinatal mortality. A number of authors suggest an involvement of reactive oxygen species (ROS) in diabetic pregnancy. Determining lipid peroxidation products (LP), scavenging enzyme activities and the umbilical cord blood's acid-base balance may contribute to an adequate diagnosis of the neonate at birth. Nevertheless, such measurements seem to have limited value in practical clinical routine. The present study evaluates LP, antioxidant defence and acid-base status related to diabetic pregnancy. Twenty-eight women with type 1 diabetes (PGDM), 19 with gestational diabetes (GDM) and 13 control cases were investigated. An additional control group consisted of 15 healthy patients with negative diabetic history; all women underwent vaginal delivery. Immediately after delivery cord blood samples and placental tissue were collected for malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione (GSH) determination. Additionally, pH, pCO2, pO2 and base excess were measured in both vessels and compared to identify and exclude double venous samples. MDA levels in both cord blood and placental homogenates were significantly higher in both pregestational and gestational diabetic groups, but SOD activity was significantly diminished. Cord blood GSH was markedly elevated in PGDM and GDM. We have also shown significant differences in acid-base parameters in infants of PGDM group. Statistical analysis was performed using the Mann-Whitney U-test. These findings indicate an excessive oxidative stress in pregnancy complicated by diabetes mellitus. Evaluating LP products and scavenging enzyme activities may be valuable, sensitive indexes of fetal/neonatal threat in diabetic pregnancy in humans. Since oxidative stress is an important pathway for fetal injury, we believe that obtaining adequate measurements at the time of birth would contribute to clarifying the fetal/neonatal status in a medical and legal context and might be of value in altering therapy in newborn infants. PMID- 11383926 TI - Absence of mutations in the coding regions of follicle-stimulating hormone receptor gene in Singapore Chinese women with premature ovarian failure and polycystic ovary syndrome. AB - Normal gonadal function is critically dependent on the integrity of pituitary gonadal axis, where follicle-stimulating hormone (FSH) plays a key role. In the female, FSH is required for follicular growth, estrogen production and oocyte maturation. Its function is mediated by its specific receptor (FSHR), and defective FSHR has been shown to affect folliculogenesis and ovarian function. In this study, we screened the entire coding region of FSHR gene for pathogenic mutations in women with premature ovarian failure (POF) (n = 16) and polycystic ovary syndrome (PCOS) (n = 124) and found no mutations in these patients. Two known polymorphisms, Thr307Ala and Ser680Asn showed similar distributions of the allelic variations and protein isoforms in PCOS and normal control subjects (n = 236). It appears from this study that mutations in the coding regions of FSHR gene are not a causative factor of the above clinical manifestations in Chinese Singapore women. PMID- 11383928 TI - TSBAb (TSH-stimulation blocking antibody) and TSAb (thyroid stimulating antibody) in TSBAb-positive patients with hypothyroidism and Graves' patients with hyperthyroidism. AB - There are two types of TSH receptor antibodies (TRAb); thyroid stimulating antibody (TSAb) and TSH-stimulation blocking antibody (TSBAb). TSAb causes Graves' hyperthyroidism. TSBAb causes hypothyroidism. Both TSAb and TSBAb block TSH-binding to thyroid cells as TSH receptor antibodies (TRAb). TSBAb-positive patients with hypothyroidism and Graves' patients with hyperthyroidism may have both TSBAb and TSAb. We studied TSBAb and TSAb in 43 TSBAb-positive patients with hypothyroidism and in 55 untreated Graves' patients with hyperthyroidism. TSBAb activities were expressed as percentage inhibition of bovine (b) TSH-stimulated cAMP production by test IgG. Two formulas were used to calculate TSBAb activities; TSBAb-A (%) = [1 - (c - b)/(a - b)] x 100 and TSBAb-B (%) = [1 - (c - d)/(a - b)] x 100, where a: cAMP generated in the presence of normal IgG and bTSH, b: cAMP generated in the presence of normal IgG, c: cAMP generated in the presence of test IgG and bTSH, and d: cAMP generated in the presence of test IgG. TSAb (%) = [d/b] x 100. All of the 43 TSBAb-positive patients with hypothyroidism had strongly positive TSBAb-A and -B. Some of them had weakly positive TSAb (<240%). All 55 untreated Graves' patients had positive TSAb (205-2509%). Some of them had both TSAb and TSBAb. TSBAb-positive patients with hypothyroidism had a limited distribution of TSBAb- and TSAb-activities (TSBAb-A + 75 - + 103%, TSBAb B + 87 - + 106%, TSAb 92-240%), but Graves' patients with hyperthyroidsim had a wide distribution of TSAb- and TSBAb-activities (TSAb 205-2509%, TSBAb-A - 158 - + 43%, TSBAb-B - 14 - + 164%). TSBAb-A ignores TSAb activity in serum, and might give low TSBAb activity. However, TSBAb-A clearly differentiates TSBAb-positive patients with hypothyroidism from Graves' patients with hyperthyroidism; thus, we favor TSBAb-A over TSBAb-B. Some of TSBAb-positive patients with hypothyroidism and Graves' patients with hyperthyroidism have both TSBAb and TSAb. PMID- 11383929 TI - Acute plasma volume expansion in the untrained alters the hormonal response to prolonged moderate-intensity exercise. AB - To investigate the role of an increase in plasma volume (PV), characteristically observed with short-term endurance training, on the endocrine response to prolonged moderate intensity exercise, eight untrained males (VO2 peak = 3.52 +/- 0.12 l x min(-1)) performed 90 min of cycle ergometry at approximately 60% VO2peak both before (CON) and following (PVX) PV expansion. Acute PV expansion, which was accomplished using a solution of Dextran (6%) or Pentispan (10%) (6.7 ml kg(-1)), resulted in a calculated 15.8+/-2.2% increase (p<0.05) in PV. The prolonged exercise resulted in increases (p<0.05) in plasma vasopressin (AVP), plasma rennin activity (PRA), aldosterone (ALD), atrial naturetic peptide (alpha ANP), and the catecholamines norepinephrine (NE) and epinephrine (EPI). PVX blunted the increases (p<0.05) in AVP, PRA, ALD, NE and EPI, during the exercise itself. The concentration of alpha-ANP was also lower (p<0.05) during exercise following PVX, an effect that could be attributed to the lower resting levels. No differences in osmolality was observed between conditions. These results demonstrate that PVX alters the fluid regulatory hormonal response in untrained subjects to moderate intensity dynamic exercise in a manner similar to that observed following short-term training induced alterations in PV. The specific mechanisms responsible for these alterations remain unclear, but appear to be related directly to the increase in PV. PMID- 11383930 TI - Leptin in type 2 diabetic or myotonic dystrophic women. AB - BACKGROUND: Insulin resistance is an important determinant of circulating leptin concentrations in humans, but its independent contribution on plasma leptin levels are controversial. In the present study, we characterized plasma leptin levels and their regulation in women with 2 different insulin resistance states: type 2 diabetes and myotonic dystrophy disease, and in controls. MATERIAL AND METHODS: We studied 3 groups of women: 21 type 2 diabetic patients, 20 myotonic dystrophic patients and a control group of 20 normoglycemic subjects, matched in age and body mass index. Body composition, fasting glucose and insulin, IGF-I, IGF-binding protein-3 and leptin were studied. Body composition was measured using a bioelectrical impedance analyser. Insulin sensitivity (in percentage) was modeled according to a computer-based homeostasis model assessment model. Data are expressed in mean +/- SEM. RESULTS: In both groups of patients, glucose concentrations were higher in type 2 diabetic patients than in myotonic dystrophic patients, and insulin concentrations and insulin sensitivity were similar in the 2 groups of patients (82.4 +/- 18.6% in type 2 diabetic patients vs. 69.7 +/- 9.7% in myotonic dystrophic patients, p = 0.2) and lower than in controls. Serum leptin and leptin/fat mass ratio were higher in myotonic dystrophic patients than in type 2 diabetic patients (30 +/- 4.9 ng/ml vs. 17.7 +/- 2.6 ng/ml, p = 0.03 and 2.32 +/- 0.69 ng/ml/kg vs. 1.07 +/- 0.2 ng/ml/kg, p = 0.02, respectively) or those found in controls. In type 2 diabetic patients, leptin concentrations were correlated with body mass index and body fat, and in myotonic dystrophic patients leptin concentrations were correlated with age, body mass index, fasting insulin and lower insulin sensitivity, whereas leptin concentrations were not correlated with body fat. CONCLUSIONS: These findings suggest that leptin concentrations and regulation in myotonic dystrophic patients are different from type 2 diabetes. PMID- 11383931 TI - Inhibition of cyclooxygenase-1 or -2 on insulin sensitivity in healthy subjects. AB - The aim of this study was to identify the effects of cyclooxygenase (COX)-1 and 2 inhibition each on insulin sensitivity in healthy subjects. A randomized, double-blind, controlled clinical trial was carried out in 21 young, healthy, non obese male volunteers. Pharmacological COX-1 inhibition was performed with the prescription of acetylsalicylic acid (ASA) at a low dose, and COX-2 selective inhibition was performed with celecoxib. After randomization, all subjects received an oral morning dose of ASA 100 mg (n = 7), celecoxib 200 mg (n = 7), or placebo for the control group (n = 7) during a period of 15 days. Before and after of the study period, a metabolic profile was measured in all participants. An insulin tolerance test (ITT) was performed to assess insulin sensitivity, and the constant for the serum glucose disappearance rate (K ITT) was calculated. Clinical and metabolic characteristics were similar between groups. The K ITT calculated with the ITT was higher after celecoxib than at baseline (4.8 +/- 0.9 vs. 4.3 +/- 0.6%/min, p = 0.04), indicating improvement in insulin sensitivity. Neither ASA nor placebo administrations modified insulin sensitivity. In conclusion, COX-2-selective inhibitor at a celecoxib dose of 200 mg daily increased insulin sensitivity in healthy subjects. PMID- 11383932 TI - Tube feeding in stroke patients: a medical and ethical perspective. AB - Dysphagia, or difficulty swallowing, is a relatively common consequence following stroke, with most sources quoting rates of around 40%. The percentage of stroke patients who require tube feeding for nutritional support varies quite widely, with studies quoting rates in the acute phase from 8.5% to 29%. Since stroke patients are likely to constitute a high percentage of patients on a neurology or rehabilitation ward, neurologists and physiatrists are likely to be confronted with the sometimes challenging decision of whether and when to commence tube feeding and whether and when to discontinue it after it has begun. This decision making process is likely to involve medical, ethical and legal considerations and the main purpose of this paper is to review these considerations and provide some practical recommendations. PMID- 11383933 TI - The pediatric neurologist as expert witness with particular reference to perinatal asphyxia. AB - The frequency of litigation related to alleged medical malpractice is increasing in Canada. For the neurologist, involvement in such litigation most often takes place in the context of acting as an expert witness and, for the pediatric neurologist, the most common clinical situation for which expertise is requested is that of possible perinatal asphyxia. The medical expert's primary role is to provide necessary guidance and assistance to the court, which may permit the rendering of decisions that are scientifically valid. This article will review the attributes of the medical expert witness. Aspects of perinatal asphyxia cases under litigation that commonly require the assistance of pediatric neurology expertise such as etiology, timing, extent of disability and life expectancy will also be reviewed in detail. The aim is to provide for the neurologist a clearer understanding of the responsibilities inherent in this increasing professional role. PMID- 11383935 TI - Outpatient craniotomy for brain tumor: a pilot feasibility study in 46 patients. AB - BACKGROUND: Since 1991 the author has routinely performed awake craniotomy for intra-axial brain tumors with low complication rate and low resource utilization. In late 1996 a pilot study was initiated to assess the feasibility of performing craniotomy for tumor resection as an outpatient procedure. METHODS: A rigorous protocol was developed and adhered to, based around the patient's arrival at hospital at 6:00 a.m, undergoing image-guided awake craniotomy with cortical mapping, and being discharged by 6:00 p.m. RESULTS: During the 48 month period from December 1996 to December 2000, 245 awake craniotomies were performed and of those, 46 patients were entered into the outpatient craniotomy protocol. Pathology in the 46 intent-to-treat group was: 21 metastasis, 19 glioma, and six miscellaneous. Four patients required conversion to inpatients and one patient was readmitted later the same evening due to headache. Thus 41/46 patients successfully completed the protocol (89%). There were five complications in the 46 intent-to-treat group (10.9%). CONCLUSIONS: Outpatient craniotomy for brain tumor is a feasible option which appears safe and effective for selected patients. Besides being resource-friendly, the procedure may be psychologically less traumatic to patients than standard craniotomy for brain tumor. Proper prospective studies including satisfaction surveys would help resolve these issues and will be the next step. PMID- 11383934 TI - Improved outcomes in stroke thrombolysis with pre-specified imaging criteria. AB - BACKGROUND: A 1995 National Institute of Neurological Disorders (NINDS) study found benefit for intravenous tissue plasminogen activator (tPA) in acute ischemic stroke (AIS). The symptomatic intracranial hemorrhage (SICH) rate in the NINDS study was 6.4%, which may be deterring some physicians from using this medication. METHODS: Starting December 1, 1998, patients with AIS in London, Ontario were treated according to NINDS criteria with one major exception; those with approximately greater than one-third involvement of the idealized middle cerebral artery (MCA) territory on neuroimaging were excluded from treatment. The method used to estimate involvement of one-third MCA territory involvement bears the acronym ICE and had a median kappa value of 0.80 among five physicians. Outcomes were compared to the NINDS study. RESULTS: Between December 1, 1998 and February 1, 2000, 30 patients were treated. Compared to the NINDS study, more London patients were treated after 90 minutes (p<0.00001) and tended to be older. No SICH was observed. Compared to the treated arm of the NINDS trial, fewer London patients were dead or severely disabled at three months (p=0.04). Compared to the placebo arm of the trial, more patients made a partial recovery at 24 hours (p=0.02), more had normal outcomes (p=0.03) and fewer were dead or severely disabled at three months (p=0.004). CONCLUSIONS: The results of the NINDS study were closely replicated and, in some instances, improved upon in this small series of Canadian patients, despite older are and later treatment. These findings suggest that imaging exclusion criteria may optimize the benefits of tPA. PMID- 11383936 TI - Modified Brooks posterior wiring technique for three-point C1-C2 arthrodesis. AB - BACKGROUND: The optimal surgical treatment of atlanto-axial instability remains controversial despite the variety of modifications and supplemental techniques currently available. METHODS: We describe a modification of the Brooks posterior wiring technique supplemented with transarticular screws for C1-C2 instability. RESULTS: This method has been implemented in 30 patients in our institution with no radiological failures. CONCLUSIONS: The modification provides several technical advantages and potentially stronger fixation compared to methods currently in use. PMID- 11383937 TI - The spectrum of electrophysiological abnormalities in Bell's palsy. AB - BACKGROUND: As part of an investigation of a suspected "outbreak" of Bell's palsy in the Greater Toronto Area, a population-based sample of patients with Bell's palsy was investigated electrophysiologically to help understand the spectrum of abnormalities that can be seen in this setting. METHODS: Two hundred and twenty four patients were surveyed, of whom 91 underwent formal neurological assessment. Of the latter, 44 were studied electrophysiologically using standard techniques. Thirty-two of the 44 patients fulfilled clinical criteria for Bell's palsy. RESULTS: A wide range of electrophysiological changes was observed. Blink responses were the most useful test showing diagnostic sensitivity of 81% and specificity of 94% compared to the contralateral control side. Needle electromyography was additionally helpful in only one patient of six with normal conduction studies. CONCLUSIONS: There is a wide spectrum of electrophysiological abnormalities in Bell's palsy. Blink reflex latencies may be under-utilized in the assessment of the facial nerve in Bell's palsy. Facial EMG is not generally useful in routine assessment. PMID- 11383938 TI - Biogenic amine metabolites and thiamine in cerebrospinal fluid in heredo degenerative ataxias. AB - BACKGROUND: The aims of the present study were: i) to measure levels of the dopamine metabolite homovanillic acid (HVA), the serotonin metabolite 5 hydroxindoleacetic acid (5HIAA) and precursor tryptophan, as well as the noradrenaline metabolite 3-methoxy-4-hydroxyphenylethylene glycol (MHPG) and thiamine in the cerebrospinal fluid (CSF) of patients with Friedreich's ataxia (FA), olivopontocerebellar atrophy (OPCA), and the autosomal recessive spastic ataxia of Charlevoix-Saguenay (ARSAC), as compared with sex- and age-matched control subjects. PATIENTS AND METHODS: CSF amine related compound levels and thiamine results were compared in 40 FA, 44 OPCA and nine ARSAC patients with those of 94 sex- and age-matched subjects. Neuroimaging (CT scans and single photon emission computed tomographies i.e. SPECT) were carried out in all patients and controls. Genetic studies were conducted on OPCA patients. CSF amine related compounds were measured by high performance liquid chromatography, whereas CSF thiamine levels were measured by a microbiological method. RESULTS: FA patients had significantly lower CSF HVA, 5HIAA and thiamine values than control patients and a trend for lower MHPG levels. In OPCA patients, CSF HVA, MHPG and thiamine values were markedly lower whereas CSF 5HIAA values showed only a trend towards lower levels; in ARSAC patients only thiamine and HVA CSF values were lower than those in control subjects. CONCLUSION: After presenting the relationships between neurochemical findings on one side, the degree of ataxia, the degree of cerebellar atrophy and the SPECT findings on the other, the authors concluded that replacement and neuroprotective clinical trials in these patients would have to include two or three drugs because the neurotransmitter deficiencies are multiple. PMID- 11383939 TI - Effect of corticosteroid therapy on serum and CSF malondialdehyde and antioxidant proteins in multiple sclerosis. AB - OBJECTIVE: Multiple sclerosis (MS) is a disease characterised by perivascular infiltrates and demyelination of the white matter in the central nervous system. Although the precise cause of MS remains unknown, some investigations have been carried out on antioxidant mechanisms in these patients. METHODS: In this study, malondialdehyde (MDA), as a lipid peroxidation marker, and ceruloplasmin (Cp) and transferrin (Trf), as antioxidant proteins, levels were determined in cerebrospinal fluid (CSF) and serum of 30 MS patients before and after corticosteroid therapy and in 20 control subjects. Transferrin and Cp levels were measured by the nephelometric method and MDA was measured spectrophotometrically. RESULTS: Mean MDA(serum) and MDA(CSF) levels were found to be highest in the pretreatment group and lowest in the control group. Although there was no significant difference in terms of serum Trf level, serum Cp was found higher in pre- and posttreatment groups than in the control groups. Ceruloplasmin and Trf levels of CSF were not detectable using the nephelometric method. A significant correlation was found between MDA(CSF) and MDA(serum) in the pretreatment group (r=0.58). CONCLUSIONS: These data revealed that lipid peroxidation was increased in serum and particulary in CSF of MS patients and was reduced with corticosteroid therapy. PMID- 11383940 TI - A case-control study of Parkinson's disease in urban population of southern Israel. AB - BACKGROUND: In recent years, an increased prevalence of Parkinson's disease (PD) in southern Israel was observed. The aim of this study was to determine which exposures are associated with PD in the urban population of this region. METHODS: Ninety-three PD patients living in towns were compared to 93 age and sex matched controls. A previously validated questionnaire, including demographic data, education, data on exposures, previous diseases, family history and habits, was administered. RESULTS: In multivariate logistic regression analysis, it was found that history of work in construction sites was the strongest predictor of PD risk, followed by exposure to pesticides. In contrast, there was a negative association with smoking and history of mechanical factory employment. When the same statistical analysis was limited to association of PD with smoking, pesticides and construction work, the latter was found to be the strongest risk factor. CONCLUSION: The risk factors for PD in this population are work on a construction site and exposure to pesticides. PMID- 11383941 TI - Syrinx and tuberculoma formation in tuberculous arachnoiditis. PMID- 11383942 TI - Subacute femoral compressive neuropathy from iliacus compartment hematoma. AB - BACKGROUND: Traumatic retroperitoneal hematoma in the iliacus muscle is an unusual but potentially serious cause of femoral compression neuropathy. CASE REPORT: We describe the clinical, imaging, and management features of a case of traumatic iliacus retroperitoneal hematoma with delayed manifestation of femoral neuropathy. DISCUSSION: The anatomical substrate for hematoma formation with subacute compression of the femoral nerve is emphasized. A subacute compartment syndrome with progressive edema, swelling and ischemia of iliacus compartment is suggested as the underlying cause. Early fasciotomy with or without hematoma evacuation should be considered in order to provide rapid decompression and to minimize the chance of permanent nerve injury. PMID- 11383943 TI - Deep cerebral venous thrombosis: an illustrative case with reversible diencephalic dysfunction. AB - BACKGROUND: Isolated thrombosis of the deep cerebral veins is rare and its diagnosis can be difficult. Mortality is often high and little is known about the long-term prognosis. CASE REPORT: We report a 24-year-old woman with akinetic mutism and extensive bilateral thalamic lesions. CT and MRI allowed early diagnosis by demonstrating thrombosis within the internal cerebral veins, without the need for angiography. Heparin treatment was used safely despite the presence of thalamic and intraventricular hemorrhage. After five weeks, the patient recovered rapidly and remains well at 18 months. Serial MRI showed dramatic resolution of the imaging abnormalities. CONCLUSIONS: The clinical features and characteristic neuroimaging appearance of deep cerebral venous thrombosis should be recognized by physicians caring for stroke patients. Deep cerebral venous thrombosis can produce extensive venous congestion and vasogenic edema without early infarction. Excellent clinical recovery is possible even after severe and prolonged neurological deficits. PMID- 11383944 TI - Sarcoidosis presenting as an intramedullary spinal cord lesion. AB - OBJECTIVES: Sarcoidosis affects the spinal cord in only 0.43% of patients with sarcoidosis. Usually there is systemic involvement prior to the development of cord lesions. We present a case of sarcoid isolated to the intramedullary spinal cord, which was a diagnostic and therapeutic challenge. We review the case and then present a review of the literature with an emphasis on presentation, diagnosis and treatment. METHODS: We have reviewed a patient who presented with an isolated sarcoid granuloma affecting the cervical spinal cord. All pertinent history and physical information was extracted from the patient's chart and through patient interview. Laboratory, radiographic and pathological investigations are presented. RESULTS AND CONCLUSIONS: Fourteen patients have been reported with isolated intramedullary spinal cord sarcoidosis. Current practice supports the role of surgery for biopsy; mainstay of treatment is corticosteroids. PMID- 11383945 TI - Meige syndrome secondary to basal ganglia injury: a potential cause of acute respiratory distress. AB - BACKGROUND: Meige syndrome is a movement disorder that includes blepharospasm and oromandibular dystonias. Its etiology may be idiopathic (primary) or it may arise secondary to focal brain injury. Acute respiratory distress as a feature of such dystonias occurs infrequently. A review of the literature on Meige syndrome and the relationship between dystonias and respiratory compromise is presented. METHODS: A 60-year-old woman suffered a cerebral anoxic event secondary to manual strangulation. She developed progressive blepharospasm combined with oromandibular and cervical dystonias. Neuroimaging demonstrated bilateral damage localized to the globus pallidus. Years later, she presented to the emergency department in intermittent respiratory distress associated with facial and cervical muscle spasms. RESULTS: Increasing frequency and severity of the disorder was noted over years. The acute onset of respiratory involvement required intubation and eventual tracheotomy. A partial therapeutic benefit of tetrabenazine was demonstrated. CONCLUSION: This case highlights two interesting aspects of Meige's syndrome: (1) Focal bilateral basal ganglia lesions appear to be responsible for this patient's movement disorder which is consistent with relative overactivity of the direct pathway from striatum to globus pallidus internal and substantia nigra pars reticularis; (2) Respiratory involvement in a primarily craniofacial dystonia to the point of acute airway compromise. PMID- 11383946 TI - Absent p53 immunohistochemical staining in a pituitary carcinoma. AB - BACKGROUND: Carcinomatous transformation of pituitary adenomas is uncommon, and is generally accompanied by nuclear accumulation of p53 protein. Pituitary carcinoma lacking accumulation of p53 protein is very rare, only two such cases being previously reported. METHODS: A patient presented with visual disturbance and cranial nerve palsies and was found to have a suprasellar mass. He underwent both transphenoidal and transfrontal excision of a nonfunctioning pituitary adenoma which recurred several times. The third recurrence was accompanied by multiple dural-based metastases. Despite aggressive surgical management, he continued to develop additional intracranial lesions and died two years after the discovery of metastatic disease. Specimens from 1984, 1995, 1997 and 1998 were available for histological and immunocytochemical analysis. Antibodies recognizing the pituitary hormones (ACTH, PRL, GH, FSH, LH and TSH), as well as cytokeratin, epithelial membrane antigen (EMA), glial fibrillary acidic protein (GFAP) and chromogranin A were applied to investigate the lineage of the neoplasm. Antisera specific for Ki-67 (MIB-1) and p53 protein were also applied to further delineate the biology of the tumour. RESULTS: Although cytokeratin and chromogranin A were detected in neoplastic cells. no expression of pituitary hormones was demonstrable, indicative of a nonfunctioning, null-cell pituitary adenoma. Nuclear pleomorphism and mitotic activity increased with subsequent resections. Abnormal accumulation of p53 protein was not observed, neither in early resections nor in the metastatic deposits. CONCLUSIONS: Failure to demonstrate p53 protein accumulation does not ensure a favourable outcome for pituitary adenoma. Accordingly, pituitary carcinoma may occur in the absence of p53 accumulation. The factors which underlie aggressive behaviour of pituitary neoplasms are uncertain but are under investigation. PMID- 11383947 TI - Aspects of stroke imaging. PMID- 11383948 TI - Congenital diaphragmatic hernia: update and review. AB - Newborns with congenital diaphragmatic hernia (CDH) present challenges for the neonatal intensive-care nurse. Although CDH has been a known condition for almost 200 years, the treatment strategy for newborns with CDH has changed during the past decade. Despite these improvements, the mortality rate for this condition remains high. An understanding of the anatomical basis and new treatment modalities for this condition will prepare nurses to care for these newborns. PMID- 11383949 TI - The importance of the sexual health history in the primary care setting. AB - Nurses often are apprehensive when inquiring about women's sexual health issues. A comprehensive sexual health assessment, however, is an important part of the health history and interview. Ensuring confidentiality and maintaining professionalism will create the trusting, comfortable environment necessary for a thorough evaluation of a client's sexual health risks. Nurses who are familiar with diverse sexual issues can help women deal with the changes that may occur during the life span. PMID- 11383950 TI - Association between early prenatal care and mother's intention of and desire for the pregnancy. AB - OBJECTIVE: To examine the associations between having planned to become pregnant and valuing an unintended pregnancy with seeking early prenatal care. DESIGN: This historical cohort study, using data from the National Survey of Family Growth (NSFG), Cycle V (1995), examined separately the associations of having a planned pregnancy (timing of pregnancy) and valuing an unintended pregnancy (feelings about the pregnancy) with timing of initiation of prenatal care. PARTICIPANTS: The NSFG was based on a national probability sample of women ages 15 to 44 (N= 1,989). OUTCOME MEASURE: Timing of initiation of prenatal care (early or late/no). Early prenatal care was defined as care sought within the first 13 weeks of pregnancy. It was hypothesized that unwanted and unintended pregnancies were predictors of late or no prenatal care. RESULTS: In agreement with previous research using logistic regression, unwanted pregnancy was found to be a predictor of late/no prenatal care. Black ethnicity also was found to be a predictor of late/no prenatal care. CONCLUSIONS: Interventions that are sensitive to culture and involve patient education regarding family planning are needed. PMID- 11383951 TI - Pregnant women's perceptions of abuse. AB - OBJECTIVE: To determine whether pregnant women's perceptions of abuse severity and danger, and their ability to control the abuse, are significantly correlated with the acts of abuse they experience, and to find out whether relationships exist among women's appraisals of abuse severity, danger, and their perceived ability to stop the abuse. DESIGN: A correlational design was used to compare the abusive acts experienced by the women and their perceptions of that abuse. SETTING: Nine prenatal clinics in urban areas of the northeastern United States. PARTICIPANTS: Sixty-one ethnically diverse, pregnant abused women were interviewed. MAIN OUTCOME MEASURE: Pearson's product-moment correlations were used to examine relationships between abusive experiences and women's perceptions of abuse. RESULTS: Women's perceptions of abuse severity were modestly correlated with threatened (r = .25) and actual violence (r = .36). Perceptions of abuse severity and danger were correlated (r = .41). CONCLUSIONS: Women's beliefs about abuse severity, danger, and their ability to control abuse cannot be fully comprehended by exploring the discrete acts they experience. Further research is needed to identify additional factors that influence those beliefs. PMID- 11383952 TI - Managing the reemergence of anguish: pregnancy after a loss due to anomalies. AB - OBJECTIVE: To explore and describe parents' experiences and feelings when faced with a pregnancy subsequent to a pregnancy in which they had to choose whether or not to terminate following the diagnosis of a fetal abnormality. DESIGN AND METHOD: Exploratory/descriptive study, using the grounded theory method. SETTING: Participants were recruited through physicians in northeastern Florida and throughout the United States via an Internet support network. PARTICIPANTS: Participants included a registered nurse, a physician, and 22 parents (13 women and 9 partners) who were interviewed during a subsequent pregnancy. RESULTS: Mental anguish resulted after a loss due to fetal anomalies. Such parents felt a loss of innocence and a sense of devastation and experienced changes in spousal relationships, fear of hope, isolation, and stigma. This anguish reemerged during a subsequent pregnancy. The parents managed by developing emotional armor, limiting disclosure of the previous and present pregnancy, delaying attachment to the baby, and forming a strong attachment to health care professionals and other people with similar experiences. CONCLUSIONS: Understanding how parents manage the reemergence of mental anguish during a subsequent pregnancy permits health care professionals to assist these parents. Facilitating parents' efforts to develop emotional armor, limit disclosure, delay attachment to the baby, and attach to health care professionals and support groups can mitigate social and psychologic discomfort for these parents. PMID- 11383953 TI - Women's knowledge of Pap smears, risk factors for cervical cancer, and cervical cancer. AB - OBJECTIVE: To review the literature on women's knowledge of Pap smears, risk factors for cervical cancer, and cervical cancer. DATA SOURCES: The review was based on a search of the relevant literature over a 10-year period using MEDLINE and CINAHL. DATA EXTRACTION: Articles from relevant, indexed journals and textbooks published within the past decade were included. Seminal articles were included as appropriate. DATA SYNTHESIS: Risk factors for the development of cervical cancer have been reevaluated. The case for human papillomavirus as the cause of cervical neoplasms has been strengthened. CONCLUSIONS: Cervical cancer is associated with early sexual debut, number of lifetime sexual partners, nonuse of condoms, and infection with human papillomavirus. Cigarette smoking facilitates development of cervical cancer. PMID- 11383954 TI - Vaginal health and infections. AB - Vaginal infections are one of the most common reasons women seek health care. Nurses play an important role not only in the assessment and management of vaginal infections but also in educating women about vaginal health. Understanding the ecology of the vagina, performing a proper microscopy, and recognizing risk factors associated with infections are key to vaginal health. PMID- 11383955 TI - Sexually transmitted viral infections: epidemiology and treatment. AB - Human papillomavirus, herpes simplex virus, and both hepatitis A and B are some of the most common sexually transmitted infections (STI) worldwide. They are preventable, but unlike bacterial STIs the person may harbor the virus in her or his body for life with periodic recurrences of active infection. Viral STIs have long-term health consequences, some of which are serious and life threatening. Nurses not only care for individuals who have a viral STI but also can provide education for prevention. PMID- 11383956 TI - Nonviral sexually transmitted infections. AB - In spite of having standard diagnostic methods and effective treatment regimens, nonviral sexually transmitted infections continue to cause significant morbidity and mortality. Nonviral infections are of special concern in young populations and have more serious consequences in women than in men. With perinatal exposure, newborns are at risk for both minor and major complications, including congenital anomalies, mental impairment, and death. To make an impact on the serious sequelae associated with these infections, nurses must recognize common signs and symptoms, select appropriate diagnostic tests, and rapidly initiate effective treatment. Appropriate emotional support and effective counseling are important components of infection management. PMID- 11383957 TI - The similarities and differences of endometritis and pelvic inflammatory disease. AB - Pelvic inflammatory disease, a common gynecologic problem with an estimated 1 million cases annually, results in serious sequelae. Endometritis, a common obstetric problem occurring in more than 15% of all pregnancies, is the leading cause of maternal mortality. Risk factors are numerous. Diagnosis is not always definitive and relies heavily on the clinical judgment of the practitioner. Prompt initiation of empiric treatment regimens, close monitoring of the client, and client education result in positive health outcomes. PMID- 11383958 TI - Women and HIV/AIDS: act local/think global. AB - The numbers of those affected by the HIV virus worldwide are staggering. Emerging research demonstrates a number of lower cost medical approaches that would significantly reduce the rates of transmission of the virus. Nursing plays a critical role in quality-of-life issues at time of diagnosis. Supportive caregiving by compassionate use of vocabulary and knowledgeable resourcing for patients is part of nursing practice in HIV/AIDS care. PMID- 11383959 TI - Paediatric HIV infection. AB - HIV and AIDS have made a huge global impact, permeating the social, cultural, and economic fabric of almost all nations. The first cases of HIV infection in children were reported in the late 1980s and numbers have since risen steadily throughout the world, with some of the poorest and least developed countries experiencing the highest prevalence. Combined drug regimes have changed the course of HIV-related illness and brought increased survival to those for whom treatment is available. With this, however, have come fresh concerns relating to drug resistance, treatment adherence, and the risk of second-generation vertical transmission as HIV-infected children now survive into adulthood and beyond. The psychological literature has addressed issues such as the direct effect of HIV on child development, social and cultural attitudes, family functioning and support, affected children and orphans of HIV-infected parents, sexual health education, disclosure of diagnosis, and long-term clinical management. The outcome for those living in wealthier countries is optimistic, but the spread of this virus in the rest of the world and its impact on family life and social and political systems remains of great concern. PMID- 11383960 TI - The treatment of bipolar disorder in children and adolescents. AB - This paper reviews the presentation, clinical features. and management of early onset bipolar disorder. A framework for the treatment is based upon a systematic, critical appraisal of the available literature. A multimodal approach is emphasised using individual and family psychoeducational interventions in conjunction with pharmacotherapy. The role of mood stabilisers--lithium--and the anticonvulsants--sodium valproate and carbamazepine is reviewed, alongside the treatments for depression in bipolar disorder and refractory mania. PMID- 11383961 TI - Prevalence of psychiatric diagnoses and the role of perceived impairment: findings from an adolescent community sample. AB - The present study examined psychiatric functioning in a community sample of adolescents aged 14 to 17 years (average age of 15 years). We administered the Diagnostic Interview Schedule for Children-2.25 (DISC-2.25) to 1,201 adolescents and their mothers to obtain prevalence estimates of DSM-III-R disorders and the amount of perceived impairment associated with these disorders. While adolescent females reported a significantly higher prevalence of psychiatric disorders than males (15.5% vs. 8.5%), mothers indicated no sex difference. Compared with adolescent males, females had significantly higher rates of internalizing, anxiety. and depressive disorders. In contrast, the prevalence of externalizing disorders was significantly higher among adolescent males. The inclusion of an impairment criterion had a significant impact in reducing the prevalence rates of overall psychiatric disorders. This reduction occurred mainly through impairment's effects on internalizing disorders, specifically anxiety-based disorders (i.e., simple and social phobia). Given the limited research on the effect of impairment on the prevalence of adolescent psychiatric disorders, future work in this area seems warranted. PMID- 11383962 TI - Modeling interinformant agreement in the absence of a "gold standard". AB - Epidemiological surveys of child and adolescent mental disorders often rely on multiple informants to get a complete diagnostic picture. A consistent finding in the literature is that different informants often do not identify the same children as being disordered. However, because current strategies for estimating interinformant agreement often involve categorizing children using less than perfectly sensitive and/or specific symptoms, biased estimates of interinformant agreement are likely. The aim of this report was to illustrate how latent class analysis (LCA) can be used to model interinformant agreement in the absence of a "gold standard". The proposed model consists of informant-specific latent variables each made up of two or more latent classes corresponding to different levels of symptomatology. Unlike most previous applications of LCA this model allows us to model the extent to which the prevalence of the disorder is the same across informants; and, in addition, the association between informants. The data set comes from a prospective longitudinal study of 2,264 children from Quebec (1,155 boys and 1,109 girls). In grade 2, teachers and mothers independently rated each child on three physical aggression behavior symptoms. We satisfactorily accounted for the cross-classification of the behavior symptoms by postulating the existence of two latent variables--one for each informant each made up of three latent classes of children: low-, medium-, and high-aggressive. The results showed that the prevalence of low- and medium-aggressive children in the population differed from teacher to mother, but that the prevalence of high aggressive children did not. We found that the association between teacher and mother was large and positive and did not vary according to the child's physical aggression state or gender; in contrast, the association between physical aggression and gender was not the same for mother and teacher. Limitations and other potential applications of the proposed model are discussed. PMID- 11383963 TI - Comparison of multiple measures of ADHD symptomatology: a multivariate genetic analysis. AB - The phenotypic and genetic interrelationships underlying ADHD symptomatology assessed by various instruments were examined on a sample of 735 male and 819 female same-sex twin pairs, aged 8 to 16 years, participating in the first phase of the Virginia Twin Study of Adolescent Behavioral Development (VTSABD). Multivariate analyses were applied to parental and teacher ratings from an investigator-based interview, the CAPA, and three questionnaires (the CBCL and the Rutter Parent and Teacher Scales). Results from patterns of intercorrelations and factor analyses of maternal measures suggested that at the phenotypic level, these assessed the same underlying behavioural construct, which differed from other emotional and behavioural constructs. However, genetic analyses showed that in addition to a common factor underlying the expression of ADHD as assessed across the range of measures, additional genetic factors were identified that were method- and rater-specific. The findings suggest that although the investigator-based interview and the behavioural checklists tap similar aspects of ADHD behaviour, there is additional rater-specific variance. PMID- 11383964 TI - The comorbidity of ADHD in the general population of Swedish school-age children. AB - This study examined patterns of comorbid/associated diagnoses and associated problems in a population sample of children with and without DSM-III-R attention deficit hyperactivity disorder (ADHD). Half (N = 409) of a mainstream school population of Swedish 7-year-olds were clinically examined, and parents and teachers were interviewed and completed questionnaires. The children were followed up 2-4 years later. Eighty-seven per cent of children meeting full criteria for ADHD (N = 15) had one or more and 67% at least two--comorbid diagnoses. The most common comorbidities were oppositional defiant disorder and developmental coordination disorder. Children with subthreshold ADHD (N = 42) also had very high rates of comorbid diagnoses (71% and 36%), whereas those without ADHD (N = 352) had much lower rates (17% and 3%). The rate of associated school adjustment, learning, and behaviour problems at follow-up was very high in the ADHD groups. We concluded that pure ADHD is rare even in a general population sample. Thus, studies reporting on ADHD cases without comorbidity probably refer to highly atypical samples. By and large, such studies cannot inform rational clinical decisions. PMID- 11383965 TI - Which aspects of ADHD are associated with tobacco use in early adolescence? AB - Several studies have found a relationship between attention-deficit hyperactivity disorder (ADHD) and substance use, primarily in the context of co-occurring conduct disorder (CD). However, very few have examined the associations between the individual dimensions of ADHD (hyperactivity-impulsivity and inattention) and substance use, even though these dimensions reflect distinct symptom groupings, both by clinical definition (DSM-IV, American Psychiatric Association, 1994) and through empirical demonstration (Lahey et al., 1988: McBurnett et al., 1999). This longitudinal study examines the relationship between dimensions of ADHD (as described by DSM) and substance use, accounting for other psychopathology and factors potentially related to substance use. Participants were 177 clinic referred boys (initially between ages 7 and 12) followed up over nine annual phases until all participants had reached age 15. Annual assessment included structured clinical interviews with parent and child and self-report questionnaires of substance use, as well as questionnaires related to family factors and parenting behaviors. Seventy-eight per cent of participants reported use of tobacco, alcohol, marijuana, or other illicit drugs during adolescence, with 51% reporting any tobacco use. The inclusion of CD rendered all bivariate relationships with the full diagnosis of ADHD nonsignificant. However, adolescent inattention, considered independently, was associated with a 2.2 times greater risk for concurrent tobacco use, even after controlling for CD. Even when other factors, selected based on their associations with tobacco use in adolescence, were included in a regression model (concurrent adolescent CD odds ratio [OR] = 6.08), duration of tobacco use by age 12 (OR = 5.11), poor parental communication in childhood (OR = 2.9), African-American ethnicity (inversely predictive; OR = 0.15), inattention (OR = 2.3) remained significantly associated with tobacco use in early adolescence. These findings highlight the importance of considering the risks for comorbid substance use separately by individual dimensions of ADHD. PMID- 11383966 TI - Developmental trajectories of physical aggression from school entry to late adolescence. AB - The developmental perspective applied to psychopathology has led to the concept of early- and late-onset disorders. This study explores the application of the early- and late-onset concepts of antisocial behavior to physical aggression. Are there two categories of chronically physically violent adolescents: those who are physically aggressive throughout childhood and those who start being physically aggressive during adolescence? The estimation of developmental trajectories for repeated measures of two different response variables physical aggression in childhood as measured by teacher reports and physical aggression in adolescence as measured by self-reported violent delinquency is achieved with a semi parametric, group-based method. This new method is applied to a large sample of males from Montreal who have been assessed repeatedly since kindergarten. Several salient findings emerge from the analysis. First, we find considerable change in the levels of childhood and adolescent physical aggression. Thus, there is little evidence of stability of behavior in an absolute sense. A second key finding concerns the connection of childhood aggression to adolescent aggression. Boys with higher childhood physical aggression trajectories are far more likely to transition to a higher-level adolescent aggression trajectory than boys from lower childhood physical aggression trajectories. However, for all childhood physical aggression trajectory levels the modal transition is to a relatively low level adolescent aggression trajectory. Third, we find little evidence of "late onset" of high-level physical aggression. Specifically, the joint trajectory analysis finds no evidence of transition from a low physical aggression trajectory in childhood to a high trajectory in adolescence. PMID- 11383967 TI - Perceived maternal control and support: effects on hostile biased social information processing and aggression among clinic-referred children with high aggression. AB - Based on a sample of 89 aggressive clinic-referred children, aged between 9 and 11 years, a longitudinal study of 1-year duration was conducted to examine (a) whether the children's perception of control and support of their mothers' to them predicted their hostile attribution of intent and hostile response selection, and (b) whether these hostile biased social cognitions mediated the relationships between their perceived maternal behaviours and their aggression. Participants completed a questionnaire covering both the perceived maternal control and support. One year later, their hostile attribution of intent and response selection, and aggression, were measured. Results showed that perceived maternal control and perceived maternal support were associated positively and negatively, respectively, with both the social cognition measures. Also, the social cognition measures mediated the relationships of the perceived maternal measures with aggression. The findings are discussed in terms of how children's hostile biased relational schemas and scripts, developed from negative parenting and insecure attachment, favour more hostile social cognitions, and how these in turn mediate children's current hostile biased social behaviours. PMID- 11383968 TI - Do high-threat life events really provoke the onset of psychiatric disorder in children? AB - Studies on adults have suggested important effects of stressful life events in provoking onset of psychiatric disorder. Only a few comparable studies on children exist, and their results are inconsistent in relation to definite timing effects. Meeting some important methodological challenges overlooked in the past research, this study set out to examine whether the onset of psychiatric disorder in children was more likely to occur shortly after a severe event, as compared with other times. The sample consisted of 99 consecutive, newly referred patients, aged 8-16 years, from a child psychiatry service in London. PACE (Psychosocial Assessment of Childhood Experiences), an investigator-based, standardized interview was used to assess the timing and impact of life events over the preceding 18 months. CAPA (Child and Adolescent Psychiatric Assessment), a standardized diagnostic assessment, was used to establish the presence, timing, and consequential impairment of child and adolescent psychiatric symptoms. In a within-subject, over-time design, conditional logistic regression techniques were employed to examine whether risk of onset was greater in the 9 weeks following a high-threat life event than at other times. There was a small but statistically significant association between child-reported events and child-reported onset; the associations with parent-reported onset were inconsistent. Parent-reported events failed to relate to onset by either source. The study offers only quite limited support to the notion of negative life events provoking onset of psychiatric disorder in children and young people. The possible reasons for this are discussed, together with important conceptual and methodological issues to problems of defining onset, and the choice of appropriate designs for data analysis. PMID- 11383969 TI - Home is where it begins: parents, children, and stressful events. AB - A 20-event, 7-point scale on the stressfulness of selected unpleasant experiences was administered to 364 children in Grades 4 to 6 in an American intermediate school. Independently, a parallel questionnaire was given to their parents to indicate (a) how upsetting they would find each event themselves, (b) what they estimate their own children's ratings to be, and (c) whether they think their children have actually experienced each event. The parent-child agreements among the 239 responding pairs were quite close on (1) the scale values (the median upsettingness ratings) with an r of .96, (2) the discriminal dispersions (the interquartile ranges of ratings) with an r of .79, and (3) the incidences with an r of .96. The striking contrasts with the earlier comparisons with professional judgements invite a reassessment of parents as dependable observer-reporters. PMID- 11383970 TI - Susceptibility to illusions and performance on visuospatial tasks in individuals with autism. AB - The current study follows a recent paper reporting that individuals with autism were just as susceptible to visual illusions as those without autism (Ropar & Mitchell, 1999). The possibility that individual differences may account for the failure to replicate Happe's (1996) findings is explored by presenting a battery of visuospatial tasks thought to measure weak central coherence (embedded figures, block design, Rey complex figure test). Participants with autism were distinguished by relatively good performance on visuospatial tasks, though there was no superiority effect in those with Asperger's syndrome. Performance on the visuospatial battery did not significantly predict susceptibility to illusions in various participant groups, including those with autism and Asperger's syndrome. This suggests that perception of illusions and performance on visuospatial tasks may rely on different mechaisms. The implications for the theory of weak central coherence are discussed. PMID- 11383971 TI - Individual differences in cognitive planning on the Tower of Hanoi task: neuropsychological maturity or measurement error? AB - The Tower of Hanoi (ToH) task was given to 238 children aged from 7 to 15 years, and 20 adults. Individual variation within an age band was substantial. ToH score did not correlate significantly with Verbal IQ, nor with ability to inhibit a prepotent response. We readministered the ToH to 45 children after 30 to 40 days. The test-retest correlation of .5 is low in relation to accepted psychometric standards, though at least as high as reliability of the related Tower of London (ToL) in adults. The reasons for low reliability remain unclear: task novelty did not seem to be involved, as children did not improve on retest. We conclude that it is not safe to use this test to index integrity or maturation of underlying neurological systems in children. We compared our results with three published studies using the ToL with children, and found similar levels of performance on problems involving the same number of moves. Another study using automated ToL obtained much poorer scores, suggesting that computerised presentation may impair children's performance. PMID- 11383972 TI - Research into the specificity of depression after stroke: a review on an unresolved issue. AB - Iwo decades of research have failed to generate consistent insight into the specificity of poststroke depression (PSD). This is, at least in part, caused by methodological difficulties. Differences in symptom profile between PSD and depression with no or another medical cause were described, but no specific and unequivocal clinical picture has been established so far. Prevalence rates of PSD varied largely between studies. In community based studies using standardised diagnostic instruments for depression, relatively low prevalence rates were reported compared to inpatient or rehabilitation studies. PSD occurs most frequently in the first few months after stroke, while a new incidence peak may occur 2-3 years after stroke. Two systematic reviews on the relation between lesion location and depression did not support the claim that left hemisphere lesions are a risk factor for PSD. A new concept of vascular depression has been proposed, which relates depression in the elderly to acute or chronic damage to the cerebral vascular system. Future efforts should aim at increasing the uniformity of study designs, assessment tools should be further improved for use in cognitively impaired patients and appropriate control groups should be defined to study the characteristic features of PSD. PMID- 11383973 TI - Schizophrenia, a neurodegenerative disorder with neurodevelopmental antecedents. AB - Schizophrenia is a devastating disorder that has been referred to as youth's greatest disabler. Although a number of hypotheses have been proposed in an attempt to explain the pathophysiology of schizophrenia no single theory seems to account for all facets of the disease. Each hypothesis explains some of the phenomena associated with schizophrenia and it is probable that many variables described in these hypotheses interact to produce a disorder characterized by heterogeneous symptomatology, progression and prognosis. Compelling evidence suggests that the primary disturbance is a neurodevelopmental abnormality, possibly resulting from a genetic defect(s), resulting in a predisposition to schizophrenia. Events later in life may then lead to the presentation of symptoms and a subsequent progression of the disease. Recent evidence suggests that the progressive course of schizophrenia is associated with ongoing neurodegenerative processes. Changes in brain derived neurotrophic factor (BDNF) may explain the various changes observed in schizophrenia. PMID- 11383974 TI - Pharmacotherapy of mental illness--a historical analysis. AB - The history of pharmacotherapy of mental illness can be divided into three periods. Introduction of morphine, potassium bromide, chloral hydrate, hyoscine, paraldehyde, etc., during the second half of the 19th century (first period), led to the replacement of physical restraint by pharmacological means in behavior control. Introduction of nicotinic acid, penicillin, thiamine, etc., during the first half of the 20th century (second period), led to significant changes in the diagnostic distribution of psychiatric patients; psychoses due to cerebral pellagra, and dementia due to syphilitic general paralysis virtually disappeared from psychiatric hospitals, and the prevalence of dysmnesias markedly decreased. Treatment with therapeutically effective drugs of mania, schizophrenia, depression, bipolar disorder, generalized anxiety disorder, panic disorder, obsessive compulsive disorder, Alzheimer's disease, etc., during the second half of the 20th century (third period), brought to attention the heterogeneity of the populations within the diagnostic categories of schizophrenia and depression. Introduction of the first set of psychotropics and the spectrophotofluorimeter during the 1950s triggered the development of neuropsychopharmacology. Introduction of genetic technology for the separation of receptor subtypes in the 1980s opened the path for the "tailoring" of psychotropic drugs by the dawn of the 21st century, to receptor affinities. PMID- 11383975 TI - Role of endogenous opioid system in the regulation of the stress response. AB - Numerous studies and reviews support an important contribution of endogenous opioid peptide systems in the mediation, modulation, and regulation of stress responses including endocrine (hypothalamopituitary-adrenal, HPA axis), autonomic nervous system (ANS axis), and behavioral responses. Although several discrepancies exist, the most consistent finding among such studies using different species and stressors is that opioids not only diminish stress-induced neuroendocrine and autonomic responses, but also stimulate these effector systems in the non-stressed state. A distinctive feature of the analgesic action of opioids is the blunting of the distressing, affective component of pain without dulling the sensation itself. Therefore, opioid peptides may diminish the impact of stress by attenuating an array of physiologic responses including emotional and affective states. The widespread distribution of enkephalin (ENK) throughout the limbic system (including the extended amygdala, cingulate cortex, entorhinal cortex, septum, hippocampus, and the hypothalamus) is consistent with a direct role in the modulation the stress responses. The predictability of stressful events reduces the impact of a wide range of stressors and ENK appears to play an important role in this process. Therefore, ENK and its receptors could represent a major modulatory system in the adaptation of an organism to stress, balancing the response that the stressor places on the central stress system with the potentially detrimental effects that a sustained stress may produce. Chronic neurogenic stressors will induce changes in specific components of the stress induced ENKergic system, including ENK, delta- and mu-opioid receptors. This review presents evidences for adaptive cellular mechanisms underlying the response of the central stress system when assaulted by repeated psychogenic stress, and the involvement of ENK in these processes. PMID- 11383976 TI - The role of cannabinoids in neurodegenerative diseases. AB - An understanding of the actions of Cannabis (Marijuana) has evolved from folklore to science over the previous hundred years. This progression was spurred by the discovery of an endogenous cannabinoid system consisting of two receptors and two endogenous ligands. This system appears to be intricately involved in normal physiology, specifically in the control of movement, formation of memories and appetite control. As we are developing an increased understanding of the physiological role of endocannabinoids it is becoming clear that they may be involved in the pathology of several neurological diseases. Furthermore an array of potential therapeutic targets is being determined--including specific cannabinoid agonists and antagonists as well as compounds that interrupt the synthesis, uptake or metabolism of the endocannabinoids. This article reviews the recent progress in understanding the contribution of endocannabinoids to the pathology and therapy of Huntington's disease. Parkinson's disease, schizophrenia and tremor. PMID- 11383977 TI - The immune system, depression and the action of antidepressants. AB - It is well established that the hypothalamic-pituitary-adrenal axis (HPA) is activated by both external and internal stressors which result in the hypersecretion of adrenal glucocorticoids. In major depression the prolonged elevation of the glucocorticoid concentration leads to a desensitisation of the central glucocorticoid receptors and probably those receptors located on macrophages. These changes may account for the observation that many aspects of cellular immunity are activated in depression (for example, the increased release of pro-inflammatory cytokines from activated macrophages in the periphery and brain, and the increased release of acute phase proteins from the liver) even though other aspects of immunity (for example, natural killer cell activity and T cell replication) are depressed. It is also known that some of the pro inflammatory cytokines are potent activators of the HPA axis. Evidence is provided that the consequences of the hypersecretion of glucocorticoids and pro inflammatory cytokines result in the malfunctioning of noradrenergic and serotonergic neurotransmission in the brain, changes which are reflected in the major symptoms of depression. Support for this view is provided by observations of the effects of some of these cytokines in non-depressed individuals being treated with pro-inflammatory and related cytokines for cancer. This has led to the hypothesis that depression is a form of sickness behaviour which forms the basis of the macrophage theory of depression. The review concludes with a discussion of the role of antidepressants in attenuating the adverse effects of glucocorticoids and pro-inflammatory cytokines on central neurotransmission. Although the precise mechanisms whereby antidepressants these changes is uncertain, there is evidence that they reduce the release of pro-inflammatory cytokines from activated macrophages and thereby facilitate the feedback inhibition of the HPA axis; this results in a reduction in the release of glucocorticoids from the adrenal glands. In addition, many antidepressants have been shown to increase the release of endogenous cytokine antagonists such as interleukin-1 receptor antagonist and interleukin-10. Evidence is also presented to show that different classes of antidepressants act as cyclooxygenase inhibitors which, by lowering the concentration of inflammatory prostaglandins in the brain, reduce the detrimental impact of the inflammatory changes on neurotransmitter function. An advantage of the macrophage hypothesis is that it extends the biogenic amine hypothesis of depression to take account of changes in the endocrine and immune systems which also play a crucial role in the aetiology of depression. In addition, the macrophage hypothesis may broaden the basis of understanding the mechanism of action of antidepressants. PMID- 11383978 TI - The role of the brain reward system in depression. AB - The goal of this review is to familiarize the reader about the potential involvement of the brain reward system (BRS) in symptoms of Major Depressive Disorder (MDD). The authors introduce a novel approach to study the pathophysiology of MDD that includes pharmacological probing of BRS pathways (e.g. d-amphetamine, hydromorphone) together with an elicited and measurable behavioral component (e.g. pleasant effects, increased energy, altered cognition). To this date, the major focus of MDD pathophysiology studies has been to characterize biological differences between healthy subjects and depressed patients such as alteration in the monoaminergic and endocrine systems. The relative importance of the various biological changes has not been elucidated, that is, linking these with specific behavioral manifestations in MDD have rarely been attempted. One core symptom of MDD is a decreased experience of pleasure or interest in previously enjoyed activities (i.e. anhedonia) such as work or hobbies, and is accompanied by decreased motivation or drive. The BRS consists of the neural pathways involved in eliciting rewarding experiences in animals and humans. The hypothesis is that altered BRS function may be an underlying brain mechanism of the loss of pleasure/interest experienced in MDD, and will be manifested through an altered response to a BRS probe. The authors have examined BRS function in MDD by introducing a pharmacological probe (i.e. d-amphetamine/d amph). Amphetamine is defined as a probe due to its ability to release dopamine within major components of the BRS (i.e. the mesocorticolimbic dopamine system.) In addition to the objective pharmacological effects (e.g. altered heart rate), BRS probes like d-amph elicit reliable and measurable behavior, that is, the hedonic effects. A review of the neurobiology of MDD, the BRS, the rationale for implicating the BRS in depressive symptoms, and preliminary data, are presented in this article. PMID- 11383979 TI - The effect of single oral doses of zopiclone on nocturnal melatonin secretion in healthy male volunteers. AB - The effects of single oral doses of zopiclone and temazepam were investigated in eight healthy male volunteers using a single blind, placebo controlled cross over study. Doses of zopiclone were 7.5 and 15 mg while the dose of temazepam was 20 mg. Each dose was separated by at least a one-week washout period. For each subject the dim light melatonin onset (DLMO) was determined on a screening night and the drugs were administered at the time of the DLMO. Melatonin concentrations were determined by radioimmunoassay from plasma samples collected throughout the night. Both temazepam and zopiclone tended to reduce the amount of melatonin secreted, as determined by the area under the plasma concentration time curve. The differences from placebo were not statistically significant (F 3.31 = 1.07, P > 0.1). Similarly a repeated measures analysis of variance on the plasma concentration-time curves did not show any statistically significant differences between drugs and placebo (F 3.28 = 1.15, P > 0.1). There was no evidence from this study of a phase shifting effect of the drugs used. The reasons for the lack of effect on melatonin may be due to the differences in potency of the interaction of these drugs with the GABA-benzodiazepine-chloride ion channel. PMID- 11383980 TI - Function of the hippocampus in memory formation: desperately seeking resolution. AB - Despite considerable efforts and successes investigating the function of the hippocampal formation in memory processes, there are still numerous elusive key issues. Some of them will be addressed in this review. We will argue that recent evidence supports hippocampal participation in several memory processes, such as encoding, short-term and long-term consolidation and retrieval. While some processes, for example encoding and short-term consolidation, have been the subject of detailed investigations, at least for specific and repeatedly used behavioural paradigms, there appears to be considerable lack of information with respect to other processes, for example long-term consolidation. Although the existence of long-term consolidation is not at debate here, there is only very fragmented information as to the cellular processes enabling long-term consolidation. Recent ample evidence now suggests a potential role in metabotropic glutamate receptors, and more specifically the phospholipase C coupled receptor 5, in long-term consolidation. The hyperexpression of receptor protein was limited to CA1 indicating a specific role of this brain region in the consolidation of memories. Future work should further explore this important issue especially since long-term consolidation appears to be a necessity for permanent storage of information, and may thus engage memory mechanism that fail during ageing and dementia. PMID- 11383981 TI - The mechanism of lithium action: state of the art, ten years later. AB - Lithium is an effective drug for both treatment and prophylaxis of bipolar disorder. However, the mechanism of lithium action is still unknown. The inositol depletion hypothesis is supported by biochemical and behavioral data in rats, but primate inositol levels are higher than in rodents and may obviate the effects of depletion. Inhibition of 5HT autoreceptors by lithium is supported by biochemical and behavioral data in rats but would seem more related to lithium's antidepressant than to its antimanic or prophylactic effects. Lithium induces increases in levels of the anti-apoptotic factor Bcl-2. This effect could be most relevant for treatment of neurodegenerative disorders. Lithium inhibits glycogen synthase kinase-3, which is involved in a wide range of signal transduction pathways. However, this lithium effect occurs at high concentrations and may be more relevant for its toxic effect. Lithium in low concentrations induces accumulation of PAP, which affects several cellular processes including RNA processing. However, PAP phosphatase is present more in peripheral tissues than in brain. This lithium effect could explain some of its peripheral side effects. Chronic lithium administration upregulates glutamate reuptake and thus decreases glutamate availability in synapse. Glutamate is an excitatory neurotransmitter and its reduction could exert an antimanic effect. Biochemical and clinical experiments are necessary to determine the key mechanism of lithium efficacy in treatment and prophylaxis of affective disorders. PMID- 11383982 TI - Abnormal neurochemical asymmetry in the temporal lobe of schizophrenia. AB - Neuroanatomical asymmetries are known to be present in the human brain, and loss of reversal of these asymmetries, particularly through changes in the left temporal lobe, have been found in the brains of patients with schizophrenia. In addition to disturbed neuroanatomical asymmetries, disturbed neurochemical asymmetries have also been reported in the brains of patients with schizophrenia. However, in the temporal lobe, the laterality of most of these neurochemical changes has not been specifically evaluated. Few neurochemical studies have addressed left-right differences in the superior temporal gyrus (STG). A deteriorated serotonin2A receptor-G protein qalpha (Gqalpha)-phosphoinositide specific phospholipase C beta1(PLC beta1) cascade has been found in the left, but not right, STG of patients with schizophrenia. Not only neuroanatomical but also neurochemical evidence supports the loss or reversal of normal asymmetry of the temporal lobe in schizophrenia, which might be due to a disruption of the neurodevelopmental processes involved in hemispheric lateralization. PMID- 11383983 TI - Depression and myocardial infarction: relationship between heart and mind. AB - There is a relationship between depression and Myocardial Infarction (MI) as higher levels of depression and severe depression (major vs minor) are associated with higher morbidity and mortality due to cardiac events, which are mainly caused by arrhythmia. Second, severity of MI is not or even inversely related to development of depression. Depression post-MI goes often unrecognized as only 10% of depressed MI patients are diagnosed as such. This underestimation of depression is attributed to its atypical profile, tendency of physicians to interpret depressive symptoms as a transient and 'natural' reaction to a life threatening event, and the scarce knowledge of risk factors associated with development of post-MI depression. During the first 18 months following MI major depression occurs in 15-30% of patients. Depression should be assessed in an early stage as depression has the highest prevalence in hospital and in the first 6 months post-MI. Risk factors for developing post-MI depression include complications during hospitalization, prescription of benzodiazepines during hospitalisation, previous history of depression, and not being able to stop smoking. Selective Serotonin Reuptake Inhibitors (SSRIs) appear to be first choice treatment in post-MI depression. As yet there is no information on the efficacy and safety of Serotonin and Noradrenalin Reuptake Inhibitors (SNRIs). PMID- 11383984 TI - Anxiety/aggression--driven depression. A paradigm of functionalization and verticalization of psychiatric diagnosis. AB - A new subtype of depression is proposed, named: anxiety/aggression-driven depression. The psychopathological, psychopharmacological and biochemical evidence on which this construct is based, is being discussed. Selective postsynaptic 5-HT1A agonists together with CRH and/or cortisol antagonists are hypothesized to be a specific biological treatment for this depression type, in conjunction with psychological interventions to raise the stressor-threshold and to increase coping skills. The development of this depression construct has been contingent on the introduction of two new diagnostic procedures, called functionalization and verticalization of psychiatric diagnosis. These procedures are explained and it is stressed that they are essential to psychiatric diagnosing, in order to put this process on a scientific footing. PMID- 11383985 TI - The psychopharmacology-molecular biology interface: exploring the behavioural roles of dopamine receptor subtypes using targeted gene deletion ('knockout'). AB - In the absence of selective agonists and antagonists able to discriminate between individual members of the D1-like and D2-like families of dopamine receptor subtypes, functional parcellation has remained problematic. 'Knockout' of these subtypes by targeted gene deletion offers a new approach to evaluating their roles in the regulation of behaviour. Like any new technique, 'knockout' has associated with it a number of methodological limitations that are now being addressed in a systematic manner. Studies on the phenotype of D1(A/1), D(1B/5), D2, D3 and D4 'knockouts' at the level of spontaneous and agonist/antagonist induced behaviour are reviewed, in terms of methodological issues, neuronal implications and potential clinical relevance. Dopamine receptor subtype 'knockout' is a nascent technology that is now beginning to fulfil its potential. It is being complemented by more systematic phenotypic characterisation at the level of behaviour and additional, molecular biologically-based approaches. PMID- 11383986 TI - In situ anterior vaginal wall sling. AB - PURPOSE: The aim of this article is to review the indications, technique, and outcomes of the in situ anterior vaginal wall sling (AVWS) for treatment of female stress urinary incontinence. MATERIALS AND METHODS: The operative techniques and the published literature on outcomes of the AVWS were reviewed. RESULTS: Success rates in the range from 70% to 95% were noted. Some authors found that patients with a lower Valsalva leak point pressure (VLPP) had lower success rates than those with a higher VLPP. CONCLUSIONS: The AVWS is a technically straightforward minimally invasive procedure for treatment of stress urinary incontinence that minimizes patient morbidity and avoids some of the potential problems associated with other sling materials. PMID- 11383987 TI - Inguinovaginal sling procedure for female stress incontinence: introduction of a minimally invasive technique. AB - PURPOSE: The inguinovaginal sling procedure is well accepted for surgical treatment of female stress urinary incontinence. Although the functional results are excellent, the operative trauma is higher compared to that of more recently used minimally invasive techniques. MATERIALS AND METHODS: A modified inguinovaginal sling procedure was performed in 15 patients with urodynamically diagnosed intrinsic sphincter deficiency. With the assistance of a laparoscope, two fascial strips were dissected using two small suprapubic skin incisions. The pullthrough maneuver of the fascial slings was facilitated by opening the perivesical space with an inflatable balloon. RESULTS: There were no intraoperative or postoperative complications. Mean follow-up of 7.7 months (range 4 to 11) was available in all patients. Stress incontinence was cured in 14 patients; only one woman reported using one pad per day. All patients were able to void spontaneously after a mean of 8 days (range 6 to 10). CONCLUSIONS: The laparoscopically assisted inguinovaginal sling procedure is less invasive than the original technique. Good long-term results are known from the original inguinovaginal sling procedure, and there is no need for allografts or synthetic material. PMID- 11383988 TI - Delivery of injectable agents for treatment of stress urinary incontinence in women: evolving techniques. AB - PURPOSE: Periurethral bulking agents continue to be used as a minimally invasive alternative for management of stress urinary incontinence in men and women. Agents and delivery techniques will be evaluated and compared. MATERIALS AND METHODS: The only agents currently approved by the United States Food and Drug Administration (FDA) include glutaraldehyde cross-linked collagen, autologous fat, and carbon bead/carrier gel technology. Several other agents are undergoing FDA trial evaluation. These agents differ in material characteristics (particle size, viscosity) that, in some cases, produce different immediate and delayed tissue responses or reactions. RESULTS: Initial subjective cure rates with collagen are acceptable, but the majority of women require reinjection. The risk of allergic phenomena complicates collagen use. Carbon-based material appears to parallel collagen in durability, with the significant advantage of a nonimmunogenic response within host tissues. Autologous fat injection achieves early results similar to those with collagen, but is limited by resorption and fibrous replacement as well as local discomfort associated with harvesting procedures. Experience with newer agents is limited. Patient characteristics also influence response to injectable agents. CONCLUSIONS: Injectable agent materials and delivery techniques continue to evolve. The optimal material is yet to be defined. PMID- 11383989 TI - Treatment options for outlet obstruction following anti-incontinence surgery in females. AB - Urethral obstruction following surgical treatment of stress incontinence can result in a wide spectrum of symptoms ranging from mild urgency to complete retention at its most extreme. Management of these symptoms can be challenging and requires a careful history, pelvic examination, and urodynamics in some instances. In patients refractory to medications or conservative measures, urethrolysis is commonly used to treat the underlying outlet obstruction. The selection of which urethrolysis technique to use can be difficult, as many different methods have been described. In many cases, the choice of technique can be tailored to the original procedure(s) that caused the obstruction. Multiple urethrolysis procedures using a transvaginal approach have been described. One type involves perforation of the endopelvic fascia through a lateral approach. This urethrolysis usually is successful in treating obstruction after needle suspensions and pubovaginal slings, whereas retropubic procedures such as a Marshall-Marchetti-Krantz (MMK) or Burch procedure may require an anterior urethrolysis using a suprameatal approach. In cases where a Burch or MMK was performed and a sling subsequently placed, a combined lateral and anterior approach may be needed to treat the obstruction. In certain cases, a suprapubic approach may be required when the vaginal anatomy was drastically altered or a prior transvaginal urethrolysis failed. Using a urethrolysis technique tailored to the procedure that caused the obstruction, most cases of iatrogenic urethral obstruction should be treatable and result in resumption of normal voiding without producing stress urinary incontinence. PMID- 11383990 TI - Surgical procedures for the treatment of urge incontinence. AB - PURPOSE: Cure of urge incontinence refractory to conservative management may require the application of several surgical techniques. The Ingelman-Sundberg bladder denervation procedure, detrusor myectomy, and augmentation cystoplasty are among the surgical possibilities aimed at dealing with severe urgency and uninhibited detrusor contractions. METHODS AND MATERIALS: A review of the literature was performed to evaluate surgical treatments for refractory urgency and urge incontinence with a focus on the Ingelman-Sundberg bladder denervation procedure, detrusor myectomy, and augmentation cystoplasty. RESULTS: The Ingelman Sundberg bladder denervation has a complete response rate of 54% at a mean of 44.1 months. Among a heterogeneous group of patients, the detrusor myectomy resulted in improvement of compliance and/or resolution of uninhibited detrusor contraction in 63%. The augmentation cystoplasty has the highest overall rate of success but with a much higher likelihood of early and late postoperative complications. CONCLUSIONS: Surgical procedures for urge incontinence have a reasonable success rate with respect to cure of symptoms and urodynamic improvement when present. The possibility of cure or improvement appears to vary directly with the invasiveness of the procedure. However, a logical progression from least to most invasive should be undertaken unless very poor compliance and upper tract abnormalities dictate a more aggressive initial course of action. PMID- 11383991 TI - Fast magnetic resonance imaging of pelvic organ prolapse. AB - Pelvic organ prolapse is abnormal displacement of the pelvic organs from their normal anatomical position. Patients may present with a variety of symptoms, including pain, incontinence, constipation, urinary retention, and defecatory dysfunction. Any combination of cystocele, rectocele, enterocele, sigmoidocele, peritoneocele, and prolapse of the vagina and uterus may occur. Therefore, accurate preoperative evaluation of each organ is important for proper surgical planning. Compared with physical examination and other imaging modalities, advantages of magnetic resonance imaging (MRI) include a global multiplanar view of the pelvis, and the lack of ionizing radiation and invasive procedures. Subsecond MRI techniques have not only shortened the imaging time to minimize motion artifacts but provide the capability for dynamic MRI. In this pictorial essay, we describe fast MRI techniques, MRI findings, and the associated clinical findings in patients with pelvic organ prolapse. We also refer to limitations of MRI. PMID- 11383992 TI - Transabdominal approach to repair of vaginal vault prolapse. AB - PURPOSE: To review the preoperative evaluation of women with vaginal vault prolapse and describe the surgical methods of treatment using a transabdominal approach. METHODS: Abdominal sacral colpopexy is the most widely performed method of transabdominal correction of vaginal vault prolapse. The procedure is completed by securing the apex to the vagina to the periosteum of the sacrum with mesh. This procedure is demonstrated in great detail. Successful repair can be achieved by other transabdominal approaches and by laparoscopic approaches. RESULTS: Twenty women (mean age 67.9 years) were evaluated for complex pelvic floor prolapse. Six (30.0%) patients had failed transvaginal sacrospinus ligament fixation. Abdominal sacrocolpopexy utilizing Marlex mesh, Halban culdeplasty, and paravaginal repair was performed on all patients. Five posterior repairs and one anterior repair was done. The average operating time for the colpopexy and enterocele repair alone is approximately 90 minutes. The average blood loss was 284 cc. The average hospital stay was 3.7 days. The mean follow-up is 11.3 months (6-27 months). The vaginal vault is well supported in all patients with no recurrent enterocele or vault prolapse. Three patients have asymptomatic grade II cystoceles, and three patients have asymptomatic grade II rectoceles. There were few complications. No mesh complications have been encountered. CONCLUSIONS: Vaginal vault prolapse can be a difficult problem to diagnose and treat. Successful treatment requires thorough knowledge of the anatomy, methods of diagnosis, and treatment options. The abdominal sacrocolpopexy achieves excellent correction of vaginal vault prolapse with minimal morbidity. PMID- 11383993 TI - Repair of enterocele and vault prolapse: transvaginal culdosuspension. AB - PURPOSE: Transvaginal culdosuspension for treatment of enterocele and vaginal vault prolapse is described. Surgical principles include high ligation of the hernia sac, obliteration of the cul-de-sac, and support of the vaginal cuff high on the levator plate. The normal vaginal axis is restored, and adequate vaginal depth is provided for normal sexual activity. MATERIALS AND METHODS: One hundred four patients underwent transvaginal culdosuspension in conjunction with enterocele repair (64 patients) or vaginal hysterectomy (40 patients). Two culdosuspension sutures support the vaginal vault to the origin of the sacrouterine and cardinal ligaments, and the cul-de-sac is obliterated with two pursestring sutures. RESULTS: One hundred patients were followed-up for a mean of 17.3 months. Recurrent vault prolapse or enterocele occurred in four patients. Complications were rare, and there were no instances of vaginal foreshortening, urinary retention, vaginal skin necrosis, bladder perforation, or rectovaginal fistula. CONCLUSIONS: Transvaginal culdosuspension is a safe and effective surgery for enterocele and vaginal vault prolapse. This technique restores the normal vaginal depth and axis, resulting in a sexually functional vagina. PMID- 11383994 TI - Colpocleisis for the treatment of vaginal vault prolapse. AB - Most females with total vault prolapse undergo reconstructive procedures to restore normal anatomy and function; however, elderly patients who no longer desire sexual intercourse or are medically unstable can be treated effectively with a vaginal closure or colpocleisis. The traditional approach to colpocleisis has been to simply invert the vagina using pursestring sutures after removing the vaginal mucosa. Although simple to perform, after repairing referred treatment failures who used this approach, we began to use a different approach that emphasizes the strength of an anterior repair and extensive posterior repair that then is sutured together. This vaginal closure is reinforced with a strong perineorrhaphy. This multicompartment colpocleisis was performed in 38 elderly females (mean age 77, range 68 to 88) with total vault prolapse. No treatment failures were noted with a mean follow-up of 24 months (range 3 to 52 months), and all patients were satisfied with the results of the procedure. No significant complications occurred, and no patient has regretted the loss of sexual function. The aim of this article is to discuss the indications, procedural aspects, and results of performing a multicompartment colpocleisis and partial colpocleisis for total vault prolapse in elderly females. PMID- 11383995 TI - Posterior vaginal wall prolapse: transvaginal repair of pelvic floor relaxation, rectocele, and perineal laxity. AB - PURPOSE: Posterior vaginal wall laxity is one manifestation of pelvic organ prolapse in the female. Recognition and repair of the inherent anatomical defects present in this condition are essential in order to ensure a satisfactory surgical result. METHODS AND MATERIALS: A successful operation for posterior vaginal wall prolapse will often involve repair of three discreet abnormalities in support of the posterior vaginal wall, including the pelvic floor, posterior vaginal wall fascia, and perineal musculature. An overaggressive repair is to be assiduously avoided as this can lead to excessive narrowing of the vaginal canal and considerable postoperative symptoms including dyspareunia. RESULTS: Durable restoration of anatomical support can be achieved in >80% of cases. Functional results in symptomatic patients undergoing posterior vaginal wall prolapse repair do not appear to be as successful in some areas. CONCLUSIONS: Successful surgical repair of posterior vaginal wall prolapse requires a thorough understanding of the anatomy and pathophysiology involved in this condition. A careful anatomical dissection and reconstruction will result in successful anatomical repair in the majority of patients with minimal morbidity. PMID- 11383996 TI - Evaluation and management of female urethral diverticulum. AB - The urethral diverticulum has many varied presentations; therefore, ultimate diagnosis may be difficult. Until recently, radiographic evaluation was difficult to perform, was uncomfortable for the patient, and had poor sensitivity. The increasingly widespread use of magnetic resonance imaging coupled with heightened awareness of the problem has enhanced the overall detection of urethral diverticula. Management is still primarily surgical and entails proper anatomical identification of the defects that cause the diverticulum, so subsequent reconstruction can be performed easily and with minimal morbidity. PMID- 11383997 TI - Male slings for postprostatectomy incontinence. AB - PURPOSE: Over the past few years, there has been increasing interest in using male slings for postprostatectomy incontinence (PPI). Currently, three different forms of the male sling has been described: one using synthetic materials and two using human fascia or dermis. This article will give a historical perspective on the male sling and describe the surgical techniques and early results using two different types of male slings using fascia or dermis. MATERIALS AND METHODS: From 1997 to 1999, nine patients with PPI underwent a bladder neck sling procedure using a combined perineal and abdominal approach. All patients had a suprapubic incision to expose the rectus muscle and to place a suprapubic tube. Since 1999, we have used a wide cadaveric fascial or dermal sling placed at the proximal bulbar urethra. Using an entirely perineal approach, the sling ends are fixated to the inferior pubic rami using bone anchors. RESULTS: Of 9 patients with bladder neck slings, 6 are dry, 1 is significantly improved, and 2 have failed, at a mean follow-up of 13 months. For the perineal male sling, 26 (45%) of 58 patients were completely dry, and overall 47 (81%) of 58 were significantly improved or were dry at a mean 6 months postoperatively. Neither group had significant complications. CONCLUSIONS: Both male slings appear to be safe and effective procedures for treating PPI; however, longer follow-up and additional experience are needed to determine their true role in the treatment of PPI. PMID- 11383998 TI - Current status of fascia lata allograft slings treating urinary incontinence: effective or ephemeral? AB - PURPOSE: The aim of this article is to review the current state and technique of pubovaginal sling construction using cadaveric allograft fascia lata. MATERIALS AND METHODS: A MEDLINE search was conducted to identify articles in the current literature addressing applications of cadaveric allograft fascia lata for surgical reconstructive procedures. RESULTS: Current series evaluating cadaveric fascia lata allografts for pubovaginal sling find equivalent short-term outcomes when compared to autograft fascia. Long-term follow-up is not yet available for these series. Variability in tissue processing or host factors may account for some failures with cadaveric fascial slings. CONCLUSIONS: Cadaveric fascia lata allografts are associated with reasonable efficacy and safety. Longer follow-up is needed. Further study of the relationship between tissue processing methods, surgical techniques, host factors, and outcome is indicated to better assess the long-term role of cadaveric fascia lata allograft for pubovaginal sling. PMID- 11383999 TI - Polypropylene sling for treatment of stress urinary incontinence: an alternative to tension-free vaginal tape. AB - PURPOSE: Sling procedures have been used successfully to treat stress urinary incontinence (SUI). We report our initial experience with the use of a synthetic polypropylene mesh for treatment of SUI. Based on similar surgical principles of cadaveric fascia sling, we describe placement of a thinly woven polypropylene mesh under the distal urethra. We describe our technique and report our initial results. MATERIALS AND METHODS: There were 146 consecutive patients evaluated. All patients had clinical evidence of SUI. Patients underwent preoperative evaluation with video urodynamics, symptom questionnaire, and cystoscopy. Postoperatively the patients were evaluated at 3-month intervals by symptom questionnaire, physical examination, and postvoid residuals (PVR). One year after the procedure, all patients were asked to fill out a voiding dysfunction and incontinence symptom questionnaire and a validated quality-of-life questionnaire. A flow and PVR also were obtained. RESULTS: Average intraoperative time was 27 minutes for the sling procedure. There were no intraoperative complications and one major postoperative complication. There was no permanent retention and no erosions. Ninety-two percent of patients had either no or rare stress incontinence. Postoperatively, 7% of patients developed de novo urge incontinence. CONCLUSION: We describe excellent results with a new simple, quick, and inexpensive method to correct SUI by placing a polypropylene mesh under the distal urethra. PMID- 11384000 TI - Tension-free vaginal tape and percutaneous vaginal tape sling procedures. AB - PURPOSE: Midurethral synthetic sling procedures for treatment of stress urinary incontinence (SUI) are gaining increased attention from surgeons specializing in female pelvic reconstructive techniques seeking successful patient outcomes through reproducible simplicity. This report describes the procedural steps and methods used to maximize the potential for successful outcomes using techniques of midurethral synthetic sling placement. Reported complications and surgical outcomes are reviewed with respect to patient selection and minimizing the potential for morbidity and mortality as long-term clinical experience is accumulated. MATERIALS AND METHODS: Tension-free vaginal or transvaginal tape (TVT) and the recently introduced percutaneous vaginal tape (PVT) are two new procedural choices for placement of synthetic sling material at the midurethra. Both procedures use sling material composed of polypropylene mesh, a nonabsorbable synthetic material, placed at the level of the midurethra via an antegrade (PVT, using a percutaneous ligature carrier) suprapubic approach or retrograde (TVT, using vaginal trocars) vaginal approach. Patient selection, procedural techniques, and methods described are based on observations obtained or reported from clinical experience. Outcomes and complications for TVT are derived from a literature review of all published articles in Index Medicus from 1996 to 2000. RESULTS: The experience with TVT for the last 5 years is encouraging. At 3-year follow-up for TVT, reported cure rates for SUI range from 80% to 95%. A multitude of worldwide reports on PVT with shorter follow-up support the findings of the TVT experience. Reproducible findings with midurethral synthetic slings are the short operative times recorded for the sling procedure, ease of technical performance, minimal patient discomfort, and a high rate of early return of normal voiding function. The rate of complications such as obstructive voiding or de novo instability (0-15%) and urinary retention necessitating a secondary procedure (2-4%) appear to be equal to the rates of contemporary competing technologies and procedures for treatment of SUI. CONCLUSIONS: Preliminary reports and the experience at our institution suggest that the techniques of midurethral synthetic sling placement of TVT and PVT are reproducible, easy to master, and minimally invasive with respect to tissue handling. Although complications with all anti-incontinence procedures exist, understanding the anatomical considerations and methodology of these unique procedures should minimize patient morbidity, avoid patient mortality, and produce a high rate of durable success. PMID- 11384001 TI - Aging and vision impairment research--facing the new challenges. PMID- 11384002 TI - The Smith-Kettlewell Institute (SKI) longitudinal study of vision function and its impact among the elderly: an overview. AB - We present an overview of a multifaceted longitudinal study of vision function and its interaction with daily activities, health, and well-being among 900 persons aged 58 to 102 years at the first visit. Standard vision measures as well as nonconventional tests designed to assess visual performance under the nonideal conditions encountered in everyday life were used. Here we summarize a few of the findings to date, with an emphasis on a direct comparison of declines in different aspects of vision function with age. The rates of declines with advancing age vary widely for the different vision functions. Also described is the reading performance of the sample and its association with some of the vision measures. Furthermore, we describe some of the associations between vision test scores and extensive longitudinal health and functioning data collected by the Buck Center for Research in Aging. Findings show that many older people with good acuity are effectively visually impaired in performing everyday tasks involving low and changing light levels, stereopsis, glare, and low contrast. We also found that vision under nonideal conditions cannot be predicted from standard acuity on an individual basis. PMID- 11384003 TI - Estimating sound source distance with and without vision. AB - BACKGROUND: Visual capture is an important perceptual phenomenon in which the spatial location of a visual target influences the perceived location of a related auditory target. Little is known about visual capture in a distance dimension. METHODS: Two groups of listeners judged the apparent distances of five loudspeaker sound sources, extending from 1 to 5 m. In one group, each listener was allowed to view the loudspeaker array. In the second group, listeners were blindfolded for the duration of the experiment. RESULTS: No visual capture effects were observed. Instead, the addition of vision was found to both improve distance judgment accuracy and lower judgment variability compared with the auditory-only stimulus. Auditory-only accuracy was found to substantially improve over the course of the experiment, however. CONCLUSIONS: Visual capture in distance is perhaps less general than suggested by past research, a result that has important implications for the display of spatial layout under conditions where vision is either missing or degraded. PMID- 11384004 TI - Feeling with the mind's eye: the role of visual imagery in tactile perception. AB - Cross-modal interactions are characteristic of normal perception. In this article, we discuss our work on cross-modal interactions between touch and vision in normally sighted humans. A region of extrastriate visual cortex, near the parieto-occipital fissure, is not only active during but also necessary for tactile discrimination of grating orientation (but not spatial frequency). This is consistent with a role for visual imagery in certain aspects of tactile perception. These findings have implications for the interpretation of visual cortical involvement in Braille reading by the blind. PMID- 11384005 TI - Navigating without vision: basic and applied research. AB - We describe some of the results of our program of basic and applied research on navigating without vision. One basic research topic that we have studied extensively is path integration, a form of navigation in which perceived self motion is integrated over time to obtain an estimate of current position and orientation. In experiments on pathway completion, one test of path integration ability, we have found that subjects who are passively guided over the outbound path without vision exhibit significant errors when attempting to return to the origin but are nevertheless sensitive to turns and segment lengths in the stimulus path. We have also found no major differences in path integration ability among blind and sighted populations. A model we have developed that attributes errors in path integration to errors in encoding the stimulus path is a good beginning toward understanding path integration performance. In other research on path integration, in which optic flow information was manipulated in addition to the proprioceptive and vestibular information of nonvisual locomotion, we have found that optic flow is a weak input to the path integration process. In other basic research, our studies of auditory distance perception in outdoor environments show systematic underestimation of sound source distance. Our applied research has been concerned with developing and evaluating a navigation system for the visually impaired that uses three recent technologies: the Global Positioning System, Geographic Information Systems, and virtual acoustics. Our work shows that there is considerable promise of these three technologies in allowing visually impaired individuals to navigate and learn about unfamiliar environments without the assistance of human guides. PMID- 11384006 TI - Low-vision patients with age-related maculopathy read RSVP faster when word duration varies according to word length. AB - Normally sighted younger and elder subjects as well as subjects with central visual field loss (CFL) from age-related maculopathy read rapid serial visual presentation (RSVP) text with words presented at a constant rate and at three different rates where word presentation duration varied according to word length. The elder subjects reading sentences foveally read fastest when word duration was constant. The younger group reading random words peripherally read faster at a variable word duration rate. The subjects with CFL read sentences an average of 33% faster when the presentation rate varied with word length. There was a trend for slow readers with CFL to benefit more than fast readers with CFL. We conclude that varying word duration based on word length in rapid serial visual presentation reading would improve reading rates for low-vision patients with CFL. PMID- 11384007 TI - The development of the Ocutech VES-autofocus telescope and a future binocular version. AB - Conventional optical low-vision devices are hampered by the physical constraints of magnification-shallow depth of field and narrow field of view, and for telescopes, the need to change focus for different distances. These characteristics make low-vision telescopic aids difficult to use, especially at important midrange distances. The Ocutech VES Autofocus bioptic telescope (VES AF) funded with National Institutes of Health-Small Business Innovation Research and Ontario (Canada) Ministry of Health grants, eliminates the need to manipulate the device to maintain focus. Clinical experience with the VES-AF has shown that autofocus (AF) enhances the acceptance and utilization of telescopic devices. Difficulty ignoring the fellow eye while sighting through the device has been a complaint of users that has undermined device acceptance. Although occlusion is an option to alleviate the diplopia experienced, the development of a binocular version may further enhance the acceptance, adaptation to, and utilization of telescopic devices and may offer wider fields of view as well. PMID- 11384008 TI - Vision multiplexing: an engineering approach to vision rehabilitation device development. AB - Multiplexing is the transmission of two or more messages simultaneously over the same communication channel in a way that enables them to be separated and used at the receiving end. The normal visual system provides us with a very wide field of view at an apparent high resolution. The wide field of view is continuously monitored at a low resolution, providing information for navigation and detection of objects of interest. These objects of interest are sampled over time using the high-resolution fovea. Most disabling visual conditions impact only one of the components, the peripheral low-resolution wide field or the central high resolution fovea. The loss of one of these components prevents the interplay of central and peripheral vision needed for normal function and causes disability. Traditionally low-vision aids replace or supplement the missing function, but usually at a cost of a significant loss in the surviving function. For example, magnifying devices increase resolution but reduce the field of view, whereas minifying devices increase the field of view but reduce resolution. A proposal to resolve many of the problems of current visual aids by exploring a general engineering approach--vision multiplexing--that takes advantage of the dynamic nature of human vision is presented. Vision multiplexing seeks to provide both the wide field of view and the high-resolution information in ways that could be accessed and interpreted by the visual system. This paper describes the use of optical methods and computer technologies in the development of a number of new visual aids, all of which apply vision multiplexing to restore the interplay of central and peripheral vision using eye movements in a natural way. PMID- 11384009 TI - Reading performance in older adults with good acuity. AB - PURPOSE: This study evaluated factors affecting reading performance in a sample (N = 544) of older adults (mean age 72.8 years, range 58 to 102) with good high contrast acuity (> or = 20/32). METHODS: Using the Pepper Reading Test, the relationship between reading rate and several vision measures was assessed. RESULTS: Mean corrected reading rate fell substantially over the age range tested despite the fact that these individuals all had good acuity. However, multiple regression analysis indicated that when other measures were taken into account (most notably, low-contrast vision, motor ability, and attentional field integrity), age was not a significant independent predictor of corrected reading rate. CONCLUSION: Reading is an important skill, and ways of enhancing reading performance should be explored. Good high-contrast acuity does not assure that older individuals can read satisfactorily. PMID- 11384010 TI - Eye movements and reading with large print and optical magnifiers in macular disease. AB - PURPOSE: Reading rate has been the main performance measure in studies that have compared reading with large print and optical magnifiers; eye movement characteristics have not been considered. We compared both eye movement characteristics and reading rates for subjects with macular disease reading without and with a range of low-vision devices. METHODS: Silent reading rate and eye movement characteristics for text passages at critical print size of 21 subjects aged 14 to 88 years with macular disease were measured with and without their preferred low-vision device. Saccadic frequency was determined from a sequencing task comprising five letters each separated by 5 degrees. Eye movements were recorded using an infrared limbal reflection system. RESULTS: There were no significant differences in reading rate, fixation durations, saccade numbers per word, or percent retrace time when using a low-vision device compared with reading without a low-vision device. The percentage of regressions was, however, lower with the low-vision device. Saccadic frequency in the sequencing task was predictive of reading performance with and without a low vision device. CONCLUSIONS: When reading at critical print size, in terms of reading rate or saccades per word, there was no advantage to using large print over an optical low-vision device. PMID- 11384011 TI - Quality of life of low-vision patients and outcomes of low-vision rehabilitation. AB - PURPOSE: The impact of low vision on self-reported quality of life and changes after low-vision intervention are investigated. METHODS: Literature reports from 1990 to 2000 are reviewed. RESULTS: Low vision is associated with increased risk for depression and decreased functional status and quality of life. Decreased visual acuity, visual field loss, and occasional blurred vision are also associated with decreased quality of life. Improvements in both functional status and quality of life occur after low-vision service delivery. CONCLUSIONS: Self reported quality of life is a significant outcome measure for low-vision rehabilitation. Questionnaires that are more sensitive to rehabilitation services provided as well as patient needs and goals are required to facilitate development of rehabilitation plans and to compare techniques, devices, and programs. Attention should be given to measurement properties, validity, and reliability of instruments used currently and in development of new instruments. PMID- 11384012 TI - Comparison of driving performance of young and old drivers (with and without visual impairment) measured during in-traffic conditions. AB - PURPOSE: The aim of this study was to determine whether visual impairment and age affected driver performance and safety rated during in-traffic driving conditions. METHODS: Participants included 30 younger, 25 middle-aged, and 35 older subjects with normal vision and 47 older subjects with visual impairment. All subjects were legally eligible to drive. Driving performance was assessed during in-traffic conditions by a professional driving instructor and an occupational therapist using specific scoring criteria. RESULTS: Group allocation significantly affected driving performance as assessed by the driving instructor and occupational therapist (p < 0.05). The driving instructor and occupational therapist scores were highly correlated (r = 0.76). Of the drivers who were scored as being unsafe, all were older and the majority (75%) had visual impairment. DISCUSSION: Older drivers with or without visual impairment were rated as being less safe than the younger and middle-aged drivers with normal vision. PMID- 11384013 TI - Timed instrumental activities of daily living tasks: relationship to visual function in older adults. AB - PURPOSE: To identify instrumental activities of daily living (IADL) tasks whose completion time is related to visual function in older adults. METHODS: Visual function (acuity, contrast sensitivity, and useful field of view) and the time to complete 17 visual tasks of everyday life were measured in a sample of 342 older adults (mean age 71 years, range 56 to 86) recruited from eye clinics. The timed IADL (TIADL) tasks included a variety of visual activities such as reading ingredients on cans of food and instructions on medicine bottles, finding a phone number in a directory, locating items on a crowded shelf and in a drawer, and using a screwdriver. RESULTS: Multiple regression analysis indicated that poorer scores for acuity, contrast sensitivity, and useful field of view were independently associated with longer times to complete visual TIADL tasks, even after adjusting for age, educational level, depression, and general health. Cognitive status also had a significant, independent association with timed task performance. CONCLUSIONS: Older adults' timed performance in everyday tasks is related to various aspects of visual function independent of the influences of other functional and health problems and advanced age. This suggests that TIADL tasks may eventually be useful as performance outcomes in intervention evaluations targeted at reversing vision impairment or minimizing its impact. To understand the relationship between vision impairment and TIADL task performance in older adults, cognitive impairment needs to be taken into account because it has a relatively strong and independent relationship with visual TIADL task performance. PMID- 11384014 TI - Presidential address. PMID- 11384015 TI - Valedictory address. PMID- 11384016 TI - Staff shortages. PMID- 11384018 TI - Patient care. PMID- 11384017 TI - Dental implants. PMID- 11384019 TI - A culture for learning. PMID- 11384020 TI - Ridge augmentation using mandibular tori. AB - A 19-year-old female was referred by her dental practitioner for the restoration of missing maxillary lateral incisors and canines. Ridge augmentation was required. This was undertaken using mandibular tori as the sites for harvesting bone. The grafting was successful and the spaces were subsequently restored using resin-bonded bridgework. The case reports that mandibular tori provide a local and convenient source of bone for ridge augmentation procedures. PMID- 11384021 TI - Developing an index of restorative dental treatment need. AB - The process undertaken to establish an initial pilot index for restorative dental treatment is described. Following consultation with a wide range of clinicians and others, an outline framework for the index was developed and comprised three main components: 1. Patient identified need for treatment: the data from the patient perceived need questionnaire were inconclusive; 2. Complexity of treatment (assessed by clinicians): this was found to be a practical tool capable of being used by a range of dentists. A booklet has been produced which describes the process of using the scoring system; 3. Priority for treatment (assessed by clinicians): three levels of priority were identified; the highest priority was assigned to patients with inherited or developmental defects that justify complex care (eg clefts of the lip and palate). The initial development of the index has had some success in a difficult area. The treatment complexity component is the most developed and may allow both referrers and commissioners of specialist restorative dentistry to determine appropriate use of skilled clinicians' expertise. PMID- 11384022 TI - Screening panoramic radiology of adults in general dental practice: radiological findings. AB - AIM: To identify the radiological findings from routine screening panoramic radiographs taken of adult (> or = 18 years) patients in general dental practice. METHOD: Forty-one general dental practitioners (GDPs) who routinely took panoramic radiographs of all new adult patients were recruited. In total, they submitted 1,818 panoramic radiographs of consecutive patients along with basic patient information, radiological reports and treatment plans. The radiographs were also reported by 'experts' (consensus of two dental radiologists). Radiological findings were recorded from the GDP assessments (dentist RY), the experts (expert RY), after exclusion of findings that would have been seen on posterior bitewing radiographs (MRY) and after exclusion of findings of no relevance to treatment (MRYT). RESULTS: There was no significant difference in age profile between the study sample and Dental Practice Board population figures (P = 0.26). No radiographs other than the panoramic radiograph had been taken for 57.1% of patients. For the GDP assessments, only 4.6% of patients had radiographs with no radiological findings, while for the experts this proportion was 3.1%. With the exception of the assessment of periodontal bone loss, the experts diagnosed significantly greater proportions of cases as having positive radiological findings. Agreement between dentist and expert assessments varied greatly. When findings from bitewing radiographs were excluded, no radiological findings were recorded on the radiographs of 17.2% of patients. When proposed treatment plans were taken into account, the majority of patients' radiographs (56.3%) had no radiological findings of relevance to treatment. CONCLUSIONS: The choice of radiographic examination for the majority of patients in the study did not follow current guidelines. Dentists diagnosed fewer abnormalities than did experts. While many radiological findings are revealed by panoramic radiography, these may either duplicate information from bitewing radiographs or are often of no significance to treatment planning. This study did not provide evidence to support the practice of routine panoramic radiography of all new adult patients. PMID- 11384023 TI - The influence of two different dental local anaesthetic solutions on the haemodynamic responses of children undergoing restorative dentistry: a randomised, single-blind, split-mouth study. AB - OBJECTIVES: This investigation was designed to study the haemodynamic effects of two different local anaesthetic solutions during restorative dental treatment in children. DESIGN: A randomised, single-blind, split-mouth cross-over design was employed using children undergoing bilaterally similar restorative treatments over two visits. SETTING: The study was performed in a dental hospital paediatric dentistry department. METHODS: Ten children participated. At one visit the local anaesthetic was 2% lidocaine (lignocaine) with 1:80,000 epinephrine (adrenaline); at the other the anaesthetic was 3% prilocaine with 0.03IU/ml felypressin. Local anaesthetic was administered at a dose of 0.5 ml/10 kg body weight. Blood pressure and heart rate were measured before and during treatment with an automatic blood pressure recorder. Data were analysed by ANOVA and Student's paired t test. RESULTS: Significant differences between treatments in diastolic blood pressure (F = 2.37; P = 0.05) and heart rate (F = 2.98; P< 0.02) were noted. The heart rate increased ten minutes following the injection of the epinephrine-containing solution. The diastolic blood pressure fell 20 minutes after injection of lidocaine with epinephrine. CONCLUSION: The choice of local anaesthetic solution influences the haemodynamic response during restorative treatment in children. PMID- 11384024 TI - The use of real time video magnification for the pre-clinical teaching of crown preparations. AB - OBJECTIVE: To investigate the effect on the undergraduate learning process of using an alternative method designed to enhance the visual demonstration of taper on full veneer crown preparations (better understanding of the value of taper on preparations early in the teaching programme in restorative dentistry). DESIGN: A comparison between the conventional teaching of full veneer crown preparations and the same teaching with the additional use of a magnified real time video display using a surgical microscope was investigated in this study. OUTCOME MEASURES: The degree of taper was measured for replica full crown preparations and results compared between different cohorts of undergraduates and experimental conditions. RESULT: Undergraduates taught using the real time video produced more accurately tapered preparations. This ability was retained over one year. CONCLUSIONS: A possible explanation for the result was that the use of magnification improved the undergraduates' precise understanding of taper by enhancing their ability to evaluate this critical measurement during the teaching process. PMID- 11384025 TI - Lactobacillus crispatus and its nonaggregating mutant in human colonization trials. AB - A wild-type Lactobacillus crispatus, showing a cell aggregation phenotype and its spontaneous nonaggregating mutant were compared for their in vitro adhesion properties to human ileal mucus and to a cultured human colonic cell line (Caco2) and for their in vivo colonization and adhesion potential with colonoscopy patients as volunteers in feeding trials. The wild-type strain adhered better to mucus or to Caco2 cells than did the mutant. Altogether, three human trials with the wild type and two with the mutant strain were performed. In two of the trials, the wild type could be recovered from either fecal samples or biopsies taken from the colon, while the mutant strain could not be demonstrated in either of the trials where it was used. The L. crispatus colonies recovered from the trials were often mixed, and several enterococci and lactobacillus strains coaggregating with L. crispatus wild type could be isolated. The results indicate that the surface-mediated properties, such as aggregation, of lactobacilli can have a role in adhesion and colonization. PMID- 11384026 TI - Tyrosine and phenylalanine catabolism by Lactobacillus cheese flavor adjuncts. AB - Bacterial metabolism of Tyr and Phe has been associated with the formation of aromatic compounds that impart barny-utensil and floral off-flavors in cheese. In an effort to identify possible mechanisms for the origin of these compounds in Cheddar cheese, we investigated Tyr and Phe catabolism by Lactobacillus casei and Lactobacillus helveticus cheese flavor adjuncts under simulated Cheddar cheese ripening (pH 5.2, 4% NaCl, 15 degrees C, no sugar) conditions. Enzyme assays of cell-free extracts indicated that L. casei strains catabolize Tyr and Phe by successive, constitutively expressed transamination and dehydrogenation reactions. Similar results were obtained with L. helveticus strains, except that the dehydrogenase enzymes were induced during incubation under cheese-ripening conditions. Micellar electrokinetic capillary chromatography of supernatants from L. casei and L. helveticus strains incubated under simulated cheese-ripening conditions confirmed that Tyr and Phe transamination and dehydrogenation pathways were active in both species and also showed these reactions were reversible. Major products of Tyr catabolism were phydroxy phenyl lactic acid and p-hydroxy phenyl acetic acid, while Phe degradation gave rise to phenyl lactic acid, phenyl acetic acid, and benzoic acid. However, some of these products were likely formed by nonenzymatic processes, since spontaneous chemical degradation of the Tyr intermediate p-hydroxy phenyl pyruvic acid produced p-hydroxy phenyl acetic acid, p-hydroxy phenyl propionic acid, and p-hydroxy benzaldehyde, while chemical degradation of the Phe intermediate phenyl pyruvic acid gave rise to phenyl acetic acid, benzoic acid, phenethanol, phenyl propionic acid, and benzaldehyde. PMID- 11384027 TI - Recombinant lamb chymosin as an alternative coagulating enzyme in cheese production. AB - Recombinant lamb chymosin (RLC) was prepared and tested for its potential use in cheese production. The milk clotting activity and proteolytic activity of RLC were evaluated in comparison with commercial recombinant calf chymosin (RCC), cow rennet (CR), and microbial coagulant (MC). RLC, RCC, and MC showed similar responses to pH, with a sharp increase of the coagulation time at pH 6.6 to 6.8 and decrease of curd firmness at the pH 6.5 to 6.6. In the case of CR, we observed two clear increases in the coagulation time and decreases in the curd firmness, at pH 6.4 to 6.5 and 6.6 to 6.8. Optimal clotting activity was obtained for RLC at 40 degrees C, for both CR and RCC at 45 degrees C, and for MC at 60 degrees C. The temperature instability of RLC at temperatures above 45 degrees C could constitute a benefit in making hard cheese varieties. The additon of CaCl2 to milk resulted in enhanced clotting activity of all coagulants, most prominently for CR. The proteolytic activity of RLC was significantly lower from that of CR but not significantly different from the activity of RCC. The lower proteolytic activity in the cheese made with RLC did not have negative effect on organoleptic properties. The overall quality of the cheese made with RLC was at least comparable to that of the cheese made with RCC, and both cheeses were better scored than the cheese made with CR. PMID- 11384028 TI - Effect of rennet coagulation time on composition, yield, and quality of reduced fat cheddar cheese. AB - This study compared the effect of coagulum firmness at cutting on composition of 50% reduced-fat Cheddar cheese. Coagulum firmness was determined by subjective evaluation by the cheese maker. Three firmness levels were tested, and these corresponded to average times of coagulant addition to cutting the curd of 25, 48, and 65 min. A slow acid-producing culture was used, and ripening times were altered to give similar curd pH values throughout cheese making. A longer rennet coagulation time (firmer coagulum at cutting) resulted in an increase in cheese moisture as well as an increase in cheese yield. The percentages of fat recovered in the cheese decreased with increasing curd firmness. The percentage of nitrogen recovered in the cheese was similar among the treatments. The amount of whey collected from the curd after milling increased as the coagulum firmness at cutting increased. Higher moisture content and lower pH of cheese made from the firmer curd at cutting contributed to softer, smoother-bodied cheeses, but the Cheddar flavor intensity was not affected. PMID- 11384029 TI - Lipolysis of butter oil by immobilized lamb pregastric esterase: I. Uniresponse kinetics-pH and temperature effects. AB - Lamb pregastric esterase, immobilized by physical adsorption on microporous polypropylene in a hollow fiber reactor, has been employed to effect the continuous hydrolysis of the triglycerides in butter oil. Experimental data were obtained at temperatures from 35 to 45 degrees C and pH values from 5.5 to 6.5. The overall rate of hydrolysis was fastest at 40 degrees C and a pH of 6.0. Nonlinear regression methods were employed to determine the kinetic parameters of rate expressions based on a generic Ping-Pong Bi Bi mechanism. The best nonlinear fit of the data was consistent with a mechanism that assumes that acylation of the enzyme is the rate-limiting step in the hydrolysis reaction. PMID- 11384030 TI - Effect of region and herd size on dairy herd performance parameters. AB - Differences in selected Dairy Herd Improvement (DHI) performance parameters among regions and herds of different size categories were evaluated. DHI records from herds in 37 states were grouped into North, Midsouth, and South regions, and six herd size categories (20 to 49, 50 to 99, 100 to 149, 150 to 249, 250 to 449, and > or = 450 cows). The North region had higher income over feed costs (IOFC); milk, fat, and protein rolling herd averages; summit milk; standardized 150-d milk; and percentage of cows in milk, than the other regions. These variables were lowest for the South region. Cost/45.4 kg of milk, days open, days dry, and somatic cell counts were lowest in the North region and were highest in the South. Percentage of cows entering and leaving the herd were highest in the Midsouth and were lowest in the South. Larger herds had higher total feed cost, IOFC, milk, fat, and protein rolling herd averages, summit milk, standardized 150 d milk, percentage of cows entering and percentage leaving the herd than smaller herds. Larger herds had lower somatic cell counts than smaller herds. Cost/45.4 kg of milk, days dry, days open, days in milk, and percentage of cows in milk did not show clear trends among different herd sizes. There were significant interactions between region and herd size for some of the variables. PMID- 11384031 TI - Stage of cycle, incidence, and timing of ovulation, and pregnancy rates in dairy cattle after three timed breeding protocols. AB - In experiment 1, 705 cows were assigned to three treatments: 1) the Ovsynch protocol (a GnRH injection given 7 d before and another 48 h after one PGF2alpha injection); 2) PGF2alpha + Ovsynch (one PGF2alpha injection given 12 d (d -22) before initiating Ovsynch (d -10); and 3) 2xPG12 (two PGF2alpha injections 12 d apart; d -15 and -3, followed 48 h later by a GnRH injection. All cows were inseminated (d 0) 16 to 20 h after the GnRH injection on d -1. Cyclic status was estimated by serum progesterone. More cows were in early diestrus at d -10 for PGF2alpha + Ovsynch (36%) and 2 x PG12 (29%) versus Ovsynch (19%). Multiparous cows receiving PGF2alpha + Ovsynch had greater pregnancy rates via ultrasonography at d 28 after AI (42%) than contemporaries after Ovsynch (28%) or 2xPG12 (27%) but did not differ significantly at palpation 10 to 30 d later (28, 19, and 17%, respectively). Pregnancy of first-parity cows was similar across treatments at 28 d (41%) or at palpation (33%). Pregnancy rates for 128 anestrous cows were lower, regardless of treatment. Overall embryo survival from d 28 until palpation was 72% but was only 44% in 2xPG12 cows that were anestrus through d 10. Experiment 2 included the three treatments above plus controls (one GnRH injection 7 d before PGF2alpha and AI after estrus). Preovulatory follicles were 6 to 11% larger near estrus in controls than on d -1 in cows receiving GnRH. More controls ovulated by 32 h after onset of estrus than were treated cows by 32 h after GnRH, but percentages (79 to 94%) were similar by 40 h. In multiparous cows, PGF2alpha before Ovsynch increased pregnancy rates, whereas the 2xPG12 protocol produced similar pregnancy rates as Ovsynch across parities. Ovulation was effectively induced by 40 h after GnRH. PMID- 11384032 TI - Influence of concentrate allocation and energy balance on postpartum ovarian activity in Norwegian cattle. AB - The effects of different levels of concentrate, milk yield, and energy balance (EB) on postpartum ovarian activity were studied in 164 lactations from 108 moderate yielding, dual-purpose cows. All cows were of the breed Norwegian Cattle. The cows were assigned to three different concentrate allocations and had free access to roughage. Cows in the high energy group were fed as suggested by Norwegian standards, whereas cows in the low energy group were supplied with approximately 40% less concentrate than recommended. Cows in the medium energy group were fed as the low energy group prepartum and as the high energy group during the first 90 d in milk (DIM). Energy balance was estimated by subtracting energy required for maintenance and lactation from energy intake. Ovarian activity was expressed in categories (early, mid and late responders) of days to first luteal response (milk progesterone concentration >5 ng/ml) and days to first regular cycle. Energy balance and milk yield throughout the first 100 DIM were compared between categories of concentrate allocation and ovarian activity using repeated measures mixed models. Decreasing level of concentrate was related to negative EB and declining milk yield. Both negative EB and high milk yield were associated with delayed postpartum ovarian activity. Concentrate allocation did not, however, alter days to first luteal response or days to first cycle. This study demonstrated that although negative EB was associated with delayed ovulation, Norwegian Cattle adapted to less energy-dense rations by reducing milk yield rather than delaying onset of ovarian activity. PMID- 11384033 TI - Effects of thyroid hormones on bovine granulosa and thecal cell function in vitro: dependence on insulin and gonadotropins. AB - The objective of this study was to evaluate the influence of thyroxine (T4) and triiodothryonine (T3) on steroid production by bovine granulosa and thecal cells. Granulosa and thecal cells were obtained from small (1 to 5 mm) and large (> or = 8 mm) follicles of cattle, respectively, and cultured for 4 d. We conducted six experiments to evaluate the effect of 2 d of exposure to various doses of T3 or T4. In insulin- or insulin plus FSH-treated granulosa cells of experiment 1, 30 and 100 ng/ml of T4 had no effect on aromatase activity or progesterone production. In experiment 2, in the presence of insulin and FSH, 1 and 3 ng/ml of T3 weakly (<1.4-fold) increased aromatase activity of granulosa cells but had no effect on progesterone production. Low doses of T4 (3 to 30 ng/ml) tested in experiment 3 had no effect on aromatase activity but increased (to as much as 1.4 fold) progesterone production by granulosa cells. In experiment 4, T4 (30 ng/ml) increased (to 1.2-fold) progesterone production by granulosa cells only in the presence of FSH and had no effect on aromatase activity. In thecal cells of experiment 5, in the presence of insulin and LH, 30 and 100 mg/ml of T4 increased androstenedione production to 2.3- and 2.8-fold, respectively; only 100 ng/ml of T4 was effective at stimulating progesterone production by thecal cells. In experiment 6, 1 ng/ml of T3 increased thecal cell androstenedione production to 3.9-fold, whereas 3 ng/ml of T3 was without effect; progesterone production was not affected by T3. These results support the hypothesis that thyroid hormones may have direct stimulatory effects on ovarian function in cattle, acting at the level of granulosa and thecal cells. PMID- 11384034 TI - Cell subpopulations and cytokine expression in cow milk in response to chronic Staphylococcus aureus infection. AB - Staphylococcus aureus is a major pathogen in bovine intramammary infections of subclinical and chronic nature. Persistent infection with S. aureus has been postulated to be associated with an impaired immune response. This study was designed to define changes in peripheral blood and milk cell subpopulations during chronic S. aureus infection. The expression of specific antigens on the surface of lymphocytes and neutrophils was studied by flow cytometry. Cytokines and cytokine transcripts elaborated by the milk-derived cells were also investigated, using ELISA and reverse transcription polymerase chain reaction, respectively. The results indicated that cell subpopulations in blood from infected cows were not modified. In contrast, changes occurred in infected milk: neutrophils were the main cell population, but they were not in a highly activated state; the CD8+ T-lymphocytes were mainly recruited compared with the CD4+ T-lymphocytes, suggesting that CD8+ T-lymphocytes play an important role in chronic S. aureus infection. Also, the proportion of the B-lymphocytes among the total lymphocyte population was increased, suggesting that a humoral response developed, and no change was observed in the gammadelta subset. No cytokine mRNA was found in milk cells from uninfected mammary glands. In contrast, interleukin (IL)-1alpha, IL-1beta, IL-6 and tumor necrosis factor alpha pro-inflammatory cytokine and IL-10 and IL-12 regulatory cytokine mRNA were synthesized in cells derived from infected mammary glands, whereas no IL-2 nor IL-4 mRNA were found. Therefore, cells present in milk during chronic S. aureus infection were activated, but did not reveal any polarization of the immune response. PMID- 11384035 TI - Effect of monensin on blood ketone bodies, incidence and recurrence of disease and fertility in dairy cows. AB - Twelve trials designed to test the effect of monensin on milk production were carried out at eight different research farms. Data from these studies were evaluated for effects of monensin on health and reproduction. Monensin was added to the concentrate starting either 2 wk before (293 cows) or 5 wk after calving (601 cows) for periods ranging 16 to 37 wk. Applied after calving, the incidence of clinical mastitis was reduced in the monensin-treated animals. Monensin decreased the rate of intramammary infection (approximated by a change from below to above 250,000 somatic cells in milk) in first lactation heifers by 13%. Indicated by an adjusted incidence rate ratio of 0.58 (P = 0.03), the case incidence of noninfectious lameness decreased from 31% in control cows to 18% in cows receiving monensin before calving. The time from calving to first service was shorter for cows fed monensin before calving. Monensin reduced calving to conception intervals in cows with endometritis during the pretreatment period. No significant effects of monensin were observed for the duration of intra mammory infection (somatic cells in milk above 250,000), infectious lameness, endometritis, cystic ovarian disease, a summary category of 'other diseases,' times from calving to first observed estrus, and from first service to conception. PMID- 11384036 TI - Effects of sustained release bovine somatotropin (sometribove) on animal health in commercial dairy herds. AB - The health of dairy cows given bovine somatotropin (bST) for one lactation was evaluated in 28 commercial herds located in four regions of the United States. At least six herds were in a region and at least one herd/region contained fewer than 60 cows. Cows (n = 1213) were assigned randomly to control or bST groups and were treated beginning in wk 9 to 10 of lactation and every 14 d until dry-off or d 400 of lactation. Management was according to site practices. Cows were observed for health-related signs by farm personnel daily and by the herd veterinarian biweekly. Average 305-d test-day milk yields were 932 kg greater for bST-treated cows. Pregnancy rates, days open, twinning, cystic ovaries, or abortions were unaffected by treatments. Supplementation of cows with bST had no effect on total mastitis cases, total days of mastitis, duration of mastitis, or the odds ratio of a cow to develop mastitis. Cows supplemented with bST used more medications for health events other than mastitis. This usage was associated primarily with treatments for disorders of the foot and hock. Supplemented cows had a slight increase in foot disorders. There was no effect of supplementation with bST on culling from the herd or removal from study. Overall, the results confirm that label directions for bST are adequate for safe use under field conditions. All clinical signs observed in this study occur normally in dairy herds and were managed in cows supplemented with bST. PMID- 11384037 TI - Development of a simple in vitro assay for estimating net rumen acid load from diet ingredients. AB - The objective of these experiments was to develop a simple in vitro technique for evaluating the production and neutralization of acid as feeds ferment in the rumen. An in vitro approach was adopted to eliminate animal factors. The procedure was based on the method of Tilley and Terry, with some modifications developed in this project. Residual acidity (acidogenicity value) was determined by the dissolution of Ca from CaCO3 powder added to the media at the end of 24-h incubations. Acidogenicity values (AV) were higher when 20% strength buffer was used, while lowering buffer pH increased values, equally across all feeds. There was no effect of donor animal diet, but considerable day-to-day variation in the fermentation activity of rumen fluid. This variation likely reflected the substrate preferences of differing microbial populations, so that several standard feeds may be required to account for this effect. A series of 28 diverse feed ingredients was evaluated for AV using a mixture design, with 85 combinations of ingredients: 100% of each ingredient (n = 28); 50% of each ingredient and an equal mixture of all others (n = 28); equal mixture of all ingredients, excluding one (n = 28); and an equal mixture of all ingredients (n = 1). The effects of most ingredients on AV were essentially linear, though some extreme ingredients showed nonlinear effects. Protein sources had low AV, forages intermediate AV and starchy feeds high AV. Calcium contained within feeds contributed to AV, particularly for legumes, sugar beet pulp, and citrus pulp, and must be accounted for. PMID- 11384038 TI - Cell release from alginate immobilized Lactococcus lactis ssp. lactis in chitosan and alginate coated beads. AB - The effects of chitosan and alginate coatings of alginate beads with entrapped Lactococcus lactis ssp. lactis were studied in batch and continuous fermentations. Chitosan coating reduced the final concentrations of free cells, the initial release of free cells and the rate of lactate production in milk fermented batch-wise to a final pH of 4.7 in five consecutive batch fermentations. An alternative experimental system based on continuous fermentation with controlled pH and a high dilution rate was developed to better study the phenomenon of cell release. To estimate the effects of different bead coatings on cell release, alginate beads were coated with chitosan or alginate, or sequentially with chitosan/alginate or chitosan/alginate/chitosan. Chitosan coating alone seemed to reduce the rate of cell release only in the early stages of the fermentation, while sequential coatings with chitosan and alginate showed significant reduction throughout the whole test period. To examine whether the observed effects of bead coating could be explained only by a decrease in cell activity, the ratios between the rate of cell release and the rate of lactate production were examined during the fermentations for the different beads. This ratio showed qualitatively the same behavior as direct results of volumetric cell release. PMID- 11384039 TI - Relationships between milk urea concentrations and nutritional management, production, and economic variables in Ontario dairy herds. AB - The objectives of this study were to describe the relationships between milk urea concentrations and nutritional management, production, and economic variables in commercial dairy herds. Dairy Herd Improvement (DHI) test-day milk urea data, production data, and information on ration nutrient composition and feeding management programs were collected over a 13-mo period from 53 commercial Ontario dairy herds. Economic variables included gross milk revenue, feed costs, and income over feed costs. Herd mean milk urea concentrations had a positive relationship with dietary levels of crude protein (CP), rumen degradable protein (RDP), and rumen undegradable protein (RUP) and a negative relationship with dietary levels of nonfiber carbohydrates (NFC), forage:concentrate (F:C) ratio, NFC:CP ratio, and NFC:RDP ratio. These findings are consistent with experimental studies that used chemical methods of milk urea analysis. Herd mean milk urea concentration was not associated with feeding management (e.g., total mixed rations, component feeding, feeding frequency, or synchrony of forage and concentrate feeding). Herd mean milk urea was not associated with either mean milk yield or linear score. Herd mean milk urea had a positive relationship with feed costs per cow per day but was not associated with gross milk revenue per cow per day. Herds with a high mean milk urea concentration tended to have lower income over feed costs per cow per day. High herd mean milk urea concentrations were associated with higher feed costs per kilogram of milk fat but lower gross milk revenue and lower income over feed costs per kilogram of milk fat. The results of this study demonstrate that DHI milk urea measurements produced by an infrared test method offer a useful tool for monitoring the efficiency of nitrogen utilization in commercial dairy herds. The results also suggest that diets may be balanced to achieve greater efficiency of nitrogen utilization, lower milk urea concentrations, and lower feed costs, while still achieving high milk production. This may lead to improved income over feed costs. PMID- 11384041 TI - The effect of Lactobacillus buchneri and other additives on the fermentation and aerobic stability of barley silage. AB - Whole-plant barley (39.4% dry matter) was treated with various chemical and biological additives to assess their effects on silage fermentation and aerobic stability. Treatments were untreated forage, forage treated with several amounts of Lactobacillus buchneri and enzymes (L. buchneri at 1 x 10(5), 5 x 10(5), and 1 x 10(6) cfu/g of fresh forage), forage treated with an inoculant containing (Lactobacillus plantarum, Pediococcus pentosaceus, Propionibacterium freudenreichii, and enzymes), or forage treated with a buffered propionic acid based additive (0.2% of fresh weight). Sixty-nine d after ensiling, silages treated with L. buchneri and enzymes had lower pH, but had higher concentrations of acetic and propionic acids and higher concentrations of ethanol when compared with untreated silage. Silage treated with the multistrain inoculant containing L. plantarum had lower pH and higher concentrations of lactic acid, but lower concentrations of ammonia-N, neutral detergent fiber, and acid detergent fiber than did untreated silage. The addition of the buffered propionic acid additive resulted in silage with higher concentrations of lactic and acetic acid compared with untreated silage. Numbers of yeasts in all silages were low at silo opening (less than 3.0 log cfu/g) and were numerically the lowest in silages treated with L. buchneri but only treatment with the intermediate and high level of L. buchneri improved the aerobic stability of silage. Because of the altered fermentation pattern, inoculation with L. buchneri, when applied at equal to or more than 5 x 10(5) cfu/g, and enzymes improved the aerobic stability of barley silage. PMID- 11384040 TI - Multiplex polymerase chain reaction assay for simultaneous detection of Staphylococcus aureus and streptococcal causes of bovine mastitis. AB - To improve diagnosis of mastitis in dairy cattle, a multiplex polymerase chain reaction (PCR) assay was developed for the simultaneous detection of the four major bacterial causes of bovine mastitis, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, and Streptococcus uberis. The target sequence was the 16S to 23S rRNA spacer regions. The performance of the assay was examined with 117 milk samples collected from a subclinically infected herd, and the diagnostic specificities and sensitivities of the multiplex PCR were compared with conventional culture. PCR was significantly more sensitive than culture for detection of S. aureus and S. uberis, but there were no significant differences in sensitivities between PCR and culture for the detection of S. agalactiae and S. dysgalactiae. The results suggest that this multiplex PCR assay could be used as an alternative method in routine diagnosis for rapid, sensitive, and specific simultaneous detection of S. aureus, S. agalactiae, S. dysgalactiae, and S. uberis in milk samples. PMID- 11384042 TI - Effects of forage source and amount of concentrate on rumen and intestinal digestion of nutrients in late-lactation cows. AB - The objectives of this study were to determine the effects of dietary forage source with two concentrate concentrations on dry matter (DM) intake, rumen fill, ruminal and intestinal digestibility of nutrients, and duodenal N fractions in lactating cows. Four rumen and duodenal cannulated Holstein cows in late lactation were used in 4 x 4 Latin square design experiment with 21-d periods. Diets were 1) 65% first-cut alfalfa silage and 35% concentrate, 2) 50% alfalfa and 50% concentrate, 3) 65% bromegrass silage and 35% concentrate, and 4) 50% bromegrass and 50% concentrate. Dry matter intake was not affected by forage source but tended to be (P = 0.08) higher for cows fed diets with 50% concentrate. Rumen fill was greater (P < 0.01) for cows fed bromegrass compared with those fed alfalfa silage. Ruminal and intestinal digestion of DM was not affected by dietary forage source or concentrate level. Total N intake was greater for cows fed alfalfa-based diets, reflecting the higher crude protein content of alfalfa. However, total N flow at the duodenum was not affected by either forage source or concentrate in the diet. Although forage source influenced the site of digestion of some nutrients no significant effects on total tract digestibilities were observed. PMID- 11384043 TI - Milk production of fall-calving dairy cows during summer grazing of grass or grass-clover pasture. AB - Milk production of fall-calving dairy cows during subsequent summer grazing was evaluated in two consecutive years using a total of 80 mid- to late-lactation Holsteins. Cows calved during September and October and grazed from April to August in the following year. In yr 1, 27 cows grazed a native grass pasture and 13 cows grazed a native grass-clover mixed pasture containing 26% red clover and white clover. In yr 2, 40 cows grazed native grass pasture as one group. Also, cows in yr 2 were administered bovine somatotropin, whereas in yr 1, no bST was used. Grazing cows also were fed concentrate supplements at 6.2 kg/d of dry matter (DM) in yr 1 and 7.9 kg/d of DM in yr 2 to provide 35 to 40% of total intake. Average daily milk during the grazing period decreased 3.6 kg in yr 1 and 7.7 kg in yr 2 when compared with milk yield extrapolated from the lactation curve established 10 wk before being turned out to pasture. Estimated DM intake during grazing was also less than what would have been expected had cows continued on a total mixed ration in confinement. Cows grazing the mixed pasture of grass and clover yielded 1.3 kg/d more milk than those grazing the grass pasture in yr 1. A decrease in milk resulting from the change from total mixed ration fed in confinement to grazing supplemented with concentrates was not avoided with these mid- to late-lactation cows, but the cumulative loss over the lactation was less than with early lactation cows in a companion study. Clover enhances the grazing value of pasture when grown with grasses. PMID- 11384044 TI - Blood amino acids and milk composition from cows fed soybean meal, fish meal, or both. AB - Twelve multiparous Holstein cows at 48 +/- 8 d in milk were used in a 4 x 4 Latin square with 21-d periods to determine the effect on feed intake, milk yield, milk composition, and blood amino acids when soybean meal was replaced with fish meal. Fish meal substituted for soybean meal on an isonitrogenous basis at 0, 25, 50, and 100% of supplemental protein. Total mixed diets were (dry matter basis) 25% corn silage, 25% alfalfa hay, and 50% concentrate mix. Intake of dry matter (27.9, 27.8, 26.1, and 25.8 kg/d for diets 1 to 4, respectively) was similar for all diets. Milk yield (37.5, 37.8, 37.2, and 37.7 kg/d) was not affected by diets. Milk protein percentages (3.23, 3.24, 3.31, and 3.35) increased with 100% fish meal supplementation and tended to be higher, with 50% fish meal supplementation compared with 100% soybean meal diet. Milk fat percentages (3.18, 2.99, 3.04, and 2.87) and yields were lower with the 100% fish meal than with the 100% soybean meal diet. Molar proportions of ruminal volatile fatty acids and ammonia were not greatly affected by diet. Fish meal supplementation slightly improved Met status, as shown when blood amino acid data were evaluated. Both extraction efficiency and transfer efficiency of amino acids from the blood by the mammary gland indicated that Met, Lys, and Phe were the most limiting amino acids in all diets. Replacing as much as 50 or 100% of dietary soybean meal with fish meal may improve the amino acid balance and increase the protein content in milk; however, feeding 100% fish meal will likely decrease milk fat percentages. PMID- 11384045 TI - Effects of abomasal infusion of long-chain fatty acids on intake, feeding behavior and milk production in dairy cows. AB - Fat is often fed to dairy cows to increase the energy concentration of their diet; however, feeding fat often reduces dry matter intake (DMI), which limits its impact on metabolizable energy (ME) intake. To investigate the effects of postruminal fat infusion on intake, feeding behavior, and milk production of dairy cows at two stages of lactation (55 and 111 d postpartum), six Holstein x British Friesian cows were infused into the abomasum, with a mixture of rapeseed and sunflower oils supplying predominantly unsaturated long-chain fatty acids (LCFA). Dry matter intake was significantly depressed by oil infusion, but estimated ME intake was unchanged, and thus there was no effect of oil infusion on milk yield. There was no effect of stage of lactation on the DM or ME intake response to oil infusion. Milk fat concentration was increased by oil infusion in mid-lactation but not in early lactation, suggesting that the infused LCFA were utilized differently in early compared with midlactation. Cows spent an average of 654 min idling, 462 min ruminating, and 248 min eating during the last 22.8 h of each infusion. There were no significant effects of oil infusion or stage of lactation on the total time spent engaged in these activities. An assessment of the circadian pattern of feeding behavior suggested that the depression in DMI in response to oil infusion occurred after the 1630 and 2230 h feeding times. This may reflect differences in mechanisms regulating feed intake behavior and appetite during the day. Comparison of the results of the present study with the results of other trials involving postruminal fat infusion suggests that polyunsaturated nonesterified fatty acids have the most potent effect on DMI intake. PMID- 11384046 TI - Milk composition responses to unilateral arterial infusion of complete and histidine-lacking amino acid mixtures to the mammary glands of cows. AB - To evaluate a close mammary infusion technique for the study of milk protein responses to blood amino acid profile, five early-lactation, multiparous Holstein cows were surgically fitted with catheters in both external iliac arteries. Animals were infused into one arterial catheter with five different solutions on 5 consecutive days in a Latin square design. Infusions began at 0800 h and continued until 1800 h. The five infusates were a 3% saline control, 15 g/h of complete amino acid mix, 15 g/h of imbalanced amino acid mix (minus His), 30 g/h of complete amino acid mix, and 30 g/h of imbalanced amino acid mix (minus His). Cows were fed a total mixed ration twice daily containing 16% crude protein and 1.7 Mcal/kg of net energy for lactation. Infusion of the complete amino acid mix elevated amino acid concentrations in arterial plasma two- to threefold but caused only a small dose-dependent increase in milk protein content and yield. Fat percentage in milk was decreased from 4.08 to 3.35% by the complete amino acid infusions so that the protein:fat ratio climbed from 0.76 on the control to 0.99 with 30 g/h of amino acid. Removal of His from the infusate caused plasma His concentrations to drop but had no effect on any other circulating amino acids. Milk composition was restored to control levels by removal of the single amino acid. A short-term circulating amino acid imbalance depresses milk protein percentage and increases milk fat content in dairy cows. PMID- 11384047 TI - Body composition and estimated tissue energy balance in Jersey and Holstein cows during early lactation. AB - The rate and extent of estimated energy mobilization and the relationship between fat depth at the rib and thurl and body condition score (BCS) were investigated in Jersey and Holstein cows in early lactation. Twenty-six cows were paired by breed, parity, and calving date, and were individually fed a total mixed ration ad libitum from parturition through 120 d in milk. Feed intake and milk production were measured daily; body weight (BW), BCS, subcutaneous fat depth, milk composition, and concentration of plasma nonesterified fatty acids were measured every 2 wk. Estimated tissue energy balance (TEB) was calculated using 1989 NRC equations. Net energy intake was greater in early lactation for Holsteins compared with Jerseys, 37.8 and 28.2 Mcal/d, respectively. Milk energy was greater for Holsteins relative to Jerseys, 30.5 versus 21.2 Mcal/d. Fat depth and BCS did not differ between breeds. A positive relationship existed between fat depth and BCS for Jerseys; however, there was no significant relationship for Holsteins. The best-fit regression model for predicting TEB for Holsteins and Jerseys in early lactation included week of lactation, milk composition, and BCS. Jerseys remained in negative TEB for a shorter period of time relative to Holsteins. The TEB nadir was -6.19 and -12.9 Mcal/d, for Jerseys and Holsteins, respectively. Expressed as a proportion of metabolic BW (BW(0.75)), net energy intake did not differ between breeds, yet milk energy and estimated tissue energy loss were greater for Holsteins compared with Jerseys. PMID- 11384048 TI - Nitrogen and phosphorus partitioning in lactating Holstein cows fed different sources of dietary protein and phosphorus. AB - To evaluate dietary N and P partitioning, 36 Holstein cows grouped by parity were assigned at calving to diets supplemented with soybean meal (S) or a combination of S and blood meal (B). Diets S and B were formulated to contain 16.2% CP and 0.35% P using mono- and dicalcium phosphate (PM) or wheat bran (WB) as the supplemental source of P. Actual dietary P contents were 0.38, 0.36, 0.34, and 0.34% for SPM, BPM, SWB, and BWB. Two-day total collections of feces, urine, and milk were conducted between 30 and 45 d in milk (DIM), then all cows were fed a control diet until 120 DIM. Between 120 and 150 DIM, cows were again fed the diet assigned at calving, then 2-d total collections of feces, urine, and milk were conducted. Milk production was similar for cows fed diets containing WB (SWB or BWB) when compared with cows fed PM. However, DMI tended to be lower, and P intake and total P excretion were lower in response to WB (20.7 kg/d, 71.9 g/d, and 40.3 g/d) compared with cows fed PM (23.0, 86.7, and 46.8 g/d). Apparent digestibility of dietary P did not differ due to source of supplemental P, averaging 45% across diets. The lower P intake by cows fed WB resulted in lower absorbed P and lower retained P (32.2 and 7.5 g/d) compared with those fed PM (40.6 and 13.4 g/d). Apparent N digestibility, urinary N, and N retention were not affected by P source. Blood meal decreased apparent N digestibility and absorbed N, and also decreased P retention compared with S. In later lactation, cows retained proportionately more absorbed N and P in body tissue and secreted less in milk than they did in early lactation. Results indicated the organic source of P (phytate-P) in WB can be used to provide a substantial portion of the P needed in dairy cattle diets after peak lactation, but the amount of WB in the diet during early lactation should be limited to prevent suppression of DMI and P retention. PMID- 11384049 TI - Effect of chromium supplementation on production and metabolic parameters in periparturient dairy cows. AB - The effect of supplemental chromium as chromium-methionine (Cr-Met) on production and metabolic parameters was investigated in 48 cows from 28 d before expected calving date through 28 d of lactation. Average body weight (BW) was 730 +/- 61 kg before treatment. Treatments were supplementation of 0, 0.03, 0.06, and 0.12 mg of Cr as Cr-Met/kg of BW(0.75). Dry matter intake increased linearly and quadratically during the prepartum and postpartum periods, respectively, and body condition score loss decreased linearly during the postpartum period with increasing Cr-Met. Increasing Cr-Met supplementation caused quadratic increases in milk, fat, and lactose yields. Treatments did not affect concentrations of blood metabolites and liver triglyceride. Serum insulin concentration and molar ratio of insulin to glucose for cows receiving Cr-Met were lower than for cows not receiving Cr-Met, but they increased quadratically with increasing Cr-Met. Glucose tolerance tests (GTT) were conducted on d 10 prepartum and d 28 postpartum. Chromium-methionine supplementation attenuated insulin sensitivity prepartum and enhanced glucose tolerance postpartum, but not prepartum. Basal insulin concentrations for cows receiving Cr-Met were higher than for cows not receiving Cr-Met during the prepartum GTT. During the postpartum GTT, peak glucose concentration and clearance rate decreased and half-life (t(1/2)) and time to reach basal concentration (T) were prolonged quadratically by increasing Cr-Met. Additionally, peak insulin concentration, area under the curve, and molar ratio of insulin to glucose were higher for cows not receiving Cr-Met than for cows receiving Cr-Met and t(1/2) and T were shortened quadratically by increasing Cr-Met. PMID- 11384050 TI - Responses of milk fat composition to dietary fat or nonstructural carbohydrates in Holstein and Jersey cows. AB - Milk fat from Jersey cows contains less oleic acid (cis-C18:1) and more short- and medium-chain fatty acids than does milk fat from Holstein cows. The objective of this experiment was to determine responses in milk fat composition when Jersey and Holstein cows were fed diets either high (37% of dry matter) or low (27% of dry matter) in content of nonstructural carbohydrates (NSC) and supplemented with either 0 or 2.5% (of dry matter) of a mostly saturated fat source. Four Holstein cows and four Jersey cows were used in a Latin square design with 28-d periods; diets were in a 2 x 2 factorial arrangement. Fat supplementation decreased contents of fatty acids synthesized de novo within the mammary gland and increased contents of C18:0 and cis-C18:1. Low-NSC diets tended to increase C16:0 and to decrease C18:0, cis-C18:1, and C18:3. Despite the differences in fatty acid composition between breeds, both breeds generally responded similarly to dietary treatments. An interaction of breed and fat indicated that the content of cis-C18:1 in milk fat was increased more by supplemental fat in Holsteins than in Jerseys. Interactions of breed x fat and breed x carbohydrate type showed that the ratio of cis-C18:1 to C18:0 decreased when Jerseys were supplemented with fat but increased for Holsteins, and decreased when Jerseys were fed the low-NSC diet but increased when Holsteins were fed low NSC. The data are consistent with the hypothesis (Beaulieu and Palmquist, 1995, J. Dairy Sci. 78:1336-1344) that mammary activity of stearoyl-coenzyme A desaturase is lower in Jerseys than in Holsteins. PMID- 11384051 TI - Computing mating bull fertility from DHI nonreturn data. AB - Animal model methodology was used to compute yearly measures of relative fertility of Holstein AI mating bulls based upon 70-d nonreturn of first breedings as reported to U.S. DHIA from 1988 through 1997. Estimated Relative Conception Rates (ERCR) were computed for bulls with a minimum of 50 first breedings in a single year using variance ratios 45.5 for mating bull, 45.5 for animal genetic effects, and 31 for permanent environment. The model assumed repeatability across lactations of 0.05 and included fixed effects of herd-year month bred and classes of parity, early lactation energy-corrected milk and days open when bred. Estimates of fertility were greater for breedings to cows that were young, had low early lactation production, and were in late stages of lactation. ERCR were expressed as difference in nonreturn from the average AI mating bull of herdmates. Values ranged from -18 to +13. For ERCR computed from a minimum of 1000 breedings, 90% were within four units of zero. Early ERCR computed from a few breedings in a single year were tested for ability to predict later ERCR computed from a minimum of 1000 different breedings. Early ERCR computed from 300 or more matings accurately predicted later independent ERCR. For yearly estimates each based upon a minimum of 1000 breedings, 8% changed more than three units, and 4% declined more than three units. Correlations between ERCR and predicted transmitting abilities protein and type production index were significant but accounted for little variance. Correlations between ERCR and other traits were not significant. PMID- 11384052 TI - Genetic evaluation of Holstein sires and maternal grandsires in the United States for perinatal survival. AB - Stillbirth, defined as a calf that dies just prior to, during, or within 48 h of parturition, represents a reoccurring concern among breeders of dairy cattle in the United States. About 11% of parturitions of primiparous Holstein cows result in the death of a calf; 5.7% in multiparous cows. Genetic evaluations can be reported as perinatal survival to 48 h to emphasize the positive information about the trait. The purpose of this research was to: 1) estimate genetic parameters by restricted maximum likelihood for perinatal survival rates; 2) characterize the genetic evaluation of sires for the perinatal survival of their progeny and maternal grandsires for the perinatal survival of their daughters progeny; and 3) estimate genetic trends from 1984 to 1994. Data (n = 666,339) were from the National Association of Animal Breeders calving ease database. Over 600 new young sires were available each year. The binomial response variable, 1 = alive, 0 = stillborn within 48 h of parturition was analyzed by using a sire maternal grandsire linear mixed model. The model included fixed effects for sex of calf, dystocia, and season of birth, and gestation length as a covariate; correlated random effects of sire and maternal grandsire; and uncorrelated random effects of herd-years. Parturitions of primiparous and multiparous cows were analyzed separately. In primiparous cows, heritability estimates were 1.1 and 2.2% for sire of the calf and maternal grandsire, respectively. The genetic correlation between sire and maternal grandsire predicted transmitting ability (PTA) for perinatal survival, was 0.31; simple product moment correlations among sire-MGS PTA were 0.43 and 0.46 for primiparous and multiparous cows, respectively. The PTA for sire of the calf ranged from -2.9 (lower survival) to 2.8% (higher survival). Mean PTA from 1984 to 1994 was quite variable from year to year. Evidence showed a slightly negative, but nonsignificant, genetic trend in perinatal survival (-0.04% per year for sires and -0.02% per year for maternal grandsires). Estimates of genetic parameters and genetic trends for data from multiparous cows are also reported. Correlations among PTA for perinatal survival, milk yield, and calving ease are given. PMID- 11384053 TI - Estimates of genetic parameters for daily somatic cell count of Australian dairy cattle. AB - Genetic parameters for daily somatic cell counts (SCC) of the first three parities were estimated for Australian Dairy Cattle. Most of the data analyses were carried out with a sire random regression model. The estimates were compared with those from conventional ten-trait analyses and animal models. In the first parity estimates of heritabilities (h2) were low (0.04 to 0.05) at the beginning of the lactation and higher (0.11 to 0.13) at the end. The average h2 estimated from random regression sire model, random regression animal model and conventional multitrait sire model were 0.09, 0.09, and 0.08, respectively, in the first lactation. The average h2 were 0.09 and 0.11 in the second and third parities, respectively. Genetic correlations between daily log(e) SCC within parity were high for adjacent tests (nearly 1) and low (as low as 0.30) between the beginning and the end of the lactation. Generally, the genetic correlations between parities depend on how far apart they are and on whether they are on the same day in any two parities. Across parities, on average, genetic correlations between parities 1 and 3 were the lowest and those between 1 and 2 intermediate, while those between 2 and 3 were the highest. The estimated environmental correlations were lower than the genetic correlations, but the trends were generally similar. Differences in genetic parameter estimates due to model were small, except for some genetic correlations. The high residual error variances, the low h2, and the inconsistency in genetic correlations that were observed particularly at the beginning of the first lactation suggest that log(e) SCC early in the first lactation may be related to a spike in SCC as result of infection and (or) onset of lactation while SCC later in lactation represents a sustained response to infection. Accounting for the variation in heritabilities and correlations should improve the accuracy of genetic evaluations for SCC based on test day records. PMID- 11384054 TI - Genetic analysis of body condition score of lactating Dutch Holstein and Red-and White heifers. AB - The aim of this study was to estimate phenotypic and genetic parameters for body condition scores (BCS) from the Dutch type classification system. Data included 108,809 Holstein (H) and 26,208 Red-and-White (R) heifers from 9701 herds that were scored once during lactation on a 1 to 9 scale (1 = emaciated and 9 = obese). Mean BCS for H and R data were 4.50 and 4.94, respectively. The BCS decreased as the percentage of Holstein genes increased. For both breeds, BCS after calving was about 5.6 and BCS was lowest around wk 11. For H heifers, mean BCS at drying off was about 0.8 lower than BCS at calving, whereas for R heifers BCS was at about the same level as at calving. Variance components were estimated using an animal model including the effects of herd x visit, classifier, age at calving, DIM, and genetic group. The random herd x visit effect explained about 10 to 15% of the phenotypic variation. Heritabilities ranged from 0.24 to 0.38, depending on breed and lactation period. Genetic correlations between BCS observations in bimonthly lactation periods were close to unity, especially for H. It was concluded that BCS data collected by type classifiers can well be used for genetic evaluation and that genetic variation between animals for BCS-change patterns is a small component of the overall variation in BCS. PMID- 11384055 TI - Distribution of bovine milk sialoglycoconjugates during lactation. AB - The sialoglycoconjugate content of human milk has been extensively studied. However, little attention has been paid to the changes occurring in these compounds in bovine milk during lactation. Since sialoglycoconjugates are very abundant in milk from the early stages of lactation, they have been suggested to be important for the nutrition of the newborn during the first months of life. The distribution of sialoglycoconjugates (expressed as glycoconjugate-bound sialic acid) from four different stages of lactation (colostrum, transitional, mature, and late-lactation milks) was investigated in four Spanish-Brown cows. All the fractions studied (total sialic acids, glycoproteins, oligosaccharides, casein, and gangliosides) showed a similar trend. We found the highest values in the colostrum, these decreasing in transitional and mature milks and increasing again in late-lactation milk. We also found a selective change in the relative contents of glycoprotein- and oligosaccharide-bound sialic acids. In mature milk, the latter increased up to 80% (59% in colostrum) and the former decreased to 3.9% (35.3% in colostrum). It would appear that the decrease in oligosaccharide bound sialic acid is compensated by the increase in glycoprotein-bound sialic acid. From these results, it is deduced that newborn infants or calves fed with infant formulas or milk replacers, respectively, should be supplemented with sialoglycoconjugates to approximate the composition of human and cow milk as far as is practicable. PMID- 11384056 TI - Importance of the choice of the collimator for the detection of small lesions in scintimammography: a phantom study. AB - 99mTc methoxyisobutylisonitrile planar scintimammography (SMM) is mostly performed using low-energy high-resolution (LEHR) parallel collimators. We studied whether using a different collimator could improve the detection of small (< 1.5 cm) lesions for which SMM sensitivity is poor. Thirty four breast phantom configurations were considered, either with hot spheres simulating lesions or without any spheres. For each configuration, four planar acquisitions were performed using LEHR, low-energy ultra high-resolution (LEUHR), high-resolution fan-beam (HRFB) and ultra high-resolution fan-beam (UHRFB) collimators. Images corresponding to the 20% and 10% energy windows and to the Jaszczak subtraction were calculated. A database including 156 borderline images was derived. After training, 10 observers scored the images for the presence of a sphere. The performances in sphere detection were studied using receiver operating characteristic (ROC) analysis. For all types of image, the area under the ROC curve was highest with the UHRFB collimator and lowest with either the LEUHR or the HRFB collimator. For the 10% energy window images conventionally used in SMM, the detection sensitivities averaged 91%, 73%, 60% and 55% for the UHRFB, LEHR, HRFB and LEUHR collimators respectively, for the same specificity of 64%. We conclude that detection of small tumours in planar SMM might be significantly improved by using a UHRFB collimator instead of an LEHR collimator. PMID- 11384057 TI - The use of SPAMM to assess spatial distortion due to static field inhomogeneity in dental MRI. AB - In planning placement of dental implants using MRI, a SPAMM (spatial modulation of magnetization) magnetization preparation sequence was incorporated into a spin echo imaging sequence. A phantom was imaged with a ferromagnetic object attached. Spatial distortion due to deviations in Larmor frequency was detected by a deviation of SPAMM lines. Both SPAMM line deviation and interline spacing were found to agree with a deltaB0 map generated from phase images. Imaging of a volunteer with and without typically used metallic implants positioned in a template showed SPAMM line deviations to correlate with expected deviations in vivo. SPAMM lines showed possible distortion due to chemical shift in the bone marrow and the presence of titanium implants to be insignificant. SPAMM may thus be used to provide a qualitative estimate of the accuracy of the MRI image when planning dental implants. PMID- 11384058 TI - Modelling the dosimetric consequences of organ motion at CT imaging on radiotherapy treatment planning. AB - Treatment planning algorithms usually assume that the correct or at least the mean organ position is derived from the CT imaging procedure, and that this position is reproduced throughout the treatment. In reality a mobile organ is unlikely to be in its exact mean position at the time of imaging, causing the treatment to be planned with an organ off-set from its assumed mean position. This introduces an extra 'CT uncertainty' into the treatment. A Monte Carlo (MC) model is used to simulate organ translations at imaging and evaluate the effect of this uncertainty (above the treatment delivery uncertainties) on the dose distribution. An underdose by 4 Gy in a 60 Gy treatment is calculated in the penumbral region of a single-field dose distribution as a result of the CT uncertainty. The effect is reduced to less then 0.5 Gy when the organ position at planning is derived as the average from multiple pretreatment CT scans. It is shown that a convolution method can be applied to predict the effect of CT uncertainty on the dose distribution for a patient population. Additionally, a variation kernel for a convolution method is derived that incorporates uncertainty at both imaging and treatment. PMID- 11384059 TI - A wide dynamic range, high-spatial-resolution scanning system for radiochromic dye films. AB - This paper describes the evaluation of an inexpensive, commercially available 35 mm transparency slide scanner as a potential alternative scanning device for GafChromic HD-810 radiochromic dye film. Besides its low cost, the principal advantages of this type of scanner are high spatial resolution and high speed (a typical scan taking less than 1 min). With broad-band illumination the useful dose range using grey-scale imaging of GafChromic HD-810 is limited to about 50 800 Gy. By using the colour-scale imaging capability of the scanner we have been able to achieve a significant extension covering a similar range (15-2000 Gy) to that attainable using monochromatic illumination. The short-term reproducibility of the system is good, with a coefficient of variation of doses estimated from repeat scanning of uniformly exposed calibration films of less than 2%. Long-term stability is ensured by the scanning of a manufacturer-supplied test slide. The slide scanner system has been used in the determination of depth dose distributions from a model 'hot particle' source containing 106Ru/Rh. GafChromic dye film stacks irradiated by the source were read out on both the slide scanner and a conventional Joyce Loebl MDM6 scanning stage microdensitometer. The overall agreement between the dose estimates provided by the two systems was within 10%. PMID- 11384060 TI - Directional dependence in film dosimetry: radiographic and radiochromic film. AB - The trend towards conformal, intensity modulated radiotherapy treatments has established the need for a true integrating dosimeter. In traditional radiotherapy, radiographic film dosimetry is commonly used. The accuracy and reproducibility of film optical density as an indicator of dose is influenced by several variables, including the chemical processing conditions. As a result radiochromic film, with all the advantages of radiographic film but without the need for chemical processing, has increased in popularity, although the low-dose sensitivity of radiochromic film does remain a disadvantage for some experiments. Several studies have investigated the reproducibility of radiochromic film results, but none have specifically addressed the well-known directional dependence seen with traditional radiographic film. In this study, the directional dependence of radiographic (Kodak X-omat V) and radiochromic (Gafchromic) films were measured. It was found that both films over responded when exposed parallel to the central axis of the beam as opposed to perpendicular exposure. An attempt is made to explain the reason for the responses of both films in terms of spectral effects and the air gap between the phantom segments. Although radiographic film exposed parallel rather than perpendicular to the central axis of the beam exhibits a measured difference in film response at depth, this over response does not occur when the extent of the film is restricted to a small region at the centre of the phantom (in this case an air gap is not introduced across the phantom). This suggests that it is the air gap rather than the orientation of the film that is the cause of the over response. Furthermore, when film occupies a slice through the entire phantom an over response occurs for both radiographic and radiochromic film, indicating that spectral effects are not the cause. PMID- 11384061 TI - A computational study into the use of polyacrylamide gel and A-150 plastic as brain tissue substitutes for boron neutron capture therapy. AB - A precise evaluation of the dosimetric performance of epithermal neutron beams designed for boron neutron capture theory of brain tumours requires the use of a phantom material that closely matches brain tissue. The aim of this study was to investigate how well polyacrylamide gel (or PAG) and A- 150 plastic performed as substitutes for brain tissue compared with standard phantom materials such as water and polymethyl-methacrylate (or PMMA). Thermal neutron fluence, photon dose and epithermal neutron dose distributions were calculated for the epithermal neutron beam available at the University of Birmingham. The results presented in this paper show that the PAG provides a good simulation of radiation transport in the brain with differences from the real brain of +9.4%, - 10.8% and +5.1% at a depth of 50 mm for thermal neutron fluence, gamma dose and epithermal neutron dose distributions respectively. The polyacrylamide gel presented is therefore a promising substitute for brain tissue that can, as a dosimeter, provide a three dimensional map of the absorbed dose delivered by the epithermal neutron beam. However, this study does not investigate the agreement between doses derived from magnetic resonance and physical doses for such gels. A- 150 plastic was shown to be a better substitute for brain tissue than PMMA, with differences from brain of -1.9%, -12.4% and - 13.2% at a depth of 50 mm for thermal neutron fluence, gamma dose and epithermal neutron dose distributions respectively, against +21.1%, 16.2% and +19.2% for PMMA. A-150 plastic should therefore be the material of choice for solid phantoms. PMID- 11384062 TI - A multiple source model for 6 MV photon beam dose calculations using Monte Carlo. AB - A multiple source model (MSM) for the 6 MV beam of a Varian Clinac 2300 C/D was developed by simulating radiation transport through the accelerator head for a set of square fields using the GEANT Monte Carlo (MC) code. The corresponding phase space (PS) data enabled the characterization of 12 sources representing the main components of the beam defining system. By parametrizing the source characteristics and by evaluating the dependence of the parameters on field size, it was possible to extend the validity of the model to arbitrary rectangular fields which include the central 3 x 3 cm2 field without additional precalculated PS data. Finally, a sampling procedure was developed in order to reproduce the PS data. To validate the MSM, the fluence, energy fluence and mean energy distributions determined from the original and the reproduced PS data were compared and showed very good agreement. In addition, the MC calculated primary energy spectrum was verified by an energy spectrum derived from transmission measurements. Comparisons of MC calculated depth dose curves and profiles, using original and PS data reproduced by the MSM, agree within 1% and 1 mm. Deviations from measured dose distributions are within 1.5% and 1 mm. However, the real beam leads to some larger deviations outside the geometrical beam area for large fields. Calculated output factors in 10 cm water depth agree within 1.5% with experimentally determined data. In conclusion, the MSM produces accurate PS data for MC photon dose calculations for the rectangular fields specified. PMID- 11384063 TI - Derivation of electron and photon energy spectra from electron beam central axis depth dose curves. AB - A method for deriving the electron and photon energy spectra from electron beam central axis percentage depth dose (PDD) curves has been investigated. The PDD curves of 6, 12 and 20 MeV electron beams obtained from the Monte Carlo full phase space simulations of the Varian linear accelerator treatment head have been used to test the method. We have employed a 'random creep' algorithm to determine the energy spectra of electrons and photons in a clinical electron beam. The fitted electron and photon energy spectra have been compared with the corresponding spectra obtained from the Monte Carlo full phase space simulations. Our fitted energy spectra are in good agreement with the Monte Carlo simulated spectra in terms of peak location, peak width, amplitude and smoothness of the spectrum. In addition, the derived depth dose curves of head-generated photons agree well in both shape and amplitude with those calculated using the full phase space data. The central axis depth dose curves and dose profiles at various depths have been compared using an automated electron beam commissioning procedure. The comparison has demonstrated that our method is capable of deriving the energy spectra for the Varian accelerator electron beams investigated. We have implemented this method in the electron beam commissioning procedure for Monte Carlo electron beam dose calculations. PMID- 11384064 TI - A simulation of the effects of set-up error and changes in breast volume on conventional and intensity-modulated treatments in breast radiotherapy. AB - The effect of interfractional patient movement on dosimetry has been investigated for breast radiotherapy. Errors in patient set-up and changes in breast volume were simulated individually to determine how each contributes to the total dosimetric error. Two treatment techniques were investigated: a conventional treatment and an intensity-modulated treatment delivered using compensators. Six patients were investigated and anterior-posterior (AP) and superior-inferior (SI) displacements were simulated by displacing the isocentre in both directions by 2, 5 and 10 mm. A model of the breast was developed from the six patients to simulate changes in breast volume. In this model, the breast was described as a set of semi-ellipses. The volume of the breast was changed by varying the magnitude of the semi-major and semi-minor axes. Anisotropic changes in breast volume were also investigated. The dosimetric error was evaluated for each dose plan by calculating the volume outside the 95-105% dose range resulting from the simulations. A number of parameters describing the size and shape of the breast were also investigated to determine whether a susceptibility of outline sets to interfractional patient movement could be predicted. A parameter describing the increase in the breast volume outside the 95-105% dose range was calculated for AP a PMID- 11384065 TI - A practical approach to inverse planning for high-precision dose escalated conformal prostate radiotherapy. AB - The problem of choosing the best gantry angles and beam weights for dose escalated conformal prostate treatment planning is formulated using a mixed integer linear programming approach, to account for tumour dose homogeneity and dose-volume constraints. The formulation allows the number of beams to be restricted and for some of the beams to be compulsory. The present planning algorithm interfaces with and utilizes the three-dimensional planning capabilities of a commercial treatment planning system. A case study is illustrated, which represents a particularly challenging planning problem due to a large planning target volume and an unusually small bladder. Treatment plans with different numbers of beams are generated to compare with each other and with the standard six-field plan. Significant improvement is shown in the reduction of hot regions within the femoral heads and rectal wall, while not unduly compromising homogeneity constraints for the tumour. PMID- 11384066 TI - Anisotropy function for 192Ir low-dose-rate brachytherapy sources: an EGS4 Monte Carlo study. AB - The anisotropy function of low-dose-rate 192Ir interstitial brachytherapy sources was studied. Absolute dose rates around 192Ir seeds with stainless steel or platinum cladding and a platinum covered wire have been estimated using the EGS4 Monte Carlo simulation system with a very well tested user code. Our results were compared with available experimental data. Excellent agreement between calculated and measured anisotropy function was found for stainless steel clad 192Ir sources, except along the longitudinal axis of the sources. Dosimetry data for the platinum covered seed and 3 mm long wire with platinum cladding as well as for the stainless steel clad 192Ir source are presented in TG43 format. The influence of phantom dimensions on the anisotropy function was found to be non negligible over 7 cm, enhancing the anisotropy function by 1-2%. Our results have estimated statistical uncertainties below 1% at 1 sigma level up to 10 cm excluding the longitudinal axis where statistical uncertainties below 3% up to 10 cm are observed. PMID- 11384067 TI - Generalization of a model of tissue response to radiation based on the idea of functional subunits and binomial statistics. AB - This work investigates the existing biological models describing the response of tumours and normal tissues to radiation, with the purpose of developing a general biological model of the response of tissue to radiation. Two different types of normal tissue behaviour have been postulated with respect to its response to radiation, namely critical element and critical volume behaviour. Based on the idea that an organ is composed of functional subunits, models have been developed describing these behaviours. However, these models describe the response of an individual, a particular patient or experimental animal, while the clinically or experimentally observed quantity is the population response. There is a need to extend the models to address the population response, based on the ideas we have about the individual response. We have attempted here to summarize and unify the existing individual models. Finally, the population models are investigated by fitting to pseudoexperimental sets of data and comparing them with each other in terms of goodness-of-fit and in terms of their power to recover the values of the population parameters. PMID- 11384068 TI - The inclusion of capillary distribution in the adiabatic tissue homogeneity model of blood flow. AB - We have developed a non-invasive imaging tracer kinetic model for blood flow which takes into account the distribution of capillaries in tissue. Each individual capillary is assumed to follow the adiabatic tissue homogeneity model. The main strength of our new model is in its ability to quantify the functional distribution of capillaries by the standard deviation in the time taken by blood to pass through the tissue. We have applied our model to the human prostate and have tested two different types of distribution functions. Both distribution functions yielded very similar predictions for the various model parameters, and in particular for the standard deviation in transit time. Our motivation for developing this model is the fact that the capillary distribution in cancerous tissue is drastically different from in normal tissue. We believe that there is great potential for our model to be used as a prognostic tool in cancer treatment. For example, an accurate knowledge of the distribution in transit times might result in an accurate estimate of the degree of tumour hypoxia, which is crucial to the success of radiation therapy. PMID- 11384069 TI - Quasistatic zooming of FDTD E-field computations: the impact of down-scaling techniques. AB - Due to current computer limitations, regional hyperthermia treatment planning (HTP) is practically limited to a resolution of 1 cm, whereas a millimetre resolution is desired. Using the centimetre resolution E-field distribution, computed with, for example, the finite-difference time-domain (FDTD) method and the millimetre resolution patient anatomy it is possible to obtain a millimetre resolution SAR distribution in a volume of interest (VOI) by means of quasistatic zooming. To compute the required low-resolution E-field distribution, a low resolution dielectric geometry is needed which is constructed by down-scaling the millimetre resolution dielectric geometry. In this study we have investigated which down-scaling technique results in a dielectric geometry that yields the best low-resolution E-field distribution as input for quasistatic zooming. A segmented 2 mm resolution CT data set of a patient has been down-scaled to 1 cm resolution using three different techniques: 'winner-takes-all', 'volumetric averaging' and 'anisotropic volumetric averaging'. The E-field distributions computed for those low-resolution dielectric geometries have been used as input for quasistatic zooming. The resulting zoomed-resolution SAR distributions were compared with a reference: the 2 mm resolution SAR distribution computed with the FDTD method. The E-field distribution for both a simple phantom and the complex partial patient geometry down-scaled using 'anisotropic volumetric averaging' resulted in zoomed-resolution SAR distributions that best approximate the corresponding high-resolution SAR distribution (correlation 97, 96% and absolute averaged difference 6, 14% respectively). PMID- 11384070 TI - 4D maximum a posteriori reconstruction in dynamic SPECT using a compartmental model-based prior. AB - A 4D ordered-subsets maximum a posteriori (OSMAP) algorithm for dynamic SPECT is described which uses a temporal prior that constrains each voxel's behaviour in time to conform to a compartmental model. No a priori limitations on kinetic parameters are applied; rather, the parameter estimates evolve as the algorithm iterates to a solution. The estimated parameters and time-activity curves are used within the reconstruction algorithm to model changes in the activity distribution as the camera rotates, avoiding artefacts due to inconsistencies of data between projection views. This potentially allows for fewer, longer-duration scans to be used and may have implications for noise reduction. The algorithm was evaluated qualitatively using dynamic 99mTc-teboroxime SPECT scans in two patients, and quantitatively using a series of simulated phantom experiments. The OSMAP algorithm resulted in images with better myocardial uniformity and definition, gave time-activity curves with reduced noise variations, and provided wash-in parameter estimates with better accuracy and lower statistical uncertainty than those obtained from conventional ordered-subsets expectation maximization (OSEM) processing followed by compartmental modelling. The new algorithm effectively removed the bias in k21 estimates due to inconsistent projections for sampling schedules as slow as 60 s per timeframe, but no improvement in wash-out parameter estimates was observed in this work. The proposed dynamic OSMAP algorithm provides a flexible framework which may benefit a variety of dynamic tomographic imaging applications. PMID- 11384071 TI - A comparison of mammography spectral measurements with spectra produced using several different mathematical models. AB - Due to the relatively complex nature of spectral measurements from x-ray machines, many researchers use mathematical models to simulate the spectra they need. However, there is concern over their accuracy, and hence the impact that their accuracy may have, on subsequent calculations that rely upon the spectra modelled. With this in mind spectral measurements have been performed on a mammography machine and a comparison with spectra calculated using several different models is presented. Several different techniques have been investigated in the spectral measurements to allow for pulse pileup and other effects of high count rate. Comparison with half value layer (HVL) measurements shows that the use of a gating signal in conjunction with the air-free path provides accurate results without the need for a pinhole collimator. Comparison of the measured spectra with those calculated using different models proposed in the literature suggests that accurate results can be produced by all models, but only if the user attempts to match the calculated HVL of the modelled spectrum with the physically measured HVL. If this is not done the modelled spectra may be in error. The impact of such an error is demonstrated in calculations of mean glandular dose, which indicate a possible underestimate of the dose by up to 20%. PMID- 11384072 TI - An apparatus for applying strong longitudinal magnetic fields to clinical photon and electron beams. AB - Monte Carlo studies have recently renewed interest in the use of the effect of strong transverse and longitudinal magnetic fields to manipulate the dose characteristics of clinical photon and electron beams. A 3.5 T superconducting solenoidal magnet was used to evaluate the effect of a longitudinal field on both photon and electron beams. This note describes the apparatus and demonstrates some of the effects on the beam trajectory and dose distributions for measurements in a homogeneous phantom. The effects were studied using film in air and in phantoms which fit in the magnet bore. The magnetic field focused and collimated the electron beams. The converging, non-uniform field confined the beam and caused it to converge with increasing depth in the phantom. Due to the field's collecting and focusing effect, the beam flux density increased, leading to increased dose deposition near the magnetic axis, especially near the surface of the phantom. This study illustrates some benefits and challenges associated with the use of non-uniform longitudinal magnetic fields in conjunction with clinical electron and photon beams. PMID- 11384073 TI - Reduction of computational dimensionality in inverse radiotherapy planning using sparse matrix operations. AB - For dynamic multileaf collimator-based intensity modulated radiotherapy in which small beam elements are used to generate continuous modulation, the sheer size of the dose calculation matrix could pose serious computational challenges. In order to circumvent this problem, the dose calculation matrix was reduced to a sparse matrix by truncating the weakly contributing entries below a certain cutoff to zero. Subsequently, the sparse matrix was compressed and matrix indexing vectors were generated to facilitate matrix-vector and matrix-matrix operations used in inverse planning. The application of sparsity permitted the reduction of overall memory requirement by an order of magnitude. In addition, the effect of disregarding the small scatter components on the quality of optimization was investigated by repeating the inverse planning using the dense dose calculation matrix. Comparison of dense and sparse matrix-based plans revealed an insignificant difference in optimization outcome, thus demonstrating the feasibility and usefulness of the sparse method in inverse planning. Furthermore, two additional methods of memory minimization are suggested, namely hexagonal dose sampling and limited normal tissue sampling. PMID- 11384074 TI - Three-dimensional ultrasound imaging. AB - Ultrasound is an inexpensive and widely used imaging modality for the diagnosis and staging of a number of diseases. In the past two decades, it has benefited from major advances in technology and has become an indispensable imaging modality, due to its flexibility and non-invasive character. In the last decade, research investigators and commercial companies have further advanced ultrasound imaging with the development of 3D ultrasound. This new imaging approach is rapidly achieving widespread use with numerous applications. The major reason for the increase in the use of 3D ultrasound is related to the limitations of 2D viewing of 3D anatomy, using conventional ultrasound. This occurs because: (a) Conventional ultrasound images are 2D, yet the anatomy is 3D, hence the diagnostician must integrate multiple images in his mind. This practice is inefficient, and may lead to variability and incorrect diagnoses. (b) The 2D ultrasound image represents a thin plane at some arbitrary angle in the body. It is difficult to localize the image plane and reproduce it at a later time for follow-up studies. In this review article we describe how 3D ultrasound imaging overcomes these limitations. Specifically, we describe the developments of a number of 3D ultrasound imaging systems using mechanical, free-hand and 2D array scanning techniques. Reconstruction and viewing methods of the 3D images are described with specific examples. Since 3D ultrasound is used to quantify the volume of organs and pathology, the sources of errors in the reconstruction techniques as well as formulae relating design specification to geometric errors are provided. Finally, methods to measure organ volume from the 3D ultrasound images and sources of errors are described. PMID- 11384075 TI - Predictive risk factors for pulmonary oxygen transfer in patients undergoing coronary artery bypass grafting. AB - The ratio of arterial oxygen tension to inspired oxygen fraction (PaO2/FiO2) is a useful indicator for weaning patients from mechanical ventilation and a reliable predictor of pulmonary dysfunction after cardiac surgery. The aim of this study was to elucidate the patient characteristics and variables that affect the PaO2/FiO2 ratio. Between 1994-1998, 167 patients who underwent coronary artery bypass grafting (CABG) were examined retrospectively. Spearman's correlation coefficients were calculated between the PaO2/FiO2 ratio and intubation period, and length of ICU stay. Patients were then divided into two groups with a PaO2/FiO2 ratio < 350 and PaO2/FiO2 ratio > or = 350. Univariate analysis of the putative risk factors was performed. A logistic regression model was developed to evaluate factors that would influence the PaO2/FiO2 ratio. A significant correlation was observed between the PaO2/FiO2 ratio and intubation period, and length of ICU stay. Univariate predictors of a PaO2/FiO2 ratio < 350 were low body weight, low preoperative PaO2 long operation time, high FiO2, low postoperative PaO2 history of smoking, hypertension and opening of pleura (p < 0.05). Excellent prediction was found with a model consisting of preoperative PaO2 and hypertension. CONCLUSION: The results of this study suggest that patients with a low preoperative PaO2 or hypertension may need more careful peri- and postoperative management since these factors are closely associated with the PaO2/FiO2 ratio. PMID- 11384076 TI - Coronary artery bypass grafting with cardiopulmonary bypass is safe and contributes to favorable long-term results. AB - The aim of this study was to determine the indication for minimally invasive direct coronary artery surgery based on the operative and long-term results of conventional coronary artery bypass grafting with cardiopulmonary bypass. Operative results: The subjects included 505 patients who underwent isolated elective coronary artery bypass grafting with cardiopulmonary bypass from January 1995 through August 1999. The mean age at the time of surgery was 61.9 and the mean number of grafts per patient was 2.6. Long-term results: From January 1984 to December 1995, a total of 907 patients received coronary artery bypass grafting with cardiopulmonary bypass using the internal thoracic artery to the left anterior descending artery with or without saphenous vein grafts to other coronary arteries. The rates of complete and incomplete revascularization were 69.3% (n = 629) and 30.7% (n = 278), respectively. Mean follow-up was 5.95+/-3.0 years. The operative results revealed low output syndrome occurred in 14 patients (2.8%), perioperative myocardial infarction with the appearance of new Q-waves in 5 (1.0%), renal failure requiring transient dialysis in 16 (3.2%), stroke with persistent sequelae in 5 (1.0%), and mediastinitis in 5 (1.0%). Two patients (0.4%) died in the hospital. The long-term results for the 907 patients revealed the 10-year actuarial survival, 10-year cardiac death free, and 10-year cardiac event free rates were 85.7%, 94.1%, and 77.3%, respectively. The 10-year survival rates was 88.4% among patients receiving complete revascularization and 79.3% among those receiving incomplete revascularization (p = 0.0334). The 10-year cardiac death free rate among patients undergoing complete revascularization was 96.3% and 88.7% among those receiving incomplete revascularization (p = 0.0016). The 10-year cardiac event free rates were 82.3% and 67.9%) among patients undergoing complete and incomplete revascularization, respectively (p = 0.0118). In view of the favorable operative and long-term results of conventional coronary artery bypass grafting, especially complete revascularization, we conclude that minimally invasive direct coronary artery grafting is an appropriate treatment for multi-vessel disease in carefully selected patients at a high risk for stroke and major comorbidities due to old age. PMID- 11384077 TI - Diurnal variations of post-exercise parasympathetic nervous reactivation in different chronotypes. AB - The purpose of this study was to investigate the diurnal variation and chronotype differences, i.e., in morning-types and evening-types, in post-exercise vagal reactivation. Twelve healthy male college students who were classified as morning type (6) and evening-type (6), based on responses to a questionnaire, participated in this study. Postexercise vagal reactivation was assessed as the time constant of the beat-by-beat heart rate decrease for the first 30 sec after exercise (T30) at an intensity lower than the ventilatory threshold. The subjects performed 3-min cycle ergometer exercise at an intensity corresponding to 80% of the ventilatory threshold after a 1 min warm-up exercise in the morning (7:00 - 8:00) and evening (17:00 - 18:00) to obtain the T30. A significant interaction (chronotype-by-time) effect was found for T30. The morning value of the T30 in evening-type subjects was significantly larger than their evening value and the morning value in morning-type subjects. There was no significant interaction effect for heart rate and oxygen uptake during exercise. These results suggest that diurnal variation in post-exercise vagal reactivation is different between morning-type and evening-type, and post-exercise vagal reactivation in evening type individuals is sluggish in the morning. PMID- 11384078 TI - Quantitative analysis of hypertrophy in cardiac chambers in cyanotic tetralogy of Fallot. AB - Although early total corrective repair for cyanotic tetralogy of Fallot is now safely performed at many institutions, long-term complications after surgical repair have been demonstrated. Therefore, the optimal procedure and timing for surgical treatment remain controxersial. In the present study, we conducted a quantitative analysis of the hypertrophy of all four chambers of 87 autopsied hearts of cyanotic tetralogy of Fallot and 71 normal control hearts utilizing the myocardial mass index, and evaluated the progression of lesions with advancing age. In cyanotic tetralogy of Fallot, hypertrophy of the right ventricle progresses immediately after birth, with that of the right atrium developing soon after. The left side of the heart is normal or slightly atrophied which could be corrected by sufficient palliative intervention or total corrective repair. The growth curves of both ventricles were parallel to those of normal hearts for the period studied. Pulmonary atresia, palliative operation, and total corrective repair have been shown to have some influence on the morphological characteristics of hearts of cyanotic tetralogy of Fallot. PMID- 11384079 TI - Comparison of the first dose response of fosinopril and captopril in congestive heart failure: a randomized, double-blind, placebo controlled study. AB - The purpose of this study was to compare the safety and tolerability of recommended initial doses of fosinopril (FOS) with those of captopril (CAP), in diuretic-treated, salt depleted "high risk" patients with congestive heart failure. Thirty patients were randomized in a double blind fashion to receive a single dose of either FOS 10 mg, CAP 6.25 mg or placebo. CAP produced a significant early and brief fall in BP, while the first-dose hypotensive response with FOS did not differ significantly from placebo. Baseline plasma angiotensin converting enzyme (ACE) activity was similar in all groups. Only CAP showed an acute and significant fall in plasma ACE activity, whereas FOS and placebo did not change ACE activity. There was no correlation between mean arterial pressure or percentile change in mean arterial pressure and plasma ACE activity. Also no correlation was found between high or low ACE activity level and first dose hypotension. The practical importance of the results are: For patients with congestive heart failure. FOS and CAP have different effects on BP after the first dose, and this effect may be dependent on the plasma ACE activity level. FOS produces ACE inhibition and BP changes similar to placebo so it is the safer choice for the treatment of congestive heart failure. PMID- 11384080 TI - Effects of propofol on ischemia and reperfusion in the isolated rat heart compared with thiamylal. AB - The aim of the present study was to investigate whether clinical doses of propofol and thiamylal affect oxygen free radical production and intracellular calcium concentration ([Ca2+]i) in the post-ischemic reperfused heart. Forty eight rat hearts were perfused with a Langendorff system and loaded with Fura-2 / AM as a [Ca2+]i marker. The hearts were divided into 6 groups as follows (each group: n = 8); Group S (saline), Group TL (thiamylal 100 microM), Group TH (thiamylal 300 microM), Group I (Intralipid), Group PL (propofol 3 microM), and Group PH (propofol 10 microM). All hearts were initially perfused for 5 min as control aerobic perfusion. Afterwards, no-flow ischemia was induced for 15 min, followed by reperfusion for 20 min. The formation of hydroxyl radicals in the coronary effluent was measured with high performance liquid chromatography using salicylic acid. At the beginning of the ischemia and reperfusion periods, increases in systolic and diastolic [Ca2+]i were observed in all groups except Group TH. The high dose of thiamylal significantly suppressed this initial increase in cytosolic [Ca2+]i (Group S 1.30+/-0.15; Group TL 0.99+/-0.17; Group TH 0.70+/-0.09, at 1 min after reperfusion; systolic [Ca2+]i : p < 0.05). Total DHBAs in the coronary effluent of all groups increased significantly 1 min after reperfusion, however, there were no significant differences among the groups. Clinical doses of propofol had no significant effect on myocardial function and [Ca2+]i before and after ischemia, whereas thiamylal suppressed the increase in [Ca2+]i during ischemia and reperfusion. However, free radical formation during reperfusion was unaffected by thiamylal and propofol. PMID- 11384081 TI - Beat dependent alteration of Ca2+-activated Cl- current during rapid stimulation in rabbit ventricular myocytes. AB - The transient outward currents (Ito) play an important role in action potential repolarization in cardiac myocytes. Two components of Ito have been identified as 4-AP-sensitive but Ca2+-insensitive Ito carried by K, and Ca2+-sensitive but 4-AP insensitive Ito carried by Cl- (I(Cl(Ca))). It is known that the amplitudes of Ito change depending on the stimulation frequency. In this study we investigated the beat dependent alteration of I(Cl(Ca)) during rapid stimulation using the whole cell patch clamp technique in rabbit ventricular myocytes. The cells were internally perfused with a solution containing 0.1 microM free Ca2+ to develop I(Cl(Ca)) and all internal K+ was replaced with Cs+ to block 4-AP-sensitive Ito and other K+ currents. By applying depolarizing pulses at a high frequency of 2.5 Hz, the amplitudes of I(Cl(Ca)) gradually increased as the number of pulses increased following a transient decrease in the 2nd pulse and reached a plateau level at the 20th pulse. The shape of the current-voltage curve of I(Cl(Ca)) was not overly different for different numbers of preceding pulses. The recovery from inactivation of I(Cl(Ca)) could be fitted to a single exponential curve and full recovery was achieved after > 1 sec with a time constant of 368 ms. The ramp clamp experiments showed that the conductance of the background I(Cl(Ca)) increased with the preceding pulse numbers, indicating that the resting level of [Ca2]i increased with the pulses applied. From these results, we conclude that beat dependent alteration of I(Cl(Ca)) is determined by not only its apparent kinetic property, but also the resting level of [Ca2+]i during rapid stimulation. PMID- 11384082 TI - Vascular smooth muscle cell proliferation is effectively suppressed by the non specific growth factor inhibitor suramin. AB - The purpose of this study was to investigate the effects of the non-specific growth factor inhibitor suramin on smooth muscle cell proliferation in vitro and in vivo. Cultured vascular smooth muscle cells (VSMC) were stimulated by platelet derived growth factor (PDGF) and cellular DNA synthesis assessed by [3H] thymidine uptake. Suramin dose-dependently inhibited DNA synthesis in VSMC, and 100 microM of suramin completely suppressed the PDGF-AB-induced cellular DNA synthesis. Rabbit carotid arteries were injured by the balloon catheter, and then suramin locally delivered using a porous balloon catheter over ten minutes. Three weeks after the vascular injury, the extent of intimal thickening was compared between the suramin-treated and control rabbits. The neointimal formation triggered by balloon-mediated vascular injury was suppressed significantly and dose-dependently by locally infused suramin, and the intima to media area ratios of the control and 1 mM suramin-treated animals were 48.8+/-14.9 and 12.2+/-6.0%, respectively (p < 0.01. n = 6 for each group). These results suggest that one time local administration of suramin was sufficient to suppress neointimal formation after balloon-mediated vascular injury, and that pharmacological intervention targeting the growth factor's signaling pathways could be a promising approach to prevent smooth muscle cell proliferation in various proliferative vascular diseases. PMID- 11384083 TI - Clinicopathological characteristics of left ventricular myocardium with transient asynergy: report of three cases. AB - The aim of the present retrospective study was to clarify the histopathologic substrates of left ventricular myocardium with transient asynergy due to acute ischemic insult in man. Three patients who had had prolonged chest pain, new abnormal Q waves and new ST segment elevation were studied. There was no significant elevation of serum creatine phosphokinase activity in two of the three patients. Echocardiograms on admission or the next day showed severe hypokinetic or akinetic motion and thinning of the anteroseptal and apical portions of the left ventricle and regional dilatation of the same portions. Disappearance of the abnormal Q waves, ST segment elevation resolution, and early T wave inversion were observed later. Complete improvement of the echocardiographic abnormalities was confirmed after a few weeks in all patients. Manifest ischemic lesions of subendocardial scars of the anteroseptal region of the left ventricle were detected in only one of the three cases by gross examination. However, on microscopic examination, islands of necrotic myocytes were interspersed with islands of viable cells throughout the jeopardized region in one case, although the scattered necrotic foci were restricted to the subendocardium and the trabeculae. Normal myocardium and subendocardial scars were observed in the other two cases. In conclusion, left ventricular myocardium with transient asynergy. detected clinically during acute ischemic attack, consists of normal myocardium or small ischemic lesions primarily in the subendocardium. PMID- 11384084 TI - Effectiveness of an antiplatelet agent for coronary artery ectasia associated with silent myocardial ischemia. AB - A 74-year-old Japanese male was referred to our hospital because of an abnormal electrocardiogram. The electrocardiogram revealed tall P waves in leads II, III, and aV(F). Echocardiography disclosed hypokinesis extending from the anteroseptal region to the apex. Iodine-123 15-(p-iodophenyl)-3-(R,S)-methylpentadecanoic acid (123-BMIPP) scintigraphy revealed reduced uptake from the anteroseptal region to the apex. Coronary arteriography demonstrated diffuse dilatation of the right and left coronary arteries without organic stenosis, and left ventriculography showed hypokinesis at the same area. Furthermore, the coronary flow reserve in the left anterior descending artery was decreased. He was treated with an antiplatelet agent. Ten months later, the left ventriculography, 123I-BMIPP scintigraphy findings and coronary flow reserve were normalized. These findings demonstrate that antiplatelet therapy may be useful in the preservation of left ventricular function in patients with coronary artery ectasia. PMID- 11384085 TI - Hypoglycemic syncope induced by a combination of cibenzoline and angiotensin converting enzyme inhibitor. AB - A 65-year-old Japanese woman with dilated cardiomyopathy, hypothyroidism and refractory sustained ventricular tachycardia experienced a near-death hypoglycemic syncope. The attack seemed to be induced by a high level of serum insulin, probably due to cibenzoline and by concomitant use of an angiotensin converting enzyme inhibitor (ACEI). Additionally, decreased food intake because of a severe toothache may have contributed to the deterioration of her condition. This case warns cardiologists that a combined cibenzoline and ACEI therapy can provoke serious adverse effects such as hypoglycemic syncope in the elderly. Therefore, the possibility of a hypoglycemic attack associated with these drugs should be explained to patients who are in poor condition. PMID- 11384086 TI - Allograft aortic root replacement for aortic valve endocarditis with aortopulmonary fistula. AB - Acute infective endocarditis affecting the aortic root and valve associated with development of a fistulous communication between the aorta and pulmonary artery was presented in a young Turkish girl. Emergency surgery was required. Operation consisted initially of closure of the defect on the main pulmonary artery with a pericardial patch. This was followed by allograft aortic root replacement. PMID- 11384087 TI - Left ventricular apical aneurysm in cardiac sarcoidosis. AB - A 53-year-old woman was hospitalized for general fatigue and palpitations. An electrocardiogram showed ST elevation and T wave inversion in leads II, III, aVF, and V4-6. Cardiac catheterization was performed since the echocardiogram demonstrated the existence of a left ventricular apical aneurysm. Left ventriculography showed an aneurysm of the apex. An endomyocardial biopsy specimen from the left ventricular apical wall demonstrated typical noncaseating granulomas with giant cells. The patient was diagnosed as having cardiac sarcoidosis. There was no evidence suggesting involvement of other systemic organs. Cardiac sarcoidosis should be considered within a spectrum of diseases that cause left ventricular aneurysm. PMID- 11384088 TI - Developing effective cancer vaccines: design and monitoring are critical. PMID- 11384089 TI - Oral fluoropyrimidines among the new drugs for patients with metastatic breast cancer. AB - Although drugs such as the taxoids and vinorelbine have increased the options available for anthracycline-resistant metastatic breast cancer, new therapeutic options are needed, particularly for taxoid-refractory tumours. Increasing emphasis is being placed on the development of oral agents, which many patients prefer provided efficacy is not compromised, particularly if the oral agents are less toxic than current intravenous agents. Capecitabine, a new, oral fluoropyrimidine, mimics continuous infusion 5-FU and is activated preferentially at the tumour site. Phase II studies of capecitabine have demonstrated encouraging response rates in patients with few further treatment options (20% response with an additional 43% achieving stable disease in paclitaxel-refractory patients; 36% response with a further 23% achieving stable disease in anthracycline-refractory patients). In addition, a randomized, phase II trial demonstrated a response rate of 30% (95% Cl: 19-43%) with capecitabine as first line treatment for metastatic breast cancer, compared with 16% (95% Cl: 5-33%) in patients receiving low-dose CMF. These trials also showed that capecitabine has a favourable safety profile typical of infused fluoropyrimidines. Both alopecia and myelosuppression were rare. Capecitabine may therefore provide an effective, well tolerated and convenient alternative to intravenous cytotoxic agents, not only in taxoid-resistant patients, but also in anthracycline-resistant metastatic breast cancer or as first-line therapy. Furthermore, the low incidence of myelosuppression makes capecitabine an attractive agent for incorporation into combination regimens with agents such as epirubicin/doxorubicin, the taxoids and vinorelbine. PMID- 11384091 TI - A national survey of the chemotherapy regimens used to treat small cell lung cancer (SCLC) in the United Kingdom. AB - Many chemotherapy regimens are used for treating SCLC in the United Kingdom, but it is not known, in any detail, which regimens are used, by which specialists, for which types of patient. We conducted a survey among all medical and clinical oncologists, respiratory physicians and general physicians with respiratory interest in the United Kingdom to find out. The questionnaire asked for the number of SCLC patients treated annually; how many were given chemotherapy; the drugs, doses and schedules chosen according to prognostic group (as defined by the clinician); and the reasons for choice of regimen. 1214 questionnaires were sent out, and responses were received from 1070 (88%) clinicians; 266 (25%) of these treated SCLC with chemotherapy. Of 4674 patients given chemotherapy annually, 36% were given it by clinical oncologists, 30% by medical oncologists, 27% by respiratory physicians, and 7% by general physicians. In all, 34 regimens were reported with 151 different combinations of dose and schedule. In 2311 good prognosis patients, 23 regimens were used, the commonest being ACE (doxorubicin, cyclophosphamide, etoposide), ICbE (ifosfamide, carboplatin, etoposide), CAV (cyclophosphamide, doxorubicin, vincristine), CbE (carboplatin, etoposide), and PE (cisplatin, etoposide). In 1517 poor prognosis patients, 21 regimens were used, the commonest being CAV, EV (etoposide, vincristine), CbE, CAV alternating with PE, and oral etoposide. 452 patients were treated regardless of prognosis and for 219 no prognostic criteria were specified. The remaining 175 were given second-line chemotherapy or were given regimens chosen to avoid toxicity or because of intercurrent disease or other reasons. The main reasons affecting choice of regimen were routine local practice, patients' convenience, quality of life considerations, trial results and cost. The results show wide variation in routine practice and will be useful in reporting and planning clinical trials and in deciding on local treatment policies. PMID- 11384090 TI - Randomized double-blind phase II survival study comparing immunization with the anti-idiotypic monoclonal antibody 105AD7 against placebo in advanced colorectal cancer. AB - The cancer vaccine 105AD7 is an anti-idiotypic monoclonal antibody that mimics the tumour-associated antigen 791T/gp72 (CD55, Decay Accelerating Factor) on colorectal cancer cells. Phase I studies in patients with advanced disease confirmed that 105AD7 is non-toxic, and that T cell responses could be generated. A prospective, randomized, double-blind, placebo-controlled survival study in patients with advanced colorectal cancer was performed. 162 patients were enrolled between April 1994 and October 1996. Patients attended at trial entry, and at 6 and 12 weeks, where they received 105AD7 or placebo. Study groups were comparable in terms of patient demographics, and time from diagnosis of advanced colorectal cancer (277.1 v 278.6 days). Baseline disease was similar, with 50% of patients having malignancy in at least 2 anatomic sites. Compliance with treatment was poor, with only 50% of patients receiving 3 planned vaccinations. Median survival from randomization date was 124 and 184 days in 105AD7 and placebo arms respectively (P = 0.38), and 456 and 486 days from the date of diagnosis of advanced disease (P = 0.82). 105AD7 vaccination does not prolong survival in patients with advanced colorectal cancer. The reasons for lack of efficacy are unclear, but may reflect the high tumour burden in the patient population, and poor compliance with immunization. Further vaccine studies should concentrate on patients with minimal residual disease. PMID- 11384092 TI - Feasibility of intraventricular administration of etoposide in patients with metastatic brain tumours. AB - As the systemic administration of etoposide is effective in the treatment of relapsed and metastatic brain tumours, a pilot trial was designed to study the feasibility of intraventricular administration of etoposide in such patients. 14 patients aged 2.1 to 33.2 years were treated with intraventricular etoposide simultaneously with either oral or intravenous chemotherapy with trofosfamide or carboplatin and etoposide. In 59 courses (1-12/patient) 0.5 mg etoposide was administered daily via an indwelling subcutaneous reservoir for 5 consecutive days every 2-5 weeks over a period of 0-11 months. During 15 courses in 5 patients serial CSF samples were obtained and etoposide levels were determined by reversed-phase HPLC. Side effects included transient headache and bacterial meningitis, each during 2 courses. Pharmacokinetic data analysis in the CSF (11 courses, 4 patients) revealed a terminal half-life of 7.4+/-1.2 hours and an AUC of 25.0 +/- 9.5 microg h ml(-1)(mean +/- standard deviation). The volume of distribution at steady state and total clearance exhibited a large interindividual variability with mean values of 0.16 l and 0.46 ml min( 1)respectively. Intraventricularly administered etoposide is well tolerated. CSF peak levels exceed more than 100-fold those achieved with intravenous infusions. Further studies should be focused on optimizing the dose and schedule and on determining the effectiveness of intraventricularly administered etoposide. PMID- 11384093 TI - Childhood cancer incidence in a cohort of twin babies. AB - We studied childhood cancer incidence in a population-based twin cohort using record linkage to the National Registry of Childhood Tumours. After correcting for mortality, an incidence deficit was observed (Standardized Incidence Ratio (SIR) 79; 95% Confidence Interval (CI) 39-120). Pooled analysis with data from published cohort studies indicates a similar significant incidence reduction (SIR 81, 95% CI 67-96). Further studies are warranted. PMID- 11384094 TI - An epidemiological study of cancer in adult twins born in Norway 1905-1945. AB - We have identified 23 334 individuals (40%) of twins born in Norway 1905-45 where both twins were alive in 1960 without malignant disease. These were linked to the Cancer Registry of Norway. A reduced risk of malignant disease was demonstrated among twins for all tumour sites combined; standardized incidence rate (SIR): 0.90 (95% CI 0.85-0.94) in females and 0.95 (95% Cl 0.90-0.99) in males. In both sexes, we observed a significant reduced incidence of malignant melanomas of the skin. The incidence of colorectal cancer tended to be reduced for both sexes. In females, the incidence of tumours of the central nervous system and lungs were reduced. We consider our findings are real, but cannot explain them. PMID- 11384096 TI - Lactation and breast cancer risk: a case-control study in Connecticut. AB - In this report, we examined the relationship between lactation and breast cancer risk, in a case-control study of breast cancer, conducted in Connecticut between 1994 and 1998. Included were 608 incident breast cancer cases and 609 age frequency matched controls, aged 30-80 years old. Cases and controls were interviewed by trained study interviewers, using a standardized, structured questionnaire, to obtain information on lactation and other major risk factors. Parous women who reported ever lactation had a borderline significantly reduced risk of breast cancer (OR = 0.83, 95% CI, 0.63-1.09). An OR of 0.53 (95% CI, 0.27 1.04) was observed in those having breastfed more than 3 children compared to those who never lactated. Women having breastfed their first child for more than 13 months had an OR of 0.47 (95% CI, 0.23-0.94) compared to those who never breastfed. Lifetime duration of lactation also showed a risk reduction while none of the ORs were statistically significant. Further stratification by menopausal status showed a risk reduction related to lactation for both pre- and postmenopausal women, while the relationship is less consistent for the latter. These results support an inverse association between breastfeeding and breast cancer risk. PMID- 11384095 TI - Birth order, family size, and the risk of cancer in young and middle-aged adults. AB - We used the Swedish Family-Cancer Database to analyse the effects of birth order and family size on the risk of common cancers among offspring born over the period 1958-96. Some 1.38 million offspring up to age 55 years with 50.6 million person-years were included. Poisson regression analysis included age at diagnosis, birth cohort, socio-economic status and region of residence as other explanatory variables. The only significant associations were an increasing risk for breast cancer by birth order and a decreasing risk for melanoma by birth order and, particularly, by family size. When details of the women's own reproductive history were included in analysis, birth orders 5-17 showed a relative risk of 1.41. The effects on breast cancer may be mediated through increasing birth weight by birth order. For melanoma, socio-economic factors may be involved, such as limited affordability of sun tourism in large families. Testis cancer showed no significant effect and prostate cancer was excluded from analysis because of the small number of cases. PMID- 11384097 TI - Estimation of screening test (Hemoccult) sensitivity in colorectal cancer mass screening. AB - 3 controlled cohorts of mass-screening for colorectal cancer using a biennial faecal occult blood (HemoccultII test on well-defined European populations have demonstrated a 14% to 18% reduction in specific mortality. We aimed to estimate the sensitivity (S) of this HemoccultII test and and also mean sojourn time (MST) from French colorectal mass-screening programme data. 6 biennial screening rounds were performed from 1988 to 1998 in 45 603 individuals aged 45-74 years in Saone et-Loire (Burgundy, France). The prevalent/incidence ratio was calculated in order to obtain a direct estimate of the product S.MST. The analysis of the proportional incidence and its modelling was used to derive an indirect estimate of S and MST. The product S.MST was higher for males than females and higher for left colon than either the right colon or rectum. The analysis of the proportional incidence confirmed the result for subsites but no other significant differences were found. The sensitivity was estimated at 0.57 and the MST at 2.56 years. This study confirms that the sensitivity of the Hemoccult test is relatively low and that the relatively short sojourn time is in favour of annual screening. PMID- 11384098 TI - Spatial variation and temporal trends of testicular cancer in Great Britain. AB - Increases in testicular cancer incidence have been reported in several countries over a long period. Geographical variability has also been reported in some studies. We have investigated temporal trends and spatial variation of testicular cancer at ages 20-49 in Britain. Temporal trends in testicular cancer incidence were examined, 1974 to 1991 and in mortality, 1981-1997. Spatial variation in incidence was analysed across electoral wards, 1975 to 1991. We used Poisson regression to examine for regional and socio-economic effects and Bayesian mapping techniques to analyse small-area spatial variability. Incidence increased from 6.5 to 11.1 per 100 000 in men at ages 20-34, and from 5.6 to 9.7 per 100 000 in men at ages 35-49, while mortality declined by 50% in both age groups. Risks of testicular cancer varied across regional cancer registries, ranging from 0.79 (95% CI: 0.73-0.84) to 1.32 (95% CI: 1.25-1.38), and was higher in the most affluent compared with the most deprived areas. Analyses within 2 regions (one predominantly urban, the other predominantly rural) did not indicate any localized geographical clustering. The increasing incidence contrasted with a decreasing mortality over time in Great Britain, similar to that found in other countries. The higher risk in more affluent areas is not consistent with findings on social class at the individual level. The absence of any marked geographical variability at small area scale argues against a geographically varying environmental factor operating strongly in the aetiology of testicular cancer. PMID- 11384099 TI - Overexpression of matrix-metalloproteinase-9 in human breast cancer: a potential favourable indicator in node-negative patients. AB - Matrix metalloprotease-9 (MMP-9; 92 kDa type IV collaganase, gelatinase B) is regarded as, important for degradation of the basement membrane and extracellular matrix during cancer invasion and other tissue-remodelling events. In this study we evaluate the prognostic value of MMP-9, by immunoperoxidase staining in a series of 210 breast cancer tissues. The results were quantitated using the HSCORE system, which consider both staining intensity and the percentage of cells stained at given intensities. MMP-9 status was compared with the concentration of cytosolic Cathepsin-D and with other established prognostic factors, in terms of disease free survival and overall survival. The median follow-up period was 62 months. MMP-9 staining was observed primarily in cancer cells, and to a lesser degree in surrounding stromal cells. MMP-9 expression was not detected in normal breast tissue. Levels of MMP-9 expression below the cut-off point were more frequently observed in larger (P = 0.014), invasive ductal histologic (P = 0.037), progesterone receptor (PR)-negative and PR-strong positive tumours (P< 0.001), as well as samples belonging to patients with stage III-IV disease (P = 0.009) and age 45-55 years (P = 0.011). In univariate analysis, node-negative breast cancer patients with tumors positive for MMP-9 had a considerable reduction in risk for relapse (RR = 0.45;P = 0.039) or death (RR = 0.32;P = 0.009). Multivariate analysis indicated that MMP-9 status was an independent favourable predictor of OS (RR = 0.47;P = 0.034) in node-negative but not in node positive patients. Our results suggest that MMP-9 may be an independent favourable prognostic factor in node-negative breast cancer patients. The overexpression of MMP-9 in breast cancer may be also used as a marker to subdivide node negative breast cancer patients in order to determine the optimal treatment modality. PMID- 11384100 TI - Frequent p53 mutation in brain (fetal)-type glycogen phosphorylase positive foci adjacent to human 'de novo' colorectal carcinomas. AB - 'de novo' carcinogenesis has been advocated besides 'adenoma carcinoma sequence' as another dominant pathway leading to colorectal carcinoma. Our recent study has demonstrated that the distribution of brain (fetal)-type glycogen phosphorylase (BGP) positive foci (BGP foci) has a close relationship with the location of 'de novo' carcinoma. The aims of the present study are to investigate genetic alteration in the BGP foci and to characterize them in the 'de novo' carcinogenesis. 17 colorectal carcinomas without any adenoma component expressing both immunoreactive p53 and BGP protein were selected from 96 resected specimens from our previous study. Further investigations to examine the proliferating cell nuclear antigen (PCNA)-labelling index, and the p53 and the codon 12 of K-ras mutation using the polymerase chain reaction-single strand conformation polymorphism were performed in the BGP foci, BGP negative mucosa and carcinoma. The BGP foci were observed sporadically in the transitional mucosa adjacent to the carcinoma in all cases. The PCNA labelling index in the BGP foci was significantly higher than that in the BGP negative mucosa (P< 0.001). p53 mutations were observed in 8 carcinomas, but no K-ras mutation was detected. Interestingly, although none of the overexpressions of p53 protein was detected immunohistochemically in the BGP positive foci, the p53 gene frequently (41.2% of the BGP foci tested) mutated in spite of no K-ras mutation. The present study demonstrates potentially premalignant foci in the colorectal transitional mucosa with frequent p53 gene mutation. It is suggested that BGP foci are promising candidates for the further investigation of 'de novo' colorectal carcinogenesis. PMID- 11384101 TI - p53 mutations in urinary bladder cancer. AB - We have screened for mutations in exons 5-8 of the p53 gene in a series consisting of 189 patients with urinary bladder neoplasms. 82 (44%) neoplasms were lowly malignant (Ta, G1-G2a) and 106 (56%) were highly malignant (G2b-G4 or > or = T1). Only one mutation was in a lowly malignant urinary bladder neoplasm, in total we found p53 mutations in 26 (14%) of the 189 patients. 30% of the samples had loss of heterozygosity (LOH) for one or both of the p53 exogenic (CA)n repeat and the p53 intragenic (AAAAT)n repeat markers. 31 samples (21%) showed LOH but were not mutated, suggesting other mechanisms inactivating p53 than mutations. 4 mutations were found at codon 280 and 2 mutations were found at codon 285, 2 previously reported hot spots for urinary bladder cancer. The study indicate a boundary between G2a and G2b tumours concerning the occurrence of genetic events affecting p53 function; moderately differentiated (G2) urinary bladder neoplasms probably are genetically heterogeneous which supports the suggestion that they should not be grouped together but instead, for example, be categorized as either lowly or highly malignant. PMID- 11384102 TI - Identification of potential diagnostic markers of prostate cancer and prostatic intraepithelial neoplasia using cDNA microarray. AB - The identification of novel genes or groups of genes expressed in prostate cancer may allow earlier diagnosis or more accurate staging of the disease. We describe the assembly and use of a 1877-member microarray representing cDNA clones from a range of prostate cancer stages and grades, precursor lesions and normal tissue. Using labelled cDNA from tumour samples obtained from TURP or radical prostatectomy, analysis of expression patterns identified many up-regulated transcripts. Cell lines were found to over-express fewer genes than diseased tissue samples. 17 known genes were found to over-express more than 4-fold in 4 or more cancers out of 15 cancers. Only 2 genes were over-expressed in 6 out of 15 cancers or more, whilst no genes were consistently found to be over-expressed in all cancer samples. Novel prostate cancer associations for several well characterized genes or full length cDNAs were identified, including PLRP1, JM27, human UbcM2, dynein light intermediate chain 2 and human homologue of rat sec61. Novel associations with high-grade PIN include: breast carcinoma fatty acid synthase and cDNA DKFZp434B0335. We shortlist and discuss the most significant over-expressed genes in prostate cancer and PIN, and highlight expression differences between malignant and benign samples. PMID- 11384103 TI - Identification of a 170-kDa protein over-expressed in lung cancers. AB - Lung cancer is the leading cause for cancer death in both male and female populations. Although many molecular markers for lung cancer have been developed and useful for early detection of lung cancer, their function remains unknown. In this paper, we report our findings that a 170-kDa protein (p170) is over expressed in all types of human lung cancers compared with normal tissues and it is identified as a subunit of translation initiation factor eIF3 by cDNA cloning. Translation initiation factors are a family of proteins that promote the initiation step of protein synthesis and are regulators of cell growth at the translational level. Further studies showed that p170 mRNA is ubiquitously expressed with higher levels in adult proliferating tissues (e.g. bone marrow) and tissues during development (e.g. fetal tissues). This study suggests that p170 and eIF3 may be important factors for cell growth, development, and tumorigenesis. PMID- 11384104 TI - Decrease in drug accumulation and in tumour aggressiveness marker expression in a fenretinide-induced resistant ovarian tumour cell line. AB - We investigated whether the efficacy of fenretinide (HPR) against ovarian tumours may be limited by induction of resistance. The human ovarian carcinoma cell line A2780, which is sensitive to a pharmacologically achievable HPR concentration (IC(50)= 1 microM), became 10-fold more resistant after exposure to increasing HPR concentrations. The cells (A2780/HPR) did not show cross-resistance to the synthetic retinoid 6-[3-adamantyl-4-hydroxyphenyl]-2-naphthalene carboxylic acid (CD437) and were not sensitive, similarly to the parent line, to all-trans retinoic acid, 13-cis-retinoic acid or N-(4-methoxyphenyl)retinamide. A2780/HPR cells showed, compared to parental cells, a 3-fold reduction in colony-forming ability in agar. The development of HPR resistance was associated with a marked increase in retinoic acid receptor beta (RARbeta) mRNA and protein levels, which decreased, together with drug resistance, after drug removal. The expression of cell surface molecules associated with tumour progression including HER-2, laminin receptor and beta1 integrin was markedly reduced. The increase in the levels of reactive oxygen species is not involved in HPR-resistance because it was similar in parental and resistant cells. Conversely differences in pharmacokinetics may account for resistance because, in A2780/HPR cells, intracellular peak drug levels were 2 times lower than in A2780 cells and an as yet unidentified polar metabolite was present. These data suggest that acquired resistance to HPR is associated with changes in marker expression, suggestive of a more differentiated status and may be explained, at least in part, by reduced drug accumulation and increased metabolism. PMID- 11384106 TI - Cytotoxicity of ascorbate, lipoic acid, and other antioxidants in hollow fibre in vitro tumours. AB - Vitamin C (ascorbate) is toxic to tumour cells, and has been suggested as an adjuvant cancer treatment. Our goal was to determine if ascorbate, in combination with other antioxidants, could kill cells in the SW620 hollow fibre in vitro solid tumour model at clinically achievable concentrations. Ascorbate anti-cancer efficacy, alone or in combination with lipoic acid, vitamin K3, phenyl ascorbate, or doxorubicin, was assessed using annexin V staining and standard survival assays. 2-day treatments with 10 mM ascorbate increased the percentage of apoptotic cells in SW620 hollow fibre tumours. Lipoic acid synergistically enhanced ascorbate cytotoxicity, reducing the 2-day LC(50)in hollow fibre tumours from 34 mM to 4 mM. Lipoic acid, unlike ascorbate, was equally effective against proliferating and non-proliferating cells. Ascorbate levels in human blood plasma were measured during and after intravenous ascorbate infusions. Infusions of 60 g produced peak plasma concentrations exceeding 20 mM with an area under the curve (24 h) of 76 mM h. Thus, tumoricidal concentrations may be achievable in vivo. Ascorbate efficacy was enhanced in an additive fashion by phenyl ascorbate or vitamin K3. The effect of ascorbate on doxorubicin efficacy was concentration dependent; low doses were protective while high doses increased cell killing. PMID- 11384105 TI - Inhibition of protein farnesylation enhances the chemotherapeutic efficacy of the novel geranylgeranyltransferase inhibitor BAL9611 in human colon cancer cells. AB - Proteins belonging to the ras superfamily are involved in cell proliferation of normal and neoplastic tissues. To be biologically active, they require post translational isoprenylation by farnesyl-transferase and geranylgeranyl transferase. Enzyme inhibition by drugs may thus represent a promising approach to the treatment of cancer. Therefore, the combined effect of BAL9611, a novel inhibitor of geranylgeranylation, and manumycin, a farnesyl-transferase inhibitor, was evaluated on the SW620 human colon cancer cell line which harbours a mutated K-ras gene. BAL9611 and manumycin dose-dependently inhibited SW620 cell growth with 50% inhibitory concentration (IC(50)) of 0.47 +/- 0.03 and 5.24 +/- 1.41 microM (mean +/- SE), respectively. The isobologram analysis performed at the IC(50)level revealed that the combined treatment was highly synergistic with respect to cell growth inhibition. BAL9611 and manumycin were able to inhibit the geranylgeranylation of p21rhoA and farnesylation of p21ras; both drugs inhibited p42ERK2/MAPK phosphorylation, but their combination was more effective than either drug alone. Moreover, the enhanced inhibition of cell growth in vitro by the BAL9611-manumycin combination was also observed in vivo in CD nu/nu female mice xenografted with SW620 tumours. Finally, both drugs were able to induce cell death by apoptosis in vitro and in vivo, as demonstrated by perinuclear chromatin condensation, cytoplasm budding and nuclear fragmentation, and interoligonucleosomal DNA digestion. In conclusion, the inhibition of protein farnesylation enhances the chemotherapeutic effect of BAL9611 in vitro and in vivo in a synergistic fashion, as a result of the impairment of post translational isoprenylation of proteins and phosphorylation of p42ERK2/MAPK, whose activation is associated with post-translational geranylgeranylation and farnesylation of p21rhoA and p21ras. PMID- 11384107 TI - Telomerase activity and p53-dependent apoptosis in ovarian cancer cells. AB - We conducted the present study to determine the relationship between p53 dependent apoptosis and telomerase activity in ovarian cancer cells. A human ovarian adenocarcinoma cell line, SK-OV-3 that had homozygous deletion of the p53 gene was used in this study. Wild-type p53 genes were transducted to SK-OV-3 cells with a recombinant adenovirus that contained a wild-type p53 gene (AxCAp53). IC(50)to cisplatin (CDDP) was 12.9 microM for SK-OV-3 cells and 9.2 microM for p53 gene-transducted SK-OV-3 cells. The apoptotic index for cells with p53 gene transduction was significantly higher than cells without transduction. Additionally, p53 gene transduction significantly enhanced CDDP-induced apoptosis. Bax protein in SK-OV-3 cells did not differ before and after exposure to CDDP. In SK-OV-3 cells with transduction of the p53 gene, the expression of p53 and Bax proteins increased after exposure to CDDP. Expression of Bcl-xL decreased after exposure to CDDP in SK-OV-3 cells with and without transduction. The telomerase activity in SK-OV-3 cells with the p53 gene was significantly lower compared with the cells without the p53 gene. CDDP exposure did not affect telomerase activity and human telomerase reverse transcriptase (hTERT) expression in both cell lines. We suggest that the p53 gene may relate to telomerase activity, but that p53-dependent apoptosis does not affect the activity. PMID- 11384108 TI - Regulation of differentiation- and proliferation-inducers on Lewis antigens, alpha-fucosyltransferase and metastatic potential in hepatocarcinoma cells. AB - The expressions of Lewis (Le) antigens, alpha-1,3/1,4 fucosyltransferases (alpha 1,3/1,4 FuTs), and metastatic potential after the treatment of 2 differentiation inducers, all- trans retinoic acid (ATRA), 8-bromo-cyclic 3',5'adenosine monophosphate (8-Br-cAMP); and 2 proliferation inducers, epidermal growth factor (EGF) and phobol-12-myristate-13-acetate (PMA), on 7721 human hepatocarcinoma cell line were studied. Cell adhesion to human umbilical vein endothelial cells (HUVEC), cell migration through transwell and invasion through matrigel were selected as the indexes of metastatic potential-related phenotypes. Using fluorescence-labelled antibodies and flow-cytometric analysis, it was found that 7721 cells mainly expressed sialyl Lewis X (SLe(x)) and a less amount of sialyl dimeric Lewis X (SDLe(x)) antigens on the cell surface. Their expressions were down-regulated by ATRA, and up-regulated by EGF. SLe(x)antigen was also decreased and increased by the treatment of 8-Br-cAMP and PMA respectively. With Northern blot to detect the mRNAs of alpha-1,3/1,4 FuTs, the main enzymatic basis for the change in SLe(x)expression was found to be the alteration of the expression of alpha-1,3 FuT-VII. It was evidenced by the observations that alpha-1,3 FuT-VII was the main alpha-1,3/1,4 FuT in 7721 cells, while alpha-1,3/1,4 FuT-III and alpha-1,3 FuT-VI were expressed rather low. The changes in the expressions of SLe(x)antigen and alpha-1,3 FuT-VII resulted in the altered cell adhesion to tumour necrosis factor-alpha stimulated HUVEC, since only the monoclonal antibody of the SLe(x), but not other monoclonal antibodies blocked the adhesion of 7721 cells to HUVEC. The migration and invasion of 7721 cells were also reduced by the treatment of ATRA or 8-Br-cAMP, and elevated by EGF or PMA. The above findings indicate that the metastatic potential of 7721 cells is suppressed by differentiation-inducers and promoted by proliferation-inducers. PMID- 11384109 TI - Administration route-dependent vaccine efficiency of murine dendritic cells pulsed with antigens. AB - Dendritic cells (DCs) loaded with tumour antigens have been successfully used to induce protective tumour immunity in murine models and human trials. However, it is still unclear which DC administration route elicits a superior therapeutic effect. Herein, we investigated the vaccine efficiency of DC2.4 cells, a murine dendritic cell line, pulsed with ovalbumin (OVA) in the murine E.G7-OVA tumour model after immunization via various routes. After a single vaccination using 1 x 10(6)OVA-pulsed DC2.4 cells, tumour was completely rejected in the intradermally (i.d.; three of four mice), subcutaneously (s.c.; three of four mice), and intraperitoneally (i.p.; one of four mice) immunized groups. Double vaccinations enhanced the anti-tumour effect in all groups except the intravenous (i.v.) group, which failed to achieve complete rejection. The anti-tumour efficacy of each immunization route was correlated with the OVA-specific cytotoxic T lymphocyte (CTL) activity evaluated on day 7 post-vaccination. Furthermore, the accumulation of DC2.4 cells in the regional lymph nodes was detected only in the i.d.-and s.c.-injected groups. These results demonstrate that the administration route of antigen-loaded DCs affects the migration of DCs to lymphoid tissues and the magnitude of antigen-specific CTL response. Furthermore, the immunization route affects vaccine efficiency. PMID- 11384110 TI - Modulation of docetaxel-induced apoptosis and cell cycle arrest by all- trans retinoic acid in prostate cancer cells. AB - We report that all- trans retinoic acid (ATRA) enhanced the toxicity of docetaxel against DU145 and LNCaP prostate cancer cells, and that the nature of the interaction between ATRA and docetaxel was highly synergistic. Docetaxel-induced apoptotic cell death was associated with phosphorylation and hence inactivation of Bcl-2. ATRA enhanced docetaxel-induced apoptosis and combined treatment with ATRA and docetaxel resulted in down-regulation of Bcl-2. Docetaxel caused phosphorylation and hence inactivation of cdc2 kinase result ing in G2/M arrest. ATRA inhibited docetaxel-induced phosphorylation of cdc2 resulting in activation of cdc2 kinase and partial reversal of the G2/M arrest. ATRA also inhibited docetaxel-induced activation of MAPK indicating that the effects of docetaxel and ATRA on cdc2 phosphorylation are dependent on MAPK. We conclude that ATRA synergistically enhances docetaxel toxicity by down-regulating Bcl-2 expression and partially reverses the docetaxel-induced G2/M arrest by inhibiting docetaxel induced cdc2 phosphorylation in a pathway that is dependent on MAPK. PMID- 11384111 TI - Positional changes and stability of bone segments during simultaneous bilateral mandibular lengthening and widening by distraction. AB - The purpose of this study was to analyse the skeletal changes and stability of the distracted segments during and after simultaneous widening and bilateral lengthening of the mandible in baboons with a miniaturized intraoral bone-borne osteodistractor. Distraction appliances were activated 5 days after vertical posterior body and midsymphyseal osteotomies at a rate of 0.9 mm/day for 10 days. The appliances were then stabilized for a period of 8 weeks, after which the animals were killed. The distraction gaps and gingival tissues were studied clinically and on standardized radiographs. The proportional movement of the distracted segments that we found supports the clinical use of the miniaturized intraoral bone-borne distraction appliance to widen and lengthen the mandible selectively. It also supports the concept of positioning the mandibular osteodistractors parallel to the common vector of distraction, which should parallel the corrected maxillary occlusal plane. PMID- 11384112 TI - Reflections on: distraction osteogenesis. Distraction osteogenesis--the future? PMID- 11384113 TI - Reflections on: distraction osteogenesis. Distraction osteogenesis. PMID- 11384114 TI - Audit of a multidisciplinary approach to the care of children with unilateral and bilateral cleft lip and palate. AB - Cleft lip and palate affects the child in many ways, particularly appearance, dental arch relationships, growth of the face, and speech development. The key to successful care is management in a multidisciplinary team adhering to a well designed protocol, and careful audit of results. We present the intermediate outcome audit of 15 patients with complete bilateral and unilateral cleft lip and palate whose condition was managed in a multidisciplinary team according to a strict protocol. We give the results observations of operations of a single surgeon's functional primary surgery over a 6-year period in terms of dental arch relationships, cephalometric analyses, aesthetic assessments, and speech analysis. The results show good early facial growth, with dental arch relationships appropriate for the age and group; we found only minor speech discrepancies, with no patients requiring pharyngoplasty. The results show the importance of multidisciplinary management, the value of keeping to sound surgical protocols, and functional techniques in cleft lip and palate surgery. Our study includes the neglected group of children who have bilateral cleft lip and palate, and it conforms to the style of pan-European projects. PMID- 11384115 TI - Costochondral grafts in reconstruction of the temporomandibular joint after condylectomy: an experimental study in sheep. AB - The purpose of this study was to investigate the effect of costochondral grafts in the temporomandibular joint (TMJ) in sheep. Five pure-bred adult Merino sheep were used. The condyle alone was resected and replaced with a costochondral graft from the 13th rib. The sheep were killed 3 months after operation. The range of jaw movements before and after operation and at death were recorded. The joints were examined radiologically, macroscopically, and histologically. A new condylar head with normal configuration and function developed. Histologically, the chondrocytes were arranged in a fashion similar to that of a normal joint. All inferior joint spaces showed fibrous adhesions between the condylar head and disc. This study showed that, when such grafts are used to replace the condyle in an otherwise normal sheep TMJ, they fused to the ramus and reconstituted a nearly normal, fully functional joint. PMID- 11384116 TI - Effect of partial immobilization on reconstruction of ankylosis of the temporomandibular joint with an autogenous costochondral graft:an experimental study in sheep. AB - The purpose of this study was to show the effect of partial immobilization of a costochondral graft reconstruction of an ankylosed temporomandibular joint (TMJ) in five adult sheep. Ankylosis was induced in all right TMJs. At three months, a graft was inserted and partially immobilized. Three months after the second operation, four sheep were killed by an overdose with pentothal. One sheep was killed at one month because of infection. Functionally, the body weight, which decreases with ankylosis, did not recover after insertion of the graft and the range of jaw movements got worse. Histologically, the grafts were well attached to the mandibular rami in three of four joints. In one joint, the graft showed signs of resorption and a foreign body reaction. We conclude that, if the reconstructed joint is partially immobilized, then there will be a degree of reankylosis. There was also a high failure rate. PMID- 11384117 TI - Death certification in patients whose primary treatment for oral and oropharyngeal carcinoma was operation: 1992-1997. AB - The aim of this study was to report the cause and place of death of patients with oral cancer as recorded by death certification and their survival with regard to comorbidity and age. From the departmental head and neck oncology database, 322 patients were identified with previously untreated oral and oropharyngeal squamous cell carcinoma diagnosed between 1992 and 1997 inclusive. Three-hundred were matched with the Office for National Statistics (ONS) and copies of death certificates generated on 6 March 1999. Of the 286 patients primarily treated by operation, 203 were alive and 83 had died. In 56 (68%), oral cancer was a contributory factor to the patient's death. Patients with a medical history recorded on their admission for operation had similar survival curves compared to those having no problems recorded. Although most patients (n= 46 55%) died in hospital, only 13 (16%) died in the Regional Maxillofacial Unit. Almost half of those patients who died were not recorded as dead on the departmental oncology database. This study shows that it is useful to link with the ONS to obtain accurate data on date and place of death. The death certificate also gives a useful indication of the cause of death and this seems unrelated to preoperative comorbidity. PMID- 11384118 TI - Changing incidence of oral and maxillofacial tumours in East Java, Indonesia 1987 1992. Part 1: Benign tumors. PMID- 11384119 TI - Plunging ranula as a complication of intraoral removal of a submandibular sialolith. AB - Mucous cysts in the submandibular region--so-called 'plunging' ranula--are relatively uncommon. We report a case of a plunging ranula that complicated excision of an intraductal sialolith of the submandibular gland. PMID- 11384120 TI - Treating oral ranula: another case against blanket removal of the sublingual gland. AB - There are a number of occasions when ranula-like lesions of the oral floor can develop that do not originate from the sublingual gland, or that arise from the gland with no tendency towards recurrence. The author advises that the unconditional removal of the sublingual gland should not be the standard treatment for all ranulas, and present four case reports to illustrate these and describe how they should be treated. All the patients were treated successfully with conservative management and retained normal functioning sublingual glands. Except for the management of plunging ranulas, caution and a close examination of the origin of the lesion are prudent before considering excision of sublingual glands for all ranula-like lesions. PMID- 11384121 TI - Surgical treatment and follow-up of solitary bone cyst of the mandible:a report of seven cases. PMID- 11384122 TI - Re: Maier et al. Review of severe osteoradionecrosis treated by surgery alone or surgery with postoperative hyperbaric oxygen. Br J Oral Maxillofac Surg 2000; 38: 167-246. PMID- 11384123 TI - Re: Maier et al. Review of severe osteoradionecrosis treated by surgery alone or surgery with postoperative hyperbaric oxygen. Br J Oral Maxillofac Surg 2000; 38: 167-246. PMID- 11384124 TI - Re: Maier et al. Review of severe osteoradionecrosis treated by surgery alone or surgery with postoperative hyperbaric oxygen. Br J Oral Maxillofac Surg 2000; 38: 167-246. PMID- 11384125 TI - Re: Maier et al. Review of severe osteoradionecrosis treated by surgery alone or surgery with postoperative hyperbaric oxygen. Br J Oral Maxillofac Surg 2000; 38: 167-246. PMID- 11384126 TI - Re: Hodder et al. SPECT bone scintigraphy in the diagnosis and management of mandibular condylar hyperplasia. Br J Oral Maxillofac Surg 2000; 38: 87-93. PMID- 11384127 TI - Audit on impacted wisdom teeth. PMID- 11384128 TI - Re: Webster K, Wilde J. Management of anticoagulation in patients with prosthetic heart valves undergoing oral and maxillofacial operations. Br J Oral Maxillofac Surg 2000; 38: 124-126. PMID- 11384129 TI - Measuring the effects of image interpretation: an evaluative framework. AB - The relaxing of restrictions on reporting films has resulted in radiographers and other health care professionals becoming increasingly involved in the interpretation of images in areas such as mammography, ultrasound and plain film radiography. However, errors and variation in the interpretation of images now represents the weakest area of clinical imaging. This has been highlighted by the difficulty of establishing standards to measure the film reading performance of radiographers as part of role extension initiatives. Despite a growing literature of studies that evaluate the film reading performance of different health care professionals, there is a paucity of evidence of the subsquent effects on the referring clinician's diagnosis, management plans and patient outcome. This paper proposes an evaluative framework that can be used to measure the chain of events from the initial technical assessment of observers' potential to interpret images using search behaviour techniques, through to the potential costs and benefits to society. Evaluating the wider implications of alternative or complementary reporting policies is essential for generating the evidence base to comprehensively underpin policy and practice and direct future research. Brealey, S.(2001). Clinical Radiology56, 341-347. PMID- 11384130 TI - Measuring the effects of image interpretation: an evaluative framework. PMID- 11384132 TI - The utility of computed tomography in acute small bowel obstruction. AB - Small bowel obstruction is a significant cause of acute surgical admissions. Surgeons are tending to favour an initial trial of conservative management. Due to the unreliability of clinical signs to predict accurately those patients requiring early intervention there is an increasing tendency to utilize imaging investigations, particularly computed tomography (CT), to help define the severity, cause and complications of acute small bowel obstruction. The aim of this pictorial review is to demonstrate the contribution CT can make to the management of these patients. Burkill, G. J. C., Bell, J. R. G. & Healy, J. C. (2001). Clinical Radiology56, 350-359. PMID- 11384133 TI - Magnetic resonance imaging of scoliosis. AB - The introduction of magnetic resonance imaging (MRI) has greatly improved the evaluation of patients with scoliosis allowing assessment of any underlying intraspinal abnormalities. The aim of this pictorial review is to illustrate the magnetic resonance appearances of various abnormalities encountered when imaging developmental spinal deformity. Redla, S.et al. (2001). Clinical Radiology56, 360 371. PMID- 11384134 TI - Isolated scaphotrapeziotrapezoid osteoarthritis: prevalence, symptomatology and associated scapholunate ligament disruption in a population presenting to an accident and emergency department with acute wrist injuries. AB - AIM: To determine the prevalence of isolated scaphotrapeziotrapezoid osteoarthritis in a population presenting to an Accident and Emergency Department of Leicester Royal Infirmary with acute wrist injuries. Also to identify the presence of scapholunate ligament disruption in this patient group and quantify symptoms and loss of function in terms of the modified system of Green and O'Brien, a recognized clinical scoring system. MATERIALS AND METHODS: A total of 1711 radiographs of patients attending the Accident and Emergency Department were prospectively reviewed over a 5-month period. Those patients with isolated scaphotrapeziotrapezoid osteoarthritis were invited for clinical review. RESULTS: Sixteen patients were identified with isolated scaphotrapeziotrapezoid osteoarthritis. Two had a poor Green and O'Brien score and evidence of scapholunate ligament disruption (P < 0.05). CONCLUSION: Isolated scaphotrapeziotrapezoid osteoarthritis has a prevalence of 1% in a population presenting to an Accident and Emergency Department with acute wrist injuries over the age of 30 years. Isolated scaphotrapeziotrapezoid osteoarthritis may be asymptomatic even though the changes in the joint are severe. Scapholunate ligament disruption is associated with a poor Green and O'Brien score, but is not present in the majority of cases. Higginson, A. P.et al. (2001). Clinical Radiology56, 372-374. PMID- 11384135 TI - Contributions of an adiabatic initial inversion pulse and K-space re-ordered by inversion-time at each slice position (KRISP) to control of CSF artifacts and visualization of the brain in FLAIR magnetic resonance imaging. AB - AIM: The aim of this study was to compare the performance of three fluid attenuated inversion recovery (FLAIR) pulse sequences for control of cerebrospinal fluid (CSF) and blood flow artifacts in imaging of the brain. The first of these sequences had an initial sinc inversion pulse which was followed by conventional k-space mapping. The second had an initial sinc inversion pulse followed by k-space re-ordered by inversion time at each slice position (KRISP) and the third had an adiabatic initial inversion pulse followed by KRISP. MATERIALS AND METHODS: Ten patients with established disease were studied with all three pulse sequences. Seven were also studied with the adiabatic KRISP sequence after contrast enhancement. Their images were evaluated for patient motion artifact, CSF and blood flow artifact as well as conspicuity of the cortex, meninges, ventricular system, brainstem and cerebellum. The conspicuity of lesions and the degree of enhancement were also evaluated. RESULTS: Both the sinc and adiabatic KRISP FLAIR sequences showed better control of CSF and blood flow artifacts than the conventional FLAIR sequence. In addition the adiabatic KRISP FLAIR sequence showed better control of CSF artifact at the inferior aspect of the posterior fossa. The lesion conspicuity was similar for each of the FLAIR sequences as was the degree of contrast enhancement to that shown with a T(1)weighted spin echo sequence. CONCLUSION: The KRISP FLAIR sequence controls high signal artifacts from CSF flow and blood flow and the adiabatic pulse controls high signal artifacts due to inadequate inversion of the CSF magnetization at the periphery of the head transmitter coil. The KRISP FLAIR sequence also improves cortical and meningeal definition as a result of an edge enhancement effect. The effects are synergistic and can be usefully combined in a single pulse sequence. Curati, W. L.et al. (2001)Clinical Radiology56, 375-384 PMID- 11384136 TI - False-negative breast screening assessment: what lessons can we learn? AB - AIM: To review women who have had breast cancer diagnosed following previous assessment of a screen-detected mammographic abnormality in order to ascertain the frequency and characteristics of false-negative assessment. MATERIALS AND METHODS: The assessment process was reviewed in the study population of 28 women. This included the nature of the lesion recalled for assessment, additional mammography, clinical and ultrasound findings, and the results of fine needle aspiration cytology and needle histology. RESULTS: The frequency of false negative assessment was approximately 0.56%. The median time between false negative assessment and diagnosis of breast cancer was 33 months. The most common mammographic lesion resulting in false-negative assessment was micro calcification seen in 12 cases (43%). Only five of these 12 cases had image guided biopsy, the remainder were thought to be benign on magnification views. Other mammographic abnormalities were nine masses (32%), five architectural distortions (18%) and two asymmetric densities (7%). Of the 16 women with mammographic lesions other than micro-calcifications 10 had a normal ultrasound. CONCLUSION: Radiological interpretation of indeterminate micro-calcifications as benign or malignant is unreliable. An isolated cluster of micro-calcification requires image-guided core biopsy with representative micro-calcification obtained on specimen radiography. Further mammography done at assessment, particularly paddle compression views, should be carefully analysed to ensure areas of architectural distortion have truly resolved. If one imaging modality shows a significant abnormality and another does not the cases must be managed on the basis of the abnormal finding. Burrell, H.C.et al. (2001). Clinical Radiology56, 385-388. PMID- 11384137 TI - Herniography: a prospective, randomized study between midline and left iliac fossa puncture techniques. AB - AIM: To determine whether an optimal site of injection exists for herniography. MATERIALS AND METHODS: This was a prospective, randomized study of 93 consecutive patients who were referred for herniography over a period of 9 months. Patients underwent either a left iliac fossa (LIF) or midline puncture. Parameters assessed included initial adequate needle placement, complications, pain scores and body mass index (BMI). The groups were compared using Chi-squared test for categorical data, Student's t-test for continuous data and the Mann-WhitneyU-test for skewed data, withP < 0.05 considered statistically significant RESULTS: Four complications were encountered (4%), and these were equally distributed between the two groups. Adequate initial positioning of the needle was similar in both groups. The volume of local anaesthetic used was correlated with discomfort using a pain scale: a volume of >6 ml resulted in significantly more pain. More frequent initial adequate needle placement was observed in thin patients (BMI < 45 kg/m(2)) with experienced operators. Conversely, increased body mass index resulted in more difficult needle placement. CONCLUSION: Herniography is a safe procedure with few complications. There was no significant difference comparing the midline and LIF approaches.Nadkarni, S.et al. (2001). Clinical Radiology56, 389-392. PMID- 11384138 TI - Blood pressure changes during barium enema. AB - AIMS: To document blood pressure changes during barium enema examination and to determine at what point in the examination changes are likely to occur. METHODS AND RESULTS: Blood pressure measurements were taken at seven points during the course of barium enema examination in 107 consecutive patients. We found that patients over the age of 60 years had statistically significant decreases in blood pressure when they were stood up during the course of the examination. Many of these patients were asymptomatic. Patients who had symptoms (15/107, 14%) when standing up had a degree of hypotension. The duration of barium enema examination is longer in those patients who experience symptoms. CONCLUSION: During a barium enema examination hypotension occurs at the point of standing up more frequently in patients over 60 years and in those who suffer symptoms at this time. Patients who fall into one of these groups should be considered at risk of fainting at this point in the examination. A modified technique to avoid standing should be considered in at-risk patients. Roach, S. C.et al. (2001). Clinical Radiology56, 393-396. PMID- 11384139 TI - Do preliminary chest X-ray findings define the optimum role of pulmonary scintigraphy in suspected pulmonary embolism? AB - AIM: To investigate if preliminary chest radiograph (CXR) findings can define the optimum role of lung scintigraphy in subjects investigated for pulmonary embolism (PE). MATERIALS AND METHODS: The CXR and scintigraphy findings from 613 consecutive subjects investigated for suspected PE were retrieved from a radiological database. Of 393 patients with abnormal CXRs, a subgroup of 238 was examined and individual radiographic abnormalities were characterized. CXR findings were related to the scintigraphy result. RESULTS: Scintigraphy was normal in 286 subjects (47%), non-diagnostic in 207 (34%) and high probability for PE in 120 (20%). In 393 subjects (64%) the preliminary CXR was abnormal and 188 (48%) of scintigrams in this group were non-diagnostic. Individual radiographic abnormalities were not associated with significantly different scintigraphic outcomes. If the preliminary CXR was normal (36%), the proportion of non-diagnostic scintigrams decreased to 9% (19 of 220 subjects) (P < 0.05). CONCLUSION: In subjects investigated for PE, an abnormal CXR increases the prevalence of non-diagnostic scintigrams. A normal pre-test CXR is more often associated with a definitive (normal or high probability) scintigram result. The chest radiograph may be useful in deciding the optimum sequence of investigations.Forbes, K. P. N., Reid, J. H., Murchison, J. T.(2001). Clinical Radiology56, 397-400. PMID- 11384140 TI - Barium enema and endoscopy for the detection of colorectal neoplasia: sensitivity, specificity, complications and its determinants. AB - AIM: To analyse sensitivity, specificity and complication rate of endoscopy, and barium enema for the detection of colorectal neoplasia. MATERIALS AND METHODS: A MEDLINE search was performed (1980-2000) directed at the endoscopic and radiologic literature on barium enema. Articles were selected based on the type of study, availability of sensitivity and specificity values in sizeable patient groups, and reports on complications. Sixty articles were included in the analysis. RESULTS: Endoscopy proved to have superior sensitivity for polyps in patients at high-risk for colorectal neoplasia. The role of endoscopy and radiology in average-risk screening populations is not known. Sensitivity and specificity rates ranged widely, probably due to bias. For the detection of small polyps endoscopy has superior performance, whereas sensitivity is similar for endoscopy and barium enema for the detection of larger (>1 cm) polyps and tumours. Overall, endoscopy is associated with a higher complication rate. CONCLUSION: Endoscopy is the preferred detection method in high-risk patients. The role of endoscopy and radiology in a screening setting requires evaluation. This review provides the test characteristics of endoscopy and radiology which are relevant for a cost-effectiveness analysis. Double-contrast barium enema may play an important role for screening purposes, owing to its good sensitivity for detecting larger (>1 cm) polyps and its lack of major complications. de Zwart, I. M.et al. (2001). Clinical Radiology56, 401-409. PMID- 11384141 TI - Dual contrast enhanced magnetic resonance imaging of the liver with superparamagnetic iron oxide followed by gadolinium for lesion detection and characterization. AB - AIM: Iron oxide contrast agents are useful for lesion detection, and extracellular gadolinium chelates are advocated for lesion characterization. We undertook a study to determine if dual contrast enhanced liver imaging with sequential use of ferumoxides particles and gadolinium (Gd)-DTPA can be performed in the same imaging protocol. MATERIALS AND METHODS: Sixteen patients underwent dual contrast magnetic resonance imaging (MRI) of the liver for evaluation of known/suspected focal lesions which included, metastases (n = 5), hepatocellular carcinoma (HCC;n = 3), cholangiocharcinoma(n = 1) and focal nodular hyperplasia (FNH;n = 3). Pre- and post-iron oxide T1-weighted gradient recalled echo (GRE) and T2-weighted fast spin echo (FSE) sequences were obtained, followed by post-Gd DTPA (0.1 mmol/kg) multi-phase dynamic T1-weighted out-of-phase GRE imaging. Images were analysed in a blinded fashion by three experts using a three-point scoring system for lesion conspicuity on pre- and post-iron oxide T1 images as well as for reader's confidence in characterizing liver lesions on post Gd-DTPA T1 images. RESULTS: No statistically significant difference in lesion conspicuity was observed on pre- and post-iron oxide T1-GRE images in this small study cohort. The presence of iron oxide did not appreciably diminish image quality of post-gadolinium sequences and did not prevent characterization of liver lesions. CONCLUSION: Our results suggest that characterization of focal liver lesion with Gd-enhanced liver MRI is still possible following iron oxide enhanced imaging.Kubaska, S.et al. (2001). Clinical Radiology, 56, 410-415 PMID- 11384142 TI - Vascularity and grey-scale sonographic features of normal cervical lymph nodes: variations with nodal size. AB - AIM: This study was undertaken to investigate variations in the vascularity and grey-scale sonographic features of cervical lymph nodes with their size. MATERIALS AND METHODS: High resolution grey-scale sonography and power Doppler sonography were performed in 1133 cervical nodes in 109 volunteers who had a sonographic examination of the neck. Standardized parameters were used in power Doppler sonography. RESULTS: About 90% of lymph nodes with a maximum transverse diameter greater than 5 mm showed vascularity and an echogenic hilus. Smaller nodes were less likely to show vascularity and an echogenic hilus. As the size of the lymph nodes increased, the intranodal blood flow velocity increased significantly (P < 0.05), whereas there was no significant variation in the vascular resistance(P> 0.05). CONCLUSIONS: The findings provide a baseline for grey-scale and power Doppler sonography of normal cervical lymph nodes. Sonologists will find varying vascularity and grey-scale appearances when encountering nodes of different sizes.Ying, M.et al. (2001). Clinical Radiology, 56, 416-419. PMID- 11384143 TI - Case report: Vesico-ureteric reflux demonstrated during barium enema examination in a patient with enterovesical fistula. PMID- 11384144 TI - Case report: Extranodal peripelvic and periureteric lymphoma--demonstration with computed tomography. PMID- 11384145 TI - Case report: Transcatheter embolization of pseudoaneurysm of the profunda femoris artery complicating fracture of the femoral neck. PMID- 11384146 TI - Case report: Hemispheric juvenile pilocytic astrocytoma with extreme exophytic growth. PMID- 11384147 TI - Case report: Caecal volvulus in association with a mesenteric dermoid. PMID- 11384148 TI - Percutaneous transhepatic lymphography. PMID- 11384149 TI - Constitutive expression and localization of COX-1 and COX-2 in rabbit iris and ciliary body. AB - Prostaglandins are involved in the regulation of intraocular pressure and the blood-aqueous barrier of the eye, and are used for the treatment of glaucoma. The decrease of the constitutively expressed PG-synthesizing enzyme cyclooxygenase-2 (COX-2) has been demonstrated in the ciliary non-pigmented epithelial layer of patients with primary open angle glaucoma. Little is known about the distribution of COX-1 and COX-2 in animals. We investigated this in the iris and ciliary body of the normal rabbit eye. The presence of COX-1 and COX-2 in freshly excised iris and ciliary body tissue from adult New Zealand White albino rabbits was demonstrated by real-time RT-PCR, and Western blot analysis. The localization of both isoforms and of the neuron-specific protein gene product 9.5 was determined by indirect immunofluorescence. Both enzymes are expressed in the iris and the ciliary body. Immunofluorescence studies including double staining techniques localized COX-1 and COX-2 to about 50% of cells in the stromal tissue of iris and ciliary body, mainly on the corneal side. They were co-localized in about 75% of these cells. Whereas all stained cells were positive for COX-1, COX-2 showed a gradient-like distribution in the stroma, with some restriction of expression near the epithelial layers, which we clearly showed to be completely negative for both COX-1 and COX-2. Also, neuronal elements did not show COX-1 or COX-2 immunoreactivity. These results establish the presence of COX-1 and COX-2 on the RNA and protein levels in normal, unstimulated rabbit iris and ciliary body. The pattern of distribution suggests a role for both enzymes in maintaining the physiology of the eye. In contrast to our results in man, non-pigmented epithelial cells of the ciliary body did not express immunoreactivity. This could account for differences in the regulation of intraocular pressure and/or blood aqueous barrier between human and rabbit eyes. PMID- 11384150 TI - Fas/Fas ligand-associated apoptosis in experimental autoimmune uveoretinitis in rodents: role of proinflammatory corticotropin-releasing hormone. AB - We have previously shown that corticotropin-releasing hormone plays an important proinflammatory role in the induction of experimental autoimmune uveoretinitis. In this study, we examined the role of apoptosis in the destruction of the retina during experimental autoimmune uveoretinitis, and the role of corticotropin releasing hormone as a local regulator of Fas and Fas Ligand expression in this condition. We evaluated apoptosis by the terminal deoxynucleotidyl transferase dUTP nick end labeling method and Fas and Fas Ligand presence by immunohistochemistry. We examined formalin-fixed, paraffin-embedded eye sections from female Lewis rats or B10.A mice immunized with the major pathogenetic epitope (R16 peptide) of the interphotoreceptor retinoid-binding protein. Female B10.A mice similarly immunized were treated with intraperitoneal injections of the rabbit anti-corticotropin-releasing hormone antibody TS-2 or nonimmune rabbit serum. The percentage of retinal cells undergoing apoptosis and the expression of Fas and Fas Ligand were increased in inflamed retinas in immunized Lewis rats and B10.A mice, compared to controls. Retinas from immunized B10.A mice treated with anti-corticotropin-releasing hormone antibody showed significantly lower apoptosis and Fas and Fas Ligand expression than placebo-treated animals. In conclusion, retinal cells in experimental autoimmune uveoretinitis undergo apoptosis associated with concurrent upregulation of Fas and Fas Ligand. The local presence of corticotropin-releasing hormone appears to be of pivotal importance in this process. PMID- 11384151 TI - Human tear fluid PDGF-BB, TNF-alpha and TGF-beta1 vs corneal haze and regeneration of corneal epithelium and subbasal nerve plexus after PRK. AB - The aim was to determine the association of tear fluid cytokine levels and post PRK corneal haze evaluated by in vivo confocal microscopy. In addition, the possible association between subbasal neural regeneration and haze formation, or epithelial regeneration were investigated. Twenty eyes of 20 patients (16 women and four men, age 30.7 +/- 7.5 years, range 21-48 years) underwent a myopic PRK. The spherical equivalent (SE) of the intended correction was -4.7 +/- 1.5 D (range -2.75 to -9.00 D). ELISA-methods were used to assess tear fluid concentrations of TGF-beta1, PDGF-BB and TNF-alpha pre-operatively, and post operatively on day 2 and at 3 months. Tear fluid flow in the collection capillary was recorded, and rates of cytokine release (= tear fluid flow-corrected concentrations) were calculated. In vivo confocal microscopy was performed at 3 months to evaluate the corneal morphology and to determine numerical haze estimate. There was wide interindividual variation between pre-operative and post operative concentrations and rates of release of TGF-beta1, PDGF-BB and TNF alpha. Subepithelial haze was observed in all corneas and the mean haze estimate was 506 +/- 401 U (100-1410 U). However, no association was found between tear fluid cytokine levels and post-PRK haze. Regenerating subbasal nerve plexus was found in 18 out of 20 corneas; in two corneas it was absent or could not be visualized due to subepithelial haze. The density of the subbasal nerve fiber bundles had a positive correlation with the epithelial thickness (Pearson correlation, r = 0.56, P = 0.011), but not with the haze estimate or the thickness of the haze area. At 3 months post-PRK, haze could be observed in all patients. The results suggest that tear fluid cytokine analysis, as measured, may not be suitable for screening the potential candidates for haze formation. We did not find any correlation between haze and regeneration of subbasal nerve plexus, but we demonstrated that the regeneration of subbasal nerve plexus might have significant influence on regulation of epithelial healing. PMID- 11384152 TI - Suppressive effects of thyroxine on glucocorticoid (gc)-induced metabolic changes and cataract formation on developing chick embryos. AB - To prevent cataracts induced by glucocorticoids (GC) as a systemic disease, the suppression of oxidative stress caused by GC in the hepatic metabolism is of significant interest. In this study, to elucidate the formative mechanism of GC induced cataracts, we examined the preventive effect and then analysed the mechanisms of thyroxine on GC-induced cataract formation. Fifteen day old chick embryos were administered with 0.25 micromol hydrocortisone succinate sodium (HC), and then 12-30 nmol of thyroxine 4 hr after HC administration. At the indicated time after HC treatment, we examined the incidence of cataract formation, the levels of serum glucose and lipids, lenticular and hepatic glutathione (GSH), and lipid peroxide (LPO) in the lens, blood and liver. Almost all lenses (96%) removed 48 hr after HC administration were opaque. Thyroxine prevented HC-induced cataract formation effectively, and suppressed the elevations of serum glucose and LPO in the lens, blood and liver. The treatment prevented the decreased lenticular GSH level at 48 hr, but the hepatic GSH level at 24 hr remained lowered in contrast to the results of previous studies using insulin. Moreover, thyroxine did not decrease the elevated serum lipid level (triglyceride and non-esterified fatty acid) caused by HC. Under thyroxine treatment, in constant to insulin, acceleration of GSH-GSSG cycle rather than GSH de novo synthesis keeps a certain level of hepatic GSH necessary for diminishing the elevation of LPO as a risk factor of GC-induced cataract formation. The regulation of metabolic changes ensured the maintenance of hepatic GSH level, which is necessary to reduce oxidative stress produced by GC and to protect the lens from oxidative stress leading to opacification. PMID- 11384153 TI - Tgfbeta receptor expression in lens: implications for differentiation and cataractogenesis. AB - TGFbeta induces changes characteristic of some forms of cataract. However, the responsiveness of lens epithelial cells to TGFbeta is age-dependent; weanling and adult, but not neonatal, lens epithelial cells respond. This study investigated TGFbeta receptor (TbetaRI and TbetaRII) expression during rat lens development and the effects of FGF-2 on TGFbeta responsiveness and TbetaR expression. Immunofluorescence, immunoblotting, RT-PCR and in situ hybridization were used to examine the spatio-temporal expression patterns of TbetaR. Lens explants were used to investigate the effects of FGF-2 on TGFbeta responsiveness and TbetaR expression. In the lens epithelium, little or no immunoreactivity was detected at P3 but at P21 there was distinct reactivity for TbetaRI and TbetaRII. Reactivity for both receptors was also found in the differentiating fibers in the transitional zone and cortex at both ages. Western blotting of lens membrane extracts identified multiple molecular weight forms of TbetaRI (30, 50, 90 kDa) and TbetaRII (70-120 kDa). In situ hybridization with a rat probe for Alk5 (TbetaRI) showed that the lens expresses Alk5 mRNA in epithelium and fibers throughout development. A rat TbetaRII probe revealed distinct expression of a TbetaRII mRNA in lens fibers throughout development and in the lens epithelium at P21 but not at P3. In vitro studies showed that lens epithelial explants from P9 rats did not undergo cataractous changes in response to TGFbeta but P13 explants did. Addition of FGF-2 to P9 explants induced increased TbetaR immunoreactivity and enhanced the competency of lens epithelial cells to respond to TGFbeta. These data indicate that the overall increased expression of TGFbeta receptors in lens epithelium during postnatal development (P3-P21) underlies an age-related change in TGFbeta responsiveness. The results also suggest that lens cells may express multiple forms of TbetaR. Expression of TbetaR in lens fibers throughout lens development and the induction of enhanced TbetaR expression by FGF suggest a role for TGFbeta signaling during FGF-induced responses and fiber differentiation. PMID- 11384154 TI - Glutathione transport in human retinal pigment epithelial (HRPE) cells: apical localization of sodium-dependent gsh transport. AB - The study was undertaken to identify and localize GSH transport in non transformed cultured human retinal pigmented epithelial cells (HRPE). In confluent monolayers exhibiting high transepithelial resistance (TER 700-1000 Omega cm(-2)), apical and basolateral GSH uptake were determined after introducing(35)S-GSH (+ 1 m M GSH) to the apical side or basal side in NaCl (Na+ containing) or choline chloride (Na+ -free) buffers. Cells in growth medium or in incubation buffers were pretreated with acivicin to inhibit gamma glutamyltranspeptidase (GGT). GSH efflux was measured after labelling the intracellular GSH pool by incubation overnight with 35 S-cysteine and quantitating the release of labelled GSH into the medium. Uptake of GSH was found at both the apical and basolateral membranes of HRPE cells. Inhibition of gamma glutamyltranspeptidase (GGT) with acivicin did not alter mean GSH uptake (nmol per million cells per 30 min) significantly at the apical (1.63 +/- 0.32 vs 1.45 +/- 0.30; with and without acivicin respectively) or the basolateral (1.17 +/- 0.21 vs 1.44 +/- 0.38) membranes. Transport was verified to be in the form of intact GSH by HPLC. Uptake was unaffected by the removal of Na+ at the basolateral membrane while apical uptake exhibited partial but significant (approximately 40%) Na+ -dependency. Net GSH efflux (nmol per million cells per min) to the apical side of HRPE cells was higher than to the basolateral side in the presence of sodium. Transepithelial flux in the basolateral to apical direction was approximately 17-fold higher than the apical to basolateral direction resulting in a net flux of GSH to the apical side. In conclusion, HRPE cells exhibit GSH transport by Na+ -dependent and Na+ -independent mechanisms. The Na+ -dependent GSH transporter is localized to the apical membrane of HRPE cells. PMID- 11384155 TI - Electroretinographic abnormalities in a rat glaucoma model with chronic elevated intraocular pressure. AB - The purpose of this study was to determine the ability of electroretinographic (ERG) measurements to document progression of the retinopathy in a rat glaucoma model. Thirty four rats with a chronic intraocular pressure (IOP) elevation induced in one eye by cautery of three episcleral/extra-orbital veins were studied in four separate groups. ERGs were recorded sequentially in Group A rats (n = 12) at baseline, and after approximately 20, 40 and 60 days of high IOP, and in three additional groups of rats (n = 6 or 10 per group) after approximately 58, 30 and 175 days of high IOP, respectively. Scotopic ERG parameters recorded simultaneously from both eyes in Group A rats were: a- and b-wave amplitudes, implicit times, oscillatory potential amplitudes (OPs) determined at three different light-flash intensities, and the light-adapted (photopic) ERG b-wave amplitude. In the other groups of rats, only scotopic ERG a-wave, b-wave and OP amplitudes were measured.In Group A rats that were followed sequentially, all the ERG parameters recorded with attenuated stimuli showed significant time-dependent changes in glaucomatous eyes relative to their contralateral normal eyes, with OPs showing the earliest significant difference after only 3 weeks of high IOP. When different groups of unilateral glaucomatous rats were compared beyond 8 weeks of elevated IOP only the OPs showed a continued decrease with time and good discrimination between glaucoma and normal eyes. Over a 25 week period of high IOP the scotopic OPs measured with attenuated light stimuli declined at the rate of approximately 1.5% per week and provided the best ERG measure to monitor progression of retinal pathophysiology in the vein-occlusion rat glaucoma model. PMID- 11384156 TI - Smad translocation and growth suppression in lens epithelial cells by endogenous TGFbeta2 during wound repair. AB - To determine whether endogenous TGFbeta affects lens epithelial cells during repair after an anterior capsule injury in mice, we studied translocation of Smad proteins, which carry the TGFbeta signal from cell surface receptors to promoters in nuclei. We immunolocalized Smads in murine lenses at intervals up to 8 weeks following capsular injury. Effects of injecting TGFbeta neutralizing antibodies on Smad4 location and cell proliferation were examined at 24 hr after injury. Finally, we examined whether exogenous TGFbeta2 induced Smad nuclear translocation in murine lenses in organ culture. Cell proliferation was quantitated by 5-bromo-2'-deoxyuridine (BrdU) labelling. In uninjured lenses, Smads were located in the cytoplasm. In injured lenses, nuclear localization of Smads was observed in cells next to the capsular break from 8 to 24 hr after the injury, and was observed peripheral to the break at 48 hr. Nuclear Smads then continued to be observed occasionally in a minority of cells. Injection of antibodies neutralizing TGFbeta2, but not TGFbeta1 or TGFbeta3, inhibited Smad4 nuclear translocation and resulted in the appearance of BrdU-positive anterior epithelial cells. With the lenses in culture, transient nuclear localization of Smads occurred between 3 and 24 hr in response to continuous exposure to TGFbeta2. No nuclear translocation was seen at 48 hr. Endogenous TGFbeta2 affects lens cells during wound repair after anterior capsule injury, inhibiting lens cell proliferation during the early phase. Nuclear translocation of Smads in lens epithelial cells is transient even with continuous exposure to TGFbeta2. PMID- 11384157 TI - Is there a glucocorticoid receptor in the bovine lens? AB - Prolonged glucocorticoid therapy is a risk factor for cataract development. The mechanism remains unknown. If cataract results from the direct effect of steroids on lens function, a glucocorticoid receptor is required. In order to determine whether such a receptor was present in the bovine lens, metabolic and steroid binding experiments were undertaken. Cultured bovine lens epithelial cells were exposed to 10(- 4)and 10(-8) M dexamethasone or prednisolone and the uptake and incorporation of(14)C leucine,(14)C glucose and(3)H thymidine, examined. Neither glucocorticoid affected cell protein synthesis or glucose uptake. Both dexamethasone concentrations and the lower concentration of prednisolone had no effect on thymidine uptake or incorporation, however, the 10(-4) M prednisolone exposure reduced these by 15 +/- 5%. This regulation is thought to be due to membrane fluidity changes and not the action of the glucocorticoid receptor. As the glucocorticoid receptor is very heat labile in vitro, the effects of increasing temperature on dexamethasone binding by proteins from lens epithelium, lens nucleus and liver were examined. At 0 degree C, lens epithelial extract bound nine-fold more dexamethasone than liver extract. After exposure to 37 degrees C, liver binding decreased by 66% whereas that for lens epithelium increased by 18%. For both lens extracts, steroid binding increased with temperature up to 50 degrees C. Scatchard analysis of the steroid binding kinetics showed there to be no high affinity sites in lens epithelial extract, with the binding best described as a non-specific partitioning event. Western blotting with a specific glucocorticoid receptor antibody revealed protein bands of approximately 94 and 79 kDa in liver, which is known to contain significant levels of receptor. No immunoreactivity was observed for lens epithelial extract. Therefore, within the limits of detection, these results suggest the bovine lens does not contain a glucocorticoid receptor. This raises questions about the validity of receptor-mediated mechanisms proposed for cataract development. PMID- 11384158 TI - Mislocalization of melanosomal proteins in melanocytes from mice with oculocutaneous albinism type 2. AB - More than 10% of admissions worldwide to institutions for the visually impaired are due to some form of albinism. The most common form, oculocutaneous albinism type 2, results from mutations at the p locus. The function of the p gene is yet to be determined. It has been shown that melanocytes from p -null mice exhibit an abnormal melanosomal ultrastructure in addition to alterations in activity and localization of tyrosinase, a critical melanogenic enzyme. In light of these observations, we examined tyrosinase trafficking in p -null vs wildtype mouse melanocytes in order to explore p function. Electron microscopy of wildtype melan a and p -null melan-p1 cells demonstrated accumulation of tyrosinase in 50 nm vesicles throughout the cell in the absence of p, an observation corroborated by an increase in tyrosinase activity in vesicle-enriched fractions from melan-p1 compared to melan-a cells. Misrouting in the absence of p was not limited to tyrosinase; a second melanosomal protein, tyrosinase-related protein 1, also trafficked incorrectly. In melan-p1, mislocalization led to secretion of tyrosinase into the medium. Adding tyrosine to the medium was found to partially correct tyrosinase trafficking and to reduce secretion; the cysteine protease inhibitor E64 also reduced secretion. We propose that p is required by melanocytes for transport of melanosomal proteins. In its absence, tyrosinase accumulates in vesicles and, in cultured melanocytes, is proteolysed and secreted. PMID- 11384159 TI - Human trabecular meshwork cells express the ciliary neurotrophic factor (cntf) tripartite receptor complex. PMID- 11384160 TI - Increased Retinal Lipid Peroxidation in Early Diabetes is not Associated with Ascorbate Depletion or Changes in Ascorbate Redox State. PMID- 11384161 TI - Effect of an electric field on the motility of Entamoeba histolytica examined by multiple-beam interference microscopy. AB - A recently developed multiple-beam interference microscopic technique has been used to visualize submicroscopic structures of Entamoeba histolytica and their movements in applied external electric fields. The movements were videorecorded and it was found that at low current (120 microA) pseudopods are filled with hyaline ectoplasm. At slightly higher current (about 150 microA), the amoeba stops extending the pseudopods and loosens its attachment to the surface. At higher currents (200 microA), it forms a cyst and remains immobile for a time. Before this stage is reached a narrow ring is formed around the nucleus due to alterations in the proteins to protect it. PMID- 11384162 TI - Clonorchis sinensis: molecular cloning and characterization of 28-kDa glutathione S-transferase. AB - A 28-kDa glutathione S-transferase (Cs28GST) was purified from a Clonorchis sinensis cytosolic fraction through anion-exchange and glutathione-affinity column chromatographies. A monoclonal antibody raised against Cs28GST reacted specifically to the C. sinensis antigen among trematode proteins. A putative peptide of 212 amino residues deduced from a cDNA clone appeared homologous with 28-kDa GST of trematodes, and its secondary structural elements predicted a GSH binding site. Recombinant Cs28GST showed GST enzyme activity with CDNB substrate and was sensitive to the model inhibitors. The recombinant Cs28GST was antigenically indistinguishable from the native form and was recognized specifically by C. sinensis-infected human sera. The Cs28GST was localized in the tegument and underlying mesenchymal tissues. It is suggested that Cs28GST may play significant physiological roles against bioreactive molecules and be a useful reagent for serodiagnosis of clonorchiasis. PMID- 11384163 TI - Trypanosoma cruzi: exogenously regulated gene expression. AB - A regulated expression vector would provide a strong tool for the dissection of gene function in Trypanosoma cruzi. Herein, we establish a system in which genes in T. cruzi expression vectors can be exogenously regulated by tetracycline. We first generated strains of T. cruzi that stably express the repressor of the bacterial tetracycline resistance gene and T7 RNA polymerase. Based on these strains, we developed two T. cruzi expression systems regulated by tetracycline- the first by use of a regulated rRNA promoter and the second by use of a regulated T7 promoter. In the former, we constructed an expression vector in which tetracycline resistance gene operators flank the transcription start point of the T. cruzi rRNA gene promoter. Reporter gene activity from this modified promoter was regulated up to 20-fold in the presence of different concentrations of tetracycline. In the T7 system, tetracycline resistance gene operators flank the transcription start point of the T7 promoter. Reporter gene activity from this modified promoter was regulated up to 150-fold in the presence of different concentrations of tetracycline. Expression in these systems was repressed when tetracycline was removed even after full induction for extended periods in the presence of tetracycline. We are now using these two systems to test protein function in T. cruzi. PMID- 11384164 TI - Nitric oxide synthase in filariae: demonstration of nitric oxide production by embryos in Brugia malayi and Acanthocheilonema viteae. AB - The radical gas nitric oxide (NO) is synthesized by nitric oxide synthase (NOS) from l-arginine and molecular oxygen. Nitric oxide is an important signaling molecule in invertebrate and vertebrate systems. Previously we have shown that NOS is localized to more tissues in Brugia malayi than has been reported in Ascaris suum. In this paper, we analyze the distribution of NOS in Acanthocheilonema viteae, a filarial nematode that differs from B. malayi in that A. viteae females release microfilariae without a sheath. A. viteae is also one of a few filarial parasites without the Wolbachia intracellular endosymbiont. By use of a specific antibody, NOS was demonstrated in extracts of A. viteae and Dirofilaria immitis. The localization pattern of NOS in A. viteae was similar to that seen in B. malayi, with the enzyme localized to the body wall muscles of both sexes, developing spermatozoa, intrauterine sperm, and early embryos. By use of DAF-2, a fluorescent indicator specific for nitric oxide, the embryos of B. malayi and A. viteae were demonstrated to produce NO ex utero. The near identical staining patterns seen in A. viteae and B. malayi argue that NO is not produced by Wolbachia, nor is it produced by the nematodes in response to the infection. Localization of NOS to the sperm of filarial nematodes suggests a role for NO during fertilization as has been described for sea urchin and ascidian fertilization. Demonstration of the activity of embryonic NOS supports our earlier hypothesis that NO is a signaling molecule during embryogenesis in filarial nematodes. PMID- 11384165 TI - Echinococcus granulosus: Cloning and Functional in Vitro Characterization of an Actin Filament Fragmenting Protein. AB - We report the isolation and characterization of an Echinococcus granulosus gene that codes for a protein with actin filament fragmenting and nucleating activities (EgAFFP). The genomic region corresponding to the EgAFFP gene presents a coding sequence of 1110 bp that is interrupted by eight introns. The EgAFFP deduced amino acid sequence is about 40% homologous to those of several members of the gelsolin family, such as Physarum polycephalum fragmin, Dictyostelium discoideum severin, and Lumbricus terrestris actin modulator. As do other proteins of the same family, EgAFFP presents three repeated domains, each one characterized by internal conserved amino acid motifs. Assays with fluorescence labeled actin showed that the full-length recombinant EgAFFP effectively binds actin monomers in both a calcium-dependent and calcium-independent manner and also presents actin nucleating and severing activities. PMID- 11384167 TI - Electron spin relaxation via vibronic level of rhodium(II) hexacyanide complex in KCl crystal. AB - Electron spin-lattice relaxation rates for the low-spin [Rh(CN)(6)](4-) complex in KCl were measured by the inversion recovery and saturation recovery techniques, in the range of 5 to 30 K. Angular variation experiments indicate that electron spin-lattice relaxation times present axial symmetry. The data fit very well to a relaxation process involving localized anharmonic vibration modes, also responsible for the g tensor temperature dependence. PMID- 11384166 TI - Identification of Trypanosoma cruzi genotypes circulating in Chilean chagasic patients. AB - Parasite DNA amplified by PCR from blood of 73 chagasic children and adults of two endemic areas of Chile were studied by Southern blot and/or dot blot hybridization analysis with a panel of three minicircle probes corresponding to the parasite genotypes (clonets 19, 33 and 39). The hybridization pattern of the PCR positive samples identified clonets 39, 19/20, and 32/33 with frequencies of 0.84, 0.32 and 0.26, respectively. A total of 31 samples corresponded to mixed infections. The most frequently found mixtures were: clonets 39 and 19/20 (14 cases), followed by clonets 39 and 32/33 (8 cases), clonets 39, 32/33 and 19/20 (8 cases), and clonets 32/33 and 19/20 (1 case). Amplified DNA from 9 cases showed no hybridization signal with none of the three studied probes indicating that other genotypes different to the ones mentioned are circulating in humans, but that the clonets used as probes are the most prevalent ones in terms of transmission in the endemic areas studied. A biological characterization of 34 T. cruzi populations isolated from the xenodiagnosis of the patients was performed on an experimental murine model. The biochemical characterization of the parasite populations by molecular karyotype determined that the most frequent parasite isolated from patients belongs to clonet 39. PMID- 11384168 TI - Selective storage of magnetization in strongly relaxing spin systems. AB - We have investigated the use of a "split-sinc" RF pulse to selectively store magnetization from a selected region of a sample, for later recall in imaging or spectroscopy experiments. The pulse sequence is based on an original suggestion by Post et al. (West German Patent No. P3209263.6, 13 March 1982), later implemented by Aue et al. (J. Magn. Reson. 56, 350 (1984)). We have carried out detailed numerical calculations using the Bloch equations and show that this particular sequence is robust in the face of strong transverse relaxation, and we demonstrate its application to imaging of polymer samples in shearing and extensional flow cells. PMID- 11384169 TI - Optimization of residual water signal removal by HLSVD on simulated short echo time proton MR spectra of the human brain. AB - Suppression of the residual water signal from proton magnetic resonance (MR) spectra recorded in human brain is a prerequisite to an accurate quantification of cerebral metabolites. Several postacquisition methods of residual water signal suppression have been reported but none of them provide a complete elimination of the residual water signal, thereby preventing reliable quantification of brain metabolites. In the present study, the elimination of the residual water signal by the Hankel Lanczos singular value decomposition method has been evaluated and optimized to provide fast automated processing of spectra. Model free induction decays, reproducing the proton signal acquired in human brain localized MR spectroscopy at short echo times (e.g., 20 ms), have been generated. The optimal parameters in terms of number of components and dimension of the Hankel data matrix allowing complete elimination of the residual water signal are reported. PMID- 11384170 TI - Convection compensated electrophoretic NMR. AB - A novel method of convection compensated ENMR (CC-ENMR) has been developed to detect electrophoretic motion of ionic species in the presence of bulk solution convection. This was accomplished using a gradient moment nulling technique to remove spectral artifacts from heat-induced convection and using the polarity switch of the applied electric field to retain spin phase modulations due to electrophoretic flow. Experiments were carried out with a mixture of 100 mM L aspartic acid and 100 mM 4,9-dioxa-1,12-dodecanediamine to demonstrate this new method of ENMR. CC-ENMR enhances our previously developed capillary array ENMR (CA-ENMR) in solving the convection problem. The combined CA- and CC-ENMR approach strengthens the potential of multidimensional ENMR in simultaneous structural determination of coexisting proteins and protein conformations in biological buffer solutions of high ionic strength. Structural mapping of interacting proteins during biochemical reactions becomes possible in the future using ENMR techniques, which may have a profound impact on the understanding of biological events, including protein folding, genetic control, and signal transduction in general. PMID- 11384171 TI - Solid state proton imaging detected by quadrupole resonance. AB - A double resonance method for imaging of solid materials containing quadrupole nuclei via the coupled protons is reported. The technique uses a static field gradient to encode the position on the protons and the method of double resonance spin-echo to detect the occurrence of proton resonances by affecting the zero field echo signal from the quadrupole system. The double resonance imaging method offers the advantages of higher spatial resolution and straightforward image reconstruction for powder samples compared with rotating-frame and Zeeman perturbated nuclear quadrupole resonance encoding techniques. PMID- 11384172 TI - Separating structure and dynamics in CSA/DD cross-correlated relaxation: a case study on trehalose and ubiquitin. AB - We present two new sensitivity enhanced gradient NMR experiments for measuring interference effects between chemical shift anisotropy (CSA) and dipolar coupling interactions in a scalar coupled two-spin system in both the laboratory and rotating frames. We apply these methods for quantitative measurement of longitudinal and transverse cross-correlation rates involving interference of (13)C CSA and (13)C-(1)H dipolar coupling in a disaccharide, alpha,alpha-D trehalose, at natural abundance of (13)C as well as interference of amide (15)N CSA and (15)N-(1)H dipolar coupling in uniformly (15)N-labeled ubiquitin. We demonstrate that the standard heteronuclear T(1), T(2), and steady-state NOE autocorrelation experiments augmented by cross-correlation measurements provide sufficient experimental data to quantitatively separate the structural and dynamic contributions to these relaxation rates when the simplifying assumptions of isotropic overall tumbling and an axially symmetric chemical shift tensor are valid. PMID- 11384173 TI - Imaging the long-range dipolar field in structured liquid state samples. AB - We describe imaging experiments in which the pattern of the dipolar field generated by spatially modulated nuclear magnetization is directly visualized in simply structured phantoms. Two types of experiment have been carried out at 11.7 T using (1)H NMR signals. In the first, the field from a single spin species is imaged via its own NMR signal. In the second, the NMR signal from one spin species is used to image the field generated by a second species. The field patterns measured in these experiments correspond well with those calculated using simple theoretical expressions for the dipolar field. The results also directly demonstrate the spatial sensitivity of the signal generated using dipolar field effects, indicating that the range of the field depends upon the inverse of the spatial frequency with which the magnetization is modulated. PMID- 11384174 TI - Relaxation behavior of laser-polarized (129)Xe gas: size dependency and wall effect of the T(1) relaxation time in glass and gelatin bulbs. AB - Size dependency of the relaxation time T(1) was measured for laser-polarized (129)Xe gas encapsulated in different sized cavities made by glass bulbs or gelatin capsules. The use of laser-polarized gas enhances the sensitivity a great deal, making it possible to measure the longer (129)Xe relaxation time in quite a short time. The size dependency is analyzed on the basis of the kinetic theory of gases and a relationship is derived in which the relaxation rate is connected with the square inverse of the diameter of the cavity. Such an analysis provides a novel parameter which denotes the wall effect on the relaxation rate when a gas molecule collides with the surface once in a second. The relaxation time of (129)Xe gas is also dependent on the material which forms the cavity. This dependency is large and the relaxation study using polarized (129)Xe gas is expected to offer important information about the state of the matter of the cavity wall. PMID- 11384175 TI - Development of NMR instrumentation to achieve excitation of large bandwidths in high-resolution spectra at high field. AB - A prototype 2.5-mm (1)H high-resolution probe for an 18.8-T (800 MHz) nuclear magnetic resonance spectrometer has been designed, together with a dedicated amplifier capable of delivering up to 1 kW of power. This probe permits a 90 degrees pulse length of 2 mus to be achieved at 300 W, corresponding to an excitation bandwidth of +/-125 kHz. Probe performances were tested on samples commonly used for this purpose as well as on protein and paramagnetic model compound samples. It is shown that this probe is useful for a wide range of applications at high magnetic field, especially in the study of systems characterized by very broad and far-shifted resonances and in experiments that require high-power radiofrequency irradiation. PMID- 11384176 TI - Probing proteins in solution by (129)Xe NMR spectroscopy. AB - The interaction of xenon with different proteins in aqueous solution is investigated by (129)Xe NMR spectroscopy. Chemical shifts are measured in horse metmyoglobin, hen egg white lysozyme, and horse cytochrome c solutions as a function of xenon concentration. In these systems, xenon is in fast exchange between all possible environments. The results suggest that nonspecific interactions exist between xenon and the protein exteriors and the data are analyzed in term of parameters which characterize the protein surfaces. The experimental data for horse metmyoglobin are interpreted using a model in which xenon forms a 1:1 complex with the protein and the chemical shift of the complexed xenon is reported (Locci et al., Keystone Symposia "Frontiers of NMR in Molecular Biology VI", Jan. 9--15, 1999, Breckenridge, CO, Abstract E216, p. 53; Locci et al., XeMAT 2000 "Optical Polarization and Xenon NMR of Materials", June 28--30, 2000, Sestri Levante, Italy, p. 46). PMID- 11384177 TI - A new two-dimensional pulse sequence for T(2)* measurements of protons in (13)C isotopomers. AB - A new two-dimensional pulse sequence for T(2)* measurement of protons directly coupled to (13)C spins is proposed. The sequence measures the tranverse relaxation time of heteronuclear proton single-quantum coherence under conditions of free precession and is therefore well suited to evaluate relaxation losses of proton magnetization during preparation delays of heteronuclear pulse experiments in analytical NMR. The relevant part of the pulse sequence can be inserted as a "building block" into any direct or inverse detecting H,C correlation pulse sequence if proton spin-spin relaxation is to be investigated. In this contribution, the building block is inserted into a HETCOR as well as into a HMQC pulse sequence. Experimental results for the HETCOR-based sequence are given. PMID- 11384178 TI - Effects of higher rank multipoles on relaxation of an I = 3 spin system. AB - Magnetic multipoles of rank higher than one become active in spin systems with I > 1/2 and their contribution to relaxation depends on dynamics. The appearance of multipole terms complicates the relaxation description and supports the multiexponential behavior of relaxation. In this paper the effects of high-rank multipoles on lineshape and longitudinal relaxation of I = 3 spin systems are presented. Results obtained from both numerical simulation and experimental data show that longitudinal and transverse relaxation are strongly influenced by these multipole terms, especially at lower temperature where, due to lower molecular mobility, the extreme narrowing condition is not fulfilled. PMID- 11384179 TI - Dipolar and J encoded DQ MAS spectra under rotational resonance. AB - A two-dimensional (2D) double-quantum (DQ) experiment under rotational resonance (R(2)) conditions is introduced for evaluating dipolar couplings in rotating solids. The contributions from the R(2)-recoupled dipolar interaction and the J coupling can be conveniently separated in the resulting 2D R(2)-DQ spectrum, so that the unknown dipolar coupling can readily be extracted, provided that the values of the involved J coupling constants are known. Since the measured parameters are integral intensity ratios between suitably chosen absorption peaks in the 2D spectrum, the proposed method is characterized by a reduced sensitivity to relaxation parameters. The effect of rotor-modulated terms, including chemical shift anisotropy, is efficiently averaged out by synchronizing the excitation/reconversion time with the rotor period. All of these features are demonstrated theoretically by the example of two model systems, namely, isolated spin-pairs and a three-spin system. The results of the theoretical models are applied to both (13)C and (1)H nuclei to extract dipolar couplings in uniformly (13)C labeled L-alanine and a crosslinked natural rubber. PMID- 11384180 TI - Longitudinally detected ESR magnetometer for wide-range measurements of low fields. AB - A new magnetometer utilizing a longitudinally detected ESR (LODESR) method was developed. The probe head of the LODESR magnetometer is equipped with a single turn coil (8 mm in diameter) which has a very wide bandwidth because the reactance of the coil is always smaller than the resistance of the transmission line (50 ohm) at frequencies less than 700 MHz. Thus, an absolute magnetic field could be measured over a wide range (2 to 9 mT) using this magnetometer without changing the probe head. PMID- 11384181 TI - Solid-state NMR spectroscopy of paramagnetic metallocenes. AB - The paramagnetic metallocenes and decamethylmetallocenes (C(5)H(5))(2)M and (C(5)Me(5))(2)M with M=V (S=3/2), Mn (S=5/2 or 1/2), Co (S=1/2), and Ni (S=1) were studied by (1)H and (13)C solid-state MAS NMR spectroscopy. Near room temperature spinning sideband manifolds cover ranges of up to 1100 and 3500 ppm, and isotropic signal shifts appear between -260 and 300 ppm and between -600 and 1640 ppm for (1)H and (13)C NMR spectra, respectively. The isotropic paramagnetic signal shifts, which are related to the spin densities in the s orbital of ligand atoms, were discussed. A Herzfeld--Berger spinning sideband analysis of the ring carbon signals yielded the principal values of the paramagnetic shift tensors, and for metallocenes with a small g-factor anisotropy the electron spin density in the ligand pi system was determined from the chemical shift anisotropy. The unusual features of the (1)H and (13)C solid-state NMR spectra of manganocene were related to its chain structure while temperature-dependent (1)H MAS NMR studies reflected antiferromagnetic interaction between the spin centers. PMID- 11384182 TI - One micrometer resolution NMR microscopy. AB - We obtained a magnetic resonance image of 1 microm resolution and 75 microm(3) voxel volume for a phantom filled with hydrocarbon oil within an hour at 14.1 T. For this work, a specially designed probe with a high sensitivity RF coil and gradient coils generating over 1000 G/cm was built. The optimal pulse sequence was analyzed in consideration of the bandwidth, diffusion coefficients, and T(1) and T(2) relaxations of the medium. The system was applied to the in vivo imaging of a geranium leaf stem to get the images of 2 microm resolution and 200 microm(3) voxel volume. PMID- 11384183 TI - Nuclear relaxation time images by radiofrequency field gradients applied to the study of solvent permeation into polymeric materials. AB - T(2) images are obtained by two interleaved B(1)-gradient imaging experiments preceded by CPMG trains of different lengths. The method is assessed by means of a phantom involving compartments of different, though relatively close, T(2) values. T(1) images arise from a previously published procedure also based on two interleaved B(1)-gradient imaging experiments involving different evolution of the longitudinal magnetization. Both types of image appear to be useful in view of the structural characterization of polymer samples through the T(2) and T(1) distribution of a solvent embedded in the material. PMID- 11384185 TI - Rate constants determined by nuclear magnetic resonance. AB - Fast kinetic methods are used to measure reactions that take place in less time than required to mix the reagents manually and to measure the reaction by usual methods, like UV-visible spectrophotometry and fluorescence. The best known of them are rapid-mixing and relaxation methods, which are used for reactions with half-times in the millisecond and microsecond ranges, respectively. The picosecond range is usually measured with electrical field and ultrasonic waves (A. Cornish-Bowden, 1976, Principles of Enzyme Kinetics, pp. 164-167, Butterworths, London). Normally these very fast rates occur when a ligand binds to or dissociates from a protein. When the binding is mediated only by the diffusion, the lower limit of the association rate constant (k(on)) should not exceed the value of a diffusion-controlled reaction (around 10(10) M(-1) s(-1)). Therefore, the values most frequently found for these rate constants, for example, in the association of a substrate with an enzyme, are in the range 10(6) to 10(9) M(-1) s(-1) (M. Eigen and G. G. Hammes, 1963, Adv. Enzymol. 25, 1-38). The values for the dissociation rate constants (k(off)) for these reactions, which depend on the equilibrium constant for the enzyme-substrate complex interaction, are in the range 10(1) to 10(5) s(-1), most often between 10(3) and 10(4) s(-1) (A. Fersht, 1999, Structure and Mechanism in Protein Science, pp. 164 165, Freeman, New York). If the equilibrium constant is known, and the value of koff is determined by nuclear magnetic resonance (NMR), as described in this chapter, the value of k(on) can be calculated; this should not exceed the value of diffusion rate in the media in which the reaction is performed. PMID- 11384186 TI - Dual-color fluorescence cross-correlation spectroscopy for monitoring the kinetics of enzyme-catalyzed reactions. AB - Dual-color fluorescence correlation spectroscopy is a biophysical technique that enables precise and sensitive analyzes of molecular interactions. It is unique in its ability to analyze reactions in real time at nanomolar substrate concentrations and below, especially when applied to the monitoring of enzyme catalyzed reactions. Furthermore, it offers a wide range of accessible reactions, restricted only by the prerequisite that a chemical bond or a physical interaction between two spectrally distinguishable fluorophores is established or broken. Recently, the optical setup of dual-color fluorescence correlation spectroscopy has been extended toward two-photon excitation, resulting in several advantages compared with standard excitation, such as lower fluorescence background, an even larger spectrum of potential fluorescence dyes to be used, as well as a more stable and simplified optical setup. So far, the method has been successfully employed to analyze the kinetics of nucleic acid and peptide modifications catalyzed by nucleases, polymerases, and proteases. PMID- 11384187 TI - Uses of isotope effects in the study of enzymes. AB - There have been few recent additions to the technical methods employed in the study of isotope effects, notable exceptions being the use of high pressure as an experimental variable and the measurement of heavy-atom isotope effects on maximal velocities using continuous-flow techniques. Most of the innovations are in the realm of new experimental designs that allow the asking of new questions. These designs include the use of isotope effects to: determine kinetic mechanisms, distinguish between changes in enzymatic activity and loss of active enzyme, distinguish between reactant-state origins and transition-state origins and quantify hydrogen tunneling, separate and quantify multiple origins of solvent isotope effects, distinguish between concerted and stepwise chemical mechanisms, characterize bond order changes in ligand binding, distinguish different pathways of inhibitor binding, and estimate intrinsic isotope effects. PMID- 11384188 TI - Trapping reaction intermediates in macromolecular crystals for structural analyses. AB - The development of "time-resolved" crystallographic methods, including trapping of reaction intermediates and rapid data collection, allows the comparative study of discrete structural species formed during a macromolecular reaction, such as enzymatic catalysis, ribozyme cleavage, or a protein photocycle. The primary technical details that must be addressed in such studies are the reaction initiation, the accumulation of a specific reaction species throughout the crystal, the lifetime of that species and of the crystal under the experimental conditions, and the method used to collect X-ray data. Methods of reaction initiation range from substrate diffusion, which is appropriate for the visualization of very long-lived intermediates, to photolysis, which is appropriate for the accumulation of rate-limited species with half-lives ranging from milliseconds to nanoseconds. This review discusses various methods for initiating turnover in crystals and trapping rate-limiting species for structural studies. PMID- 11384189 TI - Photochemically induced electron transfer. AB - Biochemical reactions involving electron transfer between substrates or enzyme cofactors are both common and physiologically important; they have been studied by means of a variety of techniques. In this paper we review the application of photochemical methods to the study of intramolecular electron transfer in hemoproteins, thus selecting a small, well-defined sector of this otherwise enormous field. Photoexcitation of the heme populates short-lived excited states which decay by thermal conversion and do not usually transfer electrons, even when a suitable electron acceptor is readily available, e.g., in the form of a second oxidized heme group in the same protein; because of this, the experimental setup demands some manipulation of the hemoprotein. In this paper we review three approaches that have been studied in detail: (i) the covalent conjugation to the protein moiety of an organic ruthenium complex, which serves as the photoexcitable electron donor (in this case the heme acts as the electron acceptor); (ii) the replacement of the heme group with a phosphorescent metal substituted porphyrin, which on photoexcitation populates long-lived excited states, capable of acting as electron donors (clearly the protein must contain some other cofactor acting as the electron acceptor, most often a second heme group in the oxidized state); (iii) the combination of the reduced heme with CO (the photochemical breakdown of the iron-CO bond yields transiently the ground state reduced heme which is able to transfer one electron (or a fraction of it) to an oxidized electron acceptor in the protein; this method uses a "mixed valence hybrid" state of the redox active hemoprotein and has the great advantage of populating on photoexcitation an electron donor at physiological redox potential). PMID- 11384190 TI - Kinetic and titration methods for determination of active site contents of enzyme and catalytic antibody preparations. AB - Kinetic characterization of enzymes and analogous catalysts such as catalytic antibodies requires knowledge of the molarity of functional sites. Various stoichiometric titration methods are available for the determination of active site concentrations of some enzymes and these are exemplified in the second part of this article. Most of these are not general in that they require the existence of certain types of either intermediate or active-site residues that are susceptible to specific covalent modification. Thus they are not readily applicable to many enzymes and they are rarely available currently for titration of catalytic antibody active sites. In the first part of the article we discuss a general kinetic method for the investigation of active-site availability in preparations of macromolecular catalysts. The method involves steady-state kinetics to provide Vmax and Km and single-turnover first-order kinetics using excess of catalyst over substrate to provide the analogous parameters k(obs)lim and K(m)app. The active-site contents of preparations that contain only active catalyst (Ea) and inert material (Ei) may be calculated as [Ea](T) = Vmax)/k(obs)lim. This is true even if nonproductive binding to E(a) occurs. For polyclonal catalytic antibody preparations, which may contain binding but noncatalytic material (Eb) in addition to Ea and Ei, the significance of Vmax/k(obs)lim is more complex but provides an upper limit to E(a). This can be refined by consideration of the relative values of Km and the equilibrium dissociation constant of EbS. Analysis of the Ea, Eb, Ei system requires the separate determination of Ei. For catalytic antibodies this may be achieved by analytical affinity chromatography using an immobilized hapten or hapten analog and an ELISA procedure to ensure the clean separation of Ei from the Ea + Eb mixture. PMID- 11384191 TI - Quantitative analysis of the time courses of enzyme-catalyzed reactions. AB - The catalytic properties of enzymes are usually evaluated by measuring and analyzing reaction rates. However, analyzing the complete time course can be advantageous because it contains additional information about the properties of the enzyme. Moreover, for systems that are not at steady state, the analysis of time courses is the preferred method. One of the major barriers to the wide application of time courses is that it may be computationally more difficult to extract information from these experiments. Here the basic approach to analyzing time courses is described, together with some examples of the essential computer code to implement these analyses. A general method that can be applied to both steady state and non-steady-state systems is recommended. PMID- 11384192 TI - The competition plot: A kinetic method to assess whether an enzyme that catalyzes multiple reactions does so at a unique site. AB - Enzymes often act on more than one substrate, and the question then arises as to whether this can be attributed to the existence of two different enzymes that have not been separated or, more interesting, to the presence of two different active sites in the same enzyme. The competition plot is a kinetic method that allows us to test with little experimentation whether the two reactions occur at the same site or at different sites. It consists of making mixtures of the two substrates and plotting the total rate against a parameter p that defines the concentrations of the two substrates in terms of reference concentrations chosen to give the same rates at p = 0 and p = 1, i.e., when only one of the substrates is present. With a slight modification of the equations it can also be applied to enzymes that deviate from Michaelis-Menten kinetics. If the two substrates react at the same site, the competition plot gives a horizontal straight line; i.e., the total rate is independent of p. In contrast, if the two reactions occur at two separate and independent sites a curve with a maximum is obtained; separate reactions with cross-inhibition generate curves with either maxima or minima according to whether the Michaelis constants of the two substrates are smaller or larger than their inhibition constants in the other reactions. Strategies to avoid ambiguous results and to improve the sensitivity of the plot are described. A practical example is given to facilitate the experimental protocol for this plot. PMID- 11384193 TI - Detection of errors of interpretation in experiments in enzyme kinetics. AB - Although modern statistical computing will often be the method of choice for analyzing kinetic data, graphic methods provide an important supplement that ought not to be neglected. Residual plots, or plots of differences between observed and calculated values against variables not expected to be correlated with these differences, permit a rapid judgment of whether data have been correctly interpreted and analyzed. The rapid increase in the frequency with which artificially modified or mutated enzymes are studied is making it less and less safe to assume that enzymes are stable under assay conditions, and there is thus an increased need for methods to check for enzyme stability, and a method for doing this is briefly described. Finally, the Scatchard plot (together with the Eadie-Hofstee plot) is used as an example to discuss the dangers of publishing derived information unaccompanied by any primary data. PMID- 11384194 TI - Peroxisomal deficiencies are associated with altered activity of endothelial NOS in human fibroblasts. AB - As shown recently, in human skin fibroblasts both a constitutively expressed and the inducible nitric oxide synthase (NOS) isoform are present. To identify the NOS isoforms expressed under standard conditions in healthy human skin fibroblasts and fibroblasts with peroxisomal deficiencies (cell lines from patients suffering from X-chromosome linked Adrenoleukodystrophy (X-ALD) and the Zellweger Syndrome), we cultivated the cells in Dulbecco's modified Eagle's medium without inflammatory mediators. Our experiments clearly showed that human fibroblasts with and without peroxisomal deficiencies only contain the constitutively expressed endothelial nitric oxide synthase (eNOS) isoform and that the eNOS is tyrosine-phosphorylated. The inducible isoform (iNOS) could not be detected under standard conditions. Healthy human skin fibroblasts show a higher specific NOS activity than X-ALD and Zellweger cells (2.25 to 1.68 and 1.17 pmol L-citrulline/min/mg total cellular protein), although the content of eNOS protein does not differ significantly in these cell lines. However the tyrosine-phosphorylated portion of eNOS is significantly lower in X-ALD and Zellweger cells. PMID- 11384195 TI - Nitric oxide in skeletal muscle: inhibition of nitric oxide synthase inhibits walking speed in rats. AB - Nitric oxide (NO*) is a multifunctional messenger molecule generated by a family of enzymes called the nitric oxide synthases (NOSs). Although NOSs have been identified in skeletal muscle, specifically brain NOS (bNOS) and endothelial NOS (eNOS), their role has not been well clarified. The goals of this investigation were to (1) characterize the immunoreactivity, Ca(2+) dependence, and activity of NOS in human and rat skeletal muscle and (2) using a rat model, investigate the effect of chronic blockade of NOS on skeletal muscle structure and function. Our results showed that both human and rodent skeletal muscle had NOS activity. This NOS activity was similar to that of the endothelial and brain NOS isoforms in that it was calcium-dependent. However, Western blot analysis consistently showed that a polyclonal antibody raised against a peptide sequence of human inducible NOS (iNOS) reacted with a protein with a molecular weight (95 kDa) that was different from that of other NOS isoforms. RT-PCR analysis identified the mRNA expression of not only eNOS and bNOS but also iNOS in human and rat muscle. Inhibition of nitric oxide synthase in rats with N(omega)-nitro-L-arginine methyl ester (L-NAME) resulted in a progressive, severe reduction in walking speed (30 fold reduction in walking velocity at day 22, P < 0.001), muscle fiber cross sectional area (40% reduction at day 22, P < 0.001), and muscle mass (40% reduction in dry weight at day 22, P < 0.01). Rats fed the same regimen of the enantiomer of L-NAME (d-NAME) had normal motor function, muscle fiber morphology, and muscle mass. Taken together, these results imply that there may be a novel nitric oxide synthase in muscle and that NO. generated from muscle may be important in muscle function. PMID- 11384196 TI - Synergism between nitric oxide and hydrogen peroxide in the inhibition of platelet function: the roles of soluble guanylyl cyclase and vasodilator stimulated phosphoprotein. AB - In previous studies, a strong synergism between low concentrations of hydrogen peroxide and nitric oxide in the inhibition of agonist-induced platelet aggregation has been established and may be due to enhanced formation of cyclic GMP. In this investigation, hydrogen peroxide and NO had no effect on the activity of pure soluble guanylyl cyclase or its activity in platelet lysates and cytosol. H(2)O(2) was found to increase the phosphorylation of vasodilator stimulated phosphoprotein (VASP), increasing the amount of the 50-kDa form that results from phosphorylation at serine(157). This occurs both in the presence and in the absence of low concentrations of NO, even at submicromolar concentrations of the peroxide, which alone was not inhibitory to platelets. These actions of H(2)O(2) were inhibited to a large extent by an inhibitor of cyclic AMP-dependent protein kinase, even though H(2)O(2) did not increase cyclic AMP. This inhibitor reversed the inhibition of platelets induced by combinations of NO and H(2)O(2) at low concentrations. The results suggest that the action on VASP may be one site of action of H(2)O(2) but that this event alone does not lead to inhibition of platelets; another unspecified action of NO is required to complete the events required for inhibition. PMID- 11384197 TI - Organ sites of lipopolysaccharide-induced nitric oxide production in the anesthetized rat. AB - The objective of this research was to determine the amount and timing of nitric oxide (NO, nitrogen monoxide) gas produced by the lungs, intestinal mucosa, and organ surfaces facing the peritoneal cavity after iv injection of a bacterial toxin, lipopolysaccharide (LPS). Some of the deleterious effects of LPS on organ function have been attributed to NO or strong oxidants formed locally from NO. Medical-grade air was used as an inspiratory air source (50 strokes/min x 3 ml/stroke) or was pumped through the ileal lumen or peritoneal cavity (20 strokes/min x 3 ml/stroke). The air was collected at intervals of 15-30 min for 3 h after LPS and analyzed for authentic NO gas by chemiluminescence. LPS (5 mg/kg) or saline was injected iv. Sodium nitroprusside (SNP) was injected to determine the appearance of its NO released into the perfused compartments. Blood pressure, plasma nitrate plus nitrite (NO(x)), and total plasma leukocytes were measured as other manifestations of LPS effects. NO began to increase in the pulmonary expired air 90 min after LPS and continued to increase for the remainder of the experiment. The final pulmonary post-LPS [NO] was about 20-fold greater than the [NO] before LPS. LPS had no effect on intraluminal or intraperitoneal [NO]. The saline injection had no effect on [NO] in any compartment. SNP injection increased NO entry into all three air-perfused compartments. Thus, NO from an exogenous tissue source was not prevented from being detected. Blood pressure was decreased by LPS only during the pulmonary perfusion. There were no significant effects of LPS on leukocytes or plasma NO(x). LPS decreased blood pressure and leukocytes and increased plasma NO(x) when air perfusion was not done. It was concluded that different organs can produce LPS-induced NO at markedly different rates and times. However, some aspect of the experimental technique of air perfusion could alter the effects of LPS. PMID- 11384198 TI - Coinduction of endothelial nitric oxide synthase and arginine recycling enzymes in aorta of diabetic rats. AB - Decreased availability of arginine and impaired production of NO (nitric oxide) have been implicated in the development of endothelial dysfunction. Citrulline formed by the NOS reaction is recycled to arginine by the citrulline-NO cycle, which is composed of NOS, argininosuccinate synthetase (AS), and argininosuccinate lyase. Therefore, we investigated the alterations of these enzymes in the aorta of streptozotocin (STZ)-induced diabetic rats. eNOS and AS mRNAs were increased by three- to fourfold 1-2 weeks after STZ treatment and decreased at 4 weeks. AL mRNA was weakly induced. Induction of eNOS and AS proteins was also observed. Cationic amino acid transporter (CAT)-1 mRNA remained little changed, and CAT-2 mRNA was not detected. The plasma nitrogen oxide levels were increased 1-2 weeks after STZ treatment and decreased at 4 weeks. Transforming growth factor-beta1 (TGF-beta1) mRNA in the aorta was also induced. TGF-beta1 induced eNOS and AS mRNAs in human umbilical vein endothelial cells but inhibited the proliferation of HUVEC. These results indicate that eNOS and AS are coinduced in the aorta in early stages of STZ-induced diabetic rats and that the induction is mediated by TGF-beta1. The results also suggest that TGF-beta1 works antiatherogenically at early stages of diabetes by increasing NO production, whereas prolonged elevation of TGF-beta1 functions atherogenically by inhibiting endothelial cell growth. PMID- 11384199 TI - Inhibitory effects of nitric oxide on oxidative phosphorylation in plant mitochondria. AB - Plant nitrate reductase (NR) produces nitric oxide (NO) when nitrite is provided as the substrate in the presence of NADH [H. Yamasaki and Y. Sakihama (2000) FEBS Lett. 468, 89-92]. Using a NR-dependent NO producing system, we investigated the effects of NO on the energy transduction system in plant mitochondria isolated from mung bean (Vigna radiata). Plant mitochondria are known to possess two respiratory electron transport pathways-the cytochrome and alternative pathways. When the alternative pathway was inhibited by n-propyl gallate, the addition of NR strongly suppressed respiratory O(2) consumption driven by the cytochrome pathway. In contrast, the alternative pathway measured in the presence of antimycin A was not affected by NO. The extent of the steady-state membrane potential (Deltapsi) generated by respiratory electron transport rapidly declined in response to NO production. The addition of bovine hemoglobin, a quencher of NO, resulted in the recovery of Deltapsi to the uninhibited level. Consistent with its inhibition of Deltapsi, NO produced by NR strongly suppressed ATP synthesis in the mitochondria. These results provide substantial evidence to confirm that the plant alternative pathway is resistant to NO and support the idea that the alternative pathway may lower respiration-dependent production of active oxygens under conditions where NO is overproduced. PMID- 11384200 TI - Nitric oxide regulation of lingual blood flow in the rat. AB - The purpose of this study was to determine the role of nitric oxide in the maintenance of basal lingual blood flow in the anesthetized rat. By using laser Doppler flowmetry, blood flow was measured from the tongue before and after treatment with the nonselective inhibitor of nitric oxide synthase, L-NAME (0.2, 2.0, and 20 mg/kg), or the selective neuronal nitric oxide synthase inhibitor, 7 nitroindazole (40 mg/kg). Other groups of rats were treated with saline, D-NAME (2.0 mg/kg), L-arginine (200 mg/kg), L-arginine + L-NAME (200 + 2.0 mg/kg), or the 7-nitroindazole vehicle. L-NAME produced a dose-related depression in blood flow in the tongue (concurrent with increased arterial blood pressure), which was attenuated by prior administration of L-arginine. Lingual blood flow depression was not seen after administration of the inactive stereoisomer, D-NAME. In addition, the neuronally specific nitric oxide synthase inhibitor, 7 nitroindazole, failed to produce a significant depression of lingual blood flow. These results suggest that the tonic release of nitric oxide from the vascular endothelium plays an important role in maintaining basal blood flow in the tongue and that neuronally released nitric oxide is not involved in maintaining basal circulation in this vascular bed. PMID- 11384201 TI - N-hydroxybenzenecarboximidic acid derivatives: a new class of nitroxyl-generating prodrugs. AB - On the basis of the propensity of Piloty's acid to generate nitroxyl (HNO), we previously prepared a number of N,O-bisacylated Piloty's acid derivatives and showed that such prodrugs underwent a disproportionation reaction following ester hydrolysis to give an unstable intermediate that hydrolyzed to nitroxyl. To expand the versatility of this series, we desired some mixed N,O-diacylated Piloty's acid derivatives and devised a synthetic route to them. Such efforts led us, serendipitously, to a new series of heretofore unreported nitroxyl-generating compounds. Thus, benzohydroxamic acid was acylated on the hydroxylamino oxygen and the resulting product converted to its sodium salt. Treatment of this salt with arenesulfonyl chorides would be expected to give the mixed N,O-diacylated derivatives of Piloty's acid. However, the products obtained were the isomeric carboximidic acid derivatives whose structures were deduced from the IR and (13)C NMR spectral frequencies associated with the sp(2) carbons. The structures were verified by analysis of the X-ray crystal structure of a prototype compound of this series. When incubated with porcine liver esterase or mouse plasma, these N acyloxy-O-arenesulfonylated benzenecarboximidic acid derivatives liberated HNO, measured as N(2)O, as well as the expected arenesulfinic acid and benzoic acid. Alkaline hydrolysis also produced N(2)O, but the major products were the arenesulfonic acid and benzohydroxamic acid. Thus, these N hydroxybenzenecarboximidic acid derivatives represent a new series of nitroxyl prodrugs that require enzymatic bioactivation before nitroxyl can be liberated. PMID- 11384202 TI - Repeated restraint stress-induced increase in baroreceptor reflex sensitivity: role of corticotropin-releasing factor. AB - The effect of central administration of a corticotropin-releasing factor antagonist on repeated restraint stress-induced changes in the baroreceptor reflex response to phenylephrine was examined in male Wistar-Kyoto rats. Rats were instrumented with intracerebroventricular guide cannula and femoral arterial and venous catheters. On each of 5 consecutive days, two groups received either a central infusion of saline or an infusion of the corticotropin-releasing factor antagonist, astressin. One saline- and one astressin-treated group experienced 20 min of restraint stress 10 min after each infusion. The other saline- and astressin-treated groups served as non-stressed controls. Twenty-four hours later, each rat received 3 doses of phenylephrine which produced equivalent increases in mean arterial pressure in each of the 4 treatment groups. Reflex bradycardia was significantly greater in the saline-treated/repeated restraint group than in the saline-treated/no restraint group. This effect of repeated restraint on the baroreceptor reflex was attenuated by administration of astressin prior to each session of restraint. A single 20 min session of restraint stress failed to alter baroreceptor reflex sensitivity. However, repeated central infusions of exogenous CRF failed to alter BRR sensitivity. In a separate experiment, astressin failed to attenuate the increases in mean arterial pressure and heart rate which occurred during each session of restraint stress and, in fact, diminished habituation of the blood pressure response in the last session. The results suggest that repeated stress increases thesensitivity of the baroreceptor reflex and that corticotropin-releasing factor has a role in this stress-induced change. PMID- 11384203 TI - Pharmacological studies on the monoaminergic influence on the synthesis and expression of neuropeptide Y and corticotropin releasing factor in rat brain amygdala. AB - Our earlier findings concerning the 6-OHDA lesion suggested dopaminergic regulation of neuropeptide Y (NPY) and corticotropin releasing factor (CRF) synthesis and expression in amygdala neurons. On the other hand, some other studies indicated that not only dopamine, but also other monoamines may modulate peptidergic neurons. Therefore the present study examined the effect of pharmacological deprivation of monoaminergic influences on NPY and CRF neurons in rat brain amygdala by means of in situ hybridization and immunohistochemical methods. It was found that NPY mRNA expression in the amygdala decreased after 24h blockade of dopaminergic D1 and D2 receptors, by haloperidol or SCH23390. At the same time the NPY-peptide expression measured immunohistochemically was not significantly changed. A prolonged, 14-day, blockade of dopaminergic receptors by haloperidol induced an opposite effect, an increase in NPY mRNA expression. Impairment of the serotonergic transmission by blockade of 5-HT synthesis using p chlorophenylalanine, as well as attenuation of the noradrenergic transmission by NA depletion from terminals by DSP4, did not significantly change NPY mRNA expression or the mean number of NPY-immunoreactive neurons in the amygdala. Only a decrease in the staining intensity observed as a decreased number of darkly stained neurons was found after both compounds. Neither the dopamine receptor blockade nor the impairment of serotonergic or noradrenergic transmission changed CRF mRNA or the peptide expression in the amygdala. The obtained results indicate that in rat brain amygdala, of all the monoamines, dopamine seems to be the most important modulator of NPY biosynthesis and expression. The effect of blockade of dopaminergic receptors is biphasic: first it induces a decrease and then - after prolonged treatment an increase in NPY mRNA. Serotonergic and noradrenergic systems in the amygdala seem to be connected with regulation of NPY release rather than the biosynthesis. PMID- 11384204 TI - Tachykinin activation of human monocytes from patients with rheumatoid arthritis: in vitro and ex-vivo effects of cyclosporin A. AB - Three types of tachykinin receptors, namely NK1, NK2 and NK3, are known to preferentially interact with substance P (SP), neurokinin A (NKA) and neurokinin B (NKB), respectively. We previously demonstrated that NK1 and NK2 receptors are present on human monocytes, SP and NKA inducing superoxide anion production and tumor necrosis factor-alpha (TNF-alpha) mRNA expression. NK2 receptor stimulation also triggered an enhanced respiratory burst in monocytes isolated from rheumatoid arthritis (RA) patients. This study was aimed to evaluate the in vitro and ex-vivo effects of cyclosporin A (CsA) on tachykinins-evoked TNF-alpha release from monocytes of healthy donors and RA patients. CsA (100 ng/ml) potently inhibited phorbol ester- and tachykinin-evoked TNF-alpha secretion. In RA patients treated with CsA (Sandimmun Neoral 2.5 mg/kg/day, a significant time dependent reduction in TNF-alpha secretion from monocytes was measured. This may contribute to the CsA therapeutic activity in RA. PMID- 11384205 TI - The effects of CRA 1000, a non-peptide antagonist of corticotropin-releasing factor receptor type 1, on adaptive behaviour in the rat. AB - Intracerebrally administered CRF has been demonstrated to elicit several behavioural deficits in novel and potentially stressful experimental paradigms, and to promote activity in familiar situations. This study examined the effect of CRA 1000, a novel non-peptide antagonist of CRF(1)receptors, on rat behaviour in tests of anxiolytic and antidepressant activity and novelty-oriented behaviour. CRA 1000 (1.25-10 mg/kg) had no major effect in elevated plus-maze and social interaction tests. However, CRA 1000 (5 mg/kg) significantly reduced immobility in the forced swimming test, suggesting an antidepressant-like effect. In the exploration box test, CRA 1000 (1.25 mg/kg) had an anxiolytic effect on rat exploratory behaviour both in intact rats and after lesioning of the projections of locus coeruleus by DSP-4 (50 mg/kg) treatment. A higher dose of CRA 1000 (5 mg/kg) tended to have anxiolytic-like effects in DSP-4 pretreated rats, but in intact animals this dose prevented the increase in exploration which develops with repeated exposure to initially anxiety-provoking situations. Taken together, these experiments demonstrate that CRF1 receptor blockade by CRA 1000 has antidepressant-like effects, does not have a robust anti-anxiety effect in non stressed animals, but does have anxiolytic-like effects in more complex tasks, which can be observed also after denervation of the locus coeruleus projections. However, large doses of CRF1 receptor antagonists may reduce motivation of exploratory behaviour in familiar environments. PMID- 11384206 TI - The effect of paracetamol on nociception and dynorphin A levels in the rat brain. AB - Male Wistar rats were administered with naloxone (1 mg/kg i.p.) or MR 2266 (5 mg/kg i.p) 15 min before paracetamol (400 mg/kg i.p.) treatment and the pain threshold was evaluated. Rats were subjected to the hot-plate and formalin tests and immunoreactive dynorphin A (ir-dynorphin A) levels were measured in the hypothalamus, hippocampus, striatum, brainstem, frontal and parietal-temporal cortex by radioimmunoassay. Pretreatment with naloxone abolished paracetamol antinociceptive activity both in hot-plate and in the first phase, but not in the second phase of the formalin test, while MR 2266 pretreatment was able to antagonise paracetamol effect either in the hot-plate test or in both phases of the formalin test. Among different brain areas investigated paracetamol significantly decreased ir-dynorphin A levels only in the frontal cortex. MR 2266 but not naloxone reversed the decrease in ir-dynorphin A levels elicited by paracetamol. Paracetamol seems to exert its antinociceptive effect also through the opioidergic system modulating dynorphin release in the central nervous system (CNS) of the rat, as suggested by the decrease in the peptide levels. PMID- 11384207 TI - The polypeptide PHI discriminates a GTP-insensitive form of VIP receptor in liver membranes. AB - In early reports on 125I-VIP binding experiments in liver membranes, it has been proposed that, the VIP binding sites were partially sensitive to GTP. Here we confirm that the VIP binding sites of chicken liver membranes consisted mainly in bivalent VIP/PACAP receptors and that about 50% of the 125I-VIP binding capacity was not affected by the GTP analogue GppNHp. Part of these bivalent receptors also appeared to represent PHI binding sites. In GppNHp-treated membranes, the GTP-insensitive VIP binding sites displayed a 17-fold higher relative affinity than in control membranes for the VIP analogue PHI. Such data suggested that GTP insensitive VIP receptors may correspond to a subclass of high-affinity PHI receptors. Cross-linking of 125 I-VIP or 125 I-PHI to their receptors, revealed 2 components of 48 and 60 kDa. The radiolabelling of the 60 kDa component was strongly affected by increasing concentrations of the GTP analogue but was modestly abolished by an excess of PHI. Conversely, the radiolabelling of the 48 kDa molecular form was not affected by the GTP analogue but was efficiently abolished by increasing concentrations of PHI. Taken together, the data suggest that the 48 kDa component expressed in chicken liver membranes display the properties of a GTP-insensitive VIP/PHI receptor that can be pharmacologically discriminated from the GTP-sensitive 60 kDa form, through its much higher affinity for PHI. PMID- 11384208 TI - Acute cocaine alters oxytocin levels in the medial preoptic area and amygdala in lactating rat dams: implications for cocaine-induced changes in maternal behavior and maternal aggression. AB - Acute cocaine administration has been correlated with disruptions in the onset and maintenance of maternal behavior as well as decreases in maternal aggressive behavior in rat dams. A growing body of evidence suggests that cocaine may alter oxytocin levels leading to impairments in maternal behavior and aggression. The current study assessed whether acute cocaine injections alter oxytocin (OT) levels in the medial preoptic area (MPOA), ventral tegmental area (VTA), amygdala (AMY), and hippocampus (HIP) on postpartum day (PPD) 1 or PPD 6. On PPD 1, 30 mg/kg cocaine reduced OT levels by approximately 26.9% (picograms/milligram) in the MPOA (t (18) = 3.44, P<.01) compared to saline. On PPD 6, 30 mg/kg cocaine significantly increased OT levels by approximately 20.9% (picograms/brain area) in the AMY (F (2,25) = 3.44, P=.05) relative to saline. These findings suggest that acute cocaine may disrupt maternal behavior and maternal aggression at least in part through its action on the oxytocinergic system. PMID- 11384209 TI - Disposition of polychlorinated dibenzo-p-dioxins, dibenzofurans, and non-ortho polychlorinated biphenyls in pregnant long evans rats and the transfer to offspring. AB - Pharmacokinetic properties of polychlorinated dibenzo-p-dioxins, dibenzofurans (PCDFs), and non-ortho biphenyls (PCBs) play a critical role in their relative toxicity. The present study examined the transfer of these chemicals to offspring and placenta. Pregnant Long Evans rats received 0.0 (control), 0.05, 0.2, 0.8, and 1.0 microg/kg of dioxin toxic equivalence (TEQ) by oral gavage on the 15th gestational day (GD 15), using a dosing mixture that contained 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD), 2,3,7,8-tetrachlorodibenzofuran (TCDF), 1,2,3,7,8-pentachlorodibenzo-p-dioxin (PeCDD), 1,2,3,7,8-pentachlorodibenzofuran (1-PeCDF), 2,3,4,7,8-pentachlorodibenzofuran (4-PeCDF), octachlorodibenzofuran (OCDF), 3,3',4,4'-tetrachlorobiphenyl (PCB 77), 3,3',4,4',5-pentachlorobiphenyl (PCB 126), and 3,3',4,4',5,5'-hexachlorobiphenyl (PCB 169) in ratios approximating that in food. Rats were euthanized on GD 16, GD 21, and postnatal day 4 (PND 4). The chemical concentrations in fetus, pup, placenta, and maternal liver, serum, and adipose tissue were determined using gas chromatography/high resolution mass spectrometry. A dose-dependent increase in hepatic sequestration was seen with TCDD, PeCDD, 4-PeCDF, OCDF, PCB 126, and PCB 169, and the transfer to offspring was reduced at higher doses. 4-PeCDF, PeCDD and PCB 126 showed higher liver affinity than TCDD. TCDF, 1-PeCDF, and PCB 77 were metabolized rapidly. On GD 16, TCDD and the three PCBs reached equilibration between the fetus and placenta, but this did not occur with PeCDD and 4-PeCDF until GD 21, according to the lipid-based concentrations. Offspring compartments received more of the dosed compounds lactationally than transplacentally (7-28% versus 0.5-3%). The behavior of each congener was dose-dependent; therefore, extrapolation of high-dose experimental data should be used with caution. PMID- 11384210 TI - Protection of the Cyp1a2(-/-) null mouse against uroporphyria and hepatic injury following exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin. AB - The effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on the liver of C57BL/6J mice is a model for clinical sporadic porphyria cutanea tarda (PCT). There is massive uroporphyria, inhibition of uroporphyrinogen decarboxylase (UROD) activity, and hepatocellular damage. A variety of evidence implicates the CYP1A2 enzyme as necessary for mouse uroporphyria. Here we report that, 5 weeks after a single oral dose of TCDD (75 microg/kg), Cyp1a2(+/+) wild-type mice showed severe uroporphyria and greater than 90% decreases in UROD activity; in contrast, despite exposure to this potent agent Cyp1a2(-/-) knockout mice displayed absolutely no increases in hepatic porphyrin levels, even after prior iron overload, and no detectable inhibition of UROD activity. Plasma levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST)-although elevated in both genotypes after TCDD exposure-were significantly less in Cyp1a2(-/-) than in Cyp1a2(+/+) mice, suggesting that the absence of CYP1A2 also affords partial protection against TCDD-induced liver toxicity. Histological examination confirmed a decrease in hepatocellular damage in TCDD-treated Cyp1a2(-/-) mice; in particular, there was no bile duct damage or proliferation that in the Cyp1a2(+/+) mice might be caused by uroporphyrin. We conclude that CYP1A2 is both necessary and essential for the potent uroporphyrinogenic effects of TCDD in mice, and that CYP1A2 also plays a role in contributing to TCDD-induced hepatocellular injury. This study has implications for both the toxicity assessment of TCDD and the hepatic injury seen in PCT patients. PMID- 11384211 TI - 1,25-Dihydroxyvitamin D(3) prevents the conversion of adipose tissue into fibrous tissue in skin exposed to chronic UV irradiation. AB - The hormonally active form of vitamin D (1,25(OH)2D3) is known to be a physiological regulator of the proliferation and differentiation of skin cells including keratinocytes, fibroblasts, and adipocytes. In the present study, the efficacy of 1,25(OH)2D3 on the conversion of adipose tissue to fibrous tissue in photodamaged skin was investigated in a murine animal model. Groups of hairless mice were exposed to solar-simulating UV irradiation (lambdamax, 352 nm; UV distribution: 300-310 nm, 0.9%; 310-320 nm, 2.0%; 320-420 nm, 97.1%) for 20 weeks at a dose of 10.8 J/cm2 five times weekly on weekdays. At the end of 20 weeks irradiation, wrinkling in the dorsal skin was induced. The histological and biochemical studies indicated that UV irradiation caused a disappearance of adipocytes and concomitant accumulation of the extracellular matrix components (fibrosis), including collagen, hyaluronan, and chondroitin/dermatan, which are synthesized by fibroblasts. Application of 1,25(OH)2D3 on the dorsal skin prior to UV irradiation dramatically prevented both the disappearance of adipocytes and the accumulation of extracellular matrix components in the lower dermis, resulting in antiwrinkling. These findings indicate that 1,25(OH)2D3 prevents the UV-induced abnormal differentiation and proliferation of adipocytes and fibroblasts, which arise from a common progenitor, mesenchymal cells. PMID- 11384212 TI - Hydrocarbon carcinogens evade cellular defense mechanism of G1 arrest in nontransformed and malignant lung cell lines. AB - In previous studies using human breast carcinoma cells (MCF-7) and human colon carcinoma cells (RKO) we have shown that, in response to treatment with hydrocarbon carcinogens, these cell lines failed to undergo a p53-mediated cell cycle arrest in G1 phase; rather, the cells were accumulated in the S phase with damaged DNA, a situation that may lead to replication of DNA on a damaged template, resulting in the enhanced frequency of mutations in the daughter cells. This has been termed a stealth effect. In the present work we have demonstrated that the stealth effect also pertains for lung cells. In E10 nontransformed mouse lung type II cells, two potent hydrocarbon carcinogens, benzo[a]pyrene dihydrodiol epoxide and benzo[g]chrysene dihydrodiol epoxide, damaged DNA as suggested by retardation in S phase, but did not cause G1 arrest, in contrast to the positive control, actinomycin D. Human lung adenocarcinoma A549 cells, with normal p53, likewise exhibited G1 arrest after actinomycin D, but not after treatment with the diol epoxides. Several human lung cancer cell lines with absent or mutant p53, such as H358, H1734, and H82, exhibited no G1 arrest after any of the compounds. However, lung H441 adenocarcinoma cells, with a mutation in exon 5, codon 158 of p53, exhibited partial G1 arrest after the diol epoxides as well as actinomycin D, and H2030 adenocarcinoma cells did not show G1 arrest after any of the chemicals despite a normal p53. The stealth effect of evasion of G1 arrest may contribute to initiation of lung adenocarcinomas and to progression of tumors. A role in resistance to chemotherapy by certain drugs is also likely. PMID- 11384213 TI - Inhaled naphthalene causes dose dependent Clara cell cytotoxicity in mice but not in rats. AB - Current OSHA standards for naphthalene exposure are set at 10 ppm (time-weighted average) with a standard threshold exposure concentration of 15 ppm. While several studies have thoroughly delineated the time course and dose response of injury by naphthalene administered ip, the pattern and severity of injury by inhalation exposure are unknown. These studies compare the regiospecific and dose dependent cytotoxicity of naphthalene after inhalation exposure. Mice and rats were exposed for 4 h to naphthalene vapor at concentrations of 0-110 ppm. In rats, no injury was observed in the lung epithelium at exposure concentrations up to 100 ppm. Exposures as low as 2 ppm produced proximal airway injury in mice, with increased severity in a concentration-dependent fashion up to 75 ppm. Terminal airways of exposed mice exhibited little or no injury at low concentrations (1-3 ppm). Exposures of 8.5 ppm or higher were required to produce injury to Clara cells in the terminal airways. In contrast, administration of naphthalene (300 mg/kg) extended the injury pattern toward the lobar bronchus. We conclude (1) the pattern of injury to naphthalene is highly dependent on the route of exposure, (2) lung injury to inhaled naphthalene is species dependent, and (3) Clara cells of mouse airways are exquisitely sensitive to inhaled naphthalene at concentrations well below the current OSHA standard for human exposure. PMID- 11384214 TI - In vivo canine model comparison of cardiohemodynamic and electrophysiological effects of a new antipsychotic drug aripiprazole (OPC-14597) to haloperidol. AB - The cardiovascular effects of aripiprazole were assessed in comparison with those of haloperidol using a halothane-anesthetized canine model with monophasic action potential monitoring. Aripiprazole (n = 6) or haloperidol (n = 6) was infused over 10 min at escalating doses of 0.03, 0.3, and 3.0 mg/kg with intervals of 20 min between doses. Clinically relevant plasma concentrations were obtained after 0.03-0.3 mg/kg of aripiprazole as well as haloperidol. After 0.03-0.3 mg/kg of aripiprazole, positive chronotropic, inotropic, and dromotropic effects, shortening of the ventricular effective refractory period (ERP) and repolarization phase, and decrease of total peripheral resistance were observed in a dose-related manner. However, in the presence of a beta-blocking dose of esmolol (0.1 mg/kg/min), these changes were not induced. After 3.0 mg/kg of aripiprazole administration, cardiac effects induced by the lower doses were attenuated or disappeared, while the negative chronotropic, dromotropic, and hypotensive actions and prolongation of ERP and repolarization phase were induced. After 0.03 mg/kg of haloperidol, no significant change was observed, except for the decrease of the peripheral resistance. After 0.3-3.0 mg/kg of haloperidol, negative chronotropic, inotropic, and hypotensive actions, intraventricular conduction delay, and prolongation of ventricular ERP and repolarization phase were observed in a dose-related manner accompanied by further decrease of the peripheral resistance. The inhibitory effects of aripiprazole on cardiovascular parameters in dogs were less potent than those of haloperidol at clinically relevant exposures, moreover, aripiprazole, unlike haloperidol, neither induced early afterdepolarization nor prolonged the ventricular electrical vulnerable period. Therefore, aripiprazole can be considered safer to use than haloperidol. PMID- 11384215 TI - Varicella vaccine: genesis, efficacy, and attenuation. PMID- 11384216 TI - Role of c-myc regulation in Zta-mediated induction of the cyclin-dependent kinase inhibitors p21 and p27 and cell growth arrest. AB - Latency-associated Epstein-Barr virus (EBV) gene expression induces cell proliferation. Unlike the latency associated genes, lytic gene expression in EBV, as well as other herpesviruses, elicits cell cycle arrest. Previous studies have shown that the EBV immediate early lytic transactivator, Zta, induces a G(0)/G(1) cell cycle arrest through induction of the cyclin-dependent kinase inhibitors, p21 and p27. Here we show that while EBV latency is intimately linked to activation of the protooncogene, c-myc, Zta represses c-myc expression. We also show that inhibition of c-myc expression is required for Zta-mediated growth arrest and for maximal induction of p21 and p27. Nevertheless, induction of p21 and p27 is also influenced by a c-myc-independent mechanism. A detailed genetic analysis of Zta's basic/DNA binding region identified two distinct subregions that contribute to full induction of p21 and p27. One subdomain influences p21 and p27 expression through the c-myc-dependent mechanism and the other subdomain influences p21 and p27 induction through the c-myc-independent pathway. Together, these studies further our understanding of the complex nature of Zta-induced growth arrest. PMID- 11384217 TI - Identification of a novel occlusion derived virus-specific protein in Spodoptera exigua multicapsid nucleopolyhedrovirus. AB - Understanding the molecular basis of the distinct biological properties of Spodoptera exigua multicapsid nucleopolyhedrovirus (SeMNPV), such as its narrow host range and high virulence, requires detailed information on the temporal expression and subcellular localization of SeMNPV gene products. The expression of two unique SeMNPV ORFs, 116 (Se116) and 117 (Se117), which show 45% amino acid similarity, was analyzed. Se116 and Se117 were expressed both in cultured cells and in larvae of S. exigua as polyadenylated transcripts of 0.80 and 0.75 kb, respectively. These transcripts initiated from ATCA(G/T)T promoter motifs, commonly found for baculovirus early genes. Se116 transcripts were detected with increasing abundance from 8 to 48 h p.i., whereas Se117 transcripts were present from 4 h p.i. and most abundantly at 24 h p.i. Western blot analysis of infected Se301 cells revealed 27- and 23-kDa proteins for Se116 and Se117, respectively. C terminal GFP-fusion proteins of Se116 and Se117 were primarily localized in the nucleus of Se301 cells. When Se301 cells were infected with SeMNPV, both GFP fusion proteins were localized in the virogenic stroma of the nucleus. While the function of the Se116 protein is still enigmatic, the Se117 protein appeared to be a structural protein associated with nucleocapsids of occlusion-derived SeMNPV virions but not of budded virus. PMID- 11384218 TI - Assessment of recombinants that arise from the use of a TMV-based transient expression vector. AB - A potential use of virus-based transient expression vectors is the large-scale production of commercial specialty products, which would require the inoculation of many acres of plants with the viral vector. However, there are several concerns about the widespread use of virus-based vectors. Among these are the spread of the engineered virus to susceptible plants and the generation and persistence of recombinant viruses in the environment. Using a Tobacco mosaic virus (TMV)-based transient gene expression vector, 30B, which expresses the jellyfish green fluorescent protein (30B-GFP), we describe the predominant types of hybrid viruses that developed in plants. In general, the recombinants deleted the foreign gene and repeated sequences, retaining only those sequences required for optimal replication and movement. In pathogenicity studies and challenge experiments designed to make a comparative assessment of the competitiveness of the recombinants with the parent virus, the recombinants had reduced vigor and were less competitive and pathogenic than TMV, a virus which is already present in the areas where tobacco is grown. PMID- 11384219 TI - Immunopathogenesis and immune modulation of Venezuelan equine encephalitis virus induced disease in the mouse. AB - The course of Venezuelan equine encephalitis (VEE) disease in immunodeficient and immunologically normal mice was compared to define the role of the immune system in this disease process. Immunocompetent mice infected with VEE exhibited a biphasic illness characterized by an early self-limiting lymphoid phase and a fatal CNS phase. The lymphoid phase of the illness was characterized by extensive viral replication within spleen, thymus, Peyer's patches, and lymph nodes, was accompanied by a high-titered serum viremia, and resolved with the production of VEE-specific IgM class antibody at 72 h postinfection (p.i.). Immunocompetent animals survived an average of 6.8 +/- 1.2 days before succumbing to fulminant encephalitis. In contrast, SCID mice infected with VEE showed a persistent replication of virus throughout all organs tested beginning at 24 h p.i. VEE infected SCID mice exhibited a severe spongiform encephalopathy with 100% mortality and an average survival time of 8.9 +/- 0.9 days. These studies indicated that the characteristic organ tropism of VEE in the mouse is due in large part to an early anti-viral state, the establishment of which is dependent upon the presence of an intact immune system. Finally, the CNS pathology in a VEE infected mouse had a significant immunologic component. However, in contrast to other neurovirulent alphaviruses, VEE was directly cytopathic for the cells of the CNS, even in the absence of an immune response. PMID- 11384220 TI - First outbreak of callitrichid hepatitis in Germany: genetic characterization of the causative lymphocytic choriomeningitis virus strains. AB - Callitrichid hepatitis (CH) is a highly fatal, rodent-borne zoonosis of New World primates (family Callitrichidae) caused by lymphocytic choriomeningitis virus (LCMV). It is unclear whether virulence in Callitrichidae is associated with specific genetic or phylogenetic markers of the virus as only a partial S RNA sequence of a single CH-associated isolate is known. In a period of 10 months, three pygmy marmosets (Cebuella pygmaea) and one Goeldi's monkey (Callimico goeldii) died from CH in a German zoo. LCMV was most likely transmitted by wild mice. Infection was associated with characteristic histopathological lesions in liver, brain, and lymphoid tissue. Virus sequences from all callitrichids and a captured mouse were > or =99.2% identical. LCMV strains from a pygmy marmoset and the Goeldi's monkey were isolated in cell culture and the 3.4-kb S RNA was completely sequenced. Both strains differed considerably in their genetic and phylogenetic characteristics from known LCMV strains, including the previously described CH-associated strain. These data show that CH is widespread and can be caused by distantly related LCMV strains. PMID- 11384221 TI - Genetic characterization and geographic distribution of rabies virus isolates in Brazil: identification of two reservoirs, dogs and vampire bats. AB - We analyzed 50 rabies virus samples isolated in Brazil from 12 dogs, 11 cats, 5 vampire bats, 15 cattle, 2 horses, 1 pig, 1 sheep, and 3 humans to investigate the molecular epidemiology of rabies viruses. We sequenced 203 nucleotides on the nucleoprotein gene by direct sequencing of the PCR-amplified products. All the isolates belonged to the genotype 1 and homology of the 203 nucleotides was at least 83.7% among isolates. The main reservoirs were estimated based on the homology of nucleotide sequences. Brazilian rabies virus isolates were clustered into two reservoir groups: dogs and vampire bats. All the dog-related rabies virus isolates showed nucleotide homology greater than 99.0%. Vampire bat-related rabies virus isolates showed nucleotide homology greater than 96.6% and could be further divided into subgroups corresponding to areas where viruses were isolated. These data suggest that circulating rabies variants belong to at least two different genotype clusters in Brazil and that these two clusters are maintained independently among vampire bats and dogs. PMID- 11384222 TI - Structural transitions of satellite tobacco mosaic virus particles. AB - Satellite tobacco mosaic virus (STMV) can undergo at least two physical transitions that significantly alter its mechanical and structural characteristics. At high pH the 17-nm STMV particles expand radially by about 5 A to yield particles having diameters of about 18 nm. This pH-induced transition is further promoted by aging of the virions and degradation of the RNA, so that swollen particles ultimately appear even at neutral pH. While the native 17-nm particles crystallize as orthorhombic or monoclinic crystals which diffract to high resolution (1.8 A), the enlarged 18-nm particles crystallize in a cubic form which diffracts to no better than 5 A. In the transition, not only do the capsid protein subunits move radially outward, but the helical RNA segments with which they interact do as well. This is noteworthy because it demonstrates that the RNA and the protein shell are capable of coordinated movement, and that neither structure is rigidly defined or independent of the other. Using atomic force microscopy, it can be shown that STMV particles, upon drying, lose their mechanical rigidity and undergo deformation. Virions initially 17 nm in diameter shrink to more uniform final sizes than do 18 nm, initially swollen particles. This transition appears to be irreversible, as the particles do not reassume their former size nor structural rigidity upon rehydration. Evidence is also presented that preparations of native virus and their crystals are naturally somewhat heterogeneous and contain a variety of particles of anomalous size. PMID- 11384223 TI - Human herpesvirus 8 envelope-associated glycoprotein B interacts with heparan sulfate-like moieties. AB - Cell-surface heparan sulfate (HS) serves as an initial attachment receptor for several herpesviruses. The gamma2-human herpesvirus-8 (HHV-8) or Kaposi's sarcoma associated herpesvirus DNA and transcripts have been detected in B cells, endothelial cells, macrophages, and epithelial cells. HHV-8 infects a variety of human and animal cell lines leading to latent or abortive infection. Our studies showed that this broad cellular tropism may be in part due to HHV-8's interaction with the ubiquitous host cell-surface HS-like molecules. HHV-8 binding to the target cells and the infection were inhibited by soluble heparan, a glycosaminoglycan (GAG) closely related to HS. Since HHV-8 gB possess a putative heparan-binding domain (HBD) in the extracellular domain, the interaction of gB with HS-like moieties was examined. Unlike gB of gamma1-Epstein-Barr virus and gamma2-murine herpesvirus 68, HHV-8 gB was expressed on the surface of the infected cell membranes and virion envelopes. Envelope-associated gB was made up of 75 and 54 kDa polypeptides forming disulfide-linked heterodimers and multimers. Rabbit anti-gB antibodies neutralized HHV-8 infection. Virion envelope associated gB specifically bound to heparan-agarose, which was eluted by high concentration of soluble heparan, but not by chondroitin sulfates. In vitro transcribed and translated products of gB gene specifically bound to heparan agarose beads, which was blocked by HS and heparan, but not by other GAGs such as chondroitin sulfates (A, B, and C), N-acetyl heparan, and de-N-sulfated heparan. Biotinylated gB peptide corresponding to the putative HBD also bound to heparan. These results suggest that gB plays an important role in the infectious process of HHV-8 and virus interaction with cell-surface HS-like moieties could be in part mediated by the envelope-associated gB. PMID- 11384224 TI - Evidence for positive selection driving the evolution of HIV-1 env under potent antiviral therapy. AB - In HIV-infected individuals treated with potent antiretroviral therapy, viable virus can be isolated from latently infected cells several years into therapy, due to the long life of these cells, ongoing replication replenishing this population, or both. We have analysed the V3 region of the HIV-1 env gene isolated from six patients who have undergone 2 years of potent antiretroviral therapy without frank failure of viral suppression. We show that in two (and possibly three) patients, the sequence changes between baseline virus and virus isolated from infected cells persisting 2 years into infection result from positive selection driving adaptive evolution, occurring either prior to or during therapy. Our analyses suggest low-level replication despite absence of drug resistance due to drug sanctuary sites, or to low-level ongoing replication in the presence of alterations in the selective environment during therapy, perhaps due to a decline in HIV-specific immune responsiveness or changes in target cell pools. In one patient, genetic divergence between baseline plasma and infected cells isolated during therapy may reflect the long half-life of some of these persistent cell populations and the divergence of viral subpopulations that occurred prior to therapy. PMID- 11384225 TI - Recombinant equine arteritis virus as an expression vector. AB - Equine arteritis virus (EAV) is the prototypic member of the family Arteriviridae, which together with the Corona- and Toroviridae constitutes the order Nidovirales. A common trait of these positive-stranded RNA viruses is the 3'-coterminal nested set of six to eight leader-containing subgenomic mRNAs which are generated by a discontinuous transcription mechanism and from which the viral open reading frames downstream of the polymerase gene are expressed. In this study, we investigated whether the unique gene expression strategy of the Nidovirales could be utilized to convert them into viral expression vectors by introduction of an additional transcription unit into the EAV genome directing the synthesis of an extra subgenomic mRNA. To this end, an expression cassette consisting of the gene for a green fluorescent protein (GFP) flanked at its 3' end by EAV-specific transcription-regulating sequences was constructed. This genetic module was inserted into the recently obtained mutant infectious EAV cDNA clone pBRNX1.38-5/6 (A. A. F. de Vries, et al., 2000, Virology 270, 84-97) between the genes for the M and the G(L) proteins. Confocal fluorescence microscopy of BHK-21 cells electroporated with capped RNA transcripts derived from the resulting plasmid (pBRNX1.38-5/6-GFP) demonstrated that the GFP gene was expressed in the transfected cells, while the gradual spread of the infection through the cell monolayer showed that the recombinant virus was replication competent. The development of the cytopathic effect was, however, much slower than in cells that had received equivalent amounts of pBRNX1.38-5/6 RNA, indicating that the vector virus had a clear growth disadvantage compared to its direct precursor. Immunoprecipitation analyses of proteins from metabolically labeled BHK-21 cells infected with supernatant of the transfected cultures confirmed that the recombinant virus vector was viable and expressed viral genes as well as the GFP gene. Reverse transcription-PCR of the viral mRNAs extracted from cells infected with the vector virus revealed that it directed the synthesis of nine instead of eight different EAV RNAs. These findings were corroborated by hybridization analyses. Mapping of the leader-to-body junctions of the ninth mRNA indicated that the 3' part of the GFP gene contains cryptic transcription signals which gave rise to at least five different RNA species ranging in size from 1277 to 1439 nt [without oligo(A) tract]. Furthermore, translation of the unintended mRNA resulted in the production of an extended version of the EAV M protein. Serial passage of the recombinant virus vector led to its gradual replacement by viral mutants carrying deletions in the GFP gene. The reduction in viral fitness associated with the insertion of the expression cassette into the EAV genome apparently caused genetic instability of the recombinant virus. PMID- 11384226 TI - Allpahuayo virus: a newly recognized arenavirus (arenaviridae) from arboreal rice rats (oecomys bicolor and oecomys paricola) in northeastern peru. AB - Allpahuayo virus was initially isolated from arboreal rice rats (Oecomys bicolor and Oecomys paricola) collected during 1997 at the Allpahuayo Biological Station in northeastern Peru. Serological and genetic studies identified the virus as a new member of the Tacaribe complex of the genus Arenavirus. The small (S) segment of the Allpahuayo virus prototype strain CLHP-2098 (Accession No. AY012686) was sequenced, as well as that of sympatric isolate CLHP-2472 (Accession No. AY012687), from the same rodent species. The S segment was 3382 bases in length and phylogenetic analysis indicated that Allpahuayo is a sister virus to Pichinde in clade A. Two ambisense, nonoverlapping reading frames were identified, which result in two predicted gene products, a glycoprotein precursor (GPC) and a nucleocapsid protein (NP). A predicted stable single hairpin secondary structure was identified in the intergenic region between GPC and NP. Details of the genetic organization of Allpahuayo virus are discussed. PMID- 11384227 TI - Basic amino acid residues in the V3 loop of simian immunodeficiency virus envelope alter viral coreceptor tropism and infectivity but do not allow efficient utilization of CXCR4 as entry cofactor. AB - In contrast to human immunodeficiency viruses type 1 and type 2 (HIV-1 and HIV-2, respectively), simian immunodeficiency virus (SIVmac) rarely uses CXCR4 (X4) for efficient entry into target cells. Basic amino acid residues in the V3 loop of HIV Env allow efficient coreceptor utilization of X4. Therefore, we investigated if similar changes in the SIVmac Env protein also mediate a coreceptor switch from CCR5 (R5) to X4. Functional analysis revealed that none of eight SIVmac variants, containing V3 regions with an overall charge between +4 and +10, efficiently utilized X4 as entry cofactor. Nonetheless, these alterations had differential effects on SIV coreceptor tropism and on Env expression levels. A single amino acid substitution of L328R, located near the tip of the V3 loop, resulted in grossly reduced Env expression levels and impaired viral infectivity. Notably, additional basic residues restored efficient Env expression and virion incorporation but not infectivity. In comparison to the L328R mutation, changes of P334K and D337K had little disruptive effects on SIVmac entry and replication. Interestingly, mutation of L320K and P321R disrupted coreceptor usage of GPR15 but not R5. These changes also impaired SIVmac replication in peripheral blood mononuclear cells (PBMC) derived from a Delta32/Delta32 donor but not in R5 expressing human or simian PBMC. Our results show that positively charged amino acid residues in the V3 loop affect SIVmac coreceptor tropism and infectivity but do not allow efficient utilization of X4. PMID- 11384228 TI - The use of recombinant baculoviruses for sustained expression of human cytomegalovirus immediate early proteins in fibroblasts. AB - The isolation of viruses with mutations in essential genes requires that they be propagated in cells expressing the wild-type proteins. This has been a particularly challenging problem for studying mutations in the human cytomegalovirus (HCMV) immediate early (IE) gene, IE2 86. In the past, we tried a number of approaches to derive human fibroblasts expressing wild-type IE2 86, but were unable to maintain expression of a fully functional protein. To overcome this obstacle, we developed a strategy whereby recombinant baculoviruses were used as vectors for the expression of HCMV IE proteins in primary human fibroblasts (FFs). The IE2 86 and IE1 72 cDNAs, as well as the genomic fragment of the UL122-123 region under the control of a chicken actin promoter, were introduced into the baculovirus genome by site-specific transposition in Escherichia coli. Recombinant "bacmid" DNAs were then transfected into Sf9 cells to generate recombinant baculoviruses. FFs infected at high m.o.i. with these baculoviruses expressed high levels of the HCMV protein for at least 1 week, as determined by immunofluorescence assays and Western blots. Moreover, the IE2 86 protein was found to be fully functional with respect to its ability to activate the HCMV UL112-113 early promoter. Recombinant baculoviruses expressing IE1 72 were also able to efficiently complement HCMV ie1 mutants. These data demonstrate the potential of using recombinant baculoviruses as vectors for the expression of toxic viral genes in human cells and for subsequent isolation of mutant HCMV lacking these essential genes. PMID- 11384229 TI - Mutations in the N-terminus of VP5 alter its interaction with the scaffold proteins of herpes simplex virus type 1. AB - During the assembly process of herpes simplex virus type 1 capsids, there is an essential interaction between the C-terminal tail of the scaffold proteins (22a and 21) and the major capsid protein (VP5). Recent studies of spontaneous revertant viruses that overcome a blocked maturation cleavage site of the scaffold proteins have shown that the N-terminus of VP5 is important for this interaction. One of the revertant viruses, PR7, encodes a second-site mutation at residue 69 of VP5 which unlike wild-type VP5 fails to interact with 22a and thus gives white colonies in the yeast two-hybrid assay. In the present study a small DNA fragment, encoding residues 1 to 85 of wild-type and PR7 VP5, was mutagenized using error-prone PCR. Mutagenized DNA was used in the yeast two-hybrid assay to identify mutations in wild-type VP5 that resulted in loss of 22a binding (white colonies), or in PR7 VP5 that resulted in a gain of function (blue colonies). For the loss of function experiments, using KOS VP5, a row of eight thymidine nucleotides (codons 37-40) resulted in many frameshift mutations, which led us to terminate the study without reaching a statistically significant result. For the PR7 experiment, 30 clones were identified that had single amino acid substitutions, and these mutations were localized to amino acids 27-45 and 63-84 of VP5. The most frequent mutation was a reversion back to wild-type. The next most frequent were E28K and N63S, and these gave the highest beta-galactosidase enzyme activities (indicative of PR7VP5-22a interaction), 30 and 20% of wild type, respectively. When E28K and N63S were transferred into the wild-type VP5 background, that is, in the absence of the PR7 mutation, they gave rise to different phenotypes. The E28K mutation lost its ability to interact with the scaffold proteins as judged by this assay. Therefore, it may be acting as a compensatory mutation whose phenotype is only expressed in the presence of the original PR7 mutation. However, the N63S mutation in the wild-type VP5 background increased the interaction, as judged by the beta-galactosidase activity, by a factor of 9 relative to when the PR7 mutation was present. Even more surprising, in the absence of the PR7 mutation the enzyme activity was still greater, by a factor of 2, than that observed for wild-type VP5. This study provides further evidence that the N-terminus of VP5 is in intimate association with the C terminus of the scaffold proteins. PMID- 11384230 TI - Simple selection criteria for drug-like chemical matter. AB - A simple pharmacophore point filter has been developed that discriminates between drug-like and nondrug-like chemical matter. It is based on the observation that nondrugs are often underfunctionalized. Therefore, a minimum count of well defined pharmacophore points is required to pass the filter. The application of the filter results in 66-69% of subsets of the MDDR database to be classified as drug-like. Furthermore, 61-68% of subsets of the CMC database are classified as drug-like. In contrast, only 36% of the ACD are found to be drug-like. While these results are not quite as good as those obtained with recently described neural net approaches, the method used here has clear advantages. In contrast to a neural net approach and also in contrast to decision tree methods described recently, the pharmacophore filter has been developed by using "chemical wisdom" that is unbiased from fitting the structural content of specific drug databases to prediction models. Similar to decision tree methods, the pharmacophore point filter provides a detailed structural reason for the classification of each molecule as drug or nondrug. The pharmacophore point filter results are compared to neural net filter results. A statistically significant overlap between compounds recognized as drug-like validates both approaches. The pharmacophore point filter complements neural net approaches as well as property profiling approaches used as drug-likeness filters in compound library analysis and design. PMID- 11384231 TI - 2-(2-Oxo-1,4-dihydro-2H-quinazolin-3-yl)- and 2-(2,2-dioxo-1,4-dihydro-2H 2lambda6-benzo[1,2,6]thiadiazin-3-yl)-N-hydroxy-acetamides as potent and selective peptide deformylase inhibitors. AB - Potent, selective, and structurally new inhibitors of the Fe(II) enzyme Escherichia coli peptide deformylase (PDF) were obtained by rational optimization of the weakly binding screening hit (5-chloro-2-oxo-1,4-dihydro-2H-quinazolin-3 yl)-acetic acid hydrazide (1). Three-dimensional structural information, gathered from Ni-PDF complexed with 1, suggested the preparation of two series of related hydroxamic acid analogues, 2-(2-oxo-1,4-dihydro-2H-quinazolin-3-yl)-N-hydroxy acetamides (A) and 2-(2,2-dioxo-1,4-dihydro-2H-2lambda(6)-benzo[1,2,6]thiadiazin 3-yl)-N-hydroxy-acetamides (B), among which potent PDF inhibitors (37, 42, and 48) were identified. Moreover, two selected compounds, one from each series, 36 and 41, showed good selectivity for PDF over several endoproteases including matrix metalloproteases. However, these compounds showed only weak antibacterial activity. PMID- 11384232 TI - Identification of a putative binding site for [2',5'-bis-O-(tert butyldimethylsilyl)-beta-D-ribofuranosyl]-3'-spiro-5''-(4''-amino-1'',2'' oxathiole-2'',2''-dioxide)thymine (TSAO) derivatives at the p51-p66 interface of HIV-1 reverse transcriptase. AB - A binding site for TSAO-m(3)T at the interface between the p66 and p51 subunits of HIV-1 reverse transcriptase (RT) and distinct from that of "classical" HIV-1 non-nucleoside inhibitors is proposed. The feasibility of the binding mode was assessed by carrying out nanosecond molecular dynamics simulations for the complexes of TSAO-m(3)T with reduced models of both the wild-type enzyme and a more sensitive R172A mutant. The molecular model is in agreement with a previous proposal, with known structure-activity and mutagenesis data for this unique class of inhibitors, and also with recent biochemical evidence indicating that TSAO analogues can affect enzyme dimerization. The relative importance of residues involved in dimer formation and TSAO-RT complex stabilization was assessed by a combination of surface area accessibility, molecular mechanics, and continuum electrostatics calculations. A structure-based modification introduced into the lead compound yielded a new derivative with improved antiviral activity. PMID- 11384233 TI - 2-Amino-6-arylsulfonylbenzonitriles as non-nucleoside reverse transcriptase inhibitors of HIV-1. AB - A series of 2-amino-5-arylthiobenzonitriles (1) was found to be active against HIV-1. Structural modifications led to the sulfoxides (2) and sulfones (3). The sulfoxides generally showed antiviral activity against HIV-1 similar to that of 1. The sulfones, however, were the most potent series of analogues, a number having activity against HIV-1 in the nanomolar range. Structural-activity relationship (SAR) studies suggested that a meta substituent, particularly a meta methyl substituent, invariably increased antiviral activities. However, optimal antiviral activities were manifested by compounds where both meta groups in the arylsulfonyl moiety were substituted and one of the substituents was a methyl group. Such a disubstitution led to compounds 3v, 3w, 3x, and 3y having IC50 values against HIV-1 in the low nanomolar range. When gauged for their broad spectrum antiviral activity against key non-nucleoside reverse transcriptase inhibitor (NNRTI) related mutants, all the di-meta-substituted sulfones 3u-z and the 2-naphthyl analogue 3ee generally showed single-digit nanomolar activity against the V106A and P236L strains and submicromolar to low nanomolar activity against strains E138K, V108I, and Y188C. However, they showed a lack of activity against the K103N and Y181C mutant viruses. The elucidation of the X-ray crystal structure of the complex of 3v (739W94) in HIV-1 reverse transcriptase showed an overlap in the binding domain when compared with the complex of nevirapine in HIV 1 reverse transcriptase. The X-ray structure allowed for the rationalization of SAR data and potencies of the compounds against the mutants. PMID- 11384234 TI - 3D-QSAR model of flavonoids binding at benzodiazepine site in GABAA receptors. AB - With flavone as a structural template, three-dimensional quantitative structure activity relationship (3D-QSAR) studies and ab initio calculations were performed on a series of flavonoids. A reasonable pharmacophore model was built through CoMFA, CoMSIA, and HQSAR analyses and electrostatic potential calculations. A plausible binding mode for flavonoids with GABA(A) receptors was rationalized. On the basis of the commonly recognized binding site, the specific S1 and S2 subsites relating to substituent positions were proposed. The different binding affinities could be explained according to the frontier orbitals and electrostatic potential (ESP) maps. The ESP could be used as a novel starting point for designing more selective BZ-binding-site ligands. PMID- 11384235 TI - An optimized protein kinase C activating diacylglycerol combining high binding affinity (Ki) with reduced lipophilicity (log P). AB - A small, focused combinatorial library encompassing all possible permutations of acyl branched alkyl chains-small and large, saturated and unsaturated-was generated from the active diacylglycerol enantiomer (S-DAG) to help identify the analogue with the highest binding affinity (lowest Ki) for protein kinase C (PK C) combined with the minimum lipophilicity (log P). The selected ligand (3B) activated PK-C more effectively than sn-1,2-dioctanoylglycerol (diC8) despite being 1.4 log units more hydrophilic. Compound 3B indeed represents the most potent, hydrophilic DAG ligand to date. With the help of a green fluorescent protein (GFP)-tagged PK-Calpha, 3B was able to translocate the full length protein to the membrane with an optimal dose of 100 microM in CHO-K1 cells, while diC8 failed to achieve translocation even at doses 3-fold higher. Molecular modeling of 3B into an empty C1b domain of PK-Cdelta clearly showed the existence of a preferred binding orientation. In addition, molecular dynamic simulations suggest that binding discrimination could result from a favorable van der Waals (VDW) interaction between the large, branched sn-1 acyl group of 3B and the aromatic rings of Trp252 (PK-Cdelta) or Tyr252 (PK-Calpha). The DAG analogue of 3B in which the acyl groups are reversed (2C) showed a decrease in binding affinity reflecting the capacity of PK-C to effectively discriminate between alternative orientations of the acyl chains. PMID- 11384236 TI - Radioiodinated styrylbenzenes and thioflavins as probes for amyloid aggregates. AB - We report for the first time that small molecule-based radiodiodinated ligands, showing selective binding to Abeta aggregates, cross the intact blood-brain barrier by simple diffusion. Four novel ligands showing preferential labeling of amyloid aggregates of Abeta(1-40) and Abeta(1-42) peptides, commonly associated with plaques in the brain of people with Alzheimer's disease (AD), were developed. Two 125I-labeled styrylbenzenes, (E,E)-1-iodo-2,5-bis(3 hydroxycarbonyl-4-hydroxy)styrylbenzene, 12 (ISB), and (E,E)-1-iodo-2,5-bis(3 hydroxycarbonyl-4-methoxy)styrylbenzene, 13 (IMSB), and two 125I-labeled thioflavins, 2-[4'-(dimethylamino)phenyl]-6-iodobenzothiazole, 18a (TZDM), and 2 [4'-(4''-methylpiperazin-1-yl)phenyl]-6-iodobenzothiazole, 18b (TZPI), were prepared at a high specific activity (2200 Ci/mmol). In vitro binding studies of these ligands showed excellent binding affinities with Kd values of 0.08, 0.13, 0.06, and 0.13 nM for aggregates of Abeta(1-40) and 0.15, 0.73, 0.14, and 0.15 nM for aggregates of Abeta(1-42), respectively. Interestingly, under a competitive binding assaying condition, different binding sites on Abeta(1-40) and Abeta(1 42) aggregates, which are mutually exclusive, were observed for styrylbenzenes and thioflavins. Autoradiography studies of postmortem brain sections of a patient with Down's syndrome known to contain primarily Abeta(1-42) aggregates in the brain showed that both [(125)I]18a and [125I]18b labeled these brain sections, but [125I]13, selective for Abeta(1-40) aggregates, exhibited very low labeling of the comparable brain section. Biodistribution studies in normal mice after an iv injection showed that [125I]18a and [(125)I]18b exhibited excellent brain uptake and retention, the levels of which were much higher than those of [125I]12 and [125I]13. These findings strongly suggest that the new radioiodinated ligands, [125I]12 (ISB), [125I]13 (IMSB), [125I]18a (TZDM), and [125I]18b (TZPI), may be useful as biomarkers for studying Abeta(1-40) as well as Abeta(1-42) aggregates of amyloidogenesis in AD patients. PMID- 11384237 TI - Soluble 2-substituted aminopyrido[2,3-d]pyrimidin-7-yl ureas. Structure-activity relationships against selected tyrosine kinases and exploration of in vitro and in vivo anticancer activity. AB - In continuing our search for medicinal agents to treat proliferative diseases, we have discovered 2-substituted aminopyrido[2,3-d]pyrimidin-7-yl ureas as a novel class of soluble, potent, broadly active tyrosine kinase (TK) inhibitors. An efficient route was developed that enabled the synthesis of a wide variety of analogues with substitution on several positions of the template. From the lead structure 1, several series of analogues were made that examined the C-6 aryl substituent, a variety of water solublizing substitutents at the C-2 position, and urea or other acyl functionality at the N-7 position. Compounds of this series were competitive with ATP and displayed submicromolar to low nanomolar potency against a panel of TKs, including receptor (platelet-derived growth factor, PDGFr; fibroblast growth factor, FGFr;) and nonreceptor (c-Src) classes. Several of the most potent compounds displayed submicromolar inhibition of PDGF mediated receptor autophosphorylation in rat aortic vascular smooth muscle cells and low micromolar inhibition of cellular growth in five human tumor cell lines. One of the more thoroughly evaluated members, 32, with IC50 values of 0.21 microM (PDGFr), 0.049 microM (bFGFr), and 0.018 microM (c-Src), was evaluated in in vivo studies against a panel of five human tumor xenografts, with known and/or inferred dependence on the EGFr, PDGFr, and c-Src TKs. Compound 32 produced a tumor growth delay of 14 days against the Colo-205 colon xenograft model. PMID- 11384238 TI - Experimental and computational screening models for the prediction of intestinal drug absorption. AB - The aim of this study was to devise experimental protocols and computational models for the prediction of intestinal drug permeability. Both the required experimental and computational effort and the accuracy and quality of the resulting predictions were considered. In vitro intestinal Caco-2 cell monolayer permeabilities were determined both in a highly accurate experimental setting (Pc) and in a faster, but less accurate, mode (Papp). Computational models were built using four different principles for generation of molecular descriptors (atom counts, molecular mechanics calculations, fragmental, and quantum mechanics approaches) and were evaluated for their ability to predict intestinal membrane permeability. A theoretical deconvolution of the polar molecular surface area (PSA) was also performed to facilitate the interpretation of this composite descriptor and allow the calculation of PSA in a simplified and fast mode. The results indicate that it is possible to predict intestinal drug permeability from rather simple models with little or no loss of accuracy. A new, fast computational model, based on partitioned molecular surface areas, that predicts intestinal drug permeability with an accuracy comparable to that of time consuming quantum mechanics calculations is presented. PMID- 11384239 TI - Solid-phase synthesis of a nonpeptide RGD mimetic library: new selective alphavbeta3 integrin antagonists. AB - The solid-phase synthesis of a low molecular weight RGD mimetic library is described. Activities of the compounds in inhibiting the interaction of ligands, vitronectin and fibrinogen, with isolated immobilized integrins alphavbeta3 and alphaIIbbeta3 were determined in a screening assay. Highly active and selective nonpeptide alphavbeta3 integrin antagonists with regard to orally bioavailability were developed, based on the aza-glycine containing lead compound 1. An important variation is the substitution of the aspartic amide of 1 by an aromatic residue. Furthermore, different guanidine mimetics have been incorporated to improve the pharmacokinetic profile. Exchange of the beta-amino acid NH by a methylene moiety in one set of RGD mimetics leads to the azacarba analogue compounds representing a novel peptidomimetic approach, which should increase the metabolic stability. PMID- 11384240 TI - Structure-activity relationships of 1,4-dihydro-(1H,4H)-quinoxaline-2,3-diones as N-methyl-D-aspartate (glycine site) receptor antagonists. 1. Heterocyclic substituted 5-alkyl derivatives. AB - A series of 6,7-dichloro-1,4-dihydro-(1H, 4H)-quinoxaline-2,3-diones (1-17) were prepared in which the 5-position substituent was a heterocyclylmethyl or 1 (heterocyclyl)-1-propyl group. Structure-activity relationships were evaluated where binding affinity for the glycine site of the N-methyl-D-aspartate (NMDA) receptor was measured using the specific radioligand [3H]-L-689,560, and functional antagonism was demonstrated by inhibition of NMDA-induced depolarizations of rat cortical wedges. The ability to prevent NMDA-induced hyperlocomotion in mice in vivo was measured for selected compounds. Binding affinity increased significantly if the heterocyclic group, e.g. 1,2,3-triazol-1 yl could participate in accepting a hydrogen bond from the receptor. It was difficult to obtain compounds with adequate aqueous solubility and strategies to improve it were investigated. The most potent compound in this series, 6,7 dichloro-5-[1-(1,2,4-triazol-4-yl)propyl]-1,4-dihydro-(1H, 4H)-quinoxaline-2,3 dione (17) (binding IC50 = 2.6 nM; cortical wedge EC50 = 90 nM), inhibited NMDA induced hyperlocomotion in mice (6/9 protected at 20 mg/kg iv). Pharmacokinetic parameters, including extent of brain penetration, for 11 and 17 are reported. PMID- 11384241 TI - Investigations of new lead structures for the design of selective estrogen receptor modulators. AB - Heterocyclic derivatives of (R,S)/(S,R)-1-(2-chloro-4-hydroxyphenyl)-2-(2,6 dichloro-4-hydroxyphenyl)ethylenediamine (L1) were synthesized and tested for estrogen receptor binding. The selection of the heterocycles was based on theoretical consideration. (2R,3S)/(2S,3R)-2-(2-Chloro-4-hydroxyphenyl)-3-(2,6 dichloro-4-hydroxyphenyl)piperazine 2, (4R,5S)/(4S,5R)-4-(2-chloro-4 hydroxyphenyl)-5-(2,6-dichloro-4-hydroxyphenyl)-2-imidazoline 3, and 4-(2-chloro 4-hydroxyphenyl)-5-(2,6-dichloro-4-hydroxyphenyl)imidazole 4 possess a spatial structure with neighboring aromatic rings as is realized in hormonally active [1,2-diphenylethylenediamine]platinum(II) complexes. The 1,2-diphenylethane pharmacophor, however, cannot adapt an antiperiplanar conformation to interact with the estrogen receptor (ER) comparable to synthetic (e.g., diethylstilbestrol (DES)) or steroidal (e.g., estradiol (E2)) estrogens. Due to the different spatial structures, the heterocycles cause only a marginal displacement of E2 from its binding site (relative binding affinity (RBA) < 0.1%). Nevertheless, unequivocally ER mediated gene activation was verified on the MCF-7-2a cell line. Imidazoline 3 as the most active compound reached the maximum effect of E2 (100% activation) in a concentration of 5 x 10(-7) M, while piperazine 2 and imidazole 4 activate luciferase expression only in a small but significant amount of 20% and 27%, respectively. We therefore assigned these heterocyclic compounds to a second class of hormones (type-II-estrogens), which are attached at the ER at different amino acids than DES or E2 (type-I-estrogens). PMID- 11384242 TI - Structure-activity studies for a novel series of bicyclic substituted hexahydrobenz[e]isoindole alpha1A adrenoceptor antagonists as potential agents for the symptomatic treatment of benign prostatic hyperplasia. AB - In search of a uroselective alpha1A subtype selective antagonist, a novel series of 6-OMe hexahydrobenz[e]isoindoles attached to a bicyclic heterocyclic moiety via a two-carbon linker was synthesized. It was found that in contrast to the previously described series of tricyclic heterocycles,(1) this bicyclic series has very specific requirements for the heterocyclic attachments. The most important structural features contributing to the alpha1A/alpha1B selectivity of these compounds were identified. In vitro functional assays for the alpha1 adrenoceptor subtypes were used to further characterize the most selective compounds, and in vivo models of vascular vs prostatic tone were used to assess uroselectivity. Compound 48 showed the highest degree of selectivity in the radioligand binding assays (56-fold), in the in vitro functional tests (80-fold), and for in vivo prostate selectivity (960-fold). PMID- 11384243 TI - Aromatic quinolinecarboxamides as selective, orally active antibody production inhibitors for prevention of acute xenograft rejection. AB - The prevention of xenograft rejection is substantially dependent on inhibiting antibodies (Ab) produced by B-cells independently of T-cell signals (TI-1). Due to their ubiquitous biochemical mechanisms of action, the immunosuppressants currently employed not only fail to discriminate between B- and T-cells but also have a narrow therapeutic window and, thus, their prolonged use in complex immunosuppressive regimens is problematic. By capitalizing on the target enzyme bound (DHODH) structure 1b of one of these compounds, leflunomide, and modulating part of its multiple mechanisms of action to gain selectivity, the quinoline-8 carboxamide 3 was designed as a potentially weak enzyme inhibitor but effective immunosuppressant. Compound 3 fulfilled the mechanistic criteria set and had 10 fold B-cell over T-cell selectivity. Its pyridyl analogue 4 was found to be a highly potent and selective B-cell immunosuppressant with a 75-fold selectivity for B- over T-cells (as judged by the MLR data) and no general cytotoxicity at concentrations up to 160-fold higher than those required to inhibit B-cells. In the mouse, 4 effectively blocked TI-1 Ab production and suppressed Ab-mediated xenograft rejection in a xenotransplantation model under a once-daily dosing regimen, with efficacy down to 0.3 mg/kg/day po. These are the first data demonstrating the feasibility of the development of drugs specific for impeding Ab production. PMID- 11384244 TI - Synthesis, antifolate, and antitumor activities of classical and nonclassical 2 amino-4-oxo-5-substituted-pyrrolo[2,3-d]pyrimidines. AB - Classical and nonclassical isosteric C8-N9 bridged analogues of the multitargeted antifolate LY231514 were synthesized as inhibitors of thymidylate synthase (TS), dihydrofolate reductase (DHFR), and as antitumor and antiopportunistic infection agents. The syntheses of the analogues were accomplished by reductive amination of the appropriate anilines with 2-amino-4-oxo-5-cyanopyrrolo[2,3-d]pyrimidine (28) followed by saponification of the ethyl esters, for the classical analogue 6. The N9-methyl analogues were obtained from the N9-H precursors by reductive methylation. In general, the nonclassical compounds 7-17 were similar in potency to TMP against Toxoplasma gondii DHFR, with selectivity ratios greater than 38 and 21 for 11 and 16, respectively. These compounds were poor inhibitors of Pneumocystis carinii DHFR and rat liver DHFR. The nonclassical analogues were also inactive against TS. The classical analogue 6 was a marginal inhibitor of isolated human TS (IC50 = 46 microM) and of human DHFR (IC50 = 10 microM), however, it was a potent inhibitor of the growth of two human head and neck squamous cell carcinoma cell lines and of CCRF-CEM human lymphoblastic leukemia cells in culture and was similar to LY231514 against ZR-75-1 human breast carcinoma cell line. Evaluation of 6 against MTX-resistant sublines indicated that DHFR is not the major target of 6. Metabolite protection studies of the growth inhibitory activity of 6 suggest that TS is a major target of this drug and that polyglutamyl forms of 6 may serve as the intracellular TS inhibitors. These studies also suggest that 6 has a site of action in addition to sites in the folate pathway. PMID- 11384245 TI - Synthesis and cytotoxic activity of 7-oxo-7H-dibenz[f,ij]isoquinoline and 7-oxo 7H-benzo[e]perimidine derivatives. AB - A series of 7-oxo-7H-dibenz[f,ij]isoquinoline and 7-oxo-7H-benzo[e]perimidines bearing cationic side chains were prepared from aminoanthraquinones. The perimidines were prepared from 1-aminoanthraquinone by initial condensation with urea or dimethylacetamide. A series of 2-, 4-, 8-, and 11-carboxy derivatives of the dibenzisoquinolines were prepared from aminoanthraquinonecarboxylic acids. The cationic derivatives were prepared from these via amide, amine, or methylene linkers to study the effects of side chain positioning on biological activity. Within the series of carboxamide-linked compounds, the order of increasing cytotoxicity was 8- < 4- < 2- < 11-. The 2- and 4-carboxamides showed substantial growth delays against in vivo subcutaneous colon 38 tumors in mice, but the 11 carboxamide had curative activity in this refractory model and is being investigated further. PMID- 11384246 TI - Potent and selective nonpeptide inhibitors of caspases 3 and 7. AB - 5-Dialkylaminosulfonylisatins have been identified as potent, nonpeptide inhibitors of caspases 3 and 7. The most active compound within this series (34) inhibited caspases 3 and 7 in the 2-6 nM range and exhibited approximately 1000 fold selectivity for caspases 3 and 7 versus a panel of five other caspases (1, 2, 4, 6, and 8) and was at least 20-fold more selective versus caspase 9. Sequence alignments of the active site residues of the caspases strongly suggest that the basis of this selectivity is due to binding in the S2 subsite comprised of residues Tyr204, Trp206, and Phe256 which are unique to caspases 3 and 7. These compounds inhibit apoptosis in three cell-based models: human Jurkat T cells, human chondrocytes, and mouse bone marrow neutrophils. PMID- 11384247 TI - Metabolism of sirolimus and its derivative everolimus by cytochrome P450 3A4: insights from docking, molecular dynamics, and quantum chemical calculations. AB - A combination of quantum chemical calculations and molecular simulations (DOCKing and molecular dynamics) is used to investigate the metabolism of sirolimus (rapamycin) and its derivative everolimus (SDZ-RAD) by cytochrome P450 3A4. Both molecules are drugs with high immunosuppressive activity. Our calculations yield qualitative predictions of the regiospecificities of the hydroxylations and O dealkylations occurring in these two substrates which are in good agreement with recent experimental results. An analysis of the modeled enzyme-substrate interactions allows us to rationalize the reduced metabolic activity of the larger substrate everolimus compared to sirolimus. Moreover, our simulations suggest that hydrogen donor functionalities close to the metabolic site are important for anchoring the substrate at the active center of the enzyme. In particular, we predict that replacing one hydroxyl group by a fluorine atom should considerably suppress the major metabolic reaction in sirolimus, 39-O demethylation. PMID- 11384248 TI - Preparation and properties of S-nitroso-L-cysteine ethyl ester, an intracellular nitrosating agent. AB - In this report, a protocol for the preparation of the hydrochloride of S-nitroso L-cysteine ethyl ester (SNCEE.HCl; 2) is presented. The synthesis of 2 has been targeted because S-nitroso-L-cysteine (SNC; 2b), which is extensively used for trans-S-nitrosation of thiol-containing proteins, has a limited ability of crossing cellular membranes. The nitrosothiol 2 was prepared via direct S nitrosation of the hydrochloride of L-cysteine ethyl ester (CEE.HCl; 1a) with ethyl nitrite. 2 is relatively stable in crystal form and when neutralized to SNCEE (2a) in aqueous solutions treated with chelators of metal ions. Traces of metal ions, however, triggered the decomposition of 2a to nitric oxide and a S centered radical, which were detected by ESR spectrometry. In contrast to 2b, 2a is a lipophilic compound that was taken up by human neutrophils. The latter process was paralleled by inhibition of the NADPH oxidase-dependent generation of superoxide anion radicals, presumably via reaction(s) of intracellular trans-S nitrosation. Intracellular accumulation of S-nitrosothiols was observed with 2a but not with 2b. It is expected that the use of 2a will be advantageous when intracellular reactions of trans-S-nitrosation are to be studied. PMID- 11384250 TI - Simultaneous minimum-uncertainty measurement of discrete-valued complementary observables. AB - We have made the first experimental demonstration of the simultaneous minimum uncertainty product between two complementary observables for a two-state system (a qubit). A partially entangled two-photon state, where each of the photons carries (partial) information of the initial state, was used to perform such a measurement. PMID- 11384251 TI - Hidden-variable theorems for real experiments. AB - It has recently been questioned whether the Kochen-Specker theorem is relevant to real experiments, which by necessity only have finite precision. We give an affirmative answer to this question by showing how to derive hidden-variable theorems that apply to real experiments, so that noncontextual hidden variables can indeed be experimentally disproved. The essential point is that for the derivation of hidden-variable theorems one does not have to know which observables are really measured by the apparatus. Predictions can be derived for observables that are defined in an entirely operational way. PMID- 11384252 TI - Entanglement and extreme spin squeezing. AB - For any mean value of a Cartesian component of a spin vector we identify the smallest possible uncertainty in any of the orthogonal components. The corresponding states are optimal for spectroscopy and atomic clocks. We show that the results for different spin J can be used to identify entanglement and to quantify the depth of entanglement in systems with many particles. With the procedure developed in this Letter, collective spin measurements on an ensemble of particles can be used as an experimental proof of multiparticle entanglement. PMID- 11384253 TI - Experimental demonstration of four-photon entanglement and high-fidelity teleportation. AB - We experimentally demonstrate observation of highly pure four-photon GHZ entanglement produced by parametric down-conversion and a projective measurement. At the same time this also demonstrates teleportation of entanglement with very high purity. Not only does the achieved high visibility enable various novel tests of quantum nonlocality, it also opens the possibility to experimentally investigate various quantum computation and communication schemes with linear optics. Our technique can, in principle, be used to produce entanglement of arbitrarily high order or, equivalently, teleportation and entanglement swapping over multiple stages. PMID- 11384254 TI - Quantum dynamics of strongly interacting boson systems: atomic beam splitters and coupled Bose-Einstein condensates. AB - An effective boson Hamiltonian applicable to atomic beam splitters, coupled Bose Einstein condensates, and optical lattices can be made exactly solvable by including all n-body interactions. The model can include an arbitrary number of boson components. In the strong interaction limit the model becomes a quantum phase model, which also describes a tight-binding lattice particle. Through exact results for dynamic correlation functions, it is shown how the previous weak interaction dynamics of these systems are extended to strong interactions, now becoming relevant in the experiments. The effect of the number of boson components is also analyzed. PMID- 11384255 TI - Stationary states of a rotating Bose-Einstein condensate: routes to vortex nucleation. AB - Using a focused laser beam we stir a 87Rb Bose-Einstein condensate confined in a magnetic trap. We observe that the steady states of the condensate correspond to an elliptic cloud, stationary in the rotating frame. These steady states depend nonlinearly on the stirring parameters (amplitude and frequency), and various solutions can be reached experimentally depending on the path followed in this parameter space. These states can be dynamically unstable and we observe that such instabilities lead to vortex nucleation in the condensate. PMID- 11384256 TI - Superfluid and dissipative dynamics of a Bose-Einstein condensate in a periodic optical potential. AB - We create Bose-Einstein condensates of 87Rb in a static magnetic trap with a superimposed blue-detuned 1D optical lattice. By displacing the magnetic trap center we are able to control the condensate evolution. We observe a change in the frequency of the center-of-mass oscillation in the harmonic trapping potential, in analogy with an increase in effective mass. For fluid velocities greater than a local speed of sound, we observe the onset of dissipative processes up to full removal of the superfluid component. A parallel simulation study visualizes the dynamics of the Bose-Einstein condensate and accounts for the main features of the observed behavior. PMID- 11384257 TI - BPS states in M theory and twistorial constituents. AB - We provide a complete algebraic description of Bogomol'nyi-Prasad-Sommerfield (BPS) states in M theory in terms of primary constituents that we call BPS preons. We argue that any BPS state preserving k of the 32 supersymmetries is a composite of (32-k) BPS preons. In particular, the BPS states corresponding to the basic M2 and M5 branes are composed of 16 BPS preons. By extending the M algebra to a generalized D = 11 conformal superalgebra osp(1/64) we relate the BPS preons with its fundamental representation, the D = 11 supertwistors. PMID- 11384258 TI - Numerical analysis of the double scaling limit in the string type IIB matrix model. AB - The bosonic IIB matrix model is studied using a numerical method. This model contains the bosonic part of the IIB matrix model conjectured to be a nonperturbative definition of the type IIB superstring theory. The large N scaling behavior of the model is shown performing a Monte Carlo simulation. The expectation value of the Wilson loop operator is measured and the string tension is estimated. The numerical results show the prescription of the double scaling limit. PMID- 11384259 TI - Metastable ringlike semitopological solitons. AB - We show the existence of new stable ringlike localized scalar field configurations whose stability is due to a combination of topological and nontopological charges. In that sense these defects may be called semitopological. These rings are Noether charged and also carry Noether current (they are superconducting). They are local minima of the energy in scalar field theories with an unbroken U(1) global symmetry. We obtain numerical solutions of the field configuration corresponding to large rings and derive virial theorems demonstrating their stability. We also derive the minimum energy field configurations in 3D and simulate the evolution of a finite size Q ring on a three dimensional lattice. PMID- 11384260 TI - Do electroweak precision data and Higgs-mass constraints rule out a scalar bottom quark with mass of order 5 GeV? AB - We study the implications of a scalar bottom quark, with a mass of O (5 GeV), within the minimal supersymmetric standard model. Light sbottoms may naturally appear for large tan(beta) and, depending on the decay modes, may have escaped experimental detection. We show that a light sbottom cannot be ruled out by electroweak precision data and the bound on the lightest CP-even Higgs-boson mass. We infer that a light b scenario requires a relatively light scalar top quark whose mass is typically about the top-quark mass. In this scenario the lightest Higgs boson decays predominantly into b pairs and obeys the mass bound m(h) less, similar 123 GeV. PMID- 11384265 TI - Rotations in isospace: a doorway to the understanding of neutron-proton superfluidity in N = Z nuclei. AB - The T = 2 excitations in even-even N = Z nuclei are calculated within the isospin cranked mean-field approach. The response of pairing correlations to rotation in isospace is investigated. Whereas the isovector pairing rather modestly modifies the single-particle moment of inertia in isospace, the isoscalar pairing strongly reduces its value. This reduction of the isomoments of inertia with respect to its rigid body value is a strong indicator of collective isoscalar pairing correlations. These results are further generalized yielding beautiful analogies between the role of isovector pairing for the case of spatial rotations and the role of isoscalar pairing for the case of isorotations. PMID- 11384264 TI - Energy loss of fast quarks in nuclei. AB - We report an analysis of the nuclear dependence of the yield of Drell-Yan dimuons from the 800 GeV/c proton bombardment of 2H, C, Ca, Fe, and W targets. Employing a new formulation of the Drell-Yan process in the rest frame of the nucleus, this analysis examines the effect of initial-state energy loss and shadowing on the nuclear-dependence ratios versus the incident proton's momentum fraction and dimuon effective mass. The resulting energy loss per unit path length is -dE/dz = 2.32+/-0.52+/-0.5 GeV/fm. This is the first observation of a nonzero energy loss of partons traveling in a nuclear environment. PMID- 11384266 TI - Nuclear fragmentation: sampling the instabilities of binary systems. AB - We derive stability conditions of asymmetric nuclear matter (ANM) and discuss the relation to mechanical and chemical instabilities of general two-component systems. We show that the chemical instability may appear as an instability of the system against isoscalarlike rather than isovectorlike fluctuations if the interaction between the two constituent species has an attractive character as in the case of ANM. This leads to a new kind of liquid-gas phase transition, of interest for fragmentation experiments with radioactive beams. PMID- 11384261 TI - Study of tau decays to six pions and a neutrino. AB - The tau decays to six-pion final states have been studied with the CLEO detector at the Cornell Electron Storage Ring. The measured branching fractions are B(tau( )-->2pi(-)pi(+)3pi(0)nu(tau)) = (2.2+/-0.3+/-0.4)x10(-4) and B(tau(-)-->3pi( )2pi(+)pi(0)nu(tau)) = (1.7+/-0.2+/-0.2)x10(-4). A search for substructure in these decays shows that they are saturated by intermediate states with eta or omega mesons. We present the first observation of the decay tau(-)-->2pi( )pi(+)omega(nu)tau and the branching fraction is measured to be (1.2+/-0.2+/ 0.1)x10(-4). The measured branching fractions are in good agreement with the isospin expectations but somewhat below the conserved-vector-current predictions. PMID- 11384267 TI - Alternative to the hyperfine-change-collision interpretation for the behavior of magneto-optical-trap losses at low light intensity. AB - Usually, the large trap loss rates observed in MOTs at the low light intensity regime have been associated with hyperfine change collisions (HCC). We propose an alternative mechanism to explain the sudden raise up of trap loss rates at low intensity without relying on HCC. Using the Gallagher-Pritchard model [Phys. Rev. Lett. 63, 957 (1989)] together with an intensity dependent escape velocity, we were able to reproduce qualitatively well some existing experimental results, including recent observations by Nesnidal et al. [Phys. Rev. A 62, 030701(R) (2000)]. This result reopens the discussion in order to better understand the physical mechanisms and their actual contribution to the trap losses. PMID- 11384268 TI - Photoelectron imaging on time-dependent molecular alignment created by a femtosecond laser pulse. AB - Rotational wave packet revivals on an excited electronic state have been measured by femtosecond time-resolved photoelectron imaging for the first time. The first full revival at 82 ps of S1 (n,pi*) pyrazine was clearly observed in the time dependencies of the photoelectron intensity and the photoelectron angular distribution (PAD). The PAD, measured for laser aligned pyrazine, clearly reflects the different characters of pi* and 3s molecular orbitals. PMID- 11384269 TI - Anionic photofragmentation of CO: a selective probe of core-level resonances. AB - Anion-yield spectroscopy using x rays is shown to be a selective probe of molecular core-level processes, providing unique experimental verification of shape resonances. For CO, partial anion and cation yields are presented for photon energies near the C K edge. The O- yield exhibits features above threshold related only to doubly excited states, in contrast to cation yields which also exhibit pronounced structure due to the well-known sigma* shape resonance. Because the shape resonance is completely suppressed for O-, anion spectroscopy thus constitutes a highly selective probe, yielding information unobtainable with absorption or electron spectroscopy. PMID- 11384270 TI - Quantum-mechanical analysis of a longitudinal Stern-Gerlach effect. AB - We present the results of a rigorous quantum-mechanical calculation of the propagation of electrons through an inhomogeneous magnetic field with axial symmetry. A complete spin polarization of the beam is demonstrated assuming that a Landau eigenstate can be inserted into the field. This is in contrast with the semiclassical situation, where the spin splitting is blurred. PMID- 11384271 TI - Polarization instabilities in a two-photon laser. AB - We describe the operating characteristics of a new type of quantum oscillator that is based on a two-photon stimulated emission process. This two-photon laser consists of spin-polarized and laser-driven 39K atoms placed in a high-finesse transverse-mode-degenerate optical resonator and produces a beam with a power of approximately 0.2 microW at a wavelength of 770 nm. We observe complex dynamical instabilities of the state of polarization of the two-photon laser, which are made possible by the atomic Zeeman degeneracy. We conjecture that the laser could emit polarization-entangled twin beams if this degeneracy is lifted. PMID- 11384263 TI - Observation of new states decaying into Lambda+(c)pi(-)pi(+). AB - Using 13.7 fb(-1) of data recorded by the CLEO detector at the Cornell Electron Storage Ring, we investigate the spectrum of charmed baryons which decay into Lambda+(c)pi(-)pi(+) and are more massive than the Lambda+(c)(2625) baryon. We find evidence for two new states: one is broad and has an invariant mass roughly 480 MeV above that of the Lambda+(c) baryon; the other is narrow with an invariant mass of 596+/-1+/-2 MeV above the Lambda+(c) mass. PMID- 11384272 TI - Entangled-state lithography: tailoring any pattern with a single state. AB - We demonstrate a systematic approach to Heisenberg-limited lithographic image formation using four-mode reciprocal binomial states. By controlling the exposure pattern with a simple bank of birefringent plates, any pixel pattern on a (N+1) x (N+1) grid, occupying a square with the side half a wavelength long, can be generated from a 2N-photon state. PMID- 11384273 TI - Phase transition in a radiation-matter interaction with recoil and collisions. AB - The standard model introduced to describe the collective atomic recoil of an ensemble of atoms interacting with a strong electromagnetic field has been here extended by the inclusion of collisions with a buffer gas. As a result, we find that in the thermodynamic limit the coherent emission of radiation exhibits a continuous phase transition upon increasing the pump intensity. The output laser field is strictly larger than 0 only above a critical value. We find that the transition is not associated with the onset of spatial ordering but rather with the onset of a synchronization between the polarization phase and spatial position. A coherence parameter is introduced to characterize the phase transition. PMID- 11384274 TI - Photon statistics of random lasers with resonant feedback. AB - We have measured the photon statistics of random lasers with resonant feedback. With an increase of the pump intensity, the photon number distribution in a single mode changes continuously from Bose-Einstein distribution at the threshold to Poisson distribution well above the threshold. The second-order correlation coefficient drops gradually from 2 to 1. By comparing the photon statistics of a random laser with resonant feedback and that of a random laser with nonresonant feedback, we illustrate very different lasing mechanisms for the two types of random lasers. PMID- 11384275 TI - Measurement of modulational instability gain of second-order nonlinear optical eigenmodes in a one-dimensional system. AB - We have investigated the amplification of a spatially periodic perturbation applied to a wide fundamental beam launched near phase matching for second harmonic generation in a lithium niobate film waveguide. We measured the gain coefficient for the modulational instability of quadratic eigenmodes as a function of periodicity, intensity, and wave-vector mismatch. Excellent agreement with theory was obtained. PMID- 11384276 TI - Microscopic selection of fluid fingering patterns. AB - We study the issue of the selection of viscous fingering patterns in the limit of small surface tension. Through detailed simulations of anisotropic fingering, we demonstrate conclusively that no selection independent of the small-scale cutoff (macroscopic selection) occurs in this system. Rather, the small-scale cutoff completely controls the pattern, even on short time scales, in accordance with the theory of microscopic solvability. We demonstrate that ordered patterns are dynamically selected only for not too small surface tensions. For extremely small surface tensions, the system exhibits chaotic behavior and no regular pattern is realized. PMID- 11384277 TI - Templating of thin films induced by dewetting on patterned surfaces. AB - The instability, dynamics, and morphological transitions of patterns in thin liquid films on chemically heterogeneous striped surfaces are investigated based on 3D nonlinear simulations. The film breakup is suppressed on some potentially destabilizing nonwettable sites when their spacing is below a characteristic length scale of the instability, lambda(h). The thin film pattern replicates the substrate surface energy pattern closely only when (i) the periodicity of substrate pattern lies between lambda(h) and 2lambda(h), and (ii) the stripe width is within a range bounded by a lower critical length, below which no heterogeneous rupture occurs, and an upper transition length above which complex morphological features unlike the substrate pattern are formed. PMID- 11384278 TI - Vertical pairing of identical particles suspended in the plasma sheath. AB - It is shown experimentally that vertical pairing of two identical microspheres suspended in the sheath of a radio-frequency (rf) discharge at low gas pressures (a few Pa) appears at a well-defined instability threshold of the rf power. The transition is reversible, but with significant hysteresis on the second stage. A simple model which uses measured microsphere resonance frequencies and takes into account, in addition to the Coulomb interaction between negatively charged microspheres, their interaction with positive-ion-wake charges, seems to explain the instability threshold quite well. PMID- 11384279 TI - Quiescent double barrier regime in the DIII-D tokamak. AB - A new sustained high-performance regime, combining discrete edge and core transport barriers, has been discovered in the DIII-D tokamak. Edge localized modes (ELMs) are replaced by a steady oscillation that increases edge particle transport, thereby allowing particle control with no ELM-induced pulsed divertor heat load. The core barrier resembles those usually seen with a low (L) mode edge, without the degradation often associated with ELMs. The barriers are separated by a narrow region of high transport associated with a zero crossing in the E x B shearing rate. PMID- 11384280 TI - Coexisting nematic and smectic-A phases in a twisted liquid-crystal cell. AB - Twisted homogeneously planar-aligned nematic liquid-crystal cells are cooled into the smectic- A phase. The expected defective structure does not form. Instead the cells still show good optical-guiding characteristics. Exploration of the cells using a half-leaky guided-mode arrangement reveals that the liquid-crystal phase separates into three or more regions. Adjacent to both the upper and lower boundaries is a region of highly twisted nematic liquid crystal. In the center of the cell is one or more homogeneous smectic- A regions with smectic layers normal to the cell surfaces, separated by twisted nematic. As the cell is cooled so the smectic- A regions grow in thickness with the nematic regions progressively thinning but with increased twist gradient. A theoretical model of these novel results is presented. PMID- 11384281 TI - Martensitic fcc-to-hcp transformation observed in xenon at high pressure. AB - Angle-resolved x-ray diffraction patterns of Xe to 127 GPa indicate that the fcc to-hcp transition occurs martensitically between 3 and 70 GPa in diamond-anvil cells without an intermediate phase. These data also reveal that the transition occurs by the introduction of stacking disorder in the fcc lattice at low pressure, which grows into hcp domains with increasing pressure. The small energy difference between the hcp and the fcc structures may allow the two phases to coexist over a wide pressure range. Evidence of similar stacking disorder and incipient growth of an hcp phase are also observed in solid Kr. PMID- 11384262 TI - Search for neutral supersymmetric Higgs bosons in pp collisions at sqrt[s] = 1.8TeV. AB - We present the results of a search for neutral Higgs bosons produced in association with b quarks in pp-->bb(phi)-->bbb final states with 91+/-7 pb(-1) of pp collisions at sqrt[s] = 1.8 TeV recorded by the Collider Detector at Fermilab. We find no evidence of such a signal and the data are interpreted in the context of the neutral Higgs sector of the minimal supersymmetric extension of the standard model. With basic parameter choices for the supersymmetric scale and the stop-quark mixing, we derive 95% C.L. lower mass limits for neutral Higgs bosons for tan(beta) values in excess of 35. PMID- 11384282 TI - Thermodynamics of C incorporation on Si(100) from ab initio calculations. AB - We study the thermodynamics of C incorporation on Si(100), a system where strain and chemical effects are both important. Our analysis is based on first principles atomistic calculations to obtain the important lowest-energy structures, and a classical effective Hamiltonian which is employed to represent the long-range strain effects and incorporate the thermodynamic aspects. We determine the equilibrium phase diagram in temperature and C chemical potential, which allows us to predict the mesoscopic structure of the system that should be observed under experimentally relevant conditions. PMID- 11384283 TI - Structure and generation mechanism of the peroxy-radical defect in amorphous silica. AB - We provide a new model of the peroxy-radical defect in amorphous silica on the basis of quantum-chemical calculations applied to clusters of atoms to model the defect. In this model, the 29Si hyperfine splittings of the peroxy radical arise from a single silicon, in agreement with the previous experimental findings. Furthermore, we show that the present model of the peroxy radical is consistent with the diffusion-limited anneal mechanism of the E'gamma center, although our model of the E'gamma center is different from the conventional charged oxygen vacancy model. PMID- 11384284 TI - Oxygen self-diffusion in alpha-quartz. AB - We have studied the formation energy of the simplest oxygen defects in alpha quartz, the oxygen vacancy and interstitial, by an ab initio approach based on density functional theory in the local density approximation. We have determined the formation energies and entropies and the migration paths and energies. From our results we can conclude that oxygen diffuses in quartz by an interstitial mechanism: the interstitial has a dumbbell structure; one of the constitutive atoms jumps towards a neighboring oxygen site. The activation energy amounts to 4.7 eV in the intrinsic regime and 2.8 eV in the extrinsic regime. PMID- 11384285 TI - High-pressure elastic properties of solid argon to 70 GPa. AB - The acoustic velocities, adiabatic elastic constants, bulk modulus, elastic anisotropy, Cauchy violation, and density in an ideal solid argon (Ar) have been determined at high pressures up to 70 GPa in a diamond anvil cell by making new approaches of Brillouin spectroscopy. These results place the first complete study for elastic properties of dense Ar and provide an improved basis for making the theoretical calculations of rare-gas solids over a wide range of compression. PMID- 11384286 TI - Peierls transition with acoustic phonons and solitwistons in carbon nanotubes. AB - We show that the Peierls instability can result in softening of acoustic phonons with small wave vectors and suggest that this unusual transition takes place in carbon nanotubes, resulting in a static twist deformation of the nanotube lattice. The topological excitations in the ordered phase are immobile and propagate only in pairs. PMID- 11384287 TI - Anisotropic domain growth of the axial next-nearest-neighbor Ising model at low temperatures. AB - We investigate the ordering kinetics for the axial next-nearest-neighbor Ising (ANNNI) model in one and two dimensions by the multispin heat bath dynamical simulation. This dynamics enables us to overcome the pinning effect and to observe the dynamical scaling law for domain growth in the ANNNI model at zero temperature. The domain growth exponent is 1/2 isotropically both in the ferromagnetic and the dry (commensurate) antiphase. In the wet (commensurate) antiphase, however, it is approximately 1/3 in the modulated direction, whereas it remains 1/2 in the nonmodulated direction. We suggest that these exponent values are dictated by 3- and 4-body diffusion-reaction processes of domain walls. PMID- 11384288 TI - Phase diagram of the A and B phases of superfluid 3He in aerogel. AB - We report the first measurements of the A-B phase transition of superfluid 3He confined within 98% silica aerogel in high magnetic fields and low temperatures. A disk of aerogel is attached to a vibrating wire resonator. The resonant frequency yields a measure of the superfluid fraction rho(s)/rho of the 3He within the aerogel. The inferred rho(s)/rho value increases substantially at the A-to- B transition of the confined superfluid, allowing us to map the A-B phase diagram as a function of field and temperature. At 4.8 bars, the B-T transition curve looks very similar to that in bulk with a simple reduction factor of order 0.45 for both transition field and temperature. PMID- 11384289 TI - Deep-inelastic neutron scattering determination of the single-particle kinetic energy in solid and liquid 3He. AB - An experimental determination of the single-particle mean kinetic energies for 3He along the T = 2.00 K isotherm in the dense liquid and in the solid hcp and bcc phases is reported. Deep-inelastic neutron scattering measurements at exchanged wave vectors ranging from 90.0 to 140.0 A(-1) have been performed in order to evaluate, within the framework of the impulse approximation, the molar volume dependence of the kinetic energy. The results are found in excellent agreement with recent diffusion Monte Carlo simulations. PMID- 11384290 TI - Diffusion kinetics in the Pd/Cu(001) surface alloy. AB - We use atom-tracking scanning tunneling microscopy to study the diffusion of Pd in the Pd/Cu(001) surface alloy as a function of temperature. By following the motion of individual Pd atoms incorporated in the surface, we show that Pd diffuses by a vacancy-exchange mechanism. We measure an activation energy for the diffusion of incorporated Pd atoms of 0.88 eV, which is in good agreement with our ab initio calculated energy of 0.94 eV. PMID- 11384291 TI - Nanoadhesion between rough surfaces. AB - A model is developed to describe the adhesion between deformable fractal surfaces over the mesoscopic realm that covers the familiar range of interest in nanotechnology from atomic dimensions to microns. This model helps us gain a quantitative understanding of the variation of adhesion with surface energy, with microstructure of rough surfaces, and with bulk deformability. The present analysis goes beyond the Gaussian distribution of asperity heights by investigating the influence of the microstructure of self-affine fractal surfaces. Our calculation reveals that orders of magnitude increase in adhesion are possible as the roughness exponent decreases. PMID- 11384292 TI - Modified critical correlations close to modulated and rough surfaces. AB - Correlation functions are sensitive to the presence of a boundary. Surface modulations give rise to modified near surface correlations, which can be measured by scattering probes. To determine these correlations, we develop a perturbative calculation in deformations in height from a flat surface. The results, combined with a renormalization group around four dimensions, are also used to predict critical behavior near a self-affine rough surface. We find that a large enough roughness exponent can modify surface critical behavior. PMID- 11384293 TI - Growth of Au clusters on amorphous Al2O3: evidence of cluster mobility above a critical size. AB - We study the 3D growth of clusters during the deposition of Au atoms on amorphous Al2O3. By comparing transmission electron microscopy images of the growth with Monte Carlo simulations, we show that nucleation takes place on substrate defects, but that further stages of growth imply that clusters leave the defects after they have reached a given critical size, and diffuse. An interesting consequence of this property is that, in contrast to intuition, and in a certain range of size, larger clusters are more mobile than smaller ones in this system. PMID- 11384294 TI - Alternating tip splitting in directional solidification. AB - We report experimental results on the tip splitting dynamics of seaweed growth in directional solidification of succinonitrile alloys. Despite the random appearance of the growth, a tip splitting morphology was observed in which the tip alternately splits to the left and to the right. The tip splitting frequency f was found to be related to the growth velocity V as a power law f~V1.5. This finding is consistent with the predictions of a tip splitting model that is also presented. Small anisotropies are shown to lead to different kinds of seaweed morphologies. PMID- 11384295 TI - Kinetic physical etching for versatile novel design of well ordered self-affine nanogrooves. AB - Ion bombardment at extreme grazing incidence leads to the formation of remarkably well ordered, one to two atomic layer deep, parallel grooves on a Cu(001) surface. These self-organized grooves are oriented parallel to the ion's plane of incidence and their period can be controlled between approximately 4 and 15 nm. We have identified two distinct temperature regimes: Between about 200 and 300 K the substrate temperature controls the period, while below 200 to about 150 K the ion energy does. The groove separation distribution shows distinct self-affine character. The physical origin of the novel nanostructures is discussed. PMID- 11384296 TI - Dynamical mean-field theory for pairing and spin gap in the attractive hubbard model. AB - We solve the attractive Hubbard model for arbitrary interaction strengths within dynamical mean-field theory. We compute the transition temperature for superconductivity and analyze electron pairing in the normal phase. The normal state is a Fermi liquid at weak coupling and a non-Fermi-liquid state with a spin gap at strong coupling. Away from half filling, the quasiparticle weight vanishes discontinuously at the transition between the two normal states. PMID- 11384297 TI - Fermi surface and magnetic structure of TmGa3. AB - We carry out measurements of the two-dimensional angular correlation of the positron annihilation radiation (2D-ACAR) to reconstruct the complex multisheet Fermi surface (FS) of the cubic rare-earth (RE) compound TmGa3. We discover a correlation between the antiferromagnetic structures and the nesting of the FS along the [110] directions. Moreover, we propose methods to estimate the density of states at the Fermi energy ( EF) and the electronic contribution to the specific heat [we obtain N(EF) = 13.6 states/Ryd cell and gamma = 2.4 mJ/mole K2]. PMID- 11384298 TI - Influence of two-pair continuum correlations following resonant excitation of excitons. AB - The memory structure induced by coherent transitions to the exciton-exciton scattering continuum is shown to have significant influence on spectrally resolved four-wave-mixing signals even under selective excitation of 1s excitons. Comparisons between experiments and calculations that account nonperturbatively for these quantum kinetic Coulomb correlations demonstrate large compensations between mean-field contributions and transitions to the two-pair continuum. Experiments with different polarizations of the laser pulses show that two-pair continuum correlations are responsible for delay-time dependent shifts of the excitonic emission as well as for substantial deformations of the line shape. PMID- 11384299 TI - Quasidegenerate self-trapping in one-dimensional charge transfer exciton. AB - The self-trapping by the nondiagonal particle-phonon interaction between two quasidegenerate energy levels of the excitonic system is studied. We propose this is realized in the charge-transfer exciton, where the directions of the polarization give the quasidegeneracy. It is shown that this mechanism, unlike the conventional diagonal one, allows a coexistence and resonance of the free and self-trapped states even in one-dimensional systems and a quantitative theory for the optical properties (light absorption and time-resolved luminescence) of the resonating states is presented. This theory gives a consistent resolution for the long-standing puzzles in quasi-one-dimensional compound A-PMDA. PMID- 11384300 TI - Partition noise and statistics in the fractional quantum hall effect. AB - A microscopic theory of current partition in fractional quantum Hall liquids, described by chiral Luttinger liquids, is developed to compute the noise correlations, using the Keldysh technique. In this Hanbury-Brown and Twiss geometry, at Laughlin filling factors nu = 1/3, the real time noise correlator exhibits oscillations which persist over larger time scales than that of an uncorrelated Hall fluid. The zero frequency noise correlations are negative at filling factor 1/3 as for bare electrons (antibunching), but are strongly reduced in amplitude. These correlations become positive (bunching) for nu < or = 1/5, suggesting a tendency towards bosonic behavior. PMID- 11384301 TI - Band splitting for Si(557)-Au: is it spin-charge separation? AB - It has been proposed that the Si(557)-Au surface exhibits spin-charge separation in a one-dimensional electron liquid. Two narrowly spaced bands are found which exhibit a well-defined splitting at the Fermi level. That is incompatible with the assignment to a spinon-holon pair in a Luttinger liquid. Instead, we propose that the two bands are associated with two nearly degenerate atomic chains, or a chain of step atoms with two broken bonds. Such an assignment explains why the surface is metallic despite an even number of electrons per unit cell. PMID- 11384302 TI - Fano resonances as a probe of phase coherence in quantum dots. AB - In the presence of direct trajectories connecting source and drain contacts, the conductance of a quantum dot may exhibit resonances of the Fano type. Since Fano resonances result from the interference of two transmission pathways, their line shape (as described by the Fano parameter q) is sensitive to dephasing in the quantum dot. We show that under certain circumstances the dephasing time can be extracted from a measurement of q for a single resonance. We also show that q fluctuates from level to level, and we calculate its probability distribution for a chaotic quantum dot. Our results are relevant to recent experiments by Gores et al. [Phys. Rev. B 62, 2188 (2000)]. PMID- 11384303 TI - Landau diamagnetism and magnetization of interacting disordered conductors. AB - We show how the orbital magnetization of an interacting diffusive electron gas can be simply related to the magnetization of the noninteracting system having the same geometry. This result is applied to the persistent current of a mesoscopic ring and to the relation between Landau diamagnetism and the interaction correction to the magnetization of diffusive systems. The field dependence of this interaction contribution can be deduced directly from the de Haas-van Alphen oscillations of the free electron gas. Known results for the free orbital magnetism of finite systems can be used to derive the interaction contribution in the diffusive regime in various geometries. PMID- 11384304 TI - Magnetization measurements of magnetic two-dimensional electron gases. AB - We directly measure the magnetization of both the conduction electrons and Mn2+ ions in (Zn,Cd,Mn)Se two-dimensional electron gases (2DEGs) by integrating them into ultrasensitive micromechanical magnetometers. The interplay between spin and orbital energy in these magnetic 2DEGs causes Landau level degeneracies at the Fermi energy. These Landau level crossings result in novel features in the de Haas-van Alphen oscillations, which are quantitatively reproduced by a simple model. PMID- 11384305 TI - Detection of single spin decoherence in a quantum dot via charge currents. AB - We consider a quantum dot attached to leads in the Coulomb blockade regime that has a spin 1 / 2 ground state. We show that, by applying an ESR field to the dot spin, the stationary current in the sequential tunneling regime exhibits a new resonance peak whose linewidth is determined by the single spin decoherence time T2. The Rabi oscillations of the dot spin are shown to induce coherent current oscillations from which T2 can be deduced in the time domain. We describe a spin inverter which can be used to pump current through a double dot via spin flips generated by ESR. PMID- 11384306 TI - Effective Lorentz force due to small-sngle impurity scattering: magnetotransport in high- Tc superconductors. AB - We show that a scattering rate which varies with angle around the Fermi surface has the same effect as a periodic Lorentz force on magnetotransport coefficients. This effect, together with the marginal Fermi liquid inelastic scattering rate, gives a quantitative explanation of the temperature dependence and the magnitude of the observed Hall effect and magnetoresistance with just the measured zero field resistivity as input. PMID- 11384307 TI - Superconductivity of metallic boron in MgB2. AB - Boron in MgB2 forms stacks of honeycomb layers with magnesium as a space filler. Band structure calculations indicate that Mg is substantially ionized, and the bands at the Fermi level derive mainly from B orbitals. Strong bonding with an ionic component and considerable metallic density of states yield a sizable electron-phonon coupling. Together with high phonon frequencies, which we estimate via zone-center frozen phonon calculations to be between 300 and 700 cm( 1), this produces a high critical temperature, consistent with recent experiments. Thus MgB2 can be viewed as an analog of the long sought, but still hypothetical, superconducting metallic hydrogen. PMID- 11384308 TI - Effects of vortex pinning and thermal fluctuations on the Josephson plasma resonance in Tl2Ba2CaCu2O8 and YBa2Cu3O6.5. AB - We investigated the temperature dependence and c-axis magnetic field dependence of the Josephson plasma resonance in optimally doped Tl2Ba2CaCu2O8 thin films and underdoped YBa2Cu3O6.5 (YBCO) ortho-II single crystals using infrared spectroscopy. We obtained the c-axis penetration depths, at low temperature, in zero fields of about 20 and 7 microm, respectively. While the temperature dependencies of the resonances in the two compounds are very similar, the magnetic field dependence in YBCO is much weaker. We attribute this weak magnetic field dependence to the lower anisotropy of YBCO and interpret the observed behaviors in terms of thermal fluctuations and pinning of pancake vortices. PMID- 11384310 TI - Interacting random Dirac fermions in superconducting cuprates. AB - We study the effects of quasiparticle interactions on disorder-induced localization of Dirac-like nodal excitations in superconducting high- Tc cuprates. As suggested by the experimental angle-resolved photoemission spectroscopy and terahertz conductivity data in Bi2Sr2CaCu2O(8+delta), we focus on the interactions mediated by the order parameter fluctuations near an incipient second pairing transition d --> d + is(id'). We find interaction corrections to the density of states, specific heat, and conductivity as well as phase and energy relaxation rates and assess the applicability of the recent localization scenarios for noninteracting random Dirac fermions to the cuprates. PMID- 11384309 TI - Unique spin dynamics and unconventional superconductivity in the layered heavy fermion compound CeIrIn5: NQR evidence. AB - We report measurements of the 115In nuclear spin-lattice relaxation rate ( 1/T1) between T = 0.09 and 100 K in the new heavy fermion (HF) compound CeIrIn5. At 0.4 < or = T< or = 100 K, 1/T1 is strongly T-dependent, which indicates that CeIrIn5 is much more itinerant than known Ce-based HFs. We find that 1/T1T, subtracting that for LaIrIn5, follows a (1 / T+straight theta)3/4 variation with straight theta = 8 K. We argue that this novel feature points to anisotropic, due to a layered crystal structure, spin fluctuations near a magnetic ordering. The bulk superconductivity sets in at 0.40 K below which the coherence peak is absent and 1/T1 follows a T3 variation, which suggests unconventional superconductivity with line-node gap. PMID- 11384311 TI - Incipient nodal pairing in planar d-wave superconductors. AB - The possibility of a second pairing transition d --> d + is ( d + id') in planar d-wave superconductors which occurs in the absence of external magnetic field, magnetic impurities, or boundaries is established in the framework of the nonperturbative phenomenon of dynamical chiral symmetry breaking in the system of (2+1)-dimensional Dirac-like nodal quasiparticles. We determine the critical exponents and quasiparticle spectral functions that characterize the corresponding quantum-critical behavior and discuss some of its potentially observable spectral and transport features. PMID- 11384312 TI - Magnetic anisotropy of a single cobalt nanocluster. AB - Using a new micro-SQUID setup, we investigate magnetic anisotropy in a single 1000-atom cobalt cluster. This system opens new fields in the characterization and understanding of the origin of magnetic anisotropy in such nanoparticles. For this purpose, we report three-dimensional switching field measurements performed on a 3 nm cobalt cluster embedded in a niobium matrix. We are able to separate the different magnetic anisotropy contributions and evidence the dominating role of the cluster surface. PMID- 11384313 TI - Role of dynamic Jahn-Teller distortions in Na2C60 and Na2CsC60 studied by NMR. AB - Through 13C NMR spin lattice relaxation ( T1) measurements in cubic Na2C60, we detect a gap in its electronic excitations, similar to that observed in tetragonal A4C60. This establishes that Jahn-Teller distortions (JTD) and strong electronic correlations must be considered to understand the behavior of even electron systems, regardless of the structure. Furthermore, in metallic Na2CsC60, a similar contribution to T1 is also detected for 13C and 133Cs NMR, implying the occurrence of excitations typical of JT distorted C( 2-)60 (or equivalently C( 4 )60). This supports the idea that dynamic JTD can induce attractive electronic interactions in odd electron systems. PMID- 11384314 TI - Subthreshold carrier-LO phonon dynamics in semiconductors with intermediate polaron coupling: a purely quantum kinetic relaxation channel. AB - Femtosecond transmission spectra of highly polar CdTe are compared to more covalent GaAs contrasting semiclassical kinetics with two-time quantum kinetics based on the Dyson equation. Nonequilibrium heavy holes in CdTe show ultrafast energy redistribution via the Frohlich mechanism even if photoexcited below the LO phonon energy. This subthreshold relaxation is a genuine quantum kinetic effect. It gains importance if the polaron self-energy is comparable to the phonon energy. Conservation of the free-particle energies is not required under these conditions. PMID- 11384315 TI - Controlling the interaction between light and gold nanoparticles: selective suppression of extinction. AB - The interaction of visible light with the particle-plasmon resonance of metallic nanoparticles can be controlled by geometrical arrangement of nanoparticle arrays. These arrays are placed on a substrate that supports guided modes in the wavelength range of the particle plasmon. Coupling of this particle-plasmon resonance to the directly incident light and to the waveguide modes results in almost complete suppression of light extinction within narrow spectral bands due to destructive interference. Variation of the structure parameters allows continuous tuning of these high-transmission bands across the particle-plasmon resonance. PMID- 11384316 TI - Origin of negative thermal expansion in cubic ZrW2O8 revealed by high pressure inelastic neutron scattering. AB - Isotropic negative thermal expansion has been reported in cubic ZrW2O8 over a wide range of temperatures (0-1050 K). Here we report the direct experimental determination of the Gruneisen parameters of phonon modes as a function of their energy, averaged over the whole Brillouin zone, by means of high pressure inelastic neutron scattering measurements. We observe a pronounced softening of the phonon spectrum at P = 1.7 kbar compared to that at ambient pressure by about 0.1-0.2 meV for phonons of energy below 8 meV. This unusual phonon softening on compression, corresponding to large negative Gruneisen parameters, is able to account for the observed large negative thermal expansion. PMID- 11384318 TI - Bubble dynamics relaxation in aqueous foam probed by multispeckle diffusing-wave spectroscopy. AB - We study the bubble rearrangement dynamics in aqueous foam during the passage from liquidlike to solidlike behavior which follows a transient shear deformation that perturbs the bubble packing. The local dynamics is probed using multispeckle diffusing-wave spectroscopy. We show that following the perturbation the average time between rearrangements relaxes exponentially, with time elapsed since the end of the perturbation. The observed scaling of the characteristic relaxation time with perturbation amplitude and foam age is explained by a schematic coarse grained model based on the scaling state hypothesis. PMID- 11384317 TI - Phase separation in mixtures of colloids and long ideal polymer coils. AB - Colloidal suspensions with free polymer coils which are larger than the colloidal particles are considered. The polymer-colloid interaction is modeled by an extension of the Asakura-Oosawa model. Phase separation occurs into dilute and dense fluid phases of colloidal particles when polymer is added. The critical density of this transition tends to zero as the size of the polymer coils diverges. PMID- 11384319 TI - Dynamics of coarsening foams: accelerated and self-limiting drainage. AB - The evolution of a foam is determined by drainage flow of the continuous (liquid) phase and coarsening (aging) of the dispersed phase (gas bubbles). Free-drainage experiments with slow- and fast-coarsening gases show markedly different dynamics and elucidate the importance of the coupling of the two effects. Strong coarsening leads to drainage times that are shorter (accelerated drainage) and independent of the initial liquid content (self-limiting drainage). A model incorporating the physics of both drainage and diffusive coarsening shows quantitative agreement with experiment. PMID- 11384320 TI - Comment on "low-dimensional Bose liquids: beyond the Gross-Pitaevskii approximation". PMID- 11384322 TI - Comment on "dispersion-independent high-visibility quantum interference in ultrafast parametric down-conversion". PMID- 11384325 TI - Comment on "atomic transport and chemical stability during annealing of ultrathin Al2O3 films on Si". PMID- 11384323 TI - Comment on "radial-fluctuation-induced stabilization of the ordered state in two dimensional classical clusters". PMID- 11384327 TI - Comment on "analytic structure of one-dimensional localization theory: reexamining Mott's law". PMID- 11384328 TI - Comment on "two time scales and violation of the fluctuation-dissipation theorem in a finite dimensional model for structural glasses". PMID- 11384330 TI - Interplay of genetic and actual lifespans. PMID- 11384332 TI - Measuring the quantum state of a large angular momentum. AB - We demonstrate a general method to measure the quantum state of an angular momentum of arbitrary magnitude. The (2F+1)x(2F+1) density matrix is completely determined from a set of Stern-Gerlach measurements with (4F+1) different orientations of the quantization axis. We implement the protocol for laser cooled Cesium atoms in the 6S(1/2)(F = 4) hyperfine ground state and apply it to a variety of test states prepared by optical pumping and Larmor precession. A comparison of input and measured states shows typical reconstruction fidelities F >/~0.95. PMID- 11384333 TI - Superfluid gyroscope with cold atomic gases. AB - A trapped Bose-Einstein condensed atomic gas containing a quantized vortex is predicted to exhibit precession after a sudden rotation of the confining potential. The equations describing the motion of the condensate are derived and the effects of superfluidity explicitly pointed out. The dependence of the precession frequency on the relevant parameters of the problem is discussed. The proposed gyroscope is well suited to explore rotational effects at the level of single quanta of circulation. PMID- 11384334 TI - Quasicontinuous atom laser in the presence of gravity. AB - We analyze the extraction from a trapped Bose-Einstein condensate to a nontrapping state of a coherent atomic beam and its subsequent fall, at T = 0 K. Our treatment fully takes gravity into account but neglects the mean-field potential exerted on the free falling beam by the trapped atoms. We derive analytical expressions for the output rate and the output mode of the quasicontinuous "atom laser." Comparison with experimental data of Bloch et al. [Phys. Rev. Lett. 82, 3008 (1999)] is satisfactory. PMID- 11384335 TI - Uniting Bose-Einstein condensates in optical resonators. AB - The relative phase of two initially independent Bose-Einstein condensates can be laser cooled to unite the two condensates by putting them into a ring cavity and coupling them with an internal Josephson junction. First, we show that this phase cooling process already appears within a semiclassical model. We calculate the stationary states, find regions of bistable behavior, and suggest a Ramsey-type experiment to measure the buildup of phase coherence between the condensates. We also study quantum effects and imperfections of the system. PMID- 11384336 TI - Coherence resonance in coupled chaotic oscillators. AB - Existing works on coherence resonance, i.e., the phenomenon of noise-enhanced temporal regularity, focus on excitable dynamical systems such as those described by the FitzHugh-Nagumo equations. We extend the scope of coherence resonance to an important class of dynamical systems: coupled chaotic oscillators. In particular, we show that, when a system of coupled chaotic oscillators is under the influence of noise, the degree of temporal regularity of dynamical variables characterizing the difference among the oscillators can increase and reach a maximum value at some optimal noise level. We present numerical results illustrating the phenomenon and give a physical theory to explain it. PMID- 11384337 TI - Solving N-body problems with neural networks. AB - We show a new approach for solving the N-body problems based on neural networks. Without loss of generality, we derived a network solution for the time-dependent positions of N bodies in self-gravitating systems. The simulation is limited to a system of collisionless disks-a case for determining the spatial distributions of dark matter and in reproducing global effects such as formation of spiral galaxies. Our approach yields a solution that is analytic with time-reversed path tracing capabilities that could lead to new findings in the study of the collective behavior of N-body systems. PMID- 11384338 TI - Plasmoid impacts on neutron stars and highest energy cosmic rays. AB - Particle acceleration by electrostatic polarization fields that arise in plasmas streaming across magnetic fields is discussed as a possible acceleration mechanism of highest energy ( greater, similar10(20) eV) cosmic rays. Specifically, plasmoids arising in planetoid impacts onto neutron star magnetospheres are considered. We find that such impacts at plausible rates may account for the observed flux and energy spectrum of the highest energy cosmic rays. PMID- 11384339 TI - E10, BE10 and arithmetical chaos in superstring cosmology. AB - It is shown that the neverending oscillatory behavior of the generic solution, near a cosmological singularity, of the massless bosonic sector of superstring theory can be described as a billiard motion within a simplex in nine-dimensional hyperbolic space. The Coxeter group of reflections of this billiard is discrete and is the Weyl group of the hyperbolic Kac-Moody algebra E10 (for type II) or BE10 (for type I or heterotic), which are both arithmetic. These results lead to a proof of the chaotic ("Anosov") nature of the classical cosmological oscillations, and suggest a "chaotic quantum billiard" scenario of vacuum selection in string theory. PMID- 11384340 TI - Beta function for anisotropic current interactions in 2D. AB - By making use of current-algebra Ward identities we study renormalization of general anisotropic current-current interactions in 2D. In this prescription we propose a compact expression for the beta function to all orders. PMID- 11384341 TI - (De)constructing dimensions. AB - We construct renormalizable, asymptotically free, four-dimensional gauge theories that dynamically generate a fifth dimension. PMID- 11384342 TI - On some positivity properties of the interquark potential in QCD. AB - We argue that the Fourier transform of the exponential e(-betaV(R)) of the static interquark potential in QCD is positive. It has been shown by Lieb some time ago that this property allows in the same limit of static, spin independent potential, to prove a certain mass relation between baryons with different quark flavors. PMID- 11384343 TI - In-medium properties of the D13(1520) nucleon resonance. AB - The in-medium properties of the D13(1520) nucleon resonance were studied via photoproduction of pi(0) mesons from nuclei (C, Ca, Nb, Pb) with the TAPS detector at the Mainz Microton accelerator. The inclusive (single and multiple pion production) data disagree with model predictions which explain the disappearance of the second resonance bump in total photoabsorption via a medium modification of the D13-->Nrho decay. The exclusive single pi(0) production data show no broadening of the resonance structure beyond Fermi smearing. Both results together cast doubt on attempts to explain the vanishing of the second resonance bump for nuclei by a broadening of the D13 resonance. PMID- 11384345 TI - Observation of color-transparency in diffractive dissociation of pions. AB - We have studied the diffractive dissociation into dijets of 500 GeV/c pions scattering coherently from carbon and platinum targets. Extrapolating to asymptotically high energies (where t(min)-->0), we find that when the per nucleus cross section for this process is parametrized as sigma = sigma0Aalpha, alpha has values near 1.6, the exact result depending on jet transverse momentum. These values are in agreement with those predicted by theoretical calculations of color-transparency. PMID- 11384344 TI - Direct measurement of the pion valence-quark momentum distribution, the pion light-cone wave function squared. AB - We present the first direct measurements of the pion valence-quark momentum distribution which is related to the square of the pion light-cone wave function. The measurements were carried out using data on diffractive dissociation of 500 GeV/c pi(-) into dijets from a platinum target at Fermilab experiment E791. The results show that the /q&q> light-cone asymptotic wave function describes the data well for Q2 approximately 10 (GeV/c)(2) or more. We also measured the transverse momentum distribution of the diffractive dijets. PMID- 11384347 TI - Flow at the SPS and RHIC as a quark-gluon plasma signature. AB - Radial and elliptic flow in noncentral heavy-ion collisions can constrain the effective equation of state (EOS) of the excited nuclear matter. To this end, a model combining relativistic hydrodynamics and a hadronic transport code [Sorge, Phys. Rev. C 52, 3291 (1995)] is developed. For an EOS with a first-order phase transition, the model reproduces both the radial and elliptic flow data at the SPS. With the EOS fixed from SPS data, we quantify predictions at RHIC where the quark-gluon plasma (QGP) pressure is expected to drive additional radial and elliptic flows. Currently, the strong elliptic flow observed in the first RHIC measurements does not conclusively signal this nascent QGP pressure. PMID- 11384348 TI - Three- and four-nucleon systems from chiral effective field theory. AB - Recently developed chiral nucleon-nucleon (NN) forces at next-to-leading order (NLO), that describe NN phase shifts up to about 100 MeV fairly well, have been applied to 3N and 4N systems. Faddeev-Yakubovsky equations have been solved rigorously. The resulting 3N and 4N binding energies are in the same range as found using standard NN potentials. In addition, low-energy 3N scattering observables are very well reproduced as for standard NN forces. The long-standing A(y) puzzle is absent at NLO. The cutoff dependence of the scattering observables is rather weak. PMID- 11384349 TI - New mechanism for the enhancement of sd dominance in interacting boson models. AB - We introduce an exactly solvable model for interacting bosons that extend up to high spin and interact through a repulsive pairing force. The model exhibits a phase transition to a state with almost complete sd dominance. The repulsive pairing interaction that underlies the model has a natural microscopic origin in the Pauli exclusion principle between constituent nucleons. As such, repulsive pairing between bosons seems to provide a new mechanism for the enhancement of sd dominance, giving further support for the validity of the sd interacting boson model. PMID- 11384350 TI - Sharp spectral lines observed in gamma-ray ionized parahydrogen crystals. AB - We have observed sharp infrared spectral lines ( Deltanu approximately 60 MHz FWHM) in gamma-ray ionized para- H2 crystals. The lines are assigned to the Q1(0) transition of H2 near H+3 and H-, which becomes optically active and Stark shifted by the Coulomb fields of the ions. A simple model calculation gives a spectrum which agrees approximately with the observation. The spectrum stays the same over many days, demonstrating the stability of the ionized system. The remarkable sharpness and reproducibility of the line positions indicate uniformity of the local structure surrounding the charges. PMID- 11384351 TI - Motional effect in surface sum-frequency vibrational spectroscopy. AB - Fast orientational motion of molecules at a surface can affect the sum-frequency vibrational spectra of the surface. Calculation shows that the effect is significant if the molecular orientation varies over a broad range within the vibrational relaxation time. The stretch vibration of the free OH bonds at the vapor/water interface is used to illustrate the importance of the effect. PMID- 11384352 TI - Charge transfer and elastic scattering in very slow H(+)+D(1s) half collisions. AB - Charge transfer and elastic scattering probabilities were measured for half collisions between very slow protons and atomic deuterium. Collision energies down to a few meV, lower by more than an order of magnitude and with better energy resolution than previous measurements, were studied using the dissociation of the HD+ electronic ground state. The collision energy is determined a posteriori from the measured momentum vector of the dissociating charged fragments. The experimental results are in good agreement with our coupled channel calculations. PMID- 11384346 TI - Midrapidity antiproton-to-proton ratio from Au+Au collisions at sqrt [s(NN)]=130 GeV. AB - We report results on the ratio of midrapidity antiproton-to-proton yields in Au+Au collisions at sqrt[s(NN)] = 130 GeV per nucleon pair as measured by the STAR experiment at RHIC. Within the rapidity and transverse momentum range of /y/<0.5 and 0.41 an "ordered" state with straight phi not equal0 occurs at h = 0, representing an extreme breakdown of equipartition. Critical slowing and qualitative changes in the velocity distribution function near the transition are noted. PMID- 11384358 TI - Passive scalar intermittency in low temperature helium flows. AB - We report new measurements of mixing of passive temperature field in a turbulent flow. The use of low temperature helium gas allows us to span a range of microscale Reynolds number, R(lambda), from 100 to 650. The exponents xi(n) of the temperature structure functions approximately r(xi(n)) are shown to saturate to xi(infinity) approximately 1.45+/ 0.1 for the highest orders, n approximately 10. This saturation is a signature of statistics dominated by frontlike structures, the cliffs. Statistics of the cliffs' characteristics are performed, particularly their widths are shown to scale as the Kolmogorov length scale. PMID- 11384359 TI - Anisotropic homogeneous turbulence: hierarchy and intermittency of scaling exponents in the anisotropic sectors. AB - We present the first measurements of anisotropic statistical fluctuations in perfectly homogeneous turbulent flows. We address both problems of intermittency in anisotropic sectors and hierarchical ordering of anisotropies on a direct numerical simulation of a three dimensional random Kolmogorov flow. We achieved an homogeneous and anisotropic statistical ensemble by randomly shifting the forcing phases. We observe high intermittency as a function of the order of the velocity correlation within each fixed anisotropic sector and a hierarchical organization of scaling exponents at fixed order of the velocity correlation at changing the anisotropic sector. PMID- 11384360 TI - Signature of turbulent zonal flows observed in the DIII-D tokamak. AB - The spectrum of turbulent density fluctuations at long poloidal wavelengths in the edge plasma of the DIII-D tokamak peaks at nonzero radial wave number. The associated electric-potential fluctuations cause sheared E x B flows primarily in the poloidal direction. These zonal flows have been predicted by theory and are believed to regulate the overall level of turbulence and anomalous transport. This study provides the first indirect experimental identification of zonal flows. PMID- 11384361 TI - Liquid alumina: detailed atomic coordination determined from neutron diffraction data using empirical potential structure refinement. AB - The neutron scattering structure factor S(N)(Q) for a 40 mg drop of molten alumina (Al2O3) held at 2500 K, using a laser-heated aerodynamic levitation furnace, is measured for the first time. A 1700 atom model of liquid alumina is generated from these data using the technique of empirical potential structural refinement. About 62% of the aluminum sites are 4-fold coordinated, matching the mostly triply coordinated oxygen sites, but some 24% of the aluminum sites are 5 fold coordinated. The octahedral aluminum sites found in crystalline alpha-Al2O3 occur only at the 2% level in liquid alumina. PMID- 11384362 TI - Effect of a rippling mode on resonances of carbon nanotubes. AB - A recent study determined the Young's modulus of carbon nanotubes by measuring resonance frequency and using the modulus-frequency relation resulting from the linear vibration theory. It leads to the report that the Young's modulus decreases sharply, from about 1 to 0.1 TPa with the diameter D increasing from 8 to 40 nanometers, and the investigators attributed this decrease to the emergence of an unusual bending mode during the measurement that corresponds to rippling on the inner arc of the bent nanotubes. The nonlinear analysis presented in this paper that captures the rippling mode suggests that the effective Young's modulus can indeed decrease substantially with increasing diameter, and that the results from the classical linear theory may be invalid in such measurements. PMID- 11384363 TI - Soft acoustic modes in the two-dimensional spin system SrCu2(BO3)(2). AB - SrCu2(BO3)(2) is a two-dimensional dimerized quantum spin system which is close to a quantum critical point. The sound velocity for the longitudinal and transverse acoustic modes shows strong spin-lattice effects. The shear c(66) mode exhibits a pronounced softening of 4.5% as a function of temperature and softens more than 25% in fields up to 50 T. This huge effect occurs in the vicinity of the magnetization plateaus m/m(0) = 1/4 and 1/3. We can analyze quantitatively the temperature dependence of all measured elastic modes c(11), c(44), and c(66) with an exchange striction mechanism. The soft c(66) mode with B(2g) symmetry enables us to predict the possible symmetry of the condensed triplets in some plateaus. PMID- 11384364 TI - Pressure-induced Invar effect in Fe-Ni alloys. AB - We have measured the pressure-volume (P-V) relations for cubic iron-nickel alloys for three different compositions: Fe 0.64Ni (0.36), Fe 0.55Ni (0.45), and Fe 0.20Ni (0.80). It is observed that for a certain pressure range the bulk modulus does not change or can even decrease to some minimum value, after which it begins to increase under still higher pressure. In our experiment, we observe for the first time a new effect, namely, that the Fe-Ni alloys with high Ni concentrations, which show positive thermal expansion at ambient pressure, become Invar system upon compression over a certain pressure range. PMID- 11384365 TI - Particle excursions in colloidal crystals. AB - We have performed synchrotron small-angle x-ray diffraction experiments on charge stabilized colloidal crystals. The mean squared displacements, obtained from Debye-Waller factors, decrease with increasing density beyond freezing. Irrespective of the range of the repulsion, our data and previous results lie on a common curve, linearly scaled between the melting and close-packed densities. Surprisingly, the excursions are smaller than predicted by theory. PMID- 11384366 TI - Freezing of dynamical exponents in low dimensional random media. AB - A particle in a random potential with logarithmic correlations in dimensions d = 1,2 is shown to undergo a dynamical transition at T(dyn)>0. In d = 1 exact results show T(dyn) = T(c), the static glass transition temperature, and that the dynamical exponent changes from z(T) = 2+2(T(c)/T)(2) at high T to z(T) = 4T(c)/T in the glass phase. The same formulas are argued to hold in d = 2. Dynamical freezing is also predicted in the 2D random gauge XY model and related systems. In d = 1 a mapping between dynamics and statics is unveiled and freezing involves barriers as well as valleys. Anomalous scaling occurs in the creep dynamics, relevant to dislocation motion experiments. PMID- 11384367 TI - Self-similarity and pattern selection in the roughening of binary liquid films. AB - Films of spinodally decomposing binary liquid mixtures show transient wetting of both confining interfaces by one of the phases, and rupture, with characteristic wavelength lambda(c) and time tau(rupture), leading to flat droplets of the nonwetting phase encapsulated by the wetting phase. Over the entire range of film thicknesses d approximately 100-3500 nm, we find tau(rupture)~d(1.01+/-0.08), indicating film structures that scale self-similarly with d, and find also that lambda(c) approximately 60d(0.97+/-0.03), the large prefactor suggesting a rupture wavelength which minimizes the interfacial tension of the roughened film. PMID- 11384368 TI - Observation of a metastable periodic structure for the (001) surface of KTaO3 after cleaving in situ. AB - Helium atom diffraction experiments carried out under ultrahigh vacuum conditions on a freshly cleaved (001) surface of KTaO3 reveal metastable features which decay over a period of several hours. The initial He diffraction pattern contains large scattering intensity satellite peaks very close to the specular reflection beam. As time from cleaving elapses, the satellite intensities diminish virtually to zero while the specular intensity increases, and the diffraction pattern evolves into one consistent with the (1x1) bulk termination surface. The data are compared with model calculations for scattering from a series of terraces at two heights with a distribution of terrace lengths [Surf. Sci. 384, 15 (1997)]. PMID- 11384369 TI - Phase coexistence during surface phase transitions. AB - In contrast to standard thermodynamic models, we observe phase coexistence over an extended temperature range at a first-order surface phase transition. We have measured the domain evolution of the Si(111)-( 7x7) to ( 1x1) phase transition with temperature, using low-energy electron microscopy. Comparison with detailed, quantitative theoretical predictions shows that coexistence is due to long-range elastic and electrostatic domain interactions. Phase coexistence is predicted to be a universal feature of surface phase transitions. PMID- 11384370 TI - Novel layered superstructures in mixed ultralong n-alkanes. AB - Two new types of layered structures were found in binary mixtures of n-alkanes ranging from C122H246 to C294H590. At high temperatures a semicrystalline form is the stable phase, having a regular structure of alternating crystalline and amorphous layers. The two long-chain compounds are mixed in the crystalline layers and the amorphous layers consist of the surplus length of the longer chains. At lower temperatures a reversible transition occurs to a triple layer superlattice structure with a periodicity of up to 50 nm. These two new phases allow the existence of binary solid solutions of chains with a length ratio of up to 1.7 and a chain length difference of 100 CH2 groups. PMID- 11384371 TI - Epitaxial growth of a low-density framework form of crystalline silicon: a molecular-dynamics study. AB - Crystal growth processes of low-density framework forms of crystalline silicon, named Si clathrates ( Si34 and Si46), during solid phase epitaxy (SPE) have been successfully observed in molecular-dynamics simulations using the Tersoff potential. The activation energy of SPE for Si34 has been found to correspond with the experimental value ( approximately 2.7 eV) for the cubic diamond phase, while the SPE rates of Si46 are much lower than that of Si34. The structural transition from Si46 to Si34 can be also observed during the Si46-[001] SPE. The present results suggest that new wide-gap Si semiconductors with clathrate structures can be prepared using epitaxial growth techniques. PMID- 11384372 TI - Fluorescence intermittency in self-assembled InP quantum dots. AB - Fluorescence intermittency in InP self-assembled dots is investigated by means of far field imaging and single dot spectroscopy. Based on our observation that blinking dots are found in the vicinity of scratches and the blinking frequency is drastically enhanced under a near-infrared laser irradiation, we attribute the origin of the fluorescence intermittency to a local electric field due to a carrier trapped at a deep localized center in the Ga0.5In0.5P matrix. The validity of this explanation is confirmed by a thermal activation-type behavior of the switching rate and artificial reproduction of the blinking phenomenon by an external electric field. PMID- 11384373 TI - Coulomb interaction and quantum transport through a coherent scatterer. AB - An interplay between charge discreteness, coherent scattering, and Coulomb interaction yields nontrivial effects in quantum transport. We derive a real-time effective action and an equivalent quantum Langevin equation for an arbitrary coherent scatterer and evaluate its current-voltage characteristics in the presence of interactions. Within our model, at large conductances G0 and low T (but outside the instanton-dominated regime), the interaction correction to G0 saturates and causes conductance suppression by a universal factor which depends only on the type of the conductor. PMID- 11384374 TI - Surface soft phonon and the sqrt[3] x sqrt[3] <--> 3x3 phase transition in Sn/Ge(111) and Sn/Si(111). AB - Density functional theory calculations show that the reversible Sn/Ge(111) sqrt[3]xsqrt[3]<-->3x3 phase transition can be described in terms of a surface soft phonon. The isovalent Sn/Si(111) case does not display this transition since the sqrt[3]xsqrt[3] phase is the stable structure at low temperature, although it presents a partial softening of the 3x3 surface phonon. The rather flat energy surfaces for the atomic motion associated with this phonon mode in both cases explain the experimental similarities found at room temperature between these systems. The driving force underlying the sqrt[3]xsqrt[3]<-->3x3 phase transition is shown to be associated with the electronic energy gain due to the Sn dangling bond rehybridization. PMID- 11384375 TI - Fermi-liquid behavior of the low-density 2D hole gas in a GaAs/AlGaAs heterostructure at large values of r(s). AB - We examine the validity of the Fermi-liquid description of the dilute 2D hole gas in the crossover from "metallic"-to-"insulating" behavior of rho(T). It has been established that, at r(s) as large as 29, negative magnetoresistance does exist and is well described by weak localization theory. The dephasing time, extracted from the magnetoresistance, is dominated by the T2 term due to hole-hole scattering in the clean limit. The effect of hole-hole interactions, however, is suppressed when compared with the theory derived for small r(s). PMID- 11384376 TI - Isotope effects in underdoped cuprate superconductors: a quantum critical phenomenon. AB - We show that the unusual doping dependence of the isotope effects on transition temperature and zero temperature in-plane penetration depth naturally follows from the doping driven 3D-2D crossover and the 2D quantum superconductor to insulator transition in the underdoped limit. Since lattice distortions are the primary consequence of isotope substitution, our analysis clearly reveals the strong involvement of lattice degrees of freedom in mediating superconductivity. PMID- 11384377 TI - Theory of equilibrium flux lattice in UPt3 under magnetic field parallel to hexagonal crystal axis. AB - We investigate Abrikosov lattice structures in the unconventional superconductor UPt (3) under magnetic field parallel to the hexagonal crystal axis. Only the two dimensional E2 superconducting state among the many other states of different symmetry is compatible with the recent observation [A. Huxley et al., Nature (London) 406, 160 (2000)] of the flux lattice in the A phase misaligned with crystallographic directions. It is shown that the inequality of the London penetration depths in the basal plane directions resulting from the superposition of hexagonal crystal and superconducting state anisotropies leads for E2 to a slightly distorted triangular flux lattice. PMID- 11384378 TI - Transport anomalies and the role of pseudogap in the 60-K phase of YBa(2)Cu(3)O(7 delta). AB - We report the result of our accurate measurements of the a- and b-axis resistivity, Hall coefficient, and the a-axis thermopower in untwinned YBa(2)Cu(3)O(y) single crystals in a wide range of doping. It is found that both the a-axis resistivity and the Hall conductivity show anomalous dependences on the oxygen content y in the 60-K phase below the pseudogap temperature T(*). The complete data set enables us to narrow down the possible pictures of the 60-K phase, with which we discuss a peculiar role of the pseudogap in the charge transport. PMID- 11384379 TI - Scanning tunneling spectroscopy of Bi(2)Sr(2)CuO(6+delta): new evidence for the common origin of the pseudogap and superconductivity. AB - Using scanning tunneling spectroscopy, we investigated the temperature dependence of the quasiparticle density of states of overdoped Bi(2)Sr(2)CuO(6+delta) between 275 mK and 82 K. Below T(c) = 10 K, the spectra show a gap with well defined coherence peaks at +/-Delta(p) approximately 12 meV, which disappear at T(c). Above T(c), the spectra display a clear pseudogap of the same magnitude, gradually filling up and vanishing at T(*) approximately 68 K. The comparison with Bi(2)Sr(2)CaCu(2)O(8+delta) demonstrates that the pseudogap and the superconducting gap scale with each other, providing strong evidence that they have a common origin. PMID- 11384380 TI - High-resolution photoemission study of MgB2. AB - We have performed high-resolution photoemission spectroscopy on MgB2 and observed opening of a superconducting gap with a narrow coherent peak. We found that the superconducting gap is s like with the gap value ( Delta) of 4.5+/-0.3 meV at 15 K. The temperature dependence (15-40 K) of the gap value follows well the BCS form, suggesting that 2Delta/k(B)T(c) at T = 0 is about 3. No pseudogap behavior is observed in the normal state. The present results strongly suggest that MgB2 is categorized into a phonon-mediated BCS superconductor in the weak-coupling regime. PMID- 11384381 TI - Reversal of momentum relaxation. AB - A new phenomenon of momentum relaxation reversal has been discovered experimentally and explained theoretically for dipolar spin waves in magnetic garnet films. It is shown that the process of momentum relaxation, caused by the scattering of a signal wave on defects, can be reversed, and the signal can be restituted after it left the scattering region. The reversal of momentum relaxation is achieved by frequency-selective parametric amplification of a narrow band of scattered waves having low group velocities and frequencies close to the frequency of the original signal wave. The phenomenon can be used for the development of a new type of active microwave delay lines. PMID- 11384382 TI - Stripes induced by orbital ordering in layered manganites. AB - Spin-charge-orbital ordered structures in doped layered manganites are investigated using an orbital-degenerate double-exchange model tightly coupled to Jahn-Teller distortions. In the ferromagnetic phase, unexpected diagonal stripes at x = 1/m ( m = integer) are observed, as in recent experiments. These stripes are induced by the orbital degree of freedom, which forms a staggered pattern in the background. A pi shift in the orbital order across stripes is identified, analogous to the pi shift in spin order across stripes in cuprates. At x = 1/4 and 1/3, another nonmagnetic phase with diagonal static charge stripes is stabilized at intermediate values of the t(2g)-spins exchange coupling. PMID- 11384383 TI - Chaotic domain patterns in periodic inhomogeneous magnetic films. AB - A study of domain structures in thin inhomogeneous ferromagnetic films is presented. It is shown that smooth and small inhomogeneities in the exchange and anisotropy parameters yield very complex domain structures. We show that the domain walls are fixed near certain inhomogeneities but do not repeat their space distribution. We found that there are metastable chaotic domain patterns in periodic inhomogeneous films. These results are relevant for magnetoresistive devices. PMID- 11384384 TI - Observation of phonon bottleneck in quantum dot electronic relaxation. AB - Time-resolved differential transmission measurements of self-assembled In0.4Ga0.6As quantum dots clearly indicate a phonon bottleneck between the n = 2 and n = 1 electronic levels. The key to this observation is the generation of electrons in dots where there are no holes so that electron-hole scattering does not mask the bottleneck. We use a simple carrier capture model consisting of two capture configurations to explain the bottleneck signal and offer arguments to rule out other possible sources of the signal. PMID- 11384385 TI - Spectrum of luminescence from laser-created bubbles in water. AB - The spectrum of the luminescence emitted at the collapse of single laser-induced bubbles in water is measured for different maximum bubble radii. Bubbles as large as 2 mm show a molecular OH(*) band at 310 nm in the spectrum, which otherwise can be fitted approximately with a blackbody curve at a temperature of 7800 K. This finding provides a connection between the light emission of single bubbles and multibubble sonoluminescence, since in the latter case the same molecular band is observed. Surface instabilities are observed in the larger bubbles, and may be connected with the OH(*) emission. PMID- 11384386 TI - Optimal cloning of coherent states with a linear amplifier and beam splitters. AB - A transformation achieving the optimal symmetric N-->M cloning of coherent states is presented. Its implementation requires only a phase-insensitive linear amplifier and a network of beam splitters. An experimental demonstration of this continuous-variable cloner should therefore be in the scope of current technology. The link between optimal quantum cloning and optimal amplification of quantum states is also pointed out. PMID- 11384387 TI - Optical implementation of continuous-variable quantum cloning machines. AB - We propose an optical implementation of the Gaussian continuous-variable quantum cloning machines. We construct a symmetric N-->M cloner which optimally clones coherent states, and we also provide an explicit design of a class of asymmetric 1-->2 cloning machines. All proposed cloning devices can be built from just a single nondegenerate optical parametric amplifier and several beam splitters. PMID- 11384388 TI - Heavy-atom skeleton quantization and proton tunneling in "intermediate-barrier" hydrogen bonds. AB - Quantum effects on proton transfer through barriers of several kcal/mol in hydrogen bonds are investigated theoretically in malonaldehyde. Such "intermediate-barrier" proton transfer processes play a key role in the catalytic activity of some enzymes. Tunneling is shown to be significant in this reaction even at room temperature. More importantly, the quantum nature of the heavy molecular frame atoms is found to substantially enhance proton tunneling. These findings have far-reaching implications for common modeling strategies of proton transfer in complex systems such as biomolecules. PMID- 11384389 TI - Optical analogy to electronic quantum corrals. AB - We describe full multiple-scattering calculations of localized surface photonic states set up by lithographically designed nanostructures made of a finite number of dielectric pads deposited on a planar surface. The method is based on a numerical solution of the dyadic Dyson's equation. When the pads are arranged to form a closed circle, we find field patterns that look like the electronic charge density recently observed above quantum corrals. We propose two experimental techniques that could be used to observe these electromagnetic modes in direct space. PMID- 11384390 TI - Clustering of arrays of chaotic chemical oscillators by feedback and forcing. AB - Feedback and external forcing are applied to an array of chaotic electrochemical oscillators through variations in the applied potential. We see transitions from intermittent clusters to stable chaotic clusters to stable periodic clusters to synchronized states as the feedback gain and forcing amplitude, respectively, are varied. With forcing up to four clusters are observed in stable states. The transition to synchronization with feedback occurs by the increase in the size of one cluster at the expense of the others. PMID- 11384391 TI - Mutual information of population codes and distance measures in probability space. AB - We studied the mutual information between a stimulus and a system consisting of stochastic, statistically independent elements that respond to a stimulus. Using statistical mechanical methods the properties of the mutual information (MI) in the limit of a large system size N are calculated. For continuous valued stimuli, the MI increases logarithmically with N and is related to the log of the Fisher information of the system. For discrete stimuli the MI saturates exponentially with N. We find that the exponent of saturation of the MI is the Chernoff distance between response probabilities that are induced by different stimuli. PMID- 11384392 TI - Time resolved collapse of a folding protein observed with small angle x-ray scattering. AB - High-intensity, "pink" beam from an undulator was used in conjunction with microfabricated rapid-fluid mixing devices to monitor the early events in protein folding with time resolved small angle x-ray scattering. This Letter describes recent work on the protein bovine beta-lactoglobulin where collapse from an expanded to a compact set of states was directly observed on the millisecond time scale. The role of chain collapse, one of the initial stages of protein folding, is not currently understood. The characterization of transient, compact states is vital in assessing the validity of theories and models of the folding process. PMID- 11384393 TI - Long-range reactive dynamics in myoglobin. AB - We report the complete vibrational spectrum of the probe nucleus 57Fe at the oxygen-binding site of the protein myoglobin. The Fe-pyrrole nitrogen stretching modes of the heme group, identified here, probe asymmetric interactions with the protein environment. Collective oscillations of the polypeptide, rather than localized heme vibrations, dominate the low frequency region. We conclude that the heme "doming" mode is significantly delocalized, so that distant sites respond to oxygen binding on vibrational time scales. This has ramifications for understanding long-range interactions in biomolecules, such as those that mediate cooperativity in allosteric proteins. PMID- 11384394 TI - Effective-area elasticity and tension of micromanipulated membranes. AB - We evaluate the effective Hamiltonian governing, at the optically resolved scale, the elastic properties of micromanipulated membranes. We identify floppy, entropic-tense and stretched-tense regimes, representing different behaviors of the effective-area elasticity of the membrane. The corresponding effective tension depends on the microscopic parameters (total area, bending rigidity) and on the optically visible area, which is controlled by the imposed external constraints. We successfully compare our predictions with recent data on micropipette experiments. PMID- 11384395 TI - Antiferroelectricity in liquid crystals. PMID- 11384397 TI - Comment on "High pressure behavior of ZrW2O8: Gruneisen parameter and thermal properties". PMID- 11384399 TI - Comment on "Invalidation of the Kelvin force in ferrofluids". PMID- 11384401 TI - Mathematical analysis of coupled parallel simulations. AB - A set of parallel replicas of a single simulation can be statistically coupled to closely approximate long trajectories. In many cases, this produces nearly linear speedup over a single simulation ( M times faster with M simulations), rendering previously intractable problems within reach of large computer clusters. Interestingly, by varying the coupling of the parallel simulations, it is possible in some systems to obtain greater than linear speedup. The methods are generalizable to any search algorithm with long residence times in intermediate states. PMID- 11384402 TI - Decoherence, pointer engineering, and quantum state protection. AB - We present a proposal for protecting states against decoherence, based on the engineering of pointer states. We apply this procedure to the vibrational motion of a trapped ion, and show how to protect qubits, squeezed states, approximate phase eigenstates, and superpositions of coherent states. PMID- 11384403 TI - Supersymmetric ratchets. AB - The overdamped Brownian motion in a periodic potential under far from equilibrium conditions is considered. A large class of systems with an intrinsic asymmetry, called supersymmetric ratchets, is identified for which the occurrence of directed transport can be ruled out without any fine-tuning of parameters. PMID- 11384404 TI - Absolute frequency measurements of the Hg+ and Ca optical clock transitions with a femtosecond laser. AB - The frequency comb created by a femtosecond mode-locked laser and a microstructured fiber is used to phase coherently measure the frequencies of both the Hg+ and Ca optical standards with respect to the SI second. We find the transition frequencies to be f(Hg) = 1 064 721 609 899 143(10) Hz and f(Ca) = 455 986 240 494 158(26) Hz, respectively. In addition to the unprecedented precision demonstrated here, this work is the precursor to all-optical atomic clocks based on the Hg+ and Ca standards. Furthermore, when combined with previous measurements, we find no time variations of these atomic frequencies within the uncertainties of the absolute value of( partial differential f(Ca)/ partial differential t)/f(Ca) < or =8 x 10(-14) yr(-1) and the absolute value of(partial differential f(Hg)/ partial differential t)/f(Hg) < or =30 x 10(-14) yr(-1). PMID- 11384406 TI - Implications of muon anomalous magnetic moment for supersymmetric dark matter. AB - The anomalous magnetic moment of the muon has recently been measured to be in conflict with the standard model prediction with an excess of 2.6sigma. Taking the excess at face value as a measurement of the supersymmetric contribution, we find that at 95% confidence level it imposes an upper bound of 500 GeV on the neutralino mass and forbids Higgsinos as being the bulk of cold dark matter. Other implications for the astrophysical detection of neutralinos include an accessible minimum direct detection rate, lower bounds on the indirect detection rate of neutrinos from the Sun and the Earth, and a suppression of the intensity of gamma ray lines from neutralino annihilations in the galactic halo. PMID- 11384407 TI - The quark condensate from K(e(4)) decays. AB - We show that, independently of the size of the quark condensate, chiral symmetry correlates the two S-wave pipi scattering lengths. In view of this constraint, the new precision data on K(e(4)) decay allow a remarkably accurate determination of these quantities. The result confirms the hypothesis that the quark condensate is the leading order parameter. PMID- 11384408 TI - Chiral symmetry and the intrinsic structure of the nucleon. AB - Understanding hadron structure within the framework of QCD is an extremely challenging problem. In order to solve it, it is vital that our thinking should be guided by the best available insight. Our purpose here is to explain the model independent consequences of the approximate chiral symmetry of QCD for two famous results concerning the structure of the nucleon. We show that both the apparent success of the constituent quark model in reproducing the ratio of the proton to neutron magnetic moments and the apparent success of the Foldy term in reproducing the observed charge radius of the neutron are coincidental. That is, a relatively small change of the current quark mass would spoil both results. PMID- 11384409 TI - Isospin asymmetry in the pseudospin dynamical symmetry. AB - Pseudospin symmetry in nuclei is investigated considering the Dirac equation with a Lorentz structured Woods-Saxon potential. The isospin correlation of the energy splittings of pseudospin partners with the nuclear potential parameters is studied. We show that, in an isotopic chain, the pseudospin symmetry is better realized for neutrons than for protons. This behavior comes from balance effects among the central nuclear potential parameters. In general, we found an isospin asymmetry of the nuclear pseudospin interaction, opposed to the nuclear spin orbit interaction which is quasi-isospin symmetric. PMID- 11384405 TI - Bounds on the CP asymmetry in like-sign dileptons from B(0)B*(0) meson decays. AB - We have measured the charge asymmetry in like-sign dilepton yields from B(0)B*(0) meson decays using the CLEO detector at the Cornell Electron Storage Ring. We find a(0)(ll) identical with[N(l(+)l(+))-N(l(-)l(-))]/[N(l(+)l(+))+N(l(-)l(-))] = +0.013+/-0.050+/-0.005. We combine this result with a previous, independent measurement and obtain Re(epsilon(B))/(1+ the absolute value of epsilon(B)(2)) = +0.0035+/-0.0103+/-0.0015 (uncertainties are statistical and systematic, respectively) for the CP impurity parameter, epsilon(B). PMID- 11384410 TI - Observation of rho/omega meson modification in nuclear matter. AB - We measured the invariant mass spectra of electron-positron pairs in the target rapidity region of 12-GeV p+A reactions. We have observed a significant difference in the mass spectra below the omega meson between p+C and p+Cu interactions. This difference indicates that the spectral shape of mesons is modified at normal nuclear-matter density. PMID- 11384411 TI - Isotopic scaling in nuclear reactions. AB - A three parameter scaling relationship between isotopic distributions for elements with Z< or =8 has been observed. This allows a simple description of the dependence of such distributions on the overall isospin of the system. This scaling law (termed isoscaling) applies for a variety of reaction mechanisms that are dominated by phase space, including evaporation, multifragmentation, and deeply inelastic scattering. The origins of this scaling behavior for the various reaction mechanisms are explained. For multifragmentation processes, the systematics is influenced by the density dependence of the asymmetry term of the equation of state. PMID- 11384412 TI - QED effects in Cu-like Pb recombination resonances near threshold. AB - In an electron-ion recombination study with Pb53+ dielectronic recombination resonances are found for as low as approximately 10(-3)-10(-4) eV relative energy. The resonances have been calculated by relativistic many-body perturbation theory and through comparison with experiment the Pb53+(4p(1/2) 4s(1/2)) energy splitting of approximately 118 eV is determined with an accuracy comparable to the position of the first few resonances, i.e., approximately 10( 3) eV. Such a precision provides a test of QED in a many-body environment at a level which can still not be reached in calculations. PMID- 11384413 TI - Pseudotime Schrodinger equation with absorbing potential for quantum scattering calculations. AB - The Schrodinger equation (Hpsi) (r) = [E+u(E)W(r)]psi(r) with an energy-dependent complex absorbing potential -u(E)W(r), associated with a scattering system, can be reduced for a special choice of u(E) to a harmonic inversion problem of a discrete pseudotime correlation function y(t) = phi(T)U(t)phi. An efficient formula for Green's function matrix elements is also derived. Since the exact propagation up to time 2t can be done with only approximately t real matrix vector products, this gives an unprecedently efficient scheme for accurate calculations of quantum spectra for possibly very large systems. PMID- 11384414 TI - Orientation of benzene in supersonic expansions, probed by IR-laser absorption and by molecular beam scattering. AB - This work represents the first experimental demonstration that planar molecules tend to travel as a "frisbee" when a gaseous mixture with lighter carriers expands into a vacuum, the orientation being due to collisions. The molecule is benzene, the prototype of aromatic chemistry. The demonstration is via two complementary experiments: interrogating benzene by IR-laser light and controlling its orientation by selective scattering on rare gas targets. The results cast new light on the microscopic mechanisms of collisional alignment and suggest a useful way to produce intense beams of aligned molecules, permitting studies of steric effects in gas-phase processes and in surface catalysis. PMID- 11384415 TI - Reaction imaging with interferometry. AB - We analyze scattering into a double-slit interferometer with target-fragment recoil detection as a monitor of quantum correlation and entanglement in few-body reaction amplitudes. We thus investigate two-slit interference with which-way information as an enhancement to modern reaction-fragment detection. We briefly consider charged-particle scattering with recoil-ion detection from the point of view of quantum information. PMID- 11384416 TI - Exact soliton solutions, shape changing collisions, and partially coherent solitons in coupled nonlinear Schrodinger equations. AB - We present the exact bright one-soliton and two-soliton solutions of the integrable three coupled nonlinear Schrodinger equations (3-CNLS) by using the Hirota method, and then obtain them for the general N-coupled nonlinear Schrodinger equations ( N-CNLS). It is pointed out that the underlying solitons undergo inelastic (shape changing) collisions due to intensity redistribution among the modes. We also analyze the various possibilities and conditions for such collisions to occur. Further, we report the significant fact that the various partially coherent solitons discussed in the literature are special cases of the higher order bright soliton solutions of the N-CNLS equations. PMID- 11384417 TI - Single-particle dynamics in collisionless magnetic reconnection. AB - The role of single-particle dynamics in driven magnetic reconnection in collisionless plasmas is investigated experimentally and analytically. The trapping of particle orbits in the magnetic cusp is observed to allow fast reconnection in the absence of a macroscopic current layer, at a rate identical to that of vacuum. The development of an electrostatic potential structure around the magnetic X line during reconnection is predicted theoretically and observed experimentally. PMID- 11384418 TI - Phase mixing and island saturation in Hamiltonian reconnection. AB - The nonlinear evolution of a Hamiltonian magnetic field line reconnection in a two-dimensional fluid plasma leads to a macroscopic equilibrium with a finite size island and fine-scale spatial structures. The latter arise from the phase mixing of the Lagrangian invariant fields. This equilibrium is the analog of the Bernstein-Greene-Kruskal equilibrium solution for electrostatic Langmuir waves. PMID- 11384419 TI - Structure formation and tearing of an MeV cylindrical electron beam in a laser produced plasma. AB - The stability of a cylindrical, solid hot electron beam propagating in a high density plasma has been studied using a two-dimensional, hybrid Darwin code. The initially solid beam evolves into a hollow, annular beam due to the Weibel instability and generates strong magnetic fields on both sides of the annular ring. The annular structure subsequently breaks up into several beamlets via a mechanism similar to a tearing instability. It is found that the magnetic fields parallel to the direction of beam propagation also grow due to the tearing process. PMID- 11384420 TI - Error field amplification and rotation damping in tokamak plasmas. AB - Toroidal rotation is normally very weakly damped in plasmas that are magnetically confined in the nominally toroidally symmetric tokamak. However, a strong damping of toroidal rotation is observed as such plasmas approach marginal stability for perturbations that produce a kinklike distortion of the plasma. It is shown that the damping of toroidal rotation by very small departures of the magnetic field from toroidal symmetry is greatly enhanced as marginal stability is approached. The response of a plasma to perturbations is studied using a set of electrical circuit elements, which provide an equation for the rotational damping that requires minimal information about the plasma. PMID- 11384421 TI - Dilation-induced phases of gases absorbed within a bundle of carbon nanotubes. AB - A study is presented of the effects of gas (especially H2) absorption within the interstitial channels of a bundle of carbon nanotubes. The ground state of the system is determined by minimizing the total energy, which includes the molecules' interaction with the tubes, the intertube interaction, and the molecules' mutual interaction (which is screened by the tubes). The consequences of swelling include a reduced threshold pressure for gas uptake and a 2.7% increase in the tubes' breathing mode frequency. PMID- 11384422 TI - Quantitative atomic-scale analysis of interface structures: transmission electron microscopy and local density functional theory. AB - Transmission electron microscopy (TEM) and local density functional theory (LDFT) are combined to analyze the microscopic structure of the rhombohedral twin interface in alpha-Al2O3. LDFT provides interfacial energetics and atomic and electronic structures for three competing models. With high-resolution TEM the atomic structure at the interface is imaged quantitatively along two orthogonal zone axes. Electron energy loss spectroscopy in TEM with nanoscale spatial resolution yields the interfacial electronic structure. Both experiments confirm the theoretically preferred model quantitatively. PMID- 11384423 TI - Controlling crystal surface termination by cleavage direction. AB - We have investigated the cleaving behavior of potassium bichromate (K(2)Cr(2)O(7)) crystals using atomic force microscopy. This crystal has a double layered AB structure along [001]. We find that, upon cleavage along the [001] plane in the <100> directions, one side is completely A terminated, while the other is B terminated. Moreover, the cleavage plane (between an A and a B layer, or between B and A) depends on the imposed direction of cleavage, i.e., [100] or [*100]. This means that the molecular layer that terminates the crystal surface can be controlled by choosing the macroscopic direction of the cleavage force. One of the two terminations is metastable and partly reconstructs to the stable termination. PMID- 11384425 TI - Minimum spanning trees on random networks. AB - We show that the geometry of minimum spanning trees (MST) on random graphs is universal. Because of this geometric universality, we are able to characterize the energy of MST using a scaling distribution [P(epsilon)] found using uniform disorder. We show that the MST energy for other disorder distributions is simply related to P(epsilon). We discuss the relationship to invasion percolation, to the directed polymer in a random media, to uniform spanning trees, and also the implications for the broader issue of universality in disordered systems. PMID- 11384424 TI - Simple "kink" model of melt intercalation in polymer-clay nanocomposites. AB - We propose a simple semiphenomenological model to describe the dynamics of polymer melt intercalation in the gallery between the adjacent clay sheets in polymer-clay nanocomposites. Within this model, the intercalation process is driven by the motion of localized excitations ("kinks") which open up the tip between the clay sheets. These kinks belong to a novel type of solitonlike excitations that appear due to the interplay between the double-well potential of the clay-clay long-range interaction, bending elasticity of the sheets, and external shear force. We find that the kink solutions can exist only if applied shear is sufficiently strong, in a qualitative agreement with experimental data. PMID- 11384426 TI - Nucleation of water and methanol droplets on cations and anions: the sign preference. AB - The barrier height to cation- and anion-induced nucleation to produce water and methanol droplets are calculated by means of an umbrella-sampling Monte Carlo method. The computer simulation corroborates the century-old finding of Wilson that the anion is a better nucleator to produce water droplets than the cation having the same magnitude of charge, even without the presence of external electric field. The simulation also shows that the cation is a better nucleator to produce methanol droplets than the anion. PMID- 11384427 TI - Solidification of a supercooled liquid in a narrow channel. AB - We simulate solidification in a narrow channel through the use of a phase-field model with an adaptive grid. In different regimes, we find that the solid can grow in fingerlike steady-state shapes, or become unstable, exhibiting unsteady growth. At low melt undercoolings, we find good agreement between our results, theoretical predictions, and experiment. For high undercoolings, we report evidence for a new stable steady-state finger shape which exists in experimentally accessible ranges for typical materials. PMID- 11384428 TI - Spatial variation of au coverage as the driving force for nanoscopic pattern formation. AB - Au induced faceting of a 4 degrees vicinal Si(001) surface was studied with chemical resolution using soft x-ray photoemission electron microscopy. For the first time a direct and quantitative determination of the local Au coverage in situ and during deposition was possible. Au atoms, necessary for the expansion of (001) terraces, are accummulated from a lattice gas, resulting in a phase separation between Au enriched terraces and Au depleted step bunches. During a second stage Au also adsorbs on the step bunches and transforms them into (119) facets. A simple Monte Carlo simulation shows that the initial coverage difference between terraces and bunches determines the regularity of the formed mesoscopic grating. PMID- 11384429 TI - Quantum diffusion of H/Ni(111) through a Monte Carlo wave function formalism. AB - We consider a quantum system coupled to a dissipative background with many degrees of freedom using the Monte Carlo wave function method. Instead of dealing with a density matrix which can be very highly dimensional, the method consists of integrating a stochastic Schrodinger equation with a non-Hermitian damping term in the evolution operator, and with random quantum jumps. The method is applied to the diffusion of hydrogen on the Ni(111) surface below 100 K. We show that the recent experimental diffusion data for this system can be understood through an interband activation process, followed by quantum tunneling. PMID- 11384430 TI - Ladder operator for the one-dimensional Hubbard model. AB - The one-dimensional Hubbard model is integrable in the sense that it has an infinite family of conserved currents. We explicitly construct a ladder operator which can be used to iteratively generate all of the conserved current operators. This construction is different from that used for Lorentz invariant systems such as the Heisenberg model. The Hubbard model is not Lorentz invariant, due to the separation of spin and charge excitations. The ladder operator is obtained by a very general formalism which is applicable to any model that can be derived from a solution of the Yang-Baxter equation. PMID- 11384431 TI - Ab initio fermi surface calculation for charge-density wave instability in transition metal oxide bronzes. AB - The electronic structure of the charge density wave (CDW) bronze (PO2)(4)(WO3)(2m), m = 4, is determined using ab initio density functional theory. The calculation shows that the Fermi surface (FS) consists in the superposition of three one-dimensional FS's associated with three types of chains. The q dependence of the electronic response function calculated from the electronic structure quantitatively accounts for the anisotropy of the fluctuations probed by x-ray diffuse scattering. The results validate the hidden nesting mechanism proposed for the CDW transitions in this series of bronzes. PMID- 11384432 TI - Aharonov-Bohm cages in 2D normal metal networks. AB - We report on magnetoresistance transport measurements performed on a bipartite tiling of rhombus in the GaAs/GaAlAs system. We observe for the first time large amplitude h/e oscillations in this network as compared to the one measured in square lattices of similar size. These oscillations are the signature of a recently predicted localization phenomenon induced by Aharonov-Bohm interferences in this peculiar network. PMID- 11384433 TI - Quantum well behavior without confining barrier observed via dynamically screened photon field. AB - Angle-resolved photoemission spectra from Na adlayers on Al(111) reveal features which behave like quantum well resonances although the substrate provides no confining barrier. These features are observed in a narrow photon energy range where overlayer collective excitations cause resonant enhancement of the photoemission intensity. The quantum well behavior is shown to be due to surface resonances of the Na/Al system. The resonances are observable using photoemission because of spatial confinement and dynamical enhancement of the local electric field within the Na films. PMID- 11384434 TI - Indirect interaction of solid-state qubits via two-dimensional electron gas. AB - We propose a mechanism of long-range coherent coupling between nuclear spin qubits in semiconductor-heterojunction quantum information processing devices. The coupling is via localized donor electrons which interact with the two dimensional electron gas. An effective interaction Hamiltonian is derived and the coupling strength is evaluated. We also discuss mechanisms of decoherence and consider gate control of the interaction between qubits. The resulting quantum computing scheme retains all the control and measurement aspects of earlier approaches, but allows qubit spacing at distances of the order of 100 nm, attainable with the present-day semiconductor device technologies. PMID- 11384435 TI - Correlation between quantized electronic states and oscillatory thickness relaxations of 2D Pb islands on Si(111)-(7 x 7) surfaces. AB - Two-dimensional lead (Pb) islands of varying heights have been grown on the Si(111)-(7 x 7) surface at low temperature. Individual islands are investigated concurrently with real-space and local-probe scanning tunneling microscopy and spectroscopy. Quantum size effects, manifested in the formation of new electronic bound states, redistribution of surface charge density, and oscillatory relaxations in island thickness are found to be perfectly correlated to each other. PMID- 11384436 TI - Coulomb blockade and coherent single-cooper-pair tunneling in single Josephson junctions. AB - We have measured the current-voltage characteristics of small-capacitance single Josephson junctions at low temperatures ( T< or =0.04 K), where the strength of the coupling between the single junction and the electromagnetic environment was controlled with one-dimensional arrays of dc SQUIDs. We have clearly observed Coulomb blockade of Cooper-pair tunneling and even a region of negative differential resistance, when the zero-bias resistance of the SQUID arrays is much higher than the quantum resistance h/e(2) approximately 26 kOmega. The negative differential resistance is evidence of coherent single-Cooper-pair tunneling in the single Josephson junction. PMID- 11384437 TI - High frequency conductivity in the quantum hall regime. AB - We have measured the complex conductivity sigma(xx) of a two-dimensional electron system in the quantum Hall regime up to frequencies of 6 GHz at electron temperatures below 100 mK. Using both its imaginary and real part we show that sigma(xx) can be scaled to a single function for different frequencies and several transitions between plateaus in the quantum Hall effect. Additionally, the conductivity in the variable-range hopping regime is used for a direct evaluation of the localization length xi. Even for large filling factor distances deltanu from the critical point we find xi approximately equals deltanu(-gamma) with a scaling exponent gamma = 2.3. PMID- 11384438 TI - Fano and Kondo resonance in electronic current through nanodevices. AB - Electronic transport through a quantum dot strongly coupled to electrodes is studied within a model with two conduction channels. It is shown that multiple scattering and interference of transmitted waves through both channels lead to Fano resonance associated with Kondo resonance. Interference effects are also pronouncedly seen in transport through the Aharonov-Bohm ring with the Kondo dot, where the current characteristics continuously evolve with the magnetic flux. PMID- 11384439 TI - Vortex collisions: crossing or recombination? AB - We investigate the collision of two vortex lines moving with viscous dynamics and driven towards each other by an applied current. Using London theory in the approach phase we observe a nontrivial vortex conformation producing antiparallel segments; their attractive interaction triggers a violent collision. The collision region is analyzed using the time-dependent Ginzburg-Landau equation. While we find that vortices will always recombine through the exchange of segments, a crossing channel appears naturally through a double collision process. PMID- 11384440 TI - Defect-unbinding and the Bose-glass transition in layered superconductors. AB - The low-field Bose-glass transition temperature in heavy-ion irradiated Bi(2)Sr(2)CaCu(2)O(8+delta) increases progressively with increasing density n(d) of irradiation-induced columnar defects, but saturates for n(d) greater or = 1.5 x 10(9) cm(-2). The maximum Bose-glass temperature corresponds to that above which diffusion of two-dimensional pancake vortices between vortex lines becomes possible, and the "linelike" character of vortices is lost. We develop a description of the Bose-glass line that quantitatively describes experiments on crystals with widely different track densities and material parameters. PMID- 11384441 TI - Effects of point defects on the phase diagram of vortex states in high- T(c) superconductors in the B parallel to c axis. AB - The phase diagram for the vortex states of high- T(c) superconductors with point defects in the B--> parallel to c axis is drawn by large-scale Monte Carlo simulations. The vortex slush (VS) phase is found between the vortex glass (VG) and vortex liquid (VL) phases. The first-order transition between this novel normal phase and the VL phase is characterized by a sharp jump of the density of dislocations. The first-order transition between the Bragg glass (BG) and VG or VS phases is also clarified. These two transitions are compared with the melting transition between the BG and VL phases. PMID- 11384442 TI - Asymmetric field profile in Bose glass phase of irradiated YBa2Cu3O(7-delta): loss of interlayer coherence around 1/3 of matching field. AB - Magneto-optical imaging in YBa(2)Cu(3)O(7-delta) with tilted columnar defects (CD's) shows an asymmetric critical-state field profile. The observed hysteretic shift of the profile ridge (trough) from the center of the sample is explained by in-plane magnetization originating from vortex alignment along CD's. The extracted ratio of the in-plane to out-of-plane magnetization component has a maximum at 1/5 of matching field ( B(Phi)) and disappears above B(Phi)/3, suggesting a reduction of interlayer coherence well below B(Phi) in the Bose glass phase. Implications are discussed in comparison with the vortex liquid recoupling observed in irradiated Bi(2)Sr(2)CaCu(2)O(8+y). PMID- 11384443 TI - Temperature dependence of the flux line lattice transition into square symmetry in superconducting LuNi2B2C. AB - We have investigated the temperature dependence of the H parallel to c flux line lattice structural phase transition from square to hexagonal symmetry, in the tetragonal superconductor LuNi2B2C ( T(c) = 16.6 K). At temperatures below 10 K the transition onset field, H2(T), is only weakly temperature dependent. Above 10 K, H2(T) rises sharply, bending away from the upper critical field. This contradicts theoretical predictions of H2(T) merging with the upper critical field and suggests that just below the H(c2)(T) curve the flux line lattice might be hexagonal. PMID- 11384444 TI - Unconventional superconductivity in CeIrIn5 and CeCoIn5: specific heat and thermal conductivity studies. AB - Low temperature specific heat and thermal conductivity measurements on the ambient pressure heavy fermion superconductors CeIrIn5 and CeCoIn5 reveal power law temperature dependences of these quantities below T(c). The low temperature specific heat in both CeIrIn5 and CeCoIn5 includes T2 terms, consistent with the presence of nodes in the superconducting energy gap. The thermal conductivity data present a T-linear term consistent with the universal limit (CeIrIn5), and a low temperature T3 variation in the clean limit (CeCoIn5), also in accord with prediction for an unconventional superconductor with lines of nodes. PMID- 11384445 TI - Direct evidence for block-by-block growth in high-temperature superconductor ultrathin films. AB - Charge neutrality and stoichiometry impose severe restrictions on the mechanisms of epitaxial growth of complex oxides. The fundamental question arises of what is the minimum growth unit when sample thickness is reduced beyond the size of the unit cell. We have investigated the growth mechanism of YBa2Cu3O7 cuprate superconductor, using a consistent approach based on the growth of noninteger numbers of YBa2Cu3O7 layers in YBa(2)Cu(3)O(7)/PrBa(2)Cu3O7 superlattices. Ex situ chemical and structural analysis evidence a 2D block-by-block mechanism in which the minimum growth units are complete unit cell blocks, growing coherently over large lateral distances. PMID- 11384446 TI - Exact universal amplitude ratios for two-dimensional Ising models and a quantum spin chain. AB - Let f(N) and xi(-1)(N) represent, respectively, the free energy per spin and the inverse spin-spin correlation length of the critical Ising model on a N x infinity lattice, with f(N)-->f(infinity) as N-->infinity. We obtain analytic expressions for a(k) and b(k) in the expansions N( f(N)-f(infinity)) = SUM (k = 1)(infinity)a(k)/N(2k-1) and xi(-1)(N) = SUM (k = 1)(infinity)b(k)/N(2k-1) for square, honeycomb, and plane-triangular lattices, and find that b(k)/a(k) = (2(2k)-1)/(2(2k-1)-1) for all of these lattices, i.e., the amplitude ratio b(k)/a(k) is universal. We also obtain similar results for a critical quantum spin chain and find that such results could be understood from a perturbated conformal field theory. PMID- 11384447 TI - Simulations on infinite-size lattices. AB - We introduce a Monte Carlo method, as a modification of existing cluster algorithms, which allows simulations directly on systems of infinite size, and for quantum models also at beta = infinity. All two-point functions can be obtained, including dynamical information. When the number of iterations is increased, correlation functions at larger distances become available. Limits q- >0 and omega-->0 can be approached directly. As examples we calculate spectra for the d = 2 Ising model and for Heisenberg quantum spin ladders with two and four legs. PMID- 11384448 TI - Magnetic spin ladder (C5H12N)2CuBr4: high-field magnetization and scaling near quantum criticality. AB - The magnetization, M(H< or =30 T,0.7< or =T< or =300 K), of (C5H12N)2CuBr4 has been used to identify this system as an S = 1/2 Heisenberg two-leg ladder in the strong-coupling limit, J( perpendicular) = 13.3 K and J( parallel) = 3.8 K, with H(c1) = 6.6 T and H(c2) = 14.6 T. An inflection point in M(H,T = 0.7 K) at half saturation, M(s)/2, is described by an effective XXZ chain. The data exhibit universal scaling behavior in the vicinity of H(c1) and H(c2), indicating that the system is near a quantum critical point. PMID- 11384449 TI - Dynamical response of quantum spin-glass models at t = 0. AB - We study the behavior of two archetypal quantum spin glasses at T = 0 by exact diagonalization techniques: the random Ising model in a transverse field and the random Heisenberg model. The behavior of the dynamical spin response is obtained in the spin-glass ordered phase. In both models it is gapless and has the general form chi(")(omega) = qdelta(omega)+chi(")(reg)(omega), with chi(")(reg)(omega) approximately omega for the Ising and chi(")(reg)(omega) approximately const for the Heisenberg, at low frequencies. The method provides new insight to the physical nature of the low-lying excitations. PMID- 11384450 TI - Electron and nuclear spin interactions in the optical spectra of single GaAs quantum dots. AB - Fine and hyperfine splittings arising from electron, hole, and nuclear spin interactions in the magneto-optical spectra of individual localized excitons are studied. We explain the magnetic field dependence of the energy splitting through competition between Zeeman, exchange, and hyperfine interactions. An unexpectedly small hyperfine contribution to the splitting close to zero applied field is described well by the interplay between fluctuations of the hyperfine field experienced by the nuclear spin and nuclear dipole/dipole interactions. PMID- 11384451 TI - Ponderomotive acceleration of photoelectrons in surface-plasmon-assisted multiphoton photoelectric emission. AB - Photoelectrons emitted from a gold target via a surface-plasmon-assisted multiphoton photoelectric process under a femtosecond laser pulse of moderate intensity are much more energetic than in an ordinary photoeffect without electron collective excitation. The phenomenon is interpreted in terms of time dependent ponderomotive acceleration of the particles by the resonant field localized at the metal surface. The amplitude of the plasmon resonance may be directly estimated by means of the electron energy spectra. PMID- 11384452 TI - Separable states are more disordered globally than locally. AB - A remarkable feature of quantum entanglement is that an entangled state of two parties, Alice ( A) and Bob ( B), may be more disordered locally than globally. That is, S(A)>S(A,B), where S(*) is the von Neumann entropy. It is known that satisfaction of this inequality implies that a state is nonseparable. In this paper we prove the stronger result that for separable states the vector of eigenvalues of the density matrix of system AB is majorized by the vector of eigenvalues of the density matrix of system A alone. This gives a strong sense in which a separable state is more disordered globally than locally and a new necessary condition for separability of bipartite states in arbitrary dimensions. PMID- 11384453 TI - A one-way quantum computer. AB - We present a scheme of quantum computation that consists entirely of one-qubit measurements on a particular class of entangled states, the cluster states. The measurements are used to imprint a quantum logic circuit on the state, thereby destroying its entanglement at the same time. Cluster states are thus one-way quantum computers and the measurements form the program. PMID- 11384454 TI - Limits to error correction in quantum chaos. AB - We study the correction of errors that have accumulated in an entangled state of spins as a result of unknown local variations in the Zeeman energy ( B) and spin spin interaction energy ( J). A nondegenerate code with error rate kappa can recover the original state with high fidelity within a time t(R) approximately Planck's over 2pikappa(1/2)/max(B,J)-independent of the number of encoded qubits. Whether the Hamiltonian is chaotic or not does not affect this time scale, but it does affect the complexity of the error-correcting code. PMID- 11384455 TI - Autocatalytic polymerization generates persistent random walk of crawling cells. AB - The autocatalytic polymerization kinetics of the cytoskeletal actin network provides the basic mechanism for a persistent random walk of a crawling cell. It is shown that network remodeling by branching processes near the cell membrane is essential for the bimodal spatial stability of the network which induces a spontaneous breaking of isotropic cell motion. Details of the phenomena are analyzed using a simple polymerization model studied by analytical and simulation methods. PMID- 11384456 TI - Direct evidence of multifractal atmospheric cascades from planetary scales down to 1 km. AB - We use 909 satellite images spanning the scale range 1-5000 km at both visible and infrared wavelengths to show that the variability at all observed scales and at all levels of intensity is very close to that predicted for a direct multiplicative scale invariant cascade starting at planetary scales. To within 1.6%/octave in scale, the observed type of (multi)scaling is very close to that theoretically predicted for universal multifractals, including multifractal phase transitions. Because of the strong vertical stratification, the scaling cannot be isotropic; these findings thus give strong support to the anisotropic "unified scaling" model of atmospheric dynamics. PMID- 11384457 TI - Spatiotemporal communication with synchronized optical chaos. AB - We propose a model system that allows communication of spatiotemporal information using an optical chaotic carrier waveform. The system is based on broad-area nonlinear optical ring cavities, which exhibit spatiotemporal chaos in a wide parameter range. Message recovery is possible through chaotic synchronization between transmitter and receiver. Numerical simulations demonstrate the feasibility of the proposed scheme, and the benefit of the parallelism of information transfer with optical wave fronts. PMID- 11384458 TI - Comment on "orbital and intrinsic angular momentum of single photons and entangled pairs of photons generated by parametric down-conversion". PMID- 11384460 TI - Optimization with extremal dynamics. AB - We explore a new general-purpose heuristic for finding high-quality solutions to hard discrete optimization problems. The method, called extremal optimization, is inspired by self-organized criticality, a concept introduced to describe emergent complexity in physical systems. Extremal optimization successively updates extremely undesirable variables of a single suboptimal solution, assigning them new, random values. Large fluctuations ensue, efficiently exploring many local optima. We use extremal optimization to elucidate the phase transition in the 3 coloring problem, and we provide independent confirmation of previously reported extrapolations for the ground-state energy of +/-J spin glasses in d = 3 and 4. PMID- 11384461 TI - Universality class of fluctuating pulled fronts. AB - It has recently been proposed that fluctuating "pulled" fronts propagating into an unstable state should not be in the standard Kardar-Parisi-Zhang (KPZ) universality class for rough interface growth. We introduce an effective field equation for this class of problems, and show on the basis of it that noisy pulled fronts in d+1 bulk dimensions should be in the universality class of the ((d+1)+1)D KPZ equation rather than of the (d+1)D KPZ equation. Our scenario ties together a number of heretofore unexplained observations in the literature, and is supported by previous numerical results. PMID- 11384462 TI - Best possible strategy for finding ground states. AB - Finding the ground state of a system with a complex energy landscape is important for many physical problems including protein folding, spin glasses, chemical clusters, and neural networks. Such problems are usually solved by heuristic search methods whose efficacy is judged by empirical performance on selected examples. We present a proof that, within the large class of algorithms that simulate a random walk on the landscape, threshold accepting is the best possible strategy. In particular, it can perform better than simulated annealing and Tsallis statistics. Our proof is the first example of a provably optimal strategy in this area. PMID- 11384463 TI - Evolution of proto-neutron stars with quarks. AB - Neutrino fluxes from proto-neutron stars with and without quarks are studied. Observable differences become apparent after 10-20 s of evolution. Sufficiently massive stars containing negatively charged, strongly interacting, particles collapse to black holes during the first minute of evolution. Since the neutrino flux vanishes when a black hole forms, this is the most obvious signal that quarks (or other types of strange matter) have appeared. The metastability time scales for stars with quarks are intermediate between those containing hyperons and kaon condensates. PMID- 11384464 TI - Absence of a singularity in loop quantum cosmology. AB - It is shown that the cosmological singularity in isotropic minisuperspaces is naturally removed by quantum geometry. Already at the kinematical level, this is indicated by the fact that the inverse scale factor is represented by a bounded operator even though the classical quantity diverges at the initial singularity. The full demonstration comes from an analysis of quantum dynamics. Because of quantum geometry, the quantum evolution occurs in discrete time steps and does not break down when the volume becomes zero. Instead, space-time can be extended to a branch preceding the classical singularity independently of the matter coupled to the model. For large volume the correct semiclassical behavior is obtained. PMID- 11384465 TI - Phase transitions for flat anti-de Sitter black holes. AB - We reexamine the thermodynamics of anti-de Sitter (adS) black holes with Ricci flat horizons using the adS soliton as the thermal background. We find that there is a phase transition which is dependent not only on the temperature but also on the black hole area, which is an independent parameter. As in the spherical adS black hole, this phase transition is related via the adS/conformal-field-theory correspondence to a confinement-deconfinement transition in the large- N gauge theory on the conformal boundary at infinity. PMID- 11384466 TI - Behavior of boundary string field theory associated with integrable massless flow. AB - We put forward an idea that the boundary entropy associated with integrable massless flow of thermodynamic Bethe ansatz (TBA) is identified with tachyon action of boundary string field theory. We show that the temperature parametrizing a massless flow in the TBA formalism can be identified with tachyon energy for the classical action at least near the ultraviolet fixed point, i.e., the open string vacuum. PMID- 11384467 TI - Verifying the Kugo-Ojima confinement criterion in Landau gauge Yang-Mills theory. AB - Expanding the Landau gauge gluon and ghost two-point functions in a power series we investigate their infrared behavior. The corresponding powers are constrained through the ghost Dyson-Schwinger equation by exploiting multiplicative renormalizability. Without recourse to any specific truncation we demonstrate that the infrared powers of the gluon and ghost propagators are uniquely related to each other. Constraints for these powers are derived, and the resulting infrared enhancement of the ghost propagator signals that the Kugo-Ojima confinement criterion is fulfilled in Landau gauge Yang-Mills theory. PMID- 11384469 TI - Anapole moment and other constraints on the strangeness conserving Hadronic weak interaction. AB - Standard analyses of low-energy NN and nuclear parity-violating observables have been based on a pi-, rho-, and omega-exchange model capable of describing all five independent s-p partial waves. Here a parallel analysis is performed for the one-body, exchange-current, and nuclear polarization contributions to the anapole moments of 133Cs and 205Tl. The resulting constraints are not consistent, though there remains some degree of uncertainty in the nuclear structure analysis of the atomic moments. PMID- 11384470 TI - Longitudinal field modes probed by single molecules. AB - We demonstrate that a strong longitudinal, nonpropagating field is generated at the focus of a radially polarized beam mode. This field is localized in space and its energy density exceeds the energy density of the transverse field by more than a factor of 2. Single molecules with fixed absorption dipole moments are used to probe the longitudinal field. Vice versa, it is demonstrated that orientations of single molecules are efficiently mapped out in three dimensions by using a radially polarized beam as the excitation source. We also show that there is no momentum or energy transport associated with the longitudinal field. PMID- 11384471 TI - Experimental observation of superparamagnetism in manganese clusters. AB - A molecular beam of manganese clusters, Mn (n) (n = 11-99), produced at 68 K is deflected toward high field by a gradient field magnet. These results indicate that Mn (n) clusters in this size range are superparamagnetic species whose intrinsic moments can be determined within the framework of the Langevin model of paramagnetism. Local minima in per-atom magnetic moments are observed for Mn (13) and Mn (19), suggestive of an icosahedral growth sequence for the smaller size range. For larger clusters, broad oscillations in the per-atom moments are observed, with a minimum near Mn (32-37) and a maximum around Mn (50-56). PMID- 11384468 TI - Precision measurements of the lambda(+)(c) and D0 lifetimes. AB - We report new precision measurements of the lifetimes of the Lambda(+)(c) and D0 from SELEX, the charm hadroproduction experiment at Fermilab. Based upon 1630 Lambda(+)(c) and 10 210 D0 decays we observe lifetimes of tau[Lambda(+)(c)] = 198.1+/-7.0+/-5.6 fs and tau[D0] = 407.9+/-6.0+/-4.3 fs. PMID- 11384472 TI - Excitation and fragmentation mechanisms in ion-fullerene collisions. AB - Electronic and vibronic excitations as well as fragmentation mechanisms in high energy ion ( H+, C+, Ar+) fullerene collisions are investigated within a fully microscopic approach, called nonadiabatic quantum molecular dynamics. The total kinetic energy loss of the projectile depends dramatically on ion mass, but, surprisingly, does not depend on the impact velocity for all ions in a certain range. This is in striking contrast to the predictions of the "stopping power" concept of solids, but explains apparently contradicting experimental observations. Signatures for nonstatistical fragmentation mechanisms are predicted. PMID- 11384473 TI - Photocount statistics of chaotic lasers. AB - We derive the photocount statistics of the radiation emitted from a chaotic laser resonator in the regime of single-mode lasing. Random spatial variations of the resonator eigenfunctions lead to strong mode-to-mode fluctuations of the laser emission. The distribution of the mean photocount over an ensemble of modes changes qualitatively at the lasing transition, and displays up to three peaks above the lasing threshold. PMID- 11384474 TI - Stability of a charged particle in a combined Penning-Ioffe trap. AB - The axial symmetry of a familiar Penning trap is broken by adding the radial magnetic field of an Ioffe trap. Despite the resulting loss of a confinement theorem, stable orbits related to adiabatic invariants are identified, expressions are given for their frequencies, and resonances that must be avoided are characterized. It seems feasible to experimentally realize the new Penning Ioffe trap to test these theoretical predictions. It also may be possible to simultaneously confine cold positrons and antiprotons in a Penning-Ioffe trap, along with any cold antihydrogen they may form. PMID- 11384475 TI - Instability and equilibration of centrifugally stable stratified Taylor-Couette flow. AB - The flow between two concentric cylinders, V(r), is studied analytically and computationally for a fluid with stable axial density stratification. A sufficient condition for linear, inviscid instability is d(V/r)(2)/dr<0 (i.e., all anticyclonically sheared flows) rather than the Rayleigh condition for centrifugal instability, d(Vr)(2)/dr<0. This implies a far wider range of instability than previously identified. The instability persists with finite viscosity and nonlinearity, leading to chaos and fully developed turbulence through a sequence of bifurcations. Laboratory tests are feasible and desirable. PMID- 11384476 TI - Ponderomotive laser acceleration and focusing in vacuum for generation of attosecond electron bunches. AB - A novel approach for the generation of ultrabright attosecond electron bunches is proposed, based on acceleration in vacuum, by a short laser pulse. The laser pulse profile is tailored such that the electrons are both focused and accelerated by the ponderomotive force of the light. Using time-averaged equations of motion, analytical criteria for optimal regime of acceleration are found. It is shown that for realistic laser parameters, a beam with up to 10(6) particles and normalized transverse and longitudinal emittances <10(-8) m can be produced. PMID- 11384477 TI - Effect of trapped ions on shielding of a charged spherical object in a plasma. AB - Collisions have traditionally been neglected in calculating the shielding around a small spherical collector in a plasma, and the plasma flow to the collector. We show analytically that, in dusty plasmas under typical discharge conditions, ion charge-exchange collisions lead to the buildup of negative-energy trapped ions which dominate the shielding cloud in the nonlinear region near a dust grain and substantially increase the ion current to the grain, even when the mean-free path is much greater than the Debye length. PMID- 11384478 TI - Excitation of ion-acoustic-like waves by subcritical currents in a plasma having equal electron and ion temperatures. AB - The effect of a magnetic-field-aligned plasma flow with a transverse velocity gradient on the excitation of current-driven ion-acoustic-like waves in a plasma having equal electron and ion temperatures (T(e) = T(i)) was investigated experimentally. In agreement with theoretical predictions, the presence of sheared plasma flow substantially reduces the critical electron drift velocity needed to produce the ion-acoustic instability. PMID- 11384479 TI - Destabilization of internal kink modes at high frequency by energetic circulating ions. AB - A theoretical model is proposed to interpret the high-frequency fishbone instability observed in tangential neutral-beam-injection discharges in a tokamak. It is shown that, when the beam ion beta exceeds a critical value, energetic circulating ions can indeed destabilize the internal kink mode through circulation resonance at a high frequency comparable to the circulation frequency of the energetic ions. The critical beta value of the energetic ions, the real frequency, and the growth rate of the mode are in general agreement with the high frequency fishbone instability observed in experiments. PMID- 11384480 TI - Relativistic electromagnetic solitons in the electron-ion plasma. AB - We present the results of the investigation about the ion motion influence on relativistic soliton structure, and show that in the case of moving multinode solitons the effect of the ion dynamics results in the limiting of its amplitude. The constraint on the maximum amplitude corresponds to either the ion motion breaking in the low-node-number case, or to the electron trajectory self intersection in the case of high-node-number solitons. The soliton breaking leads to the generation of fast ions, and provides a novel mechanism for the ion acceleration in the plasma irradiated by the high-intensity laser pulses. PMID- 11384481 TI - Fast formation of magnetic islands in a plasma in the presence of counterstreaming electrons. AB - With the help of 2D-3V (two dimensional in space and three dimensional in velocity) Vlasov simulations we show that the magnetic field generated by the electromagnetic current filamentation instability develops magnetic islands due to the onset of a fast reconnection process that occurs on the electron dynamical time scale. This process is relevant to magnetic channel coalescence in relativistic laser plasma interactions. PMID- 11384483 TI - Two disordered phases of the beta-tin structure in binary semiconductors. AB - The Ising model on a beta-tin structure has a phase diagram containing three distinct phases involving order, disorder, and a frustrated ordering. The x-ray crystallographic observation that no ordered beta-tin phases exist at room temperature in compound semiconductors may indicate either "paramagnetic" (dynamic) or frustrated (static) disorder. Density functional pseudopotential calculations determine which regimes real materials fall into and predict that in materials such as GaSb, there may be two temperature-driven transitions between beta-tin structures: from ordered, to frustrated order, to disordered. PMID- 11384482 TI - Reduction of ion thermal diffusivity associated with the transition of the radial electric field in neutral-beam-heated plasmas in the large helical device. AB - Recent large helical device experiments revealed that the transition from ion root to electron root occurred for the first time in neutral-beam-heated discharges, where no nonthermal electrons exist. The measured values of the radial electric field were found to be in qualitative agreement with those estimated by neoclassical theory. A clear reduction of ion thermal diffusivity was observed after the mode transition from ion root to electron root as predicted by neoclassical theory when the neoclassical ion loss is more dominant than the anomalous ion loss. PMID- 11384484 TI - Criticality in one dimension with inverse square-law potentials. AB - We demonstrate that the scaled order parameter for ferromagnetic Ising and three state Potts chains with inverse square interactions exhibits a universal critical jump, in analogy with the superfluid density in helium films. Renormalization group arguments are combined with numerical simulations of systems containing up to 10(6) lattice sites to accurately determine the critical properties of these models. In strong contrast with earlier work, compelling quantitative evidence for the Kosterlitz-Thouless-like character of the phase transition is provided. PMID- 11384485 TI - Imprinted networks as chiral pumps. AB - We investigate the interaction between a chirally imprinted network and a solvent of chiral molecules. We find that a liquid-crystalline polymer network is preferentially swollen by one component of a racemic solvent. This ability to separate is linked to the chiral order parameter of the network, and can be reversibly controlled via temperature or a mechanical deformation. It is maximal near the point at which the network loses its imprinted structure. One possible practical application of this effect would be a mechanical device for sorting mixed chiral molecules. PMID- 11384486 TI - Capillary condensation in a fractal porous medium. AB - Small-angle x-ray and neutron scattering are used to characterize the surface roughness and porosity of a natural rock which are described over three decades in length scales and over nine decades in scattered intensities by a surface fractal dimension D = 2.68+/-0.03. When this porous medium is exposed to a vapor of a contrast-matched water, neutron scattering reveals that surface roughness disappears at small scales, where a Porod behavior typical of smooth interfaces is observed instead. Water-sorption measurements confirm that such interface smoothing is due predominantly to the water condensing in the most strongly curved asperities rather than covering the surface with a wetting film of uniform thickness. PMID- 11384487 TI - Instability and wavelength selection during step flow growth of metal surfaces vicinal to fcc(001). AB - We study the onset and development of ledge instabilities during growth of vicinal metal surfaces using kinetic Monte Carlo simulations. We observe the formation of periodic patterns at [110] close packed step edges on surfaces vicinal to fcc(001) under realistic molecular beam epitaxy conditions. The corresponding wavelength and its temperature dependence are studied in detail. Simulations suggest that the ledge instability on fcc(1,1,m) vicinal surfaces is controlled by the strong kink Ehrlich-Schwoebel barrier, with the wavelength determined by dimer nucleation at the step edge. Our results are in agreement with recent continuum theoretical predictions, and experiments on Cu(1,1,17) vicinal surfaces. PMID- 11384488 TI - Nonclassical behavior in the capacitance of a nanojunction. AB - The capacitance of a nanojunction formed by a scanning tunneling microscope (STM) tip and a two-dimensional gold cluster was measured through the single electron tunneling spectroscopy of a double-barrier tunnel junction. By decreasing the STM tip-cluster separation, it was observed that the capacitance first increases and then decreases at short separation. This characteristic clearly deviates from the classical behavior and provides evidence for potential quantum effects on the capacitance. PMID- 11384489 TI - Adsorption of carbon monoxide on Ni(110) above atmospheric pressure investigated with surface X-ray diffraction. AB - The crystallographic structure of CO adsorbed on Ni(110) has been investigated with x-ray diffraction at ambient pressures of CO ranging from 2.3 to 10(-10) bars. At room temperature, the high pressure and vacuum structures are identical. Above room temperature ( approximately 130 degrees C), the high pressure of CO induces a restructuring of the Ni substrate that develops strained (111) microfacets. PMID- 11384490 TI - X-ray standing wave analysis of the effect of isotopic composition on the lattice constants of Si and Ge. AB - The x-ray standing wave (XSW) technique is used to measure the isotopic mass dependence of the lattice constants of Si and Ge. Backreflection allows substrates of moderate crystallinity to be used while high order reflection yields high accuracy. The XSW, generated by the substrate, serves as a reference for the lattice planes of an epilayer of different isotopic composition. Employing XSW and photoemission, the position of the surface planes is determined from which the lattice constant difference Deltaa is calculated. Scaled to DeltaM = 1 amu we find (Deltaa/a) of -0.36x10(-5) and -0.88x10(-5) for Ge and -1.8x10( 5) and -3.0x10(-5) for Si at 300 and 30 K, respectively. PMID- 11384491 TI - Spin-polarized ground state for interacting electrons in two dimensions. AB - We study numerically the ground state magnetization for clusters of interacting electrons in two dimensions in the regime where the single particle wave functions are localized by disorder. It is found that the Coulomb interaction leads to a spontaneous ground state magnetization. For a constant electronic density, the total spin increases linearly with the number of particles, suggesting a ferromagnetic ground state in the thermodynamic limit. The magnetization is suppressed when the single particle states become delocalized. PMID- 11384492 TI - Leading temperature corrections to Fermi-liquid theory in two dimensions. AB - We calculate the basic parameters of the Fermi liquid: the scattering vertex, the Landau interaction function, the effective mass, and physical susceptibilities for a model of two-dimensional (2D) fermions with a short-ranged interaction at nonzero temperature. The leading temperature dependences of the spin components of the scattering vertex, the Landau function, and the spin susceptibility are found to be linear. T-linear terms in the effective mass and in the "charge sector" quantities are found to cancel to second order in the interaction, but the cancellation is argued not to be generic. The connection with previous studies of the 2D Fermi-liquid parameters is discussed. PMID- 11384493 TI - Exponential decay properties of Wannier functions and related quantities. AB - The spatial decay properties of Wannier functions and related quantities have been investigated using analytical and numerical methods. We find that the form of the decay is a power law times an exponential, with a particular power-law exponent that is universal for each kind of quantity. In one dimension we find an exponent of -3/4 for Wannier functions, -1/2 for the density matrix and for energy matrix elements, and -1/2 or -3/2 for different constructions of nonorthonormal Wannier-like functions. PMID- 11384494 TI - Mott-hubbard metal-insulator transition in paramagnetic V2O3: an LDA+DMFT(QMC) study. AB - The electronic properties of paramagnetic V2O3 are investigated by the computational scheme LDA+DMFT(QMC). This approach merges the local density approximation (LDA) with dynamical mean-field theory (DMFT) and uses quantum Monte Carlo simulations (QMC) to solve the effective Anderson impurity model of DMFT. Starting with the crystal structure of metallic V2O3 and insulating (V0.962Cr0.038)2O3 we find a Mott-Hubbard transition at a Coulomb interaction U approximately 5 eV. The calculated spectrum is in very good agreement with experiment. Furthermore, the orbital occupation and the spin state S = 1 determined by us agree with recent polarization dependent x-ray-absorption experiments. PMID- 11384495 TI - Light-induced metal-insulator transition in a switchable mirror. AB - Rare earth hydride films can be converted reversibly from metallic mirrors to insulating windows simply by changing the surrounding hydrogen gas pressure at room temperature. At low temperatures, in situ doping is not possible in this way as hydrogen cannot diffuse. However, our finding of persistent photoconductivity under ultraviolet illumination offers an attractive possibility to tune yttrium hydride through the T = 0 metal-insulator transition. Conductivity and Hall measurements are used to determine critical exponents. The unusually large value for the product of the static and dynamical critical exponents appears to signify the important role played by electron-electron interactions. PMID- 11384496 TI - Spectroscopic evidence for the localization of Skyrmions near nu = 1 as T --> 0. AB - Optically pumped nuclear magnetic resonance measurements of 71Ga spectra were carried out in an n-doped GaAs/Al(0.1)Ga0.9As multiple quantum well sample near the integer quantum Hall ground state nu = 1. As the temperature is lowered (down to T approximately 0.3 K), a "tilted plateau" emerges in the Knight shift data, which is a novel experimental signature of quasiparticle localization. The dependence of the spectra on both T and nu suggests that the localization is a collective process. The frozen limit spectra appear to rule out a 2D lattice of conventional Skyrmions. PMID- 11384497 TI - Role of stress on charge transfer through self-assembled alkanethiol monolayers on Au. AB - We have studied charge transfer through alkanethiol molecules self-assembled on Au(111) substrates using interfacial force microscopy. Simultaneous measurement of the tip-substrate current and the normal interfacial force reveals the critical role of tip-film contact. Measurable currents are seen only for tip applied stresses above about 20 MPa, after which the current rises exponential with stress. We suggest that charge transfer results from stress-induced band-gap states near the Fermi level in these normally highly insulating molecular films. PMID- 11384498 TI - Direct transition between a singlet Mott insulator and a superconductor. AB - We argue that a normal Fermi liquid and a singlet, spin-gapped Mott insulator cannot be continuously connected, and that some intermediate phase must intrude between them. By explicitly working out a case study where the singlet insulator is stabilized by orbital degeneracy and an inverted Hund's rule coupling, mimicking a Jahn-Teller effect, we find that the intermediate phase is a superconductor. PMID- 11384499 TI - Staggered local density of states around the vortex in underdoped cuprates. AB - We have studied a single vortex with the staggered flux (SF) core based on the SU(2) slave-boson theory of high T(c) superconductors. We find that, whereas the center in the vortex core is a SF state, as one moves away from the core center a correlated staggered modulation of the hopping amplitude chi and pairing amplitude Delta becomes predominant. We predict that in this region the local density of states exhibits staggered modulation when measured on the bonds, which may be directly detected by STM experiments. PMID- 11384500 TI - Degenerate ground state in a mesoscopic YBa2Cu3O(7-x) grain boundary Josephson junction. AB - We have measured the current-phase relationship I(varphi) of symmetric 45 degrees YBa2Cu3O7-x grain boundary Josephson junctions. Substantial deviations of the Josephson current from conventional tunnel-junction behavior have been observed: (i) The critical current exhibits, as a function of temperature T, a local minimum at a temperature T*. (ii) At T approximately T*, the first harmonic of I(phi) changes sign. (iii) For T textured NiO and untextured NiO films show exchange-bias induced in plane magnetic anisotropy of nearly equal magnitude and with the Ni moment axis being nearly parallel to the exchange bias field direction. These results represent the first observation of the key step in the exchange biasing process, namely, repopulation of the antiferromagnetic domains whose magnetization axis is closest to the exchange bias field direction. PMID- 11384506 TI - Stable quantum computation of unstable classical chaos. AB - We show on the example of the Arnold cat map that classical chaotic systems can be simulated with exponential efficiency on a quantum computer. Although classical computer errors grow exponentially with time, the quantum algorithm with moderate imperfections is able to simulate accurately the unstable chaotic classical nonlinear dynamics for long times. The algorithm can be easily implemented on systems of a few qubits. PMID- 11384507 TI - Spectral linewidths of Josephson oscillators. AB - We show that the linewidth of a Josephson flux-flow oscillator has the same functional dependence on temperature, static, and dynamic resistances as the ones of Josephson single-fluxon oscillators and small Josephson junctions. This suggests a universal formula for the linewidth of Josephson oscillators. PMID- 11384508 TI - Degree distributions of growing networks. AB - The in-degree and out-degree distributions of a growing network model are determined. The in-degree is the number of incoming links to a given node (and vice versa for out-degree). The network is built by (i) creation of new nodes which each immediately attach to a preexisting node, and (ii) creation of new links between preexisting nodes. This process naturally generates correlated in degree and out-degree distributions. When the node and link creation rates are linear functions of node degree, these distributions exhibit distinct power-law forms. By tuning the parameters in these rates to reasonable values, exponents which agree with those of the web graph are obtained. PMID- 11384509 TI - Phylogenetic analysis of methanogens from the bovine rumen. AB - BACKGROUND: Interest in methanogens from ruminants has resulted from the role of methane in global warming and from the fact that cattle typically lose 6 % of ingested energy as methane. Several species of methanogens have been isolated from ruminants. However they are difficult to culture, few have been consistently found in high numbers, and it is likely that major species of rumen methanogens are yet to be identified. RESULTS: Total DNA from clarified bovine rumen fluid was amplified using primers specific for Archaeal 16S rRNA gene sequences (rDNA). Phylogenetic analysis of 41 rDNA sequences identified three clusters of methanogens. The largest cluster contained two distinct subclusters with rDNA sequences similar to Methanobrevibacter ruminantium 16S rDNA. A second cluster contained sequences related to 16S rDNA from Methanosphaera stadtmanae, an organism not previously described in the rumen. The third cluster contained rDNA sequences that may form a novel group of rumen methanogens. CONCLUSIONS: The current set of 16S rRNA hybridization probes targeting methanogenic Archaea does not cover the phylogenetic diversity present in the rumen and possibly other gastro-intestinal tract environments. New probes and quantitative PCR assays are needed to determine the distribution of the newly identified methanogen clusters in rumen microbial communities. PMID- 11384510 TI - Calcium-mediated transductive systems and functionally active gap junctions in astrocyte-like GL15 cells. AB - BACKGROUND: It has been proposed that GL15, a human cell line derived from glioblastoma multiforme, is a possible astroglial-like cell model, based on the presence of cytoplasmic glial fibrillary acidic protein. RESULTS: The aim of this work was to delineate the functional characteristics of GL15 cells using various experimental approaches, including the study of morphology, mechanism of induction of intracellular Ca2+ increase by different physiological agonists, and the presence and permeability of the gap-junction system during cell differentiation. Immunostaining experiments showed the presence and localization of specific glial markers, such as glial fibrillary acidic protein and S100B, and the lack of the neuronal marker S100A. Notably, all the Ca2+ pathways present in astrocytes were detected in GL15 cells. In particular, oscillations in intracellular Ca2+ levels were recorded either spontaneously, or in the presence of ATP or glutamate (but not KCl). Immunolabelling assays and confocal microscopy, substantiated by Western blot analyses, revealed the presence of connexin43, a subunit of astrocyte gap-junction channels. The protein is organised in characteristic spots on the plasma membrane at cell-cell contact regions, and its presence and distribution depends on the differentiative status of the cell. Finally, a microinjection/dye-transfer assay, employed to determine gap-junction functionality, clearly demonstrated that the cells were functionally coupled, albeit to varying degrees, in differentiated and undifferentiated phenotypes. CONCLUSIONS: In conclusion, results from this study support the use of the GL15 cell line as a suitable in vitro astrocyte model, which provides a valuable guide for studying glial physiological features at various differentiation phases. PMID- 11384511 TI - Seasonal variation in host susceptibility and cycles of certain infectious diseases. AB - Seasonal cycles of infectious diseases have been variously attributed to changes in atmospheric conditions, the prevalence or virulence of the pathogen, or the behavior of the host. Some observations about seasonality are difficult to reconcile with these explanations. These include the simultaneous appearance of outbreaks across widespread geographic regions of the same latitude; the detection of pathogens in the off-season without epidemic spread; and the consistency of seasonal changes, despite wide variations in weather and human behavior. In contrast, an increase in susceptibility of the host population, perhaps linked to the annual light/dark cycle and mediated by the pattern of melatonin secretion, might account for many heretofore unexplained features of infectious disease seasonality. Ample evidence indicates that photoperiod-driven physiologic changes are typical in mammalian species, including some in humans. If such physiologic changes underlie human resistance to infectious diseases for large portions of the year and the changes can be identified and modified, the therapeutic and preventive implications may be considerable. PMID- 11384512 TI - Cryptococcus neoformans infection in organ transplant recipients: variables influencing clinical characteristics and outcome. AB - Unique clinical characteristics and other variables influencing the outcome of Cryptococcus neoformans infection in organ transplant recipients have not been well defined. From a review of published reports, we found that C. neoformans infection was documented in 2.8% of organ transplant recipients (overall death rate 42%). The type of primary immunosuppressive agent used in transplantation influenced the predominant clinical manifestation of cryptococcosis. Patients receiving tacrolimus were significantly less likely to have central nervous system involvement (78% versus 11%, p =0.001) and more likely to have skin, soft tissue, and osteoarticular involvement (66% versus 21%, p = 0.006) than patients receiving nontacrolimus- based immunosuppression. Renal failure at admission was the only independently significant predictor of death in these patients (odds ratio 16.4, 95% CI 1.9-143, p = 0.004). Hypotheses based on these data may elucidate the pathogenesis and may ultimately guide the management of C. neoformans infection in organ transplant recipients. PMID- 11384513 TI - PulseNet: the molecular subtyping network for foodborne bacterial disease surveillance, United States. AB - PulseNet, the national molecular subtyping network for foodborne disease surveillance, was established by the Centers for Disease Control and Prevention and several state health department laboratories to facilitate subtyping bacterial foodborne pathogens for epidemiologic purposes. PulseNet, which began in 1996 with 10 laboratories typing a single pathogen (Escherichia coli O157:H7), now includes 46 state and 2 local public health laboratories and the food safety laboratories of the U.S. Food and Drug Administration and the U.S. Department of Agriculture. Four foodborne pathogens (E. coli O157:H7; nontyphoidal Salmonella serotypes, Listeria monocytogenes and Shigella) are being subtyped, and other bacterial, viral, and parasitic organisms will be added soon. PMID- 11384514 TI - Spoligotype database of Mycobacterium tuberculosis: biogeographic distribution of shared types and epidemiologic and phylogenetic perspectives. AB - We give an update on the worldwide spoligotype database, which now contains 3,319 spoligotype patterns of Mycobacterium tuberculosis in 47 countries, with 259 shared types, i.e., identical spoligotypes shared by two or more patient isolates. The 259 shared types contained a total of 2,779 (84%) of all the isolates. Seven major genetic groups represented 37% of all clustered isolates. Two types (119 and 137) were found almost exclusively in the USA and accounted for 9% of clustered isolates. The remaining 1,517 isolates were scattered into 252 different spoligotypes. This database constitutes a tool for pattern comparison of M. tuberculosis clinical isolates for global epidemiologic studies and phylogenetic purposes. PMID- 11384515 TI - Transmission of an arenavirus in white-throated woodrats (Neotoma albigula), southeastern Colorado, 1995-1999. AB - From 1995 to 1999, we conducted longitudinal studies of white- throated woodrats (Neotoma albigula) in southeastern Colorado. Forty-five (42.9%) of 105 female and 15 (26.8%) of 56 male N. albigula had antibodies against Whitewater Arroyo virus (WWAV). Sixteen female and three male N. albigula seroconverted during the study period, most of them during July-November, when population densities are highest. Analyses of longevity data, minimum numbers alive and infected, movements, and weight data suggest that the dominant mode of WWAV transmission among white throated woodrats in Colorado is direct contact. WWAV was recently reported to cause fatal infection in humans. Our findings will lead to better assessment of the public health threat posed by infected woodrats and may be useful in predicting periods of increased risk for human infection. PMID- 11384516 TI - Geographic distribution and genetic diversity of Whitewater Arroyo virus in the southwestern United States. AB - The purpose of this study was to extend our knowledge of the geographic distribution and genetic diversity of the arenavirus(es) associated with Neotoma species (woodrats) in the southwestern United States. Infectious arenavirus was recovered from 14 (3.3%) of 425 woodrats. The virus-positive species included N. albigula in New Mexico and Oklahoma, N. cinerea in Utah, N. mexicana in New Mexico and Utah, and N. micropus in Texas. Analyses of viral nucleocapsid protein gene sequence data indicated that all the isolates were strains of the Whitewater Arroyo virus, an arenavirus previously known only from northwestern New Mexico. Analyses of the sequence data also indicated that there can be substantial genetic diversity among strains of Whitewater Arroyo virus from conspecific woodrats collected from different localities and substantial genetic diversity among strains from different woodrat species collected from the same locality. PMID- 11384517 TI - Is high prevalence of Echinococcus multilocularis in wild and domestic animals associated with disease incidence in humans? AB - We investigated a focus of highly endemic Echinococcus multilocularis infection to assess persistence of high endemicity in rural rodents, explore potential for parasite transmission to domestic carnivores, and assess (serologically) putative exposure versus infection frequency in inhabitants of the region. From spring 1993 to spring 1998, the prevalence of E. multilocularis in rodents was 9% to 39% for Arvicola terrestris and 10% to 21% for Microtus arvalis. From June 1996 to October 1997, 6 (7%) of 86 feral dogs and 1 of 33 cats living close to the region tested positive for intestinal E. multilocularis infection. Testing included egg detection by coproscopy, antigen detection by enzyme-linked immunosorbent assay (ELISA), and specific parasite DNA amplification by polymerase chain reaction. Thus, the presence of infected domestic carnivores can increase E. multilocularis exposure risk in humans. A seroepidemiologic survey of 2,943 blood donors in the area used specific Em2-ELISA. Comparative statistical analyses of seroprevalence and clinical incidence showed an increase in Em2-seroprevalence from 1986 and 1996-97 but no increase in clinical incidence of alveolar hydatid disease. PMID- 11384518 TI - Goat-associated Q fever: a new disease in Newfoundland. AB - In the spring of 1999 in rural Newfoundland, abortions in goats were associated with illness in goat workers. An epidemiologic investigation and a serologic survey were conducted in April 1999 to determine the number of infections, nature of illness, and risk factors for infection. Thirty-seven percent of the outbreak cohort had antibody titers to phase II Coxiella burnetii antigen >1:64, suggesting recent infection. The predominant clinical manifestation of Q fever was an acute febrile illness. Independent risk factors for infection included contact with goat placenta, smoking tobacco, and eating cheese made from pasteurized goat milk. This outbreak raises questions about management of such outbreaks, interprovincial sale and movement of domestic ungulates, and the need for discussion between public health practitioners and the dairy industry on control of this highly infectious organism. PMID- 11384519 TI - Molecular epidemiology of serogroup a meningitis in Moscow, 1969 to 1997. AB - Molecular analysis of 103 serogroup A Neisseria meningitidis strains isolated in Moscow from 1969 to 1997 showed that four independent clonal groupings were responsible for successive waves of meningococcal disease. An epidemic from 1969 to the mid-1970s was caused by genocloud 2 of subgroup III, possibly imported from China. Subsequent endemic disease through the early 1990s was caused by subgroup X and then by subgroup VI, which has also caused endemic disease elsewhere in eastern Europe. A 1996 epidemic was part of the pandemic spread from Asia of genocloud 8 of subgroup III. Recent genocloud 8 epidemic disease in Moscow may represent an early warning for spread of these bacteria to other countries in Europe. PMID- 11384520 TI - Melioidosis: an emerging infection in Taiwan? AB - From January 1982 to May 2000, 17 infections caused by Burkholderia pseudomallei were diagnosed in 15 patients in Taiwan; almost all the infections were diagnosed from 1994 to May 2000. Of the 15 patients, 9 (60%) had underlying diseases, and 10 (67%) had bacteremic pneumonia. Thirteen (76%) episodes of infection were considered indigenous. Four patients died of melioidosis. Seventeen B. pseudomallei isolates, recovered from eight patients from November 1996 to May 2000, were analyzed to determine their in vitro susceptibilities to 14 antimicrobial agents, cellular fatty acid and biochemical reaction profiles, and randomly amplified polymorphic DNA patterns. Eight strains (highly related isolates) were identified. All isolates were arabinose non-assimilators and were susceptible to amoxicillin-clavulanate, piperacillin-tazobactam, imipenem, and meropenem. No spread of the strain was documented. PMID- 11384521 TI - Outbreak of human monkeypox, Democratic Republic of Congo, 1996 to 1997. AB - Human monkeypox is a zoonotic smallpox-like disease caused by an orthopoxvirus of interhuman transmissibility too low to sustain spread in susceptible populations. In February 1997, 88 cases of febrile pustular rash were identified for the previous 12 months in 12 villages of the Katako-Kombe Health Zone, Democratic Republic of Congo (attack rate = 22 per 1,000; case-fatality rate = 3.7%). Seven were active cases confirmed by virus isolation. Orthopoxvirus- neutralizing antibodies were detected in 54% of 72 patients who provided serum and 25% of 59 wild-caught animals, mainly squirrels. Hemagglutination-inhibition assays and Western blotting detected antibodies in 68% and 73% of patients, respectively. Vaccinia vaccination, which protects against monkeypox, ceased by 1983 after global smallpox eradication, leading to an increase in the proportion of susceptible people. PMID- 11384522 TI - Nipah virus infection in bats (order Chiroptera) in peninsular Malaysia. AB - Nipah virus, family Paramyxoviridae, caused disease in pigs and humans in peninsular Malaysia in 1998-99. Because Nipah virus appears closely related to Hendra virus, wildlife surveillance focused primarily on pteropid bats (suborder Megachiroptera), a natural host of Hendra virus in Australia. We collected 324 bats from 14 species on peninsular Malaysia. Neutralizing antibodies to Nipah virus were demonstrated in five species, suggesting widespread infection in bat populations in peninsular Malaysia. PMID- 11384523 TI - Third-generation cephalosporin resistance in Shigella sonnei, Argentina. AB - Shigella sonnei resistant to cefotaxime (but not to ceftazidime) was isolated for the first time in stool samples from a pediatric patient with vomiting and bloody diarrhea in northern Argentina. Microbiologic and biochemical tests confirmed the presence of an extended spectrum beta-lactamase displaying an apparent isoelectric point value of 8.2. PMID- 11384524 TI - Expanding drug resistance through integron acquisition by IncFI plasmids of Salmonella enterica Typhimurium. AB - We conducted a 30-year retrospective analysis of IncFI plasmids from Salmonella enterica serotype Typhimurium. These plasmids have been associated with the emergence of epidemic clones of multidrug-resistant Salmonella. Molecular and genetic evidence indicates that IncFI plasmids are evolving through sequential acquisition of integrons carrying different arrays of antibiotic- resistance genes. PMID- 11384525 TI - Decreased susceptibility to ciprofloxacin in Salmonella enterica serotype typhi, United Kingdom. AB - In 1999, 23% of Salmonella enterica serotype Typhi isolates from patients in the United Kingdom exhibited decreased susceptibility to ciprofloxacin (MIC 0.25-1.0 mg/L); more than half were also resistant to chloramphenicol, ampicillin, and trimethoprim. Increasing numbers of treatment failures have been noted. Most infections have been in patients with a recent history of travel to India and Pakistan. PMID- 11384526 TI - The first reported case of California encephalitis in more than 50 years. AB - A recent case of California encephalitis, a rare mosquito-borne viral disease, represents only the fourth ever reported and the first since the initial three cases in 1945. This case was diagnosed retrospectively on the basis of a rise in antibody titer between acute- and convalescent-phase serum samples. PMID- 11384527 TI - Reiter syndrome following protracted symptoms of Cyclospora infection. AB - Two large outbreaks of diarrheal illness associated with Cyclospora cayetanensis, a coccidian parasite, provided an opportunity to evaluate clinical syndromes associated with this enteric pathogen. Reiter syndrome, a triad of ocular inflammation, inflammatory oligoarthritis, and sterile urethritis, has been associated with enteric infections. We describe the first case of Reiter syndrome following protracted symptoms of Cyclospora infection. PMID- 11384528 TI - Presence of class I integrons in multidrug-resistant, low-prevalence Salmonella serotypes, Italy. AB - In 1997 to 1999, we detected class I integrons in multidrug-resistant isolates of Salmonella enterica serovars Anatum, Blockley, Brandenburg, Bredeney, Derby, Heidelberg, Livingstone, Newport, Ohio, Panama, Paratyphi B, Saintpaul, Sandiego, and Stanley. PMID- 11384529 TI - Risk for human tick-borne encephalitis, borrelioses, and double infection in the pre-Ural region of Russia. AB - We assessed the risk for human tick-borne encephalitis (TBE), ixodid tick-borne borrelioses, and double infection from 1994 to 1998 in Perm, which has among the highest rates of reported cases in Russia. We studied 3,473 unfed adult Ixodes persulcatus ticks collected from vegetation in natural foci and 62,816 ticks removed from humans. TBE virus and Borrelia may coexist in ticks. PMID- 11384530 TI - Outbreak of influenza in highly vaccinated crew of U.S. Navy ship. AB - An outbreak of influenza A (H3N2) occurred aboard a U.S. Navy ship in February 1996, despite 95% of the crew's having been appropriately vaccinated. Virus isolated from ill crew members was antigenically distinct from the vaccination strain. With an attack rate of 42%, this outbreak demonstrates the potential for rapid spread of influenza in a confined population and the impact subsequent illness may have upon the workplace. PMID- 11384531 TI - HIV-1 group O infection in Cameroon, 1986 to 1998. AB - We report a survey of HIV-1 group O infection in Cameroon during 1986 to 1998. The prevalence of HIV-1/O decreased from 0.6% to 0.4%, while HIV-1/M increased from 19.2% to 31.5% from 1994 to 1998. We concluded that HIV-1/O infection is stable in Cameroon and may be declining slightly. PMID- 11384532 TI - VEB-1-like extended-spectrum beta-lactamases in Pseudomonas aeruginosa, Kuwait. AB - Two clinical Pseudomonas aeruginosa isolates from patients in intensive care units in Kuwait were resistant to expanded-spectrum cephalosporins and showed a synergistic effect between ceftazidime and clavulanic acid. This is the first report of extended-spectrum enzymes from nosocomial isolates from the Middle East. PMID- 11384533 TI - Borrelia lonestari DNA in adult Amblyomma americanum ticks, Alabama. AB - Polymerase chain reaction analysis of 204 Amblyomma americanum and 28 A. maculatum ticks collected in August 1999 near the homes of patients with southern tick-associated rash illness and in control areas in Choctaw County, Alabama, showed Borrelia lonestari flagellin gene sequence from two adult A. americanum. The presence of B. lonestari in A. americanum ticks from Alabama suggests that this suspected pathogen may be widespread in the southeastern United States. PMID- 11384534 TI - Physicians' database searches as a tool for early detection of epidemics. AB - We analyzed retrospectively the use of Physician Desk Reference Database searches to identify epidemics of tularemia, nephropathy, Pogosta disease, and Lyme disease and compared the searches with mandatory laboratory reports to the National Infectious Diseases Register in Finland during 1995. Continuous recording of such searches may be a tool for early detection of epidemics. PMID- 11384536 TI - Candida dubliniensis candidemia in Australia. PMID- 11384535 TI - Filamentous phage associated with recent pandemic strains of Vibrio parahaemolyticus. AB - A group of pandemic strains of Vibrio parahaemolyticus has recently appeared in Asia and North America. We demonstrate that a filamentous phage is specifically associated with the pandemic V. parahaemolyticus strains. An open reading frame unique to the phage is a useful genetic marker to identify these strains. PMID- 11384537 TI - Characterization of a human granulocytic ehrlichiosis-like agent from Ixodes scapularis, Ontario, Canada. PMID- 11384538 TI - High prevalence of Sin Nombre virus in rodent populations, central Utah: a consequence of human disturbance? PMID- 11384539 TI - Hantavirus seroconversion of wild-caught Peromyscus during quarantine. PMID- 11384540 TI - Mycobacterium tuberculosis isolates of Beijing genotype in Thailand. PMID- 11384541 TI - Jungle yellow fever, Rio de Janeiro. PMID- 11384542 TI - Emergence of metronidazole-resistant Bacteroides fragilis, India. PMID- 11384543 TI - Proper nomenclature for the human granulocytic ehrlichiosis agent. PMID- 11384544 TI - Single nucleotide polymorphisms in Mycobacterium tuberculosis structural genes. PMID- 11384547 TI - The antibiotic food-chain gang. PMID- 11384548 TI - Rapid Polymerase Chain Reaction Detection of Bacteria and Their Antibiotic Resistance Genes: Improved Management, Better Treatment, and Less Resistance. PMID- 11384546 TI - Will avilamycin convert ziracine into zerocine? PMID- 11384549 TI - Clinical Trials Report. PMID- 11384550 TI - The Microbiology and Management of Acute and Chronic Rhinosinusitis. AB - Although most cases of rhinosinusitis are benign, the disruption of quality of life due to disease symptoms leads patients to seek early medical care. Ongoing debates dispute the definition, bacteriology, and medical management of chronic sinusitis, while the criteria for acute sinusitis are relatively well established. Chronic rhinosinusitis remains poorly categorized, and authors differ in opinions of symptoms, time course, and bacteriology of the infections, as well as proper medical management. Recent studies from the Mayo Clinic even question the idea that chronic sinusitis is a bacterial disease and document the presence of fungal pathogens that are responsible for the inflammatory reaction and mucosal response. In general, medical management is based on physiologic principles of re-establishing natural mucociliary function and restoring proper aeration of the paranasal sinuses after an inflammatory insult. Injudicious use and indiscriminate overprescription of antibiotics has fostered the rapid development of penicillin-resistant organisms over the past two decades. Drug resistant Streptococcus pneumonia and b-lactamase-producing Haemophilus influenzae and Moraxella catarrhalis are becoming the norm, forcing the need to consider alternatives in antibacterial management. Appropriate medical management of this common problem requires a systematic approach and consideration of adjunctive therapy. This article examines the current bacteriology of these common infections and reviews the management of acute and chronic infections. PMID- 11384551 TI - Etiology and Management of Acute and Recurrent Group A Streptococcal Tonsillitis. AB - Tonsillitis is one of the most prevalent infections in children and adolescents. The etiologic agents might be viral or bacterial. About 30% of cases are reported to be of bacterial origin, mainly due to group A Streptococcus (GAS). Although in most instances GAS tonsillitis is a self-limited disease, antibiotic treatment is recommended, mainly to prevent the suppurative and nonsuppurative poststreptococcal sequelae of acute rheumatic fever and to prevent glomerulonephritis. In this paper we review the current knowledge of the etiology of acute and recurrent GAS tonsillitis, with special emphasis on a recent hypothesis regarding the etiology of bacterial eradication failure. While penicillin V remains the drug of choice for acute tonsillitis, other antibiotics are being approved and recommended for particular indications in both Europe and the United States. PMID- 11384552 TI - The Role of Quinolones in Upper Respiratory Tract Infections. AB - Fluoroquinolones are widely used in clinical practice because of their advanced pharmacokinetic properties, potential activity against most bacterial species, excellent clinical responses, and few side effects. Quinolones have no role in the treatment of pharyngitis or simple otitis media. Until recently, the available fluoroquinolones were not indicated for the treatment of acute purulent sinusitis because of their perceived inactivity against Streptococcus pneumoniae. Although not generally considered to be drugs of first choice, older quinolones have efficacy similar to that of cephalosporins and b-lactams in randomized clinical trials. Well-conducted clinical trials have shown that the new fluoroquinolones are as effective as standard comparators in patients with suspected or proven acute bacterial sinusitis and may allow shorter treatment. Ciprofloxacin remains the fluoroquinolone of choice for chronic otitis media and malignant otitis media. The new "respiratory" fluoroquinolones have microbiologic and pharmacokinetic advantages over the older agents. Clinical trials have confirmed clinical activity, but superiority compared with older agents has not been conclusively shown. Trials devised to demonstrate clinical or pharmacoeconomic benefits are still required. PMID- 11384553 TI - Mycobacterial Infections of the Head and Neck. AB - Mycobacteria are important causes of head and neck infections. Mycobacterial lymphadenitis may be caused by both Mycobacterium tuberculosis and a variety of nontuberculous myocbacteria. Changes in the epidemiology of tuberculosis have caused a shift of the peak age range of tuberculous lymphadenitis from childhood to ages 20 to 40 years. Short-course chemotherapy is highly effective. Mycobacterium avium has become the most common cause of nontuberculous lymphadenitis, but new mycobacterial species are increasingly recognized. Treatment consists primarily of complete surgical excision, although roles for antimycobacterial chemotherapy are being identified. Transient flares of mycobacterial lymphadenitis, which occur during initiation of antituberculous therapy and in HIV-infected patients after initiation of antiretroviral therapy, may respond to short courses of corticosteroids. Tuberculous otitis media has become uncommon. Otitis media due to nontuberculous mycobacterial infection is increasingly seen in patients with pre-existing ear disease and after surgical and otic interventions. Tuberculosis of the eye has also become uncommon but may occur via hematogenous dissemination or direct innoculation. Nontuberculous mycobacteria, most commonly Mycobacterium chelonae and Mycobacterium fortuitum, may cause keratitis, usually after some form of corneal trauma. PMID- 11384554 TI - Respiratory Syncytial Virus: Update on Infection, Treatment, and Prevention. AB - Respiratory syncytial virus (RSV) infection, which primarily manifests as bronchiolitis or pneumonia, is the leading cause of lower respiratory tract infection in infants and young children. It is associated with more than 100,000 pediatric hospitalizations each year in the United States. Infants who were premature; have chronic lung disease, congenital heart disease, or immunodeficiency disorders; or have underlying metabolic or neuromuscular disorders are at increased risk for especially severe RSV disease. Treatment of children hospitalized with RSV disease is primarily supportive, with administration of supplemental oxygen and fluid replacement therapy. Bronchodilators may benefit at least a subset of such patients. Antiviral therapy with aerosolized ribavirin is available for high-risk, severely ill patients. Handwashing, cleaning of environmental surfaces, and cohorting in hospital settings may decrease RSV transmission. In children born premature and younger than 1 year of age, and in patients with bronchopulmonary dysplasia younger than 2 years of age, passive protection against severe RSV disease may be achieved through monthly injections of anti-RSV antibody (palivizumab) during winter months. No vaccine is available to provide active immunity against RSV, but live attenuated and subunit cloned surface protein vaccines are in development. PMID- 11384555 TI - Pulmonary Manifestations of Bioterrorism. AB - Along with smallpox, inhalation anthrax and pneumonic plague are among the diseases most likely to be spread by biowarfare, either from a rogue nation or terrorist group. Neither anthrax nor plague has been seen by many pulmonary (or any other) physicians in the United States. This article summarizes these two diseases as pulmonary manifestations of bioterrorism and discusses the possibility of avian influenza as a potential respiratory pathogen in biowarfare. It is hoped that phyisicians will need to know this information only as an academic exercise and not because of a clinical circumstance. PMID- 11384556 TI - Hantavirus Pulmonary Syndrome in the United States. AB - Since the first outbreak of hantavirus pulmonary syndrome (HPS) in 1993, understanding of the vast distribution and potential impact of hantaviruses has grown. At least 277 cases of HPS have been documented in the United States. The full clinical spectrum has yet to be elucidated, and one outbreak suggested the possibility of person-to-person transmission. New research has identified the b-3 integrins as cellular receptors for hantaviruses and has determined the pivotal role of the immune system in pathogenesis. Rapid diagnosis has been facilitated by a new immunoblot assay to detect Sin Nombre virus infection. Treatment remains primarily supportive; however, a placebo- controlled trial of ribavirin is ongoing. Extracorporeal membrane oxygenation may be a potential therapy in severe cases; inhaled nitric oxide needs further study. Vaccines developed against hantaviruses associated with hemorrhagic fever and renal syndrome might be effective against HPS-associated strains. PMID- 11384557 TI - Nosocomial Pneumonia: More Than Just Ventilator-Associated. AB - We reviewed literature published from 1995 through 2000 on developments in ventilator-associated pneumonia. There is no gold standard with which to compare the accuracy of various invasive procedures performed for diagnosis. Moreover, leaders in the field are calling for an outcomes-based analysis to assess the utility of invasive procedures. Two things are clear: 1) adequate empiric therapy is beneficial, and 2) changes in therapy based on recovery of pathogens by invasive means do not affect outcome. Clinicians are urged to review local antimicrobial resistance patterns and to initiate empiric therapy on the basis of those data. PMID- 11384558 TI - Coccidioidomycosis Pulmonary Infection. AB - Disease caused by Coccidioides immitis has increased in frequency in recent years because of marked population shifts into highly endemic areas, as well as perturbations of the environment caused by construction, agricultural, and natural activities. Because disease may occur in persons in locations outside of the endemic regions, practitioners must be aware of the possibility of infection in those who may have traveled or resided in areas of risk. Recently, advances in laboratory methods have facilitated diagnosis of coccidioidomycosis, and information about therapies that are easier to administer has become available. Challenges in the management of this infection still remain, particularly with regard to more reliable antifungal drugs and protective vaccines. PMID- 11384559 TI - Pulmonary Histoplasmosis. AB - Most patients with pulmonary histoplasmosis experience a self-limited illness that rarely requires treatment. However, patients who inhale a large burden of organisms from the environment and those who are immunosuppressed may develop severe, life-threatening pneumonia. Chronic histoplasmosis occurs almost exclusively in patients with severe chronic obstructive pulmonary disease. Complications of pulmonary histoplasmosis are mostly related to persistent mediastinal lymphadenopathy. Mild to moderate forms of pulmonary histoplasmosis should be treated with itraconazole. Patients with severe pulmonary involvement should initially receive amphotericin B; after the patient's condition has improved, therapy can be changed to itraconazole. Preventive measures should be used to protect workers from exposure to large numbers of Histoplasma capsulatum organisms. PMID- 11384560 TI - Pulmonary Nocardiosis. AB - Pulmonary nocardiosis is an uncommon but serious infection that is increasingly found in immunosuppressed persons, especially transplant recipients and persons with AIDS. The Nocardia species are denizens of soil and decaying plants that gain entry to humans through inhalation or inoculation. Pulmonary nocardiosis typically presents as an acute to subacute necrotizing pneumonia, with a variable clinical picture. Metastatic infections of the brain and subcutaneous tissues are common complications. Most clinical laboratories can isolate these microorganisms, but final speciation may be a challenge and antimicrobial susceptibility testing is especially difficult because of the slow rate of growth of Nocardia species. Full identification of species and susceptibility testing is important because of the epidemiologic implications and the difficulties of successfully treating these infections in immunosuppressed patients. Sulfonamides, including trimethoprim-sulfamethoxazole, remain the most reliable antimicrobials. Many alternative agents are active against Nocardia in vitro, but clinical data are limited. PMID- 11384561 TI - Viral and Cellular Dynamics in HIV Disease. AB - The control mechanisms that maintain a steady-state viral load during chronic HIV 1 infection are critical to understanding the pathophysiology of HIV disease. This paper compares the conceptual features of the two alternative models of viral control, target cell limitation and immune control, with available data on the viral and cellular dynamics of HIV-1 infection and the pattern of changes induced by effective antiretroviral drug therapy. These data suggest that an antigen-driven immune response is the primary control mechanism for in vivo viral growth. PMID- 11384562 TI - Immune Reconstitution Strategies in HIV. AB - Although potent antiretroviral therapy can dramatically decrease HIV replication and improve some aspects of host immunity, incomplete immune reconstitution persists even after several years of fully suppressive therapy. In addition, long term toxicities of antiretroviral medications and the probability of developing multidrug-resistant virus with long-term use indicate that alternate means of controlling viral replication are needed for more durable suppression of HIV. Immune-based therapies may help potentiate the host's own defenses against HIV and other pathogens, and may ultimately result in more durable viral suppression and lower incidence of antiretroviral therapy-related side effects and toxicities. PMID- 11384563 TI - Clinical evaluation of twice-daily emedastine 0.05% eye drops (Emadine eye drops) versus levocabastine 0.05% eye drops in patients with allergic conjunctivitis. AB - PURPOSE: The efficacy and safety of emedastine 0.05% eye drops (Emadine; Alcon Laboratories, Inc, Fort Worth, Texas), a new H(1) antagonist, were studied in comparison to levocabastine 0.05% eye drops (Livostin; Janssen-Cilag N V, Berchem, Belgium) during a twice-daily treatment schedule for 6 weeks in adult and pediatric patients with seasonal allergic conjunctivitis. METHODS: In a prospective, multicenter, randomized, double-masked, parallel group study, 222 patients with allergic conjunctivitis were randomized (221 received treatment) to either emedastine or levocabastine, instilled twice daily for 6 weeks. Patient diaries were completed four times daily (before the morning and evening instillations, at noon, and in the afternoon), and clinical examinations were conducted at regular intervals. Primary efficacy variables of ocular redness and itching and secondary efficacy variables of chemosis, eyelid swelling, patient diary data, and physician's global assessment were analyzed. RESULTS: Both emedastine and levocabastine produced a statistically significant (P =.0001) reduction in itching and redness within 5 minutes of the first instillation. All signs and symptoms improved progressively over the 6-week treatment period. After 7 days of use, and throughout the remainder of the study, emedastine was statistically superior to levocabastine (P <.006) in preventing and alleviating the signs and symptoms (itching, redness, chemosis, and eyelid swelling) of allergic conjunctivitis. CONCLUSIONS: Emedastine 0.05% eye drops administered twice daily are more efficacious than levocabastine 0.05% eye drops in the prevention and treatment of the signs and symptoms of allergic conjunctivitis in adults and children of 4 years and above. Both emedastine 0.05% eye drops and levocabastine 0.05% eye drops were well tolerated. PMID- 11384564 TI - Risk factors for progression of visual field abnormalities in normal-tension glaucoma. AB - PURPOSE: To uncover risk factors for the highly variable individual rates of progression in cases of untreated normal-tension glaucoma. METHODS: Visual field data were assembled from 160 subjects (160 eyes) enrolled in the collaborative normal-tension glaucoma study during intervals in which the eye under study was not receiving intraocular pressure-lowering treatment during prerandomization and postrandomization intervals. Analyses included multivariate analysis of time dependent Cox proportional hazard, Kaplan-Meier analysis of "survival" without an increment of visual field worsening, and comparison of slopes of change in mean deviation global index over time. RESULTS: Most migraine occurred in women, but analysis demonstrated that gender and presence of migraine contribute separately to the overall risk. The risk ratio for migraine, adjusted for the other variables was 2.58 (P =.0058), for disk hemorrhage was 2.72 (P =.0036), and for female gender 1.85 (P =.0622). The average fall in the mean deviation index was faster in nonmigrainous women than in nonmigrainous men (P =.05). Suggesting genetic influence, Asians had a slower rate of progression (P =.005), and the few black patients enrolled had a tendency for faster progression. However, self declared history of family with glaucoma or treated for glaucoma did not affect the rate of progression. Neither age nor the untreated level of intraocular pressure affected the rate of untreated disease progression, despite their known influence on prevalence. CONCLUSIONS: Whereas risk factors for prevalence help select populations within which to screen for glaucoma, the factors that affect the rate of progression help decide the expected prognosis of the individual's untreated disease and thereby the frequency of follow-up and aggressiveness of the therapy to be undertaken. PMID- 11384565 TI - Incidence and prevalence of short wavelength automated perimetry deficits in ocular hypertensive patients. AB - PURPOSE: To determine the prevalence and incidence of short wavelength automated perimetry deficits in comparison to standard automated perimetry deficits in patients with ocular hypertension. METHODS: Five hundred eyes of 250 patients with ocular hypertension were recruited into a prospective, longitudinal study and tested with standard automated perimetry and short wavelength automated perimetry annually for 5 years. Both eyes of 60 normal subjects, 21 to 85 years of age, were used to establish normative data for short wavelength automated perimetry and standard automated perimetry. This allowed independent evaluation of left and right eyes of patients. All normal data were corrected for age, and short wavelength automated perimetry results were corrected for lens transmission. The lowest fifth and first percentiles for the normal observers were derived for the 10 glaucoma hemifield test zones for short wavelength automated perimetry and standard automated perimetry. Visual fields were considered outside normal limits if two glaucoma hemifield test zones were below the normal fifth percentile or one glaucoma hemifield test zone was below the normal first percentile. RESULTS: Baseline prevalence of short wavelength automated perimetry and standard automated perimetry deficits were 9.4% and 1.4%, respectively. During the study, incident rates of field loss were 6.2% (1.23% per year) for short wavelength automated perimetry and 5.9% (1.18% per year) for standard automated perimetry. Once abnormal, 80% of short wavelength automated perimetry fields remained abnormal on the next examination, whereas only 45% of abnormal standard automated perimetry fields remained abnormal. New short wavelength automated perimetry deficits in ocular hypertensives were more prominent and more persistent than new standard automated perimetry deficits. CONCLUSIONS: Our findings are consistent with the interpretation that short wavelength automated perimetry deficits occur before standard automated perimetry deficits in glaucoma. The similar incidence rates suggest that both standard automated perimetry and short wavelength automated perimetry are monitoring the same underlying glaucomatous disease process. PMID- 11384566 TI - The effects of astigmatism and working distance on optic nerve head images using a Heidelberg Retina Tomograph scanning laser ophthalmoscope. AB - PURPOSE: To determine effects of astigmatism and working distance on optic nerve head images in normal patients using the Heidelberg Retina Tomograph. METHODS: The optic disks of 51 normal healthy subjects, aged 19 to 44 years, were imaged through dilated pupils. Subjects with 0.75 DC or less of astigmatism were imaged without correction at a working distance of 15 mm. They were then re-imaged with a cylindrical correction of +3.00 DC at 90 degrees axis (n = 20). Naturally astigmatic subjects with more than 1.00 DC were imaged without correction and then re-imaged once this was neutralized with their appropriate spectacle prescription (n = 15). The effects of working distance were studied using subjects with 0.75 DC or less (n = 16). Two working distances were used, 15 and 25 mm. At each session the means of three topographic images were taken from which standard deviations and parameters were recorded. Parameters analyzed included cup shape measure, rim area, and inferior temporal rim volume. Z-profile full width at half maximum was calculated from one image per subject for each condition. RESULTS: No significant difference was found in the measured parameters of the optic disk for any astigmatic condition or changes in working distance (P >.05), (paired t test). Both the standard deviation of the mean topographic images and the Z-profile half-maximum width of the axial intensity profile were significantly greater with induced astigmatism of +3.00 DC (P values 0.3 and.00, respectively). CONCLUSIONS: Optic disk parameters are not significantly affected by uncorrected astigmatism (up to 2.50 DC) or working distance. The algorithm used by the Heidelberg Retina Tomograph to generate topographic maps is sufficiently robust that astigmatism up to 2.50 DC does not require correction. PMID- 11384567 TI - Potential mechanism for the additivity of pilocarpine and latanoprost. AB - PURPOSE: To determine the ocular hypotensive mechanism underlying the additivity of latanoprost and pilocarpine. METHODS: This randomized, double-masked study included 30 patients with ocular hypertension on no ocular medications for at least 3 weeks. On each of six visits to the clinic, measurements were taken of aqueous flow and outflow facility by fluorophotometry, intraocular pressure by tonometry, and episcleral venous pressure by venomanometry. Uveoscleral outflow was calculated. Clinic visits were scheduled on baseline day; on day 8 of four times daily pilocarpine (2%) to one eye and vehicle to the other; on day 8 of continued pilocarpine/vehicle treatment plus latanoprost (0.005%) once daily to both eyes; after a 3-week washout period; on day 8 of once-daily latanoprost to one eye and vehicle to the other; and on day 8 of continued latanoprost/vehicle treatment plus pilocarpine four times a day to both eyes. Drug-treated eyes were compared with contralateral vehicle-treated eyes and with baseline day by paired t tests. Combined pilocarpine and latanoprost-treated eyes were compared with individual drug-treated eyes and with baseline day using the Bonferroni test. RESULTS: Compared with baseline, pilocarpine reduced intraocular pressure from 18.9 to 16.2 mm Hg (P =.001) and increased outflow facility from 0.18 to 0.23 microl per minute per mm Hg (P =.03). No other parameters were affected. Adding latanoprost further reduced intraocular pressure to 13.7 mm Hg (P <.001) and increased uveoscleral outflow from 0.82 to 1.36 microl per minute (P =.02). Latanoprost alone reduced intraocular pressure from 17.6 to 14.3 mm Hg (P <.0001) and increased uveoscleral outflow from 0.89 to 1.25 microl per minute (P =.05). Adding pilocarpine to the latanoprost treatment further reduced intraocular pressure to 12.7 mm Hg (P <.001) and increased outflow facility from 0.21 to 0.30 microl per minute per mm Hg (P =.03). CONCLUSIONS: Latanoprost and pilocarpine predominantly increase uveoscleral outflow and outflow facility, respectively, when given alone. These drugs are additive because pilocarpine does not inhibit the uveoscleral outflow increase induced by latanoprost. PMID- 11384569 TI - Role of frequency doubling perimetry in detecting neuro-ophthalmic visual field defects. AB - PURPOSE: To report the ability of frequency doubling perimetry to detect "neuro ophthalmic" field defects, characterize them as hemianopic or quadrantanopic, and differentiate glaucomatous from "other" neuro-ophthalmic field defects. METHODS: Sixty eyes of 30 normal subjects, 50 eyes of 29 patients with glaucomatous defects, and 138 eyes of 103 patients with "typical" neuro-ophthalmic field defects underwent automated perimetry using the Swedish Interactive Threshold Algorithm and frequency doubling perimetry. The sensitivity and specificity for identification of a field defect (frequency doubling perimetry 20-5 and 20-1 screening tests), or to characterize hemianopia/quadrantanopia (full threshold test) were determined. Ability to discriminate glaucomatous defects was determined by comparing frequency doubling perimetry full threshold test in glaucoma to pooled results of normal and neuro-ophthalmic groups. RESULTS: On frequency doubling perimetry, a single point depressed to less than 1% probability had a sensitivity of 97.1% (20-5 test) and 95.7% (20-1 test) for detecting a neuro-ophthalmic visual field defect. The corresponding specificities were 95% using pooled results in normal subjects and patients with glaucoma and "other" neuro-ophthalmic field defects. In 20-5 screening a single abnormal point depressed to less than 2% probability level had a sensitivity of 98.6% (specificity 85%). Two abnormal points in the 20-1 screening depressed to less than 1% probability level had a specificity of 100% (sensitivity 84.8%). In frequency doubling perimetry full threshold, sensitivity and specificity for detection of hemianopia were 86.8% and 83.2%; for quadrantanopia they were 79.2% and 38.6%. The sensitivity and specificity for categorizing a defect as glaucomatous were 86% and 74.7%. CONCLUSIONS: Frequency doubling perimetry is a sensitive and specific test for detecting "neuro-ophthalmic" field defects. The presence of two abnormal points (20-1 screening program) "rules in" the presence of a field defect. A normal 20-5 program (absence of a single abnormal point) almost "rules out" a defect. Frequency doubling perimetry could not accurately categorize hemianopic, quadrantanopic, or glaucomatous defects. PMID- 11384568 TI - Brimonidine 0.2% given two or three times daily versus timolol maleate 0.5% in primary open-angle glaucoma. AB - PURPOSE: To evaluate the efficacy and safety of brimonidine 0.2% two or three times daily versus timolol maleate 0.5% solution twice daily. METHODS: Patients with primary open-angle glaucoma were randomized by Latin square technique to one of the three treatment sequences in this crossover, prospective double-masked trial. Each treatment period consisted of 6 weeks of chronic dosing followed by a diurnal curve for the intraocular pressure measured at 08:00, 10:00, 16:00, 18:00, 20:00, 22:00, and 24:00 hours. Intraocular pressure was measured by applanation tonometry. RESULTS: Thirty patients completed this trial. The average diurnal intraocular pressures in the trial were measured for timolol maleate (17.7 +/- 2.7 mm Hg), brimonidine given three times daily (18.0 +/- 2.2 mm Hg), and brimonidine given twice daily (19.2 +/- 2.4 mm Hg). There was a statistical difference between groups (P <.005). When groups were compared by pairs, three times daily dosing with brimonidine and timolol maleate both reduced the pressure more than twice daily brimonidine at every time point past 10:00 hours and for the diurnal curve (P <.05). In contrast, three times daily brimonidine and timolol maleate were statistically similar for the diurnal pressure, and each time point, except timolol maleate, decreased the pressure more at 16:00 (P =.042). Safety was similar between groups. CONCLUSIONS: This study demonstrated that both timolol maleate twice daily and brimonidine three times daily provide a similar intraocular pressure reduction to each other. Timolol maleate twice daily and brimonidine three times daily provide a greater decrease in pressure in the late afternoon and nighttime hours, compared with brimonidine twice daily. PMID- 11384570 TI - Duration of positive urine for cocaine metabolite after ophthalmic administration: implications for testing patients with suspected Horner syndrome using ophthalmic cocaine. AB - PURPOSE: To determine the duration of positive urine for benzoylecgonine, the major metabolite excreted in the urine, after topical ophthalmic administration of cocaine as one would perform for testing the presence of Horner syndrome. METHODS: Two drops of cocaine 10% were applied to each eye of 50 normal subjects. Urine samples were collected 4 to 6, 24, 48, 72, and 96 hours later. Each sample was assayed for benzoylecgonine using a screening competitive enzyme donor immunoassay followed by a highly specific and sensitive confirmatory gas chromatography-mass spectrometry assay. We employed assays and cutoff levels that fulfilled guidelines required by the Substance Abuse and Mental Health Services of the United States Department of Health and Human Services to mimic measures that exist for federal workplace drug testing. RESULTS: Of the 50 subjects, there were 25 women and 25 men, ranging in age from 19 to 59 years (median age, 40 years). Positive screening samples were obtained in 47 of 50 subjects (94%) 4 to 6 hours, 35 of 50 (70%) 24 hours, and 1 of 50 (2%) 48 hours after topical application of cocaine. None of the samples tested positive at 72 hours or beyond. Using the confirmatory assay's cutoff as the gold standard for a positive test, the sensitivity of the screening assay was 96% 4 to 6 hours, 90% 24 hours, and 14% 48 hours after topical application. Urine benzoylecgonine concentration was negatively correlated with body mass index and positively correlated with urine creatinine concentration. CONCLUSIONS: Patients should be informed that their urine may test positive for cocaine, if assayed according to US federal guidelines and using the protocol employed in this study, up to 2 days after undergoing testing for Horner syndrome. PMID- 11384571 TI - Acrylic intraocular lens placement in conjunction with pars plana vitrectomy. AB - PURPOSE: To report the results of placing a foldable acrylic intraocular lens through a clear cornea incision in eyes undergoing pars plana vitrectomy. METHODS: This interventional case series is a retrospective report of 15 eyes of 15 consecutive patients undergoing foldable (acrylic) intraocular lens insertion in conjunction with pars plana vitrectomy in a single academic institution. Outcome measures included visual acuity and complications. RESULTS: The intraocular lens was successfully implanted and retained in position in all 15 cases. The intraocular lens did not pose difficulty in examining the fundus or cause any vitreoretinal complications. With mean follow-up of 4.5 months (range, 1--10 months), final median postoperative visual acuity range was 20/200 (range, 20/40--2/200) and consistent with the level of vitreoretinal disease. CONCLUSION: Acrylic intraocular lenses may be safely implanted in conjunction with pars plana vitrectomy in selected cases. PMID- 11384572 TI - Effect of sildenafil citrate (Viagra) on the ocular circulation. AB - PURPOSE: Sildenafil citrate induces vasodilation by enhancing the smooth muscle relaxant effects of nitric oxide. We have previously reported that nitrate compounds, a different group of nitric oxide-mediated vasodilators used mainly for the treatment of ischemic cardiac diseases, produce an increase in optic nerve head circulation and retinal venous vasodilation. The purpose of the present investigation was to evaluate the effect of sildenafil on ocular circulation. METHODS: In a double-blind, randomized, crossover trial, 15 healthy male volunteers received 100-mg doses of sildenafil citrate (Viagra; Pfizer, Inc, New York, New York) or matching placebo on 2 separate days. Laser Doppler flowmetry was used to assess foveolar choroidal and optic nerve rim circulatory parameters. Measurements were obtained in one eye at baseline, 1 hour, and 5 hours after dosing. Blood pressure and intraocular pressure were monitored, and perfusion pressure was calculated. RESULTS: Mean optic nerve head blood flow measurements at baseline, 1 hour, and 5 hours were 11.6 +/- 2.2 arbitrary units (+/- SD), 12.5 +/- 2.8, and 12.1 +/- 2.4 after sildenafil and 11.9 +/- 2.5, 12.6 +/- 3.1, and 13.0 +/- 3.0 after placebo, respectively. When compared with placebo, no significant change in mean blood pressure, intraocular pressure, perfusion pressure, or choroidal or optic nerve circulatory parameters were observed after sildenafil treatment. The power to detect a 20% change in optic nerve head and choroidal blood flow after sildenafil was approximately 90%. CONCLUSIONS: In comparison with placebo, no significant change in optic nerve rim or foveolar choroidal blood flow was observed after treatment with sildenafil. This suggests that nitrate compounds and sildenafil may differentially affect ocular circulation. Furthermore, no significant effects on intraocular pressure, systemic blood pressure, or ocular perfusion pressure were detected after sildenafil treatment. PMID- 11384573 TI - Evaluation of the retinal nerve fiber layer thickness in eyes with idiopathic macular holes. AB - PURPOSE: To compare the retinal nerve fiber layer thickness in eyes with idiopathic macular holes and age-matched normal controls using scanning laser polarimeter. METHODS: The retinal nerve fiber layer thickness was measured in 40 eyes of 40 consecutive patients with idiopathic macular hole (stage 1, 10 eyes; stage 2, eight eyes; stage 3, 15 eyes; stage 4, seven eyes) and 40 eyes of 40 age matched normal controls with a scanning laser polarimeter. The retinal nerve fiber layer thickness within a 10-pixel-wide ellipse located concentrically with the disk and located 1.5-disk diameters from the center of the disk was measured. The mean overall retinal nerve fiber layer thickness of the peripapillary retina, four 90-degree quadrants, and 16 equal sectors of every 22.5 degrees was calculated for both groups. The retinal nerve fiber layer thickness in the two groups was statistically compared. RESULTS: The mean retinal nerve fiber layer thickness measurement for the overall peripapillary retina and for three of the four 90-degree quadrants was not significantly different between the two groups. However, the temporal 90-degree quadrant was significantly thinner in the macular hole group (47.2 versus 54.6 microm, P =.026). For the 16 sectors of 22.5 degrees, the lower three sectors of the four sectors in the temporal quadrant were thinner in the macular hole group (P <.05). CONCLUSIONS: The retinal nerve fiber layer thickness of the papillomacular area is thinner in eyes with idiopathic macular hole than that in normal eyes. The progressive thinning of the retinal nerve fiber layer thickness as the stage of the macular hole advances may suggest that surgery should be done at the earliest stage. PMID- 11384574 TI - Analysis of the ABCR (ABCA4) gene in 4-aminoquinoline retinopathy: is retinal toxicity by chloroquine and hydroxychloroquine related to Stargardt disease? AB - PURPOSE: To determine if mutations in ABCR (ABCA4) are associated with chloroquine/hydroxychloroquine retinopathy. METHODS: DNA from eight patients with chloroquine or hydroxychloroquine retinopathy was studied. Controls were 80 individuals over age 65 years with normal retinal examinations. Ophthalmoscopy, color vision testing, visual fields, retinal photography, and fluorescein angiography were performed on the eight patients. Direct DNA sequencing of the exons and flanking intronic regions of the ABCR gene was completed for all patients. RESULTS: Clinical evaluation confirmed the diagnosis of chloroquine/hydroxychloroquine retinopathy and excluded Stargardt disease in each patient. Two patients had heterozygous ABCR missense mutations previously associated with Stargardt disease. None of the controls had these missense mutations. Three other patients had other missense polymorphisms. CONCLUSIONS: Some individuals who have ABCR mutations may be predisposed to develop retinal toxicity when exposed to chloroquine/hydroxychloroquine. We urge further study of a larger cohort of patients with chloroquine/hydroxychloroquine retinopathy. PMID- 11384575 TI - Local cellular sources of apolipoprotein E in the human retina and retinal pigmented epithelium: implications for the process of drusen formation. AB - PURPOSE: The inheritance of specific apolipoprotein E allelles has been linked to atherosclerosis, Alzheimer disease, and, most recently, to the incidence of age related macular degeneration. Apolipoprotein E is a common component of the extracellular plaques and deposits characteristic of these disorders, including drusen, which are a hallmark of age-related macular degeneration. Accordingly, we assessed the potential biosynthetic contribution of local ocular cell types to the apolipoprotein E found in drusen. METHODS: We measured apolipoprotein E mRNA levels in human donor tissues using a quantitative assay of apolipoprotein E transcription, and we localized apolipoprotein E protein to specific cell types and compartments in the neural retina, retinal pigmented epithelium, and choroid using laser scanning confocal immunofluorescence microscopy. RESULTS: Apolipoprotein E immunoreactivity is associated with photoreceptor outer segments, the retinal ganglion cell layer, the retinal pigmented epithelium basal cytoplasm and basal lamina, and with both collagenous layers of Bruch membrane. Apolipoprotein E appears to be a ubiquitous component of drusen, irrespective of clinical phenotype. It also accumulates in the cytoplasm of a subpopulation of retinal pigmented epithelial cells, many of which overlie or flank drusen. Mean levels of apolipoprotein E mRNA in the adult human retina are 45% and 150% of the levels measured in liver and adult brain, the two most abundant biosynthetic sources of apolipoprotein E. Apolipoprotein E mRNA levels are highest in the inner retina, and lowest in the outer retina where photoreceptors predominate. Significant levels of apolipoprotein E mRNA are also present in the retinal pigmented epithelium/choroid complex and in cultured human retinal pigmented epithelial cells. CONCLUSIONS: Apolipoprotein E protein is strategically located at the same anatomic locus where drusen are situated, and the retinal pigmented epithelium is the most likely local biosynthetic source of apolipoprotein E at that location. Age-related alteration of lipoprotein biosynthesis and/or processing at the level of the retinal pigmented epithelium and/or Bruch membrane may be a significant contributing factor in drusen formation and age-related macular degeneration pathogenesis. PMID- 11384576 TI - Retinopathy associated with high-dose interferon alfa-2b therapy. AB - PURPOSE: To present seven patients who developed retinopathy while receiving high dose interferon alfa-2b therapy for adjuvant treatment of high-risk melanoma. METHODS: Retrospective case series. RESULTS: Seven patients developed a retinopathy while receiving high-dose interferon alfa-2b therapy for adjuvant treatment of high-risk melanoma. Five patients had normal visual acuity, but retinopathy with cotton- wool spots and/or retinal hemorrhages with the retinopathy resolved after stopping treatment after detection. Two patients developed severe retinopathy with vision loss to counting fingers and hand motions without resolution of the retinopathy. The duration of the maintenance treatment before detection of the retinopathy for all patients ranged from 6 to 26 weeks. The total dose received at time of detection of retinopathy ranged from 816 to 1770 million units. Confounding factors included hypertension, thrombocytopenia, anemia, and a history of prior chemotherapy in one patient. Also, one patient received an investigational ganglioside vaccine, one had a history of radiation treatment to the brain, and six received paroxetine. CONCLUSIONS: Patients receiving interferon alfa-2b are at risk for developing an associated retinopathy. The risk appears to be greater with higher dosage therapy. In addition, severe vision loss can be seen with interferon alfa-2b associated retinopathy. The effect of treatment with selective serotonin reuptake inhibitors, such as paroxetine, in increasing the incidence of this complication is unknown. Patients need to be monitored until the retinopathy is resolved to screen for sequelae, including retinal neovascularization. PMID- 11384577 TI - Absence of type I estrogen receptors in choroidal melanoma: analysis of Collaborative Ocular Melanoma Study (COMS) eyes. AB - PURPOSE: To evaluate choroidal melanomas in enucleated eyes for the presence of type I estrogen receptors. METHODS: Fourteen consecutive eyes with large choroidal melanomas (defined as >16-mm basal diameter and >8 mm thickness) from 14 patients (eight women and six men with a mean age of 57 years; range, 25--74 years) enucleated in accordance with the Collaborative Ocular Melanoma Study (COMS) protocol were investigated. Immunohistochemical techniques were employed to label the choroidal melanomas for the presence of type I estrogen receptors. Each specimen was then evaluated in a masked fashion by an experienced ophthalmic pathologist for positive nuclear staining. RESULTS: No tumors showed immunohistochemical evidence of a type I estrogen receptor. CONCLUSION: Type I estrogen receptors are not present in choroidal melanoma. Estrogens are not likely to influence choroidal melanoma growth through traditional receptors. PMID- 11384578 TI - Should we use short-wavelength automated perimetry to test glaucoma patients? PMID- 11384579 TI - Pseudomonas aeruginosa in vitro corneal isolate sensitivity to ofloxacin, ciprofloxacin, and trovafloxacin: a comparative study. AB - PURPOSE: To compare sensitivity of Pseudomonas aeruginosa corneal isolates to ofloxacin, ciprofloxacin, and trovafloxacin. METHODS: Sensitivities of P. aeruginosa corneal isolates to each antibiotic from the periods 1985 to 1987 (n = 32) and 1995 to 1999 (n = 85) were evaluated in vitro with E tests (AB Biodisk; Remel, Lenexa, Kansas). RESULTS: Overall, the percent of P. aeruginosa corneal isolates sensitive in vitro to ofloxacin (106/117, 90.6%) was significantly less than to ciprofloxacin (113/117, 96.6%, P =.016) and trovafloxacin (113/117, 96.6%, P =.016). We observed trends of decreasing sensitivity to ciprofloxacin and trovafloxacin, which were not statistically significant. Sensitivity to ofloxacin remained unchanged; however, sensitivity to ofloxacin was always less than sensitivity to ciprofloxacin and trovafloxacin. CONCLUSION: Although in vitro susceptibilities may not correlate with in vivo efficacy, our data suggest that ciprofloxacin and trovafloxacin are superior to ofloxacin in the treatment of P. aeruginosa keratitis. PMID- 11384580 TI - Granulomatous kerato-conjunctivitis as a manifestation of Hodgkin lymphoma. AB - PURPOSE: To report atypical kerato-conjunctivitis as a manifestation of Hodgkin disease. METHODS: Case report. RESULTS: A 19-year old woman presented with conjunctival nodules and corneal infiltrates. Diffuse lymph node enlargement occurred, but not in the preauricular or submandibular areas. Extensive microbiologic examination was negative. Conjunctival biopsy showed noncaseating granulomas. Inguinal lymph node biopsy showed Hodgkin lymphoma, and chemotherapy was instituted. Ocular signs resolved after 6 weeks, but a corneal scar remained. CONCLUSIONS: The appearance of conjunctival granulomas and corneal infiltrates may represent a clinical manifestation of Hodgkin lymphoma. PMID- 11384581 TI - Washout periods for brimonidine 0.2% and latanoprost 0.005%. AB - PURPOSE: To evaluate the intraocular pressure washout time after discontinuing brimonidine 0.2% twice daily and latanoprost 0.005% once every evening. METHODS: We discontinued brimonidine or latanoprost in a masked fashion from primary open angle glaucoma or ocular hypertensive patients. The intraocular pressure was measured twice weekly until patients returned to untreated baseline. RESULTS: In 32 patients, the mean longest eye washout time for brimonidine (n = 15) was 3.3 +/- 3.0 weeks and for latanoprost (n = 17) was 4.4 +/- 3.2 weeks (P =.24). In all but one patient, brimonidine returned to baseline by 5 weeks and latanoprost returned by 8 weeks. CONCLUSION: After discontinuing latanoprost or brimonidine, a wide variation exists in washout times among individuals, with latanoprost demonstrating a trend to a longer washout period. PMID- 11384582 TI - Acute full-thickness macular hole after uncomplicated phacoemulsification cataract surgery. AB - PURPOSE: To report the occurrence of a full-thickness macular hole in the early postoperative period after uncomplicated phacoemulsification cataract surgery. METHODS: Retrospective analysis of a consecutive series of eyes referred for treatment of a full-thickness macular hole after uncomplicated phacoemulsification cataract surgery. RESULTS: In a 4-year period, five eyes with acute full-thickness macular hole after uncomplicated phacoemulsification cataract surgery were examined. Metamorphopsia and vision loss had occurred 2 to 8 (mean, 4.4) days after phacoemulsification. All eyes had attained normal corrected vision on day 1 postoperatively. A stage 2 full-thickness macular hole was present in four of the five, and a stage 3 full-thickness macular hole in one of the five eyes with acuities of 20/60--20/120 (median, 20/80). All five eyes had successful closure with early primary full-thickness macular hole surgery with visual improvement to 20/20-- 20/60 (median, 20/40). CONCLUSION: Full thickness macular hole may occur rarely during the early postoperative period after uncomplicated phacoemulsification, and early diagnosis and full-thickness macular hole surgery carries a favorable prognosis. The mechanisms underlying macular hole formation in these eyes are unclear. PMID- 11384583 TI - Retinal vasculitis and posterior pole "hypopyons" as early signs of acute bacterial endophthalmitis. AB - PURPOSE: To report a case of acute postoperative bacterial endophthalmitis presenting with retinal vasculitis and posterior pole "hypopyons." DESIGN: Observational case report. METHODS: Observational case report. RESULTS: In a 73 year-old woman, acute postoperative endophthalmitis presented with retinal vasculitis, diffuse retinal hemorrhages, and several posterior pole "hypopyons" that were collections of yellow-white inflammatory debris. The inflammation was located primarily in the posterior one third of the vitreous, and except for severely impaired visual acuity, the classic signs of acute postoperative endophthalmitis were not present until the next day. An immediate pars plana vitrectomy with intravitreal injection of vancomycin, amikacin, and dexamethasone was performed. Vitreous and aqueous cultures grew coagulase-negative Staphylococcus species. After intravenous and topical therapy for endophthalmitis, the inflammation subsided. Eight weeks after the initial cataract surgery, the best corrected visual acuity of the patient was 20/25. CONCLUSION: Acute postoperative bacterial endophthalmitis may present with atypical clinical signs and inflammation located primarily in the retina and posterior vitreous. PMID- 11384584 TI - Cryoglobulinemia associated with Purtscher-like retinopathy. AB - PURPOSE: To report a case of Purtscher-like retinopathy in a young man with chronic hepatitis C--associated cryoglobulinemia. METHODS: Case report. RESULTS: A 44-year-old Caucasian man with chronic hepatitis C developed sudden loss of vision in the left eye and abdominal pain. Ocular fundus examination revealed peripapillary cotton-wool spots and superficial retinal whitening in the macula. Systemic evaluation revealed markedly elevated rheumatoid factor, hypertension, and acute renal insufficiency. A renal biopsy demonstrated intravascular deposition of immunoglobulins IgG and IgM and complement consistent with type II mixed cryoglobulinemia. CONCLUSION: Clinicians should consider cryoglobulinemia in the differential diagnosis of a patient with Purtscher-like retinopathy and history of infection with hepatitis C. PMID- 11384585 TI - Central serous chorioretinopathy after bone marrow transplantation. AB - PURPOSE: To describe central serous chorioretinopathy after bone marrow transplantation. METHODS: The medical records of the patient were reviewed retrospectively. RESULTS: A 46-year-old Filipino man developed multifocal central serous chorioretinopathy affecting his left eye 4 months after bone marrow transplantation for acute myelogenous leukemia. Other co-existing medical problems at the time of presentation included systemic hypertension and graft versus-host-disease (GVHD), for which the patient was using both systemic corticosteroids and cyclosporine. CONCLUSION: Central serous chorioretinopathy is a rare cause of vision loss in patients after bone marrow transplantation. Previous descriptions of bone marrow transplantation-associated central serous chorioretinopathy in patients with thrombotic microangiopathy, as well as the occurrence of both systemic hypertension and the use of systemic corticosteroids and cyclosporine in our patient with bone marrow transplantation-associated central serous chorioretinopathy, support theories of choroidal vascular compromise in the pathogenesis of central serous chorioretinopathy. PMID- 11384586 TI - Retinal anti-bipolar cell antibodies in a patient with paraneoplastic retinopathy and colon carcinoma. AB - PURPOSE: To describe clinical, electrophysiologic, and immunologic features of a unique paraneoplastic retinopathy with characteristics of cancer-associated and melanoma-associated retinopathy. METHODS: Serial assessment of clinical visual function, electroretinography, and assays of anti-retinal antibodies. RESULTS: A 51-year-old woman with progressive visual glare for 1 year had normal visual acuity and color vision, paracentral scotomas, and a normal-appearing retina. Electroretinography revealed no responses of the right eye and attenuated responses of the left eye, especially those recorded under scotopic conditions. Anti-bipolar antibodies were detected. Subsequent evaluation uncovered adenocarcinoma of the colon. Several months after resection of the tumor and chemotherapy, no evidence existed of cancer or anti-bipolar cell antibodies, and electroretinography responses were markedly improved. CONCLUSION: The presence of anti-bipolar cell antibodies in a patient with retinal dysfunction is not specific of melanoma-associated retinopathy. Effective treatment of cancer may result in elimination of associated anti-retinal antibodies and improved retinal function. PMID- 11384587 TI - Retinopathy associated with Machado--Joseph disease (spinocerebellar ataxia 3) with CAG trinucleotide repeat expansion. AB - PURPOSE: To report characteristic atrophic maculopathy in a patient with Machado- Joseph disease (spinocerebellar ataxia 3) caused by trinucleotide repeat expansion of the relevant gene. METHODS: Case report. RESULTS: A 64-year-old Japanese man had suffered from slurred speech and gait disturbance since 57 years of age. Cerebellar ataxia, extensor plantar response, and other neurological signs were compatible with features of Machado--Joseph disease. Magnetic resonance imaging showed atrophies of cerebellum and cerebral cortex. Family history suggested an autosomal dominant inheritance of the disease. The patient presented with gaze-evoked nystagmus and limitations of eye movement in all directions. Ophthalmoscopy and fluorescein angiogram revealed symmetric changes in the posterior fundi, which consisted of patchy atrophies at the level of the retinal pigment epithelium. Scotopic electroretinogram showed no abnormalities with normal oscillatory potentials. Polymerase chain reaction analysis of the Machado--Joseph disease gene identified a heterozygous trinucleotide (CAG) repeat expansion. CONCLUSION: This case illustrates a rare association of atrophic maculopathy and external ophthalmoplegia in Machado--Joseph disease, contrasted with the common occurrence of retinal degeneration in spinocerebellar ataxia 7. Dystrophic changes in the retinal pigment epithelium have rarely been described but may be one of the characteristic complications of Machado--Joseph disease. PMID- 11384588 TI - Dominant radial drusen and Arg345Trp EFEMP1 mutation. AB - PURPOSE: To report a new North American family with dominant radial drusen and Arg345Trp mutation in the EFEMP1 gene. METHODS: Clinical and molecular genetic family study. RESULTS: Four family members had macular drusen, and one had submacular fibrosis and visual loss. An Arg345Trp mutation of the EFEMP1 gene was detected in three affected family members, but not in three unaffected members. CONCLUSION: The Arg345Trp mutation remains the only cause of Doyne hereditary macular dystrophy, also known as Malattia Leventinese or radial dominant drusen. PMID- 11384589 TI - Axial length and refractive error in X-linked retinoschisis. AB - PURPOSE: To examine the relationship between axial length and refractive error in patients with X-linked retinoschisis. DESIGN: To determine whether the hypermetropia frequently found in patients with X-linked retinoschisis is axial hypermetropia. METHODS: The axial length and refractive error were measured in 29 right eyes of 29 patients. The patients were divided into two groups: a juvenile group with ages <13 years (12 eyes) and an adult group with ages > or =13 years (17 eyes). The axial length of the right eye of 30 adult men without eye diseases whose refractive error ranged from +/- 1.0 diopter served as controls. RESULTS: In the adult patient group, the refractive error was significantly more hypermetropic and the axial length was significantly shorter than was the normal adult group (P <.001). CONCLUSION: These results strongly suggest that the hypermetropia in patients with X-linked retinoschisis is axial hypermetropia. PMID- 11384590 TI - Evaluation of retinal damage induced by air/fluid exchange using a trypan blue inclusion test in rabbits. AB - PURPOSE: To investigate whether retina damaged in the process of air/fluid exchange could be visualized in situ using trypan blue staining. METHODS: Vitrectomy and air/fluid exchange with continuous air infusion for various periods of time was performed in adult pigmented rabbits, followed by injection of 0.4% trypan blue solution into the vitreous cavity. Retinal staining was evaluated intraoperatively under the operating microscope and by light microscopy after enucleation. RESULTS: Intraoperative examination showed localized trypan blue staining of areas of the retina opposite the infusion cannula. Light microscopy of the same areas revealed intranuclear trypan blue inclusion in the retina. CONCLUSION: Trypan blue staining allows for the immediate in situ evaluation of damage to the retina associated with air/fluid exchange and continuous air infusion in rabbit eyes. PMID- 11384591 TI - Alternating ptosis after bilateral frontalis muscle suspension for congenital ptosis. AB - PURPOSE: To report an unusual response to bilateral frontalis suspension surgery for congenital ptosis. METHODS: Case report. RESULTS: A 6-year-old child with infantile esotropia and bilateral congenital ptosis received a homologous fascia lata frontalis suspension. After surgery, the patient exhibited an alternating ptosis, caused by alternating unilateral frontalis muscle contraction, which has persisted throughout 3 years of postoperative observation. CONCLUSION: Symmetric frontalis muscle activation may not always occur after bilateral frontalis muscle suspension surgery for congenital ptosis. Amblyopia, strabismus, or fixation preference may predispose patients to use unilateral brow elevation, despite bilateral frontalis muscle suspension surgery. PMID- 11384592 TI - Bilateral abducens nerve paresis associated with anti-GQ1b IgG antibody. AB - PURPOSE: To describe two cases with bilateral abducens nerve paresis associated with serum anti-GQ1b IgG antibody. METHODS: Case reports. RESULTS: Two boys, aged 12 and 10 years, experienced acute onset of diplopia after preceding symptoms and signs of infection. In both boys, examination showed only bilateral abducens nerve paresis. Although routine laboratory data and magnetic resonance imaging demonstrated no pathologic findings, titer of anti-GQ1b IgG antibody in the sera of both patients was increased. Diplopia and signs of bilateral abducens nerve paresis disappeared in 6 weeks without any specific treatment. CONCLUSION: The anti-GQ1b IgG antibody in the sera of both patients probably contributed to the bilateral abducens nerve paresis. PMID- 11384593 TI - Delayed orbital hemorrhage after routine strabismus surgery. AB - PURPOSE: To report a case of delayed rectus muscle hemorrhage after strabismus surgery. METHODS: Case report. RESULTS: Rectus muscle hemorrhage occurred 36 hours after strabismus surgery in a 26-year-old man, causing temporary loss of vision and reduced ocular motility. Urgent lateral cantholysis and orbital exploration to restore hemostasis were undertaken. Full recovery of vision occurred and a small residual motility disturbance was present 3 months postoperatively. CONCLUSION: Delayed rectus muscle hemorrhage poststrabismus surgery is rare but can have sight-threatening effects. When vision is threatened because of optic nerve compromise, urgent orbital exploration may allow full recovery of function. PMID- 11384606 TI - Antagonism of CRF(2) receptors produces anxiolytic behavior in animal models of anxiety. AB - Two pharmacologically distinct CRF receptors are distributed in different brain regions and peripheral tissues. Studies suggest that CRF(1) receptors play an important role in mediating the anxiety provoking effects of CRF. In contrast, far less functional information is available on CRF(2) receptors. Therefore, we conducted dose response studies using antisauvagine-30 (anti-SVG-30, 0-20 microg, 20-min pretreatment, i.c.v.), a potent CRF(2) peptide antagonist, and tested rats in three models of anxiety - the conditioned freezing, the elevated plus maze, and the defensive-withdrawal test. Anti-SVG-30 produced a significant dose dependent reduction in conditioned freezing. In the elevated plus maze test, administration of anti-SVG-30 effectively increased the number of entries and time spent in the open arms. In the defensive-withdrawal test, anti-SVG-30 treatment facilitated exploratory activity in a large illuminated open field. Thus, in all three animal models, administration of anti-SVG-30 was consistent in producing an anxiolytic-like behavioral effect. In addition, a dose of anti-SVG 30 (10 microg) that produced anxiolytic-like behavior had no significant effects on locomotor activity measured in an automated activity box. This latter finding suggests that antagonism of CRF(2) receptors is not associated with a non specific increase in behavioral movements. These results provide evidence that, in addition to CRF(1) receptors, CRF(2) receptors may play an important role in the mediation of anxiety behavior. PMID- 11384607 TI - Differential localization and colocalization of two neuron-types of sodium dependent inorganic phosphate cotransporters in rat forebrain. AB - We studied by immunohistochemistry the distribution of differentiation-associated sodium-dependent inorganic phosphate (Pi) cotransporter (DNPI) in the rat forebrain, in comparison with brain-specific cotransporter (BNPI). DNPI-staining was principally seen in axonal synaptic terminals which showed a widespread but discrete pattern of distribution different from that of the BNPI-staining. In the diencephalon, marked DNPI-staining was seen in the dorsal lateral geniculate, medial geniculate, ventral posterolateral, ventral posteromedial, anterior, and reticular thalamic nuclei without the colocalization with BNPI-staining. DNPI staining showed a strong mosaical pattern and overlapped well the BNPI-staining in the medial habenular nucleus. DNPI-staining was moderate over the hypothalamus and notably localized in neurosecretory terminals containing corticotropin releasing hormone in the median eminence. In contrast, the BNPI-staining was region-related and strong in the ventromedial and mammillary nuclei. In the telencephalon, laminar DNPI-staining was seen over the neocortex, corresponding to the thalamocortical termination, and also found in the retrosplenial cortex and the striatum, with the highest intensity in the accumbens nucleus shell. The present results suggest that DNPI serves as a dominant Pi transport system in synaptic terminals of diencephalic neurons including thalamocortical and thalamostriatal pathways as well as the hypothalamic neuroendocrine system in the rat forebrain. PMID- 11384608 TI - The uptake of cysteine in cultured primary astrocytes and neurons. AB - One of the vitally important functions of glutathione (GSH) is to adequately protect cells against toxic chemicals, reactive oxygen metabolites and free radical species. The amino acid, cysteine, is the key rate-limiting substrate for the biosynthesis of GSH, and the maintenance of adequate intracellular GSH levels is dependent upon the extracellular availability and transport of cysteine into cells. In the present study, primary cultures of astrocytes and neurons were employed to characterize cysteine transport systems. Both astrocytes and neurons used Na(+)-dependent systems as the major route for cysteine uptake (80-90% of total), while Na(+)-independent uptake represented a minor component of total transport (10-20% of total). Among the Na(+)-dependent systems, X(AG(-)) was the major contributor (approx. 80-90%) for cysteine uptake in both neurons and astrocytes, with a minor contribution from the ASC transport system (Na(+) dependent neutral amino acid transport system for alanine, serine, and cysteine). In the Na(+)-independent transport systems (10-20% of total cysteine transport), multifunctional ectoenzyme/amino acid transporter gamma-glutamyltranspeptidase (GGT), and the neutral amino acid L-system contributed approximately equally towards cysteine uptake, in both neurons and astrocytes. The present studies demonstrate that astrocytes and neurons accumulate cysteine by both Na(+) dependent and Na(+)-independent uptake systems, with major uptake occurring through the X(AG(-)) system and minor uptake via the ASC, GGT and L-systems. PMID- 11384609 TI - Meal-induced changes in extracellular 5-HT in medial hypothalamus of lean (Fa/Fa) and obese (fa/fa) Zucker rats. AB - Hypothalamic serotonin (5-HT) is involved in appetite regulation and sympathetic stimulation of thermogenesis. This study tested the hypothesis that the enhanced energetic efficiency of obese Zucker rats involves blunted serotonergic release within the medial hypothalamus (MH). We used microdialysis and HPLC-EC to measure dynamic changes in extracellular 5-HT levels in the MH of 10-13-week-old male lean (Fa/Fa) and obese (fa/fa) Zucker rats before and after a meal. No differences were noted in basal levels of 5-HT between lean and obese rats. Consistent with the suggestion that hypothalamic 5-HT plays a physiological role in feeding, extracellular 5-HT levels increased significantly in both lean and obese rats given a meal. This increase was observed in the 20 min interval in which they ate the 8.1 kcal meal and remained for an additional 60 min. The net release of 5-HT during the meal interval was comparable in the lean (1.46+/-0.38 fmol/microl) and obese (1.21+/-0.82 fmol/microl) rats. However, the 5-HT levels of the leans (1.80+/-0.29 fmol/microl) plateaued in the next 20 min interval, whereas they continued rising (2.74+/-0.53 fmol/microl) in obese rats and were significantly higher than those in the leans during the 40 and 60 min intervals after the meal was presented. This resulted in a total net release during the meal plus the next three 20 min intervals that was significantly higher in obese (9.83+/-1.16 fmol/microl) than in lean (5.59+/-0.85 fmol/microl) rats. Thus, the enhanced energetic efficiency of the obese Zucker rats may not be associated with attenuated serotonin release in response to a meal. Rather their enhanced release of 5-HT in the MH may reflect compensatory mechanisms for the elevated orexigen NPY, the reduction in meal-induced CCK release, and/or a functional resistance to 5-HT. PMID- 11384610 TI - Monocyte chemoattractant protein-1 expressed in neurons and astrocytes during focal ischemia in mice. AB - Focal cerebral ischemia elicits an inflammatory response characterized by the infiltration and accumulation of leukocytes, as well as the secretion of inflammatory mediators (Clark et al., Brain Res. Bull., 35 (1994) 387-392; Garcia et al., Am. J. Pathol., 144 (1994) 188-199; Wang et al., J. Neurochem. 71 (1998) 1194-1204). Leukocytes eliminate microbial invaders and necrotizing tissue debris, and can also turn against surrounding healthy tissue and exacerbate tissue injury (Furie and Randolph, Am. J. Pathol., 146 (1995) 1287-1301; Kochanek and Hallenbeck, Stroke 23 (1992) 1367-1379). Inflammatory mediators are considered to play an important role in attracting and stimulating leukocytes (Weiss, N. Engl. J. Med., 320 (1989) 365-376). Monocyte chemoattractant protein-1 (MCP-1) functions as an inflammatory mediator, whose source and role in focal cerebral ischemia is worth studying. MCP-1, a potent chemoattractant factor, may play an important role in ischemia-induced inflammatory response. The aim of the present study is to determine the time course and cell type of MCP-1 protein expression after permanent focal ischemia in mice. ELISA and immunohistochemical staining were used to detect the expression of MCP-1 protein after 0 h, 2 h, 4 h, 12 h, 1 day, 2 days, 3 days, 5 days and 7 days of middle cerebral artery occlusion (n=3-5 in each group). Double-labeled fluorescent staining was used to examine the cellular localization of MCP-1. The results demonstrated that MCP-1 expression was mainly observed in the ischemic core after 12 h of middle cerebral artery occlusion, then gradually increased and extended to the ischemic perifocal area. MCP-1 expression peaked at 2 days and 3 days, and gradually decreased after 5 days of MCAO. Double-labeled immunostaining for MCP-1 and neuron specific enolase (NSE) or glial fibrillary acidic protein (GFAP) showed that MCP-1 positive neurons were observed as early as 12 h of ischemia, while MCP-1 positive astrocytes were observed after 2 days of ischemia. These results support the functional role of MCP-1 in ischemic brain injury and reveal a distinct temporal and spatial expression of MCP-1 in cells believed to be neurons and astrocytes. PMID- 11384611 TI - Time course of glial proliferation and glial apoptosis following excitotoxic CNS injury. AB - Activation of microglial cells and astrocytes after CNS injury results in changes in their morphology, immunophenotype and proliferative activity and has neurotrophic as well as neurotoxic consequences. However, little is known about the exact time course of glial activation as regards their proliferative activity and their fate. In this study, quantification of the densities of proliferating and non-proliferating microglial cells and astrocytes was carried out over 30 days by counting differentially labeled cells in the striatum and substantia nigra pars reticulata (SNr) after injection of quinolinic acid into the rat striatum. The TdT-mediated dUTP nick end labeling (TUNEL)-reaction was used to detect possible apoptotic mechanisms which limit the glial reaction. At 1 day post injection (p.i.) non-proliferating ameboid microglia/macrophages were seen in the striatum, but at 3 and 5 days p.i. many proliferating, ameboid microglia/macrophages and hypertrophic microglia were detected. At 10 days p.i., the time point with the highest density of hypertrophic microglia, TUNEL-positive microglial cells were observed indicating that apoptotic processes play a role in restricting this reaction. In contrast to this, at early time points, a reduction in the density and glial fibrillary acidic protein (GFAP)-immunoreactivity of astrocytes in the striatum was detected. At later time points, a dense astrogliosis with proliferating astrocytes developed in the dorsal and medial striatum. At 30 days p.i., in the entire striatum a dense astrogliosis was detected. The SNr showed a short period of microglial activation and proliferation and a long lasting astrogliosis without proliferation PMID- 11384612 TI - Characterization of cytosolic glutathione S-transferase in cultured astrocytes. AB - To elucidate the contribution of glutathione S-transferase (GST) and glutathione peroxidase (GPx) to the protection against oxidative stress in rat brain, we prepared GST and GPx from newborn rat liver, brain and cultured astrocytes, and investigated the characteristics and kinetics of the enzymes. The activity of cytosolic GST of the cultured astrocytes toward 1-chloro-2,4-dinitrobenzene (CDNB) was much higher than that of GPx toward peroxides. The GST activity toward 4-hydroxy-2-nonenal (4HNE) was almost the same as the GPx activity. GST isozymes were purified from the cytosolic fraction of the liver and astrocytes. In the case of the astrocytes, a major GST isozyme with an isoelectric point (pI) of 9.02 accounted for approximately 40% of total GST activity toward CDNB, while hepatic GST isozymes showed seven peaks in the basic region. Each of astrocytes and liver showed a single GST peak with high activity toward 4HNE, namely AVIII and LVIII, respectively, and both of them had a similar pI value of about 6.7. The kinetic parameters of AVIII and LVIII were found to be similar to each other. These data suggest that the same types of GST isozymes are expressed in the astrocytes and liver, and take part mainly in the detoxification of 4HNE. PMID- 11384613 TI - Immunostimulation of rat primary astrocytes decreases intracellular ATP level. AB - In this study we investigated the effect of immunostimulation on intracellular ATP level in rat glial cells. Rat primary astrocytes or C6 glioma cells were treated for 48 h with IFN-gamma, LPS or IFN-gamma plus LPS. These treatments increased NO production from the cells and a synergistic increase in NO production was observed with IFN-gamma plus LPS. Intracellular ATP level was decreased to about half the control level at the highest concentration of IFN gamma (100 U/ml) plus LPS (1 microg/ml) without affecting cell viability. The level of intracellular ATP was inversely correlated with the extent of NO production from the glial cells. The increase in NO production is at least 6 h ahead of the initiation of ATP depletion, and NOS inhibitor N(G)-nitro-L-arginine (NNA) or Nomega-nitro-L-arginine methyl ester (L-NAME) inhibited NO production and ATP depletion. Exogenous addition of peroxynitrite generator 3 morpholinosydnonimine (SIN-1) and to a lesser extent NO generator S-nitroso-N acetylpenicillamine (SNAP) depleted intracellular ATP level in a dose-dependent manner. The results from the present study imply that immunostimulation of rat glial cells decreases the intracellular ATP level without affecting cell viability. Considering the role of astrocytes as an essential regulator of the extracellular environment in the brain, the immunostimulation-induced decrease in intracellular ATP level may participate in the pathogenesis of various neurological diseases. PMID- 11384614 TI - Identification of dual specificity phosphatases induced by olfactory bulbectomy in rat olfactory neuroepithelium. AB - Dual specificity protein tyrosine phosphatases (dsPTPs) are a subfamily of protein tyrosine phosphatases implicated in the regulation of extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK), and p38 mitogen-activated protein kinases (MAPKs) which are target enzymes activated by a wide range of cell-surface stimuli. Like these kinases, a class of dsPTP has been implicated in cell differentiation, regeneration, and apoptosis. In order to isolate dsPTPs which might play an important role in neuronal regeneration and apoptosis in olfactory neuroepithelium, we subcloned DNA fragments amplified by reverse transcription polymerase chain reaction (RT-PCR), using degenerate oligonucleotide primers based on the conserved amino acid regions within the catalytic domain of dsPTPs, from rat olfactory epithelial RNA 1 and 4 h after an olfactory bulbectomy. The PCR products were subcloned into the pCRII vector, and 23 clones were chosen for further characterization. The sequence of these 23 individual clones revealed that two clones were identical to the rat dsPTP, MKP-3, and the other 21 clones were identical to the rat dsPTP, MKP-1. By Northern analysis, the MKP-1 transcript was induced and peaked 4 h following a bulbectomy. Similar results were obtained with the MKP-3 transcript. These results suggest that MKP-1 and MKP 3 may be involved in the early steps of apoptosis in vivo in rat olfactory neuroepithelium. PMID- 11384615 TI - Arginine-vasopressin in nucleus of the tractus solitarius induces hyperglycemia and brain glucose retention. AB - Hypothalamic arginine-vasopressin (AVP) plays an important role both as a neurotransmitter and hormone in the regulation of blood glucose and feeding behavior. AVP-containing axons from the parvocellular subdivision of paraventricular nucleus of the hypothalamus terminate in the nucleus of the tractus solitarius (NTS), but the function of this projection is not known. Interestingly, the NTS also receives afferent information from the carotid body and other peripheral receptors involved in glucose homeostasis. We have previously reported that stimulation of the carotid body receptors initiates a hyperglycemic reflex and increases brain glucose retention. Here we show that direct administration of micro-doses of AVP into the NTS of anesthetized or awake rats rapidly increased the levels of blood glucose concentration and brain arterio-venous (A-V) glucose difference. This effect was not observed when the same doses of AVP were injected in the brainstem outside NTS. Arginine vasopressin antagonist microinjections alone produced a small but significant reduction in brain A-V glucose. Pre-administered VP1-receptor antagonist [beta mercapto-beta,beta-cyclopentamethylene-propionyl(1),O-Me Tyr(2),Arg(8)]vasopressin blocked the effects of AVP. These results indicate that AVP acting on its receptors locally within the NTS participates in glucose homeostasis, increasing both blood glucose concentration and brain A-V glucose differences. Hypothalamic AVP may facilitate hyperglycemic responses initiated by peripheral signals processed at the level of the NTS. PMID- 11384616 TI - Nitric oxide enhances MPP(+)-induced hydroxyl radical generation via depolarization activated nitric oxide synthase in rat striatum. AB - We examined the effect of N(G)-nitro-L-arginine methyl ester (L-NAME), a nitric oxide synthase (NOS) inhibitor, on extracellular potassium ion concentration ([K(+)](o))-enhanced hydroxyl radical (.OH) generation due to 1-methyl-4 phenylpyridinium ion (MPP(+)) was examined in the rat striatum. Rats were anesthetized, and sodium salicylate in Ringer's solution (0.5 nmol/microl per min) was infused through a microdialysis probe to detect the generation of.OH as reflected by the non-enzymatic formation of 2,3-dihydroxybenzoic acid (DHBA) in the striatum. Induction of KCl (20, 70 and 140 mM) increased MPP(+)-induced.OH formation trapped as 2,3-dihydroxybenzoic acid (DHBA) in a concentration dependent manner. However, the application of L-NAME (5 mg/kg i.v.) abolished the [K(+)](o) depolarization-induced.OH formation with MPP(+). Dopamine (DA; 10 microM) also increased the levels of DHBA due to MPP(+). However, the effect of DA after application of L-NAME did not change the levels of DHBA. On the other hand, the application of allopurinol (20 mg/kg i.v., 30 min prior to study), a xanthine oxidase (XO) inhibitor was abolished the both [K(+)](o)- and DA induced.OH generation. Moreover, when iron(II) was administered to MPP(+) then [K(+)](o) (70 mM)-pretreated animals, a marked increase in the level of DHBA. However, when corresponding experiments were performed with L-NAME-pretreated animals, the same results were obtained. Therefore, NOS activation may be no relation to Fenton-type reaction via [K(+)](o) depolarization-induced.OH generation. The present results suggest that [K(+)](o)-induced depolarization augmented MPP(+)-induced.OH formation by enhancing NO synthesis. PMID- 11384617 TI - Persistent cholinergic presynaptic deficits after neonatal chlorpyrifos exposure. AB - The commonly-used organophosphate insecticide, chlorpyrifos (CPF), impairs brain cell development, axonogenesis and synaptogenesis. In the current study, we administered CPF to neonatal rats on postnatal (PN) days 1-4 (1 mg/kg) or PN11-14 (5 mg/kg), treatments that were devoid of overt toxicity. We then examined two cholinergic synaptic markers, choline acetyltransferase activity (ChAT) and [3H]hemicholinium-3 binding (HC-3) in the hippocampus, midbrain, striatum, brainstem and cerebral cortex in the juvenile (PN30) and young adult (PN60). Across all brain regions, CPF exposure evoked significant reductions in both markers, with larger effects on HC-3 binding, which is responsive to neuronal impulse activity, than on ChAT, a constitutive marker. Superimposed on the deficits, there were gender-selective effects and distinct regional disparities in the critical exposure period for vulnerability. In the hippocampus, either the early or late treatment regimen evoked decreases in ChAT but the early regimen elicited a much larger decrease in HC-3; effects persisted into adulthood. In the midbrain, CPF administration on PN1-4 elicited deficits similar to those seen in the hippocampus; however, exposure on PN11-14 elicited changes preferentially in females. Gender selectivity was also apparent in the striatum, in this case reflecting deficits in females after CPF treatment on PN1-4. In contrast, the effects of CPF on the brainstem were relatively more robust in males; effects in the cerebral cortex were less notable than in other regions. These results indicate that neonatal CPF exposure produces widespread deficiencies in cholinergic synaptic function that persist into adulthood. The effects are likely to contribute to gender-selective alterations in behavioral performance that persist or emerge long after the termination of exposure and well after the restoration of cholinesterase activity. PMID- 11384618 TI - An analysis of the effect of retinal ganglion cell impulses upon the firing probability of neurons in the dorsal lateral geniculate nucleus of the cat. AB - This study examines the probabilistic way in which LGN cells produce impulses. Simultaneous extracellular recordings were made from a single lateral geniculate nucleus (LGN) relay cell and the one retinal cell that triggered most of its impulses during vigorous responses. Impulses in the retinal train were classified as 'successful' in triggering an LGN impulse, or 'unsuccessful'. Similarly, the impulses in the LGN train were either 'Triggered' by a successful retinal impulse, or were 'Anonymous'. These impulses delimited various intervals whose distributions were compared to the relevant distribution of all intervals to determine whether short or long intervals tended to dominate in each case. Intervals between unsuccessful and successful impulses tended to be shorter than other retinal intervals, with their probability declining exponentially with duration. These data imply a decaying excitation produced by each impulse, but with a short refractory period following each Triggered impulse. Short intervals between Anonymous impulses were relatively common; Anonymous impulses thus lack the same refractoriness and tend to occur in bursts. The exponential excitation following an unsuccessful retinal impulse also facilitates Anonymous impulses, while Anonymous impulses (during visual stimulation) render the LGN slightly refractory for subsequent retinal impulses. PMID- 11384619 TI - Effects of soft-diet feeding on synaptic density in the hippocampus and parietal cortex of senescence-accelerated mice. AB - Some investigators have proposed that extracting of the teeth of rats or mice impairs their acquisition of spatial memory, implying that alterations of the neural networks in the brain result from a reduction of masticatory work. To evaluate numerical alterations of synapses in the cerebral cortex caused by reduced masticatory movements, two strains of the senescence-accelerated mouse, SAMR1 and SAMP8, were fed either a pelleted (hard-diet groups, R1-H and P8-H) or a powdered diet (soft-diet groups, R1-S and P8-S) after weaning. Radioimmunoassay using a monoclonal anti-synaptophysin antibody (SY38) revealed that the synaptophysin content in the whole cortex was significantly lower in P8-H compared with R1-H from 3 months to 12 months of age. The soft-diet feeding reduced the synaptophysin content in the cerebral cortex of both strains after 3 months of age. Immunohistochemistry and electron microscopy on the hippocampal formation and parietal cortex of 6-month-old mice showed that synaptic formation was significantly decreased in these areas in both R1-S and P8-S. The reduction rate of synaptic density due to soft-diet feeding was larger in the hippocampus than in the parietal cortex. The working memory of the four groups was tested at 6 months of age on an eight-arm radial maze. Performance significantly differed between R1-H and P8-H, between R1-H and R1-S, and between P8-H and P8-S. The results indicated that soft-diet feeding after weaning period reduces synaptic formation in the cerebral cortex and impairs the ability of spatial learning in adulthood. PMID- 11384620 TI - Thrombin inhibition attenuates neurodegeneration and cerebral edema formation following transient forebrain ischemia. AB - The disturbance of microcirculation following cerebral ischemia leads to an enlargement of cerebral infarct volume. Endogenous thrombin may play a role in this disturbance of microcirculation following cerebral ischemia. Therefore, the inhibition of thrombin may improve neurodegeneration and the accumulation of cerebral edema following cerebral ischemia in gerbils. The effects of thrombin inhibitor (argatroban) on cerebral ischemia were investigated in comparison with thromboxane A2 synthase inhibitor (ozagrel) and cyclooxygenase inhibitor (aspirin) following bilateral common carotid artery occlusion and reperfusion (CCA:O/R) in male Mongolian gerbils. This study consisted of three experiments: (1) morbidity and survival ratio (n=40 for each), (2) histopathology (n=12 for each), and (3) mean arterial blood pressure, local cerebral blood flow (CBF), and cerebral specific gravity (n=8 for each). Argatroban treatment improved survival ratio and stroke index, and decreased ischemically injured cell numbers in cortex and hippocampus and cerebral edema in cortex compared with aspirin and saline, in concert with the fast recovery of local CBF without reactive hyperemia following bilateral CCA:O/R. Ozagrel treatment also improved those factors compared with saline, in concert with the fast recovery of local CBF with reactive hyperemia. Aspirin treatment improved survival ratio and stroke index, and decreased ischemically injured cell numbers in cortex. Thrombin inhibition with argatroban decreases neurodegeneration and cerebral edema following bilateral CCA:O/R in gerbils. PMID- 11384621 TI - Glial cell line-derived neurotrophic factor enhances axonal regeneration following sciatic nerve transection in adult rats. AB - Adult rat sciatic nerve was transected and sutured with an entubulation technique. The nerve interstump gap was filled with either collagen gel (COL) or collagen gel mixed with glial cell line-derived neurotrophic factor (COL/GDNF). Four weeks after nerve transection, horseradish peroxidase (HRP)-labelled spinal cord motoneurons and the myelinated distal stump axons were quantified. Compared with the COL group, the percentages of labeled spinal somas and axon number were significantly increased after topically applied glial cell line-derived neurotrophic factor (GDNF). The functional recovery of the transected nerve was improved in COL/GDNF group. GAP-43 expression was also significantly higher in COL/GDNF group 1 and 2 weeks after sciatic nerve axotomy vs. COL group. These data provide strong evidence that GDNF could promote axonal regeneration in adult rats, suggesting the potential use of GDNF in therapeutic approaches to peripheral nerve injury and neuropathies. PMID- 11384622 TI - Immunohistochemical localization of neprilysin in the human cerebral cortex: inverse association with vulnerability to amyloid beta-protein (Abeta) deposition. AB - We investigated the immunohistochemical localization of neprilysin, a putative amyloid beta-protein (Abeta)-degrading enzyme, in postmortem human brain tissues. In the cerebral cortex, neprilysin immunoreactivity was weak, but relatively dense distribution was found in the primary somatosensory and visual cortices compared with the hippocampus and association cortices. In Alzheimer brain, neprilysin-positive dystrophic neurites occurred in senile plaques in the primary cortices, an observation that supports the relative abundance of neprilysin positive neuronal processes. A paucity of neprilysin in the hippocampus and association cortices may contribute to the vulnerability of these areas to Abeta deposition. PMID- 11384623 TI - Peritoneal implantation of macroencapsulated porcine pancreatic islets in diabetic rats ameliorates severe hyperglycemia and prevents retraction and simplification of hippocampal dendrites. AB - The hippocampus of rats with uncontrolled insulin-dependent diabetes undergoes retraction and simplification of apical dendrites of the CA3 pyramidal neurons and synaptic rearrangements within mossy fiber terminals that could alter hippocampal connectivity and function. The intraperitoneal implantation of hydrophilic agarose macrobeads containing porcine islets for 17 days in rats with streptozotocin-induced diabetes results in normalization of body weight gain, significant control of hyperglycemia and prevention of hippocampal dendritic remodeling, and therefore, provides an effective therapeutic option. PMID- 11384624 TI - Induction of highly polysialylated neural cell adhesion molecule (PSA-NCAM) in postischemic gerbil hippocampus mainly dissociated with neural stem cell proliferation. AB - We investigated a possible expression of highly polysialylated neural cell adhesion molecule (PSA-NCAM) in gerbil hippocampus after 5 min of transient global ischemia in association to the proliferation of neural stem cell labeled with bromodeoxyuridine (BrdU). The number of PSA-NCAM positive cells increased in the granule cell layer (GCL) of dentate gyrus (DG) by 1.9 to 2.7-fold at 10 and 20 days after the reperfusion. The number of BrdU-labeled cells increased mainly in the subgranular zone of DG by 7.2 to 8.0-fold at 5 and 10 days after the reperfusion. Immunofluorescence for PSA-NCAM and BrdU showed that the majority of DG cells were not double labeled, while one or two cells per section were double labeled in the deepest portion of the GCL only at 10 days after the reperfusion. These results suggest different predominant spatial distribution and chronological change of PSA-NCAM positive and BrdU-labeled cells in DG after transient ischemia. PMID- 11384626 TI - The future of human ovarian cryopreservation and transplantation: fertility and beyond. AB - OBJECTIVE: To review the current progress in ovarian cryopreservation and transplantation and to discuss the obstacles with the clinical application of this technique. DESIGN: The literature on ovarian cryopreservation and transplantation was reviewed to facilitate understanding and predict future directions. The studies related to this topic were identified through MEDLINE and other bibliographic databases, focusing on the most recent developments. CONCLUSION(S): The experimental evidence for low-temperature storage of ovarian tissue is encouraging. Although restoration of fertility with cryopreserved ovarian grafts has been successful in various animals, there are uncertainties about the optimum use of stored ovarian tissue in humans. Autotransplantation appears to be promising, but the potential risk of transmitting malignant cells in women with cancer is of great concern. The maturation of primordial follicles with xenotransplantation is encouraging, but the efficacy and the safety of this method need further investigation. Furthermore, the quality of oocytes that have been matured in a host animal is unknown. The development of in vitro culture systems for oocyte maturation is still in its infancy. There are many issues to be resolved in ovarian transplantation before the full clinical use of this emerging technique. Most of all, there is an urgent need to optimize the freeze/thaw procedure and to find the means to protect grafts from ischemia reperfusion injury. Nevertheless, ovarian transplantation should prove to be clinically useful for women at risk for premature ovarian failure. PMID- 11384625 TI - Spatial and temporal distribution of N-type Ca(2+) channels in gerbil global cerebral ischemia. AB - In the present study, we have investigated the spatial and temporal distribution of voltage-gated calcium channels in the gerbil model of global cerebral ischemia using immunohistochemistry. Distinct localizations of P-type (alpha(1A)), N-type (alpha(1B)), and L-type (alpha(1C) and alpha(1D)) Ca(2+) channels were observed in the hippocampus at days 1-5 after ischemic injury. However, increased expression of N-type Ca(2+) channels was detectable in brain regions vulnerable to ischemia only at days 2 and 3 after ischemic injury. The pyramidal cell bodies of CA1-3 areas and the granule cell bodies of the dentate gyrus were intensely stained at days 2 and 3 following ischemic injury. Transient changes in N-type Ca(2+) channel expression were also observed in the affected cerebral cortex and striatum at days 2 and 3 after ischemic injury. Although the present study has not addressed the multiple mechanisms contributing to the intracellular free Ca(2+) concentration ([Ca(2+)](i)) increase in the ischemic brain, the first demonstration of the transient increase in N-type Ca(2+) channels may prove useful for future investigations. PMID- 11384627 TI - Human reproductive cloning: thinking about clinic-based ethics. PMID- 11384628 TI - Lack of effect of isoflavonic phytoestrogen intake on leptin concentrations in premenopausal and postmenopausal women. AB - OBJECTIVE: To assess the effect of soy isoflavone ingestion on plasma leptin concentrations in premenopausal and postmenopausal women. DESIGN: Randomized, crossover studies, with blinding of participants and laboratory personnel. SETTING: Procedures involving free-living individuals were carried out at the University of Minnesota General Clinical Research Center. PATIENT(S): Fourteen regularly cycling premenopausal women, and 18 postmenopausal women. INTERVENTION(S): Each premenopausal participant consumed, on a daily basis, each of three soy protein powders containing different levels of isoflavones for three menstrual cycles plus 9 days, with plasma samples collected every other day the last 6 weeks of each diet period. Similarly, each postmenopausal participant consumed each of the three powders for 93 days, with plasma samples collected daily on days 64 to 66 and 92 to 94 of each diet period. The powders, dosed on a per-kilogram body weight basis, provided mean isoflavone intakes of 8, 65, and 130 mg/day, for the control, low-isoflavone, and high-isoflavone diet periods, respectively. MAIN OUTCOME MEASURE(S): Plasma leptin concentrations. RESULT(S): Isoflavone intake had essentially no effect on leptin concentrations in either premenopausal or postmenopausal participants. Concentrations in the premenopausal women were higher during the periovulatory and midluteal phases as compared to the early follicular and midfollicular phases. CONCLUSION(S): Despite the well documented effect of estrogens to enhance leptin production, even high levels of isoflavone consumption do not alter leptin concentrations in women. Further studies are needed to more precisely delineate the nature of estrogenic and/or antiestrogenic effects of isoflavones in humans. PMID- 11384629 TI - Relief of vasomotor symptoms and vaginal atrophy with lower doses of conjugated equine estrogens and medroxyprogesterone acetate. AB - OBJECTIVE: To evaluate the efficacy of lower doses of conjugated equine estrogens (CEE) plus medroxyprogesterone acetate (MPA) for relieving vasomotor symptoms and vaginal atrophy. DESIGN: A randomized, double-blind, placebo-controlled trial (the Women's Health, Osteoporosis, Progestin, Estrogen study). SETTING: Study centers across the United States. PATIENT(S): Two thousand, six hundred, seventy three healthy, postmenopausal women with an intact uterus, including an efficacy evaluable population (n = 241 at baseline). INTERVENTION(S): Patients received for 1 year (13 cycles; in milligrams per day) CEE, 0.625; CEE, 0.625 and MPA, 2.5; CEE, 0.45; CEE, 0.45 and MPA, 2.5; CEE, 0.45 and MPA, 1.5; CEE, 0.3; CEE, 0.3 and MPA, 1.5; or placebo. MAIN OUTCOME MEASURE(S): Number and severity of hot flushes and Papanicolaou smear with vaginal maturation index (VMI) to assess vaginal atrophy. RESULT(S): In the efficacy-evaluable population, reduction in vasomotor symptoms was similar with CEE of 0.625 mg/d and MPA of 2.5 mg/d (the most commonly prescribed doses) and all lower combination doses. CEE of 0.625 mg/d alleviated hot flushes more effectively than the lower doses of CEE alone. VMI improved in all active treatment groups. CONCLUSION(S): Lower doses of CEE plus MPA relieve vasomotor symptoms and vaginal atrophy as effectively as commonly prescribed doses. PMID- 11384630 TI - Effects of lower doses of conjugated equine estrogens and medroxyprogesterone acetate on endometrial bleeding. AB - OBJECTIVE: To evaluate vaginal bleeding profiles with lower doses of conjugated equine estrogens (CEE) and medroxyprogesterone acetate (MPA) as continuous combined therapy. DESIGN: The Women's Health, Osteoporosis, Progestin, Estrogen (Women's HOPE) study, a randomized, double-blind, placebo-controlled trial. SETTING: Study centers across the United States. PATIENT(S): Two thousand six hundred seventy-three healthy, postmenopausal women. INTERVENTION(S): Women received CEE, 0.625 mg/d; CEE, 0.625 mg/d, plus MPA 2.5 mg/d; CEE, 0.45 mg/d; CEE, 0.45 mg/d, plus MPA, 2.5 mg/d; CEE 0.45 mg/d, plus MPA, 1.5 mg/d; CEE, 0.3 mg/d; CEE, 0.3 mg/d, plus MPA, 1.5 mg/d; or placebo for 1 year. MAIN OUTCOME MEASURE(S): Bleeding data were analyzed in efficacy-evaluable and intention-to treat populations. RESULT(S): Cumulative amenorrhea and no bleeding rates were higher with lower doses of CEE/MPA than with CEE 0.625/MPA 2.5. A linear trend between time since menopause and cumulative amenorrhea was observed (P<.05) in all CEE/MPA groups except the CEE 0.45/MPA 1.5 group. The proportion of patients who experienced no bleeding in cycle 1 was 89%, 82%, and 80% in the CEE 0.3/MPA 1.5, CEE 0.45/MPA 1.5, and CEE 0.45/MPA 2.5 groups, respectively. These values were significantly greater than the incidence of no bleeding in the CEE 0.625/MPA 2.5 group (P<.05). CONCLUSION(S): Lower-dose regimens of CEE and MPA produce higher rates of amenorrhea and no bleeding compared with CEE 0.625/MPA 2.5 and may be appropriate for newly menopausal patients. PMID- 11384631 TI - Intratesticular Doppler flow, seminal plasma nitrites/nitrates, and nonobstructive sperm extraction from patients with obstructive and nonobstructive azoospermia. AB - OBJECTIVE: To prospectively evaluate the role of intratesticular vascular flow in modulating sperm function in men with obstructive and nonobstructive azoospermia. The correlation of testicular Doppler values with nitric oxide and testicular sperm extraction was further evaluated. DESIGN: Prospective study. SETTING: Assisted reproduction unit at a university center. PATIENT(S): Twenty-eight men with azoospermia undergoing sperm extraction for intracytoplasmic sperm injection. INTERVENTION(S): Ultrasound and color Doppler scanning of the testes. Testicular sperm retrieval and nitrite/nitrate assay. MAIN OUTCOME MEASURE(S): Doppler analysis of testicular transmediastinal artery, plasma and seminal plasma nitrite/nitrate values, and sperm extraction histopathology. RESULT(S): The pulsatility index (PI) of the transmediastinal artery was higher in patients with nonobstructive azoospermia (PI = 1.40 +/- 0.13) than in those with obstructive azoospermia (PI = 1.09 +/- 0.15; P=.011). Seminal plasma nitrite/nitrate concentrations were more elevated in cases of obstructive azoospermia than in gonadal failure. Unsuccessful sperm recovery was observed in four patients who showed the worst indices of gonadal failure. In this subgroup, a transmediastinal PI value >1.50 was always observed. CONCLUSION(S): Doppler analysis of the transmediastinal artery and nitrite/nitrate seminal plasma concentrations are useful for distinguishing between obstructive and nonobstructive azoospermia and allow the identification of the presence of spermatozoa within the testes. PMID- 11384632 TI - Intracytoplasmic sperm injection could minimize the incidence of prematurely condensed human sperm chromosomes. AB - OBJECTIVE: To compare the fertilization and prematurely condensed human sperm chromosomes (PCCs) rates between two intracytoplasmic sperm injection (ICSI) techniques. DESIGN: A retrospective study. SETTING: The data were obtained from the University of British Columbia in vitro fertilization (IVF) laboratory. PATIENT(S): ICSI cycles (n = 105) were performed for couples suffering from severe male-factor infertility and dysfunction of fertilization. INTERVENTION(S): Two types of ICSI techniques were used for ICSI procedures. MAIN OUTCOME MEASURE(S): Fertilization and pregnancy rates in group B using the improved ICSI technique were compared with those of group A using the standard ICSI technique. Unfertilized oocytes from the two groups were studied with cytogenetic methods. RESULT(S): Oocyte damage dropped from 14.8% in group A to 5.3% in group B. Normal fertilization for each group was 57.3% and 88.4%, respectively (P<.05). Pregnancy rate per egg retrieval was 15.6% in group A and 27.4% in group B (P<.05). PCCs occurred in 19.4% of unfertilized oocytes in group A and did not occur in group B. CONCLUSION(S): This study indicates that ICSI not only yields high fertilization rates, but also minimizes the incidence of PCCs. It may be directly related to two crucial steps (immobilization of sperm and aspiration of oocyte cytoplasm) used in ICSI procedures. This study also suggests that it is possible to overcome one cause of IVF failure resulting from the formation of PCCs by using the improved ICSI technique. PMID- 11384633 TI - Testicular ultrasonography and extended chromosome analysis in men with nonmosaic Klinefelter syndrome: a prospective study of possible predictive factors for successful sperm recovery. AB - OBJECTIVE: To investigate whether extended chromosome analysis or testicular sonography, including flow Doppler imaging, before diagnostic testicular sperm extraction have predictive value for successful sperm retrieval in men with nonmosaic Klinefelter syndrome. DESIGN: Prospective clinical study. SETTING: IVF clinic and genetics laboratory at a university hospital. PATIENT(S): Nineteen patients with nonmosaic Klinefelter syndrome and azoospermia. INTERVENTION(S): Collection of blood samples; histopathologic examination of testicular tissue; fluorescence in situ hybridization; sonography, including Doppler imaging; and testicular sperm extraction. MAIN OUTCOME MEASURE(S): Testicular volume, serum FSH and serum testosterone levels, percentage of normal XY cells, ultrasound echogenicity, intratesticular blood flow resistance, and sperm recovery. RESULT(S): Testicular volume and levels of serum FSH and serum testosterone levels did not differ significantly. No differences in testicular echogenicity or intratesticular blood flow resistance were found between 47,XXY men in whom sperm recovery was successful and those in whom sperm recovery failed. Significant differences were seen between all patients with the Klinefelter syndrome and controls with normal sperm values. Fluorescence in situ hybridization of peripheral lymphocytes and buccal tissue showed no correlation between frequency of normal 46,XY cells and testicular spermatogenesis. CONCLUSION(S): In azoospermic men with the Klinefelter syndrome, histopathologic findings seem to be predictive for successful sperm recovery. Infertility work-up, including diagnostic testicular sperm recovery, is recommended, and, if possible, viable sperm should be cryopreserved. PMID- 11384634 TI - Recombinant or urinary follicle-stimulating hormone? A cost-effectiveness analysis derived by particularizing the number needed to treat from a published meta-analysis. AB - OBJECTIVE: To demonstrate that particularizing pooled results of a meta-analysis can derive incremental cost effectiveness of superovulation with recombinant follicle-stimulating hormones (rFSH) vs. the highly purified urinary form (uFSH) for assisted conception. DESIGN: A retrospective study. SETTING: An assisted conception unit in the United Kingdom. PATIENT(S): One hundred forty-five fresh in vitro fertilization (IVF) and 58 fresh intracytoplasmic sperm injection (ICSI) cycles. INTERVENTION(S): rFSH vs. uFSH. MAIN OUTCOME MEASURE(S): Incremental cost effectiveness (i.e., cost needed to treat, or CNT) and budget-impact analyses of rFSH vs. uFSH. RESULT(S): In women less than 30 years old, the clinical pregnancy rate was 37.7% (95% CI 24.8%-52.1%), the particularized number needed to treat (pNNT) was -19, and the cost needed to treat was 5070.51 pounds sterling (3660.53 pounds sterling to 7619.32 pounds sterling). For the 30- to 35-year-old age group, the clinical pregnancy rate was 29.9% (95% CI 20.0%--41.4%), the particularized number needed to treat was -24, and CNT was 7335.59 pounds stering (5284.11 pounds sterling to 10,941.22 pounds sterling). For the 36- to 40-year old age group, the clinical pregnancy rate was 30.6.0% (95% CI 19.6%--43.7%), the particularized number needed to treat was -23.0, and the CNT was 8569.67 pounds sterling (5998.70 pounds sterling to 13,413.24 pounds sterling). CONCLUSION(S): The CNT and thus the budget impact analyses (the extra number of cycles that can be funded by the CNT) both increase directly with age of the patient, and inversely with the clinical pregnancy rate. PMID- 11384635 TI - Induction of ovulation in World Health Organization group II anovulatory women undergoing follicular stimulation with recombinant human follicle-stimulating hormone: a comparison of recombinant human chorionic gonadotropin (rhCG) and urinary hCG. AB - OBJECTIVE: To compare the safety and efficacy of 250 microg recombinant hCG (rhCG) and 5,000 IU urinary hCG (uhCG), both administered s.c., for ovulation induction in anovulatory/oligo-ovulatory patients after follicular stimulation with recombinant hFSH (rhFSH). DESIGN: Phase III, double-blind, double-dummy, randomized, parallel-group, multicenter study. SETTING: Nineteen academic and private tertiary care infertility centers in Europe, Israel, Canada, and Australia. PATIENT(S): One hundred ninety-eight WHO group II anovulatory women, aged 20 to 38 years. INTERVENTION(S): Women were randomized to receive rhCG or uhCG after follicular stimulation with rhFSH in a chronic low-dose protocol. Blood samples were collected and ultrasound examinations performed during stimulation and after hCG administration. MAIN OUTCOME MEASURE(S): Ovulation (midluteal serum progesterone > or =30 nmol/L), serum progesterone, hCG levels after hCG, pregnancy, adverse events, local tolerability, and ovarian hyperstimulation syndrome (OHSS) incidence. RESULT(S): Ovulation rates did not differ between groups: 95.3% for rhCG (n = 85) and 88.0% for uhCG (n = 92). The one-sided 95% confidence interval for the observed difference fell above the predefined limit of -20%, indicating equivalence. Treatment was well tolerated, but more uhCG patients reported local reactions (particularly inflammation and pain) (P=.0001; logistic regression). CONCLUSION(S): Subcutaneous rhCG and uhCG show equivalent efficacy in ovulation induction; however, rhCG is better tolerated. PMID- 11384636 TI - Psychopathology, personality, and marital relationship in patients undergoing in vitro fertilization procedures. AB - OBJECTIVE: To compare the psychopathology, personality features, and marital relationships of women undergoing in vitro fertilization (IVF) with those of control patients, and to compare IVF inductees with program veterans. DESIGN: Cross-sectional clinical study. SETTING: A university hospital. PATIENT(S): One hundred and one women undergoing IVF treatment. INTERVENTION(S): Psychometric tests were administered at first visit (baseline) of index treatment cycle. MAIN OUTCOME MEASURE(S): Achievement of pregnancy. RESULT(S): Women undergoing IVF show higher levels of anxiety and emotional tension than do controls. Although the infertile women showed no abnormal personality dimensions, the IVF group did have a particular psychological profile and a different marital relationship pattern when compared with the control participants. Between IVF veterans and inductees, there are significant differences with respect to psychopathology, psychological dimensions, and couple dynamics. The achievement of pregnancy is not associated with any special psychopathological, personality, or marital characteristics among the IVF women. CONCLUSION(S): The most crucial period in IVF procedures may immediately follow the end of the first cycle because of the high risk of patients dropping out of the program. To determine the most effective supporting therapies for women undergoing fertilization procedures it could be useful to consider the psychological and relational differences between veterans and inductees. PMID- 11384637 TI - A comparison of day 5 and day 6 blastocyst transfers. AB - OBJECTIVE: To compare implantation and pregnancy rates according to the day of embryo transfer (day 5 or 6 after oocyte retrieval) when transfer was postponed until expanded blastocysts developed. DESIGN: Retrospective clinical study. SETTING: Private ART center. PATIENT(S): One-hundred and eighty-three women undergoing blastocyst-stage embryo transfer following in vitro fertilization. INTERVENTION(S): Bipronucleate oocytes were grown for up to 144 hours and subsequently transferred only when at least one embryo attained the expanded blastocyst stage. MAIN OUTCOME MEASURE(S): Implantation and pregnancy rates. RESULT(S): Blastocysts transferred on day 5 implanted at nearly twice the rate of blastocysts transferred on day 6 (36.3% vs. 19.0%). Pregnancy rates were also almost twice as high among the day 5 transfer patients (59.3% vs. 32.3%). In addition, more blastocysts developed (3.6 vs. 2.4), and more were transferred (2.7 vs. 2.3) to the day 5 transfer patients, although the proportion of expanded blastocysts among the blastocysts that were transferred was the same for the two groups (91.7% vs. 93.6%). CONCLUSION(S): Embryos that develop to the expanded blastocyst stage and are transferred on day 5 after retrieval are approximately twice as likely to implant compared to those for which expansion and transfer are delayed until day 6. PMID- 11384638 TI - A low-dose stimulation protocol using highly purified follicle-stimulating hormone can lead to high pregnancy rates in in vitro fertilization patients with polycystic ovaries who are at risk of a high ovarian response to gonadotropins. AB - OBJECTIVE: To study the benefits of a low-dose stimulation (LDS) protocol with purified urinary follicle-stimulating hormone in patients with polycystic ovaries who have presented previously with a very high ovarian response to a standard hMG stimulation. DESIGN: Cohort study. SETTING: Fertility center in a university hospital. PATIENT(S): Sixty-one patients involved in an IVF/ICSI program from January 1995 to December 1996. INTERVENTION(S): The patients were first stimulated with a standard protocol using hMG and presented with a very high ovarian response. These patients were then stimulated a second time using a low dose protocol. Cryopreserved embryos were transferred in later artificial or natural cycles until to December 1999. MAIN OUTCOME MEASURE(S): Number of gonadotropin ampules; estradiol level on the day of ovulation induction; follicles, oocytes, and cryopreserved zygotes; fertilization, implantation, and pregnancy rates; and number of ovarian hyperstimulation syndromes (OHSS). RESULT(S): The number of ampules used, the estradiol level reached, and the number of oocytes obtained were significantly lower under the LDS than the standard protocol. High implantation (21.8%) and clinical pregnancy (38.4%) rates were obtained after LDS. The cumulated deliveries per cycle started and per patient were, respectively, 41.6% and 52.5%. Five patients suffered OHSS with the standard protocol, and none with the LDS. CONCLUSION(S): The LDS protocol offers a safe and efficient treatment for patients who present with echographic polycystic ovaries and are at risk of an excessive ovarian response to standard IVF stimulation protocols. PMID- 11384639 TI - Effects of vaginal progesterone administration on uterine contractility at the time of embryo transfer. AB - OBJECTIVE: To investigate whether uterine contractility at the time of embryo transfer (ET) can be reduced by early onset of luteal support with progesterone administered vaginally. DESIGN: Prospective analysis. SETTING: Assisted reproduction unit. PATIENT(S): Eighty-four women undergoing 84 GnRH-a and FSH/hCG cycles for IVF-ET were studied. INTERVENTION(S): Vaginal progesterone was randomly started on the day of oocyte retrieval (group A, n = 43) or on the evening of ET (group B, n = 41). On the day of hCG administration and just before ET, 2-minute sagittal uterine scans were obtained by ultrasound and digitized with an image analysis system for assessing uterine contraction frequency. MAIN OUTCOME MEASURE(S): Uterine contraction frequency. RESULT(S): Whereas uterine contraction frequency was similar in both groups on the day of hCG (4.6 +/- 0.3 and 4.5 +/- 0.3 contractions per minute, respectively), only women in group A showed decreased uterine contraction frequency on the day of ET (2.8 +/- 0.2 vs. 4.2 +/- 0.3 contractions per minute). CONCLUSION(S): Vaginal progesterone administration starting on the day of oocyte retrieval induced a decrease in uterine contraction frequency on the day of ET as compared with preovulatory values. Uterine relaxation before ET is likely to improve IVF-ET outcome by avoiding the displacement of embryos from the uterine cavity. PMID- 11384640 TI - Increase in nerve fibers and loss of mast cells in polycystic and postmenopausal ovaries. AB - OBJECTIVE: To quantify nerve fibers and mast cells in human ovaries at different functional stages. DESIGN: Retrospective study. SETTING: Research laboratory of the university. SPECIMEN(S): 8 human ovaries in the follicular (cyclic) phase, 7 polycystic ovaries, and postmenopausal ovaries with (n=5) or without (n=7) hyperthecosis. MAIN OUTCOME MEASURE(S): Single- and double immunohistology for the S100 antigen in glial cells of autonomic nerve fibers, for chymase and tryptase in mast cells, and for the common leukocyte antigen on leukocytes. Histometric evaluation was also performed. INTERVENTION(S): None. RESULT(S): Polycystic ovaries contained significantly more S100-positive nerve fibers in the corticomedullary region than did cyclic ovaries (mean +/- SD per 2-mm(2) area, 476 +/- 136 and 224 +/- 133; P<.01). Postmenopausal ovaries with or without hyperthecosis had the highest density of nerve fibers. In cyclic and polycystic ovaries, more tryptase-positive mast cells than chymase-positive mast cells were found in the interstitial cortex and the medulla. In cyclic ovaries, areas with a moderate density of nerve fibers contained many mast cells. Hence, with increasing nerve fiber density in polycystic ovaries, the number of mast cells decreased strikingly compared with cyclic ovaries (p<.001). Almost no mast cells were seen in postmenopausal ovaries with and without hyperthecosis. The number of leukocyte antigen-positive leukocytes was similar in all groups. CONCLUSION(S): The high density of nerve fibers in polycystic and postmenopausal ovaries, together with a conspicuous decrease in mast cells, indicates altered neuroimmune communication. PMID- 11384641 TI - Familial gonadotropin-releasing hormone resistance and hypogonadotropic hypogonadism in a family with multiple affected individuals. AB - OBJECTIVE: To characterize the phenotype of idiopathic hypogonadotropic hypogonadism due to compound heterozygous GnRHR gene mutations (Arg262Gln/Tyr284Cys). DESIGN: Retrospective review. SETTING: Tertiary medical center. PATIENT(S): Family containing four siblings (three female and one male) with complete idiopathic hypogonadotropic hypogonadism. INTERVENTION(S): Baseline and stimulated laboratory studies. One patient received GnRH treatment and one received human menopausal gonadotropins. MAIN OUTCOME MEASURE(S): Clinical phenotype vs. genotype is assessed by endocrine studies, karyotype, pedigree, and review of pathology slides of ovarian neoplasm. RESULT(S): With GnRH stimulation, two patients with idiopathic hypogonadotropic hypogonadism had maximum LH < 10 mIU/mL, and two others had peak LH > 10 mIU/mL. With repeated GnRH stimulation 24 hours later, gonadotropin levels in all patients were increased. Stimulation of thyroid-releasing hormone and tests for insulin-induced hypoglycemia were normal. One affected patient did not ovulate after GnRH treatment, but her sister ovulated with gonadotropin treatment. Another affected sibling had bilateral oophorectomy for seromucinous cystadenomas, and her hypogonadotropic state remained after castration. The man with idiopathic hypogonadotropic hypogonadism and his unaffected brother had a ring chromosome 21. CONCLUSION(S): All patients with complete idiopathic hypogonadotropic hypogonadism had the same GnRHR mutations, but clinical presentations and endocrinologic responses were heterogeneous. Gonadotropin levels remained low in patients with idiopathic hypogonadotropic hypogonadism after castration, and ring chromosome 21 was present, suggesting that sequences from this chromosome could affect the idiopathic hypogonadotropic hypogonadism phenotype. PMID- 11384642 TI - Is there evidence for preferential delivery of ovarian estradiol to the endometrium? AB - OBJECTIVE: To determine the direction of delivery of E(2) in the female pelvis by assessing the ratio of endometrial to serum E(2) in women whose ovaries were stimulated to produce E(2) with women who received exogenous E(2). DESIGN: Prospective comparative study. SETTING: University-based ART program. PATIENT(S): Oocyte donors and recipients of donor oocytes. INTERVENTION(S): Micronized E(2) administered by the oral or vaginal route and oocyte donation. MAIN OUTCOME MEASURE(S): Serum and endometrial levels of E(2). RESULT(S): Serum E(2) levels were significantly higher in women who underwent controlled ovarian hyperstimulation (COH) and women receiving exogenous E(2) by the vaginal route than in those who received oral E(2). Levels of E(2) in endometrial tissue were similar in women who underwent COH and those receiving oral E(2). Endometrial E(2) levels in women who underwent vaginal administration were significantly higher than those in the oral E(2) or COH groups. The ratio of endometrial to serum E(2) was highest in women who underwent vaginal E(2) and lowest in those undergoing COH. CONCLUSION(S): Vaginal administration of micronized E(2) results in preferential absorption of E(2) into the endometrium, consistent with a "uterine first pass" effect. Since endogenous E(2) produced the smallest ratio of E(2) between the endometrium and serum, E(2) produced by the ovaries is not preferentially delivered to the uterus. PMID- 11384643 TI - Effects of dioxin, an environmental pollutant, on mouse blastocyst development and apoptosis. AB - OBJECTIVE: To evaluate the effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; dioxin) on mouse embryo development and apoptosis. DESIGN: Controlled animal study. SETTING: Academic research environment. ANIMAL(S): Female mice (CB6F1) at 3 to 6 weeks of age and proven breeders (C578B46). INTERVENTION(S): Mouse embryos were obtained at the morula stage and cultured to the blastocyst stage in a pharmacologic dose of TCDD (3.1 microM) or a control medium. The morphology was assessed, and staining for apoptosis was performed. Immunohistochemistry for the presence of aromatic hydrocarbon receptor (AhR) was performed in another set of morula-stage embryos. MAIN OUTCOME MEASURE(S): The number of embryos developing from the morula to the blastocyst stage and number of apoptotic blastomeres in control vs. TCDD culture conditions. RESULT(S): No statistically significant differences were observed in the percentage of embryos reaching the blastocyst stage: 80.9% (115 of 142) in the TCDD-treated group, vs. 82.9% (121 of 146) in the control group. There was also no difference in the degree of apoptosis: 22.6 +/- 7.3% apoptotic cells (TCDD) vs. 25.3 +/- 9.7% (controls). Staining indicated the slight presence of aromatic hydrocarbon receptor in the morula-stage mouse embryos. CONCLUSION(S): TCDD at 3.1 microM did not alter the development of early mouse morula to blastocysts and did not significantly induce apoptosis in vitro. PMID- 11384644 TI - Nitric oxide as a regulator in preimplantation embryo development and apoptosis. AB - OBJECTIVE: To investigate the mechanisms of nitric oxide (NO) in the development and apoptosis of preimplantation mouse embryos. DESIGN: Prospective, controlled study. SETTING: Medical college laboratory. SUBJECT(S): Two-cell embryos from outbred ICR mice. INTERVENTION(S): Hyperstimulation protocol, two-cell embryos were collected, then treated with or without an NO synthase inhibitor (L-NAME) or an NO donor (SNP) and combined with a cGMP analogue (8-Br-cGMP) or a selective inhibitor of NO-sensitive soluble guanylyl cyclase (ODQ). MAIN OUTCOME MEASURE(S): The development of ICR mouse embryo from two cells to blastocyst stages in vitro. RESULT(S): The development of blastocyst was inhibited by L-NAME in a concentration-dependent manner (0.1-10 microM) and 0.1 microM SNP reversed this effect (80.5% of control). Annexin-V/propidium iodide and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling techniques demonstrated that excessive NO (> or =10 microM) might induce apoptosis in the mouse embryos. 8-Br-cGMP reversed the inhibitory effect of L-NAME and rescued the embryo growth. ODQ inhibited the embryo development in a dose-responsive fashion (0.1--100 microM) but had no effect in the NO-induced embryo apoptosis. P53 and Bax were found to be up-regulated during the embryo fragmentation. CONCLUSION(S): These results indicate that the cGMP pathway might be involved in the NO regulated embryonic development, but not in NO-induced apoptosis, for which P53/Bax pathway might be involved. PMID- 11384645 TI - Expression of aminopeptidase N in human endometrium and regulation of its activity by estrogen. AB - OBJECTIVES: To determine whether aminopeptidase N (APN) regulates the cycle dependent bioavailability of interleukin-8 (IL-8) in the endometrium. DESIGN: Prospective study. SETTING: University medical center. PATIENT(S): Women without endometrial pathology from the proliferative (n = 25) or secretory (n = 18) phase of the menstrual cycle. INTERVENTION(S): We first immunolocalized APN in the endometrium using an anti-APN antibody. We then determined the regulation of APN kinetic activity by sex steroids in endometrial stromal cell cultures. MAIN OUTCOME MEASURE(S): Expression of APN in human endometrium throughout the menstrual cycle. Regulation of APN activity by estradiol and progesterone in cultured endometrial stromal cells. RESULT(S): Immunohistochemistry of endometrial sections revealed staining of endometrial stroma throughout the menstrual cycle. There was no detectable staining in glandular cells. The expression of APN as detected by immunohistochemistry was significantly lower in the early proliferative phase. In cultured cells, estradiol inhibited APN activity in a concentration-dependent manner. Progesterone did not have a significant effect. CONCLUSION(S): Stromal localization of APN in endometrium may explain the epithelial rather than stromal presence of IL-8 in vivo. Decreased expression of APN may increase IL-8 bioavailability thus contributing to angiogenesis and polymorphonuclear leukocyte chemotaxis in early proliferative phase. PMID- 11384646 TI - Cryopreservation of ICR mouse oocytes: improved post-thawed preimplantation development after vitrification using Taxol, a cytoskeleton stabilizer. AB - OBJECTIVE: To establish an effective cryopreservation method. DESIGN: In vitro model study. SETTING: Infertility Medical Center, Pochon CHA University. ANIMAL(S): Four-week-old ICR mice superovulated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin. INTERVENTION(S): Vitrified thawed oocytes were fertilized and subsequently cultured in vitro. MAIN OUTCOME MEASURE(S): Post-thawed development, chromosome/spindle normalities, and blastocyst quality. RESULT(S): More cumulus-enclosed oocytes were fertilized and developed to the 8-cell stage after vitrification and thawing than denuded oocytes. However, cryopreserved oocytes of both types had lower spindle and chromosome normalities than fresh oocytes, which resulted in reduced developmental competence after thawing. The addition of 1 microM of Taxol, a cytoskeleton stabilizer, to vitrification solution greatly promoted the blastocyst formation of vitrified-thawed oocytes, compared with no addition (24.0% vs. 58.6%). No difference in blastocyst quality, which was evaluated by blastomere and inner cell mass cell numbers and inner cell mass cell per trophoblast ratio, was found between fresh oocytes and oocytes vitrified with Taxol. CONCLUSION(S): A vitrification solution consisting of 5.5 M ethylene glycol, 1.0 M sucrose, 10% fetal bovine serum, and 1 microM Taxol greatly improved post-thawed development of vitrified oocytes. PMID- 11384647 TI - A novel system for in vitro maturation of human oocytes. AB - OBJECTIVE: To compare in vitro maturation of cumulus-free oocytes in glucose-free medium (P1) and standard medium (TC199). DESIGN: Prospective, cohort study. SETTING: Assisted reproductive technology program. PATIENT(S): One hundred eight patients undergoing ICSI. INTERVENTION(S): Germinal vesicle-stage or metaphase I- stage oocytes were allocated to culture with P1 or TC199. Metaphase II oocytes were fixed for immunofluorescence analysis or fluorescence in situ hybridization at 24 or 48 hours (or both). Media were compared by performing conditional logistic regression analysis that controlled for egg-specific factors. MAIN OUTCOME MEASURE(S): Proportion of mature oocytes and appearance of normal spindle chromosome cytoarchitecture. RESULT(S): At 24 hours, more P1 oocytes than TC199 oocytes reached metaphase II (59.7% vs. 44.9%). At 48 hours, 71.7% of P1 oocytes and 61.0% of TC199 oocytes reached metaphase II, but this difference was not significant. Metaphase II oocytes in P1 were 34.3% more likely than those in TC199 to have a bipolar spindle with aligned chromosomes. Compared with oocytes at the germinal vesicle stage at 0 hour, those at metaphase I at 0 hour were more likely to progress to metaphase II (72.6% vs. 46.1% at 24 hours; 84.1% vs. 60.6% at 48 hours). CONCLUSION(S): P1 is superior to TC199 for in vitro maturation of granulosa-free human oocytes. PMID- 11384648 TI - Rescue of mouse embryos from 2-cell blocks by microinjection of maturation promoting factor. AB - OBJECTIVE: To examine the rescue of mouse embryos from 2-cell blocks by the microinjection of maturation promoting factor (MPF) extracted from matured Xenopus eggs into one of the blastomeres of 2-cell stage mouse embryos. DESIGN: Controlled laboratory study. SETTING: First Department of Obstetrics and Gynecology, Toho University School of Medicine, Tokyo, Japan. ANIMAL(S): Eight- to 10-week-old female Crj:CD-1(ICR) mice. INTERVENTION(S): One of the blastomeres of the mouse 2-cell embryos was injected with MPF (MI group) or mHTF medium (MED group) at 28--32 hours after insemination. MAIN OUTCOME MEASURE(S): The developmental rate to blastocyst. RESULT(S): The developmental rate to blastocyst in the MI group (48.0%) was significantly higher than that in the MED group (0%). CONCLUSION(S): The 2-cell block was specifically rescued by the microinjection of MPF and not by the insertion of pipettes. PMID- 11384649 TI - Expression of estrogen receptor alpha and beta in peritoneal and ovarian endometriosis. AB - OBJECTIVE: To quantify and compare messenger RNA (mRNA) levels of ER-alpha and ER beta among ovarian endometriotic cysts and red and black peritoneal endometriotic lesions. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Patients with or without endometriosis. INTERVENTION(S): Samples of peritoneal (n = 33) and ovarian endometriotic lesions (n = 37) were obtained during laparoscopic surgery. Normal eutopic endometrial tissues and macroscopically normal peritoneal tissues were obtained as controls during or just after surgery. MAIN OUTCOME MEASURE(S): Expression of mRNA for ER-alpha and ER-beta, using a real-time reverse transcription (RT)-PCR assay, TaqMan RT-PCR, and nonradioactive in situ hybridization (ISH) techniques. RESULT(S): Both eutopic endometrium and endometriotic tissues showed predominantly higher levels of ER-alpha than ER-beta mRNA. Relative ratio of ER-alpha to ER-beta (ER-alpha/ER-beta) mRNA in red peritoneal lesions was significantly higher than in black lesions and ovarian endometriotic cysts. There was no significant difference in ER-alpha/ER-beta between proliferative eutopic endometrium and red peritoneal lesions. These results were confirmed by ISH analysis, which also revealed that the two estrogen receptors were localized in both epithelial and stromal cells of endometriotic tissues. CONCLUSION(S): The predominant expression of ER-alpha in both glandular epithelial and stromal cells may be essential for the development and growth of peritoneal and ovarian endometriosis. PMID- 11384650 TI - Influence of the angiotensinogen gene on the ovulatory capacity of mice. AB - OBJECTIVE: The tissue-bound ovarian renin-angiotensin system (OVRAS) is critically involved in ovulation in humans and rodents. Mice with disruption and overexpression of the angiotensinogen gene (Agt) have been previously generated. We investigated the influence of varying Agt gene expression on the ovulatory capacity and early embryonic development in mice. DESIGN: Observational study of genetically altered mice and their response to a superovulation protocol. SETTING: Academic research institution. ANIMAL(S): Mice with varying copy numbers of Agt (one copy: n = 48; two copies: n = 51; three copies: n = 20; four copies: n = 24). INTERVENTION(S): Superovulation protocol, oocyte culture. MAIN OUTCOME MEASURE(S): Number of oocytes harvested, early embryonic development of zygotes, evaluation of ovarian histology, serum estradiol measurements. RESULT(S): The mean number of oocytes harvested was greatest in wild-type mice (two copies of Agt, 39.9 +/- 14) with a reduction of ovulatory capacity in mice overexpressing Agt (three copies [34.8 +/- 11.7] and four copies [31.2 +/- 12.4], P =.026). Mice with one copy of Agt showed a slight decrease of ovulatory capacity compared to wild-type mice (35.8 +/- 15.2, P =.29). Ovarian histology, serum estradiol levels, and early embryonic development were independent of the Agt genotype. CONCLUSION(S): Overexpression of Agt was associated with reduced ovulatory capacity, but with none of the other parameters that were evaluated. These findings support an important role of the ovarian renin-angiotensin system in the process of follicular rupture. PMID- 11384651 TI - A technique for laparoscopic transplantation of frozen-banked ovarian tissue. AB - OBJECTIVE: To describe a laparoscopic technique for transplanting frozen-banked ovarian tissue. DESIGN: Case study. SETTING: Academic medical center. PATIENT(S): A patient whose ovarian tissue was previously frozen with the slow-freeze technique. INTERVENTION(S): Microsurgical reconstruction of ovarian cortex and its laparoscopic transplantation to the ovarian fossa. MAIN OUTCOME MEASURE(S): Blood flow to the grafts by doppler, follicle development and ovulation by ultrasound, E(2) and progesterone production, and resumption of spontaneous menses. RESULT(S): The patient ovulated and menstruated 4 months after the transplant in response to ovarian stimulation with menopausal gonadotropins. CONCLUSION(S): Laparoscopic transplantation of frozen-banked ovarian tissue beneath the pelvic peritoneum can restore ovarian function. PMID- 11384652 TI - Implantation of a gestational sac in a cesarean section scar. PMID- 11384653 TI - Vascular endothelial growth factor and interleukin-8 in ovarian cystic pathology. AB - OBJECTIVE: To determine the levels of the angiogenic factors vascular endothelial growth factor (VEGF) and interleukin (IL-8) in ovarian cysts. DESIGN: Prospective descriptive study. SETTING: University hospital. PATIENT(S): One hundred women, of whom 9 had ovarian carcinomas, 38 had ovarian endometriomata, 43 had serous ovarian cysts, and 10 had follicular ovarian cysts. INTERVENTION(S): Sampling of serum and ovarian cystic fluid before and during surgery. MAIN OUTCOME MEASURE: Levels of VEGF and IL-8 in cystic fluid and serum. RESULT(S): Levels of both VEGF and IL-8 were found to be significantly higher in the cystic fluid of ovarian carcinomas and endometriomata than in serous and follicular cysts. In endometriomata fluid, levels of VEGF and IL-8 were found to be directly correlated (r = 0.68; P=.0074). Serum levels of VEGF were significantly higher in women with ovarian carcinomas and endometriomata than in those with serous and follicular cysts. Ovarian cancers and endometriomata were similar in terms of cystic concentrations of VEGF and IL-8 and in serum levels of VEGF. CONCLUSION(S): An increase in angiogenic factors that differentiate ovarian carcinomas and endometriomata from other kinds of ovarian pathology is demonstrated. PMID- 11384654 TI - Chromosomal translocation 3;22 in an infertile man. AB - OBJECTIVE: To present the first case of an infertile male with a normal phenotype and chromosomal translocation 3;22. DESIGN: Case report. SETTING: POVISA Medical Center. PATIENT(S): A 45-year-old man with primary infertility for 13 years and with different partners; the patient has a family history of recurrent miscarriages and low fertility. INTERVENTION(S): Lymphocytic karyotype and electron microscopy. MAIN OUTCOME MEASURE(S): Physical examination and semen analysis. RESULT(S): The semen analysis revealed oligoasthenoteratospermia. The lymphocytic karyotype detected a translocation 3;22, and electron microscopy showed a lack of the central microtubule pair and peripheral doublet. CONCLUSION(S): An association between translocation 3;22 and other abnormalities in infertile males has been reported, but no such association has ever been described in men whose only clinical manifestation is infertility. PMID- 11384655 TI - A case of 46,X,der(X)(pter-->q21::p21-->pter) with gonadal dysgenesis, tall stature, and endometriosis. AB - OBJECTIVE: To report a case of 46,X,der(X)(pter-->q21::p21-->pter) with gonadal dysgenesis, tall stature, and endometriosis. DESIGN: Case report. SETTING: A university hospital. PATIENT(S): A 20-year-old primary amenorrheal woman receiving estrogen-progestogen substitution. INTERVENTION(S): G-banding, comparative genomic hybridization, fluorescence in situ hybridization (FISH), and laparoscopy. MAIN OUTCOME MEASURE(S): A recombinant X chromosome, 46,X,der(X)(pter-->q21::p21-->pter), and pelvic endometriosis. RESULT(S): The patient's chromosomal abnormality was misjudged by the use of G-banding as a distal part deletion of the long arm in one X chromosome. Comparative genomic hybridization and fluorescence in situ hybridization analyses with locus-specific probes revealed 46,X,der(X)(pter-->q21::p21-->pter). The laparoscopic examination showed bilateral streak gonads and blue berry spots at the pelvic peritoneum, which were confirmed by evaluation of biopsy specimens. CONCLUSION(S): Recent advances of genetic strategies make it easy to determine karyotype and phenotype abnormalities. We have to keep our mind on the potential of endometriosis with patients who are receiving estrogen-progestogen substitution. PMID- 11384656 TI - Should all infertile males undergo urologic evaluation before assisted reproductive technologies? Two cases of testicular cancer presenting with infertility. AB - OBJECTIVE: To report two cases of testicular cancer in patients presenting with infertility. DESIGN: Case reports. SETTING: University-affiliated urology practice. PATIENT(S): Two men presenting with infertility. INTERVENTION(S): Complete history and physical, hormonal assays, semen analysis, scrotal ultrasound, radical orchiectomy. MAIN OUTCOME MEASURE(S): Testicular pathology specimens. RESULT(S): Testicular cancer was diagnosed in two men sent to a urology clinic for infertility treatment. CONCLUSION(S): A thorough evaluation should be completed in all males in couples presenting with infertility. PMID- 11384657 TI - Hydroxyethylstarch versus human albumin for the treatment of severe ovarian hyperstimulation syndrome: a preliminary report. AB - OBJECTIVE: To compare the efficacy and safety of 6% hydroxyethylstarch and human albumin as colloid solutions for treatment of severe ovarian hyperstimulation syndrome (OHSS). DESIGN: Controlled cohort study. SETTING: Tertiary medical center. PATIENT(S): Sixteen patients with severe OHSS. INTERVENTION(S): Six percent hydroxyethylstarch (6 patients) and human albumin (10 patients). MAIN OUTCOME MEASURE(S): Urine output, number of abdominal and pleural drainage procedures, complications, duration of hospitalization, and perinatal outcome. RESULT(S): Patients who received 6% hydroxyethylstarch had higher urine output, needed fewer abdominal and pleural paracenteses, and had a shorter hospital stay than those who received human albumin. In each group, no adverse effects or congenital malformations were observed and the rates of miscarriage were similar. CONCLUSION(S): These results suggest that 6% hydroxyethylstarch may be superior to albumin as a colloid solution for the treatment of severe OHSS. PMID- 11384658 TI - Endometrial receptivity in terms of pinopode expression is not impaired in women with endometriosis in artificially prepared cycles. AB - OBJECTIVE: To assess endometrial receptivity in terms of pinopode expression in women with endometriosis. DESIGN: Prospective, observational study. SETTING: Oocyte donation program at the Instituto Valenciano de Infertilidad. PATIENT(S): Twelve women with endometriosis as the only cause of infertility. INTERVENTION(S): Pinopode expression pattern analysis by scanning electron microscopy in two sequential endometrial biopsies obtained in the same cycle of each patient. MAIN OUTCOME MEASURE(S): Pinopode pattern and pregnancy rates. RESULT(S): Pinopode expression in women with endometriosis did not differ from that of patients without endometriosis undergoing artificial cycles. Similarly, the clinical outcome in these women was comparable to that of the general population included in the oocyte donation program. The pregnancy rate per transfer was 46.7%. CONCLUSION(S): These results show that in women with endometriosis undergoing oocyte donation under hormone replacement therapy, pinopode expression is not altered, suggesting that endometrial receptivity in women with this disease remains unaltered. PMID- 11384659 TI - Progesterone increases the number of Langerhans cells in human vaginal epithelium. PMID- 11384660 TI - Ampullary, seminal vesicular, and prostatic massage for obtaining spermatozoa from patients with anejaculation. PMID- 11384661 TI - The luteinizing hormone beta-subunit exon 3 (Gly102Ser) gene mutation is rare in Korean women with endometriosis and polycystic ovary syndrome. PMID- 11384662 TI - Invasive prenatal diagnosis after intracytoplasmic sperm injection. PMID- 11384663 TI - Effects of serial thaw-refreeze cycles on human sperm motility and viability. PMID- 11384664 TI - Specific recognition of cytosolic thymidine kinase in the human lung tumor by monoclonal antibodies raised against recombinant human thymidine kinase. AB - Anti-TK monoclonal antibodies (mAbs) were raised against recombinant human cytosolic thymidine kinase (rhTK) and characterized by Western immunoblotting, enzyme-linked immunosorbent assay (ELISA) and immunostaining of tumor cells. Twenty-three clones of TK mAbs were characterized to recognize specifically not only rhTK produced by Escherichia coli but also TK subunit of 25 kDa in human lung cancer. The anti-TK mAbs reacted specifically with cytosolic TK but not with mitochondrial TK. Only one clone of the mAbs inhibited the catalytic activity of TK. By solid phase sandwich enzyme immunoassay using these mAbs, we could quantitate the cytosolic TK content in tissues. Immunohistochemical staining analysis using one of the TK mAbs showed that human lung adenocarcinoma and squamous cell carcinoma exhibited much higher staining intensity than stromal cells. These mAbs are useful for biochemical studies on the regulation of human TK in proliferating cells such as tumor cells and for diagnosis of highly proliferating tumors. PMID- 11384665 TI - Comparative study of circulating immune complexes quantity detection by three assays--CIF-ELISA, C1q-ELISA and anti-C3 ELISA. AB - The assessment of the soluble immune complexes (IC) in human sera is traditionally performed by the C1q binding assay. In the present study, a novel method for the quantity of immune complexes was reported. The methodology was based on measuring their deposition on solid-phase C3 binding glycoprotein (CIF), using an enzyme-linked immunosorbent assay. We also used ELISA that employed anti C3 antibodies to determined the quantity of immune complexes. The three assays were evaluated for their performance characteristics on the same specially prepared samples: 55 normal sera, 99 sera from RA, 88 sera from SLE, and 27 sera from PSS. The results were compared by reference to a common standard-heat aggregated IgG that possesses many activities of immune complexes. Three of the tests used displayed almost the same specificity (over 95%), while their relative sensitivity varied depending on the disease sera tested. The sensitivity of the assays used was recorded highest for C1q ELISA-28.97% of positive sera, followed by CIF-ELISA-19.63% and lowest for anti-C3 ELISA-17.29%. A well-expressed correlation was found between CIF-ELISA and anti-C3 ELISA data (r=0.42), and a week correlation was noted when comparing CIF-ELISA and C1q ELISA IC levels detected (r=0.28). When the correlation coefficients were calculated individually for each disease category, they were clearly different, and that reflected indirectly in different sensitivities of the test for various disease categories. We also found that the results from the simultaneous performance of the tests demonstrated low percentage positive results when three or two assays were used. This is most probably due to the different assay abilities to detect IC with different sizes and composition, which shows that a small part of IC in the tested sera can be detected simultaneously by more than one assay. On the basis of the results obtained, we concluded that optimal screening for IC could be achieved by parallel application of several different methods. PMID- 11384666 TI - Reference values for peripheral blood B-lymphocyte subpopulations: a basis for multiparametric immunophenotyping of abnormal lymphocytes. AB - BACKGROUND AND OBJECTIVES: Immunophenotyping has become a useful tool for the differential diagnosis of chronic B-cell lymphoproliferative disorders. The aim of this work was to determine reference values of normal B-cell subpopulations. MATERIAL AND METHODS: Blood samples from 38 healthy volunteers were analyzed by multidimensional flow cytometry, using a panel of directly conjugated antibodies. Results were expressed as percent of positive B cells and as median fluorescence intensity, an indirect assessment of the expression level. RESULTS: CD20, CD22, CD24, CD40, CD79a, CD79b, FMC7, CD11a, CD18, CD44 were positive in the whole B cell population, whereas CD10, CD86, CD103, CD154 and FasL were almost absent from the B-lymphocyte population. 75% were IgD positive. The kappa/lambda ratio was 1.5. CD5, CD23, CD25, CD38, CD43, CD54, CD62L, CD80 and CD95 were positive in different B-cell subpopulations. The utility of all these markers in the differential diagnosis of chronic B-cell lymphoproliferative disorders is discussed. CONCLUSION: In order to interpret a pathological immunophenotype, it is necessary to refer to quantitative and qualitative values of normal B-cell subpopulations. PMID- 11384667 TI - Simultaneous flow cytometric measurement of K-562 megakaryocytic differentiation and CD56+ large granular lymphocyte cytotoxicity. AB - K-562 cells have the capacity to undergo multi-lineage differentiation, which may be crucial to their ability to serve as target reservoirs for CD56+ large granular lymphocytes (LGL). Conventional techniques using chromium release assays to measure lymphocyte-mediated cytotoxicity suffer from disadvantages, including radioactive contamination and the inability to simultaneously determine K-562 and/or CD56+ lymphocyte phenotypes. We illustrate here a three-color flow cytometric method providing for the simultaneous evaluation of K-562-CD56+ LGL binding, K-562 cell viability, and the status of K-562 cell differentiation. Phorbol 12-myristate 13-acetate (PMA) engenders megakaryocytic differentiation in K-562 cell populations, as measured by presentation of the beta(3) integrin (gpIIIa, CD61), while maintaining a negative expression of MHC-I and MHC-II molecules. Using the auto-fluorescence of K-562 cells, flow cytometry can be used to demonstrate a significant decrease in CD56+ LGL activity against K-562 cells in populations pre-incubated with PMA. The capacity of three-color flow cytometry to measure lymphocyte-target cell binding and cell death kinetics, while simultaneously determining target cell phenotype, permits the specific localization of CD61-expressing K-562 cells to areas inconsistent with CD56+ LGL mediated patterns of lysis. PMID- 11384668 TI - Extensive and long-term ex vivo production of dendritic cells from CD34 positive umbilical cord blood or bone marrow cells by novel culture system using mouse stroma. AB - We previously developed a system using murine strome (HESS-5), which could expand umbilical cord blood (UCB) stem and progenitor cells, especially CD34+/38- cells, in the presence of human recombinant cytokines. In this study, the ability of expanded UCB- or bone marrow (BM)-CD34+ cells to differentiate into dendritic cells (DCs) was examined. DCs could be induced either from short or long term cultured CD34+ cells after switching the cytokines from Flk-2/Flt-3 ligand, stem cell factor (SCF), thrombopoietin (TPO) to granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) (immature type) plus tumor necrosis factor (TNF)-alpha with stimulation by CD40L transfectant (mature type). Each immature or mature UCB-DCs showed a dextran uptake or a potent allo-T lymphocytes proliferative ability, respectively. Furthermore, those DCs from BM significantly stimulated auto-T lymphocytes in an antigen (varicella zoster virus) specific manner. In conclusion, a novel culture system using HESS-5 is useful to support a rapid and sustained generation of primitive myeloid cells which can develop into functional DCs. PMID- 11384669 TI - Cell biotinylation provides a sensitive and effective detection technique for cellular adhesion assays: comparison with existing methods. AB - A simple, sensitive, colorimetric labelling method was devised to quantify cell adhesion, based on labelling the cell plasma membrane with biotin. This method was applied in adhesion assays, which involved the adherence of biotin-labelled, PMA-stimulated, U937 cells. These cells resemble monocytes, and were bound onto fibronectin-coated wells and to an ECV304 cell monolayer. The adherent U937 cells were detected by the addition of a peroxidase-conjugated anti-biotin antibody and a soluble colorimetric substrate. This assay is convenient, fast and sensitive, and able to detect 320-1000 U937 cells under the conditions described. This study has used titration assays to compare the biotinylation method with the existing cell quantification approaches of 51Cr radiolabelling and antibody dependent ELISA. Chromium labelling was the most sensitive technique, but we found the biotinylation method to be more convenient than radioactive labelling and more sensitive than conventional ELISA. Biotinylated cells were also used very effectively in a Stamper-Woodruff adhesion assay with U937 cells binding to histological sections of atherosclerotic plaques. The selective detection of the bound cells permitted automated quantitation by image analysis. Whole cell biotinylation may have wider applications in biological research. PMID- 11384670 TI - Measurement of NK activity by the microcytotoxicity assay (MCA): a new application for an old assay. AB - Natural killer (NK) cells are spontaneously cytotoxic immune effector cells with the ability to selectively destroy tumor cells without harming normal cells. To perform this function, NK cells utilize two main cytotoxicity pathways, the well known perforin/granzyme-mediated secretory/necrotic killing and the recently defined TNF family ligand-mediated non-secretory/apoptotic killing. The former mechanism is manifested mainly against a few cultured leukemia cell targets, while the latter mediates killing against a large variety of tumor cell targets. Therefore, the biological role and significance of these mechanisms might be different. The NK cell-mediated necrotic killing has been reliably and selectively measured in humans by the standard 4-h 51Cr release assay (CRA) against K562 myeloid leukemia cell targets. However, no standardized high throughput assay is available for testing the NK cell-mediated apoptotic killing. Here, we introduce the modified MCA as a convenient method for measuring perforin/granzyme-independent NK cell-mediated apoptotic killing. The assay is performed in microwells of Terasaki tissue culture microtest plates, using adherent tumor cell targets, which are selectively susceptible to non secretory/apoptotic killing and resistant to secretory/necrotic killing mediated by NK cells. Target cells are plated in microwells and incubated overnight to adhere to the plastic surface and to regenerate cell surface-bound TNF family receptors. Following this adherence, target cells are co-incubated with freshly isolated human peripheral blood mononuclear leukocytes (PBMNL) or purified subpopulations of immune cells for 24 h in various effector/target (E/T) ratios. During this incubation, dead target cells become non-adherent and are removed by washing the wells. Remaining adherent (viable) target cells are fixed, stained and optically counted. A notable dose-dependent (peak at 200:1 E/T ratio), time dependent (peak at 24 h of incubation) and donor-dependent killing of tumor cells was consistently and reproducibly induced by PBMNL of normal donors. Using purified subpopulations of immune cells, it was demonstrated that among PBMNL, CD3(-)CD56(+)CD16(+) mature NK cells are the only mediators of tumor cell killing in MCA, as well as in CRA. Comparative studies of NK activity detected by MCA and CRA, performed with PBMNL from normal individuals and breast cancer patients, showed no significant correlation between the cytotoxicities measured in the two assays. In addition, while NK activity measured in CRA was normal in most breast cancer patients, NK activity assessed in MCA was decreased in a large majority of the patients. Thus, MCA is a sensitive NK assay, which is biologically different from CRA, and may be clinically relevant. MCA has also a higher throughput, and is more practical and economical than CRA. PMID- 11384671 TI - Quantification of ovine cytokine gene expression by a competitive RT-PCR method. AB - A quantitative competitive RT-PCR method was developed in order to measure IL 1beta, IL-4, IL-12, IFNgamma, TNFalpha and G(3)PDH mRNA from samples of ovine tissue such as lymph node or spleen. The main advantage of the method relies on the use, for each target sequence, of an internal competitor construct similar to the relevant target, but 4-bp different in size. This competitive strategy is validated by the equivalence of the amplification process, observed separately between competitor DNA and target DNA species. Furthermore, the copy number of each cytokine cDNA is normalized to a fixed copy number of G(3)PDH cDNA. The cDNA level of this constitutive gene was effectively shown to remain constant whatever the tissue studied and independently of the experimental conditions used. The accurate and reproducible data obtained permit the application of this quantitative RT-PCR method to measure the sheep cytokine response to Salmonella infection. Early induction of IFNgamma mRNA was observed in the draining lymph node 1 day after infection. At the same time, a strong increase of IL-1beta mRNA was observed in local and systemic lymphoid organs, suggesting the initiation of the inflammatory response. Finally, the overall results demonstrate the efficiency of the method and its suitability for further studies of the immune response in the ovine species. PMID- 11384672 TI - Flow cytometric quantitation of calcium-dependent and -independent mitogen stimulation of T cell functions in whole blood: inhibition by immunosuppressive drugs in vitro. AB - We have optimized assays to measure mitogen-stimulated rat lymphocyte activation in whole blood and have used these assays to quantitate the potencies of immunosuppressive drugs with different mechanisms of action. To define the optimal conditions for measuring T cell functions in whole blood, the effects of different concentrations of mitogens that activate T cells through calcium dependent and -independent pathways were measured over time. Proliferation was measured by tritium-labeled thymidine ([3H]-TdR) incorporation and by flow cytometric analysis of proliferating cell nuclear antigen (PCNA)/DNA content. Furthermore, we detected the increases in percent expression of cell-surface activation antigens (CD25, CD134, CD71, CD11a and CD54). Concanavalin A (Con A) stimulated maximum lymphocyte proliferation and expression of T cell surface activations by 72-96 h, which was 48 h later than stimulation by phorbol 12 myristate 13-acetate (PMA) plus anti-CD28 monoclonal antibody (mAb) or PMA plus ionomycin (IONO). Addition of sirolimus, tacrolimus, cyclosporine or the active metabolite of leflunomide, A77 1726, to mitogen-stimulated whole blood produced drug concentration-dependent inhibitions of lymphocyte proliferation and expression of cell surface activation antigen expression. From these data, we determined drug potencies (inhibitory concentration of 50%, IC(50)) and drug concentrations causing maximum inhibition of T cell functions (I(max)). We developed simple and reproducible assays to measure different lymphocyte functions in whole blood cultures. These assays were used to investigate the mechanisms of different immunosuppressive drugs. These methods can be exploited to measure T cell functions in blood collected from subjects treated with immunosuppressants in vivo. PMID- 11384673 TI - Novel centrifugal method for simple and highly efficient adenovirus-mediated green fluorescence protein gene transduction into human monocyte-derived dendritic cells. AB - Dendritic cells (DC) are professional antigen-presenting cells in the immune system. Gene transduction of DC with tumor-associated antigen (TAA) or other genes that enhance the immune reaction has been considered theoretically useful for DC-based immunotherapy. However, gene transduction of DC generated from human peripheral blood monocytes has been difficult due to its low efficiency, even when adenoviral vector was used at high multiplicity of infection (MOI). In the present study, we examined the effect of centrifugal force to enhance efficiency of adenovirus-mediated gene transduction into human monocyte-derived DC at various rotor speeds at various temperatures for various times. We judged the transduction efficiency using enhanced green fluorescence protein (EGFP) expressing adenoviral vector, and the best condition for centrifugal transduction was determined as 2000 x g at 37 degrees C for 2 h at an MOI of 10 or greater. At an MOI of 50 without centrifugation, the gene transduction efficiency was about 66% and mean fluorescence intensity (MFI) of EGFP expression was about 150 (at 37 degrees C for 2 h). With centrifugal transduction (2000 x g at an MOI of 50 at 37 degrees C for 2 h), 86% or more DC were gene-modified, and especially, MFI of EGFP expression was highly enhanced (MFI: about 3100 or greater). Centrifugally gene-transduced DC were not damaged and were thoroughly functional as measured by mixed lymphocyte reaction (MLR). The centrifugal method was also applicable to human monocytes and K562 cells. The centrifugal transduction method with adenoviral vector might be helpful for the generation of gene-modified DC. PMID- 11384674 TI - Double-blotting: a solution to the problem of non-specific binding of secondary antibodies in immunoblotting procedures. AB - "Double-blotting" (DB) was developed to overcome the problem of non-specific binding of secondary antibodies in immunoblotting (IB). After it had been probed by the primary antibody, the membrane with the blotted proteins was assembled with a second blank membrane and submitted to a second blotting under acidic conditions. The primary antibody molecules were thus desorbed from their corresponding antigen and transferred onto the second membrane, whereas the antigen and the interfering proteins remained bound to the first one. The second membrane could then be probed by the secondary antibodies without the risk of non specific binding. This method was developed for the study of erythropoietin (EPO) in concentrated urine since a strong non-specific binding of biotinylated secondary antibodies to some urinary proteins had been observed using classical IB protocols. PMID- 11384675 TI - Enzyme-linked immunosorbent assay (ELISA) for the specific detection of apoptotic cells and its application to rapid drug screening. AB - We have developed a solid-phase ELISA for the specific and sensitive detection of apoptotic cells. This method is based on the ability of a monoclonal antibody (MAb) against single-stranded DNA (ssDNA) to specifically identify apoptotic cells. The assay involves binding of cells to 96-well microtiter plates, treatment of the attached cells with formamide to denature DNA in apoptotic cells and one-step staining of the denatured DNA with a mixture of anti-ssDNA MAb and peroxidase-conjugated anti-mouse IgM. A near linear increase in signal was seen as the number of apoptotic cells increased from 500 to 5000. Untreated and necrotic cells or cells with single-stranded DNA breaks induced by H(2)O(2) did not produce signal above the background level. In leukemic cell cultures grown, treated with ID(50) concentration of etoposide, stained and analyzed in the same 96-well assay plate, intense ELISA signal was detected. The ratio of absorbance values from drug resistant and drug-sensitive cell lines treated with etoposide was in agreement with the degree of resistance determined by growth inhibition assays. These data show that this ELISA has sufficient sensitivity for use in drug screening protocols. In breast cancer cell cultures treated with cisplatin, ELISA absorbance increased only after treatment with drug concentrations 10-fold higher than concentrations inducing 95% growth inhibition. In cultures treated with staurosporine, there was a near linear relation between the ELISA absorbance values and cytotoxicity in the range of 15-92% growth inhibition. The absence of apoptotic signal in breast cancer cells treated with cytotoxic concentrations of cisplatin indicated that this drug kills cells by non-apoptotic mechanisms, whereas apoptosis was the dominant mechanism of cell death caused by staurosporine. The formamide-MAb apoptosis ELISA described here may provide a basis for high-throughput screening of drugs based on their ability to induce or suppress apoptosis. PMID- 11384676 TI - Flow cytometric DNA-quantification of three-color immunophenotyped cells for subpopulation specific determination of aneuploidy and proliferation. AB - A method is described for three-color immunophenotyping and simultaneous DNA quantification using a flow cytometer equipped with a 488-nm argon laser and a mercury lamp (UV). The approach includes reproducible immunophenotyping comparing antigen expression before and after cell manipulation for DNA-measurement. The coefficients of variation after DNA-staining (CV=3.13 for T-cells in peripheral blood and CV=3.38 for T-cells in bone marrow) were adequate for exact DNA analysis. For aneuploidy detection, a true internal standard was established measuring, for example, the DNA-content of T-cells in B-cell disease simultaneously with the DNA-content of the malignant cells. Using this method, aneuploidies could be unequivocally detected in 17 out of 24 patients with multiple myeloma. Furthermore, intratumor heterogeneities in DNA-content and antigen expression could be recognized, allowing an exact separation of tumor cells and normal hematopoiesis. The study also demonstrated the importance of exact immunophenotypic characterization of lymphocyte subpopulations and the determination of their specific proliferation, for example after proliferation induction in cell cultures. Future studies should address the applicability of this rather simple multiparameter approach for simultaneous immunophenotyping and DNA-measurement especially in the detection of minimal amounts of aneuploid cells after chemotherapy. PMID- 11384677 TI - A generic capture ELISA for recombinant proteins fused to glutathione S transferase: validation for HPV serology. AB - An enzyme-linked immunosorbent assay (ELISA) system has been developed that uses glutathione crosslinked to casein as capture protein to bind recombinant protein antigens fused to N-terminal glutathione S-transferase (GST). The method allows simple and efficient immobilization and one-step purification of overexpressed recombinant antigens from crude lysates on ELISA plates coated with glutathione casein. Several antigens can be tested in parallel under the same conditions without the need to biochemically purify or renature the proteins. An additional undecapeptide epitope fused to the C-terminus of each antigen permits the detection and quantification of any full-length protein antigen bound to the ELISA plate with one single monoclonal antibody. The ELISA system was applied with four antigens to detect antibodies against E6 and E7 proteins of human papillomavirus types 16 and 18. Antibody reactivities of 164 sera from patients with cervical carcinoma and healthy individuals were in good agreement with those determined using a previously established capture ELISA with biochemically purified and renatured proteins as antigens although the GST capture ELISA was more sensitive with no loss of specificity. The GST capture ELISA could be adapted to provide standardized antibody assays for many protein antigens. PMID- 11384678 TI - A method that allows easy characterization of tumor-infiltrating lymphocytes. AB - A method was developed to compare the lymphocytic infiltrates in regressing vs. progressing experimental mouse tumors using a model for human papillomavirus-16 (HPV-16) oncoprotein-linked cancer. Tumor cells mixed with matrigel, composed of natural matrix substances that provide a basement membrane structure for adherent cells, were inoculated into mice vaccinated with an efficacious vaccine to the E7 oncoprotein or a vaccine to a control antigen. The tumor cells remained within the solidified gel and recruited a cellular infiltrate that could readily be analyzed upon removal of the gelatinous mass containing progressing or regressing tumors. The results show that tumors recruit activated CD8(+) T cells regardless of their antigen specificity. In regressing tumors expressing an appropriate target antigen for the vaccine-induced CD8(+) T cells, a strong increase of the tumor antigen-specific T cell population was observed over time. Progressing tumors that lacked the target antigen for the activated CD8(+) T cell population did not show this selective enrichment. PMID- 11384679 TI - A novel flow cytometric assay for quantitation and multiparametric characterization of cell-mediated cytotoxicity. AB - Cell-mediated cytotoxicity is a crucial mechanism involved in several fundamental immunological processes such as protection against intracellular pathogens or termination of an immune response. This phenomenon is classically evaluated by the 51Cr release assay, which requires a radioactive isotope and does not permit the characterization of cells involved in the cytotoxic reaction. We describe a new flow cytometry method, developed in the context of CD95-mediated cell death, which allows the precise quantitation of cell-mediated cytotoxicity and the detection of intracellular events involved in the cytotoxic process. This assay uses a combination of two dyes, i.e. 5- (and 6-) carboxyfluorescein diacetate succinimydyl ester (CFSE) to label effector cells and 7-amino actinomycin D (7 AAD) to stain apoptotic target cells. We show that this assay is more sensitive than the 51Cr release assay and makes it possible to quantitate the percentage of cell lysis and, concomitantly, to immunophenotype target cells. It also facilitates the analysis of some events of the apoptotic pathway such as caspase activation or the expression of mitochondrial molecules. This new assay should contribute to a better understanding of the mechanisms involved in cell-mediated cytotoxicity in normal and pathological situations. PMID- 11384680 TI - The use of 7-amino-actinomycin D in the analysis of Candida albicans phagocytosis and opsonization. AB - We describe the use of 7-amino-actinomycin D (7AAD) to measure phagocytosis and the opsonizing capacity of serum. Heat-inactivated Candida albicans was previously stained with 7AAD and incubated with resident peritoneal macrophages. The samples were analyzed by flow cytometry and phagocytic cells were identified by their bright red fluorescence. This is a rapid, reproducible and reliable one step procedure and provides a means of evaluating low levels of phagocytosis. PMID- 11384681 TI - Mammalian expression and hollow fiber bioreactor production of recombinant anti CEA diabody and minibody for clinical applications. AB - Genetically engineered radiolabeled antibody fragments have shown great promise for the radioimmunoscintigraphy of cancer. Retaining the exquisite specificity of monoclonal antibodies yet smaller in molecular size, antibody fragments display rapid tumor targeting and blood clearance, a more uniform distribution in the tumor, and present a lower potential to elicit an immune response. However, one of the factors that has limited clinical evaluation of these antibody-derived proteins has been the difficulty in expressing and purifying the quantities necessary for clinical trials. This study outlines the capability of mammalian expression for the production of recombinant antibody fragments intended for clinical use. Two anti-carcinoembryonic antigen antibody fragments, the T84.66/212 Flex minibody (scFv-C(H)3) and the T84.66 diabody (scFv dimer) have been previously expressed and have shown excellent radioimaging properties in tumor bearing animals. To proceed toward human studies, these high affinity recombinant fragments and a second minibody version, the T84.66/GS18 Flex minibody, were expressed using a high-level mammalian expression system. Production of all three antibody fragments in a small-scale hollow fiber bioreactor resulted in 137-307 mg of crude antibody harvest. A purification protocol that employed ceramic hydroxyapatite and anion exchange chromatography resulted in 50-150 mg of purified T84.66 diabody and T84.66 minibody. The development of this level of research grade material established conditions for clinical production as well as provided material to complete pre-clinical studies and undertake protein crystallization studies. Scale-up for clinical studies produced 3.4 g of the T84.66 minibody in the harvest. A portion of this material was purified yielding 180 mg of highly purified T84.66 minibody intended for pilot radioimmunoscintigraphy studies of carcinoembryonic antigen (CEA) positive disease. PMID- 11384682 TI - Protocols for high efficiency, stage-specific retroviral transduction of murine fetal thymocytes and thymic epithelial cells. AB - Viral vectors have the potential to provide a fast and economic alternative to transgenic methods for manipulating gene expression in studies of immune system development and function. Although protocols exist for the infection of hematopoietic precursors and peripheral T cells in vitro, critical stages of T cell differentiation are strictly dependent upon a three-dimensional thymic architecture and their analysis poses unique technical challenges. Whole fetal thymic lobes have been used as targets for retroviral and adenoviral infection, both in situ and in vitro, but this approach does not allow for discrimination between lymphoid and stromal components. Isolated thymocytes have been infected by co-culture with viral producer cells, but under these conditions they rapidly lose their developmental potential. To overcome these problems we have combined a number of efficient techniques for retroviral production, concentration, and infection that allow us to rapidly achieve significant transduction rates of purified populations of double-negative (DN) and double-positive (DP) thymocytes, single-positive (SP) T lymphocytes, as well as fetal thymic MHC II(+) epithelial cells without the need for co-culture with viral producer cells. Reaggregate thymic organ culture (RTOC) techniques were used to assess the development and function of transduced cells in defined cellular environments. As a demonstration of the utility of these methods, CD80 (B7.1) was transduced into thymic epithelial cells and shown to allow them to mediate negative selection of DP thymocytes, and to act as antigen-presenting cells (APC) to mature T cells. The ability to genetically manipulate primary cells of a specified type and differentiation stage provides a powerful complement to RTOC techniques for the study of T cell development. PMID- 11384683 TI - A high-capacity alkaline phosphatase reporter system for the rapid analysis of specificity and relative affinity of peptides from phage-display libraries. AB - We describe a novel reporter enzyme cassette system which enables the analysis of large numbers of linear and cyclic peptides in terms of their binding to a specific target molecule. In this system, peptides selected for target binding from random peptide phage-display libraries are expressed as cloned fusion proteins with bacterial alkaline phosphatase. The binding specificity and relative affinity of each peptide-enzyme fusion protein is then evaluated in a target-specific ELISA. This strategy enables direct identification of the highest affinity peptides, specific for a given target, which can then be sequenced at the DNA level to derive their peptide sequences. This eliminates the need to sequence large numbers of clones to establish consensus sequences for binding peptides. This approach also eliminates the need for peptide synthesis or phage ELISA to determine relative binding affinities, which can be technically difficult. Identification of binding peptides based on specificity and relative affinity, rather than conforming to an amino acid consensus sequence, enables the rapid evaluation of hundreds of candidate peptides and identification of rare (non-consensus) binding peptides which may otherwise be missed. PMID- 11384684 TI - Antibodies in haystacks: how selection strategy influences the outcome of selection from molecular diversity libraries. AB - Antibodies against most antigens can be isolated from high quality phage antibody libraries. However, not all antibodies binding a particular antigen are necessarily found when standard selections are performed. Here we investigate the effect of two different selection strategies on the isolation of antibodies against a number of different antigens, and find that these different strategies tend to select different antibodies, with little overlap between them. This indicates that the full diversity of these libraries is not tapped by a single selection strategy and that each selection strategy imposes different selective criteria in addition to that of antigen binding. To fully exploit such libraries, therefore, many different selection strategies should probably be employed for each antigen. The use of alternative strategies should be considered when selection apparently fails, or when the number of different antibodies recognizing an antigen needs to be maximised. Furthermore, the microtitre selection strategy developed is likely to prove useful in the application of phage antibody libraries to the human genome project, allowing the high throughput selection of antibodies against multiple antigens simultaneously. PMID- 11384685 TI - An immunohistochemical method for the detection of proteins in the vertebrate lens. AB - Fluorescence immunohistochemistry has traditionally been difficult or impossible to perform on the vertebrate lens because of its extremely high protein content. Described here is a robust and rapid method for preparing and labeling vertebrate eyes for confocal microscopy. This technique has successfully been applied to localize proteins in the lens epithelium and capsule, as well as the primary and secondary fibers. This technique preserves tissue morphology and coupled with double and triple labeling, has allowed localization of proteins bound to plasma membrane, basement membrane, nucleus, endoplasmic reticulum as well as sub nuclear compartments. In addition, the present technique has proven useful for fluorescent immunohistochemical analysis of diverse tissues including whole embryos, adult muscle, pancreas, and liver. This procedure allowed us to successfully localize a wide variety of antigens on diverse vertebrate tissues including the more challenging vertebrate lens. PMID- 11384686 TI - A virus binding assay for studying the antigenic landscape on intact, native, primary human immunodeficiency virus-type 1. AB - This protocol describes a simple assay that can be used to study the nature of exposure of antigenic epitopes and antigenic relatedness of different intact, native HIV-1 strains. The assay is based on the principle that mAbs coated on microtiter wells bind to epitopes on the surface of intact, native virions. The bound virion is then lysed to release p24, which is then quantitated (pg/ml) to give a measure of the amount of virion bound to the mAb. High p24 levels released after lysis correlate with high level capture of virions by mAbs, and as such, reflect good exposure of the epitope on the virion. Likewise, binding patterns of a specific mAb with different virus strains reveal information on their antigenic relatedness. In establishing this assay, the nature of exposure of antigenic epitopes and the antigenic relatedness of six intact, native HIV-1 virions of clades A, B, C, D, F and G were examined using anti-HIV-1 mAbs directed at epitopes in the V2, V3, CD4bd and C5 of gp120, and in clusters I and II of the gp41 region. Analysis of the binding data shows that mAbs directed at epitopes in the V3, C5 and gp41 Cluster I region bound best to the viruses examined, suggesting that these are the regions most exposed and conserved on intact, native HIV-1 virions of different clades. Epitopes in the V2 and CD4bd of gp120, and in gp41 cluster II, are not exposed on intact, native virions. PMID- 11384687 TI - Vasopressin during abdominal hysterectomy: a randomized controlled trial. AB - OBJECTIVE: To investigate if the use of vasopressin during abdominal hysterectomy would decrease blood loss. METHODS: Fifty-one patients undergoing abdominal hysterectomy with the diagnosis of leiomyomatous uterus were randomized and received either vasopressin 10 units/10 mL of normal saline or 10 mL of normal saline, injected 5 mL bilaterally, 1 cm medial to the uterine vessels into the lower uterine segment. The sample size was determined assuming a one-third reduction in total blood loss would be clinically relevant. A power analysis determined that 25 patients would be required in each group to assure a power of 0.80, at the.05 significance level. RESULTS: Overall, the two groups were very similar with regard to their demographics, preoperative diagnosis, and relevant findings at the time of surgery. The mean total blood loss in the vasopressin and placebo groups was 445.41 mL and 748.42 mL, respectively. Total blood loss was significantly decreased by 40% in the vasopressin group compared with the placebo group (P <.001). There was no statistically significant difference between the two groups with respect to possible confounding variables or surgical complications. CONCLUSION: Injection of vasopressin into the uterus at the time of abdominal hysterectomy significantly reduces blood loss without increasing morbidity. We have shown that it is a useful adjunct during abdominal hysterectomy. PMID- 11384688 TI - Uterosacral ligament: description of anatomic relationships to optimize surgical safety. AB - OBJECTIVE: To determine the optimal site in the uterosacral ligament for suspension of the vaginal vault with regard to adjacent anatomy and suspension strength. METHODS: Fifteen female cadavers were evaluated between December 1998 and September 1999. Eleven hemisected pelves were dissected to better define the uterosacral ligament and identify adjacent anatomy. Ureteral pressure profiles with and without relaxing incisions were done on four fresh specimens. Suture pullout strengths also were assessed in the uterosacral ligament. RESULTS: The uterosacral ligament was attached broadly to the first, second, and third sacral vertebrae, and variably to the fourth sacral vertebrae. The intermediate portion of the uterosacral ligament had fewer vital, subjacent structures. The mean +/- standard deviation distance from ureter to uterosacral ligament was 0.9 +/- 0.4, 2.3 +/- 0.9, and 4.1 +/- 0.6 cm in the cervical, intermediate, and sacral portions of the uterosacral ligament, respectively. The distance from the ischial spine to the ureter was 4.9 +/- 2.0 cm. The ischial spine was consistently beneath the intermediate portion but variable in location beneath the breadth of the ligament. Uterosacral ligament tension was transmitted to the ureter, most notably near the cervix. The cervical and intermediate portions of the uterosacral ligament supported more than 17 kg of weight before failure. CONCLUSION: Our findings suggest that the optimal site for fixation is the intermediate portion of the uterosacral ligament, 1 cm posterior to its most anterior palpable margin, with the ligament on tension. PMID- 11384689 TI - Hysterectomy in Veterans Affairs Medical Centers. AB - OBJECTIVE: To examine the indications and surgical morbidity for women veterans who underwent hysterectomies in Department of Veterans Affairs Medical Centers (VAs). METHODS: Data on hysterectomies performed in VAs from 1991 to 1997 were abstracted from a surgical quality improvement program. RESULTS: Records of 1722 women who had hysterectomies in VAs over 6 years were examined. Women were predominately white (62%) and their average age was 42.5 years. Operations included abdominal (74%), vaginal (22%), and laparoscopic-assisted (4%) methods. The most common indications for surgery included uterine leiomyomas (31%), abnormal uterine bleeding (14%), and endometriosis (11%). Indications differed by race (P <.01); nonwhite women were most likely to have surgery for leiomyoma (51%), whereas white women had hysterectomies for leiomyomas (19%), abnormal bleeding (15%), endometriosis (13%), and genital prolapse (11%). The mean postoperative stay was significantly longer for abdominal hysterectomies (4.51 days) than either vaginal or laparoscopic-assisted hysterectomies (2.92 and 2.21 days, respectively; P <.001). The overall complication rate within 30 days was 9%, and the most frequent complication was urinary tract infection (3.3%). CONCLUSION: Women who underwent hysterectomies in VAs had low complication rates, comparable to hysterectomy complication rates in the United States generally. PMID- 11384690 TI - Office screening test for intrinsic urethral sphincter deficiency: pediatric Foley catheter test. AB - OBJECTIVE: To evaluate an office-based test as a screening method for intrinsic sphincter deficiency. METHODS: One hundred seventy-three women with urinary incontinence were evaluated prospectively by complete urodynamic studies. After catheterization, the pediatric Foley catheter test was performed on an empty bladder by withdrawing the inflated bulb of an 8-French Foley catheter through the urethra. The test was considered positive if the inflated catheter bulb could be withdrawn completely through the urethra. Women with grade 3 genital prolapse or higher were excluded. Intrinsic sphincter deficiency was defined as the presence of genuine stress incontinence and low maximum urethral closure pressure (at most 20 cm H(2)O). RESULTS: Seventy-six of 173 women (44%) had positive tests and 97 (56%) had negative tests. Seventy-six percent of those with positive tests were diagnosed with intrinsic sphincter deficiency, compared with 19% in women with negative tests (P <.001). All women with positive tests and negative cotton swab tests had intrinsic sphincter deficiency. The sensitivity, specificity, and positive and negative predictive values for diagnosing intrinsic sphincter deficiency were 76, 81, 76, and 81%, respectively. CONCLUSION: A positive pediatric Foley catheter test in the absence of urethral mobility strongly suggests intrinsic sphincter deficiency. The pediatric Foley catheter test may be useful in screening for intrinsic sphincter deficiency. PMID- 11384691 TI - Microinvasive carcinoma of the cervix: site of first focus of invasion. AB - OBJECTIVE: To evaluate the topography of the first invasive focus in microinvasive squamous cell carcinoma of the uterine cervix. METHODS: We studied 120 formalin-fixed and paraffin-embedded cold-knife conization specimens processed as step-serial sections. Four types of microinvasion were distinguished according to site: type I (ectocervical outside the last cervical gland), type II (ectocervical between the external os and the last gland and in connection with the surface epithelium), type III (ectocervical between the external os and the last gland but deep in cervical glands), and type IV (intracervical and in connection with the surface epithelium). RESULTS: A total of 142 early invasive foci were seen in the 120 cones. A single focus was seen in 106 (88%) specimens, whereas 14 (12%) had more than one focus. The foci were classified as type I in 16 (11%), type II in 31 (22%), type III in 70 (49%), and type IV in 25 (18%) cases. CONCLUSION: One half of the early invasive foci originated at the surface epithelium (types I, II, IV), either at the ectocervix (types I, II) or in the endocervix (type IV). PMID- 11384692 TI - Calcitonin gene- and parathyroid hormone-related peptides in preeclampsia: effects of magnesium sulfate. AB - OBJECTIVE: To determine whether circulating levels of calcitonin gene-related peptide (CGRP) and parathyroid hormone-related peptide (PTHrP) are altered in preeclampsia, and to assess the effects of magnesium sulfate therapy on circulating levels of these two peptides. METHODS: The study population included 25 women with preeclampsia and 25 normotensive controls of similar gestational age. The effects of magnesium sulfate therapy were evaluated in 17 of the 25 preeclamptic women. Circulating levels of immunoreactive CGRP and PTHrP, including calcium, magnesium, and phosphate in the maternal and umbilical cord serum were measured. RESULTS: The frequency of preeclampsia subjects with nondetectable PTHrP (under 3 pg/mL) was significantly higher (92% versus 48%, P <.001), whereas maternal serum CGRP levels were significantly lower (50 +/- 19 versus 90 +/- 23 pg/mL, P <.001). Similarly, the frequency of newborns with nondetectable PTHrP levels in umbilical serum was significantly higher (68% versus 36%, P <.05), whereas the levels of CGRP were significantly lower (67 +/- 17 versus 79 +/- 16 pg/mL, P <.05). Magnesium sulfate treatment resulted in a significant increase in maternal circulating CGRP levels (64 +/- 17 versus 47 +/- 18 pg/mL, P <.05) with no changes in PTHrP. CONCLUSION: Maternal circulating PTHrP and CGRP concentrations were significantly lower in women with preeclampsia, which may contribute to the development and maintenance of hypertension during pregnancy. Furthermore, magnesium sulfate therapy increased the levels of CGRP in the maternal circulation. PMID- 11384693 TI - Decreased maternal serum placenta growth factor in early second trimester and preeclampsia. AB - OBJECTIVE: To compare early second-trimester maternal serum placenta growth factor concentrations in patients with subsequent development of preeclampsia and those with normal pregnancies. METHODS: We conducted a case-control analysis of stored maternal serum of 27 women who subsequently developed preeclampsia and 227 randomly selected normal controls during the gestational period of 14-19 weeks. Using such a sample size, there was a greater than 95% power to test a difference in the primary study interest. A quantitative sandwich enzyme immunoassay was used to measure the maternal serum placenta growth factor concentration. For statistical analysis, Mann-Whitney U tests, multiple linear regression analysis, multivariable logistic regression model, and receiver-operating characteristic (ROC) curve were used. P <.05 was considered statistically significant. RESULTS: Maternal serum placenta growth factor concentration was associated with the occurrence of subsequent preeclampsia (P <.001) and gestational age (P <.001). The median (interquartile range) of multiples (MoM) of the gestational age stratified median for placenta growth factor in preeclampsia was 0.55 (0.33, 0.85). The ROC curve revealed that the specificity was 70% when the diagnostic sensitivity was 70%, and the optimal cutoff value of placenta growth factor MoM was 0.76. The risk of developing preeclampsia subsequently was increased 2.5-fold for maternal serum placenta growth factor concentration decrements of 0.1 MoM. CONCLUSION: A decreased maternal serum placenta growth factor concentration in the early second trimester is highly associated with the subsequent development of preeclampsia, but a large prospective study is needed to explore its use as an early predictor for the condition. PMID- 11384694 TI - Relative glucose tolerance and subsequent development of hypertension in pregnancy. AB - OBJECTIVE: To test the hypothesis that relative carbohydrate tolerance, an indicator of insulin resistance, predicts subsequent risk for hypertension of pregnancy among previously normoglycemic, normotensive women. METHODS: We conducted a nested case-control study in women enrolled at a large Colorado urban health maintenance organization. Subjects were previously healthy pregnant women who tested abnormal on their initial 50-g glucose screens and subsequently completed 3-hour, 100-g oral glucose tolerance tests. Cases were 54 previously normotensive women who subsequently developed hypertension and controls were 51 subjects with normotensive pregnancies, matched to cases on parity. Subjects diagnosed with gestational diabetes (17 cases, six controls) were excluded from the main analyses. RESULTS: Among the 82 normoglycemic women (45 controls, 37 cases, 13 preeclampsia, 24 gestational hypertension), mean post-load glucose levels and total glucose area under the curve were significantly higher in cases than in controls (P < or =.04) and were positively correlated with peak mean arterial pressure. After adjustment for potential confounders, 2-hour post-load glucose levels remained strongly related to risk for hypertension (adjusted odds ratios = 1.48; 95% confidence interval 1.13, 1.92, per 10 mg/dL increase) and to peak mean arterial blood pressure (r =.23, P =.04), as did total glucose area under the curve (P < or =.04). Cases were also more likely to have had one abnormal glucose tolerance test (28% versus 5%, P =.004). Stratifying analyses by case severity (preeclampsia and gestational hypertension) yielded similar results. Among all subjects, more cases than controls were also diagnosed with gestational diabetes (31% versus 12%, P =.008). CONCLUSION: These findings are consistent with the hypothesis that insulin resistance precedes the clinical onset of hypertension in pregnancy, and may be important in the etiology of hypertension. PMID- 11384695 TI - Prolonged pregnancy: induction of labor and cesarean births. AB - OBJECTIVE: To determine the effects of labor induction on cesarean delivery in post-date pregnancies. MATERIALS AND METHODS: A total of 1325 women who reached 41 weeks' gestation between December 1, 1997, and April 4, 2000, and who were scheduled for induction of labor at 42 weeks were included in this prospective observational study. Cesarean delivery rates were compared between those women who entered spontaneous labor and those who underwent induction. Women with any medical or obstetric risk factors were excluded. A power analysis was performed to determine how many patients would be required to show no effect of labor induction on cesarean delivery with a beta of.8 and an alpha of.05. Approximately 5200 patients would be required, taking an estimated 28 years to accrue at our institution. RESULTS: Admission to delivery was longer (5.7 compared with 11.1 hours, P =.001) and more likely to extend beyond 10 hours (55 compared with 24%, P =.001) in the induction group. Cesarean deliveries were increased in the induced group (19 compared with 14%, P <.001) due to cesarean for failure to progress (14 compared with 8%, P <.001). Independent risk factors for cesarean delivery included nulliparity, undilated cervix prior to labor, and epidural analgesia. Correction for these risk factors using logistic regression analysis revealed that it was the risk factors, and not induction of labor per se, that increased cesarean delivery. CONCLUSION: Risk factors intrinsic to the patient, rather than labor induction itself, are the cause of excess cesarean deliveries in women with prolonged pregnancies. PMID- 11384696 TI - Nuchal translucency and the acceptance of invasive prenatal chromosomal diagnosis in women aged 35 and older. AB - OBJECTIVE: To investigate the effect of nuchal translucency screening on use of prenatal diagnosis for chromosomal abnormalities in women aged 35 and older. METHODS: Two groups of women, referred to our center for prenatal karyotype diagnosis because of maternal age, were compared: one in 1995 and the other in 1999 after the introduction of nuchal translucency measurement. Each woman received nondirective genetic counseling, and for the 1999 group, nuchal translucency results were also discussed. Risks of transabdominal chorionic villi sampling (CVS) and amniocentesis, laboratory techniques, genetic results, and local experiences were discussed. Patient's decision to undergo prenatal diagnosis, acceptance of the nuchal translucency test (in the 1999 group), and the rate of chromosomal abnormalities diagnosed by transabdominal CVS and amniocentesis, were considered. RESULTS: Two hundred twenty-one of 982 (22%) women in the 1995 group and 421 of 1386 (30%) in the 1999 group, after nondirective genetic counseling declined invasive diagnosis (P <.05). In the 1999 cohort, 1088 of 1089 (99.9%) women of appropriate gestational age had nuchal translucency measurement. Among women seen in 1995, 214 opted for transabdominal CVS (31%) and 476 (69%) for amniocentesis. Nineteen abnormal karyotypes were detected, six by transabdominal CVS and 13 (68.5%) by amniocentesis. In 1999, 266 women (29%) opted for transabdominal CVS and 650 (71%) for amniocentesis. Twenty abnormal karyotypes were detected, 13 (65%) by transabdominal CVS and seven (35%) by amniocentesis (P <.05). CONCLUSION: Knowledge of nuchal translucency could lead to a decrease in the demand for invasive diagnosis and to a more frequent diagnosis by first-trimester transabdominal CVS. PMID- 11384697 TI - Indomethacin tocolysis and intraventricular hemorrhage. AB - OBJECTIVE: To determine the association between indomethacin tocolysis and neonatal intraventricular hemorrhage. METHODS: Fifty-six preterm neonates with intraventricular hemorrhage were matched by gestational age with neonates (n = 224) without this morbidity. Maternal and neonatal charts were reviewed to ascertain the type of tocolytic exposure experienced by the neonate. Other maternal and neonatal demographic and outcome data were also abstracted. Results were analyzed using the Student t test, chi(2) analysis, and multivariable logistic regression. The number of studied subjects provided 80% power to determine if antenatal exposure to indomethacin was twice as likely among infants with intraventricular hemorrhage. RESULTS: Univariate analysis revealed that there were no significant differences between the study and control groups with respect to maternal age, parity, or betamethasone exposure. Infants with intraventricular hemorrhage were significantly more likely to be born at an earlier gestational age, a lower birth weight, after maternal chorioamnionitis, after vaginal delivery, and after exposure to either indomethacin alone or a combination of indomethacin and magnesium. Additionally, their neonatal course was significantly more likely to be complicated by sepsis and respiratory distress syndrome. In a multivariable logistic model, only gestational age, chorioamnionitis, vaginal delivery, and respiratory distress syndrome continued to be significantly associated with intraventricular hemorrhage. Indomethacin exposure, either as single-agent (adjusted odds ratio 1.3, 95% confidence interval 0.5, 3.3) or combination tocolytic therapy (adjusted odds ratio 2.0, 95% confidence interval 0.8, 4.8), was not significantly associated with intraventricular hemorrhage. CONCLUSION: Indomethacin tocolysis is not associated with an increased risk of intraventricular hemorrhage. PMID- 11384698 TI - Excessive uterine activity accompanying induced labor. AB - OBJECTIVE: To estimate the incidence and timing of excessive uterine activity accompanying induction of labor with misoprostol using different routes (oral or vaginal) and forms (intact tablet or crushed) and to compare these with dinoprostone gel, oxytocin, and spontaneous labor. METHODS: This retrospective cohort study included 519 women at term who had labor induced and 86 women at term in spontaneous labor. Induction agents included misoprostol, dinoprostone, or oxytocin. Fetal heart rate and uterine activity tracings were analyzed independently by three maternal-fetal medicine physicians. The diagnosis of tachysystole or hyperstimulation required the agreement of two or more reviewers. RESULTS: The incidence of tachysystole was highest with misoprostol administered by vaginal tablet (misoprostol vaginal tablet 50 microg every 4 hours, 48.6%; vaginal tablet crushed 50 microg and suspended in hydroxyethyl gel every 4 hours, 30.7%, P =.009; oral tablet 50 microg every 4 hours, 22.2%, P =.001; oral tablet crushed 50 microg every 4 hours, 15.5%, P <.001; dinoprostone gel, 33.0%, P =.022; intravenous oxytocin, 30.2%, P =.027; and spontaneous onset of labor, 23.3%, P <.001). Hyperstimulation occurred more often with dinoprostone gel (16.5%) than with other forms of induction or spontaneous labor. Hyperstimulation occurred significantly more often with vaginal misoprostol crushed tablet (7.9%) and vaginal misoprostol intact tablet (7.6%) than with crushed oral misoprostol (1.0%) (P =.016 and.018, respectively). There was a shorter time to tachysystole with increasing doses of vaginal misoprostol tablet (P =.01). CONCLUSION: The incidence of tachysystole and hyperstimulation, and time to tachysystole, varied depending on the route and form of misoprostol given. PMID- 11384699 TI - Cost-effectiveness of a trial of labor after previous cesarean. AB - OBJECTIVE: To determine the cost-effective method of delivery, from society's perspective, in patients who have had a previous cesarean. METHODS: We completed an incremental cost-effectiveness analysis of a trial of labor relative to cesarean using a computerized model for a hypothetical 30-year old parturient. The model incorporated data from peer-reviewed studies, actual hospital costs, and utilities to quantify health-related quality of life. A threshold of $50,000 per quality-adjusted life-years was used to define cost-effective. RESULTS: The model was most sensitive to the probability of successful vaginal delivery. If the probability of successful vaginal birth after cesarean (VBAC) was less than 0.65, elective repeat cesarean was both less costly and more effective than a trial of labor. Between 0.65 and 0.74, elective repeat cesarean was cost effective (the cost-effectiveness ratio was less than $50,000 per quality adjusted life-years), because, although it cost more than VBAC, it was offset by improved outcomes. Between 0.74 and 0.76, trial of labor was cost-effective. If the probability of successful vaginal delivery exceeded 0.76, trial of labor became less costly and more effective. Costs associated with a moderately morbid neonatal outcome, as well as the probabilities of infant morbidity occurring, heavily impacted our results. CONCLUSION: The cost-effectiveness of VBAC depends on the likelihood of successful trial of labor. Our modeling suggests that a trial of labor is cost-effective if the probability of successful vaginal delivery is greater than 0.74. Improved algorithms are needed to more precisely estimate the likelihood that a patient with a previous cesarean will have a successful vaginal delivery. PMID- 11384700 TI - Changes in providers' views and practices about emergency contraception with education. AB - OBJECTIVE: To assess changes in the prescribing practices, knowledge, attitudes, and perceptions of health care providers after an educational program about emergency contraception. METHODS: Health care providers completed self administered questionnaires before and 1 year after full implementation of the project. The 102 providers who completed both questionnaires were physicians (64%) and mid-level professionals from 13 San Diego County Kaiser Permanente medical offices working in departments such as obstetrics and gynecology, primary care, and emergency medicine. RESULTS: The frequency of prescription for emergency contraceptive pills increased significantly from baseline to follow-up. There was an increase of almost 20% in the percentage who prescribed emergency contraception at least once a year. Knowledge also improved significantly, and perceptions of barriers to prescribing emergency contraceptive pills within the health maintenance organization decreased significantly. In contrast, attitudes about emergency contraception showed little change. CONCLUSION: This study suggests that providers who participate in in-service training and other aspects of a demonstration project show changes in perceptions, knowledge, and behavior. However, findings also suggest that significant gaps remain in knowledge about medications, side effects, and mode of action. It is likely that many providers in other health care settings also need additional information and training concerning protocols of emergency contraception provision and its modes of action and effects. PMID- 11384701 TI - Fetal syphilis: clinical and laboratory characteristics. AB - OBJECTIVE: To examine the pathophysiology of fetal syphilis and correlate hematologic, immunologic, and sonographic findings. METHODS: Twenty-four women with untreated syphilis during pregnancy were prospectively identified. Sonography with amniocentesis and percutaneous umbilical blood sampling were performed. Darkfield examination, rabbit infectivity testing, and polymerase chain reaction for detection of Treponema pallidum were performed on amniotic fluid. Hematologic and chemical testing of fetal blood was performed using standard techniques. Fetal antitreponemal IgM was detected by Western blot assay. Maternal syphilis was treated with 2.4 to 4.8 million units of benzathine penicillin G intramuscularly. Neonatal outcomes and signs of congenital syphilis were recorded. RESULTS: Six women had primary, 12 had secondary, and six had early latent syphilis. Sixty-six percent of fetuses (95% confidence interval [CI] 47%, 82%) had either congenital syphilis or detection of Treponema pallidum in amniotic fluid. Sixty-six percent had hepatomegaly, including three fetuses (12.5%, 95% CI 4%, 31%) with ascites. Fetal antitreponemal IgM was detected in three cases. Abnormal liver transaminases were found in 88% (CI 69%, 96%), anemia in 26% (CI 13%, 47%), and thrombocytopenia in 35% (CI 19%, 55%). Maternal treatment was successful in 83% (CI 64%, 93%). Risk of treatment failure was significantly increased when hepatomegaly and ascites were present (P =.01). CONCLUSION: Findings with fetal syphilis are similar to those of neonatal syphilis. We hypothesize that fetal transaminase elevation occurs early in the course of infection; hematologic abnormalities and hydrops occur later. Severity of disease may be associated with risk of treatment failure. PMID- 11384702 TI - Effects of antenatal corticosteroid administration on mortality and long-term morbidity in early preterm, growth-restricted infants. AB - OBJECTIVE: To evaluate the effect of antenatal corticosteroids on mortality, morbidity, and disability or handicap rate in early preterm, growth-restricted infants. METHODS: This case-control study in two tertiary care centers included all live-born singleton infants with growth-restriction due to placental insufficiency, who were delivered by cesarean because of cardiotocographic signs of fetal distress before the beginning of labor at a gestational age of 26-32 weeks during the years 1984-1991. Infants who had been treated antenatally with corticosteroids more than 24 hours and less than 7 days before birth were matched by birth weight, sex, and year of birth with infants whose mothers had been admitted more than 24 hours before delivery but were not treated antenatally with steroids. The main outcome measure was survival without disability or handicap at 2 years corrected age. A sample of 60 case-control pairs would give 81% power to demonstrate 50% increase of this outcome [odds ratio (OR) 3.0] by corticosteroid treatment. Behavior and physical growth were evaluated at school age by questionnaire. RESULTS: The study group and control group consisted of 62 infants each. Survival without disability or handicap at 2 years' corrected age was more frequent in the corticosteroid group [OR 3.2, confidence interval (CI) 1.1, 11.2]. In the long-term follow-up at school age there was a statistically significant negative effect on physical growth (OR 5.1, CI 1.4, 23.8), but no differences in behavior were detected. CONCLUSION: Benefits from antenatal corticosteroids for early preterm, growth-restricted infants appear to outweigh possible adverse effects. PMID- 11384703 TI - Obstetric antecedents to apparent stillbirth (Apgar score zero at 1 minute only). AB - OBJECTIVE: To identify antecedent risk factors for the delivery of an infant with an Apgar score of 0 at 1 minute who is subsequently successfully resuscitated. METHODS: Infants born between January 1986 and February 1999 with 1-minute Apgar score of 0 followed by 5-minute Apgar score above 0 were studied. Each eligible infant was randomly matched with two control infants, born in the same year, with 1-minute Apgar score greater than 0. Hospital records of their mothers were reviewed. The variables were compared between the groups by univariate analysis. Those factors demonstrating significant differences were then analyzed by logistic regression. P <.05 was considered statistically significant. RESULTS: Seventy-four of 81,603 infants (0.9:1000 births) born with an Apgar score of 0 at 1 minute only were compared with 148 control babies. Univariate analysis revealed significant differences between study and control group regarding: gestational age, abruptio placentae, preterm premature rupture of membranes, chorioamnionitis, preeclampsia, small-for-gestational age, male gender, bradycardia, and abnormal fetal heart rate (FHR) other than bradycardia, respectively. Logistic regression of these factors found gestational age, bradycardia, and abnormal FHR to be independent risk factors for the delivery of an apparent stillborn infant. After exclusion of FHR criteria, logistic regression found gestational age (odds ratio [OR] 0.8 per week), male gender (OR 2.5), preeclampsia (OR 3.9), and abruptio placentae (OR 13.6) to be independent risk factors for the delivery of an apparent stillborn infant. CONCLUSION: Preterm birth, male gender, preeclampsia, and abruptio placentae are independently associated with an increased risk of apparent stillbirth. PMID- 11384704 TI - Localization of telomerase hTERT protein and survivin in placenta: relation to placental development and hydatidiform mole. AB - OBJECTIVE: To find if a difference in telomerase or survivin expression exists between non-neoplastic tissues and hydatidiform moles, and explore expression of those proteins in normal placental development, post-term gestation, and preeclampsia. METHODS: Formalin-fixed placental tissues were selected from collections of the Department of Pathology at the University of Colorado. Five specimens of each trimester, five each of preeclamptic and post-term placentas, and 23 molar pregnancies were selected. The telomerase catalytic protein hTERT was localized in placental tissues using the catalyzed signal amplification system, and survivin was localized by conventional immunoperoxidase method. Staining was graded on a scale of zero to 4. RESULTS: hTERT staining was detected in sections of 42 of 48 specimens (23 of 23 hydatidiform moles, 19 of 25 non neoplastic placental tissues). The intensity of staining for hTERT was higher in hydatidiform moles (mean 3.3, median 3) compared with levels in non-neoplastic placental tissues (mean 0.92, median 1) (P <.001). Survivin was detected in 39 of 48 specimens (22 of 23 hydatidiform moles, 17 of 25 non-neoplastic placental tissues). Compared with non-neoplastic tissues (mean 0.88, median 1), survivin levels were elevated in hydatidiform moles (mean 1.35, median 1) (P =.031). CONCLUSION: Survivin and telomerase were increased in hydatidiform moles, suggesting that regulation of apoptosis and stabilization of telomere length might be involved in neoplastic transformation of the placenta. The patterns of expression observed for survivin and telomerase in non-neoplastic placental tissues suggest that the control of apoptosis and stabilization of telomeric DNA might also be involved in normal gestational development. PMID- 11384705 TI - Medically recommended cessation of employment among pregnant women in Georgia. AB - OBJECTIVE: To ascertain the proportion of employed pregnant women who receive medical advice to stop working during pregnancy and to describe their characteristics. METHODS: Data were analyzed from the Georgia Pregnancy Risk Assessment Monitoring System, a surveillance system that surveys new mothers about pregnancy risk factors, health behaviors, and birth-related outcomes. Employment during pregnancy was defined as work for pay for 10 hours or more per week. RESULTS: We studied 1635 women who were employed during pregnancy. A physician or nurse had advised 27.7% (95% CI 24.5%, 30.9%) of them to stop working during pregnancy. Independent predictors of receiving this advice were hospitalization (RR 2.3, 95% CI 1.7, 2.8) and history of previous preterm birth (RR 1.6, 95% CI 1.1, 2.2). Low birth weight (under 2500 g) occurred in 5.8% of women not advised to stop work, in 6.9% of women advised to stop work because of swelling, fatigue, stress, or another reason, and in 13.4% of women advised to stop work because of labor, high blood pressure, or vaginal bleeding (P <.001). Among women advised to stop working in the first through seventh months of pregnancy, 91.7% (95% CI 88.8, 94.5) delivered at 36 or more weeks' gestation. CONCLUSION: Work cessation during pregnancy was commonly recommended in this population and was associated with clinical risk factors and adverse birth outcomes. For some women it resulted in a long period of work absence before delivery. PMID- 11384706 TI - Seasonality in conception of births and influence on late initiation of prenatal care. AB - OBJECTIVE: To determine whether there is a summer peak in conception of births to adolescents (up to 17 years) compared with older teenagers (18-19 years) and adults (20-29 years), and to assess the influence of season of conception on late initiation of prenatal care. METHODS: We analyzed 1,178,607 singleton births to women aged 29 years and younger in Texas between 1994 and 1998. Dates of conception were estimated using last menstrual period and clinical estimates of gestation. Proportions of births conceived per month were assessed for seasonal patterns. Proportions of births with late initiation of prenatal care were also compared by month of conception. The outcomes were further stratified by sociodemographic variables. RESULTS: There was a consistent summer trough (7.5% in August) and year-end peak (9.1% in December) in conception of births to adolescents (P <.001), a pattern similar to that of older teenagers and adults. Among the adolescents, students and non-Hispanic women giving birth presented a secondary early summer peak (8.8% each in May and June) in their conceptions. There was a modest bimodal effect of season of conception on initiation of prenatal care in all age groups. Adolescent conceptions in April-May and September-October were 14-18% and 6% significantly more likely to have late prenatal care compared with other months, respectively. CONCLUSION: Adolescents giving birth in Texas were not more likely to conceive in the summer. They did present seasonal patterns of conception and late initiation of prenatal care similar to older women. PMID- 11384707 TI - In-home nursing care for women with high-risk pregnancies: outcomes and cost. AB - OBJECTIVE: To evaluate a program that provides in-home care to women with pregnancies threatened by preterm delivery (including preterm labor, preterm premature rupture of membranes, and multiple gestation) and women with pregnancy related hypertension. METHODS: Data from hospital discharge summaries were used to compare birth outcomes and cost of care for women in the in-home program and a cohort of women who received in-hospital antenatal care before the new program. Birth outcomes included data for mothers and infants. The sample included 437 women with threatened preterm delivery (n = 228 in-home, n = 209 in-hospital) and 308 with hypertension (n = 155 in-home, n = 153 in-hospital). The cost per woman included all costs of services for mothers and infants. RESULTS: Women at risk of preterm delivery who received in-home care were half as likely to have their infants in the neonatal intensive care unit more than 48 hours (odds ratio 0.53, 95% confidence interval 0.36, 0.78). On average, their infants weighed more (2732 +/- 716 g versus 2330 +/- 749 g, P <.001) and were 2 weeks older at birth (36.1 +/- 3.1 weeks versus 34.0 +/- 4.0 weeks, P <.001). There was a wide range in the total cost per woman and no significant difference between cohorts. For women with hypertension, there were no significant differences between in-home and in hospital cohorts in birth outcomes or costs of care per woman. CONCLUSION: The program with current admission criteria, staffing, and guidelines for antenatal hospital admission provides safe care to women at similar cost to that of hospitalization. PMID- 11384708 TI - Health-related functional status in pregnancy: relationship to depression and social support in a multi-ethnic population. AB - OBJECTIVE: To describe perceived well-being and functional status during uncomplicated late pregnancy among low-income minority women, and to examine the relationship of functional status to depression and social support. METHODS: Hispanic and black women with low-risk pregnancies completed an interview consisting of demographics, the Medical Outcomes Study Short Form 36 (SF-36), Beck Depression Inventory-II (BDI-II), and the Norbeck Social Support Questionnaire. RESULTS: Of the 155 women who were eligible and asked to participate, 41 refused for a participation rate of 74%. Results of the SF-36 showed lowest perceived well-being in the vitality and physical role dimensions. Depressive symptomatology was high, with a mean BDI score of 15 (standard deviation 8.6). Using a BDI score of 14 as the cutoff point, over half of the sample was categorized as having significant depressive symptoms. Significantly lower functional status was seen for depressed subjects in all subscales of the SF-36 compared with nondepressed subjects. Although functional status was negatively correlated with BDI score in all dimensions (r =.23-.69), correlation of SF-36 scores with social support was much weaker (r =.06-.24). CONCLUSION: Elevated levels of depressive symptomatology are strongly correlated with lowered health-related functioning and perceived well-being. Social support is not associated with increased physical or emotional well-being but is weakly associated with mental health as measured by the SF-36. PMID- 11384709 TI - Provider attitudes about gaining consent for perinatal autopsy. AB - OBJECTIVE: To examine the attitudes of neonatologists, obstetricians, midwives, and neonatal nurses toward perinatal autopsy and survey physicians about whom they perceive influence women's decisions on autopsy consent. METHODS: A postal survey that incorporated a questionnaire of eight fictitious case scenarios and combined three factors (confidence of antemortem diagnosis, intention to have future pregnancy, and parental attitude toward autopsy) in various permutations was sent to various Australian physicians and nurses (all consultant neonatologists working in neonatal intensive care units and a sample of consultant obstetricians, midwives, and neonatal nurses in level III maternity hospitals). Respondents were asked to rate how likely they were to seek consent for or suggest autopsies on a seven-point Likert scale (1 = certainly will not, 7 = certainly will). Interactions between factors and respondents were measured by analysis of variance, and differences were compared using Mann-Whitney U, chi(2), and generalized estimating equation tests. RESULTS: The overall response rate was 70% (neonatologists 57%, obstetricians 62%, midwives 77%, and neonatal nurses 75%). Neonatologists (median score 7, interquartile range 7, 7) were more likely to ask for autopsies than neonatal nurses (5; 2, 6) (P <.001), as were obstetricians (7; 7, 7) compared with midwives (6; 3, 7) (P <.001). Physicians rated midwives and neonatal nurses as having some to substantial influence on mothers' decisions about consent for autopsy. CONCLUSION: Physicians are not averse to seeking consent for perinatal autopsies. Midwives and nurses are influenced by the three factors studied, which might negatively influence the consent rate for perinatal autopsies. Intervention strategies aimed at changing nurses' attitudes should be considered. PMID- 11384710 TI - Human immunodeficiency virus type 1-related nucleic acids and papillomavirus DNA in cervicovaginal secretions of immunodeficiency virus-infected women. AB - OBJECTIVE: To evaluate simultaneous human immunodeficiency virus (HIV)-related nucleic acids and human papillomavirus (HPV)-DNA in cervicovaginal secretions of HIV-seropositive women. METHODS: We collected 47 paired blood and cervicovaginal lavage samples from 124 known HIV-1-seropositive women. Proviral HIV-1 DNA, cell associated, and cell-free HIV-1 RNA in cervicovaginal secretions were quantitatively evaluated by competitive polymerase chain reaction (PCR) and reverse transcription PCR. Polymerase chain reaction and subsequent restriction fragment length polymorphism analysis of PCR products were used to detect HPV types 6, 11, 16, 18, 31, 33, 35, and 56. RESULTS: Proviral HIV-1 DNA, cell associated, and cell-free HIV-1 RNA were detected in 52.4% (65 of 124), 38.7% (48 of 124), and 33.9% (42 of 124) of lavage samples, respectively. Human papillomavirus-DNA in cervicovaginal secretions was detected in 64% (79 of 124) of participants. The rate of detection of HPV types of intermediate to high oncogenic risk was higher in HIV-positive women who tested positive for cell associated (odds ratio [OR] 3.57, 95% confidence interval [CI] 1.17, 11.12) or cell-free (OR 4.63, 95% CI 1.42, 15.51) HIV-1 RNA in cervicovaginal secretions than their counterparts who tested negative. Logistic regression analysis showed that the association between HPV infection and the detection of HIV-1 RNA in cervicovaginal secretions persisted after adjustment for potential confounders such as CD4+ cell counts, HIV-1 RNA in plasma, use of antiretroviral drugs, vaginal infection, and regular condom use. In univariable and multivariable analysis, HPV-DNA detection was associated with amounts of cell-free and cell associated HIV-1 RNA in cervicovaginal secretions (chi(2) for trend 10.35, and 9.84, P =.001 and.002, respectively). CONCLUSIONS: The rate of HPV detection in the genital tract of HIV-1-seropositive women is associated with the amount of cell-associated and cell-free HIV-1 RNA present in cervicovaginal secretions. The association does not appear to be attributable entirely to the effect of HIV related immunodepression. PMID- 11384711 TI - Adieu. PMID- 11384713 TI - Impact of infectious diseases on women's health: 1776-2026. AB - In this commentary, the author reviews the impact of infectious diseases on women's health, beginning with the late 18th century when clinically recognized infections took the heaviest toll on women. The review continues through the 19th and 20th centuries when public health measures and the advent of antibiotic agents led to apparent control of infection, and ends with a forecast for the next 25 years. Foreboding considerations for the future include spread of human immune deficiency virus infection and other sexually transmitted diseases and the development of new infections, antibiotic-resistant bacteria, and the recognition of infection as a cause of many obstetric-gynecologic and chronic disorders. Obstetricians and gynecologists can contribute to the solution of these problems individually and collectively. PMID- 11384712 TI - Long hours and little sleep: work schedules of residents in obstetrics and gynecology. AB - OBJECTIVE: To investigate residents' work schedules and their attitudes toward limiting their hours. METHODS: An anonymous survey regarding resident work hours and call schedules was administered to the 4674 obstetric-gynecologic residents who took the year 2000 Council on Resident Education in Obstetrics and Gynecology in-training examination. RESULTS: A total of 4510 surveys were analyzed (96.5%). Three of four (75.5%) respondents reported working between 61 and 100 hours each week. Most (71.3%) reported sleeping less than 3 hours while on night call. Eight of ten reported having postcall clinical responsibilities. The reported number of hours on call declined and the reported number of hours of sleep increased with year of residency. Three of four residents wanted limits on their work hours. Residents who reported longer on-call hours or less sleep during night shift were significantly more likely to want a restriction on work hours. Fatigue was the most commonly selected reason (77.6%) followed by "need more personal time" (76.3%), and "fear of compromising quality of care" (59.8%). Women were more concerned about fatigue than were men. Among residents who did not want work hour restrictions, "additional surgical experience" was the most commonly selected reason (69.0%). CONCLUSION: Residents in obstetrics and gynecology report working long hours, and experiencing periods of little sleep. Most want their work hours to be limited. Fatigue is a major concern among residents that want their hours limited. A sizable minority worries that such limits might also limit their experience. PMID- 11384714 TI - Potassium channels in T lymphocytes: toxins to therapeutic immunosuppressants. PMID- 11384715 TI - Divergent Pseudomonas exotoxin A sensitivity in normal and transformed liver cells is correlated with low-density lipoprotein receptor-related protein expression. AB - Pseudomonas exotoxin A (PEA) is an extracellular virulence factor produced by the opportunistic human pathogen Pseudomonas aerguinosa. PEA intoxification begins when PEA binds to the low-density lipoprotein receptor-related protein (LRP). The liver is the primary target of systemic PEA, due largely to the high levels of functional LRP expressed by liver cells. Using a 3H-leucine incorporation assay to measure inhibition of protein synthesis we have demonstrated that normal (BNL CL.2) and transformed (BNL 1ME A7R.1) liver cells exhibit divergent PEA sensitivity; with BNL 1ME A7R.1 cells demonstrating greater PEA sensitivity than their non-transformed counterparts. The receptor-associated protein, a LRP antagonist, decreased PEA toxicity in BNL 1ME A7R.1 cells, confirming the importance of the LRP in PEA intoxification in this cell type. Increased PEA sensitivity in BNL 1ME A7R.1 cells was associated with increased functional cell surface LRP expression, as measured by alpha2-macroglobulin binding and internalization studies, and increased LRP mRNA levels, as determined by Northern blot analysis. Interestingly, BNL CL.2 cells were more sensitive than BNL 1ME A7R.1 cells to conjugate and mutant PEA toxins that do not utilize the LRP for cellular entry. These data demonstrate that increased LRP expression is an important mechanism by which PEA sensitivity is increased in BNL 1ME A7R.1 transformed liver cells. PMID- 11384716 TI - Precursor nucleotide sequence and genomic organization of BmTXKS1, a new scorpion toxin-like peptide from Buthus martensii Karsch. AB - Scorpion venom contains a variety of small peptides, which can modulate Na+, K+, Ca2+ and Cl- channel conductance in excitable and non-excitable tissues. A novel full-length cDNA encoding a new toxin-like peptide (named BmTXKS1) was isolated from the venom gland cDNA library of Buthus martensii Karsch. The precursor consists of 60 amino acid residues, with a putative signal peptide of 28 residues and an extra residue, and a mature peptide of 31 residues with an amidated C terminal. BmTXKS1 shared close homology with BmP01 in 5'UTR and the region encoding the putative signal peptide; especially, the positions of six cysteines are highly conserved among BmTXKS1, PbTX1 and P01-type subfamily of scorpion K+ channel toxins, suggesting that they all should present a common three dimensional fold, namely the Cysteine-Stabilized alphabeta(CSalphabeta) motif. By PCR amplification of the genomic region encoding BmTXKS1, we have confirmed the identity of our cloned cDNA, and found that BmTXKS1 gene contains an intron, which is completely identical with that of the characterized scorpion K+-channel ligands in the size, consensus junctions, putative branch point and A+T abundance. PMID- 11384717 TI - Electrophysiological and hemolytic activity elicited by the venom of the jellyfish Cassiopea xamachana. AB - In this study, we determined hemolysis activity in human and sheep erythrocytes, and characterized the electrical responses in Xenopus oocyte membrane elicited by the venom of the jellyfish Cassiopea xamachana (Cx). The Cx venom produced hemolysis in both species, being more potent on human red cells. The electrophysiological study showed that the Cx venom elicited three different responses in the oocytes. One current was generated in all the oocytes tested and corresponded with a slow inward current (I(Cx)) associated with an increase in membrane conductance. I(Cx) was concentration-dependent and had a reversal potential of -10.3+/-0.4 mV. Ionic substitution studies indicated that the conductive pathway was mainly permeable to cations and non-selective. The oocyte membrane resistance was completely recovered after washout of the venom, this suggested that the effect was due to generation of a specific membrane conductance as opposed to a possible non-specific membrane breakdown. A comparative study with three distinct native cationic channels present in the oocyte membrane [i.e. (1) hemi-gap-junction channels, (2) mechanosensitive channels, and (3) the ouabain-sensitive channel activated by palytoxin], showed that I(Cx) might correspond to opening of mechanosensitive channels or to activation of an unknown cationic channel located in the oocyte membrane. The bioactive fraction eliciting I(Cx) were peptides and was separated from two other peptidic hemolytic fractions by chromatography. PMID- 11384718 TI - Correlation of cleavage of SNAP-25 with muscle function in a rat model of Botulinum neurotoxin type A induced paralysis. AB - Injection of botulinum neurotoxin serotype A (BoNT/A) into muscle results in cleavage of the synaptosomal associated protein of 25 kDa (SNAP-25) and relatively long-term paralysis. However, nerve-terminal sprouting, which appears to require intact SNAP-25, has been reported to occur much earlier. The difference between the long-term paralysis induced by injection of BoNT/A and the short time needed for sprouting led us to investigate the relationship between BoNT/A catalyzed cleavage of SNAP-25 and muscle function. The effect of BoNT/A on SNAP-25 present in nerve endings innervating gastrocnemius muscles of rats was monitored over time. Cleaved SNAP-25 was found in nerve terminals innervating the muscles within 24h of inoculation with BoNT/A and was present more than 2 months later. Comparison of the ratios of cleaved to intact SNAP-25 from the onset of BoNT/A-induced paralysis until function was regained indicated that paralysis was probable when the ratio of cleaved to intact SNAP-25 was greater than 0.35. PMID- 11384719 TI - Effects of antivenom on Buthus occitanus tunetanus (Bot) scorpion venom pharmacokinetics: towards an optimization of antivenom immunotherapy in a rabbit model. AB - The pharmacokinetic parameters of Bot venom were determined in a rabbit model using a specific sandwich type ELISA. After intravenous injection, Bot venom seems to follow a three-compartment pharmacokinetic open model. However, after subcutaneous injection, the distribution and elimination kinetics of Bot venom are best characterized by a bi-compartment pharmacokinetic open model. Bot venom is completely absorbed from its SC injection site, since the absolute bioavailability is higher than 95%; the maximum plasma venom concentration is reached between 30 and 60 min after venom injection. Bot venom diffuses rapidly to tissues and is distributed in a high body volume. The total body clearance of Bot venom is relatively high in agreement with a low mean residence time. Antivenom immunotherapy experiments were carried out in the rabbit model, in order to select the most appropriate strategy for the adequate use of this treatment. The effects of the route, the dose and the delay of antivenom injection on Bot venom pharmacokinetic parameters and on the antivenom immunotherapy efficacy were then studied. These studies indicated in particular that: (1) the injection of a minimal neutralizing antivenom dose is required for a complete and permanent neutralization of circulating venom antigens; this dose is named minimal (threshold) efficacious antivenom dose; (2) the intramuscular route is not the most appropriate way for antivenom injection; and (3) a delayed antivenom immunotherapy remains efficacious especially on the neutralization of the remaining circulating venom. In short, these experimental studies show that early intravenous injection of an appropriate antivenom dose (at least the threshold efficacious dose) is the indicated way for a rapid and permanent neutralization of circulating scorpion venom toxins. PMID- 11384720 TI - Hemorrhagic activity and muscle damaging effect of Pseudomonas aeruginosa metalloproteinase (elastase). AB - Pseudomonas aeruginosa elastase, a Bacillus subtilis thermolysin-like zinc proteinase was examined for hemorrhagic activity and its effect on muscle and endothelial cells. Subcutaneous and intramuscular injections of elastase into mice caused severe hemorrhage with an acute increase of creatine phosphokinase activity in serum. The elastase also possessed fibrinogenolytic and fibrinolytic activities. The Aalpha and Bbeta chains of fibrinogen were completely hydrolyzed as demonstrated by their electrophoretic disappearance on SDS polyacrylamide gels. The pathological study indicates that elastase induces changes in the structure of the vascular wall and causes leakage of the plasma component and red and white blood cells into the extravascular tissue. This is further supported by results showing injury to cultured endothelial cells and macrophages. These data indicate that P. aeruginosa elastase directly affects endothelial cells and destroys the basement membrane of blood vessels to cause hemorrhage. Since fibrinogenolytic activity is an additional component of this elastase and this activity induces the hemorrhagic tendency, the damage in tissues could become increasingly severe. PMID- 11384721 TI - Neutralization of dermonecrotic and lethal activities and differences among 32-35 kDa toxins of medically important Loxosceles spider venoms in Brazil revealed by monoclonal antibodies. AB - Neutralization of dermonecrotic and lethal activities and differences among the principal toxic proteins (32-35 kDa) of medically important Loxosceles spider venoms in Brazil (Loxosceles gaucho, Loxosceles laeta and Loxosceles intermedia) were studied using monoclonal antibodies (MAbs) produced against the dermonecrotic component (35 kDa) of L. gaucho venom. MAb titers were 512,000 to homologous venom, between 2000 and 64,000 for L. intermedia venom and between 1000 and 64,000 for L. laeta venom. By Western blotting, MAbs could recognize mainly the 35 kDa protein of L. gaucho venom and with less intensity the 35 kDa protein of L. intermedia venom. These MAbs also recognized weakly or did not recognize the 32 kDa component of L. laeta venom. Only MoALg1 showed high affinity for L. gaucho venom and neutralized in vivo 90-97% of the dermonecrotic activity, besides delaying the lethality induced by homologous venom. MoALg1 maintained its capacity to neutralize the dermonecrotic activity, even when administered (i.v.) 6h after envenoming (i.d.). All MAbs obtained failed to neutralize the toxic activities of the heterologous venoms.These results suggest that different epitopes are present in the protein responsible for the dermonecrotic activity of Loxosceles venoms, and confirm the participation of other venom components during the local reaction process. This study also confirms the importance of antibodies for neutralization of dermonecrotic activity, even when administered some hours after envenoming, and emphasizes the differences of composition and toxicity of medically important Loxosceles venoms. These findings must be considered in order to improve loxoscelism immunotherapy. PMID- 11384722 TI - Structures involved in production, secretion and injection of the venom produced by the caterpillar Lonomia obliqua (Lepidoptera, Saturniidae). AB - The number of accidents caused by injection of the venom of Lonomia obliqua caterpillars in Southern Brazil has increased in the last years. Even though this kind of envenomation has an important social and medical impact, nothing is known about the cellular structures responsible for the production and secretion of this venom. Here we identify and analyse morphological structures possibly responsible for the production and secretion of the active principles of the venom, as well as the histological relationship of these structures with the urticating spines of L. obliqua. Detailed microscopic observations showed that: (a) L. obliqua has a complex tegument, with several cuticular specializations, (b) there are no pores along the tegument neither in the spines and (c) the spines bear a hollow canal--where the venom is deposited--and an area that can be easily broken when touched, releasing the venom. Histological and histochemical techniques revealed that: (a) there is no single gland cell that produces the venom, (b) a secretory epithelium, composed of cells containing vesicles that increase in size and number as they reach the apical region, underlies the tegument and the spines and is responsible for secretion of the venomous substances and (c) the venom is deposited in the subcuticular space and at the tips of the spines. PMID- 11384723 TI - Decreased parasympathetic activities in Malayan krait (Bungarus candidus) envenoming. AB - Three patients were bitten by the Malayan krait (Bungarus candidus). The patients developed ptosis and generalized muscle weakness which later progressed to respiratory paralysis. All patients showed evidence of decreased parasympathetic activity manifested by mydriasis, hypertension and tachycardia. No specific antivenom was available. All patients received assisted ventilation and supportive treatment. The other forms of treatment included administration of neostigmine, the banded krait (Bungarus fasciatus) antivenom (Thai Red Cross) and plasmapheresis without beneficial response. Two patients recovered. The other patient had permanent brain damage due to anoxia from two episodes of cardiac arrest. While hypertension resolved 6-60 days after admission, mydriasis and tachycardia persisted after discharge in all patients for between 7 days and 2 years. One patient had constipation and defect in micturition which still persisted 2 years after the bite. Decreased parasympathetic activities in Malayan krait bite are perhaps not uncommon and should be examined. PMID- 11384724 TI - Binding of anticoagulation factor II from the venom of Agkistrodon acutus with activated coagulation factor X. AB - Anticoagulation factor II (ACF II) from the venom of Agkistrodon acutus has been identified as a binding protein to activated bovine coagulation factor X (FXa) by the method of polyacrylamide gel electrophoresis and high performance liquid chromatography. This protein formed a 1:1 complex with FXa in the presence of Ca2+ ions, and the maximal binding of ACF II to FXa occurred at the concentration of Ca2+ ions of about 1mM. The binding of Ca2+ ions to ACF II was analyzed by equilibrium dialysis and two Ca2+-binding sites with different affinities were identified. At pH 8.0, the apparent association constant K1 and K2 values for these sites were (1.1+/-0.3)x10(5) M(-1) and (1.7+/-0.4)x10(4) M(-1) (mean+/-SE, n=4), respectively. It was evident from the observation of Ca2+-induced changes in the intrinsic fluorescence of ACF II that ACF II underwent a conformational change upon binding of Ca2+ ions. The occupation of both Ca2+-binding sites in ACF II requires a concentration of Ca2+ ions of about 1mM, which is equal to the effective concentration of Ca2+ ions required for maximal binding of ACF II to FXa, and for the maximal Ca2+-induced enhancement of emission fluorescence of ACF II. It can be deduced from these results that the occupation of both Ca2+-binding sites in ACF II with Ca2+ ions and subsequent conformational rearrangement should be essential for its recognition of Ca2+-mediated conformational changes of FXa. PMID- 11384725 TI - Acute and delayed thermoregulatory response of mice exposed to brevetoxin. AB - Thermal dysthesia, characterized by a painful sensation of warm and cool surfaces, is one of many ailments in humans exposed to various marine algal toxins such as brevetoxin (PbTx). There is no animal model to study thermal dysthesia and little is known of the mechanism of action. There is also little known on the acute and delayed thermoregulatory effects of PbTx. In this study, we developed a behavioral system to assess the possible development of thermal dysthesia in mice exposed to PbTx. Female mice were implanted with radiotransmitters to monitor core temperature (Tc) and motor activity (MA). In one experiment, mice were dosed with the control vehicle or 180 microg/kg PbTx and placed on a floor temperature gradient to measure the selected foot temperature (SFT) while air temperature was kept constant. PbTx-treated mice underwent a 10 degrees C reduction in SFT concomitant with a 3 degrees C reduction in Tc within 30 min after exposure. In another study, Tc and MA were monitored in mice maintained in their home cages after dosing with 180 microg/kg PbTx. Tc but not MA increased for 2-5 days after exposure. SFT was unaffected by PbTx when tested 1-12 days after exposure. However, PbTx-treated mice underwent an increase in Tc when placed in the temperature gradient for up to 12 days after exposure. This suggests that PbTx augments the stress-induced hyperthermia from being placed in a novel environment. Overall, acute PbTx exposure leads to a regulated reduction in Tc as characterized by a preference for cooler SFTs and a reduced Tc. Thermal dysthesia was not apparent, but the exaggerated hyperthermic response with a normal SFT in the temperature gradient may suggest an altered processing of thermal stimuli in mice treated with PbTx. PMID- 11384726 TI - Isolation, characterization and quantification of microcystins (heptapeptides hepatotoxins) in Microcystis aeruginosa dominated bloom of Lalla Takerkoust lake reservoir (Morocco). AB - This paper presents the first data on the identification, characterization and quantification of microcystins isolated from both an extract of a cyanobacteria natural bloom, collected from a eutrophic Moroccan reservoir (Lalla Takerkoust, Marrakesh) and an isolated strain cultivated under laboratory conditions. The isolation and purification of toxins was performed by reverse phase HPLC and then characterized by amino acid analysis and fast atom bombardment mass spectrometry (FAB-MS). Chemical characterization of the toxins from the bloom revealed variants of microcystins such as Mcyst-LR, Mcyst-RR, Mcyst-YR and [D-Asp3]Mcyst LR. However, the Microcystis aeruginosa strain produced only Mcyst-RR. Using an ELISA assay the total microcystin contents of eight bloom samples collected from 1994 to 1997 ranged from 0.7 to 8.8 microg/mg of lyophilized material. The two isolated Microcystis strains contained higher amounts of microcystins (0.65 microg/ mg of dry weight) than the Pseudanabaena strains (0.021 microg/mg of dry weight). Our results show that the presence of cyanobacteria toxins in water used for drinking in a North African country may be regarded as an health hazard. These results are a contribution to the knowledge of the biogeography of toxic cyanobacteria and their toxins, namely in north African countries. PMID- 11384727 TI - Involvement of nitric oxide on the peritoneal neutrophil influx induced by staphylococcal enterotoxin B in mouse. AB - In this study, the role of nitric oxide (NO) on neutrophil migration induced by staphylococcal enterotoxin B (SEB) in the mouse peritoneal cavity was investigated. The NO synthase inhibitors L-NAME and aminoguanidine, as well as dexamethasone, markedly reduced SEB-induced neutrophil influx. In mice with an increased population of peritoneal macrophages, the inhibition of SEB-induced neutrophil influx by these agents was significantly lower. The in vivo treatment with aminoguanidine inhibited only the iNOS activity, whereas L-NAME inhibited both the cNOS and iNOS activities. In conclusion, NO modulates the neutrophil migration in response to SEB through the activity of an iNOS isoform. PMID- 11384728 TI - The validity of two HPLC methods and a colorimetric PP2A assay related to the mouse bioassay in quantification of diarrhetic toxins in blue mussels (Mytilus edulis). AB - Validity of two HPLC methods and a PP2A assay in relation to the mouse bioassay for diarrhetic shellfish poisoning (DSP) toxins was evaluated. The mouse bioassay for DSP toxins was performed on a total of 177 mussel samples from the Sognefjord, Norway, using diethyl ether in the final step of extraction. For fluorimetric HPLC analyses, either 4-bromomethyl-7-methoxycoumarin (BrMMC) or 9 anthryl diazomethane (ADAM) were used for analysis of 48 and 118 of the samples, respectively. The colorimetric PP2A inhibition assay was performed on all 177 samples that were analysed with the mouse bioassay. When comparing the HPLC BrMMC, the HPLC-ADAM and the PP2A assays with the mouse bioassay, cut off values of < or =4, 5 and 6 microg okadaic acid (OA) equivalents (eq.)/5 g digestive gland (DG) was used. With reference to the results from the mouse bioassay, the total number of failure and correct classification by HPLC-ADAM and the PP2A method was compared for the three cut off values. No significant differences between the methods were detected. However, all differences were found in favour of HPLC-ADAM. All three methods could replace the mouse bioassay in detecting levels of diarrhetic toxins approved internationally for safe consumption of mussels. However, HPLC-ADAM seems to be the method of choice. PMID- 11384729 TI - Repeatability and validity of a fluorimetric HPLC method in the quantification of yessotoxin in blue mussels (Mytilus edulis) related to the mouse bioassay. AB - Repeatability and validity of a fluorimetric HPLC method in quantification of yessotoxin (YTX) in mussels related to the mouse bioassay was studied. Blue mussels (Mytilus edulis) from the Sognefjord, Norway were sampled from March to November, 1997, and October to December, 1998. A total of 75 samples were analysed for YTX by HPLC using 4-[2-(6,7-dimethoxy-4-methyl-3-oxo-3, 4 dihydroquinoxalinyl) ethyl]-1,2,4-triazoline-3, 5-dione (DMEQ-TAD) as a fluorimetric derivatization agent. Among these, 28 of the samples were analysed by HPLC in duplicate. All samples were analysed by the mouse bioassay using both chloroform and ether in the final step of extraction. The duplicate measurements using HPLC was found equal and the method repeatable (p<0.05). The absolute difference between the two measurements was found to increase with increasing level of measurements. This significant positive correlation (p<0.05) was mainly due to concentrations of YTX higher than 200 microg/100g mussel meat. However the precision of the results obtained was not found to be less in the upper level than in the lower level. Based on the internal correlation analysis including the mouse bioassay and the HPLC method a cut-off value of < or =10 microg YTX/5 g digestive gland was found preferable. The mouse bioassay of ether extracts often failed to detect high levels of YTX, and as demonstrated by the low kappa-values, the agreement between the mouse bioassay of ether extracts and the HPLC method was very weak. The HPLC method was found to give repeatable results and thereby found to be reliable. Consequently, the HPLC method seems to the method of choice for detection and quantification of YTX in mussels when compared with the mouse bioassay. PMID- 11384731 TI - Investigations into the inhibitory effects of microcystins on plant growth, and the toxicity of plant tissues following exposure. AB - The cyanobacterial toxins microcystins are known to affect a number of processes in plant tissues, and their presence in water used for irrigation may have considerable impact on the growth and development of crop plants. In this study, two plant bioassays were employed to investigate the phytotoxic effects of microcystins. A plant tissue culture assay revealed that the growth and chlorophyll content of Solanum tuberosum L. cultures was inhibited at microcystin LR concentrations of 0.005 and 0.05 microg x cm(-3), respectively. A previously developed bioassay was also employed to determine the effects of three commonly occurring microcystin variants on the growth of Synapis alba L. seedlings. Microcystins-LR, -RR, and -LF inhibited the growth of seedlings, with GI50 values of 1.9, 1.6 and 7.7 microg x ml(-1), respectively. The growth of Phaseolus vulgaris was also examined in the presence of microcystin-LR. The toxin was found to have little effect on growth for up to 18 days, but impaired the development of the roots of exposed plants, causing them to take up approximately 30% less growth medium than those grown in the absence of toxin. Microcystin was also detected in the tissues of exposed plants using a commercially available ELISA kit, suggesting that the uptake of these toxins by edible plants may have significant implications for human health. PMID- 11384730 TI - Tolerance to the antinociceptive effect of Crotalus durissus terrificus snake venom in mice is mediated by pharmacodynamic mechanisms. AB - Crotalus durissus terrificus venom exerts central and peripheral antinociceptive effect mediated by opioid receptors. The present work investigated the tolerance to the antinociceptive effect of the venom and characterised the mechanisms involved in this phenomenon. The hot plate test, applied in mice, was used for pain threshold determination. The venom (200 microg/kg) was administered by oral route, daily, for 14 days, and the nociceptive test was applied before and on days 1, 7 and 14 of the treatment. Prolonged treatment with venom lead to the development of tolerance to the antinociceptive effect. Tolerant animals exhibited increased sodium pentobarbital-induced sleeping time, although total hepatic microsomal cytochrome P450 was not altered. The antinociceptive effect of a single dose of venom (200 microg/kg) is mediated by kappa opioid receptors. Mice long-term-treated with venom showed cross-tolerance to U-TRANS, an agonist of kappa-opioid receptor, but not to morphine or DAMGO, two mu-opioid receptor agonists. Prolonged administration of venom did not cause symptoms of abstinence syndrome. These data indicate that prolonged treatment with C. durissus terrificus venom induces tolerance to the antinociceptive effect and that pharmacodynamic mechanisms are involved in the genesis of this phenomenon. PMID- 11384732 TI - Cardiovascular effects of equinatoxin III from the sea anemone Actinia equina (L.). AB - Equinatoxin III is the most hemolytic, and the least lethal of the three basic proteins isolated from the sea anemone Actinia equina (L.). Its LD50 in mice is 83 microg/kg. Preliminary results on Wistar rats have suggested cardiorespiratory arrest as a putative cause of death, but the mechanism of its action has not yet been studied. So far only equinatoxin II has been investigated more thoroughly. As equinatoxin II is less lythic, but more toxic, than equinatoxin III (its LD50 in mice=35 microg/kg), it may be assumed that haemolysis with a consequent rise in plasma potassium level is not the major factor in the lethality of equinatoxins. To assess the relative contribution of hyperkalemia in the lethality of the toxin in rat, the effects of equinatoxin III were compared to the effects of hyperkalemia caused by the injection of KCl giving the same final concentration of K+ in the plasma as that observed after an i.v. injection of 3LD50 of equinatoxin III. As coronary vasoconstriction may be an important mechanism of the cardiotoxic action of equinatoxins, the effect of EqT III on isolated porcine coronary arteries was studied by measurements of smooth muscle tension in the presence of 1-100 nM equinatoxin III. The results revealed that animals survive the elevated K+ plasma concentration caused by an i.v. application of KCl. This suggests that equinatoxin III induced haemolysis is not the major mechanism of equinatoxin III lethality. However, equinatoxin III increases the potassium induced contractions of coronary smooth muscle for 289+/ 29%, suggesting that coronary vasoconstriction may be an important factor in the cardiotoxic effects of equinatoxin III. PMID- 11384733 TI - Novel in vitro assays for assessing the haemorrhagic activity of snake venoms and for demonstration of venom metalloproteinase inhibitors. AB - Standard methods used for assessing the haemorrhagic toxicity of snake venoms and the effectiveness of antivenoms are laborious, expensive and involve the use of large numbers of laboratory animals. This paper examined the feasibility of using a gelatin degradation ELISA for preliminary screening of snake venom metalloproteinases (MPs). Potent gelatinolytic activity was observed in venoms from snakes of the family Viperidae and, as expected, little or no activity was evident in the venoms of snakes that induce neurotoxic pathology (most elapids). A reverse gelatin zymography assay was used on a variety of venoms to demonstrate a number of inhibitors of MP activity, the first such demonstration of its kind. PMID- 11384734 TI - Structure-activity relationships for human estrogenic activity in zearalenone mycotoxins. AB - Zearalenones are estrogenic Fusarium mycotoxins consisting of a resorcinol moiety fused to a 14-member macrocyclic lactone. Using an improved MCF7 human breast cell proliferation assay, we have compared the estrogenicity of 17 chromatographically-homogeneous zearalenones. Both similarities and substantial differences from published results in intact animal systems were observed. Substantial human estrogenicity was retained even in analogs lacking hydroxylation on the aromatic and macrocyclic rings. PMID- 11384735 TI - Effect of Laticauda semifasciata (sea snake) venom on chloride transport across the frog skin. AB - Addition of Laticauda semifasciata venom to the outside bathing solution of isolated frog skin elicited sustained increases in transepithelial potential difference, short-circuit current and transepithelial electrical resistance. The Laticauda semifasciata venom-induced short-circuit is carried by a net active Na+ transfer from the outside to the inside bathing solution while the increase in transepithelial resistance is accounted for by a decrease in the outside to inside Cl- flux. PMID- 11384736 TI - Genetic alterations at the nuclear localization signal of the RB2/p130 gene occur in lymphoid tumor but not in osteosarcoma cell lines. PMID- 11384737 TI - An expanded view of inositol signaling. PMID- 11384738 TI - Regulation of cardiac myocyte protein synthesis through phosphatidylinositol 3' kinase and protein kinase B. PMID- 11384739 TI - Spontaneous occurrence of an inhibitor of protein kinase C localization in a thyroid cancer cell line: role in thyroid tumorigenesis. PMID- 11384741 TI - The protein kinase activity modulation sites: mechanisms for cellular regulation targets for therapeutic intervention. PMID- 11384740 TI - Regulation and role of the atypical PKC isoforms in cell survival during tumor transformation. PMID- 11384742 TI - Importance of dihydropyrimidine dehydrogenase (DPD) deficiency in patients exhibiting toxicity following treatment with 5-fluorouracil. PMID- 11384743 TI - Regulation of retinoic acid metabolism. PMID- 11384744 TI - Comparison of the properties of human breast cancer cells: MCF-7 and MCF-7 cells selected for resistance to etoposide. PMID- 11384745 TI - The enhanced permeability and retention (EPR) effect in tumor vasculature: the key role of tumor-selective macromolecular drug targeting. PMID- 11384746 TI - Adipocyte expression of a neuronal RNA-binding protein. PMID- 11384747 TI - Apoptosis signaling pathways mediated by cyclooxygenase-2 inhibitors in prostate cancer cells. PMID- 11384748 TI - Novel procaspase-3 activating cascade mediated by lysoapoptases and its biological significances in apoptosis. PMID- 11384749 TI - Effect of malignant transformation on lactate levels of human mammary epithelial cells. PMID- 11384750 TI - The Ras-Raf relationship: an unfinished puzzle. PMID- 11384751 TI - Control of pyruvate dehydrogenase kinase gene expression. PMID- 11384753 TI - Pathogenesis of adult-onset type II citrullinemia caused by deficiency of citrin, a mitochondrial solute carrier protein: tissue and subcellular localization of citrin. PMID- 11384754 TI - Human heparanase: a molecular determinant of brain metastasis. PMID- 11384752 TI - Synergistic effects of pi3k/akt on abrogation of cytokine-dependency induced by oncogenic raf. PMID- 11384755 TI - Nuclear inositol lipid signaling. PMID- 11384757 TI - A critical survey of the structure-function of the antisense oligo/RNA heteroduplex as substrate for RNase H. AB - The aim of this review is to draw a correlation between the structure of the DNA/RNA hybrid and its properties as a substrate for the RNase H, as well as to point the crucial structural requirements for the modified AONs to preserve their RNase H potency. The review is divided into the following parts: (1) mechanistic considerations, (2) target RNA folding-AON folding-RNase H assistance in AON/RNA hybrid formation, (3) carbohydrate modifications, (4) backbone modifications, (5) base modifications, (6) conjugated AONs, (7) importance of the tethered chromophore in AON for the AON/RNA hybrid interactions with the RNase H. The structural changes in the AON/RNA hybrid duplexes brought by different modifications of the sugar, backbone or base in the antisense strand, and the effect of these changes on the RNase H recognition of the modified substrates have been addressed. Only those AON modifications and the corresponding AON/RNA hybrids, which have been structurally characterized by spectroscopic means and functionally analyzed by their ability to elicit RNase H potency in comparison with the native counterpart have been presented here. PMID- 11384758 TI - Fibrous stationary phase in capillary electrochromatography. AB - Capillary electrochromatography (CEC) using fibrous cellulose acetate (CA) stationary phase was investigated. The advantage of this fiber-packed column is relatively easy preparation process compared with other conventional CEC columns, such as particle-packed and wall-coated capillaries. CA fibers are manually packed into a capillary with two guide liners and fixed with a frit at the column inlet. The separation characteristics of this column were investigated using n alkyl p-hydroxybenzoates (parabens) as the sample probe. It has been demonstrated that the use of a short column length and a specially designed tee-connector as the injection device should make the separation performance and efficiency much higher on the fiber-packed columns. Sufficient separation between methyl and n butylparabens is obtained on the 5-cm-packed column and linear relationships between the injection time and the peak area are observed. Bubble formation is not encountered during the analysis. PMID- 11384759 TI - A systematic approach to the complete study of a signaling molecule: ribosomal p90rsk as an example. AB - Ribosomal p90rsk is a kinase of central importance in transducing mitogenic signals from an activated receptor to the cell nucleus and for protein synthesis. Here, we analyze the optimal steps to fully describe this kinase in both normal neutrophils and leukemic cell lines. These are: (i) immunological analyses (immunoblotting and immunoprecipitation); (ii) enzyme activity assays (in vitro and "in-gel"); and (iii) immunobiochemical combination methods (immunoprecipitation/kinase assay, immunoprecipitation/"in-gel" assay and ion exchange chromatography/immunoblotting). For the enzyme assays, we describe a novel method to measure ribosomal p90rsk kinase activity "in-gel", based on a renatured-protein method that allows for the direct quantitation of enzyme activity. Finally, we present an algorithm that can be readily implemented to the quantification of the extent of stimulation of a kinase in response to a particular extracellular stimuli. In our case, it was found that activation of p90rsk was higher in proliferating leukemic cells than in mature neutrophils, indicating that a suppression of key signal transduction links could contribute to the maturational arrest typical of acute leukemia. All the techniques and strategies described here for p90rsk could be easily extrapolated to the study of any signal transduction molecule, provided it has a phosphotransferase activity. PMID- 11384760 TI - Towards time-resolved, coupled structure-function information on carotenoid excited state processes: X-ray and optical short-pulse double resonance spectroscopy. AB - A novel soft X-ray and optical short-pulse double resonance spectroscopic technique tailor-made to elucidate processes involving the optically forbidden S1 (2(1)A(g)) state of carotenoids in native biological samples (e.g., photosynthetic antenna complexes) is described. The principle relies on probing the near carbon K-edge absorption of the optically excited sample with soft X rays generated by a laser-induced plasma. A first application concerns location of the 2(1)A(g) state of beta-carotene in vitro. Further applications are proposed. PMID- 11384761 TI - Stereoselective synthesis of (S)-(+)-Naproxen catalyzed by carboxyl esterase in a multicompartment electrolyzer. AB - The stereoselective hydrolysis of racemic 2-substituted propionates, catalyzed by carboxyl esterase, provides a cost-competitive route to produce the optically pure, anti-inflammatory drug Naproxen. In the present work, we describe the application of the multicompartment electrolyzer reactor (ME) for the stereoselective hydrolysis of a racemic Naproxen ester, (R,S)-ethoxyethyl-[2-(6 methoxy-2-naphtyl)]propionate, catalyzed by a carboxyl esterase. The enzyme was trapped in a reactor chamber, delimited by two isoelectric membranes encompassing the pI value of the enzyme, together with the neutral substrate. After 90 min, a conversion of 45% was obtained with an enantiomeric excess of 84%. The reaction product, (S)-(+)-Naproxen, was electrophoretically removed in continuous from the reaction chamber and collected in a contiguous, more acidic chamber, separated from the enzyme and from the unreacted substrate. Moreover, at the end of the reaction, it was possible to recover the enzyme from the reactor and use it again. PMID- 11384762 TI - Development of a new method of analysing chemotactic deactivation of human neutrophil granulocytes. AB - Measurement of chemotactic migration of human neutrophil granulocytes (PMN) induced by chemotaxins serves as a simple and reliable method for assessing the expression of chemotaxin receptors. Incubation of PMN with a certain chemotaxin leads to a diminished chemotactic migration towards this chemotaxin. This is called chemotactic deactivation. We developed a new deactivation chamber to determine chemotaxis and chemotactic deactivation of human PMN. This novel chamber is a modification of the commercially available acrylic 48-well microchemotaxis chamber consisting of an upper block with wells drilled all the way through the block and a blind-well lower block. Both blocks are separated by a polycarbonate membrane. PMN from the wells in the upper block migrate through the pores of the membrane into the wells of the lower block containing the chemoattractants. Migrated PMN on the lower side of the PC membrane were quantified after staining by measuring specific light absorbance. The chemotactic activity is quantified as a ratio of stimulated migration and random migration (chemotactic index=CI). For our novel chamber, only the upper blocks of this commercial chamber were connected like a sandwich, including a polyvinylpyrrolidone-free polycarbonate membrane with a pore size of 3 microm. The wells in the upper compartment were filled with 5 x 10(4) PMN and deactivating chemotaxin. The lower block was then filled with the chemotactic stimulus and the chamber was then incubated in humidified air with 5% CO2 atmosphere at 37 degrees C. The influence of cell concentration, incubation time, chemotactic factor concentration, pore size and alkaline treatment of polycarbonate membranes on migrational activity of PMN have been investigated. The technique was rigorously standardized in order to optimize the assay conditions. The method is relatively simple, sensitive and fast. The determination of chemotaxis and deactivation are performed in the same chamber, thus avoiding cell loss due to nonspecific adherence in other incubation tubes. The chamber can be used to characterize the chemotactic activity of chemoattractants of unknown structure via known and unknown receptors. This new chamber can be very helpful in detecting unknown chemotactic stimuli, which are not detectable by, for example, antibodies. PMID- 11384763 TI - Nonlinear electrophoresis and focusing of macromolecules. AB - Effects of nonlinear dependence drift velocity of (double-stranded) DNA vs. electric field strength were investigated. In comparatively weak fields, the molecular drift velocity is proportional to the external electric field, while in strong fields there is additional nonlinear component. This effect offers possibilities to manipulate the total drift velocity at will-the macromolecules of different size can be made to move in opposite directions in pulsed field gel electrophoresis.A new approach for focusing DNA molecules based on nonlinear electrophoresis and geometric trapping in electric fields is proposed. The focusing is carried out in an alternating nonuniform electric field, created by using a wedge gel with hyperbolic boundaries. It is shown that the fractions separated in such wedge retain their rectilinear shape. Gel electrophoresis experiments supported the possibility of a pronounced nonlinear focusing of DNA molecules. This nonlinear separation technique presents encouraging prospects for micromanipulating systems and also for preparative isolation of long DNA fragments and development of new separation methods for bacterial fingerprinting. PMID- 11384764 TI - Quantitative comparison of two types of planar lipid bilayers--folded and painted -with respect to fusion with vesicles. AB - Two major types of planar lipid bilayers, painted and folded, were compared with respect to vesicle fusion using one chamber for the preparation of both bilayers. Liposomes containing ion channels composed of nystatin and ergosterol were used as the vesicle sample. Fusion of the liposome to either bilayer elicited a spike like current change, which corresponds to a fusion event. The lag time between the first fusion event and the addition of the vesicles is an index of the ease with which the vesicles fuse with the bilayers. The lag time in the painted bilayer at a KCl concentration (cis) of 450 mM was 1.58+/-1.18 min, similar to that in the folded bilayer (1.65+/-0.64 min). The lag time decreased with increase of the osmotic difference across the painted bilayer, whereas this change was small in the folded bilayer. The fusion of the liposomes to the painted bilayer was markedly reduced by stopping the stirring in the cis compartment, whereas the fusion to the folded bilayer was not affected significantly. These results imply that no practical difference exists in the ability of vesicles to fuse with the painted and folded bilayers. For the study of single channel behavior, the painted bilayer could have an advantage because simply stopping the stirring prevents excess fusion. PMID- 11384765 TI - Structural analysis of heavy ion radiation-induced chromosome aberrations by atomic force microscopy. AB - Heavy ion radiation (high linear energy transfer, LET, radiation) induces various types of chromosome aberration. In this report, we describe a new method employing an atomic force microscope (AFM) for nanometer-level structural analysis of chromosome damage induced by heavy ion irradiation. Metaphase mouse chromosomes with chromatid gap or chromatid breaks induced by heavy ion irradiation were marked under a light microscope. Then the detailed structure of chromosomes of Giemsa-stained or unstained samples was visualized by the AFM. The height data of chromosomes obtained by AFM provided useful information to distinguish chromatid gaps and breaks. A fibrous structure was observed on the unstained chromosome, the average width of which was about 45.8 nm in the image of AFM. These structures were considered to be 30-nm fibers on the chromosome. The structure of the break point regions induced by neon- or carbon-ion irradiation was imaged by AFM. In some cases, the fibrous structure of break points was detected by AFM imaging after carbon ion irradiation. These observations indicated that AFM is a useful tool for analysis of chromosome aberrations induced by heavy ion radiation. PMID- 11384766 TI - Detection of peptidase activity with 6-aminoquinolyl carbamate-angiotensin I. PMID- 11384767 TI - Central leptin gene therapy suppresses body weight gain, adiposity and serum insulin without affecting food consumption in normal rats: a long-term study. AB - The weight-reducing effects of leptin are predominantly mediated through the hypothalamus in the brain. Gene therapy strategies designed for weight control have so far tested the short-term effect of peripherally delivered viral vectors encoding the leptin gene. In order to circumvent the multiple peripheral effects of hyperleptinemia and to overcome the age-related development of leptin resistance due to multiple factors, including defective leptin transport across the blood brain barrier, we determined whether delivery of viral vectors directly into the brain is a viable therapeutic strategy for long-term weight control in normal wild-type rats. A recombinant adeno-associated virus (rAAV) vector encoding rat leptin (Ob) cDNA was generated (rAAV-betaOb). When administered once intracerebroventricularly (i.c.v.), rAAV-betaOb suppressed the normal time related weight gain for extended periods of time in adult Sprague-Dawley rats. The vector expression was confirmed by immunocytochemical localization of GFP and RT-PCR analysis of leptin in the hypothalamus. This sustained restraint on weight gain was not due to shifts in caloric consumption because food-intake was similar in rAAV-betaOb-treated and rAAV-GFP-treated control rats throughout the experiment. Weight gain suppression, first apparent after 2 weeks, was a result of reduced white fat depots and was accompanied by drastically reduced serum leptin and insulin concentrations in conjunction with normoglycemia. Additionally, there was a marked increase in uncoupling protein-1 (UCP1) mRNA expression in brown adipose tissue, thereby indicating increased energy expenditure through thermogenesis. Seemingly, a selective enhancement in energy expenditure following central delivery of the leptin gene is a viable therapeutic strategy to control the age-related weight gain and provide protection from the accompanying multiple peripheral effects of hyperleptinemia and hyperinsulinemia. PMID- 11384768 TI - Type-2 somatostatin receptor mRNA levels in breast and colon cancer determined by a quantitative RT-PCR assay based on dual label fluorogenic probe and the TaqMan technology. AB - We reported previously that the expression of type 2 somatostatin receptor (sst2) was positively related to patient outcome in the childhood tumor neuroblastoma. To quantitate the expression of mRNA sst2 expression, we used a competitive RT PCR assay. To improve the practicability of this measurement and its applicability to large groups of patients, we present here an original 'real time' quantitative RT-PCR method, based on a dual-labeled fluorogenic probe and the TaqMan technology. By this method, we have measured sst2 mRNA expression in 24 breast cancer samples and 26 colon carcinomas as well as on the corresponding non-adjacent non-neoplastic tissue of the same patients. The proposed method has a dynamic range of 4 x 10(4) to 4 x 10(8) molecules of sst2 mRNA. The intra-assay precision of the test, evaluated as signal detection variability, was 2.4%. Accuracy, evaluated by the addition of standard RNA to unknown samples, provided a mean recovery of 98+/-2%. A significant correlation has been observed in a study performed in 24 neuroblastoma samples measured both with the proposed method and with a competitive RT-PCR assay (r=0.913, p<0.001). In our preliminary clinical study, no significant differences were observed in sst2 mRNA levels between normal and tumor specimens in both colorectal (normal tissue 5.1 x 10(7)+/-2.0 x 10(7) molecules/microg total RNA, cancer tissue 9.7 x 10(7)+/-4.2 x 10(7)) and breast tumors (normal tissue 5.5 x 10(8)+/-2.0 x 10(8), cancer tissue 4.4 x 10(8)+/-3,7 x 10(8)).However, in colorectal cancer, sst2 mRNA values of subjects with high circulating carcinoembryonic antigen (CEA) levels (>5 ng/ml) were statistically lower (2.3 x 10(7)+/-6.2 x 10(6) molecules/, microg total RNA; p<0.05) than those of subjects with low CEA concentration (1.4 x 10(8)+/-6.7 x 10(7)). Also, the sst2 mRNA ratio between normal and tumor tissue (N/T ratio) resulted significantly inversely related to CEA levels. In breast cancer, a significant difference was found between the mean N/T ratio of negative (below 10 fmol/mg protein) and positive estrogen receptor tumors (p<0.05). Analogous results were found selecting breast tumors on the basis of the progesterone receptor status (p<0.05). The proposed method is accurate, precise, sensitive and less labor-intensive than the competitive RT-PCR assay. For a correct evaluation of sst2 mRNA expression, it seems very important to measure the sst2 expression both in tumor and in the non-tumoral non-adjacent tumor specimens. PMID- 11384769 TI - The novel peptide apelin lowers blood pressure via a nitric oxide-dependent mechanism. AB - Apelin is an endogenous ligand of the human orphan receptor APJ. We detected apelin-like immunoreactivity in the adipocytes, gastric mucosa, and Kupffer cells in the liver. We also detected apelin-like immunoreactivity localized within the endothelia of small arteries in various organs. Further, it was found that mean arterial pressure after the administration of apelin-12, apelin-13, and apelin-36 at a dose of 10 nmol/kg in anaesthetized rats was reduced by 26+/-5, 11+/-4, and 5+/-4 mm Hg, respectively. In the presence of a nitric oxide (NO) synthase inhibitor, the effect of apelin-12 on blood pressure was abolished. Furthermore, the administration of apelin-12 (10 nmol/kg) in rats produced a transitory elevation of the plasma nitrite/nitrate concentration from a basal level of 21.4+/-1.6 to 27.0+/-1.5 microM. Thus, apelin may lower blood pressure via a nitric oxide-dependent mechanism. PMID- 11384770 TI - A new angiotensinergic system in the CNS of the rat. AB - The use of two different polyclonal, affinity-purified, monospecific antibodies to ANG II (called BODE and BODE 1) revealed dissimilar distribution of ANG II immunoreactivity within the rat central nervous system (CNS). The ANG II-like material detected using BODE was concentrated in the neurosecretory hypothalamic nuclei, in the inner layer of the median eminence and in the posterior lobe of the pituitary. In contrast, the BODE 1 antibody did not stain the hypothalamic neurohypophysial angiotensinergic system, and the staining pattern was much more broadly distributed throughout the CNS. BODE 1 is the first antibody that can be used to verify the locations of endogenous angiotensin and their receptor sites in the CNS. The diverse distribution of the ANG II-like material detected by the two antibodies provides strong evidence for the existence of at least two different angiotensinergic systems in the CNS. PMID- 11384771 TI - Endogenous cholecystokinin plays a role in down-regulation of pancreatic amylase independent of dietary carbohydrate in rats. AB - The role of cholecystokinin (CCK) in the regulation of pancreatic amylase has not been fully clarified. We examined the effects of hyperCCKemia with chronic pancreatico-biliary diversion (PBD) and blockade of CCK(A)-receptor on rat pancreatic amylase activity and mRNA abundance. Also, we examined the relationship between diet and CCK in terms of regulation of pancreatic amylase. PBD was produced by transposition of the duodenal segment containing the ampulla of Vater to the upper ileum. A potent CCK(A)-receptor antagonist, devazepide, was injected (6 mg/kg body weight per day for 5 days) in the PBD rats fed with diets containing normal or low level of carbohydrate (695 or 345 g sucrose/kg diet). The specific activity and mRNA abundance of the pancreatic amylase were constantly lower 4, 10 and 28 days after PBD than those after the sham operation. Devazepide treatment completely restored the amylase activity lowered by PBD without any increases in amylase mRNA. Feeding a high-protein low-carbohydrate diet suppressed the pancreatic amylase activity and mRNA abundance in PBD rats to a similar degree in those treated, and those untreated, with devazepide. We conclude that endogenous CCK suppresses pancreatic amylase production, and we speculate that CCK reduced translational efficiency of amylase mRNA. The effect of CCK on amylase production is independent of regulation by dietary carbohydrate. PMID- 11384772 TI - Amphibian glucagon family peptides: potent metabolic regulators in fish hepatocytes. AB - Peptides analogous to glucagon-like peptide-1 (GLP-1) have been isolated from amphibian pancreas and intestine, and their amino acid sequences and cDNA structures elucidated. Just like their mammalian counterpart, these peptides are potent insulinotropins in mammalian pancreatic cells. We show here that these peptides also exert strong glycogenolytic actions when applied to dispersed fish hepatocytes. We compared the potencies of three synthetic GLP-1s from Xenopus laevis and two native GLP-1s from Bufo marinus in the activation of glycogenolysis in the hepatocytes of a marine rockfish (Sebastes caurinus) and two freshwater catfish (Ameiurus nebulosus and A. melas), and demonstrated their effectiveness in increasing the degree of phosphorylation of glycogen phosphorylase. We also compared the glycogenolytic potency of the peptides with those of human GLP-1 and glucagons from human and B. marinus. Sensitivity to these peptides is species-specific, with the rockfish responding at lower concentrations to GLP-1s and the two catfish reacting better to glucagons. However, the relative potency of the amphibian GLP-1s and glucagons is similar in the three species. Xenopus GLP-1C (xGLP-1C) is consistently more potent than xGLP 1B, while xGLP-1A displays the smallest activation of glycogenolysis. Similarly, Bufo GLP-1(32)-the peptide with the highest amino acid sequence identity to xGLP 1C-always shows a higher potency than Bufo GLP-1(37), which is closely related to xGLP-1B. The relative hierarchy of these glycogenolytic GLP-1s differs from their ranking as insulinotropins in mammalian beta-cells. In the rockfish system, Bufo glucagon-36, a C-terminally extended glucagon, is more potent than the shorter bovine glucagon and Bufo glucagon-29 in the activation of glycogenolysis; when tested in A. nebulosus hepatocytes, bovine and amphibian glucagons are equipotent. Amphibian GLP-1s and glucagons activate glycogenolysis in fish hepatocytes through increased phosphorylation of glycogen phosphorylase, implying involvement of the adenylyl cyclase/protein kinase A system in signal transduction. We conclude that the broad physiological effectiveness of GLP-1 has been retained throughout vertebrate evolution, and that both insulinotropic activity and glycogenolytic actions belong to the repertoire of GLP-1. PMID- 11384773 TI - Selective down-regulation of AT2 receptors in uterine arteries from pregnant ewes given 24-h intravenous infusions of angiotensin II. AB - Previously, we showed that uterine arteries from late gestation pregnant ewes infused intravenously with angiotensin II (Ang II) for 24 h, displayed heightened responsiveness to Ang II in vitro. Furthermore, we found that a small population of ewes with a "preeclampsia-like" disorder also displayed this. Therefore, we have investigated the density and affinity of Ang II receptor subtypes in the uterine arteries from these groups. Ang II receptor binding was measured using 125I [Sar1Ile8] Ang II. Proportions of AT1 and AT2 receptors were determined by inhibiting 125I [Sar1Ile8] Ang II with losartan (AT1 antagonist) or PD 123319 (AT2 antagonist). Uterine arteries from 24-h Ang II-infused ewes had a lower proportion of AT2 receptors (56.2+/-2.3%) than control (saline-infused) ewes (84.1+/-1.0%; P<0.05). The density of AT2 receptors was reduced (P<0.05) while the density of AT1 receptors was not different. Thus, 24-h infusions of Ang II selectively down-regulated AT2 receptors in the uterine artery, resulting in heightened Ang II reactivity. By contrast, the binding properties of Ang II receptor subtypes in uterine arteries from ewes with the "preeclampsia-like" disorder were not different from control ewes. PMID- 11384774 TI - Inflammatory response to cold injury in remote organs is reduced by corticotropin releasing factor. AB - Current experimental evidence concerning the potential activity of corticotropin releasing factor (CRF) in inflammatory processes still remains controversial. To determine whether CRF has protective effects on three remote organs (liver, lung and stomach) affected by cold injury and to characterize the role of neutrophils in cold-induced inflammation, dorsums of anesthetized rats were exposed for 5 min to a 22% NaCl solution maintained at -20+/-0.5 degrees C and the rats were sacrificed at 24 h after the cold injury. The results indicate that cold-exposure induced edema in the liver, lung and stomach was blocked by subcutaneous (s.c.; 1.2 and 12 nmol/kg; 30 min before cold trauma) CRF pretreatment, while the central administration of CRF (intracisternally (i.c.); 0.30 and 1.5 nmol/rat; 15 min before cold) had the similar effect at the higher dose. Histological assessment and the tissue myeloperoxidase activities also revealed that CRF given peripherally has a protective role in damage generation. Moreover, CRF had a facilitatory effect in the recovery of the body temperature following cold exposure. In conclusion, CRF is likely to act on its peripheral receptors in the inflamed remote organs, suppressing the edematogenic effects of inflammatory mediators, some of which are neutrophil-derived. PMID- 11384775 TI - A-like cells in the rat stomach contain ghrelin and do not operate under gastrin control. AB - Ghrelin is a 28 a.a. gastric peptide, recently identified as a natural ligand of the growth hormone secretagogue receptor (orphan receptor distinct from the receptor for growth hormone releasing hormone). In the present study, radioimmunoassay demonstrated ghrelin-like material in the rat oxyntic mucosa with moderate amounts also in antrum and duodenum. Small amounts were found in the distal intestines and pancreas. Northern blot analysis revealed abundant ghrelin mRNA in the oxyntic mucosa. Immunocytochemistry demonstrated ghrelin immunoreactivity in endocrine-like cells in the oxyntic mucosa. Such cells occurred in low numbers also in the antrum and duodenum. The rat oxyntic mucosa is rich in endocrine (chromogranin A/pancreastatin-immunoreactive) cells, such as the histamine-rich ECL cells (65-75% of the endocrine cells), the A-like cells (20-25%) and the D cells (somatostatin cells) (10%). The ghrelin-immunoreactive (IR) cells contained pancreastatin but differed from ECL cells and D cells by being devoid of histamine-forming enzyme (ECL cell constituent) and somatostatin (D cell constituent). Hence, ghrelin seems to occur in the A-like cells. The ghrelin-IR cells in the antrum were distinct from the gastrin cells, the serotonin-containing enterochromaffin cells and the D cells. Conceivably, ghrelin cells in the antrum and distally in the intestines also belong to the A-like cell population. The concentration of ghrelin in the circulation was lowered by about 80% following the surgical removal of the acid-producing part of the stomach in line with the view that the oxyntic mucosa is the major source of ghrelin. The serum ghrelin concentration was higher in fasted rats than in fed rats; it was reduced upon re-feeding and seemed unaffected by 1-week treatment with the proton pump inhibitor omeprazole, resulting in elevated serum gastrin concentration. Infusion of gastrin-17 for 2 days failed to raise the serum ghrelin concentration. Omeprazole treatment for 10 weeks raised the level of HDC mRNA but not that of ghrelin mRNA or somatostatin mRNA in the oxyntic mucosa. Hence, unlike the ECL cells, ghrelin-containing A-like cells do not seem to operate under gastrin control. PMID- 11384776 TI - Differential alterations in tachykinin NK2 receptors in isolated colonic circular smooth muscle in inflammatory bowel disease and idiopathic chronic constipation. AB - Inflammatory bowel disease (IBD) and idiopathic chronic constipation (ICC) are intestinal disorders which disrupt normal colonic motility. Enteric tachykinins are well-recognised to play a role in the motor control of the gut, and increased colonic levels of substance P are seen in IBD, whereas decreased levels have been reported in ICC. In this investigation, we have characterised the tachykinin receptor population of normal human colonic circular smooth muscle and examined any changes that occur in IBD and ICC. The selective tachykinin NK2 receptor agonist, [beta-Ala8]neurokinin A(4-10), caused concentration-dependent contractions in healthy tissues; neither NK1 receptor-selective nor NK3 receptor selective agonists were contractile. In diseased preparations also, only [beta Ala8]neurokinin A(4-10) caused contractions with EC50 values similar to health. The maximum contractile responses (Emax), however, were significantly decreased in both forms of IBD but significantly increased in ICC. The muscarinic acetylcholine receptor agonist, carbachol, also caused contractions in diseased tissues, but EC50 and Emax values were not significantly different from health. The differential changes in contractility found in IBD and ICC are specific to NK2 receptors, and may reflect the altered levels of substance P or other tachykinins found in these intestinal disorders. PMID- 11384777 TI - Antibodies raised against the extracellular tail of the CCKB/gastrin receptor inhibit gastrin-stimulated signalling. AB - INTRODUCTION: Gastrin acts to stimulate gastric acid secretion and is an acknowledged growth factor for human gastrointestinal (GI) cancer. The identity of the exact receptor type mediating the growth promoting effects of gastrin in tumours is uncertain. However, the best-characterised gastrin receptor is the CCK receptor type B (CCKB)/gastrin receptor. The anti-GRE1 antibody is a polyclonal, affinity-purified antibody raised against GRE1, a synthetic 21 amino acid peptide homologous to part of the extracellular, N-terminal tail of the CCKB receptor. We have recently proven that GRE1 antiserum specifically localises CCKB receptors on CCKB receptor transfected NIH3T3 cells and human gastrointestinal tumour cells by Western blotting and immunocytochemistry. GRE1 antiserum also inhibits liver invasion in the C170HM2 colorectal liver-metastasis model. AIM: To relate the ability of GRE1 antiserum to displace G17 from CCKB receptors with its impact on cellular transduction effects. METHODS: Radioligand binding studies were performed with 125IG17 and Calcium mobilisation studies by use of the fluorescent dye Fura 2-am. RESULTS: GRE1 antiserum competitively displaced 50% radiolabelled gastrin-17 from whole cell NIH3T3 CCKB transfectants at a protein concentration of 250 microg x ml(-1). GRE1 antiserum did not stimulate calcium ion influx in the transfectant NIH3T3 cells when used at a range of protein concentrations. Pre incubation with GRE1 antiserum was required to inhibit gastrin-stimulated calcium ion influx. This was found to be concentration-dependent, with inhibition shown at 30 and 5 microg x ml(-1) but not at 500 ng x ml(-1) or below. CONCLUSION: The GRE1 antiserum is specific for the CCKB receptor and may act to inhibit gastrin stimulated signalling in tumour cells. PMID- 11384778 TI - Effect of calcitonin gene related peptide on the adenylate cyclase activity in abalone gill membranes. AB - The aim of the present study was to investigate and to compare the effect of calciotropic hormones, human calcitonin gene related peptide (CGRP) I and II, salmon calcitonin (CT) and human amylin on the adenylate cyclase activity in abalone gill membranes. In addition to human CGRPI, human CGRPII and salmon CT stimulated the adenylate cyclase activity. No effect was observed with amylin. The higher effect was observed with human CGRPI and II that induced a 160-170% increase of the enzyme activity. Fifty percent of the maximal activity was observed with 3 and 8 nM of CGRP I and II, respectively. Salmon CT induced a lower effect: the maximal activity was obtained with a hormone concentration of 266 nM and represented a 130% stimulation of the basal activity. In the presence of CGRP 8-37, a competitive antagonist of CGRP action, the stimulation observed with CGRPI was abolished and returned to the basal level. This study points out that, in invertebrates, CGRP receptors present in gill membranes are linked to an adenylate cyclase system similar to that described in vertebrates. In addition, these data are in favour of a role for CGRP in branchial function both in non mammalian vertebrates and invertebrates PMID- 11384779 TI - Overexpression of parathyroid hormone-related protein promotes cell growth in the rat intestinal cell line IEC-6. AB - The rat intestinal cell line, IEC-6, was used as a model to study effects of parathyroid hormone-related protein (PTHrP) on crypt cell growth. Studies showed that addition of PTHrP analogs (1-34), (67-86), or (107-139) to growth medium did not affect proliferation of cells grown in either high (10% Nu-Serum) or low serum (1% Nu-Serum). However, studies on clonal lines of IEC-6 cells stably transfected with PTHrP cDNA and overexpressing PTHrP showed that increased PTHrP production enhanced cell growth and 3H-thymidine incorporation in high, but not low, serum. Additional studies examined the role of the nuclear localization sequence (NLS) of PTHrP in mediating the growth effect. In three clonal IEC-6 lines transfected with PTHrP cDNA bearing a mutated NLS, the ability of PTHrP to stimulate 3H-thymidine incorporation and cell growth was lost. The results suggest that endogenously produced PTHrP can promote proliferation of IEC-6 cells and that the integrity of the NLS of PTHrP is required for its growth effects. PMID- 11384780 TI - Angiotensin and its AT2 receptor: new insights into an old system. AB - The AT2 receptor represents a true receptor, but signals and functions in unexpected ways compared to the respective features of the 'classical' AT1 receptor. Moreover, some of the actions of the AT2 receptor are even directly opposed to those of the AT1 receptor, especially concerning the growth- and differentiation-modulating actions of ANG II. The regulation of the AT2 receptor itself by its agonist, as well as by growth factors during ontogenesis, and its acknowledged effects on the regulation of cell growth, differentiation and apoptosis, points towards a role of a program modulator in embryonic development and regeneration. PMID- 11384781 TI - Calcitonin gene-related peptide-mediated ischemic preconditioning in the rat heart: influence of age. AB - In the present study, we examined whether age-related reduction of ischemic preconditioning is related to calcitonin gene-related peptide (CGRP) release in the rat heart. Thirty minutes of global ischemia and 40 min of reperfusion caused a significant decrease of cardiac function and a marked increase of creatine kinase (CK) release at 2, 6 and 20 months of age. Ischemic preconditioning and pretreatment with CGRP for 5 min significantly improved cardiac function and reduced CK release during reperfusion at 2 and 6 months of age but not at 20 months of age. The content of CGRP in the coronary effluent during ischemic preconditioning was significantly increased in the first cycle at 2, 6 months of age but not at 20 months of age. These results suggest that the protection afforded by ischemic preconditioning is decreased in aging hearts, and the age related change may be related to reduction of the release and effect of CGRP in the rat heart. PMID- 11384782 TI - Comparative analysis of the central CCK system in Fawn Hooded and Wistar Kyoto rats: extended localisation of CCK-A receptors throughout the rat brain using a novel radioligand. AB - The neuropeptide cholecystokinin has been implicated in the actions of a number of central processes including anxiety and reward. For this reason, the aim of the present study was to compare the density of CCK-A and -B receptors and the mRNA encoding preproCCK throughout the brains of an alcohol-preferring (Fawn Hooded) rat strain with that of a non-alcohol-preferring (Wistar Kyoto) strain of rat. Our study revealed significant differences with regard to the central CCK system of the FH compared to the WKY rat, including differences in CCK-A receptor binding throughout the dorsal medulla, and altered CCK-B binding density throughout the cerebral cortex and reticular nucleus of the thalamus. The most striking result, given the altered behavioural phenotype of the FH rat, was the 33% lower density of CCKmRNA measured throughout the ventral tegmental area of the FH rat when compared to the WKY. This study also reports on a protocol to utilise a novel radioligand, [125I]-D-Tyr-Gly-A-71378, for autoradiographic detection of CCK-A receptors throughout the rat brain. As previously reported, CCK-A receptors were located throughout the area postrema, interpeduncular nucleus and nucleus tractus solitarii; however, binding to CCK-A receptors was also visualised throughout the medial pre-optic area, the arcuate nucleus and the circumventricular regions of the ventral hypothalamus, regions known to contain CCK-A receptors but which were previously undetectable using autoradiography in rat brain. PMID- 11384783 TI - Circulating immunoreactive proANP1-30 and proANP31-67 responses to acute exercise. AB - The circulating immunoreactive atrial natriuretic peptide (C-terminal; alpha-ANP) increases during exercise to become suppressed in the first hours of the recovery. The response of the N-terminal ANP fragments to acute exercise is not known while proANP (31-67) appears to be elevated with chronic exercise. We evaluated the plasma concentrations of the N-terminal ANP fragments (1-30) and (31-67) in oarsmen (n=10) before and after two acute exercise bouts separated by 5 h. As control, measurements were made on a day with no exercise (n=12). At rest, the concentrations of proANP(1-30) and proANP(31-67) were 344+/-42 and 810+/-172 pmol x l(-1), respectively. Half an hour after the first exercise bout, proANP(1-30) was elevated (to 404+/-48 pmol x l(-1); P<0.05) and decreased below the pre-exercise level (to 316+/-41 pmol x l(-1); P<0.05) 4 h into the recovery period. Also, 30 min after the second exercise session, the concentration of proANP(1-30) was elevated to 408+/-45 pmol x l(-1) (P<0.05) and the pre-exercise level was re-established on the following morning. Thus, proANP(1-30), rather than proANP(31-67), responded to acute exercise. These results suggest that atrial distension and, therefore, the central blood volume changes markedly in athletes during a day with repeated exercise bouts. PMID- 11384784 TI - Genetic deletion of angiotensin AT2 receptor leads to increased cell numbers in different brain structures of mice. AB - Angiotensin II (Ang II) is a potent vasoactive peptide and displays growth factor like properties. Different high-affinity Ang II receptor subtypes (AT1A, AT1B and AT2) have been cloned. They are expressed in various brain structures. Additionally, it has been assumed that Mas could interact directly or indirectly with the renin-angiotensin system. The AT1 receptor mediates pressor and mitogenic effects of Ang II, whereas physiological function and signaling mechanisms of the AT2 receptor remain poorly understood. Recent reports have shown that Ang II could mediate apoptosis through AT2 receptors. Since the AT1A, AT2 and Mas knockout mice provide new tools for uncovering potential actions of Ang II, the cell number in different brain structures of male adult wild-type mice and mice deficient for AT1A, AT2 or Mas was evaluated to get more insight into the role of Ang II in central nervous system development. In nearly all investigated brain structures (cortex, hippocampus, amygdala, thalamus), the cell number was significantly higher in AT2-deficient mice in comparison to wild-type mice. To the contrary, in AT1A-deficient mice the cell number was significantly less than in controls in the lateral geniculate and the medial amygdaloid nucleus. However, cell numbers were not changed in Mas-knockout mice compared to their wild-types. These results show the contrary effects of both angiotensin receptors on cell growth and represent the first demonstration of their action on neuronal cell development evidenced in the adult mouse brain. PMID- 11384785 TI - Adaptive designs, informed consent, and the ethics of research. AB - The ethical tension in research design is often characterized as that between individual and collective ethics. While adaptive clinical trials (ACTs) are generally considered to be more sensitive to individual ethics, the concomitant loss of statistical power associated with them is often used to justify randomized clinical trials (RCTs). This paper challenges this characterization of the central ethical problem in research design. It argues that the key consideration in clinical research hinges on the process of informed consent. When the research context is such that the subject is able to provide informed consent, RCTs can be justified and may be required. However, in desperate medical situations the process of informed consent is often undermined. It is argued that in such situations ACTs are ethically required. We introduce "the principle of interchangeability" and argue that it must be satisfied if research in desperate medical situations is to be justified. PMID- 11384786 TI - Sample size and power calculations with correlated binary data. AB - Correlated binary data are common in biomedical studies. Such data can be analyzed using Liang and Zeger's generalized estimating equations (GEE) approach. An attractive point of the GEE approach is that one can use a misspecified working correlation matrix, such as the working independence model (i.e., the identity matrix), and draw (asymptotically) valid statistical inference by using the so-called robust or sandwich variance estimator. In this article we derive some explicit formulas for sample size and power calculations under various common situations. The given formulas are based on using the robust variance estimator in GEE. We believe that these formulas will facilitate the practice in planning two-arm clinical trials with correlated binary outcome data. PMID- 11384787 TI - Analysis, sample size, and power for estimating incremental net health benefit from clinical trial data. AB - Stinnett and Mullahy recently introduced the concept of net health benefit as an alternative to cost-effectiveness ratios for the statistical analysis of patient level data on the costs and health effects of competing interventions. Net health benefit addresses a number of problems associated with cost-effectiveness ratios by assuming a value for the willingness-to-pay for a unit of effectiveness. We extend the concept of net health benefit to demonstrate that standard statistical procedures can be used for the analysis, power, and sample size determinations of cost-effectiveness data. We also show that by varying the value of the willingness-to-pay, the point estimate and confidence interval for the incremental cost-effectiveness ratio can be determined. An example is provided. PMID- 11384788 TI - Locating study subjects: predictors and successful search strategies with inmates released from a U.S. county jail. AB - Minimizing loss to follow-up in longitudinal studies is critical. The purpose of this study was to examine the ability to locate subjects recently released from jail, identify predictors of being able to find a subject, and describe effective search strategies for this unique population. The sample for this cohort study included study subjects who were sought for interview after release from jail. Inmates in the San Francisco City and County Jail were enrolled in a randomized trial of incentives to improve follow-up for tuberculosis therapy after release from jail. Sociodemographic, health-related, and extensive locating information was collected during baseline interviews in jail. The main outcome was successful location of the subject. Study personnel recorded data on the number and nature of attempts made to find subjects in order to describe successful search strategies. Of 254 persons sought for the postrelease interview, 188 (74.0%) were found. Primary English speakers were more likely than Spanish speakers to be found (relative risk: 3.2, 95% confidence interval: 1.5-6.7, p = 0.002). Nearly one quarter of subjects (24%) were found back in jail, and the remainder were found in the community. Phone calls and letters to the subjects, and personal contacts to family and friends were successful strategies for 53% of the subjects. Seeking persons in programs, such as shelters and drug and alcohol programs, was successful in finding 18% of English-speaking subjects. Outreach efforts in sections of the city where Latinos spent time, including popular restaurants and community gathering places, were successful in finding 13% of Spanish-speaking subjects. We conclude that study subjects released from jails can be successfully located using well-defined search protocols tailored to the ethnicity of the sample and including a variety of strategies. Employment of bilingual personnel is important when a large proportion of subjects is monolingual and non-English speaking. PMID- 11384789 TI - Cause-specific mortality coding. methods in the collaborative ocular melanoma study coms report no. 14. AB - Ascertainment of cause of death is often sought in clinical trials in which mortality is an outcome of interest. Standardized methods of coding all-cause and disease-specific mortality were developed and evaluated in the Collaborative Ocular Melanoma Study randomized trial of pre-enucleation radiation of large choroidal melanoma. All available clinical and pathologic materials documenting events prior to each reported death were reviewed systematically by a Mortality Coding Committee (MCC) to determine whether melanoma metastasis or local recurrence was present at the time of death. A level of certainty was assigned based on availability of local or central review of pathology materials. The outcome of the mortality coding protocol was evaluated both by assessing agreement between the judgment of the MCC and the presumed cause of death reported by the clinical center and, for a subset of patients, by assessing agreement between the MCC classification and the cause of death reported on the death certificate. As of July 31, 1997 (the cutoff date for the initial mortality report), 435 (95%) of 457 deceased patient files had been reviewed. The MCC classified 269 patients (62%) as dead with melanoma metastasis, 22 (5%) as dead with another malignant tumor, and 92 (21%) as dead with a malignant tumor of uncertain origin. Thirty-eight patients (9%) died with no evidence of malignancy; in 14 cases (3%), the presence or absence of malignancy could not be established due to lack of clinical information. Fair agreement (kappa = 0.34) was observed between the determinations of the MCC based on detailed review of materials and the cause of death reported on the death certificate, but death certificates alone underestimated the proportion of deaths due to metastatic choroidal melanoma. Detailed mortality coding identified difficulties associated with accurate reporting of cause-specific mortality in patients with choroidal melanoma. PMID- 11384790 TI - Promoting adherence and retention to clinical trials in special populations: a women's health initiative workshop. AB - This paper describes a Women's Health Initiative workshop on promoting adherence and retention in randomized clinical trials among ethnic minority women, participants of lower socioeconomic status, and older women. Workshop objectives were: (1) to increase knowledge of demographic and cultural characteristics of diverse groups, (2) to increase awareness of how diversity can affect interactions in clinical research, (3) to explore how research staff behavior can influence adherence and retention, and (4) to increase knowledge of strategies to enhance adherence and retention in special populations. The workshop emphasized the importance of understanding beliefs, values, and experiences that are common in diverse groups of individuals, while at the same time recognizing and respecting individual differences that result from varying life circumstances and experiences. We discuss strategies to increase cultural competence, reduce stereotypes and discrimination, and create a culturally relevant and sensitive research environment. PMID- 11384791 TI - The prehospital treatment of status epilepticus (PHTSE) study: design and methodology. AB - Status epilepticus is a neurological emergency that is typically first encountered and managed in the prehospital environment. Although aggressive pharmacological treatment of status epilepticus is well established in the emergency department and hospital settings, the relative risks and benefits of active therapy for status epilepticus in the prehospital setting are not known. The Prehospital Treatment of Status Epilepticus (PHTSE) study is a prospective, randomized, double-blind, placebo-controlled study designed to address the following aims: (1) to determine whether administration of benzodiazepines by paramedics is an effective and safe means of treating status epilepticus in the prehospital setting and whether this therapy influences longer-term patient outcome, (2) to determine whether lorazepam is superior to diazepam for the treatment of status epilepticus in the prehospital setting, and (3) to determine whether control of status epilepticus prior to arrival to the emergency department influences patient disposition. The initial phase of the PHTSE study began in January 1994 and was completed in February 1999 after the successful enrollment of 205 patients into the three treatment arms. In this paper, we describe the rationale for the conceptualization of the study and details of the study design and methodology, and emphasize some aspects of study implementation that are unique to research involving the emergency medical system. PMID- 11384792 TI - A multicenter two by two factorial trial of cognitive behavioral therapy and aerobic exercise for Gulf War veterans' illnesses: design of a veterans affairs cooperative study (CSP #470). AB - The Department of Veterans Affairs (VA) Cooperative Studies Program (CSP) Study #470 is a 2 x 2 factorial trial designed to evaluate the hypothesis that both cognitive behavioral therapy (CBT) and aerobic exercise will significantly improve physical function in participants with Gulf War veterans' illnesses (GWVI), and that adding CBT to aerobic exercise will provide further incremental benefit. One thousand three hundred fifty-six veterans will be randomized to one of four treatment arms: CBT plus aerobic exercise plus usual and customary care, aerobic exercise plus usual and customary care, CBT plus usual and customary care, or usual and customary care alone. The study duration is 2.5 years with 1.5 years of intake and 1 year of follow-up. The primary outcome measure is the proportion of veterans improved more than seven units on the physical component summary (PCS) scale of the Short Form Health Survey for Veterans (SF-36V) measured 12 months after randomization. This generic quality-of-life measure was chosen because there is no disease-specific measure for GWVI and the symptoms of GWVI span a wide range of physical manifestations that are related to the domains covered by the PCS scale. Sample size was determined to detect all six pairwise comparisons between the four treatment arms with 90% power and a Bonferroni adjustment for an overall type I error of 0.05 or 0.05/6 = 0.0083. CSP #470 was initiated in May 1999 in 18 VA and two Department of Defense medical centers. To date this represents the largest randomized trial designed to evaluate treatments for individuals with unexplained physical symptoms. This paper will focus on the rationale and unique features of the study design. Control Clin Trials 2001;22:310-332 PMID- 11384793 TI - The childhood asthma prevention study (CAPS): design and research protocol of a randomized trial for the primary prevention of asthma. AB - The Childhood Asthma Prevention Study is a randomized controlled trial to measure whether the incidence of atopy and asthma can be reduced by house dust mite allergen reduction, a diet supplemented with omega-3 fatty acids, or a combination of both interventions. Six hundred and sixteen pregnant women whose unborn children were at high risk of developing asthma because of a family history were randomized prenatally. Study groups are as follows: Group A (placebo diet intervention, no house dust mite reduction), Group B (placebo diet intervention, active house dust mite reduction), Group C (active diet intervention, no house dust mite reduction), and Group D (active diet intervention, active house dust mite reduction). The house dust mite reduction intervention comprises use of physical and chemical methods to reduce allergen contact. The dietary intervention comprises use of a daily oil supplement from 6 months or at onset of bottle-feeding, and use of margarine and cooking oils based on sunflower or canola oils to increase omega-3 dietary intake. Data is collected quarterly until the infant is 1 year old and then half yearly until age 5 years. Questionnaires are used to collect respiratory illness history and information about diet and home environment. Dust is collected from the child's bed and bedroom and playroom floors. Blinded assessments are conducted at 18 months, 3 years, and 5 years. Skin prick tests to common allergens, blood tests, and detailed illness, medication use, and vaccination histories are collected. Primary outcomes will be the development of allergic sensitization and the presence and severity of asthma. This study is designed to measure the effectiveness of allergen reduction and dietary supplementation, both separately and in combination, for the primary prevention of atopy and asthma. The results of this study may have important implications for public health policies to reduce the incidence of childhood asthma. Control Clin Trials 2001;22:333-354 PMID- 11384797 TI - Coeliac disease and reproductive disorders: a neglected association. AB - Coeliac disease is a chronic disease caused by a permanent intolerance to ingested gluten resulting in immunologically mediated inflammatory damage of the small-intestinal mucosa. The wide spectrum of clinical symptoms is partly due to the malnourished state caused by the malabsorption of macro- and micronutrients. Fertility problems, sexual dysfunction and obstetrical complications are more frequently observed in patients with coeliac disease. These reproductive disorders may be a consequence of the endocrine derangements caused by selective nutrient deficiencies. Nowadays, the early diagnosis and treatment of coeliac disease is possible and not very costly. Therefore, coeliac disease must be seriously considered in the preconceptional screening and treatment of patients with reproductive disorders. PMID- 11384798 TI - Umbilical cord morphology and pregnancy outcome. AB - Traditionally, the prenatal assessment of the umbilical cord (UC) is limited to the assessment of the number of vessels and to the evaluation of umbilical artery blood flow parameters. Morphologic aspects of the UC have usually been studies by pathologists and retrospectively correlated with the perinatal outcome. The introduction of more sophisticated imaging techniques have offered the possibility to investigate the UC characteristics during fetal life from early to late gestation. A number of investigations have demonstrated that an altered structure of the UC can be associated with pathologic conditions (i.e. Preeclampsia, fetal growth restriction, diabetes, fetal demise). Nomograms of the various UC components have been generated and allow the identification of lean or large umbilical cords, entities frequently associated with fetal growth abnormalities and diabetes. A Wharton's jelly reduction has also been invoked as a possible cause of fetal death in the presence of single umbilical artery. Prenatal morphometric UC characteristics as well as arterial and venous blood flow parameters in normal and pathologic conditions will be discussed. PMID- 11384799 TI - Vaginal birth after caesarean section in a population with a low overall caesarean section rate. AB - OBJECTIVE: To determine the clinical outcome of vaginal birth after caesarean section (VBAC) in a Dutch population with a low overall caesarean section (CS) rate of 6.5%. STUDY DESIGN: Prospective population based cohort study of 252 patients with a previous caesarean section (CS). Outcome parameters were trial of labour (TOL), success rate and VBAC rate. RESULTS: The TOL rate in the study cohort was 73%, success rate 77%, VBAC rate 56%. The reason for the previous CS influenced success rate. Complications, morbidity and mortality were not different between elective, emergency CS and TOL group, except for a higher incidence of haemorrhage more than 500 ml in the elective CS compared to the TOL group (29% versus 17%, relative risk (RR) 1.74 (1.15--2.34)). CONCLUSIONS: In this Dutch study the success rate is comparable to rate in US study reports. Increase of the VBAC rate can mainly be achieved by increasing the number of women attempting TOL. PMID- 11384800 TI - Term breech and long-term morbidity -- cesarean section versus vaginal breech delivery. AB - STUDY: Perinatal morbidity and mortality of term fetuses have been discussed extensively both for vaginal breech delivery and cesarean section. However, information regarding long-term morbidity and psychomotoric development of these children are scarce. DESIGN: Data of 154 children delivered after breech presentation at our institution between 1988 and 1994 were analyzed using a specific, standardized questionnaire (Enzephalopathiefragebogen, Meyer-Probst) with emphasis on psychomotoric development and skills. Hyperkinetic disorders, social adaptation, emotional instability, and intelligence were evaluated as subcategories and compared to perinatal data. RESULTS: pO(2) and base excess (BE) in the umbilical artery were lower in the vaginal group. pH, body weight and placental weight showed no difference between groups. Psychomotoric development and skills did not differ between children delivered vaginally or abdominally. Perinatal variables did not allow a prediction of long-term morbidity. CONCLUSION: Route of delivery has negligible influence on the measured values in the umbilical artery and no influence on long-term morbidity of fetuses presenting breech. PMID- 11384801 TI - Expectant management of severe pre-eclampsia in the mid-trimester. AB - OBJECTIVE: To determine maternal and perinatal outcomes with expectant management of severe pre-eclampsia in the mid-trimester, using a defined entry point. DESIGN: Prospective case series. Thirty-nine women admitted from 24 to 27 week's gestation with severe pre-eclampsia, whose pregnancies were otherwise stable, were managed expectantly with careful clinical and biochemical monitoring of maternal and foetal status, together with careful blood pressure control, in a high-care obstetric ward. The aim was to safely prolong the pregnancies and thereby improve perinatal outcome. RESULTS: Gestation was prolonged by a median of 12 (range 3--47) days, with greater periods gained at earlier gestations. The overall perinatal loss was 26% and the neonatal loss 17%. The rates of significant maternal complications were low. CONCLUSION: Expectant management of selected women with severe pre-eclampsia from 24 to 27 weeks' gestation in a tertiary care unit is acceptably safe and improves perinatal outcome. PMID- 11384802 TI - The role of magnetic resonance imaging in prenatal diagnosis of fetal anomalies. AB - Magnetic resonance imaging (MR) has become a useful adjuvant in evaluating fetal structural anomalies when ultrasound (US) is equivocal. It has a significant promise in confirming a US suspected abnormality and providing new information that was previously not available. The first studies on prenatal MR were hindered by fetal motion and long acquisition times. This degraded imaging and, therefore, maternal or fetal sedation was needed. Since fast and ultrafast MR with scan times of <1 s have become available, the amount of motion artifacts is decreased and sedation is no longer needed.MR has proved to be especially beneficial in detecting CNS anomalies. Agenesis of the corpus callosum, migration abnormalities and abnormalities of the posterior fossa are better seen on MR. Masses in the fetal neck and thorax can be identified on MR, as some abdominal anomalies. However, the fetal skeletal is difficult to visualize with MR. In the future, it is most likely that real time MR will become clinically available which would improve MR imaging even more. PMID- 11384803 TI - Intrapartum fetal heart rate monitoring: do Swiss midwives implement evidence into practice? AB - OBJECTIVE: Evidence Based Medicine has shown that the results of continuous electronic fetal heart rate (FHR) monitoring are equivalent to those of intermittent auscultation. We were interested in the method midwives preferred to use to monitor FHR during labour and the factors which influenced their choice of method. STUDY DESIGN: A questionnaire with Likert scaled questions was sent to 500 German speaking Swiss midwives. Data analysis was performed by using SPSS for Windows. RESULTS: The majority of the midwives were confident monitoring FHR using intermittent auscultation as their main method during low risk delivery. The essential factors influencing the choice of method of FHR monitoring were their own personal experience and hospital guidelines. Less important were factors such as risk category, litigation, the mother's preferences, research results, time and staffing levels. CONCLUSIONS: Although the skills necessary to implement evidence into obstetrical practice are still available, evidence based research results do not seem to be of great importance, when midwives decide which method to use for intrapartum FHR monitoring. Hospital policies and the professional training received were more important factors. Programs designed to implement evidence in care should reflect the identification and use of these factors in order to facilitate the process of realisation. PMID- 11384804 TI - The role of tamoxifen in the treatment of symptomatic uterine leiomyomata -- a pilot study. AB - OBJECTIVES: To determine the efficacy of tamoxifen in patients with leiomyomata complaining of abdominal pains and vaginal bleeding. STUDY DESIGN: Prospective, randomized, double blind study. A total of 10 patients received for 6 months 20 mg tamoxifen daily, and 10 women received placebo. All patients underwent serial pelvic and ultrasound exams and endometrial sampling was performed prior to initiation of treatment. After 5 years, the patients were interviewed again. RESULTS: Uterine size was not affected by the use of tamoxifen. Patients reported a blood loss decrease of 40--50% at the end of the study (P=0.0001). In the control group a slight increase in blood loss was reported. Hemoglobin levels remained unchanged in both groups. In the study group patients reported after 4 months of treatment a substantial decrease in the intensity of pain (P=0.018). Seven patients in the study group and one patient in the control group developed ovarian cysts. CONCLUSIONS: Treatment with tamoxifen added only marginal benefit while causing unacceptable side effects. Tamoxifen does not seem to be a useful adjunct in the treatment of symptomatic uterine leiomyomata and its use for this indication should be discouraged. PMID- 11384805 TI - Intravenous albumin versus hydroxyethyl starch for the prevention of ovarian hyperstimulation in an in-vitro fertilization programme: a prospective randomized placebo controlled study. AB - A prospective randomized placebo controlled clinical trial was carried out on 250 patients (cycles) considered at risk of developing OHSS in an IVF programme. Criteria for inclusion were: estradiol value of more than 3000 pg/ml or the presence of more than 20 follicles on the day of hCG administration. Patients were randomized by using a random table to receive either 20% human albumin 50 ml (n: 82); 6% hydroxyethyl starch (200/0.5) 500 ml (n: 85) or a placebo of 500 ml 0.9% NaCl solution (n: 83) over 30 min during oocyte collection. Groups were similar with respect to patients' age, estradiol levels on hCG day, body mass index, number of oocytes retrieved, number of embryos transferred and pregnancies (P>0.05). There was no severe OHSS in patients who received albumin and HES while four patients who received placebo developed severe OHSS. On the other hand moderate OHSS was encountered in four patients in the albumin group; five patients receiving HES; and 12 patients receiving placebo. There was a statistically significant difference in the incidence of moderate, severe and overall OHSS among groups (P values of <0.05, <0.05, and <0.01, respectively). Both HES and albumin significantly reduced the incidence of moderate, severe and overall incidence of OHSS. It is concluded that hydroxyethyl starch is a cheaper and safer alternative to Human Albumin in OHSS prevention. PMID- 11384806 TI - In vitro fertilization and blastocyst transfer for carriers of chromosomal translocation. AB - Blastocyst transfers (BT), may benefit chromosomal translocation-carrier couples who suffer multiple miscarriages or are unable to achieve pregnancy following classical ART techniques. In vitro culture applies an additional selection pressure, so that those embryos which achieve blastocyst formation have higher survival probability as healthy balanced translocation carriers or unaffected embryos. Sixteen IVF cycles were performed in 11 patients. When blastocyst are obtained, implantation rate per blastocyst and delivery rates (7/11 cycles, eight healthy babies born) are high. However, the overall blastocyst formation rate is low (20%), and as a consequence in nearly half of the cycles, no blastocyst can be obtained. We propose that this strategy may be used initially as an alternative or a complement to preimplantation genetic diagnosis, and to apply the forces of natural selection in vitro. PMID- 11384807 TI - Plasma levels of free vascular endothelial growth factor(165) (VEGF(165)) are not elevated during gonadotropin stimulation in in vitro fertilization (IVF) patients developing ovarian hyperstimulation syndrome (OHSS): results of a prospective cohort study with matched controls. AB - OBJECTIVE: To evaluate whether differences in plasma vascular endothelial growth factor(165) (VEGF(165)) concentrations exist during gonadotropin stimulation in IVF patients developing severe ovarian hyperstimulation syndrome (OHSS) compared to matched controls. STUDY DESIGN: Prospective cohort study with comparison of 15 OHSS cases with 30 matched (age, follicle numbers, pregnancy) controls. Unpaired students t-test was used to evaluate differences between the OHSS and control group and correlations were calculated with Pearson's test. RESULT(S): Plasma levels of VEGF(165), at four time-points from start of gonadotropin stimulation to embryo transfer (ET), were compared and related to steroid levels and ultrasound data. There were no differences between OHSS and control patients in plasma VEGF(165) levels at any of the four time points, which were compared. The mean levels were between 53--83 pg ml(-1) and 64--83 pg ml(-1) in the OHSS and control group, respectively. Positive correlations existed between total number of follicles, number of large (>15 mm) follicles and VEGF(165) at day of oocyte aspiration and between VEGF(165) and progesterone at ET in the control group, but not in the OHSS group. CONCLUSION(S): Patients developing OHSS do not have raised plasma VEGF(165) levels during gonadotropin stimulation. The lack of positive correlation between VEGF(165) levels and follicle numbers/progesterone in the OHSS group, suggests a disruption in OHSS of the normal controlled follicular VEGF expression. PMID- 11384808 TI - Expression of glutathione S-transferase pi (GST-pi) in human malignant ovarian tumors. AB - OBJECTIVES: In recent years, glutathione S-transferase pi (GST-pi) has attracted much attention and has been studied as a mechanism of multidrug resistance of tumors to anticancer drugs. In the present study, we immunohistologically measured the expression of GST-pi in tumor tissues using surgical specimens obtained from patients with malignant ovarian tumors. METHODS: Of 137 patients with malignant ovarian tumors treated and managed during a period of 20 years since the establishment of Tsukuba University Hospital, 117 patients were selected as subjects because of the presence of complete data on their clinical courses as well as paraffin blocks preserved in a good condition. GST-pi in these specimens was immunohistochemically stained to determine the correlation between GST-pi stainability and clinical outcomes. Stainability was graded as 0 when GST pi was completely absent, 1 when less than 20% of tumor cells were stained, 2 when 20--60% were stained, and 3 when more than 60% were stained. RESULTS: When the correlation between stainability and clinical outcomes was analyzed with Kaplan--Meier method, excluding stage Ia cases that did not receive adjuvant chemotherapy at our hospital, significantly better clinical outcomes were observed in the low stainable group, compared with the high stainable group (P<0.01--0.05, Cox--Mantel test, Wilcoxon's test). CONCLUSION: Since the stainability for GST-pi was high in tumors of histological types with strong resistance to anticancer drugs, and better clinical outcomes were observed in cases having a lower stainability score, the expression of GST-pi was thought to play some role in the resistance of malignant ovarian tumors to anticancer drugs. PMID- 11384809 TI - Ethical guidelines in the prevention of iatrogenic multiple pregnancy. PMID- 11384810 TI - Reasons why women have induced abortions in a developing country. AB - OBJECTIVE: The immediate explanation that women often give for seeking induced abortion is that the pregnancy was unplanned or unwanted. However, the myriad social, economic and health circumstances that underlie such explanations have not yet been fully explored. We wanted to evaluate these factors that lead one to have an abortion. STUDY DESIGN: This prospective study was performed on 588 women applied to our family planning unit to obtain an abortion. Analysis is based on the records of the women's age, educational level, their contraceptive usage and the reasons that they give for having an abortion. We also examined how these reasons were related to women's age, educational attainment and contraceptive choices. RESULTS: The desire to postpone a birth or to stop childbearing is a very common reason given by the women seeking abortion. It is seen that despite the women's desire to postpone or stop childbearing, the majority of them were not using an effective contraceptive method. Women's age is closely related to their reasons for seeking an abortion. On the other hand, we have found no clear association between women's educational level and their main reasons for seeking an abortion. CONCLUSION: Reasons women give for why they seek abortion are often far more complex than simply not intending to become pregnant. While improved contraceptive use can help reduce the unintended pregnancy and abortion, some abortions will remain difficult to prevent, because of limits to women's ability to determine and control all circumstances of their lives. PMID- 11384811 TI - Effect of physicians opinion on patients' choice of treatment. AB - BACKGROUND: While it is obvious that patients should be involved in the decision making process, questions arise as to whether it is feasible to inform them objectively. OBJECTIVE: To assess the impact of the physician's opinion, on the choice of treatment, to the patient. SETTING: We imagined a hypothetical case of a 55-years-old woman with a high risk of osteoporosis. Two patient information letters were written, containing valid scientific information, but reflecting two diverging physicians' opinions, giving the choice between two available medications for osteoporosis prevention: hormone replacement therapy (HRT) and a selective estrogen receptor modulator (SERM), raloxifene. SUBJECTS: We submitted one of the letters (by lottery) to 58 nurses. They were asked to act as this potential patient, and to choose which treatment they preferred to have prescribed. RESULTS: 23/30 women receiving the version favouring the use of HRT, preferred HRT, 26/28 receiving the version favouring SERM preferred this treatment (Chi-Square Pearson 37.3; P<0.001). CONCLUSION: The content of the information provided and its wording have a great influence on the patients' decision. PMID- 11384812 TI - Uterine rupture in second trimester abortion in a grand multiparous woman. A complication of misoprostol and oxytocin. AB - Rupture of unscarred uterus during the second trimester is rare. There have been only 32 cases reported in the literature since 1968. A case of ruptured uterus in a grand multiparous woman is presented. To our knowledge, this might be the first reported case in the English literature of uterine rupture during second trimester termination of pregnancy using a prostaglandin E1 analogue (Misoprostol) and oxytocin. PMID- 11384813 TI - Expectant management of placenta accreta following stillbirth at term: a case report. AB - Placenta accreta is a rare complication of pregnancy with high rates of morbidity and mortality. We report a case of expectant management. This strategy may prevent catastrophic postpartum haemorrhage requiring peripartum hysterectomy. PMID- 11384814 TI - Spontaneous expulsion of a uterine submucosal leiomyoma after administration of a gonadotropin-releasing hormone agonist. AB - A 40-year-old primigravida presented with acute urine retention. Ultrasound examination revealed a large uterine submucosal leiomyoma and GnRH-a was administered. The leiomyoma grew to over twice its original size and protruded through the introitus. After 10 days, it was expelled completely and removed by resectohysteroscopy. The expulsion of the leiomyoma was most likely a result of GnRH-a's flare-up effect. PMID- 11384815 TI - Mitral balloon valvuloplasty during pregnancy in developing countries. AB - Two women presented with severe mitral stenosis at 28th and 23rd weeks of gestation, respectively. Both did not respond to medical therapy and percutaneous balloon valvuloplasty was performed during pregnancy successfully. While one of the women gave no history of cardiac disease the other had already underwent balloon valvuloplasty twice due to restenosis of the mitral valve. The rest of their pregnancies were both uncomplicated. They discontinued their medication. Both were able to deliver vaginally at term. Percutaneous balloon valvuloplasty is a promising approach to the treatment of patients with rheumatic mitral stenosis if medical management is unsuccessful. PMID- 11384816 TI - Management of a triplet pregnancy with two anencephalic fetuses and polyhydramnios. AB - The occurrence of a triplet pregnancy discordant for anencephaly is rare and its management presents a clinical dilemma. We report what appears to be the first case of a triplet pregnancy with two anencephalic fetuses complicated by premature contractions and severe polyhydramnios. Its management, which results a healthy newborn weighing 1385 g is discussed. PMID- 11384817 TI - Wilson's disease in pregnancy: five successful consecutive pregnancies of the same woman. AB - Wilson's disease is an autosomal recessive disorder of copper metabolism characterized mainly by liver cirrhosis and neurological disorders. Appropriate treatment with chelating agents allows normal fertility function. We report five consecutive successful pregnancies of the same woman, treated in the high-risk unit at our medical center. The management dilemmas and treatment options are discussed. PMID- 11384818 TI - First trimester diagnosis of holoprosencephaly and cyclopia with triploidy by transvaginal three-dimensional ultrasonography. AB - We present the prenatal three-dimensional (3D) ultrasound findings in a case of holoprosencephaly and cyclopia at 11 weeks gestation. Only holoprosencephaly with missing cyclopia were initially diagnosed because suboptimal views of the fetal face were obtained with transvaginal two-dimensional (2D) ultrasonography due to fetal position. Chromosomes identified by analysis of a fluid sample from early amniocentesis demonstrated a triploidy (69, XXX), spontaneous fetal demise occurred at 12 weeks and the pregnancy was terminated. This case demonstrated the usefulness of transvaginal 3D ultrasonography in establishing the final diagnosis. PMID- 11384820 TI - Rehabilitation outcomes following percutaneous coronary interventions (PCI). AB - This prospective study evaluated the effect of an individualized, comprehensive, home-based cardiac rehabilitation program combining exercise training with risk factor modification and psychosocial counseling on risk factors, psychological well-being, functional capacity, and work resumption in 99 post-percutaneous coronary interventions (PCI) patients randomized to control (standard care plus telephone follow-up, n=49) or intervention (individualized, comprehensive, home based cardiac rehabilitation, n=50) groups. Data were collected at time 1 (T(1)) during hospital admission, time 2 (T(2)) approximately 2 months post-PCI, and time 3 (T(3)) approximately 12 months post-PCI. Results suggest that the allocation to an individualized, comprehensive, home-based cardiac rehabilitation program provided more advantageous outcomes. At both follow-ups, the intervention group showed within-group improvement in serum cholesterol levels (P<0.02; P<0.01) and exercise participation (P<0.001; P<0.001) with differences in exercise participation favoring the intervention group (P<0.01) at T(2). Repeated measures ANOVA showed significant improvements over time in body mass index (BMI) (P<0.01), psychological well-being (P<0.001), and functional capacity (P<0.001) for both groups. More patients in the intervention group had returned to work at T(2) (P<0.001) and did so more quickly (P<0.01). These findings suggest that an individualized, comprehensive, home-based cardiac rehabilitation program improves risk factor profiles and work resumption patterns for patients following PCI. PMID- 11384821 TI - Using pictographs to enhance recall of spoken medical instructions II. AB - The first study in this series [Houts PS, Bachrach R, Witmer JT, Tringali CA, Bucher JA, Localio RA. Patient Educ. Couns. 1998;35:83-8] found that recall of spoken medical instructions averaged 14% but that, when pictographs (drawings representing the instructions) accompanied the spoken instructions and were present during recall, 85% of medical instructions were remembered correctly. Those findings suggested that spoken instructions plus pictographs may be a way to give people with low literacy skills access to medical information that is normally available only in written form. However, there were three important limitations to that study: (1) the subjects were literate and perhaps literate people remember pictograph meanings better than people with low literacy skills; (2) only short term recall was tested and, for medical information to be useful clinically, it must be remembered for significant periods of time and (3) a maximum of 50 instructions were shown in pictographs, whereas managing complex illnesses may require remembering several hundred instructions. This study addresses those limitations by investigating 4-week recall of 236 medical instructions accompanied by pictographs by people with low literacy skills. Subjects were 21 adult clients of an inner city job training program who had less than fifth grade reading skills. Results showed 85% mean correct recall of pictograph meanings immediately after training (range from 63 to 99%) and 71% after 4 weeks (range from 33 to 94%). These results indicate that people with low literacy skills can, with the help of pictographs, recall large amounts of medical information for significant periods of time. The impact of pictographs on symptom management and patient quality of life remains to be studied. PMID- 11384822 TI - A comprehensive assessment of satisfaction with care: preliminary psychometric analysis in French, Polish, Swedish and Italian oncology patients. AB - Satisfaction with care may be closely related to quality of life in cancer patients. This evaluation is especially relevant when quality of care is considered. The present study assessed whether equivalent scaling properties could be found in a comprehensive assessment of satisfaction with care (CASC) administered in cancer patients from French, Polish and Swedish oncology settings, in comparison to the scaling properties previously evidenced in the CASC with an Italian sample. A total of 140, 186 and 133 oncology patients were approached in France, Poland and Sweden, respectively. Specific items in the CASC were identified as consistently omitted across country samples. Multitrait scaling analysis on an item-grouping adapted for the French, Polish, Swedish and Italian samples provided excellent internal consistencies and convergent validity estimates. Discriminant validity proved less satisfactory, evidencing overlap between hypothesised care dimensions across country samples. The identification of omitted or overlapping items will lead to the design of a revised CASC version to further test in larger cross-cultural samples. PMID- 11384823 TI - Measuring information needs among cancer patients. AB - A scale for assessing information needs of cancer patients was constructed and validated. Two studies were conducted. Study 1 was designed to test the factor structure of the measurement instrument. A total of 498 patients with breast cancer and Hodgkin disease were interviewed. In study 2, 133 patients with head and neck cancer were measured just before treatment as well as 6, 13 and 52 weeks after treatment. Study 2 aimed to confirm the factor structure established in study 1, and to test for construct validity in a new population, the psychometric properties of the information needs scales, and the scales' sensitivity to change. In study 1 a two-factor structure (an action and a disease-oriented scale) including 17 items was revealed. The second study confirmed the factor structure from study 1. As hypothesized, greater information needs related to higher levels of state-anxiety, more depression, and more psychological complaints. Although, correlations over time per information need scale indicate some stability of scores, findings suggested that the need for information about disease and treatment is less stable over time than need for information about access to help and solutions. Further validation of the instrument is required. PMID- 11384824 TI - Disease knowledge in a high-risk population for cystic fibrosis. AB - Cystic fibrosis (CF) has high incidence (1/936 live births) and carrier rate (1/15 inhabitants) in Saguenay-Lac-Saint-Jean (SLSJ). One objective of a major enquiry among several subsets of individuals from this high-risk population for CF was to evaluate the knowledge of the disease and its genetic transmission. The overall score of correct answers pertaining to the clinical signs of CF among medical doctors (general practitioners and specialists) was 42.2 and 65.6%, respectively; it was 84.2% for questions regarding the genetic transmission of CF. The knowledge of the clinical signs was reasonable among CF patients and their parents (about 65% of correct answers), but it was much higher for the genetics (over 88% among parents). Aunts and uncles of CF children were poorly informed of the clinical signs (33.9% of correct answers) but well informed of the genetic transmission (73.8%). Specific subsets of the SLSJ population showed important gaps in the knowledge of the clinical signs of CF but, overall, they were well informed of its genetic transmission. PMID- 11384825 TI - The short-term impact of tailored mammography decision-making interventions. AB - BACKGROUND: We assessed the short-term impact of decision-making interventions on knowledge about mammography, accuracy of women's breast cancer risk perceptions, attitudes toward mammography, satisfaction with decisions, and mammography use since the intervention. METHODS: The study was conducted among women who were members of Blue Cross Blue Shield of North Carolina and were in their 40s or 50s at the time the study began in 1997. Women were randomly assigned to usual care (UC), tailored print booklets (TP) alone, or TP plus telephone counseling (TP+TC ). RESULTS: 12-month interviews were completed by 1127 women to assess short-term intervention effects. Generally, women who received TP+TC were significantly more knowledgeable about mammography and breast cancer risk and were more accurate in their breast cancer risk perceptions than women in the TP and UC groups. They also were more likely to have had a mammogram since the baseline interview. In multivariable analyses, we found significant benefits of the combination of TP+TC compared to TP and to UC for knowledge, accuracy of risk perceptions, and mammography use. DISCUSSION: For complex decision-making tasks, such as women's decisions about mammography in the face of controversy, the combination of TP and TC may be more effective than TP alone, and certainly more effective than UC. It is critical that investigators determine the topics for which TP is appropriate and the situations that require additional supportive interventions. PMID- 11384826 TI - An educational approach to improving physician breast cancer screening practices and counseling skills. AB - A multifaceted, individualized, physician education program designed to increase the breast cancer screening practices of community-based primary care physicians is described and the results are evaluated. Community-based surveys identified primary care providers with breast cancer screening educational needs who were assigned, using a factorial design, to an intervention or control condition. The sample included 154 control and 128 intervention physicians. The intervention consisted of a 1-2h in-office training program and/or self-study workbook. Self reported overall breast cancer screening need scores improved for a greater proportion of intervention than control physicians, particularly those receiving the in-office intervention (P=0.03). Clinical breast examination (CBE) need declined (P=0.01); use of provider reminder systems increased (P=0.02); preparedness to counsel about CBE (P=0.04) and recognition that age is an important risk factor for breast cancer (P=0.02) improved in more intervention compared to control physicians. PMID- 11384827 TI - Impact of risk information in a patient education leaflet. AB - Informed consent requires the communication of information about possible adverse effects of treatment. Health professionals have expressed concern that increased provision of risk information may make patients unduly anxious and change their decisions about treatment. This study compared two patient leaflets about laparoscopy, one containing detailed information about potential side effects. Forty one consenting participants attending an outpatient gynaecology clinic and scheduled for an elective laparoscopic procedure were studied. They were randomly assigned to receive one of two leaflets after their consultation. Knowledge about laparoscopy, satisfaction with information provided, and anxiety were tested 2 days later, between the original consultation and their operation. Results showed that the leaflet containing detailed risk information was associated with greater knowledge about laparoscopy, higher satisfaction with information provision and no increase in anxiety. Provision of detailed information about possible adverse consequences of treatment can improve patients' understanding and satisfaction without inducing increased anxiety. PMID- 11384828 TI - Managing the psychological aspects of altered appearance: the development of an information resource for people with disfiguring conditions. AB - As the demand for health-related information grows, people are looking beyond health professionals, to patient-led organisations or support groups. The development of an information service is therefore a primary objective for such groups. Why information is needed, what kind of information people want, and who can provide it are central questions. These are discussed in the context of the development of a written information series, produced by a patient-led organisation, for people with facial disfigurement. PMID- 11384829 TI - Time flies! PMID- 11384830 TI - The incidence of glassy corneal striae. AB - Glassy corneal striae (GCS) are fine colorless lines that run parallel to each other in a vertical or near-vertical orientation in the central cornea near the level of Descemet's membrane. It is postulated that GCS "brace" the stroma, thus adding stability to the cornea. The purpose of this study was to determine the incidence of GCS in patients with "normal" corneas and to determine whether a correlation exists between the presence of GCS and gender, age, iris color, or corneal astigmatism (toricity). Modified marginal retro-illumination biomicroscopy was used to evaluate the cornea for the presence of GCS. One hundred thirteen "normal" right corneas were evaluated for the presence of GCS. A total of 85.8% (97/113) of subjects in our study sample exhibited GCS. However, we were unable to establish a statistically significant correlation between the presence of GCS and gender, age, iris color, or corneal toricity. PMID- 11384831 TI - Empirical fitting with polycon II lenses. AB - Historically, rigid gas permeable (RGP) lens parameters have been determined through trial fitting. Nevertheless, practitioners would likely welcome the opportunity to empirically fit RGP lenses if they could be confident of the outcome. This clinical investigation sought to evaluate the potential success of empirically fitting 9.0 mm Polycon II lenses. Based on both objective and subjective outcomes, this study suggests that empirically fitting the larger 9.5 mm Polycon II design, in combination with a lid attachment approach, would yield greater initial patient satisfaction. PMID- 11384832 TI - Discontinuation of contact lens wear: a survey. AB - Discontinuation from contact lens wear has been cited as a contributing factor to the lack of growth of the contact lens market in North America. To determine the reasons and extent of abandonment from contact lens wear among patients from optometric practices, 4415 questionnaires were randomly mailed to patients from 16 practices in Quebec, Canada, and were returned as anonymous responses to the Centre for Contact Lens Research in Waterloo. Thirty-three percent (n = 1444) of the surveys were returned. After 5 +/- 6 years of lens wear, 34% of patients had discontinued lens wear at least once. Of the patients who discontinued, 77% resumed wear and 48% of those discontinued a second time. Those who had discontinued were fitted at an older age (by approximately 2 years), were in the professional/health care/management occupation sector, and had a slightly lower myopic prescription. Before discontinuation, the wearing time of this group was approximately 1 day/week less than those continuing to wear lenses. Of the patients who had discontinued, 49% had been refitted at least once, and the primary reason for refitting was discomfort. The primary reasons for abandonment were discomfort, dryness, and red eyes. Improved comfort and relief of dryness were the primary factors that would improve contact lens wear for the group that had discontinued. Practitioners may be able to target specific patient groups with remedial measures to improve contact lens-wearing success. PMID- 11384833 TI - Complexity of contact lens fitting following penetrating keratoplasty. AB - The aim of this study was to quantify the complexity of contact lens (CL) fitting following penetrating keratoplasty (PKP).The CL care of 40 eyes following PKP was analyzed retrospectively and compared to that of 40 age-matched and gender matched controls. We evaluated the numbers of diagnostic and ordered rigid gas permeable (RGP) CLs, office visits for 6-month follow-up, best spectacle and RGP CL-corrected logMar visual acuities (VAs), and RGP CL success and complication rates. Post-PKP corneas required more diagnostic CLs (p = 0.009), ordered CLs (p = 0.0003), and office visits (p = 0.001) than did controls. Corrected logMAR VAs post-PKP improved from 0.31 +/- 0.31 (mean +/- SD) with spectacles (20/41) to 0.076 +/- 0.19 (20/24) with RGP CLs (p < 0.0001). Controls logMAR spectacle and CL-corrected VAs were 0.0043 +/- 0.12 (20/20) and -0.023 +/- 0.058 (20/19), respectively (no significant difference, p = 0.096). Ninety percent of PKP eyes and 100% of controls were successful, and 30% of PKP and 12.5% controls had CL related complications; these differences did not reach statistical significance.Post-PKP eyes require more diagnostic CLs, ordered CLs, and professional office visits. They also have greater improvement of VAs with RGPs than normals compared to spectacles. CL wear post-PKP does not statistically increase the rate of CL complications or alter the success rate of CL wear. PMID- 11384834 TI - Accuracy and reproducibility of one-day disposable contact lenses. AB - One-day disposable contact lenses are experiencing rapid market growth in certain markets, such as the United Kingdom. Because of the huge numbers of lenses required to satisfy this modality of wear, both practitioners and patients need to be reassured about the quality of these products. Analysis of the accuracy and reproducibility of each of three currently available 1-day disposable lenses was conducted with respect to the following lens parameters: total diameter, back optic zone radius, center thickness, water content, and back vertex power. Fourteen of the 15 parameters measured (three lenses x five parameters per lens) were found to fall within ISO-defined tolerances. Reproducibility was similar for the three lens types and was considered clinically acceptable. Practitioners and patients can be confident that the current generation of 1-day disposable lenses can be relied on to give consistent ocular and visual performance. PMID- 11384835 TI - Four views of hypoxiaellipsis, but which is best? part 2. using air as an ideal reference. PMID- 11384836 TI - Phosphoinositides: key players in cell signalling, in time and space. AB - Over the last few years, many reports have extended our knowledge of the inositol lipid metabolism and brought out some exciting information about the location, the variety and the role of phosphoinositides (PIs). Besides the so-called "canonical PI pathway" leading to the production of phosphatidylinositol 4,5 bisphosphate (PtdIns(4,5)P2), the precursor of the intracellular second messengers inositol 1,4,5-trisphosphate and diacylglycerol (DAG), many other metabolic pathways have been identified to produce seven different polyphosphoinositides. Several of these quantitatively minor lipid molecules appear to be specifically involved in the control of cellular events, such as the spatial and temporal organisation of key signalling pathways, the rearrangement of the actin cytoskeleton or the intracellular vesicle trafficking. This is consistent with the fact that many of the enzymes, such as kinases and phosphatases, involved in the tight control of the intracellular level of polyphosphoinositides, are regulated and/or relocated through cell surface receptors for extracellular ligands. The remarkable feature of PIs, which can be rapidly synthesised and degraded in discrete membrane domains or even subnuclear structures, places them as ideal regulators and integrators of very dynamic mechanisms of cell regulation. In this review, we will summarise recent studies on the potential location, the metabolic pathways and the role of the different PIs. Some aspects of the temporal synthesis of D3 PIs will also be discussed. PMID- 11384837 TI - The TNF-receptor-associated factor family: scaffold molecules for cytokine receptors, kinases and their regulators. AB - The TNF-receptor-associated factor (TRAF) family is a phylogenetically conserved group of scaffold proteins that link receptors of the IL-1R/Toll and TNF receptor family to signalling cascades, leading to the activation of NF-kappaB and mitogen activated protein kinases. Furthermore, TRAF proteins serve as a docking platform for a variety of regulators of these signalling pathways and are themselves often regulated at the transcriptional and posttranslational level. In this review, we address the structural and molecular basis of TRAF protein functions and highlight their role in cytokine signalling. PMID- 11384838 TI - Platelet-derived growth factor mediated altered expression and regulation of ornithine decarboxylase in H-ras-transformed cell lines. AB - This study demonstrates a novel link between alterations in platelet-derived growth factor (PDGF) regulation of ornithine decarboxylase (ODC) expression during malignant conversion. H-ras-transformed cell lines exhibited PDGF-mediated alterations in ODC gene expression. These alterations involved transcriptional, posttranscriptional, and cycloheximide-mediated events. PDGF-mediated alterations in ODC gene expression in NR3 cells (capable of only benign tumour formation) were ras-dependent, involved a tyrosine kinase activity and mitogen-activated protein (MAP) kinase-mediated signalling events, and were independent of both protein kinase C (PKC) events and pertussis toxin-sensitive (PTS) G-protein mediated signalling. PDGF-mediated alterations in ODC gene expression in C2 cells [capable of malignant progression (metastasis formation)] were ras-dependent, required a tyrosine kinase activity, involved both MAP kinase-mediated events and phosphatidylinositol-3-kinase (PI-3-kinase)-mediated events, and were dependent upon PTS G-protein-mediated signalling but independent of PKC-mediated events. PDGF-mediated regulation of ODC gene expression changes in response to H-ras mediated cellular transformation and malignant progression. PMID- 11384839 TI - SH3 ligands in the dopamine D3 receptor. AB - It has recently been observed that G protein-coupled receptors (GPCRs) can interact with SH3 domains through polyproline motifs. These interactions appear to be involved in receptor internalization and MAPK signalling. Here we report that the third cytoplasmic loop of the dopamine D3 receptor can interact in vitro with the adaptor protein Grb2. While the amino- and carboxy-terminal SH3 domains of Grb2 separately did not interact with the D3 receptor loop, the interaction is at least partially maintained with a Grb2 mutant for the amino-terminal SH3 domain, but disrupted for a Grb2 mutant with a nonfunctional carboxy-terminal SH3 domain. The data indicate the need of structural integrity of the entire Grb2 protein for the interaction and dominant role of the carboxy-terminal SH3 domain in the interaction. Disruption of the PXXP motifs in the D3 receptor did not affect the interaction with Grb2. These results indicate that GPCRs may contain SH3 ligands that do not contain the postulated minimal consensus sequence PXXP. PMID- 11384840 TI - Activation of c-Jun N-terminal kinase 1 (JNK-1) after amino acid deficiency in HeLa cells. AB - Long-term amino acid starvation represents a form of metabolic stress which stimulates gene expression. Here we report that depriving HeLa cells for any one of a series of amino acids activates c-Jun N-terminal kinase-1 (JNK-1). In contrast, the other mitogen-activated protein kinases (MAPKs) ERK-1 and, to a lesser extent, p38 activities decreased under such conditions. In methionine- or leucine-deprived cells, JNK-1 activation occurred after 4 or 6 h, respectively, and reached a steady maximum of 5- to 7-fold over control cells afterwards. This activation was dependent on the amino acid concentration and it could be reversed by resupplying the complete medium. Limitation for all amino acids also augmented JNK-1 activity, whereas increased amino acid concentrations had an opposite effect. The free radical scavenging thiol antioxidant N-acetylcysteine (NAC) alleviated partially JNK-1 activation in amino acid-deprived cells. The data indicate that activation of JNK-1 by long-term amino acid deprivation may be mediated in part by oxidative stress. PMID- 11384841 TI - Cyclic GMP-dependent protein kinase potentiates serotonin-induced Egr-1 binding activity in PC12 cells. AB - The NO/cyclic GMP (cGMP) signal transduction pathway, which involves the cGMP dependent protein kinase (PKG), regulates transcription of several genes, including immediate early genes. Using transfection experiments with the PKG Ialpha cDNA cloned from human aorta, we show here that addition of membrane permeable cGMP analogues to PC12 cells slightly upregulated ERK MAP (mitogen activated protein) kinase. Likewise, PKG-Ialpha was found to activate weakly DNA binding activity of the Egr-1 transcription factor. On the other hand, PKG-Ialpha overexpression was shown to tremendously amplify the Egr-1 binding activity induced by the neurotransmitter serotonin, which activates egr-1 gene expression also via the stimulation of the ERK MAP kinase pathway. Since this potentiation occurred neither at the level of ERK nor at the egr-1 transcriptional level, the mechanism of amplification probably results from the convergence of ERK and PKG pathways at the level of the transcription factor Egr-1. PMID- 11384842 TI - Protein kinase C isozyme-specific phosphorylation of profilin. AB - Profilin, a cytoskeletal protein, is emerging as an important link between signal transduction pathways and cytoskeletal dynamics. Profilin is phosphorylated on its C-terminal serine by protein kinase C (PKC). The protein kinase used for the in vitro phosphorylation studies reported earlier was a mixture of isozymes, and therefore, attempts were made to address the isozyme specificity on profilin phosphorylation under in vitro conditions. Profilin was subjected to phosphorylation by PKCalpha, PKCepsilon, and PKCzeta isozymes individually, and it was observed that profilin phosphorylation is cofactor-independent. PKCzeta phosphorylates profilin to a higher extent, but exhibits cofactor dependency with respect to phosphoinositides. The stoichiometry of phosphorylation was measured in the presence of these different isozymes, and a maximum stoichiometry of 0.8 (mole phosphate incorporated/mole profilin) was obtained in the presence of PKCzeta. Phosphorylation of profilin by PKCzeta was maximal in the presence of phosphatidylinositol4,5-bisphosphate (PI4,5-P2) when compared to the other phosphoinositides studied. PMID- 11384843 TI - Modulation of thapsigargin-induced calcium mobilisation by cyclic AMP-elevating agents in human lymphocytes is insensitive to the action of the protein kinase A inhibitor H-89. AB - Ca2+ mobilisation from internal stores and from the extracellular medium is one of the primary events involved in lymphocyte activation and proliferation. Regulation of these processes by adenosine 3',5'-cyclic monophosphate (cAMP) and cAMP-dependent protein kinase (PKA) was studied in Fura2-loaded human peripheral blood lymphocytes. Cytosolic Ca2+ concentration ([Ca2+]i) was measured in single cells by the use of a ratio imaging fluorescence microscope and Ca2+ mobilisation was achieved by the use of the endoplasmic reticulum (ER) Ca2+ ATPase inhibitor, thapsigargin (Thg). Our results show that both activation and inhibition of PKA, with forskolin (FSK) and N-[2-(p-bromocinnamylamino)ethyl]-5 isoquinolinesulfonamide.2HCl (H-89), respectively, inhibited the Thg-induced Ca2+ entry. Furthermore, FSK also reduced the ability of Thg to release Ca2+ from internal stores. This reduction was inhibited by the adenylyl cyclase (AC) inhibitor 9-(tetrahydro-2-furanyl)-9-H-purin-6-amine (SQ22,536), but not by the PKA inhibitor H89, indicating that cAMP but not PKA is responsible for this effect. FSK effect was mimicked by dibutyryl cAMP (dbcAMP) and by inhibition of phosphodiesterases (PDEs) with rolipram (ROL) and milrinone (MIL). We also showed that a very high concentration of H-89 (100 microM) releases Ca2+ from an intracellular pool, although this action is probably independent of PKA inhibition. Neither 10 microM H-89 nor other cAMP/PKA-modulating drugs had any effect on the basal [Ca2+]i of human lymphocytes. We conclude that PKA may act as a fine modulator of capacitative Ca2+ entry, while cAMP has a PKA-independent interaction with the Ca2+ stores of human lymphocytes. PMID- 11384844 TI - Vanadate inhibits the calcium extrusion in rat pancreatic acinar cells. AB - Our objective was to evaluate the role of vanadate on calcium extrusion in Fura-2 loaded rat pancreatic acinar cells by digital microscopic fluorimetry and spectrofluorimetry. In the absence of extracellular calcium, perfusion of pancreatic acinar cells with 1 nM CCK-8 and 1 mM vanadate did not significantly affect the typical transient calcium spike induced by CCK-8, but the plateau phase of calcium in response to CCK-8 remained elevated. In addition, vanadate was able to inhibit calcium efflux evoked by CCK-8 when we determined directly calcium transport across plasma membrane using Calcium Green-5N hexapotassium salt (cell impermeant form) in cell populations. The effect of vanadate on calcium extrusion was strongly blocked by the sulfhydryl-reducing agent dithiothreitol (DTT). The present results demonstrate that vanadate is able to irreversibly inhibit the calcium extrusion. This effect of vanadate can be blocked using DTT, indicating that its action is probably mediated by oxidation of sulfhydryl groups of Ca2+-ATPases. PMID- 11384846 TI - In vitro evaluation of 2,4-dihydroxythiobenzanilides against various moulds. AB - The antimycotic potency of 2,4-dihydroxythiobenzanilide derivatives was tested. The MIC assessments by an agar dilution method were used for the estimation of potential activity in vitro against the four mould strains: Scopulariopsis brevicalis, Aspergillus niger, Aspergillus fumigatus and Penicillium sp. The strongest fungistatic activity was observed for 3'-fluoro-derivative (MIC 7.82 microg/ml). It was stated that the inhibition action of these compounds depends mainly on lipophilicity of molecules. Parabolic relationships between the antimycotic activity and lipophilicity were found. PMID- 11384847 TI - Influence of imidazole replacement in different structural classes of histamine H(3)-receptor antagonists. AB - The reference compounds for histamine H(3)-receptor antagonists carry as a common feature an imidazole moiety substituted in the 4-position. Very recently novel ligands lacking an imidazole ring have been described possessing a N-containing non-aromatic heterocycle instead. In this study we investigated whether imidazole replacement, favourably by a piperidine moiety, is generally applicable to different structural classes of reference compounds, e.g., thioperamide, carboperamide, clobenpropit, FUB 181, ciproxifan, etc. While replacement led to a loss of affinity for many of the compounds, it was successfully applied to some ether derivatives. The piperidine analogues of FUB 181 and ciproxifan, 3-(4 chlorophenyl)propyl 3-piperidinopropyl ether hydrogen oxalate (6) and cyclopropyl 4-(3-piperidinopropyloxy)phenyl methanone hydrogen maleate (7), almost maintained in vitro affinities, pK(i) values of 7.8 and 8.4, respectively, and showed high potency in vivo after p.o. administration (ED(50) values of 1.6 and 0.18 mg/kg, respectively). PMID- 11384848 TI - Computer-assisted drug development (CADD): an emerging technology for designing first-time-in-man and proof-of-concept studies from preclinical experiments. AB - Computer-assisted drug development (CADD) is an emerging technology for accelerating drug development based on the integration of mathematical modelling and simulation. This methodology provides a knowledge-based decisional tool on alternative development strategies based on the evaluation of potential risks on drug safety, and the definition of experimental design of new trials with expected power and probability of success. An example of CADD implementation is presented to design the first-time-in-man (FTIM) and the proof-of-concept (PoC) study of a new CNS compound. The final objective of the example presented is not necessarily to supply a success story of a correct prediction of human data from animal studies but to define a credible strategy suitable to design FTIM and PoC studies using preclinical data without the support of any human in vivo information. Rhesus monkey and human PK were initially estimated using allometric scaling on data collected in dogs, cynomolgus monkeys and rats. A PK/PD model was derived from a study conducted in rodent and validated by comparing the model predicted response to the one observed in a PET experiment conducted in rhesus monkey. The final PK/PD model, incorporating potential variability and uncertainty on scaled human prediction together with a receptor affinity adjustment derived from in vitro binding studies, was used to design the first time-in-man and the proof-of-concept study. PMID- 11384849 TI - Molecular modeling study of beta- and gamma-cyclodextrin complexes with miconazole. AB - Different authors have demonstrated the inclusion of miconazole in cyclodextrins (CD). Miconazole can be included in the CD cavity both in the neutral and in the ionized form. The present study tries to understand which fragment of the miconazole molecule is involved in the inclusion. Austin Model 1 approximate molecular orbital calculations have been performed on several complexes between beta-cyclodextrin (betaCD) or gamma-cyclodextrin (gammaCD) and miconazole in the ionized and the non-ionized forms of the two R and S enantiomers in three different orientations. We observed that betaCD is a good vehicle to transport miconazole which can be very easily released. The complexation energy between miconazole and betaCD is not very high but the entropic factor has a great incidence on the stability of the formed complex. The inclusion of the dichlorobenzene-CH(2)-O- and of the imidazole part of the S isomer gives rise to the most probable complex in acidic conditions (ionized miconazole). Nevertheless, the inclusion should be considered as a dynamic process in which different parts of the molecule could be alternatively included in betaCD. The present work demonstrates the high capability of deformation of betaCD which could easily accommodate several types of ligand. By opposite, the cycle extension in gammaCD leads to a more rigid vehicle with regards to miconazole. PMID- 11384850 TI - Solubility and conversion of carbamazepine polymorphs in supercritical carbon dioxide. AB - The aim of this work was to investigate whether mixtures of carbamazepine polymorphs could be processed in supercritical (SC) CO(2) in order to obtain the pure stable crystalline phase. To accomplish this goal the solubility of carbamazepine polymorphs I and III in supercritical CO(2) was first assessed using a low solvent flux dynamic method. Mixtures of Form I and Form III were processed in dynamic or static conditions in SC-CO(2). Differential scanning calorimetry, Fourier transformed infrared spectroscopy, and powder X-ray diffractometry were used to analyse solid samples in terms of polymorph composition. It was found that Form I and Form III of carbamazepine have different solubility in supercritical CO(2) at 55 degrees C above 300 bar. Due to the transformation of the metastable form, conversion of Form I into Form III can be carried out on a binary mixture of the two polymorphs by treating the mixture at 55 degrees C and 350 bar, under both static and dynamic conditions, via its solubilization in supercritical CO(2). PMID- 11384851 TI - Liposomes encapsulating prednisolone and prednisolone-cyclodextrin complexes: comparison of membrane integrity and drug release. AB - Inclusion complexes of prednisolone (PR) with beta-cyclodextrin (beta-CD) and hydropropyl-beta-cyclodextrin (HPbeta-CD) were formed by the solvation method, and were characterized by DSC, X-ray diffractometry and FT-IR spectroscopy. PC liposomes incorporating PR as plain drug or inclusion complex were prepared using the dehydration-rehydration method and drug entrapment as well as drug release were estimated for all liposome types prepared. The highest PR entrapment value (80% of the starting material) was achieved for PC/Chol liposomes when the HPbeta CD-PR (2:1, mol/mol) complex was entrapped. The leakage of vesicle encapsulated 5,6-carboxyfluorescein (CF) was used as a measure of the vesicle membrane integrity. As judged from our experimental results liposomes which encapsulate beta-CD-PR complexes are significantly less stable (when their membrane integrity is considered) compared to liposomes of identical lipid compositions which incorporate plain drug or even (in some cases) non-drug incorporating liposomes, which were prepared and studied for comparison. Interestingly, liposomes which encapsulate HPbeta-CD-PR complexes, have very low initial CF latency values, indicating that the leakage of CF is a process of very high initial velocity. Interactions between lipid and cyclodextrin molecules may be possibly resulting in rapid reorganization of the lipid membrane with simultaneous fast release of CF molecules. The release of PR from liposomes was highest when the drug was entrapped in the form of a complex with beta-CD. Nevertheless, the very high entrapment ability of PR in the form of HPbeta-CD-PR complexes in comparison to plain drug is a indubitable advantage of this approach. PMID- 11384852 TI - Determination of trovafloxacin in human serum by time resolved terbium-sensitised luminescence. AB - A sensitive time-resolved luminescence method for the determination of trovafloxacin is described. The method is based on the time-resolved luminescence signal from the terbium(III)-trovafloxacin complex, in a micellar solution of sodium dodecyl sulfate (SDS), using a chemical deoxygenation agent (Na(2)SO(3)). The method allows the determination of 20-450 ng ml(-1) of trovafloxacin in 7.5 mM SDS solution containing 0.16 M acetic acid-sodium acetate buffer (pH 6.0) and 7.5 mM Na(2)SO(3) with lambda(exc)=270 nm and lambda(em)=546 nm. In these experimental conditions luminescence signal for trovafloxacin increases 20-fold with respect to native fluorescence of the compound in aqueous solution at pH 6.5. Terbium-sensitised luminescence was applied to trovafloxacin determination in human serum, spiked at levels found after drug administration at normal clinical doses. Recovery is 90+/-1% and day-to-day precision is 3.5%. The proposed method tolerates high concentrations of other co-administrated drugs and excipients. PMID- 11384853 TI - Quantitative estimation of renal clearance of N-acetylprocainamide in rats with various experimental acute renal failure. AB - The dosage regimen of a drug eliminated predominantly through the kidney need to be adjusted for the patients with renal disease. The objective of the present study was to establish a quantitative approach to precisely predicting the renal clearances of basic drugs using N-1-methylnicotinamide (NMN). A variety of experimental acute renal failure (ARF) in rats were prepared and N acetylprocainamide (NAPA) was used as a model drug. The renal clearances of NAPA were significantly decreased in rats with ARF, resulting in significantly increased plasma concentrations. Remarkable reduction in clearance ratios (CL(ratio)) was observed, indicating that the impairment in tubular and glomerular function did not proceed in a parallel manner. The renal clearance of NAPA (CL(rNAPA)) was better predicted from the renal clearance of NMN (CL(rNMN)) than from GFR. A mathematical equation was also constructed to estimate the CL(rNMN) from the NMN plasma concentration. Therefore, the renal clearance of basic drugs excreted predominantly from the kidney can be easily and more accurately estimated based on the concentrations of endogenous NMN to provide a precise dosage regimen for patients with renal failure. PMID- 11384854 TI - The effect of rheological behaviour of a topical anaesthetic formulation on the release and permeation rates of the active compound. AB - The objective of this study was to investigate the possibility of developing a topical cream that allows maximum release rate of the active compound while having suitable consistency, i.e., sufficient apparent plasticity. A submicron (o/w) emulsion containing a model compound was investigated in the presence and absence of different polymers: sodium carboxymethylcellulose (CMC), Carbopol 934P (C934), polyethylene glycol 400 (PEG400) and polyethylene glycol 4000 (PEG4000). Various concentrations of the polymers were used in order to produce different rheological behaviours. The amount of drug passing through the membrane was measured as a function of time, using static diffusion cells with either Silastic sheeting 500-1 or guinea pig skin as membrane. The emulsion without polymer was used as reference. Rheological measurements were performed, giving the viscosity and the apparent yield stress of the formulations. Furthermore, theoretical values for diffusion coefficients and diffusion pathways were estimated and compared with the experimental data to discuss different diffusion models. Gelling polymers have been shown to produce an increase in the macroviscosity, thus inhibiting the diffusion of the oil droplets in the formulation without affecting the molecular diffusion. However, we suggest that when a compound of limited solubility is emulsified, the intact oil droplets contribute to the transport of the compound through the formulation. Thus, both release and permeation rates are decreased as the apparent yield stress, i.e., the macroviscosity of the formulation, is increased sufficiently by addition of gelling polymers. PMID- 11384855 TI - Endotracheal tube cuffs filled with lidocaine as a drug delivery system: in vitro and in vivo investigations. AB - The purpose of this study was to examine if lidocaine diffusion across an endotracheal tube cuff could improve post-operative tolerance, especially sore throat. The in vitro release of lidocaine from tube cuffs filled with different lidocaine formulations (base form, hydrochloride form or alkalinized lidocaine hydrochloride) was investigated. A preliminary pilot clinical study in anaesthesia for spine surgery in smoker patients was carried out to examine the pharmacokinetic (i.e. systemic uptake) and pharmacodynamic effects (i.e. incidence of sore throat) obtained with the endotracheal tube cuff filled with lidocaine solution, compared to cuffs inflated only with air. From our in vitro experiment, only the hydrophobic neutral base form of lidocaine was able to diffuse (65.1+/-1.1% released after 6 h), while for the charged hydrochloride form, only a permeation phenomenon occurred concerning only 1% of the total drug. Alkalinization of lidocaine hydrochloride (the only form available as a drug) allows smaller amounts to be used compared to previous published studies (20-40 mg vs. 200-500 mg) and no lag time for diffusion. Such a system could provide a controlled release reservoir for lidocaine to adjacent tracheal tissue. This was shown in our pilot study with sustained plasmatic profiles and improved tolerance (decreased pain scores) in the rank order: air group<51 years). This suggests that VD has a protective role against prostate cancer only before the andropause, when serum androgen concentrations are higher. The lowest 25-VD concentrations in the younger men were associated with more aggressive prostate cancer. Furthermore, the high 25-VD levels delayed the appearance of clinically verified prostate cancer by 1.8 years. Since these results suggest that vitamin D has a protective role against prostate cancer, we tried to determine whether full spectrum lighting (FSL) during working hours could increase serum 25-VD concentrations. After 1-month exposure, there was no significant increase in the serum 25-VD level, although there was a bias towards slightly increasing values in the test group as opposed to decreasing values in controls. There was no significant change in the skin urocanic acid production. The possibility to use FSL in cancer prevention is discussed. In order to clarify the mechanism of VD action on cell proliferation and differentiation, we performed studies with the rat and human prostates as well prostate cancer cell lines. It is possible that 25-VD may have a direct role in the host anticancer defence activity, but the metabolism of vitamin D in the prostate may also play an important role in its action. We raised antibodies against human 1alpha hydroxylase and 24-hydroxylase. Our preliminary results suggest that vitamin D is actively metabolised in the prostate. Vitamin D appears to upregulate androgen receptor expression, whereas androgens seem to upregulate vitamin D receptor (VDR). This may at least partially explain the androgen dependence of VD action. VD alone or administered with androgen causes a suppression of epithelial cell proliferation. VD can activate mitogen-activated kinases, erk-1 and erk-2, within minutes and p38 within hours. Also, auto/paracrine regulation might be involved, since keratinocyte growth factor (mRNA and protein) was clearly induced by VD. Based on these studies, a putative model for VD action on cell proliferation and differentiation is presented. PMID- 11384871 TI - The androgen receptor (AR) in syndromes of androgen insensitivity and in prostate cancer. AB - The actions of androgens, principally testosterone and 5alpha dihydrotestosterone, are mediated by a specific receptor protein, the androgen receptor (AR), which is encoded by a single-copy gene located on the human X chromosome. This receptor protein is a prototypical member of the nuclear receptor family and modulates a range of processes during embryogenesis and in the adult. During embryogenesis, normal AR function is critical to the development of the male phenotype and defects of the AR cause a range of phenotypic abnormalities of male sexual development. Complete loss of AR function has been traced to a number of distinct types of genetic events, including abnormalities of mRNA splicing, the introduction of premature termination codons, and amino acid substitution mutations. An interesting subset of mutations is that in which the AR is completely undetectable using sensitive immunoassays. In all instances, these functional abnormalities are associated with a phenotype of complete androgen insensitivity (complete testicular feminization). By contrast, partial defects of AR function are almost invariably caused by amino acid substitutions within the DNA- and hormone-binding domains of the receptor protein. Such partial defects of receptor function may be caused by changes in either receptor function or receptor abundance. The alterations of AR function and expression that have been characterized in clinical prostatic cancers and in prostate cancer cell lines differ in several important respects. A number of studies have documented the emergence of considerable heterogeneity of AR expression at early stages in the development of prostate cancer. Despite these early changes of AR expression, a substantial body of information suggests that the AR is expressed in advanced forms of prostate cancer, in some cases as the result of amplification events. While infrequent in localized tumors, mutations of the AR have been identified in a number of advanced prostatic cancers and in some instances appear to alter the ligand specificity of the AR. Finally, it appears that other signaling pathways can act to influence AR function. PMID- 11384872 TI - Analysis and characteristics of multiple types of human 17beta-hydroxysteroid dehydrogenase. AB - Androgens and estrogens are not only synthesized in the gonads but also in peripheral target tissues. Accordingly, recent molecular cloning has allowed us to identify multiple types of 17beta-hydroxysteroid dehydrogenases (17beta-HSD), the key and exclusive enzymes involved in the formation and inactivation of sex steroids. However, only one form, namely, type 3 17beta-HSD, is responsible for pseudohermaphroditism in deficient boys. To date, seven human 17beta-HSDs have been isolated and characterized. Although they catalyze substrates having a similar structure, 17beta-HSDs have very low homology. In intact cells in culture, these enzymes catalyze the reaction in a unidirectional way - types 1, 3, 5 and 7 catalyze the reductive reaction, while types 2, 4 and 8 catalyze the oxidative reaction. It is noteworthy that rat type 6 17beta-HSD also catalyzes the reaction in the oxidative direction. In this report, we analyze the different characteristics of the multiple types of human 17beta-HSD. PMID- 11384873 TI - Expression of the 11beta-hydroxysteroid dehydrogenase type II enzyme in breast tumors and modulation of activity and cell growth in PMC42 cells. AB - Manipulating the metabolism of glucocorticoids may serve as a useful adjunct in the treatment of breast cancer. The 11beta-hydroxysteroid dehydrogenase type 2 enzyme (11betaHSD2) potently inactivates glucocorticoids thereby protecting the non-selective mineralocorticoid receptor (MR) in fluid transporting tissues. In the present study, Western blot analysis showed the presence of 11betaHSD2 in 66% of the breast tumor samples. The 11betaHSD2 and MR are also present in the breast tumor cell line PMC42. Glycyrrhetinic acid abolished glucocorticoid metabolism and inhibited cell growth by 40%, the latter at concentrations consistent with glucocorticoid receptor (GR) and MR binding studies. Metabolism was increased by glucocorticoids, the anti-glucocorticoid RU 38486 and anti-mineralocorticoid spironolactone, while aldosterone had no effect. Neither cortisol nor aldosterone affected cell proliferation, but both RU 38486 and spironolactone caused a significant decrease in cell number. The effects of RU 38486 were only observed at micromolar concentrations and are inconsistent with an action via GR or progesterone receptor (PR). This study shows that 11betaHSD2 activity and cell proliferation of PMC42 cells can be modulated via steroid receptors. PMID- 11384874 TI - Resistance to multiple steroids in two sisters. AB - A 14-year-old Native American girl from the Iroquois Nation was referred as a potential patient with the syndrome of Apparent Mineralocorticoid Excess. Instead, her evaluation revealed resistance to glucocorticoids, mineralocorticoids, and androgens. She lacked Cushingoid features in spite of significantly high cortisol levels. Menstruation was regular and there was no clinical evidence of masculinization despite high serum androgen levels in the male range. The patient's sister had similar clinical features. Partial resistance to exogenous glucocorticoid and mineralocorticoid administration was well demonstrated in both patients. It is proposed that these patients represent the first cases of partial resistance to multiple steroids, possibly owing to a coactivator defect. PMID- 11384875 TI - Interaction of thyroid hormone and steroidogenic acute regulatory (StAR) protein in the regulation of murine Leydig cell steroidogenesis. AB - The steroidogenic acute regulatory (StAR) protein, a novel phosphoprotein, is a crucial factor involved in intramitochondrial cholesterol transportation, the rate-limiting step in steroidogenesis. The present investigations were undertaken to elucidate involvement of thyroid hormone and StAR protein in the regulation of steroidogenesis in mouse Leydig cells. Treatment of cells with triiodothyronine (T(3)) coordinately augmented the levels of StAR protein, StAR mRNA, and steroid production, and these responses were progressively dependent on expression of steroidogenic factor 1 (SF-1). With regard to steroidogenesis and StAR expression, the T(3) response requires both on-going mRNA and protein synthesis. In addition, the effects of T(3) were acutely modulated at the steroidogenic machinery and luteinizing hormone receptor (LHR) function, while these levels were suppressed following longer periods of exposure to T(3). Furthermore, the inhibition of SF-1 expression by DAX-1 markedly abolished T(3)-mediated StAR expression in a time frame, which was consistent with decreased steroid biosynthesis. Specific involvement of SF-1 was further confirmed by assessing the 5'-flanking region of the mouse StAR gene, which identified a region between -254 and -110 bp that was essential for T(3) function. Importantly, it was found that the SF-1 binding site at position -135 bp of the 5'-flanking region was greatly involved in T(3)-mediated reporter activity. Electrophoretic mobility shift assays (EMSA) also demonstrated involvement of SF-1 in T(3) function. The relevance of T(3)-mediated LHR function was investigated in mice rendered hypo and hyperthyroid, which accounted for up-regulation in the former and down regulation in the latter group, respectively. These findings demonstrate a key role of thyroid hormone in maintaining mouse Leydig cell function, where thyroid hormone and StAR protein coordinately regulate steroid hormone biosynthesis. PMID- 11384876 TI - Recent advances in the clinical application of aromatase inhibitors. AB - Aromatase inhibitors have evolved over a period of 20 years to well tolerated agents that can effectively obliterate aromatase activity in postmenopausal women. Breast cancer is the predominant clinical application and here the newer agents have established themselves as the preferred second-line agent after tamoxifen in the treatment of advanced disease. Recent data indicate that they be more efficacious than tamoxifen and, therefore, may replace it as the first-line agent of choice in the near future. On-going clinical trials in the adjuvant setting and prospective prevention studies will elucidate whether these drugs have a yet greater role in breast cancer. PMID- 11384877 TI - Regulation of steroidogenic enzymes and a novel testicular RNA helicase. AB - Luteinizing hormone (LH) supports steroidogenesis and maintains testicular and ovarian function. Mediators of LH action exert homologous regulation of membrane receptors, steroidogenic enzymes and other regulatable genes of the Leydig cell (LC). Androgen and estrogen induced by LH could act through its cognate receptors in the LC to regulate gene expression. Although androgens are unquestionable essential for spermatogenesis and presumably exert their heterologous action through androgen receptors present in the Sertoli its regulatory mechanism in germinal cell maturation is far from clear. In contrast to physiological concentrations of gonadotropins which maintain the steroidogenic functions and LH and prolactin receptors in the gonads, high concentrations of gonadotropin (hCG) cause receptor down-regulation and desensitization of steroidogenic enzymes of the LCs in vivo (3beta-hydroxysteroid dehydrogenase types I and II, 17alpha hydroxylase/17,20 lyase, and 17beta-hydroxysteroid dehydrogenase type III [17beta HSD]). In addition, 17beta-HSD is regulated by compartmentalized endogenous glucose/ATP. The attenuation of steroidogenesis which results from receptor mediated activation by cognate hormone, but is independent of the subsequent phase of receptor down-regulation, is due to changes at the transcriptional level. Among the candidates affecting this regulation are active steroid metabolites (direct or indirect of steroids and other mediator(s) i.e. cAMP, putative transcription factors induced by LH action). Differential display assay revealed another gene which is transcriptionally regulated by gonadotropin termed GRTH (Gonadotropin Regulated Testicular Helicase). GRTH is a novel member of the DEAD-box family of RNA helicases, and is specifically expressed in LCs and meiotic LC of the testis. It is markedly up-regulated by hCG via cAMP-induced androgen formation in LCs at doses that cause down-regulation of receptors and steroidogenic enzymes. GRTH functions as a translational activator. Androgen produced by gonadotropin stimulation exerts intracrine/autocrine actions on GRTH, and also could influence transcription within the seminiferous tubule. GRTH may contribute to the control of steroidogenesis, including the restoration of down regulated cellular functions, and in the paracrine regulation of androgen dependent gene(s) involved in the meiotic process, and could thus have a crucial role in spermatogenesis. PMID- 11384878 TI - Intracellular aromatase and its relevance to the pharmacological efficacy of aromatase inhibitors. AB - An important feature of the pharmacological profile of aromatase inhibitors is the ability of the various inhibitors to inhibit intracellular aromatase. It is now well documented that a large proportion of breast tumors express their own aromatase. This intratumoral aromatase produces estrogen in situ and therefore may contribute significantly to the amount of estrogen to which the cell is exposed. Thus it is not only important that aromatase inhibitors potently inhibit the peripheral production of estrogen and eliminate the external supply of estrogen to the tumor cell, but that they in addition potently inhibit intratumoral aromatase and prevent the tumor cell from making its own estrogen within the cell. To study the inhibition of intracellular aromatase we have compared the aromatase-inhibiting potency of the non-steroidal aromatase inhibitors, letrozole, anastrozole and fadrozole in a variety of model cellular endocrine and tumor systems which contain aromatase. We have used hamsters ovarian tissue fragments, adipose tissue fibroblasts from normal human breast, the MCF-7Ca human breast cancer cell line transfected with the human aromatase gene and the JEG-3 human choriocarcinoma cell line. Although letrozole and anastrozole are approximately equipotent in a cell-free aromatase system (human placental microsomes), letrozole is consistently 10-30 times more potent than anastrozole in inhibiting intracellular aromatase in intact rodent cells, normal human adipose fibroblasts and human cancer cell lines. Whether these differences between letrozole and anastrozole are seen in the clinical setting will have to await the results of clinical trials which are currently in progress. PMID- 11384879 TI - MHC class II expression and antigen presentation by human endometrial cells. AB - It has been demonstrated previously that mixed cell suspensions from the female reproductive tract consisting of human epithelial and stromal cells were capable of presenting foreign antigen to autologous T cells. There have been, however, no reported studies examining antigen presentation by isolated epithelial cells from the human female reproductive tract. It is now shown that freshly isolated epithelial cells from the uterine endometrium constitutively express MHC class II antigen and that class II was upregulated on cultured epithelium by interferon gamma (IFNgamma). Using a highly purified preparation, it was demonstrated that these epithelial cells were able to process and present tetanus toxoid recall antigen driving autologous T cell proliferation. Cells isolated from the basolateral sub-epithelium stroma were also potent antigen presenting cells in this model system. Thus, isolated endometrial epithelial cells were able to directly process and present antigen to T cells and may be responsible for the transcytosis and delivery of antigen to professional antigen presenting cells found in the sub-epithelial stroma. PMID- 11384880 TI - Multiple signal transduction pathways mediate interleukin-4-induced 3beta hydroxysteroid dehydrogenase/Delta5-Delta4 isomerase in normal and tumoral target tissues. AB - The 3beta-hydroxysteroid dehydrogenase/Delta5-Delta4 isomerase (3beta-HSD) isoenzymes catalyze an essential step in the formation of all classes of active steroid hormones. We have recently shown that 3beta-HSD type 1 gene expression is specifically induced by interleukin (IL)-4 and IL-13 in several human cancer cell lines and in normal human mammary and prostatic epithelial cells in primary culture. There is evidence that IL-4 stimulates bifurcating signaling pathways in which the Stat6-signal pathway is involved in differentiation and gene regulation, whereas insulin receptor substrate (IRS) proteins mediate the mitogenic action of IL-4. As a matter of fact, we have shown that IL-4-activated Stat6 in all cell lines studied, where IL-4 induced 3beta-HSD type 1 expression but not in those cell lines that failed to respond to IL-4. The mechanism of the induction of 3beta-HSD type 1 gene expression was further characterized in ZR-75 1 human breast cancer cells. We have also found that IL-4 rapidly induced IRS-1 and IRS-2 phosphorylation in these cell lines. Moreover, insulin-like growth factor (IGF)-1 and insulin, which are well known to cause IRS-1 and IRS-2 phosphorylation, increased the stimulatory effect of IL-4 on 3beta-HSD activity. IRS-1 and IRS-2 are adapter molecules that provide docking sites for different SH2 domain-containing proteins, leading to the activation of multiple pathways, such as the phosphatidylinositol (PI) 3-kinase and the mitogen-activated protein (MAP) pathways. The inhibition of IL-4-induced 3beta-HSD expression by PI 3 kinase inhibitors (wortmannin and LY294002) as well as an inhibitor of MAP kinase activation (PD98059), indicates the involvement of those pathways in this response to IL-4. Wortmannin also blocked MAP kinase activation by IL-4, insulin and IGF-1 suggesting that the MAP kinase cascade acts as a downstream effector of PI 3-kinases. Furthermore, we showed that the PKC activator phorbol-12-myristate 13-acetate (PMA) also potentiated the IL-4-induced 3beta-HSD activity, thus suggesting that one signaling molecule that is involved in the signal transduction of the IL-4 action on 3beta-HSD type 1 expression is also a substrate for PKC. Taken together, these findings suggest the existence of a novel mechanism of gene regulation by IL-4. This mechanism would involve in the phosphorylation of IRS-1 and IRS-2, which transduce the IL-4 signal through a PI 3-kinase- and MAP kinase-dependent signaling pathway. However, the inability of IGF-1, insulin and PMA to stimulate 3beta-HSD type 1 expression by themselves in the absence of IL-4 indicates that the multiple pathways downstream of IRS-1 and IRS-2 must act in cooperation with an IL-4-specific signaling molecule, such as the transcription factor Stat6. It is also of interest to note that there also appear to be differences between the regulation of the 3beta-HSD type 1 and type 2 promoters. PMID- 11384882 TI - Tibolone: a steroid with a tissue-specific mode of action. AB - In postmenopausal women tibolone has proved to prevent bone-loss and relieve climacteric symptoms as effectively as estrogens, but it does not stimulate the endometrium and the breast. This clinical profile strongly suggests that tibolone is a compound with tissue-specific action. Tibolone is quickly metabolized into its main active metabolites, 3alpha and 3beta-OH, which are also present in an inactive, sulphated, form. In addition a Delta4-metabolite is found in circulation. The 3-OH-metabolites bind only to the estrogen receptor while the Delta4-isomer shows affinity only to the progesterone and androgen receptors. Tibolone prevents bone loss in a similar way to estrogens. Studies on bone mass using anti-estrogen, antiprogestin and anti-androgen in combination with tibolone, confirmed the sole involvement of the estradiol receptor. Increases in skin temperature as well as vaginal atrophy can be prevented by tibolone in a similar way to estrogens. Breast safety studies showed that tibolone clearly inhibited the growth of tumors in a DMBA model. In breast cell lines, tibolone profoundly inhibited sulphatase activity and an increase in apoptosis and decrease in cell proliferation was found. The stimulation of the endometrium is prevented by the local formation of the Delta4-isomer from tibolone or the 3beta OH-metabolite. We conclude that tibolone acts as a tissue-specific compound by mediating its effects via steroid receptors and enzymatic pathways. This dual effect of tibolone explains it's positive clinical effects on bone, vagina and brain, and avoids stimulation of the endometrium and breast tissue. PMID- 11384881 TI - Alzheimer's disease and estrogen. AB - The preventive effect of estrogen on Alzheimer's disease (AD) has become clear with epidemiological data. Therapeutic effects of estrogen have not yet been established. In this presentation, we report our new basic and clinical data. The estrogen receptor, (ER)alpha, and ERbeta mRNA were investigated in rat brain. Estradiol-17beta (E(2)) treatment following OVX reduced the levels of ERalpha mRNA in the hypothalamus. In the substantia innominata (SI), the number of choline acetyltransferase immunoreacive cells increased significantly in the estrogen treatment rat. The neurons in SI projecting to the forebrain cortex contained ERalpha. Increasing amounts of intracellular calcium, peroxidation, and apoptosis with amyloid beta were suppressed in neuronal cells from rat pheochromocytoma (PC12) cells with E(2). ERalpha cDNA transfected PC 12 cells elaborated more neurite-like processes with E(2). In clinics, we are currently preparing vaginal progesterone tablets, which essentially may concentrate in the endometrium to prevent endometrial cancer, with few general circulation of progesterone inviting less depression. The therapeutic effects of cyclic estrogen, such as its preventive effect, are suggested in these studies, at least on mild AD. PMID- 11384883 TI - Why vegetable recipes are not very spicy. AB - Spices are aromatic plant materials that are used in cooking. Recently it was hypothesized that spice use yields a health benefit: cleansing food of parasites and pathogens before it is eaten, thereby reducing food poisoning and foodborne illnesses. In support, most spices have antimicrobial properties and use of spices in meat-based recipes is greatest in hot climates, where the diversity and growth rates of microorganisms are highest. A critical prediction of the antimicrobial hypothesis is that spices should be used less in preparing vegetables than meat dishes. This is because cells of dead plants are better protected physically and chemically against bacteria and fungi than cells of dead animals (whose immune system ceased functioning at death), so fewer spices would be necessary to make vegetables safe for consumption. We tested this corollary by compiling information on 2129 vegetable-only recipes from 107 traditional cookbooks of 36 countries. Analyses revealed that spice use increased with increasing ambient temperature, but less dramatically than in meat-based recipes. In all 36 countries, vegetable dishes called for fewer spices per recipe than meat dishes; 27 of these differences were significant. Of 41 individual spices, 38 were used less frequently in vegetable recipes; 30 of these differences were significant. Proportions of recipes that called for >1 spice and >1 extremely potent antimicrobial spice also were significantly lower for vegetable dishes. By every measure, vegetable-based recipes were significantly less spicy than meat based recipes. Within-country analyses control for possible differences in spice plant availability and degrees of cultural independence. Results thus strongly support the antimicrobial hypothesis. PMID- 11384884 TI - The evolution of prestige: freely conferred deference as a mechanism for enhancing the benefits of cultural transmission. AB - This paper advances an "information goods" theory that explains prestige processes as an emergent product of psychological adaptations that evolved to improve the quality of information acquired via cultural transmission. Natural selection favored social learners who could evaluate potential models and copy the most successful among them. In order to improve the fidelity and comprehensiveness of such ranked-biased copying, social learners further evolved dispositions to sycophantically ingratiate themselves with their chosen models, so as to gain close proximity to, and prolonged interaction with, these models. Once common, these dispositions created, at the group level, distributions of deference that new entrants may adaptively exploit to decide who to begin copying. This generated a preference for models who seem generally "popular." Building on social exchange theories, we argue that a wider range of phenomena associated with prestige processes can more plausibly be explained by this simple theory than by others, and we test its predictions with data from throughout the social sciences. In addition, we distinguish carefully between dominance (force or force threat) and prestige (freely conferred deference). PMID- 11384885 TI - Maternal trade-off in treating high-risk children. AB - Low birthweight and the infant's health status are expected to strongly influence the child's reproductive value and, thus, the maternal decisions on the amount and timing of investment. A total of 590 Hungarian primiparous mothers giving birth in the late 1980s were recruited for the longitudinal study. Mothers of high-risk infants shortened the duration of breast-feeding and interbirth intervals, compared to those with an infant of higher survival prospects. The most powerful predictor of the length of the lactation period was the infant's weight at birth, whereas birth spacing was significantly influenced by the health status of the older child. Socioeconomic status had a positive effect on maternal care as well, but it did not change the basic pattern of diminishing maternal care as a function of the infants' low reproductive value. The combination of the above factors resulted in a cumulative effect on maternal investment of mothers with handicapped children of various degrees of risk. An attempt has been made to exclude alternative explanations and to discuss the proximate mechanisms of discriminative parental solicitude. PMID- 11384886 TI - New concepts for prevention and treatment of late-life depression. PMID- 11384887 TI - Successful aging. AB - OBJECTIVE: Until now, prospective studies of aging have begun with 50-60-year olds, not adolescents. Premature death, childhood variables, and alcohol abuse have been often ignored, as has successful aging. METHOD: The authors reviewed the existing literature on health in late life in order to highlight that, increasingly, successful aging is not an oxymoron. The present study followed two cohorts of adolescent boys (237 college students and 332 core-city youth) for 60 years or until death. Complete physical examinations were obtained every 5 years and psychosocial data every 2 years. Predictor variables assessed before age 50 included six variables reflecting uncontrollable factors: parental social class, family cohesion, major depression, ancestral longevity, childhood temperament, and physical health at age 50 and seven variables reflecting (at least some) personal control: alcohol abuse, smoking, marital stability, exercise, body mass index, coping mechanisms, and education. The six outcome variables chosen to assess successful aging at age 70-80 included four objectively assessed variables (physical health, death and disability before age 80, social supports, and mental health) and two self-rated variables (instrumental activities of daily living and life enjoyment). RESULTS: Multivariate analysis suggested that "good" and "bad" aging from age 70-80 could be predicted by variables assessed before age 50. More hopeful still, if the seven variables under some personal control were controlled, depression was the only uncontrollable predictor variable that affected the quality of subjective and objective aging. CONCLUSIONS: One may have greater personal control over one's biopsychosocial health after retirement than previously recognized. PMID- 11384888 TI - How do risk factors work together? Mediators, moderators, and independent, overlapping, and proxy risk factors. AB - OBJECTIVE: The authors developed a methodological basis for investigating how risk factors work together. Better methods are needed for understanding the etiology of disorders, such as psychiatric syndromes, that presumably are the result of complex causal chains. METHOD: Approaches from psychology, epidemiology, clinical trials, and basic sciences were synthesized. RESULTS: The authors define conceptually and operationally five different clinically important ways in which two risk factors may work together to influence an outcome: as proxy, overlapping, and independent risk factors and as mediators and moderators. CONCLUSIONS: Classifying putative risk factors into these qualitatively different types can help identify high-risk individuals in need of preventive interventions and can help inform the content of such interventions. These methods may also help bridge the gaps between theory, the basic and clinical sciences, and clinical and policy applications and thus aid the search for early diagnoses and for highly effective preventive and treatment interventions. PMID- 11384889 TI - Depression in the planet's largest ethnic group: the Chinese. AB - OBJECTIVE: The authors reviewed the evidence for the claim that the Chinese tend to deny depression or express it somatically, examined the possible determinants of those characteristics, and explored implications of the findings for the diagnosis and management of depression in China and for psychiatry in the WEST: METHOD: This paper reviews and interprets original studies and literature reviews considering emotional distress, depression, neurasthenia, and somatization in Chinese subjects. RESULTS: Interpretation of the literature is complicated by the considerable heterogeneity among people described as "the Chinese" and by numerous factors affecting collection of data, including issues of illness definition, sampling, and case finding; differences in help-seeking behavior; idiomatic expression of emotional distress; and the stigma of mental illness. Despite difficulties in interpreting the literature, the available data suggest that the Chinese do tend to deny depression or express it somatically. CONCLUSIONS: The existing evidence supports the hypothesis that the Chinese tend to deny depression or express it somatically. However, Western influences on Chinese society and on the detection and identification of depression are likely to have modified the expression of depressive illness quite sharply since the early 1980s. Analyzing these changes may provide useful insight into the evolution of the diagnosis of depression in Western and other cultures. PMID- 11384890 TI - The human genome: susceptibility loci. PMID- 11384891 TI - Treatment of a 50-year-old African American woman whose chronic posttraumatic stress disorder went undiagnosed for over 20 years. PMID- 11384892 TI - Moral treatment at the end of the Oregon Trail: Dr. James C. Hawthorne, 1819 1881. PMID- 11384893 TI - Syndromic validity of apathy in Alzheimer's disease. AB - OBJECTIVE: The study examined the usefulness and clinical correlates of specific diagnostic criteria for apathy in Alzheimer's disease. Whereas apathy is a frequent behavioral change in patients with Alzheimer's disease, the lack of standardized diagnostic criteria may explain the wide discrepancies in estimates of the frequency and demographic and clinical correlates of apathy. METHOD: A consecutive series of 319 patients who met the criteria for probable Alzheimer's disease established by the National Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer's Disease and Related Disorders Association, 117 patients who met the DSM-IV criteria for depression without dementia, and 36 healthy individuals were assessed with a structured psychiatric interview. On the basis of modified Marin's criteria for apathy, they were classified into groups with or without apathy. RESULTS: Apathy was diagnosed in 37% of the 319 Alzheimer's disease patients, compared to none of the healthy comparison subjects. In 24% of the Alzheimer's disease sample, apathy coexisted with either dysthymic disorder or major depressive disorder, whereas 13% had apathy without depression. Apathy was diagnosed in 32% of the depressed nondemented patients, mostly in those with major depressive disorder. Apathy in Alzheimer's disease was significantly associated with severe impairments in activities of daily living and cognitive functions, older age, and poor awareness of behavioral and cognitive changes. CONCLUSIONS: This study provides partial validation of specific clinical criteria for apathy in Alzheimer's disease. PMID- 11384894 TI - Relationship of deep white matter hyperintensities and apolipoprotein E genotype to depressive symptoms in older adults without clinical depression. AB - OBJECTIVE: This study examined whether evidence of cerebrovascular disease in the form of magnetic resonance imaging (MRI) signal hyperintensities in white matter was associated with depressive symptoms in a high-functioning group of normal elderly volunteers. METHOD: Ninety-two community-dwelling elderly individuals participating in a study of white matter hyperintensities (WMHs) in normal aging whose apolipoprotein E (APOE) genotype had been determined completed the Geriatric Depression Scale and received an MRI scan. Univariate analyses of variance were used to examine the relationship between depressive symptoms and the location of WMHs (in deep white matter versus in periventricular white matter) and to determine whether WMHs were more likely to be associated with symptoms of impaired motivation and concentration or with mood symptoms. The effect on depressive symptoms of the interaction between severity of cerebrovascular disease as evidenced by WMHs and APOE genotype was also examined. RESULTS: Hyperintensities in the deep white matter, but not in the periventricular white matter, were associated with depressive symptoms, especially symptoms of impaired motivation, concentration, and decision making. The relationship between deep WMHs and depressive symptoms was especially strong in individuals carrying the APOE-4 allele. CONCLUSIONS: The pattern of depressive symptoms associated with WMHs in this study was similar to the pattern described in the literature as characterizing "vascular" depression in older persons with major depression. The results suggest that cerebrovascular disease may also underlie the depressive symptoms often found in older individuals who are not clinically depressed. PMID- 11384895 TI - The interplay and etiological continuity of neuroticism, difficulties, and life events in the etiology of major and subsyndromal, first and recurrent depressive episodes in later life. AB - OBJECTIVE: Stressful life events, long-term difficulties, and high neuroticism are established risk factors for depression. Less is known about their role in late-life depression, how they modify or mediate one another's effects, and whether this differs between major and subsyndromal, first and recurrent episodes. METHOD: The authors used a prospective case-control design nested in a community survey of elderly subjects that included a measure of neuroticism. They compared 83 survey participants who subsequently developed a depressive episode with 83 randomly selected comparison participants. The authors determined dates of onset, history, and severity of episodes and dates of occurrence and severity of stressful life events and difficulties. RESULTS: Stressful life events did not mediate the effects of high neuroticism and difficulties at onset, possibly because of the uncontrollable nature of common stressful life events in later life. Without both high neuroticism and difficulties, stressful life events did not increase risk. High neuroticism and difficulties increased risk, even without a stressful life event. In the presence of high neuroticism and/or difficulties, the depressogenic effect of stressful life events was substantial, suggesting effect modification. The authors found no evidence to suggest etiological discontinuity between major and subsyndromal episodes. First and recurrent episodes showed a discontinuous pattern of associations. Severe stressful life events had weaker associations, but high neuroticism and mild stressful life events had stronger associations with recurrent than with first episodes. CONCLUSIONS: This study demonstrated the usefulness of a dynamic stress vulnerability model for understanding late-life depression. Evidence was found suggesting etiological discontinuity between first and recurrent but not between major and subsyndromal episodes. PMID- 11384896 TI - Hazardous benzodiazepine regimens in the elderly: effects of half-life, dosage, and duration on risk of hip fracture. AB - OBJECTIVE: While benzodiazepine treatment is known to increase the risk of hip fracture in older populations, controversy persists over which characteristics of benzodiazepine use (e.g., elimination half-life, dosage, duration of use) are most associated with such risks. METHOD: The authors reviewed the health care utilization data of 1,222 hip fracture patients and 4,888 comparison patients frequency matched on the basis of age and gender (all were at least 65 years old). Patients were enrolled in Medicare as well as in the New Jersey Medicaid or Pharmaceutical Assistance to the Aged and Disabled programs. Benzodiazepine use, as well as other covariates, were assessed before the index date (which was either the date of hospital admission for hip fracture surgical repair or, for the comparison subjects, a randomly assigned, frequency-matched date). RESULTS: All benzodiazepine doses > or =3 mg/day in diazepam equivalents significantly increased the adjusted risk of hip fracture by 50%. Significantly increased adjusted risks of hip fracture were seen during the initial 2 weeks of use (60% increase) and after more than 1 month of continuous use (80% increase) but not for 2-4 weeks of continuous use. Use of benzodiazepines other than long-acting agents significantly increased the risk of hip fracture by 50%. CONCLUSIONS: Even at modest doses, including some low doses currently advocated in prescribing guidelines for older patients, treatment with benzodiazepines appears to increase the risk of hip fracture. Patients appear to be particularly vulnerable immediately after initiating therapy and after more than 1 month of continuous use. Benzodiazepines with shorter half-lives appear to be no safer than longer half-life agents. Clinicians should be aware of these risks and weigh them against potential benefits when prescribing for elderly patients. PMID- 11384897 TI - Changes in regional brain glucose metabolism measured with positron emission tomography after paroxetine treatment of major depression. AB - OBJECTIVE: Depression is commonly associated with frontal hypometabolic activity accompanied by hypermetabolism in certain limbic regions. It is unclear whether successful antidepressant treatments reverse these abnormalities or create new resting levels of metabolism. The aim of the present study was to assess the effects of successful paroxetine treatment on regional glucose metabolism in patients with major depression. METHOD: Positron emission tomography with [(18)F]fluorodeoxyglucose was performed on 13 male patients before and after 6 weeks of paroxetine therapy. Resting state scans were also acquired under similar conditions in 24 healthy male subjects for comparison. RESULTS: After successful paroxetine therapy, increased glucose metabolism occurred in dorsolateral, ventrolateral, and medial aspects of the prefrontal cortex (left greater than right), parietal cortex, and dorsal anterior cingulate. Areas of decreased metabolism were noted in both anterior and posterior insular regions (left) as well as right hippocampal and parahippocampal regions. In comparison to metabolism levels in a group of healthy volunteers, the increase in prefrontal metabolic activity represented a normalization of previously reduced metabolic activity, whereas the reduction in pregenual anterior cingulate activity represented a decrease from previously elevated metabolic levels. CONCLUSIONS: These results provide further support for a dysfunction in cortical-limbic circuitry in depression, which is at least partly reversed after successful paroxetine treatment. PMID- 11384898 TI - Double-blind, placebo-controlled comparison of imipramine and paroxetine in the treatment of bipolar depression. AB - OBJECTIVE: This study compared the efficacy and safety of paroxetine and imipramine with that of placebo in the treatment of bipolar depression in adult outpatients stabilized on a regimen of lithium. METHOD: In a double-blind, placebo-controlled study, 117 outpatients with DSM-III-R bipolar disorder, depressive phase, were randomly assigned to treatment with paroxetine (N=35), imipramine (N=39), or placebo (N=43) for 10 weeks. In addition to lithium monotherapy, patients may have received either carbamazepine or valproate in combination with lithium for control of manic symptoms. Patients were stratified on the basis of trough serum lithium levels determined at the screening visit (high: >0.8 meq/liter; low: 0.05). Therefore, in age-adjusted analyses, smaller LDL particle size was associated with MI in young women, but the risk was attenuated after adjustments for metabolic factors related to both LDL particle size and MI. PMID- 11384950 TI - Changes in body mass index prior to baseline among participants who are ill or who die during the early years of follow-up. AB - The association between body mass index (weight (kg)/height (m)(2)) and mortality may be confounded by preexisting illness. A method commonly used to control for this confounding is the exclusion of participants who have certain diseases at baseline and/or those who die during the early years of follow-up. The authors used data from the Atherosclerosis Risk in Communities (ARIC) Study (n = 14,088) to determine whether participants identified by these criteria had different changes in body mass index than other participants. Weight change was measured over a 3-year interval between study entry (1987-1989) and reexamination (1990 1993), and information on vital status was collected over the subsequent 5 years. Mean change in body mass index was -0.54 (95% confidence interval (CI): -0.90, 0.12) among participants who died in the first year of follow-up, -0.03 (95% CI: 0.18, 0.12) among those who died in the first 4 years of follow-up, and 0.36 (95% CI: 0.33, 0.39) among those who survived for at least 5 years. Participants who died during the first 4 years were over twice as likely as survivors to have changed from the obese category (body mass index >or=30) to the nonobese category (odds ratio = 2.14; 95% CI: 1.44, 3.17). Mean change in body mass index prior to baseline was not different among ill participants compared with those who were healthy, but the odds of converting from obese to nonobese were higher in ill participants than in healthy ones (odds ratio = 1.29; 95% CI: 1.01, 1.67). PMID- 11384951 TI - Measures of maternal tobacco exposure and infant birth weight at term. AB - This study was undertaken to determine the relation between self-reported number of cigarettes smoked per day and urine cotinine concentration during pregnancy and to examine the relations between these two measures of tobacco exposure and birth weight. Data were obtained from the Smoking Cessation in Pregnancy project, conducted between 1987 and 1991. Cigarette smoking information and urine cotinine concentration were collected for 3,395 self-reported smokers who were receiving prenatal care at public clinics in three US states (Colorado, Maryland, and Missouri) and who delivered term infants. General linear models were used to quantify urine cotinine variability explained by the number of cigarettes smoked per day and to generate mean adjusted birth weights for women with different levels of tobacco exposure. Self-reported number of cigarettes smoked per day explained only 13.9% of the variability in urine cotinine concentration. Birth weight declined as tobacco exposure increased; however, the relation was not linear. The sharpest declines in birth weight occurred at low levels of exposure. Furthermore, urine cotinine concentration did not explain more variability in birth weight than did number of cigarettes smoked. These findings should be considered by researchers studying the effects of smoking reduction on birth outcomes. PMID- 11384952 TI - Neural tube defects in relation to use of folic acid antagonists during pregnancy. AB - Periconceptional folic acid supplementation reduces the risk of neural tube defects (NTDs). To determine whether periconceptional exposure to folic acid antagonists (FAAs) might therefore increase the risk of NTDs, the authors examined data from an ongoing case-control study of birth defects (1979-1998) in the United States and Canada. They compared data on 1,242 infants with NTDs (spina bifida, anencephaly, and encephalocele) with data from a control group of 6,660 infants with malformations not related to vitamin supplementation. Mothers were interviewed within 6 months of delivery about demographic, reproductive, medical, and behavioral factors and about medication use. The adjusted odds ratios of NTDs related to exposure to FAAs (including carbamazepine, phenobarbital, phenytoin, primidone, sulfasalazine, triamterene, and trimethoprim) during the first or second months after the last menstrual period, compared with no use in either month, were 2.8 (95% confidence interval: 1.7, 4.6) for FAAs as a group, 4.8 (95% confidence interval: 1.5, 16.1) for trimethoprim (based on five exposed cases), and 6.9 (95% confidence interval: 1.9, 25.7) for carbamazepine (six exposed cases). These results are adjusted for region, interview year, periconceptional folic acid supplementation, maternal age, weight, education, and infections early in pregnancy. These findings suggest that a number of FAAs may increase NTD risk, and they provide estimates of risk for selected drugs. PMID- 11384953 TI - Trends in nutrient intake of 10-year-old children over two decades (1973-1994) : the Bogalusa Heart Study. AB - Dietary intakes of 10-year-old children were examined in seven cross-sectional surveys to observe secular trends in nutrient intake and food consumption patterns over 2 decades (1973-1994). Total energy intake remained unchanged from 1973 to 1994. However, when expressed as energy per kilogram body weight, intake decreased from 65.5 kcal in 1973 to 55.4 kcal in 1994 because children's weight increased. A significant trend was noted in ponderal index, which increased from 12.31 (1973-1974) to 13.71 (1992-1994), with an actual weight gain of 1.45 kg from 1973 to 1979 and 2.71 kg from 1981 to 1994. Linear trends also were noted for total fat (negative), saturated fat (negative), dietary cholesterol (negative), polyunsaturated fat (positive), and total carbohydrate (positive). There was a significant increase in percent energy from protein and carbohydrate and a significant decrease in percent energy from fat, primarily saturated and monounsaturated fat. Trends in nutrient intakes of children reflected trends in food consumption. The percentage of total fat from fats/oils, mixed meats, eggs, milk, pork, and desserts decreased, while that from poultry, cheese, and snacks increased. Although more children met dietary recommendations for total fat, saturated fat, and dietary cholesterol, the vast majority continued to exceed prudent diet recommendations. PMID- 11384954 TI - Cancer incidence among workers in the Norwegian silicon carbide industry. AB - The presence of silicon carbide (SiC) fibers in the SiC smelter work environment has suggested a possible cancer hazard. The authors studied cancer incidence among 2,620 men employed for more than 6 months in three Norwegian SiC smelters. Follow-up from 1953 to 1996 revealed an overall excess risk of lung cancer (standardized incidence ratio = 1.9, 95% confidence interval (CI): 1.5, 2.3) and an elevated risk of stomach cancer (standardized incidence ratio = 1.5, 95% CI: 1.1, 2.0). Both standardized incidence ratio and Poisson regression analyses showed that lung cancer risk increased according to cumulative exposure to total dust, SiC fibers, SiC particles, and crystalline silica. The standardized incidence ratio for the upper SiC fiber exposure category was 3.5 (95% CI: 2.1, 5.6) when exposure was lagged by 20 years, while the Poisson regression analysis showed a rate ratio of 4.4 (95% CI: 2.1, 9.0). Smoking did not seem to be an important confounder. The excess risk of lung cancer may be explained by exposure to SiC fibers, but a strong correlation between the different exposures made it difficult to distinguish between them. PMID- 11384955 TI - Sources of variance in daily physical activity levels in the seasonal variation of blood cholesterol study. AB - The authors examined sources of variance in self-reported physical activity in a cohort of healthy adults (n = 580) from Worcester, Massachusetts (the Seasonal Variation of Blood Cholesterol Study, 1994-1998). Fifteen 24-hour physical activity recalls of total, occupational, and nonoccupational activity (metabolic equivalent-hours/day) were obtained over 12 months. Random effects models were employed to estimate variance components for subject, season, day of the week, and residual error, from which the number of days of assessment required to achieve 80% reliability was estimated. The largest proportional source of variance in total and nonoccupational activity was within-subject variance (50 60% of the total). Differences between subjects accounted for 20-30% of the overall variance in total activity, and seasonal and day-of-the-week effects accounted for 6% and 15%, respectively. For total activity, 7-10 days of assessment in men and 14-21 days of assessment in women were required to achieve 80% reliability. For nonoccupational activity, 21-28 days of assessment were required. This study is among the first to have examined the sources of variance in daily physical activity levels in a large population of adults using 24-hour physical activity recall. These findings provide insight for understanding the strengths and limitations of short term and long term physical activity assessments employed in epidemiologic studies. PMID- 11384956 TI - Statistical issues in analyzing 24-hour dietary recall and 24-hour urine collection data for sodium and potassium intakes. AB - Dietary recalls and urine assays provide different metrics for assessing sodium and potassium intakes. Means, variances, and correlations of data obtained from these two modes of measurement differ. Pooling of these data is not straightforward, and results from studies employing the different modes may not be comparable. To explore differences between these metrics, the authors used data from the Trial of Nonpharmacologic Intervention in the Elderly (TONE), which included repeated standardized 24-hour dietary recalls and 24-hour urine collections administered over 3 years of follow-up, to estimate sodium and potassium intakes. The authors examined data from 341 control participants assigned to usual care that were collected between August 1992 and December 1995. Dietary recalls yielded estimates of sodium intake that averaged 22% less than those from urine assays and estimates of potassium intake that averaged 16% greater than those from urine assays. Sodium intake estimates were less repeatable (r = 0.22 for diet; r = 0.30 for urine) than potassium intake estimates (r = 0.49 for diet; r = 0.50 for urine), making relations with outcomes more difficult to characterize. Overall, the performance of the two measurement modes was fairly similar across demographic subgroups. Errors in separate estimations of long term sodium and potassium intakes using short term data were strongly correlated, more strongly than the underlying long term intakes of these electrolytes. Because of the correlated measurement error, estimated regression coefficients for linear models including both electrolytes as predictors may be confounded such that the separate relations between these nutrients and outcomes such as blood pressure cannot be reliably estimated by common analytical strategies. PMID- 11384957 TI - Evaluation of two putative susceptibility loci for oral clefts in the Danish population. AB - Previous studies suggest that the risk of nonsyndromic cleft lip with or without cleft palate (CL+/-P) and isolated cleft palate (CP) is influenced by genetic variation at several loci and that the relation between specific genetic variants and disease risk may be modified by environmental factors. The present study evaluated potential associations between CL+/-P and CP and two putative clefting susceptibility loci, MSX1 and TGFB3, using data from a nationwide case-control study conducted in Denmark from 1991 to 1994. The potential effects of interactions between these genes and two common environmental exposures, first trimester exposure to maternal cigarette smoke and alcohol intake, were also examined. Analyses of these data provide evidence of an association between the risk of CP and variation at the TGFB3 locus. However, there is no evidence that the risk of CL+/-P or CP is influenced by gene-environment interactions involving MSX1 or TGFB3 and either first trimester exposure to maternal cigarette smoke or alcohol consumption. PMID- 11384958 TI - Commentary: facing the challenge of gene-environment interaction: the two-by-four table and beyond. AB - As a result of the Human Genome Project, epidemiologists can study thousands of genes and their interaction with the environment. The challenge is how to best present and analyze such studies of multiple genetic and environmental factors. The authors suggest emphasizing the fundamental core of gene-environment interaction-the separate assessment of the effects of individual and joint risk factors. In the simple analysis of one genotype and an exposure (both dichotomous), such study can be summarized in a two-by-four table. The advantages of such a table for data presentation and analysis are many: The table displays the data efficiently and highlights sample size issues; it allows for evaluation of the independent and joint roles of genotype and exposure on disease risk; and it emphasizes effect estimation over model testing. Researchers can easily estimate relative risks and attributable fractions and test different models of interaction. The two-by-four table is a useful tool for presenting, analyzing, and synthesizing data on gene-environment interaction. To highlight the role of gene-environment interaction in disease causation, the authors propose that the two-by-four table is the fundamental unit of epidemiologic analysis. PMID- 11384959 TI - Recall bias in self-reported melanoma risk factors. AB - The evidence implicating sun exposure in the etiology of melanoma derives largely from case-control studies in which the retrospective assessment of sun exposure suggests potential for significant recall bias. Previous attempts at characterizing and quantifying that bias have had significant methodological limitations. In the International Twin Study, a case-control study of melanoma risk factors in twins conducted from 1980 to 1991, the authors asked melanoma cases and their co-twins to quantify their own exposures and asked which twin had the greater exposure. Recall bias was investigated by assuming that, if bias had occurred, the odds ratio based on the case's response would differ significantly from the odds ratio based on the co-twin's response. Case-derived odds ratios were higher than the odds ratios for the controls for sunbathing in childhood and adulthood and for mole frequency and freckling in childhood, suggesting some recall bias. The odds ratios for ease of burning and tanning appeared unbiased. The belief that sunlight was a cause of melanoma appeared related to an increased odds ratio for sunbathing as a child. There is a continuing need to carefully assess recall bias in the study of melanoma risk factors. PMID- 11384960 TI - Re: "Estimating particulate matter-mortality dose-response curves and threshold levels: an analysis of daily time-series for the 20 largest US cities". PMID- 11384961 TI - Re: "Occupational exposure and lung cancer risk: a population-based case-referent study in Sweden". PMID- 11384963 TI - BRCA1 is a selective co-activator of 14-3-3 sigma gene transcription in mouse embryonic stem cells. AB - BRCA1 gene is a tumor suppressor for breast and ovarian cancers with the putative role in DNA repair and transcription. To characterize the role of BRCA1 in transcriptional regulation, we analyzed gene expression profiles of mouse embryonic stem cells deficient in BRCA1 using microarray technology. We found that loss of BRCA1 correlated with decreased expression of several groups of genes including stress response genes, cytoskeleton genes, and genes involved in protein synthesis and degradation. Previous study showed that BRCA1 is a transcriptional co-activator of p53 protein; however the majority of p53 target genes remained at the same expression levels in BRCA1 knockout cells as in the wild type cells. The only p53 target gene down-regulated with the loss of BRCA1 was 14-3-3 sigma, a major G(2)/M checkpoint control gene. Similar to cells with decreased 14-3-3 sigma activity, BRCA1-deficient cells were unable to sustain G(2)/M growth arrest after exposure to ionizing radiation. We find that BRCA1 induction of 14-3-3 sigma requires the presence of wild type p53 and can be regulated by a minimal p53 response element. PMID- 11384964 TI - Kinetic and thermodynamic analysis of the interactions of 23-residue peptides with endotoxin. AB - Many naturally occurring peptides exhibit lipopolysaccharide binding properties. In this work we describe the endotoxin binding properties of a series of 23 residue peptides based on the sequence corresponding to the antisense strand of the magainin gene. Biochemical and biophysical characterization of these peptides reveals that they have the tendency to perturb both the inner and outer membranes of test pathogens. Structurally these peptides are amphiphilic and adopt helical conformations in membranes. Three of the seven peptides tested have high affinities for endotoxin that approach the values shown by polymyxin B, a cyclic cationic acylated decapeptide, which is used clinically in treating extreme cases of sepsis. The kinetic parameters obtained using stopped-flow methods and BIAcore analysis, when considered in conjunction with the isothermal titration calorimetry-derived thermodynamic parameters, allow us to highlight the key structural features essential for lipopolysaccharide (LPS) recognition by these peptides. The studies stress the role of ionic forces in the initial recognition of LPS. The fortification of the strength of these ionic charges increases affinity for LPS, whereas the hydrophobic residues involved in interactions are more amenable to disruptions in contiguity. Peptides that improve these features further are expected to perform better as endotoxin-neutralizing agents. PMID- 11384965 TI - Fas-associated death domain protein (FADD) and caspase-8 mediate up-regulation of c-Fos by Fas ligand and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) via a FLICE inhibitory protein (FLIP)-regulated pathway. AB - Fas, a death domain-containing member of the tumor necrosis factor receptor family and its ligand FasL have been predominantly studied with respect to their capability to induce cell death. However, a few studies indicate a proliferation inducing signaling activity of these molecules too. We describe here a novel signaling pathway of FasL and the tumor necrosis factor-related apoptosis inducing ligand (TRAIL) that triggers transcriptional activation of the proto oncogene c-fos, a typical target gene of mitogenic pathways. FasL- and TRAIL mediated up-regulation of c-Fos was completely dependent on the presence of Fas associated death domain protein (FADD) and caspase-8, but caspase activity seemed to be dispensable as a pan inhibitor of caspases had no inhibitory effect. Upon overexpression of the long splice form of cellular FADD-like interleukin-1 converting enzyme (FLICE) inhibitory protein (cFLIP) in Jurkat cells, FasL- and TRAIL-induced up-regulation of c-Fos was almost completely blocked. The short splice form of FLIP, however, showed a rather stimulatory effect on c-Fos induction. Together these data demonstrate the existence of a death receptor induced, FADD- and caspase-8-dependent pathway leading to c-Fos induction that is inhibited by the long splice form FLIP-L. PMID- 11384966 TI - Reconstitution of acid secretion in digitonin-permeabilized rabbit gastric glands. Identification of cytosolic regulatory factors. AB - When isolated rabbit gastric glands were permeabilized with digitonin, they lost their ability to secrete acid, as monitored by [14C]aminopyrine accumulation, and they never recovered by supplement with cytosol prepared from gastric mucosa. However, the permeabilized glands elicited acid secretion when brain cytosol was supplemented. Fractionation of gastric cytosol by gel filtration revealed that the fraction at 30 kDa stimulated permeabilized glands by itself, whereas the 200 kDa fraction potently inhibited brain cytosol-stimulated acid secretion. Brain cytosol contained only the former stimulatory factor. With further gel filtration, the 30-kDa activator was separated into two components, 20 kDa (peak 1) and 1.8 kDa (peak 2), both of which are necessary for full activity. We purified peak 1 from bovine brain, and phosphatidylinositol transfer protein (PITP) was identified as the main component of the activity. The stimulating activity in brain and gastric mucosa correlated with the contents of PITP, and recombinant PITP mimicked the effect of peak 1, suggesting that PITP is one of the essential components in gastric acid secretion. When gastric glands were stimulated, the inhibitory activity, but not stimulatory activity, in the cytosol was increased. This suggests a regulatory mechanism such as stimulation translocates the inhibitory component from the secretory site on the membrane to cytosol. These results demonstrate a high degree of usefulness for our present model, the reconstituted digitonin-permeabilized gastric glands. PMID- 11384967 TI - The histone acetyltransferase, hGCN5, interacts with and acetylates the HIV transactivator, Tat. AB - Factor acetyltransferase activity associated with several histone acetyltransferases plays a key role in the control of transcription. Here we report that hGCN5, a well known histone acetyltransferase, specifically interacts with and acetylates the human immunodeficiency virus type 1 (HIV-1) transactivator protein, Tat. The interaction between Tat and hGCN5 is direct and involves the acetyltransferase and the bromodomain regions of hGCN5, as well as a limited region of Tat encompassing the cysteine-rich domain of the protein. Tat lysines 50 and 51, target of acetylation by p300/CBP, were also found to be acetylated by hGCN5. The acetylation of these two lysines by p300/CBP has been recently shown to stimulate Tat transcriptional activity and accordingly, we have found that hGCN5 can considerably enhance Tat-dependent transcription of the HIV 1 long terminal repeat. These data highlight the importance of the acetylation of lysines 50 and 51 in the function of Tat, since different histone acetyltransferases involved in distinct signaling pathways, GCN5 and p300/CBP, converge to acetylate Tat on the same site. PMID- 11384968 TI - Signaling states of rhodopsin: absorption of light in active metarhodopsin II generates an all-trans-retinal bound inactive state. AB - Absorption of light in rhodopsin leads through 11-cis- and all-trans-retinal isomerization, proton transfers, and structural changes to the active G-protein binding meta-II state. When meta-II is photolysed by blue light absorption, the activating pathway is apparently reverted, and rhodopsin is photoregenerated. However, the product formed, a P subspecies with A(max) = 500 nm (P(500)), is different from the ground state based on the following observations: (i) the ground state fingerprint of 11-cis-retinal does not appear in the infrared spectra, although the proton transfers and structural changes are reverted; (ii) extraction of the retinal from P(500) does not yield the expected stoichiometric amount of 11-cis-retinal but predominantly yields all-trans-retinal; (iii) the infrared spectrum of P(500) is similar to the classical meta-III intermediate, which arises from meta-II by thermal decay; and (iv) both P(500) and meta-III can be photoconverted to meta-II with the same changes in the infrared spectrum and without a significant change in the isomerization state of the extracted chromophore. The data indicate the presence of a "second switch" between active and inactive conformations that operates by photolysis but without isomerization around the C(11)-C(12) double bond. This emphasizes the exclusivity of the ground state, which is only accessible by the metabolic regeneration with 11-cis retinal. PMID- 11384969 TI - Molecular characterization of calmodulin trapping by calcium/calmodulin-dependent protein kinase II. AB - Autophosphorylation of alpha-Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II) at Thr(286) results in calmodulin (CaM) trapping, a >10,000-fold decrease in the dissociation rate of CaM from the enzyme. Here we present the first site-directed mutagenesis study on the dissociation of the high affinity complex between CaM and full-length CaM kinase II. We measured dissociation kinetics of CaM and CaM kinase II proteins by using a fluorescently modified CaM that is sensitive to binding to target proteins. In low [Ca(2+)], the phosphorylated mutant kinase F293A and the CaM mutant E120A/M124A exhibited deficient trapping compared with wild-type. In high [Ca(2+)], the CaM mutations E120A, M124A, and E120A/M124A and the CaM kinase II mutations F293A, F293E, N294A, N294P, and R297E increased dissociation rate constants by factors ranging from 2.3 to 116. We have also identified residues in CaM and CaM kinase II that interact in the trapped state by mutant cycle-based analysis, which suggests that interactions between Phe(293) in the kinase and Glu(120) and Met(124) in CaM specifically stabilize the trapped CaM-CaM kinase II complex. Our studies further show that Phe(293) and Asn(294) in CaM kinase II play dual roles, because they likely destabilize the low affinity state of CaM complexed to unphosphorylated kinase but stabilize the trapped state of CaM bound to phosphorylated kinase. PMID- 11384970 TI - Channeling during prothrombin activation. AB - The plasma zymogen prothrombin (II) is converted to the clotting enzyme thrombin (IIa) by two prothrombinase-catalyzed proteolytic cleavages. Thus, two intermediates, meizothrombin (mIIa) and prethrombin-2 (P2), are possible on the reaction pathway. Measurements of the time courses of II, mIIa, P2, and IIa suggested a channeling phenomenon, whereby a portion of the II is converted directly to IIa without free mIIa and P2 as obligatory intermediates. Evidence for this was that the maximum rate of IIa formation preceded the maximum in the level of either intermediate. In addition, analysis of the data according to a model that included two parallel pathways through mIIa and P2 indicated that about 40% of the II consumed did not yield free mIIa or P2. Further studies were carried out in which II was continuously infused in a reactor at a constant rate. Under these conditions II, mIIa, and P2 reached constant steady-state levels, and IIa was produced at a constant rate, equal to that of II infusion. During the steady state, traces of II, mIIa, and P2 were introduced as radiolabels. Time courses of isotope consumption were first order, thus allowing the rates of consumption of II, mIIa, and P2 to be calculated. Under these conditions the rate of II consumption equaled the rate of IIa formation. Rates of consumption of the free intermediates, however, were only 22 (mIIa) and 15% (P2), respectively, of the rate of thrombin formation. Thus, both the time course experiments and the steady-state experiments indicate that an appreciable fraction of II is channeled directly to IIa without proceeding through the free intermediates mIIa and P2. PMID- 11384971 TI - Fibroblast growth factor inhibits chondrocytic growth through induction of p21 and subsequent inactivation of cyclin E-Cdk2. AB - Fibroblast growth factor (FGF) and its receptor (FGFR) are thought to be negative regulators of chondrocytic growth, as exemplified by achondroplasia and related chondrodysplasias, which are caused by constitutively active mutations in FGFR3. To understand the growth-inhibitory mechanisms of FGF, we analyzed the effects of FGF2 on cell cycle-regulating molecules in chondrocytes. FGF2 dramatically inhibited proliferation of rat chondrosarcoma (RCS) cells and arrested their cell cycle at the G(1) phase. FGF2 increased p21 expression in RCS cells, which assembled with the cyclin E-Cdk2 complexes, although the expression of neither cyclin E nor Cdk2 increased. In addition, the kinase activity of immunoprecipitated cyclin E or Cdk2, assessed with retinoblastoma protein (pRb) as substrate, was dramatically reduced by FGF-2. Moreover, FGF2 shifted pRb to its underphosphorylated, active form in RCS cells. FGF2 not only induced p21 protein expression in proliferating chondrocytes in mouse fetal limbs cultured in vitro but also decreased their proliferation as assessed by the expression of histone H4 mRNA, a marker for cells in S phase. Furthermore, inhibitory effects of FGF2 on chondrocytic proliferation were partially reduced in p21-null limbs, compared with those in wild-type limbs in vitro. Taken together, FGF's growth inhibitory effects of chondrocytes appear to be mediated at least partially through p21 induction and the subsequent inactivation of cyclin E-Cdk2 and activation of pRb. PMID- 11384973 TI - CRHSP-28 regulates Ca(2+)-stimulated secretion in permeabilized acinar cells. AB - CRHSP-28 is a Ca(2+)-regulated heat-stable phosphoprotein, abundant in the apical cytoplasm of epithelial cells that are specialized in exocrine protein secretion. To define a functional role for the protein in pancreatic secretion, recombinant CRHSP-28 (rCRHSP-28) was introduced into streptolysin-O-permeabilized acinar cells, and amylase secretion in response to elevated Ca(2+) was determined. Secretion was enhanced markedly by rCRHSP-28 over a time course that closely corresponded with the loss of the native protein from the intracellular compartment. No effects of rCRHSP-28 were detected until approximately 50% of the native protein was lost from the cytosol. Secretion was enhanced by rCRHSP-28 over a physiological range of Ca(2+) concentrations with 2-3-fold increases in amylase release occurring in response to low micromolar levels of free Ca(2+). Further, rCRHSP-28 augmented secretion in a concentration-dependent manner with minimal and maximal effects occurring at 1 and 25 microg/ml, respectively. Covalent cross-linking experiments demonstrated that native CRHSP-28 was present in a 60-kDa complex in cytosolic fractions and in a high molecular mass complex in particulate fractions, consistent with the slow leak rate of the protein from streptolysin-O-permeabilized cells. Probing acinar lysates with rCRHSP-28 in a gel-overlay assay identified two CRHSP-28-binding proteins of 35 (pp35) and 70 kDa (pp70). Interestingly, preparation of lysates in the presence of 1 mm Ca(2+) resulted in a marked redistribution of both proteins from a cytosolic to a Triton X-100-insoluble fraction, suggesting a Ca(2+)-sensitive interaction of these proteins with the acinar cell cytoskeleton. In agreement with our previous study immunohistochemically localizing CRHSP-28 around secretory granules in acinar cells, gel-overlay analysis revealed pp70 copurified with acinar cell secretory granule membranes. These findings demonstrate an important cell physiological function for CRHSP-28 in the Ca(2+)-regulated secretory pathway of acinar cells. PMID- 11384972 TI - Structural characterization of heparan sulfate and chondroitin sulfate of syndecan-1 purified from normal murine mammary gland epithelial cells. Common phosphorylation of xylose and differential sulfation of galactose in the protein linkage region tetrasaccharide sequence. AB - Syndecan-1, present on the surfaces of normal murine mammary gland epithelial cells, is a transmembrane hybrid proteoglycan, which bears glycosaminoglycan (GAG) side chains of heparan sulfate (HS) and chondroitin sulfate (CS). Purified syndecan-1 ectodomains were analyzed for disaccharide composition and the GAG protein linkage region after digestion with bacterial lyases. The HS chains contained predominantly a nonsulfated unit with smaller proportions of two monosulfated, two disulfated, and a trisulfated unit, whereas CS chains were demonstrated for the first time to bear GlcUA-GalNAc(4-O-sulfate) as a major component as well as GlcUA-GalNAc, GlcUA-GalNAc(6-O-sulfate), and an E disaccharide unit GlcUA-GalNAc(4,6-O-disulfate) as minor yet appreciable components. Two kinds of linkage region tetrasaccharides, GlcUA-Gal-Gal-Xyl and GlcUA-Gal-Gal-Xyl(2-O-phosphate), were found for the HS chains in a molar ratio of 55:45. In marked contrast, an additional sulfated tetrasaccharide, GlcUA-Gal(4 O-sulfate)-Gal-Xyl, was demonstrated only for the CS chains, and the unmodified phosphorylated and sulfated components were present at a molar ratio of 55:26:19. The present study thus provided conclusive evidence for the hypothesis that 4-O sulfation of Gal is peculiar to CS chains in contrast to the phosphorylation of Xyl, which is common to both HS and CS chains. These modifications may be required for biosynthetic maturation of the linkage region tetrasaccharide sequence, which is a prerequisite for creating the repeating disaccharide region of GAG chains and/or biosynthetic selective chain assembly of CS and HS chains. PMID- 11384974 TI - Overexpression of monomeric and multimeric GIRK4 subunits in rat atrial myocytes removes fast desensitization and reduces inward rectification of muscarinic K(+) current (I(K(ACh))). Evidence for functional homomeric GIRK4 channels. AB - K(+) channels composed of G-protein-coupled inwardly rectifying K(+) channel (GIRK) (Kir3.0) subunits are expressed in cardiac, neuronal, and various endocrine tissues. They are involved in inhibiting excitability and contribute to regulating important physiological functions such as cardiac frequency and secretion of hormones. The functional cardiac (K((ACh))) channel activated by G(i)/G(o)-coupled receptors such as muscarinic M(2) or purinergic A(1) receptors is supposed to be composed of the subunits GIRK1 and GIRK4 in a heterotetrameric (2:2) fashion. In the present study, we have manipulated the subunit composition of the K((ACh)) channels in cultured atrial myocytes from hearts of adult rats by transient transfection of vectors encoding for GIRK1 or GIRK4 subunits or GIRK4 concatemeric constructs and investigated the effects on properties of macroscopic I(K(ACh)). Transfection with a GIRK1 vector did not cause any measurable effect on properties of I(K(ACh)), whereas transfection with a GIRK4 vector resulted in a complete loss in desensitization, a reduction of inward rectification, and a slowing of activation. Transfection of myocytes with a construct encoding for a concatemeric GIRK4(2) subunit had similar effects on desensitization and inward rectification. Following transfection of a tetrameric construct (GIRK4(4)), these changes in properties of I(K(ACh)) were still observed but were less pronounced. Heterologous expression in Chinese hamster ovary cells and human embryonic kidney 293 cells of monomeric, dimeric, and tetrameric GIRK4 resulted in robust currents activated by co-expressed A(1) and M(2) receptors, respectively. These data provide strong evidence that homomeric GIRK4 complexes form functional G(beta)gamma gated ion channels and that kinetic properties of GIRK channels, such as activation rate, desensitization, and inward rectification, depend on subunit composition. PMID- 11384976 TI - Initiation of HIV-1 reverse transcription is regulated by a primer activation signal. AB - Reverse transcription of the human immunodeficiency virus type 1 (HIV-1) RNA genome appears to be strictly regulated at the level of initiation. The primer binding site (PBS), at which the tRNA(3)(Lys) molecule anneals and reverse transcription is initiated, is present in a highly structured region of the untranslated leader RNA. Detailed mutational analysis of the U5 leader stem identified a sequence motif in the U5 region that is critical for activation of the PBS-bound tRNA(3)(Lys) primer. This U5 motif, termed the primer activation signal (PAS), may interact with the TPsiC arm of the tRNA(3)(Lys) primer, similar to the additional interaction proposed for the genome of Rous sarcoma virus and its tRNA(Trp) primer. This suggests that reverse transcription is regulated by a common mechanism in all retroviruses. In HIV-1, the PAS is masked through base pairing in the U5 leader stem. This provides a mechanism for positive and negative regulation of reverse transcription. Based on structure probing of the mutant and wild-type RNAs, an RNA secondary structure model is proposed that juxtaposes the critical PAS and PBS motifs. PMID- 11384977 TI - Significant effect of the N-terminal region of the mitochondrial ADP/ATP carrier on its efficient expression in yeast mitochondria. AB - The low-level expression of the bovine heart mitochondrial ADP/ATP carrier (bovine type 1 ADP/ATP carrier (bAAC1)) in the yeast mitochondrial membrane is significantly improved by replacement of its N-terminal region with corresponding regions of the yeast type 1 and 2 carriers (yAAC1 and yAAC2) (Hashimoto, M., Shinohara, Y., Majima, E., Hatanaka, T., Yamazaki, N., and Terada, H. (1999) Biochim. Biophys. Acta 1409, 113--124). To understand why the bAAC1 chimeras were highly expressed in yeast mitochondria, we examined the effects of the length and sequence of the N-terminal region extending into the cytosol on the expression of bAAC1 and yAAC2 derivatives in yeast mitochondria. For this, their N-terminal regions were replaced with peptide fragments of various lengths and sequences derived from those of bAAC1, yAAC1, and yAAC2. We found that a specific amino acid sequence and a definite length of the N-terminal region of yAAC2 were required for high expression of bAAC1 and yAAC2 in yeast mitochondria. We also examined the steady-state transcript levels and expression of these derivatives in whole yeast cells. Based on our results, we discuss the role of the N-terminal region in efficient expression of bAAC1 and yAAC2 in yeast mitochondria. PMID- 11384975 TI - Myosin ii light chain phosphorylation regulates membrane localization and apoptotic signaling of tumor necrosis factor receptor-1. AB - Activation of myosin II by myosin light chain kinase (MLCK) produces the force for many cellular processes including muscle contraction, mitosis, migration, and other cellular shape changes. The results of this study show that inhibition or potentiation of myosin II activation via over-expression of a dominant negative or wild type MLCK can delay or accelerate tumor necrosis factor-alpha (TNF) induced apoptotic cell death in cells. Changes in the activation of caspase-8 that parallel changes in regulatory light chain phosphorylation levels reveal that myosin II motor activities regulate TNF receptor-1 (TNFR-1) signaling at an early step in the TNF death signaling pathway. Treatment of cells with either ionomycin or endotoxin (lipopolysaccharide) leads to activation of myosin II and increased translocation of TNFR-1 to the plasma membrane independent of TNF signaling. The results of these studies establish a new role for myosin II motor activity in regulating TNFR-1-mediated apoptosis through the translocation of TNFR-1 to or within the plasma membrane. PMID- 11384978 TI - The putative tumor suppressor LRP1B, a novel member of the low density lipoprotein (LDL) receptor family, exhibits both overlapping and distinct properties with the LDL receptor-related protein. AB - The low density lipoprotein receptor-related protein-deleted in tumor (LRP1B, initially referred to as LRP-DIT) was cloned and characterized as a candidate tumor suppressor. It is a new member of the low density lipoprotein receptor gene family. Its overall domain structure and large size (approximately 600 kDa) are similar to LRP and suggest that it is a multifunctional cell surface receptor. Herein, we characterize a series of ligands for the receptor using cell lines that stably express it as a domain IV minireceptor (mLRP1B4). Ligands of LRP including receptor-associated protein, urokinase plasminogen activator, tissue type plasminogen activator, and plasminogen activator inhibitor type-1 each demonstrate binding, internalization, and degradation via mLRP1B4. Interestingly, the kinetics of ligand endocytosis is distinctly different from that of LRP, with LRP1B exhibiting a markedly diminished internalization rate. In addition, tissue expression analysis reveals that the LRP1B gene is expressed in brain, thyroid, and salivary gland. These studies thus extend the physiological roles of members of the LDL receptor family. PMID- 11384979 TI - Zipper-interacting protein kinase induces Ca(2+)-free smooth muscle contraction via myosin light chain phosphorylation. AB - The inhibition of myosin phosphatase evokes smooth muscle contraction in the absence of Ca(2+), yet the underlying mechanisms are not understood. To this end, we have cloned smooth muscle zipper-interacting protein (ZIP) kinase cDNA. ZIP kinase is present in various smooth muscle tissues including arteries. Triton X 100 skinning did not diminish ZIP kinase content, suggesting that ZIP kinase associates with the filamentous component in smooth muscle. Smooth muscle ZIP kinase phosphorylated smooth muscle myosin as well as the isolated 20-kDa myosin light chain in a Ca(2+)/calmodulin-independent manner. ZIP kinase phosphorylated myosin light chain at both Ser(19) and Thr(18) residues with the same rate constant. The actin-activated ATPase activity of myosin increased significantly following ZIP kinase-induced phosphorylation. Introduction of ZIP kinase into Triton X-100-permeabilized rabbit mesenteric artery provoked a Ca(2+)-free contraction. A protein phosphatase inhibitor, microcystin LR, also induced contraction in the absence of Ca(2+), which was accompanied by an increase in both mono- and diphosphorylation of myosin light chain. The observed sensitivity of the microcystin-induced contraction to various protein kinase inhibitors was identical to the sensitivity of isolated ZIP kinase to these inhibitors. These results suggest that ZIP kinase is responsible for Ca(2+) independent myosin phosphorylation and contraction in smooth muscle. PMID- 11384980 TI - Identification of potential mechanisms for regulation of p115 RhoGEF through analysis of endogenous and mutant forms of the exchange factor. AB - Rho GTPases play a fundamental role in numerous cellular processes that are initiated by extracellular stimuli including agonists that work through G protein coupled receptors. A direct pathway for such regulation was elucidated by the identification of p115 RhoGEF, an exchange factor for RhoA that is activated through its RGS domain by G alpha(13). Endogenous p115 RhoGEF was found mainly in the cytosol of serum-starved cells but partially localized to membranes in cells stimulated with lysophosphatidic acid. Overexpressed p115 RhoGEF was equally distributed between membranes and cytosol; either the RGS or pleckstrin homology domain was sufficient for this partial targeting to membranes. Removal of the pleckstrin homology domain dramatically reduced the in vitro rate of p115 RhoGEF exchange activity. Deletion of amino acids 252--288 in the linker region between the RGS domain and the Dbl homology domain or of the last 150 C-terminal amino acids resulted in non-additive reduction of in vitro exchange activity. In contrast, p115 RhoGEF pieces lacking this extended C terminus were over 5-fold more active than the full-length exchange factor in vivo. These results suggest that p115 RhoGEF is inhibited in the cellular milieu through modification or interaction of inhibitory factors with its C terminus. Endogenous p115 RhoGEF that was immunoprecipitated from cells stimulated with lysophosphatidic acid or sphingosine 1-phosphate was more active than when the enzyme was immunoprecipitated from untreated cells. This indicates an additional and potentially novel long lived mechanism for regulation of p115 RhoGEF by G protein coupled receptors. PMID- 11384981 TI - Cloning of a mouse beta 1,3 N-acetylglucosaminyltransferase GlcNAc(beta 1,3)Gal(beta 1,4)Glc-ceramide synthase gene encoding the key regulator of lacto series glycolipid biosynthesis. AB - The distinction between the different classes of glycolipids is conditioned by the action of specific core transferases. The entry point for lacto-series glycolipids is catalyzed by the beta1,3 N-acetylglucosaminyltransferase GlcNAc(beta1,3)Gal(beta1,4)Glc-ceramide (Lc3) synthase enzyme. The Lc3 synthase activity has been shown to be regulated during development, especially during brain morphogenesis. Here, we report the molecular cloning of a mouse gene encoding an Lc3 synthase enzyme. The mouse cDNA included an open reading frame of 1131 base pairs encoding a protein of 376 amino acids. The Lc3 synthase protein shared several structural motifs previously identified in the members of the beta1,3 glycosyltransferase superfamily. The Lc3 synthase enzyme efficiently utilized the lactosyl ceramide glycolipid acceptor. The identity of the reaction products of Lc3 synthase-transfected CHOP2/1 cells was confirmed by thin-layer chromatography immunostaining using antibodies TE-8 and 1B2 that recognize Lc3 and Gal(beta1,4)GlcNAc(beta1,3)Gal(beta1,4)Glc-ceramide (nLc4) structures, respectively. In addition to the initiating activity for lacto-chain synthesis, the Lc3 synthase could extend the terminal N-acetyllactosamine unit of nLc4 and also had a broad specificity for gangliosides GA1, GM1, and GD1b to generate neolacto-ganglio hybrid structures. The mouse Lc3 synthase gene was mainly expressed during embryonic development. In situ hybridization analysis revealed that that the Lc3 synthase was expressed in most tissues at embryonic day 11 with elevated expression in the developing central nervous system. Postnatally, the expression was restricted to splenic B-cells, the placenta, and cerebellar Purkinje cells where it colocalized with HNK-1 reactivity. These data support a key role for the Lc3 synthase in regulating neolacto-series glycolipid synthesis during embryonic development. PMID- 11384982 TI - Mapping the epitope in cadherin-like receptors involved in Bacillus thuringiensis Cry1A toxin interaction using phage display. AB - In susceptible lepidopteran insects, aminopeptidase N and cadherin-like proteins are the putative receptors for Bacillus thuringiensis (Bt) toxins. Using phage display, we identified a key epitope that is involved in toxin-receptor interaction. Three different scFv molecules that bind Cry1Ab toxin were obtained, and these scFv proteins have different amino acid sequences in the complementary determinant region 3 (CDR3). Binding analysis of these scFv molecules to different members of the Cry1A toxin family and to Escherichia coli clones expressing different Cry1A toxin domains showed that the three selected scFv molecules recognized only domain II. Heterologous binding competition of Cry1Ab toxin to midgut membrane vesicles from susceptible Manduca sexta larvae using the selected scFv molecules showed that scFv73 competed with Cry1Ab binding to the receptor. The calculated binding affinities (K(d)) of scFv73 to Cry1Aa, Cry1Ab, and Cry1Ac toxins are in the range of 20-51 nm. Sequence analysis showed this scFv73 molecule has a CDR3 significantly homologous to a region present in the cadherin-like protein from M. sexta (Bt-R(1)), Bombyx mori (Bt-R(175)), and Lymantria dispar. We demonstrated that peptides of 8 amino acids corresponding to the CDR3 from scFv73 or to the corresponding regions of Bt-R(1) or Bt-R(175) are also able to compete with the binding of Cry1Ab and Cry1Aa toxins to the Bt-R(1) or Bt-R(175) receptors. Finally, we showed that synthetic peptides homologous to Bt-R(1) and scFv73 CDR3 and the scFv73 antibody decreased the in vivo toxicity of Cry1Ab to M. sexta larvae. These results show that we have identified the amino acid region of Bt-R(1) and Bt-R(175) involved in Cry1A toxin interaction. PMID- 11384983 TI - Physical interaction of CcmC with heme and the heme chaperone CcmE during cytochrome c maturation. AB - Biogenesis of c-type cytochromes requires the covalent attachment of heme to the apoprotein. In Escherichia coli, this process involves eight membrane proteins encoded by the ccmABCDEFGH operon. CcmE binds heme covalently and transfers it to apocytochromes c in the presence of other Ccm proteins. CcmC is necessary and sufficient to incorporate heme into CcmE. Here, we report that the CcmC protein directly interacts with heme. We further show that CcmC co-immunoprecipitates with CcmE. CcmC contains two conserved histidines and a signature sequence, the so-called tryptophan-rich motif, which is the only element common to cytochrome c maturation proteins of bacteria, archae, plant mitochondria, and chloroplasts. We report that mutational changes of these motifs affecting the function of CcmC in cytochrome c maturation do not influence heme binding of CcmC. However, the mutants are defective in the CcmC-CcmE interaction, suggesting that these motifs are involved in the formation of a CcmC-CcmE complex. We propose that CcmC, CcmE, and heme interact directly with each other, establishing a periplasmic heme delivery pathway for cytochrome c maturation. PMID- 11384984 TI - Muf1, a novel Elongin BC-interacting leucine-rich repeat protein that can assemble with Cul5 and Rbx1 to reconstitute a ubiquitin ligase. AB - The heterodimeric Elongin BC complex has been shown to interact in vitro and in mammalian cells with a conserved BC-box motif found in a growing number of proteins including RNA polymerase II elongation factor Elongin A, SOCS-box proteins, and the von Hippel-Lindau (VHL) tumor suppressor protein. Recently, the VHL-Elongin BC complex was found to interact with a module composed of Cullin family member Cul2 and RING-H2 finger protein Rbx1 to reconstitute a novel E3 ubiquitin ligase that activates ubiquitylation by the E2 ubiquitin-conjugating enzymes Ubc5 and Cdc34. In the context of the VHL ubiquitin ligase, Elongin BC functions as an adaptor that links the VHL protein to the Cul2/Rbx1 module, raising the possibility that the Elongin BC complex could function as an integral component of a larger family of E3 ubiquitin ligases by linking alternative BC box proteins to Cullin/Rbx1 modules. In this report, we describe identification and purification from rat liver of a novel leucine-rich repeat-containing BC-box protein, MUF1, which we demonstrate is capable of assembling with a Cullin/Rbx1 module containing the Cullin family member Cul5 to reconstitute ubiquitin ligase activity. In addition, we show that the additional BC-box proteins Elongin A, SOCS1, and WSB1 are also capable of assembling with the Cul5/Rbx1 module to reconstitute potential ubiquitin ligases. Taken together, our findings identify MUF1 as a new member of the BC-box family of proteins, and they predict the existence of a larger family of Elongin BC-based E3 ubiquitin ligases. PMID- 11384985 TI - Flagellar protein dynamics in Chlamydomonas. AB - Cilia and flagella appear to be stable, terminal, microtubule-containing organelles, but they also elongate and shorten in response to a variety of signals. To understand mechanisms that regulate flagellar dynamics, Chlamydomonas cells with nongrowing flagella were labeled with (35)S, and flagella and basal body components were examined for labeled polypeptides. Maximal incorporation of label into the flagella occurred within 3 h. Twenty percent of the flagellar polypeptides were exchanged. These included tubulins, dyneins, and 80 other axonemal and membrane plus matrix polypeptides. The most stable flagellar structure is the PF-ribbon, which comprises part of the wall of each doublet microtubule and is composed of tubulin and three other polypeptides. Most (35)S was incorporated into the high molecular weight ribbon polypeptide, rib240, and little, if any, (35)S is incorporated into PF-ribbon-associated tubulin. Both wild-type (9 + 2) and 9 + 0 flagella, which lack central microtubules, exhibited nearly identical exchange patterns, so labeling is not due to turnover of relatively labile central microtubules. To determine if flagellar length is balanced by protein exchange, (35)S incorporation into disassembling flagella was examined, as was exchange in flagella in which microtubule assembly was blocked by colchicine. Incorporation of (35)S-labeled polypeptides was found to occur into flagellar axonemes during wavelength-dependent shortening in pf18 and in fla10 cells induced to shorten flagella by incubation at 33 degrees C. Colchicine blocked tubulin addition but did not affect the exchange of the other exchangeable polypeptides; nor did it induce any change in flagellar length. Basal bodies also incorporated newly synthesized proteins. These data reveal that Chlamydomonas flagella are dynamic structures that incorporate new protein both during steady state and as flagella shorten and that protein exchange does not, alone, explain length regulation. PMID- 11384986 TI - Endophilin regulates JNK activation through its interaction with the germinal center kinase-like kinase. AB - The endophilin family of proteins function in clathrin-mediated endocytosis. Here, we have identified and cloned the rat germinal center kinase-like kinase (rGLK), a member of the GCK (germinal center kinase) family of c-Jun N-terminal kinase (JNK) activating enzymes, as a novel endophilin I-binding partner. The interaction occurs both in vitro and in cells and is mediated by the Src homology 3 domain of endophilin I and a region of rGLK containing the endophilin consensus binding sequence PPRPPPPR. Overlay analysis of rat brain extracts demonstrates that endophilin I is a major Src homology 3 domain-binding partner for rGLK. Overexpression of full-length endophilin I activates rGLK-mediated JNK activation, whereas N- and C-terminal fragments of endophilin I block JNK activation. Thus, endophilin I appears to have a novel function in JNK activation. PMID- 11384987 TI - Directing the immune response to carbohydrate antigens. AB - Peptide mimetics may substitute for carbohydrate antigens in vaccine design applications. At present, the structural and immunological aspects of antigenic mimicry, which translate into immunologic mimicry, as well as the functional correlates of each, are unknown. In contrast to screening peptide display libraries, we demonstrate the feasibility of a structure-assisted vaccine design approach to identify functional mimeotopes. By using concanavalin A (ConA), as a recognition template, peptide mimetics reactive with ConA were identified. Designed peptides were observed to compete with synthetic carbohydrate probes for ConA binding, as demonstrated by enzyme-linked immunosorbent assay and isothermal titration calorimetry (ITC) analysis. ITC measurements indicate that a multivalent form of one particular mimetic binds to ConA with similar affinity as does trimannoside. Splenocytes from mimeotope-immunized mice display a peptide specific cellular response, confirming a T-cell-dependent nature for the mimetic. As ConA binds to the Envelope protein of the human immunodeficiency virus, type 1 (HIV-1), we observed that mimeotope-induced serum also binds to HIV-1-infected cells, as assessed by flow cytometry, and could neutralize T-cell line adapted HIV-1 isolates in vitro, albeit at low titers. These studies emphasize that mimicry is based more upon functional rather than structural determinants that regulate mimeotope-induced T-dependent antibody responses to polysaccharide and emphasize that rational approaches can be employed to develop further vaccine candidates. PMID- 11384988 TI - Receptor proximity, not intermolecular orientation, is critical for triggering T cell activation. AB - Engagement of antigen receptors on the surface of T-cells with peptides bound to major histocompatibility complex (MHC) proteins triggers T-cell activation in a mechanism involving receptor oligomerization. Receptor dimerization by soluble MHC oligomers is sufficient to induce several characteristic activation processes in T-cells including internalization of engaged receptors and up-regulation of cell surface proteins. In this work, the influence of intermolecular orientation within the activating receptor dimer was studied. Dimers of class II MHC proteins coupled in a variety of orientations and topologies each were able to activate CD4+ T-cells, indicating that triggering was not dependent on a particular receptor orientation. In contrast to the minimal influence of receptor orientation, T-cell triggering was affected by the inter-molecular distance between MHC molecules, and MHC dimers coupled through shorter cross-linkers were consistently more potent than those coupled through longer cross-linkers. These results are consistent with a mechanism in which intermolecular receptor proximity, but not intermolecular orientation, is the key determinant for antigen induced CD4+ T-cell activation. PMID- 11384990 TI - The carboxyl-terminal valine is required for transport of glycoprotein CD8 alpha from the endoplasmic reticulum to the intermediate compartment. AB - There is evidence that a carboxyl-terminal valine residue is an anterograde transport signal for type I transmembrane proteins. Removal of the signal would either delay glycosylation in the Golgi complex of proteins destined to recycle to the endoplasmic reticulum or determine accumulation in the endoplasmic reticulum of newly synthesized proteins destined for the plasma membrane. We used the human CD8 alpha glycoprotein to investigate the role of the carboxyl-terminal valine in the exocytic pathway. Using immunofluorescence light microscopy, metabolic labeling, and cell fractionation, we demonstrate that removal of the carboxyl-terminal valine residue delays transport of CD8 alpha from the endoplasmic reticulum to the intermediate compartment. Removal of the residue did not affect the other steps of the exocytic pathway or the folding/dimerization and glycosylation processes. Therefore, it is likely that this signal plays a role in the transport of CD8 alpha from the endoplasmic reticulum to the intermediate compartment either before or during the formation of the transport vesicles that drive the exit the protein from the endoplasmic reticulum. PMID- 11384989 TI - Conditional expression of RNase P in the cyanobacterium Synechocystis sp. PCC6803 allows detection of precursor RNAs. Insight in the in vivo maturation pathway of transfer and other stable RNAs. AB - We have constructed a strain (CT1) that expresses RNase P conditionally with the aim to analyze the in vivo tRNA processing pathway and the biological role that RNase P plays in Synechocystis 6803. In this strain, the rnpB gene, coding for the RNA subunit of RNase P, has been placed under the control of the petJ gene promoter (P(petJ)), which is repressed by copper, cell growth, and accumulation of RNase P RNA is inhibited in CT1 after the addition of copper, indicating that the regulation by copper is maintained in the chimerical P(petJ)-rnpB gene and that RNase P is essential for growth in Synechocystis. We have analyzed several RNAs by Northern blot and primer extension in CT1. Upon addition of copper to the culture medium, precursors of the mature tRNAs are detected. Furthermore, our results indicate that there is a preferred order in the action of RNase P when it processes a dimeric tRNA precursor. The precursors detected are 3'-processed, indicating that 3' processing can occur before 5' processing by RNase P. The size of the precursors suggests that the terminal CCA sequence is already present before RNase P processing. We have also analyzed other potential RNase P substrates, such as the precursors of tmRNA and 4.5 S RNA. In both cases, accumulation of larger than mature size RNAs is observed after transferring the cells to a copper-containing medium. PMID- 11384991 TI - Repression of bacteriophage phi 29 early promoter C2 by viral protein p6 is due to impairment of closed complex. AB - Bacillus subtilis phage phi 29 encodes a very abundant protein, p6, which is a non sequence-specific DNA-binding protein. Protein p6 has the potential to bind cooperatively to the phage genome, forming a nucleoprotein complex in which the DNA adopts a right-handed toroidal conformation winding around a protein core. The formation of this complex at the right end of the phage genome where the early promoter C2 is located affects local topology, which may contribute to the promoter repression, although the underlying molecular mechanism of this repression is not presently known. In this study, we analyzed the effect of the p6 nucleoprotein complex on the formation of transcription complexes at the C2 promoter. The results obtained indicate that the nucleoprotein complex does not occlude promoter C2 to RNA polymerase because both proteins can bind to the same DNA molecule. Protein p6 binds along the fragment including the sequence adjacent to the bound polymerase, altering the structure of the transcriptional complex and affecting specifically the stability of the closed complex. The findings presented might help to answer some of the open questions about the concerted molecular mechanisms of histone-like proteins as transcriptional silencers. PMID- 11384992 TI - The Mdm-2 amino terminus is required for Mdm2 binding and SUMO-1 conjugation by the E2 SUMO-1 conjugating enzyme Ubc9. AB - Covalent attachment of SUMO-1 to Mdm2 requires the activation of a heterodimeric Aos1-Uba2 enzyme (ubiquitin-activating enzyme (E1)) followed by the conjugation of Sumo-1 to Mdm2 by Ubc9, a protein with a strong sequence similarity to ubiquitin carrier proteins (E2s). Upon Sumo-1 conjugation, Mdm2 is protected from self-ubiquitination and elicits greater ubiquitin-protein isopeptide ligase (E3) activity toward p53, thereby increasing its oncogenic potential. Because of the biological implication of Mdm2 sumoylation, we mapped Ubc9 binding on Mdm2. Here we demonstrate that Ubc9 can associate with Mdm2 only if amino acids 40-59 within the N terminus of Mdm2 are present. Mdm2 from which amino acids 40-59 have been deleted can no longer be sumoylated. Furthermore, addition of a peptide that corresponds to amino acids 40-59 on Mdm2 to a sumoylation reaction efficiently inhibits Mdm2 sumoylation in vitro and in vivo. In UV-treated cells Mdm2 exhibits reduced association with Ubc9, which coincides with decreased Mdm2 sumoylation. Our findings regarding the association of Ubc9 with Mdm2, and the effect of UV irradiation on Ubc9 binding, point to an additional level in the regulation of Mdm2 sumoylation under normal growth conditions as well as in response to stress conditions. PMID- 11384993 TI - Interaction between the noncatalytic region of Sid1p kinase and Cdc14p is required for full catalytic activity and localization of Sid1p. AB - Sid1p is a group II p21-activated kinase/germinal center kinase family member that is part of a signaling network required for cytokinesis in fission yeast. Germinal center kinases are characterized by well conserved amino-terminal catalytic domains followed by less conserved carboxyl termini. The carboxyl termini among group I germinal center kinases are moderately conserved and thought to be regulatory regions. Little is known about the carboxyl termini of group II family members. Sid1p has been shown to bind the novel protein Cdc14p; however, the functional significance of this interaction is unknown. Here we report that the carboxyl terminus of Sid1p is an essential regulatory region. Our results indicate that this region contains the binding domain for Cdc14p, and this association is required for full Sid1p catalytic activity as well as intracellular localization. Furthermore, overexpression of the carboxyl terminus of Sid1p alone compromises the signaling of cytokinesis. We conclude that Cdc14p positively regulates the Sid1p kinase by binding the noncatalytic carboxyl terminal region of the protein. PMID- 11384994 TI - Inhibition of LZIP-mediated transcription through direct interaction with a novel host cell factor-like protein. AB - Host cell factor 1 (HCF-1) is a cellular transcriptional coactivator which coordinates the assembly of enhancer complex through direct interactions with viral and cellular trans-activators such as VP16, Oct-1, LZIP, and GA-binding protein. These interactions are mediated by the beta-propeller domain comprising the first 380 residues of HCF-1 with six kelch repeats. Here we describe the identification and characterization of a novel HCF-like kelch repeat protein, designated HCLP-1. HCLP-1 is a ubiquitously expressed nuclear protein which is composed almost entirely of a six-bladed beta-propeller. HCLP-1 selectively interacts with LZIP but not with VP16. The physical interaction between HCLP-1 and LZIP leads to the repression of the LZIP-dependent transcription. The HCLP-1 binding domain of LZIP maps to residues 109-315, which contain the bZIP DNA binding motif. Electrophoretic mobility shift assay demonstrates that HCLP-1 indeed interferes with the binding of LZIP to its DNA target. Thus, HCLP-1 serves a transcriptional co-repressor function mediated through its inhibitory interaction with the LZIP transcription factor. Our findings suggest a new mechanism for transcriptional regulation by HCF-like proteins. PMID- 11384995 TI - Sp1 plays a critical role in the transcriptional activation of the human cyclin dependent kinase inhibitor p21(WAF1/Cip1) gene by the p53 tumor suppressor protein. AB - In the present study we present evidence for the critical role of Sp1 in the mechanism of transactivation of the human cell cycle inhibitor p21(WAF1/Cip1) (p21) gene promoter by the tumor suppressor p53 protein. We found that the distal p53-binding site of the p21 promoter acts as an enhancer on the homologous or heterologous promoters in hepatoma HepG2 cells. In transfection experiments, p53 transactivated the p21 promoter in HaCaT cells that express Sp1 but have a mutated p53 form. In contrast, p53 could not transactivate the p21 promoter in the Drosophila embryo-derived Schneider's SL2 cells that lack endogenous Sp1 or related factors. Cotransfection of SL2 cells with p53 and Sp1 resulted in a synergistic transactivation of the p21 promoter. Synergistic transactivation was greatly decreased in SL2 cells and HaCaT cells by mutations in either the p53 binding site or in the -82/-77 Sp1-binding site indicating functional cooperation between Sp1 and p53 in the transactivation of the p21 promoter. Synergistic transactivation was also decreased by mutations in the transactivation domain of p53. Physical interactions between Sp1 and p53 proteins were established by glutathione S-transferase pull-down and coimmunoprecipitation assays. By using deletion mutants we found that the DNA binding domain of Sp1 is required for its physical interaction with p53. In conclusion, Sp1 must play a critical role in regulating important biological processes controlled by p53 via p21 gene activation such as DNA repair, cell growth, differentiation, and apoptosis. PMID- 11384996 TI - Association of a novel PDZ domain-containing peripheral Golgi protein with the Q SNARE (Q-soluble N-ethylmaleimide-sensitive fusion protein (NSF) attachment protein receptor) protein syntaxin 6. AB - PDZ domains are involved in the scaffolding and assembly of multi-protein complexes at various subcellular sites. We describe here the isolation and characterization of a novel PDZ domain-containing protein that localizes to the Golgi apparatus. Using an in silico cloning approach, we have identified and isolated a cDNA encoding a ubiquitously expressed 59-kDa protein that we call FIG. It is composed of two coiled coil regions, a leucine zipper, and a single PDZ domain. Cytological studies using indirect immunofluorescence microscopy revealed that FIG is a peripheral protein that uses one of its coiled coil domains to localize to the Golgi apparatus. To ascertain the modalities of this Golgi localization, the same coiled coil region was tested for its ability to interact with a panel of coiled coil domain-containing integral membrane Golgi proteins. Using a series of GST fusion protein binding assays, co immunofluorescence and co-immunoprecipitation experiments, we show that FIG specifically binds to the coiled coil domain-containing Q-SNARE (Q-soluble NSF attachment protein receptor) protein syntaxin 6 both in vitro and in vivo. The structural features of FIG and its interaction with a SNARE protein suggest that FIG may play a role in membrane vesicle trafficking. This is the first example of a PDZ domain-containing peripheral protein that localizes to the Golgi through a coiled coil-mediated interaction with a resident membrane protein. Our results broaden the scope of PDZ domain-mediated functions. PMID- 11384998 TI - Structural imaging in the clinical diagnosis of Alzheimer's disease: problems and tools. PMID- 11384997 TI - A novel mechanism of carbohydrate recognition by the C-type lectins DC-SIGN and DC-SIGNR. Subunit organization and binding to multivalent ligands. AB - DC-SIGN and DC-SIGNR are cell-surface receptors that mediate cell-cell interactions within the immune system by binding to intercellular adhesion molecule-3. The receptor polypeptides share 77% amino acid sequence identity and are type II transmembrane proteins. The extracellular domain of each comprises seven 23-residue tandem repeats and a C-terminal C-type carbohydrate-recognition domain (CRD). Cross-linking, equilibrium ultracentrifugation, and circular dichroism studies of soluble recombinant fragments of DC-SIGN and DC-SIGNR have been used to show that the extracellular domain of each receptor is a tetramer stabilized by an alpha-helical stalk. Both DC-SIGN and DC-SIGNR bind ligands bearing mannose and related sugars through the CRDs. The CRDs of DC-SIGN and DC SIGNR bind Man(9)GlcNAc(2) oligosaccharide 130- and 17-fold more tightly than mannose, and affinity for a glycopeptide bearing two such oligosaccharides is increased by a further factor of 5- to 25-fold. These results indicate that the CRDs contain extended or secondary oligosaccharide binding sites that accommodate mammalian-type glycan structures. When the CRDs are clustered in the tetrameric extracellular domain, their arrangement provides a means of amplifying specificity for multiple glycans on host molecules targeted by DC-SIGN and DC SIGNR. Binding to clustered oligosaccharides may also explain the interaction of these receptors with the gp120 envelope protein of human immunodeficiency virus 1, which contributes to virus infection. PMID- 11385000 TI - Visual hallucinations in Parkinson's disease: their nature, frequency, and origins. PMID- 11385001 TI - Anosmia in dementia is associated with Lewy bodies rather than Alzheimer's pathology. PMID- 11385002 TI - Imaging counterparts of cognitive decline in multiple sclerosis. PMID- 11385003 TI - The late whiplash syndrome: a biopsychosocial approach. AB - Physicians and other therapists continue to grapple in daily practice with the controversies of the late whiplash syndrome. For decades much of the debate and the approach to this controversial syndrome has centred on the natural history of and progression to chronic pain after acute whiplash injury. Recognising that there is recent epidemiological data that defines the natural history of the acute whiplash injury outside of many of the confounding factors occurring in many western countries, and the lack of evidence for a "chronic whiplash injury", this article will thus introduce the biopsychosocial model, its elements, its advantages over the traditional model, and the practical application of this model. The biopsychosocial model recognises physical and psychological sources of somatic symptoms, but fundamentally recognises that the late whiplash syndrome is not the result of a "chronic injury". PMID- 11385004 TI - Visual hallucinations in Parkinson's disease: a review and phenomenological survey. AB - OBJECTIVES: Between 8% and 40% of patients with Parkinson's disease undergoing long term treatment will have visual hallucinations during the course of their illness. There were two main objectives: firstly, to review the literature on Parkinson's disease and summarise those factors most often associated with hallucinations; secondly, to carry out a clinical comparison of ambulant patients with Parkinson's disease with and without visual hallucinations, and provide a detailed phenomenological analysis of the hallucinations. METHODS: A systematic literature search using standard electronic databases of published surveys and case-control studies was undertaken. In parallel, a two stage questionnaire survey was carried out based on members of a local branch of the Parkinson's Disease Society and followed up with a clinical interview. RESULTS: The review disclosed common factors associated with visual hallucinations in Parkinson's disease including greater age and duration of illness, cognitive impairment, and depression and sleep disturbances. The survey comprised 21 patients with visual hallucinations and 23 without. The hallucinators had a longer duration and a greater severity of illness, and tended to show more depressed mood and cognitive impairment. The typical visual hallucination in these patients is a complex visual image experienced while they are alert and have their eyes open. The image appears without any known trigger or voluntary effort, is somewhat blurred, and commonly moves. It stays present for a period of "seconds" or "minutes". The content can be variable within and between hallucinators, and includes such entities as people, animals, buildings, or scenery. These features resemble those highlighted in hallucinations in the visually impaired (Charles Bonnet's syndrome). CONCLUSION: A consistent set of factors are associated with visual hallucinations in Parkinson's disease. The results of the phenomenological survey and those of visual hallucinations carried out in other settings suggest a common physiological substrate for visual hallucinations but with cognitive factors playing an as yet unspecified role. PMID- 11385005 TI - Prospective study of hallucinations and delusions in Parkinson's disease. AB - OBJECTIVE: This study was undertaken to determine the prevalence of hallucinations and delusions in Parkinson's disease, to describe such symptoms phenomenologically, and possibly to determine factors associated with their development. In addition, the role of the visual system in relation to visual hallucinations was examined. METHOD: 102 consecutive patients diagnosed with strictly defined Parkinson's disease were examined for the presence of hallucinations and delusions and assessed for visual acuity, cognition, depression using the geriatric depression scale, disease severity as measured by the UPDRS, and other clinical variables. RESULTS: Of 102 consecutive patients, 29.4% (n=30) had hallucinations or delusions, four (3.9%) were determined to be psychotic due solely to delirium and were excluded from further analysis. Of the 98 remaining patients, 26.5% (n=26) had visual hallucinations. Among these, one patient also had delusions, two had auditory hallucinations, and one had gustatory hallucinations. Visual hallucinations were significantly associated with worse visual acuity, lower cognitive score, higher depression score, and worse disease severity. Hallucinations were not associated with history of psychiatric disease, dose or duration of levodopa or other antiparkinsonian medication treatment, or duration of illness. CONCLUSIONS: Visual hallucinations are common symptoms in Parkinson's disease and are most likely of multifactorial origin. Although higher doses of levodopa are known to be related clinically to hallucinations in individual patients, the results suggest that several underlying characteristics of patients with Parkinson's disease (disease severity, dementia, depression, worse visual acuity) may be more important determinants of which patients experience hallucinations. The data also provide preliminary evidence that abnormality of the visual system may be related to visual hallucinations in Parkinson's disease, as has been found in other disorders with visual hallucinations. PMID- 11385006 TI - Anosmia in dementia is associated with Lewy bodies rather than Alzheimer's pathology. AB - OBJECTIVES: To assess olfactory function of patients with dementia. Odour detection ability is impaired in clinical Parkinson's disease. Evidence of impaired detection in patients with clinically diagnosed Alzheimer's disease is inconsistent. No studies of olfaction have been neuropathologically validated. METHODS: The olfactory function of 92 patients with dementia and 94 controls was assessed using a simple bedside test as part of the Oxford Project To Investigate Memory and Ageing (OPTIMA). Neuropathological assessment was made of cortical Lewy bodies and substantia nigra (SN) cell counts and of Alzheimer's disease in all 92 patients, 22 of whom had SN Lewy bodies and 43 of whom had only Alzheimer's disease. RESULTS: Patients with Lewy bodies were more likely to be anosmic than those with Alzheimer's disease or controls. Patients with Alzheimer's disease were not more likely to be anosmic than controls. Nor was anosmia associated with degree of neurofibrillary tangles, as assessed by Braak stage. Among subjects with Lewy bodies, overall cortical Lewy body scores and Lewy body density in the cingulate were higher in those who were anosmic. Consensus clinical criteria for dementia with Lewy bodies had a sensitivity of 64% and specificity of 89%. In the absence of definite Alzheimer's disease, the criteria had sensitivity of 100%. In patients with definite Alzheimer's disease, anosmia was slightly more sensitive (55%) than the consensus criteria (33%). However, the addition of anosmia to the consensus criteria did not improve their overall performance. CONCLUSION: Dementia with Lewy bodies is associated with impaired odour detection. Misdiagnosis may have accounted for some previous reports of impaired odour detection in Alzheimer's disease. Simple but more sensitive tests of anosmia are required if they are to be clinically useful in identifying patients with dementia with Lewy bodies. PMID- 11385007 TI - Vladimir Mikhailovich Bechterev (1857-1927). PMID- 11385008 TI - Use of 14-3-3 and other brain-specific proteins in CSF in the diagnosis of variant Creutzfeldt-Jakob disease. AB - OBJECTIVES: The detection of the protein 14-3-3 in the CSF has been shown to be a reliable and sensitive marker for sporadic Creutzfeldt-Jakob disease (CJD). Other brain-specific proteins such as neuron specific enolase (NSE), S-100b, and tau protein have also been reported to be increased in the CSF of patients with sporadic CJD. In 1996 a variant of CJD (vCJD) was described which is likely to be causally linked to the bovine spongiform encephalopathy agent. This study reports and compares the findings of CSF brain specific protein analysis in 45 patients with vCJD and in 34 control patients. METHODS: The CSF from 45 patients with vCJD and 34 controls were investigated for the presence of 14-3-3 by SDS polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting with chemiluminescent detection. Tau protein, S-100b, and NSE concentrations in CSF were measured using enzyme immunoassays. RESULTS: Protein 14-3-3 was detected in the CSF of 22/45 patients with vCJD and in 3/34 controls. The mean concentrations of NSE, S-100b, and tau protein in CSF were significantly raised in patients with vCJD compared with controls. The positive predictive value of CSF 14-3-3 was 86% and the negative predictive value was 63%. These values are lower than those reported for sporadic CJD. An increased CSF tau had a positive predictive value of 93% and a negative predictive value of 81%. The combination of CSF 14-3-3 and/or increased CSF tau had a positive predictive value of 91% and a negative predictive value of 84%. CONCLUSIONS: CSF protein 14-3-3 is not as useful a marker for vCJD as it is for sporadic CJD. Increased concentration of CSF tau was found to be a sensitive marker of vCJD but as concentrations may be increased in many forms of non-CJD dementia, this may limit its usefulness as a diagnostic test. PMID- 11385010 TI - Return to driving after head injury. AB - OBJECTIVES: To determine whether patients who return to driving after head injury can be considered safe to do so and to compare the patient characteristics of those who return to driving with those who do not. METHODS: In a multicentre qualitative study 10 rehabilitation units collectively registered 563 adults with traumatic brain injury during a 2.5 year period. Recruitment to the study varied from immediately after hospital admission to several years after injury. Patients and their families were interviewed around 3 to 6 months after recruitment. A total of 383 (67.5%) subjects were interviewed within 1 year of injury, of whom 270 (47.6%) were interviewed within 6 months of injury. Main outcome measures were the presence or absence of driving related problems reported by drivers and ex-drivers, and scores on driving related items of the functional independence/functional assessment measure (FIM+FAM). RESULTS: Of the 563 patients 381 were drivers before the injury and 139 had returned to driving at interview. Many current drivers reported problems with behaviour (anger, aggression, irritability; 67 (48.2%)), memory ( 89 (64%)), concentration and attention (39 (28.1%)), and vision (39 (28.1%)). Drivers reported most driving related problems as often as ex-drivers, main exceptions were epilepsy and community mobility. Current drivers scored significantly higher on the FIM+FAM (were more independent), than ex-drivers. The driving group had sustained less severe head injuries than ex-drivers; nevertheless, 78 (56.2%) current drivers had received a severe head injury. Few (61 (16%)) previous drivers reported receiving formal advice about driving after injury. CONCLUSIONS: The existence of problems which could significantly affect driving does not prevent patients returning to driving after traumatic brain injury. Patients should be assessed for both mental and physical status before returning to driving after a head injury, and systems put in place to enable clear and consistent advice to be given to patients about driving. PMID- 11385009 TI - Functional MRI for presurgical planning: problems, artefacts, and solution strategies. AB - OBJECTIVES: Presurgical mapping of motor function is a widely used clinical application of functional (f) MRI, employing the blood oxygenation level dependent contrast. The aim of this study was to report on 3 years experience of 194 fMRI studies on the representation of motor function in 103 patients and to describe the problems and artefacts that were typically present. METHODS: An evaluation was carried out to determine whether the patients' age, type or location of the tumourous lesion, severity of the paresis, or the tasks used during the investigation have an effect on artefacts of fMRI studies and how these artefacts are best overcome. RESULTS: Functional MRI identified the motor regions in 85% of all investigated paradigms. In 11% of the investigated patients no information at all on functional localisation was obtained. A draining vein within the central sulcus was present in all patients that showed activation within the parenchyma of the precentral gyrus but also in three patients in whom no parenchymal activation was present. Head movement artefacts were the most frequent cause for fMRI failure, followed by low signal to noise ratio. Motion artefacts were correlated with the degree of paresis and with the functional task. Tasks involving more proximal muscles led to significantly more motion artefacts when compared with tasks that primarily involved distal muscles. Mean MR signal change during task performance was 2.5%. CONCLUSIONS: Most of the artefacts of functional MRI can be reliably detected and at least in part be reduced or eliminated with the help of mathematical algorithms, appropriate pulse sequences and tasks, and-probably most important-by evaluating the fMRI raw data that is, the MR signal time courses. PMID- 11385011 TI - Oligoclonal T cell repertoire in cerebrospinal fluid of patients with inflammatory diseases of the nervous system. AB - OBJECTIVE: To evaluate the T cell receptor beta chain variable region (TCRBV) gene usage ex vivo in CSF cells and peripheral blood mononuclear cells (PBMCs) collected from patients with autoimmune and inflammatory diseases of the nervous system. METHODS: A novel sensitive seminestedpolymerase chain reaction coupled with heteroduplex analysis was developed. RESULTS: Under these experimental conditions, the minimal number of cells required for the analysis of the whole T cell repertoire was established at 2.5x10(4)-sufficient to evaluate most of the samples collected during diagnostic lumbar punctures. In the 21 patients examined, restrictions in TCRBV gene family usage were not seen. However, using heteroduplex analysis, oligoclonal T cell expansions were found in the CSF of 13 patients and monoclonal expansions in five patients. The T cell abnormalities found did not correlate with intrathecal IgG production or with any clinical variable considered. CONCLUSION: T cell clonal expansions, useful for further characterisation of pathogenetic T cells, can be found during the course of nervous system inflammations, but this abnormality is probably not disease specific. PMID- 11385012 TI - A longitudinal study of brain atrophy and cognitive disturbances in the early phase of relapsing-remitting multiple sclerosis. AB - OBJECTIVE: (a) To establish whether the cognitive decline of the early phase of relapsing-remitting multiple sclerosis depends on the progression of the burden of disease, or on the loss of brain parenchyma, or is influenced by both; (b) to monitor the loss of brain parenchyma in the early phase of the disease; and (c) to examine its possible relation with the progression of physical disability. METHODS: For 2 years 53 patients with clinically definite relapsing-remitting multiple sclerosis with disease duration 1-5 years and expanded disability status scale < or =5.0 at baseline were monitored. The neuropsychological performances, the psychological functioning, the neurological impairment, and the disability have been assessed at baseline and after 2 years. Patients also underwent PD/T2 and T1 weighted brain MRI. T2 and T1 lesion volumes were measured by a semiautomatic technique. Quantification of brain parenchymal volumes was obtained using a highly reproducible computerised interactive program. The relation between cognitive impairment and MRI findings has been investigated by partial correlation and stepwise multiple regression analyses excluding the effects of age, education, anxiety, depression, and total days of steroid use. RESULTS: In the 2 years of the study the mean change for T2 and T1 lesion volumes and brain parenchymal volumes were +1.7 ml (95% confidence interval (95% CI) 1.3-2.2, p=0.005, (29.8%); +0.2 ml, 95% CI 0.15-0.26, p=0.004, (25%); and -32.3 ml, 95% CI 24.2-42.3, p<0.0001, (2.7%), respectively. Overall, 14 patients (26.4%) were judged to be cognitively impaired at baseline and 28 (52.8%) at the end of the follow up. Of the 18 neuropsychological tests and subtests employed in the study, patients with multiple sclerosis failed 5.8 (SD 2.3) tests at the baseline and 8.4 (SD 2.9) (p<0.0001) tests at the end of the study. When the cognitive changes were examined in individual patients, five (9.4%) of them were considered cognitively improved, 33 (62.3%) remained stable, and 15 (28.3%) worsened over 2 years. T2 and T1 volume changes in improved, stable, and worsened patients did not show any significant difference, whereas brain parenchymal volume decrease in cognitively worsened patients was significantly greater (-66 ml (5.4%), 95% CI 37 108.9, p=0.0031). The cognitive impairment was independently predicted over 2 years only by the change of brain parenchymal volumes (R=0.51, p=0.0003). Ten patients (18.9%), who worsened by one or more points in the EDSS during the follow up period had significant decreases in brain parenchymal volumes (-99 ml (8%), 95% CI 47.6-182.3, p=0.005). At the end of the study the loss of brain parenchyma correlated significantly with change in EDSS (r= 0.59, p<0.0001). CONCLUSIONS: In the early phase of relapsing-remitting multiple sclerosis the cognitive deterioration relies more on the development of brain parenchymal volume atrophy than on the extent of burden of disease in the brain. The loss of brain parenchymal volume underlies the progressive accumulation of physical disability from the initial phase of the disease, which becomes more demonstrable only if studied with longer observation periods. Probably, the main pathological substrate of brain atrophy in the early stage of the disease is early axonal loss, which causes the progression of neurological deficits and the development of cognitive impairment. These data support the debated opinion that disease modifying therapy should be initiated as early as possible. PMID- 11385013 TI - Cardiac (123)I-meta-iodobenzylguanidine (MIBG) uptake in dementia with Lewy bodies: comparison with Alzheimer's disease. AB - Cardiac (123)I-meta-iodobenzylguanidine (MIBG) uptake was measured in 11 patients with dementia with Lewy bodies (DLB), 10 patients with Alzheimer's disease (AD), and 10 age matched control subjects. The severity of cognitive impairment and duration of symptoms in patients with DLB matched that in the patients with AD. The heart/mediastinum (H/M) ratio of MIBG uptake in the patients with AD was indistinguishable from that in the control subjects. However, the H/M ratio in all patients with DLB was significantly lower than that in the patients with AD and control subjects (p<0.001). These findings indicate that local myocardial sympathetic nerves are affected in DLB and that cardiac (123)I-MIBG scintigraphy may provide a means of differentiating DLB from AD. PMID- 11385014 TI - Increased plasma neurotensin concentrations in patients with Parkinson's disease. AB - Plasma neurotensin (NT) was measured by radioimmunoassay in propanol extracted and unextracted plasma from 16 parkinsonian patients (four before treatment) and 16 age and sex matched controls. Mean plasma NT concentrations were consistently higher in parkinsonian patients than in controls and higher in the four untreated patients than in levodopa treated patients suggesting that plasma NT measurement may represent an easy detectable additional index in diagnosing parkinsonism and provides a novel approach to research in this field. PMID- 11385015 TI - Visual field loss associated with vigabatrin: pathological correlations. AB - Pathological changes are reported in the anterior visual pathways of a 41 year old man with complex partial seizures treated with vigabatrin who developed bilateral visual field constriction. There was peripheral retinal atrophy with loss of ganglion cells and loss of nerve fibres in the optic nerves, chiasm, and tracts. No evidence of intramyelinic oedema was seen. These findings suggest that the primary site of injury lies within the ganglion cells in the retina. The degree of atrophy seen would suggest that the visual field loss is irreversible. PMID- 11385016 TI - Paroxysmal alternating skew deviation and nystagmus after partial destruction of the uvula. AB - A patient with suspected brain stem glioma involving the area of the left vestibular nuclei and cerebellar peduncle, developed paroxysmal alternating skew deviation and direction changing nystagmus after biopsy of the inferior cerebellar vermis resulting in destruction of the uvula. Between attacks she had right over left skew deviation with asymptomatic right beating horizontal nystagmus. Slow phases of the resting nystagmus showed increasing velocity, similar to congenital nystagmus. At intervals of 40-50 seconds, paroxysmal reversal of her skew deviation occurred, accompanied by violent left beating horizontal torsional nystagmus lasting 10-12 seconds and causing severe oscillopsia. It is proposed that this complex paroxysmal eye movement disorder results from (1) a lesion in the left vestibular nuclei causing right over left skew and right beating resting nystagmus and (2) a disruption of cerebellar inhibition of vestibular nuclei, causing alternating activity in the vestibular system with intermittent reversal of the skew deviation and paroxysmal nystagmus towards the side of the lesion. PMID- 11385017 TI - Unusual presentation of a primary spinal Burkitt's lymphoma. AB - Primary CNS lymphomas are detected with increasing frequency in immunocompetent and immunodeficient persons. Primary involvement of the spinal roots has only rarely been reported. The unusual history is described of a patient with a primary spinal Burkitt's lymphoma initially presenting as an S1 syndrome showing lymphocytic pleocytosis in the CSF, leading to the misdiagnosis of meningoradiculitis. Repeated spinal MRI disclosed a spinal mass lesion and histological and immunohistological examination of the tumour confirmed the diagnosis of spinal Burkitt's lymphoma. PMID- 11385018 TI - Juvenile muscular atrophy of the distal upper limb (Hirayama disease) associated with atopy. AB - Juvenile muscular atrophy of the distal upper limb (Hirayama disease) is a rare disease predominantly affecting the anterior horn cells of the cervical spinal cord in young men. Although the disease is considered to be a type of cervical myelopathy, the mechanism remains unknown. An immunological study of five consecutive patients with this disorder who were examined in the neurology clinic at Kyushu University Hospital during the past 2 years were performed. All developed distal muscular atrophy and weakness of one or both upper limbs in the second decade, and showed forward displacement of the dural sac and passive dilatation of the posterior venus plexus at the lower cervical portion on MRI during neck flexion. Four of the five patients had one or more coexistent airway allergies, such as allergic rhinitis, pollinosis, and asthma, and all five patients had a family history of atopic or allergic disorders in close relatives. Four of the five patients had mild eosinophilia. All five patients commonly had IgE specific to two mite antigens, Dermatophagoides pteronyssinus and Dermatophagoides farinae, whereas three of them also showed a raised total serum IgE concentration. The frequency of mite antigen specific IgE was significantly higher in the present patients with Hirayama disease than in 82 healthy controls (26/82, p<0.005). These findings suggest that atopy may be one of the contributing factors for Hirayama disease. PMID- 11385019 TI - Aphasia or aphemia. PMID- 11385020 TI - Inherited prion disease with A117V mutation of the prion protein gene: a novel Hungarian family. AB - Three members of a family with inherited prion disease are reported. One additional family member had a progressive neurological disease without details. Two developed symptoms of ataxia, dementia, myoclonus, rigidity, and hemiparesis, and one had a different phenotype with the combination of lower motor neuron deficit, parkinsonism, intellectual decline, and ataxia. In this last patient cell loss of the anterior horn motor neurons and chronic neurogenic muscle atrophy was evident. Immunostaining for the prion protein disclosed unicentric and multicentric plaques, and coarse and fine granular positivity. Genetic analysis of the prion protein gene of the propositus showed a 117 codon alanine to valine mutation and homozygous 129 valine/valine genotype. PMID- 11385021 TI - Transient total tongue paralysis from simultaneous central and peripheral lesions. PMID- 11385022 TI - Eye movement abnormality associated with cyclosporin. PMID- 11385023 TI - Preoperative and postoperative imaging of an aneurysm of the middle cerebral artery with transcranial duplex sonography. PMID- 11385042 TI - Neurology in practice: epilepsy. PMID- 11385043 TI - Assessment and investigation of possible epileptic seizures. PMID- 11385044 TI - If it's not epilepsy... PMID- 11385045 TI - The management of epilepsy. PMID- 11385046 TI - The management of status epilepticus. PMID- 11385047 TI - The discovery of the visual function of vitamin A. PMID- 11385048 TI - Jean Mayer 1920-1993. PMID- 11385049 TI - Casein phosphopeptides influence calcium uptake by cultured human intestinal HT 29 tumor cells. AB - We investigated the direct effects of casein phosphopeptides (CPP), which are formed by the proteolytic degradation of alpha- and beta-caseins, on calcium uptake by human HT-29 intestinal tumor cells, which undergo an enterocytically oriented differentiation in culture. A commercial preparation containing a mixture of purified CPP and an individual CPP of 25 amino acids, both containing the characteristic Ca(2+) binding motif, ser(P)-ser(P)-ser(P)-glu-glu, were employed. The study was performed at the single-cell level and on a cell population and measured the changes in cytosolic calcium concentration before and after CPP addition. In the presence of 2 mmol/L extracellular calcium, both CPP preparations induced a transient rise of free intracellular calcium ions, which did not influence ATP-induced release of calcium from intracellular stores, and which disappeared completely in the absence of extracellular calcium. Pretreatment of these cells with thapsigargin, which completely empties the intracellular calcium stores, did not abolish the cell responses to CPP. Repetitive stimulation of HT-29 cells with CPP always elicited a transient calcium rise, suggesting a lack of desensitization. The CPP-stimulated cytosolic calcium rise was dependent on CPP dose, in a seemingly nonsaturating mode, and on cell numbers. All of this is consistent with the hypothesis that CPP do not influence membrane-bound receptors or ion channels, but may act as calcium ionophores or calcium carriers across the membrane. The reported findings provide a new basis on which to assess the possibility that CPP enhance calcium absorption and bioavailability in animals. PMID- 11385050 TI - Garlic and garlic-derived compounds inhibit human squalene monooxygenase. AB - Although extracts of garlic inhibit cholesterol biosynthesis in cultured hepatocytes, the inhibitory components of garlic and the site or sites of inhibition in the cholesterol biosynthetic pathway have not been established. To elucidate potential mechanisms of inhibition, we examined the effect of fresh garlic extract and 16 water- or lipid-soluble compounds derived from garlic on purified recombinant human squalene monooxygenase. Squalene monooxygenase catalyzes the second and likely rate-limiting step in the downstream pathway for cholesterol biosynthesis. A 50% inhibitory concentration (IC(50)) of squalene epoxidation was achieved with 1 g/L of fresh garlic extract; of the 16 garlic compounds tested, only selenocystine (IC(50) = 65 micromol/L), S-allylcysteine (IC(50) = 110 micromol/L), alliin (IC(50) = 120 micromol/L), diallyl trisulfide (IC(50) = 195 micromol/L), and diallyl disulfide (IC(50) = 400 micromol/L) substantially inhibited the enzyme. Kinetic analysis showed that the inhibition by garlic and by these compounds was slow and irreversible, suggestive of covalent binding to the enzyme; the ability of thiol-containing compounds such as glutathione and 2,3-dimercaptopropanol to prevent and reverse the inhibition indicated that the garlic compounds were reacting with sulfhydryl groups on the protein. Dithiols were better reversal agents than monothiols, further suggesting that these inhibitors bind to the proposed vicinal sulfhydryls present on this enzyme. These results indicate that squalene monooxygenase may be one of the target enzymes through which garlic inhibits cholesterol biosynthesis. PMID- 11385051 TI - Dietary conjugated linoleic acid reduces adiposity in lean but not obese Zucker rats. AB - Recent studies have demonstrated a reduction in body fat in growing animals fed conjugated linoleic acid (CLA). Two experiments were conducted to extend these observations to obese rats so that the mechanism of the actions of CLA might be more easily elucidated. In experiment 1, male lean and obese Zucker rats were fed diets containing either 0 or 0.5% CLA for 5 wk. There was no effect of diet on growth rate or food intake. Dietary CLA reduced retroperitoneal and inguinal fat pad weights in the lean rats but increased fat pad weights in the obese genotype (diet x genotype interaction; P < 0.05). Determination of fat pad cellularity indicated that these changes in fat pad weight were due to a reduction or increase in average fat cell size for the lean and obese Zucker rats, respectively. In experiment 2, we sought to reproduce these effects on fat pad size, as well as to determine the effect of dietary CLA on the catabolic response to bacterial endotoxin injection in obese Zucker rats. Growing female lean and obese Zucker rats were fed diets containing 0 or 0.5% CLA for 8 wk. On d 28, each rat was injected intraperitoneally with lipopolysaccharide from Escherichia coli serotype 055:B5 (1 mg/kg body weight) and body weight was determined over the next 96 h. There was a diet x genotype interaction (P < 0.05) for the body weight response to lipopolysaccharide 24 h postinjection. Lean rats fed CLA lost less weight than did lean controls, but obese rats fed CLA lost more weight than did obese controls. As in the first experiment, there was a diet x genotype (P < 0.05) for the effect of treatment on retroperitoneal fat pad weights determined at the end of the experiment. Lean rats fed CLA had smaller RP fat pads than did lean controls, but obese rats fed CLA once again had heavier RP fat pads than did obese controls. These results indicate that CLA reduces body fat and catabolic response to endotoxin injection in lean Zucker rats but not in the obese genotype. The observed interaction between diet and genotype warrants additional investigation into the specific mechanism(s) of the biological activities of CLA. PMID- 11385052 TI - Dietary gamma-linolenic acid suppresses aortic smooth muscle cell proliferation and modifies atherosclerotic lesions in apolipoprotein E knockout mice. AB - The present study was conducted to evaluate the antiatherogenic effects of dietary gamma-linolenic acid (GLA) (primrose oil) in apolipoprotein E (apoE) genetic knockout mice. Five-wk-old male mice were fed cholesterol-free diets containing 10 g/100 g lipid as corn oil (CO) [control diet, 0 mol/100 mol GLA and (n-3) polyunsaturated fatty acids (PUFA)], primrose oil (PO, 10 mol/100 mol GLA), fish oil-CO mix [FC; 9:1 wt/wt, 0 mol/100 mol GLA and 17 mol/100 mol (n-3) PUFA] or fish oil-PO mix [FP, 1:3 wt/wt, 8 mol/100 mol GLA and 5 mol/100 mol (n-3) PUFA] for 15 wk. Subsequently, diets were supplemented with cholesterol (1.25 g/100 g) and sodium cholate (0.5 g/100 g) and fed for an additional 10 and 16 wk. Plasma cholesterol and triglyceride levels generally did not differ among groups at 20, 30 and 36 wk of age. Mice fed GLA-containing diets (PO and FP) had significantly (P < 0.05) higher liver phospholipid levels of dihomo-gamma linolenic acid, the elongated product of GLA, relative to CO and FC groups. Consumption of GLA (PO and FP diets) significantly reduced (P < 0.05) aortic vessel wall medial layer thickness at 20 and 30 wk. A parallel GLA-dependent suppression in the number of proliferating (proliferating cell nuclear antigen positive) aortic smooth muscle cells was also observed. Diets containing either GLA or (n-3) PUFA reduced (P < 0.05) atherosclerotic lesion size in 30-wk-old mice. These results indicate that dietary GLA can suppress smooth muscle cell proliferation in vivo and retard the development of diet-induced atherosclerosis in apoE knockout mice. PMID- 11385053 TI - Protein expressions of branched-chain keto acid dehydrogenase subunits are selectively and posttranscriptionally altered in liver and skeletal muscle of starved rats. AB - Although it has been well established that starvation increases the oxidation of branched-chain keto acids (BCKA) in humans and experimental animals such as rats, the mechanism has not been adequately investigated. For example, the effects of starvation on protein and mRNA expressions of BCKA dehydrogenase, which is the key enzyme regulating this oxidation, have not yet been studied. To initiate such studies, we first determined the activity of BCKA dehydrogenase in the liver and skeletal muscle of fed and starved rats. The levels of activity of BCKA dehydrogenase were significantly greater in tissues of starved rats than in those of fed rats. We then investigated the possible mechanisms of these increases in enzyme activity. The activity state of the enzyme was greater by 3-fold in the muscle of starved compared with fed rats, but there was no significant difference between the activity states in the liver. There were no significant differences between protein expressions of BCKA dehydrogenase subunits (E(1)alpha, E(1)beta and E(2)) in tissues of fed and starved rats; the exceptions were a greater expression of E(1)alpha in the liver and a lower expression of E(1)beta in the skeletal muscle of starved rats. These differences in protein expressions were not accompanied with any difference in the mRNA expressions of genes encoding E(1)alpha and E(1)beta. The rate of inactivation of BCKA dehydrogenase, mediated by its associated kinase, was significantly slower in the skeletal muscle of starved rats but was the same in the liver. However, there was no significant difference between the protein or the mRNA expressions of the gene encoding BCKA dehydrogenase kinase in tissues of fed and starved rats. These results show that starvation increases the activity of BCKA dehydrogenase in the liver and skeletal muscle, and the mechanisms of increases in activity are posttranscriptional and involve cellular rather than the molecular mechanisms. PMID- 11385054 TI - Food restriction differentially affects pituitary hormone mRNAs throughout the adult life span of male F344 rats. AB - Because neuroendocrine mechanisms may contribute to the antiaging effects of food restriction (FR), we measured the effect of FR on mRNAs encoding anterior pituitary (AP) tropic hormones. Slot blots or RNase protection assays were done on AP RNA from 3-, 6-, 12-, 18- and 24-mo-old male F344 rats consuming food ad libitum (AL) or food restricted (FR; to 60% of AL food intake) from 6 wk. Both AL and FR rats gained body weight during the study (P < 0.05), but FR rats weighed approximately 40% less (P < 0.0001). Messenger RNA levels were expressed in two ways, i.e., per total AP and per microgram total AP RNA. Proopiomelanocortin (POMC) mRNA/microg RNA was higher (P < 0.0005) in FR than in AL rats at all ages. Thyroid-stimulating hormone (TSH) beta mRNA declined with age (P < 0.05) in AL but not FR rats and was reduced by FR up to 12 mo (P < 0.01). Growth hormone (GH) mRNA/microg RNA declined with age (P < 0.05) in AL but not FR rats, and total GH mRNA in the AP was reduced by FR at early ages (P < 0.05). FR reduced prolactin (PRL) mRNA and its age-related increase (P < 0.0005). Levels of luteinizing hormone (LH) beta and follicle-stimulating hormone (FSH) beta mRNAs did not differ between AL and FR rats until 12 mo, but thereafter rose in FR (LH beta mRNA; P < 0.01, FSH beta mRNA; P < 0.05). Many of these changes in gene expression corroborate previously reported hormonal changes in FR rodents and mutant mice with extended life spans, and thus provide further support for the hypothesis that an altered hormonal milieu contributes to the antiaging effects of food restriction. PMID- 11385055 TI - Short-term orlistat treatment does not affect mineral balance and bone turnover in obese men. AB - Orlistat is a gastrointestinal lipase inhibitor that is used to reduce dietary fat absorption and to enhance weight loss in subjects consuming a hypocaloric diet. To assess whether orlistat has an effect on the metabolism of six minerals, a 21-d, double-blind, randomized, parallel-group, placebo-controlled mineral balance study was conducted in obese (body mass index > 30 kg/m(2)) men. Subjects consumed a hypocaloric diet with a constant daily mineral content and received daily oral treatment with orlistat (120 mg three times daily) (n = 14) or placebo (three times daily) (n = 14) for 21 d. After a 14-d equilibration period, calcium, phosphorus, magnesium, iron, copper and zinc balances were assessed for d 15-21. In addition, the effect of diet and orlistat treatment on bone metabolism was estimated from measurement of biomarkers of bone formation and bone resorption. Serum and urine electrolytes were also measured at baseline and at the end of treatment. Orlistat inhibited fat absorption by approximately 33% (P < 0.05). There were no significant differences in mineral apparent absorption, urinary mineral loss or mineral balance between the orlistat and placebo groups. Markers of bone turnover and serum and urine electrolytes did not differ between the orlistat and placebo groups. Orlistat was well tolerated; adverse events were of mild or moderate intensity, and the majority of these events were unrelated or remotely related to study treatment. In obese men consuming a hypocaloric diet, the administration of orlistat had no significant effect on the balance of six selected minerals. In addition, biomarkers of bone turnover, as well as serum and urine electrolytes, were not affected by orlistat treatment. PMID- 11385056 TI - High linoleic acid, low vegetable, and high oleic acid, high vegetable diets affect platelet activation similarly in healthy women and men. AB - Upregulation of protein kinase C (PKC), an important enzyme in platelet activation, could be one step toward platelet hyperactivity. PKC activation can be modulated by dietary components in vitro, but few data are available concerning the in vivo effects. In this strictly controlled human dietary intervention, the influence of dietary unsaturated fatty acids and vegetable compounds on platelet activation was investigated. A high linoleic acid diet (10% of energy) with small amounts of vegetables (no berries or apples) was consumed by 9 women and 4 men (24.1 +/- 3.9 y), and was compared with a high oleic acid diet (12% of energy) with considerable amounts of vegetables, berries and apples consumed by 8 women and 4 men (24.2 +/- 5.5 y). Subjects were healthy Finnish volunteers. Compliance with the experimental protocol was good, as indicated by changes in plasma fatty acids and concentrations of vitamin C, beta-carotene and alpha-tocopherol. No differences between groups were seen in indices of platelet activation, including platelet aggregation, total PKC activity and distribution of PKC isoenzymes alpha, beta(II) and delta. The results indicate that in apparently healthy and fairly young subjects with adequate vitamin intakes, diets differing markedly in their amounts of linoleic and oleic acids, and vegetables, berries and apples do not differ in platelet activation. PMID- 11385057 TI - The influence of the albumin fraction on the bioavailability and postprandial utilization of pea protein given selectively to humans. AB - Pulse seed proteins such as those found in peas (Pisum sativum) contain fractions of very dissimilar composition and properties, which may therefore be differently utilized by the human body. To analyze the nutritional value of the soluble protein fractions of pea seed, human volunteers ingested a mixed meal of 30 g of raw purified pea protein either as [15N]-globulins (G, n = 9) or as a mix of [15N]-globulins and [15N]-albumins (GA, n = 7) in their natural proportions (22:8). Dietary and endogenous nitrogen fluxes at the terminal ileum were assessed using a tube perfusion technique with an isotopic dilution method. Systemic dietary amino acid availability and the retention of dietary amino acids were determined using 15N enrichment in plasma amino acids and deamination products in blood and urine for 8 h postprandially. The results showed that the pea albumin fraction had the following effects: 1) significantly lowered the real ileal digestibility of pea protein (94 +/- 2.5% for G vs. 89.9 +/- 4% for GA), probably because of a direct effect of trypsin inhibitors; 2) did not promote acute intestinal losses of endogenous nitrogen; and 3) did not significantly improve the postprandial biological value of pea protein (76.5 +/- 3.9% for G vs. 78.7 +/- 3.6% for GA), despite the fact that it corrected the globulin deficiency in sulfur amino acids. We conclude that both G and GA are of good nutritional value for humans and show that cysteine-rich albumins have a far more modest effect on the efficiency of postprandial dietary protein utilization than would be expected from the amino acid scores. PMID- 11385058 TI - A vegetable/fruit concentrate with high antioxidant capacity has no effect on biomarkers of antioxidant status in male smokers. AB - The potential benefits of a high fruit and vegetable intake on the antioxidant status and on relevant biomarkers of oxidative damage to lipids, proteins and DNA and on (functional) markers of oxidative stress were evaluated. A randomized, free living, open placebo-controlled cross-over trial of 3 wk, with a 2-wk washout period between treatments, was performed in a group of 22 male smokers with a relatively low vegetable and fruit intake using a vegetable burger and fruit drink. The vegetable burger and fruit drink increased serum levels of vitamin C, alpha-carotene, beta-carotene, beta-cryptoxanthin and zeaxanthin and plasma total antioxidant capacity. However, no effects were demonstrated on any marker of oxidative damage to lipids (malondialdehyde F(2)-isoprostane) proteins (carbonyls) and DNA (Comet assay) and (functional) markers of oxidative stress (reduced/oxidized glutathione ratio, glutathione-S-transferase alpha, glutathione S-transferase pi and nuclear transcription factor-kappaB). Apparently, these increased levels of antioxidants in serum were not sufficiently high to show beneficial changes with the selected biomarkers. Alternatively, oxidative stress in male smokers with a relatively low fruit and vegetable intake might have been still too low to demonstrate a beneficial effect of antioxidants. PMID- 11385059 TI - Long-term pharmacologic doses of vitamin E only moderately affect the erythrocytes of patients with type 1 diabetes mellitus. AB - In erythrocytes from diabetic patients, increased membrane lipid peroxidation might lead to abnormalities in composition and function. To study this relationship, we investigated the effects of a moderate pharmacologic dose of vitamin E for 1 y on erythrocyte membrane peroxidation in vitro and on its fatty acid composition, antioxidant capacity and rheological function. In a random and double-blind manner, type 1 diabetic patients (n = 44) were assigned to the following two groups: Group S received 250 IU (168 mg) d-alpha tocopherol 3 times daily for 1 y. Group P received placebo for 6 mo followed by d-alpha-tocopherol for an additional 6 mo. Variables were monitored every 3 mo. After 3 mo of supplementation, serum vitamin E doubled (P < 0.0005), thiobarbituric acid reactive substances in erythrocyte membranes incubated with tert-butyl hydroperoxide decreased by 25% (P = 0.006) and the lagtime of fluorescence increased from 28 +/- 16 to 41 +/- 28 min (P = 0.028). Patients who did not respond to supplementation (13 of 44) had lower serum lipids (P = 0.017) and body mass index (P = 0.024). We did not detect any significant effects of vitamin E supplementation on membrane lipid composition, antioxidant capacity or blood viscosity. Continuing supplementation for up to 1 y did not further affect serum vitamin E or membrane peroxidation. Stopping supplementation was followed by a return to inclusion values. These results show that the decrease in erythrocyte membrane peroxidation after vitamin E supplementation is moderate, saturable, reversible, restricted to some individuals and has no detectable effect on erythrocyte composition and function. PMID- 11385060 TI - Catechins are bioavailable in men and women drinking black tea throughout the day. AB - Tea consumption has been associated with reduced risk of both cancer and cardiovascular disease in population studies, but clinical data demonstrating bioavailability of the individual catechins and other polyphenolic components of tea are limited. This study assessed the apparent bioavailability of the prominent catechins from black tea in humans drinking tea throughout the day. After 5 d of consuming a low flavonoid diet, subjects drank a black tea preparation containing 15.48, 36.54, 16.74, and 31.14 mg of (-)-epigallocatechin (EGC), (-)-epicatechin (EC), (-)-epigallocatechin gallate (EGCG) and (-) epicatechin gallate (ECG), respectively, at four time points (0, 2, 4 and 6 h). Blood, urine and fecal specimens were collected over a 24- to 72-h period and catechins were quantified by HPLC with coularray detection. Plasma concentrations of EGC, EC and EGCG increased significantly relative to baseline (P < 0.05). Plasma EGC, EC and EGCG peaked after 5 h, whereas ECG peaked at 24 h. Urinary excretion of EGC and EC, which peaked at 5 h, was increased relative to baseline amounts (P < 0.05) and fecal excretion of all four catechins was increased relative to baseline (P < 0.05). Approximately 1.68% of ingested catechins were present in the plasma, urine and feces, and the apparent bioavailability of the gallated catechins was lower than the nongallated forms. Thus, catechins were bioavailable. However, unless they are rapidly metabolized or sequestered, the catechins appeared to be absorbed in amounts that were small relative to intake. PMID- 11385061 TI - Food insecurity is positively related to overweight in women. AB - Although individuals with poor food security might be expected to have reduced food intake, and thus reduced body fat and less likelihood of being overweight, these associations have not been adequately studied. The purpose of the current study was to examine the relationship between food insecurity and overweight as measured by body mass index (BMI) using data from the nationally representative 1994-1996 Continuing Survey of Food Intakes by Individuals (CSFII). Overweight was defined as BMI >27.3 kg/m(2) for women and 27.8 kg/m(2) for men. Food insecurity was related to overweight status for women (n = 4509, P < 0.0001), but not for men (n = 4970, P = 0.44). Excluding the 11 severely insecure women, the prevalence of overweight among women increased as food insecurity increased, from 34% for those who were food secure (n = 3447), to 41% for those who were mildly food insecure (n = 966) and to 52% for those who were moderately food insecure (n = 86). Food insecurity remained a significant predictor of overweight status, after adjustment for potentially confounding demographic and lifestyle variables (P < 0.01). In a logistic regression analysis, mildly insecure women were 30% more likely to be overweight than those who were food secure [odds ratio (OR) 1.3, P = 0.005]. Thus, food insecurity had an unexpected and paradoxical association with overweight status among women with a higher prevalence of overweight among the food insecure, and a resulting potential for increased incidence of obesity-related chronic diseases. Given that the rates of both overweight and food insecurity are on the rise, this research area warrants further investigation. PMID- 11385062 TI - Testosterone and food restriction modulate hepatic lycopene isomer concentrations in male F344 rats. AB - We previously demonstrated that the castration of male rats profoundly increases hepatic lycopene compared with intact controls. Here we further characterized the role of testosterone in modulating hepatic lycopene accumulation and isomer patterns in male rats. Furthermore, because castration significantly decreases ad libitum food consumption, we investigated the influence of food restriction on lycopene metabolism. Forty male F344 rats 8 wk of age were randomly assigned to one of four treatments (n = 10/group): 1) intact, free access to food, 2) castration, free access to food, 3) castration plus testosterone implants, free access to food and 4) intact, 20% food restricted. All rats were fed an AIN-based diet with 0.25 g lycopene (as 10% water-soluble beadlets)/kg diet for 3 wk. Serum testosterone was 5.31 +/- 1.46 nmol/L in intact controls allowed free access to food, reduced in castrated animals (0.52 +/- 0.10, P < 0.0001 versus controls) and intact, food-restricted rats (1.53 +/- 0.49 nmol/L, P < 0.0001 versus controls) and greater (17.23 +/- 3.09 nmol/L) in castrated rats administered testosterone (P < 0.0001 versus controls). Castrated rats accumulated approximately twice as much liver lycopene (74.5 +/- 8.5 nmol/g; P < 0.01 versus controls) as intact rats allowed free access to food (39.5 +/- 5.0) despite 13% lower dietary lycopene intake (P < 0.001; 3.38 +/- 0.07 versus 3.95 +/- 0.06 mg lycopene/d). Testosterone replacement in castrated rats returned liver lycopene concentrations (32.5 +/- 5.5 nmol lycopene/g with 3.76 +/- 0.05 mg dietary lycopene/d) to those observed in intact rats. Food restriction resulted in a 20% decrease in lycopene intake but significantly increased liver lycopene by 68% (66.3 +/- 7.9 nmol lycopene/g with 3.38 +/- 0.00 mg lycopene/d) compared with controls and castrated rats administered testosterone. These results suggest that androgen depletion and 20% food restriction increase hepatic lycopene accumulation. We hypothesize an endocrine and dietary interaction, where higher androgen concentrations and greater energy intake may stimulate lycopene metabolism and degradation. PMID- 11385063 TI - Catechin is metabolized by both the small intestine and liver of rats. AB - Flavan-3-ols are the most abundant flavonoids in the human diet, but little is known about their absorption and metabolism. In this study, the absorption and metabolism of the monomeric flavan-3-ol, catechin, was investigated after the in situ perfusion of the jejunum + ileum in rats. Five concentrations of catechin were studied, ranging from 1 to 100 micromol/L. The absorption of catechin was directly proportional to the concentration, and 35 +/- 2% of the perfused catechin was absorbed during the 30-min period. Effluent samples contained only native catechin, indicating that intestinal excretion of metabolites is not a mechanism of catechin elimination. Catechin was absorbed into intestinal cells and metabolized extensively because no native catechin could be detected in plasma from the mesenteric vein. Mesenteric plasma contained glucuronide conjugates of catechin and 3'-O-methyl catechin (3'OMC), indicating the intestinal origin of these conjugates. Additional methylation and sulfation occurred in the liver, and glucuronide + sulfate conjugates of 3'OMC were excreted extensively in bile. Circulating forms were mainly glucuronide conjugates of catechin and 3'OMC. The data further demonstrate the role of the rat small intestine in the glucuronidation and methylation of flavonoids as well as the role of the liver in sulfation, methylation and biliary excretion. PMID- 11385064 TI - Low fat and high monounsaturated fat diets decrease human low density lipoprotein oxidative susceptibility in vitro. AB - Oxidative modification of low density lipoprotein (LDL) is thought to play an important role in the development of atherosclerosis. Some studies have found that LDL enriched in monounsaturated fatty acids (MUFA) are less susceptible to oxidation than LDL enriched in polyunsaturated fatty acids (PUFA). A high MUFA diet is an alternative to a lower-fat blood cholesterol-lowering diet. Less is known about the effects of high MUFA versus lower-fat blood cholesterol-lowering diets on LDL oxidative susceptibility. The present study was designed to evaluate the effects of men and women consuming diets high in MUFA (peanuts plus peanut butter, peanut oil and olive oil) on LDL oxidative susceptibility, and to compare these effects with those of a Step II blood cholesterol-lowering diet. A randomized, double-blind, five-period crossover design (n = 20) was used to study the effects of the following diets on LDL-oxidation: average American [35% fat, 15% saturated fatty acids (SFA)], Step II (25% fat, 7% SFA), olive oil (35% fat, 7% SFA), peanut oil (35% fat, 7% SFA) and peanuts plus peanut butter (35% fat, 8% SFA). The average American diet resulted in the shortest lag time (57 +/- 6 min) for LDL oxidized ex vivo, whereas the Step II, olive oil and peanuts plus peanut butter diets resulted in a lag time of 66 +/- 6 min (P < or = 0.1). The slower rate of oxidation [nmol dienes/(min x mg LDL protein)] observed when subjects consumed the olive oil diet (24 +/- 2) versus the average American (28 +/- 2), peanut oil (28 +/- 2) and peanuts plus peanut butter diets (29 +/- 2; P < or = 0.05) was associated with a lower LDL PUFA content. The results of this study suggest that lower-fat and higher-fat blood cholesterol-lowering diets high in MUFA have similar effects on LDL oxidative resistance. In addition, our results suggest that different high MUFA sources varying in the ratio of MUFA to PUFA can be incorporated into a high MUFA diet without increasing the susceptibility of LDL to oxidation. PMID- 11385065 TI - Milk fat synthesis in dairy cows is progressively reduced by increasing supplemental amounts of trans-10, cis-12 conjugated linoleic acid (CLA). AB - Conjugated linoleic acid (CLA) supplements containing a variety of isomers reduce milk fat yield. We have recently identified trans-10, cis-12 CLA as the isomer responsible for inhibiting milk fat synthesis in dairy cows. Our objectives were to determine milk fat yield and fatty acid composition responses to different doses of trans-10, cis-12 CLA. Multiparous Holstein cows (n = 4) were used in a 4 x 4 Latin square design. Treatments consisted of a 5-d abomasal infusion of four doses of trans-10, cis-12 CLA, i.e., 0.0, 3.5, 7.0 and 14.0 g/d. Milk fat yield was decreased 25, 33, and 50%, and milk fat concentration was reduced 24, 37 and 46% when cows received 3.5, 7.0 and 14.0 g/d of trans-10, cis-12 CLA, respectively. Feed intake, milk yield, and milk protein content and yield were unaffected by treatment. Milk fatty acid composition revealed that de novo synthesized fatty acids (short and medium chain) were extensively reduced when cows received the two highest doses, but at the low dose (3.5 g/d), decreases in de novo synthesized fatty acids and preformed fatty acids were similar. Changes in milk fatty acid composition also demonstrated that (9)-desaturase activity was inhibited at the two high doses of trans-10, cis-12 CLA, but was unaffected by the low dose. Results indicate minimal quantities of trans-10, cis-12 CLA (0.016% of dietary dry matter) markedly inhibited milk fat synthesis (25% reduction) and that a curvilinear reduction in milk fat yield occurred with increasing quantities of trans-10, cis-12 CLA. PMID- 11385066 TI - Whole wheat and triticale flours with differing viscosities stimulate cecal fermentations and lower plasma and hepatic lipids in rats. AB - Whole flours from oat, rye or barley effectively modify digestive fermentation and lipid metabolism, whereas the effectiveness of whole wheat flour has not been established. To address this question, cecal digestion, short-chain fatty acid (SCFA) metabolism and cholesterol metabolism were investigated in four groups of rats fed the following semipurified diets differing in their carbohydrate source: a control diet (purified wheat starch) and three whole cereal flour diets [Valoris wheat (Wv), Soissons wheat (Ws), or Carnac triticale (Tc)]. Wv is particularly viscous and rich in arabinoxylans, and Tc is richer in hemicellulose than wheat. Compared with controls, rats fed the whole-flour diets had enlarged ceca and a moderate acidification of the bulk pH ( approximately 6.4). In these rats, the cecal SCFA pool size was enhanced (P < 0.05), and the SCFA molar ratio reflected propionic/butyric acid-rich fermentations, especially in those fed TC: The portal SCFA concentrations reflected the rise of the acetic and propionic acid pools in the cecum, whereas portal butyric acid remained relatively low, probably reflecting extensive metabolism by the cecal wall. The fecal excretion of total steroids (bile acids + sterols) was markedly enhanced by all of the whole-flour diets, with Wv (+78%) > Tc (+64%) > Ws (+47%). In parallel, there was a significant plasma cholesterol-lowering effect for rats fed Wv (-27%) and Tc ( 32%) and a plasma triglyceride-lowering effect (approximately -40%) in all rats fed whole-flour diets (P < 0.05). This effect was observed mainly for triglyceride-rich lipoprotein-cholesterol, whereas HDL cholesterol was unaffected. These results indicate that whole wheat flours can strikingly affect cecal SCFA, especially butyrate, and are effective plasma cholesterol-lowering agents. PMID- 11385067 TI - Assessment of vitamin B-6 status in young women consuming a controlled diet containing four levels of vitamin B-6 provides an estimated average requirement and recommended dietary allowance. AB - The Recommended Dietary Allowance (RDA) of vitamin B-6 for young women was recently reduced from 1.6 to 1.3 mg/d based on an adequate plasma pyridoxal phosphate (PLP) concentration of 20 nmol/L. To assess vitamin B-6 requirements and suggest recommendations for intake, seven healthy young women consumed a controlled diet providing 1.2 g protein/kg body weight for a 7-d adjustment period (1.0 mg vitamin B-6/d) and three successive 14-d experimental periods (1.5, 2.1 and 2.7 mg/d, respectively). Direct and indirect vitamin B-6 status indicators were measured in plasma, erythrocytes and urine. Indicators most strongly correlated with vitamin B-6 intake [i.e., plasma and erythrocyte PLP, urinary 4-pyridoxic acid (4-PA) and total vitamin B-6] were regressed on vitamin B-6 intake and the dietary vitamin B-6 to protein ratio. Inverse prediction using adequate and baseline values estimated vitamin B-6 requirement. Adequate values were determined for plasma PLP and urinary 4-PA from baseline values of 60 previous subjects, using the statistical method suggested by Sauberlich. The current study suggests a vitamin B-6 Estimated Average Requirement (EAR) for young women of 1.1 mg/d or 0.016 mg/g protein, and a RDA of 1.5 mg/d or 0.020 mg/g protein. When results from this study are combined with data from four other recent studies, the combined data predict an EAR of 1.2 mg/d or 0.015 mg/g protein, and a RDA of 1.7 mg/d or 0.018 mg/g protein. This study suggests that the current vitamin B-6 RDA may not be adequate. PMID- 11385068 TI - Vitamin D receptor null mutant mice fed high levels of calcium are fertile. AB - Vitamin D receptor (VDR) null mutant mice provide a model to investigate the possible effect of vitamin D on female reproduction. Infertility in these mice has been reported but it is uncertain whether the infertility results from a lack of VDR or from the hypocalcemia that results from a lack of VDR. VDR null mutant mice and wild-type controls were fed a nonpurified, high calcium or medium calcium diet, plus a diet containing lactose and their reproductive efficiency was examined. VDR null mutant mice fed a nonpurified diet were hypocalcemic and were found to be largely infertile with 14% fertility, while the fertility percentage of normocalcemic VDR null mutant mice and wild-type mice was between 86% and 100%. A high calcium or medium calcium diet maintained 100% fertility in the VDR knockout mice; removal of the lactose from this diet did not diminish reproductive capability. Reproductive capacity of VDR null mutant mice was analyzed when they were fed purified diets containing 0.02-2% calcium. Mutant mice fed a low calcium diet (0.47%) had a lower reproductive efficiency than VDR null mutant mice fed a diet that resulted in normal serum calcium concentrations. Thus, high dietary calcium levels are required for normal reproduction in VDR null mutant female mice. It seems that the defect in reproduction reported previously for VDR null mutant mice is not the lack of a direct effect of 1,25 dihydroxycholecalciferol on reproductive function but is the result of hypocalcemia. PMID- 11385069 TI - Microbial phytase does not improve protein-amino acid utilization in soybean meal fed to young chickens. AB - Three growth trials were conducted with young chicks to evaluate crude protein (CP) utilization in soybean meal (SBM) as affected by dietary addition of microbial phytase. In assay 1, chicks were fed two CP-deficient (50 or 150 g CP/kg) levels of dehulled SBM, and each SBM level was then supplemented with equimolar amounts of cystine or methionine (Met) or with 1200 U phytase/kg. At 50 g CP/kg, cystine or Met supplementation improved (P < 0.05) measures of growth performance, but when 150 g CP/kg from SBM was fed, only Met addition improved (P < 0.05) weight gain, food efficiency and protein efficiency ratio (PER). Thus, Cys was more limiting than Met in the diet that contained 50 g CP/kg, but Met was clearly first-limiting in the diet that contained 150 g CP/kg. Phytase supplementation did not improve (P > 0.10) chick performance at either level of CP. Chicks in assay 2 were fed 100 g CP/kg furnished by SBM, casein or corn gluten meal in the absence and presence of 1200 U phytase/kg. Weight gain, gain/food and PER values were greater (P < 0.05) in chicks fed SBM than in those fed casein, and greater (P < 0.05) in chicks fed casein than in those fed corn gluten meal. Phytase supplementation had no effect (P > 0.10) on any measure of chick performance, regardless of the protein source fed. In assay 3, three deficient levels of CP (50, 100 and 150 g/kg) from SBM were fed in the absence and presence of 1200 U dietary phytase/kg. Weight gain, food efficiency and protein accretion increased linearly (P < 0.05) as a function of protein intake, but phytase supplementation had no effect (P > 0.10) on slopes of the body weight and protein accretion curves. Likewise, phytase addition did not affect (P > 0.10) measures of protein utilization, i.e., weight gain/protein intake and protein gain/protein intake at any of the CP levels that were fed. Because sulfur amino acids are the growth-limiting factors when protein-deficient levels of SBM are fed to young chicks, we conclude that dietary addition of phytase does not improve sulfur amino acid utilization in SBM. PMID- 11385070 TI - Combined selenium and vitamin E deficiency causes fatal myopathy in guinea pigs. AB - Selenium and vitamin E deficiencies were studied as part of an evaluation of oxidant defenses in guinea pigs. Male guinea pigs (100-120 g) were fed a control diet (C) or the diet without selenium (0 Se), without vitamin E (0 E), or without either selenium or vitamin E (0 Se-0 E). Between d 30 and 35, 7 of 13 guinea pigs fed the 0 Se-0 E diet were euthanized because of severe weakness of their extremities. No guinea pigs in the other diet groups developed weakness. Guinea pigs from each group were killed on d 37. Selenium deficiency and vitamin E deficiency were verified by measurement of glutathione peroxidase and alpha tocopherol. Creatine phophokinase (CPK) activity was greater than controls in both groups fed vitamin E-deficient diets, but the increase was greater in the 0 Se-0 E group than in the 0 E group. Muscle F(2)-isoprostanes were greater than controls in both groups fed vitamin E-deficient diets with the level in the 0 Se 0 E group greater than that in the 0 E group. Histologic muscle necrosis was severe in the 0 Se-0 E group, minimal in the 0 E group and absent from other groups. The diets used in this study induced selenium and vitamin E deficiencies in guinea pigs. The study demonstrates that combined selenium and vitamin E deficiency results in a fatal myopathy in guinea pigs that is associated with lipid peroxidation in the affected muscle. This nutritional myopathy is much more severe than the myopathy that occurs with vitamin E deficiency alone. PMID- 11385071 TI - Hamsters predisposed to sucrose-induced cholesterol gallstones (LPN strain) are more resistant to excess dietary cholesterol than hamsters that are not sensitive to cholelithiasis induction. AB - We compared the effects of cholesterol feeding in male hamsters from two strains with different propensities to sucrose-induced cholelithiasis; Laboratoire de Physiologie de la Nutrition (LPN) hamsters are predisposed to developing biliary cholesterol gallstones, whereas Janvier (JAN) hamsters are not. When fed a basal control diet, LPN hamsters had a lower cholesterolemia (-21%, P = 0.01) than JAN hamsters, and a higher activity of 3-hydroxy-3-methyl glutaryl coenzyme A reductase in liver (+148%, P = 0.018) and intestine (+281%, P < 0.0001). After feeding the same diet enriched with 0.3% cholesterol for 5 wk, cholesterolemia increased more dramatically in JAN hamsters (+235%, P < 0.001) than in LPN hamsters (+108%, P < 0.001), as did the liver concentration of cholesterol, which reached 152.30 +/- 13.00 and 44.41 +/- 9.06 micromol/g, respectively. Only JAN hamsters displayed hepatomegaly, with an increased cholesterol saturation index of the gallbladder bile (+100%, P < 0.01), due to the cholesterol challenge. In liver, cholesterol feeding reduced cholesterol 7alpha-hydroxylase activity and mRNA level, and stimulated sterol 27-hydroxylase and oxysterol 7alpha-hydroxylase activities. Hepatic levels of LDL receptor decreased by approximately 60% in both strains, whereas HDL receptor scavenger class B type 1 (SR-BI) levels were unaffected by dietary cholesterol. The greater resistance of LPN hamsters to the hypercholesterolemic diet can be explained by a lower capacity to store cholesterol in the liver and greater efficiency in reducing the activity of 3 hydroxy-3-methyl glutaryl coenzyme A reductase in response to cholesterol feeding [from 11263 to 261 pmol/(min x organ) in LPN hamsters and from 4530 to 694 pmol/(min x organ) in JAN hamsters]. These results highlight the usefulness of this two-strain model, which offers some analogy with the inverse association between the predisposition to cholelithiasis and the risk of atherosclerosis in humans. PMID- 11385072 TI - Overabundance of CD45RA(+) (quiescent-phenotype) cells within the involuted CD4(+) T-cell population follows initiation of immune depression in energy deficient weanling mice and reflects involution exclusive to the CD45RA(-) subset. AB - Previous studies have identified an overabundance of quiescent-phenotype (CD45RA(+)) CD4(+) T cells throughout the lymphoid system of weanling mice at an advanced stage of food intake restriction mimicking marasmus. The objective of this investigation was to determine the timing of this phenomenon relative to the development of depression in cell-mediated immune competence. Two experiments were conducted in which male and female weanling C57BL/6J mice, initially 19 d of age, either were permitted free access to a complete purified diet or were subjected to restricted intake of this diet, producing loss of 1.5-2% of initial body weight daily. In the first experiment, feeding periods of 3, 6, 9, 12 and 14 d were examined, and a zero-time control group (19 d old) was also included. Expression of CD45RA was assessed by flow cytometry in CD4(+) T cells from the blood, spleen and mesenteric lymph nodes. Despite reduction in CD4(+) T-cell numbers, evident in all three lymphoid compartments of the malnourished mice by d 6, energy-restricted mice maintained the numbers of CD4(+)CD45RA(+) T cells at the level found in the zero-time control group. Consequently, the malnourished group exhibited a high percentage of CD4(+) T cells expressing CD45RA by d 9 in the blood and mesenteric nodes and by d 12 in the spleen. In the second study, malnourished and age-matched control groups were sensitized to sheep red blood cells on d 3 and energy-restricted mice exhibited depression in the delayed hypersensitivity response to this antigen when assessed on d 9 after challenge 24 h previously. Energy deficiency pathology includes a shift toward CD4(+) T cell quiescence that may contribute to ongoing immunodepression without being involved in its initiation. Remarkably, this imbalance develops because involution of the CD4(+) subset in the energy-deficient mice is confined to the CD45RA(-) population. PMID- 11385073 TI - An interaction between hypothalamic glucagon-like peptide-1 and macronutrient composition determines food intake in rats. AB - Glucagon-like peptide-1 (GLP-1) release in response to food ingestion has been associated with decreased food intake. In the present study, we tested the hypothesis that the feeding response to GLP-1 injection into the hypothalamic paraventricular nucleus (PVN) is influenced by the macronutrient composition of the food consumed. In the first experiment, rats were injected with GLP-1 (0.2 microg) or saline (0.5 microL) in the PVN at dark onset (1800 h), and food intake from a maintenance diet (18% protein) was measured at 1, 2 and 14 h. In Experiment 2, after GLP-1 injection, rats were fed a carbohydrate (protein-free) diet for the first 2 h or gavaged with glucose (1.4 g/5 mL). In Experiment 3, after GLP-1 injection, rats were fed a protein (50%) diet for the first 2 h, or were preloaded with egg albumin (1.0 g). In the last experiment, GLP-1 was given after corn oil gavage (2.4 g). GLP-1 injection resulted in a reduced consumption of the maintenance diet from 2 to 14 h. The decreased food intake from 2 to 14 h after GLP-1 administration occurred after carbohydrate intake, either by meal or preloads, but not after protein intake, either as a meal or preload. A transient interaction of GLP-1 with a corn oil gavage was detected but only in early feeding (0-2 h). We conclude that the effect of GLP-1 injected in the PVN on food intake is influenced by the macronutrient composition of the food consumed. Carbohydrate enhances, protein blocks and corn oil has a transient effect on the suppression of food intake caused by GLP-1 in the PVN. PMID- 11385074 TI - Intake of dietary phytoestrogens is low in postmenopausal women in the United States: the Framingham study(1-4). AB - Many plants that are consumed contain phytoestrogens. Only a few published studies have examined the dietary intake of phytoestrogens in the general Western population. The potentially positive health effects of phytoestrogens might be of relevance to postmenopausal women. The aim of the present study was to estimate the intake of dietary isoflavones, coumestans and lignans by healthy Western postmenopausal women. For this purpose, we studied 964 postmenopausal, Caucasian women who participated in the Framingham Offspring Study and completed the Willett food-frequency questionnaire (FFQ). By searching the medical and agricultural literature and contacting experts, we identified food sources of phytoestrogens. The concentrations of the different isoflavones, coumestrol and lignans in each food in the FFQ were scored in seven categories and multiplied by the serving size of the food and the frequency of its consumption. The estimated daily median intake of the isoflavone daidzein was 39 microg (24-57 microg); of genistein, 70 microg (28-120 microg); of formononetin, 31 microg (13-44 microg); and of biochanin A, 6 microg (2-11 microg). Median total intake of isoflavones was 154 microg (99-235 microg). The main sources of isoflavones were beans and peas. The estimated daily intake of coumestans was 0.6 microg (0.2-1.7 microg), with broccoli as the main source. The estimated daily median intake of matairesinol was 19 microg (12-28 microg) and of secoisolariciresinol 560 microg (399-778 microg). The median total intake of lignans was 578 microg (416-796 microg). The main source of the lignans was fruits. The daily dietary intake of phytoestrogens in healthy postmenopausal Caucasian women in the United States is <1 mg. PMID- 11385075 TI - An underfeeding study in healthy men and women provides further evidence of impaired regulation of energy expenditure in old age. AB - The effect of aging on energy regulation remains controversial. We compared the effects of underfeeding on changes in energy expenditure and respiratory quotient in young normal weight men and women [YNW, age 25.7 +/- 3.2 y(SD), body mass index (BMI) 23.1 +/- 1.6 kg/m(2)], young overweight men and women (YOW, age 26.1 +/- 3.5 y, BMI 27.7 +/- 2.1 kg/m(2)) and older (OLD) men and women (age 68.4 +/- 3.3 y, BMI 27.4 +/- 3.4 kg/m(2)). The thermic effect of feeding (TEF) during weight maintenance, and changes in resting energy expenditure (REE) and respiratory quotient were determined in response to undereating by an average 3.75 MJ/d for 6 wk. In addition, body composition was measured. No significant differences among the groups were observed in TEF, fasting and postprandial respiratory quotient, or the change in fasting respiratory quotient with underfeeding. However, REE adjusted for fat-free mass and fat mass was significantly lower in OLD subjects compared with YNW and YOW subjects (P < 0.05). In addition, the REE response to weight change was significantly attenuated in the OLD subjects (P = 0.023). These data suggest that the responsiveness of energy expenditure to negative energy balance is attenuated in old age, and provide further support for the hypothesis that mechanisms of energy regulation are broadly disregulated in old age. PMID- 11385076 TI - Tributyrin, a stable and rapidly absorbed prodrug of butyric acid, enhances antiproliferative effects of dihydroxycholecalciferol in human colon cancer cells. AB - Tributyrin, a prodrug of natural butyrate, has been evaluated with an aim to overcome pharmacokinetic drawbacks of natural butyrate as a drug, i.e., its rapid metabolization and inability to achieve pharmacologic concentrations in neoplastic cells. We studied the effects of tributyrin on growth, differentiation and vitamin D receptor expression in Caco-2 cells, a human colon cancer cell line. Tributyrin was more potent in inhibiting growth and inducing cell differentiation than natural butyrate. The effect was further enhanced after addition of physiologic concentrations of dihydroxycholecalciferol [(OH)2D3]. The synergistic effect of tributyrin and (OH)2D3 in Caco-2 cells was due to tributyrin-induced overexpression of the vitamin D receptor, as measured by reverse transcriptase-polymerase chain reaction. Treatment with tributyrin increased binding of (OH)2D3 to its receptor 1.5-fold, without any change in receptor affinity. We conclude that tributyrin may, at least in part, exert its growth-reducing and differentiation-inducing effect in Caco-2 cells by an upregulation of the vitamin D receptor; this may provide a useful therapeutic approach in chemoprevention and treatment of colorectal cancer by the two nutrients occurring naturally in human diet. PMID- 11385077 TI - Resveratrol isolated from Polygonum cuspidatum root prevents tumor growth and metastasis to lung and tumor-induced neovascularization in Lewis lung carcinoma bearing mice. AB - Resveratrol is a naturally occurring phytoalexine found in medicinal plants. We found that resveratrol, at doses of 2.5 and 10 mg/kg, significantly reduced the tumor volume (42%), tumor weight (44%) and metastasis to the lung (56%) in mice bearing highly metastatic Lewis lung carcinoma (LLC) tumors, but not at a dose of 0.6 mg/kg. Resveratrol did not affect the number of CD4(+), CD8(+) and natural killer (NK)1.1.(+) T cells in the spleen. Therefore, the inhibitory effects of resveratrol on tumor growth and lung metastasis could not be explained by natural killer or cytotoxic T-lymphocyte activation. In addition, resveratrol inhibited DNA synthesis most strongly in LLC cells; its 50% inhibitory concentration (IC(50)) was 6.8 micromol/L. Resveratrol at 100 micromol/L increased apoptosis to 20.6 +/- 1.35% from 12.1 +/- 0.36% (P < 0.05) in LLC cells, and decreased the S phase population to 22.1 +/- 1.03% and 29.2 +/- 0.27% from 35.2 +/- 1.72% (P < 0.05) at concentrations of 50 and 100 micromol/L, respectively. Resveratrol inhibited tumor-induced neovascularization at doses of 2.5 and 10 mg/kg in an in vivo model. Moreover, resveratrol significantly inhibited the formation of capillary-like tube formation from human umbilical vein endothelial cells (HUVEC) at concentrations of 10-100 micromol/L; the degree of the inhibition of capillary like tube formation by resveratrol was 45.5% at 10 micromol/L, 50.2% at 50 micromol/L and 52.6% at 100 micromol/L. Resveratrol inhibited the binding of vascular endothelial growth factor (VEGF) to HUVEC at concentrations of 10-100 micromol/L, but not at concentrations of 1 and 5 micromol/L. The degree of inhibition of VEGF binding to HUVEC by resveratrol was 16.9% at 10 micromol/L, 53.2% at 50 micromol/L and 47.8% at 100 micromol/L. We suggest that the antitumor and antimetastatic activities of resveratrol might be due to the inhibition of DNA synthesis in LLC cells and the inhibition of LLC-induced neovascularization and tube formation (angiogensis) of HUVEC by resveratrol PMID- 11385078 TI - A buckwheat protein product suppresses 1,2-dimethylhydrazine-induced colon carcinogenesis in rats by reducing cell proliferation. AB - This study was conducted to examine the effect of consumption of buckwheat protein product (BWP) on 1,2-dimethylhydrazine (DMH)-induced colon tumor in rats. Male growing Sprague-Dawley rats were fed diets containing either casein or BWP (net protein level, 200 g/kg; n = 20/group) for 124 d. The rats were gavaged weekly with DMH (20 mg/kg body) for the first 8 wk. Food intake and growth were unaffected by dietary manipulation. Dietary BWP caused a 47% reduction in the incidence of colonic adenocarcinoma (P < 0.05), but did not affect the incidence of colonic adenomas. BWP intake tended to reduce the number of colon adenocarcinomas (P = 0.16). Consumption of BWP significantly reduced cell proliferation and expression of c-myc and c-fos proteins in colonic epithelium. The results suggest that dietary BWP has a protective effect against DMH-induced colon carcinogenesis in rats by reducing cell proliferation. PMID- 11385079 TI - Accuracy of simple techniques for estimating fractional zinc absorption in humans. AB - The theoretical basis of the accuracy of a number of simple techniques for estimating fractional zinc absorption (FZA) in humans using stable isotopic tracers has not been evaluated. These techniques include fecal monitoring (FM), deconvolution analysis (DA), double isotopic tracer ratio (DITR) and indicator dilution methods. Using a compartmental model, we investigated the accuracy and logic of each of these techniques. Time-dependent estimates of FZA based on the simple techniques were simulated using the compartmental model and compared with the known FZA derived from the model. The analysis elucidated logical errors in some of the FM techniques, and even when these problems were corrected, the FM technique was still prone to errors due to incomplete fecal tracer recovery and variable gastrointestinal (GI) transit time. Although logically correct, the indicator dilution techniques were also highly sensitive to incomplete fecal tracer recovery and variable GI transit time. The DA and DITR techniques were the most robust in that they were logically correct and were insensitive to incomplete fecal tracer recovery and variable GI transit time. Although all of the DA and DITR methods provided similarly good estimates of FZA relative to the compartmental model, the DITR technique performed on a spot urine specimen obtained several days after tracer administration was the preferred choice because of its simplicity and minimal requirements for patient compliance. PMID- 11385080 TI - Molecular ecological analysis of dietary and antibiotic-induced alterations of the mouse intestinal microbiota. AB - A cultivation-independent approach, polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE), was used to characterize changes in fecal bacterial populations resulting from consumption of a low residue diet or oral administration of a broad-spectrum antibiotic. C57BL/6NHsd mice were weaned to either a standard nonpurified diet (LC-diet) or a low residue diet (LR-diet) and at 17 wk of age were randomly assigned to receive drinking water with or without 25 ppm cefoxitin for 14 d. On d 1, 2, 7 and 14, microbial DNA was extracted from feces, and the V3 region of the 16S rDNA gene was amplified by PCR and analyzed by DGGE. The diversity of fecal microbial populations, assessed using Shannon's index (H'), which incorporates species richness (number of species, or in this case, PCR-DGGE bands) and evenness (the relative distribution of species), was not affected by cefoxitin. However, use of Sorenson's pairwise similarity coefficient (C(s)), an index that measures the species in common between different habitats, indicated that the species composition of fecal bacterial communities was altered by cefoxitin in mice fed either diet. Dietary effects on fecal microbial communities were more pronounced, with greater H' values (P < 0.05) in mice fed the LR-diet (1.9 +/- 0.1) compared with the LC-diet (1.6 +/- 0.1). The C(s) values were also greater (P < 0.05) in fecal bacterial populations from mice fed the LR-diet (C(s) = 69.8 +/- 2.0%) compared with mice fed the LC diet (C(s) = 50.1 +/- 3.8%), indicating greater homogeneity of fecal bacterial communities in mice fed the LR-diet. These results demonstrate the utility of cultivation-independent PCR-DGGE analysis combined with measurements of ecological diversity for monitoring diet- and antibiotic-induced alterations of the complex intestinal microbial ecosystem. PMID- 11385081 TI - Dietary selenium and arsenic affect DNA methylation. PMID- 11385085 TI - A new dimension for the JRSM. PMID- 11385084 TI - BSE and vCJD: what is the future? PMID- 11385086 TI - Management of diabetic nephropathy. PMID- 11385087 TI - Surgery for intersex. PMID- 11385088 TI - The renal tubular acidoses. PMID- 11385089 TI - Reducing risk by improving standards of intrapartum fetal care. AB - Confidential Enquiries into Stillbirths and Deaths in Infancy (CESDI) have pointed to a high frequency of suboptimal intrapartum fetal care of a kind that, in the event of an adverse outcome, is hard to defend in court. In an effort to minimize liability, various strategies were applied in a district hospital labour ward--guidelines, cyclical audit, monthly feedback meetings and training sessions in cardiotocography (CTG). The effects of these interventions on quality of care was assessed by use of the CESDI system in all babies born with an Apgar score of 4 or less at 1 min and/or 7 or less at 5 min. 540 babies (4.3%) had low Apgar scores, and neither the percentage nor gestational age differed significantly between audit periods. In the baseline audit, care was judged suboptimal (grade II/III) in 14 (74%) of 19 cases, and in the next four periods it was 23%, 27%, 27% and 32%. In the latest audit period, after further educational interventions, it was 9%. Many of the failures to recognize or act on abnormal events were related to CTG interpretation. After the interventions there was a significant increase in cord blood pH measurement. There were no differences between audit periods in the proportion of babies with cord pH < 7.2. These results indicate that substantial improvements in quality of intrapartum care can be achieved by a programme of clinical risk management. PMID- 11385090 TI - Surgical decisions in the elderly: the importance of biological age. AB - Surgeons will sometimes advise against an operation because the patient is 'old and frail'. A simple starring system (one to five), based on performance and lifestyle, has been devised to assess the biological age of elderly patients. 10 consultant surgeons and 10 trainees answered questions about their treatment recommendations for hypothetical patients of standard age and medical history but with various star ratings and surgical conditions. 1000 decisions were available for analysis. The four and five star patients (those leading an independent existence) were recommended 266 interventions, the one and two star patients 55. Trainees were more inclined to intervene than consultants, recommending operations in half the patients rather than one-third. These results indicate that decisions on surgical management are strongly influenced by the patient's star rating or biological age. If the starring method proves reproducible in other patient groups and settings, it could allow better communication on an important factor in clinical decisions. PMID- 11385091 TI - Why do we have apocrine and sebaceous glands? AB - The secretions of sebaceous and apocrine glands fulfil an important thermoregulatory role in cold-stressed and heat-stressed hunter-gatherers. In hot conditions the secretions emulsify eccrine sweat and thus encourage the formation of a sweat sheet and discourage the formation and loss of sweat drops from the skin. In colder conditions sebum changes its nature and repels rain from skin and hair. PMID- 11385093 TI - Laser treatment of lentiginosis in an Afro-Caribbean. PMID- 11385092 TI - Partial lipodystrophy, polycystic ovary syndrome and proteinuria: a common link to insulin resistance? PMID- 11385094 TI - Paralysis after a diarrhoeal illness. PMID- 11385096 TI - Peritonitis after duodenal stenting. PMID- 11385095 TI - Polymyalgia rheumatica: pitfalls in diagnosis. PMID- 11385097 TI - A learning-disabled woman who had been raped: a multi-agency approach. PMID- 11385098 TI - A tympanitic abdomen. PMID- 11385099 TI - Garibaldi and the surgeons. PMID- 11385100 TI - Colo-broncho-cutaneous fistula complicating traumatic diaphragmatic rupture. PMID- 11385101 TI - Continuous fetal heart rate monitoring. PMID- 11385102 TI - Traditional Chinese medicine and the House of Lords. PMID- 11385104 TI - John Hunter's teachings on gunshot wounds. PMID- 11385105 TI - Localization of oestrogen receptors alpha and beta in human testis. AB - Cellular localization of oestrogen receptor alpha (ERalpha) and beta (ERbeta) proteins were studied in human testis samples using immunohistochemistry, and the expression of the corresponding mRNA was examined with reverse transcription polymerase chain reaction (RT-PCR). Seven men, aged 28-48 years, who underwent diagnostic testicular biopsy because of azoospermia or to give spermatozoa for intracytoplasmic injection for infertility treatment, donated tissue for the study. One of them had anejaculation but normally functioning testes, and one was diagnosed as having Sertoli cell-only syndrome (SCOS). In addition, expression of ERbeta protein was examined in one testis sample obtained from a man undergoing a sex change operation. Strong ERbeta immunoreactivity was detected in the nuclei of spermatogonia, spermatocytes and early developing spermatids. Elongating spermatids, mature spermatozoa, Sertoli and Leydig cells were all negative for ERbeta. The presence of ERbeta protein was confirmed in Western analysis. With RT PCR, both wild-type ERbeta and ERbetacx, the isoform which represses wild-type ER function, were easily detected. In most cases, ERbetacx mRNA was more abundantly expressed than wild-type ERbeta. The patient with SCOS expressed neither ERbeta isoform. Neither ERalpha protein nor ERalpha mRNA was detected in any of the samples. We conclude that in the human testis, ERbeta is likely to be the ER that mediates the effects of oestrogen. PMID- 11385106 TI - MUC1 in normal and impaired spermatogenesis. AB - The MUC1 mucin [also known as episialin, epithelial membrane antigen (EMA) or polymorphic epithelial mucin (PEM)] is a component of the mucosal glycocalyx, contributing to anti-adhesive and protective cell functions. MUC1 has been shown in a variety of epithelial cell types in the reproductive tracts of males and females, but this is the first report of its expression in human testis and non epithelial cells of the germ cell lineage. Analysing 65 testes with normal or impaired spermatogenesis, we identified MUC1 protein in maturing germ cells by immunohistochemistry using the monoclonal antibodies HMFG1, HMFG2 and SM3 binding to different glycosylation variants. MUC1 expression was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot analysis on tissue extracts of human testis, and RT-PCR of selected germ cells after UV laser assisted cell picking. MUC1 glycosylation variants were selectively distributed during normal spermatogenesis. Whereas HMFG1 labelled certain groups of pachytene spermatocytes, HMFG2 labelled only spermatids. Low glycosylated forms of MUC1 mucin, recognized by SM3, were not found. In contrast to its weak expression during normal spermatogenesis, the HMFG1 glycosylation variant accumulated markedly in all spermatocytes showing abnormal or arrested maturation. These results suggest a variable glycosylation of MUC1 mucin in differentiating germ cells, which is aberrant in pathological conditions. PMID- 11385107 TI - Teratozoospermia in mice lacking the transition protein 2 (Tnp2). AB - It is believed that the transition proteins (Tnp1 and Tnp2) participate in the removal of the nucleohistones and in the initial condensation of the spermatid nucleus. Later in spermatogenesis, Tnp1 and Tnp2 are replaced by the protamines 1 and 2. In an effort to elucidate the physiological role of Tnp2, we have disrupted its locus by homologous recombination. Breeding of the Tnp2(-/-) males on different genetic backgrounds revealed normal fertility on the mixed background C57BL/6Jx129/Sv, but total infertility on the inbred 129/Sv background. Light and electron microscopy showed that the germ cells were capable of undergoing chromatin condensation, although many spermatozoa exhibited head abnormalities with acrosomes not attached to the nuclear envelope. Furthermore, migration of Tnp2(-/-) spermatozoa from the uterus into the oviduct was reduced. These results suggest that male infertility of the Tnp2(-/-) mice is a result of sperm head abnormalities and reduced sperm motility. The increased level of the Tnp1 transcript in testes of the Tnp2-deficient mice raises the possibility that a deficiency created through the disruption of the Tnp2 gene can be compensated for by recruitment of the Tnp1. PMID- 11385108 TI - A set of 840 mouse oocyte genes with well-matched human homologues. AB - GenBank contains 14 477 expressed sequence tags (EST) derived from mouse oocyte cDNA libraries: 3499 of these are from two unfertilized oocyte libraries and 10 978 are from two fertilized oocyte libraries. Gene expression profiles were obtained for these libraries by matching library EST to UniGene clusters. The 14 477 EST identified 4226 UNIGENES: These were screened using HomoloGene to identify 1386 homologous UniGene clusters in two other species with one of the matches being human. Within these human matches, 840 encoded named proteins, 223 encoded hypothetical proteins, and 323 encoded clustered EST. The set of named genes provides the first step in establishing a database of genes expressed in mouse oocytes and, by extension, human oocytes. PMID- 11385109 TI - Meta-analysis of gene expression in mouse preimplantation embryo development. AB - Mammalian preimplantation development is characterized by a number of major events. These potentially involve significant but transient changes in early embryonic gene expression. We have undertaken a meta-analysis of gene expression in mouse preimplantation development using a set of 71 346 expressed sequence tags (EST) derived from 15 non-normalized cDNA libraries. These libraries span seven stages of development from the unfertilized oocyte to the blastocyst stage. EST were clustered using UNIGENE: The 71 346 EST identified 11 483 separate genes, of which 1585 are not found elsewhere in the mouse. Aggregate sets of EST for each of the seven stages were analysed for differences in gene expression using Fisher's exact test. This analysis identified 109 genes that were differentially expressed. Some of these genes were associated with degradation of transcripts at the 1-cell stage whereas other genes underwent increased expression at the blastocyst stage. The set of 11 483 genes identified in mouse preimplantation embryo development provides the starting point for the design of DNA microarrays targeted at early mammalian embryogenesis. By anchoring the analysis of mouse preimplantation development in UniGene, it will be possible to identify homologous genes that are likely to be involved in human preimplantation embryo development. PMID- 11385110 TI - Expression of a testis-specific member of the olfactory receptor gene family in human primordial germ cells. AB - Olfactory receptors are G protein-coupled transmembrane receptors. Genes encoding olfactory receptors constitute a large gene family of approximately 1000 total member genes. In mammals, a subset of member genes is specifically expressed in the testis (and not in the olfactory mucosa) and olfactory receptor proteins have been identified in elongated spermatids and mature spermatozoa of dogs. It is postulated that olfactory receptors may recognize signal molecules present in the female genital tract and play a role in chemotaxis of spermatozoa towards the oocyte. In a previous study, we identified 10 cDNA sequences, corresponding to genes specifically expressed in human primordial germ cells (PGC), by differential display. Sequence analysis revealed that one of these sequences appeared to be a member of the olfactory receptor gene family. To investigate this further, we have used degenerate oligonucleotide primers corresponding to conserved amino acid sequences of olfactory receptor proteins to amplify all the olfactory receptor genes expressed in the PGC. Sequence analysis of a total of 30 cloned sequences disclosed that one member gene, which was previously isolated from a human testis cDNA library by others, was also preferentially expressed in our PGC. Our results suggest that specific members of the olfactory receptor gene family may have a function in germ cells in the migratory phase of their life cycle. PMID- 11385111 TI - Co-localization of matrix metalloproteinase-1 and mast cell tryptase in the human uterus. AB - The endometrium displays characteristic cyclical changes involving proliferation and differentiation. The differentiation that takes place requires major tissue remodelling involving the matrix metalloproteinase (MMP) family as key enzymes in this process. Mast cells, containing the tryptase and chymase enzymes that are capable of stimulating the MMP cascade, have been identified in the endometrium, but their role is still unclear. In this study, we observed that the majority of mast cells in the uterus reside in the myometrium and that they co-express mast cell tryptase and MMP-1 in the same intracellular granules. In endometrium exposed to synthetic progestogen via an intrauterine levonorgestrel system a significant increase in mast cells numbers was observed in women experiencing breakthrough bleeding compared to those in women with no reported bleeding. We conclude that mast cells contain MMP-1 and we postulate a potential role for mast cells in breakthrough bleeding. PMID- 11385113 TI - Production of secretory leukocyte protease inhibitor by human amniotic membranes and regulation of its concentration in amniotic fluid. AB - Secretory leukocyte protease inhibitor (SLPI) is a potent inhibitor of human leukocyte elastase. SLPI is a protein found in various human fluids, including parotid secretions, cervical mucus, seminal plasma and ascites. Western blot analysis revealed that SLPI protein is detected as a 12 kDa band in both the amniotic fluid and the amniotic membrane. The amniotic fluid concentrations of SLPI were assayed by enzyme-linked immunosorbent assay. SLPI concentrations in the amniotic fluid of women in the third trimester were higher than those in the second trimester. Immunohistochemistry using an anti-SLPI polyclonal antibody revealed positive staining in epithelial cells in amniotic membranes. Reverse transcription-polymerase chain reaction demonstrated that SLPI transcripts could be detected in the amniotic membranes. To determine the mechanism of SLPI production by amniotic cells, purified amniotic cells were stimulated with various cytokines. Amniotic cells produced SLPI in a dose-dependent manner when stimulated with interleukin (IL)-1alpha, IL-1beta, and tumour necrosis factor alpha. The present findings show that SLPI is produced by the amniotic membranes in response to cytokine concentrations. The SLPI in the amniotic fluid may contribute to immunodefence mechanisms during pregnancy. PMID- 11385112 TI - Progesterone inhibition of functional leptin receptor mRNA expression in human endometrium. AB - Leptin is secreted by adipocytes and regulates appetite through interaction with hypothalamic leptin receptors (OB-R). Leptin is involved in the stimulation of reproductive functions, and local expression of leptin and OB-R in the ovary, oocyte, embryo, and placenta might play a role in early development. The mRNA and protein of the long form leptin receptor (OB-R(L)) but not of leptin are expressed in the human endometrium and the abundance of OB-R mRNA expression varies during the menstrual cycle with a peak in the early secretory phase. We examined the steroidal regulation of OB-R(L) mRNA expression. Northern blot analyses showed that in organ-cultured proliferative endometrial specimens, oestradiol (10(-9) and 10(-8) mol/l) had no acute effect on the OB-R(L) mRNA expression, whereas oestradiol plus progesterone (10(-8), 10(-7) and 10(-6) mol/l) or medroxyprogesterone acetate (10(-8) and 10(-7) mol/l) suppressed the expression by approximately 50%. This progestin-induced suppression was blocked by a concomitant addition of mifepristone. Additionally, incubation of endometrial specimens in the presence of leptin resulted in the phosphorylation of its intracellular target, STAT3 (signal transducer and activator of transcription 3). These results indicate that, in the human endometrium, progestins act via the progesterone receptors to suppress functional OB-R(L) mRNA expression, and may thereby alter the sensitivity of the endometrium to leptin. PMID- 11385114 TI - Human labour is associated with nuclear factor-kappaB activity which mediates cyclo-oxygenase-2 expression and is involved with the 'functional progesterone withdrawal'. AB - Human labour is associated with the up-regulation of prostaglandins within the uterus, synthesized via the type-2 cyclo-oxygenase enzyme (COX-2). These lead to remodelling of the fetal membranes and cervix and to stimulation of myometrial contractions. In the human, the principal source of prostaglandins is the amnion. Progesterone acts to promote myometrial quiescence, and in many species the onset of labour is preceded by withdrawal of progesterone. Humans show no systemic progesterone withdrawal, although biochemical changes within the uterus are similar to those in other species. A mutual negative interaction between the transcription factor nuclear factor (NF)-kappaB and the progesterone receptor (PR) has been reported. Using transient transfections and assays for transcriptional activation and promoter binding, we have shown that there is constitutive activity of NF-kappaB in amnion cells at the time of labour, and that COX-2 expression depends upon NF-kappaB. In cells obtained before labour, in which NF-kappaB activity is low, increasing the concentration of PR represses NF kappaB dependent transcription, while stimulation with IL-1beta both increases NF kappaB activity and represses PR activity. Our data suggest that human labour is associated with constitutive NF-kappaB activity within the amnion, which functions to increase the expression of COX-2 and appears to contribute to the 'functional progesterone withdrawal'. PMID- 11385115 TI - Lipoxygenase gene expression in baboon intrauterine tissues in late pregnancy and parturition. AB - Myometrium (upper and lower segment), cervix, chorion and decidua were obtained at the time of Caesarean hysterectomy in 15 baboons, 10 at varying gestational ages prior to the onset of labour in the last trimester of pregnancy, and five in spontaneous labour at term. Expression of currently recognized human lipoxygenase (LOX) genes was determined using Northern blot analysis relative to each of three housekeeping genes. Signals of similar size to human 5-LOX and human platelet 12 LOX genes were detected in all tissues. Expression of 5-LOX mRNA in the cervix decreased with advancing gestational age. In decidua, expression of 5-LOX mRNA was higher in tissues from animals in labour (compared to those not in labour), whereas in chorion, its expression was lower in tissues from animals in labour. Expression of the platelet 12-LOX gene decreased in chorion with advancing gestational age, and in cervix was lower in tissues from animals in labour. We postulate that the variation in expression of LOX genes may play a role in the onset or promotion of parturition in the baboon. PMID- 11385117 TI - Different models of occupational health service provision and their activity profiles. PMID- 11385116 TI - Evaluation of the tocolytic effect of a selective cyclooxygenase-2 inhibitor in a mouse model of lipopolysaccharide-induced preterm delivery. AB - The inflammatory process is known to cause preterm delivery. Recently, a cyclooxygenase (COX)-2 inhibitor has been developed as an anti-inflammatory drug with few side-effects. We evaluated the COX-2 inhibitor, Celecoxib, for its tocolytic effects and side-effects on dams and pups using a lipopolysaccharide (LPS)-induced preterm delivery mouse model (preterm delivery rates; 95%). With administration of Celecoxib (50, 10, 1 and 0.3 mg/kg), the preterm labour rate was significantly reduced to 18, 30, 36 and 60% respectively. The prostaglandin F(2alpha)(PGF(2alpha)) and PGE(2) concentrations in murine uterine tissue 4 and 10 h after LPS treatment with Celecoxib (10 and 1 mg/kg) were significantly lower than those in the LPS-treated group without CELECOXIB: With administration of 10 or 100 mg/kg Celecoxib, the fetal ductus arteriosus was constricted significantly in preterm and near-term rats, although constriction rates in preterm rats were significantly lower than those in near-term rats. Reproductive and renal functions in offspring whose mothers were treated with LPS and Celecoxib were normal. These data demonstrate that Celecoxib could be used as a new therapy for preterm labour. However, careful attention to constriction of the fetal ductus arteriosus should be given. PMID- 11385118 TI - The quality of inference (based on the 1999 Thackrah Lecture). PMID- 11385119 TI - Clinical and electromyographic examinations of patients with tremor after chronic occupational lead exposure. AB - Tremor is considered to be a clinical sign of patients with chronic lead exposure. However, the type of tremor and its pathophysiological mechanisms are controversial. The aim of this study was to examine the clinical and electromyographic characteristics of tremor in patients with chronic lead exposure. Twenty-three men, 27-49 years old, participated in the study. The tremor activity was detected using surface electrodes from a pair of antagonistic hand muscles. Serum lead concentrations were measured on the day of examination. Our results revealed 12 Hz postural and kinetic tremor, with characteristics of enhanced physiological tremor. Different pathophysiological mechanisms might be responsible for the enhanced physiological tremor of patients with chronic lead exposure. In conclusion, the tremor of patients with chronic lead exposure is an enhanced physiological one. It is probable that this tremor could be influenced by beta-blockers, known to influence the enhanced physiological tremor in anxiety. This will be of benefit for patients disturbed by persistent tremor. PMID- 11385120 TI - The effects of asthma on the quality of life and employment of construction workers. AB - Relatively little is known about the consequences of asthma for patients' work ability. In particular, the relationship between asthma and construction work has not been studied previously. The aim of this study was to determine how asthma affects construction workers' work ability and quality of life, and the most important conditions that cause respiratory symptoms in construction work. A questionnaire was sent to all construction workers examined in the pulmonary clinic of Tampere University Hospital between 1991 and 1995 who were diagnosed as having 'asthma bronchiale' (n = 104). In addition, 206 non-asthmatic construction workers were recruited from the register of the local trade union. Altogether, 73% of the asthmatic construction workers and 70% of the controls completed the questionnaire. Of the asthmatics, 48% were retired, whereas only 24% of the controls were receiving a pension. The asthmatics evaluated their work ability and general health as significantly worse than did the controls. The asthmatics had more limitations in their work and everyday life than the controls. Asthmatic construction workers had often changed work tasks and also occupation because of their illness. Building renovation, cleaning and insulation with mineral fibre materials were the most common causes of respiratory symptoms in the workplace, among both the asthmatics and the controls. Cold air, physical exercise and all types of dust and smoke caused the asthmatics to experience symptoms. It was concluded that asthma limits the work and everyday life of construction workers. Exposure to dusty, non-sensitizing agents is associated with asthmatic symptoms among construction workers. PMID- 11385121 TI - Activity profiles of the occupational health services in a multinational company. AB - The management of the European division of a multinational company was aware of possible differences in the occupational health services (OHS) at their different locations. The objective of this study was to carry out a baseline assessment of these OHS. Structured interviews with representatives of the OHS were conducted at 20 locations in 11 countries. The OHS Recommendation from the International Labour Organization (ILO) was used as a standard for the organization and functions of the OHS. Considerable differences in the activity profiles of the OHS were detected. The inter-enterprise, multidisciplinary OHS spent most of their time on surveillance of workers' health in relation to work and on preventive activities in the working environment. Little time was spent on curative services for individual workers. OHS made up of individual physicians and nurses generally spent much of their time on treatment of occupational and non-occupational diseases. This study has clarified the status of the OHS providers and the potential for improvements in order to meet the needs of the company's locations and to comply more closely with the ILO recommendation. PMID- 11385122 TI - Occupational exposures to metals, solvents and pesticides: recent evidence on male reproductive effects and biological markers. AB - This review is based primarily on the recent epidemiological studies conducted in occupational settings in order to explore the relationship between exposures to chemical agents and the possible effects on male reproductive function. The paper examines evidence of the effects of metals, solvents, pesticides and dioxin. The effects considered are primarily the possible alterations of sperm quality and reduction of fertility. Many studies have identified small groups of workers with exposures to these agents, presenting some alteration in the spermatological or fertility profile, but the results are difficult to replicate in other settings with different individuals and different levels of exposure. From examination of the concentrations of environmental and occupational pollutants in the blood and in the seminal fluid of exposed individuals, it appears that, in general, the concentrations are much lower in the seminal fluid and in sperm cells, making this a less useful marker of exposure. PMID- 11385123 TI - Occupational exposure to airborne proteolytic enzymes and lifestyle risk factors for dental erosion--a cross-sectional study. AB - This study examined the hypothesis that occupational exposure to airborne proteolytic enzymes is associated with dental erosions on the facial surfaces of exposed teeth. Individuals (n = 425) working at a pharmaceutical and biotechnological enterprise (Novozymes A/S) were examined; their mean age was 35 years (range = 18-67 years) and 143 (34%) were women. Two hundred and two of these individuals were newly employed by the company. Occupational exposure was assessed from questionnaire and workplace information. For practical analytical purposes, individuals were categorized as either previously exposed to proteolytic enzymes or not. Information on relevant lifestyle factors and medical history was obtained from a questionnaire. The main effect measure was facial erosion, but lingual erosion indices and the presence of Class V restorations were also considered. The validity of these measures was shown to be very high. Adjusted for potential confounders, there was no association between history of occupational exposure to proteolytic enzymes and prevalent facial or lingual erosion. With respect to prevalence of Class V restorations, the association was significant. The present study did not support directly our primary hypothesis that occupational exposure to airborne proteolytic enzymes is associated with dental erosions on the facial surfaces of exposed teeth. However, the results indicate that exposure to proteolytic enzymes may lead to pronounced tooth substance loss, demanding treatment. PMID- 11385124 TI - Environmental and biological monitoring of traffic wardens from the city of Rome. AB - A molecular epidemiological study on Roman policemen is ongoing. The results of a first assessment of the occupational exposure to aromatic compounds of 66 subjects engaged in traffic control and of 33 office workers are presented in this paper. Passive personal samplers and urinary biomarkers were used to assess exposure to benzene and polycyclic hydrocarbons during work shifts. The results obtained indicate that benzene exposure in outdoor workers is about twice as high as in office workers (geometric mean 7.5 and 3.4 micrograms/m3, respectively). The distribution of individual exposure values was asymmetrical and skewed toward higher values, especially among traffic wardens. Environmental benzene levels recorded by municipal monitoring stations during work shifts (geometric mean 11.2 micrograms/m3) were in the first instance comparable to or greater than individual exposure values. However, several outlier values were observed among personal data that greatly exceeded average environmental benzene concentrations. Among the exposure biomarkers investigated, only blood benzene correlated to some extent with previous exposure to benzene, while a seasonal variation in the excretion of 1-hydroxypyrene and trans-muconic acid was observed in both study groups. In conclusion, these results suggest that outdoor work gives a greater contribution than indoor activities to benzene exposure of Roman citizens. Moreover, relatively high-level exposures can be experienced by outdoor workers, even in the absence of large-scale pollution episodes. PMID- 11385125 TI - SWORD '99: surveillance of work-related and occupational respiratory disease in the UK. AB - Systematic reports from chest and occupational physicians under the SWORD and OPRA (Occupational Physicians Reporting Activity) surveillance schemes continue to provide a picture of the incidence of occupational respiratory disease in the UK. An estimated total of 4393 incident cases (comprising 4530 diagnoses) were reported during the 1999 calendar year, an increase of 1427 cases over the previous year. Benign pleural disease was the single most frequently reported condition (28% of all diagnoses reported). Occupational asthma cases (1168; 26%) remained high, as did mesothelioma (1032; 23%). Analysis of trends over the past 8 years shows an increase in mesothelioma cases, but little change in asthma. The annual incidence per 100,000 employed people, 1996-1999, for mesothelioma, lung cancer and pneumoconiosis was high amongst construction workers (28.7), miners and quarrymen (26.5), woodworkers (18.9) and gas, coal and chemical workers (15.2). Trends in mesothelioma incidence by birth cohort continue to show an increase in construction workers and a continuing decline in shipyard and insulation workers. The relative proportion of pneumoconiosis cases attributed to coal mining has fallen steadily in workers born since approximately 1920 and most cases are now in men who have been employed in quarrying and rock drilling. PMID- 11385127 TI - Health, Environment and Work. Raymond Agius. http://www.agius.com/hew/index.htm. PMID- 11385126 TI - Perceived work ability of home care workers in relation to individual and work related factors in different age groups. AB - This study analysed the relationship between age and items of the Work Ability Index (WAI) among 19- to 62-year-old female home care workers (n = 636). The first significant decrease in work ability occurred between the ages of 40 and 44 years, and a second, sharper decrease occurred after 55 years of age. The subjects' physical work ability deteriorated as early as 35 years of age. In addition, current work ability, number of diagnosed diseases and work impairment due to diseases proved to be the most age-sensitive measures of work ability. The subjects who perceived their health status as poor had the highest risk for poor work ability. Moreover, the age-adjusted odds ratio indicated that the WAI was strongly associated with age and musculoskeletal and psychosomatic symptoms. Ergonomics, possibilities to control one's own work, time pressure and management were the features of work that predicted work ability. The study demonstrated the need to promote the work ability of home care workers early in working life. Among both older and younger workers, preventive measures should focus on the above-mentioned factors. PMID- 11385128 TI - Putting growth factors into context. PMID- 11385129 TI - The use of glyburide in gestational diabetes--an ideal example of "bench to bedside". PMID- 11385130 TI - Neurobiology of hypoxic-ischemic injury in the developing brain. AB - Hypoxic ischemia is a common cause of damage to the fetal and neonatal brain. Although systemic and cerebrovascular physiologic factors play an important role in the initial phases of hypoxic-ischemic injuries, the intrinsic vulnerability of specific cell types and systems in the developing brain may be more important in determining the final pattern of damage and functional disability. Excitotoxicity, a term applied to the death of neurons and certain other cells caused by overstimulation of excitatory, mainly glutamate, neurotransmitter receptors, plays a critical role in these processes. Selected neuronal circuits as well as certain populations of glia such as immature periventricular oligodendroglia may die from excitotoxicity triggered by hypoxic ischemia. These patterns of neuropathologic vulnerability are associated with clinical syndromes of neurologic disability such as the extrapyramidal and spastic diplegia forms of cerebral palsy. The cascade of biochemical and histopathologic events triggered by hypoxic ischemia can extend for days to weeks after the insult is triggered, creating the potential for therapeutic interventions. PMID- 11385131 TI - Prevention of fetal malformation with antioxidants in diabetic pregnancy. PMID- 11385132 TI - Maternal vitamin A administration and the fetal ductus arteriosus. PMID- 11385133 TI - The effect of vitamin A on contraction of the ductus arteriosus in fetal rat. AB - Endogenous retinoic acid may play a role in inducing smooth muscle differentiation in the fetal ductus arteriosus. Maternal administration of retinoic acid may accelerate the process. This study was designed to investigate the effect of vitamin A on developmental changes in the contractile system of the ductus. Vitamin A was injected into pregnant rats and the ductus was isolated from the fetus at 19, 20, or 21 d of gestation. The fetus at 19 d of gestation served as a model of the preterm fetus. The force of contraction and [Ca]i were measured. Membrane depolarization caused by high KCl induced ductal contraction in all age groups studied. In the 19-d fetus, O2 did not cause significant contraction or changes in [Ca]i in the control group, but it did induce a significant contraction and increases in [Ca]i in the vitamin A-treated group. In the 20- and 21-d fetuses, 5% O2-induced contraction in the vitamin A-treated group was significantly greater than in the control group. In the 19-d fetus, noradrenaline-induced contraction and increases in [Ca]i, indicators of the size of the intracellular Ca pool, were observed and they were similar in the control group and in the vitamin A-treated group. These data suggest that 1) in the preterm fetus, the contractile system, including membrane depolarization, [Ca]i increase, and its activation of contractile proteins, is already functioning, but the O2-sensing mechanism is underdeveloped, 2) vitamin A accelerates the development of the O2-sensing mechanism of the ductus arteriosus. PMID- 11385134 TI - Combined treatment with vitamin E and vitamin C decreases oxidative stress and improves fetal outcome in experimental diabetic pregnancy. AB - The aim was to investigate whether dietary supplementation of a combination of the two antioxidants, vitamin E and vitamin C, would protect the fetus in diabetic rat pregnancy at a lower dose than previously used. Normal and streptozotocin-induced diabetic rats were mated and given standard food or food supplemented with either 0.5% vitamin E + 1% vitamin C or 2% vitamin E + 4% vitamin C. At gestational d 20, gross morphology and weights of fetuses were evaluated. Vitamins E and C and thiobarbituric acid reactive substances were measured in maternal and fetal compartments. In addition, protein carbonylation was estimated in fetal liver. Maternal diabetes increased the rate of malformation and resorption in the offspring. High-dose antioxidant supplementation decreased fetal dysmorphogenesis to near normal levels. The low dose group showed malformations and resorptions at an intermediate rate between the untreated and the high-dose groups. Thiobarbituric acid reactive substances were increased in fetal livers of diabetic rats and reduced to normal levels already by low-dose antioxidative treatment. Protein carbonylation rate was also increased in fetal liver of diabetic rats; it was normalized by high-dose treatment but only partially reduced by low-dose antioxidants. We conclude that combined antioxidative treatment with vitamins E and C decreases fetal malformation rate and diminishes oxygen radical-related tissue damage. However, no synergistic effect between the two antioxidants was noted, a result that may influence future attempts to design antiteratogenic treatments in diabetic pregnancy. Oxidatively modified proteins may be teratogenically important mediators in diabetic embryopathy. PMID- 11385135 TI - Tall or short? Twenty years after preeclampsia exposure in utero: comparisons of final height, body mass index, waist-to-hip ratio, and age at menarche among women, exposed and unexposed to preeclampsia during fetal life. AB - Women exposed to preeclampsia during fetal life have lower risk of breast cancer, compared with unexposed women, possibly through fetal programming. Hypothetically, preeclampsia exposure could affect well-known risk factors for breast cancer, such as pubertal development or adult anthropometry. Women born in a defined geographic area of Sweden from 1973 through 1978, with verified preeclampsia exposure (n = 230) and nonexposure (n = 359) during fetal life, answered questions about anthropometric measures, smoking, parity, and age at menarche in a telephone interview in early adulthood. Compared with unexposed offspring, female offspring of women who had preeclampsia were lighter and shorter for gestational age, but in young adulthood there were no differences in height, body mass index, waist-to-hip ratio, or age at menarche. When analyzing the effects of other maternal and fetal characteristics, the results indicate that approximately 50% of the variance in final height was explained by parental heights and birth length for gestational age. Young-adult body mass index was weakly associated with maternal body mass index, maternal smoking, and birth weight for gestational age, which together explained 12% of the variance. Neither of the assessed maternal or fetal characteristics were significantly associated with age at menarche or waist-to-hip ratio. These data indicate that neither adult anthropometry nor age at menarche is in the causal pathway between intrauterine preeclampsia exposure and the reduced risk of breast cancer. PMID- 11385136 TI - Utility and limitations of near-infrared spectroscopy during cardiopulmonary bypass in a piglet model. AB - Near-infrared spectroscopy assessment of cytochrome oxygenation could be a valuable technique to monitor cerebral intraneuronal oxygen delivery during cardiopulmonary bypass. However, the validity of the cytochrome signal has been questioned as it could easily be overwhelmed by the Hb signal. Five- to six-week old control piglets (n = 5) underwent cardiopulmonary bypass at 37 degrees C. Study animals (n = 10) received 100 mg/kg of sodium cyanide to uncouple cytochrome from HB: Hematocrit was then decreased in steps of 5% from 35 to 5% with crystalloid hemodilution. In study piglets, the initiation of cardiopulmonary bypass was associated with oxygenated Hb increasing from 0 to 62.9 +/- 25.6 microM times the differential path-length factor, and oxidized cytochrome a,a3 increasing to 1.9 +/- 1.8 microM times the differential path length factor. Cyanide caused oxygenated Hb to increase further to 132.3 +/- 48.9 microM times the differential path-length factor, and oxidized cytochrome c decreased to -7.0 +/- 2.6 microM times the differential path-length factor as anticipated, confirming uncoupling of electron transport. However, hemodilution subsequently resulted in linear decreases in oxidized cytochrome a,a3 (F = 8.57, p < 0.001) suggesting important cross-talk between the Hb and cytochrome signals as cytochrome is only intracellular. In control piglets, tissue oxygenation index showed a positive correlation with pump flow (r = 0.986, p = 0.013). The cytochrome signal as presently measured by near-infrared spectroscopy is highly dependent on hematocrit. PMID- 11385137 TI - Antenatal dexamethasone administration impairs normal postnatal lung growth in rats. AB - Our purpose was to determine the influences of antenatal dexamethasone administration on neonatal lung development in rats. Dexamethasone (0.4 mg/kg maternal body weight per day) was administered i.p. on the 21st d (group 1) or on the 20th and 21st d (group 2) of gestation in Sprague Dawley rats with timed pregnancies. After natural deliveries, the lungs of the pups 1-60 d of age were removed and processed for morphometric analysis. In 60-d-old pups, groups 1 and 2 both showed a lower numerical density of alveoli and a larger mean alveolar radius than control pups. Antenatal administration of dexamethasone to rats impairs the normal postnatal lung growth. Some aspects of the use of antenatal glucocorticoid therapy in humans may require reconsideration. PMID- 11385138 TI - Development of glucocorticoid-responsiveness in mouse intestine. AB - There are conflicting data from human studies regarding the ability of exogenous glucocorticoids to stimulate maturation of the small intestine. The discrepancies may relate to differences in hormone doses and age administered. To explore this general concept, we have used a mouse model to determine intestinal responsiveness to dexamethasone (DEX) at various times during development. We first showed that trehalase mRNA is a sensitive marker of intestinal maturation in the mouse; being undetectable (by Northern blotting) in the prenatal period, expressed at low levels during the first 2 postnatal weeks and then displaying a marked increase in the 3rd postnatal week. DEX was unable to elicit detectable trehalase mRNA in fetal mice, but caused significant increases in the postnatal period. The use of a range of DEX doses (0.0125-2.5 nmol/g BW per day) established that there is no change in sensitivity between the 1st and 2nd postnatal weeks, but there is a significant increase in maximal responsiveness of trehalase mRNA to the hormone. Similar results were obtained when sucrase isomaltase mRNA was assayed in the same animals. Thus, in this rodent model, there appears to be at least three phases in the DEX responsiveness of the developing intestine: an early phase (prenatal) when DEX is unable to elicit intestinal maturation; then a phase (first postnatal week) of modest responsiveness; then a transition to increased responsiveness. These findings point to the need for careful attention to dose and age in analyses of glucocorticoid effects in human infants. PMID- 11385139 TI - Developmental expression of PEPT1 and PEPT2 in rat small intestine, colon, and kidney. AB - Mammalian peptide transporters (PEPT1 and PEPT2) play a pivotal role in the absorption of small peptides from the intestine and kidney, respectively, and in the disposition and targeting of peptide or mimetic drugs. However, there are few reports on the molecular basis of their regulation, especially in the young. The aim of this study was to determine the developmental expression of intestinal and renal oligopeptide transporters in rats from embryonic to adult ages. Intestinal segments were collected (i.e. duodenum, jejunum, ileum, and colon) along with whole kidney, and their mRNA and protein levels were measured. Expression levels of PEPT1 were maximal 3-5 d after birth in the duodenum, jejunum, and ileum, and then declined rapidly. Expression was increased transiently at d 24, most notably in the ileum. Adult protein levels were approximately 70% of that observed on d 3 5. Significant PEPT1 expression was observed in colon during the first week of life, but levels were undetectable shortly thereafter through adulthood. PEPT1 and PEPT2 expression is less regulated in rat kidney and more pronounced in older animals. Peptide transporters were also present as early as d 20 of fetal life for all tissues tested. These results are unique in providing the developmental expression of peptide transporter mRNA and protein in distinct regions of the small intestine, colon, and kidney in rat. Our findings suggest that intestinal expression of PEPT1 is induced postpartum, possibly by suckling, and again at the time of weaning, and that the colon may participate in peptide transport early in life. PMID- 11385140 TI - Influence of metabolic control on growth in homocystinuria due to cystathionine B synthase deficiency. AB - The etiology of the tall stature almost invariably seen in homocystinuric patients is not known. The effect of metabolic control and the role of the GH-IGF system on growth were investigated in 10 patients with homocystinuria. There was a direct correlation between the plasma free homocyst(e)ine and growth velocity SD scores in 18 patient years (r, 0.46; p < 0.05). Plasma 2-y cumulative free homocyst(e)ine and height SD scores were directly correlated (r, 0.82; p < 0.01). Growth velocity SD scores were lower in patients with optimal metabolic control than in those with poorer control (-0.10 +/- 0.65 versus 0.95 +/- 0.68, p < 0.01). Height SD scores were also lower in the optimally controlled group (-0.01 +/- 0.81 versus 1.73 +/- 0.88, p < 0.05). GH and GH-related peptides did not deviate significantly from the reference ranges. These findings suggest that overgrowth is directly mediated by homocysteine, that the GH-IGF axis is not involved, and that it may be prevented by optimal metabolic control. PMID- 11385141 TI - Effect of extreme hypercapnia on hypoxic-ischemic brain damage in the immature rat. AB - To ascertain the effect of extreme hypercapnia on perinatal hypoxic-ischemic brain damage, 7-d-postnatal rats were exposed to unilateral common carotid artery occlusion followed by hypoxia with 8% oxygen combined with 3, 12, or 15% carbon dioxide (CO2) for 2 h at 37 degrees C. Survivors underwent neuropathologic examination at 30 d of postnatal age, and their brains were characterized as follows: 0 = normal; 1 = mild atrophy; 2 = moderate atrophy; 3 = cystic infarct with external dimensions <3 mm; and 4 = cystic infarct with external dimensions >3 mm. The width of the cerebral hemisphere ipsilateral to the carotid artery occlusion also was determined on a posterior coronal section and compared with that of the contralateral hemisphere to ascertain the severity of cerebral atrophy/cavitation. CO2 tensions averaged 5.08, 11.1, and 13.2 kPa in the 3, 12, and 15% CO2-exposed animals, respectively, during hypoxia-ischemia (HI). Neuropathologic results showed that immature rats exposed to 3 and 12% CO2 had similar severities of brain damage. In contrast, rat pups exposed to HI combined with 15% CO2 were significantly more brain damaged than littermates exposed to 3% CO2. Specifically, eight of 14 animals exposed to 15% CO2 showed cystic infarcts (grades 3 and 4), whereas none of 14 littermates exposed to 3% CO2 developed cystic infarcts (p < 0.01). Analyses of coronal width ratios at each CO2 exposure provided results comparable with those of the gross neuropathology scores. Cerebral blood flow (CBF), measured at 90 min of HI, was lowest in those immature rats exposed to 15% CO2 compared with control (p = 0.04), with higher values in those rat pups exposed to 3 and 12% CO2. The findings indicate that 7-d-postnatal rats exposed to HI with superimposed 12% CO2 are neither less nor more brain damaged than littermates exposed to 3% CO(2) (normocapnia). In contrast, animals exposed to 15% CO2 are the most brain damaged of the three groups. Presumably, extreme hypercapnia produces more severe cardiovascular depression than is seen in animals subjected to lesser degrees of hypercapnia; the cardiovascular depression, in turn, leads to greater cerebral ischemia and ultimate brain damage. PMID- 11385142 TI - Platelet-activating factor antagonist BN 50730 attenuates hypoxic-ischemic brain injury in neonatal rats. AB - Platelet-activating factor (PAF) is a lipid derived from breakdown of cell membranes that is postulated to be a mediator of cerebral ischemic injury. PAF regulates CNS gene transcription via intracellular binding sites. To test the hypothesis that PAF mediates CNS injury in part by modulating gene transcription, we evaluated the neuroprotective efficacy of the drug BN 50730, an antagonist of the intracellular (microsomal) CNS PAF binding site, in the neonatal rat model of unilateral cerebral hypoxia-ischemia. Seven-day-old rats underwent right carotid ligation followed by a 2.5-h exposure to 8% O(2), and were then treated with BN 50730 (2.5 or 25 mg/kg per dose) or vehicle, at 0 and 2 h after the end of hypoxia. Ipsilateral cortical, striatal, and hippocampal damage was quantitated either 5 d later, or at 5 wk after the insult. Treatment with BN 50730 resulted in approximately 60- 80% reduction in ipsilateral tissue loss at both times. Learning and memory were evaluated 5 wk after insult using the Morris Watermaze place navigation task. Severity of cortical and striatal damage correlated significantly with learning and memory deficits. These results support the hypothesis that PAF is a pathogenetic mediator of hypoxic-ischemic damage in the immature brain. Accumulating evidence suggests that PAF mediates its deleterious effects in the immature CNS via multiple mechanisms. PMID- 11385143 TI - Impaired early neurologic outcome in newborn piglets reoxygenated with 100% oxygen compared with room air after pneumothorax-induced asphyxia. AB - Birth asphyxia is a serious problem worldwide, resulting in 1 million deaths and an equal number of neurologic sequelae annually. It is therefore important to develop new and better ways to treat asphyxia. In the present study we tested the effects of reoxygenation with room air or with 100% oxygen (O2) after experimental pneumothorax-induced asphyxia on the blood oxidative stress indicators, early neurologic outcome, and cerebral histopathology of newborn piglets. Twenty-six animals were studied in three experimental groups: 1) sham operated animals (SHAM, n = 6), 2) animals reoxygenated with room air after pneumothorax (R21, n = 10), and 3) animals reoxygenated with 100% O2 after pneumothorax (R100, n = 10). In groups R21 and R100, asphyxia was induced under anesthesia with bilateral intrapleural room air insufflation. Gasping, bradyarrhythmia, arterial hypotension, hypoxemia, hypercarbia, and combined acidosis occurred 62 +/- 6 min (R21) or 65 +/- 7 min (R100; mean +/- SD) after the start of the experiments; then pneumothorax was relieved, and a 10-min reoxygenation period was started with mechanical ventilation with room air (R21) or with 100% O2 (R100). The newborn piglets then breathed room air spontaneously during the next 3 h. Blood oxidative stress indicators (oxidized and reduced glutathione, plasma Hb, and malondialdehyde concentrations) were measured at different stages of the experiments. Early neurologic outcome examinations (neurologic score of 20 indicates normal, 5 indicates brain-dead) were performed at the end of the study. The brains were next fixed, and various regions were stained for cerebral histopathology. In the SHAM group, the blood gas and acid base status differed significantly from those measured in groups R21 and R100. In group R100, arterial PO2 was significantly higher after 5 (13.8 +/- 5.6 kPa) and 10 min (13.2 +/- 6.3 kPa) of reoxygenation than in group R21 (8.7 +/- 2.8 kPa and 9.2 +/- 3.1 kPa). The levels of all oxidative stress indicators remained unchanged in the study groups (SHAM, R21, and R100). The neurologic examination score in the SHAM group was 18 +/- 0, in group R21 it was 13.5 +/- 3.1, and in group R100 it was 9.5 +/- 4.1 (significant differences between SHAM and R21 or R100, and between R21 and R100). Cerebral histopathology revealed marked damage of similar severity in both asphyxiated groups. We conclude that the blood oxidative stress indicators and cerebral histopathology did not differ significantly after a 10-min period of reoxygenation with room air or with 100% O2 after pneumothorax-induced asphyxia, but reoxygenation with 100% O2 might impair the early neurologic outcome of newborn piglets. PMID- 11385144 TI - Meconium induces expression of inducible NO synthase and activation of NF-kappaB in rat alveolar macrophages. AB - Meconium aspiration causes intensive inflammatory reactions in the lungs, and may lead to neonatal respiratory disorder. Infiltrated inflammatory cells, particularly macrophages, play an important role in such an inflammation. A rat alveolar macrophage cell line (ATCC8383) was exposed to meconium alone or in combination with dexamethasone, budesonide, or interferon-gamma. Nitric oxide (NO) accumulation in the supernatant of the cell culture was detected by Griess reaction, and mRNA of inducible NO synthase (iNOS) expression was detected by reverse transcriptase-PCR. Nuclear factor-kappa B was analyzed by electrophoretic mobility shift assay, and iNOS location and nuclear factor-kappa B transactivation were determined by immunostaining. Our results showed that meconium was capable of inducing production of NO and expression of iNOS in alveolar macrophages in a dose- (1-25 mg/mL, p < 0.05) and time- (4-48 h, p < 0.05) dependent manner. This capability of meconium could be further enhanced in the presence of interferon-gamma (100 IU/mL, p < 0.05). Budesonide (10(-4)-10( 10) M) or dexamethasone (10(-4)-10(-6) M) effectively inhibited the meconium induced NO production (p < 0.05). Using the protein synthesis inhibitor cycloheximide, we demonstrated that meconium directly induced iNOS in macrophages. Furthermore, meconium also triggered nuclear factor-kappa B activation, a mechanism possibly responsible for the iNOS expression. Our findings suggest that meconium is a potent inflammatory stimulus, resulting in iNOS expression, leading to overproduction of NO from the macrophages, which may be of pathogenic importance in meconium aspiration syndrome. In vitro steroids down-regulated the iNOS expression, thus suggesting a potential to down-regulate NO-mediated inflammation in neonates with meconium aspiration syndrome. PMID- 11385145 TI - Responses of fetal ovine systemic and umbilical arteries to angiotensin II. AB - Angiotensin II (ANG II) contracts umbilical arteries and has been hypothesized to regulate fetal blood pressure primarily by altering umbilical vascular resistance. To determine whether systemic arteries in term fetal sheep are sensitive to ANG II, isometric contraction of endothelium-intact isolated fetal renal, mesenteric, and umbilical arteries in response to ANG II was studied. ANG II (10(-7) M) elicited contractile responses in all three vessels (43 +/- 8%, 99 +/- 21%, and 105 +/- 5% of the maximal response seen with 90 mM KCl for renal, mesenteric, and umbilical arteries, respectively). The time course of the contractile responses differed among the vessels: renal and mesenteric arteries exhibited rapid transient contraction followed by relaxation, whereas umbilical artery displayed a more slowly developing but sustained contraction (1 +/- 0%, 3 +/- 1%,and 93 +/- 4% of maximal contractile response at 5 min, for renal, mesenteric, and umbilical arteries, respectively). The AT1 receptor antagonist, losartan (10(-6) M), abolished contractile responses in renal and mesenteric arteries but only slowed the contraction in umbilical artery in response to ANG II and had no effect on maximal tension. AT2 receptor blockade (PD 123319, 10(-7) M) had no significant effect on the response to ANG II in any vessel. Indomethacin (10(-6) M) significantly potentiated contraction to ANG II in renal and mesenteric but not umbilical arteries. Northern and Western blot analyses demonstrated the presence of AT1 mRNA and protein in all three vessels. Immunostaining for the AT1 receptor was present in endothelium and the tunica media. These findings demonstrate the AT1 receptor is present and functionally active in fetal systemic arteries and are consistent with previous findings that the umbilical circulation displays a greater responsiveness to ANG II than the systemic vasculature. PMID- 11385146 TI - Hydrogen peroxide production in leukocytes during cerebral hypoxia and reoxygenation with 100% or 21% oxygen in newborn piglets. AB - The aim of this study was to investigate whether reoxygenation with 21% O2 rather than 100% O2 results in reduced hydrogen peroxide (H2O2) concentrations in neutrophils (PMN). Piglets (2-4 d old) exposed to severe hypoxia (inspired fraction of oxygen, 0.08) were randomized to resuscitation with 21 (n = 13) or 100% O2 (n = 12). Five animals served as controls. H2O2 concentrations in PMN in terms of rhodamine 123 (Rho 123) fluorescence intensity from arterial and superior sagittal sinus blood were quantified by flow cytometry. Laser Doppler flowmetry (LDF) was used to assess cortical blood perfusion. During hypoxia, Rho 123 increased in arterial PMN in both study groups by 15 and 32%, respectively (p < 0.05). In cerebral venous PMN, the increase was less dominant (p = 0.06). Reoxygenation with 100 or 21% O2 had no different effect on Rho 123 in arterial PMN. In cerebral venous PMN, Rho 123 was approximately 40% higher after 60 min and 30% higher after 120 min compared with corresponding data in the 21% O2 group (p < 0.05), which were close to baseline levels. Further, O2 treatment in both groups induced PMN accumulation in arterial blood (p < 0.05). Laser Doppler flowmetry signals increased during transient hypoxia (p < 0.0001 compared with baseline) and were normalized after reoxygenation in both study groups. In conclusion, arterial and cerebral venous H2O2 concentration in PMN tended to increase during hypoxia. During reoxygenation, H2O2 concentration in PMN in the cerebral circulation was low with 21% O2 but remained high with 100% O2 ventilation. We speculate that oxygen should be reintroduced with more caution during neonatal resuscitation. PMID- 11385147 TI - Biologic role of fetal lung fibroblast triglycerides as antioxidants. AB - The pulmonary response to hyperoxia is highly variable, depending on such seemingly disparate biologic factors as gestational age, sex, hormonal milieu, and nutritional status. Descriptively, the magnitude and direction of these biologic differences in response to hyperoxia correlate with the triglyceride content of developing fetal rat lung fibroblasts (FRLFs). Mechanistically, these same factors affect the triglyceride content of FRLFs, e.g. d 21 FRLFs contain more triglyceride than d 18 FRLFs; female FRLFs contain more triglyceride than male FRLFs (d 20); dexamethasone increases FRLF triglyceride content, dihydrotestosterone decreases it; nutritionally, exposure of FRLFs to graded amounts of serum triglyceride (0%, 2%, 10%, 20%) results in increased intracellular FRLF triglyceride content. To test the hypothesis that these biologic differences in intracellular triglyceride content may account for differences in the cytoprotection of lung fibroblasts against oxidant injury, fibroblast cultures representing each of these biologic groups were challenged with graded doses of the reactive oxygen species hydrogen peroxide (0.1-1.0 mM for 5 min). The number of surviving cells and their antioxidant status, as measured by lipid peroxidation and glutathione content of the surviving cells, were determined. We found that in response to hydrogen peroxide 1) d 21 FRLFs were more resistant than d 18 FRLFs; 2) female FRLFs were more resistant than male FRLFs; 3) dexamethasone-treated FRLFs were more resistant than dihydrotestosterone-treated fibroblasts; 4) fibroblasts fed increasing amounts of serum triglycerides were increasingly resistant to hydrogen peroxide; 5) cell survival in different serum triglyceride- and hormone-treated groups was not related to the antioxidant status as measured by glutathione content. These data are consistent with the hypothesized role of FRLF triglycerides as antioxidants. PMID- 11385148 TI - Femoral arterial puncture management after percutaneous coronary procedures: a comparison of clinical outcomes and patient satisfaction between manual compression and two different vascular closure devices. AB - BACKGROUND: Vascular access site management is crucial to safe, efficient and comfortable diagnostic or interventional transfemoral percutaneous coronary procedures. Two new femoral access site closure devices, Perclose and Angio-Seal , have been proposed as alternative methods to manual compression (MC). We compared these two devices and tested them in reference to standard MC for safety, effectiveness and patient preference. METHODS: Prospective demographic, peri-procedural, and late follow-up data for 1,500 patients undergoing percutaneous coronary procedures were collected from patients receiving femoral artery closure by MC (n = 469), Perclose (n = 492), or Angio-Seal (n = 539). Peri procedural, post-procedural, and post-hospitalization endpoints were: 1) safety of closure method; 2) efficacy of closure method; and 3) patient satisfaction. RESULTS: Patients treated with Angio-Seal experienced shorter times to hemostasis (p < 0.0001, diagnostic and interventional) and ambulation (diagnostic, p = 0.05; interventional, p < 0.0001) than those treated with Perclose. Those treated with Perclose experienced greater access site complications (Perclose vs. Angio-Seal, p = 0.008; Perclose vs. MC, p = 0.06). Patients treated with Angio-Seal reported greater overall satisfaction, better wound healing and lower discomfort (each vs. Perclose or vs. MC, all p < or = 0.0001). For diagnostic cath only, median post procedural length of stay was reduced by Angio-Seal (Angio-Seal vs. MC, p < 0.0001; Angio-Seal vs. Perclose, p = 0.009). No difference was seen in length of stay for interventional cases. CONCLUSIONS: Overall, Angio-Seal performed better than Perclose or MC in reducing time to ambulation and length of stay among patients undergoing diagnostic procedures. There was a higher rate of successful deployment and shorter time to hemostasis for Angio-Seal, and this was accomplished with no increase in bleeding complications throughout the follow-up. Additionally, Angio-Seal performed better than Perclose in exhibiting a superior 30-day patient satisfaction and patient assessment of wound healing with less discomfort. PMID- 11385149 TI - Percutaneous intervention for symptomatic vertebral artery stenosis using coronary stents. AB - BACKGROUND: There is very limited experience with percutaneous treatment of symptomatic vertebral artery disease. Angioplasty and stenting for vertebral artery stenosis are still evolving and have generally been performed for asymptomatic disease. We performed vertebral artery stenting in 12 patients with vertebrobasilar transient ischemic attacks and present our short- and intermediate-term results. METHODS: A total of 12 lesions affecting the vertebral artery were treated by coronary stent placement. The mean age was 72 +/- 8 years and 83% were males (10 males, 2 females). Baseline characteristics included hypertension (11/12); hypercholesterolemia (8/12); coronary artery disease (8/12); and diabetes (5/12). Mean lesion length was 8.6 +/- 2.7 mm, mean calipered stenosis was 78 +/- 8%, and mean arterial diameter was 4.1 +/- 0.3 mm. All patients were symptomatic, fulfilling our criteria for vertebral artery angioplasty. All patients were followed for at least 6 months after treatment. RESULTS: All 12 lesions were successfully stented, with a mean residual stenosis of 11 +/- 6%. Clinical follow-up showed resolution or improvement of symptoms in all patients. One patient had symptomatic restenosis seven months after the initial procedure requiring repeat angioplasty. CONCLUSIONS: Stent placement for symptomatic stenosis involving the vertebral artery is safe and effective for alleviating symptoms of vertebrobasilar ischemia. Coronary stents appear to be well suited to treat atherosclerotic lesions of the vertebral artery. PMID- 11385151 TI - How late is too late (for infarct angioplasty)? PMID- 11385150 TI - Primary angioplasty versus no reperfusion therapy in patients with acute myocardial infarction and a pre-hospital delay of > 12-24 hours: results from the pooled data of the maximal individual therapy in acute myocardial infarction (MITRA) registry and the myocardial infarction registry (MIR). AB - OBJECTIVE: In patients with acute myocardial infarction (AMI), treatment with thrombolysis is superior to no reperfusion therapy only up to 12 hours after the onset of symptoms. There are no data addressing whether this time limit is also justified for treatment with primary angioplasty. DESIGN: The pooled data of two German ST-segment elevation AMI registries, the Maximal Individual Therapy in Acute Myocardial Infarction (MITRA) study and the Myocardial Infarction Registry (MIR), were analyzed. PATIENTS: Out of 22,749 patients, eight hundred and forty eight with a pre-hospital delay of > 12 hours and < or = 24 hours were treated with either primary angioplasty (94/848; 11.1%) or no reperfusion therapy (754/848; 88.9%). RESULTS: Patients treated with primary angioplasty were 10 years younger (59 years versus 69 years; p = 0.001), more often male [72.3% versus 59.9%; odds ratio (OR) = 0.57; 95% confidence interval (CI) = 0.36-0.92] and less likely to be diabetics (17% versus 27.2%; OR = 0.55; 95% CI = 0.31 0.97). Hospital mortality was 8.5% in patients treated with primary angioplasty compared to 17.1% in patients with no reperfusion therapy (OR = 0.45; 95% CI = 0.21-0.95; p = 0.033) and the combined endpoint (death, reinfarction or stroke) occurred significantly less often (11.7% versus 20.3%; OR = 0.52; 95% CI =0.27-1; p = 0.045). However, multiple logistic regression showed only a non-significant trend for lower mortality (OR = 0.54; 95% CI =0.20-1.23) and the combined endpoint (OR = 0.65; 95% CI = 0.29-1.31) in patients treated with primary angioplasty. CONCLUSIONS: These data show the possibility of a benefit of primary angioplasty over conservative treatment in patients with pre-hospital delays of > 12 up to 24 hours, although multiple logistic regression analysis failed to find significant differences between treatments. This might be due to inadequate study power or a selection bias. These findings encourage further investigation of this subject. PMID- 11385152 TI - Outcomes following extracranial carotid artery stenting in high-risk patients. AB - BACKGROUND: Carotid artery angioplasty and stenting has become a viable alternative to carotid endarterectomy (CEA), especially for patients considered at high risk for post-operative complications. This study investigated the feasibility, safety and long-term outcome of carotid artery stenting (CAS) in high-risk patients. METHODS: From July 1995 to November 2000, sixty-two consecutive patients considered to be at high risk for post-operative complications of CEA were followed prospectively after undergoing extracranial CAS procedures. RESULTS: Sixty-two patients [37 men (60%) and 25 women (40%)] underwent a total of 69 CAS procedures. The mean age was 67 +/- 9 years (range, 32-89 years). Comorbid conditions included hypertension in 95% and severe coronary artery disease in 58%. Sixteen patients (26%) had a previous ipsilateral CEA, twenty-one percent had a history of neck radiation and 32% had a history of significant contralateral carotid artery disease. Fifty-two patients (84%) were symptomatic. All 69 CAS procedures were technically successful. The major post operative complications were two minor strokes (2.8%), one major stroke (1.4%) and one fatal major stroke (1.4%). The mean length of follow-up was 17 months (range, 4 months to 5.6 years). Two patients (2.8%) have suffered ipsilateral neurologic events following CAS. Long-term follow-up revealed restenosis at 6 months in 4 patients (5.7%). CONCLUSIONS: Carotid artery angioplasty and stenting is safe and feasible. This procedure produces satisfactory outcomes in patients who are at high risk for post-operative complications of CEA. PMID- 11385153 TI - Carotid artery stenting: careful case selection and meticulous technique produce excellent results. PMID- 11385154 TI - AAA stent grafts: current developments. PMID- 11385155 TI - Late stent thrombosis after successful rescue of a major coronary artery rupture with a polytetrafluoroethylene-covered stent. AB - We describe a case in which we successfully treated a major left anterior descending artery rupture with a polytetrafluoroethylene-coated stent. The patient presented with acute antero-apical myocardial infarction 52 days after the initial procedure and cardiac catheterization revealed late stent thrombosis, which was successfully treated by primary angioplasty. PMID- 11385156 TI - Primary angioplasty and stenting in a patient with primary antiphospholipid syndrome and thrombocytopenia. AB - Antiphospholipid syndrome is characterized by venous and arterial thrombosis, thrombocytopenia, stroke and, rarely, acute coronary syndromes. However, there are no data available regarding the management of acute myocardial infarction in primary antiphospholipid syndrome with accompanying severe thrombocytopenia and cardiogenic shock. We describe such a case, which was managed by successful primary percutaneous transluminal coronary angioplasty and stent implantation with accompanying immunosuppression therapy. PMID- 11385157 TI - "Wireless" laser recanalization of chronic total coronary occlusions. AB - Chronic total occlusions in particular, completely obstructed aorto-ostial lesions are among the most challenging targets in interventional cardiology. Excimer laser is a debulking technology for revascularization of complex lesions. Treatment of total occlusions with laser angioplasty can be applied providing that a guidewire traverses the entire length of the occlusion prior to device activation. In many patients with total occlusions, a guidewire is unable to penetrate the target stenosis. This communication presents a new technique termed "wireless" laser recanalization. This approach entails recanalization of a total occlusion with a laser catheter without a leading guidewire. PMID- 11385158 TI - Inappropriate ICD therapy due to lead-related noise in an active fixation ICD lead. AB - INTRODUCTION: Despite advances in technology, problems with effective ventricular endocardial sensing continue to arise and to result in inappropriate implantable cardioverter defibrillator (ICD) therapies. METHODS AND RESULTS: The patient is a 79-year-old man with ischemic cardiomyopathy and ventricular tachycardia (VT) who received inappropriate ICD therapies due to lead-related noise detection. Noise was created by the distal helix, which was not fully deployed, intermittently contacting the internal guidepost of the lead (CPI, Guidant, model 0155). After the complete deployment of the helix the problem resolved and the pacing parameters remained stable. CONCLUSION: To our knowledge, this is the first confirmed case report of non-physiologic sensing secondary to the incompletely deployed helix of an active fixation defibrillation lead. A design flaw with this lead led to its redesign to avoid noise over-sensing. PMID- 11385160 TI - Coronary surgery via left mini-thoracotomy (West coast technique). PMID- 11385159 TI - Vascular brachytherapy: what have we learned from intravascular ultrasound? AB - With the advent of intravascular brachytherapy (IVBT), a striking reduction in the rate of restenosis has been observed. The use of intravascular ultrasound (IVUS) during IVBT trials has shown many aspects and relevant pathophysiologic mechanisms following this practical therapy. Specifically, IVUS quantitative assessments have demonstrated a drastic inhibition of both neointimal formation and negative remodeling to be the predominant vascular response to IVBT. Moreover, IVBT has shown promise for challenging high-risk restenosis cases, such as patients with diabetes mellitus and smaller caliber target vessels. However, unexpected radiation-induced complications have also been discovered, as more patients have been treated for a broad class of lesion subsets. Edge effect, induced by catheter-based radiation, was seen to be due to geographic miss of the radiation source, advocating a new concept known as "radiation edge". Furthermore, late thrombosis, which is known to be strongly associated with new stent implantation following IVBT, may be avoided with novel antiplatelet agents. Two additional complications, whose clinical significance remains unclear, are unhealed dissection and late stent malapposition featured by IVUS qualitative assessment. Unhealed dissection was observed in half of radiated dissections and late stent malapposition has been seen for all radiation sources in a small percentage of cases at 6-12 months follow-up. Radiation sources, dosimetry, and delivery methods continue to improve and should ultimately translate to more effective treatment for the patient with atherosclerotic coronary disease. PMID- 11385161 TI - Is cardiac surgery here to stay in coronary artery disease? PMID- 11385162 TI - Digital imaging and archiving in a radiological department: the 2nd Radiology Department's Experience at the University of Athens. PMID- 11385163 TI - Efficacy and safety of ticlopidine monotherapy versus ticlopidine and aspirin after coronary artery stenting: follow-up results of a randomized study. AB - A combined antiplatelet treatment with ticlopidine and aspirin has been accepted as standard pharmacological regimen after coronary artery stenting. No data of a randomized trial are available on ticlopidine monotherapy. This prospective, randomized monocenter trial investigates the role of ticlopidine monotherapy versus combined antiplatelet therapy with ticlopidine and aspirin in unselected patients undergoing coronary artery stenting. After successful placement of 378 coronary artery stents, two hundred and forty-three consecutive patients were randomly assigned to receive antiplatelet therapy with 2 x 250 mg ticlopidine (121 patients) or a combination of 2 x 250 mg ticlopidine plus 100 mg aspirin (122 patients) daily. The primary endpoint included the absence of death, cardiac events and vascular access-site complications during the in-hospital phase. Angiographic and clinical assessment was repeated at the 3-month follow-up exam. Two hundred and thirty-seven patients (97.5%) were free from cardiac and non cardiac events. Stent thromboses were seen in 2 patients of the combined treatment group, while none were observed in the monotherapy group. No statistically significant differences were found between the 2 groups regarding the primary endpoint. Angiography performed in 210 patients (86.4%) at follow-up revealed a restenosis rate of 29.4% in the combined treatment group and 27.8% in the monotherapy group. Monotherapy with ticlopidine is as safe and effective as a combined regimen of ticlopidine plus aspirin after coronary artery stenting in an unselected patient population. These results need to be confirmed in a larger multicenter trial. PMID- 11385164 TI - Ticlopidine monotherapy following coronary stent deployment: "penny wise and pound foolish". PMID- 11385165 TI - Efficacy of cutting balloon angioplasty for in-stent restenosis: an intravascular ultrasound evaluation. AB - BACKGROUND: The increase in the use of stents has seen the increasing emergence of in-stent restenosis (ISR). Reports suggest that the Cutting Balloon (Interventional Technologies, San Diego, California) may be a useful treatment modality for this new clinical entity. METHODS: In this study, we compared the efficacy of Cutting Balloon angioplasty (CBA) with conventional balloon angioplasty (PTCA) for ISR in 47 patients (47 lesions). Results were evaluated with intravascular ultrasound (IVUS). The CBA group included 25 patients (mean age, 65 +/- 78 years; 7 females) and the PTCA group included 22 patients (mean age, 69 +/- 51 years; 6 females). RESULTS: The procedural success rate was 100% in both groups. IVUS showed that luminal area acute gain was larger in the CBA group (2.5 +/- 0.8 mm2) compared to the PTCA group (1.8 +/- 1.0 mm2), while late loss was smaller in the CBA group (0.5 +/- 0.4 mm2) compared to the PTCA group (1.3 +/- 0.5 mm2). The change in total area was similar in both groups. The increase in area at the stented portion was 0.4 +/- 0.8 mm2 in the CBA group and 1.2 +/- 0.5 mm2 in the PTCA group. The restenosis rate at follow-up (mean follow up, 5.4 months) was higher in PTCA patients (59%) than in the CBA patients (24%). CONCLUSION: CBA may result in no increase in total vessel area, a constant stent area, a decrease in plaque area, and an increase in lumen area (induced by the decrease in plaque area). Our IVUS findings suggest that compared to PTCA, the dilatation mechanism of CBA may be associated with reduced dilatation of both the total vessel area and the stent area for ISR. The mechanism of this modality may minimize injury to the intimal membrane and may potentially be a primary device for in-stent restenosis in the future. PMID- 11385166 TI - The cutting balloon: a significant advance in the treatment of coronary artery disease. PMID- 11385167 TI - Percutaneous intervention in saphenous venous grafts: in-stent restenosis lesions are safer than de novo lesions. AB - BACKGROUND: The histological appearance of stenosis in de novo saphenous venous grafts (DNSVG) consists of diffuse atherosclerosis that contains blood elements, necrotic debris and limited fibrocollagenous tissue. The friable nature of these lesions complicates percutaneous intervention (PCI) procedures. On the other hand, in-stent restenosis (ISR) of SVG is due primarily to atherosclerotic plaque or fibromuscular hyperplasia, with thrombus formation playing a secondary role. The purpose of this study is to compare the results of PCI in these two types of SVG lesions. METHODS: We reviewed our institutional interventional database from March 1996 through February 2000 and identified all consecutive patients who underwent PCI of at least one SVG. One hundred and ten patients were identified: 89 undergoing DNSVG intervention and 21 patients with ISR lesions. RESULTS: Acute coronary syndromes, degenerated and thrombus-containing lesions were more common in the DNSVG group. "Slow-, no-reflow" complicated 20% of the DNSVG lesions compared to none of the ISR lesions (p = 0.02). Post-procedural myocardial infarction was higher in the DNSVG group (13.5% versus 0%; p = 0.1) and correlated significantly with the occurrence of "slow-, no-reflow" (r = 0.43; p = 0.0001). Utilizing statistical modeling to adjust for baseline differences between the groups, ISR lesions were associated with a low risk of procedural complications (r = 0.22; p = 0.03). CONCLUSION: This study demonstrates that in this relatively high-risk population, PCI is safer in ISR lesions than in de novo SVG lesions. PMID- 11385168 TI - Distal embolization and no-reflow in the setting of saphenous vein graft in-stent restenosis. PMID- 11385169 TI - Saphenous vein graft interventions: still far away from the goal. PMID- 11385170 TI - A new device for percutaneous suture-mediated closure of arterial puncture sites using exteriorized needles: initial experience. AB - PURPOSE: To assess the feasibility and initial clinical results of Closer (Perclose, Redwood City, California), a new device for percutaneous suture mediated closure of arterial puncture sites. METHODS: Vascular interventions were performed using 6 and 7 French sheaths in 22 consecutive patients. All patients underwent suture-mediated percutaneous closure with the new device. Patients were followed by physical examination the day after the procedure. RESULTS: Immediate hemostasis was achieved in 20/22 patients (91%). In 3 out of 5 antegrade closures, only one suture was captured. In two cases, this was managed by reinsertion of a second device. In another patient, hemostasis was not achieved by the device due to knot entrapment. No major complications were observed. The overall rate of minor complications was 9% (2/22) due to hematomas without the need for blood transfusions. CONCLUSION: Percutaneous suture with this new device is an acceptably safe and effective method to achieve immediate hemostasis of the arterial access site after interventional procedures with minimized manipulation of the puncture track. PMID- 11385171 TI - Rota-stenting of a calcified anomalous left coronary artery via the arm in a multi-vasculopathic patient. AB - Anomalous origins of the coronary arteries pose a challenge to the interventionalist. Diagnostic testing must take into account the possibility that perfusion patterns may be unusual but not pathologic, resulting in false-positive perfusion scanning. Treatment strategies must be customized to allow unusual access and passage of devices around angulations not commonly encountered. In this report, we describe a patient with severe peripheral arterial disease requiring the use of a 7 French multipurpose guide via the brachial approach for rotational atherectomy and stenting of a calcified and angulated anomalous left coronary. PMID- 11385172 TI - Adventitial hematoma triggering coronary spasm during percutaneous coronary intervention. AB - During percutaneous coronary intervention of the left anterior descending coronary artery, a lumen narrowing was observed proximal to the stent just deployed. Intravascular ultrasound showed a hematoma localized outside the trilaminar wall structure in absence of a dissection flap or evidence of compression of the lumen. The luminal narrowing resolved after intracoronary administration of vasodilators. This finding is compatible with a coronary spasm triggered by an adventitial hematoma following stent deployment. PMID- 11385173 TI - Spontaneous left anterior descending artery to right ventricular fistula. AB - We report a very unusual case of a spontaneously acquired left anterior descending to right ventricular fistula. We believe that this spontaneous occurrence was the result of hemodynamic changes that occurred between the first and second catheterizations. PMID- 11385174 TI - Successful use of eptifibatide as an adjunct to coronary stenting in a patient with abciximab-associated acute profound thrombocytopenia. AB - A 72-year-old male who was given abciximab for unstable angina developed acute profound thrombocytopenia with a platelet count nadir of 6,000/mm3. He was treated with steroids and platelet transfusion. Four days later, he underwent coronary angioplasty after pretreatment with eptifibatide without development of thrombocytopenia. This suggests that the development of thrombocytopenia with abciximab is not necessarily a contraindication to subsequent use of glycoprotein (GP) IIb/IIIa receptor antagonists. Eptifibatide may be an appropriate consideration in high-risk patients who would benefit from a GP IIb/IIIa receptor antagonist, in spite of acute profound thrombocytopenia due to abciximab therapy. PMID- 11385175 TI - Stent implantation in small coronary vessels. PMID- 11385176 TI - Sealing versus suturing devices. PMID- 11385177 TI - Percutaneous coronary intervention and subsequent endovascular beta-radiation brachytherapy in a 14-year-old with repaired anomalous left coronary artery from pulmonary artery. AB - Anomalous origin of the left coronary artery from pulmonary artery (ALCAPA) is a rare congenital anomaly. Re-establishment of the dual coronary system is the standard treatment, although the long-term outcome after surgical repair is not well defined. We report a case in which coronary stenting was performed to treat left anterior descending artery lesions eight months after surgical repair of ALCAPA. The patient then developed rapid in-stent restenosis within three months, which was successfully treated by rotational atherectomy, balloon angioplasty, and catheter-based beta-radiation brachytherapy. Follow-up angiograms after three and six months showed no recurrent in-stent restenosis. This represents the first report of coronary stenting in the setting of ALCAPA, and the first report of catheter-based intracoronary radiation therapy in a pediatric patient. PMID- 11385178 TI - Project on telemedicine, and the unique professional atmosphere of the Mayo Clinic and the Interventional Andreas Gruentzig Society: an interview with Kirk N. Garratt, MD. PMID- 11385179 TI - The disintegrin kistrin inhibits neurite extension from spiral ganglion explants cultured on laminin. AB - The influence of laminin-1 (LN) and tenascin-C (TN), extracellular matrix molecules expressed spatially and temporally along the neural growth route from spiral ganglion (SG) neurons to the cochlear sensory cells, was evaluated in cultured SG explants from postnatal day 4 rats. Increasing concentrations of LN resulted in a strong, dose-dependent increase in the length of neurites and in a higher number of neural processes, while varying TN concentrations had relatively minor effects on both parameters. The results suggest differential receptor activation by LN and TN. When explants grown on LN were treated with Kistrin, an inhibitor of the alphavbeta3 integrin, the LN-induced increase in neurite length was reduced in a dose-dependent manner. However, the number of extending neurites was not affected, indicating that different receptors mediate this response, perhaps by increasing neuronal survival. PMID- 11385180 TI - Functional recovery of hearing following ampa-induced reversible disruption of hair cell afferent synapses in the avian inner ear. AB - Hair cells in the avian inner ear can regenerate after acoustic trauma or ototoxic insult, and significant functional recovery from hearing loss occurs. However, small residual deficits remain, possibly as a result of incomplete reestablishment of the hair cell neural synaptic contacts. The aim of the present study was to determine if intracochlear application of alpha-amino-3-hydroxy-5 methyl-4-isoxazole propionic acid (AMPA), an excitotoxic glutamate agonist, causes reversible disruption of hair cell neural contacts in the bird, and to what extent functional recovery occurs if synaptic contacts are reestablished. Compound action potential (CAP) responses to tone bursts were recorded to determine hearing thresholds during a recovery period of up to 4 months. Subsequently, the response properties of single auditory nerve fibers were analyzed in the same animals. Instillation of AMPA into the perilymph of the scala tympani led to immediate abolition of CAP thresholds. Partial recovery occurred over a period of 2-3 weeks, without further improvement of thresholds thereafter. High-frequency thresholds did not reach control values even after 3-4 months of recovery. Single-ganglion cell response properties, obtained 3-4 months after AMPA treatment, showed elevated thresholds at the fiber's characteristic frequency (CF) for units with CF above 0.3 kHz. Sharpness of tuning (Q(10 dB)) was reduced in units with CF above 0.4 kHz. The spontaneous firing rate was higher in units with CF above 0.18 kHz. The maximum sound-evoked discharge rate was also increased. Transmission electron micrographs of the basilar papilla showed that, following AMPA treatment, the nerve endings went through a sequence of swelling, degeneration and recovery over a period of 3-7 days. The process of neosynaptogenesis was completed 14 days after exposure. The present findings are strong evidence for a role of glutamate or a related excitatory amino acid as the afferent transmitter in the avian inner ear. In addition they show that functional recovery after disruption and regeneration of hair cell neural synapses, without apparent damage to the hair cells, is incomplete. PMID- 11385182 TI - Contralateral masking in cochlear implant users with residual hearing in the non implanted ear. AB - Contralateral masking was investigated in cochlear implant users with residual hearing in the non-implanted ear. Threshold elevations for acoustic probes were observed when electrical maskers were presented in the opposite ear. Also, threshold elevations for electrical probes were observed when acoustic contralateral maskers were presented. The amount of threshold shift expressed in decibels charge or decibels sound pressure level produced by either contralateral acoustic or electric maskers was within the range found in normal listeners for similar stimuli (i.e. 4-8 dB). There was a correlation between the sensation level of acoustic maskers and the maximum amount of masking observed which is consistent with data for normally hearing subjects. The width of the masking patterns was similar to that expected from forward masking patterns in severely sensorineurally impaired ears and implanted ears. The maximum amount of acoustic masking tended to occur for electrode positions that were more basal than expected from characteristic frequency positions. However, where a relatively high-frequency 4-kHz masker could be used, there was a good match between the characteristic frequency position of the maximum threshold elevation and that of the masker. PMID- 11385181 TI - Use of the pinna reflex as a test of hearing in mutant mice. AB - Although it is a gross measure, the pinna reflex test is easily administered and is, therefore, incorporated as a general screening tool in mutagenesis programs. Our recent application of this approach indicated that mutant mice lacking one of the small Maf proteins, in this case MafG, failed to exhibit a pinna reflex. In contrast, littermate controls, with the same mixed 129/CD1 background, and including both wild type and heterozygous mutant animals, passed the test. Because previous studies indicate that mafG is expressed in both cochlear and vestibular parts of the mouse inner ear, the source of this 'presumed deafness' was further assessed by making round window recordings to determine compound action potential thresholds. Auditory brainstem responses were also acquired to assess function along portions of the central auditory pathway. In all cases, responses in homozygous mutants (-/-) were comparable to those obtained from littermate controls, either wild type (+/+) or heterozygous mutants (+/-). Gross anatomy of the organ of Corti was also found to be similar in all three groups of mice. Hence, the lack of a pinna reflex must relate to nonauditory causes. PMID- 11385183 TI - Directional sensitivity of the human macula utriculi based on morphological characteristics. AB - A morphometric analysis of 43 human maculae utriculi is presented. Individual data on the shape, total area and relative area of the pars interna are given. In addition, the sensitivity of the entire macula to shear stimuli in different directions was estimated. The mean area of 39 maculae from adults and children was 4.30 +/- 0.30 (SD) mm(2). The pars interna was slightly but significantly smaller than the pars externa. The interindividual variability was larger for the shape of the macula than for the total area and the percentage of the pars interna. The estimated responsiveness of the macula was largest for shear directed anteromedially and smallest for shear directed posteriorly. The data are discussed taking into consideration clinical findings on patients with unilateral loss of otolith function. PMID- 11385184 TI - Antiarrhythmic agents in facilitating electrical cardioversion of atrial fibrillation and promoting maintenance of sinus rhythm. AB - Atrial fibrillation (AF) is a prevalent arrhythmia associated with significant morbidity and mortality. Electrical cardioversion of AF is a potentially definitive treatment, but as little as 67% of patients may be successfully cardioverted and, after normal sinus rhythm (NSR) is achieved, AF often recurs. Class IA, IC, and III antiarrhytmic agents are used for both facilitation of electrical cardioversion and subsequent maintenance of NSR. The mechanisms of these agents may be related to suppressing automaticity, prolonging the wavelength of reentrant wavelets, and preventing electrical remodeling. The possibility of proarrhythmia and other adverse effects complicates use of these drugs, and no large trials have been completed to elucidate definite indications. Several factors may predict failure with electrical cardioversion alone (duration of AF, atrial size, age, underlying disease, and factors that affect transthoracic impedance), calling for empiric pharmacotherapy to facilitate cardioversion. For this purpose, class IA agents hold some promise, evidence for class IC agents is conflicting, and class III agents are the most effective. Adverse effects are rare given the short course before cardioversion, but ibutilide, the most efficacious in this regard, may be proarrhythmic after only a single dose. In promoting maintenance of sinus rhythm, antiarrhythmics across the different classes have similar efficacies: NSR may be maintained in approximately 40-65% of patients compared to approximately 30-35% with placebo at 1 year. Amiodarone is distinct in its success, with approximately 60-80% of patients remaining in NSR. For all of these agents, long-term therapy may lead to proarrhythmia or other substantial adverse effects. Finally, a serial antiarrhythmic strategy may be effective, with maintenance of NSR and minimal adverse effects ultimately achieved by trial and error. PMID- 11385185 TI - HMG-CoA reductase inhibitors: Is the endothelium the main target? AB - Endothelial dysfunction is an early event in atherosclerosis and could be considered a response to the injury induced by major risk factors. There is evidence that endothelial dysfunction is intimately involved in the onset and the progression of cardiovascular disease through abnormalities in the production, release or degradation of endothelium-derived factors, mainly nitric oxide and endothelin 1. Several reports have shown that drugs of the statin class could have multiple beneficial effects related to endothelium-mediated vasoactive, antithrombotic, antiproliferative and anti-inflammatory actions. Thus, the question arises of whether endothelial cells are the main target of statin therapy, in the setting of both hypercholesterolemia and normocholesterolemia. Experimental and clinical studies are reported that could support this hypothesis. PMID- 11385186 TI - Helicobacter pylori infection and coronary heart disease in Japanese patients. AB - Although several independent studies have claimed a link between Helicobacter pylori infection and coronary heart disease (CHD), this association has not been established conclusively. The aim was to determine whether an association between H. pylori infection and CHD can be demonstrated in Japanese patients. Three hundred and four patients who underwent consecutive coronary arteriography were investigated. Ninety-four patients had single-vessel coronary stenosis and 112 had multi-vessel stenosis. The remaining 98 patients had no significant stenosis in any coronary arteries. H. pylori infection was diagnosed serologically and the association between infection and CHD was estimated by the odds ratio. The serum pepsinogen (PG) I-II ratio was used to estimate the degree of gastric atrophy. Seropositivity for H. pylori was significantly higher in the patients with CHD (67%) than in the controls (50%; p = 0.006). The odds ratio for CHD after having H. pylori infection was estimated as 1.35 (95% confidence interval 1.03-1.78; p = 0.028), after adjustment for the common risk factors of CHD in a logistic regression analysis. The association between CHD and H. pylori infection was more significant among patients without any history of diabetes or smoking. The PG I II ratio in H. pylori-positive patients was significantly higher in the multi vessel group (3.46) than in the control or single-vessel group (2.86, p = 0.030; 2.78, p = 0.008; respectively). H. pylori infection was shown to be an independent risk factor for CHD in Japanese patients, especially among those who did not have a history of diabetes or smoking. These data imply that the association between H. pylori infection and CHD is clinically relevant. PMID- 11385187 TI - Coronary artery disease but not coronary calcification is associated with elevated levels of cardiolipin, beta-2-glycoprotein-I, and oxidized LDL antibodies. AB - BACKGROUND: Autoimmune factors have been shown to play a role in atherosclerosis. The aim of this study is to correlate 5 autoantibodies (anticardiolipin, anti-CL, beta2-glycoprotein-I, beta2GPI, phosphatidylcholine, oxidized low-density lipoprotein, oxLDL, endothelial cell) with the presence of coronary heart disease, angiographic findings, and with coronary artery calcification. METHODS: The levels of the 5 autoantibodies and a control antifibroblast line of 126 coronary heart disease patients and 20 healthy controls were measured. Fifty-one patients underwent coronary angiography, and 98 patients had coronary artery calcium determination using spiral computerized tomography (dual mode). RESULTS: Levels of 3 autoantibodies (anti-CL, beta2GPI, oxLDL) were significantly elevated in coronary heart disease patients compared with controls (p < 0.001, p = 0.001, p < 0.001, respectively). Within the subgroup of patients with significant coronary artery stenosis, anti-CL antibodies were also elevated (p = 0.008). No correlation was found between anti-CL, and anti-beta2GPI autoantibody levels and coronary calcium scores as measured by spiral computerized tomography. However, anti-oxLDL antibodies were raised in patients with no calcification detected by spiral computerized tomography, compared with the patients with any coronary calcification (p = 0.046). CONCLUSION: Anti-CL, beta2GPI and oxLDL antibodies are elevated in coronary heart disease patients regardless of coronary calcification. PMID- 11385188 TI - Prevalence of heart disease in asymptomatic chronic cocaine users. AB - To determine the prevalence of heart disease in outpatient young asymptomatic chronic cocaine users, 35 cocaine users and 32 age-matched controls underwent resting and exercise electrocardiography (ECG) and Doppler echocardiography. Findings consistent with coronary artery disease were detected in 12 (34%) patients and 3 (9%) controls (p = 0.01). Decreased left ventricular systolic function was demonstrated in 5 (14%) patients, but in none of the controls (p = 0.055). Finally, resting and peak exercise abnormal left ventricular filling was detected in 38 and 35% of patients as compared to 19 and 9% of controls, respectively (p = 0.11 and 0.02, respectively). We conclude that coronary artery or myocardial disease is common (38%) in young asymptomatic chronic cocaine users. Therefore, screening ECG and echocardiography may be warranted in these patients. PMID- 11385189 TI - Insulin resistance is increased by transdermal estrogen therapy in postmenopausal women with cardiac syndrome X. AB - Estrogen has been reported to have both short- and long-term effects on the cardiovascular system. However, it remains to be examined how short-term transdermal estrogen therapy (TET) affects insulin sensitivity (SI) in patients with cardiac syndrome X (CSX), who are characterized by elevated insulin resistance. SI was assessed in a randomized, double-blind, placebo-controlled crossover study by minimal model analysis in seven postmenopausal women with CSX treated by TET. SI decreased by 32 +/- 8.3%, from 5.94 +/- 1.14 at baseline to 3.61 +/- 0.40 [(10(-4) x min(-1))/(microU/ml)] during TET (p = 0.03). Time to the onset of symptoms increased from 414.2 +/- 51.0 s at baseline to 450.0 +/- 53.2 s (p = 0.04). We conclude that TET increases SI in postmenopausal women with CSX. This effect is unrelated to the beneficial anti-ischemic effects on exercise duration. PMID- 11385190 TI - The expression of the CD18 leucocyte integrin in a rabbit model of acute myocardial infarction: a pilot study of temporal changes and relationship to infarct size. AB - CD18 integrins mediate leucocyte adhesion to vascular endothelium. This represents the initial step in inflammatory cell infiltration following myocardial necrosis. The current study assessed whether the expression of these rapidly activated and readily measured adhesion receptors on circulating neutrophils would reflect the extent of cardiac damage in a rabbit model of acute myocardial infarction. Myocardial ischaemia/infarction was induced in anaesthetised adult male New Zealand white rabbits (n = 8) by ligation of the circumflex or marginal coronary artery. To control for the effects of anaesthesia and surgery, 4 rabbits underwent identical procedures without the induction of infarction. Absolute infarct size (in mg) and infarct size as a percentage of total left ventricular mass (relative infarct size) were calculated by differential staining and weighing of necrotic myocardium. Flow cytometry was used to determine cell surface expression of CD18 at six time points (baseline, 20 and 45 min of ischaemia and 20, 60 and 180 min of reperfusion). Absolute neutrophil CD18 expression and changes in expression over baseline were correlated with absolute and relative infarct size. Mean neutrophil CD18 expression increased significantly (from 2.42 +/- 0.20 to 3.07 +/- 0.29; p = 0.04) within 20 min of ischaemia. CD18 expression at 3 h did not predict absolute or relative infarct size (r = 0.40 and 0.37, respectively). The percentage change in cell surface CD18 expression (above baseline levels) was, however, correlated with both measures of infarct size (r = 0.76, p = 0.03, and r = 0.92, p = 0.001, respectively). In conclusion, in this rabbit model of myocardial infarction, neutrophil CD18 expression rises within 20 min of the induction of ischaemia but absolute values after 3 h of reperfusion are poor indicators of infarct size. Although percentage change in neutrophil CD18 levels over baseline correlates with infarct size there is considerable variation between individuals, limiting any clinical application. PMID- 11385191 TI - In vivo magnetic resonance imaging and surgical histopathology of intracardiac masses: distinct features of subacute thrombi. AB - We evaluated intracardiac masses in vivo, in situ and histologically to determine tissue properties revealed by magnetic resonance (MR) imaging. In 15 consecutive patients scheduled for cardiotomy, the cardiac chambers were studied preoperatively with MR imaging and echocardiography. Visual examination of one or more chambers was performed during cardiotomy for mitral valve replacement, aneurysmectomy, atrial septal repair and atriotomy. Six thrombi (1 atrial appendage, 5 ventricular) and 2 atrial myxomas were removed and subjected to histological analysis. All masses were detected preoperatively by MR imaging. The smallest was a subacute 3-mm mural clot in the left ventricle and was undetected by transesophageal and transthoracic echocardiography. The 3 subacute clots had homogeneously low MR signals, did not enhance with gadolinium and exhibited magnetic susceptibility effects; histopathology confirmed these clots to be avascular and laden with dense iron deposition related to hemoglobin breakdown products. The 3 organized clots had intermediate and heterogeneous MR signals and multiple areas of gadolinium enhancement. The 2 myxomas had low MR signals and gadolinium enhancement in the core and septal attachment; these areas had dense neovascular channels. Subacute thrombi appear to have MR features that are distinct from organized thrombi and myxomas, and MR images of subacute thrombi contrast sharply with normal cardiac structures, enabling detection of thin mural clots that may be echographically occult. These findings may be of value, because a subacute clot may be more likely than an organized thrombus to give rise to an embolus. PMID- 11385192 TI - Severe intracranial bleeding mimicking acute inferior myocardial infarction with right ventricular involvement. AB - Electrocardiographic (ECG) changes and wall motion abnormalities of the left ventricle have been observed in patients with severe intracranial hemorrhage. However, ECG evidence of an acute myocardial infarction in this setting is extremely rare but may have important therapeutic consequences. We report the case of a 45-year-old female who became unconscious with respiratory insufficiency after an endoscopic retrograde cholangiopancreaticoscopy with ECG changes consistent of an inferior myocardial infarction with right ventricular involvement. Immediate coronary angiography revealed normal coronaries; however, left ventricular angiography showed extensive wall motion abnormalities predominantly in the anteroseptal region. Immediate cranial computer tomography demonstrated massive intracranial bleeding. Intracranial hemorrhage can be associated in the initial phase with ECG evidence of an acute myocardial infarction. This has to be taken into consideration in the setting of unexplained loss of consciousness or nonresponsiveness of a patient. A rapid diagnostic evaluation has to be initiated to rule out a myocardial infarction and to diagnose intracranial hemorrhage before the use of thrombolytic or anticoagulant therapy. PMID- 11385193 TI - Defining the meaning of exercise-induced S-T segment elevation in Q wave leads in postinfarction patients using a new ECG parameter of transmural ischemia: the stress-induced Q-Tc interval shortening. PMID- 11385194 TI - Dental amalgam fillings and the amount of organic mercury in human saliva. AB - We studied differences in the amounts of organic and inorganic mercury in saliva samples between amalgam and nonamalgam human study groups. The amount of organic and inorganic mercury in whole saliva was measured in 187 adult study subjects. The mercury contents were determined by cold-vapor atomic absorption spectrometry. The amount of organic and inorganic mercury in paraffin-stimulated saliva was significantly higher (p<0.001) in subjects with dental amalgam fillings (n = 88) compared to the nonamalgam study groups (n = 43 and n = 56): log(e) (organic mercury) was linearly related to log(e) (inorganic mercury, r(2) = 0.52). Spearman correlation coefficients of inorganic and organic mercury concentrations with the number of amalgam-filled tooth surfaces were 0.46 and 0.27, respectively. Our results are compatible with the hypothesis that amalgam fillings may be a continuous source of organic mercury, which is more toxic than inorganic mercury, and almost completely absorbed by the human intestine. PMID- 11385195 TI - Fluoride concentrations of unstimulated whole and labial gland saliva in young adults after fluoride intake with milk. AB - Following a fluoride depletion period 6 subjects repeatedly rinsed with 30-ml volumes of milk with or without added fluoride (5 ppm). Rinsing time was timed according to the measurements on how long it took to drink 200 or 500 ml milk. Rinsing with fluoridated milk for a total interval of 20 and 60 s, respectively, did not influence significantly the fluoride concentration of unstimulated centrifuged whole saliva 45 min later. Neither the urinary fluoride concentration nor the fluoride excretion were significantly affected by rinsing with fluoridated milk during the first hour. However, intake of 1 or 2.5 mg of fluoride with 200 and 500 ml milk, respectively, resulted in significant elevations in whole saliva fluoride levels 45 min later. In addition, the fluoride excretions into urine produced during 60 min after the fluoride intakes were significantly elevated and those reflected the ingested dose of fluoride. The intake of either 1 or 2.5mg fluoride with milk did not significantly influence the fluoride level of unstimulated labial gland saliva collected simultaneously with whole saliva. PMID- 11385196 TI - Influence of maternal xylitol consumption on mother-child transmission of mutans streptococci: 6-year follow-up. AB - Xylitol is effective as a noncariogenic or even cariostatic sugar substitute. Habitual xylitol consumption appears to select for mutans streptococci (MS) which shed easily into saliva from plaque. We have earlier shown that habitual xylitol consumption of mothers was associated with a statistically significant reduction in the probability of mother-child transmission of MS assessed at 2 years of age. The aim of the present study was to assess the children's MS counts 1 and 4 years after the maternal xylitol consumption had been discontinued. At baseline, during pregnancy, all mothers (n = 195) showed high salivary levels of MS. The mothers were randomly assigned to xylitol, fluoride (F) and chlorhexidine (CHX) groups. In the xylitol group, the mothers chewed xylitol-sweetened gum, for 21 months, starting 3 months after delivery. In the two control groups, the mothers received CHX or F varnish treatments at 6, 12 and 18 months after delivery. At the 2-year examination, 169 mother-child pairs participated. At the 3-year and 6-year examinations, there were 159 and 147 children in the study, respectively. For children's MS analyses, visible plaque was collected using toothpicks at the age of 3 and paraffin-stimulated saliva at the age of 6. The persons involved in the collection and analysis of the microbiological samples were blinded as to the study design and group. Both the plaque and salivary MS were cultured on Mitis salivarius agars containing bacitracin. In all groups, the colonization percentages increased during the follow-up. At the 3-year examination, the children's risk of having MS colonization was 2.3-fold in the F group (95% CI 1.3 4.2) compared to the xylitol group. This difference was statistically significant. Even at 6 years of age, the salivary MS levels were significantly lower in the xylitol group than in the other groups (ANOVA, p<0.001). In conclusion, the earlier demonstrated, xylitol-associated reduction in the probability of mother-child transmission of MS was still found in the children's MS counts at the age of 3 and 6 years. PMID- 11385197 TI - Influence of approximal caries in primary molars on caries rate for the mesial surface of the first permanent molar in swedish children from 6 to 12 years of age. AB - The objective was to assess the influence on the caries rate for the mesial surface of the first permanent molar (6m) of the caries status of the distal surface of the second primary molar (05d) in children from 6 to 12 years of age. The study design was retrospective and included 374 children with an average of 5 sets of bite-wing radiographs. The mean age of the children was 6.7 years when the first bite-wing radiographs were taken and 11.5 years at the time of the latest radiographs included in the study. The approximal surfaces were classified according to a scoring system: 0 = no visible radiolucency, 1= radiolucency in the outer half of the enamel, 2 = radiolucency in the inner half up to the enamel dentin border, 3 = radiolucency with a broken enamel-dentin border but with no obvious involvement of the dentin, 4 = radiolucency with obvious spread in the outer half of the dentin, and score 5 = radiolucency in the inner half of the dentin. The influence of the status of 05d on the caries rate for 6m (state > or =2) was assessed by using a model for dependence between the two neighbouring surfaces. Presence or absence of approximal caries in the distal surface of the first primary molars (04d) and/or the mesial surface of the second primary molars (05m) at the time of eruption of 6m was also related to the caries rate for 6m. The caries rate for 6m was 15 times higher if 05d had developed enamel/enamel dentin border caries (state 2 or 3) compared to a radiographically sound 05d (state 0 or 1). No significant increase in the caries rate for 6m was found when 05d had deeper unrestored dentin caries than when 05d had superficial caries (state 2 or 3). Furthermore, for preventing caries in 6m, no benefit could be found from restoring an 05d with dentin caries with amalgam as opposed to leaving the carious lesion unrestored until exfoliation. The caries rates for 6m and 05d were 3.4 and 2.7 times higher, respectively, in the presence of dentin caries/restoration in 04d/05m compared to absence of dentin caries in these molar surfaces at the time of eruption of 6m. In conclusion, the caries rate for 6m depended on the status of 05d and increased 15 times if 05d had enamel/enamel dentin border caries compared to a sound 05d. Deeper unrestored or restored dentin lesions in 05d did not result in a higher caries rate for 6m compared with more shallow carious lesions in 05d. PMID- 11385198 TI - Prognosis of caries increment with past caries experience variables. AB - This study aimed to select past caries experience variables as caries predictors and to compare their prognostic accuracy with the variables used in a method of caries prognosis developed in a Swiss child population. The data used for the analyses originated from an 8-year longitudinal study starting in 1980 on caries of 7.5-year-old Dutch children. Stepwise logistic regression analyses provided predictor variables. The newly introduced variables D23fi, D(123)i and D23pifi were interchangeable and the most powerful caries predictors. For the sake of uniformity and ease of application, D23fi (number of fissures of the permanent first molar with non-cavitated or cavitated caries lesions) was chosen as the first variable in the logistic regression equations. The gain in accuracy of the second and third predictor variables (number of sound primary molars and the number of buccal and lingual smooth surfaces of the permanent first molar with non-cavitated or cavitated caries) in the regression equations was limited. The D1 condition of surfaces could be omitted from the prediction models. The present forced three-predictor-regression equations for 7.5-, 9.5- and 11.5-year-old children were evaluated to assess their prognostic performance by using the area under the ROC curve as a measure of prognostic quality. For the present regression equations, the area under the ROC curve was 81-87%, which was higher compared to the Swiss regression equations for caries prognosis. PMID- 11385199 TI - Conventional, digital and contrast-enhanced bitewing radiographs in the decision to restore approximal carious lesions. AB - The purpose of the present investigation was to compare the accuracy of treatment decisions in proximal sites using three intra-oral radiographic systems. Additionally, the impact of an automated non-linear grey-level display was evaluated. Ten observers assessed 84 surfaces on bitewing radiographs for their requirement of restorative treatment using a 6-rank confidence scale. Radiographs were taken with conventional film images (Ultraspeed), a storage phosphor plate (Digora) and a CCD system (Dexis). Additionally, the Dexis software was expanded by a contrast enhancement routine (Dexis+). The restorative treatment threshold was defined as presence of macroscopic cavitation. Regarding the areas below mean ROC curves no significant differences were detected between the groups (p>0.05). Likelihood ratios for positive test results were: 5.29 (Ultraspeed), 8.14 (Digora), 9.67 (Dexis) and 11.37 (Dexis+). The accuracy of restorative treatment decisions based on digital and conventional radiographs is comparable. If a dichotomous treatment decision was requested, the digital systems demonstrated a notable tendency towards higher likelihood of true-positive decisions. PMID- 11385200 TI - An intraoral study of caries induced on enamel in contact with fluoride-releasing restorative materials. AB - The aim was to study the effect of three fluoride-releasing restorative materials on sound enamel of approximal teeth under cariogenic conditions intraorally. Bovine enamel slabs were paired with four hemispherical samples from each of three test materials (Ketac Molar, ESPE; Vitremer, F-2000, both from 3M) and one control material (Pertac II, ESPE). Their curved surface touched enamel at a contact point allowing space for plaque accumulation. Pairs were inserted in the buccal flanges of 2 lower dentures for 70 days. Initially and upon dismantling, enamel surface microhardness was measured and the increase in length of indentation in micrometers was recorded at a distance of 0, 0.4, 0.8 and 1.2 mm from the contact point. Respective values were: Pertac II: 16.0+/-7.5, 27.6+/ 15.4, 38.6+/-23.3, 47.8+/-25.2; F-2000: 11.89+/-9.6, 15.8+/-14.2, 21.0+/-15.0, 25.1+/-15.2; Ketac Molar: 6.6+/-6.8, 9.4+/-7.0, 9.5+/-8.0), 13.4+/-9.4; Vitremer: 2.7+/-4.1, 3.5+/-3.5, 3.6+/-4.5), 7.4+/-5.2. Caries protection was Vitremer 82% > Ketac Molar 69% >F-2000 42% in relation to Pertac II (p<0.001). Sections of the slabs examined with polarized light showed typical subsurface lesions only in the Pertac II group, areas of pseudoisotropy in the F-2000 and Ketac Molar groups and no signs of lesion in the Vitremer group. PMID- 11385201 TI - Clinical evaluation of chemomechanical caries removal in primary molars and its acceptance by patients. AB - The purpose of this study was to evaluate the clinical efficiency and patient acceptance of the chemomechanical caries removal agent Carisolvtrade mark in deciduous teeth. Contralateral primary molars of sixteen 7- to 9-year-old patients (32 teeth) were treated with the air-motor and with Carisolv, respectively. Patients replied to a pre- and a postoperative questionnaire. Complete caries removal (CCR) was accomplished in all air-motor sessions. It was not achieved within the time limit of 15 min in 6 (37.5%) Carisolv sessions. Air motor CCR time ranged between 6 and 18 s (mean: 11.81 s ). Carisolv CCR time, where accomplished within 15 min, ranged between 6 min 46 s and 13 min 57 s (mean: 6 min 51 s). Preoperatively, the majority of the children reported disliking the drilling, and that they would visit the dentist more often and prefer sitting in the chair longer if they could avoid it. Postoperatively, the majority of the children reported disliking the taste of Carisolv, estimated Carisolv to have taken longer, would not recommend it to their friends, and preferred the air-motor. In conclusion, Carisolv, although a step forward in terms of solution volume required, is not in a position to replace rotary instruments for caries removal: it did not remove decay completely in one third of our sample; it was much slower than the air-motor; it had a chlorine taste/odor our patients disliked. PMID- 11385202 TI - Effect of time on the remineralisation of enamel by synthetic saliva after citric acid erosion. AB - The aim of this in vitro study was to determine the influence of remineralisation time on rehardening of surface-softened enamel after citric acid erosion. Seven groups of 13 samples of human enamel were eroded in 0.3% citric acid at pH 3.2 for 2 h followed by profilometric measurements. Individual groups of specimens were placed in artificial saliva for 1, 2, 4, 6, 9 or 24 h. A control group was placed in isotonic saline for 24 h. After new profilometric measurements samples were ultrasonicated stepwise up to 480 s with profilometric measurements performed at 5, 30, 120, 240 and 480 s to measure the depth of surface softening. The control group had a softened surface layer of mean thickness 2.9 microm. Mineral deposition was seen at all remineralisation times by scanning electron microscopy. Exposure to artificial saliva for 1, 2 or 4 h produced a partial rehardening of the softened enamel; the additional surface losses produced by ultrasonication were lower and time delayed compared to the control group. Specimens remineralised for 6, 9 and 24 h showed little evidence of surface loss after ultrasonication. The data suggest that a complete rehardening of the softened enamel in vitro is reached after a remineralisation time of 6 h. These data are of clinical relevance to tooth wear. However, there is a need for studies in situ of enamel demineralisation and remineralisation. PMID- 11385203 TI - In situ evaluation of different remineralization periods to decrease brushing abrasion of demineralized enamel. AB - The aim of the present in situ study was to evaluate the effect of different periods of intraoral remineralization to decrease the susceptibility of previously demineralized enamel against toothbrushing abrasion. Six human enamel specimens (A-F) were recessed in the buccal aspects of each of eight intraoral appliances which were worn for 21 days by 8 panelists. Demineralization of the samples was performed twice a day extraorally in the acidic beverage Sprite Light for 90 s. Subsequently, the enamel specimens were brushed at different times. Specimen A was brushed immediately after the demineralization. The remaining samples B-E were brushed after the intraoral appliances had been worn for various periods of remineralization: specimen B, 10 min; C, 20 min; D, 30 min and E, 60 min, respectively. Specimen F was only demineralized and remineralized, but not brushed. After 21 days, enamel wear was measured with a laser profilometer. The following values (mean +/- standard deviation) were obtained: specimen A, 6.78+/ 2.71 microm; B, 5.47+/-3.39 microm; C, 6.06+/-3.18 microm; D, 5.43+/-2.58 microm; E 4.78+/-2.57 microm, and F 0.66+/-1.11 micro;m. Analysis of variance revealed a significant influence of remineralization period on abrasive wear. However, even after a remineralization period of 60 min the wear was significantly increased as compared to the demineralized, but not brushed control. It is concluded that (1) abrasion resistance of softened enamel increases with remineralization period and (2) at least 60 min should elapse before toothbrushing after an erosive attack. PMID- 11385204 TI - Effect of a low-fluoride-content, two-component rinse on fluoride uptake and on de--and remineralization of enamel lesions: an in vitro study. AB - The effect of calcium (Ca) concentrations and added ethanol on fluoride (F) depositions by experimental two-component rinses, each consisting of a Ca containing and an F-containing component, was evaluated in an in vitro system. Among the tested rinses, a 3 mmol/l F two-component rinse with 200 mmol/l Ca and 10% v/v ethanol was found to produce the greatest F deposition relative to the F concentration in the rinse. Specifically, this rinse produced an F deposition that was about 7 times greater than a conventional 13.2-mmol/l sodium fluoride (NaF) rinse. In a second experiment, an in vitro pH cycling model was used to evaluate the potential anti-caries effects of 4 rinses: (1) placebo rinse (no F), (2) 13.2-mmol/l NaF rinse, (3) 52.6-mmol/l NaF rinse, and (4) the 3-mmol/l F two component rinse in a 7-day in vitro pH cycling model. The changes in lesion mineral contents, delta(DeltaZ), as assessed by quantitative microradiographic measurements, were as follows [mean +/- standard deviation, n = 10]: (1) 72.5+/ 10.2 microm, (2) 43.4+/-5.6 microm, (3) 17.3+/-10.2 microm and (4) 45.3+/-5.2 microm. These results showed that the 3-mmol/l F two-component rinse produced the same (p>0.05) protection against demineralization as did the 13.2-mmol/l NaF rinse which had 4 times the fluoride content. The results suggest that it is possible to formulate an effective low-F two-component rinse. PMID- 11385205 TI - Prevention by means of fluoride of enamel erosion as caused by soft drinks and orange juice. AB - Fluoride has been suggested to prevent erosion of the teeth, either after a topical treatment of the teeth or by addition of fluoride to the acidic drink. The main aim of the present study was to describe the dissolution of calcium fluoride in some soft drinks and orange juice and compare it with the amounts of calcium fluoride left on the enamel surfaces after a topical treatment. A further aim was to describe the dissolution of enamel in soft drinks and juice saturated for 3 days with solid calcium fluoride. Solid calcium fluoride was suspended in each of 10 soft drinks and orange juices and gently agitated for 72 h, after which the drinks were analyzed for calcium, phosphate and fluoride and pH was determined. To examine the erosion-preventive effect of the calcium fluoride-rich drink, intact teeth were exposed to the drinks with or without calcium fluoride. It was found that from 6 to 45 mg of calcium fluoride was dissolved per liter of drink. The more acidic the drink, the more calcium fluoride was dissolved, presumably due to HF formation. The teeth exposed to the soft drinks all showed erosion-like lesions. Very little effect of the 4-6 ppm ionic fluoride dissolved in the soft drinks was observed. In orange juice, however, the dissolved calcium fluoride established a saturation with respect to fluorapatite and consequently, the erosion-like lesion was replaced by a caries-like lesion. In conclusion, the acidic soft drinks are capable of dissolving considerable amounts of calcium fluoride and the erosion-preventive effect of even high fluoride concentrations is limited. PMID- 11385206 TI - Platelet glycoprotein IIb/IIIa blockade in acute ischemic stroke. AB - While thrombolytic therapy is approved in the United States and Canada for treatment of acute ischemic stroke, only a small number of patients are currently treated. Additional agents that could restore or improve cerebral flow are needed. Based on the experience in acute coronary syndromes, reperfusion agents such as platelet glycoprotein IIb/IIIa antagonists, alone or in combination with reduced doses of thrombolytic agents, have the potential for improving the safety and efficacy of recombinant tissue plasminogen activator (rt-PA). In addition, these newer agents may permit extension beyond the current 3-hour window from symptom onset used for systemic rt-PA. PMID- 11385207 TI - 4G/5G polymorphism of the plasminogen activator inhibitor-1 gene and insertion/deletion polymorphism of the tissue-type plasminogen activator gene in atherothrombotic stroke. AB - BACKGROUND AND PURPOSE: Decreased fibrinolytic capacity due to increased plasminogen activator inhibitor-1 (PAI-1) activity and decreased tissue-type plasminogen activator (t-PA) activity has been associated with hypertension or atherothrombotic disorders. The aims of this study were to observe associations of the genetic polymorphism for PAI-1 and t-PA with hypertension and atherothrombotic stroke, and to elucidate whether impaired fibrinolytic activity in atherothrombotic stroke was related to atherothrombosis per se or to other risk factors such as hypertension. METHODS: Patients with atherothrombotic stroke (n = 60), hypertension (n = 100), and control subjects (n = 100) were enrolled. We genotyped all subjects for 4G/5G polymorphism in the promoter region of the PAI-1 gene and the Alu-repeat insertion/deletion (I/D) polymorphism in intron h of the t-PA gene by polymerase chain reaction and endonuclease digestion. RESULTS: The frequency of the 4G/4G genotype of PAI-1 was significantly higher in the atherothrombotic stroke patients than the control subjects (41.7 versus 21%; p = 0.005), but not in the hypertensive subjects. There was a significant association between 4G/4G genotype of PAI-1 and atherothrombotic stroke (adjusted odds ratio = 3.11, 95% confidence interval 1.18-8.15), adjusting for age, sex, total cholesterol, low-density lipoprotein, triglyceride, and body mass index. However, the number of the I/I genotype of t-PA in the atherothrombotic stroke or hypertensive patients was virtually identical to the control subjects. CONCLUSION: Our results suggest that the 4G/4G genotype of the PAI-1 gene is significantly associated with an increased risk of atherothrombotic stroke. This finding also supports that impaired fibrinolytic activity in atherothrombotic stroke is related to atherothrombosis per se, but not to hypertension, one of the most important risk factors of atherothrombotic stroke. PMID- 11385208 TI - Intracarotid abciximab injection to abort impending ischemic stroke during carotid angioplasty. AB - BACKGROUND: Abciximab, a platelet glycoprotein IIb/IIIa receptor antagonist, prevents ischemic complications during percutaneous transluminal coronary angioplasty and was recently shown to open occluded vessels in patients with acute myocardial infarction when used alone or in combination with other thrombolytic agents. In an animal model of arterial thrombosis, abciximab was found to be safe and effective for the prevention of carotid artery thrombosis. However, the safety and efficacy of abciximab in the treatment of acute ischemic cerebrovascular events is unknown at present. CASE DESCRIPTION: We describe 3 patients who experienced ischemic cerebrovascular events with symptoms involving the middle cerebral artery territory while undergoing percutaneous angioplasty and stenting to their internal carotid arteries. Abciximab was administered to each patient within 10 min of symptom onset as a bolus (0.25 mg/kg) into the ipsilateral common carotid artery followed by continuous intravenous infusion (9 microg/min) for 12 h. All patients' symptoms resolved completely (by 25 min, 40 min and 5 h, respectively) with no further neurological complications. CONCLUSIONS: Our preliminary observation suggests that abciximab may improve neurological outcome following middle cerebral artery ischemic events associated with carotid angioplasty and stenting. Large prospective studies are warranted to establish the safety and efficacy of abciximab in acute ischemic stroke, either as a primary treatment modality or an adjunct to carotid angioplasty and stenting. PMID- 11385209 TI - Validity and reliability of simple questions in assessing short- and long-term outcome in Norwegian stroke patients. AB - The utility of simple questions for the assessment of stroke outcome in large scale international studies has generally been approved, but their validity and reliability have not been evaluated in different cultures or at different intervals after a stroke. The study comprised 150 stroke patients who had been admitted consecutively to a stroke unit 6 weeks or 6 months earlier. Two weeks before the visit the patient received a postal questionnaire containing the simple 'dependency' question: 'In the last 2 weeks, did you require help from another person for everyday activities?' and the simple 'recovery' question: 'Do you feel that you have made a complete recovery from your stroke?'. The visit was performed by trained personnel unaware of the patient's or his carer's replies, and comprised the same 2 questions administered by the personnel, the Barthel ADL Index (BI) and the modified Rankin Scale (mRS). The patients' functional status was categorised as good or bad according to the chosen cutoff levels on BI and mRS. At 6 months the dependency question had an accuracy of 83 and 82% in identifying patients with good or bad outcome, defined as BI > or = 95 or < 95 and mRS < 3 or > or = 3, respectively, whereas the recovery question had an accuracy of 86% when compared with mRS = 0 or > 0. There was no difference in accuracy of the simple questions at 6 weeks compared with 6 months, and there was no clinically important difference between responses from patients and carers. The agreement between the responses to the questionnaire and the interview was good to moderate (kappa = 0.62 for the dependency question, and 0.55 for the recovery question). We conclude that the simple questions seem to be valid and reliable measures of stroke outcome when tested in Norwegian patients after 6 weeks or 6 months, which supports their continued use in large-scale multinational stroke studies at different intervals after stroke. PMID- 11385210 TI - Differences between anterior and posterior circulation stroke in TOAST. AB - BACKGROUND AND PURPOSE: Clinicians have tended to view anterior circulation (AC) and posterior circulation (PC) strokes as separate entities, with different underlying pathogenesis, natural histories, and potential responsiveness to interventions such as anticoagulation. We sought to explore differences between AC and PC stroke in the Trial of ORG 10172 in Acute Stroke Treatment (TOAST). METHODS: For patients enrolled in TOAST, prospective clinical information was collected including outcome at 3 months. Data on vascular distribution were obtained from the clinical impression of the investigators. Group comparisons for categorical data were performed using Fisher's exact test. Independent sample t tests and analysis of covariance were used for all continuous data. RESULTS: The analysis included 1,039 patients with AC stroke and 180 patients with PC stroke. There were fewer women in the PC than in the AC groups, but otherwise there were no differences in demographics, risk factors or stroke subtypes between the two groups. Headache (AC 8.7%, PC 15%, p = 0.013) and vomiting (AC 3.5%, PC 17.8%, p < 0.001) were more common among PC patients. Mean baseline National Institutes of Health Stroke Scale (NIHSS) score was lower (less severe) among PC (6.1) than AC patients (9.5; p < 0.001). On univariate analysis, favorable outcome at 3 months was more common for PC patients in both the placebo group (PC 82%, AC 71%, p = 0.04) and heparinoid group (PC 87%, AC 73%, p = 0.005). However, multivariate analysis, controlling for gender, history of previous stroke and baseline NIHSS score, showed no difference in outcome between PC and AC stroke. For favorable outcome, there was no interaction between vascular distribution and treatment category, suggesting that the effect of heparinoid did not differ between PC and AC strokes. CONCLUSION: Patients with PC stroke seem to have a better long-term outcome than do AC patients, but this difference is no longer apparent when controlling for important prognostic variables. PC patients did not show any particular benefit from anticoagulation, and the efficacy of heparinoid did not vary between AC and PC stroke. While AC and PC patients do differ in some respects, it may be inappropriate to single out PC patients for anticoagulant treatment. PMID- 11385212 TI - Acetylsalicylic acid and microembolic events detected by transcranial Doppler in symptomatic arterial stenoses. AB - BACKGROUND: In patients with symptomatic carotid artery stenosis, high-intensity transient signals detected by transcranial Doppler (TCD) have been related to particulate microemboli originating at the stenotic lesion. The occurrence of these microembolic events within the Doppler spectrum should be influenced by antithrombotic agents of proven efficacy in these patients mainly by reducing cerebral embolism. METHODS: Seventy-four of 192 consecutive patients with symptomatic arterial stenosis in the anterior circulation and clinical symptoms within the last 30 days underwent 1-hour bilateral TCD monitoring. Patients were selected, if they presented temporal bone windows enabling transcranial insonation, revealed normal Doppler CO2 test excluding hemodynamic impairment, had not received antithrombotic therapy other than acetylsalicylic acid (ASA) before sonographic examination, and gave informed consent to 1-hour monitoring which could be performed immediately on admission/presentation of the patient at the Department of Neurology. RESULTS: Microembolic events were detected in 38 patients (51%). The proportion of patients with events among 26 patients without antithrombotic medication was 73% as compared with 40% in 48 patients receiving ASA at the time of TCD monitoring (p = 0.023). Multivariate analysis including time from ischemia to TCD, presence and start of ASA prevention, degree and localization of stenosis, and presence of a single or recurrent ischemia revealed that absence of an ASA prevention (odds ratio OR 7.1, 95% confidence interval CI 1.6-31.4, p = 0.010), recurrent ischemic events (OR 7.1, 95% CI 1.6-32.7, p = 0.011), and extracranial localization of the stenosis (OR 3.8, 95% CI 1.1-13.2, p = 0.038) were independent predictors for microembolic events. CONCLUSION: In patients with symptomatic arterial stenosis, the absence of an ASA medication is associated with the occurrence of TCD-detected microembolic events, suggesting a relation between these events and ASA-sensitive microemboli from the stenotic lesion. PMID- 11385211 TI - Diagnostic impact of early transcranial Doppler ultrasonography on the TOAST classification subtype in acute cerebral ischemia. AB - OBJECTIVE: The impact of early transcranial Doppler ultrasonography (TCD) upon stroke subtype diagnosis is unknown and may affect therapeutic strategies. In this study, the diagnostic usefulness of TCD in stroke subtype diagnosis according to the criteria of the Trial of ORG 10172 in Acute Stroke Treatment (TOAST) study was investigated in patients with acute cerebral ischemia. METHODS: TCD examination within 24 h of symptom onset was performed in 50 consecutive patients with acute cerebral ischemia. Of these 54% were female. Sixty percent of patients were black, 36% white, and 4% Asian. Initial TOAST stroke subtype diagnosis (ITSSD) was based upon clinical presentation and initial brain imaging studies. Modified TOAST stroke subtype diagnosis was determined subsequently after additional review of the TCD examination. Final TOAST stroke subtype diagnosis was determined at hospital discharge, incorporating all diagnostic studies. Using final TOAST stroke subtype diagnosis as the 'gold standard' ITSSD and modified TOAST stroke subtype diagnosis were compared in order to determine additional benefit from the information obtained by TCD. Data were collected retrospectively by a single investigator. RESULTS: ITSSD classified 23 of 50 (46%) patients correctly. After TCD, 30 of 50 (60%) patients were classified correctly, for an absolute benefit of 14% and a relative benefit of 30% (p = 0.018). Most benefit from TCD was observed in the TOAST stroke subtype category large-artery atherosclerosis, in particular in patients with intracranial vascular disease. In this category, ITSSD had a sensitivity of 27% which increased to 64% after TCD (p = 0.002). CONCLUSION: TCD within 24 h of symptom onset improves the accuracy of early stroke subtype diagnosis in patients with acute cerebral ischemia due to large-artery atherosclerosis. This may have clinical implications for early therapeutic interventions. PMID- 11385213 TI - HITS in internal carotid dissections. AB - Carotid dissections can cause neurological deficits either by hemodynamic or embolic mechanisms. Anticoagulants are often used in this setting to prevent neurological deterioration and stroke recurrence. High-intensity transient signals (HITS) detected by transcranial Doppler may be due to microembolism. We investigated the presence and number of HITS in a series of 27 patients with carotid dissection. HITS were detected in 14 (52%) patients and only in cases with a less than 7-day evolution. No association was found between HITS and clinical or imagiological features of the dissection nor with neurological aggravation or treatment type. We failed to demonstrate any relevant clinical significance of HITS in carotid dissection. PMID- 11385214 TI - Accuracy of color-Doppler in the quantification of proximal vertebral artery stenoses. AB - BACKGROUND: Vertebrobasilar (VB) strokes appear to have the same causes as carotid strokes. Obstructive lesions of proximal vertebral arteries probably occur in about 30% of stroke patients. PURPOSE: Our aim was to assess the validity of color Doppler sonography compared to selective intra-arterial angiography in the quantification of proximal vertebral artery stenoses. MATERIALS AND METHODS: A prospective blind study of 316 vertebral arteries was undertaken between 1996 and 1998. One hundred and fifty-eight patients with cerebrovascular disorders without cerebral hemorrhage were studied consecutively by frequency or amplitude color Doppler flow imaging and intra-arterial angiography. The lesions were quantified by morphological and hemodynamic criteria and classified into 6 groups: 0% 207 arteries; 1-29% 32 arteries; 30-49% 29 arteries; 50-69% 13 arteries; 70-99% 23 arteries; 100% 12 arteries. RESULTS: Ten of the 12 occlusions were identified, the 2 false-negatives were due to 2 revascularized vessels. Moderate stenoses (<50%) were differentiated from tight stenoses (>50%) using hemodynamic criteria. The majority of false-negative stenoses (38) in the different groups were related to intrathoracic or very deep origin of the artery, anechogenic stenosis or a tortuous vessel. Stenoses greater than 70% were diagnosed in 71% of cases with a specificity of 99%. The kappa value was 0.80. CONCLUSION: Duplex sonography should be proposed first in VB attacks or stroke to detect and quantify vertebral artery stenoses for surgery and angioplasty. PMID- 11385215 TI - MR angiographic evaluation of carotid and intracranial arteries in Japanese patients scheduled for coronary artery bypass grafting. AB - BACKGROUND AND PURPOSE: Stroke is a serious complication of coronary artery bypass grafting (CABG). Preoperative evaluation of the cerebral arteries to identify patients at increased risk of stroke after CABG is important. In a prospective study, we evaluated cerebral artery occlusive lesions with MR angiography in Japanese patients scheduled to undergo CABG to determine the prevalence of occlusive diseases in the extracranial carotid and intracranial arteries in this population and to identify preoperative risk factors for these patients. METHODS: The subjects were 151 consecutive patients (115 men and 36 women ranging in age from 41 to 82 years) who were scheduled for CABG under nonemergency conditions between October 1995 and February 1998. Carotid and intracranial arteries were examined for occlusive lesions with MR angiography. Patient demographics and risk factors including age, sex, hypertension, diabetes mellitus, hyperlipidemia, smoking habit, history of stroke, peripheral vascular disease and preoperative thromboembolic infarcts revealed by MR imaging were recorded and analyzed. RESULTS: Cervical carotid artery stenoses of more than 50% narrowing were detected in 16.6% of the subjects, and intracranial artery stenoses of more than 50% narrowing were detected in 21.2% of the subjects. Multiple logistic regression analyses identified peripheral vascular disease and lacunar infarcts in the basal ganglia as significant and independent predictors of cervical carotid arterial stenoses. No significant predictor for intracranial arterial stenoses was identified. CONCLUSIONS: The prevalence of extracranial carotid and intracranial artery stenosis in Japanese patients scheduled for CABG is considerably high. MR angiography is of value of identifying these patients. Preoperative evaluation of cranial arteries is recommended, particularly in patients with peripheral vascular disease and infarcts in the basal ganglia. PMID- 11385216 TI - Marfan's syndrome and multiple extracranial aneurysms. AB - Dissecting aneurysms of the extracranial cervical arteries in Marfan's syndrome are most frequently caused by the extension of aortic dissection. Spontaneous and isolated aneurysms limited to the major extracranial cervical arteries have also been reported in Marfan's syndrome, although very rarely. In these cases, the aneurysms are usually asymptomatic or may present as a pulsatile mass. Transient ischemic attacks (TIAs) or strokes, associated with isolated extracranial aneurysms, have not been mentioned in Marfan's syndrome. We report a patient with Marfan's syndrome presenting with TIAs in whom we detected a saccular aneurysm of the vertebral artery as well as bilateral fusiform aneurysms of the internal carotid arteries. PMID- 11385217 TI - Detection of acute pontine infarction by diffusion-weighted MRI in capsular warning syndrome. PMID- 11385218 TI - Meningovascular syphilis: a vascular syndrome with typical features? PMID- 11385219 TI - Stroke prevention in atrial fibrillation among Hong Kong Chinese. PMID- 11385220 TI - Bilateral thalamic stroke due to neurocysticercosis in a non-endemic area. PMID- 11385221 TI - High-dose pulse corticosteroid therapy: is it indicated for severe alopecia areata? PMID- 11385222 TI - Insect bite reactions: an update. AB - Contact with insects may induce cutaneous or systemic reactions, ranging from little more than annoying to life threatening. The diagnosis depends on maintaining a high index of suspicion and being familiar with the insect fauna of the patient's environment and area of travel. PMID- 11385223 TI - Identification of TIA-1+ and granzyme B+ cytotoxic T cells in lichen sclerosus et atrophicus. AB - BACKGROUND: The onset and persistence of cutaneous lichen sclerosus et atrophicus (LSA) are linked to the presence of an inflammatory infiltrate of CD3+ T cells that includes CD4+ and CD8+ cells. The functional relevance of the presence of these cells is unknown. OBJECTIVE: The study intended to quantify resting and activated cytotoxic T cells in LSA lesions. METHODS: Twenty patients with active LSA were studied. Skin-infiltrating T cells were immunohistologically characterized with antibodies against CD3, CD8, T-cell-restricted intracellular antigen (TIA-1) and granzyme B (GrB). TIA-1 labels cytotoxic granules of resting and activated T cells, whereas GrB designates activated cytotoxic T lymphocytes (CTL). RESULTS: In all cases, numerous T cells were consistently found expressing cytotoxic granules. The results indicated a high number of infiltrating CD8+ TIA+ T cells. Furthermore, a notable number of GrB+ activated CTL associated with hydropic degeneration of the basal cell layer were found within the dermal infiltrate and at the dermoepidermal interface. CONCLUSION: This study shows that a high proportion of skin-infiltrating T cells in LSA has a potential cytotoxic function. The results indicate that hydropic degeneration of basal keratinocytes may at least partially be mediated by CTL-dependent mechanisms. Our data also indicate that a cell-mediated immune response may play an important role in the pathogenesis of the disease. PMID- 11385224 TI - Epidermal growth factor receptor overexpression and genetic aberrations in metastatic squamous-cell carcinoma of the skin. AB - BACKGROUND: Cutaneous squamous-cell carcinoma (SCC) sometimes causes lymph node metastasis and results in poor prognosis. However, little is known about cytogenetic alterations underlying tumor progression or metastasis. The aim of the present study was to investigate the genetic aberrations and expression of epidermal growth factor receptor (EGFR) in metastatic SCC of the skin. METHODS: We undertook comparative genomic hybridization (CGH) analysis of 4 specimens which were obtained from a case of cutaneous SCC, including the primary lesion and 3 lymph nodes of the metastatic lesion. RESULTS: Only one amplified locus (7p12-13) was detected in any metastatic lymph node, in which the EGFR gene is located. Therefore, we applied immunohistochemistry for EGFR to 5 cases of metastatic SCC including the case analyzed using CGH and 4 other cases (5 primary and 5 metastatic lesions). EGFR was expressed in 4 of 5 cases (both primary and metastatic lesions, including the case analyzed using CGH), and the staining patterns of primary and metastatic lesions were different. The primary tumors were focally weakly positive for immunostaining (+), whereas the 4 metastases were diffusely and strongly positive (+++). CONCLUSIONS: Our findings suggest that the clone with EGFR expression might selectively metastasize in some cutaneous SCCs. The existence of an EGFR-negative case reveals that EGFR expression is not always required for skin carcinogenesis, but expression of EGFR might confer metastatic potential of cutaneous SCCs. PMID- 11385225 TI - Microdialysis can detect age-related differences in glucose distribution within the dermis and subcutaneous adipose tissue. AB - BACKGROUND: Intrinsic (chronological) cutaneous aging is a continuous and complex process. Limitation in nutrient supply to the skin could contribute to this process. OBJECTIVE: Our study focused on the impact of age on glucose supply to human skin and its distribution within the dermis and subcutaneous adipose tissue. METHODS: An oral glucose load (100 g) was given to healthy volunteers of two age groups (24 +/- 4 and 59 +/- 10 years, n = 4 for each group). Dialysate concentrations of glucose and glycerol were monitored by microdialysis in the dermis and adipose tissue. RESULTS: Baseline tissue [glucose] did not differ significantly between the two groups. After the glucose load, dialysate [glucose] of the dermis peaked between 60 and 80 min and between 80 and 120 min in the young and old groups, respectively. In the old group, dialysate [glucose] was significantly higher in the dermis than adipose tissue at 80, 100, 120, 160 and 180 min after the load. The sum of the areas under the curve (dermis + adipose tissue) did not differ significantly between the two age groups. CONCLUSIONS: Aging is accompanied by an increased diffusion barrier for glucose between the dermis and adipose tissue. PMID- 11385226 TI - The heterogeneous clinical spectrum of genital herpes. AB - BACKGROUND AND OBJECTIVE: Most data of genital herpes have been collected in STD clinics in the USA where unrecognized forms accounted for 80% of HSV-2 infections. Our aim was to study the clinical features in an outpatient clinic of dermatology. METHODS: The charts of 170 patients, previously monitored prospectively for a HIV prevalence study, with culture-confirmed genital herpes or herpetic infection with HSV-2 at any other site presenting between 1995 and 1999 were analyzed. RESULTS: 111 (65%) men and 59 (35%) women were identified with a mean age of 44 years. Only 49% had a typical cluster of genital lesions. Eighty-six (51%) presented with either lesions at extragenital sites [mostly the buttocks 33/170 (19%), thigh 10/170 (6%), anal region 9/170 (5%) and fingers 8/170 (5%)] or showed morphologically atypical forms of isolated genital lesions [single ulcer 16/170 (9%), erosion 6/170 (4%), crust 3/170 (2%) and fissure, edema or erythema each 1/170 (1%)]. Women significantly presented more often with extragenital infections of HSV-2 [36/59 (61%)] than men [18/111 (16%)]. CONCLUSIONS: More than half of the patients with genital herpes of a mainly immunocompetent population presented with atypical manifestations. The underdiagnosis of genital herpes seems largely due to misinterpretation of atypical genital and extragenital lesions. PMID- 11385227 TI - Onychomycosis and other superficial fungal infections of the foot in the elderly: a pan-European survey. AB - BACKGROUND: The escalating ageing population in the western world has led to an increased incidence of superficial fungal infections. The most common infections include onychomycosis, tinea pedis and tinea cruris. With the increasing life expectancy, the prevalence of onychomycosis and other superficial fungal infections is likely to increase further without adequate prevention and treatment. OBJECTIVE: To study the prevalence of foot mycoses in Europe. METHOD: The Achilles project represents a survey of 90,085 subjects from 16 European countries. RESULTS: Approximately half of the total screened population had evidence of fungal foot infection, with tinea pedis and onychomycosis affecting one quarter of these individuals. Advancing age showed an unfavourable effect on the prevalence of these infections. CONCLUSION: As the number of aged people in developed countries continues to increase, skin diseases will constitute a greater pharmaco-economic concern of worldwide healthcare. Better recognition by clinicians and patients of mycotic foot disease will help prevent direct morbidity and further complications. PMID- 11385228 TI - Prognostic and therapeutic implications of sentinel lymphonodectomy and S-staging in Merkel cell carcinoma. AB - BACKGROUND: Merkel cell carcinoma (MCC) is a rare but very aggressive neuroendocrine neoplasm of the skin with a high propensity for early lymph node metastasis and subsequent distant spread. Optimal treatment and prognostic factors are poorly defined. OBJECTIVE: The purpose of this study is to assess the prognostic and therapeutic relevance of sentinel lymphonodectomy in MCC. METHODS: Five patients with biopsy-proven MCC underwent gamma-probe-guided sentinel lymphonodectomy assisted by lymphatic mapping. From each sentinel lymph node (SLN), a series of paraffin sections was histologically and immunohistochemically examined for the presence of micrometastases, which were then staged according to the recently published S classification. RESULTS: Four of the 5 patients showed metastatic disease in the SLNs, 3 of which were classified as S(2), 1 as S(3). Only 1 of the S(2) patients demonstrated additional positive nodes at completion lymphonodectomy. The patient staged as S(3) refused a radical lymph node dissection and died within 1 year due to widespread metastasis. CONCLUSIONS: Sentinel lymphonodectomy is a low-morbidity procedure which enables an early detection and exact staging of regional lymph node metastases with potentially high prognostic and therapeutic relevance in MCC. PMID- 11385229 TI - High-dose pulse corticosteroid therapy in the treatment of severe alopecia areata. AB - OBJECTIVE: The present monocenter prospective study was designed to evaluate the efficacy of intravenous high-dose methylprednisolone pulse therapy in patients with severe alopecia areata (AA). METHODS: 30 patients (aged 14-56 years) were treated with methylprednisolone (8 mg/kg body weight) intravenously on 3 consecutive days at 4-week intervals for at least 3 courses. RESULTS: 67% of patients with AA plurifocalis showed >50% regrowth of hair. None of the patients with AA totalis or universalis and only 1 patient with ophiasic AA responded to therapy. In patients with AA plurifocalis, higher response rates could be observed in those suffering from long-term disease compared to patients treated during their first episodes of AA (73 vs. 57%). CONCLUSION: High-dose methylprednisolone pulse therapy is an effective and well-tolerated treatment for patients with severe AA plurifocalis but might be less beneficial for patients with ophasic AA, AA totalis or universalis. PMID- 11385230 TI - Efficacy of itraconazole, terbinafine, fluconazole, griseofulvin and ketoconazole in the treatment of Scopulariopsis brevicaulis causing onychomycosis of the toes. AB - BACKGROUND: Scopulariopsis brevicaulis is a common non-dermatophyte mould that can cause onychomycosis. OBJECTIVE: To evaluate the efficacy and safety of the oral antifungal agents griseofulvin, ketoconazole, itraconazole, fluconazole and terbinafine in the treatment of S. brevicaulis. PATIENTS AND METHODS: In a prospective, comparative, parallel-group, single-blinded, randomized, non industry-sponsored study, patients with toe onychomycosis caused by S. brevicaulis sp. were randomized and treated with one of 5 oral antifungal agents, i.e. griseofulvin, ketoconazole, itraconazole (pulse), fluconazole or terbinafine. The treatment regimens were: griseofulvin 600 mg twice daily for 12 months, ketoconazole 200 mg daily for 4 months, itraconazole pulse therapy given for 3 pulses, with each pulse consisting of 200 mg twice daily for 1 week with 3 weeks off between successive pulses, terbinafine 250 mg daily for 12 weeks and fluconazole 150 mg daily for 12 weeks. RESULTS: There were 59 patients (48 males, 11 females, mean age 35.6 years, range 25-53 years). All patients had clinical evidence of distal and lateral onychomycosis, with moderate to severe disease of the target nail. Between the treatment groups there was no significant difference in the mean age of the patients or the mean area of involvement with onychomycosis at baseline. The efficacy parameters were clinical cure (CC) and mycological cure (MC). At month 12 after the start of treatment, the response was: griseofulvin, CC 3/11, MC 0/11, CC + MC 0/11; ketoconazole, CC 10/12, MC 8/12, CC + MC 8/12; itraconazole, CC 12/12, MC 12/12, CC + MC 12/12; terbinafine, CC 12/12, MC 11/12, CC + MC 11/12, and fluconazole, CC 8/12, MC 8/12, CC + MC 8/12. Adverse effects consisted of: griseofulvin, gastro-intestinal symptoms, allergic reaction, photodermatitis, hepatic and renal dysfunction in 11 patients with discontinuation of treatment in 3 patients; ketoconazole, hepatic dysfunction but no symptomatic changes in 2 patients; itraconazole, nausea and vomiting in 2 patients; terbinafine, taste disturbance in 2 patients, nausea in 3 patients, and fluconazole, severe gastro-intestinal events in 5 patients. None of the patients receiving ketoconazole, itraconazole, terbinafine or fluconazole discontinued treatment. CONCLUSIONS: Itraconazole and terbinafine demonstrate efficacy against some cases of S. brevicaulis toe onychomycosis. These agents also appear to be safe in the course of therapy for toe onychomycosis. Griseofulvin is ineffective against toe onychomycosis caused by S. brevicaulis. Ketoconazole is not recommended for toe onychomycosis given its potential for adverse effects, particularly with the availability of the newer antifungal agents. PMID- 11385231 TI - Interferon-gamma-induced RANTES production by human keratinocytes is enhanced by IL-1beta, TNF-alpha, IL-4 and IL-13 and is inhibited by dexamethasone and tacrolimus. AB - BACKGROUND AND METHODS: Recent studies have shown that RANTES plays a role in the pathogenesis of inflammatory skin diseases. We examined the production of RANTES by human keratinocytes (KCs) when cultured with various cytokines. RESULTS: IFN gamma (100 ng/ml) or IL-1beta (100 ng/ml) significantly induced RANTES production by KCs in 48-hour culture. These cytokines synergistically increased RANTES production by KCs. TNF-alpha (100 ng/ml), IL-4 (100 ng/ml) or IL-13 (100 ng/ml) markedly enhanced the RANTES production by KCs induced by IFN-gamma (100 ng/ml) although none of those cytokines significantly enhanced that induced by IL-1beta (100 ng/ml) in 48-hour culture. Dexamethasone (10(-8) M) strongly inhibited RANTES production by KCs induced by the combination of IFN-gamma and IL-4, while tacrolimus (FK-506, 10(-8) and 10(-6) M) showed partial inhibition. CONCLUSIONS: These results revealed that RANTES production by KCs is regulated by inflammatory cytokines, such as IFN-gamma, IL-1beta, TNF-alpha, IL-4 and IL-13, and can be modulated by immunosuppressive drugs. Our data suggest that RANTES is involved in skin inflammation. PMID- 11385232 TI - Didymosis aplasticosebacea: coexistence of aplasia cutis congenita and nevus sebaceus may be explained as a twin spot phenomenon. AB - BACKGROUND: Co-occurrence of aplasia cutis congenita and nevus sebaceus has sometimes been observed. OBJECTIVE: We propose a genetic mechanism that may explain this coexistence of two different congenital skin disorders. METHODS AND RESULTS: We review 15 cases as reported in the literature and present an additional case. To explain the temporal and spatial proximity of the two skin lesions we advance the concept of twin spotting for which we propose the term didymosis (Greek didymos = twin). In an embryo heterozygous for two different recessive mutations localized on either of a pair of homologous chromosomes, an event of somatic recombination would occur at an early developmental stage, giving rise to two different cell clones homozygous for either mutation. CONCLUSION: The concept of didymosis aplasticosebacea is so far hypothetical. Future molecular research may show whether this concept holds true. PMID- 11385233 TI - A case of giant extramammary Paget's disease of the genital area with squamous cell carcinoma. AB - Extramammary Paget's disease is a primary carcinoma with apocrine differentiation that begins within the epidermis. Extramammary Paget's disease and squamous-cell carcinoma (SCC) have different neoplastic origins and are different both clinically and histologically. Until now the concurrence of extramammary Paget's disease and SCC in the same area has not been reported to our knowledge. We report on a 76-year-old man with extramammary Paget's disease affecting a large region of the genital area with SCC within the same area. A skin biopsy specimen showed the coexistence of typical Paget's disease and highly differentiated SCC invading the dermis. Immunohistochemical staining of both cell types was completely different. PMID- 11385234 TI - Grover's disease associated with Sarcoptes scabiei. AB - An 83-year-old man presented with a 4-month history of discrete, itchy papules mainly distributed on the trunk and upper extremities. Histopathologic examination of two biopsies from lesions on the trunk revealed mainly focal suprabasal acantholysis and an inflammatory infiltrate composed mainly of lymphocytes with a few eosinophils. The overall clinical and histopathologic features were consistent with Grover's disease. However, scrapings taken from the skin lesions showed numerous mites of Sarcoptes scabiei. Subsequent treatment with an antiscabies cream led to a rapid complete cure, and no skin lesions have been observed during a 6-month follow-up. A review of the literature revealed 2 other cases of cutaneous lesions fulfilling the clinical and histopathologic features of Grover's disease in which mites of S. scabiei were demonstrated. Our observation further highlights the unusual association of Grover's disease with S. scabiei mites and emphasises the importance of excluding this easily treatable skin infestation in all patients with Grover's disease. PMID- 11385235 TI - Hyperkeratosis lenticularis perstans (Flegel's disease)--lack of response to treatment with tacalcitol and calcipotriol. AB - Hyperkeratosis lenticularis perstans (HLP) or Flegel's disease is a rare dermatosis characterized by asymptomatic hyperkeratotic papules predominantly located on the lower extremities. Lesional and non-lesional epidermis samples were studied by light- and electron-microscopic examination. The main ultrastructural finding was the presence of structurally altered Odland bodies/membrane-coating granules. Different therapeutic options for HLP have been reported, but none of the treatments was shown to be consistently effective. Here, we report on a patient with Flegel's disease who did respond to topical 5 fluorouracil, whereas topical vitamin D(3) synthetics were ineffective. PMID- 11385236 TI - Syndactyly/brachyphalangy and nail dysplasias as marker lesions for sclerosteosis. AB - Sclerosteosis describes an autosomal recessive form of hyperostosis corticalis generalisata (MIM 239100). Sclerosteosis is primarily a disorder of osteoblast hyperactivity and metabolic abnormalities are not present. Besides generalized bone changes the presence of asymmetric cutaneous syndactyly of the index and middle fingers is characteristic. In many cases this syndactyly is associated with nail dysplasia and therefore dermatologists should recognize this clinical finding as a possible marker of this entity. We report on a 36-year-old female of Greek origin who had had finger and nail dysplasias and facial asymmetry since birth. The patient was hospitalized on the Neurology ward because of increasing spastic and ataxic gait disturbances. Physical examination revealed numerous neurological problems resulting from bony compression of nerves. Furthermore the patient had remarkably deformed fingers with hypoplasia of the second finger on both sides. The nails were dysplastic, especially on both index fingers. All laboratory results concerning metabolic diseases were normal. It has been shown that sclerosteosis is clinically and radiographically very similar to van Buchem disease. Through a genome-wide search with a highly polymorphic microsatellite the gene responsible for van Buchem disease has been mapped to 17q12-q21, and Balemans et al. (1999) assigned the locus for sclerosteosis to the same region providing genetic support for the hypothesis of allelism. Dermatologists should be able to interpret such syndactyly associated with nail deformities as a possible hint for the diagnosis of sclerosteosis in patients with hyperostotic features. PMID- 11385237 TI - Basal cell carcinoma in association with multiple trichoepitheliomas. AB - We report the case of a basal cell carcinoma (BCC) associated with multiple trichoepitheliomas (TEs) and include the results of histopathological and immunohistochemical studies. The patient had a large, pigmented nodule associated with 4 flesh-colored papules in the central facial area. Two of the 4 flesh colored papules and the large pigmented nodule were excised. Histopathologically, the pigmented nodule was diagnosed as BCC and 2 of the papular lesions were diagnosed as TEs. In both the BCC and TEs, almost all of the basaloid aggregations were diffusely positive for cytokeratin (CK) 19, CK8 and CK8/18. Based on these clinicopathological observations and reports in the recent literature, these two neoplasms are considered to be highly related. BCC appears to differentiate toward follicular germinative cells, and TE is its benign counterpart. PMID- 11385238 TI - Mucosal leishmaniasis occurring in a renal transplant recipient. PMID- 11385239 TI - Alternariosis refractory to itraconazole in a patient suffering from bullous pemphigoid. PMID- 11385240 TI - Unilateral nodular elastosis with cysts and comedones (Favre-Racouchot syndrome). PMID- 11385241 TI - Multiple anemic macules on the arms: not a variant form of nevus anemicus. PMID- 11385242 TI - Multiple anemic macules or Bier's spots? PMID- 11385244 TI - Sezary syndrome with acute progressive pulmonary infiltration after improvement of skin eruption. PMID- 11385247 TI - Update on the pathology and genetics of exocrine pancreatic tumors with ductal phenotype: precursor lesions and new tumor entities. AB - The majority of pancreatic neoplasms show a ductal phenotype and can be classified as ductal adenocarcinomas. Pancreatic duct lesions have been discussed as tumor precursors. This review presents a recently adopted standard system for these lesions which distinguishes among three grades of pancreatic intraepithelial neoplasia (PanIN). Molecular studies revealed that PanIN-2 and PanIN-3 lesions represent a distinct step towards invasive carcinoma. Another focus of the review is the advances that have been made in the further immunohistochemical and molecular characterization of special pancreatic neoplasms showing a ductal phenotype, such as undifferentiated carcinoma, mucinous noncystic (colloid) carcinoma, intraductal papillary mucinous neoplasm, mucinous cystic neoplasm, medullary carcinoma, and other rare tumors. PMID- 11385246 TI - Epidemiological trends in pancreatic neoplasias. AB - Primary prevention is the most effective approach to reduce the incidence of pancreatic cancer. Epidemiological studies have contributed to the identification of risk factors for pancreatic cancer, suggesting an association with age, various medical conditions, environmental and lifestyle risk factors, and occupational and genetic conditions. Age is the strongest risk factor. The most consistently identified environmental risk factor is smoking, but there is less certainty concerning dietary factors. Studies have suggested a positive association with high energy intake, cholesterol and meat, while vegetable and fruit intakes are probably protective. Patients with chronic pancreatitis and new onset of diabetes mellitus have a low but increasing risk of having or developing pancreatic cancer. There is strong evidence for the association of hereditary pancreatitis or cystic diseases of the pancreas and pancreatic cancer. A family history of pancreatic cancer is an important risk factor, but only a small proportion can be linked with known familial cancer syndromes. Thus, additional yet unidentified predisposing risk factors have to be assumed. PMID- 11385248 TI - Experimental animal models in pancreatic carcinogenesis: lessons for human pancreatic cancer. AB - The silent course of pancreatic cancer and its explosive fatal outcome have hindered studies of tumor histogenesis and the identification of early biochemical and genetic alterations that could help to diagnose the disease at a curable stage and develop therapeutic strategies. Experimental animal models provide important tools to assess risk factors, as well as preventive and therapeutic possibilities. Although several pancreatic cancer models presently exist, only models that closely resemble human tumors in morphological, clinical, and biological aspects present useful media for preclinical studies. Because an estimated 70% of human tumors are induced by carcinogens and because a significant association has been found between cigarette smoking and pancreatic cancer, chemically induced models are of particular value. Moreover, in such models the etiology, modifying factors, effects of diets, and naturally occurring products can be studied and early diagnostic, preventive, and therapeutic possibilities sought out. Many of the existing models are described in this review, and the advantages and shortcomings of each model and their clinical implications are discussed. PMID- 11385249 TI - Differentiation of chronic pancreatitis from pancreatic cancer: recent advances in molecular diagnosis. AB - Chronic pancreatitis is an inflammatory disease of the pancreas, characterized by a progressive destruction of the exocrine and endocrine pancreas, leading both to exocrine and endocrine insufficiency. In recent years, our knowledge of this disease has improved, an epidemiological link between chronic pancreatitis and pancreatic cancer has been established, and the molecular alterations underlying their pathogenesis have been partly revealed. Nevertheless, the differentiation of chronic inflammation of the pancreas from cancer of the pancreas remains a great challenge. This overview will point out the present knowledge of the molecular pathogenesis of chronic pancreatitis and pancreatic cancer and will focus on the role of molecular markers for differentiating chronic pancreatitis from pancreatic cancer. PMID- 11385250 TI - Pancreatic carcinoma: imaging update 2001. AB - This paper discusses the most recent advances in imaging of pancreatic carcinomas. The specific modalities discussed include helical computed tomography (HCT) with combined CT angiography, magnetic resonance imaging (MRI) and magnetic resonance cholangiopancreatography (MRCP), and positron emission tomography (PET). Endoscopic ultrasound is discussed in other papers. HCT is believed to be the most efficacious modality for initial detection and staging of pancreatic carcinomas. It has an accuracy rate of about 95-97% for the detection of carcinomas and of virtually 100% for staging unresectable carcinomas. The accuracy for CT staging of a resectable tumor is about 80-85%, with small surface metastases to liver and peritoneum being the most often missed findings. MRI and MRCP are best used for evaluation of patients that cannot receive intravenous contrast media for HCT and of patients who have equivocal HCT findings. PET imaging may be useful for detection of extrapancreatic nodal metastases. PMID- 11385251 TI - Treatment of pancreatic cancer: the role of surgery. AB - Pancreatic cancer shows an aggressive growth behavior which results in an extremely poor prognosis. It is presently the 4th to 5th leading cause of cancer related deaths in Western countries with an incidence of 8-10 new cases per 100,000 inhabitants. Since current conservative oncological therapies fail to influence the long-term outcome, curative resection remains the only possibility with a potential for cure. During the past decades, a considerable decrease in postoperative mortality after pancreatic resection and a significant increase in the resection rate have been achieved. Although several types of pancreatic resection have evolved, standard procedures are the classical Whipple resection for cancers of the pancreatic head and left resection for cancers of pancreatic body and tail. Since the pylorus-preserving Whipple resection and extended Whipple resection are still debated as better alternatives to the classical Whipple procedure, large, controlled clinical trials in patients need to be conducted to reach reliable conclusions. However, there is mounting evidence that the pylorus-preserving Whipple procedure offers a better postoperative outcome than the classical Whipple operation without compromising radicality and thereby the long-term prognosis. Despite the progress in surgical treatment of pancreatic cancer, the overall prognosis following resection remains unsatisfactory to date. It is hoped that progress in multimodality treatment and modern therapies, resulting from both clinical and advanced basic research, can improve the prognosis of this malignancy in the near future. PMID- 11385252 TI - Cystic tumors of the pancreas. AB - The discovery of a cystic lesion in the pancreas implies a challenge for the physician. Approximately 10% are cystic tumors, benign to highly malignant, or true cysts, showing all stages of cellular differentiation, from benign to highly malignant tumors. Malignant cystic tumors are rare and comprise only about 1% of all pancreatic malignancies, they are potentially curable. Therefore, correct diagnosis and treatment of these lesions are of great importance. It is usually not possible to separate a pseudocyst from a benign cyst or a cystic tumor, but there are some signs and findings that could be helpful in the clinical decision. The diagnosis of a cystic pancreatic tumor requires different imaging techniques, including ultrasonography, computerized tomography, magnetic resonance imaging, and magnetic resonance cholangiopancreatography, but to distinguish a pseudocyst or a benign cyst from a potentially malignant lesion can be very difficult. The usefulness of blood tests and investigations of cyst fluid can be questionable. Today, surgical treatment of cystic pancreatic tumors can be performed with low morbidity. Therefore, we conclude that an active strategy with resection of cystic tumors of the pancreas should be recommended. PMID- 11385253 TI - Current options for palliative treatment in patients with pancreatic cancer. AB - Palliative treatment is often the only remaining option in the management of pancreatic carcinoma, but its efficacy is poor due to low tumor sensitivity and inadequate treatment protocols. There are several options of palliative treatment with antitumor or supportive intention. Classical end points of palliative treatment are survival, tumor response, and quality of life. A decade ago, palliative chemotherapy consisted mainly of 5-fluorouracil as the standard agent in combination with either other agents and/or radiotherapy. Only the new antineoplastic drug gemcitabine, which was introduced simultaneously with the definition of novel end points of chemotherapy such as clinical benefit, allowed to achieve some progress. However, while gemcitabine monotherapy appeared to be superior to 5-fluorouracil and improved important parameters of quality of life, it could not provide a significant improvement of survival. A novel concept, therefore, is to improve this beneficial cytostatic response in pancreatic carcinoma using a gemcitabine-based protocol by combining it with antineoplastic drugs such as taxanes or platin analogs. This strategy may have the potential to improve the outcome in palliative chemotherapy of pancreatic carcinoma patients with advanced tumor growth or metastases. Best supportive care in pancreatic cancer consists of the treatment of symptoms, such as pain, jaundice, duodenal obstruction, weight loss, exocrine pancreatic insufficiency, and tumor-associated depression. PMID- 11385254 TI - Prevention of pancreatic cancer and strategies for management of familial pancreatic cancer. AB - At the current time, pancreatic cancer remains a difficult and typically fatal disease. A number of case reports and case-control epidemiologic studies have suggested that familial aggregation plays a role in as many as 10% of all pancreatic cancers. During the last several years, genetic alterations responsible for syndromes linked with pancreatic cancer have been identified. These genes include BRCA2, p16, PRSS1, STK11, and various mismatch repair genes. Unfortunately, most kindreds with a familial aggregation cannot be explained by one of these known genetic syndromes. Recent data from the National Familial Pancreas Tumor Registry at Johns Hopkins have estimated the prospective risk of pancreatic cancer among first-degree relatives of pancreatic cancer patients. The risk was estimated by comparing observed new cases of pancreatic cancer to expected numbers. In families where three first-degree relatives had been diagnosed with pancreatic cancer, the risk of another individual developing pancreatic cancer rose to a 57-fold increase over the basal risk. This article reviews the data concerning familial pancreatic cancer. Additionally, this article reviews the data concerning the histological precursors of invasive ductal adenocarcinoma of the pancreas: pancreatic intraepithelial neoplasias. Further, the current Johns Hopkins methodology used to screen for early pancreatic neoplasia in familial pancreatic cancer patients and in patients with familial Peutz-Jeghers syndrome is discussed. In summary, the notable advances in the field of molecular genetics have allowed for a better definition of the genetics of pancreatic cancer. With this knowledge has evolved a better understanding of several high-risk clinical syndromes associated with pancreatic cancer, familial pancreatic cancer, and the evolution of strategies to screen high-risk families for early pancreatic neoplasia. PMID- 11385255 TI - Prognostic parameters determining survival in pancreatic carcinoma and, in particular, after palliative treatment. AB - Prognosis and outcome of patients with pancreatic carcinoma is poor. The aim of the study was to investigate (1) which factors of medical history and clinical status as well as which laboratory parameters determine survival in pancreatic carcinoma and (2) whether specific data can be used as prognostic parameters or for early diagnosis of pancreatic carcinoma. In total, 287 patients with pancreatic carcinoma were enrolled in the study. In 193 subjects, only palliative treatment was possible. Survival was assessed using univariate survival probability curves by Kaplan-Meier. Comparison of patient groups with regard to survival was achieved using the log-rank test. Multivariate analysis was carried out using the Cox regression model. Overall, 22 factors, showing a significant impact on survival in pancreatic carcinoma were found, e.g., tumor-associated factors such as (1) tumor stage according to the UICC classification including TNM-based staging, grading, tumor site, and vascular infiltration; (2) preoperative habits and signs and symptoms (physical condition, pain, loss of appetite, ethanol consumption); (3) change of laboratory parameters (CA 19-9, bilirubin, prothrombin time, urea, C-reactive protein), and (4) type of intervention (surgical approach, R0/1/2 resection). Using multivariate analysis, seven factors (UICC tumor stage and site, surgical intervention including number of resected lymph nodes, chemotherapy, occurence of a carcinoma in relatives, preoperative physical condition, night sweat) were determined. In the 193 patients with palliative treatment, only ten factors (among them UICC tumor stage including the presence of metastases; data from the medical history such as physical condition, loss of appetite, and carcinoma in relatives, and laboratory parameters including prothrombin time, protein content, and aspartate aminotransferase levels) were found to be important. Chemotherapy had the strongest impact on survival which was confirmed by multivariate analysis, followed by tumor stage (UICC) and preoperative appetite. Besides tumor associated determinants, data from the medical history, and pathological laboratory parameters, the prognosis in pancreatic carcinoma is considerably determined by the treatment such as interventional and/or using antineoplastic agents. PMID- 11385256 TI - Acute cerebrovascular disease in women. AB - In 2,000 consecutive stroke patients collected in a prospective hospital-based stroke registry over a 10-year period, we assessed whether stroke in men and women was different in respect to vascular risk factors, clinical features and natural history. The frequency of the different variable in men and women was analyzed by means of univariate analysis and logistic regression models. Women accounted for 48% of the study population (n = 967) and were older than men (mean age 75 vs. 69 years, p < 0.001). In the age group of 85 years or older, stroke was more frequent in women than in men (69.8 vs. 30.2%, p < 0.001). Women showed a higher frequency of cardioembolic infarction and a lower occurrence of lacunar infarction and stroke of undetermined cause than men. In-hospital mortality (17.4 vs. 13.3%) and length of hospital stay (19.6 vs. 16.7 days) was significantly higher (p < 0.001) in women than in men. In the model based on demographic variables and cardiovascular risk factors, obesity, heart failure, atrial fibrillation and age were significant predictors of stroke in women, while intermittent claudication, ischemic heart disease, chronic obstructive pulmonary disease, cigarette smoking and alcohol abuse were predictors in male sex. Hypertension and limb weakness were predictors for stroke in women, and absence of neurological deficit at hospital discharge, lacunar syndrome and ataxia were predictors in men in the models based on all variables. Women differ from men in the distribution of risk factors and stroke subtype, stroke severity and outcome. Differences in stroke pathology and/or differences in functional anatomy or plasticity of the brain between sexes may account for these findings. PMID- 11385257 TI - Tolerability and efficacy of almotriptan in the long-term treatment of migraine. AB - BACKGROUND: Almotriptan is a highly specific 5-HT(1B/1D) receptor agonist, which acts selectively on blood vessels of the brain. Short-term studies have demonstrated that almotriptan provides rapid, effective and reliable relief of migraine attacks, while offering excellent tolerability. PURPOSE: To assess the long-term tolerability and efficacy of oral almotriptan 12.5 mg administered for every migraine attack over a 1-year period. METHODS: A total of 762 patients treated 13,751 attacks (1-97 per patient); 61.5% of attacks were treated with one 12.5-mg dose, while for 38.5% of attacks, patients took a second dose within 24 h. RESULTS: Three hundred and ninety-one patients (51.3%) experienced a total of 1,617 adverse events (AEs). The majority (88.6%) of AEs were of mild-to-moderate intensity, and only 28.8% of AEs were considered to be related to the study drug. Only 2 patients experienced serious AEs possibly related to almotriptan, syncope and chest pain; both recovered without any sequelae. Patients reported at least 1 AE in 11% of attacks treated. The incidence of AEs decreased during the study. Only 6 (0.8%) study withdrawals were due to AEs considered to be related to almotriptan. Tolerability was not compromised in patients taking 2 doses of almotriptan or in those using migraine prophylactics. Patient age or sex did not influence the incidence of AEs. There was no evidence of tachyphylaxis in those patients completing the study. Pain relief at 2 h after the initial dose was achieved in 84.2% of moderate/severe attacks. Patients were pain free at 2 h after dose in 58.2% of all attacks. Older patients (> 40 years) tended to respond better than younger ones (< 40 years). Efficacy was not modified by use of migraine prophylactics or hormonal contraceptives. Efficacy measurements were consistent on treating repeated moderate/severe migraine attacks. CONCLUSION: This large, open study indicates that the new, specific 5-HT(1B/1D) agonist almotriptan, at a dose of 12.5 mg, is a well tolerated and effective treatment for migraine pain when used over a period of up to 1 year. PMID- 11385258 TI - Multiperspective assessment of peripheral nerve involvement in diabetic patients. AB - To assess the relationship between peripheral nerve involvement and the patient's perception of his own quality of life, we studied 36 consecutive out-patients affected by insulin-dependent diabetes mellitus without other diabetic complications other than neuropathy (20 men, 16 women; mean age 39.1 years). We used clinical (Semmes-Weinstein, vibration perception threshold, muscle strength, osteotendinous reflexes), neurophysiological (sural, peroneal and ulnar nerves), metabolic (glycosylated haemoglobin) and patient-oriented (SF-36 and NASS questionnaires) measurements. Patient-oriented physical scores were significantly related to: (1) neurophysiological findings of the inferior limbs; (2) conventional measurements of sensitivity; (3) metabolic assessment. Conversely, patient-oriented mental scores were significantly related only to metabolic assessment. The patient-oriented measure provided an important perspective of the severity of the disease often closely related with the biological parameters and suggested new ways of interpreting conventional biological measurements. In particular, the peripheral nerve picture appeared strictly related with the physical aspects of the patients' quality of life, while the metabolic picture appeared related with both the mental and physical aspects of the quality of life. PMID- 11385259 TI - Polymyography combined with time-locked video recording (video EMG) for presurgical assessment of patients with cervical dystonia. AB - We assessed 26 patients with cervical dystonia, in whom botulinum toxin (BT) injections had failed, before selective peripheral denervation. We decided to base the decision which muscle should be denervated on both clinical information and EMG data and focussed on the following features: activity at onset or during 'dystonic spasms' (according to the concept of the 'leading' dystonic muscle), paradoxical activity during voluntary head movements causing restriction of head movements opposite the side of head turn or tilt and activity when symptoms deteriorated during walking. To identify these muscles we developed a new recording system that integrates simultaneous video-taping and polymyography (video EMG) by means of a digital counter, driven by the recording software (resolution 0.1 s), that was fixed in view of the video camera. This system time locked clinical signs with relevant EMG activity thus allowing demonstration of the above features. These were found in 68% of dystonic muscles with each of them being present in approximately 40%. Video EMG allows an integrated approach to identify overactive neck muscles in patients with cervical dystonia taking into account both relevant clinical findings and EMG data. PMID- 11385260 TI - Drug-induced headache: long-term follow-up of withdrawal therapy and persistence of drug misuse. AB - Patients suffering from migraine, tension-type headache (TTH), or combined headache (CH) are at risk of developing drug-induced headache (DIH) due to regular use of analgesics, ergot alkaloids, and triptans. The aim of our study was to determine (1) the clinical features of DIH, (2) the outcome of withdrawal therapy using high methodological standards, and (3) predictors which could explain the high relapse rate (more than 40%) after a previously successful withdrawal therapy. We retrospectively reviewed 103 patients with migraine or TTH who underwent withdrawal therapy between 1994 and 1998. The long-term follow-up (2-4 years after therapy) was conducted by phone and by specially trained psychologists using a structured interview which enclosed characteristics of headache and medication behavior as well as patients global assessment of success. Complete sets of data were available from 83 patients (38 migraine, 26 TTH, 19 CH). The most frequently abused drugs were caffeine-combined analgesics (24%), followed by caffeine-combined ergotamines (19%), pure ergot alkaloids (17%), and monoanalgesics (17%). 48.5% of the patients suffered an abuse relapse within 4 years and developed the complete features of DIH again. Analgesic and ergot alkaloid combinations with caffeine lead significantly more often to a relapse. A long-term successful therapy is connected with a significant reduction of the frequency of headache attacks. Under relapse conditions, the patients reached their former headache frequency level. The data show a higher relapse rate than previously assumed and that certain substance groups bear a higher relapse risk. PMID- 11385261 TI - Radiculopathy and myelopathy in patients with primary cervical dystonia. AB - In a prospective series of 34 incident patients with primary cervical dystonia (CD), 6 showed clinical or radiological signs of cervical radiculopathy (RP) or myelopathy (MP) during the course of their movement disorder. Age at onset in these patients was in the range reported for pure spondylotic cervical RP without an accompanying movement disorder. Radiologically, spondylosis was mild in 1 case and absent in 2 cases. The intervals between onset of CD and RP were shorter than in literature reports of RP/MP in dystonic-athetotic patients of cerebral paresis. Clinically, RP/MP in patients with CD mostly appears at mid-cervical levels, whereas cases with cerebral paresis are more frequently affected at higher cervical segments. We propose that RP/MP does not occur more frequently in CD than in pure spondylotic cervical RP. PMID- 11385262 TI - Ataxic form of chronic inflammatory demyelinating polyradiculoneuropathy: clinical features and pathological study of the sural nerves. AB - We investigated clinical and pathological features of the sural nerves of 5 patients with the ataxic form of chronic inflammatory demyelinating polyradiculoneuropathy (CIDP) and compared these features with those of chronic ataxic neuropathies due to other causes. The CIDP patients presented with slowly progressive ataxia with deep sensory impairment. The durations of the symptoms from onset were relatively short in CIDP (4-8 months) and cancer (3 and 10 months), but long in chronic idiopathic ataxic neuropathy (24-260 months). Corticosteroid therapy elicited a good response in all the patients with CIDP, but a poor response in the patients with other ataxic neuropathies. Sural nerve biopsy of CIDP patients showed a slight or moderate loss of myelinated fiber. This report suggests that ataxic form of CIDP is a steroid-responsive ataxic neuropathy, and large myelinated fibers of the sural nerves in ataxic form of CIDP were better preserved than those in nerves with other chronic ataxic neuropathies. PMID- 11385263 TI - Brucellar spinal epidural abscess of cervical location: report of four cases. AB - Spinal epidural abscesses account for 1 or 2 of every 10,000 hospital admissions, Staphylococcus aureus being the bacterium most frequently involved. Brucellosis is a disorder of worldwide distribution, relatively frequent in South America and in Mediterranean countries in Europe and Africa. Whilst in the USA only 200 cases are reported every year, in Spain it is the most frequent zoonosis. This systemic disease seldom produces spondylodiscitis which in a minority of cases may be complicated by spinal epidural abscesses, in general of lumbar location. The purpose of this article is to analyse 4 cases of brucellar spinal epidural abscess of cervical location and diagnosed in the Province of Teruel, Spain, an endemic area for the disease, through 10 consecutive years (1990-1999). We consider noteworthy the following facts: the first case was a technical employee who acquired the infection in our laboratory of microbiology, the second presented with an extensive purulent collection invading prevertebral and retropharyngeal regions, the third case was cured only with antibiotics without residual deficits. In the fourth case we were not able to demonstrate spondylodiscitis accompanying the epidural abscess at the C2-C6 levels. We discuss especially the epidemiological aspects of brucellosis, the existence of epidural abscess without spondylodiscitis, the clinical manifestations, the diagnosis by means of magnetic resonance imaging, specific serological tests for Brucella, antibiotic treatment and the prognosis of our cases. PMID- 11385264 TI - Genetic association analysis of alpha-1-antichymotrypsin polymorphism in Parkinson's disease. AB - alpha(1)-Antichymotrypsin (ACT) gene has been suggested as a susceptibility factor for Parkinson's disease (PD) and might be related to the onset of PD. We replicated these findings in a Chinese population. The results demonstrated that the ACT genotypic and allelic distributions showed no significant differences between the PD patient and the control groups. The age at onset was younger in the heterozygotes than in the homozygotes (p = 0.042). We suggest that the ACT polymorphism might play some role in the pathogenesis of PD, especially in the onset. PMID- 11385265 TI - Botulinum toxin antibody testing: comparison between the immunoprecipitation assay and the mouse diaphragm assay. AB - Antibodies against botulinum toxin (BT) are currently best detected by the mouse diaphragm assay (MDA). Nevertheless, the MDA still has disadvantages, so that an immunoprecipitation assay (IPA) was introduced recently. We sought to compare the results of both tests. 33 samples from patients with cervical dystonia and complete or partial BT therapy failure underwent blinded simultaneous IPA and MDA testing. 27 (82%) samples showed concordant results, 17 (52%) being positive and 10 (30%) negative for both IPA and MDA resulting in a significant association of the dichotomous test results (Fisher's exact test, p < 0.01). The other six samples (18%) showed discordant results, all being IPA-negative and MDA-positive. This excess of MDA-positive results was also significant (Sign rank test, p = 0.03). IPA and MDA results showed a very strong and significant qualitative and quantitative association. The IPA seems to be less sensitive than the MDA for detection of low BT-AB titres, but the clinical relevance of this still needs to be established. Since the IPA is simpler, faster and cheaper than the MDA and avoids sacrifice of animals, it could become the preferred BT-AB test. PMID- 11385266 TI - Discrepancy between dysmetric centrifugal movements and normometric centripetal movements in psychogenic ataxia. AB - A method to unravel an aberrant motor behaviour in psychogenic ataxia is reported. The kinematic features of fast reaching movements in the vertical plane are described in a patient presenting a psychogenic ataxia. The procedure compared centrifugal and centripetal movements. Path ratios were computed for each phase, as well as the ratios of the paths for centrifugal and centripetal directions. Trajectories of centrifugal phases were erratic but centripetal movements were very regular, whereas both centripetal and centrifugal movements were irregular in patients presenting an organic cerebellar syndrome. A similar incongruity between movements in opposite directions is also shown for a second patient exhibiting psychogenic ataxia. Discrepancies between the centrifugal phase and the centripetal phase of multi-joint reaching movements support the diagnosis of a psychogenic movement disorder. PMID- 11385267 TI - Time of developing motor deficits before radiotherapy as a new and relevant prognostic factor in metastatic spinal cord compression: final results of a retrospective analysis. AB - The aim of this retrospective analysis was to determine the prognostic value of the time of developing motor deficits before radiotherapy (RT) and to confirm our preliminary results by achieving statistical significance. Of 529 patients receiving spinal irradiation (1994-1998) 131 fulfilled the selection criteria. Three groups were formed according to time of developing motor deficits: 1-7 days (n = 51), 8-14 days (40), >14 days (40). Motor function was graded before, 2 weeks and 3 months after RT. After 2 weeks, patients developing motor deficits for >14 days showed improvement more often than patients of the other groups (65 versus 32.5 and 4%, p < 0.001). After 3 months results were comparable (p < 0.001). A slower development of motor deficits predicts a better functional outcome. Time of developing motor deficits before RT is a relevant prognostic factor in metastatic spinal cord compression. PMID- 11385268 TI - Plasmapheresis in the treatment of ataxic sensory neuropathy associated with Sjogren's syndrome. AB - Sjogren's syndrome (SS) is an important but poorly recognized cause of peripheral neuropathy. Several forms of peripheral nerve dysfunction occur, including trigeminal sensory neuropathy, mononeuropathy multiplex, distal sensorimotor polyneuropathy and pure sensory neuronopathy. The pathological findings vary and the definite treatment is not known. Here we present 4 cases of acute ataxic sensory polyneuropathy with SS, and the experience of treatment with plasmapheresis (PP). The 4 patients were all females; ages ranged from 30 to 58 years. All had prominent loss of kinesthetic and proprioceptive sensation. The course ranged from acute to subacute onset. Patients were treated with 5-9 sessions of PP. Two patients with initiation of treatment within 2 weeks of onset showed dramatic and sustained responses after PP, while the other 2 had no detectable effects. Our experience showed that PP should be considered in patients who present with sensory neuropathy associated with SS, and the treatment should be given as early as possible. PMID- 11385269 TI - Comparison of preference for rizatriptan 10-mg wafer versus sumatriptan 50-mg tablet in migraine. AB - Rizatriptan (MAXALT, a registered trademark of Merck & Co. Inc.) is a selective 5 HT(1B/1D) receptor agonist with rapid oral absorption and early onset of action in the acute treatment of migraine. This randomized, open-label, crossover outpatient study assessed the preference of 481 patients for rizatriptan 10-mg rapidly disintegrating tablets versus sumatriptan (IMIGRAN, a registered trademark of GlaxoWellcome PLC) 50-mg tablets in the treatment of a single migraine attack with each therapy. Almost twice as many patients preferred rizatriptan 10-mg rapidly disintegrating tablet to sumatriptan 50-mg tablet (64.3 vs. 35.7%, p < or = 0.001). Faster relief of headache pain was the most important reason for the preference, cited by 46.9% of patients preferring rizatriptan and 43.4% of patients who preferred sumatriptan. Headache relief at 2 h was 75.9% with rizatriptan and 66.6% with sumatriptan (p < or = 0.001), with rizatriptan being superior to sumatriptan within 30 min of dosing. Fifty-five percent of patients were pain free 2 h after rizatriptan, compared with 42.1% treated with sumatriptan (p < or = 0.001), rizatriptan being superior within 1 h of treatment. Forty-one percent of patients taking rizatriptan were pain free at 2 h and had no recurrence or need for additional medication, compared to 32.3% of patients on sumatriptan. Rizatriptan was also superior to sumatriptan in terms of the proportions of patients with no nausea, phonophobia or photophobia, and patients with normal function 2 h after treatment intake (p < 0.05). More patients were (completely, very or somewhat) satisfied 2 h after treatment with rizatriptan (73.3%) than 2 h after treatment with sumatriptan (59.0%) (p < or = 0.001). Additionally, 2 h after the dose, more patients found rizatriptan to be very convenient, convenient or somewhat convenient (87.2%) than they did sumatriptan (76.3%) (p < or = 0.001). Both active treatments were well tolerated. The most common side effects with rizatriptan and sumatriptan were nausea (6.6 and 6.9% of patients, respectively), dizziness (6.1 and 5.8%) and somnolence (7.4 and 6.7%). PMID- 11385270 TI - Successful thrombolytic treatment of intracranial carotid occlusion due to dissection. PMID- 11385271 TI - Atypical paraneoplastic brainstem encephalitis associated with anti-ri-antibodies due to thymic carcinoma with possible clinical response to immunoglobulins. PMID- 11385272 TI - Botulinum toxin for treatment of jaw opening dystonia in Hallervorden-Spatz syndrome. PMID- 11385273 TI - Neurosarcoidosis associated with interferon therapy. PMID- 11385274 TI - P-glycoprotein, multidrug-resistance-associated protein and Parkinson's disease. PMID- 11385275 TI - Isolated lesions limited to the bilateral substantia nigra on MRI associated with influenza A infection. PMID- 11385276 TI - Avellis' syndrome: the neurological-topographical correlation. PMID- 11385277 TI - Acute isolated uvular infarction. PMID- 11385278 TI - Evidence of the significance of secondary excitations of the vocal tract for vocal intensity. AB - In the production of voiced speech the main excitation of the vocal tract occurs during the glottal closing phase when the rate of change of the flow reaches its absolute maximum. The level of this maximum, the negative peak amplitude of the differentiated glottal flow, correlates strongly with sound pressure level. This study presents a straightforward method that yields a numerical value to characterize the effect of the main excitation on vocal intensity. The method, energy ratio by modified excitation (ERME), utilizes the glottal flow and the vocal tract transfer function estimated by inverse filtering and it synthesizes two signals based on the linear source-filter theory. The first sound is produced using the glottal flow given by inverse filtering per se. The second signal is synthesized by removing the main excitation from the differentiated flow. ERME is defined as the ratio between the energy of the first synthesized signal and the energy of the second one. It is shown that when the loudness of speech increases, the value of ERME first rises but in the case of very loud voices it starts to decrease. Hence, ERME shows that effects of secondary excitations of the vocal tract that occur during glottal opening become important in the production of very loud voices. PMID- 11385279 TI - The influence of acoustic and perceptual factors on perceived hypernasality in the vowel. AB - The purpose of this study was to examine the influence of acoustic and perceptual factors of speech on listeners' perceived hypernasality in the vowel [i]. The isolated syllable [pi] produced by 22 children with hypernasal speech and 6 noncleft children was rated by 10 listeners. These speech samples were then divided into two groups: (1) the samples (n = 14) that received inconsistent ratings from each listener or variable ratings among listeners (i.e., unreliable ratings) and (2) the samples (n = 14) that received consistent ratings from each listener and similar ratings among listeners (i.e., reliable ratings). These results suggest that the severity of hypernasality was easy to rate in some speech samples and not in others. Voice quality deviation and a particular type of spectral change that related to the severity of hypernasality could be factors that influence perceived hypernasality. PMID- 11385280 TI - Speech patterns in children with velo-cardio-facial syndrome. Two case studies. AB - The purpose of this study was to document the articulation patterns in children with velo-cardio-facial syndrome. The subjects were 2 children with this syndrome, aged 6 years 6 months and 3 years 11 months. Phonetically transcribed speech samples collected by means of a picture naming test formed the basis for this study. The speech samples were subjected to two types of analyses: independent analyses describing the children's sound productions regardless of their relation to the adult targets and relational analyses comparing the children's productions with the adult standard form. It was found that the articulatory problems in velo-cardio-facial syndrome are not simply due to a delay in speech sound acquisition. Articulation errors found in this syndrome are uncharacteristic of normal speech development. Whether errors are syndrome specific awaits further investigation. PMID- 11385281 TI - The effects of changes in speaking rate on nasal airflow and the perception of nasality. AB - The effects of variation in speaking rate on relative nasal airflow (percent nasal flow) and on the perception of nasality were examined. In addition, the effects of gender and speech rate elicitation techniques (metronome-controlled, self-controlled) were examined. Nineteen normal speakers each produced a stimulus phrase containing nonnasal sounds. Oral and nasal airflows were measured using the Rothenberg aerodynamic system. Results indicated that percent nasal flow and perception of nasality were both greater at slow speaking rates compared to normal and fast rates. Males were perceived as more nasal than females. The metronome-controlled rates were associated with greater nasality than the self controlled rates. Discussion focuses on physiological correlates to these findings. PMID- 11385282 TI - Human mast cell transcriptome project. AB - After draft reading of the human genome sequence, systemic analysis of the transcriptome (the whole transcripts present in a cell) is progressing especially in commonly available cell types. Until recently, human mast cells were not commonly available. We have succeeded to generate a substantial number of human mast cells from umbilical cord blood and from adult peripheral blood progenitors. Then, we have examined messenger RNA selectively transcribed in these mast cells using high-density oligonucleotide probe arrays. Many unexpected but important transcripts were selectively expressed in human mast cells. We discuss the results obtained from transcriptome screening by introducing our data regarding mast-cell-specific genes. PMID- 11385284 TI - Dual TCR-expressing T lymphocytes in health and disease. AB - The authors briefly review recent experimental advances in elucidating the role of dual T cell receptor (TCR)-expressing lymphocytes in the development of diseases with special emphasis on autoimmunity. Moreover, they summarize present knowledge about these cells concerning their proportion among peripheral blood mononuclear cells, their functionality, and their impact on allorecognition and memory both in humans and in mice. Finally, they describe disease-associated clonal expansions of dual TCR-expressing cells in humans, most of which have been observed in peripheral T cell malignancies. Other cases occurred in inflammatory bowel disease and in HIV infection. They propose that expression of two distinct TCR on malignant T lymphocytes might be much higher than is suggested by the few cases described so far, and that their presence might impinge on therapeutic immunization strategies which make use of the TCR itself as a target. PMID- 11385283 TI - Fc epsilon ri-mediated activation of transcription factors in antigen-presenting cells. AB - Professional antigen-presenting cells (APC) such as monocytes and dendritic cells (DC) bearing high-affinity IgE receptors (Fc epsilon RI) efficiently present IgE bound antigens to T cells. Fc epsilon RI expression is upregulated on APC from atopic donors, especially in inflamed tissues. These data suggest a pathophysiological concept of an IgE-mediated delayed-type hypersensitivity reaction in atopic diseases. However, Fc epsilon RI ligation also leads to the synthesis of proinflammatory cytokines and other molecules involved in inflammatory reactions. The investigation of transcription factors mediating these effects has only recently commenced. In general, members of the NF-kappa B family are known to regulate APC function and differentiation, with the RelB subunit being especially important in DC generation. In addition, Ikaros and PU.1 have also been shown to be essential factors for DC differentiation, whereas Oct 2 is upregulated by differentiation towards macrophages. Recently, Fc epsilon RI has been demonstrated to induce NF-kappa B activation via I kappa B-alpha serine phosphorylation and degradation in monocytes and DC. Inhibitors of NF-kappa B activation such as N-acetylcysteine or N-tosyl-L-phenylalanine chloromethyl ketone can suppress Fc epsilon RI-induced TNF-alpha and MCP-1 release. Interestingly, in human epidermal Langerhans' cells (LC), NF-kappa B activation can only be observed when large amounts of Fc epsilon RI are present. In addition, the composition of NF-kappa B complexes differs between monocytes, monocyte-derived DC and LC, suggesting a cell type-specific regulation. Moreover, the transcription factor NFAT is induced upon Fc epsilon RI ligation in human APC. The elucidation of further transcription factors involved in Fc epsilon RI signaling in APC should contribute to the employment of new inhibition strategies for the treatment of atopic and other inflammatory diseases. PMID- 11385285 TI - Does IL-4 play a role in the expansion of V beta 8a T cell receptor-bearing cells? AB - BACKGROUND: Peripheral blood mononuclear cells (PBMC) of subjects allergic to the insect-derived allergen Chi t 1--9 are characterized by an allergen-induced pronounced proliferation and increased expression of activation markers (CD25, HLA-DR, CD23). T cell lines showed an elevated percentage of V beta 8a-positive cells following stimulation by Chi t 1--9. OBJECTIVE: The aim of the present study was to investigate whether V beta 8a dominance plays an important role in PBMC short-term cultures (24 h) as well. The role of exogenous added cytokines, especially IL-4, has been determined. METHODS: The T cell receptor repertoire was measured with 16 monoclonal antibodies to epitopes on the variable region of the beta chain by flow cytometry. Patients allergic to Chi t 1--9 were compared to nonallergic subjects as well as to subjects with other occupational allergies. In addition, cytokines were determined intracellulary by flow cytometry. Studies were performed with PBMC cultured for 24 h. RESULTS: After cultivation for 24 h without or with different stimuli (cytokines, allergen, phytohaemagglutinin), changes in the T cell receptor profile and the cytokine profile were measurable compared to the baseline value (without cultivation). Stimulation with IL-4 revealed increased percentages of V beta 8a-expressing cells in Chi t 1--9 sensitized patients. This IL-4-induced V beta 8a increase did not occur in PBMC from the two control subject groups (non-allergic and allergic to other allergens than Chi t 1--9). CONCLUSION In conclusion, the dominance of certain T cell receptor types seems to arise due to the exposure to specific allergens and cytokine production. Some T cell receptors are often affected, for example V beta 8a, whereas others only show minor variations. V beta 8a expression obviously plays an important role in Chi t 1-9 allergy. PMID- 11385286 TI - High-level expression in mammalian cells of recombinant house dust mite allergen ProDer p 1 with optimized codon usage. AB - BACKGROUND: The major house dust mite allergen Der p 1 is associated with allergic diseases such as asthma. Production of recombinant Der p 1 was previously attempted, but with limited success. The present study describes the expression of recombinant (rec) ProDer p 1, a recombinant precursor form of Der p 1, in CHO cells. METHODS: As optimization of the codon usage may allow successful overexpression of protein in mammalian cells, a synthetic gene encoding ProDer p 1 was designed on the basis of the codon usage frequently found in highly expressed human genes. Gene synthesis was accomplished from a set of 14 mutually priming overlapping oligonucleotides and after two runs of polymerase chain reaction. RESULTS: COS cells transiently transfected with the synthetic ProDer p 1 gene produced up to 5--10 times as much ProDer p 1 compared with the expression level obtained after transfection with the authentic gene. To stably express the recombinant allergen, CHO-K1 cells were transfected with the ProDer p 1 synthetic gene, and one amplified recombinant clone produced up to 30 mg of recProDer p 1 per liter in the culture medium before purification. recProDer p 1 was secreted as an enzymatically inactive single-chain molecule presenting three glycosylated immunoreactive forms of 41, 38 and 36 kD. When examined with respect to direct binding, recProDer p 1 and natural Der p 1 displayed very similar IgE reactivities. However, IgE inhibition and histamine release assays showed a much higher reactivity to natural Der p 1 compared to recProDer p 1. CONCLUSIONS: These data indicated that codon optimization represents an attractive strategy for high-level production of allergen in mammalian cells. PMID- 11385287 TI - Identification of cyclophilin as an IgE-binding protein from carrots. AB - BACKGROUND: Plant food allergies have been associated with pollenosis, although most of the causative allergens are as yet undefined. It is important to elucidate the properties of plant food allergens in order to minimize a patient's risks in food selection. The purpose of the present study was to examine and characterize the IgE-binding proteins in carrots as possible allergens by using patients' sera. METHOD: IgE-binding proteins in carrot extract were screened by an immunoblot technique using sera of patients with atopic dermatitis (selected based upon a case history of food allergies). An allergenic protein was purified from carrot extract by chromatographic procedures. The N-terminal amino acid sequence of allergenic protein was determined and subjected to a computer homology search. Cross-reactivity between carrot and birch allergens was examined by immunoblotting. RESULTS AND CONCLUSION: A unique, approximately 20-kDa allergenic protein that reacted with about 14% of patients' sera was isolated and characterized. The N-terminal amino acid sequence of the purified protein was found to be homologous with those of plant cyclophilins. This allergen exhibited a peptidyl-prolyl cistrans isomerase activity, which was inhibited by the conjugation of cyclosporin A. These properties of the allergenic protein isolated from carrot identified it as a cyclophilin, a possible plant food allergen. No cross-reactivity between this 20-kDa carrot allergen and Bet v 7, a birch pollen cylcophilin, was observed. PMID- 11385288 TI - Identification of allergen fractions of wheat flour responsible for anaphylactic reactions to wheat products in infants and young children. AB - Wheat is a food allergen which occasionally causes anaphylactic reactions exclusively in young children. There is very little knowledge of the clinical outcome in cases of food-related anaphylaxis and the differences in the allergenic protein components of food involved, comparing individuals who have suffered from an anaphylactic reaction with other individuals. The objectives of the present study were to examine the clinical features of 7 young children who had experienced anaphylactic reactions after ingesting wheat flour-containing products, and to analyze the allergens in wheat flour responsible for the anaphylactic symptoms. We measured the total IgE levels and the levels of IgE antibodies specific to wheat flour and performed IgE immunoblotting, comparing the sera from these children with sera from patients with atopic dermatitis. All sera from children who had experienced anaphylactic reactions were found to be positive for IgE specific to wheat. IgE immunoblotting revealed that 3 of these 7 children had sera showing reactivity to components of the salt-soluble protein fraction (16, 35--67 and 94 kD) and salt-insoluble protein-containing fraction (16, 38 and 70 kD) and 4 had no sera showing reactivity to components of the salt soluble fraction. Patients with atopic dermatitis showed similar staining patterns. Various proteins in wheat flour could be allergens responsible for anaphylaxis and atopic dermatitis in infants or young children. Our findings suggest that these two clinically diverse allergic diseases do not necessarily represent responses to different allergenic proteins of wheat. PMID- 11385289 TI - Multiple pollen sensitization: a molecular approach to the diagnosis. AB - BACKGROUND: Sensitization to multiple pollen species is a frequent diagnostic event. Several allergenic molecules with a high level of homology have been identified in divergent pollen families and named panallergens. METHODS: We sought to define the criteria to evaluate the prevalence of the multiple pollen sensitization, to identify specific markers of this condition, and to correlate them with the underlying allergic disease. Patients presenting an allergic respiratory disease underwent skin testing with 23 pollens. Patients fulfilling predefined selection criteria were grouped and classified as having multiple pollen sensitization. Patients in each subgroup were tested for IgE to rBet v 2, rJun o 2, rBet v 1, rPhl p 5 and bromelain. Demographical, allergological and clinical data were recorded in the subgroup of patients with multiple pollen sensitization. RESULTS: Seventeen percent of the pollen-sensitized patients formed the multiple pollen-sensitized subgroup. These subjects were positive for most of the pollen species tested regardless of known exposure to them. None of the subjects sensitized to less than six pollen species were positive to panallergens, whereas 55% of the sera of the multiple pollen-sensitized group were positive to rBet v 2, and 15% to rJun o 2. IgE to rBet v 1 and rPhl p 5 were found positive in all the subgroups. Age, gender, bronchial asthma, oral allergy syndrome, skin test reactivity and previous specific immunotherapy differed significantly when these two subsets were considered. CONCLUSIONS: Allergy diagnosis based on allergenic molecules is crucial in the patient with multiple pollen sensitization. This condition appears to be determined by the sensitization to defined allergenic components (panallergens) rather than by pollen of multiple species as such. Detection of IgE to nonpanallergenic molecules allows to identify more relevant allergenic sources. Clinical aspects of the underlying allergic disease (e.g. asthma and oral allergy syndrome) seem to be differently related to IgE reactivity to panallergens. PMID- 11385291 TI - Controlled comparison of the efficacy and safety of cetirizine 10 mg o.d. and fexofenadine 120 mg o.d. in reducing symptoms of seasonal allergic rhinitis. AB - BACKGROUND: The efficacy, onset and duration of action and safety of cetirizine 10 mg o.d., fexofenadine 120 mg o.d., and placebo were compared in this investigator-blinded, crossover study involving the use of the Vienna Challenge Chamber. METHODS: 40 volunteers with seasonal allergic rhinitis were exposed to a controlled grass pollen concentration for 6 h on 2 consecutive days. Subjective symptoms and objective measurements were recorded during the allergen exposure periods. RESULTS: Both active medications were significantly more effective than placebo and had a comparable onset of action in alleviating the symptoms of seasonal allergic rhinitis. The efficacy of both active drugs was comparable for the first 4 h after administration of the drugs on day 1 and day 2. However from 22 to 24 h after the first dose cetirizine was significantly superior to fexofenadine for the major symptom complex score and for sneezing. Concerning the total symptom complex score at day 2 fexofenadine could not reach superiority to placebo. No serious adverse events were reported. CONCLUSIONS: Cetirizine and fexofenadine were significantly better than placebo, also in reducing the symptom of nasal congestion. However cetirizine appears to have a longer duration of action than fexofenadine. PMID- 11385290 TI - Helicobacter pylori infection affects eosinophilic cationic protein in the gastric juice of patients with idiopathic chronic urticaria. AB - BACKGROUND: Helicobacter pylori, the main cause of gastritis and peptic ulcer, has been associated with idiopathic chronic urticaria (ICU), an immunological skin disorder of unknown origin. Eosinophilic cationic protein (ECP) is a cytotoxic molecule secreted by the activated eosinophils involved in the pathogenesis of ICU. We assessed serum/gastric juice ECP levels and gastric mucosal eosinophil infiltration in ICU patients infected or not with H. pylori and evaluated the modification after bacterium eradication. METHODS: 33 patients with ICU and 25 dyspeptic controls underwent upper gastrointestinal endoscopy for histological evaluation and assessment of H. pylori infection. One-week triple therapy was given to H. pylori-positive patients. Serum and gastric juice ECP levels, eosinophil infiltration from gastric mucosal sections and urticaria symptoms were evaluated in all patients at enrollment and 8 weeks after eradication. RESULTS: 19 of 33 (57%) ICU patients and 16 of 25 (64%) controls were found to be infected with H. pylori. Serum ECP was significantly higher in ICU patients compared to controls, regardless of infectious status. Gastric juice ECP and gastric eosinophil infiltration were significantly higher in infected ICU patients when compared both to uninfected ICU patients and controls. H. pylori eradication determined a significant decrease in gastric juice ECP and gastric eosinophil infiltration only in ICU patients. Moreover, a total or partial remission of urticaria symptoms was observed only in ICU patients in whom the bacterium was eradicated. CONCLUSIONS: Although H. pylori infection affects gastric juice ECP and eosinophil infiltration of ICU patients, the role of the bacterium in the pathogenesis of this skin disorder still remains uncertain. PMID- 11385292 TI - Lymphocyte diversity in a 9-year-old boy with idiopathic CD4+ T cell lymphocytopenia. AB - Since CD4+ lymphocytopenia can be caused by disturbed thymic T-cell maturation, we investigated the T-cell subsets of a 9-year-old boy fulfilling the diagnostic criteria for CD4+ lymphocytopenia in a follow-up period of 4 years. We found (I) reduced CD45RA expression, (II) enhanced CD45RO expression and (III) a significant increase in gamma delta TCR-bearing T cells. An accelerated apoptosis (11%) was observed in the CD45RO+, but not CD45RA+ subset. These findings provide evidence that a disturbed thymic T-cell maturation process might play a role in the pathogenesis of CD4+ lymphocytopenia. PMID- 11385293 TI - Anaphylaxis due to hypersensitivity to Anisakis simplex. AB - Anaphylactic reactions are among the most frequent causes for requests for emergency medical care. It is possible that no clear precipitating factor can be found delaying the onset of adequate treatment and increasing the risk of new episodes of anaphylaxis in the future. Hypersensitivity to Anisakis simplex, an intestinal parasite found in fish, is an unusual cause of anaphylaxis but it should always be borne in mind in countries where a great deal of fish is consumed, especially if it is eaten raw or undercooked. 3 cases of anaphylaxis due to hypersensitivity to A. simplex are described. PMID- 11385294 TI - Effects of Betel chewing on the central and autonomic nervous systems. AB - Betel chewing has been claimed to produce a sense of well-being, euphoria, heightened alertness, sweating, salivation, a hot sensation in the body and increased capacity to work. Betel chewing also leads to habituation, addiction and withdrawal. However, the mechanisms underlying these effects remain poorly understood. Arecoline, the major alkaloid of Areca nut, has been extensively studied, and several effects of betel chewing are thought to be related to the actions of this parasympathomimetic constituent. However, betel chewing may produce complex reactions and interactions. In the presence of lime, arecoline and guvacoline in Areca nut are hydrolyzed into arecaidine and guvacine, respectively, which are strong inhibitors of GABA uptake. Piper betle flower or leaf contains aromatic phenolic compounds which have been found to stimulate the release of catecholamines in vitro. Thus, betel chewing may affect parasympathetic, GABAnergic and sympathetic functions. Betel chewing produces an increase in heart rate, blood pressure, sweating and body temperature. In addition, EEG shows widespread cortical desynchronization indicating a state of arousal. In autonomic function tests, both the sympathetic skin response and RR interval variation are affected. Betel chewing also increases plasma concentrations of norepinephrine and epinephrine. These results suggest that betel chewing mainly affects the central and autonomic nervous systems. Future studies should investigate both the acute and chronic effects of betel chewing. Such studies may further elucidate the psychoactive mechanisms responsible for the undiminished popularity of betel chewing since antiquity. PMID- 11385295 TI - Vaccine- and hepatitis B immune globulin-induced escape mutations of hepatitis B virus surface antigen. AB - Hepatitis B virus surface antigen (HBsAg) vaccination has been shown to be effective in preventing hepatitis B virus (HBV) infection. The protection is based on the induction of anti-HBs antibodies against a major cluster of antigenic epitopes of HBsAg, defined as the 'a' determinant region of small HBsAg. Prophylaxis of recurrent HBV infection in patients who have undergone liver transplantation for hepatitis B-related end-stage liver disease is achieved by the administration of hepatitis B immune globulins (HBIg) derived from HBsAg vaccinated subjects. The anti-HBs-mediated immune pressure on HBV, however, seems to go along with the emergence and/or selection of immune escape HBV mutants that enable viral persistence in spite of adequate antibody titers. These HBsAg escape mutants harbor single or double point mutations that may significantly alter the immunological characteristics of HBsAg. Most escape mutations that influence HBsAg recognition by anti-HBs antibodies are located in the second 'a' determinant loop. Notably, HBsAg with an arginine replacement for glycine at amino acid 145 is considered the quintessential immune escape mutant because it has been isolated consistently in clinical samples of HBIg-treated individuals and vaccinated infants of chronically infected mothers. Direct binding studies with monoclonal antibodies demonstrated a more dramatic impact of this mutation on anti-HBs antibody recognition, compared with other point mutations in this antigenic domain. The clinical and epidemiological significance of these emerging HBsAg mutants will be a matter of research for years to come, especially as data available so far document that these mutants are viable and infectious strains. Strategies for vaccination programs and posttransplantation prophylaxis of recurrent hepatitis need to be developed that may prevent immune escape mutant HBV from spreading and to prevent these strains from becoming dominant during the next decennia. PMID- 11385297 TI - Effect of a meal feeding schedule on hepatic glycogen synthesis and gluconeogenesis in rats. AB - We investigated the effect of a meal feeding schedule (MFS) on food intake, hepatic glycogen synthesis, hepatic capacity to produce glucose and glycemia in rats. The MFS comprised free access to food for a 2-hour period daily at a fixed mealtime (8.00-10.00 a.m.) for 13 days. The control group was composed of rats with free access to food from day 1 to 12, which were then starved for 22 h, refed with a single meal at 8.00-10.00 a.m. and starved again for another 22 h. All experiments were performed at the meal time (i.e. 8.00 a.m.). The MFS group exhibited increased food intake and higher glycogen synthase activity. Since gluconeogenesis from L-glutamine or L-alanine was not affected by MFS, we conclude that the increased food intake and higher glycogen synthase activity contributed to the better glucose maintenance showed by MFS rats at the fixed meal time. PMID- 11385296 TI - Exercise training activates large-conductance calcium-activated K(+) channels and enhances nitric oxide production in rat mesenteric artery and thoracic aorta. AB - Exercise training has reversible beneficial effects on cardiovascular diseases, e.g. hypertension, which may result from a decrease in systemic vascular resistance. The purpose of this study was to investigate possible mechanisms associated with the changes in vascular reactivity in large and small arteries with vasoconstrictors and vasodilators in rats after exercise. Wistar-Kyoto rats were trained for 8 weeks (Ex group) on a treadmill and compared with sedentary counterparts (Sed group). After the measurement of blood pressure and heart rate at 8 weeks, rat mesenteric arteries and thoracic aortas were excised and prepared as rings for this study. In addition, special care was taken not to damage the endothelium of the preparations. Our results showed that exercise training for 8 weeks (1) not only prevented an increase in blood pressure but also caused a fall in heart rate, (2) attenuated the contractions induced by both prostaglandin F(2alpha) (PGF(2alpha)) and high K(+) in the mesenteric artery, but reduced the PGF(2alpha)-induced contraction in the aorta only, (3) enhanced the relaxation elicited by acetylcholine (ACh) in both mesenteric arteries and aortas, and (4) increased nitrate [an indicator of nitric oxide (NO) formation] in plasma. The enhancement of ACh-induced relaxation in the mesenteric arteries in the Ex group was suppressed by pretreatment with N(omega) -nitro-L-arginine methyl ester (L NAME), tetraethylammonium (TEA; a nonselective inhibitor of K(+) channels) or charybdotoxin [CTX; a selective inhibitor of large-conductance calcium-activated K(+) (BK(Ca)) channels], whereas in the aorta that response was attenuated by TEA or CTX and almost completely abolished by L-NAME. However, with a combination of L-NAME plus CTX in the mesenteric artery, ACh-induced relaxation was completely abolished in the Sed group, but not in the Ex group. These results suggest that in addition to NO, activation of BK(Ca) channels in the vascular beds, at least in part, also contributes to vasodilatation in animals with exercise training. PMID- 11385298 TI - Expression of human Fas ligand on mouse beta islet cells does not induce insulitis but is insufficient to confer immune privilege for islet grafts. AB - Fas (CD95) and Fas ligand (FasL/CD95L) are involved in programmed cell death and the regulation of host immune responses. FasL has been shown to provide immune privilege, thus prolonging the survival of unmatched grafts in a variety of tissues, such as eyes and testis. In murine FasL (mFasL) transgenic mice, FasL provoked granulocyte infiltration and insulitis in the pancreas. We intended to study whether the expression of human FasL, instead of mFasL, on mouse beta islet cells could avoid granulocyte infiltration, and whether islet cells transgenic for FasL could be used in islet transplantation. We produced transgenic mice in which the human FasL transgene was driven by rat insulin promoter and was expressed exclusively in the pancreas islet cells in ICR mice. In contrast to mFasL transgenic mice, histochemical staining showed that the pancreas was intact in human FasL transgenic ICR mice. However, when human FasL transgenic islet cells were transplanted into allogeneic mice with streptozotocin-induced diabetes, human FasL appeared not to prolong graft survival. Intensive granulocyte infiltration into the islet grafts was observed in recipients (Balb/c mice) which received islet grafts from human FasL transgenic mice, but not from nontransgenic, allogeneic ICR mice on day 31. Our observations suggest that FasL alone is insufficient to confer immune protection, and that other environmental factors might contribute to the formation of immune privilege sites in vivo PMID- 11385299 TI - Utilization of Drosophila eye to probe the functions of two mammalian serine/threonine kinases, Snk and HsHPK. AB - Here we report a quick functional analysis of two mammalian serine/threonine kinases, a serum inducible kinase (Snk) and Homo sapiens hepatoma protein kinase (HsHPK), using Drosophila eye as a model system. We generated transgenic fly lines carrying constructs of both kinases under control of the GAL upstream activating sequence (UAS). Each UAS line was then crossed to a line in which GAL4 expression was driven by one of the following promoters, eyeless (ey), glass or decapentaplegic. Thus, different kinase mutants can be ectopically expressed in a promoter-dependent manner. We observed that the ectopic expression of either the wild-type or active form of Snk driven by the glass promoter resulted in a rough eye phenotype. Nevertheless, the ectopic expression of HsHPK under the control of the ey promoter resulted in a small-eye phenotype. The results of this study demonstrated that ectopic expression of these two mammalian genes could be achieved by the regulation of Drosophila promoters. In addition, the effects of these ectopically expressed genes on eye development could be an implication of their functions with respect to cell proliferation and differentiation. Thus, Drosophila eye, with the powerful genetic tools and vast information on eye development available, can be a useful system to probe the functions of mammalian genes in the postgenome era. PMID- 11385300 TI - Purification of Moloney murine leukemia virus chromatin from infected cells by an affinity method. AB - Our goal was to develop a system to study proteins that associate in vivo with the Moloney murine leukemia virus (M-MuLV) enhancer elements by the isolation of intact proviral chromatin. The M-MuLV long terminal repeats (LTRs) contain tandemly repeated transcriptional enhancer sequences consisting of smaller motifs that bind cellular DNA-binding proteins implicated in transcriptional regulation. The M-MuLV enhancers are also important for disease specificity and latency of disease induction. To enrich for proviral chromatin containing M-MuLV LTR sequences, an affinity purification scheme was employed that relies on the affinity of bacterial Lac repressor protein for Lac operator (LacO) DNA sequences. An infectious M-MuLV recombinant was constructed that contains bacterial LacO sequences inserted into a nonessential region downstream from the 5' LTR of the virus (M-MuLV-LacO). Nuclei from M-MuLV-LacO-infected cells were digested with PvuII (which will liberate an LTR fragment containing LacO sequences), and digested chromatin was leached from the nuclei in hypotonic buffer. M-MuLV-LacO chromatin was then recovered by binding to an affinity matrix consisting of a beta-galactosidase-Lac repressor fusion protein anchored to acrylamide beads by an anti-beta-galactosidase monoclonal antibody [7]. Specifically bound chromatin was eluted under physiological conditions by incubation with the galactose analog isopropyl-beta-D-thiogalactopyranoside. Southern blot analysis confirmed the specific enrichment of M-MuLV proviral chromatin by this method. PMID- 11385302 TI - Sexuality of the elderly. AB - Partial androgen deficiency of the aging male is associated with symptoms collectively accepted as the andropause syndrome. The underlying hormonal changes, the definition of age-dependent cofactors for changing sexuality, and the data on decreasing erectile function are the main topics of this critical analysis. Alterations in libido, ejaculation and sperm quality also have to be considered in order to define a change in male sexuality as part of the natural process of aging. PMID- 11385301 TI - Identification of a potential HIV-induced source of bystander-mediated apoptosis in T cells: upregulation of trail in primary human macrophages by HIV-1 tat. AB - The induction of apoptosis in T cells by bystander cells has been repeatedly implicated as a mechanism contributing to the T cell depletion seen in HIV infection. It has been shown that apoptosis could be induced in T cells from asymptomatic HIV-infected individuals in a Fas-independent, TNF-related apoptosis inducing ligand (TRAIL)-dependent manner if the cells were pretreated with anti CD3. It has also been shown that T cells from HIV-infected patients were even more sensitive to TRAIL induction of apoptosis than they were to Fas induction. Recently, it has been reported that in an HIV-1 SCID-Hu model, the vast majority of the T cell apoptosis is not associated with p24 and is therefore produced by bystander effects. Furthermore, few apoptotic cells were associated with neighboring cells which were positive for either Fas ligand or TNF. However, most of the apoptotic cells were associated with TRAIL-positive cells. The nature of these TRAIL-positive cells was undetermined. Here, we report that HIV infection of primary human macrophages switches on abundant TRAIL production both at the RNA and protein levels. Furthermore, more macrophages produce TRAIL than are infected by HIV, indicating that a bystander mechanism may, at least in part, upregulate TRAIL. Exogenously supplied HIV-1 Tat protein upregulates TRAIL production by primary human macrophages to an extent indistinguishable from infection. The results suggest a model in which HIV-1-infected cells produce extracellular Tat protein, which in turn upregulates TRAIL in macrophages which then can induce apoptosis in bystander T cells. PMID- 11385303 TI - NMP 22, BTA stat test and cytology in the diagnosis of bladder cancer: a comparative study. AB - OBJECTIVE: To evaluate the usefulness of the NMP 22 and BTA stat test in the diagnosis and follow-up of bladder cancer and to compare these tests to cytology and cystoscopy, routine diagnostic methods. METHODS: 150 patients followed up for bladder cancer or symptoms suggestive of bladder cancer underwent cystoscopy after cytology, NMP 22 and BTA stat test using a recently voided urine sample. In suspect cases, TUR and histopathological analysis were performed. RESULTS: Bladder cancer was proven in 76 patients and excluded in 74. For NMP 22 we have used the cutoff value recommended by the manufacturer (10 U/ml) and that obtained by our receiver-operating characteristic curve (6 U/ml). Sensitivity was 84.21% for NMP 22 at the cutoff value of 6 U/ml and 76.32% with 10 U/ml; 72.37% for BTA stat test; 69.74% for cytology, and 100% for cystoscopy. Specificity was 86.49% for NMP 22 at a cutoff value of 6 U/ml and 90.54% at 10 U/ml; 89.19% for the BTA stat test; 93.24% for cytology and 89.19% for cystoscopy. NMP 22 sensitivity for grades 1, 2, and 3 was 68.75, 75.86 and 100%, respectively, at a cutoff value of 6 U/ml, and 50, 68.97 and 96.77%, respectively, at a cutoff level of 10 U/ml; for BTA stat the sensitivity was 56.25% in G1, 62.07% in G2 and 90.32% in G3, and for cytology the sensitivity was 43.75, 62.07 and 90.32%, respectively. The sensitivity of NMP 22 was 68.75% in stage Ta, 84.78% in T1 and 100% in T2-T4 at a cutoff level of 6 U/ml and 50, 80.43 and 92.86%, respectively, at a cutoff level of 10 U/ml; BTA stat sensitivity was 50% in Ta, 73.91% in T1 and 92.86% in T2-T4; and in cytology the results were 37.50, 73.91 and 85.71%, respectively. Using the McNemar test, there was only a significant difference between the sensitivity of NMP 22 at a cutoff level of 6 U/ml and cytology in the overall sample. CONCLUSIONS: The high sensitivity of the NMP 22 and BTA stat test in combination with the data obtained from the parameters used for the evaluation of the test demonstrate their usefulness in the diagnosis and follow-up of bladder cancer. NMP 22 at a cutoff value of 6 U/ml is significantly more sensitive than cytology and consequently a thoroughly valid diagnostic tool in the diagnosis of bladder cancer which may substitute voided urine cytology. PMID- 11385305 TI - Morbidity and discomfort of ten-core biopsy of the prostate evaluated by questionnaire. AB - Transition zone biopsies have been found to increase the detection rates of cancer of the prostate in patients with negative digital rectal examination. There are however no data available whether the higher biopsy rate is associated with greater morbidity. The present study was therefore designed to evaluate the complication rate of extended sextant biopsy. In this prospective study, 162 consecutive patients who presented for prostatic evaluation were included. After starting prophylactic antibiotic treatment 48 h prior to the procedure, transrectal ultrasound-guided core biopsies were obtained from each lobe: three each from the peripheral zone (apex, mid-zone and base) and two from the transition zone of each prostatic lobe. In all patients a questionnaire was obtained 10-12 days after the procedure. Major complications occurred in 3 patients. In 2 of the 3 cases major macroscopic hematuria was treated by an indwelling catheter for 1 or 2 days and 1 patient developed fever >38.5 degrees C for 1 day. Minor macroscopic hematuria was present in 68.5% of the patients. In 17.9% of these cases, the hematuria lasted for more than 3 days. Hematospermia was observed in 19.8% and minor rectal bleeding occurred in 4.9%. Ten-core biopsies did not lead to an increase in adverse effects or complications when compared to the results of sextant biopsies reported in the literature. PMID- 11385304 TI - Transitional zone and anterior peripheral zone of the prostate. A correlation of small-volume cancer in the biopsy cores and high psa with positive anterior margins in radical prostatectomy specimens. AB - INTRODUCTION: Prognostically significant prostatic adenocarcinomas (PAC) may pose diagnostic problems if they were localized in the anterior peripheral zone (APZ) or transitional zone (TZ). MATERIALS AND METHODS: 108 cases of PAC were reviewed along with serum PSA and TRUS biopsies. The PACs were divided into 22 TZ, 17 APZ and 69 posterior peripheral zone (PPZ) PACs according to the location of the main tumor mass in the TZ and anterior or posterior half of the peripheral zone in the radical prostatectomy (RP) specimens. RESULTS: In comparison with PPZ PAC, TZ PAC had a higher cancer volume in RP specimens (4 +/- 2.1 vs. 2.5 +/- 1.7 cm3, p < 0.01), a higher serum PSA (16.5 +/- 9.8 vs. 8.4 +/- 4.5 microg/l, p < 0.001), a biopsy with a small cancer volume (3.8 +/- 2.1 vs. 11.8 +/- 9.4 mm, p < 0.005), and a lower Gleason's score (4.8 +/- 2.1 vs. 6.5 +/- 1.7). APZ PAC was characterized by the cancer volume in RP and biopsy and PSA intermediate between those of TZ and PPZ PAC. Among 24 PACs with a total cancer core length of <3 mm, 19 cases were from the TZ and APZ groups and also had a higher cancer volume and PSA than those from the PPZ group (2.9 +/- 1.8 vs. 1.5 +/- 1.3 and 13.7 +/- 8.3 vs. 9.6 +/- 4 microg/l, respectively). Furthermore, there was a better correlation coefficient (r(2)) of tumor volume in the biopsy and RP for PPZ than for all zones PAC (r2 = 0.75 vs. 0.29). TZ and APZ carcinomas were associated with extension or satellite nodules of PAC in the PPZ that may be diagnosed with biopsies. These PACs were associated with positive anterior resection margin due to extracapsular extension of the carcinoma or intracapsular dissection in 6 and 5 cases respectively. CONCLUSIONS: TZ and APZ PACs accounted for the poor correlation between the tumor volume in the biopsy and the RP, and were associated with positive anterior resection margins. One core biopsy with a total cancer core length of <3 mm and PSA >10 microg/l are suspicious for TZ and APZ PCA in patients with undetectable tumors with DRE or TRUS. Clinically insignificant PACs tend to be associated with cancer core <3 mm and PSA <10 microg/l. PMID- 11385306 TI - The applicability of using straight ureteral stents for the treatment of ureteral stones in presumably non-compliant patients. AB - There have been many reports describing the complications of retained ureteral stents following stone treatment. We wanted to evaluate the practicality of definitive treatment of poorly compliant patients who present with ureteral stones using a straight stent connected to a urethral catheter alone and compared these to patients treated with double-J stents alone. We treated 23 patients (12 in group I and 11 patients in group II) who had a ureteral stone of 6 mm or less, with an indwelling straight stent and a double-J stent, respectively, while on oral antibiotics. We followed these patients 1 week later with an abdominal X-ray prior to removing the stent. Eleven patients in group I and 9 patients in group II passed their stones spontaneously. Three patients required surgical intervention with a ureterscope and laser lithotripsy. There were no cases of infection or pyelonephritis. Although each of the straight-stent-treated patients returned to our clinic for follow-up, only 5 of the 11 double-J stent patients returned for follow-up. The remaining 6 patients had to be contacted to remind them that they still had an internal stent. Although technologic advances now allow many urologists to definitively treat ureteral stones, some urologists may lack the proper equipment and/or assistance to treat the stone at the time of presentation, or may deal with non-compliant patients. Therefore, in these certain circumstances, treatment of small ureteral stones in non-compliant patients using a straight stent connected to a leg bag, as either definitive or initial treatment, may be of practical use and avoid the risk of retained double J ureteral stents. PMID- 11385307 TI - Intracellular electrical activity in human urinary bladder smooth muscle: the effect of high sucrose medium. AB - INTRODUCTION: The primary key to pharmacotherapy of bladder instability is in the excitation-contraction coupling of detrusor smooth muscle cells. To study this process, simultaneous recordings of mechanical and electrical activity are required. However, recording of mechanical activity induces movement, which may affect the quality of intracellular recordings. MATERIALS AND METHODS: We therefore compared the electrical activity of human detrusor smooth muscle cells in normal Krebs' solution and in a hypertonic solution, which immobilizes the tissue, enabling us to study the effect of movement on the membrane potential. Carbachol and KCl were applied to induce contractions. RESULTS: Sucrose in the medium made the tissue rigid and abolished its movement, while the electrical response was not affected. When compared with recordings in normal Krebs' solution, the average resting membrane potential was not altered. However, the membrane potential was more stable, with far less spike-shaped potentials. The spike-shaped potential amplitude was larger, while the duration was decreased. CONCLUSIONS: Impairing the ability of tissue movement resulted in changes in the electrophysiological properties of detrusor smooth muscle cells. The results suggest that stretch has an effect on L-type Ca2+ channels. PMID- 11385308 TI - Prostatic malacoplakia associated with prostatic abscess: diagnosis and treatment. AB - Prostatic malacoplakia associated with prostatic abscess is an extremely rare disease. We present a case of prostatic malacoplakia presenting as a prostatic and seminal vesicle abscess in a patient with diabetes. The diagnosis and management are discussed, and the literature is reviewed. PMID- 11385309 TI - Primary squamous cell carcinoma of the prostate: a rare clinicopathological entity. Report of 2 cases and review of literature. AB - Primary squamous cell carcinoma of the prostate is an uncommon clinicopathological entity. It differs from more common adenocarcinomas in its cell of origin, biological behavior, therapeutic response to the usual hormonal manipulation and prognosis. The review shows that squamous cell carcinoma is biologically more aggressive than adenocarcinoma. Despite the agreement on its uniqueness, a controversy exists on the exact histopathogenesis, diagnostic criteria and modality of treatment. We report on 2 patients with primary squamous cell carcinoma of the prostate. One patient presented with lower urinary tract symptoms with a hard nodular prostate on digital rectal examination, and the other with acute urinary retention on normal digital rectal examination. The evaluation revealed metastasis in the pelvic and right femur in both cases. Palliative transurethral resection of the prostate with chemotherapy (Adriamycin based) was given in both the cases. However, both the patients died at 4 and 5 months of follow-up, respectively. PMID- 11385310 TI - MALT-type primary lymphoma of the urinary bladder: clinicopathological study of 2 cases and review of the literature. AB - Primary lymphoma of the bladder is a rare non-epithelial bladder tumour. It is usually non-Hodgkin's lymphoma with mucosa-associated lymphoid tissue (MALT) lymphoma being its predominant subtype. Most of the literature has been limited to individual sporadic case reports. We present 2 cases of the MALT-type primary lymphoma of the urinary bladder which were treated in our unit. Both patients were females with a history of recurrent urinary tract infections. They were treated successfully with chemotherapy alone. We also present a review of the literature emphasising the clinical presentation, the morphological and immunophenotypic features and the various treatment modalities of this rarely seen bladder pathology. PMID- 11385311 TI - An unusual cause of irritable urinary bladder symptoms. AB - A metastasis from a renal cell carcinoma to a seminal vesicle is extremely rare. This pattern of metastatic disease has been reported once previously in the Japanese literature. We describe a patient with irritability symptoms of the urine bladder and involuntary loss of stool caused by obstruction of the bladder by a large metastasis from a renal cell carcinoma to the seminal vesicle. The mode of diagnosis and treatment of this patient are described. PMID- 11385312 TI - Transvesical placement of one limb of an aortobifemoral bypass graft as a complication of aortic bypass surgery. AB - We report about a rare complication after aortobifemoral bypass surgery. In the present case the left limb of an aortobifemoral bypass graft was placed straight through the cavity of the urinary bladder. The diagnosis was found and ascertained by colour duplex ultrasound examination, cystoskopy and angiography. In consequence, we successfully performed open bladder surgery to replace the misplaced graft. PMID- 11385313 TI - Handicap of walking by a huge paratesticular liposarcoma. AB - We describe the case of a 71-year-old male with a huge left-sided paratesticular tumour, whose walking was increasingly handicapped by this vast mass. Two palliative excisions of tumour tissue were performed. Histology revealed a poorly differentiated paratesticular liposarcoma. The patient achieved satisfying mobility for several months before he died of cachexia. PMID- 11385314 TI - IgH, TCR-gamma, and TCR-beta gene rearrangement in 80 B- and T-cell non-Hodgkin's lymphomas: study of the association between proliferation and the so-called "aberrant" patterns. AB - The current study analyzes the rearrangement pattern of immunoglobulin H (IgH), T cell receptor (TCR)-gamma, and TCR-beta genes in a group of 80 non-Hodgkin's lymphomas (NHL) of different histologic subtypes (43 B-cell and 37 T-cell types). The sensitivity and specificity provided by polymerase chain reaction amplification of these loci are evaluated. The association between the proliferation index and the presence of the so-called "aberrant" or "dual" rearrangements is also considered. Ninety-one percent of B-cell NHL showed IgH gene monoclonality, and 21% also exhibited a monoclonal pattern in one of the TCR genes. Among T-cell NHL, the sensitivity of the study was 65% for the TCR-gamma gene and 46% for the TCR-beta gene. The total sensitivity was 76%, amplifying both loci. IgH gene aberrant rearrangements were observed in 16% of T-cell neoplasms. A substantial percentage of dual rearrangements were detected in precursor and mature B- and T-cell NHL. B-cell NHL showed a tendency toward higher values of proliferation when aberrant rearrangements were present; however, this trend was not significant. Furthermore, in the case of T-cell NHL there was a significant negative association between these two variables. PMID- 11385315 TI - The value of clonality in the diagnosis and follow-up of patients with cutaneous T-cell infiltrates. AB - The diagnosis of early stages of cutaneous T-cell lymphoma (CTCL) is often difficult, especially for lesions that are at the borderline between reactive and neoplastic skin T-cell infiltrates. T-cell monoclonality in these lesions is considered by some to be an important prognostic factor of neoplastic evolution, whereas others claim that clonality can also be found in benign skin infiltrates and is therefore of limited diagnostic value. To address this controversy, the authors analyzed retrospectively eight patients with lymphocytic skin lesions who progressed to CTCL, and three patients with recurrent T-cell lymphocytic infiltrates who had not developed CTCL. From a total of 65 biopsies of eight progressing patients, 32 were diagnosed as histologically malignant and 33 were diagnosed as benign or borderline. The authors found clonality by either polymerase chain reaction or Southern blot analysis in 88% of malignant and in 79% of nonmalignant lesions. None of the 37 biopsies of non-progressing patients was clonal. These results indicate strongly that the presence of monoclonality in T-cell skin infiltrates is related closely to the risk of developing CTCL. The value of clonality as a marker of malignancy is supported by the absence of T cell clonal populations in all infiltrates from patients who had not progressed to lymphoma. PMID- 11385317 TI - Kaposi sarcoma-associated herpesvirus/human herpesvirus 8 and multiple myeloma in South Africa. AB - Kaposi sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8) has been implicated in the etiopathogenesis of multiple myeloma. Although the association is biologically plausible and attractive, conflicting data have been reported, including evidence against the involvement of KSHV in the pathogenesis of the disease. The purpose of this study was to determine the relationship between KSHV and myeloma in blacks in South Africa, in whom the disease is not uncommon and the seroprevalence of KSHV is higher than in the areas in which this association has been documented. Using a nested polymerase chain reaction (PCR) assay, the authors initially tested for the presence of KSHV DNA sequences (KS330(233)) in bone marrow aspirates, bone marrow biopsy material, and cultured bone marrow adherent cell samples of patients with myeloma. KSHV DNA sequences were detected in 4 of 10 (40%) of the adherent cell cultures and 1 of 20 (5%) of the bone marrow aspirate samples. None of the bone marrow biopsy samples (0/9) or control bone marrow aspirate samples (0/19) was positive. To confirm the positive results in the bone marrow cultures noted above and to exclude contamination, the procedure was repeated in a further 7 patients with myeloma and 11 controls with lymphoproliferative disorders using the same nested PCR assay. In addition, the authors used a different set of primers that recognize sequences internal to the 233-bp fragment to yield a final product of 186 bp. The authors were unable to detect any KSHV DNA sequences in the patients with myeloma (0/7) or the control patients with other lymphoproliferative disorders (0/11). Taken together, the finding of a positive result in 4 of 17 patients (23.5%), which is similar to the background seroprevalence rate, does not support a clear association between myeloma and KSHV in blacks in South Africa. PMID- 11385316 TI - A 1-kb Bcl-2-PCR fragment detection in a patient with follicular lymphoma and development of a new diagnostic PCR method. AB - The t(14;18) translocation is a useful marker to characterize follicular lymphoma and to monitor residual disease. The polymerase chain reaction (PCR) is a powerful technique to detect this translocation. Located on chromosome 18, within the Bcl-2 gene, breakpoints occur mainly in the 3; untranslated region, in the third exon of Bcl-2 (MBR region). In this study, the authors amplified MBR breakpoints by PCR and found an unexpectedly large fragment of 1 Kb that corresponds to a recently described new breakpoint in the Bcl-2 gene. With a new primer set, a further previously considered t(14;18)-unrelated tumor was in fact positive for this new breakpoint. PMID- 11385319 TI - A multiethnic study of Delta32ccr5 and ccr2b-V64I allele distribution in four Los Angeles populations. AB - Mutant alleles of the chemokine receptors CCR5 and CCR2 affect the susceptibility to HIV infection as well as the rate of disease progression. In this article the authors report the results of a survey for presence of the common Delta32ccr5 and ccr2b-V64I mutant alleles in 472 individuals of a multiethnic cohort. Hispanic Americans had the highest observed frequency of the Delta32ccr5 allele (3.57%), whereas African Americans had a lower frequency (1.55%). The mutant allele was absent in Asian Americans and Native Americans. Thus, the Delta32ccr5 allele segregates in populations with a significant white admixture and is rare in genetically distant non-European groups. Native Americans had the highest occurrence of the ccr2b-V64I allele (31.13%), whereas African Americans, Asian Americans, and Hispanic Americans had much lower frequencies (14.36%, 11.94%, and 14.37% respectively). This mutation is probably an ancient one, occurring before the migration of the ancestors of Native Americans across the Bering Straits to the Americas. The twofold greater frequency of ccr2b-V64I in modern Native Americans probably reflects a founder effect. The observed population differences in Delta32ccr5 and ccr2b-V64I frequencies, considered together with their documented effects on sensitivity to HIV infection and rate of disease progression, have implications for HIV transmission patterns in the United States, as well as for AIDS prediction, monitoring, and treatment. PMID- 11385318 TI - Rearrangement in the coding region of the MYCN gene in a subset of amplicons in a case of neuroblastoma with MYCN amplification. AB - The MYCN gene is often amplified but rarely rearranged in neuroblastoma. We report, for the first time, a rearrangement within the MYCN coding region in a metastatic neuroblastoma in a 3-year-old boy with MYCN amplification in his primary tumor. The rearrangement occurred 46 nucleotides downstream from the ATG codon in exon 2 of MYCN. The amplification level of the rearranged copies of the MYCN gene was lower than that of the unrearranged copies of MYCN. These results indicate that the rearrangement occurred after initial MYCN gene amplification. Monochromosomal somatic cell hybrid mapping of the novel region fused to exon 2 of MYCN localized it to chromosome 2, suggesting that this rearrangement resulted from an interstitial deletion, presumably within the MYCN amplicon itself. PMID- 11385320 TI - Coexistence of the methylenetetrahydrofolate reductase single-nucleotide polymorphism (C677T) in patients with the factor V Leiden or prothrombin G20210A polymorphisms. AB - Venous thromboembolism is a complex disease resulting from the interaction of a multitude of both genetic and environmental factors that can affect the cascade of biochemical reactions involved. Single-nucleotide polymorphisms in the genes that code for coagulation factors V (factor V Leiden) and II (prothrombin G20210A), as well as the methylenetetrahydrofolate reductase (MTHFR C677T) gene, have been implicated in the majority of cases of hereditary thrombophilia. In this study the authors evaluated the coexistence of the MTHFR polymorphism in 96 patients with a clinical suspicion for thrombosis who also have either the factor V Leiden polymorphism or the prothrombin G20210A polymorphism. Results indicate that the frequency of the MTHFR polymorphism was similar between the study and control groups with respect to heterozygosity (36.5% vs. 55.3%) and homozygosity (20.8% vs. 14.9%). These data suggest that the MTHFR polymorphism is not associated preferentially with patients who have had or who are at risk of developing a thrombotic event. In this study, patients with the factor V Leiden polymorphism or the prothrombin G20210A polymorphism were considered to be at risk. PMID- 11385321 TI - beta-Catenin expression pattern, beta-catenin gene mutations, and microsatellite instability in endometrioid ovarian carcinomas and synchronous endometrial carcinomas. AB - beta-Catenin gene mutations and microsatellite instability (MI) have been reported in endometrioid ovarian carcinomas. In colon but not endometrial cancer, beta-catenin gene mutations are associated with a replication error phenotype and MI. In this study the authors investigate whether beta-catenin mutations and MI are two independent oncogenic pathways in endometrioid ovarian carcinomas. They also evaluate the usefulness of these molecular markers in determining the primary origin of simultaneous tumors in the ovary and endometrium. This study was performed on 26 patients diagnosed with primary endometrioid ovarian carcinoma, five of whom also had pathologically diagnosed primary synchronous endometrioid endometrial carcinoma. Immunohistochemical and molecular analyses indicated that there were 25 primary ovarian tumors with four primary synchronous endometrial cancers and one ovarian metastasis of a primary endometrial carcinoma. All studies were performed on formalin-fixed, paraffin-embedded tissue samples. The beta-catenin expression pattern (nuclear vs. membranous) was analyzed immunohistochemically. Mutations in exon 3 of the beta-catenin gene were studied by polymerase chain reaction, single-strand conformational polymorphism, and direct sequencing. MI status was established by studying BAT-26 and BAT-25 mononucleotide repeats. In the group with 21 single ovarian tumors, 18 (85%) had beta-catenin nuclear expression, eight (38%) had beta-catenin gene mutations (always associated with beta-catenin nuclear expression), and four (19%) had MI. Only one case (5%) had both beta-catenin gene mutations and MI. The mutations affected one of the serine/threonine residues targeted for phosphorylation by glycogen synthase kinase-3beta or adjacent residues. At codon 32, a GAC-to-TAC (D32Y) change was found; at codon 33, two TCT-to-TGT (S33C) changes were found; at codon 37, three TCT-to-TTT (S37F) changes and one TCT-to-TGT (S37C) change were found; and, lastly, one ACC-to-GCC change at codon 41 (T41A) was detected. Four of the 25 endometrioid ovarian carcinomas (16%) had an associated synchronous endometrial carcinoma. There was a higher percentage of beta-catenin mutations (n = 3, 75%) in synchronous ovarian carcinomas than in single ones, although with a similar percentage of MI (n = 1, 25%). beta-catenin mutations were S37C in two cases and D32G in one. One of the four endometrial carcinomas showed an S33C beta-catenin mutation, and two carcinomas had MI. None of the four tumors had both beta-catenin gene mutation and MI. beta-catenin gene mutations were always associated with a nuclear beta-catenin expression pattern, whereas MI was associated with a membranous pattern. In one patient both the ovarian and the endometrial carcinomas had beta-catenin gene mutations, in another patient both tumors showed MI, whereas in the remaining two patients the ovarian carcinomas showed beta-catenin gene mutations and the endometrial carcinomas showed MI. To summarize, the results of this study suggest that beta-catenin mutations and MI could represent two independent pathways in endometrioid ovarian carcinomas because they occur simultaneously very infrequently (in 5% of these cases). beta catenin mutations are always associated with a nuclear beta-catenin expression pattern, whereas cases with a replication error -plus phenotype showed no abnormal beta-catenin subcellular localization. The study of the beta-catenin expression pattern, beta-catenin mutations, and MI, together with conventional clinicopathologic findings, could aid in distinguishing between the metastatic or independent origin of simultaneous endometrioid ovarian and endometrial carcinomas. Tumors with identical immunohistochemical and molecular features should therefore be considered to have a common origin. PMID- 11385322 TI - Human telomerase reverse transcriptase expression in Diff-Quik-stained FNA samples from thyroid nodules. AB - Fine-needle aspiration (FNA) is a highly sensitive method in the differential diagnosis of thyroid nodules. However, 10% of thyroid FNAs are indeterminate for cancer, and thus additional markers may be useful diagnostically. The authors have demonstrated previously that human telomerase reverse transcriptase (hTERT) gene expression is useful in the distinction of benign lesions from malignant lesions. They therefore wondered whether the detection of hTERT gene expression was feasible using archival slides. To establish an experimental system, ribonucleic acid was extracted from human anaplastic thyroid carcinoma cell line (ARO) in cytologic specimens, and reverse transcription-polymerase chain reaction (RT-PCR) for hTERT expression was performed. RT-PCR analysis for hTERT gene detection was then performed using 58 Diff-Quik-stained archival FNA samples collected retrospectively. RT-PCR for human thyroglobulin (hTg) or beta-actin gene expression served as a positive control. Successful PCR results were obtained from 48 of the 58 cases. All 10 slides in which no RT-PCR products were noted were older than 3 years. hTERT gene expression was demonstrated in FNAs from two of seven cases (29%) of hyperplastic nodule, one of one case (100%) of Hashimoto's thyroiditis, three of eight cases (38%) of follicular adenoma, three of eight cases (38%) of Hurthle cell adenoma, three of four cases (75%) of follicular carcinoma, two of two cases (100%) of Hurthle cell carcinoma, and 11 of 18 cases (61%) of papillary carcinoma. All but one of the available 33 corresponding frozen samples exhibited the same RT-PCR results. This study demonstrates that Diff-Quik-stained thyroid FNA specimens less than 3 years old can be used for the detection of hTERT gene expression by RT-PCR. This test, along with careful cytopathologic examination, may improve our ability to differentiate benign lesions from malignant lesions in indeterminate FNA samples from thyroid nodules. PMID- 11385323 TI - A rapid PCR method for the detection of slime-producing strains of Staphylococcus epidermidis and S. aureus in periprosthesis infections. AB - In periprosthesis tissues, Staphylococcus epidermidis produces extracellular polysaccharide slime. Recently it has been shown that S. aureus also produces slime and that both S. epidermidis and S. aureus contain the ica operon responsible for slime production. In the operon, icaA encodes for N acetylglutaminyltransferase, the enzyme for polysaccharide synthesis. However, co expression of icaA and icaD is required for full slime synthesis. The slime producing strains of both S. epidermidis and S. aureus are more virulent and are responsible for severe postsurgical or periprosthesis infections. The authors describe a simple, rapid, and reliable polymerase chain reaction method to detect icaA and icaD. The method was applied to the detection of ica genes on two reference strains, 15 strains each of S. epidermidis and S. aureus from periprosthesis infections and 10 strains from the skin and mucosa of healthy volunteers. icaA and icaD were detectable only in slime-producing strains (tested for slime production on Congo Red agar), and never in nonslime-producing ones. This method is a straightforward way of detecting the slime-producing ability by S. epidermidis and S. aureus. In clinical specimens this polymerase chain reaction method enables rapid diagnosis of virulent slime-producing strains with respect to the traditional culture method on Congo Red agar, which requires much more time. Rapid identification of the virulent properties of the bacterial strain responsible for a staphylococcal infection is crucial for deciding treatment. PMID- 11385324 TI - Frequent use of the hospital emergency department is indicative of high use of other health care services. AB - STUDY OBJECTIVE: We sought to determine the proportion of emergency department patients who frequently use the ED and to compare their frequency of use of other health care services at non-ED sites. METHODS: A computerized patient database covering all ambulatory visits and hospital admissions at all care facilities in the county of Stockholm, Sweden, was used. Frequent ED patients were defined as those making 4 or more visits in a 12-month period. RESULTS: Frequent users comprised 4% of total ED patients, accounting for 18% of the ED visits. The ED was the only source of ambulatory care for 13% of frequent versus 27% of rare ED users (1 ED visit). Primary care visits were made by 72% of frequent ED users versus 57% by rare ED visitors. The corresponding figures for hospital admission were 80% and 36%, respectively. Frequent ED visitors were also more likely to use other care facilities repeatedly: their odds ratio (adjusted for age and sex) was 3.43 (95% confidence interval [CI] 3.10 to 3.78) for 5 or more primary care visits and 29.98 (95% CI 26.33 to 34.15) for 5 or more hospital admissions. In addition, heavy users had an elevated mortality (standardized mortality ratio 1.55; 95% CI 1.26 to 1.90). CONCLUSION: High ED use patients are also high users of other health care services, presumably because they are sicker than average. A further indication of serious ill health is their higher than expected mortality. This knowledge might be helpful for care providers in their endeavors to find appropriate ways of meeting the needs of this vulnerable patient category. PMID- 11385325 TI - Inappropriate use of an accident and emergency department: magnitude, associated factors, and reasons--an approach with explicit criteria. AB - BACKGROUND: We evaluate the appropriateness of medical visits to the accident and emergency department (A&ED) of a university hospital using an instrument based on explicit and objective criteria, analyze the association between inappropriate visits and certain factors, and identify reasons for inappropriate use. METHODS: This concurrent review of a random sample of 2,980 adult medical patients' visits to the A&ED of the hospital of Elche uses the Hospital Urgencies Appropriateness Protocol, an instrument based on explicit criteria. We analyze the association between inappropriate use and specific factors, and provide a descriptive analysis of reasons for inappropriate use assigned by A&ED staff. RESULTS: Of the total number, 882 (29.6%) of the visits were evaluated as inappropriate. Inappropriate use was associated with younger patients, use of own means of transportation, referral by the hospital, certain months of the year, and certain diagnostic groups of lesser severity. The most frequent reasons for inappropriate use were the patients' greater trust in the hospital than primary care (451 [51.1%]), inappropriate use of services by patients (160 [18.1%]), and inappropriate referrals by primary care physicians (142 [16.1%]). CONCLUSION: Inappropriate use represents an important percentage of use of the A&ED. Many reasons contribute to it, although foremost among them is patient preference (and the convenience and accessibility) of these services compared with primary care. PMID- 11385326 TI - Appropriateness of emergency department visits in a Portuguese university hospital. AB - STUDY OBJECTIVE: There are no studies in Portugal supporting a common claim that most emergency department visits are inappropriate. The aim of this study was to determine the prevalence of and to evaluate factors associated with an appropriate ED visit in a major public hospital. METHODS: A cross-sectional prospective study was performed at a public university hospital ED. Data for demographic variables, duration of complaint, transfer from other medical sources, and previous medical care for the same complaints were collected by interviewing all patients who arrived at the ED within a consecutive period of at least 24 hours. Data for diagnostic tests, treatment performed, and final patient destination were collected by triage records review. An appropriate ED visit was defined by explicit criteria: interhospital transfer, patient death at the ED, hospitalization, and diagnostic tests or treatments performed. RESULTS: The study included 5,818 adult patients. The prevalence of an appropriate ED visit, by use of our criteria, was 68.7%. Sex was an effect modifier. According to this study, determinants of an appropriate visit for men and women were age 60 years or older and complaints of 24 hours or less and in women but not in men, retired from work and with arrival between midnight and 8 AM. CONCLUSIONS: In a university hospital in Oporto, the majority of ED visits were appropriate according to explicit criteria. Some variables may be associated with appropriateness of ED visits. A duration of the complaint 24 hours or less along with an arrival between midnight and 8 AM in women and age 60 years or older in men were the most important determinants. PMID- 11385327 TI - Executive summary: developing objectives, content, and competencies for the training of emergency medical technicians, emergency physicians, and emergency nurses to care for casualties resulting from nuclear, biological, or chemical incidents. AB - STUDY OBJECTIVE: The task force assessed the needs, demands, feasibility, and content of training for US civilian emergency medical responders (paramedics, nurses, and physicians) for nuclear/biological/chemical (NBC) terrorism. METHODS: A task force representing key professional organizations, stakeholders, and disciplines involved in emergency medical response conducted an iterated instructional-design analysis on the feasibility and content of such training with input from educational professionals. We then analyzed 6 previously developed training courses for their congruence with our recommendations. RESULTS: The task force produced descriptions of learning groups, content and learning objectives, and barriers and challenges to NBC education. Access to training and sustainment of learning (retention of knowledge) represent the significant barriers. The courses analyzed by the task force did not meet all objectives and challenges addressed. CONCLUSION: The task force recommends training programs and materials need to be developed to overcome the identified barriers and challenges to learning for these audiences. Furthermore, the task force recommends incorporating NBC training into standard training programs for emergency medical professionals. PMID- 11385328 TI - CPR-only survivors of out-of-hospital cardiac arrest: implications for out-of hospital care and cardiac arrest research methodology. AB - STUDY OBJECTIVE: There is little evidence that cardiopulmonary resuscitation (CPR) alone may lead to the resuscitation of cardiac arrest victims with other than respiratory causes (eg, pediatric arrest, drowning, drug overdose). The objective of this study was to identify out-of-hospital cardiac arrest survivors resuscitated without defibrillation or advanced cardiac life support. METHODS: This observational cohort included all adult survivors of out-of-hospital cardiac arrest of a cardiac cause from phases I and II of the Ontario Prehospital Advanced Life Support Study. During the study period, the system provided a basic life support/defibrillation level of care but no advanced life support. CPR-only patients were patients determined to be without vital signs by EMS personnel who regained a palpable pulse in the field with precordial thump or CPR only and then were admitted alive to the hospital. Six members of a 7-member expert review panel had to rate the patient as either probably or definitely having an out-of hospital cardiac arrest, and a rhythm strip consistent with a cardiac arrest rhythm had to be present to be considered a patient. Criteria considered were witness status, citizen or first responder CPR, CPR duration, arrest rhythm and rate, and performance of precordial thump. RESULTS: From January 1, 1991, to June 30, 1997, 9,667 patients with out-of-hospital cardiac arrest were treated. The overall survival rate to hospital discharge was 4.6%. There were 97 apparent CPR only patients admitted to the hospital. Application of the inclusion criteria yielded 24 CPR-only patients who had true out-of-hospital cardiac arrest and 73 patients judged not to have cardiac arrest. Of the 24 true CPR-only patients admitted to the hospital, 15 patients were discharged alive, 10 patients were witnessed by bystanders, and 7 patients were witnessed by EMS personnel. The initial arrest rhythm was pulseless electrical activity in 9 patients, asystole in 12 patients, and ventricular tachycardia in 3 patients. One patient with ventricular tachycardia converted to sinus tachycardia with a single precordial thump. CONCLUSION: CPR-only survivors of true out-of-hospital cardiac arrest do exist; some victims of out-of-hospital cardiac arrest of primary cardiac cause can survive after provision of out-of-hospital basic life support care only. However, many patients found to be pulseless by means of out-of-hospital evaluation likely did not have a true cardiac arrest. This has implications for the survival rates of most, if not all, previous cardiac arrest reports. Survival rates from cardiac arrest may actually be lower if one excludes survivors who never had a true arrest. The absence of vital signs by out-of-hospital assessment alone is not adequate to include patients in research reports or quality evaluations for cardiac arrest. PMID- 11385329 TI - Can an out-of-hospital cervical spine clearance protocol identify all patients with injuries? An argument for selective immobilization. AB - STUDY OBJECTIVE: We sought to determine the sensitivity of the Fresno/Kings/Madera emergency medical services (EMS) selective spine immobilization protocol in identifying patients with potential cervical injuries. We also sought to determine whether the protocol was safe in the out-of-hospital setting. METHODS: We conducted a retrospective chart review of all patients discharged from 5 trauma-receiving hospitals in Fresno County with the diagnosis of cervical spine injury between July 1, 1990, and June 30, 1996. All of these patients transported to the hospital by EMS personnel were selected for the study group. Medical records of those patients not immobilized were further investigated to identify protocol violations or deficiencies. RESULTS: There were 861 patients with significant cervical injuries during this time span. EMS personnel brought 504 patients to the hospital, of which 495 arrived in cervical spine immobilization. Of the remaining 9 patients, 2 refused immobilization, and 2 could not be immobilized; 3 injuries were missed by the protocol criteria, and 2 injuries were missed because of protocol violations. Of these last 5 patients, 1 patient had an adverse outcome, 2 injuries were considered unstable, 4 patients were older than 67 years, and one patient was 9 months old. CONCLUSION: The Fresno/Kings/Madera EMS selective spine immobilization protocol is 99% (95% CI, 97.7% to 99.7%) sensitive in identifying patients with cervical injuries for immobilization. Those patients not identified were at extremes of age. These results suggest that selective immobilization may be safely applied in the out-of hospital setting but should be used with caution at extremes of age. PMID- 11385330 TI - Probabilistic linkage of computerized ambulance and inpatient hospital discharge records: a potential tool for evaluation of emergency medical services. AB - STUDY OBJECTIVES: Emergency medical services (EMS) is an important part of the health care system. The effect of EMS on morbidity, mortality, and costs of illness is difficult to evaluate because hospital information is not available in out-of-hospital databases. We used probabilistic linkage to create such a database from ambulance and inpatient data and demonstrate the potential for linkage to facilitate evaluation of EMS responses resulting in hospital admission. METHODS: Statewide ambulance and inpatient hospital discharge records were available for 1994 through 1996. Ambulance records indicating admission to the emergency department or hospital (165,649 records) were linked to inpatient hospital records indicating emergency admission (146,292 records) by using probabilistic linkage. Out-of-hospital data (dispatch code, treatments rendered, and ages), linkage rates, and inpatient data (discharge status, charges, length of stay, and payer category) were analyzed. RESULTS: We linked 24,299 (14.7%) ambulance events to inpatient hospital discharges. If we had used exact linkage methods, we would have only linked 14,621 record pairs, a loss of nearly 40%. Linkage rates were relatively constant between years (approximately 15%) but differed by ambulance dispatch codes. Out-of-hospital dispatch codes with high linkage rates included breathing problems (22.6%), chest pain (21.5%), diabetic problems (16.9%), drowning incidents (14.9%), falls (19.2%), strokes (32.8%), and unconsciousness or fainting episodes (16.1%). Linkage to the hospital record provided access to hospital outcome data. Inpatient mortality was 6.8%. Survivors were discharged home (60.7%), transferred to other acute-care facilities (3.6%) or intermediate-care facilities (23.3%), or discharged with home health care provision (4.9%). The median length of stay was 3 days, and median charges were $6,620; total inpatient charges were $286,737,067. CONCLUSION: Probabilistic linkage enables ambulance and hospital discharge records to be linked together and potentially increases our ability to critically evaluate EMS by providing access to hospital-based outcomes. Such evaluation will be further improved by linking to ED, other outpatient, and other public health data sources. PMID- 11385331 TI - Frequent emergency department use in Sweden: implications for emergency medicine in the United States. PMID- 11385332 TI - Appropriate standards for "appropriateness" research. PMID- 11385333 TI - Out-of-hospital cervical spine immobilization: making policy in the absence of definitive information. PMID- 11385334 TI - Hydroxocobalamin: improved public health readiness for cyanide disasters. AB - The United States is under the constant threat of a mass casualty cyanide disaster from industrial accidents, hazardous material transportation incidents, and deliberate terrorist attacks. The current readiness for cyanide disaster by the emergency medical system in the United States is abysmal. We, as a nation, are simply not prepared for a significant cyanide-related event. The standard of care for cyanide intoxication is the cyanide antidote kit, which is based on the use of nitrites to induce methemoglobinemia. This kit is both expensive and ill suited for out-of-hospital use. It also has its own inherent toxicity that prevents rapid administration. Furthermore, our hospitals frequently fail to stock this life-saving antidote or decline to stock more than one. Hydroxocobalamin is well recognized as an efficacious, safe, and easily administered cyanide antidote. Because of its extremely low adverse effect profile, it is ideal for out-of-hospital use in suspected cyanide intoxication. To effectively prepare for a cyanide disaster, the United States must investigate, adopt, manufacture, and stockpile hydroxocobalamin to prevent needless morbidity and mortality. PMID- 11385335 TI - Terrorism and the ethics of emergency medical care. AB - The threat of domestic and international terrorism involving weapons of mass destruction-terrorism (WMD-T) has become an increasing public health concern for US citizens. WMD-T events may have a major effect on many societal sectors but particularly on the health care delivery system. Anticipated medical problems might include the need for large quantities of medical equipment and supplies, as well as capable and unaffected health care providers. In the setting of WMD-T, triage may bear little resemblance to the standard approach to civilian triage. To address these issues to the maximum benefit of our patients, we must first develop collective forethought and a broad-based consensus that these decisions must reach beyond the hospital emergency department. Critical decisions like these should not be made on an individual case-by-case basis. Physicians should never be placed in a position of individually deciding to deny treatment to patients without the guidance of a policy or protocol. Emergency physicians, however, may easily find themselves in a situation in which the demand for resources clearly exceeds supply. It is for this reason that emergency care providers, personnel, hospital administrators, religious leaders, and medical ethics committees need to engage in bioethical decision making before an acute bioterrorist event. PMID- 11385336 TI - Hospital trauma care in multiple-casualty incidents: a critical view. AB - During a multiple-casualty incident, a large casualty caseload adversely affects the quality of trauma care given to individual patients. From a trauma care perspective, the goal of the hospital emergency plan is to provide severely injured patients with a level of care that approximates the care given to similar patients under normal conditions. Therefore, the realistic admitting capacity of the hospital is determined primarily by the number of trauma teams that the hospital can recruit. Effective triage of these casualties is often not straightforward, with high overtriage rates. Simplified triage algorithms may be a practical alternative to more elaborate schemes. The concept of minimal acceptable care is the key to a staged management approach during a mass-casualty incident. Discrete-event computer simulation and war game tabletop exercises for key personnel are 2 new modalities that are supplementing the traditional mock disaster drill as effective planning and training tools. PMID- 11385337 TI - Exposure to liquid sulfur mustard. AB - Chemical weapons continue to pose a serious threat to humanity. With the use of chemical weapons by terrorists in Tokyo, and the projected disarming of the chemical weapon stockpile in this country, the possibility that emergency physicians will encounter patients contaminated by chemical munitions, such as sulfur mustard, exists. Mustard is a vesicating agent with a long latency between exposure and symptoms. Exposure can cause burns, conjunctivitis, pneumonia, and death. We describe 3 workers exposed to mustard at a chemical weapon storage facility. This article reports the first case of an exposure to mustard at a storage facility, as well as the first documented incident occurring in the United States. All physicians who manage patients in an acute care setting should be aware of the presentation and emergency treatments involving patients contaminated with mustard. PMID- 11385338 TI - Emergency Medical Services Outcomes Project (EMSOP) II: developing the foundation and conceptual models for out-of-hospital outcomes research. AB - Development of methodologically acceptable outcomes models for emergency medical services (EMS) is long overdue. In this article, the Emergency Medical Services Outcomes Project proposes a conceptual framework that will provide a foundation for future EMS outcomes research. The "Episode of Care Model" and the "Out-of Hospital Unit of Service Model" are presented. The Episode of Care Model is useful in conditions in which interventions and outcomes, especially survival and major physiologic dysfunction, are linked in a time-dependent manner. Conditions such as severe trauma, anaphylaxis, airway obstruction, respiratory arrest, and nontraumatic cardiac arrest are amenable to this methodology. The Out-of-Hospital Unit of Service Model is essentially a subunit of the Episode of Care Model. It is valuable for evaluating conditions that have minimal-to-moderate therapeutic time dependency. This model should be used when studying outcomes limited to the out-of-hospital interval. An example of this is pain management for injuries sustained in motor vehicle crashes. These models can be applied to a wide spectrum of conditions and interventions. With the scrutiny of health care expenditures ever increasing, the identification of clinical interventions that objectively improve patient outcome takes on growing importance. Therefore, the development, dissemination, and use of meaningful methodologies for EMS outcomes research is key to the future of EMS system development and maintenance. PMID- 11385339 TI - Explosions and blast injuries. AB - Powerful explosions have the potential to inflict many different types of injuries on victims, some of which may be initially occult. Flying debris and high winds commonly cause conventional blunt and penetrating trauma. Injuries caused by blast pressures alone result from complex interactions on living tissues. Interfaces between tissues of different densities or those between tissues and trapped air result in unique patterns of organ damage. These challenge out-of-hospital personnel, emergency physicians, and trauma surgeons to specifically seek evidence of these internal injuries in individuals with multiple trauma, adjust management considerations to avoid exacerbation of life threatening problems caused by the blast wave itself, and ensure appropriate disposition of these patients in possible mass-casualty situations. Knowledge of the potential mechanisms of injury, early signs and symptoms, and natural courses of these problems will greatly aid the management of blast-injured patients. PMID- 11385340 TI - Feedback: computed tomography for subarachnoid hemorrhage. Which review should we believe regarding the diagnostic power of computed tomography for ruling out subarachnoid hemorrhage? PMID- 11385341 TI - Competency and confidence: procedures in the emergency department. PMID- 11385342 TI - Principles of appropriate antibiotic use for treatment of acute respiratory tract infections in adults: background, specific aims, and methods. AB - The need to decrease excess antibiotic use in ambulatory practice has been fueled by the epidemic increase in antibiotic-resistant Streptococcus pneumoniae. The majority of antibiotics prescribed to adults in ambulatory practice in the United States are for acute sinusitis, acute pharyngitis, acute bronchitis, and nonspecific upper respiratory tract infections (including the common cold). For each of these conditions--especially colds, nonspecific upper respiratory tract infections, and acute bronchitis (for which routine antibiotic treatment is not recommended)--a large proportion of the antibiotics prescribed are unlikely to provide clinical benefit to patients. Because decreasing community use of antibiotics is an important strategy for combating the increase in community acquired antibiotic-resistant infections, the Centers for Disease Control and Prevention convened a panel of physicians representing the disciplines of internal medicine, family medicine, emergency medicine, and infectious diseases to develop a series of "Principles of Appropriate Antibiotic Use for Treatment of Acute Respiratory Tract Infections in Adults." These principles provide evidence based recommendations for evaluation and treatment of adults with acute respiratory illnesses.This paper describes the background and specific aims of and methods used to develop these principles. The goal of the principles is to provide clinicians with practical strategies for limiting antibiotic use to the patients who are most likely to benefit from it. These principles should be used in conjunction with effective patient educational campaigns and enhancements to the health care delivery system that facilitate nonantibiotic treatment of the conditions in question. PMID- 11385344 TI - Principles of appropriate antibiotic use for acute rhinosinusitis in adults: background. AB - The following principles of appropriate antibiotic use for adults with acute rhinosinusitis apply to the diagnosis and treatment of acute maxillary and ethmoid rhinosinusitis in adults who are not immunocompromised. Most cases of acute rhinosinusitis diagnosed in ambulatory care are caused by uncomplicated viral upper respiratory tract infections. Bacterial and viral rhinosinusitis are difficult to differentiate on clinical grounds. The clinical diagnosis of acute bacterial rhinosinusitis should be reserved for patients with rhinosinusitis symptoms lasting 7 days or more who have maxillary pain or tenderness in the face or teeth (especially when unilateral) and purulent nasal secretions. Patients with rhinosinusitis symptoms that last less than 7 days are unlikely to have bacterial infection, although rarely some patients with acute bacterial rhinosinusitis present with dramatic symptoms of severe unilateral maxillary pain, swelling, and fever. Sinus radiography is not recommended for diagnosis in routine cases. Acute rhinosinusitis resolves without antibiotic treatment in most cases. Symptomatic treatment and reassurance is the preferred initial management strategy for patients with mild symptoms. Antibiotic therapy should be reserved for patients with moderately severe symptoms who meet the criteria for the clinical diagnosis of acute bacterial rhinosinusitis and for those with severe rhinosinusitis symptoms-especially those with unilateral facial pain-regardless of duration of illness. For initial treatment, the most narrow-spectrum agent active against the likely pathogens, Streptococcus pneumoniae and Haemophilus influenzae, should be used. PMID- 11385343 TI - Principles of appropriate antibiotic use for treatment of nonspecific upper respiratory tract infections in adults: background. AB - The following principles of appropriate antibiotic use for adults with nonspecific upper respiratory tract infections apply to immunocompetent adults without complicating comorbid conditions, such as chronic lung or heart disease. 1. The diagnosis of nonspecific upper respiratory tract infection or acute rhinopharyngitis should be used to denote an acute infection that is typically viral in origin and in which sinus, pharyngeal, and lower airway symptoms, although frequently present, are not prominent. 2. Antibiotic treatment of adults with nonspecific upper respiratory tract infection does not enhance illness resolution and is not recommended. Studies specifically testing the impact of antibiotic treatment on complications of nonspecific upper respiratory tract infections have not been performed in adults. Life-threatening complications of upper respiratory tract infection are rare. 3. Purulent secretions from the nares or throat (commonly observed in patients with uncomplicated upper respiratory tract infection) predict neither bacterial infection nor benefit from antibiotic treatment. PMID- 11385345 TI - Principles of appropriate antibiotic use for acute pharyngitis in adults: background. AB - The following principles of appropriate antibiotic use for adults with acute pharyngitis apply to immunocompetent adults without complicated comorbid conditions, such as chronic lung or heart disease, and history of rheumatic fever. They do not apply during known outbreaks of group A streptococcus. 1. Group A beta-hemolytic streptococcus (GABHS) is the causal agent in approximately 10% of adult cases of pharyngitis. The large majority of adults with acute pharyngitis have a self-limited illness, for which supportive care only is needed. 2. Antibiotic treatment of adult pharyngitis benefits only those patients with GABHS infection. All patients with pharyngitis should be offered appropriate doses of analgesics and antipyretics, as well as other supportive care. 3. Limit antibiotic prescriptions to patients who are most likely to have GABHS infection. Clinically screen all adult patients with pharyngitis for the presence of the four Centor criteria: history of fever, tonsillar exudates, no cough, and tender anterior cervical lymphadenopathy (lymphadenitis). Do not test or treat patients with none or only one of these criteria, since these patients are unlikely to have GABHS infection. For patients with two or more criteria the following strategies are appropriate: (a) Test patients with two, three, or four criteria by using a rapid antigen test, and limit antibiotic therapy to patients with positive test results; (b) test patients with two or three criteria by using a rapid antigen test, and limit antibiotic therapy to patients with positive test results or patients with four criteria; or (c) do not use any diagnostic tests, and limit antibiotic therapy to patients with three or four criteria. 4. Throat cultures are not recommended for the routine primary evaluation of adults with pharyngitis or for confirmation of negative results on rapid antigen tests when the test sensitivity exceeds 80%. Throat cultures may be indicated as part of investigations of outbreaks of GABHS disease, for monitoring the development and spread of antibiotic resistance, or when such pathogens as gonococcus are being considered. 5. The preferred antibiotic for treatment of acute GABHS pharyngitis is penicillin, or erythromycin in a penicillin-allergic patient. PMID- 11385346 TI - Principles of appropriate antibiotic use for treatment of uncomplicated acute bronchitis: background. AB - The following principles of appropriate antibiotic use for adults with acute bronchitis apply to immunocompetent adults without complicating comorbid conditions, such as chronic lung or heart disease. The evaluation of adults with an acute cough illness or a presumptive diagnosis of uncomplicated acute bronchitis should focus on ruling out serious illness, particularly pneumonia. In healthy, nonelderly adults, pneumonia is uncommon in the absence of vital sign abnormalities or asymmetrical lung sounds, and chest radiography is usually not indicated. In patients with cough lasting 3 weeks or longer, chest radiography may be warranted in the absence of other known causes. Routine antibiotic treatment of uncomplicated acute bronchitis is not recommended, regardless of duration of cough. If pertussis infection is suspected (an unusual circumstance), a diagnostic test should be performed and antimicrobial therapy initiated. Patient satisfaction with care for acute bronchitis depends most on physician- patient communication rather than on antibiotic treatment. PMID- 11385347 TI - Fatality reduction by safety belts for front-seat occupants of cars and light trucks. PMID- 11385348 TI - Commentary: Traffic safety--an American export. PMID- 11385350 TI - Survey of international search and rescue teams after the Ji Ji earthquake in Taiwan. PMID- 11385351 TI - Intravenous calcium gluconate in the treatment of hydrofluoric acid burns. PMID- 11385352 TI - Comparison of class III antiarrhythmic drugs versus digoxin for the reversion of new-onset atrial fibrillation. PMID- 11385354 TI - Emergency removal of hard metal or ceramic finger rings. PMID- 11385356 TI - Clinical policy: critical issues in the evaluation and management of patients presenting with syncope. PMID- 11385357 TI - Reverse transcription-polymerase chain reaction quantitation of steroid receptor mRNA levels. PMID- 11385358 TI - Thrombogenesis or thrombogenic risk? PMID- 11385359 TI - Quantitative measurements of steroid receptors and their messenger ribonucleic acids with a special emphasis on polymerase chain reaction. PMID- 11385360 TI - Thrombogenesis in sickle cell disease. AB - Thirty-three subjects with sickle cell disease (SCD), 11 during episodes of pain and 22 during periods without pain, were evaluated for in vivo thrombogenic activities as compared with 10 normal black control subjects. Measurements were performed for (1) platelet surface activation, assessing flow cytometric expression of activated integrin alpha(IIb)beta(3) receptor (GPIIb/IIIa, CD41a) and P-selectin (CD62p); (2) platelet and erythrocyte surface procoagulant activities, measuring flow cytometric binding of activated factor (FVa) and annexin V; (3) plasma levels of platelet-specific secreted proteins platelet factor 4 (PF4) and beta-thromboglobulin (betaTG); (4) plasma markers of thrombin generation, prothrombin activation fragment (F(1.2)), and thrombin: antithrombin complex (TAT); and (5) plasma markers of fibrinolysis, D -dimer, and plasmin:antiplasmin complex (PAP). As compared with control subjects, asymptomatic subjects with SCD demonstrated significantly increased platelet activation (P <.01 for P-selectin and annexin V binding), elevated plasma levels of PF4 and betaTG (P <.01 and P <.03, respectively), and increased plasma concentrations of F(1.2), TAT, PAP, and D -dimer (P <.05 in all cases). During episodes of SCD pain, platelet activation was increased as compared with periods without pain (P <.01 for expression of activated integrin alpha(IIb)beta(3) receptor and P-selectin and binding of FVa and annexin V), erythrocytes expressed procoagulant activities (P <.01 for FVa and annexin V binding), and platelet microparticles appeared in the circulation (3% to 30%; P <.001). SCD pain episodes were associated with elevated plasma levels of F(1.2), TAT, PAP, and D dimer (P <.05 as compared with asymptomatic intervals). The frequency of pain episodes correlated with enhanced platelet procoagulant activity (r = 0.61, P <.05) and elevated plasma fibrinolytic activity (r = 0.74, P <.01) measured during periods without pain. Plasma fibrinolytic activity was inversely correlated with time to the next pain episode (r = -0.50, P <.05). Thus, asymptomatic subjects with SCD exhibit ongoing platelet activation, thrombin generation, and fibrinolysis that increases during episodes of pain. These changes are predictive of frequency of pain and interval to next pain episode, thereby implicating thrombogenic activity in the development of SCD pain episodes. PMID- 11385361 TI - Laboratory diagnosis of heparin-induced thrombocytopenia type II after clearance of platelet factor 4/heparin complex. AB - Laboratory confirmation of heparin-induced thrombocytopenia (HIT) is limited by assay sensitivity. We investigated whether laboratory confirmation can be improved after antigen clearance by determining free antibody and combining the results of antigenic and biologic assays. Blood samples were collected over 5 to 6 weeks in 14 HIT patients. As an antigenic assay, the fluorescence-linked immunofiltration assay (FLIFA) was performed, and as a biologic assay, the carbon 14-labeled serotonin release assay was performed. At day 1 when heparin was stopped, 11 of 14 patients showed positive results in both assays; thus each assay had a sensitivity of 80%. The 3 patients with negative results seroconverted in one or both assays during the subsequent 7 days. Combining the positive results of the assays increased the sensitivity to 100% at day 7, regardless of whether the antigenic or the biologic assay was performed first. Both assays became negative in all patients within 5 to 6 weeks. The sensitivity of antigen and biologic assays in HIT patients increased to 100% after the time course of the heparin-induced antibody. We assume that in some HIT patients the free antibody can be detected after withdrawal of heparin and after clearance of the platelet-factor 4/heparin complex. PMID- 11385362 TI - Effect of gemfibrozil on the composition and oxidation properties of very-low density lipoprotein and high-density lipoprotein in patients with hypertriglyceridemia. AB - Recent studies suggest that both oxidized very-low-density lipoprotein (VLDL) and oxidized high-density lipoprotein (HDL) may play a role in the pathogenesis of atherosclerosis. Gemfibrozil is widely used and is reported to decrease VLDL levels and increase HDL levels. The aim of this study was to investigate the effect of gemfibrozil on the chemical composition and oxidative susceptibility of VLDL and HDL and their relationship with atherosclerosis. Twenty patients with hypertriglyceridemia were treated with 300 mg gemfibrozil, 3 times daily, for 12 weeks. Venous blood samples were collected before treatment, at the end of treatment, and 4 weeks after the end of treatment. Gemfibrozil effectively lowered concentrations of plasma lipid, apolipoprotein (apo) B, and apo E. The lipid and protein content of VLDL were also decreased, but not by the same extent. The surface-to-core ratio and apo E/apo B ratio of VLDL particles were increased after gemfibrozil treatment. HDL(2) cholesteryl ester and HDL(3) apo A II content were also increased. Gemfibrozil treatment lowered levels of lipid peroxides in both VLDL and HDL particles. The susceptibility of VLDL to oxidation was unchanged, whereas maximal peroxide production was decreased. The oxidative susceptibility of both HDL(2) and HDL(3) decreased with gemfibrozil treatment. These results indicate that after gemfibrozil treatment, VLDL and HDL particles in patients with hypertriglyceridemia are less atherogenic, which may explain why gemfibrozil treatment is beneficial in terms of coronary heart disease in hypertriglyceridemia. PMID- 11385363 TI - Reverse transcriptase template switching during reverse transcriptase-polymerase chain reaction: artificial generation of deletions in ribonucleotide reductase mRNA. AB - Using reverse transcriptase-polymerase chain reaction (RT-PCR), we have recently described a bona fide deletion within the coding sequence of the large subunit of ribonucleotide reductase (R1) mRNA in colon cancer. Consecutive studies have raised questions about the nature of this phenomenon, because the corresponding genomic alteration at the DNA level or an aberrant protein could not be detected. Thus we considered an in vitro artifact during RT-PCR as a possible explanation for this observation. In contrast to reverse transcriptase, Taq DNA polymerase or C. therm DNA polymerase did not generate the aberrant product, suggesting the demand for the template switching activity intrinsic to retroviral reverse transcriptases. In fact, virtually the same deletion was observed in RT-PCR experiments when in vitro transcribed R1 mRNA was used. Considering structural prerequisites for template switching within R1 mRNA, we show that two direct repeats adjacent to a strong stem-loop secondary structure flank the deleted region of 1851 base pairs. Because several mRNAs encoding proteins of clinical and diagnostic importance fulfill these criteria, template switching enhances the potential risk of observing artifacts when interpreting results from RT-PCR studies. As shown in the present example, this may involve the artificial generation and the misinterpretation of PCR fragments amplified from targets relevant to tumor biology or cancer pharmacology. As a possible solution, one step PCR with C. therm polymerase should be considered. This polymerase eliminates the artificial generation of aberrant mRNA signals observed during cDNA synthesis. PMID- 11385364 TI - Pneumocystis carinii pneumonia alters expression and distribution of lung collectins SP-A and SP-D. AB - Surfactant proteins SP-A and SP-D, members of the collectin family, have been shown to play a significant role in lung host defense. Both proteins selectively bind Pneumocystis carinii (PC) organisms and modulate the interaction of this pathogen with alveolar macrophages. We hypothesized that the expression and distribution of lung collectins SP-A and SP-D is altered by PC lung infection. PC organisms (2 x 10(5)) were inoculated intratracheally into C.B-17 scid/scid mice that do not require steroids for immunosuppression. Four weeks after inoculation, bronchoalveolar lavage (BAL) fluid was fractionated into three fractions-cell pellet, large aggregate (LA), and small aggregate (SA) surfactant-and each fraction was analyzed for the expression of surfactant components. In uninfected mice, the majority of SP-A (62% +/- 10%) was found in association with lipids in the LA fraction, while 55% +/- 14% of SP-D was distributed in the SA fraction. In contrast, both hydrophobic proteins SP-B and SP-C were associated exclusively with LA. PC infection resulted in major changes in the expression of all surfactant components. Total protein content of LA was unchanged by PC infection (115% +/- 18% of control), whereas SA protein content markedly increased (240% +/ 18% of control level, P <.001). In contrast, the phospholipid content of LA was significantly decreased (53% +/- 5% of control level, P <.001), whereas the SA phospholipid content of infected mice was increased (172% +/- 16% of control level, P <.001). By Western blotting, PC pneumonia (PCP) induced a 3-fold increase in the total alveolar SP-D protein that was reflected mainly in increases in SA SP-D (454% +/- 135% of control, P <.05). The total alveolar SP-A protein content was also increased in PCP because of a large increase in SP-A in SA (720% +/- 115% of control, P <.05); SP-A levels in LA were unchanged. The increases in lung collectin expression were selective, because PCP resulted in the down-regulation of both SP-B and SP-C in LA (5% +/- 2% and 13% +/- 2% of control, respectively, P <.001). We conclude that PCP induces marked elevations in alveolar collectin levels because of increased expression and accumulation of SP-A and SP-D protein in SA surfactant. PMID- 11385365 TI - Consensus statement on submission and publication of manuscripts. PMID- 11385366 TI - Should screening for lung cancer be revisited? PMID- 11385367 TI - Pulmonary vascular resistance after cardiopulmonary bypass in infants: effect on postoperative recovery. AB - OBJECTIVE: We sought to define the contemporary clinical effect of increased pulmonary vascular resistance in infants after congenital heart operations with cardiopulmonary bypass. METHODS: Fifteen infants (median age, 0.31 years; median weight, 5.1 kg) underwent cardiac operations involving cardiopulmonary bypass (range, 49-147 minutes). Pulmonary vascular resistance was measured in the immediate postoperative period in the intensive care unit by means of the direct Fick principle, with respiratory mass spectrometry to measure oxygen consumption. The effect of ventilation with an inspired oxygen fraction of 0.65, with additional infusion of L -arginine, substance P, and inhaled nitric oxide, was assessed and subsequently correlated with the length of mechanical ventilation from the end of cardiopulmonary bypass to successful extubation. RESULTS: Overall, pulmonary vascular resistance at baseline (11.7 +/- 5.6 WU. m(2)) could be reduced to a minimum of 6.1 +/- 3.5 WU. m(2). The ventilatory time was 0.86 to 14.9 days (median, 1.75 days) and correlated directly with the lowest pulmonary vascular resistance value achieved during the pulmonary vascular resistance study (r (2) = 0.64, P <.01). The patient subgroup with mechanical ventilation of greater than 2 days had significantly higher pulmonary vascular resistance at all stages of the study protocol, and in this group there was a correlation of cardiopulmonary bypass time and ventilatory support time (r (2) = 0.48, P <.05). CONCLUSION: Increased pulmonary vascular resistance, either directly or as a surrogate of the systemic inflammatory response after cardiopulmonary bypass, continues to have a significant effect on postoperative recovery of infants after cardiac operations. PMID- 11385368 TI - A case for anatomic correction in atrioventricular discordance? Effects of surgery on tricuspid valve function. AB - OBJECTIVE: To assess tricuspid valve function in atrioventricular discordance after palliative procedures (pulmonary artery banding and Blalock-Taussig shunt) and corrective procedures (anatomic and physiologic repair). METHODS: Tricuspid valve dysfunction was assessed by transthoracic echocardiography and graded as no regurgitation (0), mild (1), moderate (2), and severe (3) before and after palliative and corrective procedures performed in 97 patients with atrioventricular discordance between 1988 and 1999. Thirty-two percent had an isolated ventricular septal defect, 43% had a ventricular septal defect and pulmonary stenosis, and 16% had pulmonary stenosis. Twenty-six patients underwent pulmonary artery banding and 28 had a Blalock-Taussig shunt. Seventy patients underwent physiologic and 19 underwent anatomic repair. Six patients underwent one-ventricle repair. RESULTS: After pulmonary artery banding, the tricuspid regurgitation score decreased from 1.7 +/- 0.8 to 0.9 +/- 0.6 (P <.001). In patients who underwent a Blalock-Taussig shunt, the tricuspid regurgitation score increased from 0.7 +/- 0.5 preoperatively to 1.4 +/- 0.6 postoperatively (P <.001). After physiologic repair, there was no significant change in the tricuspid regurgitation score; however, 7 patients required additional repair or replacement. The regurgitation score was significantly reduced from 1.5 +/- 0.8 to 0.4 +/- 0.5 (P <.001) after anatomic repair. The operative mortality in patients who underwent physiologic repair was 7% as compared with 0% in the anatomic repair group (P =.59). The median follow-up was 3.2 years. CONCLUSIONS: Right ventricular volume loading (shunt) worsens tricuspid regurgitation, whereas volume reduction (banding) or left-to-right septal shift (anatomic repair) has beneficial effects. We have not observed a significant change in the tricuspid regurgitation score after physiologic repair. Anatomic repair can be performed in selected patients with atrioventricular discordance and provides superior functional results. PMID- 11385369 TI - The anatomy of the septal perforating arteries in normal and congenitally malformed hearts. AB - BACKGROUND: Many cardiac operations involve incisions and sutures on or near the ventricular septum. These jeopardize the septal perforating arteries. Our aim was to provide guidelines for the surgeon to predict the site of these vessels. METHODS AND RESULTS: We dissected 50 hearts. In 16 of these we also conducted histologic examination of the area of the septum containing the atrioventricular node, the penetrating bundle (of His), and the branching atrioventricular bundle to elucidate the source of the vascular supply to these structures. The major perforating septal arteries arise from the superior interventricular artery or, in hearts with a rudimentary right ventricle, from the superior delimiting artery. The first is usually the largest. The location of this artery can be predicted relative to the position of the medial papillary muscle. In abnormal hearts, holes within the ventricular septum in the presence of a well-developed muscular outlet septum were found to deviate the path of the septal perforating arteries in a predictable manner. The triangular area bordered by the margin of the ventricular septal defect, the muscular outlet septum, and the medial papillary muscle is free of major perforating arteries. The histologic studies showed that the conduction tissues at the base of the ventricles tend to receive their blood supply from arteries arising from the inferior interventricular artery, except in double-inlet left ventricle, in which the arterial supply is from the right-sided delimiting artery. CONCLUSION: The location of the first superior septal perforating artery is predictable in many cases. Its course leaves a triangular area on the muscular ventricular septum that is free of major arteries. PMID- 11385370 TI - Detection of early-stage lung cancer: computed tomographic scan or chest radiograph? AB - OBJECTIVE: Computed tomography has recently been proposed as a useful method for the early detection of lung cancer. In this study we compared the stage distribution of lung cancers detected by a computed tomographic scan with that of lung cancers detected by a routine chest x-ray film. METHODS: Two groups of patients with biopsy-proven non-small cell lung cancer were reviewed. In the first group of 32 patients, the tumors were detected by a computed tomographic scan. In a second group (n = 101), the lung cancers were detected on routine chest x-ray films. Patients with pulmonary symptoms or a history of cancer were excluded. RESULTS: There was no difference in age, sex, or cell-type distribution between the 2 groups. A significantly greater number of patients undergoing a computed tomographic scan had stage IA disease compared with those having an x ray film. Of the 32 patients in the group having a scan, 10 had tumors 1 cm or less in size versus 6 of 101 in the group having a chest radiograph. Additionally, there was a significant reduction in advanced stage disease in the group having a scan. CONCLUSIONS: In this retrospective study, a higher incidence of stage IA lung cancers and significantly fewer cases of more advanced disease were observed in patients screened with computed tomography than in those having a chest radiograph. These data suggest that computed tomographic screening may be of value in improving the survival of patients with non-small cell lung cancer. PMID- 11385371 TI - The prognostic value of natural killer cell infiltration in resected pulmonary adenocarcinoma. AB - OBJECTIVE: Natural cytotoxicity caused by mediated natural killer cells is believed to play an important role in host-cancer defense mechanisms. Immunohistochemically, we have detected natural killer cells in tissue specimens from patients with pulmonary adenocarcinoma and have assessed their clinical characteristics. METHODS: Using the monoclonal antibody for CD57 specific marker for natural killer cells, we quantified natural killer cell infiltration in 150 patients with pulmonary adenocarcinoma who underwent curative tumor resection to investigate the relationship between natural killer cell counts and clinicopathologic factors and prognosis. RESULTS: The natural killer cell count was significantly related to the regulation of tumor progression, involving T classification, N classification, and stage (P =.01 for T classification or stage; P =.02 for N classification). A significant difference in the rate of patient survival was detected between those patients whose tumors had either high or low natural killer cell counts in both the overall and stage I groups (P =.0002 for the overall group; P =.049 for the stage I group). CONCLUSION: These data indicate that natural killer infiltration may contribute to the regulation of tumor progression and that the natural killer cell count can serve as a useful prognostic marker in overall and stage I pulmonary adenocarcinoma. PMID- 11385372 TI - Preoperative evaluation of cardiopulmonary reserve with the use of expired gas analysis during exercise testing in patients with squamous cell carcinoma of the thoracic esophagus. AB - OBJECTIVE: We evaluated the usefulness of analyzing expired gas during exercise testing for the prediction of postoperative cardiopulmonary complications in patients with esophageal carcinoma. BACKGROUND DATA: Radical esophagectomy with 3 field lymphadenectomy is performed in patients with thoracic esophageal carcinoma but has a high risk of postoperative complications. To reduce the surgical risk, we performed preoperative risk analysis using 8 factors. Although hospital mortality was decreased when this risk analysis was used, severe cardiopulmonary complications still occurred. METHODS: The study group consisted of 91 patients who had undergone curative esophagectomy with 3-field lymphadenectomy. The maximum oxygen uptake, anaerobic threshold, vital capacity, percent vital capacity, forced expiratory volume in 1 second, percent forced expiratory volume, V.(25)/HT, forced expired flow at 75% of forced vital capacity to height ratio (FEF(75%)/HT), forced expired flow at 50% to 75% of forced vital capacity ratio (FEF(50%)/FEF(75%)), percent diffusion capacity for carbon monoxide, and arterial oxygen tension were measured. Patients were divided into 2 groups on the basis of the presence or absence of postoperative cardiopulmonary complications. RESULTS: Only the maximum oxygen uptake was significantly different between the 2 groups. All patients were grouped according to the value of the maximum oxygen uptake, and the occurrence of postoperative cardiopulmonary complications was calculated for each group. A cardiopulmonary complication rate of 86% was found for patients with a maximum oxygen uptake of less than 699 mL. min(-1). m(-2); for those with a value of 700 to 799 mL. min(-1). m(-2), the complication rate was 44%. CONCLUSIONS: The maximum oxygen uptake obtained by expired gas analysis during exercise testing correlates with the postoperative cardiopulmonary complication rate. On the basis of these results, esophagectomy with 3-field lymphadenectomy can be safely performed in patients with a maximum oxygen uptake of at least 800 mL. min(-1). m(-2). PMID- 11385373 TI - Lung transplant reperfusion injury involves pulmonary macrophages and circulating leukocytes in a biphasic response. AB - OBJECTIVE: Both donor pulmonary macrophages and recipient circulating leukocytes may be involved in reperfusion injury after lung transplantation. By using the macrophage inhibitor gadolinium chloride and leukocyte filters, we attempted to identify the roles of these two populations of cells in lung transplant reperfusion injury. METHODS: With our isolated, ventilated, blood-perfused rabbit lung model, all groups underwent lung harvest followed by 18-hour cold storage and 2-hour blood reperfusion. Measurements of pulmonary artery pressure, lung compliance, and arterial oxygenation were obtained. Group I (n = 8) served as a control. Group II (n = 8) received gadolinium chloride at 14 mg/kg 24 hours before lung harvest. Group III (n = 8) received leukocyte-depleted blood reperfusion by means of a leukocyte filter. RESULTS: The gadolinium chloride group had significantly improved arterial oxygenation and pulmonary artery pressure measurements compared with control subjects and an improved arterial oxygenation compared with the filter group after 30 minutes of reperfusion. After 120 minutes of reperfusion, however, the filter group had significantly improved arterial oxygenation and pulmonary artery pressure measurements compared with the control group and an improved arterial oxygenation compared with the gadolinium chloride group. CONCLUSIONS: Lung transplant reperfusion injury occurs in two phases. The early phase is mediated by donor pulmonary macrophages and is followed by a late injury induced by recipient circulating leukocytes. PMID- 11385374 TI - Pneumonectomy for malignant disease: factors affecting early morbidity and mortality. AB - OBJECTIVE: The purpose of this report is to analyze factors affecting morbidity and mortality after pneumonectomy for malignant disease. METHODS: We retrospectively reviewed the cases of all patients who underwent pneumonectomy for malignancy at the Mayo Clinic. Between January 1, 1985, and September 30, 1998, 639 patients (469 men and 170 women) were identified. Median age was 64 years (range 20 to 86 years). Indication for pneumonectomy was primary lung cancer in 607 (95.0%) patients and metastatic disease in 32 (5.0%). Factors affecting morbidity and mortality were analyzed by univariate and multivariate analysis. RESULTS: Cardiopulmonary complications occurred in 245 patients (38.3%; 95% confidence interval 34.6%-42.2%). Factors adversely affecting morbidity with univariate analysis included age (P <.0001), male sex (P =.04), associated respiratory (P =.02) or cardiovascular disease (P <.0001), cigarette smoking (P =.02), decreased vital capacity (P =.01), forced expiratory volume in 1 second (P <.0001), forced vital capacity (P =.002), diffusion capacity of the lung to carbon monoxide (P =.005), oxygen saturation (P <.05), arterial PO (2) (P =.007), preoperative radiation (P =.02), bronchial stump reinforcement (P =.007), crystalloid infusion (P =.01), and blood transfusion (P =.02). Factors adversely affecting morbidity with multivariate analysis included age (P =.0001), associated cardiovascular disease (P =.001), and bronchial stump reinforcement (P =.0005). There were 45 deaths (7.0%; 95% confidence intervals 5.2%-9.3%). Factors adversely affecting mortality with univariate analysis included associated cardiovascular (P <.0001) or hematologic disease (P <.005), lower preoperative serum hemoglobin level (P =.004), preoperative chemotherapy (P =.01), decreased diffusion capacity of lung to carbon monoxide (P =.002), right pneumonectomy (P =.0006), extended resection (P =.04), bronchial stump reinforcement (P =.007), and crystalloid infusion (P =.01). Factors affecting mortality with multivariate analysis included hematologic disease (P =.01), lower preoperative serum hemoglobin (P =.003), and completion pneumonectomy (P =.01). CONCLUSION: Multiple factors adversely affected morbidity and mortality after pneumonectomy for malignant disease. Appropriate selection and meticulous perioperative care are paramount to minimize risks in those patients who require pneumonectomy. PMID- 11385375 TI - Coronary artery bypass grafting in non-dialysis-dependent mild-to-moderate renal dysfunction. AB - OBJECTIVES: The effect of mild-to-moderate elevation of preoperative serum creatinine levels on morbidity and mortality from coronary artery bypass grafting has not been investigated in a large multivariable model incorporating preoperative and intraoperative variables. Our first objective was to ascertain the effect of a mild-to-moderate elevation in the preoperative serum creatinine level on the need for mechanical renal support; the duration of special care and total postoperative stay; the occurrence of infective, respiratory, and neurologic complications; and hospital mortality. Our second objective was to ascertain which patient variables contributed to an increase in the serum creatinine level in association with coronary artery bypass grafting. METHODS: A total of 1427 patients who had no known pre-existing renal disease and who were undergoing first-time coronary artery bypass grafting with cardiopulmonary bypass were recruited for the study. Patients were divided, on the basis of preoperative serum creatinine level, into 3 groups as follows: creatinine level of less than 130 micromol. L(-1); creatinine level of 130 to 149 micromol. L(-1); and creatinine level of 150 micromol. L(-1) or greater. A multivariable stepwise logistic regression analysis was used, and variables significant at the 5% level were included when developing the final multivariable models. RESULTS: Multivariable analysis showed that elevation of the preoperative serum creatinine level to 130 micromol. L(-1) or greater increased the likelihood of needing mechanical renal support postoperatively (P <.001), as well as the need for postoperative special care (P <.001) and total hospital stay (P <.001). In hospital mortality was also significantly elevated as the preoperative creatinine level rose to 130 to 149 micromol. L(-1) (P =.045) and to 150 micromol. L(-1) or greater (P <.001). It was further observed that patients with preoperative serum creatinine levels of 130 to 149 micromol. L(-1) (P =.02), patients with preoperative serum creatinine levels of 150 micromol. L(-1) or greater (P =.001), hypertensive patients (P =.007), patients with angina of New York Heart Association class III or greater (P =.001), patients having a nonelective operation (P =.002), and patients having a prolonged cardiopulmonary bypass time (P =.008) had a significantly greater increase in the serum creatinine level as a result of coronary artery bypass grafting. Of particular note was the finding that the method of myocardial protection (cardioplegia or crossclamp fibrillation) did not significantly influence in-hospital mortality, need for mechanical renal support, or special care or total postoperative hospital stay. CONCLUSIONS: A mild elevation (130-149 micromol. L(-1)) in the preoperative serum creatinine level significantly increases the need for mechanical renal support, the duration of special care and total postoperative stay, and the in-hospital mortality. As the preoperative serum creatinine level increases further (> or =150 micromol. L(-1)), this effect is more pronounced. No significant difference in outcome was observed between the use of cardioplegia or crossclamp fibrillation for myocardial protection. PMID- 11385376 TI - Twenty years' experience with the Medtronic Hall valve. AB - OBJECTIVE: To assess the performance of the Medtronic Hall valve (Medtronic, Inc, Minneapolis, Minn) in one institution over a 20-year period. METHODS: Since 1979, Medtronic Hall valves have been used in 1766 procedures (736 aortic, 796 mitral, and 234 double). Patients were followed up prospectively at 6- to 12-month intervals for a total of 12,688 follow-up years. Anticoagulation data (international normalized ratio) were recorded for all patients (approximately 95,000 observations). RESULTS: Linearized rates of valve-related late death for aortic, mitral, and double valve replacement were 0.8%/y, 0.9%/y, and 1.1%/y, respectively. Risk factors for late mortality were (relative risk) diabetes (1.9), decade of age (1.6), concomitant coronary artery bypass grafting (1.4), hypertension (1.3), non-sinus rhythm (1.3), large valve size (1.1), valve regurgitation (1.3), and male sex (1.2). For aortic, mitral, and double valve replacement, linearized rates (percent per year) of adverse events were valve thrombosis 0.04, 0.03, and 0.0; all thromboembolism 2.3, 4.0, and 3.4; stroke 0.6, 0.8, and 0.6; major hemorrhage 1.2, 1.4, and 1.6; and prosthetic endocarditis 0.4, 0.4, and 0.7. Risk factors for thromboembolism were (relative risk) mitral valve replacement (1.9), diabetes (1.8), hypertension (1.5), and history of embolism (1.4). CONCLUSION: At 20 years the Medtronic Hall valve demonstrates excellent durability, good hemodynamic performance, and very low thrombogenicity, with a valve thrombosis rate lower than those reported for bileaflet designs. With this prosthesis, both survival and thromboembolic events are predominantly determined by patient risk factors. PMID- 11385377 TI - Doppler microembolic signals in patients with two different types of bileaflet valves. AB - OBJECTIVES: This study was performed to evaluate the prevalence and counts of Doppler microembolic signals in patients with St Jude Medical valves (St Jude Medical, Inc, St Paul, Minn) and patients with ATS valves (ATS Medical, Inc, Minneapolis, Minn) and their relation to clinical parameters. METHODS: A total of 179 outpatients of the department of cardiothoracic surgery were examined. They included 98 men and 81 women, aged 61 +/- 11 years, with ATS (n = 91) or St Jude Medical (n = 88) valves in the aortic (n = 110), mitral (n = 39), or both positions (n = 30). Neurologic examination was followed by transcranial Doppler monitoring for microembolic signals. Monitoring was performed bilaterally over the middle cerebral arteries for 1 hour per session. RESULTS: Microembolic signal counts and prevalence were significantly higher in patients with St Jude Medical as compared with ATS valves. Valve type and presence of diabetes mellitus were the only predictors of microembolic signal prevalence on multivariate analysis. No influence of microembolic signals on cerebral embolic complications was established. Additionally, patients with a postoperative history of cerebral embolic complications did not have a higher number of microembolic signals than remaining patients. Interobserver variability was satisfactory. CONCLUSIONS: Patients with St Jude Medical valves were shown to have significantly higher microembolic signal counts than patients with ATS valves. However, our results suggest that microembolic signal counts cannot be used to predict cerebral embolic complications. Their relation to neuropsychologic deficits remains to be evaluated. PMID- 11385378 TI - Neurologic outcome after ascending aorta-aortic arch operations: effect of brain protection technique in high-risk patients. AB - OBJECTIVE: We sought to assess the optimal strategy for avoiding neurologic injury after aortic operations requiring hypothermic circulatory arrest. METHODS: All 717 patients who survived ascending aorta-aortic arch operations through a median sternotomy since 1986 were examined for factors influencing stroke. Temporary neurologic dysfunction was assessed in all patients who survived the operation without stroke since 1993. Multivariate analyses were carried out to determine independent risk factors for neurologic injury. RESULTS: Independent risk factors for stroke were as follows: age greater than 60 years (P <.001; odds ratio, 4.5); emergency operation (P =.02; odds ratio, 2.2); new preoperative neurologic symptoms (P =.05; odds ratio, 2.9); presence of clot or atheroma (P <.001; odds ratio, 4.4); mitral valve replacement or other concomitant procedures (P =.055; odds ratio, = 3.7); and total cerebral protection time, defined as the sum of hypothermic circulatory arrest and any retrograde or antegrade cerebral perfusion (P =.001; odds ratio, 1.02/min). In 453 patients surviving operations without stroke after 1993, independent risk factors for temporary neurologic dysfunction included age (P <.001; odds ratio, 1.06/y), dissection (P =.001; odds ratio, 2.2), need for coronary artery bypass grafting (P =.006; odds ratio, 2.1) or other procedures (P =.023; odds ratio, 3.4), and total cerebral protection time (P <.001; odds ratio, 1.02/min). When all patients with total cerebral protection times between 40 and 80 minutes were examined, the method of cerebral protection did not influence the occurrence of stroke, but antegrade cerebral perfusion resulted in a significant reduction in incidence on temporary neurologic dysfunction (P =.05; odds ratio, 0.3). CONCLUSIONS: The occurrence of stroke is principally determined by patient- and disease-related factors, but use of antegrade cerebral perfusion can significantly reduce the occurrence of temporary neurologic dysfunction. PMID- 11385379 TI - Closure of atrial septal defects without cardiopulmonary bypass: the sandwich operation. AB - BACKGROUND: Cardiopulmonary bypass has adverse effects on patient physiology. A prospective randomized trial was undertaken to evaluate closure of atrial septal defects with or without cardiopulmonary bypass. METHODS: Between August 1997 and March 2000, 150 patients with ostium secundum atrial septal defects were enrolled. Patients were randomized for repair without cardiopulmonary bypass (ie, the sandwich operation; n = 74) as a study group or with cardiopulmonary bypass (n = 76) as a control group. In the sandwich group the sandwich patch was passed into the right atrium and placed at the defect during transesophageal echocardiography. The patch was secured with external transfixing sutures and endoscopic staples. Clinical outcomes were compared. Outcome variables included perioperative morbidity, mortality, length of stay in the intensive care unit, hospital length of stay from operation to discharge, residual shunt, reoperation, hematologic profile, transfusion requirement, and hospital fee. The follow-up duration ranged from 1 to 29 months (mean, 8.7 +/- 7.5 months). RESULTS: There was no operative mortality. The success rate of the sandwich operation was 68 (92%) of 74 patients. Patients in the sandwich group had shorter operative times and lengths of stay in the intensive care unit, fewer requirements of transfusion, and lower hospital fees. The differences in postoperative drainage, arrhythmias, and hematologic profiles could be due to chance. Two patients in the sandwich group required reoperation for residual shunt. The risk factor for residual shunt was earlier date of repair. Mitral and tricuspid valve function was not adversely affected by the operation. CONCLUSIONS: Closure of atrial septal defects without cardiopulmonary bypass can be done effectively. Adverse effects of cardiopulmonary bypass can be avoided, as shown by improvements of postoperative parameters with the sandwich operation. PMID- 11385380 TI - Cold cardioplegic arrest enhances heat shock protein 70 in the heat-shocked rat heart. AB - BACKGROUND: Myocardial content of the 70-kd heat shock protein has been found to correlate with improved cardiac recovery after ischemia, but the mechanisms and conditions that regulate its level, particularly under clinical conditions, are unclear. The aim of this study was to assess the effect of hypothermic cardioplegic arrest and reperfusion on the expression of 70-kd heat shock protein in a protocol mimicking conditions of preservation for cardiac transplantation. METHODS: Heat-shocked and control hearts were subjected to 4 hours of cardioplegic arrest and global ischemia at 4 degrees C and then to 20 minutes of reperfusion. Hearts were freeze clamped at different time points-after 15 minutes of Langendorff perfusion, at the end of ischemia, and after 20 minutes of reperfusion, and analyzed for heat shock protein 70 content by Western blotting. Another set of hearts was subjected to 10 minutes of normothermic ischemia and 20 minutes of reperfusion followed by freeze clamping and analysis of heat shock protein 70 content as in cardioplegic arrest protocol. Cardiac function was measured by means of a left ventricular balloon at the end of reperfusion. RESULTS: Preischemic concentration of 70-kd heat shock protein was increased in heat-shocked hearts compared with control hearts. The content of 70-kd heat shock protein in heat-shocked hearts was further increased from 5.0 +/- 2.4 ng/microg at the end of ischemia to 11.0 +/- 4.9 ng/microg (n = 8, mean +/- SD; P <.05) at 20 minutes of reperfusion after cold cardioplegic arrest. No further rise in 70 kd heat shock protein of the heat-shocked hearts was observed after normothermic ischemia. Maximal developed pressure was 120.8 +/- 13.4 mm Hg in control hearts compared with 164.7 +/- 22.5 mm Hg in heat-shocked hearts (n = 5, mean +/- SD; P =.037) after cardioplegic arrest. By contrast, after normothermic ischemia, maximum developed pressure was 111.2 +/- 10.9 mm Hg in control hearts compared with 139.2 +/- 11.0 mm Hg in heat-shocked hearts (n = 4, mean +/- SD; P =.031). CONCLUSION: Hypothermic cardioplegic arrest but not short normothermic ischemia triggered a further increase in the level of 70-kd heat shock protein in heat shocked rat hearts, which may enhance endogenous cardiac protection. PMID- 11385381 TI - Pulmonary metabolism of endothelin 1 during on-pump and beating heart coronary artery bypass operations. AB - BACKGROUND: Coronary artery bypass operations are associated with increased circulating levels of the powerful vasoconstrictor endothelin 1. The pulmonary circulation is an important site for both production and clearance of endothelin 1. Lung endothelial injury resulting from cardiopulmonary bypass could modify pulmonary endothelin 1 metabolism through an increase in production, a reduction in removal, or a combination of both. METHODS: Pulmonary endothelin 1 kinetics were quantified by using the indicator-dilution technique in patients undergoing coronary artery bypass grafting with (n = 11) or without cardiopulmonary bypass (ie, beating heart; n = 10). Mixed venous endothelin 1 levels were also measured in samples from the pulmonary artery, and systemic levels were obtained from the radial artery. RESULTS: Pulmonary artery endothelin 1 levels were similar before and after cardiopulmonary bypass, with means of 1.59 +/- 0.37 pg/mL and 1.33 +/- 0.15 pg/mL (P =.45), respectively. Systemic endothelin 1 levels, however, increased after bypass from 1.64 +/- 0.22 pg/mL to 2.07 +/- 0.16 pg/mL (P =.01). In the beating heart group, endothelin 1 levels before and after the operation were similar in the pulmonary artery (1.25 +/- 0.27 pg/mL and 1.45 +/- 0.31 pg/mL, respectively; P =.38), as well as in the radial artery (1.70 +/- 0.26 pg/mL and 1.73 +/- 0.35 pg/mL, respectively; P =.92). The capacity to clear endothelin 1 from the pulmonary circulation, as computed from the permeability surface area product for endothelin 1, was not affected by cardiopulmonary bypass before and after the operation (25.19 +/- 2.67 mL/s and 23.12 +/- 4.39 mL/s, respectively; P =.49). It was similar and also unaffected in the beating heart group. CONCLUSION: Coronary artery bypass grafting with cardiopulmonary bypass is associated with an increase in systemic endothelin 1 levels. The mechanism involved is not related to a decreased pulmonary clearance of endothelin 1 from the systemic circulation but rather to an increased endothelin 1 release by the lungs. PMID- 11385382 TI - Outlet strut fracture of Bjork-Shiley convexo-concave valves: can valve manufacturing characteristics explain the risk? AB - BACKGROUND: Bjork-Shiley 60 degrees convexo-concave prosthetic heart valves (Shiley, Inc, Irvine, Calif, a subsidiary of Pfizer, Inc) continue to be a concern for approximately 35,000 nonexplanted patients worldwide, with approximately 600 events reported to the manufacturer to date. Fractures of the outlet struts of the valves began to appear in the early 1980s and have continued to the present, but their causes are only partially understood. METHODS: A matched case-control study was conducted evaluating manufacturing records for 52 valves with outlet strut fractures and 248 control subjects matched for age at implantation, valve size, and valve position. RESULTS: In addition to the risk factors recognized as determinants of outlet strut fracture, the United Kingdom case-control study has observed 7- to 9-fold increased risk with performance of multiple hook deflection tests. This test was performed more than once, usually after rework on the valve. Six valves in this study underwent multiple hook deflection tests, of which 4 experienced an outlet strut fracture. Cracks and further rework were noted for these valves. Significant associations were also observed between outlet strut fracture and disc-to-strut gap measurements taken before the attachment of the sewing ring. CONCLUSIONS: It is our view that a combination of factors related to valve design, manufacturing process, and patient characteristics are responsible for outlet strut fractures of Bjork Shiley convexo-concave valves. Multiple hook deflection tests have emerged as a potential new risk factor for outlet strut fracture in both The Netherlands and the United Kingdom. This factor appears to be correlated with the presence of other abnormalities. A further study is needed to investigate the factors correlated with multiple hook deflection tests. On confirmation of risk, the presence of multiple hook deflection tests may be added to equations, quantifying the risk of outlet strut fracture for comparison against risk of mortality and serious morbidity from explant operations. PMID- 11385383 TI - Cerebral injury during cardiopulmonary bypass: emboli impair memory. AB - OBJECTIVES: Cognitive deficits occur in up to 80% of patients after cardiac surgery. We investigated the influence of cerebral perfusion and embolization during cardiopulmonary bypass on cognitive function and recovery. METHODS: Cerebrovascular reactivity was measured in 70 patients before coronary operations in which nonpulsatile bypass was used. Throughout the operations, middle cerebral artery flow velocity and embolization were recorded by transcranial Doppler and regional oxygen saturation was recorded by near-infrared spectroscopy. Cognitive function was measured by a computerized battery of tests before the operation and 1 week, 2 months, and 6 months after surgery. Elderly patients undergoing urologic surgery served as controls. RESULTS: Cerebrovascular reactivity was impaired preoperatively in 49 patients. Median (interquartile range) regional cerebral oxygen saturation fell during bypass by 10% (6%-15%), indicating increased oxygen extraction, whereas mean middle cerebral flow velocity increased significantly by a median of 6 cm/s (both P <.0001, Wilcoxon), suggesting increased arterial tone. More than 200 emboli were detected in 40 patients, mainly on aortic clamping and release, when bypass was initiated, and during defibrillation. Cognitive function deteriorated more in patients having cardiopulmonary bypass than in control patients having urologic operations but recovered in most tests by 2 months. Measures of cerebral perfusion (poor cerebrovascular reactivity, low arterial pressures, and flow velocity in the middle cerebral artery) predicted poor attention at 1 week (r = 0.3, P <.01, Spearman). Emboli were associated with memory loss (r = 0.3, P <.02, Spearman). CONCLUSIONS: Cognitive deficits were common after cardiopulmonary bypass. Occult cerebrovascular disease was more severe than expected and predisposed to attention difficulties, whereas emboli caused memory deficits. We believe this to be the first report of differing cognitive effects from emboli and hypoperfusion. PMID- 11385384 TI - Bidirectional cavopulmonary shunt with right ventricular outflow patency: the impact of pulsatility on pulmonary endothelial function. AB - OBJECTIVE: Although in vitro studies have suggested the importance of flow pulsatility in endothelial function, few reports have focused on pulmonary endothelial function under decreased pulsatile flow after a bidirectional cavopulmonary shunt with or without an additional pulmonary flow source. The purpose of the present study was to assess the pulmonary endothelial function after bidirectional cavopulmonary shunt. METHODS AND RESULTS: Pulmonary vasodilating response was evaluated in 10 patients 0.4 to 7.0 years (median 1.6 years) after bidirectional cavopulmonary shunt who were provided an additional flow source by retaining the pulmonary outflow tract and in 8 control subjects. Average pulmonary flow velocity was measured with a Doppler flow wire placed in the segmental lower lobe pulmonary artery during incremental infusion of acetylcholine (10(-8), 10(-7), 10(-6), and 10(-5) mol/L) and then of nitroglycerin (0.5 and 1.0 microg. kg(-1). min(-1)) after recovery. In the control subjects, a dose-dependent increase in flow velocity was observed in response to acetylcholine (maximum increase was 155% +/- 17% of baseline) and to nitroglycerin (maximum increase was 151% +/- 20% of baseline). In contrast, patients showed a significantly impaired response to acetylcholine (maximum increase was 124% +/- 17% of baseline; P <.01 vs control), whereas the response to nitroglycerin was preserved (138% +/- 12% of baseline; P =.09 vs control). In addition, the maximum response to acetylcholine correlated significantly with the pulmonary pulse pressure (r = 0.89, P <.01) and with the pulmonary flow pulsatility (r = 0.88, P <.01). CONCLUSIONS: These results clearly suggest that patients after bidirectional cavopulmonary shunt show pulmonary endothelial functional attenuation and, of more importance, that decreased pulsatility of cavopulmonary flow is mainly responsible for this endothelial abnormality. PMID- 11385385 TI - The push-and-pull mechanism to scavenge redox-active transition metals: a novel concept in myocardial protection. AB - OBJECTIVE: Traces of redox-active transition metals such as iron and copper play an important role in free radical formation during postischemic reperfusion of the heart. Two studies were conducted to assess the efficacy of the complexes of desferrioxamine with zinc or gallium to prevent this aspect of reperfusion injury. METHODS: In study I, isolated working rat hearts (n = 96) were subjected to 2 hours of hypothermic arrest at 10 degrees C induced by use of St Thomas' Hospital cardioplegic solution II supplemented with desferrioxamine, zinc histidinate, zinc-desferrioxamine, gallium-nitrate, or gallium-desferrioxamine. In study II, isolated nonworking rat hearts (n = 23) were subjected to normothermic regional (10 minutes) or global (35 minutes) unprotected ischemia. In this study, the perfusate was supplemented with gallium-desferrioxamine during preischemic and postischemic periods. RESULTS: In study I, the addition of desferrioxamine, zinc-histidinate, or gallium-nitrate to St Thomas' Hospital solution II improved postischemic aortic flow recovery. When the binary complexes zinc-desferrioxamine or gallium-desferrioxamine were added, however, functional recovery was further enhanced significantly. In study II, high-performance liquid chromatography analyses of tissue from postischemic hearts exposed to unsupplemented perfusate revealed a marked increase of malondialdehydes. In hearts perfused with perfusate supplemented with gallium-desferrioxamine, however, tissue malondialdehyde concentrations were significantly smaller, indicating reduced free radical formation. CONCLUSIONS: The data suggest synergistic protection by the complexes of the iron chelator desferrioxamine with zinc or gallium. The single components neutralize transition metals by 2 different but complementary push-and-pull mechanisms, thereby leading to an inhibition of metal-mediated site-specific free radical formation and improvement of postischemic cardiac function. PMID- 11385386 TI - Superior hepatic mitochondrial oxidation-reduction state in normothermic cardiopulmonary bypass. AB - OBJECTIVE: This study is the first comparative investigation of hepatic blood flow and oxygen metabolism during normothermic and hypothermic cardiopulmonary bypass. METHODS: Twenty-four patients undergoing coronary bypass operations were randomly divided into 2 groups according to their perfusion temperatures, either normothermia (36 degrees C) or hypothermia (30 degrees C). The clearance of indocyanine green was measured at 3 points. Arterial and hepatic venous ketone body ratios (an index of mitochondrial redox potential) and hepatic venous saturation were measured. RESULTS: Hepatic blood flow in both groups was identical before, during, and after cardiopulmonary bypass (normothermia, 499 +/- 111, 479 +/- 139, and 563 +/- 182 mL/min, respectively; hypothermia, 476 +/- 156, 491 +/- 147, and 560 +/- 202 mL/min, respectively). The hepatic venous saturation levels were significantly lower during cardiopulmonary bypass in the normothermic group (normothermia, 41% +/- 13%; hypothermia, 61% +/- 18%; P <.01), indicating a higher level of oxygen extraction use. The arterial ketone body ratio in the hypothermic group decreased severely after the onset of cardiopulmonary bypass (P <.01) and did not return to its subnormal value (>0.7) until the second postoperative day. However, the reduction in arterial ketone body ratio was less severe in the normothermic group. The difference in hepatic venous ketone body ratios was more obvious, and the hepatic venous ketone body ratios in the normothermic group were statistically superior to those of the hypothermic group throughout the course (P <.05-.01). CONCLUSIONS: Normothermic cardiopulmonary bypass provides adequate liver perfusion and results in a better hepatic mitochondrial redox potential than hypothermic cardiopulmonary bypass. Because arterial ketone body ratios reflect hepatic energy potential, normothermia was considered to be physiologically more advantageous for hepatic function. PMID- 11385387 TI - Transplantation of tracheal epithelial cells onto a prefabricated capsule pouch with fibrin glue as a delivery vehicle. AB - OBJECTIVE: The purpose of this study was to investigate whether in vitro cultured tracheal epithelial cells can be transplanted onto a prefabricated capsule surface in vivo for possible use in tracheal reconstruction. METHODS: Tracheal epithelial cells from 12 donor inbred rats were harvested for culture and expansion. In 16 recipient inbred rats, 2 sterile cylinders made of silicone rubber were implanted in each rat bilaterally in the folds of both the left and right anterior rectus sheath by wrapping the sheaths around the cylinders to induce a capsule formation. Ten days later, the cell cultures were divided and suspended in 1 of 2 delivery vehicles (standard culture medium or fibrin glue) and implanted onto the capsule surface. To compare the 2 delivery vehicles, we used fibrin glue on one side and the standard culture medium on the other. RESULTS: After 2 (group 1, n = 8) and 4 (group 2, n = 8) weeks, histologic findings, immunohistochemical staining, and electron microscopy demonstrated the capsule to be covered with a tracheal neoepithelium in group 1 and additional ciliated cells and secretory cells in a confluent layer in group 2 but only on the side with fibrin glue as the delivery vehicle. No viable epithelial cells were identified on the side with the standard culture medium in either group. CONCLUSION: We conclude that cultured epithelial cells can be successfully transplanted onto a prefabricated capsule surface with fibrin glue, which will differentiate into morphologic, nearly normal epithelium, showing potential for tracheal reconstruction. PMID- 11385388 TI - Bilobectomy for massive hemoptysis after bilateral lung transplantation. PMID- 11385390 TI - Automated implantable cardiac defibrillator and biventricular Thoratec assist device as bridge to transplantation in a patient with sarcoidosis. PMID- 11385389 TI - Endoscopic surgery with use of a laryngeal mask and a fiberoptic flexible bronchoscope. PMID- 11385391 TI - Hemodynamic evaluation for closing interatrial communication after fenestrated Fontan operation. PMID- 11385392 TI - Right ventricular outflow reconstruction with nonsutured pedicled autologous pericardium. PMID- 11385393 TI - Norwood operation for left isomeric heart with aortic atresia: evaluation with three-dimensional computed tomography. PMID- 11385394 TI - A new syndrome of multiple hemangiomas, right dominant double aortic arch, and coarctation. PMID- 11385395 TI - Surgical treatment of multiple aneurysms in a patient with Ehlers-Danlos syndrome. PMID- 11385396 TI - De novo inverted left atrial appendage: an unrecognized cause of left atrial mass with symptoms mimicking myxoma. PMID- 11385397 TI - Acquired aortopulmonary fistula in acute dissection. PMID- 11385398 TI - Acute retrograde aortic dissection during operations for ruptured Stanford type B dissection. PMID- 11385399 TI - Stent-graft entry closure and balloon fenestration for a case of aortic dissection accompanied by organ malperfusion. PMID- 11385400 TI - Sternal vascularity after harvesting of the internal thoracic artery. PMID- 11385402 TI - Valve-preserving aortic root reconstruction. PMID- 11385404 TI - Insulin cardioplegia for elective coronary bypass surgery. PMID- 11385406 TI - Extra-anatomic aortic bypass for thoracic aortic obstruction. PMID- 11385407 TI - Distal anastomotic site of extra-anatomic aortic bypass through a sternotomy for complex aortic arch obstruction. PMID- 11385411 TI - Overcoming the "yuk" factor. PMID- 11385409 TI - Results of surgical treatment of lung cancer involving the diaphragm. PMID- 11385413 TI - OECD says industrial biotech not realizing potential. PMID- 11385414 TI - Fragmentation is industry obstacle in Europe. PMID- 11385415 TI - UK keen to mimic German venture scheme. PMID- 11385417 TI - US biotech policy issues remain in limbo. PMID- 11385416 TI - Aurora throws light on Vertex's aspirations. PMID- 11385418 TI - US FTC probes practices of 100 companies. PMID- 11385433 TI - Complex biology with no parameters. PMID- 11385435 TI - Gene repair validation. PMID- 11385436 TI - Digital DNA signatures for animal tagging. PMID- 11385437 TI - PERV clarification. PMID- 11385442 TI - Biotechnology in the Baltic. PMID- 11385441 TI - Biotechnology Texas style. PMID- 11385443 TI - Italy, caught between old woes and new hopes. PMID- 11385444 TI - Israel's great expectations. PMID- 11385445 TI - Singapore--from microprocessors to microarrays? PMID- 11385446 TI - Ontario--hot biotechnology in a cold climate. PMID- 11385447 TI - Vaccines to die for. PMID- 11385448 TI - Selecting by microdialysis. PMID- 11385449 TI - Toward knockout sheep. PMID- 11385450 TI - Holistic approaches to glycobiology. PMID- 11385451 TI - Adding new meanings to the genetic code. PMID- 11385457 TI - Isolation of high-affinity ligand-binding proteins by periplasmic expression with cytometric screening (PECS). AB - Periplasmic expression with cytometric screening (PECS) is a powerful and rapid "display-less" technology for isolating ligand-binding proteins from diverse libraries. Escherichia coli expressing a library of proteins secreted into the periplasmic space are incubated with a fluorescent conjugate of the target ligand. Under the proper conditions, ligands as large as about 10 kDa can equilibrate within the periplasmic space without compromising the cell's integrity or viability. The bacterial cell envelope effectively serves as a dialysis bag to selectively retain receptor-fluorescent probe complexes but not free ligand. Cells displaying increased fluorescence are then isolated by flow cytometry. We demonstrate that scFv antibodies with both very high and low affinity to digoxigenin can be isolated from libraries screened by PECS using a benchtop flow cytometer. We also show that preexisting libraries constructed for display on filamentous bacteriophage can be screened by PECS without the need for subcloning. In fact, PECS was found to select for proteins that could be missed by conventional phage panning and screening methods. PMID- 11385458 TI - Apoptosis-mediated enhancement of DNA-raised immune responses by mutant caspases. AB - Apoptotic bodies can be used to target delivery of DNA-expressed immunogens into professional antigen-presenting cells (APCs). Here we show that antigen-laden apoptotic bodies created by vectors co-expressing influenza virus hemagglutinin (HA) or nucleoprotein (NP) genes and mutant caspase genes markedly increased T cell responses. Both CD8 and CD4 T-cell responses were affected. The adjuvant activity was restricted to partially inactivated caspases that allowed immunogen expression before the generation of apoptotic bodies. Active-site mutants of murine caspase 2 and an autocatalytic chimera of murine caspase 2 prodomain and human caspase 3 induced apoptosis that did not interfere with immunogen expression. The adjuvant activity also enhanced B-cell responses, but to a lesser extent than T-cell responses. The large increases in T-cell responses represent one of the strongest effects to date of a DNA adjuvant on cellular immunity. PMID- 11385459 TI - A plant-based multicomponent vaccine protects mice from enteric diseases. AB - Cholera toxin (CT) B and A2 subunit complementary DNAs (cDNAs) were fused to a rotavirus enterotoxin and enterotoxigenic Escherichia coli fimbrial antigen genes and transferred into potato. Immunoblot and enzyme-linked immunosorbent assay (ELISA) results indicated that the fusion antigens were synthesized in transformed tuber tissues and assembled into cholera holotoxin-like structures that retained enterocyte-binding affinity. Orally immunized mice generated detectable levels of serum and intestinal antibodies against the pathogen antigens. Elevated levels of interleukin 2 (IL2) and interferon gamma (INFgamma) detected in immunogen-challenged spleen cells from the immunized mice indicated the presence of a strong Th1 immune response to the three plant-synthesized antigens. This result was supported by flow cytometry analysis of immunized mouse spleen cells that showed a significant increase in CD4+ lymphocyte numbers. Diarrhea symptoms were reduced in severity and duration in passively immunized mouse neonates following rotavirus challenge. The results suggest that food plants can function as vaccines for simultaneous protection against infectious virus and bacterial diseases. PMID- 11385460 TI - Metabolic selection of glycosylation defects in human cells. AB - Changes in glycosylation are often associated with disease progression, but the genetic and metabolic basis of these events is rarely understood in detail at a molecular level. We describe a metabolism-based approach to the selection of mutants in glycoconjugate biosynthesis that provides insight into regulatory mechanisms for oligosaccharide expression and metabolic flux. Unnatural intermediates are used to challenge a specific pathway, and cell surface expression of their metabolic products provides a readout of flux in that pathway and a basis for selecting genetic mutants. The approach was applied to the sialic acid metabolic pathway in human cells, yielding novel mutants with phenotypes related to the inborn metabolic defect sialuria and metastatic tumor cells. PMID- 11385461 TI - Deletion of the alpha(1,3)galactosyl transferase (GGTA1) gene and the prion protein (PrP) gene in sheep. AB - Nuclear transfer offers a cell-based route for producing precise genetic modifications in a range of animal species. Using sheep, we report reproducible targeted gene deletion at two independent loci in fetal fibro-blasts. Vital regions were deleted from the alpha(1,3)galactosyl transferase (GGTA1) gene, which may account for the hyperacute rejection of xenografted organs, and from the prion protein (PrP) gene, which is directly associated with spongiform encephalopathies in humans and animals. Reconstructed embryos were prepared using cultures of targeted or nontargeted donor cells. Eight pregnancies were maintained to term and four PrP-/+ lambs were born. Although three of these perished soon after birth, one survived for 12 days. These data show that lambs carrying targeted gene deletions can be generated by nuclear transfer. PMID- 11385462 TI - In vitro abzyme evolution to optimize antibody recognition for catalysis. AB - Enzymes have evolved their ability to use binding energies for catalysis by increasing the affinity for the transition state of a reaction and decreasing the affinity for the ground state. To evolve abzymes toward higher catalytic activity, we have reconstructed an enzyme-evolutionary process in vitro. Thus, a phage-displayed combinatorial library from a hydrolytic abzyme, 6D9, generated by the conventional in vivo method with immunization of the transition-state analog (TSA), was screened against a newly devised TSA to optimize the differential affinity for the transition state relative to the ground state. The library format successfully afforded evolved variants with 6- to 20-fold increases in activity (kcat) as compared with 6D9. Structural analysis revealed an advantage of the in vitro evolution over the in vivo evolution: an induced catalytic residue in the evolved abzyme arises from double mutations in one codon, which rarely occur in somatic hypermutation in the immune response. PMID- 11385463 TI - A sensitive transgenic plant system to detect toxic inorganic compounds in the environment. AB - We describe a transgenic plant-based assay to study the genetic effects of heavy metals. Arabidopsis thaliana plants carrying a beta-glucuronidase (GUS) marker gene either with a point mutation or as a recombination substrate were used to analyze the frequency of somatic point mutations and homologous recombination in whole plants. Transgenic test plants sown on media contaminated by the salts of the heavy metals Cd2+, Pb2+, Ni2+, Zn2+, Cu2+, and As2O3 exhibited a pronounced uptake-dependent increase in the frequencies of both somatic intrachromosomal recombination and point mutation. The test was applied to monitor the genotoxicity of soils sampled in sites contaminated with several heavy metals. Our results indicate that this is a highly sensitive system for monitoring metal contamination in soils and water. PMID- 11385464 TI - Production of spider silk proteins in tobacco and potato. AB - Spider dragline silk is a proteinaceous fiber with remarkable mechanical properties that make it attractive for technical applications. Unfortunately, the material cannot be obtained in large quantities from spiders. We have therefore generated transgenic tobacco and potato plants that express remarkable amounts of recombinant Nephila clavipes dragline proteins. Using a gene synthesis approach, the recombinant proteins exhibit homologies of >90% compared to their native models. Here, we demonstrate the accumulation of recombinant silk proteins, which are encoded by synthetic genes of 420-3,600 base pairs, up to a level of at least 2% of total soluble protein in the endoplasmic reticulum (ER) of tobacco and potato leaves and potato tubers, respectively. Using the present expression system, spider silk proteins up to 100 kDa could be detected in plant tissues. When produced in plants, the recombinant spidroins exhibit extreme heat stability a property that is used to purify the spidroins by a simple and efficient procedure. PMID- 11385465 TI - A gene trap vector system for identifying transcriptionally responsive genes. AB - We present a method for fast and efficient trapping of genes whose transcription is regulated by exogenous stimuli. We constructed a promoterless retroviral vector transducing a green fluorescent protein-nitroreductase (GFNR) fusion protein downstream from a splice acceptor site. Flow cytometric analysis of the infected population allows identification and sorting of cells in which the trap is integrated downstream from an active promoter. Conversely, the nitroreductase (NTR) moiety allows pharmacological selection against constitutive GFNR expression. Using hepatocyte growth factor (HGF) stimulation of liver cells combined with either positive or negative selection, we recovered cell populations carrying traps in induced or suppressed genes, respectively. Several distinct responsive clones were isolated, and regulated expression of the trapped gene was confirmed at the RNA level. Positive and negative selection can be calibrated to recover traps in genes showing different levels of basal expression or transcriptional regulation. The flexibility and efficiency of the GFNR-based trap screening procedure make it suitable for wide surveys of transcriptionally regulated genes. PMID- 11385466 TI - Efficient FLPe recombinase enables scalable production of helper-dependent adenoviral vectors with negligible helper-virus contamination. AB - Helper-dependent (HD), high-capacity adenoviruses are one of the most efficient and safe gene therapy vectors, capable of mediating long-term expression. Currently, the most widely used system for HD vector production avoids significant contamination with helper virus by using producer cells stably expressing a nuclear-targeted Cre recombinase and an engineered first-generation helper virus with parallel loxP sites flanking its packaging signal. The system requires a final, density-based separation of HD and residual helper viruses by ultracentrifugation to reduce contaminating helper virus to low levels. This separation step hinders large-scale production of clinical-grade HD virus. By using a very efficient recombinase, in vitro-evolved FLPe (ref. 14), to excise the helper virus packaging signal in the producer cells, we have developed a scalable HD vector production method. FLP has previously been shown to mediate maximum levels of excision close to 100% compared to 80% for Cre (ref. 15). Utilizing a common HD plasmid backbone, the FLPe-based system reproducibly yielded HD virus with the same low levels of helper virus contamination before any density-based separation by ultracentrifugation. This should allow large scale production of HD vectors using column chromatography-based virus purification. PMID- 11385467 TI - The Monsanto decision: the edge or the wedge. PMID- 11385471 TI - Hiring foreign workers: what you need to know. PMID- 11385472 TI - Sex matters. PMID- 11385473 TI - Quick success for cancer kinase treatment. PMID- 11385474 TI - Defective DNA polymerase-delta proofreading causes cancer susceptibility in mice. PMID- 11385475 TI - Still more debate over VEGF. PMID- 11385477 TI - Is EDRF a specific marker for TSEs? PMID- 11385479 TI - Dark clouds over Toronto psychiatry research. PMID- 11385480 TI - Olivieri to testify against Apotex in Europe. PMID- 11385481 TI - Proposed Bush budget favors NIH. PMID- 11385482 TI - Scientists question rise in autism. PMID- 11385483 TI - Europe to standardize bacteria research. PMID- 11385484 TI - Web bounty hunters chase research patents [correction of patients]. PMID- 11385485 TI - FDA tightens human oversight. PMID- 11385487 TI - May brings money for Malaria research. PMID- 11385488 TI - Annan gives prize money to AIDS Fund. PMID- 11385489 TI - Financial houses scared off by vivisectionist threats. PMID- 11385491 TI - Rod MacKinnon. PMID- 11385492 TI - The allogeneic response and tumor immunity. AB - The strong allogeneic response to donor MHC molecules in transplantation and the weak response to tumor antigens represent two important and divergent but potentially interactive immune responses. A patient's response to allogeneic MHC molecules might promote an effective T-cell response to self MHC-restricted tumor peptides and the possibilities for this are discussed here. These allogeneic responses might successfully be harnessed to promote the immune eradication of metastatic cancer. PMID- 11385494 TI - Imaging metalloproteinase activity in vivo. PMID- 11385495 TI - A protective player in the vascular response to injury. PMID- 11385496 TI - Microarrays--the 21st century divining rod? PMID- 11385497 TI - A dominant-negative therapy for anthrax. PMID- 11385498 TI - Can the thymus win the battle against drug-resistant HIV? PMID- 11385499 TI - Hot on the TRAIL of acute promyelocytic leukemia. PMID- 11385500 TI - Clearing the way to mechanisms of autoimmunity. PMID- 11385502 TI - Advances in schizophrenia. AB - Recent studies into the etiology of schizophrenia have yielded both promising leads and disappointing dead ends, indicating the multifactored and complex nature of the disorder. The focus has subsequently shifted back to refining the phenotype and identifying clinical and biological subtypes. Recent technological breakthroughs in genomics and proteomics hold promise for advancing our understanding of the molecular pathophysiology of schizophrenia. PMID- 11385503 TI - Classification and diagnostic prediction of cancers using gene expression profiling and artificial neural networks. AB - The purpose of this study was to develop a method of classifying cancers to specific diagnostic categories based on their gene expression signatures using artificial neural networks (ANNs). We trained the ANNs using the small, round blue-cell tumors (SRBCTs) as a model. These cancers belong to four distinct diagnostic categories and often present diagnostic dilemmas in clinical practice. The ANNs correctly classified all samples and identified the genes most relevant to the classification. Expression of several of these genes has been reported in SRBCTs, but most have not been associated with these cancers. To test the ability of the trained ANN models to recognize SRBCTs, we analyzed additional blinded samples that were not previously used for the training procedure, and correctly classified them in all cases. This study demonstrates the potential applications of these methods for tumor diagnosis and the identification of candidate targets for therapy. PMID- 11385504 TI - Retinoic acid-induced apoptosis in leukemia cells is mediated by paracrine action of tumor-selective death ligand TRAIL. AB - The therapeutic and preventive activities of retinoids in cancer are due to their ability to modulate the growth, differentiation, and survival or apoptosis of cancer cells. Here we show that in NB4 acute promyelocytic leukemia cells, retinoids selective for retinoic-acid receptor-alpha induced an autoregulatory circuitry of survival programs followed by expression of the membrane-bound tumor selective death ligand, TRAIL (tumor necrosis factor-related apoptosis-inducing ligand, also called Apo-2L). In a paracrine mode of action, TRAIL killed NB4 as well as heterologous and retinoic-acid-resistant cells. In the leukemic blasts of freshly diagnosed acute promyelocytic leukemia patients, retinoic-acid-induced expression of TRAIL most likely caused blast apoptosis. Thus, induction of TRAIL mediated death signaling appears to contribute to the therapeutic value of retinoids. PMID- 11385505 TI - Statins selectively inhibit leukocyte function antigen-1 by binding to a novel regulatory integrin site. AB - The beta2 integrin leukocyte function antigen-1 (LFA-1) has an important role in the pathophysiology of inflammatory and autoimmune diseases. Here we report that statin compounds commonly used for the treatment of hypercholesterolemia selectively blocked LFA-1-mediated adhesion and costimulation of lymphocytes. This effect was unrelated to the statins' inhibition of 3-hydroxy-3 methylglutaryl coenzyme-A reductase; instead it occurred via binding to a novel allosteric site within LFA-1. Subsequent optimization of the statins for LFA-1 binding resulted in potent, selective and orally active LFA-1 inhibitors that suppress the inflammatory response in a murine model of peritonitis. Targeting of the statin-binding site of LFA-1 could be used to treat diseases such as psoriasis, rheumatoid arthritis, ischemia/reperfusion injury and transplant rejection. PMID- 11385506 TI - Heme oxygenase-1 protects against vascular constriction and proliferation. AB - Heme oxygenase (HO-1, encoded by Hmox1) is an inducible protein activated in systemic inflammatory conditions by oxidant stress. Vascular injury is characterized by a local reparative process with inflammatory components, indicating a potential protective role for HO-1 in arterial wound repair. Here we report that HO-1 directly reduces vasoconstriction and inhibits cell proliferation during vascular injury. Expression of HO-1 in arteries stimulated vascular relaxation, mediated by guanylate cyclase and cGMP, independent of nitric oxide. The unexpected effects of HO-1 on vascular smooth muscle cell growth were mediated by cell-cycle arrest involving p21Cip1. HO-1 reduced the proliferative response to vascular injury in vivo; expression of HO-1 in pig arteries inhibited lesion formation and Hmox1-/- mice produced hyperplastic arteries compared with controls. Induction of the HO-1 pathway moderates the severity of vascular injury by at least two adaptive mechanisms independent of nitric oxide, and is a potential therapeutic target for diseases of the vasculature. PMID- 11385508 TI - The morphogen Sonic hedgehog is an indirect angiogenic agent upregulating two families of angiogenic growth factors. AB - Sonic hedgehog (Shh) is a prototypical morphogen known to regulate epithelial/mesenchymal interactions during embryonic development. We found that the hedgehog-signaling pathway is present in adult cardiovascular tissues and can be activated in vivo. Shh was able to induce robust angiogenesis, characterized by distinct large-diameter vessels. Shh also augmented blood-flow recovery and limb salvage following operatively induced hind-limb ischemia in aged mice. In vitro, Shh had no effect on endothelial-cell migration or proliferation; instead, it induced expression of two families of angiogenic cytokines, including all three vascular endothelial growth factor-1 isoforms and angiopoietins-1 and -2 from interstitial mesenchymal cells. These findings reveal a novel role for Shh as an indirect angiogenic factor regulating expression of multiple angiogenic cytokines and indicate that Shh might have potential therapeutic use for ischemic disorders. PMID- 11385507 TI - Lack of macrophage fatty-acid-binding protein aP2 protects mice deficient in apolipoprotein E against atherosclerosis. AB - The adipocyte fatty-acid-binding protein, aP2, has an important role in regulating systemic insulin resistance and lipid metabolism. Here we demonstrate that aP2 is also expressed in macrophages, has a significant role in their biological responses and contributes to the development of atherosclerosis. Apolipoprotein E (ApoE)-deficient mice also deficient for aP2 showed protection from atherosclerosis in the absence of significant differences in serum lipids or insulin sensitivity. aP2-deficient macrophages showed alterations in inflammatory cytokine production and a reduced ability to accumulate cholesterol esters when exposed to modified lipoproteins. Apoe-/- mice with Ap2+/+ adipocytes and Ap2-/- macrophages generated by bone-marrow transplantation showed a comparable reduction in atherosclerotic lesions to those with total aP2 deficiency, indicating an independent role for macrophage aP2 in atherogenesis. Through its distinct actions in adipocytes and macrophages, aP2 provides a link between features of the metabolic syndrome and could be a new therapeutic target for the prevention of atherosclerosis. PMID- 11385509 TI - Impaired replication of protease inhibitor-resistant HIV-1 in human thymus. AB - Many HIV-1-infected patients treated with protease inhibitors (PI) develop PI resistant HIV-1 variants and rebounds in viremia, but their CD4+ T-cell counts often do not fall. We hypothesized that in these patients, T-cell counts remain elevated because PI-resistant virus spares intrathymic T-cell production. To test this, we studied recombinant HIV-1 clones containing wild-type or PI-resistant protease domains, as well as uncloned isolates from patients, in activated peripheral blood mononuclear cells, human thymic organ cultures and human thymus implants in SCID-hu Thy/Liv mice. In most cases, wild-type and PI-resistant HIV-1 isolates replicated to similar degrees in peripheral blood mononuclear cells. However, the replication of PI-resistant but not wild-type HIV-1 isolates was highly impaired in thymocytes. In addition, patients who had PI-resistant HIV-1 had abundant thymus tissue as assessed by computed tomography. We propose that the inability of PI-resistant HIV-1 to replicate efficiently in thymus contributes to the preservation of CD4+ T-cell counts in patients showing virologic rebound on PI therapy. PMID- 11385510 TI - A newly discovered human pneumovirus isolated from young children with respiratory tract disease. AB - From 28 young children in the Netherlands, we isolated a paramyxovirus that was identified as a tentative new member of the Metapneumovirus genus based on virological data, sequence homology and gene constellation. Previously, avian pneumovirus was the sole member of this recently assigned genus, hence the provisional name for the newly discovered virus: human metapneumovirus. The clinical symptoms of the children from whom the virus was isolated were similar to those caused by human respiratory syncytial virus infection, ranging from upper respiratory tract disease to severe bronchiolitis and pneumonia. Serological studies showed that by the age of five years, virtually all children in the Netherlands have been exposed to human metapneumovirus and that the virus has been circulating in humans for at least 50 years. PMID- 11385511 TI - Disruption of Akt kinase activation is important for immunosuppression induced by measles virus. AB - Surface-contact-mediated signaling induced by the measles virus (MV) fusion and hemagglutinin glycoproteins is necessary and sufficient to induce T-cell unresponsiveness in vitro and in vivo. To define the intracellular pathways involved, we analyzed interleukin (IL)-2R signaling in primary human T cells and in Kit-225 cells. Unlike IL-2-dependent activation of JAK/STAT pathways, activation of Akt kinase was impaired after MV contact both in vitro and in vivo. MV interference with Akt activation was important for immunosuppression, as expression of a catalytically active Akt prevented negative signaling by the MV glycoproteins. Thus, we show here that MV exploits a novel strategy to interfere with T-cell activation during immunosuppression. PMID- 11385512 TI - Overexpression of heat-shock proteins reduces survival of Mycobacterium tuberculosis in the chronic phase of infection. AB - Elevated expression of heat-shock proteins (HSPs) can benefit a microbial pathogen struggling to penetrate host defenses during infection, but at the same time might provide a crucial signal alerting the host immune system to its presence. To determine which of these effects predominate, we constructed a mutant strain of Mycobacterium tuberculosis that constitutively overexpresses Hsp70 proteins. Although the mutant was fully virulent in the initial stage of infection, it was significantly impaired in its ability to persist during the subsequent chronic phase. Induction of microbial genes encoding HSPs might provide a novel strategy to boost the immune response of individuals with latent tuberculosis infection. PMID- 11385513 TI - Host bone-marrow cells are a source of donor intimal smooth- muscle-like cells in murine aortic transplant arteriopathy. AB - Long-term solid-organ allografts typically develop diffuse arterial intimal lesions (graft arterial disease; GAD), consisting of smooth-muscle cells (SMC), extracellular matrix and admixed mononuclear leukocytes. GAD eventually culminates in vascular stenosis and ischemic graft failure. Although the exact mechanisms are unknown, chronic low-level alloresponses likely induce inflammatory cells and/or dysfunctional vascular wall cells to secrete growth factors that promote SMC intimal recruitment, proliferation and matrix synthesis. Although prior work demonstrated that the endothelium and medial SMCs lining GAD lesions in cardiac allografts are donor-derived, the intimal SMC origin could not be determined. They are generally presumed to originate from the donor media, leading to interventions that target donor medial SMC proliferation, with limited efficacy. However, other reports indicate that allograft vessels may contain host derived endothelium and SMCs (refs. 8,9). Moreover, subpopulations of bone-marrow and circulating cells can differentiate into endothelium, and implanted synthetic vascular grafts are seeded by host SMCs and endothelium. Here we used murine aortic transplants to formally identify the source of SMCs in GAD lesions. Allografts in beta-galactosidase transgenic recipients showed that intimal SMCs derived almost exclusively from host cells. Bone-marrow transplantation of beta galactosidase--expressing cells into aortic allograft recipients demonstrated that intimal cells included those of marrow origin. Thus, smooth-muscle--like cells in GAD lesions can originate from circulating bone--marrow-derived precursors. PMID- 11385516 TI - Retraction: SADS: A new component of Fas-DISC is the accelerator for cell death signaling and is downregulated in patients with colon carcinoma. PMID- 11385514 TI - In vivo molecular target assessment of matrix metalloproteinase inhibition. AB - A number of different matrix metalloproteinase (MMP) inhibitors have been developed as cytostatic and anti-angiogenic agents and are currently in clinical testing. One major hurdle in assessing the efficacy of such drugs has been the inability to sense or image anti-proteinase activity directly and non-invasively in vivo. We show here that novel, biocompatible near-infrared fluorogenic MMP substrates can be used as activatable reporter probes to sense MMP activity in intact tumors in nude mice. Moreover, we show for the first time that the effect of MMP inhibition can be directly imaged using this approach within hours after initiation of treatment using the potent MMP inhibitor, prinomastat (AG3340). The developed probes, together with novel near-infrared fluorescence imaging technology will enable the detailed analysis of a number of proteinases critical for advancing the therapeutic use of clinical proteinase inhibitors. PMID- 11385519 TI - Europe's right approach to energy. PMID- 11385518 TI - Dangers of nationalism. PMID- 11385521 TI - Arson hampers conservation work. PMID- 11385520 TI - Data protection law threatens to derail UK epidemiology studies. PMID- 11385522 TI - Senate shift bodes well for 'green' science. PMID- 11385523 TI - 'Contrary' trade sanctions worry malaria researchers. PMID- 11385526 TI - Eastern Europe decries EU research proposal. PMID- 11385524 TI - NIH faces double trouble over budget rise. PMID- 11385528 TI - Gene tests lift lid on drug-resistance puzzle. PMID- 11385529 TI - Bioinformatics to be nurtured online. PMID- 11385531 TI - A change of climate for big oil. PMID- 11385532 TI - The North Sea bubble. PMID- 11385533 TI - Toronto's science jewel. PMID- 11385534 TI - Free online availability substantially increases a paper's impact. PMID- 11385535 TI - Authors willing to pay for instant web access. PMID- 11385536 TI - Impact factors, and why they won't go away. PMID- 11385538 TI - The Net is many people's only chance of access. PMID- 11385537 TI - Evolution is what's needed, not revolution. PMID- 11385544 TI - Formally speaking. PMID- 11385545 TI - Knife-edge of design. PMID- 11385546 TI - Entrepreneurial insects. PMID- 11385548 TI - Superconductivity. Rehearsals for prime time. PMID- 11385550 TI - Earth science. Mantle cookbook calibration. PMID- 11385549 TI - Developmental biology. Fishing for morphogens. PMID- 11385551 TI - Behavioural ecology. Down on fungal farm. PMID- 11385553 TI - Chemistry. A new twist on molecular shape. PMID- 11385552 TI - Human genetics. Tackling common disease. PMID- 11385554 TI - Immunology. Brief encounter. PMID- 11385555 TI - Cell biology. Channels as enzymes. PMID- 11385557 TI - Obituary. G.N. Ramachandran (1922-2001). PMID- 11385558 TI - Natural selection and resistance to HIV. PMID- 11385559 TI - Climate change. Increasing shrub abundance in the Arctic. AB - The warming of the Alaskan Arctic during the past 150 years has accelerated over the last three decades and is expected to increase vegetation productivity in tundra if shrubs become more abundant; indeed, this transition may already be under way according to local plot studies and remote sensing. Here we present evidence for a widespread increase in shrub abundance over more than 320 km of Arctic landscape during the past 50 years, based on a comparison of historic and modern aerial photographs. This expansion will alter the partitioning of energy in summer and the trapping and distribution of snow in winter, as well as increasing the amount of carbon stored in a region that is believed to be a net source of carbon dioxide. PMID- 11385560 TI - Sensory adaptation. Tunable colour vision in a mantis shrimp. AB - Systems of colour vision are normally identical in all members of a species, but a single design may not be adequate for species living in a diverse range of light environments. Here we show that in the mantis shrimp Haptosquilla trispinosa, which occupies a range of depths in the ocean, long-wavelength colour receptors are individually tuned to the local light environment. The spectral sensitivity of specific classes of photoreceptor is adjusted by filters that vary between individuals. PMID- 11385561 TI - Developmental biology. Lungfish dental pattern conserved for 360 Myr. AB - Lungfish, the closest living relatives of four-limbed animals, are unique in that adults lack marginal teeth and have to rely on palatal dental plates for crushing food. We have discovered that an identical pattern of tooth development is used to shape these plates in the hatchlings of fossil and living lungfish species that are separated by 360 million years (Myr) of evolution, even though the adults have very different dental forms; the same pattern is also evident in the transient marginal dentition, despite being functional only until the juvenile stage. This remarkable finding indicates that developmental programming for dentition in lungfish is uniform, unique and conserved for all tooth fields. PMID- 11385562 TI - Many-body and correlation effects in semiconductors. AB - Solids consist of 1022-1023 particles per cubic centimetre, interacting through infinite-range Coulomb interactions. The linear response of a solid to a weak external perturbation is well described by the concept of non-interacting 'quasiparticles' first introduced by Landau. But interactions between quasiparticles can be substantial in dense systems. For example, studies over the past decade have shown that Coulomb correlations between quasiparticles dominate the nonlinear optical response of semiconductors, in marked contrast to the behaviour of atomic systems. These Coulomb correlations and other many-body interactions are important not only for semiconductors, but also for all condensed-matter systems. PMID- 11385563 TI - High critical current density and enhanced irreversibility field in superconducting MgB2 thin films. AB - The discovery of superconductivity at 39 K in magnesium diboride offers the possibility of a new class of low-cost, high-performance superconducting materials for magnets and electronic applications. This compound has twice the transition temperature of Nb3Sn and four times that of Nb-Ti alloy, and the vital prerequisite of strongly linked current flow has already been demonstrated. One possible drawback, however, is that the magnetic field at which superconductivity is destroyed is modest. Furthermore, the field which limits the range of practical applications-the irreversibility field H*(T)-is approximately 7 T at liquid helium temperature (4.2 K), significantly lower than about 10 T for Nb-Ti (ref. 6) and approximately 20 T for Nb3Sn (ref. 7). Here we show that MgB2 thin films that are alloyed with oxygen can exhibit a much steeper temperature dependence of H*(T) than is observed in bulk materials, yielding an H* value at 4.2 K greater than 14 T. In addition, very high critical current densities at 4.2 K are achieved: 1 MA cm-2 at 1 T and 105 A cm-2 at 10 T. These results demonstrate that MgB2 has potential for high-field superconducting applications. PMID- 11385564 TI - Enhancement of the high-magnetic-field critical current density of superconducting MgB2 by proton irradiation. AB - Magnesium diboride, MgB2, has a relatively high superconducting transition temperature, placing it between the families of low- and high-temperature (copper oxide based) superconductors. Supercurrent flow in MgB2 is unhindered by grain boundaries, making it potentially attractive for technological applications in the temperature range 20-30 K. But in the bulk material, the critical current density (Jc) drops rapidly with increasing magnetic field strength. The magnitude and field dependence of the critical current are related to the presence of structural defects that can 'pin' the quantized magnetic vortices that permeate the material, and a lack of natural defects in MgB2 may be responsible for the rapid decline of Jc with increasing field strength. Here we show that modest levels of atomic disorder induced by proton irradiation enhance the pinning of vortices, thereby significantly increasing Jc at high field strengths. We anticipate that either chemical doping or mechanical processing should generate similar levels of disorder, and so achieve performance that is technologically attractive in an economically viable way. PMID- 11385565 TI - High critical currents in iron-clad superconducting MgB2 wires. AB - Technically useful bulk superconductors must have high transport critical current densities, Jc, at operating temperatures. They also require a normal metal cladding to provide parallel electrical conduction, thermal stabilization, and mechanical protection of the generally brittle superconductor cores. The recent discovery of superconductivity at 39 K in magnesium diboride (MgB2) presents a new possibility for significant bulk applications, but many critical issues relevant for practical wires remain unresolved. In particular, MgB2 is mechanically hard and brittle and therefore not amenable to drawing into the desired fine-wire geometry. Even the synthesis of moderately dense, bulk MgB2 attaining 39 K superconductivity is a challenge because of the volatility and reactivity of magnesium. Here we report the successful fabrication of dense, metal-clad superconducting MgB2 wires, and demonstrate a transport Jc in excess of 85,000 A cm-2 at 4.2 K. Our iron-clad fabrication technique takes place at ambient pressure, yet produces dense MgB2 with little loss of stoichiometry. While searching for a suitable cladding material, we found that other materials dramatically reduced the critical current, showing that although MgB2 itself does not show the 'weak-link' effect characteristic of the high-Tc superconductors, contamination does result in weak-link-like behaviour. PMID- 11385566 TI - Hyperconjugation not steric repulsion leads to the staggered structure of ethane. AB - Many molecules can rotate internally around one or more of their bonds so that during a full 360 degrees rotation, they will change between unstable and relatively stable conformations. Ethane is the textbook example of a molecule exhibiting such behaviour: as one of its two methyl (CH3) groups rotates once around the central carbon-carbon bond, the molecule will alternate three times between an unstable eclipsed conformation and the preferred staggered conformation. This structural preference is usually attributed to steric effects; that is, while ethane rotates towards an eclipsed structure, the electrons in C-H bonds on the different C atoms are drawing closer to each other and therefore experience increased repulsion, introducing a rotation barrier that destabilizes the eclipsed structure. Stabilization of the staggered structure through rotation induced weakening of the central C-C bond and hyperconjugation has been considered to be involved, but evaluation of the contributions of these effects to ethane's internal rotation barrier and conformational preference remains difficult. Here we report a series of ethane structure optimizations, where successive removal of different interactions indicates that ethane's staggered conformation is the result of preferential stabilization through hyperconjugation. Removal of hyperconjugation interactions yields the eclipsed structure as the preferred conformation, whereas repulsive forces, either present or absent, have no influence on the preference for a staggered conformation. PMID- 11385567 TI - Possible displacement of the climate signal in ancient ice by premelting and anomalous diffusion. AB - The best high-resolution records of climate over the past few hundred millennia are derived from ice cores retrieved from Greenland and Antarctica. The interpretation of these records relies on the assumption that the trace constituents used as proxies for past climate have undergone only modest post depositional migration. Many of the constituents are soluble impurities found principally in unfrozen liquid that separates the grain boundaries in ice sheets. This phase behaviour, termed premelting, is characteristic of polycrystalline material. Here we show that premelting influences compositional diffusion in a manner that causes the advection of impurity anomalies towards warmer regions while maintaining their spatial integrity. Notwithstanding chemical reactions that might fix certain species against this prevailing transport, we find that under conditions that resemble those encountered in the Eemian interglacial ice of central Greenland (from about 125,000 to 115,000 years ago)-impurity fluctuations may be separated from ice of the same age by as much as 50 cm. This distance is comparable to the ice thickness of the contested sudden cooling events in Eemian ice from the GRIP core. PMID- 11385568 TI - The post-spinel transformation in Mg2SiO4 and its relation to the 660-km seismic discontinuity. AB - The 660-km seismic discontinuity in the Earth's mantle has long been identified with the transformation of (Mg,Fe)2SiO4 from gamma-spinel (ringwoodite) to (Mg,Fe)SiO3-perovskite and (Mg,Fe)O-magnesiowustite. This has been based on experimental studies of materials quenched from high pressure and temperature, which have shown that the transformation is consistent with the seismically observed sharpness and the depth of the discontinuity at expected mantle temperatures. But the first in situ examination of this phase transformation in Mg2SiO4 using a multi-anvil press indicated that the transformation occurs at a pressure about 2 GPa lower than previously thought (equivalent to approximately 600 km depth) and hence that it may not be associated with the 660-km discontinuity. Here we report the results of an in situ study of Mg2SiO4 at pressures of 20-36 GPa using a combination of double-sided laser-heating and synchrotron X-ray diffraction in a diamond-anvil cell. The phase transformation from gamma-Mg2SiO4 to MgSiO3-perovskite and MgO (periclase) is readily observed in both the forward and reverse directions. In contrast to the in situ multi anvil-press study, we find that the pressure and temperature of the post-spinel transformation in Mg2SiO4 is consistent with seismic observations for the 660-km discontinuity. PMID- 11385569 TI - High-pressure polymorphs of olivine and the 660-km seismic discontinuity. AB - It had long been accepted that the 400-km seismic discontinuity in the Earth's mantle results from the phase transition of (Mg,Fe)2-SiO4-olivine to its high pressure polymorph beta-spinel (wadsleyite), and that the 660-km discontinuity results from the breakdown of the higher-pressure polymorph gamma-spinel (ringwoodite) to MgSiO3-perovskite and (Mg,Fe)O-magnesiowustite. An in situ multi anvil-press X-ray study indicated, however, that the phase boundary of the latter transition occurs at pressures 2 GPa lower than had been found in earlier studies using multi-anvil recovery experiments and laser-heated diamond-anvil cells. Such a lower-pressure phase boundary would be irreconcilable with the accuracy of seismic measurements of the 660-km discontinuity, and would thus require a mineral composition of the mantle that is significantly different from what is currently thought. Here, however, we present measurements made with a laser heated diamond-anvil cell which indicate that gamma-Mg2SiO4 is stable up to pressure and temperature conditions equivalent to 660-km depth in the Earth's mantle (24 GPa and 1,900 K) and then breaks down into MgSiO3-perovskite and MgO (periclase). We paid special attention to pressure accuracy and thermal pressure in our experiments, and to ensuring that our experiments were performed under nearly hydrostatic, inert pressure conditions using a variety of heating methods. We infer that these factors are responsible for the different results obtained in our experiments compared to the in situ multi-anvil-press study. PMID- 11385570 TI - Ecological and evolutionary processes at expanding range margins. AB - Many animals are regarded as relatively sedentary and specialized in marginal parts of their geographical distributions. They are expected to be slow at colonizing new habitats. Despite this, the cool margins of many species' distributions have expanded rapidly in association with recent climate warming. We examined four insect species that have expanded their geographical ranges in Britain over the past 20 years. Here we report that two butterfly species have increased the variety of habitat types that they can colonize, and that two bush cricket species show increased fractions of longer-winged (dispersive) individuals in recently founded populations. Both ecological and evolutionary processes are probably responsible for these changes. Increased habitat breadth and dispersal tendencies have resulted in about 3- to 15-fold increases in expansion rates, allowing these insects to cross habitat disjunctions that would have represented major or complete barriers to dispersal before the expansions started. The emergence of dispersive phenotypes will increase the speed at which species invade new environments, and probably underlies the responses of many species to both past and future climate change. PMID- 11385571 TI - Honeybee dances communicate distances measured by optic flow. AB - In honeybees, employed foragers recruit unemployed hive mates to food sources by dances from which a human observer can read the distance and direction of the food source. When foragers collect food in a short, narrow tunnel, they dance as if the food source were much farther away. Dancers gauge distance by retinal image flow on the way to their destination. Their visually driven odometer misreads distance because the close tunnel walls increase optic flow. We examined how hive mates interpret these dances. Here we show that recruited bees search outside in the direction of the tunnel at exaggerated distances and not inside the tunnel where the foragers come from. Thus, dances must convey information about the direction of the food source and the total amount of image motion en route to the food source, but they do not convey information about absolute distances. We also found that perceived distances on various outdoor routes from the same hive could be considerably different. Navigational errors are avoided as recruits and dancers tend to fly in the same direction. Reported racial differences in honeybee dances could have arisen merely from differences in the environments in which these bees flew. PMID- 11385572 TI - Single cocaine exposure in vivo induces long-term potentiation in dopamine neurons. AB - How do drugs of abuse modify neural circuitry and thereby lead to addictive behaviour? As for many forms of experience-dependent plasticity, modifications in glutamatergic synaptic transmission have been suggested to be particularly important. Evidence of such changes in response to in vivo administration of drugs of abuse is lacking, however. Here we show that a single in vivo exposure to cocaine induces long-term potentiation of AMPA (alpha-amino-3-hydroxy-5-methyl isoxazole propionic acid)-receptor-mediated currents at excitatory synapses onto dopamine cells in the ventral tegmental area. Potentiation is still observed 5 but not 10 days after cocaine exposure and is blocked when an NMDA (N-methyl-d aspartate) receptor antagonist is administered with cocaine. Furthermore, long term potentiation at these synapses is occluded and long-term depression is enhanced by in vivo cocaine exposure. These results show that a prominent form of synaptic plasticity can be elicited by a single in vivo exposure to cocaine and therefore may be involved in the early stages of the development of drug addiction. PMID- 11385573 TI - Presynaptic glycine receptors enhance transmitter release at a mammalian central synapse. AB - Glycine and GABAA (gamma-aminobutyric acid A) receptors are inhibitory neurotransmitter-gated Cl- channels localized in postsynaptic membranes. In some cases, GABAA receptors are also found presynaptically, but they retain their inhibitory effect as their activation reduces excitatory transmitter release. Here we report evidence for presynaptic ionotropic glycine receptors, using pre- and postsynaptic recordings of a calyceal synapse in the medial nucleus of the trapezoid body (MNTB). Unlike the classical action of glycine, presynaptic glycine receptors triggered a weakly depolarizing Cl- current in the nerve terminal. The depolarization enhanced transmitter release by activating Ca2+ channels and increasing resting intraterminal Ca2+ concentrations. Repetitive activation of glycinergic synapses on MNTB neurons also enhanced glutamatergic synaptic currents, indicating that presynaptic glycine receptors are activated by glycine spillover. These results reveal a novel site of action of the transmitter glycine, and indicate that under certain conditions presynaptic Cl- channels may increase transmitter release. PMID- 11385574 TI - LTRPC7 is a Mg.ATP-regulated divalent cation channel required for cell viability. AB - The molecular mechanisms that regulate basal or background entry of divalent cations into mammalian cells are poorly understood. Here we describe the cloning and functional characterization of a Ca2+- and Mg2+-permeable divalent cation channel, LTRPC7 (nomenclature compatible with that proposed in ref. 1), a new member of the LTRPC family of putative ion channels. Targeted deletion of LTRPC7 in DT-40 B cells was lethal, indicating that LTRPC7 has a fundamental and nonredundant role in cellular physiology. Electrophysiological analysis of HEK 293 cells overexpressing recombinant LTRPC7 showed large currents regulated by millimolar levels of intracellular Mg.ATP and Mg.GTP with the permeation properties of a voltage-independent divalent cation influx pathway. Analysis of several cultured cell types demonstrated small magnesium-nucleotide-regulated metal ion currents (MagNuM) with regulation and permeation properties essentially identical to the large currents observed in cells expressing recombinant LTRPC7. Our data indicate that LTRPC7, by virtue of its sensitivity to physiological Mg.ATP levels, may be involved in a fundamental process that adjusts plasma membrane divalent cation fluxes according to the metabolic state of the cell. PMID- 11385575 TI - ADP-ribose gating of the calcium-permeable LTRPC2 channel revealed by Nudix motif homology. AB - Free ADP-ribose (ADPR), a product of NAD hydrolysis and a breakdown product of the calcium-release second messenger cyclic ADPR (cADPR), has no defined role as an intracellular signalling molecule in vertebrate systems. Here we show that a 350-amino-acid protein (designated NUDT9) and a homologous domain (NUDT9 homology domain) near the carboxy terminus of the LTRPC2/TrpC7 putative cation channel both function as specific ADPR pyrophosphatases. Whole-cell and single-channel analysis of HEK-293 cells expressing LTRPC2 show that LTRPC2 functions as a calcium-permeable cation channel that is specifically gated by free ADPR. The expression of native LTRPC2 transcripts is detectable in many tissues including the U937 monocyte cell line, in which ADPR induces large cation currents (designated IADPR) that closely match those mediated by recombinant LTRPC2. These results indicate that intracellular ADPR regulates calcium entry into cells that express LTRPC2. PMID- 11385576 TI - Association of NOD2 leucine-rich repeat variants with susceptibility to Crohn's disease. AB - Crohn's disease and ulcerative colitis, the two main types of chronic inflammatory bowel disease, are multifactorial conditions of unknown aetiology. A susceptibility locus for Crohn's disease has been mapped to chromosome 16. Here we have used a positional-cloning strategy, based on linkage analysis followed by linkage disequilibrium mapping, to identify three independent associations for Crohn's disease: a frameshift variant and two missense variants of NOD2, encoding a member of the Apaf-1/Ced-4 superfamily of apoptosis regulators that is expressed in monocytes. These NOD2 variants alter the structure of either the leucine-rich repeat domain of the protein or the adjacent region. NOD2 activates nuclear factor NF-kB; this activating function is regulated by the carboxy terminal leucine-rich repeat domain, which has an inhibitory role and also acts as an intracellular receptor for components of microbial pathogens. These observations suggest that the NOD2 gene product confers susceptibility to Crohn's disease by altering the recognition of these components and/or by over-activating NF-kB in monocytes, thus documenting a molecular model for the pathogenic mechanism of Crohn's disease that can now be further investigated. PMID- 11385577 TI - A frameshift mutation in NOD2 associated with susceptibility to Crohn's disease. AB - Crohn's disease is a chronic inflammatory disorder of the gastrointestinal tract, which is thought to result from the effect of environmental factors in a genetically predisposed host. A gene location in the pericentromeric region of chromosome 16, IBD1, that contributes to susceptibility to Crohn's disease has been established through multiple linkage studies, but the specific gene(s) has not been identified. NOD2, a gene that encodes a protein with homology to plant disease resistance gene products is located in the peak region of linkage on chromosome 16 (ref. 7). Here we show, by using the transmission disequilibium test and case-control analysis, that a frameshift mutation caused by a cytosine insertion, 3020insC, which is expected to encode a truncated NOD2 protein, is associated with Crohn's disease. Wild-type NOD2 activates nuclear factor NF kappaB, making it responsive to bacterial lipopolysaccharides; however, this induction was deficient in mutant NOD2. These results implicate NOD2 in susceptibility to Crohn's disease, and suggest a link between an innate immune response to bacterial components and development of disease. PMID- 11385578 TI - The zebrafish Nodal signal Squint functions as a morphogen. AB - Secreted morphogens induce distinct cellular responses in a concentration dependent manner and act directly at a distance. The existence of morphogens during mesoderm induction and patterning in vertebrates has been highly controversial, and it remains unknown whether endogenous mesoderm inducers act directly as morphogens, function locally or act through relay mechanisms. Here we test the morphogen properties of Cyclops and Squint-two Nodal-related transforming growth factor-beta signals required for mesoderm formation and patterning in zebrafish. Whereas different levels of both Squint and Cyclops can induce different downstream genes, we find that only Squint can function directly at a distance. These results indicate that Squint acts as a secreted morphogen that does not require a relay mechanism. PMID- 11385579 TI - MOR1 is essential for organizing cortical microtubules in plants. AB - Microtubules orchestrate cell division and morphogenesis, but how they disassemble and reappear at different subcellular locations is unknown. Microtubule organizing centres are thought to have an important role, but in higher plants microtubules assemble in ordered configurations even though microtubule organizing centres are inconspicuous or absent. Plant cells generate highly organized microtubule arrays that coordinate mitosis, cytokinesis and expansion. Inhibiting microtubule assembly prevents chromosome separation, blocks cell division and impairs growth polarity. Microtubules are essential for the formation of cell walls, through an array of plasma-membrane-associated cortical microtubules whose control mechanisms are unknown. Using a genetic strategy to identify microtubule organizing factors in Arabidopsis thaliana, we isolated temperature-sensitive mutant alleles of the MICROTUBULE ORGANIZATION 1 (MOR1) gene. Here we show that MOR1 is the plant version of an ancient family of microtubule-associated proteins. Point mutations that substitute single amino acid residues in an amino-terminal HEAT repeat impart reversible temperature dependent cortical microtubule disruption, showing that MOR1 is essential for cortical microtubule organization. PMID- 11385580 TI - Metastasis suppressor gene KiSS-1 encodes peptide ligand of a G-protein-coupled receptor. AB - Metastasis is a major cause of death in cancer patients and involves a multistep process including detachment of cancer cells from a primary cancer, invasion of surrounding tissue, spread through circulation, re-invasion and proliferation in distant organs. KiSS-1 is a human metastasis suppressor gene, that suppresses metastases of human melanomas and breast carcinomas without affecting tumorigenicity. However, its gene product and functional mechanisms have not been elucidated. Here we show that KiSS-1 (refs 1, 4) encodes a carboxy-terminally amidated peptide with 54 amino-acid residues, which we have isolated from human placenta as the endogenous ligand of an orphan G-protein-coupled receptor (hOT7T175) and have named 'metastin'. Metastin inhibits chemotaxis and invasion of hOT7T175-transfected CHO cells in vitro and attenuates pulmonary metastasis of hOT7T175-transfected B16-BL6 melanomas in vivo. The results suggest possible mechanisms of action for KiSS-1 and a potential new therapeutic approach. PMID- 11385585 TI - Heavy metal contents in some macrofungi collected in the northwestern part of Turkey. AB - Eight metal contents were determined in wild growing macrofungi collected from two different areas in Balikesir and Akhisar (NW Turkey). The analysis has been done by atomic absorption spectrophotometry in 179 samples of 16 species. Pb was accumulated extensively by G. sessile and L. cinerascens in both areas. Concentration of Cd was significantly high in A. pseudopratensis from background area and M. excissa from near road area. The high concentrations of Fe and Zn were found in M. ramealis and L. cinerascens for both areas. The mean concentration of Cu was high in V. pratense from background area and C. butyracea var. butyracea from near road area. The highest mean concentrations of Co and Mn for both areas were seen in G. sessile and M. excissa, respectively. A high Ni concentration was found in G. sessile and H. hypothejus from background area. We found no considerably significant differences in the concentrations of all studied elements between two areas. PMID- 11385586 TI - Predictions of sediment toxicity using consensus-based freshwater sediment quality guidelines. AB - The objectives of this study were to compare approaches for evaluating the combined effects of chemical mixtures on the toxicity in field-collected sediments and to evaluate the ability of consensus-based probable effect concentrations (PECs) to predict toxicity in a freshwater database on both a national and regional geographic basis. A database was developed from 92 published reports, which included a total of 1,657 samples with high-quality matching sediment toxicity and chemistry data from across North America. The database was comprised primarily of 10- to 14-day or 28- to 42-day toxicity tests with the amphipod Hyalella azteca (designated as the HA10 or HA28 tests) and 10- to 14-day toxicity tests with the midges Chironomus tentans or C. riparius (designated as the CS10 test). Mean PEC quotients were calculated to provide an overall measure of chemical contamination and to support an evaluation of the combined effects of multiple contaminants in sediments. There was an overall increase in the incidence of toxicity with an increase in the mean quotients in all three tests. A consistent increase in the toxicity in all three tests occurred at a mean quotient > 0.5, however, the overall incidence of toxicity was greater in the HA28 test compared to the short-term tests. The longer-term tests, in which survival and growth are measured, tend to be more sensitive than the shorter-term tests, with acute to chronic ratios on the order of six indicated for H. azteca. Different patterns were observed among the various procedures used to calculate mean quotients. For example, in the HA28 test, a relatively abrupt increase in toxicity was associated with elevated polychlorinated biphenyls (PCBs) alone or with elevated polycyclic aromatic hydrocarbons (PAHs) alone, compared to the pattern of a gradual increase in toxicity observed with quotients calculated using a combination of metals, PAHs, and PCBs. These analyses indicate that the different patterns in toxicity may be the result of unique chemical signals associated with individual contaminants in samples. Though mean quotients can be used to classify samples as toxic or nontoxic, individual quotients might be useful in helping identify substances that may be causing or substantially contributing to the observed toxicity. An increase in the incidence of toxicity was observed with increasing mean quotients within most of the regions, basins, and areas in North America for all three toxicity tests. The results of these analyses indicate that the consensus-based PECs can be used to reliably predict toxicity of sediments on both a regional and national basis. PMID- 11385587 TI - Induction of marine mollusc stress proteins by chemical or physical stress. AB - The cellular stress responses of most organisms in part involve the induction of a class of proteins called heat shock or stress proteins (HSPs) as a result of damage to existing proteins. Cellular proteins can be damaged by chemical exposures known to induce various HSPs. In these experiments, we examine the HSP responses of mussel (Mytilus galloprovincialis) and abalone (Haliotis rufescens) tissues to both thermal and chemical exposures. HSP70 isoforms, HSP60, and HSP90 all show varying induction capabilities. The results demonstrate that the extent of stress exposure as both a time- and dose-dependent phenomena can be ascertained by examining changes in mollusc HSP protein levels. We also examined the relationship between HSP induction and levels of a mussel cytochrome P450 (CYP4Y1) mRNA in dose-response experiments with the products of biologically degraded weathered crude oil. The increases in HSP70 isoforms and HSP90 were correlated with decreases in CYP4Y1 expression levels in a dose-dependent manner. HSP responses may therefore be a valuable part of a suite of biomarkers in biomonitoring for hydrocarbon exposures in nearshore environments. PMID- 11385588 TI - The geographic distribution of population health and contaminant body burden in gulf of Mexico oysters. AB - As part of NOAA's National Status and Trends Mussel Watch Program, oysters were sampled along the Gulf of Mexico coast each winter from 1986 to 1993 (The present analysis deals with 1986-1993 Mussel Watch data; the Mussel Watch project itself continues at this printing) and analyzed for trace metal, polynuclear aromatic hydrocarbons and pesticide body burden, plus a series of biological variables designed to assess population status and health. We identified contaminant and biological variables in which large-scale spatial processes played an important role in establishing population values by examining the likelihood that neighboring bays tended to have populations with body burdens or population attributes more similar than expected by chance. Local or watershed-dependent factors, such as land use and freshwater inflow, are important in controlling the bay-to-bay variation in body burden in most contaminants. However, the bay-to-bay variations in body burden of some metals (As, Cd, Hg, Ni, Se) appear to be principally influenced by larger-scale climatic factors. These metals and the biological variable shell length demonstrated a strong degree of similarity between bays over a large regional area reminiscent of the pattern shown by climatic factors, such as temperature and precipitation. In contrast, among the organics, none of the PAHs showed even a moderately strong climatic signal. Among the pesticides, only two did (dieldrin, total DDTs). These pesticides and the biological variables, reproductive stage and Perkinsus marinus prevalence and infection intensity, had spatial patterns that suggested both a local and a regional influence to their geographic distributions. This same pattern is exhibited by freshwater runoff. Metal contaminants also behaved distinctively compared to organics in the temporal influence of climate in establishing the interannual variability in body burden. For the organics, trends in interannual variability were strongly influenced by climate, whereas spatial trends were not. In contrast, most metals were unaffected by climatic forcing both spatially and temporally. However, all of the metals having a spatial pattern strongly influenced by climate (As, Cd, Hg, Ni, Se) also exhibited interannual variations related to variations in climate. PMID- 11385589 TI - Effects of methylmercury on ontogeny of prey capture ability and growth in three populations of larval Fundulus heteroclitus. AB - We used three populations of mummichogs (Fundulus heteroclitus), one from a polluted site (Piles Creek [PC], New Jersey) and two from cleaner sites (Tuckerton [TK], New Jersey, and East Hampton [EH], New York), to study (1) whether embryonic, embryonic plus larval, or larval exposure to methylmercury (MeHg) altered larval prey capture ability and growth; and (2) whether there were differences in tolerance to MeHg-induced behavioral changes among the three populations. Eggs and sperm were obtained from mummichogs captured in the field, and their embryos and larvae were kept in clean sea water or MeHg solution (5, 10 microg/L). Larvae were then tested regularly for prey capture rates and prey capture efficiencies, and their lengths were measured. Embryonic exposure to MeHg induced transitory and recoverable impairments in larval prey capture ability, whereas larval exposure alone was relatively ineffective. When both embryos and larvae were treated, larval prey capture ability was affected at a lower concentration and a wider range of larval ages. In terms of growth and prey capture ability, response of larvae to embryonic or larval or exposure to both stages to MeHg varied with populations. TK fish were the most tolerant with respect to behavioral changes but were the most sensitive to MeHg in reduction of growth. EH fish were the most sensitive whenever embryos were treated, and PC fish were the most vulnerable after larval exposure. The population differences in response to MeHg intoxication may be due to pollution related factors or differences in behavioral-related genetic factors. PMID- 11385590 TI - Effects of acute handling stress on whitefish Coregonus lavaretus after prolonged exposure to biologically treated and untreated bleached kraft mill effluent. AB - Exposure of fish to water of impaired quality has been shown to disrupt the function of the hypothalamo-pituitary-interrenal (HPI) axis and alter the interpretation of data from field studies due to the varying effects of handling and delayed sampling on exposed and reference animals. In the present study, juvenile whitefish, Coregonus lavaretus, were exposed for 6 weeks to diluted (4 8%) untreated and biologically treated bleached kraft mill effluent (BKME) and their response to acute handling was investigated. Liver microsomal EROD activity and glycogen phosphorylase (GPase) activity, in addition to gill Na+-K+-ATPase activity, and blood hemoglobin and hematocrit levels were increased in whitefish exposed for 6 weeks to untreated BKME, whereas those exposed to treated BKME exhibited increased blood hemoglobin and red blood cell K+ concentrations. Both handling procedures, exposure to a shallow water (10 cm, 5 min) and to an air challenge (10 s air/10 s water/30 s air/10 s water/10 s air), resulted in acute physiological stress, as recorded after 5-, 60-, and 120-min recovery periods. Following air exposure, the levels of plasma cortisol, blood glucose, hemoglobin, and hematocrit as well as the liver GPase activity were increased, and liver glycogen concentration decreased in control fish. These responses were attenuated in fish exposed to untreated or treated BKME. Plasma estradiol and testosterone levels were not affected by the BKME exposures or by the air challenge. Handling also resulted in attenuated EROD induction in fish exposed to untreated BKME. According to the present findings, the sensitivity of some widely used cellular and physiological variables may be improved by time-dependent standardization when interpreting data obtained following delayed sampling. PMID- 11385591 TI - Porphyrin levels in excreta of sea birds of the Chilean coasts as nondestructive biomarker of exposure to environmental pollutants. AB - In this preliminary study on sea birds we propose the use of porphyrins in excreta as a biomarker of exposure to contaminants. Samples of excreta were obtained from colonies of brown pelicans (Pelecanus occidentalis thagus), neotropic cormorants (Phalacrocorax olivaceus), and kelp gulls (Larus dominicanus) in three areas of the south coast of Chile with different human impact (Tubul, Talcahuano, and Valdivia). They were analyzed for porphyrin content (copro-, uro-, and protoporphyrins and total porphyrins) by a rapid fluorimetric method and by HPLC. The main outcomes of the study were: (a) kelp gulls and neotropic cormorants living in areas with high human impact showed a clear capacity to accumulate and eliminate porphyrins in the excreta; (b) species related accumulation capacities are likely, as shown by the different levels found in different species living in the same area; (c) the porphyrin profile obtained by fluorimetry and HPLC showed a higher percentage of protoporphyrin than the other porphyrins; (d) although the fluorimetric method of Grandchamp is semiquantitative, it was found to be sensitive enough to detect differences in samples from field studies. The positive results of this preliminary study make it possible to propose this nondestructive method for a variety of field applications. PMID- 11385592 TI - Organochlorine contaminant exposure and reproductive success of black-crowned night-herons (Nycticorax nycticorax) nesting in Baltimore harbor, Maryland. AB - The declining size of the Baltimore Harbor black-crowned night-heron (Nycticorax nycticorax) colony has been hypothesized to be linked to polychlorinated biphenyl (PCB) exposure. In 1998, a "sample egg" was collected from 65 black-crowned night heron nests (each containing > or = three eggs) for contaminant analysis, and the remaining eggs in these 65 nests, plus four two-egg nests, were monitored for hatching and fledging success. Eggs were also collected from 12 nests at Holland Island, a reference site in southern Chesapeake Bay. Samples were analyzed for 26 organochlorine pesticides and metabolities and 145 PCB congeners. Pesticide and metabolite concentrations, including p,p'-DDE, were well below thresholds associated with adverse reproductive effects at both sites. Average concentration of total PCBs, 12 Ah receptor-active PCB congeners, and toxic equivalents in eggs from Baltimore Harbor were greater (up to 35-fold) than that observed in Holland Island samples. Overall nest success at the Baltimore Harbor heronry was estimated by the Mayfield method to be 0.74, and the mean number of young fledged/hen was 2.05, which is within published productivity estimates for maintaining a stable black-crowned night-heron population. Using logistic regression, no significant relationships were found between organochlorine contaminant concentrations in sample eggs and hatching, fledging, or overall reproductive success. Processes other than poor reproduction (e.g., low postfledging survival, emigration, habitat degradation) may be responsible for the declining size of the Baltimore Harbor colony. PMID- 11385593 TI - Protection of sodium arsenite-induced ovarian toxicity by coadministration of L ascorbate (vitamin C) in mature wistar strain rat. AB - Arsenic, a major water pollutant in India, produces toxic effects on female reproductive system in rodent models at the dose available in drinking water in arsenic-intoxicated zones. This study examines the coadministration of L ascorbate (vitamin C) on ovarian steroidogenesis, plasma levels of gonadotrophins, brain monoamines, and ovarian as well as uterine peroxidase activities in sodium arsenite-treated rats. After sodium arsenite treatment, relative ovarian and uterine weights, ovarian Delta5-3beta-HSD and 17beta-HSD activities, plasma levels of gonadotrophins, norepinephrine levels in midbrain and diencephalon, and the activities of peroxidase in ovary and uterus were decreased significantly. On the other hand, serotonin levels in midbrain and diencephalon were increased significantly 28 days after sodium arsenite treatment at the dose of 0.4 ppm/100 g body weight/rat/day. All these parameters were protected significantly and in most cases were unchanged from control level when L-ascorbate at 25 mg/100 g body weight/rat/day was coadministered orally with sodium arsenite. This cotreatment of L-ascorbate with sodium arsenite also restored the estrous cycle in a regular manner. We concluded that L-ascorbate plays a pivotal role in maintaining normal ovarian activities and brain monoamines in arsenic-treated rats. PMID- 11385594 TI - Organochlorine pesticides, polychlorinated biphenyls, and butyltin compounds in blubber and livers of stranded California sea lions, elephant seals, and harbor seals from coastal California, USA. AB - Concentrations of polychlorinated biphenyls (PCBs), DDTs (p,p'-DDE, p,p'-DDD, p,p'-DDT), chlordanes (CHLs; cis-chlordane, cis-nonachlor, trans-nonachlor, and oxychlordane), hexachlorocyclohexane isomers (HCHs), hexachlorobenzene (HCB), tris(4-chlorophenyl)methane (TCPMe), tris(4-chlorophenyl)methanol (TCPMOH), and mono- (MBT), di-(DBT), and tri-butyltin (TBT) were determined in blubber and livers of 15 California sea lions (Zalophus californianus), 6 northern elephant seals (Mirounga augustirostris), and 10 harbor seals (Phoca vitulina) found stranded along the coasts of California, USA, during 1991-1997. Among the organochlorines analyzed, DDTs were predominant, followed in decreasing order by PCBs, CHLs, TCPMe, TCPMOH, HCHs, and HCB. The greatest concentrations of organochlorines were found in California sea lions. The highest DDT and PCB concentrations found in the blubber of California sea lions were 2,900 and 1,300 microg/g, lipid weight, respectively. Concentrations of TCPMe and TCPMOH in California sea lions were correlated significantly with DDT concentrations. Concentration ratios of various organochlorines in harbor seal livers were different from those in California sea lions and elephant seals, which suggested that the sources of exposure of harbor seals to organochlorines were different from those in the other two species. Concentrations of butyltin compounds in livers of pinniped species ranged from 2 to 99 ng/g, wet weight, which were less than those observed in cetaceans and in California sea otters. PMID- 11385595 TI - Cadmium accumulation and distribution in slaughtered horse kidneys from the Argentine central region. AB - In this paper we report the results of surveys conducted in Argentina between 1997 and 1998 to know the Cd concentrations in kidney from horses of different age, sex, and origin. Cd in renal cortex and medulla was positively correlated, and higher concentrations in the cortex were found. No significant differences between values from left and right kidneys of the same animal were found. An increase in Cd levels with age of animals were observed, and no sex incidence was verified in renal Cd composition. No detectable residues were found in the fetuses tested. Levels observed in Argentine equines (n = 102) ranged from 4.3 to 83.8 microg Cd/g in kidney cortex, lower than those reported by other authors but higher than the action levels proposed by Argentine meat tissue species regulations. Therefore, the study remarks on the convenience of monitoring Cd in Argentine equines and gives relevant information for regulatory purposes to consider the limitation in the use of horse kidney as food for either human or animals. PMID- 11385596 TI - Evaluation of immune parameters in propanil-exposed farm families. AB - The rice herbicide propanil induces alterations in the mouse immune system, causing significant decreases in T cell-dependent and T cell-independent antibody responses. This postemergent herbicide is used extensively in rice production in the Mississippi River delta region of the southern United States. The aerial application and airborne drift of propanil may pose health concerns to exposed farm families living adjacent to sprayed rice fields. To determine if aerial spraying of propanil increases risks of altered immune responses in families bordering rice fields, immune parameters were assessed during a 2-year study. Families living within 100 yards of rice fields were compared in a case control study to farm families whose homes exceeded 1 mile from any rice field. Blood was analyzed in adults (n = 56) and children (n = 52) at three time intervals: (1) preseason, prior to propanil application; (2) 5-7 days after aerial application of propanil to rice fields; and (3) postseason, following harvest. Exposed adults and children were compared with controls for a number of immune parameters. Total cell count and the percentage of various lymphocytes (T cells, B cells, CD4+ helper cells, and CD8+ suppressor cells) and natural killer (NK) cells, mitogen induced cell proliferation, cytokine (IL-2+) production, and NK cell function were assessed. A comparison of immune function between exposed and nonexposed farm families showed no significant differences, possibly related to propanil exposure. However, some immune test parameters changed as a function of season rather than propanil exposure. The data indicate that individuals living next to rice fields are not at increased risk of altered immune function due to propanil exposure. PMID- 11385597 TI - Propanil (3,4-dichloropropionanilide) particulate concentrations within and near the residences of families living adjacent to aerially sprayed rice fields. AB - Propanil is widely used as a postemergence herbicide in rice. Because it is typically applied aerially, there is a potential for propanil to drift into and around homes of those living adjacent to rice fields. Propanil has been shown to be immunotoxic in rodent models. The objective of this study was to measure the levels of propanil to which families living adjacent to aerially sprayed rice fields may be exposed. Air levels were sampled by actively and passively collecting propanil in and around the homes of volunteer families living in close proximity to rice fields sprayed with propanil. Homes ranged from 73 m to 113 m from treated rice fields. Sampling was conducted in the home, adjacent to the home (within 5 m of the home), 30 m from the home, and at the edge of the rice field. Concentrations were determined via gas chromatography/mass spectroscopy. Propanil levels adjacent to the homes ranged from nondetectable to 1,106.4 microg per 400 cm2 collection surface (2.0 microg detection limit). Wind direction and wind velocity were the primary determinants of propanil drift. At sites where the prevailing wind was blowing away from the home, no propanil was detected except at the edge of the field. Distance from the edge of the rice field also influenced the amount of drift with higher levels measured at 30 m from the house than adjacent to the house. No propanil vapor was detected on absorbent media sampled in and around the homes. The results indicate that individuals living adjacent to rice fields aerially sprayed with propanil are potentially exposed to variable amounts of propanil, and wind speed and direction are the most important factors that influence the concentration of aerially applied pesticide. PMID- 11385598 TI - Proteomic comparison of human and great ape blood plasma reveals conserved glycosylation and differences in thyroid hormone metabolism. AB - Most blood plasma proteins are glycosylated. These glycoproteins typically carry sialic acid-bearing sugar chains, which can modify the observed molecular weights and isoelectric points of those proteins during electrophoretic analyses. To explore changes in protein expression and glycosylation that occurred during great ape and human evolution, we subjected multiple blood plasma samples from all these species to high-resolution proteomic analysis. We found very few species-specific differences, indicating a remarkable degree of conservation of plasma protein expression and glycosylation during approximately 12 million years of evolution. A few lineage-specific differences in protein migration were noted among the great apes. The only obvious differences between humans and all great apes were an apparent decrease in transthyretin (prealbumin) and a change in haptoglobin isoforms (the latter was predictable from prior genetic studies). Quantitative studies of transthyretin in samples of blood plasma (synthesized primarily by the liver) and of cerebrospinal fluid (synthesized locally by the choroid plexus of the brain) confirmed approximately 2-fold higher levels in chimpanzees compared to humans. Since transthyretin binds thyroid hormones, we next compared plasma thyroid hormone parameters between humans and chimpanzees. The results indicate significant differences in the status of thyroid hormone metabolism, which represent the first known endocrine difference between these species. Notably, thyroid hormones are known to play major roles in the development, differentiation, and metabolism of many organs and tissues, including the brain and the cranium. Also, transthyretin is known to be the major carrier of thyroid hormone in the cerebrospinal fluid, likely regulating delivery of this hormone to the brain. A potential secondary difference in retinoid (vitamin A) metabolism is also noted. The implications of these findings for explaining unique features of human evolution are discussed. PMID- 11385599 TI - Individual variation in the growth of captive infant gorillas. AB - Serial anthropometric data were obtained during the first year of life of six nursery-reared infant gorillas in the Columbus (Ohio) Zoo. Two of the infants are likely to be monozygotic twins as determined by DNA analysis. Growth curves were fitted to serial measures of cephalo-thoracic-abdominal length, arm length, leg length, head circumference, upper arm circumference, and weight from each gorilla, to describe individual patterns of variation in skeletal growth and body composition. Growth in skeletal measures tended to be curvilinear to varying degrees over the first year of life. Body composition varied more than skeletal measures throughout the first year as a consequence of individual health status. Individual growth and body composition variations appear to reflect both genetic and environmental influences in this small sample of captive infant gorillas. PMID- 11385600 TI - Genetic relationships between Amerindian populations of Argentina. AB - A total of 495 individuals from five different Argentinian tribes was examined for variation in 23 blood group and protein genetic systems, and the results were integrated with previous data on some of these systems. These tribes generally present RH * R1, PGM1 * 1, and ACP * A frequencies lower and RH * R2, ESD * 1, and GLO * 1 prevalences higher than those observed in other South American Indian groups. Earlier studies with mitochondrial DNA showed that haplogroup A was present in low frequencies in these tribes, but haplogroup B showed a high prevalence among the Mataco. Average heterozygosities are very similar in the five tribes, while estimates of non-Indian ancestry are generally low. Both the blood group and protein, as well as the mtDNA data sets, divide the five tribes into two groups, and the relationships obtained with the blood group and protein systems are exactly those expected on the basis of geography and language. However, the topology obtained with the mtDNA results was different, possibly due to sampling effects or diverse patterns of exchange between the groups related to sex. PMID- 11385601 TI - DNA diversity and population admixture in Anatolia. AB - The Turkic language was introduced in Anatolia at the start of this millennium, by nomadic Turkmen groups from Central Asia. Whether that cultural transition also had significant population-genetics consequences is not fully understood. Three nuclear microsatellite loci, the hypervariable region I of the mitochondrial genome, six microsatellite loci of the Y chromosome, and one Alu insertion (YAP) were amplified and typed in 118 individuals from four populations of Anatolia. For each locus, the number of chromosomes considered varied between 51-200. Genetic variation was large within samples, and much less so between them. The contribution of Central Asian genes to the current Anatolian gene pool was quantified using three different methods, considering for comparison populations of Mediterranean Europe, and Turkic-speaking populations of Central Asia. The most reliable estimates suggest roughly 30% Central Asian admixture for both mitochondrial and Y-chromosome loci. That (admittedly approximate) figure is compatible both with a substantial immigration accompanying the arrival of the Turkmen armies (which is not historically documented), and with continuous gene flow from Asia into Anatolia, at a rate of 1% for 40 generations. Because a military invasion is expected to more deeply affect the male gene pool, similar estimates of admixture for female- and male-transmitted traits are easier to reconcile with continuous migratory contacts between Anatolia and its Asian neighbors, perhaps facilitated by the disappearance of a linguistic barrier between them. PMID- 11385602 TI - Nonhuman primate hybridization and the taxonomic status of Neanderthals. AB - The present study examines the taxonomic status of Middle Paleolithic Neanderthals by comparing their observed minimum genetic divergence from Upper Paleolithic modern humans in Europe with that observed between macaque species from Sulawesi that are known to hybridize and fully intergrade in the wild. The genetic divergence, and differentiation between Neanderthals and Upper Paleolithic modern humans, as indicated by pairwise minimum genetic distances and F(ST) values calculated from the estimated minimum genetic relationship (R) matrix derived from craniometric data, are significantly greater than those observed both between hybridizing and noninterbreeding Sulawesi macaque species, suggesting that mate recognition and the possibility of gene flow between Neanderthals and Upper Paleolithic modern humans might have been greatly reduced. These results support a species-level taxonomic distinction for the Neanderthals as suggested by proponents of the replacement model. Furthermore, assumptions regarding the monophyletic origin of modern humans from outside Europe are likely valid. PMID- 11385603 TI - Testing the taxonomic integrity of Paranthropus boisei sensu stricto. AB - The craniodental hypodigm of Paranthropus boisei sensu stricto is morphologically distinctive, but it has been suggested that the substantial variation in mandibular and dental size in that hypodigm may exceed that which is reasonable to subsume within a single hominin species. In this study, Fligner and Killeen, coefficient of variation (CV)-based and average taxonomic distance (ATD)-based bootstrap tests, were used to compare variation in size and shape of the mandibular corpus remains attributed to P. boisei s.s. with the variation observed in samples of great apes and modern humans. The degree of size variation in the P. boisei s.s. mandibular hypodigm is never observed in human and chimpanzee samples, is rare in gorillas, but is not uncommon in orangutans. However, the shape variation in the fossil group is comparable to the variation in the extant reference groups. Although the size variation in P. boisei s.s. is substantial, it is exaggerated by the effects of taphonomy. The small mandibles are more often abraded, whereas the large mandibles are more likely to have been infiltrated with matrix. On the basis of the results of this investigation of the mandibular corpus, there are no grounds for rejecting the "single-species" hypothesis for P. boisei s.s. When Sokal and Braumann's adjusted CV values were used to predict the index of sexual dimorphism (ISD) for the P. boisei s.s., despite the substantial geological time embraced by the mandibular corpus hypodigm, the predicted value of lnISD, when corrected for taphonomic factors, is comparable to the sexual dimorphism observed within Gorilla. PMID- 11385604 TI - Sexually dimorphic mandibular morphology in the first few years of life. AB - Sex differences in the youngest skeletons are very subtle, and any method that can separate males and females significantly better than chance will be of value. Compounding the problem is a paucity of immature skeletons of documented age and sex. In 1992, S.R.L. examined 62 juvenile mandibles of white and black South Africans of known age and sex (from birth to 19 years) from the Dart Collection to determine if the sexes could be differentiated by morphologic traits. By age 6 years, adult chin shapes were already recognizable. Prior to that age, differences were observed in the shape of the inferior border of the symphysis and outline of the body. The male chin base extends steeply downward relative to the adjacent body, coming to a point or squaring off at the symphysis. In females, the symphysis descends gradually to a more rounded base, and even when pointed, the transition is not abrupt. On the outer border of the corpus, the sides diverge sharply to form a ?_/ shape from a roughly horizontal anterior region in males, while the female contour is rounded, reflecting the smoothly curved transition from front to sides. These traits were manifest from the eruption of the central incisors until about 4 years of age. The features were tested on all 19 Dart Collection mandibles in that age range. Average accuracy for three different testers was 81%, and males were consistently identified more accurately than females. This new method was then tested on a known sex sample of 11 individuals from 0 to 7 years of age. These included CT scans of 9 French children and the remains of 2 South African black forensic cases. Sexing accuracy was 82% (9/11). The only two missexed cases were both female and over age 6 years. In conclusion, the results of this study indicate that it is possible to determine the sex of very young mandibles. The new sexually dimorphic morphologic configurations introduced here have demonstrated repeatable discrimination with the highest level of accuracy (81%) reported and tested for this age group. Preliminary research indicates that both the male and female shapes are clearly recognizable in archaeologic and premodern hominids as well as chimpanzees. PMID- 11385605 TI - Craniodental body mass estimators in the dwarf bushbaby (Galagoides). AB - This study reports data on 17 craniodental body mass estimators in a sample (n = 38) of dwarf galagos (Galagoides). Correlation coefficients (r) range from a high of 0.64 for bizygomatic breadth and body mass to a low of 0.10 for M(3) length and body mass. Of the 17 variables studied, 7 exhibit significant (P < 0.05) correlation coefficients, with 5 of the 7 being multitooth (i.e., tooth row) or cranial variables. In contrast to the correlation coefficients of greater than 0.90 (e.g., Martin [1980] Z Morphol Anthropol 71:115-124; Steudel [1981] Int J Primatol 2:81-90; Gingerich et al. [1982] Am J Phys Anthropol 58:81-100; Conroy [1987] Int J Primatol 8:115-137) published for higher taxonomic level analyses (i.e., all-primate or prosimian) for many of the same variables studied here, the current data indicate weaker relationships when analyzed at the generic level. Possible explanations for the contrast in correlation coefficients between the current and many previous studies include the following: 1) individual variation due to a geographically dispersed sample, 2) individual body mass fluctuations due to seasonal food availability, and 3) individual variation within the sample due to variation in life-history parameters. Because the overall size range of the individuals in a specific or generic level analysis is smaller than that in an ordinal or subordinal sample, the individual variation normally masked when using species means represents a larger proportion of the total variation in a more limited sample. This may then be a cause of these weaker correlations. PMID- 11385606 TI - Letter to the editor: response to a dismal view. PMID- 11385607 TI - Control of T cell hyperactivation in IL-2-deficient mice by CD4(+)CD25(-) and CD4(+)CD25(+) T cells: evidence for two distinct regulatory mechanisms. AB - In IL-2-deficient mice, antigen-activated CD4 T cells accumulate and cause lethal immune pathology. Wild-type cells of hematopoietic origin present in the same animal are able to prevent this hyperactivation of T cells, but the mechanisms and cells controlling the IL-2-deficient cells are unknown. Here we show that IL 2(-) CD4 cells with an ovalbumin-specific transgenic TCR (IL-2(-) OVAtg) undergo both clonal expansion and clonal contraction when transferred to euthymic recipients and challenged with antigen, but continuously expand in athymic hosts. Cotransfer of wild-type CD4 T cells prevents the accumulation of IL-2-deficient cells. On the residual IL-2(-) TCRtg cells CD69 and CD25 are up-regulated, suggesting that activation per se is not suppressed and that the cells had received an IL-2 signal. Since IL-2 is able to restore the defective antigen induced cell death (AICD) of IL-2-deficient T cells in vitro, paracrine IL-2 provided by the wild-type CD4 cells may thus be able to allow clonal contraction of IL-2-deficient cells also in vivo. Interestingly however, regulatory CD4(+)CD25(+) cells also efficiently contain the clone size of antigen-stimulated IL-2-deficient T cells. Since CD4(+)CD25(+) cells do not produce IL-2, this suggests a mechanism of suppression distinct from paracrine IL-2 delivery. In keeping with this, the residual IL-2(-) TCRtg cells recovered after cotransfer of regulatory CD4(+)CD25(+) cells do not show increased CD25 or CD69 expression, suggesting that they had not received paracrine IL-2 and that clonal containment occurred at the level of initial activation rather than clonal contraction by AICD. IL-2 deficiency therefore may upset T cell homeostasis by two distinct mechanisms: the failure to program expanding T cells for apoptosis, and the failure to generate functional CD4(+)CD25(+) regulatory cells. PMID- 11385608 TI - Phenotype and function of human dendritic cells derived from M-DC8(+) monocytes. AB - Dendritic cells (DC) generated from peripheral blood monocytes have been used with promising results as a new approach for the immunotherapy of cancer. However, at least four different subpopulations of peripheral blood monocytes have been recognized and their contribution to the generation of functional DC is not known. Recently, the monoclonal antibody M-DC8 has been shown to react with 0.2 - 1 % of blood leukocytes. We have identified M-DC8(+) cells as monocytes which represent about 40 % of CD14(low)CD16(+) monocytes. Similar to M-DC8(-) monocytes, they develop in the presence of GM-CSF and IL-4 into a very homogenous population of cells with DC phenotype and function. M-DC8(+) DC show on average a twofold higher expression of HLA class I and class II molecules than M-DC8(-) DC. These DC produce IL-12p75 both in response to LPS and to CD40 ligation. M-DC8(+) DC induced a strong Th1 immune response and were two to four old more potent than M-DC8(-) DC for the priming of cord blood T cells. M-DC8(+) monocytes can be used as a source of very potent dendritic cells with the potential to significantly improve the efficacy of DC-based immunotherapies. PMID- 11385609 TI - NK cell-mediated lysis of autologous antigen-presenting cells is triggered by the engagement of the phosphatidylinositol 3-kinase upon ligation of the natural cytotoxicity receptors NKp30 and NKp46. AB - Interleukin-2 (IL-2)-activated polyclonal or clonal NK cells lysed autologous antigen presenting cells (APC) through the engagement of the natural cytotoxicity receptors (NCR) NKp30 and NKp46. NK cell-mediated cytolysis of APC correlated with the surface density of these NCR. Indeed, NK cell clones bearing low amounts of NKp30 and NKp46 did not lyse autologous APC, whereas NK cell clones with bright expression of these NCR efficiently killed autologous APC. Upon masking of NKp30 or NKp46 by specific monoclonal antibodies a strong reduction (by 50%) of APC lysis could be detected and the complete inhibition was achieved by the simultaneous masking of these NCR. Interestingly, NK cell-mediated APC lysis was impaired by the phosphatidylinositol 3-kinase (PI-3 K) inhibitors LY294002 or wortmannin. Similarly, these drugs strongly reduced NK cell activation triggered by NKp30 or NKp46 in a re-directed killing assay as well as the activation of Akt/PKB, substrate of PI-3 K, induced by the engagement of these receptors. Altogether, these findings strongly suggest that NCR are responsible for the killing of autologous APC through the activation of PI-3 K. PMID- 11385610 TI - Cartilage-specific autoimmunity in rheumatoid arthritis: characterization of a triple helical B cell epitope in the integrin-binding-domain of collagen type II. AB - Cartilage-specific proteins are considered potential autoantigens that could continuously fuel autoimmune responses directed to the joints in rheumatoid arthritis (RA). Using recombinant chimeric collagen type II we have identified one major type II collagen (CII) epitope (denoted U1) recognized by RA sera. The U1 epitope is a triple helical structure formed by 11 amino acids (triple helical position 494-504) and colocalizes with the recently described alpha1beta1/alpha2beta1 integrin binding site. It is a major epitope, found in 14/22 RA sera positive for antibodies to CII. One individual could be followed for a long time and the results showed that IgG antibodies specific for the U1 epitope were maintained along the chronic disease course but suppressed during periods of cyclosporin A and anti-CD4 treatment. We also found that the U1 epitope was recognized in rats susceptible to collagen-induced arthritis. A monoclonal autoantibody (mAb 126.30) was raised from DA rats, which bound the same epitope. The antibodies bound the cartilage in vivo showing that the epitope is exposed to the immune system for immune complex formation in the intact joint. PMID- 11385611 TI - Immune evasion of Borrelia burgdorferi by acquisition of human complement regulators FHL-1/reconectin and Factor H. AB - To understand immune evasion mechanisms of Borrelia burgdorferi we compared serum resistant B. afzelii and serum-sensitive B. garinii isolates for their capacity toacquire human complement regulators. Here we demonstrate that the two borrelial genospecies show different binding of the two important human complement regulators, FHL-1/reconectin and Factor H. All serum-resistant B. afzelii isolates bound FHL-1/reconectin and also Factor H, and all analyzed serum sensitive B. garinii isolates showed no or a significantly lower binding activity. Using recombinant deletion mutants, the binding domains were localized to the C terminus of FHL-1/reconectin to short consensus repeats 5-7. The borrelial binding proteins were located in the surface of the bacteria as demonstrated by immunofluorescence staining of intact, serum-exposed bacteria and by enrichment of outer membrane proteins. The surface-attached complement regulators maintained complement regulatory activity as demonstrated in a cofactor assay. By ligand blotting two different borrelial binding proteins were identified that were responsible for the surface attachment of FHL-1/reconectin and Factor H. These borrelial complement regulators acquiring surface proteins (CRASP) were further characterized as either CRASP-1, a 27.5-kDa molecule which preferentially binds FHL-1/reconectin and which was present in all serum resistant borreliae, or CRASP-2, a 20/21-kDa protein which interacts preferentially with Factor H and the expression of which was more restricted, being detected in four of the six isolates analyzed. In summary, we describe a new immune evasion mechanism of B. burgdorferi, as these bacteria acquire human complement regulators to control complement activation on their surface and to prevent formation of toxic activation products. PMID- 11385612 TI - Both CD45R(low) and CD45R(high) "revertant" CD4 memory T cells provide help for memory B cells. AB - During a primary response to thymus dependent antigens, B cells undergo a number of qualitative changes to become memory B cells - processes that require co stimulatory signals and cytokine help from CD4 T cells. The question of whether distinct, antigen-experienced memory CD4 T cells are subsequently needed to program memory B cells into antibody synthesis has not been clearly resolved. Using an adoptive transfer model in which memory but not naive B cells were stimulated, we evaluated CD4 T cell help using lymphocytes obtained from primed or unprimed thymectomized donors and expressing a naive (CD45R(high)) or a memory (CD45R(low)) phenotype. Memory B cells, most of which were committed to the IgG1 (Th2) subclass, could be stimulated to produce antibody using help transferred by the CD45R(high) naive subset of unprimed donors (slow onset of response), the CD45R(low) subset of 7 day primed donors (large, rapid antibody response) or by both the CD45R(low) and the CD45R(high) "revertant" subsets of 6 month primed donors. We found that antigen primed CD45R(low) CD4 T cells reverted (defaulted) with time to a CD45R(high) resting state, a change that was prevented by persisting antigen. The evidence suggests that CD4 memory T cells are partitioned into two different functional states (CD45R(high) and CD45R(low)) and that these determine the characteristics of the memory B cell response in terms of speed, size and longevity. PMID- 11385613 TI - A conserved sequence block in the murine and human T cell receptor Jalpha loci interacts with developmentally regulated nucleoprotein complexes in vitro and associates with GATA-3 and octamer-binding factors in vivo. AB - A highly conserved sequence block (CSB) located in the mouse and human T cell receptor (TCR) Jalpha loci is recognized by tissue-specific factors and up regulates TCR alpha enhancer activity. In this study, the properties of CSB interacting factors were further explored to decipher the function of this cis acting element. Thymocytes corresponding to different developmental stages were found capable of forming differential CSB-nucleoprotein complexes. Pronounced changes in the CSB-complex-forming activity were observed during the transition from double-negative to double-positive thymocytes. Furthermore, we showed that transcription factors Oct-1, Oct-2 and GATA-3 interacted with CSB both in vitro, as evidenced by electrophoretic mobility shift assays, and in vivo, as demonstrated by chromatin immunoprecipitation assays in mouse thymus. Importantly, we also demonstrated that GATA-3 associated in vivo with TCR alpha enhancer, the activity of which is known to be required in regulating chromatin accessibility to the V(D)J recombinase. Thus, CSB may temporally regulate local chromatin structure and help to spread TCR alpha enhancer activity over the entire 70-kb Jalpha locus by forming developmentally regulated CSB-nucleoprotein complexes and by interacting with other cis-regulatory element-nucleoprotein complexes present within the TCR alpha / delta locus. PMID- 11385614 TI - Smad3 and Smad4 mediate transforming growth factor-beta1-induced IgA expression in murine B lymphocytes. AB - Transforming growth factor (TGF)-beta1 is well established as a critical IgA isotype switching factor and Smad molecules have been reported to act as transducers and transcriptional factors in the expression of TGF-beta1-targeted genes. We examined the involvement of Smad proteins in TGF-beta1-induced IgA expression. First, we found that TGF-beta1 significantly increases endogenous germ-line (GL) alpha transcripts by LPS-stimulated CH12.LX.4933 (mu(+)) B lymphoma cells. To investigate its signaling mechanisms, the lymphoma cell line was transfected with pFL3 that contains the TGF-beta-responsive element of the GLalpha promoter, and stimulated with TGF-beta1. Similar to endogenous GLalpha transcripts, TGF-beta1 induces GLalpha promoter activity and overexpression of Smad3 markedly enhances the promoter activity. This activity is further augmented by cotransfected Smad4. On the other hand, Smad7 substantially abrogates the synergistic effect of Smad3/4 onGLalpha promoter activity. In addition, overexpression of Smad3/4 enhances TGF-beta1-induced endogenous GLalpha transcripts in normal spleen B cells. Finally, in the presence of TGF-beta1, overexpression of Smad3/4 selectively increases both surface IgA expression and IgA production. The results from the present study indicate that Smad3, Smad4, and Smad7, at least in part, serve as mediators linking TGF-beta1 to transcriptional regulation of IgA switching related gene and regulation of IgA class switching. PMID- 11385615 TI - CD14(dim)/CD16(bright) monocytes in hemophagocytic lymphohistiocytosis. AB - Hemophagocytic lymphohistiocytosis (HLH) is an extremely rare and highly lethal chronic inflammatory disease, which is mediated by proinflammatory cytokines. In the peripheral blood of a boy suffering from HLH, a chronic expansion of CD14(dim)/CD16(bright) inflammatory monocytes was detected. Compared with CD14(bright) monocytes, their immunophenotype correlated with more mature monocytic cells differentiating to macrophages: they showed lower expression of CD11b, CD64 and CD35. Such CD14(dim)/CD16(bright) monocytes produce the inflammatory cytokines IL-1beta, IL-6 and TNF-alpha. They fit in well with the pathophysiological concept of HLH as an inflammatory state of lymphocytes and of the monocyte/macrophage system. In the presented patient the percentage of these circulating inflammatory monocytes decreased over time during clinical response to immunosuppressive therapy. This finding may indicate that CD14(dim)/CD16(bright) monocytes represented the degree of inflammation in this extremely rare and highly lethal disease. PMID- 11385616 TI - Critical contribution of IFN-gamma and NK cells, but not perforin-mediated cytotoxicity, to anti-metastatic effect of alpha-galactosylceramide. AB - The glycolipid alpha -galactosylceramide (alpha -GalCer), which is presented by CD1d and specifically activates Valpha 14 NKT cells, exerts a potent anti metastatic effect when administered in vivo. In this study, we demonstrated that alpha -GalCer administration led to rapid elimination of NKT cells by apoptosis in the liver and spleen, after they produced IFN-gamma and IL-4. In contrast, a more prolonged secretion of IFN-gamma was observed by liver and splenic NK cells after alpha -GalCer administration. Cytotoxic activity of liver mononuclear cells was not augmented 3h after alpha -GalCer administration, but was increased at 24 h when NKT cells were mostly depleted. The alpha -GalCer-induced cytotoxic activity was abolished in IFN-gamma -deficient and NK cell-depleted mice as well as CD1-deficient mice, suggesting that the alpha -Galcer-induced cytotoxicity was mainly mediated by IFN-gamma -activated NK cells. While the alpha -GalCer-induced cytotoxicity in vitro was mostly perforin dependent, anti-metastatic effect of alpha -GalCer was impaired in NK cell-depleted or IFN-gamma -deficient mice but not in perforin-deficient mice. Collectively, these results indicated that the anti-metastatic effect of alpha -GalCer is mainly mediated by NK cells, which are activated secondarily by IFN-gamma produced by alpha -GalCer-activated NKT cells, in a perforin-independent manner. PMID- 11385617 TI - KIR down-regulation on NK cells is associated with down-regulation of activating receptors and NK cell inactivation. AB - We previously reported that killer cell immunoglobulin-like receptors (KIR) could be down-regulated from the surface of T cells. Here, we show that KIR down regulation is also induced on the surface of natural killer (NK) cells upon ligand binding. Common down-regulation characteristics are found on these two cell types: a slow kinetics and a phenomenon observed for long inhibitory forms only. Importantly, KIR down-regulation on NK cells is associated with a down regulation of activating receptors (CD16, CD2 and 2B4) as well as with a lack of cell responsiveness (antibody-dependent and natural killing activities). This unresponsive state was not observed for MHC-restricted T cells. Our data implicate that, in addition to prevention of the immediate target cell lysis, KIR MHC class I interactions may also regulate the subsequent NK cell cytotoxic activity. This observation opens new perspectives in the understanding of NK cell regulation. PMID- 11385618 TI - Hapten-specific tolerance induced by acute, low-dose ultraviolet B radiation of skin requires mast cell degranulation. AB - The deleterious effects of ultraviolet B radiation (UVR) on cutaneous immunity are mediated in part by cytokines released from cutaneous cells following radiation exposure. On the one hand, TNF-alpha has been advocated as the primary mediator of failed contact hypersensitivity induction, and, on the other hand, IL 10 has been held responsible for tolerance. While keratinocytes exposed to UVR have been found to produce both TNF-alpha and IL-10, there is reason to question whether these major cellular constituents of the epidermis are the relevant source of immunomodulatory cytokines after UVR. Dermal mast cells also produce TNF-alpha and IL-10, and we have recently reported that mast cell-derived TNF alpha is required for UVR-induced impairment of CH induction. In this study, we have examined whether mast cells are also a relevant source of IL-10 in UVR dependent tolerance. We found that (a) UVR fails to induce tolerance in mast cell deficient mice, and (b) that tolerance occurs if mast cells are triggered to degranulate after ligation of the IgE receptor. Both types of tolerance were neutralized with anti-IL-10 antibodies, are hapten specific, and are associated with regulatory lymphoid cells. We conclude that mast cells are required in UVR induced tolerance and may be one of the major sources of IL-10 that mediates the tolerance induced by acute, low-dose UVR. PMID- 11385619 TI - Cytotoxic T lymphocytes recognize structurally diverse, clade-specific and cross reactive peptides in human immunodeficiency virus type-1 gag through HLA-B53. AB - Human immunodeficiency virus type-1 (HIV-1) cytotoxic T lymphocyte (CTL) epitopes have largely been defined in Caucasian populations infected with clade B virus. Identification of potentially protective CTL epitopes in non-B clade-infected African subjects is important for vaccine development. In a study of CTL responses in clade A-infected Gambians, using cytotoxicity, interferon-gamma (IFN gamma) enzyme-linked immunospot (ELISpot) and HLA-B53-peptide tetramer assays, we identified three HLA-B53-restricted epitopes in HIV-1 gag p24. CTL specific for an epitope in a highly immunogenic region of the p24 protein showed no cross reactivity to other HIV-1 clades. Two of the epitopes would not have been predicted from the peptide-binding motif due to the absence of a proline anchor at position 2. Structural analysis of HLA-B53 and its relative, HLA B35, enabled us to re-define the peptide-binding motif to include other P2 anchors. These results demonstrate the value of combined immunological and structural analyses in defining novel CTL epitopes and have implications for HIV-1 vaccine design. PMID- 11385620 TI - Thapsigargin-induced apoptosis involves Cabin1-MEF2-mediated induction of Nur77. AB - Thapsigargin (TG), which inhibits endoplasmic reticulum-dependent Ca(2 +)-ATPase and thereby increases cytosolic Ca(2 +), has been reported to cause apoptosis in T lymphocytes another cell types. In this study, we investigated the molecular mechanisms that are involved in the apoptosis induced by TG in T cell hybridomas. Exposure to TG results in rapid induction of the orphan steroid receptor, Nur77, accompanied by apoptosis of T cell hybridomas. The expression of Nur77 in response to TG treatment is sensitive to cyclosporin A, implicating that activation of calcineurin is necessary for Nur77 expression. The TG-induced Nur77 expression is also inhibited by overexpression of Cabin1, an endogenous inhibitor of calcineurin and a corepressor of the transcription factor MEF2, suggesting that MEF2 activation is required for Nur77 expression. These results suggest that induction of Nur77 expression and apoptosis by TG are mediated by the same signaling pathways that are involved in T cell receptor-mediated thymocyte apoptosis, including the calcineurin pathway and Cabin1-MEF2 pathway. PMID- 11385621 TI - More antigen-dependent CD4(+) T cell / CD4(+) T cell interactions are required for the primary generation of Th2 than of Th1 cells. AB - Mechanisms controlling the Th1 / Th2 phenotype of a primary immune response are often discussed assuming that the generation of Th1 and Th2 cells from the common CD4(+) precursor T helper (pTh) involves an interaction of this pTh cell with an antigen-presenting cell (APC) in the form of a two-cell interaction. Other studies suggest that the outcome of this two-cell interaction is modified by the presence of other T cells. No study has analyzed primary immune responses generated in normal, non-TcR transgenic mice, following the administration of a non-infectious antigen administered without adjuvant. We show that the Th1 / Th2 phenotype of such a primary response, generated in lethally irradiated recipients reconstituted with a variety of unprimed spleen cells, depends conjointly on the amount of antigen and number of unprimed syngeneic CD4(+) T cells present, with higher amounts and numbers favoring the generation of Th2 cells. Our observations show how these quantitative variables control in an interdependent manner the Th1 / Th2 phenotype of a primary immune response, and bear upon the mechanism by which this phenotype is determined. PMID- 11385622 TI - Perforin-independent regulation of dendritic cell homeostasis by CD8(+) T cells in vivo: implications for adaptive immunotherapy. AB - We investigated here the effects of perforin on CTL responses during interaction of dendritic cells (DC) with cytotoxic T lymphocytes in vivo. Using MHC class I tetramers complexed with the immunodominant CTL epitope of the lymphocytic choriomeningitis virus glycoprotein (LCMV-GP33), we followed the kinetics of DC induced CTL responses. GP33-presenting DC induced rapid primary expansion of both perforin-competent and -deficient CTL with similar kinetics. Secondary CTL responses in perforin-deficient and normal control mice after DC-booster immunization were more rapid than the primary responses, but never reached the high initial levels, suggesting that reactivated memory CTL eliminated the antigen-presenting DC and thereby limited the booster effect. Whereas killingof DC in vitro was strictly dependent on perforin, elimination of GP33-presenting DC by CTL in vivo was largely independent of perforin and Fas. Taken together, these results suggest that control of DC homeostasis by CTL, i. e. elimination of DC by the effector cells they had elicited, is controlled via multiple and probably redundant signals and represents an important fail-safe mechanism to avoid exaggerated CTL responses. PMID- 11385623 TI - Immunization with a tumor-associated CTL epitope plus a tumor-related or unrelated Th1 helper peptide elicits protective CTL immunity. AB - Immunization with cytotoxic T cell epitope SPSYVYHQF (AH1), derived from MuLV gp70 envelope protein expressed by CT26 tumor cells, does not protect BALB/c mice against challenge with CT26 tumor cells. By contrast, immunization with AH1 plus T helper peptides OVA(323-337) or SWM(106-118) eliciting Th1 and Th0 profiles, protected 83% and 33% of mice, respectively. Interestingly, immunization with AH1 plus both helper peptides reverted the efficacy to 33%. We identified the endogenous T helper peptide p(320-333) from gp70 which elicits a Th1 profile and is naturally processed. As for OVA(323-337), immunization with p(320-333) alone did not protect against tumor challenge. However, p(320-333) plus AH1 protected 89% of mice at day 10 after vaccination. Only 20% of mice vaccinated with AH1 + OVA(323-337) or AH1 + p(320-333) were protected when challenged 80 days after immunization. Treatment with OVA(323-337) or with p(320-333) around established tumors delayed tumor growth. Our results show that tumor-related as well as tumor unrelated but strong Th1 peptides may be useful for inducing CTL responses in tumor immunotherapy. PMID- 11385624 TI - The effect of HIV-1 regulatory proteins on cellular genes: derepression of the IL 2 promoter by Tat. AB - In HIV-infected individuals dysregulation of the immune system is characterized by severe disorders of the cytokine network. Increase secretion of IL-2, the major T cell growth and differentiation cytokine, may play a decisive role in sensitization of T cells for activation induced apoptosis and indirect death of activated T cells through augmented virus replication. We investigated the cause of enhanced IL-2 secretion and found that the HIV Tat induces this effect. We demonstrate that increased IL-2 secretion is due to Tat-enhanced IL-2 promoter activation. Tat derepresses and activates the distal AP-1 site (position -185 to 177) in the IL-2 promoter. In nonstimulated T cells a repressor complex containing NF-IL6, JunB, c-Fos and Fra-1 is formed on the AP-1(IL-2/d) site and represses IL-2 promoter activity. After T cell activation, a heterodimeric activator containing p65 and c-Jun binds to the AP-1(IL-2/d) site. HIV Tat enhances activation of NF-kappaB and consequently, activates the AP-1(IL-2/d) site. Our data provide evidence for a novel mechanism by which HIV Tat dysregulates IL-2 production and therefore may contribute to the HIV-1 infection in a way yet to be clarified. PMID- 11385625 TI - Two-step activation of T cells, clonal expansion and subsequent Th1 cytokine production, is essential for the development of clinical autoimmune encephalomyelitis. AB - Lewis rats immunized with guinea pig myelin basic protein (GPBP) emulsified with incomplete Freund's adjuvant (IFA) do not develop experimental autoimmune encephalomyelitis (EAE). However, we found that GPBP/IFA with pertussis toxin (PT) administration induced full-blown EAE. By comparing the immunological status of rats immunized with GPBP/IFA plus PT [PT (+) rats] with that of rats immunized with GPBP/IFA alone [PT (-) rats], we tried to elucidate the pathomechanisms of EAE. Analysis of the TCR clonality by CDR3 spectratyping revealed that Vbeta8.2 and Vbeta10 expansion of T cells occurred in both PT (-) and PT (+) rats, indicating that activation of T cells at this level is not sufficient for the development of clinical EAE. Quantitation of cytokine mRNA and protein revealed that PT (-) rats showed a Th2-dominant, while PT (+) rats showed a Th1-dominant, cytokine profile. Furthermore, administration of IL-12, but not of IFN-gamma and TNF-alpha, induced clinical EAE in GPBP/IFA-immunized animals. Taken together, two-step activation, activation of T cells bearing a particular type of TCR by antigen immunization and subsequent overproduction of Th1 cytokines, mainly IL-12 production, induced by appropriate adjuvants is essential for the development of clinical EAE. PMID- 11385626 TI - Location analysis of DNA-bound proteins at the whole-genome level: untangling transcriptional regulatory networks. AB - In this post-sequencing era, geneticists can focus on functional genomics on a much larger scale than ever before. One goal is the discovery and elucidation of the intricate genetic networks that co-ordinate transcriptional activation in different regulatory circuitries. High-throughput gene expression measurement using DNA arrays has thus become routine strategy. This approach, however, does not directly identify gene loci that belong to the same regulatory group; e.g., those that are bound by a common (set of) transcription factor(s). Working in yeast, two groups have recently published an elegant method that could circumvent this problem, by combining chromatin immunoprecipitation and DNA microarrays.(1,2) The method is likely to provide a powerful tool for the dissection of global regulatory networks in eukaryotic cells. PMID- 11385627 TI - Why girls want to be boys. AB - The mechanisms by which sex is genetically determined are bewilderingly diverse and appear to change rapidly during evolution.(1) What makes the sex-determining process so prone to perturbations? Two recent articles(2,3) explore theoretically the role of genetic conflict in sex determination evolution. Both studies use the idea that selection on sex-determining genes may act differently in parents and in offspring and they suggest that the resulting conflict can drive changes in sex-determining mechanisms. PMID- 11385628 TI - Developmental genetics and early hominid craniodental evolution. AB - Although features of the dentition figure prominently in discussions of early hominid phylogeny, remarkably little is known of the developmental basis of the variations in occlusal morphology and dental proportions that are observed among taxa. Recent experiments on tooth development in mice have identified some of the genes involved in dental patterning and the control of tooth specification. These findings provide valuable new insight into dental evolution and underscore the strong developmental links that exist among the teeth and the jaws and cranium. The latter has important implications for cladistic studies that traditionally consider features of the skull independently from the dentition. PMID- 11385629 TI - Developmental roles of platelet-derived growth factors. AB - Platelet-derived growth factor (PDGF) was originally identified in platelets and in serum as a mitogen for fibroblasts, smooth muscle cells (SMC) and glia cells in culture. PDGF has since expanded to a family of dimers of at least four gene products, whose biological actions are mediated through two receptor tyrosine kinases, PDGFRs. The present review summarizes and discusses the biological functions of PDGFs and PDGFRs in developmental processes, mainly as revealed through genetic analysis in mice. Such studies have demonstrated multiple critical roles of PDGFs and PDGFRs in embryonic and postnatal development. PDGFs seem to act upon specific populations of progenitor cells that give rise to several different cell types with distinct functions in a variety of developmental processes. Analogies are seen between the cell functions and the developmental processes controlled by PDGFs. This suggests that ancestral PDGF and PDGFR expression patterns and functions may have been iterated in related sets of morphogenetic processes in the course of evolution. PMID- 11385630 TI - Morphogenetic tissue movement and the establishment of body plan during development from blastocyst to gastrula in the mouse. AB - In many animal species, the early development of the embryo follows a stereotypic pattern of cell cleavage, lineage allocation and generation of tissue asymmetry leading to delineation of the body plan with three primary embryonic axes. The mammalian embryo has been regarded as an exception and primary body axes of the mouse embryo were thought to develop after implantation. However, recent findings have challenged this view. Asymmetry in the fertilised oocyte, as defined by the position of the second polar body and the sperm entry point, can be correlated with the orientation of the animal-vegetal and the embryonic-abembryonic axes in the preimplantation blastocyst. Studies of the pattern of morphogenetic movement of cells and genetic activity in the peri-implantation embryo suggest that the animal-vegetal axis of the blastocyst might presage the orientation of the anterior-posterior axis of the gastrula. This suggests that the asymmetry of the zygote that is established at fertilisation and early cleavage has a lasting impact on the delineation of body axes during embryogenesis. PMID- 11385631 TI - The oxidative phosphorylation (OXPHOS) system: nuclear genes and human genetic diseases. AB - The ubiquitous nature of mitochondria, the dual genetic foundation of the respiratory chain in mitochondrial and nuclear genome, and the peculiar rules of mitochondrial genetics all contribute to the extraordinary heterogeneity of clinical disorders associated with defects of oxidative phosphorylation (mitochondrial encephalomyopathies). Here, we review recent findings about nuclear gene defects in isolated OXPHOS enzyme complex deficiency. This information should help in identifying patients with mitochondrial disease and defining a biochemical and molecular basis of the disorder found in each patient. This knowledge is indispensable for accurate genetic counseling and prenatal diagnosis, and is a prerequisite for the development of rational therapies, which are still, at present, woefully inadequate. PMID- 11385632 TI - How do meiotic chromosomes meet their homologous partners?: lessons from fission yeast. AB - Homologous chromosome pairing is required for proper chromosome segregation and recombination during meiosis. The mechanism by which a pair of homologous chromosomes contact each other to establish pairing is not fully understood. When pairing occurs during meiotic prophase in the fission yeast, Schizosaccharomyces pombe, the nucleus oscillates between the cell poles and telomeres remain clustered at the leading edge of the moving nucleus. These meiosis-specific activities produce movements of telomere-bundled chromosomes. Several lines of evidence suggest that these movements facilitate homologous chromosome pairing by aligning homologous chromosomes and promoting contact between homologous regions. Since telomere clustering and nuclear or chromosome movements in meiotic prophase have been observed in a wide range of eukaryotic organisms, it is suggested that telomere-mediated chromosome movements are general activities that facilitate homologous chromosome pairing. PMID- 11385633 TI - Glycerol: a neglected variable in metabolic processes? AB - Glycerol is a small and simple molecule produced in the breakdown of glucose, proteins, pyruvate, triacylglycerols and other glycerolipid, as well as release from dietary fats. An increasing number of observations show that glycerol is probably involved in a surprising variety of physiopathologic mechanisms. Glycerol has long been known to play fundamental roles in several vital physiological processes, in prokaryotes and eukaryotes, and is an important intermediate of energy metabolism. Despite some differences in the details of their operation, many of these mechanisms have been preserved throughout evolution, demonstrating their fundamental importance. In particular, glycerol can control osmotic activity and crystal formation and then act as a cryoprotective agent. Furthermore, its properties make it useful in numerous industrial, therapeutic and diagnostic applications. Few studies have focussed directly on glycerol, however, and while its metabolism is increasingly well documented, much of the details remain unknown. Considering the importance of glycerol in multiple vital physiological processes, its study could help unlock important physiopathological mechanisms. PMID- 11385634 TI - A cellular survival switch: poly(ADP-ribosyl)ation stimulates DNA repair and silences transcription. AB - Poly(ADP-ribosyl)ation is a post-translational modification occurring in the nucleus. The most abundant and best-characterized enzyme catalyzing this reaction, poly(ADP-ribose) polymerase 1 (PARP1), participates in fundamental nuclear events. The enzyme functions as molecular "nick sensor". It binds with high affinity to DNA single-strand breaks resulting in the initiation of its catalytic activity. Activated PARP1 promotes base excision repair. In addition, PARP1 modifies several transcription factors and thereby precludes their binding to DNA. We propose that a major function of PARP1 includes the silencing of transcription preventing expression of damaged genes. Concomitant stimulation of DNA repair suggests that PARP1 acts as a switch between transcription and DNA repair. Another PARP-type enzyme, tankyrase, is involved in the regulation of telomere elongation. Tankyrase modifies a telomere-associated protein and thereby prevents it masking telomeric repeats providing access of telomerase for telomere elongation. Therefore, poly(ADP-ribosyl)ation reactions may act as molecular switches in DNA metabolism. PMID- 11385635 TI - Online analysis of development. AB - The genomics revolution has altered the very nature of research in molecular biology, from how to find genes to how to find out what specific genes do. Given the availability of so many fully (or nearly) sequenced genomes, it is now relatively easy to track down dozens or even hundreds of genes relevant to a particular field of study. Unfortunately, up till now, the tools for determining what these genes actually do in embryos and cells have not kept pace, but the burgeoning field of bioinformatics should help correct this shortcoming and introduce the power of genomics to the study of developmental biology. In this review, some of the bioinformatics resources relevant to developmental biologists are described along with some simple approaches for applying these tools to analyzing early development. PMID- 11385637 TI - A case-control study of the relationship between the risk of colon cancer in men and exposures to occupational agents. AB - BACKGROUND: We conducted a population-based case-control study in Montreal, Canada, to explore associations between hundreds of occupational circumstances and several cancer sites, including colon. METHODS: We interviewed 497 male patients with a pathologically confirmed diagnosis of colon cancer, 1514 controls with cancers at other sites, and 533 population-based controls. Detailed job histories and relevant potential confounding variables were obtained, and the job histories were translated by a team of chemists and industrial hygienists into a history of occupational exposures. RESULTS: We found that there was reasonable evidence of associations for men employed in nine industry groups (adjusted odds ranging from 1.1 to 1.6 per a 10-year increase in duration of employment), and in 12 job groups (OR varying from 1.1 to 1.7). In addition, we found evidence of increased risks by increasing level of exposures to 21 occupational agents, including polystyrene (OR for "substantial" exposure (OR(subst)) = 10.7), polyurethanes (OR(subst) = 8.4), coke dust (OR(subst) = 5.6), mineral oils (OR(subst) = 3.3), polyacrylates (OR(subst) = 2.8), cellulose nitrate (OR(subst) = 2.6), alkyds (OR(subst) = 2.5), inorganic insulation dust (OR(subst) = 2.3), plastic dusts (OR(subst) = 2.3), asbestos (OR(subst) = 2.1), mineral wool fibers (OR(subst) = 2.1), glass fibers (OR(subst) = 2.0), iron oxides (OR(subst) = 1.9), aliphatic ketones (OR(subst) = 1.9), benzene (OR(subst) = 1.9), xylene (OR(subst) = 1.9), inorganic acid solutions (OR(subst) = 1.8), waxes, polishes (OR(subst) = 1.8), mononuclear aromatic hydrocarbons (OR(subst) = 1.6), toluene (OR(subst) = 1.6), and diesel engine emissions (OR(subst) = 1.5). Not all of these effects are independent because some exposures occurred contemporaneously with others or because they referred to a group of substances. CONCLUSIONS: We have uncovered a number of occupational associations with colon cancer. For most of these agents, there are no published data to support or refute our observations. As there are few accepted risk factors for colon cancer, we suggest that new occupational and toxicologic studies be undertaken focusing on the more prevalent substances reported herein. PMID- 11385638 TI - Nested case-control study of lung cancer among pulp and paper workers in relation to exposure to dusts. AB - BACKGROUND: Numerous studies have indicated an increased risk of lung cancer in pulp and paper industry workers. In a 1990 survey, standardized mortality ratio (SMR) was found to be 122 (95% CI:96-153) for lung cancer in Polish male workers in the pulp and paper industry, and 166 (95% CI:95-270) among workers engaged in paper production. METHODS: A nested case-control design within a cohort of pulp and paper workers was applied. Seventy-nine lung cancer cases and 237 "healthy" controls were selected from the cohort of 10,460 workers employed during the years 1968-1990, and observed until the end of 1995. Based on personnel files, occupational exposure was reconstructed by experts. Using a questionnaire, data on smoking habits were collected. ORs unadjusted and adjusted for smoking were calculated applying the model of conditional logistic regression. RESULTS: Occupational exposure to inorganic dusts (kaolin, lime, cement, brick, grindstone) adjusted for smoking was a significant lung cancer risk factor, with a 4.0-fold risk (95% CI:1.3-12.6), and a dose-response by cumulative dose index. Among organic dusts only wood dust increased albeit insignificantly the risk for those exposed (adjusted for smoking OR = 2.1, 95% CI:0.9-4.9), but without dose response relationship. CONCLUSIONS: Exposure to occupational dust with relatively low content of silica, but at high concentrations may be considered as a factor increasing lung cancer risk. However, the observation made in this study should be viewed with caution as it was based on a small number of cases, and further evidence is needed to confirm or refute the authors' hypothesis. PMID- 11385639 TI - Nested case-control analysis of high pesticide exposure events from the Agricultural Health Study. AB - BACKGROUND: A nested case-control analysis of high pesticide exposure events (HPEEs) was conducted using the Iowa farmers enrolled in the Agricultural Health Study (AHS). METHODS: In the 12 months of the study, 36 of the 5,970 farmer applicators randomly chosen from the AHS cohort (six per 1,000 farmer applicators per year) met our definition of an HPEE, by reporting "an incident with fertilizers, weed killers, or other pesticides that caused an unusually high personal exposure" resulting in physical symptoms or a visit to a health care provider or hospital. Eligibility criteria were met by 25 HPEE cases and 603 randomly selected controls. RESULTS: Significant risk factors for an HPEE included: poor financial condition of the farm which limited the purchase of rollover protective structures OR = 4.6 (1.5-16.6), and having a high score on a risk acceptance scale OR = 3.8 (1.4-11.2). Other non-significant factors were also identified. CONCLUSIONS: The limited statistical power of this study necessitates replication of these analyses with a larger sample. Nonetheless, the observed elevated odds ratios of an HPEE provide hypotheses for future studies that may lead to preventive action. PMID- 11385640 TI - Occupational categories at risk for Parkinson's disease. AB - BACKGROUND: The etiology of Parkinson's disease (PD) is considered to have a strong environmental component, but relatively few studies have investigated the potential association between occupation and the disease. METHODS: In a population-based case-control study, we collected comprehensive occupational histories from all study participants, 144 case and 464 control subjects. RESULTS: Chi-square analysis revealed that working in an agricultural occupation increased estimated PD risk (OR = 1.74; 95% CI = 0.85, 3.60). In contrast, a history of ever working in a service occupation was negatively associated with PD risk (OR = 0.69; 95% CI = 0.47, 1.00). Risk estimates were close to one for specific service occupations. Adjusted odds ratios for all non-service occupational and industrial categories were similar, and working in a service occupation was the only significant inverse predictor of PD risk. CONCLUSIONS: Future investigations focusing on lifestyle factors and environmental exposures within the agricultural and service occupational categories are warranted. PMID- 11385641 TI - Occupational exposures and cancers of the endometrium and cervix uteri in Finland. AB - BACKGROUND: Endometrial cancer incidence rates are low in Asia and Africa and high in North America and Northern Europe. Cervical cancer is often the most common female cancer in developing countries, and infection with human papillomavirus (HPV) is its main risk factor. However, other factors, such as occupational exposures may modify the HPV-related risk. We conducted an exploratory register-linkage study in Finland to assess the role of occupational exposures on incidence rates of cancers of the endometrium and cervix uteri. METHODS: Occupational risk factors for endometrial and cervical cancers were explored in a 25-year follow-up of female workers born 1906-1945 (N = 413,877) identified through the Population Census of Finland of 1970. Job titles in census records were converted to exposures of 31 occupational agents through a job exposure matrix. Poisson regression models estimated relative risks (RR) for each agent, standardized for birth cohort, follow-up period, and socio-economic status. For each agent, the product of level and probability of exposure was calculated and subdivided in three categories: zero, low, and medium/high. Adjustment at the job title level was done for the turnover rate (endometrial and cervical cancers), mean parity, and age at first birth (endometrial cancer). RESULTS: Endometrial cancer (2,833 cases) was associated with exposure to animal dust (RR 1.2, low level, 174 cases) and sedentary work (RR 1.3, high level, 145 cases). Cervical cancer (1,101 cases) was associated with exposure to aliphatic and alicyclic (RR 1.3, low level, 91 cases), aromatic (RR 1.2, low level, 318 cases; RR 1.4, high level, 41 cases), and chlorinated hydrocarbon solvents (RR 1.3, low level, 50 cases), silica dust (RR 1.2, low level, 251 cases), and wood dust (RR 1.2, low level, 249 cases). CONCLUSIONS: This study suggests that occupational exposures may be associated with increased risk of endometrial and cervical cancers. PMID- 11385642 TI - Epidemiology of occupational injuries and illnesses in a university population: a focus on age and gender differences. AB - BACKGROUND: Occupational injuries and illnesses are a major preventable public health problem. METHODS: This study evaluated the incidence, nature, and cause of awarded workers' compensation claims for a large state university and teaching hospital. Rates and types of injury were compared across age and gender. RESULTS: Rates of injury varied over twofold with age, with those 16-25 having the lowest rates of injury and those 36-45 having the highest rates. Claims rates were 1.36 fold higher for women than men. Women had higher rates for injury resulting from lifting, falling, noxious exposures, repetitive motion, and carpal tunnel syndrome. Similarly women had significantly higher rates of claims for pain, sprains, bruises, burns, concussion, and inhalation injury; with lower rates of cuts, ligament injury, and jammed joints. CONCLUSIONS: These group differences suggest the need to examine age and gender job distributions and relevant ergonomic and environmental causative factors. PMID- 11385643 TI - Subclinical immunologic and physiologic responses in hexamethylene diisocyanate exposed auto body shop workers. AB - BACKGROUND: Diisocyanates are potent sensitizing agents and currently the most commonly identified cause of occupational asthma in industrialized countries. However, diisocyanate asthma is difficult to diagnose and exposure and host risk factors are unclear. Auto body shops, one of the most common hexamethylene diisocyanate (HDI) exposure settings, are particularly difficult to study due to their small size and episodic exposures. Surveillance studies of such workers are limited. OBJECTIVES: We have initiated a cross-sectional field epidemiologic study, Survey of Painters and Repairers of Auto bodies by Yale (SPRAY), to characterize the effects of diisocyanate exposures on actively employed auto body shop workers. Methods and Results We present here questionnaire, physiologic, immunologic, and exposure data on 75 subjects enrolled in the study. No overt cases of clinically apparent diisocyanate asthma were identified based on spirometry, methacholine challenge, peak flows, and symptoms. HDI-specific lymphocyte proliferation was present in 30% of HDI-exposed workers and HDI specific IgG in 34% of HDI-exposed workers, but they were not associated. HDI specific IgE was detected in two workers. HDI-specific lymphocyte proliferation, increased methacholine responsiveness, and symptoms of chest tightness and shortness of breath were more common in the most heavily HDI-exposed workers, the painters. More long-term follow-up of this cohort should clarify the significance of these HDI-specific immunologic responses, physiologic changes, and symptoms. CONCLUSIONS: These findings demonstrate the presence of HDI-specific immune responses in a large proportion of healthy HDI-exposed workers. PMID- 11385644 TI - Respiratory symptoms among crab processing workers in Alaska: epidemiological and environmental assessment. AB - BACKGROUND: Crab processing workers may develop respiratory symptoms and specific IgE responses, but the risk factors have not been fully described. METHODS: In 1998, 107 workers at a crab processing facility completed a survey both at the beginning and end of the processing season. The surveys included standardized symptom questionnaires, spirometry, and serological testing, as well as measurement of workplace airborne crab allergens and microscopic analysis of aerosolized materials. RESULTS: Over the crab processing season, asthma-like symptoms developed in 26% of study participants and bronchitic symptoms in 19%. Only 9% of those with new asthma-like symptoms were IgE-sensitized to crab at the end of the season. Among the crab processing jobs, butchering and degilling workers had the highest incidence of respiratory symptoms. CONCLUSIONS: Both personal and process-related factors appear to affect the development of respiratory symptoms in crab processing workers. In this study, crab specific IgE was not detected in most of the workers with new symptoms. Published 2001 Wiley Liss, Inc. PMID- 11385645 TI - Asthma and other respiratory symptoms in New Zealand pine processing sawmill workers. AB - BACKGROUND: To study respiratory symptoms in pine sawmill workers. METHODS: A respiratory health questionnaire was administered to 772 pine sawmill workers and the association between symptoms and job-title-based exposure was studied. RESULTS: Asthma in exposed workers (18%, n = 704) was more common than in the general population (12.1%, n = 592; adjusted OR (95% CI): 1.6 (1.1-2.3)). Asthma was also more common in the low exposure group (15.6%, n = 294) and high exposure groups (high exposure to 'green dust'; 20.4%, n = 212 and high exposure to 'dry dust'; 18.8%, n = 198) than in the non-exposed workers (9.2%, n = 65). Adjusted odds ratios were 1.9 (0.7-4.9), 2.7 (0.9-7.6), and 2.1 (0.8-5.7), respectively. Adjusted odds ratios for symptoms of cough were 2.7 (1.2-6.5) for the low, 5.2 (2.1-13.0) for the high 'green dust' and 3.3 (1.4-7.9) for the high 'dry dust' exposure groups. Moreover, eye and nose irritations were significantly more prevalent in the high and low exposure groups. CONCLUSIONS: Working in pine sawmilling is associated with an increased prevalence of asthma and cough symptoms and eye and nose irritation. PMID- 11385646 TI - Hypersensitivity pneumonitis in a metal-working environment. AB - BACKGROUND: An outbreak of lung disease among workers in a metal-working plant included 16 biopsy-confirmed cases of hypersensitivity pneumonitis and additional patients with asthma, bronchiolitis and emphysema, usual interstitial pneumonitis, and sarcoidosis. Study design Clinical examination of patients; cross-sectional questionnaire survey of the outbreak plant and two control plant areas, one with and one without MWF exposures, in a separate facility; industrial hygiene survey with laboratory characterization of microbial flora; and immunological investigation. METHODS: Patients with suspected hypersensitivity pneumonitis underwent a clinical examination including detailed lung function, imaging, and tissue studies. A plant walk-through identified metal-working processes, microbial aerosols, and work practices. Microbial characteristics of the three microbial aerosol-producing processes were characterized. Antibodies to those agents were determined in patient sera. A questionnaire survey was conducted in the case plant and in two areas of a control plant, one with and one without metal-working fluids exposure. RESULTS: Thirty-nine (79.6%) patients described symptoms consistent with work-related lung disease, eight received other diagnoses, and two did not complete their examinations. Sixteen patients had hypersensitivity pneumonitis confirmed on biopsy. Mean decrements in lung forced expiratory volume in 1 s and force vital capacity from before to after work were similar in the 16 biopsy-confirmed cases of hypersensitivity pneumonitis ( - 6.3%; - 7.2%) and the 19 symptomatic patients without biopsies ( 11.2%, - 10.1%). Symptoms were more common in the case plant than in a non-MWF control plant area. Three sources of water-based aerosols were identified that grew similar microbial flora. Although machining increased airborne bacterial levels, the increase was not related to the concentration of viable bacteria in the sumps. Antibody testing did not identify a specific single organisms. Endotoxin levels were similar in case and MWF control plant. CONCLUSIONS: Lung disease in environments with water-based aerosols may be more common than usually recognized. Patients with HP often present with only subtle abnormalities and may be missed if multiple clinical abnormalities are required to document disease. PMID- 11385647 TI - What is the most cost-effective way to identify silica problem worksites? AB - BACKGROUND: State based surveillance systems to identify cases of silicosis have been developed over the past 18 years to target worksite interventions to reduce the incidence of silicosis. Using data from the Michigan silicosis surveillance system, we conducted an analysis to determine the most cost-effective way to identify problem worksites. METHODS: The initial reporting source of all 470 confirmed cases of silicosis reported to the Michigan surveillance system from 1989 to 1995 was identified. The cost of identifying confirmed cases, worksites, problem worksites, silica problem worksites, and the number of current silica exposed workers was determined for four reporting sources: hospitals; physicians; workers' compensations; and death certificates. RESULTS: Hospital reports were the first to identify 67% of the confirmed cases, 74% of the worksites, and 58% of the problem worksites. Physician reports initially identified 17% of confirmed cases, 15% of worksites, and 26% of problem worksites. Workers' compensation records initially identified 11% of confirmed cases, 4% of worksites, and 8% of problem worksites. Death certificates initially identified 5% of confirmed cases, 7% of worksites, and 8% of problem worksites. Hospital reports were the most cost effective way to identify cases (US$ 143), worksites (US$ 313), and problem worksites (US$ 454). CONCLUSIONS: Hospital discharge records identified the greatest number of confirmed cases and problem worksites and was the most cost effective approach to identify both individuals with silicosis and worksites with problems. PMID- 11385648 TI - Capture-recapture estimation of unreported work-related musculoskeletal disorders in Connecticut. AB - BACKGROUND: Estimates of the extent of musculoskeletal disorders (MSD) are usually based upon workers' compensation reports, although recent reports indicate that there may be widespread under-reporting of MSD. METHODS: An estimate of the incidence of arm and hand work-related MSD was made using capture recapture analysis of the overlap between state workers' compensation reports and physician reports in Connecticut for 1995. The resulting estimate was compared to a population-based survey of MSD. RESULTS: There was very small overlap between the two state injury reporting systems: 6.7% of 793 reported workers' compensation cases, or 8% of 661 physician's reports. The estimate for MSD not captured by either system was 13,285, resulting in 14,686 (95% CI: 9,733-18,453) total reported and non-reported cases. This compares to an estimate of 13,775 cases (95% CI: 8,800-18,800) based on a phone survey. CONCLUSIONS: This analysis points to substantial under-reporting of MSD in Connecticut: estimates of unreported cases exceed those officially reported by a factor of 11:1. The findings have an important bearing on injury prevention programs and policy making. PMID- 11385651 TI - Water: From Clusters to the Bulk. AB - Water is of fundamental importance for human life and plays an important role in many biological and chemical systems. Although water is the most abundant compound on earth, it is definitely not a simple liquid. It possesses strongly polar hydrogen bonds which are responsible for a striking set of anomalous physical and chemical properties. For more than a century the combined importance and peculiarity of water inspired scientists to construct conceptual models, which in themselves reproduce the observed behavior of the liquid. The exploration of structural and binding properties of small water complexes provides a key for understanding bulk water in its liquid and solid phase and for understanding solvation phenomena. Modern ab initio quantum chemistry methods and high-resolution spectroscopy methods have been extremely successful in describing such structures. Cluster models for liquid water try to mimic the transition from these clusters to bulk water. The important question is: What cluster properties are required to describe liquid-phase behavior? PMID- 11385649 TI - Exposure to endosulfan in farmers: two case studies. AB - BACKGROUND: Endosulfan is not a restricted use organochlorine insecticide and is currently under re-registration review. In 1993, one confirmed case and one possible case of endosulfan poisoning in agricultural workers occurred in two southeastern states. METHODS: Two cases of suspected endosulfan poisoning were investigated utilizing record reviews, blood samples, a site visit, and clothing analysis. RESULTS: Case 1 was fatal; Case 2 resulted in permanent neurological impairment. Additionally, Case 1 mixed and applied two less toxic pesticides, acephate and maleic hydrazide to tobacco plants. Both farm owners had ample opportunity for endosulfan exposure while mixing concentrated endosulfan with water and applying the solution to tobacco with boom sprayers pulled by tractors. CONCLUSIONS: Estimates of the absorbed dose of endosulfan were not available because methods to determine actual personal exposure that would be found in fat or tissue samples were not used. Health and safety issues associated with endosulfan require a closer examination. A cooperative multi-disciplinary approach to providing timely accurate education is needed to prevent pesticide poisonings. PMID- 11385652 TI - Transition Metal Catalysis Using Functionalized Dendrimers. AB - Dendrimers are well-defined hyperbranched macromolecules with characteristic globular structures for the larger systems. These novel polymers have inspired many chemists to develop new materials and several applications have been explored, catalysis being one of them. The recent impressive strides in synthetic procedures increased the accessibility of functionalized dendrimers, resulting in a rapid development of dendrimer chemistry. The position of the catalytic site(s) as well as the spatial separation of the catalysts appears to be of crucial importance. Dendrimers that are functionalized with transition metals in the core potentially can mimic the properties of enzymes, their efficient natural counterparts, whereas the surface-functionalized systems have been proposed to fill the gap between homogeneous and heterogeneous catalysis. This might yield superior catalysts with novel properties, that is, special reactivity or stability. Both the core and periphery strategies lead to catalysts that are sufficiently larger than most substrates and products, thus separation by modern membrane separation techniques can be applied. These novel homogeneous catalysts can be used in continuous membrane reactors, which will have major advantages particularly for reactions that benefit from low substrate concentrations or suffer from side reactions of the product. Here we review the recent progress and breakthroughs made with these promising novel transition metal functionalized dendrimers that are used as catalysts, and we will discuss the architectural concepts that have been applied. PMID- 11385653 TI - Blue Fluorescent Exciplexes Consisting of trans-Stilbene and Antibodies. PMID- 11385654 TI - Retracing Enzyme Evolution in the (betaalpha)(8)-Barrel Scaffold. PMID- 11385655 TI - Cyclic Dimers of Metalloporphyrins as Tunable Hosts for Fullerenes: A Remarkable Effect of Rhodium(III) We thank Dr. F. Hasegawa of the Science University of Tokyo for HR-MS measurements. J.-Y.Z. and K.S. thank the JSPS for financial support. PMID- 11385656 TI - Photoelectrochemistry with Controlled DNA-Cross-Linked CdS Nanoparticle Arrays This research is supported by The U.S.-Israel Binational Science Foundation. The Max Planck Research Award for International Cooperation (I.W.) is gratefully acknowledged. PMID- 11385657 TI - The Main Group Macrocycle PMID- 11385658 TI - Experimental and Theoretical Observations of Aromaticity in Heterocyclic XAl(3)( ) (X=Si, Ge, Sn, Pb) Systems The experimental work reported herein was supported by the National Science Foundation (DMR-0095828) and was performed at the W. R. Wiley Environmental Molecular Sciences Laboratory, a national scientific user facility sponsored by the DOE's Office of Biological and Environmental Research and located at the Pacific Northwest National Laboratory, which is operated for DOE by the Battelle Memorial Institute. L.S.W. is an Alfred P. Sloan Foundation Research Fellow. The theoretical work was carried out at the Utah State University and was supported by the donors of The Petroleum Research Fund (ACS PRF 35255-AC6), administered by the American Chemical Society. PMID- 11385659 TI - Template-Directed Synthesis of a PMID- 11385660 TI - Precipitons-Functional Protecting Groups to Facilitate Product Separation: Applications in Isoxazoline Synthesis. PMID- 11385661 TI - Probing Guest Geometry and Dynamics through Host-Guest Interactions This work was supported by the CREST (Core Research for Evolutional Science and Technology) project of Japan Science and Technology Corporation. PMID- 11385662 TI - Chiral Salen-Aluminum Complexes as Catalysts for Enantioselective Aldol Reactions of Aldehydes and 5-Alkoxyoxazoles: An Efficient Approach to the Asymmetric Synthesis of syn and anti beta-Hydroxy-alpha-amino Acid Derivatives Financial support has been provided by the National Institutes of Health (GM 43912-11 and GM 33328-18). PMID- 11385663 TI - Fluorescent Fingerprinting of Molecular Recognition Landscapes J.J.L. thanks Wolfgang Rettig (Humboldt University) for his generosity during the preparation of this manuscript and the Uebbing Foundation for support. M.A. thanks Jan E. de Vries for continuous support and encouragement. PMID- 11385664 TI - Direct Observation of Surface-Controlled Self-Assembly of Coordination Cages by Using AFM as a Molecular Ruler We acknowledge the Nanolink Program of the MESA(+) Research Institute (University of Twente), the CNR Nanotechnology Programme, and MURST (Project Molecular Nanoelectronics) for financial support of this work. A special thanks goes to Dr. Maik Liebau (University of Twente) for the preparation of the microcontact-printed substrates and Dr. Frank Geurts (AKZO NOBEL, Central Research Arnhem, NL) for the XPS measurements. PMID- 11385665 TI - Highly Enantioselective Allylation of Imines with a Chiral Zirconium Catalyst This work was partially supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture, Japan. The authors are grateful to Dr. Motoo Shiro (Rigaku Co. Ltd.) for his help in performng the X-ray analysis of compound 3 cd. T.G. thanks the Japan Society for the Promotion of Science (JSPS) for the award of a postdoctoral research fellowship. PMID- 11385666 TI - Noncoordinating Dendrimer Polyanions: Cocatalysts for the Metallocene-Catalyzed Olefin Polymerization. PMID- 11385667 TI - The First Barium-Carborate Complex: Synthesis and Structural Investigation This work was supported by the National Science Foundation (CHE-9988045) and the Petroleum Research Fund, administered by the American Chemical Society. C.G. thanks the state of Bavaria, Germany for a scholarship. PMID- 11385668 TI - NSCl(2)(-): Thiazyl Dichloride-An Aza Analogue of Thionyl Dichloride A.S. thanks Prof. Dr. T. M. Klapotke (LMU Munchen) for his generous support and Prof. Dr. R. D. Harcourt (Melbourne University) for many helpful discussions. We thank Prof. Dr. W. Schnick and S. Schmidt (LMU Munchen) for carrying out the powder diffraction experiments, Dr. K. Karaghiosoff and Dr. J. Senker (MAS) for carrying out the NMR experiments. PMID- 11385669 TI - A Homoleptic Carbene--Lithium Complex Homoleptic carbene complexes part 10. This work was supported by the Fonds der Chemischen Industrie, the Deutsche Forschungsgemeinschaft, and by the Studienstiftung des Deutschen Volkes. Part 9 ref. 14. PMID- 11385670 TI - Domino Hydroformylation/Knoevenagel/Hydrogenation Reactions This work was supported by the Fonds der Chemischen Industrie (Fellowship to S.K.Z.) and the Deutsche Forschungsgemeinschaft, as well as an Alfried Krupp Award for Young University Teachers from the Krupp foundation. We thank the companies BASF and Degussa for their gift of chemicals. PMID- 11385671 TI - Dialkenylation of Carbonyl Groups by Alkenyllithium Compounds: Formation of Cyclopentadiene Derivatives by the Reaction of 1,4-Dilithio-1,3-dienes with Ketones and Aldehydes This work was partially supported by the National Science Fund for Distinguished Young Scholars (29825105), the Major State Basic Research Development Program (G2000077502-D), and the National Natural Science Foundation of China. The Qiu Shi Science & Technologies Foundation is gratefully acknowledged. PMID- 11385672 TI - [Me] PMID- 11385673 TI - Hydrothermal Synthesis, Structure, and Magnetism of PMID- 11385674 TI - A Convenient Method for the Generation of a Disulfur Monoxide Equivalent and Its Reaction with Diazoalkanes to Yield Dithiirane 1-Oxides This work was supported by a Grant-in-Aid for Scientific Research (No. 90193242) from the Ministry of Education, Science, Sports, and Culture, Japan. PMID- 11385675 TI - Solid-State Isomerization of Atropodiastereomers: Effective Diastereoselection through Polymorphic Transformations Prof. Jean-Louis Pierre and Prof. Jacques Reisse are gratefully acknowledged for fruitful discussions, as well as Dr. Christian Philouze for assistance with the X-ray crystallographic studies. We would also like to thank one of the referees for valuable suggestions. We thank Universite Joseph Fourier and CNRS for financial support. PMID- 11385676 TI - Rhodium-Catalyzed Hydrophosphorylation of Terminal Alkynes Leading to Highly Selective Formation of (E)-Alkenylphosphonates: Complete Reversal of Regioselectivity to the Palladium-Catalyzed Counterpart We are grateful to the Japan Science and Technology Corporation (JST) for financial support through the CREST (Core Research for Evolutional Science and Technology) program and for a postdoctoral fellowship to C.-Q.Z. PMID- 11385677 TI - Mechanistic Rationale of a Palladium-Catalyzed Allylic Substitution Polymerization-Carbon-Carbon Bond-Forming Polycondensation out of Stoichiometric Control by Cascade Bidirectional Allylation This work was supported by a Grant-in Aid for Scientific Research (No. 12750778) from the Ministry of Education, Science, Sports, and Culture, Japan. A fellowship to K.T. from the Japan Society for the Promotion of Science for Young Scientists is gratefully acknowledged. In this contribution, the term "out of stoichiometric control" means that when reagent 2 a is used in excess, the polymerization is under stoichiometric control, whereas when 1 is in excess, the polymerization is not under stoichiometric control. PMID- 11385678 TI - Kinetic Reactivity of "Higher Order Cuprates" in S(N)2 Alkylation Reactions We thank Prof. A. Alexakis and Prof. B. Breit for providing unpublished results.11 Generous allotment of computer time from the Institute of Molecular Science and the Intelligent Modeling Laboratories, the University of Tokyo, is gratefully acknowledged. PMID- 11385679 TI - Ionization of O(3) in Excess N(2): A New Route to N(2)O via Intermediate N(2)O(3)(+) Complexes This work was supported by the University of Rome "La Sapienza", the University of Perugia, the Italian National Research Council (CNR), and the Ministero dell'Universita e della Ricerca Scientifica e Tecnologica (MURST). The authors are grateful for the skilled help of F. Angelelli and A. Di Marzio. PMID- 11385680 TI - Preorganization of the Bioactive Conformation of Sialyl Lewis(X) Analogues Correlates with Their Affinity to E-Selectin. PMID- 11385681 TI - Benzene-Free Synthesis of Phenol This research was supported by a grant awarded by the U.S. Department of Agriculture. PMID- 11385682 TI - Highly Efficient Catalytic Synthesis of alpha-Amino Acids under Phase-Transfer Conditions with a Novel Catalyst/Substrate Pair This work was supported by an ISTC (grant A-356), by the Russian Foundation for Basic Research (Project No 99 03-32970), the Grant Agency of the Czech Republic (Grant No. 203/00/0601), and also by the Grant Agency of Charles University (Grant No. 18/98). We thank Prof. Pavel Kocovsky for encouragement and proofreading the manuscript. PMID- 11385683 TI - Synthesis and Characterization of a Metallabenzyne This work was supported by the Hong Kong Research Grant Council and The Hong Kong University of Science and Technology. We thank Professor C. P. Lau for valuable discussions. PMID- 11385684 TI - Characterization of the Alkylation Product of Heme by the Antimalarial Drug Artemisinin We are grateful to the CNRS for financial support, and to the French Ministery of Education for a PhD grant to J.C. Dr. Yannick Coppel (LCC-CNRS) is gratefully acknowledged for discussions on NMR data. PMID- 11385685 TI - Assembly of Ni(7) and Ni(21) Molecular Clusters by Using Citric Acid This work was supported by the Swiss National Science Foundation. PMID- 11385686 TI - Synthesis of a Cyclopropene Analogue of Ceramide, a Potent Inhibitor of Dihydroceramide Desaturase This work was supported by the Direccion General de Ensenanza Superior e Investigacion Cientifica (grant PB97-1171) and the Departament d'Universitats, Recerca i Societat de la Informacio, Generalitat de Catalunya (grant 1999-SGR 00187 and a Predoctoral fellowship to G.T.). We thank Dr. J. Casas, Dr. A. Delgado, and Dr. J. Joglar for their help in different aspects of this work. PMID- 11385687 TI - Synthesis of Cored Dendrimers with Internal Cross-Links This work was supported by the National Institute of Health (GM 39782). L.G.S. acknowledges the Department of Chemistry, University of Illinois, and Pharmacia & Upjohn for fellowship support. PMID- 11385688 TI - Synthesis of the Functionalized Macrocyclic Core of Proteasome Inhibitors TMC-95A and B This work was supported by grants from the National Institutes of Health (grant CA28824). We thank Dr. George Sukenick and Sylvi Rusli of the MSKCC NMR Core Facility for NMR and mass spectral analyses (NIH Grant CA08748). PMID- 11385689 TI - Life, liberty, the pursuit of happiness,... and health care? PMID- 11385690 TI - BPH with coexisting overactive bladder dysfunction--an everyday urological dilemma. AB - The aim of this study was to use a systematic schedule, including urodynamics, to describe the rate of coexisting overactive bladder (OB) in patients with bladder outlet obstruction (BOO) caused by benign prostatic hyperplasia (BPH). We also identified differences between the patients with pure BOO compared with those with BOO combined with OB (BOO + OB). One hundred and sixty-two men referred to our clinic due to LUTS were included. Patients with a history that might affect their bladder function were excluded. After cystometry and pressure-flow studies, the patients were divided into pure BOO and BOO + OB. Of the 162 men, 55% had pure BOO. BOO + OB was found in 45%. Age, s-PSA, voided volume, and obstruction grade differed significantly between the groups. The patients with BOO + OB were older, had a higher s-PSA, voided smaller volumes, and were more obstructed. We found no differences in TRUS-volume, Q-max, IPS score, or PVR. There was a strong association between OB and BOO, the percentage of OB increasing with increased obstruction. TRUS-volume, Q-max, IPS score, and PVR did not predict whether the patients had a combined BOO + OB or not. These findings indicate that BOO is a progressive disease, which in time causes pronounced obstruction and perhaps in itself contributes to the development of OB. PMID- 11385691 TI - Involuntary detrusor contractions: correlation of urodynamic data to clinical categories. AB - Data regarding the prevalence and urodynamic characteristics of involuntary detrusor contractions (IDC) in various clinical settings, as well as in neurologically intact vs. neurologically impaired patients, are scarce. The aim of our study was to evaluate whether the urodynamic characteristics of IDC differ in various clinical categories. One hundred eleven consecutive neurologically intact patients and 21 consecutive neurologically impaired patients, referred for evaluation of persistent irritative voiding symptoms, were prospectively enrolled. All patients were presumed by history to have IDC, and underwent detailed clinical and urodynamic evaluation. Based on clinical evaluation, patients were placed into one of four categories according to the main presenting symptoms and the existence of neurological insult: 1) frequency/urgency; 2) urge incontinence; 3) mixed stress incontinence and irritative symptoms; and 4) neurogenic bladder. IDC was defined by detrusor pressure of > or = 15 cm H2O whether or not the patient perceived the contraction; or < 15 cm H2O if perceived by the patient. Eight urodynamic characteristics of IDC were analyzed and compared between the four groups. IDC were observed in all of the neurologically impaired patients, compared with 76% of the neurologically intact patients (P < 0.001). No correlation was found between amplitude of IDC and subjective report of urgency. All clinical categories demonstrated IDC at approximately 80% of cystometric capacity. Eighty-one percent of the neurologically impaired patients, compared with 97% of the neurologically intact patients, were aware of the IDC at the time of urodynamics (P < 0.04). The ability to abort the IDC was significantly higher among continent patients with frequency/urgency (77%) compared with urge incontinent patients (46%) and neurologically impaired patients (38%). In conclusion, when evaluating detrusor overactivity, the characteristics of the IDC are not distinct enough to aid in differential diagnosis. However, the ability to abort IDC and stop incontinent flow may have prognostic implications, especially for the response to behavior modification, biofeedback, and pelvic floor exercise. PMID- 11385692 TI - Learned voiding dysfunction (non-neurogenic, neurogenic bladder) among adults. AB - Data concerning learned voiding dysfunction (Hinman syndrome; non-neurogenic, neurogenic bladder) in adults are scarce. The present study was conducted to assess the pre-valence and clinical characteristics of this dysfunction among adults referred for evaluation of lower urinary tract symptoms. Learned voiding dysfunction was suggested by a characteristic clinical history and intermittent "free" uroflow pattern and by the absence of any detectable neurological abnormality or anatomic urethral obstruction. A definitive diagnosis was made by the demonstration of typical external urethral sphincter contractions during micturition by EMG or fluoroscopy. A urodynamic database of 1,015 consecutive adults was reviewed. Twenty-one (2%) patients (age, 24-76 years) met our strict criteria of learned voiding dysfunction. Obstructive symptoms were the most common presenting symptoms, followed by frequency, nocturia, and urgency. Eight (35%) patients had recurrent urinary tract infections, seven of these being women. None of the patients had any clinically significant upper urinary tract damage. First sensation volume was significantly lower in women than in men. Both detrusor pressure at maximum flow and maximum detrusor pressure during voiding were found to be significantly higher in men than in women. Further differentiation between adult women and men failed to reveal any other clinically significant differences. In conclusion, by strict video-urodynamic criteria, 2% of our patients had learned voiding dysfunction. Other patients, with presumed learned voiding dysfunction, who did not undergo video-urodynamics were not included in the present series. Thus, the prevalence of learned voiding dysfunction among adults referred for evaluation of lower urinary tract symptoms is likely to be even higher. PMID- 11385693 TI - Pelvic muscle activity in nulliparous volunteers. AB - The aim of this study was to describe normal characteristics of spontaneous and voluntary pelvic muscle function in nulliparous healthy continent women and to assess the reaction of the pelvic floor to stress and fatigue. Ten nulliparous volunteers were recruited. Pelvic muscle strength was evaluated by palpation and perineal ultrasound. Kinesiological EMG and perineal ultrasound were performed to test for possible fatigue and to assess bladder neck mobility during coughing with a pre-contraction of the pelvic floor muscles. Bladder neck mobility did not increase after attempts to fatigue the pelvic floor muscles. Bladder neck descent was significantly less when the women were instructed to contract the pelvic floor muscles before coughing. The contraction of the pelvic floor muscles stabilizes the vesical neck in nulliparous women. PMID- 11385694 TI - An ultra-short perineal pad-test for evaluation of female stress urinary incontinence treatment. AB - We assessed the reproducibility and feasibility of a rapid perineal pad-test designed for evaluation of stress urinary incontinence treatment. In an experimental study, we included 34 women with genuine stress incontinence, 13 with urge incontinence and 10 non-incontinent volunteers for a repeated pad-test with a standardized bladder volume (300 mL) and a standardized physical activity during one minute. The pad was weighed before and after the exercise to estimate the volume of any leaked urine. In order to find out the percentage of unselected, incontinent women who had been able to perform a preoperative pad test, we identified all 120 women operated on for stress incontinence during a one-year period in a separate retrospective analysis. In another retrospective analysis, we compared the subjective outcome of laparoscopic colposuspension with the outcome of the test performed before and after the procedure among 93 women with genuine stress incontinence. Among stress incontinent women, the median of the differences in leakage between the first and the second test was 8.5 mL (range 0-60 mL) and the repeatability coefficient was 33.6 mL. None of the 13 women with urge incontinence leaked during the tests. One of the 10 controls leaked (during both tests). Of the unselected women, 104/120 (87%) were able to perform the test. Reduction of leaking after surgery was significantly associated with subjective report of outcome (P < 0.0001). In conclusion, the described pad test has an acceptable reproducibility and feasibility making it suitable for follow-up studies. PMID- 11385695 TI - Influence of gender on the diurnal variation of urine production and micturition characteristics of the rat. AB - The diurnal variation in the frequency/volume characteristics of male and female conscious rats was evaluated with reference to fluid consumption and urine production. Baseline values of the micturition volume and frequency of nine male and 10 female SD adult rats were measured over a 24-hour time period. The level of initial hydration conditions was standardized with 5 ml of water administered orally. With animals in a metabolism chamber having free access to water, the total volume of water consumed, the frequency/volume characteristics during micturition and the urine production rate were derived from the measurements of voided volume as detected by a digital balance. To establish reliability of measurements two separate micturition studies were done per rat at an interval of 1 week. Mean frequency of micturition and mean volume voided per micturition and urine production rate were computed in 3-hour time bins and represented over the 24-hour period. In addition the mean values of the number of micturitions and mean micturated volumes during the day/dark cycle were evaluated. The results show significant gender specificity in water consumption, urine production, and diurnal variations in micturition frequency/volume characteristics. Females consistently consume significantly larger amounts of water (83%) than males while urine production rate was correspondingly higher in females. It is concluded that water consumption and urine production are gender-specific. Because higher volumes of water are imbibed by females than males, the frequency/volume characteristic of micturition in the rat is also gender-specific. Data suggest that the volume voided per micturition depends on the urine production rate. PMID- 11385696 TI - NK2 tachykinin receptors mediate contraction of the pig intravesical ureter: tachykinin-induced enhancement of non-adrenergic non-cholinergic excitatory neurotransmission. AB - The current study was designed to characterize the functionally active tachykinin receptors involved in tachykinin-elicited contractions in the pig intravesical ureter, and to investigate the possible modulation exerted by the natural tachykinins substance P (SP) and neurokinin A (NKA) on the non-adrenergic non cholinergic (NANC) excitatory ureteral neurotransmission. In pig intravesical ureteral strips pretreated with phosphoramidon (10(-5) mol/L) to block the endopeptidase activities, isometric force recordings showed that SP, NKA, and the NK2 receptor selective agonist [beta-Ala(8)]-NKA (4-10), all three induced contractions, with the following potency order: NKA > [beta-Ala(8) ]-NKA (4-10) > SP. [Sar(9), Met(O(2))(11)]-SP and senktide, selective agonists of the NK1 and NK3 receptors, respectively, failed to modify the ureteral tone. Urothelium removal and incubation with tetrodotoxin (10(-6) mol/L), phentolamine (10(-7) mol/L), propranolol (3 x 10(-6) mol/L), atropine (10(-7) mol/L) and indomethacin (3 x 10(-6) mol/L), did not alter the contraction induced by a submaximal (10(-7) mol/L) dose of [beta-Ala(8)]-NKA (4-10). MEN 10,376 (10(-8)-10(-7) mol/L), a NK2 receptor antagonist, reduced the contraction to 3 x 10(-8) mol/L NKA. GR 82334 (10(-6) -10(-5) mol/L) and SR 142801 (10(-8)-10(-7) mol/L), selective antagonists of the NK1 and NK3 receptors, respectively, did not modify that contraction. In pig intravesical ureteral strips in NANC conditions, SP and NKA induced a potentiation of the contractions to electrical field stimulation (EFS) and to exogenous ATP. The results suggest that the tachykinins evoke a direct contraction of pig intravesical ureteral strips through NK2 receptors located in the smooth muscle. SP and NKA exert an enhancement of the NANC excitatory neurotransmission of the pig intravesical ureter. PMID- 11385698 TI - Confidence in the courts: a comparison of users and non-users. AB - Much work has been done on the correlates of confidence in the United States Supreme Court. However, very little research has been undertaken to discern the correlates of confidence in state and local courts. Using survey data from Louisiana, we examine confidence in state and local courts. We focus on the role of experience, arguing that the opportunity for wide participation in these courts makes the confidence calculation different from that of a remote institution like the US Supreme Court. We find that, indeed, experience matters and further, that type of experience matters. Those with more stake in the outcome of the court case and less control over it (e.g., defendants) are least confident in state and local courts, while those with little stake and substantial control (e.g., jurors) are most confident in them. Procedural justice concerns also loom large in the confidence calculation for these lower courts. Timeliness, courtesy, and equal treatment all affect public confidence. PMID- 11385699 TI - Public trust and confidence in legal authorities: What do majority and minority group members want from the law and legal institutions? AB - Discussions of public trust and confidence in the police and the courts often assume that the key to public feelings is the public's evaluation of the outcomes that the public receives from these legal authorities. In the case of the courts, discontent is often assumed to be linked to issues of cost and delay-instrumental concerns about the outcomes delivered to the public by the courts. In the case of the police, the inability to effectively control crime is frequently seen as driving public evaluations. This article presents an alternative procedural justice based model that links public trust and confidence to views about the manner in which legal authorities treat the public. Drawing upon psychological research about public evaluations of institutions and authorities it is argued that the key issue that shapes public views is a process based evaluation of the fairness of the procedures that the police and courts use to exercise their authority. Analyses from several studies exploring the basis of public views support this procedural justice based model of public evaluation. In addition, the results provide suggestions about the elements of procedures that are central to public judgments about their fairness. PMID- 11385700 TI - A satisfied clientele seeking more diverse services: Latinos and the courts. AB - This article assesses Latino views of the court system, both of their beliefs about the courts and their experiences with the courts. Relying primarily on the Latino over-sample of the National Center for State Courts (2000) survey of public attitudes toward the courts, we evaluate three aspects of the Latino judicial relationship. First, we measure whether there are differences between Latino and non-Latino views of the courts. Second, we assess whether there are differences within the Latino community, particularly differences based on nativity, in Latino attitudes toward the courts. Finally, we assess the implications of Latino views of the courts for US society in general. We are particularly concerned about whether having a greater share of Latinos in the population will create new pressures to reform the judiciary. PMID- 11385701 TI - Race, income, and perceptions of the U.S. court system. AB - This article reports on the effect of income within race on African Americans' perception of the courts. Our findings are somewhat consistent with the previous research on black middle-class relative dissatisfaction with various American institutions. That is, unlike whites and Latirios in our study, we find that higher-income African Americans are more skeptical of the notion that blacks receive equal treatment in the courts. This same group also reported less confidence in the court's handling of specific types of cases (e.g., civil, criminal and juvenile delinquency cases.) However, better off blacks were more likely than poor blacks to have confidence in the U.S. Supreme Court and community courts. These findings point a more complex account of African American perceptions of the courts, an account that draws a distinction between diffused and specific support of the courts. PMID- 11385702 TI - Not bound by the law: legal disobedience in Israeli society. AB - The issue of whether there is a 'prima facie obligation to obey the law' has intrigued human society since the days of Socrates. However, most of the writings in this field have dealt with theoretical aspects of the issue, such as the boundaries of legal obedience and frameworks defining the circumstances under which a citizen is not obliged to obey the law. Very few studies have investigated the phenomenon of legal disobedience empirically. The current study is based on a survey of Israeli citizens belonging to three sectors of the population (Jews in the general population, Israeli Arabs, and orthodox Jewish students enrolled in religious yeshiva seminaries). Respondents' attitudes towards the judicial system, the rule of law, and the duty to obey state laws were examined by means of a questionnaire especially designed for the study. The findings point to gaps between the three groups: Compared to the Arab population and the yeshiva students, support for state laws and the rule of law was stronger among Jews in the general population and, conversely, belief in the supremacy of other laws (i.e. religious laws) over state laws and readiness to take the law into one's own hands were stronger among the Arabs and the yeshiva students, compared to the general Jewish population. PMID- 11385703 TI - Generational and age-based differences in attitudes towards jury service. AB - Analysis of surveys of jurors, potential jurors, and the general public show significant differences in attitudes towards jury service by the age of the respondent. This study analyzes the degree to which these differences are the result of generational effects, in which younger citizens are likely to continue in their beliefs about jury service as they age, and the degree to which they are a result of a respondent's life circumstances-income, employment status, or family status-and are thus not likely to be carried with jurors as they age. The article shows that, while there are differences in confidence in the courts by age group, younger jurors are more confident in their own abilities to serve well as jurors but more skeptical of the court as a whole; most differences in attitudes towards jury service are linked to life-cycle phenomena. As such, courts should work to provide assistance to particular age groups within the jury pool, including child care and appropriate compensation, if they are to attract jurors who are representative by age of the general public. PMID- 11385704 TI - Judging jury service: results of the North Carolina administrative office of the courts juror survey. AB - This study examined venirepersons' and jurors' levels of satisfaction with jury service. Surveys were distributed to all persons who reported for state court jury service during two one-week periods in each North Carolina county. Questions concerned satisfaction with various aspects of jury service, the effects of service, hardships experienced, details of and reactions to cases heard, and basic demographic information. Responses were obtained from 82 of the 100 counties and 4,654 venirepersons (of whom 1,478 served as jurors). Consistent with prior research on juror experiences, results generally revealed high levels of satisfaction and positive opinions about various aspects of jury service. Service did not influence opinions about the courts for most respondents, and those whose opinions changed tended to become more positive about jury service. Suggestions for future research emphasize giving higher priority to publishing unpublished research on jury experience, explaining the high satisfaction levels observed in samples of jurors, and examining the impact of trial reform on juror satisfaction levels. PMID- 11385705 TI - Denaturing high-performance liquid chromatography: A review. AB - Denaturing high-performance liquid chromatography (DHPLC) compares two or more chromosomes as a mixture of denatured and reannealed PCR amplicons, revealing the presence of a mutation by the differential retention of homo- and heteroduplex DNA on reversed-phase chromatography supports under partial denaturation. Temperature determines sensitivity, and its optimum can be predicted by computation. Single-nucleotide substitutions, deletions, and insertions have been detected successfully by on-line UV or fluorescence monitoring within 2-3 minutes in unpurified amplicons as large as 1.5 Kb. Sensitivity and specificity of DHPLC consistently exceed 96%. These features and its low cost make DHPLC one of the most powerful tools for the re-sequencing of the human and other genomes. Aside from its application to the mutational analysis of candidate genes, DHPLC has proven instrumental in elucidating human evolution and in the mapping of genes. Employing completely denaturing conditions, the utility of DHPLC has been extended to the genotyping of known polymorphisms by utilizing the ability of poly(styrene-divinylbenzene) to resolve single-stranded DNA molecules of identical size that differ in a single base. Under completely denaturing conditions, it is thus possible to resolve all possible base substitutions with the single exception of C-->G transversions. Improvements in throughput became feasible with the recent introduction of monolithic poly(styrene-divinylbenzene) capillaries that lend themselves to the fabrication of arrays connected to a multi-color laser induced fluorescence scanner or a mass spectrometer. PMID- 11385706 TI - Automation in genotyping of single nucleotide polymorphisms. AB - Automation for genotyping of single nucleotide polymorphisms (SNPs) can be split into the automation of the sample preparation and the automation of the analysis technology. SNP genotyping methods are reviewed and solutions for their automation discussed. A panacea for SNP genotyping does not exist. Different scientific questions require adapted solutions. The choice of a technology for SNP genotyping depends on whether few different SNPs are to be genotyped in many individuals, or many different SNPs are to be genotyped in few individuals. The requirements of throughput and the ease of establishing an SNP genotyping operation are important, as well as the degree of integration. The potential and state-of-the-art of different solutions are outlined. PMID- 11385707 TI - Transthyretin mutations in hyperthyroxinemia and amyloid diseases. AB - Over 80 different disease-causing mutations in transthyretin (TTR) have been reported. The vast majority are inherited in an autosomal dominant manner and are related to amyloid deposition, affecting predominantly peripheral nerve and/or the heart. A small portion of TTR mutations are apparently non-amyloidogenic. Among these are mutations responsible for hyperthyroxinemia, presenting high affinity for thyroxine (a TTR ligand). Compound heterozygotic individuals for TTR mutants have been described; noteworthy is the clinically protective effect exerted by a non-pathogenic over a pathogenic mutation. Current TTR mutations and their significance are briefly reviewed here. PMID- 11385708 TI - Spectrum of ABCA4 (ABCR) gene mutations in Spanish patients with autosomal recessive macular dystrophies. AB - The ABCA4 gene has been involved in several forms of inherited macular dystrophy. In order to further characterize the complex genotype-phenotype relationships involving this gene, we have performed a mutation analysis of ABCA4 in 14 Spanish patients comprising eight STGD (Stargardt), four FFM (fundus flavimaculatus), and two CRD (Cone-rod dystrophy) patients. SSCP (single-strand conformation polymorphism) analysis and DNA sequencing of the coding and 5' upstream regions of this gene allowed the identification of 16 putatively pathogenic alterations, nine of which are novel. Most of these were missense changes, and no patient was found to carry two null alleles. Overall, the new data agree with a working model relating the different pathogenic phenotypes to the severity of the mutations. When considering the information presented here together with that of previous reports, a picture of the geographic distribution of three particular mutations emerges. The R212C change has been found in French, Italian, Dutch, German, and Spanish but not in British patients. In the Spanish collection, R212C was found in a CRD patient, indicating that it may be a rather severe change. In contrast, c.2588G>C, a very common mild allele in the Dutch population, is rarely found in Southern Europe. Interestingly, the c.2588G>C mutation has been found in a double mutant allele together with the missense R1055W. Finally, the newly described L1940P was found in two unrelated Spanish patients, and may be a moderate to severe allele. PMID- 11385709 TI - Frequency of recent retrotransposition events in the human factor IX gene. AB - Two germline retrotransposition mutations of recent origin were observed in 727 independent mutations (0.28%) in the human factor IX gene (F9) of patients with hemophilia B: 1) a 279 bp insertion in exon H originating from an Alu family of short interspersed elements not previously known to be active and, 2) a 463 bp insertion in exon E of a LINE1 element originating in the maternal grandmother. If the rates of recent germline mutation in F9 are typical of the genome, a retrotransposition event is estimated to occur somewhere in the genome of about one in every 17 children born. Analysis of other estimates for retrotransposition frequency and overall mutation rates suggests that the actual rate of retrotransposition is likely to be in the range of one in every 2.4 to 28 live births. PMID- 11385711 TI - The R71G BRCA1 is a founder Spanish mutation and leads to aberrant splicing of the transcript. AB - In a BRCA1 screening in familial breast cancer carried out in different centres in Spain, France, and United Kingdom, a missense mutation 330A>G which results in a Arg to Gly change at codon 71 (R71G) was independently identified in 6 families, all of them with Spanish ancestors. This residue coincides with the -2 position of the exon 5 donor splice site. We further investigated the effect of this base substitution on the splicing of BRCA1 mRNA. The sequence analysis of the cDNA indicated that 22 bp of exon 5 were deleted, creating with the first bases of exon 6 a termination codon at position 64, which results in a truncated protein. The BRCA1 haplotype of the R71G carrier patients and Spanish controls was analysed by use of six microsatellites located within or near BRCA1. Our results are consistent with the possibility that these families shared a common ancestry with BRCA1 R71G being a founder mutation of Spanish origin. PMID- 11385710 TI - Mutations P51U and G122E in retinal transcription factor NRL associated with autosomal dominant and sporadic retinitis pigmentosa. AB - Retinitis pigmentosa (RP) is the most frequent form of inherited retinopathy. RP is genetically heterogeneous with autosomal dominant, autosomal recessive and X linked forms. Autosomal dominant retinitis pigmentosa (adRP) accounts for about 20-25% of all RP cases. At least ten adRP loci have so far been mapped. However, mutations causing adRP have been identified only in four retina-specific genes: RHO (encoding rhodopsin) in approximately 20% of adRP families, peripherin/RDS (3 5% of adRP) and recently RP1 (Pierce et al., 1999, Sulivan et al., 1999) and NRL gene. Only one mutation in the NRL gene causing adRP has so far been reported (Bessant et al., 1999). Here we report a novel mutation Pro51Leu in an adRP Spanish family supporting that mutation in NRL is the cause of adRP. A second missense mutation Gly122Glu has been observed in a simplex RP patient that may represent a sporadic case of retinitis pigmentosa. Hum Mutat 17:520, 2001. PMID- 11385712 TI - Four novel MSH2 and MLH1 frameshift mutations and occurrence of a breast cancer phenocopy in hereditary nonpolyposis colorectal cancer. AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is caused by mutations of genes encoding for proteins of the mismatch repair (MMR) machinery. The majority of mutations occur in the MLH1 and MSH2 genes, and consist of splice-site, frameshift and nonsense changes, leading to loss of protein function. In this study, we screened 7 HNPCC families for MLH1/MSH2 mutations. Sequence changes were identified in 5 families. Four alterations were novel 1- or 2-bp deletions or insertions causing a frameshift and appearance of premature stop codons (MLH1: c.597-598delGA, c.1520-1521insT; MSH2: c.1444delA, c.119delG). The four small insertions/ deletions were located within stretches of simple repeated sequences. By reviewing the HNPCC mutation database, we found that the majority of 1-2 bp frameshift mutations similarly affects simple repetitive stretches, pointing to DNA polymerase slippage during replication as the most likely source of such errors. We also evaluated microsatellite instability (MSI) in a breast carcinoma (BC) from an MLH1 mutation carrier. While a colon cancer from the same individual showed MSI, the BC specimen was MSI-negative, indicating that development of the latter tumor was unrelated to MMR impairment, despite presence of a constitutional MLH1 mutation. Hum Mutat 17:521, 2001. PMID- 11385713 TI - Mutations in the connexin26/GJB2 gene are the most common event in non-syndromic hearing loss among the German population. AB - Congenital sensorineural hearing loss affects approximately 1/1,000 live births. Mutations in the gene encoding connexin26 (GJB2) have been described as a major cause of genetic nonsyndromic hearing impairment. Additionally, another gap junction gene, connexin30 (GJB6), was found to be responsible for hereditary hearing loss. We have studied 134 patients with severe to profound hearing loss or deafness and 13 patients with mild to moderate nonsyndromic sensorineural hearing loss in order to evaluate the prevalence of connexin26 and connexin30 mutations in Germany. Mutations in the connexin26 gene were found in 30 patients (22%) with profound to severe hearing impairment whereas only one novel single nucleotide polymorphism (396G-->A) in the connexin30 gene was detected. Among the 13 patients with mild to moderate hearing loss neither mutations in the connexin26 nor in the connexin30 gene could be detected. These results demonstrate that mutations in the connexin26 gene are also a frequent cause of hereditary non-syndromic hearing loss in Germany. Therefore a screening of mutations in the connexin26 gene should be performed in every case of non syndromic hearing loss of unknown origin. PMID- 11385714 TI - Novel mutations in the emerin gene in Israeli families. AB - Emery-Dreifuss Muscular Dystrophy (EMD or EDMD) is a rare X-linked recessive disorder, characterized by progressive muscle wasting and weakness, contractures, and cardiomyopathy, manifesting as heart block. Mutation analysis at the EMD gene locus was performed in 4 unrelated Israeli families with X-linked EMD and in one sporadic case. In the 4 families 4 different mutations were found, 3 of which were novel. These included two frame shift mutations in exon 2 (333delT and 412insA) and one base pair substitution at the consensus +1 donor splice in intron 5 (1429G-->A). The fourth mutation in exon 6 (1675-1678delTCCG) has been previously described. No mutations were identified in the one sporadic case. Two of the three novel mutations were found in exon 2. A summary of the previously published mutations described in the EMD Mutation Database (http://www.path.cam.ac.uk/emd/) as well as the mutations described in our study suggest that the distribution of mutations in EMD gene is not entirely random and that exon 2 is prone to mutations. Hum Mutat 17:522, 2001. PMID- 11385715 TI - Sequence analysis and transcript identification within 1.5 MB of DNA deleted together with the NDP and MAO genes in atypical Norrie disease patients presenting with a profound phenotype. AB - Mutations at the Norrie disease gene locus, NDP, manifest in a broad range of defects. These range from a relatively mild, late-onset, exudative vitreoretinopathy to congenital blindness and sensorineural deafness combined, in some cases, with mental retardation. In addition, extensive deletions involving the NDP locus, located at Xp11.3, the adjacent monoamine oxidadase genes MAOA and MAOB, and additional material, result in a more severe pattern of symptoms. The phenotypes include all or some of the following; mental retardation, involuntary movements, hypertensive crises and hypogonadism. We extended an existing YAC contig to embrace the boundaries of three of the largest deletions and converted this into four PAC contigs. Computer analysis and experimental data have resulted in the identification of several putative loci, including a phosphatase inhibitor 2-like gene (dJ154.1) and a 250-bp sequence which resembles a homeobox domain (dA113.3), 1.2 Mb and 400 kb respectively from the MAO/NDP cluster. The pattern of expression of dJ154.1 suggests that it may represent an important factor contributing to the complex phenotypes of these deletion patients. Hum Mutat 17:523, 2001. PMID- 11385716 TI - Molecular analysis of phenylketonuria (PKU) in newborns from Texas. AB - This study describes the mutations at the phenylalanine hydroxylase (PAH) locus in patients with the diagnosis of classic PKU (n=18), hyperphenylalaninemia (HPA) variant (n=9) and benign persistent hyperphenylalaninemia (HPA) (n=13) who were identified by the Texas Newborn Screening Program. Blinded studies were done by sequencing of the 13 exons and exon-intron boundaries of the PAH gene in genomic DNA isolated from dry blood spots. Thirty-six different mutations, including 25 missense mutations, six splice mutations, three deletions and two nonsense mutations were detected in 75 of the 80 mutant alleles (94%). The prevalent mutations were R408W (19%), V388M and IVS10nt-11g->a (6% each), Y414C (5%) and H170D, A403V, T380M and IVS7nt1g->a (4% each). Two novel missense mutations were identified in exon 5 (H170D and N167S). There was genotype/phenotype correlation in 33/40 cases (83%). For this population, exons 12, 11, 7, 5 and 8, which carry 78% of the mutations, would have to be screened first. However, the other exons must be studied when either one or no mutations are found in the primary screening. Hum Mutat 17:523, 2001. PMID- 11385717 TI - Identification of an additional allelic variant (XLS) of the human serotonin transporter gene (SLC6A4): -1201Cins66. PMID- 11385718 TI - First mutation in cardiac troponin C, L29Q, in a patient with hypertrophic cardiomyopathy. PMID- 11385719 TI - Sticky tumor cells. PMID- 11385720 TI - Old liver? Grow a new one. PMID- 11385721 TI - Cost analysis and rate setting manual for animal research facilities. PMID- 11385723 TI - Behavioral phenotyping of transgenic and knockout animals: a cautionary tale. AB - Knockout and transgenic mice are extremely useful for behavioral research, especially for linking specific genes with behaviors. The authors present caveats to be aware of when using such mice in research situations. PMID- 11385722 TI - What's your diagnosis? Bilateral teat swelling in a female goat. Pseudopregnancy with hydrometra. PMID- 11385724 TI - Developing a strategy for the rapid identification of genetically altered mice: an olfactory system perspective. AB - The mouse genome contains a wide array of uncharacterized genes that are involved in olfactory system development and cellular function. The author describes how random mutagenesis of these genes combined with rapid screening techniques afford the identification of the particular olfactory-related genes that have been altered. PMID- 11385725 TI - Preliminary results of olfactory testing in rats without deprivation. AB - Although rodents are nocturnal, their behavior is usually tested during the day. The authors present the results of a preliminary study, which suggest that altering the animals' day:night cycle might be the key to eliminating the need for food or water deprivation prior to testing. PMID- 11385726 TI - Comparison of three sedation regimes for spinal anesthesia in sheep. AB - The authors considered three protocols for spinal anesthesia using sheep as a model. An appropriate spinal anesthesia method would obviate the need for general anesthesia in certain surgical approaches. PMID- 11385728 TI - Automated systems for analysis of rodent activity and behavior. PMID- 11385727 TI - A cart cage for transferring macaques, capuchins, and small dogs. AB - A novel mobile monkey transport cart cage allows ease of handling, safety, secure holding, good visual access to the monkeys, room for large macaques, and ease of assembly, all at a modest cost. PMID- 11385729 TI - OLAW clarifies stance on use of alternate members on IACUCs and protocol and program reviews at collaborating institutions. PMID- 11385730 TI - What's your diagnosis? Fever and leukocytosis in a young beagle. Canine juvenile polyarteritis syndrome (beagle pain syndrome). PMID- 11385731 TI - Pathology of mutant animal models 2001. PMID- 11385732 TI - The influence of genetic background on spontaneous and genetically engineered mouse models of complex diseases. AB - In recent years, the use of genetically altered mice as models of complex human disease has revolutionized biomedical research into the genetics of disease pathogenesis and potential therapeutic interventions. Whether a mouse expresses a spontaneous or induced mutation, it is critical to remember that the observed phenotype is not always the direct result of the genetic alteration. The author points out the importance of considering the genetic background of the strain used to create these important models. PMID- 11385733 TI - Commercial cotton nesting material as a predisposing factor for conjunctivitis in athymic nude mice. AB - Environmental enrichment for rodents is beneficial, but compatibility between the enrichment device and the rodent strain must also be considered. The authors present a case in which the use of a specific form of environmental enrichment- cotton bedding material--proved detrimental to the health of athymic nude mice, increasing the likelihood of conjunctivitis. PMID- 11385734 TI - Handling the cotton rat for research. AB - First used as an animal model of poliomyelitis in the late 1940s, the cotton rat is currently used in biomedical research for studies of human respiratory syncytial virus and filariasis. The author provides useful information relevant to the care of these research animals, including husbandry considerations, proper handling, and common laboratory procedures. PMID- 11385735 TI - Background noise. PMID- 11385736 TI - Soy protein alone won't reduce cholesterol--at least not in hamsters. PMID- 11385737 TI - Alzheimer's cure? PMID- 11385738 TI - A shot against cocaine addiction. PMID- 11385739 TI - Public law establishes interagency coordinating committee for the validation of alternative methods. PMID- 11385740 TI - What is your diagnosis? Hair loss in lactating rats. Barbering by damor sucklings. PMID- 11385741 TI - Elements of an occupational health and safety program: deficiencies identified by AAALAC international. AB - An effective occupational health and safety program is critical to ensure personnel safety in working with animals. The authors present data compiled from AAALAC international site visits conducted between 1993 and 1999, which indicate how programs can fall short of current recommendations. PMID- 11385742 TI - Assessment of risks posed by research animals: a working model. AB - In the US, the research animal facility is perhaps the most highly regulated area of interaction between humans and animals. Informing staff members of the risks posed by research animals is one of many expectations that must be met by institutions. The author offers a model for animal risk assessment. PMID- 11385743 TI - Radioactivity and radiation safety: basic concepts for laboratory animal technologists and technicians. AB - Laboratory animals treated with radioactive compounds are a radiation-safety concern. It is important for technologists and technicians who work with these animals to understand ionizing radiation and radiation-safety practices. The author discusses licensing, contamination control, and how to deal with radioactive waste. PMID- 11385744 TI - Unique design for fixed ventilated changing station. AB - The Jackson Laboratory animal colonies present a unique challenge for the design and operation of an animal changing station that maximizes protection of animal health and welfare while also protecting the health and safety of the animal caretaker. The authors describe the modification of a fixed ventilated changing station for improved animal health, reduced ergonomic strain, and decreased allergen exposure. PMID- 11385745 TI - Be a team player. PMID- 11385746 TI - Gaining brain cells. PMID- 11385747 TI - Mixture for memory retention. PMID- 11385748 TI - President Clinton signs the Chimpanzee Health, Improvement, Maintenance, and Protection Act. PMID- 11385749 TI - What's your diagnosis? Pigmented mass in an experimental gerbil. Spontaneous malignant melanoma. PMID- 11385750 TI - An integrated database for managing animal study proposals and animal inventory for the small animal facility. AB - The authors describe how they created a user-friendly, multifunctional database for use by a variety of vivarium staff. PMID- 11385751 TI - An innovative IACUC protocol system. AB - The author describes a web-based system that has decreased paper-work and protocol approval time, while making protocols available for viewing by the research community. PMID- 11385752 TI - A comprehensive, barcoded system for the management of animal information in a research facility. AB - The authors describe an easy-to-use barcode-based animal tracking system that has improved record keeping and data retrieval and proved instrumental in the containment of an outbreak. PMID- 11385753 TI - Software for lab animal facilities. PMID- 11385754 TI - What's your diagnosis? Cardiopulmonary arrest in a Burmese python. Aortic aneurysm. PMID- 11385755 TI - Future directions in small animal imaging. AB - The adaptation and development of imaging technologies for use in small animals has the potential to be a refinement with profound effects on how basic cancer research using animals is conducted. The authors describe how NCI funding is helping to advance research in this area. PMID- 11385756 TI - A review of high-resolution X-ray computed tomography and other imaging modalities for small animal research. AB - Dedicated high-resolution small animal systems have recently emerged as important new tools for laboratory animal research. These imaging systems permit researchers to noninvasively screen animal models for mutations or pathologies and to monitor disease progression and response to therapy. The authors survey various small animal imaging modalities, including MRI, PET, SPECT, and microCT, and discuss several representative microCT mouse imaging studies. PMID- 11385757 TI - Body condition scoring: comparing newly trained scorers and micro-computed tomography imaging. AB - A Body Condition Scoring (BCS) protocol is an easily learned tool that can be used as a means of body condition assessment for random populations of mice. Here, the authors use X-ray computed tomography technology to show that BCS is a quick and effective method for evaluating parameters such as muscle thickness and fat, and that the method is equally as accurate when employed by newly trained or expert scorers. PMID- 11385758 TI - Effects of changes to Data Protection Act. PMID- 11385759 TI - What unit for blood pressure? PMID- 11385761 TI - Pap smear replacement. PMID- 11385760 TI - Fungal treatment approved. PMID- 11385762 TI - Cervical cancer vaccine trials started. PMID- 11385763 TI - Proceedings of the 6th European Symposium on Calcium Binding Proteins in Normal and Transformed Cells. Paris, France, 14-17 June 2000. PMID- 11385764 TI - [Progress of dentistry in the twentieth century 3. Prosthetic dentistry]. PMID- 11385765 TI - [Clinical dentistry--a fine profession is worth a vision]. PMID- 11385766 TI - [A 'difficult' prosthesis patient]. PMID- 11385768 TI - [ARTSENET, website intended not only for physicians]. PMID- 11385767 TI - [Cholera 1]. PMID- 11385769 TI - [Therapeutic strategies for impacted maxillary canines]. AB - A disturbance in the normal eruption pattern of one or both of the maxillary canines is found in 1% to 2% of the teenagers aged ten to thirteen years. Frequently, dentists refer these patients to orthodontists who, in turn, seek the assistance of an oral and maxillofacial surgeon to retrieve the impacted canine, either by exposure alone or by exposure and attachment of a bracket and ligature for orthodontic extrusion. A common request in this procedure is the extraction of the persistent upper deciduous canine and/or, in case of an Angle Class II molar relationship, extraction of the upper first premolar. It has been shown, however, that on average 15% of these exposed or ligated maxillary canines fail to erupt. Therefore, one should be reluctant to extract the deciduous canine or the permanent first premolar before it is clear that the impacted maxillary canine will indeed erupt after surgical exposure and/or placing of a bracket and ligature for orthodontic extrusion. PMID- 11385770 TI - [Overdentures covering natural roots. The use of chlorhexidine gel]. AB - To prevent caries and periodontal disease in overdenture abutment teeth, daily application of chlorhexidine gel is recommended. In order to get insight into the use of the gel, 29 overdenture wearing patients were interviewed. In addition the patients were requested to show the use of the gel. It was concluded that at any time the patients were instructed about the use of the gel. Nevertheless, a majority of the patients did not use the gel daily and in accordance with the instructions given. Application of the gel in the abutment depressions of the overdenture appeared to be a heavy task. PMID- 11385771 TI - [Syndromes 19. Fetal alcohol syndrome]. AB - Foetal alcohol syndrome is a syndrome with a rather high incidence. It is characterized by mental retardation, growth deficiency, a striking facial appearance (a.o. short palpebral fissures, short, upturned nose, hypoplastic philtrum, hypoplastic maxilla). Malocclusion and a disturbed facial growth may occur. Mental retardation can interfere with dental treatment. PMID- 11385772 TI - Carcinoma of the ovary. PMID- 11385774 TI - Carcinoma of the vulva. PMID- 11385773 TI - Carcinoma of the vagina. PMID- 11385775 TI - Gestational trophoblastic diseases. PMID- 11385776 TI - Carcinoma of the corpus uteri. PMID- 11385777 TI - Carcinoma of the cervix uteri. PMID- 11385778 TI - Carcinoma of the Fallopian tube. PMID- 11385779 TI - [Bruxism--from Matteus to Van Dale]. PMID- 11385780 TI - [Bruxism: well known, but difficult to fathom]. AB - Tooth clenching or grinding may cause serious problems for patients and dentists. Symptoms occur at the level of the teeth (wear), the periodontium and the musculoskeletal system. In the past, the definition of bruxism often was vague and controversial, and also a clinical diagnosis appeared to be difficult. More recently research in the sleep laboratory, using a combination of electromyography video- and sound recording, made it possible to distinguish between bruxism and other oral behaviour. The association between pain and dysfunction of the masticatory system with bruxism is still unclear. PMID- 11385781 TI - [Etiology of bruxism: morphological, pathophysiological and psychological factors]. AB - Bruxism is a controversial phenomenon, but there is consensus about the multifactorial nature of the etiology. Besides peripheral (morphological) factors, central (pathophysiological and psychological) factors can be distinguished. In the past, morphological factors, like occlusal discrepancies and the anatomy of the bony structures of the orofacial region, have been considered the main causative factors for bruxism. Nowadays, these factors play only a minor role, if any. Recent focus is more on the pathophysiological factors. For example, bruxism has been suggested to be part of a sleep arousal response. In addition, bruxism appears to be modulated by various neurotransmitters in the central nervous system. More specifically, disturbances in the central dopaminergic system have been linked to bruxism. Further, factors like smoking, alcohol, drugs, diseases, and trauma may be involved in the bruxism etiology. Psychological factors like stress and personality are frequently mentioned in relation to bruxism as well, but research shows controversial results. Taken all evidence together, bruxism appears to be mainly regulated centrally, not peripherally. PMID- 11385782 TI - [Bruxism and overload of periodontium and implants]. AB - Bruxism is responsible for occlusal tooth wear but can not induce nor aggravate gingivitis or periodontitis. Bruxism induces jiggling forces, which cause a clinical tooth hypermobility, radiologically seen as a widened periodontal space. Although there is no direct causal relation between bruxism and implant failure, implant overload may lead to fractures of the components and bone loss. PMID- 11385783 TI - [Bruxism and temporomandibular disorders]. AB - Bruxism is generally regarded as an important predisposing as well as an initiating factor in temporomandibular disorders. Many studies have demonstrated an association between bruxism and symptoms and signs of temporomandibular disorders, in adults as well as in children. However, a causal relationship has not been established. Currently, the most probable hypothesis is that other factors, reducing the adaptive capacity of the temporomandibular joint and associated tissues, have to be present for bruxism to play an initiating role. PMID- 11385784 TI - [Dental and pharmacological treatment options for bruxism]. AB - Bruxism has a multifactorial etiology. The consequences of bruxism, abrasion of teeth and a possible craniomandibular disorder, present themselves periferal. So, treatment can have divers points of action. Counseling seems to be useful in all cases. Treatment with a hard acrylic occlusal appliance (oral splint) is especially useful in those situations where the patient has a lot of abrasion or suffers from a craniomandibular disorder with pain. Pharmacological treatment will have a more central effect. Some drugs are effective on bruxism. However, the real pharmacological mechanisms have not yet been discovered. In the absence of data from double-blind, polysomnographic, controlled studies, pharmacotherapy treatment is only indicated in severe cases on a short-term base. Until new data come available, the dentist has to use conventional therapies only. PMID- 11385785 TI - [Treatment of bruxism: physiotherapeutic approach]. AB - Epidemiological and experimental studies suggest that there is a relationship between bruxism and pain in the orofacial region, and between bruxism and restricted mandibular movements. However, the exact nature of this relationship remains unclear. Therefore, up till now, the following working hypothesis is used: bruxism can lead to (chronic) pain complaints and restricted mandibular movements, when its intensity exceeds the adaptation capacity of the musculoskeletal structures. In that case, the aims of physical therapy treatment are twofold: to decrease symptoms, such as pain ('symptom therapy'), and to teach the patient to recognise and reduce the bruxism ('behavioral therapy'). Techniques used for symptom treatment are massage and stretching exercises. For the behavioral therapy exercises and myofeedback are often used. PMID- 11385786 TI - [Role of the psychologist in the treatment of bruxism]. AB - Although there is a controversy about the importance of psychological factors in the development, enhancement and perpetuation of bruxism and other parafunctional activities, the contribution of a psychologist in the treatment of the individual patient is recommended. This contribution consists of diagnostic examination and treatment, most often a short cognitive behavior modification therapy. Because bruxism is mostly examined and treated in relation to the complaints it might cause, especially CMD-complaints, diagnosis and treatment are mostly directed at both. For the diagnosis a questionnaire is used, including a psychological test, which is followed by one or more interviews. The treatment is directed at learning to recognize bruxism activities, to practice alternative behavior, and to learn to cope better with situations that may lead to an increase of bruxism. PMID- 11385787 TI - [Repair of tooth wear; restorative-prosthetic aspects]. AB - Tooth wear is considered to be the third major threat to dental health in the industrialized parts of the world next to caries and periodontal disease. Bruxism can be a cause for excessive tooth wear. Thanks to the successes in the prevention and early treatment of the infectious dental diseases, dentitions last longer as result of which tooth wear is becoming more and more of a problem in the preservation of a physiologic dentition throughout live. Furthermore there seems to be an increasing tendency for the occurrence of severe tooth wear at very early ages. Early recognition of possible contributing factors for tooth wear by the dental team is of utmost importance to enhance success in treating the problem. Prevention of ongoing damage to the remaining tooth structure must be the starting point of every treatment plan. Repair of tooth wear is in many cases a challenge from a restorative/prosthetic perspective: esthetic and phonetic aspects play an important role along with the restorative and prosthetic choices as well as functional considerations the clinician has to deal with. PMID- 11385788 TI - [Adhesive techniques in the treatment of tooth wear due to bruxism]. AB - Wear due to bruxism is not restricted to those surfaces of the tooth that are loaded by occlusion and articulation. Due to tooth flexure wedge-shaped lesions, so-called abfractions, may develop. In the treatment of wear caused by bruxism, direct composite restorations may play an important role. Composite resin can be used to cover exposed dentin and to restore the incisor and canine guidance. In combination with a nightguard, such a treatment may prevent further wear. PMID- 11385789 TI - [Cervical external root resorption]. AB - Cervical external root resorption is a progressive resorption process occurring at the site of the attached gingiva. It is independent on the condition of the pulp. Prerequisites for its occurrence are a damaged root surface and the presence of vascularized connective tissue. The specific stimulus for the resorption to start is still unknown. In most cases the resorption process is without clinical symptoms; diagnosis is made on radiographs. The prognosis of teeth with cervical external root resorption is low: at the time of screening too much tooth material has often been resorbed to be able to make a restoration. Only limited resorption areas with good surgical accessibility may be successfully treated. Thus, of prime importance is an early screening of the resorption process. Therefore, in interpreting radiographs one should not only look for carious or periodontal decay, but also for areas indicative for cervical external root resorption. Complete radiographs are to be advised in order to screen for possible other resorption sites. PMID- 11385790 TI - [Insurability of periodontal care in 1999]. AB - In this study an overview of dental insurance in the Netherlands for periodontal care is presented. Since the dental insurance reform in 1995, dental care is no longer part of the collective health care system. Patients are therefore obliged to enter the market for private dental insurance. Of the 118 available insurance packages 86 (73%) contain dental care. Sixty eight of these packages (58%) include periodontal care. A minority of the companies (4) include periodontal care in all packages. Fifty two percent of the companies apply additional terms, which reduces the scope of the claims. In this paper it is concluded that a sufficient amount and diversity of insurance packages is present. Since patients may not know about the diversity, insufficient coverage of treatment costs may result. It is concluded that market elements are successful integrated in the field of dental insurance. It has to be shown in the future if this success will lead to an increase of dental and periodontal health of the patients. PMID- 11385792 TI - [Progress of dentistry in the twentieth century 2. General dentistry]. PMID- 11385791 TI - [Syndromes 18. Von Recklinghausen's disease]. AB - Von Recklinghausen's disease (neurofibromatosis 1; NF1) is one of the neurofibromatoses and accounts for about 90% of all cases. Inheritance is autosomal dominant with about 30-50% of cases representing new mutations. Characteristic features for NF1 are six or more cafe-au-lait-spots, neurofibromas, Lisch nodules and axillary freckling. Oral manifestation consists of neurofibromas and intrabony lesions. Due to growth of the oral and facial neurofibromas maldevelopment of the facial skeleton and malocclusion are seen. Surgical correction in young individuals easily leads to recurrence. Contour corrected surgery in grown up individuals is possible. PMID- 11385793 TI - [Dentistry on the threshold of the 21st century]. PMID- 11385794 TI - [Preparation with laser?]. PMID- 11385795 TI - [Kinetic cavity preparation]. PMID- 11385796 TI - [Hygienic measures for preparation and filling]. PMID- 11385797 TI - [Dental schools and continuing dental education: from passive listener to active participant]. PMID- 11385798 TI - [Light on new developments]. PMID- 11385799 TI - [New developments in area of dental adhesives]. PMID- 11385800 TI - [Chronic pain in the infraorbital region; diagnostic use of anesthesia]. PMID- 11385801 TI - [Intestinal infections with Escherichia coli type O157]. PMID- 11385802 TI - [Success of direct pulp capping after caries excavations]. AB - Direct pulp capping is not generally accepted as a routine dental procedure. It is claimed to be an unpredictable procedure with a low success rate. However, some clinical studies do show success of direct pulp cappings. The aim of this study was to make an inventory of the success rate of direct pulp cappings performed by dental students, and of the variables which influenced the outcome. The success rate for the first 18 months after treatment was 73.8%. Only the variable 'type of tooth' showed a statistical significant correlation: front teeth showed a higher percentage of success than premolars and molars. PMID- 11385804 TI - [Dutch Act of Agreement on Medical Treatment. Knowledge among dentists]. AB - After more than 25 years of debate about the legislation of patient rights in the Netherlands, the 'Agreement on Medical Treatment' act was passed by the Dutch government in 1995. This article descibes the amount of knowledge general dental practitioners have on several topics of this law, four years after its introduction. A questionnaire was send to 806 dental practitioners, random drawn from the registers of the Dutch Dental Association. The response-rate was 41.6%. Results show that respondents are well informed about some of the most important topics of this law, such as the requirement to obtain the patient's consent to major dental treatments and the fact that information must be based on the 'patient need standard'. Other rights and duties however, are less known. For example, it is still not well known that for routine treatments the patient's explicit consent is not needed, but can be assumed. Also, knowledge about the minimal period for retaining patient dossiers is scarcely present. Some explanations for this lack of knowledge are discussed, especially the lack of relevance of some topics of this law for the dental practice. PMID- 11385803 TI - [Long-term survival of extensive amalgam restorations]. AB - In cost-effectiveness analysis and contemporary treatment planning strategies, the postponement of placement of cast crowns plays an important role. Extensive Amalgam Restorations (EAR), that involve the rebuilding of cusps and the provision of auxiliary retention, are thought to make this postponement possible. This study reports the long-term survival (100 months) of EAR in a randomized, controlled clinical trial. A total of 300 EAR were placed by 3 operators in molar teeth in which one or more cusps were reconstructed. Five different auxiliary retention methods were used for retention of these restorations. As the purpose of EAR is considered to be twofold (to restore a broken-down molar to acceptably function as an independent restoration and to create a substructure for subsequent crown construction), survival was assessed at different levels. The survival rate of EAR as an independent restoration was 88 +/- 2%. The functional survival rate (as an independent restoration or as a substructure) was 92 +/- 2%. The influence of experimental variables (retention method and operator) and background variables (tooth type, extension of EAR and age of patient) on the survival was analysed using Log Rank and Breslow tests. The analyses revealed that there were no statistically significant influences on the survival rates except for the variable 'age of patient' (p < 0.05). EAR was more prone to failure in the group of older patients than the group of younger ones. PMID- 11385805 TI - [Syndromes 21: Ellis-Van Creveld syndrome]. AB - Patients with the Ellis-Van Creveld syndrome have a short stature. The extremities are often plump and markedly shortened, progressively from the trunk to the fingers and the toes. A bilateral postaxial sixth finger is frequent. The most striking and consistent finding in the mouth is fusion of the middle part of the upper lip to the labial sulcus, resulting in a so-called 'whistling deformity'. Congenitally missing teeth, particularly in the frontal region, are a constant finding too. Teeth are usually small and have conical crowns. Supernummer teeth have also been noted. The oral and maxillofacial surgeon will treat the hypertrophic upper frenulum; the dentist will treat the oligodontia of the frontal region and the conical crowns by means of laminated veneers and etch composite bridgework. PMID- 11385806 TI - [Strange structure on an orthopantomogram]. PMID- 11385807 TI - [Osteoporosis]. PMID- 11385808 TI - [How reliable is medical-dental information on Internet?]. PMID- 11385809 TI - [Question of the month:'Have you had a dental procedure recently?']. PMID- 11385810 TI - Scintigraphic evaluation of lung deposition with a novel inhaler device. AB - A gamma-scintigraphic study was carried out in 13 healthy individuals to compare the lung deposition of budesonide from a novel multidose dry powder inhaler (MDPI; Novolizer) with that from the Pulmicort Turbuhaler, and to assess the degree of flow rate dependence of the Novolizer. Median whole lung depositions for the Novolizer at peak inhaled flow rates of 90, 60 and 45 l/min were 32.0, 25.4 and 19.9% of the metered dose, respectively, compared with 21.4% for the Turbuhaler (peak inhaled flow rate 60 l/min). Patterns of regional lung deposition were similar for all four regimens. These data provided proof of concept in vivo for the novel MDPI device, and demonstrated that it delivers drug to the lungs efficiently. The deposition data enabled drug doses to be used in subsequent clinical trials comparing the Novolizer with the Turbuhaler to be predicted with confidence. PMID- 11385811 TI - Therapeutic equivalence of the Sofotec Novolizer to established standard devices in COPD and asthma. AB - The Novolizer is a breath-actuated multidose dry powder inhaler that was developed as an alternative to the pressurized metered dose inhalers. The efficacy, safety and tolerability of the Novolizer has been compared with those of established inhalational devices for the delivery of salbutamol (Sultanol) in patients with chronic obstructive pulmonary disease and for the administration of budesonide (Pulmicort Turbuhaler) to patients with asthma. In both studies, improvement in forced expiratory volume in 1 s in patients who were randomly assigned to use the Novolizer was comparable with that in patients who used the established devices. Furthermore, no differences in tolerability or safety between the Novolizer and the control devices were observed. Finally, patient acceptance of the Novolizer was high, with the majority of patients who used it stating that they would use the device again. PMID- 11385812 TI - Quality of life and patient satisfaction: two important aspects in asthma therapy. AB - Health-related quality of life and satisfaction with treatment are important and complementary measures of the patient's health care experience. Whereas health related quality of life provides information on the implications and consequences of the disease or its treatment for the patient's functional ability and well being, satisfaction with received medical care has an important influence on compliance with therapy. Studies in patients with asthma have shown that health related quality of life is correlated with symptom control and is a predictor of physician visits. Although few studies of treatment satisfaction using validated questionnaires have been performed, there is evidence that ease of use is an important determinant of patient satisfaction with inhalational devices. Prospective, long-term, naturalistic assessments of patient satisfaction with such devices, performed alongside studies of their efficacy, could help to improve the overall management of patients with asthma. PMID- 11385813 TI - New treatments for asthma: current and future aspects. AB - Inhaled beta 2-agonists continue to be the mainstay of asthma therapy, along with inhaled corticosteroids (ICS). A recent advance has been the recognition that 'add-on' or adjunctive therapies to ICS, such as long-acting beta 2-agonists and leukotriene antagonists, are a superior option to increased doses of ICS. Traditionally, inhaled beta 2-agonists and ICS have been administered via separate devices, but combined administration of a long-acting inhaled beta 2 agonist and an ICS by a single inhaler has recently become available in clinical practice. For the future, biotechnology is providing a number of potential therapies for asthma that are directed against some of the inflammatory mediators, such as immunoglobulin E and interleukin-4 and -5, thought to be involved in the pathophysiology of the disease. These biotechnological therapies may eventually provide the opportunity to tailor treatment to the needs of individual patients, and to manipulate the immune system away from allergic responses. PMID- 11385814 TI - Stability and performance characteristics of a budesonide powder for inhalation with a novel dry powder inhaler device. AB - The design and the functionality of an innovative, multidose dry powder inhaler for the topical administration of drugs to the lung are described. The integrated triple inhalation control system, combined with easy handling, should increase acceptance of the device by patients. In-vitro data on a 200-microgram dose of budesonide powder for inhalation have demonstrated the excellent performance and robustness of this pocket-sized, breath-actuated device. The inhaler was developed for a wide range of drugs, and its patented dispersion system can be used with pelletized pure drugs or powder blends. The multidose dry powder inhaler is already on the market for use with one of the key asthma drugs: budesonide (Budecort 200 Novolizer; Sofotec GmbH & Co. KG, Frankfurt, Germany). PMID- 11385815 TI - [Distraction osteogenesis in maxillofacial surgery]. AB - With distraction osteogenesis (DOG) formation of new bone is initiated by gradual separation of osteotomized bone fragments. Both external and internal distraction devices are available. Since its first application in craniomaxillofacial surgery in the early nineties, developments in distraction osteogenesis have been tremendous. Important advantages of this technique are: lack of a donorsite and its associated morbidity; increase of the volume of the soft tissue envelop; less surgical trauma compared to conventional craniomaxillofacial procedures; and the usability of the technique in growing individuals. Disadvantages of DOG are: the sofar limited experience and follow-up in craniomaxillofacial surgery; and the unknown influences on growth. The technique will gradually find its niche and the general dentist will therefore be increasingly confronted with its specific indications. In this paper the general principles of DOG and a number of indications in craniomaxillofacial surgery are discussed. PMID- 11385816 TI - [Oral lichen planus, amalgam and other restorative materials]. AB - Mucosal lesions in the oral cavity, identical to oral lichen planus (OLP), as a consequence of contact with in particular dental silver amalgam may be due to an allergic reaction to mercury or, possibly, a toxic effect on the oral mucosa. Substitution of amalgam preferably by gold--unless sensitisation to gold exists- often results in remission, which sustains a pathogenic role of amalgam in OLP. Substitution of amalgam should be considered if the mucosal lesions correspond topographically to the restorations and epicutaneous patch testing results in a positive reaction on mercury. PMID- 11385817 TI - [Syndromes 20. Tuberous sclerosis complex]. AB - Tuberous sclerosis complex (TSC) is an autosomal dominantly inherited disease. It is clinically a very variable disorder. Hamartomas can develop in many different organs, including the skin, kidneys, heart and brain. Diagnosis of patients with minimal expression of the disease can be very difficult. The dentist can contribute to the (early) diagnosis of the disease. PMID- 11385818 TI - [Progress in dentistry in the twentieth century 4. Orthodontics]. PMID- 11385819 TI - [Surgically assisted maxillary expansion]. AB - Orthodontic expansion of the maxilla can be facilitated by bone cuts through the zygomatic-alveolar process and in the palate lateral of the nasal septum. Stability of the surgical-orthodontic treatment outcome depends on the equilibrium of forces of the tongue and cheeks (functional envelope). Long term, even lifetime, retention is advised. PMID- 11385820 TI - [Cholera 2]. PMID- 11385821 TI - On the acquisition of temporal conjunctions in Finnish. AB - This study concerns the acquisition of complex sentence structures in Finnish. Specifically, three simultaneous and sequential events were acted out with toys in an elicitation task, and the production of "and," "and then," "when," and "after" were observed. There were 48 children in a cross-sectional design at the age levels 3, 4, 5, and 6 years. Immediately after the complex event was presented, the child was asked the initial request "What happened?" If the child did not produce the whole event spontaneously, she or he was prompted by "What else happened?" Finally, the prompted request "When did X?" was asked (X referring to the second action component of the event). The results showed that prompting better revealed the ability of the children, especially that of the younger ones, to use temporal conjunctions in complex sentences, as well as the delicate interplay of language skills and their flexible use. PMID- 11385822 TI - Segmentation and selection of appropriate Chinese characters in writing place names in Japanese. AB - This paper explores the relation between an unknown place name written in hiragana (a Japanese syllabary) and its corresponding written representation in kanji (Chinese characters). We propose three principles as those operating in the selection of the appropriate Chinese characters in writing unknown place names. The three principles are concerned with the combination of on and kun readings (zyuubako-yomi), the number of segmentations, and the bimoraicity characteristics of kanji chosen. We performed two experiments to test the principles; the results supported our hypotheses. These results have some implications for the structure of the Japanese mental lexicon, for the processing load in the use of Chinese characters, and for Japanese prosody and morphology. PMID- 11385823 TI - Cognitive, linguistic, and social aspects of adults' noun definitions. AB - Low-income, rural adults and middle-income, urban adults provided oral definitions for eight common nouns. Two general issues were addressed: (1) whether the rural adults' definitions would conform to the well-documented Aristotelian form typically found among middle-class, well-educated adults; and (2) whether different definitional types would emerge for two different noun classes, social vs. object nouns. Participants' definitions were examined for conceptual content and linguistic form. Among rural participants, the mean proportion of definitions conforming to the Aristotelian model was .13, contrasted with .69 for the urban participants. Also, rural adults were significantly less likely to cast definitions in the conventional linguistic form than were urban adults. On other measures of definitional skill as well, rural participants demonstrated less mastery. There were no significant differences in definitional form between social and object nouns. Various explanations for the findings are considered. PMID- 11385824 TI - Effects of time pressure on mechanisms of speech production and self-monitoring. AB - The purpose of the present study was to examine effects of time pressure on mechanisms of speech production and self-monitoring. The most widely accepted monitoring theory (Levelt, 1989) suggests that monitoring proceeds through language perception, that is, speech error detection is primarily based on the parsing of one's own inner and overt speech. Twenty-four subjects described visual networks at two different rates (normal and fast). The time pressure manipulation affected a number of temporal characteristics: the error to cutoff and cutoff to repair times were shorter in the fast than in the normal condition. The results indicate that the monitor adjusts its speed of error detection and repair planning to the faster speech output rate. The time pressure manipulation did not affect the accuracy of error detection. The implications for the perception theory of monitoring are discussed. PMID- 11385825 TI - Memory for faces, shoes, and objects by deaf and hearing signers and hearing nonsigners. AB - The visual spatial memory of 15 deaf signers, 15 hearing signers, and 15 hearing nonsigners for shoes, faces, and verbalizable objects was measured using the game Concentration. It was hypothesized that the deaf and hearing signers would require fewer attempts than the hearing nonsigners on the shoes and faces tasks because of their experience of using a visual-spatial language; and, in the case of the Deaf, also possibly, due to a compensatory mechanism. It was also hypothesized that memory for shoes would be more like that for faces than for simple objects. It was also anticipated that there would be no difference between the three group's memories for verbalizable objects. Deaf signers were found to be similar to hearing signers, both of whom were better than hearing nonsigners on the faces and shoes tasks. Generally, performance on the faces and shoes tasks was similar and followed the same pattern for the three groups. The three groups performed at a similar level on the objects task. There were no gender differences. PMID- 11385826 TI - Aspectual constraints in the mental lexicon. AB - This study addresses the question of whether constraints on aspectual semantics play a role in lexical processing. Two universal cognitive constraints are identified: "states cannot be delimited" and "telic predicates cannot be further telicized." The study investigates how these are obeyed in the productive process of perfective preverb and stem combination in Bulgarian. An off-line task ascertains that Bulgarian native speakers have a default semantic interpretation for the preverbs under investigation. A visual lexical decision task shows clear legality effects in nonwords composed of existing preverbs and stems, thereby supporting decompositional approaches. It is argued that, after the process of morpheme search, there must be a process of checking for combinatory felicity of the morphemes activated in the lexical access. PMID- 11385827 TI - Effects of early onset brain injury on the development of cognition and behavior: introduction to the special issue. AB - The effects of brain injury acquired early in life on the development of cognition and behavior are not well understood. Deciphering these effects and modeling their neurodevelopmental trajectories are major concerns for clinicians and scientists. Historically, a prevailing notion has been that early-onset brain damage has a more favorable prognosis than does brain damage acquired in adulthood. However, there is growing evidence suggesting that early-onset damage to prefrontal brain structures may have devastating consequences on the emergence of adaptive behavior throughout development. Particularly prominent are disorders of personality, social behavior, and executive functions such as planning and decision making. This special issue presents a series of new empirical studies that address these issues in depth, from several different perspectives, and in both human and animal participants. The findings promise to shed further light on both the neurobiology of development, and diverse neurodevelopmental disorders. Such advances may also enhance clinical diagnosis and facilitate the design of more effective interventions to help reduce the tremendous burden that neurodevelopmental disorders place on personal well-being, family structure, educational systems, and social resources. PMID- 11385828 TI - Long-term sequelae of prefrontal cortex damage acquired in early childhood. AB - Frontal lobe dysfunction is often invoked as a contributing factor in developmental disorders characterized by chronic maladaptive behavior, but interpretation of relevant neuropsychological findings has been hampered by the limited information available regarding the consequences of focal prefrontal damage early in life. We describe here the long-term behavioral and cognitive sequelae of damage to prefrontal cortex in two young adult patients who had sustained their brain damage prior to 16 months of age. In the context of normal neurological examinations, both cases had remarkable histories of impaired decision making, behavioral dyscontrol, social defects, and abnormal emotion. Performances were primarily normal on a broad range of neuropsychological measures (intellect, memory, language, academic achievement, visual perception, and visuoconstruction), but selective impairments of executive function were evident. Early dysfunction in the prefrontal region may result in severe and chronic social maladjustment despite largely normal cognitive abilities. These findings can help inform neuropsychological evaluation of patients with possible prefrontal dysfunction in the setting of developmental disabilities or early brain trauma. PMID- 11385829 TI - Direct and indirect effects of prenatal alcohol damage on executive function. AB - Patients with Fetal Alcohol Syndrome (FAS) and Fetal Alcohol Effects (FAE) often have difficulty functioning appropriately in everyday life and seem to employ poor problem-solving strategies. Tests of executive function are relevant for quantifying the functional deficits and underlying real-life problems associated with prenatal alcohol exposure. This study considers two pathways for the effects of prenatal alcohol on executive function: a direct effect and an indirect effect through prenatal alcohol's effect on IQ. We compared 30 men who had been diagnosed with FAS or FAE with young adults participating in a longitudinal prospective study (n = 419) and 15 control participants that comprised a comparison group. This study is unique in its analysis of the same battery of assessments of executive function in both a large low dose longitudinal study sample and a clinically diagnosed group. Participants were evaluated on 9 tests (including 58 scores) of executive function. For some but not all of the tests in this executive function battery, the decrement in the alcohol exposure group is greater than would be predicted from their IQ scores. We found that 3 of 6 Stroop scores, 2 of 4 Trails scores, 9 of 16 Wisconsin Card Sorting scores, 1 of 2 Ruff's Figural Fluency scores, and 2 of 4 Consonant Trigrams scores appear to be particularly sensitive to the direct effects of prenatal alcohol damage for patients with FAS and FAE. The findings suggest that these executive function tests would be particularly useful in clinical evaluations of persons suspected of fetal alcohol damage because they would not simply reflect deficits in IQ or facial stigmata. PMID- 11385830 TI - Acquired personality disturbances associated with bilateral damage to the ventromedial prefrontal region. AB - Personality changes in 7 participants with bilateral ventromedial prefrontal lesions (PF-BVM), 14 participants with prefrontal lesions but not bilateral ventromedial involvement (PF-NBVM), and 36 with nonprefrontal lesions (NPF) were investigated with the Iowa Rating Scales of Personality Change. Informants rated 30 specific characteristics for degree of disturbance and change from premorbid personality. PF-BVM participants showed a higher rate of acquired disturbances than NPF participants in blunted emotional experience, apathy, low emotional expressiveness, inappropriate affect, poor frustration tolerance, irritability, lability, indecisiveness, poor judgment, social inappropriateness, lack of planning, lack of initiation and persistence, and lack of insight. Differences between the PF-BVM and PF-NBVM groups were significant for several of these characteristics. All 7 PF-BVM participants developed a syndrome, including general dampening of emotional experience; poorly modulated emotional reactions; defective decision making, especially in the social realm; impaired goal-directed behavior; and striking lack of insight. Similarities between this syndrome of "acquired sociopathy" and developmental psychopathy in characteristic personality disturbances and psychophysiological abnormalities suggest that diminished emotionality, impaired decision making, and psychosocial dysfunction may be related to ventromedial prefrontal dysfunction in both groups. PMID- 11385831 TI - Metacognition following pediatric traumatic brain injury: a preliminary study. AB - Metacognition is one of the cognitive processes included under the general term executive functions. The executive functions are widely held to be under the control of the prefrontal cortex, an area often damaged after severe traumatic brain injury (TBI). We examined the metacognitive processing of a group of 9 children with TBI, and a group of 9 healthy, age-matched control children. Children with TBI showed significant impairments in their accuracy of prediction of the ease with which an item would be learned and their ability to predict recall of an item after a 2-hr delay. No significant differences in recall performance between the TBI and control groups were exhibited. The results are interpreted as suggesting an impairment in metacognitive processing resulting from frontal lobe damage after TBI in children. Additional research is necessary to confirm the relation of frontal lobe pathology and severity of injury to metacognitive impairments. PMID- 11385832 TI - Effects of neonatal inferior prefrontal and medial temporal lesions on learning the rule for delayed nonmatching-to-sample. AB - The ability of rhesus monkeys to master the rule for delayed nonmatching-to sample (DNMS) has a protracted ontogenetic development, reaching adult levels of proficiency around 4 to 5 years of age (Bachevalier, 1990). To test the possibility that this slow development could be due, at least in part, to immaturity of the prefrontal component of a temporo-prefrontal circuit important for DNMS rule learning (Kowalska, Bachevalier, & Mishkin, 1991; Weinstein, Saunders, & Mishkin, 1988), monkeys with neonatal lesions of the inferior prefrontal convexity were compared on DNMS with both normal controls and animals given neonatal lesions of the medial temporal lobe. Consistent with our previous results (Bachevalier & Mishkin, 1994; Malkova, Mishkin, & Bachevalier, 1995), the neonatal medial temporal lesions led to marked impairment in rule learning (as well as in recognition memory with long delays and list lengths) at both 3 months and 2 years of age. By contrast, the neonatal inferior convexity lesions yielded no impairment in rule-learning at 3 months and only a mild impairment at 2 years, a finding that also contrasts sharply with the marked effects of the same lesion made in adulthood. This pattern of sparing closely resembles the one found earlier after neonatal lesions to the cortical visual area TE (Bachevalier & Mishkin, 1994; Malkova et al., 1995). The functional sparing at 3 months probably reflects the fact that the temporo-prefrontal circuit is nonfunctional at this early age, resulting in a total dependency on medial temporal contributions to rule learning. With further development, however, this circuit begins to provide a supplementary route for learning. PMID- 11385833 TI - Cortical plasticity and the development of behavior after early frontal cortical injury. AB - It has been known for over 100 years that frontal lobe injury in children is often associated with considerably more functional recovery than after similar injury in adulthood. Systematic study of frontal cortical injury in laboratory animals has shown that this recovery is tightly tied to developmental age: There is a brief window of time during cortical development during which the brain is able to compensate. Simply being young is not sufficient because injury prior to this critical period leads to miserable behavioral outcomes. For humans, the least favorable time for cortical injury is likely at the end of the gestational period, perhaps including the 1st month or so of life whereas the most favorable time is around 1 to 2 years of age. In addition to age, the extent of behavioral recovery is influenced by age at assessment, the nature of the behavioral assessment, sex, and lesion size. Anatomical studies have shown that functional recovery following early cortical injury is correlated with a reorganization of remaining cortical circuitry, including increased dendritic arborization and increased spine density. Recovery, and the compensatory anatomical changes, can also be potentiated by application of different treatments including behavioral therapy, trophic factors, and neuromodulators. Finally, there is preliminary evidence in laboratory animals to suggest that it may be possible to induce neural regeneration in the injured brain and that the regenerated brain functions to support functional recovery. PMID- 11385834 TI - An intervention for children with disorders of executive functions. AB - This article describes a cognitive-behavioral approach to teaching metacognitive executive thinking strategies to children with disorders of executive function. The intervention is based on the notion that some children with disorders of executive function have disorders of higher level language, which predispose them to the executive impairments. It is proposed that teaching and reinforcing metacognitive thinking strategies may help advance verbal mediation of complex tasks and self-regulation of behavior. Despite the growing literature on developmental executive disorders, little has been written about interventions that may enable the children to acquire some of the requisite adaptive skills. The ideas expressed herein should be considered an invitation for the initiation of empirical studies of intervention and outcome effects. PMID- 11385835 TI - Hybrid neural modelling of an anaerobic digester with respect to biological constraints. AB - A hybrid model for an anaerobic digestion process is proposed. The fermentation is assumed to be performed in two steps, acidogenesis and methanogenesis, by two bacterial populations. The model is based on mass balance equations, and the bacterial growth rates are represented by neural networks. In order to guarantee the biological meaning of the hybrid model (positivity of the concentrations, boundedness, saturation or inhibition of the growth rates) outside the training data set, a method that imposes constraints in the neural network is proposed. The method is applied to experimental data from a fixed bed reactor. PMID- 11385836 TI - Automatic buffer capacity model building for the purpose of water quality monitoring. AB - In this paper, buffer capacity profiles are used in the framework of automatic monitoring of water quality. The aim of the proposed methodology is to automatically and stepwise build buffer capacity models for each particular titrated sample, and to quantify the individual buffer systems that constitute the total buffer capacity. An automatic and robust model building algorithm has been developed and applied to many titration curves of effluent and river water samples. It is illustrated that the application of automatically built buffer capacity models mostly results in similar or better estimations of ammonium and ortho-phosphate in the samples compared to a priori fixed buffer capacity models. The automatic modelling approach is also advantageous for alarm generating purposes on e.g. river waters, because unexpected buffers are easily detected. PMID- 11385837 TI - Software sensor design for COD estimation in an anaerobic fluidized bed reactor. AB - In this paper, a method for unknown input estimation in stochastic system is presented. A key problem in bioprocess systems is the absence, in some cases, of reliable on-line measurements for real time monitoring applications. In this paper, a software sensor for an anaerobic digester is presented. Unmeasured components of the influent are estimated from available on-line measurements. Unknown input Kalman filter is discussed to estimate the state and unknown input of the process. First, the theory of unknown inputs optimal filtering in the stochastic case is exposed and a design procedure is proposed. The observer is applied to an anaerobic fluidized bed reactor to estimate the variations in Chemical Oxygen Demand (COD) concentration and experimental results are presented. PMID- 11385838 TI - Global predictive real-time control of Quebec Urban Community's westerly sewer network. AB - Quebec Urban Community (QUC) has selected Global Predictive Real-Time Control (GP RTC) as the most efficient approach to achieve environmental objectives defined by the Ministry of Environment. QUC wants to reduce combined sewer overflows (CSOs) frequency to the St Lawrence river to two events per summer period in order to reclaim the use of Jacques-Cartier Beach for recreational activities and sports of primary contact. QUC's control scheme is based on the Certainty Equivalent Control Open Loop Feedback (CEOLF) strategy which permits one to introduce, at each control period, updated measurements and meteorological predictions. A non-linear programming package is used to find the flow set points that minimise a multi-objective (cost) function, subjected to linear equality and inequality constraints representing the physical and operational constraints on the sewer network. Implementation of GP-RTC on QUC's westerly network was performed in the summer of 1999 and was operational by mid-August. Reductions in overflow volumes with GP-RTC compared to static control are attributed to the optimal use of two existing tunnels as retention facilities as well as the maximal use of the wastewater treatment plant (WWTP) capacity. PMID- 11385839 TI - Operational control of storm sewage at an activated sludge process. AB - Operational control of storm sewage at a wastewater treatment plant has attracted intensive concern over the last decade in the context of river basin management. The focus is on the exploitation of the full capacity of the wastewater treatment plant in attenuating storm sewage, and minimizing a direct storm sewage bypass to the river. Attention is particularly paid to the surge of storm water on the activated sludge process. Based on two typical rain events, this paper discusses the performance of several practical controllers in achieving an optimal effluent performance under storm loadings, without risking internal biomass stability and sludge overflow. The control algorithms tested include various controls of recycle rate, step-feed and step-sludge. Prediction errors of influent characteristics and process responses are also under consideration in the assessment. The results illustrate well the desirability, effectiveness and robustness of the tested controllers. PMID- 11385840 TI - Parameter optimisation of real-time control strategies for urban wastewater systems. AB - Real-time control (RTC) of wastewater systems has been a topic of research and application for over two decades. Attempts so far have mainly focused on one of the parts of the urban wastewater system: either the sewer system, or the treatment plant or the river. Approaches to integrate these subsystems and considering them jointly for control purposes have been pursued only recently. Control of the systems aims at pursuing one (or several concomitant) objectives, which are expressed, for example, in terms of overflow volumes, loads, effluent concentrations, receiving water quality or monetary costs, to name just a few. This paper provides a general and formal definition of the problem to define a real time control algorithm for a given urban wastewater system. A general mathematical optimization problem is formulated, which describes the task of finding an (in some sense) optimum control algorithm. Since this optimization problem is, in the general case, highly non-linear with only limited information available about the objective function itself, optimization methods appropriate for this type of problem are identified. Here, the similarity of the problem to find a control algorithm and of the parameter estimation problem common in mathematical modelling becomes apparent. Hence, methods (and problems encountered) in parameter estimation can be transferred to the problem of determining optimum RTC algorithms. This parallelism is outlined in the paper. As an application of the parameterisation and optimization of control strategies, integrated control of an urban wastewater system is discussed. Since the analysis of integrated control as just described poses certain requirements on a simulation engine, a novel modelling tool, called SYNOPSIS, is utilized here. This simulation tool, comprising of modules simulating water quantity and quality processes in all parts of the urban wastewater system, is embedded into a suite of optimization procedures. An integrated RTC algorithm for the urban wastewater system is formulated, the parameters of which are optimized using various global optimization routines. Comparison of their efficiency indicates good performance for the Controlled Random Search and for the genetic algorithms. The findings suggest that integrated control can indeed lead to an increase in performance of the urban wastewater system. These results appear to be encouraging and justify further work. Areas for further development are identified in the final section of the paper. PMID- 11385841 TI - Supervisory control of wastewater treatment plants by combining principal component analysis and fuzzy c-means clustering. AB - In this paper a methodology for integrated multivariate monitoring and control of biological wastewater treatment plants during extreme events is presented. To monitor the process, on-line dynamic principal component analysis (PCA) is performed on the process data to extract the principal components that represent the underlying mechanisms of the process. Fuzzy o-means (FCM) clustering is used to classify the operational state. Performing clustering on scores from PCA solves computational problems as well as increases robustness due to noise attenuation. The class-membership information from FCM is used to derive adequate control set points for the local control loops. The methodology is illustrated by a simulation study of a biological wastewater treatment plant, on which disturbances of various types are imposed. The results show that the methodology can be used to determine and co-ordinate control actions in order to shift the control objective and improve the effluent quality. PMID- 11385842 TI - Application of mathematical tools to improve the design and operation of activated sludge plants. Case study: the new WWTP of Galindo-Bilbao. Part I: Optimum design. AB - This paper presents a mathematical formulation for the optimum design of a new activated sludge WWTP. The WWTP optimum design problem has been formulated as a Mathematical Programming problem, which is solved through a nonlinear optimisation method. The plant model has been based on the ASM1. The minimum volume of the biological reactors and the minimum total cost (including construction and exploitation costs) have been considered as optimisation criteria. Some practical results are also included, using as a case study the design of the second stage of the Galindo-Bilbao WWTP. PMID- 11385843 TI - Application of mathematical tools to improve the design and operation of activated sludge plants. Case study: the new WWTP of Galindo-Bilbao. Part II: Operational strategies and automatic controllers. AB - This paper presents a new sensitivity analysis methodology for Activated Sludge WWTPs. It is based on both (a) the calculation of the range of "manipulated input variables" that satisfy the restrictions imposed on the "output variables" and (b) on the computation of isolines of the output variables inside the feasible operating space. This analysis allows a more precise description of the operating constraints, facilitates the understanding of the steady-state behaviour of the process and detects possible areas where the process is very sensitive to small disturbances. The feasible operating space for two Activated Sludge WWTP processes for CN removal (RDN, DRDN), using SRT and DO level as "input variables" as well as effluent quality and exploitation costs as the main "output variables" is studied. The proposed methodology facilitates the selection of the appropriate operational strategy and the design of automatic controllers. Some examples of the application of this methodology for the design of automatic controllers in a real WWTP are briefly presented. PMID- 11385844 TI - Advanced monitoring and control of anaerobic wastewater treatment plants: software sensors and controllers for an anaerobic digester. AB - A mass balanced based model representing the dynamical behaviour of anaerobic digester has served as a basis for the design of software sensors for the concentration of inorganic carbon, alkalinity and volatile fatty acids. The predictions of the sensors are close to the actual off-line measurements. The model has also been used to design a model-based adaptive linearizing controller and a fuzzy controller whose objective is to regulate the ratio of the intermediate alkalinity over the total alkalinity below some desired value (0.3) under which the process is assumed to remain in stable conditions and avoid VFA accumulation. Both controllers were calibrated via extensive numerical simulations and implemented. The controllers proved successful in maintaining the ratio of TA over PA below 0.3, even in presence of large variations of the organic load. PMID- 11385845 TI - Advanced monitoring and control of anaerobic wastewater treatment plants: fault detection and isolation. AB - In this paper, a fault detection and isolation approach using fuzzy logic is described for on-line analysis of problems occurring in anaerobic digestion processes. The measurements available on the process are preprocessed to build a vector of fault residuals indicating the magnitude of the problems. This vector is classified into a prespecified category (i.e., a class) which is a state of the system, according to discrimination fuzzy rules. Three different types of classes were defined in a hierarchical structure: sensors faults, sub-process faults and process faults. This approach was developed to handle in real time both technical and biological problems. Demonstration of the practical interest of this study was made using real life experiments and large improvement of the reliability and safety of the process was obtained, thus optimizing the overall wastewater treatment. PMID- 11385846 TI - A new approach towards modelling of the carbon degradation cycle at two-stage activated sludge plants. AB - A pilot plant has been operated in order to investigate the performance and operating characteristics of the plant concept developed for the extension of the main Vienna STP. Due to the different operational modes included in the plant concept, modelling of the carbon degradation becomes of crucial importance. A new activated sludge model is introduced which combines parts of the carbon degradation model concepts as they have been released in the ASM1-model and the ASM3-model, respectively. A method is presented which utilises results from mass balance calculations and sludge stabilisation experiments to reduce the uncertainty in the determination of the values of the simulation model parameters. PMID- 11385847 TI - Advanced monitoring and control of anaerobic wastewater treatment plants: diagnosis and supervision by a fuzzy-based expert system. AB - A fuzzy-based expert system (ES) for the diagnosis and supervision for anaerobic digesters is presented. The system was developed in a Microsoft Windows support using fuzzy logic inference together with a rule base for the implementation of expert knowledge. The ES runs on-line through three main modules, which determine the state and trend of the process, and the best set points for the actuation on the final control elements of the plant. Two further modules run in parallel, when they are required by the operator, using off-line and on-line information for the detection of inhibition due to toxic compounds in the process and for the validation of the on-line diagnosis. The diagnosis and supervision ES was tuned up in order to adjust the membership functions describing the process, and lately tested, running on-line, to study the response of the rule base. PMID- 11385848 TI - System analysis for optimal control of a wastewater treatment benchmark. AB - The paper presents an analysis and optimisation of a wastewater treatment benchmark. The benchmark is a simulation environment defining a plant layout, simulation model, influent data, test procedures and evaluating criteria that should be used for comparing different control strategies. In this paper an analysis of the benchmark which addresses the influences of potential manipulated variables on control performance under different operating conditions is presented. In the study optimisation is used to define the optimal values of the manipulated variables under constant as well as dynamic influent conditions. The results indicate that such an analysis and optimisation give important information about the manipulated variables under varying influent conditions and consequently about possible control strategies. PMID- 11385849 TI - Closed-loop identification and control application for dissolved oxygen concentration in a full-scale coke wastewater treatment plant. AB - The objective of this paper is to apply a closed-loop identification to actual dissolved oxygen control system in the coke wastewater treatment plant. It approximates the dissolved oxygen dynamics to a high order model using the integral transform method and reduces it to the first-order plus time delay (FOPTD) or second-order plus time delay (SOPTD) for the PID controller tuning. To experiment the process identification on the real plant, a simple set-point change of the speed of surface aerator under the closed-loop control without any mode change was used as an activation signal of the identification. The full scale experimental results show a good identification performance and a good tracking ability for set-point change. As a result of improved control performance, the fluctuation of dissolved oxygen concentration variation has been decreased and the electric power saving has been accomplished. PMID- 11385850 TI - Model system for the management of nitrogen leaching at the scale of river basins and regions. AB - In the framework of the EU-project RANR (Regional analysis of subsurface nitrogen retention and its impact on the nitrogen export from land to sea) a model system was developed to estimate groundwaterborne nitrogen inputs into river systems. The core of this model system is composed of a soil nitrogen leaching model (SOIL N) and a groundwater residence time/denitrification model (WEKU). The application of the model system was carried out for the study catchment areas of the Uecker basin (ca. 2400 km2, Germany) and the Gjern basin (ca. 200 km2, Denmark). For both catchment areas, the modelled average nitrogen loads leached into the groundwater were about 40 kg N/ha a, while the remaining groundwaterborne nitrogen intake to rivers was quantified to an average of about 2 kg/ha a. The comparision with observed groundwaterborne riverine nitrogen loads showed a very good agreement, proofing the key role nitrogen retention in groundwater plays in the two catchment areas. With regard to the generalisation and transfer of the SOIL-N/WEKU model concept we assume that the model can be applicated in catchment areas in the European Pleistocene Lowland, which ranges from the Netherlands in the west to the Baltic States and the Ukraine in the east. PMID- 11385851 TI - A georeferenced river quality model. AB - Water quality models have reached a high degree of sophistication, but their weak side remains user interface and output georeferencing. The aim of this paper is to propose an interfacing procedure between two widespread but specialised programming environments: ArcVIEW as a Geographical Information System (GIS) and Matlab as a scientific programming tool for numerical analysis. The proposed solution is based on a Dynamic Data Exchange (DDE) between the two programs in order to operate a Matlab-based water quality model from within the GIS environment. It is shown how special GIS objects must be created and how they operate to achieve the goal of having quality data created by the model placed on a geographical map, together with other site features. PMID- 11385852 TI - Georeferenced regional simulation and aquatic exposure assessment. AB - A simulation model system of the waste water pathways and the aquatic fate assessment is coupled to a discrete digitized river network. Temporal concentration distributions of down-the-drain chemicals are calculated from variable and uncertain input data and compared to monitoring data. The accuracy of the predictions are within a factor of three for the studied pilot catchments. Spatial distributions of the whole catchment give stochastic information on a regional basis for probabilistic risk assessments. PMID- 11385853 TI - Ecological concern as a factor in the optimal attenuation of diverse stressor sources in a stream. AB - The use of an objective function based on fuzzy ecological effect expectation in a genetic optimisation algorithm to obtain site or situation specific stressor attenuation values for the management of diverse stressors emanating from several sources is investigated. The approach is based on the premise that both regulator and regulatee are able to formulate their goals in fuzzy terms. In the case of the regulator the goals will be formulated in terms of acceptability of levels of ecological concern (a fuzzy analogy to ecological risk). In the case of the regulatee it will be formulated in terms of acceptability of the level of attenuation, which is also the control variable. A hypothetical catchment is used to illustrate the principle. PMID- 11385854 TI - Development of an effluent discharge policy for the Tay Estuary based on a finite element model. AB - The tidal hydrodynamics and effluent distribution in estuaries involve a complicated range of solute transport phenomena modelled by partial differential equations. Therefore, the quantitative estimation of the risks of water and soil contamination of coastal areas as a result of polluted estuary flows, or effects of the effluent input on the chemical loads, involves the solution of these equations. Generally, the pollutants load in an estuary is determined by the nature of land use which by altering the watershed hydrology or chemical detention/release in the river banks affect the water quality of the estuaries. The present modelling work aims to investigate the solute transport behaviour in the Tay Estuary in Scotland. Based on this study, an attempt to devise an estuary specific discharge strategy for the Tay has been made. The numerical calculations are based on using 2D Galerkin finite element discretisation of the governing equations in an Eulerian co-ordinate system. The flexibility of the formulation allows it to be extended to moving boundary situations encountered in most tidal water systems. PMID- 11385855 TI - Use of fuzzy method to estimate river nutrient loads from scarce observation. AB - Evaluation of data time series in order to get information about water systems is one of the routinely needed tasks. The results are always associated with uncertainties, of which one arises from data scarcity. Traditional methods, such as regression analyses etc. become rapidly useless with decreasing number of data available. A method based on fuzzy set theory was applied to get more reliable information about the system from scarce databases. Monitored daily flow and water quality data of the medium size Zala River in Hungary were considered as elements of fuzzy sets. Fuzzy rules were generated from data pairs (flow, suspended solids concentration, water temperature and phosphorus load as inputs and output, respectively) from which combined rule bases were set up. These rule bases can be considered as a tool of mapping from the input space to the output space using defuzzification procedure. The method is trainable: it can learn from observations. It is demonstrated that the method is capable to generate daily phosphorus loads and annual balance with acceptable accuracy when it is trained only by weekly, biweekly or monthly data pairs. In comparison to other approaches the tool is well suited to utilize better the information content of scarce observations. Furthermore, monitoring costs can be considerably decreased without substantial information loss since sampling of expensive and labour intensive parameters can be reduced. PMID- 11385856 TI - Developing a model based decision support tool for the identification of sustainable treatment options for domestic wastewater. AB - To enable decision makers to select sustainable wastewater treatment systems, insight into the sustainability of a wide variety of systems should be provided in a transparent way leaving room for adaptation and interpretation according to the local situation. To provide this insight a structured methodology comparing wastewater treatment systems with respect to sustainability is defined. Similar to life cycle assessment (LCA) three phases can be distinguished: (1) goal and scope definition, (2) inventory analysis, and (3) optimisation and results. In the goal and scope definition we set the system boundaries to include most of the water cycle and part of the food cycle. Furthermore, we defined a multi disciplinary set of sustainability indicators including technical, economic, environmental, and socio-cultural aspects. In the inventory analysis these sustainability indicators are quantified using simple static models of wastewater unit operations. Selection of unit operations results in a model of a complete wastewater treatment system. In the optimisation phase the decision maker can weigh the different sustainability indicators and select sustainable options through integer programming. PMID- 11385857 TI - Identification of model structure for aquatic ecosystems using regionalized sensitivity analysis. AB - The Regionalized Sensitivity Analysis (RSA) was developed in 1978, for identifying critical unknown processes in poorly defined systems, thus directing the focus of further scientific investigations. Here, we demonstrate its application to model structure identification, by ranking the constituent hypotheses and identifying the critical elements for progressive revision of the model. Our case study is Lake Oglethorpe--a small monomictic impoundment in South eastern Georgia, USA. Recent studies indicate that the warm temperate regional climate affords an extended growing season--typically from March to October- which promotes bacterial productivity in the lake. The result is a summer food web dominated by microbial processes, in contrast to the conventional phytoplankton-dominated food chains typically observed in the cold temperate lakes of Europe and North America. Starting with a simple phytoplankton-based food web model and a qualitative definition of system behaviour, we use the RSA procedure to establish the critical role of bacteria-mediated decomposition in Lake Oglethorpe, thus justifying the inclusion of microbial processes. Further analysis reveals the importance of size-dependent selective consumption of phytoplankton and bacteria. Finally, we discuss important practical implications of this novel application of the RSA regarding sampling efficiency and statistical robustness. PMID- 11385858 TI - Improving uncertain nutrient load estimates for Lake Balaton. AB - Annual nutrient loads have been estimated for Lake Balaton over three decades. Tributaries may transport about half of the loads into the lake. The contribution of diffuse sources may reach two thirds of the total load. Biweekly/monthly water quality monitoring on small inflows (0.01 m3/s-0.3 m3/s range) results in a high uncertainty of load estimates. This paper evaluates the degree of uncertainties by using analytical expressions of sampling theory. Load-flow relationships were derived for five streams and annual total phosphorus load was predicted by four load estimation methods. A seasonal regression model, based upon the evaluation of historical set of observed phosphorus loads, appeared best to refine load estimates on small inflows. Correction frequently led to load estimates that exceeded uncorrected loads by a factor of two to three. Since the dynamics of the watercourses determined the errors of load estimates, stratified sampling is needed to decrease the uncertainties. PMID- 11385859 TI - Development of a risk assessment based technique for design/retrofitting of WWTPs. AB - Up to now, within the design/retrofit of wastewater treatment plants (WWTPs), deterministic models were used to evaluate different scenarios on their merits in terms of effluent compliance. This paper describes an approach in which a Monte Carlo engine is coupled to a deterministic treatment plant model, followed by risk interpretation in the form of concentration-duration-frequency (cdf) curves of norm exceedance. The combination of probabilistic modelling techniques with the currently available deterministic models allows to determine the probability of exceeding the effluent limits of a WWTP. This percentage of exceedance is accompanied by confidence intervals resulting from the inherent uncertainty of influent characteristics and model parameters. The approach is illustrated for a hypothetical case study, consisting of a denitrifying plant model inspired by the benchmark model described by Spanjers et al. PMID- 11385860 TI - Adequate model complexity for scenario analysis of VOC stripping in a trickling filter. AB - Two models describing the stripping of volatile organic contaminants (VOCs) in an industrial trickling filter system are developed. The aim of the models is to investigate the effect of different operating conditions (VOC loads and air flow rates) on the efficiency of VOC stripping and the resulting concentrations in the gas and liquid phases. The first model uses the same principles as the steady state non-equilibrium activated sludge model Simple Treat, in combination with an existing biofilm model. The second model is a simple mass balance based model only incorporating air and liquid and thus neglecting biofilm effects. In a first approach, the first model was incorporated in a five-layer hydrodynamic model of the trickling filter, using the carrier material design specifications for porosity, water hold-up and specific surface area. A tracer test with lithium was used to validate this approach, and the gas mixing in the filters was studied using continuous CO2 and O2 measurements. With the tracer test results, the biodegradation model was adapted, and it became clear that biodegradation and adsorption to solids can be neglected. On this basis, a simple dynamic mass balance model was built. Simulations with this model reveal that changing the air flow rate in the trickling filter system has little effect on the VOC stripping efficiency at steady state. However, immediately after an air flow rate change, quite high flux and concentration peaks of VOCs can be expected. These phenomena are of major importance for the design of an off-gas treatment facility. PMID- 11385861 TI - Sensitivity analysis and calibration of the parameters of ESWAT: application to the River Dender. AB - The paper deals with the sensitivity analysis and parameter calibration of a complex river water quality model, implemented in ESWAT. The Extended SWAT includes a QUALIIE-based river quality simulator, in view of an integrated analysis of water quantity and quality management practises. The sensitivity analysis uses Latin Hypercube Sampling and criteria related to the duration of low concentrations of dissolved oxygen and the occurrence of high algae concentrations. The analysis on the river Dender shows that parameters related to the growth and die-off of the algae have the largest impact, while also the BOD decay constant and the benthic oxygen demand are important. A subsequent calibration of these most important parameters shows however that the optimal values of the parameters related to the activity of the algae are statistically not significant. This apparent contradiction is due to the poor information content of the measurements. It is concluded that the application illustrates the complementarity of the sensitivity analysis and the parameter calibration. PMID- 11385862 TI - Fast, simultaneous simulation of the integrated urban wastewater system using mechanistic surrogate models. AB - The urban wastewater system components (sewer, treatment plant, and river) are often modelled using complex mechanistic models. Mechanistic surrogate models are introduced here as simplified models that still contain some physical knowledge. Surrogate models are faster, but are less but still sufficiently accurate, and require more data to be calibrated. The possibilities of replacing actual field data by virtual data generated with a complex mechanistic model for calibration of the surrogate model are examined. As an example, a series of tanks with variable volume is shown to approximate sufficiently well the flow propagation in the river Zwalm (Belgium) as predicted by the "de Saint-Venant" equations. The three surrogate models can be implemented in the WEST simulator, which makes a simultaneous simulation of the system possible. In this work a connection is made between the ASM1 and the new IWA River Model No. 1 (RWOM1) by using a translator between the models in such a way that both mass and elemental balances remain closed for the overall system. This approach is illustrated with a case study on the river Lambro (Italy). The dispersion process in this river with steady flow could be modelled by using a tanks in series model, while the water quality in the river was predicted to improve substantially with an increase in hydraulic capacity of the treatment plant. The simulation results with the upgraded plant still need to be checked by field data. PMID- 11385863 TI - Ranking stormwater control strategies under uncertainty: the River Cam case study. AB - Monte Carlo simulations taking uncertainty in model parameters into account were performed on a river water quality model. The simulation results were used to rank wastewater treatment plant control strategies according to their impacts on river water quality. This impact is estimated by the maximum ammonium concentration and by the duration of dissolved oxygen concentration below 4 g/m3 at the downstream boundary of the system. The strategies were classified according to the previous criteria using 4 ranking methods, one of them being based on the concept of stochastic dominance. Results are presented for a case study based on a 10 km stretch of the River Cam as it passes through the city of Cambridge in Eastern England. It was found that ranking was robust in face of uncertainty in the parameter values for the control strategies considered as being superior in terms of river water quality impacts. PMID- 11385864 TI - Integral water quality modelling of catchments. AB - ESWAT--Extended Soil and Water Assessment Tool--was developed to allow for an integral modelling of the water quantity and quality processes in river basins. The diffuse pollution sources are assessed by considering crop and soil processes and--together with the point sources--further transformed by an in-stream water quality module. An autocalibration procedure allows for the optimisation of the process parameters. The optimisation uses a global optimisation criterion, whereby several objective functions can be considered or combined and whereby several output variables can be taken into account simultaneously. The model and the calibration procedure are applied to the river Dender in Belgium. PMID- 11385865 TI - Identifiability and uncertainty analysis of the river water quality model no. 1 (RWQM1). AB - State of the art models as used in activated sludge modelling and recently proposed for river water quality modelling integrate the knowledge in a certain field. If applied to data from a specific site, such models are nearly always overparameterised. This raises the question of how many parameters can be fitted in a given context and how to find identifiable parameter subsets given the experimental layout. This problem is addressed for the kinetic parameters of a simplified version of the recently published river water quality model no. 1 (RWQM1). The selection of practically identifiable parameter subsets is discussed for typical boundary conditions as a function of the measurement layout. Two methods for identifiable subset selection were applied and lead to nearly the same results. Assuming upstream and downstream measurements of dissolved substances to be available, only a few (5-8) model parameters appear to be identifiable. Extensive measurement campaigns with dedicated experiments seem to be required for successful calibration of RWQM1. The estimated prior uncertainties of the model parameters are used to estimate the uncertainty of model predictions. Finally an estimate is provided for the maximum possible decrease in prediction uncertainty achievable by a perfect determination of the values of the identifiable model parameters. PMID- 11385866 TI - A numerical identifiability test for state-space models--application to optimal experimental design. AB - This paper describes a mathematical tool for identifiability analysis, easily applicable to high order non-linear systems modelled in state-space and implementable in simulators with a time-discrete approach. This procedure also permits a rigorous analysis of the expected estimation errors (average and maximum) in calibration experiments. The methodology is based on the recursive numerical evaluation of the information matrix during the simulation of a calibration experiment and in the setting-up of a group of information parameters based on geometric interpretations of this matrix. As an example of the utility of the proposed test, the paper presents its application to an optimal experimental design of ASM Model No. 1 calibration, in order to estimate the maximum specific growth rate microH and the concentration of heterotrophic biomass XBH. PMID- 11385867 TI - Practical identifiability of model parameters by combined respirometric titrimetric measurements. AB - An earlier study on theoretical identifiability of parameters for a two-step nitrification model showed that a unique estimation of the yield YA1 is possible with combined respirometric-titrimetric data, contrary to the case where only one type of measurement is available. Here, the practical identifiability of model parameters was investigated via evaluation of the output sensitivity functions and the corresponding Fisher Information Matrix (FIM). It appeared that the FIM was not sufficiently powerful to predict the practical identifiability of this case with combined measurements as parameters could indeed be identified despite the fact that the FIM became singular. The accuracy of parameter estimates based on respirometric and titrimetric data and combination thereof was also investigated. Estimation on titrimetric data (Hp) was very accurate and a fast convergence of the objective function towards a minimum was obtained. The latter also holds for estimation on oxygen uptake rate data (rO), however with a lower accuracy. Parameter estimation based on oxygen concentration data (SO) was more complex but resulted in a higher accuracy. Thus, when the highest accuracy is needed it is recommended to estimate parameters initially on Hp and/or rO data, and to subsequently use these parameters as initial values for final, and more accurate estimation on SO data. PMID- 11385868 TI - Parameter estimation procedure for complex non-linear systems: calibration of ASM No. 1 for N-removal in a full-scale oxidation ditch. AB - When applied to large simulation models, the process of parameter estimation is also called calibration. Calibration of complex non-linear systems, such as activated sludge plants, is often not an easy task. On the one hand, manual calibration of such complex systems is usually time-consuming, and its results are often not reproducible. On the other hand, conventional automatic calibration methods are not always straightforward and often hampered by local minima problems. In this paper a new straightforward and automatic procedure, which is based on the response surface method (RSM) for selecting the best identifiable parameters, is proposed. In RSM, the process response (output) is related to the levels of the input variables in terms of a first- or second-order regression model. Usually, RSM is used to relate measured process output quantities to process conditions. However, in this paper RSM is used for selecting the dominant parameters, by evaluating parameters sensitivity in a predefined region. Good results obtained in calibration of ASM No. 1 for N-removal in a full-scale oxidation ditch proved that the proposed procedure is successful and reliable. PMID- 11385869 TI - A systematic approach to error isolation in computerized wastewater simulation models. AB - Activated sludge models are used extensively in the study of wastewater treatment processes. While various commercial implementations of these models are available, there are many people who need to code models themselves using the simulation packages available to them. Quality assurance of such models is difficult. While benchmarking problems have been developed and are available, the comparison of simulation data with that of commercial models leads only to the detection, not the isolation of errors. To identify the errors in the code is time-consuming. In this paper, we address the problem by developing a systematic and largely automated approach to the isolation of coding errors. There are three steps: firstly, possible errors are classified according to their place in the model structure and a feature matrix is established for each class of errors. Secondly, an observer is designed to generate residuals, such that each class of errors imposes a subspace, spanned by its feature matrix, on the residuals. Finally, localising the residuals in a subspace isolates coding errors. The algorithm proved capable of rapidly and reliably isolating a variety of single and simultaneous errors in a case study using the ASM1 activated sludge model. In this paper a newly coded model was verified against a known implementation. The method is also applicable to simultaneous verification of any two independent implementations, hence is useful in commercial model development. PMID- 11385870 TI - An overview of the posters presented at Watermatex 2000. I. New models/integrated urban wastewater systems/time series analysis. AB - This paper presents an overview of the posters presented in the sessions 1, 6 and 9 of the Watermatex 2000 conference. The first session focused on the development of new models in different areas of environmental technology, e.g. wastewater, ground pollution, sewers, etc. The sixth session dealt with integrated urban wastewater systems. Session 9 focused on the application of neural network modeling and principal component analysis in time series analysis. Rewarded posters are mentioned and selected for full paper publication in this issue of Wat. Sci. Tech. PMID- 11385871 TI - An overview of the posters presented at Watermatex 2000. II. Sensor/monitoring, control and decision support systems. AB - This paper gives an overview of the poster sessions on sensor/monitoring, control and decision support systems, as they have been presented during the Watermatex 2000 conference. The COST benchmark for the comparison of different control strategies has been analysed. Two different teams have performed research on the control of sewers to decrease combined sewer overflows. Extended methods for gauge monitoring accuracy are proposed. The use of adaptive controllers for improving control performance under varying process conditions is demonstrated in two applications. More advanced techniques for monitoring and control are also discussed. Applications are situated in the field of fault detection and control. Three posters on decision support systems have presented the methodology and architecture of specific applications. Rewarded posters are mentioned and have been selected for full paper publication in this issue of Wat. Sci. Tech. PMID- 11385872 TI - An overview of the posters presented at Watermatex 2000. III: Model selection and calibration/optimal experimental design. AB - This paper presents an overview of the posters presented in sessions 7 and 8 of the Watermatex 2000 conference. These posters present two aspects of modelling biological processes--model selection and calibration. Special attention is given to the papers on OED (Optimal Experimental Design), which is a method of optimising the data collection for model selection and calibration. The presence of these presentations at the conference highlights the continuing significance of modelling and stresses the requirement of improvements in modelling techniques. The papers provide some contribution to this end. PMID- 11385873 TI - Modelling and simulation of the steady-state of secondary settlers in wastewater treatment plants. AB - This paper discusses the steady-state modelling of thickening in circular secondary settlers of activated sludge processes. The limitations of the solid flux theory basic models to represent steady-state operating conditions serve as a basis to introduce more sophisticated models derived from computational fluid dynamics. Parameter identification and sensitivity studies have been performed from lab-scale continuous experiments. PMID- 11385874 TI - Development and evaluation of a mathematical model for the study of sediment related water quality issues. AB - A mathematical model (Sediment-Transport-Associated Nutrient Dynamics-STAND) has been developed for the study of sediment-associated water quality issues. The model is intended to simulate changes of water composition associated with sediment behavior. It has a 3-level structure. The first level accounts for the hydraulics of open-channel flow. The second computes sediment transport potential and actual rates based on the information provided by the first level. A non equilibrium approach is used. In the third level, changes of nutrient concentrations along a studied river are computed with the consideration of nutrient transport, adsorption/desorption, and release. In order to calibrate the model, field data were collected from the Oconee River, a major tributary of the Altamaha River in Georgia, USA. Two stations, approximately 17 km distant from each other, were established along the river for the purpose of data collection. Observations of the river's hydraulics, suspended sediment, and water quality (mainly orthophosphate, nitrate, temperature, specific conductivity, oxidation reduction potential, dissolved oxygen, and pH) were collected at the two stations. Another data set collected along a major tributary of the Yellow River in China was also used for calibration of the model's hydraulics and sediment transport parts. Calibration and validation results are encouraging, which suggests STAND may be a useful tool for the thorough study and understanding of nutrient dynamics associated with sediment behaviour. PMID- 11385875 TI - LANDFLOW: a 3D finite volume model of combined free and porous flow of water in contaminated land sites. AB - Free and porous flow of water in lands often occurs in combination under different physical circumstances. To model such configurations, open and porous flow regions need to be studied independently and also to integrate them through well-posed mathematical formulations. This paper presents a computational investigation (modelling and simulation) on flow of water and contaminants' mobility in soil. The governing partial differential equations of the flow were discretised and reduced to algebraic forms by finite volume method for simulation purposes. Dirichlet boundary conditions were used at the inlet. For sidewalls, no slip conditions were imposed while for the outer boundary, stress free boundary conditions were used. PMID- 11385876 TI - Evaluation of pH inhibition effect on activated sludge by the pseudo toxic concentration (CPT) concept model. AB - It is generally accepted that the inhibition effect of pH on activated sludge follows the non-competitive inhibition kinetics. But the non-competitive inhibition kinetic equation cannot be directly applied to pH inhibition, due to the difficulty in quantification of pH as a term of inhibitor concentration. So, many empirical equations were developed especially for acidic condition to describe pH inhibition effect. In this research, the pseudo toxic concentration (CPT) concept model to quantify pH inhibition effect on activated sludge was proposed and compared with other existed models. Prediction of performance, presented by Prediction Accuracy and Prediction Accuracy Index, showed that the CPT concept model can explain the reduction of the maximum specific growth rate (mu max) more accurately than any other models do at a wide range of pH. The CPT concept model was applicable not only to activated sludge but also to specific microorganism groups, such as nitrifier, less acidophilic species and nitrifying biofilm. PMID- 11385877 TI - Modeling of a reactive primary clarifier. AB - Even though many models have been proposed for primary clarification, none is directly compatible with the ASM1. The objective of this paper is to present the development of a reactive primary clarifier model to be used in a wastewater treatment plant simulator (WEST). A model simulating COD behavior has been developed based on the Takacs model, and was tested with full-scale data. Particulate effluent COD was well described but problems occurred predicting the underflow suspended solids concentration. The model had to be upgraded with a residence time and a flocculation term to simulate the behavior of soluble COD. An ammonification term was added to the model, resulting in an improved model fit on effluent ammonium. PMID- 11385878 TI - Long-term pollution simulation in combined sewer networks. AB - This paper presents results of long term pollution simulations on the example of the sewerage system of Grand-Couronne. This modelling work is part of a study where objective is to develop a method to define the reference flow of a WWTP. The model HYDROWORKS DM has been successfully validated in hydraulics and pollution for the sewer network, for long time simulations. A conceptual model has been built to model the pollution in the tank at the outlet of the combined system. One synthetic year of rain has been used to simulate the working of the "up stream system" of the WWTP (combined sewer + tank + separate sewer + pre treatments) and has been successfully validated by measurements of the 1998-1999 year. If this paper is focused on the "up stream system", the SIMBA/SIMBAD WWTP model has been successfully calibrated and validated too, and the combination represents a fully validated "Integrated Model" for the sewerage system. PMID- 11385879 TI - Modelling of activated sludge acclimisation to a non-ionic surfactant. AB - A model is proposed to describe activated sludge acclimatisation to a non-ionic surfactant. The model was calibrated automatically, using WEST, a specific software environment for wastewater treatment model building, simulation and parameter estimation. The assays have been performed in a sequencing-batch reactor (SBR), using a non-ionic surfactant as sole carbon source and non acclimatised sludge. The best fitting model was based on the assumption of three sequentially degraded COD fractions, where the second fraction is a metabolite of the original molecule and the third fraction is a more slowly biodegradable metabolite resulting from the secondary degradation. For primary degradation, hydrolysis with no associated growth was assumed. The growth of microorganisms responsible for degradation of the second and third COD fractions was presumed to follow Haldane and first order kinetics, respectively. The model was able to fit four consecutive assays of the same acclimatisation process, using Brij 30 as carbon source, with different food/microorganism ratios. The parameters obtained showed that the (self-)inhibition of the growth on the second COD fraction decreased along acclimatisation. PMID- 11385880 TI - Automated monitoring of activated sludge in a pilot plant using image analysis. AB - An automated procedure for the characterisation by image analysis of the morphology of activated sludge has been used to monitor the biomass in a pilot wastewater treatment plant, in complement to the usual settleability (sludge volume index, settling velocity) and size distribution (by laser granulometry) measurements. PMID- 11385881 TI - On-line viable biomass measurement and estimation of the specific growth rate of activated sludge from municipal wastewater treatment. AB - In order to control wastewater processes, on-line measurements of important process variables are crucial. This contribution focuses on the applicability of the Biomass Monitor for on-line viable biomass measurement of activated sludge from a municipal wastewater plant. In addition, the specific growth rate of the sludge is estimated on-line, based on the information derived from the device under study. Compared to dry weight measurements, the Biomass Monitor hardware offers the advantage of a biologically more appropriate observation of the biomass by only taking into account the viable cells in the population. The optimal measurement frequency of the biomass monitoring device for the given experimental conditions has been determined. Furthermore, the capacitance readings have been correlated with off-line analyses of dry weight of the sludge during the experimental phase in which no death of cells occurred. Finally, an evaluation of the estimator of the specific growth rate including its tuning is presented. PMID- 11385882 TI - Defining myocardial infarction: not too late for common sense or consistency? PMID- 11385883 TI - The National Confidential Enquiry into Perioperative Deaths 2000: then and now. PMID- 11385884 TI - Caring for others can seriously damage your health. AB - To care for others is a privilege and a source of personal and job satisfaction. However, caregivers must also consider the implications of their work for their own health and welfare. PMID- 11385885 TI - Post stroke depression. AB - Depression following stroke is common. Although it is highlighted as an issue in stroke guidelines, guidance on diagnosis or management is not given. This paper presents the original research from a literature review of Medline and the Cochrane Database on stroke and depression, and discusses some of the clinical implications of the findings. PMID- 11385887 TI - Pharmacological management of opiate withdrawal. AB - Opiate misuse is extremely prevalent in the UK. A high proportion of these individuals experience withdrawal symptoms. This article describes the clinical features and discusses the current literature on the pharmacological management of opiate withdrawal. PMID- 11385886 TI - St John's wort for depression: what's the evidence? AB - There is a moderate amount of evidence to suggest that St John's wort is better than placebo in the short-term management of mild to moderate depressive illness, but there is increasing concern over reports of drug interactions. This article examines the scientific evidence and discusses relevant clinical issues. PMID- 11385888 TI - Ascitic fluid analysis: the role of biochemistry and haematology. AB - In specific settings, biochemical and haematological analysis of ascitic fluid may provide answers to important clinical questions. This review seeks to outline the value and limitations of ascitic fluid analysis and the main clinical scenarios in which it may be useful. PMID- 11385889 TI - Case 2: the interpretation of ST segment changes in the presence of bundle-branch block. PMID- 11385891 TI - The medical masterclass: a new tool for continuing professional development. AB - The Medical Masterclass comprises twelve paper-based modules, two CD-ROMs and a companion website. Its main aim is to help doctors in their first few years of training to improve their medical skills and knowledge, and in particular in their preparation for the MRCP examination. It will also be a valuable tool for continuing professional development. PMID- 11385890 TI - Systemic treatment of metastatic breast cancer. AB - This article reviews recent advances in the treatment of metastatic breast cancer, in particular the results of randomized controlled phase III clinical trials. These have led to the evidence-based introduction of several new drugs including aromatase inhibitors, taxanes and bisphosphonates, some of which have been shown to impact on overall survival. PMID- 11385892 TI - The future of journals: where will electronic publishing take us? AB - Journals provide much that is of immense value to the researcher and practitioner. Yet the economics of journals are unsustainable; the amount of publishable research is increasing more rapidly than the funds available to buy it. Various alternative models are emerging, none of them without problems. The way forward is not clear, but change is inevitable. PMID- 11385893 TI - Stress and motivation. PMID- 11385894 TI - Thinking before prescribing. PMID- 11385895 TI - Pulmonary embolism and the electrocardiogram. PMID- 11385896 TI - Non-steroidal hip arthropathy: case histories and differential diagnosis. PMID- 11385897 TI - To transfuse or not to transfuse: iatrogenic compromise of women's reproductive careers. PMID- 11385898 TI - A patient with hypertrophic obstructive cardiomyopathy presenting for total knee replacement. PMID- 11385899 TI - [Can one reduce the morbidity related to vascular access for hemodialysis?]. PMID- 11385900 TI - [Stent migration in a hemodialysis patient]. AB - Haemodialysis is the most frequently used treatment of chronic renal failure which cannot be performed with out vascular access, of which arteriovenous fistula is the most frequently used. One of the complication of arteriovenous fistula is venous stenosis. Its treatment is usually percutaneous angioplasty associated or not with stent. Rare cases of migration of stent have been reported. PMID- 11385901 TI - [Pulmonary migration of an axillary stent in a chronic hemodialysis patient]. AB - The short-lived results of angioplasty for venous stenosis of a dialysis access led to stent placement. Its migration is a rare but dangerous complication, because of its final destination, that is right heart or pulmonary artery. We report on such a case and compare it with three cases found in literature. PMID- 11385902 TI - [Rosai-Dorfman disease revealed by renal failure: case report]. AB - We report a case of Rosai-Dorfman Disease revealed by renal failure in a 43 years old patient. Clinical presentation included abdominal lymphadenopathy and general status deterioration. Diagnosis was established by histopathological examination of the node which revealed sinusal lymphohistiocytosis. Treatment combined prednisone and cyclophosphamide and was effective with regression of renal failure. We will review the diagnostic criteria and the prognosis of this disorder of unknown etiology. PMID- 11385903 TI - [A possible adverse effect from the association of losartan-mefenamic acid in hemodialysis]. PMID- 11385904 TI - [Psychosomatic gynecology and obstetrics]. PMID- 11385905 TI - [Chronic pelvic pain syndrome--a multifactorial syndrome]. AB - OBJECTIVE: As a clinically significant syndrome in the field of gynaecology, chronic pelvic pain syndrome (CPPS) leads to illnesses which are prolonged over many years and, in some cases, to a high incidence of surgical intervention. Answers to questions about its aetiology, gynaecological associations, such as adhesions or endometriosis, and new approaches to diagnosis and treatment are urgently required. PATIENTS AND METHODS: Psychometric data were evaluated in 106 patients with CPPS and in 36 clinical controls with vulvodynia. Psychological testing was undertaken in 68 patients with CPPS and 24 with vulvodynia, in comparison with a control group of healthy women (n = 34). DSM IV and ICD 10 classifications were used and neurosis-orientated diagnoses were also assigned. In order to investigate musculoskeletal diseases in CPPS, these patients (106/36) underwent orthopaedic examination and were compared with a control group with no pain (n = 35). RESULTS: An interdisciplinary diagnostic work-up proved to be of clinical relevance, with a significantly higher frequency of cases of facet syndrome in CPPS. After an interdisciplinary diagnostic work-up and treatment directed towards psychosomatic symptoms, the gynaecological differential diagnosis may possibly include laparoscopy. The psychological aggression conflict associated with a significantly greater number of patients with so-called "early disturbance" and latent depression can find pronounced expression during treatment. Results of qualitative investigations in patients with endometriosis show typical psychological symptoms. CONCLUSIONS: Treatment that is both interdisciplinary and strictly orientated towards psychosomatic disorders seems appropriate to this complex syndrome. In particular, the psychological dynamics of an aggression conflict in the patient must be taken into consideration, so that this conflict does not transmit to the treatment. PMID- 11385906 TI - [Integrated gynecology: new medical treatments due to psychosomatic methods of evaluation]. AB - OBJECTIVE: Women with gynecologic cancer are confronted with difficult decisions regarding the therapeutic options. The objective of the present paper is to demonstrate the relationship between surgical procedures and the outcome on quality of life and to discuss the implications for patient management. METHODS: Gynecologic patients were assessed in a prospective study with preoperative semistructured interviews and objective assessments (T1), interviews were repeated 4 and 12 months postoperatively (T2, T3). RESULTS: Women planned for hysterectomy with severe complaints indicate a better postoperative quality of life. Cancer patients, however, tend to feel more distressed about the surgical procedure if they could not be treated organ preserving or by reconstructive techniques. Medical interaction is dependent on the patient's anxiety level and mostly important for their quality of life before and after surgery. CONCLUSION: Psychosomatic research is not only necessary to understand the patient's needs before and after surgery but may also serve as an evaluation method of therapeutical options. By this methods we are able to anticipate the medical and psychological consequences of the therapeutic decisions. Future studies will systematically explore the alternating effect of surgical procedures on the patient's well-being. PMID- 11385907 TI - [Body image in gynecologic patients before and after radical surgery]. AB - After radical gynecologic surgery women are faced with therapy-induced changes of their body. Since 1995, the body images of women who undergo pelvic exenteration, Wertheim-Meigs-operation or hysterectomy are assessed preoperatively and four and twelve months postoperatively. The aim of this multidimensional prospective study is to get basic information for effective counselling and support. One year after hysterectomy women state to have a normal body image. Cancer patients feel less attractive, less self-confident and more discontented in sexuality depending on therapy-induced changes of their bodies. These women could profit from problem related preoperative counselling. Postoperative counselling offers will be helpful to support women in the process of acceptance of bodily changes and in the reorganisation of their sexual life. PMID- 11385908 TI - [Consenting and declining patients for an intervention group after breast cancer surgery differ in terms of quality of life, coping and immunological functional assays]. AB - OBJECTIVE: Psychosocial intervention during the course of cancer is still being controversially disputed in psychosomatic studies. Immunological and endocrinological parameters can serve as links in observing the influence of the interactive processes involved. MATERIAL AND METHODS: The approach to determine whether psychotherapeutic intervention can lead to an improved quality of life on the one hand, and to a change in immune defenses in breast cancer patients as well, employed using the EORTC QLQ-C30 and optimistic self-efficacy (OKE) as well as evaluating lymphocyte subpopulations, lymphocyte proliferation and NK cell activity. Fifty-three breast cancer patients were offered the chance to participate in a psychosocial intervention group. Twenty-three accepted the offer. Randomized sampling of the group participants was determined twice by using the 30 EORTC QLQ-C30 parameters. In addition, immune function was measured before beginning (T1) and after completing the group intervention sessions (T2). This data was compared with that of the declining group. RESULTS: The analysis of the individual EORTC parameters in the pre- and post-comparison showed a decrease in "emotional strain" and an increase in "all-around quality of life" at the T2 test period. NK cell activity was increased at T2 as well in the participating group and was higher than that of the declining group both at T1 and T2 testing times. CONCLUSION: Participation in the psychosocial intervention group resulted in a marked decrease in the emotional strain and an increase in the quality of life. The increased NK-cell activity with lowered T-cell activation in comparison to the decliners hints at a coherence between coping behaviour and changes of the immunological cell function. The results of this study will be investigated further in a greater sample. PMID- 11385909 TI - [Female menopause--comparison of knowledge and opinion in German and Turkish-born women]. AB - 320 German and 262 Turkish immigrant patients of the gynaecological units of the Virchow Women's Clinic in Berlin were questioned, to compare their knowledge and opinion on menopause and possible cultural influence factors. The results showed that 70% of the migrant vs. 90% of the German women regarded menopause as a normal phase in a women's life and more Turkish than German women (35% vs. 7%) associated complaints with menopause. However the immigrant women knew noticeably less about health risks related to menopause (10% vs. 50%). PMID- 11385910 TI - [Termination of pregnancy for fetal abnormality--a traumatic experience?]. AB - The diagnosis of a lethal anomaly of the fetus can affect a pregnant woman in a traumatic way. Almost immediately she has to decide whether she wishes the pregnancy to be terminated or not. Literature shows that such a loss is very difficult to cope with, and can lead to social isolation and depression. Contrary to popular belief the loss felt by the woman is at least the same to that following a stillbirth. Problems arise when the woman has difficulties in expressing her feelings, has a lack of self-esteem or receives very little social support. The prenatal diagnosis evokes an acute grief reaction. Only few studies are available regarding length, course and severity of grief in this case. Although an abortion through a fetal anomaly is a traumatic experience, research is vague on the trauma caused. Present day research of the psychological sequelae after the termination will be summarized. In respect to the current trauma research lies the question of which psychiatric conditions arise from such a traumatic experience. PMID- 11385911 TI - [Psychologic effects of traumatic live deliveries]. AB - OBJECTIVES: Traumatically experienced childbirth can lead to serious psychological disturbances postpartum. Dependent upon objective and subjective factors, some women may even develop the symptomatology of a posttraumatic stress disorder (PTSD). The aim of a pilot study was the evaluation of the frequency of traumatically experienced childbirth, of the effects of these traumatic experiences and possible risk factors for the development of PTSD. STUDY DESIGN: 976 women, who had given birth at the Bonn University Women's Hospital during 1997/1998, were retrospectively questioned regarding their experiences with pregnancy, childbirth and postpartum. 46 women, who described relevant psychological symptoms after childbirth, were personally interviewed. RESULTS: 424 women returned the completed questionnaire. 17.2% of the women reported anxiety postpartum, 9.4% depressive symptoms, 12% a mental re-experience of delivery within the first weeks postpartum, 3.8% were still suffering from these intrusions at the time of the study. In the first weeks after labor, nightmares were reported in 3.1%. In 4 cases, the full criteria for a PTSD were met. In 10 further cases, a subsyndromal form of this disorder was found. The case analysis showed that the development of PTSD symptoms was influenced by factors such as expectations, need for control, sense of shame and previous traumatic experiences. CONCLUSIONS: Psychological symptoms postpartum were reported frequently. Traumatically experienced childbirth can be responsible for specific short-term or long-term symptoms. In individual cases, a PTSD can develop after a traumatic delivery with long-term negative consequences for the health and mental condition of the mother, the mother-child-relationship and the desire for further pregnancy. In such cases, a specific psychotherapeutic treatment is always necessary. PMID- 11385912 TI - [Specific concepts of care for pregnant women with drug addiction or HIV infection]. AB - In the last 10 years about 130 women with a drug addiction and more than 100 HIV positive pregnant women were treated at the 1. University Hospital of Obstetrics and Gynaecology in Munich. Besides a specialized medical treatment both groups required intensive psychosocial care. HIV-infected people are still isolated and suffer from the social stigmata. Their essential needs for sexuality and children of their own are often ignored or even condemned because of irrational fears about HIV, which continue despite rapid medical improvements. The life-expectancy for example has increased since the inauguration of protease inhibitors. Vertical transmission of HIV is below 2% through medical treatment in pregnancy, elective cesarean section and renunciation of breastfeeding. Drug addicted pregnant women are given the opportunity to change their life in order to care for their children appropriately. The basis for this is a substitution with levomethadone and elimination of the use of other drugs. The addicted women often can reduce the dosage of levomethadone during the course of their pregnancy and sometimes can cease totally. Normally they are highly motivated and thus can ease the withdrawal symptoms of their newborns following delivery. By establishing a reliable social net during pregnancy mothers learn to recognize the demands of their children after birth and thus emotional and cognitive deficits can be prevented. PMID- 11385913 TI - [Cesarean section on request--a medical and psychosomatic problem]. AB - Caesarean section on request of the mother turned out lately to be very actual in obstetrical discussions. To come to rational decisions the scientific results of several medical disciplines should be taken into consideration. There is no scientific evidence for benefits of such elective c-section in paediatrics and psychosocial medicine. The obstetricians view shows both benefits and disadvantages, the later overweighing the first. In the light of these data, elective caesarean without other indication but the mothers request does not show to be an acceptable routine procedure. PMID- 11385914 TI - [Child or career? Desire for a child in East and West Berlin]. AB - OBJECTIVE: In the Federal Republic of Germany the rate of the childless women rises. While the topic of unwanted childlessness among gynecologists receives broad attention among gynecologists, the question of (temporaryly?) intended renouncement of pregnancy and birth is of little interest. For the former GDR a low age with the firstborn and a high mother rate were typically. After the "turn" serious modifications of the birth rate in East Germany happened. MATERIAL AND METHODS: The available study examined 1996 motives for and against a child with 554 up-to-date wanted childless women in East- and West-Berlin (population referred sample). In the questionnaire, besides sociodemographic data, information was raised to own childhood, partnership, occupation, satisfaction with life and for child desire motivation. RESULTS: Apart from the school and vocational termination conceptions on the age with the firstborn, disadvantages for a life with child and value hierarchies for partnership and family were important for the description of the dilemma of compatibility of occupation and family. Eastsocialized women consider a compatibility rather possible. CONCLUSIONS: Targets for vocational development and responsible parenthood for young women come into conflict. Temporal delaying of the child desire leads to an increase of intended and unwanted childlessness with various (psychosomatic) effects on obstetrics and gynecology. PMID- 11385915 TI - [Children--really why not... Results of a representative survey in East and West Germany]. AB - The topic "wish to get a child" is of interest in scientific research for several reasons. One reason is the dramatic drop in the birth rate ibn Eastern Germany following reunification between 1990-1995, one other reason is the declining birth rate as a constant fact in Germany as a whole. Resulting from this demographic phenomenon the question was to detect the determinants of wish to get a child and their positive or negative influences regarding the realization. To answer this question we developed an theoretical model which focussed the decision process. This model was tested in a representative survey in 1999. This article presents some first results of the study. The main finding was that the wish to get a child is very strongly influenced by emotional aspects. PMID- 11385916 TI - [Etiology and coping with involuntary childlessness--are there specificic motivations for wanting a child?]. AB - OBJECTIVE: Do patients with functional sterility have special motivations to have a child? Could these motivations be an underlying reason for their sterility opposed to patients with organic sterility? Are there any specific motivations to have a child in patients with strong desires to have children? MATERIAL AND METHODS: 80 patients attending an infertility clinic were evaluated using an interview, questionnaire and Giessen-Test (GT). RESULTS: The motivation to have a child is not different in women with organic and functional sterility. Women with a strong desire to have children express significantly less often, that a child would contribute for them to achieve fulfilling life. But they express more frequently the hope to find a successful treatment for their sterility. CONCLUSIONS: It can not be conducted that psychosomatic disorders are the true cause for sterility in absence of organic pathology. This would not consider the complexity of psychosomatic processes. Coping mechanism are more important than the underlying etiology of sterility. PMID- 11385917 TI - [Is there an effect of psychosocial factors on pregnancy rates in sterility therapy?]. AB - OBJECTIVE: We examined the possible connection between mood state of patients during IVF/ICSI-treatment and pregnancy rate. Which influence does the patient's age and the social support have on the mood state? MATERIAL AND METHODS: Questionnaires were handed out to 100 patients of our fertility center during an IVF/ICSI-treatment-cycle. We used the standardized Leipziger Stimmungsfragebogen (LSB), a visual analogue-scale for free indication of mood state and the questionnaire for social support (SOZU). RESULTS: Per oocyte pick up 32 women became pregnant (32.7%), 58 women did not become pregnant (59.2%). Lower rating on the aggressivity-scale showed a tendency (p = 0.085) and higher rating on the balance-scale showed a trend toward higher pregnancy rates. There was no significant relation shown on the visual analogue-scale as well as on the LSB scales grief, activity and fatigue. There was no correlation between age and mood state or between social support and pregnancy rate. CONCLUSIONS: We could not show a significant relation between general mood state and pregnancy rate. Yet there was a tendency concerning certain components of mood. Increasing age of the patient does not have a negative influence on the mood state either. The individual situation and adjustment seem to be decisive. PMID- 11385918 TI - Function of the posterior tibial tendon muscle. AB - The posterior tibial tendon muscle plays a critical role in the function of the foot. It is, with the exception of the triceps surae, the largest of the extrinsic foot muscles. It is called upon to provide a critical posturing and stabilizing function to the foot during gait. This review discusses the anatomy, physiology, functional characteristics, and measurement of the strength of this muscle. PMID- 11385919 TI - Subtalar arthrodesis for treatment of posterior tibial tendon insufficiency. AB - Subtalar arthrodesis is an effective treatment of the planovalgus deformity of posterior tibial tendon insufficiency that provides stable and reliable results with minimal complications. Disadvantages include the risk of symptomatic adjacent joint arthrosis at long-term follow-up and less clinical and radiographic correction of the deformity as compared with other reconstructive options, which may make the joint-preserving procedures more attractive for the primary treatment of patients with a flexible pes planovalgus deformity without subtalar pain. PMID- 11385920 TI - The adult acquired flatfoot and spring ligament complex. Pathology and implications for treatment. AB - This article reviews the anatomy of the spring ligament complex, the extent of ligament involvement in addition to the spring ligament, and implications for clinical reconstruction. PMID- 11385921 TI - Posterior tibial tendon insufficiency. Isolated fusion of the talonavicular joint. AB - In selected patients, fusion of the talonavicular joint can be an effective treatment of adult flatfoot deformity. Restriction of motion and altered hindfoot mechanics, however, are a consequence of talonavicular fusion and can lead to accelerated arthrosis of adjacent joints. In patients with severe long-standing deformity, medial displacement calcaneal osteotomy may be a necessary adjunct to talonavicular fusion for adequate correction of heel valgus. PMID- 11385922 TI - Biomechanics of procedures used in adult flatfoot deformity. AB - A flatfoot deformity can occur secondary to fairly obvious causes, or more subtle and less definable entities. Complicating the situation further, it is likely that the cause of an acquired flatfoot deformity in an adult is multifactorial. This likelihood makes the definition, diagnosis, and appropriate treatment of this condition a daunting task. More research is needed to define further the biomechanics of the foot and to understand the significance of the forces that combine to create flatfoot deformity. PMID- 11385923 TI - Treatment of stage 3 adult acquired flatfoot. AB - Stage 3 adult acquired flatfoot occurs when chronic posterior tibial tendon insufficiency results in fixed hindfoot valgus or fixed forefoot abduction and supination. Nonoperative management results in limited success. Corrective fusion is the treatment of choice. Although a variety of arthrodeses have been employed, triple arthrodesis remains the gold standard. PMID- 11385924 TI - Treatment of stage 4 adult acquired flatfoot. AB - Stage 4 PTT dysfunction is a rare anatomic condition in which fixed hindfoot valgus is associated with valgus tilting of the talus within the ankle mortise. Success with nonoperative management is the exception rather than the rule. The surgical options are a tibiotalocalcaneal fusion or a pantalar fusion; however, there are few results reported in the adult acquired flatfoot population. Valgus talar tilting after triple arthrodesis may be the challenge of the future. PMID- 11385925 TI - Complications of surgical treatments for adult flatfoot deformities. AB - Numerous surgical procedures have been proposed for reconstructing and correcting adult flatfoot deformity. Most procedures include lengthening of the tight Achilles tendon and transferring the FDL tendon to substitute for a dysfunctional PTT. These procedures commonly are combined with a bone procedure to stabilize the correction and correct the pathologic anatomy. All of these procedures have the potential for a range of complications, many of them serious. In general, the clinician should perform the least invasive procedure that decreases pain and improves function. The effects of each procedure and the associated morbidity and complications must be considered. PMID- 11385926 TI - The use of orthotic devices in adult acquired flatfoot deformity. AB - PTT dysfunction is the most common cause of adult acquired flat foot deformity. The aggressive nonoperative approach has become accepted more widely, in part because of the advances in orthotic and bracing technology and options. Many patients with a PTT dysfunction can be treated effectively with conservative management protocols. The goal of alleviating pain and correcting deformities is being accomplished with the proper application of the wide spectrum of orthotic modalities available today. PMID- 11385927 TI - Tenosynovitis of the posterior tibial tendon. AB - PTT tenosynovitis is a recognized entity no longer confused with an ankle sprain. Three possible causes are (1) overuse or age related (mechanical in cause, true stage I disease), (2) seronegative spondyloarthropathies (clinical suspicion, hematologic analysis), and (3) rheumatoid arthritis (deformity may be owing to ligamentous or capsular destruction). The PTT has a hypovascular zone 40 mm proximal to the insertion of the tendon and 14 mm in length. Pain often is localized to this portion of the tendon (primarily in stage I disease). Ultrasound is an inexpensive and accurate method to assist in the diagnosis of this condition and may replace MR imaging as more experienced ultrasonographers appear. The initial management of PTT tenosynovitis includes tendon rest and nonsteroidal anti-inflammatory medication and physical therapy. Surgical synovial debridement is performed early (6 weeks) in patients with enthesopathies (seronegative disease). This procedure may be delayed 3 months in patients with true stage I disease. At surgery, the undersurface of the tendon must be inspected for longitudinal split tears, and these must be repaired with nonabsorbable suture, burying the knots. The excursion of the tendon should be checked intraoperatively. Patients with stage I disease should be evaluated carefully for preoperative structural deformity to choose the appropriate surgical procedure and prevent failure of isolated tenosynovectomy. PMID- 11385928 TI - Flexor digitorum longus transfer with medial displacement calcaneal osteotomy. Biomechanical rationale. AB - The rationale behind combining a medical displacement calcaneal osteotomy with a flexor digitorum longus (FDL) transfer is to realign the valgus heel under the mechanical axis of the leg and to reduce the deforming valgus moment of the gastrocnemius soleus muscle group on the hindfoot. This reduces the antagonistic force on the relatively weak FDL transfer. This combination has a potential for producing a more mechanically balanced foot and acts as a double tendon transfer in which the Achilles tendon is transferred medially in addition to the FDL transfer, which substitutes for the degenerative posterior tibial tendon. PMID- 11385929 TI - Posterior tibial tendon dysfunction. Treatment by flexor digitorum longus transfer. AB - Posterior tibial tendon dysfunction presents the clinician with such a broad spectrum of clinical problems that treatment must be based on the anatomic alignment of the foot as determined by the physical examination. There is no single method to treat PTT dysfunction, but rather a variety of conservative and operative procedures that are based on the pathologic anatomy. PMID- 11385930 TI - Medial slide calcaneal osteotomy. Technique, patient selection, and results. AB - This article reviews the indications and the operative technique for the medial calcaneal slide osteotomy for the treatment of posterior tibial insufficiency. Patient selection, expected results, and complications of this technique are discussed. When used in combination with flexor digitorum longus transfer, the medial calcaneal slide osteotomy is an effective method of treatment for the adult acquired flatfoot associated with insufficiency of the posterior tibial tendon. PMID- 11385932 TI - Work and climate in traditional subsistence economies. PMID- 11385931 TI - Operative treatment of the difficult stage 2 adult acquired flatfoot deformity. AB - In the flexible pes planovalgus deformity of stage 2 posterior tibial tendon dysfunction, osteotomies appear to have a significant role in operative management by restoring more normal biomechanics, allowing tendon transfers to function successfully. The options when considering osteotomies for stage 2 disease include lateral column lengthening, medial displacement calcaneal osteotomy, and combined double osteotomy technique. The tight Achilles tendon should be lengthened as well. Lateral column lengthening has been used extensively for treatment of flexible flatfeet. It has been shown clinically and radiographically to address all 3 components of the pes planovalgus deformity present in stage 2 posterior tibial tendon dysfunction. Lateral column lengthening is used in combination with a medial soft tissue rebalancing procedure. The mechanism of action is still speculative but clearly is not owing to tensioning of the plantar fascia as previously thought. Despite the excellent correction of foot posture obtained by use of lateral column lengthening for adult acquired flatfoot, many clinicians have reservations about its use because of reported secondary increases in the calcaneocuboid joint pressures. This increase in pressure has been shown to occur experimentally, increasing the potential risk of calcaneocuboid joint arthrosis. This experimental evidence is supported by Phillips' study of the original Evans procedure, which resulted in a 65% incidence of calcaneocuboid joint arthrosis at 13-year follow-up. Mosier LaClair et al reported a 14% incidence of calcaneocuboid joint arthritis at 5 year follow-up after double osteotomy for stage 2 posterior tibial tendon dysfunction. This incidence has not been proved true in the remainder of the literature surrounding this procedure and its use for flexible flatfoot. To address the concern regarding potential calcaneocuboid arthrosis secondary to lateral column lengthening, calcaneocuboid joint distraction arthrodesis has been explored as an alternative technique. The results show good initial correction, but the follow-up is extremely limited, and one study reported loss of correction over time. Longer follow-up is needed to determine whether or not this technique would provide the lasting correction seen with the Evans procedure. Calcaneocuboid joint lengthening arthrodesis does result in some limitation of adjacent hindfoot motion. Although this limitation is significantly less compared with talonavicular and subtalar joint fusion, this procedure may result in increased local pressures and arthrosis of the midfoot or hindfoot. For the above mentioned reasons, longer follow-up studies are needed to determine whether calcaneocuboid joint distraction arthrodesis would prove to be a reliable and safe alternative for lateral column lengthening in the treatment of adult acquired flatfoot. Medial displacement calcaneal osteotomy has been used for correction of the pes planovalgus foot in posterior tibial tendon dysfunction. It has been used extensively for the surgical treatment of flexible flatfoot throughout the literature. Medial displacement osteotomy, in combination with flexor digitorum longus tendon transfer, can address all 3 components of adult acquired flatfoot. It does not recreate the medial longitudinal arch in all patients, however. Although the mechanism of action of medial displacement calcaneal osteotomy is unknown, it has been proved that it is not through the tightening of the plantar fascia in a windlass effect as previously thought. In contrast to lateral column lengthening, however, medial displacement calcaneal osteotomy does address the deforming valgus force of the Achilles tendon. Functionally transferring the insertion of the Achilles tendon medially removes a constant valgus-deforming force. The osteotomy can then act as a double tendon transfer with the flexor digitorum longus tendon to aid in foot inversion. For stage 2 posterior tibial tendon insufficiency, the authors favor the combination double osteotomy technique with a flexor digitorum longus tendon-to-medial cuneiform tendon transfer, debridement or removal of the posterior tibial tendon, and percutaneous heel cord lengthening. Early results were positive at 1.5 years after surgery with respect to maintenance of correction and functional improvement with no evidence of calcaneocuboid arthrosis. More recently, the intermediate 5-year follow-up has been assessed for this combination of procedures, and similar results were found. There was a high rate of patient satisfaction and functional improvement, and surgical correction of the flatfoot deformity was maintained and compared favorably with the contralateral normal foot. Although the intermediate follow-up found a 14% incidence of calcaneocuboid arthrosis, 50% of these patients had preoperative evidence of calcaneocuboid joint arthritis. (ABSTRACT TRUNCATED) PMID- 11385933 TI - Analysis of cognitive and motor functioning during pubertal development: a new approach. AB - We investigated cognitive-motor abilities in 303 (156 female) school children from Zagreb, Croatia, in the age span 10 to 14 years using a newly developed chronometrical reactionmeter system (CRD). The following tests were applied: CRD 311 (simple visual discrimination of signal location), CRD-324 (short-term memory actualisation), CRD-21 (simple convergent visual orientation), and CRD-11 (arithmetically conceptualised/operationalised convergent thinking). In both gender a statistically significant age related improvement of the performance for time related parameters (minimum time of test item solving (MT), total ballast (TB), and total time of test solving (TT) was observed. In contrast, the number of errors (NE), which was the only non-time related parameter tested, did not significantly change with age. Significant differences between boys and girls were observed for the time related parameters TB and MT. TB was significantly lower in girls, whereas boys tended to be faster in MT measurements. In TT as a composed measure of the mentioned parameters, no major differences were observed. We conclude that the CRD system is a new useful tool for investigating the complexity of cognitive-motor abilities in children. Our cross-sectional study demonstrated that the time-related parameters were significantly affected by age and gender during puberty. PMID- 11385934 TI - Physiological responses to water-walking in middle aged women. AB - The purpose of the present study was to examine the physiological responses to water-walking using the Flowmill, which has a treadmill at the base of a water flume, in two groups of women. In the first group, the women were known to regularly swim and exercise in water (group A), while in the second, they did not routinely participate in water-exercise (group B). In both groups, twelve healthy female volunteers in their fifties participated in the study. All of the subjects walked in water using the Flowmill for the first time. Subjects completed four consecutive bouts of 4-minute duration at progressively increasing speeds (20, 30, 40, and 50 m.min-1), with 1-minute rests between each bout. In addition, water-velocity was adjusted to the walking speed of each bout. The water-depth of the Flowmill was the level of the xiphoid process. The water and room temperatures were 30.3 +/- 0.1 degrees C and 24.9 +/- 0.4 degrees C, respectively. In both groups, the relationship between walking speed and oxygen uptake (VO2) as well as that between walking speed and heart rate (HR) changed exponentially as the walking speed increased, and the relationship between HR and VO2 was linear. The relationship between HR and VO2 was similar in both groups, and there was no significant difference between the predicted maximal oxygen uptake (VO2max) of the two groups. VO2 and HR of group B during water-walking, however, were significantly higher than those of group A at all walking speeds. The results of this study clearly showed that experience in moving through the water strongly affects physiological responses to water-exercise, even when fitness levels are equivalent. PMID- 11385935 TI - Effects of ovariectomy on intramuscular energy metabolism in young rats: how does sports-related-amenorrhea affect muscles of young female athletes? AB - The purpose of this study was to evaluate the effects of ovariectomy on intramuscular energy metabolism in young rats. Twenty-four Sprague-Dawley rats (7 weeks old) were used. Twelve of them underwent ovariectomy (OVX), and the others were sham-operated on. Seven OVX rats were examined 1-week after surgery (OVX-1 group), and the other five, 4 weeks after surgery (OVX-4 group). The gastrocnemius-plantaris-soleus (GPS) muscles group was subjected to the following measurements, and the data were compared with those of the sham group (Sham-1: n = 7, or Sham-4 group: n = 5). From the 31P-MR spectra of the GPS muscles group at rest and during electric stimulation, the muscular oxidative capacity was measured. Maximum tension and wet weight of the whole GPS muscles group were also measured. Body weight in the OVX-4 group was significantly (p < 0.01) larger than that in the Sham-4 group. The weights of the whole GPS muscles group in the Sham 1, Sham-4, OVX-1 and OVX-4 groups were 1.17, 1.51, 1.25 and 1.71 (g), respectively. The muscle weight in the OVX group tended to be greater than that in the Sham group (p < 0.10). The maximum tension and oxidative capacity did not differ significantly among the groups. These data indicated that in young rats, ovariectomy induced an increase in body and muscle weight, but did not affect the maximum tension nor oxidative capacity. PMID- 11385936 TI - Changes in surface EMG and acoustic myogram parameters during static fatiguing contractions until exhaustion: influence of elbow joint angles. AB - The purpose of this study was to evaluate muscle fatigue using electromyogram (EMG) and acoustic myogram (AMG) signals of the shoulder and arm muscles during sustained holding tasks, with the elbow at different angles and at different levels of maximum voluntary contraction (MVC). The EMG and AMG of four muscles, including the upper trapezius (TP), anterior deltoid (DL), biceps brachii (BB), and brachioradialis (BR), were recorded during experiments using 10 healthy young males. The experiments were conducted under 9 pairs of conditions: 3 elbow angles (120 degrees, 90 degrees, and 60 degrees) and *3 levels of %MVC (20%, 40%, and 60%). Subjects were instructed to hold a weight equal to the designated %MVC at designated joint angles and asked to maintain that condition for as long as possible until exhaustion. Joint angles were also recorded by the electrogoniometers. The analysis of variance revealed that there was no significant effect of elbow angle on the mean MVC or on the endurance time. Elbow angle showed a significant effect on mean power frequency (MPF) of EMG in DL, BB, and BR, and a significant effect on root mean square (RMS) of EMG in four muscles. In BB and BR, MPF of EMG at 120 degrees was found to be significantly lower than 90 degrees and 60 degrees, respectively. There was a significant main effect of elbow angle on MPF of AMG for TP at 20% MVC; for DL at 20% and 40% MVC; for BB at 40% and 60% MVC; and for BR at the three levels of %MVC. The results showed that the range MPF of AMG for DL, BB, and BR was between 32 to 46 Hz, whereas that for TP was from 49 to 83 Hz. There was a significant effect of elbow angle on RMS of AMG in all four muscles in all experiments. At 20% MVC, a progressive increase in RMS of AMG was observed with time. In contrast, at 40% and 60% MVC, RMS showed very different behavior; specifically, it was found that RMS of AMG at 20% MVC significantly increased with increase of elbow angle. We conclude that RMS of AMG has a good and clear correlation with elbow angle at a low level of contraction. PMID- 11385937 TI - Relationship between sprint performance and muscle fascicle length in female sprinters. AB - The purpose of this study was to investigate the relationship between sprint performance and architectural characteristics of leg muscles in 26 female 100-m sprinters. Pennation angle and muscle thickness of the vastus lateralis (VL) and gastrocnemius medialis (GM) and lateralis (GL) muscles were measured by B-mode ultrasonography, and fascicle length was estimated. Sprinters had a significantly lower VL pennation angle, but GM and GL pennation angle was similar between sprinters and female control subjects (N = 22). There was no significant correlation between pennation angle and 100-m personal best performance. Sprinters had significantly greater absolute fascicle length in VL and GL than controls, which significantly correlated to 100-m best-record (r = -0.51 and r = 0.44, respectively). Relative fascicle length (VL and GL) were also significantly greater in sprinters than controls. However, there were no significant correlation between relative fascicle length and 100-m best-record (r = -0.36 and r = -0.29, respectively). No relationship was found between the sprint performance and fat-free mass (r = -0.26) or body mass index (r = -0.03). However, there was a significant correlation between percent (%) body fat and 100 m best-record (r = 0.62, p < 0.01). Adjusting the confounding effect of % fat, significant correlations were seen between relative fascicle length and 100-m best-record (VL; r = -0.39 and GL; r = -0.40). Absolute and relative fascicle length were similar in elite female sprinters compared with previous reported values for elite male sprinters (Kumagai et al., 2000). It was concluded that longer fascicle length is associated with greater sprinting performance in sprinters, but there is no gender differences in fascicle length for elite sprinters. PMID- 11385938 TI - Comparative stress in human societies. PMID- 11385939 TI - Electrodermal and cardiovascular manifestations of emotions in children. AB - The purpose of this paper is to analyze the current state of developmental researches in the area of psychophysiology of emotions in preschool and elementary school children. Electrodermal and cardiovascular activity measures are considered as the sources of indices of the autonomic nervous system activation during emotion-eliciting stimulation in children. We discuss the question of sensitivity of phasic and tonic autonomic measures for the identification of occurrence of emotion, mapping it along with valence and arousal dimensions in affective space, and to further differentiate emotions by their physiological manifestations. Considered are the conceptual and methodological issues related to psychophysiological measurements and developmental factors affecting the emotional reactivity in children. Special attention is devoted to the developmental aspects of psychophysiological studies on emotion such as the maturation of organs, integration of the autonomic and central nervous systems, age and gender-related changes in autonomic reactivity, and development of inhibitory control. Summarized are main findings relevant to psychophysiology of emotions in preschool and early school-age children and suggested are most perspective directions of their integration in the framework of modern theories of emotion. PMID- 11385940 TI - Intriguing topics in physiological anthropology: introduction to the theme. PMID- 11385941 TI - Virtual anthropology: the digital evolution in anthropological sciences. AB - The discovery and explanation of differences among organisms is a major concern for evolutionary and systematic biologists. In physical anthropology, the discrimination of taxa and the qualitative and quantitative description of ontogenetic or evolutionary change require, of course, the analysis of morphological features. Since the 1960s, a remarkable amount of fossil material was excavated, some of it still awaiting a detailed first analysis, some of it requiring re-examination by more developed methods. While the fossil record grew continuously, a revolution in anthropological research took place with advances in computer technology in the 1980s: a handful of innovative researchers working in specialized anthropology laboratories or medical departments developed the methodological inventory needed to extract critical information from subjects in vivo and from fossilized remains. A considerable part of this information is preserved in the physically heretofore inaccessible interior of anatomical structures. Virtual Anthropology (VA) is a means of making them visible and measurable. Thus, VA also allows access to 'hidden' landmarks; in addition, the large number of semilandmarks accessible on the form enhances the power of Geometric Morphometrics analysis. Furthermore, the density information in volume data allows manipulations such as segmentation, impossible with the real, physical object. Moreover, metric body measurements generally, and cranial measurements specifically, are also an important source of information for the analysis of the ontogenetic development of the skeletal system, and--last but not least--for clinical use (e.g., operation planning, operation simulation, prosthetics). Thus, there developed a fruitful interdisciplinary cooperation between statistics, medicine, and physical anthropology. PMID- 11385942 TI - Selenium status in an iodine deficient population of the West Ivory Coast. AB - Selenium is an essential trace element which is part of the active site of seleno dependent glutathione peroxidase and type 1 deiodinase. Therefore, it plays a key role in thyroid hormone metabolism. The present work was undertaken in order to evaluate selenium status in two Ivory Coast populations: the first with high (Glanle) and the second with low (Abidjan) prevalence of iodine deficiency. Selenium, glutathione peroxidase, glutathione reductase, glutathione and diglutathione were determined in blood and/or urine. In plasma and erythrocytes, selenium and glutathione peroxidase were dramatically low in Glanle. Compared to Abidjan, selenium, glutathione peroxidase, vitamin E and riboflavin status were decreased whereas diglutathione was increased in Glanle. The results clearly demonstrate a selenium deficiency and suggest an oxidant stress in Glanle. Causes and consequences of this selenium deficiency and oxidant stress remain to be determined. PMID- 11385943 TI - Endogamy and variation in blood pressure levels in Croatian island isolates. AB - Blood pressure variation was investigated among populations inhabiting islands and peninsula of Middle Dalmatia, Croatia. The number of previous anthropological studies pointed to isolation and different genetic population structure in this environmentally fairly homogeneous area. Variation in blood pressure (systolic and diastolic) among the populations of the islands of Brac, Hvar, Korcula, and the Peljesac peninsula was assessed at three levels involving village populations, regional (western and eastern) populations and the entire island populations. The blood pressure data were collected from 3834 adult individuals inhabiting 37 rural communities and were adjusted for age and body mass index. Variation in blood pressure levels existed among regions and villages. Due to the history of differential settlement, small village sizes and high levels of reproductive isolation, the observed blood pressure variation could be attributed to founder effect, genetic drift and inbreeding. The involvement of genetic factors was tested by relating blood pressure variation among villages to degree of isolation among them. Blood pressure means and proportions of hypertensives increased with endogamy levels in males. In females, this effect could not be observed. However, in both sexes the highest proportions of hypertensives (more than 40%) were found in villages that are most reproductively closed (endogamy greater than 80%). These populations are considered particularly promising for further genetic epidemiological research. PMID- 11385944 TI - The modernizing Kazakhstan: a review of biomedical data. AB - In order to focus the situation of Kazakhstan today in relation to the processes of modernization and transition to a market economy and to evidence their effects on the biology and health status of the population of Kazakhstan, we have reviewed recently available data for this region (1993-1999). Kazakhstan is still characterized by a pyramid shaped age distribution of its population and by a high incidence of not communicable diseases and lack of nutrient and micronutrients, especially among children. However, the population of Kazakhstan seems to be not immune to the diseases of the modernization. I.e., among women obesity is more frequent than underweight, especially in the urban areas. In rural populations the frequency of clinically relevant hypertension resulted low in the more isolated and traditionally living communities but it increased to 20% in the less isolated one. Although it is expected a strong increase of urbanized population in the next 25 years, currently, modernization is probably influencing life style and nutritional habits of almost only a minority of the inhabitants of Kazakhstan. PMID- 11385945 TI - E-procurement moves ahead. PMID- 11385946 TI - Stats & facts. Exorcising health care fraud. PMID- 11385947 TI - The financial implications of HMOs' partial county carve-out option. AB - This study examined the Health Care Financing Administration's policy allowing Medicare HMO risk contracts to carve out certain portions of counties from their service areas without adjusting the HMOs' capitation rate. In 1999, the policy resulted in 2.2 million Medicare enrollees losing access to HMOs and 2.3 million Medicare enrollees left with fewer HMO options. Although the majority of Medicare HMOs did not appear to be adopting the policy, there did seem to be a general tendency to exclude higher cost areas, which resulted in an estimated loss to the Medicare Trust Fund of $769 million in 1999. Of particular concern is the magnitude of profits and losses this policy generated for some individual HMOs. PMID- 11385948 TI - State and federal government mandates are changing the face of pharmacy management. PMID- 11385949 TI - Understanding disease prevalence in MCOs. PMID- 11385950 TI - The E-transformation of managed care: challenges precede rewards. PMID- 11385951 TI - Evaluating pharmacy benefit management information on the Internet: purpose, structure, technology, and content. AB - A wealth of health information is available on the Internet today. However, little is known about how to evaluate its quality and content. This exploratory analysis examined the Internet sites of 71 pharmacy benefit management firms that were operational in late 1999 by using components of several existing evaluation tools. Variables were used that incorporated interactive marketing strategy, technology features, and content. The principal findings suggest that health information on the Internet can and should be evaluated by users on dimensions such as purpose, structure, technology, and content. However, further work will be required to develop and test criteria. PMID- 11385953 TI - To trayline or not to trayline--that is the question. PMID- 11385952 TI - The future of pain management in managed care. AB - Pain management has made great strides in clinical practice and education, and in research documenting its effectiveness on pain relief, functional improvement, and cost. The profile of pain in managed care settings has recently been raised, notably because of the trend toward replacing inexpensive generic drugs with new and expensive drugs for pain treatment. Managed care organizations have greater motivation to focus on pain care because of recent regulatory initiatives, new accreditation standards, and legal precedents. In order for pain programs to improve the quality of pain care in MCOs, they must balance the complex needs of patients with pain with the financial constraints of the managed care setting. PMID- 11385954 TI - Making a difference with MNT and diabetes. Opportunity is knocking. PMID- 11385955 TI - Performance improvement. PMID- 11385956 TI - Applying the PDCA model. PMID- 11385957 TI - Tools for pretargeted radioimmunotherapy. PMID- 11385958 TI - Preclinical evaluation of a humanized NR-LU-10 antibody-streptavidin fusion protein for pretargeted cancer therapy. AB - A humanized single chain Fv antibody fragment specific to the EGP40 antigen was genetically engineered as a streptavidin fusion (scFvSA) for use in pretargeted radioimmunotherapy. The scFvSA construct was expressed as a soluble, tetrameric species in the Escherichia coli periplasm at 110-140 mg/liter. The fusion protein was purified from crude lysates by iminobiotin affinity chromatography with an overall yield of 50-60%. Characterization of the purified protein by SDS-PAGE, light scattering, and size exclusion chromatography demonstrated that the fusion protein was tetrameric with a molecular weight of approximately 172,000. Competitive immunoreactivity assays showed a two-fold greater binding to the antigen than the comparable whole antibody. The purified protein had a biotin disassociation rate identical to recombinant streptavidin and bound an average of three of four possible biotins per molecule. The radiolabeled fusion protein showed a faster blood clearance rate in normal mice than the corresponding whole antibody-streptavidin chemical conjugate. Tumor-specific targeting of a subsequently administered radionuclidechelate/biotin molecule was demonstrated in nude mice bearing SW1222 human colon carcinoma xenografts. A single dose of 800 microCi of 90Y-DOTA-biotin produced cures in mice with established subcutaneous human small cell lung or colon cancer xenografts. PMID- 11385959 TI - A pilot trial of Vitaxin, a humanized anti-vitronectin receptor (anti alpha v beta 3) antibody in patients with metastatic cancer. AB - The angiogenic response of a progressing malignancy is characterized by a shift in the balance of stimulatory and inhibiting factors of angiogenesis. Recognition of the regulated steps in tumor angiogenesis provides unique targets for developing anti-tumor therapy. Vitaxin is a humanized monoclonal antibody, which has specificity for the integrin alpha v beta 3 (vitronectin receptor). This antibody can impair the vascular response of endothelial cell growth factors in vitro and inhibit tumor cell mediated angiogenesis in pre-clinical animal models. Patients with metastatic cancer who failed standard therapy received intravenous doses of 10, 50 or 200 mg in cohorts of three patients. The unlabeled dose of Vitaxin was infused on days 0 and 21 of a treatment cycle. All patients received a pre-therapy imaging dose of 1 mg of Tc-99m Vitaxin with gamma camera imaging studies. There was no significant toxicity noted in these three dose levels. There were no objective anti-tumor responses. Three patients received two cycles of therapy and had stable disease at day 85 when taken off study. Radioimaging of tumor vasculature was unsuccessful although one patient with alpha v beta 3 positive melanoma had imaging of tumor sites. There was no immune response to Vitaxin in any patient. Patients receiving 10 mg doses of Vitaxin had poor plasma recovery of injected doses and brief circulation in plasma. Doses of 50 and 200 mg had plasma recovery that better approximated the predicted levels in plasma and circulation half-lives of approximately 7 days. This data suggests that an every three-week schedule of Vitaxin at doses of 200 mg (2.5-3.5 mg/kg) can maintain circulating levels of antibody with little or no toxicity. Future studies will be challenged to define anti-tumor activity in malignancy or appropriate surrogates of anti-tumor effect and explore escalating doses and alternate schedules of administration. PMID- 11385961 TI - Accumulation of radiolabeled anti-CEA antibody (mT84.66) in the case of multiple LS174T tumors in a nude mouse model. AB - A comparison was made between labeled antibody accumulations in nude mice having either single or multiple human xenografts. The LS174T tumors were implanted subcutaneously. All animals were given 2 micrograms of labeled murine anti carcinoembryonic antigen (CEA) monoclonal antibody 111In-mT84.66. Some animals were also given specific antibody pretreatment (SAP) of 200 micrograms of unlabeled mT84.66 to reduce liver accumulation of activity. In order to represent these multiple tumor examples, a simple initial-phase pharmacokinetic model was first fitted to each of the two groups (SAP and PBS treated) of single-tumor animals. Using the resultant six non-adjustable parameters as constants, the n = 1 uptake model was then used to represent tumor, liver and blood accumulations (%injected dose/organ) in the multiple-tumor animals. The model was found to be a good representation; in particular, it had far better agreement than single tumor predictions in the PBS mice. Differences between the single-tumor accumulations and those seen in multiple tumor examples were generally between two- and three fold. The model also demonstrated that the result of SAP was to essentially eliminate the effect of liver targeting of tumor-secreted CEA. We conclude that an initial-phase one-tumor model can describe the decrease of accumulation of activity in the case of multiple tumors in nude mice in both untreated (PBS) and pretreated conditions. Implications for clinical imaging and therapy with monoclonal agents are discussed. PMID- 11385960 TI - In vivo evaluation of bismuth-labeled monoclonal antibody comparing DTPA-derived bifunctional chelates. AB - Among the radionuclides considered for radioimmunotherapy, alpha-emitters such as the bismuth isotopes, 212Bi and 213Bi, are of particular interest. The macrocyclic ligand, DOTA, has been shown to form stable complexes with bismuth isotopes. The kinetics of the complexation of bismuth with the DOTA chelate, however, are slow and impractical for use with 212Bi and 213Bi that have half lives of 60.6 and 45.6 min. The study described herein compares six DTPA derived bifunctional chelates with the goal of identifying an alternative to the DOTA ligand for radiolabeling with bismuth. Radioimmunoconjugates comprised of MAb B72.3, each of the six DTPA chelates, and radiolabeled with 206Bi, which facilitated the evaluation due to its readily detectable gamma-emission. In vitro studies showed that each of the radioimmunoconjugates retained immunoreactivity that was comparable to its 125I-labeled counterpart. The 206Bi- and 125I-labeled immunoconjugates were then co-injected i.p. into normal athymic mice. Injection of Afree@ 206Bi demonstrated that the kidneys were the critical organ to evaluate for retention of bismuth in the chelate complex. Major differences were identified among the six preparations. The CHX-A and -B immunoconjugates were found to have 1) the lowest %ID/gm in the kidney; 2) a level of 206Bi in the kidney that was comparable to that of 125I-B72.3; and 3) no significant uptake of 206Bi evident in other organs such as bone, lung and spleen. The results described herein suggest that either of the cyclohexyl derivatives of DTPA may be suitable candidates for the labeling of immunoconjugates with alpha-emitting bismuth isotopes for radioimmunotherapeutic applications. PMID- 11385962 TI - Evaluation on a six-dose treatment of anti CD 20 monoclonal antibody in patients with refractory follicular lymphoma. AB - Treatment of refractory follicular lymphoma with monoclonal antibody CD 20 has been proven to be a good therapeutic option. However, most studies used four weekly doses and time to treatment failure (TTF) and overall survival (OS) could be considered very short: 11.0 and 13.6 months respectively. We started a pilot study to evaluate if six infusions at the same doses and schedule could improve the outcome in these patients. Seventeen patients with refractory follicular lymphoma heavily treated with chemotherapy (> 2 regimens), radiotherapy and biological modifiers were enrolled in a pilot study. They received 6 weekly doses, at 375 mg/m2, of monoclonal anti CD 20. In an intent to treat analysis, overall response was 76%, of which 47% (8 patients) were complete response and 5 patients were partial response. With a median follow-up of 28.6 months, 7 complete responders remain alive, free of disease, and 2 partial responses remain stable without additional treatment. Median to TTF has not been reached; yet, actuarial curves showed that at 3 years, 53% of patients are alive. The four patients who were failure died secondary to tumor progression. Overall survival (OS) at 3-year was 76%. Toxicity was mild, all patients completed the schedule on time and doses. The addition of two doses of anti CD 20 clearly improved OS and TTF in a group of patients with refractory follicular lymphoma heavily treated and with poor prognostic factors. However, the number is too short to draw definitive conclusions; more clinical trials are necessary to determine if 4 or 6 doses of anti CD 20 therapy are better in this setting of patients. PMID- 11385963 TI - The role of technetium-99m methoxyisobutyl isonitrile scintigraphy in suspected recurrent breast cancer. AB - The aim of this study was to determine the role of technetium-99m methoxyisobutyl isonitrile (99mTc-MIBI) scintigraphy in the evaluation of recurrence and metastases in breast cancer patients with mastectomy and/or radiotherapy. A prospective study was designed to assess the accuracy of 99mTc-MIBI scintigraphy in 36 patients (mean age 49 years) with suspected recurrent breast cancer. The scintigraphic studies were correlated with radiological findings and/or with histopathology. At 10-15 min after 740MBq 99mTc-MIBI injection, standard planar images were obtained in prone lateral and anterior supine views and then single photon emission computed tomography (SPECT) imaging was performed. A whole body imaging was also performed to demonstrate distant metastatic lesions. Totally 52 lesions were evaluated which 19 of them in 9 patients were malignant, while 33 lesions in 27 patients were benign. The sensitivity was 33%, 88% and the specificity was 96%, 93% for planar and SPECT imaging, respectively in loco regional lesions. Overall, the sensitivity and the specificity of MIBI imaging including whole body were 89%, 81%, in other conventional radiological imaging methods were 95%, 65%, respectively. 99mTc-MIBI scintigraphy using SPECT imaging may provide useful complementary information in patients with suspected recurrence breast cancer. PMID- 11385964 TI - Cyclophosphamide plus somatostatin, bromocriptin, retinoids, melatonin and ACTH in the treatment of low-grade non-Hodgkin's lymphomas at advanced stage: results of a phase II trial. AB - PURPOSE: Somatostatin, prolactin, retinoids, melatonin and ACTH have been shown to influence the lymphatic growth, and the action of the cyclophosphamide in lymphoproliferative disorders is well known. This provided the rationale to conduct, in patients with low-grade non-Hodgkin's lymphomas (NHL), a phase II trial of a combined association of cyclophosphamide, somatostatin, bromocriptin, retinoids, melatonin and ACTH. PATIENTS AND METHODS: Twenty patients with a diagnosis of low-grade NHL, stage III or IV, were included in this study. Patients received for one month the following treatment: cyclophosphamide, somatostatin, bromocriptin, retinoids, melatonin, and ACTH. The therapy was continued for two additional months in patients with stable or responding disease. After three months, the responding patients continued the therapy for three months and more. RESULTS: Twenty patients were assessable for toxicity and response; 70% (14 of 20 patients; 95% confidence interval [CI], 50% to 90%) had a partial response; 20% (4 of 20) had stable disease, and 10% (2 of 20) progressed on therapy. Going on with the treatment, none of the 14 patients with partial response had a disease progression (average follow-up time of 21 months, range, 7 to 25), and 50% of these patients had a complete response; among 4 patients with stable disease, 25% (1 of 4) had a partial response and 75% (3 of 4) progressed on therapy (mean time to progression [TTP] 14.3 months, range, 7 to 21). The toxicity was very mild, the most common side effects being drowsiness, diarrhea and hyperglycemia. CONCLUSIONS: The association of cyclophosphamide, somatostatin, bromocriptin, retinoids, melatonin, and ACTH is well tolerated and effective in treatment of low-grade NHL at advanced stage. PMID- 11385965 TI - Autoimmune liver disease. Current standards, future directions. AB - The diagnosis and management of autoimmune hepatitis continues to evolve as new diagnostic tests and new therapies are added to the armamentarium. Also encouraging are the advances in the understanding of the human immune system and its involvement in the origin and course of auto immune diseases in general and in the variants of autoimmune liver disease. Promising changes are expected in the next few years as new medications become available to the practicing hepatologist. New immune tests may allow therapies to be customized to patients, and antiviral therapies may also eventually be used in the management of this autoimmune liver diseases. PMID- 11385966 TI - Hepatic fibrosis. Pathogenesis and principles of therapy. AB - There has been great progress made in our understanding of the cellular mechanisms of hepatic fibrosis. The recognition that the hepatic stellate cell, (formerly know as lipocyte, Ito, or fat-storing cell), played a central role in the fibrotic response was key to our understanding. Stellate cells undergo a process known as activation, in response to any insult. Activation is a broad phenotypic response, characterized by distinct functional changes in proliferation, fibrogenesis, contractility, cytokine secretion, and matrix degradation. Insights gained into the molecular regulations of stellate cell activation may lead to new antifibrotic therapies, which may reduce morbidity and mortality in patients with chronic liver injury. PMID- 11385967 TI - Current and future therapies of hepatitis C. AB - With recent advances in the treatment of chronic hepatitis C, patients with elevated aminotransferase levels, detectable HCV RNA in the serum, and chronic inflammation are candidates for therapy. The best initial therapy is interferon plus ribavirin, achieving a sustained response rate in 40% of patients. The duration of therapy should be based on HCV genotype (48 weeks for genotype 1; 24 weeks for other genotypes). Serum HCV RNA should be measured at week 24 to assess response and guide further therapy in patients with genotype 1 infection. Patients unsuitable for combination therapy can be treated with interferon monotherapy. Side effects, dose modification and discontinuation are generally more frequent with interferon plus ribavirin, but can be managed with close follow-up and careful monitoring. With rapid developments in treatment, new therapies will require careful prospective evaluation according to HCV genotype and viral-load characteristics. Recommendations for therapy will probably change every few years, and novel approaches may provide effective therapy for most patients with hepatitis C. PMID- 11385968 TI - Hepatitis B infection in China. AB - Chronic HBV infection is a serious health threat in the Asian-Pacific region. The introduction of lamivudine has greatly improved the hope of these patients and is undoubtly a milestone in the management of chronic HBV infection. The combination of lamivudine with another nucleotide or nucleoside analogue or immunomodulatory agent to improve its therapeutic efficacy further must be investigated. Also, the use of lamivudine to prevent HBV reactivation on withdrawal of immunosuppressive therapy should be explored. PMID- 11385969 TI - Molecular therapeutics of liver disease. AB - Fundamental advances in biomedical research will revolutionize the prevention and treatment of liver disease during the early twenty-first century. Recent progress in gene-, cell-, and recombinant protein-based therapeutics will contribute to this revolution, although formidable obstacles currently prevent the clinical application of these novel therapies. Eventually, these obstacles will be overcome, and molecular therapeutics of liver disease will become a clinical reality. As a result, the new millennium will be a very interesting time to practice hepatology. PMID- 11385970 TI - Artificial liver support devices for fulminant liver failure. AB - Artificial liver-support devices attempt to bridge patients with fulminant hepatic failure until either a suitable liver allograft is obtained for transplantation or the patient's own liver regenerates sufficiently to resume normal function. It is thought that toxins contribute to the clinical picture of fulminant hepatic failure. The earliest reports of successful toxin removal were blood- and plasma-exchange transfusions. Given these successful case reports, mechanical liver-support devices were designed to filter toxins. These mechanical devices used hemodialysis, charcoal hemoperfusion, hemoperfusion through cation exchange resins, hemodiabsorption, and combinations of all of these techniques as in the MARS liver-support device. Despite promising case reports and small series, no controlled studies of mechanical devices have ever showed a long-term survival benefit. Thus, the removal of presumed toxins seems to be insufficient to support patients with fulminant hepatic failure, and the biologic function of the liver must also be replaced. Attempts at replacing the biologic function have included extracorporeal liver perfusion, cross-circulation, and hepatocyte transplantation. Current technologies have combined mechanical and biologic support systems in hybrid liver-support devices. The mechanical component of these hybrid devices serves both to remove toxins and to create a barrier between the patient's serum and the biologic component of the liver-support device. The biologic component of these hybrid liver support devices may consist of liver slices, granulated liver, or hepatocytes from low-grade tumor cells or porcine hepatocytes. These biologic components are housed within bioreactors. Currently the most clinically studied bioreactors are those that use capillary hollow-fiber systems. Both the bioartificial liver by Demetrious and the extracorporeal liver assist device by Sussman and Kelly are in clinical trials. Although the trials seemed to have yielded good survival data when the devices are used as a bridge to transplantation, the type and degree of liver support provided by these devices remains uncertain. Thus, despite decades of great progress in the field of artificial liver support, no one technique alone yet provides sufficient liver support. A hybrid system seems to be the best option at present. Still to be determined is the best tissue to use, how much liver tissue should be used, and the optimal design of the bioreactor. PMID- 11385971 TI - Liver transplantation. Living donor, hepatocyte, and xenotransplantation. AB - Liver transplantation is now accepted as effective therapy in the treatment of acute and chronic hepatic failure. Improvements in surgical techniques and immune suppression have led to 5-year survival rates that exceed 70% in most centers. The success of transplantation has led to a dramatic increase in the number of candidates to over 14,000 places on the national waiting list. While the number of patients in need of transplantation increases, there has been little growth in the supply of available cadaveric organs, resulting in an organ shortage crisis. With waiting times often exceeding 1 to 2 years, the waiting list mortality now exceeds 10% in most regions. Several novel approaches have been developed to address the growing disparity between the limited supply and excessive demand for suitable organs. PMID- 11385972 TI - Herbal and complementary and alternative medicine therapies for liver disease. A focus on Chinese traditional medicine in hepatitis C virus. AB - This article presents herbal and alternative therapies used in the treatment of liver disease, both as adjunctive or complementary treatment to Western pharmaceutical strategies, and as an alternative treatment in liver disease, particularly for Hepatitis C Virus. There is a special emphasis on traditional Chinese herbal medicine. PMID- 11385973 TI - Liver cancer. AB - The prognosis of patients with HCC remains dismal. Even in the subgroups of patients who have the most favorable characteristics and are eligible for surgical resection, the 5-year survival rate is less than 25%. For patients with more advanced disease, the median survival time is less than 1 year. The good news in HCC research is that the disease can be prevented. In Taiwan, the rate of HCC in children aged 6 to 9 years decreased from 5.2 per million population before the neonatal vaccination program began in 1984 to 1.3 per million population in the first vaccinated cohort. Treatment of viral hepatitis with IFN may decrease the rates of long-term development of HCC. Other agents that may prevent second primary tumors following resection of HCC, such as polyprenoic acid and acylic retinoid, are also being investigated. PMID- 11385974 TI - Portal hypertension. AB - Many advances in the management of portal hypertension and variceal hemorrhage have occurred during the last 10 years. Effective therapy for primary prevention of variceal hemorrhage is now available in the form of nonselective beta blockers. Active bleeding should be managed with terlipressin, somatostatin or its analogues, and endoscopic therapy; TIPS and surgery are reserved as salvage therapy for patients who fail endoscopic treatment. Survivors of a variceal hemorrhage should be evaluated for liver transplantation. Specific treatment may be provided with EVL while these patients await transplantation. Patients who fail endoscopic treatment may be treated by TIPS or surgery. PMID- 11385975 TI - Management of ascites. AB - The evaluation of ascites includes a directed history, focused physical examination, and diagnostic paracentesis with ascitic fluid analysis. Dietary sodium restriction and oral diuretics are the mainstay of therapy for the majority of patients with cirrhotic ascites. Transjugular intrahepatic portocaval shunt has emerged as the treatment of choice for selected patients with refractory ascites, although serial large-volume paracenteses should be attempted first. Early diagnosis, broad-spectrum antibiotics, and albumin infusion contribute to the successful management of spontaneous bacterial peritonitis (SBP). Referral for liver transplant evaluation should be considered at the first sign of decompensation and should not be delayed until development of ominous clinical features, such as refractory ascites and SBP. PMID- 11385976 TI - [On some basic issues and factorial structure of ecological pulmonology]. AB - Consideration is given to the place occupied by diseases of respiratory organs in the general structure of morbidity. In many ways, growth in their number has been determined by amplification of anthropogenic factors. This is manifested in emergence of new eco-dependent diseases and syndromes including more difficult course of pulmonary pathology. One of the prioritized disciplines of the modern pulmonology is ecological pulmonology which lies in the domain of medical ecology and is central to the concept of eco-dependent human diseases. A factorial structure of ecological pulmonology built on the etiological principle is proposed to facilitate inquiry into the origin and course of various forms of eco dependent pathology of the lung. PMID- 11385977 TI - [Simulation of the dynamics of subjective status in an extended flight aboard a single-seat airplane]. AB - Based on experimental data regarding changes in the subjective status of 29 operators during 12-hr simulation of a flight in a single-seater airplane, individual parameters were determined for a mathematical model of the well-being and anxiety dynamics. The model was devised to reproduce two processes, i.e. the pre-launch reaction and effects of a set of "flight" factors that displayed either exponential or S-form dependence. Analysis showed that the pre-launch reaction was distinctly linked with well-being in 15 individuals and with anxiety in 11 individuals. Well-being in one individual and anxiety in seven individuals did not reveal any regular trend. Standard deviations in the evaluation of well being and anxiety were within the range of 0.08-0.47 and 0.4-3.3 scores, respectively, that is did not exceed 6-8% of the maximal possible range of variation. On the whole, the mathematical model gave an accurate description of the dynamics of subjective status. PMID- 11385978 TI - [Age-related characteristics of osteopenia in rats caused by deficient support loads on hind limbs]. AB - Progress of osteopenia in 40-, 100-, and 220-d old rats who had been suspended by tails to release support loading on their hind limbs for 30 days was compared. Deficient support loading was shown to inhibit the longitudinal growth of tibia and produce osteopenia of the proximal metaphysis spongiosis in all groups of rats. Humerus of the rats whose front limbs were still partially loaded, suffered same changes as tibia though much less pronounced. Inhibition of bone growth in length and osteopenia in bone spongiosis reached maximum dimensions in 220-d old rats as a result of summation of kindred changes due to the load deprivation, and aging. Established was a correlation between suppression of the functional activity of STH-cells in adenohypophysis and progress of bone atrophy during suspension. Functional activity of thyroid calcitonin-producing cells was also reduced with the maximum decline in the 40-d old rats. PMID- 11385979 TI - [Peculiarities of development of muscular atrophy in rats of varying age due to suspension]. AB - Compared were changes in m. soleus and m. gastrocnemius in rats at the age of 40, 100 and 220 days in a 30-d experiment with tail suspendion. As it was stated, deficient support loading of the hind limbs had a negative effect on the gain in m. soleus mass and, to a less degree, in m. gastrocnemius mass, particularly in younger rats. In tail-suspended young rats m. soleus mass was by 60% and in elder rats by 48% less as compared to their controls. No matter age of the group, type 1 fibers and IIA and IIC in m. soleus were equally affected. In comparison with the controls, cross-section areas of fibers in 40- and 100-d old rats decreased by 62-68% and in 220-d old rats, by 43-50%. Slow type 1 muscular fibers underwent transformation in fast type IIA and IIC fibers in m. soleus. The more aged rats, the less population of slow muscular fibers transformed. The m. gastrocnemius mass in all suspended groups was by 20% lower than in the controls. In 40- and 100-d old rats suspension caused atrophy of each of 5 types of fibers in m. gastrocnemius, whereas in 220-d old rats those were only types IA and IIC of fibers that had been atrophied. There were early signs of transformation of slow type IB fibers in the fast ones. The conclusion was made that muscles of young animals are more sensitive to deficit in functional loading and atrophy more severely that in old rats. Transformation of slow fibers was observed mainly in m. soleus of young animals and was essentially absent in old. PMID- 11385980 TI - [Effect of antiorthostatic hypokinesia and its combination with +Gz runs on the otolith regulation of cardiac rhythm in primates]. AB - Amplitude of cardiac rhythm response to the otolith stimulus (AR-R), i.e. rapid reduction of the EKG R-R interval was determined in 24 Macacas mulatta before and after 28-d head-down (-6 degrees) immobilization (HDI). In two experimental series the animals were divided into 2 groups (control and CF) with 6 primates in each. In the HDI series-1, the CF animals were rotated on a centrifuge with a 3.62-m arm at 1.2 up to 1.6 +Gz for 30-40 minutes 4-5 times a week. In HDI series 2, the CF animals were rotated 30 min at 1.2 +Gz 2-3 times a week. In a day after HDI, AR-R was significantly reduced in both CF groups, which, when compared with EKG R-R intervals, suggests attenuation of equally parasympathetic and sympathetic influences on the heart. In 7 days since HDI, AR-R was close to baseline values in control primates on both series and CF primates in series-2, whereas in the CF group in series-2 AR-R exceeded baseline values. Significant differences in mean AR-R values in the control and CF groups in series-1 following 7 days after HDI point to the prophylactic effect of small but regular g-loads manifested in more rapid recovery of initial level of the cardiac rhythm regulation. PMID- 11385981 TI - [Alterations in actin cytoskeleton and rate of reparation of human endothelium (the wound-healing model) under the condition of clinostatting]. AB - Effects of long-term simulation of hypogravity on actin cytoskeleton and cell migration were investigated in cultured human endothelium cells (EC). In control, F-actin resided predominantly on the periphery of cell forming an array of parallel bundles with "dense bodies" along the edge. A small number of actin cable fibers was found in the center. Already after 1-2 hrs of clinostatting at 5 RPM the cell cytoskeleton showed actin filament thinning and displacement toward the cell edges. In subsequent 6-18 hrs, almost all actin fibers had left the center part of EC and had ranged themselves in a continuous F-actin line in the intercellular contact area. In most cases, these changes resulted in the so called "ruff-edge". Since both the disappearance of cable fibers and formation of the "ruff-edge" add to the cell migration activity, this parameter was studied with the would-healing model. According to our data, 24-48 hrs of exposure to hypogravity stimulates cell migration and expedites 2-3 times reparation of mechanically damaged monolayer. The results suggest that effects of hypogravity on cultured human EC are likely to be consequent to alterations in the activity of protein kinase C and/or adenylate cyclase involving many members of the cellular metabolism. PMID- 11385982 TI - [Reactivity of the arterial system to orthostasis]. AB - Reported are data of experiments with unconscious tilted rats (30 degrees, 45 degrees and 60 degrees) in which a dependence of orthostatic hypotensive reactions of systolic (APs) and diastolic (APd) pressure on spontaneous level of baseline mean arterial pressure (APb.mean) was established. Rats tilted at 30 degrees and 45 degrees, APs and APd were in inverse linear dependence with APb.mean above 95 mm Hg and in direct dependence with APb.mean below 95 mm Hg. Pressor effects of phenylephrine correlated directly and to the same extent with APb.mean only below 95 mm Hg in the 30 degrees and 45 degrees tilt and both above and below 95 mm Hg in the 60 degrees tilt. Mean values of the pressor effect before and after experiment were different in animals tilted at 60 degrees. No dependence of cardiac output on APb.mean was determined. Therefore, the trend and depth of changes in the arterial system reactivity to tilt are dependent on the degree of inclination and the APb.mean range. Reactivity to the adrenergic agent alters only when inclination is greater than 45 degrees. PMID- 11385983 TI - [Fat loading test: biochemical criteria and informative value of the physical endurance assessment]. AB - Biochemical criteria of the fat loading test (FLT) were established and the search for interrelations between FLT biochemical variables and physiological parameters of dynamic physical testing (bicycle ergometry) was conducted in the experiment with participation of male volunteers. Relative gain in glucose, triglycerides, total cholesterol concentrations in blood, atherogenicity factor and insulin decrease after 3-hr FLT and reduction in blood concentrations of total cholesterol, insulin, hydrocortisone and testosterone after 9-hr FLT were ranked as reliable criteria. Rise in blood glucose and decrease in blood insulin after 3-hr FLT, and decrease in serum testosterone after 9-hr FLT were found to have prognostic value. Correlation between the FLT biochemical variables and physiological parameters as well as the package of regression analysis equations allow prediction for physical endurance. Yet, analysis of the correlation suggested that desensitization to insulin of the cell membrane cells should be a pathogenic mechanism of physical endurance decrement. PMID- 11385984 TI - [Psychosocial aspects of perception of technology-related risk]. AB - In addition to the somatic effects, physical and chemical factors may also have sociopsychological impacts and distort perception of technogenic risks. Risk management is an important component in the analysis of human-environment interaction. Individuum per se with her/his system of values plays a key role in risk perception and transformation of this perception. Reviewed are the sociopsychological constituents of risk perception that often give birth to phobias, as well as national-ethnic peculiarities and the role of mass media in provoking phobias. Emphasized is the necessity to continue studies of phobias evoked by technogenic risks in the next areas: somatic and mental health and risk perception, genetic mechanisms of resistance to physical and toxic factors, ethnic and national factors in risk perception, sociopsychological aspects of interaction of novel technologies, society, public health and ecological movements; ideology, religion, mysticism, mass-media: formation of distorted perception of risk; risk management. PMID- 11385985 TI - [The assessment of the risk of altitude decompression sickness during simulation of repeated (with 12 hour interval) exits into open space]. AB - Purpose of the investigation was to assess contribution of repeated (with a 12-hr interval) decompression to the risk of altitude decompression sickness (ADS) by simulation of 6-hr extravehicular activities (EVA) of space crewmembers in altitude chamber. The protocol included "ascents" of 6 essentially healthy male subjects at the age of 24 to 51 to the altitude of 7,600 m (37 kPa) following 30 min prebreathing (elimination of nitrogen from the body by breathing pure oxygen through a mask at the ambient pressure of 73 kPa = 2,600 m). Each subject participated in 2 experimental exposures: first initial and then repeated decompression. None of 24 "ascents" produced clinical signs of ADS. Comparison of the data concerning frequency and time points of detection by ultrasonic Doppler equipment of gas bubbles (GB) in the venous bed during decompression with initial, maximal and mean values of US signal intensity failed to state a significant difference between them. Data of the investigation were confronted with anticipated length of GB dispersion in body tissues. PMID- 11385986 TI - [The effect of complex barometric exposure on hemodynamics during simulation of the physiological impacts microgravity in animals]. AB - In experiments with anesthetized cats effects of breathing under negative pressure (BNP, -5 cm of water column) combined with lower body negative pressure (LBNP, -20 cm of water column) on the cardiorespiratory reactions to postural simulation of the hemodynamic shifts in microgravity were studied. Evidence was obtained that this complex barometric exposure of tilted animals (-30 degrees) imitated the central and peripheral hemodynamics characteristic of tilt at the calculated inclination of +6 degrees up to +12 degrees. Extrapolation of the experimental data on the orthodox physiological model of microgravity (HDT, -6 degrees) allows an assumption that the protocol of complex barometric exposure tested in this experiment transforms the hemodynamic parameters under study to the levels close to those in the vertical body. Results of the investigation infer an additive character of the complex barometric exposure (LBNP + BNP) and its utility as a model of orthostatic g-loads. PMID- 11385987 TI - [Development of a cultivation technology and selection of leaf vegetable cultures for space greenhouse]. AB - We plan to perform space experiments on development of a technology for cultivation of leaf vegetables that might be a component of future life support systems for space crews. For this purpose, we are going to fly research greenhouses with the crop area from 0.03 up to 0.1 m2 inside the universal docking module of the ISS Russian segment. To prepare for future space experiments, ground investigations were made in order to compare crop capacity of various artificial soils used to grow leaf vegetables in greenhouse Svet. Useful life of root module Svet can be extended with a new technology based on resupply of fibrous substrate with nutrients. The most effective volume of soil per a plant was determined which sustains high productivity of leaf vegetables in Svet. To select leaf vegetables for in-space cultivation, we conducted investigations of productivity, morphometric and biochemical characteristics, and palatability of 18 cultures including alternative greens highly popular in Japan and China which have been earlier tested neither in laboratory nor in space within the closed eco-system projects. We would prioritize mizuna, pak choi, tatsoi, rapina or broccoli raab, and red giant mustard as objects for in-space investigations. PMID- 11385988 TI - [The glutathione-dependent antioxidant system in rats during acute intoxication by hydrazine]. AB - Intensity of lipid peroxidation in the liver, kidney and brain tissues, and functioning of the glutathione system were studied in purebred rats after acute intoxication by 1.1-dimethyl hydrazine at the dose of 104.5 mg/kg of body mass (LD50) and phenyl hydrazine at the dose of 170 mg/kg of body mass (LD50). According to results of the investigation, the character and direction of changes in LPO and the antioxidant system due to intoxication by hydrazines are materially dependent on their structure and affected organs and tissues. For instance, intoxication by phenyl hydrazine markedly activates free-radical activation in all analyzed tissue with accumulation of malone dialdehyde against depletion of reduced glutathione in tissue and deactivation of enzymes involved in the antioxidant defense. Intoxication by dimethyl hydrazine was associated with moderate activation of free-radical oxidation in liver with a downward LPO trend in kidney and brain at the cost of exaggerated activity of enzymes in the glutathione system and maintenance of sufficient reserve of reduced glutathione. Hence, aryl hydrazine derivatives possess more distinct pro-oxidant properties. PMID- 11385989 TI - [Antioxidant effects of vetoron in flying personnel with arterial hypertension]. AB - The purpose of the investigation was to assess the possibility to control free radical oxidation in patients with cardiovascular pathologies with the help of vetoron. This was the first attempt to study effects of a beta-carotene (vetoron) on lipid peroxidation and the activity of antioxidant defense enzymes in flying personnel with cardiovascular pathologies (hypertension and neurocirculatory dystonia). Therapeutic effect of the medicament was evaluated by its action on peroxide metabolism (malonic dialdehyde, acyl hydroperoxides and trien conjugates, Schiff bases) and stimulation of the antioxidant defense enzymes (catalase and glutathione reductase). Results of the investigation pointed to the inhibition of lipid peroxidation against activation of the antioxidant defense enzymes in patients with hypertension who had been given the course of vetoron; however, in patients with neurocirculatory dystonia. PMID- 11385990 TI - [Methods of urine analysis for creatinine after long period of storage]. AB - Our investigations showed that n-cresol (4-methylphenol) stabilized urine and preserves urinal creatinine as long as for 50-d storage at room temperature. This antimicrobial biochemical stabilizer is the core of a novel method of delayed creatinine quantification in urine. It can be useful in metabolic investigations of cosmonauts and nuclear submarine crews, members of alpine, desert, deep-water, Arctic and Antarctic expeditions and in many other cases when samples cannot be analyzed immediately. PMID- 11385991 TI - [Evaluation of gene protective effects of some natural anti-mutation substance]. AB - Antimutation activity of some bee products was evaluated with a battery of test systems. Tablets of apilac prepared out of queen bee milk did not demonstrate gene-protective effects in Ames' test. Other bee products, i.e. propolis, beebread, honey preparations 1 and 2 and queen bee milk were able to mitigate to a degree ill effects of some chemical and physical mutagenes. Propolis was found to be toxic to the yeast cell. Antimutation properties of natural queen bee milk revealed in the experiment with test-objects were lost after the product had been processed into the tabloid form or stored at room temperature. PMID- 11385992 TI - [Inborn deficiency of complement C4A and C4B isotypes in persons infected with Chlamydia]. AB - The difference in the functional activity of the isotypes A and B of component C4 of human complement was used to determine their ratio to detect the inherited deficiency of the isotypes. The frequency of deficiency in healthy persons blood donors was equal for C4A and C4B (0.14 for each isotype), i.e. 14% of total number (22) donors, or 28% totally. These results agree with the literary data, on which the frequency of deficiency of C4A is 0.14, and of C4B is 0.11-0.16. The inherent deficiencies of C4A and C4B for persons infected by Chlamydia were studied. For this purpose the patients (35 persons) with in blood antibodies (IgG or IgM) to Chlamydia (C. trachomatis, C. psittaci and C. pneumoniae) were investigated. The frequencies of deficiency of C4A and C4B were 0.29 and 0.46 respectively. Thus, the number of the undeficiency patients was only 25%, while among healthy persons 70-75% of individuals not having deficiencies of isotypes C4 were observed. The deficiencies of isotypes of C4 at this pathology is detected for the first time. The obtained data suggest the existence of the predisposition to the development of diseases stipulated by Chlamydia in persons with inherent deficiency of C4 component of complement. PMID- 11385993 TI - [Isolation of glutamylendopeptidase precursor from B. licheniformis and its processing in vitro]. AB - A secretory system based on L-form cells of Proteus mirabilis was developed for production of native Bacillus licheniformis glutamylendopeptidase precursor never formerly available. The produced precursor was stable per se under physiological conditions and in presence of trypsin and glutamylendopeptidase from B. intermedius. Complete conversion of the precursor to the mature glutamylendopeptidase was performed by bacillar metalloproteases and subtilisin. The artificially processed glutamylendopeptidase was purified by affinity chromatography on bacitracin-sepharose. Native tertiary structure in the purified glutamylendopeptidase was confirmed by demonstrating its activity towards a specific chromogenous peptide substrate. PMID- 11385994 TI - [Deletion analysis of the structural-functional organization of metalloendopeptidase precursor in B. amyloliquefaciens]. AB - Functional destination of propeptides and precursors in bacillar secretory proteases remains uncertain. Formerly deletion assay demonstrated folding and secretion of subtilisin E, chymotrypsin-like protease SGPB from S. griseus and B. cereus metalloprotease to depend on full-length propeptide in the precursors. Actually an artificial B. amyloliquefaciens metalloprotease gene with deletion of 51 amino acid residues from N-terminus was constructed with regard to carry out functional mapping of secretory metalloprotease propeptides. B. subtilis wprA gene 5'-terminal region spanning promoter and secretory leader was coupled to provide transcription to the truncated gene and secretion to its product. B. subtilis clones bearing a plasmid with the modified gene synthesised an active mature metalloprotease. PMID- 11385995 TI - [Inhibitors of proteolytic enzymes in the treatment of diabetes mellitus]. AB - A new approach to overcome the degradation of insulin by proteolytic enzymes and its targeting to the blood through the digestive apparatus was developed. The approach is based on the immobilization of insulin into the polymeric hydrogel which is modified by ovomucoid--glycoprotein, inhibitor of proteolytic enzymes. Oral administration of this system to rabbits and rats, (in contrast to the hydrogels modified by proteolytic enzymes inhibitors without polysaccharide part), statistically significantly lowered blood glucose level. PMID- 11385996 TI - [Kallikrein-kinin system: novel facts and concepts (literature review)]. AB - The kallikrein-kinin system (KKS) is the key proteolytic system participating in control of a wide spectrum of physiological functions and the development of many pathological conditions. This explains great interest in structures, functions and molecular biology of separate components of the system, molecular mechanisms of their interaction and relationship with other regulatory systems. The information in this field for the last two decades clarifies the role of KKS in morphogenesis of cells, regulation of smooth muscular contractility of some organs, decrease of blood pressure, increase of vascular permeability, the development of inflammation, transformation of cells and the other functions of both physiological and pathological processes. Essential progress in understanding of functions KKS was made by the discovery and study of bradykinin receptors, cloning of kininogen and kallikrein encoding genes, revealing of domain structure of kininogen, prekallikrein and some kininase and decoding of mechanisms of contact phase of proteolytic system activation in blood plasma. PMID- 11385997 TI - [Energy-dependent selective intracellular proteolysis. Structure, active sites and specificity of ATP-dependent proteinases]. AB - The enzymatic systems of selective proteolysis serving for the maintenance of cell homeostasis and functioning in prokaryotic and eukaryotic cells are characterized. The data on structure, active sites and specificity towards protein targets are given. PMID- 11385998 TI - [Angiotensin-converting enzyme and its physiological role]. AB - Angiotensin-converting enzyme (ACE, peptidyl dipeptidase A) is well known as a key enzyme involved in regulation of blood pressure. Regulation of blood pressure is the main but not the only ACE function. This enzyme is also involved in the regulation of a range of other physiological processes including control of cells proliferation. Particular role of the enzyme in a given process is determined by its localization and its action on regulatory peptides. Structural peculiarities of angiotensin-converting enzyme and its gene polymorphism related to its physiological function are discussed. ACE molecule consists of two large homologous domains (N- and C-domains), differing in their catalytical properties. These differences are suggested to be physiologically important. Endogenous substrates specific for each of these domains and natural active single-domain ACE forms were revealed. An association between the ACE gene polymorphism and plasma ACE levels, as well as with an increased risk of various cardiovascular and renal diseases was observed. PMID- 11385999 TI - [The effect of leukocyte elastase on high molecular weight kininogen from human plasma in the presence of alpha-1 protease inhibitor. Analysis of proteolytic degradation]. AB - Degranulation of polymorphonuclear leukocytes (neutrophils) and releasing of leukocyte elastase during inflammation occur not only in injured tissue but in plasma in the presence of considerable excess of alpha-1 proteinase inhibitor (alpha-1PI). However, in spite of the absence of free elastase in patients' plasma, even in such severe inflammation as peritonitis and septicaemia, degradation of the connective tissue structures and plasma proteins may be determined. However the reasons of such destructive action are not yet determined. In this paper the action of leukocyte elastase on human plasma high molecular weight kininogen (HMWK) was studied in the absence or in the presence of different concentrations of alpha-1PI. The results showed that degradation of the intact molecules of HMWK occurred under the action of elastase during 1-2 hours of combined incubation even if the concentration of alpha-1PI in the mixture in 3-5 fold exceeds the molar elastase concentration. The rate of elastase inhibition by alpha-1PI in the presence of HMWK did not depend on an order of enzyme and inhibitor addition to the incubation medium. HMWK degradation by elastase in the presence of alpha-1PI was accompanied by impairments in its adhesion function although high tolerance of HMWK inhibitory activity with respect to SH-proteinases preserved. Thus, total inhibition of leukocyte elastase by alpha-1PI, in the presence of high molecular weight kininogen develops during relatively long time interval. The pronounced destruction of intact HMWK molecules takes place during this period of gradual elastase inhibition. This fact seems to be very important in pathogenesis of thrombo-haemorrhage syndrome as a complication of severe inflammation. PMID- 11386000 TI - [Type I and IV collagenases and their endogenous regulators in immortalized and transformed fibroblasts]. AB - To elucidate the role of matrix metalloproteinases (MMP) in carcinogenesis, the expression of collagenases of types I (MMP-I) and IV (MMP-2 and MMP-9) as well as the behaviour of urokinase-like plasminogen activator (uPA) and of tissue MMP inhibitors (TIMP) in immortalized (IF) and transformed (TF) fibroblasts were investigated. The study was carried out using embryo rat fibroblasts, sequentially immortalized with the LT gene of human papilloma virus and transformed with the E7 gene of human papilloma virus (HPV-16). As control was used the primary fibroblast (PF) culture of Fisher rats. In IF, the collagenase activity was at the same level as it was in PF. The activity of uPA in IF was increased by 2-2.5-fold; the titrated amount of free endogenous inhibitors in IF and PF was at essentially the same level while being markedly higher than in TF. At the stage of fibroblast transformation with the E7 gene of HPV-16, there was seen an increase of Type IV collagenases and a decrease of Type I collagenase, both these indices being most pronounced in the cells with most developed tumorigenic properties. In TF there occurred a decrease of free endogenous MMP inhibitors relative to the enzyme activity and, at the same time, a decrease in uAF activity, indicating the changes occurring in the enzyme/inhibitor/activator ratio and hence the enhancement of the destructive potential of the cells (in this case, at the cost of Type IV collagenase activity). PMID- 11386001 TI - [Turbidimetric analysis of fibrin polymerization in the plasma]. AB - The turbidimetrical assay of thrombin-induced plasma coagulation provides a possibility to estimate both stages of fibrinogen-fibrin conversion. The initial one, which proceeds without any change of turbidity, reflects the process of protofibril formation, and the second stage of lateral aggregation, is characterized by the rise of turbidity. The influence of heparin, alga (Laminaria digitata) aqueons extracts, and collagenase on the indices of the turbidity-time curve has been studied. It was established that the alga extracts possessed the powerful heparin-like anticoagulant activity. The both agents influenced the first stage of the turbidity-time curve, suppressing protofibril formation, which reflects the thrombin inhibition. Nevertheless, they differed in their mode of dose-dependence. While the time of protofibril formation was direct proportional to the alga extract concentration, it was rising more intensively with heparin dose elevation. Plasma pre incubation with alga extract or heparin did not influence their action. Treatment with plasma collagenase changed only the second stage of the coagulation curve. It inhibited the process of protofibril lateral aggregation in the direct proportional manner. It must be due to fibrin digestion by the enzyme. We propose that fibrin cleavage by collagenase occurred out of the thrombin action sites, because the velocity of protofibril accumulation stayed unchanged. Our data illustrate the usefulness of the turbidimetrical analysis in the studies of the agents' action mechanisms on blood coagulation, in conditions close to physiological ones. PMID- 11386002 TI - [Modification of proteolytic complement cascade after treatment with exogenous heparin]. AB - The influence of heparin therapeutic concentrations (0.25-10 U/ml) on the kinetic indices of human serum complement activity has been studied. An acceleration of complement-dependent lysis of rabbit erythrocytes via alternative complement pathway was found at low (0.25-0.5 U/ml) heparin concentrations and complement inhibition at higher (5-10 U/ml) concentration. The alternative pathway inhibition was revealed mainly by lag-period elongation of the hemolysis. Heparin in the concentrations more then 1 U/ml caused dose-dependent lowering of sheep erythrocytes hemolysis rate accompanied with reciprocal elevation of complement hemolytical capacity, which shows more "economical" consumption of complement components at heparin presence. The identical effect was observed in sera, deficient in factor D of alternative pathway of complement activation. This can be an evidence of the prevailing heparin inhibition of the classical pathway. PMID- 11386003 TI - [Peptidyl hydrolase activity of blood serum in women with neoplastic diseases of the endometrium]. AB - Blood serum proteinase activity was studied in healthy women and in women with endometrial tumors. Significantly higher proteinase activity was found in blood serum of women with malignant tumors. PMID- 11386004 TI - Innovative clinical path slashes back surgery LOS. AB - Cooperative in Seattle overcame professional resistance and a corporate reorganization on its way to implementing protocols that drastically cut LOS for lumbar laminectomy. PMID- 11386005 TI - Involve nurses and patients to change physician behavior. AB - That's the advice from researchers at the University of Michigan who were surprised to learn that U-M physicians weren't exempt from the shortcomings documented among physicians nationally when treating AMI patients. They responded with aggressive internal and community-based initiatives to bring U-M physicians in line with guidelines from the American Academy of Cardiology and the American Heart Association. PMID- 11386006 TI - Florida initiative aims to slash unnecessary admissions due to 'catch-all' dehydration diagnosis. AB - When the peer review organization in Florida sounded an alarm about unnecessary medical admissions with the diagnosis of dehydration, a Tampa hospital decided to make the issue a priority. Although St. Joseph's Hospital was performing better than average, it still found ways to reduce the number of inappropriate admissions and improve related outcome measures. PMID- 11386007 TI - Isolation of the 5'-flanking region of genes by thermal asymmetric interlaced polymerase chain reaction. AB - BACKGROUND: Isolation of the 5'-flanking region of gene segments adjacent to known sequences is a tedious task. Thermal asymmetric interlaced (TAIL)-PCR developed in 1995 uses nested sequence-specific primers together with a shorter arbitrary degenerate primer so that the relative amplification efficiencies of specific and nonspecific products can be thermally controlled. In the present study, we modified the TAIL-PCR method to isolate the 5'-flanking region of two human genes, human natriuretic peptide receptor type A (NPRA) and type B (NPRB) genes. MATERIAL AND METHODS: We improved upon the original TAIL-PCR method in some aspects: three different series of eight AD primers were used simultaneously; a high denaturing temperature was used; and a Taq DNA polymerase developed for long and accurate PCR was used. RESULTS: We succeeded in isolating the 5'-flanking region of the NPRA and NPRB genes directly from human genomic DNA. CONCLUSION: Our TAIL-PCR method is applicable not only to be human genome but also to the genome of other life forms. PMID- 11386008 TI - Characterization of kappa and delta opioid receptors in isolated organs by using type/subtype selective agonists and antagonists. AB - BACKGROUND: The kappa and delta opioid receptors were characterized in longitudinal muscle strip of guinea-pig ileum (GPI, mu, kappa), mouse (MVD, delta, mu, kappa) and rabbit (LVD, kappa) vas deferens and rabbit ear artery (ART, delta, kappa) with particular attention to the presence of receptor subtypes. MATERIAL AND METHODS: For this purpose, type/subtype selective agonist and antagonist were used such as [D-Ala2, D-Leu5]-enkephalin (delta agonist) [D Pen2,5]-enkephalin (delta 1 agonist), deltorphin II (delta 2 agonist) naltriben (NTB, delta 2 antagonist), BOC-YPGFLT(OtBu) (delta antagonist) on the one hand and ethylketocyclazocine (EKC, kappa/(agonist), PD-117 302 (kappa 1 agonist), [Met5]-enkephalin-Arg6, Phe7 (ME-RF, delta/kappa 2/(agonist) and its amide ME-RF NH2, kappa 2/(agonist), naltrexone (NTX, mu > delta = kappa antagonist) and norbinaltorphimine (nBNI, kappa antagonist) on the other hand. RESULTS AND CONCLUSION: In MVD the Ke of NTB against different (agonists revealed no receptor type heterogeneity. In LVD the Ke of nBNI but not of NTX against EKC versus the ones against ME-RF and ME-RF-NH2 indicated inhomogenous (receptor population. The (receptor antagonist BOC-YPGFLT(OtBu) antagonized the action of ME-RF-NH2 in ART but not in MVD or LVD indicating a special receptor subtype in ART (kappa, possibly delta). PMID- 11386009 TI - Role of histamine in cardiac muscle function--an attempt at demonstrating the presence of intracellular histaminergic H2-type receptors. AB - BACKGROUND: Histamine is one of the biologically active substances that activate adenyl cyclase enzymatic system through H2 receptor. The aim of the study is demonstration of the presence of membranous histamine receptors in cardiomyocytes and determination of their role in function of the cardiac muscle cell. MATERIAL AND METHODS: The experiments were carried out on 20 rabbits divided into two experimental groups. Electric and mechanical activity of cardiac muscle trabecula was registered by glass microelectrodes' method in group I and by saccharose slit method in group II. RESULTS: In group I after administration of 2.5 microM x l-1 of histamine mean rhythm rate increased to 61 +/- 2.7 stimulations x min-1, contractile tone increased by 55.7 +/- 4.9% in relation to the control values, relaxation time shortened to 139.2 +/- 1.8 ms, while time to pike decreased to 229.6 +/- 2.5 ms. The following effects were observed after administration of 5 microM x l-1 of histamine: mean rhythm rate increased to 76.4 +/- 4.5 x min-1, amplitude of the potentials was unchanged, while their duration shortened to 227.7 +/- 3.2 ms, contractile tone increased by 122.4 +/- 12.9%, average. In group II contractile tone increased by mean 110%, time to pike decreased to 103 +/- 1.5 ms, while relaxation time--to 210.2 +/- 4.2 ms. Frequency of spontaneous stimulations as well as amplitude and duration of the action potential remained unchanged in all of the experiments. CONCLUSIONS: Positive chronotropic and inotropic action of histamine added to the extracellular fluid point to the presence of histaminergic receptor in rabbit's cardiac muscle. H2 histaminergic receptors are situated not only on the external surface of cellular membrane of rabbit's right heart atrium trabeculae, but also inside the cells. Experiments with histamine administration by the "cut end" method suggest that the role of intracellular H2 histaminergic receptors is associated with controlling the contractile processes of the cardiac muscle. PMID- 11386010 TI - The effect of spinal electrostimulation on the testicular structure in rabbit. AB - BACKGROUND: The main objective of this study was to create an experimental model of idiopathic scoliosis (i.s.), and to assess the effect of Lateral Electrical Surface Stimulation (LESS) on the organism both intra vitam and post mortem. The experiment made it possible to determine the extent to which LESS affects overall development of the organism, apart from its positive clinical effect in correcting i.s. in children and youth. An attempt is also made to explain the basis of systemic complications accompanying this method. MATERIAL AND METHODS: Studies were carried out on 10 white New Zealand male rabbits aged 3.5 months. They were divided into two groups, 5 animals in each group. The LESS group was stimulated using an SCOL-2 apparatus, 9 hours a day. The second group served as controls. After three months, the animals were sacrificed. Detailed macroscopic and microscopic examinations were performed on the rabbits' testicles. Scraps were collected immediately after the animal's death, from the free brim of the testis. The ultrastructure was examined with a TESLA BS-500 electron microscope. RESULTS: In the LESS group, histopathological examination of the testicles revealed considerable necrosis of the seminiferous epithelium, frequently coupled with peritubular fibrosis, atrophy of seminal tubules, and proliferation of Leydig cells. Ultrastructural examination revealed a multi-layered basal lamina, collagen appearing in the proper membrane of the seminiferous epithelium and blood vessels, lysis of supporting and sex cells of the tubular epithelium, mitochondrial damage, and the formation of myelin-like bodies in the round spermatids and the middle segment of the elongated spermatid tails. Lysis of the cytoplasm of Leydig cells was observed in the testes. CONCLUSION: Traditional electrostimulation induced regressive changes in the testes, in the form of necrosis of the seminiferous epithelium, atrophy of seminal tubules, and destruction of Leydig cells. PMID- 11386012 TI - GABA content and GAD activity in gastric cancer. AB - BACKGROUND: Several recent reports have suggested a connection between the GABA ergic system and neoplastic processes. It has been confirmed that both GABA content and GAD activity, are increased in neoplastic tissues in colon and breast cancer. In the light of the theory of dynamic balance between stimulating and inhibitory amino acids, disturbances in GABA metabolism may be an expression of the cell's defensive response to the neoplastic process. The aim of the present study was to evaluate GABA content and GAD activity in the tissue from gastric cancer and in the macroscopically unchanged wall of the stomach. MATERIAL AND METHODS: GABA content and GAD activity were evaluated in tissue material obtained from 34 patients operated for gastric cancer. GABA and GAD levels were determined in neoplastic tissue and surrounding unchanged tissue by spectrofluorometric method. RESULTS: The investigations revealed that the GABA content and GAD activity were significantly higher in the neoplastic tissue in comparison to the unchanged tissue of the stomach. CONCLUSION: The results obtained, together with reports from the literature, suggest the possibility of introducing GABA-ergic system agonists into adjuvant therapy in gastric cancer. PMID- 11386011 TI - Selected markers of oxidative stress in rats with active Heymann nephritis. AB - BACKGROUND: Reactive oxygen species play an important role in the pathogenesis of different glomerulopathies. The purpose of this study was to examine selected markers of oxidative stress and antioxidant defense in active Heymann nephritis (AHN), which is a model of human membranous nephropathy. MATERIAL AND METHODS: AHN was induced in female Wistar rats by i.p. injection of proximal tubule brush border antigen (Fx1A). The control animals received an equivolume injection of saline. The animals were sacrificed at 6, 9, 15, 17, 18, 19, 22 and 23 weeks after Fx1A administration. We assayed the plasma concentration of thiobarbituric acid-reactive substances (TBARS) and nitric oxide metabolites (nitrites + nitrates, NOx) as well as total plasma antioxidant capacity (ferric reducing ability of plasma, FRAP) and serum paraoxonase (PON) activity. RESULTS: Fx1A injected rats demonstrated a marked increase in proteinuria and impairment of renal excretory function evidenced by increased plasma creatinine and uric acid. Histologically, diffuse renal changes were seen, characterized by glomerular hypercellularity, increased glomerular size and narrowing of Bowman's space. Ultrastructural studies revealed diffuse fusion of foot processes and detachment of podocytes, subepithelial immune deposits in the glomerular capillary wall, thickening of the glomerular basement membrane, and local accumulation of lymphocytes and macrophages in glomeruli and interstitial cells. In the AHN group, TBARS increased significantly beginning in the 9th week, reaching maximum at the 23rd week (139.4% of time-matched control). The plasma concentration of NOx demonstrated a biphasic increase. The first peak (249.9% of control) was observed at the 9th week, followed by a decrease to normal between the 17th and 19th week. Then the NOx concentration increased to 211% of control at the 23rd week. FRAP began to increase in the 9th week and reached its maximum (134.9% of control) at the 15th week. After the 18th week FRAP returned progressively to control levels. PON activity was 17.0%, 19.1% and 21.3% lower in AHN than in the control group at the 19th, 22nd, and 23rd weeks, respectively. CONCLUSIONS: These results indicate that AHN is associated with oxidant-antioxidant imbalance, which may contribute to renal damage in this model of glomerulonephritis. PMID- 11386013 TI - Acute appendicitis: the role of enterotoxigenic strains of Bacteroides fragilis and Clostridium difficile. AB - BACKGROUND: The aim of this study was to investigate whether there is a relationship between enterotoxin-producing B. fragilis strains and toxigenic C. difficile strains and the pathogenesis of acute appendicitis. MATERIAL AND METHODS: Post-appendectomy tissues from 34 patients with histopathologically confirmed phlegmonous or gangrenous appendicitis were studied. RESULTS: Among 86 anaerobes isolated, the B. fragilis group was most frequently isolated: 34 B. fragilis strains were cultured from 21 post-appendectomy tissues. Two enterotoxin producing B. fragilis strains were found. Enterotoxin titers (1:10 and 1:160, respectively) were measured on HT29/C cells. The presence of the enterotoxin gene was confirmed by PCR in DNA extracted from both strains. Among 21 DNA samples isolated from those post-appendectomy tissues from which B. fragilis strains were cultured, the presence of the enterotoxin gene was confirmed in only one case (the corresponding B. fragilis strain enterotoxin titer was 1:160). A unique toxigenic C. difficile strain was also cultured from the tissue of an adult patient with gangrenous non-perforated appendicitis. The presence of toxin A and toxin B genes was confirmed by PCR in DNA extracted from the C. difficile strain, but these genes were not found in the DNA extracted from the corresponding tissue. CONCLUSION: The presence of enterotoxigenic B. fragilis and toxigenic C. difficile strains was shown in post-appendectomy tissue from patients with phlegmonous and gangrenous appendicitis, and the B. fragilis enterotoxin gene was detected directly in the corresponding tissue. Further investigations (including immunologic aspects) require to confirm the role of these toxins in pathogenesis of acute appendicitis. PMID- 11386014 TI - Influence of temporary ischemia on energetic functions of rat liver in early and late reperfusion period. AB - Energetic function impairment is one of the main reasons of liver ischemia reperfusion injury. The aim of the study was to estimate the influence of temporary rat liver ischemia on energetic functions of the organ in the early and late reperfusion period and to answer the question whether the reduction of destructive factors release in sinusoids in the early phase of reperfusion can prevent liver energetic function impairment. To perform the experiment 55 Wistar rats divided into 5 experimental groups were used. 60 min. partial ischemia of the liver was applied, with 30 min, 72 h and 5 days reperfusion period. Suppression of KC was performed by intravenous administration of GdCl3. In the group of animals subjected to liver ischemia a considerable decrease of reaction to SDH and LDH activity was observed in the reperfusion period. In animals with KC suppression the loss of reaction to SDH intensity in zones I of acini was insignificant and the decrease of reaction to LDH was more orderly and proceeded in a more regular manner. The value of AKBR fell significantly after 30 min. of reperfusion in both groups of animals subjected to ischemia, while the serum level of AspAT and LDH increased rapidly. Temporary ischemia of the rat liver induces a decrease of the organ energetic functions in the early and the reperfusion period. The elimination of Kupffer cell activity in the early reperfusion phase does not prevent liver energetic functions impairment, but reduces the time in which they return to initial values in remote observation. PMID- 11386015 TI - Benefits of the implementation of structured educational program in hypertension management. AB - BACKGROUND: The aim of the study was to evaluate the effect of patients' education on treatment compliance and blood pressure (BP) control, as well as lifestyle in hypertension. MATERIAL AND METHODS: Education with the structured program was performed in 89 patients with essential hypertension. The patients knowledge level was determined using a special questionnaire with 26 questions. The regularity of BP self-control and drug treatment were assessed before and after the educational course as well as after 6-months of follow-up. RESULTS: The knowledge level increased after education from 58 +/- 12% up to 92 +/- 11% (p < 0.05), the proportion of patients on regular medication from 34.4% to 74.2% (p < 0.05). Sixty (67.4%) of patients started to perform regular BP self-control. The improvement of knowledge level and treatment compliance was not accompanied by significant changes in lifestyle such as smoking cessation and body weight loss. CONCLUSIONS: The structured patients' education resulted in the increase of patients' knowledge, improvement of drug compliance and self-monitoring of BP. Modification of lifestyle appears to be more problematic point as it did not show any significant improvement. PMID- 11386016 TI - Neuropeptide Y in obese women during treatment with adrenergic modulation drugs. AB - BACKGROUND: The aim of the study is the assessment whether weight loss treatment with adrenergic modulation drugs modifies neuropeptide Y (NPY) plasma concentration in obese women. MATERIAL AND METHODS: 13 obese women (BMI 38.3 +/- 4.4) were tested before and subsequently 10 and 20 days after weight loss treatment. The treatment consisted of a very low caloric diet of 400 kcal (1670 kJ) daily combined with ephedrine with caffeine (E + C) or ephedrine with caffeine and yohimbine (E + C + Y) administered for 10 days using the cross-over method. The patients underwent physical examination, including heart rate and blood pressure measurements, spectral heart rate variability (HRV) at rest and after 3 minute handgrip and a 15 minute cycloergometer exercise at 75 W. All the above mentioned tests were carried out thrice in each patient. In 13 obese patients and in 6 control women plasma NPY concentrations were determined by a specific radioimmunoassay using rabbit anti-NPY antiserum and a standard synthetic porcine NPY (Peninsula Lab.). RESULTS: Plasma NPY concentrations were significantly lower in the obese persons compared with the control group. During weight loss treatment with adrenergic modulation drugs no changes in plasma NPY were found at rest and after physical exercise. Also no differences in HRV indices were observed. CONCLUSIONS: 1. Low plasma NPY concentration observed in obesity may be a contraregulatory factor that could prevent further weight increase. 2. Weight reduction treatment did not affect plasma NPY concentration and cardiovascular response to physical exercise. 3. The doses of adrenergic modulation drugs used in our study did not induce any serious side effects, and were so low that no change of plasma NPY concentration and cardiovascular responses were observed at rest. PMID- 11386018 TI - Increased plasma glutathione peroxidase activity in patients with acute myocardial infarction. AB - BACKGROUND: Aim of the study was to determine the concentration of selenium (Se) and the activity of glutathione peroxidase (GSH-Px) in patients with acute myocardial infarction (AMI) and to observe the behavior of these parameters during thrombolysis therapy. MATERIAL AND METHODS: The study comprised two groups of AMI patients and a control group. The first group consisted of 49 patients from whom blood samples were taken after admission to the intensive care unit and subsequently after 3, 7, 14 and 30 days of hospitalization. In the second group of patients (n = 18) blood was taken for measuring only the GSH-Px activity in plasma. In this group blood samples were collected after admission to the hospital, 6, 12, 24, 48 hours, 3, 7, 14 and 30 days later. Control group comprised of 58 healthy subjects. Se levels in whole blood and plasma were measured spectrofluorometrically with 2,3-diaminonaphthalene as a complexing reagent. GSH-Px activity in red cells and plasma was measured spectrofluorometrically with t-butyl hydroperoxide as substrate. RESULTS: In the first group of patients Se concentrations in whole blood and plasma as well as GSH-Px activities in red cells and plasma did not differ significantly from healthy subjects. Both Se levels and GSH-Px activities were stable during the entire period of the study. In the second group of patients, however, plasma GSH Px activity increased after admission and reached the highest value after 48 hours. This activity was significantly higher compared to healthy subjects (p < 0.004) and to the mean initial activity of this group (p < 0.02). In the later period the activity decreased to the values of healthy subjects. CONCLUSION: We suggest that the increased activity of GSH-Px in plasma of AMI patients is the response of the organism to the increased levels of reactive oxygen species produced during reperfusion and thrombolysis. PMID- 11386017 TI - Changes of plasma arginine-vasopressin level in patients with hyperthyroidism during treatment. AB - BACKGROUND: Hyperthyroidism is associated with several kinds of changes in the circulatory system, including alterations in blood volume. Arginine-vasopressin (AVP) is known to be one of the major factors regulating plasma volume. The present study was designed to evaluate the plasma AVP level in patients with hyperthyroidism during treatment. MATERIAL AND METHODS: AVP was measured under basal conditions and after stimulation with a low-sodium diet and upright position. Seventy-four patients with hyperthyroidism and 37 controls were investigated. Measurements were taken before treatment, two weeks after pharmacological treatment, and after attaining euthyroid status. The following indices were determined: AVP, total and free thyroxin and triiodothyronine, thyrotropin, sodium, potassium, hematocrit, arterial blood pressure, cardiac index, and total peripheral resistance index. Plasma osmolality and changes in plasma volume were calculated indirectly. RESULTS: Plasma AVP was higher in patients with hyperthyroidism before treatment. After normalization of thyroid status, the AVP level was similar to that of the controls. The application of a low-sodium diet and upright position resulted in a greater decrease in plasma volume than the controls. AVP correlated with thyroxin level and plasma osmolality in patients with hyperthyroidism. CONCLUSION: Enhanced AVP level in patients with hyperthyroidism is suggested to be the result of alterations in plasma volume and is relatively independent of changes in plasma osmolality. PMID- 11386019 TI - Various opportunistic infections and neoplasms in patients dying of AIDS in the last 12 years--report based on pathomorphological investigations. AB - BACKGROUND: The aim of the study was microscopic diagnostics of infections and neoplasms in patients who died of AIDS. MATERIAL AND METHODS: Post-mortem analyses were performed in the Hospital for Infectious Diseases in Warsaw. In total, 200 patients with AIDS who were treated and subsequently died were included in the analysis. The study group consisted of 175 men and 25 women. Patients' age ranged between 11 months and 64 years (mean age 43 years). RESULTS: The most frequent infection diagnosed in these subjects was cytomegalovirus, followed by bacterial, mycotic, tuberculotic infections and toxoplasmosis. As far as neoplasms are concerned, patients with AIDS suffered most frequently from malignant lymphoma, Kaposi's sarcoma and finally, from Hodgkin's disease. In most cases, many types of infections coexisted with one another or there was a co occurrence of the infection and neoplasm. CONCLUSIONS: The author suggests that the material presented in this paper is representative for Polish population. Pulmonary changes is the most often cause of the death of the patients with AIDS. PMID- 11386020 TI - Extra structurally abnormal chromosomes (ESACs)--presentation of 10 new cases. AB - BACKGROUND: The goal of the presented studies as a retrospective reliability assessment of classical banding cytogenetic studies and of prognosing epicrises in a group of 14 cases, affected with additional marker chromosomes. MATERIAL AND METHODS: Having collected the study material from peripheral blood, by means of trophoblast biopsy or amniocentesis, cytogenetic preparations were obtained, allowing for pre- or postnatal evaluation of the karyotype. A panel of auxiliary cytogenetic techniques accompanied the routine CTG protocol. RESULTS: In a group of 6875 persons with recommendations to pre- or postnatal cytogenetic diagnostics, 14 (0.2%) cases of ESACs were diagnosed. In 5 cases of DA/DAPI(+) inv dup (15) as observed. A presence of polymorphic interstitial RHG(+) band was found within the marker chromosome. The measured size of that band allowed associating it with either the presence or the absence of pathological signs. In 9 cases of ESACs, DA/DAPI(-), the application of banding techniques (NOR and CBG) allowed to discover bisatellite heterochromatic ESACs in 6 cases (2 non-mosaic and 4 mosaic). In three other mosaic and non-satellite cases of ESACs, a 'genetic inactivity' of the marker chromosome was observed in one case, while a 'genetic activity' was ascertained in two cases. The 'activity' of marker chromosomes was studied by means of replication banding techniques. CONCLUSIONS: At the time of the outburst of molecular techniques, still up-to-date is the use of classical banding techniques and of the replication techniques, allowing DNA replication kinetics studies at the level of single band. PMID- 11386022 TI - Frequency of the hemochromatosis C282Y and H63D mutations in a Polish population of Slavic origin. AB - BACKGROUND: Hereditary hemochromatosis is one of the most common monogenic diseases among Caucasians. The most frequent mutation causing hemochromatosis is C282Y in the HFE gene, the highest frequency of which has been observed in populations of Celtic origin. There are no studies providing information about the frequency of the C282Y and H63D mutations in the HFE gene in populations of Slavic origin. MATERIAL AND METHODS: We collected 871 healthy unrelated subjects in Poland to assess the relevant frequencies. Each subject was genotyped for the C282Y and H63D mutations using a PCR-based protocol. RESULTS: Among the analysed subjects 6.0% were CY heterozygotes, and only one person was YY homozygote. The observed frequency of the 282Y allele was 3.1%. The frequency of the D allele of the H63D polymorphism was 16.2%. There were 13 (1.5%) compound heterozygotes for C282Y and H63D found in the analysed group. CONCLUSION: The high frequency of the Y allele in this Polish population of non-Celtic origin may indicate an early introduction of this mutation through admixture with a strong positive selection. PMID- 11386021 TI - Research on some parameters of cellular immune response in soldiers undergoing basic training--preliminary report. AB - BACKGROUND: On the basis of available reports it can be stated that physical stress causes changes in distribution and activity of many components of the immune system. It is believed that psychophysical stress in soldiers can influence their immune system depressively and in consequence increase the risk of upper respiratory tract infections. Therefore, it was decided to conduct studies aimed at the estimation of the influence of military training on the some parameters of cellular immune response. MATERIAL AND METHODS: The study group consisted of 40 draft aged from 18 to 23 years. The research was conducted in the first 8 weeks of service, in the period of the most intense draft stress adaptation. The participants were divide into 2 groups, A and B respectively, 20 soldiers each. Group A derived from an assault unit. Their training induced strenuous physical stress. Group B derived from a support unit. Their training required less physical effort then one of group A. Performed examinations involved: lymphocyte percentage count, lymphocyte proliferative response to mitogen, CD69 antigen expression on T lymphocyte surface, delayed hypersensitivity reaction with CMI Multitest. All assessments were done twice at 8 weeks interval. RESULTS: After 8 weeks of training in the A group a statistically significant increase in the percentage of lymphocytes revealing antigens of the II Class Main Histocompatibility Complex (MHC) was found. In addition, in this group a statistically significant decrease in the value of lymphocyte stimulation index, a statistically significant increase in the percentage of cells revealing CD69 antigen expression after PHA stimulation were observed. During investigated period in the B group following statistically significant changes were found: an increase in the percentage of CD3+ and CD4+ cells, a decrease in the percentage of CD16+CD56+ and an increase in the CD4+ to CD8+ ratio. CONCLUSION: The obtained results show that military service conditions influence some parameters of the cellular immune response but do not result in the clinically significant suppression of the immune system. PMID- 11386023 TI - Pancytopenia in tuberous sclerosis. AB - A 46-year-old woman with edema and pancytopenia was referred for further evaluation. She was diagnosed as tuberous sclerosis with clinical manifestations such as facial adenoma sebaceous, ungual and periungual fibroma, subependymal nodules and renal angiomyolipoma. Her edema seemed due to hypercardiac function induced by massive anemia. X-ray revealed extraordinary thickening of the cortex of long bones of the extremities as well as patchy osteosclerotic findings in vertebra, suggesting that hematopoietic space was significantly reduced. Pancytopenia improved after splenectomy. Histological examination revealed several intrasplenic hemangiomas but its relationship to hypersplenism was not clear. It seemed that her massive pancytopenia was induced by a combination of hypersplenism and significant reduction in hematopoetic space. In tuberous sclerosis, various systemic complications sometimes induce severe hematological abnormalities. According to previous literatures, the present case of tuberous sclerosis manifested the most outstanding hematological complications. PMID- 11386024 TI - Coexistence of parathyroid carcinoma and non-medullary carcinoma of the thyroid. AB - A case of 35-year-old woman with parathyroid cancer is presented. Five years ago she underwent surgery for follicular thyroid cancer. Parathyroid cancer was evidenced by palpable, solid, irregularly shaped cervical tumor 5 cm in diameter. The patient had severe hyperparathyroidism confirmed by biochemical findings of hypercalcemia reaching 16 mg%, hypophosphatemia and hyperphosphatasemia. Serum parathormone level was 23-fold higher than the norm. These findings were accompanied by polyuria, polidypsia, symptoms of bone damage and renal calcification. After the surgery the patient's condition improved significantly despite persistent hyperparathyroidism. The level of parathormone decreased, but was still 11 times higher than the norm. Two months after the surgery she noticed a single node on her neck. The patient was re-operated for recurrence of parathyroid cancer. Serum parathormone level was then 6-8 times above the norm. Medical treatment with furosemide, calcitonin and biphosphonate resulted in normalization of calcemia and phosphatemia. Further management will aim at localization of foci of hyperactive parathyroid tissue in order to enable radical reoperation. The case is reported because of rare occurrence of parathyroid carcinoma as well as because the carcinoma occurred in a patient who previously had follicular thyroid cancer. There are no reports of coexistence of these two neoplasms in available literature. PMID- 11386025 TI - Iron deficiency anemia as the sole symptom of small intestine carcinoma. AB - BACKGROUND: The isolated longstanding hypochromic hyposideremic anemia can be a unique symptom of the jejunal tumor. CASE REPORT: The authors present a case of 43-year-old woman with small intestine cancer, which manifested as longstanding anemia, decreased serum iron and remained undiagnosed over a period of several years. Special attention has been paid to the problem of adequate diagnostic procedure for disclosing the latent small intestine tumor. PMID- 11386026 TI - Fast method for the estimation of heart valve leaflet viability. AB - BACKGROUND: The assessing of heart valve viability is critical for the preparing of viable and implantable biological value. Non-viable valves should be cross linked with various agents whereas viable tissues are suitable for cryoprotection. Cell viability assessment should be rapid, simple, easy and low cost. MATERIAL AND METHODS: A simple method, simultaneous fluorescent staining of both viable and damaged valve cells was described. The material was consisted of 2 groups: a) with ischemic time below 3 hours (5 swine valves), and b) with ischemic time over 3 hours (5 swine valves). The supravital staining of leaflet fragments was conducted with single solution of fluorescein diacetate (living cells staining) and propodium iodide (damaged cells staining). RESULTS: The simultaneous staining predominantly showed green fluorescence typical for living cells and few red damaged cells in the short ischemic time group, whereas the second group showed the predominance of the red stained damaged cells. CONCLUSIONS: The presented one-step supravital staining method is rapid and simple it allows differentiation of the viability of the valve cells, so it may be of value for the routine evaluation of cadaveric valve homografts. PMID- 11386027 TI - Application of FISH and Q-PCR techniques in breakpoint diagnostics in three cases of marker chromosomes derived from chromosome 15. AB - BACKGROUND: The goal of the study was a search for effective methods of diagnosing additional marker chromosomes. MATERIAL AND METHODS: Three cases of extra structurally abnormal chromosomes (ESACs) were diagnosed, the ESACs having been derived from chromosome 15 by cytogenetic techniques, the fluorescence in situ hybridisation (FISH) technique and the quantitative--polymerase chain reaction (Q-PCR). An application of a set of commercially available probes, specific for the 15q11.2-q12 regions (PWACR-Prader-Willi/Angelman Critical Region) allowed for a description of the breaking points. RESULTS: The presence of PWACR region was confirmed in one case and excluded in the other two. It was also attempted to apply the Q-PCR technique for a more accurate determination of the size of the region involved in chromosomal aberration, what would allow for a more reliable prognosing of the clinical outcome. In one of the patients, the breaking point was localized as distal to D15S144 locus, while it was proximal to D15S11 locus in the two remaining cases. CONCLUSIONS: The obtained results demonstrate a possibility of using the Q-PCR method in diagnosing unbalanced chromosome aberrations. PMID- 11386028 TI - Usefulness of the Poincare maps in detection of T-wave alternans in precordial leads of standard ECG--a comparison with the spectral method. AB - BACKGROUND: T-wave alternans (TWA) at microvolt level is considered as an important non-invasive risk factor for sudden death. Several methods are used to measure such repolarization variations, but each of them has some limitations. The purpose of our study is to assess the usefulness of Poincare maps, a method based on nonlinear dynamics theory, in detection of repolarization abnormalities. MATERIAL AND METHODS: In 30 postinfarction patients presence of TWA in precordial ECG leads was assessed by the spectral method (SM) and by the Poincare maps (PM). Quantitative measures of both methods: alternans voltage (AV) and alternans distance (AD) were compared using linear regression. RESULTS: Significant correlation between both measures (r = 0.92, p < 0.01) was found. The value of AD > or = 10 microV was accepted as significant for the presence of T-wave alternans. CONCLUSIONS: Poincare mapping seems to be a useful and simple method for detection of TWA. The alternans distance equal or greater than 10 microV can be considered as a level determinative for the presence of TWA. PMID- 11386029 TI - Allergic nasal polyps under scanning electron microscope. AB - The study material consisted of 18 patients (10 men, 8 women, aged 26-66 years), suffering from recurrent nasal polyps and bronchial asthma. Nine patients manifested intolerance to aspirin and non-steroid anti-inflammatory drugs (NSAIDs). The polyps removed during one of polypectomies were analysed with the help of scanning electron microscope (SEM) in order to evaluate the morphology of their surface. All the specimens displayed the increase in areas of cilium-free epithelium, covered with short microvilli and squamous epithelial cells. The arrangement of these changes was irregular and they were more pronounced in patients with intolerance to aspirin and NSAIDs. Apart from this, there were no other differences which would allow to differentiate in SEM investigation between polyps derived from patients with and without tolerance to NSAIDs. PMID- 11386030 TI - Effects of magnetic resonance imaging on polymorphonuclear neutrophil adhesion. AB - BACKGROUND: Limited research has been performed concerning the effects of MR imaging on the immune system. In this study the influence of MR imaging exposure on polymorphonuclear neutrophil (PMN) adhesion was evaluated. MATERIAL AND METHODS: In vivo and in vitro studies were performed in 10 patients undergoing an MR imaging procedure, PMN adhesion to a plastic surface, as well as the expression of adhesion molecules beta 2-integrins CD11b, CD18, and L-selectin on the surface of PMN were estimated. RESULTS: Exposure to MR imaging significantly increased adhesion of isolated PMNs to plastic surfaces. PMNs from blood samples obtained from patients undergoing MR imaging as well as from blood samples placed beside patients during MR imaging did not differ from controls in adhesion to plastic surfaces. Similarly, plasma from three tested samples did not change control PMN adhesion to plastic surface. Expression of beta 2-integrins (CD11, CD18) was significantly increased in samples left beside patients during MR imaging, while significantly decreased in samples obtained from patients after MR imaging exposure when compared to control samples. Expression of the surface adhesion molecule L-selectin on the surface of PMN decreased significantly in blood samples left beside patients during MR imaging. CONCLUSION: The results indicate that the PMN adhesion properties increase under the influence of MR imaging exposure. This phenomenon may be the result of direct stimulation of polymorphonuclear neutrophils by the exposure to MR imaging. PMID- 11386031 TI - Solving problems fluorine 19F with NMR spectroscopy. AB - Fluorine 19F is one of few 'lucky' elements found in living organisms, whose nucleus is characterised by non-zero spin, which makes it possible to detect fluorine and to analyse (and image) its chemical compounds with the help of Nuclear Magnetic Resonance (NMR). Fluorine 19F spectroscopy is not as popular as that of hydrogen 1H or phosphorus 31P, however, this method has recently been developed and applied (in medicine, too) more and more frequently, despite technical problems. This paper is a brief introduction to the techniques of investigating fluorine 19F with NMR spectroscopy, providing insight into the peculiarities and the difficulties associated with this method. It is also a short review of the achievements in NMR fluorine investigations in the fields of biology and medicine. PMID- 11386032 TI - Analysis of in vivo 1H MR spectra of normal brain tissue by means of second derivative method. AB - BACKGROUND: As follows from comparative studies conducted in a number of centres, meeting the repeatability and reliability conditions in MRS in vivo measurements depends mainly on system stability and operator-related external factors. The aim of the study was to check the usefulness of the automated fitting method offered by PeakFit 4.0 (by SPSS Inc., Chicago, USA). MATERIAL AND METHODS: Short echo time in vivo PRESS 1H MR spectra (2T, TE = 35 ms) of normal brain were fitted in the frequency domain using the second derivative method. In this approach local maxima and hidden peaks are found as local minima of spectrum second derivative. The Lorentzian robust minimization procedure using Levenburg-Marquardt non-linear fitting engine was applied. Spectral lines were approximated under the assumption of the mixed Lorentzian/Gaussian lineshapes. The same procedure was applied to 18 proton spectra recorded from frontal lobe white matter. RESULTS: The number of peaks found within the range of 0.74 + 4.2 ppm was 52 +/- 3 and their positions were almost the same. The fitted lines were assigned on the basis of the J pattern recalculated for the field strength of 2T and by comparing the chemical shifts with the shifts in single compound spectra. The ratios of main metabolites, such as NAA/Cr, Cho/Cr, Cho/NAA and mI/Cr, are in accordance with those obtained previously using the software supplied with the MR imager and the absolute concentrations of NAA, Cho, mI, Glc and Glu obtained from the fit agree with those reported in literature which confirms the usefulness of the second derivative method in routine analyses of 1H MR brain spectra. CONCLUSIONS: The automated fitting enables reasonable metabolite ratios and absolute concentrations to be obtained, however it should be kept in mind that the second derivative follows only the shape of the curve that represents the sum of all metabolites. PMID- 11386033 TI - The freeware AIDA interactive educational diabetes simulator- http://www.2aida.org--(1). A download survey for AIDA v4.0. AB - The purpose of this paper is to report a survey of 1,360 downloads of the AIDA interactive educational diabetes simulator. AIDA is a diabetes computer program which permits the interactive simulation of plasma insulin and blood glucose profiles for educational, demonstration and self-learning purposes. It has been made freely available, without charge, on the Internet as a non-commercial contribution to continuing diabetes education. Since its launch in 1996 over 74,000 visits have been logged at the AIDA Website--http://www.2aida.org--and over 20,000 copies of the AIDA program have been downloaded free-of-charge. This report documents a preliminary survey of downloaders of the software. The intended goals of the study were: (i) to establish the feasibility of using the Internet for auditing and surveying diabetes software users; (ii) to identify the proportion of patients with diabetes and their relatives who are actually making use of the program; and (iii) to establish certain technical details about downloaders' computer setups to facilitate the distribution of upgrades to the software. RESULTS: The Internet-based survey methodology was found to be robust and reliable. 1,360 responses were received over an 8 month period (from November 1999 to July 2000). During the corresponding period 3,821 actual downloads of the software were independently logged at the Website--giving a response rate to this survey of 35.6%. Responses were received from participants in 67 countries- although over half of these (n = 730, 54%) originated from the USA and UK. 762 responses (56%) were received from patients with diabetes and 184 (13.5%) from relatives of patients, with lesser numbers from doctors, students, diabetes educators, nurses, pharmacists, and other end users. Useful technical information about computers and operating systems being used were also obtained. This study has established the feasibility of using the Internet to survey, at no real cost, a large number of medical software downloaders/users. In addition it has yielded interesting data in terms of who are the main downloaders of the AIDA program, and has also provided technical (computer) information which has aided the recent release of a freeware upgrade to the software. PMID- 11386034 TI - The freeware AIDA interactive educational diabetes simulator- http://www.2aida.org--(2). Simulating glycosylated haemoglobin (HbA1c) levels in AIDA v4.3. AB - In 1996 an interactive educational diabetes simulator called AIDA was released without charge on the Internet as a non-commercial contribution to continuing diabetes education. Over the past 4+ years over 74,000 people have visited the AIDA Web pages at http://www.2aida.org and over 20,000 copies of the program have been downloaded from there free-of-charge. This article builds on the experience gained from the AIDA development, and the World Wide Web distribution of the software, and highlights some of the problems which users have reported with the program. An updated release of the software (AIDA v4.3) is described and the method applied for modelling glycosylated haemoglobin (HbA1c) levels within this new version of AIDA is documented. An overview is provided of the trialling and beta-testing of this latest release of the program, and the general concept of a 'virtual diabetic patient' that provides an electronic representation of a patient with diabetes--and which can be used for self learning/teaching/demonstration purposes--is highlighted. PMID- 11386035 TI - Is amiodarone the first or the last choice for the maintenance of sinus rhythm after successful conversion of atrial fibrillation? PMID- 11386037 TI - Pairing accountability with responsibility--the consequences of screening 'promotion'. AB - The chain of accountability, beginning with the inadequate information provision to women being invited for mammographic screening over the last decade, against a background of changing attitudes in women and increased understanding of DCIS, and the consequences and effectiveness of mammographic screening, is explored. In particular, an example is used of family repercussions arising from facile, unjustified extrapolation by insurance brokers of genetic risk by use of the 'breast cancer' label in a case of diagnosis of screen-detected ductal carcinoma in situ (DCIS) to seek to unjustly deny, restrict or withhold. The question posed is: where does the responsibility lie for such a serious, previously unadvised repercussion in a public health programme imposed on 'healthy' women, promoted to them in a coercive manner with unbalanced, inadequate information, and still not revised in the light of recent findings and GMC guidelines? PMID- 11386038 TI - Neurophysiology of the mental image. AB - Apart from perceptions, mental images are the most frequent experiences of the conscious mind. Mentation can be understood as a process which consists in the manipulation of recalled or imagined mental images. The scientific literature related to this process is relatively scant. This article reviews the relevant original and recent neurophysiological data in such a way as to enable the presentation of a synthetic theory of the neuronal mechanisms that form mental images. Recurrent axons and some reproductive efferent connections constitute the neural circuitry essential for imagery. These connections enable the reverberating, circular arousal of the upper levels of the hierarchical structure active during previous perceptions. The recalling of a mental image evokes an accompanying electromagnetic field. The investigation of its functional role is a challenge for contemporary neural scientists. PMID- 11386039 TI - Topical steroids--a new approach after 50 years. AB - This review article presents the current state of knowledge concerning the mode of action and side effects of corticosteroids, and efforts to eliminate iatrogenic clinical syndromes. PMID- 11386040 TI - Endogenous opioid peptides system in haemorrhagic shock--central cardiovascular regulation. AB - BACKGROUND: Haemorrhagic hypotension is accompanied by an increase in endogenous opioid system activity, however, its significance is not fully recognised. Therefore, the role of endogenous opioid peptides in the central cardiovascular regulation in haemorrhagic shock was reviewed. CONCLUSIONS: Complex central opioid mechanisms play an essential role in regulation of cardiovascular centre function in haemorrhagic hypotension since activation of mu and blockade of delta 1 opioid receptors inhibit the initiation of sympathoinhibitory phase of regulation, while activation of kappa receptors blocks the compensatory cardiovascular responses to haemorrhage. Evidences indicate that central nitrergic and glucocorticoid mechanisms are involved in endogenous opioids action. PMID- 11386041 TI - Biological heart valve--an alternative to mechanical valve. AB - Bioprosthetic heart valve, derived from human or animal tissues is a commercial, implantable valve characterised by good haemodynamic parameters, low haemolysis ratio and satisfactory durability. Until now, its wide-spread therapeutic use has been limited by the progressing leaflet calcification. The aim of the paper is to present the benefits, adverse reactions as well as problems associated with the improvement of this bioprosthesis. PMID- 11386042 TI - [Accreditation of health care organizations]. AB - The order No. 96-346 dated April 27, 1996 introduced in French Law, made it compulsory for all private and public health care organizations to be granted an official approval, the 'certificate of accredication'. This article aims at describing the main principles of a process which will surely profoundly modify quality management of health care in sanitary bodies. The first part is a reminder of the main concepts used in quality management which help us to understand and use the guidelines of accreditation distributed by the National French Agency for Accreditation and Assessement of Health Care (Agence Nationale pour l'Accreditation et l'Evaluation des etablissements de sante--ANAES). The second part of this article presents the procedure for granting the certificate of accreditation, especially its successive steps (the procedure launching, the self-evaluation phase, the visit of experts sent by ANAES, the report written by the experts and its aim). Beyond this procedure, a third part briefly underlines the main directions for developing efficient quality control and safety management of health care in sanitary bodies. PMID- 11386043 TI - [GPIIb-IIIa inhibitors]. AB - Therapy involving the use of anti-GPIIb-IIIa inhibitors has progressively evolved in recent years for patients undergoing percutaneous coronary intervention or with acute coronary syndromes. Patients receiving anti-GP IIb-IIIa therapy have a lower risk of death or myocardial infarction than those receiving the classic anti-agregant, aspirin, alone. Two classes of products have been used in clinic, the chimeric monoclonal antibody Fab fragment, c7E3 or abciximab (ReoPro), which has been the pioneer, and synthetic peptides or peptidomimetics such as eptifibatide (Integrilin) or tirofiban (Agrastat). Abciximab is a long-acting, high-affinity receptor blocker, whereas eptifibatide and tirofiban have much shorter biological half-lives. Another property that differentiates these compounds is that the peptides bind exclusively to GP IIb-IIIa whereas c7E3 also binds to alpha v beta 3, the vitronectin receptor. The potent inhibitory effect of these compounds increases the risk of bleeding. By carefully controlling the levels of heparin and by removing the sheath as early as possible, the hemorrhagic problems may be limited. Another potential complication is the rapid development of thrombocytopenia. The cause has yet to be found and for c7E3 no correlation with the development of HACA (human anti-chimeric antibodies) has been observed. Because of the chronic nature of coronary artery disease, evaluation of the readministration of c7E3 to the same patient two or even more times is under investigation. The first results do not show major problems. The best biological way to investigate the efficiency of anti-GPIIb-IIIa has to be determined. Interestingly, a new point-of-care test has been proposed, while monoclonal antibodies are available that differentiate between nonoccupied and occupied GPIIb-IIIa complexes. PMID- 11386044 TI - [Qualitative assessment of labile blood product prescription in Le Havre Hospital]. AB - This retrospective study assessed the conformity of prescriptions of labile blood products as well as the respect of legal requirements and recommendations concerning red blood cells specifications. This study was carried out in two phases one year apart, so as to be able to compare between them. It consisted in an analysis of the administrative and medical information present on the prescription forms and of the discrepancies between the prescribed product and that which was dispensed. Quality of prescriptions was overall better, but some points needed further improvement. The data moreover showed that some prescriptions of phenotyped and cross-matched red blood cells were incomplete. This did not improve over time. Out of 1000 prescriptions for red blood cells, 216 were altered by the transfusion service dispensing unit. A closer look at these changes showed that 49% were justified by statutory requirements or by local protocols. The discrepancies observed between the prescribed product and that which was dispensed were partly due to a poor understanding by the prescribers of statutory requirements. However they are mostly related to particular circumstances, such as emergencies. Furthermore, the prescribers expected the transfusion center to adapt the product in terms of the clinical context. PMID- 11386045 TI - [Impact of the exclusion of donors who have stayed in the United Kingdom on the residual risk of HIV transmission by blood transfusion]. AB - BACKGROUND: One of the measures aimed at reducing the risk of transmission of the agent responsible for the new variant of Creutzfeldt-Jakob disease was to exclude blood donors having stayed in the United Kingdom between 1980 and 1996. The objective of the study was to estimate the impact on the residual risk of HIV transmission of recruiting extra first-time donors to replace donors having stayed in the United Kingdom. METHODS: The residual risk of HIV transmission due to donations made during the window period was estimated in all donations made in France during the 3-year period 1996-1998 by a linear combination of residual risks in repeat donors and first-time donors. In repeat donors, the estimate is based on the incidence rate of HIV in this population and in first-time donors on the "detuned assay" method. Seven simulations of the impact on the residual risk were made using various percentages of donors which would be excluded (from -5% to -35%). RESULTS: In all donations made in France during the 1996-1998 period, the residual risk of HIV transmission was estimated at 0.70 per million donations, which represents five to six donations made during the window period. If all the donors who had stayed in the United Kingdom were excluded from the donation (35%) and replaced by first-time donors, the residual risk of HIV transmission would be increased from 0.70 to 0.86 per million donations. This increase of 24% would represent one or two extra cases of post-transfusion HIV infection over a 3-year period. CONCLUSION: The results of this study show that the exclusion of a large number of blood donors, replaced by first-time donors, would have a low but quantifiable impact on the residual risk of HIV transmission. This increase of risk was one of the factors that led to the decision of not excluding donors having stayed in the United Kingdom between 1980 and 1996. PMID- 11386046 TI - [Insisting on intravenous polyvalent immunoglobulin therapy in polymyositis in spite of the occurrence of sever hemolytic anemia]. AB - A 23-year-old female with polymyositis received high dose intravenous immunoglobulin (IVIg) therapy. The patient suffered severe hemolytic anemia after receiving first course of IVIg infusion. This adverse reaction was likely due to allohemaglutinin A and B and from or high molecular weight IgG complexes contained in the preparation. Though this effect was observed, the treatment was repeated six times. A mild hemolysis occurred following each IVIg, with no clinical consequence. Involvement of the saturation of macrophagic receptors might explain this partial destruction of erythrocytes. PMID- 11386047 TI - [Bacterial soil variety: evaluation of methods, possibilities, and prospects ]. AB - The paper presents a comparative description of the modern molecular genetic and routine culture techniques for assessing microbial diversity in soils and gives analysis for the different results obtained by these two groups of methods. The necessity of the collaboration of soil scientists, microbiologists, and molecular biologists in integrating different research methods for a proper assessment of soil microbial diversity is discussed. The paramount importance of soil as the source and reserve of biodiversity on the Earth is emphasized. PMID- 11386048 TI - [The role of putrescine in of oxidative stress defense genes expression regulation in Escherichia coli]. AB - The role of putrescine in the adaptive response of Escherichia coli grown aerobically in synthetic M9 medium with glucose to the H2O2-induced oxidative stress was studied. Under oxidative stress, the expression of the single-copy reporter gene fusions oxyR::lacZ and katG::lacZ was found to undergo biphasic changes, which were most pronounced in glucose-starved E. coli cells. The concentration-dependent activating effect of putrescine on the expression of the oxyR regulon genes was maximum when the oxyR gene was inhibited by high concentrations of hydrogen peroxide. PMID- 11386049 TI - [Saturated C21-C33 hydrocarbons are involved in the self-regulation of Pseudomonas fluorescens adhesion to a glass surface]. AB - One of the two putative groups of antiadhesions was identified in Pseudomonas fluorescens by the method of gas chromatography-mass spectrometry. A mixture of high-molecular unbranched hydrocarbons (HC) with a chain length from 21 to 33 carbon atoms reduced cell adhesion to a glass surface. These HC accumulated in the culture liquid to a total concentration of 10-15 micrograms/l; the concentrations of individual HC ranged from 0.1 to 3.0 micrograms/l. After the addition of individual HC to the bacterial culture, the number of cells attached to the glass surface decreased. This decrease in cell adhesion was due to the enhanced aggregation of the bacterial cells, which promoted mechanical (hydrodynamic) cell detachment from the surface. PMID- 11386050 TI - [Lithoautotrophic growth of the freshwater colorless sulfur bacterium Beggiatoa "leptomitiformis" D-402]. AB - The freshwater colorless sulfur bacterium Beggiatoa "leptomitiformis" D-402 was shown to be capable of lithoautotrophic growth in a batch culture under microaerobic conditions at O2 concentrations in the medium of no higher than 0.5 mg/l. The cell yield was maximum at a dissolved oxygen concentration of 0.15 mg/l. A high activity level of key enzymes of the Calvin cycle and of enzymes involved in dissimilatory oxidation of thiosulfate was recorded in the cells. The high rate of CO2 assimilation (112-139 nmol/(min mg protein)) and the cell yield (12 mg dry cells/mmol thiosulfate oxidized), 91-92% of which was accounted for by CO2 carbon, were close to those typical of autotrophic bacteria. Thiosulfate was oxidized almost completely to sulfate, and the fraction of elemental sulfur in the final products did not exceed 0.2-1.7% of the thiosulfate sulfur. The cell membrane fraction contained cytochromes (b + o) and two cytochromes c with M(r) of 23 and 26 kDa; the soluble fraction contained cytochrome c with M(r) of 12 kDa. PMID- 11386051 TI - [Metabolic characteristics of the Yarrowia lipolytica strain producing alpha ketoglutaric and citric acids from ethanol and the effect of [NH4+] and [O2] on yeast respiration and biosynthesis]. AB - The comparative studies performed in this work showed that overproduction of alpha-ketoglutaric acid (KGA) from ethanol by the mutant Yarrowia lipolytica strain I requires both a deficiency of thiamine and a relatively high concentration of ammonium ions in the medium, whereas CA overproduction requires an almost zero concentration of ammonium ions. The threshold value of the dissolved oxygen concentration in the medium, pO2, for CA overproduction is considerably higher than for KGA overproduction. The respiration rate of CA overproducing cells was 2-3.5 times higher than that of KGA-overproducing cells. The main terminal electron carrier functioning in the KGA-overproducing cells was cytochrome oxidase. In the CA-overproducing cells, the main terminal oxidase was presumably o-type cytochrome. PMID- 11386052 TI - [Effect of vitamin E and its functional analogs with different molecular structures on growth and lipids content of Pythium debaryanum]. AB - The effect of exogenously added vitamin E and its synthetic analogues (the hydrophilic form of vitamin E and chromans C13 and C1) on the growth, lipogenic activity, and the fatty acid composition of the eicosapolyenoic acid-synthesizing oomycete Pythium debaryanum was studied. The effect was found to depend on the molecular structure of particular compounds. For instance, vitamin E and chroman C13 stimulated fungal growth, whereas chroman C1 inhibited it. The hydrophilic form of vitamin E enhanced the lipogenic activity of the oomycete. The studied compounds, which possess antioxidant activity, did not exert any noticeable effect on the level and the degree of the unsaturation of fungal lipids. PMID- 11386053 TI - [Spatial peculiarities of rhizoplane colonization by microscopic fungi]. AB - Spatial peculiarities in the colonization of the tomato, cucumber, and barley rhizoplanes by microscopic fungi were studied. The apical zone of roots was colonized with a limited number of R strategists (the order Mucorales, Fusarium sp., Aspergillus niger, and Mycelia sterilia). The fungal population of the root hairs and the basal zone of roots was 2- to 3-fold more dense due to the prevalence of K strategists. Fusaria, Fusarium oxysporum in particular, colonized roots in earlier terms than the genera Trichoderma, Penicillium, Gliocladium, and others. The F. oxysporum population was at a maximum in the rhizoplane zone nearest the root tip. PMID- 11386054 TI - [Biogeochemical processes of methane cycle in the soils, swamps and lakes of Western Siberia]. AB - The biogeochemical processes of methane production and oxidation were studied in the upper horizons of tundra and taiga soils and of raised bogs and lake bottom sediments nearby the Tarkosalinsk gas field in western Siberia. Both in dry and water-logged soils, the total methane concentration (in soil particles and gaseous phase) was an order of magnitude higher than in the soil gaseous phase alone (22 and 1.1 nl/cm3, respectively). In bogs and lake bottom sediments, methane concentration was as high as 11 microliters/cm3. Acetate was the major precursor of the newly formed methane. The rate of aceticlastic methanogenesis reached 55 ng C/(cm3 day), whereas that of autotrophic methanogenesis was an order of magnitude lower. The most active methane production and oxidation were observed in bogs and lake sediments where the delta 13C values of CO2 were inversely related to the intensity of bacterial methane oxidation. Methane diffusing from bogs and lake bottom sediments showed delta 13C values ranging from -78 to -47@1000, whereas the delta 13C value of carbon dioxide ranged from 18 to -6@1000. In these ecosystems, methane emission comprised from 3 to 206 mg CH4/(m2 day). Conversely, the dry and water-logged soils of tundra and taiga took up atmospheric methane at a rate varying from 0.3 to 5.3 mg CH4/(m2 day). Methane consumption in soils was of biological rather than of adsorptive nature. This was confirmed by the radioisotopic method and chamber experiments, in which weighting of methane carbon was observed (the delta 13C value changed from -51 to 41@1000). PMID- 11386055 TI - [Phenotypic characteristics of Thiobacillus ferrooxidans strains]. AB - Phenotypic polymorphism of strains of Thiobacillus ferrooxidans isolated from various ecological niches was studied. The strains differed both in rates of growth and oxidation of Fe2+, S0, FeS2, and sulfide minerals contained in concentrate. Each strain, irrespective of its original environment, required a period of adaptation to a new substrate. Strains TFN-d, TFBk, TFO, and TFL-2, isolated from ores and concentrates rich in oxidized substrates, showed an equal adaptation pace (five culture transfers) but differed in their adaptation efficiency. Strain TFV-1, isolated from base ore and showing the lowest rates of growth and oxidation of all the substrates, required five culture transfers to adapt to S0 and FeS2 and seven culture transfers to adapt to the concentrate. It is concluded that the phenotypic properties of the strains correlate with their genotypic polymorphism and the environmental conditions under which their microevolution took place. PMID- 11386056 TI - [Effect of ecological factors on spore germination and the viability of the mycelial fragments of microscopic fungi]. AB - Spore germination and the viability of the mycelial fragments of the microscopic fungi Alternaria alternata, Penicillium spinulosum, and Mucor hiemalis were studied with respect to the action of some ecological factors: sucrose concentration (0, 0.2, 2, 10, and 100 g/l), temperature (4, 20, 25, and 30 degrees C), pH (3.5, 4.0, 5.0, 6.2, and 7.0), and cadmium concentration (0, 2, 10, and 100 mg/l). The spore germination rate and the viabilities of different mycelial fragments were found to reach their maxima at different values of the ecological factors studied. This finding suggests that the vegetative and asexual types of reproduction of microscopic fungi may have different ecological optima. PMID- 11386057 TI - [Molecular-genetic analysis of the yeast Komagataea (williopsis) pratensis populations from Caucasian and Tien Shan regions]. AB - The analysis of sixteen Komagataea (Williopsis) pratensis from the Caucasian and Tien Shan soils by the PCR, blot hybridization, and isoenzyme electrophoresis techniques showed that fifteen of them do belong to the species K. pratensis. The isolates from the two geographic areas differed in some physiological characteristics and in the PCR product profiles obtained with the microsatellite primers (CAC)5 and (GACA)4. PMID- 11386059 TI - [Coexistence of transgenic Escherichia coli strains and natural microorganisms in experimental aquatic microcosms]. AB - In experimental aquatic microcosms (AMCs), the population of the Escherichia coli strain Z905 harboring the recombinant plasmid pPHL7 (AprLux+) was found to gradually accumulate AMC-adapted cells, which retained the plasmid but differed from the original cells in some biochemical and physiological characteristics. Both the original and the AMC-adapted E. coli cells could coexist with the native AMC microflora for a year or longer. When introduced into AMCs together with native pseudomonads, the AMC-adapted E. coli Z905-33 (pPHL7) cells were more competitive than nonadapted cells. PMID- 11386058 TI - [Effect of mechanical stress on lon mutant strain of Escherichia coli K-12]. AB - It was found that, depending on their frequency, mechanical vibrations (MVs) can either stimulate (4 Hz) or inhibit (50 Hz) the growth and the division of the lon mutant of Escherichia coli K-12. Similar effects were observed when the MV treated nutrient medium was inoculated with untreated mutant cells. MVs enhanced the motility of mutant cells and the fragmentation of filament cells always present in the populations of lon mutants. PMID- 11386060 TI - [Proteolytic activity of lectins from the nitrogen-fixing bacterium Bacillus polymyxa]. AB - Two lectins (LI and LII) stripped from the surface of Bacillus polymyxa 1460 cells were found to possess proteolytic activity, which was associated with their hemagglutinating activity. The inhibition of the hemagglutinating activity of lectins by specific carbohydrate haptens changed their enzyme activities. The inhibition of hemagglutinating activity by glucuronic acid or fructose 1,6 diphosphate decreased the proteolytic activities of both lectins, whereas the blocking of this activity with D-glucosamine or D-galactosamine increased the proteolytic activity of LII. The molecules of the B. polymyxa lectins are suggested to contain two active centers responsible for hemagglutinating and proteolytic activities. PMID- 11386061 TI - [Processes of plant colonization by Methylobacterium strains and some bacterial properties ]. AB - The pink-pigmented facultative methylotrophic bacteria (PPFMB) of the genus Methylobacterium are indespensible inhabitants of the plant phyllosphere. Using maize Zea mays as a model, the ways of plant colonization by PPFMB and some properties of the latter that might be beneficial to plants were studied. A marked strain, Methylobacterium mesophilicum APR-8 (pULB113), was generated to facilitate the detection of the methylotrophic bacteria inoculated into the soil or applied to the maize leaves. Colonization of maize leaves by M. mesophilicum APR-8 (pULB113) occurred only after the bacteria were applied onto the leaf surface. In this case, the number of PPFMB cells on inoculated leaves increased with plant growth. During seed germination, no colonization of maize leaves with M. mesophilicum cells occurred immediately from the soil inoculated with the marked strain. Thus, under natural conditions, colonization of plant leaves with PPFMB seems to occur via soil particle transfer to the leaves by air. PPFMB monocultures were not antagonistic to phytopathogenic bacteria. However, mixed cultures of epiphytic bacteria containing Methylobacterium mesophilicum or M. extorquens did exhibit an antagonistic effect against the phytopathogenic bacteria studied (Xanthomonas camprestris, Pseudomonas syringae, Erwinia carotovora, Clavibacter michiganense, and Agrobacterium tumifaciens). Neither epiphytic and soil strains of Methylobacterium extorquens, M. organophillum, M. mesophilicum, and M. fujisawaense catalyzed ice nucleation. Hence, they cause no frost injury to plants. Thus, the results indicate that the strains of the genus Methylobacterium can protect plants against adverse environmental factors. PMID- 11386062 TI - [Role of the surface and extracellular substances of the phytopathogenic bacterium Xanthomonas campestris in its interactions with cabbage plants]. AB - Changes in some physiological and biochemical characteristics of cabbage (cv. Slava) seedling roots in response to inoculation with the phytopathogen Xanthomonas campestris and its surface and extracellular substances were evaluated. Seven days after the inoculation, the growth of the roots was slightly suppressed and they contained increased amounts of peroxidase. The effect of the lipopolysaccharides stripped from the cell surface or isolated from the culture liquid of X. campestris was similar to that of the whole cells of the phytopathogen. The bacterial lectin isolated from the cell surface material did not induce any defense response in cabbage plants but, presumably, could play a role in the contact interactions between bacteria and plants. PMID- 11386063 TI - [Study of extracellular structures of Agrobacterium involved in bacterial and plant interactions]. AB - Agrobacterial cells produced straight microfibrils not only when in contact with wheat seedling roots, but also when in contact with each other. After 2 h of incubation, agrobacterial cells were found to form aggregates, in which the cells were in contact either directly or through thick straight microfibrils (bridges) of an unknown composition. The majority of the microfibrils were susceptible to attack by cellulase, although some of them showed resistance to this enzyme. Like the wild-type flagellated agrobacteria, their bald mutants produced long straight microfibrils. The cells surface structures of agrobacteria were examined by labeling them immunocytochemically with colloidal gold conjugated antibodies against O-specific lipopolysaccharides, Vir proteins, and cellulase. Agrobacterial cells treated with acetosyringone and brought into contact were found to contain subpolar and polar cell surface structures. Antibodies against the VirB2 protein were able to interact with a tuft of thin microfibrils located on one pole of the agrobacterial cell, whose vir genes were induced by acetosyringone, but were unable to interact with the surface structures of the agrobacterial cells aggregated in liquid medium in the absence of wheat seedlings. PMID- 11386064 TI - [Tolerance of marine proteobacteria Pseudoalteromonas and Alteromonas to heavy metals compounds]. PMID- 11386065 TI - [International symposium "Modern problems of biochemistry and biotechnology of microorganisms]. PMID- 11386067 TI - Learning to heal, one patient at a time. PMID- 11386066 TI - These doctor's mission: keeping drug costs down. PMID- 11386068 TI - What made 50 people so sick? PMID- 11386069 TI - How to avoid medication errors. PMID- 11386070 TI - Can you speak your mind and keep your job? PMID- 11386072 TI - E-mail etiquette. PMID- 11386071 TI - $739,000 in debt--and wanting to start a family. PMID- 11386073 TI - Moonlighting inside the big house. PMID- 11386074 TI - Dad and I--and how we've helped each other. PMID- 11386075 TI - Are you collecting illegal dollars for "incident to" services? PMID- 11386076 TI - An algorithm for targeted convergence of Euler or Newton iterations. AB - The concept of multistationarity has become essential for understanding cell differentiation. For this reason theoretical biologists have more and more frequently to determine the steady values, often multiple, of systems of non linear differential equations. It is well known that iteration processes of current use converge or not towards a fixed point depending on the absolute value of the slope of the iteration function in the vicinity of the considered fixed point. A number of methods have been developed to obtain or accelerate convergence. As biologists, we do not pretend to review these works. Rather, we propose here a simple algorithm which permits to converge at will towards a chosen type of steady state. Others and we have used this procedure extensively for years for the analysis of complex biological systems. A compact program (using Mathematica) is available. PMID- 11386077 TI - Binding energy and the information content of some elementary biological processes. AB - Protein interactions within a multimolecular complex can result in information and energy transfer between proteins. This can lead in turn to the emergence of novel functions of some proteins of the complex. Various examples of this situation can be found in the scientific literature. This is probably the case for prion protein, chloroplast phosphoribulokinase bound to glyceraldehyde phosphate dehydrogenase, Ras system, and pancreatic lipase bound to biomembranes, to cite but a few. Any enzyme reaction, or enzyme reaction network, carries Shannon entropy and information. On contrary to genome entropy, the entropy of enzyme reactions and metabolic sequences is sensitive to 'external' signals, such as substrate, effector and proton concentrations. Complex structural organization of the cell is associated with a higher entropy content, and one can calculate the gain of entropy and information due to integration and complexity. One may conclude from this brief analysis that the informational content of a living cell is much larger than that of its genome. PMID- 11386078 TI - A comprehensive phylogeny of the Phaeophyceae based on nrDNA sequences resolves the earliest divergences. AB - The present study is the first comprehensive phylogeny of the class of the Phaeophyceae. For 67 species representing all orders of the class, the sequences of the 3'-end of the small and the 5'-end of the large subunit nrRNA genes were aligned and analysed. A further analysis based on sequences of the 3'-end of the small and of the complete sequences of the 28S gene of the large subunits was also performed, but for only eight taxa. In both analyses, Tribonema marinum (Xanthophyceae) was used as outgroup. The analyses showed the Dictyotales as diverging first, followed by the Sphacelariales, then the Syringodermatales. Most of the orders currently accepted were confirmed as monophyletic groups but the Laminariales and Tilopteridales remained polyphyletic. The relationships of the remaining orders to each other were not resolved with the present data set. Ascoseira, included for the first time in a molecular study, appeared as a separate lineage without any clear relationship with other algae possessing conceptacles (Splachnidium and Fucales). Algae with stellate plastids, never studied in a global context, were polyphyletic; this result is consistent with their plastid ultrastructure and is discussed in detail. As further result of the present study, the South African genus Bifurcariopsis appeared as the sister taxon of the North Atlantic genus Himanthalia, and Xiphophora appeared as the sister taxon of Hormosira rather than as a member of the Fucaceae; the taxonomic position of these genera is discussed. PMID- 11386079 TI - [Rapid determination of sex in Myocastor coypus embryos in the first stage of gestation]. AB - The early knowledge of the sex may be crucial for the understanding of many features of ecological and evolutive biology, including offspring sex-ratio adjustment and evolution of breeding systems. In coypu (Myocastor coypus), significant variation in birth sex-ratios can be observed and selective abortion of entire litters is one of the cited mechanisms. In order to determine the sex of coypu embryos in the earlier stages of gestation (second week), we developed a molecular technique based on PCR amplification of a region of the Sry gene. These method used the combination of two sets of primers: one specific of the Y chromosome; the other one, autosomal, is a positive control for amplification. Because of the direct amplification of embryo lysate without DNA extraction, the present sexing technique is rapid, relatively simple and inexpensive, and presents numerous advantages for the study at population scale. PMID- 11386080 TI - [Daltonism and the genetics of aging]. AB - In order to test whether mutations giving rise to color vision deficiencies are more frequently inherited from older fathers, an exhaustive screening of births in the Namur region has allowed to isolate a sample of 225 descending sons of maternal grandfathers who were older than 45 years at their daughter's birth. The incidence of color vision defects was compared between this set of cases and three control groups totalling 959 boys from independent families. While these comparisons were not conclusive, we propose new hypotheses concerning the population dynamics of color vision deficiencies. Neomutations in X-linked pigment genes may be a marker of the overall genetic load borne by the X chromosome. Selection against such loaded X chromosomes may occur in the second generation, either in the course of embryogenesis, or during female gametogenesis. The future assessment of these novel hypotheses relies on the arbitration of molecular genetics. PMID- 11386081 TI - Purification of several pectin methyltransferases from cell suspension cultures of flax (Linum usitatissimum L.). AB - Three pectin methyltransferases (PMT5, PMT7, PMT18; EC 2.1.1.6.x) were solubilized from the endo-membrane complex of flax cells, with 0.05% Triton X 100. After a 3 step-chromatography procedure, PMT7 and PMT5 were purified to apparent homogeneity. PMT5 and PMT7 differed regarding their optimum pH (5 or 7), the methyl acceptor (low or highly methylesterified pectin), their focusing pH range (6-7 or 8-9) and relative molecular mass (40 +/- 5 or 110 +/- 10 kDa). SDS PAGE of PMT5 and PMT7 did not reveal bands at 40 or 110 kDa but only a silver stained band of about 18 kDa. Two independent methods (photo labelling and enzymatic activity) showed that this silverstained band corresponded to a methyltransferase with affinity for pectins. This polypeptide was of the same size as the enzyme designed PMT18 (18 +/- 3 kDa; pl 4-4.5) recovered during size exclusion chromatography of either PMT7 or PMT5, suggesting that PMT18 bears the catalytic site of PMT5 and PMT7. PMID- 11386082 TI - [Determination of the state of emaciation of eel (Anguilla Anguilla) in migration in coastal and estuary zones]. AB - The objective of this work is to characterise the variations of the quality of the glass eels recruitment during a season of migration, from November till March and before and after the transition from marine to continental environment. The emaciation state measured from the ratio DNA/Dry weight and percentage of body water is analysed from a sampling of glass-eels collected at sea and in estuary, in the South of the Bay of Biscay during the migration period 1999/2000. The length and the mass are taken into account, they decrease during the season while the individuals are more and more pigmented. However, our work shows that groups of glass eels arriving on the coasts of the Bay of Biscay are homogeneous from the point of view of their energetic and hydride reserves with a high individual variability within these groups. Also no significant difference in losses of water and energetic reserve were observed among the individuals migrating at the beginning and during tide stream. Fluctuations of the biochemical indication and the percentage of water between the glass eels caught in sea and in estuary show that the transition between maritime and continental environment is not a mobilising event of a lot of energy for glass eels. Differences were only observed in December. PMID- 11386083 TI - Carbon isotope discrimination and mineral composition of three organs in durum wheat genotypes grown under Mediterranean conditions. AB - Carbon isotope discrimination (delta) has been proposed as a good criterion for transpiration efficiency and grain yield improvement. Its measurement, however, remains very expensive. Ash content (ma) has been proposed as an alternative criterion for delta in bread wheat and barley. The aims of this study were (i) to analyse the relationships between delta and mineral composition in different durum wheat plant parts and (ii) to compare the variation of these traits between landraces and improved varieties from different geographic origins. For this purpose, delta, ma, and composition in four minerals (K, Mg, P and Si) were assessed in flag leaves and awns at anthesis, and in mature grains of ten durum wheat genotypes grown under rainfed Mediterranean conditions. The three plant parts differed significantly for the measured traits. Significant correlations were noted between delta and ma in the flag leaf and in the grain. Silicon content in flag leaves and potassium content in awns were also positively related to delta of the considered plant part. The coefficient of correlation between delta and ma was generally higher than that observed between individual mineral content and delta, suggesting that ma is the better alternative criterion for delta. In addition, grain yield was related to grain delta and both ma and potassium content in awns. Harvest index was correlated with delta and ma of grain and flag leaf. These results emphasised that ma values in flag leaf and grain represent the efficiency of carbon partitioning to the grain. Improved varieties showed higher delta and ma values than landraces. Differences between Middle-East and West Mediterranean genotypes for the measured traits were also presented and discussed. PMID- 11386084 TI - The proliferation of human T lymphoblastic cells induced by 5-HT1B receptors activation is regulated by 5-HT-moduline. AB - Serotonin (5-hydroxytryptamine, 5-HT) is a well-known neurotransmitter and immunomodulator, which has been reported to affect the function of cells in the immune system. The purpose of the herein reported experiments was to investigate whether serotonin could regulate the proliferation of a human T lymphoblastic leukemia cell line (CCRF-CEM cells) and to characterize the 5-HT receptor(s) involved in this phenomenon using a pharmacological approach. The herein presented results show that serotonin alone stimulated the proliferation of CCRF CEM cells and that this effect could be mimicked by two 5-HT1B/1D receptor agonists (L-694,247 and GR 46611). Serotonin- or L-694,247-induced increase in cell proliferation was inhibited by a selective 5-HT1B receptor antagonist, SB 224289. A recently identified endogenous tetrapeptide, 5-HT-moduline (Leu-Ser-Ala Leu, LSAL), which specifically antagonizes 5-HT1B/1D receptor activity, was also shown to reverse the stimulating action of L-694,247 on T cell proliferation. Taken together, these results establish the existence of a direct serotonergic control of the T cell proliferation mediated through h5-HT1B receptors. In addition, these results are in favour of an immunomodulatory role of 5-HT moduline. PMID- 11386085 TI - [The Lateglacial sequence of the Etelles (northern French Alps): climate instability and vegetation dynamics]. AB - A detailed pollen analysis has been conducted in a new sedimentary sequence taken at Les Etelles (700 m), in the hill vegetation belt in the Chaine des Hurtieres (Northern French Alps). The Lateglacial history of the vegetation has been supported by 10 AMS datings. Variations in relative abundance of main pollen taxa were compared with variations in [symbol: see text] 18O in Swiss lacustrine sediments and with the GRIP ice-core record. Those highlight how the vegetation reacted to minor climatic oscillations. The Lateglacial spread of Quercus, Alnus and Corylus in the region is confirmed, thus allowing a new interpretation of some Lateglacial sedimentary sequences at an adjacent region (Bas-Dauphine). PMID- 11386086 TI - [Comparison of optical methods for estimating the opening of the canopy and leaf area in leafy forests]. AB - Based on inversion of gap fraction data (Poisson model of foliage distribution), three optical methods using the Demon, the Plant Canopy Analyzer LAI-2000 (PCA) and hemispherical photographs, have been compared to estimate canopy openness (CO) and leaf area index (LAI) in a mature, neutrophil, oak-beech-hornbeam forest on mull in eastern France. Mean CO over the whole hemisphere was similar for PCA (7.9%) and hemispherical photographs (8.0%). The needle method, a vertical point quadrat method, applied to the litter after leaf fall has served as a reference to LAI (4.7). The Demon provided the estimate (4.9) closest to the reference value. The PCA and hemispherical photographs underestimate mean LAI by 30% (3.3) and 19% (3.8), respectively, if used without correction. Based on fish-eye sensors, LAI estimates can be improved if 3 annuli (4.2) or 2 annuli (4.5) are used in place of 5 with the PCA, or by means of logarithmic averaging of gap fractions over azimuth at an appropriate angular resolution (180 degrees: 4.6, or 120 degrees: 5.2) with hemispherical photographs. Not taking into account azimuthal variation in gap fraction distribution generates a more important error than the error induced by light scattering near horizon. In order to improve LAI estimates, an original iterative procedure is presented, which allows the simultaneous calculation of LAI over a broad range of angular azimuthal resolutions. PMID- 11386087 TI - Basal teleosts and the question of elopomorph monophyly. Morphological and molecular approaches. AB - The methodology used by previous authors to resolve the relationships of the elopomorphan taxa is criticized. The morphological characters that have been proposed to support the monophyly of the Elopomorpha are reviewed and it is shown that most of them are weak. A new hypothesis of relationships is proposed on the basis of nucleotidic sequences of ribosomal RNA 18S, 16S and 12S. In order to really test all the possible relationships, the monophyly of the Elopomorpha was not considered a priori. The tree was rooted on Amia calva and Lepisosteus osseus and the ingroup taxa sampling was subsequently increased. The obtained topology shows that the Elopomorpha is a non-monophyletic taxon. Elopiforms, anguilliforms, albuliforms and notacanthiforms are considered here as four monophyletic, incertae sedis taxa among basal teleosts. PMID- 11386088 TI - [Airway management--is there more than one "Gold Standard"?]. PMID- 11386089 TI - [Treatment of acute renal failure--concepts and controversies. 2. Extracorporeal renal replacement and peritoneal dialysis]. AB - Therapy of prolonged acute renal failure regularly requires a renal replacement therapy. This can be achieved by different extracorporal renal replacement therapies (ERRT) or by peritoneal dialysis. ERRT are classified according to the physical principle underlying toxin elimination as hemodialysis (diffusion) and hemofiltration (convection). Another classification refers to intermittent or continuous application modes. Biocompatibility of membranes is judged according to their activation of the complement system. Prospective randomized studies did not consolidate the assumptions about the benefit of particular modalities proposed on theoretical foundations. Mortality, duration and complication rates of acute renal failure are not significantly decreased by use of biocompatible membranes. Continuous modalities are not generally preferable but optimize treatment in hemodynamically unstable patients, in whom they endorse fluid balancing and maintenance of sufficient arterial blood pressure. The use of demanding hemofiltration techniques for cytokine removal should be limited to clinical studies. The effects of ERRT-"intensity" and the best timing for initiation of ERRT have not been evaluated sufficiently. The choice of the ERRT modality is subject to clinical judgement (criterion: hemodynamic situation), practical aspects (criteria: availability of equipment and handling experience), and costs. Prior to their general use new and expensive technical modalities and membrane types should be thoroughly evaluated in studies with regard to outcome related aspects such as patient survival and preservation of renal function. PMID- 11386090 TI - [Circulatory mechanics in intra- and extraperitoneal CO2 insufflation in minimally invasive procedures. The effects of a low pressure system]. AB - Several studies have addressed the cardiovascular effects of intraperitoneal carbon dioxide insufflation and increased intraabdominal pressure. The pathophysiology of this intervention is complex. Reported results apparently differ depending on which patients are studied and are affected by blood volume and/or positioning. With the Starling resistor concept of abdominal venous return in which, analogous to pulmonary vascular zones, flow through the inferior vena cava is considered a function of the pressure difference between upstream venous and either abdominal pressure or downstream intrathoracic caval vein pressure, different results reported in literature can be reconciled. PMID- 11386091 TI - [The ProSeal-LM, a new variant of the laryngeal mask: description and first clinical results]. AB - The ProSeal-LMA is a modification of the laryngeal mask airway (LMA) invented by AIJ Brain. It differs from the known models in three major respects: An additional lumen extending into the tip of the cuff allows insertion of a gastric tube. A pocket immediately behind the cuff accommodates a finger or a siliconated metal introducer and thus facilitates insertion. An additional cuff at the dorsal aspect of the mask pushes the mask into a ventral direction and thus improves the seal around the glottic opening. Our experience after 8 months of clinical use and first data from an international multi-center-study (24 ProSeal-LMA/24 Standard-LMA) are reported. With a ProSeal-LMA insertion of a gastric tube is much easier and more reliable than with a Standard-LMA (24/24 patients vs. 2/24 patients). Judged from visibility of the oesophagus during fibre-endoscopy the position of the ProSeal-LMA is superior to that of the Standard-LMA. Both are equally well tolerated and handled. Most remarkable is a better seal for the ProSeal-LMA (pressure for a leak of 3 L/min: 31 mbar compared to 20 mbar (mean) for the Standard-LMA). Therefore the ProSeal-LMA seems suitable for anaesthesia procedures with IPPV, which up to now are a domain of endotracheal intubation. PMID- 11386092 TI - [Ropivacaine 0.2% versus bupivacaine 0.125% plus sufentanil for continuous peridural analgesia following extended abdominal operations]. AB - OBJECTIVE: An improved opioid-free analgesia concept using thoracic epidural anlgesia with a low concentrated local anaesthetic alone could be of practical interest for hospitals which are unable to guarantee continuous monitoring on the wards. METHODS: We have compared in a prospective, randomized, double-blind study, the analgesic effects and side effects of ropivacaine 0.2% (Group 1) with bupivacaine 0.125% plus 0.5 microgram ml-1 sufentanil (Group 2) via a thoracic epidural catheter during the first 24 hours after major abdominal surgery in 28 gynaecological tumour patients. Assessments were performed 8, 12, 16, 20 and 24 hours after surgery. RESULTS: No differences were seen in duration of surgery, intraoperative opioid and supplemental analgesic requirements. As a main result VAS values for pain during mobilisation were higher in Group 1 when compared with Group 2 (p < 0.01) after 20 and 24 hours of continuous epidural infusion (10 ml h 1) whereas VAS values at rest were comparable (p < 0.1) at all measurepoints. Intensity of motor block and side effects did not differ between groups. CONCLUSION: The present study shows that continuous epidural infusion of 0.2% ropivacaine alone provides comparable analgesia at rest, but less profound pain relief during mobilisation within the first 24 hours after major abdominal surgery than bupivacaine 0.125% with sufentanil. PMID- 11386093 TI - [Perioperative infusion therapy]. PMID- 11386094 TI - [Cerebral pontine myelinolysis--a grave treatment outcome?]. PMID- 11386095 TI - [Amniotic fluid embolism as an obstetric emergency--description of the syndrome and a case with fatal outcome]. AB - Amnotic fluid embolism is an incompletely understood obstetric emergency presenting with the acute onset of hypoxia, hypotension and severe coagulopathy. Up to 86% of the cases are fatal, maternal death occurs within the first hour of symptom onset in 50% of the patients. This report describes the case of a 29-year old, HIV-positive patient undergoing elective cesarean section who died despite immediate onset of cardiopulmonary resuscitation. The epidemiology of amniotic fluid embolism, clinical presentation, differential diagnosis, pathology and possible treatment are discussed. PMID- 11386096 TI - [Questions and answers in AINS]. PMID- 11386097 TI - [Late outcome after implantation of the PFC modular knee system]. AB - AIM: The objective of the current, prospective study was the evaluation of long term results after total knee arthroplasty with the PFC-Modular-knee system. METHOD: A total of 514 total knee arthroplasties were implanted between 1991 and 1994 in 248 female and 55 male patients. Their average age amounted to 62.4 years and 449 of them (87.4%) were re-investigated within the first half-year of 2000. Clinical and radiological data were evaluated according the recommendations of the American Knee Society. RESULTS: Late complications were observed in 6.5% of the cases. They consisted of prosthetic loosening (13 knees), patella problems (11 knees), instability (2 knees), periprosthetic fractures (2 knees), and one prosthesis fracture, 18 patients complained about a therapy-resistant, chronic pain syndrome. These findings lead to revision arthroplasty in 15 cases (3.3%). Preoperative American Knee Society Score values were on average 33, 35 an 17 points and improved to mean values of about 85, 79 and 5 points. Radiological signs of prosthetic loosening were observed in 12 cases (2.7%). CONCLUSION: Long term results after total knee arthroplasty with the PFC-Modular-knee system showed a significant improvement in the patients' complaints and clinical findings at an average of 7.8 years postoperatively. Complication and revision rates of 6.5% and 3.3%, respectively, were low when compared with the international literature and--apart from this--decreased in the course of the observation time. PMID- 11386098 TI - [Navigation in knee endoprosthesis implantation--preliminary experiences and prospective comparative study with conventional implantation technique]. AB - OBJECTIVES: The efficiency of a computer-integrated instrumentation system in knee arthroplasty was evaluated and compared with a conventional instrumentation system. BACKGROUND: The OrthoPilot System defines the individual axis of the leg by means of an intraoperative kinematic analysis. LED's mounted on rigid bodies and screwed to the pelvis, femur and tibia are localized by a 3D infrared camera which is linked to a UNIX work station. The integrated calculation program leads to definition of the centres of hip, knee and ankle. Thereafter, LED-equipped alignment instruments allow definition of the femoral and tibial main resection planes. METHOD: The first sixty cases were included in the study. In addition, thirty cases each were entered into an OrthoPilot group and in a similar conventional control group. The navigated cohort consists of cases one to thirty, thus enclosing the "learning curve". RESULTS: Leg axes and femoral and tibial angles were assessed radiographically at the three-months postoperative control. Radiological measurements of the OrthoPilot group were clearly superior to those of the manual group. The differences, however, were not statistically different in the parameters "mechanical axis", "femoral axis lat." and "tibial axis ap.". With regard to the parameter "tibial axis lat." a significant difference in favour of the navigation system was observed. The measurements of "femoral axis ap." were insignificantly better in the manual group. In general, a slight tendency towards valgus positioning of the femoral components when using the navigation system has to be discussed. Complications influencing the clinical outcome did not occur. Additional time for navigation is calculated in a range of ten to fifteen minutes. CONCLUSIONS: The OrthoPilot system clearly facilitates proper alignment of endoprosthetic components in femur and tibia. Generally, the obtained values representing endoprosthetic alignment are superior to conventional technique. Marked deviations from ideal alignment can almost be avoided by means of the navigation system. During the learning curve the OrthoPilot system gained in reliability and reproducability. PMID- 11386099 TI - [Navigated implantation of total knee endoprostheses--a comparative study with conventional instrumentation]. AB - OBJECTIVE: A navigation system should improve the quality of a total knee prosthesis implantation in comparison to the classical, surgeon-controlled operative technique. METHODS: The authors have implanted 40 knee total prostheses with an optical infrared navigation system (Orthopilot AESCULAP, Tuttlingen- group A). The quality of implantation was studied on postoperative long leg AP and lateral X-rays, and compared to a control group of 40 computer-paired total knee prostheses o the same model (Search Prosthesis, AESCULAP, Tuttlingen) implanted with a classical, surgeon-controlled technique (group B). RESULTS: An optimal mechanical femorotibial angle (3 degrees valgus to 3 degrees varus) was obtained by 33 cases in group A and 31 cases in group B (p > 0.05). Better results were seen for the coronal and sagittal orientation of both tibial and femoral components in group A. Globally, 26 cases of the group A and 12 cases of the group B were implanted in an optimal manner for all studied criteria (p < 0.01). CONCLUSIONS: The used navigation system allows a significant improvement of the quality of implantation of a knee total prosthesis in comparison to a classical, surgeon-controlled instrumentation. Long-term outcome could be consequently improved. PMID- 11386100 TI - [Knee endoprosthesis implantation in hemophiliac arthropathy: results, problems and complications]. AB - AIM OF STUDY: In the final stages of haemophilic arthropathy of the knee joint, the preservation of walking ability is only possible by joint replacement. Fibrotic ancylosis and severe deformities, being mostly bilaterally, make the joint reconstruction difficult and impair the results. The purpose of this study was to evaluate the results of total knee replacement (TKR) in haemophilia. METHODS: From 1990 to 1998, 14 TKR in 7 patients with severe haemophilia were performed. The mean age at operation was 47.3 years (range, 27-62 years). The mean follow-up period was 3.7 years (range, 1-7 years). The TKR was performed bilaterally at the same time in 6 cases. In 1 case, the bilateral TKR was done one after another with 6 months interval. In 10 knee joints, an unconstrained or semi-constrained surface replacement system was used. In 4 joints, a hinged prosthesis was required. RESULTS: The range of motion (extension-flexion) was improved from 0-23-69 degrees preoperatively to 0-4-88 degrees at follow-up. The mean HSS Score increased from 34.5 points preoperatively to 77.9 points at follow up. Perioperatively, no haemorrhages or early infections were observed. In 1 case, 6 years postoperatively a late infection of the hinged knee prosthesis occurred. A two-stage-exchange of the prosthesis was performed. Aseptic loosenings of prosthetic components were not observed. CONCLUSION: The TKR in haemophilia is technically demanding and requires a consequent perioperative F VIII or F-IX substitution. A bilateral simultaneous implantation is useful. The indication for TKR has to be strict because of the higher risks and requires a close cooperation with the haemostaseologist. PMID- 11386101 TI - [Value of ultrasound after endoprosthesis implantation of the knee joint]. AB - PURPOSE: Ultrasound is often applied in the course of treatment after knee arthroplasty, although sonographic normal findings have not been described so far. Characterising these and comparing them to clinical disorders was the purpose of this study. Intra- and extra-articular hematoma and the imaging of the traumatised extension apparatus of the knee joint were of special interest. METHODS: During 4 weeks all consecutive patients after knee arthroplasty were examined clinically and sonographically. Furthermore, the blood parameters were controlled for signs of inflammation or coagulation disorders. These findings were then re-checked before the end of hospital rehabilitation treatment. RESULTS: In all of the patients, ultrasound revealed intra-articular fluid at the beginning and at the end of hospital treatment. The fluid area, differing in extension, was markedly reduced or showed less echogenicity concomitant with an organisation. The patellar tendon in all of the patients showed a loss of echogenicity at the site of operative incision, mostly at the patellar insertion, and a thickening extending throughout the middle part of the tendon even at control. Dynamic ultrasound examination displayed one case of a major defect which had to be revised. Blood parameters of inflammation were decreased, coagulation parameters were normal. CONCLUSION: After knee arthoplasty, extended intra-articular knee hematomas are not rare, only being resorbed or organised to a small degree during a rehabilitation period of three weeks, therefore not yielding relevant information for the course of treatment. CLINICAL RELEVANCE: The patellar ligaments show alterations comparable to acute tendopathy, thus not recommending use of maximal forces or too high stretching of the tendon tissue. Soft tissue defects with the need for interruption of the rehabilitation programme may be detected sonographically. Further studies will be necessary to explore the course of restructuring. Disorders of patellar sliding movement and signs of prosthetic loosening cannot be sufficiently judged at the present time. PMID- 11386102 TI - [The Pediatric Musculoskeletal Functional Health Questionnaire. A function assessment questionnaire for detection of illnesses of the locomotor system in children and adolescents--initial results of validating a German version]. AB - AIM: It was the purpose of this investigation to create a German version of the Pediatric Musculoskeletal Functional Health Questionnaire and to test its reliability, practicability and acceptance in children and adolescents with musculoskeletal disorders. METHODS: In a first step, the Pediatric Musculoskeletal Functional Health Questionnaire was translated into the German language. Then 147 patients with musculoskeletal disorders or respectively, their parents were asked to fill in the questionnaire, in order to test the reliability, the internal consistency, the practicability, and the acceptance of this instrument. RESULTS: The test-retest reliability for the subscales was high, it ranged between r = 0.56 and 0.93. Concerning the internal consistency of items and subscales we found only moderate results. The acceptance of the Questionnaire was high in patients or, respectively, their parents with 92%. Furthermore, the instrument has shown to be practicable and economical. CONCLUSION: With the German version of the Pediatric Musculoskeletal Functional Health Questionnaire a multidimensional instrument is now available that reliably measures health status as well as therapeutic effects in children and adolescents with musculoskeletal disorders. PMID- 11386103 TI - [The Graf standard plane--a "standard sector"? Ultrasound studies of anatomic preparations of hip acetabula of infants]. AB - AIM: To answer the question as to which ultrasound sections, related to the hip joint plane, lead to pictures of the so-called "standard-plan" of Graf, anatomic preparations of infant hips were examined by ultrasound. METHOD: 10 anatomic preparations of infant hips were fixed in a support which allows an exact positioning of the ultrasound plane related to the acetabular inlet plane in a water bath. The joints were examined while changing the relative position in steps of 10 degrees. The ultrasound images were documented by video-print and the alpha-angles were measured if the criterias of the standard plane were fulfilled. RESULTS: In order to obtain images that are analysable by the method of Graf, the ultrasound beam has to intersect with the acetabular inlet plane at defined angles. The acetabular notch has to be anteriorly rotated from the ultrasound beam plane by at least 20 degrees. Beam entry within a 50-degree sector posterior to the perpendicular on the inlet plane gave analysable images. The alpha-angles were markedly affected by coronal-plane transducer tilt. Caudad tilts were associate with lower values, a fact that should be borne in mind in clinical ultrasound investigations. CONCLUSION: The "standard plane" of graf is a "standard sector", which can be defined in all three dimensions. The relations between the acetabular inlet plane and the sonographic beam are described exactly for the first time. The results are important because of their influence on clinical ultrasound examinations. PMID- 11386104 TI - [Incidence of thrombophilic factor V Leiden and prothrombin G20210A mutation in Perthes disease--a pilot study]. AB - AIM: Thrombophilic mutations may play a role in the pathogenesis of the juvenile osteonecrosis of the femoral head, Perthes' disease. We investigated whether children with Perthes' disease have an increased incidence of mutations of factor V (Leiden) and prothrombin (G2O210A), which predispose to thrombosis. METHODS: For this pilot study, we analysed the data of twenty consecutive children (16 boys, 4 girls, mean age at diagnosis 6.4 years). According to Catterall's classification of severity, 2 children were in group 1, 7 in group 2, 8 in group 3, and 3 were in the most severe group 4. Mutations of factor V and prothrombin were identified in EDTA-blood by PCR amplification, digestion with restriction enzymes, and gel electrophoresis. RESULTS: Heterozygoty for the factor V mutation was detected in two children, for the prothrombin mutation in one child. Both results did not differ significantly from the incidence in Germany, which is 0.05 for factor V mutations and 0.04 for prothrombin mutations. CONCLUSIONS: For the presented group of children with Perthes' disease, we did not find an increased rate of factor V or prothrombin mutations compared to the natural incidence. In accordance to other recent studies, our results do not support a link between inherited thrombophilic mutations and Perthes' disease. PMID- 11386105 TI - [Function of back extensor muscles in upright standing position and during sitting with identical back posture--an electromyography study]. AB - PROBLEM: There is a controversial discussion about the loading of the spine by the force of the extending muscles of the back in upright sitting with the back curved as in upright standing. Experimental studies investigating this issue are still lacking. METHODS: In the study presented in this paper we measured in 30 subjects the activity of the extending back muscles both in a natural upright standing posture and in sitting with identical posture of the back as recommended in some back training programs. To this end we recorded the electromyographical surface activity of the back muscles at four levels of the spine in both postures. The EMG activities were intraindividually normalized with reference to the respective activity recorded at maximum voluntary isometric contraction (MVC). RESULTS: At three of the four levels of the spine examined, the group averages of the EMG activity in sitting were significantly higher than in standing (p < 0.001). The maximum activity enhancement evaluated was 48% at Th12. The force of the lower back muscles in sitting with a posture of the back as in natural upright standing could be estimated to be at least greater than 30% of the force at MVC. CONCLUSION: The force needed to enforce in sitting a posture of the back identical to the posture at natural standing entails lower back muscle fatigue in a few minutes. The results of our study agree with recent experimental findings about an increased loading of the spine in lordotic sitting. PMID- 11386106 TI - [Staged reposition and fusion with external fixator in spondyloptosis]. AB - PROBLEM: Is the staged reduction by an external fixateur and combined fusion of slipping vertebra an adequate surgical treatment for lumbar spondyloptosis? METHODS: 11 patients with symptomatic lumbar spondyloptosis were treated using a technique of slow reduction and combined posterior and anterior approach. The first stage consists of a posterior approach with the application of the external fixateur. After staged reduction the anterior and posterior fusion are performed. All patients were pre- and postoperatively classified by radiological and clinical criteria. We report improvements in pain, activities of daily live, and cosmetic appearance. The average follow-up was over three years. RESULTS: Postoperatively one patient failed to reduce, and one developed a subsequent L4/L5 spondylolisthesis. All patients showed a solid spondylodesis with no significant loss of reduction. There was no patient with any neurological deficit. The mean correction of the slipping at follow-up was 84.5%. 9 patients were satisfied with the result. CONCLUSION: With gradual instrumented reduction by external fixateur, an anatomic reduction is nearly obtained and excellent clinical results in the treatment of spondyloptosis can be achieved. PMID- 11386107 TI - [The role of MRI in therapy and follow-up after surgical treatment of osteochondrosis dissecans of the talus]. AB - AIM: Our study was designed to evaluate the use of magnetic resonance imaging (MRI) in the follow-up of surgically treated osteochondrosis dissecans (OCD) of the talus. METHOD: We investigated 16 patients (18 joints) with OCD of the talus, surgically treated in our department between 1990 and 1997. All of them had preoperative MRI scans of the affected ankle. The mean follow-up was 40 months (8 82). All patients were evaluated by clinical examination, plain radiography and MRI using a standard protocol. RESULTS: The clinical Bray score improved significantly from 58 preoperative to 82 postoperative. Using a visual analogue scale we saw a significant reduction of the patients pain level post-op. In 72% of the patients the preoperative MRI was able to predict the accurate stage of the cartilage. Postoperative MRI showed no more lesion in 3, intact articular cartilage in 11, and disrupted cartilage in 4 joints. There was no correlation between clinical, plain radiographical, and MRI findings postoperative. CONCLUSION: Arthroscopy remains the golden standard in evaluating articular cartilage. Using our data, MRI is not the method of choice in the follow-up of surgically treated OCD lesions of the talus. Postoperative use of intravenous contrast media gave no additional information. Postoperative MRI should be reserved only for symptomatic patients to gather additional information about the actual state of the OCD. Afterwards a prompt arthroscopy of the symptomatic ankle should be performed. PMID- 11386108 TI - [Reproducibility of radiological classification criteria of femur head necrosis]. AB - AIM: To assess the reliability of commonly used radiographic features in osteonecrosis of the femoral head. METHOD: For staging of avascular necrosis of the femoral head two different classification systems, Ficat and ARCO, were applied. To measure the size of the necrotic lesions the combined necrotic angle of Kerboul was used. Radiographs of 49 patients with osteonecrosis of the femoral head were graded by two observers at two different time points. The Spearman's correlation coefficient was calculated for intra- and inter-observer reliability. RESULTS: We found good intra- (Spearman's correlation coefficient 0.81-0.87) and inter-observer reliability (0.71-0.74) for the Kerboul's angle. Reliability for the ARCO stage classification was acceptable for intra-observer but unacceptable for inter-observer agreement. The reproducibility of the Ficat staging system was unacceptable. CONCLUSION: The Kerboul angle is a reliable measurement to assess the severity of the osteonecrotic defects in femoral heads. The staging systems of Ficat and ARCO are not sufficiently reliable to assess the disease status of the hip alone. PMID- 11386109 TI - [Differential diagnosis of osteoarthropathia hypertrophicans]. AB - PURPOSE: Chief clinical symptoms of HOA are bone and joint plain, nearly always coupled with finger and toe clubbing. The classification of HOA distinguishes between a very seldom primary (PHOA) and the more frequent secondary hypertrophic osteoarthropathy (SHOA). The pathogenesis of the disease is as yet not wholly understood, but there is a tendency that it may be a bony manifestation of a paraneoplastic syndrome. RESULTS: By means of two cases, one a 72-year old patient with pulmonary carcinoma, the second a 48-year-old patient with a long standing history of COPD, we demonstrate and discuss typical clinical, laboratory, radiographic, and scintigraphic findings associated with SHOA. Primarily the patients consulted an orthopaedic surgeon due to persisting, localised osseous pain. Clubbing of fingers and toes as well as soft tissue swelling usually remain asymptomatic. Characteristically a symmetrical diaphyseal accumulation of periosteal calcification in the vicinity of short and long cortical bones becomes radiographically apparent. Early diagnosis is best facilitated by means of a bone scan, where the symmetrical increase in bone activity and the typical "double stripe sign" can be observed. CONCLUSION: In patients in whom unexplained periosteal reaction is found and phalangeal clubbing becomes evident, the diagnosis of SHOA should be contemplated and further tests performed, so as to rule out a paraneoplastic syndrome. PMID- 11386110 TI - [Scapula alata as a complication of infection with human parvovirus B19 (erythema infectiosum)]. AB - This article reports a case of a 38-year-old woman suffering from erythema infectiosum (slap-cheek disease) complicated by arthritis and severe brachial plexus neuritis resulting in scapula winging. PMID- 11386111 TI - [The "new market" and computer-assisted orthopedic surgery--will CAOS end in chaos?]. PMID- 11386112 TI - [Better than their reputation?--5 to 20 years outcome with single compartment knee joint endoprostheses in medial osteoarthritis of the knee]]. AB - AIM OF THE STUDY: Long-term results of unicompartmental arthroplasty (UCA) in medial osteoarthritis are analyzed and the influencing covariates were explored. METHODS: 261 UCA in 229 patients were evaluated mainly by use of a structured questionnaire, 98.5% of the UCA were analyzed after 10 (5-20) years. RESULTS: In the Knee Sec. Score, the items pain and function were improved significantly. Patients were very content with the UCA. Failures (total 30) were especially caused by destruction of the non-replaced compartment (n = 11) and loosening of the tibial component (n = 5). Survival with aseptic failures as the endpoint was 84.9% after 10 and 74.4% after 20 years. Gender, age at operation, design (St. Georg, Tonnis, Endo, Wessinghage) and height of tibial component could not be proved as determining covariates with sufficient statistical significance. CONCLUSION: Cemented unicompartmental knee arthroplasties show good long-term results in medial osteoarthritis, when the criteria for the indication are strictly respected. PMID- 11386113 TI - [Technical orthopedic shoe management of the prosthetic leg and the salvaged extremity in patients with diabetic foot]. PMID- 11386114 TI - [Diabetic foot. A causality problem in private and mandatory accident insurance]. PMID- 11386116 TI - [Clinical significance of congenital cervical blocked vertebrae]]. PMID- 11386115 TI - [What is reliable in drug therapy of osteoporosis?]. PMID- 11386118 TI - [Radium chloride in ankylosing spondylitis]. PMID- 11386117 TI - [Detection of cortical reorganization after arm amputation and relationship to phantom pain--an fMRI study]. PMID- 11386119 TI - [The impact of developmental psychological and developmental psychopathological aspects on psychotherapy research with children]. AB - The development of efficacious psychosocial interventions for children and psychotherapy for children and adults differ significantly from each other. Nevertheless, interventions have often been developed for adults and--assuming efficacy--generalized for children; as a rule, efficacy studies of interventions with children are implemented like studies with adults. Several studies put evidence on the efficacy of interventions from different therapeutic traditions, but effectiveness studies of psychotherapy--carried out under authentic conditions--revealed less beneficial effects. Important differences between psychotherapy with children and adults are discussed and several aspects are named, which could be responsible for the differing results of efficacy and effectiveness studies. Hence, several implications for psychotherapy studies with children can be derived (particularly concerning developmental psychological and developmental psychopathological aspects). Following issues are discussed: a) influence of maturation and development; b) influencing factors, e.g. characteristics of the clients; c) developmental appropriate methods; d) multimodal and multimethodological interventions; e) utilization of manuals; f) follow-up-studies; g) outcome criteria; h) sources of information. PMID- 11386120 TI - [Sleep deprivation as a predictor of response to light therapy in major depression]. AB - Light therapy (LT) is regarded as the treatment of choice for seasonal affective disorder (SAD). In nonseasonal depression the results of light therapy are nonconclusive. Sleep deprivation (SD), however, is effective in 50-60% of the patients with major depression. The predictive value of Total Sleep Deprivation (TSD) for the treatment outcome of antidepressiva has been already examined. Purpose of the present study was to test whether light therapy is more beneficial in TSD responders than in TSD nonresponders. 40 inpatients with major depressive disorder completed one night of TSD. Twenty TSD responders and 20 TSD nonresponders were randomly assigned to 14 days of bright light therapy (2500 lux, 7-9 a.m.) or 14 days of dim light therapy (red light 50 lux, 7-9 a.m.). Manova with repeated measurements revealed a significant difference in the course of depression over the time between TSD responders and nonresponders, but no significant difference between bright and dim light. Questions of placebo effect, of SAD and of personality variables as predictors of response to SD and LT are being discussed. PMID- 11386121 TI - [Dementia and suicide]. AB - The relationship between suicide and dementia has not systematically been investigated, although the prevalence of both, dementia and suicide, increases with age. In contrast to patients with other psychiatric disorders, patients with dementia were not found to die from suicide more often than expected (SMR, 0). Thus the diagnosis of dementia does not contribute to the elevated suicide risk in old age. In studies using the psychological autopsy method, dementia was rarely diagnosed in suicide victims. Suicide attempts were observed in less than 1% of all patients with dementia. Depression as an important common risk factor of suicide and dementia is often found in patients with Alzheimer's disease (0% to 86%) as well as in patients with multi-infarct dementia (20% to 45%). However major depression was found significantly more often in vascular dementia than in dementia of Alzheimer type. Suicidal thoughts and intents, wishes to die and feelings that life is not worth living were reported in 1% to 42% of all patients with dementia, especially if these patients also suffered from depression. This review comprehensively presents the association between cognitive deficits, insight in early stages of dementia and suicidality and possible confounders which have not systematically been investigated up to now. PMID- 11386122 TI - [Psychiatric emergencies in the physician-based system of a German city]. AB - OBJECTIVE: Pre-clinical Psychiatric Emergency Situations (PES) gain more scientific interest. First investigations revealed them to be the third major cause for emergency physician (EP) calls. However, there is still very little data concerning prevalence, diagnosis and therapy. METHODS: A retrospective analysis of all anonymised EP protocols of the year 1995 in the city of Hamburg was performed. Data of each multiple-choice category was evaluated as well as handwritten and personal notes to determine prevalence, diagnoses and therapy of PES. RESULTS: 2550 out of 26,347 protocols (9.7%) revealed to have a psychiatric disturbance as a major cause for the call. The most frequent diagnoses in the overall young (average age: 43.1 +/- 17.6 years) and male (60.3%) population were found to be alcoholic intoxication (34%), state of agitation (23%) and suicide attempts (22%). In 55% the emergencies could be considered as purely psychiatric; in 35% as a combined somatic-psychiatric emergency. A specific psychopharmacological treatment was not performed. CONCLUSIONS: Psychiatric patients are a considerable group in pre-clinical emergency medicine, however, disturbances are much too rarely documented, diagnosed and treated. Training programs are necessary. Psychiatry has to play a more active role in planning and performing these programs. PMID- 11386123 TI - [Syndromes in environmental medicine: variants of somatoform disorders]. AB - Concerning the syndromes in environmental medicine, like Multiple Chemical Sensitivities (MCS), Idiopathic Environmental Intolerances (IEI), Sick Building Syndrome (SBS), Chronic Fatigue Syndrome (CFS), Candida Syndrome (CS), and Burnout Syndrome (BS), scientific knowledge in etiology, pathology, pathophysiology, diagnosis, therapy, prevention and prognosis is still lacking until now. A critical comparison shows that it is still impossible to find a scientifically satisfying delimitation. Syndromes in environmental medicine show clinical similarities to somatoform disorders. Furthermore, there are the following possible explanations for the existence of these syndromes: Firstly, they may be a complex interaction of environmental impacts, individual predispositions, psychological influences, as well as processes of mental perception and interpretation. Secondly, they may be an effect of distress influenced by culture and social structures and/or thirdly, they may be an latrogenic determination. A more comprehensive characterisation which better considers the complex clinical manifestations is overdue. Although there are neither scientifically validated procedures for diagnosis or therapy nor prophylactic measures, a hardly comprehensible number of partly unvalidated methods is in practical use. Until the syndromes are not finally defined the terms for the syndromes should not be applied to a certain disease. Despite all uncertainities in the evaluation of syndromes in environmental medicine, physicians have the duty to take the affected persons' problems seriously. PMID- 11386124 TI - [Fibromyalgia]. AB - The classification of fibromyalgia is based on the criteria of the American College of Rheumatology. For diagnostic reasons autonomic disturbances and mental features have to be considered. The distinction between fibromyalgia (tender points) and myofascial pain syndrome (trigger points) is essential. Internal and neurological disorders as a primary cause of fibromyalgia have to be excluded. The aetiology and pathogenesis of fibromyalgia still remain uncertain. The myopathological patterns in fibromyalgia are non-specific: type-II-fiber-atrophy, a slight increase in lipid droplets, a proliferation of mitochondria and a slightly elevated incidence of ragged red fibers. Biochemically alterations of the serotonin system and high levels of substance P in the cerebrospinal fluid of fibromyalgia patients are important. Animal experiments showed that the central stimulation by nociceptor input from muscles is exaggerated in skeletal muscle pain conditions, suggesting central hyperexcitability. The diagnosis of fibromyalgia requires a thorough exclusion of other rheumatologic and neurologic disorders. The differential diagnosis is complicated by an overlap to other chronic somatoform pain disorders. PMID- 11386125 TI - [Diagnosis of normal and abnormal delayed hypersensitivity to Candida albicans. Importance of evaluating lymphocyte activation by flow cytometry]. AB - Delayed type hypersensitivity (DTH) to Candida albicans is commonly observed in human. Abnormal DTH has already been described but its diagnosis is difficult to ascertain. We present now a clinical and biological study in 60 patients with a clear distinction between these two kind of Candida albicans DTH. Clinical abnormal Candida albicans DTH was characterized by a syndromic reaction 24 to 48 hours after intradermal injection. This reaction was characterized by an exacerbation of clinical symptoms. In vitro, activation of whole blood with Candida albicans antigen was detected by using flow cytometry after staining for activating markers. CD 25 positive T cells were detected in a 7 days culture in all patients. Percentage of CD 25 positive T cells was correlated to the intensity of the local cutaneous DTH reaction. CD 69 positive T cells were detected after a one day culture only in patient who presented a syndromic reaction to intradermal injection. PMID- 11386126 TI - [Acetylsalicylic acid: hypersensitivity, intolerance, or allergy?]. AB - After a century of therapeutic use of acetyl salicylic acid (ASA), the different mechanisms involved leading to the therapeutic activity and to the various clinical reactions (even life threatening) observed in patients sensitive to ASA or in general to the non steroidal anti-inflammatory drugs (NSAID's) are still to be discovered or confirmed. During the last century a lot of scientific results have been published in favour of a wide panel of mechanism including an humoral (IgE and/or IgG) or a cellular (lymphocytes) immune reaction, inhibition of the cyclooxygenases, a cellular activation (mast cells, basophils, eosinophils and platelets), a competition at the serum albumin level and an activation of the complement cascade. In general the same actors than in the anaphylactic reaction are found (mediators, cellular activation and amplification systems). It is clear today that these reactions for which an accurate diagnosis is mandatory cannot be explained by a single mechanism, which is the case of a lot of other allergens. PMID- 11386127 TI - [Anti-allergy therapy in children]. AB - The discovery of the physiopathological mechanisms of asthma and the diseases of allergy has allowed the development of many symptomatic therapeutical perspectives. We review below the mechanisms of actions, routes of administration, secondary effects, dosages and indications for different commercialised anti-allergy medications. A better understanding of the pharmacology of the different medications should produce optimisation of the treatment of the allergic child. PMID- 11386128 TI - [Individualized Reception Project at school and for school food services for children with a food allergy]. AB - The circulars of 1992, 1993 and more recently of 10th November 1999, illustrate the significant progress of schools in the reception of sick children and the help in execution of the medical regulations. The National Education physician is a privileged interlocutor of families, the treating physician and the education authority to make the link between the caregivers and the school. Application of the 1999 circular, which has the objective of taking into account the food allergies in the 1993 plan of action, raises difficulties of three types: Difficulties of a medical type raised by the frequent confusion between the Protocol for urgent intervention and the individualised reception plan. The Protocol for urgent intervention, of which the National Education physician is the recipient, must be accompanied by a letter from the allergy physician that takes into consideration simple immunological sensitization and true allergy and to appreciate the risks that concern food disparities that are not always the same. The almost systematic prescription of adrenaline in protocols for urgent intervention, without argument and in the absence of commercialisation of an auto injectable pen at the time of distribution of the circular, was a source of resistance by education authorities to catering services. Difficulties of a human type concerning the creation of fear in families, in the education authorities, in the catering services and the time-lag between some families and their careful vigilance with regard to the dietary observance and the conduct to be taken in the case of acute symptoms. Difficulties of administrative or legal types knowing that the scholars catering is a service which does not have the obligatory character of the teaching plan, and which accepts the sole competence of the authorities for 1st degree education. In total the teaching plan and pupil catering for children with a food allergy needs making the parents aware of the responsibility between the allergy physician and the National Education physician who have equal responsibility. With clear and well argued diagnostic and therapeutic information the National Education physician will be able to contribute to the installation of a calm climate that is indispensable to a quality reception. PMID- 11386129 TI - Food anaphylaxis induced by aspirin. PMID- 11386130 TI - Keeping faith in the community. Not-for-profit hospital companies find way to sell without selling out. PMID- 11386131 TI - Watching out for nursing home residents. Cameras could help curb abuse but others argue they invade patient privacy. PMID- 11386132 TI - Triad lightens its load. Two more Quorum hospitals are up for sale; quarter net income down a bit. PMID- 11386133 TI - Playing monopoly? FTC probes mammoth drug distributor merger. PMID- 11386134 TI - Woe is us. Stinging reports criticize health industry, detail widespread dissatisfaction. PMID- 11386135 TI - The science of giving. PMID- 11386139 TI - Coalitions lose mission. New studies show purchasing alliances fail to secure lower healthcare premiums. PMID- 11386140 TI - An uncomfortable cap. Obscure law limiting debt a 'constant irritation' for systems trying to merge. PMID- 11386142 TI - [Illness theory and identity of psychiatry]. PMID- 11386143 TI - [Structural brain changes in patients with schizophrenic psychoses. From focal pathology to network disorder]. AB - Schizophrenia is a brain disorder characterized by a heterogeneous clinical symptomatology. Accordingly, many structural brain changes are not associated directly with clinical symptoms. These structural changes can be detected in the frontotemporal cortex and may correlate with the course of the disease. The most important etiological concept is the neurodevelopmental hypothesis according to which developmental, morphologically detectable changes predispose for the acquisition of schizophrenia. The relevance of neurodegenerative components also remains to be determined. However, it is becoming increasingly clear that schizophrenia is not associated with pathological changes in a circumscribed brain region but with widely distributed morphological changes. Presently, the leading hypothesis for explaining these changes is a frontotemporolimbic network disturbance with cytoarchitectural changes in the heteromodal association cortex. Present research therefore focuses on testing this theory using functional imaging on a macroscopic level and examination of the neuronal cytoarchitecture on a microscopic level. PMID- 11386144 TI - [Disordered behavioral control in psychiatric illnesses. Neurophysiological aspects of impulsive behavior]. AB - OBJECTIVE: Dysfunction of behavioral control represents a central component of many psychiatric disorders and may be connected with self-mutilating and antisocial behavior. However, only a few studies have investigated objective assessment and quantification of impulsive behavior, which is relevant for psychiatric practice and research. Because new techniques enable objective assessment of behavior generation and execution, the current knowledge about neurophysiological aspects of behavioral control is summarized. METHOD: The contemporary literature on neuronal mechanisms of initiation and execution of behavior and movements and new neurophysiological methods and findings are discussed in respect to their application in psychiatry. RESULTS: Behavior, defined as movements of a body in time and space, is the result of cortical and subcortical processes. These consist of two general components: (1) perception and selection of a stimulus and (2) execution of a movement, including decision and planning phases. Planning and execution, modulated by cognition and emotions, are dependent on the balance of inhibitory and excitatory aspects of behavior control. Combined detection of electrical brain activity and kinematic analysis of peripheral movements enables the analysis of control mechanisms. CONCLUSION: Impulsiveness and inhibition of behavior depend on neuronal control mechanisms which can be analyzed by neurophysiological methods. Objective assessment of impulsive behavior related to psychiatric disturbances including eating disorders, substance abuse, and personality disorders may clarify underlying neurobiological mechanisms and enable rational pharmaco- and psychotherapeutic interventions. PMID- 11386145 TI - [Schizophrenia and delusions in middle aged and elderly patients. Epidemiology and etiological hypothesis]. AB - Knowledge of the similarities and differences between early- and late-onset schizophrenia and between late-onset schizophrenia and paranoid disorder of old age and very old age is fragmentary. We compared diagnosis, subtypes, syndromes and symptoms between first-episode schizophrenia (ICD-9: 295) and paranoid disorder (ICD-9: 297, 298.3/4.) over the life cycle in a population-based (N = 232) and a clinical first-admission sample (N = 1109). Apart from different age patterns of the sexes only two symptom groups were significantly different between early- and late-onset illness: paranoid and systematic delusions showed a linear increase, symptoms of disorganisation a linear decrease over the life cycle. Clearly different between early- and late-onset illness were the neurobiological and psychological risk factors, suggesting that both neurodevelopmental and neurodegenerative disorder causes psychopathology typical of schizophrenia. Late- (40 to 60) and very-late-onset (over 60) cases of both groups of illness showed the same symptom profiles, merely the number of symptoms being higher in the group diagnosed with schizophrenia. Age was the only factor significantly contributing to a clinico-diagnostic differentiation of schizophrenia from paranoid disorder beyond age 40. PMID- 11386146 TI - [Uniformity and diversity of the human image in scientifically oriented human medicine]. AB - Nowadays, hardly anyone would oppose the demand for more rationally based medicine--the catchword here being "evidence-based medicine" (EBM). Anyone trying to comply with this demand will be faced with the question of what is meant by "evidence". It would be false to think that rationality is guaranteed by applying the Galilean method of exact induction. Exact induction aims at objective propositions free of subjectivity. Such propositions are regarded as generally valid, or "true". They lead us toward the transcendent platonic "idea", which is by definition beyond our reach. Exact induction enables us to derive representations of a transcendent idea by means of experimental research. These representations may or may not be useful in actuality, for example in medical therapeutic strategies. Strictly speaking, reproducibility, which is generally taken as proof of a given hypothesis, is not equivalent to identity. Identity implies, among other things, simultaneity. PMID- 11386147 TI - [Quantitative detection of dyssocial and psychopathic personalities in criminal expert assessment. Retrospective study of the value of the German version of the Hare Psychopathy Checklist]. AB - In 1980, the Hare Psychopathy Checklist (PCL), an operationalised rating scale for the quantitative assessment of "psychopathy" as seen in the Anglo-American literature, was developed by R.D. Hare in Canada. It proved to be the most valid prognostic instrument for assessing relapse of criminals in English-speaking countries. This study was aimed at verifying the applicability of a German version of the revised PCL (PCL-R) on carefully selected subjects from the Forensic Psychiatric Department of the Psychiatric University Hospital in Munich. Two groups with an equal number of different forensic-psychiatric reports were evaluated retrospectively using the PCL-R. The results confirm the applicability of the PCL-R for the assessment of psychopathy in our sample population. One can expect that the positive experiences with the PCL-R abroad can be transferred to German conditions. The results are encouraging as to the practicability of the PCL-R as a prognostic instrument in routine forensic assessment. PMID- 11386148 TI - [Pica as a harbinger of schizophrenia]. AB - The author reports the case of a youth who first became salient by self-neglect and finally was hospitalized for heavy drug abuse. In the closed ward, he suddenly displayed pica behavior but, in the further course, developed a typical schizophrenic syndrome. The diagnostic criteria are presented as well as all subtypes of pica. The author goes into the question of the aetiology of this enigmatic disorder. Finally, he points out the typical complications of pica and the therapeutical possibilities which are determined most of all by the underlying disease. PMID- 11386149 TI - [Heautoscopy. Delusions of a double and rare hallucinations of personal gestalt]. AB - In the psychiatric literature, the phenomenon of the doppelganger or double is characterized into different types of delusional misidentification syndromes and heautoscopic symptoms. Specifically, we focus attention on heautoscopy, which might also be named "dissociative" after excluding an organic etiology. We report a case of heautoscopy in a woman and discuss the clinical variability of this psychopathological phenomenon. Furthermore, a short treatment of the historical and literary implications of this delusion is presented. PMID- 11386150 TI - [Successful treatment of simple schizophrenia with atypical neuroleptics (clozapine, risperidol)]. AB - Schizophrenia simplex is a negative subtype of schizophrenia with a low prevalence but poor prognosis. We report on a 22-year-old male who responded well to atypical neuroleptics (clozapine and risperidone). PMID- 11386151 TI - [Diffusion tensor imaging (DTI) and functional magnetic resonance tomography (fMRI) expand methodological spectrum in psychiatric research]. AB - In psychiatric research, there is a growing interest in the microstructural and functional characteristics of brain networks, which often form the basis of current etiological concepts. As a result of novel magnetic resonance imaging techniques, the pathogenic characteristics of neuronal activity and connectivity can be examined in a noninvasive, safe, and repeatable manner. Functional magnetic resonance imaging (fMRI) uses blood oxygenation level-dependent (BOLD) measures for identifying the gray matter contribution to cognition. Diffusion tensor imaging (DTI) reveals the course and structural integrity of white matter projections. Because DTI does not require special motivation and performance, group differences in psychiatry are more easily interpreted in terms of underlying pathology. To date few studies have tried to investigate both, i.e. dynamic and microstructural data in the sense of a modern multi-dimensional investigation approach. The combination of both techniques, however, seems to offer a promising vehicle to further extent our current understanding of mental disorders and to identify populations at risk. In addition to addressing findings in psychiatric research, the present article presents a technical overview of DTI and examines the limitations and potential applications of both techniques. PMID- 11386152 TI - [Public press commitment of the DGPPN (German Society of Psychiatry, Psychotherapy and Neurology]. PMID- 11386153 TI - [Sleep stages after psychoemotional exposure: individuality-dependent changes]. AB - Sleep stages were studied in healthy subjects with the aid of a battery of tests involving questionnaires, psycho-tests, motor tests, polysomnography, and cardiomonitoring. An induced psychoemotional tension was shown to change the 1st sleep stage, to decrease percentage of the 2nd stage of the slow-wave sleep, to redistribute the delta-sleep, and to suppress the REM sleep mechanism. The cerebro-visceral function of increasing the heart rate and its variability in night sleep was also affected. Patterns of the sleep structure changes depended on personality characteristics of the subjects. Thereupon, individual programs should be used in studies of psychoemotional stress effects upon the sleep pattern. PMID- 11386154 TI - [Biphasic response of the human nervous system to geomagnetic storms studied by EEG]. AB - In two subjects: a male phlegmatic, 56, and a female melancholic, 22, the EEG was recorded at resting. The EEG patterns were juxtaposed with the geomagnetic activity index Ar and the solar activity index SF, as well as between themselves. The results revealed qualitatively similar biphasic responses: a generalised diminishing of the EEG spatial synchronisation and, on the next day, a generalised augmentation of the phenomenon, as compared with a prolonged quiet period. A general unspecific stress response is supposed to underlie the aforementioned dynamics, whereas a reduced cortical tone during a magnetic storm and an enhanced one after its cessation correspond to the two phases observed. Specifics of responses in both subjects corresponded to their individual profiles of interhemisphere asymmetry. PMID- 11386155 TI - [Swimming stress-induced histochemical and morphological changes in various regions of the rat hippocampus]. AB - In rats, swimming stress enhanced functional activity of the hippocampus' pyramidal neurones as manifested by diminishing of the glycogen contents, increasing amount of nucleic acids, augmented nucleus/cytoplasm ratio. PMID- 11386156 TI - [Feedback in the stimulus-response system determines stress specifics]. AB - The paper surveys controllability in the stress theory. Uncontrollable action is perceived by living organisms in a principally different way than the controllable one and induces persisting pathological changes. The controllability, i.e. presence of a feedback in the stimulus/response system, is determined by an interaction of the medium factors and a strategy of subject's behaviour. A passive strategy proves to be optimal for adaptation under the conditions when an animal cannot change unfavourable medium conditions. The authors base their theoretical statements on their own experimental studies as well as literature references analysis. PMID- 11386157 TI - [Cell activation in the hypothalamus after exposure to an antigen (based on c-fos gene expression)]. AB - Increase of the c-fos mRNA positive cells number was insignificant in 30 min. following activation of the rat hypothalamic structures with the tetanus toxoid (TT). Elevation of the c-fos mRNA positive cells number occurred in the hypothalamic' posterior (PHA), lateral (LHA), anterior (AHA), areas dorsomedial (DMH), and ventromedial (VMH) nuclei within 2 hours of the TT administration. In 6 hours the c-fos mRNA positive cells number decreased in PHA, LHA, DMH. The c fos mRNA expression was stable in arquate and supraoptic hypothalamic nuclei following either the TT or saline administration. PMID- 11386158 TI - [Input impedance for studying hydraulic parameters of the vessel system]. AB - Vascular input impedance can be used as an effective tool in estimating hydraulic parameters of arterial bed. These parameters may be interpreted as hydraulic resistance, elastance and inertance of particular sites of the arterial system. There is no significant difference between these parameters and those obtained through a direct measurement. PMID- 11386159 TI - [Effect of acute congestive splenomegaly on erythropoiesis in rats]. AB - Spleen veins ligature (SVL) led to acute congestive splenomegaly in rats with subsequent normochromic anaemia disappearing on the 21st day after the SVL. An appreciable depression of the bone marrow erythropoiesis, particularly of the so called "proliferating erythroblastic islets" number, was evident on the 7th and 14th days of post-SVL period. The post-SVL anaemia was also associated with occurrence of the "islets" missing central macrophages and eosinophil-enriched "islets" in bone marrow. PMID- 11386160 TI - [Participation of various types of serotonin receptors in the regulation of drinking behavior and salt appetite in rats]. AB - The 5-HT1A agonist 8-OH-DPAT was shown to diminish the water and 1.8% NaCl solution consumption, whereas its antagonist pMPPI--to enhance the water intake in rats. Another agonist: CGS-12066A enhanced water intake or exerted no effect on drinking behaviour and diminished salt intake. Other substances under study exerted various effects. The data obtained suggest that the 5-HT receptors are involved in regulation of water and salt intake, even though the mechanisms of the effects are different. Apparently 5-HT1B and 5-HT2A receptors play an activating role in regulation of water intake, whereas 5-HT1A, 5-HT2C and 5-HT3 receptors act as inhibitory ones. Only three of the receptors under study seem to regulate salt intake by inhibiting the salt appetite. PMID- 11386161 TI - [Involvement of the nucleus accumbens in immune response stimulation following activation of opioid mu-receptors by DAGO]. AB - The data obtained suggest that the agonist of the nucleus accumbens' mu-opioid receptor DAGO significantly increases the number of plaque- and rosette-forming cells following immunisation with sheep red blood cells in sham-operated Wistar rats. Bilateral destruction of the n. accumbens inhibited the immune response. PMID- 11386162 TI - [Effect of cultivation conditions on the functional activity of thyrocytes in the organ culture of the thyroid gland in newborn piglets]. AB - In new-born piglets' thyroid gland culture, thyrocytes were capable of synthesising thyroxine and triiodothyronine for over 12 days. A 48-hour decline of the medium pH to 6.45 did not affect further functional activity of the cells. Decrease in temperature to 26-28 degrees C augmented the thyroid hormones level as opposed to continuous cultivation at 37 degrees C. Degradation of the thyroid gland follicular structure induced a change in the hormones biosynthesis. The thyrocytes not arranged in the follicles mainly produced triiodothyronine in concentrations 5 to 8-fold higher than the T4 contents. PMID- 11386163 TI - [Effect of pyracetam on fatigue of inspiratory muscles in cats]. AB - The aim of the work was to study the influence of pyracetam on respiratory muscles fatigue and ventilatory disorders caused by inspiratory resistive load in cats. The experiments have show that after the use of pyracetam in conditions of fatigue total bioelectric activities of inspiratory muscles and of the phrenic nerve and transdiaphragmal pressure restore; duty cycle, respiratory rate and tidal volume per minute decrease. The conclusion is drawn that pyracetam in the dose 300 mg/kg, in intravenous administration, compensates inspiratory muscle fatigue at the expense of its central mechanism of action. PMID- 11386164 TI - [Response of motor units of the abdominal musculus rectus to afferent input in the course of the long-lasting activity in rats]. AB - In rats, in the course of a long-lasting m. rectus abdominis motor units activity combined with repetitive afferent stimulation of the ischiadic nerve, the units' firing rate decreased. The dependence of the motor units response to afferent stimulation on the background firing rate was preserved during the long-lasting activity in spite of changes in the background firing rate. PMID- 11386165 TI - [Cold-induced paralysis and recovery of lung respiration and heart contractions in newborn rats (inversion of Arrhenius law for physiological functions)]. AB - Arrest of respiration and heart activity in new-born rats aged 3-4 days and 10-11 days was shown to occur at a body temperature 6-7 degrees C and 2-3 degrees C lower than in adult rats, resp. At room temperature the body temperature of profoundly cooled rat's litter gradually increases and the functions are restored. In 3-4-day old rats, at the body temperature rising from profound cooling to 15-18 degrees C, the respiration and heart rates are 2-4-fold more than at the same temperature attained from the normal body temperature. These differences in the respiration and heart rates at the same body temperature suggest an inversion of the Arrhenius law (the Q10 coefficient) for physiological functions in early ontogenesis. This effect completely disappears in 10-11-day old rats. PMID- 11386166 TI - [Effect of histamine on duration of phases of the respiration cycle]. AB - Histamine produced either a bronchodilation or a bronchoconstriction in rats. In a 0.01-1.0 mcg/ml concentration histamine augmented the contractions amplitude in electrical stimulation of the trachea, in a 10-100 mcg/ml concentration histamine enhances the muscle tone thus decreasing the induced contractions. The histamine effects seems to be connected with its prevailing influence on different structures of the airways depending on the concentration. Its high concentrations act directly on the smooth muscle whereas its lower concentration affects receptors signaling the functional modules of the metasympathetic nervous system within the intramural ganglia of the trachea. PMID- 11386167 TI - [2d European congress on biogerontology]. PMID- 11386168 TI - [I. P. Pavlov's library]. PMID- 11386169 TI - [Biomechanical evaluation of osteosynthesis with cannulated screws for Ponteau Colles fractures in a bird model]. AB - INTRODUCTION: The use of cannulated screw without protrusive head in Colles fractures could avoid some of the drawbacks of conventional pinning. In order to compare the mechanical resistance of Kirshner wires versus cannulated screws we designed a animal model of Colles fractures and tested three types of osteosynthesis: A: K-wires, B: Herbert cannulated screw, C: specific cannulated screws. METHOD: After creating a 10 mm defect in three sets of 10 fresh turkey tibia, 3 types of osteosynthesis were done and tested in compression with an Adamel Lhomargy machine: set A: K-wire fixation, set B: Herbert screws fixation and set C: specifics screws fixation. RESULTS: The compression strength needed for failure of the fixations were: for the K-wires (set A): 52 N +/- 17; for the Herbert screws (set B): 93 N +/- 39; for the specifics screws (set C): 160 N +/- 48; (p < 0.0001). DISCUSSION: The use of an animal model makes the experimentation easier and the sampling more homogeneous. In this model, resistance to compression of the cannulated screw was better than K-wires and the specific cannulated screw better than Herbert screw. Therefore clinical trial of osteosynthesis with cannulated screw in Colles fracture could be considered. PMID- 11386170 TI - [Professional absenteeism and surgery for carpal tunnel syndrome. Results of a prospective series of 233 patients]. AB - AIM: The aim of our work was to evaluate the influence of the type of patient's insurance ant the return to work interval after carpal tunnel release. MATERIAL AND METHODS: An prospective analysis of 233 patients (18-65 years old, full time workers) undergoing a carpal tunnel release between January and June 1998 was conducted to determine the interval between surgery and return to work. For statistical analysis we used the Ms Works and Status software package. The mean return-to-work intervals were determined for the following and compared for three groups. Group 1: independent worker, n = 87; group 2: wage earner, n = 90; group 3: civil servant, n = 56; and 4 others subgroups: manual workers, n = 164; non manual workers, n = 69; patients with social security insurance, n = 191; patients with workers compensation, n = 42. RESULTS: For the patients in group 1 the average return to work was 17 days (11 days for non manual workers, 29 days for manual workers). In group 2, the average was 35 days (21 days for non manual workers, 42 for manual workers). In group 3 the average was 56 days (49 days for non manual workers and 63 days for manual workers). The statistical analysis showed the civil servant took significantly longer to return to work than independent workers or wage earner (p < 0.05). The work related patients took significantly longer than patients covered by social security. The effect of occupational han (manual versus non manual) use was clear in the group 1 and 2, but there was no difference in the group 3. DISCUSSION: Our study of 233 patients demonstrated significant difference between independent workers, wage earners, and civil servants in term of return-to-work intervals. In comparing manual and non manual workers, we found a significant difference in group 1 and 2, but in civil servants group non difference were found. We cannot explain these findings on medical grounds. PMID- 11386171 TI - [Knowledge of flap complications in finger pulp surgery and patient education- ethical implications]. AB - INTRODUCTION: The two highest French courts have recently decided that not to fully inform a patient constitutes negligence, and that doctors should be able to prove that the information has been given. The medical community is now debating as to how to protect physicians without harming the doctor-patient relationship. Following this study we put forward the hypothesis that although information might not be available to surgeons nevertheless this should not prevent them taking the ethical steps necessary to redress the situation. METHODS: We conducted a telephone interview to discover what surgeons knew about the complications of finger flaps used for the reconstruction of pulp defects. Four flaps were studied: the VY flap described by Atasoy, the island finger flap known as Ventataswami's, the reverse island flap and the thenar flap. Surgeons were asked to give the percentage of necrosis, infection, sensory disturbances, digital stiffness and exclusion, as well as the time off work, the time of healing and the sensory discrimination of the flap. The results obtained were compared to those obtained by a literature review. RESULTS: Twenty one qualified hand surgeons answered the questionnaire. Many answers were absent or only fragmentary when sought in the literature. Where complication rates were available, there was a huge variation among series. A large variety of answers was also found in our study. The direct island flap was the best known flap. Complications of the reverse island flap and the thenar flap were overestimated in the group interviewed. DISCUSSION: There is no true answer, either in the literature or from the study group. There is a gulf between the jurists' demand and the possibilities of knowledge by surgeons. We are still waiting for clinical studies that will answer the questions asked by patients regarding the complications of surgery. However, even if absolute knowledge is not available, it is the ethical duty of surgeons to give the patient information that is as close as possible as the "truth", and that will be helpful to them. PMID- 11386172 TI - Intercostal nerve transfer classification. AB - We report a new classification of intercostal nerve transfer. The specific application in brachial plexus reconstruction is described. This method was applied in intercostal nerve transfer to the musculocutaneous nerve in 15 patients. In type 1, "MOTOR to MIXED" transfer one harvest only the intercostal main motor branch which is connected at the trunk of the musculocutaneous nerve, without fascicular specificity. In type 2, "MIXED to MIXED" transfer, based on neural cartography, the main motor branch and its sensitive accessory anterior branches are connected to the musculocutaneous nerve trunk, in attempt to connect the motor fascicules together at the center and also the sensory fascicules together, at the circumference. In type 3, "MOTOR to MOTOR" transfer, the intercostal main motor branch is connected to the musculocutaneous motor branch directly destined to the biceps brachialis. The aim of this work is to evaluate the results between different series. PMID- 11386173 TI - [Use of distally based intermetacarpal flaps to cover dorsal traumatic defects of the middle fingers]. AB - INTRODUCTION: The presence of distal intermetacarpal anastomoses between dorsal and palmar vascular networks makes it possible to dissect and isolate Distally Based Dorsal Hand (DBDH) flaps. Quaba and Davidson first described this possibility. The purpose of this paper is to report our experience using the DBDH flap to reconstruct complicated defects of the long fingers. MATERIAL AND METHOD: We are reporting our experience in eight cases (seven performed in emergency) where such a flap was used to cover dorsal traumatic skin defects of the long fingers. The average age was 40.3 years, ranging from 25 to 67 years. All the patients were males, seven of them had a job at the time of injury and the last one had already retired when injured. RESULTS: All the eight flaps survived, sometimes with minor complications. Only one case developed a whole-thickness distal necrosis treated successfully by regular dressings. In one case, a Z plasty was performed to correct a retraction in the 2nd web-space two months after surgery. DISCUSSION: Many flaps have been described to cover dorsal skin defects in the fingers. All of them display some advantages and some disadvantages. The use of this flap allows coverage of vast skin defects and thereby early finger mobilisation. The surgical technique is rather easy, and it does not require microsurgical experience. Skin grafting to cover donor site is not always necessary. PMID- 11386175 TI - Right and left traumatic fractures of a bilateral spontaneous fusion of the elbow. A case report. AB - Report of an exceptional case of spontaneous ankylosis of both elbows, without any family history, trauma, infection or rheumatological disease. An incomplete form of a congenital known syndrome or a thermal reflex sympathetic dystrophy were the most plausible etiologies. The traumatic fractures of both radio ulnohumeral synostoses have been treated conservatively, within a five years interval period. The arthrodesis of both elbows at 35 and 80 degree of flexion did not impinge on the life autonomy and the work ability of the patient. PMID- 11386174 TI - [Long term results of the surgical treatment of bicondylar fractures of the distal humerus extremity in adults]. AB - PURPOSE OF THE STUDY: In intra-articular fractures of the distal humerus, full functional recovery is difficult to obtain. An osteosynthesis by plate is the treatment of choice, but location and type of plate always remain open for debate. We present a consecutive series of intra-articular fractures of the distal humerus treated by osteosynthesis. The aim of the study is to determine and to compare the results of various types of osteosynthesis. MATERIALS AND METHODS: We reviewed 55 patients at an average of 108 months after early internal fixation for intraarticular displaced fractures of the distal humerus type C according to the A.O. classification. Intraarticular osteotomy of olecranon was used in 37 subjects (67.27%). The osteosynthesis has been achieved with a precasted lateral plate for 31 patients, with two posterior plates in 18 subjects, with a screwing on triangulation in four patients and with pins in two patients. RESULTS: The osteosynthesis with two posterior plates obtained a good result between 78.57% and 92.86% of cases, whereas the osteosynthesis with a precasted lateral plate gave a good result between 73.68% and 76.32% of cases according to the score used for estimation. In the aggregate, the functional estimate has a good result in the most of the cases whatever the score of estimate. At follow up we observed an average range of motion of 103 degrees. This value is quite good because it corresponds to the sector of the useful functional mobility. DISCUSSION: The review of our cases and the literature prompt us to follow the way of the osteosynthesis in adjusting indications to the type of fracture and in using a well-codifed technology. As it is already the case, a minimal osteosynthesis by screwing or by pins must be left except for peculiar cases in osteopenic elderly patients and if they have a very poor health. Contrary to other studies, the osteosynthesis with two posterior plates has given us better results provided that it has been systematically associated with a triangulation screwing in order to increase the strength of fitting in the sagittal plan. The precasted lateral plate gives a stable fitting too, nevertheless it is well advised to associate it to an osteosynthesis of medium column especially when the fracture is type C3. We have statistically proved that the age is not contraindication for osteosynthesis. CONCLUSION: This surgery is difficult and entails complications. The dismantling of the synthesis always gives poor results particularly if it is succeeded by an immobilization in plaster. The poor reduction, origin of arthrosis, of loss of bony substance and of calcifications worsen the functional prognosis. Last but not least, a good result can be obtained in the most of the cases however the types of fractures. PMID- 11386176 TI - [Osteogenic exostosis of the capitate bone. Case report and review of the literature]. AB - The occurrence of an osteochondroma in the carpus is very rare and its excision is indicated in the case of significant symptoms or change in its appearance. The diagnosis is often made in adulthood due to the onset of a functional problem even though development of the tumour occurs during skeletal growth. We report the case of a 38 year old patient, with no antecedent trauma, who presents with simultaneous exostoses on the dorsal and palmar surfaces of the capitate, which has not been previously described in the literature. The existence of a bipolar lesion extending anteriorly and posteriorly in the carpus is a possibility which may not be apparent and renders plain radiograph insufficient in the investigation of such a lesion. CT scan and MRI scan are indispensable in the investigation of this kind of carpal lesions, allowing better visualization of the base of the tumour, the expansion of the tumour and relation to the neighbouring soft tissues and the presence of malignant degeneration. PMID- 11386177 TI - [Chondrosarcoma of the hand. Diagnostic and therapeutic discussion (three case reports)]. AB - Chondrosarcoma of the hand is very rare. We report three observations which enable us to make a review of its pathology. There were two men and one woman aged 72, 38 and 63 years respectively. Localisation was phalangeal in two cases and metacarpal in one case. Two cases were central and one peripheral. Radiographs were characterised by the presence of lytic areas with intralesional calcifications and soft tissue extension. Histological diagnosis was made by biopsy in two cases and after amputation of the finger for lesional spread in one case. Treatment consisted of amputation of the finger in two cases. In the case of metacarpal localisation, wide resection of two rays was followed by recurrence after ten months which required amputation of the hand. We agree with other authors who believe that chondrosarcoma in the hand is due to malignant change in a pre-existing chondroma. Conservative treatment is difficult due to the small size of the hand which allows easy spread of the tumour from compartment to compartment. PMID- 11386178 TI - [American Society for Surgery of the Hand. ASSH. Seattle, 5-7 October 2000]. PMID- 11386179 TI - Chemical technology for the toxic gas flow control through process water system. AB - The aim of this work is focused on the safety and toxicological aspects due to under-pressure industrial plant management, above all in the case which the gas is very dangerous for human health and environment. Here is illustrated the safe method of control of risks through specific choices of engineering devices and chemical process: in this way we have shown the mathematical calculation regarding the case of ammonia flow gas running in the piping and plant under pressure. In this paper the Authors show the assessment of the technological solution for falling down of a toxic gas as NH3, which lets off from safety values facilities. The under pressure industrial plants with ammonia are protected through the safety valves, settled at 20 bar pressure. The out-let gas flow is capted by a tank of a water bulk of five time theoretical water amount necessary to the complete absorption of gas. In order to prevent any health risk and carry out a safety management, it needs to verify two basic aspects, with connected specific techniques: 1. The safety valves technology through the mathematical calculation of operating device; 2. The absorption process of the toxic agent for controlling of dangerous runaway of gas. PMID- 11386180 TI - [Low flow anesthesia and environmental pollution]. AB - AIM OF THE STUDY: The aim of the present study is to evaluate operating room pollution and isoflurane and N2O consumption in open system and low flow anesthesia. MATERIALS AND METHODS: Environmental concentration of N2O and isoflurane were measured in an operating room of 150 m3 with 10 air changes by an infrared gas analyser (Bruel & Kjaer) in open system and in low flow anesthesia. Accurate measurement of ihalatory anaesthetic and N2O consumption was made. RESULTS: Concentrations of N2O and isoflurane in operating room were 4.83 ppm and 0.4 ppm respectively, lower compared with open systems: 301 ppm and 11.1 ppm respectively. Isoforane consumption was 25 cc in five hours during low flow anaesthesia, lower compared with open system: 125 ppm, N2O consumption was 4 l/m in open system, 1 l/m in low flow anaesthesia. PMID- 11386181 TI - [A case of pleural mesothelioma caused by unusual occupational exposure to asbestos in the wool industry]. AB - We report the case of a 73-year-old worker who died of pleural mesothelioma, after being employed for 35 years in a wool textile plant of Biella (Italy). Close investigations revealed that he provided for the maintenance of materials and machines. In particular, he used to replace asbestos parts such as rings, joints and insulations of pipelines (dyeing unit), as well as brake linings of warping, looming and combing machines. Beside confirming the importance of an accurate occupational anamnesis to recognize work-induced cancers, the case draws the attention on the risk of mesothelioma in the wool industry, an occupational setting that is not usually considered as a potential source of exposure to asbestos fibres. Such pollution might explain the increased mortality due to pleural mesothelioma in the Biella area (characterized by a prosperous textile industry), reported in previous epidemiological studies. PMID- 11386182 TI - [General criteria of working ability in subjects with color vision defects]. AB - A correct evaluation of the colour vision is important to make decisions concerning the recruitment in numerous occupations that require colour discrimination. In order to identify valid pre-employment colour vision testing methods, tests actually used and in particular the Ishihara test are considered. After a careful examination we conclude that candidates who pass the Ishihara test have sufficient colour perception to meet ordinary needs and further trade test are not necessary. Nevertheless we do not consider sufficient to rely on the Ishihara test alone to reject a candidate. The diagnostic algorihm proposed is recommended in pre-employment medical examinations for distinguishing people with normal colour vision, slight or sever colour deficiency. PMID- 11386183 TI - [Assessment of problems associated with caregiving: the family strain questionnaire]. AB - Our study concerns a questionnaire for the evaluation of caregiving-related problems: the Family Strain Questionnaire. The questionnaire, which is composed by a structured interview and 47 dychotomous items, was administered to 409 caregivers of patients with different kinds of chronic diseases. Factorial analysis shows a structure of 5 factors named: emotional strain, social involvement, knowledge of the disease, family relationships and thoughts about death. The FSQ is sensible in discriminating different groups of caregivers. This paper presents the psychometric properties as well as the clinical advantages evidenced by the FSQ. PMID- 11386184 TI - [Occupational allergies: magnitude of the problem]. AB - The prevalence of allergic diseases is progressively increasing, but the proportion attributable to occupational agents is not completely known. Most epidemiological data regards occupational asthma due to sensitising agents, which is the most common and frequently compensated occupational respiratory disease. The epidemiology of the so called "irritant asthma" and of occupational rhinitis is less clear, although rhinitis is often associated to asthma and/or precedes its clinical manifestation. The epidemiology of occupational allergic dermatitis has not been completely investigated as well. The present paper is introductory on some selected papers presented in the First Regional Conference on Occupational Allergic Diseases which has been held in Pavia in 1999 with the main goal of giving a picture of the situation of these diseases in our country. PMID- 11386185 TI - [Occupational skin allergies in 1993-1998]. AB - There are few data on the frequency of occupational allergic dermatitis in the context of occupational diseases and there is a need for coordinated large-scale studies to compare the experiences of different Occupational Health Departments involved in the surveillance of allergic occupational diseases. Therefore 6 Occupational Health Departments of Northern Italy (Bergamo, Brescia, Cremona, Desio, Lecco and Milano-CEMOC) decided to bring together all the cases of occupational allergic dermatitis diagnosed between 1993 and 1998. In this report the main characteristics of these 961 allergic dermatitis (888 allergic contact dermatitis, 63 urticaria and 10 airborne contact dermatitis), the possible correlations with gender, age, length of employment, atopy and the occupations at high risk of acquiring allergic dermatitis have been described. PMID- 11386186 TI - [Analysis of occupational allergies reported to the Occupational Medicine and Hygiene Service of the Pavia ASL and to the Hospital Operative Unit of Occupational Medicine in 1993-98]. AB - In this article the cases of occupational allergic diseases notified to 2 Occupational Health Units in Pavia in the period 1993-1998 are reported. A retrospective study was performed to obtain a descriptive analysis of the diseases. In the studied period 58 cases of occupational allergic diseases were notified. This datum probably underestimates the real occurrence of occupational diseases, because of the low accuracy of the notification system. Allergic skin diseases were more frequent (70.7%) than respiratory diseases. PMID- 11386187 TI - [Incremental stress test: comparison between protocols and cardiorespiratory reference values in healthy workers]. AB - The aim of this study was double: to compare two different incremental stress protocols and to obtain reference values for a standardised exercise test in healthy workers. Firstly, eighty healthy male workers, 40 coal miners and 40 hospital workers, aged 19-54, performed in 2 different days 2 cycle ergometer tests up to exhaustion, increasing the work load respectively by 30 watts every 3 minutes (protocol A) and by 30 watts each minute (protocol B). Ventilatory and gas exchange measurements were done by a breath-by-breath apparatus equipped with a turbine and fast gas analysers for O2 and CO2. For each test the ventilatory anaerobic threshold (VAT) was blindly determined as oxygen uptake (V'O2 VAT) using standardised gas exchange and ventilatory indices (V'CO2, V'E, V'E/V'O2) that were found giving comparable results with those derived from the blood lactacte curve. Significant differences were observed between the two protocols only concerning the average work load at exhaustion and at the VAT: the highest being with the protocol B. Conversely, either the maximal oxygen uptake (V'O2 max) and the V'O2 VAT were comparable between protocols, as well as the other cardio-respiratory parameters were at these exercise levels. Oxygen uptake and heart rate increased in average linearly with the work load with very similar slopes in both protocols. Comparable results between protocols were found also as what concerns the slopes of the other physiological variables (V'E, V'CO2) analysed against the V'O2, particularly for exercise levels lower or equal to the individual VAT. Thus, these results suggest a very good comparability between the two protocols, concerning both the levels of maximal and sub-maximal aerobic capacity (V'O2 max, V'O2 VAT) and the cardio-respiratory pattern related to the oxygen uptake. Reference values for the 30 watts/3 minutes cycle ergometer stress test were achieved in other 320 healthy Sardinian workers concerning both the maximal (V'O2 max) and sub-maximal (V'O2 VAT) aerobic capacity and the range of normality for the cardio-respiratory pattern during the test, particularly for completely aerobic work loads, namely work loads not above the V'O2 VAT. These prediction equations can be useful for the evaluation of working capacity of workers employed in manual jobs characterised by moderate-to-high dynamic energy expenditure. PMID- 11386188 TI - [Sampling of allergens in dust deposited in the workplace]. AB - Some workplaces share with domestic dwellings many characteristics favouring house dust mite growth. Moreover it has recently been shown that pets owners can bring allergens to public places with their clothes. So it is possible that significant exposure to indoor allergens can occur outside homes, at the workplace. The recent availability of immunoassays with monoclonal antibodies for indoor allergens has enabled many investigators to quantify exposure to such allergens in epidemiological studies. Analysis of allergens in settled dust is a simple method of quantification exposure to indoor allergens. The concentrations of indoor allergens in public places have already been investigated and high levels of indoor allergens have been reported. A study performed by our group in offices (banks and media) in different regions of Italy has also shown significant levels of indoor allergens. Thus, evaluating exposure to indoor allergens at the workplace is critical to evaluate risk factors for sensitization and elicitation of symptoms in sensitized subjects and such data help in addressing correctly the problem of reducing exposure levels. PMID- 11386189 TI - [Sampling of airborne allergens: practical experience]. AB - The availability of specific monoclonal and policlonal antibodies and the standardization of allergen extracts enables to quantify the exposure to airborne allergens in the domestic environment and at the workplace. In this article we report our experience coming from three studies based upon the measurement of airborne allergens in different environments. In a pharmaceutical factory we measured levels of laboratory animals allergens and evaluated the prevalence of sensitization to the same allergens among the workers. Airborne latex allergens and latex allergens content in latex gloves were measured in hospitals. In bakeries we evaluated the prevalence of sensitization to flour and measured flour allergens levels. The results of our studies give information useful to improve methods of prevention, diagnosis, and therapy of occupational allergic diseases. PMID- 11386190 TI - [Exposure to latex in the health setting: practical experience]. AB - Latex allergy is progressively increasing due to the widespread use of individual protective disposals, especially latex gloves among health care workers. A multidisciplinary Working Group of the Association of Health Care Workers' Preventive Medicine published, in 1995, the Guidelines for the prevention of allergic reactions both in workers and patients. The Group identified a diagnostic protocol, including the assessment of predisposition, cross-reactions, level of exposure and symptoms; etiological diagnosis needs specific, allergologic and provocative tests. Primary prevention, pre-employment education and work restrictions for sensitised workers were particularly taken into account. A short questionnaire was prepared to identify latex sensitisation in patients undergoing invasive procedures; tests are performed only in selected subjects. Preventive measures require the use of nonlatex gloves and the identification and substitution of all latex products in health care settings; such measures are necessary when emergency procedures are performed. PMID- 11386191 TI - [Respiratory occupational allergies: the experience of the Hospital Operative Unit of Occupational Medicine in Lombardy from 1990 to 1998]. AB - A retrospective study on occupational rhinitis and asthma diagnosed in 7 occupational health institutes in Lombardia (North-West Italy) was performed using a standardized card. 141 cases of rhinitis and 281 cases of asthma due to sensitization to occupational agents were analyzed and their clinical characteristics, aetiology, diagnostic methods and associated allergic diseases were determined. In this population the most frequent agents of occupational rhinitis were wheat flour and latex, whereas those of occupational asthma were latex and isocyanate. More than half of the subjects had more than one clinical manifestation of allergy. In 92 out of the 281 asthmatic patients rhinitis was the first clinical manifestation, particularly in subjects sensitized to high molecular weight substances, and preceded, asthma by 12 months as a mean. Specific bronchial provocation tests were useful for the diagnosis of asthma in 153 of the asthmatic patients and 45 of them had an isolated late bronchial reaction following the specific stimulus. At diagnosis 61 subjects (21.7%) had FEV1 < 80% of predicted; factors associated to ventilatory impairment were sensitization to high molecular weight substances, duration of exposure to the sensitizing agent, persistence of exposure after onset of symptoms. PMID- 11386192 TI - [Report on the activity of the Lombardy Association of Occupational Medicine and Industrial Hygiene (ALMLII) in 1998-2000]. PMID- 11386193 TI - Where would we be without the egg? A conference about nature's original functional food. Amelia Island, Florida, USA. February 25-27, 2000. PMID- 11386194 TI - Benefits of parental presence outweigh risks. PMID- 11386195 TI - [Japan Surgical Society 101st annual meeting. Sendai, Japan, April 11-13, 2001. Abstracts]. PMID- 11386196 TI - Proceedings of the 2nd International Conference on Food Factors. December 12-17, 1999. Kyoto, Japan. PMID- 11386197 TI - Heart 2000. Proceedings of the VII National Congress of the Italian Society of Cardiovascular Research. Bologna, Italy, September 20-22, 2000. PMID- 11386198 TI - 1st Educational Meeting in Cardiovascular Medicine. Bologna, Italy, September 22 23, 2000. Abstracts. PMID- 11386199 TI - The Importance of the Child Health Services to the Health of Children. Proceedings of the conference on Child Health Services in Sigtuna, Sweden, 23-25 September 1999. PMID- 11386200 TI - The vascular endothelium: basic and clinical aspects. Advances in management of malignancies. Abstracts of the 3rd and 6th International Congress. Pisa, Italy, October 2-4, 2000. PMID- 11386201 TI - Dreaming and waking consciousness: a character recognition study. AB - The formal features of dream characters were studied in a sample of 320 dream reports submitted by 33 adult subjects (13 male, 20 female) of varying ages in a university extension course. Subjects were queried by questionnaire about dream characters immediately after recording their dreams upon awakening in their normal home setting. It was found that 48% of characters represented a named personage known to the dreamer, 35% were generically identified by their social role (e.g., policeman) or abstract relation to the dreamer (e.g., a friend) while only 16% were wholly novel. Seventy-seven percent of characters were pseudosensorily present in the dream whereas 23% were present only by mention or thought. Subjects were allowed to endorse one or more of four bases of recognition and, among named characters, 32% were identified by 'appearance', 21% by 'behavior', 45% by 'face', and 44% by 'just knowing' (with the respective percentages for generic characters being 39%, 38%, 9% and 40%). Fourteen percent of named and generic characters had associated some element of bizarreness most frequently consisting of an incongruous feature. Comparing the 25 longest and 25 shortest reports, named subjects were significantly more common in the shortest reports whereas generic and unknown characters were more common in the longest reports. Results are interpreted in neurocognitive terms as possibly reflecting a decrease during dreaming relative to waking in the exchange of information between inferotemporal face identification areas and prefrontal areas subserving logic and working memory. PMID- 11386202 TI - Optimization of sigma amplitude threshold in sleep spindle detection. AB - Sleep spindles are transient EEG waveforms of non-rapid eye movement sleep. There is considerable intersubject variability in spindle amplitudes. The problem in automatic spindle detection has been that, despite this fact, a fixed amplitude threshold has been used. Selection of the spindle detection threshold value is critical with respect to the sensitivity of spindle detection. In this study a method was developed to estimate the optimal recording-specific threshold value for each all-night recording without any visual scorings. The performance of the proposed method was validated using four test recordings each having a very different number of visually scored spindles. The optimal threshold values for the test recordings could be estimated well. The presented method seems very promising in providing information about sleep spindle amplitudes of individual all-night recordings. PMID- 11386203 TI - The effect of sleep fragmentation on cognitive processing using computerized topographic brain mapping. AB - Topographic brain mapping of evoked potentials can be used to localize abnormalities of cortical function. We evaluated the effect of sleep fragmentation on brain function by measuring the visual P300 waveform using brain mapping. Eight normal subjects (Epworth Score +/- SD: 5 +/- 3) underwent tone induced sleep fragmentation and undisturbed study nights in a randomized cross over design. Study nights were followed by topographic brain mapping using a visual information processing test and concurrent event-related potentials. Experimental sleep fragmentation did not significantly increase objective daytime sleepiness or lower cognitive performance on a battery of cognitive function tests (all P > or = 0.1). There were no significant topographical delays in P300 latencies with sleep fragmentation (all P > 0.15). However, at sites Fz, F4, T3, C3, Cz and C4 the P300 amplitudes were reduced significantly after sleep fragmentation (all P < 0.05). A reduction in P300 amplitude has previously been interpreted as a decrease in attention. These reductions in P300 amplitudes with sleep fragmentation in frontal, central and temporal brain areas suggest that sleep fragmentation may cause a broad decrease in attention. Sleep fragmentation did not delay P300 latencies in any brain area, and so does not explain the delay in P300 latencies reported in sleep apnoeics. PMID- 11386204 TI - Illumination levels in nursing home patients: effects on sleep and activity rhythms. AB - Studies examining levels of illumination in adult populations have demonstrated that the level and amount of light exposure are lower in the elderly compared with younger adults, particularly in institutionalized patients with dementia. Although insufficient light exposure has been implied as a cause of sleep fragmentation, evidence for such a relationship is scant. Sixty-six institutionalized elderly had their activity and light exposure monitored for a 3 day period. Mean and median light levels, minutes spent over 1000 and over 2000 lux, percent sleep and wake, and number of naps were computed for daytime intervals, defined as 07.00-18.59. Percentages of sleep and wake, number of awakenings and mean duration of wake periods were computed for night-time intervals, defined as 22.00-05.59. Mesor, amplitude and acrophase of activity and of light were determined by cosinor analysis. A mixed linear model was used to assess the effects of daytime Actillume measures on subsequent night-time measures, and vice versa. Spearman correlations were computed, and multiple regression analyses were carried out with light variables and dementia level as predictors and sleep-wake and activity measures as dependent variables. The median light level was 54 lux and a median of only 10.5 min were spent over 1000 lux. Higher light levels predicted fewer night-time awakenings, and severe dementia predicted more daytime sleep and lower mean activity. Increased bright light exposure predicted later activity acrophase. There was an association between the acrophases of light and of activity, with maximum illumination preceding peak activity. These results suggest that daytime light exposure has an impact on both night-time sleep consolidation and timing of peak activity level. PMID- 11386205 TI - Prevalence of sleepiness and its relation to autonomic evidence of arousals and increased inspiratory effort in a community based population of men and women. AB - Degrees of sleep apnoea and daytime sleepiness are quite common in community populations. However the relationship between the two is poor, although sleepiness does correlate better with a history of snoring. It has been suggested that sleep can be fragmented by upper airways obstructive events, short of full apnoeas or hypopnoeas, and that these events may not provoke full cortical arousal, but be detectable through activation of the autonomic system. Failure to detect both these could mask a relationship between 'sleep apnoea' and daytime sleepiness. We have therefore measured sleepiness (Epworth Sleepiness Scale) in addition to both autonomic 'arousals' and inspiratory effort (using pulse transit time) in 473 men and women at home. Although sleepiness was related to a history of snoring, it was not significantly predicted by the measures of autonomic 'arousal', or inspiratory effort. Reported snoring and objectively measured snoring correlated poorly. As in other studies, nocturnal hypoxic dips were correlated with obesity, age, alcohol consumption, drug usage and a history of snoring. These data make it unlikely that sleep fragmentation from subtle variants of sleep apnoea and 'autonomic' (or 'subcortical') arousals are an important source of daytime sleepiness in the community. PMID- 11386206 TI - Tongue protrusion strength and fatiguability: relationship to apnoea/hypopnoea index and age. AB - The sleep apnoea/hypopnoea syndrome (SAHS) is characterized by retroglossal or retropalatal narrowing. The site of obstruction, and the fact that negative pressure in the upper airway increases retroglossal airway size, suggests that tongue muscles may play a role in the maintenance of upper airway patency. We therefore hypothesized that tongue protrusion strength and fatiguability may be predictors of apnoea/hypopnoea index, vary with age and may be different in SAHS patients and normal subjects. Maximal strength (Fmax) and fatiguability (measured as the total time subjects were able to maintain 50% Fmax on three consecutive occasions separated by 30 s) were assessed using a force transducer in 98 consecutive apnoeic/hypopnoeic male patients referred to our laboratory for sleep studies [apnoea/hypopnoea index (AHI) range 3-130/h, age range 30-74 y]. Fmax and fatiguability were also compared in 15 male SAHS patients (mean AHI 20/h) and 15 nonsnoring male subjects matched for age, body mass index and fat free mass. A further 26 SAHS patients had tongue protrusion strength/fatiguability measured before, during and after a night's sleep. Log AHI was only weakly correlated with Fmax (r=- 0.21; P=0.03) and age (r=0.23; P=0.025), but not to fatiguability (P > 0.05). Comparison between SAHS and nonsnoring subjects did not demonstrate significant differences in Fmax (P=0.1) or fatiguability (P=0.1). There was no evidence of a change in muscle strength (P > 0.05) or fatigue (P > 0.05) during the course of a night's sleep. We conclude that tongue protrusion strength and fatiguability are unlikely to be important factors in the pathogenesis of SAHS. PMID- 11386207 TI - Differentiating sleep continuity effects from sleep stage effects. PMID- 11386208 TI - Proceedings of the 5th International Congress of Comparative Physiology and Biochemistry. Calgary, Alberta, Canada. August 1999. PMID- 11386209 TI - Medicine in Split--historic background. PMID- 11386210 TI - [Epidemiological determinants of skin photoageing in a general adult population in France]. PMID- 11386211 TI - [Environmental and hereditary factors in the causation of cancer]. PMID- 11386212 TI - [Paternal exposure to dioxin and sex ratio in the offspring]. PMID- 11386213 TI - Helicobacter pylori pathogenicity: the bacterium and the host. PMID- 11386214 TI - Non-ulcer dyspepsia and patient management, including diagnosis and treatment. PMID- 11386215 TI - Helicobacter pylori and other Helicobacter spp. chronic infections and extragastric diseases. PMID- 11386216 TI - Proceedings of the Surgical Infection Society Meeting. Providence, Rhode Island, USA. April 27-29, 2000. PMID- 11386218 TI - Quality management in head injury care. Abstracts of the 4th Congress of the Euroacademy for multidisciplinary Neurotraumatology. Tenerife, Spain, 2-4 February 1999. PMID- 11386217 TI - Interventions to Prevent HIV Risk Behaviors. NIH Consensus Development Conference. February 11-13, 1997. PMID- 11386219 TI - Human genomics: The basis of the medicine of tomorrow. Validating and using pharmacogenomics. Proceedings of the 2nd IFCC-Roche Diagnostics Conference. Kyoto, Japan, 16-19 April 2000. PMID- 11386220 TI - Proceedings and abstracts of the 17th Annual Symposium on Nonhuman Primate Models for AIDS. October 6-9, 1999. New Orleans, Louisiana, USA. PMID- 11386221 TI - Proceedings of the EC New Genomes Meeting. 23-25 April 1999. PMID- 11386222 TI - A polymorphic microsatellite located within the second intron of the methylmalonyl-CoA mutase (MUT) gene on SSC 1. PMID- 11386223 TI - [Drug resistance in Mycobacterium tuberculosis. A multicenter study of the Barcelona area. Grupo de Trabajo sobre Resistencias en Tuberculosis]. AB - BACKGROUND: The aims of this multicenter study was to establish the level of primary and acquired drug resistance of M. Tuberculosis strains isolated in Barcelona and to identify possible risk groups using clinical data. PATIENTS AND METHODS: All tuberculosis patients with isolation and identification of M. tuberculosis strains from October 1995 to September 1997 were included. Susceptibility tests isoniazid, rifampin, ethambutol, streptomycin and pyrazinamide were performed using the Bactec 460 system and the proportions method on solid medium. Logistic progression was used for statistical analysis. RESULTS: The total number of patients included was 1,749 (1,535 non-treated and 214 previously treated). Primary drug resistance was 5.7% (isoniazid 3.8%; rifampin 1.0%, streptomycin 2.1%, ethambutol 0.3% and pyrazinamide 1.0%). Acquired drug resistance was 20.5% (isoniazid 17.3%, rifampin 9.8%, ethambutol 1.9%, streptomycin 4.7% and pyrazinamide 6.5%). Primary drug resistance was associated with people over 60 years old and women. CONCLUSIONS: The low level of drug resistance enables antituberculosis treatment of non-treated patients to start with the standardised three-drug regimes except in the case of foreign people from countries with a high level of drug resistance. Susceptibility tests are recommended on all M. tuberculosis strains isolated, together with controlled studies of drug resistance surveillance. PMID- 11386224 TI - Auditory language comprehension: an event-related fMRI study on the processing of syntactic and lexical information. AB - The functional specificity of different brain regions recruited in auditory language processing was investigated by means of event-related functional magnetic resonance imaging (fMRI) while subjects listened to speech input varying in the presence or absence of semantic and syntactic information. There were two sentence conditions containing syntactic structure, i.e., normal speech (consisting of function and content words), syntactic speech (consisting of function words and pseudowords), and two word-list conditions, i.e., real words and pseudowords. The processing of auditory language, in general, correlates with significant activation in the primary auditory cortices and in adjacent compartments of the superior temporal gyrus bilaterally. Processing of normal speech appeared to have a special status, as no frontal activation was observed in this case but was seen in the other three conditions. This difference may point toward a certain automaticity of the linguistic processes used during normal speech comprehension. When considering the three other conditions, we found that these were correlated with activation in both left and right frontal cortices. An increase of activation in the planum polare bilaterally and in the deep portion of the left frontal operculum was found exclusively when syntactic processes were in focus. Thus, the present data may be taken to suggest an involvement of the left frontal and bilateral temporal cortex when processing syntactic information during comprehension. PMID- 11386225 TI - Local defense mechanisms in ENT. 21 October 2000. Proceedings. PMID- 11386226 TI - Proceedings of the 5th International Conference on Circulatory Support Devices for Severe Cardiac Failure. New York, New York, USA. September 15-17, 2000. PMID- 11386227 TI - Methods on in situ hybridization, immunohistochemistry and beta-galactosidase reporter gene detection. PMID- 11386229 TI - Proceedings of the 2nd International Symposium on Cerebral and Spinal Cord AVM. 13-14 June 1998, Kyoto, Japan. PMID- 11386228 TI - Proceedings of the 11th Asian Pacific Congress of Gastroenterology and the 8th Asian Pacific Congress of Digestive Endoscopy. 10-14 March 2000, Hong Kong, China. PMID- 11386230 TI - [NSAID-induced necrotizing fasciitis]. PMID- 11386231 TI - [Italian Society of Medical Radiology list of members as of 31 December 2000]. PMID- 11386232 TI - Two similar averages for respiratory muscle activity. PMID- 11386233 TI - Ventilation in mice with a solenoid-controlled airflow. PMID- 11386234 TI - 22nd Annual Meeting of the Society for Clinical Trials. Denver, Colorado, USA. May 20-23, 2001. Abstracts. PMID- 11386236 TI - German Society for Experimental and Clinical Pharmacology and Toxicology 42nd spring meeting. 13-15 March 2001, Mainz, Germany. Abstracts. PMID- 11386235 TI - Japanese Society of Plant Physiologists annual meeting. Fukuoka, Japan, March 23 26, 2001. Abstracts. PMID- 11386237 TI - Honor the pioneers. PMID- 11386238 TI - The effect of tamoxifen therapy on serum apolipoprotein A-I level is modified by the apolipoprotein E phenotype. PMID- 11386239 TI - International Association for Dental Research 79th general session. June 27-30, 2001. Chiba, Japan. Abstracts. PMID- 11386240 TI - Humanitarian health tour in Peru: a first-hand account. PMID- 11386241 TI - A look back: early ligatures. PMID- 11386242 TI - Web options for certification review and nursing degree completion debut. On-line nurse practitioner review course. PMID- 11386245 TI - Toxic tanneries. PMID- 11386244 TI - Full disclosure in Europe. PMID- 11386243 TI - No ability to produce tetrodotoxin in bacteria. PMID- 11386246 TI - The Physiological Society proceedings of the scientific meeting held at King's College London, 18-20 December 2000. PMID- 11386247 TI - American Academy of Neurology 53rd annual meeting. May 5-11, 2001, Philadelphia, Pennsylvania, USA. Abstracts. PMID- 11386248 TI - Immunity and Prevention of Herpes Zoster. Proceedings of an international conference. Osaka, Japan, March 8-10, 1999. PMID- 11386249 TI - 'Slim is not in' among adolescents in Trinidad. PMID- 11386250 TI - American Urological Association annual meeting. June 2-7, 2001. Anaheim, California, USA. Abstracts. PMID- 11386251 TI - 11th Congress of the Division on Experimental Cancer Research (AEK) of the German Cancer Society. 4-6 April 2001, Heidelberg, Germany. Abstracts. PMID- 11386252 TI - Digestive Disease Week and the 102nd Annual Meeting of the American Gastroenterological Association. May 20-23, 2001. Atlanta, Georgia, USA. Abstracts. PMID- 11386253 TI - Markers of joint destruction: principles, problems, and potential. PMID- 11386254 TI - Periarticular bone mineral density at the knee joint. PMID- 11386255 TI - A mnemonic for SLE diagnostic criteria. PMID- 11386256 TI - Can rheumatologists agree on a diagnosis of inflammatory arthritis in an early synovitis clinic? PMID- 11386257 TI - Ultrasound guided injection of plantar fasciitis. PMID- 11386259 TI - From the Centers for Disease Control and Prevention. Fatal occupational injuries- United States, 1980-1997. PMID- 11386258 TI - HLA-DRB1 and DQB1 genes in anticentromere antibody positive patients with SSc and primary biliary cirrhosis. PMID- 11386260 TI - From the Centers for Disease Control and Prevention. Baler and compactor-related deaths in the workplace--United States, 1992-2000. PMID- 11386261 TI - From the Centers for Disease Control and Prevention. Nonfatal occupational injuries and illnesses treated in hospital emergency departments--United States, 1998. PMID- 11386262 TI - JAMA patient page. Cholesterol and atherosclerosis. PMID- 11386263 TI - Effect of carvedilol on survival in severe chronic heart failure. AB - BACKGROUND: Beta-blocking agents reduce the risk of hospitalization and death in patients with mild-to-moderate heart failure, but little is known about their effects in severe heart failure. METHODS: We evaluated 2289 patients who had symptoms of heart failure at rest or on minimal exertion, who were clinically euvolemic, and who had an ejection fraction of less than 25 percent. In a double blind fashion, we randomly assigned 1133 patients to placebo and 1156 patients to treatment with carvedilol for a mean period of 10.4 months, during which standard therapy for heart failure was continued. Patients who required intensive care, had marked fluid retention, or were receiving intravenous vasodilators or positive inotropic drugs were excluded. RESULTS: There were 190 deaths in the placebo group and 130 deaths in the carvedilol group. This difference reflected a 35 percent decrease in the risk of death with carvedilol (95 percent confidence interval, 19 to 48 percent; P=0.00013, unadjusted; P=0.0014, adjusted for interim analyses). A total of 507 patients died or were hospitalized in the placebo group, as compared with 425 in the carvedilol group. This difference reflected a 24 percent decrease in the combined risk of death or hospitalization with carvedilol (95 percent confidence interval, 13 to 33 percent; P<0.001). The favorable effects on both end points were seen consistently in all the subgroups we examined, including patients with a history of recent or recurrent cardiac decompensation. Fewer patients in the carvedilol group than in the placebo group withdrew because of adverse effects or for other reasons (P=0.02). CONCLUSIONS: The previously reported benefits of carvedilol with regard to morbidity and mortality in patients with mild-to-moderate heart failure were also apparent in the patients with severe heart failure who were evaluated in this trial. PMID- 11386264 TI - A trial of the beta-blocker bucindolol in patients with advanced chronic heart failure. AB - BACKGROUND: Although beta-adrenergic-receptor antagonists reduce morbidity and mortality in patients with mild-to-moderate chronic heart failure, their effect on survival in patients with more advanced heart failure is unknown. METHODS: A total of 2708 patients with heart failure designated as New York Heart Association (NYHA) functional class III (in 92 percent of the patients) or IV (in 8 percent) and a left ventricular ejection fraction of 35 percent or lower were randomly assigned to double-blind treatment with either bucindolol (1354 patients) or placebo (1354 patients) and followed for the primary end point of death from any cause. RESULTS: The data and safety monitoring board recommended stopping the trial after the seventh interim analysis. At that time, there was no significant difference in mortality between the two groups (unadjusted P=0.16). The results presented here are based on complete follow-up at the time of study termination (average, 2.0 years). There were a total of 449 deaths in the placebo group (33 percent) and 411 deaths in the bucindolol group (30 percent; adjusted P=0.13). The risk of the secondary end point of death from cardiovascular causes was lower in the bucindolol group (hazard ratio, 0.86; 95 percent confidence interval, 0.74 to 0.99), as was the risk of heart transplantation or death (hazard ratio, 0.87; 95 percent confidence interval, 0.77 to 0.99). CONCLUSIONS: In a demographically diverse group of patients with NYHA class III and IV heart failure, bucindolol resulted in no significant overall survival benefit. PMID- 11386265 TI - Effect of a single amino acid change in MHC class I molecules on the rate of progression to AIDS. AB - BACKGROUND: From studies of genetic polymorphisms and the rate of progression from human immunodeficiency virus type 1 (HIV-1) infection to the acquired immunodeficiency syndrome (AIDS), it appears that the strongest susceptibility is conferred by the major-histocompatibility-complex (MHC) class I type HLA B*35,Cw*04 allele. However, cytotoxic T-lymphocyte responses have been observed against HIV-1 epitopes presented by HLA-B*3501, the most common HLA-B*35 subtype. We examined subtypes of HLA-B*35 in five cohorts and analyzed the relation of structural differences between HLA-B*35 subtypes to the risk of progression to AIDS. METHODS: Genotyping of HLA class I loci was performed for 850 patients who seroconverted and had known dates of HIV-1 infection. Survival analyses with respect to the rate of progression to AIDS were performed to identify the effects of closely related HLA-B*35 subtypes with different peptide-binding specificities. RESULTS: HLA-B*35 subtypes were divided into two groups according to peptide-binding specificity: the HLA-B*35-PY group, which consists primarily of HLA-B*3501 and binds epitopes with proline in position 2 and tyrosine in position 9; and the more broadly reactive HLA-B*35-Px group, which also binds epitopes with proline in position 2 but can bind several different amino acids (not including tyrosine) in position 9. The influence of HLA-B*35 in accelerating progression to AIDS was completely attributable to HLA-B*35-Px alleles, some of which differ from HLA-B*35-PY alleles by only one amino acid residue. CONCLUSIONS: This analysis shows that, in patients with HIV-1 infection, a single amino acid change in HLA molecules has a substantial effect on the rate of progression to AIDS. The different consequences of HLA-B*35-PY and HLA-B*35-Px in terms of disease progression highlight the importance of the epitope specificities of closely related class I molecules in the immune defense against HIV-1. PMID- 11386266 TI - Laryngeal transplantation and 40-month follow-up. PMID- 11386267 TI - Images in clinical medicine. Infiltration of bone marrow by a signet-ring-cell gastric carcinoma. PMID- 11386268 TI - Expandable metal stents for the treatment of cancerous obstruction of the gastrointestinal tract. PMID- 11386269 TI - Amyotrophic lateral sclerosis. PMID- 11386270 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 17-2001. A 42-year-old man with multiple pulmonary cysts and recurrent respiratory infections. PMID- 11386272 TI - Transplantation of the larynx--a case report that speaks for itself. PMID- 11386271 TI - Expanding indications for beta-blockers in heart failure. PMID- 11386273 TI - Dietary sodium and blood pressure. PMID- 11386274 TI - Dietary sodium and blood pressure. PMID- 11386275 TI - Dietary sodium and blood pressure. PMID- 11386276 TI - Dietary sodium and blood pressure. PMID- 11386277 TI - A pneumococcal conjugate vaccine and acute otitis media. PMID- 11386278 TI - A pneumococcal conjugate vaccine and acute otitis media. PMID- 11386279 TI - A pneumococcal conjugate vaccine and acute otitis media. PMID- 11386280 TI - A pneumococcal conjugate vaccine and acute otitis media. PMID- 11386281 TI - A pneumococcal conjugate vaccine and acute otitis media. PMID- 11386282 TI - The effect of risedronate on the risk of hip fracture in elderly women. PMID- 11386283 TI - The effect of risedronate on the risk of hip fracture in elderly women. PMID- 11386284 TI - The effect of risedronate on the risk of hip fracture in elderly women. PMID- 11386285 TI - Treatment of calcium-channel-blocker intoxication with insulin infusion. PMID- 11386286 TI - Testing poor pregnant women for cocaine--physicians as police investigators. PMID- 11386287 TI - Annual scientific meeting of the British Society for Haematology. Harrogate, United Kingdom, 23-26 April 2001. Abstracts. PMID- 11386288 TI - [Areas of family counseling]. PMID- 11386290 TI - [82nd German Radiology Congress. 23-26 May 2001, Wiesbaden, Germany. Abstracts]. PMID- 11386289 TI - Association of Surgeons of Great Britain and Ireland and Surgical Research Society annual meeting. Birmingham, 25-27 April 2001. Abstracts. PMID- 11386291 TI - Concern about records of fibrosing colonopathy study. PMID- 11386292 TI - Sexually transmitted diseases in HIV-1-infected patients. PMID- 11386293 TI - Sexually transmitted diseases in HIV-1-infected patients. PMID- 11386294 TI - Prevention of pre-eclampsia. PMID- 11386295 TI - Learn from our errors. PMID- 11386296 TI - Exit of platelet glycoprotein-IIb/IIIa-receptor inhibitors? PMID- 11386297 TI - Exit of platelet glycoprotein-IIb/IIIa-receptor inhibitors? PMID- 11386299 TI - A life worth living. PMID- 11386298 TI - Informed consent in cancer registries. PMID- 11386301 TI - Anaerobic batch degradation of solid poultry slaughterhouse waste. AB - We studied anaerobic batch degradation of solid poultry slaughterhouse wastes with different initial waste and inoculum concentrations and waste-to-inoculum ratios and simulated the dynamics of the process with a new generation model. Our modelling results suggest that inhibited propionate degradation by long-chain fatty acids (LCFA) and inhibited hydrolysis by a high propionate concentration constituted the rate-limiting step in the waste degradation. Palmitate was the most abundant LCFA in the assays. Within 27 days of incubation, up to 0.55 to 0.67 m3 of methane (STP)/kg VS added was produced under the studied conditions. Lower waste-to-inoculum ratios exhibited a faster onset and rate of specific methane production. In all the assays, ammonification occurred within 3 to 6 days and accounted for 50 to 60% of total nitrogen. PMID- 11386300 TI - [Relevant aspects of available infrastructure of private medical practice, 1999]. PMID- 11386302 TI - Master index, volumes 465-486 (2000). PMID- 11386303 TI - The rH index for quality and environmental controls: standards and application procedures. AB - The quantity rH, an index of the reducing power of a redox system is, in conceptual and operational terms, coordinatively linked with the two familiar electroanalytical quantities: redox potential (EO/R) and pH, and has recently become important in quality and environmental controls (hydrographic network, human consumption waters, industrial effluents, etc.). It has some significant problems of standardization and methodological applications, especially on passing from pure water medium to aqueous-organic media, and the results of the most recent researches are here presented. PMID- 11386305 TI - Patient notes: living with diabetes. PMID- 11386304 TI - The consequences of the Holocaust on child survivors and children of survivors. PMID- 11386306 TI - Estimating the employment and earnings costs of mental illness: recent developments in the United States. AB - Substantial attention has recently been focused on both the prevalence and consequences of mental illness. Generally, public interest in the costs of mental illness has been limited to the direct costs of treating the mentally ill. In this paper, we consider the magnitude and importance of a major component of the indirect costs of mental illness: employment and earnings losses. We first describe the technical difficulties involved in estimating these costs. We then describe new data and recent advances in the United States that have improved our ability to make such estimates. Our conclusions from the recent research are that each year in the United States 5-6 million workers between the ages of 16 and 54 lose, fail to seek, or cannot find employment as a consequence of mental illness. Among those who do work, we estimate that mental illness decreases annual income by an amount between $3,500 and $6,000. We then discuss an emerging challenge to the traditional method for arriving at such estimates: the friction cost approach. We describe both the conceptual and technical differences between the friction cost method and the traditional human capital approach. We conclude that while economic context has much to do with whether one relies on human capital or friction cost estimates, each can offer useful information about labor market losses due to mental illness. PMID- 11386307 TI - Models of acculturation and health behaviors among Latino immigrants to the US. AB - A basic premise of much of the health research conducted with immigrant groups is that culturally based behaviors change over time as a result of acculturation, i.e., interaction with the mainstream US culture. However, models of acculturation have not taken into account how group-specific characteristics and the varying social and political contexts immigrant groups face may impact the acculturation process. In this study of 150 families, we examined the inter relationship of indicators of acculturation among two Latino groups to discern the impact of gender and country of origin on the relationship between variables. Results indicated that increased years of residence in the United States had the predictable impact of increased competence in English and increased use of English, but had differing impact by country of origin on the cultural orientation of the respondents' environment and on ethnic identification. Also, gender was associated with differing levels of English language use and with perceived social acceptance, such that males used more English and reported less social acceptance than females. Loading separately from the language and cultural behavior variables, this factor, perceived social acceptance, merits research as a predictor of service use given that respondents understood non-acceptance as resulting from being identified as Latino. not from behaving differently from the mainstream. The differing patterns of association by country of origin and by gender and the measurement issues these raise, highlight the importance of specifying more complex models of a cculturation than is done typically in research with Latinos. PMID- 11386308 TI - Hopelessness predicts mortality in older Mexican and European Americans. AB - OBJECTIVE: The purpose of this study was to evaluate the relationship between hopelessness and mortality in a biethnic cohort of older community-dwelling Mexican Americans, the most rapidly growing segment of the elderly, and European Americans. METHODS: A total of 795 persons aged 64 to 79 years completed an English or Spanish version of the 30-item Geriatric Depression Scale on entering the San Antonio Longitudinal Study of Aging, an epidemiologic survey, between 1992 and 1996. Women constituted 58% and Mexican Americans 54% of this randomly selected sample. Subjects who answered "no" to the item "Are you hopeful about the future?" were classified as hopeless. RESULTS: As of August 1999, 29% of the 73 hopeless subjects had died, compared with 11% of the hopeful, a highly significant difference. The mortality rates for cardiovascular disease and cancer were significantly greater among the hopeless subjects (7%) than among the hopeful (3%). Hopelessness predicted all-cause mortality in a Cox proportional hazards model adjusted for age, ethnic background, current smoking status, number of comorbid medical conditions, self-rated health, and frequency of social contacts (risk ratio = 2.23, 95% confidence interval = 1.33 to 3.76, p = .0026). Neither sex nor probable depression was a significant predictor of mortality in this model. CONCLUSION: These findings, together with those of others, suggest that hopelessness is a significant predictor of mortality in older and middle aged adults of various ethnic backgrounds. Further research is needed to evaluate the mechanisms that underlie this phenomenon and the effects of treating hopelessness on the quality and duration of subjects' lives. PMID- 11386309 TI - Glucosamine sulphate and osteoarthritis. PMID- 11386310 TI - Glucosamine sulphate and osteoarthritis. PMID- 11386311 TI - Glucosamine sulphate and osteoarthritis. PMID- 11386312 TI - Glucosamine sulphate and osteoarthritis. PMID- 11386313 TI - Ischaemic colitis and meloxicam. PMID- 11386314 TI - Angiotensin-II-receptor inhibitors in pregnancy. PMID- 11386315 TI - Angiotensin-II-receptor inhibitors in pregnancy. PMID- 11386316 TI - Angiotensin-II-receptor inhibitors in pregnancy. PMID- 11386317 TI - Lung-volume reduction. PMID- 11386319 TI - Hyponatraemia after colonoscopy. PMID- 11386320 TI - Common risk factors in type 1 diabetes and asthma. PMID- 11386321 TI - Osteoporosis after organ transplantation. PMID- 11386322 TI - Osteoporosis after organ transplantation. PMID- 11386323 TI - Osteoporosis after organ transplantation. PMID- 11386325 TI - Trade systems in less-developed countries. PMID- 11386324 TI - Trade systems in less-developed countries. PMID- 11386326 TI - Seizures after buproprion overdose. PMID- 11386327 TI - Fallacies in modern medical education. PMID- 11386328 TI - Paediatric brain death in Japan. PMID- 11386329 TI - Pre-gay, post-queer: Thai perspectives on proliferating gender/sex diversity in Asia. PMID- 11386330 TI - Survival through pluralism: emerging gay communities in the Philippines. PMID- 11386331 TI - Gay and lesbian couples in Malaysia. PMID- 11386332 TI - Let them take ecstasy: class and Jakarta lesbians. PMID- 11386333 TI - Drink, stories, penis, and breasts: lesbian tomboys in Taiwan from the 1960s to the 1990s. PMID- 11386334 TI - Asian values, family values: film, video, and lesbian and gay identities. PMID- 11386335 TI - Homosexual rights as human rights in Indonesia and Australia. PMID- 11386336 TI - Variations on a common theme? Gay and lesbian identity and community in Asia. PMID- 11386337 TI - Homosexuality and the cultural politics of tongzhi in Chinese societies. PMID- 11386338 TI - Becoming a gay activist in contemporary China. PMID- 11386339 TI - Mapping the vicissitudes of homosexual identities in South Korea. PMID- 11386340 TI - Tiptoe out of the closet: the before and after of the increasingly visible gay community in Singapore. PMID- 11386341 TI - Culture, sexualities, and identities: men who have sex with men in India. PMID- 11386342 TI - Initiation of the Streptomyces chromosome replication. PMID- 11386343 TI - Isolation and cloning of Streptomyces terminal fragments. AB - Streptomyces species have a linear chromosome of approximately 8 Mb in size. Many strains also carry linear plasmids. Most of these linear elements contain terminal proteins covalently bound to the 5' ends of the DNA. Using a method for the visualisation of terminal DNA fragments in agarose gels, it was possible to see three fragments in S. rimosus and five fragments in S. avermitilis. The method was also used to clone the 298 bp BamHI fragment carrying the left end of plasmid SLP2. Analysis of the sequence showed that the end resembled other Streptomyces chromosome and plasmid ends, but there were eight palindromes (instead of seven) and a tandem duplication of a 14 bp sequence. PMID- 11386344 TI - Chromosome topology and genome size of selected actinomycetes species. AB - Information about the genome organization of actinomycetes species is restricted to a few genera: Corynebacterium, Mycobacterium, Rhodococcus, Saccharopolyspora and Streptomyces. Streptomyces species and Saccharopolyspora erythraea were shown to contain a single linear 8 Mb chromosome. In contrast, the Corynebacterium, Mycobacterium and Rhodococcus species studied were demonstrated to possess a smaller (3 Mb-6.5 Mb) single circular chromosome. To investigate whether linear chromosome topology and genome sizes above 7 Mb are unique features of Streptomyces and S. erythraea we have started to investigate the chromosome topology, the genome size and the status of accessory elements of additional actinomycetes species: Actinoplanes philippinensis, Amycolatopsis orientalis, Micromonospora chalcea, Nocardia asteroides, Rhodococcus opacus and Streptoverticillium abikoense. Our data which are based on PFGE experiments clearly suggest that large genome sizes and chromosome linearity are seen in mycelium forming actinomycetes genera. In addition we have identified large linear plasmids in Nocardia asteroides, Streptoverticillium abikoense and Rhodococcus opacus. PMID- 11386345 TI - Alteration of substrate specificity of valine dehydrogenase from Streptomyces albus. AB - The catabolism of branched chain amino acids, especially valine, appears to play an important role in furnishing building blocks for macrolide and polyether antibiotic biosyntheses. To determine the active site residues of ValDH, we previously cloned, partially characterized, and identified the active site (lysine) of Streptomyces albus ValDH. Here we report further characterization of S. albus ValDH. The molecular weight of S. albus ValDH was determined to be 38 kDa by SDS-PAGE and 67 kDa by gel filtration chromatography indicating that the enzyme is composed of two identical subunits. Optimal pHs were 10.5 and 8.0 for dehydrogenase activity with valine and for reductive amination activity with alpha-ketoisovaleric acid, respectively. Several chemical reagents, which modify amino-acid side chains, inhibited the enzyme activity. To examine the role played by the residue for enzyme specificity, we constructed mutant ValDH by substituting alanine for glycine at position 124 by site-directed mutagenesis. This residue was chosen because it has been considered to be important for substrate discrimination by phenylalanine dehydrogenase (PheDH) and leucine dehydrogenase (LeuDH). The Ala-124-Gly mutant enzyme displayed lower activities toward aliphatic amino acids, but higher activities toward L-phenylalanine, L tyrosine, and L-methionine compared to the wild type enzyme suggesting that Ala 124 is involved in substrate binding in S. albus ValDH. PMID- 11386346 TI - Sugar uptake and utilisation in Streptomyces coelicolor: a PTS view to the genome. AB - Our research group is studying the phosphotransferase system (PTS) of Streptomyces coelicolor, which, in other bacteria, is centrally involved in carbon source uptake and regulation. We have surveyed the public available S. coelicolor genome sequence produced by the ongoing genome sequencing project for pts gene homologues (http://www.sanger.ac.uk/Projects/S_coelicolor/). Three genes encoding homologues of the general PTS components enzyme I (ptsI), HPr (ptsH), and enzyme IIA(Crr) (crr; IIA(Glc)-homologue) and six genes encoding homologues of sugar-specific PTS components were identified. The deduced primary sequences of the sugar-specific components shared significant similarities to PTS permeases of the mannitol/fructose family and of the glucose/sucrose family. A model is presented, in which possible functions of the novel described PTS homologues are discussed. PMID- 11386347 TI - Glucose kinase of Streptomyces coelicolor A3(2): large-scale purification and biochemical analysis. AB - Glucose kinase of Streptomyces coelicolor A3(2) is essential for glucose utilisation and is required for carbon catabolite repression (CCR) exerted through glucose and other carbon sources. The protein belongs to the ROK-family, which comprises bacterial sugar kinases and regulators. To better understand glucose kinase function, we have monitored the cellular activity and demonstrated that the choice of carbon sources did not significantly change the synthesis and activity of the enzyme. The DNA sequence of the Streptomyces lividans glucose kinase gene glkA was determined. The predicted gene product of 317 amino acids was found to be identical to S. coelicolor glucose kinase, suggesting a similar role for this protein in both organisms. A procedure was developed to produce pure histidine-tagged glucose kinase with a yield of approximately 10 mg/l culture. The protein was stable for several weeks and was used to raise polyclonal antibodies. Purified glucose kinase was used to explore protein protein interaction by surface plasmon resonance. The experiments revealed the existence of a binding activity present in S. coelicolor cell extracts. This indicated that glucose kinase may interact with (an)other factor(s), most likely of protein nature. A possible cross-talk with proteins of the phosphotransferase system, which are involved in carbon catabolite repression in other bacteria, was investigated. PMID- 11386348 TI - Carbon-source-dependent transcriptional control involved in the initiation of cellular differentiation in Streptomyces griseus. AB - Carbon source is one of the environmental factors that affects cellular differentiation of Streptomyces. We have identified the craA gene as a putative negative regulator involved in the carbon-source-dependent initiation of cellular differentiation in Streptomyces griseus. Carbon-source-dependent transcriptional repression of craA, which is caused by binding of a putative repressor protein to its promoter region, is proposed to result in the initiation of aerial mycelium formation. The presence of a craA homologue in the chromosome of Streptomyces coelicolor A3(2) implicates the existence of a similar regulatory mechanism in this organism. The repression of craA-promoter activity in glucose media could be alleviated not only by replacing glucose with maltose but also by supplying copper, which suggests that the stimulatory effect of copper on cellular differentiation in Streptomyces is excerted via abolishment of glucose-repression of craA. PMID- 11386349 TI - Effects of increased and deregulated expression of cell division genes on the morphology and on antibiotic production of streptomycetes. AB - This paper describes the effects of increased expression of the cell division genes ftsZ, ftsQ, and ssgA on the development of both solid- and liquid-grown mycelium of Streptomyces coelicolor and Streptomyces lividans. Over-expression of ftsZ in S. coelicolor M145 inhibited aerial mycelium formation and blocked sporulation. Such deficient sporulation was also observed for the ftsZ mutant. Over-expression of ftsZ also inhibited morphological differentiation in S. lividans 1326, although aerial mycelium formation was less reduced. Furthermore, antibiotic production was increased in both strains, and in particular the otherwise dormant actinorhodin biosynthesis cluster of S. lividans was activated in liquid- and solid-grown cultures. No significant alterations were observed when the gene dosage of ftsQ was increased. Analysis by transmission electron microscopy of an S. coelicolor strain overexpressing ssgA showed that septum formation had strongly increased in comparison to wild-type S. coelicolor, showing that SsgA clearly influences Streptomyces cell division. The morphology of the hyphae was affected such that irregular septa were produced with a significantly wider diameter, thereby forming spore-like compartments. This suggests that ssgA can induce a process similar to submerged sporulation in Streptomyces strains that otherwise fail to do so. A working model is proposed for the regulation of septum formation and of submerged sporulation. PMID- 11386350 TI - Transcriptional and functional analysis of the gene for factor C, an extracellular signal protein involved in cytodifferentiation of Streptomyces griseus. AB - Factor C is an extracellular signal protein involved in cellular differentiation in Streptomyces griseus. Nuclease S1 mapping experiments revealed that transcription of the gene takes place from a single promoter in a developmental stage specific manner. The latter was also confirmed by in vivo promoter probing. The sequence of its promoter suggests that the gene is not transcribed by the major sigma factor. The cloned gene expressed from its own promoter in low- and high-copy-number vectors restored normal sporulation to a bald mutant of Streptomyces griseus. Computer analysis of the amino acid sequence revealed the presence of a transmembrane localization segment with the N-terminus positioned inside the cell. These data fit well into our working model that points at an important role for factor C in the morphogenesis of Streptomyces griseus. PMID- 11386351 TI - Aspects of the biosynthesis of non-aromatic fungal polyketides by iterative polyketide synthases. AB - Lovastatin biosynthesis in Aspergillus terreus involves two unusual type I multifunctional polyketide syntheses (PKSs). Lovastatin nonaketide synthase (LNKS), the product of the lovB gene, is an iterative PKS that interacts with LovC, a putative enoyl reductase, to catalyze the 35 separate reactions in the biosynthesis of dihydromonacolin L, a lovastatin precursor. LNKS also displays Diels-Alderase activity in vitro. Lovastatin diketide synthase (LDKS) made by lovF, in contrast, acts non-iteratively like the bacterial modular PKSs to make (2R)-2-methylbutyric acid. Then, like LNKS, LDKS interacts closely with another protein, the LovD transesterase enzyme that catalyzes attachment of the 2 methylbutyric acid to monacolin J in the final step of the lovastatin pathway. Key features of the genes for these four enzymes and others, plus the regulatory and self-resistance factors involved in lovastatin production, are also described. PMID- 11386352 TI - Streptomycetes in micro-cultures: growth, production of secondary metabolites, and storage and retrieval in the 96-well format. AB - Mycelium-forming Streptomyces strains were grown in one milliliter liquid micro cultures in square deep-well microtiter plates. Growth was evaluated with respect to biomass formation and production of secondary metabolites which were found to be very similar in the micro-cultures, bioreactor, and shake flask cultivations, respectively. Despite repetitive sampling and extensive growth on the walls of the wells, no cross contamination occurred. Furthermore, we successfully employed cold storage at -20 degrees C of spore suspensions (in the 96-well format), directly prepared from cultures grown on agar in the microtitre plate. Cultures were retrieved by replicating aliquots from the frozen spore suspensions. PMID- 11386353 TI - Microbial growth and production kinetics of Streptomyces antibioticus Tu 6040. AB - Streptomyces antibioticus Tu 6040 is the producer of simocyclinones, which belong to a novel family of angucyclinone antibiotics some of which show antitumor activities. Growth and antibiotic production is dependent on the medium composition, especially on the carbon and nitrogen source, and on the fermentation conditions. The best results with respect to antibiotic productivity were achieved using a chemically defined medium with glycerol and L-lysine as carbon and nitrogen source, respectively, in an airlift fermenter with minimised shear stress at low gas flow rates withour oxygen limitation. These conditions led to a homogeneous formation of pellets of 1-2 mm in diameter and guaranteed reproducible product yields of the main compound, simocyclinone D8, in the range of 300 mg/l. PMID- 11386354 TI - Molecular detection of bacterial and streptomycete chitinases in the environment. AB - Sets of PCR primers were designed to amplify bacterial chitinases at different levels of specificity. The bacterial chitinase group primers were successful in targeting enzymes classified within the group A glycosyl hydrolases of family 18. The widespread occurrence of group A bacterial chitinases in actinomycetes was demonstrated. Streptomycete chitinase specific primers were designed and a collection of type strains of species changed in the genes Streptomyces were screened and shown to have at least one and usually multiple chitinase genes. The presence of the gene for the chitin binding protein was also demonstrated within the streptomycete type strains. These data indicate that streptomycetes are well equipped to degrade chitin. The detection of group A chitinases in total community DNA is described and a sandy soil shown to contain more than 10 different genes using DGGE to indicate genetic diversity. PMID- 11386355 TI - Occurrence and diversity of plasmids in populations of streptomycetes in soil. AB - Studies were made of naturally occurring plasmids hosted in Streptomyces strains isolated from two different terrestrial ecosystems: an agricultural field and a protected forest area. Six out of the 147 screened isolates contained plasmids. The strains containing these plasmids were all isolated from the agricultural soil. Plasmids were not found among the strains isolated from the forest area. Cross hybridization of the six newly isolated plasmids revealed very high similarities between four of them. However, no similarities were found between the six newly isolated plasmids and well studied streptomycete plasmids such as pIJ101 and SCP2*. The host strains of the four similar plasmids belonged to three different species S. anulatus, S. rochei and S. diastaticus. This implies a possible conjugative transfer of these plasmids within the streptomycete population in the agricultural area. The reason for the absence of streptomycete plasmids from the populations derived from the forest area is discussed. PMID- 11386356 TI - Genomic and phenomic differentiation of Rhodococcus equi and related strains. AB - 16S rDNA sequence and pyrolysis mass spectrometric analyses were carried out on representatives of Rhodococcus equi and marker strains of genera that encompass mycolic acid containing actinomycetes. The R. equi strains formed a monophyletic clade within the evolutionary radiation occupied by members of the genera Nocardia and Rhodococcus. The 16S rDNA sequence data also showed R. equi to be an heterogeneous taxon. This heterogeneity was underscored by the pyrolysis mass spectrometric data. These observations are in line with those of previous studies where similar profiles of relatedness were found between pyrolysis mass spectral data and the results of DNA:DNA pairing and numerical phenetic studies. PMID- 11386357 TI - The diversity of extradiol dioxygenase (edo) genes in cresol degrading rhodococci from a creosote-contaminated site that express a wide range of degradative abilities. AB - Analysis of the bacterial population of soil surface samples from a creosote contaminated site showed that up to 50% of the culturable micro-organisms detected were able to utilise a mixture of cresols. From fifty different microbial isolates fourteen that could utilise more than one cresol isomer were selected and identified by 16S rRNA analysis. Eight isolates were Rhodococcus strains and six were Pseudomonas strains. In general, the Rhodococcus strains exhibited a broader growth substrate range than the Pseudomonas strains. The distribution of various extradiol dioxygenase (edo) genes, previously associated with aromatic compound degradation in rhodococci, was determined for the Rhodococcus strains by PCR detection and Southern-blot hybridization. One strain, Rhodococcus sp. II exhibited the broadest growth substrate range and possessed five different edo genes. Gene disruption experiments indicated that two genes (edoC and edoD) were associated with isopropylbenzene and naphthalene catabolism respectively. The other Rhodococcus strains also possessed some of the edo genes and one (edoB) was present in all of the Rhodococcus strains analysed. None of the rhodococcal edo genes analysed were present in the Pseudomonas strains isolated from the site. It was concluded that individual strains of Rhodococcus possess a wide degradative ability and may be very important in the degradation of complex mixtures of substrates found in creosote. PMID- 11386358 TI - Selective isolation and characterisation of members of the Streptomyces violaceusniger clade associated with the roots of Paraserianthes falcataria. AB - Large numbers of putatively novel streptomycetes were isolated from environmental samples collected from in and around the root system of the tropical angiosperm, Paraserianthes falcataria. Representative isolates were assigned to 37 multi membered and 107 single membered colour groups based on their ability to form pigments on oatmeal and peptone yeast extract iron agars. The largest taxon, colour group 3, encompassed 94 isolates which had morphological properties typical of members of the Streptomyces violaceusniger clade. Twelve representatives of this taxon chosen on the basis of Curie-point pyrolysis mass spectrometric data were compared with representatives of the validly described species which constitute the Streptomyces violaceusniger clade. Six out of the twelve representative strains were readily distinguished from one another and from the marker strains using a combination of genotypic and phenotypic properties. These organisms were consequently considered to merit species status as Streptomyces asiaticus sp. nov., Streptomyces cangkringensis sp. nov., Streptomyces indonesiensis sp. nov., Streptomycesjavensis sp. nov., Streptomyces rhizosphaerius sp. nov. and Streptomyces yogyakartensis sp. nov. PMID- 11386359 TI - The Nocardia salmonicida clade, including descriptions of Nocardia cummidelens sp. nov., Nocardia fluminea sp. nov. and Nocardia soli sp. nov. AB - Large numbers of strains selectively isolated from soil, water and deteriorating vulcanised natural rubber pipe rings were provisionally assigned to the genus Nocardia. Twenty-eight representative isolates were found to have chemical and morphological properties typical of nocardiae. These organisms formed a monophyletic clade in the 16S rDNA tree together with Nocardia salmonicida. Three of the strains, isolates S1, W30 and R89, were distinguished from one another and from representatives of the validly described species of Nocardia using genotypic and phenotypic data. These organisms were considered to merit species status and were named Nocardia cummidelens sp. nov., Nocardia fluminea sp. nov. and Nocardia soli sp. nov. respectively. Additional comparative studies are needed to resolve the finer taxonomic relationships of the remaining isolates assigned to the Nocardia salmonicida clade and to further unravel the extent of nocardial diversity in artificial and natural ecosystems. PMID- 11386360 TI - Teicoplanin biosynthesis genes in Actinoplanes teichomyceticus. AB - The genetic determinants for the biosynthesis of the glycopeptide antibiotic teicoplanin were identified. In order to isolate the corresponding gene cluster, oligonucleotides derived from highly conserved motifs in peptide synthetases were used. These synthetic probes, and gene fragments derived from the balhimycin gene cluster of Amycolatopsis mediterranei, led to the identification of the likely teicoplanin gene cluster centered on a region of ca. 110 kb from the genome of Actinoplanes teichomyceticus, the teicoplanin producer. Partial nucleotide sequences identified partial ORFs likely to encode two glycosyltransferases, three P-450 monooxygenases and one ABC transporter. The corresponding genes have been found in other glycopeptide gene clusters. Furthermore, upstream to the peptide synthetase region a segment was identified with a remarkable similarity to the vanHAX operon, conferring resistance to glycopeptides in enterococci. Thus, in contrast to the other glycopeptide producers thus far analyzed, in A. teichomyceticus the genes for teicoplanin biosynthesis are closely linked to homologs of glycopeptide resistance commonly found in vancomycin-resistant enterococci. PMID- 11386361 TI - A cluster of genes for the biosynthesis of spinosyns, novel macrolide insect control agents produced by Saccharopolyspora spinosa. AB - Spinosyns A and D are the active ingredients in a family of insect control agents produced by fermentation of Saccharopolyspora spinosa. Spinosyns are 21-carbon tetracyclic lactones to which are attached two deoxysugars. Most of the genes involved in spinosyn biosynthesis are clustered in an 74 kb region of the S. spinosa genome. This region has been characterized by DNA sequence analysis and by targeted gene disruptions. The spinosyn biosynthetic gene cluster contains five large genes encoding a type I polyketide synthase, and 14 genes involved in modification of the macrolactone, or in the synthesis, modification and attachment of the deoxysugars. Four genes required for rhamnose biosynthesis (two of which are also required for forosamine biosynthesis) are not present in the cluster. A pathway for the biosynthesis of spinosyns is proposed. PMID- 11386362 TI - Engineering biodiversity with type II polyketide synthase genes. AB - A very important task in the ongoing search for new clinically useful drugs is the generation of large numbers of structurally diverse compounds. The emerging field of combinatorial biosynthesis, in which nature's chemical capabilities are exploited in a combinatorial 'mix-and-match' fashion, has generated libraries of novel molecules representing great structural diversity which are not available naturally or readily generated through (combinatorial) synthesis. Novel polyketides have been generated by manipulating type II iterative polyketide synthase (PKS) systems that express a variety of combinations of a minimal PKS with ketoreductases, cyclases, and other tailoring enzymes, resulting in a set of design rules to rationally engineer new metabolites. Engineering studies with the Streptomyces coelicolor whiE (spore pigment) and the 'Streptomyces maritimus' enterocin type II PKS provide additional insight on designing diverse assemblies of aromatic, as well as nonaromatic, polyketides. PMID- 11386363 TI - Rare genera of actinomycetes as potential producers of new antibiotics. AB - A literature survey covering more than twenty-three thousand bioactive microbial products including eight thousand antiinfectives demonstrated the increasing relevance of the so called 'rare' actinomycetes as a source of new antibiotics. Past and present efforts in the isolation of rare actinomycetes have enriched the Biosearch Italia Strain Collection with more than twenty thousand strains, showing that, when selective isolation methods are developed and extensively applied, some genera, such as Actinomadura, Actinoplanes, Micromonospora, Microtetraspora, are not rare at all and can be recovered from many soil samples. The current focus is on the isolation of members of Streptosporangiaceae family, given their promising chemical diversity. PMID- 11386364 TI - Agrobacterium rhizogenes-transformed roots of Medicago truncatula for the study of nitrogen-fixing and endomycorrhizal symbiotic associations. AB - Medicago truncatula, a diploid autogamous legume, is currently being developed as a model plant for the study of root endosymbiotic associations, including nodulation and mycorrhizal colonization. An important requirement for such a plant is the possibility of rapidly introducing and analyzing chimeric gene constructs in root tissues. For this reason, we developed and optimized a convenient protocol for Agrobacterium rhizogenes-mediated transformation of M. truncatula. This unusual protocol, which involves the inoculation of sectioned seedling radicles, results in rapid and efficient hairy root organogenesis and the subsequent development of vigorous "composite plants." In addition, we found that kanamycin can be used to select for the cotransformation of hairy roots directly with gene constructs of interest. M. truncatula composite plant hairy roots have a similar morphology to normal roots and can be nodulated successfully by their nitrogen-fixing symbiotic partner, Sinorhizobium meliloti. Furthermore, spatiotemporal expression of the Nod factor-responsive reporter pMtENOD11-gusA in hairy root epidermal tissues is indistinguishable from that observed in Agrobacterium tumefaciens-transformed lines. M. truncatula hairy root explants can be propagated in vitro, and we demonstrate that these clonal lines can be colonized by endomycorrhizal fungi such as Glomus intraradices with the formation of arbuscules within cortical cells. Our results suggest that M. truncatula hairy roots represent a particularly attractive system with which to study endosymbiotic associations in transgenically modified roots. PMID- 11386365 TI - Transposon mutagenesis of Xylella fastidiosa by electroporation of Tn5 synaptic complexes. AB - Pierce's disease, a lethal disease of grapevine, is caused by Xylella fastidiosa, a gram-negative, xylem-limited bacterium that is transmitted from plant to plant by xylem-feeding insects. Strains of X. fastidiosa also have been associated with diseases that cause tremendous losses in many other economically important plants, including citrus. Although the complete genome sequence of X. fastidiosa has recently been determined, the inability to transform or produce transposon mutants of X. fastidiosa has been a major impediment to understanding pathogen-, plant-, and insect-vector interactions. We evaluated the ability of four different suicide vectors carrying either Tn5 or Tn10 transposons as well as a preformed Tn5 transposase-transposon synaptic complex (transposome) to transpose X. fastidiosa. The four suicide vectors failed to produce any detectable transposition events. Electroporation of transposomes, however, yielded 6 x 10(3) and 4 x 10(3) Tn5 mutants per microg of DNA in two different grapevine strains of X. fastidiosa. Molecular analysis showed that the transposition insertions were single, independent, stable events. Sequence analysis of the Tn5 insertion sites indicated that the transpositions occur randomly in the X. fastidiosa genome. Transposome-mediated mutagenesis should facilitate the identification of X. fastidiosa genes that mediate plant pathogenicity and insect transmission. PMID- 11386366 TI - Mutations in pea seedborne mosaic virus genome-linked protein VPg after pathotype specific virulence in Pisum sativum. AB - Pisum sativum plant introduction (PI) line 269818 is resistant to potyvirus pea seedborne mosaic virus (PSbMV) isolates, categorized as pathotype P1, and is susceptible to pathotype P4 isolates. This difference in infectivity is determined by the viral genome-linked protein (VPg) cistron. Mutational analysis of VPg of PSbMV isolates DPD1 and NY representing pathotypes P1 and P4 revealed that codon changes affecting amino acids 105 to 117 in the central region of VPg influenced virulence on PI 269818. In contrast, infectivity on pea cultivar Dark Skinned Perfection, which is susceptible to both pathotypes, was not affected by the mutations. Mutants overcoming resistance in PI 269818 were analyzed for changes in the VPg coding region upon passage through PI 269818 and Dark Skinned Perfection. Adaptive changes were observed only upon passage through PI 269818 and only at codons from amino acid 105 to 117. Expression of DPD1 VPg in PI 269818 did not affect infection by NY, which suggests that VPg from DPD1 is not an elicitor of a general resistance response. The results are compatible with the hypothesis that viral amplification depends upon the interaction between VPg and a host factor. PMID- 11386367 TI - The suppressor of transgene RNA silencing encoded by Cucumber mosaic virus interferes with salicylic acid-mediated virus resistance. AB - The Cucumber mosaic virus (CMV)-encoded 2b protein (Cmv2b) is a nuclear protein that suppresses transgene RNA silencing in Nicotiana benthamiana. Cmv2b is an important virulence determinant but nonessential for systemic spread in N. glutinosa, in contrast to its indispensable role for systemic infections in cucumber. Here, we report that Cmv2b became essential for systemic infections in older N. glutinosa plants or in young seedlings pretreated with salicylic acid (SA). Expression of Cmv2b from the genome of either CMV or Tobacco mosaic virus significantly reduced the inhibitory effect of SA on virus accumulation in inoculated leaves and systemic leaves. A close correlation is demonstrated between Cmv2b expression and a reduced SA-dependent induction of the alternative oxidase gene, a component of the recently proposed SA-regulated antiviral defense. These results collectively reveal a novel activity of Cmv2b in the inhibition of SA-mediated virus resistance. We used a N. tabacum line expressing a bacterial nahG transgene that degrades SA to provide evidence for a Cmv2b sensitive antiviral defense mechanism in tobacco in which SA acts as a positive modifier but not as an essential component. We propose that SA induces virus resistance by potentiating a RNA-silencing antiviral defense that is targeted by Cmv2b. PMID- 11386368 TI - Induction of plant gp91 phox homolog by fungal cell wall, arachidonic acid, and salicylic acid in potato. AB - The oxidative burst has been suggested to be a primary event responsible for triggering the cascade of defense responses in various plant species against infection with avirulent pathogens or pathogen-derived elicitors. The molecular mechanisms of rapid production of active oxygen species (AOS), however, are not well known. We isolated homologs of gp91 phox, a plasma membrane protein of the neutrophil NADPH oxidase, from a potato cDNA library. Molecular cloning of the cDNA showed that there are two isogenes, designated StrbohA and StrbohB, respectively. The RNA gel blot analyses showed that StrbohA was constitutively expressed at a low level, whereas StrbohB was induced by hyphal wall components (HWC elicitor) from Phytophthora infestans in potato tubers. Treatment of potato tubers with HWC elicitor caused a rapid but weak transient accumulation of H2O2 (phase I), followed by a massive oxidative burst 6 to 9 h after treatment (phase II). Diphenylene iodonium (DPI), an inhibitor of the neutrophil NADPH oxidase, blocked both bursts, whereas pretreatment of the protein synthesis inhibitor cycloheximide with the tuber abolished only the second burst. These results suggest that the expression of StrbohA and StrbohB contributes to phase I and II bursts, respectively. The same is true for arachidonic acid, a lipid component of P. infestans-stimulated biphasic oxidative burst, whereas an endogenous signaling molecule, salicylic acid, only induced a weak phase II burst. Both molecules induced the StrbohB expression, which is in agreement with the second burst. To characterize the signal transduction pathway leading to the oxidative burst, we examined the role of protein phosphorylation in HWC-stimulated StrbohB gene expression. K252a and staurosporine, two protein kinase inhibitors, blocked the transcript accumulation. Two inhibitors of extracellular Ca2+ movement, however, did not abolish the transcript accumulation of StrbohB, suggesting that certain calcium-independent protein kinases are involved in the process of StrbohB gene expression. Additionally, we examined a causal relationship between the oxidative burst and expression of defense genes induced by the HWC elicitor. The transcript accumulation of genes related to sesquiterpenoid phytoalexin synthesis (lubimin and rishitin) and phenylpropanoid pathway was inhibited slightly by the DPI treatment, suggesting that the oxidative burst is not essential to activate these genes. Interestingly, the concomitant presence of DPI with the elicitor resulted in an increase in lubimin accumulation and a decrease in rishitin accumulation. Because it is known that lubimin is metabolized into rishitin via oxylubimin, we propose that AOS mediates the synthesis of rishitin from lubimin. PMID- 11386369 TI - Medicago truncatula ENOD11: a novel RPRP-encoding early nodulin gene expressed during mycorrhization in arbuscule-containing cells. AB - Leguminous plants establish endosymbiotic associations with both rhizobia (nitrogen fixation) and arbuscular mycorrhizal fungi (phosphate uptake). These associations involve controlled entry of the soil microsymbiont into the root and the coordinated differentiation of the respective partners to generate the appropriate exchange interfaces. As part of a study to evaluate analogies at the molecular level between these two plant-microbe interactions, we focused on genes from Medicago truncatula encoding putative cell wall repetitive proline-rich proteins (RPRPs) expressed during the early stages of root nodulation. Here we report that a novel RPRP-encoding gene, MtENOD11, is transcribed during preinfection and infection stages of nodulation in root and nodule tissues. By means of reverse transcription-polymerase chain reaction and a promoter-reporter gene strategy, we demonstrate that this gene is also expressed during root colonization by endomycorrhizal fungi in inner cortical cells containing recently formed arbuscules. In contrast, no activation of MtENOD11 is observed during root colonization by a nonsymbiotic, biotrophic Rhizoctonia fungal species. Analysis of transgenic Medicago spp. plants expressing pMtENOD11-gusA also revealed that this gene is transcribed in a variety of nonsymbiotic specialized cell types in the root, shoot, and developing seed, either sharing high secretion/metabolite exchange activity or subject to regulated modifications in cell shape. The potential role of early nodulins with atypical RPRP structures such as ENOD11 and ENOD12 in symbiotic and nonsymbiotic cellular contexts is discussed. PMID- 11386370 TI - Endopolygalacturonase is essential for citrus black rot caused by Alternaria citri but not brown spot caused by Alternaria alternata. AB - Alternaria citri, the cause of Alternaria black rot, and Alternaria alternata rough lemon pathotype, the cause of Alternaria brown spot, are morphologically indistinguishable pathogens of citrus: one causes rot by macerating tissues and the other causes necrotic spots by producing a host-selective toxin. To evaluate the role of endopolygalacturonase (endoPG) in pathogenicity of these two Alternaria spp. pathogens, their genes for endoPG were mutated by gene targeting. The endoPGs produced by these fungi have similar biochemical properties, and the genes are highly similar (99.6% nucleotide identity). The phenotypes of the mutants, however, are completely different. An endoPG mutant of A. citri was significantly reduced in its ability to cause black rot symptoms on citrus as well as in the maceration of potato tissue and could not colonize citrus peel segments. In contrast, an endoPG mutant of A. alternata was unchanged in pathogenicity. The results indicate that a cell wall-degrading enzyme can play different roles in the pathogenicity of fungal pathogens. The role of a cell wall degrading enzyme depends upon the type of disease but not the taxonomy of the fungus. PMID- 11386371 TI - Essential role of superoxide dismutase on the pathogenicity of Erwinia chrysanthemi strain 3937. AB - The sodA gene from Erwinia chrysanthemi strain 3937 was cloned by functional complementation of an Escherichia coli sodA sodB mutant and sequenced. We identified a 639-bp open reading frame, which encodes a protein that is 85% identical to the E. coli manganese-containing superoxide dismutase MnSOD. Promoter elements of this gene were identified by transcriptional mapping experiments. We constructed an E. chrysanthemi deltasodA mutant by reverse genetics. The deltasodA mutation resulted in the absence of a cytoplasmic SOD, which displays the same characteristics as those of MnSOD. The deltasodA mutant was more sensitive to paraquat than the wild-type strain. This mutant could macerate potato tubers, similar to the wild-type strain. In contrast, when inoculated on African violets, the mutant produced, at most, only small necrotic lesions. If the inoculum was supplemented with the superoxide anion-scavenging metalloporphyrin MnTMPyP or purified SOD and catalase, the deltasodA mutant was able to macerate the inoculated zone. Generation of superoxide anion by African violet leaves inoculated with E. chrysanthemi was demonstrated with nitroblue tetrazolium as an indicator. Therefore, at the onset of infection, E. chrysanthemi cells encounter an oxidative environment and require active protective systems against oxidative damages such as MnSOD to overcome these types of conditions. PMID- 11386372 TI - Expression of the gum operon directing xanthan biosynthesis in Xanthomonas campestris and its regulation in planta. AB - The gum gene cluster of Xanthomonas campestris pv. campestris comprises 12 genes whose products are involved in the biosynthesis of the extracellular polysaccharide xanthan. These genes are expressed primarily as an operon from a promoter upstream of the first gene, gumB. Although the regulation of xanthan synthesis in vitro has been well studied, nothing is known of its regulation in planta. A reporter plasmid was constructed in which the promoter region of the gum operon was fused to gusA. In liquid cultures, the expression of the gumgusA reporter was correlated closely with the production of xanthan, although a low basal level of beta-glucuronidase activity was seen in the absence of added carbon sources when xanthan production was very low. The expression of the gumgusA fusion also was subject to positive regulation by rpfF, which is responsible for the synthesis of the diffusible signal factor (DSF). The expression of the gumgusA fusion in bacteria recovered from inoculated turnip leaves was maximal at the later phases of growth and was subject to regulation by rpfF. These results provide indirect support for the operation of the DSF regulatory system in bacteria in planta. PMID- 11386373 TI - Root mucilage from pea and its utilization by rhizosphere bacteria as a sole carbon source. AB - Plant roots secrete a complex polysaccharide mucilage that may provide a significant source of carbon for microbes that colonize the rhizosphere. High molecular weight mucilage was separated by high-pressure liquid chromatography gel filtration from low molecular weight components of pea root exudate. Purified pea root mucilage generally was similar in sugar and glycosidic linkage composition to mucilage from cowpea, wheat, rice, and maize, but appeared to contain an unusually high amount of material that was similar to arabinogalactan protein. Purified pea mucilage was used as the sole carbon source for growth of several pea rhizosphere bacteria, including Rhizobium leguminosarum 8401 and 4292, Burkholderia cepacia AMMD, and Pseudomonas fluorescens PRA25. These species grew on mucilage to cell densities of three- to 25-fold higher than controls with no added carbon source, with cell densities of 1 to 15% of those obtained on an equal weight of glucose. Micromolar concentrations of nod gene-inducing flavonoids specifically stimulated mucilage-dependent growth of R. leguminosarum 8401 to levels almost equaling the glucose controls. R. leguminosarum 8401 was able to hydrolyze p-nitrophenyl glycosides of various sugars and partially utilize a number of purified plant polysaccharides as sole carbon sources, indicating that R. leguminosarum 8401 can make an unexpected variety of carbohydrases, in accordance with its ability to extensively utilize pea root mucilage. PMID- 11386374 TI - Induction of hydroxycinnamoyl-tyramine conjugates in pepper by Xanthomonas campestris, a plant defense response activated by hrp gene-dependent and hrp gene independent mechanisms. AB - Inoculation of pepper leaves, Capsicum annuum cv. Early Calwonder ECW 10R, with strains of Xanthomonas campestris led to an accumulation of the phenolic conjugates feruloyltyramine (FT) and p-coumaroyltyramine (CT) 24 h postinoculation in nonhost- and gene-for-gene-determined incompatible interactions with X. campestris pv. campestris and X. campestris pv. vesicatoria, respectively. In contrast, neither compound was detected in compatible interactions with X. campestris pv. vesicatoria. The accumulation of FT and CT was preceded by an increase in the extractable activity of tyrosine decarboxylase as well as increases in the transcription of genes encoding phenylalanine ammonia lyase and tyramine hydroxycinnamoyl transferase. No such changes were detected in compatible interactions. Very rapid accumulation of FT and CT occurred (4 h postinoculation) in pepper in response to a X. campestris pv. campestris mutant carrying a deletion of the hrp gene cluster. In contrast, hrp mutants of X. campestris pv. vesicatoria failed to elicit the production of FT and CT. These observations suggest the existence of hrp gene-dependent and -independent activation mechanisms of a defense response involving hydroxycinnamoyltyramines. PMID- 11386375 TI - Intracellular accumulation of mannopine, an opine produced by crown gall tumors, transiently inhibits growth of Agrobacterium tumefaciens. AB - pYDH208, a cosmid clone from the octopine-mannityl opine-type tumor-inducing (Ti) plasmid pTi15955 confers utilization of mannopine (MOP) and agropine (AGR) on Agrobacterium tumefaciens strain NT1. NT1 harboring pYDH208 with an insertion mutation in mocC, which codes for MOP oxidoreductase, not only fails to utilize MOP as a sole carbon source, but also was inhibited in its growth by MOP and AGR. In contrast, the growth of mutants with insertions in other tested moc genes was not inhibited by either opine. Growth of strains NT1 or UIA5, a derivative of C58 that lacks pAtC58, was not inhibited by MOP, but growth of NT1 or UIA5 harboring pRE10, which codes for the MOP transport system, was inhibited by the opine. When a clone expressing mocC was introduced, the growth of strain NT1(pRE10) was not inhibited by MOP, although UIA5(pRE10) was still weakly inhibited. In strain NT1(pRE10, mocC), santhopine (SOP), produced by the oxidation of MOP by MocC, was further degraded by functions encoded by pAtC58. These results suggest that MOP and, to a lesser extent, SOP are inhibitory when accumulated intracellularly. The growth of NT1(pRE10), as measured by turbidity and viable cell counts, ceased upon the addition of MOP but restarted in a few hours. Regrowth was partly the result of the outgrowth of spontaneous MOP-resistant mutants and partly the adaptation of cells to MOP in the medium. Chrysopine, isochrysopine, and analogs of MOP in which the glutamine residue is substituted with other amino acids were barely taken up by NT1(pRE10) and were not inhibitory to growth of the strain. Sugar analogs of MOP were inhibitory, and those containing sugars in the D form were more inhibitory than those containing sugars in the L form. MOP analogs containing hexose sugars were more inhibitory than those containing sugars with three, four, or five carbon atoms. Mutants of NT1(pRE10) that are resistant to MOP arose in the zone of growth inhibition. Genetic and physiological analyses indicate that the mutations are located on pRE10 and abolish uptake of the opine. PMID- 11386376 TI - Lettuce mosaic virus pathogenicity determinants in susceptible and tolerant lettuce cultivars map to different regions of the viral genome. AB - Full-length infectious cDNA clones were constructed for two isolates (LMV-0 and LMV-E) of Lettuce mosaic virus (LMV), a member of the genus Potyvirus. These two isolates differ in pathogenicity in susceptible and tolerant-resistant lettuce cultivars. In susceptible plants, LMV-0 induces mild mosaic symptoms, whereas LMV E induces severe stunting, leaf deformation, and a necrotic mosaic. In plants carrying either of the two probably allelic recessive resistance genes mol1 or mol2, LMV-0 is restricted partially to the inoculated leaves. When a systemic invasion does occur, however, symptoms fail to develop. LMV-E overcomes the protection afforded by the resistance genes, resulting in systemic mosaic symptoms. Analysis of the behavior of recombinants constructed between the two virus isolates determined that the HC-Pro protein of LMV-E causes the severe stunting and necrotic mosaic induced by this isolate in susceptible cultivars. In contrast, the ability to overcome mol resistance and induce symptoms in the resistant-tolerant cultivars was mapped to the 3' half of the LMV-E genome. These results indicate that the ability to induce severe symptoms and to overcome the protection afforded by the recessive genes mol1 or mol2 are independent phenomena. PMID- 11386377 TI - The disruption of a gene encoding a putative arylesterase impairs pyruvate dehydrogenase complex activity and nitrogen fixation in Sinorhizobium meliloti. AB - Nitrogen-fixing Sinorhizobium meliloti cells depend upon dicarboxylic acids as carbon and energy sources. The metabolism of these intermediate compounds of the trichloroacetic acid cycle is dependent upon the availability of acetyl-coenzyme A (CoA). In bacteroids, the combined activities of malic enzymes and pyruvate dehydrogenase (PDH) have been proposed to be responsible for the anaplerotic synthesis of acetyl-CoA. We obtained a S. meliloti mutant strain, PD3, in which a Tn5 insertion led to a significant decrease in the overall PDH activity. The genetic characterization of this mutant revealed that the transposon is located at the 3' end of a gene (ada) encoding a putative arylesterase. The mutant PD3 is deficient in nitrogen fixation, which strengthens the physiological importance of PDH activity in the symbiosis of S. meliloti with alfalfa plants. PMID- 11386378 TI - Control of exuT activity for galacturonate transport by the negative regulator ExuR in Erwinia chrysanthemi EC16. AB - The negative regulatory protein ExuR in Erwinia chrysanthemi regulates expression of the galacturonate uptake (exuT) and utilization (uxaA, uxaB, uxaC) genes. We cloned and determined the nucleotide sequence of the exuR gene from E. chrysanthemi EC16. Analysis of the deduced amino acid sequence indicates that this protein possesses a helix-turn-helix motif and belongs to the GntR family of transcriptional repressors. Northern blot analysis and studies with transcriptional fusions of exuT in wild-type and exuR mutant backgrounds indicate that exuT transcription is deregulated in the exuR strain in vivo and in planta. [14C]-galacturonic acid uptake was constitutively high under inducing and noninducing conditions in the exuR mutant. Maximal exuT transcription activity was observed within 8 h of bacterial inoculation into potato tubers, well before any visible symptoms of disease were detected. This suggests that ExuT transport activity in E. chrysanthemi is important in the early stages of disease development. PMID- 11386379 TI - Myocardial infarction with normal coronary arteries: the pathologic and clinical perspectives. AB - Myocardial infarction with normal coronary arteries is a syndrome resulting from numerous conditions but the exact cause in a majority of the patients remains unknown. Cigarette smokers and cocaine users are more prone to develop this condition. The possible mechanisms causing myocardial infarction with normal coronary arteries are hypercoagulable states, coronary embolism, an imbalance between oxygen demand and supply, intense sympathetic stimulation, non atherosclerotic coronary diseases, coronary trauma, coronary vasospasm, coronary thrombosis, and endothelial dysfunction. It primarily affects younger individuals, and the clinical presentation is similar to that of myocardial infarction with coronary atherosclerosis. Thrombolytics, aspirin, nitrates, and beta blockers should be instituted as a standard therapy for acute myocardial infarction. Once normal coronary arteries are identified on subsequent angiography, the calcium channel blockers could be added since coronary vasospasm appears to play a major role in the pathophysiology of this condition. The beta blockers should be avoided in cocaine-induced myocardial infarction because the coronary spasm may worsen. In myocardial infarction with normal coronary arteries, complications such as malignant arrhythmia, heart failure, and hypotension are generally less common, and prognosis is usually good. Recurrent infarction, postinfarction angina, heart failure, and sudden cardiac death are rare. Stress electrocardiography and imaging studies are not useful prognostic tests and long-term survival mainly depends on the residual left ventricular function, which is usually good. PMID- 11386380 TI - Safety and feasibility of two-dimensional echocardiography and myocardial perfusion imaging in patients with chest pain. AB - The accurate identification of patients at high risk for acute coronary syndromes among those seen in the emergency department with possible myocardial ischemia and nonischernic electrocardiograms is a problem. Two-dimensional echocardiography and myocardial perfusion imaging with 99m-technetium sestamibi can identify patients at low and high risk; however, comparative studies are lacking. The authors studied 555 patients considered at low or moderate risk for myocardial ischemia in our emergency department on the basis of the presenting history, and results of physical examination and electrocardiography. These patients underwent echocardiography and myocardial perfusion imaging within 4 hours of presentation. Endpoints included myocardial infarction, percutaneous transluminal coronary angioplasty, and positive results on stress perfusion imaging. Both imaging procedures were performed in the emergency department on 370 patients. Overall agreement between the two techniques was high (concordance, 89%) in the patients who had myocardial infarction or underwent coronary angiography. Agreement between the two techniques is high when used in patients with possible myocardial ischemia. Both techniques helped identify patients at high risk who required admission and those who could be safely discharged. PMID- 11386381 TI - Diurnal variation of QT dispersion in patients with and without coronary artery disease. AB - QT dispersion defined as interlead QT variability in a 12-lead electrocardiogram was proposed by Day and associates as a simple method to evaluate the repolarization heterogenicity of the ventricular myocardium. The frequency of onset of myocardial infarction and sudden death has been reported to have a circadian variation, with a peak incidence in the early morning hours. The authors investigated whether there is diurnal variation of QT interval and QT interval dispersion in healthy subjects and in patients with coronary artery disease. The study population consisted of two groups. Group I consisted of 62 subjects without coronary artery disease and group II consisted of 82 patients with coronary artery disease. Twelve-lead ECG was recorded for each patient in the morning (between 7 AM and 8 AM), afternoon (between 3 PM and 5 PM) and at night (between 11 PM and 1 AM), on the day after performance of coronary angiography. QTc dispersion was significantly higher in patients with coronary artery disease than in healthy subjects in the morning hours and afternoon (p<0.001). Although the differences were much prominent in group I than group II, both QTc dispersion of morning and afternoon were significantly greater than those at night. There were no statistically significant differences between group I and group II at nighttime with respect to maximum QTc, minimum QTc intervals, and QTc dispersion (p>0.05). In conclusion, QT dispersion shows diurnal variation with an increase in the morning hours in both patients with coronary artery disease and subjects without coronary artery disease. The mechanism of diurnal variation of QT dispersion in patients with coronary artery disease is quite different from that of healthy subjects. PMID- 11386382 TI - Cardiac troponin I levels in patients with left heart failure and cor pulmonale. AB - Cardiac troponin levels are regarded as the most specific of currently available biochemical markers of myocardial damage. Elevated levels of troponin have been previously reported in patients with left heart failure, reflecting small areas of undetected myocardial cell death. The aim of this study was to compare the levels of the cardiac troponin I (cTnI) in patients with left- and right-sided heart failure. Cardiac troponin I levels were studied with immunochemical methods in patients with right heart failure (n = 17) resulting from chronic obstructive pulmonary disease, ischemic left heart failure (n = 23), and nonischemic left heart failure (n = 18) who were admitted to departments of cardiology and chest diseases. Also, cTnI levels were measured in 32 healthy subjects as control group. Protein markers of myocardial injury (cTnI and myoglobin) in patients with left and right heart failure were collected approximately 12 to 36 hours after onset of obvious symptoms. Serum creatine kinase MB band was determined on admission and thereafter twice a day during the first 3 days. Elevated levels of serum cTnI were found in patients with nonischemic (0.83 +/- 0.6 ng/mL, p<0.01) and ischemic left heart failure (0.9 +/- 0.5 ng/mL, p<0.01) when compared to healthy subjects, whereas serum cTnI levels in patients with right heart failure due to chronic obstructive pulmonary disease were not significantly different from those of control subjects (0.22 +/- 0.1 vs 0.16 +/- 0.1 ng/mL, p>0.05). In addition, creatine kinase MB band and myoglobin levels were not significantly different between patient and healthy groups. The mean of cTnI levels in ischemic and even nonischemic left heart failure were increased compared to the mean of values in healthy individuals but without significant creatine kinase MB band and myoglobin elevations. But cTnI levels were not increased in patients with right heart failure due to chronic obstructive pulmonary disease. These data indicate that the cTnI levels are abnormal in left heart failure but not in cor pulmonale. PMID- 11386383 TI - Percutaneous balloon pericardiotomy for patients with malignant pericardial effusion including three malignant pleural mesotheliomas. AB - Ten patients were enrolled in this study to evaluate the therapeutic value of percutaneous balloon pericardiotomy in patients with symptomatic pericardial effusion secondary to malignant diseases. Four patients had breast cancer; 2 had lung cancer; 1 had non-Hodgkin's lymphoma; and 3 had malignant pleural mesothelioma, which is commonly seen in Central Anatolian region of Turkey. All patients underwent percutaneous balloon pericardiotomy with monofoil balloons (Mansfield, NuMed). No complication was seen during these procedures. In 3 patients, the balloon could not be expanded completely and was entered from a more lateral position by a second puncture. There was no recurrence of pericardial effusion in 6 of 7 patients without mesothelioma. After percutaneous balloon pericardiotomy, surgical subxiphoid windowing was performed due to drainage greater than 100 mL/day in a patient with lung cancer and in 1 patient with mesothelioma. In the other 2 patients with mesothelioma, recurrence of pericardial effusion was seen and then subxiphoid surgical windowing was performed due to development of cardiac tamponade in 1 of them. All the patients died 68.6 +/- 36 days later due to the primary malignancies. The survival time of patients with mesothelioma was shorter than that of the others (p < 0.05). These results suggest that percutaneous balloon pericardiotomy may be used in the treatment of patients with malignant pericardial effusion as an alternative to surgical pericardial window creation. But in patients with malignant pleural mesothelioma, the success rate of this procedure was lower than that of the others. PMID- 11386385 TI - Paget-Schroetter syndrome in sports activities--case study and literature review. AB - The authors report 7 patients with thromboses in the upper extremity resembling Paget-Schroetter syndrome. According to their case histories, all patients had a temporal and causal relationship between partially unusual sports activities and the genesis of the thrombosis. The cause of this condition is a strain on the subclavian and axillary veins by retroversion or hyperabduction of the arm. This can entail microtraumatizations of the venous intima, consequently leading to a consecutive local activation of coagulation and to a possible thrombosis of the vessel. A mechanical compression of the vein by adjoining bone, ligament, and muscle structures can intensify the effects. Further primary diseases and risk factors as secondary causes for thromboses where taken into consideration when examining the patients. The Paget-Schroetter syndrome should be considered as a possible cause for unspecified trouble in the upper extremity reported by athletes. If such prolapses occur, they can be categorized as accidents by private and statutory insurance companies that cover accidents. PMID- 11386384 TI - Usefulness of saturation pulses in magnetic resonance imaging of partial anomalous pulmonary venous return. AB - The authors evaluated partial anomalous pulmonary venous return by magnetic resonance (MR) images. Seven patients with this congenital anomaly underwent MR imaging examination. Conventional spin-echo and gradient-echo imaging were performed. In addition, during acquisition of gradient-echo images, saturation pulses were imposed on the affected lung. Spin-echo images showed the anatomical situation of the anomalous veins, and gradient-echo images revealed the blood flow in the veins. With saturation technique, the direction and drainage of blood flow in the anomalous veins were well defined. The study suggests that MR imaging with spin-echo method and gradient-echo method with or without saturation pulses is a useful and noninvasive method of diagnosing partial anomalous pulmonary venous return. MR images with spin- and gradient-echo methods were useful in defining the anatomical situation and blood flow in the anomalous veins. By imposing saturation pulses on the affected lung field, the direction and drainage of blood flow in the anomalous veins were clearly demonstrated. PMID- 11386386 TI - Dilated cardiomyopathy of Becker-type muscular dystrophy with exon 4 deletion--a case report. AB - The authors report a 47-year-old man with Becker-type muscular dystrophy presenting with dilated cardiomyopathy. Left ventriculography showed diffuse severe hypokinesia: left ventricular end-diastolic volume index 193 mL/m2, left ventricular end-systolic volume index 143 mL/m2, and left ventricular ejection fraction 26%. Skeletal muscle biopsy demonstrated a dystrophic process. Genetic analysis revealed a deletion of exon 4. There was a difference in immunostaining pattern between skeletal muscles and cardiac muscles. Severe cardiac dysfunction in this case may be associated with the damage in dystrophin-deficient fibers. PMID- 11386387 TI - St. Anthony's fire (ergotamine induced leg ischemia)--a case report and review of the literature. AB - Ergotism, once an epidemic disease, is now a rare disorder. The most common manifestation is acute peripheral ischemia due to vasospasm, with an incidence of 0.001%. We report a case of a middle-age woman who presented with ergotamine induced leg ischemia, due to chronic use of ergotamine-containing medications for migraine headaches. The diagnosis was confirmed with arteriography results, and she responded well to vasodilator therapy. The pharmacology, clinical presentation, diagnostic approach, and therapy of ergotism are reviewed. PMID- 11386388 TI - Contrast harmonic power Doppler imaging of congenital ventricular diverticulum--a case report. AB - A case of an 81-year-old woman with a left ventricular diverticulum who underwent myocardial contrast echocardiography is reported. After administration of the contrast agent, a pulsed Doppler flow measurement clearly revealed the biphasic waveform of the ejection flow in the pre-systolic and systolic phase at the ostium of the diverticular cavity. A harmonic power Doppler image showed that part of the diverticulum wall had similar acoustic properties to the ventricular septal wall. Intracardiac blood flow and myocardial perfusion could be clearly evaluated and a ventricular diverticulum was correctly diagnosed using contrast echocardiography. PMID- 11386389 TI - Rapid clot formation and abnormal fibrin structure in a symptomatic patient taking fenfluramine--a case report. AB - A 35-year-old woman experienced symptomatic calf pain while taking a combination of fenfluramine and phentermine. All symptoms resolved when the medications were stopped, but pain returned when fenfluramine was restarted. Laboratory evaluation revealed mild elevations of aspartate aminotransferase and lactate dehydrogenase and a remarkably shortened prothrombin time (6.3 seconds). Additional studies revealed that the clots were composed of very thin fibrin fibers. All laboratory abnormalities, including the abnormal fibrin structure, completely resolved when fenfluramin was stopped. Direct addition of fenfluramine or phentermine to normal plasma did not alter either coagulation kinetics or fibrin structure, supporting the concept that the induced changes may have originated at the hepatic level. Clots composed of thin fibers are much more resistant to fibrinolysis, and could potentially put such patients at risk for thrombotic complications. This is the first report of clotting abnormalities associated with fenfluramine use. Subsequent to its use in this patient, fenfluramine was removed from clinical use due to reports of acquired valvular heart disease. PMID- 11386390 TI - Revised indications for statin therapies. PMID- 11386391 TI - Research into rehabilitation. What is the priority? PMID- 11386392 TI - A randomized efficacy and feasibility study of imagery in acute stroke. AB - OBJECTIVE: To compare the feasibility and efficacy of a programme combining imagery and occupational therapy with a programme of therapy only. DESIGN: Randomized, controlled case series. SETTING: Subacute outpatient clinic. SUBJECTS: Thirteen consecutively admitted patients between four weeks and one year post stroke exhibiting stable motor deficits in their affected upper limbs. INTERVENTION: All patients received an hour of therapy three times a week for six weeks administered by the same physical and occupational therapists. During the same period, eight patients participated in 10-minute guided imagery sessions after each therapy session, as well as practising imagery at home twice each week. Five patients participated in a control intervention consisting of exposure to stroke information. MAIN OUTCOME MEASURES: The Fugl-Meyer Assessment of Motor Recovery (Fugl-Meyer) and Action Research Arm Test (ARA). RESULTS: After intervention, Fugl-Meyer and ARA scores of patients in the therapy only group remained virtually the same; therapy plus imagery group scores improved by 13.8 and 16.4 points, respectively, on the Fugl-Meyer and ARA. CONCLUSIONS: Imagery is a clinically feasible, cost-effective complement to therapy that may improve outcomes more than participation in therapy only. PMID- 11386393 TI - The Parkinson's Disease Activities of Daily Living Scale: a new simple and brief subjective measure of disability in Parkinson's disease. AB - OBJECTIVE: To develop a brief, valid and reliable self-report scale for the assessment of activities of daily living in Parkinson's disease (PD). DESIGN: Self-report questionnaire development. SUBJECTS: One hundred and seventy subjects with a diagnosis of clinically probable PD living in the community. MEASURES: The self-rating scale--Parkinson's Disease Activities of Daily Living Scale (PADLS), Webster Scale, CAMCOG neuropsychological test,15-item Geriatric Depression Scale (GDS-15) and the self-rated Parkinson's Disease Quality of Life (PDQL) questionnaire. METHODS: The PADLS was initially validated and test-retest reliability assessed in a group of PD patients (n = 38). Next a convenience sample of 132 patients was drawn from a community-based PD register. Subjects were invited to complete the PADLS, PDQL, GDS-15, Webster scale and CAMCOG test. RESULTS: The PADLS correlated significantly with increasing age, duration of illness, disease severity, increasing depression, impaired cognition and poorer health-related quality of life. CONCLUSION: The PADLS was found to be a reliable and valid measure of ADL, demonstrating acceptable internal consistency and strong associations with existing measurers of disease severity, depression, cognitive screening and health-related quality of life. The PADLS allows patients to subjectively report the impact that PD has upon daily activities and will complement existing formal clinical measures in PD. PMID- 11386394 TI - Developing a disease-specific quality of life measure for people with multiple sclerosis. AB - OBJECTIVES: To develop a patient-completed disease-specific measure of quality of life in multiple sclerosis and to validate the measure in a community-based population of people with multiple sclerosis. METHODS: The items in the scale were selected in focus group sessions of people with multiple sclerosis. The initial scale included 25 items and was tested in subgroups of 150 people from a population register of people with multiple sclerosis in Leeds. Following further developmental phases, a restructured 16-item scale was tested on a random sample of 200 people with multiple sclerosis from the population register, stratified according to disease course. This led to a final eight-item unidimensional scale, the Leeds Multiple Sclerosis Quality of Life (LMSQoL) scale. RESULTS: After initial development a 16-item scale was found to be both reliable and valid. Cronbach's alpha for the 16-item scale was 0.86. The test-retest correlation was 0.74, using a two-week retest interval. However, convergent validity with the General Well Being Index was 0.67 and with the SF-36 Physical Function Scale was 0.68. This suggested that the scale straddled these two concepts and was confirmed by fit of the data to the Rasch measurement model. This revealed the potential for a reduced eight-item version of the scale. The eight-item scale had a closer association to well-being (0.83) than to physical function (0.39), had good internal consistency (0.79) and test-retest reliability (0.85). There were virtually no floor or ceiling effects for the scale. CONCLUSIONS: The study presents a disease-specific measure of quality of life in multiple sclerosis, the Leeds Multiple Sclerosis Quality of Life (LMSQoL) scale. The instrument is brief, easy to use and practical to administer in clinic or as a postal questionnaire. It measures a construct related to well-being, and provides an important adjunct to the measurement of outcome in multiple sclerosis. PMID- 11386395 TI - Measuring the impact of multiple sclerosis on psychosocial functioning: the development of a new self-efficacy scale. AB - OBJECTIVE: To develop a scale to measure self-efficacy in neurologically impaired patients with multiple sclerosis and to assess the scale's psychometric properties. DESIGN: Cross-sectional questionnaire study in a clinical setting, the retest questionnaire returned by mail after completion at home. SETTING: Regional multiple sclerosis (MS) outpatient clinic or the Clinical Trials Unit (CTU) at a large neuroscience centre in the UK. SUBJECTS: One hundred persons with MS attending the Walton Centre for Neurology and Neurosurgery and Clatterbridge Hospital, Wirral, as outpatients. METHODS: Cognitively impaired patients were excluded at an initial clinic assessment. Patients were asked to provide demographic data and complete the self-efficacy scale along with the following validated scales: Hospital Anxiety and Depression Scale, Rosenberg Self Esteem Scale, Impact, Stigma and Mastery and Rankin Scales. The Rankin Scale and Barthel Index were also assessed by the physician. RESULTS: A new 11-item self efficacy scale was constructed consisting of two domains of control and personal agency. The validity of the scale was confirmed using Cronbach's alpha analysis of internal consistency (alpha = 0.81). The test-retest reliability of the scale over two weeks was acceptable with an intraclass correlation coefficient of 0.79. Construct validity was investigated using Pearson's product moment correlation coefficient resulting in significant correlations with depression (r= -0.52) anxiety (r =-0.50) and mastery (r= 0.73). Multiple regression analysis demonstrated that these factors accounted for 70% of the variance of scores on the self-efficacy scale, with scores on mastery, anxiety and perceived disability being independently significant. CONCLUSION: Assessment of the psychometric properties of this new self-efficacy scale suggest that it possesses good validity and reliability in patients with multiple sclerosis. PMID- 11386396 TI - Base rate of post-concussion symptoms among normal people and its neuropsychological correlates. AB - OBJECTIVE: To explore the base rate of symptoms similar to those of post concussion symptoms (PCS) among a group of participants without head injury. The effect of subjective complaints upon cognitive functioning was also examined. DESIGN: A cross-sectional study. PARTICIPANTS: A total of 85 participants without head injury, other identifiable neurological diseases or psychiatric diseases were recruited. OUTCOME MEASURES: These included Rivermead Post-concussion Symptoms Checklist, Cognitive Failures Questionnaire, Colour Trails Test, Stroop Word-Colour Test, Paced Auditory Serial Addition Test, Symbol Digits Modality Test, Word Fluency Test, Design Fluency Test, Digits Forward and Backward Span Test, and Modified Six Elements Test. RESULTS: A relatively high proportion of the participants reported symptoms similar to those of patients with PCS. These included longer time to think (65.9%), forgetfulness (58.9%), poor concentration (58.9%), fatigue easily (53.5%), and sleep disturbances (50.6%). Gender effect was not significant for either the individual item or the total score of the Rivermead Post-concussion Symptoms Checklist. Moreover, there was no difference found between low symptom reporters and high symptom reporters in terms of attention, working memory, mental fluency, and strategy allocation. CONCLUSION: The base rates of symptoms were consistent with previous studies among the nonclinical groups. However, persons reporting a high score of concussion-like symptoms did not perform less well than those reporting a low score of symptoms in attention, working memory, mental fluency and strategy allocation. PMID- 11386398 TI - Randomized controlled trial of quadriceps training after proximal femoral fracture. AB - OBJECTIVE: To determine whether systematic progressive high-intensity quadriceps training increases leg extensor power and reduces disability in patients rehabilitating after proximal femoral fracture. DESIGN: Open parallel group randomized controlled trial comparing the addition of six weeks quadriceps training (40 patients) with standard physiotherapy alone (40 patients). The training group exercised twice weekly, with six sets of 12 repetitions of knee extension (both legs), progressing up to 80% of their one-repetition maximum. SETTING: Orthogeriatric unit, and community follow-up. SUBJECTS: Eighty patients rehabilitating after proximal femoral fracture. MAIN OUTCOME MEASURES: Measurements of leg extensor power (Nottingham Power Rig), functional mobility (elderly mobility score), disability (Barthel Index) and quality of life (Nottingham Health Profile) were made at baseline, after six weeks (at the end of the intervention) and at 16 weeks. RESULTS: Leg extensor power increased significantly in the quadriceps training group (fractured leg mean improvement at six weeks 157% (standard error 16), nonfractured leg 80% (12)) compared with the control group (63% (11) and 26% (8) respectively, unpaired Student's t-test p = 0.007 and p = 0.01 for between-group comparisons). Significant benefits were maintained at 16 weeks. Quadriceps training resulted in a greater increase in elderly mobility scale score compared with standard rehabilitation (between-group difference of 2.5 (95% CI 1.1,3.8) at week 6 and 1.9 (0.4,3.4) at week 16). Barthel score increased significantly from week 0 to 6 in the quadriceps training group compared with controls (Mann-Whitney U-test p = 0.05). Patients in the quadriceps training group scored significantly better in the energy subscore of the Nottingham Health Profile at the end of follow-up (Mann-Whitney U-test p = 0.0185). CONCLUSIONS: Progressive high-intensity quadriceps training in elderly proximal femoral fracture patients increased leg extensor power and reduced disability. This was accompanied by an increase in energy as measured by the Nottingham Health Profile. This intervention may provide a simple practical way of improving outcome in these patients. PMID- 11386397 TI - Quadriceps muscle weakness following acute hemiplegic stroke. AB - OBJECTIVE: To determine whether quadriceps muscle weakness develops on the side ipsilateral to the brain lesion in the first week following acute ischaemic hemiplegic stroke. DESIGN: A prospective study of muscle strength. SETTING: Acute stroke unit (ASU) in a teaching hospital. SUBJECTS: Ten patients admitted within 48 hours of stroke onset, and 10 healthy age-matched controls. INTERVENTIONS: Repeat nonvolitional measurements of quadriceps muscle strength of the unaffected limb in patients and the right leg in normal subjects using magnetic femoral nerve stimulation (MS), prospectively one week apart. In addition the level of voluntary activation was assessed during a maximum voluntary contraction (MVC) manoeuvre. The Trunk Control Test (TCT) was measured in the patients. RESULTS: The median (95% confidence interval, (CI)) baseline quadriceps twitch tension (Tw Q) and MVC in the control group were 9.4 kg (6.1-12.5 kg) and 37.2 kg (23.8-54.6 kg), and in the stroke group were 7.6 kg (4.4-9.9 kg) and 12.15 kg (7.9-30.8 kg). The median (95% CI) change in Tw Q and MVC respectively between baseline and one week later were 1.75% (-9.8 to 8%) and 5.45% (-15.1 to 22.7%) (NS) in the control group and -16.2% (-6 to -25.9%) and -30.45% (0 to -78.6%) (p < 0.01) in the stroke patients. There was a significant correlation between the percentage fall in Tw Q and both change in TCT (rs = 0.83, p < 0.01) and percentage change in body weight (rs = 0.83, p < 0.01). CONCLUSION: In the first week after acute hemiplegic stroke, weakness develops in the unaffected leg. PMID- 11386399 TI - Too many pre-discharge home assessment visits for older patients? A postal questionnaire survey. AB - OBJECTIVE: To investigate current practice of pre-discharge home assessment visits for older patients. DESIGN: Postal questionnaire survey. SETTING: NHS Trust hospitals in the United Kingdom. SUBJECTS: Occupational therapy departments in 265 NHS Trust hospitals which admit acutely ill older patients. MAIN OUTCOME MEASURES: The number of pre-discharge home visits done, who went, and therapy time spent on home visits. RESULTS: Of 265 NHS Trusts contacted, 239 (90%) replied. Of 238 units, 155 (65%) do between 11 and 40 visits per month, with 25 (11%) doing more than 60. The equivalent of one day per week or more is spent doing home visits by Senior I occupational therapists in 107 (45%) units and by Senior II staff in 126 (53%) units. Carers or relatives, unqualified occupational therapists, social workers and home care managers accompany the patient and occupational therapist on most home visits. CONCLUSION: Therapy perception is that pre-discharge home assessment visits are increasing in number, complexity and involvement of professional time despite little evidence for their effectiveness. Controlled trials are required to assess which patients are likely to benefit. PMID- 11386400 TI - A comparison of mobility assessments in a geriatric day hospital. AB - OBJECTIVE: To assess the sensitivity of the Elderly Mobility Scale (EMS) to detect improvements in mobility after physiotherapy in comparison with the Barthel Index (BI) and Functional Ambulation Category (FAC) in routine clinical day hospital practice. SUBJECTS: Eighty-three patients who completed a course of physiotherapy were studied. METHODS: Each was assessed by an independent physiotherapist before and after a programme of physiotherapy, using the EMS, BI and FAC. RESULTS: The mean age was 79 years (SD 7.7). Fifty-three out of 83 (64%) patients were female and 92% were community dwelling. The median number of physiotherapy sessions undergone by each patient was 9 (range 3-51). All three scales detected an improvement in mobility with physiotherapy (p < 0.001). However, using the EMS, 68 out of 82 (83%) patients had a detectable improvement in mobility compared with only 34 out of 80 (42%) using the BI and 28 out of 81 (35%) using the FAC. Using a matched-pairs comparison, the EMS was significantly more likely to detect an improvement in mobility following physiotherapy within the study group than the BI (p < 0.001) or the FAC (p < 0.001). CONCLUSION: Although all three scales detected improvements in mobility, the EMS detected mobility improvements in a significantly greater number of patients. PMID- 11386401 TI - Reliability of an interview approach to the Functional Independence Measure. AB - OBJECTIVE: To establish the reliability of an interview approach to the Functional Independence Measure (FIM). DESIGN: Two raters were present at the same time during the interviews in the home and did independent ratings of the 18 FIM items. The interview procedure was repeated within a week by another two raters in the clinic. SUBJECTS: Sixty-three stroke survivors (median age 63 years, range 18-71 years) were studied approximately two years after onset. RATERS: Three occupational therapists and one nurse. METHODS: Reliability was calculated as unweighted kappa statistics, percentage agreement (PA), and intraclass correlation coefficient (ICC). RESULTS: Best agreement was found in the motor items of FIM. The kappa statistics showed good to excellent inter-rater values during the same interview except for the Social interaction item. The ICCs based on sum-score for motor (0.92) and social-cognitive items (0.75) respectively, were similar to those reported in the literature. The repeated interview by different raters showed less stable agreement according to kappa values for the items dealing with transfers, locomotion and social-cognition. CONCLUSIONS: FIM assessments showed high inter-rater agreement for the same interview setting (home as well as clinic), but the stability of the measure over time with a repeated interview by different raters was somewhat less satisfactory. PMID- 11386402 TI - Adaptation of the Functional Independence Measure for use in Turkey. AB - OBJECTIVE: To adapt the Functional Independence Measure (FIM) for use in Turkey and to assess its validity and reliability. DESIGN: After the translation procedure, reliability was assessed using internal consistency, inter-rater reliability (kappa) and the intraclass correlation coefficient (ICC). Construct validity was tested by association with impairments and by fit of data to the Rasch model. SETTING: The study was undertaken in an inpatient rehabilitation unit of the Department of Physical Medicine and Rehabilitation, Faculty of Medicine, University of Ankara. SUBJECTS: Consecutive stroke (n = 51) and spinal cord injury (SCI) (n = 62) patients admitted for rehabilitation over a period of three years were assessed at admission and discharge. RESULTS: Internal consistency was good for stroke, and for SCI. The level of agreement between two raters was sufficient with kappa levels of above 0.48 for SCI and above 0.44 for stroke. Minimum ICC found was 0.90. Construct validity showed expected associations with the impairment scales. However, Rasch analysis showed that bladder and bowel items compromise unidimensionality in the motor scale. CONCLUSION: Adaptation of the FIM has been successful and it can be used in Turkey as long as the limitations are recognized. PMID- 11386403 TI - An audit of the provision of environmental control systems in Northern Ireland, 1992-1997. AB - OBJECTIVE: To evaluate the provision of environmental control systems (ECS) in Northern Ireland with regard to assessment and prescription, installation and review and propose guidelines for future service delivery. DESIGN: Structured interview, physical examination, Barthel ADL Index, demonstration and assessment of suitability of ECS for patient. SUBJECTS: Prescriptions for ECS from April 1992 to 1997 were identified from centrally held records. Current users were assessed in their own homes. RESULTS: Forty-six out of 49 current users identified were assessed. All were severely disabled (Barthel 0-9); 24% were living alone; 7 (15%) were not utilizing the system; 96% were satisfied with their initial assessment. Prior to prescription 52% had information about ECS and 20% had a practical demonstration; 78% felt that this would have been useful; 52% of users were not under routine clinical review; 41% of ECS had been altered since installation. Repairs had taken longer than 7 days in 11% of cases. In 45 cases the ECS was essential and in 43 it was appropriate to the users' needs and abilities. CONCLUSIONS: ECS are a valuable tool for severely disabled persons and are appropriately prescribed in Northern Ireland. A multidisciplinary team should perform assessment and prescription. All patients should have a practical trial of the equipment to assist in prescriptions. Regular review by the team should be performed to identify changes in need and alter systems appropriately. Users who live alone should represent a priority for repairs. PMID- 11386404 TI - Position-matching in the upper limb: professional ballet dancers perform with outstanding accuracy. AB - OBJECTIVES: To investigate the accuracy in position-matching in the upper limb in two groups of subjects who were physically fit and movement aware. DESIGN: A mixed-group design was used. Objective measurement of the accuracy in position matching at the shoulder and elbow in both dominant and nondominant arms consisted of photographic record of the position-matching test, with goniometric measurement. SETTINGS: Physiotherapy department at the Birmingham Royal Ballet and School of Health Science, University of Birmingham. SUBJECTS: Two subject groups: physiotherapy students (n = 10), professional ballet dancers (n = 10). RESULTS: A mixed design analysis of variance found significant differences between the accuracy in position-matching at both the shoulder and elbow joints in the two groups (p < 0.05), with the ballet dancers having greater accuracy then the physiotherapy students. A significant difference in the joint positions tested were demonstrated (p < 0.05) with the positions of abduction at the shoulder and extension of the elbow showing greatest accuracy in matching. There was no significant difference found between the dominant and nondominant upper limb in position-matching. CONCLUSION: Professional ballet dancers demonstrated greater accuracy in position-matching the upper limb, implying that mass and continuing practice can improve a motor sensory skill. PMID- 11386405 TI - Muscle strength, pain and disability in patients with osteoarthritis. AB - OBJECTIVE: Reduced muscle strength is regarded as a risk factor for pain and disability in osteoarthritis (OA). Currently, various indices for muscle strength are used when assessing determinants of pain and disability. The goal of the present study was to evaluate these indices of muscle strength. DESIGN: Isometric muscle strength was measured for 16 muscle actions around the knees and hips in 52 patients with OA of the hip and 70 patients with OA of the knee. Various indices of muscle strength were derived from these measurements, applying five alternative approaches. These approaches ranged from a single overall index to a set of 16 separate indices. The internal consistency of these indices was determined (Cronbach's alpha), and it was determined to what extent they could reveal the association between reduced muscle strength on the one hand and pain and disability on the other hand. RESULTS: Internal consistency was satisfactory for all indices (Cronbach's alpha >0.74). As expected, reduced muscle strength was associated with increased disability, but no clear relationship could be established between muscle weakness and pain. The strength of these associations did not depend on the approach used to derive the indices for muscle strength. CONCLUSIONS: The indices did not show major differences with regard to internal consistency or the extent to which the association with pain and disability could be revealed. For reasons of parsimony, approaches resulting in few indices appear to be most useful. However, muscle strength was found to be significantly reduced around affected joints, compared with muscle strength around unaffected joints. Therefore, the most suitable approach for reducing muscle strength data into indices is one that results in as few indices as possible, but with separate indices for muscle strength around affected and unaffected joints. PMID- 11386406 TI - Time-dependent inhibition of glioblastoma cell proliferation by dexamethasone. AB - Because of the outstanding importance of the glucocorticoid Dexamethasone (DEX) as supportive therapy in the management of brain tumours, the direct effect of DEX on tumour cell proliferation is of particular interest. Previous in vitro studies led to contradictory results. To characterise more precisely the influence of DEX, we investigated the glioblastoma multiforme (GM) cell lines A172, T98G and 86HG39. Cells were treated with DEX concentrations ranging from 5 x 10(-9) to 5 x 10(-5) M from 24 to 240h under different treatment conditions. Influence of DEX on glioma cell viability was assessed daily for 5 days by MTT assay: (I) with continuous DEX incubation (acute treatment), (II) in a recultivation period without DEX after 5 days of DEX pre-incubation (pre treatment), (III) with continuous DEX incubation after 5 days of DEX pre incubation (combination treatment). DEX acute treatment led to strongly decreased proliferation of A172 cells, whereas T98G and 86HG39 cells remained uninfluenced. In opposite, a time-delayed inhibition of cell proliferation was observed in all three cell lines after DEX pre-treatment. Combination treatment induced a significant increase of the inhibitory effect in A172 and T98G cells. These data show a variable, partial time-dependent inhibitory effect of DEX on the proliferation of GM cells and may open new treatment strategies for malignant brain tumours. PMID- 11386407 TI - Overcoming of radioresistance in human gliomas by p21WAF1/CIP1 antisense oligonucleotide. AB - Malignant gliomas are highly resistant tumors against gamma-irradiation and contained overexpression of p21WAF1/CIP1 (p21). Overexpression of p21 enhanced clonogenic survival and suppressed apoptosis after gamma-irradiation in human brain tumor cell lines with or without p53 protein deficiency. The effect of antisense oligonucleotide to p21 against the gamma-irradiation-induced apoptosis and cytotoxicity in malignant glioma cell lines was examined. Antennapedia homeodomain internalization peptide was used as an insertion vector. The high transfection efficiency of Antennapedia homeodomain internalization peptide joined with antisense oligonucleotide was observed. The pretreatment with antisense oligonucleotide enhanced the gamma-irradiation-induced apoptosis and cytotoxicity in radioresistant glioma cells. p21 may represent an important new target for radiosensitization protocols, possibly involving antisense oligonucleotide directed against p21. PMID- 11386408 TI - The relationship between genetic aberrations as detected by comparative genomic hybridization and vascularization in glioblastoma xenografts. AB - Angiogenesis is of vital importance for the growth of solid tumors and constitutes a target for anti-cancer therapy. Glioblastomas (GBMs) are histologically characterized by striking microvascular proliferation. The identification of the mechanism of angiogenesis is of major importance for the further development of anti-angiogenic therapy. Tumor angiogenesis might be the result of a combination of local tissue conditions (especially hypoxia) and specific genetic alterations acquired during oncogenesis. In order to investigate the relationship between genetic aberrations and tumor angiogenesis in GBM xenograft lines, the genetic alterations were examined by Comparative Genomic Hybridization (CGH). Two vascular phenotypes of GBM xenografts could be identified: a well vascularized and a poorly vascularized type. In this model, the poorly vascularized type had a larger number of genetic alterations. However, there was no unequivocal correlation between angiogenesis, growth rate and patterns of genetic alterations as detected by CGH. PMID- 11386409 TI - Primary isolated lymphoma of the fourth ventricle: case report. AB - The authors present the case of a 33-year-old woman presenting with a 4-week history of vertigo and headaches. Physical examination revealed an isolated static cerebellar syndrome. Magnetic resonance imaging showed a homogeneously enhancing tumor located in the fourth ventricle. Complete surgical removal was performed and microscopic examination revealed a high-grade B-cell lymphoma. Postoperative investigations confirmed it to be primary. Only one case of primary isolated lymphoma of the fourth ventricle has been briefly reported in the medical literature. The authors suggest that primary lymphoma must be considered in the list of fourth ventricle tumors. PMID- 11386410 TI - Concurrent accelerated hyperfractionated radiation therapy and carboplatin/etoposide in patients with malignant glioma: long-term results of a phase II study. AB - PURPOSE: Feasibility, antitumor activity and toxicity of accelerated hyperfractionated radiation therapy (Acc Hfx RT) and concurrent carboplatin/etoposide (CBDCA/VP 16) chemotherapy were investigated in patients with malignant glioma. MATERIAL AND METHODS: Seventy-nine patients with either glioblastoma multiforme (GBM) (n = 61) or anaplastic astrocytome (AA) (n = 18) entered into a phase II study on the use of Acc Hfx RT with 60 Gy in 40 fractions in 20 treatment days over 4 weeks and concurrent CBDCA, 200 mg/m2, and VP 16, 200 mg/m2, both given once weekly during the RT course. RESULTS: The median survival time for all 79 patients was 14 months (11 and 44 months for GBM and AA patients, respectively), while the 2- and 4-year survival was respectively 33% and 11% for all patients, 13% and 1.6% for GBM patients, and 100% and 44% for AA patients (p < 0.0001). The median time to progression for all patients was 12 months (9 and 40 months for GBM and AA, respectively), while the 2- and 4-year progression-free survival (PFS) was respectively 28% and 10% (all patients), 10% and 1.7% (GBM) and 89% and 39% (AA) (p < 0.0001). Multivariate analysis showed that age, performance status, and preoperative size of tumor influenced survival in GBM. Only 5 (6%) patients experienced grade 3 leukopenia and 6 (8%) patients experienced grade 3 thrombocytopenia. No late RT-induced toxicity was observed to date. CONCLUSIONS: Although Acc Hfx RT/CBDCA + VP 16 was feasible and little toxic, it failed to improve survival/progression-free survival over that obtained with other currently used regimens. These results do not justify the investigation of this regimen in a phase III trial. PMID- 11386411 TI - Acute hemorrhage in late radiation necrosis of the temporal lobe: report of five cases and review of the literature. AB - Hemorrhage in late cerebral radiation necrosis is a rare complication after radiotherapy for intracranial and extracranial neoplasms. We report 5 cases of acute hemorrhage in late radiation necrosis of the temporal lobe following radiation therapy for nasopharyngeal carcinoma. In a review of the literature, the authors identified a total of 27 such cases. The interval period between the onset of hemorrhage and cranial irradiation is long (mean = 7.8 years). The most prominent histological feature was the proliferation of large, dilated and thin walled new blood vessels in a background of gliosis and fibrinoid necrosis of vessels. Rupture of these thin-walled new blood vessels is the proposed mechanism of hemorrhage in this condition. PMID- 11386412 TI - Superselective intra-arterial carboplatin for treatment of intracranial neoplasms: experience in 100 procedures. AB - BACKGROUND: The results of animal studies suggest that superselective intra arterial infusion allows the permeation of a high concentration of chemotherapeutic agents within intracranial neoplasms. In the present report, we review our clinical experience with the 100 intra-arterial infusions of carboplatin in intracranial neoplasms not responsive to other treatment modalities. METHODS: Carboplatin was infused in 100 separate sessions (24 patients) as a mean dose of 286+/-60 mg/m2 (range 34-377 mg/m2). RMP-7, a bradykinin analog, was used as an adjunct in 28 sessions (6 patients). The infusions were performed through superselective microcatheterization of the following arteries: internal carotid (n = 39), middle cerebral (n = 61), posterior cerebral (n = 21) and anterior cerebral (n = 10). The frequency of neurological and non-neurological complications, and survival were recorded. In a subset of 10 patients, tumor volume was measured by serial magnetic resonance images to assess therapeutic response to therapy. RESULTS: The mean age of the patients was 44.5 years (range 26-67 years); 13 were men. The tumors were classified as glioblastoma multiforme (n = 12), metastatic tumor (n = 1), high grade astrocytoma (n = 6), and anaplastic mixed glioma (n = 5). Follow-up was available for 23 patients (mean 22 months, range 2-69 months). Survival beyond 1 year after initiation of intra-arterial carboplatin therapy was documented in 12 of the 23 patients. A total of 13 neurological complications including seizures (n = 7), transient neurological deficits (n = 5), and ischemic stroke (n = 1) were observed in 100 procedures. A lower frequency of complications occurred in men and in patients who received adjunctive RMP-7. Volumetric analysis of serial magnetic resonance images demonstrated tumor mass reduction in 3 out of 10 patients. An increase in tumor mass ranging from 23% to 230% was observed in the other 7 patients over a period ranging from 2.3 to 37.7 months since initiation of carboplatin therapy. CONCLUSIONS: Superselective intra-arterial administration of carboplatin appears feasible and was associated with predominantly transient neurological complications. The addition of RMP-7 to carboplatin therapy appears to be at least as safe as the administration of carboplatin alone and requires further investigation as a means of chemotherapeutic dose intensification. PMID- 11386413 TI - Neurophysiological evaluation of late effects of adjuvant high-dose chemotherapy on cognitive function. AB - OBJECTIVES: To evaluate late neurotoxicity of adjuvant high-dose (HD) chemotherapy versus standard-dose (SD) chemotherapy by event-related potentials (ERP) and quantitative electroencephalography (qEEG). PATIENTS AND METHODS: From a randomized study in high-risk breast cancer patients on the efficacy of high dose versus standard-dose adjuvant chemotherapy, late effects on cognitive functioning were analyzed by neuropsychological tests. Cognitive impairment was found in 32% of the HD group, 17% of the SD group and in 9% of a control group of stage I breast cancer patients not treated with chemotherapy. In 17 consecutive patients in the HD group and 16 consecutive patients in the SD group neurophysiological tests were performed, consisting of P300 and qEEG. Results of patients treated with chemotherapy were compared with results of 14 control patients not treated with chemotherapy. All patients were tested two years after treatment. RESULTS: Asymmetry of the alpha rhythm of > or =0.5 Hz was found in 7 HD patients, 2 SD patients and in none of the control patients (p = 0.01). No differences were found between the groups with regard to frequency of alpha rhythm, alpha blocking and latency of P300. No correlation was found between neurophysiological parameters and neuropsychological performance, except for an overall relation between the P300 latencies and the total number of deviant test scores. CONCLUSION: Although the neurophysiological differences are subtle and the relation with the cognitive functioning in individual patients as measured by the neuropsychological examination is equivocal, the results suggest that there is neurophysiological support for cognitive dysfunction as a late complication of high-dose systemic chemotherapy in breast cancer. PMID- 11386414 TI - Cystic prostate metastases to the brain parenchyma: report of two cases and review of the literature. AB - Prostate carcinoma is among the leading causes of cancer death for American men. Intracranial metastases from prostatic adenocarcinoma are unusual, and cystic metastases are rare. Two cases of cystic intraparenchymal metastases from prostatic adenocarcinoma are reported. Our comprehensive literature review revealed that prostatic adenocarcinoma metastases are rarely diagnosed antemortem. If these lesions are detected early and treated with surgery and radiation therapy, survival time for patients with prostatic adenocarcinoma may be increased; however, given the rarity of these cases, routine CNS imaging of men with metastatic prostate cancer does not seem warranted. PMID- 11386415 TI - Differential expression of somatostatin receptors in medulloblastoma. AB - OBJECT: Somatostatin receptors have been found on a variety of tumours like neuroendocrine breast or brain tumours. Their detection opens new diagnostic and therapeutic paths. The aim of this work was to investigate their expression in medulloblastomas. METHODS: Using both techniques, reverse transcriptase polymerase chain reaction and immunohistochemistry, we analysed mRNA of different subtypes of somatostatin receptors in 15 medulloblastomas and the localisation of the subtype SSTR2 receptor at the cellular level in 13 medulloblastomas. All five subtypes mRNA were variably expressed in each medulloblastoma. The signal obtained after Southern blotting for SSTR2 receptor amplification was the highest as compared to the signal obtained for the other receptor subtypes. Immunostaining for SSTR2A receptor was present in every tumour specimen and was specifically located to the cellular membrane of neoplastic cells. No staining was identified at the level of peritumoral veins. CONCLUSION: The evidence of predominant expression of SSTR2 receptors in medulloblastomas opens interesting prospects for their diagnosis and therapy. PMID- 11386416 TI - Characterization of the Lactobacillus casei group and the Lactobacillus acidophilus group by automated ribotyping. AB - A total of 91 type and reference strains of the Lactobacillus casei group and the L acidophilus group were characterized by the automated ribotyping device Riboprinter microbial characterization system. The L. casei group was divided into five (C1-C5) genotypes by ribotyping. Among them, the strain of L. casei ATCC 334 was clustered to the same genotype group as most of L. paracasei strains and L casei JCM 1134T generated a riboprint pattern that was different from the type strain of L. zeae. These results supported the designation of L. casei ATCC 334 as the neotype strain, but were not consistent with the reclassification of L. casei JCM 1134T as L. zeae. The L. acidophilus group was also divided into 14 (A1-A11, B1-B3) genotypes by ribotyping. L. acidophilus, L. amylovorus, L. crispatus and L. gallinarum generated ribotype patterns that were distinct from the patterns produced by L. gasseri and L. johnsonii. This result confirmed previous data that the L. acidophilus group divided to two major clusters. Five strains of L. acidophilus and two strains of L. gasseri were correctly reidentified by ribotyping. Most strains belonging to the L. casei group and the L. acidophilus group were discriminated at the species level by automated ribotyping. Thus this RiboPrinter system yields rapid, accurate and reproducible genetic information for the identification of many strains. PMID- 11386417 TI - Characterization of an extended-spectrum class C beta-lactamase of Citrobacter freundii. AB - Citrobacter freundii GC3 is a clinical isolate which showed moderate resistance to oxyimino beta-lactams such as ceftazidime and aztreonam. This drug resistance was due to an extended-spectrum class C beta-lactamase encoded by chromosomal gene(s). The GC3 beta-lactamase showed high amino acid sequence homology to a known C. freundii beta-lactamase, i.e., 346 of 361 amino acids were identical with those of C. freundii GN346 beta-lactamase (Tsukamoto, K. et al, Eur. J. Biochem. 188, 15-22, 1990). Asp198 was the only dissimilar amino acid found in the omega loop region, known as the hot spot for extended-spectrum resistance in class C beta-lactamases (Haruta, S. et al, Microbiol. Immunol. 42, 165-169, 1998). However, Asp198 was eliminated as a cause of the extended-spectrum resistance by the substitution of Asn for Asp198. Subsequent investigation suggested that the moderate resistance to oxyimino beta-lactams is attributable to the replacement of amino acids on the enzyme's surface area, far from the active-site. Some or all of the replacements are assumed to delicately modify the active-site configuration. The GC3 beta-lactamase is the first example of an extended-spectrum class C beta-lactamase in which mutations are independent of the omega loop. PMID- 11386418 TI - Proteolytic cleavage of staphylococcal exoproteins analyzed by two-dimensional gel electrophoresis. AB - Extracellular proteases of Staphylococcus aureus are emerging as potential virulence factors that are relevant to the pathogenicity of staphylococcal infections. These proteases may also be involved in the proteolytic cleavage of other exoproteins released from this organism. To define the target exoproteins and their sites of cleavage by proteases, high-resolution two-dimensional polyacrylamide gel electrophoresis followed by N-terminal amino acid sequencing of exoprotein spots was performed. Two to three hundred exoprotein spots were detected at the early-stationary phase of cultures of S. aureus NCTC8325, and then at the late-stationary stage most of these high molecular protein spots became invisible due to further proteolytic degradation. As the result of N terminal analysis, lipase, triacylglycerol lipase, orf619 protein and orf388 protein were detected as multiple spots at the early-stationary phase. We found that these exoproteins were cleaved at 3, 7, 4 and 4 different sites, respectively, by proteases. According to the M.W. and pI of each peptide spot obtained from the gel and their matches with calculated values in addition to their N-terminal sequences, we showed that the positions of putative peptides resulted from proteolytic cleavage of these proteins. PMID- 11386420 TI - Structural characterization of the O-antigenic polysaccharide chain of Porphyromonas circumdentaria NCTC 12469. AB - Structural studies were carried out on an O-antigenic polysaccharide moiety derived from Porphyromonas circumdentaria NCTC 12469, a reference strain of Porphyromonas species. The polysaccharide chain was composed of D-glucose, D galactose, N-acetyl-D-glucosamine, and N-acetyl-D-galactosamine in a molar ratio of 1:2:1:1. On the basis of results from 1H- and 13C-NMR spectroscopic analyses including COSY, TOCSY, and HMQC experiments together with results of Smith degradation, methylation analysis, and partial acid hydrolysis, it is concluded that the polysaccharide chain has a pentasaccharide repeating unit of -->6)-beta D-Glcp-(1-->6)-beta-D-Galp-(1-->3)-beta-D-GlcpNAc-(1-->3)-beta-D-GalpNAc-(1-->. The immunoreaction between P. circumdentaria LPS and the corresponding antiserum was strongly inhibited by the pentasaccharide fragment (Glc-Gal-Gal-GlcNAc GalNAc) isolated from partial acid hydrolysis of the above polysaccharide, suggestive of O-antigen specific antibodies in the used antiserum. PMID- 11386419 TI - Intraspecies diversity of Cryptococcus albidus isolated from humans as revealed by sequences of the internal transcribed spacer regions. AB - The basidiomycetous yeast, Cryptococcus albidus, shows intraspecies diversity, but it is rarely isolated from immunocompromised patients. Nineteen strains of C. albidus, including nine clinical isolates, were re-classified by sequences of their rRNA internal transcribed spacer (ITS) regions. The nine clinical isolates were genetically diverse and included both C. albidus and C. diffluens. One clinical isolate, recovered from the blood of an AIDS patient, represented a new species. Only small differences were found in the biochemical and serological characteristics of C. albidus and C. diffluens. All isolates were sensitive to amphotericin B, but several isolates were resistant to fluconazole and itraconazole. C. albidus heterogeneity should be taken into consideration when identifying clinical isolates. PMID- 11386421 TI - Burkholderia pseudomallei interferes with inducible nitric oxide synthase (iNOS) production: a possible mechanism of evading macrophage killing. AB - Burkholderia pseudomallei is a causative agent of melioidosis, a life threatening disease which affects humans and animals in tropical and subtropical areas. This bacterium is known to survive and multiply inside cells such as macrophages. The mechanism of host defense against this bacterium is still unknown. In this study, we demonstrated that B. pseudomallei exhibited unique macrophage activation activity compared with Escherichia coli and Salmonella typhi. The mouse macrophage cell line (RAW 264.7) infected with B. pseudomallei at MOI of 0.1:1, 1:1 and 10:1 did not express a detectable level of inducible nitric oxide synthase (iNOS). Moreover, the B. pseudomallei infected cells released TNF-alpha only when they were infected with high MOI (10:1). Unlike the cells infected with B. pseudomallei, the cells infected with E. coli, and S. typhi expressed iNOS even at MOI of 0.1:1. These infected cells also released a significantly higher level of TNF-alpha at the low MOI ratio. The cells that were preactivated with IFN-gamma prior to being infected with B. pseudomallei exhibited an enhanced production of iNOS and TNF-alpha release. The increased macrophage activation activity in the presence of IFN-gamma also correlated with the restriction of the intracellular bacteria survival. Moreover, IFN-gamma also prevented cell fusion and multinucleated cell formation induced by B. pseudomallei, a phenomenon recently described by our group. Altogether, these results indicate that internalization of B. pseudomallei failed to trigger substantial macrophage activation, a phenomenon which could prolong their survival inside the phagocytic cells and facilitate a direct cell to cell spreading of B. pseudomallei to neighboring cells. PMID- 11386422 TI - Rapid detection of human fecal Eubacterium species and related genera by nested PCR method. AB - PCR procedures based on 16S rDNA gene sequence specific for seven Eubacterium spp. and Eggerthella lenta that predominate in the human intestinal tract were developed, and used for direct detection of these species in seven human feces samples. Three species of Eggerthella lenta, Eubacterium rectale, and Eubacterium eligens were detected from seven fecal samples. Eubacterium biforme was detected from six samples. It was reported that E. rectale, E. eligens, and E. biforme were difficult to detect by traditional culture method, but the nested PCR method is available for the detection of these species. This result shows that the nested PCR method utilizing a universal primer pair, followed by amplification with species-specific primers, would allow rapid detection of Eubacterium species in human feces. PMID- 11386423 TI - Identification of shiga toxin-producing Escherichia coli possessing insertionally inactivated Shiga toxin gene. AB - We have investigated the Shiga toxin genes of Shiga toxin-producing Escherichia coli (STEC) strains, using polymerase chain reaction (PCR) amplifying the full lengths of these genes. As a result, we found the Shiga toxin 2 gene which was insertionally inactivated by an insertion sequence (IS). This IS element was identical to IS1203v which has been also found in inactivated Shiga toxin 2 genes, and was inserted at the same site as in the previous paper. On the other hand, both Shiga toxin 2 genes were different (98.3% identity). These suggested that IS1203v independently inserted into each Shiga toxin 2 genes, and STEC strains possessing the insertionally inactivated Shiga toxin genes are most likely to have a wide distribution. Amplification of the full length of the Shiga toxin gene is one of the effective methods to detect the gene no matter where the IS element is included, i.e., the insertion can be reflected in the size of amplicon. PMID- 11386424 TI - Much higher risk of premalignant and malignant cervical diseases in younger women positive for HPV16 than in older women positive for HPV16. AB - Human papillomavirus (HPV), particularly HPV16, is strongly associated with premalignant lesions of the uterine cervix and cervical cancer. However, HPV infection is a common sexually transmitted disease and only a few women develop cervical cancer. Although the presence of HPV and abnormal cytology are independent risk factors for cervical diseases, implementing both tests on every woman is argued not to be recommended mainly in terms of cost-effectiveness. During a 20-month period between October, 1994, and May, 1996, cervical swabs from 207 women who were referred for colposcopy because of cervical dyskaryosis (Papanicolaou class IIIa or higher) were examined by PCR for the presence of HPV16. When these women were divided into two groups; i.e., group A consisting of women who were 44 years old or younger (n = 111), and group B consisting of women who were 45 years old or older (n = 96), the risk of having premalignant and malignant cervical diseases upon infection with HPV16 was approximately 8 times higher in group A than in group B. Thus, we conclude that HPV-testing should be implemented on every young woman with an abnormal Papanicolaou smear test. PMID- 11386426 TI - Presurgical nasoalveolar orthopedic molding in primary correction of the nose, lip, and alveolus of infants born with unilateral and bilateral clefts. AB - This addendum to the "State of the Art Dental Treatment of Predental and Infant Patients With Clefts and Craniofacial Anomalies," by Prahl-Andersen (Cleft Palate Craniofac J. 2000;37:528532), offers an extended perspective on this controversial subject. This article reviews the role of combined nasal and alveolar (nasoalveolar) molding in the primary correction of the nose, lip, and alveolus of infants born with unilateral and bilateral clefts. The background of presurgical nasoalveolar orthopedic molding, the technique, and the literature are presented. The proposed benefits of treatment from the traditional techniques of presurgical orthopedics have been shown to be unsubstantiated (Kuijpers Jagtman and Prahl, 1996). A close comparison of the proposed benefits of earlier forms of presurgical orthopedics, along with those of the current technique of nasoalveolar molding, is presented. PMID- 11386425 TI - Comparative sequences of two type 1 dengue virus strains possessing different growth characteristics in vitro. AB - The complete genome sequences of two dengue-1 virus strains having different growth characteristics (Mochizuki and A88) were compared with other published strains. The sequence analysis indicated several unique amino acid changes throughout the coding region of Mochizuki strain, mostly in envelope (E) protein. A unique amino acid, Ile-69 for Mochizuki strain at E protein resulted in the loss of an Asn-67-linked glycosylation site. A Thr substitution for Ala-114 at C protein and amino acid changes found in E, non-structural NS3, NS4a, and NS5 proteins were unique for A88 strain. These substitutions might be correlated to their different growth characteristics in vitro. PMID- 11386427 TI - Changes in speech following maxillary distraction osteogenesis. AB - OBJECTIVE: The purpose of this study was to describe changes in articulation and velopharyngeal function following maxillary distraction osteogenesis. DESIGN: This is a descriptive, post hoc clinical report comparing the performance of patients before and after maxillary distraction. The independent variable was maxillary distraction while the dependent variables were resonance, articulation errors, and velopharyngeal function. SETTING: The data were collected at a tertiary health care center in Chicago. PATIENTS: The data from pre- and postoperative evaluations of 18 maxillary distraction patients were used. OUTCOME MEASURES: The outcome measures were severity of hypernasality and hyponasality, velopharyngeal orifice size as estimated using the pressure-flow technique, and number and type of articulation errors. RESULTS: At the long-term follow-up, 16.7% exhibited a significant increase in hypernasality. Seventy-five percent of patients with preoperative hyponasality experienced improved nasal resonance. Articulation improved in 67% of patients by the 1-year follow-up. CONCLUSIONS: In a predominately cleft palate population, the risk for velopharyngeal insufficiency following maxillary distraction is similar to the risk observed in Le Fort I maxillary advancement. Patients being considered for maxillary distraction surgery should receive pre- and postoperative speech evaluations and be counseled about risks for changes in their speech. PMID- 11386429 TI - "Only skin deep": health, resilience, and craniofacial care. AB - The literature on psychological and social responses to cleft palate and craniofacial conditions focuses on the challenges and limitations posed by biological factors, cognitive status, and social stigma. In spite of the various challenges experienced by persons with these conditions, many individuals have rich and satisfying lives. This paper develops a theoretical perspective to elucidate factors that account for health, life success, and resiliency in persons with craniofacial conditions. Three steps are proposed to change how craniofacial conditions are envisioned by clinicians and researchers: (1) creating optimism and positivity around the time of birth and diagnosis, (2) asking children and parents new and different questions, and (3) launching a new craniofacial social science model for research built around understanding resilience and health. PMID- 11386428 TI - Correction of coronal suture synostosis using suture and dura mater allografts in rabbits with familial craniosynostosis. AB - OBJECTIVE: Resynostosis following surgical correction of craniosynostosis is a common clinical correlate. Recent studies suggest that the dura mater is necessary to maintain suture patency. It has also been hypothesized that dura mater from synostotic individuals may provide aberrant biochemical signals to the osteogenic fronts of the calvaria, which result in premature suture fusion and subsequent resynostosis following surgery. This study was designed to test this hypothesis by surgically manipulating the coronal suture and dura mater in rabbits with familial craniosynostosis to prevent postsurgical resynostosis. DESIGN: Craniofacial growth and histomorphometric data were collected from 129 rabbits: 72 normal controls and 57 rabbits with bilateral coronal suture synostosis (15 unoperated on controls; 13 surgical controls; 9 dura mater transplant only; 10 suture transplant only; and 10 suture and dura mater transplant). At 10 days of age, all rabbits had radiopaque amalgam markers placed on either side of the coronal, frontonasal, and anterior lambdoidal sutures. At 25 days of age, 42 synostosed rabbits had a 3 to 5-mm wide coronal suturectomy. Coronal sutures and/or underlying dura mater allografts were harvested from same aged, wild-type, isohistogenic control rabbits and transplanted onto the dura mater of synostosed host rabbits. Serial radiographs were taken at 10, 25, 42, and 84 days of age, and the suturectomy sites were harvested at 84 days of age in 44 rabbits and serially sectioned for histomorphometric examination. RESULTS: Results revealed that cranial vault growth was significantly (p < .05) improved following surgical release of the fused coronal suture compared with synostosed rabbits who were not operated on but was still significantly different (p < .05) from that of normal control rabbits. By 84 days of age, significant (p < .05) differences were noted in calvarial suture marker separation, cranial vault shape indices, and cranial base angles between rabbits with and without dura mater allografts, probably as a result of resynostosis of the suturectomy site or suture-only allografts. Qualitative histological examination revealed that at 84 days of age rabbits with suture and dura allografts had patent coronal sutures, suture-only allografts had fused coronal sutures with extensive endosteal hyperostosis, dura mater-only allografts had some new bone in the suturectomy site that resembled rudimentary osteogenic fronts, and suturectomy controls had extensive endosteal bone formation and resynostosis of the suturectomy site. Significantly (p < .05) more bone was found in the suturectomy sites of rabbits without dura mater allografts compared with rabbits with dura mater allografts. CONCLUSIONS: Results support the initial hypothesis that normal dura mater allografts will maintain suture or suturectomy site patency and allow unrestricted craniofacial growth. However, it is still unclear whether the dura mater from normal rabbits was providing biochemical signals to the transplanted sutures or suturectomy sites or simply acting as a barrier to prevent abnormal biochemical signals from the dura mater of synostosed rabbits from reaching the calvaria. The clinical and therapeutic implications of these procedures are discussed. PMID- 11386430 TI - Resiliency and success in adults with Moebius syndrome. AB - OBJECTIVE: This study asked selected mature individuals with Moebius syndrome to discuss the sources of strength and resiliency that allowed them to achieve professional and personal success. DESIGN: Adults with Moebius syndrome were selected in a nonrandom manner based primarily on their affiliation with the Moebius Syndrome Foundation. Following a letter from the author describing the objective, the subjects were interviewed by telephone and were encouraged to respond at length. PARTICIPANTS: Eighteen adults (aged 29 to 70 years) responded. All were or had been gainfully employed in a variety of professional or vocational positions. RESULTS: The respondents reported the following as major sources of resiliency and success: family support, faith, humor, sense of self, special skills, determination, and networking. CONCLUSIONS: The recognition and reinforcement of strengths and resiliences in younger patients may help maximize their professional and personal success as adults. PMID- 11386431 TI - Unique outcomes and positive contributions associated with facial difference: expanding research and practice. AB - Current literature on the psychosocial outcomes of living with a craniofacial difference (CFD) focuses primarily on deficits. In this paper, an alternative perspective is explored in a pilot study of 11 parents of children with CFDs and 11 affected adults reflecting on their experiences. Mixing qualitative and quantitative methodologies, this pilot study begins to demonstrate the value of exploring contributional outcomes related to CFDs. The findings in this study include positive outcomes in terms of communication abilities, service to others, observational skills, inner strength, abilities to question society, a valued social circle, and perceptions of being normal because of the difference. The findings also reveal that approximately half of the parents and affected adults would not choose to remove the experience of facial difference from their lives if they have such an option. Implications for future research, practice, and ethical and policy considerations are discussed. PMID- 11386432 TI - Facing up to stigma: workplace and personal strategies. AB - This paper examines some of the influences that contribute to health, life success, and resiliency in persons with craniofacial conditions. Those influences are examined from the perspective of the author, a 50-year-old man with cleft palate, right microtia, and hemifacial microsomia. A biosketch describes the author as one example of someone with craniofacial conditions who has also achieved significant personal and professional success. Central to that success were the influences of health care providers and parents as well as other people with craniofacial conditions. Specific attention is given to the benefits of a team approach to the care of patients with craniofacial conditions; the role of health care professionals in fostering the optimism that can mobilize the family for action; the importance of acknowledging the intransigence of stigma in our culture; the value of practical advice that affected individuals can offer one another. PMID- 11386433 TI - Using psychological assessment and therapeutic strategies to enhance well-being. AB - OBJECTIVE: To provide an overview of the psychological assessment; results from studies examining psychological issues among individuals with craniofacial anomalies (CFA) and other chronic conditions; resilience; and therapeutic strategies to enhance psychological well-being. CONCLUSIONS: The literature on chronic conditions and findings from studies with people having CFA and their families demonstrate a range of effective adaptation patterns and strategies to enhance issues having an impact on quality of life. PMID- 11386434 TI - Deficits versus strengths: ethics and implications for clinical practice and research. AB - A shift in emphasis from deficits to strengths to promote health and well-being in patients with congenital and acquired craniofacial conditions (CFC) is appropriate given the chronic, "incurable" nature of CFC. Personal narratives are a valuable starting point for discovering sources of resilience. This paper explores such a shift by considering two areas of ethical concern suggested by patient narratives: informed consent and the physician-patient relationship. Both areas contain pitfalls and opportunities. The powerful emotions these patients bring with them argue for caution in medical decision making. Attention to the individual's psychosocial adjustment should always supplement surgical evaluations. Because of the inequality of the physician-patient relationship, care should be taken to use this power in positive ways. The relationship between patient and surgeon is particularly charged and may be an important source of support, information, hope, and advice for patients. The changing health care system threatens the physician-patient relationship, but the rise of alternative medicine suggests patients continue to value relationships. Relationships are critical for individuals with CFC, who experience social rejection because of the fundamental importance of face in human interactions. Future research directions should include long-term outcome studies on patients receiving modern craniofacial team care, qualitative research on resilience in patients with positive life adjustment, and development of a conceptual framework and research methodology for understanding quality of life of individuals with CFC. An emphasis upon strengths rather than defects will have implications for the structure of craniofacial teams, the care that is provided, and allocation of resources. PMID- 11386435 TI - Normative values of craniofacial measurements in idiopathic benign macrocephalic children. AB - OBJECTIVE: To determine normal standard values at each age for head circumference; inner canthal distance; outer canthal distance; near interpupillary distance; distant [far] interpupillary distance; canthal index; and circumference-interorbital index specific for the age, sex, and race in 7- to 16-year-old idiopathic benign macrocephalic male and female children. METHODS: The measurements of head circumference were obtained in 8736 children (4591 boys and 4145 girls) in the city center of Malatya. From these, macrocephalic children (boys = 103, girls = 93) were chosen and invited to our clinic for detailed examination. Mean values for each parameter were obtained at each age from 7 to 16 years. RESULTS: The mean age for male and female children were 11.87 +/- 2.87 and 11.72 +/- 2.75 years, respectively. The overall mean values for head circumference, inner canthal distance, outer canthal distance, near interpupillary distance, distant interpupillary distance, canthal index and circumference-interorbital index in boys were found to be 57.43 +/- 1.46 cm, 31.90 +/- 2.41 mm, 89.29 +/- 4.34 mm, 58.79 +/- 2.97 mm, 62.10 +/- 3.23 mm, 35.73 +/- 2.24, and 5.56 +/- 0.44, respectively. In girls, these values were 56.57 +/- 1.24 cm, 31.45 +/- 2.65 mm, 87.71 +/- 4.11 mm, 58.16 +/- 3.46 mm, 61.26 +/- 3.66 mm, 35.84 +/- 2.31, and 5.56 +/- 0.43, respectively. CONCLUSION: Normal values of craniofacial measurements for idiopathic benign macrocephalic healthy children are useful in early identification of some craniofacial syndromes, congenital or posttraumatic telecanthus, epicanthus, and hypo-hypertelorism and of planning surgical intervention. We suggest that the comparison of craniofacial dimensions of macrocephalic healthy children must be performed with normal standards specific for age as well as sex and race. PMID- 11386436 TI - Three-dimensional Z-plasty in the correction of the unilateral cleft lip nasal deformity. AB - OBJECTIVE: Numerous methods have been introduced for correction of the cleft lip nasal deformities, but no single procedure has given sufficiently satisfactory results to provide a surgical standard. Much effort has been put on restoring cartilaginous structures using alar cartilage modification and suspension. But even after the cartilage framework is repositioned well, redundant alar webbing is still unsightly and frequently conspicuous. This paper presents a procedure combining the usual open rhinoplasty technique and three-dimensional Z-plasty in an external approach to remove the alar web and to lengthen the columella at the same time. METHODS: Open rhinoplasty was performed in 26 consecutive patients with unilateral cleft lip nasal deformity from 1991 to 1996. We used an open rhinoplasty approach, which is a combination of the usual infracartilaginous incision on the noncleft side and a small triangular flap on the cleft side. RESULTS AND CONCLUSIONS: This external rhinoplasty incision can provide a wide surgical field for the handling of the whole cartilage framework. The three dimensional Z-plasty utilizing redundant alar skin not only removes the alar web but also helps lengthen the columella and provides soft tissue to the nasal vestibule. This technique is easy, and the postoperative result is very consistent. Our open rhinoplasty approach has several advantages and can be a useful approach in the correction of the unilateral cleft lip nasal deformity. PMID- 11386437 TI - La maladie de Grisel: spontaneous atlantoaxial subluxation. AB - OBJECTIVE: "La maladie de Grisel" (Grisel's syndrome) is a spontaneously occurring atlantoaxial subluxation with torticollis. We present a case of atlantoaxial subluxation occurring in a 20-year period of pharyngoplasty surgery. The occurrence of a "spontaneous" atlantoaxial subluxation after oral cavity or pharynx operations is rare. Because some neck pain and stiffness are commonly seen after these kinds of operations, we would like to draw attention to this unusual complication. Symptoms associated with a torticollis after an operation in the oral cavity or pharynx requires additional investigation to exclude this rare complication. A review of the available literature concerning etiology and treatment of la maladie de Grisel is presented. PMID- 11386438 TI - Airway obstruction due to a rudimentary premaxilla in holoprosencephaly. AB - OBJECTIVE: We present two infants with holoprosencephaly and clefts of the primary palate, who developed nasal airway obstruction due to a previously undocumented cause: a ball-valve-like action of a rudimentary premaxilla, which was extremely mobile on a soft tissue stalk. In such patients, the importance of intranasal examination to identify a rudimentary premaxilla, in order to preserve it, and if necessary, surgically stabilize it, is illustrated. PMID- 11386439 TI - Higher incidence of twins in infants with Pierre Robin sequence. PMID- 11386440 TI - 'An unpredictable and rampant virus'. PMID- 11386441 TI - Suspension ordered following farriery charge. PMID- 11386442 TI - Reprimand after breach of greyhound racing rules. PMID- 11386443 TI - Spread of foot-and-mouth disease from the burning of animal carcases on open pyres. AB - Preliminary results indicate that no evidence has been found to support the spread of FMD virus from the burning of animal carcases on open pyres. This finding is subject to a number of assumptions, and is based on a limited number of case studies. PMID- 11386444 TI - Morbillivirus in common seals stranded on the coasts of Belgium and northern France during summer 1998. AB - Sixteen common seals (Phoca vitulina) were stranded on the Belgian and northern French coasts during the summer of 1998. Eleven (10 pups and one adult) were sampled for histopathological, immunohistochemical, serological, bacteriological, parasitological and virological investigations. The main gross findings were severe emaciation, acute haemorrhagic enteritis, acute pneumonia, interstitial pulmonary emphysema and oedema, and chronic ulcerative stomatitis. Microscopical lung findings were acute to subacute pneumonia with interstitial oedema and emphysema. Severe lymphocytic depletion was observed in lymph nodes. Severe acute to subacute meningoencephalitis was observed in one animal. Specific staining with two monoclonal antibodies directed against canine distemper virus (CDV) and phocine distemper virus was observed in a few lymphocytes in the spleen and lymph nodes of three seals. Anti-CDV neutralising antibodies were detected in sera from six animals. Seven of the seals were positive by reverse transcriptase-PCR for the morbillivirus phosphoprotein gene. The lesions observed were consistent with those in animals infected by a morbillivirus, and demonstrated that distemper has recently recurred in North Sea seals. PMID- 11386445 TI - Study of the behaviour, digestive efficiency and gut transit times of crib-biting horses. AB - The spontaneous behaviour and the apparent digestibility of dry matter and fibre and transit times of digesta were compared in four normal horses and four crib biters. A technique was developed for measuring total gut transit times (TGTT) by using single-stool analysis of the passage of radio-opaque polyethylene markers. Longer TGTT were recorded in the crib-biters than in the normal horses but the orocaecal transit times did not differ. The crib-biters rested less than the normal horses. PMID- 11386446 TI - Evaluation of the dosage of ivermectin in falcons. AB - Twelve groups of falcons, each containing three female gyrfalcon-peregrine falcon hybrids (Falco rusticolus x Falco peregrinus) were injected intramuscularly with a single dose of ivermectin ranging from 0.2 mg/kg to 11 mg/kg bodyweight, and a control group was injected with water. Doses of ivermectin between 0.2 and 5 mg/kg failed to produce clinical signs of illness in the birds. Four birds which received either 6, 7 or 8 mg/kg showed slight clinical signs, and all the birds receiving 9 to 11 mg/kg showed more or less severe clinical signs of anorexia, apathy and sedation. Slight changes in the mean plasma activities of aspartate aminotransferase, alanine aminotransferase and alkaline phosphatase (AP) were detected in the group dosed with 5 mg/kg, and higher dosages caused marked changes in these enzymes as well as in the mean plasma activity of lactate dehydrogenase. The mean activity of AP decreased, and the activities of the other enzymes increased. A dosage of 2 to 3 mg/kg ivermectin is recommended as a safe and effective antiparasitic drug for falcons and it has been used successfully to treat infestations of Serratospiculum species. PMID- 11386447 TI - Relative resistance of pigs to infection by natural aerosols of FMD virus. PMID- 11386448 TI - Relative risks of the uncontrollable (airborne) spread of FMD by different species. PMID- 11386449 TI - European policies for the control of exotic diseases. PMID- 11386450 TI - FMD control strategies. PMID- 11386451 TI - FMD control strategies. PMID- 11386452 TI - FMD control strategies. PMID- 11386453 TI - Burring cheek teeth in rabbits. PMID- 11386454 TI - Privilege to dispense. PMID- 11386455 TI - Equine grass sickness website. PMID- 11386456 TI - COPD study. PMID- 11386457 TI - Structure of radiotherapy in Japan. PMID- 11386458 TI - The future demand for and structural problems of Japanese radiotherapy. AB - BACKGROUND: Recently, as the number of elderly people in Japan is growing, so is the number of new cancer cases. The number of patients treated with radiotherapy is therefore also on the increase, so that it is important to estimate the future demand for radiotherapy and to make preparations for it. METHODS: All the surveys were conducted for 106 facilities selected randomly out of 556 radiotherapy facilities in Japan. To obtain trends in the number of new cancer patients treated with radiotherapy in Japan, we conducted a survey with a self administered mail questionnaire designed to obtain the number of new patients treated with radiotherapy for each year of the past decade (1990-99). The future number of new patients treated with radiotherapy was estimated from the data thus obtained. To investigate structural problems of Japanese radiotherapy, surveys about the number of treatment machines and full-time equivalent (FTE) radiation oncologists were conducted according to data from the Japanese Society for Therapeutic Radiology and Oncology (JASTRO) structure survey and the Patterns of Care Study (PCS). We also compared the structure of Japanese radiotherapy with that in the USA. RESULTS: The number of patients treated with radiotherapy has increased for every institutional stratum, with an overall increase of 1.4-fold over the past 10 years in Japan. It is estimated that the number of cancer patients treated with radiotherapy will reach 190 000 in 2015. In Japanese non academic institutions, less than one FTE radiation oncologist has been managing many of these patients. In both equipment and manpower, academic institutions exceed nonacademic institutions. CONCLUSION: The future demand for Japanese radiotherapy will grow substantially, so that it is of vital importance to prepare for it. Specifically, the number of FTE radiation oncologists must be increased. PMID- 11386459 TI - Influence of thoracic radiotherapy on exhaled nitric oxide levels in patients with lung cancer. AB - BACKGROUND: To determine the physiological role of exhaled nitric oxide (NO) in patients with lung cancer. METHODS: We investigated changes in exhaled NO levels in 29 patients undergoing thoracic radiation therapy with or without chemotherapy. The exhaled NO level was assessed using a chemiluminescence analyzer. RESULTS: The level of exhaled NO was higher in patients with lung cancer before treatment than in controls. With radiotherapy, the exhaled NO level decreased for patients undergoing 40 Gy irradiation and post-radiotherapy. However, five patients showed elevated levels of exhaled NO three times or more than that before radiotherapy. Three of these patients showed signs of radiation pneumonitis. However, none of the other patients showed signs of radiation pneumonitis (p = 0.002). CONCLUSION: Radiation therapy can lower exhaled levels of NO and the levels of exhaled NO may be a useful index for the early prediction of radiation pneumonitis. PMID- 11386460 TI - Pilot study of local radiotherapy for portal vein tumor thrombus in patients with unresectable hepatocellular carcinoma. AB - BACKGROUND: Patients suffering from hepatocellular carcinoma (HCC) with portal vein tumor thrombus (PVTT) generally have a poor prognosis. We therefore conducted a prospective pilot trial of combined transcatheter arterial chemoembolization (TACE) and local radiotherapy (RT) for PVTT in unresectable HCC. The aim of the study was to investigate the efficacy and toxicity of this preliminary trial regime and to explore RT guidelines for cirrhosis. METHODS: Eight patients with unresectable HCC accompanied by first branch PVTT were entered into the study from February 1998 to December 1999. TACE was performed using Lipiodol, epirubicin hydrochloride and mytomycin followed by gelatin sponge cubes. RT was started 10-14 days following TACE. A total delivered dose of 60 Gy was given as daily 2 Gy fractions, with the clinical target volume defined as PVTT only. We observed a relationship between deterioration of liver function and the percent volume of the total liver receiving a dose exceeding 30 Gy (V30). RESULTS: An objective response was observed in three of the eight patients. However, on follow-up angiograms the protrusion of PVTT into the main portal trunk was decreased in all cases. Deterioration of liver function was observed in all patients with V30 >40%. CONCLUSION: It is possible that this combined therapy prevents PVTT from spreading to the main trunk and that indicates a further benefit of TACE. Our results indicate that V30 constitutes a predictive test for the development of liver failure. More detailed evaluations of liver function and determination of the safe irradiation volume are necessary. PMID- 11386461 TI - Node-positive mucosal gastric cancer: a follow-up study. AB - BACKGROUND: Lymph node metastasis from mucosal gastric carcinoma is rare and the prognosis of the patients has seldom been reported. METHODS: Forty-five patients with node-positive mucosal gastric cancer were studied. They accounted for 2.5% of 1770 patients with mucosal gastric cancer who underwent gastrectomy with lymphadenectomy at the National Cancer Center Hospital, Tokyo. The clinicopathological features were studied and the current clinical status was sought. RESULTS: The majority of patients (87%) were treated with D2 lymphadenectomy. The metastasis was confined to the perigastric nodes (pN1 by Japanese classification) in 30 patients (67%). The number of positive nodes was less than seven (pN1 by TNM) in 42 patients (93%). Two patients had para-aortic nodal metastasis. The median follow-up period was 11 years. Four patients died of definite or possible recurrent disease and the disease-specific 5- and 10-year survival rates were 95 and 89%, respectively. CONCLUSIONS: Although nodal metastasis is an important prognostic factor for gastric cancer, the prognosis was excellent as long as the primary tumor was confined to the mucosa and was treated with gastrectomy and lymphadenectomy. PMID- 11386462 TI - A family of multiple endocrine neoplasia type 2A with the RET proto-oncogene mutation in codon 618 (Cys-->Arg). AB - Multiple endocrine neoplasia type 2 (MEN-2) is a hereditary syndrome characterized by medullary thyroid carcinoma (MTC), pheochromocytoma and hyperplasia or adenoma of the parathyroid gland with hyperparathyroidism. Recent genetic studies have identified the presence of germline missense mutations in the RET proto-oncogene in almost 100% of MEN-2 patients. We report here three generations of one MEN-2 family with rare missense mutation at codon 618 (Cys- >Arg) of the RET proto-oncogene. The first patient was surgically treated at the age of 63 years but died of bone metastasis. His two children (29-year-old daughter and 25-year-old son) were treated surgically for MTC and neck lymph node metastasis. Germline mutations of the RET proto-oncogene of these three MTC patients and two children of the 29-year-old daughter (9-year-old female and 7 year-old male) were examined. Three MTC patients and the 9-year-old female possessed the mutation. The phenotype of the family with this rare point mutation of the RET proto-oncogene is reported. PMID- 11386463 TI - Intestinal perforation due to metastasis of breast carcinoma, with special reference to chemotherapy: a case report. AB - We report a case of small-bowel perforation due to metastatic carcinoma of the breast during chemotherapy. Partial resection of the small intestine and primary anastomosis were performed. Although the patient made a good recovery from panperitonitis, she died of the disease on the 55th postoperative day. Since perforation during chemotherapy results in an extremely poor prognosis, special caution during chemotherapy is needed for patients with possible gastrointestinal involvement with tumor. PMID- 11386464 TI - Intestinal metastasis causing intussusception in a patient treated for osteosarcoma with history of multiple metastases: a case report. AB - Intestinal intussusception caused by metastatic tumors is a very rare condition. Preoperative diagnosis is not easy because of the condition's rarity and because of mild abdominal physical presentation. We report on a patient with osteosarcoma who suffered from abdominal pain and emesis during the period of autologous peripheral blood stem cell transplantation. He had undergone tumor excision and radiotherapy several times prior to autologous peripheral blood stem cell transplantation because of multiple metastases. Intestinal metastasis was suspected initially by computed tomographic scan and sonogram and was proved by surgical resection and pathological findings. Clinicians caring for pediatric patients with osteosarcoma with a history of multiple metastases should consider the possibility of intestinal metastases when equivocal abdominal symptoms develop after intensive chemotherapy. PMID- 11386465 TI - Atypical bone metastasis and radiation changes in a colon cancer: a case report and a review of the literature. AB - We report a unique case of skeletal metastasis with prominent soft tissue extension from a colon cancer in a 44-year-old male patient. Four years after the diagnosis of colon cancer, plain radiographic examinations revealed a prominent soft tissue mass associated with cortical destruction of the right femur. Palliative radiotherapy of the right femur was performed. After completing radiotherapy, massive remineralization was seen in the soft tissue component. With the assistance of pre- and post-radiotherapy computerized tomography images, obtained in the process of planning the radiotherapy, we reached the conclusion that in our case the pre-radiotherapy finding was an extreme case of eccentric expansion of the bony cortex due to the outgrowth of bony metastasis. Retrospectively, the initial prominent soft tissue mass associated with the cortical destruction seems to have been a protruding bone metastasis, extending from the medial aspect of the right femur. This peculiar feature with remineralization after radiotherapy should help explain the phenomenon of atypical bony metastasis from a radiological perspective. PMID- 11386467 TI - Colon cancer mortality rates by prefectures in Japan. PMID- 11386466 TI - Case report: hand-foot syndrome induced by the oral fluoropyrimidine S-1. AB - Hand-foot syndrome (HFS) is a relatively common side effect of fluorouracil (5 FU) chemotherapy that has also been associated with the oral fluoropyrimidine capecitabine. Interestingly, HFS is virtually unknown to result from treatment with UFT, a combination of tegafur and uracil. Tegafur is a prodrug of 5-FU and is a component of S-1, another oral fluoropyrimidine active in a variety of solid tumors. We know of only one previously described case of S-1-induced HFS and the case reported here is the first to provide full documentation of this occurrence. The pathophysiology of chemotherapy-induced HFS remains unknown and very little pathological information is available. Treatment consists of topical emollient therapy, although pyridoxine has occasionally been beneficial. The study of HFS may provide an important insight into the pharmacology of fluoropyrimidines and allow for effective preventive strategies for this side effect of chemotherapy. PMID- 11386468 TI - Swallowing and tongue function following treatment for oral and oropharyngeal cancer. AB - This study examined tongue function and its relation to swallowing in 13 subjects with oral or oropharyngeal cancer treated with primary radiotherapy +/- chemotherapy and 13 age- and sex-matched control subjects. Measures of swallowing and tongue function were obtained using videofluoroscopy, pretreatment and 2 months posttreatment. Maximum isometric strength and endurance at 50% of maximum strength were obtained with the Iowa Oral Performance Instrument (IOPI). Control subjects were tested once. All subjects with head and neck cancer were evaluated pretreatment and 2 months posttreatment. No significant differences were found for the tongue function measures pre- and 2 months posttreatment in the group with head and neck cancer. Significantly higher tongue strength was observed in the control than in the group with head and neck cancer both pre- and posttreatment. No significant differences were found for the 2 groups for tongue endurance measures. Significant correlations of tongue strength and endurance and some swallow measures were found pre- and posttreatment for the group with head and neck cancer and for the control group. These correlations included oral and pharyngeal temporal swallow measures and oropharyngeal swallow efficiency. Pretreatment differences between the 2 groups in tongue strength were likely related to tumor bulk, pain, and soreness. Two-month posttreatment differences were likely related to radiation +/- chemotherapy changes to the oral and pharyngeal mucosa. This study provides support for the hypothesis that tongue strength plays a role in oropharyngeal swallowing, particularly related to the oral phase of the swallow. PMID- 11386469 TI - Respiratory control in stuttering speakers: evidence from respiratory high frequency oscillations. AB - This study tested the hypothesis that, in stuttering speakers, relations between the neural control systems for speech and life support, or metabolic breathing, may differ from relations previously observed in normally fluent subjects. Bilaterally coherent high-frequency oscillations in inspiratory-related EMGs, measured as maximum coherence in the frequency band of 60-110 Hz (MC-HFO), were used as indicators of participation by the brainstem controller for metabolic breathing in 10 normally fluent and 10 stuttering speakers. In all controls and most stuttering subjects, MC-HFO for speech was higher than or comparable to MC HFO for deep breathing. For 4 stuttering subjects, higher MC-HFO was observed for speech than for deep breathing. Comparison of deep breathing to a speechlike breathing task yielded similar results. No relationship between MC-HFO during speech and severity of disfluency was observed. We conclude that in some stuttering speakers, the relations between respiratory controllers are atypical, but that high participation by the HFO-producing circuitry in the brainstem during speech is not sufficient to disrupt fluency. PMID- 11386470 TI - A positron emission tomography study of silent and oral single word reading in stuttering and nonstuttering adults. AB - Over the last decade positron emission tomography (PET) has been used extensively for the study of language and other cognitive and sensorimotor processes in healthy and diseased individuals. In the present study, [15O]H2O PET scanning was used to investigate the lateralization and functional distribution of cortical and subcortical activity involved in single word reading in stuttering and nonstuttering individuals. Ten right-handed male stuttering adults and matched nonstuttering individuals were instructed to read individually presented single words either silently or out loud. Subtraction of functional brain images obtained during each of the two reading tasks, and during a non-linguistic baseline task, was used to calculate within-group and between-group differences in regional cerebral blood flow by means of statistical parametric mapping. Increased activation in the left anterior cingulate cortex (ACC) was observed during silent reading in the stuttering speakers but not in the nonstuttering group. Because of the hypothesized role of the ACC in selective attention and covert articulatory practice, it is suggested that the observed increased ACC activation in the stuttering individuals reflects the presence of cognitive anticipatory reactions related to stuttering. During the oral reading task, within-group comparisons showed bilateral cortical and subcortical activation in both the stuttering and the nonstuttering speakers. Between-group comparisons showed a proportionally greater left hemisphere activation in the nonstuttering speakers, and a proportionally greater right hemisphere activation in the stuttering individuals. The results of the present study provide qualified support for the hypothesis that stuttering adults show atypical lateralization of language processes. PMID- 11386471 TI - Improving the positive predictive value of screening for developmental language disorder. AB - In a previous study, we reported the results of an early language screening program in which 306 children were screened using a parent-report questionnaire sent through the mail (Klee et al., 1998). A sample of the children screened were given clinical evaluations within a month of screening (n = 64) and again 1 year later (n = 36). Although the screening program correctly identified 91% of 2-year olds with language delay, it produced a large number of over-referrals. In the present study we examine a revised screening criterion designed to reduce the number of false positives. The revised criterion generated fewer positive screens overall than the original and resulted in improved specificity (96% vs. 87%) and positive predictive value (77% vs. 51%), while maintaining the high sensitivity (91%) and negative predictive value (98%) of the original criterion. We also propose a screening score based on the new criterion, designed to inform the process of deciding which children to bring in for further evaluation. PMID- 11386472 TI - Tense and temporality: a comparison between children learning a second language and children with SLI. AB - This study compares the morphosyntax of children with SLI to the morphosyntax of children acquiring a second language (L2) to determine whether the optional infinitive phenomenon (M. Rice, K. Wexler, & P. Cleave, 1995; K. Wexler, 1994) is evident in both learner groups and to what extent cross-learner similarities exist. We analyzed spontaneous production data from French-speaking children with SLI, English-speaking L2 learners of French, and French-speaking controls, all approximately 7 years old. We examined the children's use of tense morphology, temporal adverbials, agreement morphology, and distributional contingencies associated with finiteness. Our findings indicate that the use of morphosyntax by children with SLI and by L2 children has significant similarities, although certain specific differences exist. Both the children with SLI and the L2 children demonstrate optional infinitive effects in their language use. These results have theoretical and clinical relevance. First, they suggest that the characterization of the optional infinitive phenomenon in normal development as a consequence of very early neurological change may be too restrictive. Our data appear to indicate that the mechanism underlying the optional infinitive phenomenon extends to normal (second) language learning after the primary acquisition years. Second, they indicate that tense-marking difficulty may not be an adequate clinical marker of SLI when comparing children with impairment to both monolingual and bilingual peers. A more specific clinical marker would be more effective in diagnosing disordered populations in a multilingual context. PMID- 11386473 TI - Specific language impairment in Swedish: the status of verb morphology and word order. AB - Several competing proposals have been offered to explain the grammatical difficulties experienced by children with specific language impairment (SLI). In this study, the grammatical abilities of Swedish-speaking children with SLI were examined for the purpose of evaluating these proposals and offering new findings that might be used in the development of alternative accounts. A group of preschoolers with SLI showed lower percentages of use of present tense copula forms and regular past tense inflections than normally developing peers matched for age and younger normally developing children matched for mean length of utterance (MLU). Word order errors, too, were more frequent in the speech of the children with SLI. However, these children performed as well as MLU-matched children in the use of present tense inflections and irregular past forms. In addition, the majority of their sentences containing word order errors showed appropriate use of verb morphology. None of the competing accounts of SLI could accommodate all of the findings. In particular, these accounts--or new alternatives-must develop provisions to explain both the earlier acquisition of present tense inflections than past tense inflections and word order errors that seem unrelated to verb morphology. PMID- 11386474 TI - Nonword repetition performance in school-age children with and without language impairment. AB - This study examined nonword repetition performance in a population-based sample of school-age children. A total of 581 second graders who were participating in a longitudinal, epidemiologic investigation of specific language impairment (SLI) were administered the Nonword Repetition Task (NRT) developed by Dollaghan & Campbell (1998). Performance was examined according to second-grade diagnostic category, presence/absence of language impairment, and treatment status. Results indicated that children with language impairment, as well as those in intervention, exhibited deficient nonword repetition skills compared to normal language controls. Findings also confirmed that the NRT is a culturally nonbiased measure of language processing. Results from likelihood ratio analyses indicated that NRT performance, though not sufficient on its own, may provide a useful index to assist in ruling in or ruling out language disorder. PMID- 11386475 TI - Prescribed clinician-fit versus as-worn coupler gain in a group of elderly hearing-aid wearers. AB - This study reports the prescribed, clinician-fit, coupler gain and the user adjusted, as-worn coupler gain measured in 55 adults ranging in age from 60 to 83 years (M = 72.2 years). All participants were fit with linear, output-limiting compression, Class D circuits in full-concha, in-the-ear (ITE) shells. The NAL-R prescription rationale was used to generate target real-ear insertion gain (REIG) and coupler gain values. The clinician-fit gain was measured when the hearing aid was dispensed initially and was found to be a close match to the prescribed coupler gain. Both clinician-fit and as-worn gain were measured subsequently at approximately 2 weeks, 1 month, 6 months, and 1 year after the initial fitting. As-worn gain was measured as soon as the participant returned to the clinic for one of the follow-up visits by simply removing the hearing aids and placing them in the test chamber without any adjustments in volume control. At each follow-up session, the clinician then inspected the hearing aids, evaluated the instruments electroacoustically, readjusted the volume control to the setting used to match the prescribed gain in the initial fit, and measured the clinician-fit coupler gain once again. Results revealed that, despite the capability of the hearing aid to achieve coupler gain that is a close match to the prescribed gain, these users consistently selected as-worn gain that was generally 6-9 dB below that prescribed by the NAL-R formula. Of this 6-9 dB disparity, however, as much as 3 6 dB could be due to binaural summation effects not taken into consideration in the NAL-R prescriptive formula. In addition, 5.4% of the time, the hearing aids were found to be in less than ideal operating condition when removed for the as worn gain measurements (e.g., weak or dead battery, cerumen occluding the sound bore, telecoil switch in the incorrect position). PMID- 11386476 TI - Effects of methylphenidate (Ritalin) on auditory performance in children with attention and auditory processing disorders. AB - A double-blind, placebo-controlled study was used to investigate the effects of methylphenidate (Ritalin) on tests of auditory processing in children diagnosed with both Attention Deficit Hyperactivity Disorder (ADHD) and Central Auditory Processing Disorder (CAPD). Thirty-two subjects received three Central Auditory Processing (CAP) tests and the Auditory Continuous Performance Test (ACPT), a measure of attention/impulsivity, at two separate test sessions: once when medicated with Ritalin and once when nonmedicated (placebo). Sixteen subjects were assigned randomly to receive their medication first and 16 to receive the placebo first. A counterbalanced 2 x 2 mixed factorial analysis of variance was conducted for each of the four dependent variables: Staggered Spondaic Word (SSW), Phonemic Synthesis (PS), Speech-in-Noise (SN), and ACPT measures. Analyses revealed that Ritalin did not have a significant effect on any of the three CAP measures. However, ACPT performance was significantly better (p < .000) for the Ritalin versus placebo condition. PMID- 11386477 TI - The relation between stimulus context, speech audibility, and perception for normal-hearing and hearing-impaired children. AB - In this study, the influence of stimulus context and audibility on sentence recognition was assessed in 60 normal-hearing children, 23 hearing-impaired children, and 20 normal-hearing adults. Performance-intensity (PI) functions were obtained for 60 semantically correct and 60 semantically anomalous sentences. For each participant, an audibility index (AI) was calculated at each presentation level, and a logistic function was fitted to rau-transformed percent-correct values to estimate the SPL and AI required to achieve 70% performance. For both types of sentences, there was a systematic age-related shift in the PI functions, suggesting that young children require a higher AI to achieve performance equivalent to that of adults. Improvement in performance with the addition of semantic context was statistically significant only for the normal-hearing 5-year olds and adults. Data from the hearing-impaired children showed age-related trends that were similar to those of the normal-hearing children, with the majority of individual data falling within the 5th and 95th percentile of normal. The implications of these findings in terms of hearing-aid fitting strategies for young children are discussed. PMID- 11386478 TI - Effects of age and hearing sensitivity on the use of prosodic information in spoken word recognition. AB - It is well known that spoken words can often be recognized from just their onsets and that older adults require a greater word onset duration for recognition than young adults. In this study, young and older adults heard either just word onsets, word onsets followed by white noise indicating the full duration of the target word, or word onsets followed by a low-pass-filtered signal that indicated the number of syllables and syllabic stress (word prosody) in the absence of segmental information. Older adults required longer stimulus durations for word recognition under all conditions, with age differences in hearing sensitivity contributing significantly to this age difference. Within this difference, however, word recognition was facilitated by knowledge of word prosody to the same degree for young and older adults. These findings suggest, first, that listeners can detect and utilize word stress in making perceptual judgments and, second, that this ability remains spared in normal aging. PMID- 11386479 TI - Binaural speech recognition and the Stenger effect. AB - The present investigation examined the effects of systematically altering the balance between speech presentation levels to the 2 ears of 12 listeners with bilateral asymmetrical sensorineural hearing impairments. Speech-recognition scores for /VCV/ speech stimuli were obtained from each participant in quiet for 9 conditions ranging from monaural poorer ear only to monaural better ear only, with 7 intermediate conditions in which the sound balance between ears was varied in 5-dB steps. High-pass spectral shaping was provided to the poorer ear, and unshaped amplification was provided to the better ear. The results suggested that, as a group, varying the sound level in the better ear within -20 to +10 dB of the centered position did not significantly change the speech recognition for these participants. No evidence of binaural interference was obtained. Findings also showed that in binaural listening situations, the Stenger effect has little influence upon speech-recognition scores. Even when the listeners were unaware of speech being presented to the better ear, their speech-recognition score reflected the better ear's abilities. PMID- 11386480 TI - Modulations in respiratory and laryngeal activity associated with changes in vocal intensity during speech. AB - We tested the hypothesis that different strategies are used to alter tracheal pressure (Pt) during sustained and transient increases in intensity. It has been suggested that the respiratory system plays the primary role in Pt changes associated with alteration in overall intensity, whereas laryngeal adjustment is primary for transient change in Pt related to emphasis. Tracheal pressure, obtained via tracheal puncture, airflow (U), and laryngeal electromyography from the thyroarytenoid muscle (TA EMG) were collected from 6 subjects during sentence production at different intensity levels and with various stress patterns. Using a technique described in a previous study, we computed lower airway resistance (Rlaw) from measures of Pt and U obtained during a sudden change in upper airway resistance. We used this resistance value, together with direct measures of Pt and U during speech, to derive a time-varying measure of alveolar pressure (Pa), the pressure created by respiratory muscle activity and elastic recoil of the lungs. Pa provided a measure of respiratory drive that was unaffected by laryngeal activity. Laryngeal airway resistance (Rlx) and TA EMG provided measures of laryngeal activity. The results of this study indicated that, contrary to the outcome predicted by the hypothesis, there was no difference in the strategies used to alter Pt during sustained and transient increases in intensity. Although changes in both Pa and Rlx contributed to increase in Pt, the contribution of Pa was substantially greater. On average, Pa contributed to 94% and Rlx to 6% of the increase in Pt associated with vocal intensity. A secondary purpose of the study was to determine the extent to which laryngeal muscle activity was related to Rlx during speech. We found TA EMG activity increased with intensity but was not well correlated with Rlx, suggesting that when it contracts, the TA muscle may affect intensity by loosening the cover, which allows for greater amplitude of vocal fold vibration, without necessarily increasing laryngeal airway resistance. PMID- 11386481 TI - Individual and consensus judgments of disfluency types in the speech of persons who stutter. AB - Previous research has suggested that the reliability with which judges identify individual disfluency types, such as repetitions or prolongations of speech sounds, may be very poor. The use of disfluency types judgments in research and clinical applications is also complicated by important differences among the several disfluency-based characterizations of stuttered speech. In an attempt to address these problems, this study arranged for 30 judges to identify all disfluency types that they perceived to be present in 5-s audiovisually recorded speech stimuli, each in an Individual task and then with a partner in a Consensus task. Intrapair agreement and interpair agreement for occurrences of disfluency types (from Consensus conditions) were significantly higher than intrajudge and interjudge agreement for occurrences (from Individual conditions). Despite being higher than individual values, however, intrapair and interpair agreement for occurrences both averaged less than 50%. Results also showed that disfluency types judgments, interpreted in terms of three common disfluency-based definitions of stuttering, were not strongly related to previous assessments of whether these speech tokens contained or did not contain stuttering. When combined with previously available data, the present findings suggest caution in the use of disfluency types to describe or define stuttered speech. PMID- 11386482 TI - Acoustic duration changes associated with two types of treatment for children who stutter. AB - The purpose of this study was (a) to examine in young children the effects of Speech Motor Training (SMT) on selected temporal acoustic durations considered to be related to speech motor programming, (b) to compare the speech motor effects of that treatment with those of a treatment of childhood stuttering that did not directly incorporate speech motor control training (Extended Length of Utterance [ELU]), and (c) to examine the relation of acoustic duration changes to reduction of stuttering. Twelve children who stutter were recorded while repeating syllable sets /see text/ and /see text/ before and after SMT (n = 6) or ELU treatment (n = 6). Children who did not stutter served as matched reference groups. The syllables beginning with /p/ and /t/ were used as tokens for the acoustic measurement. Five measures served as indicators of temporal aspects of speech motor performance: vowel duration, stop gap duration, voice onset time, stop gap/vowel duration ratio, and total token duration. Results indicated that following SMT there was a significant increase in vowel duration and some reduction in stop gap duration that resulted in significantly reduced stop gap/vowel duration ratios. These acoustic effects were consistent across most participants. The ELU treatment reduced stuttering more than the SMT, but was not accompanied by significant effects on the selected temporal acoustic measures. These findings are compared with previous findings of increased vowel durations associated with fluency enhancement and stuttering treatment. We speculate that the increased vowel durations allow more time for speech motor planning and that stuttering is reduced moderately as a by-product of longer vowel durations. The mechanism(s) by which ELU treatment reduces stuttering did not appear to be captured by the dependent variables measured in this study. PMID- 11386483 TI - A comparison of equal-appearing interval scaling and direct magnitude estimation of nasal voice quality. AB - Listeners rated the nasality of synthesized vowels using two psychophysical scaling methods (equal-appearing interval scaling and direct magnitude estimation). A curvilinear relationship between equal-appearing interval ratings and direct magnitude estimations of nasality indicated that nasality is a prothetic rather than metathetic dimension. It also was shown that the use of direct magnitude estimation results in nasality ratings that are more consistent and reliable. The results of this experiment are discussed in relation to other studies that have examined the validity and reliability of equal-appearing interval scaling of voice quality. Additionally, there is a discussion of methodological issues for future research and the implications of the findings for clinical and research purposes. PMID- 11386484 TI - Multidimensional scaling of nasal voice quality. AB - Listeners judged the dissimilarity of pairs of synthesized nasal voices that varied on 3 dimensions. Separate nonmetric multidimensional scaling (MDS) solutions were calculated for each listener and the group. Similar 3-dimensional solutions were derived for the group and each of the listeners, with the group MDS solution accounting for 83% of the total variance in listeners' judgments. Dimension 1 ("Nasality") accounted for 54% of the variance, Dimension 2 ("Loudness") for 18% of the variance, and Dimension 3 ("Pitch") for 11% of the variance. The 3 dimensions were significantly and positively correlated with objective measures of nasalization, intensity, and fundamental frequency. The results of this experiment are discussed in relation to other MDS studies of voice perception, and there is a discussion of methodological issues for future research. PMID- 11386485 TI - A preliminary study of factors influencing perception of articulatory rate in Parkinson disease. AB - This investigation was a preliminary study of factors influencing perception of articulatory rate in idiopathic Parkinson disease (PD). Nine men with mild to moderate idiopathic PD and 9 healthy men of a similar age read the Farm Passage at habitual, fast, and slow reading rates. Spontaneous speech samples also were obtained. Speech severity for participants with PD was mild to moderate; prominent perceptual characteristics further suggested hypokinetic dysarthria. Nine listeners estimated the articulatory rate of speech runs (i.e., stretch of speech bounded by pauses) extracted from the reading and spontaneous speech samples. Regression analysis was used to explore the relationship between articulatory rate (i.e., syllables produced per second excluding pauses) and perceptual impressions of articulatory rate as well as the relationship between fundamental frequency (F0) range and perceptual estimates of articulatory rate. Regression functions predicting perceptual estimates of articulatory rate from F0 range were not significant for either speaker group. The regression analyses relating articulatory rate and perceptual estimates of articulatory rate, however, revealed a significant, positive relationship between the two measures for both speaker groups. There also was a nonsignificant trend for perception of articulatory rate to grow more rapidly for Parkinsonian speech samples. Although the current findings should be interpreted with caution until they have been validated by additional studies using much larger speaker populations and more extensive speech samples, the results hint at the possibility that perceptual impressions of articulatory rate in PD may overestimate the actual, physical rate. The results also highlight the importance of speech-language pathologists complimenting perceptual judgements of articulatory rate with physical measures. PMID- 11386486 TI - New technologies to improve clinical trials. PMID- 11386487 TI - The association between intravenous haloperidol and prolonged QT interval. AB - Although intravenous haloperidol (HAL) is an effective medication that is often prescribed to treat agitation, several instances of torsade de pointes or prolonged QT interval have been reported. To investigate the association between intravenous HAL and QT prolongation and between intravenous HAL and ventricular tachyarrhythmia, a cross-sectional cohort study was performed that included measuring corrected QT intervals (QTc) on an emergency basis before intravenous HAL and continuously monitoring electrocardiographic (ECG) findings after intravenous HAL. During a 2-month period, 47 patients received intravenous injections to control psychotic disruptive behavior. According to clinical practice, patients were divided as follows. The FZ-alone group was treated with intravenous flunitrazepam (FZ), and the FZ-plus-HAL group received intravenous FZ followed by intravenous HAL. Although the difference in the mean QTc immediately after intravenous FZ between the two groups was not significant, the mean QTc after 8 hours in the FZ-plus-HAL group was longer than that in the FZ-alone group (p < 0.001). Four patients in the FZ-plus-HAL group had a QTc of more than 500 msec after 8 hours. The change in QTc during 8 hours significantly differed between the two groups (t = 2.64, p > 0.05). Furthermore, the change in QTc was moderately correlated with the dose of intravenous HAL, as evidenced by a coefficient of correlation of 0.48 (p < 0.001). However, ventricular tachyarrhythmia was not detected among 307 patients within a 1-year period, although the ECG was continuously monitored for at least 8 hours after intravenous HAL. The modest nature of QTc prolongation and the apparent absence of ventricular tachyarrhythmia under continuous ECG monitoring indicate that QTc prolongation associated with intravenous HAL is not necessarily dangerous. However, in an emergency situation, clinicians cannot exclude patients predisposed to torsade de pointes, such as those with inherited ion channel disorders. Therefore, clinicians should be aware of the association between intravenous HAL and QT prolongation. PMID- 11386488 TI - The 35% CO2 hyperreactivity and clinical symptomatology in patients with panic disorder after 1 week of treatment with citalopram: an open study. AB - The effect of a short treatment (7 days) with citalopram on the reactivity to inhalations of 35% CO2 and 65% oxygen and on clinical symptomatology was investigated in 15 patients with panic disorder who had a positive response to 35% CO2 inhalation. An open study design was applied. On day 0, before starting drug treatment, and after 1 week of treatment, each patient underwent the 35% CO2 challenge, and clinical symptomatology was evaluated with psychometric scales. The results showed a significant reduction of CO2 reactivity and of scores on the anticipatory anxiety subscale of Panic Associated Symptoms Scale. These results confirm that the serotonergic system plays an important role in the modulation of CO2 hyperreactivity and suggest an early anxiolytic effect of citalopram in patients who have panic disorder and are hyperreactive to CO2. PMID- 11386489 TI - Pharmacokinetics and clinical effects of sublingual triazolam in pediatric dental patients. AB - The purpose of this investigation was to describe the pharmacokinetics of sublingual triazolam in children. Nine healthy children (64-98 months old) received 0.25 or 0.375 mg of sublingual triazolam before dental treatment. Plasma triazolam concentrations were measured by gas chromatography/mass spectrometry and analyzed by noncompartmental methods. The peak concentration was 4.9 +/- 2.0 ng/mL (mean +/- SD), time to peak was 75 +/- 32 minutes, the elimination half life was 91 +/- 32 minutes, and apparent clearance was 17.6 +/- 8.8 mL x kg(-1) x min(-1). Children were tested for gait ataxia, amnesia, and diplopia during a screening session and again after triazolam. Ninety minutes after drug administration, seven of nine children demonstrated ataxia, and three of nine demonstrated amnesia. Peak triazolam concentrations were similar in children with or without ataxia, but they were significantly higher in children with amnesia compared with those without amnesia. Six children demonstrated diplopia 30 and 120 minutes after triazolam; however, peak triazolam concentrations were similar in both groups. Sublingual administration was an acceptable alternative route of triazolam delivery in children. PMID- 11386490 TI - Reinforcing and subject-rated effects of methylphenidate and d-amphetamine in non drug-abusing humans. AB - The reinforcing effects of methylphenidate (20-40 mg), d-amphetamine (10-20 mg), and placebo were assessed in eight healthy, non-sleep-deprived, non-drug-abusing outpatient volunteers. A modified progressive-ratio schedule was used to assess drug reinforcement in which a sampling session always preceded a self administration session. During sampling sessions, volunteers received a drug dose to acquaint them with the drug effects. Drug doses were administered in eight identical capsules (i.e., each capsule contained 12.5% of the total dose). During self-administration sessions, which generally were conducted the next day, volunteers were given eight opportunities to work on a computer and could earn all, or some, of the capsules that were administered the previous day. To earn the first capsule, volunteers had to click a computer mouse 50 times. The number of clicks required to earn each additional capsule doubled (i.e., 100, 200, 400, 800, 1,600, 3,200, and 6,400 clicks). The dependent measure on this task was the break point (i.e., the last ratio completed). To characterize more fully the behavioral effects of methylphenidate and d-amphetamine, a battery of subject rated drug-effect questionnaires, performance tasks, and physiologic measures was also used. Both doses of d-amphetamine increased the break point significantly above placebo levels, whereas only the high dose of methylphenidate did so. Break point values for the doses of methylphenidate and d-amphetamine that maintained the greatest responding did not differ significantly. Methylphenidate and d amphetamine produced some stimulantlike subject-rated drug effects (e.g., increased ratings of "drug liking"). These data suggest that methylphenidate, like d-amphetamine, can function as a reinforcer under a modified progressive ratio schedule and, by inference, has at least some abuse potential in healthy, non-sleep-deprived, non-drug-abusing volunteers. PMID- 11386491 TI - Targeted use of naltrexone without prior detoxification in the treatment of alcohol dependence: a factorial double-blind, placebo-controlled trial. AB - Several studies have shown the opioid antagonist naltrexone to be effective when combined with psychosocial therapies for the treatment of patients who are dependent on alcohol with fixed medication and time (12 weeks). In this study, 121 nonabstinent outpatients with alcohol dependence (DSM-IV) were treated with sessions of cognitive coping skills (N = 67) or supportive therapy (N = 54) and either naltrexone 50 mg/day (N = 63) or placebo (N = 58) daily for the first 12 weeks and thereafter for 20 weeks only when craving alcohol (i.e., targeted medication) in a prospective one-center, dual, double-blind, randomized clinical trial. The dropout rate for all subjects was 16.5% during the first 12-week period and approximately twice that level by the end of the study. There were no significant group differences in study completion and therapy participation rates. After the continuous medication (12 weeks), the coping/naltrexone group had the best outcome, and coping/placebo had the worst. This difference remained during the targeted medication period (the following 20 weeks). Naltrexone was not better than placebo in the supportive groups, but it had a significant effect in the coping groups: 27% of the coping/naltrexone patients had no relapses to heavy drinking throughout the 32 weeks, compared with only 3% of the coping/placebo patients. The authors' data confirm the original finding of the efficacy of naltrexone in conjunction with coping skills therapy. In addition, their data show that detoxification is not required and that targeted medication taken only when craving occurs is effective in maintaining the reduction in heavy drinking. PMID- 11386492 TI - Antidepressants and ejaculation: a double-blind, randomized, placebo-controlled, fixed-dose study with paroxetine, sertraline, and nefazodone. AB - Antidepressant medication is often associated with sexual side effects. A double blind, placebo-controlled study in men with lifelong rapid ejaculation was performed to assess the effects of two selective serotonin (5-HT) reuptake inhibitors--paroxetine and sertraline--and the 5-HT2 antagonist and 5 HT/noradrenaline reuptake inhibitor nefazodone on the latency to ejaculate. Forty eight men with an intravaginal ejaculation latency time (IELT) of a maximum of 1 minute were randomly assigned to receive paroxetine (20 mg/day), sertraline (50 mg/day), nefazodone (400 mg/day), or placebo for 6 weeks. During the 1-month baseline and 6-week treatment period, IELTs were measured at home with a stopwatch. The trial was completed by 40 men. During the 6-week treatment period, the geometric mean IELT in the placebo group was stable at approximately 20 seconds. Analysis of variance revealed a between-group difference in the evolution of IELT delay over time (p = 0.002); the IELT after paroxetine and sertraline gradually increased to approximately 146 and 58 seconds, respectively, compared with 28 seconds in the nefazodone group. The paroxetine and sertraline groups differed significantly (p < 0.001 and p = 0.024, respectively) from placebo, but the nefazodone group did not (p = 0.85). Compared with baseline, paroxetine exerted the strongest delay in ejaculation, whereas sertraline delayed it only moderately. There was no clinically relevant delay in ejaculation with nefazodone. PMID- 11386493 TI - The relative efficacy of fluoxetine and manual-based self-help in the treatment of outpatients with bulimia nervosa. AB - A randomized, placebo-controlled study was conducted examining the singular and combined effects of fluoxetine and a self-help manual on suppressing bulimic behaviors in women with bulimia nervosa. A total of 91 adult women with bulimia nervosa were randomly assigned to one of four conditions: placebo only, fluoxetine only, placebo and a self-help manual, or fluoxetine and a self-help manual. Subjects were treated for 16 weeks. Primary outcome measures included self-reports of bulimic behaviors. Fluoxetine and a self-help manual were found to be effective in reducing the frequency of vomiting episodes and in improving the response rates for vomiting and binge-eating episodes. Furthermore, both factors were shown to be acting additively on the primary and secondary efficacy measures in this study. Results are discussed in relation to previous research and the implications for treatment of bulimia nervosa. PMID- 11386494 TI - Therapeutic drug monitoring of clozapine in relapse prevention: a five-year prospective study. AB - Twenty-three outpatients with schizophrenia (ICD-10 F20.xx) treated with clozapine (CZ) as monotherapy entered a prospective study on relapse prevention. Every 4 weeks, psychopathology was assessed by the Brief Psychiatric Rating Scale (BPRS), and plasma CZ and norclozapine levels were measured. Patients were enrolled after complete remission of positive symptoms for at least 4 months according to the psychosis cluster of the BPRS and at a mean of 3.3 years after their last hospitalization. At the time of enrollment, the median BPRS total score was 29 points (range, 19-48). Within 4 months, the baseline CZ plasma level was established as the mean of CZ levels from at least four subsequent measurements. These baseline plasma levels were considered as the optimal relapse preventing plasma CZ levels in the individual patients. When the patients were enrolled, they were considered to be prone to relapse. Relapse was defined as clinical deterioration, hospitalization, or both. Plasma levels were considered a prognostic factor, and patients were defined as at increased risk if plasma levels decreased by more than 40% from baseline CZ plasma level. The effect of plasma CZ levels on clinical outcome was evaluated by a Cox regression with plasma level as a time-dependent covariate. Within 46 months of enrollment, 32 episodes of relapse events in 10 patients were available for evaluation. Seventeen patients had a plasma level decrease of more than 40% at some point. In 12 of these, the decrease was present for more than 12% of the observation period. Eight patients of this group relapsed, and three of these had to be rehospitalized. Two patients relapsed, although their plasma levels decreased by more than 40% for less than 12% of the observation period. Within the first 2 years, relapse-free survival curves illustrate that both groups (episodes under elevated risk and episodes not under elevated risk) had identical relapse patterns, but from then on the relapse risk increased rapidly in the group with longer exposure to elevated risk. In a Cox model with a 40% decrease of plasma CZ levels as a dichotomous time-varying explanatory covariate, the risk ratio is 6 (95% confidence interval = 2-19, p = 0.003). The 10 patients who relapsed exhibited safe plasma levels (less than a 40% decrease from their baseline levels) for only 210 months, and 13 nonrelapsing patients had plasma levels defined as safe for 426 months. PMID- 11386495 TI - Psychological and pharmacological treatments of social phobia: a meta-analysis. AB - A meta-analysis of psychological and pharmacological treatments for social phobia was conducted to evaluate whether the various treatments differ in their efficacy for treating social phobia, whether they are more effective than wait-list and placebo controls, whether rates of attrition differ, and whether treatment gains are maintained at follow-up. A total of 108 treatment-outcome trials for social phobia met inclusion/exclusion criteria for the meta-analysis. Eleven treatment conditions were compared: wait-list control, pill placebo, benzodiazepines (BDZs), selective serotonin reuptake inhibitors (SSRIs), monoamine oxidase inhibitors, attention placebo, exposure (EXP), cognitive restructuring (CR), EXP plus CR, social skills training, and applied relaxation. The most consistently effective treatments for social phobia were pharmacotherapies. BDZs and SSRIs were equally effective and more effective than control conditions. Dropout rates were similar among all the active treatment conditions. Assessment of the durability of treatment gains for pharmacotherapies was not possible because an insufficient number of drug studies included follow-up data. The treatment gains of psychological therapies, although moderate, continued during the follow-up period. BDZs and SSRIs seem to be effective treatments for social phobia, at least in the short term. Recommendations for future research include assessing the long-term outcome for pharmacotherapies and evaluating the inclusion of a cognitive-behavioral treatment during the drug tapering period. PMID- 11386496 TI - Bupropion sustained-release for the treatment of dysthymic disorder: an open label study. AB - Many studies of antidepressants in the treatment of dysthymic disorder (DD) have been conducted, but none has included bupropion sustained-release (SR). The aim of this study was to provide preliminary data on the tolerability and effectiveness of bupropion SR for patients with DD. Twenty-one adult subjects meeting DSM-IV criteria for DD were enrolled in this 8-week open-label study. Bupropion SR was initiated at 150 mg/day and was increased to a maximum of 200 mg, twice daily. Response was defined as a 50% or greater decrease in score on the Hamilton Rating Scale for Depression (HAM-D). Of these 21 subjects, 15 (71.4%) responded to treatment. All paired sample t-tests were highly significant, demonstrating average improvement on all measures of symptomatology and functioning. Subject scores on the HAM-D decreased from 21.7 +/- 5.6 at baseline to 5.9 +/- 3.6 at week 8 (t[19] = 12.74, p < 0.001). The average final dosage was 364 mg/day. None of the subjects dropped out during the trial. Patients with a history of alcohol or chemical abuse were significantly less likely to respond to bupropion. Side effects were reported by eight subjects (38.1%), and the most frequently reported effects were headache, decreased appetite, insomnia, gastrointestinal problems, restlessness, and tremulousness. These findings suggest the effectiveness and high tolerability of bupropion SR for the treatment of DD. Double-blind prospective studies are needed for the comparison of bupropion SR to both placebo and other medications, assessing both initial and sustained responses to treatment. PMID- 11386497 TI - Concentrations of the enantiomers of fluoxetine and norfluoxetine after multiple doses of fluoxetine in cytochrome P4502D6 poor and extensive metabolizers. AB - Plasma concentrations of the enantiomers of fluoxetine (FLX) and norfluoxetine (NFLX) were measured at days 7, 14, and 23 of oral administration of 20 mg of racemic fluoxetine in 11 patients who were comedicated with risperidone. Eight patients were genotyped as being cytochrome P4502D6 extensive metabolizers (EMs) and three as cytochrome P4502D6 poor metabolizers (PMs). No statistically significant differences were calculated between EMs and PMs in the concentrations of (R)-FLX and (R)-NFLX for all days examined (day 23, mean +/- SD for (R)-FLX and (R)-NFLX in EMs, 16 +/- 5 and 29 +/- 20 ng/mL, respectively; in PMs, 16 +/- 1 and 20 +/- 2 ng/mL, respectively). However, concentrations of (S)-FLX and (S) NFLX were higher and lower, respectively, in PMs as compared with EMs (day 7, p = 0.037 and p = 0.036; day 14, p = 0.014 and p = 0.014; day 23, p = 0.068 and p = 0.038). On day 23, mean (S)-FLX and (S)-NFLX in EMs were (mean +/- SD) 39 +/- 26 and 63 +/- 26 ng/mL, and in PMs they were 88 +/- 7 and 19 +/- 2 ng/mL. This study confirms the results of the single-dose studies showing that CYP2D6 is involved in the demethylation of FLX to NFLX, with a stereoselectivity toward the (S) enantiomer. The data also clearly show that the CYP2D6 genotype has an important influence on the concentrations of the (S)- but not of the (R)-enantiomer of FLX and NFLX after multiple doses. PMID- 11386498 TI - Double-blind, controlled, crossover trial of inositol versus fluvoxamine for the treatment of panic disorder. AB - Only 70% of patients respond to current treatments for panic disorder, and many discontinue drugs because of side effects. myo-Inositol, a natural isomer of glucose and a precursor for the second-messenger phosphatidyl-inositol system, has previously been found superior to placebo in the treatment of depression, panic disorder, and obsessive-compulsive disorder (OCD), but a direct comparison with an established drug has never been performed. A double-blind, controlled, random-order crossover study was undertaken to compare the effect of inositol with that of fluvoxamine in panic disorder. Twenty patients completed 1 month of inositol up to 18 g/day and 1 month of fluvoxamine up to 150 mg/day. Improvements on Hamilton Rating Scale for Anxiety scores, agoraphobia scores, and Clinical Global Impressions Scale scores were similar for both treatments. In the first month, inositol reduced the number of panic attacks per week (mean and SD) by 4.0 (2) compared with a reduction of 2.4 (2) with fluvoxamine (p = 0.049). Nausea and tiredness were more common with fluvoxamine (p = 0.02 and p = 0.01, respectively). Because inositol is a natural compound with few known side effects, it is attractive to patients who are ambivalent about taking psychiatric medication. Continuing reports of inositol's efficacy in the treatment of depression, panic disorder, and OCD should stimulate replication studies. PMID- 11386499 TI - A pilot study of topiramate as monotherapy in the treatment of acute mania. AB - This small-scale pilot study was performed to grossly document safety and any evidence of efficacy of topiramate in bipolar disorder. Ten patients hospitalized for acute mania were given open-label topiramate monotherapy for up to 28 days. The mean Young Mania Rating Scale (YMRS) score decreased from 32 (range, 26-40) at baseline to 22 (range, 2-40) at the end of the study. Five patients exhibited evidence of moderate to marked improvement, three subjects had at least a 50% reduction in YMRS scores, and the other two patients experienced an improvement of 25% to 49% on the YMRS. The preliminary findings of this small series suggest that topiramate may be effective in acute mania. Double-blind controlled trials are now needed to further investigate the efficacy and safety of topiramate in bipolar disorder. PMID- 11386500 TI - Improvement of hallucinogen persisting perception disorder by treatment with a combination of fluoxetine and olanzapine: case report. PMID- 11386501 TI - Comments on "Serotonin syndrome during treatment with paroxetine and risperidone". PMID- 11386502 TI - Comments on "Hypotension associated with intravenous haloperidol and imipenem. PMID- 11386503 TI - Tiagabine, a specific gamma-aminobutyric acid transporter-1 inhibitor, in the treatment of psychosis. PMID- 11386504 TI - Exogenous melatonin, jet lag, and psychosis: preliminary case results. PMID- 11386505 TI - Hyperglycemia in patients with schizophrenia who are treated with olanzapine. PMID- 11386506 TI - Antiaggressive action of atypical antipsychotics in patients with schizophrenia. PMID- 11386507 TI - Treating tardive dyskinesia with ondansetron. PMID- 11386508 TI - Incidence and clinical significance of inducible atrial tachycardia in patients with atrioventricular nodal reentrant tachycardia. AB - INTRODUCTION: The purpose of this prospective study was to determine the prevalence and clinical significance of inducible atrial tachycardia in patients undergoing slow pathway ablation for AV nodal reentrant tachycardia who did not have clinically documented episodes of atrial tachycardia. METHODS AND RESULTS: Twenty-seven (15%) of 176 consecutive patients who underwent slow pathway ablation for AV nodal reentrant tachycardia were found to have inducible atrial tachycardia with a mean cycle length of 351+/-95 msec. The atrial tachycardia was sustained in 7 (26%) of 27 patients and was isoproterenol dependent in 20 patients (74%). The atrial tachycardia was not ablated or treated with medications, and the patients were followed for 9.7+/-5.8 months. Six (22%) of the 27 patients experienced recurrent palpitations during follow-up. In one patient each, the palpitations were found to be due to sustained atrial tachycardia, nonsustained atrial tachycardia, recurrence of AV nodal reentrant tachycardia, paroxysmal atrial fibrillation, sinus tachycardia, and frequent atrial premature depolarizations. Thus, only 2 (7%) of 27 patients with inducible atrial tachycardia later developed symptoms attributable to atrial tachycardia. CONCLUSION: Atrial tachycardia may be induced by atrial pacing in 15% of patients with AV nodal reentrant tachycardia. Because the vast majority of patients do not experience symptomatic atrial tachycardia during follow-up, treatment for atrial tachycardia should be deferred and limited to the occasional patient who later develops symptomatic atrial tachycardia. PMID- 11386510 TI - Electrospatial mapping technologies: here to stay or gone tomorrow? PMID- 11386509 TI - Clinical usefulness of a multielectrode basket catheter for idiopathic ventricular tachycardia originating from right ventricular outflow tract. AB - INTRODUCTION: It often is difficult to determine the optimal ablation site for idiopathic ventricular tachycardia (VT) originating from the right ventricular outflow tract (RVOT) when the VT or premature ventricular complex (PVC) does not occur frequently. The aim of our study was to evaluate the usefulness of a multielectrode basket catheter for ablation of idiopathic VT originating from the RVOT. METHODS AND RESULTS: Radiofrequency (RF) catheter ablation was performed using a 4-mm tip, quadripolar catheter in 50 consecutive patients with 81 VTs originating from the RVOT with (basket group = 25 patients with 45 VTs) or without (control group = 25 patients with 36 VTs) predeployment of a multielectrode basket catheter composed of 64 electrodes. Deployment of the multielectrode basket catheter was possible and safe in all 25 patients in the basket group. Ablation was successful in 25 (100%) of 25 patients in the basket group and in 22 (88%) of 25 patients in the control group. The total number of RF applications and the number of RF applications per PVC morphology did not differ between the two groups. However, both the fluoroscopic and ablation procedure times per PVC morphology were shorter in the basket group than in the control group (36.8+/-14.1 min vs 52.0+/-32.5 min, P = 0.04; 60.0+/-14.6 vs 81.5+/-51.2 min, P = 0.05). This difference was more pronounced in the 29 patients in whom VT or PVC was not frequently observed. CONCLUSION: The multielectrode basket catheter is safe and useful for determining the optimal ablation site in patients with idiopathic VT originating from the RVOT, especially in those without frequent VT or PVC. PMID- 11386511 TI - Worldwide clinical experience with a new dual-chamber implantable cardioverter defibrillator system. AB - INTRODUCTION: Management of atrial tachyarrhythmias represents a significant challenge in patients with implantable cardioverter defibrillators (ICDs). Drug therapy of these arrhythmias is limited by moderate efficacy, ventricular proarrhythmia, and drug-device interactions. This study tested the safety and efficacy of a new dual-chamber ICD to detect and treat atrial as well as ventricular tachyarrhythmias. METHODS AND RESULTS: A dual-chamber ICD (Medtronic 7250 Jewel AF) was implanted in 293 of 303 patients at 49 centers in Europe, Canada, and North America. Specific data were collected at implant and during a mean follow-up period of 7.9+/-4.7 months. There were no clinically evident failures to detect and treat ventricular arrhythmias. In patients with at least one of the dual-chamber detection criteria activated, 1,056 of 1,192 episodes of ventricular tachycardia or fibrillation detected were judged to be appropriate (89% positive predictive accuracy). Therapy efficacy was 100% in the ventricular fibrillation zone and 98% in the ventricular tachycardia zone. Positive predictive accuracy for detection of atrial episodes was 95% (1,052/1,107). For episodes classified as atrial tachycardia by the device, the efficacy of atrial antitachycardia pacing and high-frequency (50-Hz) burst pacing was 55% and 17%, respectively. High-frequency burst pacing terminated 16.8% of episodes classified as atrial fibrillation, and atrial defibrillation had an estimated efficacy of 76%. The actuarial estimates of 6-month complication-free survival and total survival were 88% and 94%, respectively. CONCLUSION: This novel dual-chamber ICD is capable of safely and effectively discriminating atrial from ventricular tachyarrhythmias and of treating atrial tachyarrhythmias without compromising detection and treatment of ventricular tachyarrhythmias. PMID- 11386512 TI - Radiofrequency catheter ablation of idiopathic left ventricular outflow tract tachycardia: utility of intracardiac echocardiography. AB - INTRODUCTION: The site of origin of idiopathic ventricular tachycardia (VT) arising from the left ventricular outflow tract (LVOT) may be closely related to the aortic valve leaflets, and radiofrequency (RF) delivery potentially can damage them. Intracardiac echocardiography (ICE) can identify accurately the ablation electrode and anatomic landmarks, and contact with the endocardium can be easily assessed. The aim of this study was to define the utility and the accuracy of ICE in guiding RF ablation of idiopathic VT of the LVOT. METHODS AND RESULTS: Five consecutive patients (all men; mean age 20.4 years, range 16 to 25) symptomatic for idiopathic VT underwent RF ablation. A 9-French, in-sheath catheter with a 9-MHz ultrasound transducer was inserted through the femoral vein and positioned in the His-bundle region or right ventricular outflow tract to provide a clear view of the aortic root. Local earliest ventricular activation during tachycardia and pace mapping were used to identify the ablation site. Idiopathic VT was ablated successfully in all patients using a median of two RF pulses, delivered during tachycardia. High-resolution images of the aortic valve and ablation electrode were achievable in all cases. Direct vision of ablation electrode-endocardial contact in the outflow tract was assessed easily in all patients. CONCLUSION: Idiopathic VT of the LVOT can be treated successfully with RF ablation. ICE can accurately guide catheter ablation and identify anatomic landmarks, endocardial contact, and ablation electrode movement. PMID- 11386513 TI - Idiopathic ventricular outflow tract tachycardia: where does it originate? PMID- 11386514 TI - Female gender is a risk factor for drug-induced long QT and cardiac arrhythmias in an in vivo rabbit model. AB - INTRODUCTION: Clinical observations and in vitro experimental data indicate that females have a longer QT interval than males, which is associated with a higher risk of drug-induced cardiac arrhythmias. Little is known about this gender difference in anesthetized animals, which may affect the outcome of in vivo drug tests. METHODS AND RESULTS: We evaluated potential gender differences in ventricular repolarization (QT, QTc, JT, and JTc interval) and its dispersion, as well as in its response to dofetilide, an IKr blocker, in anesthetized rabbits challenged with the alpha1-adrenoceptor agonist methoxamine. A 12-lead ECG was recorded during the experiments. At baseline, there were no significant gender differences in ventricular repolarization values in male and female rabbits under anesthesia. Dofetilide (0.04 mg/kg/min IV for 60 min; n = 10 per gender) produced marked prolongation of the ventricular repolarization time and its dispersion, associated with a high incidence of polymorphic ventricular tachycardia (PVT; 100% in females vs 80% in males) and ventricular fibrillation (VF; 80% in females vs 50% in males; P > 0.05). QT and JT interval at 2 minutes as well as QT and JT dispersion at 10 and 30 minutes during dofetilide infusion were significantly higher in female than in male rabbits. After 30 minutes of dofetilide infusion, 10 of 10 female rabbits had severe cardiac arrhythmias (complete AV block, PVT, or VF), so ECG parameters were impossible to assess (vs 3/10 males with severe cardiac arrhythmias; P < 0.05). During dofetilide infusion, female rabbits developed complete AV block, PVT, or VF at doses about 50% lower than those given to males. CONCLUSION: The present study indicates that female rabbits are more susceptible to drug-induced long QT and cardiac arrhythmias than are male rabbits; therefore, female rabbits are more appropriate for testing drug-induced cardiac arrhythmias. PMID- 11386515 TI - Prevention of drug-induced long QT syndrome: gender, chemistry, and education. PMID- 11386516 TI - Method for simultaneous epicardial and endocardial mapping of in vivo canine heart: application to atrial conduction properties and arrhythmia mechanisms. AB - INTRODUCTION: It has been suggested that the three-dimensional structure of the atria may be crucial in arrhythmogenesis; however, previous in vivo atrial activation mapping studies have been limited to either endocardial or epicardial approaches. METHODS AND RESULTS: To investigate the role of endocardial and epicardial structures and their interaction in atrial conduction and arrhythmias, we used five epicardial plaques and two intra-atrial balloon arrays to record a total of 368 unipolar electrograms from the entire epicardial and endocardial surface of both atria. During regular 1:1 pacing from the right atrial appendage, right atrial endocardial activation spread considerably faster than epicardial (total activation time 45+/-12 msec vs 60+/-19 msec, respectively [mean +/- SD]; P < 0.05), pointing to preferential conduction over structures like the crista terminalis and pectinate muscles. No such differences were noted in the left atrium. Transseptal spread occurred via discrete anterior and posterior pathways, causing separate breakthroughs in anterior and posterior atrial regions, respectively. Dissociation between septal pathways played a role in reentry during vagal atrial fibrillation. In 2 of 4 dogs with atrial fibrillation associated with congestive heart failure, single macroreentrant circuits involving endocardial and epicardial components were revealed during the arrhythmia. CONCLUSION: We conclude that activation mapping using simultaneous recording from both epicardial and endocardial surfaces provides potentially important insights into the mechanisms of atrial conduction and arrhythmogenesis. PMID- 11386517 TI - Critically timed auxiliary shock to weak field area lowers defibrillation threshold. AB - INTRODUCTION: This study tested the hypothesis that the defibrillation threshold (DFT) can be lowered by delivering a weak auxiliary shock in conjunction with a stronger primary shock to the cardiac region where the primary shock electric field is weakest. METHODS AND RESULTS: Eight swine were studied in each of two study parts. In both parts, DFTs were determined for dual shocks delivered through two electrode pairs. The biphasic primary shock was delivered through electrodes in the right ventricle and superior vena cava. The auxiliary shock was delivered through a separate electrode in the superior vena cava and a left ventricular electrode placed where the primary shock field was presumed to be weakest. In part I, a monophasic auxiliary shock of 50, 100, or 150 V was delivered either simultaneously with or 1, 20, or 40 msec before primary shock. When auxiliary shock was delivered simultaneously with or 1 msec before primary shock, DFT energy was reduced by approximately 50% compared with primary shock alone. In part II, a 150-V monophasic or biphasic auxiliary shock of either polarity was delivered 1 msec before or after primary shock. Regardless of waveform or polarity, all auxiliary shock delivered before primary shock lowered DFT energy by approximately 30% compared with primary shock alone. Depending on waveform and polarity, auxiliary shock delivered after primary shock either did not significantly change the DFT or elevated the DFT compared with primary shock alone. CONCLUSION: Application of a small auxiliary shock, just before or simultaneously with a primary shock, to the cardiac region where the primary shock field is weakest significantly lowers DFT. PMID- 11386518 TI - Role of calcium in acute hyperthermic myocardial injury. AB - INTRODUCTION: We hypothesized that intracellular calcium overload may play an important role in heat-induced myocardial injury. This postulate was investigated using a model of isolated guinea pig papillary muscle in which resting tension was measured as an indirect indicator of cytosolic free-calcium concentration and the fluorescence changes of Fluo-3 AM dye was measured as a direct indicator of cytosolic free-calcium concentration. METHODS AND RESULTS: Excised guinea pig right ventricular papillary muscles were attached to a force transducer in a high flow tissue bath and superfused with Tyrode's solution at 37 degrees +/- 0.5 degrees C. The temperature was rapidly changed to between 38.0 degrees and 56.0 degrees C for 60 seconds and then returned to 37.0 degrees C. Hyperthermia caused a reversible increase in resting tension at temperatures between 45 degrees and 50 degrees C and irreversible contracture at > or =50 degrees C. Rapid cooling contracture experiments and experiments measuring fluorescence of myocytes loaded with 5 microM Fluo-3 AM dye demonstrated that the hyperthermia-induced rise in resting tension was likely due to an increase in intracellular calcium content. Inhibition of the sarcoplasmic reticulum calcium pump with 20 microM thapsigargin resulted in irreversible contracture of the papillary muscles at temperatures between 45 degrees and 50 degrees C and significant increases in Fluo-3 fluorescence at 48 degrees C. Blockade of sarcolemmal calcium channels with 0.5 mM cadmium or 40 microM verapamil did not attenuate the heat-induced increase in resting tension and Fluo-3 fluorescence. CONCLUSION: Hyperthermia causes an increase in resting tension of cardiac muscle that most likely is mediated by a calcium channel-independent increase in calcium permeability of the sarcolemmal membrane and/or release of stored intracellular calcium. PMID- 11386519 TI - Altering ventricular activation remodels gap junction distribution in canine heart. AB - INTRODUCTION: Prolonged arrhythmic or paced ventricular activation causes persistent changes in myocardial conduction and repolarization that may result from altered electrotonic current flow, for which gap junctional coupling is the principal determinant. Remodeling of gap junctions and their constituent connexins modifies conduction and has been causally implicated in reentrant arrhythmogenesis. We hypothesized conversely that altering the pattern of ventricular activation causes gap junctional remodeling. METHODS AND RESULTS: Seven dogs were paced from the left ventricular (LV) epicardium (VVO, approximately 120 beats/min) for 21 days before excision of transmural LV samples that were divided into endomyocardial, mid-myocardial, and epimyocardial layers. Another five paced dogs had recording electrodes attached to multiple LV sites. All 12 dogs developed characteristic pacing-induced persistent T wave changes of cardiac memory. After 21 days of pacing, the ventricularly paced QRS duration prolonged by a mean of 4 msec over baseline (P < 0.05), a change that was associated with significant slowing of intraventricular conduction to local sites. These changes in QRS duration and repolarization were associated with a reduction in epimyocardial connexin43 expression on quantitative Western blotting of LV myocardium from close to, but not distant from, the pacing site (61.7+/ 18.4 vs 100.9+/-34.0; P < 0.02) and a marked disruption in immunolabeled connexin43 distribution in epimyocardium only. CONCLUSION: Spatially distinct transmural and regional gap junctional remodeling is a consequence of abnormal ventricular activation and is associated with consistent changes in activation that may alter patterns of repolarization and facilitate reentrant arrhythmogenesis. PMID- 11386520 TI - Memory gaps in the heart. PMID- 11386521 TI - Reentry site during fibrillation induction in relation to defibrillation efficacy for different shock waveforms. AB - INTRODUCTION: Unsuccessful defibrillation shocks may reinitiate fibrillation by causing postshock reentry. METHODS AND RESULTS: To better understand why some waveforms are more efficacious for defibrillation, reentry was induced in six dogs with 1-, 2-, 4-, 8-, and 16-msec monophasic and 1/1- (both phases 1 msec) 2/2-, 4/4-, and 8/8-msec biphasic shocks. Reentry was initiated by 141+/-15 V shocks delivered from a defibrillator with a 150-microF capacitance during the vulnerable period of paced rhythm (183+/-12 msec after the last pacing stimulus). The shock potential gradient field was orthogonal to the dispersion of refractoriness. Activation was mapped with 121 electrodes covering 4 x 4 cm of the right ventricular epicardium, and potential gradient and degree of recovery of excitability were estimated at the sites of reentry. Defibrillation thresholds (DFTs) were estimated by an up-down protocol for the same nine waveforms in eight dogs internally and in nine other dogs externally. DFT voltages for the different waveforms were positively correlated with the magnitude of shock potential gradient and negatively correlated with the recovery interval at the site at which reentry was induced by the waveform during paced rhythm for both internal (DFT = 1719 + 64.5VV - 11.1RI; R2 = 0.93) and external defibrillation (DFT = 3445 + 150VV - 22RI; R2 = 0.93). CONCLUSION: The defibrillation waveforms with the lowest DFTs were those that induced reentry at sites of low shock potential gradient, indicating efficacious stimulation of myocardium. Additionally, the site of reentry induced by waveforms with the lowest DFTs was in myocardium that was more highly recovered just before the shock, perhaps because this high degree of recovery seldom occurs during defibrillation due to the rapid activation rate during fibrillation. PMID- 11386523 TI - Toward a mechanism-based understanding of atrial fibrillation. PMID- 11386522 TI - Focal atrial fibrillation: experimental evidence for a pathophysiologic role of the autonomic nervous system. AB - INTRODUCTION: Focal paroxysmal atrial fibrillation (AF) was shown recently to originate in the pulmonary veins (PVs) and superior vena cava (SVC). In the present study, we describe an animal model in which local high-frequency electrical stimulation produces focal atrial activation and AF/AT (atrial tachycardia) with electrogram characteristics consistent with clinical reports. METHODS AND RESULTS: In 21 mongrel dogs, local high-frequency electrical stimulation was performed by delivering trains of electrical stimuli (200 Hz, impulse duration 0.1 msec) to the PVs/SVC during atrial refractoriness. Atrial premature depolarizations (APDs), AT, and AF occurred with increasing high frequency electrical stimulation voltage. APD/AT/AF originated adjacent to the site of high-frequency electrical stimulation and were inducible in 12 of 12 dogs in the SVC and in 8 of 9 dogs in the left superior PV (left inferior PV: 7/8, right superior PV: 6/8; right inferior PV: 4/8). In the PVs, APDs occurred at 13+/-8 V and AT/AF at 15+/-9 V (P < 0.01; n = 25). In the SVC, APDs were elicited at 19+/-6 V and AT/AF at 26+/-6 V (P < 0.01; n = 12). High-frequency electrical stimulation led to local refractory period shortening in the PVs. The response to high-frequency electrical stimulation was blunted or prevented after beta receptor blockade and abolished by atropine. In vitro, high-frequency electrical stimulation induced a heterogeneous response, with shortening of the action potential in some cells (from 89+/-35 msec to 60+/-22 msec; P < 0.001; n = 7) but lengthening of the action potential and development of early afterdepolar izations that triggered APD/AT in other cells. Action potential shortening was abolished by atropine. CONCLUSION: High-frequency electrical stimulation evokes rapid ectopic beats from the PV/SVC, which show variable degrees of conduction block to the atria and induce AF, resembling findings in patients with focal idiopathic paroxysmal AF. The occurrence of the arrhythmia in this animal model was likely due to alterations in local autonomic tone by high-frequency electrical stimulation. Further research is needed to prove absolutely that the observed effects of high-frequency electrical stimulation were caused by autonomic nerve stimulation. PMID- 11386524 TI - Atypical left atrial flutter after intraoperative radiofrequency ablation of chronic atrial fibrillation: successful ablation using three-dimensional electroanatomic mapping. AB - Curative treatment of chronic atrial fibrillation (AF) remains a challenging task for electrophysiologists. Eliminating the initiating triggers by focal radiofrequency ablation in a subset of patients with paroxysmal AF and modifying the maintaining substrate by performing linear lesions within the left atrium in patients with prolonged episodes of AF are among the alternative approaches for management of these patients. Recently, a new intraoperative treatment procedure aimed at eliminating left atrial anatomic "anchor" reentrant circuits by induction of contiguous lesions using radiofrequency energy under direct vision was introduced. However, atypical left atrial flutter may occur during follow-up after intraoperative ablation of AF. These arrhythmias most likely are due to discontinuities in linear lesions; therefore, they can be successfully mapped and ablated in a subsequent percutaneous catheter ablation procedure. We report and discuss the case of a patient who underwent successful intraoperative ablation of chronic AF, but who developed atypical left atrial flutter postoperatively. Three dimensional nonfluoroscopic electroanatomic mapping revealed a gap in the linear lesion line connecting the left upper and right upper pulmonary vein orifices. Ablation at the exit site of the breakthrough was successful. PMID- 11386525 TI - Supraventricular tachycardia-ventricular tachycardia discrimination algorithms in implantable cardioverter defibrillators: state-of-the-art review. AB - To reduce inappropriate therapy of supraventricular tachycardia (SVT), implantable cardioverter defibrillators (ICDs) include algorithms to discriminate ventricular tachycardia (VT) from SVT. Dual-chamber algorithms analyze atrial and ventricular rates or AV relationship. They provide advantages over single-chamber algorithms, but introduce new ways to detect SVT as VT inappropriately and to underdetect VT. Unlike pacemakers, dual-chamber ICDs require accurate atrial sensing during high ventricular rates. A postventricular atrial blanking period prevents oversensing of far-field R waves as atrial electrograms, but causes underdetection of atrial fibrillation during high ventricular rates. Tachycardias with 1:1 AV relationship and VT during atrial tachyarrhythmias present specific SVT-VT discrimination problems. The first dual-chamber algorithms performed comparably to single-chamber algorithms. Present dual-chamber algorithms correct some limitations of earlier versions. PMID- 11386526 TI - Puzzling gender repolarization gap. PMID- 11386527 TI - Abrupt slowing of an escape pacemaker: what is the mechanism? PMID- 11386528 TI - Atrioventricular junction ablation performed via a patent foramen ovale. PMID- 11386529 TI - A worrisome experience during testing of an implantable cardioverter defibrillator (ICD). PMID- 11386530 TI - A worrisome experience during testing of an implantable cardioverter defibrillator (ICD). PMID- 11386531 TI - Transient complete AV block during slow pathway radiofrequency ablation, with continuation of the atrial tachycardia. PMID- 11386532 TI - Investigating trends in acoustics research from 1970-1999. AB - Text data mining is a burgeoning field in which new information is extracted from existing text databases. Computational methods are used to compare relationships between database elements to yield new information about the existing data. Text data mining software was used to determine research trends in acoustics for the years 1970, 1980, 1990, and 1999. Trends were indicated by the number of published articles in the categories of acoustics using the Journal of the Acoustical Society of America (JASA) as the article source. Research was classified using a method based on the Physics and Astronomy Classification Scheme (PACS). Research was further subdivided into world regions, including North and South America, Eastern and Western Europe, Asia, Africa, Middle East, and Australia/New Zealand. In order to gauge the use of JASA as an indicator of international acoustics research, three subjects, underwater sound, nonlinear acoustics, and bioacoustics, were further tracked in 1999, using all journals in the INSPEC database. Research trends indicated a shift in emphasis of certain areas, notably underwater sound, audition, and speech. JASA also showed steady growth, with increasing participation by non-US authors, from about 20% in 1970 to nearly 50% in 1999. PMID- 11386533 TI - Scattering of elastic waves in heterogeneous media with local isotropy. AB - The scattering of elastic waves in heterogeneous media is discussed. Explicit expressions are derived for the attenuation of longitudinal and transverse elastic waves in terms of the statistics of the density and Lame parameter fluctuations. The derivation is based upon diagrammatic methods with the problem posed in terms of the Dyson equation. The Dyson equation is solved for the mean field response. The results are given here in a straightforward manner, in which the attenuations reduce to simple integrals on the unit circle. The medium is assumed statistically homogeneous and statistically isotropic. This model, with assumed local isotropic properties, is expected to apply to many materials. PMID- 11386534 TI - Tomography of joint P-wave traveltime and polarization data: a simple approach for media with low to moderate velocity gradients. AB - An elastic wave tomography method utilizing joint traveltime and polarization data is proposed that is computationally simpler than the existing methods [Hu and Menke, Geophys. J. Int. 110, 63 (1992); Farra and Begat, Geophys. J. Int. 121, 371 (1995)]. In the linearization problem for the use of polarization data, we start with ray perturbation theory and assume that the medium is weakly inhomogeneous. Then the problem formulation for polarization data is approximately expressed as a linear integral of the gradient of the medium slowness perturbation along a reference ray. We call this a quasi-linear approximation which ignores the effect of the perturbation of the ray position on the first-order perturbation of the ray slowness vector. To efficiently obtain the solution for multi-data sets, a quadratic objective functional is constructed by including the data misfit terms and a model constraint term. Then a new conjugate gradient type of iterative reconstruction algorithm is developed to solve this minimization problem. This algorithm is also an extension of the conjugate gradient approach for standard least-squares problems. The feasibility and capability of the proposed tomography method is illustrated by conducting both noise-free and noisy synthetic experiments in a cross-hole geometry. The numerical results demonstrate that the additional use of polarization data not only improves the image quality, but also has a stabilizing effect on the iterative tomography solution. However, the limitation of the method is that it becomes inaccurate if the velocity variations in the medium change rapidly with position. PMID- 11386535 TI - Visualization of multiple incoherent sources by the backward prediction of near field acoustic holography. AB - When there are multiple noise sources which share the same frequency bands, a sound field measured or estimated by near-field acoustic holography (NAH) is obviously the combination of the sound fields generated by the multiple sources. The objective of this paper is to estimate the sound field generated by each source when the coherence functions between sources are zeros. This objective can be achieved by obtaining signals coherent to sources. This paper proposes a method to obtain the coherent signals by using the spatial information of NAH. The proposed method obtains the coherent signals from pressure signals at source positions estimated by the backward prediction of NAH. Thus it does not require any prior information on source positions, unlike the conventional method, which placed sensors close to sources in order to obtain the coherent signals. The proposed method was verified by a numerical simulation using two incoherent monopole sources. PMID- 11386536 TI - Adaptive time-reversal mirror. AB - The time-reversal mirror uses the received signal from a probe source to refocus the signal at the probe source location by backpropagating the time-reversed version of the received signal. In this study, an adaptive method is described to steer a null to an arbitrary position in a waveguide while maintaining a distortionless response at the probe source location. As an application, selective focusing in free space is demonstrated. PMID- 11386537 TI - Roughness characterization of porous soil with acoustic backscatter. AB - The use of acoustics to determine the pore properties of soils, such as porosity, permeability, and tortuosity, is well established. A theoretical surface impedance and complex bulk wavenumber was developed by K. Attenborough for porous media that incorporated the soil pore properties as parameters [J. Acoust. Soc. Am. 73, 785-799 (1983)]. Acoustic level difference measurements were used as a noninvasive means of finding the soil pore properties. Acoustic reflection measurements showed that the sound field over porous rough surfaces is modified by the surface impedance and by surface roughness. It is not possible to separate the signal modification due to impedance and the signal modification from roughness scattering in a forward scattering measurement. In order to accurately determine the soil pore properties, the roughness effects must be known independently from the surface impedance. A means of measuring roughness apart from impedance would allow the effects of roughness to be taken out of the level difference measurements. The underwater acoustics community has used acoustic backscatter for many years to examine surface roughness. The feasibility of adapting these acoustic backscatter techniques to outdoor porous soil surfaces is examined. PMID- 11386538 TI - Point load wave excitation in multi-layered solids: experiments and model verification. AB - Studies of transient wave propagation in a single plate, a two-layer plate structure, and a two-layer-on-a-half-space structure are reported. Experimental results are used to verify the accuracy of a new model for wave propagation in layered solids. The experiments are performed using point load excitation and point detection at the surface of the layered structure. The basis of the new wave propagation model is first introduced. Then, experiments performed on a single plate are described. Results from different wave sources, plate materials, and wave sensors are compared with each other and with computed results from the model. The transient source function for a pencil-lead-break wave source is obtained. Next, the results of experiments performed on a two-layer plate structure (aluminum bonded to stainless steel) are reported. The cases of stainless steel atop aluminum and aluminum atop stainless steel are considered. The effects of the second layer on the transient displacement at the top surface are discussed. Finally, a two-layer-on-a-half-space structure consisting of a stainless steel plate as the top layer, an aluminum plate as the second layer and a thick acrylic resin block as the half-space is studied. The obtained experimental transient measurements agree with the computed results for all cases, thus verifying the accuracy of the model. PMID- 11386539 TI - Time-frequency representations of Lamb waves. AB - The objective of this study is to establish the effectiveness of four different time-frequency representations (TFRs)--the reassigned spectrogram, the reassigned scalogram, the smoothed Wigner-Ville distribution, and the Hilbert spectrum--by comparing their ability to resolve the dispersion relationships for Lamb waves generated and detected with optical techniques. This paper illustrates the utility of using TFRs to quantitatively resolve changes in the frequency content of these nonstationary signals, as a function of time. While each technique has certain strengths and weaknesses, the reassigned spectrogram appears to be the best choice to characterize multimode Lamb waves. PMID- 11386540 TI - Exact solutions for transient spherical radiation. AB - Closed-form, analytic solutions are derived for exterior wave fields generated by the transient, axisymmetric motion of a spherical boundary. These are intended for use as benchmark problems for assessing the accuracy and correctness of transient numerical schemes. Classical Laplace transform methods are used. The derivation of these solutions is presented in sufficient detail to permit their reconstruction by the reader for multipoles of order n where 0 < or = n < or = 25. Composite solutions can also be obtained by superposition. A novel solution of this type is presented for the wave field generated by the transient motion of a spherical piston in a spherical baffle. The transient wave fields obtained in this way are shown to be consistent with steady time-harmonic solutions for large times, and with high-resolution transient numerical solutions at finite times. PMID- 11386541 TI - Energy losses in an acoustical resonator. AB - A one-dimensional model has recently been developed for the analysis of nonlinear standing waves in an acoustical resonator. This model is modified to include energy losses in the boundary layer along the resonator wall. An investigation of the influence of the boundary layer on the acoustical field in the resonator and on the energy dissipation in the resonator is conducted. The effect of the boundary layer is taken into account by introducing an additional term into the continuity equation to describe the flow from the boundary layer to the volume. A linear approximation is used in the development of the boundary layer model. In addition to the viscous attenuation in the boundary layer, the effect of acoustically generated turbulence is modeled by an eddy viscosity formulation. Calculatons of energy losses and a quality factor of a resonator are included into the numerical code. Results are presented for resonators of three different geometries: a cylinder, a horn cone, and a bulb-type resonator. A comparison of measured and predicted dissipation shows good agreement. PMID- 11386542 TI - Atmospheric scattering for varying degrees of saturation and turbulent intermittency. AB - Atmospheric turbulence is inherently inhomogeneous and intermittent. Short periods of high activity are embedded in longer periods of relative calm. Local spatial and temporal changes in sound speed associated with this intermittency increase the likelihood of measuring large values of scattered acoustic signals. Previous work successfully predicted the probability density functions (pdf's) of fully saturated, scattered signals measured within an acoustic shadow zone [Wilson et al., J. Acoust. Soc. Am. 99, 3393-3400 (1996)]. The more general case of incompletely saturated scattering is considered in this paper; using the Rice Nagakami distribution a theory is developed. The predicted intensity pdf has two free parameters: one to describe the degree of intermittency and a second for the degree of saturation. For validation purposes, outdoor propagation measurements were made over a flat, hard ground at ranges of 146-283 m and at frequencies of 50-540 Hz. The saturation parameter was determined from the acoustic data and also estimated from the turbulence conditions. The degree of saturation increased with frequency, and measured intensity pdf's were found to be in excellent agreement with the theory. PMID- 11386543 TI - Sound propagation in a turbulent atmosphere near the ground: an approach based on the spectral representation of refractive-index fluctuations. AB - A new, rigorous approach is presented for the computation of the fluctuating field of a monopole source in a nonrefracting, turbulent atmosphere above a ground surface. The time-averaged sound pressure level is considered, as well as statistical distributions of the sound pressure level. The computation is based on the Rytov solution of the wave equation for a turbulent medium, evaluated for the half-space above the ground surface. The solution takes into account the ground reflection of scattered waves, which has been neglected in previous work on this subject. The present approach is based on a Fourier-Stieltjes representation of refractive-index fluctuations, and makes use of a turbulent image atmosphere to account for the ground reflection of scattered waves. This approach is rigorous only for a rigid ground surface, but it is shown that it also yields a good approximation for a finite-impedance ground surface. The accuracy of the solution is demonstrated by comparison with results of numerical computations with the parabolic equation method for a turbulent atmosphere. The assumption of a nonrefracting atmosphere implies that direct application of the solution is limited to propagation over relatively small distances. However, this study can also be considered as a basis for a generalized solution for a downward refracting atmosphere, which can be applied for larger propagation distances. PMID- 11386544 TI - Sound propagation in a turbulent atmosphere near the ground: a parabolic equation approach. AB - The interference of the direct wave from the point source to the receiver and the wave reflected from the impedance ground in a turbulent atmosphere is studied. A parabolic equation approach for calculating the sound pressure p at the receiver is formulated. Then, the parabolic equation is solved by the Rytov method yielding expressions for the complex phases of direct and ground-reflected waves. Using these expressions, a formula for the mean squared sound pressure [absolute value(p)2] is derived for the case of anisotropic spectra of temperature and wind velocity fluctuations. This formula contains the "coherence factor," which characterizes the coherence between direct and ground-reflected waves. It is shown that the coherence factor is equal to the normalized coherence function of a spherical sound wave for line-of-sight propagation. For the case of isotropic turbulence, this result allows one to obtain analytical formulas for [absolute value(p)2] for the Kolmogorov, Gaussian, and von Karman spectra of temperature and wind velocity fluctuations. Using these formulas, the effects of temperature and wind velocity fluctuations, and the effects of different spectra of these fluctuations on the mean squared sound pressure, are numerically studied. Also the effect of turbulent anisotropy on the interference of direct and ground reflected waves is numerically studied. Finally, it is shown that the mean squared sound pressure [absolute value(p)2] calculated for the von Karman spectrum of temperature fluctuations agrees well with experimental data obtained in a laboratory experiment. PMID- 11386545 TI - Statistical moments of the sound field propagating in a random, refractive medium near an impedance boundary. AB - Propagation of a monochromatic sound field in a refractive and turbulent medium near an impedance boundary is considered. Starting from the parabolic equation for a moving medium and using the Markov approximation, a closed equation for the statistical moments of arbitrary order of the sound-pressure field is derived. Numerical methods for directly solving the first- and second-moment versions of this equation are formulated. The first-moment formulation is very similar to parabolic equations (PEs) that are now widely used to calculate sound fields for particular realizations of a random medium. The second-moment formulation involves a large, fringed tridiagonal matrix, which is solved using iterative refinement and Cholesky factorization. The solution is computationally intensive and currently restricted to low frequencies. As an example, the first and second moments are directly calculated for upwind and downwind propagation of a sound wave through a turbulent atmosphere. For these cases, predictions from the second moment PE were statistically indistinguishable from the result of 40 random trials calculated with a standard Crank-Nicholson PE, although the second-moment PE yielded smoother results due to its ensemble-average nature. PMID- 11386546 TI - Spectral broadening of sound scattered by advecting atmospheric turbulence. AB - Scattering and spectral broadening of a monochromatic sound wave by atmospheric turbulence that is flowing with a uniform constant horizontal wind is considered. The acoustic source and a detector are at rest and at different positions in a ground-fixed frame. Analytic expressions are derived for the sound pressure scattered to the detector by a single eddy. Since distances and the scattering angle change with time as the eddy flows through the scattering volume, the detector signal has time-dependent amplitude and frequency, for which general formulas are derived. A computer code is developed that calculates the scattered signal and its Fourier transform from a single eddy, or from a steady-state collection of eddies of many different scale lengths that represents isotropic homogeneous turbulence flowing with the wind. The code utilizes a time-shift algorithm that reduces the calculation time substantially. Several numerical results from this code are presented, including simulations of a recent experiment. The predicted spectral shape, including peak width and jaggedness, are in good agreement with experiment. PMID- 11386547 TI - Long range source localization from single hydrophone spectrograms. AB - A source near the deep sound channel axis excites mode groups (or paths) that involve both deep sound channel and boundary interacting propagation. Dispersion from a broadband source as measured on a single hydrophone can be used to estimate source range. Furthermore, modal group speeds have a functional transition when passing through purely refractive to boundary reflecting phase speed regions which, under certain conditions, provides additional arrival structure to aid in source localization. This additional arrival structure is in the form of a focal region in a spectrogram. Indeed, different data sets from the Acoustic Thermometry of the Ocean Climate (ATOC) Program [ATOC Consortium, Science 281, 1327-1332 (1998)] show that localization can be accomplished using this focal region and/or the overall dispersion properties as originally suggested fifty years ago [M. Ewing and J. L. Worzel, Geo. Soc. Am., Memoir 27 (1948)]. PMID- 11386548 TI - Excitation of T-phases by seafloor scattering. AB - T-phases excited by suboceanic earthquakes are classified into two types: abyssal phases which are excited near the earthquake epicenter at seafloor depths far below the SOFAR velocity channel, and slope T-phases which are excited at continental, or ocean island slopes and ridges at distances up to several hundreds of kilometers from the epicenter. In this article, it is demonstrated that approximate time-frequency characteristics of both classes of T-phase can be synthesized under the assumption that T-phases are excited by scattering from a rough seafloor. Seafloor scattering at shallow depths preferentially excites low order acoustic modes that propagate efficiently within the ocean sound channel minimum. At greater depths, scattering excites higher order modes which interact weakly with the seafloor along much of the propagation path. Using known variations in near-source bathymetry, T-phase envelopes are synthesized at several frequencies for several events south of the Fox Islands that excited both types of T-phase. The synthesized T-phases reproduce the main time vs frequency features of each type of arrival; a higher frequency, nearly symmetric arrival excited near the epicenter and a longer duration, lower frequency arrival excited near the continental shelf, with a peak amplitude at about 5 Hz. PMID- 11386549 TI - Acoustic attenuation in three-component gas mixtures--theory. AB - Vibrational relaxation accounts for absorption and dispersion of acoustic waves in gases that can be significantly greater than the classical absorption mechanisms related to shear viscosity and heat conduction. This vibrational relaxation results from retarded energy exchange between translational and intramolecular vibrational degrees of freedom. Theoretical calculation of the vibrational relaxation time of gases based on the theory of Landau and Teller [Phys. Z. Sovjetunion 10, 34 (1936); 1, 88 (1932): 2, 46 (1932)] and Schwartz et al. [J. Chem. Phys. 20, 1591 (1952)] has been applied at room temperature to ternary mixtures of polyatomic gases containing nitrogen, water vapor, and methane. Due to vibrational-translational and vibrational-vibrational coupling between all three components in ternary mixtures, multiple relaxation processes produce effective relaxation frequencies affecting the attenuation of sound. The dependence of effective relaxation frequencies and the attenuation on mole fractions of the constituents was investigated. The acoustic attenuation in a mixture that is primarily nitrogen is strongly dependent on the concentrations of methane and water vapor that are present. However, the attenuation in a mixture that is primarily methane is only weakly dependent on the concentrations of nitrogen and water vapor. The theory developed in this paper is applicable to other multicomponent mixtures. PMID- 11386550 TI - Elastic constants determination of anisotropic materials from phase velocities of acoustic waves generated and detected by lasers. AB - This paper describes a method which allows the synthesis of acoustic plane waves by means of laser ultrasonic techniques. When a laser line source radiates at the surface of an anisotropic material, intricate effects are observed on signals obtained by transmission experiments. The material characterization from such a set of signals is not a trivial task since it requires correctly selecting the data necessary for the elastic constants determination, by distinguishing them from all the parasitic phenomena. In the present paper, it is shown that a judicious sum of signals obtained for various positions of the line source at the surface of a sample leads to the synthesis of an acoustic plane wave for which the odd phenomena disappear. Moreover, by applying a constant delay between the various signals, acoustic plane waves can be synthesized with varied refracted angles. One of the advantages of this technique is to offer convenient access to the phase velocity, providing an easier determination of the elastic constants. This method is successively applied to recover the stiffness coefficients of a silicon crystal and of a composite material. The good agreement between the so recovered elastic constants and values given by other processes shows the contribution of such a method in the field of material characterization. PMID- 11386551 TI - The design, fabrication, and measured acoustic performance of a 1-3 piezoelectric composite Navy calibration standard transducer. AB - The design, fabrication, and acoustic calibration of a new 1-3 piezoelectric composite-based U.S. Navy standard (USRD-F82) are presented. The F82 dual array/parametric mode projector may be used as a reciprocal linear transducer, or may be used to exploit the nonlinear properties of the water to produce highly directional acoustic beams (4 to 3 deg) at relatively low frequencies (5 to 50 kHz, respectively). As a result of its wide bandwidth, a broad range of primary as well as secondary frequencies of operation is possible. In the linear mode of operation the transducer provides two separate arrays to be addressed topside for either transmit or receive applications. The two circular apertures are centered on the acoustic axis and have active diameters of 22.8 cm (9 in.) and 5.1 cm (2 in.). The smaller array aperture could be used to obtain broader acoustic beams at relatively high frequencies. Due to the absence of air-filled pressure release components, the transducer will operate over most ocean pressures and temperatures. A general description of the 1-3 piezoelectric composite-based transducer configuration and measured performance is presented. PMID- 11386552 TI - Maximum entropy approach for modeling random uncertainties in transient elastodynamics. AB - A new approach is presented for analyzing random uncertainties in dynamical systems. This approach consists of modeling random uncertainties by a nonparametric model allowing transient responses of mechanical systems submitted to impulsive loads to be predicted in the context of linear structural dynamics. The information used does not require the description of the local parameters of the mechanical model. The probability model is deduced from the use of the entropy optimization principle, whose available information is constituted of the algebraic properties related to the generalized mass, damping, and stiffness matrices which have to be positive-definite symmetric matrices, and the knowledge of these matrices for the mean reduced matrix model. An explicit construction and representation of the probability model have been obtained and are very well suited to algebraic calculus and to Monte Carlo numerical simulation in order to compute the transient responses of structures submitted to impulsive loads. The fundamental properties related to the convergence of the stochastic solution with respect to the dimension of the random reduced matrix model are analyzed. Finally, an example is presented. PMID- 11386553 TI - The matched-lag filter: detecting broadband multipath signals with auto- and cross-correlation functions. AB - Signal detection is considered for uncertain noise variance and a broadband source of unknown waveform and emission time. The signal travels to the receivers along paths with unknown delays. Using a new "matched-lag filter," the presence or absence of the signal is estimated from the auto- and cross-correlation functions of the receptions. Like a matched filter, correlation functions provide the first stage of gain in signal-to-noise ratio because the paths are assumed to be partially coherent. The second stage achieves additional gain by searching only over physically possible arrangements of signals in the auto- and cross correlation functions while excluding forbidden arrangements. These stages enable the matched-lag filter to behave like a matched filter within a matched filter. In an ideal case, simulations of the matched-lag filter yield probabilities of detection that are, with one and two receivers, 4.1 and 366 times, respectively, that obtained from the conventional energy detector at a false-alarm probability of 0.001. The matched-lag filter has applications to wireless communications and the detection of acoustic signals from animals, vehicles, ships, and nuclear blasts. The matched-lag filter more completely describes signal structure than stochastic detection and communication theories whose specified auto-correlation function does not prohibit forbidden arrangements. PMID- 11386554 TI - A compressive gammachirp auditory filter for both physiological and psychophysical data. AB - A gammachirp auditory filter was developed by Irino and Patterson [J. Acoust. Soc. Am. 101, 412-419 (1997)] to provide a level-dependent version of the linear, gammatone auditory filter, with which to explain the level-dependent changes in cochlear filtering observed in psychophysical masking experiments. In this 'analytical' gammachirp filter, the chirp varied with level and there was no explicit representation of the change in filter gain or compression with level. Subsequently, Carney et al. [J. Acoust. Soc. Am. 105, 2384-2391 (1999)] reviewed Carney and Yin's [J. Neurophysiol. 60, 1653-1677 (1988)] reverse-correlation (revcor) data and showed that the frequency glide of the chirp does not vary with level in their data. In this article, the architecture of the analytical gammachirp is reviewed with respect to cochlear physiology and a new form of gammachirp filter is described in which the magnitude response, the gain, and the compression vary with level but the chirp does not. This new 'compressive' gammachirp filter is used to fit the level-dependent revcor data reported by Carney et al. (1999) and the level-dependent masking data reported by Rosen and Baker [Hear. Res. 73, 231-243 (1994)]. PMID- 11386555 TI - Frequency glides in click responses of the basilar membrane and auditory nerve: their scaling behavior and origin in traveling-wave dispersion. AB - Frequency modulations (or glides), reported in impulse responses of both the auditory nerve and the basilar membrane, represent a change over time in the instantaneous frequency of oscillation of the response waveform. Although the near invariance of glides with stimulus intensity indicates that they are not the consequence of nonlinear or active processes in the inner ear, their origin has remained otherwise obscure. This paper combines theory with experimental data to explore the basic phenomenology of glides. When expressed in natural dimensionless form, glides are shown to have a universal form nearly independent of cochlear location for characteristic frequencies (CFs) above approximately 1.5 kHz (the "scaling region"). In the apex of the cochlea, by contrast, glides appear to depend strongly on CF. In the scaling region, instantaneous-frequency trajectories are shown to be approximately equal to the "inverse group delays" of basilar-membrane transfer functions measured at the same locations. The inverse group delay, obtained by functionally inverting the transfer-function group-delay versus-frequency curve, specifies the frequency component of a broadband stimulus expected to be driving the cochlear partition at the measurement point as a function of time. The approximate empirical equality of the two functions indicates that glides are closely related to cochlear traveling-wave dispersion and suggests that they originate primarily through the time dependence of the effective driving pressure force at the measurement location. Calculations in a one-dimensional cochlear model based on solution to the inverse problem in squirrel monkey [Zweig, J. Acoust. Soc. Am. 89, 1229-1254 (1991)] support this conclusion. In contrast to previous models for glides, which locate their origin in the differential build-up and decay of multiple micromechanical resonances local to each radial cross section of the organ of Corti, the model presented here identifies glides as the global consequence of the dispersive character of wave propagation in the cochlea. PMID- 11386556 TI - The effect of electrode configuration and duration of deafness on threshold and selectivity of responses to intracochlear electrical stimulation. AB - This report examines the effects of intracochlear electrode configuration and mode of stimulation (bipolar or monopolar) on neural threshold and spatial selectivity in the inferior colliculus (IC) of the cat. Single and multiunit IC recordings were made in three groups of animals; acutely deafened adults (controls), neonatally deafened animals studied at 6 to 18 months of age and neonatally deafened cats studied at 2.5 to 6.5 years. Response thresholds were plotted versus IC depth to measure the spatial distribution of responses. The response selectivity for each stimulating configuration was defined as the width of the resulting spatial tuning curve (STC) measured at 6 dB above threshold. Spiral ganglion cell (SG) survival was examined histologically in all neonatally deafened animals and correlated with physiological results. Animals studied at less than 1.5 years had SG densities of 23.5%-64.4% of normal (mean=42.7%) while animals studied at greater than 2.5 years had densities of 5.1%-18.3% of normal (mean=9.9%). Electrophysiological results include the following. (1) Monopolar thresholds were 7-8 dB lower than bipolar thresholds in the same animals. (2) Varying the configuration of bipolar contacts (measured as radial, offset radial and longitudinal pairs) did not systematically affect IC threshold in either controls or short-term neonatally deafened animals. In contrast, the long-term neonatally deafened animals showed a difference in threshold with each configuration. (3) The spatial distributions (Q(6 dB)) of responses to bipolar stimulation were approximately 40% more restricted than those for monopolar stimulation. (4) The spatial selectivity of neonatally deafened animals studied at ages up to 1.5 years was equal to that of control animals with normal auditory experience. However, selectivity was degraded in the older animals. (5) Selectivity was decreased in some animals with the longitudinal bipolar configuration and multiple response peaks were seen in several cases using this stimulus configuration. PMID- 11386557 TI - Emphasis of short-duration acoustic speech cues for cochlear implant users. AB - A new speech-coding strategy for cochlear implant users, called the transient emphasis spectral maxima (TESM), was developed to aid perception of short duration transient cues in speech. Speech-perception scores using the TESM strategy were compared to scores using the spectral maxima sound processor (SMSP) strategy in a group of eight adult users of the Nucleus 22 cochlear implant system. Significant improvements in mean speech-perception scores for the group were obtained on CNC open-set monosyllabic word tests in quiet (SMSP: 53.6% TESM: 61.3%, p<0.001), and on MUSL open-set sentence tests in multitalker noise (SMSP: 64.9% TESM: 70.6%, p<0.001). Significant increases were also shown for consonant scores in the word test (SMSP: 75.1% TESM: 80.6%, p<0.001) and for vowel scores in the word test (SMSP: 83.1% TESM: 85.7%, p<0.05). Analysis of consonant perception results from the CNC word tests showed that perception of nasal, stop, and fricative consonant discrimination was most improved. Information transmission analysis indicated that place of articulation was most improved, although improvements were also evident for manner of articulation. The increases in discrimination were shown to be related to improved coding of short-duration acoustic cues, particularly those of low intensity. PMID- 11386558 TI - The relationship between frequency selectivity and overshoot. AB - Under some conditions, threshold for a brief tone is higher at the onset of a broadband masker than it is if it is delayed from the onset of the masker. Evidence suggests that this "overshoot" is related to active processing in the auditory system. The present experiments examined this question, by measuring frequency selectivity under the same conditions in which overshoot was measured. The first experiment demonstrated that the growth of masking with masker level was approximately linear for a 1-kHz signal with or without a precursor (which was identical to the masker), and a 4-kHz signal with a precursor. For the 4-kHz signal with no precursor, an elevation in signal-to-masker ratio was seen at mid masker levels, relative to the other conditions. Frequency selectivity was then measured for a fixed-level signal, with and without a precursor. Relative frequency selectivity was highest for the 4-kHz signal with no precursor, lower for the 1-kHz signal with no precursor, and lowest for the 1- or 4-kHz signal with a precursor. The overshoot results and the frequency selectivity results would be consistent with stronger active processing at 4 kHz than at 1 kHz, and a decrease in active processing following a broadband noise precursor. PMID- 11386559 TI - The lower limit of melodic pitch. AB - An objective melody task was used to determine the lower limit of melodic pitch (LLMP) for harmonic complex tones. The LLMP was defined operationally as the repetition rate below which listeners could no longer recognize that one of the notes in a four-note, chromatic melody had changed by a semitone. In the first experiment, the stimuli were broadband tones with all their components in cosine phase, and the LLMP was found to be around 30 Hz. In the second experiment, the tones were filtered into bands about 1 kHz in width to determine the influence of frequency region on the LLMP. The results showed that whenever there was energy present below 800 Hz, the LLMP was still around 30 Hz. When the energy was limited to higher-frequency regions, however, the LLMP increased progressively, up to 270 Hz when the energy was restricted to the region above 3.2 kHz. In the third experiment, the phase relationship between spectral components was altered to determine whether the shape of the waveform affects the LLMP. When the envelope peak factor was reduced using the Schroeder phase relationship, the LLMP was not affected. When a secondary peak was introduced into the envelope of the stimuli by alternating the phase of successive components between two fixed values, there was a substantial reduction in the LLMP, for stimuli containing low frequency energy. A computational auditory model that extracts pitch information with autocorrelation can reproduce all of the observed effects, provided the contribution of longer time intervals is progressively reduced by a linear weighting function that limits the mechanism to time intervals of less than about 33 ms. PMID- 11386560 TI - A measure for predicting audibility discrimination thresholds for spectral envelope distortions in vowel sounds. AB - Both in speech synthesis and in sound coding it is often beneficial to have a measure that predicts whether, and to what extent, two sounds are different. This paper addresses the problem of estimating the perceptual effects of small modifications to the spectral envelope of a harmonic sound. A recently proposed auditory model is investigated that transforms the physical spectrum into a pattern of specific loudness as a function of critical band rate. A distance measure based on the concept of partial loudness is presented, which treats detectability in terms of a partial loudness threshold. This approach is adapted to the problem of estimating discrimination thresholds related to modifications of the spectral envelope of synthetic vowels. Data obtained from subjective listening tests using a representative set of stimuli in a 3IFC adaptive procedure show that the model makes reasonably good predictions of the discrimination threshold. Systematic deviations from the predicted thresholds may be related to individual differences in auditory filter selectivity. The partial loudness measure is compared with previously proposed distance measures such as the Euclidean distance between excitation patterns and between specific loudness applied to the same experimental data. An objective test measure shows that the partial loudness measure and the Euclidean distance of the excitation patterns are equally appropriate as distance measures for predicting audibility thresholds. The Euclidean distance between specific loudness is worse in performance compared with the other two. PMID- 11386561 TI - Revisiting relations between loudness and intensity discrimination. AB - A comparison is made between the variation of delta Ljnd with L (loudness), based on the beat-detection data of Riesz at 1 kHz [Phys. Rev. 31, 867-875 (1928)], and analogous relations obtained from a cross section of studies. Data analysis shows that only beat detection exhibits the degree of level-dependent variation in slope relating log (delta Ljnd) to log (L) described in a recent paper by Allen and Neely [J. Acoust. Soc. Am. 102, 3628-3646 (1997)]. Moreover, the slope variation determined from beat detection is not dependent on the detailed shape of the loudness function. The results imply that Allen and Neely's strong conclusions about the dependence of delta Ljnd on L are too tightly coupled to Riesz's methodology to be generally applicable. PMID- 11386562 TI - Infants' sensitivity to broadband noise. AB - Infants have higher pure-tone thresholds than adults. One explanation is that infants do not adopt the frequency-selective listening strategy that adults use when detecting tones. In contrast to other models of infants' immature sensitivity, the listening strategy account predicts that infants will be more sensitive to broadband sounds, relative to adults. Infants 7-9 months old were tested in two experiments to examine their sensitivity to broadband noise. Unmasked and masked thresholds for a 1000-Hz tone and for broadband noise were estimated adaptively for infants and adults using an observer-based behavioral procedure. The difference between infants and adults in unmasked threshold were 14 and 7 dB for tones and noise, respectively. The difference between infants and adults in masked threshold were 10 and 5 dB for tones and noise, respectively. Psychometric functions for detection of broadband noise were also obtained from some infants and adults. Infants' psychometric functions were similar to those obtained in tone detection with shallower slopes and lower upper asymptotes than adults'. This suggests that the relative improvement in infants' threshold for broadband noise is not due to greater attentiveness to the noise. A model of infants' sound detection invoking inattentiveness, listening strategy, and an unspecified source of internal noise may account for the characteristics of the infant psychometric function. PMID- 11386563 TI - Spatial release from informational masking in speech recognition. AB - Three experiments were conducted to determine the extent to which perceived separation of speech and interference improves speech recognition in the free field. Target speech stimuli were 320 grammatically correct but nonmeaningful sentences spoken by a female talker. In the first experiment the interference was a recording of either one or two female talkers reciting a continuous stream of similar nonmeaningful sentences. The target talker was always presented from a loudspeaker directly in front (0 degrees). The interference was either presented from the front loudspeaker (the F-F condition) or from both a right loudspeaker (60 degrees) and the front loudspeaker, with the right leading the front by 4 ms (the F-RF condition). Due to the precedence effect, the interference in the F-RF condition was perceived to be well to the right, while the target talker was heard from the front. For both the single-talker and two-talker interference, there was a sizable improvement in speech recognition in the F-RF condition compared with the F-F condition. However, a second experiment showed that there was no F-RF advantage when the interference was noise modulated by the single- or multi-channel envelope of the two-talker masker. Results of the third experiment indicated that the advantage of perceived separation is not limited to conditions where the interfering speech is understandable. PMID- 11386564 TI - Sound localization in the presence of one or two distracters. AB - Localizing a target sound can be a challenge when one or more distracter sounds are present at the same time. This study measured the effect of distracter position on target localization for one distracter (17 positions) and two distracters (21 combinations of 17 positions). Listeners were instructed to point to the apparent position of a train of 30-ms noise bursts, presented at 1 of 85 positions in virtual free field. A harmonic complex and a frequency-swept complex tone served as distracters. The two distracters were turned on 40 and 80 ms after the target onset, had temporal envelopes similar to that of the target, and did not overlap temporally with the target. Virtual sounds were synthesized with individual HRTFs. Localization performance degraded as the number of distracters increased from 0 to 2. When the horizontal distance between target and a single distracter was small (i.e., the interaural differences were almost the same), the influence on the apparent position was greater than when they were far apart. In the vertical dimension, there was not a systematic effect of distracter position on target localizability. However, there was a substantial increase in localization error for targets at high elevations (above 30 degrees) when distracters were present. PMID- 11386565 TI - Perceptual "vowel spaces" of cochlear implant users: implications for the study of auditory adaptation to spectral shift. AB - Cochlear implant (CI) users differ in their ability to perceive and recognize speech sounds. Two possible reasons for such individual differences may lie in their ability to discriminate formant frequencies or to adapt to the spectrally shifted information presented by cochlear implants, a basalward shift related to the implant's depth of insertion in the cochlea. In the present study, we examined these two alternatives using a method-of-adjustment (MOA) procedure with 330 synthetic vowel stimuli varying in F1 and F2 that were arranged in a two dimensional grid. Subjects were asked to label the synthetic stimuli that matched ten monophthongal vowels in visually presented words. Subjects then provided goodness ratings for the stimuli they had chosen. The subjects' responses to all ten vowels were used to construct individual perceptual "vowel spaces." If CI users fail to adapt completely to the basalward spectral shift, then the formant frequencies of their vowel categories should be shifted lower in both F1 and F2. However, with one exception, no systematic shifts were observed in the vowel spaces of CI users. Instead, the vowel spaces differed from one another in the relative size of their vowel categories. The results suggest that differences in formant frequency discrimination may account for the individual differences in vowel perception observed in cochlear implant users. PMID- 11386566 TI - Effects of delayed auditory feedback (DAF) on the pitch-shift reflex. AB - Changes in voice pitch auditory feedback to vocalizing subjects elicit compensatory changes in voice fundamental frequency (F0). The neural mechanisms responsible for this behavior involve the auditory and vocal-motor systems, collectively known as the audio-vocal system. Previous work [Burnett et al., J. Acoust. Soc. Am. 103, 3153-3161 (1998); Hain et al., Exp. Brain Res. 130, 133-141 (2000); Larson et al., J. Acoust. Soc. Am. 107, 559-564 (2000)] indicated that this system operates using negative feedback to cancel out low-level errors in voice F0 output. By introducing delays in the auditory feedback pathway, we hoped to transiently "open" the feedback loop and learn which components of the response are most closely related to the timing of the auditory feedback signal. Subjects were presented with pitch-shift stimuli that were paired with a delay of 0, 50, 100, 200, 300, or 500 ms. Delayed auditory feedback did not affect voice F0 response latency or magnitude, but it delayed the timing of later parts of the response. As a further test of the audio-vocal control system, a second experiment was conducted in which delays of 100, 200, or 300 ms were combined with stimuli having onset velocities of 1000 or 330 cents/s. Results confirmed earlier reports that the system is sensitive to velocity of stimulus onset. A simple feedback model reproduced most features of both experiments. These results strongly support previous suggestions that the audio-vocal system monitors auditory feedback and, through closed-loop negative feedback incorporating a delay, adjusts voice F0 so as to cancel unintentional small magnitude fluctuations in F0. PMID- 11386567 TI - Dynamic consequences of differences in male and female vocal tract dimensions. AB - Phonetic differences between male and female speakers are generally considered in terms of the static acoustic and perceptual consequences of different articulatory dimensions. This article investigates the dynamic acoustic and articulatory implications of differences in mean male and female vocal tract dimensions. The temporal acoustic consequences of time-varying twin-tube resonators of different dimensions are explored, and the possible implications for human speech production are considered. Empirical support for the theoretical predictions is sought by investigating the kinematic and acoustic patterns in diphthong productions from 26 female and 22 male speakers in the University of Wisconsin X-ray Microbeam Speech Production Database. Aside from expected acoustic differences, the shape of male and female formant tracks plotted in Bark space is found to be very similar. Male and female patterns of tongue movement, however, are found to be very dissimilar. The mean male diphthong, defined by the tracks of four midsagittal pellets, is characterized by greater pellet excursions, higher pellet speed, and consistently larger dorso-palatal strictures than its female counterpart. The empirical findings suggest that gender-specific dynamic behavior could be an important factor in accounting for nonuniform vowel system differences, but at the same time having more wide-ranging implications for transitional phenomena and undershoot. PMID- 11386568 TI - Linear degrees of freedom in speech production: analysis of cineradio- and labio film data and articulatory-acoustic modeling. AB - The following contribution addresses several issues concerning speech degrees of freedom in French oral vowels, stop, and fricative consonants based on an analysis of tongue and lip shapes extracted from cineradio- and labio-films. The midsagittal tongue shapes have been submitted to a linear decomposition where some of the loading factors were selected such as jaw and larynx position while four other components were derived from principal component analysis (PCA). For the lips, in addition to the more traditional protrusion and opening components, a supplementary component was extracted to explain the upward movement of both the upper and lower lips in [v] production. A linear articulatory model was developed; the six tongue degrees of freedom were used as the articulatory control parameters of the midsagittal tongue contours and explained 96% of the tongue data variance. These control parameters were also used to specify the frontal lip width dimension derived from the labio-film front views. Finally, this model was complemented by a conversion model going from the midsagittal to the area function, based on a fitting of the midsagittal distances and the formant frequencies for both vowels and consonants. PMID- 11386570 TI - A cross-language comparison of /d/-/th/ perception: evidence for a new developmental pattern. AB - Previous studies have shown that infants perceptually differentiate certain non native contrasts at 6-8 months but not at 10-12 months of age, whereas differentiation is evident at both ages in infants for whom the test contrasts are native. These findings reveal a language-specific bias to be emerging during the first year of life. A developmental decline is not observed for all non native contrasts, but it has been consistently reported for every contrast in which language effects are observed in adults, In the present study differentiation of English /d-th/ by English- and French-speaking adults and English- and French-learning infants at two ages (6-8 and 10-12 months) was compared using the conditioned headturn procedure. Two findings emerged. First, perceptual differentiation was unaffected by language experience in the first year of life, despite robust evidence of language effects in adulthood. Second, language experience had a facilitative effect on performance after 12 months, whereas performance remained unchanged in the absence of specific language experience. These data are clearly inconsistent with previous studies as well as predictions based on a conceptual framework proposed by Burnham [Appl. Psycholing. 7, 201-240 (1986)]. Factors contributing to these developmental patterns include the acoustic properties of /d-th/, the phonotactic uniqueness of English /th/, and the influence of lexical knowledge on phonetic processing. PMID- 11386569 TI - The relationship between spectral characteristics and perceived hypernasality in children. AB - The purpose of this study was to quantify perceived hypernasality in children. One-third octave spectra of the isolated vowel [i] were obtained from 32 children with cleft palate and 5 children without cleft palate. Four experienced listeners rated the severity of hypernasality of the 37 speech samples using a 6-point equal-appearing interval scale. When the average 1/3-octave spectra from the hypernasal group and the normal resonance group were compared, spectral characteristics of hypernasality were identified as increased amplitudes between F1 and F2 and decreased amplitudes in the region of F2. Based on the findings of the children's speech, 36 speech samples with manipulated spectral characteristics were used to minimize the influences of voice source characteristics on perceived hypernasality. Multiple regression analysis revealed a high correlation (R = 0.84) between the amplitudes of 1/3-octave bands (1 k, 1.6 k, and 2.5 kHz) and the perceptual ratings. Increased amplitudes of bands between F1 and F2 (1 k, 1.6 kHz) and decreased amplitude of the band of F2 (2.5 kHz) was associated with an increasing perceived hypernasality. These results suggest that the amplitudes of the three 1/3-octave bands are appropriate acoustic parameters to quantify hypernasality in the isolated vowel [i]. PMID- 11386571 TI - The effect of intensity perturbations on speech intelligibility for normal hearing and hearing-impaired listeners. AB - Hearing-impaired listeners are known to suffer from reduced speech intelligibility in noise, even if sounds are above their hearing thresholds. This study examined the possible contribution of reduced acuity of intensity coding to this problem. The "distortion-sensitivity model" was used: the effect of reduced acuity of auditory intensity coding on intelligibility was mimicked by an artificial distortion of the speech intensity coding, and the sensitivity to this distortion for hearing-impaired listeners was compared with that for normal hearing listeners. Stimuli (speech plus noise) were wavelet coded using a Gaussian wavelet (1/4-octave bandwidth). The intensity coding was distorted by multiplying the modulus of each wavelet coefficient by a random factor. Speech reception thresholds (SRTs) were measured for various degrees of intensity perturbation. Hearing-impaired listeners were classified as suffering from suprathreshold deficits if intelligibility of undistorted speech was worse than predicted from audibility by the speech intelligibility index model [ANSI, ANSI S3.5-1997 (1997)]. Hearing-impaired listeners without suprathreshold deficits were as sensitive to the intensity distortion as the normal-hearing listeners. Hearing-impaired listeners with suprathreshold deficits appeared to be less sensitive. Results indicate that reduced acuity of auditory intensity coding may be a factor underlying reduced speech intelligibility in noise for the hearing impaired. PMID- 11386573 TI - Acoustic-phonetic features for the automatic classification of fricatives. AB - In this article, the acoustic-phonetic characteristics of the American English fricative consonants are investigated from the automatic classification standpoint. The features studied in the literature are evaluated and new features are proposed. To test the value of the extracted features, a statistically guided, knowledge-based, acoustic-phonetic system for the automatic classification of fricatives in speaker-independent continuous speech is proposed. The system uses an auditory-based front-end processing system and incorporates new algorithms for the extraction and manipulation of the acoustic phonetic features that proved to be rich in their information content. Classification experiments are performed using hard-decision algorithms on fricatives extracted from the TIMIT database continuous speech of 60 speakers (not used in the design/training process) from seven different dialects of American English. An accuracy of 93% is obtained for voicing detection, 91% for place of articulation detection, and 87% for the overall classification of fricatives. PMID- 11386572 TI - Age-related changes in detecting a mistuned harmonic. AB - The effects of age on discriminating simultaneous sounds were investigated by comparing the hearing threshold in detecting a mistuned harmonic in young, middle aged, and older adults. The stimuli were complex sounds containing multiple harmonics, one of which could be "mistuned" so that it was no longer an integer multiple of the fundamental. Older adults had higher thresholds than middle-aged or young adults. The effect of age was greater for short than for long duration sounds and remained even after controlling for hearing sensitivity. The results are consistent with an age-related decline in parsing simultaneous auditory events, which may contribute to the speech perception difficulties in the elderly. PMID- 11386574 TI - Experimental validation of the use of Kramers-Kronig relations to eliminate the phase sheet ambiguity in broadband phase spectroscopy. AB - The technique of broadband phase spectroscopy proposed in 1978 by Sachse and Pao [J. Appl. Phys. 49, 4320-4327 (1978)] determines the phase velocity as a function of frequency from the Fourier transforms of a received reference and through sample signal. Although quite successful, this approach can be influenced by an ambiguity in the phase velocity calculation which stems from the boundedness of the inverse tangent operation used to calculate phase. Several empirical approaches to resolve the phase ambiguity have been reported. An alternative approach that has not previously been considered appeals to the causal nature of the measurements. This article experimentally validates a method which uses the causally consistent Kramers-Kronig relations to eliminate the ambiguity in phase spectroscopy-derived phase velocity calculations. Broadband pulse and narrow-band tone burst measurements were performed on three gelatin-based phantoms containing different concentrations of graphite particles (0%, 10%, and 20% by volume). The phantoms were constructed to have attenuation coefficients which vary approximately linear-with-frequency, a dependence exhibited by many soft tissues. The narrow-band phase velocity measurements do not suffer from a phase ambiguity, and thus they serve as a "gold standard" against which the broadband phase velocity measurements are compared. The experimental results illustrate that using the Kramers-Kronig dispersion relations in conjunction with phase spectroscopy-derived phase velocity measurements is an effective means by which to resolve the phase sheet ambiguity in broadband phase spectroscopy. PMID- 11386575 TI - Simulations of the thermo-acoustic lens effect during focused ultrasound surgery. AB - Laboratory measurements of soft tissue properties show a dependence of background propagation properties on temperature. For typical focused ultrasound surgery (FUS) applications, only the slow variations in tissue background parameters need to be accounted for when computing the outcome of a FUS sonication. The cumulative effect of slowly varying sound speed has been referred to in the literature as a thermal lens, or a thermo-acoustic lens because of its beam distorting properties. An algorithm to solve the coupled acoustic-thermal problem is described, and numerical results are presented to illustrate the effects of dynamic sound-speed profiles in layered tissues undergoing FUS. The results of simulations in liver with and without a fat layer indicate that the thermal acoustic interaction results in more complex dynamics in FUS than a simple model will predict. Both the size and the position of the lesions predicted from the simulations are affected by the thermo-acoustic lens effect. However, the overall effect from short sonications at high power from sharply focused single element sources (F-no. from 0.8 to 1.3) around 1 MHz similar to those used in clinical setups is found to be small. PMID- 11386576 TI - Vocal behavior of male sperm whales: why do they click? AB - Off Kaikoura, New Zealand, we recorded individually identified male sperm whales (Physeter macrocephalus) for entire dive cycles in order to investigate vocal behavior of individual whales and to examine possible functions of sperm whale clicks. In our study, sperm whales were almost always silent at the surface. They consistently started clicking within 25 s after fluking-up and diving. During the first 10 s of clicking, interclick intervals were significantly correlated with water depths at the location of fluke-up. The first "creak" was produced on average 7.5 min into a dive. Interclick intervals usually decreased substantially before clicks turned into "creaks." The highest click rate recorded in this study was 90.9 click/s, and clicks-within-creaks were much shorter than "usual clicks" (mean of 3.6 ms versus 17 to 30 ms). The number of creaks per minute of dive and the length of a dive were significantly correlated. On average, sperm whales were silent for the last 3.6 min before surfacing. Short sequences of "surface clicks" (3 to 8 metallic clicks with mean interclick interval of 5.5 s) were often produced at the end of a dive (in 57% of the dives), but their function remains puzzling. The results of this study suggest that usual clicks and creaks are both used for echolocation purposes, the former to gather information about acoustically reflective features and the latter to detect prey. PMID- 11386578 TI - Improved description of shock wave evolution in media with frequency power law dependent attenuation. PMID- 11386581 TI - The effect of speechreading on masked detection thresholds for filtered speech. PMID- 11386582 TI - Evaluation of speech intelligibility with the coordinate response measure. PMID- 11386583 TI - Heart failure in older people: the epidemic we had to have. PMID- 11386584 TI - Preventing transmission of HIV from mothers to babies in Australia. PMID- 11386585 TI - Compliance with clinical guidelines for blood transfusion practice: how can changes be maintained? PMID- 11386586 TI - The costs of urinary incontinence. PMID- 11386587 TI - Chronic heart failure in Australian general practice. The Cardiac Awareness Survey and Evaluation (CASE) Study. AB - OBJECTIVES: To investigate the frequency and general practitioner awareness of patients with chronic heart failure (CHF), and to evaluate a cardiac algorithm and document cardiac investigations performed in establishing this diagnosis. DESIGN AND SETTING: Between March and August 1998, consecutive patients aged 60 years and older presenting to their GP were assessed. In patients previously diagnosed with CHF, aetiology and diagnostic assessments were documented. In patients with suspected CHF (by a standardised algorithm, based on World Health Organization guidelines), further investigations and GP diagnosis were recorded. PATIENTS: 80 consecutive patients were assessed by each of 341 GPs throughout Australia, reflecting the Australian metropolitan/rural population mix of 1996. This provided a total of 22060 evaluable patients. MAIN OUTCOME MEASURES: Estimated numbers of patients with CHF in general practice (previously and newly diagnosed); major aetiological factors; use of ancillary diagnostic tests; drugs prescribed. RESULTS: CHF was diagnosed in 2905 of 22060 patients (13.2%) (2485 previously diagnosed and 420 newly diagnosed). Major aetiological factors were ischaemic heart disease and hypertension. Echocardiography had been performed in 64% of previously diagnosed patients, but was performed in only 22% of possible CHF patients. Angiotensin-converting enzyme (ACE) inhibitors were prescribed in 58.1% of patients with CHF. Patients with evidence of left ventricular dysfunction were more likely to have received ACE inhibitors. CONCLUSIONS: CHF appears to be very common in the elderly, based on GP diagnosis of the condition. Of 100 patients aged 60 years and over presenting to their GP, two new cases of CHF will be detected using a simple clinical algorithm in conjunction with appropriate diagnostic tests. ACE inhibitors appear to be underutilised. PMID- 11386588 TI - Durability of response to a targeted intervention to modify clinician transfusion practices in a major teaching hospital. AB - OBJECTIVES: To determine the durability of a successful intervention to modify clinician transfusion practices, and to compare current transfusion practices in the "intervention" hospital with those in a hospital with no intervention. DESIGN: Prospective, descriptive study. SETTING: Two major metropolitan teaching hospitals--Royal Melbourne Hospital and Western Hospital, Footscray. SUBJECTS: Consecutive patient transfusion episodes for red cells, platelets and fresh frozen plasma (FFP). OUTCOME MEASURES: Appropriateness of transfusion according to intervention guidelines; comparison of inappropriate transfusion rates before the intervention, immediately after the intervention and 3 years after the intervention. Comparison of inappropriate transfusion rates in intervention and non-intervention hospitals. RESULTS: Inappropriate transfusion rates 3 years after the intervention were 20% for red cells, 27% for platelets, and 43% for FFP. These were significantly higher than equivalent rates reported immediately after the intervention. Inappropriate transfusion rates at the non-intervention hospital were comparable (26% for red cells, 36% for platelets and 52% for FFP). CONCLUSION: Appropriate clinician transfusion practices have proven difficult to sustain 3 years after hospital guideline generation and promotion. A "gate keeping" role by hospital blood bank staff proved impractical in the long term. PMID- 11386589 TI - Use of interventions for reducing mother-to-child transmission of HIV in Australia. AB - OBJECTIVE: To describe the extent and outcome of use of interventions for reducing the risk of HIV transmission from mother to child in Australia. DESIGN: National surveillance for perinatal exposure to HIV. PARTICIPANTS AND SETTING: Notified cases of HIV infection in women in Australia and their perinatally exposed children, 1982-1999. OUTCOME MEASURES: Trends over time in use of interventions (antiretroviral therapy in pregnancy, elective caesarean delivery and avoidance of breastfeeding) and perinatally acquired HIV infection. RESULTS: By 31 March 2000, 204 children were reported as having been born in 1982-1999 to 162 women whose HIV infection had been diagnosed by 31 December 1999. The child's HIV infection status was established for 182 (89.2%); the mother's HIV infection was diagnosed antenatally in 91 of these cases (50%). Among women diagnosed antenatally, use of elective caesarean delivery and antiretroviral therapy in pregnancy increased significantly, from 3% and 14% by women whose children were born in 1982-1993, to 21% (P=0.01) and 88% (P<0.001), respectively, by women whose children were born in 1994-1999. Most women (95%) diagnosed antenatally avoided breastfeeding their children. The percentage of infected children born to women diagnosed antenatally declined from 26% among children born in 1982-1993 to 19% among those born in 1994-1999. The percentage of infected children was significantly lower among those whose mothers used antiretroviral therapy in pregnancy (11% versus 36%; P=0.03). CONCLUSION: Antiretroviral use in pregnancy, elective caesarean delivery and avoidance of breastfeeding have been effective interventions for reducing the risk of mother-to-child HIV transmission in Australia. While the rate of perinatal HIV transmission has declined, it remains high in comparison with rates reported from other industrialised countries. PMID- 11386590 TI - Poisoning with the recreational drug paramethoxyamphetamine ("death"). AB - OBJECTIVE: To describe the clinical features of paramethoxyamphetamine (PMA; "death") poisoning and to compare these with those of people with self-reported "ecstasy" poisoning. DESIGN: Retrospective casenote review. PARTICIPANTS AND SETTING: 22 patients who presented to the Emergency Department of the Royal Adelaide Hospital (RAH), a major metropolitan teaching hospital, between 1 January 1996 and 31 December 1998 with PMA poisoning identified through urine drug screens; and 61 patients with self-reported ecstasy poisoning between 1 September 1997 and 31 December 1998 found through the hospital databases. RESULTS: Patients with PMA poisoning presented with tachycardia (64%), hyperthermia (temperature > 37.5 degrees C; 36%), coma (41%), seizures (32%), arrhythmias (23%), and QRS intervals > or = 100 ms (50%) with greater frequency and often greater severity than those with self-reported ecstasy poisoning. Two patients with PMA poisoning presented with severe hypoglycaemia (blood glucose level, < 1.5 mmol/L) accompanied by hyperkalaemia (K+ concentration, > 7.5 mmol/L). CONCLUSIONS: At our hospital, PMA poisonings accounted for most of the severe reactions among people who believed they had taken ecstasy. Hypoglycaemia and hyperkalaemia may be specific to PMA poisoning. PMA toxicity should be suspected with severe or atypical reactions to "ecstasy", and confirmed by chromatographic urine drug screens. PMID- 11386591 TI - Economic costs of urinary incontinence in community-dwelling Australian women. AB - OBJECTIVE: To estimate the economic cost of urinary incontinence in community dwelling Australian women aged 18 years and over for the year 1998. DESIGN: Extrapolation of data from studies of women with incontinence to the Australian population of women aged 18 years and over in 1998. MAIN OUTCOME MEASURES: Estimated prevalence of urinary incontinence in 1998, and estimated cost in Australian dollars of resource use and personal costs related to management of incontinence. RESULTS: An estimated 1835628 community-dwelling women over the age of 18 years had urinary incontinence in 1998. The total annual cost of this urinary incontinence is estimated at $710.44 million, or $387 per incontinent woman, comprising $338.47 million in treatment costs and $371.97 million in personal costs. An estimated 60% of women with incontinence in 1998 were aged 40 years or over. Assuming the prevalence of incontinence remains constant and, allowing for inflation, we project that the total annual cost in 20 years' time will be $1267.85 million, 93% ($1.18 billion) of which will constitute costs associated with women aged over 40 years. CONCLUSIONS: Urinary incontinence imposes a considerable drain on Australian healthcare resources. More research is needed to understand the magnitude of the problem and potential gains from continence promotion. PMID- 11386593 TI - Maximising the uptake of evidence into clinical practice: an information economics approach. AB - Evidence such as systematic reviews or clinical practice guidelines are information products, and clinicians are consumers of those products; their current proliferation but low uptake by consumers indicates an information oversupply. The costs and benefits of accessing and applying information are at least as important as are the costs and benefits of the treatments the information describes. In the same way that a citation index is a measure of the impact of a scientific paper, an evidence uptake index could measure effectiveness of evidence products in a clinical population. The uptake of evidence-based medicine may be hampered by the perceived high cost of changing to it. At present, most costs are borne by individual clinicians, but individual benefits for clinicians are downplayed in favour of population benefits. Specific strategies to increase evidence uptake into practice include decreasing the "cost of ownership"; increasing the direct or perceived value of evidence resources in routine practice; and customising evidence to suit different users, tasks and clinical contexts. PMID- 11386592 TI - Guidelines for management of patients with chronic heart failure in Australia. AB - Chronic heart failure (CHF) affects approximately 1% of people aged 50-59 years, and this high prevalence increases dramatically with age. CHF is a common reason for hospital admission and general practitioner consultation in the elderly. Common causes of CHF are ischaemic heart disease, hypertension and idiopathic dilated cardiomyopathy. Diagnosis of CHF is based on clinical features and objective measurement of ventricular function (eg, echocardiography). Management is directed at prevention, retarding disease progression, relief of symptoms and prolonging survival. Non-pharmacological approaches include exercise, home-based support and risk-factor modification. Angiotensin-converting enzyme (ACE) inhibitors are the cornerstone of pharmacological therapy to prevent disease progression and prolong survival. beta-Blockers prolong survival when added to ACE inhibitors in symptomatic patients. Diuretics provide symptom relief and restoration or maintenance of euvolaemia. Spironolactone, angiotensin II receptor antagonists and digoxin may be useful in some patients. Surgical approaches in highly selected patients may include myocardial revascularisation, insertion of devices and cardiac transplantation. PMID- 11386594 TI - Healthcare rationing, patient rights and the law. PMID- 11386595 TI - Legionella pneumophila: not just pneumonia. PMID- 11386596 TI - Stains on the caring mantle. PMID- 11386597 TI - Bupropion and serum sickness-like reaction. PMID- 11386599 TI - Non-O1 Vibrio cholerae: a fatal cause of sepsis in northern Australia. PMID- 11386598 TI - Foot-propelled scooter injuries during the Sydney 2000 Olympic Games period. PMID- 11386600 TI - The use of non-steroidal anti-inflammatory drugs in subacute thyroiditis. PMID- 11386601 TI - Latex allergy and food handlers. PMID- 11386602 TI - Non-cardiac chest pain. PMID- 11386603 TI - Asthma clinics in general practice: what is the evidence? PMID- 11386604 TI - Optimization of preanalytical conditions and analysis of plasma glucose. 1. Impact of the new WHO and ADA recommendations on diagnosis of diabetes mellitus. AB - The new diagnostic criteria for type 2 diabetes from the American Diabetes Association (ADA) and World Health Organization (WHO) recommend measurements on plasma and a lowering of the glucose threshold for diabetes by 0.8 mmol/L. This narrows the distance between the upper end of the reference limit and the discriminatory level to a degree where analytical quality becomes critical. The quality demands for the preanalytical and analytical phase and their consequences on diagnostic performance have to be established in the new technical system, measuring in plasma rather than in capillary whole blood. Because of the instability of glucose in blood samples it is necessary to clarify the influence of different preanalytical and analytical factors on the number of false-positive and false-negative classifications. Thus the aim of the present study was to find optimal conditions for sampling, additives, storage, transport and analysis of plasma glucose combining feasibility with an analytical bias close to zero and a within-imprecision around 1%. We have documented the analytical performance of the method itself and its traceability to an international standard. The preanalytical conditions, such as influence of antiglycolytic agent NaF, conditions for plasma separation, storage temperature and storage time before and after plasma separation were investigated. In conclusion, we recommend that blood should be drawn in tubes containing heparin and NaF and kept on ice water for not more than 1 h until centrifugation at minimum 1000 x g for 10 min. The plasma is then stable for at least 48 h at room temperature. PMID- 11386605 TI - Plasma glucose reference interval in a low-risk population. 2. Impact of the new WHO and ADA recommendations on the diagnosis of diabetes mellitus. AB - The aim of the study was to establish a reference interval of fasting venous plasma glucose (FPG) from healthy individuals. A prospective modified cross sectional population-based study was made with random selection of 2100 persons in age-stratified groups > or = 18 years identified from the local Personal Identification Register. The invitation was accepted by 755 persons, of which 726 aged 18-92 years were eligible. They did not have a diabetes diagnosis, were non pregnant and capable of fasting for 8 h. All participants filled in a questionnaire on medical risk factors. Blood for the FPG and haemoglobin Alc (HbAlc) measurements was drawn in accordance with a standardized procedure. A total of 302 participants carried diabetes risk indicators and were ruled out. The FPG concentrations in the remaining low-risk population (n=424) was ln Gaussian distributed. The FPG 97.50 centile in this group was 6.4 mmol/L (95% CI: 6.3-6.5 mmol/L), in contrast to the WHO and ADA theoretical limit of 6.1 mmol/L. Their diagnostic decision limit of 7.0 mmol/L FPG corresponded to the 99.86 centile of the FPG reference distribution (95% CI: 6.8-7.1). Subclassification of the reference population showed increasing FPG with increasing BMI and age and was higher in men than in women. The study determined the FPG 95% interfractile reference interval in a healthy population. The interval in glucose concentration between the 97.5 centile of the reference interval and the ADA-WHO diagnostic limit is very narrow. The clinical application of the diagnostic discriminator and the interpretation of the WHO-ADA grey zone from 6.1 to 7.0 mmol/L FPG may consequently be biased because of poorly defined limits and influence from BMI, age and gender. PMID- 11386606 TI - Evaluation of systematic and random factors in measurements of fasting plasma glucose as the basis for analytical quality specifications in the diagnosis of diabetes. 3. Impact of the new WHO and ADA recommendations on diagnosis of diabetes mellitus. AB - On behalf of the Danish Society of Clinical Endocrinology and the Danish Society of Clinical Chemistry we were commissioned to evaluate the influence of analytical and pre-analytical systematic and random factors on the diagnosis of diabetes, in order to provide a tool for conclusions on the analytical quality specifications needed to diagnose diabetes. A systems analysis was performed in accordance with the principles for evaluation of analytical quality specifications. The clinical setting was defined--diagnosis of diabetes in accordance with the WHO and ADA criteria with determination of fasting plasma glucose concentration (FPG) > or =7.0 mmol/L in two independent samples--with well-documented data on In (loge)-Gaussian distribution of reference values from a low-risk population and values for within-subject biological variation taken from the literature. An investigation was made of the consequences for the clinical setting of assumed errors related to the measurement of FPG. Four approaches were investigated for a single sampling and measurement and also for two independent samples: one showing the percentage of healthy individuals who had values > or = 7.0 mmol/L, one illustrating the origin of biological set points for results > or = 7.0 mmol/L, one showing the risk of being measured > or =7.0 mmol/L when the biological set-point is known, and one showing the combined bias and imprecision for assumed percentages of false-positive (FP), defined as measurements > or = 7.0 mmol/L for the low-risk population and false-negative (FN), defined as measurements <6.4 mmol/L (the upper reference limit) for diabetics. This leaves a "grey zone" which includes the upper part of low-risk individuals, and defined by ADA and WHO as "impaired fasting glucose" (IFG). In the analysis, increasing systematic and random errors (combined analytical and pre-analytical) were assumed, and for each error condition the fractions of FP and FN were calculated. This gave plots from which the combined allowable systematic and random errors could be read off for pre-determined clinically acceptable fractions of FP and FN. The analysis does not distinguish between pre analytical and analytical errors, as specified information on one of these is needed for specification of the other. The investigation provides a reliable basis for estimation of the needed analytical quality, and thereby for decisions about analytical quality specifications for analysis of FPG in relation to diagnosis of diabetes under optimized pre-analytical and analytical conditions. Consequences of deviations from these ideal conditions are illustrated in the figures, and should be considered for the different approaches with different performance conditions. PMID- 11386607 TI - Alpha-fetoprotein in plasma and serum of healthy adults: preanalytical, analytical and biological sources of variation and construction of age-dependent reference intervals. AB - Alpha-fetoprotein (AFP) is a tumor marker for hepatomas and germ cell tumors, and the serum concentration has prognostic significance in other diseases. We examined the normal serum concentration of AFP in adults and sources of variation in the immunochemical variation of AFP. The serum concentration of the tumor marker alpha-fetoprotein (S-AFP) was log-normally distributed in 284 adult blood donors. S-AFP increased with age (p < 10(-7)), whereas no gender-related difference was found. Reference intervals (95-interpercentile) were constructed for persons < or =40 years (0.60-9.30 kIU/L) and >40 years (1.40 12.60 kIU/L). The concentration of AFP was significantly, albeit slightly, higher in serum than in plasma, whereas hemolysis, pretreatment with KCl and food intake did not influence S-AFP. S-AFP only changed 6% when measured twice 2 months apart (p=0.04). Three enzyme immunoassays, using three different anti-AFP monoclonal antibodies for detection, were compared and two assays gave S-AFP values significantly higher, 2.8% (p=0.03) and 19.0% (p<10(-4)), than the other assay. Thus, the choice of antibody may influence the result of immunochemical concentration determination. This can be explained by the existence of conformational variants of AFP with different antibody reactivities, and calls for careful standardization of monoclonal antibodies used in assays for AFP. With broad population reference ranges and slight intra-personal variation, the most effective reference range for S-AFP is previous values obtained in the same person. PMID- 11386608 TI - Renal elimination of protein S-100beta in pigs with acute encephalopathy. AB - BACKGROUND: Protein S-100beta is an established biochemical marker for cerebral injury in serum. For the further interpretation and possible use of S-100beta serum measurements in acute hepatic encephalopathy, renal elimination of S 100beta was measured in pigs with elevated S-100beta levels due to hepatic encephalopathy. METHODS: Eighteen female Norwegian Landrace pigs were randomly allocated to either hepatic devascularization (n=13) or sham operation (n=5). Repeated samples from the common carotid artery, right renal vein, and urine were simultaneously drawn for S-100beta analysis, using the Sangtec100 Liamat immunoassay. RESULTS: In hepatic devascularized pigs, arterial serum levels of S 100beta increased from 0.96+/-0.04 microg/L (mean +/- SEM) at t = 0h to 1.74+/ 0.11 microg/L (mean +/- SEM) at t = 5 h. Urinary excretion increased simultaneously from 8.48+/-3.66 ng/h (mean +/- SEM) to 20.4+/-9.54 ng/h (mean +/- SEM), while renal arterial-venous fluxes for both kidneys increased from 1022+/ 404 ng/h (mean +/- SEM) to 2444+/-590 ng/h (mean +/- SEM). CONCLUSIONS: Increased arterial S-100beta levels in pigs with acute hepatic encephalopathy are not a result of decreased renal elimination. The large difference between the renal arterial venous S-100beta concentrations and the urinary excretion of S-100beta indicate that renal metabolism is the major route of elimination. PMID- 11386609 TI - Comparison of diagnostic performance of cardiac troponin I on the IMMULITE system with other automated troponin I assays in minor myocardial damage. AB - An analytical and clinical evaluation of cardiac troponin I (cTnI) on the IMMULITE system is presented. The assay results were compared with those of the Stratus II and the Dimension RxL-HM. A between-run imprecision CV < 20% was found at a cTnI concentration of 0.23 microg/L (functional limit of detection). On the basis of a reference study including 215 patients without ischemic heart disease (97.5th percentile: 0.294 microg/L) and 36 patients clinically classified as having stable angina pectoris (<0.22 microg/L) a preliminary cutoff level of 0.3 microg/L was defined. Assay linearity, sample stability, influence of sample material and method comparison studies were performed. In patients with Duchenne's disease, chronic hemodialysis treatment, pulmonary embolism, coronary artery bypass surgery and minimally cardiac surgery the cTnI results of the IMMULITE agreed better with the Dimension RxL-HM than with the Stratus II data. Of 142 samples from patients with unstable angina 67 samples were classified as cTnI positive with the IMMULITE, 76 with the Dimension RxL-HM, and 62 with the Stratus II. In conclusion, the new assay is sensitive for the determination of cTnI and easy to perform within 45 min. PMID- 11386610 TI - Urinary excretion of diazepam metabolites in healthy volunteers and drug users. AB - Urinary excretion profiles of diazepam metabolites were investigated. The subjects were healthy volunteers receiving one single 10-mg dose of diazepam or drug abusers starting a prison sentence. Urinary excretion of metabolites was analysed by immunological screening, liquid chromatography and gas chromatography mass spectrometry. Relating the metabolite concentration to creatinine concentration in the specimens decreased sample-to-sample variations. In some cases such correction could protect a subject from erroneous accusations of a new intake. PMID- 11386611 TI - The mucosa-associated lymphoid tissue structure, function, and derangements. AB - Nearly 50% of the immune cells in the body lie just beneath the moist mucosal surfaces at intestinal and extra-intestinal sites. The study of this mucosal immune system in response to shock and to route and type of nutrition provides a cogent explanation for the reduced incidence of pneumonia with enteral feeding. Changes in immune cell mass and function are associated with deterioration of previously established immunity at mucosal surfaces, especially the respiratory tract. By understanding the mechanisms involved in this breakdown, therapeutic strategies can be developed to reduce septic complications in critical illness. PMID- 11386612 TI - Differential release of interleukines 6, 8, and 10 in cerebrospinal fluid and plasma after traumatic brain injury. AB - Traumatic brain injury (TBI) is characterized by a high mortality which is largely determined by the initial cerebral trauma, secondary brain injury or indirectly during a Multiple Organ Dysfunction Syndrome (MODS). Therefore, we analyzed IL-6, IL-8, and IL-10 in cerebrospinal fluid (CSF) and in plasma with respect to blood-brain barrier (BBB) integrity in 29 patients suffering from isolated TBI. IL-6 and IL-8 were significantly increased compared to baseline levels early after trauma in CSF and plasma. In all patients CSF IL-6 and IL-8 were found to be higher than corresponding plasma levels. IL-10 in plasma was significantly increased above control plasma values, however, without a significant difference to the corresponding CSF values. BBB dysfunction was temporary present in 23 patients. Significant correlations between BBB dysfunction and cytokines were not found. Thus, alterations of the BBB seems not to influence the distribution pattern of interleukines in CSF and plasma after trauma. PMID- 11386614 TI - Injury induces alterations in T-cell NFkappaB and AP-1 activation. AB - The immune dysfunction that occurs after severe injury involves major changes in T-cell-mediated immunity resulting in suppressed T-helper 1 (Th1) type responses and increased or persistent T-helper 2 (Th2) type cytokine production. Since little is known about what signaling pathways are responsible for this injury induced phenotypic shift in T-cells, we undertook this study to address the molecular basis for injury effects on T-helper cell subset cytokine expression. Experiments were designed to test whether diminished IL-2 gene expression after thermal injury coincided with changes in the induction of IL-2 gene regulatory transcription factors. Electrophoretic mobility shift assays (EMSA) were used to screen for nuclear expression of changes of the IL-2 gene transcription factors. Our findings revealed that changes in mitogen-stimulated T-cell AP-1 and NFkappaB factor activation correlated directly with defective mitogen-induced IL-2 mRNA expression. We determined that there was a loss of nuclear AP-1 activation and changes in NFkappaB factor activation at 9 days after injury. T-cell nuclear extracts prepared from sham injured mice showed induction of NFkappaB2 (p52) and RelA (p65) containing NFkappaB EMSA complexes, while we detected no RelA or NFkappaB2 in EMSA complexes using T-cell nuclear extracts prepared from burn injured mice. Instead, these NFkappaB EMSA complexes contained mostly NFkappaB1 (p50). Western immunoblot analysis confirmed defective nuclear RelA translocation. Taken together, these results indicate that T-cell NFkappaB and AP 1 activation pathways may be involved in the injury-induced changes in T-cell cytokine production and the immune deviation that occurs after injury. PMID- 11386613 TI - Effect of dopexamine on hepatic metabolic activity in patients with septic shock. AB - Hepato-splanchnic metabolic activity is seen to be related to regional blood flow and oxygen/substrate availability in patients with sepsis. Catecholamines, which may modulate metabolic activity perse, are common to stabilize hemodynamics. We studied the effect of a dopexamine-induced increase in splanchnic blood flow (Qspl) on regional metabolic rate in 10 patients with septic shock requiring norepinephrine to maintain mean arterial pressure (>60 mmHg). Splanchnic blood flow was determined using the indocyanine-green method with hepatic venous sampling. We determined the hepato-splanchnic lactate, pyruvate, alanine, and glutamine turnover and the lactate/pyruvate and ketone body ratio as well as the endogenous glucose production (EGP) using the stable isotope approach. Qspl increased from 0.86 (0.79-1.15) to 0.96 (0.92-1.33) L/min/m2, not influencing any parameter of metabolic activity. We speculate that this finding is due to altered beta-adrenoreceptor-mediated thermogenic effects due to the interplay of different beta-sympathomimetics at the receptor site. PMID- 11386615 TI - Kupffer cells and neutrophils as paracrine regulators of the heme oxygenase-1 gene in hepatocytes after hemorrhagic shock. AB - Heme oxygenase (HO) plays a pivotal role for the maintenance of liver blood flow and hepatocellular integrity after hemorrhagic shock. We investigated the role of Kupffer cells and neutrophils as paracrine modulators of hepatocellular HO-1 gene expression in a rat model of hemorrhage and resuscitation. Male Sprague-Dawley rats (n = 6-10/group) were anesthetized (pentobarbital, 50 mg/kg intraperitonal) and subjected to hemorrhagic shock (mean arterial blood pressure: 35 mmHg for 60 min) or a sham protocol. Based on the time course of HO-1 gene expression, the effect of various antioxidants, Kupffer cell blockade [gadolinium chloride (GdCl3); 10 mg/kg; 24 h prior to hemorrhage or dichloromethylene diphosphonate (Cl2MDP); 1 mg/kg; 2 days prior to hemorrhage], or neutrophil depletion (vinblastine, 0.5 mg/kg, 5 days prior to hemorrhage) on induction of the HO-1 gene was assessed at 5 h of resuscitation, i.e., the time point of maximal induction. Kupffer cell blockade and antioxidants abolished HO-1 mRNA and protein induction after hemorrhage, while neutrophil depletion failed to affect hepatocellular HO-1 gene expression. In addition, Kupffer cell blockade aggravated hepatocellular injury. N-formyl-methionine-leucyl-phenylalanin (fMLP) induced a substantial influx of neutrophils into the liver but failed to induce hepatocellular HO-1 mRNA expression. These data suggest that Kupffer cells but not neutrophils induce an adaptive hepatocellular stress response after hemorrhage and resuscitation. Oxygen-free radicals released by Kupffer cells may serve as paracrine regulators of a hepatocellular stress gene which is necessary to maintain liver blood flow and integrity under stress conditions. PMID- 11386616 TI - Aminoethyl-isothiourea inhibits the increase in plasma endothelin-1 caused by serogroup A streptococci and prolongs survival in rat peritoneal sepsis. AB - To elucidate the possible roles of nitric oxide (NO), endothelin-1 (ET-1), and reactive oxygen species (ROS) in the pathophysiology of serogroup A streptococcal (GAS) peritoneal sepsis, we investigated the effects of aminoethylisothiourea (AE ITU), an inducible NO synthase (iNOS) inhibitor, and a ROS scavenger, and the ET 1 receptor antagonist bosentan. In rats, live GAS inocula, 3 x 10(8) and 1 x 10(9) cfu/kg, entailed a 24-h mortality of 10% and 90%, respectively. GAS caused increases in tissue iNOS activity (9 h), in serum nitrite/nitrate (9-24 h), and in intracellular leukocyte ROS levels (3-6 h). These changes were all prevented by the pre-treatment with AE-ITU. A novel finding was that AE-ITU also prevented the GAS-induced marked increase in plasma ET-1 at 6 h. Short-term (7-h) survival was improved by both AE-ITU and by bosentan. The mechanism(s) for the beneficial effects of AE-ITU may possibly be a combined mode of action; iNOS inhibition, ROS scavenging, and inhibition of the increase in plasma ET-1 caused by GAS. PMID- 11386617 TI - Aminoethyl-isothiourea in gram-positive shock: an inhibitor of inducible nitric oxide synthase or a jack-of-all-trades? PMID- 11386618 TI - Aminoethyl-isothiourea inhibits leukocyte production of reactive oxygen species and proinflammatory cytokines induced by streptococcal cell wall components in human whole blood. AB - The incidence of severe invasive disease caused by serogroup A streptococci (GAS) is increasing, and to elucidate the role of streptococcal cell wall components in the inflammatory response, human whole blood was stimulated with lipoteichoic acid (LTA, 0.005-50 microg/mL) and peptidoglycan (10 and 100 microg/ml) from Streptococcus pyogenes. Both stimulants increased dose dependently the leukocyte release of cytokines many thousand fold: tumor necrosis factor alpha (0 to 158,000+/-4,900 pg/mL), interleukin (IL)-1beta (85+/-56 to 31,000+/-4,600 pg/mL), IL-6 (30+/-11 to 34,800+/-15,000 pg/mL), and IL-8 (300+/-150 to 29,000+/-14,000 pg/mL). Intracellular leukocyte levels of reactive oxygen species (ROS) as measured by flow cytometry increased 15-20 fold, from 25 to 400-500 mean fluorescence intensity. Aminoethyl-isothiourea (AE-ITU), a relatively selective inhibitor of the inducible nitric oxide synthase (iNOS) and a ROS scavenger, reduced the cytokine production by 70-100%, and intracellular leukocyte ROS levels by 50-70% (all P < 0.05). The non-selective NOS inhibitor N-nitro-L arginine methyl ester (L-NAME) did not affect intracellular ROS levels, but it caused a moderate selective inhibition of IL-8 production. Leukocyte NO production (measured up to 36 h) was not enhanced by LTA, peptidoglycan, inactivated streptococci, or cytokine combinations. The mechanisms for the anti inflammatory effects of AE-ITU may be through a reduction of intracellular ROS levels, or through a direct effect on signal transduction, whereas NO modulation is an unlikely mechanism. PMID- 11386619 TI - Proinflammatory effects of bacterial lipoprotein on human neutrophil activation status, function and cytotoxic potential in vitro. AB - Bacterial lipoprotein (BLP) is the most abundant protein in gram-negative bacterial cell walls, heavily outweighing lipopolysaccharide (LPS). Herein we present findings demonstrating the potent in vitro effects of BLP on neutrophil (PMN) activation status, function, and capacity to transmigrate an endothelial monolayer. PMNs are the principal effectors of the initial host response to injury or infection and constitute a significant threat to invading bacterial pathogens. The systemic inflammatory response syndrome (SIRS) is characterised by significant host tissue injury mediated, in part, by uncontrolled regulation of PMN cytotoxic activity. We found that BLP-activated human PMN as evidenced by increased CD11b/CD18 (Mac-1) expression. Up-regulation of PMN Mac-1 in response to BLP occurred independently of membrane-bound CD14 (mCD14). A similar up regulation of intercellular adhesion molecule-1 (ICAM-1) on endothelial cells was observed whilst E-Selectin expression was unaffected. PMN transmigration across a human umbilical vein endothelial cell (HUVEC) monolayer was markedly increased after treating either PMN's or HUVEC independently with BLP. This increased transmigration did not occur as a result of any direct effect of BLP on HUVEC monolayer permeability, assessed objectively using the passage of FITC-labeled Dextran-70. BLP primed PMN for enhanced respiratory burst and superoxide anion production in response to PMA, but did not influence phagocytosis of opsonized Escherichia coli. BLP far exceeds LPS as a gram-negative bacterial wall component, these findings therefore implicate BLP as an additional putative mediator of SIRS arising from gram-negative infection. PMID- 11386620 TI - Morphologic changes of red blood cells during hemorrhagic shock replicate changes of aging. AB - Blood loss leads to the reduction in vitality of red blood cells (RBCs). However, the changes in morphology at different stages of hemorrhagic shock have not been studied. Thus, the aim of this study was to identify and quantitate the sequence of morphological changes in RBCs during hemorrhage. This study was performed on 15 adult inbred dogs. Blood samples were taken before hemorrhage, when the mean arterial pressure reached 40 mm Hg (initial stage of shock), and at a mean arterial pressure level of 20 mm Hg (decompensated stage of shock). The volume of blood removed averaged 33.6+/-8.9 and 55.1+/-6.9 mL/kg, respectively. Evaluation of RBC morphology was performed by computerized light microscopic morphometry and scanning electron microscopy. At the early stage of hemorrhage the number of "young-appearing" RBCs with large visible surface areas (41-50 microm2) increased from 17.7%+/-3.1% to 26.6%+/-3.5% (P < 0.05). Concomitantly, the number of "old appearing" RBCs with small visible surface area (20-30 microm2) significantly decreased from 5.3%+/-2.7% to 2.7%+/-2.3% (P < 0.01). At the stage of decompensated blood loss, the opposite phenomenon was observed. The number of "old-appearing" RBCs increased to 8.2%+/-1.1% (P < 0.01), whereas the number of "young-appearing" RBCs decreased to 12.3%+/-4.2% (P< 0.01). The changes in visible surface area of RBCs was accompanied by significant alterations in their shape. The percentage of abnormal shaped RBCs increased from 8.9%+/-1.1% before the hemorrhage to 36.4%+/-5.8% at the stage of decompensated hemorragic shock (P < 0.01). Thus, during the late decompensated stage of hemorrhagic shock, RBCs assume shape and surface area changes that are similar to those seen in aging. These changes in RBC size and shape may be due to the effects of shock-induced oxidative stress. PMID- 11386621 TI - Opiate modulation of hemodynamic, hormonal, and cytokine responses to hemorrhage. AB - The aim of the present study was to examine the role of opiate receptor activation in modulating the hemodynamic, neuroendocrine, and tissue (lung and spleen) cytokine responses to fixed pressure (40 mm Hg) hemorrhage. Chronically catheterized, conscious unrestrained non-heparinized male Sprague-Dawley rats were pretreated with either naltrexone (15 mg/kg intraperitoneally in 0.5 mL of saline) or saline (0.5 mL) 15 min prior to hemorrhage followed by fluid resuscitation with Ringer's lactate. Animals were sacrificed at completion of the 60-min resuscitation period. Blood loss required to achieve mean arterial blood pressure (MABP) of 40 mm Hg was higher in naltrexone-treated animals than in time matched saline controls (4.4+/-0.2 versus 3.7+/-0.2 mL/100 g BW, P< 0.05). Hemorrhage increased plasma levels of corticosterone (30%) and ACTH (3-fold) within 15 min. Naltrexone prevented the hemorrhage-induced rise in corticosterone without affecting the rise in ACTH. Hemorrhage increased beta-endorphin levels (4 fold) and produced an immediate (5 min) and progressive increase in circulating epinephrine and norepinephrine levels reaching values that were 50- and 20-fold, respectively, higher than basal. Pre-treatment with naltrexone did not alter the time course or magnitude of the hemorrhage-induced increases in plasma beta endorphin or catecholamines. Hemorrhage increased lung and spleen content of TNF (60%), IL-1alpha (300%), IL-6 (40%-60%), and IL-10 (80%) above values of time matched sham control animals. Pre-treatment with naltrexone blunted the magnitude of the increases in tissue cytokine content in response to a given blood loss. These results indicate that endogenous opiates modulate the hemodynamic instability, neuroendocrine, and cytokine responses to hemorrhagic shock. PMID- 11386622 TI - Combination therapy that targets secondary pulmonary changes after abdominal trauma. AB - After abdominal trauma, the lung is susceptible to secondary injury caused by acute neutrophil (PMN) sequestration and alveolar capillary membrane disruption. Adenosine is an endogenous anti-inflammatory metabolite that decreases PMN activation. AICAR ([5-amino-1-[beta-D-ribofuranosyl]imidazole-4 carboxamide]riboside) is the prototype of a novel class of anti-inflammatory drugs that increase endogenous adenosine. After trauma, AICAR administration has been shown to decrease secondary lung injury in models of hemorrhagic shock with delayed lipopolysaccharide challenge and pulmonary contusion. However, early suppression of PMN activation could worsen outcomes after penetrating abdominal trauma. We hypothesized that, after penetrating abdominal trauma, the ideal resuscitation strategy would involve early, short-lived suppression of PMN activation to minimize secondary lung injury, followed by later enhancement of PMN chemotaxis and phagocytosis [using granulocyte colony-stimulating factor (G CSF)] to lessen late septic complications. G-CSF has not been shown to potentiate PMN mediated pulmonary reperfusion injury. Swine were subjected to cecal ligation/incision and hemorrhagic shock (trauma), followed by resuscitation with shed blood, crystalloid, and either G-CSF, a combination of G-CSF and AICAR, or 0.9% normal saline. At 72 h, bronchoalveolar lavage (BAL) leukocyte counts and protein concentration were determined, and lung tissue analysed for myeloperoxidase (MPO, a measure of PMN infiltration) and microscopic pathology. Analysis of BALs revealed a significant increase protein concentrations and in white blood cell and PMN infiltration (P< 0.05) following trauma. These acute changes were not exacerbated by G-CSF, but were reversed by combined AICAR + G CSF, which implicates a physiologic role for adenosine. This suggests that combination therapy may have beneficial effects on the lung after trauma. PMID- 11386623 TI - Risk-adjusted monitoring of binary surgical outcomes. AB - A graphical procedure suitable for prospectively monitoring surgical performance is proposed. The approach is based on accumulating evidence from the outcomes of all previous surgical patients in a series using a new type of cumulative sum chart. Cumulative sum procedures are designed to "signal" if sufficient evidence has accumulated that the surgical failure rate has changed substantially. In this way, the chart rapidly detects deterioration (or improvement) in surgical performance while not overreacting to the expected fluctuations due to chance. Through the use of a likelihood-based scoring method, the cumulative sum procedure is adapted so that it adjusts for the surgical risk of each patient estimated preoperatively. The procedure is therefore applicable in situations where it is desirable to adjust for a mix of patients. Signals of the chart lead to investigations of the cause and to the timely introduction of remedial measures designed to avoid unnecessary future failures. PMID- 11386624 TI - Decision-case mix model for analyzing variation in cesarean rates. AB - This article contributes a decision-case mix model for analyzing variation in c section rates. Like recent contributions to the literature, the model systematically takes into account the effect of case mix. Going beyond past research, the model highlights differences in physician decision making in response to obstetric factors. Distinguishing the effects of physician decision making and case mix is important in understanding why c-section rates vary and in developing programs to effect change in physician behavior. The model was applied to a sample of deliveries at a hospital where physicians exhibited considerable variation in their c-section rates. Comparing groups with a low versus high rate, the authors' general conclusion is that the difference in physician decision tendencies (to perform a c-section), in response to specific obstetric factors, is at least as important as case mix in explaining variation in c-section rates. The exact effects of decision making versus case mix depend on how the model application defines the obstetric condition of interest and on the weighting of deliveries by their estimated "risk of Cesarean." The general conclusion is supported by an additional analysis that uses the model's elements to predict individual physicians' annual c-section rates. PMID- 11386625 TI - Preference for equity as a framing effect. AB - BACKGROUND: In previous studies, the authors found that most people, given a fixed budget, would rather offer a less effective screening test to 100% of a Medicaid population than a more effective test to 50% of the population. In a subsequent study, the authors found that the number of people preferring the less effective screening test was dramatically reduced when the percentage of Medicaid enrollees receiving it was less than 100. In this article, 2 new studies are reported that explore whether people's preferences for equity versus efficiency are susceptible to a framing effect. METHODS: In 2 studies, the authors presented subjects with multiple scenarios involving screening tests that vary in the proportion of people who could receive the tests within a budget constraint and the number of people whose lives each test would save. Across scenarios, the proportion of Medicaid enrollees who could receive each test was varied, as was the question of whether scenarios involved Medicaid enrollees from the same or a different state. In addition, the authors varied the order in which subjects received the scenarios. RESULTS: In the 1st study, people's preferences for equity over efficiency varied significantly depending on the way situations were framed. Preference for equity was stronger when the more widely distributed choice covered the entire population than when it covered only half the population (P < 0.001). In addition, people's preferences were susceptible to order effects, with preference for equity being significantly stronger when the 1st scenario received by subjects involved 1 screening test that could be offered to the entire population (P < 0.001). In the 2nd study, preferences for equity over efficiency diminished even when the different framings were descriptions of identical circumstances--preference for equity was significantly reduced when the population to be screened was framed broadly, in terms of the percentage of patients across 2 states who could receive testing rather than narrowly, in terms of the percentages of patients in 1 state who could receive testing (P = 0.04). CONCLUSION: Policy planners should be careful about accepting public preferences for equity over efficiency at face value, because such preferences can be dramatically influenced by framing effects and order effects. PMID- 11386626 TI - Do nonpatients underestimate the quality of life associated with chronic health conditions because of a focusing illusion? AB - BACKGROUND: A number of studies show that the general public often estimates that the quality of life (QOL) associated with various health conditions is worse than patients say it is. These studies raise the possibility that people overestimate the impact that unfamiliar health conditions will have on their quality of life. One possible reason people overestimate this is because they are susceptible to a focusing illusion--when asked to imagine themselves in unfamiliar circumstances, people overestimate the emotional impact of those features of their life that would change. METHODS: The authors surveyed members of the general public to test the hypothesis that their QOL ratings of hypothetical health conditions would be higher (indicating a better quality of life) after thinking about how the health condition would affect a broad range of life domains. Across 3 experiments, the authors varied the health conditions people were asked to consider (either paraplegia, below-the-knee amputation, or partial blindness), the life domains they were asked to consider, the response mode with which they evaluated how each health condition would affect each life domain, whether subjects rated the health condition before and after considering life domains or only after, and whether subjects rated their own current quality of life first. RESULTS: Across 3 experiments, using 10 different questionnaire versions, only 1 instance was found in which subjects' ratings were significantly higher after thinking about the effect of the health condition on life domains than before, and the magnitude of this increase was small. CONCLUSION: It could not be established that a focusing illusion contributes significantly to the discrepancy in QOL ratings of patients and nonpatients. Further research should explore other factors that could contribute to the discrepancy or other ways of testing for the influence of a focusing illusion. PMID- 11386627 TI - What should be reported in a methods section on utility assessment? AB - BACKGROUND: The measurement of utilities, or preferences, for health states may be affected by the technique used. Unfortunately, in papers reporting utilities, it is often difficult to infer how the utility measurement was carried out. PURPOSE: To present a list of components that, when described, provide sufficient detail of the utility assessment. METHODS: An initial list was prepared by one of the authors. A panel of 8 experts was formed to add additional components. The components were drawn from 6 clusters that focus on the design of the study, the administration procedure, the health state descriptions, the description of the utility assessment method, the description of the indifference procedure, and the use of visual aids or software programs. The list was updated and redistributed among a total of 14 experts, and the components were judged for their importance of being mentioned in a Methods section. RESULTS: More than 40 components were generated. Ten components were identified as necessary to include even in an article not focusing on utility measurement: how utility questions were administered, how health states were described, which utility assessment method(s) was used, the response and completion rates, specification of the duration of the health states, which software program (if any) was used, the description of the worst health state (lower anchor of the scale), whether a matching or choice indifference search procedure was used, when the assessment was conducted relative to treatment, and which (if any) visual aids were used. The interjudge reliability was satisfactory (Cronbach's alpha = 0.85). DISCUSSION: The list of components important for utility papers may be used in various ways, for instance, as a checklist while writing, reviewing, or reading a Methods section or while designing experiments. Guidelines are provided for a few components. PMID- 11386628 TI - Choice-matching preference reversals in health outcome assessments. AB - BACKGROUND: Health outcome utility assessments generally assume procedural invariance. Preference reversals violating procedural invariance occur in economic scenarios when the assessment process shifts from a choice to a fill-in the-blank task. PURPOSE: To determine if similar reversals occur in utility assessments. METHODS: One hundred thirty-six volunteer subjects completed 6 preference assessments of 4 personal health scenarios. Patients responded to otherwise identical tasks using either choice or fill-in-the-blank processes in a randomized crossover design. The authors determined the percentage of subjects preferring, or inferred to prefer, a given choice. RESULTS: Preference reversals occurred in all assessment scenarios. CONCLUSIONS: These preference reversals are a potential source of confusion for utility assessment and informed consent. They could be manipulated to achieve ends other than the best interest of patients. Anchoring or the prominence hypothesis may explain these findings. PMID- 11386629 TI - Bayesian assessment of sample size for clinical trials of cost-effectiveness. AB - The authors present an analysis of the choice of sample sizes for demonstrating cost-effectiveness of a new treatment or procedure, when data on both cost and efficacy will be collected in a clinical trial. The Bayesian approach to statistics is employed, as well as a novel Bayesian criterion that provides insight into the sample size problem and offers a very flexible formulation. PMID- 11386630 TI - Women's views on breast cancer risk and screening mammography: a qualitative interview study. AB - BACKGROUND: To promote informed decision making about mammography, clinicians are urged to present women with complete, relevant information about breast cancer and screening. Understanding women's current beliefs may help guide such efforts by uncovering misunderstandings, conceptual gaps, and areas of concern. OBJECTIVE: The authors sought to learn how women view breast cancer, their personal risk of breast cancer, and how screening mammography affects that risk. METHODS: Forty-one open-ended semistructured telephone interviews with women selected from a national database by quota sampling to ensure a wide range in demographics of the participants. RESULTS: Almost all respondents viewed breast cancer as a uniformly progressive disease that begins in a silent curable form (typically found by mammograms) and, unless treated early, invariably grows, spreads, and kills. Some women felt that any abnormality found must be treated, even if it was not malignant. None had heard of potentially nonprogressive cancers, and when informed, most felt that the uncertain prognosis of such lesions reinforced the need to find and treat disease as soon as possible. Women expressed a wide range of views about their personal risk of breast cancer. Although some saw breast cancer as a central threat to their health, many others cited heart disease, other cancers, violence, and trauma as greater concerns. Most recognized the importance of "uncontrollable" factors for breast cancer such as age, sex, family history, and genetics. However, other "controllable" factors with little or no demonstrated link to breast cancer (e.g., smoking, diet, toxic exposures, "bad attitudes") were given equal or greater prominence, suggesting that many women feel considerable personal responsibility for their level of breast cancer risk. Similarly, although women recognized that mammography was not perfect, almost all believed that failure to have mammograms put one at risk for premature and preventable death. When asked how mammography worked, almost all repeated the message that "early detection saves lives," suggesting that advanced cancer (and perhaps most cancer deaths) reflected a failure of early detection. The belief in the benefit of early detection was so strong that some women advocated scaring other women into getting mammograms because it is "better to be safe than sorry." CONCLUSIONS: Women view breast cancer as a uniformly progressive disease rarely curable unless caught early. The exaggerated importance many attribute to a variety of controllable factors in modifying personal risk and the "danger" seen in failing to have mammograms may lead women diagnosed with breast cancer to blame themselves. PMID- 11386632 TI - Industrial engineering and the practice of medicine. PMID- 11386631 TI - Patient-centered decision making: a view of the past and a look toward the future. PMID- 11386633 TI - Law and ethics. PMID- 11386634 TI - Amphetamine and N-acetylamphetamine incorporation into hair: an investigation of the potential role of drug basicity in hair color bias. AB - To elucidate the role of drug basicity in the preferential incorporation of certain drugs into dark hair rather than light hair, Long-Evans rats were dosed with amphetamine or its non-basic analogue N-acetylamphetamine (N-AcAp) and their hair evaluated for drug content. Rats were shaved prior to dosing. On the 14th day after dosing, hair from the same area that was shaved prior to dosing was shaved and collected. After the addition of amphetamine-d3 or N-AcAp-d3 as an internal standard, hair samples (20 mg) were digested in 1M NaOH at 37 degrees C. Digested solutions were then extracted with n-butyl chloride/chloroform (4:1, v/v). After drying and reconstituting, samples were injected onto a ThermoQuest TSQ liquid chromatography-tandem mass spectrometry instrument for analysis. Black hair from rats dosed with amphetamine (n = 8) was found to contain 6.44 +/- 1.31 (SD) ng amphetamine/mg hair. White hair from the same rats contained 2.04 +/- 0.58 ng amphetamine/mg hair. In contrast, no difference in N-AcAp content was found between black hair (0.87 +/- 0.08 ng N-AcAp/mg hair) and white hair (0.83 +/- 0.15 ng N-AcAp/mg hair) from rats dosed with N-AcAp (n = 8). PMID- 11386635 TI - Rapid determination of cyanide and azide in beverages by microdiffusion spectrophotometric method. AB - A rapid screening method was developed for the determination of the toxic volatile anions, cyanide and azide, in beverages. This method consisted of a microdiffusion extraction combined with spectrophotometry using the Konig cyanide reaction and ferric azide complex formation in conjugation with cerium azide oxido-reduction. The time required to achieve full recovery in the extraction of hydrogen cyanide and hydrazoic acid from samples was considerably shortened by increasing the diffusion temperature from 25 degrees C to 40 degrees C. The time required to achieve saturated color development in the Konig cyanide reaction was also shortened by increasing incubation temperature to 40 degrees C. The interference in both azide color reactions was examined for volatile compounds. Cyanide interfered only in the case of ferric azide complex formation. Sulfide, sulfate, nitrite, and acetic acid interfered in both the color reactions. The established method gave a detection limit of 6 microM for cyanide and 0.5mM for azide, and it required only 1 h to determine both anions. Cyanide and azide disappeared by evaporation from beverages during 25 degrees C storage under open conditions in a pH-dependent manner as a function of their respective pKa values of 9.2 and 4.6. PMID- 11386636 TI - Screening procedure for detection of non-steroidal anti-inflammatory drugs and their metabolites in urine as part of a systematic toxicological analysis procedure for acidic drugs and poisons by gas chromatography-mass spectrometry after extractive methylation. AB - Non-steroidal anti-inflammatory drugs (NSAIDs) are widely used as analgesic and anti-rheumatic drugs, and they are often misused. A gas chromatographic-mass spectrometric (GC-MS) screening procedure was developed for their detection in urine as part of a systematic toxicological analysis procedure for acidic drugs and poisons after extractive methylation. The compounds were separated by capillary GC and identified by computerized MS in the full-scan mode. Using mass chromatography with the ions m/z 119, 135, 139, 152, 165, 229, 244, 266, 272, and 326, the possible presence of NSAIDs and their metabolites could be indicated. The identity of positive signals in such mass chromatograms was confirmed by comparison of the peaks underlying full mass spectra with the reference spectra recorded during this study. This method allowed the detection of therapeutic concentrations of acemetacin, acetaminophen (paracetamol), acetylsalicylic acid, diclofenac, diflunisal, etodolac, fenbufen, fenoprofen, flufenamic acid, flurbiprofen, ibuprofen, indometacin, kebuzone, ketoprofen, lonazolac, meclofenamic acid, mefenamic acid, mofebutazone, naproxen, niflumic acid, phenylbutazone, suxibuzone, tiaprofenic acid, tolfenamic acid, and tolmetin in urine samples. The overall recoveries of the different NSAIDs ranged between 50 and 80% with coefficients of variation of less than 15% (n = 5), and the limits of detection of the different NSAIDs were between 10 and 50 ng/mL (S/N = 3) in the full-scan mode. Extractive methylation has proved to be a versatile method for STA of various acidic drugs, poisons, and their metabolites in urine. It has also successfully been used for plasma analysis. PMID- 11386637 TI - Severe intoxication with the veterinary tranquilizer xylazine in humans. AB - Xylazine (Rompun, Proxylaz) is a veterinary tranquilizing agent. A case of self injection of 1.5 g xylazine by a 27-year-old farmer is reported. He subsequently became comatose, hypotensive, bradycardic, and mildly glycemic. An intensive supportive therapy including intubation and ventilation was required. The patient made a full recovery over the next 30 h. The largest concentrations measured were 4.6 mg/L in plasma, 446 mg/L in gastric fluid, and 194 mg/L in urine. The calculated plasma half-life was 4.9 h. Kinetic data correlated with clinical symptoms. Qualitative and quantitative analyses of xylazine were done by thin layer chromatography, gas chromatography-mass spectrometry, and high-performance liquid chromatography. These methods allow the detection of small amounts substance in stomach, plasma, and urine. Liquid-liquid extraction was used for the isolation of drug. The sensitvity is high, and with these methods, a rapid analysis is possible. Xylazine intoxications in humans are rare. We describe the management of acute poisoning and present a review of xylazine toxicity in humans. PMID- 11386638 TI - HPLC with simultaneous UV and reductive electrochemical detection at the hanging mercury drop electrode: a highly sensitive and selective tool for the determination of benzodiazepines in forensic samples. AB - A versatile, sensitive, and selective high-performance liquid chromatographic (HPLC) procedure for the determination of common benzodiazepines and some of their most frequently occurring metabolites in forensic samples was developed and optimized with respect to effective and rapid sample preparation and high selectivity of the analytical assay. The optimized method includes liquid-liquid extraction of the drugs with chloroform followed by isocratic reversed-phase chromatography on a LiChrospher-100 RP-8ec column (150 x 4.6-mm i.d.) with a mobile phase consisting of 0.03 mol/L acetate buffer (pH 4.6)/acetonitrile (55:45, v/v). The use of dual-mode detection made up of UV-detection (250 nm) in series with reductive electrochemical detection (-1.4 V vs. Ag/AgCl) at the hanging mercury drop electrode permits the detection and quantitation of benzodiazepines even in degraded samples with higher selectivities than usually reached with conventional HPLC techniques. Depending on the actual benzodiazepine species, detection limits are in the range of 2.0 to 14.1 ng/mL. Mean recovery values of the drugs from blood range from 82 to 92%; within-day and day-to-day repeatabilities typically lie between 3 and 9%. Several case work examples demonstrate the high selectivity and remarkably low matrix sensitivity of the described assay. PMID- 11386639 TI - Profiles of urine samples taken from Ecstasy users at Rave parties: analysis by immunoassays, HPLC, and GC-MS. AB - The abuse of the designer amphetamines such as 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy) is increasing throughout the world. They have become popular drugs, especially at all-night techno dance parties (Raves), and their detection is becoming an important issue. Presently, there are no MDMA- or MDA-specific immunoassays on the market, and detection of the designer amphetamines is dependent upon the use of commercially available amphetamine assays. The success of this approach has been difficult to assess because of the general unavailability of significant numbers of samples from known drug users. The objectives of the present study are to characterize the drug content of urine samples from admitted Ecstasy users by chromatographic methods and to assess the ability of the available amphetamine/methamphetamine immunoassays to detect methylenedioxyamphetamines. We found that, when analyzed by high-performance liquid chromatography with diode-array detection (HPLC-DAD), 64% of 70 urine samples (by gas chromatography-mass spectrometry [GC-MS]: 88% of 64 urine samples) obtained from Rave attendees contained MDMA and/or 3,4 methylenedioxyamphetamine (MDA) alone or in combination with amphetamine, methamphetamine, or other designer amphetamines such as 3,4 methylenedioxyethylamphetamine (MDEA). This suggests that the majority of the Ravers are multidrug users. At the manufacturer's suggested cutoffs, the Abbott TDx Amphetamine/Methamphetamine II and the new Roche HS Amphetamine/MDMA assays demonstrated greater detection sensitivity for MDMA than the other amphetamine immunoassays tested (Abuscreen OnLine Hitachi AMPS, Abuscreen OnLine Integra AMPS, Abuscreen OnLine Integra AMPSX, CEDIA AMPS, and EMIT II AMPS). There is 100% agreement between each of the two immunoassays with the reference chromatographic methods, HPLC-DAD and GC-MS, for the detection of methylenedioxyamphetamines. PMID- 11386640 TI - 1H NMR spectroscopic investigation of serum and urine in a case of acute tetrahydrofuran poisoning. AB - This article reports the investigation by 1H nuclear magnetic resonance (1H NMR) spectroscopy of biological fluids in a case of intentional poisoning with tetrahydrofuran (THF). Occupational exposures to this solvent are well documented, but acute poisoning cases are extremely rare, and the one presented here is the second known case of this kind. Urine and serum samples were collected. Without any pretreatment, the presence of THF was confirmed by characteristic resonances at 1.90 and 3.76 ppm; high lactate levels were also observed. The presence of gamma-hydroxybutyric acid (GHB) was noted. Quantitative analysis was performed by relative integration of peak areas. THF concentrations were 813 and 850 mg/L (11.3 and 11.8 mmol/L), and GHB concentrations 239 and 2,977 mg/L (2.3 and 28.6 mmol/L) in serum and urine, respectively. A gas chromatographic-mass spectrometric method confirmed 1H NMR observations. The origin of GHB detected in serum and urine is also discussed. PMID- 11386641 TI - Simultaneous determination of chlorpyrifos, permethrin, and their metabolites in rat plasma and urine by high-performance liquid chromatography. AB - A high-performance liquid chromatographic (HPLC) method was developed for the separation and quantitation of the insecticide chlorpyrifos (O,O-diethyl-O[3,5,6 trichloro-2-pyridinyl] phosphorothioate), its metabolites chlorpyrifos-oxon (O,O diethyl-O[3,5,6-trichloro-2-pyridinyl] phosphate) and TCP (3,5,6-trichloro-2 pyridinol), the insecticide permethrin (3-(2,2-dichloro-ethenyl)-2,2 dimethylcyclopropanecarboxylic acid-(3-phenoxyphenyl) methylester), and two of its metabolites, m-phenoxybenzyl alcohol and m-phenoxybenzoic acid, in rat plasma and urine. The method is based on using C18 Sep-Pak cartridges for solid-phase extraction and reversed-phase HPLC. The compounds were separated using a gradient of 1 to 80% acetonitrile in water (pH 3.2) at a flow rate ranging between 1 and 1.5 mL/min in a period of 17 min and gradient UV detection ranging between 210 and 280 nm. The retention times ranged from 9.3 to 14.5 min. The limits of detection ranged between 20 and 150 ng/mL, whereas the limits of quantitation were 150-200 ng/mL. The respective average percentage recoveries of chlorpyrifos, chlorpyrifos-oxon, TCP, permethrin, m-phenoxybenzyl alcohol, and m-phenoxybenzoic were 78.6 +/- 6.4, 72.8 +/- 6.8, 84.8 +/- 8.0, 79.2 +/- 8.4, 80.5 +/- 7.2, and 82.3 +/- 7.1 from five spiked plasma samples and 77.5 +/- 8.1, 72.8 +/- 8.3, 79.9 +/- 6.4, 79.1 +/- 8.9, 80.5 +/- 7.6, and 81.4 +/- 7.8 from urine samples. The relationship between peak areas and concentration was linear for concentrations between 200 and 2,000 ng/mL. This method was used to analyze these chemicals and metabolites following dermal administration in rats. PMID- 11386642 TI - Clenbuterol in the horse: confirmation and quantitation of serum clenbuterol by LC-MS-MS after oral and intratracheal administration. AB - Clenbuterol is a beta2 agonist/antagonist bronchodilator, and its identification in post-race samples may lead to sanctions. The objective of this study was to develop a specific and highly sensitive serum quantitation method for clenbuterol that would allow effective regulatory control of this agent in horses. Therefore, clenbuterol-d9 was synthesized for use as an internal standard, an automated solid-phase extraction method was developed, and both were used in conjunction with a multiple reaction monitoring liquid chromatography-tandem mass spectrometry (LC-MS-MS) method to allow unequivocal identification and quantitation of clenbuterol in 2 mL of serum at concentrations as low as 10 pg/mL. Five horses were dosed with oral clenbuterol (0.8 microg/kg, BID) for 10 days, and serum was collected for 14 days thereafter. Serum clenbuterol showed mean trough concentrations of approximately 150 pg/mL. After the last dose on day 10, serum clenbuterol reached a peak of approximately 500 pg/mL and then declined with a half-life of approximately 7 h. Serum clenbuterol declined to 30 and 10 pg/mL at 48 and 72 h after dosing, respectively. By 96 h after dosing, the concentration was below 4 pg/mL, the limit of detection for this method. Compared with previous results obtained in parallel urinary experiments, the serum-based approach was more reliable and satisfactory for regulation of the use of clenbuterol. Clenbuterol (90 microg) was also administered intratracheally to five horses. Peak serum concentrations of approximately 230 pg/mL were detected 10 min after administration, dropping to approximately 50 pg/mL within 30 min and declining much more slowly thereafter. These observations suggest that intratracheal administration of clenbuterol shortly before race time can be detected with this serum test. Traditionally, equine drug testing has been dependent on urine testing because of the small volume of serum samples and the low concentrations of drugs found therein. Using LC-MS-MS testing, it is now possible to unequivocally identify and quantitate low concentrations (10 pg/mL) of drugs in serum. Based on the utility of this approach, the speed with which new tests can be developed, and the confidence with which the findings can be applied in the forensic situation, this approach offers considerable scientific and regulatory advantages over more traditional urine testing approaches. PMID- 11386643 TI - IOL use in children with uveitis-related cataract. PMID- 11386644 TI - Effectiveness of mitomycin C in reducing reformation of adhesions following surgery for restrictive strabismus. AB - PURPOSE: To evaluate the efficacy of mitomycin C in reducing reformation of adhesions following surgery for restrictive strabismus. METHODS: Sixteen patients with restrictive strabismus resulting from trauma, retinal detachment surgery, and multiple squint surgeries were randomized to either a mitomycin group (8 patients) or a control group (8 patients). Mitomycin C (0.2 mg/mL) was applied intraoperatively for 5 minutes after release of adhesions. Range of passive ductions was scored for comparison. Postoperative follow-up ranged from 3-18 months. RESULTS: Compared to controls, the mitomycin C group had better range of passive ductions. There were no complications related to the cornea, intraocular pressure, uveitis, or muscle disinsertion in the mitomycin C group. CONCLUSION: Intraoperative application of mitomycin C (0.2 mg/mL for 5 minutes) is a safe and effective adjunct to surgery in the treatment of restrictive strabismus. PMID- 11386645 TI - Echography of retinoblastoma: histopathologic correlation and serial evaluation after globe-conserving radiotherapy or chemotherapy. AB - PURPOSE: To assess the sensitivity of echography in detecting retinoblastoma, compare tumor features observed by echography with histopathology data, and assess the usefulness of echography in serially following retinoblastoma tumors after globe-conserving treatments. METHODS: The medical and echography records of all patients treated for retinoblastoma at the Bascom Palmer Eye Institute between 1991 and 1997 were reviewed. All eyes underwent pretreatment echographic evaluation, and eyes treated with external beam radiotherapy, brachytherapy, or chemotherapy underwent serial follow-up echography. RESULTS: Sixty-nine eyes of 48 patients were identified. Echography demonstrated evidence of retinoblastoma in 69 of 69 (100%) eyes and calcification in 63 (91.3%) eyes. Histopathology was superior to echography in detecting optic nerve invasion, extraocular extension, and presence of calcification. CONCLUSION: Echography is a useful adjunct to indirect ophthalmoscopy in establishing the diagnosis of retinoblastoma. While not as specific as histopathology, echographic evaluation before and after treatment of retinoblastoma permits monitoring of treatment response and may aid in detecting recurrent tumor growth or failure to respond to treatment. PMID- 11386646 TI - Effect of retinopathy of prematurity on resolution acuity development in 1- to 3 year-old children. AB - PURPOSE: To assess the pattern of visual development in children aged 1-3 years with stage 1-3 retinopathy of prematurity (ROP). METHODS: One hundred forty-four infants weighing <1500 g were divided into three groups according to ROP stage. Randomly chosen preterm infants were the controls. Ophthalmic examinations started 4-7 weeks after birth and were performed until the retina was fully vascularized or until any ROP that had developed resolved. Preferential looking acuity measurements were carried out at 12, 18, 24, and 36 months of age. RESULTS: Infants with stage 1-2 or no ROP showed evidence of improving acuity development after 12 months. Infants with stage 3 ROP had significantly lower acuity scores compared to infants with stage 1-2 or no ROP at the 18- and 24 month follow-up examinations (P<.001). At 36 months, the visual acuity of infants with stage 3 ROP showed evidence of improving development but still lagged behind infants with stage 1-2 or no ROP. CONCLUSION: Although mild ROP does not seem to have a considerable effect on the development of resolution acuity until age 3, severe forms of ROP may be associated with impaired visual development, which strongly suggests the necessity of periodic monitoring of early visual acuity in infants with ROP. PMID- 11386647 TI - Near-work activity and myopia in rural and urban schoolchildren in China. AB - PURPOSE: To examine the prevalence of myopia in rural and urban schoolchildren in Xiamen, China, and to assess the impact of environmental factors on rates of myopia. METHODS: Second-grade children attending either a city (n=119) or rural (n=91) school in Xiamen, China, were examined using cycloplegic autorefraction and biometry. Detailed questions on socioeconomic status, near-work activity, reading and writing habits, and family histories of myopia were asked in a face to-face interview using a standard questionnaire. RESULTS: The prevalence of myopia was 19.3% (95% confidence interval [CI], 12.3, 29) in the city and 6.6% (95% CI, 2.4, 14.3) in the countryside. The average hours per day children spent reading and writing outside of school was 2.2 hours in the city compared with 1.6 hours in the countryside (P<.0001). In both schools, the odds ratio for total reading and writing, adjusted for parental history of myopia, was 2.2 (95% CI, 1.1, 4). CONCLUSION: These data suggest the prevalence of myopia is higher in the city than in the countryside. One possible explanation for these different rates could be that schoolchildren in the city spend more time reading and writing outside of school compared with children in the countryside. Myopic children in both the city and the countryside spent more time reading and writing compared with nonmyopic children. This increased near-work activity may contribute to the prevalence of myopia. PMID- 11386648 TI - Ocular and clinical manifestations of Mobius' syndrome. AB - PURPOSE: To assess ocular and otorhinolaryngologic manifestations and intellectual ability in patients with Mobius' syndrome. METHODS: Patients with Mobius' syndrome underwent prospective ophthalmic, genetic-clinical, and otorhinolaryngologic examinations as well as psychological evaluation. RESULTS: Sixteen patients with Mobius' syndrome between the ages of 8 months and 10.6 years underwent ocular examination. Esotropia was present in 12 (75%) patients and V-pattern in 8 (50%). Limited abduction was present in 30 (93.8%) eyes, and limited adduction was present in 21 (65.6%) eyes. The most frequent refractive error was compound hyperopic astigmatism (13 [40.6%] eyes). Eleven (68.8%) patients had lagophthalmos and 12 (75%) patients had bilateral epicanthus. Unilateral amblyopia was present in 2 (12.5%) patients. Clubfoot was the most common lower limb defect (7 [43.8%] patients). Cranial nerve impairments included paralysis of 7th nerve in all patients, paralysis of the 12th nerve in 13 patients, and paralysis of the 9th and 10th nerves in 3 patients. Evaluation of intellectual ability showed that 4 (25%) patients had normal intelligence. The mothers of 3 (18.8%) patients used misoprostol during the first trimester of pregnancy. CONCLUSION: Prominent ophthalmic features of Mobius' syndrome in this series were esotropia, V-pattern, abduction limitation, and compound hyperopic astigmatism. Intellectual assessment showed some degree of mental retardation in 75% of patients. Mobius' syndrome is associated with prenatal exposure to misoprostol. PMID- 11386649 TI - Follow-up after probing for congenital nasolacrimal duct obstruction. AB - PURPOSE: To establish the optimum timing of follow-up after probing for uncomplicated congenital nasolacrimal duct obstruction. METHODS: A retrospective review of notes of all patients probed over a 4-year period was performed. Timing of symptom resolution was evaluated and compared to the results of a questionnaire sent to all parents in June 1996. RESULTS: Most patients were asymptomatic or improved by 3 months postoperatively, and the long-term success rate was 96.6%. CONCLUSION: Optimum timing of follow-up should be 3 months postoperatively, if necessary at all. PMID- 11386650 TI - Clinicopathologic factors related to metastasis in retinoblastoma. AB - PURPOSE: To investigate the orbital invasion and metastatic pattern in retinoblastoma, and to analyze the correlation with other pathologic and prognostic factors. METHODS: Pathologic and admission records of 45 patients with a pathologically confirmed diagnosis of retinoblastoma during the past 20 years were reviewed. Clinical risk factors and pathologic findings were analyzed for association with tumor invasion/metastasis using chi-square, Fisher's exact, and Student's t tests. RESULTS: Clinical risk factors such as exophthalmos, cataract, and pseudohypopyon were significantly correlated with tumor invasion/metastasis. The pathologic finding of choroid involvement was highly predictive of tumor invasion/metastasis. Delayed enucleation (>3 months) also was significantly correlated with tumor invasion/metastasis. CONCLUSION: Tumor invasion/metastasis is strongly associated with various ocular, pathologic, and treatment outcome factors. Identification of these risk factors may help plan treatment regimens. PMID- 11386651 TI - Herpes zoster ophthalmicus. PMID- 11386652 TI - Adrenal suppression and growth retardation from ocular corticosteroids. PMID- 11386653 TI - Primary levator synkinesis associated with eye movement. PMID- 11386654 TI - Nystagmus blockage syndrome associated with accommodative spasm. PMID- 11386655 TI - Spontaneous ophthalmic artery occlusion in a child. PMID- 11386656 TI - Determination of additives in food by capillary electrophoresis. AB - The application of capillary electrophoresis (CE) to the analysis of additives in food has been reviewed. Additives included in the review are preservatives, antioxidants, sweeteners, colourings, caffeine, niacin, choline, nitrate, and nitrite. The review highlights the versatility of CE in separating this often widely disparate group of compounds. The application of the methods to real food samples is also discussed. PMID- 11386657 TI - Separation of iota-, kappa and lambda-carrageenans by capillary electrophoresis. AB - The present study reports a novel method for the separation of the high-molecular weight anionic polysaccharides, iota, kappa, and lambda carrageenans, in capillary electrophoresis (CE). Carrageenan samples are first derivatised with 9 aminopyrene-1,4,6-trisulfonic acid (APTS), separated in an ammonium acetate background electrolyte (BGE) and detected with laser-induced fluorescence (LIF). The effects of changes of instrumental parameters (temperature, injection mode, field strength) and the composition of the BGE (concentration and pH) are reported, and are explained in terms of the physical chemistry of the BGE and the biopolymers. Optimal separation conditions for kappa, iota, and lambda carrageenans, including an APTS internal standard, were found in a polyvinyl alcohol coated capillary with an ammonium acetate BGE of low concentration (25 mM) and moderate pH (8.0). This BGE gave the best reproducibility in tests on iota/kappa mixtures, with relative standard deviations (RSDs) in migration times and normalised peak areas (relative to the APTS internal standard) of less than 0.1% and 1%, respectively. Using this BGE at 50 degrees C and a voltage of 30 kV, all three carrageenan subtypes were separated in a run time of 3 min. PMID- 11386658 TI - The application of capillary electrophoresis to the analysis of vitamins in food and beverages. AB - Capillary electrophoretic (CE) methods have been used to separate and determine a wide range of water-soluble vitamins in pharmaceutical preparations, but has found limited application in determining vitamins at naturally occurring levels in food and beverages. CE has been used to determine vitamin C in fruits and beverages, niacin in a range of foods and thiamine in samples of meat and milk. The CE methodologies used to determine vitamins in pharmaceutical preparations and biological fluids are also included as examples of the effectiveness of CE in vitamin analysis. PMID- 11386659 TI - Determination by capillary electrophoresis of total and available niacin in different development stage of raw and processed legumes: comparison with high performance liquid chromatography. AB - The available and total niacin evolution during maturation of yellow pea lupine (Lupinus luteus L., cv. Juno), pea (Pisum sativum L., cv. Ergo), faba bean (Vicia faba sp. minor Harz, cv. Tibo) and in germinated and high-pressure heated peas (Pisum sativum L., cv. Esla) have been determined by capillary electrophoresis (CE). The results have been compared with those obtained by high-performance liquid chromatography (HPLC). The values obtained by CE were similar to those obtained by HPLC. Maturation of seeds significantly reduced the available and total niacin content in legumes. The available/total niacin ratio during seed maturation depends on the type of legume. In faba beans and peas, a reduction was observed which was more pronounced in the case of peas. For lupine seeds, the ripening produced an increase (34%) in the available/total niacin ratio 50 days after flowering (DAF). Pea germination produced an overall increase in available and total niacin content although the available/total niacin ratio decreased. High pressure heating of pea yielded an increase in the available niacin content and available/total niacin ratio but the total niacin content did not change. PMID- 11386660 TI - Quantification of L-ascorbic acid and total ascorbic acid in fruits and spinach by capillary zone electrophoresis. AB - A standard curve for the quantification of L-ascorbic acid (L-AA) by capillary zone electrophoresis (CZE) was established, and the quantification of ascorbic acid and total ascorbic acid in fruits (lemon, Sunkist, and pineapple) and spinach were performed using D-isoascorbic acid (D-IAA) as an internal standard. The minimum detection limits (MDLs) for L-AA and D-IAA were determined to be 1 and 2 microg/mL, respectively, at 265 nm. Dehydroascorbic acid (DHAA) in fruits and spinach was quantified in the presence of DL-homocysteine. The recoveries for L-AA in these juices were between 95 and 105%. PMID- 11386661 TI - Capillary electrophoresis for the analysis of food proteins of animal origin. AB - In recent years, capillary electrophoresis (CE) has become an analytical technique with many applications in the study of food proteins and peptides. This review describes the existing CE methods of analysis of milk, egg, meat and fish proteins and peptides. The major developments in the application of CE to solve different problems in food technology, such as the assessment of technological processes, quality, and authenticity control of animal foods, are considered. A section dealing with future directions on the analysis of food proteins by CE is also included. PMID- 11386662 TI - Recent developments in high-performance capillary electrophoresis of cereal proteins. AB - Cereal proteins play important nutritional and functional roles in human foods and are also important components of animal feeds. As such, cereals are a major economic factor around the world. Because of their importance, cereal proteins have been widely studied. A new emerging technique for studying cereal proteins is high-performance capillary electrophoresis (HPCE). This review focuses mainly on new methods and applications of HPCE to cereal proteins that have been reported in the last three years. PMID- 11386663 TI - Qualitative and quantitative analysis of proteins and peptides in milk products by capillary electrophoresis. AB - Milk protein is an important component of the human diet throughout much of the world. The ability to assess the relative composition and integrity of milk proteins or peptides in dairy foods or food ingredients is important because these molecules have a profound effect on product functionality and quality. This communication describes two capillary electrophoretic methods that are useful for the analysis of proteins and casein-derived peptides in cheese and milk products. One technique, which uses a buffer containing citrate/phosphate (pH 3.3), 4 M urea, and a polymeric additive in a coated capillary, is useful for qualitative and quantitative analysis of proteins and peptides in milk, cheese, and whey products. The second method employs a citrate/phosphate buffer (pH 2.8) and a bare silica capillary, and is well suited for the analysis of small, casein derived peptides in aqueous cheese extracts. PMID- 11386664 TI - Novel approaches to the analysis of the Maillard reaction of proteins. AB - The Maillard reaction comprises a complex network of reactions which has proven to be of great importance in both food science and medicine. The majority of methods developed for studying the Maillard reaction in food have focused on model systems containing amino acids and monosaccharides. In this study, a number of electrophoretic techniques, including two-dimensional gel electrophoresis and capillary electrophoresis, are presented. These have been developed specifically for the analysis of the Maillard reaction of food proteins, and are giving important insights into this complex process. PMID- 11386665 TI - Requirements for the application of protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis and randomly amplified polymorphic DNA analyses to product speciation. AB - Raw, cooked, fried, smoked and gravad (brine-cured) products were analyzed by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of proteins and by randomly amplified polymorphic DNA (RAPD) in order to identify the species used in their manufacture. The discriminatory power of SDS-PAGE was dependent primarily on the composition and secondarily on the size of the gels: the Laemmli buffer system with 15% acrylamide and 0.087% piperazine diacrylamide separating gels resolved more discriminant protein bands than any of the commercial gels tested. Some of the processing conditions induced alterations in the protein patterns that made identification dubious. Differentiation even between closely related species was easier by RAPD than by SDS-PAGE. Neither the processing conditions nor the tissue from which the DNA was extracted had a significant effect on the RAPD profiles. For identifications based on SDS-PAGE, one should use an optimized gel composition and separate the sample under analysis in the same gel as the references. For RAPD-based identifications, the unknown sample should be amplified together with reference samples and separated in the same gel. PMID- 11386666 TI - Computer-assisted evaluation of isoelectric focusing patterns in electrophoretic gels: identification of smoothhounds (Mustelus mustelus, Mustelus asterias) and comparison with lower value shark species. AB - In this work, the most commercially important Selachian species, smoothhound (Mustelus mustelus) and starry smoothhound (Mustelus asterias), have been identified by polyacrylamide isoelectric focusing (IEF-PAGE), along with several shark species of minor commercial value. In Italy, these two smoothhound species are commonly subjected to fraudulent substitution with lesser valued sharks. After the electrophoretic runs, the band patterns of the two Mustelus species were compared with those of dogfish (Scyliorhinus canicula), spurdog (Squalus acanthias), blue shark (Prionace glauca) and black-mouthed dogfish (Galeus melanostomus), both visually and with gel analysis software. The actual isoelectric points were then submitted to cluster analysis to differentiate the single species, despite the possible occurrence of electrophoretic variations or protein polymorphism. Every shark showed species-specific band patterns and could therefore be well differentiated, as confirmed by statistical analysis. PMID- 11386667 TI - Protein changes in post mortem sea bass (Dicentrarchus labrax) muscle monitored by one- and two-dimensional gel electrophoresis. AB - This study was devoted to the identification of specific peptides and proteins which can be used as indicators of freshness in fish. The post mortem evolution of protein patterns in farmed sea bass muscle was monitored by Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE) after 0, 2, 4, and 6 days cold storage. SDS electrophoresis, of total proteins and proteins soluble in low-ionic-strength solutions, revealed the gradual disappearance of a protein band of 16 kDa immediately after fish death. 2-DE allowed the classification of fish samples according to post mortem time. Three spots of interest, which disappeared progressively, were identified on the 2-DE patterns. Further research is required to establish the identity of these polypeptides and to evaluate their expression and post mortem evolution in another fish species. PMID- 11386668 TI - Characterization and partial sequencing of species-specific sarcoplasmic polypeptides from commercial hake species by mass spectrometry following two dimensional electrophoresis. AB - The Merluccidae family comprises marine species, some of them of high commercial value and others less appreciated, whose commercialization in Europe under the generic name of "hake" is highly remarkable. The potential of proteomics was employed in this study with the aim of achieving the differential characterization of five different hake species: Merluccius merluccius (European hake), M. australis (Southern hake), M. hubbsi (Argentinian hake), M. gayi (Chilean hake), and M. capensis (Cape hake), some of them very closely related. Species-specific polypeptides were observed for the five hake species studied in isoelectric focusing (IEF) and/or two-dimensional electrophoresis (2-DE) high resolution gels. The peptide mass maps of two polypeptide groups, previously selected by 2-DE analysis as potentially species-specific, were obtained by "in gel" tryptic digestion, followed by matrix assisted laser desorption/ ionization time of flight-mass spectrometry (MALDI-TOF-MS). Analysis of group A polypeptides (with pI in the range of 5.0-5.5 and molecular mass of 17 kDa), allowed the differential classification of the hake species into two groups: the East Atlantic coast group and the West Atlantic coast group. Moreover, the peptide mass-maps from the heat-resistant parvalbumin fraction (pI below 4.5; molecular mass <12 kDa) allowed the detection of a peptide characteristic of M. australis not present in the other four hake species tested. A specific 17 kDa protein from M. merluccius was also partially sequenced by nanospray-ion trap-tandem MS, revealing a high homology with rat nucleoside diphosphate kinase A (NDKA). This work opens the way to the application of proteomics to the differential characterization of commercial hake species at the molecular level. PMID- 11386669 TI - Determination of phenolic constituents in citrus samples by on-line coupling of a flow system with capillary electrophoresis. AB - Phenolic compounds extracted from different citrus were determined. Calibration, extraction, elution, and introduction into the sample vial was carried out automatically by a continuous flow system (CFS) coupled to capillary electrophoresis (CE) equipment via a programmable arm. The only manual operation was the centrifugation of the sample to remove the pulp. The supernatant solutions were introduced into the CFS-CE system. A C-18 minicolumn coupled into the CFS was used to perform cleanup of the samples. The analytes were eluted from the minicolumn using methanol. Quantitative analysis was carried out by the standard addition method. The method presented allows a fast, quantitative, and reproducible determination of six main phenolic compounds in citrus samples, with precision in the range of 3.0-6.5%, expressed as relative standard deviations. PMID- 11386670 TI - Fast determination of procyanidins and other phenolic compounds in food samples by micellar electrokinetic chromatography using acidic buffers. AB - Procyanidins are phenolic oligomers, mainly composed of (+)-catechin and (-) epicatechin units that exhibit certain sensorial and physiological properties of interest (e.g., astringency and bitterness of food, antioxidant activity, etc.). This paper shows the development of a micellar electrokinetic chromatography (MEKC) method for the separation of three procyanidin dimers (B1, B2, and B3), their monomers ((+)-catechin and (-)-epicatechin), and the cis- and trans-forms of p-coumaric acid. Separation conditions are optimized in terms of buffer pH, SDS concentration, and washing routine between injections. The best results in terms of peak resolution and reproducibility between separations were obtained with a MEKC running buffer at pH 5 with 100 mM SDS and a washing routine that includes a rinse step with 0.1 M sodium hydroxide. Using this new MEKC method it is possible to separate in less than 5 min the seven substances. More interestingly, it is demonstrated that the low pH used in this MEKC method allows one to obtain clean electropherograms when samples are injected. The method is shown to be reproducible between different days with relative standard deviation (RSD) values lower than 1% for migration times and lower than 7% for peak areas (3 days, 24 injections). The usefulness of this procedure to determine these compounds in effluents from food processing (i.e., soaking water from lentils, white beans and black beans) and in food by-products (i.e., almond peels) considered as potential procyanidin sources is demonstrated. To our knowledge, this is the first report of separation and determination of procyanidins in food samples done by capillary electrophoresis. PMID- 11386671 TI - Analysis of phenolic compounds in Spanish Albrarino and Portuguese Alvarinho and Loureiro wines by capillary zone electrophoresis and high-performance liquid chromatography. AB - The concentration of different phenolic compounds was measured in Spanish Albarino and Portuguese Alvarinho and Loureiro white wines by capillary zone electrophoresis (CZE), in order to characterize them. Although all samples presented the same qualitative pattern (characterized by tyrosol; (-) epicatechin; syringic acid; ferulic acid; p-coumaric acid; caffeic acid, gallic acid; 3,4-dihydroxybenzoic acid; cis-coumaroyl tartaric acid (COUTA); trans COUTA; trans-caffeoyl tartaric acid (CAFTA), and hydroxycinnamic esters), some quantitative differences were observed. When samples were analyzed by high performance liquid chromatography (HPLC), in order to compare the results obtained by both techniques, no significant qualitative or quantitative differences were obtained. Nevertheless, CZE proved to be a more convenient technique for the routinary analyses of these wines, due to better separation of the different compounds, better peak shapes, and higher speed than HPLC. PMID- 11386672 TI - Application of micellar electrokinetic capillary chromatography for quantitative analysis of quercetin in plant materials. AB - A simple method for the routine determination of quercetin in biological samples was developed. The method consists of hydrolysis of bonded quercetin, its isolation and preconcentration on solid-phase extraction (SPE) column and a final analytical step using micellar electrokinetic capillary chromatography. The working range, linear range, the limit of quantification, and the measurement uncertainty were tested in validation. The method is suitable for quercetin determination in fruit and vegetable samples. PMID- 11386673 TI - Determination of free amino acids in infant food by capillary zone electrophoresis with mass spectrometric detection. AB - A method for the quantitative determination of free amino acids in infant food samples using capillary electrophoresis (CE) with electrospray-mass spectrometric (ESI-MS) detection is presented. According to the zwitterionic nature of the analytes, two different modes of separation as well as detection were tested; highly acidic carrier electrolytes combined with MS detection in the positive ion mode proved to be the optimum solution. Sensitivity as well as linearity of the method were sufficient to allow the analysis of all solutes of interest in a single run. In this way, free amino acids could be analyzed in a variety of infant food preparations without any sample pretreatment or derivatization step. PMID- 11386674 TI - Comparison of protocols for pulsed-field gel electrophoresis of Clostridia. AB - Genotyping of bacterial strains via pulsed-field gel electrophoresis has to be considered an important tool for epidemiological investigations in food hygiene as well as in other areas. Yet, a major disadvantage of this method is its long duration. Therefore, rapid procedures for DNA isolation and restriction are being sought. One such protocol was modified and further shortened to two days. This short protocol was used for macrorestriction analysis of 34 strains of 25 different Clostridium species. Parallel analyses were performed using a conventional 5-day protocol in order to compare the long and the short method by running the DNA samples obtained via both protocols on the same gel. In the case of nine strains, none of the two methods yielded satisfactory results, whereas for three strains the long protocol proved to be preferable to the short one. Comparable results were obtained using both methods in the case of 22 strains belonging to 17 different Clostridium species. PMID- 11386675 TI - Characterisation of carotenoid radical cations in liposomal environments: interaction with vitamin C. AB - Pulse radiolysis was used to generate the radical cations of beta-carotene and two xanthophylls, zeaxanthin and lutein, in unilamellar vesicles of dipalmitoylphosphatidyl choline. The rate constants for the reaction (repair) of these carotenoid radical cations with the water-soluble vitamin C were found to be similar (approximately 1x10(7) M(-1) s(-1)) for beta-carotene and zeaxanthin and somewhat lower (approximately 0.5x10(7) M(-1) s(-1)) for lutein. The results are discussed in terms of the microenvironment of the carotenoids and suggest that for beta-carotene, a hydrocarbon carotenoid, the radical cation is able to interact with a water-soluble species even though the parent hydrocarbon carotenoid is probably entirely in the non-polar region of the liposome. PMID- 11386676 TI - Gramicidin A and its complexes with Cs+ and Tl+ ions in organic solvents. A study by steady state and time resolved emission spectroscopy. AB - Gramicidin A (gr A), a linear pentadecapeptide containing four trp residues has been studied using steady state and time resolved fluorescence (at 298 K) and phosphorescence (at 77 K) in methanol (CH3OH), ethanol (C2H5OH), dimethyl sulfoxide (DMSO), 1,4-dioxane, 2-methyl tetrahydrofuran (2-MeTHF), ethanol/benzene (C2H5OH/C6H6) mixed solvent. Similar studies have also been carried out in CH3OH containing monovalent cations K+, Cs+, Tl+ and divalent cation Ca2+. Lambda(max) of fluorescence is found to be a good signature of the different forms having double helical structure [dh (1) to dh (4)] (J. Struct. Biol. 121 (1998) 123-141). Steady state and time resolved quenching studies of gr A by KI in CH3OH and DMSO and life time of the emitting singlet states of gr A support that gr A exists as a mixture of different forms of double helical (dh) structure [dh (1) to dh (4)] in CH3OH and as a random coil structure in DMSO. This study further indicates that emitting trp residue in DMSO is better shielded than that in CH3OH. Phosphorescence spectra of gr A at 77 K in CH3OH glass suggests that gr A retains a particular conformation dh (3) in this matrix. The phosphorescence spectra of gr A [conformation dh (4)] in 2-MeTHF at 77 K is further red shifted indicating that among all the dh forms, dh (4) has the emitting trp residue in most hydrophobic environment. The hydrophobicity of the emitting tryptophan environment is thus found to be in the order: dh (1)1% . Albendazole's unique broad-spectrum activity is exemplified in the overall cure rates calculated from studies employing the recommended doses for hookworm (78% in 68 studies: 92%, for A. duodenale in 23 studies and 75% for N. americanus in 30 studies), A. lumbricoides (95% in 64 studies), T. trichiura (48% in 57 studies), E. vermicularis (98% in 27 studies), S. stercoralis (62% in 19 studies), H. nana (68% in 11 studies), and Taenia spp. (85% in 7 studies). The facts that albendazole is safe and easy to administer, both in treatment of individuals and in treatment of whole communities where it has been given by paramedical and nonmedical personnel, have enabled its use to improve general community health, including the improved nutrition and development of children. PMID- 11386685 TI - Ivermectin: effectiveness in lymphatic filariasis. AB - This detailed review of the published studies underlying ivermectin's recent registration for use in lymphatic filariasis (LF) demonstrates the drug's single dose efficacy (over the range of 20-400 microg/kg) in clearing microfilaraemia associated with both Wuchereria bancrofti and Brugia malayi infections of humans. While doses as low as 20 microg/kg could effect transient microfilarial (mf) clearance, higher dosages induced greater and more sustained mf reduction. The single dose of 400 microg/kg yielded maximal responses, but a number of practical considerations suggest that either 400 microg/kg or 200 microg/kg doses would be acceptable for use in LF control programmes. Associated safety assessments indicate that adverse events, which occur commonly following treatment of microfilaraemic individuals, develop not because of drug toxicity but because of host inflammatory responses to dying microfilariae killed by the ivermectin treatment. Ivermectin is, therefore, a highly effective and generally well tolerated microfilaricide that may soon become an essential component of many public health initiatives to interrupt transmission of lymphatic filarial infection in an effort to eliminate LF globally. PMID- 11386686 TI - An analysis of the safety of the single dose, two drug regimens used in programmes to eliminate lymphatic filariasis. AB - This review of the safety of the co-administration regimens to be used in programmes to eliminate lymphatic filariasis (albendazole + ivermectin or albendazole + diethylcarbamazine [DEC]) is based on 17 studies conducted in Sri Lanka, India, Haiti, Ghana, Tanzania, Kenya, Ecuador, the Philippines, Gabon, Papua New Guinea, and Bangladesh. The total data set comprises 90,635 subject exposures and includes individuals of all ages and both genders. Results are presented for hospital-based studies, laboratory studies, active surveillance of microfilaria-positive and microfilaria-negative individuals, and passive monitoring in both community-based studies and mass treatment programmes of individuals treated with albendazole (n = 1538), ivermectin (9822), DEC (576), albendazole + ivermectin (7470), albendazole + DEC (69,020), or placebo (1144). The most rigorous monitoring, which includes haematological and biochemical laboratory parameters pre- and post-treatment, provides no evidence that consistent changes are induced by any treatment; the majority of abnormalities appear to be sporadic, and the addition of albendazole to either ivermectin or DEC does not increase the frequency of abnormalities. Both DEC and ivermectin show, as expected, an adverse event profile compatible with the destruction of microfilariae. The addition of albendazole to either single-drug treatment regimen does not appear to increase the frequency or intensity of events seen with these microfilaricidal drugs when used alone. Direct observations indicated that the level of adverse events, both frequency and intensity, was correlated with the level of microfilaraemia. In non microfilaraemic individuals, who form 80-90% of the 'at risk' populations to be treated in most national public health programmes to eliminate lymphatic filariasis (LF), the event profile with the compounds alone or in combination does not differ significantly from that of placebo. Data on the use of ivermectin + albendazole in areas either of double infection (onchocerciasis and LF), or of loiais (with or without concurrent LF) are still inadequate and further studies are needed. Additional data are also recommended for populations infected with Brugia malayi, since most data thus far derive from populations infected with Wuchereria bancrofti. PMID- 11386687 TI - Salt fortified with diethylcarbamazine (DEC) as an effective intervention for lymphatic filariasis, with lessons learned from salt iodization programmes. AB - DEC-fortified salt has been used successfully as a principal public health tool to eliminate lymphatic filariasis (LF) in China and, less extensively, in several other countries. Studies from 1967 to the present conducted in Brazil, Japan, Tanzania, India, China, and Taiwan involving administration of DEC salt for 18 days to 1 year, have shown this intervention to be effective for both bancroftian and brugian filariasis, as measured by reductions in both microfilarial density and positivity, and in some studies through reduction in mosquito positivity rates as well. Furthermore, studies suggest specific advantages from using DEC salt, including lack of side effects, particularly for bancroftian filariasis, and ability to reduce prevalence below 1% when used in conjunction with standard regimens of DEC tablets. However, use of DEC salt as a control tool suffers from a concern that health authorities might find it difficult to manage a programme involving a commodity such as salt. In the past decade, the very successful global efforts to eliminate iodine deficiency through universal salt iodization have demonstrated that partnership with the salt industry can be both successful and effective as a public health tool. Use of DEC salt can be most successfully implemented in areas in which (a) there is adequate governmental support for its use and for elimination of filariasis, (b) filariasis-endemic areas are clearly defined, (c) political leaders, health officials and the salt industry agree that DEC salt is an appropriate intervention, (d) the salt industry is well-organized and has known distribution patterns, (e) a successful national salt iodization effort exists, (f) a monitoring system exists that ensures adequacy of salt iodine content during production and that can also measure household coverage, and (g) measurement of impact on transmission of LF with the new antigen or filarial DNA detection methods can be established. There are advantages and disadvantages of using DEC-fortified salt compared with other interventions for LF elimination programmes, but rather than being considered as a 'competing' intervention, DEC salt should be seen as an additional option. Indeed, it is likely that many countries will derive maximal benefit from the synergistic effects of combining different intervention strategies in their national programmes to eliminate lymphatic filariasis. PMID- 11386688 TI - Malnutrition and parasitic helminth infections. AB - The Global Burden of Disease caused by the 3 major intestinal nematodes is an estimated 22.1 million disability-adjusted life-years (DALYs) lost for hookworm, 10.5 million for Ascaris lumbricoides, 6.4 million for Trichuris trichiura, and 39.0 million for the three infections combined (as compared with malaria at 35.7 million) (World Bank, 1993; Chan et al. 1994); these figures illustrate why some scarce health care resources must be used for their control. Strongyloides stercoralis is the fourth most important intestinal worm infection; its nutritional implications are discussed, and the fact that its geographic distribution needs further study is emphasized. Mechanisms underlying the malnutrition induced by intestinal helminths are described. Anorexia, which can decrease intake of all nutrients in tropical populations on marginal diets, is likely to be the most important in terms of magnitude and the probable major mechanism by which intestinal nematodes inhibit growth and development. We present a revised and expanded conceptual framework for how parasites cause/aggravate malnutrition and retard development in endemic areas. Specific negative effects that a wide variety of parasites may have on gastrointestinal physiology are presented. The synergism between Trichuris and Campylobacter, intestinal inflammation and growth failure, and new studies showing that hookworm inhibits growth and promotes anaemia in preschool (as well as school-age) children are presented. We conclude by presenting rationales and evidence to justify ensuring the widest possible coverage for preschool-age children and girls and women of childbearing age in intestinal parasite control programmes, in order to prevent morbidity and mortality in general and specifically to help decrease the vicious intergenerational cycle of growth failure (of low-birth weight/intrauterine growth retardation and stunting) that entraps infants, children and girls and women of reproductive age in developing areas. PMID- 11386689 TI - Introduction: Opportunities to work together: intestinal helminth control and programmes to eliminate lymphatic filariasis. PMID- 11386690 TI - The public health importance of hookworm disease. AB - The public health significance of hookworm disease is briefly reviewed. The latest evidence confirms that blood loss caused by the worms' feeding activity in the gut is a contributing factor in the development of poor iron status leading to iron-deficiency anaemia. The World Health Organization has identified adolescent girls and women of child-bearing age as high-risk groups regarding the impact of hookworm disease. The merits of treating pregnant women with anthelminthic drugs after the first trimester are discussed. PMID- 11386691 TI - Global malnutrition. AB - The four most important forms of malnutrition worldwide (protein-energy malnutrition, iron deficiency and anaemias (IDA), vitamin A deficiency (VAD), and iodine deficiency disorders (IDD)) are examined below in terms of their global and regional prevalences, the age and gender groups most affected, their clinical and public health consequences, and, especially, the recent progress in country and regional quantitation and control. Zinc deficiency, with its accompanying diminished host resistance and increased susceptibility to infections, is also reviewed. WHO estimates that malnutrition (underweight) was associated with over half of all child deaths in developing countries in 1995. The prevalence of stunting in developing countries is expected to decline from 36% in 1995 to 32.5% in 2000; the numbers of children affected (excluding China) are expected to decrease from 196.59 millions to 181.92 millions. Stunting affects 48% of children in South Central Asia, 48% in Eastern Africa, 38% in South Eastern Asia, and 13-24% in Latin America. IDA affects about 43% of women and 34% of men in developing countries and usually is most serious in pregnant women and children, though non-pregnant women, the elderly, and men in hookworm-endemic areas also comprise groups at risk. Clinical VAD affects at least 2.80 million preschool children in over 60 countries, and subclinical VAD is considered a problem for at least 251 millions; school-age children and pregnant women are also affected. Globally about 740 million people are affected by goitre, and over two billions are considered at risk of IDD. However, mandatory salt iodisation in the last decade in many regions has decreased dramatically the percentage of the population at risk. Two recent major advances in understanding the global importance of malnutrition are (1) the data of 53 countries that links protein energy malnutrition (assessed by underweight) directly to increased child mortality rates, and (2) the outcome in 6 of 8 large vitamin A supplementation trials showing decreases of 20-50% in child mortality. PMID- 11386692 TI - The public health importance of Ascaris lumbricoides. AB - Numerous studies have shown that anthelminthic treatment can be effective in improving growth rates when given to malnourished children with ascariasis. Recent investigations have also indicated that Ascaris infections can affect mental processing in some school children. Poor socio-economic conditions are among the key factors linked with higher prevalences of ascariasis, as are defaecation practices, geophagia, cultural differences relating to personal and food hygiene, occupational necessity, agricultural factors, housing style, social class and gender. Chemotherapy is currently the major tool used for the strategic control of ascariasis as a short-term goal. In the long term, improvements in hygiene and sanitation are thought to aid long-term control considerably. Targeted treatment, especially when aimed at schoolchildren, has been a major focus of recent control efforts in some areas. Universal treatment reaches more people and thus decreases further aggregate morbidity, especially in nutritionally vulnerable preschool-age children. Selective treatment requires technical effort to identify heavily infected individuals; acceptance by the community may vary in less educated populations when some individuals receive treatment and others do not. Child-targeted treatment may be more cost-effective than population treatment in reducing the number of disease cases and, in high transmission areas, expanding coverage of a population can be a more cost effective strategy than increasing the frequency of treatment. PMID- 11386693 TI - The public health significance of Trichuris trichiura. AB - An estimated 1049 million persons harbour T. trichiura, including 114 million preschool-age children and 233 million school-age children. The prevalence of T. trichiura is high and may reach 95% in children in many parts of the world where protein energy malnutrition and anaemias are also prevalent and access to medical care and educational opportunities is often limited. The Trichuris dysentery syndrome (TDS) associated with heavy T. trichiura, which includes chronic dysentery, rectal prolapse, anaemia, poor growth, and clubbing of the fingers constitutes an important public health problem, as do lighter but still heavy infections, even if not strictly TDS, especially in children. The profound growth stunting in TDS can be reversed by repeated treatment for the infection and, initially, oral iron. However findings from Jamaica strongly suggest that the significant developmental and cognitive deficits seen are unlikely to disappear without increasing the positive psychological stimulation in the child's environment. The severe stunting in TDS now appears likely to be a reaction at least in part to a chronic inflammatory response and concomitant decreases in plasma insulin-like growth factor-1 (IGF-1), increases in tumor necrosis factor alpha (TNF-alpha) in the lamina propria of the colonic mucosa and peripheral blood (which likely decrease appetite and intake of all nutrients) and a decrease in collagen synthesis. Improvements in cognitive performance have been found after treatment for relatively heavy infections (without chronic dysentery) in school-going children; it is unclear precisely how much T. trichiura interferes with children's ability to access educational opportunities, but treatment of infections whenever possible is obviously sensible. The blood loss that can occur in T. trichiura infection is likely to contribute to anaemia, particularly if the child also harbours hookworm, malaria and/or has a low intake of dietary iron. Community control is important, particularly for the individuals within a population who harbour heavy worm burdens; this means children, with special attention to girls who will experience increased iron requirements and blood loss due to menstruation, pregnancies, and lactation. Mebendazole and albendazole, both of which are on the WHO Essential Drugs List, are very effective against T. trichiura; multiple doses are needed to attain complete parasitological cure in all cases. However the goal of control programmes in endemic areas is morbidity reduction, which follows when intensity of infection is significantly reduced. PMID- 11386694 TI - Animal models of intestinal nematode infections of humans. AB - In this paper we discuss several established and potential animal models for human parasitic infection, with a focus on rodent, pig and primate models and the nematodes Ascaris, Trichuris and Toxocara spp. Firstly, we discuss the relevance of choosing a suitable animal host to fit the particular study hypothesis, and the interaction between mathematical modelling and animal models. Secondly, we review the use of animal models for the study of nutrition-parasite interaction, evaluation of treatment and control strategies, and bacteria-parasite interactions. We show that rodent, pig and primate models are all very useful in parasitological research, and that each model has its limitations. However, based on recent experience with the pig-Ascaris and pig-Trichuris models, a more extensive use of the pig-parasite model is advocated, especially for the study of the interaction between human malnutrition and helminth infection, and congenital helminth infection. PMID- 11386695 TI - Language acquisition and language breakdown. PMID- 11386696 TI - Verb inflection and verb diversity in three populations: agrammatic speakers, normally developing children, and children with specific language impairment (SLI). AB - The present study focuses on the relation between a grammatical and a lexical semantic aspect of verb production. The spontaneous speech of three different populations (normally developing children, agrammatic aphasics, and children with a specific language impairment) has been analyzed with respect to the proportion of finite clauses and the diversity of the produced lexical verbs. The group results show that in the three populations both the proportion of finite verbs and the variability of the lexical verbs is low. When the individual scores are considered, differences between the normally developing children and the language impaired subjects show up. Whereas in normally developing children verb finiteness and verb variability go hand-in-hand, the reverse relationship between these variables is observed in the agrammatic aphasics and the children with a specific language impairment. Given this reverse relationship, it is probable to assume two separate disorders. We therefore suggest an impairment at the interface level where lexical information and syntactic structure are integrated during sentence production. PMID- 11386697 TI - The italian determiner system in normal acquisition, specific language impairment, and childhood aphasia. AB - The paper presents a comparison of the development of the Italian determiner system in three different populations: normally developing children, a child recovering from childhood aphasia from the age of 3 years, 9 months, and 11 specific language impairment (SLI) children. Data from Italian normal children provide evidence for the hypothesis (1) that no prefunctional stage exists as far as the determiner system is concerned and (2) that the syntactic properties of determiners play an essential triggering role early on. The analysis of the determiner system in the aphasic child has a double interest. On the one hand, it may help to shed light on some of the intriguing questions concerning this type of disorder; on the other, it may be relevant for the discussion of the notion of agrammatism. Results of the morphosyntactic analysis reveal that, apart from timing differences, recovery from childhood aphasia shares important features with normal development. Differently from mean length of utterance (MLU)-matched normal controls and the aphasic child, SLI children omit determiners significantly more often than almost any other functional category or free morpheme. We will argue that the reasons for the SLI children's atypical behavior have to be sought in the nonaccessibility to or in the misappreciation of one fundamental syntactic property of determiners: their role as elements that assign argumenthood to nominal expressions (Szabolcsi, 1987; Longobardi, 1994). PMID- 11386698 TI - Grammatism. AB - Findings from the literature on language development, dyslexia, and adult sentence processing provide a vehicle for comparing two models of the symptom complex associated with agrammatism. One model contends that agrammatism represents a deficit in linguistic structures. The other model maintains that the linguistic behavior associated with agrammatism is the result of a limitation in language processing. To adjudicate between the models, the present paper examines one linguistic construction, the restrictive relative clause. The results of experimental investigations across several subject populations reveal parallel patterns of linguistic behavior on this construction. The findings favor the processing limitation account of the linguistic difficulties experienced by agrammatic aphasics in comprehending sentences with a restrictive relative clause. PMID- 11386699 TI - The acquisition of complex predicates in japanese specifically language-impaired and normally developing children. AB - Japanese exhibits two different types of morphological processes. Some morphologically complex predicates are generated within the domain of the lexicon, whereas others are generated outside the domain of the lexicon. An elicited production task involving both types of complex predicates was administered to six Japanese children with specific language impairment (JSLI) and six children with normal language development (JNLD). The JSLI children experienced significant difficulty forming the lexicon-external complex predicates but much less difficulty with the lexicon-internal complex predicates while the performance of the JNLD children exhibited no such asymmetry. These preliminary results suggest that the deficit of SLI affects the ability to construct implicit procedural rules for morphology that are generated outside the lexicon while their lexical operations for morphology that are generated within the domain of the lexicon remain relatively unimpaired. PMID- 11386700 TI - Functional categories and syntactic operations in (Ab)normal language acquisition. AB - This article argues in favor of the hypothesis that computational complexity determines order of acquisition of functional categories by normal children and patterns of impairment vs. relative preservation of these categories in children with Specific Language Impairment. Complexity is defined in terms of the properties of functional categories of the language the (ab)normal child is acquiring. The results of a study on present/past tense production and comprehension, carried out with children with and without Specific Language Impairment confirm the predictions of the computational complexity hypothesis. It is shown that it is easier to compute kernel functional categories than those that introduce further semantic modifications in the sentence. PMID- 11386701 TI - Does agrammatic speech constitute a regression to child language? A three-way comparison between agrammatic, child, and normal ellipsis. AB - When children are in the process of learning their mother tongue, they show frequent use of nonfinite clauses, even though they produce finite clauses at the same time, thereby demonstrating the availability of the functional domain associated with finiteness. In this study the hypothesis was tested that this behavior results from an overuse of the normal elliptical repertoire that has also been observed in agrammatic aphasia. The purpose of this overuse is prevention of computational overload. In support of the hypothesis it was found that children behaved very similar to aphasics and normal adults with respect to the following parameters: (a) distribution of types of ellipsis, (b) elaboration of ellipses, (c) word order, (d) subject omission, (e) frequency of weak subject pronouns, and (f) verb type (eventivity). The results also support the Jackson/Jakobson regression hypothesis, at least at the grammatical level. PMID- 11386702 TI - Controversies about CP: a comparison of language acquisition and language impairments in Broca's aphasia. AB - In both language acquisition research and the study of language impairments in Broca's aphasia there is an ongoing debate whether or not phrase-structure representations contain the Complementizer Phrase (CP) layer. To shed some light on this debate, I will provide data on German child language and on German agrammatic Broca's aphasia. Analyses of subordinate clauses, wh-questions, and verb placement indicate that early child grammars do not generate the CP layer yet, whereas the ability to project the CP layer is retained in agrammatism. PMID- 11386703 TI - The vulnerable C-domain. AB - The purpose of this article is to document the existence of a group of natural language speakers who display non-target-like production of the syntax at the highest structural level, the C-domain, but produce the syntax of lower structural levels in a target-like way. This group consists of very early L1 learners, children with Specific Language Impairment, adult L2 learners, and patients with Broca's aphasia. The group is established on the basis of data from Swedish and German, but can presumably be discerned for any language. The assumption that the non-target-like production of C-domain syntax is related to Broca's area is discussed, as are some consequences for modern syntactic theory. PMID- 11386704 TI - Language disorders as a window on universal grammar: an abstract theory of agreement for IP, DP, and V-PP. AB - A new concept of Agreement (AGR) has been represented as a Formal Feature that can appear in a wide range of different configurations (Chomsky, 1998). A case study from language disorders supports and extends this abstract concept. The child shows no agreement in Inflectional Phrase me can and Determiner Phrase them eyes. We then extend the notion of AGR to include verb-Prepositional Phrase relations, where the child also systematically avoids certain prepositions (go beach). The analysis is supported by intuitional data from compounds (sweep with broom --> broom-swept). We also define a systematic notion of Possible deficit as a premature fixation of functional items which normally require additional Phi features. The notion of Maximization of Formal Features then emerges as a significant feature of learnability from both a normal and disordered perspective. PMID- 11386705 TI - Root nonfinite and finite utterances in child language and agrammatic speech. AB - Root nonfinite structures are frequent in both child language and agrammatic speech. Moreover, root nonfinites seem to have quite similar properties in both types of speech. Although many accounts are available for child language this is not the case for agrammatic speech. I present data of a Dutch agrammatic patient in this paper. The properties of her root nonfinites are best accounted for by the assumption of an underspecified Tense Phrase licensed by an empty element. In her finite sentences a dummy Tense element shows up. The use of either an empty or a dummy Tense element is a way of reducing the processing load of the utterance. The speech of this patient shows a dissociation between the verbal and the nominal domain. PMID- 11386706 TI - The early acquisition of verb meaning in German by normally developing and language impaired children. AB - The research reported here focuses on the early acquisition of event structure in German. Based on longitudinal studies from 5 normally developing (ND) and 6 language-impaired (LI) children, a model of "event structural bootstrapping" is presented that spells out how ND children log into the verb lexicon. They project a target-consistent event tree, depicting the head-of-event of transitions. Young LI children, failing to employ this bootstrapping strategy, resort to radically underspecified event representations. The results from a truth-value judgment experiment with 16 ND and 16 LI children showed that ND children perform correctly on transitional verbs, while LI children perform at chance level on the same tasks. These findings are accounted for by the model of event structural bootstrapping to the extent that LI children lack an explicit representation of the head-of-event. PMID- 11386707 TI - Comprehension of reversible relative clauses in specifically language impaired and normally developing Greek children. AB - This paper aims to investigate the syntactic comprehension of reversible relative clauses in a group of eight Greek children with specific language impairment (SLI) and two control groups of normally developing children matched on chronological and language age, respectively. An experiment using an acting out procedure was undertaken. Group analysis revealed that SLI children's performance is qualitatively different than that of both control groups. Interpreting the data, processes involved in syntactic comprehension are taken into consideration. It is claimed that processing demands impede SLI children's performance due to a deficit in their competence grammar. PMID- 11386708 TI - LF-interpretability and language development: a study of verbal and nominal features in Greek normally developing and SLI children. AB - This paper includes (i) a comparison of the developmental pattern of certain morphosyntactic features in normally developing (ND) Greek children with similar data from a group of children with specific language impairment and (ii) a new analysis of the differences found. The analysis is based on a minimalist notion of LF interpretability. Depending on the feature-specification of lexical items (i.e., [+/- interpretable] at LF), a different route of development follows. In addition, phonological salience of both interpretable and noninterpretable features is argued to play an important role in an account of crosslinguistic differences in both normal and exceptional development. PMID- 11386709 TI - Verb movement in acquisition and aphasia: same problem, different solutions evidence from Dutch. AB - This paper focuses on verb movement in agrammatism and child language. We present data from a sentence completion experiment with 6 Dutch agrammatic aphasics and 21 Dutch-speaking children. The experiment compares completion of matrix clauses (which require verb movement) and embedded clauses (where such movement is not required) in these two populations. The results reveal a clear asymmetry: Both agrammatics and children do very well with embedded clauses but fail in 50% with the matrix clauses. It is concluded that the problem which both populations are facing is one of verb movement rather than verb inflection. An error analysis of the responses reveals that, although both agrammatics and children try to avoid movement, they apply different strategies to achieve this goal. PMID- 11386711 TI - Ecotoxicity tests for compost applications. AB - Interest in the ecological effects of composting has been growing recently. However, no established methods are available for testing the toxicity of composted materials. Despite this, international and national quality requirements define that compost shall not contain any environmentally harmful substances. Safety requirements have to be fulfilled if the produced compost is intended for agricultural use. This literature review focuses on methods that could potentially be used to evaluate the ecotoxicity of compost. The toxicity test methods discussed are those employing microbes, enzymes, soil fauna, and plants. PMID- 11386712 TI - A critical review of the scientific literature on potential endocrine-mediated effects in fish and wildlife. AB - A rigorous approach to evaluating the reliability and relevance of experimental methods and results is critical for making appropriate, scientifically sound decisions. A literature evaluation framework was modified and adapted based on criteria described by Klimisch et al. (H. J. Klimisch, M. Andrae, and U. Tillman, 1997, Reg. Tox. Pharm. 25, 1-5) and requirements of the USEPA High Product Volume Challenge Program. This simple framework was used to evaluate an appropriate selection of peer-reviewed references to assign a Study Reliability "score" to the study design, endpoints, and conclusions compared to established data quality guidelines. Subsequently, the interpretation of the data was evaluated and assigned a Relevance Index based on the overall strength of evidence of endocrine modulation potential, which was evaluated based on the Weybridge (European Commission (1996). European workshop on the impact of endocrine disrupters on human health and wildlife. Report of proceedings from a workshop held in Weybridge, UK, 2-4 December 1996. Report reference EUR 17549, European Commission, DGXII, Brussels, Belgium.) definition. This report describes the evaluation of 50 peer-reviewed primary publications. The vast majority of studies reviewed were published in the period from 1996 to the present and most examined in vivo responses of fish to natural hormones, organochlorines, or alkylphenolic compounds. Of the studies reviewed, approximately 40% received a score of "2 Reliable with Restrictions," while approximately 60% received a score of "3-Not Reliable," where reliability was interpreted to include scientific integrity, technical credibility, endpoint relevance, and regulatory compliance. The most common deficiencies were inadequate methods, lack of suitable controls, exceeding aqueous solubility, inappropriate statistics, and unsupported conclusions. Based on the Relevance Index, few of the studies examined attained the level of quality necessary to identify the study results as providing an acceptable basis for evaluation of endocrine modulation potential directly resulting from identified deficiencies in meeting the established study reliability criteria. PMID- 11386713 TI - Ecotoxicology of glutaraldehyde: review of environmental fate and effects studies. AB - Glutaraldehyde is a biocide used in many industrial applications with potential releases to the environment. This review discusses the environmental fate and effects data on this important biocide. Information drawn from this review indicates that glutaraldehyde is acutely toxic to aquatic organisms. Glutaraldehyde is equally toxic to warm water and cold water fish, but is slightly more toxic to freshwater fish than salt water fish. The acute toxicity of glutaraldehyde for avian species is comparable to that for mammalian species. The toxicity of glutaraldehyde is not appreciably increased with repeated long term exposures. Results from environmental partitioning studies indicate that glutaraldehyde tends to remain in the aquatic compartment and has little tendency to bioaccumulate. Aqueous solutions of glutaraldehyde are stable at room temperature under acidic to neutral conditions, and to sunlight, but unstable at elevated temperatures, and under alkaline conditions. Glutaraldehyde is readily biodegradable in the freshwater environment and has the potential to biodegrade in the marine environment. Aquatic metabolism studies suggest that glutaraldehyde, under aerobic conditions, is metabolized to CO(2) via glutaric acid as an intermediate. Under anaerobic conditions, glutaraldehyde is metabolized to 1,5-pentanediol. Pretreatment with sodium bisulfite is the best method for inactivating glutaraldehyde prior to disposal to treatment systems. PMID- 11386714 TI - Structure of microbial communities in activated sludge: potential implications for assessing the biodegradability of chemicals. AB - Various methods used to assess the biodegradability of chemicals often employ activated sludge as an inoculum since chemicals that ultimately enter the environment are often discharged through wastewater. Differences in the structure and function of activated sludge microbial communities that may complicate interpretation of biodegradation tests could arise from differences in wastewater composition, wastewater treatment plant (WWTP) operation, or manipulations done after collection of the activated sludge. In this study, various methods were used to characterize the structure of microbial communities found in freshly collected activated sludge from WWTPs in Japan, Europe, and the United States, as well as sludge that had been continuously fed either sewage or a glucose-peptone mixture for several weeks after collection. Comparisons of biomass levels, whole community substrate utilization (determined using Biolog GN and GP plates), and phospholipid fatty acid (PLFA) profiles indicated there were both geographical and temporal differences among freshly collected activated sludge samples. Moreover, marked shifts in the structure of activated sludge microbial communities occurred upon continuous cultivation in the laboratory for 5 weeks using a glucose-peptone feed. These shifts were evident from whole-community substrate utilization and PLFA profiles as well as differences in the profiles of 16S rDNA genes from numerically dominant populations obtained by denaturing gradient gel electrophoresis and terminal restriction fragment analyses. Further studies are needed to better define the variability within and between activated sludge from wastewater treatment plants and laboratory reactors and to assess the impact of such differences on the outcome of biodegradability tests. PMID- 11386715 TI - Toxicity of copper to the collembolan Folsomia fimetaria in relation to the age of soil contamination. AB - The toxicity of copper to the collembolan Folsomia fimetaria L. was studied in soil incubated with copper sulfate for different periods before the introduction of collembolans, to assess the effect of aging of contamination on the toxicity of copper. Adult survival, reproduction, and juvenile size were assessed. No clear influence of differences in contamination age was detected. The data were compared with results from a study performed in soil sampled at an old copper contaminated site. Large differences in effects existed between spiked soil and field soil when concentrations were expressed on the basis of total soil copper concentrations. EC(10) and EC(50) values for reproduction in spiked soil were ca. 700 and 1400 mg Cu/kg soil, whereas no effects were found in field soil at copper concentrations up to 2500 mg/kg. Most of the differences disappeared when effects were expressed as a function of 0.01 M CaCl(2)-extractable soil copper. the lack of effects in field soil could be explained from the fact that in this field soil the CaCl(2)-extractable concentration was never higher than one-third of the EC(50) estimated for tests in the laboratory spiked soils. PMID- 11386716 TI - Metallothionein as bioindicator of freshwater metal pollution: European eel and brown trout. AB - The aim of this work was to evaluate the potential of metallothionein (MT) as a bioindicator of heavy metal pollution in brown trout and European eel in the field situation. River Ferrerias (North Spain) provided a good gradient of metal contamination: concentrations of heavy metals were elevated both in water and in sediments at the downstream (polluted) site and were low at the upstream (unpolluted) site. MT levels of brown trout exhibited statistically significant differences between sites. Although European eel at the polluted site had a higher MT content, differences were not significant. It is concluded that MT is a good bioindicator of heavy metal pollution in brown trout. PMID- 11386717 TI - Biotransformation, endocrine, and genetic responses of Anguilla anguilla L. to petroleum distillate products and environmentally contaminated waters. AB - The European eel (Anguilla anguilla L.) was exposed to diesel oil water-soluble fraction (DWSF) and gasoline water-soluble fraction (GWSF). The potential of these fractions to induce endocrine disruption, carbohydrate, and xenobiotic metabolism effects, as well as genotoxic responses, was investigated in a time course laboratory study (3 h to 6 days). Both water-soluble fractions induced a time-related increase in liver ethoxyresorufin-O-deethylase (EROD) activity, as well as the appearance of erythrocytic nuclear abnormalities (ENA) after a 6-day exposure, revealing its genotoxic properties. Initially, DWSF exposure revealed an inhibition of the typical stress responses demonstrated by plasma cortisol and lactate decrease. Nevertheless, this effect progressively disappeared, allowing a plasma glucose and lactate increase after 6 days of exposure. Fish exposed to GWSF exhibited a liver alanine transaminase (ALT) activity increase after a short exposure while the longest exposure revealed liver damage expressed as an ALT activity decrease. A field caging experiment, carried out in a harbor area (Aveiro Lagoon, Portugal), and a complementary laboratory experiment were designed to assess the influence of the daily tide dynamic on polyaromatic hydrocarbon water distribution and effects on liver EROD and ALT activities, as well as ENA frequency. Eels exposed to low- and high-tide harbor waters, in the laboratory, exhibited a similar degree of genotoxicity, whereas clear differences were observed as EROD induction. In the field experiment, caged eels did not display significant responses enhancing the relevance of natural environmental factors on toxicity mechanisms as well as on the apparent lack of toxicity in harbor waters. PMID- 11386718 TI - Hemoglobin concentration and acetylcholinesterase activity of oligochaetes in relation to lead concentration in spiked sediments from Ignacio Ramirez reservoir. AB - This paper presents toxicity and uptake data of Limnodrilus hoffmeisteri generated by lead in systems using natural sediments from Ignacio Ramirez reservoir. Lead uptake, hemoglobin concentration, and acetylcholinesterase activity were studied in L. hoffmeisteri exposed to spiked sediments. All assays of lead uptake were conducted using whole sediments. Sediment texture was also considered. Acetylcholinesterase activity and hemoglobin concentration decreased after treatment with lead. Acetylcholinesterase activity and hemoglobin concentration tests indicated a response to the concentration of lead. These results suggest the usefulness of a diversity of bioassays to evaluate the toxicity of sediments polluted with heavy metals. PMID- 11386719 TI - Cholinergic and behavioral neurotoxicity of carbaryl and cadmium to larval rainbow trout (Oncorhynchus mykiss). AB - Pesticides and heavy metals are common environmental contaminants that can cause neurotoxicity to aquatic organisms, impairing reproduction and survival. Neurotoxic effects of cadmium and carbaryl exposures were estimated in larval rainbow trout (RBT; Oncorhynchus mykiss) using changes in physiological endpoints and correlations with behavioral responses. Following exposures, RBT were videotaped to assess swimming speed. Brain tissue was used to measure cholinesterase (ChE) activity, muscarinic cholinergic receptor (MChR) number, and MChR affinity. ChE activity decreased with increasing concentrations of carbaryl but not of cadmium. MChR were not affected by exposure to either carbaryl or cadmium. Swimming speed correlated with ChE activity in carbaryl-exposed RBT, but no correlation occurred in cadmium-exposed fish. Thus, carbaryl exposure resulted in neurotoxicity reflected by changes in physiological and behavioral parameters measured, while cadmium exposure did not. Correlations between behavior and physiology provide a useful assessment of neurotoxicity. PMID- 11386720 TI - Acetylcholinesterase inhibition in the threeridge mussel (Amblema plicata) by chlorpyrifos: implications for biomonitoring. AB - The effects of chlorpyrifos, an organophosphorus insecticide, were examined on the activity of the nervous system enzyme acetylcholinesterase (AChE) in the threeridge mussel Amblema plicata in a 24-day laboratory test. Thirty-six mussels in each of seven treatments (18 mussels per duplicate) were exposed to chlorpyrifos (0.1, 0.2, 0.3, 0.6, and 1.2 mg/L), a solvent (acetone), and a solvent-free (well water) control for 12, 24, or 96 h. The activity of AChE was measured in the anterior adductor muscle of eight mussels from each treatment after exposure. To assess potential latent effects, six mussels from each treatment were removed after 24 h of exposure and transferred to untreated water for a 21-day holding period; AChE activity was measured on three mussels from each treatment at 7 and 21 days of the holding period. The activity of AChE in chlorpyrifos-exposed mussels did not differ from controls after 12 or 24 h of exposure (t- test, P>0.05), but was significantly less than controls after 96 h (t- test, P=0.01). AChE activity did not vary among mussels at 24 h of exposure (i.e., Day 0 of holding period) and those at Day 7 and Day 21 of the holding period. Overall changes in AChE activity of mussels during the test were unrelated to individual chlorpyrifos concentrations and exposure times (repeated measure ANOVA; (P=0.06). A power analysis revealed that the sample size must be increased from 2 to 5 replicates (8 to 20 mussels per time interval and test concentration) to increase the probability of detecting significant differences in AChE activity. This calculated increase in sample size has potential implications for future biomonitoring studies with chlorpyrifos and unionid mussels. PMID- 11386722 TI - Chlorophyll a fluorescence and photosynthetic activity as tools for the evaluation of simazine toxicity to Protosiphon botryoides and Anabaena variabilis. AB - On studying the effect of simazine on Protosiphon botryoides and Anabaena variabilis, data revealed that chlorophyll a content and dry weight were decreased with the increase in simazine concentration. High concentration of simazine (0.8 mg L(-1)) reduced gross photosynthesis and carbohydrate content, whereas protein content and respiration rate were increased. Algal cell recovery from simazine toxic effect occurred after 2 and 4 days for Anabaena and Protosiphon, respectively, which may be attributed to the difference in algal genotype of the tested organisms. PMID- 11386724 TI - Arsenate toxicity to Pisum sativum: mineral nutrients, chlorophyll content, and phytase activity. AB - A significant mechanism of arsenate toxicity to Pisum sativum is interference with its mineral nutrient balance. This conclusion is supported by assessments made after exposing P. sativum L. cv. "Phenomen" for 12 days to 12.5, 20.8, and 33.3 mg, and for 32 days to 7.5, 22.1, 36.7, and 73.3 mg of sodium arsenate/kg dry wt soil in the greenhouse. At 20.8 mg of arsenate, mobilization of manganese from the cotyledons was significantly increased and that of zinc decreased. Nitrogen accumulated in the roots. On Day 32, at 22.1 mg of arsenate, magnesium, zinc, and manganese contents of the roots increased, but that of phosphorus of the shoot decreased. The distribution pattern and the ratios between individual elements were severely altered. Relatively more arsenic accumulated from the low than the high soil concentrations. Growth of the shoot was more affected than that of the roots. After a 32-day exposure, chlorophyll content of the leaves increased, but the chlorophyll a/b ratio decreased. On Day 12, at 12.5 mg and 20.8 mg of arsenate, in vivo phytase activity was 64 and 66% that of the controls, respectively. PMID- 11386723 TI - Effect of pendimethalin on growth and photosynthetic activity of Protosiphon botryoides in different nutrient states. AB - The effect of pendimethalin on the green alga Protosiphon botryoides was investigated. Results indicate that specific growth rate, cell number, chlorophyll a level, and dry weight yield significantly decrease with increasing pendimethalin concentrations, while protein and carbohydrate contents increase significantly. On the other hand, photosynthetic activity decreases whereas dark respiration increases with high pendimethalin concentrations. High doses of pendimethalin exhibited no clear trend with 77 K fluorescence (Fv/Fm). Increasing nitrate and phosphate levels led to a decrease in cell number, chlorophyll a, and dry weight as compared with the control at high doses of pendimethalin. The results obtained revealed that N:P < 1 increases the inhibitory effect of high doses of the herbicide. PMID- 11386725 TI - Effects of wood-related sterols on the offspring of the viviparous blenny, Zoarces viviparus L. AB - The parent generation of the viviparous blenny, Zoarces viviparus L., were exposed to phytosterols (a) from oogenesis to parturition and (b) from breeding to parturition. The experiments were performed under laboratory conditions in a test unit supplied with continuous renewal of brackish water. After parturition the offspring were further reared either in clean or in phytosterol-contaminated brackish water. The objective was to study the significance of preexposure of the parent on the effects of phytosterols on the offspring in comparison with effects occurring directly on previously unexposed offspring. The phytosterol concentrations used were (a) 0, 10, 20, and 30 microg/L and (b) 0, 10, and 20 microg/L. Offspring exposed in (a) was further reared in clean water and in (b) offspring from parents exposed to 10 microg/L was either exposed further in the same concentration or left in clean water. Offspring from parents exposed to 20 microg/L was further exposed in the same concentration. Finally one group from unexposed parents was exposed to 10 microg/L after birth. The offspring was studied for 6 months after birth. The results showed that blenny offspring are affected by phytosterols at exposure through the parental generation. The results imply that phyto- sterols affect embryological development of the larvae before hatching as well as the levels of circulating hormones of the parent fish. The larvae contained higher levels of phytosterols as did controls and the bile of exposed female fish contained lower levels of phytosterols implying a link between the higher levels in larvae and lower excretion of the females. The growth of the larvae at 10 microg/L was stimulated regardless of whether the larvae were further exposed, indicating that newborn larvae carried within the female are sensitive to exposure to phytosterols. PMID- 11386726 TI - Effect of endosulfan and parathion on energy reserves and physiological parameters of the terrestrial isopod Porcellio dilatatus. AB - The in vivo effects of parathion and endosulfan on the isopod Porcellio dilatatus were investigated. Feeding parameters (food consumption and assimilation rates), growth, and energy reserves (glycogen, lipid, and protein contents) of pesticide exposed isopods were compared with those of control animals. Isopods were exposed to a wide range of concentrations of parathion or endosulfan (0.1, 1, 10, 25, 50, 100, 250, and 500 microg/g of food) for 21 days. The route of uptake of the pesticides was through the diet (alder leaves). Results revealed that parathion induces a significant depression of glycogen, lipid, and protein contents. However, no significant effect was observed on either feeding parameters or growth. Animals fed endosulfan-contaminated food had smaller amounts of glycogen and lipid than control animals, while protein levels were similar in all treatments. Endosulfan also induced a significant decrease in food consumption and assimilation rates at the highest concentrations tested. Growth rate was also significantly affected by endosulfan. These results suggest that the isopod P. dilatatus is a suitable species for use in toxicity tests and that energy reserves could be used as effect criteria in both laboratory and field studies. PMID- 11386727 TI - Induction of micronuclei in eel (Anguilla anguilla L.) by heavy metals. AB - Micronucleus test was performed in situ on eels (Anguilla anguilla) from river sites with different levels of heavy metal pollution (cadmium and mercury). Cadmium content in eel liver but not micronuclei averages in kidney were associated with cadmium content in sediments. Mercury content in liver was not significantly associated with mercury content in sediments. Both cadmium and mercury induced micronuclei expression in eels when injected, the concentration tested being 1.7 mg metal/kg body weight and the micronuclei induction being 2.64 and 2.35 micronuclei per 1000 cells for cadmium and mercury respectively. It was concluded that these heavy metals are genotoxic for European eel, that eel liver metal content is a sensitive indicator for environmental monitoring of cadmium pollution, and that the micronuclei scores in eels are not a sensitive method to detect heavy metals pollution in freshwater ecosytems. PMID- 11386728 TI - Effects of pulp mill effluents and restricted diet on growth and physiology of rainbow trout (Oncorhynchus mykiss). AB - Juvenile female rainbow trout was exposed for 4.5 months (June to October) to two dilutions of untreated and activated sludge treated whole mill effluent from a pulp mill producing bleached ECF pulp. Two controls were used, on fed ad libitum and a second receiving 0.5% feed of the body weight. All effluent exposed groups were fed ad libitum. Mean weight of the fish was measured monthly. At the end of the experiment a number of physiological and biochemical parameters were analyzed in order to establish the physiological status of the exposed fish in comparison with unexposed fish that obtained ad libitum or restricted amount of feed. The fish exposed to treated effluent grew significantly more than ad libitum control fish until August, whereupon growth retarded in fish exposed to the lower effluent dilution (400 v/v). The growth of fish exposed to untreated effluent did not deviate significantly from the control fed ad libitum. The results from the hematological analysis clearly showed that fish fed restricted amount of feed deviated significantly in most parameters compared with the control fed ad libitum. Fish exposed to treated effluent showed a response pattern similar to that of the control fed restricted amount of feed, whereas the fish exposed to untreated effluent showed a response pattern that did not deviate from that of the ad libitum control. The metabolic parameters suggested that fish exposed to treated effluent had a higher metabolic demand than ad libitum control and that the energy allocation at the end of the experiment was directed to processes other than growth. The responses on hematology were mainly a consequence of the increased energy demand and were not primary effects. The implications of using feed related parameters at field studies are discussed. PMID- 11386729 TI - Biochemical factors contributing to response variation among resistant and sensitive clones of Daphnia magna Ssraus exposed to ethyl parathion. AB - Biochemical factors contributing to response variation in five clones of Daphnia magna exposed to ethyl parathion were studied. Differences in sensitivities to ethyl parathion between sensitive and resistant clones varied between four- and ninefold. Acute toxicity and in vivo acetylcholinesterase (AChE) inhibition responses to ethyl parathion were similar, whereas in vitro AChE sensitivities to paraoxon were higher. Thus indicating that, in vivo, not all ethyl parathion present is converted by Daphnia juveniles to active metabolite, or if so, only a proportion of the paraoxon produced is in direct contact with its target. Comparison of AChE activities obtained during in vitro and in vivo exposures with acute responses revealed clonal concordance only between in vivo AChE sensitivities and acute toxicity. This latter finding indicates that clonal differences in ethyl parathion metabolism rather than differences in AChE sensitivities are the most likely explanations of the observed genetic differences in tolerance. PMID- 11386730 TI - Environmental dissipation of fungicide triphenyltin acetate and its potential as a groundwater contaminant. AB - The effects of various factors on fungicide triphenyltin acetate dissipation in selected soil types were studied; temperature is clearly the most important factor of these. Residues of this fungicide in Fangtzupo clay loam and Lanyang loam incubated at 30 degrees C for 32 days (water content 100, 70, or 40% of field capacity) were 25% of the original amounts. In soil samples incubated at 10, 30, or 40 degrees C with 100% field capacity water content, residues were 80, 26, and 5.5% for Fangtzupo soil, respectively, and 83, 26, and 7.0% for Lanyang soil, respectively. No significant difference in dissipation rate was found between sterilized and unsterilized soil, suggesting that microbial degradation is not an important factor in this process. Rapid degradation rates were observed half-lives of 8.3 to 19.4 days in Fangtzupo clay loam and 8.0 to 16.3 days in Lanyang loam at temperatures ranging from 30 to 40 degrees C (soil water content of 100, 70, and 40% of field capacity). Significant increases in half-life (approximately 150 days) occurred at lower temperatures (10 degrees C) in both soil types. The potential of triphenyltin acetate to contaminate groundwater was tested using the behavior assessment and groundwater pollution-potential models. Results indicate that the leaching rate of the fungicide is very slow (1.0 cm day(-1)) under normal conditions. PMID- 11386731 TI - Effects of the hormone mimetic insecticide tebufenozide on Chironomus riparius larvae in two different exposure setups. AB - The effects of the molting-hormone agonistic insecticide tebufenozide on larvae of the midge Chironomus riparius Meigen were tested in two different exposure setups. After static contamination of first-instar larvae the NOEC, LOEC, and LC50 values were 13.2, 17.4, and 21.14 microg/L, respectively. Semistatic exposure of fourth-instar larvae revealed a lower susceptibility of elder larvae (NOEC 30 microg/L, LOEC 60 microg/L, and LC50 81.94 microg/L). In both cases mortality was not immediate; the effects were postponed and almost exclusively linked to the processes of pupation and emergence. Pupal mortality in the semistatic exposure scheme was twice as high in males as in females during a 100 microg/L treatment. This sex-specific effect probably resulted from the endocrine activity of tebufenozide. Its detection underlines the suitability of C. riparius as a model organism for investigating effects of endocrine-disrupting chemicals in aquatic insects. PMID- 11386732 TI - Toxicity, bioaccumulation, and interactive effects of organotin, cadmium, and chromium on Artemia franciscana. AB - The effects of three organotin compounds-trimethyltin chloride, dimethyltin dichloride, and dibutyltin diacetate-and two heavy metals-cadmium and hexavalent chromium-on Artemia franciscana mortality are investigated in this study. Of all the compounds tested in this work, trimethyltin chloride was, by far, the most toxic. The toxicity order for the five compounds was trimethyltin chloride > potassium dichromate > dimethyltin dichloride > dibutyltin diacetate > cadmium chloride. The big difference in toxicity between dialkyltin and trialkyltin was not accompanied by an equally big difference in bioaccumulation. At a Sn concentration in water of 10 mg/L, the bioconcentration factor was 50 for dimethyltin dichloride and 75 for trimethyltin chloride. At a Sn concentration in water of 100 mg/L, the bioconcentration factor for 6 for dimethyltin dichloride and 9 for trimethyltin chloride. The interactive effect of trimethyltin chloride and cadmium, as well as that of trimethyltin chloride and chromium, was found to be synergistic. Also found to be synergistic was the interactive effect of trimethyltin chloride with cadmium and chromium applied together. PMID- 11386733 TI - Tail moment versus tail length--application of an in vitro version of the comet assay in biomonitoring for genotoxicity in native surface waters using primary hepatocytes and gill cells from zebrafish (Danio rerio). AB - In order to investigate the suitability of an in vitro version of the comet assay with primary hepatocytes and gill cells from zebrafish (Danio rerio), cells were isolated by immersion in trypsin/EDTA solution after whole-body perfusion with phosphate-buffered saline. Within the scope of an 18-month biomonitoring study, primary cells were used to identify the genotoxic potential of native water samples from different sites along the major German rivers, Rhine and Elbe, and to evaluate the sensitivity and practicability of the chosen assay. Depending on the endpoint measured, considerable differences were detected with respect to the number of genotoxic surface water samples: Whereas no differences could be recorded for tail moment and relative DNA contents of head and tail, the number of positively tested native surface water samples significantly increased with tail length as endpoint. PMID- 11386736 TI - Polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans as endocrine disrupters--what we have learned from Yusho disease. AB - Polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans (PCDFs) are persistent environmental pollutants. In some areas wildlife reproduction has been affected by these compounds, which are recognized as endocrine disrupters. In 1968 in northern Kyushu in Japan about 2000 people were poisoned by PCBs and PCDFs (pyrolysis products of PCBs) which contaminated rice oil. Their condition was named "Yusho" disease. A similar poisoning by PCBs in Taiwan was named "Yu-Cheng" disease. The major symptoms of Yusho disease were dermal and ocular lesions, but some of the symptoms, such as irregular menstrual cycles and altered immune responses, were notable with respect to the endocrine disrupting activities of PCBs and related compounds. Several important observations relevant to the mechanisms of Yusho have been made from animal studies. For example, a coplanar PCB congener was shown to cause atrophy of the thymus and PCB administration was thought to alter androgen metabolism. The most tragic aspect of Yusho and Yu-Cheng diseases was the exposure of children to PCBs. In the case of Yu-Cheng, children exposed to PCBs in utero and lactationally were reported to have poor cognitive development. Intellectual impairment was also observed in children born to women who had eaten fish contaminated with PCBs in the United States. From animal studies, alterations in thyroid hormone status, modulation of protein kinase C, and changes in dopamine levels, etc. were proposed as the possible mechanisms for the adverse effects of PCBs on brain development. Whereas coplanar PCB and related congeners, e.g., 2,3,7,8-tetrachlorodibenzo-p-dioxin, induce gene expression via a ligand-dependent transactivating factor, the arylhydrocarbon receptor, alternative pathways for gene expression, e.g., c-Src and cross talk with the MAP kinase pathway, are also reviewed with respect to understanding the toxic mechanisms of these compounds. Finally, the "precautionary principle" is discussed for prevention of the health hazards caused by exposure to endocrine disrupters. PMID- 11386737 TI - The association between daily mortality and ambient air particle pollution in Montreal, Quebec. 1. Nonaccidental mortality. AB - This study was undertaken to determine whether variations in concentrations of particles in the ambient air of Montreal, Quebec, during the period 1984 to 1993, were associated with daily variations in nonaccidental mortality. Fixed-site air pollution monitors in Montreal provided daily mean levels of various measures of particulates and gaseous pollutants. Total sulfates were also measured daily (1986-1993) at a monitoring station 150 km southeast of the city (Sutton, Quebec). We estimated associations for PM(2.5), PM(10), total suspended particles, coefficient of haze (COH), extinction coefficient, and sulfates. We used coefficient of haze, extinction coefficient, and Sutton sulfates to predict fine particles and sulfates for days that were missing. To estimate the associations between nonaccidental mortality and ambient air particles, we regressed the logarithm of daily counts of nonaccidental mortality on the daily mean levels for the above measures of particulates, after accounting for seasonal and subseasonal fluctuations in the mortality time series, non-Poisson dispersion, weather variables, and gaseous pollutants. There were 140,939 residents of Montreal who died during the study period. We found evidence of associations between daily nonaccidental deaths and most measures of particulate air pollution. For example, the mean percentage increase (MPC) for an increase of total suspended particles of 28.57 microg/m(3) (interquartile range, IQ), evaluated at lag 0 days, was 1.86% (95% confidence interval (CI): 0.00-3.76%), and for an increase of coefficient of haze (IQ=18.5 COH units per 327.8 linear m) the MPC was 1.44% (95% CI: 0.75-2.14%). These results are similar to findings from other studies (the mean percentage increase in nonaccidental deaths for a 100 microg/m(3) increase in daily total suspended particles was 6.7%). We also found increases for fine particles and for inhalable particles, but the confidence intervals included unity. All measures of sulfates showed increased daily mortality; e.g., the MPC for sulfates from fine particles (IQ=3.51 microg/m(3)) was 1.86% (95% CI: 0.40-3.35%). We generally found higher excesses in daily mortality for persons 65 years of age and for exposures averaged across lags 0, 1, and 2 days. The slope of the association between daily mortality and ambient air particles in Montreal, which has lower levels of pollution than most major urban centers, is similar to that reported in most other industrialized cities. This study therefore provides further evidence that the association is linear and that any threshold effect, should it exist, would be found at lower levels of air pollution than those found in Montreal. PMID- 11386738 TI - The association between daily mortality and ambient air particle pollution in Montreal, Quebec. 2. Cause-specific mortality. AB - This study was undertaken to determine whether variations in concentrations of particulates in the ambient air of Montreal, Quebec, during the period 1984 to 1993, were associated with daily variations in cause-specific daily mortality. Fixed-site air pollution monitors in Montreal provided daily mean levels of various measures of particles and gaseous pollutants. Total sulfate was also measured daily (1986-1993) at a monitoring station 150 km southeast of the city (Sutton, Quebec). We used coefficient of haze (COH), extinction coefficient, and sulfate from the Sutton station to predict fine particles and sulfate from fine particles for days that were missing. We estimated associations between cause specific mortality and PM(2.5), PM(10), predicted fine particles and fine sulfate particles, total suspended particles, coefficient of haze, extinction coefficient, and total sulfate measured at the Sutton station. We selected a set of underlying causes of death, as recorded on the death certificates, as the endpoint and then regressed the logarithm of daily counts of cause-specific mortality on the daily mean levels for the above measures of particulates, after accounting for seasonal and subseasonal fluctuations in the mortality time series, non-Poisson dispersion, weather variables, and gaseous pollutants. We found positive and statistically significant associations between the daily measures of ambient particle mass and sulfate mass and the deaths from respiratory diseases and diabetes. The mean percentage change in daily mortality (MPC), evaluated at the interquartile range for pollutants averaged over the day of death and the preceding 2 days, for deaths from respiratory diseases was MPC(COH)=6.90% (95% CI: 3.69-10.21%), MPC(Predicted PM2.5)= 9.03% (95% CI: 5.83- 12.33%), and MPC(Sutton sulfate)=4.64% (95% CI: 2.46-6.86%). For diabetes, the corresponding estimates were MPC(COH)=7.50% (95% CI: 1.96-13.34%), MPC(Predicted PM2.5)=7.59% (95% CI: 2.36-13.09%), and MPC(Sutton sulfate)=4.48% (95% CI: 1.08 7.99%). Among individuals older than 65 years at time of death, we found consistent associations across our metrics of particles for neoplasms and coronary artery diseases. Associations with sulfate mass were also found among elderly persons who died of cardiovascular diseases and of lung cancer. These associations were consistent with linear relationships. The associations found for respiratory diseases and for cardiovascular diseases, especially in the elderly, are in line with some of the current hypotheses regarding mechanisms by which ambient particles may increase daily mortality. The positive associations found for cancer and for diabetes may be understood through a general hypothesis proposed by Frank and Tankersley, who suggested that persons in failing health may be at higher risk for external insults through the failure of regulating physiological set points. The association with diabetes may be interpreted in light of recent toxicological findings that inhalation of urban particles in animals increases blood pressure and plasmatic levels of endothelins that enhance vasoconstriction and alter electrophysiology. Further research to confirm these findings and to determine whether they are causal is warranted. PMID- 11386739 TI - Environmental pesticide exposure as a risk factor for Alzheimer's disease: a case control study. AB - The aim of this study was to evaluate the influence of pesticide exposure on the development of Alzheimer's disease (AD), taking into account the potentially confounding factors (genetic, occupational exposure, and sociodemographic). The 1924 study participants (>70 years old) were randomly selected in the Saguenay Lac Saint-Jean region (Quebec, Canada). The AD diagnosis was established in three steps according to recognized criteria. Sixty-eight cases were paired with a nondemented control for age (+/-2 years) and sex. Structured questionnaires addressed to subjects and proxy respondents allowed a description of the sociodemographic characteristics, lifestyle characteristics, and residential, occupational, familial, and medical histories. Assessment of environmental exposure to pesticides was based on residential histories and the agriculture census histories of Statistics Canada (1971-1991) for herbicide and insecticide spraying in the area. Statistical analyses were performed with a logistic regression, adjusting for potential confounding factors. The results failed to show a significant risk of AD with an exposure to herbicides, insecticides, and pesticides. However, future investigations are needed to establish more precisely the identification, measurement, mobility, and bioavailability of neurotoxic pesticide residues in relation to AD. PMID- 11386740 TI - Measurements of manganese with respect to calcium in histological enamel cross sections: toward a new manganese biomarker. AB - Airborne Mn may become an important route of exposure if the use of Mn-containing gasoline additives becomes more widespread. We report on the measurement of manganese and calcium in histological cross sections of shed deciduous tooth enamel of three human subjects. The goal of this research was to measure Mn in tooth enamel for use as a biomarker in assessing manganese exposure in cross sectional and longitudinal studies. The histological locations can be time specific (analogous to examining growth rings in trees). This technique, which may identify critical windows of exposure, can be important for evaluating potential vulnerability of the fetus and neonate to inhaled or ingested Mn. PMID- 11386741 TI - A retrospective study on the relation between renal dysfunction and cadmium concentration in rice in individual hamlets in the Jinzu River basin, Toyama Prefecture, Japan. AB - Using as an index of exposure the Cd concentration in rice and as an index of health effect the urinary abnormality rate (proteinuria, glycosuria, and proteinuria with glycosuria) in the most systematic and extensive health screening examination conducted in 1967 and 1968 in the Jinzu River basin, Japan (a total of 13,183 subjects), we investigated whether a dose-response relationship exists between the two indices. For subjects that lived in the same hamlet since birth, with low Cd concentrations in rice it was confirmed that renal dysfunction does not develop unless the length of residence is prolonged, whereas with high Cd concentrations in rice renal dysfunction develops even when the length of residence is short. For subjects that lived in the same hamlet for over 30 years and that were aged over 50 years, the urinary abnormality rate in individual hamlets showed significant increases with increases in the mean Cd concentration in rice, demonstrating that a dose-response relationship existed. The allowable values of Cd concentration in rice were estimated to be in the range of 0.05-0.20 ppm, representing values lower than the 0.4 ppm provisionally adopted by the Japanese government. PMID- 11386742 TI - Tibia lead levels and methodological uncertainty in 12-year-old children. AB - In vivo bone lead measurements with 109Cd-based K-shell X-ray fluorescence (XRF) have been used to assess long-term lead exposure in adults. Tibia lead levels were measured in 210 children (106 boys, 104 girls) of 11-12(1/2) years of age in a lead smelter town and in a control (nonexposed) town. Tibia lead levels, methodological uncertainties, and models of some of the factors influencing them are presented. 109Cd-based K-shell XRF tibia lead methodological uncertainty in children is comparable to that in adults. PMID- 11386743 TI - Public bus and taxicab drivers' work-time exposure to aromatic volatile organic compounds. AB - Information on the work-time exposure of public bus and taxicab drivers to volatile organic compounds (VOCs) may be a critical factor in exploring the association between occupational exposure and health effects. Accordingly, this study evaluated the work-time VOC exposure of public bus and taxicab drivers by measurement of six selected aromatic VOC concentrations in the personal air of public bus and taxicab drivers during winter and summer. Two groups of five public bus drivers (smokers and nonsmokers) and two groups of five taxicab drivers (smokers and nonsmokers) were recruited for the study. The taxicab drivers were found to be exposed to higher aromatic compound levels than the bus drivers during their daily work time. The personal exposure of the bus and taxicab drivers was influenced by whether or not they smoked plus the season. It was also established that the potential exposure of bus drivers to aromatic VOCs did not exceed that of an unemployed reference group, whereas the potential exposure of taxicab drivers did. Meanwhile, based on comparison of the calculated in-vehicle concentrations with those from a previous study, the VOC levels inside public buses and taxicabs were found to be lower than those inside automobiles. PMID- 11386744 TI - Distribution of methylmercury and inorganic mercury in neonate hamsters dosed with methylmercury during fetal life. AB - The study of the effects of methylmercury (MeHg) contamination has intensified since the MeHg outbreaks in Japan and Iraq. However, most of these studies used high doses of MeHg to obtain its most severe effects. This work identified the MeHg targets in neonate hamsters after administration of two different single oral doses of radiolabeled methylmercury chloride (Me203HgCl) to two groups of pregnant hamsters. The low-dose group received 0.05 microg of 203Hg as Me203HgCl per gram of body weight and the radioactivity was 6.6x10(3) Bq. The high-dose group received 0.55 microg of 203Hg as Me203HgCl per gram of body weight with 2x104 Bq. In the first day of life, the brain was the organ with the highest Me203Hg concentration, followed by the liver, kidneys, and intestines. However, these values of Me203Hg decreased at different rates in all organs until 12 days of life, whereas total 203Hg in the body did not change during the same period. This suggests an age-dependent Me203Hg body distribution instead of an elimination of Me203Hg from the body. PMID- 11386745 TI - Metabolites of organophosphorous insecticides in urine specimens from inhabitants of a residential area. AB - The most frequently used pesticide in U.S. homes, as well as in schools and day care centers, is chlorpyrifos. In 1998, this insecticide was detected in household dust from the former U.S. Forces housing estates in Frankfurt am Main, Germany, resulting from its earlier use up to 1993, i.e., at least 4 years ago. This led to great concern in the new inhabitants. To investigate their internal exposure to the substance, they were offered the opportunity of taking part in biomonitoring examinations. Children playing on the floor were assumed to be especially at risk due to increased exposure to chlorpyrifos via oral or dermal intake. A total of 1146 inhabitants took part in this voluntary investigation. All of them stated that they had never used chlorpyrifos in their homes. Spot urine samples of the study participants were analyzed for six metabolites of organophosphorous insecticides [dimethylphosphate (DMP), diethylphosphate (DEP), dimethylthiophosphate (DMTP), diethylthiophosphate (DETP), dimethyldithiophosphate (DMDTP), and diethyldithiophosphate (DEDTP)] using a very sensitive gas chromatographic method with mass-selective detection and a limit of detection of 1 microg/L. No evidence was found of increased internal exposure due to former chlorpyrifos application in these homes (>4 years ago), either in children or in adults. The median values and 95th percentiles of the urinary metabolite concentrations in 484 adults were (microg/g creatinine): DMP, 15.5 and 102.5; DMTP, 13.5 and 125.8; DMDTP, <1 and 13.1; DEP, 2.1 and 11.6; DETP, <1 and 6.4; DEDTP, both <1. The urinary metabolite concentrations in children <6 years of age were higher; this was caused mainly by lower creatinine concentrations. To conclude, no increase in internal exposure due to former indoor application of chlorpyrifos could be found, and the reference values published for internal organophosphate exposure in adults in Germany were confirmed. However, as shown in other environmental studies, the urinary excretion of organophosphorous metabolites exceeds dietary intake several fold; this has been estimated from the data in various duplicate dietary studies. This observation calls for further investigation. PMID- 11386746 TI - Validation and calibration of mercury intake through self-referred fish consumption in riverine populations in Pantanal Mato-grossense, Brazil. AB - The objective of most epidemiological studies is to classify/rank people according to their relative quantity of consumption. In the specific case of quantifying fish consumption, in order to assess mercury intake rate, the method used must estimate the individual quantity consumed accurately, or from the self referred quantity it must be possible to estimate the real consumption-termed calibration. Therefore, the objective of this study was to validate and calibrate the 24-h recall method for assessing the amount of fish consumed to estimate the mercury intake in riverine populations from Baixada Cuiabana, in Pantanal Mato grossense, Brazil. The weighed dietary method was used as the reference standard, and the validity of the 24-h recall method was assessed using correlation coefficients (r), regression coefficients (beta), and the intraclass correlation coefficient (ICC). The results showed that the ICC, which assesses the agreement level between measures, was a more adequate measure than the correlation coefficient (r). Therefore, the calibration equation was calculated applying a multiple linear regression model using the consumption mean of the second and third days, which presented the highest ICC, stratified by sex and adjusted by age and season variables. PMID- 11386747 TI - Monitoring beef cattle productivity as a measure of environmental health. AB - Domestic livestock have occasionally been used as sentinels of environmental health. Historically, these studies focused on measuring blood or tissue levels of a specific environmental contaminant. For many environmental exposures, however, there is no appropriate biomarker for exposure. The objective of this study was to examine productivity and health information from domestic livestock as one indicator of the potential long-term environmental impact of natural gas developments in a rural area. Intensive biological accounting methods were developed to measure the health and productivity of cow-calf herds surrounding a new sour natural gas processing plant. From the fall of 1991 through calving 1997, cow production records from 7040 recorded bull contacts were examined from seven area cow-calf herds. Detailed information was also collected on other risk factors known to influence beef herd productivity. The median risks for nonpregnancy, abortion, calving late, stillbirth, and calf mortality for local herds did not differ from those of other published reports. There was no significant change in the risk of nonpregnancy, abortion, calving late, stillbirth, or calf mortality. PMID- 11386749 TI - Homology-driven assembly of a sequence-ready mouse BAC contig map spanning regions related to the 46-Mb gene-rich euchromatic segments of human chromosome 19. AB - Draft sequence derived from the 46-Mb gene-rich euchromatic portion of human chromosome 19 (HSA19) was utilized to generate a sequence-ready physical map spanning homologous regions of mouse chromosomes. Sequence similarity searches with the human sequence identified more than 1000 individual orthologous mouse genes from which 382 overgo probes were developed for hybridization. Using human gene order and spacing as a model, these probes were used to isolate and assemble bacterial artificial chromosome (BAC) clone contigs spanning homologous mouse regions. Each contig was verified, extended, and joined to neighboring contigs by restriction enzyme fingerprinting analysis. Approximately 3000 mouse BACs were analyzed and assembled into 44 contigs with a combined length of 41.4 Mb. These BAC contigs, covering 90% of HSA19-related mouse DNA, are distributed throughout 15 homology segments derived from different regions of mouse chromosomes 7, 8, 9, 10, and 17. The alignment of the HSA19 map with the ordered mouse BAC contigs revealed a number of structural differences in several overtly conserved homologous regions and more precisely defined the borders of the known regions of HSA19-syntenic homology. Our results demonstrate that given a human draft sequence, BAC contig maps can be constructed quickly for comparative sequencing without the need for preestablished mouse-specific genetic or physical markers and indicate that similar strategies can be applied with equal success to genomes of other vertebrate species. PMID- 11386750 TI - Five-color-based high-information-content fingerprinting of bacterial artificial chromosome clones using type IIS restriction endonucleases. AB - We have developed a high-information-content fingerprinting (HICF) system for bacterial artificial chromosome (BAC) clones using a Type IIS restriction endonuclease, HgaI, paired with a Type II restriction endonuclease, RsaI. In the method described, unknown five-base overhangs generated with HgaI are partially or fully sequenced by modified fluorescent dideoxy terminators. Using an in-lane size standard labeled with a fifth dye, fragments are characterized by both the size and the sequence of its terminal one to five bases. The enhanced information content associated with this approach significantly increases the accuracy and efficiency of detecting shared fragments among BAC clones. We have compared data obtained from this method to predicted HICF patterns of 10 fully sequenced BACs. We have further applied HICF to 555 BAC clones to assemble contigs spanning 16p11.2 to 16p13.1 of human chromosome 16. PMID- 11386751 TI - Intragenic deletions at Atp7a in mouse models for Menkes disease. AB - Mottled mice have mutations in the copper-transporting ATPase Atp7a. They are proven models for the human disorder Menkes disease (MD), which results from mutations in a homologous gene. Mottled mice can be divided into three classes: class 1, in which affected males die before birth; class 2, in which affected males die in the early postnatal period; and class 3, in which affected males survive to adulthood. In humans, it has been shown that mutations that lead to a complete absence of functional protein cause classical MD, which is characterized by death of boys in early childhood. We hypothesized that the most severely affected mottled alleles would be the most likely to carry mutations equivalent to those causing classical MD and therefore undertook mutational analysis of several class 1 mottled alleles to assess whether these were appropriate models for the disease at the molecular level. Two novel mutations, a deletion of exons 11-14 in mottled spot and an insertion in exon 10 leading to missplicing in mottled candy, were identified. However, these are both "in-frame" mutations, as are the other eight Atp7a mutations reported to date, and therefore no frameshift or nonsense mutations have yet been associated with the mottled phenotype. This contrasts with the mutation spectrum associated with MD, emphasizing the need for caution when mottled mice are used as models for the clinical disorder. PMID- 11386752 TI - Three loci on mouse chromosome 6 influence onset and final incidence of type I diabetes in NOD.C3H congenic strains. AB - The development of insulin-dependent diabetes mellitus in both human and mouse is dependent on the interaction between genetic and environmental factors. The analysis of newly created NOD.C3H congenic strains for spontaneous and cyclophosphamide-induced diabetes has allowed the definition of three controlling genetic loci on mouse chromosome 6. A NOD-derived susceptibility allele at the Idd6 locus strongly influences the onset of diabetes in spontaneous diabetes. A NOD-derived resistance allele at the Idd19 locus affects the final diabetes incidence observed in both models, while a novel locus, provisionally termed Idd20, appears to control Idd19 in an epistatic manner. Decreased diabetes incidence is observed in CY-induced diabetes when Idd20 is homozygous for the C3H allele, while heterozygosity is associated with an increase in diabetes incidence. The Idd20, Idd19, and Idd6 candidate regions fall respectively within genetically defined intervals of 4, 7, and 4.5 cM on mouse chromosome 6. From our YAC contig, Idd6 would appear to localize within a ca. 1.5-Mb region on distal chromosome 6. PMID- 11386753 TI - Calmin, a protein with calponin homology and transmembrane domains expressed in maturing spermatogenic cells. AB - A cDNA named calmin of approximately 3.2 kb was isolated by RNA differential display applied to developing mouse skin. Calmin cDNA encodes 1021 amino acids with two calponin homology (CH) domains in tandem on the N-terminal side and a transmembrane domain on the C-terminal side. The region covering the CH domains showed a high level of homology with beta-spectrin, alpha-actinin, and dystrophin. Among the proteins with the tandem CH domains, calmin is unique in having a transmembrane domain. Three alternative splicing sites were identified at the 3'-side of calmin, giving rise to polymorphic protein products with or without the transmembrane domain. The calmin transcript was detected in adult testis, liver, kidney, and large intestine; the expression in testis was far stronger than that in the other tissues. In situ hybridization and immunostaining revealed that calmin was expressed in maturing spermatogenic cells at later stages. Human calmin cDNA was also isolated, and its exon/intron organization was determined. PMID- 11386754 TI - A new locus for hereditary gingival fibromatosis (GINGF2) maps to 5q13-q22. AB - Gingival fibromatosis (GINGF) is an oral disorder characterized by enlargement of the gingiva. It occurs either as the sole phenotype or combined with other symptoms. Thus far, one GINGF locus has been mapped on chromosome 2, at 2p21, and a second possible locus has been mapped to 2p13. However, the genes responsible for this disorder have not been elucidated. We identified a four-generation Chinese GINGF family in which the disease manifests within 1 year after birth. After exclusion of the two known GINGF loci in this family, we performed a genome wide search to map the chromosome location of the responsible gene. We identified a new locus, GINGF2, on chromosome 5q13-q22 with a maximum two-point lod score of 4.31 at D5S1721 (theta = 0.00). Haplotype analysis placed the critical region in the interval defined by D5S1491 and D5S1453. Within this region, calcium/calmodulin-dependent protein kinase IV (CAMK4) is a strong candidate. PMID- 11386755 TI - A complex duplication created by gene targeting at the imprinted H19 locus results in two classes of methylation and correlated Igf2 expression phenotypes. AB - Imprinting of the mouse H19 and Igf2 genes is dependent on the presence of an intervening imprinting control region (ICR) situated 2 kb upstream of H19 and approximately 70 kb downstream of Igf2. Several recent studies have provided substantial evidence that the unmethylated maternal ICR acts as an insulator that prevents activation of Igf2 by a suite of enhancers downstream of the H19 gene. The methylated paternal ICR and H19 promoter have no activity, allowing sole activation of Igf2 expression. We have produced mice in which a duplication of the H19/Igf2 ICR produces, in each generation, two classes of methylation levels that correlated with two Igf2 imprinting phenotypes. One hypermethylated class also shows activation of the normally silent Igf2 gene, whereas the other hypomethylated class shows only slight activation of Igf2, in agreement with methylation's role in ICR function. This study describes a rare, possibly unique type of mutation that induces two distinct phenotypes in each generation. PMID- 11386756 TI - Influence of intercodon and base frequencies on codon usage in filarial parasites. AB - Base frequency, codon usage, and intercodon identity were analyzed in five filarial parasite species representing five Onchocercidae genera. Wucheria bancrofti, Brugia malayi, Onchocerca volvulus, Acanthocheilonema viteae, and Dirofilaria immitis gene sequences were downloaded from NCBI, and analysis was performed using locally designed computer programs and other freely available applications. A clear sequence bias was observed among the nematode species examined. At the nucleotide level, AT basepairs were present in gene sequences at higher frequencies than GC. In addition, codons ending in A or T were used proportionately more than those with G or C in the third-codon position. In addition, the amino acids used most often corresponded to codons ending in AT basepairs. Intercodon base proportion was biased in that A was found most often at N4, second only to T in certain specific cases. Since all of these sequence biases were observed in a relatively consistent fashion among all of the organisms studied, we conclude that sequence bias is a genetic characteristic, which is associated with multiple filarial genera. PMID- 11386757 TI - Central nervous system, uterus, heart, and leukocyte expression of the LOXL3 gene, encoding a novel lysyl oxidase-like protein. AB - A BLASTN search using the mouse lor-2 cDNA identified three overlapping ESTs (AI752772, AA852888, and R55706) in the GenBank database. These expressed sequence tags were assembled into a contig of 3121 nucleotides with an open reading frame of 2262 bp. The encoded putative polypeptide of 754 amino acids presented all structural characteristics of the lysyl oxidase (LOX) enzyme family, a copper-binding site with four histidyl residues, the lysyl and tyrosyl residues known to be involved in LOX enzyme in the formation of the quinone cofactor and surrounding sequences, and the cytokine receptor-like domain. In addition, four scavenger receptor cysteine-rich (SRCR) domains were found in the N-terminal region of the protein. The gene encoding this new cDNA, which we have referred to as human lysyl oxidase-like 3 (humanLOXL3), has been mapped to chromosome 2p13.3, overlapping at its 3' end the HtrA2 serine protease gene. The structure of the humanLOXL3 gene was deduced from the BAC clone bac91a19 sequence and contained 14 exons. The expression pattern of this new member of the LOX gene family appears to be different from that of the LOX and LOX-like genes, as the central nervous system, neurons, and also leukocytes expressed humanLOXL3. A BLASTN search of the human EST database indicated the presence of ESTs, corresponding to alternative splice variants of LOXL3, that lacked exon 5 and exon 8. The putative resulting protein retained the region encoding the structural and functional elements of the amine oxidase but the second and fourth SRCR domains were truncated and the potential BMP-1 cleavage site was not present. The presence of domains unrelated to the traditional amine oxidase activity is a strong indication that humanLOXL3 might fulfill other functions in addition to intrinsic enzyme activity. PMID- 11386758 TI - Characterization of the murine Lbx2 promoter, identification of the human homologue, and evaluation as a candidate for Alstrom syndrome. AB - The murine Lbx2 gene is a member of the ladybird family of homeobox genes, which is expressed in the developing urogenital system, eye, and brain. Using transgenic mice, we demonstrate that 9 kb of the 5' flanking region of mouse Lbx2 is able to direct expression of a reporter gene in a tissue-specific manner recapitulating the endogenous expression pattern. This regulatory region provides a novel reagent allowing for transgenic expression in the developing urogenital ridge. In addition, we describe the identification of the human homologue, LBX2. Comparison of the human LBX2 and mouse Lbx2 sequences upstream of the coding regions reveals sequence conservation suggesting conserved regulatory regions. Both the human LBX2 and the mouse Lbx2 genes have similar genomic structures and are composed of two exons separated by an intron. We mapped the mouse Lbx2 gene to 35 cM on chromosome 6 and the human LBX2 gene to a homologous region of chromosome 2p13. This is a candidate region for several inherited disorders, including Alstrom syndrome, a disorder that includes ocular, urogenital, and renal abnormalities. Given the expression pattern of Lbx2, the chromosomal location in humans, and the potential function of mammalian ladybird genes, we have begun to analyze patients with ocular disorders and those with Alstrom syndrome for mutations in LBX2. Although polymorphisms were identified, our results indicate that mutations in the coding region of LBX2 do not account for Alstrom syndrome in the six kindreds analyzed. PMID- 11386759 TI - Mutations in Cdh23 cause nonsyndromic hearing loss in waltzer mice. AB - Mutations at the waltzer (v) locus result in deafness and vestibular dysfunction due to degeneration of the neuroepithelium within the inner ear. Here, we use a positional cloning approach to show that waltzer encodes a novel cadherin (Cdh23), which is most closely related to the Drosophila Fat protein. A single nucleotide deletion in the v(J) allele and a single nucleotide insertion in the v allele are predicted to truncate each protein near the N-terminus and produce a functional null allele. In situ hybridization analysis showed that Cdh23 is expressed in the sensory hair cells of the inner ear, where it has been suggested to be a molecule critical for crosslinking of the stereocilia. In addition, Cdh23 is expressed in the urticulo-saccular foramen,the ductus reuniens, and Reissner's membrane, suggesting that Cdh23 may also be involved in maintaining the ionic composition of the endolymph. Finally, mutations in human CDH23 have recently been described for two loci, DFNB12 and USH1D, which cause nonsyndromic deafness, identifying waltzer as a mouse model for human hearing loss. PMID- 11386760 TI - A novel human gene (SARM) at chromosome 17q11 encodes a protein with a SAM motif and structural similarity to Armadillo/beta-catenin that is conserved in mouse, Drosophila, and Caenorhabditis elegans. AB - A novel human gene, SARM, encodes the orthologue of a Drosophila protein (CG7915) and contains a unique combination of the sterile alpha (SAM) and the HEAT/Armadillo motifs. The SARM gene was identified on chromosome 17q11, between markers D17S783 and D17S841 on BAC clone AC002094, which also included a HERV repeat and keratin-18-like, MAC30, TNFAIP1, HSPC017, and vitronectin genes in addition to three unknown genes. The mouse SARM gene was located on a mouse chromosome 11 BAC clone (AC002324). The SARM gene is 1.8 kb centromeric to the vitronectin gene, and the two genes share a promoter region that directs a high level of liver-specific expression of both the SARM and the vitronectin genes. In addition to the liver, the SARM gene was highly expressed in the kidney. A 0.4-kb antisense transcript was coordinately expressed with the SARM gene in the kidney and liver, while in the brain and malignant cell lines, it appeared independent of SARM gene transcription. The SARM gene encodes a protein of 690 amino acids. Based on amino acid sequence homology, we have identified a SAM motif within this derived protein. Structure modeling and protein folding recognition studies confirmed the presence of alpha-alpha right-handed superhelix-like folds consistent with the structure of the Armadillo and HEAT repeats of the beta catenin and importin protein families. Both motifs are known to be involved in protein-protein interactions promoting the formation of diverse protein complexes. We have identified the same conserved SAM/Armadillo motif combination in the mouse, Drosophila, and Caenorhabditis elegans SARM proteins. PMID- 11386761 TI - Analysis of mouse retinal dehydrogenase type 2 promoter and expression. AB - The mouse RALDH2 gene spans >50 kb, has a structure similar to that of human class 1 aldehyde dehydrogenase genes, and localizes to the central region of chromosome 9 by single-strand polymorphism analysis. Expression of mouse RALDH2 was detected in testis, lung, brain, and heart (Northern blot) and in liver and kidney (RNase protection assays). Expression was not detected by RNase protection assay in testis of vitamin A-deficient rats, and all-trans-retinoic acid dosing did not increase expression in vitamin A-deficient rat testis. A 2.3-kb section of the gene 5' to the transcription start site included neither retinoic acid nor retinoid X response elements, but included TATA and CCAAT motifs and AP, AHR, CREB, ER, Ets, and SREBP sites. The promoter initiated transcription of a luciferase reporter in human embryonic kidney cells (EBNA) and mouse Leydig- (TM3) and Sertoli-derived (TM4) cell lines, but neither all-trans-retinoic acid nor 9-cis-retinoic acid affected reporter transcription. These data suggest that relatively weak RALDH2 expression in vitamin A-deficient testis reflects vastly decreased numbers of germ cells, the major site of expression. PMID- 11386764 TI - Imaging of adult brachial plexus traction injuries. AB - Closed, high-energy transfer traction injuries of the adult brachial plexus lead to rupture or avulsion of the spinal nerves. Accurate preoperative diagnosis is crucial for surgical planning and reconstruction. Myelography, computerised tomographic myelography and magnetic resonance imaging are the main radiological methods for preoperative diagnosis of the lesion. This article reviews the current status of imaging of traction injuries of the adult brachial plexus. PMID- 11386765 TI - Acutrak screw fixation versus cast immobilisation for undisplaced scaphoid waist fractures. AB - Fifty-three patients with less than 14 day-old, undisplaced fractures of the waist of the scaphoid were randomized to two groups. Twenty-eight patients were treated by immobilisation in a below elbow plaster cast for 10 weeks while 25 were treated by percutaneous insertion of an Acutrak standard screw. There were no statistically significant differences between the two treatment groups with regard to either the rate of union or the time to union. Patients who underwent surgery had a significantly better range of motion at 16 weeks but there were no significant differences for grip strength. Acute percutaneous internal fixation of undisplaced scaphoid waist fractures using the Acutrak screw allows early mobilisation without adverse effects on fracture healing. PMID- 11386766 TI - The long term recovery curve in adults after median or ulnar nerve repair: a reference interval. AB - This study presents a predicted five-year reference interval for the outcome following repair of the median or ulnar nerve in adults. Forty-four patients were examined with the use of a recently introduced model instrument for documentation after nerve repair that includes "sensory", "motor", and "pain/discomfort" outcomes which together constitute a summarized "total score". Analysis of the "total score" showed that follow-up time and age significantly influence the outcome. There were obvious inferior "motor" results after ulnar nerve injury, but these did not significantly influence the "total score". Significant improvements in the "total score" were seen throughout the follow-up period. PMID- 11386768 TI - Dorso-ulnar osteocutaneous reverse flow flap of the thumb. AB - Three cases of distal thumb reconstruction with a reverse pedicled osteocutaneous flap taken from the dorso-ulnar aspect of the first metacarpal are presented. Even though the indications are rare, this flap is useful for the reconstruction of distal osteocutaneous defects of the thumb where more complex procedures are not feasible or considered as excessive. PMID- 11386767 TI - Extra-articular transverse fractures of the base of the distal phalanx (Seymour's fracture) in children and adults. AB - A series of 25 extra-articular transverse fractures of the base of the distal phalanx (Seymour's fracture) in children, adolescents and adults are presented. Prior to closure of the distal phalangeal epiphysis, the fracture line is usually entirely through the metaphyses, 1 to 2 mm distal to the growth plate. In adults, the fracture line is just distal to the insertion of the extensor tendon. Eighteen of these fractures were treated by closed reduction and splinting and one of these developed an infection and three had mild residual flexion deformities of the finger. Five fractures were treated by K-wire fixation and all healed without complications. Two adult patients presented 4 to 5 weeks after injury with malunion, and of these one underwent refracturing and K-wire fixation with an excellent result. PMID- 11386769 TI - Comparison of mechanical properties of polyvinylidene fluoride and polypropylene monofilament sutures used for flexor tendon repair. AB - We performed an experimental study to evaluate the mechanical properties of polyvinylidene fluoride sutures and to compare their use with that of standard polypropylene sutures for the circumferential, epitendinous suture of a flexor tendon repair. Polyvinylidene fluoride sutures had a smaller suture diameter, a greater knot pull strength and less delayed extension when under creep testing, than polypropylene sutures. Tendons repaired using polyvinylidene fluoride sutures had significantly greater gap and breaking strengths than those repaired using polypropylene sutures. PMID- 11386770 TI - Biomechanical assessment of a new type of flexor tendon repair. AB - A four-strand adaptation of the Kessler repair is described and the results of biomechanical testing in pig tendons are reported. The strength of our repair was compared against standard Kessler repairs using Ethibond or Ti.cron as the core sutures. The average tensile strength for the Ethibond Kessler repair was 33 Newtons and that of the Ti.cron Kessler repair was 31 Newtons. The average tensile strength for the Evans repair was 52 Newtons. This new method of flexor tendon repair is significantly stronger than the modified Kessler repair and is simpler to use than other multi-strand repair techniques. PMID- 11386771 TI - Early breaking strength of repaired flexor tendon treated with 5-fluorouracil. AB - This study investigated the effect of a single intraoperative application of 5 fluorouracil, which may diminish peritendinous adhesion formation, on the tensile strength of repaired digital flexor tendons after 7, 14 and 21 days of healing. Twenty-seven deep flexor tendons from 14 rabbits were exposed to 5-fluorouracil (50 mg/ml) for 5 minutes immediately after repair whereas matched control tendons were exposed to normal saline. Tensile testing at 7, 14 and 21 days revealed no significant differences in the gap or ultimate strengths of the 5-fluorouracil treated and control tendons. PMID- 11386772 TI - Inhibition of tendon cell proliferation and matrix glycosaminoglycan synthesis by non-steroidal anti-inflammatory drugs in vitro. AB - The purpose of this study was to investigate the effects of some commonly used non-steroidal anti-inflammatory drugs (NSAIDs) on human tendon. Explants of human digital flexor and patella tendons were cultured in medium containing pharmacological concentrations of NSAIDs. Cell proliferation was measured by incorporation of 3H-thymidine and glycosaminoglycan synthesis was measured by incorporation of 35S-Sulphate. Diclofenac and aceclofenac had no significant effect either on tendon cell proliferation or glycosaminoglycan synthesis. Indomethacin and naproxen inhibited cell proliferation in patella tendons and inhibited glycosaminoglycan synthesis in both digital flexor and patella tendons. If applicable to the in vivo situation, these NSAIDs should be used with caution in the treatment of pain after tendon injury and surgery. PMID- 11386773 TI - A comparison of metacarpophalangeal joint silastic arthroplasty with or without crossed intrinsic transfer. AB - Forty-three hands in 36 adults undergoing Silastic interposition arthroplasty of the index, middle, ring and little finger metacarpophalangeal joints for rheumatoid arthritis were randomly allocated to undergo replacement with or without crossed intrinsic transfer. The patients were reviewed at a median of 17 (range, 7-50) months after surgery. The demographic characteristics and pre operative clinical measurements of the two groups were indistinguishable. Both groups showed improvement in ulnar drift and an altered arc, but no change in total range of motion at the metacarpophalangeal joints. Grip strength and pulp to pulp pinch were significantly and comparably improved in both groups. There was no difference in pain scores or perceived function between the treatment groups. It is concluded that crossed intrinsic transfer does not significantly affect the outcome of Silastic interposition arthroplasty of the metacarpophalangeal joints in rheumatoid patients. PMID- 11386774 TI - A conservative approach for proximal interphalangeal joint arthroplasty. AB - Access to the proximal interphalangeal joint of the finger for arthroplasty is difficult without detaching its stabilizers or dividing the tendons that cross it, which then require repair and slow rehabilitation. We describe a method that conserves both, so facilitating post-operative rehabilitation.A C-shaped incision is made on the dorsum of the finger. The lateral bands of the extensor expansion are separated from the central slip proximally to the extensor hood. They are then retracted to expose the condyles of the proximal phalanx, which are excised. The PIP joint is then dislocated between the central slip and a lateral band allowing the remainder of the head to be excised. The middle and proximal phalanges are then prepared to accept the prosthesis. The prosthesis is then inserted and the joint is reduced. The lateral bands of the extensor mechanism are sutured back to the central slip before the skin is closed. PMID- 11386775 TI - Arthroscopy: the giant leap forward in wrist surgery? PMID- 11386776 TI - Diagnostic arthroscopy: indications and interpretation of findings. AB - Many authors recommend arthroscopy for diagnostic or therapeutic purposes. Arthroscopy is a sophisticated investigation, which can cause damage requiring wrist fusion. It allows visualization of findings, but these are subtle and difficult, if not impossible to interpret, since the contralateral wrist cannot be used as a standard for comparison. Furthermore, not all intracarpal ligaments are directly and reliably visible through the arthroscope: for example the scaphoid-trapezial ligament which is essential for the stabilization of the distal part of the scaphoid. Therefore, it is questionable whether we can define arthroscopy as a gold standard of diagnosis just because it permits direct vision, as if what cannot be seen does not exist! Traction, mandatory for examination, induces particular stresses that may distort intracarpal motion and so-called dynamic evaluation. Each author tends to present a specific classification, rendering comprehension even more difficult. Accordingly, teaching and learning become a real challenge. Arthroscopy may be important in particular cases, but it remains only one component of a complex clinical assessment. PMID- 11386777 TI - Wrist arthrodesis with excision of the proximal carpal bones using the Ao/ASIF wrist fusion plate and local bone graft. AB - We present our series of 17 patients who underwent wrist arthrodesis with excision of the proximal row carpal bones using the AO wrist fusion plate and local bone graft obtained from the excised proximal carpal row. All patients were evaluated using a questionnaire to assess pain, function, ability to perform an occupation and satisfaction with the procedure. The mean follow-up was 17 months, at which time all the fusions had united. Clinical outcome scores showed that 14 and 15 of the 17 patients achieved good or excellent results with regard to their current condition and clinical improvement, respectively. Four patients required secondary surgery, two for fractures and two for instability of the distal radio ulnar joint unrelated to the wrist fusion. PMID- 11386778 TI - Carpal impaction with the ulnar styloid process: treatment with partial styloid resection. AB - Carpal impaction with the ulnar styloid process (stylocarpal impaction) occurs less frequently than with the ulnar head (ulnocarpal impaction), and more commonly develops in wrists with negative ulnar variance. Physical examination, radiographic evaluation, and wrist arthroscopy are all helpful in excluding alternative causes of ulnar wrist pain. When an ulnocarpal stress test elicits pain, and radiographs suggest that this is due to carpal impaction with the ulnar styloid, partial resection of the styloid process provides successful treatment, so long as the insertion of the triangular fibrocartilage at the base of the styloid is not disrupted. PMID- 11386779 TI - Percutaneous release of trigger digits. AB - Twenty nine patients with 31 trigger digits were treated by percutaneous release under local anaesthesia using an 18 gauge needle. One patient was lost to follow up, and the remainder were examined at a mean follow-up of 14 months. One patient (one thumb) experienced recurrent symptoms, and required an open release. The remaining 27 patients with 29 trigger digits had complete relief of their symptoms. PMID- 11386780 TI - Treatment of de Quervain's disease:role of conservative management. AB - This retrospective study compares two methods used to treat de Quervain's disease: splintage with oral non-steroidal anti-inflammatory drugs (NSAIDs) and steroid injection. Patients were separated into three groups based on symptom severity: group I-minimal, group II-mild, and group III-moderate or severe. Three hundred and nineteen wrists in 300 patients were followed from one to six years. Fifteen of 17 patients with minimal symptoms were relieved with splintage and NSAIDs. However, only seven of 20 in Group II and two of eight in Group III treated similarly were relieved. Of the 249 patients in Group III treated with injections, 76% were completely relieved, 7% were improved, and 4% were not improved. We conclude that classification of patients' with de Quervain's disease based on their pre-treatment symptoms may assist surgeons in selecting the most efficacious treatment and in providing prognostic information to their patients. PMID- 11386781 TI - The use of a moulded metacarpal brace versus neighbour strapping for fractures of the little finger metacarpal neck. AB - Seventy-three patients with fractures of the neck of the little finger metacarpal were randomized to treatment with a moulded metacarpal brace or neighbour strapping. Sixty-five of these attended for follow-up at 3 weeks. Both treatment modalities permitted a functional range of movement, but patients treated with the metacarpal brace had significantly less pain than those treated with neighbour strapping, and this facilitated an early return to work. PMID- 11386782 TI - A single blind, prospective, randomized trial comparing n-butyl 2-cyanoacrylate tissue adhesive (Indermil) and sutures for skin closure in hand surgery. AB - Fifty patients underwent a variety of hand operations and were randomized for wound closure either with tissue adhesive (Indermil) or sutures. The two treatment groups had similar demographic characteristics and similar outcomes at the 2 and 6 week postoperative assessments which were performed by a designated tissue viability nurse blinded to the method of closure. Five minor wound dehiscences occurred: three in the adhesive group and two in the suture group. No infection occurred in either group. In conclusion, the study demonstrates tissue adhesive is as effective as suture in this type of hand surgery. PMID- 11386783 TI - Closed traumatic rupture of the flexor pulleys of a long finger associated with avulsion of the flexor digitorum superficialis. AB - We report a closed rupture of the second, third and fourth annular pulleys associated with avulsion of the flexor digitorum superficialis tendon in the ring finger of a healthy, 48-year-old patient. It was caused by sudden and violent flexion of the finger and led to a serious impairment of the proximal interphalangeal joint motion, despite physiotherapy and dynamic splinting. The patient was treated surgically, 3 months after the injury, with reconstruction of the second (A2) and fourth (A4) annular pulleys and excision of the distal portion of the superficialis tendon. The final functional result was satisfactory. PMID- 11386784 TI - Primary ulnar head prosthesis for the treatment of an irreparable ulnar head fracture dislocation. AB - We report the case of an irreparable fracture - dislocation of the ulnar head with a concomitant fracture of the radius (Galeazzi lesion), treated by implantation of a Herbert Ulnar Head Prosthesis((R)). A stable distal radio-ulnar joint was achieved by careful dissection of a posterior soft tissue flap and accurate reduction of the radius. PMID- 11386785 TI - A comparison of dominant and non dominant hand strengths. PMID- 11386786 TI - Primary vascularized distal radius bone graft for nonunion of the scaphoid. PMID- 11386787 TI - Regarding the article Carpal confusion: is well-intentioned surgery enough? PMID- 11386788 TI - Re. The "poke test" in suspected nerve compression. PMID- 11386793 TI - Epilepsy, depression and antidepressant drugs. AB - Apart from constituting an important management problem, depression coexisting with epilepsy is also an interesting psychiatric phenomenon, with multiple interacting biological, psychological and social factors involved in its causation. New research approaches to the study of epilepsy and depression, including neuroimaging, neurochemical and neuroendocrine techniques, and the arrival of new classes of antidepressants in recent years, suggest it is timely to reconsider this topic. We review current knowledge of the prevalence and causes of interictal depression in epilepsy, focussing mainly on neurobiological factors, and give an overview of recent concepts concerning the management of depression. We also discuss pharmacological treatment of depression in epilepsy, focussing on the association between antidepressants and seizures, and drug interactions. PMID- 11386794 TI - Haemodynamics of arteriovenous malformations of the brain and consequences of resection: a review. AB - The physiological manifestations of arteriovenous fistulae in humans have been studied since the 18th century. However, confusion regarding concepts of cerebral 'steal', 'normal perfusion breakthrough', and 'congestive hyperaemia' continue. Although the advent of more accurate monitoring of pressures and flows within the brain has provided useful information to help understand some of these proposed pathological hypotheses, disagreement still exists. The purpose of this review is to examine the current physiological data in attempt to explain the clinicopathological manifestations of arteriovenous malformations of the brain and the consequences of their removal. PMID- 11386795 TI - Nerve cell tumours of the cerebrum: variable clinical and pathological manifestations. AB - Nerve cell tumours of the cerebrum tend to display a high degree of morphological variability from case to case, and this leads to poor understanding of these tumours. We retrospectively reviewed the clinical and patho-anatomic features of 16 primary nerve cell tumours of the cerebrum (M:9; F:7; average age at onset: 10.2 years). Intraventricular tumours were not included. In 13 patients epileptic seizures were the only symptoms, while three had headache or hemiparesis. Seven tumours were located in the frontal lobe, four in the parietal lobe, two in the temporal lobe and one each in the fronto-parietal lobes, occipital lobe and the midbrain. Tumours were histologically classified into three groups. In the first group, six tumours had the morphological features of classic gangliocytoma or ganglioglioma. In the second group six cerebral and midbrain tumours were composed of small cells, which showed apparent neuronal differentiation including positive immunoreactivity for synaptophysin and the presence of synaptic structures. These tumours usually involved both the cortex and white matter. In the third group, three tumours were composed of small nerve cells and ganglioid cells. All tumours were relatively well circumscribed, and thus eight tumours were totally removed, five subtotally and three partially. Following surgery, three patients, except one, are alive with stable imaging findings for 4 months - 19.3 years (average 11.6 years) after treatment. While small nerve cell tumours are found throughout the cerebrum and its identification broadens the spectrum of neuronal and mixed neuro-glial tumours, most of these tumours are biologically indolent. PMID- 11386796 TI - Pituitary tumours in adolescence: clinical behaviour and neuroimaging features of seven cases. AB - The clinicopathologic features of seven paediatric patients with pituitary adenomas (2 male, 5 female; mean age 14.3 years) were reviewed. There were three non-functioning adenomas, three prolactinomas, and one growth hormone producing adenoma. Five patients presented with visual field deficits, and six patients had endocrine symptoms, which included menstrual irregularities in all female patients, pubertal delay in two females, and growth delay and gigantism in one case each. On neuroimaging studies, five adenomas showed parasellar extension, while the remaining two prolactinomas were intrasellar microadenomas. While two patients with prolactinomas received good results with bromocriptine treatment alone, the remaining five patients underwent either craniotomy or transsphenoidal surgery. Postoperatively, visual disturbances improved markedly in all patients. Two patients also received replacement hormonal therapy. While six patients have been stable for 3.6 years on average, one non-functioning tumour recurred 2 years after the initial transcranial subtotal resection of the tumour. Although there are still many unknowns concerning the biology and optimal treatments for paediatric pituitary adenomas, many of them are assumed to be relatively rapidly growing tumours, while others merely have an earlier tumour genesis than in adults. PMID- 11386797 TI - Surgical treatment of intracranial cavernous angiomas. AB - We present a surgical series of 35 patients (25 males and 10 females) with histopathologically verified intracranial cavernous angiomas. The 35 malformations were located as follows: 21 were in the cerebral hemispheres; 4 in the lateral ventricles, 4 in the brain stem; and 6 in the cerebellum. Seizures and focal neurological deficits were the main clinical features observed in patients with intracranial cavernous angiomas. A number of these vascular malformations were misdiagnosed by computerized tomography. In the last 10 years, magnetic resonance imaging has been the most sensitive method for detecting these lesions. Thirty-five cavernous angiomas were treated surgically; in 33 patients a complete excision, and in 2 patients subtotal excision were obtained. One of the patients died one year after the operation. The overall outcome was good in all of the 34 remaining patients, resulting in improved seizure control or neurological deficit. The rationale for neurologic differential diagnosis and surgical treatment and follow up results are discussed. PMID- 11386798 TI - Magnetic resonance imaging for surgical consideration of acute head injury. AB - The authors studied the extravasation of contrast enhancement on magnetic resonance (MR) imaging within 6 hours after head injury in 10 patients with 12 intracranial haemorrhagic lesions. The decision for surgical intervention was made by neurological examination, computed tomographic scan, and contrast extravasation on MR imaging. Nine of 10 patients showed extravasation of contrast agent and one patient showed equivocal findings of contrast extravasation. All intracranial lesions with contrast extrav asation led to enlargement in size. With the exception of 1 patient who showed equivocal findings of contrast extravasation, 9 patients needed surgical evacuation of the haemorrhagic lesions. The results of the current study imply that extravasation of contrast medium indicates persistence of post-traumatic bleeding. MR imaging with gadolinium enhancement in acutely head injured patients may be used to predict the development of haemorrhagic lesions and could be helpful in decision making for surgical intervention. PMID- 11386799 TI - Stroke in Indonesia: a first large prospective hospital-based study of acute stroke in 28 hospitals in Indonesia. AB - Stroke is an increasing cause of morbidity and mortality in Indonesia. Data on clinical patterns of hospitalized Indonesian stroke patients are still not available. This study is a part of ASNA (ASEAN Neurological Association) Stroke Epidemiological Stu dy aimed to investigate clinical profile of stroke in seven ASEAN countries with the same protocol. From 2065 acute stroke patients admitted to 28 hospitals all over Indonesia, the mean age was 58.8 (Standard Deviation [SD] 13.3) years (range: 18-95 year s). 12.9% were younger than 45 years, and 35.8% were older than 65 years. There were more men than women. Mean admission post-stroke time was 48.5 h (SD 98.8) (range: 1-968 h). Most of them arrived at hospital more than 6 h from stroke onset. The reasons for delayed admission were unawareness of stroke symptoms and long distance transportation. The most frequent stroke symptoms were motor disability. The most common risk factors were hypertension, heart disease, cigarette smoking and diabetes mellitus. Recurrent stroke was found in nearly 20% of patients. Ischemic stroke was the most frequent and the majority of the study subjects were discharged alive and improved. PMID- 11386800 TI - The expression and changes of heat shock protein 70, MDA and haemorheology in rat cortex after diffuse axonal injury with secondary insults. AB - In the present study the role of heat shock protein 70 (HSP70) expression, changes of malonyldialdehyde (MDA) in rat cortex and haemorheology with time after diffuse axonal injury (DAI) only and DAI with secondary insults (SI) were studied. The rat DAI and DAI with SI model were made according to our previous work and animals were divided into a control and another five injury groups with time after injury. Immunohistochemical assay was used to detect the neuronal expression of HSP70 at 0.5h, 3h, 12h, 24h, 72h after DAI or DAI with SI. In the meantime, the high (etah ) and low whole blood viscosity (etaL ), haematocrit (HCT) and RBC aggregation index (AI = etaL/etah ) were also detected and calculated. MDA in the homogenised brain tissue was assayed by thiobarbituric acid (TBA) reaction. The results showed that HSP70 positive neurons were not detected at 30 minutes, but the number of HSP70 positive neurons begin to increase obviously at 3 hours, reach a peak at 24 hours (P< 0.01), and decrease at 72 hours (P= 0.05) after brain injury. The trend of expression of HSP70 was alike for both DAI only or DAI with SI. Meanwhile, MDA, etah, etaL, HCT and AI changes showed the same tendency. Compared with DAI only group, MDA and blood viscosity indexes in DAI with SI were significantly higher at respective time points (P< 0.01). It is concluded that HSP70 expression, MDA and haemorheology indices increased after brain injury and brain injury with SI. Free radicals and haemorheological changes play an important role in the aggravation of brain damage and HSP70 expression upregulation. PMID- 11386802 TI - The microsurgical anatomy of the premamillary artery. AB - The 50 premamillary arteries (PremA), arising from 39 posterior communicating arteries (PCoA), were examined in injected human brains. The PremA, which commonly was single (71.8%) and less frequently double (28.2%), more often arose from the PCoA (97.4% ) than from the posterior cerebralartery (PCA) (2.6%). The PremA ranged between 280 and 780 microm in diameter. It gave off side branches to the hypothalamus (23.1%), optic tract (10.2%), mamillary body (17.9%) and the crus cerebri (35.9%). The anastomoses involving the extracerebral segment of the PremA were present in 35.9% of the cases. They varied in caliber from 50 to 230 microm. The intracerebral segment of the PremA ranged from 280 to 490 microm in diameter. Our study gives a precise anatomic basis for safer operations on the aneurysms of the posterior communicating artery and adjacent vessels. PMID- 11386801 TI - Intraoperative wake-up procedure with propofol and laryngeal mask for optimal excision of brain tumour in eloquent areas. AB - This is the first thesis describing a new technique for awake craniotomy using a laryngeal mask. Awake craniotomy with propofol infusion has become increasingly popular for the optimal excision of brain tumours located in eloquent areas. During awake craniotomy, tracheal intubation is not performed and propofol infusion is limited to within doses which render the patient just sedated. This asleep-awake procedure is occasionally associated with difficulty in controlling brain volume, especially in patients with a significant mass effect of their brain tumours, since sufficient sedation with propofol tends to cause hypercapnea. We report an intraoperative wake-up procedure employing a laryngeal mask, which enables general anaesthesia to be performed at a sufficient dose of propofol and with control of the brain volume under mechanically assisted ventilation. Before the beginning of cortical mapping, propofol infusion is completely terminated, so allowing the patient to wake up within 5-15 min. Following completion of the tumour excision, general anaesthesia is re-induced at a sufficient dose of propofol. The laryngeal mask can be temporarily removed and repositioned with ease, if necessary. In our experience, this technique is applicable for the optimal excision of brain tumours, especially in patients who are very obese or those who have very large lesions. PMID- 11386803 TI - Penetrating injury of the brain by the burr of a high-speed air drill during craniotomy: case report. AB - The most critical complications of craniotomy are caused by the plunging of a skull drill, and there have been no reports of penetrating brain injury caused by the broken burr of a high-speed air drill during craniotomy. Left orbitofrontal craniotomy was performed to clip two aneurysms in a 44 year old man. While cutting the lateral orbital rim with the burr of an Ultra Power surgical drill system, the burr broke off and penetrated the frontal lobe down to the falx cerebri. There were no discernible contusion or haemorrhage in the brain. The accident occurred because a long burr for the angled attachment of a Surgairtome was erroneously attached to the angled attachment of an UltraPower surgical drill. The mismatch between the steel strength and torque of the two burrs designed for different drill systems seemed responsible for breakage of the burr. We learned a valuable lesson from this failure: that all new instruments, including high-speed air drills, must be studied carefully so that staff become thoroughly familiar with their handling. PMID- 11386804 TI - Pineal cavernous angioma presenting with Parkinsonism. AB - Cavernous angioma of the pineal region is rare, as is brain tumour coincident with Parkinsonism. The authors describe the case of a 55 year old woman with a pineal region cavernous angioma, who subsequently developed Parkinsonism after her increased intracranial pressure was relieved. The cause of Parkinsonian syndrome is unclear, but compression of the posterior thalamus and upper mid brain with congestion of the deep venous system may have caused vascular disturbance of the nigro-striate-pallidal system. Her symptoms gradually improved after total excision of the lesion. PMID- 11386805 TI - Creutzfeldt-Jakob syndrome presenting as epilepsia partialis continua. AB - Creutzfeldt-Jakob disease (CJD) is an uncommon rapidly progressive neurological disorder which can have protean clinical presentations. We report an autopsy proven case of CJD presenting initially as epilepsia partialis continua but then developing the typical clinical features of CJD over several weeks. CJD should be considered in the differential diagnosis of new onset epilepsia partialis continua in adults. PMID- 11386806 TI - Vascular Parkinsonism: a case report and review of the literature. AB - Vascular Parkinsonism (VP) is characterised by sudden onset and rapid progression of clinical symptoms, absent or poor response to dopamine substitution therapy, and postural instability with shuffling gait and absence of tremor, making it a clinically distinct entity from idiopathic Parkinson's disease (IPD). Furthermore, it displays certain typical findings in neurological investigations. We report on a patient presenting features of VP associated with an intracerebral lesion not ascribed to VP to date, namely an isolated ischaemic focal lesion located in the left cerebral peduncle between the substantia nigra and nucleus ruber as evidenced by magnetic resonance imaging (MRI). The pathophysiological organic correlate for contralateral extrapyramidal symptoms in this patient may be an interruption of nigro-thalamic projection, interrupting the final subcortical station in the cortic-striato-pallido-nigro-thalamico-cortical loop central to the pathophysiology of parkinsonian syndromes. Non-response t o levodopa therapy could be a consequence of disruption of the cortico-basal ganglia-cortical loop on account of ischaemic destruction of subcortico-cortical axons, the underlying pathology, therefore, not being the result of a loss of nigral dopaminergic neurons or striatal dopamine deficiency pathogonomonic of IPD. To our knowledge, this is the first case of clinically manifest VP to be described with a single lesion in the contralateral cerebral peduncle between the substantia nigra and nucleus ruber, and suggests alternative intracerebral patterns for the distribution of disease-causing lesions in VP, and possibly new pathophysiological explanations for the nature of this disease. PMID- 11386807 TI - Patients with rheumatoid arthritis adapt differently to repetitive painful stimuli compared to healthy controls. AB - The aim of the study was to investigate whether there are changes of the nociceptive system in patients with chronic inflammatory joint pain. A pain model was used which is based on the recording of cortical chemo-somatosensory event related potentials (CSSERP) after nociceptive stimulation of the nasal mucosa with gaseous carbon dioxide (CO(2)). Twenty-five patients with rheumatoid arthritis (RA) were compared to healthy controls matched for age and gender. Responses to both different intensities of painful stimuli and constant intensities of series of 4 stimuli were analysed. When testing increasing CO(2) concentrations ratings and CSSERP amplitudes increased for both patients and controls. However, when repetitive stimulation was performed with an interval of 2s CSSERP amplitudes N1 were significantly greater in RA patients. It is hypothesized that chronic inflammatory joint pain changes nociceptive processing in terms of generalized changes of the nociceptive system which may amplify chronic pain. PMID- 11386808 TI - Schwannoma of the cervical spine presenting with acute haemorrhage. AB - Acute haemorrhagic presentation of spinal schwannoma is a rare event. A case of cervical spinal schwannoma presenting with spontaneous spinal subdural and intramedullary haemorrhage is described. PMID- 11386809 TI - Ganglioglioma of the lateral ventricle presenting with blepharospasm - case report and review of the literature. AB - Gangliogliomas of the lateral ventricle are unusual tumours of the central nervous system. We report a 25-year-old female who presented with a 4-week history of generalized headaches associated with left-sided blepharospasm. Clinical examination revealed bilateral papilloedema but no focal neurological deficits. Computed tomography and magnetic resonance imaging demonstrated a large lesion within the left lateral ventricle. Gross total surgical resection of the lesion was performed through a para-sagittal, trans-callosal approach. Histopathological analysis revealed the diagnosis of ganglioglioma. The post operative course was uneventful, with complete resolution of both the headaches and blepharospasm. The presentation with an ipsilateral blepharospasm has never previously been reported in the setting of a lateral ventricular tumour. PMID- 11386810 TI - Functional magnetic resonance imaging-controlled neuronavigator-guided brain surgery: a case report. AB - The effectiveness of functional magnetic resonance imaging (f-MRI)-controlled and navigator-guided brain surgery for a patient with a recurrent astrocytoma is demonstrated. Preoperative f-MRI was performed in order to identify the motor area and ensure that the tumour was in the left prefrontal area. A more aggressive operation was planned for the recurrent tumour. The f-MRI data were input to the MKM navigation system and during the operation the contours of the tumour and motor area were visualised b y the microscope of the navigation system. The tumour and surrounding gliotic brain tissue were removed completely. The diagnosis was a grade III astrocytoma. The combination of the navigation system and f-MRI was useful for preoperative design of the surgical strategy, and tumour orientation during the operation, enabling aggressive surgery to be performed without functional deficits ensuing. PMID- 11386811 TI - Adult-onset exercise intolerance due to phosphorylase b kinase deficiency. AB - Muscle-specific phosphorylase b kinase deficiency is an unusual form of glycogen storage disorder. The majority of patients are male with an age at diagnosis between 15 to 36 years. Clinical features include exercise intolerance, myalgia and muscle weakness. A forearm ischaemic exercise test is usually normal and histochemical staining for myophosphorylase positive. The demonstration of reduced muscle phosphorylase b kinase activity by biochemical assay confirms the diagnosis. We report a 36 year old male with phosphorylase b kinase deficiency and symptom onset in adult life. PMID- 11386813 TI - Deep brain stimulation: the case for. PMID- 11386812 TI - Prolactinoma with preferential infrasellar extension: a report of two cases. AB - The authors describe two male patients with prolactinomas which were incidentally found either during the course of a complete work up for headache or after minor head trauma. Since these tumours were located mainly in the sphenoid bone and did not show any upward growth to the suprasellar region, they were initially thought to be a primary skull base tumour; however, the serum prolactin levels of these patients were 1,179 ng/ml and 3,260 ng/ml, respectively. The authors thus emphasise the need to recognise this peculiar infrasellar growth pattern of prolactinoma. PMID- 11386814 TI - Deep brain stimulation is superior to ablative surgery for Parkinson's disease: the case against. PMID- 11386816 TI - Grading of meningiomas. PMID- 11386817 TI - Meningiomas invading the sagittal or transverse sinuses, resection with venous reconstruction. AB - Surgery of meningiomas involving dural sinuses leaves the surgeon confronted with a dilemma: leave the fragment invading the sinus and have a higher risk of recurrence, or attempt a total removal with or without venous reconstruction and expose the patient to a potentially greater operative danger. The authors report a series of 80 meningiomas (72 of the sagittal sinus, 5 of the transverse sinus and 3 of the torcular) in whom gross total removal was achieved in all but 7 cases (i.e., 91%), and venous reconstruction attempted in a majority. In total, 70 patients (87.5%) had a good outcome and resumed their previous activities. There was a permanent neurological deficit in seven (8.7%) due to infarction secondary to injury of central veins (all but one in the sagittal sinus mid third). Three patients (3.6%) died from brain swelling; all with meningioma totally occluding the sinus and in whom resection was achieved without sinus reconstruction. There were two recurrences (2.5%) in this series which has a mean follow-up of 8.5 years. The conclusion is to favour, whenever possible, total removal with sinus reconstruction, using a patch for meningiomas with partial sinus invasion and a venous bypass for those with total sinus occlusion. PMID- 11386818 TI - Radiosurgery as alternative treatment for skull base meningiomas. AB - The effect of radiosurgical treatment of skull base meningiomas in 197 patients with a follow-up of at least 2 years was evaluated. Ninety-two of these patients had combined surgical and radiosurgical treatment, while Gamma Knife Radiosurgery (GKRS) was performed as primary treatment in 105 patients. Follow-up was available in 164 patients with intervals of 25-97 months (median 55 months) after GKRS. The imaging controls revealed decreased tumour size in 84 patients (51%), stable tumour volume in 76 ca ses (47%) and increased tumour size in 4 cases (2%). Neurological examinations showed improved neurological status in 58 cases (35%), stable clinical status in 100 patients (61%) and slight worsening in 6 cases (4%). Due to excellent tumour control rate, good clinical outcome and a low complication rate GKRS represents not only an attractive additional treatment option for basal meningiomas, but may even replace microsurgery in selected cases. PMID- 11386819 TI - Surgical treatment of falcotentorial meningioma. AB - Meningiomas arising from the falcotentorial junction are rare. Among our surgical experience of 375 meningiomas, only 4 cases of so-called falcotentorial meningiomas were encountered. We present these four surgical cases. An occipital transtentorial approach was used in three cases, and a combined midline occipital and suboccipital approach in one case. Total tumour excision was impossible in two cases because of engulfing deep venous structures including the great vein of Galen. Postoperative Gamma knife radiosurgery was performed in these two cases. On the other hand, a posteriorly located tumour was relatively easy to remove, and macroscopic total removal was accomplished. In conclusion, precise microvascular anatomical knowledge is indispensable to satisfactorily excise meningiomas in the falcotentorial area without significant morbidity. PMID- 11386820 TI - Brain oedema in secretory meningiomas. AB - Brain oedema is usually associated with intracranial meningiomas in about 50-66%. As underlying causes, different factors like localisation, vascular supply, angiogenic growth factors and histological subtypes are discussed, and its existence is probably multifactorial. We present 11 patients with the rare subtype of secretory meningiomas. Brain oedema was observed in 82%. These tumours are localised mainly at the frontal convexity and at the sphenoid ridge. All 11 patients were female so that hormonal factors also may play a role in the production of peritumoural oedema. The postoperative outcome was good and no recurrences were seen during follow-up. PMID- 11386821 TI - Predictability of internal carotid artery (ICA) dissectability in cases showing ICA involvement in parasellar meningioma. AB - The present study identified predictors for surgical internal carotid artery (ICA) dissection in cases showing ICA involvement in parasellar meningiomas. Twelve cases encountered over the past 4 years were reviewed. Based on MRI findings, patients were divided into two groups; six patients demonstrated complete ICA involvement (encasement) and the other six showed partial ICA involvement (engulfment). The ICA was dissected in all cases in the engulfment group and in four of six cases in the encasement group. The ICA can be dissected even if it is involved at the centre of the tumour if the tumour is soft and can be aspirated. Preservation of the perforating arteries is more important and more difficult. Angiographic finding showing encasement of a long segment of the ICA is unfavourable because of the high possibility that the perforating arteries are involved. Local stenosis of ICA is another unfavourable finding for surgical dissection because tumour invasion of the arterial wall would be suspected. Thus, ICA encasement by the tumour is the less favourable finding for surgery but it is not a decisive predictor. More important findings for ICA dissection in cases showing ICA encasement are involvement of a long segment of the ICA and local ICA stenosis on angiogram. PMID- 11386822 TI - Treatment of tuberculum sellae meningiomas:a long-term follow-up study. AB - Surgical techniques and their results for tuberculum sellae meningiomas were studied. Thirty-three cases, the first of which was operated in 1980, were analysed. There were 4 men and 29 women with an average age of 46.7 years. Eight cases underwent reoperations. The mean follow-up was 10.7 years. Approaches were pterional for 15 patients, FOZ/FO for 10, bilateral subfrontal for 6, and others for 2. Simpson's grade (I, II, III, IV) were 12, 9, 0, and 12 cases respectively. Recurrence rate was 0% for grade I and 58.3% for grade IV. The FOZ/FO approach resulted in a lower Simpson's grade (P=0.05), but other factors were not related to Simpson's grade (P=0.05). The postoperative visual outcome did not depend on total (grade I and II) or subtotal (grade III and IV) removal (P=0.01). We conclude that radical removal of the tumours may result in lower recurrence rate without increasing surgical complications. Furthermore, skull base approaches can improve the rate of radical removal of tuberculum sellae meningiomas. PMID- 11386823 TI - Surgical tactics and outcome of treatment in jugular foramen schwannomas. AB - Seven patients with schwannomas of the jugular foramen were included our study in Samsung Medical Center between 1995 and 1999. Patients with neurofibromatosis were excluded. The records of the seven patients (six surgical case and one nonsurgical case) were retrospectively reviewed. There were six women and one man (mean age, 47 years) with a symptom duration ranging from 3 months to 14 years (mean, 47 months). The predominant symptoms were hearing difficulty, hemifacial spasm and hoarseness. Preoperative audiologic evaluation, computerised tomography (CT), magnetic resonance (MR) imaging, and angiography were performed in most patients. We classified tumours into four types using Kaye and Pellet classification on the basis of radiological and surgical findings. The tumours were: Type A (at cerebellopontine angle) in one; Type B (foraminal) in two; Type C (extracranial and/or foraminal) in two; and Type D (intra- and extracranial) in two cases. We used various surgical approaches such as retrosigmoid suboccipital craniectomy for Type A tumours, infratemporal fossa type A approach (ITFA) for Type C tumours, petro-occipital transsigmoid approach or modified transcochear approach for Type D tumours and ITFA with partial labyrinthectomy for Type B. In the selection of surgical approaches, we took consideration of tumour extension, tumour size, and preoperative hearing function. Facial nerve transposition was not used only in one case of ITFA because of small tumour size (1.5cm). Gross total removal was achieved in five cases, and subtotal removal in one case (Type D tumour) with a single-stage operation. Stereotactic radiosurgery was performed on residual mass in the subtotally removed case. Follow-up period ranged from 13 to 49 months (mean, 27.5 months). There was neither postoperative mortality nor recurrence on follow-up MR imaging. There were two cases of temporary facial nerve palsy and one aggravation of pre-existing low cranial palsy. Two case of sustained vocal cord palsy underwent thyroplasty, but there was no aspiration pneumonia. Persistent cerebrospinal fluid collection was improved with lumboperitoneal shunt. The surgical approaches of each case should be tailored according to their shape and the clinical manifestation. We obtain acceptable outcomes from one-stage operation. PMID- 11386824 TI - Multiple meningiomas of different pathological features: case report. AB - Meningioma is a common intracranial tumour and it may occur not infrequently as one of the multiple tumours, especially in patients with neurofibromatosis. The incidence of multiple meningiomas (MMs) without the stigmata of neurofibromatosis is rare, ranging from 1 to 9% of all meningiomas in the literature. Multiple meningiomas with different pathologic features are even rarer, and most of them are benign histologies. The authors report an extremely rare case of MMs which were presented with malignant and benign histological features simultaneously. The underlying mechanism of MM formation is still unclear, however, subarachnoid spread was thought to be the most likely mechanism. The findings of most of MMs showed identical histopathological features and several molecular biologic studies provided evidence for the monoclonal origin of MMs to back up the above hypothesis. However, different histological features among the reported multiple meningioma cases including our particular one, suggests their origin from multicentric neoplastic foci activated by a supposed tumour-producing factor. However, we cannot completely exclude the possibility of independent progression from monoclonal origin. PMID- 11386825 TI - Expression of the proliferating cell nuclear antigen and clinicopathological features in intracranial meningiomas. AB - PC10, a monoclonal antibody (mAb) to proliferating cell nuclear antigen (PCNA) is known to show immunoreactivity in paraffin-embedded specimens. The authors present the relation between PCNA expression and clinicopathological features in 38 intracranial meningiomas. PCNA scores were obtained by immunohistochemical staining of the paraffin-embedded sections using a streptavidin-biotin immunoperoxidase method with PC10 mAb. Univariate analysis showed that high PC10 scores were associated with old age (> or = 50 years old), male, recurrent tumours, and meningothelial type. However, these high scores did not reach a statistical significance (P> 0.05). PC10 scores of the basal meningioma tended to be higher than that of the hemispheric meningioma (P< 0.05). The staining intensity of PCNA was also markedly increased in basal meningiomas. It is suggested that the proliferative potential is higher in basal meningiomas than in hemispheric meningiomas. Moreover, these results could reflect high recurrence and difficulty in management of the skull base meningiomas. PMID- 11386826 TI - Expression of PDGF and its receptor as well as their relationship to proliferating activity and apoptosis of meningiomas in human meningiomas. AB - In this study, we detected the expression of platelet-derived growth factor (PDGF) and its receptor in 61 human meningiomas by immunohistochemistry and in situ hybridisation. The results showed that almost all the 61 meningiomas expressed PDGFBB and PDGF beta receptor and the positive rate of PDGFAA was 49%. Only two meningiomas expressed PDGF alpha receptor. The positive rate and the immunostaining intensity of PDGFBB and PDGF beta receptor were higher in atypical meningiomas than in benign types. There was no significant difference between the different types of benign meningiomas. The expression of PDGFB chain mRNA was coincident with that of PDGFB chain protein. There was no correlation between the expression of PDGFAA and the types or grades of meningiomas. The correlation between overexpression of PDGFBB/R beta and tumour grade provides a useful parameter in evaluating the prognosis of patients with meningiomas. The proliferative activity of meningiomas was evaluated by the proliferating cell nuclear antigen labelling index (PCNA LI). In the 61 meningiomas, the average PCNA LI (%) was 1.8+/-1.3, 1.9+/-1.3, 1.7+/-0.8 and 11.6+/-5.3 (in fibrous, meningothelial, transitional and atypical meningiomas, respectively). Statistic analysis shows that the PCNA LI is higher in atypical meningiomas than that in benign types, and there was no significant difference between the different types of benign meningiomas. The expression of PDGFBB and PDGF beta receptor was significant enhanced in ascending order from low PCNA LI meningiomas to high ones. This result suggested that PDGFBB/R beta autocrine loop may stimulate the growth of meningiomas. In this study, we also detected the cell apoptosis of meningiomas by terminal deoxynucleotidyl transferase mediated dUTP nick end labelling (TUNEL) method. The average apoptosis labelling index (%) was 0.11+/ 0.05, 0.08+/-0.04, 0.09+/-0.03 and 0.35+/-0.15 in fibrous, meningothelial, transitional and atypical meningiomas respectively. The apoptosis labelling index was higher in atypical meningiomas than that in benign types. There was a positive correlation between the apoptosis labelling index and PC NA LI of meningioma. When the positive rate of PDGFBB and/or PDGF beta receptor was higher in meningioma, the apoptotic cells was also increased. In conclusion, the overexpression of PDGFBB and its relevant receptor PDGFR beta in meningiomas was correlated with grade of meningiomas and the proliferative activity of meningiomas; PDGFBB/R beta autocrine loop may play an critical role in the pathogenesis of meningiomas. PMID- 11386827 TI - A huge frontal meningioma associated with intraoperative massive bleeding and severe brain swelling--case report. AB - A 58 year old female presented with progressive memory disturbance and personality change. Magnetic resonance (MR) imaging disclosed a huge mass lesion accompanied by prominent oedema in the right frontal lobe. Cerebral angiogram demonstrated a vascular-rich tumour and a major drainer through diploic vein. A right frontotemporal craniotomy was performed. We encountered massive bleeding from diploic vein and dura mater immediately at the craniotomy. We were also faced with severe brain swelling at the dural incision. The tumour was solid, highly vascularised, and fairly well demarcated. We performed total removal of the tumour as quickly as possible in order to reduce the intracranial hypertension and avoid the impending brain herniation. The patient had an uneventful recovery and was asymptomatic at 10 months follow-up. PMID- 11386828 TI - Meningioma of the fourth ventricle presenting with intermittent behaviour disorders: a case report and review of the literature. AB - Intraventricular meningiomas are rare, representing 0.5-5% of all intracranial meningiomas. They arise mostly within the lateral ventricles and more rarely in the third ventricle. Meningiomas of the fourth ventricle are exceptional. They are clearly defined as meningiomas arising from the choroid plexus and lying strictly within the fourth ventricle. We report a 76 year old male patient presenting with a 2-week history of headache and cognitive disorders with agitation and restlessness particularly exacerbated at night or when lying down. CT scan and MR imaging showed a contrast-enhancing lesion located purely within the whole fourth ventricle, with slight ventricular enlargement. At surgery, we totally removed a well-vascularised, greyish encapsulated mass attached to the choroid plexus. Pathological examination revealed a WHO grade I fibroblastic meningioma. We reviewed the literature concerning this unusual meningioma location. PMID- 11386829 TI - Epithelioid haemangioendothelioma of the sphenoid bone. AB - The authors report a case of cranial EH occurred in the left sphenoid bone that was totally excised without transfusion. A 26-year-old woman presented with a 1 year history of progressing exophthalmos in the left eye. A cranial X-ray showed a mixed osteolytic and sclerotic expansile mass lesion in the left sphenoid bone. Neuroradiologic imaging study revealed a left temporosphenoidal extra-axial expansile mass lesion with heterogeneous enhancement after contrast enhancement associated with destruction and erosion of the temporal and sphenoid bones. The cerebral parenchyma and orbital contents were compressed without any sign of infiltration. Bone scan showed hot uptake in the left orbital region. Angiogram demonstrated marked neoplastic angiogenesis from the middle meningeal artery and other branches of left external carotid artery, for which preoperative embolisation was tried in vain. Cranio-orbito-zygomatic approach was undergone in two stages because of the patient's religious belief (patient was Jehovah's witness) and profuse bleeding during the first surgery. Two weeks after initial operation, second surgery was carried out with total excision of the residual mass in the left orbital ridge. Histopathological examination revealed typical epithelioid cell cords or nests in myxoid stroma with a positive immunoreactivity to factor VIII-related antigen. The authors report an unusual case of EH involving sphenoid and temporal bone in a young woman. Preoperative embolisation is thought to be absolutely essential before removal. Because there is no convincing data to advise radiation/chemotherapy, total resection and close follow-up may be reasonable. PMID- 11386830 TI - Prevention of postoperative complications in skull base surgery for nasal or paranasal sinus carcinoma invading the skull base. AB - With recent technical advances in skull base surgery, radical resection of a nasal or paranasal sinus carcinoma invading the skull base can now be achieved. To assure a satisfactory surgical result, it is essential to prevent postoperative infection. In our series of 14 cases, serious postoperative infections occurred in the earliest 10 cases, and only 2 of these patients are still alive. The vascularised abdominal muscle flap for skull base reconstruction was fixed with fibrin glue, but was not adequate to fill the dead space, resulting in cerebrospinal fluid leakage and subsequent meningitis. Once the infection occurred, a free bone flap became the focus of infection. Based on these earlier experiences, we used a ROC fastener system to completely fill the dead space with an abdominal muscle flap, and bone flap was primarily craniectomised in the four most recent cases. With this technique, there were no postoperative infections. PMID- 11386831 TI - Surgery on lesions involving cavernous sinus. AB - Selection of the operative route for cavernous sinus related lesions, the extent of the exposure necessary, the need for intraoperative control of the carotid artery and feasibility and need of radical resection depend on the histological nature of the tumour. The age and sex of the patient, principal presenting signs, size of the tumour, extent of cranial nerve and carotid artery involvement, imaging characters and other such features are helpful in estimating the consistency and vascularity of the lesion, site of origin and direction of its spread, and the extent and nature of cavernous sinus involvement. Evaluation of the histology of the lesion on the basis of the radiological and clinical parameters and the impact on decision regarding the surgical strategy is discussed in the present report. PMID- 11386832 TI - Anatomy of the cavernous sinus region. AB - The cavernous region was examined in 20 fetuses, injected with Micropaque, and in 5 adults. The lateral wall of the cavernous region in fetuses was noticed to have four layers. The superficial membrane represents the dural sheath. The second membrane of dense connective tissue involves the trochlear nerve. The third layer, formed by loose connective tissue, involves the oculomotor nerve, and the ophthalmic and maxillary division. The fourth layer, which represents the lateral wall of the cavernous sinus, involves the abducent nerve. The meningohypophyseal trunk can be complete or incomplete. The inferolateral trunk and its branches were found to supply the cavernous portions of the mentioned cranial nerves. The obtained data make the anatomic basis for neurosurgical operations in the cavernous region. PMID- 11386833 TI - Classification of venous ischaemia with MRI. AB - PURPOSE: Venous ischaemia is diagnosed by angiography and estimated with SPECT and PET. But venous ischaemia presents different features due to aetiology, type of onset, time course and collateral circulation. The purpose of this study was to analyse and to classify VI with MRI. METHODS: An analysis of 12 cases of dural arteriovenous fistula (DAVF) with venous ischaemia, 4 cases of sinus thrombosis, and a case of cortical venous thrombosis was performed. Venous ischaemia is classified with MRI as Type 1: no abnormality, Type 2: T2WI showed high signal intensity area and Gd-MRI showed no enhancement, Type 3: T2WI showed high signal intensity area and Gd-MRI showed enhancement, Type 4: venous infarction or haemorrhage. RESULTS: Type 1 was 8 cases. Type 2 was 3 cases and indicated cytotoxic oedema. Type 3 was 2 cases and indicated vasogenic oedema because of the destruction of blood brain barrier. Type 4 was 4 cases. CONCLUSIONS: The classification may be a useful indicator of severity of venous ischaemia and treatment. PMID- 11386834 TI - Skull base techniques for multiple aneurysms in the internal carotid juxta-dural ring region. AB - The aneurysm located in the internal carotid juxta-dural ring region is difficult to surgically obliterate. At surgery, careful drilling of the anterior clinoid process is mandatory, especially when a laterally projecting aneurysm protrudes to or inside the anterior clinoid process.In this paper, treatment procedures using the skull base techniques with intravascular coil embolisation are described by showing a case. PMID- 11386835 TI - The efficacy and safety of transvenous embolisation in the treatment of intracranial dural arteriovenous fistulas. AB - To evaluate the role of transvenous embolisation including its efficacy and safety in the treatment of intracranial dural arteriovenous fistulas (DAVFs), we retrospectively analysed seven cases of intracranial DAVFs treated with transvenous embolisation in combination with arterial embolisation. Four DAVFs were in the cavernous sinus, two in the transverse-sigmoid sinus, and one in the inferior petrosal sinus. The transarterial and transvenous embolic agents included fibred platinum coils (FPC) and interlocking detachable coils (IDC). In all patients, the transarterial embolisation alone had failed to cure the DAVFs. After the combined transvenous embolisation, the anatomical cure was proven in five patients, and all patients were clinically cured. There were no complications in any patient. In conclusion, the transvenous embolisation is a useful and safe approach in the management of intracranial DAVFs. PMID- 11386836 TI - Microcirculatory alterations in a Mongolian gerbil sinus-vein thrombosis model. AB - BACKGROUND AND PURPOSE: The pathophysiology of sinus vein thrombosis (SVT) is still controversial in patients and experimental animals, the microcirculatory alterations in particular. This study was designed to develop a new sinus vein thrombosis model and to further elucidate pathophysiological events such as the relationship between local and regional cerebral blood flow and haemoglobin oxygen saturation (HbSO2), changes of the microvasculature, leukocyte behaviour and brain tissue damage. METHODS: In a first experimental series, animals were divided into two groups which resulted from different procedures of inducing SVT. In the SSS middle occlusion group (SMO group), SVT was induced by the ligation of the superior sagittal sinus right in the middle between the bregma and the confluence sinum. In the SSS posterior occlusion group (SPO group) the ligation was performed close to the confluence sinum. Regional cerebral blood flow (rCBF) was assessed at 36 identical locations by laser Doppler flowmetry together with regional haemoglobin oxygen saturation (HbSO2). In a second series of experiments SVT was induced by ligation of the SSS close to the confluence sinum (SVT group) to study effects on the cortical microcirculation. A sham operation was performed in six animals (sham group). In both groups, an intravital microscopic double tracing technique was utilised for evaluating microvessel structures and leukocyte behaviour. The images were recorded on videotape for evaluating alterations of microvessel (venules, arterioles and capillaries) diameters and numbers of leukocyte rollers and stickers by a digital video analyser. Animals were sacrificed for histological evaluation after 5 days. RESULTS: The posterior sinus ligation caused a significant decrease of rCBF and HbSO2 and brain tissue damage which was not seen in the SMO group. Alteration of rCBF and HbSO2 were positively correlated with infarct size in the SPO group only, where venous infarction was easily reproduced. Therefore, it is suggested that this model is suitable for studying SVT in Mongolian gerbils. Intravital microscopy of the cortical microcirculation revealed no significant changes of vessels diameter in the sham group, whereas a significant dilation of veins and capillaries was seen in the SVT group. Numbers of leukocyte rollers and stickers were positively correlated with infarct size. CONCLUSION: Microcirculatory alterations and brain tissue damage from SVT in the Mongolian gerbil depend on the SSS occlusion site. The newly established mongolian gerbil sinus-vein thrombosis model has advantages compared to previously reported sinus-vein thrombosis models such as easy handling, easy technique, highly reproducibility, and good observation of microcirculatory event. The model allows for studies of cerebral low-flow conditions such as expected to occur in an ischaemic penumbra zone. PMID- 11386837 TI - Surgical strategy for cavernous angioma in hypothalamus. AB - A very rare case of cavernous angioma in the hypothalamus is presented. The patient was successfully treated by total excision of the haematoma and angioma, and there was no regrowth of the lesion on follow-up MRI. Complete excision of the angioma is the recommended surgical strategy even for patients with deep seated lesions. PMID- 11386838 TI - Postoperative cortical venous infarction in tumours firmly adherent to the cortex. AB - It is sometimes difficult to separate extra-axial tumours from the cortical veins in case of tumours attaching tightly to the cortex and the cortical veins. We present two patients having a postoperative cortical venous infarction. A 59 year old female had convexity meningioma above the motor cortex where abnormal cortical anastomotic veins developed. Transient hemiparesis occurred after total removal of the tumour because of venous infarction and cyst formation resulting from sacrifice of these veins which were tightly adherent to the tumour surface. A 15 year old boy with immature teratoma of the pineal region, showing several draining veins around the vascular-rich tumour, presented transient drowsiness, diplopia and partial impairment of bilateral visual acuity postoperatively because of localised cortical brain and venous damage. It is important to make an effort to preserve main cortical veins during operation as much as possible, even if the tumour adheres to the cortical surface. PMID- 11386839 TI - Management of patellofemoral problems. AB - Patellofemoral pain is often a challenge for the physiotherapist, because of its complex aetiology. Physiotherapists with their understanding of soft tissue structure and muscle function are well positioned to effectively manage most patellofemoral problems by improving the extensibility and mobility of tight structures and improving the timing of the elongated muscles. This will involve recognising the biomechanical factors contributing to the symptoms, adequately explaining to the patient the cause of the symptoms and teaching the patient how to manage the symptoms. Specific training of certain muscles with accurate feedback to change the timing of these muscles during functional and sporting activities will be required if this problem is not to recur. PMID- 11386840 TI - Towards a theory of lumbar mobilisation - the relationship between applied manual force and movements of the spine. AB - Optimal use of lumbar mobilisation requires a theoretical knowledge of the mechanical effects that occur. When a mobilisation force is applied the target vertebra will move with accompanying intervertebral displacements at many intervertebral joints. In addition, displacements and deformations of other tissues will contribute to the movement of the skin surface under the therapist's hands. The responses can be quantified in terms of the movements of the skin surface (described by the force-displacement relation), the absolute movement of the target vertebra, or the relative intervertebral movements. There are at least seven variables related to the manner of application of the mobilisation force that can be controlled by the therapist to alter the nature of the response. The patient's response is also determined by a number of variables related to the mechanical properties of the tissues of the spine, extra-spinal structures and the interaction between the patient and the treatment couch. Currently there is a moderate amount of information available about the effects of technique variations but there is very little data to directly link variations in the properties of specific anatomical structures with variations in either absolute or relative spinal movements or tissue loads. PMID- 11386841 TI - Morphology of the cervical intervertebral disc: implications for McKenzie's model of the disc derangement syndrome. AB - Studies which have examined the cervical intervertebral disc have demonstrated that it is not morphologically similar to the lumbar intervertebral disc. Yet review of the clinical literature has revealed that clinicians have often based clinical theories on the assumption that the cervical and lumbar discs do have a similar structure. The purpose of this paper is to review the literature regarding the morphology of the cervical intervertebral disc in relation to McKenzie's clinical theories which claim that the nucleus pulposus of the adult cervical intervertebral disc may be repositioned. Discussion of the proposed biological mechanisms underlying examination and treatment techniques is imperative if the basis of these techniques is to be better understood. PMID- 11386842 TI - A comparison of manual diagnosis with a diagnosis established by a uni-level lumbar spinal block procedure. AB - The study was designed to test if a manipulative therapist (MT) using manual examination techniques alone or when accompanied by a verbal response from the subject as to the pain produced, could diagnose the lumbar segmental level responsible for a subjects low back pain and referred pain. The study consisted of prospective and retrospective parts where the MTs manual diagnosis of the symptomatic lumbar segmental level was compared to a segmental level diagnosis attained by subsequent or previous spinal anaesthetic blocks. In the prospective part of the study the MT's manual diagnosis was 94.12% and 52.9% sensitive in detecting the symptomatic lumbar segmental level with verbal and non-verbal subject responses, respectively. In the retrospective part of the study the MTs manual diagnosis was found to be 60.86% and 47.82% sensitive in detecting the symptomatic lumbar segmental level with verbal and non-verbal subject responses, respectively. In the prospective study the MT's manual diagnosis showed 100% and 80% specificity in detecting subjects with no history of low back pain with verbal and non-verbal subject responses, respectively, and 100% and 75% specificity in the retrospective study with verbal and non-verbal subject responses, respectively. Inter-therapist reliability analysis for the recording of passive physiological intervertebral movements (PPIVMs), passive accessory intervertebral movements (PAIVMs) and 'tissue response' showed percentage agreement rates ranging from 55% to 99%; 74% to 100% and 43% to 100%, respectively, with weighted kappa values ranging from -0.11 to 0.32; -0.15 to 0.24 and kappa values ranging from -0.16 to 0.28, respectively. This study demonstrates that a MT's manual examination when accompanied by a verbal subject response, is highly accurate in detecting the lumbar segmental level responsible for a subjects complaint. PMID- 11386843 TI - Anterior sacrodural attachments - Trolard's ligaments revisited. AB - In contrast to the attention paid to the structures surrounding spinal nerve roots in the intervertebral foramina, the anterior dural attachments are largely ignored, although they have been described since the last decades of the 19th century. These anterior attachments were systematically studied in a series of 30 cadaver dissections and were found to be present in almost 94% of cases. Four types of anterior attachments were observed. The most frequent form (84%) being a system of filaments that present as a double cross vault between the dura mater and the posterior longitudinal ligament extending from L3 to S3 levels. Less frequent were sagittal filaments (30%), short strong ligaments (17%) and a median septum from L3 to the end of the dural sac (7%). No attachments were found in two cadavers. Further studies are needed to clarify the possible role of these structures in transmitting movement to the dural sac and periradicular sleeves when mobilising the last three lumbar vertebrae or the sacrum. PMID- 11386844 TI - Intra-operator and inter-operator reliability of the OSI CA 6000 Spine Motion Analyzer with a new skin fixation system. AB - Following the independent development in the University of Brighton of a new skin fixation system for use with the OSI CA 6000 Spine Motion Analyzer, a series of studies were initiated to test the reliability of the instrument with the new skin fixation system, and the repeatability of the results thus obtained. This paper reports on the outcome of an initial pilot study to compare the existing manufacturer-supplied strap fixation system with the new system. This was followed by a second pilot study to establish single intra-operator reliability utilising 23 asymptomatic subjects. Following successful conclusion of the pilot studies, full inter/intra-operator reliability studies were initiated, utilising 11 asymptomatic subjects. The mean ranges of motion obtained from these results are reported and analyses of variance were used to calculate Intra-class Correlation Coefficients (ICCs) as an indication of reliability. These are summarised as follows: flexion/extension ranges 0.807; lateral flexion ranges 0.923; rotation ranges 0.822. A detailed methodology is explained, along with in depth discussion of the reasoning behind its adoption, and detailed discussion of the results obtained. This study forms a foundation for the establishment of normative data for ranges of motion and patterns of movement in the lumbar spine, as part of a long-term programme of work investigating the effects of low back pain in specific occupational groups on lumbar spine ranges of motion. It is concluded that the results reported justify progression to the collection of normative data for the lumbar spine, utilising the CA 6000 Spine Motion Analyzer with the new skin fixation system, and the widespread application of that data in the clinical setting. PMID- 11386846 TI - Informed consent and manual therapy. PMID- 11386845 TI - Clinical applications of power spectral analysis of electromyographic investigations in muscle function. AB - Electromyographic (EMG) power spectral analysis of the electrical signals produced by a contracting muscle is emerging as a useful clinical tool. It has the potential to measure objectively the fatigue rate of individual muscles during an exertion. This technical note gives a simple introduction to the spectral changes of the EMG signals during fatigue. It outlines the technical aspects of EMG investigations including the data collection, signal analysis and its interpretation. The clinical applications, its present limitations and future applications are also given. The correct use and interpretation of the power spectral analysis in the clinical environment is important for its further development as a clinical measurement. PMID- 11386847 TI - Molecular genetics of type 1 glycogen storage disease. AB - Glycogen storage disease type 1 (GSD 1) comprises a group of autosomal recessive inherited metabolic disorders caused by deficiency of the microsomal multicomponent glucose-6-phosphatase system. Of the two known transmembrane proteins of the system, malfunction of the catalytic subunit (G6Pase) characterizes GSD 1a. GSD 1 non-a is characterized by defective microsomal glucose-6-phosphate or pyrophosphate/phosphate transport due to mutations in G6PT (glucose-6-phosphate translocase gene) encoding a microsomal transporter protein. Mutations in G6Pase and G6PT account for approximately 80 and approximately 20% of GSD 1 cases, respectively. G6Pase and G6PT work in concert to maintain glucose homeostasis in gluconeogenic organs. Whereas G6Pase is exclusively expressed in gluconeogenic cells, G6PT is ubiquitously expressed and its deficiency generally causes a more severe phenotype. Rapid confirmation of clinically suspected diagnosis of GSD 1, reliable carrier testing, and prenatal diagnosis are facilitated by mutation analyses of the chromosome 11-bound G6PT gene as well as the chromosome 17-bound G6Pase gene. PMID- 11386848 TI - Identification of Caenorhabditis elegans isovaleryl-CoA dehydrogenase and structural comparison with other acyl-CoA dehydrogenases. AB - Isovaleryl-CoA dehydrogenase (IVD) is a flavoenzyme, which catalyzes the conversion of isovaleryl-CoA to 3-methylcrotonyl-CoA in the leucine catabolism pathway and transfers electrons to the electron-transferring flavoprotein (ETF). IVDs from human and rat have been identified and characterized previously. In this study, the gene coding for Caenorhabditis elegans IVD has been identified from a published cDNA sequence and molecular modeling has been performed using the human IVD atomic coordinates. The coding sequence for the mature form of the enzyme was expressed in Escherichia coli, and the recombinant nematode IVD enzyme was purified to essential homogeneity. Its spectrum is typical of recombinant FAD containing acyl-CoA dehydrogenases and shows a minor broad absorption band at 650 700 nm characteristic of an IVD:CoA persulfide charge-transfer complex. Following treatment of the enzyme with sodium dithionite to remove the bound CoA persulfide, the K(m) values for isovaleryl-, butyryl-, valeryl-, and hexanoyl-CoA were estimated to be 2.5, 36.2, 10.5, and 33.8 microM, respectively, using the ETF fluorescence reduction assay. The catalytic efficiency (k(cat)/K(m)) for these substrates was 56.9, 1.3, 13.7, and 3.2 microM(-1). min(-1) per mole of FAD, respectively. The apparent binding constant (K(D app)) of the recombinant IVD determined spectrally for isovaleryl-CoA was 0.34 microM. These kinetic parameters confirm that isovaleryl-CoA is the preferred substrate for the purified enzyme. The variability in the protein structure among known and putative IVDs from various species is discussed in the context of possible mechanisms for modulating enzyme activity. PMID- 11386850 TI - Slc19a2: cloning and characterization of the murine thiamin transporter cDNA and genomic sequence, the orthologue of the human TRMA gene. AB - Recently, our group and others cloned the TRMA disease gene, SLC19A2, which encodes a thiamin transporter. Here, we report the cloning and characterization of the full-length cDNA and genomic sequences of mouse Slc19a2. The Slc19a2 cDNA contained a 1494-bp open-reading frame, and had 5'- and 3'-untranslated regions of 189 and 1857 bp, respectively. A putative GC-rich, TATA-less promoter was identified in genomic sequence directly upstream of the identified 5' end. The Slc19a2 gene spanned 16.3 kb and was organized into six exons, a gene structure conserved with the human orthologue. The predicted Slc19a2 protein, like SLC19A2, was predicted to have 12 transmembrane domains and shared a number of other conserved sequence motifs with the human orthologue, including one potential N glycosylation site (N(63)) and several potential phosphorylation sites. Comparison of the Slc19a2 amino acid sequence with those of the other known SLC19A solute carriers highlighted interesting patterns of conservation and divergence in various domains, allowing insight into potential structure-function relationships. The identification of the mouse Slc19a2 cDNA and genomic sequences will facilitate the generation of an animal model of TRMA, permitting future studies of disease pathogenesis. PMID- 11386849 TI - A novel inborn error in the ligand-binding domain of the vitamin D receptor causes hereditary vitamin D-resistant rickets. AB - Mutations in the vitamin D receptor (VDR) cause hereditary vitamin D-resistant rickets (HVDRR), an autosomal recessive disease resulting in target organ resistance to 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)]. In this report, we describe the clinical case and molecular basis of HVDRR in an Asian boy exhibiting the typical clinical features of the disease including alopecia. Using cultured dermal fibroblasts from the patient, 1,25(OH)(2)D(3) resistance was demonstrated by a shift in the dose response required for 25-hydroxyvitamin D-24 hydroxylase (24-hydroxylase) mRNA induction. Western blot showed that the cells express a normal size VDR but contained reduced levels of receptor compared to normal cells. At 24 degrees C, the affinity of the patient's VDR for [(3)H]1,25(OH)(2)D(3) was 50-fold lower than the VDR in normal fibroblasts. Sequence analysis identified a unique T to G missense mutation in exon 6 that changed phenylalanine to cysteine at amino acid 251 (F251C). The recreated F251C mutant VDR showed reduced transactivation activity using a 24-hydroxylase promoter-luciferase reporter. Maximal transactivation activity exhibited by the WT VDR was not achieved by the mutant VDR even when the cells were treated with up to 10(-6) M 1,25(OH)(2)D(3). However, the transactivation activity was partially rescued by addition of RXRalpha. In the yeast two-hybrid system and GST pull-down assays, high concentrations of 1,25(OH)(2)D(3) were needed to promote F251C mutant VDR binding to RXRalpha, indicating defective heterodimerization. In conclusion, a novel mutation was identified in the VDR LBD that reduces VDR abundance and its affinity for 1,25(OH)(2)D(3) and interferes with RXRalpha heterodimerization resulting in the syndrome of HVDRR. PMID- 11386851 TI - Nonradioactive detection of the common Connexin 26 167delT and 35delG mutations and frequencies among Ashkenazi Jews. AB - Mutations in the gap junction beta2 (GJB2) gene, Connexin 26 (Cx26), cause nonsyndromic sensorineural recessive deafness (NSRD). Two frameshift mutations, 167delT and 35delG, are the most frequent Cx26 lesions causing NSRD. The 35delG mutation is panethnic, while the 167delT lesion occurs almost exclusively in the Ashkenazi Jewish population at a carrier frequency of 2 to 4%. To facilitate carrier detection, a simple nonradioactive allele-specific oligonucleotide (ASO) hybridization assay was developed for the 167delT and 35delG mutations. Screening of 1012 anonymous Ashkenazi Jewish individuals from the New York Metropolitan area revealed carrier frequencies for 167delT and 35delG of 3.96% (95% CI: 2.75 5.15%) and 0.69% (95% CI: 0.18-1.20%), respectively. This sensitive, specific, and relatively inexpensive method can reliably identify affected newborns and patients with NSRD as well as facilitate carrier screening for Connexin 26 deafness in the Ashkenazi Jewish community. PMID- 11386852 TI - Is mutated serine hydroxymethyltransferase (SHMT) involved in the etiology of neural tube defects? AB - Neural tube defects (NTD) arise in the first weeks of pregnancy due to a combination of environmental and genetic factors. In mothers of children with NTD elevated homocysteine (Hcy) levels and decreased plasma folate levels were observed, which suggests a defect in the folate-dependent Hcy metabolism. Therefore, mutations in genes coding for enzymes of this metabolism could be involved in NTD. Serine hydroxymethyltransferase (SHMT) catalyzes the reversible reaction of serine and tetrahydrofolate (THF) to glycine and 5,10-methylene THF. Two different isoforms of SHMT are known, one is present in the cytosol (cSHMT) and the other in the mitochondrion (mSHMT). Theoretically, mutated SHMT could lead to elevated Hcy levels and to an altered distribution of the different folate derivatives and might therefore become a risk factor for NTD. This study concerns the molecular genetic analysis of genes coding for both isoforms of the SHMT enzyme by single-stranded conformation polymorphism analysis. Several mutations as well as polymorphisms were found in both genes. The relevance of two variations, the 1420 C>T mutation of the cytosolic isoform and the 4-bp deletion of the mitochondrial isoform (delTCTT 1721-1724), to NTD risk was tested in a study group, which consisted of 109 NTD patients, 120 mothers of children with NTD, and 420 controls. Neither of the two polymorphisms led to an increased risk of NTD. In mothers with the 1420 CC genotype, significant increased Hcy levels are present. Also, significantly decreased red blood cell folate and plasma folate levels were present in individuals with the 1420 CC genotype. Probably, the 1420 C>T polymorphism causes a shift in distribution of the different folate derivatives. The 4-bp deletion of the mSHMT gene did not lead to altered Hcy or folate levels. So far, the results of this study provide no direct evidence for a role of defective SHMT functioning in NTD. Still, the influence of the 1420 C>T polymorphism of the cSHMT gene on the folate-related risk of NTD needs further investigation. PMID- 11386853 TI - Glycerol increases the yield and activity of human phenylalanine hydroxylase mutant enzymes produced in a prokaryotic expression system. AB - Chemical chaperones are low molecular weight compounds known to stabilize proteins in vitro. Recently it was shown that, in transfected cells, these molecules can also correct the defective folding of some mutant proteins. Hyperphenylalaninemia (HPA) has been proposed to be classified as a "conformational disease," since it has been shown that the majority of the PAH mutations affect protein folding, thereby causing an increasing tendency toward aggregation and proteolytic degradation. Based on these observations, the effect of glycerol as a stabilizer agent of recombinant mutant forms of human phenylalanine hydroxylase enzymes (hPAH) produced in a prokaryotic expression system was investigated. The wild-type and two mutant forms of the hPAH protein (R270K and V388M) were expressed in the presence of glycerol in the culture medium. The yield, specific enzymatic activities, and kinetic properties of the recombinant proteins were determined and compared with the data obtained under normal growth conditions. The results obtained demonstrate that glycerol not only improved the yield of the soluble hPAH proteins (2- to 3-fold depending on the mutant enzyme) produced but also increased the specific activity of the purified recombinant enzymes. We speculate that correction of protein folding abnormalities by chemical chaperones may be a possible therapeutic approach to correct conformational diseases. PMID- 11386854 TI - A novel neurodevelopmental syndrome responsive to 5-hydroxytryptophan and carbidopa. AB - Tryptophan hydroxylase (TPH; EC 1.14.16.4) catalyzes the first rate-limiting step of serotonin biosynthesis by converting l-tryptophan to 5-hydroxytryptophan. Serotonin controls multiple vegetative functions and modulates sensory and alpha motor neurons at the spinal level. We report on five boys with floppiness in infancy followed by motor delay, development of a hypotonic-ataxic syndrome, learning disability, and short attention span. Cerebrospinal fluid (CSF) analysis showed a 51 to 65% reduction of the serotonin end-metabolite 5 hydroxyindoleacetic acid (5HIAA) compared to age-matched median values. In one out of five patients a low CSF 5-methyltetrahydrofolate (MTHF) was present probably due to the common C677T heterozygous mutation of the methylenetetrahydrofolate reductase (MTHFR) gene. Baseline 24-h urinary excretion showed diminished 5HIAA values, not changing after a single oral load with l tryptophan (50-70 mg/kg), but normalizing after 5-hydroxytryptophan administration (1 mg/kg). Treatment with 5-hydroxytryptophan (4-6 mg/kg) and carbidopa (0.5-1.0 mg/kg) resulted in clinical amelioration and normalization of 5HIAA levels in CSF and urine. In the patient with additional MTHFR heterozygosity, a heterozygous missense mutation within exon 6 (G529A) of the TPH gene caused an exchange of valine by isoleucine at codon 177 (V177I). This has been interpreted as a rare DNA variant because the pedigree analysis did not provide any genotype-phenotype correlation. In the other four patients the TPH gene analysis was normal. In conclusion, this new neurodevelopmental syndrome responsive to treatment with 5-hydroxytryptophan and carbidopa might result from an overall reduced capacity of serotonin production due to a TPH gene regulatory defect, unknown factors inactivating the TPH enzyme, or selective loss of serotonergic neurons. PMID- 11386855 TI - Methylenetetrahydrofolate reductase polymorphism, type II diabetes mellitus, coronary artery disease, and essential hypertension in the Czech population. AB - Increased plasma concentrations of homocysteine have been found in patients with coronary artery disease (CAD) and essential hypertension (EH) and in patients with diabetic complications. The 677C/T methylenetetrahydrofolate reductase (MTHFR) gene polymorphism is related to the MTHFR enzyme activity and to the plasma homocysteine concentration. This study was designed to investigate an association of this polymorphism with CAD, EH, and type II diabetes mellitus in the Czech population. The MTHFR genotypes were assessed by the polymerase chain reaction-based methodology in a sample of 1199 unrelated Caucasian subjects with CAD, EH, type II diabetes, or a combination of these diseases, and in healthy subjects. Allele frequencies of the MTHFR polymorphism differed considerably between women with and without type II diabetes mellitus (P = 0.00069), with a higher frequency of the C allele in the diabetic women. In addition, the MTHFR T allele frequency was significantly higher in normotensive subjects with CAD compared with normotensive subjects without this disease (P = 0.020). Both associations were confirmed by multiple logistic regressions. In conclusion, while the C allele of the 677C/T MTHFR polymorphism is associated with type II diabetes mellitus in women, the T allele is associated with CAD only in normotensive subjects of Czech origin. PMID- 11386856 TI - Rapid flow-induced responses in endothelial cells. AB - Endothelial cells alter their morphology, growth rate, and metabolism in response to fluid shear stress. To study rapid flow-induced responses in the 3D endothelial cell morphology and calcium distribution, coupled fluorescence microscopy with optical sectioning, digital imaging, and numerical deconvolution techniques have been utilized. Results demonstrate that within the first minutes of flow application nuclear calcium is increasing. In the same time frame whole cell height and nuclear height are reduced by about 1 microm. Whole cell height changes may facilitate reduction of shear stress gradients on the luminal surface, whereas nuclear structural changes may be important for modulating endothelial growth rate and metabolism. To study the role of the cytoskeleton in these responses, endothelial cells have been treated with specific disrupters (acrylamide, cytochalasin D, and colchicine) of each of the cytoskeleton elements (intermediate filaments, microfilaments, and microtubules, respectively). None of these compounds had any effect on the shear-induced calcium response. Cytochalasin D and acrylamide did not affect the shear-induced nuclear morphology changes. Colchicine, however, completely abrogated the response, indicating that microtubules may be implicated in force transmission from the plasma membrane to the nucleus. A pedagogical model based on tensegrity theory principles is presented that is consistent with the results on the 3D endothelial morphology. PMID- 11386857 TI - Utilization of an alternative carbon source for efficient production of human alpha(1)-antitrypsin by genetically engineered rice cell culture. AB - Human alpha(1)-antitrypsin was produced by genetically engineered rice cells using promoter and signal peptide of a rice alpha-amylase isozyme. Batch and continuous cultures were employed to investigate the effects of alternative carbon sources on the alpha(1)-antitrypsin production. While this expression system is inducible by sugar depletion, we have found that the productivity of alpha(1)-antitrypsin increased 2.4- to 3.4-fold, compared with the control medium without carbon source, in medium containing an alternative carbon source, such as pyruvic acid and glyoxylic acid. The accumulated alpha(1)-antitrypsin in the medium containing pyruvic acid reached 18.2-24.2 mg/g-dry cell in 50-70 h by batch culture. PMID- 11386858 TI - Whole-cell immobilization using cell surface-exposed cellulose-binding domain. AB - Specific adhesion of Eshcherichia coli with surface-exposed cellulose-binding domain (CBD) to cellulosic materials was investigated. Whole-cell immobilization was very specific, forming essentially a monolayer of cells onto the different supports. Cells with surface-exposed CBD bound specifically and tightly to cellulose supports at a wide range of pH. In contrast to CBD, which shows the highest binding to cellulose at 4 degrees C, highest cell loading was observed at 37 degrees C. The extent of immobilization was dependent on the amount of surface exposed CBD. Cells binding increased with increasing amount of CBD until binding was saturated. Even induction of very low level of CBD (0.05 mM IPTG) was sufficient to provide specific and tight binding to cellulose support. Because optimal binding can be obtained under physiological conditions such as pH 7 and 37 degrees C, the results demonstrate the general utility of surface-exposed CBD as an efficient means of whole-cell immobilization. PMID- 11386859 TI - Mathematical programming assisted drug design for nonclassical antifolates. AB - A concept from optimization theory, specifically, mathematical programming, is proposed for designing drugs with desired properties. The mathematical programming formulation is solved to obtain the optimal descriptor values, which are employed in the Cerius(2) modeling environment to infer the optimal lead candidates, in the sense that they exhibit both high selectivity and activity while ensuring low toxicity. It has been observed that unique substituent groups and their molecular conformations are responsible for attaining the goal of simultaneous high selectivity and activity. Both linear and nonlinear quantitative structure activity relationships (QSARs) have been developed for use in the proposed approach. A comparative study of these models is done, and it is shown that the QSARs are well represented by nonlinear models. The proposed mathematical programming strategy has been demonstrated for a class of nonclassical antifolates for Pneumocistis carinii and Toxoplasma gondii dihydofolate reductase. Some of the potential leads found in this study have biological properties similar to those in the open literature. We believe the technique proposed is general and can be applied to other structure based drug design. PMID- 11386860 TI - Polyhydroxyalkanoic acids and rhamnolipids are synthesized sequentially in hexadecane fermentation by Pseudomonas aeruginosa ATCC 10145. AB - Rhamnolipids and poly(beta-hydroxyalkanoic acids) (PHAs) are important fermentation products of Pseudomonas aeruginosa. Both contain beta hydroxyalkanoic acids as main constituents. To investigate the possible relationship between their syntheses, we studied the n-hexadecane fermentation by P. aeruginosa (ATCC 10145). PHA synthesis was found to occur only during active cell growth, while substantial rhamnolipid production began at the onset of the stationary phase. The specific synthesis rate of beta-hydroxyalkanoic acids was estimated as 12.6 mg HA/(g dry cells.h) from the PHA formation during the exponential-growth phase. A similar rate was obtained from the beta hydroxyalkanoic acid incorporation in the rhamnolipids produced during the early stationary phase. A regulatory switch of the flow of beta-hydroxyalkanoic acids from PHA polymerization to rhamnolipid synthesis is clearly indicated to occur when the culture reaches the stationary phase. Five rhamnolipid structures were identified using HPLC-MS. Three are monorhamnolipids, two dirhamnolipids. All have a chain of two beta-hydroxyalkanoic acids. The two major components contain only beta-hydroxydecanoic acids; the three minors also have a beta hydroxydecanoic acid linked to the sugar but a beta-hydroxydodecanoic acid or beta-hydroxydodecenoic acid as the second acid. The PHA accumulation reached about 7.5% of the cell dry weight. The monomer composition was relatively constant at different stages of production: in weight fractions, beta hydroxyoctanoic acid, 0.25 (+/-0.05); beta-hydroxydecanoic acid, 0.41 (+/-0.06); beta-hydroxydodecanoic acid, 0.11 (+/-0.05), beta-hydroxytetradecanoic acid, 0.11 (+/-0.06), and beta-hydroxyhexadecanoic acid, 0.12 (+/-0.06). beta Hydroxydecanoic acid was clearly the primary monomer. PMID- 11386861 TI - Permeabilization of metabolites from biologically viable soybeans (Glycine max). AB - Chemical permeabilization has been widely studied for the release useful metabolites from many types of plant cells and tissues. In this study, the effect of 0-30% (v/v) of aqueous methanol solutions were used to permeabilize soybeans for the release of two isoflavonoids: daidzein and genistein. The release of these metabolites increases with increasing methanol concentrations. The amounts of daidzein and genistein released can increase up to 40- and 86-fold, respectively, when incubated in a 30% (v/v) methanol solution for 24 h compared with those incubated with water only. The effect of methanol on the release rates is primarily due to an increase in solubility of the stored daidzein and genistein (14- to 18-fold) inside the seeds, thus maximizing the concentration gradients for metabolite release. However, the viability of the seeds dropped with increase in methanol concentrations and the incubation time. The viability of soybeans (indicated by their ability to germinate) after permeabilization treatment with 0-20% (v/v) methanol solutions was maintained above 80% throughout the 24 h, whereas no seeds were found to be viable when 30% (v/v) methanol solution was used. The permeability coefficients (P) of daidzein and genistein were found to increase as the methanol concentration used was increased. These P values were estimated to range from 1.1 x 10(-)(9) to 1.9 x 10(-)(8) m/s and 1.0 x 10(-)(9) to 1.7 x 10(-)(8) m/s, respectively. The increase in P can be attributed primarily to an increase in the partition coefficient of the metabolites in the soybean seedcoats. An empirical correlation is proposed in which the log P values are described as a function of the metabolite molecular weights and the partition coefficients of the metabolites between octanol and water, K(oct/water), which was modified to include the effect of methanol present. Knowledge obtained from this study will help provide useful selection criteria for chemical permeabilization of plant tissues, such as seeds, with minimal loss in their viability. PMID- 11386862 TI - Scale-up and design of a pilot-plant photobioreactor for the continuous culture of Spirulina platensis. AB - Scale-up of bioreactors has the intrinsic difficulty of establishing a reliable relationship among physical parameters involved in the design of the new bioreactor and the physiology of the cultured cells. This is more critical in those cases where a more complex operation of the bioreactor is needed, such as in photobioreactors. A key issue in the operation of photobioreactors is establishing a quantification for the interaction between external illumination, internal light distribution and cell growth. In this paper an approach to the scale-up of a photobioreactor for the culture of Spirulina platensis, based on a mathematical model describing this interaction, and the operation of a previous reactor 10 times smaller is presented. The paper describes the approach followed in the scale-up, the analysis of different design constraints, the physical realization of the new bioreactor design, innovative use of plastic material walls to improve reactor safety, and finally the corroboration of its satisfactory operation. PMID- 11386863 TI - Characterization of a novel biocatalyst system for sulfide oxidation. AB - It has been demonstrated that an enrichment culture dominated by Thiomicrospira sp. CVO may be cultured on H2S(g) as an energy source under sulfide-limiting conditions in suspended culture with nitrate as the electron acceptor. Hydrogen sulfide (10,000 ppmv) was completely removed from the feed gas and oxidized to sulfate in <3 s of gas-liquid contacting time. Maximum loading of the biomass for sulfide oxidation was observed to be 5.8 mmol H2S/h-g biomass protein, comparable to that reported previously for Thiobacillus denitrificans under similar conditions. However, the enrichment culture was shown to be more tolerant of extremes in pH and elevated temperature than T. denitrificans. Coupled with a reported tolerance of CVO for up to 10% NaCl, these observations suggest that a CVO-based culture is potentially a more robust biocatalyst system for sulfide oxidation than cultures based on Thiobacilli. PMID- 11386864 TI - Properties of RNase A immobilized on magnetic Poly(2-hydroxyethyl methacrylate) microspheres. AB - Magnetic hydrogel microspheres 1.5 microm in size were prepared by dispersion copolymerization of 2-hydroxyethyl methacrylate and ethylene dimethacrylate in the presence of magnetite, which formed the core of the particles. RNase A was coupled to the particles by the cyanuric chloride method. Gel electrophoresis of plasmid DNA pUC 19 (contaminated by bacterial RNA) confirmed RNA degradation with the immobilized enzyme. The effect of temperature and pH on the relative activity of immobilized RNase A was estimated after incubation of the samples at different temperatures (30-80 degrees C) and pH (4.0-8.0). Maximum relative activity was observed at 70 degrees C and pH 6.5. The matrices based on magnetic poly(HEMA) had a low tendency to adsorb RNA. PMID- 11386865 TI - Continuous culture of immobilized streptomyces cells for kasugamycin production. AB - Continuous cultures of immobilized Streptomyces kasugaensis, a kasugamycin producer, were carried out on Celite beads. When using a prototype separator for immobilized-cell separation and recycling, the continuous operation could not be sustained for an extended period as a result of an excessive loss of immobilized cells caused by the poor performance of the separator. Accordingly, the immobilized-cell separator was revised to provide better immobilized-cell settling and thus recycling into the reactor. In a subsequent culture using the revised separator, a stable operation was maintained for over 820 h with a high kasugamycin productivity. The kasugamycin productivity ranged from 9.8 to 16.1 mg/L/h, which was about 14- to 23-fold higher than that in a batch suspended-cell culture. When the original feeding medium concentration was doubled at the end of the continuous culture, the productivity became severely impaired for several reasons, which will be discussed. An excessive formation of free cells and loss of immobilized cells through the separator were also observed. PMID- 11386866 TI - Treatment of high strength distillery wastewater (cherry stillage) by integrated aerobic biological oxidation and ozonation. AB - The performance of integrated aerobic digestion and ozonation for the treatment of high strength distillery wastewater (i.e., cherry stillage) is reported. Experiments were conducted in laboratory batch systems operating in draw and fill mode. For the biological step, activated sludge from a municipal wastewater treatment facility was used as inoculum, showing a high degree of activity to distillery wastewater. Thus, BOD and COD overall conversions of 95% and 82% were achieved, respectively. However, polyphenol content and absorbance at 254 nm (A(254)) could not be reduced more than 35% and 15%, respectively, by means of single biological oxidation. By considering COD as substrate, the aerobic digestion process followed a Contois' model kinetics, from which the maximum specific growth rate of microorganisms (mu(max)) and the inhibition factor, beta, were then evaluated at different conditions of temperature and pH. In the combined process, the effect of a post-ozonation stage was studied. The main goals achieved by the ozonation step were the removal of polyphenols and A(254). Therefore, ozonation was shown to be an appropriate technology to aid aerobic biological oxidation in the treatment of cherry stillage. PMID- 11386867 TI - A multiphasic hollow fiber reactor for the whole-cell bioconversion of 2-methyl 1,3-propanediol to (r)-beta-hydroxyisobutyric acid. AB - This paper describes the bioconversion of 2-methyl-1,3-propanediol to (R)-beta hydoxyisobutyric acid (HIBA) by Acetobacter ALEI in a hollow fiber membrane bioreaction system arrangement that allows the integration of three liquid phases: the aqueous bioconversion phase, the organic phase consisting of a solution of trioctyl phosphine oxide (TOPO) in isooctane, and the third phase consisting of a basic stripping solution that allows reextraction of HIBA from the organic phase. A comparison of HIBA mass transfer experiments was carried out in the membrane reactor with two and three phases for different pH and TOPO concentrations. The use of the three-phase arrangement allows the extraction of high quantities of HIBA from the aqueous medium (higher than 85%) independently of the pH, whereas in the two-phase system the percentage of HIBA extracted from the aqueous medium was lower, 42% in the best case, and strongly influenced by the pH. The percentage of the extractive agent TOPO in the organic phase influenced on the mass transfer rate in both bi- and triphasic arrangements. By simply integrating the re-extraction phase in the system it was possible to increase the extraction yield by 2-fold, reduce the amount of TOPO by 4-fold, and operate at the more favorable pH 4. A bioconversion experiment was done in these conditions (pH = 4, TOPO = 5%) to confirm the advantages of including the third stripping solution. Fed-batch operation of the triphasic membrane reactor was maintained for more than 20 h, reaching an HIBA concentration in the stripping solution of 29 g L(-)(1). PMID- 11386868 TI - Characterization of dicarboxylic acids for cellulose hydrolysis. AB - In this paper, we show that dilute maleic acid, a dicarboxylic acid, hydrolyzes cellobiose, the repeat unit of cellulose, and the microcrystalline cellulose Avicel as effectively as dilute sulfuric acid but with minimal glucose degradation. Maleic acid, superior to other carboxylic acids reported in this paper, gives higher yields of glucose that is more easily fermented as a result of lower concentrations of degradation products. These results are especially significant because maleic acid, in the form of maleic anhydride, is widely available and produced in large quantities annually. PMID- 11386869 TI - Intrinsic oxygen use kinetics of transformed plant root culture. AB - Root meristem oxygen uptake, root tip extension rate, and specific growth rate are assessed as a function of dissolved oxygen level for three transformed root cultures. The influence of hydrodynamic boundary layer was considered for all measurements to permit correlation of oxygen-dependent kinetics with the concentration of oxygen at the surface of the root meristem. Oxygen uptake rate is shown to be saturated at ambient conditions, and a saturation level of approximately 300 micromole O2/(cm(3) tissue.hr) was observed for all three of these morphologically diverse root types. In nearly all cases, the observation of a minimum oxygen pressure, below which respiration, extension, or root growth would not occur, could be accounted for as a boundary layer mass transfer resistance. The critical oxygen pressure below which respiration declines is below saturated ambient oxygen conditions. In contrast, critical oxygen pressures for root tip extension were much higher; extension was nearly linear for the two thicker root types (Hyoscyamus muticus, henbain; Solanum tuberosum, potato) above ambient oxygen levels. The performance of the thinnest root, Brassica juncea (Indian mustard) was consistent with reduced internal limitations for oxygen transport. Extension rates did not correlate with biomass accumulation. The fastest growing henbain culture micro = 0.44 day(-)(1)) displayed the slowest extension rate (0.16 mm/hr), and the slowest growing mustard culture (micro = 0.22 day(-)(1)) had the fastest tip extension rate (0.3 mm/hr). This apparent paradox is explained in terms of root branching patterns, where the root branching ratio is shown to be dependent upon the oxygen-limited mersitem extension rate. The implications of these observations on the performance of root culture in bioreactors is discussed. PMID- 11386870 TI - L-asparaginase release from Escherichia coli cells with K2HPO4 and Triton X100. AB - A method to release L-asparaginase (EC 3.5.1.1) from ATCC Escherichia coli 11303 cells by chemical permeabilization was studied. It was found that a combination of K2HPO4 and Triton X100 was effective. The influences of K2HPO4 concentration, Triton concentration, E. coli cell concentration and pH on the release of enzyme and proteins were investigated in detail. Experimental results showed that 12.5% (w/v) K2HPO4, 2% (w/v) Triton X100 and 3 x 10(8) cells/mL made the amount of enzyme released over 70%. L-Asparaginase in K2HPO4 and Triton solution could remain stable at least for 24 h. The release effect of K2HPO4 and Triton X100 used simultaneously was better than that of K2HPO4 and Triton X100 used separately in succession. Electron microscopy indicated that the chemical treatment altered the surface structure of E. coli cells but did not break them. As the method does not produce a large amount of cell fragments and the amount of enzyme released is relatively high, it can be thought to be an valuable and economic method to release intracellular enzyme. PMID- 11386871 TI - Human chymotrypsinogen B production with Pichia pastoris by integrated development of fermentation and downstream processing. Part 1. Fermentation. AB - Based on an integrated approach of genetic engineering, fermentation process development, and downstream processing, a fermentative chymotrypsinogen B production process using recombinant Pichia pastoris is presented. Making use of the P. pastoris AOX1-promotor, the demand for methanol as the single carbon source as well as an inducer of protein secretion enforced the use of an optimized feeding strategy by help of on-line analysis and an advanced controller algorithm. By using an experimental system of six parallel sparged column bioreactors, proteolytic product degradation could be minimized while also optimizing starting conditions for the following downstream processing. This optimization of process conditions resulted in the production of authentic chymotrypsinogen at a final concentration level of 480 mg.L(-)(1) in the whole broth and a biomass concentration of 150 g.L(-)(1) cell dry weight, thus comprising a space-time yield of 5.2 mg.L(-)(1).h(-)(1). Alternatively to the high cell density fermentation approach, a continuous fermentation process was developed to study the effects of reduced cell density toward oxygen demand, cooling energy, and biomass separation. This development led to a process with a highly increased space-time yield of 25 mg.L(-)(1).h(-)(1) while reducing the cell dry weight concentration from 150 g.L(-)(1) in fed-batch to 65 g.L(-)(1) in continuous cultivation. PMID- 11386872 TI - Human chymotrypsinogen B production from Pichia pastoris by integrated development of fermentation and downstream processing. Part 2. Protein recovery. AB - The purification of human chymotrypsinogen B (hCTRB) after expression and secretion by the yeast Pichia pastoris is described based on two different approaches using integrated initial recovery. Extraction employing aqueous two phase systems (ATPS) from poly(ethylene glycol) and sodium sulfate allows direct processing of cell containing yeast suspensions of 50% wet weight. The target protein is obtained partially purified in the top phase while cells and cell debris are partitioned to the bottom phase of the system. hCTRB is further purified by adsorption from the top phase to the cation exchanger SP Sepharose Big Beads and elution in a salt step. The single step isolation of hCTRB is possible by expanded bed adsorption (EBA) using a fluidized cation exchanger (Streamline SP XL). A design strategy is shown taking both target protein binding and stable fluidization of the stationary phase in cell containing suspensions into consideration. For the example of hCTRB isolation from cell containing P. pastoris suspensions, a successful use of this strategy is demonstrated. Both initial recovery strategies deliver a product that can be further purified and formulated by ultrafiltration/diafiltration followed by lyophilization, resulting in a homogeneous product. Scale-up to 30-90 L of culture suspension was shown for both methods, resulting in a product of similar quality. Comparing both strategies reveals that the two-step ATPS route is better suited for high cell density cultures, while the single step EBA method is preferred for cultures of moderate cell density. This is due to the fact that application of EBA is restricted to suspensions of 10-12.5% wet weight cell concentration, thus necessitating dilution of the original broth prior to sample application. The data presented show that integrated recovery operations are a valuable alternative to traditional processing for systems that are problematic during initial solid-liquid separation. PMID- 11386873 TI - Carbon dioxide induced soybean protein precipitation: protein fractionation, particle aggregation, and continuous operation. AB - A novel protein fractionation technique using a volatile electrolyte has been developed. Carbon dioxide was used to isoelectrically precipitate 80% and 95% pure glycinin and beta-conglycinin fractions from soybean isolate. The protein fractions precipitated as primary particles 0.2-0.3 microm in diameter, which under optimum conditions may be recovered as aggregates up to 500 microm in diameter. The dependency of protein fractionation efficiency on aggregate settling rates has been demonstrated. The isoelectric points of the two main soybean fractions, glycinin and beta-conglycinin, were calculated to be pH 5.2 and 4.95, respectively. Solution pH was accurately controlled by pressure in the isoelectric pH range of the different soybean protein fractions, and a pH "overshoot" was eliminated. Volatile electrolyte technology was also applied to a continuous process in order to eliminate the particle recovery concerns associated with batch precipitation and to demonstrate the potential for scale up. Glycinin was effectively recovered on-line (94% glycinin recovery) with a purity approaching that of the batch process (95%). PMID- 11386874 TI - Effect of adsorbent porosity on performance of expanded bed chromatography of proteins. AB - Expanded bed or fluidized bed adsorption has emerged as an important unit operation in downstream processing of proteins. A number of specifically designed commercial adsorbents are available today for expanded bed purification of proteins. Protein purification essentially requires adsorbent matrices that have large pore size. Very large pore size or macroporous adsorbents can provide high efficiency in packed beds even at high flow rates on account of reduced pore diffusion resistance resulting from finite intraparticle flow in the macropores. This is reflected in leveling off of HETP (height equivalent to theoretical plate) versus flow curve after a threshold velocity. Expanded bed operation, on the other hand, can also show plateauing of the HETP curve, but not necessarily on account of macroporosity of adsorbent. It is shown in this article how any adsorbent intended for protein adsorption in expanded bed mode can give plateauing HETP curve, regardless of pore size. As a result, RTD measurements on an expanded bed can give equal, and at times better, performance than a corresponding packed bed. Large pore size, on the other hand, can result in lesser retention of biomass and easy flushing of the adsorbent to obtain an entirely particulate-free adsorbent prior to the product elution step. Adsorbent with larger pores is also shown to provide faster and more efficient elution both in packed and expanded bed modes. PMID- 11386875 TI - Water content in an engineered dermal replacement during permeation of Me2SO solutions using rapid MR imaging. AB - The successful cryopreservation of cell and tissues typically requires the use of specialized solutions containing cryoprotective agents. At room temperature, the introduction of a cryopreservation solution can result in cell damage/death resulting from osmotic stresses and/or biochemical toxicity of the solution. For tissues, the permeation and equilibration of a cryoprotective solution throughout the tissue is important in enhancing the uniformity and consistency of the postthaw viability of the tissue. Magnetic resonance (MR) is a common nondestructive technique that can be used to quantitate the temporal and spatial composition of water and cryoprotective agents in a three-dimensional system. We have applied a recently developed rapid NMR imaging technique to quantify the transport of water in an artificial dermal replacement upon permeation of dimethyl sulfoxide (Me2SO) solutions. Results indicate that the rate of water transport is slower in the presence of Me2SO molecules. Furthermore, the transport is concentration-dependent, suggesting that Me2SO tends to retain bound water molecules in the tissue. Moreover, water transport decreases with decreasing temperature, and the presence of cells tends to increase water transport. PMID- 11386876 TI - Structural and functional stabilization of L-asparaginase via multisubunit immobilization onto highly activated supports. AB - A new protocol for the stabilization of the quaternary structure of multimeric enzymes has been attempted using as model enzyme (tetrameric) L-asparaginase from Escherichia coli. Such strategy is based upon multisubunit covalent immobilization of the enzyme onto activated supports (agarose-glutaraldehyde). Supports activated with different densities of reactive groups were used; the higher the density of groups, the higher the stabilization attained. However, because of the complexity of that enzyme, even the use of the highest densities of reactive groups was not enough to encompass all four subunits in the immobilization process. Therefore, a further chemical intersubunit cross-linking with aldehyde-dextran was pursued; these derivatives displayed a fully stabilized multimeric structure. In fact, boiling the modified enzyme derivative in the presence of sodium dodecyl sulfate and beta-mercaptoethanol did not lead to release of any enzyme subunit into the medium. Such a derivative, prepared under optimal conditions, retained ca. 40% of the intrinsic activity of the free enzyme and was also functionally stabilized, with thermostabilization enhancements of ca. 3 orders of magnitude when compared with its soluble counterpart. This type of derivative may be appropriate for extracorporeal devices in the clinical treatment of acute leukemia and might thus bring about inherent advantages in that all subunits are covalently bound to the support, with a longer half-life and a virtually nil risk of subunit release into the circulating blood stream. PMID- 11386877 TI - Application of an antibody biochip for p53 detection and cancer diagnosis. AB - Detection of the p53 tumor suppressor gene is important in early cancer diagnostics because alterations in the gene have been associated with carcinogenic manifestations in several tissue types in humans. We have developed an antibody-based detection instrument, the biochip, to detect the presence of the anti-p53 antibody in human serum. The design of this highly integrated detector system is based on miniaturized phototransistors having multiple optical sensing elements, amplifiers, discriminators, and logic circuitry on an IC board. The system utilizes laser excitation and fluorescence signals to detect complex formation between the p53 monoclonal antibody and the p53 antigen. Recognition antibodies are immobilized on a nylon membrane platform and incubated in solutions containing antigens labeled with Cy5, a fluorescent cyanine dye. Subsequently, this membrane is placed on the detection platform of the biochip and fluorescence signal is induced using a 632.8-nm He-Ne laser. Using this immuno-biochip, we have been able to detect binding of the p53 monoclonal antibody to the human p53 cancer protein in biological matrices. The performance of the integrated phototransistors and amplifier circuits of the biochip, previously evaluated through measurement of the signal output response for various concentrations of fluorescein-labeled molecules, have illustrated the linearity of the microchip necessary for quantitative analysis. The design of this biochip permits sensitive, selective and direct measurements of a variety of antigen-antibody formations at very low concentrations. Furthermore, the acquisitions of the qualitative and quantitative results are accomplished rapidly, in about 15 min. These features demonstrate the potential of this antibody-based biochip for simple, rapid and early biomedical diagnostics of cancer. PMID- 11386878 TI - Enzymatic computing. AB - The conformational dynamics of enzymes is a computational resource that fuses milieu signals in a nonlinear fashion. Response surface methodology can be used to elicit computational functionality from enzyme dynamics. We constructed a tabletop prototype to implement enzymatic signal processing in a device context and employed it in conjunction with malate dehydrogenase to perform the linearly inseparable exclusive-or operation. This shows that proteins can execute signal processing operations that are more complex than those performed by individual threshold elements. We view the experiments reported, though restricted to the two-variable case, as a stepping stone to computational networks that utilize the precise reproducibility of proteins, and the concomitant reproducibility of their nonlinear dynamics, to implement complex pattern transformations. PMID- 11386879 TI - Sugar metabolic analysis of suspensions of plant cells using an FT-IR/ATR method. AB - A simple, rapid and accurate evaluation of the sugar uptake rate of suspended plant cells from culture media was developed with the predicted sugar contents measured by mid-infrared spectroscopy using a Fourier transform infrared (FT-IR) spectrometer equipped with an attenuated total reflectance (ATR) accessory. We performed plant cell cultivation with Nicotiana tabacum cv. Bright Yellow No.2 (TBY-2) in culture media, which had various combinations of glucose, fructose and sucrose concentrations at the initial stage, and measured simultaneously each sugar content in the medium by the FT-IR/ATR method. By applying a logistic function to the predicted sugar contents and cell density in the medium during cultivation, the specific sugar uptake rates by the suspended TBY-2 cells were easily and continuously obtained. Thus the kinetic sugar uptake phenomena by the TBY-2 cells were well confirmed overall using the developed method. Additionally it was found that the fraction of sucrose of the initial total sugar content might kinetically affect the sugar uptake process and cell growth. Also, the relationship between the nondimensional cell density and sucrose content could be classified into three groups on the basis of the initial fraction of sucrose. PMID- 11386880 TI - Stochastic modeling of affinity adsorption. AB - A stochastic model is described that allows surface proximity and packing effects to be incorporated into predictions of adsorption kinetics and equilibrium of affinity adsorption. Equilibrium predictions show that, depending on conditions chosen, the results obtained for equilibrium conditions can exhibit either a Freundlich- or a Langmuir-type relationship. Under conditions of surface density imposed adsorption constraints, the time taken for equilibrium to be reached increases as the "off" constant is decreased. This suggests that for resins having a high immobilized ligand density binding kinetics may be more highly limited by the "off" constant than by mass transfer limitations. PMID- 11386881 TI - Raspberry ketone from submerged cultured cells of the basidiomycete Nidula niveo tomentosa. AB - The basidiomycete Nidula niveo-tomentosa produced 4-(4-hydroxyphenyl)-butan-2-one (raspberry ketone), one of the character impact components of raspberry flavor, and its corresponding alcohol. A systematic attempt was made to improve the productivity of this fungus. Variation of nutrient medium composition, precursor amount, time of supplementation, and cultivation period yielded a 50-fold increase in metabolite concentrations. Raspberry ketone and alcohol were easily isolated from the culture medium by solvent extraction. Glycosidically bound forms or accumulation of raspberry compounds in fungal cells were not detected. This microbial process offers an alternative for the production of natural raspberry flavor. PMID- 11386882 TI - Programmed Escherichia coli cell lysis by expression of cloned T4 phage lysis genes. AB - Self-disruptive Escherichia coli that produces foreign target protein was developed. E. coli was co-transformed with two vector plasmids, a target gene expression vector and a lysis gene expression vector. The lytic protein was produced after the expression of the target gene, resulting in simplification of the cell disruption process. In this study, the expression of cloned T4 phage gene e or t was used for the disruption of E. coli that produced beta glucuronidase (GUS) as a model target protein. The expression of gene e did not lead to prompt cell disruption but weakened the cell wall. Resuspension with deionized water facilitated cell lysis, and GUS activity was observed in the resuspended liquid. Expression of gene e at mid logarithmic growth phase was the optimal induction period for GUS production and release. On the other hand, the expression of gene t induced immediate cell lysis, and intracellular GUS was released to the culture medium. Maximum GUS production was obtained when gene t was induced at late logarithmic growth phase. PMID- 11386884 TI - Unfolding and conformational distributions during protein precipitation. AB - The association of misfolded proteins, or aggregation, is a critical problem in a number of human diseases as well during the expression, refolding, formulation, and delivery of therapeutic proteins. In this study, we investigate lysozyme precipitation with hydrogen exhange using nuclear magnetic resonance (NMR) and mass spectrometry (MS). We show that MS can reveal the presence of conformational distributions, albeit without the detailed structural information afforded by NMR. Further, we find that increases in precipitant concentration alter the structure and composition of precipitates. The selective unfolding of one portion of the protein in these precipitates is correlated with hydrogen exchange patterns observed under nonprecipitating conditions and in other studies of lysozyme. PMID- 11386883 TI - Model process for removal of caseins from milk of transgenic animals. AB - We describe a method for selective removal of caseins from milk. The method was developed as a model for transgenic milk processing. Raw cow milk spiked with nonmilk proteins was chosen as the model to resemble transgenic animal milk containing recombinant proteins. The most important elements of the process are (1) "deconstruction" of casein micelles in milk by destroying their Ca(2+) core using a chelating agent (EDTA), thus freeing any protein that might be entrapped in casein aggregates, and (2) "reconstruction" of micelles by providing them with a new Ca(2+) core, thus precipitating them away from the whey proteins, and the protein of interest. Calcium phosphate particles (CAP) were used to reform the disrupted casein micelles. The crystal clear supernatant fraction generated by this method provided >90% recovery and 6- to 13-fold concentration of the desired protein. Product-rich supernatant contained no detectable casein residues, as silver-stained SDS-PAGE and Western blot analyses demonstrated. PMID- 11386886 TI - Non-high-density lipoprotein cholesterol level as potential risk predictor and therapy target. PMID- 11386887 TI - What killed Mozart? PMID- 11386888 TI - Endoscopy for acute nonvariceal upper gastrointestinal tract hemorrhage: is sooner better? A systematic review. AB - BACKGROUND: While the effectiveness of upper endoscopy has been established for acute nonvariceal upper gastrointestinal tract hemorrhage, its optimal timing has not been clearly defined. Early endoscopy has been advocated for its ability to achieve prompt diagnosis, risk stratification, and therapeutic hemostasis. OBJECTIVE: To determine whether early vs delayed endoscopy improves patient and economic outcomes for all risk groups with nonvariceal upper gastrointestinal tract hemorrhage. METHODS: A systematic review of 3 computerized databases (MEDLINE, HEALTHSTAR, and Cochrane Database of Systematic Reviews) was performed along with hand searching of published abstracts to identify English-language citations from 1980 to 2000. RESULTS: Twenty-three studies met explicit inclusion criteria. The highest-quality study examining outcomes in low-risk patients found no significant complications at 1-month follow-up for any outpatients managed with early endoscopy. The largest randomized trial of high-risk patients showed no mortality benefit but a significant decrease in transfusion requirements with early endoscopy. Seven of the 8 studies examining the effect of early endoscopy on length of stay as a measure of resource utilization demonstrated a significant reduction compared with that of delayed endoscopy. However, most included studies were found to suffer from 1 or more potentially significant methodologic shortcomings. CONCLUSIONS: The overwhelming majority of existing data suggest that early endoscopy is safe and effective for all risk groups. The clinical and economic outcomes of early endoscopy should be confirmed in additional well designed randomized controlled trials. Given the strength of the evidence, efforts to develop a more standardized and time-sensitive approach to acute nonvariceal upper gastrointestinal tract hemorrhage should be undertaken. PMID- 11386889 TI - Use of statins and the subsequent development of deep vein thrombosis. AB - BACKGROUND: Some of the benefit of statins for the prevention of cardiovascular disease may be due to their antithrombotic properties. Little is known about the effect of these drugs on the development of deep vein thrombosis. MATERIALS AND METHODS: We conducted a retrospective cohort study over an 8-year period by linking Ontario provincial health care administrative databases covering more than 1.4 million Ontario residents aged 65 years or older. We excluded those with a documented history of atherosclerosis, venous thromboembolism, or cancer within 36 months prior to study enrollment, as well as those prescribed warfarin sodium within 12 months before enrollment. In the primary cohort, we evaluated the subsequent risk of deep vein thrombosis (DVT) among men and women prescribed thyroid replacement therapy, nonstatin lipid-lowering agents, or statins. A second cohort of women only was evaluated in a similar fashion, but estrogen use was added as a third comparison drug group. RESULTS: There were 125 862 men and women in the primary cohort. After adjusting for age; sex; prior hospitalization; newly diagnosed cancer; or prescribed aspirin, warfarin, or estrogen, statin users (n = 77 993) had an associated decreased risk of DVT relative to those prescribed thyroid replacement therapy (n = 35 978) (adjusted hazard ratio [HR], 0.78; 95% confidence interval [CI], 0.69-0.87). Compared with thyroid replacement therapy, users of nonstatin lipid-lowering agents (n = 11 891) did not seem to be at lower risk for deep vein thrombosis (HR, 0.97; 95% CI, 0.79-1.18). In the secondary cohort of 89 508 women, after adjusting for age, prior hospitalization, newly diagnosed cancer, or prescribed aspirin or warfarin, estrogen users (n = 29 165) had an associated increased risk for DVT compared with those receiving thyroid replacement therapy (n = 22 118) (HR, 1.16; 95% CI, 1.01-1.33), while statin users had an associated decreased risk (HR, 0.68; 95% CI, 0.59-0.79). Nonstatin lipid-lowering agents (n = 5155) were not associated with a reduced risk of DVT compared with thyroid replacement therapy (HR, 0.84; 95% CI, 0.63 1.12). CONCLUSION: Among selected individuals aged 65 years or older, statins were associated with a 22% relative risk reduction in the risk of DVT. A randomized clinical trial is needed to evaluate the efficacy of statins for the primary and secondary prevention of DVT. PMID- 11386890 TI - Non-high-density lipoprotein cholesterol level as a predictor of cardiovascular disease mortality. AB - BACKGROUND: Non-high-density lipoprotein cholesterol (non-HDL-C) contains all known and potential atherogenic lipid particles. Therefore, non-HDL-C level may be as good a potential predictor of risk for cardiovascular disease (CVD) as low density lipoprotein cholesterol (LDL-C). OBJECTIVES: To determine whether non-HDL C level could be useful in predicting CVD mortality and to compare the predictive value of non-HDL-C and LDL-C levels. METHODS: Data are from the Lipid Research Clinics Program Follow-up Study, a mortality study with baseline data gathered from 1972 through 1976, and mortality ascertained through 1995. A total of 2406 men and 2056 women aged 40 to 64 years at entry were observed for an average of 19 years, with CVD death as the main outcome measure. RESULTS: A total of 234 CVD deaths in men and 113 CVD deaths in women occurred during follow-up. Levels of HDL-C and non-HDL-C at baseline were significant and strong predictors of CVD death in both sexes. In contrast, LDL-C level was a somewhat weaker predictor of CVD death in both. Differences of 0.78 mmol/L (30 mg/dL) in non-HDL-C and LDL-C levels corresponded to increases in CVD risk of 19% and 15%, respectively, in men. In women, differences of 0.78 mmol/L (30 mg/dL) in non-HDL-C and LDL-C levels corresponded to increases in CVD risk of 11% and 8%, respectively. CONCLUSIONS: Non-HDL-C level is a somewhat better predictor of CVD mortality than LDL-C level. Screening for non-HDL-C level may be useful for CVD risk assessment. PMID- 11386891 TI - Consequences of asymptomatic bacteriuria in women with diabetes mellitus. AB - BACKGROUND: Women with diabetes mellitus (DM) have asymptomatic bacteriuria (ASB) more often than women without DM. It is unknown, however, what the consequences of ASB are in these women. OBJECTIVE: To compare women with DM with and without ASB for the development of symptomatic urinary tract infections (UTIs), renal function, and secondary complications of DM during an 18-month follow-up period. METHODS: In this multicenter study we monitored women with DM with and without ASB for the development of symptomatic UTIs, renal function, and secondary complications (ie, retinopathy, neuropathy, microvascular, or macrovascular diseases). Data on the first 18-month follow-up period are presented. RESULTS: At least 1 uncontaminated urine culture was available from 636 women (258 with type 1 DM and 378 with type 2 DM). The prevalence of ASB at baseline was 26% (21% for those with type 1 DM and 29% for those with type 2 DM). Follow-up results were available for 589 (93%) of the 636 women. Of these 589 women, 115 (20%) (14% with type 1 DM and 23% with type 2 DM) developed a symptomatic UTI. Women with type 2 DM and ASB at baseline had an increased risk of developing a UTI during the 18 month follow-up (19% without ASB vs 34% with ASB, P =.006). In contrast, there was no difference in the incidence of symptomatic UTI between women with type 1 DM and ASB and those without ASB (12% with ASB vs 15% without ASB). However, women with type 1 DM and ASB had a tendency to have a faster decline in renal function than those without ASB (relative increase in serum creatinine level 4.6% vs 1.5%, P = 0.2). CONCLUSION: Women with type 2 DM and ASB have an increased risk of developing a symptomatic UTI than those without ASB. PMID- 11386892 TI - Echocardiographic examination of women previously treated with fenfluramine: long term follow-up of a randomized, double-blind, placebo-controlled trial. AB - BACKGROUND: Fenfluramine hydrochloride was withdrawn from the market in September 1997 after reports of heart valve abnormalities in patients who used it. The prevalence of echocardiographic abnormalities and the clinical cardiovascular status of patients who received fenfluramine monotherapy remains uncertain. METHODS: A long-term, follow-up evaluation was undertaken in subjects who were randomly assigned to receive either fenfluramine hydrochloride (60 mg daily) or placebo as part of a double-blind smoking cessation therapy study. Cardiovascular status was evaluated by echocardiography, medical history, and physical examination. RESULTS: From the group of 720 smokers who had originally participated in the smoking cessation therapy trial, 619 women were enrolled; data from 530 (276 in the fenfluramine group and 254 in the placebo group) were evaluable. No statistically significant differences were identified in the prevalence of aortic or mitral regurgitation by Food and Drug Administration criteria or by grade, aortic or mitral valve leaflet mobility restriction or thickening, elevated pulmonary artery systolic pressure, or abnormal left ventricular ejection fraction. No significant differences were demonstrated in cardiovascular status by physical examination, and no serious cardiac events were noted among fenfluramine-treated subjects. CONCLUSION: There was no evidence of drug-related heart disease up to 4.9 years after anorexigen therapy in subjects who were randomly assigned to receive fenfluramine at the recommended dose for up to 3 months. PMID- 11386893 TI - Is patients' perception of time spent with the physician a determinant of ambulatory patient satisfaction? AB - BACKGROUND: Time management in ambulatory patient visits is increasingly critical. Do patients who perceive a longer visit with internists report increased satisfaction? METHODS: Prospective survey of 1486 consecutively encountered ambulatory visits to 16 primary care physicians (PCPs) in an academic primary care clinic. Patients were queried regarding demographics, health status, perception of time spent before and after ambulatory visits, whether the physician appeared rushed, and visit satisfaction. Physicians were queried regarding time spent, estimated patient satisfaction, and whether they felt rushed. RESULTS: In 69% of 1486 consecutive visits, patient previsit expectation of visit duration was 20 minutes or less. Patient and PCP postvisit estimates of time spent significantly exceeded patient previsit time expectation. Patients who estimated that they spent more time than expected with the PCP were significantly more satisfied with the visit. When patient postvisit estimate of time spent was less than the previsit expectation, visit satisfaction was significantly lower independent of time spent. Patient worry about health and lower self-perceived health status were significantly associated with patient expectation for longer visits. Primary care physicians felt rushed in 10% of encounters. Although PCPs estimated patient satisfaction was significantly lower when they felt rushed, patient satisfaction was identical when PCPs did and did not feel rushed. Patients indicated that PCPs appeared rushed in 3% of encounters, but this perception did not affect patient satisfaction. CONCLUSION: Perceived ambulatory visit duration and meeting or exceeding patient expectation of time needed to be spent with the physician are determinants of patient satisfaction in an ambulatory internal medicine practice. PMID- 11386894 TI - Are the results of randomized controlled trials on anticoagulation in patients with atrial fibrillation generalizable to clinical practice? AB - BACKGROUND: Randomized trials demonstrate a clear benefit of anticoagulation in patients with atrial fibrillation at risk of stroke, but the proportion of eligible patients who are treated with anticoagulants remains low. The reluctance to treat all eligible patients with anticoagulants may be due to studies in clinical practice showing variable risk-benefit, raising concerns about application to general medical practice. METHODS: A systematic review of published medical literature was performed to identify studies of patients with atrial fibrillation who were treated with warfarin in actual clinical practice. Data from these studies were compared with pooled data from randomized controlled trials. RESULTS: Three studies met the predefined criteria, each in a different health care setting, totaling 410 patients with 842 patient-years of follow-up. Patients in clinical practice were older and had more comorbid conditions compared with trial participants. However, the ischemic stroke rate was similar between clinical practice and randomized studies (1.8% [95% confidence interval (CI), 0.9%-2.7%] vs 1.4% [95% CI, 0.9%-2.0%]). Intracranial hemorrhage (0.1% [95% CI, 0%-0.3%] vs 0.3% [95% CI, 0.06%-0.5%]) and major bleeding (1.1% [95% CI, 0.4% 1.8%] vs 1.3% [95% CI, 0.8%-1.8%]) rates were also similar. There was a higher rate of minor bleeding in clinical practice than in trials (12.0% [95% CI, 9.7% 14.3%] vs 7.9% [95% CI, 6.6%-9.2%]). CONCLUSIONS: Patients who undergo anticoagulation for atrial fibrillation in actual clinical practice differ from those in randomized trials, but have similar rates of stroke and major bleeding. The risk of minor bleeding is higher and may require more intensive monitoring in practice. PMID- 11386895 TI - Postmenopausal estrogen use, type of menopause, and lens opacities: the Framingham studies. AB - BACKGROUND: Previous studies of estrogen replacement therapy and lens opacities have not reported consistent findings. OBJECTIVE: To investigate whether postmenopausal estrogen use is associated with the occurrence of age-related lens opacities (nuclear, cortical, and posterior subcapsular). METHODS: Surviving members of the original cohort of the Framingham Heart Study who also participated in the Framingham Eye Study (1986-1989) were examined for the absence or presence of lens opacities. Data from the Framingham Heart Study, including information on menopausal status (collected biennially from approximately 1948) and use of estrogen replacement therapy (collected biennially from approximately 1960) were used to examine associations between lens opacities and duration of postmenopausal estrogen use, type of menopause, and age at menopause. Five hundred twenty-nine women, aged 66 to 93 years, were included. Multivariable-adjusted odds ratios of specific types of lens opacities were calculated for (1) duration of estrogen use (never and 1-2, 3-9, and >/=10 years), (2) surgical vs natural menopause, and (3) age at menopause. RESULTS: Longer duration of postmenopausal estrogen therapy was inversely associated with the presence of nuclear lens opacities in an adjusted model. Women who had taken estrogen for 10 years or longer had a 60% reduction in risk compared with nonusers (odds ratio, 0.4; 95% confidence interval, 0.2-1.01). Longer duration of estrogen use was associated with fewer posterior subcapsular opacities at a borderline level of significance. No association was noted for cortical opacities. The risk of posterior subcapsular opacities was significantly increased for women who had undergone surgical menopause compared with women with natural menopause (odds ratio, 2.2; 95% confidence interval, 1.1-4.3). No association was noted for lens opacities and age at menopause. CONCLUSION: Data from our study and other studies suggest that a reduction in the risk of lens opacities may be an additional benefit of postmenopausal estrogen use. PMID- 11386896 TI - The effect of creatine and resistance training on plasma homocysteine concentration in healthy volunteers. PMID- 11386897 TI - Eosinophilic fasciitis and simvastatin. PMID- 11386898 TI - Other factors in self-reported hypertension treatment practices among primary care physicians. PMID- 11386900 TI - Body mass index and mortality among hospitalized elderly patients. PMID- 11386902 TI - Reductase inhibitors and the risk of cancer. PMID- 11386904 TI - A follow-up to "a Jewish physician amidst the holocaust". PMID- 11386905 TI - Appropriate first-line lipid-lowering therapy for type 1 diabetes mellitus. PMID- 11386908 TI - A piece of my mind: the list. PMID- 11386909 TI - Suicide prevention plan calls for physicians' help. PMID- 11386910 TI - Innovative program for mentally ill inmates. PMID- 11386912 TI - From the Food and Drug Administration. PMID- 11386915 TI - Positron emission tomography to evaluate lung lesions. PMID- 11386917 TI - Automobile traffic, atmospheric pollution, and childhood asthma. PMID- 11386919 TI - The role of local institutional review boards in protecting human research subjects. PMID- 11386921 TI - Informing patients of uncertainty in clinical trials. PMID- 11386923 TI - Denial of handgun purchase to violent misdemeanants. PMID- 11386925 TI - Botulinum toxin in biowarfare. PMID- 11386926 TI - Mutation in the SPINK1 trypsin inhibitor gene, alcohol use, and chronic pancreatitis. PMID- 11386927 TI - Effect of ramipril vs amlodipine on renal outcomes in hypertensive nephrosclerosis: a randomized controlled trial. AB - CONTEXT: Incidence of end-stage renal disease due to hypertension has increased in recent decades, but the optimal strategy for treatment of hypertension to prevent renal failure is unknown, especially among African Americans. OBJECTIVE: To compare the effects of an angiotensin-converting enzyme (ACE) inhibitor (ramipril), a dihydropyridine calcium channel blocker (amlodipine), and a beta blocker (metoprolol) on hypertensive renal disease progression. DESIGN, SETTING, AND PARTICIPANTS: Interim analysis of a randomized, double-blind, 3 x 2 factorial trial conducted in 1094 African Americans aged 18 to 70 years with hypertensive renal disease (glomerular filtration rate [GFR] of 20-65 mL/min per 1.73 m(2)) enrolled between February 1995 and September 1998. This report compares the ramipril and amlodipine groups following discontinuation of the amlodipine intervention in September 2000. INTERVENTIONS: Participants were randomly assigned to receive amlodipine, 5 to 10 mg/d (n = 217), ramipril, 2.5 to 10 mg/d (n = 436), or metoprolol, 50 to 200 mg/d (n = 441), with other agents added to achieve 1 of 2 blood pressure goals. MAIN OUTCOME MEASURES: The primary outcome measure was the rate of change in GFR; the main secondary outcome was a composite index of the clinical end points of reduction in GFR of more than 50% or 25 mL/min per 1.73 m(2), end-stage renal disease, or death. RESULTS: Among participants with a urinary protein to creatinine ratio of >0.22 (corresponding approximately to proteinuria of more than 300 mg/d), the ramipril group had a 36% (2.02 [SE, 0.74] mL/min per 1.73 m(2)/y) slower mean decline in GFR over 3 years (P =.006) and a 48% reduced risk of the clinical end points vs the amlodipine group (95% confidence interval [CI], 20%-66%). In the entire cohort, there was no significant difference in mean GFR decline from baseline to 3 years between treatment groups (P =.38). However, compared with the amlodipine group, after adjustment for baseline covariates the ramipril group had a 38% reduced risk of clinical end points (95% CI, 13%-56%), a 36% slower mean decline in GFR after 3 months (P =.002), and less proteinuria (P<.001). CONCLUSION: Ramipril, compared with amlodipine, retards renal disease progression in patients with hypertensive renal disease and proteinuria and may offer benefit to patients without proteinuria. PMID- 11386928 TI - High-density lipoprotein cholesterol and ischemic stroke in the elderly: the Northern Manhattan Stroke Study. AB - CONTEXT: Elevated high-density lipoprotein cholesterol (HDL-C) levels have been shown to be protective against cardiovascular disease. However, the association of specific lipoprotein classes and ischemic stroke has not been well defined, particularly in higher-risk minority populations. OBJECTIVE: To evaluate the association between HDL-C and ischemic stroke in an elderly, racially or ethnically diverse population. DESIGN: Population-based, incident case-control study conducted July 1993 through June 1997. SETTING: A multiethnic community in northern Manhattan, New York, NY. PARTICIPANTS: Cases (n = 539) of first ischemic stroke (67% aged >/=65 years; 55% women; 53% Hispanic, 28% black, and 19% white) were enrolled and matched by age, sex, and race or ethnicity to stroke-free community residents (controls; n = 905). MAIN OUTCOME MEASURE: Independent association of fasting HDL-C levels, determined at enrollment, with ischemic stroke, including atherosclerotic and nonatherosclerotic ischemic stroke subtypes. RESULTS: After risk factor adjustment, a protective effect was observed for HDL-C levels of at least 35 mg/dL (0.91 mmol/L) (odds ratio [OR], 0.53; 95% confidence interval [CI], 0.39-0.72). A dose-response relationship was observed (OR, 0.65; 95% CI, 0.47-0.90 and OR, 0.31; 95% CI, 0.21-0.46) for HDL-C levels of 35 to 49 mg/dL (0.91-1.28 mmol/L) and at least 50 mg/dL (1.29 mmol/L), respectively. The protective effect of a higher HDL-C level was significant among participants aged 75 years or older (OR, 0.51; 95% CI, 0.27-0.94), was more potent for the atherosclerotic stroke subtype (OR, 0.20; 95% CI, 0.08-0.50), and was present in all 3 racial or ethnic groups studied. CONCLUSIONS: Increased HDL C levels are associated with reduced risk of ischemic stroke in the elderly and among different racial or ethnic groups. These data add to the evidence relating lipids to stroke and support HDL-C as an important modifiable stroke risk factor. PMID- 11386929 TI - Mortality and locomotion 6 months after hospitalization for hip fracture: risk factors and risk-adjusted hospital outcomes. AB - CONTEXT: Hip fracture is a common clinical problem that leads to considerable mortality and disability. A need exists for a practical means to monitor and improve outcomes, including function, for patients with hip fracture. OBJECTIVES: To identify and compare the importance of significant prefracture predictors of functional status and mortality at 6 months for patients hospitalized with hip fracture and to compare risk-adjusted outcomes for hospitals providing initial care. DESIGN: Prospective study with data obtained from medical records and through structured interviews with patients and proxies. SETTING AND PARTICIPANTS: A total of 571 adults aged 50 years or older with hip fracture who were admitted to 4 New York, NY, metropolitan hospitals between August 1997 and August 1998. MAIN OUTCOME MEASURES: In-hospital and 6-month mortality; locomotion at 6 months; and adverse outcomes at 6 months, defined as death or needing assistance to ambulate, compared by hospital, adjusting for patient risk factors. RESULTS: The in-hospital mortality rate was 1.6%. At 6 months, the mortality rate was 13.5%, and another 12.8% needed total assistance to ambulate. Laboratory values were strong predictors of mortality but were not significantly associated with locomotion. Age and prefracture residence at a nursing home were significant predictors of locomotion (P =.02 for both) but were not significantly associated with mortality. Adjustment for baseline characteristics either substantially augmented or diminished interhospital differences in outcomes. Two hospitals had 1 outcome (functional status or mortality) that was significantly worse than the overall mean while the other outcome was nonsignificantly better than average. CONCLUSIONS: Mortality and functional status ideally should be considered both together and individually to distinguish effects limited to one or the other outcome. Hospital performance for these 2 measures may differ substantially after adjustment, probably because different processes of care are important to each outcome. PMID- 11386930 TI - Enzyme replacement therapy in Fabry disease: a randomized controlled trial. AB - CONTEXT: Fabry disease is a metabolic disorder without a specific treatment, caused by a deficiency of the lysosomal enzyme alpha-galactosidase A (alpha-gal A). Most patients experience debilitating neuropathic pain and premature mortality because of renal failure, cardiovascular disease, or cerebrovascular disease. OBJECTIVE: To evaluate the safety and efficacy of intravenous alpha-gal A for Fabry disease. DESIGN AND SETTING: Double-blind placebo-controlled trial conducted from December 1998 to August 1999 at the Clinical Research Center of the National Institutes of Health. PATIENTS: Twenty-six hemizygous male patients, aged 18 years or older, with Fabry disease that was confirmed by alpha-gal A assay. INTERVENTION: A dosage of 0.2 mg/kg of alpha-gal A, administered intravenously every other week (12 doses total). MAIN OUTCOME MEASURE: Effect of therapy on neuropathic pain while without neuropathic pain medications measured by question 3 of the Brief Pain Inventory (BPI). RESULTS: Mean (SE) BPI neuropathic pain severity score declined from 6.2 (0.46) to 4.3 (0.73) in patients treated with alpha-gal A vs no significant change in the placebo group (P =.02). Pain-related quality of life declined from 3.2 (0.55) to 2.1 (0.56) for patients receiving alpha-gal A vs 4.8 (0.59) to 4.2 (0.74) for placebo (P =.05). In the kidney, glomeruli with mesangial widening decreased by a mean of 12.5% for patients receiving alpha-gal vs a 16.5% increase for placebo (P =.01). Mean inulin clearance decreased by 6.2 mL/min for patients receiving alpha-gal A vs 19.5 mL/min for placebo (P =.19). Mean creatinine clearance increased by 2.1 mL/min (0.4 mL/s) for patients receiving alpha-gal A vs a decrease of 16.1 mL/min (0.3 mL/s) for placebo (P =.02). In patients treated with alpha-gal A, there was an approximately 50% reduction in plasma glycosphingolipid levels, a significant improvement in cardiac conduction, and a significant increase in body weight. CONCLUSION: Intravenous infusions of alpha-gal A are safe and have widespread therapeutic efficacy in Fabry disease. PMID- 11386931 TI - Cancer screening in elderly patients: a framework for individualized decision making. AB - Considerable uncertainty exists about the use of cancer screening tests in older people, as illustrated by the different age cutoffs recommended by various guideline panels. We suggest that a framework to guide individualized cancer screening decisions in older patients may be more useful to the practicing clinician than age guidelines. Like many medical decisions, cancer screening decisions require weighing quantitative information, such as risk of cancer death and likelihood of beneficial and adverse screening outcomes, as well as qualitative factors, such as individual patients' values and preferences. Our framework first anchors decisions through quantitative estimates of life expectancy, risk of cancer death, and screening outcomes based on published data. Potential benefits of screening are presented as the number needed to screen to prevent 1 cancer-specific death, based on the estimated life expectancy during which a patient will be screened. Estimates reveal substantial variability in the likelihood of benefit for patients of similar ages with varying life expectancies. In fact, patients with life expectancies of less than 5 years are unlikely to derive any survival benefit from cancer screening. We also consider the likelihood of potential harm from screening according to patient factors and test characteristics. Some of the greatest harms of screening occur by detecting cancers that would never have become clinically significant. This becomes more likely as life expectancy decreases. Finally, since many cancer screening decisions in older adults cannot be answered solely by quantitative estimates of benefits and harms, considering the estimated outcomes according to the patient's own values and preferences is the final step for making informed screening decisions. PMID- 11386932 TI - Stroke in a healthy 46-year-old man. AB - This article presents the case of a healthy 46-year-old man who experienced a dissection of the internal carotid artery. The diagnosis of this condition is not usually clear-cut, especially in a young patient with unremarkable medical history, and because of the similarity of symptoms with migraine. Often there is no obvious cause of a cerebral artery dissection, although subtle abnormalities of connective tissue may be present. Anticoagulation is generally used for therapy, but clinical trials are lacking. Carotid artery dissection should be considered as a cause of stroke in young healthy adults. PMID- 11386933 TI - Tularemia as a biological weapon: medical and public health management. AB - OBJECTIVE: The Working Group on Civilian Biodefense has developed consensus-based recommendations for measures to be taken by medical and public health professionals if tularemia is used as a biological weapon against a civilian population. PARTICIPANTS: The working group included 25 representatives from academic medical centers, civilian and military governmental agencies, and other public health and emergency management institutions and agencies. EVIDENCE: MEDLINE databases were searched from January 1966 to October 2000, using the Medical Subject Headings Francisella tularensis, Pasteurella tularensis, biological weapon, biological terrorism, bioterrorism, biological warfare, and biowarfare. Review of these references led to identification of relevant materials published prior to 1966. In addition, participants identified other references and sources. CONSENSUS PROCESS: Three formal drafts of the statement that synthesized information obtained in the formal evidence-gathering process were reviewed by members of the working group. Consensus was achieved on the final draft. CONCLUSIONS: A weapon using airborne tularemia would likely result 3 to 5 days later in an outbreak of acute, undifferentiated febrile illness with incipient pneumonia, pleuritis, and hilar lymphadenopathy. Specific epidemiological, clinical, and microbiological findings should lead to early suspicion of intentional tularemia in an alert health system; laboratory confirmation of agent could be delayed. Without treatment, the clinical course could progress to respiratory failure, shock, and death. Prompt treatment with streptomycin, gentamicin, doxycycline, or ciprofloxacin is recommended. Prophylactic use of doxycycline or ciprofloxacin may be useful in the early postexposure period. PMID- 11386934 TI - Selection of antihypertensive therapy for patients with hypertensive renal disease. PMID- 11386935 TI - Informed choice in cancer screening. PMID- 11386937 TI - msJAMA: residency selection: making a match. PMID- 11386938 TI - msJAMA: from medical student to intern: where are the role models? PMID- 11386939 TI - msJAMA: is medical school the right place to choose a specialty? PMID- 11386940 TI - msJAMA: residency selection process and the match: does anyone believe anybody? PMID- 11386947 TI - Systematic reviews: a critical first step. PMID- 11386948 TI - Can we train a lifelong learner? PMID- 11386949 TI - Evidence-based medicine journal club: what's it all about and what's in it for me? PMID- 11386950 TI - Steroids for otitis media with effusion: a systematic review. AB - BACKGROUND: Otitis media with effusion (OME) is common and may cause hearing loss with associated delayed language development in children. Treatment remains controversial. OBJECTIVE: To examine evidence for or against treating OME with systemic or topical nasal steroids. DATA SOURCES: We searched the Cochrane Controlled Trials Register using the terms otitis media; otitis media with effusion; glue ear; or OME and steroids; glucocorticoids; glucocorticoids, synthetic; glucocorticoids, topical; or anti-inflammatory agents, steroidal; or various combinations of these terms. EMBASE and MEDLINE were also searched. STUDY SELECTION: Randomized controlled trials of oral and topical nasal steroids, either alone or in combination with another agent such as an antibiotic, were included. Ten studies met the inclusion criteria. DATA EXTRACTION: Data extraction and methodological quality assessment were performed by the 2 of us (C.C.B. and J.H.v.d.V.) independently, using standardized methods described in the Cochrane Collaboration Handbook. DATA SYNTHESIS: The odds ratio for OME persisting after short-term follow-up in children treated with oral steroids compared with a control was 0.22 (95% confidence interval, 0.08 = 0.63), and was 0.32 (95% confidence interval, 0.20 = 0.52) for children treated with oral steroids plus an antibiotic compared with a control plus an antibiotic. Trends favored steroids for most other comparisons, but confidence intervals included unity. Trends favored steroids for most other comparisons, but confidence intervals included unity. CONCLUSIONS: Steroids alone or combined with an antibiotic lead to a quicker resolution of OME in the short-term. However, there is no evidence for a long-term benefit from treating hearing loss associated with OME with either oral or topical nasal steroids. These treatments are, therefore, not recommended. PMID- 11386951 TI - Access to care for children of the working poor. AB - CONTEXT: Recent evidence suggests that children in working poor families lack health resources, placing them at risk for inadequate access to care. OBJECTIVES: To examine financial and nonfinancial access and utilization of health services among children in working poor families, and to compare these data with those of children from both nonworking poor and moderate to affluent families. DESIGN: A cross-sectional study of 13 785 children younger than 18 years. PARTICIPANTS: Subjects from the 1997 National Health Interview Survey. MAIN OUTCOME MEASURES: Prevalence and continuity of health insurance coverage, of delayed or missed care, and of unmet care needs; presence and type of usual source of care; and the amount of visits to physicians, emergency departments, and hospitals. RESULTS: Compared with children of nonworking poor parents and moderate to affluent children, more working poor children were uninsured (22% vs 12% and 5%, respectively; P<.01) and experienced disruptions in insurance coverage (P<.01). After adjusting for other covariates, disparities in insurance coverage and continuity persisted, as did delays in care and unmet care needs; these delays were far higher for the working poor. Although these children had access to a regular source of care and had utilization rates comparable with those of other poor children, they differed markedly from moderate to affluent children on structural access and utilization (adjusted odds ratios, 1.5-3.4). CONCLUSIONS: Children in working poor families experience far more barriers to care than other children. Health insurance expansions through the Children's Health Insurance Program and Medicaid, which reduce financial and nonfinancial barriers to care, may help correct these disparities. PMID- 11386952 TI - "Love our kids, lock your guns": a community-based firearm safety counseling and gun lock distribution program. AB - BACKGROUND: Safer storage practices may reduce injury rates by limiting youth access to firearms. OBJECTIVE: To determine if a firearm safety counseling and gun lock distribution program improved storage practices. DESIGN: Community-based before-after trial. SETTING: Urban county in central North Carolina. PARTICIPANTS: One hundred twelve adult gun owners recruited through a mass media advertising campaign. INTERVENTION: In the parking lot of a shopping mall, participants completed a survey, and were then provided with tailored counseling, gun safety information, a gun lock, and instructions to use it. MAIN OUTCOME MEASURES: Firearm storage practices, assessed by survey and personal interview (baseline) and telephone interview (6-month follow-up). RESULTS: Most participants were white (62%), men (63%), had children (58%), and owned a gun for protection (74%). At follow-up, of the 82 participants, 63 (77%) (up from 39 [48%]) reported storing their gun(s) in a locked compartment (P =.004), 59 (72%) (up from 0) reported using gun locks (P =.001), 61 (74%) (up from 57 [69%]) reported storing their ammunition locked in a separate location, 59 (72%) (up from 52 [63%]) reported storing their gun(s) unloaded, and 6 (7%) (down from 15 [18%]) reported storing firearms unlocked and loaded. Participants with children were more likely at baseline to store weapons unlocked and loaded (38 [59%] vs 19 [41%]; P =.02) but were more likely after counseling to lock their weapons (29 [58%] vs 14 [44%]) and remove guns from the home (5 [10%] vs 0 [0%]). CONCLUSIONS: This program prompted reporting of safer firearm storage practices, particularly among parents. Longer follow-up, verification of self-reports and correct use, testing of gun locks, and monitoring firearm injury rates after distribution programs are needed to establish the public health potential of this approach. PMID- 11386953 TI - Use of intravenous methohexital as a sedative in pediatric emergency departments. AB - OBJECTIVE: To evaluate the effectiveness and safety of intravenous methohexital as a sedative in children undergoing emergency computed tomographic scans. DESIGN: Case series. SETTING: An urban pediatric emergency department at a level I trauma center. PARTICIPANTS: Patients receiving intravenous methohexital for sedation to undergo emergency computed tomographic scans. METHODS: Descriptive data were generated, including demographics, doses administered, times of sedation, outcomes, and complications. RESULTS: Data are reported from a total of 55 patients. The doses administered ranged from 0.5 mg/kg to 2.0 mg/kg (mean +/- SD, 1 +/- 0.5 mg/kg). Onset of sedation was rapid (mean +/- SD, 1 +/- 0.4 minutes), sedation was brief (mean +/- SD, 12 +/- 5 minutes), and the mean +/- SD length of the drug's effects was 14 +/- 6 minutes. Sedation was effective in most cases, and only a few patients had complications. CONCLUSION: Intravenous methohexital is a short-acting and effective sedative for use in pediatric emergency departments to obtain computed tomographic scans. PMID- 11386954 TI - Impact of problem-based learning on residents' self-directed learning. AB - OBJECTIVE: To examine the effect of a problem-based learning (PBL) curriculum on self-directed learning behaviors among a group of pediatric residents. METHODS: A controlled comparison study was conducted with 80 pediatric residents at a large urban academic medical center. Residents were observed over 3 distinct but consecutive periods. First, all residents participated in a 3-month-long daily lecture series (pre-exposure phase). Then, for another 3 months, 39 residents (PBL group) were exposed to twice-weekly PBL sessions while 41 residents continued with the daily lectures (lecture-based group) and served as controls. Problem-based learning was withdrawn after 3 months and all residents returned to the lecture series (follow-up phase). Residents' self-directed learning behaviors were assessed through self-administered questionnaires during the pre-exposure, exposure, and follow-up phases. RESULTS: There were no significant preexposure differences in self-directed learning behaviors between the groups. During the exposure phase, the PBL group had significantly higher self-directed learning: 5 or more hours of independent study (26% vs 7%) [corrected] (P=.001); 5 or more hours of medical discussions (28% vs 4%) (P=.008); 2 or more computer literature searches (51% vs 30%) (P=.005); and total hours of self-study per week (6 vs 4 hours) (P<.05). At the 3-month follow-up, the PBL group had returned to baseline levels of self-directed learning and there were no significant differences between the groups. CONCLUSION: Residents exposed to PBL engaged in significantly higher levels of self-directed learning than their counterparts. PMID- 11386955 TI - Does problem-based learning improve residents' self-directed learning? PMID- 11386956 TI - Self-obtained vaginal swabs for diagnosis of treatable sexually transmitted diseases in adolescent girls. AB - OBJECTIVE: To ascertain the acceptability of testing and prevalence of 3 readily treatable sexually transmitted diseases (STDs) (infections with Neisseria gonorrhoeae, Chlamydia trachomatis, and Trichomonas vaginalis) with the use of patient-obtained vaginal swabs. STUDY DESIGN: Study participants at each initial session were asked to provide self-obtained vaginal swabs for ligase chain reaction testing to detect N gonorrhoeae and C trachomatis, and for culture of T vaginalis. SETTING: Behavioral intervention sessions with African American adolescent girls in a nonclinical program to reduce risk of STDs, human immunodeficiency virus infection, and pregnancy. RESULTS: All study participants were offered their choice of STD screening in the context of a traditional pelvic examination or using self-obtained vaginal swabs. All eligible participants chose self-administered vaginal swabs. Of the 512 participants examined at their initial study visit, 28.7% were found to be infected with 1 or more treatable STDs (5.3% with N gonorrhoeae, 17.8% with C trachomatis, and 12.9% with T vaginalis). CONCLUSIONS: With the use of newer detection systems, STDs can be readily detected in nonclinical settings with the use of self-obtained vaginal swabs, providing new opportunities for efforts to control STDs. PMID- 11386957 TI - Prophylaxis against possible human immunodeficiency virus exposure after nonoccupational needlestick injuries or sexual assaults in children and adolescents. AB - BACKGROUND: Nonoccupational human immunodeficiency virus (HIV) postexposure prophylaxis (PEP) for adults has been described, although the Centers for Disease Control and Prevention, Atlanta, Ga, offer no specific recommendations. There is limited information about its use in children and adolescents. OBJECTIVE: To describe the current practices of physicians in pediatric infectious disease (PID) and pediatric emergency medicine (PEM) departments regarding nonoccupational HIV PEP for children and adolescents. DESIGN: Survey. PARTICIPANTS: Directors of all PID and PEM departments with fellowship programs in the United States and Canada between July and November 1998. MAIN OUTCOME MEASURES: General questions regarding HIV PEP and questions concerning 2 scenarios (5-year-old with a needlestick injury and a 15-year-old after sexual assault). RESULTS: The return rate was 67 (78%) of 86 for PID and 36 (75%) of 48 for PEM physicians. Fewer than 20% of physicians reported institutional policies for nonoccupational HIV PEP; 33% had ever initiated nonoccupational HIV PEP. In both scenarios, PID physicians were more likely than PEM physicians to recommend or offer HIV PEP in the first 24 hours after the incident (55 [83%] of 66 vs 20 [56%] of 36 for needlestick injuries [odds ratio, 4.0; 95% confidence interval, 1.6-10.1] and 47 [72%] of 65 vs 16 [50%] of 32 for sexual assault [odds ratio, 2.6; 95% confidence interval, 1.1-6.3]). Seven different antiretroviral agents in single, dual, or triple drug regimens administered for 2 to 12 weeks were suggested. CONCLUSIONS: Although few physicians reported institutional policies, and only one third had ever initiated HIV PEP, many would offer or recommend HIV PEP for children and adolescents within 24 hours after possible HIV exposure. A wide variation of regimens have been suggested. There is a need for a national consensus for nonoccupational HIV PEP. PMID- 11386958 TI - Taste test: children rate flavoring agents used with activated charcoal. AB - OBJECTIVE: To compare flavoring agents added to activated charcoal (AC) to determine which mixture is most palatable to children. DESIGN: Healthy volunteers between the ages of 3 and 17 years participated in a prospective masked trial. Five identical pitchers were prepared containing AC alone, AC with chocolate milk, AC with Coca-Cola (Coca-Cola Corp, Atlanta, Ga), AC with cherry-flavored syrup, and AC with sorbitol. Subjects tasted all 5 substances in random order. Children younger than 8 years rated taste on a 10-point Faces Scale. Children 8 years and older used a 100-point visual analog scale to rate taste and, separately, ease of swallowing. All children were asked which mixture was best. Ratings were compared using 1-way analysis of variance, and comparisons for all pairs were made using the Tukey test. P<.05 was considered significant. RESULTS: Mean age among the 53 children enrolled was 8.3 years; 23 children were younger than 8 years. Girls made up 52% of the group. Taste scores for chocolate milk, Coca-Cola, and cherry-flavored syrup were significantly better than those for no flavoring agent. The scores for ease of swallowing for Coca-Cola, chocolate milk, and cherry-flavored syrup were significantly better than those for either no flavoring agent or sorbitol. When asked to choose a single best flavoring agent, 39% chose chocolate milk, 23% picked Coca-Cola, and 23% chose cherry-flavored syrup. CONCLUSION: The addition of chocolate milk, Coca-Cola, or cherry-flavored syrup to AC improves palatability for children and is favored over no flavoring agent or sorbitol. PMID- 11386959 TI - Performance of a predictive model for streptococcal pharyngitis in children. AB - CONTEXT: Group A beta-hemolytic streptococcus (GABHS) pharyngitis is a common childhood illness. The clinical diagnosis is difficult to determine and laboratory tests have limitations; hence, the condition is generally overdiagnosed and overtreated. Several clinical pediatric-specific predictive models have been published but none have been prospectively studied. OBJECTIVE: To test the performance of a previously published predictive model for GABHS pharyngitis in children in different clinical settings and during different seasons. DESIGN: Prospective cohort study. SETTINGS: Pediatric emergency department and 2 pediatric outpatient clinics. PATIENTS: Children aged between 1 and 18 years with pharyngitis on initial examination at study sites between April 1, 1999, and March 31, 2000. INTERVENTIONS: Recording of clinical features during initial evaluation using a standardized form and recovery of GABHS from patients' throats using reference standard methods. MAIN OUTCOME MEASURES: Posttest probability for GABHS positive throat culture associated with the model's positive predictors (moderate to severe tonsillar swelling, cervical lymphadenopathy [moderate to severe tenderness and enlargement of cervical lymph nodes], scarletiniform rash, and the absence of coryza) and the models' negative predictors (absence of the above signs and the presence of coryza). RESULTS: Of 587 patients analyzed, 218 (37%) had a positive throat culture for GABHS. Forty nine percent were boys. Mean +/- SD age was 6.7 +/- 3.9 years. There was no difference between the subsets within the sample. The posttest probability values for a positive throat culture associated with positive and negative predictors of the model were 79% and 12%, respectively. CONCLUSIONS: A pediatric predictive model for GABHS pharyngitis performed better than physicians' subjective estimates for a positive throat culture and was comparable with a rapid antigen detection test. The model performed consistently well in different populations and across seasons. It can be useful if reliable microbiological testing and/or follow-up are not attainable. PMID- 11386960 TI - Parent advisory groups in pediatric practices: parents' and professionals' perceptions. AB - OBJECTIVE: To describe the benefits perceived by parents and professionals from their participation in parent advisory groups (PAGs). DESIGN: Retrospective telephone survey. SETTING: Parent advisory groups were established in 4 community based pediatric practices. Each group included parents of children with chronic health conditions, health care providers, and a Department of Public Health staff member. PARTICIPANTS: The mothers and fathers of children with chronic health conditions completed a telephone survey. These parents also were enrolled in a broader intervention aimed at providing primary care for children with chronic conditions. INTERVENTION: Groups met regularly in the pediatric office setting. Logistical arrangements and activities varied among the 4 groups. MAIN OUTCOME MEASURES: Parents and professionals rated their perceptions of the PAGs based on a structured list of potential benefits. RESULTS: Mothers viewed the PAGs as most beneficial in diminishing their isolation, helping them to understand family concerns, increasing their influence on the pediatric care provided to their children, and improving their knowledge of community resources. Professionals believed that the PAGs enhanced their awareness of common family needs, increased their skills at collaborating with families, and helped them to feel more effective. All professionals and most parents were interested in continuing their involvement in a PAG. CONCLUSIONS: Parent advisory groups may benefit families and professionals who care for them by (1) fostering collaborative relationships and communication, (2) increasing a sense of social support among families, (3) increasing families' knowledge of community-based resources, and (4) increasing the families' sense of efficacy and involvement in the care of their own and others' children. PMID- 11386961 TI - Dental concerns unrelated to trauma in the pediatric emergency department: barriers to care. AB - OBJECTIVES: To describe patients with nontraumatic dental problems treated in our pediatric emergency department (PED) and to determine if barriers to access prompted seeking care in the PED rather than from a dentist or dental clinic. DESIGN: Questionnaire administered to a convenience sample of patients with nontraumatic dental complaints. SETTING: An urban PED. MAIN OUTCOME MEASURES: Insurance status, primary medical and dental care, duration of symptoms, diagnosis, and reason for seeking care in the PED. RESULTS: Two hundred patients were enrolled. Median age was 17 years (range, 1-22 years). Forty-five percent were African American. Forty-nine percent had Medicaid. Fifty percent identified a regular dentist, whereas 71% had a primary care physician. Thirty-four percent of patients 4 years and older had not seen a dentist in more than a year. Children younger than 13 years were more likely than teenagers to identify a regular dentist (odds ratio [OR] = 2.8; 95% confidence interval [CI], 1.3-6.1). Those with a regular medical provider were more likely to have a regular dentist (OR = 7.7; 95% CI, 3.4-18). The most common reasons for not going to a dentist were as follows: dentist closed, 34%; lack of dental insurance or money, 17%; and lack of a dentist, 16%. Patients with symptoms for more than 72 hours were more likely to cite lack of a dentist as their reason for coming to the PED (OR = 7.4; 95% CI, 1.9-33). CONCLUSIONS: Many pediatric patients do not have regular dental care, and this is associated with a lack of primary medical care. Access barriers to acute dental care include lack of insurance or funds, lack of a dentist, and limited hours of dental care sites. Improved insurance reimbursement, active enrollment of adolescents into preventive dental care, and expansion of provider hours may limit PED dental visits and improve the health of patients. PMID- 11386962 TI - Adolescent occupational toxic exposures: a national study. AB - BACKGROUND: While many previous studies describe workplace-associated injuries in adolescents, few focus on toxic exposures. Such incidents are unlikely to be reported to either federal or state agencies. However, poison control centers often get called about these poisonings and might serve as a resource for monitoring their occurrence. OBJECTIVE: To describe the frequency and severity of job-related toxic exposures involving adolescents, the specific toxic agents involved, and trends over time. METHODS: Occupational toxic exposures occurring in the United States between 1993 and 1997 were analyzed using the Toxic Exposure Surveillance System database compiled by the American Association of Poison Control Centers. Contingency tables with the chi(2) statistic were used to test bivariate associations. Logistic regression was performed to investigate trends over time. RESULTS: Of 301 228 workplace toxic exposures reported over 5 years, 8779 (3%) involved adolescents younger than 18 years. The most common agents involved were alkaline corrosives (13.2%), gases and fumes (12.0%), cleaning agents (9.7%), bleaches (8.3%), drugs (7.4%), acids (7.2%), and hydrocarbons (6.9%). The injuries were rated as severe in 14.2% of exposures, life-threatening in 0.3%, and there were 2 deaths. The proportionate frequency of occupational exposures occurring among adolescents vs adults increased over time (odds ratio, 1.003; P<.001). CONCLUSIONS: Adolescent occupational toxic exposures are an underrecognized hazard in the United States. Poison control center experience can be used to fill a gap in the surveillance of such injuries. PMID- 11386963 TI - Environmental influences, physical activity, and weight status in 8- to 16-year olds. AB - OBJECTIVE: To assess the association between vigorous physical activity, participation on sport teams and in exercise programs, television watching, family environment, and weight status in youth. DESIGN: Cross-sectional data from the Third National Health and Nutrition Examination Survey. PARTICIPANTS: A group of 2791 youth between the ages of 8 and 16 years who were enrolled in the Third National Health and Nutrition Examination Survey. MAIN OUTCOME MEASURES: Overweight was defined using age- and sex-specific 85th percentile of body mass index using Center for Disease Control and Prevention growth charts. RESULTS: Both males and females who had an overweight mother and or father were more likely to be overweight compared with youth who did not have an overweight parent. Females who watched 4 or more hours of television were more likely to be overweight than those who watched less than 4 hours. Males and 14- to 16-year-old females who participated in sport team and exercise programs were less likely to be overweight than their counterparts who did not participate. Also, females with larger families and males from families with higher family incomes were less likely to be overweight. CONCLUSIONS: These results suggest that family environment is associated with overweight in youth and that sport and exercise program participants are less like to be overweight and that, for females, increased television watching is related to overweight. PMID- 11386964 TI - Urinary thiobarbituric acid-reacting substances as potential biomarkers of intrauterine hypoxia. AB - BACKGROUND: Currently available clinical tools cannot accurately identify the extent of perinatal hypoxic injuries. During hypoxia, reactive oxygen species cause lipid peroxidation of cell membranes, yielding oxidation products that constitute thiobarbituric acid-reacting substances (TBARS). OBJECTIVE: To see if the concentrations of TBARS excreted in urine would be elevated during the first day of life in term and preterm infants following chronic hypoxia or acute asphyxia. DESIGN: Thiobarbituric acid-reacting substances levels were measured by a spectrophotometric assay in urine samples collected from term and near-term (>/= 34 weeks gestation, n = 22), and preterm (<34 weeks gestation, n = 52) infants on the first day of life. PATIENTS: Infants were admitted to the St Peter's University Hospital (New Brunswick, NJ) neonatal intensive care unit from July 1997 to January 1999. Acute asphyxia was defined as umbilical cord blood pH values less than 7.05, or Apgar scores of less than 5 at 5 minutes. Chronic hypoxia was defined as intrauterine growth retardation or low birth weight (small for gestational age) associated with pregnancy-induced hypertension or reversal of umbilical arterial blood flow. RESULTS: Among term infants, urinary TBARS levels were significantly increased following acute asphyxia (P =.02). Levels of TBARS also tended to be elevated following chronic hypoxia. Urinary TBARS levels in term infants tended to be increased in those requiring mechanical ventilation (P =.05) or delivery room resuscitation (P =.15), as well as in those passing intrauterine meconium (P =.13) or having clinical evidence of hypoxic-ischemic encephalopathy (P =.24). CONCLUSIONS: The results show a correlation between elevated urinary TBARS levels in term and near-term infants, and perinatal hypoxia (as determined by low Apgar scores or umbilical cord blood acidosis). We speculate that TBARS concentrations may be useful as a biomarker for perinatal hypoxic injury in newborns. Further studies are needed to determine whether elevations in TBARS levels are better predictors of the extent of hypoxic injury than existing markers. PMID- 11386966 TI - Special feature: picture of the month. PMID- 11386965 TI - Effects of phenobarbital on cerebral blood flow velocity after endotracheal suctioning in premature neonates. AB - OBJECTIVE: To examine the effect of phenobarbital administration on anterior cerebral artery blood flow velocity before and after endotracheal suctioning in premature neonates. DESIGN: Transcutaneous PO(2) (TcPO(2)), heart rate, mean arterial blood pressure (MABP), and Doppler velocimeter blood flow of the left anterior cerebral artery were measured before and immediately after 3 consecutive endotracheal suctioning procedures in premature neonates. Intravenous phenobarbital (20 mg/kg) was administered immediately after the first procedure. SETTING: Neonatal intensive care unit. PATIENTS: Nine neonates with a mean birth weight of 807 g (range, 620-1060 g) and a mean gestational age of 27 weeks (range, 25-30 weeks) were studied at age 8 to 12 hours. RESULTS: Transcutaneous PO(2) decreased in response to endotracheal suctioning at each of the suctioning procedures before and after phenobarbital was given (P<.001). Changes in heart rate were not observed. There were increases in MABP and area under the velocity curve (AUVC) per minute in response to endotracheal suctioning before but not after phenobarbital administration (P=.046). Use of phenobarbital lowered the overall peak systolic blood flow velocity in response to endotracheal suctioning (P =.02, analysis of variance, interactions for the effect of phenobarbital therapy on the response to suctioning). Changes in end-diastolic blood flow velocity were not observed. There were decreases in the differences before and after endotracheal suctioning for MABP at 2 and 4 hours and for AUVC and peak systolic blood flow velocity 4 hours after phenobarbital was given (P =.04). CONCLUSIONS: In very low-birth-weight neonates, endotracheal suctioning is associated with decreases in TcPO(2) and increases in MABP and AUVC. Treatment with phenobarbital attenuates the increases in MABP and AUVC but not the decreases in TcPO(2) after endotracheal suctioning. PMID- 11386967 TI - Special feature: radiological case of the month. PMID- 11386968 TI - Special feature: radiological case of the month. PMID- 11386969 TI - Special feature: pathological case of the month. PMID- 11386970 TI - Special feature: pathological case of the month. PMID- 11386976 TI - Incidence of unsuspected meningococcal disease. PMID- 11386980 TI - Efficacy of estradiol for the treatment of depressive disorders in perimenopausal women: a double-blind, randomized, placebo-controlled trial. AB - BACKGROUND: Results of previous studies suggest that estrogen improves somatic and mild depressive symptoms experienced by perimenopausal women. This study investigated the efficacy of 17beta-estradiol for the treatment of clinically significant depressive disorders in endocrinologically confirmed perimenopausal women. METHODS: Perimenopausal women (aged 40-55 years, with irregular menstrual periods and serum concentrations of follicle-stimulating hormone >25 IU/L), meeting criteria for major depressive disorder, dysthymic disorder, or minor depressive disorder, according to DSM-IV, were randomized to receive transdermal patches of 17beta-estradiol (100 microgram) or placebo in a 12-week, double blind, placebo-controlled study. A 4-week washout period followed the 12-week treatment phase. Outcome measures were the Montgomery-Asberg Depression Rating Scale and Blatt-Kupperman Menopausal Index scores. RESULTS: Fifty women were enrolled in the study; 26 met DSM-IV criteria for major depressive disorder, 11 for dysthymic disorder, and 13 for minor depressive disorder. Remission of depression was observed in 17 (68%) women treated with 17beta-estradiol compared with 5 (20%) in the placebo group (P =.001). Subjects responded similarly to estradiol treatment, regardless of DSM-IV diagnosis. Patients treated with estradiol sustained antidepressant benefit of treatment after the 4-week washout period, although somatic complaints increased in frequency and intensity. Treatment was well tolerated and adverse events were rare in both groups. CONCLUSION: Transdermal estradiol replacement is an effective treatment of depression for perimenopausal women. PMID- 11386981 TI - Sex and psychopharmacology: is natural estrogen a psychotropic drug in women? PMID- 11386982 TI - The role of serotonin transporter protein gene in antidepressant-induced mania in bipolar disorder: preliminary findings. AB - BACKGROUND: The occurrence of mania during antidepressant treatment is a key issue in the clinical management of bipolar disorder (BP). The serotonin transporter (5-HTT) is the selective site of action of most proserotonergic compounds used to treat bipolar depression. The 5-HTT gene (SLC6A4) has 2 known polymorphisms. The aim of this study was to investigate the role of the SLC6A4 variants in the pathogenesis of antidepressant-induced mania in BP. METHODS: Twenty-seven patients with a DSM-IV diagnosis of BP I or II, with at least 1 manic or hypomanic episode induced by treatment with proserotonergic antidepressants (IM+ group), were compared with 29 unrelated, matched patients with a diagnosis of BP I or II, who had been exposed to proserotonergic antidepressants without development of manic or hypomanic symptoms (IM- group). The 2 known polymorphisms of the SLC6A4 were genotyped, and allelic and genotypic association analyses were performed. RESULTS: With respect to the polymorphism in the promoter region (5HTTLPR), IM+ patients had an excess of the short allele (n = 34 [63%]) compared with IM- patients (n = 17 [29%]) (chi(2)(1), 12.77; P <.001). The genotypic association analysis showed a higher rate of homozygosity for the short variant in the IM+ group (n = 10 [37%]) than in the IM- group (n = 2 [7%]) and a lower rate of homozygosity for the long variant in the IM+ group (n = 3 [11%]) compared with the IM- group (n = 14 [48%]) (chi(2)(2), 12.43; P =.002). No associations were found for the polymorphism involving a variable number of tandem repeats. CONCLUSION: If these results are replicated, the 5HTTLPR polymorphism may become an important predictor of abnormal response to medication in patients with BP. PMID- 11386983 TI - Reduced glial cell density and neuronal size in the anterior cingulate cortex in major depressive disorder. AB - BACKGROUND: Glial cells are more numerous than neurons in the cortex and are crucial to neuronal function. There is evidence for reduced neuronal size in schizophrenia, with suggestive evidence for reduced glial cell density in mood disorders. In this investigation, we have simultaneously assessed glial cell density and neuronal density and size in the anterior cingulate cortex in schizophrenia, major depressive disorder, and bipolar disorder. METHODS: We examined tissue from area 24b of the supracallosal anterior cingulate cortex in 60 postmortem brain specimens from 4 groups of 15 subjects, as follows: major depressive disorder, schizophrenia, bipolar disorder, and normal controls. Glial cell density and neuronal size and density were examined in all subjects using the nucleator and the optical disector. RESULTS: Glial cell density (22%) (P =.004) and neuronal size (23%) (P =.01) were reduced in layer 6 in major depressive disorder compared with controls. There was some evidence for reduced glial density in layer 6 (20%) (P =.02) in schizophrenia compared with controls, before adjusting for multiple layerwise comparisons, but there were no significant changes in neuronal size. There was no evidence for differences in glial density or neuronal size in bipolar disorder compared with controls. Neuronal density was similar in all groups to that found in controls. CONCLUSION: These findings suggest that there is reduced frontal cortical glial cell density and neuronal size in major depressive disorder. PMID- 11386984 TI - Reductions in occipital cortex GABA levels in panic disorder detected with 1h magnetic resonance spectroscopy. AB - BACKGROUND: There is preclinical evidence and indirect clinical evidence implicating gamma-aminobutyric acid (GABA) in the pathophysiology and treatment of human panic disorder. Specifically, deficits in GABA neuronal function have been associated with anxiogenesis, whereas enhancement of GABA function tends to be anxiolytic. Although reported peripheral GABA levels (eg, in cerebrospinal fluid and plasma) have been within reference limits in panic disorder, thus far there has been no direct assessment of brain GABA levels in this disorder. The purpose of the present work was to determine whether cortical GABA levels are abnormally low in patients with panic disorder. METHODS: Total occipital cortical GABA levels (GABA plus homocarnosine) were assessed in 14 unmedicated patients with panic disorder who did not have major depression and 14 retrospectively age- and sex-matched control subjects using spatially localized (1)H-magnetic resonance spectroscopy. All patients met DSM-IV criteria for a principal current diagnosis of panic disorder with or without agoraphobia. RESULTS: Patients with panic disorder had a 22% reduction in total occipital cortex GABA concentration (GABA plus homocarnosine) compared with controls. This finding was present in 12 of 14 patient-control pairs and was not solely accounted for by medication history. There were no significant correlations between occipital cortex GABA levels and measures of illness or state anxiety. CONCLUSIONS: Panic disorder is associated with reductions in total occipital cortex GABA levels. This abnormality might contribute to the pathophysiology of panic disorder. PMID- 11386985 TI - Quality of medical care and excess mortality in older patients with mental disorders. AB - BACKGROUND: This study investigated whether differences in quality of medical care might explain a portion of the excess mortality associated with mental disorders in the year after myocardial infarction. METHODS: This study examined a national cohort of 88 241 Medicare patients 65 years and older who were hospitalized for clinically confirmed acute myocardial infarction. Proportional hazard models compared the association between mental disorders and mortality before and after adjusting 5 established quality indicators: reperfusion, aspirin, beta-blockers, angiotensin-converting enzyme inhibitors, and smoking cessation counseling. All models adjusted for eligibility for each procedure, demographic characteristics, cardiac risk factors and history, admission characteristics, left ventricular function, hospital characteristics, and regional factors. RESULTS: After adjusting for the potential confounding factors, presence of any mental disorder was associated with a 19% increase in 1-year risk of mortality (hazard ratios [HR], 1.19; 95% confidence interval [CI], 1.04-1.36). After adding the 5 quality measures to the model, the association was no longer significant (HR, 1.10; 95% CI, 0.96-1.26). Similarly, while schizophrenia (HR, 1.34; 95% CI, 1.01-1.67) and major affective disorders (HR, 1.11; 95% CI, 1.02 1.20) were each initially associated with increased mortality, after adding the quality variables, neither schizophrenia (HR, 1.23; 95% CI, 0.86-1.60) nor major affective disorder (HR, 1.05; 95% CI, 0.87-1.23) remained a significant predictor. CONCLUSIONS: Deficits in quality of medical care seemed to explain a substantial portion of the excess mortality experienced by patients with mental disorders after myocardial infarction. The study suggests the potential importance of improving these patients' medical care as a step toward reducing their excess mortality. PMID- 11386986 TI - Incidence of cancer among persons with schizophrenia and their relatives. AB - BACKGROUND: It has repeatedly been reported that the risk for cancer in patients with schizophrenia is different from that of the general population, specifically a lower risk for lung cancer despite increased smoking. Confirmation of these associations could lead to hypotheses on shared risk or protective factors, either genetic or environmental. METHODS: From Finland's National Hospital Discharge and Disability Pension registers, Helsinki, we identified a cohort of 26 996 individuals born between 1940 and 1969 and treated for schizophrenia between 1969 and 1991. They were followed up for cancer from 1971 to 1996 by record linkage with the Finnish Cancer Registry, yielding 446 653 person-years at risk, and standardized incidence ratios (SIRs) were calculated. Likewise, 39 131 parents and 52 976 siblings of the patients with schizophrenia were followed up to explore familial genetic hypotheses on deviations in cancer risk. RESULTS: In patients with schizophrenia, an increased overall cancer risk was found (724 cases observed vs 619 expected; SIR, 1.17; 95% confidence interval [CI], 1.09 1.25). Half of the excess cases were attributable to lung cancer (SIR, 2.17; 95% CI, 1.78-2.60), and the strongest relative increase in risk was in pharyngeal cancer (SIR, 2.60; 95% CI, 1.25-4.77). Cancer incidence in siblings (SIR, 0.89; 95% CI, 0.83-0.94) and parents (SIR, 0.91; 95% CI, 0.89-0.93) was consistently lower than that in the general population. CONCLUSION: Although specific lifestyle factors, particularly tobacco smoking and alcohol consumption, probably account for the increased cancer risk in patients with schizophrenia, the decreased risk in relatives would be compatible with a postulated genetic risk factor for schizophrenia offering selective advantage to unaffected relatives. PMID- 11386987 TI - Schizophrenia and cancer: is there a need to invoke a protective gene? PMID- 11386988 TI - Schizophrenia and schizophrenia-spectrum personality disorders in the first degree relatives of children with schizophrenia: the UCLA family study. AB - BACKGROUND: This study tested the hypothesis that childhood-onset schizophrenia (COS) is a variant of adult-onset schizophrenia (AOS) by determining if first degree relatives of COS probands have an increased risk for schizophrenia and schizotypal and paranoid personality disorders. METHODS: Relatives of COS probands (n = 148) were compared with relatives of attention deficit/hyperactivity disorder (ADHD) (n = 368) and community control (n = 206) probands. Age-appropriate structured diagnostic interviews were used to assign DSM-III-R diagnoses to probands and their relatives. Family psychiatric history was elicited from multiple informants. Diagnoses of relatives were made blind to information about probands' diagnoses. Final consensus diagnoses, which integrated family history, direct interview information, and medical records, are reported in this article. RESULTS: There was an increased lifetime morbid risk for schizophrenia (4.95% +/- 2.16%) and schizotypal personality disorder (4.20% +/- 2.06%) in the parents of COS probands compared with parents of ADHD (0.45% +/ 0.45%, 0.91% +/- 0.63%) and community control (0%) probands. The parents of COS probands diagnosed as having schizophrenia had an early age of first onset of schizophrenia. Risk for avoidant personality disorder (9.41% +/- 3.17%) was increased in the parents of COS probands compared with parents of community controls (1.67% +/- 1.17%). CONCLUSIONS: The psychiatric disorders that do and do not aggregate in the parents of COS probands are remarkably similar to the disorders that do and do not aggregate in the parents of adults with schizophrenia in modern family studies. These findings provide compelling support for the hypothesis of etiological continuity between COS and AOS. PMID- 11386989 TI - The prevalence of personality disorders in a community sample. AB - BACKGROUND: To our knowledge, no previous studies of personality disorders (PDs) in a large representative sample of the common population have been conducted. METHODS: A representative sample of 2053 individuals between the ages of 18 and 65 years in Oslo, the capital of Norway, was studied from 1994 to 1997. Information about PDs was obtained by means of the Structured Interview for DSM III-R Personality Disorders, in conjunction with an interview recording demographic data. The subjects were interviewed primarily at home, but in some instances, also at the clinic. RESULTS: The prevalence of PDs was 13.4% (SE, 0.7). The prevalence rates (SEs) for specific PDs, irrespective of whether a person had 1 or more PD, were: paranoid, 2.4% (0.3); schizoid, 1.7% (1.6); schizotypal, 0.6% (0.2); antisocial, 0.7% (0.2); sadistic, 0.2% (0.1); borderline, 0.7% (0.2); histrionic, 2.0% (0.3); narcissistic, 0.8; (0.2); avoidant, 5.0% (0.5); dependent, 1.5% (0.3); obsessive-compulsive: 2.0% (0.3); passive-aggressive, 1.7% (0.3); self-defeating, 0.8%, (0.2). The prevalence of PDs was highest among subjects with only a high school education or less, and living without a partner in the center of the city. CONCLUSIONS: Personality disorders were found to be prevalent, with avoidant, schizoid, and paranoid PDs more common, and borderline PD less common than what is usually reported. Personality disorders tend to be more frequent among single individuals from the lower socioeconomic classes in the center of the city. It is impossible to determine what is cause and what is consequence from a cross-sectional study. PMID- 11386990 TI - The structure and stability of common mental disorders: the NEMESIS study. AB - BACKGROUND: We analyzed the underlying latent structure of 12-month DSM-III-R diagnoses of 9 common disorders for the general population in the Netherlands. In addition, we sought to establish (1) the stability of the latent structure underlying mental disorders across a 1-year period (structural stability) and (2) the stability of individual differences in mental disorders at the level of the latent dimensions (differential stability). METHODS: Data were obtained from the first and second measurement of the Netherlands Mental Health Survey and Incidence Study (NEMESIS) (response rate at baseline: 69.7%, n = 7076; 1 year later, 79.4%, n = 5618). Nine common DSM-III-R diagnoses were assessed twice with the Composite International Diagnostic Interview with a time lapse of 1 year. Using structural equation modeling, the number of latent dimensions underlying these diagnoses was determined, and the structural and differential stability were assessed. RESULTS: A 3-dimensional model was established as having the best fit: a first dimension underlying substance use disorders (alcohol dependence, drug dependence); a second dimension for mood disorders (major depression, dysthymia), including generalized anxiety disorder; and a third dimension underlying anxiety disorders (simple phobia, social phobia, agoraphobia, and panic disorder). The structural stability of this model during a 1-year period was substantial, and the differential stability of the 3 latent dimensions was considerable. CONCLUSIONS: Our results confirm the 3-dimensional model for 12 month prevalence of mental disorders. Results underline the argument for focusing on core psychopathological processes rather than on their manifestation as distinguished disorders in future population studies on common mental disorders. PMID- 11386991 TI - Use of suprathreshold electroconvulsive therapy. PMID- 11386992 TI - Electrode placement and electroconvulsive therapy: a search for the chimera. PMID- 11386995 TI - Frontal lobe pathology and antisocial personality disorder. PMID- 11386997 TI - Long-term management and outcome of parathyroidectomy for sporadic primary multiple-gland disease. AB - HYPOTHESIS: For a specific subset of patients with sporadic primary multiple gland parathyroid disease, subtotal parathyroidectomy results in long-term normocalcemia in the majority of patients, with a minimal complication rate. DESIGN: Retrospective analysis of outcomes in patients undergoing parathyroidectomy performed by a single surgeon (A.E.G.) between 1984 and 1999. SETTING: A multidisciplinary endocrine service based at a tertiary referral center. PATIENTS: Patients undergoing subtotal parathyroidectomy for primary hyperparathyroidism due to sporadic multiple-gland disease identified from a single surgeon's operative records (A.E.G.). MAIN OUTCOME MEASURES: Data analyzed included demographic factors, operative and pathologic findings, and postoperative and long-term clinical and laboratory results, including calcium and intact parathyroid hormone levels. RESULTS: Of 379 patients undergoing parathyroidectomy for hyperparathyroidism between 1984 and 1999, 49 (13%) had sporadic multiple-gland disease. Median preoperative calcium and intact parathyroid hormone (iPTH) levels were 2.7 mmol/L (10.8 mg/dL) and 11.79 pmol/L, respectively. Postoperative calcium and iPTH levels were available in 39 patients, and median values were 2.28 mmol/L (9.1 mg/dL) and 2.84 pmol/L, respectively. Long-term follow-up was available for 36 patients (73%), and duration ranged from 6 to 180 months (median, 44 months). Median calcium and iPTH levels at follow-up were 2.3 mmol/L (9.2 mg/dL) and 3.26 pmol/L, respectively, with 3 (8%) of 36 patients having evidence of persistent or recurrent hyperparathyroidism. No patient had biochemical evidence of hypoparathyroidism at long-term follow-up. Five patients (14%) had persistent elevated iPTH levels (range, 8.11-10.95 pmol/L) and normal calcium levels. CONCLUSIONS: Subtotal parathyroidectomy for sporadic primary multiple-gland disease resulted in a long term normocalcemia rate of 92%, with minimal complications. Selective subtotal parathyroidectomy can yield excellent long-term results in patients with multiple gland disease. PMID- 11386998 TI - Lessons learned in adopting endovascular techniques for treating abdominal aortic aneurysm. AB - HYPOTHESIS: Endovascular exclusion of abdominal aortic and common iliac aneurysms can be performed safely, and in the short term represents a feasible alternative to traditional, open aneurysm repair. PATIENTS AND METHODS: Forty-one patients were treated with endovascular grafts for 39 abdominal aortic and 2 common iliac artery aneurysms. RESULTS: All devices were successfully deployed. The size of the abdominal aortic aneurysms varied from 4.9 to 11.9 cm (average, 6.13 cm). The median procedure time was 195 minutes. There was one iliac artery rupture, which required celiotomy for repair. The hospital stay varied from 2 to 39 days (average, 6.7 days). The perioperative mortality rate was 2.4%. Sixteen patients (39%) had groin wound complications. Ten patients (24%) had evidence of contrast (endoleak) within the aneurysm sac on completion of the procedure. There were no obvious direct leaks from either the point of proximal or distal fixation. Seven of these endoleaks have resolved spontaneously. Two patients required additional procedures in the postoperative period to treat endoleak. The final patient has evidence of persistent endoleak on 3-month surveillance computed tomography scan. Major late problems occurred in 3 patients. CONCLUSION: Patients with large abdominal aortic aneurysms and considerable cardiac comorbidity can safely undergo endovascular aneurysm repair. Femoral groin wound complications resulting in prolonged hospitalization remain the major cause of perioperative morbidity. In contradistinction to open aneurysm repair, long-term surveillance is essential to detect migration of the device and identify flow within the residual aneurysm sac-complications that could lead to aneurysm rupture following endovascular repair. PMID- 11386999 TI - Patency and limb salvage after infrainguinal bypass with severely compromised ("blind") outflow. AB - HYPOTHESIS: Infrainguinal graft patency and limb salvage are adversely affected by severely compromised outflow. DESIGN: Retrospective review of all infrainguinal bypass procedures performed at a single institution during a 5-year period. SETTING: University teaching hospital. PATIENTS: Two hundred seventy-four patients underwent infrainguinal bypass for limb salvage (351 grafts in 307 limbs). INTERVENTIONS: All infrainguinal bypasses originated from a femoral artery. The distal anastomosis in 279 grafts was located in an artery with at least 1 patent outflow vessel with anatomically normal end-artery runoff (Society for Vascular Surgery/International Society for Cardiovascular Surgery ad hoc committee runoff score, 1-9). The distal anastomosis of 72 grafts was located in an artery with only collateral outflow ("blind bypass"; runoff score, 10). MAIN OUTCOME MEASURES: Perioperative morbidity and mortality, primary-assisted and secondary graft patency, limb salvage, and survival. RESULTS: All data are presented as mean +/- SEM. Patients undergoing blind bypass were older (age, 70 +/- 2 vs. 66 +/- 1 years; P <.05) and had a higher incidence of hypertension (90% vs 70%; P <.05) and end-stage renal disease (24% vs. 13%; P <.05). Comparing patients undergoing blind bypass to bypass with at least 1 patent outflow vessel, there were no differences in the use of nonautogenous conduits (50% vs 59%; P =.21) or postoperative warfarin (30% vs 32%; P =.69), or in perioperative mortality rates (2.7% vs 3.2%; P =.79). After a median follow-up of 13 months (range, 0-60 months), 2-year secondary graft patency for the entire group was 63% +/- 4%. The secondary patency rate of blind bypass grafts was no different from that of grafts with at least 1 patent outflow vessel (67% +/- 7% vs. 64% +/- 4%; P was not significant). However, the 2-year limb salvage rate in limbs with blind outflow was significantly worse than in limbs with at least 1 patent outflow vessel (67% +/- 7% vs. 76% +/- 3%; P =.04). CONCLUSION: Acceptable long-term patency rates can be achieved in infrainguinal bypass grafts with blind outflow, although blind outflow remains a marker for subsequent limb loss in the chronically ischemic leg. PMID- 11387000 TI - Postobstructive chronic pancreatitis: results with distal resection. AB - HYPOTHESIS: For most patients with chronic obstructive pancreatitis, distal pancreatectomy confers pain relief. DESIGN: Retrospective case series. Follow-up was complete in 80% of study subjects (mean follow-up, 6.7 years). SETTING: Tertiary care center. PATIENTS: Among 484 patients with chronic pancreatitis undergoing operation from 1976 to 1997, 40 with postobstructive chronic pancreatitis were identified. Criteria for selection included an isolated, dominant major pancreatic duct stricture or cutoff, changes of chronic pancreatitis in the distal pancreas, and ostensibly normal parenchyma without calcification in the proximal gland. The patients were reviewed with regard to operative procedure, postoperative course, and outcome. MAIN OUTCOME MEASURES: Outcome measures included degree of pain relief, morbidity and mortality of operation, survival, rates of endocrine and exocrine insufficiency, and ability to return to work and/or normal activities. RESULTS: All but 1 of the 40 patients had abdominal pain, and 20 (50%) had recurrent episodes of acute pancreatitis. Suspicion of malignancy was a concern in 16 patients (40%). Thirty-eight patients underwent distal pancreatectomy; 1 had a central resection and another a Roux-en Y cystojejunostomy. There was no operative mortality, but significant morbidity occurred in 15%. Among 31 patients with preoperative pain in whom long-term follow-up was available, complete or significant pain relief was achieved in 25 (81%); 74% returned to normal social function, but about half had some element of pancreatic insufficiency. CONCLUSIONS: Distal pancreatectomy is a safe procedure and achieves pain relief and good quality of life in a large percentage of patients (80%) with presumed postobstructive chronic pancreatitis. However, some of these patients with chronic pancreatitis involving the entire gland have disease masquerading as postobstructive chronic pancreatitis secondary to an ostensibly isolated dominant pancreatic ductal stricture. PMID- 11387001 TI - Childbearing and child care in surgery. AB - HYPOTHESIS: The responsibility for childbearing and child care has a major effect on general surgical residency and subsequent surgical practice. METHODS: A survey of all graduates from a university general surgical training program between 1989 and 2000. RESULTS: Twenty-seven women and 44 men completed general surgical training at our university during the period, and 42 (59%) responded to our survey. The age at completion of the residency was 34.0 +/- 2.2 years for men and 33.9 +/- 2.8 years for women. During residency, 64% (14/22) of the men and 15% (3/20) of the women had children. At the time of the survey, 21 (95%) of the men and 8 (40%) of the women had children. Most residents (24 [57%] of 42) relied on their spouse for child care. During surgical practice, 18 (43%) indicated that they rely on their spouse; 19 (45%) use day care, home care, or both; and (8%) of 26 are unsatisfied with their current child care arrangement. During training, 38% (5/13) of men and 67% (2/3) of women took time off for maternity leave, paternity leave, or child care. Two of 3 surgeons would like to have had more time off during residency; most men (70%, or 7 of 10) recommended a leave of 1 to 3 months, and all women preferred a 3-month maternity or child care leave of absence. During surgical practice, only 12% (2/17) of men but 64% (7/11) of women have taken time off for either childbearing or child care. Half of the respondents (21/42) have a formal leave of absence policy at work, 52% (11/21) of which are paid leave programs. Although the workweek of our practicing graduates is 69 +/- 16 hours for men and 64 +/- 12 hours for women, 62% (26/42) spend more than 20 hours per week parenting. More than 80% (27/32) would consider a part time surgical practice for more parenting involvement; one third of the responders suggested that 30 hours a week constitutes a reasonable part-time practice, one third preferred fewer than 30 hours, and one third favored more than 30 hours per week. Data are presented as mean +/- SD. CONCLUSIONS: Childbearing and child care may have an enormous impact on one's decision to pursue a career in surgery. To attract and retain the best candidates for future surgeons, formal policies on the availability of child care services in the residency program and the workplace should be studied and implemented. Furthermore, national studies are needed to define appropriate, acceptable workweeks for part-time or flexible practices and the duration of leaves of absence for childbearing or child care. PMID- 11387003 TI - Adult-onset nesidioblastosis causing hypoglycemia: an important clinical entity and continuing treatment dilemma. AB - HYPOTHESIS: Nesidioblastosis is an important cause of adult hyperinsulinemic hypoglycemia, and control of this disorder can often be obtained with a 70% distal pancreatectomy. DESIGN: The records of all adult patients operated on for hypoglycemia between 1974 and 1999 were reviewed retrospectively. Patients with the pathologic diagnosis of nesidioblastosis were contacted for follow-up (1.5-21 years) and are presented. Patients' results were compared with those of 36 other individuals with this disorder who were previously reported in the literature. SETTING: The University of Chicago Medical Center (Chicago, Ill), a tertiary care facility. PATIENTS: A consecutive sample of all patients operated on for hypoglycemia. INTERVENTIONS: Seventy percent distal pancreatectomy for all patients with nesidioblastosis, and maintenance therapy with verapamil hydrochloride for 2 patients. MAIN OUTCOME MEASURES: Achievement of normoglycemia with and without medication, development of insulin-dependent diabetes mellitus, pancreatic exocrine insufficiency, and need for reoperation. RESULTS: Of 32 adult patients who underwent surgical exploration for hyperinsulinemic hypoglycemia at our institution, 27 (84%) were found to have 1 or more insulinomas, and 5 (16%) were diagnosed with nesidioblastosis. Each patient with nesidioblastosis underwent a 70% distal pancreatectomy. Follow-up duration for the 5 patients ranged from 1.5 to 21 years, with 3 patients (60%) asymptomatic and taking no medications, and 2 patients (40%) experiencing some recurrences of hypoglycemia. The 2 patients with recurrences are now successfully treated with a calcium channel blocker, an approach, to our knowledge, never before reported for adult onset nesidioblastosis. CONCLUSIONS: Nesidioblastosis is an uncommon but clinically important cause of hypoglycemia in the adult population, and must always be considered in a patient with a presumptive preoperative diagnosis of insulinoma. This study indicates that a 70% distal pancreatectomy is often successful in controlling hypoglycemia, and rarely results in diabetes mellitus. However, the optimal treatment of this disorder remains to be determined. PMID- 11387004 TI - Long-term results of metallic stents for benign biliary strictures. AB - BACKGROUND: Historically, surgical correction has been the treatment of choice for benign biliary strictures (BBS). Self-expandable metallic stents (MSs) have been useful for inoperable malignant biliary strictures; however, their use for BBS is controversial and their natural history unknown. HYPOTHESIS: To test our hypothesis that MSs provide only short-term benefit, we examined the long-term outcome of MSs for the treatment of BBS. Our goal was to develop a rational approach for treating BBS. DATA EXTRACTION: Between July 1990 and December 1995, 15 patients had MSs placed for BBS and have been followed up for a mean of 86.3 months (range, 55-120 months). The mean age of the patients was 66.6 years and 12 were women. Stents were placed for surgical injury in 5 patients and underlying disease in 10 patients (lithiasis, 7; pancreatitis, 2; and primary sclerosing cholangitis, 1). One or more MSs (Gianturco-Rosch "Z" for 4 patients and Wallstents for 11 patients) were placed by percutaneous, endoscopic, or combined approaches. We considered patients to have a good clinical outcome if the stent remained patent, they required 2 or fewer invasive interventions, and they had no biliary dilation on subsequent imaging. DATA SYNTHESIS: Metallic stents were successfully placed in all 15 patients, and the mean patency rate was 30.6 months (range, 7-120 months). Five patients (33%) had a good clinical result with stent patency from 55 to 120 months. Ten patients (67%) required more than 2 radiologic and/or endoscopic procedures for recurrent cholangitis and/or obstruction (range, 7-120 months). Five of the 10 patients developed complete stent obstruction at 8, 9, 10, 15, and 120 months and underwent surgical removal of the stent and bilioenteric anastomosis. Four of these 5 patients had strictures from surgical injuries. The patient who had surgical removal 10 years after MS placement developed cholangiocarcinoma. CONCLUSIONS: Surgical repair remains the treatment of choice for BBS. Metallic stents should only be considered for poor surgical candidates, intrahepatic biliary strictures, or failed attempts at surgical repair. Most patients with MSs will develop recurrent cholangitis or stent obstruction and require intervention. Chronic inflammation and obstruction may predispose the patient to cholangiocarcinoma. PMID- 11387006 TI - Computed tomography and ultrasonography in the diagnosis of appendicitis: when are they indicated? AB - HYPOTHESIS: Relative merits and indications exist for ultrasonography (US) and computed tomography (CT) in the diagnosis of appendicitis. DESIGN: Prospective study. SETTING: General community and tertiary care hospital. METHODS: Ninety nine patients (50 males and 49 females) were studied. Following consent, the initial disposition was recorded, designating the patient for operation, observation, or discharge from the hospital. Each patient was studied by CT and US. Studies were independently evaluated by 2 radiologists, and the results were designated as positive, negative, or equivocal. The surgeon reevaluated patients before and after learning the results of US and CT, recording whether the CT scan, US, or reexamination influenced the final disposition. RESULTS: Fifty patients had appendicitis; 6 appendixes were perforated. The initial clinical impression called for 44 operations, 49 observations, and 6 discharges. Thirty four patients had their treatment plan changed from the initial disposition. Ultrasonography did not affect the initial impression. In contrast, 18 patients were rediagnosed solely on CT scan findings. Seven patients were rediagnosed by reexamination. Of 44 patients initially designated for operation, the CT scan and reexamination spared 6 females from surgery; the negative appendectomy rate potentially decreased from 50% to 17% (P =.03). The CT scan, US, or reexamination failed to spare 2 males from exploration with negative results. Of the 49 patients initially designated for observation, 23 were rediagnosed after reevaluation, 13 were discharged from the hospital, and 10 underwent expedient operation. One patient was spared from inappropriate discharge from the hospital. The reliability of the CT scan was good, with high sensitivities and specificities. Equivocal scan results lowered the diagnostic value. CONCLUSIONS: Selective use of a CT scan with a second examination can improve the diagnostic accuracy and management of suspected cases of appendicitis by (a) reducing the negative appendectomy rate in females, (b) moving patients from observation to earlier operation or discharge from the hospital, and (c) preventing inappropriate discharge of patients with appendicitis. PMID- 11387007 TI - Avoidance of abdominal compartment syndrome in damage-control laparotomy after trauma. AB - HYPOTHESIS: Abdominal compartment syndrome (ACS) is a morbid complication of damage-control laparotomy. Moreover, the technique of abdominal closure influences the frequency of ACS. DESIGN: Retrospective cohort study. SETTING: Urban level I trauma center. PATIENTS: We studied 52 patients with trauma who required damage-control laparotomy during the 5 years ending December 31, 1999, and who survived longer than 48 hours. MAIN OUTCOME MEASURES: Abdominal compartment syndrome, acute respiratory distress syndrome (ARDS), and multiple organ failure (MOF). RESULTS: Mean (+/- SD) age was 33 +/- 2 years; 38 (73%) were male. Mechanism of injury was blunt in 29 patients (56%), and mean (+/- SD) Injury Severity Score was 28 +/- 2. Development of ARDS and/or MOF was seen in 23 patients (44%); ARDS and MOF increased mortality from 12% (3/26) to 42% (11/26). Abdominal compartment syndrome was a common complication (17/52), and was associated with an increase in ARDS and/or MOF (12 patients [71%] vs 11 patients [31%] without ACS; P =.02, chi(2) test) and death (6 [35%] vs 8 patients [23%] without ACS). Primary fascial closure (n = 10) at the initial laparotomy was associated with ACS in 8 (80%) (P =.001, chi(2) test) and ARDS and/or MOF in 9 (90%) (P =.01, chi(2) test); skin closure (n = 25), with ACS in 6 (24%) and ARDS/MOF in 9 (36%); and Bogota bag closure (n = 17), with ACS in 3 (18%) and ARDS/MOF in 8 (47%). CONCLUSIONS: Damage-control laparotomy is associated with frequent complications. In particular, ACS is a serious complication that increases ARDS and/or MOF and mortality. Avoiding primary fascial closure at the initial laparotomy can minimize the risk for ACS. PMID- 11387009 TI - Spontaneous rupture of hepatocellular carcinoma and vascular injury. AB - HYPOTHESIS: Because spontaneous rupture of hepatocellular carcinoma (HCC) is one kind of bleeding complication related to the blood vessels, the possible mechanism of this rupture should occur on the blood vessel itself. Our hypothesis, which has not yet been investigated, is that the vascular integrity of HCC might be damaged during vascular injury. DESIGN: We examined semiquantitatively the expression of von Willebrand factor, elastin, neutrophil elastase, type IV collagen, and collagenase in 23 specimens of HCC with spontaneous rupture by immunohistochemistry, and compared them with 30 specimens of HCC without rupture. RESULTS: There was a significant decrease of von Willebrand factor, proliferation of degenerated elastin, abnormal distribution of neutrophil elastase, degradation of type IV collagen, and increase in collagenase production around the blood vessels in ruptured HCC. Since the decreased expression of von Willebrand factor is an indicator of vascular injury and elastase and collagenase are present in inflammatory processes, we postulate that the vascular injury probably exists before spontaneous rupture of HCC occurs. The blood vessel dysfunction resulting from the degeneration of elastin and the degradation of type IV collagen can render the blood vessels stiff and weak, causing them to split easily when the vascular load increases from hypertension or minor mechanical trauma. CONCLUSION: Spontaneous rupture of HCC may be related to the vascular dysfunction. PMID- 11387010 TI - Characteristics of the sentinel lymph node in breast cancer predict further involvement of higher-echelon nodes in the axilla: a study to evaluate the need for complete axillary lymph node dissection. AB - BACKGROUND: Sentinel lymph node (SLN) biopsy techniques provide accurate nodal staging for breast cancer. In the past, complete lymph node dissection (CLND) (levels 1 and 2) was performed for breast cancer staging, although the therapeutic benefit of this more extensive procedure has remained controversial. HYPOTHESIS: It has been demonstrated that if the axillary SLN has no evidence of micrometastases, the nonsentinel lymph nodes (NSLNs) are unlikely to have metastases. OBJECTIVE: To determine which variables predict the probability of NSLN involvement in patients with primary breast carcinoma and SLN metastases. METHODS: An analysis of 101 women with SLN metastases and subsequent CLND was performed. Variables included size of the primary tumor, tumor volume in the SLN, staining techniques used to initially identify the micrometastases (cytokeratin immunohistochemical vs hematoxylin-eosin), number of SLNs harvested, and number of NSLNs involved with the metastases. Tumor size was determined by the invasive component of the primary tumor. Patients with ductal carcinoma in situ who were upstaged with cytokeratin staining were considered to have stage T1a tumors. RESULTS: Sentinel lymph node micrometastases (<2 mm) detected initially by cytokeratin staining were associated with a 7.6% (2/26) incidence of positive CLND compared with a 25% (5/20) incidence when micrometastases were detected initially by routine hematoxylin-eosin staining. Sentinel lymph node micrometastases, regardless of identification technique, inferred a risk of 15.2% (7/46) for NSLN involvement. As the volume of tumor in the SLN increased (ie, <2 mm, >2 mm, grossly visible tumor), so did the risk of NSLN metastases (P<.001). CONCLUSIONS: Our study demonstrated that patients with micrometastases detected initially by cytokeratin staining had low-volume disease in the SLN with a small chance of having metastases in higher-echelon nodes in the regional basin other than the SLN. Characteristics of the SLN can provide information to determine the need for a complete axillary CLND. Complete lymph node dissection may not be necessary in patients with micrometastases detected initially by cytokeratin staining since the disease is confined to the SLN 92.4% of the time. However, the therapeutic value of CLND in breast cancer remains to be determined by further investigation. PMID- 11387012 TI - A prospective longitudinal study of quality of life after resection of hepatocellular carcinoma. AB - HYPOTHESIS: Hepatic resection improves quality of life (QOL) in patients with resectable hepatocellular carcinoma (HCC). DESIGN: A prospective longitudinal study. SETTING: A university teaching hospital. PATIENTS: Sixty-six consecutive patients undergoing resection of HCC, and 10 patients with unresectable HCC found after surgical exploration who were subsequently treated with transarterial chemoembolization (control group). MAIN OUTCOME MEASURE: Serial measurements of preoperative and postoperative QOL using the Functional Assessment of Cancer Therapy-General (FACT-G) Questionnaire for up to 2 years after surgery (at 3, 6, 9, 12, 18, and 24 months). RESULTS: Among the 66 patients with resectable HCC, the mean postoperative QOL scores at 3 months after surgery were significantly higher than the mean preoperative QOL scores in domains related to physical, social, and emotional well-being and relationship with physicians. The mean total QOL score increased from 83.5 (SD, 9.4) before surgery to 94.1 (SD, 7.7) at 3 months after surgery (P <.001). No significant change of QOL scores at 3 months after surgery was observed in the control group. Twenty patients in the resected group died of early recurrence within 2 years after surgery, but their mean postoperative QOL scores remained higher than the preoperative QOL scores for most assessment times. In contrast, in the control group, the mean total QOL scores became significantly lower than the preoperative scores, starting 9 months after surgery. Forty-six patients in the resected group completed all QOL assessments. At all postoperative assessments, their mean QOL scores were higher than preoperative scores. Recurrence developed in 13 of the 46 patients within the 2-year study, and there was significant deterioration of QOL over time among these 13 (P <.001), whereas no significant change in QOL over time was observed among the 33 recurrence-free patients. Of various clinicopathologic factors, only advanced pTNM stage was significantly predictive of deterioration of QOL over time after resection of HCC. CONCLUSIONS: Hepatic resection results in significant enhancement of QOL in patients with HCC. Development of recurrence is the main factor leading to deterioration in QOL over time after resection of HCC. PMID- 11387013 TI - Laparoscopic hepatic artery infusion pump placement. AB - Patients with metastatic colorectal cancer limited to the liver are candidates for regional chemotherapy with implantable hepatic artery infusion (HAI) pumps. The poor prognosis of these patients, and the requirement of a laparotomy for placement, has deterred many oncologists from referral for HAI pump implantation. Minimally invasive surgical techniques are particularly well suited for the task of HAI pump placement in patients who may not tolerate the additional physiologic stress of a major surgical intervention. Advances in laparoscopic techniques allow pumps to be implanted safely and effectively, replicating the well described tenets of open pump placement. The principal steps of the operation include a thorough laparoscopic evaluation to exclude extrahepatic disease, complete vascular isolation of the hepatic and gastroduodenal arteries, ligation of aberrant hepatic vessels, secure cannulation of the gastroduodenal artery, and confirmation of complete hepatic perfusion without extrahepatic perfusion. We describe the procedure and briefly review our clinical experience. We believe that the benefits typically derived from minimally invasive approaches (less pain, fewer perioperative complications, shorter hospitalization, faster recovery, and potentially less immune suppression) will be seen in these patients as well. If so, a completely laparoscopic approach to regional treatment of the liver may extend survival and improve the quality of life of patients whose prognosis is poor regardless of treatment. Controlled trials will be required to evaluate the added value of a laparoscopic approach to the placement of the hepatic artery pump. PMID- 11387014 TI - Special feature: image of the month. Diagnosis: acute left-sided appendicitis. PMID- 11387015 TI - Surgical reminiscences: five short stories. PMID- 11387016 TI - Moments in surgical history: a shoulder amputation in 1813. PMID- 11387017 TI - Progress and animal welfare: conflicting goals or complementary opportunities? PMID- 11387018 TI - Cytotoxicity of the MEIC reference chemicals in antioxidant-enriched, rat hepatoma-derived Fa32 cells. AB - Since vitamin E increases the antioxidant status of cells, its influence on cytotoxicity was investigated. The neutral red uptake (NRU) inhibition effects of 39 MEIC reference chemicals were measured after treatment of rat hepatoma-derived Fa32 cells in the presence of vitamin E for 30 minutes. The results were quantified in terms of the NI50, the concentration of test compound required to reduce the NRU by 50%. Sodium chloride was the only chemical that was more toxic in the presence of vitamin E. This effect was related to the concentration of vitamin E in the cell culture medium. A vitamin E dose-related response was also observed for the decreased toxicity of paracetamol and caffeine. Glutathione levels were slightly increased in the presence of vitamin E, which could contribute to the protective effect of vitamin E. Of the remaining chemicals, 50% were less toxic in the presence of vitamin E, but the correlation with the acute human toxicity data of the MEIC study was not improved. The results imply that reactive oxygen species interfere with the toxicity of a high proportion of toxic chemicals. The assay described provides a quick and easy method for checking whether reactive oxygen species contribute to the toxicity of a chemical. PMID- 11387019 TI - The use of cultured hepatocytes to investigate the metabolism of drugs and mechanisms of drug hepatotoxicity. AB - Hepatotoxins can be classified as intrinsic when they exert their effects on all individuals in a dose-dependent manner, and as idiosyncratic when their effects are the consequence of an abnormal metabolism of the drug by susceptible individuals (metabolic idiosyncrasy) or of an immune-mediated injury to hepatocytes (allergic hepatitis). Some xenobiotics are electrophilic, and others are biotransformed by the liver into highly reactive metabolites that are usually more toxic than the parent compound. This activation process is the key to many hepatotoxic phenomena. Mitochondria are a frequent target of hepatotoxic drugs, and the alteration of their function has immediate effects on the energy balance of cells (depletion of ATP). Lipid peroxidation, oxidative stress, alteration of Ca(2+) homeostasis, and covalent binding to cell macromolecules are the molecular mechanisms that are frequently involved in the toxicity of xenobiotics. Against these potential hazards, cells have their own defence mechanisms (for example, glutathione, DNA repair, suicide inactivation). Ultimately, toxicity is the balance between bioactivation and detoxification, which determines whether a reactive metabolite elicits a toxic effect. The ultimate goal of in vitro experiments is to generate the type of scientific information needed to identify compounds that are potentially toxic to man. For this purpose, both the design of the experiments and the interpretation of the results are critical.] PMID- 11387020 TI - The DRAG test: an assay for detection of genotoxic damage. AB - A high throughput assay (the DRAG test) is described, which could be a useful tool for the detection of repairable DNA adducts, and which is based on the inhibition of the growth of DNA repair-deficient Chinese hamster ovary (CHO) cells. The cytotoxicity of a test substance towards DNA repair-deficient CHO cell lines is compared with the corresponding cytotoxicity in the parental wild-type CHO cell line (AA8). A more pronounced toxicity toward a DNA repair-deficient cell line is interpreted as being the consequence of its inability to repair the DNA adduct induced by the compound. (+)-7beta,8alpha-Dihydroxy-9alpha,10alpha epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene, camptothecin, ethyl methanesulphonate and mitomycin C were used as reference substances, and the overall results indicate that the DRAG test could be useful in the screening of compounds for the production of repairable DNA adducts. The main advantages with the DRAG test are that it provides a relevant endpoint, it is rapid, it requires small amounts of the test item, and it permits a large number of compounds to be tested. PMID- 11387021 TI - Caspase-3 activity and carbonyl cyanide m-chlorophenylhydrazone-induced apoptosis in HL-60. AB - The role of caspase proteases in carbonyl cyanide m-chlorophenylhydrazone (CCCP) induced apoptosis of human promyelocytic HL-60 cells was examined. Treatment of HL-60 cells with micromolar concentrations of CCCP resulted in cell death, with typical apoptotic features such as chromatin condensation, formation of apoptotic bodies, nucleosomal fragmentation of DNA and a distinct increase in caspase-3 activity. The results, however, indicated that full caspase-3 inhibition by the selective inhibitor N-benzyloxycarbonyl-Asp-Glu-Val-Asp fluoromethyl ketone (Z DEVD-FMK) did not prevent cell death, nor did it affect the manifestation of apoptotic hallmarks, including apoptotic bodies formation and nucleosomal DNA fragmentation. The only distinct effect that Z-DEVD-FMK exhibited was to retard the disruption of the plasma membrane. We therefore assume that caspase-3 activity itself is not essential for the manifestation of apoptotic features mentioned above. Similarly, the pan-specific caspase inhibitor N benzyloxycarbonyl-Val-Ala-Asp fluoromethyl ketone (Z-VAD-FMK) did not prevent cell death. On the contrary, Z-VAD-FMK completely prevented DNA cleavage and apoptotic body formation, but it failed to completely counteract chromatin condensation. Thus, in the presence of Z-VAD-FMK, application of CCCP concentrations that otherwise induced apoptosis, resulted in the appearance of two morphologically different groups of dead cells with intact DNA. The first group included cells with necrotic-like nuclear morphology, and therefore could be taken as being "truly" necrotic in nature, because they had intact DNA. The cells of the second group formed small single-spherical nuclei with condensed chromatin. In spite of having intact DNA, they could not be taken as "truly" necrotic cells. It is evident that in the experimental system, caspase proteases play an essential role in the formation of apoptotic bodies and in the cleavage of nucleosomal DNA, but not in the condensation of chromatin. Therefore, it is likely that the choice between cell death modalities is not solely a matter of the caspase proteases present. PMID- 11387022 TI - Metabolic activity in primary cultures of fish hepatocytes. AB - In aquatic toxicology, isolated liver cells from fish can be used as a tool to generate initial information on the hepatic metabolism of xenobiotics, and on the mechanisms of xenobiotic activation or deactivation. This isolation of teleost liver cells is achieved by enzymic dissociation, and monolayer cultures of fish hepatocytes in serum-free medium maintain good viability for 3-8 days. During in vitro culture, fish liver cells express stable levels of phase I and phase II enzymes, such as cytochrome P4501A or glutathione S-transferase, and the cells show an induction of biotransformation enzymes after exposure to xenobiotics. The xenobiotic metabolite pattern produced by fish hepatocytes in vitro is generally similar to that observed in vivo. Limitations to more-intensive application of cultured fish hepatocytes as a screen in aquatic hazard assessment are partly due to the rather limited scope of existing studies, i.e. the focus on one particular species (rainbow trout), and on one particular biotransformation enzyme (cytochrome P4501A), as well as a lack of comparative in vitro/in vivo studies. PMID- 11387023 TI - Study of the environmental hazard caused by the oil shale industry solid waste. AB - The environmental hazard was studied of eight soil and solid waste samples originating from a region of Estonia heavily polluted by the oil shale industry. The samples were contaminated mainly with oil products (up to 7231mg/kg) and polycyclic aromatic hydrocarbons (PAHs; up to 434mg/kg). Concentrations of heavy metals and water-extractable phenols were low. The toxicities of the aqueous extracts of solid-phase samples were evaluated by using a battery of Toxkit tests (involving crustaceans, protozoa, rotifers and algae). Waste rock and fresh semi coke were classified as of "high acute toxic hazard", whereas aged semi-coke and most of the polluted soils were classified as of "acute toxic hazard". Analysis of the soil slurries by using the photobacterial solid-phase flash assay showed the presence of particle-bound toxicity in most samples. In the case of four samples out of the eight, chemical and toxicological evaluations both showed that the levels of PAHs, oil products or both exceeded their respective permitted limit values for the living zone (20mg PAHs/kg and 500mg oil products/kg); the toxicity tests showed a toxic hazard. However, in the case of three samples, the chemical and toxicological hazard predictions differed markedly: polluted soil from the Erra River bank contained 2334mg oil/kg, but did not show any water extractable toxicity. In contrast, spent rock and aged semi-coke that contained none of the pollutants in hazardous concentrations, showed adverse effects in toxicity tests. The environmental hazard of solid waste deposits from the oil shale industry needs further assessment. PMID- 11387024 TI - Cytotoxicity testing of wound-dressing materials. AB - A method was developed for testing the cytotoxicity of various bandage-like wound dressings and gel wound dressings. In this method, the ability of human polymorphonuclear neutrophils (PMNs) to initiate a respiratory burst after exposure to the various wound dressings is used as a marker of cytotoxicity. Luminol-amplified chemiluminescence stimulated with opsonised zymosan or phorbol 12-myristate 13-acetate (PMA) is used to measure the degree of activation of the respiratory burst, i.e. the NADPH oxidase activity, after exposure to wound dressings. Opsonised zymosan (material from yeast cell walls) is a phagocytic stimulus that activates the NADPH oxidase by binding to FC-receptors and complement receptors, and functions as an artificial bacterium, whereas PMA activates the NADPH oxidase by direct activation of protein kinase C. NADPH oxidase activity was inhibited by several wound dressings. The down-regulation of the respiratory burst is detrimental to the bactericial effect of PMNs, and can be used as a marker for the cytotoxicity of wound dressing materials. PMID- 11387025 TI - Biochemical responses in aquatic plants as markers of environmental contamination. AB - This short review gives several examples of the current status of xenobiotic biotransformation reactions and oxidative stress responses in plants as biomarkers of organic pollution in aquatic environments. Based on previous basic knowledge, several biomonitoring programmes have been applied successfully during the last decade. PMID- 11387026 TI - Protein precipitation In Vitro as a measure of chemical-induced cytotoxicity: an EDIT sub-programme. AB - As a priority area of the Evaluation-Guided Development of In Vitro Toxicity and Toxicokinetic Tests (EDIT) programme, an in vitro protein precipitation (PP) assay was used on the 50 reference chemicals of the Multicentre Evaluation of In Vitro Cytotoxicity (MEIC) project, to confirm and extend the MEIC results. Dose response curves were generated for only 30 of the chemicals, and the concentrations causing 10% (EC10) and 50% (EC50) protein precipitation versus the positive control were chosen as endpoints. The number of chemicals with a positive response increased to 46 when a new endpoint, the minimum effect concentration (MEC) that induces protein precipitation with respect to the negative control, was used. When the results were correlated with in vitro cytotoxicity in human cell lines, a similarly good correlation was found between the various endpoints of the PP assay at 5 hours and the 24-hour IC50 average cytotoxicity in human cell lines, even though the number of chemicals included in the correlation was larger for the MEC. Using the prediction error, the endpoint that gave the best correlation between the PP assay and human cell cytotoxicity was once more found to be the 5-hour MEC, and this was chosen for the PP assay. The sensitivity of the PP assay is lower than that of the in vitro cell-line cytotoxicity assay, possibly due to its shorter exposure period and because precipitation is the ultimate event in the sequence of a protein disturbance. It is expected that earlier denaturation steps would give better sensitivity. However, this simple, inexpensive and rapid assay could be useful in the early stages of testing chemicals. PMID- 11387027 TI - Uniform expression of alcohol dehydrogenase 3 in epithelia regenerated with cultured normal, immortalised and malignant human oral keratinocytes. AB - The human oral epithelium is a target for damage from the inhalation of formaldehyde. However, most experimental studies on this chemical have relied on laboratory animals that are obligatory nose breathers, including rats and mice. Therefore, in vitro model systems that mimic the structure of the human oral epithelium and which retain normal tissue-specific metabolic competence are desirable. Based on the established role of alcohol dehydrogenase 3 (ADH3), also known as glutathione-dependent formaldehyde dehydrogenase, as the primary enzyme catalysing the detoxification of formaldehyde, the aim of this study was to investigate the expression of ADH3 in organotypic epithelia regenerated with normal (NOK), immortalised (SVpgC2a) and malignant (SqCC/Y1) human oral keratinocytes. Organotypic epithelia, usually consisting of 5-10 cell layers, were produced at the air-liquid interface of collagen gels containing human oral fibroblasts, after culture for 10 days in a standardised serum-free medium. Immunochemical staining demonstrated uniform expression of ADH3 in these organotypic epithelia, as well as in the epithelial cells of oral tissue. The specificity of the ADH3 antiserum was ascertained from the complete neutralisation of the immunochemical reaction with purified ADH3 protein. Assessment of the staining intensities indicated that the expression levels were similar among the regenerated epithelia. Furthermore, the regenerated epithelia showed similar ADH3 expression to the epithelium in oral tissue. Therefore, a tissue-like expression pattern for ADH3 can be generated from the culture of various oral keratinocyte lines in an organotypic state. Similar expression levels among the various cell lines indicate the preservation of ADH3 during malignant transformation, and therefore that NOK, SVpgC2a and SqCC/Y1 represent functional models for in vitro studies of formaldehyde metabolism in human oral mucosa. PMID- 11387028 TI - Pyruvate-induced long-term maintenance of glutathione s-transferase in rat hepatocyte cultures. AB - The addition of pyruvate to the culture medium has been reported to improve the maintenance of P450-dependent enzyme expression in primary rat hepatocyte cultures. In this study, the effects of 30mM pyruvate on cell morphology, albumin secretion and glutathione S-transferase (GST) expression were investigated as a function of the time in culture. The effect of triiodothyronine (T3) exposure on GST expression was also measured in pyruvate-treated cultures. Transmission electron microscopy showed that untreated hepatocytes deteriorated after culture for 7 days, whereas the morphology of the pyruvate-treated cells was similar to that observed in intact liver tissue. The albumin secretion rate was significantly higher in rat hepatocytes exposed to pyruvate than in control cells. In the presence of pyruvate, mu and alpha class GST activities were well maintained, whereas GST pi activity was increased over the entire culture period. HPLC analysis revealed that the complement of GST subunits present in hepatocytes is altered during culture with pyruvate: mu,class proteins remained relatively constant, whereas a decrease in the alpha class content was accompanied by a strong increase in GST subunit P1 (GSTP1). The induction of GSTP1 was confirmed at the mRNA level. In control cultures, pi class GST activity was increased, but total, mu, and alpha class GST activities continuously declined as a function of culture time and became undetectable beyond 7 days in culture. At the protein and mRNA levels, a much smaller increase in GSTP1 was observed than in the pyruvate cultures. When the pyruvate-treated cell cultures were exposed to T3, an inhibitory effect on GST activities and proteins was found. These results indicate that this simple culture model could be useful for studying the expression and regulation of GST. PMID- 11387029 TI - A compartment model to calculate time-dependent concentration profiles of topically applied chemical compounds in the anterior compartments of the rabbit eye. AB - Hitherto, none of the existing in vitro methods has been convincingly demonstrated to be suitable as a replacement for the Draize rabbit eye irritation test. We examine the hypothesis that one reason for this is that insufficient consideration has been given to the differences in the effective concentrations at which chemicals operate in vitro and in vivo. When a chemical is applied topically to the eye, the strength of the observed irritation that it elicits depends both on its toxic potential toward cells or tissues, and its effective concentration in the tissues of the eye. Most of the existing in vitro methods are based on isolated cells or tissues, and thus may be useful in assessing the cytotoxic potentials of chemicals. However, a reliable approach to assessing the effective concentrations of chemicals within the various tissues of the eye is lacking. A simplified compartment model is presented for calculating the time dependent, intra-ocular concentration profiles of topically applied chemicals. The model encompasses the outer surface of the eye, three distinct segments of the cornea (subdivided into the epithelium, stroma and endothelium) and the conjunctiva. Transport through the membranes of these compartments is described as passive diffusion. For the transport coefficients, rate equations are established that contain, as free parameters, the molecular size and the partition coefficient of the chemical, as well as some intrinsic membrane parameters, such as thickness, viscosity and pore density. Numerical values for the unknown membrane parameters were estimated by fitting the theoretical rate equations to measured permeability coefficients. The compartment model was applied to an independent set of 52 test chemicals compiled from the European Commission/UK Home Office validation study. The calculated passage times (required to let 95% of the chemical reach the posterior eye tissues) varied between 0.33 minutes and 50.6 minutes, and are generally much shorter than the typical duration of observed impairments in the cornea or conjunctiva. This finding suggests that short-term contacts of the eye tissues with a chemical are sufficient to elicit long-term eye irritation. An example is given, showing how the conventional approach of using in vitro endpoints as predictors of eye irritation can be improved significantly by incorporating into the prediction the calculated intra-ocular concentration of a chemical. PMID- 11387031 TI - Effects of Cu2+, Ni2+, Pb2+, Zn2+ and pentachlorophenol on photosynthesis and motility in Chlamydomonas reinhardtii in short-term exposure experiments. AB - BACKGROUND: Heavy metals, especially copper, nickel, lead and zinc, have adverse effects on terrestrial and in aquatic environments. However, their impact can vary depending on the nature of organisms. Taking into account the ability of heavy metals to accumulate in sediments, extended knowledge of their effects on aquatic biota is needed. In this context the use of model organisms (often unicellular), which allows for rapid assessment of pollutants in freshwater, can be of advantage. Pentachlorophenol has been extensively used for decades as a bleaching agent by pulp- and paper industry. Pentachlorophenol tends to accumulate in the nature. We aim to determine if photosynthesis and motility can be used as sensitive physiological parameters in toxicological studies of Chlamydomonas reinhardtii, a motile green unicellular alga. It is discussed if photosynthesis and motility can be used as sensitive physiological parameters in toxicological studies. RESULTS: The concentrations studied ranged from 0.1 to 2.0 mg l-1 for copper, nickel, lead and zinc, and from 0.1 to 10.0 mg l-1 for pentachlorophenol. Exposure time was set to 24 h. Copper and pentachlorophenol turned out to be especially toxic for photosynthetic efficiency (PE) in C. reinhardtii. CONCLUSION: Copper and pentachlorophenol turned out to be especially toxic for PE in C. reinhardtii. Zinc has been concluded to be moderately toxic while nickel and lead had stimulatory effects on the PE. Because of high variance, motility was not considered a reliable physiological parameter when assessing toxicity of the substances using C. reinhardtii. PMID- 11387032 TI - After the (draft) sequence. PMID- 11387033 TI - The Rosetta stone. PMID- 11387034 TI - Microarray gene expression database: progress towards an international repository of gene expression data. PMID- 11387035 TI - Macromolecular technologies: applications and improvements. PMID- 11387037 TI - Identification of conserved C2H2 zinc-finger gene families in the Bilateria. AB - BACKGROUND: Identification of orthologous relationships between genes from widely divergent taxa allows partial reconstruction of the gene complement of ancestral genomes. C2H2 zinc-finger genes are one of the largest and most complex gene superfamilies in metazoan genomes, with hundreds of members in the human genome. Here we analyze C2H2 zinc-finger genes from three taxa - Drosophila, Caenorhabditis elegans and human - from which near-complete genome sequence data are available. RESULTS: Our analyses conclusively identify 39 families of genes, of which 38 can be defined as orthology groups in that they are descended from single ancestral genes in the common ancestor of Drosophila, C. elegans and humans. CONCLUSIONS: On the basis of current metazoan phylogeny, these 39 groups represent the minimum complement of C2H2 zinc-finger genes present in the genome of the bilaterian common ancestor. PMID- 11387036 TI - Identification of candidate downstream genes for the homeodomain transcription factor Labial in Drosophila through oligonucleotide-array transcript imaging. AB - BACKGROUND: Homeotic genes are key developmental regulators that are highly conserved throughout evolution. Their encoded homeoproteins function as transcription factors to control a wide range of developmental processes. Although much is known about homeodomain-DNA interactions, only a small number of genes acting downstream of homeoproteins have been identified. Here we use a functional genomic approach to identify candidate target genes of the Drosophila homeodomain transcription factor Labial. RESULTS: High-density oligonucleotide arrays with probe sets representing 1,513 identified and sequenced genes were used to analyze differential gene expression following labial overexpression in Drosophila embryos. We find significant expression level changes for 96 genes belonging to all functional classes represented on the array. In accordance with our experimental procedure, we expect that these genes are either direct or indirect targets of labial gene action. Among these genes, 48 were upregulated and 48 were downregulated following labial overexpression. This corresponds to 6.3% of the genes represented on the array. For a selection of these genes, we show that the data obtained with the oligonucleotide arrays are consistent with data obtained using quantitative RT-PCR. CONCLUSIONS: Our results identify a number of novel candidate downstream target genes for Labial, suggesting that this homeoprotein differentially regulates a limited and distinct set of embryonically expressed Drosophila genes. PMID- 11387038 TI - Genomic structure of the gene for mouse germ-cell nuclear factor (GCNF). II. Comparison with the genomic structure of the human GCNF gene. AB - BACKGROUND: Germ-cell nuclear factor (GCNF, NR6AI) is an orphan nuclear receptor. Its expression pattern suggests it functions during embryogenesis, in the placenta and in germ-cell development. Mouse GCNF cDNA codes for a protein of 495 amino acids, whereas the four reported human cDNA variants code for proteins of 454 to 480 amino acids. Apart from this size difference, there is sequence conservation of up to 98.7%. To elucidate the genomic structure that gives rise to the different human GCNF mRNAs, the sequence information of the human GCNF locus is compared to the previously reported structure of the mouse locus. RESULTS: The genomic structures of the mouse and human GCNF genes are highly conserved. The comparison reveals that the shorter human protein results from skipping the 45 base-pair third exon. Three different human isoforms - GCNF-1, GCNF-2a and GCNF-2b - are generated by differential usage of alternative splice acceptor sites of the fourth and the seventh exon. CONCLUSION: By homology with the mouse gene, 11 GCNF coding exons can be defined on human chromosome 9. All human GCNF cDNAs identified so far are, however, derived from mRNAs generated by splicing the fourth to the second exon. Although the genomic sequence is highly conserved, the analysis suggests that alternative splicing generates a higher complexity of human GCNF isoforms compared with the situation in the mouse. PMID- 11387039 TI - Characterizing glycosylation pathways. AB - Numerous factors that influence cell-surface carbohydrate composition remain to be elucidated. The combination of novel biochemical and metabolism-based approaches with emerging genomic methods promises to accelerate efforts to understand glycosylation. PMID- 11387040 TI - Homologous recombination: from model organisms to human disease. AB - Recent experiments show that properly controlled recombination between homologous DNA molecules is essential for the maintenance of genome stability and for the prevention of tumorigenesis. PMID- 11387041 TI - Morpholino antisense oligonucleotides: tools for investigating vertebrate development. AB - Antisense oligonucleotides provide a promising approach to investigating gene function in vivo, but their ability to offer unambiguous insights into phenotypes has been debated. The recent use of morpholino antisense oligonucleotides in zebrafish embryos may prove a major advance, but rigorous controls are essential. PMID- 11387042 TI - Towards a complete sequence of the human Y chromosome. AB - A few dozen genes are known on the human Y chromosome. The completion of the human genome sequence will allow identification of the remaining loci, which should shed further light on the function and evolution of this peculiar chromosome. PMID- 11387044 TI - Generalization in interactive networks: the benefits of inhibitory competition and Hebbian learning. AB - Computational models in cognitive neuroscience should ideally use biological properties and powerful computational principles to produce behavior consistent with psychological findings. Error-driven backpropagation is computationally powerful and has proven useful for modeling a range of psychological data but is not biologically plausible. Several approaches to implementing backpropagation in a biologically plausible fashion converge on the idea of using bidirectional activation propagation in interactive networks to convey error signals. This article demonstrates two main points about these error-driven interactive networks: (1) they generalize poorly due to attractor dynamics that interfere with the network's ability to produce novel combinatorial representations systematically in response to novel inputs, and (2) this generalization problem can be remedied by adding two widely used mechanistic principles, inhibitory competition and Hebbian learning, that can be independently motivated for a variety of biological, psychological, and computational reasons. Simulations using the Leabra algorithm, which combines the generalized recirculation (GeneRec), biologically plausible, error-driven learning algorithm with inhibitory competition and Hebbian learning, show that these mechanisms can result in good generalization in interactive networks. These results support the general conclusion that cognitive neuroscience models that incorporate the core mechanistic principles of interactivity, inhibitory competition, and error-driven and Hebbian learning satisfy a wider range of biological, psychological, and computational constraints than models employing a subset of these principles. PMID- 11387043 TI - The Rab GTPase family. AB - SUMMARY: The Rab family is part of the Ras superfamily of small GTPases. There are at least 60 Rab genes in the human genome, and a number of Rab GTPases are conserved from yeast to humans. The different Rab GTPases are localized to the cytosolic face of specific intracellular membranes, where they function as regulators of distinct steps in membrane traffic pathways. In the GTP-bound form, the Rab GTPases recruit specific sets of effector proteins onto membranes. Through their effectors, Rab GTPases regulate vesicle formation, actin- and tubulin-dependent vesicle movement, and membrane fusion. PMID- 11387045 TI - Optimal smoothing in visual motion perception. AB - When a flash is aligned with a moving object, subjects perceive the flash to lag behind the moving object. Two different models have been proposed to explain this "flash-lag" effect. In the motion extrapolation model, the visual system extrapolates the location of the moving object to counteract neural propagation delays, whereas in the latency difference model, it is hypothesized that moving objects are processed and perceived more quickly than flashed objects. However, recent psychophysical experiments suggest that neither of these interpretations is feasible (Eagleman & Sejnowski, 2000a, 2000b, 2000c), hypothesizing instead that the visual system uses data from the future of an event before committing to an interpretation. We formalize this idea in terms of the statistical framework of optimal smoothing and show that a model based on smoothing accounts for the shape of psychometric curves from a flash-lag experiment involving random reversals of motion direction. The smoothing model demonstrates how the visual system may enhance perceptual accuracy by relying not only on data from the past but also on data collected from the immediate future of an event. PMID- 11387046 TI - Rate coding versus temporal order coding: what the retinal ganglion cells tell the visual cortex. AB - It is often supposed that the messages sent to the visual cortex by the retinal ganglion cells are encoded by the mean firing rates observed on spike trains generated with a Poisson process. Using an information transmission approach, we evaluate the performances of two such codes, one based on the spike count and the other on the mean interspike interval, and compare the results with a rank order code, where the first ganglion cells to emit a spike are given a maximal weight. Our results show that the rate codes are far from optimal for fast information transmission and that the temporal structure of the spike train can be efficiently used to maximize the information transfer rate under conditions where each cell needs to fire only one spike. PMID- 11387047 TI - The effects of spike frequency adaptation and negative feedback on the synchronization of neural oscillators. AB - There are several different biophysical mechanisms for spike frequency adaptation observed in recordings from cortical neurons. The two most commonly used in modeling studies are a calcium-dependent potassium current I(ahp) and a slow voltage-dependent potassium current, I(m). We show that both of these have strong effects on the synchronization properties of excitatorily coupled neurons. Furthermore, we show that the reasons for these effects are different. We show through an analysis of some standard models, that the M-current adaptation alters the mechanism for repetitive firing, while the afterhyperpolarization adaptation works via shunting the incoming synapses. This latter mechanism applies with a network that has recurrent inhibition. The shunting behavior is captured in a simple two-variable reduced model that arises near certain types of bifurcations. A one-dimensional map is derived from the simplified model. PMID- 11387048 TI - A unified approach to the study of temporal, correlational, and rate coding. AB - We demonstrate that the information contained in the spike occurrence times of a population of neurons can be broken up into a series of terms, each reflecting something about potential coding mechanisms. This is possible in the coding regime in which few spikes are emitted in the relevant time window. This approach allows us to study the additional information contributed by spike timing beyond that present in the spike counts and to examine the contributions to the whole information of different statistical properties of spike trains, such as firing rates and correlation functions. It thus forms the basis for a new quantitative procedure for analyzing simultaneous multiple neuron recordings and provides theoretical constraints on neural coding strategies. We find a transition between two coding regimes, depending on the size of the relevant observation timescale. For time windows shorter than the timescale of the stimulus-induced response fluctuations, there exists a spike count coding phase, in which the purely temporal information is of third order in time. For time windows much longer than the characteristic timescale, there can be additional timing information of first order, leading to a temporal coding phase in which timing information may affect the instantaneous information rate. In this new framework, we study the relative contributions of the dynamic firing rate and correlation variables to the full temporal information, the interaction of signal and noise correlations in temporal coding, synergy between spikes and between cells, and the effect of refractoriness. We illustrate the utility of the technique by analyzing a few cells from the rat barrel cortex. PMID- 11387050 TI - Attractive periodic sets in discrete-time recurrent networks (with emphasis on fixed-point stability and bifurcations in two-neuron networks). AB - We perform a detailed fixed-point analysis of two-unit recurrent neural networks with sigmoid-shaped transfer functions. Using geometrical arguments in the space of transfer function derivatives, we partition the network state-space into distinct regions corresponding to stability types of the fixed points. Unlike in the previous studies, we do not assume any special form of connectivity pattern between the neurons, and all free parameters are allowed to vary. We also prove that when both neurons have excitatory self-connections and the mutual interaction pattern is the same (i.e., the neurons mutually inhibit or excite themselves), new attractive fixed points are created through the saddle-node bifurcation. Finally, for an N-neuron recurrent network, we give lower bounds on the rate of convergence of attractive periodic points toward the saturation values of neuron activations, as the absolute values of connection weights grow. PMID- 11387049 TI - Determination of response latency and its application to normalization of cross correlation measures. AB - It is often of interest experimentally to assess how synchronization between two neurons changes following a stimulus or other behaviorally relevant marker. The joint peristimulus time histogram (JPSTH) achieves this, but assumes that changes in the cells' firing rate following the stimulus are stereotyped from one sweep to the next. Erroneous results can be generated if this is not the case. We here present a method to assess whether there are variations in response latency or amplitude from sweep to sweep. We then describe how the effects of response latency variation can be mitigated by realigning sweeps to their individual latencies. Three methods of detecting response latency are presented and their performance compared on simulated data. Finally, the effect on the JPSTH of sweep realignment using detected latencies is illustrated. PMID- 11387051 TI - Attractor networks for shape recognition. AB - We describe a system of thousands of binary perceptrons with coarse-oriented edges as input that is able to recognize shapes, even in a context with hundreds of classes. The perceptrons have randomized feedforward connections from the input layer and form a recurrent network among themselves. Each class is represented by a prelearned attractor (serving as an associative hook) in the recurrent net corresponding to a randomly selected subpopulation of the perceptrons. In training, first the attractor of the correct class is activated among the perceptrons; then the visual stimulus is presented at the input layer. The feedforward connections are modified using field-dependent Hebbian learning with positive synapses, which we show to be stable with respect to large variations in feature statistics and coding levels and allows the use of the same threshold on all perceptrons. Recognition is based on only the visual stimuli. These activate the recurrent network, which is then driven by the dynamics to a sustained attractor state, concentrated in the correct class subset and providing a form of working memory. We believe this architecture is more transparent than standard feedforward two-layer networks and has stronger biological analogies. PMID- 11387052 TI - Nonviral vectors in the new millennium: delivery barriers in gene transfer. AB - Development of an efficient method for introducing a therapeutic gene into target cells in vivo is the key issue in treating genetic and acquired diseases by gene therapy. To this end, various nonviral vectors have been designed and developed, and some of them are in clinical trials. The simplest approach is naked DNA injection into local tissues or systemic circulation. Physical (gene gun, electroporation) and chemical (cationic lipid or polymer) approaches have also been utilized to improve the efficiency and target cell specificity of gene transfer by plasmid DNA. After administration, however, nonviral vectors encounter many hurdles that result in diminished gene transfer in target cells. Cationic vectors sometimes attract serum proteins and blood cells when entering into blood circulation, which results in dynamic changes in their physicochemical properties. To reach target cells, nonviral vectors should pass through the capillaries, avoid recognition by mononuclear phagocytes, emerge from the blood vessels to the interstitium, and bind to the surface of the target cells. They then need to be internalized, escape from endosomes, and then find a way to the nucleus, avoiding cytoplasmic degradation. Successful clinical applications of nonviral vectors will rely on a better understanding of barriers in gene transfer and development of vectors that can overcome these barriers. PMID- 11387053 TI - In vivo gene transfer in mouse skeletal muscle mediated by baculovirus vectors. AB - Baculovirus vectors are efficient tools for gene transfer into mammalian cells in vitro. However, in vivo gene delivery by systemic administration is hindered by the vector inactivation mediated by the complement system. To characterize further the gene transfer efficacy of baculovirus we examined the vector transduction efficiency in skeletal muscle. Vectors expressing vesicular stomatitis virus glycoprotein (VSV-G) in the viral envelope were generated by inserting the VSV-G coding sequence downstream of the polyhedrin promoter. Two viruses were constructed to carry either the Escherichia coli beta-galactosidase (beta-Gal) gene or the mouse erythropoietin (EPO) cDNA cloned downstream of the cytomegalovirus immediate-early promoter and enhancer. The greater gene transduction efficiency of the Bac-G-betaGal vector was confirmed by comparing the beta-Gal expression level in a variety of human and mouse cell lines with that obtained on infection with Bac-betaGal, a vector that lacks VSV-G. Similarly, a 5- to 10-fold increase in beta-Gal expression between Bac-G-betaGal and Bac-betaGal was observed when mouse myoblasts and myotubes were infected. The same increase in beta-Gal expression was detected on injection of the Bac-G betaGal vector in the quadriceps of BALB/c and C57BL/6 mice. In contrast, a 2 fold difference in transduction was observed between these two vectors in DBA/2J mouse strain. Last, expression of EPO cDNA was detected for at least 178 days in DBA/2J mice on Bac-G-EPO injection into the quadriceps whereas EPO expression declined to normal values by 35 days postinfection in BALB/c and C57BL/6 mice. Thus, these results indicate that baculovirus may be considered a useful vector for gene transfer in mouse skeletal muscle and that persistence of expression may depend on the mouse strain used. PMID- 11387054 TI - Interleukin 2 gene therapy for prostate cancer: phase I clinical trial and basic biology. AB - Twenty-four patients with locally advanced prostate cancer (CaP) were enrolled in a phase I clinical trial using gene-based immunotherapy. A functional DNA-lipid complex encoding the interleukin 2 (IL-2) gene (Leuvectin; Vical, San Diego, CA) was administered intraprostatically into the hypoecogenic tumor lesion, using transrectal ultrasound guidance. Two groups of patients having locally advanced tumors were enrolled to receive a treatment regimen composed of two serial intraprostatic injections of the IL-2 gene agent administered 1 week apart. The first groups of patients included radical prostatectomy candidates who subsequently underwent surgery after the completion of the treatment regimen. The second group consisted of patients who had failed a prior therapy. Prostate specimens of the treated areas were attained after treatment and compared with the transrectal biopsies performed at baseline to assess for any responses. IL-2 gene therapy was well tolerated, with no grade 3 or 4 toxic reactions occurring. The most commonly reported symptoms were mild hematuria, transient rectal bleeding, and perineal discomfort that are likely attributable to the injection itself. During the entire course of treatment, there were no significant changes in American Urologic Association (AUA) symptom scores, in hematologic disturbances, electrolyte imbalances, or hepatic functions. Evidence of systemic immune activation was observed after IL-2 gene therapy, based on an increase in the intensity of T cell infiltration seen on immunohistochemical analysis of tissue samples from the injected tumor sites, and based on increased proliferation rates of peripheral blood lymphocytes that were cocultured with patient serum collected after treatment. Furthermore, transient decreases in serum prostate-specific antigen (PSA) (responders) were seen in 16 of 24 patients (67%) on day 1. Fourteen of the patients persisted in this decrease to day 8 (58%). In eight patients the PSA level rose (nonresponders). More patients (9 to 10) in the group that failed prior therapy responded to the IL-2 gene injections (chi-square test, p = 0.04), and 6 of the 9 also had lower than baseline PSA levels at week 10 after treatment. To the best of our knowledge, this is the first clinical study of its kind aimed at exploring the role of IL-2-based gene therapy in CaP patients. This phase I trial demonstrated the safety of intraprostatic Leuvectin injection, with transient PSA-based responses seen after therapy. PMID- 11387055 TI - Intracardiac echocardiographic guidance and monitoring during percutaneous endomyocardial gene injection in porcine heart. AB - In an effort to develop a guiding and monitoring tool for transmyocardial gene transfer, we have evaluated the feasibility of intracardiac echocardiography (ICE) to guide percutaneous endomyocardial gene transfer (PEGT), and monitor complications, in a pig model. ICE (5.5-10 MHz), complemented by fluoroscopy, was utilized to guide a needle injection into the heart in 19 normal pigs. Using this system, we injected Evans blue dye into eight pigs (group I), a mixture of pCK CAT plasmid and India ink into seven pigs (group II), and pCK-LacZ plasmid into four pigs (group III). In all pigs, ICE contributed to the injection procedure by guiding the catheter to anatomically distinct sites, and by assisting stabilization of the catheter-endocardial contact. ICE predicted the injection sites correctly in 56 of 64 sites (87.5%) in group I, and in 42 of 42 sites (100%) in group II. Leakage of injectate into the left ventricular cavity could be detected by the microbubbles generated. The sites of injections appeared as foci of bright myocardial echodensity, which persisted until the end of the procedure. The procedures were not associated with significant morbidity or mortality. The expression of the chloramphenicol acetyltransferase (CAT) gene was identified in 40 sites from 42 injections (95.2%) in group II. In group III, histology showed positive beta-galactosidase staining of myocytes limited around the needle track with low transfection efficiency (<1%). These results suggest that real-time ICE monitoring proves safe and useful during PEGT for guiding needle injection, monitoring leakage, ensuring delivery of injectate into the myocardium, and instantly diagnosing cardiac complications, resulting in successful gene transfer. PMID- 11387056 TI - Delivery to the central nervous system of a nonreplicative herpes simplex type 1 vector engineered with the interleukin 4 gene protects rhesus monkeys from hyperacute autoimmune encephalomyelitis. AB - Systemic administration of antiinflammatory molecules to patients affected by immune-mediated inflammatory demyelinating diseases of the central nervous system (CNS) has limited therapeutic efficacy due to the presence of the blood-brain barrier (BBB). We found that three of five rhesus monkeys injected intrathecally with a replication-defective herpes simplex virus (HSV) type 1-derived vector engineered with the human interleukin 4 (IL-4) gene were protected from an hyperacute and lethal form of experimental autoimmune encephalomyelitis induced by whole myelin. The intrathecally injected vector consistently diffused within the CNS via the cerebrospinal fluid and infected ependymal cells, which in turn sustained in situ production of IL-4 without overt immunological or toxic side effects. In EAE-protected monkeys, IL-4-gene therapy significantly decreased the number of brain as well as spinal cord inflammatory perivenular infiltrates and the extent of demyelination, necrosis, and axonal loss. The protective effect was associated with in situ downregulation of inflammatory mediators such as tumor necrosis factor alpha (TNF-alpha) and monocyte chemoattractant protein 1 (MCP-1), upregulation of transforming growth factor beta (TGF-beta), and preservation of BBB integrity. Our results indicate that intrathecal delivery of HSV-1-derived vectors containing antiinflammatory cytokine genes may play a major role in the future therapeutic armamentarium of inflammatory CNS-confined demyelinating diseases and, in particular, in the most fulminant forms where conventional therapeutic approaches have, so far, failed to achieve a satisfactory control of the disease evolution. PMID- 11387057 TI - A uniquely stable replication-competent retrovirus vector achieves efficient gene delivery in vitro and in solid tumors. AB - A major obstacle in cancer gene therapy is the limited efficiency of in vivo gene transfer by replication-defective retrovirus vectors in current use. One strategy for circumventing this difficulty would be to use vectors capable of replication within tumor tissues. We have developed a replication-competent retrovirus (RCR) vector derived from murine leukemia virus (MuLV). This vector utilizes a unique design strategy in which an internal ribosome entry site-transgene cassette is positioned between the env gene and the 3' long terminal repeat (LTR). The ability of this vector to replicate and transmit a transgene was examined in culture and in a solid tumor model in vivo. The RCR vector exhibited replication kinetics similar to those of wildtype MuLV and mediated efficient delivery of the transgene throughout an entire population of cells in culture after an initial inoculation with 1 plaque-forming unit (PFU) of vector per 2000 cells. After injection of 6 x 10(3) PFU of vector into established subcutaneous tumors, highly efficient spread of the transgene was observed over a period of 7 weeks, in some cases resulting in spread of the transgene throughout the entire tumor. MuLV based RCR vectors show significant advantages over standard replication-defective vectors in efficiency of gene delivery both in culture and in vivo. This represents the first example of the use of an RCR vector in an adult mammalian host, and their first application to transduction of solid tumors. PMID- 11387058 TI - Evaluation of retroviral vector design in defined chromosomal loci by Flp mediated cassette replacement. AB - Successful retroviral vector construction is still empirical. Test systems for vector efficiency are based on statistical comparison of numerous infectants with single proviral integrates, since their expression depends on the chromosomal surroundings. More reliable data would be obtained if different vector constructs were studied in an identical chromosomal context. Here, we demonstrate the use of a new method, in which chromosomal sites are provirally tagged in such a way that they can be targeted with other expression cassettes. The original tagging integrate is replaced in one step by the targeting element. This permits a reliable comparison of different retroviral vector configurations, eliminating the influence of neighboring chromosomal elements. We compared different retroviral vector types for coexpression of two genes: a vector containing an internal promoter and a vector with an internal ribosome entry site (IRES) element. In contrast to bicistronic retroviral vectors, dual-promoter proviruses exhibited rapid inactivation of the long terminal repeat (LTR)-driven gene expression. Targeted exchange of the dual-promoter provirus with a bicistronic retroviral cassette resulted in gain of expression stability. The reverse experiment confirmed this promoter interaction phenomenon since initial expression stability from a single-promoter bicistronic provirus was lost by targeted exchange with a dual-promoter cassette. In addition, targeting exchange of the dual-promoter provirus, replacing the LTR with an artificial (Tet) promoter restored expression stability. These observations, valid for various integration sites, prove the strong interaction between the LTR and the internal promoter. Our results have implications for retroviral vector design and suggest that retroviral coexpression of two genes is more predictable in the bicistronic configuration. PMID- 11387059 TI - An episomally maintained MDR1 gene for gene therapy. AB - Potential applications of the MDR1 multidrug transporter in gene therapy include protecting sensitive bone marrow cells against cytotoxic drugs during cancer chemotherapy and serving as a dominant selectable marker when coexpressed with a corrective passenger gene. To address safety concerns associated with integrating viral systems, such as retroviruses, we tested the feasibility of maintaining a nonvirally delivered MDR1 gene (pEpiHaMA) episomally. An MDR1 vector containing the Epstein-Barr virus (EBV) origin of replication (OriP) and its nuclear retention protein (EBNA-1) was transfected into human (KB-3-1) cells. MDR1 was expressed at a higher level in cells carrying the episomal vector, pEpiHaMA, compared with the vector lacking sequences needed for episomal maintenance (pHaMA). Furthermore, more drug-resistant KB-3-1 colonies were obtained on selection after transfection with pEpiHaMA. These observations correlated with longer maintenance of episomes in cells transfected with pEpiHaMA. In addition, episomes could still be recovered for more than 1 month from tumor explants in nude mice that were injected with pEpiHaMA-liposome complexes after drug selection, suggesting that these constructs can be maintained extrachromosomally in vivo. PMID- 11387060 TI - Long-term efficacy after [E1-, polymerase-] adenovirus-mediated transfer of human acid-alpha-glucosidase gene into glycogen storage disease type II knockout mice. AB - Glycogen storage disease type II (GSD-II) is a lethal, autosomal recessive metabolic myopathy caused by a lack of acid-alpha-glucosidase (GAA) activity in the cardiac and skeletal muscles. Absence of adequate intralysosomal GAA activity results in massive amounts of glycogen accumulation in multiple muscle groups, resulting in morbidity and mortality secondary to respiratory embarrassment and/or cardiomyopathy. In a mouse model of GSD-II, we demonstrate that infection of the murine liver with a modified adenovirus (Ad) vector encoding human GAA (hGAA) resulted in long-term persistence of the vector in liver tissues for at least 6 months. Despite both a rapid shutdown of hGAA mRNA expression from the vector, as well as the elicitation of anti-hGAA antibody responses (hGAA is a foreign antigen in this model), the hGAA secreted by the liver was taken up by all muscle groups analyzed and, remarkably, persisted in them for at least 6 months. The persistence of the protein also correlated with long-term correction of pathologic intramuscular glycogen accumulations in all muscle groups tested, but most notably the cardiac tissues, which demonstrated a significantly decreased glycogen content for at least 190 days after a single vector injection. The results suggest that gene therapy strategies may have the potential to significantly improve the clinical course for GSD-II patients. PMID- 11387061 TI - Transfer of heme oxygenase 1 cDNA by a replication-deficient adenovirus enhances interleukin 10 production from alveolar macrophages that attenuates lipopolysaccharide-induced acute lung injury in mice. AB - By using a direct, intratracheal inoculation of an adenovirus encoding heme oxygenase 1 (Ad.HO-1), model gene therapy for acute lung injury induced by inhaled pathogen was performed. Data demonstrated that Ad.HO-1 administration is as effective as the pharmacologic upregulation of the endogenous HO-1 gene expression by hemin to attenuate neutrophilic inflammations of the lung after aerosolized lipopolysaccharide (LPS) exposure. Interestingly, immunohistochemical analysis revealed that the HO-1 gene was transferred not only to the airway epithelium, but to the alveolar macrophages (AMs). Moreover, overexpression of exogenous HO-1 in the macrophages provided a high level of endogenous interleukin 10 (IL-10) production from the macrophages, and additional experiments using IL 10 knockout mice demonstrated that the increase in IL-10 in the macrophages was critical for the resolution of neutrophilic migration in the lung after LPS exposure. These results suggest that AMs not only are barriers for efficient gene transfer to the respiratory epithelium, but also represent logical targets for Ad mediated, direct, in vivo gene therapy strategies for inflammatory disorders in humans. PMID- 11387063 TI - Preclinical safety evaluation of G207, a replication-competent herpes simplex virus type 1, inoculated intraprostatically in mice and nonhuman primates. AB - G207, a replication-competent herpes simplex virus type 1 (HSV-1) virus, has been previously shown to be effective against human prostate cancer xenografts in mice. This study assesses its safety in the prostate of two animal models known for their sensitivity to HSV-1. BALB/c mice were injected intraprostatically with either HSV-1 G207 or strain F and observed for 5 months. None of the G207 injected animals exhibited any clinical signs of disease or died. However, 50% of strain F-injected mice displayed sluggish, hunched behavior and died by day 13. Histopathologically, the G207-injected prostates were normal whereas strain F injected prostates showed epithelial flattening, sloughing, and stromal edema. Four Aotus nancymae monkeys were also injected with G207 intraprostatically and observed short term (up to 21 days) and long term (56 days). Safety was assessed on the basis of clinical observations, viral biodistribution, virus shedding, and histopathology. None of the injected monkeys displayed evidence of clinical disease, shedding of infectious virus, or spread of the virus into other organs. Except for minor histological changes unrelated to the study, no significant abnormalities were observed. These results demonstrate that G207 can be safely inoculated into the prostate and should be considered for human trials for the treatment of prostate cancer. PMID- 11387062 TI - A new-generation stable inducible packaging cell line for lentiviral vectors. AB - We have successfully generated and characterized a stable packaging cell line for HIV-1-based vectors. To allow safe production of vector, a minimal packaging construct carrying only the coding sequences of the HIV-1 gag-pol, tat, and rev genes was stably introduced into 293G cells under the control of a Tet(o) minimal promoter. 293G cells express the chimeric Tet(R)/VP16 trans-activator and contain a tetracycline-regulated vesicular stomatitis virus protein G (VSV-G) envelope gene. When the cells were grown in the presence of tetracycline the expression of both HIV-1-derived and VSV-derived packaging functions was suppressed. On induction, approximately 50 ng/ml/24 hr of Gag p24 equivalent of vector was obtained. After introduction of the transfer vector by serial infection, vector could be collected for several days with a transduction efficiency similar or superior to that of vector produced by transient transfection both for dividing and growth-arrested cells. The vector could be effectively concentrated to titers reaching 10(9) transducing units/ml and allowed for efficient delivery and stable expression of a GFP transgene in the mouse brain. The packaging cell line and all vector producer clones described here were shown to be free from replication competent recombinants, and from recombinants between packaging and vector constructs that transfer the viral gag-pol genes. The packaging cell line and the assays developed will advance lentiviral vectors toward the stringent requirements of clinical applications. PMID- 11387065 TI - Percutaneous Biopsy of Musculoskeletal Lesions. AB - Percutaneous biopsy is a minimally invasive technique that yields a high diagnostic accuracy at a relatively low cost in musculoskeletal lesions, especially tumors and infection. In this article, technical refinements improving the results of percutaneous biopsy of musculoskeletal lesions are reviewed, especially the type of radiological guidance, the different biopsy needles available, and the technique of approach depending on the lesion site. Reported complications, results, and indications of musculoskeletal percutaneous biopsy are also discussed. PMID- 11387066 TI - Percutaneous Biopsy of the Synovial Membrane. AB - Techniques of percutaneous synovial biopsy under fluoroscopic control of large joints of the limbs (other than the knee) may be useful in arthritis of undetermined etiology especially when infection is considered. Main technical refinements include adequate selection of an optimal approach and biopsy site using new imaging methods, use of a Tru-Cut(R) needle, and placement of the cutting window tangential to the joint surface, and selection of an optimal approach and biopsy site. Detailed technique of the synovial biopsy of hip, ankle, shoulder, and elbow joints is discussed. PMID- 11387067 TI - Lumbar Chymopapain Chemonucleolysis. AB - Lumbar chemonucleolysis is an alternative treatment of the sciatic pain due to a disc herniation. It gives good results in 70 to 80% of selected patients. When a correct technique is used, the complication rate is lesser than that of the open surgery. The indications of lumbar chemonucleolysis are larger than in the past and include lateral disc herniations, large or migrated disc herniations, and patients under 18 years or over 60 years. Further studies are necessary to determine if magnetic resonance imaging helps in patients selection and improve the success rate of chemonucleolysis. PMID- 11387068 TI - Cervical Chymopapain Chemonucleolysis. AB - Cervical chemonucleolysis probably represents the treatment of choice in case of a C6-C7 radiculalgia. This technique must be performed by specialists, given the particular anatomy which is in front of the cervical disk, within the soft tissues. For the future it is necessary to delete the manufacturer's restrictions concerning chymopapain use at the cervical level. PMID- 11387069 TI - Laser Therapy in Lumbar Herniated Discs. AB - The results of experimental and clinical studies using different laser systems in intradiscal blind and endoscopic/foraminoscopic lumbar disc operations are described. We describe the evolution from blind intradiscal techniques starting in 1989 to current endoscopic/foraminoscopic procedures in 1997. Lasers are important and useful tools in these procedures. PMID- 11387071 TI - Image-Guided Selective Nerve Blocks in the Spine. AB - Selective nerve block (SNB) in the spine is a diagnostic and therapeutic imaging procedure. It consists of placing a needle under fluoroscopic guidance in the epiradicular space, which is formed by extensions of the anterior and posterior epidural membranes. Injection of nonionic contrast will confirm the correct position of the needle. This is followed by injection of a long-acting local anesthetic and/or corticosteroid. Pain reproduction followed by pain relief is recorded. This technique enables one to identify a symptomatic nerve, produce pain relief for a variable period of time, and help select surgical candidates. PMID- 11387070 TI - Image-Guided Spinal Steroid Injections. AB - This paper has been written to stimulate the radiologist to consider the various aspects of spinal steroid injections as therapeutic procedures. Special emphasis is placed on needle guidance, asepsis, and possible problems involved. It constitutes a highly practical and variously illustrated and informative guide for the choice of steroids and various spinal joint approaches. It details the following target areas: lumbar facet joints, cervical facet joints, costovertebral joints, C1-C2 joint, intervertebral discs, epidural space, and interspinous bursa. Safety is the key element of the above-mentioned techniques. The radiologist must pay special attention to the omnipresent risk of iatrogenic infection. PMID- 11387072 TI - Steroid Injection of the Appendicular Skeleton and Sacroiliac Joints. AB - Steroid joint injections may be effective in painful joints due to degenerative disease and posttraumatic changes, as well as rheumatoid arthritis and seronegative spondylarthropathy. Fluoroscopic guidance may be necessary for deep (sacroiliac, hip, and shoulder) or small (wrist, hand, and foot) joints. Detailed technique of the steroid injection is given, especially for the sacroiliac joint. PMID- 11387073 TI - Distension Arthrography in Frozen Shoulder Syndrome. AB - Distension arthrography under fluoroscopic control may be useful in frozen shoulder syndrome. The technique consists of an intra-articular injection of 2 to 3 mL of contrast material, 3 mL of 2% lidocaine, 1.5 mL of cortivazol, and distension of the joint capsule with 30 to 40 mL of refrigerated sterile saline solution. This injection is immediately followed by a physiotherapy program. The authors report their experience of distension arthography in 30 shoulders. Results were good to very good in 90% of cases at 40-day follow-up. No recurrence of a frozen shoulder syndrome was observed and no complications were encountered in this series. PMID- 11387074 TI - Percutaneous Resection of Osteoid Osteomas. AB - Osteoid osteomas in almost all locations may be treated by percutaneous resection under CT scan guidance. The authors give their own experience with a series of 30 cases. The different techniques of percutaneous resection reported in the literature are reviewed. Advantages of the percutaneous resection are compared to those of classical surgery and other percutaneous techniques such as lesion destruction with thermocoagulation, photocoagulation, and intralesional injection of alcohol. PMID- 11387075 TI - Percutaneious Radiofrequency Treatment of Osteoid Osteomas. AB - Percutaneous radiofrequency coagulation has become a highly successful alternative to surgical management of osteoid osteoma. This review discusses the reasons for intervention, and the background scientific data that supports the radiofrequency approach. The technique is described in detail, followed by a description of the results of 59 procedures. Finally, some comparisons between the radiofrequency method and other percutaneous interventions are discussed. PMID- 11387076 TI - Percutaneous Laser Photocoagulation of Osteoid Osteomas. AB - Osteoid osteoma is a painful benign bone tumor. The curative treatment of this tumor consists of complete surgical or percutaneous excision of the nidus with immediate and dramatic relief of symptoms. Interstitial laser photocoagulation (ILP) is a low-invasive percutaneous technique of thermal destruction (coagulation) of deep-seated tumors elsewhere in the body, using low-power laser energy. The aim of ILP is the local destruction of osteoid osteoma without bone weakening. Twenty-two patients with osteoid osteoma were treated with percutaneous ILP of the nidus under computed tomography guidance. The laser energy was provided by a high-power semiconductor diode laser (805 nm) with a 400 &mgr;m optical fiber. Complete pain relief was obtained in 21 patients. Percutaneous ILP of osteoid osteoma seems to be a promising, simple, precise, and minimally invasive technique as an alternative to traditional surgical and percutaneous ablations. PMID- 11387077 TI - Percutaneous Treatment of Osteoid Osteomas: Combonation of Drill Biopsy and Subsequent Ethanol Injection. AB - Osteoid osteoma is known as a benign bone-producing tumor. Histologically, it is characterized by a highly vascularized connective tissue with fibrous bone trabeculae, osteoid, osteoblasts, and numerous osteoclasts. Clinically, patients complain of pain during the night with good response to acetylsalicylic acid. Conventional radiographs show a spindle-shaped lesion with a central lucency not larger than 1 cm in size, which represents the nidus. Osteoid osteomas are most common in the diaphysis of the long bones. A successful therapy requires complete removal of the nidus, either surgically or percutaneously. Our experience is with CT-guided percutaneous drilling of the nidus with subsequent ethanol injection to sclerose remnants of the nidus. PMID- 11387078 TI - Percutaneous Vertebroplasty. AB - Percutaneous vertebroplasty (PVP) with acrylic cement [polymethylmethacrylate (PMMA)]consists of injecting PMMA into vertebral bodies weakened by osseous lesions. The aim of PVP with PMMA is to obtain an antalgic effect by consolidation in destructive lesions of the spine. There are three major indications: vertebral angiomas, osteoporotic vertebral crush syndromes, and malignant vertebral tumors. Indications in vertebral angiomas only concern patients with aggressive clinical signs (severe pain or nervous compression) and/or aggressive radiological signs. Indications in osteoporotic vertebral crush syndromes only concern patients suffering from back pain related to one or two adjacent vertebral collapses resistant to medical treatment for several weeks. Indications in malignant vertebral tumors only concern patients suffering from severe back pain related to a destruction of the vertebral body, not involving the major part of the cortical bone. Complications of PVP occur essentially in patients with vertebral metastasis. In the great majority of cases, these complications heal under medical treatment. In patients with osteoporotic vertebral crush syndromes or vertebral angiomas, the complications are represented by the increase or onset of radiculalgias (in less than 1%), which disappear after local anesthetic injection. PMID- 11387079 TI - Percutaneous Acetabular Osteoplasty. AB - The diagnostic value of ultrasound for musculoskeletal pathology is well recognized. Ultrasound offers a low-cost, nonionizing, readily available imaging technique for the evaluation of tendons, muscles, soft-tissue masses, cysts, and other fluid collections. These advantages also make ultrasound a valuable tool for guiding a variety of musculoskeletal interventions. Its real-time capabilities permit continuous monitoring of the needle position relative to the target lesion and to surrounding structures such as vessels. Ultrasound can be regarded as the OfluoroscopyO for soft tissues. Ultrasound therefore can be used to localize and characterize the lesion, monitor the needle position during the procedure, document the efficacy of the drainage or the technique, and be used for follow-up imaging. Procedures that can be performed under ultrasound guidance include aspiration of fluid for analysis, injection of medication (steroids), decompression of cyst, bursitis or joint, abscess and hematoma drainage, treatment of calcific tendinitis, biopsy, and foreign body retrieval. PMID- 11387080 TI - Intralesional Ethibloc Injections in Bone Cysts. AB - Benign bone cysts (aneurysmal and unicameral) often require treatment to prevent pathological fractures. Percutaneous injections are less invasive than surgery. We describe the different percutaneous treatments available: local injection of steroids, calcitonin, or bone marrow in unicameral bone cysts, and Ethibloc(R)(Ethicon, Ethnor Laboratories, Nevilly Sur Seine, France) injection in both unicameral or aneurysmal bone cysts. Within the tumor, histologic modifications usually occur after these injections, due to the fibrogenic properties of the agent. Therefore, we emphasize the necessity of obtaining an histologic proof before any injection, especially in aneurysmal bone cysts. PMID- 11387081 TI - Aspiration of Tendinous Calcific Deposits. AB - Needle aspiration of tendinous calcific deposits under fluoroscopic control may be a valuable tool in selected cases of painful shoulders with chronic and debilitating pain resistant to medical treatment. This article reviews the technique and results of this treatment. The aims of this procedure are (1) to evacuate a maximum amount of calcium; (2) to fragment the residual calcific deposits to facilitate its resorption during the following weeks; (3) to reduce the inflammation secondary to the presence and migration of residual calcific deposits by in situ injection of corticosteroids. Good results are obtained in 61D74% of cases. Surgical treatment should be restricted to failures of needle aspiration. PMID- 11387082 TI - Interventional Musculoskeletal Ultrasound. AB - The diagnostic value of ultrasound for musculoskeletal pathology is well recognized. Ultrasound offers a low-cost, nonionizing, readily available imaging technique for the evaluation of tendons, muscles, soft-tissue masses, cysts, and other fluid collections. These advantages also make ultrasound a valuable tool for guiding a variety of musculoskeletal interventions. Its real-time capabilities permit continuous monitoring of the needle position relative to the target lesion and to surrounding structures such as vessels. Ultrasound can be regarded as the OfluoroscopyO for soft tissues. Ultrasound therefore can be used to localize and characterize the lesion, monitor the needle position during the procedure, document the efficacy of the drainage or the technique, and be used for follow-up imaging. Procedures that can be performed under ultrasound guidance include aspiration of fluid for analysis, injection of medication (steroids), decompression of cyst, bursitis or joint, abscess and hematoma drainage, treatment of calcific tendinitis, biopsy, and foreign body retrieval. PMID- 11387083 TI - New Injectable Composites for Bone Replacementd. AB - A new class of biomaterials is emerging: injectable composites for bone replacement. They include injectable ceramics and injectable calcium phosphate hydraulic cements, such as calcium deficient hydroxyapatite cements, dahllite cements, or brushite cements. Their main advantages are their biocompatibility, resorbability, osteoconductivity, and injectability, which allows a delivery with a syringe and a needle through a percutaneous approach. Furthermore, they can be used for the controlled delivery of antibiotics or bone morphogenetic proteins. Their mechanical properties remain nonsatisfactory as compared to orthopedic cement [polymethylmetacrylate (PMMA)], but these materials still have a great potential for improvement. There is a large field of potential clinical applications for these composites, especially in implantology, bone surgery, traumatology, interventional radiology, and rheumatology. However, the biological properties of a limited number of these compounds have been studied so far and further biological evaluations, as well as rigorous clinical studies, remain to be made. PMID- 11387084 TI - Catheterization of the Medullary Cavity of Long Bones. AB - Vascular catheterization techniques were used in bones to allow access to the entire medullary cavity through a minimal percutaneous approach. The study was carried out on human and animal anatomical specimens and in living sheep. After oblique penetration at an angle of approximately 40u to the surface of the cortical bone, we attempted to pass in the bone shaft using various types of guides and catheters. Different types of instrumentation currently available were compared for ease of advancement in the shaft and quantity of bone marrow that could be aspirated. Once the metaphysis was reached, a variety of techniques were studied to penetrate the cancellous bone and to introduce a catheter. A percutaneous approach to any part of a long bone through a cortical pathway from a remote entry site is feasible in certain diagnostic or therapeutic indications, when direct approach to pathological sites in long bones is dangerous or impossible. PMID- 11387085 TI - New Trends in Interstitial Laser Photocoagulation of Bones. AB - Interstitial laser photocoagulation (ILP) was performed in experimental models prior to clinical trials to determine the feasibility of bone photocoagulation using an 805-nm diode laser and to define parameters influencing lesion size and shape. Laser energy was applied in continuous-wave mode at a power of 2 W to an ex vivo freshly ablated pig femur. Two hundred and eighty ILPs were performed followed by histologic examinations to determine the coagulation size with a freshly cleaved fiber tip compared to a precharred fiber tip. Another study was designed to determine the temperatures achieved in bone using ILP and to correlate them with histologic findings. Histologic examination has underestimated the coagulation size with precharred fibers that varied from 3.4 mm (200 J) to 9.2 mm (1200 J) in diameter. Thermal data were significantly different with presumed lesions of 16 mm in diameter for 1200 J energy. PMID- 11387086 TI - New Trends in Musculoskeletal Interventional Radiology: Percutaneous, MR_Guided Skeletal Biopsy. AB - Several different types of magnetic resonance (MR) scanners that allow access to the patient during image acquisition and thus permit MR-guided biopsies have been introduced. A variety of MR-compatible biopsy needles and a prototype coaxial drill system, powered either by hand or an optional motor, are presently available for sampling skeletal lesions. In a small study population of 28 patients in our hospital who required a biopsy of a skeletal lesion, all but five biopsy procedures could be completed within the MR unit. In one patient who required a transpedicular approach to a lumbar vertebra, we decided to switch to computed tomography (CT) guidance. Four patients needed general anesthesia, which is currently not available in our interventional MR suite. Those procedures were also performed under CT guidance. In 18 of the 23 MR-guided cases, the samples were sufficient and the histopathologic diagnoses were confirmed by surgery or the clinical course. Three biopsies contained an adequate volume in the sample, but the contents were either unrepresentative of the final pathology or distorted beyond recognition by the biopsy drill. Two others revealed insufficient material. No procedural complications occurred. Percutaneous biopsy of skeletal lesions performed under MR guidance seems to be a safe procedure and reasonably accurate given the fact that it is still in an early stage of development. MR imaging may be used as an alternative to CT, but its role vis-vis CT has yet to be ascertained. PMID- 11387087 TI - Value of Percutaneous Treatments of the Lumbar Spine in Back and Nerve Root Pain. AB - Percutaneous treatments are widely used in low back pain (LBP) and nerve root pain. The only treatment assessed in several controlled studies is chemonucleolysis. Automated percutaneous discectomy was proven non-effective in two controlled studies and manual percutaneous discectomies still require appropriate evaluation. The problem of facet joint treatments is to select patients with LBP originating from these joints. Epidural injections of steroids are the cornerstone of the medical management in sciatica, but the data from controlled studies are inconsistent. Further studies are needed to give guidelines on the best time after the onset of radicular pain, volume, number, and timing of the injections. In addition, the problem of misplacement of the needle should be considered. Foraminal injections of steroids under fluoroscopic control also need controlled studies. Endoscopic procedures in the epidural space should be studied by leading teams representing the spine specialists in the community. Finally, in addition to their technical aspect, each new percutaneous procedure should be evaluated by a stringent methodology to assess the risk benefit. PMID- 11387088 TI - Cervical Vertebral Trauma: Plain Film Approach. AB - The plain film assessment of cervical vertebral trauma remains a cornerstone of emergency radiology. An understanding of proper film technique, as well as a familiarity with the significant signs of cervical column injury will enable the radiologist to interpret the radiograph accurately. If further imaging studies are warranted, accurate interpretation of the plain films will insure that the proper study is obtained. PMID- 11387089 TI - Helical CT for the Evaluation of Cervical Vertebral Injuries. AB - The imaging evaluation of the cervical vertebral column in patients subjected to nontrivial trauma has been a subject of major debate in trauma care. A variety of protocols have been proposed that comprise different combinations of radiographs with or without computed tomography (CT) and magnetic resonance imaging in selected cases. In our institution, helical computed tomography (HCT) of the cervical region has been used in the initial assessment of trauma patients either as a screening modality or as an adjunct to radiography. Our results support the use of HCT because it has improved fracture detection and provided better depiction of the extent of injury. More importantly, we have found that significant fractures can be missed when exclusively relying on radiography. In addition, the time for clearance of the cervical column and the disposition time of patients from the trauma area have been reduced while improving the utilization of CT. PMID- 11387090 TI - MR Imaging for Assessing Acute Vertebral Trauma. AB - Magnetic resonance imaging (MRI) has revolutionized the imaging assessment of patients sustaining acute vertebral injury and is indicated for all hemodynamically stable patients with acute neurologic deficits related to spinal column injury, particularly in the cervical region. MRI defines the presence and extent of lesions involving osseous structures, ligaments and other soft tissues, and the spinal cord parenchyma. Information obtained from MRI is useful in assessing the indication for and best approach to surgical management of vertebral injury by revealing herniated disc material, epidural hematoma, significant osteophytes, and level(s) of probable or potential spinal column instability. The appearance of spinal cord lesions by MRI provides prognostic information regarding likely extent of recovery of neurologic function. Magnetic resonance angiography (MRA) can reliably demonstrate vertebral artery injuries not uncommonly associated with cervical spine subluxation/dislocation and fractures crossing the foramen transversarium. Improvements in speed of MR image acquisition and patient physiological support and monitoring compatibility in the MR-environment is making MRI more available and safe for use in the setting of acute major trauma. PMID- 11387091 TI - Thoracic and Lumbar Vertebral Injuries. AB - Injuries to the thoracic and lumbar vertebral column are not uncommon and are the result of increased loading on structures endowed with little motion or flexibility. Conventional radiographs, when carefully analyzed for the OfootprintsO and OfingerprintsO of the injury, will define its nature and, to a degree, its extent. A system of radiologic analysis is described and the parameters for diagnosis and differentiation are given. CT and MR imaging will provide further information regarding the full extent of injury and involvement of the spinal cord, if any. The importance of the radiographic signs described in the text is discussed in regard to posttraumatic stability of the vertebral column. PMID- 11387092 TI - Imaging of Facial Trauma. AB - Severe injury to the facial skeleton is a frequent finding in victims of trauma. Severe facial fractures produce a variety of radiographic and CT abnormalities that are not difficult to recognize if one carefully analyzes the images. This discussion covers a number of radiographic findings that are characteristic of several families of major injuries to the midface. The following types of injuries are discussed: orbital, zygomaticomaxillary complex, and LeFort. PMID- 11387093 TI - Pelvic Injuries. AB - Pelvic injuries are an important consideration in trauma patients. This is largely as a result of the major disruptive forces required to fracture the pelvic ring, and the resulting complications that can be expected from major pelvic ring disruption. These can be both immediate (associated visceral injury, hemorrhage, etc.) or delayed (infection, morbidity, etc.), but to a large extent can be anticipated by knowledge of the type of injury force producing the fracture, and specific complications associated with each force of injury. This article will examine pelvic injuries from a mechanistic approach, determining the specific pelvic fracture patterns that are encountered, how they may be imaged best, and what types of complication might be expected. PMID- 11387094 TI - CT Evaluation of Laryngotracheal Trauma. AB - Laryngotracheal injuries are rare, and typically associated with multisystem trauma. They may be blunt or penetrating in nature, and are in the great majority of cases related to motor vehicle accidents or OclotheslineO injuries with a small percentage due to direct blows sustained during assaults or athletic contests, hanging or manual strangulation, or other less common etiologies including iatrogenic causes. Missed diagnoses or mismanagement may result in the patient's death or significant long-term morbidity. The radiologist must be familiar with the normal computed tomographic (CT) appearance of laryngotracheal anatomy to correctly interpret CT studies following injury, and must also be aware of the central role that CT plays in diagnosis, management, and selection of therapy. This should include an understanding of the Shaefer classification of laryngeal injuries that is based on a combination of the CT and endoscopic findings. Although an acceptable evaluation of the traumatized larynx is obtainable with most commercially available CT scanners, optimal studies are produced by CT devices capable of spiral technique and subsecond scan times, particularly in regard to their ability to generate thin retrospectively reconstructed two-dimensional (2D) axial sections, 2D coronal and sagittal images, and three-dimensional (3D) images. Our discussion of laryngotracheal injuries is divided into four parts. Part 1 deals with injuries to the endolaryngeal soft tissues structures, including the mucosa, vocal cords, and deep compartments. The ability of CT to demonstrate endolaryngeal edema and hematoma, vocal cord injuries, subcutaneous emphysema, and aspirated radiopaque foreign bodies is discussed along with its inability to demonstrate the site of mucosal perforations or degloving injuries. Part II deals with fractures of the hyoid bone, epiglottis, and thyroid and cricoid cartilages, while Part III discusses dislocations of the cricoarytenoid and cricothyroid joints. Finally, Part IV discusses laryngotracheal separation, the most immediately life threatening laryngotracheal injury, and the difficulties inherent in making this diagnosis prospectively by CT. PMID- 11387095 TI - MR Imaging of the Elbow: Technical Considerations. PMID- 11387096 TI - Normal Anatomy of the Elbow on Conventional MR Imaging and MR Arthrography. AB - Conventional MR imaging allows clear depiction of the muscles, tendons, nerves, vessels, ligaments, bones, and cartilage that compose the elbow. MR arthrography can be a valuable supplementary technique for optimizing evaluation of intraarticular structures, including the undersurfaces of the collateral ligaments. Regardless of the imaging technique utilized, knowledge of normal anatomy-and normal anatomic variants-is fundamental for accurate assessment of normal and diseased states. PMID- 11387097 TI - MRI of Normal Variants and Interpretation Pitfalls of the Elbow. AB - One of the fundamental principles of MR imaging interpretation is the ability to distinguish normal anatomical landmarks from true disease. The radiologist is thus compelled to accumulate a comprehensive knowledge of normal structures, variants, and potential MRI diagnostic pitfalls.1-5 In this article we will focus on a number of normal bony, ligamentous, and tendinous structures that can simulate disease at the elbow. We discuss the particular anatomy responsible for the appearance of each of these interpretation pitfalls as well as the distinguishing features between these normal variants and true disease. PMID- 11387098 TI - Collateral Ligaments. AB - The collateral ligaments of the elbow are responsible for stability in response to varus and valgus stress. Injury to the collateral ligaments can be acute, but it is usually due to repetitive microtrauma. In the general population, collateral ligament injury is rare, but in the appropriate individual with the proper forces applied to the elbow, this type of abnormality is not uncommon. The elbow is the second most commonly injured joint because of overuse. Magnetic resonance imaging (MRI) is the best technique available for visualization of the ligaments. Contrast injection further enhances identification of these structures and helps to define pathological conditions affecting them. This article demonstrates some normal anatomy and shows examples of collateral ligament pathology in unenhanced and enhanced MR images. PMID- 11387099 TI - Tendon Disorders. AB - The clinically important tendons around the elbow include the biceps and triceps brachii and the flexor and extensor muscles of the forearm. Familiarity with the normal magnetic resonance (MR) appearance of these structures facilitates the MR diagnosis of tendon abnormalities. Often referred to as epicondylitis, degeneration of the flexor or extensor groups is a common clinical complaint, treated conservatively and usually not requiring MR imaging. Imaging may play a role in unusual or severe cases; elevated signal in the appropriate common tendon origin is typically seen on T2 weighted images. Significant injury or disruption of the distal biceps or triceps tendons is a rare event, usually related to an acute event. Discontinuity of these tendons is best imaged using sagittal and axial T2 weighted sequences. Partial tears may demonstrate tendon irregularity and elevated signal within and around the tendon. PMID- 11387100 TI - Nerve Entrapment. AB - Entrapment and compressive neuropathies are frequent clinical conditions occurring about the elbow. In most instances clinical and electromyograhic evaluation are adequate for patient management, but in some cases further evaluation with imaging techniques is required. Magnetic resonance imaging (MRI) has been shown to be useful in the evaluation of these conditions, especially to detect space occupying lesions. In this article, compressive neuropathies involving the ulnar, median, and radial nerves are discussed, with emphasis on the normal anatomy and the MRI depiction of pathologic findings. PMID- 11387101 TI - MR Imaging of Pediatric Elbow Trauma. AB - Magnetic resonance imaging (MRI) of the pediatric elbow is important because it allows delineation of fracture type in a joint with complex anatomy, made more complex by the variable appearance of ossification centers of the distal humerus.1-4 Acute and chronic injuries can be evaluated with MRI using several classification systems. Accurate diagnosis of pediatric elbow trauma with MRI has management implications. For example, a Salter-Harris II fracture can be differentiated from a Salter-Harris IV physeal fracture by delineating intra articular fracture extension and displacement, which requires open reduction internal fixation. The former is treated with closed reduction if nondisplaced, and the latter, with open reduction and internal fixation. The rate of complications in this age group is high and the treatment for these deformities thereafter can be difficult. There is also a useful role for MRI when the clinical history is unobtainable or absent as in a case of child abuse, for example. The multiplanar capability of MR allows for good preoperative planning to delineate fragment displacement. The main fracture classifications are presented. PMID- 11387102 TI - Musculoskeletal Ultrasound: Basic Principles. AB - Equipment selection and examination technique are critical issues in the practice of musculoskeletal ultrasound. PMID- 11387103 TI - Sonography of the Shoulder. AB - A great amount of work in musculoskeletal sonography has focused on evaluation of the shoulder and rotator cuff. This is primarily due to the fact that the shoulder is a common site of symptomatology and clinical evaluation is difficult. Even though sonography of the rotator cuff is more difficult than other large tendons, there has been constant incentive to develop and refine shoulder sonography. In the past five years there have been dramatic improvements in high resolution transducers, as well as advances in our understanding of the technique of shoulder sonography and more widespread agreement of the findings seen with rotator cuff tears. All of these factors have contributed to making the exam easier to perform and interpret than in the past. Ultrasound has now evolved into a mature modality for evaluating rotator cuff tears with expected sensitivities of better than 90%. PMID- 11387104 TI - Ultrasound of the Elbow. AB - Ultrasound is ideally suited for the evaluation of soft tissue lesions of the elbow. The ultrasound study of the elbow is rapid, dynamic and cost-effective. This article presents a brief overview of scanning technique and illustrates some common pathologies. PMID- 11387105 TI - Sonography of the Wrist and Hand. AB - With the advent of new technical improvements and innovations, sonography is gaining ground in use and acceptance as a highly useful modality to diagnose a variety of diseases involving the wrist and the hand. Common ailments such as tendinitis and tenosynovitis can be diagnosed easily, and the diagnosis of small soft tissue masses such as ganglia has become almost routine. Sonography holds great promise in the evaluation of disease involving the median and the ulnar nerves, the ligaments of the wrist, and the triangular fibrocartilage. It is the ideal modality in the diagnosis of intra-articular loose bodies. PMID- 11387106 TI - Ultrasonography of the Knee. AB - Musculoskeletal ultrasonography (MSUS) is a radiologic modality that can help in the evaluation of the knee and its lesions. Both soft tissue and surface bony structures can be sonographically visualized. The extra-articular structures of the knee can be evaluated, specifically the ligaments, bursae, and tendons. Collateral ligaments and the extensor mechanism, especially quadriceps and patellar tendons, are readily visualized. The diagnosis of a Baker's cyst and its complications is a common indication. The intra-articular structures of the knee, with proper maneuvers and transducers, can also be evaluated. The femoral condylar and patella cartilage can be examined for traumatic or degenerative lesions. The anterior and posterior cruciate ligaments can be partially visualized directly, or evaluated indirectly. Loose bodies are readily detectable whether intra-articular or extraneous in a Baker's cyst. Synovial disease can be characterized without the use of radiographic contrast. MSUS can be used as a screening, diagnostic or follow-up radiologic tool in the diseases of the knee. PMID- 11387107 TI - Ultrasound of the Foot and Ankle. AB - For specific indications, ultrasound is an inexpensive and efficient alternative to magnetic resonance (MR) imaging of the ankle and foot. In addition to the tendons, the anterior joint, retrocalcaneal bursa, ligaments, plantar fascia, and soft tissues can be imaged with ultrasound. Tenosynovitis, tendon tears and tendinosis, joint effusions, intra-articular loose bodies, ganglion cysts, plantar fasciitis, and Morton neuromas can be demonstrated with ultrasound. In most cases, a focused ankle or foot ultrasound can be performed more rapidly and efficiently than magnetic resonance imaging. Direct correlation with the site of pain and rapid comparison with the asymptomatic ankle are possible with this modality. As pressure for cost containment continues, demand for ultrasound of the ankle and foot will likely increase. PMID- 11387108 TI - Ultrasound of Soft Tissue Masses. AB - Ultrasound (US) should be considered an integral part of the imaging arsenal of the musculoskeletal radiologist. It offers many advantages over other imaging modalities of which, the contact between the radiologist and the patient at the time of examination and its dynamic imaging capabilities are unique. Coupled with color Doppler it gives important information on the vascularity of soft-tissue masses, and it provides rapid, real-time guidance for fine-needle aspiration and biopsy. This article discusses the role of US in the diagnosis of soft-tissue masses. We also propose an algorithm for the investigation of a patient with a suspected soft-tissue mass which uses US and Doppler studies as the second-line imaging modality after radiographs. US is helpful in the diagnosis of cystic and solid soft-tissue masses but, as with any other imaging studies, it is important to know its limitations. Sometimes MRI should be done in place of, or to complement the US examination. In many cases, a biopsy is needed to arrive at a precise and final diagnosis. PMID- 11387109 TI - Ultrasound of Musculoskeletal Infections. AB - Ultrasound (US) can play an important role in differentiating infectious processes from other musculoskeletal diseases. US is the imaging modality of choice to diagnose superficial abscesses. These abscesses may have variable echogenicity and their detection can be facilitated by ultrasonographic dynamic evaluation and color Doppler imaging. Septic arthritis either in a native or prosthetic joint may have devastating complications on the joint function. Early diagnosis requires joint fluid analysis. US is very sensitive in the detection of joint effusions and is helpful to guide the arthrocentesis. US may help in the early diagnosis of osteomyelitis, although one must always remember that a normal US never excludes bone infection. This article also reviews the role of US in the diagnosis of necrotizing fasciitis, tenosynovitis, bursitis, and pyomyositis. After initial radiographs, along with nuclear medicine studies, US should be considered as the next imaging step in the investigation of musculoskeletal infections. PMID- 11387110 TI - Ultrasound in Arthritis and Inflammation. AB - High resolution ultrasound (US) provides a versatile and sensitive method of demonstrating both soft-tissue and osseous changes in early arthritis and other inflammatory conditions. A brief outline is provided regarding the potential usage of US in the management of patients with musculoskeletal inflammatory disease. PMID- 11387111 TI - Musculoskeletal Ultrasound in Pediatrics. AB - Ultrasound is ideally suited to the evaluation of the pediatric musculoskeletal system because of the increased ratio of cartilage to bone in the immature skeleton. The purpose of this article is to review the current uses of musculoskeletal ultrasound in pediatric patients. Hip sonography is widely accepted; other applications are increasing in popularity. PMID- 11387112 TI - Power Doppler Sonography: Applications in Musculoskeletal Imaging. AB - Power Doppler sonography (PDS) is a recently developed technique that depicts moving blood volume within tissues. The enhanced sensitivity of PDS makes it possible to depict soft tissue vascularity, facilitating its role in the evaluation of musculoskeletal inflammatory diseases. A broad range of new and potential applications of PDS in the musculoskeletal system will be described including its use in the setting of bursal, tendon, articular, and muscle diseases, and in the evaluation of fluid collections and soft tissue masses. PMID- 11387113 TI - Complications of Arthrography. AB - Serious reactions to intra-articular contrast media are exceedingly rare, and arthrography is considered to be a very safe procedure. To assess the safety of arthrography, a questionnaire was sent to 180 radiologists experienced in arthrography. The 134 respondents had performed approximately 262,000 arthrograms throughout their career. Their experience included 45 cases of infection, eight cases of anaphylaxis, and five cases of vascular complications; no deaths were reported. Most of the reactions reported were minor and included pain, vasovagal reactions, and urticaria. In addition, the respondents reported approximately 13,300 magnetic resonance (MR) arthrograms, which had been performed with intra articular gadolinium. Of these, there were only six cases of reported reactions, all of which were minor, including vasovagal reactions, pain, and headaches. PMID- 11387114 TI - MR Arthrography of the Hip. AB - Magnetic resonance imaging (MRI) of the hip has been valuable in the diagnosis of occult osseous abnormalities and of periarticular soft tissue disorders. MRI has been less useful in the evaluation of acetabular labral tears and other intra articular abnormalities. Image optimization is more technically challenging in the hip than in smaller joints because the overlying soft tissues are thicker, resulting in decreased spatial and contrast resolutions that may not be adequate to distinguish the acetabular labrum from subchondral bone, articular cartilage, and joint capsule. MR arthrography (MRA) extends the capabilities of conventional MRI because contrast solution separates intra-articular structures and outlines abnormalities. In hips with suspected acetabular labral injury, arthrographic MR images demonstrate the location and length of tears and the presence of associated capsular defects. Arthrographic MR images may also enable the accurate diagnosis of cartilage lesions and intra-articular loose bodies. This article illustrates normal arthrographic MR features of the hip as well as pathologic disorders of the acetabular labrum, capsule, and articular cartilage PMID- 11387115 TI - CT and MR Arthrography of the Glenoid Labroligamentous Complex. AB - Because they distend the shoulder joint and bathe the labrum with contrast, CT arthrography (CTa) and MR arthrography (MRA) are both excellent tests for evaluating the labrum. In this article, we discuss the advantages and disadvantages of each examination. We also describe techniques for performing CTa and MRA, including how to maximize image quality. The normal anatomy of the labrum and glenohumeral ligaments is described and demonstrated on both CTa and MRA images. To improve communication with orthopedic surgeons, the discussion of the pathology and normal variants of the labroligamentous complex is organized by functional anatomic regions. CTa and MRA images are then used to demonstrate a variety of pathologic and normal variant appearances of the labroligamentous complex. PMID- 11387116 TI - Gadolinium MR Arthrography of the Rotator Cuff. AB - Conventional magnetic resonance imaging (MRI) of the rotator cuff has not done well in distinguishing partial-thickness tears from tendonitis or small, full thickness tears. However, these are important distinctions for orthopedic surgeons who are deciding whether to operate and what type of surgery to perform on a patient with suspect rotator cuff pathology. Gadolinium magnetic resonance arthrography (MRA) involves injecting dilute gadolinium into the shoulder under fluoroscopy. Subsequent multiplanar T1-weighted fat-suppressed fast spin echo combined with T2-weighted fast spin echo and short tau inversion recovery (STIR) produces images that can accurately differentiate absence of tear from partial thickness tear and from small, full-thickness tear; as a fringe benefit, this combination of sequences can identify the inflammation associated with tendonitis. Furthermore, the technique differentiates articular from bursal surface partial-thickness tears, and it accurately quantitates the size of full thickness tears to help surgeons choose between arthroscopic versus open surgery. PMID- 11387117 TI - MR Arthrography of the Knee. AB - The role of magnetic resonance imaging (MRI) in the evaluation of musculoskeletal injuries has been well documented during the last decade. There remain several important clinical situations in which noncontrast MRI has been disappointing. In the knee, magnetic resonance arthrography (MRA) can supplement noncontrast MRI in the evaluation of specific conditions, such as postoperative meniscus and osteochondritis dissecans. MRA significantly increases accuracy in the diagnosis of meniscal retear, as is seen in cases in which there has been a meniscal resection of more than 25% or after meniscal suturing. Also, in the evaluation of osteochondritis dissecans, the addition of intra-articular contrast has proved beneficial. Aside from assessing the integrity of the articular cartilage surface and documenting loose bodies, the contrast allows the clinician to distinguish fibrovascular granulation tissue from fluid partially or completely surrounding the osteochondritic fragment. These topics are discussed in this article. PMID- 11387118 TI - MR Arthrography of the Elbow and Wrist. AB - This article describes the present status and future directions of magnetic resonance arthrography (MRA) of the elbow and the wrist. The indications are evolving and not yet clearly defined. MRA of the elbow is useful for evaluation of collateral ligaments, the articular surfaces, and suspected loose bodies. MRA of the wrist currently is typically reserved for difficult cases. A description and a comparison of direct and indirect MRA techniques is provided. Direct MRA produces reliable joint enhancement with capsular distension. Indirect MRA does not require a radiologist experienced with arthrography but if it is improperly performed, results in suboptimal delineation of intra-articular structures. A review of pertinent normal anatomy and common variants is included as well as a discussion of pathology that is suitably demonstrated by MRA technique. Typical examples of normal variants, abnormalities, and pitfalls are illustrated to reduce interpretive error for radiologists using MRA of the elbow and the wrist. PMID- 11387119 TI - Anesthetic and Corticosteroid Joint Injections: A Primer. AB - Complex joint anatomy may render the clinical diagnosis of a patient with joint pain difficult. Pain may be referred to a joint from an adjacent area (e.g., from the back to the hip) making the diagnosis difficult. The radiologist with the use of fluoroscopy is the ideal person to perform diagnostic and therapeutic joint injections. Long-acting anesthetic alone or combined with a corticosteroid may help the clinician localize the cause of the joint pain and subsequently institute the proper therapy. This article includes a discussion of the commonly used injectable corticosteroids that are available. The choice of corticosteroid is based on personal preference. Depomedrol (Upjohn, Kalamazoo, MI) is the least expensive corticosteroid available; however, a relatively insoluble drug such as Aristospan (Lederle, Deerfield, IL) or a combination drug such as Celestone Soluspan (Schering, Kenilworth, NJ) may be a better choice in rheumatoid arthritis. The technique used to inject the hip and the shoulder is discussed. PMID- 11387120 TI - Diagnostic and Therapeutic Foot and Ankle Injections. AB - For patients with disabling foot or ankle pain, medical or surgical treatment decisions can be difficult to make when multiple joints show changes of osteoarthritis or if the patient's pain clinically is related to a joint or tendon that is normal by other imaging studies. For these patients, injection of anesthetic, steroid, or both, into joints or tendon sheaths of the foot and ankle provides important diagnostic information and therapeutic relief. Diagnostic injections may show that the joints noted by other imaging studies have osteoarthritis that are not responsible for a patient's pain or that a normal joint is responsible. When multiple joints show changes of arthritis, anesthetic injections can help decide which and how many joints could benefit from surgical arthrodesis. Relief of pain after anesthetic joint injection correlates well with postoperative pain relief subsequent to arthrodesis. This article discusses the indications and the contraindications for performing diagnostic and therapeutic joint injections, and also presents the techniques for performing these studies. PMID- 11387121 TI - Subtalar Joint Arthrography. AB - The purpose of this study was to determine the accuracy and the specificity of an optimum technique of posterior talocalcaneal/posterior subtalar (PST) joint arthrography and anesthetic injection in patients with hindfoot pain. Fifty-five PST joints were studied in 55 patients. The posteromedial approach was used in the first 24 patients, followed by an anterolateral approach in 31 patients. The ease of performance, success of confirming PST needle position, and adverse effects were noted. After contrast injection, a combination of 1% lidocaine and 0.5% bupivacaine was injected. Results consisted of 47 arthrographically confirmed PST injections. The posteromedial approach was deemed more difficult; three patients had tendon sheath opacification and four had unwanted anesthesia of the toes. The anterolateral approach was technically easier and no extra articular structures were visualized or anesthetized. Therefore, PST arthrography with anesthetic injection is optimized with an anterolateral approach. PMID- 11387122 TI - Arthrosonography. AB - Ultrasound plays an important role in the evaluation and management of joint disease. In this article, applications of sonography in the diagnosis of articular disease are detailed. General arthrosonographic technique is reviewed, emphasizing normal anatomy and pathologic conditions, and applications at specific joints are discussed. Finally, ultrasound-guided joint interventions are described, including indications and technique. PMID- 11387124 TI - MR Imaging of Soft Tissue Tumors: An Overview. AB - Over the past decade, magnetic resonance (MR) imaging has been established as the most widely used examination for the detection of a suspected soft tissue mass and often for depiction of the extent and the characterization of the mass. Sensitivity of the examination for the detection of soft tissue masses is high but specificity is limited. With experience accrued over the past decade, radiologists are increasingly able to predict the histology of lesions based on MR imaging criteria. However, because of the overlap of morphologic features of benign and malignant soft tissue masses, a benign-appearing mass should not be considered benign unless the tumor can be specifically named and its histology predicted by well-established MR imaging features. When a mass is not thought to be an overt sarcoma and there is uncertainty as to whether it is benign or malignant, the tumor should be characterized as indeterminate and approached as a sarcoma until proven otherwise. This article reviews MR imaging in diagnosing and staging soft tissue masses and briefly discusses the radiologistOs approach to percutaneous biopsy of indeterminate soft tissue masses. PMID- 11387125 TI - Imaging of Soft Tissue Neoplasms in the Adult: Benign Tumors. AB - This discussion reviews the spectrum of benign soft tissue tumors usually found in adults. Lesions most commonly identified in the pediatric population, tumorlike masses, and malignant tumors are discussed in other articles in this issue. Rather than presenting a complete review, the focus of this article is on benign tumors in which the diagnosis may be confidently made or strongly suggested on the basis of imaging. Diagnoses presented include those for lipoma, lipoma variants, giant cell tumor of the tendon sheath, pigmented villonodular synovitis, superficial and deep fibromatoses, elastofibroma, and peripheral nerve sheath tumors. PMID- 11387123 TI - Clinical Evaluation and Treatment of Soft Tissue Tumors. AB - Soft tissue sarcomas are uncommon and frequently missed on examination, resulting in delays in diagnosis and, occasionally, inappropriate treatment. Sarcoma staging, the process of defining the local extent of tumor and potential distant spread, involves a thorough history and physical examination, directed imaging, and biopsy. Biopsy is a complicated procedure in approximately 20% of cases and should be performed only by experienced personnel and at a center with a multidisciplinary team familiar with the treatment of patients with soft tissue sarcomas. The goal of surgery is to obtain tumor-free margins. In conjunction with radiation therapy, surgery can then provide local disease control in more than 90% of patients. The role of chemotherapy in nonmetastatic disease is unclear and is of marginal efficacy in patients with metastases. Although most tumors recur within 2 to 5 years, long-term clinical and radiographic surveillance is necessary. PMID- 11387126 TI - Imaging of Soft Tissue Neoplasms in the Adult: Malignant Tumors. AB - This review discusses the spectrum of malignant soft tissue tumors usually found in adults. Lesions most commonly identified in the pediatric population, as well as tumorlike masses, are not covered. Rather than presenting a complete review, the focus of this discussion is on common lesions and those lesions in which the diagnosis may be suggested on the basis of imaging. Diagnoses covered include malignant fibrous histiocytoma, fibrosarcoma, dermatofibrosarcoma protuberans, liposarcoma, synovial sarcoma, malignant peripheral nerve sheath tumor, leiomyosarcoma, aggressive vascular tumors, extraskeletal chondrosarcoma and osteosarcoma, lymphoma, and metastatic carcinoma. PMID- 11387127 TI - Imaging of Soft Tissue Tumors in the Pediatric Patient. AB - The presence of a soft tissue mass in children is of concern to parents and physicians. Fortunately, these masses are rare and usually benign or pseudotumoral. When dealing with malignant soft tissue tumors, therapeutic options and long-term survival are strongly related to the disease stage at the time of diagnosis. Therefore, when children present with indeterminate or persisting symptoms and posttraumatic, metabolic, or infectious disorders have been ruled out, one should perform dedicated imaging studies (conventional radiography, computed tomography [CT], or both; sonography; magnetic resonance [MR] imaging) to exclude the possibility of a nonpalpable soft tissue mass or to characterize the mass when present. An overview of the use of the different imaging modalities for evaluating soft tissue tumors in the pediatric patient is presented. Because of the numerous benign, malignant, and pseudotumoral soft tissue masses that are often encountered in children, clinical, histologic, and imaging features are presented as concise tables. PMID- 11387128 TI - The Radiologic Features of Non-Neoplastic Tumors of Soft Tissue. AB - Neoplasms of the soft tissues cause localized swelling and a variable degree of tissue response on the part of the host; these features they share with many non neoplastic disorders. A spectrum of lesions that may simulate soft tissue neoplasms are described, with their radiologic appearances. The cellular nature of the matrix of a lesion cannot be identified absolutely as neoplastic by current imaging methods. Although sonography and magnetic resonance imaging can each produce valuable diagnostic information hitherto not provided by imaging the soft tissues, they do not per se show evidence of neoplasia. The differentiation of the two types of tumor, neoplastic and non-neoplastic, can only be achieved by a combination of clinical, radiologic and histologic information. Ultimately, biopsy with histologic examination may be required for the definitive diagnosis. PMID- 11387129 TI - Dynamic Contrast-Enhanced MR Imaging for Soft Tissue Sarcomas. AB - Dynamic contrast-enhanced magnetic resonance imaging has recently emerged as an important method for evaluating soft tissue sarcomas for biopsy localization, chemotherapeutic response, and long-term follow-up because of its ability to detect viable tumor. This article presents the basic principles of contrast kinetics in soft tissue sarcomas after bolus injection of contrast agent and discusses the current postprocessing methods (subtraction, first-pass image and time-intensity curves with region of interest, and color-encoded techniques) used to display these dynamic studies. Because of its excellent temporal resolution, dynamic MR imaging can delineate the early uptake of contrast agent in sarcomas within seconds after injection, almost synchronous with arterial enhancement, and thereby differentiate the rapidly enhancing viable tumor from the nonenhancing necrotic tumor and the late enhancing changes after surgery, radiation therapy, and chemotherapy. PMID- 11387130 TI - Soft Tissue Masses: The Underutilization of Sonography. AB - For the evaluation of soft tissue masses, sonography (US) has proved to be very accurate in confirming the presence or absence of a lesion, with a very high negative predictive value. Many soft tissue masses result from trauma, inflammation, infection, or cystic changes and are not true neoplasms. In the proper clinical context, US can diagnose a muscular tear, hernia, myositis ossificans, or rhabdomyolysis. Retained foreign bodies are readily identified and localized with US. US differentiates between cellulitis and abscess and can diagnose masses resulting from tendinitis, tenosynovitis, or bursitis. It is the modality of choice for diagnosing cysts, including intact or ruptured BakerOs cysts and ganglion cysts in the distal extremities. Among benign neoplasms, lipomas and hemangiomas display a wide spectrum of echogenicity. US can diagnose nerve sheath tumors by demonstrating the connection between the mass and the normal nerve. Except for some well-differentiated liposarcomas, which may appear echogenic, the vast majority of malignant tumors in the soft tissues are hypoechoic. Real-time US is ideal for guiding large-core needle biopsy of soft tissue sarcomas. US is extremely sensitive in detecting early recurrences after surgical excision, which are readily confirmed by US-guided fine-needle aspiration. Any nonpalpable mass visualized by US can be conveniently localized pre- or intraoperatively with US guidance. Provided the examination is done by a well-trained operator using state-of-the-art equipment, the cost-effectiveness of US justifies its use as a first-line examination technique in many situations involving soft tissues, with magnetic resonance imaging being the problem-solving tool and staging procedure. PMID- 11387131 TI - Positron Emission Tomography in Grading Soft Tissue Sarcomas. AB - Soft tissue sarcomas present with varied radiological appearances. Positron imaging with [F-18] fluorodeoxyglucose (FDG PET) has recently made promising contributions to management of patients by providing a noninvasive means for evaluating tumor metabolism and providing important biological information about soft tissue malignant tumors. PET imaging not only gives quantitative data on metabolic rates of tumors but can also readily provide semiquantitative data of uptake of tumors by measuring uptake ratios. These values have been helpful for noninvasively grading tumors. This value is called the tumor standard uptake value (SUV). The tumor grades (low, intermediate, high) mean SUV values show a high level of significance in discrimination among tumor grade groups. PMID- 11387132 TI - Percutaneous Biopsy of Primary Soft Tissue Tumors. AB - This article reviews the technique, results, and complications of percutaneous biopsies of soft tissue tumors. Biopsy of soft tissue tumors may be achieved either with fine needles or core-biopsy needles. The choice between the two biopsy techniques ultimately depends on the experience of the radiologist and pathologist. Since an inadequate technique may severely affect final treatment, the biopsy track should be discussed with the surgeon responsible for future management each time a primary tumor is suspected. PMID- 11387133 TI - The Role of Immunohistochemistry in the Diagnosis of Soft Tissue Tumors. AB - Soft tissue tumors may be the most diagnostically challenging lesions in pathology. Although clinical features and morphologic pattern are the gold standard for the diagnosis of soft tissue lesions, immunohistochemistry can confirm the phenotype, or resembled cell of origin, and sometimes the biologic potential of the lesion. This article discusses the role of immunohistochemistry in the diagnosis of soft tissue tumors. PMID- 11387134 TI - Basic Concepts in Molecular Cytogenetics of Soft Tissue Tumors for the Clinician. AB - Over the past several years, cytogenetic and molecular analyses have played a growing adjunct role in the clinicopathological evaluation of soft tissue tumors. Recent technological advances, especially in fluorescence in situ hybridization and polymerase chain reaction, have enabled the analysis of frozen and paraffin embedded tissue as well as fresh tumor samples. Many characteristic genetic abnormalities have been identified that are of diagnostic utility in the analysis of soft tissue tumors. Additionally, certain genetic aberrations have been found to be of potential prognostic value. With the abundance of useful tools that are available, molecular cytogenetic analyses are likely to become an integral part of the analysis of soft tissue tumors. These analyses can be performed readily using small amounts of tumor (e.g., from sonographically or computed tomographic guided percutaneous biopsy specimens). PMID- 11387135 TI - Bone Metastases from Soft Tissue Sarcomas. AB - The incidence, distribution, time of appearance, and radiologic findings of bone metastases from soft tissue sarcomas, exclusive of lymphomas, were evaluated in 320 patients with soft tissue sarcomas. Thirty patients (9.4%) had evidence of 58 bone metastases. Five of 30 patients presented with metastases, and 25 of 30 patients developed metastases up to 66 months after presentation with a mean time interval of 21.3 months. The incidence of skeletal metastases differed among histologic subtypes of sarcomas; alveolar soft part sarcoma (5 of 8), dedifferentiated liposarcoma (2 of 4), angiosarcoma (2 of 4), and rhabdomyosarcoma (5 of 16) tended to show a higher incidence of bone metastases. The sarcomas metastasized to the regional bones close to the primary tumor in 16 (53%) of 30 patients and to the axial bones in 18 (60%). On conventional radiographs, the osseous metastases demonstrated predominantly osteolytic changes, and evidence of pathological fracture was observed in 31% of 58 metastases. PMID- 11387136 TI - Interventional Radiology for Treating Soft Tissue Sarcomas. AB - The role of interventional radiology for soft tissue sarcomas is only occasionally addressed in the literature. However, different techniques such as embolization, selective chemotherapy, chemoembolization, and acrylic cement osteoplasty can be helpful with the primary tumor, recurrences, and metastases. This article discusses these techniques and their complications in treatment of soft tissue sarcomas. PMID- 11387137 TI - MR Imaging in the Follow-Up Evaluation of Aggressive Soft Tissue Tumors. AB - Although newer imaging techniques aid in the diagnosis of soft tissue sarcomas and current surgical procedures and adjunct therapy decrease the incidence of their recurrences when patients are referred to oncologic centers, the majority of these tumors are suboptimally evaluated in general practice with frequent recurrences. Close surveillance of these patients is necessary for early recognition and treatment of recurrent disease. This article describes an organized, stepwise magnetic resonance imaging evaluation of these patients after surgery and radiation therapy. The initial sequence should be T2-weighted. Low signal intensity or diffuse high signal intensity without mass on T2-weighted sequence excludes recurrence in 99% of patients. A high signal intensity mass, however, should be studied with bolus injection of contrast medium. Although standard contrast-enhanced MR imaging differentiates the non-enhancing hygroma or hematoma from the enhancing recurrence or pseudotumor, dynamic contrast-enhanced MR imaging is required to distinguish the rapidly enhancing recurrent tumor from the slowly enhancing pseudotumor of inflammation. PMID- 11387138 TI - An Algorithm for Soft Tissue Tumors: Current Perspectives. AB - The diagnosis and treatment of soft tissue sarcomas has dramatically changed in the last 10 to 15 years with more precise preoperative diagnosis, resection, adjunct therapy, and long-term follow-up. With correct preoperative diagnosis and staging of a soft tissue sarcoma, the surgeon, in conjunction with the radiation therapist and the medical oncologist, can select the optimal therapeutic approach to minimize the development of recurrences and metastases. Recognition of a mass as benign or non-neoplastic prevents unnecessary surgery. This article presents an algorithm that demonstrates the important interactions and contributions of the various oncologic-related disciplines (surgery, radiology, pathology, medical oncology, and radiation therapy) to the care of patients with soft tissue tumors. PMID- 11387139 TI - Proximal Femoral Focal Deficiency(PFFD): More Than An Abnormal Hip. AB - As the name implies, proximal femoral focal deficiency (PFFD) is a failure in development of the proximal femur and acetabulum of varying degrees. This article reviews the classification schemes with illustrated examples. Clinical findings, associated anomalies, imaging, and treatment are discussed. Patients are classified by radiographs, often changing classes as the patientOs skeleton matures. Magnetic resonance imaging (MRI) can aid in earlier and more accurate classification. The classification scheme exists to predict future function and the role of surgical intervention. PMID- 11387140 TI - Fibular Hemimelia: More Than Just An Absent Bone. AB - Fibular hemimelia is a congenital deficiency or absence of the fibula. There is a spectrum of disease from mild fibular hypoplasia to fibular aplasia. The ipsilateral tibia may be hypoplastic, bowed or normal. Fibular hemimelia can be an isolated deformity of the lower leg but frequently it is associated with proximal focal femoral deficiency, deficiencies of the lateral aspect of the foot, or is part of a malformation syndrome. In this article, we review the embryology of the extremities, discuss proposed etiologies for fibular hemimelia, highlight associated abnormalities, and present the radiographic and imaging findings. Surgical treatment options and long-term outcomes are discussed. PMID- 11387141 TI - Tarsal Coalition in Pediatric Patients. AB - A fibrous, cartilaginous, or bony (or a combination thereof) connection between two tarsal bones, or tarsal coalition usually involves the calcaneonavicular or talocalcaneal articulations. Foot pain, beginning when an initially fibrous coalition becomes partially or totally osseous and coinciding with trauma or increased stress such as athletic activity, is the usual presenting symptom. Although often identified on radiographs of the foot (including specialized projections), tarsal coalitions are increasingly imaged with computed tomography of the feet. Plain tomography, single contrast arthrography, and bone scintigraphy are no longer used in the evaluation of tarsal coalition. Magnetic resonance imaging has some advantages over computed tomography but its exact role remains undetermined. Surgical resection of symptomatic tarsal coalitions is used when conservative methods fail to alleviate symptoms. PMID- 11387142 TI - Common Pediatric Sports Injuries. AB - This article reviews the common acute and overuse injuries encountered in the pediatric athlete. Acute injuries are usually physeal or avulsion fractures relating to a single traumatic event. Overuse injuries are the result of repetitive stress and include the common traction apophysitis, osteochondritis dissecans, and stress fractures. Sports-related injuries most frequently involve the lower extremity with injury patterns and frequencies relative to the athlete's age, size, and type of sport. Indeed, an alternative title for this review might be Othe adolescent athlete as the changing biomechanics and psychosocial stresses of adolescence are inherent risk factors for sports-related injuries. An estimated seven million adolescents currently play high school sports with an increasing number becoming interested in extreme sports. It is hoped that this review will assist your future encounters with the injured pediatric athlete or Oweekend warrior. PMID- 11387143 TI - Magnetic Resonance Imaging in Children with Scoliosis. AB - For many years, orthopaedic surgeons and radiologists have used plain-film radiography for diagnosis, treatment decision-making, and postsurgical evaluation of scoliosis. Recently, the direct visualization of the spinal canal and spinal cord became possible with the development of magnetic resonance imaging (MRI), which over a short period of time has become an indicated procedure in children with juvenile-onset idiopathic scoliosis and congenital scoliosis. Children with idiopathic scoliosis associated with neurologic findings on physical examination, with pain or rapid progression of the spine curvature are also being evaluated with MRI, especially if surgery for stabilization of the spine is planned. The use of MRI as a screening modality for all children with scoliosis is still controversial and further studies are needed to evaluate if the examination adds or changes the treatment of these patients. PMID- 11387144 TI - Scoliosis in Children: Surgical Management and Postoperative Radiographic Appearances. AB - A component of the evaluation of scoliosis in children is radiography of the spine, initially in anteroposterior and lateral projections. In selected circumstances, additional imaging of the neural axis with magnetic resonance imaging is indicated to identify or exclude an associated spinal cord or neural canal abnormality. The initial management of scoliosis is almost always conservative, employing bracing or casting. When conservative methods fail to control curvature progression, spinal surgery is indicated. A variety of instrumentation devices have been developed in order to reduce and stabilize spinal curvatures. This article will dicuss and illustrate the more frequently used instrumentation devices as well as other selected aspects of spinal surgery. PMID- 11387145 TI - Nuclear Medicine in Pediatric Musculoskeletal Imaging. AB - The pediatric skeleton is a dynamic structure. Bone scintigraphy depends on bone metabolism and therefore is particularly applicable to changes occurring in the physes (growth-plates). A knowledge of the physiologic changes in the growing skeleton as well as the disease processes affecting the different pediatric age brackets are necessary to accurately solve clinical diagnostic problems with bone scintigraphy. Bone scintigraphy is a readily available, low-radiation imaging procedure. In this article, the common pediatric skeletal aberrations (appendicular growth alterations, hip pain, back pain, some aspects of trauma, inflammatory and infectious processes, and some common benign neoplastic occurrences) that can be addressed by nuclear imaging are discussed. PMID- 11387146 TI - Gamma camera positron emission tomography. PMID- 11387147 TI - Bias in plain film reading performance studies. AB - Radiographers and other healthcare professionals are becoming increasingly involved in radiological reporting, for instance plain radiographs, mammography and ultrasound. Systematic reviews of research evidence can help to assimilate a knowledge base by ordering and evaluating the available evidence on the reporting accuracy of different professional groups. This article reviews the biases that can undermine the results of plain film reading performance studies. These biases are subdivided into three categories. The first category refers to the selection of subjects, including both films and professionals, and covers the validity of generalizing results beyond the study population. The other two categories are concerned with study design and the interpretation both of films and of reports and the effect on study validity. An understanding of these biases is essential when designing such studies and when interpreting the results of existing studies. PMID- 11387148 TI - Geographical distribution of breast cancers on the mammogram: an interval cancer database. AB - Auditing interval cancers is an important part of a breast screening radiologist's continuing education. We set out to determine whether the position of interval cancers on the mammogram differs from those detected at screening. The 773 interval cancers so far identified, and the first 200 screen detected cancers, have been entered onto a Microsoft Access 97 database developed to record pathological and radiological features, including the position of the cancer on a stylized diagram using a "point and click" system. Reports were generated showing positions of all interval cancers by classification and reader. The distribution of true interval cancers is statistically different from screen detected cancers on both views. The distribution of the false negative and screen detected cancers only differs on the oblique view. False negative and true interval cancers are of the same distribution on both craniocaudal and oblique views. However, these differences do not appear to be practically useful when applied to individual readers. We have developed a database that allows systematic recording of pathological and radiological information regarding breast cancers. Additionally, it can record the geographic position of the cancer with minimal memory requirements. Statistical differences in the distribution of false negative and screen detected cancers have been demonstrated and the stylized diagrams reinforce the importance of the conventional review areas. Although this has not identified any "blind spots" in our own readers, it nevertheless provides film readers with a tool to audit their own work. PMID- 11387149 TI - Development of a protocol for coronal reconstruction of the maxillofacial region from axial helical CT data. AB - Using a fresh frozen cadaver head, a series of axial helical CT scans were obtained using varying imaging parameters both before and after traumatizing the head. The appearance of reformatted coronal images was optimized for the lowest radiation dose. A protocol for imaging the maxillofacial region was developed that produced diagnostic coronal reconstructed images from the axial helical CT data. PMID- 11387150 TI - Identification of vertebral arteries on CT of the chest. AB - Knowledge of the superior mediastinal course of the vertebral arteries is important for radiologists who evaluate chest CT, particularly in the setting of trauma, when planning a percutaneous interventional procedure or for pre operative planning. Our aim was to determine how often the vertebral arteries could be identified on chest CT studies. Contrast enhanced chest CT studies from 100 consecutive patients were reviewed, with specific attention to the vertebral arteries in the superior mediastinal and thoracic outlet regions. The left vertebral artery was identified in 85 patients and the right vertebral artery in 76 patients. Non-visualization of a vertebral artery was usually owing to proximal venous occlusion with extensive collateral vessels in the expected location of the vertebral arteries, local lymphadenopathy, poor contrast bolus technique or local beam hardening artefact. Radiologists need to alert surgeons planning resection of mass lesions in this region to the location of the vertebral arteries. It is also important to note that a vertebral artery was not identified on chest CT in 24% of patients. PMID- 11387151 TI - Vesicoureteric reflux and renal scarring in Chinese children. AB - Vesicoureteric reflux (VUR) and renal scarring are commonly found in children with urinary tract infection (UTI). The prevalence of VUR and scarring may vary between racial groups. There are no published data on the prevalence of VUR and scarring in Chinese children with UTI. A retrospective, single-institution study was made of Hong Kong Chinese children aged less than 5 years with a documented UTI investigated by both micturating cystourethrography and dimercaptosuccinic acid scintigraphy. VUR was identified in 39% of 93 Chinese children with UTI. Renal scarring was present in 28% of boys, which is comparable with published data on Western children. Scarring appears to be less common in Chinese girls with UTI (11%) than in Western girls (30-38% from published data), and its severity is poorly related to VUR grade. There is a significant dependency relationship between grade of VUR and degree of scarring in Chinese boys (p < 0.05). In conclusion, renal scarring appears to be relatively uncommon in Chinese girls. The correlation between grade of VUR and degree of scarring in Chinese boys suggests a relationship, but provides no evidence about the direction of causation. PMID- 11387152 TI - A parametric method for determining mammographic X-ray tube output and half value layer. AB - In the National Health Service Breast Screening Programme, regular assessment of the mean glandular doses received by a group of women is recognized as an important part of a quality assurance programme. The use of different tube voltages, to improve the beam penetration for thick or dense breasts, and of X ray units with programmable exposure modes, requires a large number of measurements to ensure that all the values of tube output and half value layer required for the dose calculations are available. In this work, a computer model is used to produce data that allow the calculation of tube output and half value layer for the range of clinically encountered conditions from measurements routinely obtained during quality assurance tests. The data are given as a series of equations and parameters, enabling the calculations to be easily incorporated into a dose calculation spreadsheet. The results of an experimental verification of the model are presented, showing good agreement between the measured and predicted values of half value layer and tube output for a range of combinations of target, filter and tube voltage. PMID- 11387153 TI - Development of a phantom for morphometric X-ray absorptiometry. AB - Morphometric X-ray absorptiometry (MXA) has recently been developed to assess vertebral deformity status using dual energy X-ray absorptiometry (DXA) machines. In contrast to bone densitometry, a vertebral morphometry phantom is not supplied by any machine manufacturer. The aim of this study was to develop a suitable phantom to quantify the accuracy and precision of the vertebral measurement software on three DXA scanners in vitro and to perform a weekly quality control (QC) scan over a 30-month period to evaluate any drift or changes in measurement accuracy over time. The phantom was constructed from Perspex and aluminium to simulate soft tissue and bone, respectively. 13 aluminium rectangles (each 30 mm wide, 25 mm high and 3 mm thick, with edges ("endplates") 6 mm thick) were set into one side of a solid Perspex block to represent the vertebral bodies from the fourth thoracic (T4) to the fourth lumbar (L4). The phantom was scanned on both the Hologic QDR2000plus and the QDR-4500A as well as the Lunar Expert-XL. Three consecutive lateral MXA scans were acquired on the Hologic machines using each of the scan modes available. On the QDR-2000plus, the lateral scan modes available are fast, array and high definition, which are all dual energy modes. These three scan modes are also available on the QDR-4500A, with the addition of a single energy scan mode. Four lateral scans were acquired on the Expert-XL machine using the single scan mode available. Each MXA scan was analysed twice by a trained operator using the standard software supplied by each manufacturer. A QC scan was performed approximately weekly over a 30-month period on only the QDR-4500A machine, and total phantom height was measured from the inferior edge of L4 to the superior edge of T4. Accuracy of "vertebral" height measurement varied between the three DXA machines and between the scan modes available. All underestimated "true" vertebral height by between 0.4% and 8.6%, with the scan modes using finer collimation producing the most accurate results. Repeat analysis precision of vertebral height measurement was best on the QDR-4500A, followed by the Expert-XL, and was poorest on the QDR-2000plus. The QC scans acquired on the QDR-4500A suggested that it was a highly stable machine, little affected by even major repairs. It must be remembered that these in vitro phantom results may not be representative of the true in vivo situation. The MXA phantom appears to be a useful tool for documenting the stability of the mechanical instruments and for checking the long-term consistency of operator precision. PMID- 11387154 TI - The diagnostic X-ray protection characteristics of Panelcrete, Aquapanel, Betopan and Gypsoplak Superboard. AB - Panelcrete, Aquapanel and Betopan are cement-based building materials with uses similar to those of gypsum wallboard, whose properties as a diagnostic X-ray shielding material have been extensively studied. The X-ray attenuation characteristics of these cement-based boards as well as those of a gypsum wallboard, Gypsoplak Superboard, are investigated for broad beam geometry conditions and for tube potentials of 50 kVp, 70 kVp, 100 kVp, 125 kVp and 140 kVp. Comparisons between these materials as well as with published data for gypsum wallboard are made. An example of their use as secondary barriers is given. Furthermore, it is confirmed that when building materials are considered for diagnostic X-ray shielding, calculations based on data for similar materials and corrected for density differences can be used only as an approximation. PMID- 11387155 TI - The effect of beam tube potential variation on gonad dose to patients during chest radiography investigated using high sensitivity LiF:Mg,Cu,P thermoluminescent dosemeters. AB - Optimization of X-ray beam tube potential (kVp) in radiological examinations can minimize patient dose. This research aims to investigate the effect of tube potential variation on gonad doses to patients during posteroanterior (PA) chest radiography examinations. This study was carried out using a Toshiba general purpose X-ray unit and a Rando phantom. Dose measuring equipment included an ion chamber system, a dose-area product (DAP) meter and a thermoluminescent dosemeter (TLD) reader system with high sensitivity TLD pellets of LiF:Mg,Cu,P for low level gonad dose measurement. PA chest exposures of the phantom to produce a constant exit dose were made using a standard low tube potential (range 60-100 kVp) non-grid technique and a high tube potential (range 95-150 kVp) grid technique. Entrance surface doses (ESDs) and DAPs were also included in the measurements. Effective doses (EDs) were computed from ESD and DAP measurements using NRPB-SR262 and Xdose software. Results show that with the low tube potential technique both ovary dose and testes dose increase with increasing tube potential; statistically significant correlations of r = 0.994 (p = 0.0006) and r = 0.998 (p = 0.001), respectively, were found. For both organs, doses increase at a rate of approximately 2% per kVp. With the high tube potential technique there is insignificant correlation between gonad doses and tube potential. When comparing patient doses from typical exposures made at 70 kVp (low tube potential non-grid technique) with doses from exposures made at 120 kVp (high tube potential grid technique), the high tube potential technique delivers significantly higher values for ESD, and ovary, testes and effective doses by factors of 1.7, 5.2, 5.5 and 2.7, respectively. PMID- 11387156 TI - Radiotherapy combined with simultaneous chemotherapy with mitomycin-C and 5 fluorouracil for inoperable head and neck cancer. AB - The feasibility and effectiveness of a combined chemoradiotherapy treatment modality for locally advanced head and neck cancer was tested in a phase II trial. From March 1995 to June 1998, 35 patients with advanced squamous cell carcinoma of the head and neck were treated with a continuous intravenous infusion of 5-fluorouracil (600 mg m-2 24 h-1 for Days 1 to 5 (120 h)) and mitomycin-C (10 mg m-2 intravenously) on Day 5 during the first week of radiotherapy and on Day 36. 31 patients had stage IV disease; 4 patients had stage III; and 1 patient had stage II. Patient ages ranged from 42-69 years (median 56.7 years). The tumours involved were as follows: oral cavity (n = 11); oropharynx (n = 14); hypopharynx/larynx (n = 10). Radiotherapy was delivered to a total dose of 70 Gy with conventional fractionation (2 Gy per fraction, five times a week). Chemotherapy was well tolerated and all patients received the intended dose. Mild nausea occurred in five patients. After a mean follow-up of 21 months (range 10-44 months), 8 (23%) patients remain alive. A complete response was seen in 28 (80%) patients. When a recurrence appeared, it was within the first year after treatment. 1- and 2-year overall survival rates were 46% and 23%, respectively. Grade 3 mucositis occurred in 17% of patients. Grade 1-2 thrombopaenia occurred in 3 (9%) patients, grade > 2 leukopaenia in 4 (11%) patients, and grade > or = 2 anaemia in 3 (9%) patients. We observed a treatment interruption of maximum 1 week for six patients owing to mucositis. Febrile neutropaenia or aplasia were not observed. The concomitant use of 5-fluorouracil, mitomycin-C and radiotherapy in locally advanced head and neck carcinoma is well tolerated in this group of patients. This protocol showed good locoregional response with a very low toxicity profile. PMID- 11387157 TI - False aneurysm of the pancreaticoduodenal artery complicating therapeutic endoscopic retrograde cholangiopancreatography. AB - A 76-year-old woman underwent two endoscopic retrograde cholangiopancreatography (ERCP) procedures for palliation of a carcinoma of the pancreas. At the first procedure a pre-cut sphincterotomy was performed because deep cannulation of the biliary tree was impossible. An endoscopic plastic biliary stent was inserted at the second ERCP. The patient developed abdominal pain and a post-procedure CT demonstrated a pseudoaneurysm. This was not present on the pre-procedure CT and was thought to arise from the pancreaticoduodenal artery as a complication of the pre-cut sphincterotomy. Visceral angiography confirmed the origin of the aneurysm from a branch of the inferior pancreaticoduodenal artery. The aneurysm was successfully embolised. To our knowledge, this is the first time that this complication has been reported. PMID- 11387158 TI - Breast uptake of iodine-131 mimicking lung metastases in a thyroid cancer patient with a pituitary tumour. AB - We report a case of a 44-year-old female thyroid cancer patient with a pituitary tumour. 131I whole body scanning showed a change of chest uptake from a unilateral crescent uptake to a bilateral full breast uptake pattern. Hyperprolactinaemia and a pituitary tumour were diagnosed as a result of observing the 131I breast uptake. PMID- 11387159 TI - CT appearances of HIV-related lipodystrophy syndrome. AB - Highly active antiretroviral therapy in HIV-1 infected patients is associated with a lipodystrophy syndrome, characterized by wasting of peripheral fat, central adiposity, hyperlipidaemia and insulin resistance. The CT findings are presented and the differential diagnosis is discussed. PMID- 11387160 TI - Magnetic resonance evaluation of the pericardium. AB - Magnetic resonance (MR) is well suited to imaging the pericardium. High resolution images synchronized with the cardiac cycle can be obtained in any plane. The wide field of view allows additional anatomical and functional information to be obtained from adjacent structures such as the aorta, pleura, lungs and mediastinum. MR is particularly useful in cases of pericardial constriction without an associated effusion, in patients with complex or loculated pericardial effusions and in pericardial tumours. In this article we illustrate the characteristic MR features of a variety of pericardial pathologies. PMID- 11387161 TI - Case of the month. Pyrexia of unknown origin in an elderly lady. PMID- 11387162 TI - Managing spontaneous first trimester miscarriage. PMID- 11387163 TI - Caesarean section for fetal distress. PMID- 11387164 TI - Better standards for better reporting of RCTs. PMID- 11387165 TI - Do probiotics prevent childhood illnesses? PMID- 11387166 TI - BMJ bans "accidents". PMID- 11387167 TI - New global health fund. PMID- 11387168 TI - Talks from BMJ editors. PMID- 11387170 TI - EC to encourage closer European cooperation on cancer research. PMID- 11387169 TI - GMC chief retires as council votes for revalidation. PMID- 11387171 TI - Israelis hold Palestinian hospital chief. PMID- 11387175 TI - Meeker the second time around. PMID- 11387176 TI - Effect of long term consumption of probiotic milk on infections in children attending day care centres: double blind, randomised trial. AB - OBJECTIVE: To examine whether long term consumption of a probiotic milk could reduce gastrointestinal and respiratory infections in children in day care centres. DESIGN: Randomised, double blind, placebo controlled study over seven months. SETTING: 18 day care centres in Helsinki, Finland. PARTICIPANTS: 571 healthy children aged 1-6 years: 282 (mean (SD) age 4.6 (1.5) years) in the intervention group and 289 (mean (SD) age 4.4 (1.5) years) in the control group. INTERVENTION: Milk with or without Lactobacillus GG. Average daily consumption of milk in both groups was 260 ml. MAIN OUTCOME MEASURES: Number of days with respiratory and gastrointestinal symptoms, absences from day care because of illness, respiratory tract infections diagnosed by a doctor, and course of antibiotics. RESULTS: Children in the Lactobacillus group had fewer days of absence from day care because of illness (4.9 (95% confidence interval 4.4 to 5.5) v 5.8 (5.3 to 6.4) days, 16% difference, P=0.03; age adjusted 5.1 (4.6 to 5.6) v 5.7 (5.2 to 6.3) days, 11% difference, P=0.09). There was also a relative reduction of 17% in the number of children suffering from respiratory infections with complications and lower respiratory tract infections (unadjusted absolute % reduction -8.6 (-17.2 to -0.1), P=0.05; age adjusted odds ratio 0.75 (0.52 to 1.09), P=0.13) and a 19% relative reduction in antibiotic treatments for respiratory infection (unadjusted absolute % reduction -9.6 (-18.2 to -1.0), P=0.03; adjusted odds ratio 0.72 (0.50 to 1.03), P=0.08) in the Lactobacillus group. CONCLUSIONS: Lactobacillus GG may reduce respiratory infections and their severity among children in day care. The effects of the probiotic Lactobacillus GG were modest but consistently in the same direction. PMID- 11387177 TI - Interval between decision and delivery by caesarean section-are current standards achievable? Observational case series. AB - OBJECTIVES: To audit interval from decision to delivery in urgent caesarean section to determine whether the current standard of 30 minutes is achievable routinely; to determine whether delay leads to an excess of admissions to special care. DESIGN: Three audit cycles over four years followed by a continuous audit over 32 months. SETTING: Large district general hospital delivering 5500 women each year. PARTICIPANTS: All women delivered by urgent caesarean section for abnormal fetal heart rate patterns, cord prolapse, failed instrumental delivery, or suspected placental abruption. MAIN OUTCOME MEASURES: Proportion of women delivered within 30 and 40 minutes of decision. Admission rates to special care by length of interval between decision and delivery. RESULTS: In the continuous audit 478 of 721 (66.3%) women were delivered in 30 minutes and 637 (88.3%) within 40 minutes; 29 (4.0%) were undelivered at 50 minutes. If the woman was taken to theatre in 10 minutes, 409 of 500 (81.8%) were delivered in 30 minutes and 495 (97%) in 40 minutes. There was no significant difference in the proportion of babies born at 36 weeks or later who were admitted to special care, when analysed by interval from decision to delivery. 36/449 (8%) babies with an interval from decision to delivery of less than 30 minutes were admitted to special care and 3/23 (13%) with an interval of more than 50 minutes were admitted. CONCLUSIONS: The current recommendations for the interval between decision and delivery are not being achieved in routine practice. Failure to meet the recommendations does not seem to increase neonatal morbidity. PMID- 11387178 TI - Prospective 12 month study of 30 minute decision to delivery intervals for "emergency" caesarean section. PMID- 11387179 TI - Comparison of breastfeeding rates in Scotland in 1990-1 and 1997-8. PMID- 11387180 TI - Analysis of trends in premature mortality by Labour voting in the 1997 general election. PMID- 11387182 TI - Cluster randomised controlled trial to compare three methods of promoting secondary prevention of coronary heart disease in primary care. AB - OBJECTIVE: To assess the effectiveness of three different methods of promoting secondary prevention of coronary heart disease in primary care. DESIGN: Pragmatic, unblinded, cluster randomised controlled trial. SETTING: Warwickshire. SUBJECTS: 21 general practices received intervention; outcome measured in 1906 patients aged 55-75 years with established coronary heart disease. INTERVENTIONS: Audit of notes with summary feedback to primary health care team (audit group); assistance with setting up a disease register and systematic recall of patients to general practitioner (GP recall group); assistance with setting up a disease register and systematic recall of patients to a nurse led clinic (nurse recall group). MAIN OUTCOME MEASURES: At 18 months' follow up: adequate assessment (defined) of 3 risk factors (blood pressure, cholesterol, and smoking status); prescribing of hypotensive agents, lipid lowering drugs, and antiplatelet drugs; blood pressure, serum cholesterol level, and plasma cotinine levels. RESULTS: Adequate assessment of all 3 risk factors was much more common in the nurse and GP recall groups (85%, 76%) than the audit group (52%). The advantage in the nurse recall compared with the audit group was 33% (95% confidence interval 19% to 46%); in the GP recall group compared with the audit group 23% (10% to 36%), and in the nurse recall group compared with the GP recall group 9% (-3% to 22%). However, these differences in assessment were not reflected in clinical outcomes. Mean blood pressure (148/80, 147/81, 148/81 mm Hg), total cholesterol (5.4, 5.5, 5.5 mmol/l), and cotinine levels (% probable smokers 17%, 16%, 19%) varied little between the nurse recall, GP recall, and audit groups respectively, as did prescribing of hypotensive and lipid lowering agents. Prescribing of antiplatelet drugs was higher in the nurse recall group (85%) than the GP recall or audit groups (80%, 74%). After adjustment for baseline levels, the advantage in the nurse recall group compared with the audit group was 10% (3% to 17%), in the nurse recall group compared with the GP recall group 8% (1% to 15%) and in the GP recall group compared with the audit group 2% (-6% to 10%). CONCLUSIONS: Setting up a register and recall system improved patient assessment at 18 months' follow up but was not consistently better than audit alone in improving treatment or risk factor levels. Understanding the reasons for this is the key next step in improving the quality of care of patients with coronary heart disease. PMID- 11387184 TI - Management of spontaneous miscarriage in the first trimester: an example of putting informed shared decision making into practice. PMID- 11387186 TI - Lesson of the week: Playing the odds in clinical decision making: lessons from berry aneurysms undetected by magnetic resonance angiography. PMID- 11387187 TI - ABC of AIDS: Treatment of infections. PMID- 11387188 TI - Why should women have lower reference limits for haemoglobin and ferritin concentrations than men? PMID- 11387189 TI - Are quality of life measures patient centred? PMID- 11387196 TI - A structural pathway for activation of the kinesin motor ATPase. AB - Molecular motors move along actin or microtubules by rapidly hydrolyzing ATP and undergoing changes in filament-binding affinity with steps of the nucleotide hydrolysis cycle. It is generally accepted that motor binding to its filament greatly increases the rate of ATP hydrolysis, but the structural changes in the motor associated with ATPase activation are not known. To identify the conformational changes underlying motor movement on its filament, we solved the crystal structures of three kinesin mutants that decouple nucleotide and microtubule binding by the motor, and block microtubule-activated, but not basal, ATPase activity. Conformational changes in the structures include a disordered loop and helices in the switch I region and a visible switch II loop, which is disordered in wild-type structures. Switch I moved closer to the bound nucleotide in two mutant structures, perturbing water-mediated interactions with the Mg2+. This could weaken Mg2+ binding and accelerate ADP release to activate the motor ATPASE: The structural changes we observe define a signaling pathway within the motor for ATPase activation that is likely to be essential for motor movement on microtubules. PMID- 11387197 TI - Increased epidermal tumors and increased skin wound healing in transgenic mice overexpressing the catalytic subunit of telomerase, mTERT, in basal keratinocytes. AB - Telomerase transgenics are an important tool to assess the role of telomerase in cancer, as well as to evaluate the potential use of telomerase for gene therapy of age-associated diseases. Here, we have targeted the expression of the catalytic component of mouse telomerase, mTERT, to basal keratinocytes using the bovine keratin 5 promoter. These telomerase-transgenic mice are viable and show histologically normal stratified epithelia with high levels of telomerase activity and normal telomere length. Interestingly, the epidermis of these mice is highly responsive to the mitogenic effects of phorbol esters, and it is more susceptible than that of wild-type littermates to the development skin tumors upon chemical carcinogenesis. The epidermis of telomerase-transgenic mice also shows an increased wound-healing rate compared with wild-type littermates. These results suggest that, contrary to the general assumption, telomerase actively promotes proliferation in cells that have sufficiently long telomeres and unravel potential risks of gene therapy for age-associated diseases based on telomerase upregulation. PMID- 11387198 TI - Thrombospondin-2 plays a protective role in multistep carcinogenesis: a novel host anti-tumor defense mechanism. AB - The angiogenic switch during tumorigenesis is thought to be induced by a change in the balance of pro- angiogenic and anti-angiogenic factors. To elucidate the biological role of the endogenous angiogenesis inhibitor thrombospondin-2 (TSP-2) during multistep carcinogenesis, we subjected TSP-2-deficient and wild-type mice to a chemical skin carcinogenesis regimen. Surprisingly, TSP-2 expression was strongly upregulated in the mesenchymal stroma of wild-type mice throughout the consecutive stages of tumorigenesis whereas the angiogenesis factor, vascular endothelial growth factor, was induced predominantly in tumor cells. TSP-2 deficiency dramatically enhanced susceptibility to skin carcinogenesis and resulted in accelerated and increased tumor formation. The angiogenic switch occurred in early stages of pre-malignant tumor formation, and tumor angiogenesis was significantly enhanced in TSP-2-deficient mice. While TSP-2 deficiency did not affect tumor differentiation or proliferation, tumor cell apoptosis was significantly reduced. These results reveal upregulation of an endogenous angiogenesis inhibitor during multi step tumorigenesis and identify enhanced stromal TSP-2 expression as a novel host anti-tumor defense mechanism. PMID- 11387199 TI - CD40 and LMP-1 both signal from lipid rafts but LMP-1 assembles a distinct, more efficient signaling complex. AB - CD40, a member of the TNFR-1 receptor family, shares several features with LMP-1, an oncoprotein encoded by Epstein-Barr virus. CD40 and LMP-1 activate transcription by binding to TRAFs, JAK3 and/or TRADD. CD40's association with CD40L activates signaling. However, LMP-1 signals independently of a ligand but dependently on self-association. We demonstrate that activated CD40 and LMP-1 co localize in lipid rafts and recruit TRAF3 there, findings consistent with signals of CD40 and LMP-1 being initiated from lipid rafts. To elucidate their signaling, we compared requirements for their aggregation and subcellular localization. Targeting CD40's monomeric C-terminal signaling domain to lipid rafts activates signaling, as does rendering it trimeric. Addition of both modifications supports signaling more efficiently. Parallel experiments with LMP-1 indicate that targeting the monomeric C-terminal signaling domain of LMP-1 to lipid rafts activates signaling, but trimerizing it does not. Fusing LMP-1's N-terminus and membrane-spanning domains to CD40's C-terminus supports signaling more efficiently than CD40 plus ligand or CD40's trimerized and/or localized derivatives. An activity of LMP-1's N-terminus and membrane-spanning domains other than trimerization must contribute to its efficient signaling. PMID- 11387200 TI - C26-CoA-dependent ceramide synthesis of Saccharomyces cerevisiae is operated by Lag1p and Lac1p. AB - Lag1p and Lac1p are two highly homologous membrane proteins of the endoplasmic reticulum (ER). When both genes are deleted, cells cannot transport glycosylphosphatidylinositol (GPI)-anchored proteins from the ER to the Golgi at a normal rate. Here we show that microsomes or detergent extracts from lag1lac1 double mutants lack an activity transferring C26 fatty acids from C26-coenzyme A onto dihydrosphingosine or phytosphingosine. As a consequence, in intact cells, the normal ceramides and inositolphosphorylceramides are drastically reduced. lag1lac1 cells compensate for the lack of normal sphingolipids by making increased amounts of C26 fatty acids, which become incorporated into glycerophospholipids. They also contain 20- to 25-fold more free long chain bases than wild type and accumulate very large amounts of abnormally polar ceramides. They make small amounts of abnormal mild base-resistant inositolphospholipids. The lipid remodelling of GPI-anchored proteins is severely compromised in lag1lac1 double mutants since only few and mostly abnormal ceramides are incorporated into the GPI anchors. The participation of Lag1p and Lac1p in ceramide synthesis may explain their role in determining longevity. PMID- 11387202 TI - Energized mitochondria increase the dynamic range over which inositol 1,4,5 trisphosphate activates store-operated calcium influx. AB - In eukaryotic cells, activation of cell surface receptors that couple to the phosphoinositide pathway evokes a biphasic increase in intracellular free Ca2+ concentration: an initial transient phase reflecting Ca2+ release from intracellular stores, followed by a plateau phase due to Ca2+ influx. A major component of this Ca2+ influx is store-dependent and often can be measured directly as the Ca2+ release-activated Ca2+ current (I(CRAC)). Under physiological conditions of weak intracellular Ca2+ buffering, respiring mitochondria play a central role in store-operated Ca2+ influx. They determine whether macroscopic I(CRAC) activates or not, to what extent and for how long. Here we describe an additional role for energized mitochondria: they reduce the amount of inositol 1,4,5-trisphosphate (InsP3) that is required to activate I(CRAC). By increasing the sensitivity of store-operated influx to InsP3, respiring mitochondria will determine whether modest levels of stimulation are capable of evoking Ca2+ entry or not. Mitochondrial Ca2+ buffering therefore increases the dynamic range of concentrations over which the InsP3 is able to function as the physiological messenger that triggers the activation of store operated Ca2+ influx. PMID- 11387201 TI - NAADP induces Ca2+ oscillations via a two-pool mechanism by priming IP3- and cADPR-sensitive Ca2+ stores. AB - In sea urchin eggs, Ca2+ mobilization by nicotinic acid adenine dinucleotide phosphate (NAADP) potently self-inactivates but paradoxically induces long-term Ca2+ oscillations. We investigated whether NAADP-induced Ca2+ oscillations arise from the recruitment of other Ca2+ release pathways. NAADP, inositol trisphosphate (IP3) and cyclic ADP-ribose (cADPR) all mobilized Ca2+ from internal stores but only NAADP consistently induced Ca2+ oscillations. NAADP induced Ca2+ oscillations were partially inhibited by heparin or 8-amino-cADPR alone, but eliminated by the presence of both, indicating a requirement for both IP3- and cADPR-dependent Ca2+ release. Thapsigargin completely blocked IP3 and cADPR responses as well as NAADP-induced Ca2+ oscillations, but only reduced the NAADP-mediated Ca2+ transient. Following NAADP-mediated release from this Ca2+ pool, the amount of Ca2+ in the Ca2+-induced Ca2+ release stores was increased. These results support a mechanism in which Ca2+ oscillations are initiated by Ca2+ release from NAADP-sensitive Ca2+ stores (pool 1) and perpetuated through cycles of Ca2+ uptake into and release from Ca2+-induced Ca2+ release stores (pool 2). These results provide the first direct evidence in support of a two pool model for Ca2+ oscillations. PMID- 11387203 TI - Plasticity and adaptation of Ca2+ signaling and Ca2+-dependent exocytosis in SERCA2(+/-) mice. AB - Darier's disease (DD) is a high penetrance, autosomal dominant mutation in the ATP2A2 gene, which encodes the SERCA2 Ca2+ pump. Here we have used a mouse model of DD, a SERCA2(+/-) mouse, to define the adaptation of Ca2+ signaling and Ca2+ dependent exocytosis to a deletion of one copy of the SERCA2 gene. The [Ca2+]i transient evoked by maximal agonist stimulation was shorter in cells from SERCA2(+/-) mice, due to an up-regulation of specific plasma membrane Ca2+ pump isoforms. The change in cellular Ca2+ handling caused approximately 50% reduction in [Ca2+]i oscillation frequency. Nonetheless, agonist-stimulated exocytosis was identical in cells from wild-type and SERCA2(+/-) mice. This was due to adaptation in the levels of the Ca2+ sensors for exocytosis synaptotagmins I and III in cells from SERCA2(+/-) mice. Accordingly, exocytosis was approximately 10 fold more sensitive to Ca2+ in cells from SERCA2(+/-) mice. These findings reveal a remarkable plasticity and adaptability of Ca2+ signaling and Ca2+-dependent cellular functions in vivo, and can explain the normal function of most physiological systems in DD patients. PMID- 11387204 TI - The Ca2+ concentration of the endoplasmic reticulum is a key determinant of ceramide-induced apoptosis: significance for the molecular mechanism of Bcl-2 action. AB - The mechanism of action of the anti-apoptotic oncogene Bcl-2 is still largely obscure. We have recently shown that the overexpression of Bcl-2 in HeLa cells reduces the Ca2+ concentration in the endoplasmic reticulum ([Ca2+]er) by increasing the passive Ca2+ leak from the organelle. To investigate whether this Ca2+ depletion is part of the mechanism of action of Bcl-2, we mimicked the Bcl-2 effect on [Ca2+]er by different pharmacological and molecular approaches. All conditions that lowered [Ca2+]er protected HeLa cells from ceramide, a Bcl-2 sensitive apoptotic stimulus, while treatments that increased [Ca2+]er had the opposite effect. Surprisingly, ceramide itself caused the release of Ca2+ from the endoplasmic reticulum and thus [Ca2+] increased both in the cytosol and in the mitochondrial matrix, paralleled by marked alterations in mitochondria morphology. The reduction of [Ca2+]er levels, as well as the buffering of cytoplasmic [Ca2+] changes, prevented mitochondrial damage and protected cells from apoptosis. It is therefore concluded that the Bcl-2-dependent reduction of [Ca2+]er is an important component of the anti-apoptotic program controlled by this oncogene. PMID- 11387205 TI - The death substrate Gas2 binds m-calpain and increases susceptibility to p53 dependent apoptosis. AB - Gas2 is a caspase-3 substrate that plays a role in regulating microfilament and cell shape changes during apoptosis. Here we provide evidence that overexpression of Gas2 efficiently increases cell susceptibility to apoptosis following UV irradiation, etoposide and methyl methanesulfonate treatments, and that these effects are dependent on increased p53 stability and transcription activity. To investigate possible pathways linking Gas2 to p53, a yeast two-hybrid screen swas performed, indicating m-calpain as a strong Gas2- interacting protein. Moreover, we demonstrate that Gas2 physically interacts with m-calpain in vivo and that recombinant Gas2 inhibits calpain-dependent processing of p53. Importantly, the Gas2 dominant-negative form (Gas2171-314) that binds calpain but is unable to inhibit its activity abrogates Gas2's ability to stabilize p53, to enhance p53 transcriptional activity and to induce p53-dependent apoptosis. Finally, we show that Gas2 is able to regulate the levels of p53 independently of Mdm2 status, suggesting that, like calpastatin, it may enhance p53 stability by inhibiting calpain activity. PMID- 11387206 TI - Netrin-1 acts as a survival factor via its receptors UNC5H and DCC. AB - The membrane receptors DCC and UNC5H have been shown to be crucial for axon guidance and neuronal migration by acting as receptors for netrin-1. DCC has also been proposed as a dependence receptor inducing apoptosis in cells that are beyond netrin-1 availability. Here we show that the netrin-1 receptors UNC5H (UNC5H1, UNC5H2, UNC5H3) also act as dependence receptors. UNC5H receptors induce apoptosis, but this effect is blocked in the presence of netrin-1. Moreover, we demonstrate that UNC5H receptors are cleaved in vitro by caspase in their intracellular domains. This cleavage may lead to the exposure of a fragment encompassing a death domain required for cell death induction in vivo. Finally, we present evidence that during development of the nervous system, the presence of netrin-1 is crucial to maintain survival of UNC5H- and DCC-expressing neurons, especially in the ventricular zone of the brainstem. Altogether, these results argue for a role of netrin-1 during the development of the nervous system, not only as a guidance cue but as a survival factor via its receptors DCC and UNC5H. PMID- 11387207 TI - Ezrin is a downstream effector of trafficking PKC-integrin complexes involved in the control of cell motility. AB - Protein kinase C (PKC) alpha has been implicated in beta1 integrin-mediated cell migration. Stable expression of PKCalpha is shown here to enhance wound closure. This PKC-driven migratory response directly correlates with increased C-terminal threonine phosphorylation of ezrin/moesin/radixin (ERM) at the wound edge. Both the wound migratory response and ERM phosphorylation are dependent upon the catalytic function of PKC and are susceptible to inhibition by phosphatidylinositol 3-kinase blockade. Upon phorbol 12,13-dibutyrate stimulation, green fluorescent protein-PKCalpha and beta1 integrins co-sediment with ERM proteins in low-density sucrose gradient fractions that are enriched in transferrin receptors. Using fluorescence lifetime imaging microscopy, PKCalpha is shown to form a molecular complex with ezrin, and the PKC-co-precipitated endogenous ERM is hyperphosphorylated at the C-terminal threonine residue, i.e. activated. Electron microscopy showed an enrichment of both proteins in plasma membrane protrusions. Finally, overexpression of the C-terminal threonine phosphorylation site mutant of ezrin has a dominant inhibitory effect on PKCalpha induced cell migration. We provide the first evidence that PKCalpha or a PKCalpha associated serine/threonine kinase can phosphorylate the ERM C-terminal threonine residue within a kinase-ezrin molecular complex in vivo. PMID- 11387208 TI - SKP1-SnRK protein kinase interactions mediate proteasomal binding of a plant SCF ubiquitin ligase. AB - Arabidopsis Snf1-related protein kinases (SnRKs) are implicated in pleiotropic regulation of metabolic, hormonal and stress responses through their interaction with the kinase inhibitor PRL1 WD-protein. Here we show that SKP1/ASK1, a conserved SCF (Skp1-cullin-F-box) ubiquitin ligase subunit, which suppresses the skp1-4 mitotic defect in yeast, interacts with the PRL1-binding C-terminal domains of SnRKs. The same SnRK domains recruit an SKP1/ASK1-binding proteasomal protein, alpha4/PAD1, which enhances the formation of a trimeric SnRK complex with SKP1/ASK1 in vitro. By contrast, PRL1 reduces the interaction of SKP1/ASK1 with SnRKs. SKP1/ASK1 is co-immunoprecipitated with a cullin SCF subunit (AtCUL1) and an SnRK kinase, but not with PRL1 from Arabidopsis cell extracts. SKP1/ASK1, cullin and proteasomal alpha-subunits show nuclear co-localization in differentiated Arabidopsis cells, and are observed in association with mitotic spindles and phragmoplasts during cell division. Detection of SnRK in purified 26S proteasomes and co-purification of epitope- tagged SKP1/ASK1 with SnRK, cullin and proteasomal alpha-subunits indicate that the observed protein interactions between SnRK, SKP1/ASK1 and alpha4/PAD1 are involved in proteasomal binding of an SCF ubiquitin ligase in Arabidopsis. PMID- 11387209 TI - Activated MEK5 induces serial assembly of sarcomeres and eccentric cardiac hypertrophy. AB - Mitogen-activated protein kinase (MAPK) pathways couple intrinsic and extrinsic signals to hypertrophic growth of cardiomyocytes. The MAPK kinase MEK5 activates the MAPK ERK5. To investigate the potential involvement of MEK5-ERK5 in cardiac hypertrophy, we expressed constitutively active and dominant-negative forms of MEK5 in cardiomyocytes in vitro. MEK5 induced a form of hypertrophy in which cardiomyocytes acquired an elongated morphology and sarcomeres were assembled in a serial manner. The cytokine leukemia inhibitory factor (LIF), which stimulates MEK5 activity, evoked a similar response. Moreover, a dominant-negative MEK5 mutant specifically blocked LIF-induced elongation of cardiomyocytes and reduced expression of fetal cardiac genes without blocking other aspects of LIF-induced hypertrophy. Consistent with the ability of MEK5 to induce serial assembly of sarcomeres in vitro, cardiac-specific expression of activated MEK5 in transgenic mice resulted in eccentric cardiac hypertrophy that progressed to dilated cardiomyopathy and sudden death. These findings reveal a specific role for MEK5 ERK5 in the induction of eccentric cardiac hypertrophy and in transduction of cytokine signals that regulate serial sarcomere assembly. PMID- 11387210 TI - A single autophosphorylation site on KDR/Flk-1 is essential for VEGF-A-dependent activation of PLC-gamma and DNA synthesis in vascular endothelial cells. AB - KDR/Flk-1 tyrosine kinase, one of the two vascular endothelial growth factor (VEGF) receptors, induces mitogenesis and differentiation of vascular endothelial cells. To understand the mechanisms underlying the VEGF-A-induced growth signaling pathway, we constructed a series of human KDR mutants and examined their biological properties. An in vitro kinase assay and subsequent tryptic peptide mapping revealed that Y1175 and Y1214 are the two major VEGF-A-dependent autophosphorylation sites. Using an antibody highly specific to the phosphoY1175 region, we demonstrated that Y1175 is phosphorylated rapidly in vivo in primary endothelial cells. When the mutated KDRs were introduced into the endothelial cell lines by adenoviral vectors, only the Y1175F KDR, Tyr1175 to phenylalanine mutant, lost the ability to tyrosine phosphorylate phospholipase C-gamma and, significantly, reduced MAP kinase phosphorylation and DNA synthesis in response to VEGF-A. Furthermore, primary endothelial cells microinjected with anti phosphoY1175 antibody clearly decreased DNA synthesis compared with control cells. These findings strongly suggest that autophosphorylation of Y1175 on KDR is crucial for endothelial cell proliferation, and that this region is a new target for anti-angiogenic reagents. PMID- 11387211 TI - Arabidopsis thaliana Rop GTPases are localized to tips of root hairs and control polar growth. AB - Plants contain a novel unique subfamily of Rho GTPases, vital components of cellular signalling networks. Here we report a general role for some members of this family in polarized plant growth processes. We show that Arabidopsis AtRop4 and AtRop6 encode functional GTPases with similar intrinsic GTP hydrolysis rates. We localized AtRop proteins in root meristem cells to the cross-wall and cell plate membranes. Polar localization of AtRops in trichoblasts specifies the growth sites for emerging root hairs. These sites were visible before budding and elongation of the Arabidopsis root hair when AtRops accumulated at their tips. Expression of constitutively active AtRop4 and AtRop6 mutant proteins in root hairs of transgenic Arabidopsis plants abolished polarized growth and delocalized the tip-focused Ca2+ gradient. Polar localization of AtRops was inhibited by brefeldin A, but not by other drugs such as latrunculin B, cytochalasin D or caffeine. Our results demonstrate a general function of AtRop GTPases in tip growth and in polar diffuse growth. PMID- 11387212 TI - The adaptor molecule Disabled-2 links the transforming growth factor beta receptors to the Smad pathway. AB - Using a genetic complementation approach we have identified disabled-2 (Dab2), a structural homolog of the Dab1 adaptor molecule, as a critical link between the transforming growth factor beta (TGFbeta) receptors and the Smad family of proteins. Expression of wild-type Dab2 in a TGFbeta-signaling mutant restores TGFbeta-mediated Smad2 phosphorylation, Smad translocation to the nucleus and Smad-dependent transcriptional responses. TGFbeta stimulation triggers a transient increase in association of Dab2 with Smad2 and Smad3, which is mediated by a direct interaction between the N-terminal phosphotyrosine binding domain of Dab2 and the MH2 domain of Smad2. Dab2 associates with both the type I and type II TGFbeta receptors in vivo, suggesting that Dab2 is part of a multiprotein signaling complex. Together, these data indicate that Dab2 is an essential component of the TGFbeta signaling pathway, aiding in transmission of TGFbeta signaling from the TGFbeta receptors to the Smad family of transcriptional activators. PMID- 11387213 TI - Hormone-induced nucleosome positioning in the MMTV promoter is reversible. AB - The mouse mammary tumor virus (MMTV) promoter is induced by glucocorticoid hormone via the glucocorticoid receptor (GR). The hormone-triggered effects on MMTV transcription and chromatin structure were studied in Xenopus oocytes. We previously showed that the nucleosomes organizing the MMTV promoter became translationally positioned upon hormone induction. A single GR-binding site was necessary and sufficient for the chromatin events to occur, while transcription and basal promoter elements were dispensable. Here we show that addition of the hormone antagonists RU486 or RU43044 to the previously hormone-induced MMTV promoter results in cessation of transcription and loss of chromatin remodeling and nucleosome positioning. In vivo footprinting demonstrated agonist- and RU486 induced GR binding to its DNA response element (GRE), while the other antagonist, RU43044, did not promote GR-GRE interaction. These results demonstrate that induction and maintenance of nucleosome positioning is an active process that requires constant 'pressure' of agonist-GR-recruited chromatin-modifying factor(s) rather than GR-DNA binding itself. PMID- 11387214 TI - Binding of Ikaros to the lambda5 promoter silences transcription through a mechanism that does not require heterochromatin formation. AB - The Ikaros family of proteins are DNA binding factors required for correct development of B and T lymphocytes. Cytogenetic studies have shown that these proteins form complexes with pericentromeric heterochromatin in B cells, and the colocalization of transcriptionally silent genes with these complexes suggests that Ikaros could silence transcription by recruiting genes to heterochromatin. Here we show that a site in the lambda5 promoter that binds Ikaros and Aiolos is required for silencing of lambda5 expression in activated mature B cells. Analysis of methylation and nuclease accessibility indicates that the silenced lambda5 gene is not heterochromatinized in B cells, despite being associated with pericentromeric heterochromatin clusters. We also found that a promoter mutation, which affects Ikaros-mediated silencing of lambda5 expression, is not rescued in a transgenic line that has the gene integrated into pericentromeric heterochromatin. Our results indicate that the Ikaros proteins initiate silencing of lambda5 expression through a direct effect on the promoter with localization to pericentromeric heterochromatin likely to affect the action of Ikaros on regulatory sequences rather than causing heterochromatinization of the gene. PMID- 11387215 TI - The RNA polymerase III transcription initiation factor TFIIIB participates in two steps of promoter opening. AB - Evidence for post-recruitment functions of yeast transcription factor (TF)IIIB in initiation of transcription was first provided by the properties of TFIIIB-RNA polymerase III-promoter complexes assembled with deletion mutants of its Brf and B" subunits that are transcriptionally inactive because they fail to open the promoter. The experiments presented here show that these defects can be repaired by unpairing short (3 or 5 bp) DNA segments spanning the transcription bubble of the open promoter complex. Analysis of this suppression phenomenon indicates that TFIIIB participates in two steps of promoter opening by RNA polymerase III that are comparable to the successive steps of promoter opening by bacterial RNA polymerase holoenzyme. B" deletions between amino acids 355 and 421 interfere with the initiating step of DNA strand separation at the upstream end of the transcription bubble. Removing an N-terminal domain of Brf interferes with downstream propagation of the transcription bubble to and beyond the transcriptional start site. PMID- 11387216 TI - Heme mediates derepression of Maf recognition element through direct binding to transcription repressor Bach1. AB - Heme controls expression of genes involved in the synthesis of globins and heme. The mammalian transcription factor Bach1 functions as a repressor of the Maf recognition element (MARE) by forming antagonizing hetero-oligomers with the small Maf family proteins. We show here that heme binds specifically to Bach1 and regulates its DNA-binding activity. Deletion studies demonstrated that a heme binding region of Bach1 is confined within its C-terminal region that possesses four dipeptide cysteine-proline (CP) motifs. Mutations in all of the CP motifs of Bach1 abolished its interaction with heme. The DNA-binding activity of Bach1 as a MafK hetero-oligomer was markedly inhibited by heme in gel mobility shift assays. The repressor activity of Bach1 was lost upon addition of hemin in transfected cells. These results suggest that increased levels of heme inactivate the repressor Bach1, resulting in induction of a host of genes with MARES: PMID- 11387217 TI - Inability to enter S phase and defective RNA polymerase II CTD phosphorylation in mice lacking Mat1. AB - The trimeric Cdk7-cyclin H-Mat1 complex comprises the kinase subunit of basal transcription factor TFIIH and has been shown to function as a cyclin-dependent kinase (Cdk)-activating kinase. Herein we report that disruption of the murine Mat1 gene leads to peri-implantation lethality coincident with depletion of maternal Mat1 protein. In culture, Mat1(-/-) blastocysts gave rise to viable post mitotic trophoblast giant cells while mitotic lineages failed to proliferate and survive. In contrast to wild-type trophoblast giant cells, Mat1(-/-) cells exhibited a rapid arrest in endoreduplication, which was characterized by an inability to enter S phase. Additionally, Mat1(-/-) cells exhibited defects in phosphorylation of the C-terminal domain (CTD) of RNA polymerase II on both Ser5 and Ser2 of the heptapeptide repeat. Despite this, Mat1(-/-) cells demonstrated apparent transcriptional and translational integrity. These data indicate an essential role for Mat1 in progression through the endocycle and suggest that while Mat1 modulates CTD phosphorylation, it does not appear to be essential for RNA polymerase II-mediated transcription. PMID- 11387218 TI - A role for DNA polymerase alpha in epigenetic control of transcriptional silencing in fission yeast. AB - In the fission yeast Schizosaccharomyces pombe, transcriptional silencing at the mating-type region, centromeres and telomeres is epigenetically controlled, and results from the assembly of higher order chromatin structures. Chromatin proteins associated with these silenced loci are believed to serve as molecular bookmarks that help promote inheritance of the silenced state during cell division. Specifically, a chromodomain protein Swi6 is believed to be an important determinant of the epigenetic imprint. Here, we show that a mutation in DNA polymerase alpha (pol(alpha)) affects Swi6 localization at the mating-type region and causes a 45-fold increase in spontaneous transition from the silenced epigenetic state to the expressed state. We also demonstrate that pol(alpha) mutant cells are defective in Swi6 localization at centromeres and telomeres. Genetic analysis suggests that Polalpha and Swi6 are part of the same silencing pathway. Interestingly, we found that Swi6 directly binds to Pol(alpha) in vitro. Moreover, silencing-defective mutant Pol(alpha) displays reduced binding to Swi6 protein. This work indicates involvement of a DNA replication protein, Pol(alpha), in heterochromatin assembly and inheritance of epigenetic chromatin structures. PMID- 11387219 TI - Human acrocentric chromosomes with transcriptionally silent nucleolar organizer regions associate with nucleoli. AB - Human ribosomal gene repeats are distributed among five nucleolar organizer regions (NORs) on the p arms of acrocentric chromosomes. On exit from mitosis, nucleoli form around individual active NORs. As cells progress through the cycle, these mini-nucleoli fuse to form large nucleoli incorporating multiple NORs. It is generally assumed that nucleolar incorporation of individual NORs is dependent on ribosomal gene transcription. To test this assumption, we determined the nuclear location of individual human acrocentric chromosomes, and their associated NORs, in mouse> human cell hybrids. Human ribosomal genes are transcriptionally silent in this context. Combined immunofluorescence and in situ hybridization (immuno-FISH) on three-dimensional preserved nuclei showed that human acrocentric chromosomes associate with hybrid cell nucleoli. Analysis of purified nucleoli demonstrated that human and mouse NORs are equally likely to be within a hybrid cell nucleolus. This is supported further by the observation that murine upstream binding factor can associate with human NORs. Incorporation of silent NORs into mature nucleoli raises interesting issues concerning the maintenance of the activity status of individual NORs. PMID- 11387220 TI - Requirement of Hsp90 for centrosomal function reflects its regulation of Polo kinase stability. AB - We have previously shown that the molecular chaperone heat shock protein 90 (Hsp90) is required to ensure proper centrosome function in Drosophila and vertebrate cells. This observation led to the hypothesis that this chaperone could be required for the stability of one or more centrosomal proteins. We have found that one of these is Polo, a protein kinase known to regulate several aspects of cell division including centrosome maturation and function. Inhibition of Hsp90 results in the inactivation of Polo kinase activity. It also leads to a loss in the ability of cytoplasmic extracts to complement the failure of salt stripped preparations of centrosomes to nucleate microtubules. This effect can be rescued upon addition of active recombinant POLO: We also show that Polo and Hsp90 are part of a complex and conclude that stabilization of Polo is one of the mechanisms by which Hsp90 contributes to the maintenance of functional centrosomes. PMID- 11387221 TI - Cell cycle transition under stress conditions controlled by vertebrate heat shock factors. AB - The roles of heat shock transcription factors (HSFs) under physiological conditions have recently become the focus of intense study. We generated avian cells lacking two heat-inducible HSFs, HSF1 and HSF3. In addition to complete loss of activation of heat shock genes under stress conditions, these cells exhibited a marked reduction in Hsp90alpha expression under normal growth conditions. Reduction in Hsp90alpha expression caused instability of a cyclin dependent kinase, Cdc2, and cell cycle progression was blocked mainly at the G2 phase, but also at G1 phase even at mild heat shock temperatures. Restoration of Hsp90alpha expression rescued the temperature sensitivity without induction of HSPS: We demonstrated for the first time a molecular target affected by heat shock in vivo that causes cell cycle arrest in vertebrates and a novel mechanism of stress resistance controlled by vertebrate HSFS: PMID- 11387222 TI - Silent repair accounts for cell cycle specificity in the signaling of oxidative DNA lesions. AB - Reactive oxygen species are the most important source of DNA lesions in aerobic organisms, but little is known about the activation of the DNA checkpoints in response to oxidative stress. We show that treatment of yeast cells with sublethal concentrations of hydrogen peroxide induces a Mec1-dependent phosphorylation of Rad53 and a Rad53-dependent cell cycle delay specifically during S phase. The lack of Rad53 phosphorylation after hydrogen peroxide treatment in the G1 and G2 phases is due to the silent repair of oxidative DNA lesions produced at these stages by the base excision repair (BER) pathway. Only the disruption of the BER pathway and the accumulation and/or treatment of DNA intermediates by alternative repair pathways reveal the existence of primary DNA lesions induced at all phases of the cell cycle by hydrogen peroxide. Our data illustrate both the concept of silent repair of DNA damage and the high sensitivity of S-phase cells to hydrogen peroxide. PMID- 11387223 TI - Engineered interphase chromosome loops guide intrachromosomal recombination. AB - How large-scale topologies regulate interphase chromosome function remains an important question in eukaryotic cell biology. Looped structures are thought to modulate transcription by pairing promoters with distant control elements and to orchestrate intrachromosomal recombination events by pairing appropriate recombination partners. To explore the effects of chromosomal topology on intrachromosomal recombination, distinct loop geometries were engineered into chromosome III of the budding yeast Saccharomyces cerevisiae. These topologies were created by employing pairs of lac operator clusters to serve as pairing sites and a modified lac repressor to perform the role of a protein cross-bridge. The influence of these engineered loops on the selection of donor loci during mating-type switching was evaluated using novel genetic and molecular methods. These experiments demonstrate that engineered interphase chromosome loops are biologically active-capable of influencing the course of intrachromosomal recombination. They also provide insight into the mechanism of mating-type switching by revealing a causal relationship between defined chromosomal topologies and the choice of donor locus. PMID- 11387224 TI - Altered nucleotide misinsertion fidelity associated with poliota-dependent replication at the end of a DNA template. AB - A hallmark of human DNA polymerase iota (poliota) is the asymmetric fidelity of replication at template A and T when the enzyme extends primers annealed to a single-stranded template. Here, we report on the efficiency and accuracy of poliota-dependent replication at a nick, a gap, the very end of a template and from a mispaired primer. Poliota cannot initiate synthesis on a nicked DNA substrate, but fills short gaps efficiently. Surprisingly, poliota's ability to blunt-end a 1 bp recessed terminus is dependent upon the template nucleotide encountered and is highly erroneous. At template G, both C and T are inserted with roughly equal efficiency, whilst at template C, C and A are misinserted 8- and 3-fold more often than the correct base, G. Using substrates containing mispaired primer termini, we show that poliota can extend all 12 mispairs, but with differing efficiencies. Poliota can also extend a tandem mispair, especially when it is located within a short gap. The enzymatic properties of poliota appear consistent with that of a somatic hypermutase and suggest that poliota may be one of the low-fidelity DNA polymerases hypothesized to participate in the hypermutation of immunoglobulin variable genes in vivo. PMID- 11387225 TI - Tipping the balance between replicative and simple transposition. AB - The bacterial insertion sequence IS903 has the unusual ability to transpose both replicatively and non-replicatively. The majority of products are simple insertions, while co-integrates, the product of replicative transposition, occur at a low frequency (<0.1% of simple insertions). In order to define the critical steps that determine the outcome of IS903 transposition, we have isolated mutants that specifically increase the rate of replicative transposition. Here we show that the nucleotide immediately flanking the transposon influences both overall transposition frequency and co-integrate formation. In particular, when the 3' flanking nucleotide is A, co-integrates are increased 500-fold compared with a 3' C. In addition, we have isolated five transposase mutants that increase replicative transposition. These residues are close to the catalytic residues and are thus likely to be part of the active site. These are the first transposase mutations described that affect the product of transposition. Our results are consistent with the hypothesis that a delay in cleavage of the 5'-flanking DNA will increase the effective half-life of the 3'-nicked transposon intermediate and consequently enhance co-integrate formation. PMID- 11387226 TI - Determinants for hairpin formation in Tn10 transposition. AB - Tn10 transposition involves the formation of a hairpin intermediate at the transposon termini. Here we show that hairpin formation exhibits more stringent DNA sequence requirements at the terminal two base pairs than either transpososome assembly or first strand nicking. We also observe a significant DNA distortion at the terminal base pairs upon transpososome assembly by chemical nuclease footprinting. Interestingly, mutations at these positions do not necessarily inhibit the formation of the distortion. However, it remains a possibility that the inhibitory effect of these mutations is due to a defect in protein-DNA interactions subsequent to this deformation. Terminal base pair mutations also inhibited strand transfer, providing evidence that transposase interactions with the terminal residues on both 'transferred' and 'non transferred' strands are important for hairpin formation. We also demonstrate that mutation of a highly conserved tyrosine residue that is a component of the YREK motif, Y285, results in a phenotype comparable to that of the terminal base pair mutations. In contrast, a mutation at another conserved position, W265, is shown to relax the specificity of the hairpin formation reaction. PMID- 11387227 TI - RNomics: an experimental approach that identifies 201 candidates for novel, small, non-messenger RNAs in mouse. AB - In mouse brain cDNA libraries generated from small RNA molecules we have identified a total of 201 different expressed RNA sequences potentially encoding novel small non-messenger RNA species (snmRNAs). Based on sequence and structural motifs, 113 of these RNAs can be assigned to the C/D box or H/ACA box subclass of small nucleolar RNAs (snoRNAs), known as guide RNAs for rRNA. While 30 RNAs represent mouse homologues of previously identified human C/D or H/ACA snoRNAs, 83 correspond to entirely novel snoRNAS: Among these, for the first time, we identified four C/D box snoRNAs and four H/ACA box snoRNAs predicted to direct modifications within U2, U4 or U6 small nuclear RNAs (snRNAs). Furthermore, 25 snoRNAs from either class lacked antisense elements for rRNAs or snRNAS: Therefore, additional snoRNA targets have to be considered. Surprisingly, six C/D box snoRNAs and one H/ACA box snoRNA were expressed exclusively in brain. Of the 88 RNAs not belonging to either snoRNA subclass, at least 26 are probably derived from truncated heterogeneous nuclear RNAs (hnRNAs) or mRNAS: Short interspersed repetitive elements (SINEs) are located on five RNA sequences and may represent rare examples of transcribed SINES: The remaining RNA species could not as yet be assigned either to any snmRNA class or to a part of a larger hnRNA/mRNA. It is likely that at least some of the latter will represent novel, unclassified snmRNAS: PMID- 11387228 TI - A subcomplex of three eIF3 subunits binds eIF1 and eIF5 and stimulates ribosome binding of mRNA and tRNA(i)Met. AB - Yeast translation initiation factor 3 contains five core subunits (known as TIF32, PRT1, NIP1, TIF34 and TIF35) and a less tightly associated component known as HCR1. We found that a stable subcomplex of His8-PRT1, NIP1 and TIF32 (PN2 subcomplex) could be affinity purified from a strain overexpressing these eIF3 subunits. eIF5, eIF1 and HCR1 co-purified with this subcomplex, but not with distinct His8-PRT1- TIF34-TIF35 (P45) or His8-PRT1-TIF32 (P2) sub complexes. His8 PRT1 and NIP1 did not form a stable binary subcomplex. These results provide in vivo evidence that TIF32 bridges PRT1 and NIP1, and that eIFs 1 and 5 bind to NIP1, in native eIF3. Heat-treated prt1-1 extracts are defective for Met tRNA(i)Met binding to 40S subunits, and we also observed defective 40S binding of mRNA, eIFs 1 and 5 and eIF3 itself in these extracts. We could rescue 40S binding of Met- tRNA(i)Met and mRNA, and translation of luciferase mRNA, in a prt1-1 extract almost as well with purified PN2 subcomplex as with five-subunit eIF3, whereas the P45 subcomplex was nearly inactive. Thus, several key functions of eIF3 can be carried out by the PRT1-TIF32-NIP1 subcomplex. PMID- 11387229 TI - Transfer RNA(Ala) recognizes transfer-messenger RNA with specificity; a functional complex prior to entering the ribosome? AB - tmRNA (SsrA or 10Sa RNA) functions as both a transfer RNA and a messenger RNA, rescues stalled ribosomes and clears the cell of incomplete polypeptides. We report that native Escherichia coli tmRNA interacts specifically with native or synthetic E.coli tRNA alanine (tRNA(Ala)) in vitro, alanine being the first codon of the tmRNA internal open reading frame. Aminoacylatable RNA microhelices also bind tmRNA. Complex formation was monitored by gel retardation assays combined with structural probes. Nucleotides from the acceptor stem of tRNA(Ala) are essential for complex formation with tmRNA. tRNA(Ala) isoacceptors recognize tmRNA with different affinities, with an important contribution from tRNA(Ala) post-transcriptional modifications. The most abundant tRNA(Ala) isoacceptor in vivo binds tmRNA with the highest affinity. A complex between tRNA(Ala) and tmRNA might involve up to 140 tmRNA molecules out of 500 present per E.coli cell. Our data suggest that tmRNA interacts with the tRNA that decodes the resume codon prior to entering the ribosome. Biological implications of promoting specific complexes between tmRNA and aminoacylatable RNAs are discussed, with emphasis on primitive versions of the translation apparatus. PMID- 11387230 TI - Specific interaction between the ribosome recycling factor and the elongation factor G from Mycobacterium tuberculosis mediates peptidyl-tRNA release and ribosome recycling in Escherichia coli. AB - Once the translating ribosomes reach a termination codon, the nascent polypeptide chain is released in a factor-dependent manner. However, the P-site-bound deacylated tRNA and the ribosomes themselves remain bound to the mRNA (post termination complex). The ribosome recycling factor (RRF) plays a vital role in dissociating this complex. Here we show that the Mycobacterium tuberculosis RRF (MtuRRF) fails to rescue Escherichia coli LJ14, a strain temperature-sensitive for RRF (frr(ts)). More interestingly, co-expression of M.tuberculosis elongation factor G (MtuEFG) with MtuRRF rescues the frr(ts) strain of E.coli. The simultaneous expression of MtuEFG is also needed to cause an enhanced release of peptidyl-tRNAs in E.coli by MtuRRF. These observations provide the first genetic evidence for a functional interaction between RRF and EFG. Both the in vivo and in vitro analyses suggest that RRF does not distinguish between the translating and terminating ribosomes for their dissociation from mRNA. In addition, complementation of E.coli PEM100 (fusA(ts)) with MtuEFG suggests that the mechanism of RRF function is independent of the translocation activity of EFG. PMID- 11387231 TI - Current concepts in the pathogenesis of alcoholic liver injury. AB - Alcoholic liver disease (ALD) develops as a consequence of priming and sensitizing mechanisms rendered by cross-interactions of primary mechanistic factors and secondary risk factors. This concept, albeit not novel, is becoming widely accepted by the field, and more research is directed toward identifying and characterizing the interfaces of the cross-interactions to help understand individual predisposition to the disease. Another pivotal development is the beginning of cell type-specific research to elucidate specific contributions not only of hepatocytes, but also of hepatic macrophages, liver-associated lymphocytes, sinusoidal endothelial cells, and hepatic stellate cells to sensitizing and priming mechanisms. In particular, the critical role of hepatic macrophages has been highlighted and the priming mechanisms concerning this paracrine effect have been proposed. Glutathione depletion in hepatocyte mitochondria is considered the most important sensitizing mechanism. One of the contributing factors is decreased methionine metabolism. Remaining key questions include how altered methionine metabolism contribute to the pathogenesis of ALD; how cross-talk among nonparenchymal liver cells or between nonparenchymal cells and hepatocytes leads to ALD; how dysfunctional mitochondria determine the type of cell death in ALD; and what secondary factors are critical for the development of advanced ALD such as alcoholic hepatitis and cirrhosis. PMID- 11387232 TI - Growth hormone reduces plasma cholesterol in LDL receptor-deficient mice. AB - Growth hormone (GH) has pleiotropic effects on cholesterol and lipoprotein metabolism. Pituitary GH is important for the normal regulation of hepatic LDL receptors (LDLR), for the enzymatic activity of bile acid regulatory cholesterol 7alpha-hydroxylase (C7alphaOH), and for the maintenance of resistance to dietary cholesterol. The present study aimed to determine whether GH has beneficial effects on plasma lipids and hepatic cholesterol metabolism in mice devoid of LDLR. Compared with wild-type controls, LDLR-deficient mice had approximately 250% elevated plasma total cholesterol and approximately 50% increased hepatic cholesterol levels; hepatic HMG CoA reductase activity was reduced by 70%, whereas C7alphaOH activity was increased by 40%. In LDLR mice, GH infusion reduced plasma cholesterol and triglycerides up to 40%, whereas HMG CoA reductase and C7alphaOH activities were stimulated by approximately 50% and 110% respectively. GH also stimulated HMG CoA reductase and C7alphaOH activities in control mice, whereas hepatic LDLR and plasma lipoproteins were unchanged. The effects of cholestyramine and atorvastatin on C7alphaOH in LDLR-deficient mice were potentiated by GH, and this was associated with a further reduction in plasma cholesterol. GH treatment reduces plasma cholesterol and triglycerides and stimulates C7alphaOH activity in mice devoid of LDLR, particularly in combination with resin or statin treatment. The potential of GH therapy in patients with homozygous familial hypercholesterolemia should be evaluated. PMID- 11387233 TI - Insulin and glucocorticoids differentially regulate leptin transcription and secretion in brown adipocytes. AB - Leptin, the ob gene product, is produced by adipose tissue and is submitted to a complex hormonal and metabolic regulation. Leptin plays a critical role in the balance of body weight. Here we report on secretion and hormonal regulation of leptin by brown adipocytes. Using the recently established T37i cell line, we show that leptin expression and secretion occurred as a function of cell differentiation. In differentiated T37i cells, insulin induced leptin release ( approximately 0.25 ng/10(6) cells/h) in a concentration-dependent manner (EC50=0.1 nM), and this was totally suppressed by beta3-adrenergic ligand, thiazolidinedione, cycloheximide, or actinomycin D. Insulin induced a strong, rapid (within 2 h) but transient fivefold increase in leptin mRNA levels. This transcriptional control of ob gene expression by insulin involved both phosphatidylinositol 3-kinase- and MAP kinase-dependent pathways. Glucocorticoids inhibited both insulin-stimulated leptin secretion and ob gene expression without affecting leptin mRNA stability (t(1/2)=3h05). Altogether, our results demonstrate that brown adipocytes express and secrete leptin, whose hormonal regulation clearly differs from that described in white adipose tissue. These findings point to tissue-specific molecular mechanisms and suggest that leptin might exert direct effects on energy homeostasis through an autocrine mechanism. PMID- 11387234 TI - Developmental expression analysis of thyroid hormone receptor isoforms reveals new insights into their essential functions in cardiac and skeletal muscles. AB - Nuclear thyroid hormone (TH) receptors (TR) play a critical role in mediating the diverse actions of TH in development, differentiation, and metabolism of most tissues, but the role of TR isoforms in muscle development and function is unclear. Therefore, we have undertaken a comprehensive expression analysis of TRalpha 1, TRbeta 1, TRbeta 2 (TH binding), and TRalpha 2 (non-TH binding) in functionally distinct porcine muscles during prenatal and postnatal development. Use of a novel and highly sensitive RNase protection assay revealed striking muscle-specific developmental profiles of all four TR isoform mRNAs in cardiac, longissimus, soleus, rhomboideus, and diaphragm. Distribution of TR isoforms varied markedly between muscles; TRalpha expression was considerably greater than TRbeta and there were significant differences in the ratios TRalpha 1:TRalpha 2, and TRbeta 1:TRbeta 2. Together with immunohistochemistry of myosin heavy chain isoforms and data on myogenesis and maturation of the TH axis, these findings provide new evidence that highlights central roles for 1) TRalpha isoforms in fetal myogenesis, 2) the ratio TRalpha 1:TRalpha 2 in determining cardiac and skeletal muscle phenotype and function; 3) TRbeta in maintaining a basal level of cellular response to TH throughout development and a specific maturational function around birth. These findings suggest that events disrupting normal developmental profiles of TR isoforms may impair optimal function of cardiac and skeletal muscles. PMID- 11387235 TI - Activation of paracrine TGF-beta1 signaling upon stimulation and degranulation of rat serosal mast cells: a novel function for chymase. AB - As a source of transforming growth factor beta1 (TGF-beta1), mast cells have been implicated as potential effector cells in many pathological processes. However, the mechanisms by which mast cells express, secrete, and activate TGF-beta1 have remained vague. We show here by means of RT-PCR, immunoblotting, and immunocytochemistry that isolated rat peritoneal mast cells synthesize and store large latent TGF-beta1 in their chymase 1-containing secretory granules. Mast cell stimulation and degranulation results in rapid secretion of the latent TGF beta1, which is converted by chymase 1 into an active form recognized by the type II TGF-beta serine/threonine kinase receptor (TbetaRII). Thus, mast cells secrete active TGF-beta1 by a unique secretory mechanism in which latent TGF-beta1 and the activating enzyme chymase 1 are coreleased. The activation of latent TGF beta1 specifically by chymase was verified using recombinant human latent TGF beta1 and recombinant human chymase. In isolated TbetaRI- and TbetaRII-expressing peritoneal macrophages, the activated TGF-beta1 induces the expression of the plasminogen activator inhibitor 1 (PAI-1), whereas in the mast cells, the levels of TbetaRI, TbetaRII, and PAI-1 expression were below detection. Selective stimulation of mast cells in vivo in the rat peritoneal cavity leads to rapid overexpression of TGF-beta1 in peritoneal mast cells and of TbetaRs in peritoneal macrophages. These data strongly suggest that mast cells can act as potent paracrine effector cells both by secreting active TGF-beta1 and by enhancing its response in target cells. PMID- 11387236 TI - An angiogenic laminin site and its antagonist bind through the alpha(v)beta3 and alpha5beta1 integrins. AB - Angiogenesis is important for wound healing, tumor growth, and metastasis. Endothelial cells differentiate into capillary-like structures on a laminin-1 rich matrix (Matrigel). We previously identified 20 angiogenic sites on laminin-1 (alpha1beta1gamma1) by screening 559 overlapping synthetic peptides. C16, the most potent gamma1 chain peptide, blocked laminin-1-mediated adhesion and was the only gamma1 chain peptide to block attachment to both collagen I and fibronectin. This suggested that C16 was acting via a receptor common to these substrates. We demonstrated that C16 is angiogenic in vivo. Affinity chromatography identified the integrins alpha5beta1 and alpha(v)beta3 as surface receptors. Blocking antibodies confirmed the role of these receptors in C16 adhesion. C16 does not contain an RGD sequence and, as expected, an RGD-containing peptide did not block C16 adhesion nor did C16 act via MAP kinase phosphorylation. Furthermore, we identified a C16 scrambled sequence, C16S, which antagonizes the angiogenic activity of bFGF and of C16 by binding to the same receptors. Because the laminin gamma1 chain is ubiquitous in most tissues, C16 is likely an important functional site. Since the biological activity of C16 is blocked by a scrambled peptide, C16S may serve as an anti-angiogenic therapeutic agent. PMID- 11387237 TI - Hypothesis: inappropriate colonization of the premature intestine can cause neonatal necrotizing enterocolitis. AB - Neonatal necrotizing enterocolitis (NEC) is a major cause of morbidity in preterm infants. We hypothesize that the intestinal injury in this disease is a consequence of synergy among three of the major risk factors for NEC: prematurity, enteral feeding, and bacterial colonization. Together these factors result in an exaggerated inflammatory response, leading to ischemic bowel necrosis. Human milk may decrease the incidence of NEC by decreasing pathogenic bacterial colonization, promoting growth of nonpathogenic flora, promoting maturation of the intestinal barrier, and ameliorating the proinflammatory response. PMID- 11387238 TI - Lack of p53 accelerates hepatocarcinogenesis in transgenic mice constitutively overexpressing c-myc in the liver. PMID- 11387239 TI - Amyloid beta-peptide induces nitric oxide production in rat hippocampus: association with cholinergic dysfunction and amelioration by inducible nitric oxide synthase inhibitors. PMID- 11387240 TI - The complement system is an integrated part of the natural innate immune response in the brain. PMID- 11387241 TI - TNF-alpha regulates early differentiation of C2C12 myoblasts in an autocrine fashion. PMID- 11387242 TI - The cytoplasmic truncated receptor tyrosine kinase ALK homodimer immortalizes and cooperates with ras in cellular transformation. PMID- 11387243 TI - NADH: sensor of blood flow need in brain, muscle, and other tissues. PMID- 11387244 TI - Distinct heparan sulfate glycosaminoglycans are responsible for mediating fibroblast growth factor-2 biological activity through different fibroblast growth factor receptors. PMID- 11387245 TI - In vivo supplementation with coenzyme Q10 enhances the recovery of human lymphocytes from oxidative DNA damage. PMID- 11387246 TI - Induction and suppression of interferon-inducible protein 10 in reperfused myocardial infarcts may regulate angiogenesis. PMID- 11387247 TI - The synthesis of antimicrobial peptides in the skin of Rana esculenta is stimulated by microorganisms. PMID- 11387248 TI - Pharmacological dissection of vascular effects caused by activation of protease activated receptors 1 and 2 in anesthetized rats. PMID- 11387249 TI - Wnt regulation of limb mesenchymal chondrogenesis is accompanied by altered N cadherin-related functions. PMID- 11387250 TI - Functional mitochondria are required for amyloid beta-mediated neurotoxicity. PMID- 11387251 TI - Establishment of normal, terminally differentiating mouse erythroid progenitors: molecular characterization by cDNA arrays. PMID- 11387252 TI - The HPV16 E6 and E7 proteins and the radiation resistance of cervical carcinoma. PMID- 11387253 TI - ER60/ERp57 forms disulfide-bonded intermediates with MHC class I heavy chain. PMID- 11387254 TI - Cellular translocation of proteins by transportan. PMID- 11387255 TI - HMG-CoA reductase inhibition protects the diabetic myocardium from ischemia reperfusion injury. PMID- 11387256 TI - Involvement of alpha3 integrin/tetraspanin complexes in the angiogenic response induced by angiotensin II. PMID- 11387257 TI - Turnover of oxidatively damaged nuclear proteins in BV-2 microglial cells is linked to their activation state by poly-ADP-ribose polymerase. PMID- 11387258 TI - Effects of carboxyl-terminal fragment of Alzheimer's amyloid precursor protein and amyloid beta-peptide on the production of cytokines and nitric oxide in glial cells. PMID- 11387259 TI - Increased cAMP levels and protein kinase (PKA) type I activation in CD4+ T cells and B cells contribute to retrovirus-induced immunodeficiency of mice (MAIDS): a useful in vivo model for drug testing. PMID- 11387260 TI - A novel interleukin 13 (IL-13) antagonist that blocks the biological activity of human IL-13 in immune and nonimmune cells. PMID- 11387261 TI - Inactivation of glutathione peroxidase by NO leads to the accumulation of H2O2 and the induction of HB-EGF via c-Jun NH2-terminal kinase in rat aortic smooth muscle cells. PMID- 11387262 TI - Up-regulation of prosaposin by the retinoid HPR and its effect on ceramide production and integrin receptors. PMID- 11387263 TI - Heat-induced nuclear accumulation of hsc70s is regulated by phosphorylation and inhibited in confluent cells. PMID- 11387264 TI - Selenium deficiency increases the pathology of an influenza virus infection. PMID- 11387265 TI - Vasopressin receptor distribution in the liver controls calcium wave propagation and bile flow. PMID- 11387266 TI - The bradykinin/B1 receptor promotes angiogenesis by up-regulation of endogenous FGF-2 in endothelium via the nitric oxide synthase pathway. PMID- 11387267 TI - Post-transcriptional regulation of the AT1 receptor mRNA. Identification of the mRNA binding motif and functional characterization. PMID- 11387268 TI - Assessment of ovarian reserve - is there a role for ovarian biopsy? AB - The pool of primordial follicles in the ovary or 'ovarian reserve' is a major factor in the human fertility potential. The ageing ovary is characterized by reduction of the number of primordial follicles and this loss accelerates in the late 30's and precedes the menopause by 10-12 years. Woman's age alone or with a combination of biochemical markers, dynamic tests and ultrasound measurements fail to predict this loss accurately. In this manuscript, a novel approach of histopathological examination of ovarian biopsy for the evaluation of infertility, especially in unexplained infertility and in women in the later part of reproductive life, is discussed. PMID- 11387269 TI - Is there a physiological role for gonadotrophin oligosaccharide heterogeneity in humans? III. Luteinizing hormone heterogeneity: a medical physiologist's perspective. AB - Clinical evidence for a physiological impact of luteinizing hormone (LH) isoforms includes their unequal in-vitro bioactivity and altered in-vivo LH kinetics. For example, alkaline LH isospecies emerge in an oestrogen-rich milieu, and show greater bioactivity in vitro along with more rapid metabolic removal in vivo. More acidic LH isotypes are predominant in eugonadal men with end-stage renal failure and in postmenopausal women. The relevance of changes in charge distribution in puberty to sexual maturation is not clear. Molecular LH variants may be associated with decreased testis size and reduced linear growth in boys, menstrual irregularity and/or subfertility in women, and possibly protect against polycystic ovarian syndrome (PCOS). This article summarises the provisional physiological implications of LH isotypes based on current evidence. PMID- 11387270 TI - Focal vascular endothelial growth factor correlates with angiogenesis in human endometrium. Role of intravascular neutrophils. AB - Vascular endothelial growth factor (VEGF) is expressed in human endometrium, but the cellular source of VEGF for endometrial angiogenesis has not been determined. In the present study the relationship between focal VEGF associated with microvessels and endothelial cell proliferation was examined in three layers of human endometrium at various stages of the menstrual cycle (menstrual, proliferative and secretory). Immunohistochemical analysis of full thickness endometrium from 18 hysterectomy samples without endometrial pathology were examined. The percentage of proliferating vessels was higher in proliferative compared to secretory endometrium, but this was only statistically significant in the basalis layer. A significantly greater percentage of VEGF-expressing microvessels was observed in proliferative than secretory endometrium (P < 0.05). The most VEGF-expressing microvessels were observed in the subepithelial capillary plexus, followed by the functionalis and least were present in the basalis. There was a significant correlation between focal VEGF-expressing microvessels and proliferating vessels for the subepithelial capillary plexus (R(s) = 0.70, P = 0.008), the functionalis (R(s) = 0.70, P = 0.001) and the basalis (R(s) = 0.76, P < 0.001). Focal VEGF associated with microvessels was found in marginating and adherent neutrophils. These data suggest that neutrophils in intimate contact with endometrial endothelium may be a source of intravascular VEGF for vessels undergoing angiogenesis by elongation or intussusception, particularly during the proliferative phase of rapid endometrial growth. PMID- 11387271 TI - Follicular fluid as a favourable environment for endometrial and endometriotic cell growth in vitro. AB - Follicular fluid from women with endometriosis has been shown to induce a higher endometrial cell proliferation than that derived from women without the disease. To elucidate this issue further, the aims of the present study were to compare the ability of follicular fluid and peritoneal fluid to stimulate both endometrial and endometriotic cell proliferation and to verify whether the mitogenic effect was merely sex steroid-dependent. Endometrial and endometriotic cells were cultured in follicular fluid or peritoneal fluid diluted in serum-free media; the growth induced in these conditions was compared with that obtained by culturing these cells in medium supplemented with charcoal stripped calf serum and a correspondent content of 17-beta-oestradiol and progesterone. Follicular fluid was able to induce significantly higher cell proliferation than peritoneal fluid from controls, patients with endometriosis stage I-II and women with endometriosis stage III-IV (P < 0.05). Moreover, the growth in control media containing a corresponding amount of steroid hormones was significantly lower than that obtained with follicular or peritoneal fluids. This finding indicates that the stimulating effect is not simply related to the concentrations of 17 beta-oestradiol and progesterone present in these fluids. Finally, based on these results and on other previous observations, the hypothesis that follicular fluid may be involved in the development of endometriotic ovarian cysts is discussed. PMID- 11387272 TI - The data sources which may help strengthen the epidemiological evidence for the hormonal hypothesis of sex determination in man. AB - The hypothesis that parental hormone levels around the time of conception partially control offspring sex ratios-though here taken to be true in substance will need a great deal of work to specify with any accuracy. We do not know with any certainty which hormones are involved, nor how they are implicated. Answers to these two questions are only likely to emerge after prolonged experimental work. And it is fair to say that that work has not yet started. I assume that experimental workers will not embark on such a project until it is perfectly clear that there is a watertight case that mammalian parental hormone levels somehow influence offspring sex ratios. The present note indicates where further (human) evidence for that case will be found. In regard to human beings, much of the required information is held by clinics and registries not primarily concerned with reproductive biology. This point is illustrated here in regard to toxicology, teratology, radiation medicine, neurology, psychiatry, oncology, dermatology, rheumatology, occupational medicine and sports medicine as well as obstetrics and gynaecology. Tests (based on the hypothesis) are offered for intrauterine endocrine causes of malformations, and for pre- and post-natal endocrine causes of disease. PMID- 11387273 TI - The impact of obesity and insulin resistance on the outcome of IVF or ICSI in women with polycystic ovarian syndrome. AB - The impact of insulin resistance on the outcome of IVF or intracytoplasmic sperm injection (ICSI) in women with polycystic ovarian syndrome (PCOS) was examined. Insulin sensitivity was measured by the continuous infusion of glucose with model assessment (CIGMA) test. Insulin-resistant (n = 26) and non-insulin-resistant women (n = 30) with PCOS underwent a total of 100 cycles of long-term down regulation with buserelin acetate, stimulation with human recombinant FSH, and IVF or ICSI. Blood samples were taken throughout ovarian stimulation for hormone assays. Insulin-resistant and non-insulin-resistant women had similar concentrations of FSH, LH, testosterone and androstenedione throughout stimulation, but insulin-resistant women had hyperinsulinaemia and lower sex hormone binding globulin concentrations. Insulin-resistant women also had lower oestradiol concentrations during stimulation and required higher FSH doses, but these differences disappeared after controlling for the higher body weight in the group of insulin-resistant women. Groups had similar number of oocytes collected, similar implantation and pregnancy rates, and the incidence of ovarian hyperstimulation syndrome was also similar. Obesity, independent of hyperinsulinaemia, was related to a lower oocyte count and increased FSH requirement. It is concluded that in PCOS women receiving long-term down regulation and stimulation with recombinant FSH, insulin resistance is neither related to hormone levels nor to IVF outcome. Obesity, independent of insulin resistance, is associated with relative gonadotrophin resistance. PMID- 11387274 TI - The effect of gonadotrophins with differing LH/FSH ratios on the secretion of the various species of inhibin in women receiving IVF. AB - We have measured secretory patterns of inhibin A, B, total alpha inhibin, pro alphaC inhibin and oestradiol in women following pituitary suppression who were randomised into two groups to receive either urinary gonadotrophin (25:75 IU/ampoule of luteinizing hormone (LH) and follicle stimulating hormone (FSH; Normegon; n = 11) or recombinant (r)FSH (75 IU/ampoule of FSH alone, n = 16). The women were of similar age (approximately 33 years) and length of infertility (approximately 4 years) and had a normal endocrine evaluation. Plasma FSH, LH, oestradiol, inhibin A, B, pro-alphaC and total alpha inhibin were measured by immunoassay prior to and following gonadotrophin stimulation. Immunoactive FSH, LH and oestradiol blood concentrations following pituitary down regulation were similar in the two groups being <2.0, <3.6 IU/l and <82 pmol/l respectively. The units of FSH given (2230 versus 2764 IU; Normegon versus rFSH), duration of treatment (9.1 versus 9.4 days) and number of follicles of > or =14mm on the day of human chorionic gonadotrophin (HCG) administration (17 versus 14) were also similar. Inhibin A or B concentrations rose similarly during Normegon or rFSH administration, peaking at days 9-11. Total alpha and pro-alphaC inhibin concentrations were lower (P < 0.05) in the rFSH group during days 10 and 11 of treatment being 18.9 +/- 15.9 ng/ml (Normegon) and 4.6 +/- 2.8 ng/ml (rFSH) for total alpha inhibin and 8.5 +/- 6.8 ng/ml (Normegon) and 2.8 +/- 1.6 ng/ml (rFSH) for pro-alphaC inhibin on day 10. Overall, higher total alpha inhibin concentrations were associated with more mature follicles and oocytes, greater fertilization rates and better quality embryos. We conclude that inhibin A and B secretion was similar in both groups and is primarily controlled by FSH, whereas total alpha inhibin and pro-alphaC increased preferentially in the Normegon group over the rFSH group, indicating that they are, in part, stimulated by LH. PMID- 11387275 TI - A documented clomiphene-induced follicular development in pregnancy. AB - A case of clomiphene-induced follicular growth in pregnancy is presented in a 34 year-old woman with ectopic pregnancy treated at a university teaching hospital. Following administration of clomiphene citrate in pregnancy, follicular growth to 18 mm mean diameter was noted. It is concluded that clomiphene citrate can induce follicular growth and maturation in pregnancy, possibly by reversing pregnancy induced pituitary suppression. PMID- 11387276 TI - Maternal origin of monosomy 21 derived from ICSI. AB - With the worldwide diffusion of the intracytoplasmic sperm injection (ICSI) procedure in recent years, the issue of possible genetic risks of this new and powerful technique has attracted considerable attention. An important concern is whether ICSI facilitated the passage of genetic defects from spermatozoa to offspring. ICSI was performed with spermatozoa from a frozen-thawed sperm sample from a testicular sperm extraction (TESE) of a 38 year old man who suffered from azoospermia. His wife was 36 years old. The resulting pregnancy spontaneously aborted at 8 weeks gestation after embryo replacement. Cytogenetic investigation displayed monosomy 21. The paternal origin of the single chromosome 21 was determined by molecular analysis. The segregation error leading to loss of one chromosome 21 is likely to have occurred during oogenesis rather than as a direct consequence of ICSI. Nonetheless, monosomy 21 is extremely rare and it cannot be excluded that ICSI assisted the fertilization of an abnormal oocyte. PMID- 11387277 TI - A randomized, double-blind clinical trial using fixed daily doses of 100 or 200 IU of recombinant FSH in ICSI cycles. AB - The effect of 100 and 200 IU per day recombinant FSH (rFSH) on numbers of oocytes retrieved and the total dose used in ovarian stimulation before intracytoplasmic sperm injection was investigated in a double-blind, randomized multicentre trial. A total of 91 women was treated with a low-dose protocol and 88 with a high-dose regimen at five centres. For each started cycle, significantly more oocytes were retrieved in the 200 IU group than in 100 IU group (12.0 versus 5.7, P < 0.001); total rFSH consumption was 1121 and 1875 IU in the low- and high-dose groups respectively. Significant variations were noted between centres with regard to numbers of oocytes collected per started cycle, ranging from 2.8 to 7.2 in the 100 IU group and from 9.0 to 19.1 in the high-dose group. Exploratory analyses of secondary outcomes suggested there were no differences in vital pregnancy rates per started cycle (19.2 versus 16.9%) and per embryo transfer (26.2 versus 19.3%) in the low- and high dose groups respectively. There were four hospitalizations due to ovarian hyperstimulation syndrome, all in the 200 IU group. PMID- 11387278 TI - Semen sample collection in medium enhances the implantation rate following ICSI in patients with severe oligoasthenoteratozoospermia. AB - Ejaculation in medium increases the proportion of antibody-free spermatozoa in semen samples containing anti-sperm antibodies and thereby enhances the fertilization rate in vitro. The aim of this study was to investigate whether this technique is also beneficial in semen samples with severe oligoasthenoteratozoospermia (OAT) where bacteria and detritus are often present. A prospective randomized controlled trial was carried out to study the results of sperm preparation and fertilization and pregnancy rates after intracytoplasmic sperm injection (ICSI) for OAT. Of the 114 couples (one cycle per couple) studied between 1998 and 2000, 55 men were randomized to have semen collection into sterile dry pots (group A) and the remaining 59 had samples collected into 20 ml HEPES buffered Ham's F-10 medium with 10% human serum albumin (group B). In group B the ejaculates were incubated for 30 min and mixed gently. The samples were then processed by mild centrifugation and washing followed by a mini-swim-up technique. The ejaculates in group A were prepared by the swim-up procedure only. The overall fertilization rate was 71.8% and was similar in groups A (fertilization rate = 66.7%) and B (fertilization rate = 64.3%). In group A, 10/55 clinical pregnancies were recorded (pregnancy rate 18%), with an implantation rate (IR) of 6.9% per embryo. In group B, 16 of 59 patients conceived leading to significantly higher implantation (9.9%, P < 0.001) and clinical pregnancy rates (27%, P < 0.001). It is postulated that the addition of medium before liquefaction could inhibit the binding of bacteria and detritus to the sperm surface and may diminish DNA damage caused by reactive oxygen species, leading to improved efficiency of fertilization. The results demonstrate that the addition of HEPES buffered Ham's F-10 medium to sample collection pots significantly improves the pregnancy rate after ICSI in patients with severe OAT. PMID- 11387279 TI - Uterine contractility decreases at the time of blastocyst transfers. AB - High-frequency uterine contractions at the time of non-cavitating embryo transfer influence adversely IVF-embryo transfer outcome. This prompted us to quantify prospectively the possible decline in uterine contraction frequency occurring during later stages of the luteal phase of ovarian stimulation, up to the time of blastocyst transfers, in 43 IVF-embryo transfer candidates. Contractility was assessed on the day of human chorionic gonadotrophin (HCG) administration, 4 days after HCG (non-cavitating embryo transfer; HCG + 4), and 7 days after HCG (blastocyst transfers; HCG + 7). For this, 2 min sagittal uterine scans were obtained by ultrasound and digitized with a computerized system for the assessment of uterine contraction frequency. Our results indicated that a slight, yet significant, decrease in uterine contraction frequency, observed from the day of HCG (4.4 +/- 0.2 contractions/min) to HCG + 4 (3.5 + 0.2 contractions/min), was followed by a more pronounced, additional decrease between HCG + 4 and HCG + 7 (1.5 +/- 0.2 contractions/min; P < 0.001). In conclusion, during the luteal phase of ovarian stimulation, uterine contractility decreases progressively, and reaches a nearly quiescent status 7 days after HCG administration, at the time of blastocyst transfers. It is possible that such a uterine relaxation assists blastocyst implantation. PMID- 11387280 TI - Embryo donation: outcome and attitudes among embryo donors and recipients. AB - The outcome of an embryo donation programme was evaluated and attitudes among donors and recipients studied by means of a questionnaire survey. A total of 27 couples went through 54 treatment cycles with frozen-thawed embryos donated by other infertile couples. The indications for treatment were premature or incipient ovarian failure in combination with severe male factor infertility. The mean age of the recipient women was 36 years, and that of the recipient men was 35 years. The mean duration of infertility was 8 years (range 2-19 years). Forty six couples donated 209 excess frozen embryos to the programme. The clinical pregnancy rate in the recipients was 27.8% (15/54) per embryo transfer. An average of 1.9 embryos were transferred on each occasion. The response rate to the questionnaire was high (80-91%). Significantly more recipients (69%) than donors (47%) considered that the child should be informed about the manner of conception (P < 0.05). Some 29% of recipients and 42% of donors thought that the child should receive identifying information concerning the donor couple. The interest of the offspring, not only as regards knowing his/her genetic origin but also knowing full-blood genetic siblings, should be kept in mind in embryo donation programmes. PMID- 11387281 TI - Value of serum CA-125 concentrations as predictors of pregnancy in assisted reproduction cycles. AB - The means by which endometrial receptivity influences conception rates in assisted reproductive technology (ART) is poorly understood. As the glycoprotein CA-125 is a product of human endometrium and is measurable in the peripheral circulation, it was investigated whether it might serve as an indicator of endometrial receptivity and predictor of pregnancy. In this prospective study, serum CA-125 concentrations of 75 ART cycles were measured 1 day before and on the day of human chorionic gonadotrophin (HCG) administration, and on the day of oocyte retrieval. These women did not have endometriosis and were induced by long protocol gonadotrophin-releasing hormone analogue. Pregnancy was achieved in 35 (46.7%) cycles, but not in 40 cycles (53.3%). Serum CA-125 concentrations 1 day before and on the day of HCG administration and on the day of oocyte retrieval were significantly higher in cycles with pregnancy than in those without pregnancy (P < 0.05). It was noted that CA-125 concentrations on the day of oocyte retrieval were the best predictors of pregnancy, with concentrations >10 IU/ml having an accuracy of 86.6% for pregnancy. In conclusion, in intracytoplasmic sperm injection cycles, women with high serum CA-125 concentrations (>10 IU/ml) on the day of oocyte retrieval had very high pregnancy rates. PMID- 11387282 TI - Prophylactic salpingectomy does not impair the ovarian response in IVF treatment. AB - To examine the effect of prophylactic salpingectomy in patients with hydrosalpinges on the ovarian response to stimulation prior to IVF, 26 patients were included in a study in which they acted as their own controls. They were all part of a randomized controlled study, in which they had been randomized to no surgical intervention prior to IVF. After one or two failed cycles, they underwent laparoscopic uni- or bilateral salpingectomy of their diseased tubes. The cycles before and after surgery were compared and the ovarian response was assessed as the dose and duration of gonadotrophins and the number of retrieved and fertilized oocytes. There were no significant differences in any of the measured outcomes. The increasing age between cycles did not influence the ovarian response, assessed by a comparison with two matched control groups from the same original study; 46 patients salpingectomized before IVF and 25 patients without surgery. It is concluded that removal of hydrosalpinx as a prophylactic laparoscopic procedure does not compromise ovarian function. PMID- 11387283 TI - Elective single embryo transfer: the value of cryopreservation. AB - In-vitro fertilization is associated with a high rate of multiple pregnancies, a consequence of the number of embryos transferred. There is a challenge in avoiding even twin pregnancies in assisted reproduction, and this can be accomplished with elective single embryo transfer and a good cryopreservation programme. In our follow-up study, we analysed all our elective single embryo transfers during 1998-1999. In all these cycles at least one embryo was frozen. A total of 127 elective single embryo transfers were performed with a clinical pregnancy rate of 38.6%. The highest implantation rate was obtained with four cell embryos with <10% fragmentation (39.8%). Thirty-four patients have delivered (26.8%), one of these being a monozygotic pregnancy. In total 129 frozen-thawed cycles have been achieved in 83 patients. One frozen-thawed embryo has been transferred in 46 cycles with a clinical pregnancy rate of 17.4%, and two embryos have been transferred in 83 cycles, with a clinical pregnancy rate of 37.3%. Up until now, 66 of 125 patients in our single embryo transfer programme have delivered or have on-going pregnancies, and 77 still have embryos frozen. The cumulative delivery rate per oocyte retrieval is 52.8% and the twin rate 7.6%. We conclude that elective single embryo transfer with a good cryopreservation programme results in very acceptable pregnancy rates with a low risk of twins. This is a cost-effective practice that substantially reduces all risks associated with multiple pregnancies and lowers the cost per delivery. PMID- 11387284 TI - Feasibility of administering mifepristone as a once a month contraceptive pill. AB - Many women find the idea of a once-a-month contraceptive pill an attractive concept. Mifepristone has been shown to be effective as a contraceptive if administered in the early luteal phase. We tested the contraceptive efficacy of 200 mg of mifepristone on day luteinizing hormone (LH) + 2 in a group of 32 women who used a fertility monitor to identify the LH surge. We also recruited a control group, comprising 20 women who were trying to conceive. In this group, 12 women conceived during a total of 50 control cycles (probability of pregnancy 0.25-0.32). Women in the treatment group contributed to a total of 178 cycles and there were two pregnancies (probability of pregnancy 0.01). An LH surge was not detected in 34 cycles (19.1%). In 20 cycles (11.2%) this was due to imperfect use while 14 were monitor method failures (7.9%). Treatment with mifepristone in the early luteal phase did not disrupt the cycle length but women reported slight vaginal bleeding in 15% of the cycles. The combination of a home-use fertility monitor with once-a-month administration of mifepristone (especially if mifepristone is administered at the early luteal phase) is an acceptable contraceptive option with minimal side effects. Unfortunately, it is difficult to envisage how an easier way of defining the correct timing, which required less compliance, could be devised. PMID- 11387285 TI - Distribution of a spermicide containing Nonoxynol-9 in the vaginal canal and the upper female reproductive tract. AB - Topical, intravaginal microbicides and spermicides are greatly needed to prevent transmission of sexually transmitted diseases and/or unwanted pregnancies. The development of such compounds is a high research priority. The presumed method of action of existing, or novel, microbicides/spermicides is to provide a chemical barrier to the vaginal epithelium preventing exposure to micro-organisms. Other intravaginal products are used to treat vaginal bacteria of fungal infections. Little is known, however, about the actual or optimal initial distribution and subsequent spread of medications placed in the vagina. We describe a sensitive new technique to quantify the spread of a gel placed in the vagina using magnetic resonance imaging (MRI). Five millilitres of an over-the-counter spermicide containing Nonoxynol-9 was mixed with Gadolinium. MRI was used to quantify spread of the mixture 10 min after insertion with a standard applicator. We demonstrated contiguous spread of gel throughout the vagina. The coverage of material was thicker in the upper vagina than in the lower vagina. We also demonstrated, for the first time, that spermicidal compounds may migrate from the vaginal canal into the endocervix within 10 min of insertion. This finding suggests that topical microbicides/spermicides may act both in the vaginal canal and in the upper female genital tract. PMID- 11387286 TI - Study of the occurrence of interchromosomal effect in spermatozoa of chromosomal rearrangement carriers by fluorescence in-situ hybridization and primed in-situ labelling techniques. AB - The possibility that a chromosomal rearrangement might disturb the meiotic behaviour of chromosomes not involved in the rearrangement and favour non disjunction is a controversial issue in human cytogenetics. Using two-colour fluorescence in-situ hybridization and primed in-situ labelling techniques, we have investigated the segregation pattern of 10 chromosomes (chromosomes 1, 4, 9, 13, 15, 16, 20, 21, X and Y) in spermatozoa from nine carriers of balanced structural rearrangements and three normal men. The patients were divided into two groups according to their semen parameters. In rearrangement carriers and normal subjects, sex chromosomes and chromosome 21 displayed a higher rate of disomy than the other chromosomes. No evidence for the occurrence of interchromosomal effect was found in the spermatozoa of fertile rearrangement carriers, but significant variations were observed for all chromosomes tested in the group of infertile translocation carriers, suggesting a direct correlation between poor quality spermatozoa and increased aneuploidy rate in this group. In fertile carriers of chromosomal rearrangements, the occurrence of non-disjunction of chromosomes not involved in the rearrangement might therefore be considered as fortuitous, whereas in infertile carriers, the risk for interchromosomal effect appears to be real and should be taken into consideration in the genetic counselling of infertile couples with a male partner carrying a chromosomal rearrangement. PMID- 11387287 TI - Semen parameters, including WHO and strict criteria morphology, in a fertile and subfertile population: an effort towards standardization of in-vivo thresholds. AB - In this study, the semen analysis results of a fertile population were compared with those from a subfertile population, in order to establish normal cut-off values for the standard semen parameters with the aid of receiver operating characteristic (ROC) curve analysis. The fertile group comprised healthy males (n = 107) without any history of fertility problems, the partners of whom had had a spontaneous pregnancy within one year of unprotected intercourse and were pregnant at the time of the male's inclusion into the study. A total of 103 males from couples attending the infertility clinic, and with an initial sperm count of <20x10(6)/ml were recruited to form the subfertile population. The best discriminating parameter between the two populations was sperm morphology evaluated according to WHO criteria at a cut-off point of 31% normal spermatozoa. The other cut-off values were at 8% for the acrosome index, 45% for motility, and 4% normal spermatozoa for strict criteria. Recalculating the ROC curve cut-off values based on an assumed 50% prevalence of subfertility in an assisted reproductive setting, the cut-off points were reduced to 21% and 3% normal spermatozoa for WHO and strict criteria respectively. For motility, the new cut off value was at 20% motile spermatozoa, for motility quality at 3.5 (on a scale of 1-6), the acrosome index at 3% normal acrosomes, and the teratozoospermia index at 2.09. PMID- 11387288 TI - Aneuploidy rate in spermatozoa of selected men with abnormal semen parameters. AB - A large proportion of patients with oligoasthenoteratozoospermia (OAT) have an abnormal karyotype and hence they produce aneuploid gametes. However, a normal karyotype does not exclude the chance of having germ cell aneuploidy, since an altered intra-testicular environment not only damages spermatogenesis, but may also disrupt the mechanisms controlling chromosomal segregation during meiosis. Therefore, this study was undertaken to evaluate the rate of aneuploidy in the spermatozoa of selected patients with abnormal sperm parameters. For this purpose, sperm aneuploidy rate for chromosomes 8, 12, 18, X and Y was evaluated by multicolour fluorescence in-situ hybridization (FISH) in nine patients with teratozoospermia alone and 19 OAT patients of presumably testicular origin. Thirteen normozoospermic healthy men served as controls. Patients with teratozoospermia or OAT had significantly greater disomy and diploidy rates compared with controls, whereas the rate of nullisomy was similar. XY disomy was very low in all groups, suggesting that chromosomal non-disjunction occurs mainly during the second meiotic division. Autosome 12 disomy rate was low in both patients and controls. There was a marked variability of total sperm aneuploidy rate in both groups of patients. Sperm aneuploidy rate was negatively correlated with sperm concentration and particularly with the percentage of normal forms. In conclusion, patients with teratozoospermia or OAT have an increased rate of sperm aneuploidy. This increase is similar in both groups, suggesting that teratozoospermia may be the critical sperm parameter associated with aneuploidy. PMID- 11387289 TI - Physical activity as a possible aggravating factor for athletes with varicocele: impact on the semen profile. AB - The aim of the present study was to evaluate the influence of physical exercise on seminal parameters of male athletes with varicocele. Sixty healthy male volunteers (athletes and non-athletes, n = 30 + 30) and 60 volunteers affected by varicocele (athletes and non-athletes, n = 30 + 30) were randomly selected for a clinical study. All subjects provided at least two semen samples for routine microscopic analysis. Determinations for basal luteinizing hormone (LH), follicle stimulating hormone (FSH), prolactin, oestradiol, total and free testosterone under resting conditions were also performed. In both groups with varicocele the percentage of total and progressive forward sperm motility and the percentage of normal spermatozoa were significantly reduced. The percentage of both progressive forward motility and normal spermatozoa were significantly lower in athletes with varicocele compared with non-athletes with varicocele (P < 0.05). Only athletes with varicocele had mean left testis volume significantly lower than the contralateral testis (P < 0.05). No modifications of hormonal parameters at rest were observed in any groups. Physical activity might represent an aggravating factor for spermatogenesis in athletes with varicocele. In countries where sport eligibility is granted by an authoritative body, these results suggest the need to establish general medical criteria to guarantee the continuation of an athlete's training whilst at the same time taking care of his reproductive health. PMID- 11387290 TI - Frequency of disordered zona pellucida (ZP)-induced acrosome reaction in infertile men with normal semen analysis and normal spermatozoa-ZP binding. AB - Results of zona pellucida (ZP)-induced acrosome reaction (AR) are reported for 186 normospermic men with unexplained infertility and compared with 34 normal fertile men and 54 patients with disordered ZP-induced AR (DZPIAR) diagnosed after failure of standard IVF. For each ZP-induced AR test, four oocytes that failed to fertilize in IVF were incubated for 2 h with 2x10(6)/ml motile spermatozoa. Spermatozoa tightly bound to the ZP were recovered by aspirating the oocytes with a pipette and the AR assessed using pisum sativum agglutinin labelled with fluorescein. The standard deviation of the difference was 5.2% for repeated tests for ZP-induced AR on different ejaculates from 54 men. The ranges for the ZP-induced AR were 3-98% for normospermic infertile men, 24-95% for fertile men and 0-16% for DZPIAR patients. In the normospermic group, there was a significant correlation between ZP-induced AR and sperm concentration (Spearman r = 0.238, P < 0.001). Using ZP-induced AR < or =16% as the threshold for diagnosis of DZPIAR, the frequency of this condition in normospermic infertile men would be 25%. Thus DZPIAR is common with normospermic idiopathic infertility and this condition should be diagnosed before assisted reproductive technology since it requires intracytoplasmic sperm injection. PMID- 11387291 TI - Assessment of DNA integrity and morphology of ejaculated spermatozoa from fertile and infertile men before and after cryopreservation. AB - Cryopreservation of human spermatozoa is extensively used in artificial insemination and IVF programmes. Despite various advances in cryopreservation methodology, the recovery rate of functional post-thaw spermatozoa remains mediocre, with sperm motility being significantly decreased after freezing. This aim of this study was to investigate the effects of cryopreservation on both DNA integrity and morphology of spermatozoa from fertile and infertile men. Semen samples were obtained from 17 fertile and 40 infertile men. All samples were prepared by discontinuous Percoll density centrifugation (95.0:47.5). Samples were divided into aliquots to allow direct comparison of fresh and frozen spermatozoa from the same ejaculate. Aliquots for cryopreservation were mixed with a commercial cryoprotectant and frozen by static phase vapour cooling before plunging into liquid nitrogen. Thawing was carried out slowly at room temperature. Sperm DNA integrity was determined using a modified alkaline single cell gel electrophoresis (comet) assay and sperm morphology analysed using the Tygerberg criteria. DNA of semen and prepared spermatozoa from fertile men was found to be unaffected by cryopreservation. In marked contrast, spermatozoa from infertile men were significantly damaged by freeze-thawing. Cryopreservation had a detrimental effect on morphology of semen and prepared samples from fertile and infertile men. PMID- 11387292 TI - Evidence of reduced single-stranded testicular sperm DNA from obstructive azoospermic men after 3 days of in-vitro culture. AB - The aim of the present study was to verify whether culturing testicular tissue, to obtain a higher percentage of motile spermatozoa and a better post-thaw recovery rate, affected the ratio between single/double-stranded sperm DNA and, consequently, DNA sensitivity to damage. Testicular biopsy samples from men with obstructive and secretory azoospermia, candidates for assisted reproductive treatment, were cultured for 72 h. The percentage of motile spermatozoa and the single/double stranded DNA ratio were assessed on the day of retrieval (day 0) and again on day 3. The single/double stranded DNA ratio was measured by the acridine orange (AO) staining method. Spermatozoa were classified as green (double-stranded chromatin) or red fluorescing (single-stranded chromatin). In obstructive azoospermia, median motility was 22% (range 10-44%) on day 0 and 50% (range 38-63%) on day 3 (P < 0.01). The median percentage of red stained spermatozoa was 53.5% (range 0.1-88%) on day 0 and 20% (range 2.7-99.9%) on day 3 (P < 0.05). No changes were observed in secretory azoospermia. The culture procedure from obstructive azoospermia not only increased the post-thaw recovery rate, as previously observed, but also reduced the portion of spermatozoa containing single-stranded DNA, thereby increasing the availability of double stranded DNA spermatozoa for ICSI use. PMID- 11387293 TI - Increased aneuploidy in spermatozoa from testicular tumour patients after chemotherapy with cisplatin, etoposide and bleomycin. AB - Testicular cancer is the most common neoplasia occurring in the young male population. The PEB (cisplatin, etoposide and bleomycin) adjuvant chemotherapy usually proposed after orchidectomy in non seminomatous tumours, and in metastatic seminomas, has improved the long-term survival of these patients. Following an azoospermic period, sperm cell recovery is generally observed after treatment delivery, but little is known about the genetic consequences on these new spermatozoa. To estimate the chromosomal consequences of this chemotherapy on sperm cells during the period of recovery of spermatogenesis, sperm cell aneuploidy was studied in testicular cancer patients, at 6-18 months after PEB adjuvant chemotherapy delivery, using fluorescence in-situ hybridization (FISH) of chromosomes 7, 16, 18, X and Y with specific DNA probes. A significant increase in the frequency of diploidy and disomy for chromosomes 16, 18 and XY was observed in treated patients compared with a healthy control group. Spermatozoa aneuploidy occurring during the spermatogenesis recovery period might be a possible side effect of the PEB regimen. Thus, practitioners should be advised to provide counselling about the need for an appropriate duration of contraception. Moreover, genetic counselling should be offered in cases of pregnancy occurring soon after the end of chemotherapy. PMID- 11387294 TI - FISH assessment of aneuploidy frequencies in mature and immature human spermatozoa classified by the absence or presence of cytoplasmic retention. AB - Previously, a relationship has been found between diminished cellular maturity of human spermatozoa and low-level expression of the testis-specific chaperone protein, HspA2. Because HspA2 is a component of the synaptonemal complex in rodents, and assuming that this is also the case in men, it was postulated that the frequency of chromosomal aneuploidies would be higher in immature versus mature spermatozoa. This question was examined in spermatozoa from semen and from 80% Percoll pellets (enriched for mature spermatozoa) of the same ejaculate in 10 oligozoospermic men. Immature spermatozoa with retained cytoplasm, which signifies spermiogenetic arrest, were identified by immunocytochemistry. Using fluorescence in-situ hybridization (FISH), approximately 7000 sperm nuclei were evaluated in each of the 20 fractions (142 086 spermatozoa in all) using centromeric probes for the X, Y and 17 chromosomes. The proportions of immature spermatozoa were 45.4 +/- 3.4 versus 26.6 +/- 2.2% in the two semen versus the Percoll groups (medians: 48.2 versus 25%, P < 0.001, n = 300 spermatozoa per fraction, total 6000 spermatozoa). There was also a concomitant decline in total disomy, total diploidy and total aneuploidy frequencies in the 80% Percoll versus semen fractions (0.17 versus 0.54%, 0.14 versus 0.26% and 0.31 versus 0.81% respectively, P < 0.001 in all comparisons). The mean decline of aneuploidies was 2.7-fold. With regard to the hypothesis that aneuploidies are related to sperm immaturity, there was a close correlation between the incidence of immature spermatozoa and disomies (r = 0.7, P < 0.001) but no correlation with diploidies (r = 0.03), indicating that disomies originate primarily in immature spermatozoa. It is suggested that the common factor underlying sperm immaturity and aneuploidies is the diminished expression of HspA2. In addition, the lack of this chaperone may also cause diminished cellular transport of proteins, such as DNA repair enzymes or of the retention of cytoplasm that is extruded from normally maturing spermatozoa during spermiogenesis. PMID- 11387295 TI - Azoospermia due to testicular amyloidosis in a patient with familial Mediterranean fever. AB - We describe a patient suffering from familial Mediterranean fever (FMF) who presented to our clinic with secondary infertility of 2 years due to amyloid A amyloidosis. His spermiogram disclosed azoospermia. A testicular biopsy revealed hyalinized tubules devoid of full spermatogenesis and containing abundant amyloid, confirmed by Congo red stain. We suggest that testicular amyloidosis be taken into consideration when dealing with azoospermic FMF patients. In view of the progressive nature of amyloid accumulation in the testis we propose to follow routinely the spermiogram of FMF patients with renal amyloidosis. Furthermore, consideration of sperm cryopreservation is suggested in these cases. In FMF patients with azoospermia consideration of testicular biopsy is recommended as early as possible in order to increase the chance of sperm retrieval. PMID- 11387296 TI - Oocyte activation and Ca(2+) oscillation-inducing abilities of mouse round/elongated spermatids and the developmental capacities of embryos from spermatid injection. AB - To investigate differences in fertilization mechanisms and the potential clinical use of round/elongated spermatid, we conducted detailed studies of oocyte activation and Ca(2+) oscillation-inducing abilities in these immature sperm cells and compared these abilities against those of mature spermatozoa. When round spermatids from B(6)D(2)F(1) mice were injected, none of the oocytes was activated and no intracellular Ca(2+) ([Ca(2+)](i)) increases were observed. Elongated spermatids could induce activation normally in 87% of injected oocytes, but Ca(2+) oscillation could not be induced at all and most of the oocytes (94%) exhibited only several transient [Ca(2+)](i) rises (transient patterns). Because normal offspring could be obtained when embryos through elongated spermatid injection were transferred to foster mothers, it seems that a normal oscillation pattern of [Ca(2+)](i) is not essential for normal fertilization and embryo development. [Ca(2+)](i) patterns of injected oocytes changed from transient patterns to oscillation patterns while the injected immature sperm cells were maturing to spermatozoa. Dissociations were seen between the timing of appearance of oocyte activation and that of Ca(2+) oscillation-inducing abilities in maturing sperm cells. These dissociations may be due to differences in the thresholds to oocyte activation and Ca(2+) oscillation-inducing factor for inducing oocyte activation and Ca(2+) oscillation. PMID- 11387297 TI - Facilitated glucose transporters play a crucial role throughout mouse preimplantation embryo development. AB - The role of glucose fluctuates during preimplantation mouse embryo development, indicating that a specific interplay exists between glucose metabolism and uptake. In this study, attempts were made to characterize the role of the Na(+) coupled active and the facilitated glucose transporters (GLUT) during preimplantation development by using specific glucose analogues and transport inhibitors and by examining the expression of GLUT1. One-cell outbred mouse embryos were cultured in medium M16 (5.5 mmol/l glucose), M16 without glucose (M16-G), M16-G + 2-deoxyglucose, M16-G + 3-O-methylglucose, M16 + phlorizin and M16 + phloretin and development to the blastocyst stage assessed. The absence of glucose, or the presence of 3-O-methylglucose, which is taken up but not metabolized, did not inhibit blastocyst development. 2-Deoxyglucose, which is phosphorylated but not metabolized, inhibited blastocyst development. Culture in M16 supplemented with phlorizin, an inhibitor of Na(+)-coupled active glucose transport did not inhibit blastocyst formation. Phloretin had no effect on the cleavage of two-cell embryos to the four-cell stage, but inhibited the morula/blastocyst transition. Both phloretin and phlorizin inhibited glucose uptake in two-cell embryos. Finally, GLUT1 expression was 10-fold less in blastocysts cultured in M16 compared to in-vivo blastocysts and those cultured in M16-G. The results show that both types of glucose transporters influence preimplantation embryo development and that the embryo has an innate ability to control the uptake of glucose by regulating the expression of GLUT1. PMID- 11387298 TI - Superovulation of female mice delays embryonic and fetal development. AB - Mouse and human embryos, cultured in vitro, undergo a delay in development compared with those grown in vivo. This delay can be caused by suboptimal culture conditions, but possible influences of ovarian stimulation cannot be excluded. The objective of this study was to test the hypothesis that both in vitro and in vivo, preimplantation embryonic development and postimplantation fetal development are impaired in superovulated female mice when compared with naturally cycling controls. A delay in in-vitro blastocyst hatching and in-vivo blastocyst formation (P < 0.03 and P < 0.0001 respectively) and a 40% fetal growth retardation (P < 0.0001) were observed after superovulation in comparison with naturally cycling controls. After transfer to non-stimulated foster mothers, blastocysts from stimulated females had a lower implantation rate (P < 0.005), and developed into fewer living fetuses (P < 0.02), more resorption sites (P < 0.02) and had more pronounced growth retardation (P < 0.0001) when compared with blastocysts from naturally cycling controls. In conclusion, superovulation in the mouse causes a delayed embryonic development in vitro and in vivo, an increased abnormal blastocyst formation, a pronounced fetal growth retardation, and an increased number of resorption sites. If this observation in mice can be extrapolated to humans, it may offer an explanation for the delay in embryonic development and the low birth weight observed after IVF. PMID- 11387299 TI - A prospective randomized controlled study comparing two doses of gestodene in cyclic combined HRT preparations on endometrial physiology. AB - Postmenopausal women taking oestradiol 17-beta 2 mg daily were randomized to receive either 25 or 50 microg gestodene from day 17 to 28 of the cycle in a double-blind study. Placental protein P14 (PP14) and CA 125 concentrations in uterine flushing, endometrial morphology and irregular bleeding after 12 cycles of study were observed. Eleven and 12 women in the 25 and 50 microg groups respectively completed the study. There were no significant differences in pre treatment biochemical and morphological indices between the groups. The median PP14 concentration increased from 332 to 5800 ng/ml (P < 0.001) and from 145 to 27 160 ng/ml (P < 0.001) in the 25 and 50 microg gestodene groups respectively. No between-group significant rise of PP14 was observed. Similarly, no significant change was seen between the initial and post-treatment concentrations of CA 125 for either group. All biopsies were atrophic at inception of the study, and both regimens produced secretory endometrial transformation in the majority of biopsies. No between-group difference was observed in the morphometric indices measured, or any significant correlation between the concentrations of PP14 or CA 125 and morphology. The mean number of days of withdrawal bleeding (3.8 and 4.2 days for 25 and 50 microg respectively) were similar. In conclusion, both regimens produced a significant rise in uterine flushing concentrations of PP14, but not CA 125. PP14 is a sensitive biochemical marker in the assessment of endometrial response to hormone replacement therapy. PMID- 11387300 TI - The peritoneal fluid concentration of leptin is increased in women with peritoneal but not ovarian endometriosis. AB - This study was designed to measure leptin concentrations in the peritoneal fluid (PF) of women with different aspects of pelvic endometriosis. Among 36 consecutive women undergoing laparoscopy, nine were diagnosed as having minimal mild endometriosis (stage I-II). Among nine other subjects with advanced stage (III-IV) disease, six showed one or more ovarian endometriotic cysts as the only operative finding. The remaining 18 unaffected women constituted the control group. Patients with endometriosis had significantly higher PF leptin concentrations (32.6 +/- 16.2 versus 17.1 +/- 6.6 ng/ml, P = 0.002); this difference remained significant when corrected for body mass index (BMI) (PF leptin/BMI ratio 1.41 +/- 0.67 versus 0.76 +/- 0.28, P = 0.001). Furthermore, the PF leptin/BMI ratio was significantly higher in women with peritoneal implants than in those in whom no implant was found at laparoscopy (1.6 +/- 0.7 versus 0.83 +/- 0.33, P = 0.007). Conversely, patients with one or more ovarian endometriomata as the only finding, had a PF leptin/BMI ratio comparable with that in women where no cyst was found (1.05 +/- 0.4 versus 1.1 +/- 0.65). In women with stage I-II endometriosis, a higher mean PF leptin/BMI ratio was found compared with those affected by stage III-IV (1.78 +/- 0.68 versus 1.05 +/- 0.43, P = 0.01). These results show that during endometriosis the presence of peritoneal disease, and not of ovarian endometriotic cysts, influences leptin concentrations in PF. The data suggest that leptin may play a role in the development of peritoneal endometriosis, and that different biochemical phenomena might be involved in the pathogenesis of the ovarian form of the disease. PMID- 11387301 TI - Body composition characteristics and body fat distribution in lean women with polycystic ovary syndrome. AB - Body composition, fat distribution and bone mineral density were examined in lean women suffering from polycystic ovary syndrome (PCOS) and compared with body composition and fat distribution characteristics of weight-matched lean controls. Ten women with PCOS and a body mass index (BMI) below 25.00 (kg/m(2)) and 10 healthy women with a BMI below 25.00 (kg/m(2)) matched for age and weight and BMI as controls were enrolled in this study. Body composition and bone density were measured by dual-energy- x-ray-absorptiometry and fat distribution patterns were calculated. Although matched for age, weight and BMI, lean PCOS patients showed a significantly higher amount of body fat and lower amount of lean body mass than the controls. The majority of PCOS patients showed an intermediate or android kind of fat distribution. Only 30% of the lean PCOS patients corresponded to the definition of gynoid fat distribution while this was true of all lean controls. PMID- 11387302 TI - Conservative treatment of recurrent ovarian fibromas in a young patient affected by Gorlin syndrome. AB - The case of recurrent bilateral ovarian fibromas occurring in a 22 year old Italian girl affected by Gorlin syndrome is reported. Ovarian fibromas occur in 75% of female patients with Gorlin syndrome and their recurrence has rarely been reported in the literature. Management is guided by the benign nature of the lesion and consists of surgical removal of the fibroma. Preservation of the normal ovarian tissue is recommended even though there is risk of recurrence of the fibroma. PMID- 11387303 TI - Monozygotic twinning after assisted reproductive techniques: a phenomenon independent of micromanipulation. AB - A 3 year retrospective analysis was conducted of pregnancies achieved after various assisted reproductive treatment modalities in our infertility practice, to calculate and compare the rates of monozygotic twinning (MZT). A total of 731 pregnancies achieved after various assisted reproduction treatments were reviewed. Gonadotrophin therapy for induction of ovulation and controlled ovarian hyperstimulation (COH) yielded 129 clinical pregnancies. Conventional IVF yielded 139 pregnancies. IVF and intracytoplasmic sperm injection (ICSI) with or without assisted hatching (AH) yielded 463 pregnancies, all during the same time period. The rates of multiple pregnancy (monozygotic and dizygotic) twins and triplets were recorded. MZT was found in 1.5% of ovulation induction or COH pregnancies (2/129). The incidence of MZT after conventional IVF was 0.72% (1/139). After IVF ICSI/AH, MZT was found in 0.86% (4/463). The overall rate of MZT was 0.95% (7/731). Five cases were dizygotic triplets and two cases were monozygotic twins. We found the rate of MZT after assisted reproduction treatment increased more than two-fold over the background rate in the general population. Dizygotic triplets were found more often than monozygotic twins. The rate of MZT was consistently increased, irrespective of treatment modality or micromanipulation. This may signify that the aetiology of increased MZT after assisted reproduction is the gonadotrophin treatment rather than in-vitro conditions, micromanipulation, or multiple embryo transfer. PMID- 11387304 TI - Are elevated FSH concentrations in the pre-conceptional period a risk factor for Down's syndrome pregnancies? AB - Recent publications have reported a relation between a decreased ovarian reserve and Down's syndrome pregnancies. Using the data of a case-control study into risk factors for a Down's syndrome pregnancy, we estimated the usefulness of pre conceptional basal follicle stimulating hormone (FSH) screening (detection rate, false positive rate, positive and negative likelihood ratio, as well as the loss rate of unaffected pregnancies) to identify Down's syndrome pregnancies. The optimal detection rate of pre-conceptional basal FSH screening for Down's syndrome pregnancies was 14%, corresponding to a false positive rate of 5% and a positive likelihood ratio of 2.8. Incorporation of basal FSH screening into the regimen of first trimester serum screening followed by nuchal translucency measurement would increase the detection rate from 85 to 87%. However, basal FSH screening alone or in combination with other screening methods would cause an unacceptably high loss rate of unaffected pregnancies compared with current screening protocols, indicating that routine pre-conception basal FSH screening would not be useful to identify women at risk for a Down's syndrome pregnancy. However, when elevated basal FSH concentrations are diagnosed during subfertility evaluation, an elevated risk for a Down's syndrome pregnancy could be discussed with women who become pregnant. PMID- 11387305 TI - Subsequent pregnancy outcome in patients with spontaneous resolution of HCG after evacuation of hydatidiform mole: comparison between complete and partial mole. AB - This study compared subsequent pregnancy outcome in patients with complete and partial hydatidiform moles. Among 1052 patients with molar pregnancy (complete mole, 801; partial mole, 251) monitored at Chiba University Hospital between 1981 and 1999, 891 patients (84.7%) had spontaneous resolution of human chorionic gonadotrophin (HCG) after mole evacuation, and 161 patients (15.3%) required chemotherapy. Of the 891 patients, 438 (49.2%) had 650 subsequent pregnancies. The pregnancy outcome was not significantly different in patients with complete and partial moles, and was comparable with that in the general Japanese population. The incidence of repeat molar pregnancy in patients with complete and partial mole (1.3 and 1.5% respectively) was 5-fold higher than that of the general population, while no increased risk of persistent gestational trophoblastic tumour (GTT) associated with later molar pregnancy was observed. During HCG follow-up, 10 patients (1.1%) developed secondary high-risk GTT between 14 and 54 months after mole evacuation. The incidence of high-risk GTT in patients with and without subsequent pregnancies was 0.46% (2/438) and 1.8% (8/453) respectively (P = 0.1243). In conclusion, patients with complete and partial mole can anticipate a normal future reproductive outcome, and pregnancies after experiencing hydatidiform mole may not affect the development of high-risk GTT. PMID- 11387306 TI - High anti-paternal cytotoxic T-lymphocyte precursor frequencies in women with unexplained recurrent spontaneous abortions. AB - A number of cases of unexplained (idiopathic) recurrent spontaneous abortions may be attributable to immunological mechanisms. Several lines of evidence indicate that some immunocompetent effector cell populations play an important role in the pathogenesis of unexplained miscarriages. However a suitable method is lacking for defining an existing immunological background of recurrent spontaneous abortions. We tried to find a useful cellular immunological method, that is suitable for predicting the eventual immunological cause in the case of unexplained recurrent spontaneous abortions. We have examined the anti-paternal cytotoxic T-lymphocyte precursor frequencies by cell-mediated lympholysis and limiting dilution analysis in the peripheral blood of women with recurrent spontaneous abortions in order to reveal the functional role of this cell population in spontaneous abortions. An extremely high partner allo-antigen specific cytotoxic T-lymphocyte precursor frequency was determined in the case of all those habitual aborters, where no other than an immunological cause could be responsible for the abortions. This phenomenon supports the important role of the T-lymphocytes in this disorder. We suggest that the immunological background of recurrent spontaneous miscarriages might be determined on the basis of a very high cytotoxic T-lymphocyte precursor frequency. This diagnostic test might be useful in selecting patients for immunotherapy. PMID- 11387307 TI - Bcl-2 expression as a novel immunohistochemical marker for ruptured tubal ectopic pregnancy. AB - Programmed cell death by apoptosis occurs in fetal and maternal tissues during early pregnancy and plays an important role during implantation, decidualization, and in fetal development. In the regulation of apoptosis, bcl-2 is one of the central controlling genes, and acts by protecting the cell against apoptosis. It is postulated that invasiveness of ectopic trophoblast towards and through the muscularis zone of the tubal wall consequently leading to tubal rupture might be due to disturbed regulation of apoptosis. By means of immunohistochemistry and a computerized image analysis, bcl-2 immunostaining was localized and quantified in 36 randomly selected paraffin-embedded ectopic trophoblast tissue specimens collected from women undergoing surgery for ruptured (n = 18) and non-ruptured (n = 18) tubal ectopic pregnancies. Immunostaining was found in the villi syncytiotrophoblast in all patients, while the percentage of positive bcl-2 immunostained area (%PA) (P = 0.0009) and staining intensity (P = 0.0042) were consistently greater in the group of ruptured ectopic pregnancies. Including the variables %PA and saturation into a logistic regression model for a probability threshold of 0.5 (<0.5 = non-ruptured ectopic pregnancy, >0.5 = ruptured ectopic pregnancy) to identify tubal rupture, a sensitivity and specificity of 94.4% were found. It is suggested that elevated bcl-2 immunostaining in the syncytiotrophoblast layer reflects unlimited cell survival of ectopic trophoblast and could lead to the establishment of a circulating marker for tubal rupture. PMID- 11387308 TI - Differential expression of TGF-beta1 and TGF-beta3 in serosal tissues of human intraperitoneal organs and peritoneal adhesions. AB - Elevated local expression of transforming growth factor (TGF-beta) has been associated with increased incidence of peritoneal adhesion formation. In this study we determine whether differences in basal expression of TGF-beta in serosal tissue of peritoneal organs correlate with incidence of adhesion formation. Serosal tissue of parietal peritoneum, uterus, oviduct, ovary, omentum, large and small bowels as well as adhesions, skin, fascia, subcutaneous tissue, peritoneal fluid and serum were collected from 57 subjects with/without adhesions who were undergoing abdominal/pelvic surgery. To determine TGF-beta1 and TGF-beta3 mRNA and protein expression, total RNA and protein were isolated from these tissues and along with the fluids, subjected to quantitative RT-PCR and enzyme-linked immunosorbent assay (ELISA) respectively. Tissue sections were immunostained for TGF-beta1 and TGF-beta3 protein. We found that TGF-beta1 and TGF-beta3 mRNA and protein are expressed in these tissues and present in peritoneal fluids and serum, with considerable variations in level of their expression. Comparatively, there was more variation in TGF-beta1 than TGF-beta3 expression without age or gender relation. Adhesions express a significantly higher TGF-beta1 mRNA and have the highest TGF-beta1:TGF-beta3 ratio, with lowest concentrations and ratio detected in omentum, small and large bowels; in contrast uterus expresses higher TGF-beta3, with lowest concentrations detected in subcutaneous tissue and large bowels (P < 0.05). A similar trend was also observed for total (active + latent) TGF-beta1 protein expression, with low active TGF-beta1 that was not significantly different among the tissue extracts and fluids. However, the lowest active:total TGF-beta1 ratio was found in adhesions and ovary. In subjects with adhesions, the adhesions express significantly more TGF-beta1 compared to parietal peritoneum (P < 0.05). Immunoreactive TGF-beta1 and TGF-beta3 protein were present in various cell types in these tissues with intensity reflecting their mRNA and protein expression. In conclusion, we provided evidence that serosal tissue of various peritoneal organs and adhesions express TGF-beta1 and TGF-beta3. Since TGF-beta is expressed differently in these tissues and tissue injury often alters the expression of TGF-beta, we propose that tissues with a higher basal expression of TGF-beta may become predisposed to develop more adhesions compared to others. PMID- 11387309 TI - Guidelines for counselling in infertility: outline version. AB - The Guidelines for Counselling in Infertility describe the purpose, objectives, typical issues and communication skills involved in providing psychosocial care to individuals using fertility services. The Guidelines are presented in six sections. The first section describes how infertility consultations differ from other medical consultations in obstetrics and gynaecology, whereas the second section addresses fundamental issues in counselling, such as what is counselling in infertility, who should counsel and who is likely to need counselling. Section 3 focuses on how to integrate patient-centred care and counselling into routine medical treatment and section 4 highlights some of the special situations which can provoke the need for counselling (e.g. facing the end of treatment, sexual problems). Section 5 deals exclusively with third party reproduction and the psychosocial implications of gamete donation, surrogacy and adoption for heterosexual and gay couples and single women without partners. The final section of the Guidelines is concerned with psychosocial services that can be used to supplement counselling services in fertility clinics: written psychosocial information, telephone counselling, self-help groups and professionally facilitated group work. This paper summarizes the different sections of the Guidelines and describes how to obtain the complete text of the Guidelines for Counselling in Infertility. PMID- 11387310 TI - Comment on the debate article: embryo implantation: the Rubicon for GnRH antagonists. PMID- 11387311 TI - Y chromosome microdeletions in 'fertile' males. PMID- 11387313 TI - Akt down-regulation of p38 signaling provides a novel mechanism of vascular endothelial growth factor-mediated cytoprotection in endothelial cells. AB - Vascular endothelial growth factor (VEGF) utilizes a phosphoinositide 3-kinase (PI 3-kinase)/Akt signaling pathway to protect endothelial cells from apoptotic death. Here we show that PI 3-kinase/Akt signaling promotes endothelial cell survival by inhibiting p38 mitogen-activated protein kinase (MAPK)-dependent apoptosis. Blockade of the PI 3-kinase or Akt pathways in conjunction with serum withdrawal stimulates p38-dependent apoptosis. Blockade of PI 3-kinase/Akt also led to enhanced VEGF activation of p38 and apoptosis. In this context, the pro apoptotic effect of VEGF is attenuated by the p38 MAPK inhibitor SB203580. VEGF stimulation of endothelial cells or infection with an adenovirus expressing constitutively active Akt causes MEKK3 phosphorylation, which is associated with decreased MEKK3 kinase activity and down-regulation of MKK3/6 and p38 MAPK activation. Conversely, activation-deficient Akt decreases VEGF-stimulated MEKK3 phosphorylation and increases MKK/p38 activation. Activation of MKK3/6 is not dependent on Rac activation since dominant negative Rac does not decrease p38 activation triggered by inhibition of PI 3-kinase. Thus, cross-talk between the Akt and p38 MAPK pathways may regulate the level of cytoprotection versus apoptosis and is a new mechanism to explain the cytoprotective actions of Akt. PMID- 11387315 TI - Differential conformational requirements for activation of G proteins and the regulatory proteins arrestin and G protein-coupled receptor kinase in the G protein-coupled receptor for parathyroid hormone (PTH)/PTH-related protein. AB - After stimulation with agonist, G protein-coupled receptors (GPCRs) activate G proteins and become phosphorylated by G protein-coupled receptor kinases (GRKs), and most of them translocate cytosolic arrestin proteins to the cytoplasmic membrane. Agonist-activated GPCRs are specifically phosphorylated by GRKs and are targeted for endocytosis by arrestin proteins, suggesting a connection between GPCR conformational changes and interaction with GRKs and arrestins. Previously, we showed that by substitution of histidine for residues at the cytoplasmic side of helix 3 (H3) and helix 6 (H6) of the parathyroid hormone (PTH) receptor (PTHR), a zinc metal ion-binding site is engineered that prevents PTH-stimulated G(s) activation (Sheikh, S. P., Vilardaga, J.-P., Baranski, T. J., Lichtarge, O., Iiri, T., Meng, E. C., Nissenson, R. A., and Bourne, H. R. (1999) J. Biol. Chem. 274, 17033-17041). These data suggest that relative movements between H3 and H6 are critical for G(s) activation. Does this molecular event play a similar role in activation of GRK and arrestin and in PTHR-mediated G(q) activation? To answer this question, we utilized the two previously described mutant forms of PTHR, H401 and H402, which contain a naturally present histidine residue at position 301 in H3 and a second substituted histidine residue at positions 401 and 402 in H6, respectively. Both mutant receptors showed inhibition of PTH-stimulated inositol phosphate and cAMP generation in the presence of increasing concentrations of Zn(II). However, the mutants showed no Zn(II)-dependent impairment of phosphorylation by GRK-2. Likewise, the mutants were indistinguishable from wild-type PTHR in the ability to translocate beta arrestins/green fluorescent protein to the cell membrane and were also not affected by sensitivity to Zn(II). These results suggest that agonist-mediated phosphorylation and internalization of PTHR require conformational switches of the receptor distinct from the cAMP and inositol phosphate signaling state. Furthermore, PTHR sequestration does not appear to require G protein activation. PMID- 11387317 TI - Salmonella enteritidis FliC (flagella filament protein) induces human beta defensin-2 mRNA production by Caco-2 cells. AB - Antimicrobial peptides are crucial for host defense at mucosal surfaces. Bacterial factors responsible for induction of human beta-defensin-2 (hBD-2) mRNA expression in Caco-2 human carcinoma cells were determined. Salmonella enteritidis, Salmonella typhimurium, Salmonella typhi, Salmonella dublin, and culture supernatants of these strains induced hBD-2 mRNA expression in Caco-2 human carcinoma cells. Using luciferase as a reporter gene for a approximately 2.1-kilobase pair hBD-2 promoter, the hBD-2-inducing factor in culture supernatant of S. enteritidis was isolated. The supernatant factor was heat stable and proteinase-sensitive. After purification by anion exchange and gel filtration chromatography, the hBD-2-inducing factor was identified as a 53-kDa monomeric protein with the amino-terminal sequence AQVINTNSLSLLTQNNLNK, which is identical to that of the flagella filament structural protein (FliC) of S. enteritidis. Consistent with this finding, the 53-kDa protein reacted with anti FliC antibody, which prevented its induction of hBD-2 mRNA in Caco-2 cells. In agreement, the hBD-2-inducing activity in culture supernatant was completely neutralized by anti-FliC antibody. In gel retardation analyses, FliC increased binding of NF-kappaB (p65 homodimer) to hBD-2 gene promoter sequences. We conclude that S. enteritidis FliC induces hBD-2 expression in Caco-2 cells via NF kappaB activation and thus plays an important role in up-regulation of the innate immune response. PMID- 11387316 TI - Human bile salt export pump promoter is transactivated by the farnesoid X receptor/bile acid receptor. AB - The bile salt excretory pump (BSEP, ABCb11) is critical for ATP-dependent transport of bile acids across the hepatocyte canalicular membrane and for generation of bile acid-dependent bile secretion. Recent studies have demonstrated that the expression of this transporter is sensitive to the flux of bile acids through the hepatocyte, possibly at the level of transcription of the BSEP gene. To determine the mechanisms underlying the regulation of BSEP by bile acids, the promoter of the BSEP gene was cloned. The sequence of the promoter contained an inverted repeat (IR)-1 element (5'-GGGACA T TGATCCT-3') at base pairs -63/-50 consisting of two nuclear receptor half-sites organized as an inverted repeat and separated by a single nucleotide. This IR-1 element has been shown in several recent studies to serve as a binding site for the farnesoid X receptor (FXR), a nuclear receptor for bile acids. FXR activity requires heterodimerization with RXR alpha, and when bound by bile acids, the complex effectively regulates the transcription of several genes involved in bile acid homeostasis. Gel mobility shift assays demonstrated specific binding of FXR/RXR alpha heterodimers to the IR-1 element in the BSEP promoter. In HepG2 cells, co transfection of FXR and RXR alpha is required to attain full transactivation of the BSEP promoter by bile acids. Two FXR transactivation-deficient mutants (an AF 2 deletion and a W469A point mutant) failed to transactivate, indicating that the effect of bile acids is FXR-dependent. Further, mutational analysis confirms that the FXR/RXR alpha heterodimer activates transcription through the IR-1 site in the human BSEP promoter. These results demonstrate a mechanism by which bile acids transcriptionally regulate the activity of the bile salt excretory pump, a critical component involved in the enterohepatic circulation of bile acids. PMID- 11387318 TI - Differential effects of chronic ethanol treatment on N-methyl-D-aspartate R1 splice variants in fetal cortical neurons. AB - Functional N-methyl-D-aspartate receptors consisting of NR1 and NR2 subunits are an important site of action of ethanol. Chronic ethanol treatment increases the NR1 polypeptide levels in vivo and in vitro. Chronic ethanol treatment in vitro does not significantly alter the NR1 mRNA levels, even though under similar culture conditions ethanol (50 mm, 5 days) enhances the half-life of NR1 mRNA in fetal cortical neurons. To address this phenomenon, we determined by reverse transcription-polymerase chain reaction and Western blotting whether ethanol (50 mm, 5 days) has a splice variant-specific effect on the expression of the NR1 subunit in mouse fetal cortical neurons. This report analyzes for the first time the distribution of all NR1 splice variants in these neurons. Our data indicate the presence of NR1-3a,b and NR1-4a,b splice variants in cortical neurons. Chronic ethanol treatment significantly decreased the mRNA levels of exon 5 containing NR1 splice variants (NR1-3b and NR1-4b) (-E5/+E5 = 4.6 in untreated neurons and 6.1 in ethanol-treated neurons) and had no effect on the mRNA levels of NR1-3 (+E21/-E22) and NR1-4 (-E21/-E22) splice variants. At the polypeptide level, chronic ethanol treatment significantly reduced exon 5-containing splice variants (NR1-3b and NR1-4b). However, ethanol (50 mm, 5 days) induced a significant increase in polypeptide levels of NR1-4 (-E21/-E22), without any effect on NR1-3 (+E21/-E22) polypeptide levels. These results demonstrate that chronic ethanol treatment has a selective effect on the expression of NR1 splice variants at both the mRNA and polypeptide levels in mouse fetal cortical neurons. PMID- 11387319 TI - Binding specificity for RACK1 resides in the V5 region of beta II protein kinase C. AB - Identification of selective anchoring proteins responsible for specialized localization of specific signaling proteins has led to the identification of new inhibitors of signal transduction, inhibitors of anchoring protein-ligand interactions. RACK1, the first receptor for activated C kinase identified in our lab, is a selective anchoring protein for betaII protein kinase C (betaIIPKC). We previously found that at least part of the RACK1-binding site resides in the C2 domain of betaIIPKC (Ron, D., Luo, J., and Mochly-Rosen, D. (1995) J. Biol. Chem. 270, 24180-24187). Here we show that the V5 domain also contains part of the RACK1-binding site in betaIIPKC. In neonatal rat cardiac myocytes, the betaIIV5-3 peptide (amino acids 645-650 in betaIIPKC) selectively inhibited phorbol 12 myristate 13-acetate (PMA)-induced translocation of betaIIPKC and not betaIPKC. In addition, the betaIIV5-3 peptide inhibited cardiac myocyte hypertrophy in PMA treated cells. Interestingly, betaIV5-3 (646-651 in betaIPKC), a selective translocation inhibitor of betaIPKC, also inhibited PMA-induced cardiac myocyte hypertrophy, demonstrating that both betaI- and betaIIPKC are essential for this cardiac function. Therefore, the betaIIV5 domain contains part of the RACK1 binding site in betaIIPKC; a peptide corresponding to this site is a selective inhibitor of betaIIPKC and, hence, enables the identification of betaIIPKC selective functions. PMID- 11387320 TI - Deletion of the Src homology 3 domain and C-terminal proline-rich sequences in Bcr-Abl prevents Abl interactor 2 degradation and spontaneous cell migration and impairs leukemogenesis. AB - The hematopoietic cells from patients with Bcr-Abl-positive chronic myelogenous leukemia exhibit multiple abnormalities of cytoskeletal function. The molecular events leading to these abnormalities are not fully understood. Previously we showed that Bcr-Abl elicits ubiquitin-dependent degradation of Abl interactor proteins. Because recent studies have suggested a role of Abl interactor proteins in the pathway that regulates cytoskeletal function, we investigated whether mutations in Bcr-Abl that interfere with the signaling to Abl interactor proteins affect its leukemogenic activity. We report here that the Src homology 3 domain and C-terminal proline-rich sequences of Bcr-Abl are required for its binding to Abl interactor 2 as well as for the induction of Abl interactor 2 degradation. Although the deletion of these regions did not affect the ability of the mutant Bcr-Abl to transform hematopoietic cells to growth factor independence, it abrogated its ability to stimulate spontaneous cell migration on fibronectin coated surfaces. Furthermore, the mutant Bcr-Abl, defective in binding to Abl interactor 2 and inducing its degradation, failed to induce chronic myelogenous leukemia-like disease in mouse. These results are consistent with a role of Abl interactor proteins in the regulation of cytoskeletal function as well as in the pathogenesis of Bcr-Abl-induced leukemogenesis. PMID- 11387321 TI - Mutation in PMR1, a Ca(2+)-ATPase in Golgi, confers salt tolerance in Saccharomyces cerevisiae by inducing expression of PMR2, an Na(+)-ATPase in plasma membrane. AB - Sodium tolerance in yeast is enhanced by continuous activation of calcineurin, a Ca(2+)/calmodulin-dependent protein phosphatase that is required for modulation of the Na(+) efflux mechanism. We isolated several salt-tolerant mutations with the treatment of ethylmethane sulfonate under high salt stress. One of the mutations was mapped in the PMR1 gene. Pmr1p, the P-type Ca(2+)-ATPase in the Golgi apparatus, regulates a cytosolic Ca(2+) level in various responses. Cytosolic Ca(2+) concentration in the pmr1 mutant is highly maintained, and thus calcineurin is activated continuously. The treatment of FK506, a specific inhibitor of calcineurin, abolishes the salt-tolerant phenotype of the pmr1 mutant. Activated calcineurin induces the expression of PMR2, encoding the P-type Na(+)-ATPase, through the specific transcription factor, Tcn1p/Crz1p. Also, expression of the PMR2::lacZ reporter gene in the pmr1 mutant was higher than that in wild type. We propose that the pmr1 mutation confers salt tolerance through continuous activation of calcineurin and that Pmr1p might act as a major Ca(2+)-ATPase under high salt stress. PMID- 11387322 TI - The adapter protein apoptotic protease-activating factor-1 (Apaf-1) is proteolytically processed during apoptosis. AB - Apoptotic protease-activating factor-1 (Apaf-1), a key regulator of the mitochondrial apoptosis pathway, consists of three functional regions: (i) an N terminal caspase recruitment domain (CARD) that can bind to procaspase-9, (ii) a CED-4-like region enabling self-oligomerization, and (iii) a regulatory C terminus with WD-40 repeats masking the CARD and CED-4 region. During apoptosis, cytochrome c and dATP can relieve the inhibitory action of the WD-40 repeats and thus enable the oligomerization of Apaf-1 and the subsequent recruitment and activation of procaspase-9. Here, we report that different apoptotic stimuli induced the caspase-mediated cleavage of Apaf-1 into an 84-kDa fragment. The same Apaf-1 fragment was obtained in vitro by incubation of cell lysates with either cytochrome c/dATP or caspase-3 but not with caspase-6 or caspase-8. Apaf-1 was cleaved at the N terminus, leading to the removal of its CARD H1 helix. An additional cleavage site was located within the WD-40 repeats and enabled the oligomerization of p84 into a approximately 440-kDa Apaf-1 multimer even in the absence of cytochrome c. Due to the partial loss of its CARD, the p84 multimer was devoid of caspase-9 or other caspase activity. Thus, our data indicate that Apaf-1 cleavage causes the release of caspases from the apoptosome in the course of apoptosis. PMID- 11387323 TI - Expression of serotonin receptors in bone. AB - The 5-hydroxytryptamine (5-HT) receptors 5-HT(2A), 5-HT(2B), and 5-HT(2C) belong to a subfamily of serotonin receptors. Amino acid and mRNA sequences of these receptors have been published for several species including man. The 5-HT(2) receptors have been reported to act on nervous, muscle, and endothelial tissues. Here we report the presence of 5-HT(2B) receptor in fetal chicken bone cells. 5 HT(2B) receptor mRNA expression was demonstrated in osteocytes, osteoblasts, and periosteal fibroblasts, a population containing osteoblast precursor cells. Pharmacological studies using several agonists and antagonists showed that occupancy of the 5-HT(2B) receptor stimulates the proliferation of periosteal fibroblasts. Activity of the 5-HT(2A) receptor could however not be excluded. mRNA for both receptors was shown to be equally present in adult mouse osteoblasts. Osteocytes, which showed the highest expression of 5-HT(2B) receptor mRNA in chicken, and to a lesser extent osteoblasts, are considered to be mechanosensor cells involved in the adaptation of bone to its mechanical usage. Nitric oxide is one of the signaling molecules that is released upon mechanical stimulation of osteocytes and osteoblasts. The serotonin analog alpha-methyl-5 HT, which preferentially binds to 5-HT(2) receptors, decreased nitric oxide release by mechanically stimulated mouse osteoblasts. These results demonstrate that serotonin is involved in bone metabolism and its mechanoregulation. PMID- 11387324 TI - Crystal structure of the CheA histidine phosphotransfer domain that mediates response regulator phosphorylation in bacterial chemotaxis. AB - The x-ray crystal structure of the P1 or H domain of the Salmonella CheA protein has been solved at 2.1-A resolution. The structure is composed of an up-down up down four-helix bundle that is typical of histidine phosphotransfer or HPt domains such as Escherichia coli ArcB(C) and Saccharomyces cerevisiae Ypd1. Loop regions and additional structural features distinguish all three proteins. The CheA domain has an additional C-terminal helix that lies over the surface formed by the C and D helices. The phosphoaccepting His-48 is located at a solvent exposed position in the middle of the B helix where it is surrounded by several residues that are characteristic of other HPt domains. Mutagenesis studies indicate that conserved glutamate and lysine residues that are part of a hydrogen bond network with His-48 are essential for the ATP-dependent phosphorylation reaction but not for the phosphotransfer reaction with CheY. These results suggest that the CheA-P1 domain may serve as a good model for understanding the general function of HPt domains in complex two-component phosphorelay systems. PMID- 11387325 TI - The length, phosphorylation state, and primary structure of the RNA polymerase II carboxyl-terminal domain dictate interactions with mRNA capping enzymes. AB - The carboxyl-terminal domain (CTD) of elongating RNA polymerase II serves as a landing pad for macromolecular assemblies that regulate mRNA synthesis and processing. The capping apparatus is the first of the assemblies to act on the nascent pre-mRNA and the one for which binding of the catalytic components is most clearly dependent on CTD phosphorylation. The present study highlights a distinctive strategy of cap targeting in fission yeast whereby the triphosphatase (Pct1) and guanylyltransferase (Pce1) enzymes of the capping apparatus do not interact physically with each other (as they do in budding yeast and metazoans), but instead bind independently to the phosphorylated CTD. In vivo interactions of Pct1 and Pce1 with the CTD in a two-hybrid assay require 12 and 14 tandem repeats of the CTD heptapeptide, respectively. Pct1 and Pce1 bind in vitro to synthetic CTD peptides containing phosphoserine uniquely at position 5 or doubly at positions 2 and 5 of each of four tandem YSPTSPS repeats, but they bind weakly (Pce1) or not at all (Pct1) to a peptide containing phosphoserine at position 2. These results illustrate how remodeling of the CTD phosphorylation array might influence the recruitment and dissociation of the capping enzymes during elongation. But how does the CTD structure itself dictate interactions with the RNA processing enzymes independent of the phosphorylation state? Using CTD-Ser5 phosphopeptides containing alanine substitutions at other positions of the heptad, we define essential roles for Tyr-1 and Pro-3 (but not Thr-4 or Pro-6) in the binding of Schizosaccharomyces pombe guanylyltransferase. Tyr-1 is also essential for binding and allosteric activation of mammalian guanylyltransferase by CTD Ser5-PO4, whereas alanine mutations of Pro-3 and Pro-6 reduce the affinity for the allosteric CTD-binding site. These are the first structure-activity relationships deduced for an effector function of the phosphorylated CTD. PMID- 11387326 TI - Expression pattern and secretion of human and chicken heparanase are determined by their signal peptide sequence. AB - Cleavage of heparan sulfate (HS) proteoglycans affects the integrity and function of tissues and thereby fundamental phenomena, involving cell migration and response to changes in the extracellular microenvironment. The role of HS degrading enzymes, commonly referred to as heparanases, in normal development has not been identified. The present study focuses on cloning, expression, and properties of a chicken heparanase and its distribution in the developing chicken embryo. We have identified a chicken EST, homologous to the recently cloned human heparanase, to clone and express a functional chicken heparanase, 60% homologous to the human enzyme. The full-length chicken heparanase cDNA encodes a 60-kDa proenzyme that is processed at the N terminus into a 45-kDa highly active enzyme. The most prominent difference between the chicken and human enzymes resides in the predicted signal peptide sequence, apparently accounting for the chicken heparanase being readily secreted and localized in close proximity to the cell surface. In contrast, the human enzyme is mostly intracellular, localized in perinuclear granules. Cells transfected with a chimeric construct composed of the chicken signal peptide preceding the human heparanase exhibited cell surface localization and secretion of heparanase, similar to cells transfected with the full-length chicken enzyme. We examined the distribution pattern of the heparanase enzyme in the developing chicken embryo. Both the chicken heparanase mRNA and protein were expressed, as early as 12 h post fertilization, in cells migrating from the epiblast and forming the hypoblast layer. Later on (72 h), the enzyme is preferentially expressed in cells of the developing vascular and nervous systems. Cloning and characterization of heparanase, the first and single functional vertebrate HS-degrading enzyme, may lead to identification of other glycosaminoglycan degrading enzymes, toward elucidation of their significance in normal and pathological processes. PMID- 11387327 TI - Proteomic analysis of nucleoporin interacting proteins. AB - The Saccharomyces cerevisiae nuclear pore complex is a supramolecular assembly of 30 nucleoporins that cooperatively facilitate nucleocytoplasmic transport. Thirteen nucleoporins that contain FG peptide repeats (FG Nups) are proposed to function as stepping stones in karyopherin-mediated transport pathways. Here, protein interactions that occur at individual FG Nups were sampled using immobilized nucleoporins and yeast extracts. We find that many proteins bind to FG Nups in highly reproducible patterns. Among 135 proteins identified by mass spectrometry, most were karyopherins and nucleoporins. The PSFG nucleoporin Nup42p and the GLFG nucleoporins Nup49p, Nup57p, Nup100p, and Nup116p exhibited generic interactions with karyopherins; each bound 6--10 different karyopherin betas, including importins as well as exportins. Unexpectedly, the same Nups also captured the hexameric Nup84p complex and Nup2p. In contrast, the FXFG nucleoporins Nup1p, Nup2p, and Nup60p were more selective and captured mostly the Kap95p.Kap60p heterodimer. When the concentration of Gsp1p-GTP was elevated in the extracts to mimic the nucleoplasmic environment, the patterns of interacting proteins changed; exportins exhibited enhanced binding to FG Nups, and importins exhibited reduced binding. The results demonstrate a global role for Gsp1p-GTP on karyopherin-nucleoporin interactions and provide a rudimentary map of the routes that karyopherins take as they cross the nuclear pore complex. PMID- 11387328 TI - Protein-protein interaction and not glycosylation determines the binding selectivity of heterodimers between the calcitonin receptor-like receptor and the receptor activity-modifying proteins. AB - The receptor activity-modifying proteins (RAMPs) and the calcitonin receptor-like receptor (CRLR) are both required to generate adrenomedullin (AM) and calcitonin gene-related peptide (CGRP) receptors. A mature, fully glycosylated, form of CRLR was associated with (125)I-CGRP binding, upon co-expression of RAMP1 and CRLR. In contrast, RAMP2 and -3 promoted the expression of smaller, core-glycosylated, CRLR forms, which were linked to AM receptor pharmacology. Since core glycosylation is classically a trademark of immature proteins, we tested the hypothesis that the core-glycosylated CRLR forms the AM receptor. Although significant amounts of core-glycosylated CRLR were produced upon co-expression with RAMP2 or -3, cross-linking experiments revealed that (125)I-AM only bound to the fully glycosylated forms. Similarly, (125)I-CGRP selectively recognized the mature CRLR species upon co-expression with RAMP1, indicating that the glycosylation does not determine ligand-binding selectivity. Our results also show that the three RAMPs lie close to the peptide binding pocket within the CRLR RAMP heterodimers, since (125)I-AM and (125)I-CGRP were incorporated in RAMP2, 3, and -1, respectively. Cross-linking also stabilized the peptide-CRLR-RAMP ternary complexes, with the expected ligand selectivity, indicating that the fully processed heterodimers represent the functional receptors. Overall, the data indicate that direct protein-protein interactions dictate the pharmacological properties of the CRLR-RAMP complexes. PMID- 11387329 TI - AXOR12, a novel human G protein-coupled receptor, activated by the peptide KiSS 1. AB - A novel human G protein-coupled receptor named AXOR12, exhibiting 81% homology to the rat orphan receptor GPR54, was cloned from a human brain cDNA library. Heterologous expression of AXOR12 in mammalian cells permitted the identification of three surrogate agonist peptides, all with a common C-terminal amidated motif. High potency agonism, indicative of a cognate ligand, was evident from peptides derived from the gene KiSS-1, the expression of which prevents metastasis in melanoma cells. Quantitative reverse transcriptase-polymerase chain reaction was used to study the expression of AXOR12 and KiSS-1 in a variety of tissues. The highest levels of expression of AXOR12 mRNA were observed in brain, pituitary gland, and placenta. The highest levels of KiSS-1 gene expression were observed in placenta and brain. A polyclonal antibody raised to the C terminus of AXOR12 was generated and used to show localization of the receptor to neurons in the cerebellum, cerebral cortex, and brainstem. The biological significance of these expression patterns and the nature of the putative cognate ligand for AXOR12 are discussed. PMID- 11387331 TI - Reducing the environmental sensitivity of yellow fluorescent protein. Mechanism and applications. AB - Yellow mutants of the green fluorescent protein (YFP) are crucial constituents of genetically encoded indicators of signal transduction and fusions to monitor protein-protein interactions. However, previous YFPs show excessive pH sensitivity, chloride interference, poor photostability, or poor expression at 37 degrees C. Protein evolution in Escherichia coli has produced a new YFP named Citrine, in which the mutation Q69M confers a much lower pK(a) (5.7) than for previous YFPs, indifference to chloride, twice the photostability of previous YFPs, and much better expression at 37 degrees C and in organelles. The halide resistance is explained by a 2.2-A x-ray crystal structure of Citrine, showing that the methionine side chain fills what was once a large halide-binding cavity adjacent to the chromophore. Insertion of calmodulin within Citrine or fusion of cyan fluorescent protein, calmodulin, a calmodulin-binding peptide and Citrine has generated improved calcium indicators. These chimeras can be targeted to multiple cellular locations and have permitted the first single-cell imaging of free [Ca(2+)] in the Golgi. Citrine is superior to all previous YFPs except when pH or halide sensitivity is desired and is particularly advantageous within genetically encoded fluorescent indicators of physiological signals. PMID- 11387330 TI - Functional cloning of the proto-oncogene brain factor-1 (BF-1) as a Smad-binding antagonist of transforming growth factor-beta signaling. AB - Using the plasminogen activator inhibitor (PAI) promoter to drive the expression of a reporter gene (mouse CD2), we devised a system to clone negative regulators of the transforming growth factor-beta (TGF-beta) signaling pathway. We infected a TGF-beta-responsive cell line (MvLu1) with a retroviral cDNA library, selecting by fluorescence-activated cell sorter single cells displaying low PAI promoter activity in response to TGF-beta. Using this strategy we cloned the proto oncogene brain factor-1 (BF-1). BF-1 represses the PAI promoter in part by associating with both unphosphorylated Smad3 (in the cytoplasm) and phosphorylated Smad3 (in the nucleus), thus preventing its binding to DNA. BF-1 also associates with Smad1, -2, and -4; the Smad MH2 domain binds to BF-1, and the C-terminal segment of BF-1 is uniquely and solely required for binding to Smads. Further, BF-1 represses another TGF-beta-induced promoter (p15), it up regulates a TGF-beta-repressed promoter (Cyclin A), and it reverses the growth arrest caused by TGF-beta. Our results suggest that BF-1 is a general inhibitor of TGF-beta signaling and as such may play a key role during brain development. PMID- 11387332 TI - NF-kappa B-inducible BCL-3 expression is an autoregulatory loop controlling nuclear p50/NF-kappa B1 residence. AB - NF-kappa B is a transcription factor whose nuclear residence is controlled by I kappa B family members. In the NF-kappa B-I kappa B autoregulatory loop, activated (nuclear) Rel A.NF-kappa B1 induces the resynthesis of I kappa B alpha recapturing nuclear Rel A back into the cytoplasm within 1 h of stimulation. In contrast, NF-kappa B1 subunits redistribute more slowly into the cytoplasm (from 6 to 12 h). Here we examine the role of inducible cytoplasmic BCL-3 expression in terminating nuclear NF-kappa B1. Although BCL-3 is a nuclear protein in B lymphocytes, surprisingly, BCL-3 is primarily a cytoplasmic protein in HepG2 cells. Cytoplasmic BCL-3 abundance is induced 6-12 h after tumor necrosis factor alpha stimulation where it complexes with NF-kappa B1 homodimers. Moreover, BCL-3 mRNA and protein expression are induced by NF-kappa B-activating agents. Two observations are interpreted to indicate that bcl-3 is transactivated by NF-kappa B/Rel A: 1) expression of a dominant negative NF-kappa B inhibitor blocks tumor necrosis factor-alpha-induced BCL-3 expression and 2) expression of constitutively active Rel A is sufficient to induce BCL-3 expression. In gene transfer studies, we identify two high affinity NF-kappa B-binding sites, kappa B1 (located at -872 to -861 nucleotides) and kappa B2 (-106 to -96 nucleotides), and although both bind with high affinity to Rel A, only kappa B2 is required for NF-kappa B-dependent induction of the native BCL-3 promoter. Down-regulation of BCL-3 induction results in prolonged, enhanced NF-kappa B1 binding and increased NF-kappa B-dependent transcription. Together, these data suggest the presence of an NF-kappa B-BCL-3 autoregulatory loop important in terminating NF-kappa B1 action and that individual NF-kappa B isoforms are actively terminated through coordinate induction of inhibitory I kappa B molecules to restore cellular homeostasis. PMID- 11387333 TI - RGS12 and RGS14 GoLoco motifs are G alpha(i) interaction sites with guanine nucleotide dissociation inhibitor Activity. AB - The regulators of G-protein signaling (RGS) proteins accelerate the intrinsic guanosine triphosphatase activity of heterotrimeric G-protein alpha subunits and are thus recognized as key modulators of G-protein-coupled receptor signaling. RGS12 and RGS14 contain not only the hallmark RGS box responsible for GTPase accelerating activity but also a single G alpha(i/o)-Loco (GoLoco) motif predicted to represent a second G alpha interaction site. Here, we describe functional characterization of the GoLoco motif regions of RGS12 and RGS14. Both regions interact exclusively with G alpha(i1), G alpha(i2), and G alpha(i3) in their GDP-bound forms. In GTP gamma S binding assays, both regions exhibit guanine nucleotide dissociation inhibitor (GDI) activity, inhibiting the rate of exchange of GDP for GTP by G alpha(i1). Both regions also stabilize G alpha(i1) in its GDP-bound form, inhibiting the increase in intrinsic tryptophan fluorescence stimulated by AlF(4)(-). Our results indicate that both RGS12 and RGS14 harbor two distinctly different G alpha interaction sites: a previously recognized N-terminal RGS box possessing G alpha(i/o) GAP activity and a C terminal GoLoco region exhibiting G alpha(i) GDI activity. The presence of two, independent G alpha interaction sites suggests that RGS12 and RGS14 participate in a complex coordination of G-protein signaling beyond simple G alpha GAP activity. PMID- 11387334 TI - Visualization of ATP release in pancreatic acini in response to cholinergic stimulus. Use of fluorescent probes and confocal microscopy. AB - The energy providing substrate ATP can be released from various cells and act extracellularly to regulate the same cells or neighboring cells. However, the pathway for ATP release and the eliciting physiological stimulus are unclear. Recently, we showed that ATP activates P2X and P2Y purinergic receptors on pancreatic ducts. Thus, it was relevant to ask whether the upstream acini could be the source of releasable ATP and what the stimulus might be. We used freshly prepared rat pancreatic acini and applied conventional luminescence measurements of luciferin/luciferase reaction. As a new application of this reaction in confocal microscopy, we monitored luciferin fluorescence as a sign of ATP release by single acini. In addition we used quinacrine to mark ATP stores, which were similar to those marked with fluorescent ATP, 2'-(or-3')-O-(N-methylanthraniloyl) adenosine 5'-triphosphate, but only partially overlapping with those marked by acridine orange and LysoTracker Red. In functional studies we show that native pancreatic acini release ATP in response to various stimuli but most importantly to cholinergic stimulation, a very likely physiological stimulus in this epithelium. In a close vicinity of acini we detect about 9 microm ATP after cholinergic stimulation. Thus, ATP is poised as the paracrine mediator between pancreatic acini and ducts. PMID- 11387335 TI - Activation of I kappa b kinase by herpes simplex virus type 1. A novel target for anti-herpetic therapy. AB - Herpes simplex viruses (HSV) are ubiquitous pathogens causing a variety of diseases ranging from mild illness to severe life-threatening infections. HSV utilize cellular signaling pathways and transcription factors to promote their replication. Here we report that HSV type 1 (HSV-1) induces persistent activation of transcription factor NF-kappa B, a critical regulator of genes involved in inflammation, by activating the I kappa B kinase (IKK) in the early phase of infection. Activated NF-kappa B enhances HSV-1 gene expression. HSV-1-induced NF kappa B activation is dependent on viral early protein synthesis and is not blocked by the anti-herpetic drug acyclovir. IKK inhibition by the anti inflammatory cyclopentenone prostaglandin A(1) blocks HSV-1 gene expression and reduces virus yield by more than 3000-fold. The results identify IKK as a potential target for anti-herpetic drugs and suggest that cyclopentenone prostaglandins or their derivatives could be used in the treatment of HSV infection. PMID- 11387336 TI - Archaeal fructose-1,6-bisphosphate aldolases constitute a new family of archaeal type class I aldolase. AB - Fructose-1,6-bisphosphate (FBP) aldolase activity has been detected previously in several Archaea. However, no obvious orthologs of the bacterial and eucaryal Class I and II FBP aldolases have yet been identified in sequenced archaeal genomes. Based on a recently described novel type of bacterial aldolase, we report on the identification and molecular characterization of the first archaeal FBP aldolases. We have analyzed the FBP aldolases of two hyperthermophilic Archaea, the facultatively heterotrophic Crenarchaeon Thermoproteus tenax and the obligately heterotrophic Euryarchaeon Pyrococcus furiosus. For enzymatic studies the fba genes of T. tenax and P. furiosus were expressed in Escherichia coli. The recombinant FBP aldolases show preferred substrate specificity for FBP in the catabolic direction and exhibit metal-independent Class I FBP aldolase activity via a Schiff-base mechanism. Transcript analyses reveal that the expression of both archaeal genes is induced during sugar fermentation. Remarkably, the fbp gene of T. tenax is co-transcribed with the pfp gene that codes for the reversible PP(i)-dependent phosphofructokinase. As revealed by phylogenetic analyses, orthologs of the T. tenax and P. furiosus enzyme appear to be present in almost all sequenced archaeal genomes, as well as in some bacterial genomes, strongly suggesting that this new enzyme family represents the typical archaeal FBP aldolase. Because this new family shows no significant sequence similarity to classical Class I and II enzymes, a new name is proposed, archaeal type Class I FBP aldolases (FBP aldolase Class IA). PMID- 11387337 TI - Discordance between the binding affinity of mitogen-activated protein kinase subfamily members for MAP kinase phosphatase-2 and their ability to activate the phosphatase catalytically. AB - MKP-2 is a member of the mitogen-activated protein (MAP) kinase phosphatase family which has been suggested to play an important role in the feedback control of MAP kinase-mediated gene expression. Although MKP-2 preferentially inactivates extracellular signal-regulated kinase (ERK) and c-Jun NH(2)-terminal kinase (JNK) MAP kinase subfamilies, the mechanisms underlying its own regulation remain unclear. In this report, we have examined the MKP-2 interaction with and catalytic activation by distinct MAP kinase subfamilies. We found that the catalytic activity of MKP-2 was enhanced dramatically by ERK and JNK but was affected only minimally by p38. By contrast, p38 and ERK bound MKP-2 with comparably strong affinities, whereas JNK and MKP-2 interacted very weakly. Through site-directed mutagenesis, we defined the ERK/p38-binding site as a cluster of arginine residues in the NH(2)-terminal domain of MKP-2. Mutation of the basic motif abrogated its interaction with both ERK and p38 and severely compromised the catalytic activation of MKP-2 by these kinases. Unexpectedly, such mutations had little effect on JNK-triggered catalytic activation. Both in vitro and in vivo, wild type MKP-2 effectively inactivated ERK2 whereas MKP-2 mutants incapable of binding to ERK/p38 did not. Finally, in addition to its role as a docking site for ERK and p38, the MKP-2 basic motif plays a role in regulating its nuclear localization. Our studies provided a mechanistic explanation for the substrate preference of MKP-2 and suggest that catalytic activation of MKP-2 upon binding to its substrates is crucial for its function. PMID- 11387338 TI - Mouse CLK-1 is imported into mitochondria by an unusual process that requires a leader sequence but no membrane potential. AB - clk-1 has been identified and characterized in the nematode Caenorhabditis elegans as a gene that affects the rates, regularity, and synchrony of physiological processes. The CLK-1 protein is mitochondrial and is required for ubiquinone biosynthesis in yeast and in worms, but its biochemical function remains unclear. We have studied the expression of murine mclk1 in a variety of tissues, and we find that the pattern of mclk1 mRNA accumulation closely resembles that of mitochondrial genes involved in oxidative phosphorylation. The pattern of protein accumulation, however, is sharply distinct in some tissues; mCLK1 appears relatively enriched in the gut and depleted in the nervous tissue. We also show that mCLK1 is synthesized as a preprotein that is imported into the mitochondrial matrix, where a leader sequence is cleaved off and the protein becomes loosely associated with the inner membrane. However, in contrast to all known mitochondrial proteins that contain a cleavable pre-sequence, the import of mCLK1 does not require a mitochondrial membrane potential. PMID- 11387339 TI - Bimp1, a MAGUK family member linking protein kinase C activation to Bcl10 mediated NF-kappaB induction. AB - Bcl10 and MALT1, products of distinct chromosomal translocations in mucosa associated lymphoid tissue lymphoma, cooperate in activating NF-kappaB. Mice lacking Bcl10 demonstrate severe immunodeficiency associated with failure of lymphocytes to activate nuclear factor kappaB (NF-kappaB) in response to antigen receptor stimulation and protein kinase C activation. We characterize Bimp1, a new signaling protein that binds Bcl10 and activates NF-kappaB. Bimp1-mediated NF kappaB activation requires Bcl10 and IkappaB kinases, indicating that Bimp1 acts upstream of these mediators. Bimp1, Bcl10, and MALT1 form a ternary complex, with Bcl10 bridging the Bimp1/MALT1 interaction. A dominant negative Bimp1 mutant inhibits NF-kappaB activation by anti-CD3 ligation, phorbol ester, and protein kinase C expression. These results suggest that Bimp1 links surface receptor stimulation and protein kinase C activation to Bcl10/MALT1, thus leading to NF kappaB induction. PMID- 11387340 TI - Serum response factor cleavage by caspases 3 and 7 linked to apoptosis in human BJAB cells. AB - Apoptosis involves the cessation of cellular processes, the breakdown of intracellular organelles, and, finally, the nonphlogistic clearance of apoptotic cells from the body. Important for these events is a family of proteases, caspases, which are activated by a proteolytic cleavage cascade and drive apoptosis by targeting key proteins within the cell. Here, we demonstrate that serum response factor (SRF), a transcription factor essential for proliferative gene expression, is cleaved by caspases and that this cleavage occurs in proliferating murine fibroblasts and can be induced in the human B-cell line BJAB. We identify the two major sites at which SRF cleavage occurs as Asp(245) and Asp(254), the caspases responsible for the cleavage and generate a mutant of SRF resistant to cleavage in BJAB cells. Investigation of the physiological and functional significance of SRF cleavage reveals that it correlates with the loss of c-fos expression, whereby neither SRF cleavage fragment retains transcriptional activity. Moreover, the expression of a noncleavable SRF in BJAB cells suppresses apoptosis induced by Fas cross-linking. These results suggest that for apoptosis to proceed, the transcriptional events promoting cell survival and proliferation, in which SRF is involved, must first be inactivated. PMID- 11387341 TI - Marked stepwise differences within a common kinetic mechanism characterize TATA binding protein interactions with two consensus promoters. AB - Binding of the TATA-binding protein (TBP) to promoter DNA bearing the TATA sequence is an obligatory initial step in RNA polymerase II transcription initiation. The interactions of Saccharomyces cerevisiae TBP with the E4 (TATATATA) and adenovirus major late (TATAAAAG) promoters have been modeled via global analysis of kinetic and thermodynamic data obtained using fluorescence resonance energy transfer. A linear two-intermediate kinetic mechanism describes the reaction of both of these consensus strong promoters with TBP. Qualitative features common to both interactions include tightly bound TBP-DNA complexes with similar solution geometries, simultaneous DNA binding and bending, and the presence of intermediate TBP-DNA conformers at high mole fraction throughout most of the reaction and at equilibrium. Despite very similar energetic changes overall, the stepwise entropic and enthalpic compensations along the two pathways differ markedly following the initial binding/bending event. Furthermore, TBP-E4 dissociation ensues from both replacement and displacement processes, in contrast to replacement alone for TBP-adenovirus major late promoter. A model is proposed that explicitly correlates these similarities and differences with the sequence specific structural properties inherent to each promoter. This detailed mechanistic comparison of two strong promoters interacting with TBP provides a foundation for subsequent comparison between consensus and variant promoter sequences reacting with TBP. PMID- 11387342 TI - Identification of proximate regions in a complex of retinal guanylyl cyclase 1 and guanylyl cyclase-activating protein-1 by a novel mass spectrometry-based method. AB - A key challenge in studying protein/protein interactions is to accurately identify contact surfaces, i.e. regions of two proteins that are in direct physical contact. Aside from x-ray crystallography and NMR spectroscopy few methods are available that address this problem. Although x-ray crystallography often provides detailed information about contact surfaces, it is limited to situations when a co-crystal of proteins is available. NMR circumvents this requirement but is limited to small protein complexes. Other methods, for instance protection from proteolysis, are less direct and therefore less informative. Here we describe a new method that identifies candidate contact surfaces in protein complexes. The complexes are first stabilized by cross linking. They are then digested with a protease, and the cross-linked fragments are analyzed by mass spectrometry. We applied this method, referred to as COSUMAS (contact surfaces by mass spectrometry), to two proteins, retinal guanylyl cyclase 1 (RetGC1) and guanylyl cyclase-activating protein-1 (GCAP-1), that regulate cGMP synthesis in photoreceptors. Two regions in GCAP-1 and three in RetGC1 were identified as possible contact sites. The two regions of RetGC1 that are in the vicinities of Cys(741) and Cys(780) map to a kinase homology domain in RetGC1. Their identities as contact sites were independently evaluated by peptide inhibition analysis. Peptides with sequences from these regions block GCAP-1 mediated regulation of guanylyl cyclase at both high and low Ca2+ concentrations. The two regions of GCAP-1 cross-linked to these peptides were in the vicinities of Cys(17) and Cys(105) of GCAP-1. Peptides with sequences derived from these regions inhibit guanylyl cyclase activity directly. These results support a model in which GCAP-1 binds constitutively to RetGC1 and regulates cyclase activity by structural changes caused by the binding or dissociation of Ca2+. PMID- 11387343 TI - Overexpression of macrophage colony-stimulating factor receptor on microglial cells induces an inflammatory response. AB - Microglia are important in the inflammatory response in Alzheimer's disease (AD). We showed previously that macrophage colony-stimulating factor receptor (M-CSFR), encoded by the c-fms protooncogene, is overexpressed on microglia surrounding amyloid beta (Abeta) deposits in the APP(V717F) mouse model for AD. The M-CSFR is also increased on microglia after experimental brain injury and in AD. To determine the relevance of these findings, we transiently expressed M-CSFR on murine BV-2 and human SV-A3 microglial cell lines using an SV40-promoted c-fms construct. M-CSFR overexpression resulted in microglial proliferation and increased expression of inducible nitric-oxide synthase, the proinflammatory cytokines interleukin-1alpha, macrophage inflammatory protein 1-alpha, and interleukin-6 and of macrophage colony-stimulating factor (M-CSF) itself. Antibody neutralization of M-CSF showed that the M-CSFR-induced proinflammatory response was dependent on M-CSF in the culture media. By using a co-culture of c fms-transfected murine microglia and rat organotypic hippocampal slices and a species-specific real time reverse transcriptase-polymerase chain reaction assay and enzyme-linked immunosorbent assay, we showed that M-CSFR overexpression on exogenous microglia induced expression of interleukin-1alpha by the organotypic culture. These results show that increased M-CSFR expression induces microglial proliferation, cytokine expression, and a paracrine inflammatory response, suggesting that in APP(V717F) mice increased M-CSFR on microglia could be an important factor in Abeta-induced inflammatory response. PMID- 11387344 TI - An essential GTPase, der, containing double GTP-binding domains from Escherichia coli and Thermotoga maritima. AB - A gene encoding a putative GTPase containing two tandemly repeated GTP-binding domains from a hyperthermophilic bacterium, Thermotoga maritima, was cloned and expressed in Escherichia coli. The gene (TM1446) termed der is highly conserved in Eubacteria including E. coli. The purified der product (Tm-Der) has GTPase activity but no ATPase activity. GTP, GDP, and dGTP but not GMP, ATP, CTP, and UTP compete for GTP binding to Tm-Der. An optimal condition for the GTPase assay was determined to be pH 7.5 in 400 mm KCl and 5 mm MgCl(2) at 70 degrees C, where K(m), V(max), and k(cat) values were determined to be 110 microm, 3.46 microm/min, and 0.87 min(-1), respectively. A der deletion strain of E. coli was constructed by replacing the der gene (originally annotated yfgK) with a kanamycin resistance gene. The deletion strain was found to form colonies only if the cells harbored a plasmid containing der, indicating that der is essential for E. coli growth. PMID- 11387345 TI - Two splice variants of protein kinase B gamma have different regulatory capacity depending on the presence or absence of the regulatory phosphorylation site serine 472 in the carboxyl-terminal hydrophobic domain. AB - We have reported previously the cloning and characterization of human and mouse protein kinase B gamma (PKB gamma), the third member of the PKB family of second messenger-regulated serine/threonine kinases (Brodbeck, D., Cron, P., and Hemmings, B. A. (1999) J. Biol. Chem. 274, 9133--9136). Here we report the isolation of human and mouse PKB gamma 1, a splice variant lacking the second regulatory phosphorylation site Ser-472 in the hydrophobic C-terminal domain. Expression of PKB gamma 1 is low compared with PKB gamma, and it is regulated in different human tissues. We show that PKB gamma and PKB gamma 1 differ in their response to stimulation by insulin, pervanadate, peroxide, or okadaic acid. Activation of PKB gamma 1 requires phosphorylation at a single regulatory site Thr-305. Interestingly, this site is phosphorylated to a higher extent in PKB gamma compared with PKB gamma 1 upon maximal stimulation by pervanadate, and this is reflected in the respective specific kinase activities. Furthermore, upon insulin stimulation of transfected cells, PKB gamma 1 translocates to the plasma membrane to a lesser extent than PKB gamma. Taken together, these results suggest that phosphorylation of the hydrophobic motif at the extreme C terminus of PKB gamma may facilitate translocation of the kinase to the membrane and/or its phosphorylation on the activation loop site by phosphoinositide-dependent protein kinase-1. PMID- 11387346 TI - High-dose chemotherapy for breast cancer: "how do you know?". PMID- 11387347 TI - An on-site audit of the South African trial of high-dose chemotherapy for metastatic breast cancer and associated publications. AB - PURPOSE: The randomized study reported by Bezwoda et al of high-dose chemotherapy (HDC) for treatment of metastatic breast cancer was audited on site to verify the study results. Additional published studies were reviewed to determine whether they had been subject to the required institutional oversight. PATIENTS AND METHODS: Ninety patients were reported to have been randomized and treated on this trial. A log of the names, hospital numbers, entry dates, and regimen received had been provided by the principal investigator. A search of more than 15,000 sets of medical records available from two Johannesburg hospitals was performed to locate records for as many of these 90 patients as possible. Standard auditing techniques were used. Additional clinical trials published by Bezwoda were compared against the minutes of the University of the Witwatersrand Committee for Research on Human Subjects to verify review and approval. RESULTS: Records for only 61 of the 90 patients could be found. Of these 61, only 27 had sufficient records to verify eligibility for the trial by the published criteria. Of these 27, 18 did not meet one or more eligibility criteria. Only 25 patients appeared to have received their assigned therapy temporally associated with their enrollment date, and all but three of these 25 received HDC. The treatment details of individual patients were at great variance from the published data. Nine other trials reported by Bezwoda were not reviewed or approved by the appropriate institutional committee despite statements to the contrary in the publications. CONCLUSION: The multiple publications of this study do not report verifiable data, and nine other publications coauthored by the principal investigator contain at least one major untrue statement. PMID- 11387348 TI - Thymidine kinase as a proliferative marker: clinical relevance in 1,692 primary breast cancer patients. AB - PURPOSE: To assess the prognostic value of thymidine kinase (TK), an enzyme involved in the DNA synthesis salvage pathway, relative to other prognostic factors in primary breast cancer. PATIENTS AND METHODS: This retrospective study involved 1,692 patients with operable breast cancer treated in six institutions (median follow-up, 82 months). Among the 857 node-negative patients, 135 received adjuvant chemotherapy (fluorouracil, doxorubicin, cyclophosphamide [FAC] or fluorouracil, etoposide, and cisplatin [FEC]). TK was assayed in cytosol with a quantitative radioenzymatic technique. Disease-specific survival (DSS), local recurrence-free interval (LRI), and distant-relapse-free interval (DRI) were investigated. RESULTS: High TK levels were associated with large tumor size, high histologic grade, and steroid hormone receptor negativity. Univariate analysis of the entire data set showed that high TK levels were related to shorter DSS (P < 10(-5)), LRI (P < 10(-3)), and DRI (P < 10(-5)). In time-dependent Cox models, high TK levels remained an independent predictor of the three outcomes, both in the overall population and in node-negative patients, although its prognostic value decreased over time. In node-negative patients, the introduction of an interaction term in multivariate analysis suggested that chemotherapy was more efficacious for patients who had tumors with high TK contents. In node-positive patients, high TK levels were related only to an increased risk of LRI. CONCLUSION: High TK values are an important risk factor in node-negative patients and seem to be associated with a beneficial effect of adjuvant FAC or FEC in patients who received adjuvant chemotherapy. The rationale of chemotherapy for patients with slowly proliferating tumors has to be discussed from a risk-benefit point of view. PMID- 11387350 TI - Metabolic flare: indicator of hormone responsiveness in advanced breast cancer. AB - PURPOSE: The purpose of this study was to investigate whether positron emission tomography (PET) with the glucose analog [(18)F]fluorodeoxyglucose (FDG) and the estrogen analog 16 alpha-[(18)F]fluoroestradiol-17 beta (FES), performed before and after treatment with tamoxifen, could be used to detect hormone-induced changes in tumor metabolism (metabolic flare) and changes in available levels of estrogen receptor (ER). In addition, we investigated whether these PET findings would predict hormonally responsive breast cancer. PATIENTS AND METHODS: Forty women with biopsy-proved advanced ER-positive (ER(+)) breast cancer underwent PET with FDG and FES before and 7 to 10 days after initiation of tamoxifen therapy; 70 lesions were evaluated. Tumor FDG and FES uptake were assessed semiquantitatively by the standardized uptake value (SUV) method. The PET results were correlated with response to hormonal therapy. RESULTS: In the responders, the tumor FDG uptake increased after tamoxifen by 28.4% +/- 23.3% (mean +/- SD); only five of these patients had evidence of a clinical flare reaction. In nonresponders, there was no significant change in tumor FDG uptake from baseline (mean change, 10.1% +/- 16.2%; P =.0002 v responders). Lesions of responders had higher baseline FES uptake (SUV, 4.3 +/- 2.4) than those of nonresponders (SUV, 1.8 +/- 1.3; P =.0007). All patients had evidence of blockade of the tumor ERs 7 to 10 days after initiation of tamoxifen therapy; however, the degree of ER blockade was greater in the responders (mean percentage decrease, 54.8% +/- 14.2%) than in the nonresponders (mean percentage decrease, 19.4% +/- 17.3%; P =.0003). CONCLUSION: The functional status of tumor ERs can be characterized in vivo by PET with FDG and FES. The results of PET are predictive of responsiveness to tamoxifen therapy in patients with advanced ER(+) breast cancer. PMID- 11387349 TI - Effect of endocrine treatment on sexuality in premenopausal breast cancer patients: a prospective randomized study. AB - PURPOSE: To study the sexual effects of the 2-year adjuvant goserelin (Zoladex [Zeneca AB, Sodertalje, Sweden]) alone, tamoxifen alone, and Zoladex and tamoxifen in combination (ZT) versus no adjuvant endocrine therapy among premenopausal breast cancer patients with or without chemotherapy in a controlled clinical trial (a European multicenter trial: Zoladex in Premenopausal Breast Cancer Patients). PATIENTS AND METHODS: This prospective study examined several aspects of sexuality through the use of self-administered questionnaires, which were completed by patients at seven points of assessment for 3 years after randomization. RESULTS: Patients treated with chemotherapy had a higher level of sexual dysfunction than did patients who received no systemic treatment. The addition of endocrine treatment did not alter this result. In contrast, among patients who did not receive chemotherapy, Zoladex and ZT produced a significantly higher level of dysfunction from 1 to 2 years after inclusion, as compared with those who received no endocrine treatment. Tamoxifen alone did not produce side effects. After termination of endocrine treatment, sexual dysfunction began to diminish. Those with chemotherapy had high and frequently increasing levels of dysfunction even after 2 to 3 years of independent of endocrine treatment. Zoladex had a negative effect on sexual fear, which was reduced by the addition of tamoxifen. CONCLUSION: Zoladex increased sexual dysfunction during treatment among patients without chemotherapy, but the disturbances of sexual functioning were reversible. The use of adjuvant chemotherapy was associated with continued sexual problems, even at 3 years after randomization. PMID- 11387351 TI - Experience with 2-chlorodeoxyadenosine in previously untreated children with newly diagnosed acute myeloid leukemia and myelodysplastic diseases. AB - PURPOSE: To develop more effective chemotherapy regimens for childhood acute myelogenous leukemia (AML). PATIENTS AND METHODS: Between June 1991 and December 1996, we administered the nucleoside analog 2-chlorodeoxyadenosine (2-CDA) to 73 children with primary AML and 20 children with secondary AML or myelodysplastic syndrome (MDS). Patients received one or two 5-day courses of 2-CDA (8.9 mg/m(2)/d) given by continuous infusion. All patients then received one to three courses of daunomycin, cytarabine, and etoposide (DAV) remission induction therapy. RESULTS: Seventy-two patients with primary AML were assessable for response. Their rate of complete remission (CR) was 24% after one course of 2 CDA, 40% after two courses of 2-CDA, and 78% after DAV therapy. Of the 57 patients who entered CR, 11 subsequently underwent allogeneic bone marrow transplantation (BMT), and 40 underwent autologous BMT. Twenty-nine patients remain in continuous CR after BMT. Two patients remain in CR after chemotherapy only. The 5-year event-free survival (EFS) estimate was 40% (SE = 0.080%). Patients with French-American-British (FAB) M5 AML had a higher rate of CR after treatment with 2-CDA (45% after one course and 70.6% after two courses) than did others (P =.002). In contrast, no patient with FAB M7 AML (n = 10) entered CR after treatment with 2-CDA. Similarly, no patient with primary MDS (n = 6) responded to 2-CDA. Seven patients with secondary AML or MDS (n = 14) had a partial response to one course of 2-CDA. CONCLUSION: This agent was well tolerated, and its toxicity was acceptable. Future trials should examine the effectiveness of 2-CDA given in combination with other agents effective against AML. PMID- 11387353 TI - Hyperfractionated low-dose radiotherapy for high-risk neuroblastoma after intensive chemotherapy and surgery. AB - PURPOSE: To assess prognostic factors for local control in high-risk neuroblastoma patients treated with hyperfractionated 21-Gy total dose to consolidate remission achieved by dose-intensive chemotherapy and surgery. PATIENTS AND METHODS: Patients with high-risk neuroblastoma in first remission received local radiotherapy (RT) totaling 21 Gy in twice-daily 1.5-Gy fractions. RT to the primary site followed dose-intensive chemotherapy and tumor resection; the target field encompassed the extent of tumor at diagnosis, plus 3-cm margins and regional lymph nodes. RT to distant sites followed radiologic evidence of response. Local failure was correlated with clinical factors (including other consolidative treatments) and biologic findings. RESULTS: Of 99 consecutively irradiated patients followed for a median of 21.1 months from RT, 10 relapsed in or at margins of RT fields at 1 to 27 months (median, 14 months). At 36 months after RT, the probability of primary-site failure was 10.1% +/- 5.3%. No primary site relapses occurred among the 23 patients whose tumors were excised at diagnosis, but there were three such relapses among the seven patients who were irradiated with evidence of residual disease in the primary site. Four of 18 patients with MYCN-amplified disease and serum lactate dehydrogenase greater than 1,500 U/L had local failures (23.4% +/- 10.7% risk at 18 months). Acute radiotoxicities were insignificant, but three of 35 patients followed for > or = 36 months had short stature from decreased growth of irradiated vertebra. CONCLUSION: Hyperfractionated 21-Gy RT is well tolerated and, together with dose intensive chemotherapy and surgery, may help in local control of high-risk neuroblastoma. Extending the RT field to definitively encompass regional nodal groups may improve results. Visible residual disease may warrant higher RT dosing. Patients with biologically unfavorable disease may be at increased risk for local failure. RT to the primary site may not be necessary when tumors are excised at diagnosis. PMID- 11387352 TI - High-dose melphalan, etoposide, total-body irradiation, and autologous stem-cell reconstitution as consolidation therapy for high-risk Ewing's sarcoma does not improve prognosis. AB - PURPOSE: To determine whether consolidation therapy with high-dose melphalan, etoposide, and total-body irradiation (TBI) with autologous stem-cell support would improve the prognosis for patients with newly diagnosed metastatic Ewing's sarcoma (ES). PATIENTS AND METHODS: Thirty-two eligible patients with newly diagnosed ES metastatic to bone and/or bone marrow were enrolled onto this study. Treatment was initially comprised of five cycles of induction chemotherapy (cyclophosphamide, doxorubicin, and vincristine alternating with ifosfamide and etoposide) and local control. Peripheral-blood stem-cell collection was performed after the second cycle of chemotherapy, with delay if the bone marrow was persistently involved. If patients had a good response to initial therapy, they proceeded to consolidation therapy with melphalan, etoposide, TBI, and stem-cell support. RESULTS: Of the 32 eligible patients, 23 proceeded to high-dose therapy consolidation. Of the nine patients who did not proceed to consolidation, four were secondary to progressive disease and two were secondary to toxicity. Three patients died from toxicity during the high-dose phase of the therapy. The majority of the patients who underwent high-dose consolidation therapy experienced relapse and died with progressive disease. Two-year event-free survival (EFS) for all eligible patients is 20%. The 2-year post-stem-cell reconstitution EFS for the subset of 23 patients who received consolidation therapy is 24%. Analysis of peripheral-blood stem-cell collections by molecular techniques for minimal residual disease showed contamination of at least some samples by tumor cells in all three patients with available data. CONCLUSION: Consolidation with high-dose melphalan, etoposide, TBI, and autologous stem-cell support failed to improve the probability of EFS in this cohort of patients with newly diagnosed metastatic ES. PMID- 11387354 TI - Mutations and allelic loss of p53 in primary tumor DNA from potentially cured patients with colorectal carcinoma. AB - PURPOSE: To compare p53 alterations in survivors and nonsurvivors after surgery for colorectal cancer. PATIENTS AND METHODS: Twenty-nine potentially cured patients with colorectal carcinoma, without recurrent disease for more than 6 years after their primary surgery, were selected to match a group of 41 colorectal cancer patients with early metastatic spread to the liver. All patients were screened for mutations in the p53 gene, exons 5 to 9, by denaturing gradient gel electrophoresis and subsequent sequencing. RESULTS: The frequency of p53 mutations was significantly different in cured patients (60%) compared with patients with early relapse (41%, P <.05). A significant difference was found in the distribution of mutations, indicating that potentially cured patients had a different proportion of mutations in conserved regions of p53 (P =.02). This difference was explained by a significantly different frequency of mutations in exon 8 (40% v 15%, P =.03), which is part of the conserved region V. All mutations in region V were codon 273 mutations in cured patients, whereas three of four mutations were located in codon 273 in patients with metastatic disease. Allelic loss of p53 (loss of heterozygosity [LOH]) was demonstrated in 26% of the cured patients and in 39% of patients with metastatic disease (P =.36). The combination of mutation and LOH of p53 was the same (17%) in both groups. CONCLUSION: A large number of p53 mutations in colorectal cancer do not promote disease progression. Some mutations, particularly within conserved regions, may even counteract negative functional effects of other p53 structural alterations. A complete loss of p53 function was not related to survival or progression after curative operation of colorectal carcinoma. PMID- 11387355 TI - Discordance between K-ras mutations in bone marrow micrometastases and the primary tumor in colorectal cancer. AB - PURPOSE: To study bone marrow micrometastases from colorectal cancer patients for the presence of K-ras mutations and to compare their genotype with that of the corresponding primary tumor. PATIENTS AND METHODS: Bilateral iliac crest aspiration was performed in 51 patients undergoing surgery for colorectal cancer, and bone marrow micrometastases were detected by immunohistochemistry. The presence of K-ras mutations was determined by single-strand conformation polymorphism analysis on both primary tumors and paired bone marrow samples and was confirmed by sequencing. RESULTS: In six patients with primary tumor mutations, it was possible to amplify a mutated K-ras gene also from the bone marrow sample. In three of those patients the pattern of K-ras mutations differed between both samples, in two patients the mutation was identical between the bone marrow and its primary tumor, and in one patient the same mutation plus a different one were found. Fifteen of 17 K-ras mutations found in primary tumors were located in codon 12, whereas in bone marrow, five of seven mutations were found in codon 13 (P =.003). CONCLUSION: Our results demonstrate that, at least for K-ras mutations, disseminated epithelial cells are not always clonal with the primary tumor and they question the malignant genotype of bone marrow micrometastases. They also indicate that different tumoral clones may be circulating simultaneously or sequentially in the same patient. Analysis of the type of mutations suggests that cell dissemination might be an early event in colorectal carcinogenesis. PMID- 11387356 TI - Psychosocial characteristics of individuals with non-stage IV melanoma. AB - PURPOSE: Melanoma is the fastest growing solid tumor among men and women and accounts for 79% of skin cancer-related deaths. Research has identified that distress is frequently associated with a diagnosis of cancer and may slow treatment-seeking and recovery, increasing morbidity and even mortality through faster disease course. Given that the 5-year survival rates for individuals with melanoma are determined primarily by the depth and extent of spread, distress that interferes with seeking treatment has the potential to be life-threatening. PATIENTS AND METHODS: The current study was designed to identify levels of distress present in individuals seeking treatment at a large, Midwestern, multidisciplinary melanoma clinic. It also focused on determining the quality of life, level of anxiety, and coping strategies used by individuals with melanoma before treatment. Given that the course of treatment and outcome for patients with stage IV disease is vastly different from that of patients with stages I to III disease, they were excluded from the study. RESULTS: Results indicated that most individuals who are presenting to a melanoma clinic do not report a clinically significant level of distress. However, there is some variability in this, with 29% of patients reporting moderate to high levels of distress. Moreover, analyses suggest that distressed individuals are more likely to use maladaptive coping strategies, such as escape-avoidance coping, and to have poorer quality of life. CONCLUSION: Although most individuals do not present with significant levels of distress, a significant minority are distressed and rely more heavily on coping strategies that do not benefit them. Such individuals would likely benefit most from psychological intervention. PMID- 11387357 TI - Sentinel lymph node biopsy for melanoma: controversy despite widespread agreement. AB - Although sentinel lymph node (SLN) biopsy for melanoma has been adopted throughout the United States and abroad as a standard method of determining the pathologic status of the regional lymph nodes, some controversy still exists regarding the validity and utility of this procedure. SLN biopsy is a minimally invasive procedure, performed on an outpatient basis at the time of wide local excision of the melanoma, with little morbidity. Numerous studies have documented the accuracy of this procedure for identifying nodal metastases. There are four major reasons to perform SLN biopsy. First, SLN biopsy improves the accuracy of staging and provides valuable prognostic information for patients and physicians to guide subsequent treatment decisions. Second, SLN biopsy facilitates early therapeutic lymph node dissection for those patients with nodal metastases. Third, SLN biopsy identifies patients who are candidates for adjuvant therapy with interferon alfa-2b. Fourth, SLN biopsy identifies homogeneous patient populations for entry onto clinical trials of novel adjuvant therapy agents. Overall, the benefit of accurate nodal staging obtained by SLN biopsy far outweighs the risks and has important implications for patient management. PMID- 11387358 TI - Preoperative plasma levels of transforming growth factor beta(1) (TGF-beta(1)) strongly predict progression in patients undergoing radical prostatectomy. AB - PURPOSE: Elevated local and circulating levels of transforming growth factor beta(1) (TGF-beta(1)) have been associated with prostate cancer invasion and metastasis. We tested the hypothesis that preoperative plasma TGF-beta(1) levels would independently predict cancer stage and prognosis in patients who undergo radical prostatectomy. PATIENTS AND METHODS: The study group consisted of 120 consecutive patients who underwent radical prostatectomy for clinically localized prostate cancer (median follow-up, 53.8 months). Preoperative plasma levels of TGF-beta(1) were measured and correlated with pathologic parameters and clinical outcomes. TGF-beta(1) levels also were measured in 44 healthy men without cancer, in 19 men with prostate cancer metastatic to regional lymph nodes, and in 10 men with prostate cancer metastatic to bone. RESULTS: Plasma TGF-beta(1) levels in patients with lymph node metastases (14.2 +/- 2.6 ng/mL) and bone metastases (15.5 +/- 2.4 ng/mL) were higher than those in radical prostatectomy patients (5.2 +/- 1.3 ng/mL) and healthy subjects (4.5 +/- 1.2 ng/mL) (P <.001). In a preoperative analysis, preoperative plasma TGF-beta(1) level and biopsy Gleason sum both were predictors of organ-confined disease (P =.006 and P =.006, respectively) and PSA progression (P <.001 and P =.021, respectively). In a postoperative multivariate analysis, preoperative plasma TGF-beta(1) level, pathologic Gleason sum, and surgical margin status were predictors of PSA progression (P =.020,P =.020, and P =.022, respectively). In patients who progressed, preoperative plasma TGF-beta(1) levels were higher in those with presumed distant compared with local-only failure (P =.019). CONCLUSION: Plasma TGF-beta(1) levels are markedly elevated in men with prostate cancer metastatic to regional lymph nodes and bone. In men without clinical or pathologic evidence of metastases, the preoperative plasma TGF-beta(1) level is a strong predictor of biochemical progression after surgery, presumably because of an association with occult metastatic disease present at the time of radical prostatectomy. PMID- 11387359 TI - Effects of epoetin alfa on hematologic parameters and quality of life in cancer patients receiving nonplatinum chemotherapy: results of a randomized, double blind, placebo-controlled trial. AB - PURPOSE: This randomized, double-blind, placebo-controlled clinical trial assessed the effects of epoetin alfa on transfusion requirements, hematopoietic parameters, quality of life (QOL), and safety in anemic cancer patients receiving nonplatinum chemotherapy. The study also explored a possible relationship between increased hemoglobin and survival. PATIENTS AND METHODS: Three hundred seventy five patients with solid or nonmyeloid hematologic malignancies and hemoglobin levels < or = 10.5 g/dL, or greater than 10.5 g/dL but < or = 12.0 g/dL after a hemoglobin decrease of > or = 1.5 g/dL per cycle since starting chemotherapy, were randomized 2:1 to epoetin alfa 150 to 300 IU/kg (n = 251) or placebo (n = 124) three times per week subcutaneously for 12 to 24 weeks. The primary end point was proportion of patients transfused; secondary end points were change in hemoglobin and QOL. The protocol was amended before unblinding to prospectively collect and assess survival data 12 months after the last patient completed the study. RESULTS: Epoetin alfa, compared with placebo, significantly decreased transfusion requirements (P =.0057) and increased hemoglobin (P <.001). Improvement of all primary cancer- and anemia-specific QOL domains, including energy level, ability to do daily activities, and fatigue, was significantly (P <.01) greater for epoetin alfa versus placebo patients. Although the study was not powered for survival as an end point, Kaplan-Meier estimates showed a trend in overall survival favoring epoetin alfa (P =.13, log-rank test), and Cox regression analysis showed an estimated hazards ratio of 1.309 (P =.052) favoring epoetin alfa. Adverse events were comparable between groups. CONCLUSION: Epoetin alfa safely and effectively ameliorates anemia and significantly improves QOL in cancer patients receiving nonplatinum chemotherapy. Encouraging results regarding increased survival warrant another trial designed to confirm these findings. PMID- 11387361 TI - Patient preferences versus physician perceptions of treatment decisions in cancer care. AB - PURPOSE: To examine patient preferences as well as physician perceptions of these preferences for decision making and communication in palliative care. PATIENTS AND METHODS: Medical decision-making preferences (DMPs) were prospectively studied in 78 assessable cancer patients after initial assessment at a palliative care outpatient clinic. DMPs were assessed with a questionnaire using five possible choices ranging from 1 (patient prefers to make the treatment decision) to 5 (patient prefers the physician to make the decision). In addition, the physician's perception of this preference was assessed. RESULTS: Full concordance between the physician and the patient was seen in 30 (38%) of 78 cases; when the five original categories were recombined to cover active, shared, and passive decision making, there was concordance in 35 (45%) of 78 cases. The kappa coefficient for agreement between physician and patient was poor at 0.14 (95% confidence limit, -0.01 to 0.30) for simple kappa and 0.17 (95% confidence interval [CI], 0.00 to 0.34) for weighted kappa (calculated on the three regrouped categories). Active, shared, and passive DMPs were chosen by 16 (20%) of 78, 49 (63%) of 78, and 13 (17%) of 78 patients, and by 23 (29%) of 78, 30 (39%) of 78, and 25 (32%) of 78 physicians, respectively. The majority of patients (49 [63%] of 78; 95% CI, 0.51 to 0.74) preferred a shared approach with physicians. Physicians predicted that patients preferred a less shared approach than they in fact did. Patient age or sex did not significantly alter DMP. CONCLUSION: An individual approach is needed and each patient should be assessed prospectively for DMP. PMID- 11387360 TI - Clinical evaluation of once-weekly dosing of epoetin alfa in chemotherapy patients: improvements in hemoglobin and quality of life are similar to three times-weekly dosing. AB - PURPOSE: To prospectively evaluate the effectiveness, safety, and clinical benefits of once-weekly epoetin alfa therapy as an adjunct to chemotherapy in anemic cancer patients. PATIENTS AND METHODS: A total of 3,012 patients with nonmyeloid malignancies who received chemotherapy were enrolled onto this multicenter, open-label, nonrandomized study conducted in 600 United States community-based practices. Patients received epoetin alfa 40,000 U once weekly, which could be increased to 60,000 U once weekly after 4 weeks dependent on hemoglobin response. Treatment was continued for a maximum of 16 weeks. RESULTS: Among the 2,964 patients assessable for efficacy, epoetin alfa therapy resulted in significant increases in hemoglobin levels, decreases in transfusion requirements, and improvements in functional status and fatigue as assessed by the linear analog scale assessment (energy level, ability to perform daily activities, and overall quality of life) and the anemia subscale of the Functional Assessment of Cancer Therapy-Anemia questionnaire. Improvements in quality-of-life parameters correlated significantly (P <.001) with increased hemoglobin levels. The direct relationship between hemoglobin and quality-of-life improvement was sustained during the 16-week study period, which is similar to findings of large community-based trials of three-times-weekly dosing. Once weekly epoetin alfa was well tolerated, with most adverse events attributed to the underlying disease or concomitant chemotherapy. CONCLUSION: The results from this large, prospective, community-based trial suggest that once-weekly epoetin alfa therapy increases hemoglobin levels, decreases transfusion requirements, and improves quality of life in patients with cancer and anemia who undergo concomitant chemotherapy. Based on the results of this study, the clinical benefits and the adverse event profile of once-weekly epoetin alfa therapy in community-based practice are similar to those observed in the historical experience with the three-times-weekly dosage schedule. PMID- 11387362 TI - Ensuring quality cancer care by the use of clinical practice guidelines and critical pathways. AB - PURPOSE: We describe the impact of clinical practice guidelines (CPGs) on improvement in oncology treatment processes or outcomes. METHODS: We performed a comprehensive search of the literature from 1966 to the present and a directed review of the literature. RESULTS: Improvements have been demonstrated in compliance with evidence-based guidelines or evidence-based medicine, and in short-term length of stay, complication rates, and financial outcomes. The data suggest that patient satisfaction can be maintained despite a shorter length of stay. There has been one example of province-wide improvement in disease-free and overall survival of breast cancer patients coincident with the adoption of CPGS: The components of successful guidelines can be summarized as follows: (1) development is based on evidence, with the guideline formulated by key physicians in the group; (2) the guidelines are disseminated to all affected health care professionals for critique; (3) implementation includes direct feedback on performance to physicians or general feedback on system performance; and (4) there is accountability for performance according to the guidelines. This accountability can consist of voluntary peer pressure to conform to evidence based medicine, and it does not require a financial reward or penalty. CONCLUSION: Some attempts to improve practice have been moderately successful in achievement of reduced health care costs, reduced hospital length of stay, and possibly improved outcomes. Other methods that are still in use have been demonstrated to have little effect. Programs that have not succeeded have relied on voluntary change in practice behavior without incentives to change or have had no accountability component. Further research is needed to assess how guidelines are enacted in organizations other than those demonstrably committed to improvement, ways to improve compliance of health care providers who are not committed to change, and methods to improve accountability. PMID- 11387363 TI - Switching from morphine to methadone to improve analgesia and tolerability in cancer patients: a prospective study. AB - PURPOSE: To evaluate the clinical benefits of switching from morphine to oral methadone in patients who experience poor analgesia or adverse effects from morphine. PATIENTS AND METHODS: Fifty-two consecutive cancer patients receiving oral morphine but with uncontrolled pain and/or moderate to severe opioid adverse effects were switched to oral methadone administered every 8 hours using different dose ratios. Intensity of pain and adverse effects were assessed daily, and the symptom distress score (DS) was calculated before and after switching. RESULTS: Data were analyzed for 50 patients. Switching was considered effective in 80% of the patients; results were achieved in an average of 3.65 days. In the 10 patients who switched to methadone because of uncontrolled pain, a significant reduction in pain intensity (P <.005) and an average of a 33% increase in methadone doses necessary (P <.01) were found after an average of 3.5 days. DS significantly decreased from an average of 8.4 to 4.5 (P <.0005). In the 32 patients switching because of uncontrolled pain and morphine-related adverse effects, significant improvement was found in pain intensity (P <.0005), nausea and vomiting (P <.03), constipation (P <.001), and drowsiness (P <.01), but a significant increase in the methadone dose of an average of 20% (P <.004) was required. CONCLUSION: In most patients with cancer pain referred for poor pain control and/or adverse effects, switching to oral methadone is a valid therapeutic option. In the clinical setting of poor pain control, higher doses of methadone are necessary with respect to the equianalgesic calculated dose ratios previously published. PMID- 11387364 TI - Low-dose radiation is sufficient for the noninvolved extended-field treatment in favorable early-stage Hodgkin's disease: long-term results of a randomized trial of radiotherapy alone. AB - PURPOSE: To show that radiotherapy (RT) dose to the noninvolved extended field (EF) can be reduced without loss of efficacy in patients with early-stage Hodgkin's disease (HD). PATIENTS AND METHODS: During 1988 to 1994, pathologically staged patients with stage I or II disease who were without risk factors (large mediastinal mass, extranodal lesions, massive splenic disease, elevated erythrocyte sedimentation rate, or three or more involved areas) were recruited from various centers. All patients received 40 Gy total fractionated dose to the involved field areas but were randomly assigned to receive either 40 Gy (arm A) or 30 Gy (arm B) total fractionated dose for the clinically noninvolved EF. No chemotherapy was given. RT films were prospectively reviewed for protocol violations and recurrences retrospectively related to the applied RT. RESULTS: Of 382 recruited patients, 376 were eligible for randomized comparison, 190 in arm A and 186 in arm B. Complete remission was attained in 98% of patients in each arm. With a median follow-up of 86 months, 7-year relapse-free survival (RFS) rates were 78% (arm A) and 83% (arm B) (P =.093). The upper 95% confidence limit for the possible inferiority of arm B in RFS was 4%. Corresponding overall survival rates were 91% (arm A) and 96% (arm B) (P =.16). The most common causes of death (n = 27) were cardiorespiratory disease/pulmonary embolisms (seven), second malignancy (six), and HD (five). Protocol violation was associated with significantly poorer RFS. Nonirradiated nodes were involved in 42 of 52 reviewed relapses, infield areas in 18, marginal areas in 17, and extranodal sites in 16. CONCLUSION: EF-RT alone attains good survival rates in favorable early-stage HD. The 30-Gy dose is adequate for clinically noninvolved areas. Protocol violation worsens the subsequent prognosis. Relapse patterns suggest that systemic therapy can reduce the 20% long-term relapse rate. PMID- 11387365 TI - BCR rearrangement-negative chronic myelogenous leukemia revisited. AB - PURPOSE: To document the characteristics of patients with major breakpoint cluster region (M-bcr) rearrangement-negative chronic myelogenous leukemia (CML). PATIENTS AND METHODS: The hematopathologist, who was blinded to patients' molecular status, reviewed the referral bone marrows and peripheral-blood smears from 26 patients with Philadelphia (Ph) translocation-negative CML who lacked Bcr rearrangement (and other evidence of a Bcr-Abl anomaly) and 14 patients (controls) with chronic-phase Ph-positive CML. Clinical data was ascertained by chart review. RESULTS: Among the 26 M-bcr rearrangement-negative CML patients, three pathologic subtypes emerged: (1) patients indistinguishable from classic CML (n = 9), (2) patients with atypical CML (n = 8), and (3) patients with chronic neutrophilic leukemia (n = 9). Among the 14 patients with Ph-positive CML who were included in the blinded review, 13 were classified as classic CML, and one was classified as atypical CML. The only statistically significant difference between M-bcr rearrangement-negative subgroups was in the proportion of patients having karyotypic abnormalities, an observation common only in patients with atypical CML (P = 0.008). However, the small number of patients in each subgroup limited our ability to differentiate between them. Interferon alfa induced complete hematologic remission in five of 14 patients; four of these remissions lasted more than 5 years. Only one of 26 patients developed blast crisis. The median survival of the 26 patients was 37 months. CONCLUSION: Patients with M-bcr rearrangement-negative CML fall into three morphologic subgroups. Disease evolution does not generally involve blastic transformation. Instead, patients show progressive organomegaly, leukocytosis, anemia, and thrombocytosis. Some patients in each subgroup can respond to interferon alfa. PMID- 11387366 TI - High-dose therapy and autologous stem-cell transplantation versus conventional dose consolidation/maintenance therapy as postremission therapy for adult patients with lymphoblastic lymphoma: results of a randomized trial of the European Group for Blood and Marrow Transplantation and the United Kingdom Lymphoma Group. AB - PURPOSE: To determine whether a combination of high-dose therapy and autologous stem-cell transplantation (ASCT) is superior to conventional-dose consolidation and maintenance chemotherapy as postremission therapy in adults with lymphoblastic lymphoma. PATIENTS AND METHODS: One hundred nineteen patients were entered onto this prospective randomized trial from 37 centers. Patients received standard remission induction therapy, and responding patients were randomized either to continue with a conventional consolidation/maintenance protocol (CC) or to receive high-dose therapy and ASCT. In some centers, patients with HLA identical sibling donors were registered on the trial but proceeded to allogeneic bone marrow transplantation (BMT) without randomization. RESULTS: Of the 119 patients entered, 111 were assessable for response to induction therapy. The overall response rate was 82% (56% complete response, 26% partial response). Of the 98 patients eligible for randomization, 65 were randomized, 31 to ASCT and 34 to CC. Reasons for failure to randomize included patient refusal (12 patients), early progression or death on induction therapy (eight patients), excessive toxicity of induction regimen (six patients), and elective allogeneic BMT (12 patients). With a median follow-up of 37 months, the actuarial 3-year relapse free survival rate is 24% for the CC arm and 55% for the ASCT arm (hazards ratio = 0.55 in favor of the ASCT arm; 95% confidence interval [CI], 0.29 to 1.04; P =.065). The corresponding figures for overall survival are 45% and 56%, respectively (hazards ratio = 0.87 in favor of the ASCT arm; 95% CI, 0.42 to 1.81; P =.71). CONCLUSION: The use of ASCT in adults with lymphoblastic lymphoma in first remission produced a trend for improved relapse-free survival but did not improve overall survival compared with conventional-dose therapy in this small randomized trial. PMID- 11387368 TI - Gene discovery using the serial analysis of gene expression technique: implications for cancer research. AB - Cancer is a genetic disease. As such, our understanding of the pathobiology of tumors derives from analyses of the genes whose mutations are responsible for those tumors. The cancer phenotype, however, likely reflects the changes in the expression patterns of hundreds or even thousands of genes that occur as a consequence of the primary mutation of an oncogene or a tumor suppressor gene. Recently developed functional genomic approaches, such as DNA microarrays and serial analysis of gene expression (SAGE), have enabled researchers to determine the expression level of every gene in a given cell population, which represents that cell population's entire transcriptome. The most attractive feature of SAGE is its ability to evaluate the expression pattern of thousands of genes in a quantitative manner without prior sequence information. This feature has been exploited in three extremely powerful applications of the technology: the definition of transcriptomes, the analysis of differences between the gene expression patterns of cancer cells and their normal counterparts, and the identification of downstream targets of oncogenes and tumor suppressor genes. Comprehensive analyses of gene expression not only will further understanding of growth regulatory pathways and the processes of tumorigenesis but also may identify new diagnostic and prognostic markers as well as potential targets for therapeutic intervention. PMID- 11387367 TI - Phase I and pharmacokinetic study of NSC 655649, a rebeccamycin analog with topoisomerase inhibitory properties. AB - PURPOSE: To assess the feasibility of administering NSC 655649, a water-soluble, rebeccamycin analog with topoisomerase inhibitory properties, as a brief intravenous (IV) infusion once every 3 weeks and to determine the maximum tolerated dose (MTD) of NSC 655649, characterize its pharmacokinetic behavior, and seek preliminary evidence of antitumor activity. PATIENTS AND METHODS: Patients with advanced solid malignancies were treated with escalating doses of NSC 655649 administered over 30 to 60 minutes IV once every 3 weeks. An accelerated dose-escalation method was used to guide dose escalation. After three patients were treated at the first dose level, doses were escalated in increments that ranged up to 150% using single patient cohorts until moderate toxicity was observed, when a more conservative dose-escalation scheme was invoked. MTD was defined as the highest dose level at which the incidence of dose-limiting toxicity did not exceed 20%. MTD was determined for both minimally pretreated (MP) and heavily pretreated (HP) patients. Plasma and urine were sampled to characterize the pharmacokinetic and excretory behavior of NSC 655649. RESULTS: Forty-five patients were treated with 130 courses of NSC 655649 at doses ranging from 20 mg/m(2) to 744 mg/m(2). Myelosuppression was the principal toxicity. Severe neutropenia, which was often associated with thrombocytopenia, was unacceptably high in HP and MP patients treated at 572 mg/m(2) and 744 mg/m(2), respectively. Nausea, vomiting, and diarrhea were common but rarely severe. The pharmacokinetics of NSC 655649 were dose dependent and fit a three-compartment model. The clearance and terminal elimination half-lives for NSC 655649 averaged 7.57 (SD = 4.2) L/h/m(2) and 48.85 (SD = 23.65) hours, respectively. Despite a heterogeneous population of MP and HP patients, the magnitude of drug exposure correlated well with the severity of myelosuppression. Antitumor activity was observed in two HP ovarian cancer patients and one patient with a soft tissue sarcoma refractory to etoposide and doxorubicin. CONCLUSION: Recommended phase II doses are 500 mg/m(2) and 572 mg/m(2) IV once every 3 weeks for HP and MP patients, respectively. The absence of severe nonhematologic toxicities, the encouraging antitumor activity in HP patients, and the unique mechanism of antineoplastic activity of NSC 655649 warrant further clinical development. PMID- 11387369 TI - Unusual locations for lymphomas. Case 1. Intermediate-grade lymphoma of the cervix. PMID- 11387370 TI - Unusual locations for lymphomas. Case 2. Pineal lymphoma. PMID- 11387371 TI - Unusual locations for lymphomas. Case 3. Successive occurrence of peripheral T cell lymphoma with bilateral conjuctival involvement in a patient with low-grade B-cell lymphoma. PMID- 11387372 TI - First, you cry, 25 years later. PMID- 11387373 TI - Bone mass after treatment for acute lymphoblastic leukemia in childhood. PMID- 11387374 TI - Chronic expanding hematoma mimicking soft tissue neoplasm. PMID- 11387377 TI - Retraction. PMID- 11387375 TI - Correct fluorouracil (5-FU) half-life comparator for a 5-FU prodrug plus a dihydropyrimidine dehydrogenase inhibitor. PMID- 11387378 TI - Auditory edge detection: a neural model for physiological and psychoacoustical responses to amplitude transients. AB - Primary segmentation of visual scenes is based on spatiotemporal edges that are presumably detected by neurons throughout the visual system. In contrast, the way in which the auditory system decomposes complex auditory scenes is substantially less clear. There is diverse physiological and psychophysical evidence for the sensitivity of the auditory system to amplitude transients, which can be considered as a partial analogue to visual spatiotemporal edges. However, there is currently no theoretical framework in which these phenomena can be associated or related to the perceptual task of auditory source segregation. We propose a neural model for an auditory temporal edge detector, whose underlying principles are similar to classical visual edge detector models. Our main result is that this model reproduces published physiological responses to amplitude transients collected at multiple levels of the auditory pathways using a variety of experimental procedures. Moreover, the model successfully predicts physiological responses to a new set of amplitude transients, collected in cat primary auditory cortex and medial geniculate body. Additionally, the model reproduces several published psychoacoustical responses to amplitude transients as well as the psychoacoustical data for amplitude edge detection reported here for the first time. These results support the hypothesis that the response of auditory neurons to amplitude transients is the correlate of psychoacoustical edge detection. PMID- 11387379 TI - Properties of synchronous and asynchronous release during pulse train depression in cultured hippocampal neurons. AB - Neurotransmitter release displays at least two kinetically distinct components in response to a single action potential. The majority of release occurs synchronously with action-potential-triggered Ca(2+) influx; however, delayed release--also called asynchronous release--persists for tens of milliseconds following the peak Ca(2+) transient. In response to trains of action potentials, synchronous release eventually declines, whereas asynchronous release often progressively increases, an effect that is primarily attributed to the buildup of intracellular Ca(2+) during repetitive stimulation. The precise relationship between synchronous and asynchronous release remains unclear at central synapses. To gain better insight into the mechanisms that regulate neurotransmitter release, we systematically characterized the two components of release during repetitive stimulation at excitatory autaptic hippocampal synapses formed in culture. Manipulations that increase the Ca(2+) influx triggered by an action potential--elevation of extracellular Ca(2+) or bath application of tetraethylammonium (TEA)--accelerated the progressive decrease in synchronous release (peak excitatory postsynaptic current amplitude) and concomitantly increased asynchronous release. When intracellular Ca(2+) was buffered by extracellular application of EGTA-AM, initial depression of synchronous release was equal to or greater than control; however, it quickly reached a plateau without further depression. In contrast, asynchronous release was largely abolished in EGTA-AM. The total charge transfer following each pulse--accounting for both synchronous and asynchronous release--reached a steady-state level that was similar between control and EGTA-AM. A portion of the decreased synchronous release in control conditions therefore was matched by a higher level of asynchronous release. We also examined the relative changes in synchronous and asynchronous release during repetitive stimulation under conditions that highly favor asynchronous release by substituting extracellular Ca(2+) with Sr(2+). Initially, asynchronous release was twofold greater in Sr(2+). By the end of the train, the difference was approximately 50%; consequently, the total release per pulse during the plateau phase was slightly larger in Sr(2+) compared with Ca(2+). We thus conclude that while asynchronous release--like synchronous release--is limited by vesicle availability, it may be able to access a slightly larger subset of the readily releasable pool. Our results are consistent with the view that during repetitive stimulation, the elevation of asynchronous release depletes the vesicles immediately available for release, resulting in depression of synchronous release. This implies that both forms of release share a small pool of immediately releasable vesicles, which is being constantly depleted and refilled during repetitive stimulation. PMID- 11387380 TI - Electrophysiological characteristics of rat gustatory cyclic nucleotide--gated channel expressed in Xenopus oocytes. AB - The complementary DNA encoding gustatory cyclic nucleotide--gated ion channel (or gustCNG channel) cloned from rat tongue epithelial tissue was expressed in Xenopus oocytes, and its electrophysiological characteristics were investigated using tight-seal patch-clamp recordings of single and macroscopic channel currents. Both cGMP and cAMP directly activated gustCNG channels but with markedly different affinities. No desensitization or inactivation of gustCNG channel currents was observed even in the prolonged application of the cyclic nucleotides. Single-channel conductance of gustCNG channel was estimated as 28 pS in 130 mM of symmetric Na(+). Single-channel current recordings revealed fast open-close transitions and longer lasting closure states. The distribution of both open and closed events could be well fitted with two exponential components and intracellular cGMP increased the open probability (P(o)) of gustCNG channels mainly by increasing the slower opening rate. Under bi-ionic conditions, the selectivity order of gustCNG channel among divalent cations was determined as Na(+) approximately K(+) > Rb(+) > Li(+) > Cs(+) with the permeability ratio of 1:0.95:0.74:0.63:0.49. Magnesium ion blocked Na(+) currents through gustCNG channels from both intracellular and extracellular sides in voltage-dependent manners. The inhibition constants (K(i)s) of intracellular Mg(2+) were determined as 360 +/- 40 microM at 70 mV and 8.2 +/- 1.5 mM at -70 mV with z delta value of 1.04, while K(i)s of extracellular Mg(2+) were as 1.1 +/- 0.3 mM at 70 mV and 20.0 +/- 0.1 microM at -70 mV with z delta of 0.94. Although 100 microM l-cis diltiazem blocked significant portions of outward Na(+) currents through both bovine rod and rat olfactory CNG channels, the gustCNG channel currents were minimally affected by the same concentration of the drug. PMID- 11387381 TI - Role of mammalian auditory cortex in the perception of elementary sound properties. AB - Studies in several mammalian species have demonstrated that bilateral ablations of the auditory cortex have little effect on simple sound intensity and frequency based behaviors. In the rat, for example, early experiments have shown that auditory ablations result in virtually no effect on the rat's ability to either detect tones or discriminate frequencies. Such lesion experiments, however, typically examine an animal's performance some time after recovery from ablation surgery. As such, they demonstrate that the cortex is not essential for simple auditory behaviors in the long run. Our study further explores the role of cortex in basic auditory perception by examining whether the cortex is normally involved in these behaviors. In these experiments we reversibly inactivated the rat primary auditory cortex (AI) using the GABA agonist muscimol, while the animals performed a simple auditory task. At the same time we monitored the rat's auditory activity by recording auditory evoked potentials (AEP) from the cortical surface. In contrast to lesion studies, the rapid time course of these experimental conditions preclude reorganization of the auditory system that might otherwise compensate for the loss of cortical processing. Soon after bilateral muscimol application to their AI region, our rats exhibited an acute and profound inability to detect tones. After a few hours this state was followed by a gradual recovery of normal hearing, first of tone detection and, much later, of the ability to discriminate frequencies. Surface muscimol application, at the same time, drastically altered the normal rat AEP. Some of the normal AEP components vanished nearly instantaneously to unveil an underlying waveform, whose size was related to the severity of accompanying behavioral deficits. These results strongly suggest that the cortex is directly involved in basic acoustic processing. Along with observations from accompanying multiunit experiments that related the AEP to AI neuronal activity, our results suggest that a critical amount of activity in the auditory cortex is necessary for normal hearing. It is likely that the involvement of the cortex in simple auditory perceptions has hitherto not been clearly understood because of underlying recovery processes that, in the long-term, safeguard fundamental auditory abilities after cortical injury. PMID- 11387382 TI - Cerebellar cortical stimulation increases spinal visceral nociceptive responses. AB - The role of the cerebellum in modulating nociceptive phenomena is unclear. In this study, we focus on the effects of cerebellar cortical stimulation on the responses of midline neurons of the lumbosacral spinal cord to graded nonnoxious and noxious visceral (colorectal distension) as well as somatic (brush, pressure, pinch) stimuli. Extracellular recording was used for the isolation and recording of spinal nociceptive neurons, while electrical current pulses and chemical injection of D, L-homocysteic acid were used to stimulate the cortex of the posterior cerebellar vermis. Cerebellar cortical stimulation increased the responses of all isolated cells to colorectal distension, whereas the effect on the responses to somatic stimuli was variable. These findings indicate that the posterior cerebellar vermis may exert a pro-nociceptive effect on spinal visceroceptive neurons. PMID- 11387383 TI - Neural representations of temporally asymmetric stimuli in the auditory cortex of awake primates. AB - The representation of rapid acoustic transients by the auditory cortex is a fundamental issue that is still unresolved. Auditory cortical neurons have been shown to be limited in their stimulus-synchronized responses, yet the perceptual performances of humans and animals in discriminating temporal variations in complex sounds are better than what existing neurophysiological data would predict. This study investigated the neural representation of temporally asymmetric stimuli in the primary auditory cortex of awake marmoset monkeys. The stimuli, ramped and damped sinusoids, were systematically manipulated (by means of half-life of the exponential envelope) within a cortical neuron's presumed temporal integration window. The main findings of this study are as follows: 1) temporal asymmetry in ramped and damped sinusoids with a short period (25 ms) was clearly reflected by average discharge rate but not necessarily by temporal discharge patterns of auditory cortical neurons. There was considerable response specificity to these stimuli such that some neurons were strongly responsive to a ramped sinusoid but almost completely unresponsive to its damped counterpart or vice versa. Of 181 neurons studied, 140 (77%) showed significant response asymmetry in at least one of the tested half-life values of the exponential envelope. Forty-six neurons showed significant response asymmetry over all half lives tested. Sustained firing, commonly observed under awake conditions, contributed to greater response asymmetry than that of onset responses in many neurons. 2) A greater proportion of the neurons (32/46) that exhibited significant overall response asymmetry showed stronger responses to the ramped sinusoids than to the damped sinusoids, possibly contributing to the difference in the perceived loudness between these two classes of sounds. 3) The asymmetry preference of a neuron to ramped or damped sinusoids did not appear to be correlated with its characteristic frequency or minimum response latency, suggesting that this is a general phenomenon that exists across populations of cortical neurons. Moreover, the intensity of the stimuli did not have significant effects on the measure of the asymmetry preference based on discharge rate. 4) A population measure of response preference, based on discharge rate, of cortical neurons to the temporally asymmetric stimuli was qualitatively similar to the performance of human listeners in discriminating ramped versus damped sinusoids at different half-life values. These findings suggest that rapid acoustic transients embedded in complex sounds can be represented by discharge rates of cortical neurons instead of or in the absence of stimulus-synchronized discharges. PMID- 11387384 TI - GABA(A)-dependent chloride influx modulates reversal potential of GABA(B) mediated IPSPs in hippocampal pyramidal cells. AB - Changes in intracellular chloride concentration, mediated by chloride influx through GABA(A) receptor-gated channels, may modulate GABA(B) receptor-mediated inhibitory postsynaptic potentials (GABA(B) IPSPs) via unknown mechanisms. Recording from CA3 pyramidal cells in hippocampal slices, we investigated the impact of chloride influx during GABA(A) receptor-mediated IPSPs (GABA(A) IPSPs) on the properties of GABA(B) IPSPs. At relatively positive membrane potentials (near -55 mV), mossy fiber--evoked GABA(B) IPSPs were reduced (compared with their magnitude at -60 mV) when preceded by GABA(A) receptor--mediated chloride influx. This effect was not associated with a correlated reduction in membrane permeability during the GABA(B) IPSP. The mossy fiber--evoked GABA(B) IPSP showed a positive shift in reversal potential (from -99 to -93 mV) when it was preceded by a GABA(A) IPSP evoked at cell membrane potential of -55 mV as compared with 60 mV. Similarly, when intracellular chloride concentration was raised via chloride diffusion from an intracellular microelectrode, there was a reduction of the pharmacologically isolated monosynaptic GABA(B) IPSP and a concurrent shift of GABA(B) IPSP reversal potential from -98 to -90 mV. We conclude that in hippocampal pyramidal cells, in which "resting" membrane potential is near action potential threshold, chloride influx via GABA(A) IPSPs shifts the reversal potential of subsequent GABA(B) receptor--mediated postsynaptic responses in a positive direction and reduces their magnitude. PMID- 11387385 TI - Local opiate withdrawal in locus coeruleus neurons in vitro. AB - Noradrenergic neurons of the brain nucleus locus coeruleus (LC) become hyperactive during opiate withdrawal. It has been uncertain to what extent such hyperactivity reflects changes in intrinsic properties of these cells. The effects of withdrawal from chronic morphine on the activity of LC neurons were studied using intracellular recordings in rat brain slices. LC neurons in slices from chronically morphine-treated rats exhibited more than twice the frequency of spontaneous action potentials after naloxone compared with LC neurons from control rats. However, after naloxone treatment, the resting membrane potential (MP) of LC neurons from dependent rats was not significantly different from that in control rats. Neither resting MP nor spontaneous discharge rate (SDR) was altered by naloxone in LC neurons from control rats. Neither kynurenic acid nor a cocktail of glutamate and GABA antagonists (6-cyano-7-nitroquinoxalene-2,3-dione + 2-amino-5-phosphonopentanoic acid + bicuculline) blocked the hyperactivity of LC neurons precipitated by naloxone in slices from morphine-dependent rats. The effects of ouabain on MP and SDR were similar in LC neurons from control and morphine-dependent rats. These results indicate that an adaptive change in glutamatergic or GABAergic synaptic mechanisms or altered Na/K pump activity does not underlie the withdrawal-induced activation of LC neurons in vitro. Specific inhibitors of protein kinase A [Rp-cAMPS or N-(2-[p-bromocinnamylamino]ethyl)-5 isoquinolinesulfonamide (H-89)] partially suppressed the withdrawal hyperactivity of LC neurons, and activators of cAMP (forskolin) or protein kinase A (Sp-cAMPS) increased the discharge rate of LC neurons from control rats. These results suggest that upregulation of cAMP-dependent protein kinase A during chronic morphine treatment is involved in the withdrawal-induced hyperactivity of LC neurons. PMID- 11387386 TI - Modulation of excitability in Aplysia tail sensory neurons by tyrosine kinases. AB - Tyrosine kinases have recently been shown to modulate synaptic plasticity and ion channel function. We show here that tyrosine kinases can also modulate both the baseline excitability state of Aplysia tail sensory neurons (SNs) as well as the excitability induced by the neuromodulator serotonin (5HT). First, we examined the effects of increasing and decreasing tyrosine kinase activity in the SNs. We found that tyrosine kinase inhibitors decrease baseline SN excitability in addition to attenuating the increase in excitability induced by 5HT. Conversely, functionally increasing cellular tyrosine kinase activity in the SNs by either inhibiting opposing tyrosine phosphatase activity or by direct injection of an active tyrosine kinase (Src) induces increases in SN excitability in the absence of 5HT. Second, we examined the interaction between protein kinase A (PKA), which is known to mediate 5HT-induced excitability changes in the SNs, and tyrosine kinases, in the enhancement of SN excitability. We found that the tyrosine kinases function downstream of PKA activation since tyrosine kinase inhibitors reduce excitability induced by activators of PKA. Finally, we examined the role of tyrosine kinases in other forms of 5HT-induced plasticity in the SNs. We found that while tyrosine kinase inhibitors attenuate excitability produced by 5HT, they have no effect on short-term facilitation (STF) of the SN-motor neuron (MN) synapse induced by 5HT. Thus tyrosine kinases modulate different forms of SN plasticity independently. Such differential modulation would have important consequences for activity-dependent plasticity in a variety of neural circuits. PMID- 11387387 TI - Dynamic modulation of excitation and inhibition during stimulation at gamma and beta frequencies in the CA1 hippocampal region. AB - Fast oscillations at gamma and beta frequency are relevant to cognition. During this activity, excitatory and inhibitory postsynaptic potentials (EPSPs and IPSPs) are generated rhythmically and synchronously and are thought to play an essential role in pacing the oscillations. The dynamic changes occurring to excitatory and inhibitory synaptic events during repetitive activation of synapses are therefore relevant to fast oscillations. To cast light on this issue in the CA1 region of the hippocampal slice, we used a train of stimuli, to the pyramidal layer, comprising 1 s at 40 Hz followed by 2--3 s at 10 Hz, to mimic the frequency pattern observed during fast oscillations. Whole cell current-clamp recordings from CA1 pyramidal neurons revealed that individual stimuli at 40 Hz produced EPSPs riding on a slow biphasic hyperpolarizing-depolarizing waveform. EPSP amplitude initially increased; it then decreased concomitantly with the slow depolarization and with a large reduction in membrane resistance. During the subsequent 10-Hz train: the cells repolarized, EPSP amplitude and duration increased to above control, and no IPSPs were detected. In the presence of GABA(A) receptor antagonists, the slow depolarization was blocked, and EPSPs of constant amplitude were generated by 10-Hz stimuli. Altering pyramidal cell membrane potential affected the time course of the slow depolarization, with the peak being reached earlier at more negative potentials. Glial recordings revealed that the trains were associated with extracellular potassium accumulation, but the time course of this event was slower than the neuronal depolarization. Numerical simulations showed that intracellular chloride accumulation (due to massive GABAergic activation) can account for these observations. We conclude that synchronous activation of inhibitory synapses at gamma frequency causes a rapid chloride accumulation in pyramidal neurons, decreasing the efficacy of inhibitory potentials. The resulting transient disinhibition of the local network leads to a short-lasting facilitation of polysynaptic EPSPs. These results set constraints on the role that synchronous, rhythmic IPSPs may play in pacing oscillations at gamma frequency in the CA1 hippocampal region. PMID- 11387388 TI - Effects of adult neurogenesis on synaptic plasticity in the rat dentate gyrus. AB - Ongoing neurogenesis in the adult hippocampal dentate gyrus (DG) generates a substantial population of young neurons. This phenomenon is present in all species examined thus far, including humans. Although the regulation of adult neurogenesis by various physiologically relevant factors such as learning and stress has been documented, the functional contributions of the newly born neurons to hippocampal functions are not known. We investigated possible contributions of the newly born granule neurons to synaptic plasticity in the hippocampal DG. In the standard hippocampal slice preparation perfused with artificial cerebrospinal fluid (ACSF), a small (10%) long-term potentiation (LTP) of the evoked field potentials is seen after tetanic stimulation of the afferent medial perforant pathway (MPP). The induction of this ACSF-LTP is resistant to a N-methyl-D-aspartate (NMDA) receptor blocker, D,L-2-amino-5-phosphonovaleric acid (APV), but is completely prevented by ifenprodil, a blocker of NR2B subtype of NMDA receptors. In contrast, slices perfused with picrotoxin (PICRO), a GABA receptor blocker, revealed a larger (40--50%), APV-sensitive but ifenprodil insensitive LTP. The ACSF-LTP required lower frequency of stimulation and fewer stimuli for its induction than the PICRO-LTP. All these characteristics of ACSF LTP are in agreement with the properties of the putative individual new granule neurons examined previously with the use of the whole cell recording technique in a similar preparation. A causal relationship between neurogenesis and ACSF-LTP was confirmed in experiments using low dose of gamma radiation applied to the brain 3 wk prior to the electrophysiological experiments. In these experiments, the new cell proliferation was drastically reduced and ACSF-LTP was selectively blocked. We conclude that the young, adult-generated granule neurons play a significant role in synaptic plasticity in the DG. Since DG is the major source of the afferent inputs into the hippocampus, the production and the plasticity of new neurons may have an important role in the hippocampal functions such as learning and memory. PMID- 11387389 TI - Neural mechanisms for generating rate and temporal codes in model CA3 pyramidal cells. AB - The effect of synaptic inhibition on burst firing of a two-compartment model of a CA3 pyramidal cell is considered. We show that, depending on its timing, a short dose of fast decaying synaptic inhibition can either delay or advance the timing of firing of subsequent bursts. Moreover, increasing the strength of the inhibitory input is shown to modulate the burst profile from a full complex burst, to a burst with multiple spikes, to single spikes. We additionally show how slowly decaying inhibitory input can be used to synchronize a network of pyramidal cells. Implications for the phase precession phenomenon of hippocampal place cells and for the generation of temporal and rate codes are discussed. PMID- 11387390 TI - Characteristics of glutamate-evoked temporomandibular joint afferent activity in the rat. AB - Injection of glutamate into the rat temporomandibular joint (TMJ) capsule can reflexly induce a prolonged increase in the electromyographic (EMG) activity of the jaw muscles, however, the characteristics of TMJ afferents activated by glutamate have not been investigated. In the present study, we examined the effect of glutamate injection into the TMJ capsule on jaw muscle EMG activity and the extracellularly recorded activity of single trigeminal afferents that had receptive fields in the TMJ tissue and antidromically identified projections to the brain stem subnucleus caudalis (Vc) in rats of both sexes. Glutamate (0.05- 1.0 M, 10 microl) injection into the TMJ capsule evoked EMG activity in a dose related manner; however, at concentrations of 0.5 and 1.0 M, glutamate-evoked digastric muscle responses were greater in female than in male rats. In experiments where jaw muscle EMG and afferent activity were recorded simultaneously, glutamate (0.5 M, 10 microl) injection into the TMJ capsule evoked activity in the jaw muscles as well as in 27 (26 A delta and 1 C-fiber afferent) of 34 trigeminal afferents that could be activated by blunt mechanical stimulation of the TMJ tissue. In these experiments, glutamate-evoked jaw muscle activity was significantly increased for 6 min after the glutamate injection, whereas afferent activity was significantly increased only during the first minute after the glutamate injection. The glutamate-evoked afferent activity was inversely related to conduction velocity and, in afferents with conduction velocities <10 m/s, was significantly greater in female (n = 6) than in male (n = 10) rats. These results suggest that glutamate excites putative nociceptive afferents within the TMJ to a greater degree in female than in male rats. This sex-related difference in afferent discharge may, in part, underlie sex-related differences in glutamate-evoked jaw muscle EMG activity. PMID- 11387391 TI - Gravitoinertial force magnitude and direction influence head-centric auditory localization. AB - We measured the influence of gravitoinertial force (GIF) magnitude and direction on head-centric auditory localization to determine whether a true audiogravic illusion exists. In experiment 1, supine subjects adjusted computer-generated dichotic stimuli until they heard a fused sound straight ahead in the midsagittal plane of the head under a variety of GIF conditions generated in a slow-rotation room. The dichotic stimuli were constructed by convolving broadband noise with head-related transfer function pairs that model the acoustic filtering at the listener's ears. These stimuli give rise to the perception of externally localized sounds. When the GIF was increased from 1 to 2 g and rotated 60 degrees rightward relative to the head and body, subjects on average set an acoustic stimulus 7.3 degrees right of their head's median plane to hear it as straight ahead. When the GIF was doubled and rotated 60 degrees leftward, subjects set the sound 6.8 degrees leftward of baseline values to hear it as centered. In experiment 2, increasing the GIF in the median plane of the supine body to 2 g did not influence auditory localization. In experiment 3, tilts up to 75 degrees of the supine body relative to the normal 1 g GIF led to small shifts, 1--2 degrees, of auditory setting toward the up ear to maintain a head-centered sound localization. These results show that head-centric auditory localization is affected by azimuthal rotation and increase in magnitude of the GIF and demonstrate that an audiogravic illusion exists. Sound localization is shifted in the direction opposite GIF rotation by an amount related to the magnitude of the GIF and its angular deviation relative to the median plane. PMID- 11387393 TI - H(2)O(2) is a novel, endogenous modulator of synaptic dopamine release. AB - Recent evidence suggests that reactive oxygen species (ROS) might act as modulators of neuronal processes, including synaptic transmission. Here we report that synaptic dopamine (DA) release can be modulated by an endogenous ROS, H(2)O(2). Electrically stimulated DA release was monitored in guinea pig striatal slices using carbon-fiber microelectrodes with fast-scan cyclic voltammetry. Exogenously applied H(2)O(2) reversibly inhibited evoked release in the presence of 1.5 mM Ca(2+). The effectiveness of exogenous H(2)O(2), however, was abolished or decreased by conditions that enhance Ca(2+) entry, including increased extracellular Ca(2+) concentration ([Ca(2+)](o); to 2.4 mM), brief, high frequency stimulation, and blockade of inhibitory D(2) autoreceptors. To test whether DA release could be modulated by endogenous H(2)O(2), release was evoked in the presence of the H(2)O(2)-scavenging enzyme, catalase. In the presence of catalase, evoked [DA](o) was 60% higher than after catalase washout, demonstrating that endogenously generated H(2)O(2) can also inhibit DA release. Importantly, the Ca(2+) dependence of the catalase-mediated effect was opposite to that of H(2)O(2): catalase had a greater enhancing effect in 2.4 mM Ca(2+) than in 1.5 mM, consistent with enhanced H(2)O(2) generation in higher [Ca(2+)](o). Together these data suggest that H(2)O(2) production is Ca(2+) dependent and that the inhibitory mechanism can be saturated, thus preventing further effects from exogenous H(2)O(2). These findings show for the first time that endogenous H(2)O(2) can modulate vesicular neurotransmitter release, thus revealing an important new signaling role for ROS in synaptic transmission. PMID- 11387392 TI - Mechanisms underlying regulation of respiratory pattern by nicotine in preBotzinger complex. AB - Cholinergic neurotransmission plays a role in regulation of respiratory pattern. Nicotine from cigarette smoke affects respiration and is a risk factor for sudden infant death syndrome (SIDS) and sleep-disordered breathing. The cellular and synaptic mechanisms underlying this regulation are not understood. Using a medullary slice preparation from neonatal rat that contains the preBotzinger Complex (preBotC), the hypothesized site for respiratory rhythm generation, and generates respiratory-related rhythm in vitro, we examined the effects of nicotine on excitatory neurotransmission affecting inspiratory neurons in preBotC and on the respiratory-related motor activity from hypoglossal nerve (XIIn). Microinjection of nicotine into preBotC increased respiratory frequency and decreased the amplitude of inspiratory bursts, whereas when injected into XII nucleus induced a tonic activity and an increase in amplitude but not in frequency of inspiratory bursts from XIIn. Bath application of nicotine (0.2--0.5 microM, approximately the arterial blood nicotine concentration immediately after smoking a cigarette) increased respiratory frequency up to 280% of control in a concentration-dependent manner. Nicotine decreased the amplitude to 82% and increased the duration to 124% of XIIn inspiratory bursts. In voltage-clamped preBotC inspiratory neurons (including neurons with pacemaker properties), nicotine induced a tonic inward current of -19.4 +/- 13.4 pA associated with an increase in baseline noise. Spontaneous excitatory postsynaptic currents (sEPSCs) present during the expiratory period increased in frequency to 176% and in amplitude to 117% of control values; the phasic inspiratory drive inward currents decreased in amplitude to 66% and in duration to 89% of control values. The effects of nicotine were blocked by mecamylamine (Meca). The inspiratory drive current and sEPSCs were completely eliminated by 6-cyano-7-nitroquinoxaline-2,3 dione (CNQX) in the presence or absence of nicotine. In the presence of tetrodotoxin (TTX), low concentrations of nicotine did not induce any tonic current or any increase in baseline noise, nor affect the input resistance in inspiratory neurons. In this study, we demonstrated that nicotine increased respiratory frequency and regulated respiratory pattern by modulating the excitatory neurotransmission in preBotC. Activation of nicotinic acetylcholine receptors (nAChRs) enhanced the tonic excitatory synaptic input to inspiratory neurons including pacemaker neurons and at the same time, inhibited the phasic excitatory coupling between these neurons. These mechanisms may account for the cholinergic regulation of respiratory frequency and pattern. PMID- 11387394 TI - Influence of expectation of different rewards on behavior-related neuronal activity in the striatum. AB - This study investigated how different expected rewards influence behavior-related neuronal activity in the anterior striatum. In a spatial delayed-response task, monkeys reached for a left or right target and obtained a small quantity of one of two juices (apple, grenadine, orange, lemon, black currant, or raspberry). In each trial, an initial instruction picture indicated the behavioral target and predicted the reward. Nonmovement trials served as controls for movement relationships. Consistent preferences in special reward choice trials and differences in anticipatory licks, performance errors, and reaction times indicated that animals differentially expected the rewards predicted by the instructions. About 600 of >2,500 neurons in anterior parts of caudate nucleus, putamen, and ventral striatum showed five forms of task-related activations, comprising responses to instructions, spatial or nonspatial activations during the preparation or execution of the movement, and activations preceding or following the rewards. About one-third of the neurons showed different levels of task-related activity depending on which liquid reward was predicted at trial end. Activations were either higher or lower for rewards that were preferred by the animals as compared with nonpreferred rewards. These data suggest that the expectation of an upcoming liquid reward may influence a fraction of task-related neurons in the anterior striatum. Apparently the information about the expected reward is incorporated into the neuronal activity related to the behavioral reaction leading to the reward. The results of this study are in general agreement with an account of goal-directed behavior according to which the outcome should be represented already at the time at which the behavior toward the outcome is performed. PMID- 11387395 TI - Group I mGluRs coupled to G proteins are regulated by tyrosine kinase in dopamine neurons of the rat midbrain. AB - Metabotropic glutamate receptors (mGluRs) modulate neuronal function via different transduction mechanisms that are either dependent or independent on G protein function. Here we investigated, using whole cell patch-clamp recordings in combination with fluorimetric measurements of intracellular calcium concentration ([Ca(2+)](i)), the metabolic pathways involved in the responses induced by group I mGluRs in dopamine neurons of the rat midbrain. The inward current and the [Ca(2+)](i) increase caused by the group I mGluR agonist (S)-3,5 dihydroxyphenylglycine (DHPG, 100 microM) were permanently activated and subsequently abolished in cells loaded with the nonhydrolizable GTP-analogue GTP gamma-S (600 microM). In addition, when GDP-beta-S (600 microM) was dialyzed into the cells to produce the blockade of the G proteins, the DHPG-dependent responses were reduced. When the tissue was bathed with the phospholipase C inhibitor 1 [6[[(17 beta)-3-methoxyestra-1,3,5(10)-trien-17-yl]amino]exyl]-1H-pyrrole-2,5 dione (10 microM), the DHPG-induced calcium transients slightly diminished but the associated inward currents were not affected. Interestingly, a substantial depression of the DHPG-induced inward current and transient increase of [Ca(2+)](i) was caused by the protein tyrosine kinase inhibitors tyrphostin B52 (40 microM) and 4',5,7-trihydroxyisoflavone (genistein; 40 microM), whereas genistein's inactive analogue 4',5,7-trihydroxyisoflavone-7-glucoside (40 microM) was ineffective. The blockade of the Src family of tyrosine kinase by 4-amino-5 (4-methylphenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine (20 microM), mitogen activated protein kinase by 2'-amino-3' methoxyflavone (50 microM), and protein kinase C by staurosporine (1 microM) had no effect on the cellular responses caused by DHPG. The mGluR5-selective antagonist 2-methyl-6-(phenylethynyl) pyridine (10--100 microM) did not affect the actions of DHPG. Thus our results indicate that the responses, mainly mediated by mGluRs1 in dopamine neurons, are activated by intracellular mechanisms coupled to G proteins and regulated by tyrosine kinases. PMID- 11387396 TI - Differential modulation of nicotinic acetylcholine receptor subtypes and synaptic transmission in chick sympathetic ganglia by PGE(2). AB - The diversity of neuronal nicotinic acetylcholine receptors (nAChRs) is likely an important factor in the modulation of synaptic transmission by acetylcholine and nicotine. We have tested whether postsynaptic nAChRs are modulated in a subtype specific manner by prostaglandin E(2) (PGE(2)), a regulator of neuronal excitability in both the central and peripheral nervous systems, and examined the effects of PGE(2) on nicotinic transmission. Somatodendritic nAChRs in chick lumbar sympathetic ganglia include four nAChR subtypes distinguished on the basis of conductance and kinetic profile. Nanomolar PGE(2) applied to the extrapatch membrane differentially regulates opening probability (Po), frequency and the opening duration of each nAChR channel subtype in cell-attached patches. PGE(2) decreases the Po of the predominant nAChR subtype (36 pS) and significantly increases Po and open duration of the 23 pS subtype. The 23 pS subtype is gated by the alpha 7-selective agonist choline, and choline-gated currents are inhibited by alpha-bungarotoxin. To examine whether PGE(2) modulates nAChRs at synaptic sites, we studied the effects of PGE(2) on amplitude and decay of synaptic currents in visceral motoneuron-sympathetic neuron co-cultures. PGE(2) significantly decreases the amplitude of miniature excitatory postsynaptic currents (mEPSCs), consistent with the predominant inhibition by PGE(2) of all but the 23 pS subtype. The time constant of mEPSCs at PGE(2)-treated synapses is prolonged, which is also consistent with an increased contribution of the longer open duration of the 23 pS nAChR subtype with PGE(2) treatment. To examine the presynaptic effect of PGE(2), nanomolar nicotine was used. Nicotine induces facilitation of synaptic transmission by increasing mEPSC frequency, an action thought to involve presynaptic, alpha 7-containing nAChRs. In the presence of PGE(2), nicotine-induced synaptic facilitation persists. Thus the net effect of PGE(2) is to alter the profile of nAChRs contributing to synaptic transmission from larger conductance, briefer opening channels to smaller conductance, longer opening events. This subtype-specific modulation of nAChRs by PGE(2) may provide a mechanism for selective activation and suppression of synaptic pathways mediated by different nAChR subtype(s) at both pre- and postsynaptic sites. PMID- 11387398 TI - Comparison of the effect of intrathecal administration of clonidine and yohimbine on the locomotion of intact and spinal cats. AB - Several studies have shown that noradrenergic mechanisms are important for locomotion. For instance, L-dihydroxyphenylalanine (L-DOPA) can initiate "fictive" locomotion in immobilized acutely spinalized cats and alpha(2) noradrenergic agonists, such as 2,6,-dichloro-N-2-imidazolidinylid-enebenzenamine (clonidine), can induce treadmill locomotion soon after spinalization. However, the activation of noradrenergic receptors may be not essential for the basic locomotor rhythmicity because chronic spinal cats can walk with the hindlimbs on a treadmill in the absence of noradrenergic stimulation because the descending pathways are completely severed. This suggests that locomotion, in intact and spinal conditions, is probably expressed and controlled through different neurotransmitter mechanisms. To test this hypothesis, we compared the effect of the alpha(2) agonist, clonidine, and the antagonist (16 alpha, 17 alpha)-17 hydroxy yohimbine-16-carboxylic acid methyl ester hydrochloride (yohimbine), injected intrathecally at L(3)--L(4) before and after spinalization in the same cats chronically implanted with electrodes to record electromyograms (EMGs). In intact cats, clonidine (50-150 microg/100 microl) modulated the locomotor pattern slightly causing a decrease in duration of the step cycle accompanied with some variation of EMG burst amplitude and duration. In the spinal state, clonidine could trigger robust and sustained hind limb locomotion in the first week after the spinalization at a time when the cats were paraplegic. Later, after the spontaneous recovery of a stable locomotor pattern, clonidine prolonged the cycle duration, increased the amplitude and duration of flexor and extensor bursts, and augmented the foot drag at the onset of swing. In intact cats, yohimbine at high doses (800--1600 microg/100 microl) caused major walking difficulties characterized by asymmetric stepping, stumbling with poor lateral stability, and, at smaller doses (400 microg/100 microl), only had slight effects such as abduction of one of the hindlimbs and the turning of the hindquarters to one side. After spinalization, yohimbine had no effect even at the largest doses. These results indicate that, in the intact state, noradrenergic mechanisms probably play an important role in the control of locomotion since blocking the receptors results in a marked disruption of walking. In the spinal state, although the receptors are still present and functional since they can be activated by clonidine, they are seemingly not critical for the spontaneous expression of spinal locomotion since their blockade by yohimbine does not impair spinal locomotion. It is postulated therefore that the expression of spinal locomotion must depend on the activation of other types of receptors, probably related to excitatory amino acids. PMID- 11387397 TI - Presynaptic group II mGluR inhibition of short-term depression in the medial perforant path of the dentate gyrus in vitro. AB - Inhibition of short-term plasticity by activation of presynaptic group II metabotropic glutamate receptors (group II mGluR) was investigated in the medial perforant path of the dentate gyrus in the hippocampus in vitro. Brief trains of stimulation (10 stimuli at 1--200 Hz) evoked short-term depression of field excitatory postsynaptic potentials (EPSPs). The steady-state level of depression, measured after 10 stimuli, was frequency dependent, increasing between 1 and 200 Hz. Activation of group II mGluR by the selective agonist LY354740 did not alter short-term depression evoked by frequencies up to 10 Hz, but did inhibit short term depression evoked at higher frequencies in a frequency- and concentration dependent manner. The time-averaged postsynaptic response (EPSP per unit time) was found to increase linearly with frequency up to approximately 20 Hz. At higher frequencies, the response plateaued, thereby becoming independent of frequency. Frequencies above this were differentiated only during the transient postsynaptic response that accompanies changes in firing rates. Activation of presynaptically located group II mGluR increased the frequency at which the EPSP per unit time plateaued up to 30-50 Hz. PMID- 11387399 TI - Odor-stimulated phosphatidylinositol 3-kinase in lobster olfactory receptor cells. AB - Two antagonists of phosphoinositide 3-OH kinases (PI3Ks), LY294002 and Wortmannin, reduced the magnitude of the receptor potential in lobster olfactory receptor neurons (ORNs) recorded by patch clamping the cells in vivo. An antibody directed against the c-terminus of human PI3K-P110 beta detected a molecule of predicted size in the outer dendrites of the ORNs. Two 3-phosphoinositides, PI(3,4)P(2) (1--4 microM) and PI(3,4,5)P(3) (1--4 microM) applied to the cytoplasmic side of inside-out patches taken from cultured lobster ORNs, reversibly activated a Na(+)-gated channel previously implicated in the transduction cascade in these cells. 3-Phosphoinositides were the most effective phosphoinositide (1 microM) in enhancing the open probability of the channel. Collectively, these results implicate 3-phosphoinositides in lobster olfactory transduction and raise the need to consider the 3-phosphoinositide pathway in olfactory transduction. PMID- 11387400 TI - Progression in neuronal processing for saccadic eye movements from parietal cortex area lip to superior colliculus. AB - Neurons in both the lateral intraparietal area (LIP) of the monkey parietal cortex and the intermediate layers of the superior colliculus (SC) are activated well in advance of the initiation of saccadic eye movements. To determine whether there is a progression in the covert processing for saccades from area LIP to SC, we systematically compared the discharge properties of LIP output neurons identified by antidromic activation with those of SC neurons collected from the same monkeys. First, we compared activity patterns during a delayed saccade task and found that LIP and SC neurons showed an extensive overlap in their responses to visual stimuli and in their sustained activity during the delay period. The saccade activity of LIP neurons was, however, remarkably weaker than that of SC neurons and never occurred without any preceding delay activity. Second, we assessed the dependence of LIP and SC activity on the presence of a visual stimulus by contrasting their activity in delayed saccade trials in which the presentation of the visual stimulus was either sustained (visual trials) or brief (memory trials). Both the delay and the presaccadic activity levels of the LIP neuronal sample significantly depended on the sustained presence of the visual stimulus, whereas those of the SC neuronal sample did not. Third, we examined how the LIP and SC delay activity relates to the future production of a saccade using a delayed GO/NOGO saccade task, in which a change in color of the fixation stimulus instructed the monkey either to make a saccade to a peripheral visual stimulus or to withhold its response and maintain fixation. The average delay activity of both LIP and SC neuronal samples significantly increased by the advance instruction to make a saccade, but LIP neurons were significantly less dependent on the response instruction than SC neurons, and only a minority of LIP neurons was significantly modulated. Thus despite some overlap in their discharge properties, the neurons in the SC intermediate layers showed a greater independence from sustained visual stimulation and a tighter relationship to the production of an impending saccade than the LIP neurons supplying inputs to the SC. Rather than representing the transmission of one processing stage in parietal cortex area LIP to a subsequent processing stage in SC, the differences in neuronal activity that we observed suggest instead a progressive evolution in the neuronal processing for saccades. PMID- 11387401 TI - Stimulation of Na-K-2Cl cotransporter in neurons by activation of Non-NMDA ionotropic receptor and group-I mGluRs. AB - In a previous study, we found that Na(+)-K(+)-2Cl(-) cotransporter in immature cortical neurons was stimulated by activation of the ionotropic N-methyl-D aspartate (NMDA) glutamate receptor in a Ca(2+)-dependent manner. In this report, we investigated whether the Na(+)-K(+)-2Cl(-) cotransporter in immature cortical neurons is stimulated by non-NMDA glutamate receptor-mediated signaling pathways. Expression of the Na(+)-K(+)-2Cl(-) cotransporter and metabotropic glutamate receptors (mGluR1 and 5) was detected in cortical neurons via immunoblotting and immunofluorescence staining. Significant stimulation of cotransporter activity was observed in the presence of both trans-(+/-)-1-aminocyclopentane-trans-1,3 dicarboxylic acid (trans-ACPD) (10 microM), a metabotropic glutamate receptor (mGluR) agonist, and (RS)-3,5-dihydroxyphenylglycine (DHPG) (20 microM), a selective group-I mGluR agonist. Both trans-ACPD and DHPG-mediated effects on the cotransporter were eradicated by bis-(o-aminophenoxy)-N,N,N',N'-tetraacetic acid AM, a Ca(2+) chelator. In addition, DHPG-induced stimulation of the cotransporter activity was inhibited in the presence of mGluRs antagonist (RS)-1-aminoindan-1,5 dicarboxylic acid (AIDA) (1 mM) and also with selective mGluR1 antagonist 7 (hydroxyimino)cyclopropa[b]chromen-1a-carboxylate ethyl ester (CPCCOEt) (100 microM). A DHPG-induced rise in intracellular Ca(2+) in cortical neurons was detected with Fura-2. Moreover, DHPG-mediated stimulation of the cotransporter was abolished by inhibition of Ca(2+)/CaM kinase II. Interestingly, the cotransporter activity was increased by activation of alpha-amino-3-hydroxy-5 methyl-4-isoxazolepropionic acid (AMPA) receptor. These results suggest that the Na(+)-K(+)-2Cl(-) cotransporter in immature cortical neurons is stimulated by group-I mGluR- and AMPA-mediated signal transduction pathways. The effects are dependent on a rise of intracellular Ca(2+). PMID- 11387402 TI - On the relationship between joint angular velocity and motor cortical discharge during reaching. AB - Single-unit activity in area M1 was recorded in awake, behaving monkeys during a three-dimensional (3D) reaching task performed in a virtual reality environment. This study compares motor cortical discharge rate to both the hand's velocity and the arm's joint angular velocities. Hand velocity is considered a parameter of extrinsic space because it is measured in the Cartesian coordinate system of the monkey's workspace. Joint angular velocity is considered a parameter of intrinsic space because it is measured relative to adjacent arm/body segments. In the initial analysis, velocity was measured as the difference in hand position or joint posture between the beginning and ending of the reach. Cortical discharge rate was taken as the mean activity between these two times. This discharge rate was compared through a regression analysis to either an extrinsic-coordinate model based on the three components of hand velocity or to an intrinsic coordinate model based on seven joint angular velocities. The model showed that velocities about four degrees-of-freedom (elbow flexion/extension, shoulder flexion/extension, shoulder internal/external rotation, and shoulder adduction/abduction) were those best represented in the sampled population of recorded activity. Patterns of activity recorded across the cortical population at each point in time throughout the task were used in a second analysis to predict the temporal profiles of joint angular velocity and hand velocity. The population of cortical units from area M1 matched the hand velocity and three of the four major joint angular velocities. However, shoulder adduction/abduction could not be predicted even though individual cells showed good correlation to movement on this axis. This was also the only major degree-of-freedom not well correlated to hand velocity, suggesting that the other apparent relations between joint angular velocity and neuronal activity may be due to intrinsic-extrinsic correlations inherent in reaching movements. PMID- 11387403 TI - Primate rhinal cortex participates in both visual recognition and working memory tasks: functional mapping with 2-DG. AB - The rhinal cortex in the medial temporal lobe has been implicated in object recognition memory tasks and indeed is considered to be the critical node in a visual memory network. Previous studies using the 2-deoxyglucose method have shown that thalamic and hippocampal structures thought to be involved in visual recognition memory are also engaged by spatial and object working memory tasks in the nonhuman primate. Networks engaged in memory processing can be recognized by analysis of patterns of activation accompanying performance of specifically designed tasks. In the present study, we compared metabolic activation of the entorhinal and perirhinal cortex during the performance of three working memory tasks [delayed response (DR), delayed alternation (DA), and delayed object alternation (DOA)] to that induced by a standard recognition memory task [delayed match-to-sample (DMS)] and a sensorimotor control task in rhesus monkeys. A region-of-interest analysis revealed elevated local cerebral glucose utilization in the perirhinal cortex in animals performing the DA, DOA, and DMS tasks, and animals performing the DMS task were distinct in showing a strong focus of activation in the lateral perirhinal cortex. No significant differences were evident between groups performing memory and control tasks in the entorhinal cortex. These findings suggest that the perirhinal cortex may play a much broader role in memory processing than has been previously thought, encompassing explicit working memory as well as recognition memory. PMID- 11387404 TI - Hemispheric lateralization of somatosensory processing. AB - Processing of both painful and nonpainful somatosensory information is generally thought to be subserved by brain regions predominantly contralateral to the stimulated body region. However, lesions to right, but not left, posterior parietal cortex have been reported to produce a unilateral tactile neglect syndrome, suggesting that components of somatosensory information are preferentially processed in the right half of the brain. To better characterize right hemispheric lateralization of somatosensory processing, H(2)(15)O positron emission tomography (PET) of cerebral blood flow was used to map brain activation produced by contact thermal stimulation of both the left and right arms of right handed subjects. To allow direct assessment of the lateralization of activation, left- and right-sided stimuli were delivered during separate PET scans. Both innocuous (35 degrees C) and painful (49 degrees C) stimuli were employed to determine whether lateralized processing occurred in a manner related to perceived pain intensity. Subjects were also scanned during a nonstimulated rest condition to characterize activation that was not related to perceived pain intensity. Pain intensity-dependent and -independent changes in activation were identified in separate multiple regression analyses. Regardless of the side of stimulation, pain intensity--dependent activation was localized to contralateral regions of the primary somatosensory cortex, secondary somatosensory cortex, insular cortex, and bilateral regions of the cerebellum, putamen, thalamus, anterior cingulate cortex, and frontal operculum. No hemispheric lateralization of pain intensity-dependent processing was detected. In sharp contrast, portions of the thalamus, inferior parietal cortex (BA 40), dorsolateral prefrontal cortex (BA 9/46), and dorsal frontal cortex (BA 6) exhibited right lateralized activation during both innocuous and painful stimulation, regardless of the side of stimulation. Thus components of information arising from the body surface are processed, in part, by right lateralized systems analogous to those that process auditory and visual spatial information arising from extrapersonal space. Such right lateralized processing can account for the left somatosensory neglect arising from injury to brain regions within the right cerebral hemisphere. PMID- 11387405 TI - Differential fronto-parietal activation depending on force used in a precision grip task: an fMRI study. AB - Recent functional magnetic resonance imaging (fMRI) studies suggest that the control of fingertip forces between the index finger and the thumb (precision grips) is dependent on bilateral frontal and parietal regions in addition to the primary motor cortex contralateral to the grasping hand. Here we use fMRI to examine the hypothesis that some of the areas of the brain associated with precision grips are more strongly engaged when subjects generate small grip forces than when they employ large grip forces. Subjects grasped a stationary object using a precision grip and employed a small force (3.8 N) that was representative of the forces that are typically used when manipulating small objects with precision grips in everyday situations or a large force (16.6 N) that represents a somewhat excessive force compared with normal everyday usage. Both force conditions involved the generation of time-variant static and dynamic grip forces under isometric conditions guided by auditory and tactile cues. The main finding was that we observed stronger activity in the bilateral cortex lining the inferior part of the precentral sulcus (area 44/ventral premotor cortex), the rostral cingulate motor area, and the right intraparietal cortex when subjects applied a small force in comparison to when they generated a larger force. This observation suggests that secondary sensorimotor related areas in the frontal and parietal lobes play an important role in the control of fine precision grip forces in the range typically used for the manipulation of small objects. PMID- 11387406 TI - Cerebral blood-flow changes induced by paired-pulse transcranial magnetic stimulation of the primary motor cortex. AB - Positron emission tomography (PET) was used to assess changes in regional cerebral blood flow (CBF) induced by paired-pulse transcranial magnetic stimulation (TMS) of primary motor cortex (M1). The study was performed in eight normal volunteers using two Magstim-200 stimulators linked with a Bistim module. A circular TMS coil was held in the scanner by a mechanical arm and located over the left M1. Surface electrodes were used to record motor evoked potentials (MEPs) from the contralateral first dorsal interosseous muscle (FDI). Cortical excitability was evaluated in the relaxed FDI using a paired conditioning-test stimulus paradigm with two interstimulus intervals (ISIs): 3 and 12 ms. The subjects were scanned three times during each of the following four conditions: 1) baseline with no TMS (BASE); 2) single-pulse TMS (TMSsing); 3) 3-ms paired pulse TMS (TMS3); and 4) 12-ms paired-pulse TMS (TMS12). CBF and peak-to-peak MEP amplitudes were measured over each 60-s scanning period. To assess TMS-induced changes in CBF, a t-statistic map was generated by first subtracting the single pulse TMS condition from the 3- and 12-ms paired-pulse TMS conditions and then correlating the CBF differences, respectively, with the amount of suppression and facilitation of the EMG responses. A significant positive correlation was observed between the CBF difference (TMS3-TMSsing) and the amount of suppression of EMG response, as well as between the CBF difference (TMS12-TMSsing) and the amount of facilitation of EMG response. This positive correlation was observed in the left M1, left lateral premotor cortex, and right M1 in the case of 3-ms paired-pulse TMS, but only in the left M1 in the case of 12-ms paired-pulse TMS. The above pattern of CBF response to paired-pulse TMS supports the possibility that suppression and facilitation of the EMG response are mediated by different populations of cortical interneurons. PMID- 11387407 TI - Ankle muscle stiffness in the control of balance during quiet standing. AB - This research presents new data and reanalyzed information to refute the criticisms of our model of stiffness control during quiet standing. A re-review of their references to biomechanical research on muscle ankle stiffness confirmed muscle stiffness estimates of the ankle series elastic elements that agreed closely with our estimates. A new technique is presented that directly estimates the muscle stiffness from the ankle moment (N. m) and sway angle (deg). The linear regression of 10 subjects standing quietly for 10 s estimated the stiffness (N x m/deg) to be safely above the gravitational spring. The R(2) scores for this linear regression averaged 0.92, confirming how closely the model approached a perfect spring that would have an R(2) = 1. These results confirm our model of a simple muscle stiffness control and refutes the criticisms. PMID- 11387408 TI - Serotonin and NO complementarily regulate generation of oscillatory activity in the olfactory CNS of a terrestrial mollusk. AB - Synchronous oscillation of membrane potentials, generated by assemblies of neurons, is a prominent feature in the olfactory systems of many vertebrate and invertebrate species. However, its generation mechanism is still controversial. Biogenic amines play important roles for mammalian olfactory learning and are also implicated in molluscan olfactory learning. Here, we investigated the role of serotonin, a biogenic amine, in the oscillatory dynamics in the procerebrum (PC), the molluscan olfactory center. Serotonin receptor blockers inhibited the spontaneous synchronous oscillatory activity of low frequency (approximately 0.5 Hz) in the PC. This was due to diminishing the periodic slow oscillation of membrane potential in bursting (B) neurons, which are essential neuronal elements for the synchronous oscillation in the PC. On the other hand, serotonin enhanced the amplitude of the slow oscillation in B neurons and subsequently increased the number of spikes in each oscillatory cycle. These results show that the extracellular serotonin level regulates the oscillation amplitude in B neurons and thus serotonin may be called an oscillation generator in the PC. Although nitric oxide (NO) is known to also be a crucial factor for generating the PC oscillatory activity and setting the PC oscillation frequency, the present study showed that NO only regulates the oscillation frequency in B neurons but could not increase the spikes in each oscillatory cycle. These results suggest complementary regulation of the PC oscillatory activity: NO determines the probability of occurrence of slow potentials in B neurons, whereas serotonin regulates the amplitude in each cycle of the oscillatory activity in B neurons. PMID- 11387409 TI - Disynaptic inhibition of omnipause neurons following electrical stimulation of the superior colliculus in alert cats. AB - We investigated the synaptic organization responsible for the inhibition of omnipause neurons (OPNs) following stimulation of the superior colliculus (SC) in alert cats. Stimulation electrodes were implanted bilaterally in the rostral and caudal SC where a short-pulse train induced small and large saccades, respectively. Effects of single-pulse stimulation on OPNs were examined with intracellular and extracellular recordings. In contrast to monosynaptic excitatory postsynaptic potentials, which were induced by rostral SC stimulation, inhibitory postsynaptic potentials were induced with disynaptic latencies (1.3- 1.9 ms) from both the rostral and caudal SC in most OPNs. Analysis of a larger extracellular sample complemented intracellular observations. Monosynaptic activation of OPNs was elicited more frequently from rostral sites than from caudal sites, whereas spike suppression with disynaptic latencies was induced by caudal as well as rostral stimulation with similar frequencies. The results imply that disynaptic inhibition is produced by activation of SC cells that are distributed over wide regions related to saccades of different sizes. We suggest that signals from these neurons initiate a saccadic pause of OPNs through single inhibitory interneurons. PMID- 11387410 TI - Postlesional vestibular reorganization in frogs: evidence for a basic reaction pattern after nerve injury. AB - Nerve injury induces a reorganization of subcortical and cortical sensory or motor maps in mammals. A similar process, vestibular plasticity 2 mo after unilateral section of the ramus anterior of N. VIII was examined in this study in adult frogs. The brain was isolated with the branches of both N. VIII attached. Monosynaptic afferent responses were recorded in the vestibular nuclei on the operated side following ipsilateral electric stimulation either of the sectioned ramus anterior of N. VIII or of the intact posterior vertical canal nerve. Excitatory and inhibitory commissural responses were evoked by separate stimulation of each of the contralateral canal nerves in second-order vestibular neurons. The afferent and commissural responses of posterior vertical canal neurons recorded on the operated side were not altered. However, posterior canal related afferent inputs had expanded onto part of the deprived ramus anterior neurons. Inhibitory commissural responses evoked from canal nerves on the intact side were detected in significantly fewer deprived ramus anterior neurons than in controls, but excitatory commissural inputs from the three contralateral canal nerves had expanded. This reactivation might facilitate the survival of deprived neurons and reduce the asymmetry in bilateral resting activities but implies a deterioration of the original spatial response tuning. Extensive similarities at the synaptic and network level were noted between this vestibular reorganization and the postlesional cortical and subcortical reorganization of sensory representations in mammals. We therefore suggest that nerve injury activates a fundamental neural reaction pattern that is common between sensory modalities and vertebrate species. PMID- 11387412 TI - Foot and mouth disease: why not vaccinate? PMID- 11387411 TI - Drug-resistant tuberculosis. PMID- 11387413 TI - Water retention and aquaporins in heart failure, liver disease and pregnancy. PMID- 11387414 TI - Carbon monoxide poisoning. PMID- 11387415 TI - Chlamydial infection in sheep: immune control versus fetal pathology. PMID- 11387416 TI - Hand hygiene--the case for evidence-based education. PMID- 11387417 TI - Design or accident? The natural history of teenage pregnancy. AB - The UK has the highest rate of teenage pregnancy in Western Europe. A retrospective record-based study was conducted in an East Devon general practice to gain greater understanding of the outcome of first teenage pregnancy and subsequent reproductive history. The comparison group was women who had first conceived between the ages of 25 and 29 years. 149/673 women born between 1968 and 1977 became pregnant when teenagers. Of these, 70 (47%) had the baby, 67 (45%) had a termination and 10 (7%) had a spontaneous miscarriage; 2 others experienced fetal loss. Of the women aged 25-29 at first conception, 127 (92%) had the baby, 6 (4%) had a termination and 5 (4%) had a miscarriage. 40 (27%) of the teenage group went on to have a second teenage pregnancy, including 12 of the 67 who had their first pregnancy terminated. Although teenage pregnancy is often viewed as unplanned and unwanted, the reality is more complex. Among this group, many first pregnancies were desired. Even among those whose first pregnancy was terminated, 18% went on to have a baby while still a teenager. PMID- 11387419 TI - An audit of audits: are we completing the cycle? AB - Clinical audit plays an important part in the drive to improve quality of patient care and thus forms a cornerstone of clinical governance. We evaluated the standard of clinical audits conducted by all departments in a teaching hospital between 1996 and 1997. Of a total of 213 audits carried out, 102 (48%) were 'partial' and only 29 (14%) were 'full'. Recommendations for improvement emerged from 134 (63%) of the audits performed. In only 51 audits (24%) was the cycle completed by re-auditing, during the subsequent 3 years. Most departments undertake clinical audits but failure to close the loop undermines their effectiveness and wastes resources. PMID- 11387418 TI - A diagnostic sign in migraine? AB - At the bedside it was noted that, after ocular fundoscopy, patients with migraine complained more often of an after-image than did non-migraineurs. This phenomenon was then investigated in consecutive patients attending a general neurology outpatient clinic. The relative risk for the diagnosis of migraine in patients reporting an after-image was 2.91 (95% confidence interval 1.96 to 4.34), and the sensitivity, specificity and positive predictive value of this observation for the diagnosis of migraine were 0.63, 0.75 and 0.55 respectively. After-images were equally likely to be reported by migraineurs with and without aura, and by patients with migraine equivalents. The after-image phenomenon probably reflects the heightened sensitivity to visual stimuli of patients with migraine. Although a diagnosis of migraine is primarily established by the patient's history, the presence of an after-image following ocular fundoscopy may support this diagnosis. PMID- 11387420 TI - Off-the-cuff cellular phone consultations in a family practice. AB - In Israel the public tends to make use of informal medicine alongside organized health services, and cellular phones now allow contact with physicians at almost any time or place. For three months in 1999 a family physician documented all consultations on medical subjects conducted by cellular phone, the phone being available 24 hours a day. There were 94 cellular phone medical consultations, of mean duration 5.8 min (range 2-18). Only 11 took place over the weekend, and 63 took place while the clinic was open. The most common reasons for consultation were advice on treatment (29%) and a second opinion (28%). In 48 cases the consultation was for a close relative rather than the caller. In 42 cases the request for consultation came while the physician was busy with other patients. The results of this small personal study confirm that the practice of informal consultations now extends to the cellular phone. Technologies of this sort demand new rules of conduct, if we are to avoid the various hazards of off-the-cuff medicine. PMID- 11387421 TI - Mondor meets Trendelenburg: penile vein thrombosis after varicose vein surgery. PMID- 11387422 TI - Reduced consciousness with a runny nose. PMID- 11387423 TI - Four children crushed in their driveways. PMID- 11387424 TI - Stiff-man syndrome in childhood. PMID- 11387425 TI - Bilateral hypoperfusion retinopathy. PMID- 11387426 TI - Gitelman's syndrome. PMID- 11387427 TI - Surgical treatment of cortical tremor. PMID- 11387429 TI - Spirituality in psychiatric education and training. PMID- 11387430 TI - Evidence-based art? PMID- 11387431 TI - Neurological complications of cervical spine manipulation. PMID- 11387432 TI - The Good Soldier Svejk syndrome. PMID- 11387433 TI - The Mozart effect. PMID- 11387434 TI - The Mozart effect. PMID- 11387435 TI - BCG immunotherapy for superficial bladder cancer. PMID- 11387436 TI - Mastectomy retaining nipple as well as areola. PMID- 11387437 TI - Experience before medical school. PMID- 11387438 TI - Changing relationship between the public and the medical profession. PMID- 11387439 TI - Whiplash injury. PMID- 11387440 TI - Stimulation of RNA polymerase II elongation by hepatitis delta antigen. AB - Transcription elongation by RNA polymerase II (RNAPII) is negatively regulated by the human factors DRB-sensitivity inducing factor (DSIF) and negative elongation factor (NELF). A 66-kilodalton subunit of NELF (NELF-A) shows limited sequence similarity to hepatitis delta antigen (HDAg), the viral protein required for replication of hepatitis delta virus (HDV). The host RNAPII has been implicated in HDV replication, but the detailed mechanism and the role of HDAg in this process are not understood. We show that HDAg binds RNAPII directly and stimulates transcription by displacing NELF and promoting RNAPII elongation. These results suggest that HDAg may regulate RNAPII elongation during both cellular messenger RNA synthesis and HDV RNA replication. PMID- 11387441 TI - Strange and unconventional isotope effects in ozone formation. AB - The puzzling mass-independent isotopic enrichment in ozone formation contrasts markedly with the more recently observed large unconventional mass-dependent ratios of the individual ozone formation rate constants in certain systems. An RRKM (Rice, Ramsperger, Kassel, Marcus)-based theory is used to treat both effects. Restrictions of symmetry on how energy is shared among the rotational/vibrational states of the ozone isotopomer, together with an analysis of the competition between the transition states of its two exit channels, permit the calculation of isotope effects consistent with a wide array of experimental results. PMID- 11387442 TI - Methylation of histone H4 at arginine 3 facilitating transcriptional activation by nuclear hormone receptor. AB - Acetylation of core histone tails plays a fundamental role in transcription regulation. In addition to acetylation, other posttranslational modifications, such as phosphorylation and methylation, occur in core histone tails. Here, we report the purification, molecular identification, and functional characterization of a histone H4-specific methyltransferase PRMT1, a protein arginine methyltransferase. PRMT1 specifically methylates arginine 3 (Arg 3) of H4 in vitro and in vivo. Methylation of Arg 3 by PRMT1 facilitates subsequent acetylation of H4 tails by p300. However, acetylation of H4 inhibits its methylation by PRMT1. Most important, a mutation in the S-adenosyl-l-methionine binding site of PRMT1 substantially crippled its nuclear receptor coactivator activity. Our finding reveals Arg 3 of H4 as a novel methylation site by PRMT1 and indicates that Arg 3 methylation plays an important role in transcriptional regulation. PMID- 11387443 TI - Can science save Africa? PMID- 11387444 TI - Ecoterrorism. Arson strikes research labs and tree farm in Pacific Northwest. PMID- 11387446 TI - Austrian universities. Scientists spar over reform plan. PMID- 11387445 TI - Paleontology. New dig at old trove yields giant sauropod. PMID- 11387447 TI - Space research. ESA embraces astrobiology. PMID- 11387448 TI - U.S. Congress. New leaders emerge after Senate shake-up. PMID- 11387449 TI - Ultrafast lasers. Strobe light breaks the attosecond barrier. AB - On page 1689 of this issue, physicists report creating the fastest strobe light ever made, with individual pulses lasting just 220 attoseconds, or 220 billionths of a billionth of a second. These unimaginably short pulses are the first to be confirmed as breaking the attosecond barrier, a goal of high-speed-laser researchers for nearly a decade. Such pulses may one day serve as an ultrafast camera, allowing researchers to freeze action and perhaps to spot the gyrations of individual electrons whirling around an atomic nucleus. PMID- 11387450 TI - Neurobiology. Bee dance reveals bee's-eye view. PMID- 11387451 TI - Astrophysics. Radical gravity theory hits large-scale snag. PMID- 11387452 TI - Astronomy. Deep-space 'filament' shows cosmic fabric. PMID- 11387453 TI - Astronomy. The most powerful action flick ever. PMID- 11387455 TI - Nuclear proliferation. New effort puts radiation sentinels at the borders. PMID- 11387454 TI - Nuclear proliferation. Nuclear trafficking: 'a real and dangerous threat'. PMID- 11387456 TI - Nuclear proliferation. Tracing the shadowy origins of nuclear contraband. PMID- 11387457 TI - Language, brain, and cognitive development meeting. What makes the mind dance and count. PMID- 11387458 TI - Infectious diseases. Is the U.S. doing enough to prevent mad cow disease? PMID- 11387459 TI - Infectious diseases. America's own prion disease? PMID- 11387460 TI - Essays on science and society. Morals and primordials. PMID- 11387461 TI - Cell biology. GGAs tie up the loose ends. PMID- 11387463 TI - Quantum computing. Computation from geometry. PMID- 11387462 TI - Microbiology. Cracking Listeria's password. PMID- 11387464 TI - Climate. Open windows to the polar oceans. PMID- 11387465 TI - Chemical physics. Single-molecule spectroscopy comes of age. PMID- 11387466 TI - Neuroimaging databases. AB - These are comments written by the Governing Council of the Organization for Human Brain Mapping (OHBM), the primary international organization dedicated to neuroimaging research. The purpose of these comments is to identify and frame issues concerning data sharing within the neuroimaging community. Data sharing has become an important issue in most fields of science. The neuroimaging community is no exception, and it clearly perceives potential benefits in such efforts, as have been realized in other fields such as genomics. At the same time, such efforts can be costly (both in time and expense), and there are important factors that differentiate brain imaging from other fields and that pose specific challenges to the generation of useful neuroimaging databases. These include the rapid pace of change in brain imaging technologies; the complexity of the variables that must be specified to meaningfully interpret the results (such as the method of image acquisition, behavioral design, and subject characteristics); and concerns about participant confidentiality. These issues are outlined with the goal of framing and promoting a public discussion of the benefits and risks of data sharing, which can inform the field of neuroimaging as well as others that face similar challenges. PMID- 11387467 TI - Observation of a train of attosecond pulses from high harmonic generation. AB - In principle, the temporal beating of superposed high harmonics obtained by focusing a femtosecond laser pulse in a gas jet can produce a train of very short intensity spikes, depending on the relative phases of the harmonics. We present a method to measure such phases through two-photon, two-color photoionization. We found that the harmonics are locked in phase and form a train of 250-attosecond pulses in the time domain. Harmonic generation may be a promising source for attosecond time-resolved measurements. PMID- 11387468 TI - Anomalous weak magnetism in superconducting YBa2Cu3O6+x. AB - For some time now, there has been considerable experimental and theoretical effort to understand the role of the normal-state "pseudogap" phase in underdoped high-temperature cuprate superconductors. Recent debate has centered on the question of whether the pseudogap is independent of superconductivity. We provide evidence from zero-field muon spin relaxation measurements in YBa2Cu3O6+x for the presence of small spontaneous static magnetic fields of electronic origin intimately related to the pseudogap transition. Our most significant finding is that, for optimal doping, these weak static magnetic fields appear well below the superconducting transition temperature. The two compositions measured suggest the existence of a quantum critical point somewhat above optimal doping. PMID- 11387469 TI - Geometric manipulation of trapped ions for quantum computation. AB - We propose an experimentally feasible scheme to achieve quantum computation based solely on geometric manipulations of a quantum system. The desired geometric operations are obtained by driving the quantum system to undergo appropriate adiabatic cyclic evolutions. Our implementation of the all-geometric quantum computation is based on laser manipulation of a set of trapped ions. An all geometric approach, apart from its fundamental interest, offers a possible method for robust quantum computation. PMID- 11387470 TI - Explaining the Weddell Polynya--a large ocean eddy shed at Maud Rise. AB - Satellite observations have shown the occasional occurrence of a large opening in the sea-ice cover of the Weddell Sea, Antarctica, a phenomenon known as the Weddell Polynya. The transient appearance, position, size, and shape of the polynya is explained here by a mechanism by which modest variations in the large scale oceanic flow past the Maud Rise seamount cause a horizontal cyclonic eddy to be shed from its northeast flank. The shed eddy transmits a divergent Ekman stress into the sea ice, leading to a crescent-shaped opening in the pack. Atmospheric thermodynamical interaction further enhances the opening by inducing oceanic convection. A sea-ice-ocean computer model simulation vividly demonstrates how this mechanism fully accounts for the characteristics that mark Weddell Polynya events. PMID- 11387471 TI - Photosynthesis-induced biofilm calcification and calcium concentrations in Phanerozoic oceans. AB - Photosynthetic carbon assimilation is commonly invoked as the cause of calcium carbonate precipitation in cyanobacterial biofilms that results in the formation of calcareous stromatolites. However, biofilm calcification patterns in recent lakes and simulation of photosynthetically induced rise in calcium carbonate supersaturation demonstrate that this mechanism applies only in settings low in dissolved inorganic carbon and high in calcium. Taking into account paleo-partial pressure curves for carbon dioxide, we show that Phanerozoic oceans sustaining calcified cyanobacteria must have had considerably higher calcium concentrations than oceans of today. In turn, the enigmatic lack of calcified cyanobacteria in stromatolite-bearing Precambrian sequences can now be explained as a result of high dissolved inorganic carbon concentrations. PMID- 11387472 TI - A giant sauropod dinosaur from an Upper Cretaceous mangrove deposit in Egypt. AB - We describe a giant titanosaurid sauropod dinosaur discovered in coastal deposits in the Upper Cretaceous Bahariya Formation of Egypt, a unit that has produced three Tyrannosaurus-sized theropods and numerous other vertebrate taxa. Paralititan stromeri is the first tetrapod reported from Bahariya since 1935. Its 1.69-meter-long humerus is longer than that of any known Cretaceous sauropod. The autochthonous scavenged skeleton was preserved in mangrove deposits, raising the possibility that titanosaurids and their predators habitually entered such environments. PMID- 11387473 TI - Climate-driven range expansion and morphological evolution in a marine gastropod. AB - Little is known about the phenotypic consequences of global climate change, despite the excellent Pleistocene fossil record of many taxa. We used morphological measurements from extant and Pleistocene populations of a marine gastropod (Acanthinucella spirata) in conjunction with mitochondrial DNA sequence variation from living populations to determine how populations responded phenotypically to Pleistocene climatic changes. Northern populations show little sequence variation as compared to southern populations, a pattern consistent with a recent northward range expansion. These recently recolonized northern populations also contain shell morphologies that are absent in extant southern populations and throughout the Pleistocene fossil record. Thus, contrary to traditional expectations that morphological evolution should occur largely within Pleistocene refugia, our data show that geographical range shifts in response to climatic change can lead to significant morphological evolution. PMID- 11387474 TI - Parent-offspring coadaptation and the dual genetic control of maternal care. AB - In many animal species, the amount of care provided by parents is determined through a complex interaction of offspring signals and responses by parents to those signals. As predicted by honest signaling theory, we show that in the burrower bug, Sehirus cinctus, maternal provisioning responds to experimental manipulations of offspring condition. Despite this predicted environmental influence, we find evidence from two cross-foster experiments that variation in maternal care also stems from two distinct genetic sources: variation among offspring in their ability to elicit care and variation among parents in their response to offspring signals. Furthermore, as predicted by maternal-offspring coadaptation theory, offspring signaling is negatively genetically correlated with maternal provisioning. PMID- 11387475 TI - Sorting of mannose 6-phosphate receptors mediated by the GGAs. AB - The delivery of soluble hydrolases to lysosomes is mediated by the cation independent and cation-dependent mannose 6-phosphate receptors. The cytosolic tails of both receptors contain acidic-cluster-dileucine signals that direct sorting from the trans-Golgi network to the endosomal-lysosomal system. We found that these signals bind to the VHS domain of the Golgi-localized, gamma-ear containing, ARF-binding proteins (GGAs). The receptors and the GGAs left the trans-Golgi network on the same tubulo-vesicular carriers. A dominant-negative GGA mutant blocked exit of the receptors from the trans-Golgi network. Thus, the GGAs appear to mediate sorting of the mannose 6-phosphate receptors at the trans Golgi network. PMID- 11387476 TI - Binding of GGA2 to the lysosomal enzyme sorting motif of the mannose 6-phosphate receptor. AB - The GGAs are a multidomain protein family implicated in protein trafficking between the Golgi and endosomes. Here, the VHS domain of GGA2 was shown to bind to the acidic cluster-dileucine motif in the cytoplasmic tail of the cation independent mannose 6-phosphate receptor (CI-MPR). Receptors with mutations in this motif were defective in lysosomal enzyme sorting. The hinge domain of GGA2 bound clathrin, suggesting that GGA2 could be a link between cargo molecules and clathrin-coated vesicle assembly. Thus, GGA2 binding to the CI-MPR is important for lysosomal enzyme targeting. PMID- 11387477 TI - G protein signaling from activated rat frizzled-1 to the beta-catenin-Lef-Tcf pathway. AB - The frizzled receptors, which mediate development and display seven hydrophobic, membrane-spanning segments, are cell membrane-localized. We constructed a chimeric receptor with the ligand-binding and transmembrane segments from the beta2-adrenergic receptor (beta2AR) and the cytoplasmic domains from rat Frizzled 1 (Rfz1). Stimulation of mouse F9 clones expressing the chimera (beta2AR-Rfz1) with the beta-adrenergic agonist isoproterenol stimulated stabilization of beta catenin, activation of a beta-catenin-sensitive promoter, and formation of primitive endoderm. The response was blocked by inactivation of pertussis toxin sensitive, heterotrimeric guanine nucleotide-binding proteins (G proteins) and by depletion of Galphaq and Galphao. Thus, G proteins are elements of Wnt/Frizzled-1 signaling to the beta-catenin-lymphoid-enhancer factor (LEF)-T cell factor (Tcf) pathway. PMID- 11387478 TI - A transgenic model for listeriosis: role of internalin in crossing the intestinal barrier. AB - Listeria monocytogenes is responsible for severe food-borne infections, but the mechanisms by which bacteria cross the intestinal barrier are unknown. Listeria monocytogenes expresses a surface protein, internalin, that interacts with a host receptor, E-cadherin, to promote entry into human epithelial cells. Murine E cadherin, in contrast to guinea pig E-cadherin, does not interact with internalin, excluding the mouse as a model for addressing internalin function in vivo. In guinea pigs and transgenic mice expressing human E-cadherin, internalin was found to mediate invasion of enterocytes and crossing of the intestinal barrier. These results illustrate how relevant animal models for human infections can be generated. PMID- 11387479 TI - Structure of complement receptor 2 in complex with its C3d ligand. AB - Complement receptor 2 (CR2/CD21) is an important receptor that amplifies B lymphocyte activation by bridging the innate and adaptive immune systems. CR2 ligands include complement C3d and Epstein-Barr virus glycoprotein 350/220. We describe the x-ray structure of this CR2 domain in complex with C3d at 2.0 angstroms. The structure reveals extensive main chain interactions between C3d and only one short consensus repeat (SCR) of CR2 and substantial SCR side-side packing. These results provide a detailed understanding of receptor-ligand interactions in this protein family and reveal potential target sites for molecular drug design. PMID- 11387480 TI - Insulin resistance and a diabetes mellitus-like syndrome in mice lacking the protein kinase Akt2 (PKB beta). AB - Glucose homeostasis depends on insulin responsiveness in target tissues, most importantly, muscle and liver. The critical initial steps in insulin action include phosphorylation of scaffolding proteins and activation of phosphatidylinositol 3-kinase. These early events lead to activation of the serine-threonine protein kinase Akt, also known as protein kinase B. We show that mice deficient in Akt2 are impaired in the ability of insulin to lower blood glucose because of defects in the action of the hormone on liver and skeletal muscle. These data establish Akt2 as an essential gene in the maintenance of normal glucose homeostasis. PMID- 11387481 TI - Genetics update : impact of the human genome projects and identification of a stroke gene. PMID- 11387482 TI - Ethical and methodological issues in pedigree stroke research. AB - BACKGROUND: Stroke is a complex genetic disorder with a variable phenotype. Investigations of heritable factors in complex genetic disorders use pedigree and genetic techniques, which pose different ethical and methodological challenges than those routinely encountered in therapeutic research. Building consensus on acceptable research practices in this field is vital to the success of multicentered collaborations. SUMMARY OF REVIEW: We review important ethical and methodological concerns related to the collection, storage, and release of pedigree research information. The human studies aspects of pedigree research are complicated methodologically because individuals can be active or passive participants and pedigrees can be proband derived, partially validated, or fully validated. Current research ethics frameworks do not work well outside of a dyadic researcher-subject relationship. Privacy and confidentiality for family members must be considered in pedigree research. Investigators should anticipate potential conflicts of interest among family members when designing a pedigree research protocol. CONCLUSIONS: We propose a "proband-initiated contact" methodology in which the proband or the proband's designate allows identification of potential families without breaching the privacy of individuals in the family. In situations in which family history data are collected without direct contact between researchers and individuals in the proband's family, an Institutional Review Board may waive consent by family members after appropriate review of the protocol and application of rules for granting waivers of consent. Certificates of Confidentiality should be considered. PMID- 11387483 TI - Low potentiality of angiotensin-converting enzyme gene insertion/deletion polymorphism as a useful predictive marker for carotid atherogenesis in a large general population of a Japanese city: the Suita study. AB - BACKGROUND AND PURPOSE: Some previous studies, almost all western, have investigated whether there is a relationship between the insertion/deletion (I/D) polymorphism of the angiotensin-converting enzyme (ACE) and carotid atherosclerosis. The results, however, have not been consistently positive. Further, there have been few investigations based on a large, general population. Therefore, the present study aimed to clarify whether ACE gene deletion polymorphism was associated with carotid atherosclerosis in a large Japanese general population with a more homogeneous genetic background than Caucasian populations. METHODS: Subjects aged 30 to 86 years were randomly selected from Suita City, located in Osaka, the second largest urban area of Japan, and included 1894 men and 2137 women. With the aid of high-resolution ultrasonography, carotid atherosclerosis was evaluated using our atherosclerotic indexes of intimal-medial thickness (IMT), plaque number (PN), plaque score (PS), and percentage of stenosis of the carotid artery assessed using high-resolution B mode ultrasonography. ACE gene I/D polymorphism was detected by polymerase chain reaction. RESULTS: There were no significant differences among the ACE genotypes for age and conventional cardiovascular risk factors, except for systolic blood pressure (SBP) and the percentage of hypertension in men. The values of IMT, PN, and PS as carotid atherosclerotic indexes were not significantly different among genotypes for either sex. After adjusting for age, body mass index, smoking habit, high-density lipoprotein cholesterol, triglycerides, presence of hypertension, presence of diabetes mellitus, and presence of hyperlipidemia, the estimated ORs for the presence of IMT >/=1.10 mm (defined as thickened IMT), according to ACE genotype (DD versus II, DD+ID versus II, and DD versus ID+II), for men were 0.80 (95% CI 0.60 to 1.23), 0.89 (0.62 to 1.29), and 0.89 (0.70 to 1.28), respectively. On the other hand, the ORs for women after the same adjustment were 0.92 (95% CI 0.58 to 1.35), 0.93 (0.59 to 1.45), and 0.91 (0.59 to 1.27), respectively. CONCLUSIONS: Our present data suggest that ACE I/D polymorphism is not potentially a useful predictive marker for carotid atherogenesis when investigated in a large and homogeneous general Japanese population of 4031 subjects, a finding similar to that in a Caucasian population study, the Perth Carotid Ultrasound Disease Assessment Study, an Australian study based on a general population using 1111 subjects. PMID- 11387484 TI - Chickenpox and stroke in childhood: a study of frequency and causation. AB - BACKGROUND AND PURPOSE: The purpose of this study was to determine whether infection with varicella is causal for arterial ischemic stroke (AIS) in children. METHODS: First, a prospective cohort study was conducted in young children (aged 6 months to 10 years) with AIS at 2 institutions (cohort study). The presence of varicella infection <12 months before AIS was determined and compared with the published frequency of varicella infection in the healthy pediatric population. The clinical and radiographic features of AIS were compared between the varicella and nonvaricella study cohorts. Second, a literature search of varicella-associated AIS was conducted, and the clinical and radiographic features were compared with the study nonvaricella cohort. RESULTS: In the cohort study, 22 (31%) of 70 consecutive children with AIS had a varicella infection in the preceding year compared with 9% in the healthy population. Children in the varicella cohort were more likely to have basal ganglia infarcts (P<0.001), abnormal cerebral vascular imaging (P<0.05), and recurrent AIS or transient ischemic attacks (P<0.05) than those in the nonvaricella cohort. The pooled literature analysis of 51 cases of varicella-associated AIS showed similar findings to the varicella cohort. CONCLUSION: In young children with AIS, there is a 3-fold increase in preceding varicella infection compared with published population rates, and varicella-associated AIS accounts for nearly one third of childhood AIS. Varicella-associated AIS has characteristic features, including a 2-fold increase in recurrent AIS and transient ischemic attacks. Varicella is an important risk factor for childhood AIS. PMID- 11387485 TI - Stress-induced blood pressure reactivity and incident stroke in middle-aged men. AB - BACKGROUND AND PURPOSE: Exaggerated blood pressure reactivity to stress is associated with atherosclerosis and hypertension, which are known stroke risk factors, but its relation to stroke is unknown. Previous work also indicates that the association between reactivity and cardiovascular diseases may be influenced by socioeconomic status. METHODS: The impact of blood pressure reactivity and socioeconomic status on incident stroke was examined in 2303 men (mean age, 52.8+/-5.1 years) from a population-based, longitudinal study of risk factors for ischemic heart disease in eastern FINLAND: Reactivity was calculated as the difference between blood pressure measured during the anticipatory phase of an exercise tolerance test (before exercise) and resting blood pressure, measured 1 week earlier. Mean systolic reactivity was 20 mm Hg (+/-15.9), and mean diastolic reactivity was 8.6 mm Hg (+/-8.5). Socioeconomic status was assessed as years of education. One hundred thirteen incident strokes (90 ischemic) occurred in 11.2 (+/-1.6) years of follow-up. RESULTS: Men with exaggerated systolic reactivity (>/=20 mm Hg) had 72% greater risk of any stroke (relative hazard ratio [RH], 1.72; 95% CI, 1.17 to 2.54) and 87% greater risk of ischemic stroke (RH, 1.87; 95% CI, 1.20 to 2.89) relative to less reactive men. Moreover, men who were high reactors and poorly educated were nearly 3 times more likely to suffer a stroke than better educated, less reactive men (RH, 2.90; 95% CI, 1.66 to 5.08). Adjustment for stroke risk factors had little impact on these associations. Diastolic reactivity was unrelated to stroke risk. CONCLUSIONS: Excessive sympathetic reactivity to stress may be etiologically important in stroke, especially ischemic strokes, and low socioeconomic status confers added risk. PMID- 11387486 TI - Sleep-related breathing disorders and risk of stroke. AB - BACKGROUND: Stroke and sleep-related breathing disorders are both common and are associated with significant morbidity and mortality. Several recent large epidemiological studies have shown a strong association between these 2 disorders independent of known risk factors for stroke. This article will outline the scientific basis for this relationship and suggest sleep-related breathing disorders as modifiable risk factors for stroke. SUMMARY OF REVIEW: Several studies have shown a characteristic circadian rhythmicity in stroke. We have discussed the influence of normal sleep states as well as the effect of sleep related breathing disorders on cerebral hemodynamics. The hemodynamic, metabolic, and hematologic changes during sleep-related breathing disorders in the form of decreased cerebral perfusion and increased coagulability are possible pathogenetic mechanisms for stroke. There are accumulating lines of evidence that sleep apnea disorder may indeed cause diurnal hypertension. However, the increased risk of stroke in patients with sleep-related breathing disorders appears to be independent of coexisting hypertension; the presence of hypertension would increase the risk even further. Furthermore, several studies have documented high prevalence of sleep apnea disorders in patients with transient ischemic attacks and stroke. CONCLUSIONS: Sleep-related breathing disorder appears to contribute as a risk factor for stroke through hemodynamic and hematologic changes. Because of the high prevalence of sleep apnea disorder in this population, patients with transient ischemic attacks and stroke should undergo evaluation for these disorders. PMID- 11387487 TI - Estimates of the prevalence of acute stroke impairments and disability in a multiethnic population. AB - BACKGROUND AND PURPOSE: The goals of the present study were to estimate the prevalence of acute impairments and disability in a multiethnic population of first-ever stroke and to identify differences in impairment and early disability between pathological and Bamford subtypes. Associations between impairments and death and disability at 3 months were identified. METHODS: Impairments that occur at the time of maximum neurological deficit were recorded, and disability according to the Barthel Index (BI) was assessed 1 week and 3 months after stroke in patients in the South London Stroke Register: RESULTS: Of 1259 registered patients, 6% had 1 or 2, 31.1% had 3 to 5, 50.6% had 6 to 10, and 10.6% had >10 impairments. Common impairments were weakness (upper limb, 77.4%), urinary incontinence (48.2%), impaired consciousness (44.7%), dysphagia (44.7%), and impaired cognition (43.9%). Patients with total anterior circulation infarcts had the highest age-adjusted prevalence of weakness, dysphagia, urinary incontinence, cognitive impairment, and disability. Patients with subarachnoid hemorrhage had the highest rates of coma. Patients with lacunar stroke had the high prevalence of weakness but were least affected by disability, incontinence, and cognitive dysfunction. Blacks had higher age- and sex-adjusted rates of disability in ischemic stroke (BI <20, odds ratio 2.76, 95% CI 1.47 to 5.21, P=0.002; BI <15, odds ratio 1.8, 95% CI 1.45 to 2.81, P=0.01) but impairment rates similar to those of whites. On multivariable analysis, incontinence, coma, dysphagia, cognitive impairment, and gaze paresis were independently associated with severe disability (BI <10) and death at 3 months. CONCLUSIONS: The extent of these findings indicates that an acute assessment of impairments and disability is necessary to determine the appropriate nursing and rehabilitation needs of patients with stroke. PMID- 11387488 TI - Greater Cincinnati/Northern Kentucky Stroke Study: volume of first-ever ischemic stroke among blacks in a population-based study. AB - BACKGROUND AND PURPOSE: The volume of ischemic stroke on CT scans has been studied in a standardized fashion in acute stroke therapy trials with median volumes between 10.5 to 55 cm(3). The volume of first-ever ischemic stroke in the population is not known. METHODS: The first phase of the population-based Greater Cincinnati/Northern Kentucky Stroke Study identified all ischemic strokes occurring in blacks in the greater Cincinnati region between January and June of 1993. The patients in this phase of the study who had a first-ever ischemic clinical stroke were identified, and the volume of ischemic stroke was measured. RESULTS: There were 257 verified clinical cases of ischemic stroke, of which 181 had a first-ever ischemic infarct. Imaging was available for 150 of these patients, and 79 had an infarct on the CT or MRI study that was definitely or possibly related to the clinical symptoms. For these patients, volumetric measurements were performed by means of the modified ellipsoid method. The median volume of first-ever ischemic stroke for the 79 patients was 2.5 cm(3) (interquartile range, 0.5 to 8.8 cm(3)). There was a significant relation between location of lesion and infarct size (P<0.001) and between volume and mechanism of stroke (P=0.001). CONCLUSIONS: The volume of first-ever ischemic stroke among blacks in our population-based study is smaller than has been previously reported in acute stroke therapy trials. The large proportion of small, mild strokes in blacks may be an important reason for the low percentage of patients who meet the inclusion criteria for tissue plasminogen activator. Further study is necessary to see if these results are generalizable to a multiracial population. PMID- 11387489 TI - Power transcranial Doppler ultrasound in the detection of intracranial aneurysms. AB - BACKGROUND AND PURPOSE: We sought to perform a large, prospective, multicenter, blinded study comparing power transcranial color duplex sonography (power TCDS) with intra-arterial digital subtraction angiography (IADSA) in the detection of intracranial aneurysms. METHODS: Contemporaneous TCDS and IADSA examinations were performed in 171 subjects with suspected intracranial aneurysm. Via the temporal bone window, a 2-dimensional hand-held noncontrast transcranial duplex ultrasound imaging system was used operating in power and spectral modes. Sonographers were blinded to clinical history and results of brain CT and IADSA. RESULTS: We found that 157 subjects (92%) had an adequate bone window. Sensitivity per patient was 0.78 (95% CI, 0.66 to 0.87) and 0.46 (95% CI, 0.36 to 0.56) for any anterior circulation aneurysms. Sensitivity was 0.35 (95% CI, 0.24 to 0.46) for aneurysms 5 mm. Accuracy was lower for aneurysms on the cavernous and terminal internal carotid arteries, including posterior communicating artery origin (0.71; 95% CI, 0.63 to 0.79), than for those on the anterior (0.82; 95% CI, 0.74 to 0.89) or the middle cerebral arteries (0.79; 95% CI, 0.71 to 0.86). CONCLUSIONS: Power TCDS is a promising, inexpensive, noninvasive test for anterior circulation intracranial aneurysms but is less sensitive per aneurysm than alternatives such as CT angiography or MR angiography. Sensitivity is poor for aneurysms 50% maximum) contraction of the target muscle. In 15 of 25 stroke patients, ipsilateral MEPs in the unaffected relaxed FDI (in biceps, 6 of 25 stroke patients) were evoked by stimulation of premotor areas of the affected hemisphere. Their latencies were shorter than those that MEPs evoked in the same muscle by stimulation of the motor cortex of the contralateral unaffected hemisphere. Such responses were never obtained in normal subjects and were mostly observed in patients with subcortical infarcts. Patients harboring these responses had slightly better bimanual dexterity after 6 months. CONCLUSIONS: Ipsilateral MEPs obtained in healthy individuals and stroke patients have different characteristics and probably different origins. In the former, they are probably conveyed via corticoreticulospinal or corticopropriospinal pathways, whereas in the latter, early ipsilateral MEPs could originate in hyperexcitable premotor areas. PMID- 11387492 TI - A modified National Institutes of Health Stroke Scale for use in stroke clinical trials: preliminary reliability and validity. AB - BACKGROUND AND PURPOSE: The National Institutes of Health Stroke Scale (NIHSS) is accepted widely for measuring acute stroke deficits in clinical trials, but it contains items that exhibit poor reliability or do not contribute meaningful information. To improve the scale for use in clinical research, we used formal clinimetric analyses to derive a modified version, the mNIHSS. We then sought to demonstrate the validity and reliability of the new mNIHSS. METHODS: The mNIHSS was derived from our prior clinimetric studies of the NIHSS by deleting poorly reproducible or redundant items (level of consciousness, face weakness, ataxia, dysarthria) and collapsing the sensory item into 2 responses. Reliability of the mNIHSS was assessed with the certification data originally collected to assess the reliability of investigators in the National Institute of Neurological Disorders and Stroke (NINDS) rtPA (recombinant tissue plasminogen activator) Stroke TRIAL: Validity of the mNIHSS was assessed with the outcome results of the NINDS rtPA Stroke Trial: RESULTS: Reliability was improved with the mNIHSS: the number of scale items with poor kappa coefficients on either of the certification tapes decreased from 8 (20%) to 3 (14%) with the mNIHSS. With the use of factor analysis, the structure underlying the mNIHSS was found identical to the original scale. On serial use of the scale, goodness of fit coefficients were higher with the mNIHSS. With data from part I of the trial data, the proportion of patients who improved >/=4 points within 24 hours after treatment was statistically significantly increased by tPA (odds ratio, 1.3; 95% confidence limits, 1.0, 1.8; P=0.05). Likewise, the odds ratio for complete/nearly complete resolution of stroke symptoms 3 months after treatment was 1.7 (95% confidence limits, 1.2, 2.6) with the mNIHSS. Other outcomes showed the same agreement when the mNIHSS was compared with the original scale. The mNIHSS showed good responsiveness, ie, was useful in differentiating patients likely to hemorrhage or have a good outcome after stroke. CONCLUSIONS: The mNIHSS appears to be identical clinimetrically to the original NIHSS when the same data are used for validation and reliability. Power appears to be greater with the mNIHSS with the use of 24 hour end points, suggesting the need for fewer patients in trials designed to detect treatment effects comparable to rtPA. The mNIHSS contains fewer items and might be simpler to use in clinical research trials. Prospective analysis of reliability and validity, with the use of an independently collected cohort, must be obtained before the mNIHSS is used in a research setting. PMID- 11387493 TI - A new rating scale for age-related white matter changes applicable to MRI and CT. AB - BACKGROUND AND PURPOSE: MRI is more sensitive than CT for detection of age related white matter changes (ARWMC). Most rating scales estimate the degree and distribution of ARWMC either on CT or on MRI, and they differ in many aspects. This makes it difficult to compare CT and MRI studies. To be able to study the evolution and possible effect of drug treatment on ARWMC in large patient samples, it is necessary to have a rating scale constructed for both MRI and CT. We have developed and evaluated a new scale and studied ARWMC in a large number of patients examined with both MRI and CT. METHODS: Seventy-seven patients with ARWMC on either CT or MRI were recruited and a complementary examination (MRI or CT) performed. The patients came from 4 centers in Europe, and the scans were rated by 4 raters on 1 occasion with the new ARWMC rating scale. The interrater reliability was evaluated by using kappa statistics. The degree and distribution of ARWMC in CT and MRI scans were compared in different brain areas. RESULTS: Interrater reliability was good for MRI (kappa=0.67) and moderate for CT (kappa=0.48). MRI was superior in detection of small ARWMC, whereas larger lesions were detected equally well with both CT and MRI. In the parieto-occipital and infratentorial areas, MRI detected significantly more ARWMC than did CT. In the frontal area and basal ganglia, no differences between modalities were found. When a fluid-attenuated inversion recovery sequence was used, MRI detected significantly more lesions than CT in frontal and parieto-occipital areas. No differences were found in basal ganglia and infratentorial areas. CONCLUSIONS: We present a new ARWMC scale applicable to both CT and MRI that has almost equal sensitivity, except for certain regions. The interrater reliability was slightly better for MRI, as was the detectability of small lesions. PMID- 11387494 TI - Acute stroke patterns in patients with internal carotid artery disease: a diffusion-weighted magnetic resonance imaging study. AB - BACKGROUND AND PURPOSE: Diffusion-weighted (DW) MRI is a sensitive method that facilitates early stroke pattern identification. There are limited data about the influence of stenosis grade on the development of particular stroke patterns in internal carotid artery (ICA) disease. We therefore investigated the lesion patterns on DW MRI in acute stroke patients with ICA disease. METHODS: DW MRI was analyzed in 102 consecutive acute stroke patients with different degrees of ipsilateral ICA disease. Patients were assigned to 1 of 5 observed lesion patterns: (1) territorial ischemia, (2) subcortical ischemia without or (3) with embolus fragmentation, (4) disseminated lesions in distal cortical regions, and (5) multiple lesions in hemodynamic risk zones. In addition, perfusion-weighted (PW) MRI and MR angiography information was included in the assessment. RESULTS: All patterns were observed in the different stages of ICA disease. Half of the patients with high-grade or subtotal stenosis had lesions in hemodynamic risk zones. Territorial stroke occurred in 47.6% of patients with ICA occlusion. Statistical analysis showed a significant relationship between the degree of stenosis and the observed stroke pattern (P=0.001). In 77.8% of patients exhibiting high-grade ICA stenosis, subtotal stenosis, or occlusion, the perfusion lesion was larger than the diffusion lesion (PW/DW mismatch). CONCLUSIONS: Although in the individual patient any of the infarct patterns may occur, in statistical terms the incidence of a particular stroke pattern is clearly dependent on the degree of stenosis. PMID- 11387495 TI - Hemorrhagic transformation of ischemic brain tissue: asymptomatic or symptomatic? AB - BACKGROUND AND PURPOSE: The term symptomatic hemorrhage secondary to ischemic stroke implies a clear causal relationship between clinical deterioration and hemorrhagic transformation (HT) regardless of the type of HT. The aim of this study was to assess which type of HT independently affects clinical outcome. METHODS: We used the data set of the European Cooperative Acute Stroke Study (ECASS) II for a post hoc analysis. All patients had a control CT scan after 24 to 96 hours or earlier in case of rapid and severe clinical deterioration. HT was categorized according to radiological criteria: hemorrhagic infarction type 1 and type 2 and parenchymal hematoma type 1 and type 2. The clinical course was prospectively documented with the National Institutes of Health Stroke Scale and the modified Rankin Scale: The independent risk of each type of HT was calculated for clinical deterioration at 24 hours and disability and death at 3 months after stroke onset and adjusted for possible confounding factors such as age, severity of stroke syndrome at baseline, and extent of the ischemic lesion on the initial CT. RESULTS: Compared with absence of HT, only parenchymal hematoma type 2 was associated with an increased risk for deterioration at 24 hours after stroke onset (adjusted odds ratio, 18; 95% CI, 6 to 56) and for death at 3 months (adjusted odds ratio, 11; 95% CI, 3.7 to 36). All other types of HT did not independently increase the risk of late deterioration. CONCLUSIONS: Only parenchymal hematoma type 2 independently causes clinical deterioration and impairs prognosis. It has a distinct radiological feature: it is a dense homogeneous hematoma >30% of the ischemic lesion volume with significant space occupying effect. PMID- 11387496 TI - Hemorrhagic transformation after fibrinolysis with tissue plasminogen activator: evaluation of role of hypertension with rat thromboembolic stroke model. AB - BACKGROUND AND PURPOSE: We used a rat model of thromboembolic stroke to evaluate whether hypertension increases the incidence of hemorrhage after fibrinolysis with tissue plasminogen activator (tPA). METHODS: In this model, a microclot suspension was injected into the middle cerebral artery territory to induce focal ischemia. Reperfusion was induced in spontaneously hypertensive rats (SHR) by administering tPA (10 mg/kg) intravenously at 2 hours or 6 hours after the onset of thromboembolic focal ischemia. In untreated control rats, saline was administered at 2 hours after ischemia. RESULTS: Hemorrhagic transformation was observed only in rats that received tPA at 6 hours (6 of 8 rats [75%]). Reduction of mean arterial blood pressure from 122+/-3 to 99+/-2 mm Hg with hydralazine, given to SHR for 1 week before ischemia, significantly decreased the incidence of hemorrhage in 2 of 11 rats (18%). tPA reduced infarct volumes, but cotreatment with hydralazine did not result in further protection. CONCLUSIONS: This study demonstrates that in this rat thromboembolic model of stroke, tPA-induced hemorrhage is dependent on blood pressure and that pharmacological reduction of hypertension during fibrinolysis can reduce the risk of hemorrhagic transformation. PMID- 11387497 TI - Rapid differential endogenous plasminogen activator expression after acute middle cerebral artery occlusion. AB - BACKGROUND AND PURPOSE: During focal cerebral ischemia, the microvascular matrix (ECM), which participates in microvascular integrity, is degraded and lost when neurons are injured. Loss of microvascular basal lamina antigens coincides with rapid expression of select matrix metalloproteinases (MMPs). Plasminogen activators (PAs) may also play a role in ECM degradation by the generation of plasmin or by MMP activation. METHODS: The endogenous expressions of tissue-type plasminogen activator (tPA), urokinase (uPA), and PA inhibitor-1 (PAI-1) were quantified in 10-microm frozen sections from ischemic and matched nonischemic basal ganglia and in the plasma of 34 male healthy nonhuman primates before and after middle cerebral artery occlusion (MCA:O). RESULTS: Within the ischemic basal ganglia, tissue uPA activity and antigen increased significantly within 1 hour after MCA:O (2P<0.005). tPA activity transiently decreased 2 hours after MCA:O (2P=0.01) in concert with an increase in PAI-1 antigen (2P=0.001) but otherwise did not change. The transient decrease in free tPA antigen was marked by an increase in the tPA-PAI-1 complex (2P<0.001). No significant relations to neuronal injury or intracerebral hemorrhage were discerned. CONCLUSIONS: The rapid increase in endogenous PA activity is mainly due to significant increases in uPA, but not tPA, within the ischemic basal ganglia after MCA:O. This increase and an increase in PAI-1 coincided with latent MMP-2 generation and microvascular ECM degeneration but not neuronal injury. PMID- 11387498 TI - Trends in acute ischemic stroke trials through the 20th century. AB - BACKGROUND AND PURPOSE: The advent of controlled clinical trials revolutionized clinical medicine over the course of the 20th century. The objective of this study was to quantitatively characterize developments in clinical trial methodology over time in the field of acute ischemic stroke. METHODS: All controlled trials targeting acute ischemic stroke with a final report in English were identified through MEDLINE and international trial registries. Data regarding trial design, implementation, and results were extracted. A formal 100 point scale was used to rate trial quality. RESULTS: A total of 178 controlled acute stroke trials were identified, encompassing 73 949 patients. Eighty-eight trials involved neuroprotective agents, 59 rheological/antithrombotic agents, 26 agents with both neuroprotective and rheological/antithrombotic effects, and 5 a nonpharmacological intervention. Only 3 trials met conventional criteria for a positive outcome. Between the 1950s and 1990s, the number of trials per decade increased from 3 to 99, and mean trial sample size increased from 38 (median, 26) to 661 (median, 113). During 1980-1999, median time window allowed for enrollment decreased per half decade from 48 to 12 hours. Reported pharmaceutical sponsorship increased substantially over time, from 38% before 1970 to 68% in the 1990s. Trial quality improved substantially from a median score of 12 in the 1950s to 72 in the 1990s. CONCLUSIONS: Accelerating trends in acute stroke controlled trials include growth in number, sample size, and quality, and reduction in entry time window. These changes reflect an increased understanding of the pathophysiology of acute stroke, the imperative for treatment initiation within a critical time window, and more sophisticated trial design. PMID- 11387499 TI - Barriers to acute stroke therapy and stroke prevention in Mexican Americans. AB - BACKGROUND AND PURPOSE: The purpose of this study was to identify specific targets to improve acute stroke treatment and stroke prevention in the Mexican American (MA) community. METHODS: A professional, academic survey research team provided structured questions and elicited responses from 719 subjects identified by random-digit dialing in the biethnic community of Corpus Christi, TEXAS: This community of approximately 300 000 is approximately half MA and half non-Hispanic white (NHW). The cooperation rate for the survey was 58%. RESULTS: MAs (n=357) were younger, less well educated, and had lower family income than NHWs (n=362, P=0.001). MAs had a higher prevalence of diabetes mellitus (P=0.001) but similar rates of hypertension, elevated cholesterol, and current tobacco use. MAs less commonly recognized that acute stroke therapy existed (P=0.029), were less likely to acknowledge a time window for acute stroke treatment (P=0.001), and were more reticent to say they would call 911 for stroke symptoms (P=0.01) than NHWS: MAs were significantly less able to recall stroke symptoms and risk factors than NHWS: Only approximately 20% of both groups identified stroke as the NO: 1 cause of disability. MAs expressed less confidence in their ability to prevent stroke (P<0.001), more distrust in the medical establishment (P=0.007), and more concern that money impedes their seeking medical care (P<0.001). CONCLUSIONS: There are significant barriers to both acute stroke treatment and stroke prevention in MAS: This study identifies specific targets amenable for testing in an intervention project following confirmation by a methodology other than telephone survey. PMID- 11387500 TI - Safety of intra-arterial thrombolysis in the postoperative period. AB - BACKGROUND AND PURPOSE: Limited systemic fibrinolysis and reduced dosage are features of intra-arterial thrombolyis (IAT) that may be advantageous in the treatment of postoperative strokes. However, IAT may increase the risk of surgical bleeding. We sought to determine the safety of postoperative IAT. METHODS: This was a retrospective case series from 6 university hospitals. All cases of IAT within 2 weeks of surgery were identified. Demographics, stroke mechanism, stroke severity, imaging and angiographic findings, time between surgery and lysis, thrombolytic agent used, surgical site bleeding, intracranial bleeding, and mortality rates were determined. Death or complications directly related to IAT were determined. RESULTS: Thirty-six patients (median age, 71.5 years; range, 45 to 85) were identified. Median time from surgery to stroke was 21.5 hours (range, 1 to 120). Open heart surgery was done in 18 (50%), carotid endarterectomy in 6 (17%), craniotomy in 3 (8%), ophthalmologic-ear, nose and throat surgery in 2 (6%), urologic-gynecologic surgery in 4 (11%), orthopedic surgery in 2 (6%), and plastic surgery in 1 (3%). The stroke causes were cardioembolism in 24 (67%), large-vessel atherosclerosis in 4 (11%), dissection in 3 (8%), postendarterectomy occlusion in 4 (11%), and radiation arteriopathy in 1 (3%). Median time to angiogram was 2.5 hours (0.1 to 5.5). Occlusion sites were M1 in 19 (53%), M2 in 9 (25%), internal carotid artery in 5 (14%), basilar artery in 2 (6%), and posterior communicating artery in 1 (3%). Thrombolysis was completed at a median of 4.5 hours (range, 1 to 8.0). Tissue plasminogen activator was used in 19 (53%) and urokinase in 17 (47%). Nine (26%) patients died. Surgical site bleeding occurred in 9 (25%) cases (minor in 6, major in 3). The major surgical bleeds were 2 post-craniotomy intracranial hemorrhages and 1 hemopericardium after coronary artery bypass grafting; all were fatal. Six deaths were non-IAT related: 3 caused by cerebral edema and 3 by systemic causes. Major bleeding complications were significantly more common among patients with craniotomy (P<0.02). CONCLUSIONS: Postoperative IAT carries a risk of bleeding in up to 25% of patients but is usually minor surgical site bleeding. Avoiding IAT in intracranial surgery patients may reduce complications. Mortality rate in this series was similar to that reported in prior IAT trials. IAT remains a viable therapeutic option for postoperative strokes. PMID- 11387501 TI - Variations between countries in outcome after stroke in the International Stroke Trial (IST). AB - BACKGROUND AND PURPOSE: This study describes the large variations in outcome after stroke between countries that participated in the International Stroke Trial and seeks to define whether they could be explained by variations in case mix or by other factors. METHODS: We analyzed data from the 15 116 patients recruited in Argentina, Australia, Italy, the Netherlands, Norway, Poland, Sweden, Switzerland, and the United Kingdom: We compared crude case fatality and the proportion of patients dead or dependent at 6 months; we used logistic regression to adjust for age, sex, atrial fibrillation, systolic blood pressure, level of consciousness, and number of neurological deficits. We used the frequency of prerandomization head CT scan and prescription of aspirin at discharge to indicate quality of care. RESULTS: The differences in outcome (all treatment groups combined) between the "best" and "worst" countries were very large for death (171 cases per 1000 patients) and for death or dependency (375 cases per 1000 patients). The differences were somewhat smaller after adjustment for case mix (160 and 311 cases per 1000 patients, respectively). Process of care may have accounted for some but not all of the residual variation in outcome. CONCLUSIONS: Adjustment for case mix explained only some of the variation in outcome between countries. The residual differences in outcome were too large to be explained by variations in care and most likely reflect differences in unmeasured baseline factors. These findings demonstrate the need to achieve balance of treatment and control within each country in multinational randomized controlled stroke trials and the need for caution in the interpretation of nonrandomized comparisons of outcome after stroke between countries. PMID- 11387502 TI - Neuroprotection in transient focal brain ischemia after delayed intravenous administration of brain-derived neurotrophic factor conjugated to a blood-brain barrier drug targeting system. AB - BACKGROUND AND PURPOSE: Neuroprotection with brain-derived neurotrophic factor (BDNF) requires direct injection into the brain owing to poor transport of the neurotrophin through the blood-brain barrier (BBB) in vivo. The present studies investigate whether BDNF alone or conjugated to a BBB drug targeting system is neuroprotective in focal, reversible brain ischemia after delayed intravenous administration at 60 or 120 minutes after middle cerebral arterial occlusion. METHODS: BDNF was conjugated to the OX26 murine monoclonal antibody to the rat transferrin receptor, which undergoes transport into brain from blood via the BBB transferrin receptor transcytosis system. After a 1-hour occlusion of the middle cerebral artery in nitrous oxide- ventilated animals with normal blood sugar, the brain was reperfused, and either BDNF or the BDNF/OX26 conjugate was administered as a single intravenous injection at a dose of 50 microg per rat. RESULTS: After the intravenous administration of unconjugated BDNF, there was no neuroprotection on the basis of analysis of brain at either 24 hours or 7 days after a 1-hour middle cerebral arterial occlusion. In contrast, there was a 68% and 70% reduction in cortical stroke volume at 24 hours and 7 days, respectively, after intravenous administration of 50 microg per rat of the BDNF conjugate (P<0.01). No effects on subcortical stroke volume were observed. CONCLUSIONS: These studies demonstrate marked neuroprotection in focal, transient brain ischemia with a single, delayed intravenous injection of BDNF if the neurotrophin is conjugated to a BBB drug targeting system. The neuroprotection is long lasting and persists for at least 7 days after a 1-hour middle cerebral artery occlusion. PMID- 11387503 TI - Clinical correlates and drug treatment of residents with stroke in long-term care. AB - BACKGROUND AND PURPOSE: Stroke incidence increases with age, and stroke survivors often require nursing home placement. Characteristics of these residents and factors associated with the secondary drug prevention of stroke in nursing homes have yet to be explored. METHODS: We used a population-based data set of all nursing home residents in 5 states (1992 to 1995). We identified 53 829 (20.4%) with a diagnosis of stroke on the Minimum Data Set assessment. We considered aspirin, dipyridamole, ticlopidine, or warfarin alone or in combination as secondary drug prevention. We used logistic regression modeling to identify independent predictors of drug treatment. RESULTS: Sixty-seven percent of stroke survivors were not receiving drug therapy for stroke prevention. Among those treated, most received aspirin alone (16%) or warfarin alone (10%). Independent predictors of drug treatment included comorbid conditions (eg, hypertension, atrial fibrillation, depression, Alzheimer's disease, dementia, gastrointestinal bleeding, and peptic ulcer disease). Those over the age of 85 years were less likely to be treated than those 65 to 74 years of age (odds ratio [OR], 0.86; 95% confidence interval [CI], 0.82 to 0.91); black residents were less likely to be treated than whites (OR, 0.80; 95% CI, 0.75 to 0.85); and those with severe cognitive (OR, 0.63; 95% CI, 0.60 to 0.67) or physical impairment (OR, 0.69; 95% CI, 0.64 to 0.75) were also less likely to receive drug treatment. CONCLUSIONS: Stroke is highly prevalent in long-term care. Despite the increased risk of subsequent stroke in the elderly, many are not being treated. The choice to treat or not to treat may be influenced by age, comorbidity, race/ethnicity, and cognitive or physical functioning. PMID- 11387504 TI - Role of endothelium in shear stress-induced constrictions in rat middle cerebral artery. AB - BACKGROUND AND PURPOSE: Luminal shear stress has been reported to constrict cerebral arteries and arterioles of several species. Although the endothelium is not required for this response, it is not known whether the endothelium enhances or attenuates shear stress-induced constrictions. METHODS: Middle cerebral arteries (MCAs) were isolated from male Long-Evans rats, mounted in a tissue bath, and pressurized to 80 mm Hg in the absence of luminal flow. In some MCAs, the endothelium was selectively loaded with fura 2 for the measurement of endothelial Ca(2+) concentration. Luminal shear stress was increased by adjusting luminal flow while maintaining a constant intraluminal pressure. RESULTS: After the development of spontaneous tone in MCAs without luminal flow, inside diameters were approximately 190 microm. MCAs constricted approximately 15% when luminal flow was increased to produce a shear stress of 50 dyne/cm(2). The shear stress-induced constrictions were more pronounced in vessels without intact endothelium. Scavenging reactive oxygen species with 4,5-dihydroxy-1,3-benzene disulfonic acid (Tiron) or superoxide dismutase/catalase significantly inhibited the shear stress-induced constrictions in vessels with intact endothelium and in vessels in which the endothelium had been removed. In intact vessels, endothelial Ca(2+) increased 33 nmol/L (from 133+/-11 to 166+/-12 nmol/L) when shear stress was increased to 50 dyne/cm(2). The presence of N(G)-nitro-L-arginine methyl ester (L-NAME), L-NAME+indomethacin, or L-NAME+indomethacin+charybdotoxin had no significant effect on the shear stress-induced constrictions in MCAs with intact endothelium. CONCLUSIONS: We conclude that the endothelium plays a role in attenuating the shear stress-induced constrictions in rat MCAS: The attenuation does not appear to be by release of NO, prostacyclin, or endothelium-derived hyperpolarizing factor. The endothelium apparently attenuates the constriction by an unknown dilating factor, by a dilating process, or simply by attenuating the mechanical force of the shear stress as it is transmitted to the abluminal side of the vessel. PMID- 11387505 TI - Early decrease in dna repair proteins, Ku70 and Ku86, and subsequent DNA fragmentation after transient focal cerebral ischemia in mice. AB - BACKGROUND AND PURPOSE: Ku70 and Ku86, multifunctional DNA repair proteins, bind to broken DNA ends, including double-strand breaks, and trigger a DNA repair pathway. To investigate the involvement of these proteins in DNA fragmentation after ischemia/reperfusion, Ku protein expression was examined before and after transient focal cerebral ischemia (FCI) in mice. METHODS: Adult male CD-1 mice were subjected to 60 minutes of FCI by intraluminal suture blockade of the middle cerebral artery. Ku protein expression was studied by immunohistochemistry and Western blot analysis. DNA fragmentation was evaluated by gel electrophoresis and terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). The spatial relationship between Ku expression and DNA fragmentation was examined by double labeling with Ku and TUNEL after reperfusion. RESULTS: Immunohistochemistry showed constitutive expression of Ku proteins in control brains. The number of Ku-expressing cells was decreased in the entire middle cerebral artery territory as early as 4 hours after reperfusion and remained reduced until 24 hours. Western blot analyses confirmed the significant reduction of these proteins (59.4% and 57.7% reduction in optical density at 4 hours of reperfusion from the normal level of Ku70 and Ku86 bands, respectively; P<0.001). DNA gel electrophoresis demonstrated DNA laddering 24 hours after reperfusion, but not at 4 hours. Double staining with Ku and TUNEL showed a concomitant loss of Ku immunoreactivity and TUNEL-positive staining. CONCLUSIONS: These results suggest that the early reduction of Ku proteins and the loss of defense against DNA damage may underlie the mechanism of DNA fragmentation after FCI. PMID- 11387506 TI - Vasopressin-induced protein kinase C-dependent superoxide generation contributes to atp-sensitive potassium channel but not calcium-sensitive potassium channel function impairment after brain injury. AB - BACKGROUND AND PURPOSE: Pial artery dilation in response to activators of the ATP sensitive K(+) (K(ATP)) and calcium-sensitive K(+) (K(Ca)) channels is impaired after fluid percussion brain injury (FPI). Vasopressin, when coadministered with the K(ATP) and K(Ca) channel agonists cromakalim and NS1619 in a concentration approximating that observed in cerebrospinal fluid (CSF) after FPI, blunted K(ATP) and K(Ca) channel-mediated vasodilation. Vasopressin also contributes to impaired K(ATP) and K(Ca) channel vasodilation after FPI. In addition, protein kinase C (PKC) activation generates superoxide anion (O(2)(-)), which in turn contributes to K(ATP) channel impairment after FPI. We tested whether vasopressin generates O(2)(-) in a protein kinase C (PKC)-dependent manner, which could link vasopressin release to impaired K(ATP) and K(Ca) channel-induced pial artery dilation after FPI. METHODS: Injury of moderate severity (1.9 to 2.1 atm) was produced with the lateral FPI technique in anesthetized newborn pigs equipped with a closed cranial window. Superoxide dismutase-inhibitable nitroblue tetrazolium (NBT) reduction was determined as an index of O(2)(-) generation. RESULTS: Under sham injury conditions, topical vasopressin (40 pg/mL, the concentration present in CSF after FPI) increased superoxide dismutase inhibitable NBT reduction from 1+/-1 to 23+/-4 pmol/mm(2). Chelerythrine (10(-7) mol/L, a PKC inhibitor) blunted such NBT reduction (1+/-1 to 9+/-2 pmol/mm(2)), whereas the vasopressin antagonist l-(beta-mercapto-beta,beta-cyclopentamethylene propionic acid)2-(o-methyl)-Tyr-arginine vasopressin (MEAVP) blocked NBT reduction. Chelerythrine and MEAVP also blunted the NBT reduction observed after FPI (1+/-1 to 15+/-1, 1+/-1 to 4+/-1, and 1+/-1 to 5+/-1 pmol/mm(2) for sham-, chelerythrine-, and MEAVP-treated animals, respectively). Under sham injury conditions, vasopressin (40 pg/mL) coadministered with cromakalim or NS1619 blunted dilation in response to these K(+) channel agonists, whereas chelerythrine partially restored such impaired vasodilation for cromakalim but not NS1619. Cromakalim- and NS1619-induced pial artery dilation also was blunted after FPI. MEAVP partially protected dilation to both K(+) channel agonists after FPI, whereas chelerythrine did so for only cromakalim responses (for cromakalim at 10(-8) and 10(-6) mol/L, 13+/-1% and 23+/-1%, 2+/-1% and 5+/-1%, 9+/-1% and 15+/-2%, and 9+/-1% and 16+/-2% for sham-, FPI-, FPI-MEAVP-, and FPI chelerythrine-pretreated animals, respectively). CONCLUSIONS: These data show that vasopressin, in concentrations present in CSF after FPI, increased O(2)(-) production in a PKC-dependent manner and contributes to such production after FPI. These data show that vasopressin contributes to K(ATP) but not K(Ca) channel function impairment in a PKC-dependent manner after FPI and suggest that vasopressin contributes to K(Ca) channel function impairment after FPI via a mechanism independent of PKC activation. PMID- 11387507 TI - Towards a national system for monitoring the quality of hospital-based stroke services. AB - BACKGROUND AND PURPOSE: We sought to evaluate a system for monitoring the quality of hospital-based stroke services that uses routinely collected case fatality data, adjusted for case mix, as well as simple measures of the process of stroke care. METHODS: We compared the process of care and case fatality after stroke between 5 Scottish hospitals (A through E) during 1995-1997. We retrospectively identified 2724 patients with acute stroke using routine discharge data and obtained case mix and process of care data from the medical record. We ascertained case fatality by record linkage and adjusted for case mix using a simple, externally validated regression model. RESULTS: Crude case fatality varied by 21 deaths per 100 admissions between the 5 hospitals. After adjustment, case fatality still differed significantly (P=0.047), with 5 to 7 more deaths per 100 admissions at Hospital A than at Hospitals B through E. There were major shortcomings in the specialization and organization of care, the use of CT scanning, and the completeness of documentation at Hospital A compared with the other hospitals. There were smaller, but clinically important, differences in care between Hospitals B through E but no significant differences in adjusted case fatality. CONCLUSIONS: Once adjusted for important prognostic variables, routinely collected case fatality data might identify hospitals with major shortcomings in the processes of stroke care. More moderate, but still clinically important, variations in stroke care can only be identified by monitoring the process of care directly. PMID- 11387508 TI - Clinically unidentified dissection of vertebral artery as a cause of cerebellar infarction. AB - BACKGROUND AND PURPOSE: Dissection of vertebral arteries has been reported in association with minor neck movements without signs of trauma on the surface of the neck. In addition, injury of a vertebral artery can cause brain infarctions. However, few cases have been reported in which fatal brain infarction was due to nonocclusive, clinically undetected, traumatic thrombus formation in a vertebral artery. CASE DESCRIPTION: A 62-year-old man was hit by a car, and a right cerebellar infarction was found the day after the accident. The cause of the infarction could not be detected by angiography. Although the patient recovered favorably after surgical removal of the right lateral hemisphere of the cerebellum, he died suddenly 2 weeks after the accident. An autopsy and a microscopic study revealed pulmonary thromboembolism and organizing traumatic lesions of the right vertebral artery without occlusion or noteworthy stenosis of the artery. CONCLUSIONS: We concluded that the patient sustained traumatic lesions of the right vertebral artery during the traffic accident 2 weeks before death and that his cerebellar infarction was due to a thrombus resulting from these traumatic lesions. PMID- 11387509 TI - The utility of health states after stroke: a systematic review of the literature. AB - BACKGROUND: To perform decision analyses that include stroke as one of the possible health states, the utilities of stroke states must be determined. We reviewed the literature to obtain estimates of the utility of stroke and explored the impact of the study population and the elicitation method. SUMMARY OF REVIEW: We searched various databases for articles reporting empirical assessment of utilities. Mean utilities of major stroke (Rankin Scale 4 to 5) and minor stroke (Rankin Scale 2 to 3) were calculated, stratified by study population and elicitation method. Additionally, the modified Rankin Scale was mapped onto the EuroQol classification system. Utilities were obtained from 23 articles. Patients at risk for stroke assigned utilities of 0.26 and 0.55 to major and minor stroke, respectively. Stroke survivors assigned higher utilities to both major (0.41) and minor stroke (0.72). The EuroQol completed by stroke survivors revealed a utility of 0.32 and 0.71 for major and minor stroke, respectively. Utilities elicited by the Standard Gamble were generally higher, while those obtained by the Visual Analogue Scale were lower than the Time Trade Off values. Remaining variation between utilities may be caused by differences in definitions of the health states. The mapped EuroQol indicated a utility of 0.64 for minor stroke and a value just below zero for major stroke. CONCLUSIONS: For minor stroke, a utility between 0.50 and 0.70 seems to be reasonable for both decision analyses and cost effectiveness studies. The utility of major stroke may range between 0 and 0.30 and may possibly be negative. PMID- 11387511 TI - Systemic complement activation in ischemic stroke. PMID- 11387510 TI - Reporting terminology for brain arteriovenous malformation clinical and radiographic features for use in clinical trials. PMID- 11387512 TI - Venous thromboembolism after acute stroke. PMID- 11387513 TI - Re: cost and outcome of mechanical ventilation for life-threatening stroke. PMID- 11387514 TI - Effect of contralateral occlusion on long-term efficacy of endarterectomy in the Asymptomatic Carotid Atherosclerosis Study (ACAS). PMID- 11387515 TI - Diurnal variation in stroke onset in atrial fibrillation. PMID- 11387518 TI - Clearing the smoke or muddying the water? PMID- 11387517 TI - AHA Scientific Statement: Recommendations for the management of intracranial arteriovenous malformations: a statement for healthcare professionals from a special writing group of the Stroke Council, American Stroke Association. PMID- 11387519 TI - Hard targets. PMID- 11387528 TI - Whose standard is it, anyway? How the tobacco industry determines the International Organization for Standardization (ISO) standards for tobacco and tobacco products. AB - OBJECTIVE: To describe the extent of the tobacco industry involvement in establishing international standards for tobacco and tobacco products and the industry influence on the International Organization for Standardization (ISO). METHODS: Analysis of tobacco industry documents made public as part of the settlement of the Minnesota Tobacco Trial and the Master Settlement Agreement. Search words included "ISO", "CORESTA", "Barclay", "compensation and machine smoking", "tar and nicotine deliveries", and the name of key players, in different combinations. RESULTS: It is clear that the tobacco industry, through the Cooperation Centre for Scientific Research Relative to Tobacco (CORESTA), play a major role in determining the scientific evidence and suggesting the standards that are eventually adopted as international standards for tobacco and tobacco products in several areas, including the measurement of cigarette tar and nicotine yield. CONCLUSIONS: ISO's tobacco and tobacco products standards are not adequate to guide tobacco products regulatory policies, and no health claims can be made based on ISO's tobacco products standards. There is an urgent need for tobacco control advocates and groups worldwide to be more involved with the work of the ISO, both directly and through their national standardisation organisations. PMID- 11387529 TI - Challenging public health acceptability of current international standards on tobacco products: paving the way for strengthened cooperation. PMID- 11387530 TI - Effect of policies directed at youth access to smoking: results from the SimSmoke computer simulation model. AB - OBJECTIVES: To develop a simulation model to predict the effects of youth access policies on retail compliance, smoking rates, and smoking attributable deaths. METHODS: A model of youth access policies is developed based on empirical research and a theory of perceived risk. The model incorporates substitution into other sources as retail sales are restricted, and is used to project the number of smokers and smoking related deaths. Various policies to limit youth access to cigarettes are evaluated, and we explore how efficient policies may be developed. RESULTS: The model predicts that a well designed and comprehensive policy that includes sufficient compliance checks, penalties, and community involvement has the potential to reduce the number of young smokers. Because smoking related deaths occur later in life, the effects on health are largely delayed. CONCLUSIONS: A well designed youth access policy has the ability to affect youth smoking rates in the short term, and will lead to savings in lives in future years. The ability of retail oriented policies to reduce youth smoking, however, is limited. Other tobacco control policies, including those directed at non retail sources of cigarettes, are also needed. PMID- 11387531 TI - School smoking policies and smoking prevalence among adolescents: multilevel analysis of cross-sectional data from Wales. AB - OBJECTIVE: To examine the association between school smoking policies and smoking prevalence among pupils. DESIGN: Multilevel analysis of cross-sectional data from surveys of schools and pupils. SETTING: 55 secondary schools in Wales. SUBJECTS: 55 teachers and 1375 pupils in year 11 (aged 15-16). MAIN OUTCOME MEASURES: Self reported smoking behaviour. RESULTS: The prevalence of daily smoking in schools with a written policy on smoking for pupils, teachers, and other adults, with no pupils or teachers allowed to smoke anywhere on the school premises, was 9.5% (95% confidence interval (CI) 6.1% to 12.9%). In schools with no policy on pupils' or teachers' smoking, 30.1% (95% CI 23.6% to 36.6%) of pupils reported daily smoking. In schools with an intermediate level of smoking policy, 21.0% (95% CI 17.8% to 24.2%) smoked every day. School smoking policy was associated with school level variation in daily smoking (p = 0.002). In multilevel analysis, after adjusting for pupils' sex, parents' and best friends' smoking status, parental expectations, and alienation from school, there was less unexplained school level variation, but school smoking policy remained significant (p = 0.041). The association of smoking policy with weekly smoking was weaker than for daily smoking, and not significant after adjustment for pupil level variables. Both daily and weekly smoking prevalence were lower in schools where pupils' smoking restrictions were always enforced. Enforcement of teacher smoking restrictions was not significantly associated with pupils' smoking. CONCLUSIONS: This study demonstrates an association between policy strength, policy enforcement, and the prevalence of smoking among pupils, after having adjusted for pupil level characteristics. These findings suggest that the wider introduction of comprehensive school smoking policies may help reduce teenage smoking. PMID- 11387532 TI - Tobacco lobby political influence on US state legislatures in the 1990s. AB - BACKGROUND: Throughout the 1990s the tobacco lobby was a potent political force in US state legislatures advancing its pro-tobacco agenda. OBJECTIVE: To describe the market and political motivations of the tobacco lobby and the strategies they use to achieve these goals in US state legislatures. DESIGN: This study is a content analysis and summary overview of recently released historical tobacco industry documents; tobacco related government documents; and recent state tobacco control policy reports. RESULTS: In the 1990s, the tobacco lobby engaged in a comprehensive and aggressive political effort in state legislatures to sell tobacco with the least hindrance using lobbying, the media, public relations, front groups, industry allies, and contributions to legislators. These efforts included campaigns to neutralise clean indoor air legislation, minimise tax increases, and preserve the industry's freedom to advertise and sell tobacco. The tobacco lobby succeeded in increasing the number of states that enacted state pre emption of stricter local tobacco control laws and prevented the passage of many state tobacco control policies. Public health advocates were able to prevent pre emption and other pro-tobacco policies from being enacted in several states. CONCLUSIONS: The tobacco lobby is a powerful presence in state legislatures. Because of the poor public image of the tobacco lobby, it seeks to wield this power quietly and behind the scenes. State and local health advocates, who often have high public credibility, can use this fact against the tobacco lobby by focusing public attention on the tobacco lobby's political influence and policy goals and expose links between the tobacco lobby and its legislative supporters. PMID- 11387533 TI - The price of cigarettes in the European Union. AB - BACKGROUND: A major factor influencing tobacco use is its price. Fiscal policies on tobacco are a key ingredient of any comprehensive control strategy, as they can be used to raise prices. The European Union (EU) developed directives to ensure some harmonisation of the fiscal pressure on tobacco across its member states. OBJECTIVES: To provide a simple comparison of tobacco prices in the EU, adjusting for the purchasing power of each currency. DESIGN: For price comparisons, a 20 units pack of Marlboro was the reference product, and data refer to April 2000. Purchasing power parities (PPP) for each member state currency have been compiled. These are currency conversion rates, which convert to a common currency and equalise the purchasing power of different currencies. MAIN OUTCOME MEASURES: Nominal prices of a Marlboro pack for each member state, and a price index, estimated taking as reference the EU mean. Adjusted prices and an adjusted price index have been estimated using PPP. RESULTS: Nominal prices show wide variation, with the cheapest pack in Portugal (59) and the most expensive in the UK (196); the range of variation is three-fold. However, PPP adjusted prices reveal a different distribution. In three countries adjusted prices are outliers, but all other countries make two clusters, one around the average EU index of 100, the other around a lower value of 85. CONCLUSIONS: These results suggest that fiscal harmonisation policies in the EU do not have an even effect at reducing availability by its impact in price. PMID- 11387534 TI - Can media advocacy influence newspaper coverage of tobacco: measuring the effectiveness of the American stop smoking intervention study's (ASSIST) media advocacy strategies. AB - OBJECTIVE: To compare the rate and slant of local tobacco control print media coverage in ASSIST (American stop smoking intervention study) states as compared with non-ASSIST states. METHODS: Local tobacco control policy articles, editorials, and letters to the editors published from 1994 to 1998 clipped from all daily local newspapers in the USA were analysed (n = 95 911). The main hypothesis tested for the existence of an interaction between ASSIST intervention and time. This interaction would represent a change in the difference between ASSIST and non-ASSIST states over the course of the intervention. RESULTS: No evidence of an ASSIST-year interaction was found. However, a main effect for ASSIST was significant for the percentage of articles with the model predicting higher rates of articles for ASSIST states. Similarly the rate of letters to the editor expressing protobacco control views was higher in ASSIST states than non ASSIST states. No main effects or interactions were found for analyses of percentage of protobacco control editorials. Models controlled for a measure of preintervention tobacco control conditions at baseline. CONCLUSIONS: The presence of an ASSIST main effect should be interpreted with caution because of the quasi experimental design and the lack of information on article rates before the ASSIST intervention. Nonetheless, these preliminary findings suggest some possible effects of the media advocacy activities of ASSIST when controlling for differences in states' initial tobacco control conditions. PMID- 11387535 TI - News media coverage of smoking and health is associated with changes in population rates of smoking cessation but not initiation. AB - OBJECTIVE: To determine whether changes in news media coverage of smoking and health issues are associated with changes in smoking behaviour in the USA. DESIGN AND MAIN OUTCOME MEASURES: Issue importance in the US news media is assessed by the number of articles published annually in major magazines indexed in The Reader's Guide to Periodical Literature. Annual incidence rates for cessation and initiation in the USA were computed from the large, representative National Health Interview Surveys (1965-1992). Patterns in cessation incidence were considered for ages 20-34 years and 35-50 years. Initiation incidence was examined for adolescents (14-17 years) and young adults (18-21 years) of both sexes. RESULTS: From 1950 to the early 1980s, the annual incidence of cessation in the USA mirrored the pattern of news media coverage of smoking and health, particularly for middle aged smokers. Cessation rates in younger adults increased considerably when second hand smoke concerns started to increase in the US population. Incidence of initiation in young adults did not start to decline until the beginning of the public health campaign against smoking in the 1960s. Among adolescents, incidence rates did not start to decline until the 1970s, after the broadcast ban on cigarette advertising. CONCLUSIONS: The level of coverage of smoking and health in the news media may play an important role in determining the rate of population smoking cessation, but not initiation. In countries where cessation has lagged, advocates should work to increase the newsworthiness of smoking and health issues. PMID- 11387536 TI - Print media coverage of California's smokefree bar law. AB - OBJECTIVE: To assess the print media coverage of California's smokefree bar law in the state of California. DESIGN: Content analysis of newspaper, trade journal, and magazine items. SUBJECTS: Items regarding the smokefree bar law published seven months before and one year following the implementation of the smokefree bar law (June 1997 to December 1998). Items consisted of news articles (n = 446), opinion editorials (n = 31), editorials (n = 104), letters to the editor (n = 240), and cartoons (n = 10). MAIN OUTCOME MEASURES: Number and timing of publication of items, presence of tobacco industry arguments or public health arguments regarding law, positive, negative, and neutral views of opinion items published. RESULTS: 53% of items published concerning the smokefree bar law were news articles, 47% were opinion items. 45% of items regarding the smokefree bar law were published during the first month of implementation. The tobacco industry dominated coverage in most categories (economics, choice, enforcement, ventilation, legislation, individual quotes), except for categories public health used the most frequently (government role, tactics, organisational quotes). Anti law editorials and letters to the editor were published more than pro-law editorials and letters. Region of the state, paper size, presence of local clean indoor air legislation, and voting on tobacco related ballot initiatives did not have an impact on the presence of opinion items. CONCLUSIONS: The tobacco industry succeeded in obtaining more coverage of the smokefree bar law, both in news items and opinion items. The tobacco industry used historical arguments of restricting freedom of choice and economic ramifications in fighting the smokefree bar law, while public health groups focused on the worker protection issue, and exposed tobacco industry tactics. Despite the skewed coverage, public health groups obtained adequate attention to their arguments to keep the law in effect. PMID- 11387537 TI - Comparison of physician based reporting of tobacco attributable deaths and computer derived estimates of smoking attributable deaths, Oregon, 1989 to 1996. AB - BACKGROUND: Tobacco use prevention programmes need accurate information about smoking related mortality. Beginning in 1989, Oregon began asking physicians to report on death certificates whether tobacco use contributed to the death. OBJECTIVE: To determine the long term comparability of this method of estimating tobacco attributable mortality to estimates of smoking attributable mortality derived from a computer model. DESIGN: For the period 1989 to 1996, we compared mortality resulting from tobacco use reported by Oregon physicians to estimates of smoking attributable deaths (SADs) derived by "Smoking attributable mortality, morbidity and economic costs" software version 3.0 (SAMMEC 3.0), a widely used software program that estimates SADs on the basis of smoking prevalence and relative risks of specific diseases among current and former smokers. MAIN OUTCOME MEASURES: Numbers of deaths, age, sex, and category of disease. RESULTS: Of 212, 448 Oregon deaths during 1989-1996, SAMMEC 3.0 estimated that 42, 778 (20.1%) were attributable to cigarette smoking. For the same 27 diagnoses, physicians reported that tobacco contributed to 42, 839 (20.2%) deaths-a cumulative difference of only 61 deaths over the eight year period. The age and sex distributions of tobacco and smoking attributable deaths reported by the two systems were also similar. By category of disease, the ratio of SAMMEC 3.0 estimates to physician reported deaths was 1.11 for neoplasms, 0.88 for heart disease, and 1.04 for respiratory disease. CONCLUSIONS: Physician reporting provides comparable estimates of smoking attributable mortality and can be a valuable source of data for communicating the risks of tobacco use to the public. PMID- 11387538 TI - Sociodemographic predictors of success in smoking intervention. AB - AIM: To examine the role of sociodemographic factors as predictors of sustained smoking cessation for the population who volunteer to participate in intervention programmes. METHOD: Data for the 3575 smokers who participated in the CEASE (collaborative European anti-smoking evaluation) trial, a European multicentred study that used transdermal nicotine patches as an adjunct to smoking cessation in the chest clinic, were analysed. The effects of age, sex, smoking habit, socioeconomic status (housing conditions, education, and employment), disease, smoking habits of relatives, and baseline markers of tobacco use on sustained smoking cessation (self-reported abstinence and expired carbon monoxide < 10 parts per million) were assessed using logistic regression modelling (odds ratio (OR), 95% confidence interval (CI)). RESULTS: 477/3575 smokers were sustained abstainers one year after the intervention (overall success rate 13.3%). In the univariable logistic regression models an effect of active treatment on smoking cessation was observed (OR 1.50, 95% CI 1.15 to 1.96), and additional effects on outcome were found for age (OR 1.02, 95% CI 1.01 to 1.03), sex (men v women: OR 1.38, 95% CI 1.14 to 1.68), housing conditions (OR 1.43, 95% CI 1.25 to 1.65), current respiratory (OR 0.79, 95% CI 0.67 to 0.92) or cardiac disease (OR 0.46, 95% CI 0.28 to 0.75), and markers of tobacco use (cigarettes per day: OR 0.79, 95% CI 0.69 to 0.90; expired carbon monoxide: OR 0.98, 95% CI 0.97 to 0.99). Education and employment did not have a significant effect on the outcome. The effect of the variables associated with success in smoking cessation persisted after adjustment for covariates. CONCLUSION: Age, sex, and housing conditions have a major effect on smoking cessation in European smokers participating in smoking cessation programmes. PMID- 11387539 TI - Smoking cessation in China: findings from the 1996 national prevalence survey. AB - OBJECTIVES: To describe patterns of smoking and smoking cessation in China within the context of the stages of change model, using data from the 1996 national prevalence survey. DESIGN: A cross sectional survey was carried out using the 145 preselected disease surveillance points, which provide a representative sample for the entire country. A standardised questionnaire on smoking was interviewer administered. SETTING: The country of China. SUBJECTS: 122,220 people aged 15-69 years. MAINTENANCE MEASURES: Smoking cessation patterns, as defined by smoking status (current or former) and stage of change (precontemplation, contemplation, and action). RESULTS: The sample included 45,995 ever smokers of whom 4336 had quit. About 72% of current smokers reported not intending to give up their smoking behaviour, and about 16% of current smokers said they intended to do so, but have not taken any action. Of all ever smokers, the percentage of former smokers was 9.5%, and 12% of current smokers had quit at least once, but relapsed by the time of the survey. The patterns were similar in men and women with regard to the stated intent to quit. Among males, the percentage of former smokers increased with age but the percentage intending to quit was constant at about 15% across age strata. The most common reason for quitting was illness. Participants with a university education were more likely to have made an attempt to quit. CONCLUSIONS: The percentage of smokers contemplating quitting was low in China in 1996. The study shows that smokers in China must be mobilised to contemplate quitting and then to take action. PMID- 11387540 TI - Variations in treatment benefits influence smoking cessation: results of a randomised controlled trial. AB - OBJECTIVE: To assess the impact and costs of coverage for tobacco dependence treatment benefits with no patient cost sharing for smokers with employer sponsored coverage in two large independent practice association (IPA) model health maintenance organisations (HMOs) in California, USA. METHODS: A randomised experimental design was used. 1204 eligible smokers were randomly assigned either to the control group, which received a self-help kit (video and pamphlet), or to the treatment group, which received the self-help kit and fully covered benefits for over the counter (OTC) nicotine replacement therapy (NRT) gum and patch, and participation in a group behavioural cessation programme with no patient cost sharing. RESULTS: The quit rates after one year of follow up were 18% in the treatment group and 13% in the control group (adjusted odd ratio (OR) 1.6, 95% confidence interval (CI) 1.1 to 2.4), controlling for health plan, sociodemographics, baseline smoking characteristics, and use of bupropion. Rates of quit attempts (adjusted OR 1.4, 95% CI 1.1 to 1.8) and use of nicotine gum or patch (adjusted OR 2.3, 95% CI 1.6 to 3.2) were also higher in the treatment group. The annual cost of the benefit per user who quit ranged from $1495 to $965 or from $0.73 to $0.47 per HMO member per month. CONCLUSIONS: Full coverage of a tobacco dependence treatment benefit implemented in two IPA model HMOs in California has been shown to be an effective and relatively low cost strategy for significantly increasing quit rates, quit attempts, and use of nicotine gum and patch in adult smokers. PMID- 11387541 TI - Comparison of the nicotine content of tobacco used in bidis and conventional cigarettes. AB - OBJECTIVE: To compare the nicotine content of 12 unfiltered brands of bidi cigarettes (hand rolled cigarettes imported from India) with 8 popular brands of filtered and unfiltered US and conventional cigarettes from India. MAIN OUTCOME MEASURES: Identical laboratory procedures were used to determine nicotine content (in duplicate) and physical characteristics. RESULTS: The nicotine concentration in the tobacco of bidi cigarettes (21.2 mg/g) was significantly greater than the tobacco from the commercial filtered (16.3 mg/g) and unfiltered cigarettes (13.5 mg/g). CONCLUSIONS: Bidi cigarettes contain higher concentrations of nicotine than conventional cigarettes. Therefore, it is logical to presume that bidi smokers are at risk of becoming nicotine dependent. These findings belief a popular belief among US teens that bidis are a safe alternative to commercial cigarettes. PMID- 11387542 TI - Cigarette advertising and promotional strategies in retail outlets: results of a statewide survey in California. AB - OBJECTIVE: To examine the extent and types of cigarette advertising materials in stores and to assess tobacco company compliance with the 1998 Master Settlement Agreement (MSA). DESIGN: A cross-sectional analysis of a random sample of 586 stores that sold cigarettes. SETTING: US state of California. MAIN OUTCOME MEASURES: Trained data collectors classified cigarette advertising materials by type (signs, displays, functional items), location (interior or exterior), and placement (below 3 feet (1 m) or near candy). RESULTS: California retail outlets featured 17.2 (SD 16.1) tobacco advertising materials on average, and 94% of stores featured at least some advertising. About 85% of these were within 4 feet (1.3 m) of the counter. About 50% of the stores had ads at or below 3 feet, and 23% had cigarette product displays next to candy. In violation of the MSA, 3% of stores featured signs with cartoons and 11% had large exterior signs. CONCLUSIONS: Tobacco companies are aggressively using stores to market cigarettes. Moreover, the spirit of the MSA-to protect children from cigarette advertising-has not been realised. Future studies should monitor industry use of this venue and assess the impact of exposure to cigarette advertising materials in stores on adult smokers and youth. PMID- 11387543 TI - Clearing the smoke: the science base for tobacco harm reduction--executive summary. PMID- 11387544 TI - Marketing life after advertising bans. PMID- 11387545 TI - Choroid plexus ependymal cells enhance neurite outgrowth from dorsal root ganglion neurons in vitro. AB - The epithelial cells of the choroid plexus are a continuation of the ventricular ependymal cells and are regarded as modified ependymal cells. The present study was carried out to determine the influence of choroid plexus ependymal cells (CPECs) on axonal growth in vitro. Choroid plexuses were dissected from the fourth ventricle of postnatal day-1-10 mice, mechanically dissociated, and plated in fibronectin-coated culture dishes. CPECs had spread into monolayers with few endothelial cells in 3-week cultures. Some macrophages were scattered on the monolayer of CPECs. Dorsal root ganglia (DRG) were excised from mouse fetuses of 14-day gestation, dissociated with trypsin and cocultured on the CPEC monolayers. For comparison, dissociated DRG neurons were cocultured on astrocyte monolayers or cultured on laminin-coated plates. After 4.5 h culturing, the cultures were fixed and immunohistochemically double-stained for neurites and CPECs using antibodies against beta-tubulin III and S-100 beta, respectively. It was demonstrated that neurons extended many long neurites with elaborate branching on the surface of S-100-stained CPECs. In contrast, DRG neurons cultured on the astrocytes and on the laminin-coated plates had much shorter primary neurites with fewer branches than those cultured on the CPECs. The total length of neurites including primary neurites and their branches, of a single DRG neuron was 285 +/- 14, 395 +/- 15 and 565 +/- 12 microM on the laminin-coated plates, on astrocytes and on CPECs, respectively. Scanning electron microscopy revealed extension of neurites with well-developed growth cones on the ependymal cells. These results suggest that CPECs have a great capacity to promote neurite outgrowth from DRG neurons in vitro. PMID- 11387546 TI - Alpha 1E subunit of the R-type calcium channel is associated with myelinogenesis. AB - During myelinogenesis, we found an exceedingly strong, transient expression of the alpha(1E) gene for the R-type voltage-gated calcium channel in CNS white matter. This immunoreactivity appeared in glial cells along specific pathways of the brainstem, cerebellum, and telencephalon. The reactivity followed a wave that progressed from the brainstem at P5, to the cerebellar peduncles by P8, the arbor vitae by P14, and the granular layer by P17. The reactivity-peaked about 3-4 days later and decreased gradually to become negligible in all areas before adulthood. Ultrastructural analysis confirmed that alpha(1E) immunoreactivity was located in oligodendroglial somata, their projections, paranodal wraps and loose myelin sheaths. There was a distinct association of the channel protein reactivity on oligodendroglial membranes in contact with the axon. We propose that glial projections, contacting axons, sense axonal firing through small K(+) currents and open the high voltage R-type calcium channels to signal myelination. PMID- 11387548 TI - A light and electron microscopic study of glutamate receptors in the monkey subthalamic nucleus. AB - The distribution of glutamate receptors in the monkey subthalamic nucleus was studied using affinity purified polyclonal antibodies to GluR1, phosphorylated GluR1, GluR2/3, NMDAR1, mGluR1a and mGluR5. Intense staining for both the unphosphorylated and the phosphorylated forms of the AMPA receptor subunit GluR1 was observed in the cell bodies and proximal dendrites of neurons in this nucleus. In comparison to GluR1, less intense staining for GluR2/3 was observed in the cell bodies and processes. NMDAR1 immunoreactivity was present in cell bodies and large numbers of small diameter dendrites. Light staining was observed in cell bodies with mGluR1a and no staining was observed on cell bodies with mGluR5. The neuropil, however, contained many processes that were labeled for mGluR1a or mGluR5. Electron microscopy showed that label was present in cytoplasmic locations in cell bodies and dendrites, in addition to components of the synaptic region, in sections stained for GluR1, GluR2/3 and NMDAR1. In contrast, very lightly labeled or unlabeled cell bodies but labeled dendrites and axon terminals, was observed in sections stained for mGluR1a and mGluR5. In addition to neural processes, occasional astrocytic processes were also labeled for mGluR5. Of the immunogold particles that were associated with components of the synaptic region, label for ionotropic glutamate receptors was mostly present on postsynaptic densities, whilst that for metabotropic glutamate receptors was mostly present in a perisynaptic location. The ratio of GluR1/GluR2 messenger RNAs has been reported to increase in the aged hippocampus (PAGLIUSI, S. R., GERRARD, P., ABDALLAH, M., TALABOT, D. & CATSICAS, S. (1994) Neuroscience 61, 429 433.), and it is possible that a similar change in the ratio of GluR1 and GluR2 may occur in neurons of the subthalamic nucleus with age. It is postulated that this could result an increase in calcium permeability via AMPA receptors, and an enhancement of excitatory transmission in this nucleus. PMID- 11387547 TI - Expression and localization of ionotropic glutamate receptor subunits in the goldfish retina--an in situ hybridization and immunocytochemical study. AB - The expression and distribution of AMPA, kainate and NMDA glutamate receptor subunits was studied in the goldfish retina. For the immunocytochemical localization of the AMPA receptor antisera against GluR2, GluR2/3 and GluR4 were used, and for in situ hybridization rat specific probes for GluR1 and GluR2 and goldfish specific probes for GluR3 and GluR4 were used. The localization of the low affinity kainate receptor and NMDA receptor was studied using antisera against GluR5-7 and NR1. All AMPA receptor subtypes were demonstrated to be present in the goldfish retina both by immunocytochemistry and in situ hybridization. In situ hybridization revealed expression of all AMPA receptors subunit at the inner border of the INL. Only GluR3 was also strongly expressed in the outer border of the INL. Some of the ganglion cells displayed a strong signal for GluR1, GluR3 and GluR4. GluR1-immunoreactivity was present in subsets of bipolar, amacrine, and ganglion cells. GluR2 and GluR2/3-immunoreactivity was mainly localized in the outer plexiform layer. GluR2 and GluR2/3-immunoreactivity are associated with the photoreceptor synaptic terminals. GluR4-immunoreactivity is present on Muller cells in the inner retina and on dendrites of bipolar cells in the OPL, whereas GluR5-7-immunoreactivity was prominently present on horizontal cell axon terminals. Finally, NR1-immunoreactivity was confined to amacrine cells, the inner plexiform layer and ganglion cells. This study shows that there is a strong heterogeneity of glutamate receptor subunit expression in the various layers of the retina. Of the AMPA receptor subunits GluR3 seems to be expressed the most widely in all layers with strong glutamatergic synaptic interactions whereas all the other subunits seem to have a more restricted expressed pattern. PMID- 11387549 TI - Myelination of prospective large fibres in chicken ventral funiculus. AB - In mammals, the oligodendrocyte population includes morphological and molecular varieties. We reported previously that an antiserum against the T4-O molecule labels a subgroup of oligodendrocytes related to large myelinated axons in adult chicken white matter. We also reported that T4-O immunoreactive cells first appear in the developing ventral funiculus (VF) at embryonic day (E)15, subsequently increasing rapidly in number. Relevant fine structural data for comparison are not available in the literature. This prompted the present morphological analysis of developing and mature VF white matter in the chicken. The first axon-oligodendrocyte connections were seen at E10 and formation of compact myelin had started at E12. Between E12 and E15 the first myelinating oligodendrocytes attained a Schwann cell-like morphology. At hatching (E21) 60% of all VF axons were myelinated and in the adult this proportion had increased to 85%. The semilunar or polygonal oligodendrocytes associated with adult myelinated axons contained many organelles indicating a vivid metabolic activity. Domeshaped outbulgings with gap junction-like connections to astrocytic profiles were frequent. Oligodendrocytes surrounded by large myelinated axons and those surrounded by small myelinated axons were cytologically similar. But, thick and thin myelin sheaths had dissimilar periodicities and Marchi-positive myelinoid bodies occurred preferentially in relation to large myelinated axons. We conclude that early oligodendrocytes contact axons and form myelin well before the first expression of T4-O and that emergence of a T4-O immunoreactivity coincides in time with development of a Type IV phenotype. Our data also show that oligodendrocytes associated with thick axons are cytologically similar to cells related to thin axons. In addition, the development of chicken VF white matter was found to be similar to the development of mammalian white matter, except for the rapid time course. PMID- 11387550 TI - Regional distribution of NPC1 protein in monkey brain. AB - NPC1 is a member of a family of polytopic membrane-bound proteins with sterol sensing domains. Inactivating mutations of NPC1 are responsible for most cases of Niemann-Pick type C disease, whose hallmark is progressive neurodegeneration. The precise molecular mechanisms whereby defective NPC1 function leads to neurodegeneration are unknown. In the brain, we have previously found NPC1 to localize predominantly within perisynaptic astrocytic processes. Here we have mapped the regional distribution of NPC1 in the monkey brain. Dense NPC1 immunoreactivity was observed in telencephalic structures, including the cerebral neocortex, hippocampus, caudate nucleus and putamen, whilst light immunostaining was observed in diencephalic structures, including the globus pallidus, thalamus and hypothalamus. Light staining was also generally observed in the midbrain, pons, medulla oblongata and cerebellum, except the inferior olive, which was densely stained. By light microscopy, only a few indistinctly labeled cell bodies were observed even within densely labeled regions, where most of the immunoreactivity appeared to be due to the large numbers of labeled cellular processes. On electron microscopy, these processes were identified as glial, and not neuronal. The astrocytic localization of NPC1 was further confirmed by double labeling for NPC1 and GFAP. The regional pattern of NPC1 expression suggests that areas normally expressing low levels of the NPC1 protein are more susceptible to neuronal degeneration in Niemann-Pick type C disease. PMID- 11387551 TI - Nursing's current state of affairs. PMID- 11387552 TI - Overcrowding and other crises: how can we survive? PMID- 11387553 TI - Research data suggest prehospital endotracheal intubation for children no more effective than bag-valve-mask. PMID- 11387555 TI - More on children's bullying. PMID- 11387556 TI - Breast cancer study seeks volunteers for short survey. PMID- 11387557 TI - More on children's bullying. PMID- 11387558 TI - An ED nurse reflects on the murders of an infant and a toddler. PMID- 11387559 TI - A 22-year-old woman with exquisite burning pain 4 weeks after an ankle sprain. PMID- 11387560 TI - Assessment of family-centered care policies and practices for pediatric patients in nine US emergency departments. AB - INTRODUCTION: Family-centered care recognizes the integral role of the family in the health and well-being of the pediatric patient. However, implementing a family-centered care approach often requires significant philosophical, practice, and environmental changes. A self-assessment inventory of family-centered practices can identify areas for change. METHODS: ENA conducted a validation study on an instrument used to assess family-centered care in the emergency department. Nine emergency departments of varying demographics used the Family centered Care Self-assessment Inventory to evaluate their family-centered care practices. The inventory is organized into 7 sections: (1) vision, mission, and philosophy of care; (2) family support; (3) information and decision making; (4) services coordination and continuity; (5) personnel practices; (6) quality improvement; and (7) community partnerships. Individual and group interviews were completed with a variety of staff in each emergency department. RESULTS: All 9 emergency departments demonstrated some integration of family-centered care principles. However, staff knowledge about family-centered care varied. Support of family-centered care was most consistent in the departments with specific competencies, educational programs, and practices that were inclusive of the family. DISCUSSION: The Family-centered Care Self-assessment Inventory tool was effective in evaluating family-centered care for pediatric patients in emergency departments. The assessment tool helped the departments to identify current family-centered care practices. Based on those assessments, the departments were able to identify areas of strength and opportunities for improvement in the care of children and their families. PMID- 11387561 TI - The case for a universal, valid, reliable 5-tier triage acuity scale for US emergency departments. PMID- 11387562 TI - A case management, education,and prevention program at a small emergency shelter for homeless men: one nurse's experience. PMID- 11387563 TI - HCFA's 250-yard rule for patients requiring assistance outside: EMS' or the hospital's responsibility? PMID- 11387564 TI - Firearm safety and children: access and attitudes. PMID- 11387565 TI - Health care workers enjoy some success in suing manufacturers of latex gloves. PMID- 11387566 TI - Violence in the emergency department: a firsthand account. PMID- 11387567 TI - EMSC and ENA introduce free periodically updated on-line pediatric emergency resource kit. PMID- 11387568 TI - Triaging pediatric orthopedic injuries. PMID- 11387569 TI - The clinical care and documentation for victims of drug-facilitated sexual assault. PMID- 11387570 TI - Antimicrobial central venous catheters: do they make a difference? PMID- 11387571 TI - A 68-year-old man with an acute painful swollen knee. PMID- 11387572 TI - Changes. PMID- 11387573 TI - Electric stimulation as an adjunct to heal diabetic foot ulcers: a randomized clinical trial. AB - OBJECTIVE: To evaluate high-voltage, pulse-galvanic electric stimulation as an adjunct to healing diabetic foot ulcers. DESIGN: Randomized, double-blind, placebo-controlled pilot trial. SETTING: University medical center. PATIENTS: Forty patients with diabetic foot ulcers, consecutively sampled. Twenty patients each assigned to treatment and placebo groups. Five patients (2 treated, 3 placebo) withdrew because of severe infection. INTERVENTIONS: Electric stimulation through a microcomputer every night for 8 hours. The placebo group used identical functioning units that delivered no current. Additional wound care consisted of weekly debridements, topical hydrogel, and off-loading with removable cast walkers. Patients were followed for 12 weeks or until healing, whichever occurred first. MAIN OUTCOME MEASURES: Proportion of wounds that healed during the study period. Compliance with use of device (in hr/wk), rate of wound healing, and time until healing. RESULTS: Sixty-five percent of the patients healed in the group treated with stimulation, whereas 35% healed with placebo (p = .058). After stratification by compliance, a significant difference was identified among compliant patients in the treatment group (71% healed), noncompliant patients in the treatment group (50% healed), compliant patients in the placebo group (39% healed), and noncompliant patients in the placebo group (29% healed, linear-by-linear association = 4.32, p = .038). There was no significant difference in compliance between the 2 groups. CONCLUSION: Electric simulation enhances wound healing when used in conjunction with appropriate off loading and local wound care. PMID- 11387574 TI - Physical activity in daily life in patients with chronic low back pain. AB - OBJECTIVES: To evaluate disuse (ie, a decreased daily physical activity level) in patients with chronic low back pain (LBP) and to evaluate the construct validity of accelerometry for measuring physical activity in daily life. DESIGN: Case control study in a cross-sectional design; comparison of accelerometry to the criterion standard (doubly labeled water technique). SETTING: Normal daily living (unrestricted by the measurement devices). PATIENTS: Thirteen patients with chronic nonspecific LBP and 13 age- and gender-matched healthy controls. MAIN OUTCOME MEASURES: Physical activity in daily life, expressed as whole-body acceleration measured with a triaxial accelerometer (Tracmor), and as the ratio between average daily metabolic rate (ADMR), measured by the doubly labeled water technique, and resting metabolic rate (RMR), measured by the ventilated hood. Both techniques were used simultaneously for 14 days. RESULTS: Mean physical activity level in patients and controls did not differ significantly. The correlation between the Tracmor and ADMR and RMR was .72 (p < .01). CONCLUSIONS: Decreased physical activity levels in this sample of chronic LBP patients was not confirmed. The Tracmor is a valid instrument for measuring daily activity in LBP patients. PMID- 11387575 TI - Back pain as a secondary disability in persons with lower limb amputations. AB - OBJECTIVE: To evaluate the frequency, duration, intensity, and interference of back pain in a sample of persons with lower limb amputations. DESIGN: Retrospective, cross-sectional survey. SETTING: Community-based survey from clinical databases. PARTICIPANTS: Participants who were 6 or more months post lower limb amputation (n = 255). INTERVENTION: An amputation pain survey that included several standardized pain measures. MAIN OUTCOME MEASURES: Frequency, duration, intensity, and interference of back pain. RESULTS: Of the participants who completed the survey (return rate, 56%), 52% reported experiencing persistent, bothersome back pain. Of these, 43% reported average back pain intensity in the mild range (1-4 on 0-10 rating scale) and 25% reported pain of moderate intensity (5-6 on 0-10 scale). Most respondents with back pain rated the interference of their pain on function as none to minimal. However, nearly 25% of those with back pain described it as frequent, of severe intensity (>or=7 on 0-10 scale), and as severely interfering with daily activities including social, recreational, family, and work activities. CONCLUSIONS: Back pain may be surprisingly common in persons with lower limb amputations, and, for some who experience it, may greatly interfere with function. PMID- 11387576 TI - Low back pain: reliability of a set of pain measurement tools. AB - OBJECTIVES: To determine the reliability and internal consistency of a set of low back pain (LBP)-related measurement tools and to determine whether they are appropriate for use in a large-scale, community-based sample in Israel. DESIGN: Test-retest reliability study, with an interval of 2 to 14 days between test and retest. SETTING: Physiotherapy clinics. PARTICIPANTS: One hundred fifty-one patients with LBP. MAIN OUTCOME MEASURES: The Modified Roland-Morris Disability Questionnaire (MRMQ); a simple verbal pain severity scale; and modified pain symptoms frequency and bothersomeness indices. Three measures of variables with potential association with LBP were also used: a Fear-Avoidance Beliefs Questionnaire (FABQ), work satisfaction scale, and the Baecke Physical Activity Questionnaire (BPAQ). RESULTS: Test-retest reliability was high for the MRMQ, pain symptom indices, work index of the FABQ, and occupational activity index of the BPAQ; the internal consistency of the MRMQ and FABQ work index were also high (intraclass correlation coefficient >or= .89; alpha = .89). CONCLUSIONS: Most measurement tools are reliable and suitable for community LBP studies in Israel. PMID- 11387577 TI - The role of unilateral spatial neglect in rehabilitation of right brain-damaged ischemic stroke patients: a matched comparison. AB - OBJECTIVE: To assess the specific influence of unilateral spatial neglect (USN) on rehabilitation outcome. DESIGN: A case-control study in consecutive stroke inpatients. SETTING: Rehabilitation hospital. PATIENTS: One hundred seventy-eight patients with sequelae of first stroke, enrolled in homogeneous subgroups, matched for age (within 1yr) and onset admission interval (within 3d), and difference because of the presence of USN, evaluated by a specific neuropsychologic battery. INTERVENTIONS: All patients received physical rehabilitation: physiotherapy for 60 minutes twice a day (once on Saturday), 6 days a week, within 24 hours of admission. USN-positive (USN(+)) patients received specific treatment of 5, 1-hour sessions per week, for 8 consecutive weeks: (1) visual scanning, (2) reading and copying, (3) copying of line drawings on a dot matrix, and (4) describing a scene. Patients were assessed with neurologic (Canadian Neurological Scale), neuropsychiatric (Hamilton Depression Rating Scale), neuroradiologic, and functional (Barthel Index, Rivermead Mobility Index [RMI]) examinations. MAIN OUTCOME MEASURES: Length of stay, efficiency (average daily increase in Barthel Index), effectiveness (amount of potential improvement achieved during rehabilitation) of treatment and percentage of low- and high-response patients calculated on the Barthel Index and the RMI, and percentage of urinary incontinence and return home were evaluated. Odds ratios (ORs) of dropouts and of low and high therapeutic response were also quantified. RESULTS: Compared with USN-negative patients, USN(+) patients had significantly more severe baseline neurologic and functional status at admission, less effectiveness and efficiency on activities of daily living (ADLs) and mobility, a higher percentage of low responders, longer hospitalization, a higher percentage of persistent incontinence at discharge (20.5% vs 4.9%), and a lower percentage of high responders and patients returning home. The presence of USN was incompatible with a high therapeutic response, for both ADLs (OR= 2.94, 95% confidence interval [CI]= 1.05-8.20; b +/- standard error = 1.08 +/- .52, p< .05), and mobility (OR = 7.16, 95% CI = 2.78-18.44; b = 1.97 +/- .52, p < .001) and was a relevant prognostic factor for institutional discharge (OR = 5.62, 95% CI = 1.63-19.38; b = 1.73 +/-.63, p < .01, accuracy 88.41%). CONCLUSIONS: The results of this study provide further strong evidence of the relationship between USN and disability in right brain-damaged patients and of its unfavorable impact on rehabilitation, despite the cognitive training performed by all USN(+) patients. PMID- 11387578 TI - The reliability of the wolf motor function test for assessing upper extremity function after stroke. AB - OBJECTIVE: To examine the reliability of the Wolf Motor Function Test (WMFT) for assessing upper extremity motor function in adults with hemiplegia. DESIGN: Interrater and test-retest reliability. SETTING: A clinical research laboratory at a university medical center. PATIENTS: A sample of convenience of 24 subjects with chronic hemiplegia (onset >1yr), showing moderate motor impairment. INTERVENTION: The WMFT includes 15 functional tasks. Performances were timed and rated by using a 6-point functional ability scale. The WMFT was administered to subjects twice with a 2-week interval between administrations. All test sessions were videotaped for scoring at a later time by blinded and trained experienced therapists. MAIN OUTCOME MEASURE: Interrater reliability was examined by using intraclass correlation coefficients and internal consistency by using Cronbach's alpha. RESULTS: Interrater reliability was.97 or greater for performance time and.88 or greater for functional ability. Internal consistency for test 1 was.92 for performance time and.92 for functional ability; for test 2, it was.86 for performance time and.92 for functional ability. Test-retest reliability was.90 for performance time and.95 for functional ability. Absolute scores for subjects were stable over the 2 test administrations. CONCLUSION: The WMFT is an instrument with high interrater reliability, internal consistency, test-retest reliability, and adequate stability. PMID- 11387579 TI - Comparing stimulation-induced pain during percutaneous (intramuscular) and transcutaneous neuromuscular electric stimulation for treating shoulder subluxation in hemiplegia. AB - OBJECTIVE: To determine whether percutaneous (intramuscular) neuromuscular electric stimulation (perc-NMES) is less painful than transcutaneous neuromuscular electric stimulation (trans-NMES) for treating shoulder subluxation in hemiplegia. DESIGN: Double-blind, crossover trial. SETTING: University affiliated tertiary care hospital. PARTICIPANTS: A convenience sample of 10 hemiplegic subjects with at least 1 fingerbreadth of glenohumeral subluxation. INTERVENTIONS: All subjects received 3 randomly ordered pairs of perc-NMES and trans-NMES to the supraspinatus and posterior deltoid muscles of the subluxated shoulder. Both types of stimulation were optimized to provide full joint reduction with minimal discomfort. MAIN OUTCOME MEASURES: Pain was assessed after each stimulation with a 10-cm visual analog scale (VAS) and the McGill Pain Questionnaire, using the Pain Rating Index (PRI) scoring method. Subjects were asked which type of stimulation they would prefer for 6 weeks of treatment. Wilcoxon's signed-rank test was used to compare median differences in VAS and PRI between perc-NMES and trans-NMES. RESULTS: Median VAS scores for perc-NMES and trans-NMES were 1 and 5.7, respectively (p = .007). Median PRI scores for perc NMES and trans-NMES were 7 and 19.5, respectively (p = .018). Nine of the 10 subjects preferred perc-NMES to trans-NMES for treatment. CONCLUSION: Data suggest that perc-NMES is less painful than trans-NMES in the treatment of shoulder subluxation in hemiplegia. PMID- 11387580 TI - Using early neuropsychologic testing to predict long-term productivity outcome from traumatic brain injury. AB - OBJECTIVE: To evaluate whether early neuropsychologic testing is useful in predicting long-term productivity outcome after traumatic brain injury (TBI). DESIGN: Validation cohort prediction study. SETTING: Four inpatient brain injury rehabilitation programs participating in the Traumatic Brain Injury Model Systems project. PARTICIPANTS: A total of 293 adults with nonpenetrating TBI. MAIN OUTCOME MEASURES: Fifteen neuropsychologic tests were administered to patients who emerged from posttraumatic amnesia before rehabilitation discharge. Test scores were classified in the normal range or impaired range, using objective criteria. Outcome was defined as productive if the patient was competitively employed or enrolled full time in regular education. RESULTS: Productivity at follow-up was predicted by completion of at least 1 neuropsychologic test before discharge, by an injury-test interval of less than 2 months, and by normal range scores on 10 of the 15 neuropsychologic tests. Normal range scores on these tests increased the probability of a productive outcome by 40% to 130%. CONCLUSIONS: Neuropsychologic testing can help predict long-term productivity even when performed before discharge from inpatient rehabilitation and at variable injury test intervals. Early testing should be interpreted in relation to injury-test interval. Because tests of multiple neuropsychologic domains predicted outcome, comprehensive evaluations might be more useful in predicting outcome. PMID- 11387581 TI - Computer-assisted training for improving wheelchair mobility in unilateral neglect patients. AB - OBJECTIVE: To investigate whether a computer-assisted training (CAT) program for patients with left unilateral neglect would decrease symptoms of this disorder. DESIGN: Case-control study. SETTING: Inpatient rehabilitation unit of a government medical center. PATIENTS: Twenty right-handed patients who showed left unilateral neglect on screening measures (Rey-Osterrieth Complex Figure, Random Letter Cancellation Test) were assigned to a CAT treatment group; and 20 patients who showed similar levels of unilateral neglect on the screening measures were assigned to a control group. INTERVENTIONS: All subjects were inpatients in an acute rehabilitation unit and received rehabilitation therapy, including physical and occupational therapy. The treatment group received the experimental, CAT program, 12 to 20 sessions of about 45 minutes each. Treatment consisted of 5 modules, each of increasing complexity, to improve attention to stimuli in the left hemisphere, and 2 simulated wheelchair obstacle courses to propel a wheelchair while avoiding obstacles. MAIN OUTCOME MEASURES: Computer tasks designed for this study (Video Tracking Test, Video Obstacle Course Test), a real life wheelchair obstacle course (WCOC), and incident reports indicating falls and accidents. RESULTS: Trained subjects performed significantly better on the WCOC than control subjects (F(1,36) = 23.41, p = .00003). Also, trained subjects had fewer incident reports than control subjects during their hospitalization (chi(2)(1,)(n)(=38) = 5.15, p = .023). CONCLUSIONS: CAT can reduce unilateral neglect symptoms on experimental tasks and some measures of accident risk. PMID- 11387582 TI - Rehabilitation of the geriatric vascular amputee patient: a population-based study. AB - OBJECTIVES: To determine the rate of successful prosthetic fitting in geriatric vascular amputees in the community and to determine predictors of successful fit. DESIGN: Epidemiologic survey. SETTING: General community, Olmsted County, Minnesota. PATIENTS: All Olmsted County residents more than 65 years old who had a major lower extremity amputation (below knee amputation [BKA] or higher) for peripheral vascular disease between 1974-1995, of whom 199 were identified. Median age at amputation was 79.7 years with a median survival of 1.5 years. INTERVENTION: A retrospective chart review. MAIN OUTCOME MEASURE: Successful prosthetic fit. RESULTS: Amputation levels were: 64% BKA, 4.5% knee disarticulation, 31% above knee amputation (AKA), and 0.5% hip disarticulation. Only 36% of the population was successfully fitted, compared with 74% of patients referred to the Amputee Clinic. Major reasons for not being fitted included death, reamputation, cerebrovascular disease, and cognitive deficits. Increased age (p < .001), cerebrovascular disease (p < .001), dementia (p = .002), and AKA (p < .001) were associated with failure to fit. CONCLUSION: The high probability of successful prosthetic fitting reported among referral practices cannot be generalized to unselected elderly individuals. However, selected individuals can successfully be fitted with a prosthesis; knowledge of predictors of prosthetic fitting may facilitate setting of realistic goals during presurgical counseling in this age group. PMID- 11387583 TI - Utility of a composite measure to detect problematic alcohol use in persons with traumatic brain injury. AB - OBJECTIVES: To examine factors complicating the study of alcohol-related effects in traumatic brain injury (TBI) patients and to evaluate a composite measure to categorize such patients according to degree of alcohol-related problems. DESIGN: Inception cohort. SETTING: Level I trauma center. PATIENTS: Consecutively hospitalized adult TBI patients (n = 156; 73% men; 87% Caucasian; mean age, 30yr; mean education, 12yr). Selection criteria required objective evidence of brain trauma; minimum survival of 1 month postinjury; age 15 years or older; and English speaking. MAIN OUTCOME MEASURES: An index of problematic drinking based on a measure created by combining blood-alcohol level, quantity-frequency of consumption, and the Short Michigan Alcoholism Screening Test. Preinjury characteristics were obtained through structured interview. RESULTS: Participants with highly problematic drinking showed poorer premorbid psychosocial functioning, including lower educational attainment, greater likelihood of problems with the law, lower perceived social support, and greater prevalence of other substance abuse. CONCLUSION: The composite index is useful in identifying problematic drinkers among TBI patients. Results have implications for interpreting and planning research on the role of alcohol in TBI outcomes. PMID- 11387584 TI - Wellness perception in persons with traumatic brain injury and its relation to functional independence. AB - OBJECTIVES: To test the reliability and validity of a perceptual wellness measure in persons after traumatic brain injury (TBI) and to determine whether a relation exists between functional independence and wellness perceptions in the same population. DESIGN: Survey research. SETTING: A private, residential brain injury program. PARTICIPANTS: A convenience sample of 49 patients (43 men, 6 women) with TBI whose mean age was 32.1 years (range, 18-61yr) and mean time since injury was 10.47 years (range, 1-21yr). MAIN OUTCOME MEASURES: The Perceived Wellness Survey (PWS) assessed wellness. The PWS has 6 subscales measuring physical, psychologic, emotional, intellectual, spiritual, and social wellness. The FIM instrument was used to measure functional status. RESULTS: The mean PWS score (15.99) for the sample was comparable to published samples of adults (mean, 15.31-16.51); however, the reliability of the composite score (alpha = .58) and the subscales (alpha = .32-.64) was less than that obtained in previous samples (composite alpha = 0.91; subscale alpha = .64-.81). The correlations between the PWS and the FIM scores were not significant. CONCLUSIONS: The PWS in its composite form is a reliable measure for use with persons with TBI. The finding that perceived wellness and functional independence were not related suggests that these constructs are unique and thus should both be measured. The measurement of perceptions will enable the provider to consider a client holistically and to develop programs that address quality of life issues. Further, because perceptions influence behaviors, understanding a person's perceptions in multiple dimensions may provide a useful and necessary framework for developing intervention programs that address behavioral and cognitive issues that are important to that person. PMID- 11387585 TI - Rehabilitation of postural disturbances of hemiplegic patients by using trunk control retraining during exploratory exercises. AB - OBJECTIVE: To assess use of the Bon Saint Come device for axial postural rehabilitation in hemiplegic patients, a technique based on voluntary trunk control during exploratory retraining. DESIGN: A 3-month randomized controlled trial. SETTING: A public neurorehabilitation center. PATIENTS: Twenty consecutive hemiplegic patients with axial postural disturbance resulting from recent stroke were randomly assigned to a device group (DG) or control group (CG). The 2 groups of 10 patients were similar. INTERVENTION: For 1 month, the DG patients followed an experimental program for 1 hour daily and conventional neurorehabilitation for 1 hour daily, whereas CG patients had conventional neurorehabilitation for 2 hours daily. For the next 2 months, all 20 patients had conventional neurorehabilitation for 2 hours daily. MAIN OUTCOME MEASURES: Patients were assessed on days 0, 30, and 90 by using a battery of postural tests, gait evaluation, the Bells neglect test, and the FIM instrument. RESULTS: On day 30, postural and neglect tests improved significantly more in DG than in CG. The benefit remained at day 90. Gait improved earlier in DG than in CG. FIM scores improved equally. CONCLUSIONS: Voluntary trunk control retraining during spatial exploration with the Bon Saint Come device appears to be a useful approach for rehabilitation of postural disorders in hemiplegic patients. Treatments designed to improve spatial cognition deficits probably enhance postural disorder recovery in hemiplegia. PMID- 11387586 TI - Self-reported exercise before age 40: influence on quantitative skeletal ultrasound and fall risk in the elderly. AB - OBJECTIVES: To compare musculoskeletal factors with bone structure, as measured by quantitative ultrasound (QUS) at the calcaneus, and their potential to predict fall risk in geriatric inpatients. DESIGN: Longitudinal. SETTING: Two geriatric hospitals in Switzerland. PARTICIPANTS: A total of 134 of 207 long-stay geriatric patients (96 women, 38 men) who were able to perform the timed up and go (TUG) test. INTERVENTIONS: Five musculoskeletal tests: 2 functional tests (TUG, for mobility; functional reach test, for balance), and 3 muscle strength tests (knee flexor, knee extensor, grip). Falls were monitored prospectively in a subgroup of 94 mobile subjects of 1 geriatric hospital throughout each individual length of stay (median, 31.4wk: interquartile range, 16-56.4wk). MAIN OUTCOME MEASUREMENTS: Functional and strength tests, mobility status, and self-reported exercise before age 40 were musculoskeletal factors to be compared with QUS. RESULTS: QUS was higher in mobile subjects without walking aid (p < .0001) and correlated significantly with muscle strength (knee flexor: r = .36; knee extensor: r = .30) and functional tests (TUG: r = -.25; functional reach: r = .16). Women who reported regular exercise before age 40 had higher QUS (p = .01) and fewer falls (p = .01). Falls were less frequent in subjects with walking aid (p = .03). No single musculoskeletal test, but rather a combination of demographic variables, musculoskeletal factors, and QUS could predict 76% of total variation of fall risk. CONCLUSION: This study showed the important impact of current mobility and muscle strength status on bone structure, as measured by QUS at the calcaneus. In addition, a beneficial effect of former exercise on QUS and fall risk at advanced age could be documented in women. Both findings support life-long engagement in exercise, which might be particularly meaningful for women. PMID- 11387587 TI - A validation of the 10-meter incremental shuttle walk test as a measure of aerobic power in cardiac and rheumatoid arthritis patients. AB - OBJECTIVE: To validate a simple, clinically relevant, and inexpensive test of aerobic power-the 10-meter incremental shuttle walk test (SWT)--in 2 separate patient populations. DESIGN: Two-sample validity study. SETTING: Physiotherapy department of major hospital in the United Kingdom. PATIENTS: Convenience samples of rheumatoid arthritis (RA) patients (n = 10) and cardiac patients (n = 10). INTERVENTION: Subjects were attached to a portable respiratory gas analyzer to measure oxygen uptake. They walked around an oval 10-meter course, starting at 0.5m/s, with velocity gradually increased by .17m/s increments for as long as they could, for up to 12 minutes. MAIN OUTCOME MEASURES: A subject's maximal rate of oxygen uptake during exercise (V(O)(2)max) established with linear extrapolation was regressed against the number of shuttles completed (distance walked). An earlier study (n = 28) showed high levels of reliability and validity with linear extrapolation. RESULTS: No significant linear relationship was found between V(O)(2)max and the number of shuttles completed (R(2); RA subjects = 9.7%, cardiac subjects = .03%, p > .05). CONCLUSION: These results do not support use of the SWT as a representative measure of aerobic power. Despite this finding, the advantages of developing a clinically viable alternative to costly laboratory testing warrants further study of the SWT in patient groups. PMID- 11387588 TI - Physical exercise and/or enriched foods for functional improvement in frail, independently living elderly: a randomized controlled trial. AB - OBJECTIVE: To examine the effects of an exercise program and an enriched food regimen on physical functioning of frail elderly persons. DESIGN: A 17-week randomized, placebo-controlled trial. SETTING: Community. PARTICIPANTS: One hundred fifty-seven independently living frail elderly (mean age, 78.7 +/- 5.6yr). INTERVENTION: Thirty-nine subjects participated in a twice weekly group exercise designed to improve daily functioning; 39 subjects daily ate foods enriched with vitamins and minerals (at 25%-100% of the recommended daily allowances); 42 subjects exercised and ate enriched foods; and 37 subjects served as controls. Nonexercising groups followed a social program; nonsupplement groups received the same food products without the micronutrients. MAIN OUTCOME MEASURES: Functional performance based on 6 performance tests, physical fitness based on 7 fitness tests, and disabilities based on the self-reported ability to perform 16 daily activities. RESULTS: Performance sum scores were significantly enhanced in trained (+8%) compared with nontrained subjects (-8%) (difference in change: 1.9 points, p < .001, adjusted for baseline scores). Fitness sum scores were significantly enhanced as well (+3% in trained vs -2% in nontrained) (difference in change: 0.9 points, p = .05, adjusted for baseline scores). No exercise effects on the disability score were observed. Consumption of enriched products did not affect performance, fitness, or disability scores. CONCLUSION: Our comprehensive exercise program, designed for widespread applicability, enhanced physical performance and fitness in a population of frail elderly. Daily consumption of micronutrient enriched foods showed no functional benefits within 17 weeks. PMID- 11387589 TI - Combined use of body weight support, functional electric stimulation, and treadmill training to improve walking ability in individuals with chronic incomplete spinal cord injury. AB - OBJECTIVE: To assess the effect of an intervention combining body weight support (BWS), functional electric stimulation (FES), and treadmill training on overground walking speed (OGWS), treadmill walking speed, speed and distance, and lower extremity motor scores (LEMS). DESIGN: Before and after comparison. SETTING: Miami Project to Cure Paralysis. PARTICIPANTS: Nineteen subjects with American Spinal Injury Association class C injury who were at least 1 year postinjury and had asymmetrical lower extremity function. INTERVENTION: Subjects trained 1.5 hours per day, 3 days per week, for 3 months. The training consisted of body weight-supported treadmill walking assisted by electric stimulation. Stimulation was applied to common peroneal nerve of the weaker lower extremity (LE) and timed to assist with the swing phase of the step cycle. MAIN OUTCOME MEASURES: OGWS in the absence of both BWS and FES; LEMS, and treadmill training parameters of speed and distance. RESULTS: Over the course of training, there was a significant increase in OGWS (from.12 +/- 0.8m/s to .21 +/- .15m/s, p = .0008), treadmill walking speed (from .23 +/- .12m/s to.49 +/- .20m/s, p = .00003), and treadmill walking distance (from 93 +/- 84m to 243 +/- 139m, p = .000001). The median LEMS increased significantly for both the stimulated and nonstimulated leg (from 8 to 11 in the FES-assisted leg, from 15 to 18 in the nonassisted leg, p < .005 for each). CONCLUSIONS: All subjects showed improvement in OGWS and overall LE strength. Further research is required to delineate the essential elements of these particular training strategies. PMID- 11387590 TI - Supported treadmill ambulation training after spinal cord injury: a pilot study. AB - OBJECTIVES: To conduct a pilot study of weight-supported ambulation training after incomplete spinal cord injury (SCI), and to assess its safety. DESIGN: Quasiexperimental, repeated measures, single group. SETTING: Veterans Affairs medical center. PATIENTS: Three subjects with incomplete, chronic, thoracic SCIs; 2 classified as D on the American Spinal Injury Association (ASIA) impairment scale and 1 as ASIA impairment scale C. INTERVENTION: Subjects participated in 12 weeks of training assisted by 2 physical therapists. The training consisted of walking on a treadmill while supported by a harness and a pneumatic suspension device. Support started at 40% of body weight and a treadmill speed of.16kmph, and progressed by reducing support and increasing treadmill speed and continuous treadmill walking time up to 20 minutes. Training was conducted for 1 hour per day, 5 days per week for 3 months. Treadmill walking occurred for 20 minutes during the sessions. MAIN OUTCOME MEASURES: Gait function (speed, endurance, walking status, use of assistive device and orthotics); oxygen costs of walking; brain motor control assessment; self-report indices; ASIA classification; muscle function test; and safety. RESULTS: All 3 subjects increased gait speed (.118m/s initially to.318m/s after training 12wk), and gait endurance (20.3m/5min initially to 63.5m/5min). The oxygen costs decreased from 1.96 to 1.33mL x kg(-1) x m(-1) after 12 weeks of training. CONCLUSIONS: This pilot study suggests that supported treadmill ambulation training can improve gait for individuals with incomplete SCIs by using objective gait measures. The self-report indices used have promise as patient-centered outcome measures of this new form of gait training. A larger, controlled study of this technique is warranted. PMID- 11387591 TI - Peripheral vascular changes after electrically stimulated cycle training in people with spinal cord injury. AB - OBJECTIVE: To test whether a short period of training leads to adaptations in the cross-sectional area of large conduit arteries and improved blood flow to the paralyzed legs of individuals with spinal cord injury (SCI). DESIGN: Before-after trial. SETTING: Rehabilitation center, academic medical center. PARTICIPANTS: Nine men with spinal cord lesions. INTERVENTION: Six weeks of cycling using a functional electrically stimulated leg cycle ergometer (FES-LCE). MAIN OUTCOME MEASURES: Longitudinal images and simultaneous velocity spectra were measured in the common carotid (CA) and femoral (FA) arteries using quantitative duplex Doppler ultrasound examination. Arterial diameters, peak systolic inflow volumes (PSIVs), mean inflow volumes (MIVs), and a velocity index (VI), representing the peripheral resistance, were obtained at rest. PSIVs and VI were obtained during 3 minutes of hyperemia following 20 minutes of FA occlusion. RESULTS: Training resulted in significant increases in diameter (p < .01), PSIVs (p < .01), and MIVs (p < .05), and reduced VI (p < .01) of the FA, whereas values in the CA remained unchanged. Postocclusive hyperemic responses were augmented, indicated by significantly higher PSIVs (p <.01) and a trend toward lower VI. CONCLUSION: Six weeks of FES-LCE training increased the cross-sectional area of large conduit arteries and improved blood flow to the paralyzed legs of individuals with SCI. PMID- 11387592 TI - Variation in shoulder position sense at mid and extreme range of motion. AB - OBJECTIVE: To determine the effect of different joint positions on position sense of asymptomatic shoulders. DESIGN: Repeated-measures design. SETTING: Laboratory in an educational institution. PARTICIPANTS: Thirty-four asymptomatic, right handed men. INTERVENTIONS: The ability of subjects to replicate 3 criterion positions was examined on subjects' right shoulders by using an isokinetic dynamometer. Three criterion positions were the 50th, 75th, and 90th percentiles of each individual's total passive shoulder rotation range measured from the full internal rotation position. MAIN OUTCOME MEASURE: Repositioning accuracy, indicating the difference in degrees between the criterion and matching positions. RESULTS: All subjects were able to reproduce the criterion position near the end of external rotation range more accurately and consistently than in the middle range of motion (ROM). CONCLUSIONS: Position sense acuity at the shoulder complex varied across the ROM and may be enhanced near the end of rotation range where there is more tension on the restraints to movement. Therefore, an individual's ROM should be factored into any attempt to assess or rehabilitate shoulder proprioception. PMID- 11387593 TI - The relationship between intramuscular temperature, skin temperature, and adipose thickness during cryotherapy and rewarming. AB - OBJECTIVE: To describe the relationships among muscle temperature, skin temperature, room temperature, body core temperature, time, and subcutaneous adipose thickness during cryotherapy and rewarming. DESIGN: A multiple linear regression with 5 independent variables (skin temperature, body core temperature, subcutaneous adipose thickness, room temperature, time) predicting intramuscular (IM) temperature. SETTING: A sports injury research laboratory. PARTICIPANTS: Fifteen volunteers with thigh skinfold measurements smaller than 40 mm. INTERVENTIONS: Thirty-minute cryotherapy treatment (ice bag) followed by a 120 minute rewarming period. MAIN OUTCOME MEASURES: The relationship between skin and IM temperature was described, and an equation predicting IM temperature by using room temperature, skin temperature, body core temperature, time, and adipose thickness was developed. RESULTS: Pearson's correlations between each predictor variable of IM temperature during cryotherapy were skin temperature, r = .46; skinfold, r = .37; time, r = -.59; core temperature, r =.21; and room temperature, r = -.47. During rewarming, the correlations were skin temperature, r = .71; skinfold, r = .27; time, r = .76; core temperature, r = - .05; and room temperature, r = - .21. A multiple regression equation (R(2) = .76) was developed to predict IM temperature during cryotherapy. A separate equation (R(2) =.81) was developed to predict muscle temperatures during rewarming. CONCLUSIONS: During and after ice application, no single predictor adequately explained the change in IM temperature. Skin surface temperature was a weak predictor of IM temperature during cryotherapy and should not be used as the sole dependent measure in cryotherapy efficacy studies. PMID- 11387594 TI - Complex regional pain syndrome type I treated with topical capsaicin: a case report. AB - This report describes the case of a multitrauma patient who underwent an amputation of the left arm and had a complicated left crural fracture with a delayed union. He was treated in an inpatient setting for preprosthetic training for a myoelectric prosthesis and to regain walking abilities. After consolidation of the crural fracture, complex regional pain syndrome type I (CRPS I) developed in the left foreleg, which hindered mobilization. Topical capsaicin .075% was prescribed and a stress-loading mobilization schema was instituted. No other treatment modalities directed at CRPS I were added. After 6 weeks, no signs or symptoms of CRPS I were present and capsaicin was discontinued. Capsaicin is a well-accepted and documented treatment modality in neuropathic pain states such as postherpetic neuralgia. However, it has rarely been described in CRPS I. Capsaicin is discussed within the framework of recent insights in the neurobiology of nociception, and it is concluded that it may provide a theory driven treatment for CRPS I, especially in the acute stage, that facilitates physical therapy and prevents peripheral and spinal sensitization. PMID- 11387595 TI - Intrathecal baclofen for generalized dystonia: a case report. AB - Dystonia is an abnormal movement characterized by sustained muscle contractions, frequently causing twisting and repetitive movements or abnormal postures. Primary generalized dystonia usually evolves to a debilitating condition, often unresponsive to oral pharmacologic regimens or physical rehabilitation. We present the case of a patient with idiopathic adult-onset generalized dystonia who was recalcitrant to oral therapies. This patient was treated with intrathecal baclofen therapy and responded dramatically with improved functional abilities, resolution of chronic pain, and an improved rating on the dystonia scale. PMID- 11387596 TI - Evaluation of effect of ankle-foot orthosis use on strength restoration of paretic dorsiflexors. AB - OBJECTIVE: To assess whether a difference exists in restoration of strength between patients with a recent paresis of the dorsiflexors of the ankle using an ankle-foot orthosis (AFO) and patients without an AFO. DESIGN: Prospective case control study. SETTING: Patients from regional hospitals, tested in a rehabilitation research center. PARTICIPANTS: Twenty-nine patients with a recent (6wk-1yr) peripheral paresis, alternately assigned to a group using an AFO or a control group. There was no significant difference in duration of the paresis and in torque at entering the study (T0) between the 2 groups. INTERVENTIONS: The use of an ankle-foot orthosis. MAIN OUTCOME MEASURES: Isometric torque production of ankle dorsiflexors, expressed as ratio of paretic and healthy side, in 2 measurement sessions, over a period of 6 weeks (T0-T6) with the ankle in 0 degrees and 30 degrees plantarflexion. RESULTS: Both groups had significant restoration of strength +/- standard deviation between T0 and T6 in 30 degrees flexion: non-AFO group 17% +/- 15%, AFO group 9% +/- 12%. No significant difference existed between the 2 groups (30 degrees p = .56). No significant shift in strength ratio 0 degrees :30 degrees occurred (AFO group p = .82). CONCLUSION: The use of an orthosis does not influence restoration of strength in patients with a recent peripheral paresis of the ankle dorsiflexors. PMID- 11387597 TI - Comparative trial of stent-like balloon angioplasty versus coronary stenting for acute myocardial infarction. AB - Primary PTCA has been shown to be superior to any thrombolytic regimen and offers higher reperfusion rates and better coronary flow grades. Its limitations include recurrent ischemia (10%-15%), infarct-related artery reocclusion (5%-10%), angiographic restenosis (35%-50%), and need to perform repeat PTCA or CABG at 6 month follow-up (20%). Thus, the current role of coronary stenting for acute myocardial infarction (AMI) is very promising. From December 1995 through January 1997, 335 patients underwent primary angioplasty during the first 12 hr from symptom onset at our institution. We performed a retrospective study comparing the in-hospital and 6-month follow-up outcome of 61 patients who underwent coronary stenting (stent group) against 61 patients with optimal (residual lesion stenosis < 30%) balloon-only primary angioplasty (stent-like group). Patients were routinely treated with aspirin, and ticlopidine was given only to the stent group. In-hospital major adverse cardiac events (MACE) rate was 11.5% without statistical differences between the groups. Cardiac death rate was similar in both groups (4.9 vs. 6.6%; P = 1.0) and only two (3.3%) patients from the stent group and none from the PTCA group had nonfatal myocardial reinfarction. At 6 month follow-up, the rate of recurrent angina was higher in the stent-like group (30.9 vs. 7.1%; P < 0.001). Multivariate analysis showed that only stenting of the infarct-related artery was a borderline independent predictor for MACE (OR = 0; 95% CI = 0-1; P = 0.057). Primary stenting for AMI reduces the rate of recurrent angina or symptoms and MACE at 6-month follow-up. PMID- 11387598 TI - Late (> 48 hr) myocardial infarction after PTCA: clinical and angiographic characteristics of infarction related or not to the angioplasty site. AB - Since late myocardial infarctions after percutaneous coronary interventions have not been well characterized, we intended to evaluate the characteristics of myocardial infarctions occurring > 48 hr after balloon angioplasty of native coronary arteries or saphenous vein grafts. The Montreal Heart Institute database (1985-1996) was interrogated for all patients readmitted with a diagnosis of MI more than 48 hr after successful percutaneous transluminal coronary angioplasty (PTCA). We compared the clinical, procedural, and angiographic variables between MIs related or not to the index PTCA site. One hundred and ninety-three patients presented with late myocardial infarction (MI) following balloon angioplasty. The median time elapsed between PTCA and MI was 55 days compared to 968 days when MI was unrelated to the PTCA site. MIs related to the PTCA site were more likely non Q-wave (76% vs. 35%, P = 0.0001) with less marked CK-MB rise. Angiography showed less complex lesions (63% vs. 90%, P = 0.001) and better thrombolysis in myocardial infarction (TIMI) grade flow (TIMI II to III, 66% vs. 56%, P = 0.01) when the culprit lesion was at the PTCA site. Independent predictors of MI at the PTCA site were vein graft dilation, female sex, and residual stenosis post-PTCA. Myocardial infarctions occurring late after PTCA have a distinct time course and present specific characteristics according to their relationship or not to the previously dilated site. PMID- 11387599 TI - Suture closure of the femoral arteriotomy following invasive cardiac procedures: a detailed analysis of efficacy, complications, and the impact of early ambulation in 1,200 consecutive, unselected cases. AB - The purpose of this study was to assess the efficacy and safety of using a percutaneous suture device to close femoral arteriotomies following invasive cardiac procedures. All patients presenting for invasive cardiac procedures performed from the femoral artery were considered for suture closure. Patients were carefully assessed for access site complications, oozing, and the impact of suture closure on the safety of early ambulation. Clinical follow-up at 3-6 months was performed to assess for late complications. Femoral artery suture closures were performed in 1,200 consecutive cases in 1,097 patients. In 12.8% of cases, the patients ambulated within 1 hr. The success rate was 91.2% and the complication rate was 3.4%. Complications included the development of a hematoma (2.1%), the need for vascular surgery (0.6%), retroperitoneal hemorrhage (0.3%), blood transfusion (0.7%), local infection (0.5%), and pseudoaneurysm formation (0.1%). Factors found to be independently predictive of procedural failure were an age > 70 years, an ACT > 300 sec, left femoral artery access, and the performance of primary angioplasty. Follow-up at 3-6 months revealed no major hemorrhagic complications. We conclude that percutaneous suture closure effectively achieves femoral artery hemostasis in patients undergoing invasive cardiac procedures. The technique permits early ambulation and is associated with a relatively low incidence of complication. PMID- 11387600 TI - Postinterventional transcutaneous suture of femoral artery access sites in patients with peripheral arterial occlusive disease: a study of 930 patients. AB - The purpose of this study was to evaluate a percutaneous vascular suture (PVS) device in patients with peripheral arterial occlusive disease (PAOD) for achievement of immediate hemostasis at the vascular access site and early ambulation of fully anticoagulated patients after peripheral interventional procedures. From June 1995 to March 2000, a vascular suture using a PVS device (6 10 Fr) was applied in 930 patients with PAOD. All patients had received an endoluminal intervention in the pelvic and/or the contralateral femoropopliteal region via a retrograde access through the common femoral artery (CFA). The incidence of complications within 12 hr after intervention, prior to discharge, and at 30-day follow-up was assessed employing clinical examination, treadmill test, and color Doppler ultrasound and the safety of the PVS device was determined. The efficacy of the system was measured by the percentage of achieved immediate hemostasis and early ambulation. PVS was technically successful in 92.2% independently from the degree of calcification at the access site. In 7.8%, an appropriate suture delivery could not be performed and these patients were successfully treated by conventional compression technique. Device malfunction or insufficient suture closure occurred in 1.7% and 2.1%, respectively. In 7.0%, groin-related complications occurred. Ambulation within 2-4 hr after successful suture was possible in 96.1%. All patients were free of any local symptoms at 30 day follow-up. The PVS device provides a safe and effective solution to achieve immediate hemostasis, thus permitting early ambulation in fully anticoagulated patients with PAOD after peripheral interventional procedures. PMID- 11387601 TI - Acute and mid-term results of phosphorylcholine-coated stents in primary coronary stenting for acute myocardial infarction. AB - The aim of this pilot study was to evaluate the safety and efficacy of the BiodivYsio phosphorylcholine-coated stent in the primary treatment of acute myocardial infarction. The BiodivYsio stent (Biocompatible) is a balloon expandable stent, laser etched from a 316 L stainless steel tube. This device is coated with phosphorylcholine, a synthetic, hemocompatible phospholipid polymer that has been shown in experimental studies to reduce platelet and protein adhesion to the surface of the metal. One hundred consecutive patients within 24 hr of symptoms of onset of acute MI, treated with primary PTCA, were enrolled. After PTCA, stenting was attempted in all eligible lesions (reference diameter > or = 2.5 mm; no bend lesion > 45 degrees ). Poststenting regimens contained ticlopidine (500 mg/day) and aspirin (325 mg/day) and 6-12 hr of heparin infusion. Procedural success (TIMI > or = II and residual stenosis < 30%) was obtained in 70/74 cases (95%). TIMI grade III was restored in 90% of cases. In the patient group with procedural success (70 cases), 70 BiodivYsio stents were placed. After stenting, diameter stenosis decreased from 96% +/- 11% to 22% +/- 12% (P < 0.01) and minimal luminal diameter increased from 0.13 +/- 0.29 to 2.47 +/- 0.43 (P < 0.01). Nominal stent diameter was between 3.0 and 4.0 mm (mean, 3.5 +/- 0.4 mm). Stent length was between 11 and 28 mm (mean, 17 +/- 4.5 mm). Clinical follow-up was obtained in all patients; angiographic follow-up was performed in 65/70 (93%). No acute or subacute thrombosis was reported. Two in hospital major adverse cardiac events (MACE) were reported due to a nontreated left main disease that required coronary artery bypass graft (CABG) surgery. At follow-up, MACE were found in 9 of 68 patients (13%), target lesion revascularization (TLR) in 6%, and CABG in the remaining 6%. Primary stenting with phosphorylcholine-coated stent leads to excellent short- and mid-term clinical outcomes and is associated with a restenosis rate of 12%. PMID- 11387602 TI - Minor myocardial injury after elective uncomplicated successful PTCA with or without stenting: detection by cardiac troponins. AB - Cardiac troponins are sensitive and specific markers for the detection of minor myocardial injury. However, they have been rarely used to monitor myocardial injury after coronary stenting. The purpose of the study was to measure cardiac troponin I (cTnI) and cardiac troponin T (cTnT) levels after elective uncomplicated successful percutaneous transluminal coronary angioplasty (PTCA) with or without coronary stenting and to compare their results with serum creatinine kinase MB isoenzyme (CKMB). CTnI and cTnT levels were compared with those of CK or CKMB in 98 consecutive patients with stable angina undergoing elective uncomplicated successful PTCA with stenting (n = 71) or without stenting (n = 27). Markers were measured before and 6, 12, 24, and 48 hr after the procedure. Peak postprocedural levels for each marker were compared and related to angiographic and procedural characteristics as well as to the occurrence of side-branch occlusion. None of the patients had abnormal markers before the procedure. Abnormal postprocedural values of one or more markers were observed in 28 patients (29%), 23 after stenting and 5 after PTCA alone. The frequencies of abnormal cTnI and cTnT levels were significantly higher than that of CKMB after coronary intervention (26% and 18% vs. 7%; P = 0.00016 and 0.015, respectively), with cTnI being the most significant. When compared with troponin-negative patients, abnormal cardiac troponin values were significantly related to total time of inflation (223 +/- 128 vs. 170 +/- 105 sec; P = 0.008) and inflation maximal pressure (12.9 +/- 2.3 vs. 12.0 +/- 2.7 atm; P = 0.04). Small side-branch occlusion was noticed in 36% of the troponin-positive patients and in 6% of the troponin-negative group (P = 0.00047). In conclusion, minor myocardial injury is not uncommon after elective uncomplicated successful PTCA with or without stenting. Cardiac troponins, especially cTnI, are more sensitive than CKMB for the detection of this minor myocardial injury. Total time of inflation and inflation maximal pressure are predictors of postprocedural elevation of cardiac troponins. Side-branch occlusion may account for some, but not all, periprocedural minor myocardial injury. PMID- 11387603 TI - Comparison of relative coronary Doppler flow velocity reserve to stress myocardial perfusion imaging in patients with coronary artery disease. AB - To compare relative coronary artery vasodilator reserve (rCVR = CVRtarget/CVRreference) to myocardial perfusion stress imaging, 48 patients with coronary artery stenoses (61% +/- 16%; mean, +/- SD; range, 30%-91%) had measurements of target and reference vessel CVR (Doppler-tipped guidewire). rCVR was computed and compared to stress 201thallium or (99m)technetium-sestamibi myocardial tomography. Compared to 24 patients with negative stress imaging studies, 24 patients with positive stress studies had angiographically more severe stenoses (74% +/- 13% vs. 44% +/- 24%; P = 0.0005) with lower CVR(target) (1.68 +/- 0.55 vs. 2.46 +/- 0.74; P = 0.002) and lower rCVR (0.72 +/- 0.22 vs. 1.0 +/- 0.26; P < 0.003). Based on receiver-operator characteristic (ROC) cut points (CVR > 1.9; rCVR > 0.75), compared to CVR, rCVR had similar agreement (Kappa 0.54 vs. 0.50), sensitivity (63% vs. 71%), specificity (88% vs. 83%), and positive predictive value (83% vs. 81%) with myocardial perfusion tomography. A concordant CVRtarget/rCVR only slightly increased sensitivity, specificity, and positive predictive values (77%, 90%, and 87%, respectively). Although rCVR, like CVR, correlates with stress myocardial perfusion imaging results, rCVR did not have significant incremental prognostic value over CVR alone for myocardial perfusion imaging. However, rCVR does provide additional information regarding the status of the microcirculation in patients with coronary artery disease and complements the CVR for lesion assessment. PMID- 11387604 TI - How to define coronary stenosis severity? PMID- 11387605 TI - Actual outpatient PTCA: results of the OUTCLAS pilot study. AB - This study tested the safety and feasibility of coronary angioplasty on an outpatient basis. The purpose of this approach includes cost-effectiveness and patient comfort. Included were 159 patients treated with balloon angioplasty or intracoronary stent placement, all performed via the radial artery with 6 French guiding catheters. Patients were selected for same-day discharge based on the absence of any adverse predictor for subacute occlusion or unfavorable clinical outcome during the first 24 hr after successful PTCA. One hundred and six (66%) patients were discharged 4-6 hr after PTCA. Stents were used in 40% of patients. There were no cardiac or vascular complications. We conclude that outpatient PTCA, performed via the radial artery, is both safe and feasible in a large part of a routine PTCA population. PMID- 11387606 TI - Optimizing safe femoral access during cardiac catheterization. AB - This article describes the natural variability of femoral arterial anatomy as seen by angiography in a standard position. An analysis of 60 consecutive peripheral angiograms revealed data on average vessel diameter, variability in that diameter and the level of bifurcation of the common femoral artery into its two main branches. The statistical likelihood of optimal femoral arterial access through a standard dermotomy placed midway between, and 3 cm below, the imaginary line extending from the superior anterior iliac spine to the pubic symphysis was calculated. Our research disclosed average vessel diameters of 6.6 mm (3.9-8.9 mm), 5.2 mm (2.5-9.6 mm), and 4.9 mm (2.7-7.6 mm) for the common femoral artery, superficial femoral artery and deep femoral artery, respectively. The bifurcation of the common femoral artery into its branches averaged 5.5 mm below the inferior margin of the femoral head in most cases (76% of the time). In our laboratory, the average distance from the inferior border of the femoral head to our actual arteriotomy was 14.0 mm. Using our standard dermotomy, we achieved this favorable access position in 97% of cases. PMID- 11387608 TI - Randomized comparison of coronary angiography using 4F catheters: 4F manual versus "Acisted" power injection technique. AB - Compared with 6F catheters, diagnostic coronary angiographic and ventriculographic images with 4F catheters can be obtained with equivalent results using less radiographic contrast volume. Whether 4F coronary angiography would be superior using a power-assisted, operator-controlled technique compared with manual technique is unknown. To determine whether 4F coronary angiography using operator-controlled power injection (Acist, Minneapolis, MN) was equivalent or superior to the 4F manual technique, 96 unselected patients undergoing transfemoral coronary angiography were randomized to 4F catheter using a power injection or manual technique. Procedural characteristics and angiographic quality scores were analyzed. Comparing the 4F manual with the 4F power-injection technique, coronary angiographic quality scores were equivalent (left coronary artery 4.7 +/- 0.5 vs. 4.7 +/- 0.6, P = 0.99; right coronary artery 4.94 +/- 0.2 vs. 4.88 +/- 0.1, P = 0.21). Left ventriculography scores were lower in 4F Acist with similar contrast volumes. The total study contrast volume was significantly less in the 4F Acist group (119 +/- 35 vs. 149 +/- 49 ml, P = 0.001). Compared with the 4F manual contrast injection technique, diagnostic angiography through 4F catheters with power contrast injection resulted in equivalent coronary angiographic image quality with significantly less radiographic contrast volume. PMID- 11387607 TI - Coronary angioplasty and Rotablator atherectomy trial (CARAT): immediate and late results of a prospective multicenter randomized trial. AB - Mechanical rotational atherectomy with the Rotablator is widely used for percutaneous coronary revascularization, but the ideal debulking strategy remains unknown. The purpose of this study was to compare the immediate and late results after Rotablator using two treatment strategies: Large burrs (burr/artery ratio of >0.7) to achieve maximal debulking (lesion debulking strategy) or small burrs (burr/artery ratio < or = 0.7) to modify lesion compliance (lesion modification strategy). Two hundred twenty-two patients at six centers were prospectively enrolled in this study and randomly assigned to large (n = 104 patients with 118 lesions) or small (n = 118 patients with 136 lesions) burrs. The primary endpoint was final diameter stenosis at the end of the procedure, and secondary endpoints included inhospital angiographic and clinical complications, and target lesion revascularization at 6 months. Baseline demographic and angiographic characteristics were similar. There were no differences in procedural success, the extent of immediate lumen enlargement, inhospital ischemic complications, or late target vessel revascularization. However, compared with small burrs, patients randomized to large burrs were more likely to experience serious angiographic complications (5.1% vs. 12.7%, P < 0.05) immediately after atherectomy. This study suggests that a routine lesion modification strategy employing small burrs (burr/artery ratio < or = 0.7) achieves similar immediate lumen enlargement and late target vessel revascularization compared with a more aggressive debulking strategy (burr/artery ratio >0.7), but with fewer angiographic complications. PMID- 11387609 TI - Severe intracranial hemorrhage after emergency carotid stenting and abciximab administration for postoperative thrombosis. PMID- 11387610 TI - A unique pitfall in percutaneous coronary angioplasty of in-stent restenosis: guidewire passage out of the stent. AB - Three cases of in-stent restenosis are narrated, wherein, during balloon angioplasty of the lesion, the guidewire inadvertently exited out of the stent. The forward balloon progress was halted in this region. In the first case, the situation could only be realized when dilatation of a forcefully pushed small balloon avulsed the well-embedded stent. The mishap was averted in the subsequent two cases by reintroduction of a new guidewire. Some suggestions to avoid this eventuality are offered. Though the cases pertain to in-stent restenosis, the observations may be applicable to the procedures in general that entail passage of a guidewire through a stented area. PMID- 11387611 TI - Inadvertent "stent-wire bypass". PMID- 11387612 TI - Combined percutaneous biosense-guided laser myocardial revascularization and coronary intervention. AB - Laser myocardial revascularization is a promising new treatment strategy for patients with severe ischemic heart disease who are not candidates for conventional percutaneous or surgical revascularization. The open chest surgical approach to transmyocardial revascularization has been approved by the FDA for the treatment of angina in inoperable patients, but has had limited use as a stand-alone procedure. More recently, use of fiber-optic catheters has made it possible to use a holmium:yttrium aluminum garnet laser to perform percutaneous catheter-based transmyocardial revascularization. To the extent that many patients have a combination of ischemic sources, some amenable to conventional revascularization and some not, combination or hybrid approaches have been considered. We report herein two patients with class IV angina who underwent laser myocardial revascularization using the Biosense system and complex percutaneous coronary intervention during the same procedure. Areas amenable to conventional percutaneous coronary intervention (PCI) were so treated, and viable but ischemic areas were supplied by totally occluded native vessels and bypass grafts underwent Biosense-guided laser myocardial revascularization (LMR). As the results of more controlled and blinded studies of laser myocardial revascularization become available (if results continue to be promising) and a better understanding of the mechanism of action of this treatment modality is achieved, LMR-PTCA hybrid will be performed in increasing frequency. However, even after establishing LMR efficacy, studies of LMR-PTCA hybrid should be conducted to determine the efficacy of this approach. PMID- 11387613 TI - Guided laser myocardial revascularization with coronary angioplasty: the emperor shops for new clothes. PMID- 11387614 TI - Late stent thrombosis without antecedent brachytherapy: confirmation and treatment with rheolytic thrombectomy. AB - We describe two cases of late stent thrombosis (> 30 days postprocedure) following stenting without brachytherapy. Stent thrombosis was confirmed after successful treatment by catheter-based rheolytic thrombectomy. Predictors of stent thrombosis are reviewed and issues concerning the duration of combination antiplatelet therapy after stenting are discussed. PMID- 11387615 TI - Primary angioplasty for isolated right ventricular infarction. AB - We describe a case of isolated right ventricular infarction that has rarely been diagnosed antemortem. Electrocardiogram showed ST segment elevation in left precordial chest, right precordial chest, and inferior leads, which mimicked those of anterior and inferior left ventricular infarction. Coronary angiography revealed that culprit lesion was totally occluded right coronary artery. Infarcted artery was nondominant right coronary artery with branches supplying only right ventricular wall. Restoration of coronary blood flow was obtained by primary stenting and resulted in prompt ST segment normalization in all leads. Despite extensive right ventricular wall motion abnormality, subsequent right ventricular dysfunction was not observed. PMID- 11387616 TI - Endovascular repair of traumatic pseudoaneurysm by uncovered self-expandable stenting with or without transstent coiling of the aneurysm cavity. AB - Various surgical options for internal carotid or subclavian artery pseudoaneurysm repair have been reported; however, in general they have resulted in poor outcomes with high morbidity and mortality rates. Recently, these open surgical procedures have been partly replaced by percutaneous transluminal placement of endovascular devices. We evaluated the potential for using flexible self expanding uncovered stents with or without coiling to treat extracranial internal carotid, subclavian and other peripheral artery posttraumatic pseudoaneurysm. Three patients with posttraumatic pseudoaneurysm were treated by stent deployment and coiling (two cases) of the aneurysm cavity. In one case, a 5.0 x 47 mm Wallstent (Boston Scientific) was positioned to span the neck of the 9 x 5 mm size pseudoaneurysm (left internal carotid artery) and deployed. Angiography demonstrated complete occlusion of the pseudoaneurysm without coiling. In the second patient, a 5.0 x 31 mm Wallstent (Boston Scientific) was positioned to span the neck of the 9 x 7 mm size pseudoaneurysm (right internal carotid artery) and deployed. A total of six coils (Guglielmi Detachable Coils, Boston Scientific) were deployed into the pseudoaneurysm cavity until it was completely obliterated. In the third case, an 8.0 x 80 mm SMART (Cordis) stent was advanced over the wire, positioned to span the neck of the 10 x 7 mm size pseudoaneurysm of the left subclavian artery, and deployed. Fourteen 40 x 0.5 mm Trufill (Cordis) pushable coils were deployed into the pseudoaneurysm cavity until it was completely obliterated. At long-term follow-up (6-9 months), all patients were asymptomatic without flow into the aneurysm cavity by Duplex ultrasound. We conclude that uncovered endovascular flexible self-expanding stent placement with transstent coil embolization of the pseudoaneurysm cavity is a promising new technique to treat posttraumatic pseudoaneurysm vascular disease by minimally invasive methods, while preserving the patency of the vessel and side branches. PMID- 11387617 TI - Treatment of femoral artery pseudoaneurysm with percutaneous thrombin injection. AB - Pseudoaneurysm formation of the femoral artery is a well-known complication following catheter-based vascular procedures. Ultrasound-guided compression or surgical correction are commonly used for its repair. We describe a new method of treatment for femoral pseudoaneurysm. The pseudoaneurysm is visualized by contralateral angiography and thrombosed with a percutaneous thrombin injection while the distal vessel is isolated with a brief balloon inflation. Fluoroscopically guided percutaneous thrombin injection is a promising, minimally invasive technique for the treatment of iatrogenic pseudoaneurysm, especially in patients with compromised distal circulation. PMID- 11387618 TI - Intra-arterial thrombolysis (reteplase) and ReoPro for subacute thrombosis of the distal aorta accomplished by radial arterial access: the radial-kissing thrombolytic technique. AB - A 33-year-old woman with subacute thrombosis of the distal aorta after aorto-bi iliac stenting had local thrombolysis with reteplase in conjunction with systemic abciximab. The infusion was given as a bolus and then continuously for 14 hr by radial artery access with two selective kissing catheters. Patency of the stented segments was achieved with this technique in conjunction with resolution of her clinical symptoms. PMID- 11387619 TI - Transradial approach-from the evangelist's view. PMID- 11387620 TI - Evaluation of the percutaneous intramyocardial injection for local myocardial treatment. AB - Therapeutic angiogenesis requires the induction of new blood vessel formation for the treatment of peripheral vascular and coronary artery disease. Efficacious application of this new therapy requires optimizing multiple factors, including the therapeutic agent, dosing, frequency of administration, and delivery modality. In this study, a helical needle drug infusion catheter was applied for optimal application of percutaneous intramyocardial delivery (PIMD). (125)Iodine labeled albumin was injected by PIMD into the left ventricle myocardium in eight swine. After 1 hr, PIMD resulted in a high concentration of radiolabel at the treatment site; 16.4% +/- 2.1% of delivered and 81.4% +/- 2.6% of the total cardiac activity was concentrated at the site of delivery. The depth of needle penetration correlated with the myocardial retention of delivered protein. The myocardial retention of radiolabel in animals with shallow injections was 10.1% +/- 0.8%, compared to 18.9% +/- 3.3% retention after deep injections. The specific activity at the treatment site (radioactive counts per gram of tissue) was 115 +/- 36, 226 +/- 55, and 47 +/- 10 times higher compared to liver, lung, and kidney, respectively. Continuous coronary sinus and aortic blood sampling indicates that within 15 min following intramyocardial injection, a significant amount of nonretained protein is found within the coronary sinus. This study defines some of the parameters that can affect optimal application of PIMD and demonstrates that PIMD is a safe and efficient method for local drug delivery. PMID- 11387621 TI - In vivo assessment of stent recoil in normal porcine arteries: evaluation of contemporary stent designs. AB - Acute stent recoil has been observed following balloon deflation in normal and diseased coronary arteries, and the magnitude varies by stent design. We sought to evaluate acute stent recoil in five new stents. Twenty-five stents (four Crown, five Nir Conformer Royale, five Crossflex, five SupraG, and six GFX) were implanted in six Yorkshire pigs. All stents were expanded using a noncompliant balloon (balloon:artery ratio 1.2:1.0). Continuous ultrasound imaging was performed during stepwise balloon inflation and deflation using a 0.018" imaging core. Maximum cross-section areas (CSA) and minimal luminal diameter (MLD) were measured at 12 atm and immediately following balloon deflation. Maximum stent CSA matched expected balloon CSA. Area and diameter recoil were calculated as 1 - (CSAdeflation/CSAmax) and 1 - (MLDdeflation/MLDmax), respectively. Upon deflation, all stents showed recoil from maximal CSA. Area recoil was significantly lower for slotted-tube stents than modular stents (12.6% +/- 1.6% vs. 23.2 +/- 3.5%; P < 0.05). In compliant, nonatherosclerotic porcine coronary arteries, acute stent recoil for the four slotted-tube designs ranged from 8.4% to 18.0% by area. The modular stent tested was associated with significantly greater acute recoil than the slotted-tube stents. PMID- 11387622 TI - American College of Cardiology/Society for Cardiac Angiography and Interventions clinical expert consensus document on cardiac catheterization laboratory standards: summary of a report of the American College of Cardiology Task Force on clinical expert consensus documents. PMID- 11387625 TI - Forensic science: taking giant steps forward. PMID- 11387623 TI - Restenosis in one lesion in patients with multilesion stenting occurs even when the companion lesion is free of restenosis. PMID- 11387626 TI - DNA evidence will be admissible if the proper foundation is laid: advice for a forensic medicine expert. AB - The use and advance of DNA evidence in courts of law has mushroomed in the last few years to the point that it can almost be assumed that DNA evidence will be admitted if the proper foundation is laid. While a few basic principles are necessary for admissibility in all cases, each case stands on its own and depends upon its own fact situation and evidence to determine admissibility. The more advanced the science and specificity in each case, the easier for admissibility. To challenge admissibility, a lawyer has to challenge the foundation laid for the evidence. It only stands to reason that if you cannot attack the science, you have to attack the scientist, ie, the expert witness. This presentation is based upon the author's six years as a prosecutor and almost 23 years as a judge. It will give attendees some helpful advice on the purpose of an expert witness, how to be qualified as an expert witness, and some dos and don'ts, as well as hints as to how to become a better expert witness. PMID- 11387627 TI - Preservation and collection of biological evidence. AB - As the courts have placed greater emphasis on physical evidence during the past few decades, the initial stages of evidence examination have become increasingly important to the successful resolution of many criminal investigations. This emphasis on evidence collection and preservation is often manifested by many rigorous court challenges. This article reviews how the ability to introduce DNA test results in court is affected by methods used to recognize, document, collect, and preserve biological evidence. PMID- 11387628 TI - Application of low copy number DNA profiling. AB - Low copy number (LCN) DNA profiling is a technique sensitive enough to analyze just a few cells. When this kind of analysis is carried out, special considerations are needed to interpret the results. In particular, it is important to consider the implications of allele dropout and the possibility of contamination from a laboratory source. A rationale for interpreting LCN DNA is described. PMID- 11387629 TI - Short tandem repeat-based identification of individuals and parents. AB - Estimation of short tandem repeat (STR) multilocus genotype frequencies for the identification of individuals and estimation of allele frequencies for parentage assignment both depend on (a) testing a lot of loci, (b) high levels of polymorphism at each locus tested, and (c) independence among alleles. Independence is critical, because the estimation of multilocus genotype and gamete frequencies is based on multiplying individual allele frequencies to produce a composite frequency estimate. Independence among alleles at a locus is known as Hardy-Weinberg equilibrium, whereas allelic independence between loci is known as linkage equilibrium. The frequency at which individual identification may be declared is a matter of opinion, as there is no scientific way to specify certainty based on frequency estimates. Similarly absolute assignment of parentage is impossible in theory; in practice it is more difficult than individual identification, because only half as much information is available (gamete vs genotype frequency) and because mutation may confound parentage analysis. PMID- 11387630 TI - Comparison of PowerPlex 16, PowerPlex1.1/2.1, and ABI AmpfISTR Profiler Plus/COfiler for forensic use. AB - AIM: Several amplification and detection formats for the analysis of short tandem repeat loci are readily available to the forensic laboratory. Careful consideration must be given to the throughput, sensitivity, concordance, data interpretation, facility requirements, and costs of operation. The Pennsylvania State Police DNA Laboratory sought to establish that of any of the amplification or detection formats generally used in the United States generates concordant results and that the use of several formats within one laboratory provides a solution to the interpretation of difficult evidentiary samples. METHODS: Validation work consisting of sensitivity, precision, mixture, and substrate studies was performed by use of each of three detection formats (ABI Prism(r)310 Genetic Analyzer, ABI Prism(r)377 DNA Sequencer, and the Hitachi FMBIO(r)II Fluorescent Scanner) and three amplification systems (GenePrint(r) PowerPlex 16, GenePrint(r) PowerPlex 1.1/2.1, and AmpflSTR ProfilerPlus/COfiler). The results generated in each of the formats were compared, along with the problems incurred. RESULTS: All allele calls were concordant, with the exception of primer region variants, and all detection systems were sensitive and reliable. Even with the use of multiple formats, a general protocol can be written with only one set of interpretation guidelines. CONCLUSION: National databases can be used with input data from any of these formats. The use of several detection formats allowed the forensic scientist to select a system, based on sample quality, quantity, and throughput requirements. Interpretation issues resulting from complex mixtures, degraded samples, rare microvariants, internal primer variants, unusual heterozygote ratios, above or below ladder alleles, and potential tri-alleles can be verified. PMID- 11387631 TI - Interpretation of complex forensic DNA mixtures. AB - Forensic evidentiary samples routinely contain DNA from multiple contributors. The interpretation of these mixtures can be a challenging task for the DNA scientist. Several approaches are discussed (no calculation- qualitative statement; probability of exclusion; likelihood ratio estimates; presumptive genotype assignment based on peak heights), which have been employed to assess the significance of an inclusion/match when DNA mixtures have been detected in casework samples. These statistical approaches are discussed in light of technical challenges that can arise when evaluating evidentiary samples. PMID- 11387632 TI - Implementation of forensic DNA analysis on casework evidence at the Palm Beach County Sheriff's Office Crime Laboratory: historical perspective. AB - Palm Beach County is the largest of the 64 counties in the state of Florida, USA, with most of the area uninhabited and the population concentrated near the coastal region. The Serology/DNA Section of the Palm Beach County Sheriff's Office (PBSO) Crime Laboratory serves a community of approximately one million residents, and an additional million tourists visit Palm Beach County every year. In addition to the unincorporated county regions, there are thirty-four city police agencies, the Florida State Highway Patrol, several university security agencies, the local Federal Bureau of Investigation, and the county Medical Examiners Office that all use the PBSO Serology/DNA Laboratory for the analysis of casework evidence. The purpose of this manuscript is to provide laboratories that are in the process of initiating DNA analysis on casework with practical information regarding the decision-making processes that occurred during the development of the DNA testing program at PBSO. Many of the concerns addressed in the early 1990's are still a guide to the development of a quality forensic DNA analysis program in the year 2001. Issues, such as personnel, laboratory space, internal standard operating procedures, implementation of DNA analysis on casework evidence, and building a relationship with law enforcement personnel are discussed. PMID- 11387633 TI - Automation and high through-put for a DNA database laboratory: development of a laboratory information management system. AB - Automation and high through-put production of DNA profiles has become a necessity in every DNA database unit. In our laboratory we developed a Laboratory Information Management System (LIMS) controlled workflow architecture, which comprises a robotic DNA extraction- and pipetting-system and a capillary electrophoresis unit. This allows a through-put of 4,000 samples per person per year. Improved sample handling and data management, full sample- and batch histories, and software-aided supervision of result data, with a consequent average turn-around time of 8 days, are the main features of our new system. PMID- 11387634 TI - Establishing a large DNA data bank using the PowerPlex 1.1 and 2.1 systems. AB - In the early 1990's, the importance of establishing a DNA Data Bank of convicted sex offender samples for comparison to unsolved cases became apparent to the Virginia Division of Forensic Science to help identify potential perpetrators. Ultimately, through the expansion of the data basing law to include all convicted offenders and juveniles convicted of a crime that would be considered a felony if tried as an adult, the Division has successfully used the DNA Data Bank to aid the law enforcement community in solving crimes where the victim was unable to identify the perpetrator. As the number of offender sample analyses has increased, in combination with the number of analyses of cases where a suspect could not be identified, the number of DNA Data Bank hits has also significantly increased. Initially, in 1997, when the Division converted its DNA Data Bank program from the restriction fragment length polymorphism technology to the short tandem repeat technology, one offender hit occurred on average for every 2,900 convicted offender samples that were entered into the Data Bank. However, by December 31, 2000, one DNA Data Bank hit occurred on average for every 700 samples entered into the Data Bank. PMID- 11387636 TI - Identification of missing persons: the Spanish "Phoenix" program. AB - In 1999, Spain was the first country to officially start a National Program to try to identify cadavers and human remains which could not be identified by the use of traditional forensic approaches. This attempt is called "Phoenix Program". Two independent mitochondrial DNA (mtDNA) databases were generated, which can automatically compare and match identical or similar sequences. One is the Reference Database, with mtDNA sequences from maternal relatives of missing persons, who provide the samples (buccal swabs) voluntarily; the other is the Questioned Database, comprised of mtDNA data of unknown remains and cadavers. Although the first phase of the program (typing of all unidentified human remains) will probably not be completed until December 2003, positive identifications are being made in the interim. To date, more than 1,200 families have contacted Phoenix, and at least 280 reference samples and 48 questioned evidences have been analyzed. When mtDNA matches are found, another independent analysis is performed as a part of the quality control mechanism. Once a match is confirmed (so far in 6 cases), an attempt is made to analyze short tandem repeat (STR) loci. We call for international collaboration to make this effort valuable worldwide. PMID- 11387635 TI - DNA typing from skeletal remains: evaluation of multiplex and megaplex STR systems on DNA isolated from bone and teeth samples. AB - AIM: To evaluate the performance of three multiplex short tandem repeat (STR) systems (AmpflSTR Profiler, AmpflSTR Profiler Plus, and AmpflSTR COfiler), and a megaplex STR system (PowerPlex 16) on DNA extracted from the skeletal remains. By performing a microbial DNA challenge study, we also evaluated the influence of microbial DNA on human DNA typing. METHODS: A subset of 86 DNA extracts isolated from 8-50 years old bone and teeth samples, corresponding to 20 identification cases from mass graves in Croatia and Bosnia and Herzegovina, and to 4 paternity cases involving deceased parents in Spain, were analyzed by the above systems. RESULTS: Bone samples with no detectable human DNA (tested with Quantiblot), as well as teeth samples with detectable human DNA, were successfully amplified. Surprisingly, even in highly degraded samples, PowerPlex 16 offered very robust amplification for the both Penta E and Penta D markers. We observed a few non specific extra peaks of 202 and 308 base pairs, which appeared to match 16S rRNA of the Pseudomonas halodenitrificans. CONCLUSION: AmpflSTR Profiler Kit, AmpflSTR Profiler Plus Kit, the AmpflSTR COfiler Kit, and the PowerPlex 16 system are very sensitive multiplex STR amplification systems, which can be successfully used to obtain a multilocus STR profile from old teeth and bone samples with minimal amounts (pg) of human DNA or even with no detectable human DNA. PMID- 11387637 TI - Mass identification of persons missing from the break-up of the former Yugoslavia: structure, function, and role of the International Commission on Missing Persons. AB - The staff of the International Commission on Missing Persons (ICMP) is attempting to undertake the largest mass human identification effort in history. Through the generosity of numerous governmental and private corporations the ICMP has established or is currently establishing a strong network of political allies, family outreach centers, and DNA laboratories throughout the former Yugoslavia. Furthermore, the ICMP is currently working to streamline current technology as well as employ new technology in its efforts to assist in identifying missing individuals. ICMP will continue to act as a link between the family associations in the region and will synchronize the work of the DNA identification process in the countries affected by the war in the regions of the former Yugoslavia. In the longer term, ICMP seeks to contribute to the closure of the missing persons issue, to raise awareness of the human dimension of the missing persons tragedy, and to preserve a shared and common memory of the missing in the former Yugoslavia. PMID- 11387638 TI - Case work guidelines and interpretation of short tandem repeat complex mixture analysis. AB - Interpretation guidelines for short tandem repeat casework analysis are difficult to construct. As soon as a set of guidelines are developed, a new case evolves that does not fit the painstakingly written document. The casework analysts gather and amend the guidelines again, and again. This article seeks to demonstrate that general guidelines can be set and written such that it can be used for any detection format. Guidelines published by the Scientific Working Group for DNA Analysis Methods, a working group of DNA forensic experts in the United States, are used to set the format for the written protocol on interpretations. The rule "the interpretation of results in casework is a matter of professional judgment and expertise. Not every situation can or should be covered by a preset rule" is stressed. Development of minimum and maximum threshold values, heterozygote ratios, stochastic limits, and determination of major and minor components based on validation studies is discussed. The paper travels through setting criteria to evaluate internal lane standards and amplification controls. It continues with establishing ranges for interpretation and defining true alleles versus anomalies. Examples of a variety of profiles are given and the potential interpretation, using signal intensities and genetics. In addition, report writing strategies and wording routinely used by the Pennsylvania State Police DNA Laboratory System are given. PMID- 11387639 TI - Operating and managing a statewide DNA program. AB - The success of the Virginia Division of Forensic Science's DNA program is based upon managing the DNA program from a central location, consistency in the methods and procedures used, cooperation and continual dedication of the laboratory staff, as well as overseeing of all technical issues by a single technical leader. To streamline the analysis of the evidence samples, the most probative items of evidence are examined first for biological materials. Once biological materials are identified, DNA analysis is performed and the resulting foreign short tandem repeat profile is entered into the Combined DNA Index Systems for routine searching of the profile against Virginia's and the nation's convicted offender DNA Data Banks. In addition, the foreign profile is also searched against other foreign profiles obtained from previously analyzed evidence samples, thereby helping the law enforcement agencies in Virginia solve crimes that would otherwise go unsolved. PMID- 11387640 TI - When autosomal short tandem repeats fail: optimized primer and reaction design for Y-chromosome short tandem repeat analysis in forensic casework. AB - Y-chromosomal short tandem repeats (Y-STRs) are useful forensic DNA markers in investigation of sexual assault cases when a mixture of male and female DNA (e.g., in vaginal swabs) is present in a sample, especially when DNA of the male contributor is present only in very small amount compared to the DNA of the female victim. With autosomal STR analysis of male and female DNA, male DNA in mixtures can usually be detected and correctly interpreted only when it exceeds 5%. However, the amplification of some Y-STRs is known to result in polymerase chain reaction (PCR) products that are not associated with the Y-chromosome, but derive from the X-chromosome and/or autosomal regions. This can cause problems in the interpretation of results, particularly when female DNA is present in excess. Consequently, more specific and sensitive Y-STR primers and PCR conditions are needed. This paper presents two casework examples in which sensitive Y-STR multiplexes (with the addition of PCR enhancer) were successfully used in the analysis of mixtures of male and female DNA, the male component not interpretable by standard autosomal STR typing. PMID- 11387641 TI - Y chromosome-specific short tandem repeats in forensic casework. AB - Several case examples are presented to illustrate the usefulness of Y chromosome specific human DNA markers in a forensic setting. The markers used are the tetrameric short tandem repeats (STR's) DYS19, DYS389I, DYS389II, and DYS390. The main advantage of the Y-STR approach is the ability to detect the male component in a mixture of male and female DNA. It is also useful for the determination of the number of semen donors for mixtures of two or more male individuals. PMID- 11387642 TI - Application of Y-chromosomal STR haplotypes to forensic genetics. AB - This paper delivers population genetic data on Y-chromosomal short tandem repeat (STR) polymorphisms along with reports of unusual observations and casework. Population studies were carried out on the Y-specific STR polymorphisms DYS19, DYS385 I+II, DYS389 I+II, DYS390, DYS391, DYS392, and DYS393 in population samples from North India, Turkey, and Germany. In all three populations the vast majority of haplotypes was observed only once, especially in the Turkish group. Highly unusual cases are reported. In a German individual, we observed the variant allele DYS392*11.1, whereas a Turkish haplotype revealed a duplication at locus DYS19. Application of Y-chromosomal STR markers to forensic genetics was demonstrated in two cases: 1) a deficient paternity case, and 2) a father/son pair, where the Amelogenin primers failed to amplify the Y-homolog. In forensic genetics, Y-chromosomal STR polymorphisms are highly welcomed as an additional tool. PMID- 11387643 TI - Forensic mitochondrial DNA analysis: two years of commercial casework experience in the United States. AB - AIM: To describe mitochondrial DNA (mtDNA) forensic casework experience in a commercial laboratory in the United States. METHODS: Frequency statistics were kept for two years on all aspects of mtDNA forensic cases, including types of clientele, types of samples, levels of sample success and failure, site heteroplasmy, length heteroplasmy, contamination, rates of failures to exclude, and match statistics using a mtDNA sequence database. RESULTS: Low sample failure rate was observed, especially since an "ancient DNA" approach was used for samples with degraded DNA. Levels of contamination were low, and the observed site and length heteroplasmy did not confound the interpretation of results. The data collected from mtDNA haplotypes developed in casework showed extremely high diversity of haplotypes consistent with other formally developed databases. CONCLUSIONS: MtDNA forensic analysis in the private sector was successfully applied to many different types of samples overall, with minimal rates of complication due to sample handling challenges (degraded DNA, minimal samples, contamination) and sequence-specific phenomena (site and length heteroplasmy). PMID- 11387644 TI - Increasing the forensic discrimination of mitochondrial DNA testing through analysis of the entire mitochondrial DNA genome. AB - The principal limitation in forensic mitochondrial DNA (mtDNA) testing is the low power of discrimination that is obtained when common "mtDNA types" are involved in a case. Currently, an "mtDNA type" refers to the sequence within hypervariable regions I and II (HV1/HV2) of the control region, approximately 610 bp. In Caucasians, the most common HV1/HV2 type is found in approximately 7% of the population and there are 12 additional types found at greater than approximately 0.5% (ignoring HV2 C-stretch polymorphism). We are performing large scale sequencing of the entire mtDNA genome (mtGenome), approximately 16,569 bp, of individuals who have common HV1/HV2 types. Of 31 individuals with the most common HV1/HV2 type, only 3 still match after mtGenome sequencing. Similar high discrimination is seen in other common HV1/HV2 types. The sites that discriminate the various common HV1/HV2 types are generally not those that are known to vary widely in more diverse population samples. This indicates that complete mtGenome sequencing of selected HV1/HV2 types may stand as the best way for identifying maximally useful single nucleotide polymorphism sites outside of the control region. Our strategy for identifying SNP sites is useful in resolving U.S. Caucasian, Hispanic, and African American mtDNAs is discussed. We also discuss the development of homogeneous fluorogenic polymerase chain reaction assays that target phenotypically neutral sites for practical use in casework. PMID- 11387645 TI - Pentanucleotide short tandem repeat locus DXYS156 displays different patterns of variations in human populations. AB - AIM: To establish a database for the pentameric short tandem repeat locus DXYS156 from worldwide populations for routine genotyping in forensic identity testing and evolutionary biology. METHODS: Using polymerase chain reaction with a newly designed primer pair, we analyzed 1,408 male and female samples from 28 populations representing four major geographic groups. RESULTS: We observed 11 different alleles, which we sequenced and used to construct an allelic ladder. CONCLUSION: DXYS156 displays a contrasting pattern of X-linked and Y-linked variation among geographic regions, and between X and Y chromosomes. This complex allele distribution may be forensically useful for the ethnic differentiation of unknown stains. PMID- 11387646 TI - A compendium of human mitochondrial DNA control region: development of an international standard forensic database. AB - A compendium of human mitochondrial DNA (mtDNA) control region types has been constructed. This updated compilation indexes over 10,000 population-specific mtDNA nucleotide sequences in a standardized format. The sequences represent mtDNA types from the Scientific Working Group on DNA Analysis Methods (SWGDAM) mtDNA database and from the public literature. The SWGDAM data are considered to be of higher quality than the public data, particularly for counting the number of times a particular haplotype has been observed. PMID- 11387647 TI - Population variation of human mitochondrial DNA hypervariable regions I and II in 105 Croatian individuals demonstrated by immobilized sequence-specific oligonucleotide probe analysis. AB - AIM: To detect sequence variation in 105 Croatian individuals by the use of duplex polymerase chain reaction amplification of full-length hypervariable region I and II (HVI/HVII) products and subsequent hybridization to a linear array of 27 immobilized sequence-specific oligonucleotide (SSO) probes, which targets six regions within HVI and HVII, and two additional sites, 189 and 16093. METHODS: Chelex-extracted bloodstains were used for amplification of HV regions. In all cases, a single robust amplification was sufficient for immobilized SSO probe typing and subsequent direct sequence analysis for both HVI and HVII. This method, suitable for a range of forensic samples (including shaft portions of single hairs), was also applied to the analysis of 18 skeletal elements recovered from a mass grave. Using a panel of immobilized SSO probes, we have developed a rapid screening approach to mitochondrial DNA (mtDNA) haplotyping before direct sequence analysis. RESULTS: We established a reference sequence database of mtDNA haplotypes for 105 randomly selected Croatian individuals. Fifty different mitotypes were observed (33 unique). The most frequent mitotypes occurred 18 times or approximately 17.1% [111111 189 (A) 16093 (T)] and 11 times or approximately 10.5% [131111 189 (A) 16093 (T)]; all other mitotypes occurred 5% or less. The corresponding genetic diversity value for this database was approximately 0.952. The usefulness of establishing an mtDNA reference database with immobilized SSO probe testing has been demonstrated by determining the strength of a match comparison obtained for one skeletal element and a corresponding maternal reference from 18 specimens recovered from a mass grave. CONCLUSION: The sequence variation detected by the panel of immobilized SSO probes is sufficiently diverse to be used for identification of human skeletal remains from mass graves. The immobilized SSO typing strip targets polymorphic regions within HVI and HVII and is a useful identification tool for mass grave and mass disaster analysis, as well as for criminal casework testing. PMID- 11387648 TI - Development of a human DNA quantitation system. AB - The AluQuant Human DNA Quantitation System has been developed for human-specific quantitation of forensic samples. This system uses probes specific to repetitive genetic elements allowing quantitation without target amplification. Target immobilization is unnecessary with employment of solution hybridization. The AluQuant Human DNA Quantitation System uses a series of enzymatic reactions to produce a luminescent signal proportional to the quantity of human DNA present. This report demonstrates a range of quantitation from 0.1-50 ng of human DNA. Signal from non-human DNAs tested was insignificant and addition of non-human DNAs into a human sample did not alter quantitation. Lastly, the system was unaffected by degradation of sample through sonication. The AluQuant Human DNA Quantitation System is a simple and sensitive method for quantitating the concentration of human DNA in forensic samples. PMID- 11387649 TI - The Green Revolution: botanical contributions to forensics and drug enforcement. AB - Forensic botany encompasses many sub-disciplines, including plant anatomy, plant ecology, plant systematics, plant molecular biology, palynology, and limnology. Although the field of forensic botany has been recognized since the mid-1900's, the use of trace plant material as physical evidence in criminal casework is still novel. A review of published forensic casework that used plant evidence is presented here. Cases include the analysis of wood evidence in the Charles Lindbergh baby kidnapping, the use of pollen in establishing the location of a sexual assault, and pollen analysis to determine the time of year for burial in a mass grave. Additional cases discuss the use of plant growth rates to determine the time of a body deposit in a field, the use of diatoms to link individuals to a crime scene, and plant DNA typing to match seedpods to a tree under which a body was discovered. New DNA methods in development for plant species identification and individualization for forensic applications are also discussed. These DNA methods may be useful for linking an individual to a crime scene or physical evidence to a geographic location, or tracking marijuana distribution patterns. PMID- 11387650 TI - The singers. PMID- 11387651 TI - [Definition of a new temporal voice onset measurement]. AB - The aim of this study was to define a new temporal measurement of voice onset. If in fact it has been established that voice onset contains perceptive information which the listening panel uses to evaluate dysphonia, up till now there has been no objective measurement of this. In order to be able to use instrumental measurement of voice onset, it was first necessary to arrivee at a definition of the limits of onset and end. In electronics, the method of "10%-90%" is used to calculate the rise time (onset) of any signal. It is possible to apply this method to the voice signal using the intensity curve. We have made such measurements on 240 examples of the sustained lal sound produced by 60 dysphonic subjects and 20 normals. The results have shown that measurement of the duration of onset was in the main relevant for the mild (G1) and moderate (G2) dysphonias. We have concluded that the "10%-90%" method allows definition of the contours of onset, and measurement of its duration in a reproducible and standardised way. PMID- 11387652 TI - [Visualization of the vibratory movements of the vocal cords under asymmetrical conditions]. AB - In this study, we present a new system for measuring the vibration signal of the larynx which allows the vibration of each vocal cord in an excised larynx to be studied using a method of distant optical measurement. The set-up consisted of an insufflator and a device for immobilising the larynx, together with microprobes capable of creating mechanical pressure at the laryngeal ventricles. The whole system was computer-controlled. The measurements were carried out under normal and asymmetrical conditions on seven excised pig larynges. Measurement of the individual vibration of each vocal cord showed that the transient period of commencement of vibration (vocal onset) is influenced to a significant extent by the degree of asymmetry. Under conditions of extreme asymmetry, movement of the cords appeared to be aberrant, but not random because of the coupling mechanisms which cause "anchoring" of the two cords during the phase of closure. Thus one cord may vibrate at twice the frequency of the other. The method described has given us a better understanding of the mechanism of laryngeal vibration, and one which is in conflict with the theoretical harmonic model in many details. PMID- 11387653 TI - [Clinical significance of calculating the coefficients of Lyapunov in the objective assessment of dysphonia]. AB - Measurement of the vibration instability by means of jitter in cases of dysphonia fails to take into account certain features of the vocal signal, such as modulation or bitonatily. These abnormalities appear in jitter measurements as instability, without allowing any differentiation from random instability (to give 'true' instability). The authors suggest the use of the coefficient of Lyapunov, a measurement which is used for nonlinear dynamics. The vocal product of 179 normal and dysphonic subjects has been analysed. The coefficient of Lyapunov and the jitter were measured during the stable period of a sustained 'a' vowel. For the coefficient of Lyapunov, the authors used the algorythm which has been published previously [Giovanni et al, J. Voice 1999;13:341-354]. The measurements were compared with the perceptive analysis of the reading voice by a panel, and divided into four grade (G0 to G3). The coefficient of Lyapunov appeared to be more relevant than the jitter in distinguishing the various perceived grades. Precise recognition of the grade was obtained in 46% using the coefficient of Lyapunov as against only 36% using jitter. The addition of this new index to the existing puriparametric methods of voice analysis seems to be promising. PMID- 11387654 TI - Assessing efficacy of voice treatments: a guideline. AB - The proposal of this guideline or basic protocol is an attempt to reach better agreement and uniformity concerning the methodology for functional assessment of pathological voices. The purpose is to allow relevant comparisons with the literature when presenting/publishing the results of voice treatment, e.g. a phonosurgical technique, or a new/improved instrument or procedure for investigating the pathological voice. Meta-analyses of results of voice treatments are generally limited--and even impossible--due to the major diversity in assessing functional outcomes. A minimal, multidimensional set of basic measurements is proposed, suitable for all "common" dysphonias: it includes 5 different approaches: perception (grade, roughness, breathiness), videostroboscopy (closure, regularity, mucosal wave and symmetry), acoustics (jitter, shimmer, Fo-range and softest intensity), aerodynamics (phonation quotient), and self rating by the patient. The protocol is elaborated on the base of an exhaustive review of the literature, the experience of the Committee members, and of plenary discussions within the European Laryngological Society. Instrumentation is kept to a minimum, but considered essential for professionals performing phonosurgery. PMID- 11387655 TI - Clinical implementation of a multidimensional basic protocol for assessing functional results of voice therapy. A preliminary study. AB - 45 patients with various kinds of organic benignant voice pathology (vocal fold nodules, polyp, Reinke's oedema, unilateral paralysis, sulcus/scar) and/or with muscle tension disorders, for which a (pre-operative) functional voice therapy was indicated, were evaluated before and after this short period of voice therapy (4-26 sessions), in order to check a basic multidimensional protocol for assessing functional results of voice treatments, developed by the Committee on Phoniatrics of the ELS. Perception: Grade, Breathiness and Roughness from the GRBAS-scale, as rated by two experienced judges (0 = normal or absent; 3 = severely deviant or present). Acoustics: Jitter % and Shimmer % as computed by the MDVP program of Kay Elemetrics on a /a:/, at comfortable loudness and pitch. Also Fo-range and softest possible intensity were registered. Videostroboscopy: Glottal closure, regularity, mucosal wave and symmetry, as rated separately on visual analog scales of 10 cm by 2 experienced laryngologists. Phonation quotient: computed by dividing the vital capacity (ml) by the maximum phonation time (s) (best value of 2 x 3 trials). Self rating by the patient: voice quality in itself and general social/occupational handicap due to the voice problem were rated on 2 visual analog scales. Results show that there is a large variation in the interindividual and interdimensional results of the voice therapy: in a same patient, one dimension may be significantly improved while another one is significantly worsened. The 5 considered dimensions may be considered as low redundant. PMID- 11387656 TI - [Aspects of voice quality quantification]. AB - The present work is directed towards providing a complement to the current use of sustained sounds as the basis for the physical measurement of voice quality parameters. For many clinical observations, there is a clear lack of correspondence between the measurement results, for example of shimmer, jitter and harmonic to noise ratio, and the perceived quality of the speaking voice. Our work is motivated by the need to overcome this familiar problem. It has four main objectives: to use running speech as the basis for one set of measurements in the clinical protocol; to link physical measures based on the use of this data with both production and perception; to employ voice production data as the foundation for auditorily significant criteria; the presentation of quantitative comparisons using sustained sound and running speech data, derived for a range of pathological voice conditions, within the same clinical session. Our pilot observations are based on the use of clinical acoustic and laryngographic data from patients of 4 different clinical centres. PMID- 11387657 TI - [Forcing the voice and variance of speed: correlation between the speed of displacement of the center of gravity and the work of the postural muscles]. AB - Using stabilometry, our experimental protocol was designed to demonstrate a quantitative parameter of posture to study forcing of the voice. Analysis of the graphs has shown a perfect correlation between increases in variance of the speed of displacement of the centre of gravity and the increase in the electrical signal from the postural muscles, which is evidence their activation during forcing of the voice. As well as the clinical applications of this protocol, it seems necessary to undertake other studies such cinematography recording of respiration. PMID- 11387658 TI - [A comparative acoustic study of the speaking and singing voice during the adolescent's break of the voice]. AB - The observation of the vocal evolution of adolescent singers has shown it takes place in two stages, the singing voice changing after the speaking voice. The same pattern has been encountered and made more explicit with a study of 50 non singer adolescents. It thus appears that the average pitch of the speaking voice deepening by one octave is not by itself the sign that the break of the voice has ended. This study also shows the individual nature of adolescent vocal evolution and its length (up to two years in one out of four cases). PMID- 11387659 TI - Notes on voice and speech disorders in ancient and Byzantine Greece. AB - Voice and speech were considered important by the Ancient and Byzantine Greeks and their disorders were examined and treated on a reasoned basis, without prejudice, by such famous doctors and practitioners of these periods as Hippocrates, Aristotle, Galen, Orebassius and Paul the Aeginitian. In this article, we present the voice and speech disorders which were described and analysed in the texts of Ancient and Byzantine Greek doctors and historians. PMID- 11387660 TI - [Functional results of velopharyngoplasty: apropos of 55 cases]. AB - From 1991 until 1998, 55 three to twelve year-old patients underwent a velopharyngoplasty to correct velopharyngeal insufficiency. They had indeed severe hypernasality with or without gross reflux of food matter into the nasal cavities or behavioural disturbances. Eighty-two percent had a closed cleft palate. Forty percent presented with mental retardation, heart diseases or multiple syndromal defects. They all had had a previous speech therapy for a long (months) or a very long (years) period of time. In the post operative period, hypernasality disappeared totally or partially in eighty-five percent; reflux disappeared in almost all cases. Middle ear pathologies were not more frequent and were also less severe. Behavioral disturbances associated with a severe speech defect were also less pronounced. PMID- 11387661 TI - Personality study in profoundly deaf adults. AB - The difference with which the pre-lingual and post-lingual profoundly deaf confront their deafness is evident: the reticence of many pre-lingual deaf persons when faced with technological advances and genetic investigations is not present among the post-lingual deaf. This article looks at this difference from the person as a whole, and aims to carry out a comparative study of the personalities of both groups and of the non-deaf. To this end, three samples were taken (one of pre-lingual deaf persons, one of post-lingual deaf persons and a third of non-deaf persons) and the Mini-Mult personality questionnaire was employed as a measuring instrument. Significant differences were found in 5 clinical scales: Hs (hypochondria), Hy (hysteria), Pa (paranoia), Pt (psychastenia), Sc (schizophrenia). The differences between the deaf and the non deaf are statistically significant. The profile obtained for the pre-lingual deaf is high on the schizophrenia scale (Sc), and suggests a kind of person where the said scale might detect isolation derived from deafness. The profile of the post lingual deaf, marked by the hypochondria scale, shows persons with severe hypersensibility and fears regarding their state of health. The significant influence of variables such as level of education and state of employment on the post-lingual deaf and, in particular, on his feeling of loneliness and his worries regarding his deafness, his state of mind and his possible paranoiac tendencies, makes them factors that must be taken into consideration when fundamentally evaluating the personality of a post-lingual deaf person. These personality traits influence the way a profoundly deaf person confronts his deafness. PMID- 11387662 TI - [Importance of the stapedial reflex in the diagnosis of several pathologies]. AB - The stapedius muscle plays an important role in auditory physiology. Its function may be assessed using a test named impedancemetry, where its reflex is measured. For the stapedial reflex to function, it is necessary that the middle ear and the afferent and efferent pathways are intact. Several pathologies of the middle and internal ear, as well as other otoneurological disorders and systemic illnesses, may change the physiology of the stapedius muscle. This review has incorporated various bibliographic sources with the objective to show the modifications of the stapedial reflex in several types of pathology. It reveals that the research of the stapedial reflex as well as its interpretation constitute a very important diagnostic supplement in the clinical audiology. PMID- 11387663 TI - [Pulmonary aging]. PMID- 11387664 TI - [Aspergillosis in birds]. PMID- 11387665 TI - [Biology and pathogenicity of fungi]. AB - Men live with a multitude of fungal species and few species can become parasites. Some species are obligatory pathogenic (dermatophytes, dimorphic fungi...) and the majority is opportunistic pathogen. Host-parasite complex is indissociable. For example, Candida albicans is a commensal of the gastrointestinal tract and can dramatically proliferate when host defences are alterated. Passage from the commensal status to pathogenicity is accompanied by modifications of the fungus, which then act as virulence factors (development of filamentous forms, changes in surface hydrophobicity...). Biologic, genetic and antigenic modifications are also observed. Colonisation and adhesion, penetration, multiplication and survival are necessary for infectiosity in a host. Host defence mechanisms are described with non-specific mechanisms by humoral and cellular factors and more specific immune mechanisms involving antibodies, T lymphocytes and the role of cytokines. PMID- 11387666 TI - [Laboratory diagnosis of visceral mycoses]. AB - Prognosis of visceral mycosis depends on an early and well-adapted treatment. So, diagnosis must be done as early as possible. The clinician must first think of mycosis in front of a fever resistant to antibiotics or an unusual symptomatology, particularly in patients at risk for this opportunistic infection. The laboratory role is then essential. Isolation of the responsible fungus must be the first step. It leads to confirmation of diagnosis and to the choice of the best antifungal treatment. The search of antibodies and, in immunocompromised patients, of circulating antigens, is of precious help when no fungus is isolated or to confirm its pathogenicity. Unfortunately, diagnosis is often difficult. New molecular biological techniques should improve the quality and delay of mycological diagnosis. PMID- 11387667 TI - [Visceral candidiasis]. AB - Invasive candidiasis is a frequent infection in compromised patient. Several risk factors have been identified and include neutropenia, broad-spectrum antibiotherapy, colonisation with a Candida spp. and presence of central venous catheter. Candidemia is the most common clinical aspect. Diagnosis is based on positive culture of blood, skin biopsy or fine needle aspiration of a deep-seated lesion. Serology is not helpful in severely immunocompromised patients. Prophylaxis is based on strict hygiene and, in neutropenic patients, oral fluconazole. Treatment of an invasive candidiasis depends on the localisation of the infection, of its acute or chronic evolution, on the species involved and on underlying condition. Amphotericin B deoxycholate or in lipid formulation and fluconazole are the antifungal drugs of choice. Removal of a central venous catheter should always be discussed in candidemia. PMID- 11387668 TI - [Invasive aspergillosis]. AB - Invasive aspergillosis remains a life-threatening complication in immunocompromised patients. The pulmonary involvement by the aspergillosis is the most common setting. The dissemination of the disease is correlated with the worse prognosis. The overall improvement of the prognosis needs a global diagnostic and therapeutic strategy. In neutropenic patients (mostly at risk of invasive aspergillosis), an early diagnosis can be achieved by systematic use of thoracic CT scan to search halo sign (that seems to be quite specific of the diagnosis in this setting). The early initiation of the antifungal treatment (conventional or liposomal amphotericin or new azole compounds) could be combined with a surgical approach if necessary. This approach could be able to improve the prognosis of aspergillosis. However, the outcome of these patients remains also correlated to the underlying disease. PMID- 11387669 TI - [Cryptococcosis]. AB - Cryptococcus neoformans, an encapsulated yeast is responsible for life threatening infection in patients with cellular immune defect especially those with AIDS. The most frequent presentation of cryptococcosis is a disseminated meningoencephalitis without acute onset. The diagnosis is based on direct examination with India ink staining, detection of soluble capsular polysaccharide in body fluids, culture of the micro-organism or histology. Looking for other localisations and for factors of poor prognosis is mandatory before selecting the right treatment regimen. In case of meningitis, one should prefer a combination of amphotericin B and 5 fluorocytosin, followed by a triazole, usually fluconazole until sterilisation of all infected sites is achieved or even for life-time maintenance therapy in case of sustained immune defect. PMID- 11387670 TI - [Emerging visceral mycoses]. AB - The emergence of new fungal pathogens such filamentous fungi (Scedosporium, Fusarium) or yeast (Trichosporon) is a real problem for doctors who treat immunocompromised patients. These fungi present in our environment (Scedosporium, Fusarium) can also be on the skin (Trichosporon). They are responsible for invasive infections cause of morbidity and mortality because patients are immunocompromised and the fungi are resistant to antifungal treatment. Clinical manifestations are seldom specific, but cutaneous localisations are frequent. They are either the portal of entry of the infection or a metastatic localisation. The role of laboratory for identification of the fungus is essential. PMID- 11387671 TI - [Endemic mycotic infections]. AB - AIDS epidemics and intercontinental travels in endemic areas have increased the incidence of endemic mycoses (histoplasmosis, coccidioidomycosis, blastomycosis, penicilliosis). Environmental dimorphic fungi, through an aerial contamination cause them. Frequent in the HIV patients living in endemic areas, they represent an AIDS definition criterion. Most of primary infections are asymptomatic, they may also present as influenza or pneumonia, that will spontaneously recover. A secondary dissemination may especially occur among immunocompromised hosts involving most often the skin, central nervous system and bones. Lastly, a chronic pulmonary presentation may also occur. Direct examination and histology, cultures and serologies establish diagnosis. In all cases of disseminated or chronic infections, a long-term treatment is necessary, using amphotericin B and azoles. Life-time secondary prophylaxis is recommended in AIDS patients. PMID- 11387672 TI - [Choice and use of antifungal drugs]. AB - During the last decade the incidence of deep mycotic infections has continued to increase dramatically. This new epidemiological feature is mostly due to the expanding population of at risk subjects submitted to intensive and protracted immunosupression. Amphotericin B has a broad spectrum and has remained the drug of choice for these life threatening invasive fungal infections. However adverse events, particularly renal insufficiency, are limiting factors in achieving an effective dose. Since the 1990s the number of oral and parenteral antifungal agents has significantly increased. The use of phospholipid carriers for amphotericin B or cyclodextrine for triazoles improved safety profil and/or pharmacokinetics. New families of drugs with a new target in the fungal cell are under clinical investigation and should be soon available. Comparative studies, consensus on diagnosis criteria and practice guidelines for the treatment of fungal infections will contribute to a better management of the patients and optimize the use of antifungal agents. PMID- 11387673 TI - [Visceral mycotic infections. In clinical practice]. PMID- 11387674 TI - [Acute rhino-sinusal infections in adults: etiology, diagnosis, treatment]. PMID- 11387675 TI - [Iatrogenic diseases linked to the use of drugs and common medical equipment]. PMID- 11387676 TI - [Nonsteroidal anti-inflammatory agents: principles and rules of use]. PMID- 11387677 TI - [Intestinal obstruction of the colon: physiopathology, etiology, diagnosis, treatment]. PMID- 11387678 TI - [H2 receptor antagonists and proton pump inhibitors: principles and rules of use]. PMID- 11387679 TI - [Hospital (public and private) and ambulatory care systems: structures and principles of charges]. PMID- 11387680 TI - [Urinary incontinence in adults: diagnostic trends]. PMID- 11387681 TI - [Intracytoplasmic sperm microinjection]. PMID- 11387682 TI - [Lawsuits against surgeons in XVIIth-century Lorraine]. PMID- 11387683 TI - [Anesthesia in France]. AB - Anaesthesiology is a recent medical speciality, established in 1965, that is formally termed in French Anesthesie-reanimation (Anaesthesiology-intensive care). Since its creation, the discipline has had a range of activity including anaesthesia, intensive care, prehospitalisation emergencies, acute and chronic pain and diverse follow-up. In France 8 million anaesthesias are performed per year, by 8,500 physicians specialised in anaesthesiology, assisted by 7,500 anaesthesiology nurses. It is estimated that each year 13.5% of the French population undergoes anaesthesia, with variations related to sex and age. Over 15 years, the number of anaesthesias performed increased 2.2 fold, due to the increase in certain types of surgery (especially orthopaedics) but also obstetrics (35 times more peridural anaesthesias) and the explosion of GI endoscopy (50 times increase). This increased activity involves patients at the extreme ranges of life, young women and fragile persons. Local and regional anaesthesias represent 21% of those performed, and outpatient anaesthesia is 12 times that of 15 years ago. PMID- 11387684 TI - [Judicial and contentious aspects of anesthesia]. AB - The practice of anaesthesia is surrounded by the same legal texts as in other medical fields and by a decree, which is specific to it. This decree notably prescribes a preliminary consultation by an anaesthetist, in a lapse of time allowing change to eventually the planned strategy or to optimise preoperative care. It is therefore important that all medical data useful to the patient assessment should be available at this time. Since the application of professional standards, claims related to the anaesthetic practice have decreased in the United States, especially because of the reduction in perioperative hypoxic injuries. It should be noted that the development of such standards aims first at improving patient's care. Claims reduction could be consider as a secondary consequence. Other means of improving our practice deserve to be developed, such as national confidential surveys. PMID- 11387685 TI - [General anesthesia]. AB - General anaesthesia is a reversible loss of consciousness induced and maintained with a hypnotic drug given either by venous injection and infusion, or by inhalation. A potent opioid is usually associated to inhibit the transmission of pain and thus to lessen sympathetic and endocrine reactions to nociceptive stimuli. Myorelaxation is used to facilitate tracheal intubation and surgery. Whatever the anaesthetic protocol use, the patient and anaesthesia machine require close monitoring. In addition to vital signs, the depth of anaesthesia may be monitored using automated electroencephalographic analysis and myorelaxation should always be monitored using a nerve stimulator, but pain or analgesia evaluation is only based on clinical signs of sympathetic stimulation. Because anaesthesia-related death and morbidity have decreased considerably, future improvements in outcome should concern perioperative comfort, i.e. prevention of cognitive disturbances, nausea, vomiting and pain. PMID- 11387686 TI - [Locoregional anesthesia]. AB - Local anaesthetics used for regional anaesthesia inhibit impulse depolarisation through blockade of sodium channels. Regional anaesthesia accounts for 20% of all anaesthetic procedures performed in France. Spinal anaesthesia is the most common technique used peroperatively, while epidural anaesthesia is used in labouring women and for postoperative pain control. Nerve stimulation allows a safer an easier approach to peripheral blocks. Regional anaesthesia is mainly indicated for limb or eye surgery, and for mildly invasive surgical procedures. Regional anaesthetic techniques are the most efficient to control postoperative pain and to facilitate patient rehabilitation. The risk related to the practice of regional anaesthesia compares with that of general anaesthesia and is thus very low. Main complications are epidural haematoma, cardiac arrest due to inadvertent intravascular injection of local anaesthestics, and peripheral nerve damage. Regional anaesthesia requires the same technical environment and careful monitoring as general anaesthesia. PMID- 11387687 TI - [Anesthesia risks]. AB - Both effects of anaesthesia and analgesia are involved in the anaesthesia risk. Most often, anaesthetic accidents appear to be related to hypoxia. Applying anaesthetic security principles allow controlling risk. Systematic use and check of monitoring instruments and anaesthesiology department organisation appear as major preventive measures. Independent operative risk is related to individual, surgical and anaesthetic factors, which interact with one another. Individual preoperative evaluation is one of the great challenges of anaesthesiology. The establishment of an aimed perioperative anaesthetic strategy is the best warrant of the operative risk reduction. Assessment of individual benefit to risk ratio takes also into account the experience of the surgeon and anaesthesiologist. It implicates the patient's choice as well his general practitioner as a privileged advice. PMID- 11387688 TI - [Anesthesia in coronary disease]. AB - Patients undergoing non-cardiac surgery and who have coronary artery disease run a high risk of myocardial infarct. Diagnosis and postoperative management is presently based on assessment of cardiac troponin I. Associated with optimal postoperatory management, several approaches can limit the occurrence of postoperatory coronary complications and improve the life expectancy of surgical patients with high cardiovascular risk. They include the possibility of preoperatory myocardial revascularisation and the prophylactic, postoperatory administration of cardiovascular medications (non-steroidal anti-inflammatory drugs, beta-blockers and alpha 2 receptor agonists). PMID- 11387690 TI - ["Pedagogic education" of teachers]. PMID- 11387689 TI - [Postoperative analgesia]. AB - Postoperative analgesia is not adequate in France. An improvement is necessary for both the patient's comfort and the possible reduction of postoperative morbidity. The cornerstone of postoperative analgesia is regular evaluation of pain allowing adapted treatment. The organisation of postoperative analgesia requires teaching of medical and paramedical staff, the agreement on protocols with designation of a pain referent according to a quality insurance program. Available treatments include non-opioid and opioid drugs and local anaesthetic agents. Main difficulties include interindividual variability, and analgesia on movement. Patient-controlled analgesia allows adaptation of opioid treatment to patient's needs. The combination of drugs and techniques may allow a better analgesia, and a reduction of its side effects. PMID- 11387691 TI - [Hemorrhage associated with delivery. Diagnosis, treatment principles]. PMID- 11387692 TI - [Tobacco addiction. Epidemiology and pathology associated with tobacco]. PMID- 11387693 TI - [Non-traumatic meningeal hemorrhage]. PMID- 11387694 TI - [Myocardial infarction. Epidemiology, physiopathology, diagnosis, etiology, complications, treatment]. PMID- 11387695 TI - [Antibacterial antibiotics. Classification, principles and rules of use]. PMID- 11387696 TI - [Ocular traumatic injuries. Etiology, diagnosis, treatment principles]. PMID- 11387697 TI - [Acute diarrhea in the adult. Diagnostic orientation and management in emergency situations]. PMID- 11387698 TI - [Hearing loss is often too long misdiagnosed. Are we too loud? (interview by Dr. Brigitte Moreano)]. PMID- 11387699 TI - [Periarthritis humeroscapularis. Radiotherapy for pain?]. PMID- 11387700 TI - [Diagnosis in back pain involves not only the body. Investigate psychological stress to determine origin]. AB - With a prevalence of up to 40%, back pain is one of the most common symptoms encountered in the doctor's office. Some 10% of the patients with severe pain become chronic cases. For the investigation of causal factors, the diagnostic work-up should cover not only somatic, but also (urgently) psychological, emotional and cognitive factors, as well as passive disease coping, depressive moods, and subjective attitudes to the condition. On the somatic side, overstraining of the muscles themselves, and lack of exercise play an important role. For the somatic work-up, the task is to search for pain triggers and secondary disrupting factors. Major clues are provided by the case history and an analysis of muscular status. For the identification of psychosocial factors, careful history-taking must be supplemented by simple psychological test instruments, such as Beck's depression inventory. PMID- 11387701 TI - [What helps in back pain? Guideline for symptomatic therapy]. AB - Both in acute and chronic, unspecific back pain, the myofascial pain syndrome resulting in muscular dysbalance is a major factor. For the differential diagnosis, however, consideration must always be given to concomitant symptoms (neurological deficits, general symptoms, signs of osteopathy). Pathophysiologically, the active trigger point corresponds to a contraction in the muscle fibers that forms in the region of the a neuromuscular endplate, and leads, via biochemical processes, to the stimulation of mesochymal nociceptors. Symptomatic treatment of acute and chronic back pain may be broken down into a) physical measures, b) local therapeutic regimens, c) systemic pharmacotherapy. As medication, non-steroidal antiinflammatory drugs, non-opioid analgesics, opioid analgesics, muscle relaxants and antidepressives are available, and are dose matched to the severity and stage of the condition. The spectrum of therapeutic options is outlined. PMID- 11387702 TI - [Comparison of muscle relaxants and spasmolytic drugs. Antispasmodics against pain]. PMID- 11387703 TI - [Diabetic foot syndrome. When is surgery reliable and necessary?]. PMID- 11387704 TI - [Emergencies in general practice, 13. Acute urinary retention]. PMID- 11387705 TI - [Disarming Lyme borreliosis. A chameleon among infections]. PMID- 11387706 TI - [Treatment of group A streptococcal tonsillopharyngitis. 5 days cephalosporin is as effective as 10 days penicillin]. PMID- 11387707 TI - [Diagnostic quiz. Hyperglycemia in congenital hemihypertrophy. Cushing syndrome due to bilateral adrenal gland adenomas in Klippel-Trenaunay syndrome]. PMID- 11387708 TI - [Nicotine-free pill. Easy smoking cessation]. PMID- 11387709 TI - [Antihypertensive drug. Dual action lowers blood pressure more effectively]. PMID- 11387710 TI - [Seasonal allergic rhinitis. Nasal obstruction can also beh improved]. PMID- 11387711 TI - [Parkinson disease. Value of COMT inhibitors is verified]. PMID- 11387712 TI - [Prof. Bernd Simon on nonsteroidal anti-inflammatory drugs. Gastrointestinal complications are unpredictable]. PMID- 11387713 TI - [Selective TNF-alpha inhibition in Crohn disease. Emergency help in problem cases]. PMID- 11387714 TI - [Drug therapy of dementia. Improving cognitive performance]. PMID- 11387715 TI - [18. Depression: A challenge for the general practitioner]. PMID- 11387716 TI - Does contraction of mesh following tension free hernioplasty effect testicular or femoral vessel blood flow? AB - Prosthetic mesh can contract by 20-75% of its original size within ten months after implantation. We set out to determine whether this contraction has any effect on testicular or femoral vessel blood flow following open or laparoscopic hernia repair. Twenty patients who underwent mesh repair of a primary unilateral inguinal hernia repair by Open (10) or Laparoscopic (10) methods a median of 3 years previously were investigated by ultrasound to determine the haemodynamic characteristics of the testis and femoral vessels. There was no significant difference in testicular blood flow, volume or echogenicity between the different types of repair or the contralateral side. The vertical and transverse dimensions of the femoral artery and vein were similar in all groups as was blood flow. Mesh contraction following inguinal hernioplasty does not adversely affect the testis or femoral vessels and can be used safely for both anterior and preperitoneal approaches. PMID- 11387717 TI - Determinant factors of pain after ambulatory inguinal herniorrhaphy: a multi variate analysis. AB - Pain is common after inguinal herniorrhaphy. The objective of our study was to evaluate the significance of various clinical factors on the level of post operative pain after ambulatory inguinal herniorrhaphy. Between January, 1996 and December, 1998, 239 ambulatory inguinal hernia repair patients were recruited. Operative techniques included nylon darn (n = 152), modified Bassini repair (n = 56), and prolene mesh hernioplasty (n = 30). Linear analogue pain scores--ranging in value from 0 to 10--were assessed by telephone interviews on the first and third post-operative days. Uni-variate and multi-variate analyses were performed to identify the significant independent determinant factors affecting the severity of post-operative pain. Clinical factors studied were age, sex, operative technique, hernia anatomy and post-operative complication(s). By uni variate analysis, patients of age < or = 50 years and indirect inguinal hernia were associated with a significantly higher pain score on the first postoperative day 1. On post-operative day 3, patients of age < or = 50 years, with an indirect inguinal hernia and modified Bassini repair reported a significantly higher pain score. Following inguinal herniorrhaphy, multiple regression analysis showed that age was the only independent predictive factor of pain score on post-operative days 1 and 3. In conclusion, post-operative pain was not affected by surgical technique, sex, hernia anatomy and post-operative morbidity. Only age had a significant influence on the post-operative pain score following ambulatory inguinal herniorrhaphy. Therefore, the age of a patient should be taken into consideration when prescribing post-operative analgesics. PMID- 11387718 TI - Bassini vs Lichtenstein: two basic techniques for inguinal hernia treatment. AB - Two techniques for inguinal hernia treatment are compared: the plastic repair of Bassini and that of Lichtenstein. The authors stress the differences between these two techniques by reporting and comparing the results obtained during four years of operating on 381 patients with the Bassini technique and 357 with the Lichtenstein technique. PMID- 11387719 TI - Traumatic diaphragmatic hernias: a report of 26 cases. AB - Traumatic diaphragmatic hernias, when diagnosed many years after the traumatic event, are observed in about 10% of diaphragmatic injuries. Due to coexisting injuries and the silent nature of diaphragmatic injuries, the diagnosis is easily missed or difficult. The medical records of 26 patients, who were treated for diaphragmatic hernias during the last 20 years, were analysed retrospectively. The patients were divided into acute phase and late-presenting groups, in whom emergency surgery and elective intervention were performed respectively. Chest radiography was diagnostic in 34.6% (n = 9) of patients. 92.3% of the hernias were on the left side, while the most common herniated organs were the stomach (31.8%) and the colon (27.2%). Coexisting injuries were recorded in 38.4% (n = 10) of the patients. Primary repair was predominantly used (92.3%). The hospitalisation period was longer in the late-presenting group (24.1 +/- 18.8 vs. 14.3 +/- 7.7 days). Two deaths occurred in the late-presenting group. Diaphragmatic hernia should be suspected in all blunt abdominal trauma patients. Prompt surgical repair is the treatment of choice in all traumatic diaphragmatic hernias. PMID- 11387720 TI - Prosthetic repair of incisional hernia in kidney transplant patients. A technique with onlay polypropylene mesh. AB - The employment of synthetic mesh for incisional hernia repair in kidney transplanted patients is rarely reported in the present literature. Many authors believe that mesh employment in such conditions is not safe due to fear of mesh related complications. From 1965 through 1999, a total of 1685 kidney transplants were performed at our Kidney Transplant Unit and 19 patients developed eventrations in the kidney transplant incision, an incidence of 1.1%. From September 1996 eight of these patients had prosthetic repair of the abdominal wall with onlay polypropylene mesh. All patients were under immunosuppressive therapy with prednisone, ciclosporine and azathioprine. Mean age was 48.8 years, mean body mass index was 22.5 and mean number of previous abdominal operations was 2.5. A large polypropylene mesh (Marlex mesh) was fixed over the aponeurosis after primary closure of the aponeurotic borders, as an onlay graft. There was neither morbidity nor mortality associated to the surgical procedure. No recurrences or long-term complications associated with mesh employment were verified after a follow-up ranging from one year to three years. We concluded that prosthetic repair of incisional hernia in transplanted patients can be performed routinely. PMID- 11387721 TI - Inguinal hernia repair in the Amsterdam region 1994-1996. AB - In the Netherlands, approximately 30,000 inguinal hernia repairs are performed yearly. At least 15% are for recurrence. New procedures are being introduced creating discussion on which technique is the best. Currently it is not possible to choose on evidence alone because of the long follow-up that is needed. In 1996 an inventory was taken of all inguinal hernia repairs that were performed in the Amsterdam region (9 hospitals). These results were compared with the results from a similar study performed in 1994. Major changes in treatment strategy were noted. The Bassini repair was replaced by Shouldice and Lichtenstein techniques. There was a significant increase in the use of prostheses for both primary and recurrent inguinal hernias. There was no significant decrease in the percentage of operations performed for recurrent hernia from 19.5% to 16.8%. However, there was a significant decrease in operations performed for early recurrences (5.1% 3.4%) (p = 0.05). These results suggest that the Shouldice and Lichtenstein repairs may be superior to the Bassini repair in terms of early hernia recurrence. PMID- 11387722 TI - Laparoscopic incisional and ventral herniorraphy: our initial 100 patients. AB - A review of our initial 100 patients upon whom we attempted a laparoscopic repair of either a ventral and incisional hernia is presented. The average follow-up period of these individuals was 51 months. The operation was completed with the laparoscopic technique in 96 cases. The average defect size was 155 cm2 and the average prosthetic biomaterial size to repair these defects was 214.8 cm2. The major complication rate was 4.1%. The incidence of recurrence in these patients was 9.3%. In all of these cases of recurrence, the method of attachment was that of staples or spiral tacks alone. In 5 patients, it appeared that the prosthesis was too small to cover the defect adequately. We believe that this is an effective operation but one that has two technical mandates. The prosthetic biomaterial (DualMesh) must cover the fascial edges by a minimum of a three centimeter overlap. Additionally, the attachment of the patch by staples or tacks alone is inadequate; consequently, the herniorraphy must include the use of through and through sutures to assure adequate fixation of the prosthesis. PMID- 11387723 TI - Inguinal endometriosis or irreducible hernia? A difficult preoperative diagnosis. AB - Two cases of endometriosis infiltrating the round ligament and associated with an inguinal hernia are presented. The initial diagnosis was irreducible hernia, since this rare association often causes unusual preoperative symptoms and diagnostic problems. Diagnosis is frequently made by histologic examination. Surgery is the treatment of choice both for hernia and for endometriosis, and is locally curative. However, in a fertile woman with a painful mass in the inguinal region the possibility of endometriosis should be considered, and if suspected at inguinal exploration a laparoscopy should be made to rule out the presence of intraperitoneal endometriosis. PMID- 11387724 TI - Does every hernia demand a mesh repair? A critical review. PMID- 11387725 TI - A strangulated Spigelian hernia mimicking diverticulitis. AB - Spigelian hernias are true interparietal hernias that are frequently not associated with a palpable mass and, as such, are not frequently diagnosed before surgery. Reported herein, is the serendipitous discovery of an incarcerated Spigelian hernia that was discovered on a CT scan being performed for presumed diverticulitis. PMID- 11387726 TI - Pre-vascular hernia: a rare cause of chronic obscure groin pain after inguinal hernia repair. AB - A rare case of pre-vascular hernia is reported in a woman complaining of chronic obscure groin pain following an inguinal hernia repair. The condition was only diagnosed by means of a herniogram, emphasising the value of this investigation in unexplained groin pain. The hernia was successfully repaired using a polypropylene mesh plug, a simple technique widely employed in both femoral and recurrent inguinal hernia, but never before described in pre-vascular hernia. PMID- 11387727 TI - Bioabsorbable membrane prevents adhesions to polypropylene in rats. PMID- 11387728 TI - On the extraperitoneal origin of hernia. PMID- 11387729 TI - Abdominal wall hernia repair in patients with chronic renal failure and a dialysis catheter. AB - This paper describes the incisional hernia repair technique carried out on 50 patients with chronic renal insufficiency requiring continuous ambulatory peritoneal dialysis. The technique involved fixing a polypropylene mesh prosthesis to the deep face of lateral incisions into the aponeurosis of the rectus abdominis muscle. Under general or epidural anaesthesia the old scar was resected and the hernial sac dissected, care being taken not to open the peritoneum. At 5 cm from either side of the hernia margin, the aponeurosis was incised longitudinally and the adjacent muscle separated. The mesh was sutured to the deep face of the aponeurosis, covering the hernial defect. A Tenckhoff catheter was left in the abdominal cavity. Peritoneal dialysis was given to all 50 patients in the immediate postoperative period, with no leaking of the dialysis fluid. One year later there has been no recurrence. Prosthetic mesh fixed to the deep face of the abdominal aponeurosis at incisions lateral to the hernia margins, without opening the peritoneum, is an effective treatment of incisional hernia repair for patients on peritoneal dialysis and allows early institution of dialysis. PMID- 11387730 TI - Comparison of severe life stress in depressed mothers and non-mothers: do children matter? AB - Given the high rates of maladjustment among children of depressed mothers, parenting is likely to cause significant life stress in this population, potentially worsening the course of mothers' depression. The present study is a comparison of severe life stress in 38 mothers and 62 non-mothers receiving treatment for recurrent major depression. Life stress was assessed using the Life Events and Difficulties Schedule [Brown and Harris, 1978a]. We hypothesized that mothers would evidence a greater number of severe life events and marked difficulties both in the year prior to the onset of their depressive index episode and in the time period following the onset of their current depressive episode. Prior to depression onset, mothers reported a significantly greater number of entrapping difficulties, but not marked difficulties, severe events, entrapping events, or humiliating events. However, following the onset of depression, mothers experienced a significantly greater number of severe events, entrapping events, marked difficulties, and entrapping difficulties, but not humiliating events. Mothers' elevated levels of stress were attributable to child related stress, predominantly related to children's psychological and behavioral problems. Our findings suggest that comprehensive treatment for mothers with major depression needs to address their parenting style and any psychological problems experienced by their children. PMID- 11387731 TI - Open trial of nefazodone among Hispanics with major depression: efficacy, tolerability, and adherence issues. AB - The efficacy and tolerability of nefazodone in the treatment of major depression among Spanish-monolingual Hispanics was examined and compared to historical controls among English-speaking, predominantly non-Hispanic subjects. Fifty monolingual Hispanic outpatients with major depression and a HAM-D17 score > or = 18 were treated with nefazodone in a flexible-dose 8-week open-label protocol. Sixty-three percent of the intent-to-treat (ITT) sample with > or = 1 efficacy visit were considered responders according to CGI-I criteria, falling within the range of response rates (58-69%) reported in six prior nefazodone trials with non Hispanic subjects. Significant improvement was found for the ITT and completer samples in HAM-D17, HAM-D28, and SCL-90 scores and in two measures of psychosocial functioning. Endpoint mean dose in the ITT sample was 379 mg/day (SD = 170), also within the range of previous trials (321-472 mg/day). Adverse effects were not elevated, with only dry mouth (8%) reported by > 6% of subjects. However, 42% of the sample dropped out of treatment before study termination, usually because of side effects or due to family or work difficulties, a higher rate than previously reported for nefazodone (21-33%). This open trial finds nefazodone to be an efficacious treatment for major depression among monolingual Hispanics, with comparable efficacy to previous controlled trials among non Hispanic subjects. Double-blind studies are required to confirm this comparable efficacy. Mean endpoint doses and adverse effect rates similar to previous trials do not support the need for reduced doses of nefazodone among Hispanics. However, an elevated rate of treatment discontinuation threatens treatment efficacy among this population. Causes for this elevated rate require explanation, given the apparently unremarkable pattern of adverse effect reports. PMID- 11387732 TI - Nonresponse to first-line pharmacotherapy may predict relapse and recurrence of remitted geriatric depression. AB - The authors examined whether nonresponse to first-line pharmacotherapy was associated with an increased probability of relapse or recurrence following remission of an episode of geriatric depression. The study group consisted of 74 elderly patients whose index episode of nonpsychotic unipolar major depression had responded to antidepressant pharmacotherapy. In 6 of these patients, the depressive episode had not responded to first-line pharmacotherapy (8 weeks of nortriptyline, including 2 weeks of adjunctive lithium) but it had responded to second-line treatment (phenelzine with or without adjunctive lithium). The 74 patients were maintained on acute doses of the medications that had led to response and were followed for 2 years or until relapse or recurrence, whichever occurred first. The cumulative probability of relapse or recurrence was 67% for patients who responded to second-line treatment compared with 18% for patients who responded to first-line treatment (P = 0.0003). As expected, mean time to response was significantly longer for patients who responded to second-line treatment but this factor did not account for the difference in outcome between the two groups. These findings suggest that pharmacotherapy resistance may constitute a risk factor for relapse or recurrence of remitted geriatric depression, even when patients are maintained on the medication that they eventually respond to. PMID- 11387733 TI - Clinician-administered PTSD scale: a review of the first ten years of research. AB - The Clinician-Administered PTSD Scale (CAPS) is a structured interview for assessing posttraumatic stress disorder (PTSD) diagnostic status and symptom severity. In the 10 years since it was developed, the CAPS has become a standard criterion measure in the field of traumatic stress and has now been used in more than 200 studies. In this paper, we first trace the history of the CAPS and provide an update on recent developments. Then we review the empirical literature, summarizing and evaluating the findings regarding the psychometric properties of the CAPS. The research evidence indicates that the CAPS has excellent reliability, yielding consistent scores across items, raters, and testing occasions. There is also strong evidence of validity: The CAPS has excellent convergent and discriminant validity, diagnostic utility, and sensitivity to clinical change. Finally, we address several concerns about the CAPS and offer recommendations for optimizing the CAPS for various clinical research applications. PMID- 11387734 TI - Community health. Clean hands rule. PMID- 11387735 TI - Hospital construction. Building booming. PMID- 11387736 TI - Outcomes reports. The grade debate. PMID- 11387737 TI - Medical training. Virtual ER. PMID- 11387738 TI - Managed care. Hassle healers. PMID- 11387739 TI - 1Q[3a]. Should patients be able to sue HMOs for malpractice? PMID- 11387740 TI - Help for rape victims. Dignity closet. PMID- 11387741 TI - Ambulance calls. Dispatch dispute. PMID- 11387742 TI - Outpatient surgery. Venturing out. PMID- 11387743 TI - BBA fallout. Suppliers feel the pinch. PMID- 11387744 TI - Voicing her opinion--with radio host Diane Rehm. PMID- 11387745 TI - A simple swap. You want investors' money; investors want better financial reporting. PMID- 11387746 TI - The assault on your financials. AB - Hospital executives are confronting what may be unprecedented pressure on their bottom lines as forces converge from all directions: Medicare cuts brought on by the Balanced Budget Amendment, slow pays and denial of claims by managed care plans, the staggering cost of new technology, and big increases in pharmaceutical costs. PMID- 11387747 TI - 1999 health care IPOs. PMID- 11387748 TI - Wish list 2000. Interview by Terese Hudson Thrall. PMID- 11387749 TI - Docs.com. PMID- 11387750 TI - Bioterrorism. PMID- 11387751 TI - Medication errors. PMID- 11387752 TI - [Interaction of Mycoplasma with immune system of animals and humans]. AB - Mycoplasmal infections of animals and human belong to latent or chronic ones, being commonly accompanied with immunodeficiency symptoms. The following topics are discussed in this review: 1) a direct influence of mycoplasmas on the immune system cells with an interference and compression of the functional activity of these; 2) influence of mycoplasmas on the immune system via cytokine induction; 3) mitogenes and mitogenic action of mycoplasmas; 4) the role of mycoplasmas in the development of autoimmune processes. Basic host immune response to mycoplasma is presented in diagrams. PMID- 11387753 TI - [Effect of cumulus and granulosa cells on meiosis resumption in murine oocytes in vitro]. AB - Effects of different follicular cell types on resumption of meiosis were studied. Cumulus enclosed oocytes (CEO), denuded oocytes (DO), cumulus cells (CCs) and mural granulosa cells (GCs) were used. Oocytes were obtained from mature gonadotrophin-stimulated and unstimulated mice. The resumption of meiosis was assessed by the germinal vesicle breakdown (GVBD) at the end of cultivation. It has been shown that GCs produced a meiosis activating substance due to gonadotrophin stimulation; for meiosis resumption connections between CCs and the oocyte were not necessary, but the very production of the meiosis activating substance, was, however, dependent on the initial connection between CCs and the oocyte. The presence of oocyte was necessary for stimulating CCs to produce a diffusible heat stable meiosis activating substance; gonadotrophins induced CCs to produce a diffusible thermostable meiosis activating substance. This substance induced, in a paracrine fashion, resumption of meiosis directly. It is proposed that the heat stable meiosis activating component of the used media from gonadotrophins-stimulated CEO may belong to a kind of meiosis activating sterols, previously isolated from human follicular fluid and from adult bull testes. PMID- 11387754 TI - [Relationship between the frequency of cultured oocytes fertilization and morphology of human sperm]. AB - The study has evaluated 304 cases under of vitro fertilization. Positive correlations were shown of semen parameters and fertilization rate. The individual spectrum of different sperm morphological types was shown to be constant. The discriminant analysis was used for comparison of two groups of patients--with low (less than 33%) and high (more than 67%) fertilization rates. The percentage of morphologically normal spermatozoa stronger correlated with fertilization rate. Although there was a fairly good correlation (in the order of their significance) with a progressive motility, grade of motility, percentage of slightly amorphous cells, sperm concentration, percentages of the neck and midpiece. The cut-off values of our discriminant test may conclusively predict the low (DF < 1.5) and the high (DF < 3.5) fertilization rates. PMID- 11387755 TI - [Ultrastructural localization of calcium pyroantimonate in Mauthner neurons of goldfish fry adapted to natural stimulation]. AB - The localization of Ca2+ in control and adapted goldfish fry Mauthner cells (M cells) revealed by sedimentation with potassium pyroantimonate technique was investigated. It has been shown the following. 1. In the control M-cells electron dense precipitates are present in the extracellular space, commonly within the active zone clefts of chemical synapses, throughout the whole apposition of the mixed synapses and in the synaptoplasm of both type afferent boutons. No precipitates were seen in the cytoplasm of M-cells. 2. After long term natural (vestibular) stimulation (LTNS), resulting in a strong functional suppression of M-cells, precipitates disappeared entirely from active zones but remained numerous in the cytoplasm of M-cells. The distribution of precipitates within the cytoplasm was non-uniform, the highest density was observed on the surfaces of intracellular organelles and elements of the cytoskeleton. 3. In fatigued M-cells after LTNS and after a subsequent one day rest the distribution of precipitates was less intensive, while in the whole it resembled that of fatigued M-cells. 4. In adapted M-cells the distribution of precipitates was similar to that observed in control. M-cells after LTNS, but the amount and size of the precipitated grains were noticeably increased. 5. The most numerous precipitates were seen in adapted M-cells after LTNS. They were localized throughout the postsynaptic cytoplasm and in a lesser order in the presynaptic cytoplasm. 6. After one day rehabilitation the intensitivity of cytochemical reaction of Ca2+ ion precipitation restored to the initial stage characteristic of adapted M-cells before LTVS. The results obtained suggest that the total concentration of Ca2+ ions in adapted M-cells and the dynamics of their exchanges between cytosole and intracellular depots, such as the smooth endoplasmic reticulum, may increase to keep a normal or even increased functional activity of M-cells, both before and after the LTNS. PMID- 11387756 TI - [Single-strand DNA breaks in brain cells of different rat strains under normal condition and during exposure to stress]. AB - This study was aimed to characterize pattern of occurrence of spontaneous single strand breaks in situ in glial and neuronal nuclei of the cortex, middle brain and hyppocapmi (CA3 field) of rats selected for a threshold of nervous system excitability, and to study the influence of stress of various modality on such breaks. The results obtained evidence that: 1) intact animals possess a subpopulation of glial and neuronal cells revealed following gap filling in situ in opposite to other types of terminally differentiated non-proliferating cells; 2) the size of such a subpopulation differs depending on the lines of examined rats, parts of brain, and the type of stress. PMID- 11387757 TI - [DNA degradation and repair in human laryngeal carcinoma HEp-2 cells after combined exposure to vitamin B12b and ascorbic acid]. AB - The formation and accumulation of DNA fragments containing no more than 23,000 pairs of bases were observed under exposure of human larynx epidermoid carcinoma cells (Hep-2) to "chemical nuclease", oxycobalamin (vitamin B12b) and ascorbic acid (vitamin C). The obtained DNA damages were repaired more slowly than those induced by gamma-irradiation in the dose adequate to the level of DNA damages. DNA reparation was not revealed after washing the cells from vitamin B12b and ascorbic acid, and in the course of cell incubation with ascorbic acid. Vitamin B12b and ascorbic acid separately did not induce degradation of DNA. DNA damages induced by "chemical nuclease" action precede the cell death observed later. PMID- 11387758 TI - [Expression of erythropoietin receptor mRNA in mouse brain hemispheres]. AB - Now there is a growing evidence that erythropoietin receptors (Epo-R) are present also in some nonhematopoietic tissues such as endothelial cells and fetal cells of neural origin, although the physiological role of Epo-R at these sites is unclear. There are some speculations that Epo-R may be expressed on cells only in the developing CNS. The objective of this study was to determine whether Epo-R mRNA may be expressed in the brain hemispheres of Balb/c mice of different age groups: 1) newborn mice, 2) young 2 months old mice, 3) old 1.8 year old mice. We also studied the in vivo effect of recombinant erythropoietin on the expression of Epo-R mRNA in the brain hemispheres of (CBA x C57BL)F1 mice by RT-PCR. We have detected the existence of Epo-R mRNA expression in brain hemispheres of all the groups, but in old mice this expression was significantly higher. We have discovered a decrease in Epo-R mRNA expression in brain hemispheres of (CBA x C57BL)F1 mice 24 h after in vivo administration of recombinant erythropoietin. The Epo-R mRNA expression in the left brain hemispheres of (CBA x C57BL)F1 was considerably higher than in the right one. PMID- 11387759 TI - [Cytological aspects of similarity and difference of Myxozoa and Cnidaria]. AB - A comparative cytomorphological analysis of Myxozoa and parasitic Cnidaria Polypodium hydriforme has been carried out in view of the Weill (1938) hypothesis, which regards Myxozoa as a reduced Cnidaria. The question on the relation of Myxozoa and Cnidaria was arising several times with the application of some new methods during the Myxozoa studies. At present the idea on their phylogenetic relationships has appeared again in connection with an absolutely new understanding of the myxozoan life cycle (Wolf, Markiw, 1984), as well as with the application of molecular-biological methods for their phylogenetic studies. The latter, however, provided some diverse results. So far no comparative cytomorphological analysis of Myxozoa and Polypodium has been carried out. The present paper is to fill the gap on the basis of accumulated facts. According to Weill (1938), the features of similarity of parasitic Cnidaria and Myxozoa are the following: 1) the presence in both of extrusomes (nematocysts and polar capsules) whose structure and development are surprizingly similar; 2) the nuclear dimorphism and somato-generative segregation; 3) the presence of a somatic nutritional cell, surrounding the multiplying generative cells; at present it is known that polyploidy of somatic nuclei and the absence of parasitophorous vacuole are characteristic of trophamnion of Polypodium and trophozoite of Myxozoa; 4) the presence of radial symmetry in both groups; 5) the construction of a diblastic organism made of a cluster of endodermal cells and a few ectodermal cells; 6) the similarity of their cell contacts (Grasse, 1970). At present it is possible to add to Weill's (1938) list of features common for parasitic Cnidaria and Myxozoa the number of important similarities between Polypodium and Myxozoa, some of which being not characteristic of Cnidaria: 1) the "cell in cell" organization of all Polypodium parasitic stages and all Myxozoa life cycle stages; 2) the presence of gametophore supplied with extrusomes; 3) both organisms have haplophase in their life cycles preceded by two-step meiosis; 4) there are mitochondria with tubular cristae in both organisms; 5) the absence of spermatozoa and eggs in both organisms; 6) the similarity of Polypodium cnidocile structure and the cone-like formation situated at the anterior end of polar capsule of actinospore (Lom. Dykova, 1997); 7) the participation of MTOC in the formation of extrusomes in both animals. In spite of the obvious similarity between Myxozoa and parasitic Cnidaria (including Polypodium) it is, however, necessary to take into account differences between them, the main being as follows: the absence in Myxozoa of flagellated stages, centrioles, tissues and organs, true blastophylla, planula-like larvae, gastrulation; the presence of low cell integrations in Myxozoa; Cnidaria and Myxozoa have different types of mitosis, their life cycles and the discharge mechanism of their stinging apparatus being also different. We consider as quite valid a suggestion by Siddall et al. (1995) that parasitic Cnidaria could present an early separated branch of the cnidarian evolution. Further studies of Myxozoa life cycle may show their more definite relation to parasitic Cnidaria. The problem has not yet been solved completely since the available molecular biological data are rather contradictory and moreover there is no distinct idea as to the Eumetazoa ancestor so far. A further thorough investigation is badly needed in the feelds of developmental cycle, cytomorphology and molecular biology of the variety of narcomedusae and representatives of Myxozoa. This may help to find some transitional forms and stages of the animals and to understand whether we deal with a regressive evolution of parasitic Cnidaria or with a parallel evolution of taxa originated from the common ancestor. PMID- 11387760 TI - [Current role of antidopaminergic drugs in pediatrics]. AB - The treatment of gastrointestinal disorders in pediatrics is based on the use of prokinetic agents; amongst these prokinetic compounds, cholinomimetic drugs and dopamine antagonists (metoclopramide, domperidone) are principally available. Metoclopramide is an antidopaminergic benzamide with mainly antiemetic effects, due to the interaction with dopamine receptors in the central trigger zone. Another effect is to enhance the transit of material through the gastrointestinal tract. Disadvantages in the use of metoclopramide are neurological effects (asthenia, sleepiness), extrapyramidal dyskinetic reactions and/or neuroendocrine side effects (galactorrhea). Domperidone is a pure dopaminantagonist that accelerates gastric emptying. It is useful in the treatment of acute nausea and vomiting caused by different agents. There are adverse effects only if it is used in parenteral way. PMID- 11387761 TI - Megaureter: classification, pathophysiology, and management. AB - The term megaureter does not define a specific pathological condition, because it can be due to different underlying abnormalities. The most used classification includes three groups: refluxing megaureter, associated with vesicoureteral reflux (VUR); obstructive megaureter, associated with urine flow impairment at the vesicoureteral junction; non-refluxing non-obstructive megaureter, if neither obstruction nor reflux can be identified. Each group can be divided into two subgroups: primary megaureter; secondary megaureter. With the advent of antenatal ultrasound an increased number of cases are identified prior to the onset of symptoms. The common used investigation are: urinary tract ultrasound, voiding cystourethrography, urography, serial diuretic renography and pressure-perfusion studies (Whitaker test). The advent of prenatal and neonatal echography has modified the natural history of megaureter. Nowadays non operative management is preferred. Operative intervention is indicated only in these cases: significant impairment to urine flow; worsening renal function during the observation time; recurrent UTI in spite of adequate antibiotic prophylaxis. PMID- 11387762 TI - [Etiology and precipitating factors of cyclic vomiting]. AB - The aim of the study was to determine the nature, severity, precipitants and associated features of attacks and the incidence of potential aetiological factors of cyclic vomiting syndrome (CVS). Ten patients (6 boys and 4 girls aged 2-12 years) with CVS, defined as recurrent episodes of vomiting with symptom-free intervals occurring two or more times per year, with episodes having a similar pattern and for which no other cause could be found, ten patients (7 boys and 3 girls aged 7-14 years) with migraine, defined as recurrent headache with symptom free intervals and at least three of the following symptoms or associated findings: abdominal pain, nausea or vomiting, throbbing headache, unilateral location, associated aura (visual, sensory, motor), relief after sleeping and ten controls (6 boys and 4 girls aged 4-13 years) were studied. The mean age at onset of symptoms in patients with CVS was 3.9 years and the mean number of attacks per year was 4, the average duration of episodes was 20.5 hours and the mean number of school missed days were 6 per year. The mean age at onset of symptoms in patients with migraine was 7.6 years and the mean number of attacks per year was 9.9, the average duration of episodes was 8 hours and the mean number of school missed days were 6.4 per year. A family history of migraine was significantly higher in both groups of patients, compared with controls (p = 0.009). A personal history of headache and/or migraine and associated sign or symptoms like recurrent abdominal pains, limb pain and kinetosis was significantly higher in both patients compared with controls (p < 0.001). CVS is a chronic, disabling condition and is a migraine variant, with attacks usually precipitated by stress and intercurrent infections. PMID- 11387763 TI - Management of idiopathic thrombocytopenic purpura in children: a single institution experience. AB - Idiopathic thrombocytopenic purpura (ITP) is a relatively common hematologic disease in childhood. ITP is often self-limiting and is characterized by a good clinical outcome. About 10-20% of cases can have a chronic course. In our retrospective analysis we have evaluated 45 patients affected by ITP from January '92 to December '97. Thirty-seven patients (82%) met the criteria of acute ITP and 8 (18%) had chronic ITP. Patients were stratified into 3 categories based up on the type of treatment received: no treatment, steroids, steroids and IVIG. In our series children treated with oral prednisone showed a slightly faster recovery in the first days from treatment. We suggest the use of steroids in children with low platelet count and signs and symptoms of bleeding. PMID- 11387764 TI - [Predictive factors of etiology (bacterial or viral) in acute gastroenteritis in childhood]. AB - The aim of the study was to identify immediate clinical and/or laboratory findings able to differentiate bacterial from viral etiology of acute gastroenteritis in pediatric patients. We studied 52 children, aged between 5 months and 12 years, consecutively admitted to hospital with acute diarrhoea lasting less than 5 days. All the patients were divided into 4 groups according to etiologic agent, subsequently demonstrated by culture: salmonellae (group A), rotavirus (group B); combined salmonellae and rotavirus (group C) and no pathogen (group D). The contemporary presence of fever > 39 degrees C, number of daily liquid stools > 6, presence of bloody diarrhoea, positivity of C-reactive protein and hyponatremia (< 135 mEq/l) allowed to recognize the etiology (viral or bacterial) before results of culture (sensitivity was 71% and specificity was 97%). In particular, hyponatremia resulted significantly lower in group A and C than in group B and D. We concluded that hyponatremia can be considered a marker for acute gastroenteritis caused by salmonellae. PMID- 11387765 TI - [Long-term follow-up of chronic idiopathic thrombocytopenic purpura in children]. AB - Idiopathic thrombocytopenic purpura (ITP) is a common acquired bleeding disorder in infancy and childhood. Most children rapidly improve, exhibiting a rise in platelet count to hemostatically normal levels within weeks to several months. Traditionally, chronic ITP is defined as persistence of thrombocytopenia (platelet count < 150 x 10(9)/L) for greater than 6 months. The Authors retrospectively evaluated 16 patients with chronic ITP, identified during a 12 year period of time in their Department of Pediatrics. The most important clinical and hematological parameters of patients were analyzed, including age at diagnosis, type and response to the initial treatment, number of multiple treatments, and duration of follow-up. At the last evaluation (december 1999) one patient was lost to the follow-up; one died of overwhelming postsplenectomy sepsis; four still require intermittent or chronic infusions of intravenous gamma globulin; seven are in stable partial remission (PLT > 50 x 10(9)/L < 150 x 10(9)/L) and do not require any treatment; three are in complete remission (PLT > 150 x 10(9)/L). Finally, the Authors discuss of the natural history and management of this rare disease. Presently there are insufficient trial data to support evidence-based treatment guidelines in childhood chronic ITP and therefore it is reasonable to encourage future multicentre collaboration. PMID- 11387766 TI - [Kearns-Sayre syndrome with GH deficiency]. AB - A case of Kearns-Sayre syndrome with failure to thrive, cytochrome oxidase deficiency in muscle fibers, mtDNA deletion and deficient GH responses to stimulation tests is reported. The patient, when 9 years old, was treated with GH without significant improvement in growth velocity. The opportunity of search for GH deficiency in children with Kearns-Sayre syndrome and failure to thrive is stressed. PMID- 11387767 TI - [Intractable diarrhea and neuroblastoma: report of a clinical case]. AB - One of the typical presentations of neuroblastoma is intractable diarrhea or wdha (watery diarrhea, hypokalemia, achloridria). The case admitted to our Pediatric Surgery Department presented watery diarrhea due to VIP hyperincretion caused by a stage 1 neuroblastoma, whose ablation allowed a complete resolution of the clinical conditions. This case report can be useful in the discussion of the differential diagnosis of the most common clinical pictures. PMID- 11387768 TI - [Pyloric hypertrophic stenosis in the premature child. A clinical case]. AB - Hypertrophic pyloric stenosis (HPS) is rare in premature infants. We report a case of HPS in an extremely low birth weight neonate (28 weeks of gestation, 622 gr), discovered on the 10th week of life. Although the diagnosis and treatment of pyloric stenosis has ben well established for many years, the presentation in preterm babies is atypical and the diagnosis often delayed. A brief review of literature is included. PMID- 11387769 TI - [A severe case of Plasmodium falciparum malaria]. AB - Malaria infections has been an important public health problem in Europe and Western countries in the last years. The international travels and moreover immigration from Africa and Asia countries has been rapidly increasing especially in Italy. The transmission and diffusion of malaria has increased, especially over last decade, due to vector resistance to insecticides and chemoresistance of these parasites to most antimalarial drugs. The authors presents a severe case of malaria infection that was caused by Plasmodium Falciparum in a one year-old female children, born in Italy, infected during a return in the parents' country of origin, Nigeria. She was admitted because of febrile gastroenteritis for five days. Since the child was in a good state of health, normal the medical examination, blood tests and radiography of the chest. The next day, persisting the fever, the child was somnolent, pale, and was present a haemoglobin concentration of 5.1 g/dl and a thrombocytopenia, a complicated falciparum malaria was diagnosed (8% parasitaemia). Treatment was started immediately with a single oral dose of Mefloquine (25 mg/Kg). Red blood cells were transfused once. The parasitaemia dropped to 4% and 0.8% in less than 48 hours. Weekly controls for the following four weeks remained negative. PMID- 11387770 TI - [PCA in the control of acute and chronic pain in children]. AB - BACKGROUND: This study was undertaken to verify the efficacy and safety of patient controlled analgesia (PCA) in preschool and older than 6 years children with acute postoperative and chronic oncologic pain. METHODS: Sixty-two children, aged 4-17 yr, with postoperative (n = 37) or chronic (n = 25) pain received PCA with morphine at the following dosages: bolus = 10-20 gamma kg-1; starter dose = 10-30 gamma kg-1; background infusion = 10-50 gamma kg-1 h-1. Lockout was 5 to 15 minutes. In some children with oncologic pathology adjuvant drugs were also used. RESULTS: Analgesia was considered satisfactory in all children, while a mild sedation was observed only in two children. PCA was also successfully used in preschool children (11%), aged 4 to 6 yr. Side effects were observed in 8 patients, but only in one of them a transient respiratory depression was recorded. The mean PCA duration was 5.3 +/- 5.5 days (min.2; max 24). CONCLUSIONS: These results indicate that PCA is an effective and safe method to control acute postoperative pain as well as to manage chronic oncologic pain in children. PMID- 11387771 TI - [Anterior cervical corpectomy in the treatment of multilevel degenerative stenoses with spondylotic myelopathy. Personal experience with therapy and a literature review]. AB - The authors use as a basis experience with a group of 389 patients operated in 1989-1997 on account of degenerative diseases of the cervical spine with neurological manifestations. The results are compared with experience assembled in 1998-1999. In the new group (188 patients with the same diagnosis) the same indication criteria were used but in case of myelopathy associated with multisegmetal cervical stenosis not only multilevel discectomy was performed (as in the previous group) but in addition also 1-3 segmental corpectomy (somatectomy). The results were evaluated separately for both methods used. The authors emphasize the necessity of a radical approach during decompression of neurological structures incl. removal of uncovertebral osteophytes which must be combined with suitable stabilization of the fusions or possibly be supported by instrumentation. In the discussion the authors illustrate in the form of a review the development of anterior cervical corpectomy as one of the methods of an anterior approach to the cervical spine used at first in traumatic and oncological conditions, later extended to operations on account of degenerative, dysplastic conditions and other diseases. The authors wish to help to define criteria for application of corpectomy when treating stenoses of the spinal canal in the cervical region. The indication will be defined in a perspectively followed up group where a detailed clinical and electrophysiological algorithm for examination was submitted as well as postoperative follow-up and processing of the resulting data. PMID- 11387772 TI - [Surgical treatment of thyroid gland carcinoma--a 5-year retrospective study]. AB - The authors analyze a group of 916 patients who had thyroid operations in the course of five years at a Surgical Department in Bratislava. 256 patients suffered from malignant disease. The authors analyze the incidence of different histological types of carcinoma, types of surgical operations, complications and patient survival. They discuss the problem of thyroid microcarcinoma, the relationship of Hashimoto's thyroiditis and carcinoma. The authors emphasize the importance of exact implementation of the first operation of the thyroid gland, in their opinion leaving residues of the gland during Berry ligature is inadequate. Re-operations should be implemented by departments with a high professional standard of surgeons and standardized pre- and postoperative patient care, in collaboration with an endocrinologist, pathologist and a department of nuclear medicine. PMID- 11387773 TI - [Prophylactic use of counterpulsation in patients with severe myocardial dysfunction]. AB - During the period between 2/2000 and 11/2000 21 patients with severe ventricular dysfunction were investigated (EF less than 30%) where in a planned manner intraaortic balloon counterpulsation was introduced before cardiac surgery. The low mortality (5%), zero incidence of complications caused by the method and the improvement of the contractility of the heart muscle, evaluated from a rise of the ejection fraction (EF) make so-called prophylactic use of contrapulsation possible as a method of first choice in severe myocardial dysfunction. PMID- 11387774 TI - [Esophageal atresia: delayed surgical treatment of the long segment]. AB - Atresia of the long segment of the oesophagus in children and its surgical treatment is frequently very complicated and is associated with a high rate of complications. The authors present a patient with a long segment of oesophageal atresia type Vogt IIIB. The distance between the oral and aboral stump was 4 cm. During the first session the authors made a thoracotomy, separated the aboral stump from the trachea and closed the opening in the trachea as well as the distal stump. Then they approximated both stumps without attempting an anastomosis and fixed them by stitches to the spine. They closed the chest and performed a gastrostomy. After eight weeks they repeated the thoracotomy and made an oesophago-oesophago end-to-end anastomosis. The patient was subjected after operation three times to balloon dilatation on account of a stricture at the site of anastomosis. He is now 18 months after operation and has no complaints and no stricture. PMID- 11387775 TI - [Bacterial flora in obstructed bile ducts]. AB - The authors present the results of bile cultivation from obturated bile ducts. The bile was obtained during ERCP. The results agree in principle with data reported in the literature. The authors emphasize the regional variability of bacterial strains and their sensitivity. It is thus an ever urgent problem which deserves attention, in particular preventive administration of antibiotics. At the authors department as preventive antibiotic cefoperazone proved useful. PMID- 11387776 TI - [Incidence of inguinal hernias]. AB - Groin hernia operation is the third most frequent operation in the Czech republic. Early recurrence of the hernia after prime operation is a medical failure. The recurrences can arise after all types of operation. Incidence of recurrences, time factors of its genesis and treatment are discussed there. PMID- 11387777 TI - [Basic principles of surgical treatment of recurrent inguinal hernias]. AB - The incidence of the recurrences after groin hernia operation can be surprisingly high. Both basic principles of the surgical treatment (tension on, tension free) of this disease are known more than 30 years. It seems, that the latest endoscopic-laparoscopic proceedings have smaller recurrences. However long-term results are absent for more than 10 years from the prime surgery. In the report the basic principles of prime groin hernia surgery are mentioned, both classical transinguinal operation and endoscopic surgery. In the end the fundamental scheme is introduced--how to proceed in surgical treatment of groin hernia recurrences. PMID- 11387778 TI - [Iatrogenic injuries of the colon during colonoscopy]. AB - Endoscopic examinations of hollow organs, similarly as other invasive operations, are associated with a certain percentage of sometimes very serious complications. Iatrogenic perforations of the gut, in particular during the procedure belong to the most dramatic ones. During the period of 1990-2000 the authors had the opportunity to deal in collaboration with the medical department with five cases. PMID- 11387779 TI - [The first kidney block transplantation in the Czech Republic--results]. AB - A case report of the fate of the first cadaveric pediatric kidney block transplantation into adult recipient in the Czech republic. The operation was done in Regional transplant centre in Faculty hospital Ostrava in February 1994. The whole postoperative period has been recapitulated since the operation to the end 2000. Pediatric kidney graft is functional 7 years after transplantation. PMID- 11387780 TI - [Results of surgery in children with congenital megacolon]. AB - In 1974 to 1999 at the Department of Paediatric Surgery in Bratislava 142 patients aged 3 days to 14 years were treated on account of Hirschsprung's disease. Seventy-four patients suffered from the classical type of Hirschsprung's disease affecting the recto-sigmoid segment of the large bowel, in 43 the long segment of the large bowel was affected. In nineteen patients an ultrashort segment was afflicted and six patients suffered from total aganglionosis of the large bowel. In 131 patients the diagnosis was established by irrigographic examination and during operation it was confirmed by histological examination of the whole month of the large intestine. In 11 patients the diagnosis was established by peroperative collection of the whole month of the large bowel. In 43 patients the diagnosis was made during the neonatal period. All 142 patients were operated. Nineteen patients with an ultrashort segment were subjected to dorsal myectomy according to Lynn and in 121 patients a retrorectal pull-through according to Duhamel in Ikeda-Soper's modification was performed. In 99 patients before the final operation a colostomy was made, orally from an aganglionic portion of the large bowel. Twenty-two patients were treated in one session by means of a direct stapler without protective colostomy. Two patients died several days after establishment of the colostomy from anaerobic septic complications. After the final operation no death was recorded. Duhamel's operation in Ikeda Sopor's modification is in the authors' opinion very effective in treatment of congenital megacolon, being associated with a minimum of complications and is the prerequisite for achieving excellent functional results. PMID- 11387781 TI - [Perioperative hypothermia and its sequelae]. AB - Perioperative hypothermia is associated with the development of haemocoagulation, cardiovascular and metabolic disorders leading to an increased morbidity and mortality. The objective of the investigation was to assess the extent of hypothermia and its clinical and laboratory consequences. A group of 30 patients subjected to elective radical laparotomy on account of colorectal carcinoma was divided into to equivalent groups. To the first group heated infusions were administered, to the second group not heated ones. In all patients the central and peripheral temperature, rate of postoperative normalization of the temperature, postoperative thermal comfort, consumption of analgesics and biochemical and haematological parameters were monitored. RESULTS: In patients with non-heated infusions more marked and longer perioperative hypothermia was recorded with a significant alteration of the number of leucocytes and thrombocytes. In the other investigated indicators there was no significant difference between the two groups. Hypothermia did not cause serious complications in any of the patients. CONCLUSION: Although no serious clinical complications induced by hypothermia were recorded, the authors recommend an active approach and provisions for the perioperative maintenance of body temperature as a standard of contemporary perioperative care. PMID- 11387782 TI - [Effect of carbon dioxide pneumoperitoneum on selected parameters of the acid base equilibrium in laparoscopic cholecystectomy]. AB - The authors focused attention on a hitherto less well known area of the effect of capnoperitoneum during laparoscopic cholecystectomy on homeostasis of the internal environment, i.e. values of the acid-base balance and its changes during surgery and the early postoperative period. From a randomized perspective study ensued that intraabdominal pressure and the reduction of the intraabdominal CO2 pressure from 15 mmHg to 10 mmHg did not have a significant effect on parameters of the acid-base balance in the investigated group of patients as compared with a control group. From the results it is apparent that possible danger of acidosis as a result of CO2 absorption from the abdominal cavity, deterioration of the splanchnic blood supply or a negative impact of the intraabdominal overpressure on the cardiovascular apparatus is sufficiently compensated by the respiratory system during artificial pulmonary ventilation. There is however a certain danger of acidosis during the immediate postoperative period after extubation when the patients are not yet able to compensate possible fluctuations of the acid-base balance and are according to the authors' measurements in a state of mixed or respiratory acidosis. But even in this respect no statistically significant difference was observed between the two groups. These assessments were made in relatively healthy patients, i.e. group I and II according to the ASA classification, therefore these results will apply even mor in groups of risk patients of higher ASA categories. In particular in these patients it is useful to adjust the insufflation pressure individually to lower values in order to reduce to a minimum possible negative impacts of the capnoperitoneum. PMID- 11387783 TI - [Surgical activity in emergency health services (analysis of activity at the Trauma Hospital in Brno in 1990)]. AB - In the Traumatological Hospital in Brno in 1999 4975 operations were made incl. 1064 (21%) during emergency services. Thus there are on average 3 (2.91) operations per emergency shift. 88 operations were made after midnight. The operations implemented during emergency shifts can be divided into three groups: urgent operations, operation on the common schedule which were delayed but partly also non-urgent operations which are performed because of economic pressure. The legal changes limiting overtime work make it important to seek ways how to preserve under the new conditions the scope of surgery while it is assumed that there will be a greater pressure to refer patients to the centre. The financial conditions of the Institute as regards funds are at variance with the efforts made. PMID- 11387784 TI - [A patient with spontaneous healing of traumatic spondylolisthesis of the cervical vertebrae]. AB - The authors submit a report on a patient with traumatic spondyloptosis of the sixth cervical vertebra which occurred in an adult man when he bumped his head against a standing lorry. The injury of the cervical spine was diagnosed only four weeks after the accident when the patient regained consciousness and the vertebrae were already fixed in a pathological position. Seven years after the injury the patient is satisfied with his health, works in his original occupation, is engaged in sports and leads an active life. PMID- 11387786 TI - Clinically significant differences among Canadian mental health acts. AB - OBJECTIVES: First, to highlight the major differences among mental health acts in different Canadian jurisdictions as they relate to involuntary admission criteria, treatment authorization, review and appeal procedures, and conditional leave and community treatment orders. Second, to analyze the impact of these differences on the care that individuals with mental illness receive. METHOD: We examined the mental health act provisions of all Canadian jurisdictions to determine how the clinical management of a typical case would differ among jurisdictions. We used a statement of principles for mental health legislation endorsed by the Canadian Psychiatric Association to guide the analysis. We confirmed interpretation of each act and its implementation through key informant contact in each province and territory. RESULTS: We found clinically significant differences among the provinces and territories on all major components of their mental health acts. CONCLUSION: Provisions that prevent patients receiving appropriate clinical care can be found in some Canadian mental health acts. Alternate provisions that support appropriate clinical care, that respect the human rights and personal dignity of patients, and that are consistent with the Canadian Charter of Rights and Freedoms can be found in the legislation of other jurisdictions. PMID- 11387787 TI - Lithium revisited. AB - OBJECTIVE: To review lithium's utility in the treatment of mood disorders. METHOD: We reviewed the safety, tolerability, teratogenicity, optimal dosing regimens, and mortality-lowering effects of lithium. Clinical relevance and scientific rigour determined which articles we selected for review. RESULTS: Lithium is the paradigmatic treatment for bipolar disorder (BD). In treating BD, optimal maintenance plasma levels may be approximately 0.75 to 0.85 mEq/L. Although nephrogenic diabetes insipidus is not uncommon, irreversible renal failure due to lithium appears to be a rare, idiosyncratic event. Lithium-induced cardiovascular teratology appears to be less common than previously thought. Optimal lithium dosing may be once daily, this agent appears to bestow a robust suicide-lowering effect, and emerging data hint at neurotrophic and neuroprotective effects. CONCLUSION: Lithium remains an effective and integral agent in the treatment of BD. Its ability to lower suicide rates in persons with BD warrants clinical attention. PMID- 11387788 TI - Lamotrigine: a review of clinical studies in bipolar disorders. AB - OBJECTIVE: This article reviews published studies on the use of lamotrigine in the treatment of bipolar disorder (BD). METHOD: We performed a Medline search to identify the literature data base available on double-blind, open-label studies and case series on the use of lamotrigine to treat BD. RESULTS: Three double blind studies, 3 open-label studies, and 2 case series have been conducted to date (n = 401 patients). Most patients were either nonresponders or partial responders to other mood stabilizers. Overall, 50% to 83% of the patients responded to lamotrigine; doses in the studies ranged from 50 to 400 mg daily. Switching to mania while on 200 mg of lamotrigine or more was extremely rare, and there were no reports of serious adverse effects during the study periods. CONCLUSION: Lamotrigine is proving to be an effective agent in the treatment of BD and may be useful for patients who have not responded to other mood stabilizers. PMID- 11387789 TI - Antipsychotic polypharmacy: a survey of discharge prescriptions from a tertiary care psychiatric institution. AB - OBJECTIVE: To perform a retrospective survey of discharge medications at a tertiary care psychiatric facility and to assess the incidence of antipsychotic polypharmacy. METHOD: This is a retrospective survey that used the Department of Pharmacy's computer database to obtain relevant discharge information on all nongeriatric patients with schizophrenia discharged from Riverview Hospital between November 1, 1996 and October 31, 1998. From a total of 492 eligible patients, 229 met the inclusion criteria and formed the database for the survey. RESULTS: The main finding of the survey was that 27.5% of our discharged patients diagnosed with schizophrenia left our facility on an antipsychotic polypharmacy regimen. Compared with patients discharged on a single antipsychotic, the pooled data revealed a significantly greater use of anticholinergic agents in those patients prescribed an antipsychotic polypharmacy regimen. Further, of the atypical agents, only risperidone showed a statistically significant reduction in dosage when coprescribed with a second antipsychotic. CONCLUSION: Although antipsychotic polypharmacy persists today, as it has over the past 30 years, evidence-based data to support this controversial treatment strategy is lacking. As a result clinicians are relying on their clinical experience, and perhaps intuition, to design antipsychotic polypharmacy treatment protocols. Efforts should be made to replace subjective clinical impression with a more rational approach to antipsychotic polypharmacy--one that is based on pharmacodynamic and pharmacokinetic understanding of drug action. PMID- 11387790 TI - Regional cerebral glucose metabolism in never-medicated patients with schizophrenia. AB - OBJECTIVE: The purpose of this study was to assess regional cerebral glucose metabolism in patients with schizophrenia who had never received antipsychotic medication and whose olfactory identification ability had been assessed. Two hypotheses were examined. First, the patients were compared with normal controls to determine whether differences in regional cerebral metabolism were apparent. Second, regional rates of metabolism were correlated with olfactory ability and the relation between them determined. METHODS: The patient (n = 26) and control (n = 32) subjects were scanned at rest using positron emission tomography (PET) after administration of 18F-fluorodeoxyglucose (FDG). In addition, the University of Pennsylvania Smell Identification Test was administered to each patient. RESULTS: Patients with schizophrenia had reduced rates of glucose metabolism in the right and left thalamus that reached significance if not corrected for multiple comparisons. However, if a Bonferroni correction was applied over the 27 regions of interest, the differences were not significant. Scores on the Smell Identification Test were negatively correlated with 8 regions of interest. When scores were analyzed using multiple regression, the left frontal cortex and the medial parietal cortex were significant predictors. CONCLUSIONS: The finding of reduced metabolism in the thalami is consistent with some of the previous literature, whereas the negative correlations between specific regions and olfactory function are not consistent with studies using activation paradigms. PMID- 11387791 TI - A family study of juvenile obsessive-compulsive disorder. AB - OBJECTIVES: To determine whether juvenile obsessive-compulsive disorder (OCD) is familial and whether the rate of Tourette syndrome (TS) and tic disorders is higher among relatives of patients with OCD than among relatives of controls subjects. METHOD: We assessed first-degree relatives of 35 juvenile OCD probands (aged 16 years or less) and 34 matched, psychiatrically unaffected control subjects, using the Diagnostic Interview for Children and Adolescents-Revised (DICA-R) (unpublished), a Questionnaire for tic disorders, the Children's Version of Leyton's Obsessional Inventory (CV-LOI), and the Children's Version of the Yale-Brown Obsessive Compulsive Scale (CY-BOCS). Similarly, we assessed adult relatives, using the Schedule for Clinical Assessment in Neuropsychiatry (SCAN), Leyton's Obsessional Inventory (LOI), the Yale-Brown Obsessive Compulsive Scale (Y-BOCS), and a Questionnaire for tic disorders. The diagnoses were determined by consensus, using DSM-III-R criteria. We calculated age-corrected morbid risk, using Weinberg's method. RESULTS: The morbid risk for OCD among the relatives of OCD probands was 4.96%, while none of the relatives of unaffected control subjects had OCD. We did not diagnose TS in any of the relatives of either OCD probands or control subjects. We diagnosed chronic motor tic disorders in only 1 of the relatives of OCD probands, while none of the relatives of control subjects had any tic disorder. CONCLUSION: Most juvenile cases of OCD are nonfamilial and unrelated to tic disorders, while only a few are familial. There is a need to re examine the issue of familiality in cases of OCD, as well as its relation to TS, using larger community samples to better understand the hypotheses of familial transmission and comorbidity with tic disorders. PMID- 11387792 TI - Clinical trials in psychiatry: do results apply to practice? AB - OBJECTIVE: The purpose of this communication is to alert psychiatrists to the difficulties of translating results of group difference obtained from large, randomized clinical trials to the treatment of individual patients. METHOD: Reported discrepancies between a) clinical trial participants and general psychiatric patients, b) clinical trial investigators and general clinicians, and c) study trial and usual clinic conditions were assessed. RESULTS: The results confirm that important differences exist in all 3 areas. CONCLUSIONS: Recommendations for researchers include more complete assessments of factors that account for individual difference, an appraisal of outcomes more important to patients than symptom scores, and the use of statistical methods that permit the evaluation of individual difference. Recommendations for clinicians include a careful differentiation of results obtained in different phases of clinical trials and a clear appreciation of the different purposes of those trials. Clinicians should also appreciate that short-term effectiveness is not the same as long-term outcome and that aggregate scores may not apply to individual patients. PMID- 11387793 TI - Olanzapine augmentation of serotonin uptake inhibitors in obsessive-compulsive disorder: an open study. AB - OBJECTIVE: This study evaluates the clinical response to olanzapine used in association with selective serotonin reuptake inhibitors (SSRIs) or clomipramine in treatment-resistant obsessive-compulsive disorder (OCD). METHODS: We describe the cases of 9 patients with serotonin uptake inhibitor-resistant OCD who were given an open-label adjunctive treatment of olanzapine for a minimum of 6 weeks. The response was assessed by using the Yale-Brown Obsessive Compulsive Scale (Y BOCS) and the Clinical Global Impression Scale (CGI). RESULTS: Six patients showed improvement of symptoms after the augmentation with olanzapine. One patient (treated with clomipramine) discontinued olanzapine due to side effects, and another 2 did not respond. CONCLUSION: Olanzapine augmentation of SSRIs in treating OCD showed a good (two-thirds) response rate, and it could therefore be considered as a treatment option when conventional therapies have failed. PMID- 11387794 TI - The effect of divalproex sodium on viral load: a retrospective review of HIV positive patients with manic syndromes. AB - OBJECTIVE: In vitro studies report that valproic acid causes an increase in HIV-1 replication. This retrospective study examines a sample of patients with HIV disease and behavioural disturbances, for which the treatment of choice is divalproex sodium (DVP), to determine whether DVP causes an increase in HIV-1 replication. METHOD: A chart and database review identified 15 patients with HIV disease presenting with either 1) mania or hypomania or 2) dementia with mania or with hypomania or behavioural disturbances. HIV-1 viral load was compared before and after mood stabilizer initiation. RESULTS: Eleven patients started therapy with DVP, and 4 patients declined treatment with a mood stabilizer. Nine of the 11 patients taking DVP were also receiving antiretroviral therapy. HIV-1 viral load did not increase in 6 of the 9 patients who had measurements between 1 week and 3.5 months after DVP initiation. No follow-up was available for the other 3 patients. Of the 2 patients receiving DVP but not antiretroviral medication, 1 had an increase of 0.17 log in HIV-1 viral load at 4 months. No follow-up record was available for the second patient. The 4 patients not taking DVP were all on antiretroviral therapy; viral loads in 2 of them remained nondetectable over 3 to 4 months, and 1 had an increase of 0.32 log in HIV-1 viral load at 3 months. No follow-up record was available for the fourth patient. CONCLUSIONS: These preliminary data suggest that, in the presence of effective antiretroviral therapy, HIV-1 viral load appears not to be adversely affected by the administration of DVP. The results for the patients receiving DVP in the absence of antiretroviral medication remain indeterminate. Further prospective study is required. PMID- 11387795 TI - Re: Neuroleptic malignant syndrome associated with quetiapine. PMID- 11387796 TI - Neurobiology of the use of selective serotonin reuptake inhibitors during pregnancy. PMID- 11387797 TI - Ondansetron rather than metoclopramide for bupropion-induced nausea. PMID- 11387798 TI - The use of intravenous valproate in psychiatry. PMID- 11387799 TI - Sudden death following neuroleptic administration due to hemoperitoneum resulting from physical restraint. PMID- 11387800 TI - Does the cellular phone help to communicate when face-to-face contacts are difficult? PMID- 11387801 TI - What factors contribute to senior psychiatry residents' interest in geriatric psychiatry? PMID- 11387802 TI - A need for clarity in funding spiritual interventions. PMID- 11387803 TI - Ophthalmic manifestations of HIV: an update. AB - A variety of ocular disease processes have been identified in HIV-antibody positive persons, especially in people with AIDS. The most common sight threatening disease in this population is cytomegalovirus (CMV) retinitis. Effective therapies are available to treat CMV retinitis. However, such therapies carry potentially challenging toxicities. Highly active antiretroviral therapy (HAART) has improved immune system functioning in many of its adherents, decreasing the incidence and improving the clinical course of CMV retinitis. However, a recent phenomenon associated with HAART, immune recovery uveitis (IRU), has been noted with additional, often problematic ocular manifestations in many persons with CMV retinitis. Periodic ophthalmic examination is prudent for all persons with HIV and especially for people with AIDS. PMID- 11387804 TI - Battering and HIV in men who have sex with men: a critique and synthesis of the literature. AB - Recent empirical evidence demonstrates the relationship between HIV and battering victimization in men who have sex with men (MSM). Although there is an abundance of literature examining domestic violence among heterosexual couples, there is minimal empirical evidence regarding battering among same-sex male couples. To obtain an understanding of the current state of the science, this literature review provides a critique and synthesis of the identified battering and HIV literature in MSM. After an examination of the relationship between HIV and battering, the prevalence of battering in MSM from the literature is critically reviewed. To establish a framework for understanding battering in MSM, the identified correlates of battering for both the victim and the perpetrator are examined. This critique and synthesis concludes with implications for clinical nursing practice. PMID- 11387805 TI - Spirituality and HIV disease: an integrated perspective. AB - Spirituality is an important resource that individuals use to cope with a chronic illness such as HIV disease. Spirituality has both a religious and an existential component that share the concepts of meaning in life, hope, self-transcendence, and rituals. An integrated perspective utilizing these shared concepts is proposed to assist HIV-positive individuals in coping with the challenges of their disease. Nursing interventions include promoting hope, teaching, sharing information, and creating a sense of empowerment in people with HIV to address spiritual issues. The article concludes with a case study that emphasizes application of the integrated perspective of spirituality with an HIV-positive person. PMID- 11387806 TI - Assessment of hunger and appetite and their relationship to food intake in persons with HIV infection. AB - Assessments of hunger and/or appetite are common methods of screening for development of illness-related anorexia. There are limited data to determine whether these methods predict actual food intake in persons with HIV disease. Therefore, the authors examined the relationship between self-reported food intake and subjective ratings of hunger and appetite in 31 adults with HIV infection. Participants also indicated presence of additional factors that can decrease amount of food eaten. Subjective ratings of appetite and hunger correlated with each other but not with food intake. Twenty-four additional factors that can affect food intake were reported to be present. The most common were illness-related and factors such as eating with friends or family. These results indicate that measures of hunger and appetite are not sufficient to screen for decreased food intake. Additional factors that can affect food intake should also be included in a comprehensive assessment of adults with HIV infection. PMID- 11387807 TI - Measurement of fatigue in HIV-positive adults: reliability and validity of the Global Fatigue Index. AB - Fatigue is among the most common and distressing symptoms in patients with HIV/AIDS. Little is known about the clinical assessment of fatigue, especially in patients using highly active antiretroviral regimens. The purpose of this study was to evaluate the psychometric properties of the Global Fatigue Index (GFI) in a community-based sample of 209 patients with HIV/AIDS. The GFI is a measure that quantifies five dimensions of fatigue from the Multidimensional Assessment of Fatigue instrument into one score. To assess construct validity, the study included measures of depression, perceived stress, activities of daily living (ADLs), health behaviors, and clinical markers. Cronbach's alpha was calculated for internal consistency reliability, and factor analysis and bivariate correlations were conducted. The GFI was found to be easily self-administered, reliable, and a valid measure of overall fatigue burden in an HIV population. This instrument may be used by clinicians and researchers for assessing fatigue. PMID- 11387808 TI - Patients tell of their images, expectations, and experiences with physicians and nurses on an AIDS-designated unit. AB - Throughout the 1980s and 1990s, there were only a few studies addressing the relationship of health care professionals and patients with AIDS from the patient's viewpoint. This descriptive, qualitative study explored patients' perceptions and experiences with physicians and nurses on an AIDS-designated unit in New York City. The data reveal patients' physical and emotional images and expectations of physicians and nurses. In meeting their needs, patients expect physicians and nurses to attend to their physical needs as the priority but do not expect them to address their emotional and social needs. Although patients appreciate recognition of their spiritual needs, they attempt to avoid discussions about religion. Patients characterized "special" and "difficult" physicians and nurses. Patients emphasized the importance of establishing a caring relationship through the professionals' therapeutic use of self. With knowledge of what is valued by patients in their relationship with physicians and nurses, health professionals have an opportunity to reflect on their own caregiving experiences and learn ways of enriching and supporting those relationships with the intent of improving the quality of care. PMID- 11387809 TI - Evidence of hypertriglyceridemia in managing HIV patients on efavirenz. AB - Of the recently approved antiretroviral agents, efavirenz has become a popular medication for the treatment of HIV infection. The efficacy of an efavirenz-based regimen has allowed the use of a combination of this medication with other antiretroviral agents in all levels of HIV disease, resulting in decreased viral replication and positive clinical results. Efavirenz is also proposed as an acceptable agent to switch to from a protease inhibitor-based regimen to reduce the risk of metabolic complications. In addition, the favorable side-effect profile, diminished pill burden for clients, and daily dosing have contributed to its popularity. Primary care practitioners should be cognizant of the possible side-effect profile of antiretroviral therapy combinations and enter into a collaborative relationship with the infectious disease clinician to ensure safe and effective patient care. PMID- 11387810 TI - The HIV-positive nurse--implications for clinical practice. PMID- 11387812 TI - [What is your diagnosis? Boerhaave syndrome with pneumomediastinum/pleural empyema]. PMID- 11387813 TI - [Immunomodulation by nutritional therapy--wish or reality?]. PMID- 11387814 TI - [Budd-Chiari syndrome in polycystic kidney disease]. AB - We present a patient with adult polycystic kidney disease, who developed a Budd Chiari Syndrome due to compression of the hepatic venous outflow tract by liver cysts. In patients with adult polycystic kidney disease who present a fast ascites formation, it is of great importance to look for a Budd-Chiari Syndrome as a complication of liver cysts. PMID- 11387815 TI - [Chronic progressive polycythemia and thrombocytosis]. AB - The 73-year-old female patient presented with chronic progressive erythrocytosis diagnosed 5 years ago which now is accompanied by thrombocytosis. She complains about generalised itching, pain in distal limbs and shows marked plethora. Having excluded major causes of secondary erythrocytosis the diagnosis was polycythaemia vera with a typical constellation of symptoms, findings and course of disease. Under treatment with 5-Hydroxyurea and repeated phlebotomy thrombocyte count and haematocrit normalised and the patient recovered her usual vitality. Limb pain, itching and plethora disappeared. PMID- 11387816 TI - [Generalized lymphadenopathy in a 43-year-old patient]. PMID- 11387817 TI - [Threatening hemoptysis]. PMID- 11387818 TI - [Threatening hemoptysis. Results of treatment with arterial embolization]. AB - OBJECTIVE: Retrospective analysis the immediate and long-term efficacy of embolization of bronchial and systemic arteries in the treatment of threatening or relapsing hemoptysis. MATERIALS AND METHODS: During the study period 122 arterial embolizations were performed in patients with hemoptysis over 100 ml in 24 hours, relapsing hemoptysis and/or presence of vital risk factors. Embolization was performed with polyvinyl alcohol particles, spongostan and metallic spirals. Seventy patients were included in the study and 47 were excluded as they came from other institutions. RESULTS: Angiographic changes were observed in 100% of patients. The immediate clinical success, defined as hemoptysis control, was obtained in the 70 patients. The mean follow-up time was 21.2 +/- 16.3 months. Relapsing hemoptysis occurred in 17.1% of patients (12 patients), of which 5.7% (4 cases) occurred in the first seven days and in 11.4% (8 patients) after 6 months. In five patients (7.1%) a new embolization was performed and 6 were operated after embolization (5 of them with bronchiectasis). The remarkable complications derived from the procedure included self-limited paraparesis of the lower limbs and severe chest pain. CONCLUSIONS: The embolization of bronchial arteries is an efficient technique for the treatment of threatening hemoptysis and relapses, is associated with a low morbidity rate, and the late relapse is relatively common among patiets with bronchiectasis. PMID- 11387819 TI - [Prevalence of antiphospholipid syndrome in patients under 65 years of age with acute myocardial infarction]. AB - BACKGROUND: The antiphospholipid antibody syndrome (AAS), which is characterized by thromboembolic events and/or fetal loss and/or low platelet count associated with antiphospholipid antibodies, may evolve with acute myocardial infarction (AMI). The presence of AAS among young patients with AMI ranges from 14% to 21%, and this condition implies specific therapeutic attitudes as new thrombotic events may occur, according to some authors. MATERIALS AND METHODS: A prospective study was undertaken with 25 patients aged > or = 65 years with AMI that were admitted to our institution during one year who were compared with control patients with similar risk factors. IgG and IgM anticardiolipin antibodies (ACA) were measured in the first 24 hours since the onset of AMI symptoms and three months later. RESULTS: The follow-up ranged from three months to one year. Among patients, ACA positivity in the two measurements was higher (12%) than that observed in the control group (5%) (p = 0.36). ACA positivity on two occasions was not a risk factor for new thrombotic events. CONCLUSION: ACA positivity is higher among AMI patients (measured early and at three months) than among the general population although the presence of such antibodies does not increase the risk for new post-infarction thrombotic events. PMID- 11387821 TI - [Miliary tuberculosis associated with acute respiratory distress and pancytopenia in HIV-negative patient]. AB - The acute respiratory distress syndrome (ARDS) is a rare but serious complication of miliary tuberculosis with a mortality rate near 100% when associated with pancytopenia. This association has been rarely reported (eleven cases in a Medline search, 1966-1999). A case is here reported of an HIV-negative patient with miliary tuberculosis which presented as ARDS associated with pancytopenia. This case prompted us to review the literature and risk factors, diagnosis and treatment issues are discussed. In patients with predisposing factors miliary tuberculosis must be considered as a possible cause of ARDS. PMID- 11387820 TI - [Analysis of prognostic factors for mortality in bacteremia and fungemia at a university hospital. Experience of 10 years]. AB - OBJECTIVE: The objective of this study was to determine the prognostic factors in relationship to evolution to death of bacteremia-fungemia (BF) episodes occurred in 1986 and to compare them with the results obtained ten years later in 1996. PATIENTS AND METHODS: Prospective study of all BF episodes observed at Hospital Universitario La Princesa, Madrid, during the 1985-1986 and 1996-1997 periods. The same definitions were used for the two study periods. The univariate analysis of results was performed with the chi square test and variables with statistical significance with p < 0.10 in the multivariate analysis with the logistic regression model. RESULTS: A total of 984 episodes were analyzed. There was an increased incidence per 1,000 admissions from 23.58 to 28.44. A change in the relationship of nosocomial acquisition (55.5%-42.6%) to community-acquired episodes (44.1%-57.4%) and an increase in gram-positive organisms (39%-48.6%) compared with gram-negative organisms (53.4%-41.8%) was observed. The organisms recovered most frequently in both periods were Escherichia coli and coagulase negative Staphylococcus. An overall decrease of mortality rate from 26.2% down to 15.9% (OR: 4.52) was noted. Independent factors with poor prognosis in the first period included age over 60 years (OR: 4.52), underlying disease (OR: 2.79; more than one OR: 6.53), respiratory source (OR: 3.86), DIC (OR: 4.79), hypotension (OR: 3.19); as for the second period, the corresponding independent factors included age > 60 years (OR: 6.48), nosocomial acquisition (OR: 2.62), DIC (OR: 18.7), hypotension (OR: 3.07), and inadequate surgical treatment (OR: 7.61). CONCLUSIONS: In the last ten years the incidence of BF episodes has increased. In contrast, mortality rate has decreased. Factors with poor prognosis, including age > 60, DIC, and hypotension, still persist. PMID- 11387822 TI - [Diagnostic assessment in patients with fever syndrome after traveling to the tropics]. PMID- 11387823 TI - [Alcoholic myocardiopathy]. PMID- 11387824 TI - [Etiopathogenic diagnosis of chronic and recurrent pancreatitis]. PMID- 11387825 TI - [Treatment of Graves' orbital disease]. PMID- 11387826 TI - [Treatment of acute tubular necrosis]. PMID- 11387827 TI - [Treatment of viral hepatitis]. PMID- 11387829 TI - [A 40-year-old man with left knee pain and tumefaction]. PMID- 11387828 TI - [A 70-year-old woman with right calf pain and inflammation]. PMID- 11387830 TI - [Radiologic diagnosis]. PMID- 11387831 TI - [A 74-year-old woman with staphylococcal bacteremia and lumbar pain]. PMID- 11387832 TI - ["Frozen" shoulder in a young woman]. PMID- 11387833 TI - [Nevirapine and clinical withdrawal syndrome]. PMID- 11387834 TI - [Intensification of antiretroviral treatment and adhesion]. PMID- 11387835 TI - [Hospital readmission in internal medicine and average stay]. PMID- 11387836 TI - [Neoplastic patients with low-risk neutropenic fever induced by chemotherapy as potential beneficiaries in home hospitalization programs]. PMID- 11387837 TI - [Oxidation, between life and disease]. PMID- 11387838 TI - [Elderly population and campaigns for the prevention of hypercholesterolemia in Salamanca]. AB - OBJECTIVE: To analyze the cardiovascular risk profile of the persons older than 65 years of age who participated in different "Days on Cholesterol and Atherosclerosis" organized during 1999 in different towns of the province of Salamanca. MATERIAL AND METHODS: The volunteers of any age were subjected to a questionnaire and to several clinical and laboratory measurements. RESULTS: The age of 267 volunteers (40.8% males) attending the campaign was > or = 65 years. The mean values of the studied parameters were: age = 72.3 +/- 6.5 years; body mass index (BMI) = 26.8 +/- 4.5 kg/m2; systolic blood pressure (SBP) = 139.8 +/- 20.6 mmHg; dystolic blood pressure (DBP) = 80.0 +/- 10.1 mmHg; and total cholesterol (TC) = 226.2 +/- 44.9 mg/dl. The percentage of smoking habit was 5.2%. The figures of SBP showed positive significant correlation with those of age (p < 0.05) and DBP (p < 0.01). Among males, the age evolved inversely to the numbers of TC (p < 0.01). Globally, 203 volunteers (76.0%) had concentrations of TC > or = 200 mg/dl; 78 (29.2%) > or = 250 mg/dl; and 12 (4.5%) concentrations > or = 300 mg/dl. We detected blood pressure levels of SBP > or = 140 mmHg in 143 (53.6%) individuals; SBP > or = 180 mmHg in 11 (4.1%); DBP > or = 90 mmHg in 72 (27.0%); and DBP > or = 110 mmHg in 4 (1.5%). The BMI was > or = 30 kg/m2 in 54 (20.2%) people of the study. 90.8% of the volunteers with previous cardiovascular disease carried, at least, another not controlled cardiovascular risk factor; 24.1% had at least two risk factors; and 9.1% had, at least, three. In 38.5% they showed high levels of TC and DBP, simultaneously. CONCLUSIONS: Our data, together with the current evidences about the benefit of the control of cardiovascular risk factors in the elderly, suggest the necessity to establish strategies for a better control of the cardiovascular risk in this group of age in our province. PMID- 11387839 TI - [Epidemiologic study on tuberculosis in the health area of Santiago de Compostela in 1995, 1996, 1997, and 1998]. AB - BACKGROUND: We report the epidemiological characteristics of tuberculosis(TB) in the area of Santiago de Compostela (Spain) between 1995 and 1998. METHODS: Inclusion criteria were: 1) microbiological and/or pathological diagnosis of TB in any specimen, and 2) consistent recent medical history of TB with reading > 5 mm to 2 TU PPD tuberculin test 48-72 hours after injection, and adenosine deaminase in pleural effusion > 47 IU/ml. RESULTS: 1,150 patients were included (685 males and 465 females), with mean of age (X) 38.9 years (19.8 SD), range 3 months-88 years. The number of cases was 307 during 1995 and 1996, 302 in 1997 and 213 in 1998. The incidence rate (per 100,000 habitants) was 78.3 in 1995 and 1996, 79.8 in 1997 and 61.9 in 1998. The incidence rate of meningitis was 1.8 in 1995, 1.3 in 1996, 1.05 in 1997 and 0.8 in 1998, with no meningitis in children under 5 years. Cases in association with VIH were 4.2% in 1995, 3.3 in 1996, 5.4 in 1997 and 3.2 in 1998. More of the 50% of cases in both genders were between 15 and 40 years old with another peak over 65 years. The ratio men/woman was 1.8 in 1995 and 1.4 in 1996, 1997 and 1998. CONCLUSIONS: The incidence of tuberculosis has diminished during the last year, but is too early to know the real tendency. PMID- 11387840 TI - [Atypical manifestations of Whipple's disease]. AB - We report 2 new cases of Whipple's disease (WD) with especial characteristics. In one case, an asymptomatic leukocytosis was the first manifestation, and after therapy the patient developed a gastric adenocarcinoma. In the second, the clinic suspect we led to try antibiotic treatment in absent of histological manifestations. The clinical, analytical and histological characteristics of both patients are described. We propose that WD should be listed in the differential diagnosis of unknown leukocytosis. Moreover, we indicate the possibility to try empirical antibiotic therapy in long-term cases without histological confirmation. Finally, we suggest a possible role of WD in the development of cancer. PMID- 11387841 TI - [Hypersensitivity to allopurinol. Efficacy of a desensitizing protocol in 3 cases]. AB - Allopurinol is often prescribed for the treatment of hyperuricemia. It inhibits the uric acid production binding tightly to xanthine oxidase. Although it is generally well tolerated, an almost 10% prevalence of adverse reactions has been reported, particularly gastrointestinal and neurological effects. Some hypersensitivity syndromes have also been described (rash, vasculitis or exfoliative dermatitis). In these cases, if a substitute treatment is not available, a desensitization procedure to the drug must be considered. We present three patients with cutaneous hypersensitivity to allopurinol, two who developed urticaria and other one who had a fixed drug eruption. Skin test were all negatives with positive oral challenge test. An out- patient desensitization procedure to allopurinol was initiated, repeating the last tolerated doses for 4 or 5 days, and reaching maintenance therapeutic drug doses without any significant adverse effect (only one case of cutaneous pruritus). These experiences and the previously reported in the literature, show that the desensitization to allopurinol is a good therapeutic alternative in hypersensitivity reactions to the drug. PMID- 11387843 TI - [Lung involvement in tularemia]. AB - We present three cases of pneumonia by Francisella Tularensis recently diagnosed. We also review this disease with the literature. All the studied patients were adults; two of them had epidemiological antecedents because of being in contact with hares. They present a clinical-radiological symptoms compatible with the pneumonic case described in the literature. The diagnosis was realized through serology in two cases and hemocultive in the other one. All patients had a positive answer to the antibiotic treatment, two cases with gentamicine and the other one with macrolide. There are not references about the subject in the bibliographical research we have realized in Medline. We did not find information in the Spanish base (IME), perhaps that these were the first cases found in the Spanish literature. PMID- 11387842 TI - [Triple association: late-onset seronegative spondyloarthropathy, Paget's disease and giant cell arteritis]. AB - The diagnosis of several rheumatic diseases in one-self patient is frequent in practice. However, some associations are exceptional and their recognition is important for a correct treatment and prognostic. We present the case of an old patient in who we diagnose a late onset seronegative spondylarthropathy and a Paget's disease and that later it developed a giant cells arteritis. The association of ankylosing spondylitis with Paget's disease or with polymyalgia rheumatica, it has already been described in the literature, but our patient's triple association we have not found it. PMID- 11387844 TI - [Addisonian crisis as first manifestation of adrenal gland insufficiency in patient diagnosed with lung cancer]. AB - Adrenal insufficiency or Addison's disease is actually a rare illness associated with numerous pathologies. We describe the case of a fifty years old male with lung adenocarcinoma and metastasis in both adrenal glands, who was receiving chemotherapy with mytomicin, ifosfamide and cisplatin (MIC), and was diagnosed of adrenal insufficiency as a result of acute episode addisonian crisis. Many times, the clinic symptoms of adrenal insufficiency can go unnoticed due to its low specifity and to mixing up with other syndromes. Hypoadrenalism has been described in association with many tumours, specially with non-Hodgkin's lymphoma. It seems that there is a discordance between the number of patients with bilateral metastatic adrenal destruction and the documented cases of clinic insufficiency. Once the adrenal failure is suspected, the diagnosis and hormone replacement treatment are really easy. Addison's disease ethiologies are revised putting special emphasis on those related with cancer patients. PMID- 11387845 TI - [Reflex sympathetic dystrophy: psychological and psychopathologic features]. AB - It has traditionally been accepted that a predisposing psychological field exists with the appearance of the reflex sympathetic dystrophy. Nevertheless, there is no unanimous agreement in this interpretation, since surveys exist unanimous are in favour the reactive character of the psychological effect on reflex sympathetic dystrophy. In this study, we revise literature already published on the subject, present arguments on the psychological evaluation tests carried out to date and propose the possibility of using other test unanimous are easier to manage and to interpret. PMID- 11387846 TI - [Reflexions on the most appropriate approach with low-weight heparins for antithrombotic prophylaxis in hip and knee arthroplasty]. PMID- 11387847 TI - [Necrotizing fasciitis]. PMID- 11387848 TI - [Bronchiolitis obliterans organizing pneumonia and Coxiella burnetii]. PMID- 11387849 TI - [Early hepatopathy induced by ticlopidine]. PMID- 11387850 TI - [Acquired factor VIII deficiency]. PMID- 11387851 TI - [Significance of emergency care in the elderly patient]. AB - OBJECTIVES: To analyse the statistical and characteristics of the attendance to the elderly patient at the Emergency Department of a General Hospital. METHODS: Retrospective study of patients > or = 65 years assisted at the Emergency Department of the Hospital Clinico San Carlos (Madrid), during 1998, taking into consideration different parameters as age, sex, time of year, referred pathology and final destination; results were compared with data relating to the population under 65. RESULTS: Over 28.4% of the 129,586 subjects that attended the Emergency Department were elderly people. Those ones aged between 65-74 years demanded assistance more frequently, being women's requests higher, particularly among the eldest patients. Assistance requests were higher in winter time, being the most common reason for consultation medical and surgical general troubles (76.4%), followed by trauma emergencies (12.35%). Hospital admission's percentage of elderly people was three times higher than the non-elderly ones, needing more admissions men than women (29 vs 22.7%). CONCLUSIONS: The high statistical of urgent attendance to the elderly patient compared with younger people as well as the higher number of admissions due to this assistance, seem to make necessary the implantation of Geriatric Evaluation Units at the hospital's Emergency Departments. PMID- 11387852 TI - [Dental sepsis as cause of fever of unknown origin]. PMID- 11387853 TI - [Bucco-facial apraxia and anarthria, an exceptional debut form of corticobasal degeneration]. PMID- 11387854 TI - [Hypercoagulability and ischemic cardiopathy: potential interaction between antiphospholipid antibodies and cellular element implicated in hemostasis]. PMID- 11387855 TI - [Acute abdominal pain in the elderly]. PMID- 11387856 TI - [Anemia and functional incapacity at admission to a geriatric home]. AB - AIM: To ascertain the prevalence of anemia on admission to a nursing home, and to assess the relationship between the observed cases of anemia and the functional status of those subjects. METHODS: We studied 198 subjects: 82 men (41%) aged 75.8 +/- 8.8 years, and 116 women (59%) aged 78.2 +/- 8.3 years. Anemia was diagnosed according to the criteria of the World Health Organization. The classification as non-disabled, or physically or mentally disabled, was done according to the Scales of the Spanish Red Cross. RESULTS: Anemia was diagnosed in 36% of the males, being microcytic in 14%, normocytic in 83%, and macrocytic only in 3% of them. Among women there were 44% with anemia, which was microcytic in 16%, normocytic in 80%, and macrocytic in 4% of cases. The prevalence of anemia increased with age in both sexes. Among men, anemia was significantly associated (p = 0.013) with physical disability, whereas among women this association just fell off significance (p = 0.06). There was no association of anemia with mental disability. No association was found between serum concentrations of ferritin, vitamin B12, or folic acid, and the classification as non-disabled, or as physically or mentally disabled. CONCLUSIONS: Anemia is found in about 40% of the elderly on admission to our nursing home. Anemia is associated with older age and with physical disability, but not with mental disability. Whether anemia on admission entails a higher risk of disability onset during the stay in the nursing home remains to be elucidated. PMID- 11387857 TI - Enhancing personal wellness in counselor trainees using biofeedback: an exploratory study. AB - The purpose of this study was to explore whether biofeedback-assisted relaxation training could reduce stress-related symptoms and enhance personal well-being in a group of counselor trainees enrolled in a basic counseling skills course. Treatment participants received ten sessions of weekly biofeedback-assisted relaxation training, whereas the control participants received no intervention. The treatment group showed, significant improvements in several symptom areas measured by the Symptom Checklist 90-Revised: physical complaints (Somatization), personal inadequacy (Interpersonal Sensitivity), confused thinking or alienation or both (Psychoticism), and the overall number and severity of symptoms (Global Severity Index and Positive Symptom Total). The control participants showed no significant improvements in any symptom areas. Biofeedback-assisted relaxation was shown to be an effective stress-reducing intervention for counselor trainees, which resulted in a greater sense of personal well-being. PMID- 11387858 TI - The ethics of placebo controlled clinical research and applied psychophysiology. PMID- 11387859 TI - The ethical use of placebo controls in clinical research: the Declaration of Helsinki. AB - Medical ethicists have questioned the use of no-treatment (placebo and sham procedure) controlled studies of new therapies when safe and effective standard therapies are available for use as an active or "equivalence" control. Current ethical principles of conduct for biomedical research specifically prohibit designs that withhold or deny "the best proven diagnostic and therapeutic" treatment to any participant in a clinical study, including those individuals who consent to randomization into a control group. Studies of psychophysiological therapies are often criticized on the grounds they lack a placebo or sham treatment control group. This paper briefly reviews the history of the problem and discusses the ethical standards that govern human research as derived from the Nuremberg Code and the Declaration of Helsinki. An examination of the problem with regard to research involving EEG biofeedback therapy for Attention Deficit/Hyperactivity Disorder, Traumatic Brain Injury, and depression serves to highlight the issues. It is concluded that the active treatment control (treatment equivalence) design is most appropriate for those clinical studies examining disorders for which there is a known, effective treatment. Sham- or placebo-controlled studies are ethically acceptable for those disorders for which no effective treatment is available. PMID- 11387860 TI - Ethical research issues: going beyond the Declaration of Helsinki. AB - La Vaque and Rossiter made a strong, supported argument that it is unethical to use a "no treatment" control group in a research study if a known, effective treatment is available. Their argument is based on the supposition that the Declaration of Helsinki is the ethical world standard for research with humans. Their argument appears to be straightforward, but is not simple to apply. The issues are very complex, include issues not discussed in their argument, and can lead to a different conclusion as pointed out in this paper. The World Medical Association developed the Declaration of Helsinki as one of their official policies. The Declaration of Helsinki, however, is not accepted as the world ethical standard, as demonstrated by its lack of adoption by many professional associations or even by the United States Federal Government. Perhaps it is not mentioned because its ethical provisions are aspirational rather than mandatory as implied by La Vaque and Rossiter. Researchers and clinicians should also be aware of other ethical issues not directly discussed in the La Vaque and Rossiter paper. The Belmont Report is the basis for the ethical protection of human research subjects for at least 17 federal agencies and does not mention the Declaration of Helsinki. The Belmont Report mentions several ethical principles that form the basis for informed consent, risk/benefit assessment, confidentiality of data, subject selection, Institutional Review Boards, and other protections needed when doing research with human subjects. At least 2 of these core principles have direct implications to the discussion related to the use of placebo controls. The ethical principle of fidelity is also important in guiding research activities with human subjects. Researchers should be familiar with the La Vaque and Rossiter argument, the Belmont Report, and the federal policies developed to implement the provisions of that report, for example, Regulation 45 CFR 46. PMID- 11387861 TI - The neurophysiology of dissociation and chronic disease. AB - Dissociation as a clinical psychiatric condition has been defined primarily in terms of the fragmentation and splitting of the mind, and perception of the self and the body. Its clinical manifestations include altered perceptions and behavior, including derealization, depersonalization, distortions of perception of time, space, and body, and conversion hysteria. Using examples of animal models, and the clinical features of the whiplash syndrome, we have developed a model of dissociation linked to the phenomenon of freeze/immobility. Also employing current concepts of the psychobiology of posttraumatic stress disorder (PTSD), we propose a model of PTSD linked to cyclical autonomic dysfunction, triggered and maintained by the laboratory model of kindling, and perpetuated by increasingly profound dorsal vagal tone and endorphinergic reward systems. These physiologic events in turn contribute to the clinical state of dissociation. The resulting autonomic dysregulation is presented as the substrate for a diverse group of chronic diseases of unknown origin. PMID- 11387862 TI - Enhancing proper sitting position using a new sEMG protocol, the "minimax" procedure, with Boolean logic. AB - The modification of posture in children using sEMG is a new area that offers much potential to aid disabled children regain function. This article describes a new protocol, the "minimax" procedure, to aid children regain motor function in sitting. The protocol uses selective reinforcement to encourage recruiting/relaxing target sites (gluteus medius/maximus) in a predetermined way so as to encourage good form in sitting. Children often are unaware how to make changes in posture, and when they do succeed the progress is so slow that sustaining motivation is a challenge. This paper describes a procedure that helps the child recognize when they have achieved the correct posture, immediately rewards them for doing so, and generates data using Boolean operations to numerically substantiate the clinically observed changes in posture. PMID- 11387863 TI - [Persistent infiltrative pulmonary disease]. AB - Two patients, a woman aged 63 and a man aged 64 years, were admitted with pulmonary complaints and persistent infiltrative lung abnormalities as revealed in chest X-rays. Routine diagnostic analysis did not lead to a diagnosis. However, a pathological examination of biopsies acquired by means of video assisted thoracoscopic surgery (VATS), revealed bronchiolitis obliterans organising pneumonia (BOOP). In the first patient the BOOP manifested itself as a rapidly progressive disease with fever, pulmonary complaints and X-ray abnormalities. There was no response to standard antibiotic treatment. The other patient had suffered from rheumatoid arthritis for a considerable time and gradually developed BOOP. Both patients recovered following adequate therapy with high doses of oral corticosteroids. BOOP is a pathological-anatomical entity. It is a nonspecific excessive repair response to a variety of stimuli, such as infection, drugs, collagen vascular diseases, inflammatory disorders, transplantation, intoxication and irradiation. BOOP can also occur idiopathically. A high-resolution CT-scan is useful in distinguishing BOOP from interstitial pulmonary fibrosis and other interstitial lung diseases. An open lung biopsy is necessary for the diagnosis BOOP and is best performed by means of VATS. The treatment of BOOP consists of administering high doses of corticosteroids (prednisone 1 mg/kg/day) and if treated adequately, the prognosis is fairly good. Due to the extensive variety in aetiology, the specific diagnostic procedures and the good response to necessary treatment, BOOP should be considered in the differential diagnosis of patients with persistent infiltrative lung disease. PMID- 11387864 TI - [Physical diagnostics--peripheral arterial diseases]. AB - In a patient with a high a priori risk of peripheral vascular disease, the positive predictive value of an abnormal physical diagnostic examination is high. In patients with a low prior probability of peripheral vascular disease, the physical examination makes little contribution to the diagnosis or exclusion of arterial insufficiency. For this purpose the ankle-brachial systolic pressure index is preferable. Peripheral arterial disease is unlikely when this index is normal. However, a low ankle-brachial index necessitates further investigations to determine possible arterial insufficiency in the lower extremities. PMID- 11387865 TI - [Roaming through methodology. XXXII. False test results]. AB - The number of requests for diagnostic tests is rising. This leads to a higher chance of false test results. The false-negative proportion of a test is the proportion of negative test results among the diseased subjects. The false positive proportion is the proportion of positive test results among the healthy subjects. The calculation of the false-positive proportion is often incorrect. For example, instead of 1 minus the specificity it is calculated as 1 minus the positive predictive value. This can lead to incorrect decision-making with respect to the application of the test. Physicians must apply diagnostic tests in such a way that the risk of false test results is minimal. The patient should be aware that a perfectly conclusive diagnostic test is rare in medical practice, and should more often be informed of the implications of false-positive and false negative test results. PMID- 11387866 TI - [Diagnostic image (37). Bilateral ureteral reflux at the distal part in a bilaterally duplicated collecting system]. AB - A 5-year-old girl presented with repeated urinary tract infections. A voiding cystourethrogram showed bilateral reflux in the lower poles of a bilateral duplicated collecting system. PMID- 11387867 TI - [Inadequate pharmacotherapy in patients with obsessive-compulsive disorder]. AB - OBJECTIVE: To determine the adequacy of drug treatment for patients with obsessive compulsive disorder (OCD). DESIGN: Retrospective. METHOD: One hundred and fifty outpatients with OCD, admitted at the University Medical Centre in Utrecht, the Netherlands, were evaluated for severity as measured with the 'Yale Brown obsessive compulsive scale' (Y-BOCS). RESULTS: At the intake, 35% of the patients used no medication, 58% used an antidepressant, less than 1% an antipsychotic and 6% a benzodiazepine. The average active dose was taken by 12% of the patients, 5% took the maximum dosage and 41% too low a dosage. Consequently, 6 out of the 10 patients used the correct medication (antidepressant) and less than 20% used a sufficiently high dosage. Of the patients, 38% had never previously taken antidepressants, 31% had previously used one antidepressant, 17% two different antidepressants and 12% at least three different antidepressants; 13% had taken the antidepressant at the highest dosage, 18% at the average active dosage and 31% at too low (thus ineffective) a dosage. This means that from a pharmacotherapeutic viewpoint not more than 1 in 8 patients had had an adequate treatment history. CONCLUSION: The results of this study show that only 1 in 8 OCD patients were treated appropriately. PMID- 11387868 TI - [Attempted suicide with sustained release diltiazem]. AB - A 15-year-old woman was admitted to the intensive care unit after intentional auto-intoxication with 10 tablets of 200 mg diltiazem sustained release. She developed hypotension and became oliguric. This was followed by metabolic acidosis. Maximum plasma lactate concentration was 10 mmol/l and the highest measured plasma diltiazem level was 500 micrograms/l. The occurrence of pulmonary oedema due to adult respiratory distress syndrome (ARDS) was striking and the patient required mechanical ventilation for a period of three days as a result of this. After five days in the intensive care unit, she was transferred to a psychiatric hospital in a good physical condition. Sustained release diltiazem intoxications have previously been described on five occasions. PMID- 11387869 TI - [Visual impairment due to optic neuropathy in 2 patients on amiodarone therapy, i.e. ethambutol and isoniazide]. AB - Two patients, a 69-year-old man and a 49-year-old woman, developed a bilateral decrease in visual acuity, the male patient within two months of starting to use amiodarone (Cordarone) and the female patient within ten months of starting to use ethambutol (Myambutol) and isoniazid. Optic neuropathy occurred in both patients and this was probably caused by the medications. After the medication had been withdrawn, the male patient's vision remained poor (1/300); 6 months later the female patient's vision had improved to 0.8 and 1.0. Before prescribing oculotoxic medications, physicians should know about the patient's visual acuity and any ocular diseases the patient may have. The patient must also be advised to contact their doctor immediately if visual complaints occur. Before and during treatment with ethambutol, ophthalmological examinations should be carried out regularly. In the event of complaints, iatrogenic optic neuropathy must be distinguished from an ischaemic optic neuropathy. The latter is usually associated with an acute unilateral decrease in visual acuity. Withdrawal of the medication is indicated for neuropathy. However, although the vision may improve it will not be completely restored. PMID- 11387870 TI - Morphological changes in garlic (Allium sativum L.) microbulblets during dormancy and sprouting as related to peroxidase activity and gibberellin A3 content. AB - The aim of this work was to study the physiological mechanisms of dormancy and sprouting during post-harvest of garlic (Allium sativum L.) microbulblets produced by meristem culture of garlic seed cloves. The morphological changes occurring in garlic microbulblets were assessed from harvest till sprouting in relation with peroxidase activity and levels of gibberellins. Also the effect of a cold treatment (30 days at 4 degrees C) given 30 days after harvest was studied. The results showed that during the state of dormancy in garlic microbulblets formation of the leaf primordia and vascular differentiation of the storage leaf occurred, while increases of peroxidase activity and low levels of GA3 (the only active gibberellin identified) were found. At the end of dormancy the sprouting channel was formed, vascular differentiation established, and peaks of soluble peroxidase activity as well as of GA3 were observed. At day 90 post harvest, garlic microbulblets showed physiologically mature and able to sprout. Further on, bud expansion and decrease of GA3 levels characterized sprouting of the microbulblets. The cold treatment enhanced GA3 levels and anticipated the sprouting process. PMID- 11387871 TI - Increase of tissue lipid hydroperoxides as determination of oxidative stress. AB - Increased levels of lipid hydroperoxides (LOOH) are frequently associated with the oxidative mechanisms involved in physiological states as ageing and with serious pathological conditions. In the present work the physiological and the CCl4-induced lipid hydroperoxides levels in mice liver and kidney were determined. The analysis of LOOH tissue levels was performed through the oxidation of 1-napthyldiphenylphosphine (NDPP) into its oxide (ONDPP) and further quantification by high pressure liquid chromatography at 292 nm UV detection. The physiological level of lipid hydroperoxides levels was higher in the kidney (245 +/- 8 nmol LOOH/g of tissue) than in liver (164 +/- 5 nmol of LOOH/g tissue). After a single administration of CCl4 (0.25 ml/g) tissue LOOH reached a maximum level after 15 min (416 +/- 21 nmol/g kidney and 303 +/- 6 nmol/g liver) and then slowly decreased. LOOH levels in liver afforded an early indicator (15 min) of oxidative damage. LOOH levels in kidney remained significatively increased up to 60 min post administration. The described HPLC assay is a useful, simple and sensitive method to detect cellular oxidative stress and damage. PMID- 11387872 TI - Comparative morphologic placental types in Dasypodidae (Chaetophractus villosus, Cabassous chacoensis, Tolypeutes matacus and Dasypus hybridus). AB - Information about the morphology of placentas in armadillos is scarce, except for D. novemcinctus. A comparative study of morphologic placental types in armadillos is important in order to have a comprehensive view of the peculiar reproductive physiology in this family. The aim of this paper is to perform a comparative analysis of the morphological features of the placenta in Chaetophractus villosus, Cabassous chacoensis, Tolypeutes matacus and Dasypus hybridus in order to classify them in accordance with Grosser (1909). The placentas were studied macroscopically and histologically (light microscopy in 1 micron thick sections and electron microscopy for fine structure). The macroscopic study in the 4 studied species showed a similar pear-shaped placenta homogeneously villosus in almost all the surface. The histological analysis showed that the 4 studied species had a hemochorial type of placenta. This type of placenta was also found in D. novemcinctus (Dasypodidae), but it is different from those described for other xenarthrans. Hemochorial types of placenta have also been described in more modern mammals. Despite the many primitive features of the armadillos and the different anatomical and physiological features between the genuses of dasypodids, all the studied species share this structural type of placenta. PMID- 11387873 TI - Immunohistochemical detection of a very high density lipoprotein (VHDL) in ovarian follicles of Triatoma infestans. AB - The ability of Triatoma infestans ovarian follicles to synthesize a very high density lipoprotein (VHDL) has been examined by immunohistochemical methods. This kind of lipoprotein can be envisaged as a storage hexameric protein present in the hemolymph of some insect species. VHDL immunoreactivity is observed in oocytes at different stages of maturation. The antigen is present in the oocyte cytoplasm as well as in the follicular epithelial cells. The immunopositive reaction in the apical surface of follicle cells suggests both a VHDL synthesis and a secretion process. Furthermore, VHDL seems to be stored into oocyte in yolk granules. On the contrary, no immunopositive reaction is observed in the intracellular spaces between follicle cells, suggesting that VHDL is not incorporated from hemolymph into the oocyte. PMID- 11387874 TI - The sesquiterpene lactone dehydroleucodine reversibly inhibits Allium cepa L. root growth. AB - Here, we prove that dehydroleucodine, a sesquiterpene lactone, at low concentrations (25-100 microM) slowed down the Allium cepa L root growth by 22 70% respectively neither affecting cell viability nor cell size. Removal of the drug after 24 h incubation restored the normal growth rate of the roots. Higher concentrations (200 microM) of dehydroleucodine were deleterious for the roots. As cell size did not change, it is most likely that dehydroleucodine affected some event of cell division cycle making it longer. Thus, dehydroleucodine could be a useful tool to slow down cell proliferation. PMID- 11387875 TI - Immunocytochemical localization of different cell types in the adenohypophysis of the cichlid fish Cichlasoma dimerus (Heckel, 1840). AB - The adenohypophysis of the cichlid fish Cichlasoma dimerus was studied using the avidin-biotin-peroxidase method with antisera raised against piscine pituitary hormones and heterologous antisera against mammalian pituitary hormones. Antiserum raised against rabbit ACTH recognized a group of cells bordering the neurohypophysis (NH) in the rostral pars distalis (RPD). Anti-chum salmon prolactin (PRL) identified a compact group of cells in the periphery of the RPD. Gonadotropin II (GTH II), thyrotropin (TSH) and growth hormone (GH)-ir cells were localized in the proximal pars distalis. Ir-GTH II cells were also observed in the dorsal area of the pars intermedia (PI). Ir-GTH I cells could not be identified using anti-chum salmon GTH I, this may be due either to a failure of the antisera to recognize the gonadotropin or to a low expression of the hormone in adults of this species. PAS positive cells from the PI bound specifically with three different antisera raised against somatolactin (SL) of four different fish species. These cells surrounded deep branches of the NH in the PI. PMID- 11387876 TI - Effect of dyskinetoplastic agents on ultrastructure and oxidative phosphorylation in Crithidia fasciculata. AB - Ethidium bromide (EB) is an intercalating agent which binds specifically to the kinetoplast (mitochondrial) DNA (kDNA) of trypanosomatids. Accordingly, EB inhibits DNA replication, thus inducing dyskinetoplasty. Since in eukariotic organisms mitochondrial DNA encodes the genetic information for cytochromes b, aa3 and F0F1 ATPase, it seemed of interest to establish whether a similar effect occurs in Crithidia fasciculata, a trypanosomatid used for assay of potential trypanocidal drugs. Culturing of C. fasciculata in the presence of EB inhibited growth and induced dyskinetoplasty, as confirmed by electron microscopy. The kinetoplast of EB-cultured crithidia lost its characteristic arc shape, it was misplaced in the cell cytoplasm its matrix structure and membrane differentiation were specifically modified. Dyskinetoplasty decreased crithidia respiration and oxidative phosphorylation, as indicated by the lower ATP level, ATP/ADP ratio and adenylate energy charge. The interference of EB with kinetoplastic constituents synthesis was confirmed by the lack of action of EB on crithidia in the stationary phase of growth, that ruled out direct inhibition of oxidative phosphorylation enzymes. The lipophilic o-naphthoquinone beta-lapachone produced structural alterations in kinetoplast membranes, that correlated with inhibition of oxidative phosphorylation. These latter effects involved free radicals since they were prevented by free radical scavengers. PMID- 11387877 TI - Behaviour of microtubules in cells infected with Toxoplasma gondii. AB - Toxoplasma gondii proliferates within the parasitophorous vacuole of the host cell. Simultaneously with parasite division and vacuolar development, lipids traffic and change in the spatial distribution of organelles of the host cell cytoplasm occur. Using fluorescence microscopy, and antibodies recognizing tubulin, we showed that microtubules change their distribution during host cell infection by tachyzoites of T. gondii. In addition, transmission electron microscopy of thin sections and replicas of partially extracted cells showed that host cell microtubules concentrate around the parasitophorous vacuole. Such microtubules distribution was evident in early infection times and was more prominent after 24 h of infection, when parasitophorous vacuole was completely surrounded by microtubules. However, the meshwork microtubule filaments became slack or absent after 72 h of infection of host cell. Colchicine and taxol treatment altered the shape of the parasitophorous vacuole containing tachyzoites. These observations suggest a close association between microtubules and intravacuolar development of parasites. PMID- 11387878 TI - [Ovarian apoptosis]. AB - Apoptosis, or programmed cell-dead is one of the main mechanisms in the process of tissular loss. This is an active process of cellular depletion that participates in a direct manner in tissular homeostasis during the life span. This apoptotic process is a key event on the pathology of tumors development. The nature of apoptosis is genetic, but it is triggered by external factors. The caspases are been mentioned as responsible of the specific cellular lost, which is the final outcome. The equilibrium between proliferation and cellular death is determined by a balance of survival factors and death promoters such as genic regulators, hormones, cytokines and growth factors among others. In the ovary, apoptosis regulates follicular proliferation and differentiation. During embryogenesis the magnitude of the population of oogonias is mediated by apoptosis. During the ovarian cycle, apoptosis participates also in the follicular atresia process. It is probable that apoptosis is participating also directly in the accelerated follicular atresia. This mechanism has been hypothesized to be involved in the pathophysiology of premature ovarian failure. The role of apoptosis is some reproductive pathological events such as chronic anovulation, low ovarian reserve and early ovarian dysfunction is still not known. The knowledge of the role of apoptosis in such pathological conditions will contribute to the understanding of the ovarian physiology and will permit to intervene early during the natural history of the disease. PMID- 11387879 TI - [Most of the complaints in gynecology and obstetrics care are generated by perceptions stemming from unavoidable results]. AB - The annual rises in the cost of claims suffered by some countries had led to increases in: costs of the attention; malpractice premiums; health personnel stress level; risks for the patients with difficult problems, and lack of opportune attention. The intricate interaction between clinical state, responses variability and medical procedures flaws makes impossible stop unavoidable outcomes (UO). Though UO are not derived from negligence or inability, patients and relatives can see it as a malpractice result. OBJECTIVE: To determine the proportion of complaints generated by UO related perceptions (UORP) and their distribution in obstetric and gynecologic stages of care. MATERIAL AND METHODS: A search for claims derived from gynecologic or obstetric care was undertaken in the Medical Arbitrition National Commission (CONAMED) database, those presented between Jun. 1, 1996, and Nov. 30, 1998, were chosen. Some claims were derived directly from UORP (D-UORP claims), others were derived indirectly (UORP clamis); claims were grouped for stage of the attention in which they were originated; non D-UORP claims were grouped also by motives. RESULTS: In 625 claims (98.6% of total) was possible obtain precise information as required for the study. 79% of complaints were derived from UORP; 17% from perceptions generated for medical activities unrelated to UO (MAU-UO claims) and 4% by perception originated from care system (CS claims). 46% of complaints concerned surgical treatment, 27% medical treatment, 14% initial study, 2.1% delivery, in 10% a specific stage of the attention was not mentioned. The motives of the non D-UORP complaints were: 1) I-UORP claims; a) opportunity of the action, when the patient referred delay or inopportuneness as cause of the complaint (17%, overall); b) Professional quality, when questioned de outcomes or medical criteria (23%); 2) MAU-UO claims: a) Professional capacity when patient questions ability, or complaint of error in diagnosis (failure to diagnose or incorrect diagnosis), wrong removal, unnecessary treatment or other actions outside of the norm (13%); b) Improper information (3%); c) Inappropriate manner (0.8%); 3) CS claims: a) Resources (1%); b) Accessibility (2.9%). COMMENTS: The damage risk attributable to medical negligence is very low among patients who had received obstetrical or gynecological care, most of the complaints are UORP generated. In obstetrical care--from 1997 national statistics and described results--a benefit was procured to 1,705,161 persons and were produced 17 possible wrong attentions by professional capacity, five by improper information and one by inappropriate manner; the rest, 123 complaints, were derived from perceptions induced by UO. Undoubtedly there is low complaint registration, however, even at worst the wrong attention effect is lower than the impact attained if the possibility of economic benefit search through medical demands is not totally canceled. The damage risk derived from cost of claims is greater, as suffered on many countries. Medical associations in countries without professional liability problems must accomplish negotiations so that the legislation does not establish compensations by medical care results, the request may be based in: a) the deleterious consequences derived from cost of claims; and, b) the low probability of affecting the patients interests, in Mexico complaints derived from perceptions about the professional capacity are one in 103,022 obstetrical care attentions, the improper information derived one in 316,989 and the inappropriate manner generated one in 1,373,620. These organizations and other health related could assume the commitment to abate the foreseeable complaints and to do research on feasible forms to forecast UO, technically this must be long-range and though international collaboration. PMID- 11387880 TI - [Embryo transfer: a critical step in assisted reproduction]. AB - The advances and development of new procedures and techniques related to embryo development, and its final outcome, the embryo transfer; have increased significantly the pregnancy rate through this reproductive assisted technique. The embryo transfer is probably the last part in in-vitro fertilization programs and the success should be determined through the technical approach and procedure's characteristics. The use and standardization of the current techniques during the embryo transfer process allow increasing the success in pregnancy and implantation rates in a short period of time in the current Assisted Reproduction Programs. PMID- 11387881 TI - [Association between gestational age, weight, and Apgar score in newborns extracted in pelvic presentation]. AB - The analysis for the present study was derived from 343 singleton pregnancies with a breech presentation. The variables associated with Apgar score at 5 minutes were, gestational age and birth weight. This association was observed in all patients and when they were analyzed separately according to route of delivery. The intrapartum and early neonatal mortality was absent in both groups. The perinatal outcome among the vaginally delivered frank breech infants at term was not statistically different compared with the group delivered by caesarean section. PMID- 11387882 TI - [Migraine in pregnancy]. AB - Migraine is a very common disturbance in women. frequency appears up to 18%, higher prevalence is between 20-50 years old and hormonal changes affect it along reproductive era. Physicians are concerned when migraine occurs or worse on pregnancy because of the potential effects of therapeutic measures on pregnancy or even in the child. OBJECTIVE: A review about definition, classification and theories to explain migraine, its effect on pregnancy and to set guidelines for the treatment during pregnancy and lactation. There are two theories to explain migraine: vascular and neurogenic. Pregnancy as a modifier of migraine features is a common belief. PMID- 11387883 TI - [Contribution to the classic technique of continuous caudal analgesia application. 1946]. PMID- 11387884 TI - How hospital's security combines personal service, advanced electronics. PMID- 11387885 TI - An interview with: Alan Lynch on establishing a chemical decontamination plan for your hospital. PMID- 11387886 TI - Hospital reorganizes security to provide 24/7 security/safety presence. PMID- 11387887 TI - The Oklahoma ice storm, a Y2K disaster that arrived one year later--how two rural hospitals coped and what they learned. AB - An unexpected ice storm last December in southeastern Oklahoma cut off power and water for days in the area and punched holes in the disaster plans of the two hospitals most affected. In this report, hospital officials describe how they fared and tell what they will do in the future to be better prepared for such a worst-case scenario. PMID- 11387888 TI - Proximity system added to hospital's 'progressive' security approach. PMID- 11387889 TI - Parking security update. AB - Protecting patients, employees, and their vehicles in hospital parking facilities continues to be a major assignment for hospital security. The following brief reports provide information on improving security both for planned and existing facilities and for more efficient operations. PMID- 11387890 TI - The next iteration of managed care? Panel discussion. AB - What did our five panelists predict for the next iteration of managed care? What do they see happening now? What will the health care industry look like in ten years? Employers will shift an increasing share of rising medical and prescription drug costs to consumers through higher co-pays and defined contributions. Some providers may market their services to attract these consumers. Tightly managed HMOs will provide access to lower-income consumers. So for, e-health is helping managed care organizations cut administrative costs, but not medical expenses. Genetic testing and therapies will not be commercially available for ten to 15 years. Based on demographic trends and labor shortages, demand is likely to exceed the capacity of the health care system within the next three decades. Our five experts caution that managed care is as unpredictable and chaotic as ever, but they do their best to tell you how they think it'll all play out in the near term. PMID- 11387891 TI - Physicians lead the way at America's top hospitals. AB - The 100 Top hospitals are selected annually based on seven critical parameters for each of the 6,200-plus U.S. hospitals with 25 or more beds. They include the previous year's risk-adjusted patient mortality and complication rates, severity adjusted average patient lengths of stay, expenses, profitability, proportional outpatient revenue, and asset turnover ratio (a measure of facility and technological pace-keeping ability). The winners are selected from five comparable size groupings--small, medium, large community, teaching, and large academic hospitals. Conspicuous among the winners at every level are physician led organizations. Even in the majority of hospitals headed by non-physician administrators, however, the managerial capabilities of medical directors are the key to success. The most common characteristic of these award-winning hospitals is that the leadership is working together and communicating the institution's goals effectively to all levels of the organization. PMID- 11387892 TI - 12 ways to be a better leader. AB - What are the prerequisites for leading successfully in today's turbulent health care environment? An entrepreneurial attitude, an emphasis on people management, and the ability to lead and manage change. This article offers a dozen suggestions for fostering adaptability and helping sustain the organization's purpose/mission: (1) Celebrate the workforce; (2) remove barriers; (3) allow people to take risks; (4) stop managing other people's problems; (5) prioritize organizational values; (6) stop managing for consensus; (7) segment your marketplace; (8) understand who the competition really is; (9) establish new relationships; (10) forget about employee satisfaction; (11) stop budgeting departmentally; and (12) beware of sacred cows. Courage is perhaps the single greatest attribute of transformational leadership. PMID- 11387893 TI - Informal leadership support: an often overlooked competitive advantage. AB - As environmental pressures mount, the advantage of using the same strategies and tactics employed by competitors continues to shrink. An alternative is adapting and applying answers successfully employed in other industries to health care organizations. Working with informal influence leaders to share your change management efforts represents one such example. Informal influence leaders offer an often-overlooked source of competitive advantage--they have already earned credibility and respect from others, who regularly look to them for guidance. When sharing their views, they significantly influence the acceptance or rejection of new initiatives. Influence leaders reach into every conversation, every meeting, and every decision made in an organization. The important question is whether they will exert their leadership in support or in opposition to changes you propose. By identifying influence leaders and inviting them to join a group to discuss change initiatives, physician executives can create a positive force for change. PMID- 11387894 TI - The 7 habits of highly effective medical directors. AB - Like the well-known 7 Habits of Highly Successful People, the seven steps for successful medical management outlined in this article offer an inspirational guide for physician leadership in today's chaotic health care arena. Setting a vision, communicating the vision, and leading employees to realize the vision may sound like the simple characteristics of any leader. True leaders, however, must be prepared to delve deeply into their health care organizations. They must understand the inner workings of their committees and develop positive relationships with the staff. They must provide the technical tools necessary for the staff to work toward the vision, and understand the measured steps that managers must take along the path to achieving success for the entire organization. PMID- 11387895 TI - What skills do physician leaders need now and in the future? AB - The role of the physician leader is moving beyond traditional medical staff issues. A recent national survey of physician leaders shows a growing need for education on specific technical, leadership, and practical skills. The results reveal the medical leadership skills that physician executives consider important today, and provide a window to the future about the skills that will be important tomorrow. Physicians say they need training now in quality assurance, clinical benchmarking, decision-making, and strategic planning. And when they gaze into the future and see the rapid changes throughout health care, they say they'll need more training in communication, organizational change, effective listening, and systems thinking. PMID- 11387896 TI - Partnerships in health care: creating a strong value chain. AB - The health care climate is one of stormy relations between various entities. Employers, managed care organizations, hospitals, and physicians battle over premiums, inpatient rates, fee schedules, and percent of premium dollars. Patients are angry at health plans over problems with access, choice, and quality of care. Employers dicker with managed care organizations over prices, benefits, and access. Hospitals struggle to maintain operations, as occupancy rates decline and the shift to ambulatory care continues. Physicians strive to assure their patients get quality care while they try to maintain stable incomes. Businesses, faced with similar challenges in the competitive marketplace, have formed partnerships for mutual benefit. Successful partnerships are based upon trust and the concept of "win-win." Communication, ongoing evaluation, long-term relations, and shared values are also essential. In Japan, the keiretsu contains the elements of a bonafide partnership. Examples in U.S. businesses abound. In health care, partnerships will improve quality and access. When health care purchasers and providers link together, these partnerships create a new value chain that has patients as the focal point. PMID- 11387897 TI - Got curiosity? AB - The newest scientific models of decision-making suggest that the way we actually decide to do something is different from the story we put on it later. Organizations think that way, too. The real process of decision-making is hidden. Management is complex--and a key tool is curiosity. A critical question would be: "What experience are we creating?" When you get curious, separate what you notice from the story you tell yourself about it--even if the story is true. Look for the meaningful experience: your own, your employees', your customers', your physicians'. The essential act of management is to notice, to not look away. PMID- 11387898 TI - Chance encounters. AB - How can physician executives change their thinking and expand their understanding? Chance encounters offer a way to draw on others' unique experiences and patterns of thought unknown to us and have our usual way of thinking challenged and sometimes shattered--one of life's most powerful experiences. After reflecting on three optimal personal learning experiences from a recent conference, the author determines that the only common thread is the interdisciplinary thoughts expressed by a passionately involved individual whose approach to the problem is completely different from his own. These conferences offer the inquisitive physician entrepreneur the opportunity to hear how venture capitalists, population geneticists, business development experts, biotech CEOs, and big pharma scientists view the potential of the sequencing of the human genome to change medicine. Being able to see the world in a new way, the author concludes, is tied to the essence of entrepreneurship and the essence of joyful living. PMID- 11387899 TI - How to make the pitch for a part-time workload. AB - Are you planning on moving from full-time to part-time hours? You'll need a game plan to negotiate what you want, including establishing a timeline and agreeing on productivity expectations. If you can agree on a reasonable timeframe that doesn't inconvenience anyone or endanger important results or relationships, you have a high probability of getting your boss' okay. If you and the boss can't agree on what you must produce, don't consider part-time work unless you thrive on combat. Once you negotiate your new schedule, consider the issue of managing co-worker resentment. Here are the best hints for keeping co-worker envy and resentment at a manageable level: Don't be secretive; keep a low profile; attend all office frolics; and ask for a trial period. PMID- 11387900 TI - Changing what goes on in your head: how to stop "ain't it awful?". AB - How can you change your negative thinking? This column describes a process that, on the surface, seems too simplistic to be beneficial, but that works: choose a few good words to repeat to yourself constantly, progress to better thoughts, and then improve what you say to others. If you want to be more satisfied with your work life and your personal life, you must change the internal dialogue in your head. If you have some version of negative internal chatter, you need to substitute positive statements. You need to say something different from what you have been saying every spare minute of the day. You must say it even if it is the biggest lie you have ever heard yourself think. You must say it for days or weeks before you notice a difference in your attitude, relationships, and health. Eventually, you will notice you feel better and people are behaving better. PMID- 11387901 TI - The right paperwork for the right audiences. AB - Physician executives need to exercise considerable discretion, care, and judgment when they write about their professional accomplishments in the form of a resume or curriculum vitae (CV). This paperwork is intensely personal. It must be a true reflection of you, your achievements and goals. Others read it to learn more about you. Those you fail to convince, you're likely to lose--along with your chance to meet and charm them in a personal interview. A physician executive's thoughtfully prepared resume and CV can result in him or her being offered terrific opportunities for career growth. The paperwork is a self-constructed gateway through which you can properly approach hiring organizations--and how it reads is totally under your control. Some suggestions to consider in developing your resume or CV are: be succinct; don't overstate; use both documents; do your homework; and be ready for the next steps. PMID- 11387902 TI - Patient bill of rights 2001. AB - Breaking gridlock on managed care reform, a bipartisan coalition in Congress introduced the newest version of a patient bill of rights. Unlike last year's ill fated Norwood-Dingell bill, the Bipartisan Patient Protection Act of 2001 has strong bipartisan support; concern remains, however, on the provisions that allow patients to sue their managed care plan. The debate now focuses on the type of liability reform that Congress and the White House can agree on. If they are able to agree, a patient bill of rights may soon become law. PMID- 11387903 TI - Fast forward: a blueprint for the future from the Institute of Medicine. AB - A newly released report from the Institute of Medicine outlines an ambitious program for changing the direction of U.S. health care. Crossing the Quality Chasm: A New Health System for the 21st Century recommends switching health system priorities from predominantly acute care treatment to focusing on chronic medical conditions. The report also recognizes 15 conditions that it says should take priority for funding and support from all health care agencies. Evidence based medicine must be fostered and the entire fabric of medical care must become more patient-centered. The IOM report proposes six aims for our 21st Century health care system. The system we should strive for needs to be: (1) safe; (2) effective; (3) patient-centered; (4) timely; (5) efficient; and (6) equitable. This article looks at some of the IOM recommendations and analyzes their strengths and weaknesses. Ultimately, the report advocates an environmental restructuring of health care in the United States. PMID- 11387904 TI - Marijuana: federal smoke clears, a little. PMID- 11387905 TI - Remission of cold hemagglutinin disease induced by rituximab therapy. PMID- 11387906 TI - Escherichia coli infections and hemolytic-uremic syndrome. PMID- 11387907 TI - Escherichia coli infections and hemolytic-uremic syndrome. PMID- 11387908 TI - Pity the NHS. PMID- 11387909 TI - Prescription data. PMID- 11387910 TI - Prescription data. PMID- 11387911 TI - Prescription data. PMID- 11387912 TI - Prescription data. PMID- 11387913 TI - Prevention of influenza and pneumococcal pneumonia in Canadian long-term care facilities: how are we doing? AB - BACKGROUND: Influenza and pneumococcal pneumonia are serious health problems among elderly people and a major cause of death in long-term care facilities. We describe the results of serial surveys of vaccination coverage and influenza outbreak management in Canadian long-term care facilities over the last decade. METHODS: Cross-sectional surveys consisting of questionnaires mailed to all Canadian residential long-term care facilities for elderly people in 1991 and to a random sample of respondents in 1995 and 1999. RESULTS: The response rates were 83% (430/515) in 1995 and 75% (380/506) in 1999. In 1999 the mean reported rates of influenza vaccination were 83% among residents and 35% among staff, and the mean rate of pneumococcal vaccination among residents was 71%; all 3 rates were significantly higher than those in 1991. The rates were also higher in facilities with an infection control practitioner than in those without such an individual (88% v. 82% for influenza vaccination among residents [p < 0.001], 42% v. 35% for influenza vaccination among staff [p = 0.008] and 75% v. 63% for pneumococcal vaccination among residents [p < 0.001]). Obtaining consent for vaccination on admission to the facility was associated with higher influenza and pneumococcal vaccination rates among residents (p = 0.04 and p < 0.001 respectively). Facilities with higher influenza vaccination rates among residents and staff reported lower rates of influenza outbreaks (p = 0.08 and 0.03 respectively). Despite recommendations from the National Advisory Committee on Immunization, only 50% of the facilities had policies for amantadine prophylaxis during influenza A outbreaks. Amantadine was judged effective in controlling 76% of the influenza A outbreaks and was discontinued because of side effects in 3% of the residents. INTERPRETATION: Influenza and pneumococcal vaccination rates among residents and staff in Canadian long-term care facilities have increased over the last decade but remain suboptimal. Vaccination of residents and staff against influenza is associated with a reduced risk of influenza outbreaks. Amantadine is effective in controlling influenza outbreaks in long-term care facilities. PMID- 11387914 TI - Influenza vaccination in Alberta long-term care facilities. AB - BACKGROUND: Canada's National Advisory Committee on Immunization recommends that both staff and residents of long-term care facilities be vaccinated against influenza. This paper describes the influenza vaccination policies and programs, as well as vaccination rates, for staff and residents of long-term care institutions in Alberta. Such data have not previously been reported. METHODS: Data were collected by means of an anonymous mail survey (with 2 reminders) sent to Alberta nursing homes and auxiliary hospitals in spring 1999. RESULTS: Of 160 facilities providing long-term care during the study period, 136 responded to the survey (85%). Of these, only 85 provided data on staff vaccination rates, whereas 118 provided data on resident vaccination rates. For institutions reporting this information, the median proportion of staff vaccinated was 29.9% and the median proportion of residents vaccinated was 91.0%. Only 2 facilities reported that staff vaccination was mandatory; however, only one of these had a written policy consistent with the self-report period. Using a travelling vaccination cart, offering vaccination on night shift, and monitoring and providing feedback about staff vaccination rates were infrequently employed as elements of staff vaccination programs, although all were positively correlated with staff vaccination rates. Standing orders for resident vaccination were reported by only 84 facilities. Fourteen institutions required written consent for vaccination from the resident or a relative. Facility requirements for consent to vaccinate from the resident or a relative were significantly associated with mean vaccine coverage: 90.5% coverage for institutions requiring verbal consent, 86.5% coverage for institutions requiring written consent and 95.0% for institutions not requiring written or verbal consent. INTERPRETATION: Staff vaccination rates in Alberta long-term care facilities are unacceptably low. Changes in staff vaccination programs may improve the situation even in the absence of mandatory vaccination or work exclusion rules. Requirements for written consent for vaccination of residents of long-term care facilities may be a barrier to immunization. PMID- 11387915 TI - A case study of hospital closure and centralization of coronary revascularization procedures. AB - BACKGROUND: Despite nation-wide efforts to reduce health care costs through hospital closures and centralization of services, little is known about the impact of such actions. We conducted this study to determine the effect of a hospital closure in Calgary and the resultant centralization of coronary revascularization procedures from 2 facilities to a single location. METHODS: Administrative data were used to identify patients who underwent coronary artery bypass grafting (CABG), including those who had combined CABG and valve procedures, and patients who underwent percutaneous transluminal angioplasty (PTCA) in the Calgary Regional Health Authority from July 1994 to March 1998. This period represents the 21 months preceding and the 24 months following the March 1996 hospital closure. Measures, including mean number of discharges, length of hospital stay, burden of comorbidity and in-hospital death rates, were compared before and after the hospital closure for CABG and PTCA patients. Multivariate analyses were used to derive risk-adjustment models to control for sociodemographic variables and comorbidity. RESULTS: The number of patients undergoing CABG was higher in the year following than in the year preceding the hospital closure (51.6 per 100,000 before v. 67.3 per 100,000 after the closure); the same was true for the number of patients undergoing PTCA (129.8 v. 143.6 per 100,000). The burden of comorbidity was significantly higher after than before the closure, both for CABG patients (comorbidity index 1.3 before v. 1.5 after closure, p < 0.001) and for PTCA patients (comorbidity index 1.0 before v. 1.1 after, p = 0.04). After adjustment for comorbidity, the mean length of hospital stay was significantly lower after than before the closure for CABG patients (by 1.3 days) and for PTCA patients (by 1.0 days). The adjusted rates of death were slightly lower after than before the closure in the CABG group. The adjusted rates of death or CABG in the PTCA group did not differ significantly between the 2 periods. INTERPRETATION: Hospital closure and the centralization of coronary revascularization procedures in Calgary was associated with increased population rates of procedures being performed, on sicker patients, with shorter hospital stays, and, for CABG patients, a trend toward more favourable short-term outcomes. Our findings indicate that controversial changes to the structure of the health care system can occur without loss of efficiency and reduction in quality of care. PMID- 11387916 TI - Baseline staging tests in primary breast cancer: a practice guideline. AB - BACKGROUND: Breast cancer develops in over 7000 women each year in Ontario. These patients will all undergo some staging work-up at diagnosis. The Breast Cancer Disease Site Group of the Cancer Care Ontario Practice Guidelines Initiative reviewed the evidence and indications for routine bone scanning, liver ultrasonography and chest radiography in asymptomatic women who have undergone surgery for breast cancer. METHODS: A systematic review of the published literature was combined with a consensus interpretation of the evidence in the context of conventional practice. RESULTS: There were 11 studies of bone scanning reported between 1972 and 1980, involving a total of 1307 women; bone scans detected skeletal metastases in 6.8% of those with stage I breast cancer, 8.8% with stage II and 24.5% with stage III. A total of 5407 women participated in 9 studies of bone scanning reported between 1985 and 1995; in these studies, bone scans detected skeletal metastases in only 0.5% of women with stage I disease, 2.4% with stage II and 8.3% with stage III. Among 1625 women in 4 studies of liver ultrasonography reported between 1988 and 1993, hepatic metastases were detected in 0% of patients with stage I disease, 0.4% with stage II and 2.0% with stage III. Among 3884 patients in 2 studies of chest radiography published in 1988 and 1991, lung metastases were detected in 0.1% of those with stage I, 0.2% with stage II and 1.7% with stage III. False-positive rates ranged from 10% to 22% for bone scanning, 33% to 66% for liver ultrasonography and 0% to 23% for chest radiography. The false-negative rate for bone scanning was about 10%. RECOMMENDATIONS: The following recommendations apply to women with newly diagnosed breast cancer who have undergone surgical resection and who have no symptoms, physical signs or biochemical evidence of metastases. Routine bone scanning, liver ultrasonography and chest radiography are not indicated before surgery. In women with intraductal and pathological stage I tumours, routine bone scanning, liver ultrasonography and chest radiography are not indicated as part of baseline staging. In women who have pathological stage II tumours, a postoperative bone scan is recommended as part of baseline staging. Routine liver ultrasonography and chest radiography are not indicated in this group but could be considered for patients with 4 or more positive lymph nodes. In women with pathological stage III tumours, bone scanning, liver ultrasonography and chest radiography are recommended postoperatively as part of baseline staging. In women for whom treatment options are restricted to tamoxifen or hormone therapy, or for whom no further treatment is indicated because of age or other factors, routine bone scanning, liver ultrasonography and chest radiography are not indicated as part of baseline staging. PMID- 11387917 TI - Semmelweis revisited: the ethics of infection prevention among health care workers. PMID- 11387919 TI - Neonatal subgaleal hemorrhage: diagnosis and management. PMID- 11387918 TI - Lifestyle drugs: issues for debate. PMID- 11387920 TI - Chickenpox vaccination, not chickenpox, should be routine for Canadian children. PMID- 11387922 TI - Cuba trains American medical students--to work in US. PMID- 11387921 TI - Rheumatology: 13. Minimizing disability in patients with low-back pain. PMID- 11387923 TI - Prevalence of migraine headache in Canada. PMID- 11387924 TI - Acute care revolution on way in Saskatchewan? PMID- 11387925 TI - MDs' "no nukes" movement moves online. PMID- 11387926 TI - Canada ranks in middle in child-injury mortality rate, report indicates. PMID- 11387927 TI - Varicella vaccine in clinical practice. PMID- 11387928 TI - Illiteracy as a public health issue. PMID- 11387929 TI - [What is the role of angiotensin receptor antagonists in the treatment of cardiac failure?]. PMID- 11387930 TI - [Observations on the management of cardiac failure in ambulatory patients. Results of a survey of cardiologists]. AB - An enquiry was undertaken amongst 71 cardiologists in 1999 evaluating 600 ambulatory patients with cardiac failure (64% men; mean age 73 years; NYHA Class I: 9%, Class II: 52%, Class III: 33%, Class IV: 6%). Cardiac failure is commoner in men (81%) before the age of 70. After the age of 80, the tendency is inversed (53% of women). Fifty-two per cent of patients had a history of hospital admission for cardiac failure: 26% in the year before the enquiry (on average 13.1 days' hospital stay). On inclusion, the diagnosis was made 4.2 +/- 4.6 years previously and the patients had been followed up for an average of 3.04 +/- 3.3 years. The diagnosis of cardiac failure was made by the cardiologist (57%), the general practitioner (37%), or another category of physician (6%). Eight per cent of patients were treated with a single drug before inclusion, 22% had a bi therapy and 70% a polytherapy. The most frequently prescribed drugs were: diuretics (71%), angiotensin converting enzyme (ACE) inhibitors (54%) and digitalis (35%). The prescription of betablockers was only reported in 14% of the population. In the over-80's, only 45% of patients were prescribed ACE inhibitors. These results confirm that ambulatory patients with cardiac failure are mainly in NYHA Classes II and III, that the pathology concerns elderly patients and that it is the cause of recurrent hospital admission. The prescription of diuretics is the main therapy in all NYHA Classes. ACE inhibitors were only prescribed in 2/3 of patients and in less than half of patients over 80 years of age. The prescription of betablockers remains marginal. The reason for the underprescription of recommended drugs (International Recommendations) would justify further investigation. PMID- 11387932 TI - [Mid-term results of stentless bioprosthesis for aortic valve replacement. Experience in a series of 97 patients]. AB - Stentless bioprostheses have been described as valve substitutes of interest for aortic valvular replacement. We studied 97 consecutive patients with a mean age of 72.2 years (40-84) who underwent aortic valvular replacement with 80 Toronto SPV and 17 Freestyle prostheses. Operative mortality was 6.2. With a mean follow up of 19 +/- 10 months (1-46), 87.2% of the surviving 86 patients underwent an echocardiography performed by the same operator. Mean gradient was 10.9 +/- 3.6 mmHg (4.2-22.6) and effective orifice area was 1.8 +/- 0.5 cm2 (0.8-3.0) for the 75 controlled stentless valves. The best haemodynamic data were obtained with the 25 mm diameter prostheses. One asymptomatic partial dehiscence was observed during monitoring. None of the 15 detected aortic leaks was significant. We observed a significant reduction of the ventricular mass in 41 patients who had undergone pre- and postoperative evaluation (p < 0.0014). Overall survival was 86.8 +/- 4.4% at 2 years. Stentless bioprostheses offered satisfactory haemodynamic results in our series. They however require an implantation technique learning curve as well as a thorough knowledge of the aortic root anatomy and physiology. PMID- 11387931 TI - [Revision of the "Smoker's Paradox": smoking is not a good prognostic factor immediately after myocardial infarction]. AB - In order to determine the reasons for the low mortality after myocardial infarction in smokers compared with non-smokers (the smoker's paradox), the authors analysed the initial clinical data, the therapeutic interventions and hospital mortality in 790 consecutive patients (555 smokers, 235 non-smokers) admitted to hospital within 6 hours of the first symptoms of acute myocardial infarction and treated by intravenous thrombolytic agents and/or coronary angioplasty. Multivariate analysis with linear regression was used to identify the predictive factors of hospital mortality. The main differences between smokers and non-smokers were age (56 vs 67 years, p < 0.0001), gender (male, 90 vs 60%, p < 0.01), cardiogenic shock on admission (3 vs 8%, p < 0.01). TIMI 3 flow was obtained in the culprit artery in 84% of smokers and 79% of non-smokers (NS). Hospital mortality was 5% in the smoking population and 16% in non-smokers (p < 0.0001). In multivariate analysis, the variables of cardiogenic shock, age, gender and hypertension provided most of the prognostic information and tobacco consumption did not appear to have a protective effect. In patients admitted to hospital with acute myocardial infarction, identical incidences of early reperfusion are obtained in smokers and in non-smokers. However, mortality is higher in the non-smoking group due to more severe clinical characteristics on admission. Tobacco consumption is not a protective factor in the immediate period after acute myocardial infarction. PMID- 11387933 TI - [Three-dimensional modelling of thoracic aortic aneurysm. A case report]. AB - The anatomical forms of aneurysms are sometimes complex. Three-dimensional modelisation may be valuable in understanding the configuration and spatial orientation on one hand and also help in therapeutic decision making on the other. Two types of modelisation are reported with details of their methods and their respective values. PMID- 11387934 TI - [Endomyocardial fibrosis in Behcet's disease: a case report of a pseudo-tumoral form]. AB - Endomyocardial fibrosis is very rare in Behcet's disease. The authors report the case of a 28 year old patient with Behcet's disease complicated by a pseudo tumoral right ventricular formation on echocardiography. This misleading appearance suggested the diagnosis of cardiac thrombus or tumour and led to a surgical approach which revealed a fibrous moderator band suggesting endomyocardial fibrosis, confirmed by antomopathological analysis. Besides the originality of this case and the unusual pseudo-tumoral presentation, the authors underline the difficulties of establishing the diagnosis, despite the advances of medical imaging. The pseudo-tumoral intra-cardiac lesion in a suggestive clinical context (Behcet's disease) should raise suspicion of the diagnosis of endomyocardial fibrosis. PMID- 11387936 TI - [Coronary bypass surgery on the beating heart and surgery of an abdominal aortic aneurysm. Immediately sequential surgical treatments]. AB - The authors report sequential association during the same general anaesthetic of coronary bypass surgery on the beating heart and surgery of an abdominal aortic aneurysm. Two aorto-coronary bypass grafts were carried out without cardiopulmonary bypass using the two pediculated internal mammary arteries (without manipulation of the ascending aorta), followed, after closure of the chest and monitoring in the operating theatre for one hour, by reinstallation of the patient for treatment of an infra-renal abdominal aortic aneurysm by classical prosthetic implantation. The postoperative course was uncomplicated. Sequential management of coronary revascularisation without cardiopulmonary bypass and aortic aneurysmal lesions during the same anaesthetic provides an alternative to classical two-stage surgery in selected patients. PMID- 11387935 TI - [Simultaneous occlusion of the abdominal aorta and both internal carotid arteries as the presenting symptoms of left atrial myxoma]. AB - Myxoma is a benign tumour but which has redoubtable embolic complications. When situated in the left atrium, the emboli obstruct, in the majority of cases, the cerebral arteries, occasionally the visceral or coronary arteries, and, very rarely, the aorta. In this case, the authors report an atypical presentation with ischaemia of the lower half of the body, associated with pulmonary oedema and deep coma. The left atrial myxoma was responsible for complete and simultaneous obstruction of the internal carotid arteries and the infra-renal abdominal aorta. This report illustrates the fact that myxoma can be responsible for massive, life threatening, embolisation. PMID- 11387937 TI - [Mortality and quality of treatment in myocardial infarction]. PMID- 11387938 TI - [Cloning of rat beta-defensin rBD-1 cDNA]. AB - This is a study aimed at the molecular mechanisms of beta-defensins gene expression and it's gene transfer experiments for preventing mucosal infection. A rat rBD-1 cDNA was cloned. The total RNA was isolated from rat kidney. The cDNA fragment was amplified by RT-PCR with specific primers. The purified RT-PCR product was cloned in pGEM-T Easy vector. The results of restriction endonuclease pattern analysis of the recombinant plasmid, DNA sequencing, and aligning of the putative amino acid sequence with hBD-1 and mBD-1 demonstrated that rat beta defensin rBD-1 gene was cloned successfully. PMID- 11387939 TI - [Study on the reversal of cancer multidrug resistance by Chinese medicine Fw13 te41 in nude mice]. AB - We have reported that three reversal agents were sifted out from 32 Chinese galenicals through a series of cell culture tests. Among them, Fw13-te41 has the best effect of reversal cancer multidrug resistance (MDR) in vitro. In this study, the reversal action of Fw13-te41 in vivo was studied on the animal model of nude mice with human leukemia k562/ADR. Twenty SPF BALB/c-nu/nu nude mice with xenograft tumor were randomly divided into the control group (n = 6), VCR group [intraperitoneal (i.p.) VCR 250 micrograms/week, n = 5], VCR + Fw13-te41 group (i.p VCR 250 micrograms/week + Fw13-te41 0.2 ml/day, equivalent to crude drug 10 g/kg, n = 5), and Fw13-te41 group (i.p Fw13-te41 0.2 ml/day, equivalent to crude drug 10 g/kg, n = 4). After 18 days, the rate of tumor inhibition (RTI) of VCR group was 19.79%, but the RTI of VCR + Fw13-te41 group was as high as 86.95% (P < 0.05). There results demonstrate that the Chinese medicine Fw13-te41 has an evident reversal action of malignancy MDR in vitro and in vivo. PMID- 11387940 TI - [The establishment of a modified lateral fluid percussion model of brain injury in rat and the pertinent pathologic changes]. AB - For the purpose of studying the molecular mechanism of the traumatic brain injury, we have established a reproducible graded lateral fluid percussion model of experimental brain injury in the rat with a modified fluid percussion device. The device consists of a stainless steel cylindrical reservoir instead of the plexiglass reservoir, a steel reservoir filled with compressed gas instead of the pendulum for making more accurate percussion pressure, an apparatus for releasing the pressure immediately after the percussion, and a computer for recording and storing the percussion data. Pathologic examination demonstrated subdural hemorrhage, subarachnoid hemorrhage, and hemorrhage in the lateral ventricle and corpus callosum on the percussion side. The severity of pathologic changes increased with the magnitude of percussion. The results indicate that the new device could inflict reproducible graded lateral fluid percussion brain injury on rats and the model can be used for the studies of neuropathologic and molecular mechanism of brain injury. PMID- 11387941 TI - [The activities of proteolytic enzymes in dental plaque of patients in different age-sex groups]. AB - This study was intended to explore the relationship between the proteolytic enzymes in dental plaque and the patient's age and sex. Fluorometry and spectrometry were used to separately determine the activities of leucine amino peptidase, dipeptidyl peptidase IV and trypsin-like proteinase in children(10 boys and 10 girls), adults(n = 20) and elderly people(n = 10). The results showed that sex almost had no influence on the activities of the three enzymes in dental plaque in the children(P > 0.05), and with the increase of age, the activities of the three enzymes apparently increased (P < 0.01). These findings imply that the proteolytic enzymes may only play important roles in root lesions. PMID- 11387942 TI - [Effect of human plasma HDL on the HDL receptors of plasma membranes of cholesterol-fed rabbit]. AB - Atherosclerosis(As) rabbit model was developed by high-cholesterol feeding for 12 weeks. The rabbits were injected intravenously with human plasma HDL preparation per week, and then the effects of HDL on the lipids contents of serum, liver and bile nd the activity of HDL receptors on liver plasma membranes of cholesterol fed rabbit were investigated. The results showed that HDL preparation had no effect on decreasing the lipids contents of serum, but it could low down the lipid depositions in liver, and promote the excretion of lipids from bile. The value of Bmax of HDL receptor showed decreasing trendy and the value of Kd showed increasing trendy in cholesterol-fed rabbits. In HDL-treated rabbits, the value of Bmax increased significantly as compared with that of normal group(P < 0.05), but the value of Kd showed no difference. The results suggested that human plasma HDL could enhance the activity of HDL receptors on the liver plasma membranes of cholesterol-fed rabbit. PMID- 11387943 TI - [Apolipoportein E polymorphism, serum lipids and apolipoproteins of 362 Han national subjects in Chengdu area]. AB - This investigation was conducted to observe the frequency distribution of apoE phenotypes and alleles and to explore the relationship between apoE polymorphism and plasma lipids or apolipoproteins in Chinese population. ApoE phenotypes were assayed by isoelectric focusing and immunoblotting with serum. Serum lipids and apoA I, B100, C II, C III, E were determined in a random subset of 362 subjects including 268 males and 94 females with a mean age of 43.7 +/- 12.3 yrs from a population of Han Nationality in Chengdu area. The results showed that the frequencies of apoE phenotypes and alleles were: E3/3 72.93%, E2/3 12.98%, E3/4 11.33%, E2/4 1.38%, E4/4 1.38%, E2/2 0.00%; epsilon 3 0.8508, epsilon 2 0.0718, epsilon 4 0.0774. The results also showed that the apo E2(E2/3 + E2/2) group had lower levels of serum TC and apoB100 (P < 0.05) and a higher level of serum apoE (P < 0.001) when compared with the apoE3(E3/3) or apoE4(E3/4 + E4/4) group. No significant difference was observed in TG, apoA I, apoC II, and apoC III levels among the apoE2, E3 and E4 groups (P > 0.05). PMID- 11387944 TI - [Construction of nested set of unidirectional deletion of recombinant plasmid DNA for sequencing]. AB - In order to rapidly sequence a batch of large DNA fragments, we developed a method for the construction of nested set of unidirectional deletions. A nested set of unidirectional deletions of the large DNA fragment of a c-type retrovirus gene was successfully constructed by the adoption of this method. The recombinant plasmid DNA was excised by BamHI and SphI at on end of the target DNA to create 5' and 3' overhanging ends. The DNA was digested with Exo III from 5' overhanging end to generate a set of unidirectional deletion. With the use of this method a set of deleted plasmid DNA and the deleted target DNA excised from the deleted constructs were observed on agrose gel electrophoresis. We sequenced all of 24 deleted DNA fragments (forward and reverse orientation). All of them contained overlapping sequences at their ends. These sequences were readily aligned. The results demonstrate this is an efficient method for sequencing large DNA fragment. The influential factors in this method were discussed in this report. PMID- 11387945 TI - [Intrapulmonary expression of tumor necrosis factor alpha mRNA and its significance in rats with acute pancreatitis]. AB - It is thought that tumor necrosis factor (TNF) plays an important role in pathogenesis of acute lung injury or ARDS. So we want to get insight into the relationship between intrapulmonary expression of TNF-alpha gene and lung injury during acute necrotizing pancreatitis (ANP). In our study, acute edematous pancreatitis (AEP) and ANP were induced in rats by caerulein and sodium taurocholate respectively. After acute pancreatitis was induced, serum TNF-alpha was assessed by ELISA assay while endotoxin was assessed by limulus lysate test. Intrapulmonary expression of TNF-alpha mRNA was detected by semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) technique. Moreover, content of pulmonary lesion was investigated and graded with microscope. It was found that TNF-alpha concentration in blood elevated markedly after acute pancreatitis was induced, especially in ANP group. Results of RT-PCR revealed that no TNF-alpha mRNA could be detected in lung tissue from those rats undergoing sham-operation, but marked expression appeared 1 hour after AEP or ANP was induced. Upregulation of expression of TNF-alpha gene in the early 3 hours was similar in the two groups with pancreatitis, and since then expression of TNF alpha mRNA in ANP group was stronger than that in AEP group. Serum endotoxin increased significantly 6 hours after ANP was induced, with a higher level at 12 hour. However, there was no marked change of endotoxin level in AEP group and control group. It is noted that intrapulmonary expression of TNF-alpha gene in ANP group reached its peak as soon as serum endotoxin increased markedly. Lung damage in ANP group was more serious than that in AEP group significantly. Score of lung injury correlated well with TNF-alpha concentration in blood and expression of its gene in lung tissue in either AEP group or ANP group, as well as with serum endotoxin in ANP group. So overwhelm expression of some harmful cytokines like TNF-alpha in lung tissue may be the main cause of lung injury during acute necrotizing pancreatitis, and stimulation of endotoxiemia can at least partly explain the upregulation of expression of TNF-alpha gene. PMID- 11387946 TI - [Research on stochastic factors of heart rhythm variability in nonlinear dynamic analysis]. AB - In this study, the influence of external noise on heart rhythm variability (HRV) in nonlinear dynamic analysis and the internal stochastic factor of HRV were discussed in connection with their physiological significance. The sampling rate, stationary and non-uniformity time interval were analyzed. An interpolation algorithm for heart period was introduced. The experiment demonstrated that the influence was highly reduced by the mentioned procedure. The internal stochastic factors of HRV were investigated by several nonlinear dynamic methods such as surrogate data, mutual information and nonlinear forecasting. The results suggested the colligation of deterministic chaos and stochastic process should be a favorite to interpret HRV. PMID- 11387947 TI - [Expression of focal adhesion kinase in lymphatic endothelial cells of metastasis]. AB - This study was directed at the expression of focal adhesion kinase (pp125FAK) which is located together with vinculin and talin in the spot of cellular adhesion. The authors' intention was to collect reliable data on this important kinase in the signal pathways so as to provide in-depth materials for exploring the mechanism of tumor metastasis to lymphatic vessels. The immunohistochemical method was used to study the expression of FAK in lymphatic endothelial cells of metastaltci adenocarcinoma of rectum and its adjacent lymph-nodes. The result demonstrated that lymphatic endothelial cells expressed pp125FAK when lymphatic vessels were invaded by cancer cells. Reaction production was located in the cytoplasm. This study provides that there was a strong correlation between the expression of FAK in lymphatic endothelial cells and the metastasis of cancer to lymphatic vessels. PMID- 11387949 TI - [Relationship between expression of P16 protein and prognosis in carcinoma of nasopharynx]. AB - We observed the relationship between p16 gene and the occurrence, development and prognosis of carcinoma of nasopharynx by means of LSAB immunohistochemistry method and investigated the expression of protein P16 in 73 cases of carcinoma of nasopharynx and 10 cases of epithelia of chronic inflammation of nasopharynx. The results showed that P16 positive rate was 100% for the epithelia of chronic inflammation of nasopharynx. It was significantly higher than the P16 positive rate for the carcinoma of nasopharynx (38.4%, P < 0.01). There was significant difference of P16 positive expression in differentiation of nasopharyngeal carcinoma (poor differentiation versus undifferentiation), clinical staging (I-II versus IV) and grading of tumor (T1-T2 versus T3, T4) (P < 0.01). There was no correlation between P16 expression and metastatic lymph node (P > 0.05). The 3 year survival rates were 88.9% and 72.9% in P16 expression (+) and (-) patients respectively (P < 0.05). These findings suggested that P16 might play an important role in the process of carcinogenesis and development of nasopharyngeal carcinoma, and P16 might be used as a valuable marker for assessment of the biological behavior and prognosis of nasopharyngeal carcinoma. PMID- 11387948 TI - [nm23-H1 protein expression and its clinical significance in human laryngeal carcinoma]. AB - The Objective of the study on the expression of nm23-H1 protein in human laryngeal carcinoma, was to explore the role of nm23 gene in lymph node metastasis and invasion. The expression of nm23-H1 protein was immunohistochemically examined in 86 resected human laryngeal carcinomas. Anti nm23-H1 monoclonal antibody was used. The results showed that 66.3%(57/86) of the cases were positive for nm23-H1 protein expression in the primary lesions. The positive staining rate (90.5%, 38/42) of the patients without regional lymph node metastasis, was significantly higher than that of patients with nodal involvement (43.2%, 19/44). Inverse relationship were found between the nm23-H1 protein expression in the primary lesions and the depth of tumor invasion or the advance of clinical stage. Expression of nm23-H1 protein was positively associated with the ascending histological grade of the lesion, and the relationship was significant. The above data suggest that nm23 gene plays an important role in the control of lymph node metastasis and invasion in laryngeal carcinomas. PMID- 11387950 TI - [Experimental study of ta chengchi tang decoction for relieving lung injury during acute necrotizing pancreatitis]. AB - The objectives of the study were to investigate the changes of leukocyte adhesiveness and tumor necrosis factor (TNF) in the early stage of acute necrotizing pancreatitis (ANP), to go further into the relation of those changes to lung injury of ANP, and to evaluate the prohibitive effect of ta chengchi tang Decoction on leukocyte adhesion and TNF secretion. 14 canines were randomly divided into 3 groups. The treatment group (n = 5): experimental ANP model treated with ta chengchi tang Decoction; the saline control group (n = 5): experimental ANP model treated with normal saline; and the sham operative group (n = 4). The adhesion of leukocytes to vascular endothelial cells and TNF activities in serum and in bronchi-alveolar lavage fluid (BALF) were observed and measured, and the degrees of lung injury were assessed. The results showed that the adhesiveness of leukocytes was markedly increased and the activities of TNF were elevated in peripheral blood and BALF during ANP; that the leukocyte adhesiveness in the treatment group was weaker than that in the saline group (P < 0.05) and the serum and BALF TNF activities were also significantly lower than those in the saline group (P < 0.01); and that the degrees of lung injury in the treatment group were significantly milder than those in the saline group. These indicate that the increased leukocyte adhesiveness and hypersecretion of TNF take part in the pathogenesis of lung injury in ANP, and ta chengchi tang Decoction is demonstrated an efficacious medicine for alleviating the degree of lung injury mediated by both the leukocyte adhesion and TNF in ANP. PMID- 11387951 TI - [Apoptosis and necrosis of neuron after focal ischemia in brain of rats]. AB - This article reports the apoptosis and necrosis of neurons following ischemia reperfusion in the brain of rats. The focal ischemia model was established by occluding the middle cerebral artery for 2 h and reperfusing for 0.5-48 h. The neuronal changes were investigated by HE staining, TUNEL and agarose gel electrophoresis. The foci of ischemia were in the preoptic area, striatum and cortex. At 0.5 h of reperfusion, there were quite a few apoptotic neurons in the preoptic ischemic focus, but only a few scattered apoptotic cells were seen in the striatum and cortex. During 3-6 h of reperfusion, the number of apoptotic cells increased and the necrotic cells appeared. The number of apoptotic cells reached a peak at 12-24 h and the in morphology varied, and they were mainly located in the inner boundary of infarct. At 48 h, the apoptotic cells decreased; and the necrotic cells increased. Agarose gel-electrophoresis showed the dense smear and ladder pattern of DNA fragments, the apoptosis and necrosis of neurons coexisted, and apoptosis contributed to the enlargement of the ischemic infarct. PMID- 11387952 TI - [DNase I sensitivity of nucleus and chromatin in six different tissues of old rates]. AB - In order to investigate the mechanism of aging, we studied the DNase I sensitivity of nucleus and chromatin in six different tissue (heart, liver, spleen, lung, kidney and brain of old and weanling wistar rats. The results showed that the DNase I digestive sensitivity levels of nucleus and chromatin in the tissue of the old rats were lower than those of the weanling rats respectively that in the old rats, the DNase I digestive sensitively in the tissue varied, the lowest sensitivity was in brain (P < 0.01); and that in the weanling rats, lowest DNase I digestive sensitivity of nucleus and chromatin was also in the brain, but the difference was not significant. PMID- 11387953 TI - [Significance of maximal midexpiratory time and F-V curve configuration in testing small airway function for smokers]. AB - To evaluate the clinical significance of maximal midexpiratory time (MET) in testing small airway functions for smokers, the routine ventilatory functions, MET and F-V curve configurations were studied in 116 passive and light, heavy active smokers. Routine ventilatory functions and F-V curve configuration were determined with LR80 flow volume recorder. The MET was measured with FJD80 spirometer. The results showed that the distribution of convex type rose with the degree of smoking. In heavy smokers, the MET elongated to 127% of the predicted value and in the convex type F-V curve, it elongated to 120% of the predicted value; both were consistent with the decrease of corresponding V50, V25. This implied when MET elongated to 120%-130% of the predicted value in smokers, there had been slight disturbances in small airway functions. It is therefore suggested that MET and convex type F-V curve both are sensitive indices for testing small airway functions, thus it should be of clinical significance and be appropriate for general application. PMID- 11387954 TI - [Pharmacokinetics of pramiracetam in animals]. AB - Pharmacokinetic rules of pramiracetam were studied here. After giving pramiracetam orally to dogs, we drew their blood at various times. The drug concentrations in blood plasma were detected by HPLC. 3p87 program was used to calculate the pharmacokinetic parameters. The time-concentration curve corresponded to one apartment model. T1/2 was about 2.3-3.9 hours in various doses. After pramiracetam was given to rats per os, high concentrations of pramiracetam were detected in the rats' tissues. The kidney had the highest concentration of pramiracetam; the liver had the next highest concentration, and then the intestine, lung, muscle, heart, gonad, spleen and sebum had the high concentration in order. The drug was also detected in the brain. 0.7% of the given dose was excreted in unchanged form in bile in 24 hours. 28.26% and 6.35% were excreted in urine and feces respectively in 72 hours. The plasma protein combining rate detected by the method of balance dialysis was 20.1-22.2%. PMID- 11387955 TI - [A quantitative histological investigation of salivary glands of human and experimental animals]. AB - The pairs of three major salivary glands of eight kinds of full grown experimental animals were sectioned and examined by morphometric techniques. The televised image-analysis system was used to determine the acinar area and proportional volumes of component tissue. The results were compared with those of the same glands from human. There were no differences between the three acinar areas of human and monkey. The acinar area of human parotid was the smallest except that of rabbit. The acinar area of human submandibular gland was larger than that of rodents but smaller than that of other animals. There were no differences between the acinar area of sublingual glands of human and animal except those of rabbit and goat. The acinar area of mucous acini was larger than that of serous and seromucous acini. The mean proportional volume of acini of human parotid was less than that of other animals. The duct systems of rodents submandibular were far more than those of other animals. In sublingual glands, there were no differences in mean proportional volume of component tissue between human and animals except there were more acini and less fiber in rodents' subingual glands. PMID- 11387956 TI - [The effect of vasectomy on spermatogenesis and proliferation of rat spermatogenic cell]. AB - In order to explore the effect of vasectomy on spermatogenesis and proliferation of rat spermatogenic cell, we detected the DNA content of rat spermatogenic cells by flow cytometry at 4 weeks and 8 weeks after vasectomy. The results showed that the percentages of 1C cell group (spermatid and sperm) in the two experimental groups were apparently lower than that of the two control groups respectively (P < 0.05), but the percentage of 2C cell group (spermatogonium and spermotocyte) in the two experiment groups were much higher than those of the two control groups respectively (P < 0.05); The proliferation index (PI) of experimental groups was significantly lower than that of control groups (P < 0.05) and most of the cells in the experimental groups were on G0/G phase. The percentage of S-phase cell in the 8 weeks experimental group was obviously lower than that of the 4 weeks experimental group. These findings suggest that vasectomy in rat may affect many stages in spermatogenesis and delay the proliferation of spermatogenic cell. PMID- 11387957 TI - [A study on the T-cell subsets and glucocorticoid receptor in children with anaphylactoid purpura]. AB - To study the changes of T-cell subsets and glucocorticoid receptor (GCR) in children with anaphylactoid purpura. T-cell subsets and GCR were measured respectively by cytotoxicity assay and GCR radioligand-binding assay in peripheral blood lymphocytes from 35 children with anaphylactoid purpura and 35 normal children. The result showed that the average percentages of CD3+, CD4+ cell subtypes and the CD4+/CD3+ ratio were 46.03% +/- 9.40%, 31.06% +/- 6.80% and 1.23 +/- 0.33 respectively, and the GCR numbered 3060 +/- 2153 binding sites per cell in the study group, on the other hand, the average percentages of CD3+, CD4+ cell subtypes and the CD4+/CD3+ ratio were 53.11% +/- 5.40%, 35.01% +/- 4.41% and 1.52 +/- 0.26 respectively, and the GCR numbered 5210 +/- 1639 binding sites per cell in the control group. These indicate that and that patients with anaphylactoid purpura may have abnormal immunomodulations and decreased GCR numbers per cell, which may have effects on the immune dysfunctions in the pathogenesis of anaphylactoid purpura. PMID- 11387958 TI - [Measurement and clinical use of recoil pressure of the respiratory system]. AB - Using the principle that the difference of MIP (maximal inspiratory mouth pressure) between RV (residual volume) and FRC (functional residual capacity) equals Prs (recoil pressure of the respiratory system) at RV, we performed the measurement of Prs at RV in 20 normal subjects, 90 patients with COPD (chronic obstructive pulmonary disease) and 15 patients with ILF (interstitial lung fibrosis). We also performed the correlation analysis of Prs at RV and RV/TLC (total lung capacity) %. The results indicated the Prs values at RV (-2.8 +/- 0.5 kPa) in normal subjects measured in this manner was in accord with the values reported by other authors abroad. The Prs in patients with emphysema decreased and it was highly and negatively correlated with RV/TLC% (r = -0.872), but the Prs in patients with ILF was increased. The results suggest that this method for measurement of Prs is simple, reliable, and clinically useful. PMID- 11387959 TI - [Experimental study of the effect of dexamethasone on pulmonary diffuse hemorrhage and energy metabolism of mitochondria in the liver of guinea pig infected with leptospirosis]. AB - This experiment was designed to study the curative effect of dexamethasone on leptospirosis with pulmonary diffuse hemorrhage (PDH) in guinea pig model. The guinea pigs were randomly divided into four groups: the infected group, all animals developed PDH(n = 10); the infected plus treated group(n = 10); the treated control group(n = 10); and the normal control group(n = 10). The gross and microscopic observations in lung, liver and kidney were made. Also, ultrastructural changes of mitochondria in liver were examined, and the lactic acid level and mitochondrial ATPase activity in liver were determined. The results showed that there was significant difference in mortality, in gross and microscopic changes in lung, liver and kidney, and in the changes of mitochondrial ultrastructures, ATPase activity and lactic acid level in liver between the infected group and the infected plus treated group (P < 0.01). These findings suggest that the pharmacologic dose of dexamethasone injected intravenously in the early stage of PDH may have some beneficial effect on the emergency treatment of leptospirosis with PDH. The mechanism appears to be related to its effect on the inhibition of glycolysis and increasing oxidative phosphorylation in mitochondria. PMID- 11387960 TI - [Effects of protocol HX-97 on mobilization, collection and hematopoietic reconstitution of peripheral blood stem cells transplantation]. AB - In order to enhance the effects of peripheral blood stem cell mobilization, collection and hematopoietic reconstitution, we observed and evaluated the effects of Protocol HX-97 on 22 patients who received allogeneic or autologous peripheral blood stem cell transplants from April 1997 to June 1999. rhG-CSF was used for mobilization at a dose of 300 micrograms/day for 6 days; the sixth dose was given an hour and a half before leukapheresis. High dose chemotherapy and rhG CSF were given for autologous peripheral blood stem cell mobilization; the chemotherapy should be intensive enough to reduce the patients' peripheral WBC to less than 1.0 x 10(9)/L, and the beginning of using rhG-CSF should be just at the time of WBC's rising from the nadir. rhGM-CSF and rhG-CSF were given sequentially for hematopoietic reconstitution after transplantation. The results showed that leukaphereses were successfully performed for peripheral blood stem cell collection. Sixteen cases needed apheresis only once, and 6 cases needed it twice. The harvests were 2.5-10.7 x 10(8)/kg MNC, 2.5-20.0 x 10(6)/kg CD34+ cells (including 1.8-7.5 x 10(6)/kg CD34+CD33-, 0.7-12.5 x 10(6)/kg CD34+CD33+), and 3.5-6.3 x 10(5)/kg CFU-GM. Hematopoietic function recovered to > 0.5 x 10(9)/L of neutrophil count in allo-PBSCT at 14-20 days and in auto-PBSCT at 12-20 days, and to > 20 x 10(9)/L of platelet count in allo-PBSCT and in auto-PBSCT at 16-34 days and 16-28 days, respectively. At day +30 post-transplantation, chromosome analysis and DNA finger print assessment of bone marrow cells indicated that the patients' hematopoietic function had been reconstituted. This study suggests that Protocol HX-97 is an effective approach to mobilization of peripheral blood stem cell transplantation, and it is relatively cost-effective. PMID- 11387961 TI - [GB virus-C infection in Chinese patients with hepatocellular carcinoma]. AB - Hepatitis B and C viruses (HBV, HCV) are closely related with hepatocellular carcinoma(HCC). The association between the new discovered GB-virus C (GBV-C) and HCC has not yet been known. In this study, 124 HCC patients were detected for the prevalence of GBV-C RNA by the one-step nested reverse transcription polymerase chain reaction (RT-PCR) and followed by hybridization using GBV-C probes located at 3'-untranslated region (3'-UTR) from its reported genomes. The results showed that 33 of 124 (26.6%) HCC cases were GBV-C RNA positive, including 12 cases positive HBsAg and anti-HCV, and 3 for cases negative HBsAg and anti-HCV. The clinical background of the patients with HBsAg and/or anti-HCV who were also positive for GBV-C RNA did not differ from the background of those who were negative for GBV-C RNA except the ratio of blood transfusion history. In conclusion, GBV-C has a high prevalence in Chinese HCC patients. Even though no sufficient data supports the causality of GBV-C on hepatocarcinogenesis, further researches aimed at whether GBV-C infection aggravates the incidence of HCC are warranted. PMID- 11387962 TI - [Evaluation of effectiveness of hepatitis B plasma derived vaccine and a survey of hepatitis C virus infection in preschool children in Chengdu, Sichuan]. AB - This study was undertaken to evaluate the effectiveness of hepatitis B plasma derived vaccine and investigate the character of hepatitis C virus infection in preschool children. The anti-HBs, HBsAg and anti-HCV of 985 plasma samples were examined by ELISA. The results showed that the positive rate of HBsAg in the children (aged < 7 yr) in the communities was 0.89% and the effectiveness of the vaccine was 91.38%. The positive rate of anti-HBs decreased with the increase in age; the positive rate of HBsAg did not increase with age. The positive rate of HBsAg was as high as 4.69% in the children from hospitals, so supervision and enhanced vaccination should be administered properly in this group. The positiverate of anti-HCV was 1.18% in the communities, but 3.17% in hospitals. It is worthy to investigate how children get infected with HCV. PMID- 11387963 TI - [Clinical value of 99mTc-MIBI imaging, 131I whole body scan and HTG determination for the follow-up of patients with differentiated thyroid carcinoma after 131I therapy]. AB - The purpose of this study was to make a comparative evaluation of 99mTc-MIBI imaging, 131I whole body scan and HTG determination in use for the follow-up of patients with differentiated thyroid carcinoma (DTC) after 131I therapy. Fifty three patients with DTC had undergone surgical treatment and 131I ablation. Clinical examinations showed that 22 of them had metastases or recurrences. In the follow-up of all the patients, the HTG determination, the 99mTc-MIBI imaging and the 131I-WBS were performed in sequence after the discontinuance of the administration of thyroxin for 4-6 weeks. The results showed that the sensitivity, specificity and accuracy of 99mTc-MIBI imaging were 86.36%, 87.10% and 86.79% respectively; those for HTG were 68.18%, 100% and 86.79% and those for 131I-WBS were 63.64%, 100% and 84.91% respectively. The total sensitivity of the three examinations was 100%; the total specificity and total accuracy were 87.10% and 92.45% respectively. The data of this study suggest that 99mTc-MIBI imaging is a highly sensitive method and the combined use of the three examinations can improve the sensitivity and accuracy of diagnosis for patients with DTC. PMID- 11387964 TI - [A study of color Doppler ultrasound on arteries of the extremities in patients with type 2 diabetes]. AB - To investigate the changes of the arteries in extremities in patients with type 2 diabetes(DM2), fifty patients with DM2(35 women and 15 men, mean aged 57.54 +/- 14.19 years old, time of DM2 diagnosed one week-26 years, without the histories of hypertension and smoking) were studied. Radial, finger, anterior tibial and dorsum pedis arteries of all subjects were examined using color Doppler Ultrasonography, 30 of 50 patients had the symptoms of extremities (e.g., numbness, coldness and pain). The Doppler examination revealed: 1. In the patient group, arterial wall became thick, rough and rigid. The atherosclerotic plaques were found inside vascular cavities in 14/50 in patient group with one patient being asymptomatic of extremities. 2. The vascular cavities in patients remarkably narrowed compared with the control group (P < 0.05). The vascular lesion worsened along with the development of the symptoms. 3. Doppler spectra in the majority of patients displayed single-peak, whereas it displayed three peaks in normal subjects with significant differences (P < 0.05). 4. The peak flow velocities in the patients increased, and this increase was related to the development of the symptoms, but no significant differences were found. Color Doppler Ultrasound examination is valuable in the evaluation of the arterial lesion of the extremities, especially at the early stage of artery diseases in patients with DM2. PMID- 11387965 TI - [Effects of lovastatin on plasma lipid, plasma glucose and insulin metabolism of patients with type II B hyperlipemia]. AB - This study was directed at the effects of lovastatin on plasma glucose and insulin metabolism of patients with Type II B hyperlipemia. Thirty patients with Type II B hyperlipemia were observed. At the beginning, We detected the patients' TC, TG, HDL-C, VLDL-C with enzyme assay, calculated their LDL-C with formula, determined their plasmaglucose(PG) with oxidase assay and plasma insulin(PI) with immunoradioassay, and then we performed OGTT and insulin release test, these indices were detected again after the administration of 8 weeks' lovastatin therapy (20 mg per day). The results showed that, aftert the therapy, TC and LDL C decreased significantly(P < 0.05), TG, HDL-C and VLDL-C had no significant change(P > 0.05). Fasting plasma glucose(FPG) and fasting plasma insulin(FPI) increased (the former, P < 0.05). Two hours after administration of glucose, the level of plasma glucose(2hPG) and plasma insulin(2hPI)increased(the latter, P < 0.05). 1/(FPG x FPI) had no significant change, but 1/(2hPG x 2hPI) decreased (P < 0.05). These data indicate that long-period use of lavastatin may cause insulin resistance. So we advise patients with Type II B hyperlipemia treated by statin to limit their glucose intake in case of hyperinsulinemia and insulin resistance. PMID- 11387966 TI - [The diagnostic value of hysteroscopy on postmenopausal bleeding]. AB - The objectives of this study were to investigate the causes of postmenopausal bleeding and evaluate the diagnostic value of hysterscopy. Using the diagnostic technique of hysteroscopy and directed biopsies, the authors analysed the causes of postmenopausal bleeding of 234 patients undergoing hysteroscopy. The results showed that atrophic endometrium was the most common cause of postmenopausal bleeding and that endometrial hyperplasia, endometrial carcinoma/precancerous lesion, and submucous leiomyoma/endometrial polyp were also common in the patients with postmenopausal bleeding. The agreement rate between hysteroscopy and directed biopsies was 74.61%. In 234 cases of postmenopausal bleeding, the benign and malignant pathological changes were found in 76 and 27 cases respectively. In terms of age and menopause time, there were significant differences (chi 2 = 3.36, P < 0.01; chi 2 = 3.92, P < 0.01 respectively)between the patients with postmenopausal bleeding in the benign changes group and those in the malignant changes group. So the combined use of hysteroscopical technique and directed biopsies is of great significance in finding out the causes of postmenopausal bleeding, making a definite diagnosis of endometrial carcinoma and precancerous lesion, and further preventing the occurrence of endometrial carcinoma. The examination of hysteroscopy is an effective and valuable method for diagnosing postmenopausal bleeding. PMID- 11387967 TI - [Changes and significance of natural killer cell, IL-2, IL-6 and TNF alpha of heroin addicts after detoxification]. AB - In order to find out the activity of NKC and production of IL-2 in the peripheral blood mononuclear cells and the level of IL-6 and TNF alpha in the peripheral blood serum, we investigated 51 herion addicts who were in the course of detoxification. MTT colorimetric assay and ELISA assay methods were adopted. The results showed that, in the herion addicts, NK activity was 30.11 +/- 5.2%, IL-2 activity was 8.06 +/- 1.66 IU/ml, IL-6 level was 61.17 +/- 12.07 pg/ml, and TNF alpha level was 91.83 +/- 19.19 pg/ml, whereas in the healthy controls, the results were 44.89 +/- 4.75%, 15.91 +/- 3.83 IU/ml, 22.18 +/- 9.31 pg/ml and 30.55 +/- 11.94 pg/ml respectively. When the herion addicts were subjected to detoxication, their NKC and IL-2 activities, IL-6 and TNF alpha levels gradually restored to normal, and were correlated with the time of their detoxification (NKC: r = 0.626, P < 0.001; IL-2: r = 0.684, P < 0.001; IL-6: r = -0.791, P < 0.001; TNF alpha: r = -0.703, P < 0.001). PMID- 11387968 TI - [Determination of 6-methoxy-2-naphthylacetic acid, a major metabolite of nabumetone, in human plasma by HPLC]. AB - This paper reports a sensitive and rapid method for determining 6-methoxy-2 naphthylacetic acid (MNA), a major metabolite of nabumetone in human plasma using naproxen as the internal standard. High performance liquid chromatograph model 680 (Waters, USA) with a variable wavelength UV detector and reversed-phase YWG C18 column (10 microns, 250 x 4.6 mm) was used. After the addition of acetate buffer(pH3.0), the plasma sample was extracted with methylene chloride. The mobile phase of methanol-pH3.0, 0.02 mol/L acetate buffer(74:26) was pumped at 1.0 ml/min through the column. The detector at 0.01AUFS was set at 270 nm. The retention times for MNA and naproxen were 3.98 min and 4.73 min respectively. Standard curve was linear in the concentration range of 0.5 to 64 mg/L. The detection limit in serum was 0.02 mg/L. Extraction recovery was 88%-94%; method recovery 96%-102%; withinday RSD less than 3.5%; inter-day RSD less than 5%. PMID- 11387969 TI - [One simple and efficient method for purification of IgG McAb from mice ascites: caprylic acid/ammonium sulfate precipitation]. AB - Several ascitic IgG McAbs were purificated by caprylic acid/ammonium sulfate precipitation (CA-AS), ammonium sulfate precipitation (AS) and SPA affinity chromatography. Purity of IgG purificated by CA-AS ranged from 87% to 91%, higher than that obtained by CA (65%-70%), lower than that obtained by SPA affinity chromatography (100%). The yield of IgG purificated by CA-AS ranged from 4.8 g/L to 5.5 g/L, lower than that obtained by CA (7.0-7.9 g/L), more higher than that obtained by SPA affinity chromatography (0.6 g/L). The recovery of IgG purificatied by CA-AS ranged from 39.0% to 44.4%, lower than that obtained by CA (56.0%-65.0%), more higher than that obtained by SPA affinity chromatography (7.4%). The ELISA titer of 4E2 or H6McAb was not impaired by purification of CA AS. The above results suggested that CA-AS is one simple, efficient, rapid and low-cost method for purification of IgG McAb from ascitic fluid. PMID- 11387970 TI - [Simultaneous determination of norepinephrine and epinephrime in plasma by using high performance liquid chromatograph]. AB - A sensitive and rapid method was reported for the determination of nerepinephrine (NE) and epinephrine (E) in plasma by using high-performance liquid chromatography with an electrochemical detector (HPLC-EC) and YWG-C18 column(5u). 3,4-Dihydroxybenzylamine(DHBA) was used as an internal standard. The mobile phase of 0.05 mol/L actate buffer(pH3.0)-methanol-0.05 mol/L EDTA-Na2(78:20:2) with of 200 mg/L SDS solution was pumped through the column with a flow rate of 1.0 ml/min. The lengths of retention time for NE, E and DHBA(IS) was 7.3 min, 8.3 min and 10.4 min, respectively. The standard curve was linear within the concentration range of 31.25 to 4000 ng/L for NE and E (NE: r = 0.9999 E: r = 0.9996). The within day RSD (%) were less than 1.87% for NE and less than 2.07% for E, inter-day RSD(%) were less than 8.92% for NE and less than 9.65% for E respectively. The average extraction recovery(%) for NE and E were 93.5% +/- 5.50% and 91.3% +/- 1.77% respectively. This method has been applied to the diagnosis of pheochromocytoma and clinical research. PMID- 11387971 TI - Opening address: the challenge to feed the World's poor. PMID- 11387973 TI - Rice biotechnology: improving yield, stress tolerance and grain quality. Proceedings of a symposium. Laguna, Philippines, 27-29 March 2000. PMID- 11387972 TI - Introduction of genes encoding C4 photosynthesis enzymes into rice plants: physiological consequences. AB - Transgenic rice plants expressing the maize phosphoeno/pyruvate carboxylase (PEPC) and pyruvate, orthophosphate dikinase (PPDK) exhibit a higher photosynthetic capacity (up to 35%) than untransformed plants. The increased photosynthetic capacity in these plants is mainly associated with an enhanced stomatal conductance and a higher internal CO2 concentration. Plants simultaneously expressing high levels of both enzymes also have a higher photosynthetic capacity. The results suggest that both PEPC and PPDK play a key role in organic acid metabolism in the guard cells to regulate stomatal opening. Under photoinhibitory and photooxidative conditions, PEPC transgenic rice plants are capable of maintaining a higher photosynthetic rate, a higher photosynthetic quantum yield by PSII and a higher capacity to dissipate excess energy photochemically and non-photochemically than untransformed plants. Preliminary data from field trials show that relative to untransformed plants, the grain yield is about 10-20% higher in selected PEPC and 30-35% higher in PPDK transgenic rice plants, due to increased tiller number. Taken together, these results suggest that introduction of C4 photosynthesis enzymes into rice has a good potential to enhance its tolerance to stress, photosynthetic capacity and yield. PMID- 11387974 TI - Increasing rice photosynthesis by manipulation of the acclimation and adaptation to light. AB - There are three important considerations in assessing the interaction of crop plants with light: (a) how does the plant respond to the light environment both in the short-term (regulation) and in the long-term (acclimation), (b) under what conditions are these responses inadequate, leading to photoinhibition, and (c) are the responses optimally adapted for maximum agricultural yield? Despite a wealth of knowledge about these processes in model plant species, it is impossible to predict how significant they are in influencing the yield of rice. Therefore, in collaboration with IRRI, we have undertaken a study of photoinhibition and photoacclimation of rice under field conditions. The results of this study are presented, along with an assessment of the implications for improvement of rice yield. PMID- 11387975 TI - A framework for sequencing the rice genome. AB - Rice is an important food crop and a model plant for other cereal genomes. The Clemson University Genomics Institute framework project, begun two years ago in anticipation of the now ongoing international effort to sequence the rice genome, is nearing completion. Two bacterial artificial chromosome (BAC) libraries have been constructed from the Oryza sativa cultivar Nipponbare. Over 100,000 BAC end sequences have been generated from these libraries and, at a current total of 28 Mbp, represent 6.5% of the total rice genome sequence. This sequence information has allowed us to draw first conclusions about unique and redundant rice genomic sequences. In addition, more than 60,000 clones (19 genome equivalents) have been successfully fingerprinted and assembled into contigs using FPC software. Many of these contigs have been anchored to the rice chromosomes using a variety of techniques. Hybridization experiments have shown these contigs to be very robust. Contig assembly and hybridization experiments have revealed some surprising insights into the organization of the rice genome, which will have significant repercussions for the sequencing effort. Integration of BAC end sequence data with anchored contig information has provided unexpected revelations on sequence organization at the chromosomal level. PMID- 11387976 TI - Increasing rice productivity and yield by manipulation of starch synthesis. AB - Plant productivity and yield are dependent on source-sink relationships, i.e. the capacity of source leaves to fix CO2 and the capacity of developing sink tissues and organs to assimilate and convert this fixed carbon into dry matter. Studies from our laboratories as well as others have demonstrated that rice productivity and yield are mainly sink-limited during its development because of limited capacity to utilize the initial photosynthetic product (triose phosphate). This limitation in triose phosphate utilization, evident at both the vegetative and reproductive stages of rice development, may be associated with limited capacity for carbohydrate synthesis in rice leaves (which are poor accumulators of starch) or feedback due to limited sink strength of developing seeds. Strategies in improving triose phosphate utilization by enhancing starch production in leaves and developing seeds by the expression of engineered genes for ADP glucose pyrophosphorylase, a key regulatory enzyme of starch biosynthesis, are discussed. PMID- 11387977 TI - Genetic analysis of plant disease resistance pathways. AB - Plant disease resistance (R) genes are introduced into high yielding crop varieties to improve resistance to agronomically important pathogens. The R gene encoded proteins are recognitionally specific, interacting directly or indirectly with corresponding pathogen avirulence (avr) determinants, and are therefore under strong diversifying selection pressure to evolve new recognition capabilities. Genetic analyses in different plant species have also revealed more broadly recruited resistance signalling genes that provide further targets for manipulation in crop improvement strategies. Understanding the processes that regulate both plant-pathogen recognition and the induction of appropriate defences should provide fresh perspectives in combating plant disease. Many recent studies have utilized the model plant, Arabidopsis thaliana. Here, mutational screens have identified genes that are required for R gene function and for restriction of pathogen growth in compatible plant-pathogen interactions. Genetic analyses of these plant mutants suggest that whilst signalling pathways are conditioned by particular R protein structural types they are also influenced by pathogen lifestyle. Two Arabidopsis defence signalling genes, EDS1 and PAD4, are required for the accumulation of salicylic acid, a phenolic molecule required for systemic immunity. The cloning, molecular and biochemical characterization of these components suggests processes that may be important in their disease resistance signalling roles. PMID- 11387978 TI - Regulation of systemic acquired resistance by NPR1 and its partners. AB - The NPR1 protein of Arabidopsis thaliana has been shown to be an important regulatory component of systemic acquired resistance (SAR). Mutations in the NPR1 gene block the induction of SAR by the signal molecule salicylic acid (SA). NPR1 contains an ankyrin repeats and a BTB domain which are involved in interaction with other protein(s). To further study the function of NPR1 and the regulatory mechanism of SAR, we used both molecular and genetic approaches to identify additional SAR regulatory components. Through a yeast two-hybrid screen we found that NPR1 interacts specifically with bZIP transcription factors. The involvement of bZIP transcription factors in controlling the SA-induced genes had been suggested by a number of promoter studies performed on these genes. It was found that as1 element, which is a binding site for bZIP transcription factors, is essential for SA-induced gene expression. In a genetic screen for suppressors of npr1, we found a mutant, sni1, that restored the responsiveness to SAR induction in npr1. The genetic characteristics of the sni1 mutant and the sequence of SNI1 suggest that the wild-type SNI1 protein is a negative regulator of SAR. We believe that SAR is controlled by both positive regulators and negative regulators. PMID- 11387979 TI - Improving plant drought, salt and freezing tolerance by gene transfer of a single stress-inducible transcription factor. AB - Plant productivity is greatly affected by environmental stresses such as drought, salt loading and freezing. We reported that a cis-acting promoter element, the dehydration response element (DRE), plays an important role in regulating gene expression in response to these stresses in Arabidopsis. The transcription factor DREB1A specifically interacts with the DRE and induces expression of stress tolerance genes. We show here that overexpression of the cDNA encoding DREB1A in transgenic Arabidopsis plants activated the expression of many of theses stress tolerance genes under normal growing conditions and resulted in improved tolerance to drought, salt loading and freezing. However, use of the strong constitutive 35S cauliflower mosaic virus (CaMV) promoter to drive expression of DREB1A also resulted in severe growth retardation under normal growing conditions. In contrast, expression of DREB1A from the stress-inducible rd29A promoter gave rise to minimal effects on plant growth while providing an even greater tolerance to stress conditions than did expression of the gene from the CaMV promoter. As the DRE-related regulatory element is not limited to Arabidopsis the DREB1A cDNA and the rd29A promoter may be useful for improving the stress tolerance of agriculturally important crops by gene transfer. PMID- 11387980 TI - Dissection of defence response pathways in rice. AB - The cloning of major resistance genes has led to a better understanding of the molecular biology of the steps for induction of resistance, yet much remains to be discovered about the downstream genes that collectively confer resistance, i.e. the defence response (DR) genes. We are dissecting the pathways contributing to resistance in rice by identifying a collection of mutants with deletions or other structural rearrangements in DR genes. The collection of rice mutants has been screened for many characters, including increased susceptibility or resistance to Magnaporthe grisea and Xanthomonas oryzae pv. oryzae. A collection of enhanced sequence tags (ESTs) and putative DR genes has been established to facilitate detection of mutants with deletions in DR genes. Arrays of DR genes will be used to create gene expression profiles of interesting mutants. Successful application of the mutant screen will have broad utility in identifying candidate genes involved in disease response and other metabolic pathways. PMID- 11387981 TI - Breeding for nutritional characteristics in cereals. AB - Extensive genetic variation within large species such as the major cereals can be confidently expected for any new trait of interest. This has now been extensively demonstrated for the nutrient content of cereal grains that is of interest under deficient conditions both to human nutritionists and to cereal agronomists. As cereals are eaten in large quantity by practically everyone, they are the ideal vehicles for changing the balance of nutrient intake of the whole human population. Doing so appears to be necessary as the World Health Organization has identified deficient micronutrient intake in well over half of all people globally, notably women and children. Of major concern are iron, zinc, selenium iodine, calcium and vitamin A-related carotenoids. Our results show that for any staple so far studied, the intake of iron, calcium and zinc from cereals can be doubled, and the content/intake of essential carotenoids can be increased by much greater factors. To prove to rigid scientific standards that greater intake results in greater absorption and measurable health benefits is quite difficult, but it is currently being pursued in various ways. This proof of bioavailability is all that impedes implementation in breeding programs. PMID- 11387982 TI - Biosynthesis of beta-carotene (provitamin A) in rice endosperm achieved by genetic engineering. AB - To obtain a functioning provitamin A (beta-carotene) biosynthetic pathway in rice endosperm, we introduced in a single, combined transformation effort the cDNAs coding for (1) phytoene synthase (psy) and (2) lycopene beta-cyclase (beta-lcy; both from Narcissus pseudonarcissus and both under control of the endosperm specific glutelin promoter), with (3) a bacterial phytoene desaturase (crtI, from Erwinia uredovora under constitutive 35S promoter control). This combination covers the requirements for beta-carotene synthesis, and yellow, beta-carotene bearing rice endosperm was obtained in the T0 generation. However, further experiments revealed that the presence of beta-lcy was not necessary, since psy and crtI alone were able to drive beta-carotene synthesis as well as the formation of further downstream xanthophylls. This finding could be explained if these downstream enzymes are either constitutively expressed in rice endosperm or are induced by the transformation, e.g. by products derived therefrom. Based on results in N. pseudonarcissus as a model system, a likely hypothesis can be developed that trans lycopene or a trans lycopene derivative acts as an inductor in a kind of feedback mechanism stimulating endogenous carotenogenic genes. PMID- 11387983 TI - Developing transgenic grains with improved oils, proteins and carbohydrates. AB - DuPont has developed cereals and oilseeds with improved proteins, carbohydrates, and oils for food, feed, and industrial applications. Products which have been or will be introduced include corn and soybeans with increased oil content, improved oil composition, increased amino acid content, altered protein content and functional qualities, altered starch composition, reduced oligosaccharide content, increased sucrose content, and combinations of these traits. These products have been developed using both mutation breeding and molecular biology based transgenic approaches. We have also worked on improving the underlying technologies in order to accelerate product introductions. Gene discovery has been expedited through a genomics program that now has a database of more than two million sequences from a variety of plants, insects and microbes. Plant cell transformation for elite lines of crop species is being addressed through production laboratories with high throughput processes and through technology improvements. High-throughput, rapid and small-scale assays for biochemical parameters are used to identify plants carrying traits of interest. Small-scale functionality analyses, in which grains are broken down into their component parts and assayed for functional properties, indicate which seeds carry a trait of commercial value. Finally, a number of DNA marker systems are being used to accelerate trait introgression timelines. PMID- 11387984 TI - Summing-up: cutting-edge science for rice improvement--breakthroughs and beneficiaries. PMID- 11387985 TI - Rice genomics: current status of genome sequencing. AB - Since its establishment in 1991, the Rice Genome Research Program (RGP) has produced some basic tools for rice genome analysis, including a cDNA catalogue, a genetic linkage map and a yeast artificial chromosome (YAC)-based physical map. For the further development of rice genomics, RGP launched in 1998 an international collaborative project on rice genome sequencing. A P1-derived artificial chromosome (PAC)-based, sequence-ready physical map has been constructed using the PCR markers from cDNA sequences (expressed sequence tag [EST] markers). Selected PAC clones with 100-150 kb inserts from chromosomes 1 and 6 have been subjected to shotgun sequencing. The assembled genomic sequences, after predicting the gene-coding region, have been published both through a public database and through our website. As of January 2000, 1.9 Mb from 13 PAC clones were published. Future prospects for understanding rice genomic information at the nucleotide level are discussed. PMID- 11387986 TI - Rice--the pivotal genome in cereal comparative genetics. AB - Over the past 15 years rice has been the focus of intense co-ordinated research activity to apply the new molecular biology to this key staple. The fact that rice has a small tractable genome and the development of genetic and genomic tools not available in any other cereal have now ensured the promotion of rice as a favoured research target. However the discovery that gene content and gene order--genome colinearity--have been maintained among all the Poaceae family for some 60 million years of evolution has elevated rice yet further to the status of a 'model' organism. Rice tools can be applied in research on the other major cereals, wheat and maize, and many aspects of rice genetics can be transferred to the many minor economic grass species that have not themselves warranted extensive research and breeding. In this paper we describe some of the applications of the discovery of extensive synteny among the grasses. PMID- 11387987 TI - Bioinformatics for rice resources. AB - The distinguishing feature of the 'new biology' is that it is information intensive. Not only does it demand access to and assimilation of vast data sets accumulated by engineered laboratory processes, but it also demands a previously unimaginable level of data integration across data types and sources. There are various information resources available for rice. In addition, there are various information resources that are not focused on rice but that contain rice data. The challenge for rice researchers and breeders is to access this wealth of data meaningfully. This challenge will grow significantly as international efforts aimed at sequencing the entire rice genome come into full swing. Only through concerted efforts in bioinformatics will the power of these public data be brought to bear on the needs of rice researchers and breeders worldwide. These efforts will need to focus on two large but distinct areas: (1) development of an effective bioinformatics infrastructure (hardware systems, software systems, and software engineers and support staff) and (2) computational biology research in visualization and analysis of very large, complex data sets, such as those that will be developed using high-throughput expression technologies, large-scale insertional mutagenesis, and biochemical profiling of various types. In the midst of the large flow of high-throughput data that the international rice genome sequencing efforts will produce, it is also imperative that integration of those data with unique germplasm data held in trust by the CGIAR be a part of the informatics infrastructure. This paper will focus on the state of rice information resources, the needs of the rice community, and some proposed bioinformatics activities to support these needs. PMID- 11387988 TI - Regulation of gene expression by small molecules in rice. AB - A system for the regulation of gene expression by small molecules in transgenic rice was developed. This gene switch system consists of two components: (1) a hybrid chemically activated transcription factor, and (2) a synthetic target promoter. The two elements were transformed into rice suspension cells and transgenic plants were regenerated. A luciferase reporter under control of the gene switch system displayed as high as 10,000-fold inducibility following exposure to the small molecule ligand. The dose-response and induction time course were determined. Regulated luciferase activity in activated plants decreased one day following removal of ligand and could be reactivated multiple times without apparent cosuppression. Analysis of luciferase activity following ligand application to media surrounding the roots suggests that ligand can be absorbed and transported systemically. In contrast, reporter activation was limited to a small area when ligand was applied directly to the leaf surface. The described gene switch system represents an important tool for situations requiring conditional gene expression in a monocot species. PMID- 11387990 TI - [Supplementary ultrasonometry of the fetus in evaluation of gestational age]. AB - The terminology quoted in the leaflet "Ultrasonometry of foetus in evaluation of gestational age" could change the diagnostic and therapeutic procedures in obstetrics from the point of view of only one out of many methods of imaging. The object of the study was to complement the "standards for eleven measurements pertinent to evaluation of pregnancy age and well-being of the foetus", in order to use them in compliance with the laws of auxology with recognize the priority of developmental age over the postmenstrual calendar age. Foetus is not a statistical patient, but an individual one, and it is not clinically important that, e.g. 15% of children are born at the foetal age of 38 or 41 weeks, but whether the examined foetus is born in either of the two weeks. Our complementation should be convincing about the great significance of ultrasonometry not of the foetus (as in the title of the publication in question), but of the foetuses in evaluation of their average gestational age, and about the necessity of reformulation of the published ultrasonomatric standarda, so as to render them clinically useful in regard of the foetuses of various gestational maturation rate. PMID- 11387989 TI - An ecosystem approach to human health. PMID- 11387991 TI - [Evaluation of fetal breathing movements in prognosis of preterm labor]. AB - The relationship between fetal breathing activity and preterm labor among patients with signs of preterm labor has been analyzed. The assessment concerned pregnant women admitted to the hospital because of regular uterine contractions, uterine contractility and bleeding or premature rupture of membranes. It has been found the significant correlation between the absence of fetal breathing movements and preterm labor. PMID- 11387992 TI - [Activity of cathepsin B and cystatins in the placenta during EPH-gestosis]. AB - Activity of cathepsin B using Bz-DL-Arg-pNA contents of SH-group by means of Ellman method, activity of cystatins against papain using casein as a substrate and contents of deoxyribonuclein acids by Burton method were determined in 64 placentas of pregnancies with EPH-gestosis and in 36 placentas of physiological pregnancies. The placentas from pregnancies with EPH-gestosis showed markedly higher activity of cathepsin B, no difference in the contents of SH-group, slightly higher activity of cystatins and they contain less deoxyribonucleic acids than the placentas from physiological pregnancies. The obtained results show that proteolytic--anti-proteolytic balance in placentas from pregnacies with EPH-gestosis is changed to the advantage of cathepsin B. This protease may in formation of structural and functional changes observed in placentas during EPH gestosis. PMID- 11387993 TI - [Coexistence of female sexual organ malformation and urinary tract anomalies]. AB - OBJECTIVES: The aim of the study was to estimate the correlation between sexual organs and urinary tract malformation. MATERIALS AND METHODS: The retrospective analysis of clinical data obtained from 50 patients with sexual organs anomalies diagnosed in the Academic Gynaecological Centre between 1992-1999 was performed. RESULTS: Material included 24 patients with the Meyer-Rokitansky-Kuster-Hauser syndrome and 26 with other sexual organs' malformations. Frequency of urinary tract anomalies was 42%, 11 patients with Meyer-Rokitansky-Kuster-Hauser syndrome and 11 with other malformations. The gravity of urinary tract anomalies showed no statistical significant difference dependent on the type and symmetry of genital malformation. Urinary tract anomalies were more frequent in cases of asymmetric genital malformation and the difference was statistically significant. CONCLUSIONS: Intravenous urography proofed to be more sensitive than ultrasonography in diagnosing urinary tract anomalies. PMID- 11387994 TI - [Peroxidation components of sperm lipid membranes in male infertility]. AB - We have studied the lipid peroxidation product (malondiadehyde-MDA) levels after thiobarbituric acid reaction, spectrophotometrically and by a high-performance liquid chromatography (HPLC) with UV detection in seminal plasma as well as in the cell fraction. Semen samples were obtained from healthy volunteers and infertile males. Ejaculates were previously classified into studied subgroups according to standard andrological criteria (sperm number, motility, morphology) and defined as: normozoospermia (K), azoospermia (Az), teratozoospermia (T), asthenoteratozoospermia (AT), oligoasthenoteratozoospermia (OAT) and idiopathic infertility (NIF). MDA is a good marker for lipid peroxidation. There were found elevated MDA concentrations (determined by HPLC) in seminal plasma of the all analysed pathological semen samples, especially in patients with OAT. The more pronounced differences between healthy controls and NIF patients were observed in intracellular compartment. The lipid peroxidation of rich in unsaturated fatty acids sperm membranes is considered to be one of the most important effects from ROS-induced cell damage what may lead to persistent infertility. PMID- 11387995 TI - [Fimbria form of primary fallopian tube carcinoma--case report and literature review]. AB - The classification of primary fallopian tube cancer (PRJ) set in 1991 in accordance to FIGO is unquestionable the best prognostic marker. Nowadays there some circumstances indicate the necessity of its modification. Authors describe a clinical course and morphological picture of PRJ located in fimbria. The subclinical form of PRJ was diagnosed during an explorative laparotomy due to high blood CA 125 level. In the modified classification of PRJ the fimbrial localization of cancer is defined as IF grade. We think, that in this special form of PRJ there is considerable possibility of early metastases to ovary and to the peritoneum. Besides because of fimbria peculiar anatomic structure the cases of PRJ located in fimbria should be classified as IIF grade. The role of blood CA 125 as PRJ marker is then emphasized. PMID- 11387996 TI - [Treatment of intracranial aneurysms during pregnancy]. AB - Aneurysmal subarachnoid hemorrhage (SAH) during pregnancy is rare but serious obstetric complication. SAH occurs more often in primiparae and in the third trimester of pregnancy. The hemodynamic and endocrine changes play an important role in the growth and rupture of aneurysms. There are no differences in the clinical course of SAH among pregnant and non-pregnant patients. Subarachnoid hemorrhage during pregnancy may be confused with eclampsia. The confirmation of the diagnosis is made by computed tomography (CT) or lumbar puncture and cerebral angiography. Ruptured intracranial aneurysms should be treated as they would be in patients who are not pregnant. When the aneurysm is successfully clipped, the pregnancy can be allowed to progress to term. In these cases the vaginal delivery is preferred by most clinicians. Caesarean section would be indicated in several circumstances: when the clinical state of mother is severe (coma, brain stem damage), if the aneurysm is diagnosed at term of labour, if the interval between the neurosurgical treatment of aneurysm and labour is less than 8 days. PMID- 11387997 TI - [Teratology, 40 years after the thalidomide tragedy]. AB - The article describes present knowledge of teratogenicity, which final manifestation are functional disorders, growth retardation, fetal carcinogenesis, malformation and death. PMID- 11387998 TI - [Ectopic pregnancy. I. Diagnostic-therapeutic algorithm used in clinics at the Gynecology Department of the Polish Mother's Memorial Institute]. AB - In prospective study from 1988 to 1992 I analysed possibility of early non invasive detection and treatment of ectopic pregnancy. In this purpose I used a gynecological examination, estimation of concentration of beta-hCG and progesterone in serum, transvaginal ultrasound and in certain patients, curettage. The fusion of all these methods permits to gain right diagnosis of ectopic pregnancy. In this work I described diagnostic and therapeutic algorithm of ectopic pregnancy allows in using conservative treatment in this pathology. Empty uterus (without gestational sac) in transvaginal ultrasound examination, in cases with beta-hCG concentration is higher than 3000 mIU/ml, allows recognise ectopic pregnancy. I introduced predictive scale for possibilities of conservative treatment and therapeutic algorithm allows to chose way of treatment (laparatomy, laparoscopy, Methotexate i.m. expectant management). The estimation of clinical value of diagnostic algorithm for ectopic pregnancy will present in part II and efficacy of conservative treatment methods in patients with ectopic pregnancy in part III. PMID- 11387999 TI - [Ectopic pregnancy. III. Estimation of efficacy of conservative treatment methods]. AB - From 162 patients with recognised ectopic pregnancy, 75 was qualified to conservative treatment. We estimated efficacy of therapy in 63 women treated by administration 50 mg/m2 i.m., of Methotrexate. For 50 of them therapy was successful (79.4%), another 13 (20.6%) demanded surgical procedures(laparatomy or laparoscopy), remained 12 was qualified to expectant management. Efficacy, in treated by Methotrexate group, significantly increasing to 86.8% (46/53) in patients with initial beta-hCG concentration in serum lower than 10,000 mIU/ml. PMID- 11388000 TI - [Microheterogeneity of two acute phase proteins in patients with ovarian carcinoma]. AB - Concentrations of two acute phase proteins: alpha 2-macroglobulin (alpha 2M) and transferrin (Tf) as well as glycosylation profiles of alpha 2M and Tf were studied in sera samples from 13 patients suffering from ovarian cancer. In the observed group of patients with ovarian cancer in whom the progression of disease was noticed low concentrations of both investigated proteins were present. On the contrary, the microheterogeneity of both alpha 2M and Tf was changed towards variants of both proteins more weakly reacting with ConA, what was previously described for AGP and ACT in all chronic inflammatory conditions including cancer. PMID- 11388001 TI - [A rare case of non-Hodgkin's lymphoma of the uterine cervix]. AB - We report a rare case of primary extranodal non-Hodgkin lymphoma of the uterine cervix. A 58-year old, postmenopausal multipara was admitted to the hospital with metrorrhagia and vaginal discharge. After diagnostic biopsy, the patient underwent radical abdominal hysterectomy, bilateral salpigo-oophorectomy and pelvic lymph node dissection. histologically, the lymphoma was classified as non Hodgkin lymphoma of intermediate malignancy. Immunohistochemical studies revealed that the atypical lymphoid cells were B-cells (CD 20+), indicating that the lymphoma was of B-lineage. Postoperatively, she received adjuvant chemiotherapy. Our experience and a review of current literature are presented. PMID- 11388002 TI - [Electroejaculation--effectiveness in a patient after retroperitoneal lymph node dissection--case report]. AB - Semen was obtained by transrectal electrostimulation from male patient after retroperitoneal lymph node dissection who had not ejaculated since time of operation. Vibratory stimulation was unsuccessful. electroejaculation was done under general anaesthesia. A silicone catheter was used to tamponade the bladder neck and antegrade ejaculations were collected. The semen underwent seminological assessment and then was diluted 1:1 in egg yolk substrate containing 15% glycerol, equilibrated, divided into straws and frozen in nitrogen. The straws were then placed for storage in liquid nitrogen until the insemination. PMID- 11388003 TI - [Assisted reproductive medicine in the next age]. AB - Medicine is going from success to success, but with the ever-increasing expectations of the society, the more it achieves, the less it satisfies. This is also true for reproductive medicine. Indeed, advances in basic research and the practical applications of the research that have been performed since the first successful in vitro fertilisation (IVF) in 1978, have given rise to hitherto unimaginable strategies for achieving human reproduction. Couples, that were considered sterile only a few years ago, can now acquire assistance conceiving. Twenty two years after Louise Brown was born, the reproductive medicine methods have spread all over the world and thousands of children have been born as a result of these techniques. ICSI applied to epididymal or testicular sperm in cases of azoospermia permits infertile men to become fathers. Even if the results of reproductive medicine have improved in terms of numbers of pregnancies, it is still striking that it is necessary to use stimulation which sometimes leads to hyperstimulation and multiple pregnancies, that embryo development in vitro is still limited, that implantation only occurs for 15-20% of embryos and this ratio have not changed in the last 20 years. The pregnancy rates currently obtained with IVF are at the most similar to those occurring in vivo. We still need to improve techniques to gain pregnancy rates approaching 50% per embryo. In order to fullfil this task, we need to introduce less aggressive and much simpler ovarian stimulation, routine culture of embryos to the blastocyst stage, pre implantation genetic diagnosis, cryopreservation and in vitro maturation of oocytes and finally to improve the implantation rates by our ability to understand and control the dialog between blastocyst and receptive endometrium. PMID- 11388004 TI - [Ectopic pregnancy. II. Evaluation of the clinical value of a diagnostic algorithm for ectopic pregnancy]. AB - In this work we assessed clinical value of diagnostic algorithm of ectopic pregnancy introduced in the first part of study. We calculated sensitivity, specificity, exactness and positive and negative predictive value. Studies embraced 374 women admitted to Polish Mother's Memorial Institute in Lodz with suspicion of ectopic pregnancy. In 143 patients we recognised ectopic pregnancy in remained 231 absence of pregnancy, intrauterine pregnancy or abortion. Diagnostic algorithm, with estimation hCG and progesterone concentration in serum and use transvaginal ultrasound, seems to be useful tool allowed diagnosed early ectopic pregnancy. We obtained 100% sensitivity and specificity of algorithm in diagnosis ectopic pregnancy. PMID- 11388005 TI - [Primary surgical treatment of congenital club feet]. AB - On the basis of available literature and his own experience the author presents some factors which influence the decision-making process in surgical treatment of congenital club foot and factors influencing the results of such treatment. The child's age at the time of operation, the type of the deformity after non operative treatment, and the authors' indications for surgical treatment are considered. PMID- 11388006 TI - [Long-term results of surgical treatment for clubfoot with the posterior release method]. AB - The authors present long term results of congenital clubfoot by posterior release. Our material consisted of 57 patients, with 57 clubfeet, 5 to 27 months old (mean age 9.5 months) at the time surgery. The age at the time of the final follow-up ranged from 8 to 20 years (mean age 14.7 years). The time of the observation ranged from 7 to 20 years (mean 13.9 years). Final results were evaluated according to the Magone classification. Basing on this classification we achieved very good results in 21.3%, good results in 29.3%, sufficient results in 26.7% and poor results in 22.6%. In the authors opinion posterior release and Achilles tendon elongation give the best possibility of correction of residual deformation. Varus deformity, abduction and equinal position should not be treated by posterior release. PMID- 11388007 TI - [Surgical treatment of congenital clubfoot with constriction band syndrome]. AB - Twelve feet in 11 patients with constriction band syndrome age ranging from 3 to 18 months (mean age 11 months) were evaluated in this study. The constriction band was located at the in the calf, at distal, media and proximal third levels. All these patients were treated surgically by a wide peritalar release using the Cincinnati incision, followed by a two-stage Z-skin plasty. Both clinical and radiological results were evaluated. In our material an excellent result was achieved in 1 case, good results in 3 cases, satisfactory in 4 cases and poor in the remaining four cases. Very good and good results were achieved in feet with shallow bands type II and III, while poor results were found in feet with deep constriction band type I. Early surgical treatment followed by serial casting leads to better functional results. PMID- 11388008 TI - [Analysis of intraoperative complications during surgery for clubfoot]. AB - The paper presents an analysis of the complications accompanying surgical treatment of congenital club feet. In the year 1966-1993 178 feet in 149 patients aged ranging from 5 to 140 months (mean age 17.5 months. In 57 cases a posterior capsulotomy with lengthening of the Achilles tendon was performed, in 121 cases a total sub-talar release was performed. Complications wee classified as intra operative and early and late post-operative. A total of 85 complications in 44 patients were observed, which amount to 44.7% of the cases. In 16 cases intra operative complications were a result of technical mistakes during the procedure (severing of a tendon, blood vessel or joint). Post-op complications were due to wound healing disturbances in 28 cases and necrotic changes in 12 cases. Sub talar release gives a larger number of complications (55.4%) compared to posterior capsulotomy with Achilles tendon lengthening (31.6%). PMID- 11388009 TI - [Evaluation of donor site morbidity after radial flap harvesting]. AB - Donor sites of 58 radial forearm flaps in 56 patients were analysed and evaluated on the basis of documentation and follow-up examination. In 2 patients 2 radial flaps were harvested. There were 25 females and 31 males aged 9-85 (mean age: 35 years). 35 free flaps (31 on to feet, 2 on to tibial stumps and 2 on to contralateral forearm) and 4 pedicled flaps (on to the head) were transferred to distal body regions. Two of them included a part of the vascularized radius. The same upper extremity was covered with 19 pedicled radial forearm flaps (13 with retrograde blood flow). Six of them were osteo-fascio-cutaneous flaps. After harvesting 52 flaps (in 50 patients), the donor sites were covered with full or split thickness skin grafts. In 5 cases the donor sites were sutured, and in one case local V-Y plasty was used. In 11 cases the following complications were observed at the donor sites: delayed healing of the wound for more than 6 weeks in 4 cases;--chronic vascular insufficiency in 1 case;--dysesthesias on the dorsal part of the hand in 3 cases;--periodical pain and oedema of the hand in 2 cases;--unacceptable hypertrophic scar in 1 case. No acute ischemia of the hand, no fracture of the radial bone, and no neuroma of the sensory branch of the radial nerve were observed at the donor site. PMID- 11388010 TI - [Use of pedicled cutaneous groin flaps in distal reconstruction of the upper extremity]. AB - In the years 1981-1997 at the Department of Plastic Surgery Medical Centre for Postsgraduate Education in Warsaw 116 patients were treated surgically using groin flaps and 97 patients using a pedicled cutaneous groin flap. This paper reports the results of the later technique. Reconstructions were performed in 10 female patients aged 18-58 (mean age 37.5 years) and 87 male patients aged 15-67 (mean age 33.8 years). The tissue defects or acquired deformations were caused by: crush injuries (26 cases), scalping injuries (23 cases), rugged injuries (18 cases), avulsion trauma (15 cases), explosion injuries (8 cases) and electric burns (7 cases). Flap size depended upon extent of the tissue defect and the from flaps were 7-26 cm long and 4-12 cm wide. Flap area ranged from 35 to 260 square centimetres. Emergency procedures were performed in 59 patients (61%). Secondary reconstructions were carried out in 38 cases (39%). Operative technique was based on the rules described by McGregor and Jackson. The donor site was sutured primarily tubulizing its basis--as in tube flap. The flap pedicle was cut off during a one stage procedure in 41 patients 21-30 days (mean 23 days) after surgery or during a two-stage procedure in 56 cases. The two-stage procedure consisted of an incision of part of the pedicle after 15-45 days post-op (mean 21 days) followed by a complete dissection after a few days (mean 4 days). In 44 cases the flap required modelling i.e. excision of excessives kin and/or thinning of subcutaneous tissue. The flap healed in 96 patients (99%). Complications at different stages of the treatment were observed in 40 patients (41%). In 3 cases wound ischemia was observed because of too tight suturing. Removal of skin sutures lead to normalization of blood supply. In 7 patients cyanotic skin of the distal part with no significant consequences was observed. In 27 patients (28%) necrosis of the marginal tissues surrounding the operation wound after cutting of the pedicle was noted. Besides supported necrectomy a conservative treatment was also applied by putting dressings soaked with antiseptics and 0.5% neomycine solution on a daily basis. This prolonged treatment by further several weeks. No significant complications were observed at the donor site. PMID- 11388011 TI - [Tests for evaluating various methods of bone fixation in digital replantation]. AB - Clinical results comparing different methods of bony fixation in 50 replanted digits in a series of 41 patients over a 13 year period are presented. The analysis was restricted only to replantantions involving sharp amputations of the proximal phalanx in order to restrict this study to a relatively homogenous group of patients. Evaluated techniques included: single and crossed Kirschner wire, intraosseous wire with or without Kirschner wire support, intramedullary splintage and intramedullary screws. Angulation deformities and delayed union were most common in cases treated with Kirschner wire technique. Nonunion was noted in 1 case (2%) stabilized with a single Kirschner wire. Angulation deformities were noted in 6 cases (12%). No such complications were noted in cases treated with intramedullary splintage and intramedullary screws. The best stabilizing effect and the most rapid union was noted in cases treated with intramedullary screws. PMID- 11388012 TI - [Results of treating septic non-unions of the tibia with vascularized bone grafts and the Zespol stabilizer]. AB - The authors present their experiences with various applications of the Zespol stabilizer and different kinds of vascularized bone transfers used in the treatment of septic non-unions of the tibia. The clinical material consisted of 24 patients with infections of the lower extremities treated with different vascularized bone transplants. The Zespol method was used in 23 cases. Compression osteosynthesis was applied every time with the stabilizer placed over the skin. The necessity of mounting the stabilizer far from the bone did not influence the stability of the osteosynthesis. Depending on the length of the transplant single or multiple plare supports were used. Additional plaster cast was applied only during wound healing. No loosening of the stabilizer was observed. The importance of early weight bearing in order to achieve bone union and remodelling of the bone graft is strongly emphasized by the authors. The described treatment allows bone union within 8 months on average. The advantages of the Zespol stabilizer in treatment of complicated disturbances of bone union are presented. PMID- 11388013 TI - [The possibility of isolation, culture and storage of articular cartilage cells]. AB - Lesions of articular cartilage are a common problem and concern millions of people world-wide. A decrease in physical activity and pain symptoms among patients resulting from damage to articular cartilage have prompted research concerning new methods allowing cartilage regeneration. State-of-the-art treatment of articular damage depends very much on genetic engineering techniques. The aim of this paper was to determine the authors' own way of isolation, proliferation and storage of chondrocytes of articular cartilage. The material consisted of 30 rabbits, from which fragments of articular cartilage were taken. The study consisted of the following stages: isolation, chondrocyte proliferation, cell and matrix identification, storage and MTT tests. Matrix digestion was achieved using the following solutions: 0.1% type IA collagenase; 0.025% trypsin, a mixture of collagenase and trypsin. The greatest amount of cells were found after digestion of the basic matter of cartilage by 0.1% solution of type IA collagenase. When ascorbic acid was added to the medium, a 25% increase in cellularity was observed. A cumulation of procollagen mRNA was noted in the isolated cells. After about 21 days the isolated cells formed a multilayer structure, with the space between the cells filled with a substance that showed typical traits for cartilage matrix. Storing the isolated cells for less than 48 hours at room temperature gave a 90% survival rate. Most cells died after less than 12 hours when stored at 4 degrees C. The described method of chondrocyte isolation proved to be effective in preparing material for treatment of articular cartilage lesion. PMID- 11388014 TI - [Images of growth zones of the femoral head, neck and greater trochanter in MR examination]. AB - The possibilities of MR imaging were evaluated in 36 children with 58 normal hips. The analysed children were divided into 3 groups, basing on age at the time of investigation i.e. children less than 5 years old, age ranging from 5 to 12 years, and over 12 years of age. MRI allowed visualisation of all growth zones of the proximal femur. On T-1 weighted images the growth cartilage of the femoral head, neck, and greater trochanter were of intermediate signal intensity. On sequences using fat saturation and on gradient echo images the cartilage was of high signal intensity. The growth zone of the subtrochanteric cartilage was of low signal intensity on T-1 weighted images and high signal intensity on sequences using fat saturation and on gradient echo images. T-1 weighted images together with FLASH 3D FAT SAT seem to be sufficient for the analysis of the growth zones of the proximal femur. PMID- 11388015 TI - [Efficacy of operative treatment for pathological fractures in bone metastases in relation to length and comfort of survival]. AB - The paper presents a retrospective evaluation of 47 patients with bone metastases treated surgically during the last 10 years at our ward. The mean age of the patients was 62.5 years. There were 31 females (mean age: 62.8 years) and 16 males (mean age: 62.3 years). In 37 cases (78.8%) it as possible to establish the primary localization of the tumour: breast carcinoma--16 cases, ovary cancer 5 cases, lung cancer--5 cases, prostate cancer--5 cases, kidney cancer--2 cases, stomach cancer--1 case, vagina cancer--1 case, hepatocarcinoma--1 cases and plasmocytoma--1 cases. In 10 cases (21.1%) we were unable to establish the primary focus of the tumour. The localization of the metastases was as follows: femur--32 cases, humerus--6 cases, tibia--3 cases, lumbar spine--1 case. Patients treated very briefly after qualification for surgery, in some cases during emergency service. In 2 cases of metastases to the tibia amputations at the femur were performed. The remaining patients were treated by local excisions of the metastatic tumours, followed by: in 33 cases internal osteosynthesis and bone cement application; in 7 cases osteosynthesis, in 4 cases hip arthroplasties and posterior spine instrumentation in 1 case. In 6.4% we had poor results because of the death of 3 patients. The mean follow-up was three months. In 93.6% we had good and very good results--no pain, good function and independence during daily activities. Mean survival time was 13.5 month (range 5-28 months). PMID- 11388016 TI - [Characteristics of intra-articular injuries of the knee joint in coal miners]. AB - This study presents an analysis of chronic knee injuries among coalminers. The aim of this paper was to individuate a relationship between the work conditions in coal mines and the observed pathological changes in the knee joint, as well as to assess the results of treatment. A population of 120 coal miners age ranging from 20 to 46 years (mean age 33.4 years) working in conditions continuously stressing the knee joints was evaluated. It was found that the lower the seam where the miners worked, the greater was the damage to the cartilage. Meniscus injuries were found mainly in miners working in higher seams with smaller load on the knew. It was also found that in this population multiple structures of the knee were injured simultaneously. PMID- 11388017 TI - [Comparative evaluation of stress patterns in models of the normal hip joint and after implantation of the Parhofer-Monch type hip prosthesis]. AB - The paper presents a comparative appraisal of stress patterns of the normal acetabulum of the hip joint and after implantation of a Parhofer-Monch type prosthesis based on a computer simulation analysis. Three-dimensional physical models were constructed basing on finite element methods and loaded according to Pauwels' scheme with a force R = 2878 N. Mathematical analysis was performed on a computer equipped with a Pentium II processor, using professional software written for theory of elasticity problem solving. Equipotential stress patterns were designated according to Hubers' hypothesis. A redistribution and decrease in stress values in the bone directly around the acetabular component of the prosthesis was noted. PMID- 11388018 TI - [Analysis of quality of life in persons after arm amputations]. AB - This study presents an analysis and an evaluation of the quality of life in patients after amputations of the upper extremities, basing on functional anamnesis, tests and a questionnaire about daily activity. Patients after unilateral upper extremity amputations are more proficient at both daily activities and two-handed activities than patients after amputations of both extremities. Patients with amputations below the elbow joint were found to be more proficient at two-handed activities compared to patients with amputations above the elbow joint. Daily activities with performed with the same degree of ease by both patients with amputations below the elbow joint and above the elbow. This proves that that the elbow joint maybe functionally replaced relatively easily. Patients with ambilateral amputations at arm level were more proficient with daily activities than two-handed activities. PMID- 11388019 TI - [Traumatic hip dislocation complicated by soft tissue interposition--a case report and review of the literature]. AB - Traumatic hip dislocation is by far less common in children than in adults. It occurs 4 times more frequently in boys than in girls. The force of the trauma leading to hip dislocation can be surprisingly small. Incomplete reduction is mainly caused by soft-tissue interposition or fragments of cartilage from the acetabulum. In such cases an X-ray examination will reveal a lateral protrusion of the femoral head. CT scans and MRI investigation play an important role in further diagnostics of the patient. The paper presents the case of a 5 year old boy with traumatic hip dislocation complicated by soft-tissue interposition. After an unsuccessful attempt of closed reduction, the hip was reduced surgically. The interposing element was removed--part of the cartilagenous insertion (size: 3 x 3 mm) of the femoral head ligament. During a follow-up examination 10 months post-op no signs of necrosis nor loss of ROM were noted. PMID- 11388020 TI - [Morphogenesis of beta 2m (A beta 2m) amyloid deposits in dialysis patients]. AB - beta 2m related amyloidosis (A beta 2m) is a late complication of renal replacement therapy (RRT), first identified in 1980. It is preferentially located in the joints. Its clinical manifestations and risk factors have been clearly delineated. Still, its true prevalence and the factors determining the fibrillar precipitation of beta 2m remain to be defined. A prospective autopsy study of the joints of dialyzed patients, confirms that A beta 2m prevalence increases with dialysis duration and age of the patients at the onset of RRT. Prevalence is not significantly different according to the modality of RRT, hemo--or peritoneal dialysis. Of great interest is the finding that histologic evidence of A beta 2m preceeds by over five years the onset of clinical symptoms. The detailed study of A beta 2m deposits has allowed their classification into 3 successives stages: stage 1, impregnation of the cartilage; stage 2, extension to capsules and synovia; stage 3, invasion of synovial and capsular deposits by macrophages. In the first 2 stages, in the absence of macrophages, there is no immunohistochemical evidence of advanced glycation of the A beta 2m. We conclude that neither macrophages nor the advanced glycation of beta 2m are prerequisities of A beta 2m formation. PMID- 11388021 TI - [Gulf syndrome... Balkans syndrome...chronic fatigue syndrome]. PMID- 11388022 TI - [Breast implants: state of the controversy]. AB - The controversy that occurred during the 1990 years concerning the breast implants has revealed several new problems, related to comfort surgery. Considering that this surgery has developed considerably, we found useful to outline the difficulties and the drifts that appeared with the breast implants. They were present at all levels: patients, implant factories, judges and lawyers, media, certified laboratories. An increased vigilance was found to be necessary in order to provide the safety and the quality of this type of surgery. PMID- 11388023 TI - [Beta 3 adrenergic receptor: physiologic role and potential therapeutic applications]. AB - The characterization in 1989 of the gene encoding the beta 3-adrenoceptor helped to interpret the results of pharmacological experiments on atypical effects of catecholamines distinct from the classical activation of beta 1 and beta 2 adrenoceptors. In rodents, the beta 3 adrenoceptor is abundantly expressed in white adipose tissue where energy is stored in the form of triglycerides and in brown adipose tissue that is specialized for thermogenesis. Treatment of rodents with beta 3 adrenoceptor agonists induces a weight loss related to the stimulation of lipolysis in the two types of tissues. These results led to propose the use of these agonists for the treatment of human obesity and NIDDM. However, the poor lipolytic effect of these agonists in human adipose tissue and the recent discovery of functional beta 3 adrenoceptors in the human heart raise new questions on the therapeutic use of beta 3-adrenoceptor agonists in man. In the human ventricle, these agonists induce a negative inotropic effect. In vessels, stimulation of beta 3-adrenoceptors produces a vasodilation. If these effects are conserved in the failing heart, they could shed a new light on the pathogenic role of the hyperadrenergism associated with cardiac failure, as well as on its treatment with beta-adrenoceptor blockers. PMID- 11388025 TI - [Screening for hemoglobinopathies in medical practice: audit of gynecologists, pediatricians and generalists in Brussels]. AB - The haemoglobin disorders are frequent genetic diseases in tropical regions and in the mediterranean basin. They include thalassaemia and sickle cell disease. Because of important migrations of populations after the Second World War many big cities are confronted with the disease in West Europe nowadays. In Brussels where a neonatal screening has been organised in the majority of the maternity's, 45% of newborns have at least one parent originating from a region at risk for an haemoglobin disorder. One neonate among 2.000 is diagnosed with a major haemoglobinopathy at the screening. This rate is important and it appeared essential to evaluate the knowledge and need for complementary information among the doctors confronted with the disease. A survey was done in January 1999 and a questionnaire was sent to all gynaecologists, paediatricians and general practitioners in Brussels. The main results were a general self-evaluation of poor knowledge and a great need for complementary information. This survey was important to obtain adequate support of official health authorities in term of screening, prevention and financial aid to information campaigns. A consensus is needed among the different doctors for the best care possible of sick patients. PMID- 11388024 TI - [The coxibs, third generation anti-inflammatories]. AB - The coxibs, specific anti-Cox-2, represent an interesting evolution for the treatment of patients suffering from rheumatic conditions and presenting gastrointestinal ulcer risk. Anti-inflammatory effectiveness is comparable with the other NSAIDs, but the toxicity for the gastrointestinal mucosa is slightly lower. Contra-indications, extradigestive adverse affects and interactions are the same as for any other NSAIDs. Their risks and cost must be taken into account as such as the rheumatic condition of the patient. They may be prescribed to patients presenting a gastrointestinal risk and the relevant cost is reasonable. Pharmacovigilance survey at mid to long term is necessary. PMID- 11388026 TI - [Immune deficiencies: diagnosis, management, some perspectives]. AB - Severe primary immunodeficiencies (PID) are rare; their global incidence is comparable to that of childhood leukemia; they include more than 100 different entities. Clinical manifestations are: unusually severe or frequent infections or infections that do not respond to adequate treatment; an increased risk of certain malignancies; sometimes auto-immune manifestations. Delayed diagnosis and management of PID can lead to severe and irreversible complications or to death. PID can become manifest only in the adult; in common variable immune deficiency, the median age at diagnosis is between the 2nd and the 3rd decade of life. PID are often transmitted genetically; recent progresses in molecular biology have allowed more precise and earlier, including antenatal, diagnosis. Molecular treatment of 3 infants with a severe immunodeficiency has recently been achieved in April 2000. Those progresses were mostly based on the study of immunodeficiency databases. We present here the work of a Belgian group specialized in PID; meetings have started in June 1997. This group establishes guidelines for the diagnosis and treatment of PID, adapted to the local situation. The elaboration of a national register of PID is also underway; this has to provide all guaranties of anonymity to patients and families. Such a register already exists at the European level; it has provided the basis for new diagnostic and therapeutic possibilities. The inclusion of Belgian data in this register should allow essential progresses essential for our patients. PMID- 11388027 TI - [HIV infection and hepatitis B co-infection: survey of prevalence in pregnant women in Bobo Dioulasso, Burkina Faso]. AB - We carried out this study to assess the prevalence of co-infection with hepatitis B and HIV-1 in african pregnant women. Nine hundred and seventeen pregnant women attending two antenatal clinics in Bobo Dioulasso, Burkina Faso, were included. The characteristics of the women were the following: a mean age of 26 years and 83.5% married; a mean gravidity and para of 4 and 3 respectively and a mean gestational age of 27 weeks of amenorrhea. Then sera were drawn to be tested for both hepatitis B and HIV-1. Sera tested positive for HIV-1 at ELISA were confirmed by Western Blot. Ninety eight women (10.7%) were Ag HBs carriers of whose 18.2% were positive for Ag HBe, 66.7% anti-HBe positive and 95.6% anti-HBc positive. The HIV-1 prevalence rate was 5.9%. Eight women were positive for both Ag HBs and HIV-1, giving a co-infection rate of 0.88%. The co-infection rate by hepatitis B and HIV-1 is very low in pregnant women in Bobo Dioulasso despite a high prevalence of these two infections. PMID- 11388028 TI - [Can we prevent prostatic cancer: role of nutrition?]. AB - The present paper gives a comprehensive overview of recent data, especially prospective randomized trials, which support an important role for nutrition in the development of prostate cancer. Prostate cancer seems to be an ideal candidate for chemoprevention, in order to interfere by modification of nutritional habits with its onset, its incidence and ultimately with its progression, especially in high risk groups. PMID- 11388030 TI - [Workload and "poor use" of physicians]. PMID- 11388029 TI - [Terminal medical interventions: psychosocial, medical, ethical and legal aspects]. AB - We give an overview of the available medical solutions to help a patient with refractory symptoms at the end of his life. Patient "competence" must first be evaluated and, even if their diagnosis is difficult, organic mental disorders and depression must be diagnosed and adequately treated to allow a real, personal and honest dialog. Administration of high doses of morphine is frequently used at the end of life not only to fully relieve pain but also to accelerate death, even if this is not clearly stated. This technique is not devoid of hypocrisy and high doses of morphine can have quite unpleasant side effects. Treatment withdrawal or withholding is generally not sufficient to allow a correct end of life. The arrest of ventilation, dialysis, artificial nutrition and even more hydration must often be coupled with techniques inducing unconsciousness, which makes imprecise the limits between such a "passive" ending of life and "active" euthanasia. The technique of terminal sedation, frequently based on the use of midazolam, has been more recently introduced in some palliative care units. Such a "controlled sedation" is supposed to allow a "natural" death by inducing a profound sleep. In opposition with active euthanasia, which allows a quiet and rapid death at a moment chosen by the patient himself, this technique of "sedation" has an undetermined duration, has legal implications which could be viewed as quite similar as the ones of euthanasia, and, moreover, this prolonged agony can be extremely stressful and distressing for the family. Medical-assisted suicide is allowed in The Netherlands under the same conditions as euthanasia. Death is generally obtained after a few hours but the technique is not always successful and the process of death can sometimes be prolonged and uncomfortable. This technique can nevertheless be preferred by some physicians and patients. As compared to active euthanasia, the proportion of medically-assisted suicides (1/6) is low in The Netherlands. Euthanasia is the only technique able to induce a peaceful and rapid death. The proportion of various techniques to actively induce death is probably quite similar in our country than in The Netherlands but, most of the time, these interventions occur at the very end of life when the patient is no longer able to participate in the decision process and thus occur without his explicit request. We think that, as for all medical decisions, the use of one or the other of these various techniques should be selected after a quiet and free discussion between the patient and his physician, preferably in advance and not in a situation of emergency and panic. PMID- 11388031 TI - [J.E.P. 2000: commentary on "practical workshops" in cardiology]. PMID- 11388032 TI - [Idiopathic hypereosinophilic syndrome: from the clinic to the laboratory, from the laboratory to the clinic]. PMID- 11388034 TI - [From intensive care to the laboratory: biochemical modifications of the erythrocyte membrane glycoproteins in sepsis]. PMID- 11388033 TI - [Clinical value of the measure of global fibrinolytic activity]. PMID- 11388035 TI - [Lipoid nephrosis, the toad, the salamander and the earth worm. A nephrological exemplary tale]. PMID- 11388036 TI - [Technical realization of an LCD-based vision test device]. AB - We describe a computer-aided self-test vision screener for testing near and far visual acuity. The device generates images separately for each eye on a LCD. Acuity can be measured in the range between 0.1 and 1.6 for any distance between 0.34 m and 6 m. In addition, it also enables testing for colour deficiency, phoria, stereopsis and contrast sensitivity. The device is fully automatic and enables self-testing of the above mentioned functions. Initial practical application in an industrial environment has demonstrated the practicability of the device. PMID- 11388037 TI - [Use of powder metallurgy for development of implants of Co-Cr-Mo alloy powder]. AB - This paper discusses the application of powder metallurgy for the development of porous implantation materials. Powders obtained from Co-Cr-Mo alloy with different carbon content by water spraying and grinding, have been investigated. Cold pressing and rotary re-pressing methods were used for compressing the powder. It was found that the sintered materials obtained from water spraying have the most advantageous properties. PMID- 11388038 TI - Initial stability of modular acetabular components. Comparative in-vitro study with polyethylene and ceramic liners. AB - Modular acetabular components with alumina ceramic liners are currently used in total hip arthroplasty, but concerns have emerged regarding their high stiffness, which could cause impairment of stability, stress-shielding phenomena, and loosening. The purpose of the present biomechanical investigation was to compare the in-vitro initial stability of a modular press-fit acetabular component using a polyethylene liner and using an alumina liner. The initial stability was investigated by measuring the micromotion between the implant and the acetabulum during the application of physiological load (2.39 kN). The micromotion of the acetabular component was investigated in 10 acetabuli using a polyethylene liner and in 10 acetabuli using an alumina liner. Micromotion was assessed at the level of the Os ilium, Os pubis, and Os ischium using 3 electromagnetic transducers. The transducers have a sensitivity of 1 micron and a range of measurement of 500 microns. All implants have been fixed on human pelves made of polyurethane. Measurement of implant micromotion showed stable conditions at the level of the three main sectors of the acetabulum during all tests. No statistically significant differences of results were observed between the group of specimens with polyethylene liner and the group of specimens with alumina liner. The mean micromotion values of the uncemented cups were similar to the mean micromotion values of 10 cemented cups investigated to achieve comparative data of stability. In conclusion, the modular acetabular components inserted using an alumina liner showed a satisfactory initial stability in-vitro. The results do not contrast with those achieved using the same cup inserted with a polyethylene liner. PMID- 11388039 TI - [Pressure-dependent outflow resistance in cerebrospinal fluid dynamics: evaluation a calculation model for diagnosis of normal pressure hydrocephalus in an animal experiment with H-Tx rats]. AB - The internationally accepted methods of calculating cerebrospinal fluid dynamics proceed from the assumption of a pressure-independent resistance to CSF outflow. Our new model focusses on the pressure-dependency of this resistance. In it, we monitor the entire pressure course over time, p(t) during and after infusion. A comparison of the pressure rise, On(p), during infusion, and the decrease, Off(p), to the same pressure level, permits the creation of all the formulas for C(p) and R(p). The simultaneous measurement of resistance and compliance during a single intervention allows us to minimize patient exertion. In contrast to the classical methods, it is not necessary for the ICP to reach a plateau. Our mathematical model differs from the static examination model by describing a pressure-dependent slope of the function for the resistance. This has been demonstrated in a study using H-Tx rats. In this way, we are able to take the non linearity of the CSF resorption into consideration. PMID- 11388040 TI - Quantitative evaluation of the performance of different electrical tomography devices. AB - Two widely used electrical tomography systems, the Sheffield Mark I and the DAS 01P, were quantitatively evaluated and compared to the newly developed Goe-MF system. The performance was quantified using a hardware phantom which closely matches the real input and transfer impedances of the human thorax and allows measurements equivalent to different states of lung inflation. Our results demonstrate that adequate averaging is necessary for noise reduction for the Mark I and especially for the DAS-01P system to get meaningful results even in visualizing maximal respiratory manoeuvres. The Goe-MF system showed a notably improved signal-to-noise ratio which allows also dynamic measurements at low levels of lung volume changes, e.g., in intensive care lung injury patients. PMID- 11388041 TI - [Conditions and success factors of telemedicine innovation]. AB - Telemedicine and health telematics can play a significant role in improving the efficiency of health care systems. Currently, however, the potential of telemedical applications is far from being fully utilized. In the present study, an attempt has been made to identify factors that may aid the further dissemination and general acceptance of telemedical innovations. The results obtained suggest that apart from the factors under the sole control of legislation, and insurance providers, those that can be directly influenced by developers, manufacturers and providers of telemedical applications play an important role. Of particular importance for the successful and lasting use of such applications is the creation of promising business models, aspects of which are considered in the last part of the article. PMID- 11388042 TI - [Stem cells in medicine]. AB - We witness an unprecedented explosion of knowledge and interest in the field of stem cells and stem cell-dependent organ systems. The notion of stem cell has expanded to encompass cells with different properties but all amenable to ex vivo manipulation. We have learned that not only rapidly self-renewing tissues, but also unsuspected systems such as the central nervous system are associated with post-natal progenitor cells. The bone marrow seems to represent an important cross-road of many different progenitors, and perhaps the easiest source of diverse progenitors to be used for therapeutic purposes in a large spectrum of diseases. While some applicative uses of post-natal progenitor cells are either already in use, or soon to be routinely implemented, much larger perspectives are linked to the design of appropriate strategies for cell delivery and stable cell transduction. Meanwhile, major gains in understanding of the biology of different diseases are implied by the very recognition that many diseases are indeed stem cell diseases. PMID- 11388043 TI - [Perspectives of immunosuppressive therapy in kidney transplantation]. AB - Within recent years important progress has been made in the field of renal transplantation, both in terms of surgery and immunosuppression. At the present time renal transplantation offers a better expectation and quality of life to patients with end stage renal disease, compared with patients in dialysis. The survival of renal transplantation depends on many immunological and non immunological factors. The widespread use of calcineurine inhibitors (cyclosporine and tacrolimus) has improved the actuarial survival of transplanted kidneys at one year, decreasing the number and the severity of acute rejection episodes, but the calcineurine inhibitors did not improve the chronic rejection. The use of the new immunosuppressant drugs (mycophenolate mofetil, rapamicine and RAD, anti CD25 humanized antibodies) has contributed both to a further reduction of the number of rejection episodes and of side-effects of the immunosuppressant drugs, which have been possible thanks to the various combinations of these drugs. moreover these drugs allow the reduction or withdraw of steroids. Further studies will be necessary to ascertain that these new drugs improve the long term survival of transplanted kidneys and to define more precisely problems relating to the immunosuppressive therapy with respect to renal function, rejection, and patients' quality of life. PMID- 11388044 TI - [Professional licensing examination. Innovative experience: the Integrated Multimedia Pilot Project of the Chieti University]. AB - The Medical Licensing Examination (MLE) is governed, in Italy, by a law enacted in 1956. An ideal clinical competence assessment tool should effectively, reliably and objectively measure all the components of clinical competence: basic knowledge and clinical skills as history-taking, performing a physical examination, formulating the most likely diagnosis, establishing a management plan, communication and interpersonal relations. Since 1998 first session of the MLE, Chieti University implemented computer-based case simulations and standardized patients in the Multimedia Integrated Pilot Project (MIPP) administration. At the present time, we have examined 370 subjects during five sessions of MLE. This preliminary work shows results regarding the examinees in the first examination session of 1998 and 1999. The two groups of examinees are relatively homogeneous for number, age, gender, length of curriculum and country of University degree. In both groups the curriculum scores (preclinical, clinical and total) and the MIPP final score are reported for all subjects, first ten and last ten examinees. The MIPP final score is moderately correlated with the preclinical, clinical and total curriculum scores. Recently, the Federation of State Medical Boards and the National Board of Medical Examiners, sponsoring organisations of the United States Medical Licensing Examination (USMLE), have been planning the implementation of computer-based testing for the USMLE. It is important to point out that our MIPP-based MLE is not a mere didactic experiment, but a legal certifying examination valid for licensure. PMID- 11388045 TI - [Inflammatory aneurysm of the abdominal aorta. Study of 355 patients with aortic aneurysm]. AB - The Authors report their experience in the treatment of inflammatory aneurysm, on a whole of 355 patients consecutively operated on for abdominal aortic aneurysm. The incidence was 5.6% when relying on clinical aspects; 10.6% when considering the results of microscopy. All patients were operated on through a retroperitoneal approach: early and late results of surgery were not significantly different between the two groups of patients (inflammatory aneurysm 38, atherosclerotic aneurysm 317). PMID- 11388046 TI - [Lyell syndrome and antibiotic therapy]. AB - Lyell syndrome is an idiosyncratic reaction to drug treatment associated with high mortality due to difficulty in the diagnosis and lack of treatment with proven efficacy. We present the case of a patient treated with antibiotics who developed an exantema-like eruption, diagnosed as Lyell syndrome. The warning signs are represented by a diffuse exantema-like erythema generally associated with fever, large and soft bullae, resembling pemfigo, with subsequent transformation into diffuse erosions following detachment of the epidermis. Skin biopsy is decisive for a correct diagnosis. Negative direct and indirect immunostain, and negative Tzank cytodiagnostic test, associated with histologic findings consistent with "epidermic necrosis with diffuse vacuolar basal stratum degeneration", allow a rapid diagnosis of Lyell syndrome. In our patient, aggressive treatment did not obtain the positive results reported in the literature, possibly due to the advanced stage of the disease. PMID- 11388047 TI - [Hematopoietic stem cell transplantation in pediatric onco-hematology]. AB - Hematopoietic stem cell transplantation (HSCT) represents today an important therapeutic choice for several disorders of childhood: hematologic, metabolic and neoplastic pathologies can be treated with this strategy. The aim of this paper is to resume the latest history of HSCT, paying attention to the main changes and controversies, and its efficacy as far as concerns the major indications in pediatric oncohematology. PMID- 11388048 TI - [Genetic risk factors in ischemic cardiopathy]. AB - Coronary artery disease is a complex disease, characterized by a myriad of interactions between environmental and genetic factors. There is a growing interest about the genetic components. This research field is rapidly growing, and could offer new diagnostic and therapeutic tools in a near future. This paper will focus on common variations in several candidate genes. They have been categorized into three groups, according to different pathophysiological mechanisms: 1) lipid metabolism; 2) hemostatic balance; 3) non-lipid metabolism. PMID- 11388049 TI - [Epistaxis and circadian rhythm]. AB - It is known that many physiological variables show a rhythmic variability along a time span, e.g.: day, week, month or year. It is possible that the temporal overlapping of a constellation of factors may act as favouring and/or triggering factors for the onset of several acute diseases. Among these, epistaxis shows a circadian variability too. Such pattern, characterised by a prevalent peak in the morning and a secondary one in the evening, strictly resembles that of arterial blood pressure. The lack of differences between the temporal pattern of hypertensive and normotensive subjects suggests that the physiological pattern of blood pressure, and not strictly a condition of hypertension, may play a favouring role in certain hours of the day, probably together other local factors. PMID- 11388050 TI - [Inflammatory aneurysm of the abdominal aorta. Review of the literature of the past 30 years]. AB - A careful review of the available literature on the clinical and pathologic features of the so-called inflammatory aneurysm of the abdominal aorta is reported. The etiology of this peculiar disease is still obscure and controversial, even if the current trend is to define it as an immunomediate phenomenon consisting on an "exaggerated" inflammatory response to atherosclerosis. Suggestions about the similarity or identity from the pathologic viewpoint, of inflammatory aneurysm, periaortic fibrosis and idiopathic retroperitoneal fibrosis are attractive, but not universally accepted. The surgical treatment aims at the same objectives than for atherosclerotic aneurysms (i.e. prevention of rupture); operative management is however endangered by greater difficulties which hopefully may be reduced in the coming years by the use of corticosteroids and mainly through the wider application of endografting procedures. PMID- 11388051 TI - [Anisakiasis: a borderline disorder]. AB - Anisakis simplex is a nematode which can parasitize many different kinds of fish or cephalopods (codfish, salmon, tuna, mackerel, hake, etc). Anisakis simplex can cause different diseases in humans. The human being acquires the larvae by eating raw or undercooked seafood. Acute anisakiasis is probably caused by an inflammatory and/or allergic response in the digestive tract mucosa with abdominal pain. It can also induce IgE-mediated reactions with several clinical manifestations ranging from urticaria/angioedema to anaphylaxis. Chronic anisakiasis results from abscesses or eosinophilic granulomas caused by parasite invasion. This later form can mimic appendicitis, duodenal ulcer, inflammatory bowel diseases and intestinal obstruction. An early gastroduodenoscopy can confirm the diagnosis and prevent the complications. Serodiagnosis of anisakiasis is difficult since many Anisakis antigens show cross-reativity complications. In fact many people have high IgE titles in the absence of obvious allergic reactions to seafoods. As preventive measures heating for 10 min over 65 degrees C or freezing (minus 20 degrees for 24 h) destroys the infectivity of the larval stage but not always prevent allergic reactions. PMID- 11388052 TI - [Mammography screening in Germany]. PMID- 11388053 TI - [Concept and implementation of model projects for mammography screening in Germany]. AB - The carriers of the German Statutory Health Care System have recognized that only mammographic screening according to the European Guidelines for the Quality Assurance of Mammography Screening will permit early detection of breast cancer with an acceptable risk/benefit ratio. In the German pilot projects, regional mammography screening programmes according to the European guidelines are being tested in the framework of the German health care system. The European guidelines require comprehensive quality assurance of all relevant steps in the chain of events involved in screening, from invitation on to therapy and follow-up. Such comprehensive quality assurance involves several medical specialities and other professional groups dealing with out-patient and in-patient health care and requires long-term cooperation with public institutions (population registries, cancer registries). The objective of the pilot projects is to test the organizational and legal conditions essential to introduction of a mammographic screening programme according to the European quality assurance guidelines in Germany. PMID- 11388054 TI - [Development of the incidence and mortality of breast cancer]. AB - In Germany, breast cancer is still the most frequent cancer site with approximately 45.000 incident cases and 18.000 deaths annually. After a continuous increase of mortality for decades and a short period of stagnation, a decline can be observed since a couple of years. However, this turnaround has not been preceded by an analogous evolution of breast cancer incidence. A similar development can also be observed in other countries. The conditions under which this changing trend has occurred indicate that it might have been caused mainly by better treatment and only to a low extent by altered prevalence of risk factors or the implementation of screening programmes. Due to the expected latency, an influence of organized screening on mortality can be expected only at the end of the 1990s, i.e., at the end of the period for which data is available or later. In the discussion about implementation of quality-controlled screening in Germany, it should be kept in mind that the quality of the entire programme- starting with screening and ending with therapy--will decide to what extent breast cancer mortality can be reduced. PMID- 11388055 TI - [Evaluation of current mammography screening studies in Europe and North America]. AB - The ancient randomized-controlled trials of breast cancer screening have offered clear evidence that invitation to participate in mammography screening with/without clinical examination is effective in breast cancer mortality reduction und underline the detection sensitivity of mammography. Recently published criticism of both the effect and fundamental data handling could be shown to be untenable. Randomized studies underestimate the effect while well designed observational studies do not systematically overestimate the magnitude of the effect. To transform high-quality study results in all-day's practice is a hugh challenge, but affordable as has been demonstrated in Great-britain, the Netherlands and Sweden. PMID- 11388056 TI - [Plans of the German Roentgen Society and the Professional Association for Quality Assurance in Mammography]. AB - A high level of quality is an unequivocal prerequisite for obtaining the highest possible accuracy in symptomatic patients and for reproducing the results concerning mortality reduction, which were obtained in large screening trials. Present deficiencies in Germany are due to legal regulations, which have not been updated and which are thus below European standard. Furthermore the quality assurance program has not proven sufficiently effective for mammography. In order to promote mammographic quality assurance, the German Roentgen Society proposes an accreditation program. The accreditation, which concerns A.) mammographic technique and positioning and B.) mammographic reporting is not obligatory, but will allow acquisition of special official certificates, which may support the patients to find doctors who perform and read mammograms with high quality and expertise. The accreditation shall be performed by personnel and/or institutions who are specifically trained surveyed. PMID- 11388057 TI - [Value and significance of digital full-field mammography within the scope of mammography screening]. AB - To evaluate the digital technique for screening mammography various experimental studies and clinical examinations were performed by using direct full-field digital mammography (FFDM). The findings concerning the detectability and characterization of microcalcifications and soft tissue masses as well as the radiation exposure were compared to the state-of-the-art conventional screen-film mammography (SFM). The results of these studies revealed a high performance of the digital images, which are at least equivalent to the conventional images, whereas digital spot views were significantly superior to conventional ones particularly in the detection of microcalcifications. This was especially true, when the potential of post-processing was used. In addition, the sensitivity of FFDM should be increased, if computer-aided-diagnosis (CAD) is available. Furthermore, the patient radiation dose can be significantly reduced. Additional advantages are quick and easy handling, efficient data transfer and digital archiving. Thus, FFDM will become an important tool in screening mammography. PMID- 11388058 TI - [Prevention in familial breast cancer susceptibility]. AB - First retrospective data show that hereditary Breast cancer risk can be positively influenced by prophylactic surgical procedures and hereby a thorough consultation seem to be an option for mutant gene carriers. A first evaluation of the women of 1050 high-risk families by the German Consortium "Hereditary Breast and Ovarian Cancer promoted by the German Cancer Aid (Deutsche Krebshilfe)" has shown that approximately 80% of the women concerned decide to participate in an intensive early detection--programme and only 5% of the women have decided to submit themselves to prophylactic surgical measures. This again shows how absolutely necessary the elaboration and evaluation of early detection measures within the different investigation projects are. PMID- 11388059 TI - [Ultrasound mammography and magnetic resonance mammography as adjunctive methods in mammography screening]. AB - Ultrasonography and magnetic resonance (MR) tomography are helpful tools to evaluate unclear lesions found at screening mammography. Ultrasound is particularly useful to prove the presence of a cyst or to further examine unclear, asymmetrical densities. With MR mammography, carcinomas can be found even when x-ray mammography or ultrasonography are limited due to diffuse, benign, proliferative changes. Ultrasound guidance has greatly facilitated core needle biopsy for suspicious lesions. Additionally, approved devices for MR guided biopsy are commercially available. As a primary screening tool, ultrasound or MR mammography may be used only in women who are positive for BRCA-1 or BRCA-2 mutations or who are otherwise at high risk for breast cancer. PMID- 11388060 TI - [Mammography/stereotactically controlled vacuum excisional biopsy. Interventional methods in breast diagnosis]. AB - As well as in the complementary mamma diagnosis (clinical examination, mammography, sonography) including the dynamic MRI and the mammography screening projects the interventional methods like mammographically/stereotactically guided vacuumbiopsy/excisional biopsy are established methods to avoid unnecessary open operations and they are integrated in the clinical routine. By using strict criterias it is a way to improve the specificity and the negative predictive value in the mamma diagnosis. In addition the interventional methods requires less costs, less time and reduces the morbidity in comparison to an open operation. PMID- 11388061 TI - [Assessment of usefulness and risk of mammography screening with exclusive attention to radiation risk]. AB - In Germany, a vivid debate is still going on concerning the implantation of preventive mammographic screening into the national health care system. Before launching such a programme, benefits and risks have to be carefully assessed. The risk of radiation induced breast cancer was calculated in terms of mortality per 100.000 women using the risk estimates of BEIR. The radiation dose was assumed to be 8 mSv per examination (2 mGy per view, 2 views; RBE = 2, to be on the safe side). From the current literature regarding the benefits of screening, the number of deaths avoided per 100.000 women was estimated. Under the assumptions made in the model, screening from age 40 to 50 (six examinations at 2 years intervals, discontinuing at later age) might be associated with a benefit/risk ratio of about 6, whereas for women in the age group from 50 to 60 years the benefit/risk ratio was shown to be about 25. For a screening programme starting at the age of 50, the radiation risk involved does not appear to be a crucial issue. For performing mass screening with mammography beginning at the age of 40, radiation risks might have to be considered among a number of other risks associated with this age group. PMID- 11388063 TI - [Multiple cavernous infiltrates in both lungs. Pulmonary involvement in Wegener granulomatosis]. PMID- 11388062 TI - [Recurrence of cystic adventitial degeneration of the popliteal artery--magnetic resonance tomography and MR angiography]. AB - Adventitial cystic disease is a rare disorder characterized by mucin-containing cysts of the adventitial tissue. The condition has a predilection for the popliteal artery. Men are predominantly affected, usually around the fourth decade. We report of a case of recurrent adventitial cystic disease and the possibilities of modern cross-sectional imaging. In particular we discuss the advantages of magnetic resonance imaging and magnetic resonance angiography for the diagnosis of this condition. Furthermore, the etiology and the possibilities of surgical treatment are illustrated. PMID- 11388064 TI - [Diverticulosis and diverticulitis]. PMID- 11388065 TI - Research governance--global or local? PMID- 11388066 TI - Assisted reproductive techniques and the law. PMID- 11388067 TI - Errors of recall and credibility: can omissions and discrepancies in successive statements reasonably be said to undermine credibility of testimony? PMID- 11388068 TI - Expert witnesses and experimental treatment: the extent of the duty to warn of risks when using experimental treatment. PMID- 11388069 TI - Coroners. PMID- 11388070 TI - Advances in genetics: spiralling into trouble? PMID- 11388071 TI - Research governance, consent and evidence-based medicine. PMID- 11388072 TI - Shall I open a bargain basement? (an allegory). PMID- 11388073 TI - Health and safety: better or worse in public enterprise or private enterprise? PMID- 11388074 TI - Alec Samuels puts the case that the company doctor owes little, if any, duty of care to employees or prospective employees. PMID- 11388075 TI - [Blood is a special juice]. PMID- 11388076 TI - [Is there progress in chemotherapy for breast cancer?]. AB - Breast cancer is the most frequent type of cancer in women. The therapeutic approaches to breast cancer have developed rapidly over the past 20 years, due to the increasing knowledge about the biology of breast cancer. Therefore it was possible to increase response rates as well as the duration of response and survival in neoadjuvant, adjuvant, and palliative treatment. This paper gives a review of the current breast cancer trials. Effective new cytotoxic chemotherapy and hormonal therapy agents, as well as the identification of specific molecular abnormalities (HER2/neu) led to the development of targeted therapeutic interventions in the neoadjuvant, adjuvant, and palliative treatment of breast cancer. Increased understanding of the biology of breast cancer led to the development of rational therapeutic interventions, which are currently under active clinical development. PMID- 11388077 TI - A ten-year analysis of demographic trends for cutaneous melanoma: analysis of 2501 cases treated at the University Department of Dermatology in Vienna (1990 1999). AB - BACKGROUND: The aim of the study is to provide recent demographic data on cutaneous melanoma in Austria. PATIENTS AND METHODS: Patients with primary cutaneous melanoma diagnosed between January 1, 1990 and December 31, 1999 were included in the analysis. Data were retrieved from the melanoma registry of the University Department of Dermatology in Vienna. RESULTS: In the observation period, 2501 cutaneous melanomas were diagnosed in 2427 patients (mean age: 55.4 yrs; 51.2% females). A total of 267 (10.7%) cutaneous melanomas were in situ, and 2234 (89.3%) were invasive. The median invasion thickness according to Breslow for invasive melanomas was 0.98 mm (25%-75% percentiles: 0.55-2.00 mm). We observed a significant trend towards thicker tumors with increasing age (p < 0.001). In patients less than fifty years of age, a significantly higher Breslow thickness was observed in men than in women (0.90 mm versus 0.80 mm, p = 0.03). The median Breslow thickness of invasive melanoma decreased from 1.20 mm in 1990 to 0.92 mm in 1999 (p for trend, < 0.001). Compared to women, men had melanomas more frequently on the back (43.8% versus 24.4%, p < 0.001). In women, melanomas were more frequently found on the lower legs than in males (21.5% versus 6.7%, p < 0.001). In a multivariate model, invasion thickness according to Breslow was the single most important predictor of survival. CONCLUSION: Our data confirm recent reports from other European countries with regard to the decline in tumor thickness of cutaneous melanoma. Our data also demonstrate a need for improving early diagnosis, particularly in certain subgroups of patients. PMID- 11388078 TI - Serum 25-hydroxyvitamin D and parathyroid hormone in patients with ankylosing spondylitis before and after a three-week rehabilitation treatment at high altitude during winter and spring. AB - Does a sojourn at high altitude during the winter and spring improve vitamin D status (and possibly suppress parathyroid hormone [PTH]) in patients with ankylosing spondylitis (AS)? In 73 patients with AS, serum concentrations of 25 hydroxy-vitamin D [25(OH)D] and PTH were determined before and after a three-week rehabilitation treatment at Bad Gastein (1000 m above sea level). At the first examination, serum 25(OH)D was median (25th, 75th percentile) 15.5 ng mL-1 (10.0 ng mL-1, 20.6 ng mL-1). Thirteen patients (18%) had a 25(OH)D concentration below 8 ng mL-1. In 53 patients (73%) the level was below 20 ng mL-1. After the sojourn, 25(OH)D significantly (p = 0.02) increased to 19.7 (11.3, 24.6) ng mL-1. PTH did not change significantly, being 32 (22.4, 43.9) pg mL-1 before and 30.3 (24.1, 39.9) pg mL-1 after the sojourn. Analysing different periods of sojourn, a significant (p < 0.001) increase in 25(OH)D was found in April but not in the other months. Patients with ankylosing spondylitis may have extremely low levels of 25(OH)D. The results of the present study suggest that a sojourn at high altitude in early spring is liable to reduce vitamin D deficiency. PMID- 11388079 TI - [Relationship between symptoms of depression and anxiety and the quality of life in multiple sclerosis]. AB - BACKGROUND: Health-related Quality of Life (QoL) is gaining increasing influence as a relevant evaluation criterium in clinical research. Several studies have investigated QoL in patients suffering from multiple sclerosis (MS) and the impact of therapeutic interventions on QoL. However, the influence of psychiatric complications, which occur frequently in MS (e.g. depression and anxiety disorders), on the autoassessment of QoL are hardly ever considered. METHODS: Symptoms of depression and anxiety were rated in 74 outpatients with definite MS. The health-related QoL was assessed and set into relation to physical disability (measured with the Expanded Disability Status Scale) and duration of the disease. The results were compared with 74 normal controls of the same age. RESULTS: A highly significant relationship between emotional state (Zerssen-scale), depression (Zung-depression-scale), anxiety (Zung anxiety scale) and Quality of Life was evident. CONCLUSIONS: Clinical trials assessing Quality of Life in MS patients should consider the frequency of psychiatric comorbidity and the influence of depression and anxiety disorders on self-rated Quality of Life. PMID- 11388080 TI - Insulinoma induced hypoglycemia in a type 2 diabetic patient. AB - Insulinoma in a patient with pre-existing diabetes mellitus is extremely rare. We report a case of an insulinoma in a patient with type 2 diabetes mellitus who after 14 years of sulfonylurea treatment experienced recurrent episodes of hypoglycemia. Endogenous hyperinsulinism was confirmed and endoscopic ultrasonography identified a pancreatic tumor, which was positive for insulin by immuno-histological staining. After surgical excision of the tumor, no further hypoglycemic attacks occurred. Loss of body weight after removal of the tumor correlated with a dramatic reduction of insulin resistance to such a degree that diet alone proved sufficient for satisfactory glycemic control. PMID- 11388081 TI - [Acceptance of and satisfaction with medical information provided to cancer patients]. AB - BACKGROUND: Medical information to oncologic patients about their disease as well as regularly updated information about the course of their disease and the therapeutic success are essential components of a comprehensive treatment in cancer patients. STUDY GOAL: The quality of the patient-doctor-interaction as well as the hospital preference of oncologic patients were evaluated by a questionnaire at the Oncologic Out-Patient Clinic of the University Hospital of Vienna. METHODS: 350 questionnaires containing 12 questions about medical information, anti-cancer therapy, suggestions for improvement and hospital preference were distributed. The questions were correlated with the patients' demographic and medical data. RESULTS: Out of 350 questionnaires, 234 (67%)--160 (68%) by women and 74 (32%) by men--were returned. 75% of the patients were satisfied with the provided medical information. In contrast, 12% of patients felt incompletely informed about their particular cancer and 19% were unsatisfied with their state of information about the actual status of their disease. Our institution was mostly visited for quality-associated reasons and only in 3% for pragmatic reasons. 58% of the patients had no suggestions for improvement of medical care, although 28% of the patients wanted to spend more time with their doctors, 10% asked for more psychological care and 8% for additional alternative therapeutic modalities. CONCLUSION: The Oncologic Out-Patient Clinic is frequented mainly for quality-associated reasons. Although satisfaction with medical management is very high, there remains space for improvement of information about the underlying disease and its current status. PMID- 11388082 TI - Drug treatment services for ethnic communities in Victoria, Australia: an examination of cultural and institutional barriers. AB - Under-representation of ethnic minorities at drug treatment services represents under-utilisation rather than a lower need. To explore barriers to drug treatment among ethnic communities we undertook a comprehensive review of international and Australian literature to identify problems their members experience upon the discovery of illicit drug use in their community, how drug treatment is addressed and challenges for improved drug treatment outcomes. The concepts and themes derived from the literature were then compared with our research findings from key informant interviews and consultations with non-illicit drug-using spokespersons from eight ethnic communities in Victoria, Australia. Intense shame and loss of face linked to illicit drug use was common in ethnic communities and as a consequence seeking help for drug treatment was fraught with difficulties. Accessing drug treatment services often occurred following a crisis, but a sense of despair and confusion often prevailed owing to a lack of knowledge of available assistance. Even when treatment services were accessed most key informants and ethnic communities viewed them as culturally insensitive, inflexible and with language barriers that obstructed the flow of effective information. Understanding of the ethnic family ethos was of pivotal importance but frequently ignored by treatment services, contributing to the exclusion of ethnic communities from appropriate assistance. Ethnic communities need to be assisted to participate in drug issue discussions and community development strategies in order for their utilisation of drug treatment services to be improved. PMID- 11388083 TI - Racial differences in rates of type 2 diabetes in American women: how much is due to differences in overall adiposity? AB - BACKGROUND: Although the rates of type 2 diabetes attributable to obesity have been documented by numerous studies, little attention has been paid to an important question of, "what fraction of the differences in rates of diabetes between high-risk and low-risk women is due to their differences in prevalence of obesity? This study aimed, therefore, to determine how much of the relative difference in the rates of type 2 diabetes between high-risk non-Hispanic Black and low-risk non-Hispanic White American women can be attributed to differences in overall adiposity. METHODS: Data (n = 1,222) from the Third National Health and Nutrition Examination Survey (1988-94) was used in this investigation. Obesity was defined as body mass index of 30 kg/m2 or more. Estimates of population attributable risks and relative attributable risks were derived using multiple logistic regression models, adjusting for age, smoking and alcohol intake. RESULTS: There were within and between population differences in the impact of obesity on diabetes. The impact of obesity on diabetes as measured by odds ratio and population attributable risk was greater for White women compared to Black women. Obesity was independently associated with a 4-fold (OR = 4.43; 95% CI: 2.65, 7.44) and almost a 2-fold (OR = 1.85; 95% CI: 0.99, 3.47) increased risks of diabetes for White and Black women, respectively. Being Black was associated with an almost 2-fold (OR = 1.86; 95% CI: 1.22, 2.82) increased risk of diabetes relative to White. The population attributable risks of diabetes due to obesity were 49.9% and 28% in Whites and Blacks, respectively. Over one-third (39%) of the difference in rates of diabetes between Black and White women was attributable to differences in the prevalence of overall obesity, adjusting for age, smoking and alcohol consumption. CONCLUSIONS: The result of this study indicating that obesity is a significant factor in explaining the higher prevalence of diabetes among Black women has public health relevance. Since obesity is a preventable and an avoidable risk factor for type 2 diabetes in all populations, the result of this investigation presents a compelling reason to emphasize public health measures to educate women on the need to reduce weight, particularly the high-risk black American women. PMID- 11388084 TI - Supply and segregation of nursing home beds in Chicago communities. AB - OBJECTIVE: Previous research suggests that a shortage of nursing home beds in Latino communities and segregation within facilities in urban settings may contribute to low utilization patterns that both Latino and African American elders exhibit. In order to explore structural barriers to nursing home care for African American and Latino families, this study examines the supply and ethnoracial composition of nursing homes in Chicago communities. DESIGN: With data from the 1990 US Census of Population and Illinois' 1994 Long-Term Care Facility Survey, regression was used to determine if Latino nursing home residents in Chicago follow neighborhood residential patterns in the same way that African American nursing home residents do. Next the availability of nursing home beds by ethnoracial community is examined using analysis of variance. Finally, we present correlations between the racial/ethnic composition of Chicago's facilities, community demographics and facility characteristics that have been associated with quality outcomes. RESULTS: Both African American and Latino nursing home residents follow residential housing patterns, tending to reside in facilities located in their own communities. Latino communities have the fewest beds. However, Latinos appear to be more mobile in their utilization of nursing facilities in other communities than either African Americans or whites and tend to reside in smaller homes with fewer Medicaid recipients. CONCLUSION: Health policy makers must actively address racial and ethnic differences in access to long-term care or risk reinforcing the effects of poverty and segregation. In order to ensure that Latino elders living alone are not going without needed care city leaders must promote a range of culturally sensitive alternatives to nursing home care within Latino communities while promoting geographic mobility for African Americans. PMID- 11388085 TI - Cross-cultural comparisons of health status in Canada using the Health Utilities Index. AB - OBJECTIVE: The purpose of this study was to examine the differences in health status, as measured by the Health Utilities Index (HUI), among seven cultural groups in Canada defined by place of birth and language. DESIGN: The study analysed cross-sectional data from the National Population Health Survey conducted by Statistics Canada in 1994-95. RESULTS: Age-standardized prevalence of dysfunction, defined as HUI < 0.83, varied from 12.7% in English-speaking immigrants to 17.8% in French-speaking Canadians. Considerable differences between the groups were found in the reporting of pain, emotional function, and cognitive function. The variation in HUI scores across the cultural groups could not be explained by differences in socioeconomic status and self-reported chronic conditions. CONCLUSIONS: Although the healthy immigrant effect is probably responsible for some of the variation in health status among cultural groups in Canada, considerable differences exist within the immigrant and Canadian-born populations. Cultural factors may have a substantial effect on the reporting of pain and mental health problems. Further studies are needed to determine the cross-cultural validity of the HUI. PMID- 11388086 TI - High use of sedatives and hypnotics in ethnic minorities in Sweden. AB - OBJECTIVE: To study use of analgesics, and psychotropic drugs in relation to health indicators in four ethnic minorities in Sweden in comparison with Swedish born. DESIGN: Cross-sectional study based on data from the Survey of Living Conditions and Immigrant Survey of Living Conditions in Sweden in 1996. STUDY POPULATION: Random samples of 1890 Swedish residents, in the age range 27-60 years, born in Chile, Poland, Turkey and Iran and 2452 age-matched Swedish-born residents. RESULTS: A two fold higher use of prescribed analgesics and antidepressants and a five to sixfold higher use of hypnotic and sedative drugs was demonstrated in members of ethnic minorities in Sweden in comparison with Swedish-born. In a multivariate analysis the higher use of prescribed analgesics and antidepressants was explained almost entirely by a higher morbidity in the minority study groups. A twofold higher use of sedatives and hypnotics was demonstrated in the minority study populations compared to the Swedish-born sample even after adjustment for extensive indicators of psychiatric and physical health in the multivariate analysis. CONCLUSIONS: The higher use of sedatives and hypnotics in relation to health in the minority samples in the present study indicates a differential treatment of minor psychiatric disorders of members of ethnic minorities in Swedish health services. Further studies that yield more qualitative data regarding the interaction of Swedish physicians with migrant patients are needed to explain these differences and to create a basis for intervention. PMID- 11388087 TI - Ethnic and gender differences in drinking and smoking among London adolescents. AB - OBJECTIVES: The study examines stages of drinking and smoking careers and transitions from initiation to regular use among adolescents, as a function of ethnic status and gender. DESIGN: The data were collected using a confidential, self-completion questionnaire assessing onset and frequency of drinking and smoking. The sample consisted of 1777 adolescents, between the ages of 11 and 14, drawn from eight secondary schools in south-west London. RESULTS: For both smoking and drinking, white children were more likely to have ever smoked tobacco and drunk alcohol, and were also more likely to progress from initiation to regular use than were either black or Asian children. Asian children reported the latest onset and the lowest prevalence rates for both drinking and smoking. Males reported experimenting with both cigarettes and alcohol at an earlier age than females, although a lower proportion of males report regular and lifetime involvement with both alcohol and tobacco. Furthermore, a significantly higher proportion of females who try smoking go on to do so regularly. DISCUSSION: The importance of sociocultural factors in relation to race and gender in predicting onset and escalation of substance use is discussed. The fact that age of onset does not appear to be a significant determinant of transition rate from initiation to regular use is also explored. PMID- 11388088 TI - Cognitive-behavioural therapy for the management of sickle cell disease pain: identification and assessment of costs. AB - OBJECTIVE: The aim of the present study was to investigate the economic validity of using a psychological intervention in the management of sickle cell disease (SCD). Thomas et al. (Br J Health Psychol 1999; 4: 209-29) concluded that cognitive-behaviour therapy (CBT) appears to be immediately effective for the management of SCD pain in terms of reducing psychological distress pain as well as improving coping. METHOD: The costs of management of SCD were evaluated using a societal viewpoint. This approach includes health and social services as well as costs privately borne by informal carers, but it did not include the economic loss due to patients' foregone earnings. Cost profiles were constructed for each patient taking account of cost generating events 12 months before and 12 months after CBT. RESULTS: The hypothesis of the present study, stating that CBT is economically efficient, was confirmed. However, analysis of longitudinal data suggests that CBT is most cost-effective during the first 6 months after the intervention. CONCLUSION: The present findings suggest the need for CBT to be integrated into the normal package of care available for all patients with SCD. The clinical implication is that CBT should be routinely offered to patients on a 6-monthly basis. PMID- 11388089 TI - Noise in a minimal regulatory network: plasmid copy number control. PMID- 11388090 TI - Light at the end of the Ca(2+)-release channel tunnel: structures and mechanisms involved in ion translocation in ryanodine receptor channels. AB - RyR and InsP3R are Ca(2+)-release channels. When induced to open by the appropriate stimulus, these channels allow Ca2+ to leave intracellular storage organelles at an astonishing rate. Investigations of the ion-handling properties of isolated RyR channels have demonstrated that, at least in comparison to voltage-gated channels of surface membranes, these channels display limited powers of discrimination between physiologically relevant cations and this relative lack of selectivity is likely to contribute to the ability of Ca(2+) release channels to maintain high rates of cation translocation without compromising function. A range of ion-handling properties in RyR are consistent with the proposal that this channel functions as a single-ion channel and theoretical considerations indicate that the high rates of ion translocation monitored for RyR would require the pore of such a structure to be short and possess a large capture radius. Measurements of the dimensions of regions of RyR involved in ion conduction and discrimination indicate that this is likely to be the case. In each monomer of RyR/InsP3R, residues making up the last two trans membrane spanning domains and a luminal loop linking these two helices contribute to the formation of the channel pore. The luminal loops of both RyR and InsP3R contain amino acid sequences similar to those known to form the selectivity filter of K+ channels. In addition the luminal loops of both Ca(2+)-release channels contain sequences that are likely to form helices that may be analogous to the pore helix visualised in KcsA. The correlation in structural elements of the luminal loops of RyR/InsP3R and KcsA has prompted us to speculate on the tertiary arrangement for this region of the Ca(2+)-release channels using the established structure of KcsA as a framework. PMID- 11388091 TI - Familial lupus erythematosus. Clinical and immunologic features of 125 multiplex families. AB - Evidence for a genetic susceptibility to systemic lupus erythematosus (SLE) in humans is based on the high concordance rate observed in identical twins and on the relatively high incidence of familial cases. Although recent genetic studies have lead to significant advances in the identification of new susceptibility genes in SLE, no large clinico-pathologic study of familial SLE has been reported to date. In the present study, we describe the main clinical and immunologic features of 125 lupus multiplex families including at least 2 cases of SLE and/or discoid lupus erythematosus (DLE), recruited through a French national survey starting in July 1997. Medical records of all affected members were reviewed by the same investigator, all available family members were interviewed using the same standardized procedure, and blood was drawn for autoantibodies typing. Clinical and immunologic features of 90 probands from multiplex SLE families were compared with those of 100 sporadic SLE patients sharing the same French Caucasian origin. The 125 lupus multiplex families included 282 affected members (2.3 patients per family); of the 125 families, 96 were of French Caucasian origin. One hundred multiplex families included 2 affected relatives, while 25 included 3 or more affected individuals. The relationship between affected members was sibs (45%), parent-offspring (31%), and second-degree (24%). An autosomal dominant mode of inheritance was strongly suggested in 1 extended pedigree with 6 clinically affected members, and a recessive pattern was suspected in 5 other families. No obvious mode of inheritance could be suspected in most of the remainder. Among French Caucasians, sex ratio, mean age at onset, and clinical and biologic SLE-related manifestations were not significantly different in multiplex compared with sporadic SLE cases. The analysis of these 125 multiplex families suggests a genetic heterogeneity that should be considered for ongoing genomic screening. PMID- 11388092 TI - Clinical presentation of left atrial cardiac myxoma. A series of 112 consecutive cases. AB - We analyzed a series of 112 consecutive cases of left atrial myxoma diagnosed in a single French hospital (72 women and 40 men; age range, 5-84 yr) over 40 years, from 1959 to 1998. Symptoms of mitral valve obstruction, the first arm of the classic triad of myxoma presentation, were present in 75 patients (67%), with mostly cardiac failure or malaise. Symptoms of embolism, the second frequent presentation in the classic triad, were observed in 33 cases (29%) with 1 or several locations, essentially cerebral emboli with stroke. Males are statistically at greater risk than females of developing embolic complications. The third arm of the classic triad consists of constitutional symptoms (34%) with fever, weight loss, or symptoms resembling connective tissue disease, due to cytokine (interleukin-6) secretion. Younger and male patients have more neurologic symptoms, and female patients have more systemic symptoms. Seventy-two patients (64%) had cardiac auscultation abnormalities, essentially pseudo-mitral valve disease (53.5%) and more rarely the suggestive tumor plop (15%). The most frequent electrocardiographic sign was left atrial hypertrophy (35%), whereas arrhythmias were uncommon. The greater number of myxoma patients (98) diagnosed preoperatively after 1977 reflects the introduction of echocardiography as a noninvasive diagnostic procedure. However, there was no significant reduction in the average time from onset of symptoms to operation between patients seen in the periods before and after 1977. The tumor diameter ranged from 1 to 15 cm with a weight of between 15 and 180 g (mean, 37 g). The myxoma surface was friable or villous in 35% of the cases, and smooth in the other 65% cases. Myxomas in patients presenting with embolism have a friable surface; those in patients with cardiac symptoms, pseudo-mitral auscultation signs, tumor plop, and electrocardiogram or radiologic signs of left atrium hypertrophy and dilatation are significantly the larger tumors. The long-term prognosis is excellent, and only 4 deaths occurred among our 112 cases over a median follow-up of 3 years. The recurrence rate is low (5%), but long-term follow-up and serial echocardiography are advisable especially for young patients. PMID- 11388093 TI - Dermatologic manifestations of relapsing polychondritis. A study of 200 cases at a single center. AB - Dermatologic manifestations of relapsing polychondritis (RP) have been relatively poorly studied compared to other manifestations. In this study we describe dermatologic manifestations in a large series of patients with RP and the corresponding pathologic findings. In this retrospective, single-center review of 200 patients diagnosed with RP according to Michet's criteria, we analyzed separately those suffering from associated diseases with potential dermatologic involvement or chronic dermatitis. Skin or mucosal biopsies taken from 59 patients were examined without knowledge of the clinical data. Among the 200 patients with RP, 73 had chronic dermatitis or associated diseases with potential dermatologic involvement, especially hematologic disorders (n = 24) and connective tissue diseases (n = 22). Among the other 127 patients, 45 (35.4%) had dermatologic manifestations: aphthosis (n = 21; oral in 14 and complex in 7), nodules on the limbs (n = 19), purpura (n = 13), papules (n = 10), sterile pustules (n = 9), superficial phlebitis (n = 8), livedo reticularis (n = 7), ulcerations on the limbs (n = 6), and distal necrosis (n = 4). Dermatologic manifestations were the presenting feature of RP in 15 cases (12%), and appeared concomitantly (n = 23) or not (n = 22) with attacks of chondritis. Histologic findings included vasculitis (n = 19, leukocytoclastic in 17 and lymphocytic in 2), neutrophil infiltrates (n = 6), thrombosis of skin vessels (n = 4), septal panniculitis (n = 3), and minor changes (n = 2). Patients with and without dermatologic manifestations did not differ with regard to male/female ratio; age at RP onset; frequency of auricular, nasal, or tracheobronchial chondritis; or frequency of rheumatologic, ocular, audiovestibular, renal, arterial, or venous involvement. The frequency of dermatologic manifestations (91% versus 35.4%; p < 0.0001), sex ratio (18 male/4 female versus 44 male/83 female, p < 0.0001), and age at first chondritis (63.3 +/- 14 yr versus 41.4 +/- 17 yr; p < 0.0002) were significantly higher in the 22 patients with myelodysplastic syndrome than in the 127 patients without any associated disease. In conclusion, although dermatologic manifestations occur frequently in patients with RP, especially in association with myelodysplasia, they are nonspecific and sometimes resemble those observed in Behcet disease or inflammatory bowel diseases. Their presence in the elderly warrants repeated blood cell counts to detect a smouldering myelodysplasia. PMID- 11388094 TI - Clinical and histopathologic features and immunologic variables in patients with severe chilblains. A study of the relationship to lupus erythematosus. AB - We investigated 33 patients affected with chilblain lesions following a persisting course of more than 1 month. We focused on the incidence of an underlying connective tissue disease, mostly lupus erythematosus (LE), and we analyzed features of idiopathic chilblains compared with those of chilblain lesions associated with connective tissue disease. We also carried out a prospective follow-up of patients. Eleven patients included in the study were free of any clinical and/or laboratory abnormality suggestive of connective tissue disease, while 22 of 33 patients showed 1 or several abnormalities raising suspicion for connective tissue disease, and among them 8 had a diagnosis of systemic lupus erythematosus (SLE) established at initial evaluation based on the American College of Rheumatology revised criteria. The comparative analysis of patients with idiopathic chilblains and patients with chilblains associated with LE showed that female sex and persistence of lesions beyond cold seasons were significantly associated with chilblain LE. Histopathologic studies of chilblain lesions did not reveal features typical of LE in any case, but revealed a higher incidence of a deep perisudoral infiltrate in idiopathic chilblains. In patients showing signs of connective tissue disease, positive cutaneous immunofluorescence was correlated with the presence of circulating antinuclear antibodies. Two patients had an ascertained diagnosis of SLE with severe manifestations during prospective follow-up, requiring treatment with oral steroids in both cases. Chilblains following a chronic course may reveal connective tissue disease, and patients affected with chilblains associated with autoimmune abnormalities may develop severe SLE. Accordingly, long-term follow-up of these patients is warranted. PMID- 11388096 TI - The NASPE/BPEG Codes: use, misuse, and evolution. PMID- 11388095 TI - Gastric secretion in Zollinger-Ellison syndrome. Correlation with clinical expression, tumor extent and role in diagnosis--a prospective NIH study of 235 patients and a review of 984 cases in the literature. AB - We prospectively studied 235 patients with Zollinger-Ellison syndrome (ZES) (205 without and 30 with prior acid-reducing surgery) and compared the results with 984 patients from 182 reports in the literature. The aims of the study were to evaluate the sensitivity of proposed acid secretory criteria for the diagnosis of ZES, propose new criteria, evaluate the variability and methodology of gastric secretory testing, and correlate the symptoms and signs of ZES, tumor extent, and primary tumor size and location with the degree of gastric acid hypersecretion. Multiple endocrine neoplasia-type 1 (MEN1) occurred in 22% of patients. The mean basal acid output (BAO) in patients without and with prior acid-reducing surgery was 41.2 +/- 1.7 mEq/hr (range, 1.6-118.3 mEq/hr) and 27.6 +/- 3.5 mEq/hr (range 5.9-102.9 mEq/hr), respectively. In patients with MEN1, those with female gender, Hispanic, or Asian race had lower BAOs. Diarrhea, esophageal stricture, and pyloric scarring were associated with a higher BAO. Neither other symptoms nor the tumor extent, primary tumor location, or size correlated with the magnitude of acid hypersecretion. ZES diagnosis was delayed a mean of 5.5 +/- 0.4 yr. Patients who were misdiagnosed as having either Crohn or celiac disease had higher BAOs. The sensitivities from our study and the literature review of the proposed BAO criteria for the diagnosis of ZES in patients without previous gastric acid-reducing surgery were 91% and 90% for BAO > or = 15 mEq/hr, 86% and 82% for BAO > or = 18 mEq/hr, 69% and 67% for BAO > 25 mEq/hr, and < 60% for BAO > 31 mEq/hr, respectively. The specificities of all the proposed BAO criteria were high. Both the criterion of BAO > or = 15 mEq/hr and BAO > or = 18 mEq/hr had good specificities and equal sensitivity. With prior acid-reducing surgery, the sensitivities in our study and from the literature review were 100% and 81% for BAO > or = 5 mEq/hr, 73% and 45% for BAO > 14.4 mEq/hr, and 37% and 31% for BAO > 19.2 mEq/hr, respectively. The reported mean specificity for the criterion of BAO > or = 5 mEq/hr was 85%, while it was 100% for the other 2 criteria. The maximal acid output (MAO) criterion of > 70 mEq/hr had sensitivities in the present National Institutes of Health (NIH) study and the literature review of 39% and 31%, respectively, and the criterion of MAO > 100 mEq/hr had a sensitivity of < 15% in patients with no prior acid-reducing surgery. The proposed criterion of BAO/MAO ratio > 0.6 had a low sensitivity. The proposed criterion of the ratio of basal and maximal acid H+ concentration (BAC/MAC ratio) > or = 0.6 had an excellent sensitivity-- > or = 89% in patients with or without previous acid-reducing surgery. The reported specificity for both the BAO/MAO criterion and the BAC/MAC criterion were similar, but BAC/MAC had a better sensitivity. Combination criteria of BAO generally did not improve sensitivity. The criterion of pH < or = 1 was met by only 27% of patients, and pH < or = 0.96 by 21% of patients with previous acid-reducing surgery. For patients with MEN1 with no prior acid-reducing surgery, the sensitivities were lower compared with patients with the sporadic form of ZES. The mean gastric volume in patients without prior acid-reducing surgery was 314 +/- 10 mL/hr and 247 +/- 25 mL/hr in patients with prior acid-reducing surgery. A basal volume criteria of > 160 mL/hr in patients without prior acid-reducing surgery occurred in > 86% of patients, and > 140 mL/hr in 87% of patients with prior acid-reducing surgery; these, thus, are neglected findings that have good sensitivities. Our analysis shows criteria based on MAO, pH, and BAO/MAO ratio do not have high sensitivities and thus are not useful. In patients without prior acid-reducing surgery, the criteria of BAO > or = 15 mEq/hr, BAC/MAC ratio > or = 0.6, and basal gastric volume > 160 mL/hr are useful for the diagnosis of ZES and have good specificities. In patients with prior acid-reducing surgery, the criteria of BAO > or = 5 mEq/hr, BAC/MAC ratio > or = 0.6, and basal gastric volume > 140 mL/hr have high sensitivities. In patients with sporadic ZES without acid-reducing surgery, the criterion of BAO > or = 18 mEq/hr is recommended as it has a similar sensitivity but higher specificity than the criterion of BAO > or = 15 mEq/hr. Only 1 patient in either data set (NIH or the literature) with or without previous acid-reducing surgery had a basal gastric pH > 2, therefore this finding essentially excludes the diagnosis of ZES. Gastric secretory measurements for 30 minutes, but not 15 minutes, give results comparable to those for a full hour. On the basis of these results, a number of gastric secretory criteria are proposed, including some for the first time, and alterations in methodology are proposed that should prove useful in the diagnosis of ZES. PMID- 11388097 TI - Prolonged QRS duration increases QT dispersion but does not relate to arrhythmias in survivors of acute myocardial infarction. AB - QT dispersion has been suggested and disputed as a risk marker for ventricular arrhythmias after myocardial infarction. Delayed ventricular activation after myocardial infarction may affect arrhythmic risk and QT intervals. This study determined if delayed activation as assessed by (1) QRS duration in the 12-lead ECG and by (2) late potentials in the signal-averaged ECG affects QT dispersion and its ability to assess arrhythmic risk after myocardial infarction. QT duration, JT duration, QT dispersion, and JT dispersion were compared to QRS duration in the 12-lead ECG and to late potentials in the signal-averaged ECG recorded in 724 patients 2-3 weeks after myocardial infarction. Prolonged QRS duration (> 110 ms) and high QRS dispersion increased QT and JT dispersion by 12% 15% (P < 0.05). Presence of late potentials, in contrast, did not change QT dispersion. Only the presence of late potentials (n = 113) was related to arrhythmic events during 6-month follow-up. QT dispersion, JT dispersion, QRS duration, and QRS dispersion were equal in patients with (n = 29) and without arrhythmic events (QT disp 80 +/- 7 vs 78 +/- 1 ms, JT disp 80 +/- 6 vs 79 +/- 2 ms, mean +/- SEM, P > 0.2). In conclusion, prolonged QRS duration increases QT dispersion irrespective of arrhythmic events in survivors of myocardial infarction. Presence of late potentials, in contrast, relates to arrhythmic events but does not affect QT dispersion. Therefore, QT dispersion may not be an adequate parameter to assess arrhythmic risk in survivors of myocardial infarction. PMID- 11388098 TI - Clinical significance of the atrial fibrillation threshold in patients with paroxysmal atrial fibrillation. AB - AF threshold and the other electrophysiological parameters were measured to quantify atrial vulnerability in patients with paroxysmal atrial fibrillation (PAF, n = 47), and those without AF (non-PAF, n = 25). Stimulations were delivered at the right atrial appendage with a basic cycle length of 500 ms. The PAF group had a significantly larger percentage of maximum atrial fragmentation (%MAF, non-PAF: mean +/- SD = 149 +/- 19%, PAF: 166 +/- 26%, P = 0.009), fragmented atrial activity zone (FAZ, non-PAF: median 0 ms, interquartile range 0 20 ms, PAF: 20 ms, 10-40 ms, P = 0.008). Atrial fibrillation threshold (AF threshold, non-PAF: median 11 mA, interquartile range 6-21 mA, PAF: 5 mA, 3-6 mA, P < 0.001) was smaller in the PAF group than in the non-PAF group. Sensitivity, specificity, and positive predictive value of electrophysiological parameters were as follows, respectively: %MAF (cut off at 150%, 78%, 52%, 76%), FAZ (cut off at 20 ms, 47%, 84%, 85%), AF threshold (cut off at 10 mA, 94%, 60%, 81%). There were no statistically significant differences between the non-PAF and PAF groups in the other parameters (effective refractory period, interatrial conduction time, maximum conduction delay, conduction delay zone, repetitive atrial firing zone, wavelength index), that were not specific for PAF. In conclusion, the AF threshold could be a useful indicator to evaluate atrial vulnerability in patients with AF. PMID- 11388099 TI - Clinical relevance of loss of atrial sensing in patients with single lead VDD pacemakers. AB - During single lead VDD pacing, loss of atrial sensing is reported to be 2%-11% of patients in the literature. The impact on quality-of-life and exercise duration has never been established. This study tried to assess the clinical relevance of loss of atrial sensing in patients with single lead VDD pacemakers. Twenty-one patients with total AV block were studied 3 months after single lead VDD pacemaker implantation. During a 6-minute walk test, atrial undersensing was verified and adjusted to achieve undersensing in < 5% (group 1), 5%-10% (group 2), and > 10% (group 3). Patients were allocated to each group in a randomized double blind crossover design. At the end of each 2-month study period all patients underwent symptom limited treadmill exercise testing. Subjective assessment of exercise difficulty was made using the Borg score, and quality-of life assessment was performed using the Health Status Questionnaire. Exercise duration significantly decreased in group 3 (228 +/- 50 s) as compared to group 1 (257 +/- 42 s) and group 2 (250 +/- 46 sec) with an increase in the Borg score. Quality-of-life was decreased for three subscales in group 3 as compared to group 1 and group 2. In conclusion, atrial undersensing of > 10% in patients with single lead VDD pacing was associated with a decrease in exercise duration and increase in the subjective severity score, in addition to a decrease in quality of-life. Atrial undersensing of < 10% did not effect exercise test results or quality-of-life. Since all studies reported intact atrioventricular synchrony in > or = 90% of patients, loss of atrial sensing is of limited clinical importance in these patients. PMID- 11388100 TI - Longevity of dual chamber pacemakers: device and patient related determinants. AB - In 382 patients with three different dual chamber pulse generators, the median time interval to battery depletion was 98.3 months. Cox regression analysis revealed the following variables as significant predictors of battery longevity: programmed pacing rate, energy of the stimulation output, mode of stimulation (i.e., proportion of paced cycles in one or two chambers), battery capacity, and internal sensing current of the pacemaker. Although 27% of all patients died before the service life of the pacemaker was over and despite a rate of premature reoperations of 8.6%, the majority of pacemaker patients (55%) fully used the expected battery life span of the pulse generator. Patients who died before the pacemaker had reached its end of service were significantly older at implantation than patients who survived until pacemaker replacement. The vast majority (92%) of patients received another dual chamber pulse generator when replacement was required. These data underline the need for long-lasting dual chamber devices. PMID- 11388101 TI - Comparison of the rate dependent effects of dofetilide and ibutilide in the newborn heart. AB - This study compared the rate dependent changes in atrial and ventricular monophasic action potential duration in the newborn canine heart in response to two Class III antiarrhythmic agents: dofetilide, a pure Ikr blocker, and ibutilide, a Na+ channel opener. Newborn dogs were anesthetized with pentobarbital, vagotomized, and given propranolol to eliminate autonomic responses. A 4 Fr electrical catheter was placed in the right atrium for pacing. Monophasic action potential durations (APDs) at 90% repolarization (APD90) were recorded from the epicardial surface of the left ventricle and atrium with Ag AgCl2 suction electrodes. APD90 was measured as cardiac cycle length was shortened by pacing, in the control condition and following two doses of dofetilide (n = 8) or ibutilide (n = 9). Slopes of the APD90 versus decreasing paced cycle length (PCL) relationships were then compared. Large dose dependent increases in atrial and ventricular APD90 were observed after dofetilide and ibutilide. In the neonatal atrium, there were no changes in the APD90 versus PCL relationship with either drug, indicating no rate dependency of drug effect. In contrast, in the ventricle, a steeper APD90 versus PCL slope was noted after dofetilide and ibutilide, indicating a significant loss of drug effect at faster heart rates (i.e., reverse rate dependency). In spite of probable different cellular mechanisms of action, the rate dependent characteristics of dofetilide and ibutilide are identical in the neonatal heart. There is no evidence of (reverse) rate dependency in the atrium, predicting that both agents would be effective at rapid atrial tachycardia rates. For both, however, marked reverse rate dependency is observed in the neonatal ventricle. PMID- 11388102 TI - Mechanisms of antiarrhythmic drug action on termination of atrial flutter. AB - Atrial flutter is an important arrhythmia in clinical practice. Although the reentrant circuit of human typical atrial flutter is well characterized, the action of antiarrhythmic drugs on this tachycardia is less understood. Based on the recent clinical trials, pure Class III drugs like ibutilide or dofetilide are more effective in acute termination of human atrial flutter than Class I drugs like procainamide or flecainide. The mechanisms of drug induced termination of atrial flutter include refractory block due to cycle length oscillation, fixed block due to a reduced safety factor for conduction, or a collision of opposing wavefronts due to loss of the lateral boundaries or return reexcitation. Because ventricular proarrhythmia is a major concern with ibutilide or dofetilide therapy, development of new drugs with more specific target profiles is a future direction for treatment of human atrial flutter. PMID- 11388103 TI - Inappropriate detection of supraventricular arrhythmias by implantable dual chamber defibrillators: a comparison of four different algorithms. AB - Inappropriate therapy of supraventricular tachyarrhythmias by an ICD is still a common problem. Dual chamber (DDD) ICDs provide additional atrial sensing and should result in higher specificity for detection of supraventricular tachyarrhythmias. However, a direct comparison of different dual chamber algorithms has not been reported. The detection algorithms of four different DDD ICDs were tested: Phylax AV, Defender IV, Ventak AV III DR, and Gem DR 7271. Based on arrhythmias recorded from patients undergoing invasive electrophysiological studies and in many cases of catheter ablation at our institution, a library consisting of 71 supraventricular and 15 ventricular tachyarrhythmias was created. The library consists of episodes of atrial fibrillation, atrial flutter with different AV conduction, typical and atypical AV nodal reentrant tachycardia, AV reentrant tachycardia, sinus tachycardia, and ventricular tachycardia with and without ventriculoatrial conduction. Atrial fibrillation was appropriately classified by all four algorithms. However, the specificity for detection of other supraventricular tachyarrhythmias achieved by the Biotronik (12%) and the Guidant (11%) devices was significantly lower compared to the specificity of the ELA (28%) and the Medtronic DDD ICD (20%). This is due to the fact that the Biotronik and the Guidant algorithm classified all supraventricular tachyarrhythmias resulting in a stable ventricular rate as ventricular tachycardia, whereas the ELA and Medtronic algorithms performed a more detailed analysis by assessment of PR association, atrial onset, or timing of the atrial event relative to the ventricular event, respectively. Atrial fibrillation, the most common supraventricular tachyarrhythmia in patients with ICD, was detected by all devices. PMID- 11388104 TI - Survey of cardiac pacing and implanted defibrillator practice patterns in the United States in 1997. AB - A survey of implanters of permanent cardiac pacemakers and ICDs in the United States during 1997 was conducted to identify present and changing patterns in indications for pacing, implantation techniques, pacing-mode selection, follow up, and opinions regarding pacing and ICD related issues. This report is an update from 1993 of surveys performed every 4 years for the International Cardiac Pacing and Electrophysiology Society (ICPES). Questionnaires were sent to implanting physicians who were members of the North American Society of Pacing and Electrophysiology (NASPE), and who might, therefore, be expected to be more conversant than others with the state of the art. Four major manufacturers also provided estimates of the numbers of pacemakers and ICDs implanted in the United States from 1994 through 1997. In 1997, approximately 182,000 new rhythm management devices, including 153,000 primary pacing systems and 29,000 ICDs, were implanted, an increase of 24% for pacemakers and 90% for ICDs since 1994. In 1997, pacemaker implantations were performed by about 8,600 physicians working in 3,300 hospitals and 1,000 independent "surgi-centers." From 1994 to 1997, sales in the United States of dual chamber pacemakers rose from 58% to 69% of the total, and adaptive rate systems from 74% to 90%. ICD sales increased by about 29% per year from 18,700 to 35,000 units. This study disclosed significant differences among implanter subcategories and between present and earlier practices, and it provided useful insights into trends in pacemaker and defibrillator practice. Future surveys would be facilitated if a standardized implant registry like that used in Europe were established in the United States. PMID- 11388105 TI - The World Survey of Cardiac Pacing and Cardioverter Defibrillators: calendar year 1997--Asian Pacific, Middle East, South America, and Canada. AB - A comprehensive cardiac pacing and ICD survey of the Asian Pacific region, Middle East, South America, and Canada was undertaken for calendar year 1997. Thirteen countries in the Asian Pacific region contributed to the survey. There were also two countries from the Middle East, three from South America, and Canada. Accurate numbers of new implants and replacements were obtained for cardiac pacemakers. In the Asian Pacific region, Japan with almost 20,000 implants, had the largest number of pacemaker implants, but Australia had the highest implant rate with 345 new implants per million population. However, Canada and Uruguay had higher new implant rates per million. The major indication for implantation of a cardiac pacemaker was high degree atrioventricular block. There was a high incidence of single chamber ventricular pacing in the developing countries of Asia. Only a few countries used substantial numbers of single lead VDD and AAIR pacing systems. Pacing leads were predominantly transvenous, bipolar, passive fixation, and steroid eluting. Active-fixation leads were also used in the atrium. With ICD use, Australia had the highest new implant rate per million population. The major indications were sudden death and ventricular arrhythmias. A group of enthusiastic survey coordinators has now been established. It is hoped to continue physician and country recruitment, particularly in the Middle East and South American regions to establish a fully global experience of cardiac pacing and ICD use. PMID- 11388106 TI - The World Survey of Cardiac Pacing and Implantable Cardioverter Defibrillators: calendar year 1997--Europe. AB - The registry of the European Working Group on Cardiac Pacing (EWGCP) is based on the European Pacemaker Identification Card originally designed in July 1978. National registration centers collect the local data and send aggregated annual data to the EWGCP. For 1997, data were obtained from 2,887 hospitals in 20 European countries representing a population of 568 million. Across all participating countries, the median value for all implanted pacemakers was 378 per million population. For initial pacemaker implants, the median value was 290 per million population. Single chamber atrial pacing was important in Denmark, the Netherlands, Poland, Slovak Republic, Spain, and Sweden for the treatment of sick sinus syndrome. Dual chamber pacing accounted for < 50% of initial implants in only 5 of 14 countries for atrioventricular block, and in only 3 of 15 countries for sick sinus syndrome. In 7 of 15 countries, unipolar ventricular leads were used in > or = 50% of cases. In 6 of 14 countries, there was > 15% use of unipolar atrial leads. Nine of 13 countries frequently used atrial active fixation leads. For the 1997 survey, ICD data were obtained from 16 countries. The total number of ICDs per million population was a median value of 14. Initial ICD implants per million population was 11. Only 3 of 16 countries implanted a total of 30 or more ICDs per million population. Pacing and ICD practices were dependent on the availability of medical and technical resources and influenced by economic constraints inherent in health care administration and insurance coverage patterns. PMID- 11388107 TI - The World Survey of Cardiac Pacing and Cardioverter Defibrillators: calendar year 1997. PMID- 11388108 TI - Sudden death in heart failure associated with reduced left ventricular function: substrates, mechanisms, and evidence-based management, Part I. PMID- 11388109 TI - SVT or VT? Diagnostic dilemma in a patient with a dual chamber implantable cardioverter defibrillator. PMID- 11388110 TI - The dignity of life and death and the power of music. PMID- 11388111 TI - Transient exit block of a DDD pacemaker with unipolar leads in subcutaneous emphysema following pneumothorax. AB - This case report describes a transient pacemaker exit block due to subcutaneous emphysema following pneumothorax. Pneumothorax after pacemaker implantation is rare, but development of subcutaneous emphysema under such circumstances is even more uncommon. Exit block develops only with the use of unipolar leads; with implantation of bipolar leads, this complication cannot occur. PMID- 11388112 TI - Levofloxacin induced polymorphic ventricular tachycardia with normal QT interval. AB - Polymorphic ventricular tachycardia (PVT) is a form of ventricular tachycardia characterized by QRS complexes that seem to change direction during the tachycardia. If associated with a prolonged QT interval, it is called torsades de pointes. In the absence of a congenital long QT syndrome, torsades is seen with certain drugs such as antiarrhythmic agents (Class IA, IC, III), psychotropic medications, antidepressants, antihistamines, and electrolyte disturbances. We report the first case of polymorphic ventricular tachycardia with normal QT interval associated with the oral use of levofloxacin in the absence of other etiologies known to cause these arrhythmias. PMID- 11388113 TI - Adenosine- and verapamil-sensitive ventricular tachycardia in the newborn. AB - Two neonates presented with sustained, monomorphic VT. Transesophageal electrophysiological studies demonstrated that the VTs were initiated with burst atrial pacing in one and noninducible in the other, and both terminated with burst atrial pacing and with adenosine. Oral verapamil suppressed the VTs in both. Following discontinuation of verapamil at 1 year of age, both children remain free of tachycardia recurrence at 3 and 4 years of age. These cases suggest that cAMP-mediated triggered activity may be responsible for some VTs in infancy. PMID- 11388114 TI - Relation between the rapid focal activation in the pulmonary vein and the maintenance of paroxysmal atrial fibrillation. AB - This case report describes a patient with drug refractory paroxysmal atrial fibrillation (PAF). Rapid focal activations with multiple sharp spikes were continuously identified inside the left superior pulmonary vein (PV) during sustained AF. Among seven episodes of AF, cessation of rapid focal activations coincided with the conversion of AF to flutter (n = 4) or immediate AF termination (n = 3). Guided by sharp spikes in the PV, abrupt termination of AF occurred during radiofrequency energy application. Conclusively, rapid focal activations inside the PV are critical in AF maintenance. Cessation of these rapid focal activations underlies the mechanism by which AF converts to flutter. PMID- 11388115 TI - Paradoxical induction of cross-talk in a dual chamber pacemaker by increasing the ventricular output. AB - Cross-talk was induced in a unipolar DDD pacemaker by increasing the output of the ventricular channel. Oversensing by the ventricular channel was probably due to the detection of a relatively large summation signal consisting of the afterpotential and the superimposed activity from the atrial stimulus. PMID- 11388116 TI - Hyperkalemia diagnosed by implantable cardioverter defibrillator T wave sensing. AB - A patient with congestive heart failure and an ICD had undergone atrioventricular nodal ablation and optimization of heart failure medical therapy. Intracardiac T wave sensing by the ICD drew attention to the new development of asymptomatic hyperkalemia. Surface ECG features of hyperkalemia were not readily identified due to pacemaker dependence. PMID- 11388117 TI - Recurrent pacemaker lead induced axillary subclavian vein thrombosis. AB - This report describes a patient with two recurrences of axillary subclavian vein thrombosis more than 1 year after implantation of a permanent transvenous pacemaker. Both recurrences were successfully treated with local thrombolysis. PMID- 11388118 TI - Spontaneous reattachment of a dislodged atrial lead. AB - Acute lead dislodgment is a complication that should be managed immediately. Spontaneous reattachment of a dislodged atrial pacing lead has never been reported before. This case report describes a lucky patient whose atrial lead spontaneously gained a new functioning position after dislodgment from its initial implantation site. PMID- 11388119 TI - Neuropsychological function in patients with chronic fatigue syndrome, multiple sclerosis, and depression. AB - Patients with chronic fatigue syndrome (CFS), multiple sclerosis (MS), and major depression were compared with controls and with each other on a neuropsychological battery that included standard neuropsychological tests and a computerized set of tasks that spanned the same areas of ability. A total of 101 participants were examined, including 29 participants with CFS, 24 with MS, 23 with major depressive disorder, and 25 healthy controls. There were significant differences among the groups in 3 out of 5 cognitive domains: memory, language, and spatial ability. Assessment of psychiatric symptoms indicated that all 3 patient groups had a higher prevalence of depression than the controls. A total measure of psychiatric symptomatology also differentiated the patients from the controls. After covarying the cognitive test scores by a measure of depression, the patient groups continued to differ from controls primarily in the area of memory. The findings support the view that the cognitive deficits found in CFS cannot be attributed solely to the presence of depressive symptomatology in the patients. PMID- 11388120 TI - Quantitative assessment of cerebral ventricular volumes in chronic fatigue syndrome. AB - Previous qualitative volumetric assessment of lateral ventricular enlargement in chronic fatigue syndrome (CFS) has provided evidence for subtle structural changes in the brains of some individuals with CFS. The aim of this pilot study was to determine whether a more sensitive quantitative assessment of the lateral ventricular system would support the previous qualitative findings. In this study, we compared the total lateral ventricular volume, as well as the right and left hemisphere subcomponents in 28 participants with CFS and 15 controls. Ventricular volumes in the CFS group were larger than in control groups, a difference that approached statistical significance. Group differences in ventricular asymmetry were not observed. The results of this study provide further evidence of subtle pathophysiological changes in the brains of participants with CFS. PMID- 11388121 TI - Cognitive compromise following exercise in monozygotic twins discordant for chronic fatigue syndrome: fact or artifact? AB - This study examined the effects of exhaustive exercise on cognitive functioning among 21 monozygotic twin pairs discordant for chronic fatigue syndrome (CFS). The co-twin control design adjusts for genetic and family environmental factors not generally accounted for in more traditional research designs of neuropsychological function. Participants pedaled a cycle ergometer to exhaustion; maximum oxygen output capacity (VO2max) as well as perceived exertion were recorded. Neuropsychological tests of brief attention and concentration, speed of visual motor information processing, verbal learning and recognition memory, and word and category fluency were administered with alternate forms to participants pre- and postexercise. The preexercise neuropsychological test performance of CFS twins tended to be slightly below that of the healthy twin controls on all measures. However, twins with CFS did not demonstrate differential decrements in neuropsychological functioning after exercise relative to their healthy co-twins. Because exercise does not appear to diminish cognitive function, rehabilitative treatment approaches incorporating exercise are not contraindicated in CFS. PMID- 11388122 TI - Divided attention deficits in patients with chronic fatigue syndrome. AB - Chronic fatigue syndrome (CFS) patients and controls were compared on a variety of mood state, personality, and neuropsychological measures, including memory, word finding, and attentional tasks that required participants to focus, sustain, or divide their attention, or to perform a combination of these functions. CFS patients demonstrated a selective deficit on 3 measures of divided attention. Their performance on the other neuropsychological tests of intelligence, fluency, and memory was no different than that of normal controls despite their reports of generally diminished cognitive capacity. There was an inverse relation between CFS patient fatigue severity and performance on 1 of the divided attention measures. Given these findings, it is probable that CFS patients will report more cognitive difficulties in real-life situations that cause them to divide their effort or rapidly reallocate cognitive resources between 2 response channels (vision and audition). PMID- 11388123 TI - Longitudinal assessment of neuropsychological functioning, psychiatric status, functional disability and employment status in chronic fatigue syndrome. AB - The longitudinal course of subjective and objective neuropsychological functioning, psychological functioning, disability level, and employment status in chronic fatigue syndrome (CFS) was examined. The relations among several key outcomes at follow-up, as well as the baseline characteristics that predict change (e.g., improvement), were also evaluated. The study sample consisted of 35 individuals who met the 1988 and 1994 CFS case definition criteria of the Centers for Disease Control (CDC) at intake. Participants were evaluated a mean of 41.9 (SEM = 1.7) months following their initial visit (range = 24-63 months). Results indicated that objective and subjective attention abilities, mood, level of fatigue, and disability improve over time in individuals with CFS. Moreover, improvements in these areas were found to be interrelated at follow-up. Finally, psychiatric status, age, and between-test duration were significant predictors of outcome. Overall, the prognosis for CFS appears to be poor, as the majority of participants remained functionally impaired over time and were unemployed at follow-up, despite the noted improvements. PMID- 11388124 TI - Cytokine and other immunologic markers in chronic fatigue syndrome and their relation to neuropsychological factors. AB - The literature is reviewed and data are presented that relate to a model we have developed to account for the perpetuation of the perplexing disorder currently termed chronic fatigue syndrome (CFS). In patients with CFS there is chronic lymphocyte overactivation with cytokine abnormalities that include perturbations in plasma levels of proinflammatory cytokines and decrease in the ratio of Type 1 to Type 2 cytokines produced by lymphocytes in vitro following mitogen stimulation. The initiation of the syndrome is frequently sudden and often follows an acute viral illness. Our model for the subsequent chronicity of this disorder holds that the interaction of psychological factors (distress associated with either CFS-related symptoms or other stressful life events) and the immunologic dysfunction contribute to (a) CFS-related physical symptoms (e.g., perception of fatigue and cognitive difficulties, fever, muscle and joint pain) and increases in illness burden and (b) impaired immune surveillance associated with cytotoxic lymphocytes with resulting activation of latent herpes viruses. PMID- 11388125 TI - Patients' knowledge of health plan coverage and satisfaction with care. AB - OBJECTIVE: To test the hypothesis that patients' satisfaction with their healthcare is related to their knowledge of their managed care plan. STUDY DESIGN: A written survey was sent to beneficiaries of the military health system. PATIENTS AND METHODS: Respondents were active or retired military personnel and their nonmilitary, immediate family members enrolled in either TRICARE Prime, a voluntary, gatekeeper-based managed care program (enrollees), or other military managed care programs (nonenrollees). Responses to 5 questions that measured patients' understanding of their program served as independent variables; satisfaction with care was the dependent variable. Responses were stratified according to whether care was received in a military or a civilian healthcare organization. Analysis of variance (ANOVA) and regression analysis were used to determine the association between the variables. RESULTS: Enrollees differed from nonenrollees in certain sociodemographic characteristics, with nonenrollees being older (eta 2 = 0.035; P < .01), closer to retirement (eta 2 = 0.051; P < .01), and tending to receive healthcare in military healthcare organizations (HCO) (eta 2 = 0.009; P < .01). After controlling for sociodemographic characteristics, patients' understanding of their coverage served as a poor predictor (military HCO [R2 = 0.003; P < .01]; civilian HCO [R2 = 0.025; P < .01]) of satisfaction with their care. CONCLUSIONS: Patients' understanding of their coverage appears to be a statistically significant, but rather small, contributor to satisfaction with their care. The study suggests that health plan understanding is a poor predictor of patient satisfaction. PMID- 11388126 TI - Development and validation of the pharmaceutical care satisfaction questionnaire. AB - The development and validation of a survey instrument to assess consumer satisfaction with pharmacy services is discussed. The Pharmaceutical Care Satisfaction Questionnaire (PCSQ) is a 30-item instrument administered by someone other than the pharmacist that uses a Likert scale to score respondents' answers. The PCSQ is written approximately at a seventh grade reading level. Following initial development, the PCSQ was administered to 360 consumers in ambulatory pharmacies and to 311 patients in a multicenter hyperlipidemia outcomes study. The Cronbach coefficient alpha was .94 for the consumer data, with a 64.8% variance accounted for by the 4-factor solution. A coefficient alpha of .84 was found on all 30 items in the hyperlipidemia study, with a variance of 63.78% in control patients and 60.16% in treatment patients. The PCSQ is easy to administer and score, with minimal cost. Unlike other satisfaction surveys, the PCSQ contains patient evaluations regarding outcomes of care. A primary limitation of the PCSQ is that it is a newly developed instrument that needs to be used in more studies to strengthen its validity. PMID- 11388127 TI - Health insurance, primary care, and preventable hospitalization of children in a large state. AB - OBJECTIVES: To analyze variations in the admission rate to hospitals of children with ambulatory care-sensitive (ACS) conditions and to test the relationship of ACS admission rates to insurance coverage, health maintenance organization (HMO) enrollment, availability of primary care, severity of illness, distance to hospital, and a number of other factors. STUDY DESIGN: Hypotheses were derived from basic considerations about demand and use of primary care and preventive services and then were tested with a weighted linear regression model of the ACS admission rate for children residing in each county. PATIENTS AND METHODS: The principal data were all hospital discharges for New York resident children admitted to hospitals in New York, Pennsylvania, New Jersey, or Connecticut in 1994. The data and methodology were noteworthy for including out-of-area hospital admissions. RESULTS: There was a substantial negative association of the ACS rate with private HMO coverage. There also were sizable negative effects of the availability of primary care services in physician offices and the distance traveled. Large differences related to racial and ethnic composition of the population were found independent of other determinants. There was a positive association with the proportion of all admissions (admissions for all conditions) covered by Medicaid or self-pay. Severity of illness and use of emergency departments were controlled. There was no independent effect of a location in New York City. CONCLUSIONS: The results are consistent with smaller-scale studies suggesting that improved health insurance for children could reduce hospital admissions. Contracting with HMOs also appears to be beneficial. Independently, programs to increase the availability of primary and preventive services could substantially reduce ACS admissions. PMID- 11388128 TI - Impact of HMO ownership on management processes and utilization outcomes. AB - OBJECTIVE: To examine the effects of health maintenance organization (HMO) ownership characteristics on selected utilization outcomes and management processes affecting utilization. STUDY DESIGN: We used 1995 HMO data from the American Association of Health Plans. PATIENTS AND METHODS: Using regression analysis, we examined the relation between HMO utilization (hospital discharges, days, and average length of stay; cardiac catheterization procedures; and average cost of outpatient prescriptions) and the structural characteristics of HMOs: ownership type (insurance company, hospital, physician, independent, and national managed care company), HMO size, for-profit status, model type, geographic region, and payer mix. RESULTS: HMO ownership type is significantly associated with medical management processes, including risk sharing by providers, risk sharing by consumers, and other management strategies. Relative to hospital-owned HMOs, insurance company-owned HMOs have fewer hospital discharges, fewer hospital days, and longer lengths of stay. National managed care organization-owned HMOs have fewer cardiac catheterizations and lower average outpatient prescription costs. Independently owned HMOs have more cardiac catheterizations. For-profit HMOs have lower prescription costs. Relative to hospital-owned HMOs, insurance company-owned HMOs are more likely to use hospital risk sharing and provider capitation and less likely to use out-of-pocket payments for hospital use and a closed formulary. National managed care organization-owned HMOs are less likely to use provider capitation, out-of-pocket payments for hospital use, catastrophic case management, and hospital risk sharing. Physician-hospital-owned HMOs are less likely to use catastrophic case management. For-profit HMOs are more likely to use hospital risk sharing and catastrophic case management. CONCLUSION: HMO ownership type affects utilization outcomes and management strategies. PMID- 11388129 TI - Diabetes mellitus in managed care: complications and resource utilization. AB - OBJECTIVES: To implement a diabetes mellitus surveillance system designed to use administrative data and to demonstrate how it can be used by managed care organizations (MCOs) with different administrative data systems to estimate the prevalence of diabetic complications and describe utilization of services in persons with and without complications and comorbidities. STUDY DESIGN, PATIENTS, AND METHODS: We identified individuals with diabetes mellitus in 3 MCOs in 1993 using 4 sources of computerized data records: inpatient, pharmacy, outpatient, and laboratory. The presence of diabetes mellitus complications and cardiovascular comorbidities were determined using diagnostic and procedural codes. Use of healthcare resources by persons with and without complications and comorbidities was determined from computerized administrative data. RESULTS: The most prevalent complication or comorbidity was cardiovascular disease (45%-53%), followed by eye disease (20%-34%), lower extremity disease (8%-20%), and renal disease (3%-6%). The presence of multiple complications was common and ranged from 14% to 34% in the 3 MCO populations. Compared with persons with none, persons with 2 or more complications or comorbidities used moderately more primary care services (1.3-1.9 times more) and markedly more specialty care services (5.8-6.3 times more), emergency department visits (3.3-5.5 times more), and hospitals stays (3.3-11.9 times more). CONCLUSIONS: Diabetic complications were common and had a large impact on patients' use of healthcare services. Within MCOs, administrative databases are useful tools for estimating and monitoring the prevalence of diabetic complications and the use of healthcare resources associated with these complications. PMID- 11388130 TI - Impact of managed MediCal on California family practice programs. AB - BACKGROUND: An important source of patients for California's family practice program is MediCal. During the past 5 years, MediCal has established a variety of capitated managed care plans. OBJECTIVE: To assess the impact of California's managed MediCal program on the state's 38 family practice training programs. DESIGN: A cross-sectional, retrospective descriptive survey. METHODS: A 3-page, 11-question survey was developed by family practice residency directors and staff from the California Academy of Family Physicians, San Francisco. The 38 family practice programs in existence in California in September 1997 were stratified by type of managed MediCal in their county and by type of sponsoring institution- university, county, community based, staff-model health maintenance organization, or managed care system. RESULTS: Of the 38 family practice programs, 27 responded; 19 of 27 programs participated in managed MediCal. The total number of family health center patients, and the percentage of MediCal patients (48%-60%) at family practice programs was similar when stratified by programs with and without managed MediCal and by type of sponsorship. Most programs reported that they were able to compete effectively, although most also reported increased administrative, nursing, and front office costs. Managed MediCal patients were directly assigned to residents in only 3 of 19 programs. CONCLUSIONS: The introduction of managed MediCal has not adversely affected the number of patients cared for in California's family practice programs. Continued vigilance regarding California family practice programs' involvement in managed MediCal, including collection of accurate data on the number of MediCal patients and the financial and educational implications for California's family practice programs, is warranted. PMID- 11388131 TI - Treatment of hypertension in a managed care setting. AB - BACKGROUND: Based on recommendations of the Fifth and Sixth Reports of the Joint National Committee (JNC) on Prevention, Detection, Evaluation, and Treatment of High Blood Pressure, Health Care Plan (now Univera Healthcare) Buffalo, NY, developed a clinical guideline to improve the management of patients with hypertension. To increase awareness and utilization, the guideline was distributed as hard copy reports and made available through our electronic information system. OBJECTIVE: To determine blood pressure (BP) control rates and adherence to guideline recommendations. STUDY DESIGN: Retrospective chart review. PATIENTS AND METHODS: We randomly sampled hypertensive patients seen during 1998 to evaluate hypertension management. Computerized medical and pharmacy records were reviewed for patient demographics, antihypertensive medications, comorbid conditions, and BP readings. Patient assessment was based on antihypertensive regimen and achievement of target BP according to the recommendations of the guidelines (< 140/90 mm Hg for the general population and < 130/85 mm Hg for special populations). In addition, we assessed control rates using traditional Health Plan Employer Data and Information Set (HEDIS) measures (< 140/90 mm Hg). RESULTS: Overall, 35% of patients achieved target BP and 68% were treated with agents recommended by our JNC-based guideline. In contrast, using traditional HEDIS measures, 41% of patients achieved BP control. Of 39 patients with compelling indications (primarily diabetic patients), 13% achieved BP target and 67% were treated with recommended agents. CONCLUSIONS: The impact of our clinical guideline is reflected through the relatively high utilization of recommended drugs. However, optimal BP control continues to be problematic. In particular, patients with diabetes warrant focused attention. PMID- 11388132 TI - Total cost comparison of 2 biopsy methods for nonpalpable breast lesions. AB - OBJECTIVE: To identify, quantify, and compare total facility costs for 2 breast biopsy methods: vacuum-assisted biopsy (VAB) and needle-wire-localized open surgical biopsy (OSB). STUDY DESIGN: A time-and-motion study was done to identify unit resources used in both procedures. Costs were imputed from published literature to value resources. A comparison of the total (fixed and variable) costs of the 2 procedures was done. PATIENTS AND METHOD: A convenience sample of 2 high-volume breast biopsy (both VAB and OSB) facilities was identified. A third facility (OSB only) and 8 other sites (VAB only) were used to capture variation. Staff interviews, patient medical records, and billing data were used to check observed data. One hundred and sixty-seven uncomplicated procedures (71 OSBs, 96 VABs) were observed. Available demographic and clinical data were analyzed to assess selection bias, and sensitivity analyses were done on the main assumptions. RESULTS: The total facility costs of the VAB procedure were lower than the costs of the OSB procedure. The overall cost advantage for using VAB ranges from $314 to $843 per procedure depending on the facility type. Variable cost comparison indicated little difference between the 2 procedures. The largest fixed cost difference was $763. CONCLUSIONS: Facilities must consider the cost of new technology, especially when the new technology is as effective as the present technology. The seemingly high cost of equipment might negatively influence a decision to adopt VAB, but when total facility costs were analyzed, the new technology was less costly. PMID- 11388133 TI - Cognitive neuropsychology and functional brain imaging: implications for functional and anatomical models of cognition. AB - We discuss the relations between functional imaging and cognitive neuropsychological research. We begin by elaborating on some of the problems of traditional neuropsychological research, which attempted to provide accounts of cognitive performance at a neural as well as at a functional level of description. The difficulties in making neural-level arguments from neuropsychological data include: problems of associated deficits, problems due to interactive effects between brain regions, problems with analyses based on behavioural syndromes, problems due to the influence of compensatory strategies, and problems in separating damaged from disconnected representations. We discuss how cognitive neuropsychology by-passed many of these problems by emphasising functional rather than neural-level theories, though problems with inferences at the neural-level remain. We then consider the contribution that functional imaging can make to cognitive neuropsychology. Using evidence drawn from studies of language, object recognition and visual attention, we argue that functional imaging complements cognitive neuropsychology by: (i) not being reliant on accidents of nature and by enabling effects of lesions on 'distant' neural areas to be measured, (ii) revealing the brain systems necessary and sufficient for a given task, (iii) providing tests of neural-level models of cognition, and by (iv) providing novel evidence on the mechanisms of functional recovery in patients. In addition to this, imaging studies can contribute directly to functional-level theories, by providing converging evidence on the neural locus of cognition--knowing 'where' can allow new inferences about 'how' a given task is performed. PMID- 11388134 TI - Integrating cognitive psychology, neurology and neuroimaging. AB - In the last decade, there has been a dramatic increase in research effectively integrating cognitive psychology, functional neuroimaging, and behavioral neurology. This new work is typically conducting basic research into aspects of the human mind and brain. The present review features as examples of such integrations two series of studies by the author and his colleagues. One series, employing object recognition, mental motor imagery, and mental rotation paradigms, clarifies the nature of a cognitive process, imagined spatial transformations used in shape recognition. Among other implications, it suggests that when recognizing a hand's handedness, imagining one's body movement depends on cerebrally lateralized sensory-motor structures and deciding upon handedness depends on exact match shape confirmation. The other series, using cutaneous, tactile, and auditory pitch discrimination paradigms, elucidates the function of a brain structure, the cerebellum. It suggests that the cerebellum has non-motor sensory support functions upon which optimally fine sensory discriminations depend. In addition, six key issues for this integrative approach are reviewed. These include arguments for the value and greater use of: rigorous quantitative meta-analyses of neuroimaging studies; stereotactic coordinate-based data, as opposed to surface landmark-based data; standardized vocabularies capturing the elementary component operations of cognitive and behavioral tasks; functional hypotheses about brain areas that are consistent with underlying microcircuitry; an awareness that not all brain areas implicated by neuroimaging or neurology are necessarily directly involved in the associated cognitive or behavioral task; and systematic approaches to integrations of this kind. PMID- 11388135 TI - Beyond localisation: a dynamical dual route account of face recognition. AB - After decades of research the notion that faces are special is still at the heart of heated debates. New techniques like brain imaging have advanced some of the arguments but empirical data from brain-damaged patients like paradoxical recognition effects have required more complex explanations aside from localisation of the face area in normal adults. In this paper we focus on configural face processes and discuss configural processes in prosopagnosics in the light of findings obtained in brain-imaging studies. In order to account for data like paradoxical face recognition effects we propose a dual route model of face recognition. The model is based on the distinction between two separate aspects of face recognition, detection and identification, considered as dynamical and interrelated. In this perspective the face detection system appears as the stronger candidate for face-specific processes. The face identification system on the other hand is part of the object recognition system but derives its specificity in part from interaction with the face-specific detection system. The fact that face detection appears intact in some patients provides us with a possible explanation for the interference of configural processes on feature based identification. PMID- 11388136 TI - Blindsight: the role of feedforward and feedback corticocortical connections. AB - When human subjects suffer from a lesion to the primary visual cortex, they lose all visual percepts in the region of space that corresponds to the site of the lesion. However, they are still capable of responding to stimuli in this region when asked to 'guess' or to execute forced-choice motor commands related to these stimuli. This phenomenon, termed blindsight, is still only partly understood. Here, the possible roles of feedforward and feedback corticocortical connections in the visual brain in the understanding of blindsight are reviewed. What emerges is substantial evidence in favor of the theory that unconscious visuo-motor transformations, as in blindsight, may be executed in an entirely feedforward processing cycle, while visual awareness is critically dependent on feedback connections to the primary visual cortex. PMID- 11388137 TI - Abnormal retinotopic representations in human visual cortex revealed by fMRI. AB - The representation of the visual field in early visual areas is retinotopic. The point-to-point relationship on the retina is therefore maintained on the convoluted cortical surface. Functional magnetic resonance imaging (fMRI) has been able to demonstrate the retinotopic representation of the visual field in occipital cortex of normal subjects. Furthermore, visual areas that are retinotopic can be identified on computationally flattened cortical maps on the basis of positions of the vertical and horizontal meridians. Here, we investigate abnormal retinotopic representations in human visual cortex with fMRI. We present three case studies in which patients with visual disorders are investigated. We have tested a subject who only possesses operating rod photoreceptors. We find in this case that the cortex undergoes a remapping whereby regions that would normally represent central field locations now map more peripheral positions in the visual field: In a human albino we also find abnormal visual cortical activity. Monocular stimulation of each hemifield resulted in activations in the hemisphere contralateral to the stimulated eye. This is consistent with abnormal decussation at the optic chiasm in albinism. Finally, we report a case where a lesion to white matter has resulted in a lack of measurable activity in occipital cortex. The activity was absent for a small region of the visual field, which was found to correspond to the subject's field defect. The cases selected have been chosen to demonstrate the power of fMRI in identifying abnormalities in the cortical representations of the visual field in patients with visual dysfunction. Furthermore, the experiments are able to show how the cortex is capable of modifying the visual field representation in response to abnormal input. PMID- 11388138 TI - The effects of aging on visual memory: evidence for functional reorganization of cortical networks. AB - Recent evidence suggests that the mature human brain is capable of substantial functional reorganization following injury. The fact that the brain retains a great deal of plasticity raises the possibility that cortical reorganization may occur during normal aging. We examined this issue by using positron emission tomography (PET) to measure the brain activity associated with short-term memory for simple visual attributes in young and old observers. A two-interval forced choice procedure was used to measure spatial frequency discrimination thresholds for sine wave gratings presented at different inter-stimulus intervals (ISI). Memory load was manipulated by varying the duration of the ISI and by presenting an irrelevant masking stimulus in the middle of the ISI. Old and young observers performed the experiment equally well. However, the neural systems correlated with good performance differed for the two age groups. The results support the hypothesis that the functional networks that underlie visual memory undergo reorganization during aging. PMID- 11388139 TI - The role of transcranial magnetic stimulation (TMS) in studies of vision, attention and cognition. AB - Transcranial magnetic stimulation (TMS) can be conceptualized as a virtual lesion technique, capable of disrupting organized cortical activity, transiently and reversibly. The technique combines good spatial and temporal resolution and, moreover, because it represents an interference technique, can be said to have excellent functional resolution. The following is a review and discussion of the contribution which TMS has made to the study of vision, attention, development and plasticity and speech and language. PMID- 11388140 TI - fMR-adaptation: a tool for studying the functional properties of human cortical neurons. AB - The invariant properties of human cortical neurons cannot be studied directly by fMRI due to its limited spatial resolution. One voxel obtained from a fMRI scan contains several hundred thousands neurons. Therefore, the fMRI signal may average out a heterogeneous group of highly selective neurons. Here, we present a novel experimental paradigm for fMRI, functional magnetic resonance-adaptation (fMR-A), that enables to tag specific neuronal populations within an area and investigate their functional properties. This approach contrasts with conventional mapping methods that measure the averaged activity of a region. The application of fMR-A to study the functional properties of cortical neurons proceeds in two stages: First, the neuronal population is adapted by repeated presentation of a single stimulus. Second, some property of the stimulus is varied and the recovery from adaptation is assessed. If the signal remains adapted, it will indicate that the neurons are invariant to that attribute. However, if the fMRI signal will recover from the adapted state it would imply that the neurons are sensitive to the property that was varied. Here, an application of fMR-A for studying the invariant properties of high-order object areas (lateral occipital complex--LOC) to changes in object size, position, illumination and rotation is presented. The results show that LOC is less sensitive to changes in object size and position compared to changes of illumination and viewpoint. fMR-A can be extended to other neuronal systems in which adaptation is manifested and can be used with event-related paradigms as well. By manipulating experimental parameters and testing recovery from adaptation it should be possible to gain insight into the functional properties of cortical neurons which are beyond the spatial resolution limits imposed by conventional fMRI. PMID- 11388141 TI - Can neuroimaging really tell us what the human brain is doing? The relevance of indirect measures of population activity. AB - Neuroimaging studies using positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) give an indication towards the localization of mental representations and processes in the human brain. It is not clear to what extent such global measures of neuronal activity, pooling across large populations of neurons, can reveal how certain computations are implemented by the neurons in such population ('computational neuroimaging'). Population activity is related tightly to single-cell activity when all neurons in the population have similar response properties. We describe some evidence from single-cell recordings in monkeys that indicates that neurons with similar response properties are not scattered randomly throughout the visual cortex. Notwithstanding this clustering, populations of nearby neurons are still rather heterogeneous, requiring some prudence in deriving single-cell response properties from population activity. The following review of recent neuroimaging studies of the visual system describes to what degree inferences about computations and representations can be drawn from these studies. PMID- 11388142 TI - Defining the cortical visual systems: "what", "where", and "how". AB - The visual system historically has been defined as consisting of at least two broad subsystems subserving object and spatial vision. These visual processing streams have been organized both structurally as two distinct pathways in the brain, and functionally for the types of tasks that they mediate. The classic definition by Ungerleider and Mishkin labeled a ventral "what" stream to process object information and a dorsal "where" stream to process spatial information. More recently, Goodale and Milner redefined the two visual systems with a focus on the different ways in which visual information is transformed for different goals. They relabeled the dorsal stream as a "how" system for transforming visual information using an egocentric frame of reference in preparation for direct action. This paper reviews recent research from psychophysics, neurophysiology, neuropsychology and neuroimaging to define the roles of the ventral and dorsal visual processing streams. We discuss a possible solution that allows for both "where" and "how" systems that are functionally and structurally organized within the posterior parietal lobe. PMID- 11388143 TI - Visual motion and the human brain: what has neuroimaging told us? AB - Recently, neuroimaging techniques have been applied to the study of human motion perception, complementing established techniques such as psychophysics, neurophysiology and neuropsychology. Because vision, particularly motion perception, has been studied relatively extensively, it provides an interesting case study to examine the contributions and limitations of neuroimaging to cognitive neuroscience. We suggest that in the domain of motion perception neuroimaging has: (1) revealed an extensive network of motion areas throughout the human brain, in addition to the well-studied motion complex (MT+); (2) verified and extended findings from other techniques; (3) suggested extensive top down influences on motion perception; and (4) allowed experimenters to examine the neural correlates of awareness. We discuss these contributions, along with limitations and future directions for the neuroimaging of motion. PMID- 11388144 TI - History and future directions of human brain mapping and functional neuroimaging. AB - It has long been known that there is some degree of localisation of function in the human brain, as indicated by the effects of traumatic head injury. Work in the middle of the 20th century, notably the direct cortical stimulation of patients during neurosurgery, suggested that the degree and specificity of such localisation of function were far greater than had earlier been imagined. One problem with the data based on lesions and direct stimulation was that the work depended on the study of what were, by definition, damaged brains. During the second half of the 20th century, a collection of relatively non-invasive tools for assessing and localising human brain function in healthy volunteers has led to an explosion of research in what is often termed "Brain Mapping". The present article reviews some of the history associated with these tools, but emphasises the current state of development with speculation about the future. PMID- 11388145 TI - Functional imaging during speech production. AB - Physiological studies of speech production have demonstrated that even simple articulation involves a range of specialized motor and cognitive processes and the neural mechanisms responsible for speech reflect this complexity. Recently, a number of functional imaging techniques have contributed to our knowledge of the neuroanatomical and neurophysiological correlates of speech production. These new imaging approaches have the advantage of permitting study of large numbers of normal and disordered subjects but they bring with them a host of new methodological concerns. One of the challenges for understanding language production is the recording of articulation itself. The problems associated with measuring the vocal tract and measuring the neural activity during overt speech are reviewed. It is argued that advances in understanding fundamental questions such as what are the planning units of speech, what is the role of feedback during speech and what is the influence of learning, await the development of better methods for assessing task performance. PMID- 11388146 TI - Practical pediatric interventional radiology. PMID- 11388147 TI - [Determination by flow cytometry of reference values of erythrocyte parameters in aged subjects]. AB - OBJECTIVES: Normal values for hemoglobin and hematocrit have been established for healthy young adults, but it is unknown whether age-related decline in hematological values occurs in the elderly. We compared normal values of hematological parameters in healthy adults and healthy persons aged 65 years and over using flow cytometry. SUBJECTS AND METHODS: Erythrocyte and reticulocyte parameters were determined in 585 subjects (294 men and 206 women aged 26 to 64 years, and 37 men and 48 women aged 65 years and over), living independently and without medication, symptom and disease, using Technicon H1 (Bayer) and R2000 (Roche) analyzers. Serum erythropoietin levels, red cell folate and creatinine clearance were determined in subjects aged 65 years and over. RESULTS: Irrespective of subject age, hemoglobin (Hb), hematocrit (Hct), red blood cell count (RBC), mean cell volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) were higher in males than in females, but sex-related differences tended to decrease with aging. In males, there was a trend toward lower Hb and Hct with aging, due to an increase in RBC, MCV and MCH. In females, Hb and Hct (p < 0.05) increased with aging, due to a trend toward higher values of RBC, MCV and MCH. Red blood cell distribution width and hemoglobin distribution width were not modified by aging. Despite these age related changes in erythrocyte parameters, values for anemia defined in adults appear appropriate for geriatric subjects (Hb < 130 g/l in males and Hb < 120 g/l in females). Reticulocyte count and creatinine clearance decreased in men and women aged 65 years and over (p < 0.05), whereas erythropoietin levels and red cell folate were not influenced by aging. CONCLUSION: Normal erythrocyte values defined in adults are appropriate for evaluating hematological test results in older individuals. The decrease in reticulocyte count observed in men and women aged 65 years and over does not appear to be linked to a decrease in erythropoietin production with aging. PMID- 11388148 TI - [Medication error rate in the hospital setting: a pilot study at the Jury-les Metz Hospital Center]. AB - OBJECTIVES: Medication error rate (TEM) is seen as the performance rate of the process leading from drug prescription to drug administration. Jury-les-Metz Psychiatric Hospital conducted a pilot study whose aim was to determine feasibility of drug error rate calculation. METHODS: TEM was calculated as the number of errors divided by the number (n) of all the doses ordered. Four different steps of the process were studied with a specific rate for each step: drug prescription error rate (TEMp), drug delivery calculation error rate (TEMr), drug missing before administration rate (TEMm) and nursing error in drug preparing rate (TEMc). RESULTS: TEMp (n = 15,699): 1.92% [1.74-2.10], TEMr (n = 3457): 2.7% [2.2-3.2], TEMm (n = 3457): 6.9% [6.1-7.7], TEMc (n = 956): 5.9% [4.4 7.4]. DISCUSSION: The research topic presented here is quite new. In spite of certain limitations, the most important point is to focus attention on four specific steps and not to limit the study to the entire process. Our pilot work demonstrates that drug errors are made and must not be neglected. The TEM is not simply the sum of specific errors because of the interactions between the underlying causes of the different types of errors, each affecting the value of specific TEMs. CONCLUSION: Before to deal with quality improvement of hospital medication process, we had to prove that drug error exists. Now, two solve the problem of medication error in our hospital, we have to observe what exactly happens and to try to multidisciplinary understand the causes leading to process defects. PMID- 11388149 TI - [Type 1 membranoproliferative glomerulonephritis in a patient with acquired partial lipodystrophy]. PMID- 11388150 TI - [Carpal tunnel syndrome after occupational use of a motorcycle]. PMID- 11388151 TI - [Cancer of the rectum: contribution of MRI in predicting complete resection]. PMID- 11388152 TI - [Practice Guideline for antibiotic prophylaxis in surgery. SFAR. French Society of Anesthesiology and Resuscitation]. PMID- 11388153 TI - [Reading as a comforting tool]. PMID- 11388154 TI - [Management of therapeutic failure in HIV-infected patients]. AB - A COMMON SITUATION: Among HIV-infected patients treated with antiretroviral regimens, 20% to 50% escape therapeutic control. The principal factors predictive of treatment failure are low CD4 counts and high viral load prior to institution of the antiretroviral treatment. Several virological and pharmacological mechanisms are implicated. GENOTYPING AND PHENOTYPING: Genotyping is particularly useful to optimize treatment in HIV-infected patients who escape therapeutic control, irrespective of the therapeutic strategy. A committee of experts has developed a scheme for adapting treatment to the very complex results of genotyping tests. The benefit of phenotyping remains to be established. MANAGEMENT: Serum concentrations of antiretroviral agents should be determined and adapted in order to maximize the treatment effect. However, to manage therapeutic escape, the only method with a proven impact is genotyping. The possible usefulness of regular drug assays for adjusting treatment doses in case of therapeutic escape remains to be assessed in prospective trials. PMID- 11388155 TI - [Disseminated strongyloidiasis]. AB - ENDEMIC ZONES: Strongyloidiasis is an intestinal parasitosis which is frequently found in tropical and subtropical regions. RISK: The "autoinfection" cycle during this helminthiasis explains why the infection can be perpetuated without further exposure to exogenous, infective larvae. Hyperinfection may occur by dissemination of Strongyloides stercoralis in immuno-compromised patients, particularly those under corticotherapy. DISSEMINATATED STRONGYLOIDIASIS: Disseminated strongyloidiasis is characterized by severe gastrointestinal and respiratory tract involvement, meningitis, skin rash, or Gram-negative bacteremia. Since disseminated stronglyoidiasis is fatal in 80% of cases it is imperative to diagnose and treat this condition before long-term corticotherapy. Ivermectin is currently recommended because it is effective and well tolerated. PMID- 11388156 TI - [Cystic pyelo-ureteritis]. PMID- 11388157 TI - Advancing quality patient care through research. PMID- 11388158 TI - Nurse/physician collaboration: action research and the lessons learned. AB - OBJECTIVE: Finding time to add to nursing knowledge while solving problems in a fast-paced healthcare environment is the ultimate challenge for nurse executives. At one hospital, use of an action research model to measure collaboration in nurse/physician led interdisciplinary teams improved the intervention and the approach to outcome measurement. BACKGROUND: Many hospital nurse executives promote collaborative practice, and yet, innovations introduced to foster collaboration are rarely studied prospectively. The best-known data on collaboration is predominantly from correlational studies. Within the rapidly changing practice setting, action research may be a more legitimate strategy for studying interventions longitudinally. METHODS: An action research pretest/posttest design using Baggs' Collaboration and Satisfaction About Care Decisions measured collaboration before and after several interventions to improve nurse/physician collaboration. The sample consisted of 87 pretest and 65 posttest registered nurses working on three medical-surgical units and two intensive care units (ICU). RESULTS: Collaboration scores in the ICUs were higher than those in previous research, but the posttest indicated no significant difference in either ICU nurse or medical-surgical nurse scores. Higher ICU scores may have been related to the organizational focus on teams. A strong significant correlation between nurse report of level of collaboration and satisfaction with decision making was uncovered. CONCLUSIONS: This study contributes to the nurse/physician collaboration literature in that it was longitudinal, used a reliable and valid instrument, and surveyed nurses in medical/surgical units as well as the ICU. Some of the difficulties and benefits of research in today's practice setting are illustrated. PMID- 11388159 TI - Testing Karasek's Demands-Control Model in restructured healthcare settings: effects of job strain on staff nurses' quality of work life. AB - BACKGROUND: Job strain among staff nurses has become an increasingly important concern in relationship to employee performance and commitment to the organization in current restructured healthcare settings. OBJECTIVES: The purpose of this study was to test Karasek's Demands-Control Model of job strain by examining the extent to which the degree of job strain in nursing work environments affects staff nurses' perceptions of structural and psychological empowerment, work satisfaction, and organizational commitment. METHOD: A predictive, nonexperimental design was used to test these relationships in a random sample of 404 Canadian staff nurses. Karasek's Job Content Questionnaire, the Conditions of Work Effectiveness Questionnaire-II, Spreitzer's Psychological Empowerment Questionnaire, Meyer and Allen's Organizational Commitment Questionnaire, and the Global Satisfaction Scale were used to measure the major study variables. RESULTS: Nurses with higher level of job strain were found to be significantly more empowered, more committed to the organization, and more satisfied with their work. CONCLUSIONS: Support for Karasek's Demands/Control theory was established in this study. PMID- 11388160 TI - Effect of locale on health status and direct care time of rural versus urban home health patients. AB - OBJECTIVE: The study purpose was to determine if health status differed between rural versus urban home health patients and to identify if locale was a significant predictor of home health direct care time. BACKGROUND: Implemented in October 2000, the Medicare home health prospective payment system (PPS) made only temporary allowance for differences in cost of delivering home health services in rural versus urban environments. However, past research documented differences in health status and service utilization between rural and urban home health patients. METHODS: Data were collected retrospectively on a convenience sample of 2,788 patient episodes of care. Patient health status was measured using items form the Outcome Assessment and Information Set (OASIS). Obtained from itinerary records, direct care time was the time clinicians spent in the home. RESULTS: The results showed significant differences in rural versus urban patients health status, with urban patients being healthier than rural patients. Consistent with poorer health status, rural patients received more RN direct care time. Other study factors being equal, living in a rural locale increased total direct care time by 150 minutes over living in an urban environment. CONCLUSIONS: Given the poorer health status and increased time requirements for rural patients, the results support assertions that Medicare per episode reimbursement should be higher for rural than for urban home health patients. PMID- 11388161 TI - Q methodology: a new way of assessing employee satisfaction. AB - As yet another nursing shortage faces the country, the issue of the satisfaction of nurses again becomes of critical concern to nursing managers in the interest of staff retention. The authors describe the use of the statistical technique Q methodology to assess the needs of nurses and other medical staff at a level one, tertiary care emergency department in the United States. Using the Q method, the authors were able to identify different, unique viewpoints concerning employee needs among the study population, as well as commonly shared views. This level of detail, not obtainable using more traditional statistical techniques, can aid in the design of more effective strategies aimed at fulfilling the needs of an organization's staff to increase their satisfaction. PMID- 11388162 TI - Impact of structural and psychological empowerment on job strain in nursing work settings: expanding Kanter's model. AB - OBJECTIVE: In this study, we tested an expanded model of Kanter's structural empowerment, which specified the relationships among structural and psychological empowerment, job strain, and work satisfaction. BACKGROUND: Strategies proposed in Kanter's empowerment theory have the potential to reduce job strain and improve employee work satisfaction and performance in current restructured healthcare settings. The addition to the model of psychological empowerment as an outcome of structural empowerment provides an understanding of the intervening mechanisms between structural work conditions and important organizational outcomes. METHODS: A predictive, nonexperimental design was used to test the model in a random sample of 404 Canadian staff nurses. The Conditions of Work Effectiveness Questionnaire, the Psychological Empowerment Questionnaire, the Job Content Questionnaire, and the Global Satisfaction Scale were used to measure the major study variables. RESULTS: Structural equation modelling analyses revealed a good fit of the hypothesized model to the data based on various fit indices (chi 2 = 1140, df = 545, chi 2/df ratio = 2.09, CFI = 0.986, RMSEA = 0.050). The amount of variance accounted for in the model was 58%. Staff nurses felt that structural empowerment in their workplace resulted in higher levels of psychological empowerment. These heightened feelings of psychological empowerment in turn strongly influenced job strain and work satisfaction. However, job strain did not have a direct effect on work satisfaction. CONCLUSIONS: These results provide initial support for an expanded model of organizational empowerment and offer a broader understanding of the empowerment process. PMID- 11388163 TI - How to host a nursing symposium, from planning to cleanup. PMID- 11388164 TI - [Platon Ivanovich Tikhov (1865-1917)]. PMID- 11388165 TI - Leiomyoma of the urethra in a Mexican woman: a rare neoplasm associated with the expression of estrogen receptors by immunohistochemistry. AB - BACKGROUND: Leiomyoma of the female urethra is a rare condition, although it can occur anywhere along the genitourinary tract. METHODS: We report on a 22-year-old woman found to have a urethral mass detected in our hospital delivery room. Examination showed a 6 X 5-cm mass at the urethral meatus. RESULTS: Pathologic examination revealed urethral leiomyoma. Immunohistochemistry confirmed leiomyoma with positive staining for vimentin, desmin, and actin. Immunoreactivity for estrogen receptors was also detected. CONCLUSIONS: Because this lesion was discovered while the patient was pregnant and it showed immunoreactivity for estrogen receptors, it is suggested that increased estrogen levels could accelerate smooth muscle growth. PMID- 11388166 TI - Web alert. PMID- 11388167 TI - [Radon exposure and risks for public health]. PMID- 11388168 TI - The panmalarial antigen detected by the ICT Malaria P.f/P.v immunochromatographic test is expressed by Plasmodium malariae. PMID- 11388169 TI - Changes in Campylobacter jejuni and Campylobacter coli carriage rates in the Zenica Region [correction of Zenica Canton] of Bosnia and Herzegovina in the pre- and postwar periods. PMID- 11388170 TI - Discordant carbapenem susceptibility in Methylobacterium species and its application as a method for phenotypic identification. PMID- 11388171 TI - Recombinant low-molecular-mass proteins pG and LA7 from Borrelia burgdorferi reveal low diagnostic sensitivity in an enzyme-linked immunosorbent assay. PMID- 11388173 TI - Bibliography. Current world literature. Epidemiology and prevention. PMID- 11388172 TI - Bibliography. Current world literature. Renal immunology and pathology. PMID- 11388174 TI - Bibliography. Current world literature. Renal pathophysiology. PMID- 11388177 TI - Guiding principles for research involving animals and human beings. PMID- 11388175 TI - Reply to letter to editor by M.J. Sankeralli and K.T. Mullen published in Vision Research, 41, 53-55: Lights and neural responses do not depend on choice of color space. PMID- 11388178 TI - Lottery of NHS funding is inappropriate. PMID- 11388179 TI - Doctors should not advise adolescents to abstain from sex. PMID- 11388180 TI - Emergency contraception from pharmacists misses opportunity. PMID- 11388181 TI - Psychosocial impacts of chlamydia testing are important. PMID- 11388182 TI - Glycaemia and vascular effects of type 2 diabetes. Lowering glucose concentrations may not be of any value in itself. PMID- 11388183 TI - Glycaemia and vascular effects of type 2 diabetes. Relation between diabetes and hyperglycaemia and cardiovascular disease has not been resolved. PMID- 11388184 TI - Glycaemia and vascular effects of type 2 diabetes. UKPDS is not a cohort study and analysis is misleading. PMID- 11388185 TI - Genetic contribution to osteoarthritis of the hip. Did ethics committee consult specialists. PMID- 11388186 TI - Genetic contribution to osteoarthritis of the hip. Genetic contribution needs further investigation. PMID- 11388187 TI - In praise of mercury sphygmomanometers. Appropriate sphygmomanometer should be selected. PMID- 11388188 TI - In praise of mercury sphygmomanometers. Electronic readings of blood pressure seem to be higher than readings obtained with mercury sphygmomanometers. PMID- 11388189 TI - Marketing studies and scientific research must be distinct. PMID- 11388190 TI - All NHS consultants must have equal entitlement to awards. PMID- 11388191 TI - Carpentier-Edwards supraannular porcine bioprosthesis: second-generation prosthesis in aortic valve replacement. AB - BACKGROUND: The Carpentier-Edwards supraannular porcine bioprosthesis experience for more than 18 years has been evaluated by actuarial and actual analysis to determine the clinical performance in aortic valve replacement. METHODS: From 1981 to 1998, 1,823 patients (mean age 68.5 years, range 20 to 90 years) underwent 1,846 procedures. Previous coronary artery bypass was performed in 3.1% (56) and previous valve repair/replacement in 6.0% (110). Concomitant coronary artery bypass grafting was performed in 41.5% (756). RESULTS: The overall valve related complication rate was 4.5%/patient-year (567 patients) with a fatality rate of 0.9%/patient-year (110 patients). The patient survival, at 15 years, was 33.0%+/-3.7% for the 61 to 70 years age group and 13.5%+/-2.4% for the older than 70 years group. At 15 years, the overall actual, cumulative freedom from reoperation was 83.2%+/-1.4%, valve-related mortality was 88.0%+/-1.2%, and valve related residual morbidity was 92.0%+/-0.8%. The actual freedom from structural valve deterioration at 15 years was 84.2%+/-2.8% for the 61 to 70 years group and 97.1%+/-0.9% for the older than 70 years group. CONCLUSIONS: The Carpentier Edwards porcine bioprosthesis provides excellent freedom from structural valve deterioration, and overall freedom from valve-related morbidity, mortality, and reoperation for aortic valve replacement for up to 15 years. The prosthesis is recommended for patients older than 70 years and for patients 61 to 70 years, especially when extended survival is not anticipated. PMID- 11388192 TI - Carpentier-Edwards supraannular porcine bioprosthesis in aortic position: 16-year experience. AB - BACKGROUND: The aim of the study was to evaluate the long-term results of aortic valve replacement with the Carpentier-Edwards supraannular porcine bioprosthesis. METHODS: A total of 278 patients who underwent aortic valve replacement between January 1983 and December 1986 were reviewed. Mean age was 69.4+/-11.0 years (range 24 to 90 years). RESULTS: The operative mortality was 8.6% (24 patients). The total follow-up was 2367.1 years (mean 9.3+/-4.3 years, maximum 15.5). The late mortality rate was 6.8%/patient-year (162 patients) and the overall survival at 15 years was 26.5%+/-3.6%. Structural valve deterioration (SVD) occurred in 19 patients (linearized rate 0.8%/ patient-year). The mean time to onset of deterioration was 10.9+/-2.9 years. This time was independent of the age at the time of implantation. The freedom from SVD at 10, 12, and 15 years for patients aged less than 60 was respectively 87.6%+/-6.8%, 77.8%+/-8.9%, and 44.2%+/-12.9% (linearized rate 3.3%/patient-year). For patients aged 61 to 70 years, freedom from SVD was, respectively, 100%, 97.3%+/-2.1%, and 80.8%+/-8.3% (linearized rate 0.63% patient-year). For patients older than 70 years, it was respectively 99.1%+/-0.9%, 95.6%+/-2.6%, and 93.3%+/-3.3% (linearized rate 0.31%/patient year). No significant difference was observed below the age of 60 years (< or =50 vs 51 to 60 years) or in the older subgroups (61 to 70 years, vs >70 years). CONCLUSIONS: The Carpentier-Edwards supraannular bioprosthesis in aortic position provides low rate of structural valve deterioration at 15 years in patients aged more than 60 years at the time of implantation. The mean time to onset of SVD is independent of the subject's age at the time of implantation. After 60 years, the risk of deterioration is low and does not present any significant variation. The Carpentier-Edwards supraannular bioprosthesis can reliably be used for aortic valve replacement in patients over the age of 60 years because, beyond this age, SVD is observed much more rarely. PMID- 11388193 TI - Over twenty-year follow-up of the standard Hancock porcine bioprosthesis implanted in the mitral position. AB - BACKGROUND: To define the long-term results of 331 standard Hancock porcine bioprostheses implanted in the mitral position between 1973 and 1980. METHODS: Of 331 patients (225 male patients, 68%), mean age 49+/-10 years (range 14 to 69 years), 88% were in New York Heart Association functional class III or IV and 77% were in atrial fibrillation. Follow-up time extended more than 20 years (mean 13.9 years, range 0.3 to 24.7 years) for a total of 4,601 patient-years. RESULTS: Overall operative mortality was 6.3%. At 5, 10, 15, and 20 years, the actuarial survival rate of patients were 71%+/-2%, 46%+/-3%, 30%+/-3%, and 22%+/-2%, respectively. Actuarial estimates of freedom from structural valve deterioration were 95%+/-1%, 67%+/-3%, 32%+/-3%, and 14%+/-3%; from reoperation were 96%+/-1%, 72%+/-3%, 36%+/-4%, and 18%+/-4%; from thromboembolism were 89%+/-2%, 82%+/-3%, 74%+/-4%, and 51%+/-2%; and from anticoagulant-related hemorrhage were 98%+/-1%, 96%+/-1%, 91%+/-1%, and 86%+/-4%. Estimates of freedom from all valve-related mortality at 5, 10, 15, and 20 years were 89%+/-2%, 76%+/-3%, 64%+/-4%, and 48%+/ 4%. Multivariate analysis showed younger age to be a significant risk factor for reoperation. Age at operation did not correlate with structural valve deterioration. CONCLUSIONS: The long-term results with the standard Hancock bioprosthesis implanted in the mitral position appear satisfactory, particularly up to 15 years from implantation. Protection from stroke, anticoagulant hemorrhage, and endocarditis was good. PMID- 11388194 TI - Fifteen-year experience with the mitral Carpentier-Edwards PERIMOUNT pericardial bioprosthesis. AB - BACKGROUND: This multicenter study concerning the mitral PERIMOUNT valve previously reported clinical results at 12 years; this report updates the performance to 15 years postoperatively. METHODS: The 435 patients (mean age 60.7+/-11.6 years; 41.1% male) underwent implantation with the PERIMOUNT valve between 1984 and 1989 at seven institutions. Follow-up was complete for 96.1% of the cohort. The mean follow-up was 8.1+/-4.4 years (range 0 to 15.4 years) for a total of 3492 patient-years. RESULTS: There were 34 (7.8%) operative deaths, one (0.2%) valve related. The late mortality rate was 5.3%/patient-year (2.2%/patient year valve related). At 14 years, the overall actuarial survival rate was 37.1%+/ 3.3% (63.1%+/-4.4% valve related). Actuarial freedom from complications at 14 years was as follows: thromboembolism, 83.8%+/-3.2% (1.1%/patient-year); hemorrhage, 86.6%+/-3.2% (1.1%/patient-year); and explant due to structural valve deterioration (SVD), 68.8%+/-4.7%. Actual freedom from explant due to SVD was 83.4%+/-2.3%. Rates of structural failure decreased with increasing age at implant. CONCLUSIONS: The Carpentier-Edwards PERIMOUNT Pericardial Bioprosthesis is a reliable choice for a tissue valve in the mitral position, especially in patients more than 60 years of age. PMID- 11388195 TI - Comparison of durability of bioprostheses in tricuspid and mitral positions. AB - BACKGROUND: Few reports have assessed differences in the durability of mitral and tricuspid bioprostheses after simultaneous implantation of the same bioprosthesis in both positions. We investigated the long-term outcome after simultaneous mitral valve replacement (MVR) and tricuspid valve supraannular implantation (TVSI) with the Carpentier-Edwards bioprostheses in patients with severe tricuspid regurgitation and advanced mitral valve disease. METHODS: Between 1982 and 1998, 37 patients in our hospital underwent MVR and TVSI with Carpentier Edwards bioprostheses. The mean age of the patients was 55+/-11 years. The average postoperative follow-up was 7.9+/-4.5 years after surgery (range 0 to 14.6 years, 315.1 patient-years). The follow-up rate was 100%. We evaluated the actuarial survival rate, the actuarial freedom from structural valve deterioration (SVD) and reoperation, and postoperative complications. RESULTS: The overall actuarial survival rate at 13 years after the operation was 69%+/ 31%. The actuarial freedom from SVD and reoperation in the mitral and tricuspid positions were 78+/-22 and 100% and 70+/-30 and 90%+/-10% (p = 0.03), respectively. No patient had systemic or pulmonary thromboembolism, or complications associated with fatal arrhythmia. CONCLUSIONS: These results suggest that the bioprostheses in the tricuspid position yield significantly better long-term results than those in the mitral position after simultaneous MVR and TVSI. PMID- 11388196 TI - Long-term results of the bioprosthesis in elderly patients: impact on quality of life. AB - BACKGROUND: A wealth of data exists on acceptable mortality and morbidity for valve operations in older patients, yet information documenting quality of life is lacking. METHODS: From October 1974 to May 1998, 2,075 patients aged 65 years and older underwent valve replacement using a porcine bioprosthesis. There were 1,126 men (54.3%) and 949 women (45.7%) with a mean age of 73.9 years (range 65 to 104 years). RESULTS: The elective hospital mortality was 8.5% (158 patients), and urgent/emergent/salvage mortality was 25.8% (54 patients). Follow-up was completed for 1,863 patients (98.2%) and extended from 1 month to 23.0 years (mean 60.8 months) with a cumulative follow-up of 9,442.1 patient-years. At follow-up, surviving patients (n = 849) completed the Short Form-36 Quality of Life Survey. Results showed patients had a more favorable quality of life compared with control subjects matched for age and sex. Functional improvement was significant with 96.3% in New York Heart Association functional class I or II at follow-up. There were 74 valves that failed from all causes (33 aortic and 41 mitral valves). Actuarial freedom from valve failure at 9 years was 94.4%+/-1.1% and at 18 years was 83.7%+/-2.4%. CONCLUSIONS: Valve replacement in older patients provides excellent functional improvement, reduces late cardiac events, and enhances quality of life. PMID- 11388198 TI - Aortic valve replacement with mechanical and biologic prosthesis in middle-aged patients. AB - BACKGROUND: Mechanical prostheses are used in young patients and bioprostheses in the elderly because of the higher rate of structural failure of bioprostheses. The objective of the present study was to compare results after aortic valve replacement with mechanical (Carbo-Medics) and biologic (Carpentier-Edwards pericardial) in middle-aged patients. METHODS: Five hundred twenty-one patients, aged between 55 and 65 years, who underwent aortic valve replacement with mechanical (n = 363) or biologic (n = 158) prostheses were reviewed. RESULTS: The 10-year actuarial survival rate averaged 66%+/-6% in patients implanted with mechanical valves compared with 75%+/-4% in patients implanted with biologic valves (p = 0.2). The 10-year freedom rate from thromboembolism, hemorrhage, and endocarditis averaged 92%+/-7%, 97%+/-2%, and 99%+/-1%, respectively, in patients with mechanical valves compared with 91%+/-3% (p = 0.03), 99%+/-1% (p = 0.4), and 95%+/-2% (p = 0.01), respectively, in those with biologic valves. The 10-year freedom rate from all valve-related complications averaged 90%+/-7% and 83%+/-4%, respectively (p = 0.01). CONCLUSIONS: The freedom rate from all valve-related complications was higher among patients with mechanical valves compared with biologic valves 10 years after aortic valve replacement in middle-aged patients. PMID- 11388197 TI - Experience with the 19-mm Carpentier-Edwards pericardial bioprosthesis in the elderly. AB - BACKGROUND: Valve replacement in small aortic root remains a surgical challenge. The objective of this study was to compare results of the 19-mm bioprosthesis with those of larger prostheses in the elderly. METHODS: The 443 patients, 70 years of age and older, who underwent aortic valve replacement with Carpentier Edwards pericardial bioprostheses were reviewed. RESULTS: There were 93 patients with a mean age of 76+/-4 years with implantation of 19-mm prostheses and 350 patients with a mean age of 75+/-4 years with larger bioprostheses. Associated aortoplasty was performed in 10 patients (11%) with 19-mm bioprostheses and in 8 patients (2%) with larger bioprostheses (p = 0.001). There were 11 deaths (12%) within 30 days of surgery in patients with 19-mm prostheses and 22 deaths (6%) among those with larger prostheses (p = 0.1). The 7-year survival rate averaged 61%+/-7% in patients with 19-mm prostheses and 67%+/-4% in those with larger prostheses (p = 0.8). The 7-year freedom rates from all valve-related events averaged 96%+/-2% and 93%+/-2%, respectively (p = 0.6). CONCLUSIONS: Aortic valve replacement with the 19-mm Carpentier-Edwards pericardial bioprosthesis offers excellent midterm results in the elderly. PMID- 11388199 TI - Mechanical or bioprosthetic valves in the elderly: a 20-year comparison. AB - BACKGROUND: Our objective was to compare long-term results of mechanical and bioprosthetic valve replacement in patients older than 70 years. METHODS: Patients older than 70 years who had either a St. Jude Medical (SJM) mechanical prosthesis or any bioprosthesis (BP) implanted between January 1977 and December 1997 were identified. Alive patients were interviewed by telephone during a closing interval of 130 days. RESULTS: Complete follow-up was achieved with a total follow-up of 2,264 patient years. A total of 547 patients had 448 aortic valve replacements (199 SJM and 249 BP) and 99 had mitral valve replacements (76 SJM and 23 BP). A further 30 patients had double valve replacement. One hundred ninety of the 577 patients (33%) had coronary artery bypass grafting in addition to the valve replacement. Survival analysis showed no advantage for either mechanical or bioprosthetic valves. There was also no difference in thromboembolic rates, paravalvular leaks, structural dysfunction, and endocarditis rates. However, patients with mechanical valves had a significantly greater risk of major (p < 0.0001) and minor bleeding (p = 0.002) events. CONCLUSIONS: Bioprosthetic valves do not offer a survival advantage over mechanical valves among the elderly. However, anticoagulant-related mortality and morbidity is statistically higher for patients with mechanical valves. PMID- 11388200 TI - Late results of double-valve replacement with biologic or mechanical prostheses. AB - BACKGROUND: We previously showed that the risk of reoperation for structural degeneration of bioprostheses was higher in cases involving patients older than 65 years (p = 0.003) and double-valve replacement (p = 0.02). The purpose of this study was to compare late outcome of mitral-aortic valve replacement using bioprostheses or mechanical valves. METHODS: The bioprosthesis group included all mainland France residents (n = 48) between 55 and 65 years old operated on between 1980 and 1995 for mitral-aortic valve replacement using bioprostheses. The mechanical valve group was obtained by matching each of these patients with a patient operated on using mechanical valves at approximately the same time during the study. RESULTS: In the bioprosthesis group, 10-year survival was 45%+/-8% versus 62%+/-7% in the mechanical valve group (not significant). The linearized reoperation rate was 6.8 per patient-year versus 1.1 per patient-year (p = 0.001), and the linearized reoperative mortality rate was 1.8 per patient-year and 0.7 per patient-year (not significant), respectively. CONCLUSIONS: The reoperative mortality risk after mitral-aortic valve replacement using two bioprostheses does not significantly decrease overall survival after age 65 years. PMID- 11388201 TI - Patient-prosthesis mismatch can be predicted at the time of operation. AB - BACKGROUND: Patient-prosthesis mismatch is a frequent cause of high postoperative gradients in normally functioning prostheses. The objective of this study was to determine whether mismatch can be predicted at the time of operation. METHODS: Indices used to predict mismatch were valve size, indexed internal geometric area, and projected indexed effective orifice area (EOA) calculated at the time of operation, and results were compared with indexed EOA and mean gradients measured by Doppler echocardiography after operation in 396 patients. RESULTS: The sensitivity and specificity of these indices to detect mismatch, defined as a postoperative indexed EOA of 0.85 cm2/m2 or less, were respectively: 35% and 84% for valve size, 46% and 85% for indexed internal geometric area, and 73% and 80% for projected indexed EOA. Projected indexed EOA also correlated best with resting (r = 0.67) and exercise (r = 0.77) postoperative gradients. CONCLUSIONS: The projected indexed EOA calculated at the time of operation accurately predicts mismatch as well as resting and exercise postoperative gradients, whereas valve size and indexed internal geometric area cannot be used for this purpose. PMID- 11388202 TI - Medtronic Mosaic porcine bioprosthesis: midterm investigational trial results. AB - BACKGROUND: The midterm clinical and hemodynamic performance of the Medtronic Mosaic porcine bioprosthesis was evaluated in a regulatory trial. METHODS: In nine Canadian centers, 802 bioprostheses (560 aortic and 242 mitral) were implanted between September 1994 and April 1999 in patients with a mean age of 70 years. RESULTS: Survival for aortic valve replacement at 4 years was 84.4%+/ 3.1%. Freedom from valve-related or unexplained death was 95.6%+/-1.9%; structural valve deterioration, 100.0%; reoperation, 96.2%+/-1.7%; major thromboembolism, 96.1%+/-1.8%; and major antithrombotic-related hemorrhage, 96.4%+/-1.7%. Echocardiographic derived mean systolic gradient was 13.4 mm Hg at 4 years with an indexed effective orifice area of 0.7 to 0.8 cm2/m2. A significant decrease in left ventricular mass was shown over time in all valve sizes. Survival for mitral valve replacement at 4 years was 79.2%+/-6.8%. Freedom from valve-related or unexplained death was 96.5%+/-3.4%; structural valve deterioration, 100%; reoperation, 97.0%+/-3.2%; major thromboembolism, 95.7%+/ 3.8%; and major antithrombotic-related hemorrhage, 95.0%+/-4.2%. Echocardiographically measured averaged mean diastolic gradient was 4.5 mm Hg. CONCLUSIONS: The Medtronic Mosaic bioprosthesis is safe and effective in both the aortic and mitral positions. The valve has low gradients in both positions and excellent left ventricular mass regression in the patients with aortic valve replacement. PMID- 11388203 TI - Mosaic valve international clinical trial: early performance results. AB - BACKGROUND: A new third generation porcine bioprosthesis was developed in an attempt to improve on hemodynamic performance and durability of current prostheses. METHODS: One thousand, two hundred, sixty patients underwent aortic valve replacement and 366 patients underwent mitral valve replacement between February 1994 and September 2000. The cumulative follow-up was 3,696.3 patient years for aortic valve replacement and 880.1 patient-years for mitral valve replacement. Follow-up was complete for 95.5% of aortic valve replacement patients and 97.5% of mitral valve replacement patients. RESULTS: For aortic valve replacement, freedom from valve-related adverse events at 1 year was 96.5%+/-0.5% for antithromboembolic-related hemorrhage and 100% for structural valve deterioration. Freedom from valve-related adverse events at 5 years was 93.8%+/-2.6% for antithromboembolic-related hemorrhage and 99.3%+/-0.9% for structural valve deterioration. For mitral valve replacement, freedom from valve related adverse events at 1 year was 96.0%+/-1.1% for antithromboembolic-related hemorrhage and 100% for structural valve deterioration. Freedom from valve related adverse events at 4 years was 92.1%+/-3.7% for antithromboembolic-related hemorrhage and 100% for structural valve deterioration. CONCLUSIONS: These results support the claim that the Mosaic bioprosthetic valve is efficacious and safe, but continued follow-up is mandatory to determine mid- and long-term performance. PMID- 11388204 TI - Medtronic Intact porcine bioprosthesis experience to twelve years. AB - BACKGROUND: The Medtronic Intact porcine bioprosthesis was further evaluated to determine the influence of zero-pressure glutaraldehyde fixation on structural valve deterioration (SVD). METHODS: From 1986 through 1996, at three Canadian centers, 1,272 patients had 1,296 procedures: 836 aortic valve replacement (AVR), 332 mitral valve replacement (MVR), 14 tricuspid valve replacement, 3 pulmonary valve replacement, and 111 multiple valve replacements. The mean age of the patient population was 67 years (range 9 to 91 years). The total follow-up was 8,011 patient-years (mean 6.2 years). RESULTS: The late mortality (overall) was 4.8% and 6.7% per patient-year for AVR with or without concomitant procedures, respectively; and 4.7% and 10.4% per patient-year for MVR, respectively. There were 51 cases of SVD (AVR 22 of 836; MVR 23 of 332; pulmonary valve replacement 1 of 3; and multiple valve replacement 5 of 111). The actuarial freedom from SVD at 12 years for AVR was 94.3%+/-3.3% for patients aged 61 to 70 years and 97.7%+/ 1.1% for those more than 70 years; for MVR actuarial freedom from SVD at 12 years was 93.7%+/-3.9% for patients more than 70 years. The actual freedom at 12 years from SVD for AVR was 92.4%+/-3.1% for patients aged 51 to 60 years, 96.1%+/-2.1% for those 61 to 70 years, and 98.4%+/-0.7% for those older than 70 years; for MVR actual freedom from SVD at 12 years was 89.6%+/-3.2% for patients 61 to 70 years and 96.6%+/-3.4% for those more than 70 years. CONCLUSIONS: The Medtronic Intact porcine bioprosthesis, formulated with tissue preservation at zero-pressure fixation, has encouraging freedom from structural failure. PMID- 11388205 TI - Hemodynamic comparison of second- and third-generation stented bioprostheses in aortic valve replacement. AB - BACKGROUND: The hemodynamic performance of aortic replacement prostheses is of extreme importance. There is renewed interest in hemodynamics because of the influence of prosthesis-patient mismatch on left ventricular mass regression and the potential influence on survival. METHODS: The hemodynamic performance of the second-generation Carpentier-Edwards supraannular porcine and pericardial (Perimount) bioprostheses and the third-generation Medtronic Mosaic porcine bioprosthesis were compared for mean gradient and effective orifice area index. The effective orifice area index of at least 0.85 cm2/M2 was considered as lack of prosthesis-patient mismatch. The study group included included 53 patients with Carpentier-Edwards supraannular porcine, 48 with pericardial, and 98 with Medtronic Mosaic porcine bioprostheses. RESULTS: The mean gradients were not different between the prostheses by prosthesis size. The Medtronic Mosaic was not provided in size 19. The mean gradients for the prostheses, except in the very large sizes, were all double-digit values. The effective orifice area index was not different between the prostheses but there was a trend toward prosthesis patient mismatch in smaller size prostheses. CONCLUSIONS: There was no apparent hemodynamic advantage between porcine and pericardial bioprostheses in the aortic position. PMID- 11388206 TI - Short-term hemodynamic performance of the mitral Carpentier-Edwards PERIMOUNT pericardial valve. Carpentier-Edwards PERIMOUNT Investigators. AB - BACKGROUND: Although long-term durability data exist, little data are available concerning the hemodynamic performance of the Carpentier-Edwards PERIMOUNT pericardial valve in the mitral position. METHODS: Sixty-nine patients who were implanted with mitral PERIMOUNT valves at seven international centers between January 1996 and February 1997 consented to participate in a short-term echocardiography follow-up. Echocardiographs were collected at a mean of 600+/ 133 days after implantation (range, 110 to 889 days); all underwent blinded core lab analysis. RESULTS: At follow-up, peak gradients were 9.09+/-3.43 mm Hg (mean, 4.36+/-1.79 mm Hg) and varied inversely with valve size (p < 0.05). The effective orifice areas were 2.5+/-0.6 cm2 and tended to increase with valve size (p = 0.08). Trace mitral regurgitation (MR) was common (n = 48), 9 patients had mild MR, 1 had moderate MR, none had severe MR. All MR was central (n = 55) or indeterminate (n = 3). No paravalvular leaks were observed. Mitral regurgitation flow areas were 3.4+/-2.8 cm2 and were without significant volumes. CONCLUSIONS: In this multicenter study, these mitral valves are associated with trace, although physiologically insignificant, central MR. Despite known echocardiographic limitations, the PERIMOUNT mitral valves exhibit similar hemodynamics to other prosthetic valves. PMID- 11388207 TI - Pericarbon pericardial bioprosthesis: an experience based on the lessons of the past. AB - BACKGROUND: The Pericarbon pericardial bioprosthesis, at the time of its creation, showed a breakthrough in terms of low calcification deposit rate, absence of valvular tears, and durability. The purpose of this study was to evaluate results after 10 years. METHODS: From September 1988 to December 1997, 277 patients received a total of 287 Pericarbon pericardial valves. There were 224 (80.8%) isolated aortic valve replacements (AVR), 39 isolated mitral valve replacements (MVR), 1 tricuspid valve replacement, 3 pulmonary valve replacements, and 10 aortic and mitral valve replacements. The total cumulative follow-up was 1,221.42 patient-years (mean 4.9+/-2.6 years). RESULTS: Overall hospital mortality was 10.1%. The overall patient survival at 10.8 years was 55.8%+/-4.2%, for AVR it was 60.0%+/-4.5%, and for MVR it was 46.5%+/-11.9%. The freedom from valve-related death for the overall population at 10.8 years was 98.0%+/-1.0%, for AVR 97.6%+/-1.1%, and for MVR 100%. The overall freedom from structural valve deterioration was 96.6%+/-2.4%, for AVR 96.1%+/-2.7%, and for MVR 100%. The overall freedom from embolic events was 96.0%+/-1.5%, for AVR 96.0%+/-1.6%, and for MVR 100%. The overall freedom from reoperation was 88.1%+/ 3.8%, for AVR 89.9%+/-4.2%, and for MVR 80.6%+/-7.3%. CONCLUSIONS: These results show that over a period of up to 10 years, the Pericarbon pericardial bioprosthesis is an excellent and safe valve substitute. Developing a detoxification process aimed at improving the biological behavior of the glutaraldehyde-tanned valve may increase those advantages. PMID- 11388208 TI - Comparison of three different types of stentless valves: full root or subcoronary. AB - BACKGROUND: It is believed that, compared with stented valves, stentless bioprostheses at the aortic position offer a larger orifice area. METHODS: During the past 45 months, we have implanted 211 various types of aortic prostheses in our clinic. In the subcoronary position, we have used the Medtronic Freestyle, Toronto SPV, and Cryolife O'Brien prostheses, and as an aortic root replacement, the Medtronic Freestyle. There were no special indications for selection of each prosthesis except in 8 patients suffering from a disease of the ascending aorta in addition or in a redo procedure because of endocarditis or valve degeneration in which we implanted the full root Freestyle prosthesis. All patients had echocardiographic examinations postoperatively and after 1 year. RESULTS: Although the implantations took significantly longer time initially, recently the complication rate has shown itself to be no greater than in comparable patients with stented prostheses. The hemodynamic results are very good with the exception of the Freestyle prosthesis implanted in the subcoronary position. The gradients of the remaining three prosthesis after 1 year are between 5 mm Hg and 10 mm Hg, and the effective valve orifice is between 2 and 3 cm2 depending on valve size. CONCLUSIONS: The use of stentless tissue valves offers better hemodynamic results than that of stented valves with essentially no increased operative risk. PMID- 11388210 TI - Clinical outcomes after aortic valve replacement with the Toronto stentless porcine valve. AB - BACKGROUND: Hemodynamic benefits of the Toronto stentless porcine valve have been documented. Clinical well-being and freedom from major valve-related events have been less well defined. METHODS: A total of 447 patients were prospectively followed for up to 8 years (1,745.2 valve years total, 3.9 valve years/patient). The patient demographics included 66% men, mean age 65 years, New York Heart Association functional class III-IV 55%, concomitant coronary artery bypass grafting 41%. RESULTS: We found that 83.7% of patients were in New York Heart Association functional class I and 80.8% had 0 to 1+ aortic insufficiency. Mean gradient at 6 years (n = 75) was 4.4 mm Hg and mean effective orifice area (EOA) 2.4 cm2. Late adverse event rates per patient per year were: embolism 1.0%, endocarditis 0.4%, thrombosis 0%, structural deterioration 0.2%, explant 0.3%, and valve-related death 0.6%. Freedom from valve-related death at 6 years was 95.8%; from cardiac death 96.3%. Freedom from endocarditis was 98.4%, from embolism 93.9%, from structural deterioration 97.4%, and freedom from explant 98.1%. For patients older than 60 years, freedom from structural deterioration was 100%. CONCLUSIONS: These results confirm satisfactory clinical outcomes after aortic valve replacement with the Toronto stentless porcine valve, with a low incidence of valve-related adverse events as long as 96 months after valve replacement. PMID- 11388209 TI - Early and midterm results of model 300 CryoLife O'Brien stentless porcine aortic bioprosthesis. AB - BACKGROUND: The Cryolife O'Brien (CLOB) is a composite stentless bioprosthesis constructed from noncoronary leaflets of three porcine aortic valves. This study aimed to investigate early and midterm results after aortic valve replacement with CLOB xenograft. METHODS: Between 1993 and 2000, the CLOB was implanted in 125 patients (62 men; mean age 71.3+/-6.4 years). Mean prosthesis size was 23.6+/ 2 mm. Mean follow-up time was 37.0+/-12.1 months. Patients underwent echocardiographic studies preoperatively, at discharge, at 6 and 12 months postoperatively, and yearly thereafter. RESULTS: Early (30-day) mortality rate was 2.4% (3 of 125 patients). Of the four late deaths, none was valve related. Actuarial 7-year survival was 93.6%+/-3%. Seven-year freedom from primary valve failure was 98.1%+/-1.8%. All patients showed an improvement of functional status (p < 0.001). ANOVA revealed a significant reduction over time in peak and mean systolic gradients (p < 0.001). Effective orifice area index increased (p < 0.001) and left ventricular mass index significantly reduced in all valve sizes (p < 0.001) during this time interval. CONCLUSIONS: Because the early and midterm results with CLOB xenograft have been satisfactory, we encourage its use as a valve substitute, particularly in patients with small aortic roots. PMID- 11388211 TI - Long-term results after aortic valve replacement with the Biocor PSB stentless xenograft in the elderly. AB - BACKGROUND: This study seeks to define the long-term results after Biocor PSB stentless aortic valve replacement (AVR) in elderly patients, including the effects of No-React treatment. METHODS: We reviewed the outcomes of 106 consecutive patients, aged 70+/-6 years, having Biocor PSB (93 standard, 13 No React) AVR between October 1992 and October 1996. RESULTS: There were three early deaths (3%) and 15 late deaths (15%), during a mean follow-up of 5.8+/-1.6 years. At 8 years, survival was 82%+/-4% and freedom from cardiac death was 94%+/-3%. Freedom from valve failure was 92%+/-4% at 8 years (No-React: 92%+/-8% at 4 years). Replacement of the xenograft was required in 5 patients. Freedom from reoperation was 91%+/-4% at 8 years (No-React: 92%+/-8% at 4 years). Four bleeding and two embolic events were recorded: overall valve-related event-free survival was 81%+/-7% at 8 years (No-React: 76%+/-12% at 4 years). Age of long term survivors averaged 77+/-5 years and their New York Heart Association status was 1.3+/-0.6 (versus 2.9+/-0.6 preoperatively, p = 0.01). CONCLUSIONS: Satisfactory freedom from cardiac events and from valve deterioration added to uniform improvement in functional status despite advanced age and high prevalence of comorbid conditions make AVR with the Biocor PSB xenograft a valid long-term therapy for the elderly. No-React treatment does not influence xenograft durability. PMID- 11388212 TI - Pericardial and porcine stentless aortic valves: are they hemodynamically different? AB - BACKGROUND: We sought to compare the early hemodynamic performance of pericardial stentless aortic valves with that of well-established porcine stentless aortic prostheses. METHODS: A total of 169 patients (97 men and 72 women, aged 73+/-6 years) undergoing aortic valve replacement received either a pericardial (Pericarbon, Sorin Biomedica, Saluggia, Italy; n = 89) or a porcine (Freestyle, Medtronic, n = 80) stentless aortic valve. Aortic valve hemodynamics and root dynamism were assessed by Doppler echocardiography at discharge and 12 months after implantation. RESULTS: Clinical demographic data, valve size (24.0+/-1.9 vs 24.6+/-2.3 mm), and body surface area (1.85+/-0.19 vs 1.80+/-0.19 m2) did not differ between porcine and pericardial valves (both p > 0.05). The 1-year postoperative mean valve pressure gradient (4.2+/-2.6 vs 3.7+/-2.6 mm Hg), effective orifice area (2.2+/-0.8 vs 2.2+/-0.8 cm2), and left ventricular ejection fraction (62+/-13 vs 63+/-13, %) also did not differ (all p > 0.05). However, at discharge, systolic increase in aortic sinus diameter was significantly greater in pericardial valves than in porcine ones (7.7+/-5.7 vs 4.9%+/-4.2%, p < 0.01). Furthermore, pericardial valves had a greater slope of effective orifice area-systolic aortic flow relationship (0.89+/-0.07 vs 0.70+/ 0.06, cm2/100 mL/s, p < 0.01). CONCLUSIONS: Nonprosthetic thin-walled pericardial valves appear to offer better aortic root dynamism and more efficient hemodynamics than those of porcine valves immediately after implant. At 1-year follow-up, however, both types of stentless valves provide equally excellent hemodynamics. The clinical choice between the two will depend on their long-term durability. PMID- 11388213 TI - Hemodynamic evaluation of a new stentless autologous pericardial mitral valve. AB - BACKGROUND: There is no satisfactory mitral valve prosthesis. An ideal mitral valve substitute should be biologic, nonantigenic, and anatomically correct. METHODS: We developed a stentless, chordal-supported (including anterior basal stay chords) mitral valve made with glutaraldehyde-treated autologous pericardium. Eight such prostheses were implanted in sheep. RESULTS: Seven animals survived the operation and were studied postoperatively immediately, at 1 week, and at 1 month. Simultaneous left ventricular and left atrial pressures showed peak and mean transvalvular pressure gradients of 6+/-2 mm Hg and 1+/-1 mm Hg, respectively. Echocardiography performed intraoperatively and then 1 week and 1 month postoperatively showed normal valve leaflet movements. Color and pulsed Doppler echocardiography showed no sign of transvalvular stenosis or regurgitation. Effective orifice area was 5.39+/-0.35 cm2 at intraoperative, 5.51+/-0.29 cm2 1 week after operation (n = 5), and 5.51+/-.28 cm2 1 month after operation (n = 3). Three animals were sacrificed at 19 days and at 1 and 3 months. One animal is alive at 10 months. CONCLUSIONS: This new stentless pericardial mitral valve performed satisfactorily with low gradients and no regurgitation. Possible advantages of this pericardial valve are excellent hemodynamics, ease of construction and implantation, lack of immunogenicity, and low cost. Similarly designed valves but with a shorter nonglutaraldehyde treatment time have been used in 3 sheep monitored for more than 3 months. PMID- 11388214 TI - A new aortic root prosthesis with compliant sinuses for valve-sparing operations. AB - BACKGROUND: We designed and tested a novel aortic root prosthesis with compliant sinuses for valve-sparing operations. METHODS: In eight human aortic roots, the aorta was trimmed 2 mm above the leaflet attachment. The aortic portion of the graft was made by scalloping the Dacron tube. Three sinuses were made individually after turning z-folds in the fabric 90 degrees. Three rectangular pieces were cut and purse strings sewn in each to form the sinuses. The graft was sutured to the aortic root and studied in a left heart simulator. The leaflet motion was recorded (500 frames/second), commissural movement was measured with ultrasound, and the shape of the root was determined from a mold. Seven intact aortic roots were also studied. RESULTS: In the aortic graft roots, the valves were competent and leaflets opened rapidly into a circular orifice, not touching the sinus wall. Commissural diameter increased by 22% when pressure increased from 0 to 80 mm Hg, and increased by a further 6.6% when pressure increased to 120 mm Hg. The sinuses had a teardrop shape. CONCLUSIONS: The dynamics of the aortic graft root and the leaflets were comparable to that of the intact aortic root. This prosthesis is being introduced in clinical practice. PMID- 11388215 TI - Early experience with a quadrileaflet stentless mitral valve. AB - BACKGROUND: Presently no ideal prosthesis for mitral valve replacement exists. The quadrileaflet mitral valve (SJM-Quattro-MV; St. Jude Medical, Inc, St. Paul, MN) is a chordally supported stentless bioprosthesis. Due to its specific geometry it seems to be particularly suited for mitral valve replacement. METHODS: From March 1999 to October 2000, 12 patients (ages 71+/-2 years) received the SJM-Quattro-MV. Six patients suffered from valvular stenosis and 6 patients from incompetence. Preoperatively, all patients were in New York Heart Association functional class III, with left ventricular ejection fraction amounts of 54%+/-17%. RESULTS: Eleven patients received a medium size SJM-Quattro-MV and one patient received a large size SJM-Quattro-MV. Cross-clamp time was 99.8+/-4.9 minutes. Additional procedures were coronary artery bypass grafting (n = 3) and left atrial microwave ablation (n = 2). Postoperative mortality (n = 1) was procedure related. At follow-up of 11.6+/-5.4 months, all patients were well, the transvalvular pressure gradient was 5.0+/-1.4 mm Hg, and the effective orifice area 2.7+/-0.2 cm2. CONCLUSIONS: Our preliminary experiences with the SJM-Quattro MV presented good clinical results and promoted an optimistic way of thinking about the further development of these valve prostheses. PMID- 11388216 TI - Clinical autologous in vitro endothelialization of 153 infrainguinal ePTFE grafts. AB - BACKGROUND: Over the past 17 years, our group has developed and clinically applied an in vitro endothelialization procedure whereby infrainguinal expanded polytetrafluoroethylene (ePTFE) prostheses are confluently lined with cultured autologous endothelial cells before implantation. After a successful randomized pilot study from 1989 to 1993, the procedure was adopted for routine operations. METHODS: Since June 1993, 153 endothelialized ePTFE grafts were implanted in the infrainguinal position in 136 patients (102 above knee (AK) and 51 below knee (BK), 89 men and 47 women, mean age 64.7+/-9.4 years). Seventeen patients received an endothelialized prosthesis bilaterally. Autologous endothelial cells were harvested from 4- to 5-cm segments of a subcutaneous vein (in 86% the cephalic vein), grown to first-passage mass cultures and confluently lined onto 6 (n = 113) or 7-mm (n = 40) inner diameter (ID) ePTFE grafts, precoated with fibrin glue. The observation period for 6-mm grafts was 7 years, and for 7-mm grafts was 4 years. Patency assessment for Kaplan-Meier survivorship analyses was based on duplex sonography and angiography. RESULTS: Kaplan-Meier survivorship function revealed a primary patency rate of 62.8% after 7 years (SE = 0.05) for all infrainguinal reconstructions (60% AK/70.8% BK). The primary patency for stage II and III patients was 64.4% after 7 years. The more recent group of 7-mm ID grafts showed a primary patency of 83.7% after 4 years. CONCLUSIONS: Our data provide strong evidence that autologous endothelial cell lining distinctly improves the patency of small diameter vascular grafts. PMID- 11388218 TI - Midterm results of the Ross procedure. AB - BACKGROUND: The optimal hemodynamic performance and potential growth of the pulmonary autograft has led to expanded indications for the Ross procedure. We reviewed our institutional experience to assess midterm results with the Ross operation. METHODS: In a 7-year period (1992 to 1999), 111 patients with a median age of 15.7 years (range 2 days to 67 years), underwent the Ross procedure. Ninety-five patients had isolated aortic valve disease and 16 pediatric patients had a more complex left ventricular outflow tract obstruction. RESULTS: There were 3 early (2.7%) and 3 late deaths over a median follow-up of 3.6 years (range 6 months to 7.6 years). Actuarial survival at 5 years was 94%+/-2%. In pediatric patients, the pulmonary autograft annulus enlarged from 14.7+/-6.2 mm to 22+/-6.3 mm. This growth followed the expected increase in pulmonary valve diameter based on body surface area. Eight reoperations were necessary for autograft insufficiency at a median interval of 14 months (range 2 days to 31 months). Freedom from replacement of the pulmonary autograft was 91%+/-3% at 5 years. Three patients developed important obstruction of the pulmonary homograft requiring reoperation at a median of 29 months (range 9 to 31 months). CONCLUSIONS: The Ross procedure can be performed with good midterm results. In pediatric patients, autograft growth has been appropriate. The potential for development of important autograft insufficiency suggests close follow-up through the intermediate and late term. PMID- 11388217 TI - Highlights of a ten-year experience with the Ross procedure. AB - BACKGROUND: A review of a 10-year experience with the Ross procedure as a root replacement by a single group of surgeons featuring specific highlights is presented. Highlights include our results from a subset of patients with endocarditis and their management and a comparison of outcomes in patients with aortic insufficiency based on technical changes made after 5 years' experience. METHODS: The total patient group was 191, with 148 male and 43 female participants with an age range from 1 day to 69 years. Five of the patients in the 0-to-20 age group were newborns. Fifty-three of the adults were operated on for infectious endocarditis. In the total patient group 43% had aortic insufficiency, 28% had aortic stenosis, and 29% had mixed disease. RESULTS: Operative mortality was 5.2% with a late mortality of 2.6%. The actuarial survival was 90.2% at 10 years. Freedom from autograft explantation was 93.2% and freedom from homograft replacement was 98.4%. The endocarditis patients had an operative mortality of 3.8% with 100% cure of the infection. Freedom from reinfection on the autograft was 98.1%, and freedom from infection of the pulmonary homograft was 98.1%. The actuarial survival was 86.3%. CONCLUSIONS: A specific review of the patients with aortic insufficiency resulted in a failure of the autograft in 7 patients among a cohort of 41 during the first 5 years of the study. After a change in technique in which the aortic annulus is narrowed and fixed to a measured size to match the body surface area, we have had no failures in the autograft. Although these results are early, we believe that these data support the use of the autograft as an excellent choice for replacement of the aortic valve in infective endocarditis. Finally, the use of the autograft for aortic insufficiency is reasonable with fixation of the aortic annulus so that subsequent dilation does not occur. PMID- 11388219 TI - Technical evolution of the Ross operation: midterm results in 186 patients. AB - BACKGROUND: The Ross operation approaches the ideal aortic valve replacement. Between February 1995 and February 2000 we performed 186 procedures. This article reviews modifications introduced reflecting our experience. METHODS: In all patients the Ross operation was performed as root replacement. Echocardiographic follow-up was complete in 94% of patients. RESULTS: No operative death or early mortality occurred, nor did thromboembolic or hemorrhagic events. One patient died at 25 months from hemoptysis with pulmonary valve vegetations. Three patients required reoperation for autograft insufficiency. In 1 patient a tethered cusp was repairable and in 2 patients progressive autograft dilatation required autograft replacement. After routinely incorporating support into the aortic annulus and replacing all dilated ascending aorta, autograft dilatation did not recur. For the pulmonary homograft, one outflow patch was placed to relieve a symptomatic gradient. Nine patients with elevated gradients were under observation. Echocardiography revealed autograft median peak systolic gradients of 4.6+/-2.8 mm Hg, pulmonary homograft gradients of 14.8+/-9.6 mm Hg, and nil or insignificant regurgitation. CONCLUSIONS: The aortic annulus must be supported and the dilated ascending aorta replaced. Root replacement with a short autograft allows consistent results. Pulmonary homograft dysfunction is rare but unpredictable. PMID- 11388220 TI - Estimated event-free life expectancy after autograft aortic root replacement in adults. AB - BACKGROUND: Autograft aortic root replacement is an established therapeutic option for young adults with aortic valve disease. Unfortunately, most series are small with a limited follow-up. Meta-analysis and microsimulation modeling were used to predict long-term outcome based on currently available midterm data. METHODS: We combined our center's experience with autograft aortic root replacement in 85 adult patients in a meta-analysis with reported results of three other hospitals. The outcomes of this meta-analysis were entered in a microsimulation model, calculating (event-free) life expectancy after autograft aortic root replacement. RESULTS: The pooled results comprised 380 patients with a total follow-up of 1,077 patient-years. Mean age was 37 years (range 16 to 68 years). Male/female ratio was 2.7. Operative mortality was 2.6% (n = 10); during follow-up 6 more patients died. Linearized annual risk estimates were 0.5% for thromboembolism, 0.3% for endocarditis, and 0.4% for nonstructural valve failure. Structural autograft failure requiring reoperation occurred in 5 patients, and a Weibull function was constructed accordingly. Using this information, the microsimulation model predicted age- and gender-specific mean, reoperation-free, and event-free life expectancy. CONCLUSIONS: Based on current evidence the calculated average autograft-related reoperation-free life expectancy is 16 years. The combination of meta-analysis and microsimulation provides a promising and powerful tool for estimating long-term outcome after aortic valve replacement. PMID- 11388221 TI - Hemograft crossmatching is unnecessary due to the absence of blood group antigens. AB - BACKGROUND: Homograft valves are subject to calcification and structural degeneration in the long term. Blood group matching is performed in many centers, and it remains controversial whether immunologic responses associated with potential blood group incompatibility contribute to the degeneration of unmatched homografts. We studied the expression of carbohydrate blood group antigens on valve endothelium of thawed aortic homograft valves and freshly harvested human cardiac valves. METHODS: Cryopreserved human aortic homograft valves and freshly harvested human aortic, pulmonary, mitral, and tricuspid valves were incubated with antibodies to A, B, and O blood group antigens. RESULTS: Cardiac microvascular endothelium stained positively with antiendothelial CD31 antibody in both cryopreserved and fresh tissue. Cryopreserved valve endothelial lining rarely stained positively for CD31, in contrast to fresh valves, which always stained positive. Cryopreserved or fresh cardiac microvascular endothelium strongly expressed A, B, or H antigens. In contrast, ABH antigens were not detectable on homograft or fresh cardiac valve endothelium. CONCLUSIONS: The absence of expression of carbohydrate antigen on valvular endothelium suggests that blood group incompatibility does not play a significant role in homograft degeneration. PMID- 11388222 TI - Results of up to 9 years of high-temperature-fixed valvular bioprostheses in a young population. AB - BACKGROUND: Bioprosthetic valve replacement in young patients remains a controversial issue due to a high rate of early calcification. Previous studies in our laboratory have shown that high-temperature fixation of glutaraldehyde preserved bioprosthesis (HTF) mitigates calcification. The first clinical application of this technique was started in 1991. METHODS: From January 1991 to September 1998, 50 patients in whom anticoagulants were contraindicated underwent single aortic valve replacement (n = 33) or mitral valve replacement (n = 17) using HTF bioprostheses. The age of the patients ranged from 7 months to 35 years (mean 22.7+/-6.8 years). The mean New York Heart Association status was 2.4. Mean follow-up 4 years +/- 1.8 for a total follow-up of 196 patient-years. RESULTS: There were no operative deaths and but there were two late deaths, one valve related. Structural failure occured in 4 patients (2%/patient-year) requiring a reoperation in 3 patients (1.5%/patient-year). No endocarditis or thromboembolic episodes were observed. At late examination (June 2000), 46 patients (92%) were in New York Heart Association class I or II, with a well functioning valve. CONCLUSIONS: Replacement with HTF bioprostheses in young patients has demonstrated encouraging midterm results with a low incidence of structural failure PMID- 11388223 TI - Biological versus mechanical aortic valve replacement in children. AB - BACKGROUND: Aortic valve replacement in children remains challenging because of constraints imposed by available prosthetic devices. Potential risks of anticoagulation with mechanical valves and degeneration of other biological substitutes have kindled interest in the Ross procedure. This study outlines the evolution of our 27-year experience with prosthetic devices. METHODS: Ninety-nine patients who underwent aortic valve replacement (January 1973 through September 2000) were included in this study. Procedures included implantation of pulmonary autograft (PA) (n = 42), aortic homograft (AH) (n = 3), mechanical valves (MV) (n = 41), and xenograft tissue valves (XG) (n = 13). RESULTS: The mean follow-up times were: 3.8+/-1.3 years for PA, 3.5+/-1.5 years for AH, 7.7+/-4.7 years for MV, and 8.4+/-4.8 years for XG. There were no significant differences in perioperative outcomes among the groups (p < or = 0.05) or early deaths (2 each in the MV, AH, and PA groups). The incidence of valve-related complications and reoperations was high in the MV (n = 5), XG (n = 7), and AH (n = 1) groups as compared with the PA group (n = 3, p < 0.01). Early and late mortality for the series was 8.6% (n = 8). Overall, the reoperation rate was 20.7% (n = 18): 15.2% (5 of 33) MV, 70% (7 of 10) XG, 50% (1 of 2) AH, and 11.9% (5 of 42) for PA. The actuarial survival rate was 87.8% and 100% at 10 years for MV and XG, and 95.2% and 6.6% at 7 years for PA and AH. CONCLUSIONS: Aortic valve replacement in children can be performed with acceptable mortality and good long-term results. The Ross procedure, although more complicated, has the advantage of not requiring anticoagulation therapy, can be performed in all age groups, possesses inherent growth potential, and exhibits the most normal left ventricular outflow tract hemodynamics. PMID- 11388224 TI - Use of the Medtronic Freestyle valve as a right ventricular to pulmonary artery conduit. AB - BACKGROUND: We have used the Medtronic Freestyle bioprosthesis as a right ventricular to pulmonary artery conduit recently in an attempt to overcome some of the problems associated with homografts and stented xenografts. The aim of this study was to review the performance of this prosthesis. METHODS: Prospectively collected data for patients having Freestyle bioprostheses implanted as a right ventricular to pulmonary artery conduit were reviewed to assess clinical outcome and echocardiographic results. RESULTS: Thirteen patients aged 13 days to 22.5 years (median, 7.9 years) underwent either primary repair (n = 5) or change of conduit (n = 8) using the Freestyle bioprosthesis. One neonate with truncus arteriosus died postoperatively of pulmonary hypertension. One conduit was explanted 27 months after repair of neonatal truncus arteriosus. There has been no incidence of significant prosthetic regurgitation, thromboembolism, or endocarditis at mean follow-up of 10.1 months (range, 2 weeks to 29 months). CONCLUSIONS: The Medtronic Freestyle valve is a reliable pulmonary valve substitute in the short term. Early results justify continued clinical use of the device in this setting with close follow-up. PMID- 11388226 TI - Electron beam tomography for cusp calcification in homograft versus Freestyle xenografts. AB - BACKGROUND: We have previously shown, by means of electron beam tomography, the pattern of calcification of the aortic root wall of homografts and porcine xenografts after aortic root replacement. However, application of similar methods for cusp calcification raises specific problems that have not been addressed before. METHODS: A new method for localizing and quantifying calcification of the aortic valve cusps has been evolved. Intravenous contrast-enhanced electron beam tomography was introduced to visualize the aortic cusps. This technique was applied to quantify cusp calcification in 37 patients after aortic root replacement with a homograft (group H) or a Medtronic Freestyle valve (group F) at set intervals between 6 months and 2 years. A calcification score in Hounsfield units (HU) and a calcified volume score in cubic millimeters were calculated. RESULTS: The aortic leaflets were clearly visualized in all patients. The mean calcium score in the cusps was 28.8+/-64.4 HU in group F and 62.4+/-66.9 HU in group H (p = not significant). The mean calcified volume score was 327.0+/ 425.9 mm3 in group F and 642.0+/-443.0 mm3 in group H (p = not significant). CONCLUSIONS: Contrast enhancement electron beam tomography is a useful tool for quantification of calcium in the aortic valve leaflets. Our preliminary results show a tendency toward more calcification in the homografts. This needs to be studied further in a bigger cohort of patients followed up for longer periods. PMID- 11388225 TI - Performance of allografts and xenografts for right ventricular outflow tract reconstruction. AB - BACKGROUND: We compared the long-term durability of allografts and xenografts implanted for reconstruction of the right ventricular outflow tract. METHODS: A total of 401 patients were studied from January 1974 to June 2000 (145 xeno- and 256 allografts), follow-up being 98% complete. We analyzed freedom from reoperation and allograft specific factors that may indicate degeneration. RESULTS: The age at implantation was 2 days to 31 years (median 4.0 years). Conduit exchange rate was similar (p = 0.2) for conduit diameters less than 15 mm (41%+/-9% for allografts, 30%+/-6% for xenografts), but significantly different (p = 0.02) for diameters of 15 mm or larger (60%+/-8% for allografts, 30%+/-10% for xenografts). Diagnosis-related 20-year survival analysis showed a significantly (p = 0.01) better survival of patients with tetralogy of Fallot/pulmonary atresia (83%+/-5%) and Rastelli-type surgery (81%+/-8%) compared with patients with truncus arteriosus communis (69%+/-8%). ABO-compatibility, preservation method, and aortic or pulmonary allograft could not be identified as risk factors for allograft longevity. CONCLUSIONS: For smaller diameters (less than 15 mm), allografts exhibit no advantage over xenografts, whereas in larger diameters (15 mm or larger) allografts are the conduit of choice for the right ventricular outflow tract. PMID- 11388227 TI - Repair of truncus arteriosus in early infancy with antibiotic sterilized aortic homografts. AB - BACKGROUND: To evaluate the fate of the truncal valve, the antibiotic sterilized aortic homografts, and the survival after repair of truncus arteriosus in the first 6 months of life. METHODS: Between 1974 and 1994, 23 infants (mean age 1.7 months, range 5 days to 6 months) underwent primary repair of truncus arteriosus by one surgeon (J.L.M). Sixteen were neonates (age range 5 to 30 days). Continuity between the right ventricle and the pulmonary artery was established with an aortic antibiotic sterilized homograft (mean diameter 14.9 mm, range 11 to 17 mm). Follow-up was 100% complete. RESULTS: Four neonates with severe truncal regurgitation died early (17.4%). Fourteen patients underwent reoperations. Five had a truncal valve replacement (mean time 7 years, range 6 months to 17 years). Ten-year freedom from truncal valve replacement was 78.2%. Eleven patients had homograft replacement (mean time 12.7 years, range 2 to 26.2 years). Ten-year freedom from homograft replacement for any cause was 77.1% (for homograft-related problems it was 86.7%). Seven patients retained the original homografts (mean time 14.3 years, range 6 to 18.7 years). There was one late death. Overall 10-year survival was 79% and for the hospital survivors it was 95%. All survivors are in New York Heart Association functional class I. CONCLUSIONS: Abnormal truncal valves pose serious early and late problems but the patients with normal truncal valves do well and seem unlikely to need replacement of these valves. The durability of the antibiotic sterilized aortic homograft even in sizes less than 14 mm is remarkably good. Late survival is excellent. PMID- 11388228 TI - Effect of altered hydration on the internal shear properties of porcine aortic valve cusps. AB - BACKGROUND: Dehydration of tissue due to glutaraldehyde fixation has been reported and was examined in this study of porcine aortic valve cusps. The effect of altered hydration on cusp internal shear properties was also examined. METHODS: Hydration level was assessed by wet mass measurement of cusps stored in solutions for times up to 1000 minutes. Solutions used in this study included Hanks solution, porcine blood, 0.5% glutaraldehyde, and several dextran solutions. Shear testing was performed on physiologically hydrated, superhydrated, and dehydrated cusps. RESULTS: There was very little difference between the physiologic and superhydrated leaflets; however, dehydration caused significant stiffening with increased hysteresis and stress relaxation. CONCLUSIONS: Glutaraldehyde has been shown to increase shear stiffness of valve cusps. Tissue dehydration also increased shear stiffness but increased stress relaxation and hysteresis, which was contrary to observations reported after glutaraldehyde fixation. The significant effect of dehydration on cusp mechanical properties does not account for the effects observed after glutaraldehyde fixation, but it demonstrates that hydration level is an important factor that affects internal shear properties of valve cusps. PMID- 11388229 TI - Immunological aspects of fresh and cryopreserved aortic valve transplantation in rats. AB - BACKGROUND: The influence of immune activation on valve allograft degeneration remains unclear. We studied the combined effect of major histocompatibility complex (MHC)-incompatibility and cryopreservation on valve performance, histomorphology, and tissue antigenicity in rats. METHODS: Fresh or cryopreserved allogeneic aortic valves from WAG (RT1u) rats were transplanted to DA (RT1a) recipients and syngenic transplants served as controls. After 7 or 21 days, valves were examined for competence and morphology. Immune reactivity of the recipient was measured by concanavalin A (conA) stimulation and analysis of donor reactive Helper T-lymphocyte frequencies (HTLf) in peripheral blood and spleen. RESULTS: Syngenic grafts demonstrated normal competence and structure. Allografts lost their competence over time caused by destruction of the leaflets combined with cellular infiltration in the vascular wall. Cryopreservation induces early loss of competence and retrovalvular thrombosis. Cryopreserved allografts were also heavily infiltrated. ConA stimulation indices and HTLf were higher in allogeneic recipients compared to syngenic recipients (p < 0.03). Cryopreserved allografts elicited a lower immune response compared with fresh allografts (p < 0.03). CONCLUSIONS: Aortic valve allografts are able to induce a donor-reactive immune response that is related to early graft destruction and incompetence. Cryopreservation appears to diminish but not eliminate the antigenicity of the allograft. PMID- 11388230 TI - Characterization of the immune response to valve bioprostheses and its role in primary tissue failure. AB - BACKGROUND: The role of an immune response in the failure of bioprosthetic heart valves is poorly understood and disregarded by many. To elucidate the nature of the immune response to glutaraldehyde-treated tissue and the possible role of graft-specific antibody in graft mineralization, we performed immune calcification studies in the rabbit and correlated those results with the analysis of specific antibodies. METHODS: Aortic wall buttons (6 mm) were punched from porcine aortic wall tissue fixed with 0.2% glutaraldehyde and detoxified with urazole and then subsequently perforated under sterile conditions. The perforated buttons were then incubated with either immune serum prepared by immunization of New Zealand White rabbits (n = 5) with Freund's incomplete adjuvant emulsions of tissue homogenates of similarly treated aortic wall tissue, or incubated with the corresponding control preimmune sera obtained before immunization of the same animals. The tissue was then implanted subdermally on the back of unrelated New Zealand White rabbits (n = 8) for a period of 3 weeks. After the buttons were explanted, tissue calcium levels were determined by atomic absorption spectroscopy. RESULTS: Tissue calcium was increased in all five immune serum-treated replicates (range, 61.8% to 431.2%; mean, 225.9%+/-73.2%) when compared with control samples treated with preimmune sera. Overall, the mean calcium level was significantly increased (p < 0.0001) when tissue was treated with immune sera (66.0+/-10.0 microg/mg versus 22.6+/-4.8 microg/mg in control tissue). Graft specificity of immune sera was confirmed by Western blot analysis. CONCLUSIONS: These results strongly suggest a role of circulating graft-specific antibody in the disease of bioprosthetic graft calcification. PMID- 11388231 TI - Transgenic porcine valves show no signs of delayed cardiac xenograft rejection. AB - BACKGROUND: Glutaraldehyde fixation stiffens the structural integrity of porcine valves, although the solution also destroys tissue viability and accelerates calcification. Recently, we demonstrated that fresh cardiac valves from domestic pigs do not express the galactose alpha1, 3 galactose (alpha-Gal) antigen and may be immunologically unique. The absence of alpha-Gal explained why the valves remained pristine while the rest of the porcine heart was destroyed by primate immunoglobulin M (IgM) and complement membrane attack complex (MAC) within 60 minutes. We sought to clarify whether fresh porcine valves from transgenic pigs bearing human complement regulatory proteins (CD59/DAF) can survive longer in primates and whether porcine cardiac valves remained immunologically privileged after prolonged exposure. METHODS: Tissue sections from wild-type untransplanted (n = 6), wild-type transplanted (n = 3), and transgenic pigs expressing human CD59/DAF proteins transplanted (n = 3) porcine-to-primate cardiac grafts were examined by hematoxylin and eosin, and by immunohistochemistry for the porcine endothelial marker (GalNac), alpha-Gal, primate IgM and MAC. RESULTS: alpha-Gal antigens were highly expressed on the vascular, but not valvular, endothelium of transgenic pigs. Hearts from CD59/DAF transgenic pigs survived 5, 7, and 11 days, but showed increasing IgM and MAC deposition until failure. Valves remained morphologically intact at explant, and strong GalNac staining suggested an intact endothelial surface. However, the valves showed no signs of IgM- or MAC-mediated damage. CONCLUSIONS: Although hearts from transgenic pigs expressing human complement regulatory proteins can survive for days in the primate recipient, the xenografts eventually fail because of escalating attacks of primate IgM and MAC. The absence of the alpha-Gal antigens protects unfixed porcine valves from rejection. PMID- 11388232 TI - Decreased porcine valve antigenicity with in vitro culture. AB - BACKGROUND: Porcine valvular prostheses may stimulate inflammation after implantation, with resultant accelerated structural degeneration. We investigated the expression of porcine major histocompatibility complex (MHC) class II molecules on valve leaflets and the possibility of decreasing valve antigenicity with in vitro culture. METHODS: Aortic and pulmonary valves were harvested from domestic pigs under sterile conditions and cultured in vitro with either porcine or baboon serum for 4 days. Valves were harvested daily and fixed in Carnoy's or formalin solution. Microtome sections of valves were examined by hematoxylin and eosin, and by immunohistochemistry for porcine MHC class II proteins and an endothelial marker, alpha-N-acetylgalactosaminyl glycoprotein (alpha-GalNac). RESULTS: Porcine aortic and pulmonary valves constitutively express alpha-GalNac proteins and porcine MHC class II antigens. Porcine valves continue to express both alpha-GalNac and MHC class II after 48 hours of culture in porcine serum. After 48-hour culture in baboon serum, however, MHC class II antigens became undetectable on valvular leaflets, although alpha-GalNac molecules were still detected. CONCLUSIONS: Porcine valvular endothelial cells remain viable after 2 days of in vitro culture. Porcine valves cultured with primate serum show decreased MHC class II antigenic expression. In vitro culture before glutaraldehyde fixation may decrease inflammation associated with implantation. PMID- 11388233 TI - Effect of human immunoglobulins on the immunogenicity of porcine bioprostheses. AB - BACKGROUND: Glutaraldehyde fixation (GT) is known to reduce immunologic reactions and tissue degeneration after implantation in humans. Sterilization after glutaraldehyde fixation (G-ST) improves the safety and reduces the tendency of GT valves to calcify. Intravenous immunoglobulins (IVIg) have been shown to reduce xenogeneic response against porcine tissue. We have investigated the effect of these fixation procedures combined with and without IVIg on the antigenicity of bioprostheses. METHODS: Lewis adult rats were implanted subcutaneously with a fresh, GT, or G-ST porcine heart valve pre- or posttreated with different amounts of IVIg. We followed by enzyme-linked immunosorbent assay and IgM and IgG titers against protein extracts from the porcine heart valves after implantation. Cellular reactivity was assessed in xenogeneic lymphoendothelial coculture experiments. Calcification content was also examined. RESULTS: Glutaraldehyde fixation partially decreased the humoral response against proteins of the implant but elicited a cellular xenogeneic response. Sterilization reduced these reactivities, but retained antigenicity. Intravenous immunoglobulin postincubated with GT valves before implantation reduced the antigenicity of the tissue to the same extent as G-ST valves, but had no effect on valvular tissue calcification. CONCLUSIONS: Our studies demonstrate that IVIg or the sterilization procedure (ST) reduced the cellular response against glutaraldehyde-fixed valves (GT), whereas reduced calcification was observed only with ST. PMID- 11388234 TI - Calcification characteristics of porcine stented valves in a juvenile sheep model. AB - BACKGROUND: Different antimineralization treatments of stented porcine bioprostheses were evaluated: ethanol (Epic), alpha-amino-oleic acid (AOA) (Mosaic), and sodium dodecyl sulfate (SDS) (Hancock II). A nontreated, glutaraldehyde-fixed valve (Labcor) served as control. METHODS: For each treatment, six valves were implanted in juvenile sheep in the pulmonary position. Valves were explanted after 3 and 6 months and examined macroscopically, by roentgenogram and light and transmission electron microscopy. Calcium content (microg/mg) was determined by atomic absorption spectrometry. RESULTS: The Labcor valves revealed small calcium deposits in the cusps, although calcium content remained low (median value 0.4+/-0.8 microg/mg). SDS did not prevent cusp calcification as assessed by histology and calcium content measurement, which was higher than in all other valves: 1.9+/-4.6 microg/mg (p < 0.05). Cusp retraction and rupture were occasionally found in the Hancock. The Mosaic and Epic valves showed no cusp calcification and had low calcium contents (0.3+/-2.4 microg/mg and 0.7+/-0.6 microg/mg, respectively). Epic showed less pannus formation, but had hematoma or iron staining in the cusps. CONCLUSIONS: SDS is inefficient as an antimineralization treatment, in contrast to ethanol or AOA. Cusp hematoma after ethanol treatment needs further investigation. PMID- 11388235 TI - Protein adsorption of calcified and noncalcified valvular bioprostheses after human implantation. AB - BACKGROUND: The incidence of calcification of porcine valve bioprostheses shows important, and as yet unexplained, variations. Previous studies by others showed that osteopontin and osteocalcin are expressed in calcified porcine valve bioprostheses. However, no study has yet explored other proteins that could also be involved. METHODS: Twelve porcine valve bioprostheses were retrieved from 12 patients and were separated into two groups. Group 1 (n = 6) had early calcification after 4 to 9 years (mean, 6+/-2.3 years). The mean age of the patients at the time of implantation was 46+/-9 years. Group 2 (n = 6) had no calcification after 8 to 14 years (mean, 12+/-2.8 years). The mean age was 47+/ 13.4 years. These valves were analyzed by electrophoresis, and the bands were quantified by densitometry. RESULTS: A 14-kd protein showed a 50% increase in the calcified group. A 31-kd protein found in the calcified group was not detected in the noncalcified group. Three other proteins (45, 39, and 28 kd) showed reduced adsorption in the calcified group. CONCLUSIONS: Important differences were found in the proteins adsorbed in calcified and noncalcified bioprostheses after implantation in patients. Besides osteopontin and osteocalcin, several other proteins may play a role in the process of calcification of valvular bioprostheses. PMID- 11388236 TI - Protein adsorption in glutaraldehyde-preserved bovine pericardium and porcine valve tissues. AB - BACKGROUND: Proteins adsorbed by bioprosthetic tissues after implantation play a major role in the process of calcification. We investigated whether there are differences in protein adsorption between bovine pericardial and porcine valvular tissues that could correlate with the differences observed clinically. METHODS: Glutaraldehyde-treated bovine pericardial and porcine valve samples were implanted subcutaneously in rats and retrieved 1 month after implantation. Total protein content was assessed by Lowry's method. Qualitative analysis was performed by polyacrylamide gel electrophoresis. Quantitative analysis was performed by densitometry. RESULTS: Total protein content showed a higher protein concentration in porcine valve tissue than in pericardial tissue: 149+/-22.6 microg/mg dry tissue versus 108+/-12.7 microg/mg dry tissue (38% increase). In pericardial tissue, four protein bands (17, 16, 15.5, and 13.5 kd) showed decreased concentration when compared with porcine valve tissue, whereas one band (11 kd) showed increased concentration. CONCLUSIONS: Significant differences were found in protein content between bovine pericardial and porcine valve tissues. Correlations with clinical findings may lead to a better understanding of the mechanism involved in the process of calcification, particularly the role played by the structure of the tissues. PMID- 11388237 TI - Biochemical properties of heat-treated valvular bioprostheses. AB - BACKGROUND: Preliminary studies showed that heat treatment of glutaraldehyde preserved valvular bioprostheses mitigates calcification. This study was carried out to define the physicochemical characteristics of the heat-treated tissues to elucidate the mechanism involved in the mitigation. METHODS: Glut bovine pericardium or porcine valve samples were treated at 50 degrees C in a 0.625% glutaraldehyde solution for 2 months. Some samples underwent assay for shrinkage temperature, moisture content, ninhydrin test, and acid hydrolysis, and other samples were incubated in human serum for 3 days and then analyzed by electrophoresis to study protein adsorption. RESULTS: Heat treatment mitigated calcification without adversely affecting shrinkage temperature (84.81 degrees C versus 83.95 degrees C) and moisture content (78.68% versus 78.71%). A significant reduction in free amino groups (0.15 versus 0.37 mol NH2/mol collagen) and a significant increase in resistance to acid hydrolysis were observed. Total protein content was similar, but significant differences were found for four proteins adsorbed in the tissues (167, 45, 11.6, and 10 kDa). CONCLUSIONS: The anticalcification effect of heat treatment may be attributed to structural changes, lipid extraction, increased resistance, and modifications of the type and concentration of the proteins adsorbed in the tissue. PMID- 11388238 TI - Effect of ethanol and ether in the prevention of calcification of bioprostheses. AB - BACKGROUND: Lipids play a significant role in the process of calcification of bioprostheses. We assessed whether lipid extraction by ethanol, ether, or a surfactant could mitigate calcification of glutaraldehyde-treated bioprostheses. METHODS: On 200 bovine pericardium samples pretreated with 0.6% glutaraldehyde, lipid extraction was carried out by ethanol, ether, or the tween 80 surfactant, and combinations thereof. The treated tissues were implanted subcutaneously in 50 juvenile rats for 4 and 6 months. Lipids were analyzed by Fourier transform infrared spectrophotometer and chromatography before implantation. Calcium content of implanted tissues was assessed by atomic absorption spectrometer. RESULTS: Ethanol, ether, or surfactant did mitigate calcification. The most efficient pretreatments were the combination of ethanol and surfactant (calcium content: 15.5+/-6.8 microg/mg dry tissue after 6 months implantation) or the combination of ethanol, ether, and surfactant (13.1+/-6.2 microg/mg dry tissue) when compared with surfactant alone (42.9+/-12.7 microg/mg dry tissue). CONCLUSIONS: Ethanol or the combination of ethanol and ether added to the currently used glutaraldehyde-surfactant treatment further mitigates calcification. PMID- 11388239 TI - Tissue characterization and calcification potential of commercial bioprosthetic heart valves. AB - BACKGROUND: Tissue properties may contribute to intrinsic calcification of bioprosthetic heart valves. Phospholipids have been proposed as potential nucleation sites for calcification. Other tissue properties might also be important in calcification. METHODS: Commercial and control bioprosthetic valve tissues were characterized by shrinkage temperature, moisture content, free amine content, phospholipid content, and calcification level after 90-day rat subcutaneous implantation as described. RESULTS: Shrinkage temperature, moisture content, and free amine content were typical for glutaraldehyde-cross-linked tissues. Phospholipid and calcium levels varied considerably among valve types. There was a significant correlation between phospholipid levels and calcification (r = 0.63, p = 0.04). Sulzer Carbomedics Mitroflow and Toronto SPV valve tissues had significantly more calcification than other commercial bioprostheses in this study (p < 0.01). Carpentier-Edwards Duraflex, CE SAV, and CE PERIMOUNT valve tissues had significantly less calcification than Medtronic Mosaic in this animal model (p < 0.02). CONCLUSIONS: Processes that reduce phospholipid levels are associated with reduced calcification in the rat subcutaneous model. Significant differences in calcification level were found among commercially available valves. The clinical significance of these results is unknown. PMID- 11388240 TI - Stentless bioprosthetic heart valve research: sheep versus primate model. AB - BACKGROUND: The mild inflammatory response against stented bioprosthetic heart valves in the sheep model is often opposed by a more distinct response in failing human implants. With the emergence of stentless root prostheses with their significantly larger proportion of tissue interacting with the immune system of the host, a more relevant animal model than the sheep may be needed. METHODS: Valved, porcine aortic roots of 5 cm length were fixed in 0.2% glutaraldehyde and implanted in the upper descending aorta of Merino sheep (n = 5; 43+/-3 kg) and Chacma baboons (n = 5; 17+/-3 kg). After 6 weeks of tissue calcification, pannus outgrowth and inflammation were assessed by atomic absorption spectrophotometry, histologic damage scoring (0 to 3), image analysis, and transmission electron microscopy. RESULTS: The main difference between the two animal models was in aortic wall calcification (64.8+/-39.8 microg/mg in the sheep model versus 4.1+/ 5.9 microg/mg in the primate model; p > 0.005). In both models, leaflet calcification was negligible (2.6+/-2.4 microg/mg in the sheep versus 2.5+/-1.9 microg/mg in the primate), and the overall extent of inflammation was comparable (1.2+/-0.8 versus 0.98+/-0.7; p = 0.18 in the sheep and the primate, respectively). Qualitatively, the sheep demonstrated a macrophage-dominated reaction whereas the inflammatory demarcation often resembled a granulocyte dominated xenograft response in the primate. Pannus outgrowth was comparable in length (8.4+/-2.3 mm versus 9.1+/-4.3 mm proximally and 7.1+/-3.4 mm versus 7.4+/ 5.1 mm distally, in the sheep and baboon, respectively; p > 0.05). CONCLUSIONS: Our results confirm the sheep as a significantly stronger calcification model for stentless aortic heart valves than the primate. Remaining antigenicity of porcine tissue as a result of incomplete cross-linking, however, elicits a distinctly stronger xenograft-type reaction in the primate model. PMID- 11388241 TI - Decellularized human valve allografts. AB - BACKGROUND: Variable performance of allograft tissues in children and some adults may be linked to an immune response and could be mitigated by reducing implant antigenicity. METHODS: As endothelial and fibroblast cells are the likely source of valve antigenicity, human (CryoValve SG) and sheep pulmonary valves were decellularized using the SynerGraft treatment process. Treated valves were evaluated in vitro using histochemical, biomechanical, and hydrodynamic methods, and compared with standard cryopreserved valves. Four SynerGraft-treated and two cryopreserved sheep pulmonary valves were implanted as root replacements in the right ventricular outflow tract of growing sheep and monitored echocardiographically and histologically at 3 and 6 months. CryoValve SG human pulmonary valves were implanted in 36 patients. RESULTS: SynerGraft treatment reduced tissue antigen expression but did not alter human valve biomechanics or strength. Decellularized sheep allograft valves were functional during the implantation period, and, they became progressively recellularized with recipient cells. In humans, CryoValve SG pulmonary valves did not provoke a panel reactive antibody response. CONCLUSIONS: SynerGraft decellularization leaves the physical properties of valves unaltered and substantially diminishes antigen content. Reduction in implant cellularity enables host recellularization of the matrix, which should favorably impact long-term graft durability. PMID- 11388242 TI - Transformation of nonvascular acellular tissue matrices into durable vascular conduits. AB - BACKGROUND: Prosthetic grafts commonly used for vascular reconstruction are limited to synthetics and cross-linked tissue grafts. Within these devices, graft infections are common, compliance mismatch is significant, and handling qualities are poor. Natural biological tissues that are unfixed have been shown to resist infections and be durable and compliant. A natural biological matrix that could be remodeled appropriately after implantation would be a desirable graft for vascular reconstruction. METHODS: SynerGraft tissue engineering strategies have been used to minimize antigenicity and produce stable unfixed vascular grafts from nonvascular bovine tissues. These grafts have replaced the abdominal aortas of 8 dogs that have been followed for up to 10 months. RESULTS: Early evaluation indicates rapid recellularization by recipient smooth muscle actin positive cells, which become arranged circumferentially, into the media. Arterioles were present in the adventitial areas and endothelial cells were seen to cover lumenal surfaces. After 10 months, grafts were patent and not aneurysmal. CONCLUSIONS: These data indicate that SynerGraft processing of animal tissues is capable of producing stable vascular conduits that exhibit long-term functionality in other species. PMID- 11388243 TI - Surface attached ultrathin polymer monolayers for control of cell adhesion. AB - BACKGROUND: Calcific degeneration is the major drawback of bioprostheses. None of the numerous preventive approaches omitted calcification. Previous studies showed that cellular surface seeding decreases calcium uptake in vitro but achievement of coverage remains problematic. A new approach is presented masking glutaraldehyde residues with a polymer layer allowing cell seeding. The aim of this study was to evaluate different polymers for suitability. METHODS: Ten polymers--covalently bound to glass--were tested for their ability to seed animal and human cells. Quality of coverage was evaluated by light and scanning electron microscopy, and polymers were characterized physicochemically. RESULTS: Quality of cellular growth was similar for canine and human cells. Five polymers allowed excellent surface coverage, two led to a decrease of cell adherence, and four to poor cellular growth. No correlation between molecular weight, thickness, hydrophilicity, or charge of the polymer and cell growth was found. CONCLUSIONS: Polymer monolayers can promote cellular growth but without correlation to physicochemical characteristics. Polymers covalently bound to biologic tissue appear to be a promising approach for achieving cellular coverage of biomaterials. PMID- 11388244 TI - Applicability of deconvolution and nonlinear optimization for reconstructing optical images from near-field optical microscope images. AB - We have made a computer reconstruction of a nanometric optical image of a sample from an observed near-field optical image. The near-field microscope image used for investigation was obtained numerically in three dimensions with the finite difference time-domain (FDTD) method. The sample is dielectric substrate containing nanometric two strips made of dielectric or metal. Deconvolution with non-negativity constraint is used to reconstruct the nanometric structure of dielectric strips, while nonlinear optimization is used to reconstruct the metallic strips. The difference in choice of reconstruction method between two samples comes from the difference in degree of interaction or multiple scattering of the sample structure and the probe. It is shown in the results, the resolution limit attainable in deconvolution is as fine as the size of aperture of probe. PMID- 11388245 TI - 3D simulations of the experimental signal measured in near-field optical microscopy. AB - We present three-dimensional simulations of the image formation process in near field optical microscopy. Our calculations take into account the different components of a realistic experiment: an extended metal coated tip, a subwavelength sample and its substrate. We investigate all possible detection (transmitted, reflected and collected field) and scanning (constant height, constant gap) modes. Our results emphasize the strong influence of the tip motion on the experimental signal. They also show that it is possible, by controlling the polarization of both the illumination and the detected field, to strongly reduce these artefacts. PMID- 11388246 TI - Theoretical comparison of illumination and collection mode images of magneto optical dots. AB - We compare theoretical images of the same sample obtained with two different scanning near-field optical microscopes. The sample is a two-dimensional periodic array of magnetic sub-micrometric dots. The magnetization is perpendicular to the sample plane (polar magnetization). The first configuration is a scanning tunnelling optical near-field microscope (STOM) where the tip is used in the detection mode and the sample is illuminated by total internal reflection. The second configuration is an inverted STOM: the tip is used in the emission mode and the diffracted field is far-field detected in one direction. We present the models used to describe the two configurations and then explain the main lines of the formalism used to calculate the diffracted fields by a magneto-optical sample. PMID- 11388247 TI - Quantum theoretical approach to a near-field optical system. AB - The paper proposes a quantum theoretical formulation of an optical near-field system based on the projection-operator method. Special attention is paid to a nanometric probe tip-quantum mechanical sample system, whose interactions are essential for describing such phenomena as atom guidance and manipulation, or local excitation of a single quantum dot. The relationship to the virtual photon model--an intuitive model--is discussed, and the latter's empirical assumption of the Yukawa-type interaction between probe and sample is justified theoretically. Several applications of the theory are briefly outlined. PMID- 11388248 TI - Near-field optical excitation as a dipole-dipole energy transfer process. AB - The process of fluorescence excitation in the scanning near-field optical microscope (SNOM) is considered as a dipole-dipole resonance energy transfer process between a molecule under study and a SNOM aperture, which can be treated as a magnetic-type point dipole. It is shown that such an approach satisfactorily describes the conditions of the usual SNOM fluorescence experiments. Fluorescence excitation dependence on the polarization of the incident light and medium refraction index have been obtained. The equation to calculate the resonance dipole-dipole energy transfer radius (which is a natural unit of a SNOM's longitudinal resolution) is derived. Those cases where such a radius is of the order of the SNOM aperture, and thus single dipole, can strongly influence the radiation conditions are discussed briefly. PMID- 11388249 TI - Evanescent-wave scattering in near-field optical microscopy. AB - Extended Mie theory is used to investigate the scattering and extinction of evanescent waves by small spherical particles and aggregates of such particles. Metallic, dielectric and metal-coated dielectric particles are taken into consideration. In contrast to plane-wave excitation, p- and s-polarized spectra differ in the case of evanescent waves due to the inherent asymmetry of both polarizations. Furthermore, contributions from higher multipoles are strongly enhanced, compared with plane-wave excitation, and the enhancement factors are polarization dependent. The corresponding changes in the scattering and extinction spectra are most pronounced in cases where higher multipoles exhibit resonances in the spectral range considered. This applies, for example, to morphological resonances of dielectric particles with size parameters > 1. The effect of the surface, where the evanescent wave is generated by total internal reflection, on the scattering and extinction spectra is investigated via numerical field calculations employing the multiple multipole method. In an application to apertureless near-field optical microscopy, the variation of the scattered power is calculated when a silicon particle is scanned across a silver particle in the evanescent field. PMID- 11388250 TI - Spectroscopic study of the image formation in near-field microscopy, near an evanescent-homogeneous switching. AB - Near-field optical microscopes provide highly resolved images of various samples. However, images are difficult to interpret owing to their sensitivity to illumination conditions. Moreover, by contrast with classical microscopy, the near-field signal combines the contributions of evanescent and propagative modes. In this study, we present results of a spectroscopic study in near-field. Our purpose is to explain how a switching of one diffracted mode from homogeneous to evanescent can modify image formation. The main point is to establish a relation between the evanescence of one diffracted mode and the fringes that are often observed in near-field experimental images. Moreover, on a metallic sample, the possible occurrence of plasmon resonance contributes to image distortion in a mainly different way. We use a Fourier series Rayleigh 3D method to explain image formation. PMID- 11388251 TI - Polarization properties of the near-field intensity reflected by metallic and dielectric one-dimensional structures. AB - We investigate the state of polarization and near-field intensity distribution in the vicinity of rectangular groove objects ruled on metallic and dielectric materials. The sample is illuminated from the vacuum side by a linear combination of p- and s-polarised waves. Two rigorous methods of solution are used and compared in calculations of the total intensity at constant height when the light is incident normally onto the surface. Some calculations of the total intensity in the 'follow-the-profile mode' are also presented. It is shown that in the constant height mode, the contrast in the image can be reversed as the plane of observation moves away from the mean plane of the sample. We also found that the state of polarization depends strongly on the material and the distance to the plane of detection. PMID- 11388252 TI - Surface imaging in near-field optical microscopy by using the fluorescence decay rate: a theoretical study. AB - In this paper, we study the fluorescence decay rate of a molecule above a corrugated interface, and particularly the variations of the decay rate as a function of the lateral position of the molecule. As a first step, one has to determine the field diffracted by a corrugated interface when the incident field is the field emitted by a dipole. For this purpose, we have used a perturbative Rayleigh method, and we show that the decay rate variations can be connected to the surface profile via a transfer function. Some numerical calculations of this transfer function and of decay rate variation images are presented for dielectric and metallic samples. The visibility of the theoretical images is up to 20% and, moreover, resolution of the images is good enough to use the fluorescence lifetime of molecules as signal in a life-time scanning near-field optical microscope. The technical problems are discussed briefly. PMID- 11388253 TI - Fluorescence imaging with a laser trapping scanning near-field optical microscope. AB - We investigated fluorescence imaging using a near-field scanning optical microscope which uses a laser-stabilized gold nanoparticle as a near-field probe. This microscope is suitable for observations of biological specimens in aqueous solutions because the probe particle is held by a noncontact force exerted by a laser beam. Theoretical calculations based on Mie scattering theory are presented to evaluate the near-field enhancement by a gold particle of 40 nm diameter. We also present fluorescence images of a single fluorescent bead and discuss the near-field contribution to the fluorescence image in this type of microscope. PMID- 11388254 TI - Apertureless scanning near-field magneto-optical microscopy of magnetic multilayers. AB - We imaged magnetic domains in Pt/Co/Pt multilayers using an apertureless scanning near-field optical microscope operating in reflection mode. As the magneto optical effects are weak for this kind of structure, a polarization modulation technique with a photoelastic modulator was used to reveal the contrast between magnetic domains. In the case of a Pt/Co/Pt trilayer structure, a strong improvement in lateral resolution is observed compared with far-field magneto optical images and good sensitivity is achieved. In the case of a Pt/[Co/Pt]Pt multilayer structure, stripe domains of 200 nm width could be resolved, in good agreement with images obtained by magnetic force microscopy on the same structure. PMID- 11388255 TI - Characterization of reflection scanning near-field optical microscopy and scanning tunnelling optical microscopy/photon scanning tunnelling microscopy working in preliminary approach constant height scanning mode. AB - The resolution in near-field images is currently determined by the visual inspection of recorded images. One of the major questions in near-field optical microscopy is 'what resolution can be reached, the tip-to-sample distance being known?' This knowledge is critical when choosing the scanning step and the distance between the tip and the sample, in a preliminary scan. This preliminary scan is often the only way to detect the interesting parts of the sample, with limited risk of tip crash and topographical artefacts. The method proposed here needs two scans of the same area, of the same sample, in constant height mode, recorded at two tip-to-sample distances. The pseudotransfer function is the ratio of the Fourier transform of these two data maps. This function enables the evaluation of the limit of resolution. Theoretical considerations are introduced to assess the method. PMID- 11388256 TI - The tuning fork as sensor for dynamic force distance control in scanning near field optical microscopy. AB - The dynamic force distance control for scanning near-field optical microscopy on the basis of a tuning fork as piezoelectric force sensor is remarkably sensitive. In order to gain a better understanding of this sensitivity the vibrational properties of the tuning fork are modelled within the framework of two coupled harmonic oscillators. As a result, the effective force constant of the tuning fork at resonance frequency is determined. Furthermore, the influence of the additional mass by the attachment of the near-field probe is investigated. PMID- 11388257 TI - Optical characterization of probes for photon scanning tunnelling microscopy. AB - The photon scanning tunnelling microscope is a well-established member of the family of scanning near-field optical microscopes used for optical imaging at the subwavelength scale. The quality of the probes, typically pointed uncoated optical fibres, used is however, difficult to evaluate in a direct manner and has most often been inferred from the apparent quality of recorded optical images. Complicated near-field optical imaging characteristics, together with the possibility of topographically induced artefacts, however, has increased demands for a more reliable probe characterization technique. Here we present experimental results obtained for optical characterization of two different probes by imaging of a well-specified near-field intensity distribution at various spatial frequencies. In particular, we observe that a sharply pointed dielectric probe can be highly suitable for imaging when using p-polarized light for the illumination. We conclude that the proposed scheme can be used directly for probe characterization and, subsequently, for determination of an optical transfer function. which would allow one to deduce from an experimentally obtained image of a weakly scattering sample the field distribution existing near the sample surface in the absence of the probe. PMID- 11388258 TI - The height regulation of a near-field scanning optical microscope probe tip. AB - A nonoptical detection of the optical fibre tip has been developed. By detecting the output signal from a tiny piezoelectric detector attached to the vibrating fibre tip, the distance between the fibre tip and the sample has been successfully controlled. The frequency responses of the system composed of tip, the dither and the detector have been studied. The difference between the shear force detection and the tapping-mode detection is discussed. It is found that the shear force exerted on the tip reduces the vibration amplitude with an unvaried resonance frequency. However, in the tapping mode, the resonance frequency varies with the tip-sample distance as the force is exerted on the fibre tip only within a half period. This requires better adjustments for the tapping-mode detection. PMID- 11388259 TI - Optical-fibre scanning near-field optical microscope for cryogenic operation. AB - We have developed a new type of scanning near-field optical microscope (SNOM) utilizing optical fibres. The probe tip is controlled by shear force feedback with a fibre interferometer and signal light is collected directly by a multimode fibre. These features make the SNOM head more compact and less sensitive to vibration. Further advantages of this new type of SNOM are that it obviates the need for optical windows in the cryostat and offers easy optical alignment. PMID- 11388260 TI - High-speed scanning by dual feedback control in SNOM/AFM. AB - We have developed a high-speed scanning near-field optical microscope (SNOM)/atomic force microscope (AFM) system including dual feedback controllers. The system includes an additional piezoelectric actuator with fast response in the z direction and a correction circuit to eliminate unnecessary components from the feedback signal. From the measurement of a patterned chromium layer of 2 x 2 microm2 checks on a quartz glass plate, we confirmed that our system had more effective feedback control and faster scanning than current SNOM/AFM systems that use only a piezo-tube. The scanning speed of the present system was estimated to be about five times faster than that of current SNOM/AFM systems. PMID- 11388261 TI - Femtosecond near-field scanning optical microscopy. AB - We have developed an instrument for optically measuring carrier dynamics in thin film materials with approximately 150 nm lateral resolution, approximately 250 fs temporal resolution and high sensitivity. This is accomplished by combining an ultrafast pump-probe laser spectroscopic technique with a near-field scanning optical microscope. A diffraction-limited pump and near-field probe configuration is used, with a novel detection system that allows for either two-colour or degenerate pump and probe photon energies, permitting greater measurement flexibility than that reported in earlier published work. The capabilities of this instrument are proven through near-field degenerate pump-probe studies of carrier dynamics in GaAs/AIGaAs single quantum well samples locally patterned by focused ion beam (FIB) implantation. We find that lateral carrier diffusion across the nanometre-scale FIB pattern plays a significant role in the decay of the excited carriers within approximately 1 microm of the implanted stripes, an effect which could not have been resolved with a far-field system. PMID- 11388262 TI - Nano-slit probes for near-field optical microscopy fabricated by focused ion beams. AB - The near-field probes described in this paper are based on metallized non-contact atomic force microscope cantilevers made of silicon. For application in high resolution near-field optical/infrared microscopy, we use aperture probes with the aperture being fabricated by focused ion beams. This technique allows us to create apertures of sub-wavelength dimensions with different geometries. In this paper we present the use of slit-shaped apertures which show a polarization dependent transmission efficiency and a lateral resolution of < 100 nm at a wavelength of 1064 nm. As a test sample to characterize the near-field probes we investigated gold/palladium structures, deposited on an ultrathin chromium sublayer on a silicon wafer, in constant-height mode. PMID- 11388263 TI - A novel fabrication method for fluorescence-based apertureless scanning near field optical microscope probes. AB - We report a novel method for the fabrication of probes with localized sub wavelength fluorescing media at their extremities. We present our first results and discuss future plans to extend this technique to the systematic fabrication of fluorescent probes for apertureless scanning near-field optical microscopy. PMID- 11388264 TI - Modified fabrication process for aperture probe cantilevers. AB - We report the development of cantilever- and fibre-based probes for scanning near field optical microscopy. Both probe concepts rely on the integration of a microfabricated aperture tip with reproducible optical and mechanical properties. Numerical calculations were carried out using a finite integration code to investigate the polarization-sensitive transmission behaviour of aperture tips. In order to establish technological guidelines for the optimization of the properties of the optical tip the distinct influence of the tip geometry on the intensity distribution in the vicinity of the aperture is studied in detail. PMID- 11388265 TI - Coaxial probes for scanning near-field microscopy. AB - This paper deals with the development of coaxial aperture tips integrated in a cantilever probe for combined scanning near-field infrared microscopy and scanning force microscopy. A fabrication process is introduced that allows the batch fabrication of hollow metal aperture tips integrated on a silicon cantilever. To achieve the coaxial tip arrangement a metal rod is deposited inside the hollow tip using the focused ion beam technique. Theoretical calculations with a finite integration code were performed to study the transmission characteristics of coaxial tips in comparison with conventional aperture probes. In addition, the influence of the geometrical design parameters of the coaxial probe on its optical behaviour is investigated. PMID- 11388267 TI - Detection of an infrared near-field optical signal by attaching an infrared excitable phosphor to the end of a photocantilever. AB - To improve the signal-to-noise ratio of near-field scanning optical microscopy, we propose attaching an infrared-excitable phosphor (IEP) to a photocantilever. One source of noise is the light scattered from locations on the sample surface other than that of the probe tip. By detecting only the light scattered from the tip, we can obtain a near-field optical signal without noise. We attached an IEP particle to a photocantilever to convert infrared light to visible light and we used 1550-nm infrared illumination, so the light scattered from the sample was only infrared. The silicon photodiode of the photocantilever is 10(6) times less sensitive to infrared light than to visible light. As a result, only the converted visible light from the IEP particle, i.e. the signal containing the near-field optical information from the tip, was detected. We verified that the photocantilever detected the signal in the evanescent light produced by infrared illumination and that the detected signal was the light converted by the IEP. The experimental results show the feasibility of detecting infrared light and not the background light through the use of the IEP. PMID- 11388266 TI - Diffraction of circularly polarized light from near-field optical probes. AB - Diffracted fields from 100-nm aperture near-field scanning optical microscopy (NSOM) probes and uncoated tapered fibres are measured and analysed. Using a solid angle scanner, the two-dimensional intensity distribution and polarization state of the diffracted light are resolved experimentally. Polarization analyses show that circularly polarized input light does not maintain its polarization state for all diffraction angles, and is completely filtered into linearly polarized light at large polar diffraction angles. This drastic decomposition originates from the vector nature of light diffracted by the sub-wavelength aperture. There is a fundamental difficulty in generating circularly polarized light near the aperture of NSOM probes owing to polarization-dependent diffraction in the near-field regime. This is illustrated by the Bethe-Bouwkamp model using circularly polarized input light. PMID- 11388268 TI - Towards better scanning near-field optical microscopy probes--progress and new developments. AB - Several approaches are described with the aim of producing near-field optical probes with improved properties. Focused ion beam milling allows the fabrication of small apertures in a controlled fashion, resulting in probes with excellent polarization properties and increased transmission. Microfabrication processes are described that allow the production of apertures of 30-50 nm, facilitating the mass-fabrication of apertured tip structures that can be used in a combined force/near-field optical microscope. Finally, possible future developments are outlined. PMID- 11388269 TI - Evaluation of nano-optical probe from scanning near-field optical microscope images. AB - We studied a nanometre-sized optical probe in a scanning near-field optical microscope. The probe profile is determined by using a knife-edge method and a modulated transfer function evaluation method which uses nanometre-sized line-and space tungsten patterns (with spaces 1 microm to 50 nm apart) on SiO2 substrates. The aluminium-covered, pipette-pulled fibre probe used here has two optical probes: one with a large diameter (350 nm) and the other with a small diameter (10 nm). The small-diameter probe has an optical intensity approximately 63 times larger than that of the large-diameter probe, but the power is about 1/25 of that of the large probe. PMID- 11388270 TI - Polarization properties of bent-type optical fibre probe for magneto-optical imaging. AB - Quantitative evaluation of magneto-optical parameters is necessary in order to apply scanning near-field optical microscope (SNOM) technology to the study of magnetism on the mesoscopic scale. For this purpose, quantitative knowledge of polarization transmission properties through an optical fibre probe is required. We therefore determined the Stokes parameters of the bent-type optical fibre probe that is used as a cantilever for atomic force microscope operation in our SNOM system. As a result, it is found that the degree of polarization is maintained in the light emitted from the probe, although the probe acts as if it were a wave plate. This anisotropic polarization state of the light emitted from the probe was compensated for by using a Berek compensator placed in front of the fibre coupler. PMID- 11388271 TI - Brighter near-field optical probes by means of improving the optical destruction threshold. AB - The optical destruction thresholds of conventionally etched and tube-etched near field optical probes were measured. One of the main advantages of tube-etched tips is their smooth glass surface after taper formation. Presumably for this reason, a destruction limit of over 120 microJ was obtained, almost twice as large as that of the rougher, conventionally etched fibre probes. The use of additional adhesion layers (Ti. Cr, Co and Ni) between the glass surface and the aluminium coating produced, especially for tube-etched tips, a significant increase in the optical destruction threshold. With increasingly thin metal coatings, the use of a protection coating that prevents corrosion during aging is recommended. An additional increase in optical stability was achieved by applying mixed-metal coatings: alternating thin titanium and thick aluminium layers yielded fibre probes with superior properties that achieved average optical destruction thresholds of > 270 microJ. This is an increase in stability of > 400% compared with conventionally fabricated near-field optical tips. PMID- 11388272 TI - Dynamic etching method for fabricating a variety of tip shapes in the optical fibre probe of a scanning near-field optical microscope. AB - A novel etching method for an optical fibre probe of a scanning near-field optical microscope (SNOM) was developed to fabricate a variety of tip shapes through dynamic movement during etching. By moving the fibre in two-phase fluids of HF solution and organic solvent, the taper length and angle can be varied according to the movement of the position of the meniscus on the optical fibre. This method produces both long (sharp angle) and short (wide angle) tapered tips compared to tips made with stationary etching processes. A bent-type probe for a SNOM/AFM was fabricated by applying this technique and its throughput efficiency was examined. A wide-angle probe with a 50 degrees angle at the tip showed a throughput efficiency of 3.3 x 10(-4) at a resolution of 100 nm. PMID- 11388273 TI - Optical microcantilever consisting of channel waveguide for scanning near-field optical microscopy controlled by atomic force. AB - We develop a novel optical microcantilever for scanning near-field optical microscopy controlled by atomic force mode (SNOM/AFM). The optical microcantilever has the bent channel waveguide, the corner of which acts as aperture with a large tip angle. The resonance frequency of the optical microcantilever is 9 kHz, and the spring constant is estimated to be 0.59 N/m. The optical microcantilever can be operated in contact mode of SNOM/AFM and we obtain the optical resolution of about 200 nm, which is as same size as the diameter of aperture. We confirm that the throughput of optical microcantilever with an aperture of 170 nm diameter would be improved to be more than 10(-5). PMID- 11388274 TI - Time-resolved near-field optics: exciton transport in semiconductor nanostructures. AB - We present a systematic, temperature-dependent study of excitonic real-space transfer into single GaAs quantum wires using time-resolved low-temperature near field luminescence spectroscopy. Excitons generated by local short pulse optical excitation in a 250 nm spot undergo diffusive transport over a length of several micrometres and are subsequently trapped into the quantum wire by optical phonon emission. The effect of local energy barriers in the vicinity of the quantum wire on the real-space transfer dynamics is monitored directly by mapping the time resolved quantum wire luminescence. Experiments at variable temperatures are compared to numerical simulations based on drift-diffusive model calculations, and the spatio-temporal evolution of the two-dimensional exciton distribution within the nanostructure is visualized. PMID- 11388275 TI - Photocurrent near-field microscopy of Schottky barriers. AB - We used a combination of internal photoemission and of near-field optical microscopy (SNOM) to study the lateral variations in solid interface properties such as energy barriers and electron-hole recombination. In particular we investigated the fully formed Pt-GaP, Au-GaAs, Au-SiNx-GaAs and PtSi-Si Schottky barriers. Our approach enabled us to measure large lateral variations in the photocurrent with spatial resolution on the nanometric scale. Due to the ability of SNOM to supply parallel topographic information, we observed photocurrent variations from zone to zone that only correlated in a few cases with local variations in surface morphology. We assigned the uncorrelated fluctuations to local variations in the interface stoichiometry, the presence of interface states induced by the metallic overlayer and to defect states at the junction. Furthermore, by tuning the photon energy and applied bias we were able to measure the surface distribution of the diffusion length. PMID- 11388277 TI - Spectrally resolved cathodoluminescence analyses in the optical near-field. AB - By implementing a scanning near-field optical microscope into the specimen chamber of a scanning electron microscope, cathodoluminescence can be locally detected in the optical near-field. The achievable spatial resolution in this set up is only limited by the size of the aperture in a coated fibre probe and its separation from the sample, rather than by the energy dissipation volume of the primary electrons and diffusion processes of excess carriers inside the specimen. We demonstrate how electronically active defects in polycrystalline diamond can be distinguished and localized with sub-wavelength lateral resolution by spectral filtering of the cathodoluminescence signal. PMID- 11388276 TI - Inducing superconductivity at a nanoscale: photodoping with a near-field scanning optical microscope. AB - The local modification of an insulating GdBa2Cu3O6.5 thin film, made superconducting by illumination with a near-field scanning optical microscope (NSOM), is reported. A 100-nm aperture NSOM probe acts as a sub-wavelength light source of wavelength lambda(exc) = 480-650 nm, locally generating photocarriers in an otherwise insulating GdBa2-Cu3O6.5 thin film. Of the photogenerated electron-hole pairs, electrons are trapped in the crystallographic lattice, defining an electrostatic confining potential to enable the holes to move. Reflectance measurements at lambda = 1.55 microm at room temperature show that photocarriers can be induced and constrained to move on a approximately 200 nm scale for all investigated lambda(exc). Photogenerated wires present a superconducting critical temperature Tc= 12 K with a critical current density Jc = 10(4) A cm(-2). Exploiting the flexibility provided by photodoping through a NSOM probe, a junction was written by photodoping a wire with a narrow (approximately 50 nm) under-illuminated gap. The strong magnetic field modulation of the critical current provides a clear signature of the existence of a Josephson effect in the junction. PMID- 11388278 TI - Photoluminescence imaging of phosphor particles using near-field optical microscope with UV light excitation. AB - We have developed fibre probes suitable for 325 nm UV light excitation and a photoluminescence near-field scanning optical microscope (NSOM) and demonstrated the photoluminescence imaging of phosphor BaMgAl10O17:Eu2+ (BAM) particles. The probe was fabricated by a two-step-etching method that we developed. The probe had a large taper angle at the top of the probe and a small taper angle at the root. The NSOM image was different from the topographical structure but roughly reflected the corresponding features of the particles. The inhomogeneity of the photoluminescence intensity between BAM particles was observed in the NSOM image. The photoluminescence intensity with various bandpass filters showed differences between the individual particles, which means that they have different spectra. PMID- 11388279 TI - Polarization effects in near-field excitation-collection probe optical microscopy of a single quantum dot. AB - We solve numerically the three-dimensional vector form of Maxwell's equation for the situation of near-field excitation and collection of luminescence from a single quantum dot, using a scanning near-field optical fibre probe with subwavelength resolution. We highlight the importance of polarization-dependent effects in both the near-field excitation and collection processes. Applying a finite-difference time domain method, we calculate the complete vector fields emerging from a realistic probe structure which is in close proximity to a semiconductor surface. We model the photoluminescence from the quantum dot in terms of electric dipoles of different polarization directions, and determine the near-field luminescence images of the dot captured by the same probe. We show that a collimating effect in the high index semiconductor significantly improves the spatial resolution in the excitation-collection mode. We find that the spatial resolution, image shape and collection efficiency of near-field luminescence imaging strongly depend on the polarization direction as represented by the orientation of the radiating electric dipoles inside the quantum dot. PMID- 11388280 TI - Near-field second harmonic imaging of lead zirconate titanate piezoceramic. AB - Near-field optical microscopy has been used to image variations in local optical second harmonic generation (SHG) from the surface of lead zirconate titanate (PZT) piezoceramic. As PZT ceramic is a strongly scattering medium. SHG occurs in the thin layer near the surface of the ceramic. Thus, individual crystalline grains and grain boundaries located near the surface are the main features visible in the images. In general, this technique allows us to determine the local poling direction of individual submicrometre-sized crystalline grains of ceramic by near-field SH imaging at different angles of incidence and polarization states of the fundamental excitation light. In some cases 'hot spots' of submicrometre size showing enhanced SHG have been observed. This enhancement is believed to result from local cavity resonances. PMID- 11388281 TI - Thickness measurement of thin dielectric films by evanescent total reflection fluorescence. AB - The electric field of an evanescent wave generates fluorescence in the interface between a dielectric surface and an adjacent, fluorescing, medium of lower refractive index. The difference between the fluorescing signals from covered and noncovered surfaces enables nondestructive measurement of the film thickness to be made in the range 1-15 nm. PMID- 11388282 TI - Near-field mapping of the emission distribution in semiconductor microdiscs. AB - We have used a scanning near-field optical microscope to study the fluorescent light distribution in the near- and far-fields with two types of microdiscs, InGaP and GaN, fabricated in our laboratory. The InGaP microdisc has a radius of 2.5-5.0 microm, a thickness of 0.15-0.2 microm and a circular shape and the GaN disc has a radius of 5-8 microm with a thickness of 0.5-2 microm. Spontaneous emission enhancement in these microdiscs has been observed with emitting wavelengths of 650 and 550 nm respectively In both types of microdisc, the whispering-gallery mode (WCM) has been observed on the top surface using near field optical and far-field microscopic methods. However, due to the different disc structures and optical confinements, the light distributions of the type types of disc are quite different. In the case of the InGaP disc, WGM is the dominant mode with a mixture of other modes. Interference-like ring intensities have been observed both inside the disc surface and outside, with a period ratio of 1:2. In addition, the propagating waves emitted from the side of the disc have been collected for the first time by using near-field optical microscopy. A theoretical calculation based on the theory of optical modes in microdisc lasers confirmed this observation. It also predicted the behaviour of the electric field distribution (transverse electric) inside and outside the disc, as well as the period of the wave propagation. In contrast, the near-field mapping of the GaN fluorescence showed not only a ring-like emission intensity along the circumference of the disc, an indication of WGM, but also an even intensity distribution inside the disc. This can be explained as the combination of the WGM with the Fabry-Perot mode of multi-reflection between the GaN layer and the substrate. The results also demonstrate the potential application of near-field optics to explore the light emission mode of a microdisc on a nanometre scale. PMID- 11388283 TI - Interactions between hydrophobic and hydrophilic silicon surfaces using a tapered probe in near-field scanning optical microscopy. AB - We examined the tip-sample interaction of a Si(111) surface processed with HF solution. The interaction length of the probe with its hydrophobic surface is approximately 30, and the frequency responses show an interaction force of elastic type. By contrast, the damping curve on a hydrophilic surface, Si with thin oxide, operates on a longer length of 100. Here, the frequency responses indicate an interaction force of the viscous type. PMID- 11388285 TI - Domain formation in thin lipid films probed with near-field scanning optical microscopy. AB - High-resolution near-field scanning optical microscopy (NSOM) fluorescence and topographic images of L-alpha-dipalmitoylphosphatidylcholine (DPPC) monolayers doped with a fluorescent dye are presented. DPPC monolayers are deposited onto mica substrates from the air-water interface at several surface pressures using the Langmuir-Blodgett technique. Sub-diffraction limit phase domain structures are observed in both fluorescence and topographic NSOM images of the lipid films. The morphology of the resulting monolayers depends strongly on the surface pressure and composition of the subphase used in the film transfer. Mechanisms for lipid domain formation and growth are discussed. PMID- 11388286 TI - Near-field scanning optical microscopy studies of L-alpha dipalmitoylphosphatidylcholine monolayers at the air-liquid interface. AB - The phase structure in L-alpha-dipalmitoylphosphatidylcholine-20 mol% fluorescent 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate Langmuir monolayers dispersed on a 2 M sucrose solution subphase is studied with near field scanning optical microscopy (NSOM). Cantilevered NSOM probes operating in a tapping-mode feedback or an optical interferometric feedback mode are capable of tracking the air-sucrose solution interface. At the micrometre scale, the NSOM fluorescence images reveal lipid domain features similar to those observed previously in supported Langmuir-Blodgett (LB) monolayers. At the submicrometre scale, the small nanometric lipid islands seen in LB films are not observed at the air-sucrose interface. This supports a mechanism in which domain formation in LB films can be induced by means of the transfer process onto the solid support. Progress towards extending these studies to films at the air-water interface using the optical interferometric feedback method is also discussed. PMID- 11388288 TI - Evanescent field excitation and measurement of dye fluorescence in a metallic probe near-field scanning optical microscope. AB - We introduce a method of dye fluorescence excitation and measurement that utilizes a near-field scanning optical microscope (NSOM). This NSOM uses an apertureless metallic probe, and an optical system that contains a high numerical aperture (NA) objective lens (NA= 1.4). When the area which satisfies NA < 1 is masked, the objective lens allows for the rejection of possible transmitted light (NA < 1) through the sample. In such conditions, the focused spot consists of only the evanescent field. We found that this NSOM system strongly reduces the background of the dye fluorescence and allows for the measurement of the fluorescence intensity below the diffraction limit of the excitation source. PMID- 11388287 TI - Imaging of various surface properties of fluorescently labelled phospholipid Langmuir-Blodgett films with a combined scanning probe microscope. AB - We present results of phase separation of a single-component system of 1,2 dihexadecanoyl-sn-glycero-3-phospho-[N-(4-nitrobenz)-2-oxa-1,3 diazolyl]ethanolamine in which a liquid-condensed (LC) phase co-exists with a liquid-expanded (LE) phase. Domain formation in the co-existence region was studied using a newly developed combined scanning near-field optical microscope atomic force microscope (SNOM-AFM). We demonstrate for the first time that the topographic, friction, fluorescence and surface potential distributions for a phase-separated single-component Langmuir-Blodgett film between the LE and LC phases can be simultaneously observed using the SNOM-AFM with a thin-step etched optical fibre probe. PMID- 11388289 TI - Single molecule mapping of the optical field distribution of probes for near field microscopy. AB - The most difficult task in near-field scanning optical microscopy (NSOM) is to make a high quality subwavelength aperture probe. Recently, we have developed high definition NSOM probes by focused ion beam (FIB) milling. These probes have a higher brightness, better polarization characteristics, better aperture definition and a flatter end face than conventional NSOM probes. We have determined the quality of these probes in four independent ways: by FIB imaging and by shear-force microscopy (both providing geometrical information), by far field optical measurements (yielding throughput and polarization characteristics), and ultimately by single molecule imaging in the near-field. In this paper, we report on a new method using shear-force microscopy to study the size of the aperture and the end face of the probe (with a roughness smaller than 1.5 nm). More importantly, we demonstrate the use of single molecules to measure the full three-dimensional optical near-field distribution of the probe with molecular spatial resolution. The single molecule images exhibit various intensity patterns, varying from circular and elliptical to double arc and ring structures, which depend on the orientation of the molecules with respect to the probe. The optical resolution in the measurements is not determined by the size of the aperture, but by the high optical field gradients at the rims of the aperture. With a 70 nm aperture probe, we obtain fluorescence field patterns with 45 nm FWHM. Clearly, this unprecedented near-field optical resolution constitutes an order of magnitude improvement over far-field methods like confocal microscopy. PMID- 11388290 TI - Laser scanning gradient index optics microscope: submicrometre scale spectroscopy and imaging at cryogenic temperatures. AB - A laser scanning far-field optical microscope for low-temperature imaging and spectroscopy based on gradient index optics is presented. A rod-shaped gradient index microlens is used as a zero-working-distance solid immersion objective lens. The obtained lateral resolution is 310 nm of the FWHM at a wavelength of 545 nm. A laser scanning mechanism located outside an optical cryostat enables one to achieve large scanning ranges independent of temperature. The use of the microscope for submicrometre-scale spectroscopy and low-temperature photochemistry performed on molecular J aggregates in thin polymer films is presented. PMID- 11388291 TI - Field enhancement and apertureless near-field optical spectroscopy of single molecules. AB - We report on fluorescence enhancement in near field optical spectroscopy by apertureless microscopy. Our apertureless microscope is designed around a confocal fluorescence microscope associated with an AFM head. First, we show that the confocal microscope alone allows single molecule imaging and single molecule fluorescence analysis. When associated with the AFM head, we demonstrate, for the first time to our knowledge, that single molecule fluorescence is enhanced under the silicon tip. We tentatively attribute this effect to field enhancement under the tip. PMID- 11388292 TI - The depolarization near-field scanning optical microscope: comparison of experiment and theory. AB - The depolarization near-field scanning optical microscope allows for sub wavelength optical resolution with uncoated optical fibre tips. We show by a simple thought experiment, by numerical solution of the Maxwell equations for a realistic geometry, and by direct experiments, that this novel apparatus images gradients of the dielectric function of a sample in the direction of the incident electric field vector. The near-field images obtained from experiment and theory agree well. PMID- 11388293 TI - Magneto-optical contrast in near-field optics. AB - We propose a simple calculation of near-field magneto-optical (MO) images based on the beam propagation method. We calculate both Faraday rotation and circular dichroism contrasts of planar magnetic structures such as as-grown thin films and ion-irradiated samples. High-contrast near-field MO images are obtained, in good agreement with our experimental observations. PMID- 11388294 TI - Polarization contrast in reflection near-field optical microscopy with uncoated fibre tips. AB - Using cross-hatched, patterned semiconductor surfaces and round 20-nm-thick gold pads on semiconductor wafers, we investigate the imaging characteristics of a reflection near-field optical microscope with an uncoated fibre tip for different polarization configurations and light wavelengths. It is shown that cross polarized detection allows one to effectively suppress far-field components in the detected signal and to realize imaging of optical contrast on the subwavelength scale. The sensitivity window of our microscope, i.e. the scale on which near-field optical images represent mainly optical contrast, is found to be approximately 100 nm for light wavelengths in the visible region. We demonstrate imaging of near-field components of a dipole field and purely dielectric contrast (related to well-width fluctuations in a semiconductor quantum well) with a spatial resolution of approximately 100 nm. The results obtained show that such a near-field technique can be used for polarization-sensitive imaging with reasonably high spatial resolution and suggest a number of applications for this technique. PMID- 11388296 TI - Mid-infrared scanning near-field optical microscope resolves 30 nm. AB - We explore the performance of a scanning near-field infrared microscope, which works by scattering tightly focused CO2 laser radiation (lambda = 10 microm) from the apex of a metallized atomic force microscope tip. The infrared images of test samples prove a spatial resolution of 30 nm and are free of topographical and inertial artefacts, thus they should be of great interest for practical applications. We also observe that the infrared contrast vanishes when the input beam polarization is orthogonal to the tip axis, in accordance with theoretical expectations for a mechanism of longitudinal field interaction. PMID- 11388295 TI - Magnetic domain imaging with a scanning near-field optical microscope using a modified Sagnac interferometer. AB - We report on the combination of a scanning near-field optical microscope and a modified Sagnac interferometer for magnetic-domain imaging in the reflection mode. The Sagnac interferometer is used for detection of the magneto-optical Kerr effect. Since the interferometer is inherently insensitive to polarization changes caused by topography effects, magnetic-domain imaging is not limited to samples with flat surfaces. In this way, it is possible to image magnetic bits written on the tracks of a magneto-optical disc that has a rather pronounced surface profile. PMID- 11388297 TI - Interferometric measurement of femtosecond optical pulses emitted from a fibre probe. AB - An interferometric technique is applied to a temporal analysis of the far-field radiation from a near-field optical fibre probe. A balanced homodyne scheme combined with lock-in detection at dual harmonics allows one to evaluate the temporal form of weak signal pulses, even in the presence of some environmental vibrations that disturb the optical interference. The transmission properties of a small aperture are formulated theoretically. It is shown that the sub wavelength aperture should not distort the femtosecond optical pulses temporally, which is consistent with the experimental observation. PMID- 11388298 TI - Spectroscopy with scanning near-field optical microscopy using photon tunnelling mode. AB - We have demonstrated Raman spectroscopy using scanning near-field optical microscopy (SNOM). Photon tunnelling mode was employed, in which the sample is illuminated using an attenuated total reflection (ATR) configuration and the evanescent wave perturbed by the sample is picked up by a sharpened optical fibre probe. By this experimental arrangement Raman scattering from the optical fibre probe was greatly reduced, therefore we were able to excite the sample using more intense laser light compared to the illumination mode SNOM. Raman spectra of copper phthalocyanine (CuPc) were obtained in the off-resonance condition and without using surface-enhanced Raman scattering (SERS). PMID- 11388299 TI - Ultrafast nonlinear sub-wavelength solid immersion spectroscopy at T = 8 K: an alternative to nonlinear scanning near-field optical microscopy. AB - Pump-probe measurements were performed with a subwavelength spatial resolution of 355 nm and a temporal resolution of 130 fs in a multiple quantum well sample at T= 8 K. A solid immersion lens was used to increase the spatial resolution to 0.41 lambda, demonstrating that the limit of conventional microscopy was surpassed. PMID- 11388301 TI - Near-field imaging of surface-enhanced second harmonic generation. AB - Surface-enhanced second harmonic generation from individual topographical defects of an otherwise flat gold film and from metal-coated diffraction gratings was measured using a near-field optical microscope. Experimentally measured second harmonic field distributions were compared with theoretical calculations. PMID- 11388302 TI - Near-field optical photomask repair with a femtosecond laser. AB - We present a high-resolution near-field optical tool designed for repair of opaque defects in binary photomasks. Both instrument design and near-field imaging and patterning results will be presented. Designed for ablative processing of thin metal films, the MR-100 incorporates an industrial amplified femtosecond laser, third harmonic generator and built-in autocorrelator. The ultrashort duration of the femtosecond pulses enables the tool to remove chrome layers with negligible damage to the surrounding metal or the underlying quartz substrate. The micropipette based near-field writing head can deliver power densities of hundreds of GW/cm2 to spots of several hundred nanometres and below. Repairs on sample masks will be presented and the repair quality will be discussed. PMID- 11388303 TI - Reflection mode scanning near-field optical microscopy analyses of integrated devices. AB - Ultra-large-scale integrated devices have been investigated by a reflection mode scanning near-field optical microscope designed for semiconductor analyses. Although it could be found that imaging the reflectivity of metal structures buried underneath thin, optically transparent passivation layers is practicable, shading of the reflected light by the SNOM probe complicated the interpretability of the achieved results. This issue has been overcome by using the SNOM probe as both illumination source and detector, simultaneously. The application of focused ion beam milling to the probes has allowed the increase of the transmittivity of the probes to such an extent that coated fibres could be utilized and the interpretability of the results could be enhanced. Furthermore, the investigated structures are of great interest for an investigation of z-motion artefacts, as the presence of the polished passivation layers allows topographical influences to be distinguished from pure optical contrast. PMID- 11388304 TI - Nanometric patterning of zinc by optical near-field photochemical vapour deposition. AB - A new technique, optical near-field photochemical vapour deposition (NFO-PCVD) enables maskless production of nanometric structures with controllable size, chemical composition and morphology. By placing a near-field optical microscope inside the reaction chamber for photochemical vapour deposition we have deposited nanoscale metal patterns. We demonstrate for the first time, successfully deposited in the near-field region, lines of metallic zinc with the observed stripe width of 20 nm. PMID- 11388305 TI - Imaging of optical disc using reflection-mode scattering-type scanning near-field optical microscopy. AB - A phase-change optical disc was observed using a reflection-mode scattering-type scanning near-field optical microscope (RS-SNOM). In an a.c.-mode SNOM image, the 1.2 microm x 0.6 microm recording marks were successfully observed although the data were recorded on the groove. In contrast, no recording marks could be resolved in a d.c.-mode SNOM image. These results are in good agreement with those from a numerical simulation using the finite difference time domain method. The resolution was better than 100 nm with a.c.-mode SNOM operation and the results indicate that recording marks in phase-change optical media can be directly observed with the RS-SNOM. PMID- 11388307 TI - Near-field optical microscopy of localized excitations on rough surfaces: influence of a probe. AB - Starting from the general principles of near-field optical microscopy. I consider the influence of a probe when being used to image localized dipolar excitations and suggest a way of evaluating the perturbation thus introduced. Using the rigorous microscopic (electric) point-dipole description, I calculate the self consistent field intensity at the site of a probe dipole scanning over resonantly interacting object dipoles and show that the intensity distribution deviates from that existing in the absence of a probe. I demonstrate that this difference increases with an increase in the polarizability of the probe dipole. resulting eventually in a completely different intensity distribution. The calculations also show that the perturbation of the intensity distribution due to the presence of a probe decreases with an increase in the probe-sample distance. In order to evaluate the degree of perturbation. I suggest comparing the images obtained at different probe-sample distances. Finally, I formulate a simple rule of thumb that allows one to roughly estimate the probe-sample coupling when imaging localized excitations. PMID- 11388306 TI - Fabrication and observation of a standard sample for near-field optical microscopy. AB - We fabricated a standard sample for a near-field optical microscope using scanning probe lithography. The sample contains a wedged pattern, which allows the measurement of various sizes within one image. The optical resolution of our near-field optical microscope has been evaluated as 40 nm, which was obtained by measuring the narrowest separable gap width of the wedged pattern. Thus a standard sample containing the wedged pattern enables clear evaluation of the resolution. PMID- 11388308 TI - Surface-polariton propagation for scanning near-field optical microscopy application. AB - Surface plasmon-, phonon- and exciton-polaritons exist on specific materials in specific spectral regions. We assess the properties of such travelling surface bound electromagnetic waves relevant for scanning near-field optical microscopy applications, i.e. the tightness of surface binding, the attenuation, the phase velocity and the coupling with free-space electromagnetic waves. These quantities can be directly determined by photographic imaging of surface plasmon- and surface phonon-polaritons, in both the visible and mid-infared regions. Focusing of mid-infrared surface plasmons is demonstrated. Surface waveguides to transport and focus photons to the tip of a scanning near-field probe are outlined. PMID- 11388309 TI - Plasmon transmissivity and reflectivity of narrow grooves in a silver film. AB - Surface plasmon (SP) reflectivity and transmissivity of narrow grooves in silver films are studied. The SP source is the probe of a scanning near-field optical microscope. Locally detected leakage radiation from the SP provides detailed information on the paths of SP propagation, in particular the influence of perturbations. Global detection provides representative average data on the SP properties of a given metal film and its structures. A groove of 200 nm width, for instance, reflects/transmits about 15%/80% of 'blue-green' SP radiation at normal incidence. PMID- 11388310 TI - Near-field optical studies of local photomodification in nanostructured materials. AB - Fractal aggregates of silver nanoparticles are studied experimentally using atomic force microscopy and photon scanning tunnelling microscopy. Large changes in the near-field optical response of fractal aggregates are observed after the irradiation of samples with nanosecond laser pulses. The threshold energy density for photomodification using a 532 nm laser is measured to be 9 mJ cm(-2). It is shown that photomodification-induced changes in the local optical response can be two orders of magnitude larger than changes in far-field absorption. PMID- 11388311 TI - The interaction of surface plasmon polaritons with a silver film edge. AB - A prism coupling arrangement is used to excite surface plasmons at the surface of a thin silver film and a photon scanning tunnelling microscope is used to detect the evanescent field above the silver surface. Excitation of the silver/air mode of interest is performed at lambda1 = 632.8 nm using a tightly focused beam, while the control of the tip is effected by exciting a counter-propagating surface plasmon field at a different wavelength, lambda2 = 543.5 nm, using an unfocused beam covering a macroscopic area. Propagation of the red surface plasmon is evidenced by an exponential tail extending away from the launch site, but this feature is abruptly truncated if the surface plasmon encounters the edge of the silver film--there is no specularly reflected 'beam'. Importantly, the radiative decay of the surface mode at the film edge is observable only at larger tip-sample separations, emphasizing the importance of accessing the mesoscopic regime. PMID- 11388312 TI - Interview between Lindsay Peer and Bob Turney. PMID- 11388313 TI - A 67-year-old woman with Parkinsonism. PMID- 11388314 TI - [Oral health of community-dwelling elderly. 2. The subjective aspect of oral health]. AB - In this study oral health of elderly persons as perceived by the subjects was evaluated. Phenomena often mentioned were problems with chewing and biting, dry mouth, retention of food particles between teeth or below the prosthesis, (in edentates) sensitivity of teeth for warm or cold foods and (in edentates) lack of retention of the lower prosthesis. In a lot of persons these problems affect quality of life. PMID- 11388315 TI - [Three years of NTvT(Dutch Journal of Dentistry) online]. PMID- 11388316 TI - Secondary maternal cytomegalovirus infection--A significant cause of congenital disease. PMID- 11388317 TI - Cholesterol guidelines debate. PMID- 11388318 TI - Management of minor closed head injury in children. PMID- 11388319 TI - Computed tomography in diagnosing suspected appendicitis. PMID- 11388320 TI - A long letter and an even longer reply about autism magnetoencephalography and electroencephalography. PMID- 11388321 TI - Diagnostic value of cytokine measurement in cerebrospinal fluid in children with central nervous system tuberculosis. PMID- 11388322 TI - Echocardiography in healthy children. PMID- 11388323 TI - Analyzing the cost of neonatal screening for congenital adrenal hyperplasia. PMID- 11388324 TI - Diagnosis and evaluation of the child with attention-deficit/hyperactivity disorder. PMID- 11388325 TI - Can mixed strategies be stable in asymmetric games? AB - Selten (1980, J. theor. Biol. 84, 93(N)/01) has shown that mixed strategies cannot be evolutionarily stable in asymmetric games. Because every interaction features some asymmetry, this result apparently precludes mixed strategies in an evolutionary setting. In Maynard Smith's Hawk-Dove game (1982, Evolution and the theory of games (UP-Cambridge), for example, Selten's result restricts attention to pure-strategy evolutionarily stable outcomes in which the animals use the ability to condition their actions on asymmetries to coordinate, with one playing Hawk and one playing Dove, and with conflicts in which both animals play Hawk never arising. This result contrasts with the intuition that the mixed equilibrium of the Hawk-Dove game captures important aspects of many animal interactions, including the possibility of conflict. In this paper, we follow Eshel and Sansone (1995, J. theor. Biol. 177, 341-356) in enriching Selten's model to incorporate an important aspect of animal interactions, namely that payoffs and asymmetries may both be imperfectly observed. In the richer model, we find conditions under which effectively mixed strategies are stable in asymmetric games, as well as conditions under which they are not stable. Behavior will be conditioned on asymmetries, leading to pure-strategy equilibria in which conflict is avoided, when there are relatively large, observable asymmetries and small observable variations in payoffs. Under opposite conditions, evolutionarily stable equilibria will appear that are effectively mixed, including the potential for conflict. PMID- 11388326 TI - [Neural control of the circulation in normal and pathologic conditions]. AB - The concept of sympatho-vagal balance is particularly useful to explore some functional features of cardiovascular neural regulation. The variability of signals like RR interval or systolic arterial pressure, assessed with power spectrum analysis, has provided for the first time, simultaneously, markers of sympathetic and vagal modulation, and hence of their interaction. The relative power of low frequency (LF) oscillation quantifies sympathetic modulation, while the high frequency (HF) oscillation quantifies vagal modulation. LF and HF components are also detectable in the variability of muscle sympathetic nerve activity recorded in man. In numerous physiological and pathophysiological conditions these two oscillations undergo a reciprocal relationship similar to that characterizing sympatho-vagal balance. An increase in sympathetic modulation has been found during different physiological stimuli and in abnormal states such as recovery from myocardial infarction, essential arterial hypertension, and congestive heart failure in its early stage. A correct use of this methodology is prevented by a drastic decrease in variance, often accompanying pathophysiological reductions in responsiveness of target functions. In these cases the decrease in LF power yields an important negative prognostic value. PMID- 11388327 TI - [Heart rate variability]. AB - In 1981 a study on heart rate variability (HRV) and on the neural control of the heart was published in Science. Since then, the number of papers concerning this topic has increased exponentially. Some aspects of the relationship between HRV and the neural control of the cardiovascular system are now clear, for example: a decreased SDNN (a measure of variance) during a period of 24 hours is a well established independent negative prognostic factor after acute myocardial infarction and in chronic heart failure patients. Moreover, an increased sympathetic modulation elicited by standard laboratory maneuvers determines an increased power of the low frequency (LF) spectral component (0.03-0.15 Hz), expressed in normalized units. Nevertheless, the interpretation of the oscillatory components present in HRV, particularly in pathophysiological conditions, is far from being clear. The influence of several neurohumoral circuits can induce various changes in HRV according to different physiological or pathological conditions. A new approach to the study of HRV called "information domain" might help to obtain some more information about HRV. Furthermore, in the future, HRV analysis might be useful when deciding on the best pharmacological combination for the individual patient and for the probabilistic prediction of acute cardiac events. PMID- 11388328 TI - [Blood pressure variability]. AB - This review deals with a number of issues related to blood pressure variability. These include: historical aspects, with reference to the first pioneering observations; methodological aspects, focusing on the different methods for quantifying blood pressure variability; description of the characteristics of blood pressure variability over the 24 hours; mechanisms involved in determining the different magnitude of this phenomenon in different subjects, such as behavioral factors, central and reflex neural influences, humoral and mechanical factors; blood pressure variability as a probe to assess spontaneous baroreflex sensitivity; effects of aging and hypertension on blood pressure variability, with a discussion of the clinical relevance of this phenomenon in the prognostic evaluation of patients; effects of drugs on blood pressure variability. Finally methodological aspects related to the use of noninvasive ambulatory blood pressure monitoring in the assessment of blood pressure variability are discussed. PMID- 11388329 TI - [Baroreflex sensitivity]. AB - Arterial baroreceptors play an important role among the large number of physiological mechanisms governing the adjustment of cardiovascular system to several surrounding conditions. By baroreceptor stimulation, arterial pressure changes can modulate both sympathetical and vagal activity and, as a consequence, heart rate, contractility and vascular resistance. In the last years, many experimental and clinical observations have shown that ischemic heart disease and heart failure can change baroreceptor reflex sensitivity and cause excessive or inappropriate activity of the sympathetic system. Several methods have been developed to measure baroreceptor sensitivity by estimating the extent of change in heart rate following blood pressure oscillations being them spontaneous or brought about by application of pharmacological or mechanical stimuli. Under normal clinical conditions these measurements can be taken as the ability to activate a sympathetic answer (hypotension) or a parasympathetic one (hypertension), with the interplay of tonic vagal or sympathetic activity. The methodology most extensively used in the clinical setting relies on intravenous administration of phenylephrine, a pure alpha-agonist drug that activates arterial baroreceptors and leads to a reflex bradycardia, which can be measured as RR interval prolongation. Baroreflex sensitivity is quantified in ms of RR interval prolongation for each mmHg of arterial pressure increase. Compared to values obtained in normal subjects (average 15 ms/mmHg) baroreflex sensitivity is significantly depressed in post-infarction patients and in patients with heart failure. The application of a mechanical stimulus is carried out by means of a positive or negative pneumatic pressure through a collar around the neck. A decrease in neck chamber pressure, by stretching carotid receptors, is sensed as an arterial pressure increase and activates reflex bradycardia at the sinus node. Finally, the analysis of spontaneous oscillations of arterial pressure and heart rate can also provide information about baroreflex control of the cardiovascular system: indeed, even small physiological variations in arterial pressure can evoke a reflex heart rate response brought about by arterial baroreceptor. The potential clinical interest of these measurements (completely non-invasive) must be still studied in large populations to define both range of normality and prognostic significance. PMID- 11388330 TI - [Cardiopulmonary sensitivity and chemosensitivity]. AB - The autonomic control of the cardiovascular system plays an important role in maintaining the arterial pressure at the levels necessary for adequate tissue perfusion. In cardiovascular diseases, the impairment of the basic reflex mechanisms that are responsible for the moment-to-moment regulation could increase sympathetic activity and is correlated with an adverse outcome. The objective of the present review was to provide information about the methodological aspects exploring cardiopulmonary and chemoreceptor reflexes. Different techniques are available and all of them include assessment of reflexes through the activation or deactivation of either the cardiopulmonary baroreceptors or chemoreceptors. Intravenous saline load, head-down tilt, passive legs raising, head-out water immersion and the application of a lower body positive pressure are the principal methods utilized for activating cardiopulmonary baroreceptors; on the contrary deactivation could be achieved by acutely induced hypovolemia by furosemide or blood donation, inflation of a congestion cuff on the thighs or application of a negative pressure on the lower body. The transient exposure to a hypoxic or a hypercapnic gas mixture is frequently used to determine the peripheral and central chemoreflexes, respectively. The reflexes are quantified by the gain between output (i.e. heart rate, sympathetic activity, vascular resistance, ventilation) and input (oxygen saturation, end-tidal CO2 or changes in central venous pressure). One important limitation in assessing the cardiopulmonary baroreflex by using currently available techniques is that the involvement of the arterial baroreflex cannot be avoided. In addition, chemoreflexes cannot be interpreted unless the breathing rate is controlled. To date, several techniques are available for the quantification of cardiopulmonary baroreceptor and chemoreceptor reflexes and could provide new information on the abnormal autonomic mechanisms contributing to the pathophysiology of several cardiovascular diseases. PMID- 11388331 TI - [Tilt test and orthostatic intolerance: abnormalities in the neural sympathetic response to gravitational stimulus]. AB - In the present manuscript the different methodologies aimed at assessing the autonomic profile in humans during a gravitational stimulus have been described. In addition, strengths and drawbacks of the tilt test in relation to occasional orthostatic intolerance were addressed. Finally, different autonomic abnormalities underlying occasional and chronic orthostatic intolerance syndromes have been schematically highlighted. The direct recording of the neural sympathetic discharge from the peroneal nerve (MSNA), in spite of its invasive nature, still represents the recognized reference to quantify the changes in the sympathetic activity to the vessels attending postural modifications. The increase of plasma norepinephrine during a tilt test is achieved by both an increase in plasma spillover and a concomitant decrease in systemic clearance. Changes in the indices of cardiac sympathetic and vagal modulation may also be quantified during a tilt test by power spectrum analysis of RR interval variability. The spectral markers of cardiac autonomic control, if evaluated concomitantly with MSNA, may contribute to assess abnormalities in the regional distribution of the sympathetic activity to the heart and the vessels. The capability of the tilt test of reproducing a vasovagal event or of inducing "false positive responses" seems to be markedly affected by the age, thus suggesting that additional or different etiopathogenetic mechanisms might be involved in the loss of consciousness in older as compared to younger subjects. In subjects suffering from occasional or habitual neurally mediated syncope an increase or, respectively, a decrease in cardiac and vascular sympathetic modulation has been documented before the loss of consciousness. In patients with pure autonomic failure, a global dysautonomia affecting both the sympathetic and the vagal modulation to the heart, seems to be present. In chronic orthostatic intolerance, the most common form of dysautonomia of young women, an abnormal regional distribution of sympathetic activity has been hypothesized during up right posture. Indeed, during standing a blunted increase of sympathetic activity to the vessels is attended by a cardiac sympathetic overactivity leading to an exaggerated tachycardia. PMID- 11388332 TI - [Spectral analysis of muscle sympathetic nerve activity in man]. AB - Recent applications of frequency domain analysis to the variability of muscle sympathetic nerve activity (MSNA) have improved the comprehension of the relationship between cardiovascular oscillations and the autonomic nervous system. It has been observed that spectral analysis of MSNA is characterized by two major oscillatory components at low (LF) and high (HF) frequencies, similar to those detectable in the variability of cardiovascular signals. Pharmacological and non-pharmacological studies have shown that, at least within the physiological range, the two MSNA rhythms show a reciprocal behavior, similar to that already observed for cardiovascular oscillations. The oscillatory pattern of MSNA provides non-redundant but complementary information with respect to the time domain measures of MSNA (burst rate and amplitude) since it has been shown that completely different spectral profiles may be derived from various MSNA recordings. On the other hand, the latter are instead comparable in terms of mass activity. Due to the intrinsic characteristics of the signal, which represents the direct outflow of the central neural structures of the cardiovascular autonomic nervous system, this approach can be considered as a unique window over the central organization of excitatory and inhibitory neural mechanisms responsible for the genesis and the regulation of cardiovascular oscillations. PMID- 11388333 TI - [The Genetic Area of the ANMCO. Family history in modern cardiology: cardiomyopathies -- Part II]. AB - The family history plays an important role in the cardiomyopathy setting. Cardiomyopathies are defined as familial when at least two members of the family are proven as affected. Given that the definition of familial cardiomyopathy has to be evidence-based, the familial forms have to be identified and documented. Detailed family pedigrees are obtained by interviewing patients and relatives and examining all clinical and pathological reports. Then, the clinical non-invasive screening of relatives is proposed, and performed in all informed and consenting relatives. All patients diagnosed with cardiomyopathy are potentially affected by familial forms, until relatives are proven to be unaffected. A few exceptions could be for syndromic disorders for which the phenotypes provide certainty elements/signs analogous to those observed in the proband. Key points for family history interpretation are the phenotype at onset, the time of onset, the presence/absence of coronary risk factors (such as diabetes and hypertension) and concomitant diseases. Special attention has to be paid to neuromuscular disorders that represent a wide heterogeneous issue in which cardiac involvement (cardiomyopathy, arrhythmias and conduction defects) could be the first manifestation of the disease. Based on rigorous investigation, the information derived for each family will provide useful data for present and future management of the family members, and for future research in the field of cardiomyopathies. PMID- 11388334 TI - [Revision and update of guidelines on acute ischemic cardiopathy. Acute myocardial infarction]. PMID- 11388336 TI - [Pravastatin and the development of diabetes mellitus. Evidence for a protective treatment effect in the West of Scotland Coronary Prevention Study]. PMID- 11388335 TI - [Superiority of clopidogrel versus aspirin in patients with prior cardiac surgery]. PMID- 11388337 TI - [Endoluminal beta-radiation therapy for the prevention of coronary restenosis after balloon angioplasty. Localized intracoronary gamma-radiation therapy to inhibit the recurrence of restenosis after stenting]. PMID- 11388338 TI - [Physical activity and risk for cardiovascular events in diabetic women]. PMID- 11388339 TI - [Performance at a hemodynamics laboratory]. PMID- 11388340 TI - Combination therapy with cerivastatin and gemfibrozil causing rhabdomyolysis: is the interaction predictable? PMID- 11388341 TI - Atorvastatin-induced dermatomyositis. PMID- 11388342 TI - Neutropenia during treatment with amiodarone. PMID- 11388343 TI - Recurrent Staphylococcus aureus extracerebral infections complicating hereditary hemorrhagic telangiectasia (Osler-Rendu-Weber disease). PMID- 11388344 TI - Two sides of the warfarin issue: diligence versus potency. PMID- 11388345 TI - Ebola-Athens preemergence? PMID- 11388346 TI - What is the future of quinacrine sterilation? PMID- 11388347 TI - Smoking and follicle numbers. PMID- 11388348 TI - Pregnancy subsequent to uterine artery embolization. PMID- 11388349 TI - Tissue factor expression in sickle cell anemia. PMID- 11388350 TI - Development of the Brazilian Amazon. PMID- 11388351 TI - Predicting human longevity. PMID- 11388352 TI - Human origins and ancient human DNA. PMID- 11388353 TI - Special issue on the interface of balance disorders and anxiety: an introduction and overview. PMID- 11388354 TI - Psychological factors influencing recovery from balance disorders. AB - This article reviews evidence for three mechanisms whereby psychological factors may aggravate dizziness and retard recovery from balance disorders. Firstly, a common behavioral response to dizziness is to avoid activities and environments that provoke symptoms, yet such avoidance deprives the individual of the exposure necessary to promote psychological and neurophysiological adaptation. Secondly, anxiety arousal and hyperventilation may add to, amplify, and disinhibit the somatic symptoms induced by balance disorder. Thirdly, attention and cognitive load may influence the central processing of information required for the perception and control of orientation. The need to combine physiotherapy for dizziness with psychotherapy is discussed. PMID- 11388355 TI - Behavior therapy for vestibular rehabilitation. AB - Patients with panic disorder and patients with vestibular disorders often share symptomatology, such as dizziness, spatial disorientation, and anxiety in particular environments. Because of the similar clinical presentations, it is not always apparent whether these symptoms are due primarily to a vestibular disorder or to panic disorder. Depending on where and how these patients enter the medical system, their symptoms may be remedied by treatment from behavioral therapists or physical therapists trained in vestibular rehabilitation. Although vestibular rehabilitation developed independently of behavioral treatment for anxiety disorders, there are remarkable similarities in treatment conceptualization and implementation. For example, both use exposure procedures designed to produce habituation of dizziness and disorientation, as well as enhancing functional compensation. Furthermore, there appears to be a subset of individuals with panic disorder who also have vestibular pathology and thus, may benefit from both interventions. In this paper, similarities and differences in the clinical presentation, treatment goals, and specific interventions for patients with panic disorder or vestibular pathology is examined, and future implications are discussed. PMID- 11388356 TI - Vestibular rehabilitation for patients with agoraphobia and vestibular dysfunction: a pilot study. AB - This study examined whether physical therapy with vestibular rehabilitation exercises would benefit patients with agoraphobia and vestibular dysfunction. Nine patients went through a 2-week no-treatment baseline phase, a 4-week behavioral treatment phase focusing on self-directed exposure, and an 8-12-week vestibular rehabilitation phase (weekly sessions). On the main outcome measure, clinical global impressions (CGI) ratings of severity, behavioral treatment was accompanied by a reduction in severity (effect size d=0.8; P<.10). On the supplementary measures, the Hamilton Anxiety Scale (Hamilton-A) and the Chambless Mobility Inventory (MI), no significant improvements were noted. After vestibular rehabilitation therapy, further improvement occurred in CGI severity (d=0.65; two tailed P<.10), and significant improvements occurred in the supplementary measures. The physical therapist identified motion-induced dizziness and disturbances in balance in most patients. These improved with rehabilitation. Although the results can be attributed to other explanations, they are not inconsistent with the hypothesis that vestibular dysfunction maintains agoraphobic symptoms in some patients. PMID- 11388357 TI - Background and history of the interface between anxiety and vertigo. AB - The comorbidity of vertigo and anxiety has been an integral component of the medical literature since antiquity. In the works of Plato, the same terms were used in the context of vertigo, inebriation, height vertigo, disorientation, and mental confusion. In classical medicine, vertigo had the ambiguous status of being both a disease per se and a symptom of other diseases such as hypochondriacal melancholy. Further, two etiologies were described for vertigo: an origin in the head (brain) and an origin in the hypochondria (abdominal viscera). In the course of the development of a detailed neurologic taxonomy of vertigo in the latter half of the nineteenth century, a debate ensued whether agoraphobia was a form of vertigo or a distinct psychiatric condition. Elucidation of this forgotten debate, within its historical context, provides insights into the recent rediscovery of the balance-anxiety interface. PMID- 11388358 TI - Neurological bases for balance-anxiety links. AB - This review paper examines neurologic bases of links between balance control and anxiety based upon neural circuits that are shared by pathways that mediate autonomic control, vestibulo-autonomic interactions, and anxiety. The core of this circuitry is a parabrachial nucleus network, consisting of the parabrachial nucleus and its reciprocal relationships with the extended central amygdaloid nucleus, infralimbic cortex, and hypothalamus. Specifically, the parabrachial nucleus is a site of convergence of vestibular information processing and somatic and visceral sensory information processing in pathways that appear to be involved in avoidance conditioning, anxiety, and conditioned fear. Monoaminergic influences on these pathways are potential modulators of both effects of vigilance and anxiety on balance control and the development of anxiety and panic. This neurologic schema provides a unifying framework for investigating the neurologic bases for comorbidity of balance disorders and anxiety. PMID- 11388359 TI - Visual influences on balance. AB - This paper discusses the impact of vision on balance and orientation in patients with vestibular disorders and in anxiety patients with space and motion discomfort (SMD). When the vestibular system is impaired, vision has a greater influence on standing postural control, resulting in greater sway when individuals are presented with erroneous or conflicting visual cues. Studies have shown that individuals with other motion sensitivities, such as motion sickness, also tend to rely on vision for balance and do not disregard erroneous visual cues. Recently, patients with anxiety disorders that include SMD also have been shown to have increased postural sway in conflicting visual environments, similar to patients with vestibular disorders. Thus, while specific vestibular deficits are not always directly associated with SMD, data regarding the impact of vision on balance suggest that some patients with SMD may have an underlying balance disorder. PMID- 11388360 TI - A clinical taxonomy of dizziness and anxiety in the otoneurological setting. AB - Dizziness can be associated with otologic, neurologic, medical, and psychiatric conditions. This paper focuses on the interface between otologic and psychiatric conditions. Because dizziness often is situation specific, concepts of space and motion sensitivity (SMS), space and motion discomfort (SMD), and space and motion phobia (SMP) are needed to understand the interface. We present a framework involving several categories of interactions between balance and psychiatric disorders. The first category is that of dizziness caused by psychiatric disorder (psychiatric dizziness), including hyperventilation-induced dizziness during panic attacks. The second category involves chance cooccurrence of a psychiatric disorder and a balance disorder in the same patient. The third category involves problematic coping with balance symptoms (psychiatric overlay). The fourth category provides psychological explanations for the relationship between anxiety and balance disorders, including somatopsychic and psychosomatic relationships. The final category, neurological linkage, focuses on the overlap in the neurological circuitry involved in balance disorders and anxiety disorders. PMID- 11388361 TI - A conceptual structure and methodology for the systematic approach to the evaluation and treatment of patients with chronic dizziness. AB - The patient with chronic dizziness should never be labeled with psychogenic dizziness. Chronic does not mean psychogenic. Chronic means that health care has been unsuccessful. A systematic approach that yields a comprehensive formulation and rational treatment plan will increase the probability of a successful outcome and return to health. The four perspectives of diseases, life stories, dimensions, and behaviors provide a comprehensive yet flexible methodology for the evaluation of the patient in distress with chronic and disabling dizziness. The design of a comprehensive treatment plan involves the determination of each perspective's contribution to the patient's distress and to what relative degree. This process recognizes that the perspectives are distinct from one another but complementary in illuminating the various reasons for a patient's distress. The perspectives come together as the formulation of the patient's case and offer a recipe for treatment rather than just a list of ingredients such as bio, psycho, and social. PMID- 11388362 TI - Studies of cortical interactions over short periods of time during the search for verbal associations. AB - Interactions between cortical areas were studied during the search for verbal associations and reading of words. The functional anatomy and the sequence of involvement of cortical areas during the solution of these tasks have been described previously, in studies using positron emission tomography and multichannel recordings of evoked potentials combined with identification of the locations of dipole sources [8, 9, 19, 25]. Cortical interactions reflected in terms of the synchronization of EEG rhythms were studied by developing a method based on correlating curve wavelets, which allows the moments at which this synchronization occurs to be identified over short periods of time comparable with the speeds of individual thought operations (up to 100 msec). Three main stages were identified in the search for associations. During the first 200 msec after stimulus presentation, cortical connections were seen between the right and left frontal areas; at 200-500 msec, there were connections between the frontal and the temporal-parietal areas; finally, at 450-700 sec, there were connections between the left temporal and the right frontal-central-temporal areas. These results are in good agreement with data obtained previously using other methods and supplement them with mapping data on cortical connections. A number of differences in the mechanisms of information processing during the search for associations and reading were also identified. PMID- 11388363 TI - Vector representation of associative learning. AB - I. P. Pavlov has shown that conditioned reflexes are selective both with respect to conditioned stimuli and to conditioned reflexes elicited by those conditioned stimuli. At the neuronal level, selective aspects of conditional stimuli are based on detectors selectively tuned to the respective stimuli. The selective aspects of conditioned reflexes are due to command neurons representing specific unconditioned reflexes. It can be assumed that conditioned reflexes result from association between selective detectors and specific command neurons. The detectors activated by a conditional stimulus constitute a combination of excitations--a detector excitation vector. The detector excitation vector acts on a command neuron via a set of plastic synapses--a synaptic weight vector. Plastic synapses are modified in the process of learning, making the command neuron selectively tuned to a specific conditioned stimulus. The selective tuning of a particular command neuron to a specific excitation vector referred to a conditioned stimulus is the basis of associative learning. The probabilities of conditioned reflexes elicited by conditional and differential stimuli implicitly contain information concerning excitation vectors that encode the respective stimuli. The contribution of the vector code to associative learning was explored combining differential color conditioning with intracellular recording from color coding neurons. It is shown that colors in carps and monkeys are represented on a hypersphere in the four-dimensional space similar with human color space. The basis of the color space is constituted by red-green, blue-yellow, brightness, and darkness neurons. PMID- 11388364 TI - Expression of the c-fos gene in the mouse brain during the acquisition of defensive behavior habits. AB - Experimental data reported here provide evidence that marked defensive motivation in conditions of an unavoidable electrocutaneous stimulus is accompanied in mice by increases in the expression to the c-fos gene in the hippocampus and cerebral cortex. As reinforcement and learning progress and the animals become able to achieve a useful result--avoiding the electrocutaneous shock--expression of the c fos gene in these brain structures decreases. The level of expression of the c fos gene was found to be different in animals in which defensive behavior had different efficacies. Expression was higher in mice making large numbers of errors than in mice with few errors during acquisition of the defensive habit. PMID- 11388365 TI - The role of the cholinergic system of the sensorimotor cortex of the rat brain in controlling different types of movement. AB - The role of the cholinergic system of the sensorimotor cortex of the Wistar rat brain in controlling various types of movements was assessed by studying the effects of microinjections of carbachol and scopolamine into the representation area of the forelimb on the performance of two types of fore-limb food-procuring movements--with and without pressure on an obstacle--as well as on the animals' locomotion. These studies showed that administration of the cholinergic agonist carbachol (0.03-3 microg) leads to slowing of both types of procuring movements and acceleration of locomotor activity in an open field. Injections of the cholinergic antagonist scopolamine (0.3-3 microg) into the same area accelerated procuring movements, while the animals' locomotor activity remained unaltered. These data indicate that the cholinergic system of the sensorimotor cortex has different regulatory influences on movement activity (locomotion) and the performance of learned movements requiring forelimb muscle tone to be maintained for different periods of time (the usual rapid movements used for extracting food from a narrow horizontal tube versus slow movements with additional tactile and tonic components). PMID- 11388366 TI - Vibrissectomy during early ontogenesis in rats disturbs the functional properties of cortical projection neurons. PMID- 11388367 TI - Memory impairment in Parkinson's disease. AB - A total of 52 patients with Parkinson's disease (PD), 11 with presumptive Alzheimer's disease (AD), and 20 healthy subjects were studied; subjects were aged 55-74 years. Neurological symptoms were assessed quantitatively, and the state of higher mental processes were evaluated using the Luriya method. A number of memory tests were also used. These studies showed that PD was almost always accompanied by memory impairment exceeding the age norm. The major mechanism for memory impairment in PD without dementia was inadequate independent organization of memory-related activity at the memorizing and retrieval stages. In PD with dementia, there was also a primary impairment. Differences in memory impairments were found in PD with dementia (deeper derangements of involuntary memory and of information processing during memorizing). Impairments of consolidation of traces were more dependent on the age at onset of PD, while inadequacy of independent organization of memory-related activity was more dependent on disease duration. Most memory parameters in PD correlated with the severity of disturbances in gait and postural reflexes. It is suggested that memory impairment in PD is a manifestation of the major pathological process, which shows a number of differences from other neurogeriatric diseases. PMID- 11388368 TI - Autoantibodies to nerve growth factor in disorders of mental development in infants. AB - Studies were performed using 80 children aged 1-3 years with lesions to mental development. Patients were divided into three groups according to the severity and structure of lesions of mental development: those most strongly characterized by delayed speech development (group 1), those with delayed mental development due to organic CNS lesions (group 2), and those with abnormal mental development and high levels of psychopathological symptoms (group 3). There were significant increases (p < 0.001) in blood levels of autoantibodies to nerve growth factor (NGF) in children in each of these groups as compared with a control group (0.75 +/- 0.24 OD units). The levels of anti-NGF autoantibodies increased from group 1 to group 3 (0.95 +/- 0.24 OD units in group 1, 1.13 +/- 0.27 OD units in group 2. and 1.24 +/- 0.4 OD units in group 3). These correlations suggest that the level of anti-NGF autoantibodies can, taken in conjunction with a number of other parameters, be regarded as a potential molecular marker demonstrating abnormal development of the nervous system. PMID- 11388369 TI - A rare case of "mirror writing.". PMID- 11388370 TI - Memory impairment and deep brain structures. AB - Clinical and neuropsychological investigations were performed using the Luriya method in 141 patients with arteriovenous malformations (AVM). These included 27 patients with AVM in different parts of the caudate nucleus, 34 with AVM in the thalamus, 39 with AVM in the hippocampal formation, and 41 with AVM in the cingulate gyrus. A total of 102 patients underwent surgery. Patients with AVM in various locations showed common memory impairments as well as individual features of memory disturbances depending on which structures were affected. The common feature was the development of an amnesiac symptom complex resembling Korsakov's syndrome. These lesions developed only in patients with combined damage to deep structures (preoperatively in patients with ventricular hemorrhages); with the exception of patients with AVM of the caudate nucleus, memory impairments were modality-non-specific; all patients showed impairment of auditory-speech memory at the late phase and of visual memory in terms of indirect recall. Memory impairments characteristic of lesions to individual structures were the presence of functional asymmetry of memory defects in available of the caudate nucleus and thalamus (if only speech problems were present) and constant inclusions and contaminations in patients with AVM of the cingulate gyrus. It is suggested that certain aspects of memory function may have been duplicated in different structures during evolution and that each structure makes its own contribution to overall memory function. PMID- 11388371 TI - The angiotensin-converting enzyme gene as a possible risk or protective factor in Alzheimer's disease. PMID- 11388372 TI - The effects of aluminum ions on the phosphorylation of tubulin and microtubule proteins in the brain. PMID- 11388373 TI - The effects of an NDMA receptor antagonist on delayed visual differentiation in monkeys and rearrangements of neuron spike activity in the visual and prefrontal areas of the cortex. AB - The effects of perfusion of field 17 with the glutamate receptor antagonist 2 amino-5-phosphonovaleric acid (APV) on the characteristics of visual recognition and short-term memory were studied, along with the effects of APV on the responses of neurons in the visual and prefrontal areas of the cortex in rhesus macaques. In the test for delayed visual differentiation of stimuli of different colors, behavioral data were recorded simultaneously with multichannel recordings of the spike activity of single cells in cortical field 17 (directly within the microdialysis zone) and field 8. Multifactor dispersion analysis (ANOVA) showed that APV significantly worsened the behavioral characteristics in monkeys, with significant reductions in the duration of short-term storage of information (by factors of 2-4) and significant increases in the motor response times. These changes in cognitive characteristics induced by APV were accompanied by changes in the spike activity of neurons in the visual and prefrontal areas of the cortex during the sensory analysis and delay stages; changes in spike activity consisted of significant desynchronization. These results show that cognitive dysfunctions consisting of worsening of short-term remembering of information and increases in the duration of motor responses during exposure to APV may be caused by desynchronization of neuron activity in various areas of the cortex, these being involved in neuron ensembles responsible for the mechanisms of short-term memory, in which glutamatergic structures play an important role. PMID- 11388374 TI - Use of a simplified method of optical recording to identify foci of maximal neuron activity in the somatosensory cortex of white rats. AB - Eight mongrel white male rats were studied under urethane anesthesia, and neuron activity evoked by mechanical and/or electrical stimulation of the contralateral whiskers was recorded in the primary somatosensory cortex. Recordings were made using a digital USB chamber attached to the printer port of a Pentium 200MMX computer running standard programs. Optical images were obtained in the barrel field zone using a differential signal, i.e., the difference signal for cortex images in control and experimental animals. The results obtained here showed that subtraction of averaged sequences of frames yielded images consisting of spots reflecting the probable position of activated groups of neurons. The most effective stimulation consisted of natural low-frequency stimulation of the whiskers. The method can be used for preliminary mapping of cortical zones, as it provides for rapid and reproducible testing of the activity of neuron ensembles over large areas of the cortex. PMID- 11388375 TI - Neurophysiological correlates of delayed visual differentiation tasks in monkeys: the effects of the site of intracortical blockade of NMDA receptors. AB - A delayed visual differentiation task using stimuli of different colors was used in rhesus macaques to study the characteristics of visual recognition, short-term memory, and the responses of neurons recorded simultaneously in the visual and prefrontal areas of the cortex, along with their relationships with the site of intracortical (fields 17 or 46) perfusion of the glutamate receptor antagonist 2 amino-5-phosphonovaleric acid (APV). The behavioral characteristics and spike activity of individual cells in cortical fields 17 and 46 were recorded before and after perfusion with APV and after washing away of traces of APV. Multifactor dispersion analysis showed that the effect of APV in monkeys consisted of decreases in the probability of correct responses, leading to a decrease (two fold) in the duration of short-term storage of information and a significant increase in the motor reaction time. The probability of correct solutions depended on the site of APV perfusion in the cortex, while the motor response time was independent of the perfusion site. Perfusion of field 46 with APV, unlike perfusion of field 17, altered the spike activity only of neurons in the prefrontal cortex, while no significant changes were seen in the neuron activity of the visual cortex. The actions of APV were accompanied by significant desynchronization of neuron activity in these two areas as compared with the level of synchronization in normal conditions; after traces of APV were washed away, the extent of desynchronization decreased. The neurophysiological correlates of cognitive dysfunctions associated with degradation of visual recognition and short-term memory due to modification of glutamatergic structures by blockade of NMDA receptors are discussed. PMID- 11388377 TI - Studies of chemoreceptor perception in mollusks. AB - The formation and operation of the peripheral nervous system can be observed and studied in mollusks using as an example the primitive chemoreceptor organ, the osphradium, which is connected to the visceral arch of the CNS and analyzes the physicochemical properties of water in the mantle cavity. Nerve recordings showed that the osphradium is sensitive to excess osmotic pressure, sodium chloride, and amino acids. In addition, the osphradium responds to the quality of the water in which the animal is living. The osphradium of the pond snail retains its ancient multisensory function, uniting the perception of various chemical and physical stimuli. Patch clamp recordings at fixed potential or current were used to study membrane currents in identified ganglion and receptor cells, associated with increases in the concentrations of Na+ and L-aspartate in the solution bathing the osphradium. The influx current appears to be a sodium and/or calcium current, and is not blocked by tetraethylammonium, while the efflux current is a potassium current, as has been shown for the taste cells of vertebrates. PMID- 11388378 TI - No uniformity on informed consent. PMID- 11388376 TI - Pharmacological analysis of the subunit composition of the AMPA receptor in hippocampal neurons. AB - Experiments were performed on isolated neurons from hippocampal field CA1 and the dentate fascia to identify the subunit composition and distribution of splicing variants of AMPA receptor subunits. Currents evoked by the application of kainate were recorded using a whole-cell patch clamping method. The presence of GluR2 subunits in receptors was associated with a sharp reduction in the activity of the selective channel blocker IEM-1460. The composition of flip versions of subunits was assessed using cyclothiazide. AMPA receptors in the major cell types (pyramidal and granule cells) had low sensitivity to IEM-1460, while AMPA receptors of other cells (interneurons) had high or intermediate sensitivity. Cyclothiazide had strong potentiating effects on the main cell types in both structures as compared with interneurons. Thus, there is a correlation between the sensitivities of hippocampal neurons to IEM-1460 and cyclothiazide. The main cell types in both structures expressed large quantities of the GluR2 subunit in their AMPA receptors, with high levels of flip subunits, as compared with the other cell types, in which GluR2 subunits were virtually absent and the flop version predominated. This appears to reflect the functional features of different types of neurons. PMID- 11388379 TI - Early American oculoplastic surgery: the lectures of Dr. John Jeffries. PMID- 11388380 TI - Canthus-sparing ectropion repair. AB - PURPOSE: To describe a novel surgical technique for lower eyelid ectropion repair that avoids canthotomy and cantholysis and can be used in combination with external levator repair and/or in combination with blepharoplasty. METHODS: A retrospective analysis of lower eyelid procedures with the use of the canthus sparing technique between January 1, 1998, and December 31, 1999, was performed. The canthus-sparing approach was used in 198 eyelid procedures for the correction of lower eyelid ectropion. Seventy-four (37.4%) procedures involved the correction of lower eyelid ectropion alone and 25 (12.6%) procedures involved the correction of lower eyelid ectropion during upper eyelid small-incision external levator repair. In these cases, an incision was made lateral to the lateral canthus and a periosteal flap was created at the lateral orbital rim. The inferior crus of the lateral canthal tendon was then attached to this full thickness elevated periosteum. Twenty (10.1%) procedures involved the correction of ectropion during upper blepharoplasty and 79 (39.9%) procedures involved the correction of ectropion during combined upper eyelid ptosis repair and blepharoplasty. In these cases, the inferior crus of the lateral canthal tendon was attached to a periosteal flap created through the lateral portion of the blepharoplasty incision. RESULTS: The mean age of patients undergoing ectropion repair was 74.3+/-9.3 years (range, 42-93 years). The average duration of symptoms (most commonly tearing and/or ocular irritation) was 20+/-14 months (range, 3-84 months). Recurrences of lower eyelid ectropion or symptoms occurred in 4 (2%) eyelids. The average follow-up interval was 54+/-65 days (range, 3-330 days). CONCLUSIONS: The canthus-sparing approach to ectropion repair promotes a secure adhesion to the lateral orbital wall with minimal violation of normal anatomic structures and relations. It is time-efficient and reduces postoperative morbidity. PMID- 11388381 TI - Use of isobutyl cyanoacrylate tissue adhesive to stabilize external eyelid weights in temporary treatment of facial palsies. AB - PURPOSE: To describe a technique for stabilizing external eyelid load weights with isobutyl cyanoacrylate tissue adhesive for the temporary treatment of facial palsies. METHODS: This was a nonrandomized, prospective study of six consecutive patients with unilateral facial paralysis treated with external eyelid load weights stabilized with isobutyl cyanoacrylate tissue adhesive instead of adhesive tape. Follow-up assessment included corneal exposure, patient comfort, amount of artificial tear usage, and complications associated with the weights. RESULTS: Of the six patients studied, five had decreased corneal exposure, decreased artificial tear usage, and increased patient comfort with use of the weights. One patient had no improvement of symptoms. Weights were retained for a mean of 10.7 days. Two patients had difficulty with the weights; one was secondary to dermatochalasis and blepharoptosis obscuring his vision, and the other patient's weight fell off after 1 day. No weights were lost. CONCLUSIONS: The use of isobutyl cyanoacrylate tissue adhesive is a simple, quick, and inexpensive method for placement of external eyelid weights for temporary treatment of ocular exposure associated with facial paralysis and for determining the correct weight for implantation. Furthermore, isobutyl cyanoacrylate tissue adhesive used to stabilize external eyelid weights is better tolerated and lasts longer than the previously described fixation method with adhesive tape. PMID- 11388382 TI - Orbital fractures in children. AB - PURPOSE: To describe the demographics, etiologic factors, clinical presentations, and outcomes of orbital fractures in children. METHODS: This was a retrospective case series of 96 consecutive patients under 18 years of age with orbital fractures presenting to the Massachusetts Eye and Ear Infirmary, including both hospitalized and nonhospitalized patients. RESULTS: Orbital fractures in children were most frequently the result of sports, assault, or motor vehicle accident. The majority of patients did not require hospitalization and were treated as outpatients. The medial wall and floor of the orbit were the most frequent locations of fracture. Approximately half of the patients in this series required surgery, most often for entrapment. There were no cases of persistent diplopia in patients in whom surgery was performed or was not indicated. Associated ocular injuries were observed in half of the patients. CONCLUSIONS: In this series of hospitalized and nonhospitalized patients, orbital fractures in children had a location pattern similar to that most frequently observed in adult patients (floor and medial wall). Orbital fractures in children frequently require surgery. The high prevalence of ocular injury in children with orbital fractures emphasizes the need for a comprehensive ophthalmic evaluation. PMID- 11388383 TI - Endonasal dacryocystorhinostomy in the primary treatment of acute dacryocystitis with abscess formation. AB - PURPOSE: To determine whether endonasal dacryocystorhinostomy may constitute effective primary treatment of acute dacryocystitis with lacrimal sac abscess formation. METHODS: This was a retrospective review of a series of 24 patients with acute dacryocystitis and lacrimal sac abscess who underwent endonasal dacryocystorhinostomy as a primary procedure. Outcome measures included resolution of signs and symptoms of acute dacryocystitis as well as intranasal ostium patency as assessed by lacrimal irrigation. RESULTS: Pain was relieved in all patients within 3 days of surgery, and swelling resolved in all patients by 9 days after surgery. Ostium patency, as defined by the absence of epiphora, and free lacrimal irrigation was achieved in 20 (83%) of 24 patients, with follow-up of 27 to 59 months (mean, 40 months). Recurrent epiphora developed in four patients; recurrent dacryocystitis developed in none. CONCLUSIONS: Endonasal dacryocystorhinostomy may be a useful option in the treatment of acute dacryocystitis with abscess formation. PMID- 11388384 TI - Experience with 120 synthetic hydroxyapatite implants (FCI3). AB - PURPOSE: To assess the problems and/or complications associated with the use of a synthetic hydroxyapatite implant (FCI, Cedex, France). METHODS: The authors analyzed all of the problems and/or complications associated with the use of a third-generation synthetic hydroxyapatite implant (FCI3) in 120 patients by one surgeon over 4 years. The following data were recorded: age, type of surgery performed, size of implant used, peg system used, follow-up duration, time of pegging, problems and/or complications encountered, and treatment. RESULTS: Thirteen patients were lost to follow-up after 3 months, leaving 107 patients who were followed up from 4 to 48 months (average, 29 months). Discharge occurred in 21 (19.6%) patients, implant exposure in 3 (2.8%), socket discomfort in 2 (1.9%), trochleitis in 2 (1.9%), conjunctival thinning in 1 (0.93%), and pyogenic granuloma in 1 (0.93%). Peg problems occurred in 24 (35.2%) of 68 patients. Problems encountered with the peg were discharge in 10 (14.7%) patients, pyogenic granuloma in 9 (13.2%), conjunctiva overgrowing the peg in 4 (5.8%), hydroxyapatite exposure around the sleeve in 3 (4.4%), loose sleeve in 3 (4.4%), peg drilled on an angle in 1 (1.5%), implant infection in 1 (1.5%), and peg falling out in 1 (1.5%). CONCLUSIONS: The FCI3 synthetic hydroxyapatite is a less costly alternative form of hydroxyapatite currently in use in many parts of the world. Problems and complications encountered with its use are similar to those seen with the Bio-Eye Integrated Orbital Implants, Inc., San Diego, CA, U.S.A. The incidence of exposure associated with the synthetic hydroxyapatite implant is lower than several other reports on the Bio-Eye. The synthetic hydroxyapatite implant is slightly softer than the Bio-Eye and fractured under extreme pressure in one case. PMID- 11388385 TI - Metastatic renal cell carcinoma to the palpebral lobe of the lacrimal gland. AB - PURPOSE: To describe a clinicopathologic correlation of a metastatic renal cell carcinoma to lacrimal gland. METHODS: Case report. RESULTS: A 59-year-old man with a history of renal cell carcinoma had a hemorrhagic mass involving the palpebral lobe of the right lacrimal gland. Fundus examination disclosed two lesions with typical features of choroidal metastasis. The lacrimal gland mass was excised, and histopathologic examination revealed metastatic renal cell carcinoma. The patient was treated for systemic metastasis but required no further ocular treatment. CONCLUSIONS: Renal cell carcinoma can metastasize to the lacrimal gland, where it may appear as a hemorrhagic mass. PMID- 11388386 TI - From eyelid bumps to thyroid lumps: report of a MEN type IIb family and review of the literature. AB - PURPOSE: We present a two-generation family with multiple endocrine neoplasia (MEN) type IIb diagnosed by their ophthalmologists based on characteristic ophthalmic findings. METHODS: A family consisting of a 33-year-old female proband and her 8- and 7-year-old children had prominent corneal nerves; eyelid, lip, and tongue nodules; and a characteristic facies. A polymerase chain reaction-based genetic assay was obtained to detect the genetic mutation most commonly associated with MEN type IIb. Serum calcitonin and urine catecholamine studies were obtained. RESULTS: Molecular genetic studies detected in all 3 patients a mutation at codon 918 of the RET proto-oncogene known to be present in 95% of the cases of MEN type IIb. Serum calcitonin was elevated in the proband and her son. Urine catecholamine levels were elevated in the proband. Surgical treatment and histologic analysis confirmed pheochromocytoma and medullary thyroid carcinoma (MTC) in the proband. Surgical exploration revealed the MTC to be metastatic to the liver. CONCLUSIONS: This family demonstrates the characteristic findings of MEN type IIb: prominent corneal nerves in a clear stroma and multiple submucosal neuromas of the conjunctiva, eyelids, lips, and tongue. Ophthalmologists have a critical role to play in recognizing these signs, because the early diagnosis of medullary thyroid carcinoma and pheochromocytoma may be life saving. PMID- 11388387 TI - Giant cell angiofibroma of the nasolacrimal duct. AB - PURPOSE: To describe clinical and histologic features of the first case, to our knowledge, of giant cell angiofibroma located in the nasolacrimal duct region in a 28-year-old woman. METHODS: Interventional case report. A left nasolacrimal duct tumor was excised en bloc by lateral rhinotomy. Histopathologic examination was performed with the use of light microscopy. Immunohistochemical staining included S-100 protein, muscle-specific actin, desmin, myoglobin, vimentin, and CD34. RESULTS: The lesion was characterized by haphazardly arranged oval to spindled cells, a myxoid and collagenous stroma, multinucleated giant cells, prominent blood vessels, and pseudovascular spaces. Tumor cells were strongly positive for vimentin and CD34 and were negative for other antigens. After excision, there has been no recurrence over 4 years of follow-up. CONCLUSIONS: Originally described as an orbital tumor, giant cell angiofibroma also may occur in the nasolacrimal duct and lacrimal sac region. This mesenchymal neoplasm should be included in the differential diagnosis of lacrimal drainage system tumors. PMID- 11388388 TI - Orbital solitary fibrous tumor: radiographic and histopathologic correlations. AB - OBJECTIVE: To correlate the clinicopathologic and radiographic features characteristic of orbital solitary fibrous tumor (SFT). METHODS: The diagnostic features and clinical outcome of seven adults with orbital SFT are retrospectively outlined. Orbital imaging was performed by ultrasonography, computed tomography, or magnetic resonance imaging. Some cases were imaged by multiple modalities. Histopathologic examination of each tumor specimen included standard light and immunohistochemical stains. RESULTS: Heterogeneous internal composition was better appreciated on magnetic resonance imaging than on computed tomography. All cases undergoing magnetic resonance imaging showed T1 isointensity and T2 hypointensity relative to gray matter. Strong, generalized immunohistochemical reactivity to vimentin and CD34 validated the diagnosis of SFT and differentiated the specimens from other spindle cell neoplasms. After complete tumor resection, our patients remain tumor free with postoperative intervals of 15 to 45 months. CONCLUSIONS: Solitary fibrous tumor has now been reported in 26 orbits. No physical finding is pathognomonic, but several imaging traits are highly characteristic. Intralesional image heterogeneity and a predominantly low T2 signal intensity are distinctive of SFT. Complete tumor resection and immunohistologic specimen evaluation are emphasized. Clinicians should consider the diagnosis of SFT when confronted with an adult patient having an orbital soft tissue mass demonstrating the distinctive magnetic resonance imaging findings. PMID- 11388389 TI - Adrenocortical carcinoma metastatic to the orbit. AB - PURPOSE: To describe the clinical course and histopathologic features of a patient with adrenocortical carcinoma metastatic to the orbit. METHODS: Case report and literature review. RESULTS: A 24-year-old man first came to medical attention because of symptoms referable to a 4.47-kg, nonfunctioning carcinoma of the left adrenal cortex. Several metastases ensued, including a large tumor to the right superior lateral bony orbit with extension to the brain, temporalis fossa, and orbit proper. The tumor was resected with the use of a combined neurosurgical, ophthalmic, and craniofacial approach. The patient died of widespread metastatic disease 15 months after the orbital operation. CONCLUSIONS: Metastasis to the orbit from adrenocortical carcinoma is rare. Surgical resection is the treatment of choice, with adjunctive radiation therapy and chemotherapy in some cases. The prognosis is poor. PMID- 11388390 TI - Polyglactin mesh wrapping of hydroxyapatite implants. PMID- 11388391 TI - Recent concept of autoimmune-related pancreatitis. PMID- 11388392 TI - Prognostic value of p53 mutations in patients with locally advanced esophageal carcinoma treated with definitive chemoradiotherapy. AB - PURPOSE: A significant correlation has been found between p53 mutation and response to chemotherapy or radiotherapy. To determine the prognostic value of p53 mutation in patients with locally advanced esophageal carcinoma treated with definitive chemoradiotherapy, p53 mutation was analyzed using the biopsied specimens taken for diagnosis. METHODS: Concurrent chemoradiotherapy was performed for 40 patients with severe dysphagia caused by esophageal squamous cell carcinoma associated with T3 or T4 disease. Chemotherapy consisted of protracted infusion of 5-fluorouracil, combined with an infusion of cisplatinum. Radiation treatment of the mediastinum was administered concomitantly with chemotherapy. The p53 gene mutation was detected by fluorescence-based polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) methods. DNA sequences were determined for DNA fragments with shifted peaks by SSCP methods. RESULTS: Of the 40 patients, 15 had T3 disease and 25 had T4 disease; 11 patients had M1 lymph node (LYM) disease. Of the 40 patients, 13 (33%) achieved a complete response. The median survival time was 14 months, and the 2-year survival rate was 20%. Among the 40 tumor samples, p53 mutation was detected in 24 tumors (60%). The survival rate in the 24 patients with p53 mutation did not differ significantly from that in the 16 patients without p53 mutation. In contrast, the 15 patients with T3 disease survived longer than the 25 patients with T4 disease (P = 0.016); however, the survival rate in the 11 patients with M1 LYM disease did not differ significantly from that in the 29 patients without M1 LYM disease. CONCLUSION: Concurrent chemoradiotherapy is potentially curative for locally advanced esophageal carcinoma, but p53 genetic abnormality has no impact on prognosis. PMID- 11388393 TI - Ulcerative colitis in the elderly: clinical patterns and outcome in 51 Greek patients. AB - PURPOSE: In this study, the clinical characteristics and course of all patients with ulcerative colitis in whom diagnosis of the disease was made at or after the age of 60 (older group), were analyzed and compared with those of patients with ulcerative colitis in whom diagnosis of the disease was made before the age of 60 (younger group). METHODS: The older group consisted of 51 patients (28 men and 23 women; aged 64+/-3.1 years) and the younger group consisted of 362 patients (192 men and 170 women; aged 38.4+/-14.9 years). The mean follow-up times in the two groups were 9.3 and 12.6 years, respectively. RESULTS: No significant differences between the two groups were found as far as the extent of the disease, the severity of the initial episode, and the outcome of the first episode of ulcerative colitis were concerned. However, significantly fewer elderly patients were operated on for their disease compared with younger patients (6.25% vs 22.3%; P = 0.0268). Although some differences in the course of the disease between elderly and younger patients were observed, such as the number of exacerbations and recurrences and the number of patients who developed colorectal cancer, these differences did not reach statistical significance. CONCLUSION: It is concluded that ulcerative colitis in elderly Greek patients runs a rather similar course to that in younger patients. However, some unique characteristics observed in the elderly patients (lower rate of colectomy, absence of patients with colorectal cancer, and increased death rate) could be attributed either to truly different disease behavior in the elderly people or to factors directly related to their advanced age. PMID- 11388394 TI - Seroepidemiological study of hepatitis E virus infection in Japan using a newly developed antibody assay. AB - PURPOSE: A seroepidemiological study of hepatitis E virus (HEV) infection was conducted in Japan, where HEV infection is not considered endemic. METHODS: IgG and IgM class antibodies to HEV were measured with a newly developed enzyme linked immunosorbent assay in which recombinant virus-like particles were used as an antigen. A total of 1253 individuals (401 males and 852 females; age range, 6 89 years) were enrolled from two different areas: area 1 (n = 478), in which hepatitis C was endemic; and area 2 (n = 775), in which it was not endemic. RESULTS: The HEV antibody (IgG class) positive rate was 6.7% in area 1 and 4.6% in area 2. Similarly, the HAV antibody (IgG class) positive rates were 65.3% and 72.3%. The age- and sex-specific prevalence of both HAV and HEV antibodies was quite similar in the two areas, and the HAV antibody positive rate clearly increased with age in both males and females. On the other hand, the HEV antibody positive rate showed a slight tendency to increase with age in males, but not in females. None of the 32 individuals with the HEV antibody who were interviewed had a history of visiting countries in which hepatitis E was endemic. In both areas, the mean age, percentage of males, and HAV antibody positive rate were significantly higher in the group of individuals with the HEV antibody than in the group of those without it, according to conventional statistical analyses. Of the three factors age, male sex, presence of HAV antibody, and the area factor, only male sex was statistically significant (P < 0.001) on multivariate logistic regression analysis. Two (0.2%) of the total of 1253 individuals were positive for the IgM class antibody to HEV. CONCLUSIONS: Our results suggest the possibility that HEV infection is circulating in Japan at a low level. HEV infection was associated with male sex, but not with HAV infection. PMID- 11388395 TI - Characteristics of the cell proliferation profile of activated rat hepatic stellate cells in vitro in contrast to their fibrogenesis activity. AB - PURPOSE: Alterations in the kinetics of hepatic stellate cells (HSCs) after the cells are activated once have not been well documented. We investigated the characteristic profiles of cell proliferation of once-activated HSCs in contrast to the in fibrogenesis activity. METHODS: HSCs from male Wistar rats were submitted to primary culture for 14 days and to secondary culture for 7 days. The potential for cell proliferation was evaluated by the number of the cells in G2/M phase, based on flow cytometric analysis of the cell cycle. The fibrogenesis activity was assessed by Northern blot analysis of the expression of type I and type III procollagen mRNA. RESULTS: The number of HSCs in G2/M phase was maintained at a low level in primary culture after 6 days, while a significantly (P < 0.05) elevated number of HSCs in G2/M phase was observed on days 3 to 4. In secondary culture, the number of HSCs in G2/M phase was also consecutively maintained at a decreased level. By contrast, HSCs showed progressively increased type I and type III procollagen mRNA expression during the experimental periods of primary culture. CONCLUSIONS: These results clearly demonstrated consecutively decreased proliferative activity, evaluated by the potential for cell mitogenesis, in once-activated HSCs, in contrast to their progressively increased fibrogenesis activity. PMID- 11388396 TI - Sludge and stone formation in the gallbladder in bedridden elderly patients with cerebrovascular disease: influence of feeding method. AB - PURPOSE: The incidence of gallbladder sludge or gallstone formation in bedridden patients with cerebrovascular disease (CVD) remains obscure. The aim of this study was to determine the incidence, relationship to feeding method, and mechanisms of gallbladder sludge and gallstone formation in elderly patients with CVD. METHODS: Using ultrasonography, we determined the development of gallbladder sludge and gallstone over a 12-month period, the area of the gallbladder, the gallbladder contractile response to cerulein, and fasting levels of plasma cholecystokinin (CCK) in 40 bedridden elderly patients with CVD. The patients were divided into three groups based on the feeding method: oral ingestion (OI), nasogastric feeding (NF), and total parenteral nutrition (TPN). RESULTS: Gallbladder sludge and gallstone were not observed in any of the 14 OI patients, but occurred in 6 and 1 of the 11 NF patients, and in 14 and 3 of the 15 TPN patients, respectively. Fasting gallbladder areas were significantly larger in the TPN group than in the other two groups. The TPN group showed a marked decrease in cerulein-induced gallbladder contractility. Fasting plasma CCK levels were lower in the TPN group than in the OI group. CONCLUSIONS: Our results indicate that elderly patients with CVD confined to bed over long periods are not necessarily at risk of gallbladder sludge or gallstone formation, and the development of these features may be associated with the feeding method. The predisposition of CVD patients on TPN to gallbladder disease is probably caused by failure of gallbladder contraction, resulting from insufficient secretion of CCK and impaired sensitivity of the gallbladder to CCK. PMID- 11388397 TI - Pneumatosis intestinalis and hepatic portal venous gas caused by mesenteric ischemia in an aged person. AB - An 82-year-old woman complaining of abdominal pain and vomiting was admitted to our emergency department. Abdominal X-ray, ultrasonography, and computed tomography showed hepatic portal venous gas, as well as pneumatosis intestinalis. We first suspected superior mesenteric arterial thrombosis. However, her physical findings, including computed tomography scanning and laboratory data, did not support the presence of bowel necrosis. The gas disappeared after 1 day. After the 12th day, she had recovered with conservative therapy, and she was discharged on the 41st day. Many reports indicate that hepatic portal venous gas is often associated with bowel necrosis, and urgent operation is recommended in such instances. In this patient, total colonoscopy on the 7th day revealed longitudinal redness, suggesting mesenteric ischemia. Thus, we speculate that this is a rare case of mesenteric ischemia without bowel necrosis associated with both pneumatosis intestinalis and hepatic portal venous gas. PMID- 11388398 TI - Budd-Chiari syndrome and extrahepatic portal obstruction associated with congenital antithrombin III deficiency. AB - We report a patient with Budd-Chiari syndrome (BCS) and extrahepatic portal obstruction (EHO) associated with congenital antithrombin (AT) III deficiency. A 35-year-old man was admitted to Nishi Kobe Medical Center for evaluation of abnormal intrahepatic veins. By various imaging modalities, BCS and EHO were diagnosed. Laboratory data revealed parallel decreases in activity and antigen concentration of AT III despite normal liver function. Taken together, the etiology of both BCS and EHO was considered to be thrombosis, associated with congenital AT III deficiency. Two years after beginning warfarin therapy, the patient has no symptoms and his liver function remains normal. Anticoagulant therapy is considered useful for preventing progression of the disease. PMID- 11388399 TI - Hepatocellular carcinoma depicted as hypoattenuation on CT hepatic arteriography (CTA) and hyperattenuation on CT during arterial portography (CTAP). AB - We report a 68-year-old man with three nodules of hepatocellular carcinoma (HCC) in a cirrhotic liver; the largest nodule was 3.0cm in diameter. The nodules showed hypoattenuation on computed tomography (CT) hepatic arteriography (CTA) and hyperattenuation on CT during arterial portography (CTAP), indicating that the dominant vascularity of the HCC nodules may have been the portal vein. A biopsy specimen obtained from the nodules showed well differentiated HCC (Edmondson-Steiner grade I). The imaging findings of the nodules on both CTA and CTAP are unusual, in spite of the rather large size, so this seemed suggestive of the hemodynamic properties of relatively large nodules of well differentiated HCC. PMID- 11388400 TI - HBV-related fulminant hepatic failure: successful intensive medical therapy in a candidate for liver transplantation. AB - Fulminant hepatic failure (FHF) usually has a fatal prognosis without liver transplantation. We describe the case of a woman who developed FHF, and was evaluated as a candidate for liver transplantation, but who was cured without transplantation through intensive medical care that included glucagon-insulin therapy, methylprednisolone pulse therapy, interferon beta and lamivudine administration, cyclosporine administration, and high-volume hemodiafiltration and plasma exchange. In a patient with FHF who is a candidate for liver transplantation but for whom the transplantation cannot be performed for some reason, intensive medical therapy, including regeneration-promoting therapy, immunosuppressive therapy, antiviral therapy, and vigorous hepatic support, should be carried out. PMID- 11388401 TI - Are the clinical features of ulcerative colitis different in the elderly? PMID- 11388402 TI - Gallbladder sludge and stone formation vary with the pathophysiological condition. PMID- 11388403 TI - Amiloride-sensitive sodium currents in identified taste cells of the frog. AB - Sodium ions occurring in food are thought to be detected, at least in part, through specific amiloride-sensitive, sodium channels (ASSCs) localized in taste receptor cells. Cells within taste buds are morphologically heterogeneous, and include taste receptor cells and other cells that could perform a support or even transduction role. It is not known whether subsets of the taste bud cells express ASSCs, and whether the properties of these channels are similar. By applying the patch-clamp technique to morphologically distinct cells, the supporting wing cells, isolated from the frog taste disk, I have found functional ASSCs that are moderately sensitive to amiloride (Ki 3-4 microM) and which are distinctly lower in affinity for amiloride than reported frog taste receptor cells (Ki 0.2 microM). These results support the hypotheses of the existence of distinct, functional ASSCs in different cell morphotypes, at least in frog taste organs. PMID- 11388404 TI - Antagonism of ORLI receptor produces an algesic effect in the rat formalin test. AB - The authors investigated the role of endogenously released nociceptin (also known as orphanin FQ) spinal and supraspinal nociceptive transmission during the rat formalin test by examining the effect of intrathecal and intracerebroventricular injection of J-113397, a non-peptidyl ORL1 receptor selective antagonist. When J 113397 was injected intrathecally or intracerebroventricularly 10 min before the formalin injection, it enhanced the agitation behavior induced by paw formalin injection. This suggested that paw formalin injection induced nociceptin release in the spinal cord and the supraspinal brain sites, that this endogenously released nociceptin produced an analgesic effect and that J-113397 antagonized this analgesic effect of nociceptin and produced an algesic effect in the rat formalin test. PMID- 11388405 TI - A comparison of the induction of immortalized endothelial cell impermeability by astrocytes. AB - The suitability of various commercially available endothelial cell lines in studies of astrocytic/endothelial cell interactions was assessed. The endothelial like cell line ECV304 was compared with T24/83, Eahy929, and b.End5 and rat cerebral endothelial cells in their ability, when co-cultured with rat (C6) glioma cells, to form a transendothelial electrical resistance (TEER), an indicator of tight junction formation which is an important property of the blood brain barrier. As reported previously, the basal TEER of ECV304 cell monolayers was significantly enhanced upon co-culture, an effect reproduced by human 1321N1 astrocytes and primary rat astrocytes. T24/83 cells formed a patchy, gapped monolayer, which produced a poor basal TEER with little in the way of an increase upon co-culture. Similarly, all the other cell monolayers analysed demonstrated poor TEERs that were only moderately increased upon co-culture. These data confirm that while no endothelial cell line with ideal features is available, ECV304 cells remain an appropriate choice especially for studies of astrocyte/endothelial cell interactions. PMID- 11388406 TI - Reorganisation of the sensorimotor cortex after early focal brain lesion: a functional MRI study in monozygotic twins. AB - Sensorimotor cortical reorganization after early brain lesions was studied by means of fMRI in two pairs of monozygotic twins, in each of which one member had a focal brain injury. This offered a unique opportunity to reduce the wide intersubject variability of the controls often found in similar studies. Activation images were acquired during a motor task (sequential opposition finger movements) and a sensory task (passive brushing of palm and fingers). During the tasks with the recovered hand, constant findings in the lesioned subjects were the activation of the undamaged areas adjacent to lesion site and the activation of the ipsilateral sensorimotor cortex. Bilateral activation of the primary sensorimotor cortex was never observed in the healthy co-twin controls. PMID- 11388407 TI - Observation-execution matching system for speech: a magnetic stimulation study. AB - Observation of limb movements in human subjects resulted in increased motor evoked potential (MEP) amplitude elicited by magnetic stimulation of motor cortex in the muscles involved in that movement, suggesting that an observation execution matching (OEM) system exists in humans. We investigated whether the OEM system is activated by speech gestures presented in the visual and auditory modalities. We found that visual observation of speech movement enhanced MEP amplitude specifically in muscles involved in production of the observed speech. In contrast, listening to the sound did not produce MEP enhancement. The findings suggest that the OEM system may be modality specific. It may be involved in action recognition in the visual modality, but is not responsible for perception of simple items of sound. PMID- 11388408 TI - Characterization of a brain-related serine protease, neurosin (human kaillikrein 6), in human cerebrospinal fluid. AB - Neurosin (also known as zyme or protease M) is a trypsin-like serine protease dominantly expressed in the human brain. According to the official nomenclature, this gene is now designated as human kallikrein 6 (KLK6) and the protein is designated hK6. To investigate the metabolism of neurosin in human brain, neurosin contained in the human cerebrospinal fluid (CSF) was analyzed. Neurosin was detected in the all CSFs tested by Western blot analysis using an anti neurosin monoclonal antibody. We purified neurosin from CSF (CSF-neurosin) using an immunoaffinity chromatography and an anion-exchange chromatography. SDS-PAGE revealed that the purified protein has a relative mol. mass (Mr) of 25,000 Da. The observed sequence of the N-terminal amino acids, Glu-Glu-Gln-Asn-Lys, of the purified CSF-neurosin was identical to the sequence of N-terminal of the pro enzyme form, which is presumed to have no enzyme activity. CSF-neurosin neither showed any enzyme activity to Boc-Phe-Ser-Arg-4-methylcoumaryl-7-amide, which is known to be degraded by the mature neurosin, nor cleaved gelatin. To confirm that the major portion of CSF-neurosin is present in the pro-enzyme form, Western blot analysis using antibodies specific to the pro- or mature enzyme was carried out. The antibody against the mature neurosin fragment did not react with CSF neurosin. Only the antibody against the pro-enzyme fragment detected CSF neurosin. Thus, our results suggest that neurosin is present as an inactive pro enzyme in the human CSF. PMID- 11388409 TI - Tetrodotoxin-sensitive enhancement of inhibition in CA1 pyramidal neurones by nicotine. AB - Nicotine modulates excitatory and inhibitory transmission in the hippocampus by acting on receptors located on various cellular compartments. We report that nicotine, applied for 5-10 min at concentrations similar to those found during smoking (0.5-5 M), resulted in all CA1 pyramidal neurones in a marked, phasic and tonic increase in the frequency and amplitude of spontaneous inhibitory currents. This effect was fully prevented by pre-incubation with the sodium channel blocker tetrodotoxin and was partially inhibited by the two nicotinic receptor antagonists methyllicaconitine (MLA) and dihydro-beta-erythroidine (DHbetaE). We conclude that, under conditions found during smoking, nicotine enhances inhibitory transmission, an effect exclusively mediated through an enhancement of the firing rate of interneurones, without changes in spontaneous quantal release of GABA. PMID- 11388410 TI - Malonate-induced cortico-motoneuron death is attenuated by NT-4, but not by BDNF or NT-3. AB - Neurotrophins are promising candidates to slow the progression of amyotrophic lateral sclerosis (ALS), a neurodegenerative disease in which spinal and cortical motoneurons selectively degenerate. In a long-term in vitro model, malonate induced toxicity and cell death of motoneurons have been demonstrated. Here we studied the neuroprotective effect of BDNF, NT-3, and NT-4 on the cell death of cortical motoneurons in an organotypic culture model after chronic mitochondrial inhibition with malonate. Our data show that NT-4 completely prevents malonate induced toxicity, whereas BDNF or NT-3 had no neuroprotective effect. In clinical trials for ALS, predominantly focussed on the survival of spinal motoneurons, BDNF has already been tested with disappointing results; our results suggest that NT-4 may be a better neurotrophin to prevent motoneuron loss. PMID- 11388411 TI - EAAT1 and EAAT2 immunoreactivity in transgenic mice with a G93A mutant SOD1 gene. AB - Immunocytochemical and quantitative analyses were used to correlate the localisation of excitatory amino acid transporter proteins EAAT1, EAAT2 with time in spinal motoneurones of presymptomatic and symptomatic mice with the G93A mutant SOD1 gene. Specimens from age-matched non-transgenic wild-type mice served as controls. EAAT1 and EAAT2 immunoreactivity was well-preserved in the gray matter in both controls and transgenic mice at all ages, and there was no difference in the expression of EAAT1 and EAAT2 immunoreactivity between controls and transgenic mice. These findings suggest that EAAT1 and EAAT2 may not play a pivotal role in the degeneration of motoneurons in this animal model. PMID- 11388412 TI - Lethal comatose mutation in Drosophila reveals possible role for NSF in neurogenesis. AB - NSF is a cytosolic ATPase implicated in a variety of cellular functions including synaptic vesicle exocytosis. Here we report a lethal mutation in the Drosophila homolog of NSF (dNSF1). Lethality staging and rescue experiments with the wild type dNSF1 transgene show that NSF1 is critically required during early larval stages and during late pupariation. Lethality in larval stages is associated with defects in neurogenesis as evidenced by an overall reduction in synapse size and synapse branching. Moreover, escaper adults, though showing abnormal seizure-like paralytic behavior, are normal in terms of synaptic transmission as assayed by electroretinograms. Taken together, these data indicate a role for NSF in neural growth and branching in addition to its documented role in synaptic transmission. PMID- 11388413 TI - Effects of cannabinoids in Krox-24 targeted mice. AB - Krox-24 is an immediate early gene encoding a zinc-finger transcription factor implicated in several adaptive responses, and its induction by cannabinoids has been reported. We used mice targeted in the Krox-24 gene to specifically dissect the role of this protein in the acute and chronic central actions of cannabinoids. We report here on the ability of cannabinoids to activate G proteins and to inhibit adenylyl cyclase, and to elicit behavioral responses in wild-type and mutant mice. The behavioral parameters and the biochemical correlates of abstinence after delta9-THC withdrawal were evaluated. We show that Krox-24 is not involved in the acute analgesic effects of delta9-THC and in the SR precipitated delta9-THC withdrawal syndrome. PMID- 11388414 TI - Immunocytochemical analysis of cholinergic amacrine cells in the tiger salamander retina. AB - Cholinergic amacrine cells in the tiger salamander retina were observed for the first time by using antibodies against choline acetyltransferase (ChAT). ChAT immunoreactive cells were present in the inner nuclear layer (INL) and in the ganglion cell layer (GCL), and the somas of the former population (average diameter = 15.13 microm) were slightly smaller than those of the latter population (average diameter = 16.42 microm). The processes of these cells form two distinct narrow bands in the inner plexiform layer (IPL), one located near 0.2 inner plexiform units (IU) and the other near 0.65-0.7 IU. Soma size, cell density and spatial distribution of ChAT-positive cells were quantitatively analyzed. Our results suggest that cholinergic amacrine cells in the salamander retina are very similar to their counter parts in other species, and they can be used as a model system for studying cholinergic functions in the visual system. PMID- 11388416 TI - Differentiating ERAN and MMN: an ERP study. AB - In the present study, the early right-anterior negativity (ERAN) elicited by harmonically inappropriate chords during listening to music was compared to the frequency mismatch negativity (MMN) and the abstract-feature MMN. Results revealed that the amplitude of the ERAN, in contrast to the MMN, is specifically dependent on the degree of harmonic appropriateness. Thus, the ERAN is correlated with the cognitive processing of complex rule-based information, i.e. with the application of music-syntactic rules. Moreover, results showed that the ERAN, compared to the abstract-feature MMN, had both a longer latency, and a larger amplitude. The combined findings indicate that ERAN and MMN reflect different mechanisms of pre-attentive irregularity detection, and that, although both components have several features in common, the ERAN does not easily fit into the classical MMN framework. The present ERPs thus provide evidence for a differentiation of cognitive processes underlying the fast and pre-attentive processing of auditory information. PMID- 11388415 TI - Survival and plasticity of basal forebrain cholinergic systems in mice transgenic for presenilin-1 and amyloid precursor protein mutant genes. AB - The basalo-cortical cholinergic system was characterized in mice expressing mutant human genes for presenilin-1 (PS1), amyloid precursor protein (APP), and combined PS/APP. Dual immunocytochemistry for ChAT and A beta revealed swollen cholinergic processes within cortical plaques in both APP and PS/APP brains by 12 months, suggesting aberrant sprouting or redistribution of cholinergic processes in response to amyloid deposition. At 8 months, cortical and subcortical ChAT activity was normal (PS/APP) or elevated (PS, APP frontal cortex), while cholinergic cell counts (nBM/SI) and receptor binding were unchanged. ChAT mRNA was up-regulated in the nBM/SI of all three transgenic lines at 8 months. The data indicate that the basal forebrain cholinergic system does not degenerate in mice expressing AD-related transgenes, even in mice with extreme amyloid load. The PMID- 11388418 TI - Optokinetic reflex dysfunction in multiple sclerosis. AB - The aim of this study was to further investigate optokinetic reflex function in multiple sclerosis. Gaze-holding in darkness, optokinetic nystagmus, optokinetic afternystagmus and latency to circularvection were measured using electro oculography and a rotating optokinetic drum. Gaze-holding was not significantly different between the multiple sclerosis and control groups; however, four of 23 multiple sclerosis patients exhibited eccentric gaze-evoked nystagmus. There were no significant differences in either optokinetic nystagmus frequency or latency to circularvection. However, optokinetic nystagmus slow phase velocity during rise time and amplitude during beat time were significantly reduced in the multiple sclerosis group (p < 0.05 and p < 0.0001, respectively). The time constant of optokinetic afternystagmus was also significantly reduced in the multiple sclerosis group (p < 0.005). These results indicate that optokinetic nystagmus and optokinetic afternystagmus are significantly impaired multiple sclerosis. PMID- 11388417 TI - Induction of cytokine expression in the brain macrophages/amoeboid microglia of the fetal rat exposed to a teratogen. AB - Using in situ hybridization, RT-PCR, lectin histochemistry and immunohistochemistry, this study examined the time course expression and cellular localization of various cytokines including tumor necrosis factor-alpha (TNF alpha), interleukins (IL-1, IL-6) and transforming growth factor-beta (TGF-beta) in fetal rat brain after a maternal injection of the teratogen cyclophosphamide (CP). Eight hours after CP injection, there was a marked increase in brain macrophages (BM)/amoeboid microglia (AM) in different areas of the fetal brain as determined by lectin histochemistry. Concomitant to this was the induction in mRNA level of TNF-alpha, which was progressively increased with time. TGF-beta mRNA was undetectable until 24 h had elapsed. Expression of IL-1 and IL-6 was undetectable at all stages. In situ hybridization combined with immunohistochemistry has shown the localization of TNF-alpha in BM/AM and neurons. Present results suggest that both TNF-alpha and TGF-beta are involved in the progression of neural damage in the fetal brain induced by the teratogen. PMID- 11388419 TI - Expression of HGF and cMet in the peripheral nervous system of adult rats following sciatic nerve injury. AB - Hepatocyte growth factor (HGF) exhibits neurotrophic properties on different types of neuron, including motor, sensory and parasympathetic neurons. We demonstrate that sciatic nerve ligation induces an increase of the HGF receptor, c-met, mRNA in the distal segment of the sciatic nerve to the ligation site and a delayed elevation in the proximal segment. Immunohistochemical analysis revealed co-localization of cMet and GFAP and indicates that Schwann cells express cMet in the sciatic nerve after injury. HGF mRNA was detected in the spinal cord and DRG, and nerve injury did not alter the expression. These data demonstrate that the expression of HGF and cMet in the peripheral nervous system shows the unique pattern of regulation following nerve injury. PMID- 11388420 TI - An fMRI study of anticipation and learning of smooth pursuit eye movements in humans. AB - We studied the neural substrate of anticipation and learning of smooth pursuit eye movements in humans using fMRI. Both predictable and non-predictable eye movements, compared to baseline, activated a common network previously associated with oculomotor function. The temporal dynamics of activity in a subset of these areas suggested a strong correlation with type of condition. Specifically, differential decreases in activity were seen in dorsolateral prefrontal cortex and the intraparietal sulcus during the predictable condition. During the non predictable condition the same areas exhibited evidence of high levels of activity that further increased throughout the condition. In contrast, differential increases associated with the predictable condition were seen in anterior cingulate and preSMA cortex regions. These changes in activity mirror the time course of the short-term learning of eye movements seen behaviourally, and are congruent with learning-related changes that have been reported for other motor paradigms. PMID- 11388421 TI - Functional MRI with reduced susceptibility artifact: high-resolution mapping of episodic memory encoding. AB - Visual episodic memory encoding was investigated using echoplanar magnetic resonance imaging at 2.0 x 2.0 mm2 resolution and 1.0 mm section thickness, which allows for functional mapping of hippocampal, parahippocampal, and ventral occipital regions with reduced magnetic susceptibility artifact. The memory task was based on 54 image pairs each consisting of a complex visual scene and the face of one of six different photographers. A second group of subjects viewed the same set of images without memory instruction as well as a reversing checkerboard. Apart from visual activation in occipital cortical areas, episodic memory encoding revealed consistent activation in the parahippocampal gyrus but not in the hippocampus proper. This finding was most prominently evidenced in sagittal maps covering the right hippocampal formation. Mean activated volumes were 432 +/- 293 microl and 259 +/- 179 microl for intentional memory encoding and non-instructed viewing, respectively. In contrast, the checkerboard paradigm elicited pure visual activation without parahippocampal involvement. PMID- 11388422 TI - A standing Na+ conductance in rat carotid body type I cells. AB - Substitution of extracellular Na+ with N-methyl D-glucamine caused marked hyperpolarisation in rat isolated carotid body type I cells, suggesting the presence of a standing Na+ conductance. Choline substitution produced smaller hyperpolarisations, whilst Li+ was virtually without effect. This Na+ conductance was not blocked by amiloride, tetrodotoxin, Zn2+ or Gd3+ and did not arise from electrogenic Na-glucose co-transport, since substitution of glucose with sucrose could not mimic the effects of Na+ substitution. Hypoxia and acidosis did not modify the tonic Na+ influx. Our results suggest that Na+ influx provides a constant depolarising influence on type I cells which acts to shift membrane potential beyond that required for initiation of neurosecretion, an essential step in carotid body chemotransduction. PMID- 11388423 TI - Adrenal gland SNAP-25 expression is altered in thyroid hormone receptor knock-out mice. AB - SNAP-25 is a protein in neurons and neuroendocrine cells, which is involved, together with syntaxin and VAMP, in neurotransmitter release and neurite outgrowth. Since the thyroid hormone receptors TR alpha and TR beta are essential for nervous system development, their possible role in regulating the expression of these vesicle trafficking proteins was examined by analysing SNAP-25 levels in TR alpha and TR beta knock-out mice. Immunoblotting and RT-PCR showed that SNAP 25 levels are increased in the adrenal gland, but not in cerebellum, in knock-out mice, while syntaxin-1 and VAMP-2 are unaffected in either tissue. Treatment of the pheochromocytoma-derived cell line PC12 with the thyroid hormone L-3,5,3' triiodothyronine (T3) decreased SNAP-25 expression. Together, these data suggest that thyroid hormones exert a negative regulatory effect on SNAP-25 in adrenal medullary neuroendocrine cells. PMID- 11388424 TI - Reward-related neuronal activities in basal ganglia of domestic chicks. AB - We aimed to reveal what is coded in the basal ganglia of domestic chicks. In the water-reinforced 'go' task, chicks learned to peck selectively at a colored bead in order to obtain a drop of water. Out of 38 units obtained, seven showed excitatory activities specifically during the reward period. In the food reinforced go/no-go task, chicks learned to discriminate two colors to obtain mash food after a delay period. They also learned to ignore another color, which was not associated with a reward. Out of 27 units obtained, four showed excitatory activities during the cue period, specifically when a food reward was anticipated. LPO neurons may code qualities of the obtained rewards, and also chick's anticipation of the forthcoming rewards. PMID- 11388425 TI - Kir subfamily in frog retina: specific spatial distribution of Kir 6.1 in glial (Muller) cells. AB - We show by immunocytochemistry in frog retina that most members of the Kir subfamily are expressed in specific neuronal compartments. However, Kir 6.1, the pore-forming subunit of K(ATP) channels, is expressed exclusively in glial Muller cells. Muller cell endfeet display strong Kir 6.1 immunolabel throughout the retina, whereas the somata are labeled only in the retinal periphery. This spatial pattern is similar to that of Kir 4.1, of the ratio of inward to outward K+ currents, and of spermine/spermidine immunoreactivity. We suggest that the co expression of Kir 4.1 and Kir 6.1 subunits may enable the cells to maintain their high K+ conductance and hyperpolarized membrane potentials both at high ATP levels (Kir 4.1) and during ATP deficiency (Kir 6.1). PMID- 11388426 TI - Tuning in caudal fastigial nucleus units during natural and galvanic labyrinth stimulation. AB - Neurons of the caudal fastigial nucleus were investigated by means of single unit recordings. Natural vestibular stimuli were applied as well as galvanic labyrinth polarization. One-third of the neurons showed a convergence of vertical and horizontal canals. More than 80% of the neurons responded to polarization of both the ipsilateral and contralateral canals (binaural responders). Most neurons had a limited response range. Two classes of neurons could be distinguished: up to 1 Hz responders and up to 10 Hz responders. In addition a group of fastigial cells showed a tuning within a small range of frequencies (sharp-tuning responders). PMID- 11388427 TI - Reactive oxygen species mediate A beta(25-35)-induced activation of BV-2 microglia. AB - Microglial activation induced by beta-amyloid (A beta) is an important cellular response in the pathogenesis of Alzheimer's disease (AD). In this study, we show that reactive oxygen species (ROS) play a role as signaling molecules for the activation of NF-kappaB and induction of IL-1beta mRNA expression in A beta(25 35)-treated murine microglia BV-2 cells. ROS scavengers such catalase and superoxide dismutase (SOD) mimetics obviously reduced activation of NF-kappaB and the elevated level of IL-1beta transcripts induced by A beta(25-35). In addition, the A beta(25-35)-induced NF-kappaB activation and IL-1beta expression were suppressed by blockers of the ROS generating enzymes such as NADPH oxidase, cyclooxygenase, and lipoxygenase. These data suggest that ROS mediate A beta induced microglial activation. PMID- 11388428 TI - Pseudoginsenoside-F11 attenuates morphine-induced signalling in Chinese hamster ovary-mu cells. AB - Pseudoginsenoside-F11 (PF11), an ocotillol type saponin isolated from Panax quinquefolium L., has been shown to antagonize the behavioral actions of morphine. Biochemical experiments revealed that PF11 could inhibit diprenorphine (DIP) binding with an IC50 of approximately 6.1 microM and reduced the binding potency of morphine in Chinese hamster ovary (CHO)-mu cells. Furthermore, PF11 significantly attenuated morphine-stimulated [35S]GTPgammaS binding in a dose dependent manner, and strongly decreased the efficacy of morphine to inhibit intracellular cAMP production. In addition, PF11 pretreatment could also significantly inhibit naloxone induced cAMP overshoot in the morphine-pretreated cells. However, PF11 per se had no effect on either [35S]GTPgammaS binding or intracellular cAMP accumulation. These data suggested that PF11 antagonized the morphine stimulated opioid receptor signalling directly at the cellular level. PMID- 11388429 TI - Astroglial function of schizophrenic brain: a study using lobotomized brain. AB - The neurodevelopmental hypothesis is now being recognized as one of the most useful hypothesis for schizophrenia, and by using it, abnormalities in protein associated with neuron growth or neuronal migration have been reported. From neuron-glia interrelations in the neural development, it is important to study the function of astroglia in the schizophrenic brain. In this study, we examined the neuropathological reaction of astroglia using lobotomized brains, and a significant decrease of astroglia after artificial histological damage was observed in schizophrenic brains. We speculated that this may be due to the latent vulnerability of the dynamic function of astroglia in schizophrenia. Astroglia plays a guidance role on migration and if astroglia has latent vulnerability, we speculate that younger neurons may not sufficiently migrate during development. In further investigation of the neurodevelopmental hypothesis of schizophrenia, it will be necessary to examine the function of astroglia. PMID- 11388430 TI - Mouse model for morningness/eveningness. AB - Human morning/evening preferences has recently been reported to be associated with polymorphism of the 3' flanking region of the Clock gene, which was the first identified mammalian circadian clock gene. We recorded body temperature, spontaneous activity, electroencephalogram and electromyogram for 48 h in mice with Jcl:ICR genetic background and homozygous for the Clock mutation (Cl/Cl on Jcl:ICR). In both wild-type and Cl/Cl on Jcl:ICR, body temperature, activity, wake and sleep were completely entrained to LD cycle. However, phases of the rhythm for body temperature, activity and wake duration in the Cl/Cl on Jcl:ICR were about 2 h delayed in comparison with the wild-type. This study has provided further evidence on the close relationship between human morning/evening preference and the molecular basis of circadian clock system, and has suggested that Cl/Cl on Jcl:ICR is useful for an animal model for human morning/evening preference. PMID- 11388432 TI - Interhemispheric transfer time in a patient with a partial lesion of the corpus callosum. AB - The interhemispheric transfer time (ITT) of basic visuo-motor integration was investigated in a patient who had a lesion of the corpus callosum that spared the splenium and rostrum. Overall, 4291 simple reaction times were collected during unimanual responses to tachistoscopically presented lateralized simple visual stimuli at 4 degrees, 6 degrees and 10 degrees. Despite retaining some abilities that typically require the integration of information between hemispheres (e.g. haptic naming, tachistoscopic lateralized consonant reading) the patient performed similarly to completely callosotomized patients in a basic visuo-motor ITT task (overall 25.5 ms) at any eccentricity. These findings suggest that specific callosal channels mediate the basic visuo-motor ITT and these do not include the rostrum and/or the splenium of the corpus callosum. PMID- 11388431 TI - Intrathecal administration of endothelin-1 receptor antagonist ameliorates autoimmune encephalomyelitis in Lewis rats. AB - The role of endothelin-1 (ET-1) in the development of experimental autoimmune encephalomyelitis (EAE) was studied by the blocking the action of ET-1 with a receptor antagonist, BQ-123. Intrathecal administration of BQ-123 significantly ameliorated EAE progression at the peak stage of EAE (p<0.05). By immunohistochemistry, ED-1-positive macrophages in EAE lesions were identified as major producers of ET-1, whereas the immunoreactivity of ET-1 on brain cells, such as astrocytes, was dramatically increased in accordance with the progression of EAE. This study points to a putative pro-1nflammatory role for ET-1 in the pathogenesis of EAE. One possible application for the ET-1 receptor antagonist might be helpful in the therapy of autoimmune neurological disorders. PMID- 11388433 TI - Autoreceptor preference of dopamine D2 receptor agonists correlates with preferential coupling to cyclic AMP. AB - Dopamine autoreceptors control the synaptic release and turnover of dopamine. Some dopamine agonists display a preference for modulation of autoreceptor functions rather than postsynaptic-driven behaviors. However, the nature of this apparent selectivity is still elusive. To investigate this property, we have used an heterologous expression system in which D2S receptors are coupled to both inhibition of cyclic AMP levels and stimulation of inositol triphosphate production. We show that D2-like receptor agonists display distinct potencies on these two second messenger pathways. Moreover, a strong correlation is observed between the potency of agonists to interact with adenylate cyclase and their potency to modulate autoreceptor functions. Such a correlation does not show up with the phospholipase C pathway. This suggests that autoreceptor preference of D2-like receptor agonists may be driven by a preferential interaction with a second messenger system. PMID- 11388434 TI - Magnetic resonance imaging predicts neuropathology from soman-mediated seizures in the rodent. AB - Intoxication by the organophosphate compound soman causes prolonged seizures that lead to neuropathology in the brain. This MRI-based study describes the temporal and spatial evolution of brain pathology that follows soman-induced convulsions. We observed significant decreases in apparent diffusion coefficients (ADC; 23% below control) of the hippocampus and thalamus by 12 h after soman treatment. The ADC then returned to near normal values in all regions at 24 h but declined again during the next 7 days. These data suggest that the initial cellular degradation may be resolved but is ultimately followed by regional cellular remodeling. T2 relaxation values declined significantly at 12 h (37% decrease) returning to near normal values by 24 h. These data lend detail to the model suggesting that injured tissues experience an edematous influx that is resolved by 24 h. The imaging data was fully supported by histopathological comparisons where moderate cell loss and swelling within the hippocampus and piriform cortex was observed. This is the first report providing excellenttemporal and spatial resolution of emerging soman-mediated, seizure-induced neuropathology using MRI with histological correlation. PMID- 11388435 TI - Phase-specific modulation of cortical motor output during movement observation. AB - The effects of different phases of an observed movement on the modulation of cortical motor output were studied by means of transcranial magnetic stimulation (TMS). A video-clip of a reaching-grasping action was shown and single TMS pulses were delivered during its passive observation. Times of cortical stimulation were related to the phases of the shown movement, locking them to the appearance of specific kinematic landmarks. The amplitude of the motor evoked potentials (MEPs) induced by TMS in the first dorsal interosseus (FDI) muscle was modulated by the amount of the observed finger aperture. The presence of such an effect is consistent with the notion of a mirror neuron system in premotor areas that couples action execution and action observation also in terms of temporal coding. PMID- 11388436 TI - Calcium channel immunoreactivity in the salamander retina. AB - This study reports the distribution of the alpha1D and alpha1E calcium channel subunits in the neotenous tiger salamander retina based on immunohistochemical techniques. Confocal and light microscopy were used to localize staining with fluorescently tagged antibodies to alpha1D and alpha1E in cross-sectional and flatmount preparations of retina. Alpha1D-immunoreactivity (alpha1D-IR) was localized to the inner and outer plexiform layers (IPL and OPL, respectively), ganglion cell layer (GCL), and optic fiber layer. Alpha1E-IR was found predominantly in the IPL, with scattered, weak representation in the OPL. Alpha1E IR was not detected in the GCL or fiber layer. These findings suggest that different alpha1 calcium channel proteins have distinctive distributions in retina, which may reflect their unique and different roles in retinal processing and homeostasis. PMID- 11388437 TI - Human standing balance is affected by exposure to pulsed ELF magnetic fields: light intensity-dependent effects. AB - There is evidence in animals that behavioral and physiological responses to static and extremely low frequency magnetic fields (ELFMF) is affected by the presence of light during magnetic field exposures. Here we report that the effect of a specific pulsed ELFMF (PEMF) on human standing balance is modulated by light intensity during exposure. Under a low light condition (0.12 W/m2), nine healthy human volunteers stood on a 3D forceplate, throughout four 2 min exposures (eyes open/eyes close, sham/PEMF of 200 +/- 1 microTpk, order randomized). There was a significant increase in standing movement during PEMF exposure during eyes closed. In a second experiment on 26 normal subjects exposed to the identical protocol, but at greater light intensities (0.51 W/m2), a significant but opposite effect was observed. PMID- 11388438 TI - Rolipram reduces excitotoxic neuronal damage. AB - Proinflammatory cytokines are supposed to be involved in the pathophysiology of neuronal damage following excitotoxic lesions. We examined the effect of rolipram, a TNF-alpha-inhibitor, on excitotoxic neuronal damage. Quinolinic acid (240 nmol in 1 microl) was injected stereotactically into the striatum of male Wistar rats. Four groups of QA rats were treated i.p. with solvent, MK-801 (4 mg/kg) or rolipram (0.3 mg/kg) which was started either 6 or 24 h after QA injection and continued with daily applications for 14 days. QA injection induced neuronal damage which affected 93% of the striatal area. MK-801 reduced this damage to 12% of the striatal area. Treatment with rolipram when started at 6 h after QA injection resulted in neuronal damage amounting to 60%; the result after starting at 24 h was not different from solvent (91%). The present results demonstrate that rolipram reduces neuronal damage induced by intrastriatal QA application. PMID- 11388439 TI - Various glutathione S-transferase isoforms in the rat cochlea. AB - The localization of three glutathione S-transferase (GST) isoforms in the rat cochlea was examined using specific antibodies against each isoform. GST immunoreactivities were found in particular parts of the cochlea, including the intermediate cells and the basal cells of the stria vascularis and various types of fibrocytes in the spiral ligament. The different cell types showed varying combinations of GST isoforms. The GST immunopositive cells identified in the present study may play a central role in the metabolism and inactivation of endogenous and exogenous ototoxic compounds. The specific arrangements also indicated a possible contribution to the detoxification process in the form of a blood-labyrinth barrier. PMID- 11388440 TI - Fast vigilance decrement in closed head injury patients as reflected by the mismatch negativity (MMN). AB - Event-related potentials (ERPs) were measured from 24 chronic closed head injury (CHI) patients and 18 age- and education-matched controls. The oddball paradigm was applied while subjects were watching a silent movie. The standard (p=0.8) sound of 75 ms duration had a basic frequency of 500 Hz with harmonic partials of 1000 Hz and 1500 Hz, whereas these frequencies for the pitch deviant were each 10% higher. The frequencies of the duration deviant matched with those of the standard but was 25 ms in duration. The MMN (mismatch negativity), generated by the brain's automatic auditory change-detector mechanism, was elicited by both deviants. No significant differences in the MMN latency or amplitude for either pitch or duration deviants were found between the groups. However, the MMN amplitude for the pitch deviant decreased in the patient group during the experiment considerably faster than in controls, suggesting a faster vigilance decrement in the patients. PMID- 11388441 TI - Loss of vesicular zinc and appearance of perikaryal zinc after seizures induced by pilocarpine. AB - The condition of status epilepticus induced by systemic administration of kainic acid (KA) causes an apparent translocation of vesicular zinc from presynaptic boutons into postsynaptic neurons. The accumulation of zinc in the somata has been identified as a contributing cause of neuronal injury. We show here that another form of status epilepticus, induced by administration of the muscarinic agonist pilocarpine, produces changes in zinc that are essentially the same as those produced by the kainic acid-induced seizures. Moreover, neurons that develop zinc staining after pilocarpine seizures are the same that shown degenerative changes. This result suggests that the loss of zinc from presynaptic boutons and the appearance of zinc in postsynaptic somata may both occur in seizures per se, regardless of etiology. PMID- 11388442 TI - Are dyslexics' visual deficits limited to measures of dorsal stream function? AB - We tested the hypothesis that the differences in performance between developmental dyslexics and controls on visual tasks are specific for the detection of dynamic stimuli. We found that dyslexics were less sensitive than controls to coherent motion in dynamic random dot displays. However, their sensitivity to control measures of static visual form coherence was not significantly different from that of controls. This dissociation of dyslexics' performance on measures that are suggested to tap the sensitivity of different extrastriate visual areas provides evidence for an impairment specific to the detection of dynamic properties of global stimuli, perhaps resulting from selective deficits in dorsal stream functions. PMID- 11388443 TI - Task-dependent early latency (30-60 ms) visual processing of human faces and other objects. AB - Electrophysiological responses to previously seen faces reportedly differ from those to novel faces at shorter latencies than generally associated with complex visual analysis. It is unclear, however, whether such observations are unique to faces, and which stages of visual processing they reflect. MEG was used in 21 normal adults to record neural responses to images of faces, other objects and abstract patterns presented individually as part of a classification task and in sequential pairs as part of an image comparison task. The amplitudes of the short latency responses (30-60 ms) to the first image in pairs of faces were significantly greater than the responses to both the second faces and the individual face images. These early responses were recorded over predominantly right hemisphere parietal and occipito-temporal cortical regions including areas that, at longer latencies, have been associated with face specific activity. The differences in the responses within pairs were less for non-face objects and absent for abstract geometrical patterns. No early neuronal activity was observed in the classification task. The results indicate the existence of early latency neural networks that are sensitive to both stimulus type and task and are strongly activated by faces. PMID- 11388444 TI - Muscarinic dependence of nucleus basalis induced conditioned receptive field plasticity. AB - Receptive field (RF) plasticity in primary auditory cortex of adult animals, specifically selective increased response to a tonal conditioned stimulus (CS) relative to other frequencies, can be induced both by behavioral conditioning and by pairing a tone with stimulation of the nucleus basalis (NB). This study determined whether cortical muscarinic receptors are necessary for NB-induced RF plasticity. Single units in layers II-IV were studied in Urethane anesthetized adult rats. The cortex was perfused with saline or saline+atropine sulfate. Conditioning, 30 trials of pairing a tone with NB stimulation, produced a significant CS-specific response increase (n=8). Local atropine blocked NB induced RF plasticity, actually resulting in CS-specific response decrease (n=6). Therefore, NB-induced RF plasticity requires engagement of muscarinic receptors in auditory cortex. PMID- 11388445 TI - Functional recovery after brain lesion--contralateral neuromodulation: an fMRI study. AB - Behavioral recovery takes place even after permanent damage to the entire brain region normally controlling sensorimotor hind limb function in the rat. In our study, 2 weeks after full behavioral recovery from an experimental unilateral permanent brain damage, the topographic representation of the previous paretic hindlimb was investigated by fMRI. The analysis showed that during electrical stimulation of the previously paretic hindlimb, two normally inactive brain regions were now being activated. One region was the non-damaged contralateral sensori-motor cortex and the other region was located lateral to the lesion. These results suggest that behavioral recovery can be explained by functional reorganization and neuromodulation of the brain. PMID- 11388446 TI - From stem cells towards neural layers: a lesson from re-aggregated embryonic retinal cells. AB - Cells from dissociated embryonic avian retinae have the capacity to re-aggregate in rotation culture and form cellular spheres reconstituting a complete arrangement of all retinal layers. This exquisite phenomenon is based upon in vitro proliferation of multipotent precursor stem cells and spatial organization of their differentiating descendants. The addition of soluble factors from cultured retinal pigmented epithelial (RPE) or radial glial cells is essential to revert inside-out spheres (rosetted retinal spheres) into correctly laminated outside-out spheres (stratified spheres). Such complete restoration of a laminated brain tissue by cell re-aggregation has been achieved only for the embryonic avian retina, but not the mammalian retina, nor for other brain parts. This review summarises the history of the re-aggregation approach, presents avian retinal re-aggregate models, and analyses roles of the RPE and Muller cells for successful retinal tissue regeneration. It is predicted that these results will become biomedically relevant, as stem cell biology will soon open ways to produce large amounts of human retinal precursors. PMID- 11388447 TI - Small-angle X-ray scattering analysis of stearic acid modified lipase. AB - Stearic acid modified lipase (from Rhizopus japonicus) exhibited remarkable interesterification activity in n-hexane, but crude native lipase did not. The structure of the fatty acid modified lipase had not been analyzed until now. We analyzed the modified lipase by small-angle X-ray scattering (SAXS) measurements in order to clarify the structure. SAXS measurements showed that the modified lipase consisted of a lipid lamellar structure and implied that the lipase was incorporated into the lamellar structure of stearic acid. The long spacings in the lamellar structures of the modified lipase and stearic acid were measured. PMID- 11388448 TI - Effects of medium-chain fatty acids on intracellular calcium levels and the cytoskeleton in human intestinal (Caco-2) cell monolayers. AB - The effects of medium-chain fatty acids (MCFA) on intracellular calcium (Ca2+) levels and actin filaments in the Caco-2 monolayer were investigated. A site dependent increase in intracellular Ca2+ levels caused by decanoic acid (C10) at 13 mM was observed by confocal laser scanning microscopy. The area in which the intracellular Ca2+ levels was increased was measured by image analysis, and increased to 11% of the total area of the monolayer within 1 minute. This was maintained for 5 minutes, and decreased thereafter. The other MCFAs did not significantly increase the intracellular Ca2+ levels. Obvious morphological changes of actin filaments were induced by only C10 among C8-C14. The area in which actin filaments were depleted was also quantified, and the increase in area became significant after 40 minutes. The area of the actin-depleted spot corresponded to the area occupied by 5 to 10 cells as well as that in which the intracellular Ca2+ level was increased. The effectiveness of only C10 suggested that the mechanism of the absorption enhancement by C10 would be different from that by the other MCFAs, or that C10 has some additional physiological functions although the mechanism of the enhancement is the same as for the other MCFAs. PMID- 11388449 TI - The role of serine-246 in cytochrome P450eryF-catalyzed hydroxylation of 6 deoxyerythronolide B. AB - A strongly conserved threonine residue in the I-helix of cytochrome P450 enzymes participates in a proton delivery system for binding and cleavage of dioxygen molecules. 6-Deoxyerythronolide B hydroxylase (P450eryF) is unusual in that the conserved threonine residue is replaced by alanine in this enzyme. On the basis of the crystal structures of substrate-bound P450eryF, it has been proposed that the C-5 hydroxyl group of the substrate and serine-246 of the enzyme form hydrogen bonds with water molecules 519 and 564, respectively. This hydrogen bonding network constitutes the proton delivery system whereby P450eryF maintains its catalytic activity in the absence of a threonine hydroxyl group in the conserved position. To further assess the role in the proton delivery system of hydroxyl groups around the active site, three mutant forms of P450eryF (A245S, S246A, and A245S/S246A) were constructed and characterized. In each case, decreased catalytic activity and increased uncoupling could be correlated with changes in the hydrogen bonding environment. These results suggest that Ser-246 does indeed indirectly participate in the proton shuttling pathway, and also strongly support our previous hypothesis that the C-5 hydroxyl group of the substrate participates in the acid-catalyzed dioxygen bond cleavage reaction. PMID- 11388450 TI - Reaction kinetics and modeling of the enzyme-catalyzed production of lactosucrose using beta-fructofuranosidase from Arthrobacter sp. K-1. AB - Lactosucrose synthesis from sucrose and lactose was carried out by using beta fructofuranosidase from Arthrobacter sp. K-1. The transfructosylation mechanism was found to be of an ordered bi-bi type in which sucrose was bound first to the enzyme and lactosucrose was released last. Hydrolysis side-reaction experiments indicated that the reactions were uncompetitively inhibited by glucose and lactose, while no inhibition by fructose was apparent. The overall reaction rates were formulated. The reaction rate constants, equilibrium constant, and dissociation and Michaelis constants were determined at 35 degrees C and 50 degrees C by fitting the experimental concentration changes with the calculated values by a nonlinear least-square method. The average relative derivation for the concentrations was 9.67%. The kinetic parameters were also calculated for 43 degrees C and 60 degrees C by assuming the Arrhenius law, and the course of reaction was predicted. The obtained reaction rate equations well represented the concentration changes during the experiment at all temperatures. PMID- 11388451 TI - Molecular cloning and characterization of the fructooligosaccharide-producing beta-fructofuranosidase gene from Aspergillus niger ATCC 20611. AB - The fopA gene encoding a fructooligosaccharide-producing beta-fructofuranosidase was isolated from Aspergillus niger ATCC 20611. The primary structure deduced from the nucleotide sequence showed considerable similarity to those of two other beta-fructofuranosidases from A. niger, but the fopA gene product had several amino acid insertions and an extra C-terminal polypeptide consisting of 38 amino acids that could not be found in the two others. We could successfully express the fopA gene in S. cerevisiae and the fopA gene product obtained from the culture supernatant of the S. cerevisiae transformant had similar characteristics to the beta-fructofuranosidase purified from A. niger ATCC 20611. However, we could not detect any beta-fructofuranosidase activity in either the culture supernatant or cell lysate when the C-terminal truncated fopA gene product by 38 amino acids was used to transform S. cerevisiae. In western analysis of those samples, there was no protein product that is cross-reacted with anti-beta fructofuranosidase antibody. These results suggested that the C-terminal region of the fopA gene product consisting of 38 amino acids was essential for the enzyme production. PMID- 11388452 TI - Effects of brewer's yeast cell wall on constipation and defecation in experimentally constipated rats. AB - Brewer's yeast cell wall (BYC) was tested on constipated male Sprague-Dawley rats that had been induced by loperamide (2 mg/kg of body weight). The preventive effect of BYC on constipation was examined and compared with that of a non-fiber diet (NF) as the control. The dose-response of BYC and the effect on defecation by constipated experimental rats were also compared with the characteristics of cellulose diet (CE) group which served as a control. Defecation was observed to be greater by the rats fed with BYC than by those fed with NF or CE. The fecal water content and level of volatile fatty acids (VFA) in the cecal contents were likewise higher in the rats fed with BYC. These results indicate that the administration of BYC was effective for improving defecation and other parameters related to defecation. These favorable effects of BYC supplemented to the diet are attributed to the fermentation ability, water holding capacity and swelling force in the large intestine. PMID- 11388453 TI - Fibrinolytic and antithrombotic protease from Spirodela polyrhiza. AB - A fibrinolytic protease was purified from a Chinese herb (Spirodela polyrhiza). The protease has a molecular mass of 145 kDa and 70 kDa in gel filtration and SDS polyacrlamide gel electrophoresis (PAGE), respectively, implying it is a dimer. Its optimum pH was 4.5-5.0. The enzyme was stable below 42 degrees C and after lyophilization. The enzyme activity was inhibited significantly by leupeptin and aprotinin. The protease hydrolyzed not only fibrin but also fibrinogen, cleaving Aalpha and Bbeta without affecting the gamma chain of fibrinogen. It preferentially cleaved the peptide bond of Arg or Lys of synthetic substrates (P1 position). The enzyme had an anticoagulating activity measured with activated partial thromboplastin time (APTT), thrombin time (TT), and prothrombin time (PT) tests. It delayed APTT, TT, and PT two times at the concentration of 36, 39, and 128 nM, respectively and this was drastically reduced after heat treatment. PMID- 11388454 TI - ESR imaging on a solid-tumor-bearing mouse using spin-labeled dextran. AB - Imaging of a tumor with ESR was tried using two different types of spin probes, a low molecular weight spin probe, CPROXYL, and a polymer spin probe, TEMPO-DX. Spin probes were administered to a mouse bearing a solid tumor that was a transplanted Ehrlich's ascites carcinoma in the back, using two methods, conventional intraperitoneal injection and continuous intravenous injection with a micro-feeder. First, the accumulation of the probe was examined by X-band ESR. CPROXYL, which was administered to a mouse intraperitoneally, was exclusively retained in urine, showing that it was rapidly excreted into the bladder, while TEMPO-DX was absorbed from the peritoneal cavity with difficulty to the vessel. Using continuous intravenous injection, CPROXYL was also rapidly excreted, but it was confirmed that TEMPO-DX concentrated in tumor tissue because it has a long half-life in vivo. In addition, measurement of ESR imaging was done to measure the distribution of spin probes with continuous intravenous injection. The strongest spot of CPROXYL was observed on ESR images, showing the accumulation at the bladder, while the spot of TEMPO-DX was observed in the solid tumor of the back of the mouse. These results suggest that TEMPO-DX could stay much longer than a low molecular weight spin probe in vivo and concentrate at the tumor. TEMPO-DX may be useful for developing specific ESR imaging agents for tumor. PMID- 11388455 TI - S-Adenosyl-L-methionine-dependent O-methylation of 2-hydroxy-3-alkylpyrazine in wine grapes: a putative final step of methoxypyrazine biosynthesis. AB - The final step of 2-methoxy-3-alkylpyrazine (MP) biosynthesis has been presumed to involve O-methylation of 2-hydroxy-3-alkylpyarzine (HP), although this reaction has never been demonstrated in organisms. We detected 2-hydroxy-3 isobutylpyrazine (IBHP) and 2-hydroxy-3-isopropylpyrazine (IPHP) in unripe grapes, and S-adenosyl-L-methionine-dependent O-methyltransferase (OMT) activity toward HP in crude extracts from young shoots and unripe grapes that accumulate MP at different levels. The levels of HP in the berries and stems were estimated by using 2-hydroxy-3-sec-butylpyrazine as an internal standard. The OMT activity observed in the crude extracts from young shoot and berries was extremely low, but no MP-decomposing activity was detected in the solutions. The levels of HP and OMT activity were closely related with the level of MP in the grapes. These results indicate that the predicted final step of MP biosynthesis exists in wine grapes. PMID- 11388456 TI - Thermostable chitosanase from Bacillus sp. strain CK4: its purification, characterization, and reaction patterns. AB - A thermostable chitosanase, purified 156-fold to homogeneity in an overall yield of 12.4%, has a molecular weight of about 29,000 +/- 2,000, and is composed of monomer. The enzyme degraded soluble chitosan, colloidal chitosan, and glycol chitosan, but did not degrade chitin or other beta-linked polymers. The enzyme activity was increased about 2.5-fold by the addition of 10 mM Co2+ and 1.4-fold by Mn2+. However, Cu2+ ion strongly inhibited the enzyme. Optimum temperature and pH were 60 degrees C and 6.5, respectively. The enzyme was stable after heat treatment at 80 degrees C for 30 min or 70 degrees C for 60 min and fairly stable in protein denaturants as well. Chitosan was hydrolyzed to (GlcN)4 as a major product, by incubation with the purified enzyme. The effects of ammonium sulfate and organic solvents on the action pattern of the thermostable chitosanase were investigated. The amounts of (GlcN)3-(GlcN)6 were increased about 30% (w/w) in DAC 99 soluble chitosan containing 10% ammonium sulfate, and (GlcN)1 was not produced. The monophasic reaction system consisted of DAC 72 soluble chitosan in 10% EtOH also showed no formation of (GlcN)1, however, the yield of (GlcN)3 approximately (GlcN)6 was lower than DAC 99 soluble chitosan-10% ammonium sulfate. The optimal concentration of ammonium sulfate to be added was 20%. At this concentration, the amount of hexamer was increased by over 12% compared to the water-salt free system. PMID- 11388457 TI - Identification of 2,3-dihydro-gamma-ionylideneethanol in Cercospora cruenta. AB - During our scrutiny of GC-EI-MS date for C15 alcohols as putative intermediates on the ABA biosynthetic pathway in Cercospora cruenta, a trace amount of 5-[2',2' dimethyl-6'-methylene-1'-cyclohexyl]-3-methyl-4-penten-1-ol (2,3-dihydro-gamma ionylideneethanol) was identified. Feeding experiments indicated that this compound was not an intermediate to ABA, but a catabolite that originated from gamma-ionylideneacetaldehyde. The stereochemistry of 2,3-dihydro-gamma ionylideneethanol was deduced to be (3R,1'S) from a comparison with an authentic specimen prepared via baker's yeast asymmetric reduction. PMID- 11388458 TI - Biological evaluation of 5-substituted pyrimidine derivatives as inhibitors of brassinosteroid biosynthesis. AB - A series of 5-substituted pyrimidine derivatives was synthesized, and their ability to inhibit brassinosteroid biosynthesis was tested. The biological activity of these compounds was evaluated by the cress stem elongation method. Among the synthesized compounds, alpha-(4-chlorophenyl)-alpha-phenyl-5 pyrimidinemethanol (DPPM 4) exhibited potent inhibitory activity for retarding cress stem elongation in the light. This inhibition was reversed by the application of 10 nM brassinolide, but not by 1 microM GA3. DPPM 4 also affected Arabidopsis growth in the dark. DPPM 4-treated Arabidopsis had phenotypes like those of brassinosteroid-deficient mutants, with short hypocotyls and open cotyledons, in the dark. These biological changes were restored by the co application of 10 nM brassinolide, but not by 1 microM GA3, suggesting that the primary site of action of DPPM 4 was the brassinosteroid biosynthetic pathway. PMID- 11388459 TI - Incorporation of the whole chromosomal DNA in protoplast lysates into competent cells of Bacillus subtilis. AB - Competent cells of Bacillus subtilis AC870 (purB, leuB, trpC, ald-1) were transformed to Ade+, Trp+, or Ade+ Trp+ with DNA in protoplast lysates of B. subtilis AC819 (hisH, tet-1, rpsL, smo-1). The cotransfer ratio of purB to trpC was constant at 7-9% (Ade+ Trp+/Trp+) or 3% (Ade+ Trp+/Ade+) at protoplast concentrations of 2.7 x 10(3) to approximately 2.7 x 10(6) per ml. The whole chromosomal DNA must be certainly incorporated into competent cells from the following reasons; (1) purB is opposite to trpC on the chromosome, (2) 2.7 x 10(3) protoplasts per ml is about 100 times lower than 3.2 x 10(5) competent cells per ml, and (3) the cotransfer ratio is constant at all the concentrations. Similar results were obtained with the cotransfer ratio of purA to trpC. The transformation requires several Com proteins including ComK. PMID- 11388460 TI - Cloning, sequence analysis, and expression in Escherichia coli of the gene encoding monovalent cation-activated levodione reductase from Corynebacterium aquaticum M-13. AB - The gene encoding (6R)-2,2,6-trimethyl-1,4-cyclohexanedione (levodione) reductase was cloned from the genomic DNA of the soil isolate bacterium Corynebacterium aquaticum M-13. The gene contained an open reading frame consisting of 801 nucleotides corresponding to 267 amino acid residues. The deduced amino acid sequence showed approximately 35% identity with other short chain alcohol dehydrogenase/reductase (SDR) superfamily enzymes. The probable NADH-binding site and three catalytic residues (Ser-Tyr-Lys) were conserved. The enzyme was sufficiently produced in recombinant Escherichia coli cells using an expression vector pKK223-3, and purified to homogeneity by two-column chromatography steps. The enzyme purified from E. coli catalyzed stereo- and regio-selective reduction of levodione, and was strongly activated by monovalent cations, such as K+, Na+, and NH4+, as was the case of that from C. aquaticum M-13. To our knowledge, this is the first sequencing report of a monovalent cation-activated SDR enzyme. PMID- 11388461 TI - Purification of soluble beta-glucan with immune-enhancing activity from the cell wall of yeast. AB - Beta-glucan, one of the major cell wall components of Saccharomyces cerevisiae, has been found to enhance immune functions, especially by activating macrophages. However, a major obstacle to the clinical application of beta-(1-->3)-glucan is its low solubility in aqueous media. In this study, soluble beta-glucan, free of mannoprotein, was prepared, and its effects on TNF-alpha secretion and phagocytosis by macrophages were evaluated. Beta-glucan was first rendered soluble from the yeast cell wall by alkaline extraction (glucan-p1). The extract contained 2.8% of protein which was subsequently removed by successive DEAE cellulose and ConA chromatography. Beta-glucan thus prepared was completely free of mannoprotein and was soluble at neutral pH (glucan-p3). The effects of beta glucan on phagocytosis and TNF-alpha release activity were investigated. While glucan-p1 moderately induced TNF-alpha secretion at 200 microg/ml (550 pg of TNF alpha/5 x 10(5) cells), glucan-p3 markedly stimulated macrophages at 200 microg/ml (2,860 pg of TNF-alpha/5 x 10(5) cells). Furthermore, glucan-p3 stimulated phagocytosis about 20% more than glucan-p1 did. In conclusion, we purified water-soluble beta-glucan which was completely devoid of mannoprotein and effectively stimulated the macrophage function, enabling it to be used as an intravenous injection for sepsis. PMID- 11388462 TI - Purification and properties of a galacturonic acid-releasing exopolygalacturonase from a strain of Bacillus. AB - An exopolygalacturonase [exo-PGase; poly (1,4-alpha-D-galacturonide) galacturonohydrolase, EC 3.2.1.67] was found to be extracellularly produced by Bacillus sp. strain KSM-P443. The exo-PGase was purified to homogeneity, as judged by polyacrylamide gel electrophoresis, through sequential column chromatographies. The enzyme had a molecular weight of approximately 45,000 and an isoelectric point of pH 5.8. The N-terminal sequence was Ser-Met-Gln-Lys-Ile Lys-Asp-Glu-Ile-Leu-Lys-Thr-Leu-Lys-Val-Pro-Val-Phe and had no sequence similarity to those of other pectinolytic enzymes reported to date. Maximum activity toward polygalacturonic acid (PGA) was observed at 60 degrees C and at pH 7.0 in 100 mM Tris-HCl buffer without requiring any metal ions. When the chain length of oligogalacturonic acids increased, the apparent Km for them decreased, but the kcat values increased. This is the first bacterial exo-PGase that releases exclusively mono-galacturonic acid from PGA, di-, tri-, tetra-, and penta-galacturonic acids. PMID- 11388463 TI - Non-involvement of the K-ras mutation in colon carcinogenesis promoted by dietary deoxycholate in azoxymethane-treated rats. AB - Fisher-344 rats, whose ileum or jejunum had been surgically removed to change the influx of bile acids into the colon, were intraperitoneally administered with azoxymethane and fed on a diet containing deoxycholate for 39 weeks to induce colon cancer. Fecal bile acids in the ileum-resected group were 1.5-times and serum bile acids were about half of those in the jejunum-resected group. As a result, the incidence and number of tumors were higher in the ileum-resected group. In the total of 59 colon tumors (40 were in the ileum-resected group and 19 in the jejunum-resected group), 56 were carcinomas, including two well differentiated invasive and two mucinous carcinomas found in the ileum-resected rats. However, only three carcinomas, two invasive and one non-invasive, had the K-ras mutation. These results demonstrate that the K-ras mutation was not essentially involved in deoxycholate-promoted colon carcinogenesis. PMID- 11388464 TI - Increases of secondary metabolite production in various plant cell cultures by co cultivation with cork. AB - Cork tissues increased secondary metabolite production of various plant cell cultures in a different manner from those of conventional elicitors. In Sophora flavescens and Glycyrrhiza glabra cultured cells, cork tissues increased the amounts of both lipophilic and hydrophilic flavonoids without affecting the cell growth, although elicitors such as copper ion and yeast extracts showed a clear inhibition of cell growth with the increasing amount of these lipophilic ones. The validity of this effect of cork tissues covered a wide range of aromatic compounds produced by suspension cell cultures derived from diverse plant species. Woody tissues of Japanese cypress had a very similar effect to that of cork. Partial purification of cork tissues suggested that the production stimulating factor was present in the hemicellulose B fraction that was not included in the dedifferentiated cultured tissues. PMID- 11388465 TI - Biosynthesis of fattiviracin FV-8, an antiviral agent. AB - Streptomyces microflavus strain No. 2445 produces many derivatives of fattiviracin antibiotics. The major product of these derivatives is fattiviracin FV-8, which consists of four glucose and two trihydroxy fatty acid residues. We found that this strain has the ability to convert several sugars in the culture medium to glucose, and the glucose added to the medium is directly incorporated into the FV-8 molecule. Two trihydroxy fatty acid residues in the FV-8 molecule are derived from acetic acid, and production of FV-8 is inhibited by the addition of cerulenin, which is an inhibitor of fatty acid biosynthesis. PMID- 11388466 TI - Increase in the stability of serine acetyltransferase from Escherichia coli against cold inactivation and proteolysis by forming a bienzyme complex. AB - Cysteine synthetase from Escherichia coli is a bienzyme complex composed of serine acetyltransferase (SAT) and O-acetylserine sulfhydrylase-A (OASS). The effects of the complex formation on the stability of SAT against cold inactivation and proteolysis were investigated. SAT was reversibly inactivated on cooling to 0 degrees C. Ultracentrifugal analysis showed that SAT (a hexamer) was dissociated mostly into two trimers on cooling to 0 degrees C in the absence of OASS, while in the presence of OASS one trimer of the SAT subunits formed a complex with one dimer of OASS subunits. In the presence of OASS, not only the cold inactivation rate was reduced but also the reactivation rate was increased. Furthermore, SAT became stable against proteolytic attack by alpha-chymotrypsin and V8 protease by forming the complex with OASS. On the other hand, SAT was degraded by trypsin in the same manner both in the presence and in the absence of OASS. The different tendency in the stability against proteolysis with the different proteases was discussed with respect to the substrate specificity of the proteases and amino acid sequence of the C-terminal region of SAT that interacts with OASS. PMID- 11388468 TI - Novel method using phytase for separating soybean beta-conglycinin and glycinin. AB - A novel method for separating soybean beta-conglycinin and glycinin from defatted soymilk by a phytase treatment was developed. Phytase was added to defatted soymilk (1000FYT/100 g of protein) at pH 6.0, and the mixture incubated for 1 h at 40 degrees C. This procedure separated beta-conglycinin and glycinin without needing a reducing agent or cooling into the soluble and insoluble fractions, respectively. Simultaneously, most of the phytate in both proteins was removed. PMID- 11388467 TI - Transglycosylation to ginseng saponins by cyclomaltodextrin glucanotransferases. AB - Ten new alpha-glucosylginsenosides were found to be synthesized from dextrin and four ginsenosides, -Rb1, -Rc, -Re, and -Rg1, by the successive actions of B. stearothermophilus cyclomaltodextrin glucanotransferase and Rhizopus glucoamylase. Seven of them were isolated in the pure state by extraction with n butanol saturated with water, silica gel column chromatography, and high pressure liquid chromatography, and identified as 3-O-[alpha-D-glcp-(1-->4)-beta-D-glcp-(1 ->2)-beta-D-glcp]-20-O-[beta-D-glcp-(1-->6)-beta-D-glcp]-20(S)-protopanaxadiol, 3 O-[beta-D-glcp-(1-->2)-beta-D-glcp]-20-O-[alpha-D-glcp-(1-->4)-beta-D-glcp-(1- >6)-beta-D-glcp]-20(S)-protopanaxadiol, 3-O-[alpha-D-glcp-(1-->4)-beta-D-glcp-(1- >2)-beta-D-glcp]-20-O-[alpha-L-araf-(1-->6)-beta-D-glcp]-20(S)-protopanaxadiol, 3 O-[beta-D-glcp-(1-->2)-beta-D-glcp]-20-O-[(4G-alpha-D-glcp)-alpha-L-araf-(1-->6) beta-D-glcp]-20(S)-protopanaxadiol, 6-O-[alpha-L-rhap-(1-->2)-beta-D-glcp]-20-O [alpha-D-glcp-(1-->4)-beta-D-glcp]-20(S)-protopanaxatriol, 6-O-[alpha-D-glcp-(1- >4)-beta-D-glcp]-20-O-(beta-D-glcp)-20(S)-protopanaxatriol, and 6-O-[alpha-D-glcp (1-->3)-beta-D-glcp]-20-O-(beta-D-glcp)-20(S)-protopanaxatriol, by spectroscopy (FAB-MS, IR, 1H-NMR and 13C-NMR) and hydrolysis products in 50% acetic acid. The bitterness of these alpha-glucosyl-ginsenosides was less than that of ginsenosides. PMID- 11388469 TI - A novel cryoprotective protein (CRP) with high activity from the ice-nucleating bacterium, Pantoea agglomerans IFO12686. AB - The ice-nucleating bacterium, Pantoea agglomerans IFO12686, induces the cryoprotective protein (CRP) by cold acclimation at 12 degrees C. The CRP was purified to apparent homogeneity by various chromatographies. We found that the purified CRP was a monomer of approximately 29,000 according to gel filtration chromatography and SDS-PAGE, and was a heat-stable protein. The CRP could protect freeze-labile enzymes, lactate dehydrogenase (LDH), alcohol dehydrogenase (ADH) and isocitrate dehydrogenase (iCDH), against freezing-thawing denaturation. The activity of the CRP was about 3.5 x 10(4) times more effective than bovine serum albumin (BSA) and 2 x 10(6) times than COR26 from the ice-nucleating bacterium Pseudomonas fluorescens KUIN-1. We confirmed that the CRP was a novel protein, as judged by the a different molecule mass from the already-known cryoprotectants, and has an extremely high cryoprotective activity. PMID- 11388470 TI - Characterization of soluble protein extracts from keratinized tissues: identification of ubiquitin universally distributed in hair, nail, and stratum corneum. AB - Partial protein extracts were prepared from hair, nail, and stratum corneum in the absence of urea and interfacial surfactant. Tricine-sodium dodecyl sulfate polyacrylamide gel electrophoreses of these extracts showed low-molecular weight protein-rich patterns apparently different from those of whole protein extracts, which mainly consist of keratin bands. Several protein bands characterized each keratinized tissue or its derived species. In addition, we identified a major band of approximately 7 kDa as ubiquitin, a ubiquitously distributed protein that mediates non-lysosomal protein degradation, through direct amino acid sequence analysis of the electro-blotted protein band. The partial extraction is useful for investigation of soluble proteins retained in the keratinized tissues. PMID- 11388471 TI - Structures of the N-linked sugar chains in PAS-7 glycoprotein sharing the same protein core with PAS-6 glycoprotein from the bovine milk fat globule membrane. AB - Glycoproteins PAS-6 (50 kDa) and -7 (47 kDa) from the bovine milk fat globule membrane share a common protein core but differ in their carbohydrate moiety. We here analyzed and proposed the structures of the N-linked sugar chains of PAS-7. The N-linked sugar chains were liberated from PAS-7 by hydrazinolysis and, after modifying the reducing ends with 2-aminopyridine (PA), were separated into one neutral (7N, 55%) and two acidic (7M, mono-, 43%; 7D, di-, 2%) sugar chain groups. The latter were converted into neutral groups (7MN and 7DN) by sialidase digestion. 7N was finally separated into 5 chains (7N1A, 7N1B-1, 7N1B-2, 7N2A, and 7N2B), and 7MN and 7DN were separated into 3 (7MN1, 7MN2, and 7MN3) and 2 (7DN1 and 7DN2) chains, respectively. The structure of each of these PA-neutral sugar chains was determined by sugar analysis, sequential exoglycosidase digestion, partial acetolysis, and 1H-NMR spectroscopy. The results show that the 10 sugar chains were of the biantennary complex type with and without fucose. The structure of 7N2A, one of the major sugar chains, was proposed as; [structure: see text] A structural comparison between PAS-6 and -7 indicated that, although they shared the same protein core, their sugar moiety was markedly different, involving the existence of a different pathway during the post-transcriptional modification. PMID- 11388472 TI - Controlled trial of the effects of milk basic protein (MBP) supplementation on bone metabolism in healthy adult women. AB - Milk has more beneficial effects on bone health compared to other food sources. Recent in vitro and in vivo studies showed that milk whey protein, especially its basic protein fraction, contains several components capable of both promoting bone formation and inhibiting bone resorption. However, the effects of milk basic protein (MBP) on bone metabolism of humans are not known. The object of this study was to examine the effects of MBP on bone metabolism of healthy adult women. Thirty-three normal healthy women were randomly assigned to treatment with either placebo or MBP (40 mg per day) for six months. The bone mineral density (BMD) of the left calcaneus of each subject was measured at the beginning of the study and after six months of treatment, by dual-energy x-ray absorptiometry. Serum and urine indices of bone metabolism were measured at the base line, three month intervals, and the end of the study. Daily intake of nutrients was monitored by a three-day food record made at three and six months. The mean (+/- SD) rate of left calcaneus BMD gain of women in the MBP group (3.42 +/- 2.05%) was significantly higher than that of women in the placebo group (2.01 +/- 1.75%, P = 0.042). As compared with the placebo group, urinary cross-linked N teleopeptides of type-I collagen/creatinine and deoxypyridinoline/creatinine were significantly decreased in the MBP group (p < 0.05), while no significant differences between the two groups were observed in serum osteocalcin and bone specific alkaline phosphatase concentrations. A daily MBP supplementation of 40 mg in healthy adult women can significantly increase their BMD independent of dietary intake of minerals and vitamins. This increase in BMD might be primarily mediated through inhibition of osteoclast-mediated bone resorption by the MBP supplementation. PMID- 11388473 TI - Structural elucidation of twelve novel esters composed of five fatty acids and three new branched alcohols together with four monoterpenoids from Sancassania shanghaiensis (Acari: Acaridae). AB - A total of 12 novel esters and four monoterpenoids (rosefuran, (2R,3R) epoxyneral, and alpha- and beta-acaridials) were detected by GC/MS analyses as the opisthonotal gland components of Sancassania shanghaiensis. The acidic fraction after hydrolysis was composed of five common fatty acids (palmitic, stearic, oleic, linoleic and arachidic acid), while the alcoholic fraction consisted of two major components (C6 and C8 alcohols with branched methyls), together with a trace amount of C9 alcohol. The two major alcohols were identified as new alcohols [(S)-2-methylpentanol and (2S,4S)-2,4-dimethylhexanol] by comparing the physico-chemical data of their 3,5-dinitrobenzoates with those of regio-selectively synthesized alcohols. The C9 alcohol was suggested as (2S,4S)-2,4-dimethylheptanol, based on a structural and biogenetic analogy to the C6 and C8 alcohols. Five of the compounds were each identified by GC to be (S)-2 methylpentyl esters from five fatty acids, and the other five components likewise as (2S,4S)-2,4-dimethylhexyl esters. The remaining two were suggested as (2S,4S) 2,4-dimethylheptyl stearate and linolate. PMID- 11388474 TI - Greater effect of dietary potassium tripolyphosphate than of potassium dihydrogenphosphate on the nephrocalcinosis and proximal tubular function in female rats from the intake of a high-phosphorus diet. AB - We examined whether a difference in potassium dihydrogenphosphate (KH2PO4) and potassium tripolyphosphate (K5P3O10) as dietary phosphorus sources could differentially effect the nephrocalcinosis and proximal tubular function in female rats. Rats were fed on a diet containing KH2PO4 or K5P3O10, at the normal phosphorus level (normal phosphorus diet) or at a high phosphorus level (high phosphorus diet) for 21 d. Nephrocalcinosis, as confirmed by a histological examination, was apparent in all rats fed on the high-phosphorus diet, and this condition was more severe in those rats fed on K5P3O10 than in those fed on KH2PO4. As indicators of the proximal tubular function, the N-acetyl-beta-D glucosaminidase activity in urine and the urinary beta2-microglobulin excretion were significantly increased in those rats fed on the high-phosphorus diet containing K5P3O10. These results indicate that the intake of a high-phosphorus diet, more strongly influenced the nephrocalcinosis and proximal tubular function when K5P3O10 rather than KH2PO4 was used as the dietary phosphorus source. PMID- 11388475 TI - Different response to choline deficiency of the serum ornithine carbamoyltransferase activity in four strains of rats. AB - Rats of the Donryu, Wistar, Fischer, and Sprague-Dawley strains were examined for the effects of choline deficiency on liver lipids, serum lipids, and serum ornithine carbamoyltransferase. The liver total lipid, triacylglycerol, cholesterol and phospholipid contents in the choline-deficient rats were significantly higher than those in choline-sufficient rats. The contents of total lipids and phospholipids in the liver of the Wistar and Fischer rats fed on a choline-deficient diet were significantly higher than those of the Donryu and Sprague-Dawley rats. The levels of triacylglycerol, cholesterol and phospholipids in the serum were significantly decreased by feeding with the choline-deficient diet. The serum ornithine carbamoyltransferase activity was increased in the Wistar and Fischer strains by feeding with the choline-deficient diet. The Wistar and Fischer strains were consequently the most sensitive to both lipid accumulation and liver lesions induced by the choline deficiency. PMID- 11388476 TI - Detection of antifungal activity in Belamcanda chinensis by a single-cell bioassay method and isolation of its active compound, tectorigenin. AB - The antifungal activity of Belamcanda chinensis was evaluated by a single-cell bioassay method. An active fraction was separated by silica gel column chromatography and reverse-phase HPLC. The isolated compound was found to be identical to tectorigenin (5,7-dihydroxy-3-(4-hydroxy phenyl)-6-methoxy-4H-1 benzopyran-4-one) which has formerly appeared in the literature without any remarks on its antimicrobial activity. Antimicrobial activity was investigated against 17 strains of fungi and 6 strains of bacteria. This compound showed marked antifungal activity against dermatophytes of the genera Trichophyton, the minimum inhibitory concentration (MIC) being in the range of 3.12-6.25 mg/ml. PMID- 11388477 TI - Microbial hydroxylation of (+/-)- and (-)-(2Z,4E)-5-(1',2'-epoxy-2',6',6' trimethylcyclohexyl)-3-methyl-2,4-pentadienoic acid into (+/-)- and (-)-xanthoxin acid by Cunninghamella echinulata. AB - Microbial hydroxylation of (+/-)-(2Z,4E)-5-(1',2'-epoxy-2',6',6' trimethylcyclohexyl)-3-methyl-2,4-pentadienoic acid (3a) with Cercospora cruenta, a fungus producing (+)-abscisic acid, gave a four-stereoisomeric mixture consisting of (+)- and (-)-xanthoxin acid (4a), and (+)- and (-)-epi-xanthoxin acid (5a) by an HPLC analysis with a chiral column. Screening of the microorganisms capable of oxidizing (+/-)-3a showed that Cunninghamella echinulata stereoselectively oxidized (+/-)-3a to xanthoxin acid (4a) with the some degree of enantioselectivity as (-)-3a to (-)-4a. PMID- 11388478 TI - Anti-tumor promoting activity of bufadienolides from Kalanchoe pinnata and K. daigremontiana x tubiflora. AB - Five bufadienolides (1-5) isolated from the leaves of Kalanchoe pinnata and K. daigremontiana x tubiflora (Crassulaceae) were examined for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation in Raji cells induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate. All bufadienolides showed inhibitory activity, and bryophyllin A (1) exhibited the most marked inhibition (IC50 = 0.4 microM) among the tested compounds. Bryophyllin C (2), a reduction analogue of 1, and bersaldegenin-3-acetate (3) lacking the orthoacetate moiety were less active. These results strongly suggest that bufadienolides are potential cancer chemopreventive agents. PMID- 11388479 TI - Isolation and identification of trans-2- and trans-3-hydroxy-1,8-cineole glucosides from Alpinia galanga. AB - Three hydroxy-1,8-cineole glucopyranosides, (1R, 2R, 4S)- and (1S, 2S, 4R)-trans 2-hydroxy-1,8-cineole beta-D-glucopyranosides, and (1R, 3S, 4S)-trans-3-hydroxy 1,8-cineole beta-D-glucopyranoside, which are possible precursors of acetoxy-1,8 cineoles as unique aroma components, were isolated from the rhizomes of greater galangal (Alpinia galanga W.). Their structures were analyzed by FAB-MS and NMR spectrometry, and the absolute configulation of each aglycone was determined by using a GC-MS analysis with a capillary column coated with a chiral stationary phase. The composition of the diastereomers of (1R, 2R, 4S)- and (1S, 2S, 4R) trans-2-hydroxy-1,8-cineole beta-D-glucopyranosides in the rhizomes was determined as 3:7 by a GC-MS analysis after preparing the trifluoroacetate derivatives of the glucosides. PMID- 11388480 TI - Wine has activity against entero-pathogenic bacteria in vitro but not in vivo. AB - We studied the activity of wine against entero-pathogenic bacteria both in vitro and in vivo. The food-borne bacteria were killed in both red and white wine within 30 min. However, the results of a Salmonella infection experiment using mice suggested that wine was not effective in preventing food-borne diseases in vivo. PMID- 11388481 TI - Rapid dephosphorylation of eIF4E by dietary protein in the skeletal muscle and liver of food-deprived rats. AB - The effect of dietary protein on eIF4E phosphorylation was examined in rats starved for 18 h and then fed on either a 20% casein diet (20C) or a protein-free diet (0C). Refeeding with the 20C diet, but not the 0C diet, resulted in partial dephosphorylation of eIF4E in both the skeletal muscle and liver. The results suggest that the dephosphorylation of eIF4E in response to food intake was regulated by the increase in plasma amino acid concentration that occurred after feeding with the 20C diet. PMID- 11388482 TI - Effect of zinc deficiency on betacyanin production in a cell suspension culture of table beet (Beta vulgaris L.). AB - The effect of microelements in the Linsmaier-Skoog (LS) medium on betacyanin production was investigated in suspension cultures of table beet (Beta vulgaris L.). Removing zinc from the medium resulted in a high betacyanin content of the cells, the betacyanin content of the cells decreasing with increasing zinc concentration in the medium. The betacyanin content of cells cultured in the medium without zinc was twice as high as that in the medium containing 0.03 mM of zinc. In the revised LS medium without zinc, the maximum betacyanin yield was obtained of 590 mg/l from a 21-day culture. PMID- 11388483 TI - Antibacterial activity of S-methyl methanethiosulfinate and S-methyl 2-propene-1 thiosulfinate from Chinese chive toward Escherichia coli O157:H7. AB - S-Methyl methanethiosufinate (1) and S-methyl 2-propene-1-thiosulfinate (2) were easily seperated from Chinese chive (Allium tuberosum L.) using simple column chromatography. Both compounds showed significant antibacterial activities against E. coli O-157:H7 including spoilage microorganism in food. Structural assignment was based on Mass and NMR-spectroscopic methods. PMID- 11388484 TI - Molecular cloning and functional expression of cDNA encoding the cysteine proteinase inhibitor Sca from sunflower seeds. AB - Sunflower cystatin a (Sca) is distinguished from other phytocystatins by its lack of the N-terminal about 20 amino acids, resulting in the absence of the evolutionarily conserved Gly residue. The cDNA encoding Sca was amplified by PCR methods. The cDNA consists of 520 nucleotides and includes an open reading frame encoding a polypeptide of 98 amino acids. Comparison of the deduced amino acid sequence with the Sca protein sequence indicated that the deduced sequence has an extra 15 amino acids and one amino acid at the N- and C-termini, respectively. This result suggests that Sca is synthesized as a preprotein (preSca) and proteolytic cleavages at peptide bonds may give rise to the mature Sca. To address this assumption and also to investigate the significance of the N terminal extension sequence to Sca for inhibitory activity, a recombinant pre-Sca (rpre-Sca), in which the N-terminal extension was fused to the matured Sca, and a recombinant matured Sca (rSca) were overproduced in Escherichia coli cells. Incubation of the rpre-Sca with a seed extract resulted in a mobility by SDS-PAGE that was the same as rSca, demonstrating a proteolytic cleavage by endogenous proteinases. The rSca and rpre-Sca proteins were further characterized with respect to inhibitory activity and sensorgrams of the interaction with papain. The result showed that rpre-Sca had stronger inhibitory activity than rSca, and that the increased activity toward papain was due to a lower dissociation rate constant. This finding indicates that the N-terminal region of rpre-Sca increases the inhibitory activity by stabilizing the rpre-Sca and papain complex. PMID- 11388485 TI - Facile synthesis of (R)-4-mercaptopyrrolidine-2-thione from L-aspartic acid. AB - An SN2-type of substitution of (S)-bromide 4, which had been prepared from L aspartic acid, with potassium thiobenzoate provided (R)-benzoylthio derivative 5 with complete inversion of the configuration. Compound 5 was converted, via iodide 6c, to (R)-4-amino-3-benzoylthiobutyric acid 8b. (R)-4-Mercapto pyrrolidine-2-thione 1 was readily obtained from 8b through cyclization with acetic anhydride, thionation with Lawesson's reagent and facile removal of the S benzoyl group with sodium methoxide. PMID- 11388486 TI - Cloning and characterization of a chitosanase gene from the koji mold Aspergillus oryzae strain IAM 2660. AB - A genomic copy of the gene coding for chitosanase (csnA) was isolated from Aspergillus oryzae IAM 2660. A. oryzae csnA contains an open reading frame that encodes a polypeptide of 245 amino acids with a calculated molecular mass of 26,500 Da. The deduced amino acid sequence of A. oryzae csnA indicates extensive similarities to those of other fungal chitosanases. PMID- 11388487 TI - Improvement of shark type I collagen with microbial transglutaminase in urea. AB - In the presence of urea, type I collagen could form a gel with crosslinks with microbial transglutaminase (MTGase). Collagen self-assembly was accelerated with the addition of MTGase. The proportion of reconstructed collagen fibrils was raised with the addition of MTGase. MTGase-treated collagen gel remained gelled at high temperatures at which collagen denatured. By treatment with MTGase, collagen could form the gel under impossible condition to collagen self-assembly, and that denaturation temperature was raised. PMID- 11388488 TI - Human milk bile-salt-stimulated lipase is extremely reactive with the monoclonal antibody 1CF11 which recognizes a human-specific carbohydrate antigen. AB - 1CF11 (Kanamaru, Y. et al.; Biochem. Biophys. Res. Commun., 249, 618-623, 1998) is a monoclonal antibody obtained after being raised in a mouse by injection of human milk MUC1 mucin as the antigen. Its reactivity was found to be unique in that it only reacts with a carbohydrate epitope shared by glycoproteins in human secretions, while its chemical nature is still unknown. Since a glycoprotein of Mr 135,000 (135K) in human milk was found to react extremely strongly with this antibody, we intended in this study to isolate the glycoprotein by a combination of various chromatographic techniques and identify it. It is a human milk bile salt-stimulated lipase. By comparison of its immunoreactivity and glycan structures so far reported with those of lactoferrin from human milk, it is suggested that the epitope recognized by mAb ICF11 could be a human-specific novel glycan. PMID- 11388489 TI - 7-Hydroxyphthalide: a new natural salicylaldehyde analog from Oulenzia sp. (Astigmata: Winterschmitiidae). AB - A new salicyl lactone was detected from the unidentified Oulenzia sp. and its chemical structure was elucidated as 7-hydroxyphthalide (7-hydroxy-3H isobenzofuran-1-one), based on its GC/MS and GC/FT-IR data and SiO2 column behavior. The compound was synthesized by NaBH4 reduction of 3-hydroxyphthalic anhydride. The GC/MS and GC/FT-IR spectra of the natural compound were consistent with those of the synthetic product. Although the compound is known as a medical material, this is the first example of its presence in nature. PMID- 11388490 TI - Expression of marY1, a gypsy-type LTR-retroelement from the ectomycorrhizal homobasidiomycete Tricholoma matsutake, in the budding yeast Saccharomyces cerevisiae. AB - marY1 is a gypsy-type LTR-retroelement present in the genome of the ectomycorrhizal homobasidiomycete Tricholoma matsutake. We document here that a marY1-lacZ gene fusion was expressed in the budding yeast Saccharomyces cerevisiae. The finding strongly suggests that marY1 is activated by trans regulatory factors common to higher fungi, and may be useful for the development of new recombinant systems in ectomycorrhizal fungi and homobasidiomycetes. PMID- 11388491 TI - Co-existing proteins interfere with the action of nordihydroguaiaretic acid on retrograde Golgi-to-ER protein trafficking in NRK cells and alpha-glucosidase reaction in vitro. AB - Induction of retrograde trafficking of mannosidase II and TGN38 in NRK cells and inhibition of alpha-glucosidase in vitro by nordihydroguaiaretic acid (NDGA) were strongly interfered with by serum, serum albumin, or other unrelated proteins added to the medium or incubation mixture. These observations indicate that NDGA interacts with diverse kinds of proteins, and therefore, pharmacological effects of NDGA at cellular levels should be carefully interpreted. PMID- 11388493 TI - Lifestyle and sociodemographic risk factors for death among middle-aged and elderly residents in japan from a five-year follow-up cohort study. AB - To examine the relationship between lifestyle and sociodemographic risk factors and mortality, a population-based prospective cohort study was conducted in two areas of Gunma Prefecture, Japan, and a cohort consisting of 11,565 subjects aged 40-69 at baseline in 1993 was followed. During the five-year follow-up period, 201 men and 103 women died. The relative risks (RRs) of risk factors were estimated by the Cox proportional-hazards model. Significant RRs with multivariate adjustment for all-cause mortality was observed for body mass index (BMI). The curve for the relationship between BMI and all-cause mortality was L shaped in men and U-shaped in women, with the lowest RRs at a BMI of 22-25 in both men and women. Other significant RRs for all-cause mortality were observed for obesity in the subjects' 30's in both men and women (RR: 2.42 and RR: 2.75), poor perceived health status in men (RR: 4.55), and having had a health examination in the past three years in both men and women (RR: 0.49 and RR: 0.46). These results suggested that increased risk of death was independently associated with a lower BMI, obesity in the subjects' 30's, and not undergoing health examinations, among both men and women, and poor perceived health status among men. PMID- 11388492 TI - Soybean resistant proteins interrupt an enterohepatic circulation of bile acids and suppress liver tumorigenesis induced by azoxymethane and dietary deoxycholate in rats. AB - We found that azoxymethane and dietary deoxycholate induced liver tumors in rats. The incidence and the development of the tumor were closely related to the enterohepatic circulation of bile acids. The feeding of a high-molecular-weight fraction of soy protein digest (HMF) suppressed the tumorigenesis, probably due to the inhibitory effect of soybean resistant protein on reabsorption of bile acids in the intestine. PMID- 11388494 TI - Magnitude of the regression to the mean within one-year intra-individual changes in serum lipid levels among Japanese male workers. AB - To investigate the magnitude of the 'regression to the mean' effect for the changes in serum total cholesterol (TC), triglycerides (TG), and high density lipoprotein cholesterol (HDLC) levels during one-year interval between annual health check-ups in occupational settings, the relationships between the baseline level and subsequent one-year change in TC, TG or HDLC were analyzed using paired health check-up data in 1998 and 1999 of 547 Japanese male clerical workers. After adjustment for age, body mass index (BMI), yearly change in BMI, drinking score and smoking score by the multivariate analyses, the yearly changes in each serum lipid (deltaTC, deltaLn(TG) or deltaHDLC) were clearly inversely associated with the lipid levels in 1998. For example, in the multiple linear regression analyses setting delta value in each serum lipid as a dependent variable, the partial regression coefficients for the baseline lipid levels (beta1) were - 0.21 (p<0.001) for the TC, -0.39 (p<0.001) for the Ln(TG) and -0.15 (p<0.001) for the HDLC, respectively. These results suggest that the observed yearly change in each serum lipid level may largely reflect the 'regression to the mean' effect in addition to the real yearly biological change. PMID- 11388495 TI - Response bias by neuroblastoma screening participation status and social desirability bias in an anonymous postal survey, Ishikawa, Japan. AB - OBJECTIVE: To examine response bias by neuroblastoma screening participation status in a population-based postal survey of parents in Ishikawa Prefecture, Japan. METHODS: The eligibility criteria for the study were: 1) parents whose infants were born in Ishikawa Prefecture between March 1997 and February 1998, and 2) of those parents who resided in the Prefecture in March 1999. Four-page questionnaires were mailed to one-third of screening participants (n = 2,886) and all the nonparticipants (n = 1,401). Questionnaires were anonymous, with no identifiers on the questionnaire. Colored papers were used for printing questionnaires to differentiate screening participation status. Response rates were calculated using demographic information on the infant registry as the denominator and demographic characteristics data from the returned questionnaire as the numerator. RESULTS: The response rate was 63% for participants and 33% for nonparticipants. The following factors were associated with lower response rates regardless of screening participation status: older maternal age (> or = 35 years), higher parity (> or = 4), nuclear family status, and mother having a full time occupation. Approximately 20% of screening nonparticipants reported having participated in the screening. Place of residence, maternal age, and parity were associated with the percentage of incorrect reporting. CONCLUSION: Screening participation status was a major factor associated with low response rate, although some demographic characteristics were also predictive of low response rates. Incorrect reporting of screening participation among nonparticipants indicates a strong social desirability bias in this official survey in Japan. PMID- 11388496 TI - Occupational health and safety in the least developed countries--a simple case of neglect. AB - In many of the least developed countries, working people are significantly exposed to a number of occupational problems that may result in a deterioration of their health, safety and well being. These work-related problems are untenable, not only because of the occupational problems itself but also because of the simultaneous exposure to heat, dusts, noise, organo-chemicals, and biological and environmental pollution. This situation has existed for a long time due to various socio economic,geographical, cultural and local factors. The deteriorating situation of health and safety in the workplace may perhaps exist due to the inadequate resource facilities, economic constraints and lack of opportunity to conduct research and studies on the assessment of exposure diseases associations. Officials, who are employed by the state, are not able to implement work regulations and labour legislation easily. Generally, they are not professionally trained and expert in the occupational health, industrial hygiene and/or safety fields, and thus, successful application and implementation of control measures are lacking. Steps to control work exposure limits have been ineffective, since national policies have been rare, owing to the multiple obstacles in preventing occupational problems. However, the major focus is on practical solutions to differing workers' needs, consideration of which is very important, depending on the what the industrial entrepreneurs could reasonably to be expected to afford. Why there is a lack of motivation and effort regarding the development of health and safety-this paper explores some important issues, aiming to focus public attention on the legacy of national and international efforts. Examples are likewise given to show the real situation of health and safety in the least developed countries. PMID- 11388497 TI - Smoking, alcohol, sleep and risk of idiopathic sudden deafness: a case-control study using pooled controls. AB - Sudden deafness sometimes has an identifiable cause, but in most cases the cause is unknown (idiopathic sudden deafness). Vascular impairment has been proposed as an aetiological mechanism for this condition, but it is unclear whether traditional cardiovascular risk factors, such as smoking or alcohol intake, are associated with this condition. We accordingly investigated associations of idiopathic sudden deafness with smoking, alcohol intake and sleep duration in a case-control study. Cases were consecutive patients diagnosed with idiopathic sudden deafness between October 1996 and August 1998 at collaborating hospitals in Japan. Controls were obtained from a nationwide database of pooled controls, with matching for age, gender and residential district. Exposure variables were assessed from a self-administered questionnaire. Subgroup analyses were performed using audiometric subtypes of sudden deafness. Data were obtained for 164 cases and 20,313 controls. Increased risks of idiopathic sudden deafness were observed among participants who consumed two or more units of alcohol per day (OR=1.90, 95% CI=1.12-3.21), and among participants who slept less than seven hours per night (OR=1.61, 95% CI=1.09-2.37). The direct association with alcohol intake was particularly strong for the participants with profound hearing loss. There was little evidence of an association with smoking. This study suggests that alcohol intake and short sleep duration might be risk factors for idiopathic sudden deafness. PMID- 11388498 TI - Meta-analysis for epidemiologic studies on the relationship between smoking and Parkinson's disease. AB - Many epidemiologic studies on the relationship between smoking and Parkinson's disease (PD) have been conducted. Morens et al. reviewed many articles in the study field and concluded that smoking is inversely associated with the risk of PD. In the present study, the object is to obtain summarized risk estimates of the relationship from the published articles using meta-analysis. Summarized risk estimates on the relationship between smoking and PD were found to be about 0.5 with statistical significance in meta-analysis. Therefore, the result that smoking is inversely associated with the risk of PD is appropriate. PMID- 11388499 TI - The role of epidemiology in challenging the HIV/AIDS pandemic. AB - The HIV/AIDS pandemic has challenged the resourcefulness of epidemiology and epidemiologists. In response to the challenge, epidemiologists have used existing epidemiologic strategies, expanded existing strategies, and developed new strategies to answer key questions about the transmission of HIV, the natural history of HIV at the molecular, host, and community levels, for evaluation of treatment effectiveness and intervention strategies, and to inform public health policy. In responding to the challenge of the pandemic, epidemiologists have also increasingly collaborated with scientists from other disciplines, particularly immunology, virology, and the behavioral sciences. Examples of the application of these epidemiologic strategies are presented. PMID- 11388500 TI - Progressive chronic Chagas heart disease ten years after treatment with anti Trypanosoma cruzi nitroderivatives. AB - A randomized ten-year follow-up study involving 91 Chagas patients and 41 uninfected controls was undertaken to determine the effectiveness of nitroderivative therapy. Anti-Trypanosoma cruzi antibodies were consistently lower one year after treatment than 10 years thereafter (P < 0.001). The blood of all treated and 93.7% of untreated Chagas patients yielded polymerase chain reaction (PCR) product from probes annealing to T. cruzi nuclear DNA, indicating active infection. Competitive PCR showed means +/- standard deviations of 20.1+/ 22.6 T. cruzi/ml of blood from untreated and 13.8+/-14.9 from treated Chagas patients, but the differences between means were not statistically significant (P > 0.05). Electrocardiograms recorded a gamut of alterations several-fold more frequent in Chagas patients, regardless of treatment, than in uninfected controls (P < 0.001). These results show that nitroderivative therapy for T. cruzi infections is unsatisfactory and cannot be recommended since it fails to eradicate the parasite or change the progression of heart disease in chronic Chagas patients. PMID- 11388501 TI - Short report: failure to select for chloroquine- or mefloquine-resistant Plasmodium berghei through drug pressure in Anopheles stephensi mosquitoes. AB - We investigated whether chloroquine- or mefloquine-resistant Plasmodium berghei could be selected through drug pressure applied during continuous cyclical transmission in Anopheles stephensi mosquitoes. Mosquitoes were infected by feeding them on mice previously inoculated with a drug-sensitive clone of P. berghei ANKA. Mosquitoes ingested mefloquine or chloroquine with the infectious blood-meal, or by feeding on a drug-treated (uninfected) mouse 4 or 10 days after the infectious blood-meal. Twenty-two days after being infected, mosquitoes transmitted sporozoites to uninfected mice. Blood from these animals was used to infect naive mice that were then used to reinitiate the mouse/mosquito/mouse cycle. A total of 20 passages through mosquitoes were completed while under drug pressure. Drug-resistance levels were assessed in the initial clone and after 20 passages through mosquitoes. None of 18 "sub-clones" of parasites showed significant increases in chloroquine or mefloquine resistance, suggesting that exposure of sporogonic stage Plasmodium to chloroquine or mefloquine will not result in the development of drug resistance. PMID- 11388502 TI - Therapy for human gastrointestinal microsporidiosis. AB - Gastrointestinal microsporidiosis is a major cause of diarrhea and wasting in persons with acquired immune deficiency syndrome (AIDS). Microsporidia demonstrate properties of both true eukaryotes and prokaryotes. The biology of microsporidia makes its elimination from the gastrointestinal tract therapeutically challenging. This organism depends greatly on the host for its energy needs and reproduction; microsporidial spores are impervious to the elements. Microsporidial infection of the gastrointestinal tract, principally with Enterocytozoon bieneusi and Encephalitozoon intestinalis in patients with AIDS has been treated with different medical regimens with variable success. The less common pathogen, E. intestinalis, responds well to albendazole, making it excellent first-line therapy, but such is not the case for E. bieneusi. None of the benzimidazoles has been demonstrated to be efficacious for E. bieneusi. On the other hand, E. bieneusi has shown excellent clinical therapeutic response to either direct action with fumagillin or its analogue, TNP-470, or indirectly by immune enhancement by suppression of the HIV virus with more aggressive, highly effective antiretroviral therapy. Further work is necessary to fully establish proper therapeutic protocols and manage side effects of the treatments. Other promising forms of therapy such as polyamine inhibitors and thalidomide demonstrate certain effectiveness in treatment of microsporidian in vitro (polyamine inhibitors) and in selected cases in vivo (thalidomide). Lack of either sufficiently suggestive or definitive human studies prevents the endorsement of these modes of therapy for treatment of gastrointestinal microsporidiosis at this time. PMID- 11388503 TI - IgE deposition in brain microvessels and on parasitized erythrocytes from cerebral malaria patients. AB - Postmortem brain tissues of 21 cerebral malaria cases were obtained in Myanmar and Vietnam. The tissues were examined by light microscopy and by an immunohistochemical method. Brain microvessels (capillaries and venules) were examined for the presence of immunoglobulins IgE and IgG, Plasmodium falciparum antigen, and parasitized erythrocytes (PRBC). Deposition of IgE, IgG, and P. falciparum antigen was observed in the microvessels from all specimens examined. Sequestered PRBC in the microvessels were positive for IgG in all 21 cases and for IgE in six cases. In the latter cases, the percentage of microvessels with sequestered PRBC was > 50%, with the frequency of IgE-positive cells ranging from 42% to 52%. In contrast, in five cases that were only weakly positive for IgE, the percentage of microvessels with sequestered PRBC was remarkably low (< 1%). These data indicate that the degree of deposition of IgE in microvessels and on PRBC from cerebral malaria patients correlated with that of PRBC sequestration. As IgE-containing immune complexes are known to induce local overproduction of tumor necrosis factor-alpha (TNF-alpha), a major pathogenic factor in cerebral malaria, IgE may contribute to the pathogenesis of this severe disease. PMID- 11388504 TI - Predisposition to urinary tract epithelial metaplasia in Schistosoma haematobium infection. AB - Although there is strong epidemiologic evidence linking Schistosoma haematobium infection with carcinoma of the bladder, the utility of cytologic screening for urinary tract cancer has not been critically evaluated in S. haematobium-endemic populations. The present cross-sectional study examined urine cytology findings among 1,014 residents (ages 1 to 91) of the S. haematobium-endemic Msambweni area of Coast Province, Kenya. Among 705 evaluable cytology specimens, prevalence of inflammation (39%), hyperkeratosis (30%), metaplasia (33%), and frank atypia (0.4%) was notably higher than in previously studied, non-endemic populations. Overall, S. haematobium infection was strongly associated with increased risk for cytologic abnormality (> 2.8-fold relative risk of metaplasia or hyperkeratosis; P < 0.001). Age-group analysis confirmed parallel increases in metaplasia and S. haematobium infection prevalence early in life (from age I to 15 for both boys and girls). However, above age 20, metaplasia prevalence persisted at 33-45% prevalence despite a decline in infection prevalence and intensity. Prevalence of advanced (moderate or severe) metaplasia showed two age-related peaks: the first at 10-14 years of age (at the time of peak infection), and the second among subjects > or = 60 years old. No cancers were detected in the study population either on cytology or on follow-up ultrasound examination. These data suggest an age-dependent progression of cellular abnormalities in the urinary epithelium that is associated with chronic S. haematobium infection, which becomes independent of concurrent infection intensity as subjects grow older. Implications for cancer screening are discussed. PMID- 11388505 TI - Performance of the OptiMAL assay for detection and identification of malaria infections in asymptomatic residents of Irian Jaya, Indonesia. AB - The OptiMAL assay, a new immunochromatographic "dipstick" test for malaria based on detection of Plasmodium lactate dehydrogenase (pLDH), is purported to detect infections of approximately 200 parasites/microL of blood and to differentiate between Plasmodium falciparum and non-P. falciparum. We evaluated OptiMAL performance by comparing the test strip interpretations of two independent readers with consensus results obtained independently by expert malaria microscopists. Unbiased measures of sensitivity were derived by applying the OptiMAL test for detection and differentiation of light, asymptomatic infections by P. falciparum and Plasmodium vivax. OptiMAL readings were separated in time to determine whether the reaction signal was stable. Microscopy identified infections in 225 of 505 individuals screened; those with P. falciparum (n = 170) averaged 354 asexual forms/microL and P. vivax/Plasmodium malariae (n = 112) averaged 216 asexual forms/microL of blood. Concordance between OptiMAL and microscopy was 81% and 78% by the two independent readings. The assay's sensitivity for detection of any malaria species was 60.4% and 70.2% respectively and specificity was 97% and 89%. Most cases identified by microscopy as P. falciparum were graded as negative or non-falciparum by both OptiMAL readers. OptiMAL false negatives as well as misidentifications were related to low parasitemias (< 500/microL). The OptiMAL assay demonstrated 88-92% sensitivity for detecting infections of 500-1,000 parasites/microL, a range covering the mean parasitemia of primary symptomatic P. falciparum infections in malaria-naive Indonesian transmigrants. This device was markedly less sensitive than expert microscopy for discriminating between malaria species and is presently unsuited for use as an epidemiological screening tool. The OptiMAL assay is not approved for diagnostic use but is commercially available for research purposes only. PMID- 11388506 TI - Comparison of the polymerase chain reaction and serologic tests for diagnosis of septicemic melioidosis. AB - For diagnosis of melioidosis, we compared polymerase chain reaction (PCR)-based DNA detection and three serologic methods with the culture method currently used as gold standard. The diagnostic values of the serologic methods were evaluated in 130 patients. All these patients resided in an endemic area. An enzyme-linked immunosorbent assay (ELISA) gave slightly higher specificity (86.2%) than a dot immunoassay (DOT) (85.3%), but was superior to an indirect hemagglutination assay (IHA) (79.8%). The sensitivities of the DOT (85.7%) and ELISA (71.4%) were considerably higher than that of IHA (61.9%). However, the PCR was the most sensitive (95.2%) and specific (91.7%). Nevertheless, DOT and ELISA are more practical for local hospitals. With the high negative predictive value of both the ELISA (94.0%) and DOT (96.9%) in a high prevalence area, clearly these methods can rule out most of the non-melioidosis patients. PMID- 11388507 TI - Diagnosis of malaria in non-endemic countries by the ParaSight-F test. AB - QBC, examination of thin blood smears, and Parasight-F were performed for every case of malaria suspected between May 1997 and December 1998. Data from 310 patients were reported. Fifty had malaria infection diagnosed by QBC and thin blood film, among whom 39 had Plasmodium falciparum infection. Three of these 39 were negative with the Parasight-F test. Eleven patients had a positive ParaSight F test but microscopic diagnosis methods were negative. Interpretation of these 11 positive ParaSight-F results is proposed to depend on clinical criteria. PMID- 11388508 TI - Use of the recombinant K39 dipstick test and the direct agglutination test in a setting endemic for visceral leishmaniasis in Nepal. AB - We evaluated the field use of two serologic tests for visceral leishmaniasis (VL), the direct agglutination test (DAT) and rK39 dipstick test, in the context of a case-control study. Most VL cases in Nepal are currently diagnosed on clinical grounds and with relatively non-specific tests such as the formol-gel test. Among 14 newly diagnosed VL patients with bone-marrow slides confirmed positive in two independent laboratories, the sensitivity of both tests was 100%. Among 113 controls with no personal or household history of VL, the specificity of the rK39 was 100% while that of the DAT was 93%. The rK39 was less expensive than DAT, and has the advantages of ease of use and obtaining results within minutes. The wider use of the rK39 dipstick test could improve the specificity of VL diagnosis in Nepal. PMID- 11388509 TI - Placental changes associated with fetal outcome in the Plasmodium coatneyi/rhesus monkey model of malaria in pregnancy. AB - Term placentas collected surgically from seven Plasmodium coatneyi-infected rhesus monkeys, one abortion, and five controls were evaluated histopathologically. The placentas from Plasmodium-infected dams had more significant pathologic changes than those from controls for six parameters (P < 0.05) and higher numbers of activated (LN5 + Zymed) macrophages in the intervillous space (IVS) (P = 0.0173). Total parasite load (TPL) was defined as the sum of all weekly peripheral infected red blood cell counts for each trimester and for the entire pregnancy. High first trimester PLs were more likely to result in fetal demise (P = 0.0476) or increased placental damage in surviving infants. As trimester 2-3 TPL increased, so did the number of activated macrophages (P < 0.05) and the total malaria pigment scores (P < 0.05). Low birth weight (LBW) and intrauterine growth retardation (IUGR) were associated with high pigment scores and high numbers of activated macrophages in the IVS. High placental damage scores were not associated with IUGR, LBW, or early infant mortality. PMID- 11388510 TI - Recovery of avian schistosome cercariae from water using penetration stimulant matrix with an unsaturated fatty acid. AB - Avian schistosome cercariae that emerge from aquatic snails can penetrate human skin causing cercarial dermatitis resulting in serious skin disease in sensitized and immunocompromised people. A trap developed for Schistosoma mansoni cercariae was tested for recovery of avian schistosome cercariae. A matrix with an unsaturated fatty acid, linoleic acid stimulates attachment and penetration of Trichobilharzia spp. cercariae, and the immobilized larvae can be subsequently visualized. The number of trapped cercariae exceeded by 3 to 7 times the number of larvae expected on the surface of the trap, based on their random distribution in the water. Recognition, attachment, and penetration of Trichobilharzia spp. cercariae led to injection of more secretory products into the stimulant matrix than by Schistosoma mansoni cercariae. This method can assist in the identification of waters infected with avian schistosome cercariae so that human exposure to these parasitic larvae can be minimized. PMID- 11388512 TI - Factors associated with visceral leishmaniasis in Nepal: bed-net use is strongly protective. AB - Since 1980, visceral leishmaniasis (VL) has reemerged as a public health problem in lowland Nepal. We conducted a case-control study to identify risk factors. In univariate analyses among 84 cases and 105 controls, protective factors included sleeping on a bed or cot (Odds ratio [OR] 0.44, P < 0.01) and sleeping under a bed-net regularly (OR 0.23, P < 0.001) or in the warm months (OR 0.20, P < 0.001). The bed-nets in use in this region were commercially available and untreated with insecticide. Ownership of a cow or buffalo was protective (OR 0.34, P < 0.001), whereas dampness observed in the mud floor of the house was a strong risk factor (OR 4.0, P < 0.001). In multivariable models, bed-net usage, cow or buffalo ownership, and damp floors were significantly associated with altered risk. A program to increase bed-net usage could therefore decrease the incidence of VL in Nepal. PMID- 11388511 TI - Mechanical transport and transmission of Cryptosporidium parvum oocysts by wild filth flies. AB - Over the course of six months wild filth flies were collected from traps left for 7-10 days in a barn with or without a calf shedding Cryptosporidium parvum Genotype 2 oocysts in diarrheic feces. The oocysts of C. parvum transported on the flies' exoskeletons and eluted from their droplets left on visited surfaces were infectious for mice. The mean number of oocysts carried by a fly varied from 4 to 131, and the total oocyst number per collection varied from 56 to approximately 4.56 x 10(3). Fly abundance and intensity of mechanical transmission of infectious C. parvum oocysts were positively correlated, and both increased significantly when an infected calf was in the barn. Molecular data showed that the oocysts shed by infected calves were carried by flies for at least 3 weeks. Filth flies can acquire infectious C. parvum oocysts from unsanitary sites, deposit them on visited surfaces, and therefore may be involved in human or animal cryptosporidiosis. PMID- 11388513 TI - Association between seropositivity of antibodies against hepatitis a virus and Helicobacter pylori. AB - Helicobacter pylori and hepatitis A virus (HAV) are documented to share common transmission routes including fecal-oral. This study examined the association between seropositivity of antibodies against H. pylori (anti-HP) and HAV (anti HAV) via a community-based survey of 40 randomly selected kindergartens in 10 urban and 10 rural areas. Serum samples from 2,047 healthy preschool children and 104 teachers were screened for anti-HP by enzyme-linked immunosorbent assay, and for anti-HAV by microparticle enzyme immunoassay. In children, a low prevalence of anti-HAV (0.44%) was found, in contrast to a high prevalence in their teachers (78.8%); anti-HP seroprevalence was 6.4% for children and 30.8% for teachers. Anti-HAV and anti-HP seropositivities were significantly associated in teachers after adjustment for age, sex, and residential area through multiple logistic regression analysis (multivariate-adjusted odds ratio = 7.3; 95% confidence interval [CI] = 1.4-36.8, P < 0.001). Our findings suggest that HAV and H. pylori may have shared transmission routes in central Taiwan 15 years or more ago, but not any recently. PMID- 11388515 TI - Dynamics of P. falciparum gametocytemia in symptomatic patients in an area of intense perennial transmission in Tanzania. AB - We investigated the dynamics of Plasmodium falciparum gametocytemia in symptomatic patients attending a local dispensary in the Kilombero district, Tanzania. Consenting individuals aged one and above, with varying asexual and sexual parasitemias were treated appropriately and asked to return weekly for 28 days. Gametocyte prevalence was highest on Day 7 of follow-up in all age groups (overall 30.5%). Multifactorial analysis showed that young age (chi2 = 18.4; P = 0.004), high asexual parasitemia on presentation (chi2 = 19.4; P = 0.0007) and gametocyte positivity on presentation (chi2 = 29.4; P = 0.001) were all significantly associated with the presence of gametocytes on Days 7 and 14 of follow-up. High presentation of asexual parasitemia alone was positively correlated with higher gametocyte densities on both days of follow-up (F4, 297 = 2.0; P = 0.049). Gametocyte incidence rates decreased significantly with age (chi2 = 7.6, P < 0.005). In summary, in this group of chloroquine-treated individuals, gametocyte prevalence and incidence rates decreased with age, while densities remained relatively constant. PMID- 11388514 TI - Prevalence and genetic variants of hepatitis GB-C/HG and TT viruses in Gabon, equatorial Africa. AB - The distribution of Hepatitis GB-C/HG (GB-C/HG) and TT viruses (TTV) infections was investigated in selected populations from Gabon using Polymerase Chain Reaction (PCR) and Enzyme Linked Immunosorbent Assay (ELISA) for anti-Envelop 2 (anti-E2) GBV-C/HGV antibodies. Among pregnant women, 29 of 229 (12.6%) were Hepatitis GB virus-C and Hepatitis G virus (GBV-C/HGV) RNA positive (+) and 32 of 81 (39.5%) anti-E2 + versus 8 of 39 (20.5%) TTV DNA +. Among sickle cell anemia patients, 9.7% (3/31) were GBV-C/HGV RNA + versus 22.5% (7/31) TTV DNA +. For tuberculosis patients, the figures were 11.5% (4/35) and 0%. A study of hepatocellular carcinoma cases (n = 27) versus controls (n = 66) did not show significant differences for GBV-C/HGV RNA (10.7% versus 12.1%) and TTV DNA (44.4% versus 30.3%). According to phylogenetic analysis, the 15 GBV-C/HGV strains investigated clustered in group 1, the most common in sub-Saharan Africa whereas TTV sequences (n = 4) mostly clustered in genotypes G1 and one close to genotype G3. In the Gabonese populations investigated, GBV-C/HGV and TTV infections were highly endemic. These data are consistent with the low pathogenicity of these agents. PMID- 11388516 TI - Risk factors associated with leptospirosis in northeastern Thailand, 1998. AB - Leptospirosis is a zoonotic disease of worldwide distribution caused by spirochetes of the genus Leptospira. Humans are infected through direct contact with infected animals or through exposure to fresh water or soil contaminated by infected animal urine. Leptospirosis is characterized by acute fever that can be followed by a more severe, sometimes fatal illness that may include jaundice and renal failure (Weil's disease), meningitis, myocarditis, hemorrhagic pneumonitis, or hemodynamic collapse. To identify potential risk factors for leptospirosis in Thailand, we conducted a matched case-control study in Nakornratchasrima Province of the northeastern region. Fifty-nine cases and 118 controls were included in the study. Four activities in the two weeks prior to illness were independently associated with leptospirosis infection: walking through water (odds ratio [OR] = 4.9, 95% confidence interval [CI] = 1.7-14.1), applying fertilizer in wet fields for more than 6 hr a day (OR = 3.4, 95% CI = 1.5-7.8), plowing in wet fields for more than 6 hr a day (OR = 3.5, 95% CI = 1.1-11.6), and pulling out rice plant sprouts in wet fields for more than 6 hr a day (OR = 3.1, 95% CI = 1.02-9.3). Identification of these risk factors on admission might prove useful for early diagnosis and treatment of leptospirosis in Thailand. PMID- 11388517 TI - An outbreak of fulminant hepatitis delta in the Waorani, an indigenous people of the Amazon basin of Ecuador. AB - An outbreak of delta hepatitis occurred during 1998 among the Waorani of the Amazon basin of Ecuador. Among 58 people identified with jaundice, 79% lived in four of 22 Waorani communities. Serum hepatitis B surface antigen (HBsAg) was found in the sera of 54% of the jaundiced persons, and 14% of asymptomatic persons. Ninety-five percent of 105 asymptomatic Waorani had hepatitis B core (HBc) IgG antibody, versus 98% of 51 with jaundice. These data confirm that hepatitis B virus (HBV) infection is highly endemic among the Waorani. Sixteen of 23 (70%) HBsAg carriers identified at the onset of the epidemic had serologic markers for hepatitis D virus (HDV) infection. All 16 were jaundiced, where as only two of seven (29%) with negative HDV serology were jaundiced (P = .0006). The delta cases clustered in families, 69% were children and most involved superinfection of people chronically infected with HBV. The data suggest that HDV spread rapidly by a horizontal mode of transmission other than by the sexual route. PMID- 11388518 TI - Anesthesiology's greatest generation? PMID- 11388519 TI - Rapacuronium and bronchospasm. PMID- 11388520 TI - A comparison of tracheal tube tip designs on the passage of an endotracheal tube during oral fiberoptic intubation. AB - BACKGROUND: The design of an endotracheal tube has been shown to influence the passage of the tube through the glottis during fiberoptic intubation. Difficulty in passing the endotracheal tube can occur if the aryepiglottic folds obstruct the passage of the bevel. The relevant aspects of endotracheal tube design include the shape of the bevel, the material used by the manufacturer, and the ability of the tube to conform to the shape of the fiberscope. The aim of the current study was to compare the ease of passage through the glottis of two different tubes. One tube was a wire reinforced polyvinyl chloride tube with a standard bevel and the other was a newly designed tube with a bevel of different shape and made of silicone rubber. The new design is for use with the a commerical intubating laryngeal mask. METHODS: The authors studied a population of 30 patients who received a standard anesthetic. In all cases, oral fiberoptic intubation was attempted. Anesthetic was administered to each patient using both tubes, and before the study the order of the tubes was randomized. The difficulty in passing the tube was assessed by a blinded observer and graded using a three point scale (grade 1: no difficulty passing the tube; grade 2: obstruction to passing the tube relieved by withdrawal and a 90 degrees anticlockwise rotation; grade 3: obstruction necessitating more than one manipulation or external laryngeal manipulation). RESULTS: In 27 patients, no difficulty was shown by use of the silicone-tipped tube. In only three patients was there difficulty that necessitated a 90 degrees anticlockwise twist. With the wire-reinforced tube, no difficulty was experienced on 14 occasions. Grade 1 difficulty was experienced eight times and difficulty necessitating more than one maneuver, head movement, or external laryngeal manipulation was seen on eight occasions. Statistical significance was achieved at P = 0.0002 (Wilcoxon signed rank test). CONCLUSIONS: The authors conclude that the use of the silicone-tipped tube with the new bevel design may provide an advantage in the clinical situation of fiberoptic intubation. PMID- 11388521 TI - Dose-dependent regional cerebral blood flow changes during remifentanil infusion in humans: a positron emission tomography study. AB - BACKGROUND: The current study investigated dose-dependent effects of the mu selective agonist remifentanil on regional cerebral blood flow (rCBF) in volunteers using positron emission tomography (PET). METHODS: Ten right-handed male volunteers were included in a 15O-water PET study. Seven underwent three conditions: control (saline), low remifentanil (0.05 microg x kg(-1) x min(-1)), and moderate remifentanil (0.15 microg x kg(-1) x min(-1)). The remaining three participated in the low and moderate conditions. A semirandomized study protocol was used with control and remifentanil conditions 3 or more months apart. The order of low and moderate conditions was randomized. Cardiovascular and respiratory parameters were monitored. Categoric comparisons between the control, low, and moderate conditions and a pixelwise correlation analysis across the three conditions were performed (P < 0.05, corrected for multiple comparisons) using statistical parametric mapping. RESULTS: Cardiorespiratory parameters were maintained constant over time. At the low remifentanil dose, significant increases in relative rCBF were noted in the lateral prefrontal cortices, inferior parietal cortices, and supplementary motor area. Relative rCBF decreases were observed in the basal mediofrontal cortex, cerebellum, superior temporal lobe, and midbrain gray matter. Moderate doses further increased rCBF in mediofrontal and anterior cingulate cortices, occipital lobe transition, and caudal periventricular grey. Significant decreases were detected in the inferior parietal lobes. These dose-dependent effects of remifentanil on rCBF were confirmed by a correlation analysis. CONCLUSION: Remifentanil induced dose dependent changes in relative rCBF in areas involved in pain processing. At moderate doses, rCBF responses were additionally detected in structures known to participate in modulation of vigilance and alertness. Insight into the mechanisms of opioid analgesia within the pain-processing neural network may lead to a better understanding of antinociception and opioid treatment. PMID- 11388522 TI - Minimum analgesic dose of epidural sufentanil for first-stage labor analgesia: a comparison between spontaneous and prostaglandin-induced labors in nulliparous women. AB - BACKGROUND: The aim of this prospective, double-blind, sequential allocation study was to compare the effects of spontaneous and prostaglandin-induced labor on the minimum analgesic dose of epidural sufentanil in the first stage of labor. METHODS: Seventy healthy, nulliparous women, at more than 37 weeks' gestation with cervical dilatation from 2 to 4 cm, requesting epidural pain relief in labor were enrolled. The subjects were assigned to two different groups according to whether labor was spontaneous or induced with dinoprostone 0.5 mg. Parturients received 10 ml of the study solution through a lumbar epidural catheter. The initial dose was sufentanil 25 microg, and subsequent doses were determined by the response of the previous patient in the same group using up-down sequential allocation. The analgesic effectiveness was assessed using 100-mm visual analog pain scores. The up-down sequences were analyzed using the method of independent paired reversals and probit regression. RESULTS: The minimum analgesic dose of sufentanil in spontaneous labor was 22.2 microg (95% CI: 19.6, 22.8) and 27.3 microg (95% CI: 23.8, 30.9) in induced labor. The minimum analgesic dose of sufentanil in induced labor was significantly greater (P = 0.0014) than that in spontaneous labor (95% CI difference: 2.9, 9.3) by a factor of 1.3 (95% CI: 1.1, 1.5). CONCLUSION: Prostaglandin induction of labor produces a significantly greater analgesic requirement than does spontaneous labor. PMID- 11388523 TI - Tissue injury and the inflammatory response to pediatric cardiac surgery with cardiopulmonary bypass: a descriptive study. AB - BACKGROUND: There are few detailed descriptions of the inflammatory response to cardiac surgery with cardiopulmonary bypass (CPB) in children beyond 24 h postoperatively. This is especially true for the antiinflammatory cytokines and the extent of tissue injury. The aim of the current study was to describe the inflammatory and injury responses in uncomplicated pediatric cardiac surgery with CPB, where methylprednisolone and modified ultrafiltration (MUF) were used. METHODS: Blood samples were collected up to 48 h postoperatively. Cytokines (tumor necrosis factor-alpha and interleukin-6, -1beta, -10, and -1ra), complement (C3d and C4d) and coagulation system (prothrombin activation fragments 1 and 2 and antithrombin III) activation, neutrophil elastase, and the resulting tissue injury (creatine kinase, lactate dehydrogenase, alanine transaminase, amylase, and gamma-glutamyl transferase) were measured. RESULTS: The proinflammatory cytokine release varied widely, in contrast to a clear-cut antiinflammatory response. Cytokine concentrations did not decrease immediately after MUF, and no rebound increases later in the postoperative period were observed. The coagulation system, but not complement, was activated. There was a late release of C-reactive protein. Tissue injury could be quantified biochemically without evidence of hepatic or pancreatic dysfunction. CONCLUSION: In this group of uncomplicated subjects, the antiinflammatory cytokine and tissue injury responses were well defined, in contrast to a variable proinflammatory cytokine release. This was accompanied by activation of the coagulation system but not of complement. Concentrations of inflammatory mediators did not decrease immediately after MUF, and there was no evidence for rebound release later in the postoperative period. PMID- 11388524 TI - Hemoglobin desaturation after succinylcholine-induced apnea: a study of the recovery of spontaneous ventilation in healthy volunteers. AB - BACKGROUND: Because of the rapid recovery of neuromuscular function after succinylcholine administration, there is a belief that patients will start breathing sufficiently rapidly to prevent significant oxygen desaturation. The authors tested whether this belief was valid. METHODS: Twelve healthy volunteers aged 18-45 yr participated in the study. After preoxygenation to an end-tidal oxygen concentration greater than 90%, each subject received 5 mg/kg thiopental and 1 mg/kg succinylcholine. Oxygen saturation (SaO2) was measured at both a finger and an ear lobe (beat to beat). During the period of apnea and as they were recovering, the volunteers received continuous verbal reassurance by the investigators. If the SaO2 decreased below 80%, the volunteers received chin lift and, if necessary, assisted ventilation. The length of time the subject was apneic and level of desaturation were related by linear regression analysis. One hour after recovery and again 1 week later, subjects were asked a series of questions regarding their emotional experience. RESULTS: In six volunteers, SaO2 decreased below 95% during apnea; in four, SaO2 decreased below 80%, necessitating chin lift and assisted ventilation in three. Apnea time was significantly longer in volunteers who reached SaO2 less than 80% than in those who did not (7.0+/-0.4 and 4.1+/-0.3 min, respectively), and there was a significant correlation between the length of time the subject was apneic and the magnitude of desaturation. CONCLUSIONS: Spontaneous recovery from succinylcholine induced apnea may not occur sufficiently quickly to prevent hemoglobin desaturation in subjects whose ventilation is not assisted. PMID- 11388525 TI - Pharyngolaryngeal morbidity with the laryngeal mask airway in spontaneously breathing patients: does size matter? AB - BACKGROUND: Currently, the manufacturer of the laryngeal mask airway (LMA; Laryngeal Mask Company, Ltd., Northfield End, Henley on Thames, Oxon, United Kingdom) recommends using as large a mask size as possible. The aim of this study was to compare the incidence of pharyngolaryngeal morbidity after the use of a large (size 5 in males and size 4 in females) or small (size 4 in males and size 3 in females) LMA in spontaneously breathing patients. METHODS: A total of 258 male and female patients were randomly assigned to insertion of a large or small LMA while breathing spontaneously during general anesthesia. After insertion of the LMA, a "just-seal" cuff pressure was obtained, and intracuff pressure was measured at 10-min intervals until just before removal of the LMA. The 2- and 24 h incidence of postoperative sore throat, pain, hoarseness, dysphagia, and nausea and vomiting was assessed. Complications after LMA removal, including body movement, coughing, retching, regurgitation, vomiting, biting on the LMA, bronchospasm, laryngospasm, or the presence of blood on the LMA, were recorded. RESULTS: The use of a large LMA was associated with a higher incidence of sore throat in both sexes (20% vs. 7% in men, 21% vs. 5% in women; P < 0.05) and a higher incidence of hoarseness in male patients at 2 h postoperatively (21% vs. 9%, P < 0.05). There was a higher incidence of sore throat in male patients at 24 h postoperatively with the use of a large LMA (26% vs. 12%, P < 0.05). There was no difference in the incidence of complications of LMA removal orother pharyngolaryngeal morbidity, such as difficulty swallowing, drinking, and eating, or nausea and vomiting, between male or female groups at any time period with the use of a large LMA. CONCLUSIONS: Selection of a small laryngeal mask airway (size 4) in spontaneously breathing male patients may be more appropriate to limit the occurrence of sore throat on the first postoperative day. All patients had a fourfold increased risk of developing sore throat when a large LMA was used. PMID- 11388526 TI - Dose-response study of epidural ropivacaine for labor analgesia. AB - BACKGROUND: Ropivacaine has been introduced for use in epidural analgesia in labor. However, there have been few formal dose-response studies of ropivacaine in this setting. METHODS: The authors performed a prospective, randomized, double blind study examining the effectiveness of five different doses of ropivacaine (10, 20, 30, 40, and 50 mg) administered epidurally in a volume of 10 ml to establish analgesia in 66 parturients who were in active labor with cervical dilatation less than 4 cm. A dose was considered effective when the visual analog scale pain score decreased by 50% or more from baseline. RESULTS: A sigmoid dose response curve and a probit log dose-response plot (linear regression coefficient, r = 0.84; coefficient of determination, r2 = 0.71) were obtained. The ED50 (median effective dose) obtained based on the maximum likelihood estimation was 18.4 mg (95% confidence interval, 13.4-25.4 mg). Time to onset of analgesia, duration of analgesia, time to two-segment regression of sensory block level, and incidence of motor block were not affected by the dosage of ropivacaine administered (P = 0.93, 0.12, 0.55, and 0.39, respectively). However, the upper level of sensory block was dose-related (P < 0.01). CONCLUSION: In a traditional dose-response study, the ED50 of ropivacaine required to initiate epidural analgesia in early labor was found to be 18.4 mg (95% confidence interval, 13.4-25.4 mg). PMID- 11388527 TI - Efficacy of a simple intraoperative transfusion algorithm for nonerythrocyte component utilization after cardiopulmonary bypass. AB - BACKGROUND: Abnormal bleeding after cardiopulmonary bypass (CPB) is a common complication of cardiac surgery, with important health and economic consequences. Coagulation test-based algorithms may reduce transfusion of non-erythrocyte allogeneic blood in patients with abnormal bleeding. METHODS: The authors performed a randomized prospective trial comparing allogeneic transfusion practices in 92 adult patients with abnormal bleeding after CPB. Patients with abnormal bleeding were randomized to one of two groups: a control group following individual anesthesiologist's transfusion practices and a protocol group using a transfusion algorithm guided by coagulation tests. RESULTS: Among 836 eligible patients having all types of elective cardiac surgery requiring CPB, 92 patients developed abnormal bleeding after CPB (incidence, 11%). The transfusion algorithm group received less allogeneic fresh frozen plasma in the operating room after CPB (median, 0 units; range, 0-7 units) than the control group (median, 3 units; range, 0-10 units) (P = 0.0002). The median number of platelet units transfused in the operating room after CPB was 4 (range, 0-12) in the algorithm group compared with 6 (range, 0-18) in the control group (P = 0.0001). Intensive care unit (ICU) mediastinal blood loss was significantly less in the algorithm group. Multivariate analysis demonstrated that transfusion algorithm use resulted in reduced ICU blood loss. The control group also had a significantly greater incidence of surgical reoperation of the mediastinum for bleeding (11.8% vs. 0%; P = 0.032). CONCLUSIONS: Use of a coagulation test-based transfusion algorithm in cardiac surgery patients with abnormal bleeding after CPB reduced non-erythrocyte allogeneic transfusions in the operating room and ICU blood loss. PMID- 11388529 TI - Dose-response relationship and infusion requirement of cisatracurium besylate in infants and children during nitrous oxide-narcotic anesthesia. AB - BACKGROUND: To determine the effect of age on the dose-response relation and infusion requirement of cisatracurium besylate in pediatric patients, 32 infants (mean age, 0.7 yr; range, 0.3-1.0 yr) and 32 children (mean age, 4.9 yr; range, 3.1-9.6 yr) were studied during thiopentone-nitrous oxideoxygen-narcotic anesthesia. METHODS: Potency was determined using a single-dose (20, 26, 33, or 40 microg/kg) technique. Neuromuscular block was assessed by monitoring the electromyographic response of the adductor pollicis to supramaximal train-of-four stimulation of the ulnar nerve at 2 Hz. RESULTS: Least-squares linear regression analysis of the log-probit transformation of dose and maximal response yielded median effective dose (ED50) and 95% effective dose (ED95) values for infants (29+/-3 microg/kg and 43+/-9 microg/kg, respectively) that were similar to those for children (29+/-2 microg/kg and 47+/-7 microg/kg, respectively). The mean infusion rate necessary to maintain 90-99% neuromuscular block during the first hour in infants (1.9+/-0.4 microg x kg(-1) x min(-1); range: 1.3-2.5 microg x kg( 1) x min(-1)) was similar to that in children (2.0+/-0.5 microg x kg(-1) x min( 1); range: 1.3-2.9 microg x kg(-1) x min(-1)). CONCLUSION: The authors conclude that cisatracurium is equipotent in infants and children when dose is referenced to body weight during balanced anesthesia. PMID- 11388528 TI - The midlatency auditory evoked potentials predict responsiveness to verbal commands in patients emerging from anesthesia with xenon, isoflurane, and sevoflurane but not with nitrous oxide. AB - BACKGROUND: It has recently been demonstrated that the approximately 40-Hz spectral power of the midlatency auditory evoked potential (MLAEP) correlates well with wakefulness during desflurane or propofol anesthesia. The aim of this study was to characterize how other inhalational anesthetics affects the MLAEP as the patients regain responsiveness to simple verbal command during emergence from anesthesia. METHODS: Sixty patients were randomly assigned to receive xenon, isoflurane, sevoflurane, or nitrous oxide (N2O) supplemented with epidural anesthesia. During emergence, the concentration of an anesthetic was decreased in 0.1-minimum alveolar concentration (MAC) decrements from 0.8 MAC or from 70% in the case of N2O, and each new concentration was maintained for 15 min. Every 5 min during each equilibration period, the MLAEP was recorded and the patients were asked to open their eyes and squeeze and release the investigator's hand. This process was repeated until the first response to either of these commands was observed. RESULTS: Thirteen patients were excluded because of technical reasons. The preanesthetic MLAEP showed a periodic waveform, where the Na-Pa-Nb complex was the most prominent component contributing to the high energy around 29-39 Hz in the power spectrum. Emergence from xenon, isoflurane, and sevoflurane anesthesia produced similar changes in the MLAEP. The spectral power for the frequency 29 Hz or greater was severely suppressed at 0.8 MAC but significantly recovered between the concentration only 0.1 MAC higher that permitting the first response to command and that associated with the first response. In contrast, N2O hardly affected the MLAEPs, even at the concentrations producing unresponsiveness. Two patients did not lose responsiveness even at the highest concentration tested (70%). CONCLUSIONS: The MLAEP is closely associated with responsiveness to verbal command during emergence from anesthesia with xenon, isoflurane, and sevoflurane but not with N2O. PMID- 11388530 TI - Density detection in dependent left lung region using transesophageal echocardiography. AB - BACKGROUND: Densities in dependent lung regions worsen oxygenation in patients with acute respiratory distress syndrome. Identification of these densities requires examination using computed tomography (CT). In this study, the authors evaluated the use of transesophageal echocardiography (TEE) to estimate densities in the dependent lung. METHODS: Forty consecutive patients with acute lung injury or acute respiratory distress syndrome who underwent CT and TEE examination were included in this study. Densities in the lower left lung area were detected through the descending aorta by TEE. Density areas observed by TEE were compared with those obtained by CT. The effect of positive end-expiratory pressure (PEEP) application on density area was also evaluated. RESULTS: Density areas in the dependent lung region measured by TEE were 12.0+/-6.1 cm2 (mean +/- SD) at mid esophageal position. Density areas evaluated using TEE in the left lung correlated significantly with those estimated with CT in the left and right lungs (P < 0.01 in both lungs). In addition, the authors observed a significant correlation between PaO2/FIO2 and density areas estimated using TEE (P < 0.05). During positive end-expiratory pressure application, the area of density estimated with TEE decreased and PaO2 improved. CONCLUSIONS: The authors clearly demonstrated that it is possible to estimate the density area of the dependent left lung regions in patients with acute lung injury or acute respiratory distress syndrome using TEE. It is also possible to observe the changes of density areas during application of positive end-expiratory pressure. PMID- 11388531 TI - Epidural lidocaine decreases sevoflurane requirement for adequate depth of anesthesia as measured by the Bispectral Index monitor. AB - BACKGROUND: Epidural anesthesia potentiates sedative drug effects and decreases minimum alveolar concentration (MAC). The authors hypothesized that epidural anesthesia also decreases the general anesthetic requirements for adequate depth of anesthesia as measured by Bispectral Index (BIS). METHODS: After premedication with 0.02 mg/kg midazolam and 1 microg/kg fentanyl, 30 patients aged 20-65 yr were randomized in a double-blinded fashion to receive general anesthesia with either intravenous saline placebo or intravenous lidocaine control (1-mg/kg bolus dose; 25 microg x kg(-1) x min(-1)). A matched group was prospectively assigned to receive epidural lidocaine (15 ml; 2%) with intravenous saline placebo. All patients received 4 mg/kg thiopental and 1 mg/kg rocuronium for tracheal intubation. After 10 min of a predetermined end-tidal sevoflurane concentration, BIS was measured. The ED50 of sevoflurane for each group was determined by up down methodology based on BIS less than 50 (MAC(BIS50)). Plasma lidocaine concentrations were measured. RESULTS: The MAC(BIS50) of sevoflurane (0.59% end tidal) was significantly decreased with lidocaine epidural anesthesia compared with general anesthesia alone (0.92%) or with intravenous lidocaine (1%; P < 0.0001). Plasma lidocaine concentrations in the intravenous lidocaine group (1.9 microg/ml) were similar to those in the epidural lidocaine group (2.0 microg/ml). CONCLUSIONS: Epidural anesthesia reduced by 34% the sevoflurane required for adequate depth of anesthesia. This effect was not a result of systemic lidocaine absorbtion, but may have been caused by deafferentation by epidural anesthesia or direct rostral spread of local anesthetic within the cerebrospinal fluid. Lower than-expected concentrations of volatile agents may be sufficient during combined epidural-general anesthesia. PMID- 11388532 TI - Interaction of edrophonium with muscarinic acetylcholine M2 and M3 receptors. AB - BACKGROUND: It has been reported that edrophonium can antagonize the negative chronotropic effect of carbachol. This study was undertaken to evaluate in detail the interaction of edrophonium with muscarinic Mz and M3 receptors. METHODS: A functional study was conducted to evaluate the effects of edrophonium on the concentration-response curves for the negative chronotropic effect and the bronchoconstricting effect of carbachol in spontaneously beating right atria and tracheas of guinea pigs. An electrophysiologic study was conducted to compare the effects of edrophonium on carbachol-, guanosine triphosphate (GTP)gama S-, and adenosine-induced outward K+ currents in guinea pig atrial cells by whole cell voltage clamp technique. A radioligand binding study was conducted to examine the effects of edrophonium on specific [3HIN-methylscopolamine (NMS) binding to guinea pig atrial (M2) and submandibular gland (M3) membrane preparations, and on atropine-induced dissociation of [3H]NMS. RESULTS: Edrophonium shifted rightward the concentration-response curves for the negative chronotropic and bronchoconstricting effects of carbachol in a competitive manner. The pA2 values for cardiac and tracheal muscarinic receptors were 4.61 and 4.03, respectively. Edrophonium abolished the carbachol-induced outward current without affecting the GTPgamma S- and adenosine-induced currents in the atrial cells. Edrophonium inhibited [3H]NMS binding to M2 and M3 receptors in a concentration-dependent manner. The pseudo-Hill coefficient values and apparent dissociation constants of edrophonium for M2 and M3 receptors were 1.02 and 1.07 and 21 and 34 microM, respectively. Edrophonium also changed dissociation constant values of [3H]NMS without affecting its maximum binding capacities. CONCLUSION: Edrophonium binds to muscarinic M2 and M3 receptors nonselectively, and acts as a competitive antagonist. PMID- 11388533 TI - Pulmonary vascular effects of propofol at baseline, during elevated vasomotor tone, and in response to sympathetic alpha- and beta-adrenoreceptor activation. AB - BACKGROUND: This in vivo study had two primary objectives. The first goal was to determine whether the pulmonary vascular effects of propofol depend on the preexisting level of vasomotor tone, and the second was to investigate the effects of propofol on the pulmonary vascular responses to sympathetic alpha- and beta-adrenoreceptor activation. METHODS: Thirty-one mongrel dogs were chronically instrumented to measure the left pulmonary vascular pressure-flow (LPQ) relation. Left lung autotransplantation (LLA) was also performed in eight additional dogs to induce a long-term increase in pulmonary vascular resistance. LPQ plots were measured on separate days in the conscious state and during propofol anesthesia. LPQ plots were measured at baseline and when vasomotor tone was acutely increased with the alpha agonist, phenylephrine, or the thromboxane mimetic, U46619. In separate experiments, cumulative dose-response curves to alpha- (phenylephrine) and beta- (isoproterenol) adrenoreceptor agonists were generated in conscious and propofol-anesthetized dogs. RESULTS: Compared with the conscious state, propofol had no effect on the baseline LPQ relation in normal or post-LLA dogs. However, propofol caused pulmonary vasoconstriction (P < 0.05) when vasomotor tone was acutely increased with either phenylephrine or U46619 in normal or post-LLA dogs. The pulmonary vasoconstrictor response to alpha-adrenoreceptor activation was potentiated (P < 0.05) during propofol anesthesia, whereas the pulmonary vasodilator response to beta-adrenoreceptor activation was not altered. CONCLUSION: These results indicate that the pulmonary vascular response to propofol anesthesia is tone-dependent. During sympathetic activation, propofol may favor alpha-adrenoreceptor-mediated vasoconstriction over beta-adrenoreceptor mediated vasodilation. PMID- 11388534 TI - Anesthetic potency and influence of morphine and sevoflurane on respiration in mu opioid receptor knockout mice. AB - BACKGROUND: The involvement of the mu-opioid receptor (muOR) system in the control of breathing, anesthetic potency, and morphine- and anesthesia-induced respiratory depression was investigated in mice lacking the muOR. METHODS: Experiments were performed in mice lacking exon 2 of the muOR gene (muOR-/-) and their wild-type littermates (muOR+/+). The influence of saline, morphine, naloxone, and sevoflurane on respiration was measured using a whole body plethysmographic method during air breathing and elevations in inspired carbon dioxide concentration. The influence of morphine and naloxone on anesthetic potency of sevoflurane was determined by tail clamp test. RESULTS: Relative to wild-type mice, muOR-deficient mice displayed approximately 15% higher resting breathing frequencies resulting in greater resting ventilation levels. The slope of the ventilation-carbon dioxide response did not differ between genotypes. In muOR+/+ but not muOR-/- mice, a reduction in resting ventilation and slope, relative to placebo, was observed after 100 mg/kg morphine. Naloxone increased resting ventilation and slope in both genotypes. Sevoflurane at 1% inspired concentration induced similar reductions in resting ventilation and slope in the two genotypes. Anesthetic potency was 20% lower in mutant relevant to wild-type mice. Naloxone and morphine caused an increase and decrease, respectively, in anesthetic potency in muOR+/+ mice only. CONCLUSIONS: The data indicate the importance of the endogenous opioid system in the physiology of the control of breathing with only a minor role for the muOR. The muOR gene is the molecular site of action of the respiratory effects of morphine. Anesthetic potency is modulated by the endogenous mu-opioid system but not by the kappa- and delta opioid systems. PMID- 11388535 TI - Propofol potentiates phenylephrine-induced contraction via cyclooxygenase inhibition in pulmonary artery smooth muscle. AB - BACKGROUND: The authors previously demonstrated in vivo that the pulmonary vasoconstrictor response to the a agonist phenylephrine is potentiated during propofol anesthesia compared with the conscious state. The current in vitro study tested the hypothesis that propofol potentiates phenylephrine-induced contraction by inhibiting the synthesis and/or activity of vasodilator metabolites of the cyclooxygenase pathway. METHODS: Canine pulmonary arterial rings were suspended for isometric tension recording. Intracellular calcium concentration ([Ca2+]i) was measured in pulmonary arterial strips loaded with acetoxylmethyl ester of fura-2. After phenylephrine-induced contraction, propofol (10(-7) to 10(-4) M) was administered in the presence or absence of the cyclooxygenase inhibitor ibuprofen (10(-5) M). The effects of propofol on the arachidonic acid and prostacyclin relaxation-response curves were assessed. The amount of 6-keto prostaglandin F1alpha (stable metabolite of prostacyclin) released from pulmonary vascular smooth muscle in response to phenylephrine was measured with enzyme immunoassay in the presence or absence of propofol and ibuprofen. RESULTS: Propofol potentiated phenylephrine-induced contraction in pulmonary arterial rings in a concentration-dependent and endothelium-independent manner. In endothelium-denuded strips, propofol (10(-4) M) increased tension by 53+/-11%, and increased [Ca2+]i by 56+/-9%. Ibuprofen also potentiated phenylephrine induced contraction but abolished the propofol-induced increases in tension and [Ca2+]i. Propofol had no effect on the relaxation response to prostacyclin, whereas propofol and ibuprofen attenuated the relaxation response to arachidonic acid to a similar extent. Phenylephrine markedly increased 6-keto prostaglandin F1alpha production, and this effect was virtually abolished by propofol and ibuprofen. CONCLUSION: These results suggest that propofol potentiates alpha adrenoreceptor-mediated pulmonary vasoconstriction by inhibiting the concomitant production of prostacyclin by cyclooxygenase. PMID- 11388536 TI - Comparison of volatile anesthetic actions on intracellular calcium stores of vascular smooth muscle: investigation in isolated systemic resistance arteries. AB - BACKGROUND: Volatile anesthetic actions on intracellular Ca2+ stores (ie., sarcoplasmic reticulum [SR]) of vascular smooth muscle have not been fully elucidated. METHODS: Using isometric force recording method and fura-2 fluorometry, the actions of four volatile anesthetics on SR were studied in isolated endothellum-denuded rat mesenteric arteries. RESULTS: Halothane (> or = 3%) and enflurane (> or = 3%), but not isoflurane and sevoflurane, increased the intracellular Ca2+ concentration ([Ca2+]i) in Ca2+-free solution. These Ca2+ releasing actions were eliminated by procaine. When each anesthetic was applied during Ca2+ loading, halothane (> or = 3%) and enflurane (5%), but not isoflurane and sevoflurane, decreased the amount of Ca2+ in the SR. However, if halothane or enflurane was applied with procaine during Ca2+ loading, both anesthetics increased the amount of Ca2+ in the SR. The caffeine-induced increase in [Ca2+], was enhanced in the presence of halothane (> or = 1%), enflurane (> or = 1%), and isoflurane (> or = 3%) but was attenuated in the presence of sevoflurane (> or = 3%). The norepinephrine-induced increase in [Ca2+], was enhanced only in the presence of sevoflurane (> or = 3%). Not all of these anesthetic effects on the [Ca2+]i were parallel with the simultaneously observed anesthetic effects on the force. CONCLUSIONS: In systemic resistance arteries, the halothane, enflurane, isoflurane, and sevoflurane differentially influence the SR functions. Both halothane and enflurane cause Ca2+ release from the caffeine-sensitive SR. In addition, both anesthetics appear to have a stimulating action on Ca2+ uptake in addition to the Ca2+-releasing action. Halothane, enflurane, and isoflurane all enhance, while sevoflurane attenuates, the Ca2+-induced Ca2+-release mechanism. However, only sevoflurane stimulates the inositol 1,4,5-triphosphate-induced Ca2+ release mechanism. Isoflurane and sevoflurane do not stimulate Ca2+ release or influence Ca2+ uptake. PMID- 11388537 TI - Efficacy of propofol to prevent bronchoconstriction: effects of preservative. AB - BACKGROUND: The authors previously showed that propofol attenuates bronchoconstriction. Recently, a newer formulation of propofol with metabisulfite preservative has been introduced. metabisulfite causes airway narrowing in asthmatics. Therefore, we tested whether the preservative metabisulfite abolishes the ability of propofol to attenuate bronchoconstriction. The authors used a sheep model in which anesthetic agents could be directly administered to the airways via the bronchial artery. METHODS: After Internal Review Board approval, seven sheep were anesthetized (pentobarbital 20 mg x kg(-1) x h(-1)) and paralyzed (pancuronium 2 mg), and the lungs were ventilated. After left thoracotomy, the bronchial artery was cannulated and perfused. In random order, propofol with and without metabisulfite, lidocaine (5 mg/ml), or metabisulfite alone (0.125 mg/ml) was infused into the bronchial artery at a rate of 0.06, 0.2, or 0.6 ml/min. After 10 min, airway resistance (Raw) was measured before and after vagal nerve stimulation (30 Hz, 30-ms duration at 30 V for 9 s.) and methacholine challenge (2 microg/ml at 2 ml/min in the bronchial artery). Data were expressed as a percent of maximal response and analyzed by analysis of variance with correction and with significance accepted at P < or = 0.05. RESULTS: Raw at baseline was not significantly different among the four drugs (P = 0.87). Infusion of lidocaine and propofol without metabisulfite into the bronchial artery caused a dose-dependent attenuation of the vagal nerve stimulation-induced bronchoconstriction (P = 0.001). Propofol with metabisulfite had no effect on vagal nerve stimulation-induced bronchoconstriction (P = 0.40). There was a significant difference in the ability of propofol without metabisulfite compared with propofol with metabisulfite to attenuate vagal nerve stimulation-induced (P = 0.0001) and methacholine-induced bronchoconstriction (P = 0.0001). CONCLUSION: Propofol without metabisulfite and lidocaine attenuated vagal nerve stimulation-induced bronchoconstriction in a dose-dependent fashion. Propofol without metabisulfite also decreased direct airway smooth muscle constriction. The preservative used for propofol can have a dramatic effect on its ability to attenuate bronchoconstriction. PMID- 11388538 TI - Effects of isoflurane on intracellular calcium and myocardial crossbridge kinetics in tetanized papillary muscles. AB - BACKGROUND: Isoflurane depresses the intracellular Ca2+ transient and force development during a twitch, but its effects on crossbridge cycling rates are difficult to predict because of the transient nature of the twitch. Measurements of the effects of isoflurane on crossbridge cycling kinetics during tetanic contractions, which provide a steady state level of activation in intact cardiac muscle, have not been previously reported. METHODS: Ferret right ventricular papillary muscles were isolated, and superficial cells were microinjected with the bioluminescent photoprotein aequorin to monitor the intracellular Ca2+ concentration. The rate of tension redevelopment (kTR) was measured during steady state isometric activation (tetanic stimulation, frequency 20 Hz, 1 microM ryanodine, temperature = 30 degrees C) in the absence of isoflurane (2, 6, and 12 mM extracellular [Ca2+]) and in the presence of 0.5, 1.0, and 1.5 minimum alveolar concentration isoflurane (12 mM extracellular [Ca2+]). RESULTS: Intracellular [Ca2+], isometric force, and kTR all increased when the extracellular [Ca2+] increased. Isoflurane (0.5, 1.0, and 1.5 minimum alveolar concentration) caused intracellular [Ca2+], isometric force, and kTR to decrease in a dose-dependent manner in the presence of 12 mM extracellular [Ca2+]. In the presence of increasing concentrations of isoflurane, the relation between intracellular [Ca2+] and force remained unchanged, whereas the relation between intracellular [Ca2+] and kTR was shifted toward higher [Ca2+]. CONCLUSIONS: These results indicate that isoflurane depresses myocardial crossbridge cycling rates. It appears that this effect is partially mediated by a decrease in the intracellular [Ca2+]. However, additional mechanisms must be considered to explain the shift of the relation between intracellular [Ca2+] and kTR toward higher [Ca2+]. PMID- 11388539 TI - Optimal mean airway pressure during high-frequency oscillation: predicted by the pressure-volume curve. AB - BACKGROUND: A number of groups have recommended setting positive end-expiratory pressure during conventional mechanical ventilation in adults at 2 cm H2O above the lower corner pressure (P(CL)) of the inspiratory pressure-volume (P-V) curve of the respiratory system. No equivalent recommendations for the setting of the mean airway pressure (Paw) during high-frequency oscillation (HFO) exist. The authors questioned if the Paw resulting in the best oxygenation without hemodynamic compromise during HFO is related to the static P-V curve in a large animal model of acute respiratory distress syndrome. METHODS: Saline lung lavage was performed in seven sheep (28+/-5 kg, mean +/- SD) until the arterial oxygen partial pressure/fraction of inspired oxygen ratio decreased to 85+/-27 mmHg at a positive end-expiratory pressure of 5 cm H2O (initial injury). The PCL (20+/-1 cm H2O) on the inflation limb and the point of maximum curvature change (PMC; 26+/-1 cm H2O) on the deflation limb of the static P-V curve were determined. The sheep were subjected to four 1-h cycles of HFO at different levels of Paw (P(CL) + 2, + 6, + 10, + 14 cm H2O), applied in random order. Each cycle was preceded by a recruitment maneuver at a sustained Paw of 50 cm H2O for 60 s. RESULTS: High frequency oscillation with a Paw of 6 cm H2O above P(CL) (P(CL) + 6) resulted in a significant improvement in oxygenation (P < 0.01 vs. initial injury). No further improvement in oxygenation was observed with higher Paw, but cardiac output decreased, pulmonary vascular resistance increased, and oxygen delivery decreased at Paw greater than P(CL) + 6. The PMC on the deflation limb of the P-V curve was equal to the P(CL) + 6 (r = 0.77, P < 0.05). CONCLUSION: In this model of acute respiratory distress syndrome, optimal Paw during HFO is equal to P(CL) + 6, which correlates with the PMC. PMID- 11388540 TI - Differential behavioral effects of peripheral and systemic morphine and naloxone in a rat model of repeated acute inflammation. AB - BACKGROUND: It has been reported that opioid antinociceptive effects are enhanced in animal models of inflammation, but it remains unclear whether this sensitization to morphine is related to predominant central or peripheral increased effects. METHODS: The authors compared the behavioral effects of intraplantar and intravenous morphine and naloxone in a rat model of repeated acute carrageenan-induced inflammation in which enhanced responses to noxious stimuli result from sensitization in peripheral tissues or central sensitization. The antinociceptive effects of intraplantar morphine (50, 75, 100, 150, and 200 microg), intravenous morphine (0.3, 0.6, and 1 mg/kg), and the pronociceptive effects of intraplantar naloxone methiodide (150 microg) and intravenous naloxone (1 mg/kg) against noxious pressure (vocalization thresholds to paw pressure) in rats were assessed 3 h after one or two carrageenan plantar injections performed 7 days apart. RESULTS: After the first carrageenan injection, intraplantar and intravenous morphine produced significant increase of vocalization thresholds to paw pressure in inflamed but not in noninflamed paws. After the second carrageenan injection, the antinociceptive effects of intraplantar morphine were significantly reduced compared with those obtained after the first carrageenan injection, whereas effects of intravenous morphine were significantly enhanced and present in both hind paws. Intravenous naloxone demonstrated similar pronociceptive patterns after the first and second carrageenan injection. Intraplantar naloxone methiodide produced pronociceptive effects in inflamed hind paw that were significantly enhanced after the second carrageenan injection. CONCLUSIONS: When inflammation is enhanced by recurrent stimulations, the antinociceptive effects of systemic morphine are enhanced. This increase is more likely related to central than peripheral sites of action, beyond endogenous opioid system activation. PMID- 11388541 TI - Epinephrine increases the neurotoxic potential of intrathecally administered lidocaine in the rat. AB - BACKGROUND: Epinephrine is commonly added to lidocaine solutions to increase the duration of spinal anesthesia. Despite this common usage, the effect of epinephrine on the neurotoxic potential of this anesthetic is not known. The current experiments investigated whether adding epinephrine increases functional impairment or histologic damage induced by spinal administration of lidocaine in the rat. METHODS: Eighty rats were divided into four groups to receive an intrathecal injection of normal saline containing either 5% lidocaine, 5% lidocaine with 0.2 mg/ml of epinephrine, 0.2 mg/ml of epinephrine, or normal saline alone. Animals were assessed for persistent sensory impairment using the tail-flick test administered 4 and 7 days after infusion. Animals were then killed, and the spinal cord and nerve roots were prepared for neuropathologic evaluation. RESULTS: Rats given 5% lidocaine developed persistent sensory impairment and histologic damage, and the addition of epinephrine resulted in a further significant increase in injury. Sensory function in animals given epinephrine without anesthetic was similar to baseline and did not differ from saline. Histologic changes in animals treated with epinephrine alone did not differ significantly from saline controls. CONCLUSIONS: The neurotoxicity of intrathecally administered lidocaine is increased by the addition of epinephrine. When making clinical recommendations for maximum safe intrathecal dose of this anesthetic, one may need to consider whether the solution contains epinephrine. PMID- 11388542 TI - The cannabinoid agonist WIN55,212-2 suppresses opioid-induced emesis in ferrets. AB - BACKGROUND: Cannabinoid receptor agonists reverse nausea and vomiting produced by chemotherapy and radiation therapy in animals and humans but have not been tested against opioid-induced emesis. This study tests the hypothesis that cannabinoid receptor agonists will prevent opioid-induced vomiting. METHODS: Twelve male ferrets were used. They weighed 1.2-1.6 kg at the beginning and 1.8-2.3 kg at the end of the experiments. All drugs were injected subcutaneously. WIN55,212-2, a mixed CB1-CB2 cannabinoid receptor agonist, was administered 25 min before morphine. Retches and vomits were counted at 5-min intervals for 30 min after morphine injection. RESULTS: Retching and vomiting responses increased with increasing morphine doses up to 1.0 mg/kg, above which the responses decreased. Previous administration of naloxone prevented morphine-induced retching and vomiting. WIN55,212-2 dose-dependently reduced retching and vomiting. The ED50 was 0.05 mg/kg for retches and 0.03 mg/kg for vomits. At 0.13 mg/kg, retching decreased by 76% and vomiting by 92%. AM251, a CB1 receptor-selective antagonist, blocked the antiemetic actions of WIN55,212-2, but AM630, a CB2 receptor selective antagonist, did not. CONCLUSIONS: These results demonstrate that WIN55,212-2 prevents opioid-induced vomiting and suggest that the antiemetic activity of WIN55,212-2 occurs at CB1 receptors. This is consistent with findings that CB1 receptors are the predominant cannabinoid receptors in the central nervous system and that antiemetic effects of cannabinoids appear to be centrally mediated. PMID- 11388543 TI - Current issues in spinal anesthesia. PMID- 11388544 TI - The role of World War II and the European theater of operations in the development of anesthesiology as a physician specialty in the USA. AB - World War II was a time of growth and development of anesthesia as a physician specialty. Wartime training exposed neophyte physician-anesthetists to role models who showed the potential of anesthesiology and to the richness of practicing anesthesia. Wartime anesthesia required dexterity, imagination, and pluck, and surgeons and other physicians were suitably impressed. Drawing historical conclusions about cause and effect is hazardous. Recognized and unrecognized biases, preconceived notions, and the quality and type of resources available affect writers. With this in mind, consider how the effects of World War II on the growth of physician anesthesia loosely parallel the growth of anesthesia in Great Britain during the 19th century. Anesthesia became a medical profession in Great Britain because of the interest and support of physicians and the complexity of administering chloroform anesthesia. Similarly, World War II physician-anesthetists showed they could provide complex anesthesia care, such as pentothal administration, regional anesthesia, and tracheal intubation, with aplomb and gained the support of surgical colleagues who facilitated their growth within a medical profession. They returned to a medium ready to support their growth and helped to establish the medical profession of anesthesiology in the United States. PMID- 11388545 TI - Relevant concentrations of inhaled anesthetics for in vitro studies of anesthetic mechanisms. PMID- 11388546 TI - Severe bronchospasm and desaturation in a child associated with rapacuronium. PMID- 11388547 TI - How serious is the bronchospasm induced by rapacuronium? PMID- 11388548 TI - Bronchospasm after rapacuronium in infants and children. PMID- 11388549 TI - High-frequency jet ventilation in life-threatening bilateral pulmonary contusion. PMID- 11388550 TI - Jet ventilation for fiberoptic bronchoscopy. PMID- 11388551 TI - Did anesthetics trigger cardiac arrests in patients with occult myopathies? PMID- 11388552 TI - Anesthesia-related cardiac arrest in children. PMID- 11388553 TI - Is difficult mask ventilation only correlated to the physical status of the patient? PMID- 11388554 TI - Mandibular protrusion test for prediction of difficult mask ventilation. PMID- 11388555 TI - Large hypopharyngeal tongue: a shared anatomic abnormality for difficult mask ventilation, difficult intubation, and obstructive sleep apnea? PMID- 11388556 TI - The costs of Medicare compliance. PMID- 11388558 TI - Head extension angle required for direct laryngoscopy with the McCoy laryngoscope blade. PMID- 11388557 TI - An intubating airway with teeth. PMID- 11388559 TI - Electrical short circuit as a possible cause of death in patients on PCA machines: report on an opiate overdose and a possible preventive remedy. PMID- 11388560 TI - Accidental extreme neck extension during repositioning of an operating room table. PMID- 11388561 TI - Assessing parenting competence in child protection cases: a clinical practice model. AB - Evaluating parents in the context of possible abuse or neglect involves unique challenges. This paper describes a practice model for conducting clinical evaluations of parents' ability to care for young children (under age 8). Core features of the model include (a) a focus on parenting qualities and the parent child relationship, (b) a functional approach emphasizing behaviors and skills in everyday performance, and (c) application of a minimal parenting standard. Several factors complicate the assessment task, namely, the absence of universally accepted standards of minimal parenting adequacy, the coercive context of the assessment, the scarcity of appropriate measures, difficulties predicting future behavior, and the likely use of the evaluation in legal proceedings. In the proposed model, the evaluator (a) clarifies specific referral questions in advance; (b) uses a multimethod, multisource, multisession approach; (c) organizes findings in terms of parent-child fit; (d) prepares an objective, behaviorally descriptive report that articulates the logic for the evaluator's clinical opinions regarding the referral questions; and (e) refrains from offering opinions regarding ultimate legal issues. The paper describes requisite skills needed to conduct parental fitness evaluations, sample methods, and a protocol for writing the evaluation report. PMID- 11388562 TI - Family processes in adolescent depression. AB - A developing body of research points to the role of family relationships and interactions as factors relevant to understanding unipolar depressive symptomatology and disorder in adolescents. This paper is focused on examining mechanisms by which adverse family processes may operate to increase adolescents' vulnerability to depression. The review is organized into a description and evaluation of four potential mechanisms by which families may influence adolescents' risk: (a) Stress/Support, (b) Social Interactional, (c) Cognitive, and (d) Affect Regulation. The presentation of the individual mechanisms is followed by a brief discussion of how they could be integrated to develop a fuller understanding of familial processes relevant to the etiology and maintenance of depressive conditions. Finally, we conclude with a discussion of overall limitations in the literature and recommendations for future research. PMID- 11388564 TI - The Child and Adolescent Functional Assessment Scale (CAFAS): review and current status. AB - Measures of impairment in psychological and behavioral functioning have a long history in the field of children's mental health, and appear particularly useful in eligibility determination, treatment planning, and outcome evaluation of services for children and adolescents with serious emotional disturbance (SED). One recently developed multidimensional measure of functional impairment-the Child and Adolescent Functional Assessment Scale (CAFAS; K. Hodges, 1989, 1997) has enjoyed widespread use nationwide. It has been adopted as a tool for making treatment eligibility decisions and documenting outcomes on a statewide level in more than 20 states and on a local level in dozens of research and demonstration projects. In this paper, the technical merits of the CAFAS are closely examined, with the conclusion that empirical evidence is lacking to support its valid use in making the types of treatment decisions for which it is currently being employed across the nation. Furthermore, there appears to be little concern among mental health researchers, practitioners, administrators, and state legislators about these apparent limitations of the CAFAS. The potential benefits of establishing objective and valid level-of-need criteria, using the CAFAS are numerous and the interest in doing so is clear; however, the psychometric limitations of the scale identified in this review need to be addressed before its full potential can be realized. PMID- 11388565 TI - The application of radionuclide antisense therapy for malignant tumours. PMID- 11388566 TI - Measurements of both hippocampal blood flow and hippocampal gray matter volume in the same individuals with Alzheimer's disease. AB - We evaluated both hippocampal blood flow and hippocampal gray matter volume using single photon emission tomography and magnetic resonance imaging in the same individuals with Alzheimer's disease (AD) and in age-matched controls. The hippocampal blood flow was not significantly lower in mild AD patients (n = 21, Mini-Mental State Examination (MMSE) 23.3+/-2.1) than in controls (n = 16) with a 57% overlap. The hippocampal blood flow was significantly lower in advanced AD patients (n = 22, MMSE 15.4+/-3.2) than in controls. The hippocampal gray matter volume was significantly smaller in mild AD patients than in controls, although a 43% overlap was present. There was no significant difference in the hippocampal gray matter volume between the mild and advanced AD patients. The combination of measurements of hippocampal blood flow and gray matter volume discriminated 71% of mild AD patients from controls. These results suggest the usefulness of a combined analysis of hippocampal blood flow and gray matter volume for the early diagnosis of AD. PMID- 11388563 TI - Social anxiety disorder in childhood and adolescence: current status and future directions. AB - This paper reviews the current status of research on the phenomenology, etiology, maintenance, assessment, and treatment of childhood and adolescent social anxiety disorder (SAD). Despite being one of the most prevalent disorders of childhood and adolescence, SAD paradoxically stands as one of the least recognized, researched, and treated pediatric disorders. The small treatment outcome literature provides preliminary support to the effectiveness of various forms of cognitive behavior therapy. The majority of studies to date, however, are limited by inadequate control conditions. Other findings include some support for the utility of parental involvement in treatment, significant advancements in outcome measures (e.g., normative comparisons, indices of naturalistic social functioning), and impressive durability of gains for the majority of treatments. Future directions are suggested, including experimental and naturalistic studies of developmental pathways and maintenance factors, the incorporation of "positive psychology" constructs (e.g., positive emotions, hope, self-control) in treatment and prevention, and the continued delineation of differences between child, adolescent, and adult manifestations of SAD. PMID- 11388567 TI - Increased uptake of 99Tcm-ethyl cysteinate dimer in patients with brain tumours. AB - We performed 229 99Tcm-ethyl cysteinate dimer (ECD) single photon emission tomography (SPET) studies in 185 patients with brain tumour. Increased uptake of the tracer was observed in 11 cases. In six of these 11 patients, focal intense activity was seen in the area surrounding the tumour. Five of these six patients had episodes of seizure, and ictal SPET showed further increased uptake in the area of hyperperfusion in one patient. Hyperperfusion surrounding the tumour might be related to seizure. In the remaining five patients, increased accumulation was seen in the tumour. Three of these five patients had a discrepancy between 99Tcm-ECD SPET and 201Tl SPET imaging. There could be some difference in the mechanism of accumulation in the tumour between 99Tcm-ECD and 201Tl. PMID- 11388568 TI - Positron emission tomography of the thyroid, with an emphasis on thyroid cancer. AB - The role of Positron Emission Tomography (PET) using 18F-fluorodeoxyglucose (FDG) in the management of thyroid cancer is discussed. It is important to ensure that patients are relaxed because uptake of FDG in tense or active muscles in the neck and larynx can be misinterpreted as metastases. The major role for PET is in patients where the stage of disease is uncertain, usually the result of discordant negative 131I scan and a positive serum thyroglobulin (Tg) values. PET identifies the source of Tg production in 50-80% of patients. PET scan can be negative in well differentiated cancers which retain the ability to trap iodine. This can result in a 'flip/flop', with negative PET, positive radio-iodine scan, or positive PET, negative radioiodine scan. PET is also valuable in identifying the source of calcitonin production in patients with medullary thyroid cancer. When focal uptake is seen in the thyroid of patients who are scanned for non thyroidal reasons, the likelihood of primary thyroid cancer is high. In contrast diffuse uptake of FDG in the thyroid is usually the result of auto-immune thyroid disorders. PMID- 11388569 TI - Adsorption of radiopharmaceuticals to syringes leads to lower administered activity than intended. AB - In this study we examined the delivered activity of radiopharmaceuticals injected into patients and the various factors which lead to incomplete delivery. Four commonly used 99Tcm radiopharmaceuticals were included, namely pertechnetate, methylene diphosphonate (MDP), macro-aggregated albumin (MAA) and tetrofosmin. There was no appreciable residual activity after injection of pertechnetate and MDP other than that due to the dead space in syringes (0.06 ml, 4% of measured activity). However, there was considerable residual activity following an MAA injection. This was particularly dependent on the radiopharmaceutical residence time in the syringe prior to injection. If MAA was pre-dispensed there was up to 80% of activity remaining in the syringe following injection. We have shown that this activity is not bound and can be decreased by agitating the syringe prior to the injection. For tetrofosmin, the activity remaining in the syringe was shown to be mostly due to adsorption of the tetrofosmin onto the plastic syringe (up to 30% of the activity); in particular, onto the rubber plunger. The amount of binding probably depends on the formulation of the rubber plunger and the plastic barrel, which varies between manufacturers. Of the three commonly used syringe brands (Becton-Dickinson, Sherwood, and Braun syringes) the Braun syringes were shown to have the least binding (6%) and would be most suitable for tetrofosmin injection. We recommend the use of Braun syringes for tetrofosmin. We also recommend that pre-dispensed MAA injections should be gently agitated before injection into the patient. No special precautions are necessary for pertechnetate or MDP injections. PMID- 11388570 TI - In vivo evaluation of three different 99mTc-labelled radiopharmaceuticals for sentinel lymph node identification. AB - This work was designed to compare sentinel lymph node (SLN) uptake of 99mTc labelled human serum albumin colloid (99mTc-HSAC), 99mTc-labelled antimony sulphur colloid (99mTc-SC) and a 99mTc-labelled dextran 70 solution (99mTc-Dx) and their selectivity in the identification of this node in the right rear footpad (RRF) of normal mice and tumour bearing mice. Radiopharmaceutical uptake in the SLN (popliteal lymph node) and the lumbar lymph node (LLN), the second lymphatic node station from RRF, were measured at different time points post intradermal or intratumoural injection into the RRF of NIH normal mice and of Balb/c mice harbouring the murine mammary tumour M2. 99mTc-HSAC uptake in the SLN was significantly higher than LLN uptake. The 99mTc-SC demonstrated high uptake in SLN, but accumulation in LLN was also high. 99mTc-Dx showed low uptakes in both SLN and LLN. The intradermal injection resulted in a more effective radiopharmaceutical accumulation in SLN than did the intratumoural inoculation. Data also show that increments in tumour volume reduced radiopharmaceutical uptake in the SLN. Our results show that 99mTc-HSAC exhibits the highest uptake in the SLN combined with the smallest amounts of radiopharmaceutical passing through to the LLN. Therefore, 99mTc-HSAC appears to be the best radiopharmaceutical for sentinel node detection. PMID- 11388571 TI - Quantification of intrahepatic non-uniform distributions for assessing impaired function of liver using 99Tcm-DTPA-galactosyl serum albumin liver SPECT scintigraphy. AB - Quantitative evaluation of intrahepatic non-uniform distribution was attempted by functional volumetric analysis of liver single photon emission computed tomography (SPECT) scintigraphy using 99Tcm-diethylenetriaminepentaacetic acid galactosyl serum albumin (99Tcm-GSA) (185 MBq). A total of 148 patients were classified into three groups: diffuse liver disease, liver disease with space occupying lesions, and non-hepatic disease. The volume obtained at the 10% cut off threshold from reconstructed liver-specific SPECT images was assumed to be the functional total hepatic volume, and the functional volume rates were calculated every 10% threshold width from 10% to 100%. The hepatic distribution index (D index) was calculated from the sum of absolute differences from the control. The hepatic D index in liver cirrhosis was significantly higher than in hepatitis and other diffuse liver diseases, and the index in liver disease with primary malignant neoplasm was significantly higher than in liver disease with metastasis or a benign-space occupying lesion. Additionally, it was significantly negatively correlated with LHL15, and positively correlated with the clinical severity. However, in stages II and III, with which LHL15 did not significantly correlate, the hepatic D index matched the severity of hepatic impairment. The hepatic D index evaluates distribution of functional hepatocytes and may provide a new clinical method for quantitatively assessing hepatic impairment. PMID- 11388572 TI - Imaging infection with LeuTech. AB - LeuTech is a 99Tcm labelled, anti-CD15, IgM, murine monoclonal antibody shown to have high affinity (Kd = 10(-11) M) for CD15 receptors (5.1 x 10(5)/cell) expressed on human neutrophils. LeuTech was injected directly, intravenously, and its efficacy in imaging infection in 46 consecutive patients was determined. Human anti-mouse antibody (HAMA) response was examined in 30 normal volunteers using a standard LeuTech dose reconstituted with decayed 99Tcm solution. There were 38 true positive, six true negative, and two false negative scans. Of the 38 positive images, 33 (92%) were positive within 10 min after injection of LeuTech. LeuTech accuracy in this group of patients was 96%, sensitivity 95%, specificity 100%, positive predictive value (PPV) 100%, and negative predictive value (NPV) 75%. No elevation of the HAMA titre was observed in any of the 30 normal volunteers and no adverse reaction was noted in any patient. LeuTech is a highly promising agent for rapid imaging of infectious foci. PMID- 11388573 TI - A physiological manoeuvre to improve the positivity of the gastro-oesophageal reflux scintigraphic test. AB - One of the best examinations used routinely to detect gastro-oesophageal reflux (GOR) is scintigraphy and attempts are continuously being made to improve the performance of the test. We have tested a new manoeuvre to improve the results. Three hundred and twenty eight patients (190 males and 138 females, aged 1 month to 84 years, average 4.47 years) were studied after a 12 h fast period with 100 MBq of 99Tcm-phytate, 50 ml volume, directly delivered into the stomach in a mixture of orange juice. All patients were strongly suspected of having GOR. Conventional acquisition was done for 20 min, at a rate of one frame every 20 s. In the middle of the examination, small children were put in the upright position and held there for a few seconds or released in a way that allowed movement; adults were asked to stand up and walk a few metres. The test was then continued for the remaining 10 min. The global index of positivity was 64.6% (76 positive cases in the first half of the examination and 134 positive cases only after the manoeuvre). The mean of GOR episodes observed in the positive cases without the manoeuvre was 0.5 per patient, significantly different from the mean of 1.59 noted after the manoeuvre (P<0.01). In 28 cases the manoeuvre increased either the intensity or the frequency of reflux previously detected without it. We believe that this manoeuvre should be performed in all scintigraphic tests aiming to detect GOR. In the present series, the manoeuvre increased the frequency of GOR episodes. PMID- 11388574 TI - Utility of ventilation and perfusion scan in the diagnosis of young military recruits with an incidental finding of hyperlucent lung. AB - Swyer-James-MacLeod syndrome (SJMS) is considered to be a relatively uncommon disease presenting with unilateral hyperlucent lung due to hypoplasia of a pulmonary artery and bronchiectasis of the affected lung. In this report, we describe the ventilation-perfusion (V/Q) scan findings of nine male recruits (aged 20-29 years, mean 24.4+/-2.96 years) with SJMS in whom the diagnosis was first established in adulthood. V/Q scan findings of all patients were compared with those on planar radiographs, pulmonary function studies, high resolution computed tomography (HRCT) and digital subtraction angiography (DSA). The ventilation (133Xe) and perfusion (99Tcm-macro-aggregated albumin) scans showed the characteristic pattern of a matched V/Q defect and marked air trapping on the washout phase on 133Xe scintigraphy. HRCT displayed hypodense lung with integrity of main airways, and markedly diminished vasculature on the affected side in all patients. A smaller pulmonary artery on the affected side with poor peripheral vasculature was observed with DSA in all patients. All patients had features of obstructive airway disease in varying degrees on pulmonary function studies. In contrast to other imaging methods, bronchiectasis as an etiological factor was displayed on HRCT. Some pulmonary areas, which were normal on HRCT and planar radiographs, showed air trapping on V/Q scan. Although a V/Q scan was more helpful in determining the extent of the disease and correlates well with conventional imaging methods, HRCT was the most valuable imaging method for the evaluation of aetiology in unilateral hyperlucent lung. PMID- 11388575 TI - Interpretive intra- and interobserver reproducibility of rest/stress 99Tcm sestamibi myocardial perfusion SPECT in a consecutive group of male patients with stable angina pectoris before and after percutaneous transluminal angioplasty. AB - BACKGROUND: Observer variability of 99Tcm-sestamibi myocardial perfusion imaging (MPI) has rarely been investigated. The aim of our study was to evaluate the interpretive reproducibility with this technique. PATIENTS: We report on 108 consecutive male patients with stable angina pectoris, investigated before and after percutaneous transluminal angioplasty (PTCA). METHODS: A 2-day rest/stress 99Tcm-sestamibi gated single photon emission computed tomography (SPECT) protocol was used. MPI was interpreted by two independent observers without knowledge of clinical data, using a 20-segment scoring model. RESULTS: Intra- and interobserver agreement was found to be good to excellent (kappa = 0.71-0.85) with regard to the overall diagnosis as well as the individual vessel diagnosis (kappa = 0.60-0.87). However, agreement was higher for left anterior descending coronary artery (LAD) and left circumflex coronary artery (LCX) vascular territories than for the right coronary artery (RCA) territory. Moderate to good intraobserver agreement (kappa = 00.54-0.68) and slightly lower interobserver agreement (kappa = 0.52-0.56) was found for segmental score interpretation. When comparing the interpretive reproducibility before and after PTCA intra- and interobserver agreement was better after PTCA, probably reflecting the increase in normal scans after revascularization. CONCLUSIONS: In a group of consecutive male patients with stable angina pectoris interpretive reproducibility (overall and individual vessel diagnosis) was good to excellent. However, segmental scoring reproducibility was moderate to good. PMID- 11388576 TI - Evaluation of histopathological differentiation in lung adenocarcinoma patients using 201Tl-chloride and 99Tcm-MIBI SPET. AB - The purpose of this prospective study was to evaluate the relationship between thallium-201 chloride (201Tl) and technetium-99m hexakis 2 methoxyisobutylisonitrile (99Tcm-MIBI) accumulation and histopathological differentiation in primary lung adenocarcinoma. A total of 43 patients with primary lung adenocarcinoma were investigated. The patients were divided into well differentiated (n = 17), moderately differentiated (n = 14) and poorly differentiated (n = 12) carcinoma groups. Simultaneous dual single photon emission tomography (SPET) images with 201Tl and 99Tcm-MIBI were acquired 15 min (early) and 2 h (delayed) after injection. Using a region of interest technique, the tumour-to-normal lung ratio was calculated for both early (early ratio) and delayed (delayed ratio) images. The retention index was calculated using the formula delayed ratio/early ratio. Uptake of the radionuclides was compared with the classification of tumour differentiation grading. The mean (+/- SD) values of the early ratio, delayed ratio and retention index using 201Tl were 2.19+/-0.72, 2.28+/-0.71 and 1.06+/-0.16, respectively, in the well differentiated group, 2.38+/-0.83, 2.48+/-0.84 and 1.08+/-0.23, respectively, in the moderately differentiated group, and 2.87+/-0.75, 3.60+/-1.51 and 1.22+/-0.21, respectively, in the poorly differentiated group. Both the ratios and the retention index using 201Tl were significantly lower in the well differentiated group than in the poorly differentiated group. The delayed ratio using 201Tl in the moderately differentiated group was also significantly lower than that in the poorly differentiated group. There were no significant differences in either ratio or the retention index among the three groups using 99Tcm-MIBI. 201Tl SPET is superior to 99Tcm-MIBI SPET for the grading of histopathological differentiation of primary lung adenocarcinoma. PMID- 11388577 TI - Is the depth correction using the geometric mean really necessary in a 99Tcm-DMSA scan in the paediatric population? AB - Determination of the left to right dimercaptosuccinic acid (DMSA) uptake ratio is theoretically one of the easiest quantitative procedures in nuclear medicine. The quantification can be performed on the posterior view, with or without the lateral view for correction of kidney depth. The geometric mean can also be determined using both the anterior and the posterior views. The aim of this study was to evaluate the occurrence of remarkable differences in the results from quantification of the relative renal function using the geometric mean and those obtained using the posterior counts only. Moreover, we evaluated to what extent the patient age influenced these differences. We reviewed 328 99Tcm-DMSA scans. The difference between the relative renal function obtained using the posterior view and that obtained using the geometric mean was calculated and analysed statistically. For the purpose of evaluating the value of performing the geometric mean calculation in patients of different ages, patients were divided into four age groups (group I, < or =2 years; group II, 3-9 years; group III, 10 18 years; group IV, >18 years). Using the Student's t test, no statistical differences were found in the relative renal function obtained by the two methods (posterior projection and geometric mean) in groups I (t = 0.01, P = 0.992) and II (t = 1.43, P = 0.155), which consisted of patients younger than 10 years (77% of the patients). In groups III and IV statistical differences were found (t = 2.27, P = 0.028 and t = 2.170, P = 0.038), respectively. We conclude that for children under 10 years it is unnecessary to perform depth correction using the geometric mean except in rare cases of major malformations and position anomalies. PMID- 11388578 TI - In vitro affinity of 99Tcm-labelled N2S2 conjugates of chrysamine G for amyloid deposits of systemic amyloidosis. AB - To date, systemic amyloidosis is diagnosed histologically in vitro using Congo red staining or in vivo using iodine-123 serum amyloid P component (123I-SAP) scintigraphy. 99Tcm-labelled derivatives of chrysamine G (CG), a lipophilic analogue of Congo red, were synthesized as potential tracer agents for direct and quantitative scintigraphic evaluation of amyloid deposits. To determine the affinity of 99Tcm-MAMA-CG, 99Tcm-Me4MAMA-CG and 99Tcm-MAMA-CG diethyl ester for amyloid, in vitro autoradiography was performed on sections of human kidney biopsy cylinders from kidneys with amyloid deposits (types AA, Alambda and Akappa) or control kidney tissue after incubation with the respective tracer agents. The binding of 99Tcm-MAMA-CG and its tetramethyl derivative was higher to kidney biopsy material with amyloid deposits of the AA, Alambda or Akappa type compared with control kidney tissue. This higher binding was prevented by the presence of 10 microM Congo red in the incubation medium. The diethyl ester of 9Tcm-MAMA-CG did not demonstrate increased binding to Congo red-positive kidney tissue. In conclusion, 99Tcm-MAMA-CG and 99Tcm-Me4MAMA-CG localize specifically to amyloid deposits in human kidney tissue, suggesting that these tracer agents may be applicable as specific targeting agents for diagnostic purposes in clinical amyloidosis. PMID- 11388579 TI - UK audit and analysis of quantitative parameters obtained from gamma camera renography. AB - The purpose of this study was to perform an audit of quantitative values obtained from gamma camera renography in the UK. Ten patient image sequences representing normal and pathological renal function were obtained from archived studies and distributed to hospitals in the UK. Hospitals were asked to measure five parameters: relative function, renogram time-to-peak (left and right), and whole kidney mean transit time (left and right). Details of methodology, software used and operator experience were requested. This allowed the influence of operational factors on variations in reported values to be examined. A total of 180 responses from 81 hospitals were received. Values reported for the parameters, together with other details supplied, were entered into Excel and SPSS for statistical analysis. Histograms representing the distribution of values were produced for each parameter. The largest variations were found for mean transit time and occasionally for time-to-peak. The effect of factors was assessed using nonparametric statistical tests applied independently to each renogram. For all the parameters, the hospital, UK region, supplier, computer and software version influenced variations in the reported values. Algorithm and site of background region were influencing factors for relative function, the background subtraction method influenced time-to-peak, and curve smoothing influenced mean transit time. PMID- 11388580 TI - National survey of imaging technologists in nuclear medicine 1998/99. AB - This survey investigated the nature of imaging staff and the distribution and structure of nuclear medicine departments in the UK. Total numbers of cameras and average per department were lower than in 1989/90 but higher than the 1992/93 survey. Total whole time equivalents, average per department and average per camera had increased. Excluding the Isle of Man, Wales had the lowest population per camera and Northern Ireland the highest. This survey identified 237 departments (90 independent, 116 integrated and 31 satellite), radiographers being the chief technologists in 64% and MTOs in 36%. Over half had one single headed camera but one third had dual- or triple-headed cameras. Chief technologist grades ranged from Basic to Superintendent II and MTO3 to MTO5. The most common grades for other staff were Senior II or MTO3. Of the 786 technologists who provided details, 68% had initially trained as radiographers. Specific nuclear medicine qualifications were held by 67% of all technologists. In 52% of departments at least one member of staff rotated between nuclear medicine and another department or hospitals and 25% had no full-time staff. Survey returns revealed that, although 84% of imaging technologists often or occasionally attended conferences, 5% never attended. PMID- 11388582 TI - Improved sentinel node visualization in breast cancer by optimizing the colloid particle concentration and tracer dosage. AB - Faint lymph uptake may hamper sentinel node (SN) identification by scintigraphy and subsequent gamma probe localization. The aim of the present study was to evaluate an adjustment in the colloid particle concentration and tracer dosage to optimize mammary lymphoscintigraphy. Scintigraphy was performed in 151 patients with a palpable breast carcinoma and clinically negative axilla: for the first 75 patients (group A) a standard labelling of 0.5 mg nanocolloid with 99Tcm was performed, for the subsequent 76 patients (group B) the labelling dilution volume was reduced from 4 to 2 ml. For both groups the volume of injection was 0.2 ml. Lymph node uptake was evaluated by a 4-step visual score (from 0 = absent to 3+ = very intense), and by count quantification of at 4 h in the first draining SN. The SN visualization rate increased from 93% (70/75) in group A (mean dosage 93.4 MBq, range 57-130 MBq) to 99% (75/76) in group B (mean dosage 106.5 MBq, range 74 139 MBq). The percentage of patients with uptake 3+ was significantly higher (P = 0.001) in group B (51% vs 35% in group A). SN counts were significantly higher for group B (P<0.001). The percentage of patients with less than 2000 counts/node diminished from 45% in group A to 9% in group B (P = 0.001). In group B (P = 0.033) more lymph channels (53% vs 35% in group A) were visualized and for a longer time (26% vs 4% at 4 h). Axillary drainage was seen in 96% in group A and 98% in group B whereas non-axillary drainage was observed in 19% and 25%, respectively. Intraoperative SN identification rate was 97% in group A and 100% in group B. SN metastases were found in 41% of group A and 47% of group B. It is concluded that enhancement of colloid particle concentration and adjustment of tracer dosage led to improved SN identification by substantial increase in lymph node uptake and lymph vessel depiction. A significant reduction of cases with faint SN uptake enables better surgical efficacy. PMID- 11388581 TI - Statin induced myopathy does not show up in MIBI scintigraphy. AB - Statin induced myopathy is the most commonly seen side effect in users of this family of drugs. Their different forms present with either creatine phosphokinase (CK) elevation or not, signs of in vivo oxidation injury or not or a combination of both. The pathogenetic background, however, still remains obscure. As MIBI, beside myocardial and tumour scintigraphy, is useful in detecting muscle metabolic abnormalities, an increased uptake of MIBI in the diseased muscular segments could be expected. We investigated seven patients (five males, two females; aged 36-56 years) with statin induced myopathy with either elevated CK, isoprostanes or muscle pains at varying combinations. MIBI whole-body imaging was done immediately, the patients still being on the respective statin. Sixteen patients (six males, 10 females) suffering from lung or breast cancer and being on statins served as controls. No uptake abnormalities in any muscular segment either in the patients or the control group were seen. Thus, MIBI scintigraphy is not useful, apparently, in diagnosing and eventually localizing statin induced myopathy. These findings indicate that MIBI scintigraphy is of no help for diagnosis and gaining further insight into statin induced myopathy. PMID- 11388583 TI - Rapid imaging of human melanoma xenografts using an scFv fragment of the human monoclonal antibody H11 labelled with 111In. AB - H11 is a human IgM monoclonal antibody which recognizes a novel tumour-associated antigen expressed on melanoma, glioma, breast cancer, colon cancer, prostate cancer, lung cancer and B-cell lymphoma. In this study, a recombinant single chain Fv (scFv) fragment of H11 labelled with 111In was investigated for tumour imaging in athymic mice implanted subcutaneously with A-375 human melanoma xenografts. H11 scFv was derivatized with diethylenetriaminepentaacetic acid (DTPA) for labelling with 111In. The immunoreactivity of DTPA-H11 scFv against A 375 cells in vitro ranged from 23% to 36%. 111In-DTPA-H11 scFv was rapidly eliminated from the blood and most normal tissues (except the kidneys) reaching maximum tumour/blood ratios of 12:1 at 48 h post-injection. Tumours were imaged as early as 40 min after injection. The kidneys accumulated the highest concentration of radioactivity (up to 185% injected dose/g). Tumour uptake was 1 3% injected dose/g. The whole-body radiation absorbed dose predicted for administration of 185 MBq of 111In-DTPA-H11 scFv to humans was 37 mSv. The radiation absorbed dose estimates for the kidneys, spleen and intestines were 405 mSv, 698 mSv and 412 mSv, respectively. The results of this preclinical study and a concurrent phase I trial suggest a promising role for H11 scFv for tumour imaging. PMID- 11388585 TI - Biliary scintigraphy in the investigation of acalculous gallbladder disease. PMID- 11388584 TI - 18F-FDG labelling of human leukocytes. PMID- 11388586 TI - Detection of insomnia in primary care. AB - Insomnia is a widespread condition with diverse presentations. Detection and diagnosis of insomnia present a particular challenge to the primary care physician. Patients seldom identify their sleep habits as the source of the complaints for which they are seeking treatment. Insomnia may be the result of many different medical or psychiatric illnesses or the side effects of medications or legal or illegal recreational drugs. Insomnia has a serious impact on daily activities and can cause serious or fatal injuries. With ever-increasing competition with sleep from 24-hour television broadcasts from hundreds of channels and the Internet, as well as more traditional distractions of late-night movies, clubs, and bars, we have become a society that sleeps 25% less than our ancestors did a century ago. We have no evidence, however, that we require less sleep than they did. This article presents strategies for detecting and diagnosing insomnia. PMID- 11388588 TI - New developments for treating sleep disorders. PMID- 11388587 TI - A psychiatric perspective on insomnia. AB - Insomnia is a cardinal symptom for many psychiatric disorders, especially depressive disorders. Treatment of the underlying psychiatric disorder may be sufficient to relieve the accompanying insomnia. If the insomnia fails to respond, then consideration should be given to the possibility of inadequate treatment of the primary psychiatric disorder, iatrogenic insomnia, insomnia related to a medical disorder, or learned/habit insomnia. Persistent insomnia should be aggressively pursued, since it has been associated with a variety of adverse outcomes in samples of depressed patients. The physician should always inquire about and encourage healthy sleeping behaviors, even if hypnotic medication is contemplated. Benzodiazepines and nonbenzodiazepine benzodiazepine receptor agonists (BzRAs) have the best evidence for efficacy as hypnotics, although sedating antidepressants are popularly prescribed. Although all benzodiazepine hypnotics and nonbenzodiazepine BzRAs are comparably efficacious in inducing sleep, they vary markedly in their potential for residual side effects. PMID- 11388589 TI - Consequences of insomnia and its therapies. AB - Insomnia has a number of clinically significant associations. In particular, patients with chronic insomnia have higher rates of psychiatric and medical illnesses, and insomnia is an important risk factor in the development of depression. Insomniacs also have higher rates of health care utilization and reduced quality of life. This article reviews important clinical correlates of insomnia and evidence for possible causal relationships between sleep and health. PMID- 11388590 TI - Future directions in the management of insomnia. AB - Research on insomnia has provided a number of important new insights, but fundamental deficits in our understanding remain. In considering priorities for future research, 3 areas warrant immediate attention. First, a causal relationship between insomnia and the adverse outcomes seen in insomnia patients needs to be established. Second, currently available symptomatic therapies need to be optimized. Recent data suggest that some benzodiazepine receptor agonists produce their hypnotic effect without side effects that were presumed to be inherent to sedation. Understanding the neuropharmacology underlying this differential effect would allow substantial improvements in the risk-benefit ratio for these drugs. Finally, the mechanisms of insomnia need to be better understood. Several lines of evidence suggest that physiologic arousal is important to the clinical presentation of primary insomnia. It remains unclear, however, whether this activation is primary or secondary to the insomnia itself. If physiologic hyperarousal causes primary insomnia, it would provide new approaches to the management of this disorder. PMID- 11388592 TI - Sleep architecture and its relationship to insomnia. AB - The methods used to obtain and depict sleep data shape our understanding of sleep as a phenomenon. The standard criteria for describing sleep were developed in the late 1960s. These criteria, which were established on the basis of the polysomnographic equipment available at that time, called for the division of sleep into stages according to depth; the visual depiction of these stages led to the now widely accepted concept of "sleep architecture." Although the sleep architecture model remains useful, the technology that provided the model's framework for understanding sleep has been superseded by computer-assisted systems for recording and analyzing sleep that may allow us to acquire data on sleep that were unobtainable with older equipment. Future gathering and depiction of sleep data, regardless of the recording and assessment methods used, should minimize disruption of sleep during study, allow for computerized analysis of sleep parameters, and describe the data from the perspective of the effect that sleep and the problems surrounding it have on daytime functioning. PMID- 11388591 TI - Neurotransmitters and sleep. AB - Sleep is an active process, not just a default state when there is less incoming sensory information. It can be understood best by considering fluctuating levels of a series of neurotransmitters including the biogenic amines and acetylcholine. The effects of these neurotransmitters are not unique to sleep, but also subserve a wide range of other functions, including affect, sexual behavior, and appetite. The mechanism by which the most common hypnotics work is by binding to the benzodiazepine recognition site of the gamma-aminobutyric acidA-benzodiazepine receptor complex, which mediates action of the most widely distributed inhibitory neurotransmitter in the nervous system. It is possible that some endogenous sleep factors indirectly alter the properties of this receptor complex. PMID- 11388593 TI - Molecular analysis and long-term follow-up of patients with different forms of 6 pyruvoyl-tetrahydropterin synthase deficiency. AB - The outcome of 6-pyruvoyl-tetrahydropterin synthase (PTPS) deficiency, the most common form of tetrahydrobiopterin (BH4) deficiency, depends on factors such as severity of the disease, type of mutation, time of diagnosis, and mode of treatment. We investigated five patients from four different families, four of them presenting with the severe form of PTPS deficiency and one with the mild peripheral form. In this study, missense (L26F, T67M, P87L, V124L, D136G, D136V) and nonsense (R15-16ins) mutations were detected by reverse transcriptase polymerase chain reaction and sequence analysis. Two patients with the severe form were compound heterozygotes (T67M/P87L and D136G/R15-16ins), two siblings were homozygous for the D136V mutation, and in the patient with the mild form, heterozygous L26F/V124L mutations were present. Two patients are on combined therapy with L-dopa/carbidopa/5-hydroxytryptophan plus BH4, the siblings are on monotherapy with BH4, and the patient with the mild form is now off treatment, presenting with normal plasma phenylalanine levels. CONCLUSION: Long-term follow up shows that the outcome of 6-pyruvoyl-tetrahydropterin synthase deficiency benefits from treatment started in the first months of life and that the phenotype may change with age. Additionally, depending on the type of mutations, prenatal damage to the fetus may multiply the clinical abnormalities and thus worsen the prognosis of the disease. In patients initially diagnosed with the mild peripheral form of the disease, therapy with tetrahydrobiopterin should be stopped after some time to test whether hyperphenylalaninaemia was only a transient condition. PMID- 11388594 TI - Clinical and neuropsychological outcome in 33 patients with biotinidase deficiency ascertained by nationwide newborn screening and family studies in Austria. AB - Newborn screening for biotinidase deficiency (BD) provides prevention of neurological sequelae in patients with low residual enzyme activity by early treatment with oral biotin substitution. Screening 1.1 million newborns in Austria and consecutive family studies led to the identification of 21 patients with profound BD (residual activity <10%) (incidence: 1:59,800) and to 12 patients with partial BD (residual activity 10%-30%) (incidence 1:89,700). Application of an HPLC assay using the natural substrate biocytin allowed exact quantification of extremely low residual biotinidase activities and thus subdivision of patients with profound BD into a group with a residual activity 0% 1% of normal activity (n = 5) and >1%-<10% (n = 16) respectively. Evaluation of clinical and neuropsychological outcome showed that only patients with a biotinidase activity < 1% (n = 3/5) exhibited characteristic clinical symptoms within the first weeks of life, while five patients with a residual activity of 1.2%-4.6% did not develop clinical symptoms even when not treated until 3.5 21 years. In all patients with residual activity <10% and biotin substitution within the first weeks of life, neuropsychological outcome was normal, while abnormal in three out of five patients tested for IQ and treated after the age of 3.5 years. In five out of nine patients with poor compliance or delayed or no treatment, visual and brainstem auditory evoked potentials were measured and were within age related normal values. All patients with partial BD available for follow-up remained clinically and neuropsychologically asymptomatic without treatment at ages 2.5 10 years. CONCLUSION: The incidence of biotinidase deficiency in Austria is comparable to other European countries. Subdivision of the group of patients with profound biotinidase deficiency suggests that only patients with residual activities < 1% are prone to develop clinical symptoms early in life, while patients with residual activities >1% may remain asymptomatic even without treatment, as do patients with partial deficiency. Moderate mental retardation might represent a possible manifestation of cerebral dysfunction in patients with profound biotinidase deficiency. PMID- 11388595 TI - Genetic analysis of carbamoylphosphate synthetase I and ornithine transcarbamylase deficiency using fibroblasts. AB - Deficiencies of carbamoylphosphate synthetase or of ornithine transcarbamylase, two urea cycle enzymes located within mitochondria, often present as severe neonatal hyperammonaemic crises and have a poor prognosis. While genetic analysis of the X-chromosomal transmitted ornithine transcarbamylase deficiency (OTC) is performed by exon-wise mutation screening of genomic DNA in most cases, identification of mutations in the autosomal inherited carbamoylphosphate synthetase (CPS 1) deficiency requires analysis of transcripts due to the unknown genomic structure. We tested the hypothesis that CPS 1 and OTC are expressed at low levels in fibroblasts and indeed were able to amplify full-length cDNA from that source. Using a reverse transcriptase polymerase chain reaction based procedure we completely characterised the genetic background in five patients and identified three novel mutations and a novel polymorphism of the CPS 1 gene (deletion/insertion 2170delGCTCinsCCA, nonsense mutation 2359C > T, missense mutation 3161T > G and Thr1406Asn, respectively), as well as the missense mutations 482A > G and 994T > A of the OTC gene. CONCLUSION: Cultured fibroblasts are an easily accessible source for genetic analysis of inborn errors of urea cycle enzymes which are functionally expressed only in liver and gut. PMID- 11388596 TI - Latent coeliac disease in a child with epilepsy, cerebral calcifications, drug induced systemic lupus erythematosus and intestinal folic acid malabsorption associated with impairment of folic acid transport across the blood-brain barrier. AB - A 15-year-old boy with epilepsy and cerebral calcifications, treated with valproic acid, ethyl phenylbarbiturate and ethosuximide, was referred for drug induced systemic lupus erythematosus. Anti-gliadin (AGA) and anti-endomysium (EMA) antibody tests were both positive (EMA titre 1:50). Endoscopic duodenal biopsy showed intense chronic inflammation without villous atrophy or crypt hyperplasia. The child was discharged with a gluten-containing diet. The follow up showed an increase in EMA titre (1:200) and the persistence of AGA. After 15 months, a second endoscopic intestinal biopsy showed flat mucosa and villous atrophy. Three serum folic acid determinations showed 1.8, 2.4, 2.0 ng/ml (reference range 2.5-16.9 ng/ml) prior to the two intestinal biopsies, but returned to normal levels (11.8 ng/ml) after a gluten-free diet and oral supplementation together. Two years later, the frequency of epileptic seizures was unchanged despite ongoing anti-epileptic treatment and a gluten-free diet. As cerebral calcification and epilepsy are reminiscent of the findings in congenital folate malabsorption, oral loading tests with 5 mg folic acid were carried out and showed impaired intestinal absorption and a defect in the transport across the blood-brain barrier. Low CSF folate levels (13.9 and 12.6 ng/ml, reference range 15-40 ng/ml) and an alteration in the CSF/serum folate ratio (1.43 and 1.16, normal ratio 3:1) were also found as well as increased levels of cystathionine both in CSF (40 micromol/l, reference range 18-28 micromol/l) and in serum (32 micromol/l, reference value <0.10 micromol/l). CONCLUSION: Impairment of intestinal folic acid absorption with a defect in folic acid transport across the blood-brain barrier has been demonstrated in a case of epilepsy and cerebral calcifications associated with coeliac disease. PMID- 11388597 TI - Severe reversible renal failure due to naproxen-associated acute interstitial nephritis. AB - Acute interstitial nephritis is uncommon in children and has very rarely been described with naproxen treatment. We report the occurrence of severe acute renal failure in a 10-year-old girl with juvenile rheumatoid arthritis after 1 month of naproxen therapy. Renal biopsy showed severe acute interstitial nephritis. The patient recovered completely after discontinuation of naproxen and administration of methylprednisolone. A review of the literature regarding non-steroidal anti inflammatory drug-associated acute interstitial nephritis is provided. CONCLUSION: In an era of increasing popularity of non-steroidal anti-inflammatory drugs for use in children, paediatricians should be aware of the potential renal complications of this class of drugs. PMID- 11388598 TI - Fatal outcome in a female monozygotic twin with X-linked hypohydrotic ectodermal dysplasia (XLHED) due to a de novo t(X;9) translocation with probable disruption of the EDA gene. AB - Ectodermal dysplasias are a group of congenital disorders with defective development of the epidermis and its appendages. X-linked hypohydrotic ectodermal dysplasia (XLHED; OMIM 305100) is the most common form of ectodermal dysplasia. We report on two monozygotic twin girls with XLHED due to a t(X;9) translocation causing a disruption of the EDA gene and non random inactivation of the normal X chromosome. One of the girls died unexpectedly at 2.5 years of age. Autopsy revealed that lack of normal tracheobronchial secretions leading to complete tracheal obstruction by mucous debris was the probable cause of death. CONCLUSION: Morbidity and mortality of ectodermal dysplasias in infancy and early childhood can be significant. Early diagnosis by paediatricians is important and complications should be anticipated. PMID- 11388599 TI - Combined body plethysmographic, spirometric and flow volume reference values for male and female children aged 6 to 16 years obtained from "hospital normals". AB - Vital capacity (VC) and its subdivisions (IC and ERV), total lung capacity (TLC), residual volume (RV), peak expiratory flow (PEF), forced vital capacity (FVC), forced expiratory volume in one second (FEV1), maximum flow volume curve (MEF75, MEF50, MEF25, MMEF, FEF75-85), airway resistance (Rtot, Reff) and the thoracic gas volume at resting expiratory position (FRC) were measured in 187 girls and 213 boys (hospital normals) aged 6 to 16 years. The measurements were carried out consecutively on the same subjects in the morning using a volume-constant plethysmograph (Master-Lab, E. Jaeger; programs: body plethysmography, spirometry and flow volume). Using multiple regression analysis the best fitting curves for the prediction of normal values for boys and girls were selected. Analyses of covariance were performed to compare the adjusted means of the spirometric and body plethysmographic variables of the male and female subjects. As expected, we found higher static and dynamic (FVC, FEV1, PEF) lung volumes in boys than in girls relating to height. The flows (MMEF, MEF50, MEF25, FEF75-85) were significantly lower in the male than in the female subjects of the same age justifying separate prediction equations, but the same equation for both genders may be used for the resistance variables Rtot and Reff. Our results are compared with those of previous studies. CONCLUSION: Lung volumes and flows differ significantly between girls and boys calling for separate reference values for female and male subjects of the same age. PMID- 11388600 TI - Two hyperandrogenic adolescent girls with congenital portosystemic shunt. AB - We describe two adolescent girls with a congenital portosystemic shunt who exhibited hyperandrogenism in addition to insulin resistant hyperinsulinaemia. Case 1 was referred to our clinic to undergo a routine clinical work-up prior to tonsillectomy at 14 years of age. Mild liver dysfunction was identified and hypogenesis of the portal vein with a congenital portosystemic shunt diagnosed. Primary amenorrhoea and virilization were evident and an endocrinological evaluation revealed hyperandrogenism and insulin resistant hyperinsulinaemia. Case 2 was referred at 15 years of age because of cardiomegaly. Mild liver dysfunction and hyperbilirubinaemia led to a diagnosis of agenesis of the portal vein with a congenital portosystemic shunt. Virilization was evident and an endocrinological evaluation revealed hyperandrogenism and insulin resistant hyperinsulinaemia. The haemodynamics of these patients were similar to those of secondary portosystemic shunt due to liver cirrhosis, which is often associated with hyperinsulinaemia and/or non-insulin dependent diabetes mellitus. On the other hand, hyperandrogenism is associated with certain insulin-resistant conditions with hyperinsulinaemia, including the polycystic ovary syndrome (PCO). Hyperinsulinaemia is believed to cause hyperandrogenism in patients with PCO by stimulating androgen production in both the ovary and adrenal gland. Therefore, in congenital portosystemic shunts, hyperinsulinaemia is also thought to cause hyperandrogenism due to the same mechanism. CONCLUSION: A certain percentage of female patients with hyperandrogenism, likely including those with polycystic ovary syndrome may also have congenital portosystemic shunts. Our results indicate that serum levels of total bile acids and ammonia are prognostic indicators of this hepatic vascular anomaly. PMID- 11388601 TI - Where are all the babies? Paediatric risk of mortality score in meningococcal disease. PMID- 11388602 TI - Successful treatment of phenylketonuria with tetrahydrobiopterin. PMID- 11388604 TI - Reference data for 4- and 5-year-old-children on the Balance Master: values and clinical feasibility. AB - Reference data for children using the Balance Master may be of clinical interest and use in the management of postural control in a wide range of conditions. PMID- 11388603 TI - Bartonella henselae bacteraemia in patients with cat scratch disease. PMID- 11388605 TI - Neonatal screening for medium chain acyl-CoA deficiency: high incidence in Lower Saxony (northern Germany). PMID- 11388606 TI - Evaluation of weight gain composition during the first 2 months of life in breast and formula-fed term infants using dual energy X-ray absorptiometry. AB - Healthy term breast- and adapted formula-fed infants show similar weight gain and weight gain composition during the first months of life. PMID- 11388607 TI - Transient renal tubular acidosis in a neonate following transplacental acetazolamide. AB - Renal tubular acidosis (RTA) was observed in a preterm boy shortly after birth. His mother had glaucoma and had been treated during pregnancy with oral acetazolamide, a carbonic anhydrase inhibitor. When RTA developed, acetazolamide was detected in his serum demonstrating transplacental acetazolamide passage. PMID- 11388608 TI - Rodent malarias: the mouse as a model for understanding immune responses and pathology induced by the erythrocytic stages of the parasite. PMID- 11388609 TI - The intercellular adhesin locus ica is present in clinical isolates of Staphylococcus aureus from bacteremic patients with infected and uninfected prosthetic joints. AB - Although polysaccharide intercellular adhesin (PIA) is thought to be crucial in the pathogenesis of prosthetic device infections caused by Staphylococcus epidermidis, its role in prosthetic device infections caused by Staphylococcus aureus is unknown. To assess the clinical impact of PIA production, isolates from 15 prospectively identified cases of S. aureus bacteremia in patients with prosthetic joints (8 infected, 7 uninfected) were characterized for biofilm production, hemagglutination, and the presence of a 419-bp amplification product within icaA. Although icaA was present in all 15 isolates, none of the isolates produced hemagglutination and only one isolate (from a patient with an uninfected prosthetic device) weakly produced biofilm in vitro. These results support the observation that the ica locus is conserved between S. epidermidis and S. aureus and that PIA may be expressed only under in vivo conditions. Future investigations should include animal models to approximate the complex milieu surrounding implanted prosthetic medical devices. PMID- 11388610 TI - The detergent octylglucoside neutralizes lipopolysaccharide in a specific manner. AB - The stimulatory effect of lipopolysaccharide (LPS) on human macrophages was found to be neutralized by the detergent octylglucoside (OG). Both macrophage stimulation and reactivity in a limulus amebocyte lysate test were suppressed by suspension of LPS in OG at concentrations between 0.25 and 2.5 mM, whereas other stimulatory lipopeptides and lipid containing stimulants were unaffected by OG. LPS at concentrations causing maximal stimulation of macrophages could be completely neutralized by non-toxic concentrations of OG. In addition, it was found that the neutralization in complex mixtures of macromolecules, such as bacterial cell lysate, was specific for LPS and that the stimulatory activity of the other substances in the mixture was not affected by the OG. PMID- 11388611 TI - Successful in vitro generation of Epstein-Barr virus-specific cytotoxic T lymphocytes from severe chronic active EBV patients. AB - Severe chronic active Epstein-Barr virus (EBV) infection (SCAEBV) is a rare but life-threatening disorder. Poor cytotoxic activity against the virus is widely believed to contribute to the development of this disease. We wished to determine whether it is possible to generate autologous EBV-specific cytotoxic T cells (CTLs) in vitro that can be infused back into the patient to treat his/her viremia. To do this, we first had to establish autologous EBV-transformed B cells (EBCL) as antigen-presenting cells, which is known to be difficult to do with B cells from SCAEBV patients. In one patient, the standard method of incubating B cells with EBV-containing B95-8 supernatant was sufficient. In a second patient, however, the B cells apoptosed too rapidly in culture to permit transformation. However, apoptosis could be blocked by the presence of CD40 ligand-transfectant cells, and EBV transformation was successful when performed with this transfectant. Indicating a native immune response to EBV, peripheral blood lymphocytes from both patients proliferated in response to autologous EBCL. Furthermore, patient T cells had higher frequencies of IFN-gamma-producing CD8 cells after stimulation with autologous EBCL than sero-positive healthy controls. EBV-specific CTLs could be generated from both patients after repeated stimulation with autologous EBCL. These CTL lines were predominantly composed of CD4+ cells, and autologous EBCL killing was largely inhibited by an antibody against HLA-DR. These findings support the possibility of adoptive immune therapy to treat SCAEBV patients. PMID- 11388612 TI - A dietary probiotic (Bifidobacterium lactis HN019) reduces the severity of Escherichia coli O157:H7 infection in mice. AB - The protective effects of the probiotic Bifidobacterium lactis HN019 against Escherichia coli O157:H7 were investigated in murine challenge infection models. BALB/c or C57BL/6 mice were fed milk-based diets supplemented with B. lactis HN019 (3 x 10(8) cfu/g) for 7 days prior to and following oral challenge with E. coli O157:H7. Behavioral parameters (morbidity, feed intake) were measured for 7 days following challenge; immunological responses (phagocytosis, antibody) and pathogen translocation were measured in a sub-sample of ostensibly healthy animals 1 week post-challenge. Results showed that HN019-fed mice maintained significantly higher post-challenge feed intake and exhibited a lower cumulative morbidity rate, compared to control mice which did not receive the probiotic. Significantly higher proportions of phagocytically active cells in the blood and peritoneum, and higher intestinal tract IgA anti-E. coli antibody responses, were recorded among HN019-fed mice compared to controls. Among HN019-fed mice, pathogen translocation was identified in one of five BALB/c and one of five C57 mice; the comparative figures in control mice were two of five and three of five, respectively, and the mean bacterial burdens in these mice were over 100-fold higher than in HN019-fed mice. These results demonstrate that HN019 can reduce the severity of infection due to the enterohemolytic pathogen E. coli O157:H7, and suggest that this reduction may be associated with enhanced immune protection conferred by the probiotic. PMID- 11388613 TI - Two different NO-dependent mechanisms account for the low virulence of a non mycelial morphological mutant of Candida albicans. AB - We have previously described the low virulence of a Candida albicans morphological mutant: 92'. We have now used this strain to examine the role of phagocytes in its pathogenesis. Our results show that C. albicans 92' cannot evade innate host macrophage defence mechanisms as efficiently as the parental strain. In addition to the high susceptibility to phagocytosis by peritoneal macrophages, the NO produced by macrophages is a very important element in the low virulence of this agerminative mutant, a thesis supported by in vivo and in vitro experiments. Whereas the parental strain was able to inhibit macrophage NO production, the mutant was quite inefficient at reducing NO production by macrophages. In addition, the mutant showed high sensitivity to a NO generator. Treatment of mice with aminoguanidine (a preferred inducible NO synthase inhibitor) caused 90% mortality in 92' systemic infection, thus supporting a role for NO in the low virulence of this strain. Our data show that both the low inhibitory effect of 92' on macrophage NO production and the higher sensitivity to NO underlie the low virulence of this strain. PMID- 11388614 TI - Neopterin production and tryptophan degradation in humans infected by Streptococcus pyogenes. AB - Streptococcus pyogenes may cause tonsillitis, scarlet fever and so-called "streptococcal toxic shock-like syndrome" (STSS). These streptococci produce exotoxins which are implicated as superantigens in the pathogenesis of STSS and scarlet fever. Using human peripheral blood-derived mononuclear cells in vitro, such toxins were shown to induce neopterin production and degradation of the amino acid tryptophan to metabolites such as kynurenine by activating indoleamine (2,3)-dioxygenase via interferon-gamma. We investigated the sera of seven patients with streptococcal tonsillitis and of four patients with STSS. Those with STSS showed higher serum neopterin concentrations (median: 152 nmol/l; 95th percentile in healthy controls: 8.7 nmol/l) than those with tonsillitis (median: 12 nmol/l). Similarly, kynurenine to tryptophan ratios were increased in tonsillitis and extremely high in patients with STSS. Highly increased neopterin production and tryptophan degradation in patients with STSS suggest an association between a high degree of T cell activation and the severity of the disease manifestation. PMID- 11388615 TI - Absence of anti-hepatitis B surface antibody after vaccination does not necessarily mean absence of immune response. AB - A small number of subjects vaccinated against hepatitis B do not produce anti hepatitis B surface (HBs) antibody levels detectable by commercial assays. Others lose detectable anti-HBs at some time after vaccination. The absence of clinical hepatitis despite potential exposure to hepatitis B virus (HBV) in both kinds of subjects suggests that they might be protected by low antibody levels. However, besides anti-HBs, T helper response and memory cells which may be induced by the vaccine are certainly also important for immunity against HBV. In the present study, samples from vaccinated subjects, found to be anti-HBs negative in an initial assay, subsequently showed positive results in, respectively, 25%, 36% and 38% of the cases, when a second, third and fourth assay was used. In addition, 360 samples from "nonresponders" and from vaccinees who had lost anti HBs, the reactivity of which was under the enzyme-linked immunoassay-cut-off value were compared to that of nonvaccinated controls. The absorbances were found to be significantly higher in the nonresponders (0.038) and in the vaccinees having lost anti-HBs (0.041), than in the controls (0.025). Such findings contribute to explaining why so-called nonresponders as well as vaccinees who have lost anti-HBs nevertheless appear to be protected. PMID- 11388616 TI - Cecropins, antibacterial peptides from insects and mammals, are potently fungicidal against Candida albicans. AB - Natural products are the major source of lead compounds for drugs against human pathogens. Among the first natural peptides from animals for which a potent antibacterial activity has been recognized were the cecropins. The 30- to 40 residue alpha-helical peptides display their activity by permeabilizing the membranes of bacteria. Although originally isolated from insect hemolymph, a structural and functional correlate was also found in a mammal. Here, we report on the finding that cecropin A and B from the silk moth Cecropia as well as the porcine cecropin P1 are capable of inhibiting the growth of and to kill yeast phase Candida albicans. The peptides were tested in radial diffusion and microbroth dilution assays. They displayed potent activity against a clinical isolate as well as against defined culture strains of the pathogenic yeast but are of exceedingly low cytotoxicity towards the human cell line Jurkat. The candidacidal properties of the intensely studied molecules known to be highly active against Gram-positive and Gram-negative bacteria may renew the interest in these natural broad-spectrum peptide antibiotics and their limited cytotoxicity to human cells may be exploited for the development of topical therapeutics against pathogens resistant to classical antibiotics. PMID- 11388617 TI - Doppler echocardiographic assessment of subclinical valvitis in the diagnosis of acute rheumatic fever. PMID- 11388618 TI - Can subclinical valvitis detected by echocardiography be accepted as evidence of carditis in the diagnosis of acute rheumatic fever? AB - AIM: Subclinical valvar insufficiency, or valvitis, has recently been identified using Doppler echocardiography in cases of acute rheumatic fever with isolated arthritis or chorea. The prognosis of such patients with acute rheumatic fever and subclinical valvitis is critical when determining the duration of antibiotic prophylaxis. We aimed, therefore, prospectively to investigate the association of silent valvitis in patients having rheumatic fever in the absence of clinical evidence of cardiac involvement, and to evaluate its prognosis. METHODS AND RESULTS: Between November 1998 and September 1999, we identified 26 consecutive patients with silent valvitis in presence of rheumatic fever but in the absence of clinical signs of carditis. The patients, eight female and 18 male, were aged from 6 to 16 years, with a mean of 9.9+/-2.7 years. Major findings were arthritis in 16, chorea in 7, and arthritis and erythema marginatum in 1 patient. Two cases had arthralgia with equivocal arthritic signs and Doppler echocardiographic findings of pathologic mitral regurgitation. Silent pathologic mitral regurgitation was found in 12 cases, and aortic regurgitation in 2 cases. All patients with arthritic findings were treated with acetylsalicylic acid with one exception, this patient receiving both prednisone and acetylsalicylic acid. No antiinflammatory treatment was given to patients with chorea. After a mean follow up of 4.52 months, valvar regurgitation disappeared in 4 patients, including the one with migratory arthralgia and no other major criterions. All six patients with chorea and silent carditis still have mitral insufficiency. CONCLUSION: Acute rheumatic fever without clinical carditis is not a benign entity. Doppler echocardiographic findings of subclinical valvar insufficiency, therefore, should be considered as carditis when seeking to establish the diagnosis of acute rheumatic fever. PMID- 11388619 TI - A comparison of tissue Doppler imaging and velocities of transmitral flow in children with elevated left ventricular preload. AB - Compared with transmitral velocities of flow, myocardial wall velocities obtained by tissue Doppler imaging are less influenced by left atrial pressure. The data supporting this assumption, however, are limited in patients with congenital cardiac disease. The aim of this study was to compare the effects of left ventricular preload on transmitral inflow and velocities assessed by tissue Doppler imaging. Tissue Doppler imaging, and conventional Doppler echocardiography with simultaneous invasive hemodynamic studies, were performed in 33 patients with a simple ventricular septal defect or patency of the arterial duct. Transmitral velocities (E, A) and mitral annular velocities (Ea, Aa) were measured, permitting calculation of the ratio of E to Ea. The ratio of pulmonary to systemic flows, and mean left atrial pressure, were also measured. In 10 of 33 patients, echocardiographic and hemodynamic studies were performed 4 to 5 months after surgery. The E and A values in the patients were greater than those in the controls (p < 0.01). In contrast, neither Ea nor Aa differed between the two group. The ratio of E to Ea in the patients increased significantly compared with that in the controls (8.9+/-2.1 vs 7.3+/-1.3, p < 0.01). The E value was directly related to mean left atrial pressure and the ratio of pulmonary to systemic flows. The velocities measured by Tissue Doppler imaging, however, had no significant relationship to either of these measurements. The ratio of E to Ea correlated well with mean left atrial pressure (r = 0.75, p < 0.01). In 10 post operative patients, the values for E and A decreased from 119+/-14 to 89+/-10 cm/sec (p < 0.01) and from 91+/-22 to 61+/-9 cm/sec (p < 0.01), respectively. No significant changes were noted in the values of Ea and Aa. The ratio of E to Ea, and mean left atrial pressure, decreased from 10.3+/-1.9 to 8.2+/-1.5 (p < 0.01) and from 11.0+/-1.8 to 7.4+/-1.0 mmHg (p < 0.01), respectively. The percentage change in left atrial pressure correlated with the percent change in the ratio of E to Ea (r = 0.64, p < 0.05). The present study showed that the velocities obtained with tissue Doppler imaging are less dependent on mean left atrial pressure in children with elevated left ventricular preload caused by a left-to right shunt. PMID- 11388620 TI - Self-management of oral anticoagulation in children with congenital heart disease. AB - OBJECTIVE: The concept of self-management of oral anticoagulation has been shown to entail better quality of treatment than conventional management when assessed in selected adults. We have extended the concept of self-management to include children with congenital cardiac disease, hypothesizing self-management of oral anticoagulation is also possible in this subset of patients. Our aim was to assess the quality of self-management. METHODS: We trained 14 children aged from 2.2 to 15.6 years, with a mean age of 9.7 years, and their parents, in domiciliary analysis of the International Normalized Ratio and necessary adjustment of dosage of coumarin. The curriculum for training lasted for 27 weeks, and the patients and their parents were followed for a period of up to 31 months by weekly measurement of the values obtained for the International Normalized Ratio. RESULTS: The patients were observed over a mean of 547 days, with a range from 214 to 953 days. The patients were within the therapeutic targetted range of the International Normalized Ratio for a median of 65.5% of the time, with a range from 17.6% to 90.4%. None of the patients experienced thromboembolic or bleeding complications requiring doctoral intervention. All the patients and their parents expressed full satisfaction with the treatment. CONCLUSION: Self-management of oral anticoagulation provides a good quality of treatment, which is feasible and safe in selected children with congenital cardiac disease. PMID- 11388621 TI - Psychosocial functioning in children after transplantation of the heart, and heart and lungs. AB - We studied 10 boys, and 15 girls, all below the age of 16, who had been referred to the National Hospital in Norway for evaluation for transplantation of either the heart, or the heart and lungs. These represent the complete cohort of patients being considered for transplantation between 1990 and 1997. Of the 25 children and their families, 24 sets underwent a comprehensive psychosocial assessment, including interviews with both parents and their children. The parents completed the Child Behavior Checklist and the General Health Questionnaire. We had accepted 15 children for transplantation and placed them on the waiting list. The others were rejected for medical reasons, and 3 died whilst waiting for an organ. One was reconsidered for conventional surgery and removed from the list. Transplantation was performed in 11 children, whilst one of the patients we had rejected underwent transplantation abroad, and was included in the study. This left 12 patients in the final sample, with a mean age of 8 years, and with a range from 11 months to 13.9 years. We reassessed their psychosocial and physical functioning two years after transplantation. Of those undergoing transplantation of the heart and lungs, two were severely affected by progressive obliterative bronchiolitis. The others were in good general physical condition. At the assessment prior to transplantation, three already fulfilled the criterions for diagnosis of an overanxious disorder. Two others had symptoms of anxiety and depression, but without fulfilling the accepted criterions. At follow up, two patients retained this psychiatric diagnosis. Increased levels of stress were uncovered in the parents prior to surgery, but these had normalised at follow-up. The study shows that, in general, physical and psychological conditions improve in children undergoing transplantation, but they and their parents live in a stressful environment, and are in need of psychosocial support both before and after transplantation. PMID- 11388622 TI - Oxygen free radicals in children with acute rheumatic fever. AB - We have investigated the relationship between oxygen free radicals and acute rheumatic fever with regard to diagnosis of the disease process. At the time of diagnosis, we measured the levels of reactive oxygen molecules in the plasma, this being a parameter for oxygen free radicals, and discovered the levels to be significantly higher when compared with those measured in a control group (P<0.05). The levels measured in the plasma, however, were not statistically different among patients with and without carditis. We found a progressive decrease in the levels measured in the plasma when patients with acute rheumatic fever were tested on the 15th, 30th and 90th days subsequent to diagnosis. By the 90th day, levels measured in the plasma were still higher, but no longer significantly elevated, when compared with the control group. The present study is preliminary, but raises the possibility that measurement of oxygen free radicals in the plasma could be used as a laboratory test for active state of acute rheumatic fever. Further investigations will be needed, nonetheless, to determine the clinical application of this technique. PMID- 11388623 TI - Incidence and predictors for the development of significant supradiaphragmatic decompressing venous collateral channels following creation of Fontan physiology. AB - The occurrence of supradiaphragmatic decompressing venous collateral channels following construction of a bidirectional cavopulmonary connection or completion of the Fontan operation resulting in abnormal systemic hypoxemia has been infrequently described. In addition, the incidence and predictors of these channels have not been well delineated, especially in those patients without formation of such structures preoperatively. I evaluated, retrospectively, 40 patients who had undergone either construction of a bidirectional cavopulmonary shunt or completion of the Fontan operation, and who had complete pre and postoperative hemodynamic and angiographic data. Of the patients, 17 (43%) had developed a total of 21 decompressing venous collateral channels, of which 7 (18%) were considered to be hemodynamically significant requiring transcatheter coil occlusion. Of all variables examined, seven patients with significant decompressing collaterals had a greater transpulmonary gradient at follow-up catheterization (8 +/- 2 vs 5 +/- 2 mmHg, p=.01) and lower systemic saturations at routine clinical follow-up visits (82 +/- 5 vs 89 +/- 5 mmHg, p =.007) in comparison to the 33 others. When not evident preoperatively, decompressing venous collateral channels develop in a significant number of patients following conversion to Fontan physiology. If sufficiently large, they may produce lower than expected systemic saturations for the observed cardiac physiology. The larger decompressing channels are more likely to occur when a greater transpulmonary gradient exists postoperatively, which may require cardiac catheterization and transcatheter coil occlusion. PMID- 11388624 TI - Reduction in levels of triiodothyronine following the first stage of the Norwood reconstruction for hypoplastic left heart syndrome. AB - OBJECTIVE: Thyroid hormone has important effects on cardiovascular performance. This study was performed to evaluate the changes in levels of triiodothyronine following the first stage of reconstruction for hypoplastic left heart syndrome. METHODS: We enrolled 14 newborns with hypoplastic left heart syndrome scheduled for first stage reconstruction. Blood samples were obtained pre-, intra-, and post-operatively. Levels of free and total triiodothyronine were determined by radioimmunoassay. Statistical comparison was performed using Wilcoxon's signed rank test. RESULTS: The levels of free triiodothyronine decreased from a baseline of 355+/-31 pg/dl to 205+/-21 pg/dl upon the institution of bypass, and declined to a level of 135+/-9 pg/dl at 24 hours postoperatively. Similarly, levels of total triiodothyronine decreased from 101+/-15 ng/dl to 65+/-4 ng/dl upon the institution of bypass, and continued to decline during the first 24 hours postoperatively. Levels of free and total triiodothyronine had returned to baseline by the fifth postoperative day. CONCLUSIONS: The data demonstrate significant decreases in levels of free and total triiodothyronine during the early postoperative period. These changes in levels of thyroid hormone may have adverse effects on cardiac function during this phase of recovery. PMID- 11388625 TI - A pilot study of expenditures on, and utilization of resources in, health care in adults with congenital heart disease. AB - BACKGROUND: Congenital cardiac disease may be a chronic condition, necessitating life-long follow-up for a substantial proportion of the patients. Such patients, therefore, are often presumed to be high users of resources for health care. Information on utilization of resources in adults with congenital heart disease, however, is scarce. METHODS: This retrospective pilot study, performed in Belgium, investigated 192 adults with congenital heart disease to measure the annual expenditures and utilization of health care and compared the findings with data from the general population. We also sought to explore demographic and clinical parameters as predictors for the expenditures. RESULTS: Hospitalization was documented in 20.3% of the patients, with a median length of stay of 5 days. The overall payment by health insurance associations in 1997 was 1794.5 ECU per patient, while patients paid on average 189.5 ECU out-of-pocket. For medication, the average reimbursement and out-of-pocket expenses were estimated at 78 ECU and 20 ECU, respectively. Expenditures for patients with congenital heart disease were considerably higher than the age and gender-corrected expenditures for the general population (411.7 ECU), though this difference was accounted for by only one-eighth of the cohort of those with congenital heart disease. In general, higher expenditures were associated with abnormal left ventricular end-diastolic diameter, female gender, functional impairment and higher age, although the explained variance was limited. CONCLUSION: Our study has provided pilot data on the economic outcomes for patients with congenital heart diseases. We have identified parameters that could predict expenditure, but which will have to be examined in future research. This is needed to develop guidelines for health insurance for those with congenital heart diseases. PMID- 11388626 TI - Feasibility of transcatheter closure of multiple defects within the oval fossa. AB - BACKGROUND: Multiple perforations in the floor of the oval fossa may be an obstacle for transcatheter closure. Thus, we analyzed the interventions in 33 patients with more than one interatrial communication in comparison with 370 procedures with a single defect. METHODS AND RESULTS: A diagnostic catheterization, which included a balloon-sizing maneuver, was performed. We implanted a total of 46 occluders, made up of 42 Amplatzers and 4 CardioSEALs. In 20 patients, the defects were closed with a single occluder, namely 18 Amplatzer and 2 CardioSEAL devices. Complete closure was achieved in 15 patients, while a tiny residual shunt remained in 5 patients. In 13 patients, two devices were implanted, without any residual shunt being found immediately after implantation. In 3 patients, the occluders did not touch each other. In 10 patients, their rims overlapped. In comparison with the control group, the group with multiple defects did not differ in the distribution of age, gender, and indications for device closure. The mean time of the procedure, and the time required for fluoroscopy, however, were significant longer (P<0.001). These times ranged from 45 to 250 minutes with a median of 140 minutes, and from 0.0 to 39.2 minutes, with a median of 12.0 minutes, respectively. Also, the association with an atrial septal aneurysm was significantly more frequent (61 vs. 17%; P<0.001). The times taken during insertion of double devices were also significantly longer than those needed for insertion of a single device (P<0.001). CONCLUSIONS: Transcatheter closure of multiple defects within the oval fossa is feasible with currently available occluders, albeit than, in selected cases it is necessary to implant two devices. PMID- 11388627 TI - Late anastomotic stenosis after correction of totally anomalous pulmonary venous connection. PMID- 11388628 TI - Maria Victoria de la Cruz (1916-1999). PMID- 11388629 TI - Continuing medical education. Double outlet right ventricle. PMID- 11388630 TI - The bronchial cast syndrome after the fontan procedure: further evidence of its etiology. AB - We describe a three-year-old boy who presented with recurrent expectoration of bronchial casts six months following creation of a fenestrated lateral tunnel Fontan circulation for pulmonary atresia with intact ventricular septum. Cardiac catheterization demonstrated elevated central venous pressure with two areas of stenosis within the Fontan circuit, specifically at the junction of the right superior caval vein and the right pulmonary artery, and between the atrial baffle and the right superior caval vein. Insertion of Palmaz stents in these areas resulted in a reduction in central venous pressure, and a transient reduction in production of casts. Eight weeks after catheterization, however, he produced further casts, which resulted in hypoxia, respiratory arrest and death. We reviewed the autopsied specimens obtained from patients with the Fontan circulation over an eleven-year period at our institution in order to ascertain the prevalence of subclinical production of bronchial casts. We found no casts in the thirteen patients examined. Hemodynamic assessment is vital in all patients who develop this syndrome, and should be the primary focus of investigation, rather than solely directing efforts at lysis of casts. PMID- 11388631 TI - Pulmonary atresia with intact ventricular septum, right-sided aortic arch and ventriculocoronary connection--prenatal echocardiographic diagnosis. AB - The authors describe the prenatal echocardiographic diagnosis of a rare case of pulmonary atresia with intact ventricular septum, right-sided aortic arch, and ventriculocoronary connection in a fetus at 21 weeks gestation. The diagnosis was confirmed at autopsy. PMID- 11388632 TI - The role of tissue plasminogen activator in the successful treatment of infected cardiac thrombus in children. AB - Infected cardiac thrombus is rare in children, with antibiotic or antifungal agents used as the first line of treatment. Persistence is an indication for surgical intervention. We describe two children who were treated successfully with a combination of antibiotic and antithrombotic agents. Use of antithrombotic agents promotes degeneration of fibrin, thus reducing the mass and facilitating the diffusion of the antibiotic and/or antifungal agents. PMID- 11388633 TI - Major intrahepatic veno-venous fistula after modified Fontan operation treated by transcatheter implantation of amplatzer septal occluder. AB - Patients with complex congenital cardiac malformations who have been converted to the Fontan circulation with partial exclusion of the hepatic veins may develop progressive cyanosis because of formation of intrahepatic veno-venous malformations. We describe transcatheter closure of a major intrahepatic fistula in such a setting using an Amplatzer septal occluder delivered by the left jugular venous approach in a 5 year old boy. PMID- 11388634 TI - Biventricular repair in a patient having double outlet right ventricle with unusual straddling of the mitral valve. AB - The authors achieved biventricular repair in a 1-year-old boy having double outlet right ventricle and unusual straddling of the mitral valve. There was a substantial abnormal muscular bar present within the right ventricle, and some of the tendinous cords of the mitral valve were attached to this structure. Preoperative echocardiography was particularly informative in enabling the surgeon to arrange the optional operative procedures. The postoperative course was smooth and uneventful. PMID- 11388635 TI - Anomalous extension of ductal tissue within the pulmonary artery. PMID- 11388636 TI - Molecular pharmacology of T-type Ca2+ channels. AB - Over the past few years increasing attention has been focused on T-type calcium channels and their possible physiological and pathophysiological roles. Efforts toward elucidating the exact role(s) of these calcium channels have been hampered by the lack of T-type specific antagonists, resulting in the subsequent use of less selective calcium channel antagonists. In addition, the activity of these blockers often varies with cell or tissue type, as well as recording conditions. This review summarizes a variety of compounds that exhibit varying degrees of blocking activity towards T-type Ca2+ channels. It is designed as an aid for researchers in need of antagonists to study the biophysical and pathological nature of T-type channels, as well as a starting point for those attempting to develop potent and selective antagonists of the channel. PMID- 11388637 TI - Chloride channels and their functional roles in smooth muscle tone in the vasculature. AB - Although evidence of important contributions by Cl- channels to agonist-induced currents have been reported in vascular smooth muscle cells, the functional roles played by Cl- channels in the smooth muscle contraction and in setting the membrane potential remain essentially obscure. All of the admittedly few papers published have focused on the physiological roles of Cl- channels in the contraction and membrane depolarization elicited by agonists. At present, it seems likely that in vascular cells: a) Cl- conductance contributes to membrane depolarization, with the subsequent contraction being due to Ca2+ release from the intracellular store sites, and b) Cl- movements through the membrane of the Ca2+ store sites also regulate Ca2+ release and Ca2+ uptake from/into the store sites. As a Ca2+-dependent Cl- current is most easily demonstrated under quasi physiological conditions (by the perforated patch-clamp method), the contribution made by Cl- channels to smooth muscle function may be more important than previously thought. The development of the new, selective Cl--channel blockers as well as the identification and gene engineering of the channel molecules are essential if we are to advance our knowledge of the physiology and pharmacology of the Cl- channels residing in vascular smooth muscle cells. PMID- 11388638 TI - Role of reduced glutathione and nitric oxide in the black tea extract-mediated protection against ulcerogen-induced changes in motility and gastric emptying in rats. AB - The aim of the present study was to investigate the underlying mechanism of the role of hot water extract of black tea [Camellia sinensis (L). O. Kuntze Theaceae] in normalizing the changes in intestinal transit and gastric emptying induced by various ulcerogenic agents in experimental rats. Intestinal transit as well as gastric emptying were significantly reduced in rats treated with glutathione (GSH) depleting agents, diethyl maleate (DEM), indoacetamide (IDA) and N-ethyl maleimide (NEM). Prior oral administration of black tea extract (BTE) at 20 ml/kg of a 10% solution, i.g. once a day for 7 days significantly increased the intestinal transit and gastric emptying with restoration of serum GSH level. Singular administration of succimer (60 mg/kg, i.g.), the standard sulfhydryl containing antiulcer agent used as a reference drug, was also effective. Increase in intestinal transit caused by BTE was reversed both by N-omega-nitro-L-arginine methyl ester (L-NAME) (25 mg/kg, i.p.) and N-omega-monomethyl-L-arginine (L-NMMA) (25 mg/kg, i.p.), but not with N-omega-nitro-D-arginine methyl ester (D-NAME) (25 mg/kg, i.p.). Furthermore, restoration of intestinal nitric oxide synthase (NOS) activity was found to be associated with BTE treatment. These results provide evidence that nitric oxide may play a role in BTE-mediated improvement of intestinal motility changes and gastric emptying induced by DEM, IDA and NEM. PMID- 11388639 TI - Angiotensin-converting enzyme inhibitors and AT1-receptor antagonist restore nitric oxide synthase (NOS) activity and neuronal NOS expression in the adrenal glands of spontaneously hypertensive rats. AB - During development of hypertension in spontaneously hypertensive (SHR) rats, the activity of adrenal nitric oxide synthase (NOS) was investigated. SHR and Wistar Kyoto (WKY) rats were studied at different ages: 3-4, 7-8 and 12-13 weeks after birth. Basal NOS activity was measured by the ability of homogenate to convert [3H]-L-arginine to [3H]-L-citrulline. At all ages, SHR rats exhibited 50-60% reduction in NOS activity when compared to age-matched WKY rats. In a following study, SHR rats (12-13 weeks) were treated chronically with the angiotensin I converting enzyme inhibitors (ACE-I) captopril or enalapril, or the AT1-receptor antagonist losartan (2 x 25, 10 and 60 mg/kg per day for 10 days, respectively). The total NOS activity and protein expression of NOS isoenzymes from adrenals were determined. The basal NOS activity and protein expression of neuronal NOS (nNOS) was significantly increased in treated SHR rats when compared to control rats. The isoforms endothelial NOS and inducible NOS were undetectable. We conclude that impaired NO synthesis in the adrenal glands of SHR rats may contribute to the onset and maintenance of hypertension. The upregulation of nNOS protein in the adrenal glands may be one of the mechanisms by which ACE inhibitors and AT1-receptor antagonists by restoring the NO synthesis, mediate their antihypertensive effects. PMID- 11388640 TI - Blocking effect of bepridil on Na+/Ca2+ exchange current in guinea pig cardiac ventricular myocytes. AB - We examined the effect of bepridil, a class IV antiarrhythmic drug, on Na+/Ca2+ exchange current (I(NCX)) in single guinea pig cardiac ventricular cells using the whole-cell voltage clamp technique. I(NCX) was recorded by ramp pulses from the holding potential of -60 mV in the presence of 140 mM Na+ and 1 mM Ca2+ in the external solution and 20 mM Na+ and 119 nM free Ca2+ (7 mM Ca2+ and 20 mM BAPTA) in the internal solution. Bepridil suppressed I(NCX) in a concentration dependent manner. The IC50 value was 8.1 microM with a Hill coefficient of 0.8. Intracellular treatment with trypsin via the pipette solution attenuated the blocking effect of bepridil, suggesting that the inhibitory site is on the cytosolic side of the Na+/Ca2+ exchanger. In the absence of albumin in the external solution, 10 microM bepridil inhibited I(NCX) by 46+/-7% (n = 8), while bepridil blocked it by 28+/-8% (n = 6) in the presence of albumin. Bepridil inhibited I(NCX) in a supra-therapeutic concentration range. PMID- 11388641 TI - Extracellular ATP potentiates steroidogenic effect of adrenocorticotropic hormone in bovine adrenocortical fasciculata cells. AB - We examined the effect of extracellular adenosine 5'-triphosphate (ATP) on adrenocorticotropic hormone (ACTH)- and angiotensin II-induced steroidogenesis in bovine adrenocortical fasciculata cells. The low concentration of ATP (5 microM) potentiated ACTH-induced steroidogenesis synergistically. However, the purine derivative did not affect angiotensin II-induced steroidogenesis. Although adenosine (100 microM) (a metabolite of ATP) showed a weak steroidogenic effect, it did not potentiate ACTH-induced steroidogenesis. ATP also enhanced the steroidogenesis by NaF synergistically in bovine adrenocortical cells, but did not potentiate forskolin- and dibutyryl cyclic AMP-induced steroidogenesis. The stimulating effect of ACTH on cyclic AMP production was synergistically accelerated by ATP (5 microM), which has no effect by itself on cyclic AMP formation. These results suggest that extracellular ATP affected the ACTH receptor-adenylyl cyclase coupling processes, and potentiation of steroidogenesis by ACTH ensued in bovine adrenocortical fasciculata cells. PMID- 11388642 TI - Imaging of Ca2+ release by caffeine and 9-methyl-7-bromoeudistomin D and the associated activation of large conductance Ca2+-dependent K+ channels in urinary bladder smooth muscle cells of the guinea pig. AB - Ca2+ release by caffeine and 9-methyl-7-bromoeudistomin D (MBED) and the concomitant activation of large conductance Ca2+-dependent K+ (BK) channels were analyzed using confocal Ca2+ imaging and whole cell voltage-clamp methods in guinea pig urinary bladder smooth muscle cells. Puff application of 3 or 10 mM caffeine for several seconds (2 - 5 s) elicited a large increase in intracellular Ca2+ concentration ([Ca2+]i) and induced a phasic outward current at a holding potential of -40 mV. The phasic outward current was the summation of spontaneous transient outward currents (STOCs) due to marked activation of BK channels and was followed by a short cessation of STOCs. Although the increase in superficial [Ca2+]i by caffeine was faster than that in global [Ca2+]i, the peak [Ca2+]i was identical in these areas. Puff application of 100 microM MBED also markedly enhanced STOCs for a few seconds. This response to MBED was not observed when stored Ca2+ was depleted by caffeine. The increase in [Ca2+]i by MBED occurred mainly in superficial areas. Longer application of 100 microM MBED for 2 min did not induce significant global [Ca2+]i increase but decreased the amount of Ca2+ release and cell shortening during the subsequent application of 10 mM caffeine. These results indicate that short application of MBED releases Ca2+ preferentially from superficial storage sites, presumably due to its slow approach to deeper sites. MBED may be a good pharmacological tool to manipulate selectively the superficial Ca2+ stores related to STOCs. PMID- 11388643 TI - Gentamicin decreases the abundance of aquaporin water channels in rat kidney. AB - The present study was performed to examine whether the gentamicin-induced urinary concentration defect is related to an altered regulation of aquaporin (AQP) water channels in the kidney. Male Sprague-Dawley rats were subcutaneously injected with gentamicin (20, 50 or 100 mg/kg per day) for 6 days. The protein expression of AQP1-3 channels and the catalytic activity of adenylyl cyclase were determined in the kidney. Gentamicin treatment resulted in renal failure associated with decreased tubular free water reabsorption and increased urinary flow rate. The expression of AQP2 proteins was significantly decreased in the kidney, in which the cortex was most susceptible, followed by the outer medulla and inner medulla in order. Gentamicin treatment also decreased the shuttling of AQP2, as evidenced by a decrease of its expression in the membrane fraction in proportion to that in the cytoplasmic fraction. The protein expression of AQP1 as well as that of AQP3 was also decreased in the cortex by treatment with the highest dose of gentamicin. The cAMP generation in response to arginine vasopressin or sodium fluoride was decreased by gentamicin, while that to forskolin was not significantly altered. These findings suggest that the primary impairment in the pathway leading to the generation of cAMP lies at the level of G proteins, resulting in a decreased expression of cAMP-mediated AQP channels. The gentamicin induced urinary concentration defect may in part be accounted for by a reduced abundance of AQP water channels in the kidney. PMID- 11388644 TI - Inhibitory potential of herbal medicines on human cytochrome P450-mediated oxidation: properties of umbelliferous or citrus crude drugs and their relative prescriptions. AB - To investigate the possible drug interaction with herbal medicine, hot water decoctions or 40% ethanol infusions of several Umbelliferous or Citrus crude drugs and their prescriptions were examined in vitro for their abilities to inhibit human cytochrome P450 3A (CYP3A). Addition of each decoction or infusion from Baizhi (Angelica dahurica and varieties), Qianghuo (Notopterygium incisum or N. forbesii), Duhuo (Angelica biserrata), Fangfeng (Saposhnikovia divaricata), Danggui (Angelicasinensis), Zhishi or Zhiqiao (Citrus aurantium) resulted in various degrees of human CYP3A inhibition as determined by microsomal testosterone 6beta-hydroxylation. The inhibitory potency was consistent with the abundance of the hydrophobic components for each sample. Experiments on the infusion of a Japanese Baizhi (BZ1) showed the major role of furanocoumarins on human CYP3A inhibition. Some of the crude drugs and a related prescription showed increased inhibition after the preincubation, suggesting the involvement of a mechanism-based inhibition. Some formulated prescriptions, however, showed intense inhibition with their hydrophobic fractions rather than with their hydrophobic fractions, suggesting that components other than furanocoumarins in herbal prescriptions may also cause CYP3A inhibition. These results indicate the necessity of intensive investigations on the possible drug interaction with traditional medicines. PMID- 11388645 TI - Comparison of cedar pollen-induced allergic rhinitis in passively and actively sensitized guinea pigs. AB - We have developed an allergic rhinitis model in guinea pigs using Japanese cedar pollen as antigen. In the present study, we examined whether provocation by pollen induces similar magnitudes of rhinitis symptoms in passively and actively sensitized guinea pigs. One group of animals was actively sensitized by intranasal application of pollen extract, and another was passively sensitized by intraperitoneal injection with anti-pollen serum. Actively and passively sensitized groups were then challenged by repeated and a single pollen inhalation, respectively. In both groups, sneeze was induced immediately after the challenge. The actively sensitized animals developed not only early but also late nasal blockage, whereas the passively sensitized animals showed only early nasal blockage. In both groups, an H1 antagonist, mepyramine, inhibited the occurrence of sneezing but did not inhibit nasal blockage. Nasal hyperresponsiveness to intranasal instillation of leukotriene D4 was obvious only in the actively sensitized animals. We thus conclude that although early nasal blockage is induced by a single antigen-antibody reaction, repetitive anaphylactic reaction is required for occurrence of late nasal blockage and hyperresponsiveness to stimuli. Furthermore, histamine plays a central role in induction of sneezing but not in nasal blockage, irrespective of whether animals are actively or passively sensitized. PMID- 11388646 TI - Involvement of angiotensin II in progression of renal injury in rats with genetic non-insulin-dependent diabetes mellitus (Wistar fatty rats). AB - Wistar fatty (WF) rats have a genetic predisposition to hyperglycemia, polyuria, hyperinsulinemia, hyperlipidemia, obesity and nephropathy. These phenotypic characteristics are similar to those observed in obese patients with non-insulin dependent diabetes mellitus (NIDDM) nephropathy. In this study, the effects of two types of renin-angiotensin system inhibitors, an angiotensin II type 1 receptor antagonist (AT1A) and an angiotensin I-converting enzyme inhibitor (ACEI), on renal injury in WF rats were studied during the progressive phase of diabetic nephropathy. An AT1A, candesartan cilexetil (1 mg/kg), and an ACEI, enalapril (10 mg/kg), were administered orally once a day for 12 weeks, beginning when the rats were 27-week-old and already showed diabetic nephropathy and obesity. Both drugs prevented an increase in proteinuria during the experimental period. Furthermore, after 4-week intervention, the levels of proteinuria were markedly lower in drug-treated rats. At the end of the experiment, both drugs prevented the development of glomerular lesions without affecting glucose metabolism and obesity. In conclusion, the inhibition of angiotensin II activity ameliorated both existing proteinuria and the progression of proteinuria, resulting in preservation of glomerular structure. Thus angiotensin II plays important roles in the development and the progression of nephropathy in genetically obese diabetic WF rats. PMID- 11388648 TI - Altered susceptibility to ischemia-reperfusion injury in isolated-perfused hearts of short-term diabetic rats associated with changes in non-enzymatic antioxidants. AB - The effects of short-term (2-week) diabetes on myocardial ischemia-reperfusion (I R) injury and associated changes in myocardial non-enzymatic antioxidant level were examined. Isolated-perfused hearts prepared from control and diabetic rats were subjected to increasing periods of ischemia and reperfusion, and myocardial I-R injury was assessed by measuring the extent of lactate dehydrogenase (LDH) leakage and contractile force recovery. While a brief period (20 min) of post ischemic reperfusion caused a smaller extent of LDH leakage, the prolonged period (40 min) of reperfusion produced a greater degree of I-R injury in diabetic hearts, as indicated by the impaired recovery of contractile force. The apparent protection against I-R injury in diabetic hearts during the early phase of post ischemic reperfusion was associated with increases in myocardial reduced glutathione/ascorbic acid and a-tocopherol levels, with the effect on a tocopherol being most prominent. Insulin treatment could reverse the diabetes associated changes in susceptibility to myocardial I-R injury and antioxidant response. The ensemble of results indicates that the myocardium isolated from short-term diabetic rat can produce a beneficial antioxidant response to I-R challenge, which may, in turn, be attributable to the decreased susceptibility to I-R injury observable during the early phase of reperfusion. PMID- 11388647 TI - Pharmacological properties of R-755, a novel acyl-CoA:cholesterol acyltransferase inhibitor, in cholesterol-fed rats, hamsters and rabbits. AB - R-755 (N-(2,6-diethylphenyl)-N'-[3-(2-methylphenyl)-6,7-dihydro-5H cyclopenta[f[l]benzothiophen-2-yl]urea), a novel acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor, has been characterized in vitro, ex vivo and in vivo. R-755 potently inhibited ACAT activities, with IC50 values from 2.5 to 64 nM, in rabbit intestinal microsomes and several cell lines (CaCo-2, THP-1 and J774A.1 cells). R-755 reduced serum cholesterol and triglyceride levels and liver cholesterol contents in cholesterol-fed rats, hamsters and rabbits. Rabbits were fed a high cholesterol diet for 2 weeks and further fed the same diet containing R-755 for 2 weeks. R-755 dose-dependently reduced cholesterol content and ACAT activity in the aorta. When phorbol 12-myristate 13-acetate-treated THP-1 and J774A.1 cells were incubated in the medium containing 20% of serum from rats administered R-755, the ACAT activities of the cells were inhibited. Rabbits were fed a high cholesterol diet for 8 weeks to establish aortic atherosclerosis and then fed a normal diet with or without R-755 for 8 weeks. R-755 dose-dependently reduced the surface area with atherosclerotic involvement and cholesterol contents in the aorta, although plasma cholesterol level did not differ from that in the control group. These results suggest that R-755 is a potent hypolipidemic agent and has a direct antiatherosclerotic activity at the arterial wall. PMID- 11388649 TI - D-galactose-specific sea urchin lectin sugar-specifically inhibited histamine release induced by datura stramonium agglutinin: differences between sugar specific effects of sea urchin lectin and those of D-galactose- or L-fucose specific plant lectins. AB - A new sea urchin lectin from Toxopneustes pileolus, is D(+)galactose (Gal)-, D(+)fucose (Fuc)-specific. Incubation of rat peritoneal mast cells with the lectin in the presence of 0.3 mM CaCl2 for 10 min significantly and dose dependently inhibited the histamine release induced by N-acetyl glucosamine (GlcNAc)-specific Datura stramonium agglutinin (DSA), an activator of the Gi protein-dependent pathway in mast cells. This inhibition by the sea urchin lectin was sugar-specifically reversed in the presence of D(+)Gal or D(+)Fuc but not L( )Fuc. The sea urchin lectin had no effect on the histamine release induced by compound 48/80, slightly inhibited the histamine release induced by substance P and mastoparan, and slightly enhanced the histamine release induced by melittin, but these effects were not dose-dependent. Compound 48/80, substance P, mastoparan and melittin are mast cell activators without sugar residues. It is suggested that the lectin binds to D(+)Gal residues of DSA to interfere with mast cell activation induced by DSA, a glycoprotein with arabinose and Gal residues. The effects of plant lectins with affinity to D(+)Gal, N-acetyl galactosamine and/or sialic acid and L(-)Fuc on the histamine release induced by DSA, compound 48/80 and substance P were also examined. PMID- 11388650 TI - Effects of olopatadine hydrochloride on the increase of histamine and peptide leukotrienes concentrations in nasal lavage fluid following the antigen-antibody reaction in actively sensitized guinea pigs. AB - To investigate the mechanism for the amelioration by olopatadine hydrochloride (olopatadine) of allergic rhinitis, we determined its effects on the increase of chemical mediator concentrations in nasal lavage fluid following the intranasal antigen challenge in guinea pigs actively sensitized with DNP-Ascaris. The concentrations of histamine and peptide-leukotrienes increased 10 min after the challenge. Olopatadine at 10 mg/kg (p.o.) significantly prevented the increase of histamine and tended to inhibit the increase of peptide-leukotrienes. The inhibition by olopatadine of the nasal symptoms seems to involve the inhibitory effect on the releases of histamine and, possibly, p-LTs into the nasal cavity. PMID- 11388651 TI - Distribution of cellular prion protein in normal human cerebral cortex--does it have relevance to Creutzfeldt-Jakob disease? AB - Creutzfeldt-Jakob disease and bovine spongiform encephalopathy are the best known forms of prion diseases. A basis for their pathogenesis is the transformation of normal prion protein to abnormal prion protein. This would mean that either loss of normal function or a gain of a toxic function of the prion protein would play a major role. Since the prime target for Creutzfeldt-Jakob disease in humans is the neocortex, and the intracortical distribution of the destructive process in prion diseases appears not to be haphazard, it may be that a clear cortical study of normal prion protein production in the premorbid human neocortex might contribute to insight in the pathogenesis of prion diseases. As no such study is available, we performed a detailed study in normal human cortex using immunohistochemistry for prion protein, in both frozen and vibratomised tissue, and in situ hybridisation for prion protein mRNA. We have found normal prion protein production mainly in the upper cortical neurons in neocortex and Purkinje cells in the cerebellum. This finding implicates that normal prion protein is more important as an anti-apoptotic signal in disease than abnormal prion protein is as a toxic substance. PMID- 11388652 TI - Caspase-3 apoptotic signaling following injury to the central nervous system. AB - Apoptotic cell death is a fundamental and highly regulated biological process in which a cell is instructed to participate actively in its own demise. This process of cellular suicide is activated by developmental and environmental cues and normally plays an essential role in eliminating superfluous, damaged, and senescent cells of many tissue types. In recent years, a number of experimental studies have provided evidence of widespread neuronal and glial apoptosis following injury to the central nervous system (CNS). These studies indicate that injury-induced apoptosis can be detected from hours to days following injury and may contribute to neurological dysfunction. Given these findings, understanding the biochemical signaling events controlling apoptosis is a first step towards developing therapeutic agents which would target this cell death process. This review will focus on the molecular cell death pathways responsible for generating the apoptotic phenotype, summarize what is currently known about apoptotic signals activated in the injured CNS, and what potential strategies might be pursued to reduce this cell death process as a means to promote functional recovery. PMID- 11388653 TI - Parkinson's disease and other alpha-synucleinopathies. AB - Parkinson's disease is the most common movement disorder and the second most common neurodegenerative disease. Neuropathologically, it is characterized by the degeneration of nerve cells that develop filamentous inclusions in the form of Lewy bodies and Lewy neurites. Recent work has shown that rare, familial forms of Parkinson's disease are caused by missense mutations in the alpha-synuclein gene and that the filamentous lesions of Parkinson's disease are made of alpha synuclein. The same is true of the Lewy body pathology that is associated with other neurodegenerative diseases, such as dementia with Lewy bodies. The filamentous inclusions of multiple system atrophy have also been found to be made of alpha-synuclein, thus providing an unexpected molecular link with Lewy body diseases. Recombinant alpha-synuclein assembles into filaments with similar morphologies to those found in the human diseases and with a cross-beta diffraction pattern characteristic of amyloid. The related proteins beta synuclein and gamma-synuclein are poor at assembling into filaments. They are not present in the pathological filamentous lesions and have not been found to be linked to genetic disease. The new work has established the alpha synucleinopathies as a major class of neurodegenerative disease. PMID- 11388654 TI - Aggregation-Dependent interaction of the Alzheimer's beta-amyloid and microglia. AB - Chronic glial activation possibly plays a role in chronic neurodegeneration in Alzheimer's disease (AD). It has been shown that amyloid peptide is capable of activating microglial cells in vitro. The aim of this study was to further characterize the structural preconditions for amyloid peptide in order to activate glial cells and to investigate whether this peptide is also able to induce glial activation in the living brain. We observed that amyloid peptide induced strong cellular activation in primary microglial cell culture as detected by the release of stable metabolites of nitric oxide (NO), when the peptide was fibrillar. For this activation, co-stimulation with interferon-gamma was a precondition. Using microdialysis of the living brain in a rat we observed pronounced NO generation when fibrillar amyloid peptide was stereotaxically injected. Non-fibrillar amyloid peptide did not induce such a glial reaction. No administration of interferon-gamma or any other co-stimulatory factor was necessary in vivo. Thus, we show that fibrillar, but not non-fibrillar amyloid peptide induced glial activation also in vivo. In the case of the living brain, the presence of deposits of fibrillar amyloid peptide could maintain a chronic microglial activation, ultimately leading to the progressive neurodegeneration associated with Alzheimer's disease. PMID- 11388655 TI - beta-Amyloid-induced cytotoxicity, peroxide generation and blockade of glutamate uptake in cultured astrocytes. AB - beta-Amyloid (betaA) is cytotoxic to neurons in culture by increasing hydrogen peroxide and altering calcium homeostasis. We have evaluated betaA-induced cytotoxicity, peroxide generation and glutamate (Glu) uptake in cultured astrocytes. Twenty-four hours after a single addition of either betaA25-35 or betaA1-40there was a concentration-dependent decrease in viability. Catalase or vitamin E showed no protective effect against betaA25-35 Dithiothreitol (DTT), N acetylcysteine (NAC) and cyclosporine A significantly prevented the toxic effects of both betaA25-35 and peroxide, while inhibition of peroxide detoxifying enzymes enhanced toxicity. Exposure to betaA25-35 or betaA1-40 increased peroxides at 2 h and 24 h, which was prevented by DTT and NAC, but not vitamin E. betaA25-35 inhibited Glu uptake in astrocytes and neurons in culture. Following exposure of neurons to betaA for 24 h there was decreased uptake and increased Glu levels in the culture medium, that resulted in gradual excitotoxicity. PMID- 11388656 TI - Protein S-100B: a serum marker for ischemic and infectious injury of cerebral tissue. AB - The S-100B protein is released by injured astrocytes. After passage through a disintegrated blood-brain barrier (BBB) the molecule can be detected in the peripheral circulation. We investigated the association between the extent of brain injury and S-100B concentration in serum in cerebral injury caused by cerebral ischemia and cerebral fungal infection. Study I: The S-100B serum concentration was serially determined in 24 patients with ischemic stroke at 4, 8, 10, 24, 72 hours after the onset of symptoms. We observed that patients with brain lesions larger than 5 cm3 exhibited significantly increased serum levels of S-100B at 10, 24 and 72 hours compared to those with lesion volumes below 5 cm3. Furthermore, an association between S-100B serum concentration and neurological outcome was observed. Study II: In a mouse model of systemic fungal infection with Candida albicans we observed that serum levels of S-100B increased at day 1 after intravenous infection. At this time we could histologically demonstrate brain tissue injury by invading hyphae which had crossed the BBB. Furthermore, reactive astrogliosis was demonstrated by immunohistochemistry. On day 7 we found a significant decrease of S-100B serum level compared to day 1 and 4. This was associated with a demarcation of the fungi with leukocytes in brain tissue at this late phase of infection. No further invasion through the BBB was seen on day 7. In conclusion, serum levels of S-100B reflect the time course of tissue injury in cerebral ischemia and cerebral infection to a similar extent. Thus, S-100B may be a useful marker to assess cerebral tissue injury. PMID- 11388657 TI - Reporting cerebrospinal fluid data: knowledge base and interpretation software. AB - The compilation of cerebrospinal fluid (CSF) patient data together with a graphic display of immunoglobulin patterns in a single CSF report has two main advantages: analytical and clinical plausibility control of a complex set of data improves quality assessment and allows improved clinical specificity and sensitivity for recognition of disease-related "typical" data patterns. The widespread use of automated on-line evaluation programs can now be combined with knowledge-based programs for interpretation by clinical chemists and neurologists. These programs are based on knowledge of neuroimmunology, blood-CSF barrier function and dysfunction, influence of CSF flow on concentrations of blood-derived and brain-derived proteins in CSF, specific intrathecal antibody synthesis and relevance of brain proteins for differential diagnosis of degenerative diseases. The relevance of hyperbolic discrimination functions in quotient diagrams for the detection of intrathecal immunoglobulin synthesis is compared with earlier, still frequently used, linear interpretation functions. Differences found in commercially available interpretation software are discussed. PMID- 11388658 TI - The intrathecal humoral immune response: laboratory analysis and clinical relevance. AB - In normal conditions, albumin and immunoglobulin (Ig)G in the cerebrospinal fluid (CSF) originate from the blood, and there is no antibody production within the central nervous system. Up to 20% of CSF proteins are intrathecally synthesized, but the major fraction is blood-derived. The CSF/serum albumin quotient (QAlb) is the best marker of the blood-CSF barrier function. The corresponding immunoglobulin quotients (QIGG, QIGA, QIGM) are not linearly related to QAlb and their correlations are defined by an hyperbolic equation. This equation is used to discriminate between a blood-derived and a locally produced fraction of immunoglobulins in case of an intrathecal humoral immune response. The detection of CSF-specific oligoclonal IgG is more sensitive than the quantitative comparison between QIGG and QAlb. A further step is the determination of antibody indices and the detection of specific oligoclonal antibodies by antigen-driven immunoblots. CSF analysis remains a cornerstone for the diagnosis of various neurological disorders, including multiple sclerosis and infectious diseases of the central nervous system. PMID- 11388659 TI - Source of endothelin-1 in subarachnoid hemorrhage. AB - Endothelin-1 is the most potent vasoconstrictor known to date. This peptide is believed to play a pathophysiological role in the development of vasospasm, the most important complication of subarachnoid hemorrhage (SAH). In the present study we investigated the release of endothelin-1 in SAH and analyzed the cellular source of this peptide. At a protein and mRNA level we were able to show that endothelin-1 is produced by mononuclear leukocytes. Complementary in vitro studies revealed that aging and subsequent hemolysis of blood is sufficient to induce production of endothelin-1 by mononuclear leukocytes. Thus, cerebrospinal fluid-derived mononuclear leukocytes are a source of endothelin-1 in patients suffering from SAH. This finding may have important therapeutic implications as anti-leukocyte strategies could prevent cerebrovascular complications in SAH patients. PMID- 11388660 TI - Polymorphism of apoprotein E (APOE), methylenetetrahydrofolate reductase (MTHFR) and paraoxonase (PON1) genes in patients with cerebrovascular disease. AB - Although controversial, data on the genetic polymorphism of apoprotein E (APOE), methylenetetrahydrofolate (MTHFR) and paraoxonase (PON1) genes implicate their role in the development of cerebrovascular disease. The aim of this study was to assess the association of polymorphism of APOE, MTHFR and PON1 genes in 56 stroke and 36 carotid stenosis patients, and in 124 control subjects by PCR-restriction fragment length polymorphism analysis. In the stroke group a significantly different MTHFR genotype distribution (p=0.004, odds ratio for T/T of 17.571), but no significant difference in APOE and PON1 allele and genotype distribution compared to the control was found. The carotid stenosis group exhibited a significantly different APOE allele and genotype distribution (p=0.023, odds ratio APOEepsilon3epsilon4 of 4.24), but no significant difference in the MTHFR and PON1 allele and genotype distribution from the control group. The preliminary results obtained in this study revealed an association of the MTHFR and APOE gene polymorphism with cerebrovascular disease, suggesting a significant risk for stroke in subjects who are homozygous for the T allele and for carotid stenosis in subjects having APOEepsilon3epsilon4 genotype. Additional studies in larger patient groups are needed to confirm these observations. PMID- 11388661 TI - Neurotrophic factor therapy--prospects and problems. AB - Over the past 15 years neurotrophic factors have generated considerable excitement for their potential as therapy for a wide variety of degenerative neurological disorders, for which there is currently no treatment. The first part of this period was marked by the discovery, characterization, and cloning of many new growth factors, and by successful testing of these factors in animal models of neurological disease. In recent years the biotechnology industry and pharmaceutical industry have attempted to replicate the success of the animal studies in clinical trials. Although some studies have demonstrated moderate efficacy, for the most part the clinical trials have been less successful at demonstrating the therapeutic efficacy of this new class of drugs. For example, nerve growth factor appeared to be efficacious in two phase II clinical trials for peripheral neuropathy, but failed in a large scale phase III trial. Ciliary neurotrophic factor, brain derived neurotrophic factor and insulin like growth factor-1 have all been tested in clinical trials for the treatment of amyotrophic lateral sclerosis, with at best, variable indications of efficacy. Nevertheless, there are still many reasons to be optimistic that some of these agents may be useful clinically. Many technical and pharmacological issues remain to be adequately addressed, before neurotrophic factors can live up to their potential. Our collective experience with them has re-adjusted previously wild expectations, so that they are now much more realistic. This is necessary and beneficial for the maturation of this field of study. PMID- 11388662 TI - Cell therapy and transplantation in Parkinson's disease. AB - Transplanted human fetal dopamine neurons can reinnervate the striatum in patients with Parkinson's disease (PD). Recent findings using positron emission tomography indicate that the grafts are functionally integrated and restore dopamine release in the patient's striatum. The grafts can exhibit long-term survival without immunological rejection and despite an ongoing disease process and continuous antiparkinsonian drug treatment. In the most successful cases, patients have been able to withdraw L-dopa treatment after transplantation and resume an independent life. About two-thirds of grafted patients have shown clinically useful, partial recovery of motor function. The major obstacle for the further development of this cell replacement strategy is that large amounts of human fetal mesencephalic tissue are needed for therapeutic effects. Stem cells hold promise as a virtually unlimited source of self-renewing progenitors for transplantation. The possibility to generate dopamine neurons from such cells is now being explored using different approaches. However, so far the generated neurons have survived poorly after transplantation in animals. PMID- 11388663 TI - Matrix metalloproteinases: potential therapeutic target in spinal cord injury. AB - Mediators of extracellular matrix proteins degradation, the matrix metalloproteinases (MMPs), involved in inflammation as well as facilitation of process outgrowth of oligodendrocytes are interesting targets for neural repair. Recent data reported their activation after seizures, cerebral ischemia and spinal cord injury. The present study was designed to localize at cellular level the gelatinase activity by in situ zymography in a rat spinal cord contusion model. The kinetic of gelatinase activation was monitored by in situ zymography on 20 microm cryostat sections. The fluorescein-quenched DQ gelatin digestion yielded cleaved fluorescent peptides enabling the detection of gelatinase activity at cellular level. Twenty four hours and 48 h after injury, a strong gelatinase activity was detected at the lesion site in and around vascular structures and infiltrated cells. A preincubation with either MMP-2 or MMP-9 antibodies significantly decreases the gelatinase activity pattern, suggesting the involvement of at least both MMPs. Our results are consistent with a role for MMPs in the blood spinal barrier disruption, the leukocytes infiltration, the disruption of the extracellular matrix and the clearance of debris. PMID- 11388664 TI - Disparate cleavage of poly-(ADP-ribose)-polymerase (PARP) and a synthetic tetrapeptide, DEVD, by apoptotic cells. AB - In the present investigations, we have shown differential cleavage of cellular PARP and a caspase 3-selective synthetic tetrapeptide substrate, Z-DEVD-AFC or Ac DEVD-AMC using a T lymphoblastoid cell line Jurkat, and its variant clone E6.1(J E6). Anti-Fas antibody-mediated apoptosis resulted in DNA fragmentation and PARP cleavage in both Jurkat and J-E6 cells. However, unlike Jurkat, J-E6 cells did not cleave a synthetic tetrapeptide substrate efficiently. The failure to cleave the DEVD tetrapeptide by apoptotic J-E6 cells was not due to insufficient expression or processing of caspase 3 in J-E6 cells. Interestingly, when the J-E6 cells were transiently transfected with a cDNA encoding caspase 3, efficient cleavage of Z-DEVD-AFC was achieved. The observations that apoptotic J-E6 cells barely cleaved a synthetic DEVD tetrapeptide, but efficiently cleaved endogenous PARP, potentially at the most preferred DEVD site, suggest that active caspases may have disparate characteristics to recognize substrates presented in different context. PMID- 11388666 TI - E2F-1 induced apoptosis. AB - Members of the E2F family of transcription factors play an important role in regulating the cell cycle, and their activity is often perturbed during the development of human malignancies. More recent work has shown that E2F-1 regulates apoptosis as well as proliferation, in part by stabilizing the p53 tumor suppressor, an important mediator of apoptosis. This has led to the suggestion that E2F-1 may function as a tumor surveillance mechanism, detecting aberrant proliferation and engaging apoptotic pathways to protect the organism from developing tumors. PMID- 11388665 TI - A decrease in intracellular zinc level precedes the detection of early indicators of apoptosis in HL-60 cells. AB - Low extracellular zinc concentrations have been associated with the induction of apoptosis. To assess the relationship between intracellular zinc concentration and the rate of apoptosis, cells were grown in media containing 0.5, 25, or 50 microM zinc and analyzed by flow cytometry or fluorescence microscopy. Cells grown in 0.5 microM zinc medium over 48 h showed a successive decrease in intracellular zinc concentration measured by the zinc-specific fluorophore, zinquin. After 18 h in 0.5 microM zinc medium, rhodamine 123 retention decreased. However, the addition of 10 microM zinc to the 0.5 microM medium before 16 h in culture restored rhodamine retention in the cells. After 30 h there was an increase in the number of cells cultured in 0.5 microM zinc medium that bound annexin V-FITC. These data indicated that decreased intracellular zinc concentration preceded early markers of apoptosis, with alterations in mitochondrial transmembrane potential preceding the loss of polarity in the cell membrane. PMID- 11388667 TI - Anti-apoptotic action of macrophage stimulating protein (MSP). AB - MSP is a serum protein belonging to the plasminogen-related kringle domain protein family. In addition to macrophages, epithelial cells are also MSP targets. MSP is a multifunctional factor regulating cell adhesion and motility, growth and survival. MSP mediates its biological activities by activating a transmembrane receptor tyrosine kinase called RON in humans or SKT in mice. MSP can protect epithelial cells from apoptosis by activating two independent signals in the PI3-K/AKT or the MAPK pathway. The MAPK pathway mediates the MSP antiapoptotic effect only if additional signaling pathways are activated through adhesion. This indicates that MSP receptors and integrins, the receptors mediating cell-matrix-dependent adhesion, can collaborate in promotion of cell survival. This adhesion-dependent pathway, which is essential for the MAPK mediated anti-apoptotic effect, remains to be identified. A hypothesis that Stat3 might represent a key component of the adhesion-induced anti-apoptotic pathway is presented in this review. PMID- 11388668 TI - The potential of TRAIL for cancer chemotherapy. AB - Innate and acquired resistance to chemotherapy and radiation therapy has been a major obstacle for clinical oncology. One potential adjunct to such conventional treatments is direct induction of cell death by activation of death receptor mediated apoptosis. TRAIL (tumor necrosis factor (TNF)-related apoptosis inducing ligand), a recently identified member of the growing TNF superfamily, binds to its cognate "death" receptors DR4 and DR5 as well as "decoy" receptors DcR1 and DcR2. Upon binding, rapid apoptosis is enacted in a variety of human cancer cell lines independent of p53 status, but not in normal cell lines. TRAIL treatment results in significant growth suppression of TRAIL-sensitive human cancer xenografts in mice. Furthermore, combination treatment of TRAIL with genotoxic chemotherapeutic agents synergistically suppresses growth of tumor xenografts which are otherwise resistant to treatment with TRAIL or chemotherapy alone. Unlike the other death ligands TNF-alpha or FasL, systemic administration of soluble human TRAIL does not cause toxicity in mice and non-human primates. While further studies are needed to evaluate the possible cytotoxicity of TRAIL especially for human hepatocytes, indications are increasing that TRAIL may be a novel therapeutic agent for human cancer. PMID- 11388669 TI - IGFBP-3 and apoptosis--a license to kill? AB - Insulin-like growth factor binding protein (IGFBP)-3, the major carrier of insulin-like growth factors (IGFs) in the circulation, was first isolated and characterised over a decade ago. More recently, IGFBP-3 has been assigned a role as a putative death-promoting factor, a function that appears, under certain circumstances, to be independent of its IGF-binding ability. This review examines the current evidence for a pro-apoptotic function for IGFBP-3 and speculates on its physiological significance. PMID- 11388670 TI - Induction of apoptosis by the marine sponge (Mycale) metabolites, mycalamide A and pateamine. AB - The marine sponge metabolites mycalamide A (mycalamide) and pateamine are extremely cytotoxic. While mycalamide has been shown to inhibit protein synthesis, the mechanism by which these compounds induce cell death is unknown. Using DNA laddering, Annexin-V staining, and morphological analysis, we demonstrate that both metabolites induce apoptosis in several different cell lines. Furthermore, both mycalamide and pateamine were more potent inducers of apoptosis in the 32D myeloid cell line after transformation with either the ras or bcr-abl oncogenes. This increased sensitivity was also observed in response to the protein synthesis inhibitors cycloheximide and puromycin, and cytosine-beta-D arabinofuranoside (Ara-C), an inducer of DNA damage. We propose, therefore, that in 32D cells where Ras signalling has been altered either by constitutive expression of oncogenic ras or by Bcr/abl-mediated perturbation of upstream signalling events, increased susceptibility to apoptosis by a range of stimuli is conferred. PMID- 11388672 TI - Evaluation of occult acetaminophen hepatotoxicity in hospitalized children receiving acetaminophen. Pediatric Pharmacology Research Unit Network. AB - The safety of repeated doses of acetaminophen in ill children with the potential of reduced glutathione stores has been questioned. This study measured hepatic transaminases in children and adolescents (n=100) who received > or = 6 therapeutic doses of acetaminophen over a 48-hour period of hospitalization. Acetaminophen-protein adducts were measured in a cohort of subjects with hepatic transaminase elevation (n=8) and in those (n=10) receiving concurrent drug therapy with agents that induce the cytochrome P450 enzymes involved in acetaminophen metabolism. Acetaminophen-protein adducts were not detected in this cohort of 18 subjects. Based on this pilot study, the routine use of acetaminophen at therapeutic doses in ill, hospitalized children and adolescents appears safe. PMID- 11388671 TI - A putative protein inhibitor of activated STAT (PIASy) interacts with p53 and inhibits p53-mediated transactivation but not apoptosis. AB - The p53 protein has recently been reported to be capable of mediating apoptosis through a pathway that is not dependent on its transactivation function. We report here that the PIASy member of the protein inhibitor of activated STAT family inhibited p53's transactivation function without compromising its ability to induce apoptosis of the H1299 nonsmall cell lung carcinoma cell line. The p53 protein bound to PIASy in yeast two-hybrid assays and coprecipitated in complexes with p53 in immunoprecipitates from mammalian cells. PIASy inhibited the DNA binding activity of p53 in nuclear extracts and blocked the ability of p53 to induce expression of two of its target genes, Bax and p21Waf1/Cip1, in H1299 cells. The block in p53-mediated induction of Bax and p21 was determined to be at the level of transactivation, since PIASy inhibited p53's ability to transactivate a p21/luciferase reporter construct. PIASy did not effect the incidence of apoptosis in H1299 cells upregulated for p53. PIASy appears to regulate p53-mediated functions and may direct p53 into a transactivation independent mode of apoptosis. PMID- 11388673 TI - Frequency of celiac disease in individuals with Down syndrome in the United States. AB - Ninety-three individuals with Down syndrome (DS) were screened to investigate the prevalence of celiac disease (CD) in the United States. Five of the 93 individuals were antiendomysial antibody (EMA) positive. Of the 5 who tested positive for EMA, 4 were biopsied, 1 refused biopsy. Three of the 4 individuals biopsied manifested changes of CD on small bowel biopsy. This gives a frequency of 3.2% of confirmed CD in our DS individuals and suggests the need for periodic screening for celiac disease in this population. PMID- 11388674 TI - Intervention strategies for children poorly adherent with asthma medications; one center's experience. AB - Poor adherence to medications and other aspects of the treatment plan is common in pediatric patients with asthma, and is a common reason for inadequate asthma control. In selected patients we have used electronic monitoring of pulmonary function, behavior contracts, home nursing visits, and medical neglect reports in an attempt to improve adherence and asthma control. Improved outcomes were seen with the most aggressive intervention, home nursing, and medical neglect referral, but not with less aggressive measures. PMID- 11388675 TI - A program description of health care interventions for homeless teenagers. AB - This prospective review was designed to determine the effectiveness of a broad spectrum health intervention program for homeless and runaway youth. Diagnosis, treatment, and counseling for drug use, sexually transmitted diseases (STDs), and other health issues were provided all new admissions to a residential care facility during a 2-month enrollment. Education was continued during a 9-month follow-up period based on the program entitled Bright Futures, previously developed and published by the National Center for Education in Maternal and Child Health. Sixty percent of the 106 study residents had STDs on admission and 7% developed new STDs after completing therapy and undergoing counseling. Drug dependence was reduced from 41% to 3%, and 42% achieved full-time or part-time employment. Fifty-nine percent completed hepatitis B immunization with the 3-dose series. This experience suggests that an organized program of interventions in a residential care facility for homeless teenagers can significantly reduce drug dependence and STDs. PMID- 11388676 TI - The use of dietary supplements in pediatrics: a study of echinacea. AB - Alternative medical therapies are commonly used and have increased in popularity. Although patients may not always disclose the use of alternative therapies, they may seek advice regarding their use, especially for children. Regulation and standardization of these modalities, especially botanicals, is incomplete. The University of Arizona has initiated a study of the use of echinacea in the prevention of recurrent otitis media. A review of echinacea preparations was undertaken, and this report discusses the complexities surrounding the use of this dietary supplement. The number and diversity of echinacea preparations are detailed; the role of the physician as "botanical" advisor to patients and families is examined. PMID- 11388677 TI - Herbal medicine. PMID- 11388678 TI - Recurrent encephalopathy and seizures in an adolescent girl. PMID- 11388679 TI - Acute myocardial ischemia in a healthy male child: an atypical presentation of acute Epstein-Barr virus infection. PMID- 11388680 TI - Atypical seizures as the cause of apnea in a 6-month-old child. PMID- 11388681 TI - Recurrent pulmonary edema: an uncommon presenting feature of childhood obstructive sleep apnea hypoventilation syndrome in an otherwise healthy child. PMID- 11388682 TI - The effect of fiber supplementation on lipid profile in children with hypercholesterolemia. PMID- 11388683 TI - Is infantile colic a migraine-related phenomenon? PMID- 11388685 TI - Hypereosinophilia and streptococcal infection. PMID- 11388684 TI - Scarring is the central issue in urinary tract infection, not vesicoureteral reflux. PMID- 11388686 TI - Electrophysiology of growth control and acupuncture. AB - Bioelectric fields have been shown to interact with morphogens and guide growth control. The morphogenetic singularity theory published a decade ago suggests that organizing centers have high density of gap junctions and high electrical conductance. They are the singular points in morphogen gradient and bioelectric field. A growth control system originates from a network of organizing centers containing under-differentiated cells and retains its regulatory functions after embryogenesis. The formation and maintenance of all the physiological systems are directly dependent on the activity of the growth control system. The evolutionary origin of the growth control system is likely to have preceded all the other physiological systems. Its genetic blueprint might have served as a template from which the newer systems evolved. The growth control signal transduction is embedded in the activity of the function-based physiological systems. The regulation of most physiological processes is through growth control mechanisms such as hypertrophy, hyperplasia, atrophy, and apoptosis. Acupuncture points, which also have high electrical conductance and high density of gap junctions, originate from organizing centers. This theory can explain the distribution and non-specific activation of organizing centers and many research results in acupuncture. In several 'prospective blind trials', recent research results have supported its corollary on the role of singularity and separatrix in morphogenesis, the predictions on the high electric conductance and the high density of gap junctions at the organizing centers. These advances have broad implications in biomedical sciences. PMID- 11388687 TI - Adenosine enhances glial glutamate efflux via A2a adenosine receptors. AB - The present study investigated the effect of adenosine on glial glutamate efflux. Adenosine (from 1 nM to 100 microM) enhanced the release from cultured rat glial cells in a bell-shaped dose-responsive manner for the hippocampus and in a dose dependent manner for the superior colliculus, and a similar increase was obtained with the A2a adenosine receptor agonist, 2-p-(2-carboxyethyl) phenethylamino-5'-N ethylcarboxamidoadenosine hydrochloride (CGS21680), but not with the A1 adenosine receptor agonist, N6-cyclohexyladenosine (CHA). Adenosine and CGS21680 also enhanced glutamate efflux from Xenopus oocytes injected with the poly (A)+ mRNAs derived from cultured glial cells for the hippocampus and the superior colliculus together with and without the A2a adenosine receptor mRNA, but instead such increase was not found in oocytes expressing A2a adenosine receptors alone. The results of the present study thus suggest that adenosine enhances glutamate efflux from glial cells via A2a adenosine receptors, and this may represent a mechanism underlying the facilitatory action of adenosine on hippocampal and superior colliculus neurotransmissions. PMID- 11388688 TI - In vitro evaluation of the antioxidant activity and biomembrane interaction of the lazaroid U-74389G. AB - The aim of this paper was to clarify whether the interaction of the lazaroid U 74389G with phospholipid membranes might be relevant as to its antioxidant activity. Thus we evaluated the "in vitro" antioxidant activity of U-74389G in two experimental models: 1) bleaching of the stable 1,1-diphenyl-2-picrylhydrazyl radical; 2) peroxidation, induced by the water-soluble radical initiator 2,2' azobis(2-amidino-propane) hydrochloride, on mixed dipalmitoyl phosphatidylcholine/linoleic acid unilamellar vesicles. Moreover, given that biophysical techniques may help in explaining the role of a drug in its interaction with the microenvironment of the model lipid membranes, we used a classical approach to investigate the U-74389G/model membrane interaction: the differential scanning calorimetry technique on dimyristoylphosphatidylcholine multilamellar and unilamellar vesicles and the Langmuir-Blodgett technique on dimyristoylphosphatidylcholine monolayers. The results evidenced the strong antioxidant activity of U-74389G (especially in a membranous system) and its capability to interact with and be transported across model membranes. Thus one can speculate that U-74389G can act as scavenger of chain-propagating lipid peroxyl radicals within the membranes and may be able to protect not only cell membranes, but also intracellular components against peroxidative attack. Furthermore, also if there is no certain proof that the effect on the lipid packing order may play a key role in its antioxidant activity, the fluidifying effect on phospholipid bilayers of U-74389G favourably complements its free radical scavenging characteristics. PMID- 11388689 TI - Prior exposure to methylenedioxyamphetamine (MDA) induces serotonergic loss and changes in spontaneous exploratory and amphetamine-induced behaviors in rats. AB - The substituted amphetamine 3,4 methylenedioxyamphetamine (MDA) is a popular recreational drug of abuse. Administration of MDA to experimental animals has been shown to induce damage to serotonergic axons and nerve terminals. However, there is a lack of information on whether these treatments can produce any long term changes in behavioural performance particularly under stressful conditions. In the present study, MDA (7.5 mg/kg; i.p.) was administered twice daily to adult male Sprague Dawley rats for four days. Four weeks following the last dose, spontaneous behaviors of these animals were tracked and scored in a novel "open field" environment using an automated video registration and computer interpretation system. Changes in behavior were observed in MDA treated animals including reductions in exploratory oriented behaviors (locomotion and rearing) and increases in grooming behavior when compared to vehicle treated controls. MDA treated animals also displayed an enhanced locomotor and stereotyped response to d-amphetamine (12 mg/kg; i.p.). Significant reductions in 5-HT concentrations (20 30%) were observed in the frontal cortex, amygdala, striatum, and hypothalamus as a result of MDA treatment. In addition, [3H] paroxetine binding was reduced by (40%) in the cortex of MDA treated rats indicating that the decrease in 5-HT concentrations were accompanied by a reduction in intact presynaptic 5-HT nerve terminals. Changes in behavioural performance in a novel "open field" environment and following d-amphetamine challenge might be considered as a behavioural model of serotonergic deficit induced by MDA. The findings of this study also suggest that MDA use may increase both the abuse potential, and the propensity to develop psychosis as a result of abusing other psychostimulants such as d-amphetamine. PMID- 11388690 TI - Modulation of diaspirin crosslinked hemoglobin induced systemic and regional hemodynamic response by ethanol in normal rats. AB - DCLHb, a hemoglobin based oxygen carrier, has been extensively studied for the treatment of hemorrhagic shock in both animal models and humans. Numerous accidents resulting in trauma are due to ethanol intoxication, in particular cases of car accidents. Therefore, trauma patients might be intoxicated with drugs of abuse like ethanol. Ethanol has significant effects on the cardiovascular system including peripheral vasodilation and decreased myocardial contractility. Such effects are likely to alter the cardiovascular actions of DCLHb, a resuscitative agent. Hence, this study investigated the effect of ethanol on the cardiovascular actions of DCLHb. Urethane anesthetized male Sprague-Dawley rats were divided into following groups (i) Saline + DCLHb (400 mg/kg) (n = 9), (ii) Ethanol (1 g/kg) + DCLHb (400 mg/kg) (n = 9), and (iii) Ethanol (4 g/kg) + DCLHb (400 mg/kg) (n = 8). Cardiovascular parameters were determined using a radioactive microsphere technique. DCLHb when administered to saline treated rats produced an increase in MAP, TPR, decreased renal and hepatic blood flow and increased blood flow to the skin and mesentery & pancreas. A high dose of ethanol (4 g/kg) significantly attenuated the DCLHb induced pressor response (p < 0.05) and increase in TPR (p < 0.05). Cardiac output was severely reduced by DCLHb in rats treated with high dose ethanol as compared to saline. No changes in TPR and cardiac output were observed in the low dose ethanol (1 g/kg) group. DCLHb reduced blood flow to the heart and mesentery & pancreas in rats treated with high dose ethanol. DCLHb caused a decrease in musculo-skeletal vascular resistance in rats treated with high dose ethanol. This decrease in vascular resistance in the musculo-skeletal system appears to be contributing to a decrease in TPR. It is concluded that ethanol in higher doses significantly alters the hemodynamic effects of DCLHb and may interfere with the resuscitative effects of DCLHb. PMID- 11388691 TI - Opposing pharmacological actions of cepharanthin on lipopolysaccharide-induced histidine decarboxylase activity in mice spleens. AB - A biscoclaurin alkaloid preparation, cepharanthin (Ceph), is reported to have opposing pharmacological effects, enhancement or depression, on several cells and tissues, although detailed mechanisms remain unclear. Previously, we reported that Ceph enhanced lipopolysaccharide (LPS)-induced histidine decarboxylase (HDC) activity in mice spleens by consecutive pre-administration. In this study, we examined the pharmacological effects on HDC activity of a single Ceph pre administration to test the influence of the administration method. Consequently, HDC activities were decreased by a single administration 15 minutes before LPS challenge in ddY and ICR mice spleens. Moreover, to further examine this suppressing effect, we employed genetically mast cell-deficient WBB6F1 W/Wv (W/Wv) mice to avoid the influence of mast cells. In W/Wv mice, HDC activity was enhanced, but not in the congenic WBB6F1 +/+ mice. These findings suggest that mast cells influence the depressant effect on HDC activity by a Ceph single administration in mast cell sufficient mice. PMID- 11388692 TI - Mechanisms involved in gastric protection of melatonin against oxidant stress by ischemia-reperfusion in rats. AB - The generation of oxygen-derived free radicals has been suggested to be significantly responsible for ischemia-reperfusion injury in gastrointestinal tissues. Biochemical mechanisms include the xanthine-oxidase-derived oxidants mainly the superoxide anion. Both in vitro and in vivo studies have demonstrated that the pineal hormone melatonin possesses free radical scavenging and antioxidant properties. The indolamine has been effective in reducing the induced oxidative damage in several tissues and biological systems. The aim of this study was to elucidate additional antioxidant mechanisms responsible for the gastroprotection afforded by the indolamine in ischemia-reperfusion gastric injury. Therefore, changes of related enzymes such as xanthine-oxidase, superoxide dismutase, glutathione reductase and total glutathione were investigated. Our results showed that treatment with 5, 10 or 20 mg kg(-1) of melatonin, administered i.p., clearly diminished the percentage of damage to 49.56 +/- 17.20, 37.54 +/- 11.40 and 26.70 +/- 8.12 respectively. Histologically there was a reduction of exfoliation of superficial cells and blood cell infiltration. These protective effects were related to a significant reduction of xanthine-oxidase activity (2.23 +/- 0.38 U/mg prot x 10(-4) with the highest tested dose of melatonin) and significant increases in superoxide dismutase reaching a value of 6.20 +/- 0.56 U/mg prot with 25 mg/Kg of melatonin and glutation reductase activities (417.44 +/- 29.72 and 649.43 +/- 81.11 nmol/min/mg prot with 10 and 20 mg/Kg of melatonin). We conclude that the free radical scavenger properties of melatonin mainly of the superoxide anion, probably derived via the xanthine-oxidase pathway, and the increase of antioxidative enzymes significantly contributes to mediating the protection by the hormone against ischemia-reperfusion gastric injury. PMID- 11388693 TI - Insulin hyperresponsiveness in partially hepatectomized diabetic rats. AB - The present work analyzes the expression of insulin receptors and theirs related intracellular signaling molecules in partially hepatectomized-diabetic rats. Insulin binding through Scatchard analysis was studied using isolated hepatocytes of Control (Sham-operated), Hepatectomized, Diabetic and Diabetic-Hepatectomized male Wistar rats. In a set of in vivo experiments, the levels of alpha subunit of the insulin receptor, the insulin receptor substrate 1 (IRS-1) and the phosphatidylinositol 3-kinase (PI3K) were determined. [3H]-thymidine incorporation into DNA 24 or 48 h after surgery was assessed in all the experimental groups. Scatchard analysis showed that insulin receptor number was increased in diabetic and in hepatectomized rats in the same extent (64%, with respect to Controls). Diabetic-hepatectomized rats showed a dramatic increase of the receptor concentration (400%) and on the affinity constant (532%). Besides, the insulin receptor expression was increased in the treated groups, being the higher values those of the diabetic-hepatectomized rats. IRS-1 and PI3K showed similar increases. DNA synthesis was not impaired by the diabetes state. In conclusion, increased expression of IR and IRS-1 leads to increased association of PI3K in vivo in diabetic regenerating rats. The enhancement of this pathway may reveal an insulin hyperresponsiveness in these animals. PMID- 11388694 TI - Evidence for expression of estrogen receptor cofactor messenger ribonucleic acid in the ovary and uterus of domesticated animals (sheep, cow and pig). AB - Expression levels of estrogen receptor cofactors (coactivators or corepressors) in specific tissue compartments and cells are thought to influence the expression of estrogen responsive genes and thereby influence overall hormonal responsiveness of target tissues. To date, the presence of cofactors has been reported in tissues from humans, rats and mice. We analyzed the presence and distribution of messenger ribonucleic acids (mRNAs) encoding several transcriptional cofactors in the ovary and uterus of three domestic animal species, the sheep, cow and pig. Northern analysis for cofactors SRC-1, GRIP1, RAC3, p300, RIP140, and SPA showed expression in ovaries from all three species. In addition, lower expression of SRC-1, GRIP1, RAC3, p300, and RIP140 mRNAs was observed during the luteal phase (day 10-12 of the estrous cycle) than at estrus (day 0); however, SPA transcript levels remained unchanged. We then examined expression of mRNAs for changing (SRC-1, RIP140) and constitutively expressed (SPA) cofactors in ovine ovaries. SRC-1 and RIP140 transcripts in corpus luteum were lower compared to the surrounding ovarian tissue. SPA mRNA expression, however, was similar in corpus luteum and surrounding tissues. To determine which ovarian cell types express SRC-1, RIP140, and SPA, in situ hybridization was performed on sheep ovaries. Silver grains corresponding to these cofactors were seen in ovarian granulosa, theca and stromal cells, but appeared to be most abundant in the granulosa cells. Expression of SRC-1 and RIP140 in corpus luteum, however, was reduced compared to expression in follicular cells. Finally, we examined cofactor expression in ovine, bovine, and porcine uterus. Northern blot analysis for SRC-1, GRIP1, RAC3, p300, and RIP140 mRNAs showed higher expression in extracts of the endometrium compared to whole uterus. We provide the first evidence for the presence of estrogen receptor cofactor mRNAs in the ovary and uterus of three domestic animal species. We suggest that coactivators are conserved among species and associated with hormonal responsiveness of reproductive tract tissues in sheep, cow and pig. PMID- 11388695 TI - The effects of zenarestat, an aldose reductase inhibitor, on minimal F-wave latency and nerve blood flow in streptozotocin-induced diabetic rats. AB - The effects of zenarestat, 3-(4-bromo-2-fluorobenzyl)-7-chloro-3,4-dihydro-2,4 dioxo-1(2H)-quinazolineacetic acid, an aldose reductase inhibitor (ARI), on F wave conduction abnormalities, nerve blood flow (NBF) reduction and sorbitol accumulation were studied in streptozotocin-induced diabetic rats. Two weeks after the induction of diabetes, zenarestat was given once a day for two weeks. In diabetic control rats, marked accumulation of sorbitol, reduction of NBF and prolongation of minimal F-wave latency (FWL) were observed as compared to normal rats. Zenarestat, at a dose of 32 mg/kg, inhibited sorbitol concentration to nearly the normal rat level and significantly improved not only NBF but also minimal FWL. At a dose of 3.2 mg/kg, sorbitol accumulation was inhibited by approximately 40% and there was a tendency to increase in NBF; however, minimal FWL was not improved at all. These data suggest that a highly inhibition of the nerve sorbitol accumulation is requisite for the treatment of diabetic peripheral neuropathy. PMID- 11388696 TI - Relationship between diurnal rhythm of cell cycle and interferon receptor expression in implanted-tumor cells. AB - Whether the diurnal rhythm of cell cycle is associated with that of interferon alpha/beta receptor (IFNAR) expression was investigated in implanted-tumor cells. The expression of IFNAR mRNA significantly increased when the proportion of tumor cells in DNA synthesis (S) phase increased in vitro. A diurnal rhythm was observed for cell cycle distribution in implanted-tumor cells. The specific binding of interferon-alpha to receptor and IFNAR mRNA increased when the proportion of tumor cells in S phase increased in vivo. The time-dependent expression of IFNAR was supported by that of transcription factor level induced by interferon-beta. The present result suggests that the rhythm of IFNAR expression is closely related to that of cell cycle distribution in implanted tumor cells. PMID- 11388697 TI - Acute alcohol intoxication and endotoxemia desensitize HIV-1 gp120-induced CC chemokine production by Kupffer cells. AB - Chemokines are involved in the inhibition of HIV-1 infection and in the pathogenesis of tissue injury in a number of conditions, including endotoxemia and alcoholic liver disease. CC chemotactic peptides (MIP-1alpha, MCP-1 and RANTES) are produced by a wide variety of cell types in response to immunological stimuli, bacterial endotoxin and gp120 from HIV-1 and HIV-2. This work tests the hypothesis that prior exposure to endotoxin and/or ethanol in vivo inhibits the production of CC-chemokines following a secondary challenge with HIV-1 gp120 in vitro. Male Sprague-Dawley rats received in intravenous infusion of ethanol to maintain blood ethanol level at 170 mg/dl for 3 hr. Escherichia coli LPS (1 mg/Kg) was given intravenously 5 min after the ethanol bolus was injected. Control groups received similar volumes of saline. Three hr after LPS treatment, Kupffer cells were obtained and treated with HIV-1 gp120 (5 microg/10(6) cells/24 hr). At the end of the incubation period, cells were obtained for RT-PCR analysis of CC-chemokine mRNA expression. Chemokine release in culture supernatants was measured by ELISA. Results show that in vivo ethanol was associated with downregulation of MIP-1alpha and MCP-1 mRNA expression and protein release in primary cultures of Kupffer cells. However, ethanol alone primed isolated Kupffer cells for enhanced RANTES mRNA and protein release in the presence or absence of HIV-1 gp120. These results demonstrate that acute ethanol intoxication and endotoxemia may selectively act as a desensitizing agent in response to a secondary challenge with bacterial or viral products. PMID- 11388698 TI - Body temperature dependency in baclofen-induced gastric acid secretion in rats relation to capsaicin-sensitive afferent neurons. AB - Body temperature dependency in gastric functional responses to baclofen, a GABA(B) agonist, such as acid secretion, mucosal blood flow (GMBF) and motor activity, was examined in urethane-anesthetized rats under normal (37+/-1 degrees C) and hypothermic (31+/-1 degrees C) conditions. A rat stomach was mounted in an ex-vivo chamber, perfused with saline, and the acid secretion was measured using a pH-stat method, simultaneously with GMBF by a laser Doppler flowmeter. Gastric motility was measured using a miniature balloon as intraluminal pressure recordings. Intravenous administration of baclofen significantly increased acid secretion at the doses > 0.3 mg/kg under hypothermic conditions, yet it caused a significant stimulation only at doses > 10 mg/kg under normothermic conditions. The increases in gastric motility and GMBF were similarly induced by baclofen, irrespective of whether the animals were subjected to normothermic or hypothermic conditions. These functional responses to baclofen under hypothermic conditions were totally attenuated by either bilateral vagotomy or atropine (3 mg/kg, s.c.). Baclofen at a lower dose (1 mg/kg i.v.) significantly increased the acid secretion even under normothermic conditions when the animals were subjected to chemical deafferenation of capsaicin-sensitive neurons or pretreatment with intracisternal injection of CGRP8-37 (30 ng/rat). These results suggest that 1) gastric effects of baclofen are dependent on body temperature in stimulating acid secretion but not GMBF or motor activity, 2) the acid stimulatory action of baclofen is enhanced under hypothermic conditions, and 3) the suppression of baclofen-induced acid response under normothermic conditions may be related to capsaicin-sensitive afferent neuronal activity, probably mediated by central release PMID- 11388699 TI - Involvement of central GABAergic neurons in basal and diurnal changes of tuberoinfundibular dopaminergic neuronal activity and prolactin secretion. AB - Central administration of gamma-aminobutyric acid (GABA) has been shown to stimulate the secretion of prolactin (PRL). Whether GABA acts via dopamine, the major PRL-inhibiting hormone, and which GABA receptor type(s) is involved have not been ascertained. Both GABA(A) and GABA(B) receptor agonists and/or antagonists were administered centrally in this study and their effects on both basal and diurnal changes of tuberoinfundibular dopaminergic (TIDA) neuronal activity were determined by measuring the concentration of 3,4 dihydroxyphenylacetic acid (DOPAC) in the median eminence (ME). Serum PRL level was determined by RIA. Ovariectomized, estrogen-primed Sprague-Dawley rats implanted with intracerebroventricular (icv) cannulae were used. Muscimol (1 ng/3 microl/rat, icv), a GABA(A) receptor agonist, but not baclofen (1-100 ng/3 microl/rat, icv), a GABA(B) receptor agonist, injected in the morning significantly lowered and elevated ME DOPAC and serum PRL levels, respectively at 15 and 30 min. Lower and higher doses of muscimol were not effective. The effects of muscimol could also be prevented by co-administration of bicuculline (0.1-10 ng/3 microl, icv), a GABA(A) receptor antagonist. When bicuculline (10-500 ng/3 microl, icv) was given in the afternoon (at 1500 h), it significantly reversed the lowered ME DOPAC level in the afternoon and prevented the concurrent PRL surge. We conclude that endogenous GABA acting through GABA(A) receptors may play a significant role in the control of basal and diurnal changes of TIDA neuronal activity, and in turn, PRL secretion. PMID- 11388700 TI - Characterization of human, dog and rabbit corpus cavernosum type 5 phosphodiesterases. AB - Human, dog and rabbit corpus cavernosum type 5 phosphodiesterases (PDE5) were isolated and their characteristics were compared. The three enzymes showed Km values of 0.8, 2.1 and 2.3 uM, respectively. They exhibited similar pH-dependence with optimal pH being 7.5. They required Mg++ for activity and the activity was suppressed by high concentrations of Zn++ (0.1-1 mM). Sildenafil potently inhibited the three enzymes with IC50 values of 3.6, 1.7 and 3.0 nM, respectively. Dipyridamole and IBMX (3-isobutyl-1-methylxanthine) each also inhibited the three enzymes with similar, albeit lower, potencies (IC50 about 1.1 and 5.7 uM, respectively). However, zaprinast exhibited a significantly higher potency against the rabbit enzyme (IC50 53 nM) than against the human and dog PDE5s (IC5s 332 and 217 nM, respectively). Thus, the corpus cavernosum PDE5s are very similar among the various species with the only significant difference being their sensitivity to zaprinast. Human platelet PDE5 was also characterized by comparison with the corpus cavernosum enzyme. The platelet enzyme exhibited a Km, pH-, Mg++- and Zn++-dependence, and sensitivity to sildenafil and zaprinast very similar to those of the corpus cavernosum PDE5. However, compared with corpus cavernosum PDE5, the platelet enzyme exhibited higher sensitivity to dipyridamole and IBMX (IC50 0.46 and 1.8 uM, respectively). This study shows that despite similar kinetics and enzymatic properties, corpus cavernosum PDE5s from different species, and corpus cavernosum and platelet PDE5s, can have differential sensitivity to pharmacological inhibitors. PMID- 11388701 TI - Thrombin activates p38 mitogen-activated protein kinase in vascular smooth muscle cells. AB - Thrombin is a potent mitogen for vascular smooth muscle cells. However, the signaling pathways by which thrombin mediates its mitogenic response are not fully understood. The ERK (extracellular signal-regulated protein kinase) and JNK (c-Jun N-terminal kinase) members of the mitogen-activated protein kinase (MAPK) family are reported to be activated by thrombin. We have investigated the response to thrombin of another member of the MAPK family, p38 MAPK, which has been suggested to be activated by both stress and inflammatory stimuli in vascular smooth muscle cells. We found that thrombin induced time- and dose dependent activation of p38 MAPK. Maximal stimulation of p38 MAPK was observed after a 10-min incubation with 1 unit ml(-1) thrombin. GF109203X, a protein kinase C inhibitor, and prolonged treatment with phorbol 12-myristate 13-acetate partially inhibited p38 MAPK activation. A tyrosine kinase inhibitor, genistein, also inhibited p38 MAPK activation in a dose-dependent manner. p38 MAPK activation was inhibited by overexpression of betaARK1ct (beta-adrenergic receptor kinase I C-terminal peptide). p38 MAPK activation was also inhibited by expression of dominant-negative Ras, not by dominant-negative Rac. We next examined the effect of a p38 MAPK inhibitor, SB203580, on thrombin-induced proliferation. SB203580 inhibited thrombin-induced DNA synthesis in a dose dependent manner. These results suggest that thrombin activates p38 MAPK in a manner dependent on Gbetagamma, protein kinase C, a tyrosine kinase, and Ras, that p38 MAPK has a role in thrombin-induced mitogenic response in the cells. PMID- 11388702 TI - Mouse erythrocytes as carriers for coencapsulated alcohol and aldehyde dehydrogenase obtained by electroporation in vivo survival rate in circulation, organ distribution and ethanol degradation. AB - Alcohol dehydrogenase and aldehyde dehydrogenase (ADH and ALDH) have been coencapsulated into mouse erythrocytes by an electroporation technique. The optimal conditions were achieved as follows: 420 V, four pulses of 1 ms every 15 min. at 37 degrees C, completed by resealing: 1 h at 37 degrees C. An encapsulation yield ranging from 11-12% was obtained for ADH+ALDH-loaded erythrocytes. Carrier cell recovery was 52%. Electroporated-RBCs observed under Scanning electron microscopy exhibited a tendency toward invaginated sphero stomatocytes. These invaginations were not found in electroporated/resealed RBCs. The intravenous administration of 51Cr-RBCs manifested a bimodal pharmacokinetic profile: an initial phase (t1/2alpha) with a rapid decrease of plasma 51Cr-RBCs followed by a slow and prolonged elimination phase (t1/2beta). The values corresponding to in vivo survival rate during the elimination phase indicated that the survival rate of 51Cr-electroporated loaded-RBCs was slightly lower (t1/2beta, 4.5 days) than 51Cr-native RBCs (t1/2beta, 5.3 days). The mean clearance values from blood of electroporated 51Cr-RBCs (unloaded and loaded) were higher (0.51 %51Cr/day and 0.54 %51Cr/day, respectively) than the obtained for native 51Cr-RBCs (0.18 %51Cr/day). The target organs for electroporated RBCs proved to be the same as for native RBCs. However, electroporated RBCs showed highest accumulation in liver, spleen and lung, since they were promptly recognized by the reticuloendothelium system. Mice induced to the state of acute ethanol intoxication and treated with ADH+ALDH-RBCs clearly showed a lower level of ethanol concentration in plasma (less than 43% ethanol) than the intoxicated mice treated with native RBCs. En consequence the clearance values of ethanol from blood in intoxicated mice treated with ADH+ALDH-RBCs (0.39 ml/min) were higher than the treated with native RBCs (0.20 ml/min). The results obtained suggest that ADH+ALDH-loaded erythrocytes could be used as a potential carrier system for in vivo removal of high levels of ethanol from blood caused by excessive alcohol consumption. PMID- 11388703 TI - Inhibition by a novel anti-arrhythmic agent, NIP-142, of cloned human cardiac K+ channel Kv1.5 current. AB - NIP-142 was shown to prolong atrial effective refractory period and to terminate atrial fibrillation and flutter in in vivo canine models. To obtain information on its antiarrhythmic action, we examined the effect of NIP-142 on cloned human cardiac K+ channel Kv1.5 (hKv1.5) currents stably expressed in a human cell line using whole-cell voltage clamp methods. NIP-142 inhibited the hKv1.5 current in a concentration-dependent and voltage-independent manner. The inhibition was larger at the end of depolarizing pulse than at the outward current peak. The IC50 for inhibition of the steady-state phase was 4.75 microM. A cross-over phenomenon was observed when current traces in the absence and presence of NIP-142 were superimposed. Inhibition of hKv1.5 current by NIP-142 was frequency-independent; changing the depolarizing pulse frequencies (0.1, 0.2, 1 Hz) and little effect on the degree of inhibition. NIP-142 decreased the maximal peak amplitude of kHv1.5 current at the first command pulse after 3 min rest in the presence of the drug. These results suggest that NIP-142 has inhibitory effects on the hKv 1.5 current through interaction with both open and closed states of the channel, which may underlie its antiarrhythmic activity in the atria. PMID- 11388704 TI - Prodigiosin-induced apoptosis in human colon cancer cells. AB - Prodigiosin is a red pigment produced by various bacteria including Serratia marcescens. Colorectal cancer is one of the most frequent malignancies and one of the most frequent causes of cancer death in the Western world. Its treatment is far from satisfactory and the challenge to oncologists is to find novel chemical entities with less toxicity and greater effectiveness than those used in current chemotherapy. Here we characterize the apoptotic action of prodigiosin in colon cancer cells. DLD-1 and SW-620 human colon adenocarcinoma cells, NRK and Swiss 3T3 nonmalignant cells were assayed by the MTT assay, fragmentation pattern of DNA, Hoechst 33342 staining and study of PARP cleavage by Western blot, in order to characterize the prodigiosin-induced apoptosis. Prodigiosin was purified and its structure was confirmed. Metastatic SW-620 cells were more sensitive to prodigiosin (IC50: 275 nM) than DLD-1. We did not observe a significant decrease in the viability of NRK cells. We confirmed that prodigiosin induces apoptosis in both cancer cell lines by the characteristic DNA laddering pattern and condensed nuclei or apoptotic bodies identified by fluorescence microscopy. These results indicate that prodigiosin induces apoptosis in colon cancer cells. PMID- 11388705 TI - Antioxidant status and lipoprotein peroxidation in chronic fatigue syndrome. AB - The aetiology and pathogenesis of the Chronic Fatigue Syndrome (CFS) are still largely unresolved. Accompanying metabolic disorders such as selective n-6 fatty acid depletion suggest that oxidative stress and more specifically lipid peroxidation might play a role in its pathogenesis. In order to investigate this hypothesis, oxidant-antioxidant status and its impact on lipoprotein peroxidation in vitro was examined in 61 patients with unexplained fatigue lasting more than 1 month. They were subdivided into 2 groups: group CFS+ (33 subjects) fulfilled the 1988 Center of Disease Control criteria for CFS and group CFS- did not but was similar as regards age, sex distribution and clinical characteristics. Antioxidant status was similar in the 2 groups except for lower serum transferrin in the CFS + (mean (95 % CI) 2.41 (2.28-2.54) versus 2.73 (2.54-2.92) g/L in the CFS-, p = 0.009) and higher lipoprotein peroxidation in vitro: 6630 (5949-7312) versus 5581 (4852-6310) nmol MDA/mg LDL and VLDL cholesterol x minutes, p = 0.035). CFS intensified the influence of LDL cholesterol (p = 0.012) and of transferrin (p = 0.045) on peroxidation in vitro, suggesting additional pro oxidant effects. These results indicate that patients with CFS have increased susceptibility of LDL and VLDL to copper-induced peroxidation and that this is related both to their lower levels of serum transferrin and to other unidentified pro-oxidising effects of CFS. PMID- 11388706 TI - Rapid down regulation of pyroglutamyl peptidase II activity by arachidonic acid in primary cultures of adenohypophyseal cells. AB - Thyrotropin releasing hormone (TRH; pglu-his-proNH2) is inactivated, in the extracellular space, by pyroglutamyl aminopeptidase II (PPII), a narrow specificity ectopeptidase. In adenohypophysis, multiple hormones regulate PPII surface activity. The intracellular pathways of regulation are still poorly understood. Since some of the neurohormones which regulate PPII activity, including TRH and dopamine, transduce in part their effect through modulation of arachidonic acid (AA) mobilization, we have tested its role in regulation of PPII activity in primary cultures of rat adenohypophyseal cells. Melittin concentrations from 0.25 to 1 ug/ml induced a rapid decrease of PPII activity; 0.5 ug/ml caused a maximum effect (38-45% inhibition) at 20-30 min. AA (0.5 or 5 uM) also inhibited PPII activity (42-72%, maximum at 20 min); AA effect was reversible, with values approaching control at 1 h. The inhibitory effect of AA was blocked by lipoxygenase (10 uM nordihidroguaiaretic acid) but not ciclooxygenase inhibitors (10 uM indomethacin) suggesting the involvement of the lipoxygenase pathway. These data show that production of arachidonic acid by adenohypophyseal cells can rapidly but transiently down regulate surface PPII activity. This is the first evidence that AA mobilization can regulate the activity of an ectopeptidase. PMID- 11388707 TI - Absolute vs. relative numbers in evaluating drug therapy. PMID- 11388708 TI - Vaccine policy decisions: tension between science, cost-effectiveness and consensus? PMID- 11388709 TI - Preemptive analgesia: decreasing pain before it starts. PMID- 11388710 TI - Fingertip injuries. AB - The family physician often provides the first and only medical intervention for fingertip injuries. Proper diagnosis and management of fingertip injuries are vital to maintaining proper function of the hand and preventing permanent disability. A subungual hematoma is a painful condition that involves bleeding beneath the nail, usually after trauma. Treatment requires subungual decompression, which is achieved by creating small holes in the nail. A nail bed laceration is treated by removing the nail and suturing the injured nail bed. Closed fractures of the distal phalanx may require reduction but usually are minimally displaced and stable, and can be splinted. Open or intra-articular fractures of the distal phalanx may warrant referral. Patients with mallet finger cannot extend the distal interphalangeal joint because of a disruption of the extensor mechanism. Radiographs help to differentiate between tendinous and bony mallet types. Most mallet finger injuries heal with six to eight weeks of splinting, but some require referral. Flexor digitorum profundus avulsion always requires referral. Dislocations of the distal interphalangeal joint are rare and usually occur dorsally. PMID- 11388711 TI - When to suspect and how to monitor babesiosis. AB - In the past decade, cases of babesiosis in humans have been reported with increasing frequency, especially in the northeastern United States. Babesia microti (in the United States) and bovine strains (in Europe) cause most infections in humans. Most cases are tick-borne, although cases of transfusion associated and transplacental/perinatal transmission have also been reported. Factors associated with more severe disease include advanced age, previous splenectomy and immunodeficient states. Symptoms include high fever, chills, diaphoresis, weakness, anorexia and headache. Later in the course of the illness, the patient may develop jaundice. Congestive heart failure, renal failure and acute respiratory distress syndrome are the most common complications. Therapy using the combination of quinine sulfate and clindamycin was the most commonly used treatment; however, atovaquone suspension plus azithromycin was recently reported an equally effective and less toxic therapy. Exchange transfusion, together with antibabesial chemotherapy, may be necessary in critically ill patients. PMID- 11388712 TI - Information from your family doctor. What is babesiosis? PMID- 11388713 TI - New concepts in acute pain therapy: preemptive analgesia. AB - Pain, which is often inadequately treated, accompanies the more than 23 million surgical procedures performed each year and may persist long after tissue heals. Preemptive analgesia, an evolving clinical concept, involves the introduction of an analgesic regimen before the onset of noxious stimuli, with the goal of preventing sensitization of the nervous system to subsequent stimuli that could amplify pain. Surgery offers the most promising setting for preemptive analgesia because the timing of noxious stimuli is known. When adequate drug doses are administered to appropriately selected patients before surgery, intravenous opiates, local anesthetic infiltration, nerve block, subarachnoid block and epidural block offer benefits that can be observed as long as one year after surgery. The most effective preemptive analgesic regimens are those that are capable of limiting sensitization of the nervous system throughout the entire perioperative period. PMID- 11388714 TI - Information from your family doctor. Pain relief after surgery. PMID- 11388715 TI - Pneumococcal conjugate vaccine for young children. AB - Streptococcus pneumoniae causes approximately 3,300 cases of meningitis, 100,000 to 135,000 cases of pneumonia requiring hospitalization and 6 million cases of otitis media annually in the United States. Pneumococcal conjugate vaccine, approved in 2000 for use in the United States, was designed to cover the seven serotypes that account for about 80 percent of invasive infections in children younger than six years. This vaccine demonstrated 100 percent efficacy against invasive pneumococcal disease in the primary analysis of a large randomized, double-blind, controlled trial. In the follow-up analysis, performed eight months after the trial ended, efficacy against invasive disease was found to be 94 percent for the included serotypes. When initiated during infancy, the four-dose vaccination schedule is set at two, four, six and 12 to 15 months of age. The American Academy of Family Physicians recommends routine vaccination of infants, catch-up vaccination of children younger than 24 months and catch-up vaccination of children 24 to 59 months of age with high-risk medical conditions such as sickle cell disease and congenital heart disease. PMID- 11388716 TI - Information from your family doctor. Pneumococcal conjugate vaccine: what a parent needs to know. PMID- 11388717 TI - Bicycle-related injuries. AB - Bicycle riding is a popular form of recreation among persons of all ages, and related injuries cause significant morbidity and mortality. Most injuries occur in males and are associated with riding at high speed; most serious injuries and fatalities result from collisions with motor vehicles. Although superficial soft tissue injuries and musculoskeletal trauma are the most common injuries, head injuries are responsible for most fatalities and long-term disabilities. Overuse injuries may contribute to a variety of musculoskeletal complaints, compression neuropathies, perineal and genital complaints. Physicians treating such patients should consider medical factors, as well as suggest adjusting various components of the bicycle, such as the seat height and handlebars. Encouraging bicycle riders to wear helmets is key to preventing injuries; protective clothing and equipment, and general safety advice also may offer some protection. PMID- 11388718 TI - Information from your family doctor. Tips on bike safety. PMID- 11388719 TI - Significant FDA approvals in 2000. PMID- 11388720 TI - IDSA issues guidelines on the treatment of Lyme disease. Infectious Diseases Society of America. PMID- 11388721 TI - Ambient dose equivalent rate in Goiania 12 years after the 137Cs radiological accident. AB - This paper describes the situation of ambient dose equivalent rates in four of the main foci of 137Cs contamination in the city of Goiania, Brazil, in 1999, 12 y after one of the worst radiological accidents in the world. During the decontamination, all the buildings of the three highly contaminated sites were demolished and the top soil removed. Afterwards, the soil of two of these lots was covered with concrete, and they remain vacant today. The soil of the third of these lots, identified here as E, previously known as junkyard II, was covered only with clean soil. Three to four years after the accident, new houses were constructed on this lot, and some very poor people live and work there collecting recyclable material. Gamma ray spectrometry, with a portable survey meter, was performed in the quoted places along with outdoor measurements in many other locations of Goiania. The average ambient dose equivalent rate due to natural background radiation from radionuclides in the soil and cosmic radiation in non contaminated areas of the city of Goiania is 62 nSv h(-1). In most of the highly contaminated sites during the accident, the average ambient dose equivalent rate ranged from around 100 to 1,000 nSv h(-1). The only exception was site E, where values of ambient dose equivalent rate as high as 2.6 microSv h(-1) were found. PMID- 11388722 TI - Analysis of potential radiobiological effects related to a unified skin dose limit. AB - A unified skin dose limit of 0.5 Sv at a depth of 70 microm averaged over the highest 10 cm2 of skin exposed was evaluated to replace the existing limit of 0.5 Sv averaged over 1 cm2. This limit would apply to all exposures including non uniform exposures such as from hot particles on or off skin, skin contamination, or beams of charged particles or photons. The probabilities and severity of both stochastic and deterministic risks were estimated for a wide range of worst-case exposure scenarios using published radiobiological data and calculations of radial- and depth-dose distributions. Results indicate that exposures at the unified dose limit have the potential to cause effective doses of about 17 microSv (1.7 mrem), estimated stochastic risks of <3.3 x 10(-7) fatal skin cancers, and <1.6 x 10(-4) non-fatal skin cancers. The worst deterministic effects were estimated to be (a) based on a 2 Gy threshold, transient erythema induction to an area of 2.5 cm2 for uniform skin contamination over this same area and 0.65 cm2 for a 60Co hot particle 3 mm off of skin, (b) based on data for pig skin, 50% probability that 0.5 cm2 of skin would suffer 20% dermal thinning for uniform contamination with 106Rh spread over the same area, and (c) 10% probability of barely detectable transient acute necrosis or ulceration for 60Co or activated fuel particles 0.4 mm off of skin. It was concluded that the unified limit would provide a more logical system of dose control with possible savings of whole-body dose and other benefits. PMID- 11388723 TI - Former radiation worker Medical Surveillance Program at Rocky Flats. AB - The United States Department of Energy (DOE), Office of Occupational Medicine and Medical Surveillance, has supported an ongoing Former Radiation Worker Medical Surveillance Program at the DOE Rocky Flats site since 1992. The program currently is managed for DOE by Oak Ridge Institute for Science and Education through a contract with Oak Ridge Associated Universities. Participation in the program is entirely voluntary and provides former Rocky Flats workers who were exposed to radiation with long-term medical monitoring and an update to the assessment of their radiation dose. Program participants receive medical examinations and in vivo and in vitro bioassay measurements of residual radioactivity. Radiation doses to participants are largely a result of internal depositions of plutonium and its radioactive decay products. The causes of many of the higher internal doses were accidents that generally are well documented. Former radiation workers are invited to participate in the program if they meet specific criteria for radiation exposure. Informed consent is documented using a consent form approved by an Institutional Review Board. Demographic, medical, and dosimetric information is maintained in a computer database and will be evaluated for any trends or correlations between exposure and health outcome. PMID- 11388724 TI - Lifetime risk of lung cancer due to radon exposure projected to Japanese and Swedish populations. AB - Lifetime risk projections depend greatly on both background lung cancer rates and the selection of the risk model. Since background lung cancer rates differ from subject populations and the time, etiological risk of lifetime lung cancer mortality per unit radon exposure in WLM should be estimated for each subject population and the time of interest. To answer quantitatively how much are the differences among the projected risks for different populations, the Swedish case control-study-based risk projection model was applied to the Japanese and Swedish populations from 1962 to 1997 as subject populations because of their distinct trends of lung cancer rates. To compare the results with the reference population and authorized risk projection models, U.S. population 1997 and the two risk projection models in BEIR VI report were applied, respectively. Lifetime risk of lung cancer mortality projected for Japanese, Swedish, and U.S. populations in 1997 per radon progeny exposure were estimated to range from 1.50 (0.40-3.19) x 10(-4) WLM(-1) to 9.86 (2.62-20.9) x 10(-4) WLM(-1), which could be compared to the detriment associated with a unit effective dose. Conclusive dose conversion coefficients in this study ranged from 2.05 (0.55-4.37) to 13.5 (3.59-28.6) mSv WLM(-1), and within this range the discrepancy between dosimetric and epidemiological approaches was included. PMID- 11388725 TI - Statistical data evaluation in mobile gamma spectrometry: an optimization of on line search strategies in the scenario of lost point sources. AB - There is a potential risk that hazardous radioactive sources could enter the environment, e.g., via satellite debris, smuggled radioactive goods, or lost metal scrap. From a radiation protection point of view there is a need for rapid and reliable methods for locating and identifying sources. The methods could also be used to locate hot spots after radioactive fallout. Carborne and airborne gamma spectrometry systems are suitable for the task. This work focuses on a situation where the radionuclide to search for is known, which is not an unlikely scenario. The possibility that the source is located near a road can be high, and thus motivating a carborne spectrometer system. The main object is to optimize on line statistical methods in order to achieve a high probability for locating the point source and still have reasonably few false alarms caused by variations in the natural background radiation. Data were obtained from a carborne 3-L NaI(Tl) detector and two point sources located at various distances from the road. The nuclides used were 137Cs and 131I. Spectra were measured stationary on the road. From these measurements we have reconstructed counts in spectral windows applicable to different speed and sampling times; the time 3 s and speeds 32 and 54 km h(-1) are used in this work. The maximum distance a source can be located from the road and still be detected is estimated with four different statistical analysis methods. This distance is called the critical distance, CD. The method is applied on gross counts in the full energy peak spectral window. For each method alarm levels have been calculated from background data obtained in Scania (Skane), in the south of Sweden. The results show large differences in CD. With the best approach, the two sources could be detected from about 180 m (137Cs, 6 GBq) and 170 m (131I, 4.5 GBq). PMID- 11388726 TI - Dose absorbed by mouse red bone marrow after oral administration of tritiated water. AB - Tritiated water (HTO) was given to mice orally. Dose accumulation patterns in various organs and tissues after oral HTO intake were compared with those after intraperitoneal injection. The accumulated dose was 10-20% higher after intraperitoneal injection than after oral administration. A new technique was developed to isolate mouse red bone marrow from tibia. The absorbed beta-ray dose in the red bone marrow after oral tritium intake was lower than the body-averaged dose that was estimated based on the tritium concentration in the urine or the blood and higher than the absorbed dose in the liver and testis. PMID- 11388727 TI - Conservative evaluation of space radiation dose equivalent using the glow curve of 7LiF:Mg, Ti (TLD-700). AB - For conservative evaluation of dose equivalent in space, a simple method using two glow peaks in TLD-700 has been proposed. This method is superior to the method using the two-peak ratio in terms of following the 1990 ICRP recommendation. Dependence of each peak on LET was confirmed using relativistic heavy-ion beams (He, C, Ne, Ar, and Kr) at NIRS-HIMAC. TL values as gamma-ray absorbed-dose equivalent of both peak areas were additionally combined to conservatively estimate a dose equivalent over a range of LET of 0.5-440 keV microm(-1). This method was tested in an 8.8-d Shuttle-Mir mission (STS-89) at 400 km x 51.65 degrees. The dose-equivalent rates obtained at two positions in the Spacehab module were about 0.9 mSv d(-1); this result is reasonable in a conservative sense. PMID- 11388728 TI - Development and calculation of an energy dependent normal brain tissue neutron RBE for evaluating neutron fields for BNCT. AB - In Boron Neutron Capture Therapy (BNCT) of malignant brain tumors, the energy dependence of a clinically relevant Relative Biological Effectiveness (RBE) for epithermal neutrons, RBE(En), is important in neutron field design. In the first half of this paper, we present the development of an expression for the energy dependent normal-tissue RBE, RBE(En). We then calculate a reasonable estimate for RBE(En) for adult brain tissue. In the second half of the paper, two separate RBE expressions are developed, one for the RBE of the neutrons that interact in tissue via the 14N(n,p)14C reaction, denoted RBE(N), and one for the RBE of the neutrons which interact in tissue via the 1H(n,n')1H reaction, denoted RBE(H). The absorbed-dose-averaged values of these expressions are calculated for the neutron flux spectrum in phantom for the Brookhaven Medical Research Reactor (BMRR) epithermal neutron beam. The calculated values, [RBE(norm)N] = 3.4 and [RBE(norm)H] = 3.2, are within 6% of being equal, and support the use of equal values for RBEN and RBE(H) by researchers at Brookhaven National Laboratory (BNL). Finally, values of [RBE(norm)N] and [RBE(norm)H], along with the absorbed dose-averaged RBE for brain, [RBE(norm)b], were calculated as a function of depth along the centerline of an ellipsoidal head phantom using flux spectra calculated for our Accelerator-Based Neutron Source (ABNS). These values remained essentially constant with depth, supporting the use of constant values for RBE, as is done at BNL. PMID- 11388729 TI - The use of silicon photodiodes for radon and progeny measurements. AB - Reduced cost, high quantum efficiency and very good detection characteristics of PIN silicon photodiodes made possible their utilization for alpha particles detection. This paper presents different studies and applications of this type of detector for qualification and quantification of radon and its progeny in laboratory and in the field. Since photodiodes are sensitive to environmental electromagnetic fields, protecting cells were adapted for use around the detector. A small three-channel (2-5.5 MeV; 5.5-6.5 MeV; 6.5-8.2 MeV) portable alpha counter was developed that allows the determination of the radon concentration and the radon exhalation rate. Numerous field applications were conducted in different conditions (uranium waste storage, dwellings, etc.) where this simple and inexpensive instrumentation demonstrated very good performance, reliability, and easiness of operation while measuring radon concentrations from 50-100 Bq m(-3) to several MBq m(-3). PMID- 11388730 TI - Simulation of the radon and radon progeny detection by silicon photodiodes. AB - The use of silicon photodiodes for field radon measurements requires the utilization of measuring cells for protection against environmental electromagnetic fields. It is very important to study and optimize the impact of geometry conditions (dimensions of the measuring cell) on the photodiode detection efficiency that can be reduced more than 10-30 times. Two models (for volume-distributed nuclides around the detector and for radon progeny deposited on the diode surface radon progeny) were applied for simulation of the photodiode detection in measuring cells of different sizes. Their use allows an optimal choice of the most appropriate cell regarding the measurement conditions. Thus, for soil gas measurements cells we use a cell of 3.6 cm in diameter by 20 cm in height; for determination of the radon exhalation rate we use a cell of 16 cm in diameter by 14 cm in height; and for measurements in dwellings and large spaces we use cells larger than 20 cm and lower than 4 cm. PMID- 11388731 TI - Unusual 232Th and 238U contamination on some road surfaces in Taoyuan, Taiwan. AB - At least eight bustling streets or approximately 3-5% of all the road surface of civilian utility in the downtown area of Taoyuan City, Taiwan, were inadvertently found to contain unusual levels of radioactivity during a routine environmental radiation surveillance in mid-1994. Crushed rock debris and coarse sands separated from the asphalt pavement were identified to be the source of excessive radioactivity. By employing gamma spectrometry, we have measured 232Th activity (via 228Ac) and mU activity (via 214Bi) in some of the samples to be up to about 4,000 and 1,000 Bq kg(-1), respectively. The dose rate on the road surface reached about 1.3 microSv h(-1), as compared with the background level of 0.08 microSv h(-1) in much of Taiwan. This unusual radioactivity was due to accidental mixing of road construction materials with materials enriched 232Th and 238U. PMID- 11388732 TI - Estimation of dietary 232Th, 238U, cesium, and strontium intakes in Vietnamese people from different geographical regions. AB - Diet samples were collected to estimate dietary intakes of 232Th, 238U, cesium, and strontium for Vietnamese adults using duplicate portion studies and market basket studies. Average concentrations of 232Th, 238U, cesium, and strontium in diet samples were 4.0 ng/g-dry, 2.5 ng/g-dry, 41 ng/g-dry, and 4.5 ng/g-dry, respectively. Daily intakes per person for 232Th, 238U, cesium, and strontium were estimated to be 0.99 microg, 0.66 microg, 10 microg, and 1.2 mg, respectively. Daily intakes of 232Th, 238U, and cesium differed statistically between northern and southern of Vietnam, depending on geological conditions and food habits. However, intakes of the four nuclides were similar to the global averages. PMID- 11388733 TI - Contribution from thoron on the response of passive radon detectors. AB - In order to evaluate the reliability of measured values of radon concentration, a thoron sensitivity test for passive radon detectors was carried out. To do this test, a thoron chamber system was first set up. The system consists of four parts: an exposure chamber, a gas generator, an environmental monitor, and a measuring device. Five types of radon detectors were examined using the chamber system. After connecting the exposure chamber with the gas generator through an external pump, thoron gas was circulated through the system. The detectors were exposed to thoron-rich air for several days. The mean ratio between thoron and radon concentrations throughout the exposure period was 10:1. Some of the detectors provided values different from the actual radon concentration. Although the presence of thoron can be negligible in most cases, it is necessary to check the thoron contribution to the detector response with the proposed or similar test before practical use. PMID- 11388734 TI - A simple calculation for the buildup and decay of radon progeny. AB - A Markov chain in conjunction with an electronic spreadsheet is used to generate the buildup and decay patterns of radon progeny in a procedure involving only multiplication and addition. Markov chains are highly suitable for calculations of this type. Because the Markov matrix may not be readily available, and because some health physicists may not be familiar with this procedure, we give the values of a Markov matrix and demonstrate how it can be used to produce a Markov chain describing the buildup and decay of radon progeny. PMID- 11388736 TI - Bayesian formulations are not appropriate for a priori specification of analytic detection capabilities. PMID- 11388735 TI - Plutonium dose assessments based on a new model derived from ICRP 67. PMID- 11388737 TI - Structural and functional features of the CD34 antigen: an update. AB - CD34 is a heavily glycosylated type I transmembrane molecule, that can be phoshorylated by a variety of kinases including Protein kinase C and Tyrosine kinases. The classification of epitopes detected by different CD34 MAbs has aided the selection of appropriate antibodies for use in specific clinical and research laboratory settings. Detailed structural analyses and cloning studies have confirmed that CD34 is a sialomucin, and have suggested that the fine composition of the carbohydrate moieties contained in its extended N-terminal region is important in determining its interactions with a variety of different ligands. For high endothelial venules (HEV) CD34 to serve as a ligand for L-selectin, the O-linked glycans of HEV CD34 are modified in an exquisitely specific manner with a variety of sialyl- and sulfo-transferases. In contrast, CD34 is not the ligand for L-selectin in hematopoietic stem/progenitor cells (HSPCs) and despite much endeavour, ligands for hematopoietic CD34 remain to be identified. PMID- 11388738 TI - CD133. PMID- 11388739 TI - CD135. PMID- 11388740 TI - Flow cytometric enumeration and immunophenotyping of hematopoietic stem and progenitor cells. AB - Flow cytometric enumeration of CD34+ hematopoietic stem and progenitor cells (HPC) is widely used for evaluation of graft adequacy of peripheral blood stem cell grafts, and is also useful in planning the apheresis sessions necessary to obtain these grafts. The state-of-the-art method to enumerate CD34+ cells makes use of a multiparameter definition of HPC based on their light scatter characteristics and dim expression of CD45, and the use of counting beads to derive the concentration of CD34+ cells directly from the flow cytometric assessment. This method can be extended with a viability stain and additional markers for further immunological characterization of CD34+ cells, and has been successfully implemented in multicenter trials. Thus, the lower threshold of a safe HPC graft in terms of short- and long-term hematopoiesis may be more accurately defined. PMID- 11388741 TI - Assays for the assessment of human hematopoietic stem cells. PMID- 11388742 TI - Bone marrow mesenchymal cells: biological properties and clinical applications. PMID- 11388743 TI - Different growth factor requirements for the ex vivo amplification of transplantable human cord blood cells in a NOD/SCID mouse model. AB - The growth factor combination containing early acting cytokines FLT-3 ligand (FL), Stem Cell Factor (SCF) and thrombopoietin (TPO) is able to maintain, for an extended culture period, early stem cells, defined as long-term repopulating NOD/SCID mice (Scid Repopulating Cell-SRC) contained in cord blood (CB). In this culture system, the role of IL-6 and IL-3 has not been clearly established. Using a combination of FL+TPO+SCF with or without IL-6, we were able to form CB CD34+ cells for 30 weeks. The CB CD34+ cells cultured in this system engrafted NOD/SCID mice after 6 weeks of culture; the cells from primary recipients were also able to engraft secondary NOD/SCID mice. When CB CD34+ cells were cultured in the presence of IL-3 in the place of IL-6 we observed an even better expansion of cells and a similar clonogenic progenitor output in the first 8 weeks of culture. However, more primitive LTC-IC output increased up to week 6 with the growth factor combination containing IL-3 and then decreased and disappeared, while with the growth factor combination with or without IL-6 increased up to week 23. Cells cultured for 4 weeks with the 4-factor combination containing IL-3 engrafted NOD/SCID mice less efficiently. Repopulation of NOD/SCID mice was no longer observed when ex vivo expansion was performed for 6 weeks. This study provides some evidence that no differences could be detected in long-term maintenance and even expansion of human primitive cord blood cells cultured with FL+TPO+SCF in the presence or absence of IL-6. Under the culture conditions employed in this study, the presence of IL-3 reduced the repopulating potential of expanded CB CD34+ cells. PMID- 11388745 TI - Immunophenotypic characteristics of PB-mobilised CD34+ hematopoietic progenitor cells. PMID- 11388744 TI - Dendritic cell differentiation from hematopoietic CD34+ progenitor cells. AB - Dendritic cells (DC) are the most powerful antigen presenting cells (APC) and play a pivotal role in initiating the immune response. In light of their unique properties, DC have been proposed as a tool to enhance immunity against infectious agents and in anticancer vaccine strategies. In the last few years, the development of DC has been extensively investigated. The present paper summarizes the most recent findings on the differentiation of myeloid DC from hematopoietic CD34+ progenitors and methods for DC generation in vitro. A better understanding of DC function has important implications for their use in clinical settings. PMID- 11388746 TI - Importance of CD34+ cell subsets in autologous PBSC transplantation: the mulhouse experience using CD34+CD38- cells as predictive tool for hematopoietic engraftment. AB - Over the years, various biological parameters have been proposed for predicting rapidity and long term maintenance of hematopoietic engraftment after peripheral blood stem cell transplantation (PBSCT). Determination of the graft content in CFU-GM was the only one available until the end of the eighties. But, for technical reasons, and also because it does not actually evaluate the self renewal potential of the cell products reinfused, it has now been commonly replaced by the determination of CD34+ cell amounts, which are known to contain the pluripotent hematopoietic stem cells. However, a frequent discrepancy still exists between the number of CD34+ cells reinfused and the engraftment efficiency. We have recently demonstrated a higher accuracy of the numbers of CD34+38- cells contained in graft products to predict rapidity of trilineage engraftment, which has further been confirmed by other investigators. Furthermore, we and others, have proposed a threshold dose of 5 x 10(4) CD34+38- cells/kg b.w. below which the trilineage engraftment kinetics are significantly slower and unpredictible. This "cut-off" value also appears to be a realistic clinical tool to decide if hematopoietic growth factor(s) must be administered or not after PBSCT. Indeed, when for example, rh-G-CSF administration after transplant of CD34+38- amounts < 5 x 10(4) kg has indisputable positive effects on the rapidity of neutrophil engraftment, length of hospitalization and posttransplant costs, enough to make it fully justified in this situation, it is absolutely not the case when it is administered after reinfusion of CD34+38- cell amounts > 5 x 10(4) /kg. In this case, posttransplant rh-G-CSF administration could even result in a decrease in stem cells with self-renewal potential of the graft, which should still raise more concerns for its indiscriminate and costly use. PMID- 11388747 TI - Clinical and biological significance of CD34 expression in acute leukemia. PMID- 11388748 TI - Regulatory aspects and accreditation of the clinical use of hematopoietic progenitor cells in Europe. PMID- 11388749 TI - CD90. PMID- 11388750 TI - CD116 (granulocyte-macrophage colony stimulating factor receptor). PMID- 11388751 TI - CD117. PMID- 11388752 TI - CD122 (interleukin-2 receptor beta subunit). PMID- 11388753 TI - CD123 (interleukin 3 receptor alpha chain). PMID- 11388754 TI - Genetic discrimination in the workplace. PMID- 11388755 TI - Human genetics around the world. PMID- 11388756 TI - Improved detection of cystic fibrosis mutations in the heterogeneous U.S. population using an expanded, pan-ethnic mutation panel. AB - PURPOSE: To determine the comparative frequency of 93 CFTR mutations in U.S. individuals with a clinical diagnosis of cystic fibrosis (CF). METHODS: A total of 5,840 CF chromosomes from Caucasians, Ashkenazi Jews, Hispanics, African Americans, Native Americans, Asians, and individuals of mixed race were analyzed using a pooled ASO hybridization strategy. RESULTS: Sixty-four mutations provided a sensitivity of 70% to 95% in all ethnic groups except Asians, and at least 81% when the U.S. population was considered as a whole. CONCLUSIONS: For population based carrier screening for CF in the heterogeneous U.S. population, which is characterized by increasing admixture, a pan-ethnic mutation panel of 50 to 70 CFTR mutations may provide a practical test that maximizes sensitivity. PMID- 11388757 TI - Modifier locus for mitochondrial DNA disease: linkage and linkage disequilibrium mapping of a nuclear modifier gene for maternally inherited deafness. AB - PURPOSE: To examine the role of the nuclear genome in affecting the phenotypic expression of the simplest model of a mitochondrial DNA disease, maternally transmitted deafness. METHODS: Linkage analysis in families with maternally inherited deafness associated with the homoplasmic A1555G mutation. RESULTS: Significant linkage and linkage disequilibrium on chromosome 8 was identified. CONCLUSIONS: This finding represents the first identification of a modifier locus for a human mitochondrial DNA disease and supports the concept of mitochondrial DNA diseases having complex genetic inheritance. The eventual identification of this modifier gene will provide insights into the pathophysiological pathways determining the clinical expression of mitochondrial DNA diseases, an important step toward diagnostic and therapeutic interventions. PMID- 11388758 TI - Contribution of sickle cell disease to the occurrence of developmental disabilities: a population-based study. AB - PURPOSE: Population-based surveillance of children aged 3-10 years from metropolitan Atlanta was used to determine if stroke-related neurological damage in children with sickle cell disease (SCD) is associated with developmental disabilities (DD). METHODS: School and medical records were reviewed annually to identify eligible children. Observed-to-expected ratios, P values, and population attributable fractions were calculated. RESULTS: Children with SCD had increased risk for DD (O/E = 3.2, P < 0.0001), particularly mental retardation (O/E = 2.7, P = 0.0005) and cerebral palsy (O/E = 10.8, P < 0.0001). This risk was confined to DD associated with stroke (O/E = 130, P < 0.0001; for DD without stroke: O/E = 1.3, P = 0.23). CONCLUSIONS: Children with SCD have increased risk for DD associated with stroke; thus, aggressive interventions are needed to prevent stroke in these children. PMID- 11388759 TI - Life adaptation in 35 adults with sex chromosome abnormalities. AB - PURPOSE: This report from the last phase of the 36-year Denver Study provides information about the adult life adaptation of 35 men and women with sex chromosome abnormalities (SCA) followed since their identification at birth. METHODS: Sex chromatin screening of 40,000 consecutive newborns between 1964 and 1974 resulted in the cohort of 35 SCA men and women followed to date since birth. Sixteen chromosomally normal siblings served as controls. Data constituting this report was obtained from formal and informal interviews and psychological testing conducted in the final year of this study. RESULTS: The nonmosaic SCA propositi had lower mean intelligence quotients and psychosocial adjustment scores than did siblings. Consistent with these results, propositi also had lower levels of educational achievement and career success, although most have completed school, married, hold full-time employment, and are financially independent. CONCLUSIONS: While the SCA adults demonstrated lower levels of cognitive and psychosocial competence, their overall adaptation has been positive, particularly given an early literature suggesting a high rate of psychopathology and severe dysfunction. PMID- 11388760 TI - Clinical features of hereditary progressive arthro-ophthalmopathy (Stickler syndrome): a survey. AB - PURPOSE: To define variations in the clinical manifestations of Stickler syndrome. METHODS: A questionnaire was sent to 612 persons. RESULTS: Of the 316 usable replies, 95% of persons had eye problems (retinal detachment occurred in 60% of patients, myopia in 90%, and blindness in 4%); 84% had problems with facial structures such as a flat face, small mandible, or cleft palate; 70%, hearing loss; and 90%, joint problems, primarily early joint pain from degenerative joint disease. Treatment included cryotherapy and laser therapy for retinal detachment, repair of cleft palate, use of hearing and mobility aids, and joint replacements. CONCLUSIONS: There are wide variations of symptoms and signs among affected persons, even within the same family. There are delays in diagnosis, lack of understanding among family members, denial about the risk of serious eye problems, and joint disease. PMID- 11388761 TI - Genetic heterogeneity in autosomal dominant essential tremor. AB - PURPOSE: To perform linkage analysis of candidate loci in a large Midwestern family with autosomal dominant essential tremor. METHODS: Thirty-eight members of a six-generation family were evaluated for essential tremor using consensus criteria. Linkage analysis was performed with microsatellite markers reported for three genetic loci associated with familial essential tremor. RESULTS: Patients exhibited a combination of postural and kinetic tremor involving primarily the arms and hands, with a mean age of onset of 31 years. Genetic studies excluded linkage to ETM1 and ETM2 loci, as well as a candidate locus for parkinsonism and postural tremor on chromosome 4p. CONCLUSION: Familial essential tremor is a common hereditary movement disorder demonstrating phenotypic variability and genetic heterogeneity. PMID- 11388762 TI - Technical standards and guidelines for fragile X: the first of a series of disease-specific supplements to the Standards and Guidelines for Clinical Genetics Laboratories of the American College of Medical Genetics. Quality Assurance Subcommittee of the Laboratory Practice Committee. PMID- 11388764 TI - Establishing priorities in neurofibromatosis research: a workshop summary. AB - Progress in understanding the biology of the neurofibromatoses (NF1 and NF2) offers hope for the development of new, effective methods of treatment. In May 2000, the National Institute of Neurological Disorders and Stroke (NINDS) hosted a workshop that included leading researchers and clinicians from the NF community. The goal of the meeting was to assess current knowledge and identify priorities for future research. Needs identified included the development of better animal models, further study of the function of the NF1 and NF2 genes, and investigation of the role of modifier genes. The participants agreed that it will also be important to define further the natural history of NF1 and NF2 and to develop an infrastructure to support clinical trials. They also discussed the possible creation of research consortia and NF centers to promote the integration of basic and clinical research. PMID- 11388763 TI - American College of Medical Genetics statement of diagnostic testing for uniparental disomy. PMID- 11388766 TI - Portugal: the practice of medical genetics in Portugal. Portuguese Society of Human Genetics. PMID- 11388765 TI - Issues in human GenEthics. PMID- 11388767 TI - Greece: the Hellenic Association of Medical Geneticists. PMID- 11388768 TI - Smokeless tobacco and osteoporosis: a new relationship? AB - Osteoporosis is a common disorder of bone predominantly affecting postmenopausal women in which bone mineral density declines making bone more likely to break. Although cigarette smoking is a well-established risk factor for osteoporosis, smokeless tobacco (ST) use has never been suggested or evaluated as a risk factor for this bone disorder. This would be important to consider since in certain regions of the world, ST use is more prevalent than cigarette smoking, particularly among women. This paper reviews the animal and human evidence lending support to this new hypothesis, as well as the epidemiology of ST use that underscores the potential impact this modifiable behavior might have on osteoporosis world wide. PMID- 11388769 TI - Interferons: potential roles in affect. AB - A review of the literature on interferons was conducted and possible roles in neuropsychiatric disorders with affective disturbances are assessed. Interferons and interferon receptors are present in the limbic system where they appear to exert physiological effects pertinent to affect, most potently when levels rise during CNS infections. Interferons interact closely with cytokines and nitric oxide, signaling molecules implicated in depression. Results from knock-out mice suggest a role for interferon-gamma in moderating fear and anxiety, while other lines of evidence point to a role in arousal and circadian rhythms. The interferon-alpha receptor deploys an arginine methyltransferase affecting RNA editing and splicing, which seem to be disrupted in schizophrenia and bipolar disorder. S-Adenosylmethionine (SAMe), an effective antidepressant, may owe its effects in the latter disorders in part to variations in the strength of interferon-alpha signaling impacting RNA processing. Antiviral effects of interferons are of interest in lieu of viral theories of affective disorders. Finally, the relative levels of interferons gamma and alpha might play important roles in neural, and glial, development, as well as the dialog between the CNS and the immune system. PMID- 11388770 TI - Panax ginseng and Eleutherococcus senticosus may exaggerate an already existing biphasic response to stress via inhibition of enzymes which limit the binding of stress hormones to their receptors. AB - A mechanism of action for Panax ginseng (PG) and Eleutherococcus senticosus (ES) is proposed which explains how they could produce the paradoxical effect of sometimes increasing and sometimes decreasing the stress response. The mechanism suggests that this biphasic effect results from increased occupancy of positive and negative feedback stress hormone receptors by their natural ligands due to inhibition of specific enzymes which function to limit receptor occupancy. Specifically, it is suggested that PG inhibits 11-beta hydroxysteroid dehydrogenase one and ES inhibits catechol- O -methyl transferase, both of which reside in close proximity to stress hormone receptors and catalyse the degradation of stress hormones into inactive compounds. In addition, it is suggested that the increased energy said to result from PG and ES may be a consequence of their increasing the occupancy of stress hormone receptors which function to redistribute the body's energy reserves from regeneration to activity. PMID- 11388771 TI - E-Cadherin, beta -Catenin and cadmium carcinogenesis. AB - Cadmium (Cd(2+)) is an important industrial and environmental pollutant that has been classified as a human carcinogen. Studies reported in the literature indicate that cadmium may play a role in both the initiation of cancer, by activating oncogenes, and in the progression of cancer, by increasing the metastatic potential of existing cancer cells. However, the mechanisms underlying these effects have yet to be elucidated. Recent studies from our laboratory have shown that cadmium can disrupt the tight junctions between many types of epithelial cells by interfering with the normal function of E-cadherin, a Ca(2+) dependent cell adhesion molecule that plays a key role in epithelial cell-cell adhesion. This finding may be especially significant because a large volume of evidence indicates that the disruption of E-cadherin-mediated cell adhesion can trigger the beta-catenin-mediated activation of oncogenes in epithelial cells and increase the invasive potential of existing epithelial-derived cancers. The hypothesis that we are proposing is that the cadmium-induced disruption of E cadherin dependent cell-cell junctions may represent a pivotal step in both the initiation of cancer by cadmium and in the tumor promoting actions of cadmium. PMID- 11388772 TI - Repression of cellular anaplerosis as the hypothesized mechanism of gamma linolenic acid-induced toxicity to tumor cells. AB - In in vitro cultures, the cell is virtually isolated and can no longer rely on mechanisms for physiological regulation of substrate availability found in tissues. More emphasis is laid on utilization of preponderant substrate in a proposed reciprocal relationship between glycolysis and free fatty acid (FFA) oxidation for energy. Supraphysiological concentrations of gamma-linolenic acid and some other polyunsaturated fatty acids (PUFAs) therefore suppress glycolysis but also inhibit FFA oxidation initiated through a cytochrome P450-mediated epoxidation of PUFA to inhibit fatty acid synthase (FAS) activity. FAS inhibition accumulates malonyl CoA which inhibits carnitine palmitoyl transferase I and prevents FFA oxidation. The cell is starved of energy and anabolic intermediates, leading to decreased proliferation or death for tumor cells. Tumor cells are more vulnerable to this induced toxicity due to possession of specific phenotypes of elevated expression for FAS and pyruvate kinase, type M2, both factors inducing tumor cell apoptosis on inhibition. PMID- 11388773 TI - Consumption of carrageenan and other water-soluble polymers used as food additives and incidence of mammary carcinoma. AB - This study examined the hypothesis that the increasing incidence of mammary carcinoma in the USA in the twentieth century may be related to the consumption of carrageenan and possibly other water-soluble polymers. Widely used as food additives in the Western diet, the water-soluble polymers, also known as gums, are generally regarded as inert. However, the gum carrageenan which is comprised of linked, sulfated galactose residues has potent biological activity and undergoes acid hydrolysis to poligeenan, an acknowledged carcinogen. A time-trend analysis using age-adjusted incidence data and consumption data from established sources tested the hypothesis that increased consumption of the gums may be associated with increased incidence of mammary carcinoma. Correlations were determined using Pearson and Spearman correlation coefficients, incorporating lag intervals of 10 to 35 years. This analysis demonstrated that increasing consumption of several gums correlates positively with increased incidence of breast carcinoma. PMID- 11388774 TI - A model of hydraulic interactions in liver parenchyma as forces behind the intrahepatic bile flow. AB - The small diameters of bile canaliculi and interlobular bile ducts make it hard to attribute the bile flow solely to the process of secretion. In the model liver within its capsule is considered a limited space in which volume expansions of one part are possible only through the shrinking of other parts. The liver capsule allows only very slow volume changes. The rate of blood flow through the sinusoides is governed by the Poisseuill-Hagen law. The model is based on a concept of circulatory liver units. A unit would contain a group of acini sharing the same conditions of arterial flow. We can imagine them as an acinar group behind the last pressure reducer on one arterial branch. Acini from neighboring units compose liver lobules and drain through the same central venule. One lobule can contain acini from several neighboring circulatory units. The perfusion cycle in one unit begins with a transient tide in the arterial flow, governed by local mediators. Corresponding acini expand, grabbing the space by compressing their neighbors in the same lobules. Vascular resistance is reduced in dilated and increased in compressed acini. Portal blood flows through the dilated acini, bypassing the compressed neighbors. The cycle ends when the portal tide slowly diminishes and acinar volume is back on the interphase value until the new perfusion cycle is started in another circulatory unit. Each cycle probably takes minutes to complete. Increased pressures both in dilated and in compressed acini force the bile to move from acinar canalicules. Both up and down changes in acinar volume might force the acinar biliary flow. In cases of arterial vasoconstriction, increased activity of vasoactive substances would keep most of the circulatory units in the interphase and increased liver resistance can be expected. Liver fibrosis makes all acini to be of fixed volume and result in increased resistance. Because of that, low pressure portal flow would be more compromised, as reported. In livers without arterial blood flow, although some slow changes in the portal flows can be expected, acinar volume changes should be reduced. In acute liver injury, enlarged hepatocytes would diminish sinusoidal diameter and increase acinar resistance. In liver tumors, areas of neovascularization with reduced resistance would divert the arterial flow from the normal tissue, while in the compressed perifocal areas, increased vascular resistance should diminish mainly the portal flow. PMID- 11388775 TI - Metabolic disharmony and mortality. AB - The concept of 'metabolic harmony' is introduced and conceptualized as the state in which indices of metabolic activity (i.e., serum glucose, cholesterol, systolic and diastolic blood pressure, body mass index) within an individual attain their expected values given the individual values on related variables. Its complement, 'metabolic disharmony' (i.e., the extent to which an individual's 'profile' of metabolic variables is jointly unusual in relation to their expected values) is operationalized via Mahalanbis' D(2 )statistic calculated on these indices of metabolic activity (plus age and sex). Analysis of a large (N = 5209) longitudinal (32 years) cohort study shows that, independent of the linear and quadratic effects of the aforementioned metabolic variables, the disharmony index (DI) significantly and strongly predicted hazard of death (chi(2)(1) = 20.05, P < 0.00005). That is, each 10 percentile increase in DI was associated with a 6.9% increase in the hazard rate. The association of DI to hazard rate was not materially altered when potential confounders (e.g., smoking status) were added to the model or when all subjects were included by imputing missing data. These results demonstrate that metabolic disharmony is associated with, and may cause, an increased hazard of death. PMID- 11388776 TI - A new technique to elevate night time growth hormone release and a potential growth hormone feedback control loop. AB - A new technique for controllable elevation of night time growth hormone (GH) release in adult humans involves a synergy between oral intake of the naturally occurring compounds acetyl-L-carnitine (500 mg) and L-ornithine (25-100 mg) taken at night time sleep after a 3 to 4 hour fast. The set point for normal hypothalamic GH release appears to include a 'whole body' mitochondrial State 3 status 'feed back loop' controlled by systemic acetyl- L-carnitine levels. PMID- 11388777 TI - The loss of ego boundaries in schizophrenia: a neuromolecular hypothesis. AB - Virtually all genes of higher organisms have non-coding introns interspersed between the coding exons that must be spliced out in order to generate a mature messenger RNA molecule used to encode proteins. The hypothesis proposed in this paper is based on a new interpretation of molecular splicing as a primordial rejection mechanism that has evolved through time to serve as a molecular basis for other types of rejection, including rejection of thoughts and impulses in the human brain that are incompatible with environmental conditions. Psychiatrists generally explain delusions or hallucinations of schizophrenic patients in terms of 'loss of ego boundaries' or 'inner/outer confusion'. This new model proposes that the loss of the rejection function on the molecular level (non-splicing) in turn causes a loss of the boundary-setting function of glia in their interactions with the neural system. When introns are not spliced out from genes involved in neurotransmission, a variety of outcomes are possible. In some cases the production of neurotransmitters is decreased, in other cases the transmitter receptor has intron encoded sequence elements and/or premature stop codons that do not allow for proper transmitter occupancy. Because these 'chimeric' glial receptors cannot be occupied appropriately by their transmitters, the inactivating or boundary-setting function of synaptic transmission by glial cells is disturbed or lost. The inability to reject 'intronic ideas' may result in delusions and hallucinations, and could explain why schizophrenic patients are unable to test the reality of their ideas and are absolutely convinced that everything occurring in their brain is real. Finally, the concept of 'intronic ideas' from which schizophrenic patients are suffering is interpreted from an evolutionary point of view. It is proposed that many of the thoughts of schizophrenics have occurred too early in evolution and therefore cannot be realized, but in the future some of these thoughts will fit with reality as the environment changes and will no longer be considered delusions. PMID- 11388778 TI - Does the secretion and circulation of the cerebrospinal fluid really exist? AB - The secretion and circulation of cerebrospinal fluid have been studied in anaesthetized cats by means of a plastic cannula introduced into the aqueduct of Sylvius and by inspection of free escape of cerebrospinal fluid out of the end of the cannula. The fact that during the 120-minute period of observation not a single drop of CSF escaped out of the cannula, at physiological pressure, indicates that cerebrospinal fluid does neither secrete nor circulate. PMID- 11388779 TI - Non-haematological solid tumours as surrogate granulocytes: a possible mechanism for metastatic spread. AB - The concept that non-haematological solid tumours utilize normal leukocyte mechanisms, specifically those of granulocytes, to facilitate metastatic spread is presented here. In this article we will address the question of whether the process of metastasis is a phenomenon unique to tumour cells that utilize unique processes that are only found in neoplastic cell populations, or whether neoplastic populations 'turn on' or co-opt latent leukocyte genes normally not expressed by the parental, normal tissue of origin. We suggest that ectopic expression of leukocyte genes in epithelial tumours may be the simplest method for solid tumours to acquire the mechanisms for neoplastic spread through metastasis and may also account for the observed lack of host immunosurveillance of such tumours. PMID- 11388780 TI - Natriuresis of fasting: the possible role of leptin-neuropeptide Y system. AB - We developed a new hypothesis claiming that natriuresis of fasting is not only caused by diminished insulin resistance and hyperinsulinaemia with the subsequent reduction of renal sodium retention but it can also be attributed to the function of the leptin-NPY system. Each element of this concept has been substantiated by convincing experimental evidences as follows:1. Leptin, the adipocyte-derived peptide hormone conveys information to the central nervous system about the size of body energy stores and it reciprocally regulates the hypothalamic expression of NPY, the major mediator of its metabolic and neuroendocrine actions.2. NPY has been demonstrated to be intimately involved in the regulation of renal functions; under various experimental conditions it increased urine flow rate and urinary sodium excretion presumably through stimulating the synthesis and/or release of other natriuretic factors.3. Fasting-induced suppression of tissue expression of leptin mRNA and circulating plasma leptin levels is associated with simultaneous activation of NPY system.4. This sequence of events implies that NPY contributes to natriuresis that occurs in response to fasting. PMID- 11388781 TI - Schizophrenia and liver dysfunction. AB - It is suggested that a non-hepatocellular liver dysfunction, caused by the presence of a congenital or acquired portal-systemic shunt, constitutes a major predisposing factor in the pathogenesis of schizophrenia. In addition to the common occurrence of schizophrenic reactions observed in liver disease, this suggestion is supported by autoptic findings in addition to the fact that a considerable number of abnormal biochemical and biological phenomena are shared by patients suffering from schizophrenia and portal-systemic shunting. The frequency of abnormal portal-systemic shunts in schizophrenia is unknown. Recent advances in non-invasive Doppler-sonographic techniques should enable an elucidation of this question. PMID- 11388782 TI - Upregulation of duodenal calcium absorption by poly-unsaturated fatty acids: events at the basolateral membrane. AB - Poly-unsaturated fatty acids, especially of the n-3 series, have a beneficial effect in treatment of osteoporosis in the elderly. Duodenal calcium absorption is a particularly vulnerable aspect of the development of this disease. It has been shown that the process of calcium transport through the rat duodenal enterocyte takes place in essentially three steps: entry of calcium through channels in the brush border (apical membrane), transcellular transport through the cytoplasm by calbindin and extrusion at the basolateral membrane by Ca(2+) ATPase and a Ca(2+)-Na(+)exchanger which is driven by Na(+), K(+)-ATPase. This paper presents a hypothesis that poly-unsaturated fatty acids can modulate both Ca(2+)-ATPase and Na(+), K(+)-ATPase activity either by a direct action on the enzyme or by phosphorylation processes via protein kinases A and C and thus exert their positive influence on calcium absorption in this manner. PMID- 11388783 TI - Micronutrient accumulation and depletion in schizophrenia, epilepsy, autism and Parkinson's disease? AB - Zinc has several crucial functions in brain development and maintenance: it binds to p53, preventing it from binding to supercoiled DNA and ensuring that p53 cause the expression of several paramount genes, such as the one that encodes for the type I receptors to pituitary adenine cylase-activator peptide (PACAP), which directs embryonic development of the brain cortex, adrenal glands, etc.; it is required for the production of CuZnSOD and Zn-thionein, which are essential to prevent oxidative damage; it is required for many proteins, some of them with Zn fingers, many of them essential enzymes for growth and homeostasis. For example, the synthesis of serotonin involves Zn enzymes and since serotonin is necessary for melatonin synthesis, a Zn deficiency may result in low levels of both hormones. Unfortunately, Zn levels tend to be low when there is excess Cu and Cd. Moreover, high estrogen levels tend to cause increased absorption of Cu and Cd, and smoking and eating food contaminated with Cd result in high levels of the latter. Furthermore, ethanol ingestion increases the elimination of Zn and Mg (which acts as a cofactor for CuZnSOD). Increased Cu levels may also be found in people with Wilson's disease, which is a rather rare disease. However, the heterozygote form (only one faulty copy of the chromosome) is not so rare. Therefore, the developing fetus of a pregnant women who is low in Zn and high in Cu may experience major difficulties in the early development of the brain, which may later manifest themselves as schizophrenia, autism or epilepsy. Similarly, a person who gradually accumulates Cu, will tend to experience a gradual depletion of Zn, with a corresponding increase in oxidative damage, eventually leading to Parkinson's disease. Also discussed are the crucial roles of histidine, histamine, vitamin D, essential fatty acids, vitamin E, peroxynitrate, etc. in the possible oxidative damage involved in these mental diseases. PMID- 11388784 TI - How many deleterious mutations are there in the human genome? AB - An estimate of the number of deleterious mutations in the human genome is made using data on the frequency of rare recessive disease in cousin marriages and in the general population. Sexual reproduction ensures that deleterious mutations are distributed at random in zygotes with an approximate Poisson distribution. The mean of this distribution is the sum of the mean number of deleterious mutations in zygotes which contribute to the next generation (Y) and the mean number of new mutations which arise in each human generation (X). The estimates are that X is between 1 and 2.6 and Y is between 12 and 32. A mathematical model based on redundancy is then used to predict how zygote survival will vary with the number of deleterious mutations. The form of this relationship is the same as that seen in experiments on cell survival following radiation-induced mutational damage and this provides independent support for this theoretical approach. The zygotes that survive to contribute to the next generation have a skewed distribution with a mean of Y. It is argued that the number of deleterious mutations in the genome is an important variable in health and disease. PMID- 11388785 TI - Some semen abnormalities may cause infertility by impairing implantation rather than fertilization. AB - The hypo-osmotic swelling (HOS) test measures the functional integrity of the sperm membrane. Although, the sperm membrane is essential for the fertilization of oocytes, several clinical studies suggest that abnormally low HOS test scores do not predict poor or failed fertilization in human in-vitro fertilization trials. However, in-vivo and in-vitro studies clearly demonstrate that a low HOS score is associated with poor pregnancy rates suggesting this sperm defect causes implantation problems rather than fertilization problems. The problem of implantation could be caused by the supernumerary sperm attached to the zona pellucida. Supporting evidence for this theory was demonstrated by finding high pregnancy and implantation rates despite low HOS scores following intracytoplasmic sperm injection which avoids the presence of supernumerary sperm on the zona pellucida. These data thus support theories that some sperm abnormalities may reduce fertility potential by causing implantation disorders rather than problems with fertilization. PMID- 11388787 TI - Polio, hepatitis B and AIDS: an integrative theory on a possible vaccine induced pandemic. AB - The hypothesis that simian virus 40 (SV40) infected polio vaccines may be linked to the evolution of acquired immunodeficiency disorder (AIDS), and certain cancers, has been advanced. Most recently, investigators discussed the likelihood of gene-reshuffling following SV40 infection as a precursor to acquired immune dysfunction. Findings of recent SV40 infections in four children born after 1982 suggest infections were transmitted vertically along gene lines. Earlier observations proved activation of a retrovirus gene by a hepatitis B virus (HBV) protein. This paper proposes a new integrative theory on the origin of AIDS. It advances the possibility of genetic recombinations with oncogene activation by HBV involving simian viruses that likely infected polio vaccinated blood donors to the initial hepatitis B (HB) vaccine trials conducted on gay men in New York City and Ugandan Blacks in the early to mid-1970s. The socio-economic and even military ramifications associated with this politically challenging thesis are discussed. PMID- 11388786 TI - The metabolism of sulphur in relation to the biochemistry of cystine and cysteine: its fundamental importance in biology. A cyclic interchange between their mono- and di-sulphides is the unique reaction creating life and intelligence. PMID- 11388788 TI - Iron, infection and the evolutionary ecology of heart disease. AB - Levels of iron intake and stored iron have been implicated as risk factors for coronary heart disease. More recently, considerable interest has centered on the role of a variety of infectious pathogens, particulary bacterial pathogens, in the development of artherosclerosis and heart disease. The mechanism whereby elevated iron levels increase the risk of coronary heart disease is not well understood. Here it is proposed that the influence of iron levels on the persistence, pervasiveness and intensity of bacterial infections may play an important role in the development of coronary heart disease. PMID- 11388789 TI - Cyclosporine therapeutic and toxic effects may be related to different cyclosporine concentration zones in plasma. AB - Different cyclosporine concentration zones can exist in plasma due to the temperature dependency in distribution and association, therefore cyclosporine therapeutic and toxic effects may partially be related to these concentration zones. PMID- 11388790 TI - Repression of the enzyme lactate dehydrogenase and the spontaneous remission or regression of cancer. AB - Lactic acid produced by the malignant tumor is what keeps the malignancy going. Suppression or the removal of lactic acid is the prerequisite for any cancer treatment. PMID- 11388793 TI - Immunoaffinity purification and reconstitution of human alpha(2)-adrenergic receptor subtype C2 into phospholipid vesicles. AB - Large quantities of correctly folded, pure alpha(2)-adrenergic receptor protein are needed for structural analysis. We report here the first efficient method to purify human alpha(2)-adrenergic receptor subtype C2 to homogeneity from recombinant yeast Saccharomyces cerevisiae by one-step purification using a monoclonal antibody column (specific for alpha(2)C2). We show that the adrenoceptor antagonist phentolamine stabilized the receptor during purification. We used a very effective chaotropic agent, NaSCN, to elute the receptor from the immunoaffinity column with an overall yield of 34% before reconstitution. Ligand binding of detergent-solubilized, immunoaffinity-purified receptors could not be demonstrated, but partial recovery of ligand binding activity was achieved when purified receptors were reconstituted into phospholipid vesicles. The reconstituted receptors still bound radioligand after storage on ice for 4 weeks. This purification procedure can be easily scaled-up and thus demonstrates the utility of a monoclonal antibody column and NaSCN elution to purify large quantities of G-protein-coupled receptors. PMID- 11388794 TI - Expression and purification of monospecific and bispecific recombinant antibody fragments derived from antibodies that block the CD80/CD86-CD28 costimulatory pathway. AB - The development of recombinant techniques for rapid cloning, expression, and characterization of cDNAs encoding antibody (Ab) subunits has revolutionized the field of antibody engineering. By fusion to heterologous protein domains, chain shuffling, or inclusion of self-assembly motifs, novel molecules such as bispecific Abs can be generated that possess the subset of functional properties designed to fit the intended application. We describe the engineering of Ab fragments produced in bacteria for blocking the CD28-CD80/CD86 costimulatory interaction in order to induce tolerance against transplanted organs. We designed single-chain Fv antibodies, monospecific and bispecific diabodies, and a bispecific tetravalent antibody (BiTAb) molecule directed against the CD80 and/or CD86 costimulatory molecules. These recombinant Ab molecules were expressed in Escherichia coli, followed by purification and evaluation for specific interaction with their respective antigen in an enzyme-linked immunosorbent assay (ELISA). A specific sandwich ELISA confirmed the bispecificity of the bispecific diabodies and the BiTAb. PMID- 11388795 TI - Expression of Giardia duodenalis beta-tubulin as a soluble protein in Escherichia coli. AB - The beta-tubulin gene of the parasitic protozoan Giardia duodenalis has been expressed for the first time using a novel and direct method. The protein was expressed in both soluble and insoluble forms in an Escherichia coli-based expression system. The level of expression was found to be affected by several variables including the incubation temperature, length of time for which expression was carried out, and the E. coli culture volume. The protein expression system contributed no additional amino acids to the final fusion protein and the polyhistidine fusion sequence was easily removed from the beta tubulin protein using a specific enterokinase enzyme. The expression system also provided a means of preparing a soluble protein and purifying it by a relatively straightforward affinity chromatography method to give a very high level of protein purity. This makes the protein suitable for a number of applications for characterization including beta-tubulin antibody assays, alpha-/beta-tubulin binding regions, and beta-tubulin folding intermediates. PMID- 11388796 TI - Two-cistron system overexpression of chloroplast glyceraldehyde-3-phosphate dehydrogenase subunit B and B-derivatives from spinach in Escherichia coli. AB - A gene coding for the subunit B (GapB) of chloroplast glyceraldehyde-3-phosphate dehydrogenase from spinach and its two derivatives (GapBc) lacking the GapB specific C-terminal extension have been cloned by RT-PCR. These three genes have been overexpressed with full activity in Escherichia coli when a two-cistron expression system controlled by an inducible promoter P(trc) is used. With a suitable base composition of the first cistron, the expression level of GapB and the derivatives GapBc are expressed up to 15-20% of the total cell protein and around 20 mg of recombinant GapBcs with full activity are purified from 1 liter of cultured bacteria. The specific activity of the two derivatives GapBc (40-60 u/mg) is similar to that of GapA (50-70 u/mg) and lower than that of reported GapBc derivative (E. Baalmann, R. Scheibe, R. Cerff, and W. Martin, 1996, Plant Mol. Biol. 32, 505-513). PMID- 11388797 TI - One-step purification of a fully active hexahistidine-tagged human hexokinase type I overexpressed in Escherichia coli. AB - The conversion of glucose into glucose 6-phosphate (Glc 6-P)1 traps glucose in a chemical state in which it cannot leave the cell and hence commits glucose to metabolism. In human tissues there are at least three hexokinase isoenzymes responsible for hexose phosphorylation. These enzymes are constituted by a single polypeptide chain with a molecular weight of approximately 100 kDa. Among these isoenzymes, hexokinase type I is the most widely expressed in mammalian tissues and shows reversion of Glc 6-P inhibition by physiological levels of inorganic phosphate. In this work the hexokinase I from human brain was overexpressed in Escherichia coli, as a hexahistidine-tagged protein with the tag extending the C terminal end. An average of 900 U per liter of culture was obtained. The expressed protein was one-step purified by metal chelate affinity chromatography performed in NTA-agarose column charged with Ni(2+) ions. In order to stabilize the enzymatic activity 0.5 M ammonium sulfate was added to elution buffer. The specific activity of purified hexokinase I was 67.8 U/mg. The recombinant enzyme shows kinetic properties in agreement with those described for the native enzyme, and thus it can be used for biophysical and biochemical investigation. PMID- 11388798 TI - Systematic separation and purification of elastase, gelatinase (matrix metalloproteinase 9), and collagenase (matrix metalloproteinase 8) from polymorphonuclear leukocytes in dialyzers previously used by patients with renal failure. AB - We developed a simple and effective method for the systematic separation and purification of human polymorphonuclear leukocyte (PMN) proteinases, elastase, gelatinase (matrix metalloproteinase 9, type IV collagenase), and collagenase (matrix metalloproteinase 8), derived from the extracts of hollow fiber dialyzers that had been utilized in the treatment of patients with renal failure. The fraction containing elastase was grossly separated from that containing gelatinase and collagenase by heparin-Sepharose chromatography and purified in an aprotinin column. The remaining two enzymes were then separated using the gelatin Sepharose column after gel chromatography following ammonium sulfate precipitation. Gelatinase and collagenase were further purified by gelatin Sepharose chromatography as a latent form and by collagen-Sepharose chromatography as an activated form. This novel method offers procedural advantages over existing methods that separate PMNs from the whole blood of volunteers for experimental research purposes. PMID- 11388799 TI - Subcloning, expression, purification, and characterization of Haemophilus influenzae glycerol kinase. AB - Glycerol kinase (EC 2.7.1.30) is a bacterial sugar kinase and a member of the sugar kinase/actin/hsc-70 superfamily of enzymes. The enzyme from Escherichia coli is an allosteric regulatory enzyme whose activity is inhibited by fructose 1,6-bisphosphate (FBP) and the glucose-specific phosphocarrier of the phosphoenolpyruvate:glycose phosphotransferase system, IIA(Glc) (previously termed III(Glc)). Comparison of its primary structure with that of the highly similar Haemophilus influenzae glycerol kinase reveals that the amino acid sequence for the binding site for FBP is conserved while the amino acid sequence for the binding site for IIA(Glc) contains differences that are predicted to prevent its inhibition. To test this hypothesis, the H. influenzae glpK gene was assembled from DNA library fragments and subcloned into pUC18. The enzyme is expressed at high levels in E. coli. It was purified to greater than 90% homogeneity by taking advantage of its solubility behavior in a procedure that requires no column chromatography. The initial-velocity kinetic parameters of the purified enzyme are similar to those of the E. coli glycerol kinase. The H. influenzae glycerol kinase is inhibited by FBP but not by IIA(Glc), in agreement with the prediction based on sequence comparison. Sedimentation velocity experiments reveal that inhibition of HiGK by FBP is associated with oligomerization, behavior which is similar to EcGK. The possibility of utilizing mutagenesis studies to exploit the high degree of similarity of these two enzymes to elucidate the mechanism of allosteric regulation by IIA(Glc) is discussed. PMID- 11388800 TI - Recombinant expression of biologically active rat leptin in Escherichia coli. AB - Leptin is a 16-kDa nonglycosylated hormone that is produced in mature adipocytes and which acts primarily in the hypothalamus to reduce food intake and body weight. While the rat is a representative laboratory animal model in obesity research, so far recombinant rat leptin was not available. In the present study, rat leptin was recombinantly expressed in Escherichia coli and purified in a bioactive form to provide a further tool for the analysis of leptin functions in rats. Leptin cDNA was cloned by RT-PCR from total RNA of SD rat adipocytes, and overexpression was achieved by subcloning the leptin cDNA into the pET-29a vector, which enabled the recombinant expression of rat leptin as an S-peptide tagged fusion protein. Since the fusion proteins were expressed in inclusion bodies, after purification of the insoluble fraction, leptin proteins were refolded by sequential dialysis into physiological buffers. The biological activity of this recombinant protein was confirmed in proliferation assays using leptin-sensitive rat insulinoma cells as well as a newly developed leptin sensitive luciferase assay system. The specific binding of the S-tagged leptin to leptin-receptor-expressing cells was further shown by flow cytometry using fluorescence-conjugated S-proteins. PMID- 11388801 TI - The expression of a single-chain Fv antibody fragment in different plant hosts and tissues by using Potato virus X as a vector. AB - Some aspects of the expression of a single-chain Fv antibody fragment (scFv) driven by the plant viral vector Potato virus X (PVX) have been studied by quantitative ELISA. After inoculation of the infectious transcript, the vector was stable only for a few passages of sap transmission in the inoculated leaves of Nicotiana benthamiana and the reversal to wild type was more pronounced in the systemically invaded leaves. The amount of synthesized scFv varied when different solanaceous hosts were tested, being generally higher and less variable in inoculated than in systemically invaded leaves. In tomato and Datura stramonium the scFv was synthesized only in the inoculated leaves. The scFv was also synthesized in the PVX local hosts Chenopodium amaranticolor and C. quinoa. No correlation was found between PVX and scFv concentration in the inoculated and systemically invaded leaves of N. benthamiana and N. clevelandii. PMID- 11388802 TI - Expression and purification of Escherichia coli beta-glucuronidase. AB - A strong and constitutive expression vector of Escherichia coli beta glucuronidase with the isocitrate dehydrogenase promoter has been developed for producing a large amount of recombinant protein. More than 95% pure enzyme was obtained by a four step purification procedure-ammonium sulfate precipitation, DEAE ion-exchange chromatography, Superose 12 gel filtration, and hydroxyapatite steric ion-exchange chromatography. The overexpressed gene can produce 23 mg of pure enzyme from one liter of bacterial culture. PMID- 11388803 TI - Inexpensive purification of P450 reductase and other proteins using 2',5' adenosine diphosphate agarose affinity columns. AB - Two reductases, P450 oxidoreductase and P450Bm-3 reductase, were purified on a 2',5'-adenosine diphosphate solid support. Although the efficiency of these columns is well established, the cost of the resin and the eluting material 2' adenosine can be prohibitive. Herein we show that the less costly 2',3'-adenosine monophosphate is an excellent eluting material. PMID- 11388804 TI - Expression of mammalian geranylgeranyltransferase type-II in Escherichia coli and its application for in vitro prenylation of Rab proteins. AB - Mammalian geranylgeranyltransferase type II (GGTase-II) is a 100-kDa heterodimer that catalyzes the transfer of two 20-carbon geranylgeranyl groups from geranylgeranyl pyrophosphate onto C-terminal cysteine residues of Rab GTPases. This modification is essential for the biological activity of Rab proteins. Geranylgeranylation can be performed in vitro using recombinant GGTase-II but so far large-scale production of the enzyme was challenging. We report here the design of a two plasmid expression system that will produce GGTase-II at levels as high as 15 mg/L in Escherichia coli. The protein was produced as a heterodimer with the alpha subunit bearing a cleavable tandem 6His-glutathione S-transferase (GST) tag that was used for two-step purification of the enzyme. Purified enzyme was functionally active as determined by in vitro prenylation and phosphoisoprenoid binding assay. Furthermore, the GST-tagged GGTase-II was used for preparative in vitro prenylation of the Rab7:REP-1 complex. Using this procedure, 10 mg of doubly prenylated Rab7:REP-1 complex were obtained. PMID- 11388805 TI - Recombinant human tumor antigen MUC1 expressed in insect cells: structure and immunogenicity. AB - MUC1, a member of the mucin family of molecules, is a transmembrane glycoprotein abundantly expressed on human ductal epithelial cells and tumors originating from those cells. MUC1 expressed by malignant cells is aberrantly O-glycosylated. Differences in O-glycosylation of the tandem repeat region of MUC1 make tumor and normal forms of this antigen immunologically distinct. The tumor-specific glycoform is, therefore, expected to be a good target for immunotherapy and a good immunogen for generation of antitumor immune responses. We have generated a renewable source of this glycoform by expressing MUC1 cDNA in Sf-9 insect cells using a baculovirus vector. This form of MUC1 (BV-MUC1) is O-glycosylated at a very low level, approximately 0.3% (w/w), and this is not due to the lack of appropriate glycosylotransferases in insect cells. Peptidyl GalNAc-transferases isolated from Sf-9 cells were able to glycosylate in vitro a synthetic MUC1 peptide as efficiently as the transferases isolated from human milk. Neither preparation of peptidyl GalNAc-transferases, however, was able to glycosylate BV MUC1. This underglycosylated recombinant MUC1 mimics underglycosylated MUC1 on human tumor cells and could serve as an immunogen to stimulate responses that would recognize MUC1 on tumor cells. To test this we immunized mice with Sf-9 cells expressing BV-MUC1. Sera from immunized mice recognized MUC1 on human tumor cells. We also generated MUC1-specific T cells that proliferated in response to synthetic MUC1 peptide. PMID- 11388807 TI - Accurate disulfide formation in Escherichia coli: overexpression and characterization of the first domain (HF6478) of the multiple Kazal-type inhibitor LEKTI. AB - The human hemofiltrate peptide HF6478, a putative serine proteinase inhibitor, which is part of the precursor protein LEKTI, was cloned, overexpressed, and purified. HF6478 contains two disulfide bridges with 1-4, 2-3 connectivity, sharing partial homology to Kazal-type domains and other serine proteinase inhibitors. It was expressed as thioredoxin (Trx) fusion protein, and disulfide formation occurred in the oxidative cytoplasm of Escherichia coli Origami (DE3) strain which carries a trxB(-)/gor522(-) double mutation. The soluble fusion protein was purified using metal-chelating affinity chromatography. Cleavage of the Trx fusion protein with factor Xa and subsequent purification yielded the final product in amounts sufficient for structural studies. Characterization of recombinant HF6478 was done by amino acid sequencing, mass spectrometry, capillary zone electrophoresis, and CD spectroscopy. Taking the blood filtrate peptide HF6478 as example, we present a strategy which should facilitate the expression of different extracellular proteins in the E. coli cytoplasm. PMID- 11388806 TI - Refolding and purification of yeast carboxypeptidase Y expressed as inclusion bodies in Escherichia coli. AB - The genes encoding carboxypeptidase Y (CPY) and CPY propeptide (CPYPR) from Saccharomyces cerevisiae were cloned and expressed in Escherichia coli. Six consecutive histidine residues were fused to the C-terminus of the CPYPR for facilitated purification. High-level expression of CPY and CPYPR-His(6) was achieved but most of the expressed proteins were present in the form of inclusion bodies in the bacterial cytoplasm. The CPY and CPYPR-His(6) produced as inclusion bodies were separated from the cells and solubilized in 6 and 3 M guanidinium chloride, respectively. The denatured CPYPR-His(6) was refolded by dilution 1:30 into the renaturation buffer (50 mM Tris-HCl containing 0.5 M NaCl and 3 mM EDTA, pH 8.0), and the refolded CPYPR-His(6) was further purified to 90% purity by single-step immobilized metal ion affinity chromatography. The denatured CPY was refolded by dilution 1:60 into the renaturation buffer containing CPYPR-His(6) at various concentrations. Increasing the molar ratio of CPYPR-His(6) to CPY resulted in an increase in the CPY refolding yield, indicating that the CPYPR His(6) plays a chaperone-like role in in vitro folding of CPY. The refolded CPY was purified to 92% purity by single-step p-aminobenzylsuccinic acid affinity chromatography. When refolding was carried out in the presence of 10 molar eq CPYPR-His(6), the specific activity, N-(2-furanacryloyl)-l-phenylalanyl-l phenylalanine hydrolysis activity per milligram of protein, of purified recombinant CPY was found to be about 63% of that of native S. cerevisiae CPY. PMID- 11388808 TI - Construction and expression of an enzymatically active form of PECAM-1 containing the phosphatase domain of the protein tyrosine phosphatase, SHP-2. AB - Platelet endothelial cell adhesion molecule-1 (PECAM-1; CD31) is a 130-kDa transmembrane glycoprotein that is expressed on the surfaces of platelets, endothelial cells, and certain leukocyte subsets. The extracellular region of PECAM-1 contains six immunoglobulin homology domains, two of which (domains 1 and 2) mediate PECAM-1 homophilic interactions. Recent evidence suggests that a major function of the extracellular region of PECAM-1 is to determine its localization within the plane of the plasma membrane. The cytoplasmic domain of PECAM-1 contains an immunoreceptor tyrosine-based inhibitory motif that, upon tyrosine phosphorylation, supports recruitment of the Src homology 2 domain-containing protein tyrosine phosphatase, SHP-2. However, neither the targets of this PECAM 1/SHP-2 complex nor the significance of localizing SHP-2 to the borders of opposing PECAM-1-expressing cells is yet known. As a first step in addressing these issues, we designed a cDNA encoding a chimeric protein composed of the PECAM-1 extracellular domain fused to the phosphatase domain of SHP-2, which we call PECAM-1/PhD2. When immunopurified from stably transfected HEK293 cell lines expressing this recombinant protein, PECAM-1/PhD2 was found to possess constitutive enzymatic activity and appropriate border localization. This constitutively active chimeric protein will be useful in future studies designed to define the components of signal transduction pathways modulated by PECAM-1/SHP 2 signaling complexes. PMID- 11388809 TI - Overexpression, rapid isolation, and biochemical characterization of Escherichia coli single-stranded DNA-binding protein. AB - Escherichia coli (E. coli) single-stranded binding protein (SSB) is a valuable protein for various biotechnical applications, such as PCR and DNA sequencing. Here we describe an efficient expression and purification scheme where the tendency of SSB to aggregate at low salt concentration and high protein concentration is avoided. The method contains fewer steps of purification and results in high protein yield, compared to previous published protocols. In our protocol, cells are harvested after cultivation overnight and SSB is isolated by ammonium sulfate precipitation followed by anion-exchange chromatography. The yield from a 2-liter fed-batch fermentor is 2 g protein, which is higher than all production methods for SSB earlier reported. Moreover, the two classical isolation steps combined in the purification scheme are robust, cost-efficient, and suitable for scaling up. The resulting SSB is pure and a correctly folded tetramer with an apparent binding to single-stranded DNA with a K(D) of 10(-8) M, as determined by surface plasmon resonance. PMID- 11388810 TI - Systematic optimization of expression and refolding of the Plasmodium falciparum cysteine protease falcipain-2. AB - The Plasmodium falciparum cysteine protease falcipain-2 is a potential new target for antimalarial chemotherapy. In order to obtain large quantities of active falcipain-2 for biochemical and structural analysis, a systematic assessment of optimal parameters for the expression and refolding of the protease was carried out. High-yield expression was achieved using M15(pREP4) Escherichia coli transformed with the pQE-30 plasmid containing a truncated profalcipain-2 construct. Recombinant falcipain-2 was expressed as inclusion bodies, solubilized, and purified by nickel affinity chromatography. A systematic approach was then used to optimize refolding parameters. This approach utilized 100-fold dilutions of reduced and denatured falcipain-2 into 203 different buffers in a microtiter plate format. Refolding efficiency varied markedly. Optimal refolding was obtained in an alkaline buffer containing glycerol or sucrose and equal concentrations of reduced and oxidized glutathione. After optimization of the expression and refolding protocols and additional purification with anion-exchange chromatography, 12 mg of falcipain-2 was obtained from 5 liters of E. coli, and crystals of the protease were grown. The systematic approach described here allowed the rapid evaluation of a large number of expression and refolding conditions and provided milligram quantities of recombinant falcipain-2. PMID- 11388811 TI - Intein-mediated rapid purification of Cre recombinase. AB - Cre recombinase produced by bacteriophage P1 catalyzes site-specific recombination of DNA between loxP recognition sites in both prokaryotic and eukaryotic cells and has been widely used for genome engineering and in vitro cloning. Recombinant Cre has been overproduced in Escherichia coli and its purification involves multiple steps. In this report, we used an "intein" fusion system to express Cre as a C-terminal fusion to a modified protein splicing element, i.e., intein. The modified intein contained a Bacillus circulans chitin binding domain which allowed binding of the fusion protein on a chitin column and could be induced to undergo in vitro peptide bond cleavage which specifically released Cre from the column. Using the intein system, we have obtained highly pure nontagged Cre after just a single chromatographic step, which corresponded to approximately 80% recovery and 27-fold purification. The activity of the purified Cre was determined in an in vitro assay system and was found to remain stable over a period of more than 6 months. PMID- 11388812 TI - Expression in Spodoptera frugiperda (Sf21) insect cells of BT-R(1), a cadherin related receptor from Manduca sexta for Bacillus thuringiensis Cry1Ab toxin. AB - The cadherin-related receptor of Manduca sexta, BT-R(1), for the Cry1A family of Bacillus thuringiensis insecticidal toxins, was expressed in cultured Spodoptera frugiperda (Sf21) insect cells utilizing the expression vector deltaOp-gp64. Recombinant BT-R(1) was released by the Sf21 cells in soluble form into the culture medium and represents approximately 58% of total BT-R(1) produced by the cells. The soluble protein was purified by affinity chromatography using Cry1Ab toxin coupled to Sepharose 4B. The apparent molecular mass of purified soluble recombinant BT-R(1) is 195 kDa. Radiolabeled toxin bound to purified soluble BT R(1) with a K(d) value of 1.1 nM, which is similar to that of both membrane-bound BT-R(1) in Sf21 cells and natural BT-R(1) from M. sexta larval midgut tissue. Binding of radiolabeled toxin to soluble BT-R(1) was competitively inhibited by unlabeled Cry1Ab toxin but not by other Cry toxins as was observed also for membrane-bound BT-R(1). The recombinant soluble protein was stable in culture medium for at least 3 days at 27 degrees C and for 7 days at 4 degrees C and exhibited toxin-binding properties similar to the natural protein. Apparently, neither membrane association nor the extent of glycosylation influences the binding affinity and specificity of BT-R(1). Approximately 1 mg of purified BT R(1) was obtained per liter of insect cell culture supernatant, representing approximately 2 x 10(9) Sf21 cells. PMID- 11388813 TI - Isolation, expression, and characterization of fully functional nontoxic BiP/GRP78 mutants. AB - Mammalian BiP/GRP78 and Escherichia coli DnaK belong to the highly conserved hsp70 family and function as molecular chaperones in the endoplasmic reticulum or the cytosol, respectively. Induction of murine BiP/GRP78 expression in E. coli leads to growth arrest and cell death, independent of the bacterial strain and vector used. Analysis of various BiP constructs and mutants shows that the dominant-lethal phenotype is induced specifically by the expression of the 13.7 kDa C-terminal domain and abolished by a single substitution in that region. Deletion of that region also results in nontoxic gene products that can be overexpressed and purified to homogeneity. The nontoxic mutants are highly expressed in E. coli, representing up to 20% of the soluble fraction. They are catalytically active, depolymerize upon binding ATP or synthetic peptide, and interact with the J-domain of the DnaJ-like accessory protein, MTJ1, with near wild-type affinity. Our data indicate that the cytotoxic effect encountered during overexpression of recombinant proteins can be caused by a single domain and can be alleviated by a specific mutation or deletion in that region without altering the catalytic properties of the enzyme. PMID- 11388814 TI - Treatment with continuous positive airway pressure is not effective in patients with sleep apnea but no daytime sleepiness. a randomized, controlled trial. AB - BACKGROUND: The sleep apnea-hypopnea syndrome is defined by a pathologic number of respiratory events during sleep (the apnea-hypopnea index, defined as the number of apnea and hypopnea episodes per hour) and daytime symptoms (mostly, excessive sleepiness). In patients with the sleep apnea syndrome, treatment with continuous positive airway pressure (CPAP) normalizes both the apnea-hypopnea index and diurnal symptoms. However, the effect of CPAP in persons with a pathologic apnea-hypopnea index without daytime sleepiness is unclear. OBJECTIVE: To investigate the short-term effects of CPAP on quality of life, objective sleepiness, cognitive function, and arterial blood pressure in nonsleepy patients with a pathologic apnea-hypopnea index. DESIGN: Multicenter randomized, placebo controlled, parallel-group study. SETTING: Six teaching hospitals in Spain. PATIENTS: 55 patients with an apnea-hypopnea index of 30 or greater who did not have daytime sleepiness (Epworth Sleepiness Scale score /=5.7 mg/dL) (n = 13) but did not require immediate dialysis, patient and renal survival were 83% and 82% at 1 year and 62% and 69% at last follow-up. In patients who presented with dialysis-dependent renal failure (n = 39), patient and renal survival were 65% and 8% at 1 year and 36% and 5% at last follow-up. All patients who required immediate dialysis and had 100% crescents on renal biopsy remained dialysis dependent. CONCLUSIONS: Patients with the Goodpasture syndrome and severe renal failure should be considered for urgent immunosuppression therapy, including plasma exchange, to maximize the chance of renal recovery. Patients needing immediate dialysis are less likely to recover. PMID- 11388817 TI - The incidence of unrecognized myocardial infarction in women with coronary heart disease. AB - BACKGROUND: Several cohort studies in populations without coronary heart disease have demonstrated that up to 40% of incident myocardial infarctions are clinically unrecognized. OBJECTIVE: To determine the incidence of unrecognized myocardial infarction in women with coronary heart disease in the Heart and Estrogen/progestin Replacement Study (HERS). DESIGN: Randomized, double-blind, placebo-controlled trial of conjugated estrogens plus medroxyprogesterone or identical placebo with 4.1 years of follow-up. SETTINGS: Outpatient and community settings at 20 U.S. clinical centers. PATIENTS: 2763 postmenopausal women younger than 80 years of age with coronary heart disease and an intact uterus. MEASUREMENTS: Annual electrocardiograms were obtained for all participants during follow-up (mean, 4.1 years) and were evaluated by using the NOVACODE computer algorithm and visual confirmation. A total of 13 715 electrocardiograms were obtained. Suspected unrecognized myocardial infarctions were investigated by comparing a participant's previous surveillance electrocardiograms with the electrocardiograms obtained from all of her intervening hospitalizations. Characteristics of patients with recognized and unrecognized myocardial infarction were compared. RESULTS: Among the 256 nonfatal myocardial infarctions, 11 were unrecognized (4.3% [95% CI, 2.2% to 7.6%]). Seven occurred in women assigned to placebo and 4 occurred in women assigned to hormone therapy (P > 0.2). Women with unrecognized myocardial infarction were less likely to have diabetes mellitus or previous angina and were more likely to have had previous bypass surgery compared with women who had clinically recognized myocardial infarction. CONCLUSION: The incidence of unrecognized myocardial infarction in women with coronary disease was far lower than that observed in previous studies of populations without coronary heart disease. PMID- 11388818 TI - Diagnosis and care of patients with anorexia nervosa in primary care settings. AB - Anorexia nervosa is a psychiatric disorder characterized by abnormal eating behaviors that results in weight loss and has serious potential medical consequences. Most of these complications are readily treatable if diagnosed and attended to early in the course of the illness. In caring for patients with anorexia nervosa, the primary care physician has several critical roles. Because patients deny the severity of their illness, they delay seeking psychiatric treatment. The primary care physician must be skilled in recognizing this disorder, as well as in diagnosing and effectively treating the medical complications while educating the patient about them. The primary care physician is also involved with arranging and coordinating a comprehensive and multidisciplinary program, including dietary and mental health treatment. The multidisciplinary team is responsible for ensuring safe weight restoration and a judicious refeeding treatment plan. In addition to establishing the diagnosis and treating the multiple medical complications associated with anorexia nervosa, the primary care physician plays a central role in maintaining continuity of care despite the fact that successful care may require a variety of treatment settings. Factors that foster good prognoses for this increasingly common and often protracted eating disorder include early diagnosis and skilled medical intervention to prevent the inexorable physical decline that marked weight loss can cause. PMID- 11388819 TI - Pharmaceutical price controls and patient welfare. AB - Price controls could have a substantial negative effect on pharmaceutical research and development. Extensive research is required before the development costs of a new drug or its benefits are known; most new drug development projects fail, sometimes after substantial financial and time costs. These conditions pose intractable practical problems for the operation of price controls, which cannot rest on objective, predictable standards such as the benefits or costs of individual drugs. In the absence of objective standards, pressure from health care providers and others would create powerful incentives for price regulators to decrease drug prices toward marginal costs of production and distribution, well below levels sufficient to reward innovative research. This downwardly biased price-setting mechanism would apply with particular force to the few successful projects that yield innovative drugs, whose prices would not be set by regulatory authorities until after research expenditures have been incurred and the new drugs are ready to enter the market. Manufacturers will expect price controls to reduce the potential payoffs from breakthrough drugs. This expectation would substantially reduce the incentives to pursue innovative research, as is evident in advanced economies in which price controls are now in force. Once established, price controls for pharmaceuticals, like those for medical services in the Medicare system, would also tend toward complexity and entrenchment of vested interests and could easily become permanent regardless of the harm they cause to patients. PMID- 11388821 TI - The heartbreak of drug pricing. PMID- 11388820 TI - Who should get treated for sleep apnea? PMID- 11388823 TI - Doctor on D-Day. PMID- 11388824 TI - Cyclooxygenase-2 inhibition and renal function. PMID- 11388825 TI - Cyclooxygenase-2 inhibition and renal function. PMID- 11388827 TI - "Tell me about yourself": The patient-centered interview. PMID- 11388830 TI - Paesslerins A and B: novel tricyclic sesquiterpenoids from the soft coral Alcyonium paessleri. AB - [structure: see text] In the course of our search of new bioactive metabolites from marine invertebrates, paesslerins A and B, sesquiterpenoids with an unprecedented tricyclic skeleton, were isolated from the subAntarctic soft coral Alcyonium paessleri collected at a depth of 200 m near the South Georgia islands, and their structures were elucidated by spectroscopic techniques. These compounds show moderate cytotoxicity in preliminary assays. PMID- 11388831 TI - Synthesis, electronic spectra, and crystal structural properties of fluorinated [3(3)](1,3,5)cyclophanes. AB - [structure: see text] Trifluoro- and hexafluoro[3(3)](1,3,5)cyclophanes 3 and 4 were synthesized with TosMIC coupling as a key reaction. The pi-pi absorption bands show blue shifts as the number of fluorine atoms is increased. In the crystalline state, characteristic stacking with the fluorinated benzene rings facing each other is observed in both cases. PMID- 11388832 TI - Simple, tunable aziridination catalysts based on poly(pyrazolyl)borate-copper complexes. AB - [reaction: see text] The in situ generation of a copper-poly(pyrazolyl)borate complex from copper chloride and a sodium poly(pyrazolyl)borate salt results in a catalyst that is effective for the aziridination of olefins. A significant influence of the combination of the starting copper oxidation state and the hapticity of the poly(pyrazolyl)borate ligand on the efficiency of the reaction has been observed. PMID- 11388833 TI - Total synthesis of himandravine. AB - [reaction: see text] The first total synthesis of (+)-himandravine (1) is described, starting from (2S,6S)-cis-2-formyl-6-methyl-N-Boc-piperidine (8) in 11 linear steps and 17% overall yield. The key step involves a highly diastereoselective intramolecular Diels-Alder reaction of the key intermediate 5 that contains the entire latent carbon framework and functional group substitution of himandravine. PMID- 11388834 TI - The first tetrathiafulvalene- sigma-polynitrofluorene diads: low HOMO-LUMO gap, amphoteric redox behavior, and charge transfer properties. AB - [structure: see text] The synthesis, solution redox behavior, EPR, and intramolecular charge transfer properties of novel donor-acceptor diads of TTF sigma-A type (TTF = substituted tetrathiafulvalene, sigma = saturated spacer, A = polynitrofluorene acceptor) are reported. The HOMO-LUMO gap for compound 6 is as small as 0.3 eV, and spectroelectrochemical experiments reveal its electrochromic behavior in the near-IR region. PMID- 11388835 TI - Synthesis of ortho substituted arylboronic esters by in situ trapping of unstable lithio intermediates. AB - [reaction: see text] Ortho lithiation-in situ boration using lithium 2,2,6,6 tetramethylpiperidide (LTMP) in combination with triisopropylborate (B(OiPr)(3)) is a highly efficient and experimentally straightforward process for the preparation of ortho substituted arylboronic esters. The mild reaction conditions allow the presence of functionalities such as ester or cyano groups or halogen substituents that are usually not compatible with the conditions used in directed ortho metalation of arenes. The arylboronic esters underwent Suzuki-type cross coupling with a range of aryl halides, furnishing biaryls in 53-94% yield. PMID- 11388836 TI - Rearrangement kinetics of spiropentylchlorocarbene, [3]triangulylchlorocarbene, and related species. AB - [structure: see text] Absolute rate constants were measured for the 1,2-CH(2), 1,2-cyclopropyl, and 1,2-CMe(2) rearrangements of spiropentylchlorocarbene (5), [3]triangulylchlorocarbene (6), and tetramethylcyclopropylchlorocarbene (7). The factors responsible for the observed relative migratory aptitudes (cyclopropyl > CH(2) > CMe(2)) were analyzed with the aid of electronic structure calculations. PMID- 11388837 TI - Investigations into the production and interconversion of phomoidrides A-D. AB - [structure: see text] Fermentation of ATCC 74256 led to the isolation and identification of C7 epimers of phomoidride A (CP-225,917) and phomoidride B (CP 263,114). We suggest the names phomoidrides C and D for these new fermentation products. Studies on the effect of pH on the distribution of phomoidrides A-D suggest phomoidride B (CP-263,114) is the first-formed secondary metabolite and the source of the remaining three phomoidrides. PMID- 11388838 TI - Synthetic studies of antitumor natural products superstolides A and B. Construction of C20-C26 fragment of superstolide A. AB - [structure: see text] The C20-C26 portion of the antitumor macrolide superstolide A was synthesized by employing Brown's asymmetric crotylboronate methodology. PMID- 11388839 TI - Chemoselective cross-metathesis reaction. Application to the synthesis of the C1 C14 fragment of amphidinol 3. AB - [reaction: see text] An efficient synthesis of the C1-C14 fragment of amphidinol 3 is described. The synthesis is based on chemoselective cross-metathesis reactions and enantioselective allyltitanations. PMID- 11388840 TI - Oxidative cleavage of a cyclobutane pyrimidine dimer by photochemically generated nitrate radicals (no(3)*). AB - [reaction: see text] Photochemically generated nitrate radicals (NO(3)(*)) cleave the stereoisomeric N,N-dimethyl-substituted uracil cyclobutane dimers 1a-d into the monomeric uracil derivative 2 as the major reaction pathway. A preferred splitting of the syn dimers 1a,b was observed. The reaction is expected to proceed through initial one-electron oxidation with formation of an intermediate cyclobutane radical cation 11. In addition to cycloreversion, competing reaction steps of 11, which lead to the observed byproducts, are suggested. PMID- 11388841 TI - Chiral helicity induced by hydrogen bonding and chirality of podand histidyl moieties. AB - [structure: see text] The single-crystal X-ray structure determination of N,N' bis[(S)-(+)-1-methoxycarbonyl-2-(4-imidazolyl)ethyl]-2,6-pyridinedicarboxamide (L BHisPA) and the D-isomer (D-BHisPA) derived from the corresponding chiral histidine revealed a left- and right-handed helical conformation, respectively, through intramolecular hydrogen bonding and chirality of the podand histidyl moieties. Furthermore, each helical molecule is connected by continuous intermolecular hydrogen bonds to afford a left- or right-handed helical assembly, respectively, in the crystal packing. PMID- 11388843 TI - Novel alkali cation chemosensors based on n-9-anthrylaza-crown ethers. AB - [structure: see text] A novel alkali cation selective probe is described which exhibits unique ICT and binding properties, allowing the ratiometric titration of sodium cations in the presence of other alkali metal ions. PMID- 11388842 TI - Monitoring solid-phase glycoside synthesis with (19)F NMR spectroscopy. AB - [structure: see text] A simple and efficient method for monitoring and optimizing carbohydrate synthesis on polymeric support by using (19)F NMR spectroscopy is described. The method relies on the use of fluorinated variants of protective groups that are in common use in oligosaccharide synthesis. PMID- 11388844 TI - Solid phase synthesis of carbocyclic l-2'-deoxynucleosides. AB - [reaction: see text] Carbocyclic L-2'deoxynucleosides 17 were synthesized on solid phase in four steps from the appropriately protected intermedate 11. The Mitsunobu reaction was used as a condensation method between the carbocyclic moiety and heterocyclic bases. The regioselectivity of the carbocyclic nucleosides was compared between the solid and solution phase syntheses. PMID- 11388845 TI - Asymmetric intramolecular C_H insertions of aryldiazoacetates. AB - [reaction: see text] The enantioselectivity of Rh(2)(S-DOSP)(4) catalyzed C-H insertion of aryldiazoacetates is very dependent on the site of the C-H insertion. The highest enantioselectivity is obtained for insertion into methine C-H bonds. PMID- 11388846 TI - Isolation, structure determination, and synthesis of neodysiherbaine A, a new excitatory amino acid from a marine sponge. AB - [structure: see text] A new excitatory amino acid, neodysiherbaine A (2), was isolated as a minor constituent of the aqueous extract from the marine sponge Dysidea herbacea. The structure was deduced by spectroscopic methods and established unambiguously by the total synthesis. The present synthesis, including as a key step cross-coupling of the 6/5-bicyclic core with an amino acid residue, is useful in constructing its structural analogues. PMID- 11388848 TI - Direct mono-N-methylation of solid-supported amino acids: a useful application of the Matteson rearrangement of alpha-aminoalkylboronic esters. AB - [reaction: see text] A novel solid-phase method for the mono-N-methylation of resin-supported amino acids was developed on the basis of Matteson's 1,2-carbon to-nitrogen migration of boron in alpha-aminoalkylboronic esters. Amino acids supported on either Wang resin or the highly acid-sensitive SASRIN resin can be methylated by reaction with pinacol chloromethylboronic ester, followed by rearrangement of the resulting aminomethylboronate and subsequent cleavage of the boronate group. This direct method requires only a simple and expedient oxidative resin wash to repair overalkylated sites. PMID- 11388847 TI - Access to [6.4.0]carbocyclic systems by tandem metathesis of dienynes. A step toward the synthesis of a PreD3-D3 transition state analogue. AB - [reaction: see text] A new approach to the synthesis of linearly fused 6-8-6 tricarbocyclic systems, tandem ring-closing metathesis of dienynes, allows access to compounds with a carbon framework analogous to the proposed transition state of the isomerization of previtamin D(3) to vitamin D(3). PMID- 11388849 TI - A Nozaki-Hiyama-Kishi Ni(II)/Cr(II) coupling approach to the phomactins. AB - [reaction: see text] Through a unified synthetic strategy, appropriately functionalized bicyclic starting materials can be elaborated via Ni(II)/Cr(II) macrocylization to [9.3.1] bicycles. Elaboration of these core structures allows access to phomactin C/D analogues and establishes the first synthetic approach to phomactin A affording an intact octahydrochromene/macrocyclic ring system. PMID- 11388850 TI - A novel palladium-catalyzed intramolecular redox reaction. AB - [reaction: see text] A new type of palladium-catalyzed redox reaction is described, forming enones from 2-(2-bromobenzyl)-ketones with an overall loss of HBr. The scope and limitations of the reaction are demonstrated by a series of cyclic and acyclic substrates. The mechanism most probably involves the formation of an intramolecular arylpalladium enolate and is related to the oxidation of silyl enol ethers with palladium acetate. PMID- 11388851 TI - One-pot, catalytic, asymmetric synthesis of polypropionates. AB - [reaction: see text] The opening of methylketene dimer, followed by aldol reactions of the resulting enolate, provides a convenient access to syn,syn dipropionate aldol adducts of a variety of aldehydes. These aldol adducts are useful precursors in the synthesis of complex polypropionates. PMID- 11388852 TI - Water-accelerated tandem Claisen rearrangement-catalytic asymmetric carboalumination. AB - [reaction: see text] The addition of stoichiometric quantities of water accelerates both the trimethylaluminum-mediated aromatic Claisen reaction and the chiral zirconocene-catalyzed asymmetric carboalumination of terminal alkenes. The two reactions occur in a tandem sequence resulting in the selective formation of two new C-C and one C-O bond after oxidative quench of the intermediate trialkylalane. PMID- 11388854 TI - Highly efficient Heck reactions of aryl bromides with n-butyl acrylate mediated by a palladium/phosphine-imidazolium salt system. AB - [reaction: see text] A new phosphine-imidazolium salt, L.HBr (1, L = (1 ethylenediphenylphosphino-3-(mesityl))imidazol-2-ylidene), has been prepared. A combination of 0.5 mol % of Pd(dba)(2) and 0.5 mol % of L.HBr in the presence of 2 equiv of Cs(2)CO(3) as base has proven to be highly efficient in the Heck coupling reactions of aryl bromides (from electron-deficient to electron-rich aryl bromides) with n-butyl acrylate. PMID- 11388853 TI - Biocatalysis in ionic liquids: markedly enhanced enantioselectivity of lipase. AB - [reaction: see text] Lipase-catalyzed transesterifications in ionic liquids proceeded with markedly enhanced enantioselectivity. It was observed that lipases were up to 25 times more enantioselective in ionic liquids than in conventional organic solvents. PMID- 11388855 TI - Asymmetric alkylation of tert-butyl glycinate Schiff base with chiral quaternary ammonium salt under micellar conditions. AB - [reaction: see text] The asymmetric alkylation of the tert-butyl glycinate benzophenone Schiff base 1 with various arylmethyl bromides catalyzed by O-allyl N-(9-anthracenylmethyl)cinchonidinium bromide (2) proceeded smoothly under micellar conditions (5 equiv of 1 M KOH and 0.4 equiv of Triton X-100) to give the alkylated products in good yields and with good enantioselectivity (72-85% ee), depending on the electrophiles. PMID- 11388856 TI - A simple preparation of ketones. N-protected alpha-amino ketones from alpha-amino acids. AB - [reaction: see text] Carboxylic acids and amino acids are readily converted, under mild conditions, into the corresponding activated esters, which are reacted with Grignard/CuI reagent to give the corresponding ketones in nearly quantitative yields. The compounds were recovered substantially pure from the reaction mixtures. PMID- 11388857 TI - Basicity of some organic superbases in acetonitrile. AB - [structure: see text] It is shown that the pK(a) values of strong neutral organic (super)bases in acetonitrile are well described by the density functional theory (DFT) employing the isodensity polarization continuum model (IPCM) for treating solvent-solute interactions. High pK(a) values are predicted for two model compounds, and their synthesis is strongly recommended. PMID- 11388858 TI - Base-induced heterochiral dimerization of an oxiranyl carbaldimine: stereoselective synthesis of a highly functionalized aziridine. AB - [reaction: see text] Treatment of the oxiranyl carbaldimine with base (LDA or LDA/KOtBu) leads in an one-step procedure to the polyfunctionalized aziridine. This highly diastereoselective reaction is explained by a new type of an Aza Darzens reaction, in which one enantiomer of the starting material is deprotonated to form an oxiranyl anion, which attacks the imine carbon atom of the other enantiomer (mutual kinetic resolution by double diastereofacial selection). PMID- 11388859 TI - Model systems for flavoenzyme activity. Control of flavin recognition via specific electrostatic interactions. AB - [structure: see text] A model system has been used to study the interactions of dipole-containing aromatic systems with oxidized and reduced flavin. Ab initio computational and experimental studies show that dipole orientation within the host is a critical determinant for recognition and redox behavior of the flavin guest. PMID- 11388860 TI - Enantioselective strategy to the spirocyclic core of Palau'amine and related bisguanidine marine alkaloids. AB - [structure: see text] An enantioselective strategy to the spirocyclic core found in the oroidin-derived family of bisguanidine marine alkaloids has been devised, premised on a biosynthetic proposal. Herein, we describe the successful implementation of this strategy, which entails a Diels-Alder reaction and a chlorination/ring contraction sequence that delivers the fully functionalized spirocyclic core. In this initial report, an intermolecular chlorination delivers a cyclopentane that is epimeric at C17 relative to the palau'amines and epimeric at C11 relative to the axinellamines. PMID- 11388861 TI - Direct catalytic enantio- and diastereoselective aldol reaction using a Zn-Zn linked-BINOL complex: a practical synthesis of syn-1,2-diols. AB - [reaction: see text] The direct catalytic enantio- and diastereoselective aldol reaction with 2-hydroxy-2'-methoxyacetophenone proceeded smoothly using as little as 1 mol % of a dinuclear zinc catalyst, Zn-Zn-linked-BINOL complex 2, to afford alpha,beta-dihydroxy ketones in a highly syn-selective manner (up to syn/anti 97/3) and in excellent yields (up to 95%) and ees (up to 99%). Efficient transformations of the alpha,beta-dihydroxy ketone into an alpha,beta-dihydroxy ester and an alpha,beta-dihydroxy amide via regioselective rearrangements are also described. PMID- 11388862 TI - Novel bromotyrosine alkaloids: inhibitors of mycothiol S-conjugate amidase. AB - [structure: see text] The novel alkaloids 1 and 4 were isolated from an Australian non-verongid sponge, Oceanapia sp. Compound 1 contains an unprecedented imidazolyl-quinolinone substructure attached to a bromotyrosine derived spiro-isoxazoline. Three other known alkaloids were isolated in addition to 1 and 4 and together represent the first examples of inhibitors of a new mycobacterial enzyme mycothiol S-conjugate amidase (MCA). PMID- 11388863 TI - Synthesis of C-aryl and C-alkyl glycosides using glycosyl phosphates. AB - [reaction: see text] Mannosyl and glucosyl phosphate donors were successfully used in constructing C-aryl linkages common to many natural products via a Lewis acid induced Fries-like rearrangement. The rearrangement was stereo- and regiospecific, yielding only one C-glycoside product. C-Alkyl glycoside carbohydrate mimetics were generated by using silicon-derived C-nucleophiles and glycosyl phosphates. PMID- 11388864 TI - Synthesis and determination of absolute configuration of the bicyclic guanidine core of batzelladine A. AB - [reaction: see text] The synthesis of a selectively protected form of the bicyclic guanidine fragment of batzelladine A from L-aspartic acid is reported, thereby establishing the absolute configuration of the bicyclic guanidine ring system within the natural product. PMID- 11388867 TI - Progress toward the total synthesis of ingenol: construction of the complete carbocyclic skeleton. AB - [reaction: see text] The complete carbocyclic skeleton of ingenol is assembled via a route that employs ring-closing metathesis (RCM) to close the strained "inside-outside" BC ring system (i.e., 24 --> 25). PMID- 11388869 TI - Synthesis of protected L-4-[sulfono(difluoromethyl)phenylalanine and its incorporation into a peptide. AB - [reaction: see text] A protected form of L-4 [sulfono(difluoromethyl)]phenylalanine (F(2)Smp), a novel non-hydrolyzable phospho- and sulfotyrosine mimetic, was synthesized via electrophilic fluorination of a benzylic sulfonate followed by a Pd-catalyzed cross-coupling reaction between the fluorinated sulfonate and the zincate of protected iodoalanine. F(2)Smp was incorporated into a peptide using solid-phase peptide synthesis techniques. PMID- 11388870 TI - Efficient asymmetric synthesis of alpha-trifluoromethyl-substituted primary amines via nucleophilic 1,2-addition to trifluoroacetaldehyde SAMP- or RAMP hydrazone. AB - [reaction: see text] An efficient asymmetric synthesis of alpha-trifluoromethyl substituted primary amines via nucleophilic 1,2-addition of alkyllithium reagents to trifluoroacetaldehyde SAMP- or RAMP-hydrazone followed by benzoylation and SmI(2)-promoted nitrogen-nitrogen single bond cleavage is described. PMID- 11388871 TI - Solid-state photochromism and photoreactivity of o- and p-anisaldehydes. Remarkable stabilization of o-xylylenols. AB - [reaction: see text] All of the crystalline o-anisaldehyde derivatives 1 and 3 change to brick-red color upon brief exposure to UV-vis radiation. The red color, attributed to (E)-xylylenol, is remarkably persistent for hours (ca. 10 h) in the case of 1c; such a long lifetime for the reactive o-xylylenols is unprecedented. In contrast, the p-anisaldehydes 2 undergo cyclization. Solid-state photolysis of 2b affords the benzocyclobutenol 7b regioselectively, which is not accessible from solution-phase photolysis. PMID- 11388873 TI - A mild and chemoselective method for the reduction of conjugated isoxazolines to beta-hydroxy ketones. AB - A new procedure for the selective reduction of conjugated Delta(2)-isoxazolines to the corresponding unsaturated beta-hydroxy ketones is described. The use of SmI(2) as the reducing agent and B(OH)(3) to hydrolyze the resulting imine results in a mild, convenient, and chemoselective protocol for this otherwise difficult transformation and complements existing methodology for the preparation of beta-hydroxy ketones via nitrile oxides. PMID- 11388878 TI - Re: Surgery for fully accommodative esotropia. PMID- 11388879 TI - Editorial: The end of "fix, follow and maintain"; stereopsis in seniors; lorazepam for accommodative esotropia?; surgery for congenital fibrosis of the inferior rectus. PMID- 11388880 TI - Poor correlation between "fix-follow-maintain" monocular/binocular fixation pattern evaluation and presence of functional amblyopia. AB - PURPOSE: To assess the reliability of visual screening with fix- follow-maintain (FFM) method in early detection of amblyopia in children who are not able to cooperate in testing with Allen Figures or Snellen E-chart. METHODS: Followup records of 89 patients that were examined in our pediatric ophthalmology department between May 1996 and May 1999 were evaluated retrospectively. The visual acuities that were measured with FFM method at the first visit were compared with the visual acuity levels that were measured with the Snellen E chart at later followup visits. Functional amblyopia was defined as a difference of 2 or more Snellen lines between the two eyes. RESULTS: Amblyopia was later definitively diagnosed on the E-chart in only 17 (32.6%) of the 52 eyes that were estimated to have a significantly lower visual acuity than the fellow eye with the FFM method. Sensitivity of the FFM method in diagnosing amblyopia was found to be 53.1% and the specificity was 38.5%. CONCLUSIONS: The use of the FFM monocular/binocular fixation pattern to determine visual acuity and the possible presence of amblyopia is so insensitive, and so unspecific, that its reliability is very low and the results of such testing are therefore virtually useless, if not medically hazardous. There is, therefore, a need to make more universally available more sophisticated tests of vision (preferential looking, VEP); where they are not available, further emphasis should be placed on Snellen equivalent vision testing as with Allen Cards, Tumbling E, HOTV Test, Lea Figures etc. so as to obtain an accurate direct visual acuity at as young an age as possible. Amblyopia treatment should not be initiated solely on the basis of FFM testing. PMID- 11388881 TI - Assessment of adult stereopsis using the Lang 1 Stereotest: a pilot study. AB - BACKGROUND AND PURPOSE: To assess the use of the Lang 1 Stereotest as a vision screening test for adults, for which little is known or reported. METHOD: The Lang 1 Stereotest was administered to 292 consecutive participants of the Visual Impairment Project (VIP) five year followup study, which is a population based study of eye disease in Melbourne, Australia. 56.9% were female. The mean age was 59.4 years, range 44-90 years. A "positive" stereoscopic response was recorded where the stereoscopic target image was correctly named; a "partial positive" response where depth was appreciated but the shape could not be named; and a "negative" response where there was no appreciation of a stereo effect. The responses were further categorized so that the test was either "passed" or "failed". A "pass" score was 3/3 positive responses; 3/3 partial positive responses or 2/3 positive and/or partial positive responses where the negative response was at the 550" of arc stereoacuity level. "Failure" was 3/3 negative responses and 2/3 negative responses where the positive or partial positive response as at the 1200" level. RESULTS: Of the 292 participants tested, 19 (6.5%) participants "failed" the test. In addition to strabismus and anisometropia, failure was associated significantly (p<0.001) with reduced distance visual acuity (<6/12) irrespective of the ocular morbidity. CONCLUSION: The Lang 1 Stereotest correctly identified people with vision defects associated with reduced stereopsis. This suggests the Lang 1 Stereotest is an appropriate for the vision screening of adults as it is for children. PMID- 11388882 TI - Effects of lorazepam on vision and oculomotor balance. AB - BACKGROUND: Previous studies have shown an effect of the tranquilizer lorazepam on visual perception. We explored the effects of the drug on binocular vision, visual acuity and accommodation. SUBJECTS AND METHODS: Twenty-four paid healthy volunteers (13 women, 11 men) were recruited from the University of Strasbourg (mean age: 23.6 years, mean weight: 66.8 Kg). They were randomly assigned to one of two parallel groups of 12 subjects each (a placebo group and a lorazepam 0.038 mg/kg group). Visual acuity was measured for each eye separately (Snellen chart and Parinaud scale). Binocular vision was studied using the cover tests, measurement of the fusional amplitudes (with Berens prisms), and the Duane Scale Test (near point rule) measuring convergence and/or accommodation in centimeters or diopters as a function of age. RESULTS: Regarding vision, there was no lorazepam effect, at either 33 cm or 5 m. An esophoria was observed after the intake of lorazepam (0Delta before intake and 2.8Delta after intake, p=0.001). Both fusional convergence and fusional divergence amplitudes decreased by lorazepam, (p=0.008, and p=0.002). Lorazepam also impaired the near point of convergence but did not affect accommodation. CONCLUSION: A single dose of lorazepam induces an esophoric oculomotor imbalance and impaired fusional convergence and divergence amplitudes without impairing visual acuity or accommodation. PMID- 11388883 TI - Outcome of surgery for congenital fibrosis of the inferior rectus muscle. AB - PURPOSE: To report clinical findings and surgical outcome in a large series of patients with fibrosis of the inferior rectus muscle. MATERIALS AND METHODS: Subject Cases: A total of 17 cases were diagnosed with unilateral fibrosis of the inferior rectus muscle during the past 27 years at our institution. They were aged from 5 months to 17 years, with 15 cases under 10 years of age. No differences were present regarding the laterality or gender. FINDINGS: All the cases showed hypotropia with restricted eye elevation. Forced duction test showed resistance to upward eye movement. A horizontal deviation in primary eye position was also present in 10 cases (59%). The affected eye was amblyopic in the majority of cases. Binocular vision was absent in 15 of the 17 cases. RESULTS: Surgical Outcome: All the cases received either recession or free tenotomy of the inferior rectus muscle. Resection of the ipsilateral superior rectus muscle was additionally performed to correct residual hypotropia. Fibrosis of the inferior rectus was present as intraoperative finding in all the 17 cases. Hypotropia disappeared in 10 cases and decreased in 7 cases. Restoration of satisfactory binocular alignment was obtained in all the 17 cases. CONCLUSION: Recession of the inferior rectus muscle was effective treatment for fibrosis of the inferior rectus. Additional resection of the ipsilateral superior rectus muscle was useful to correct residual hypotropia. Free tenotomy is not recommended. PMID- 11388884 TI - A case of esotropia associated with gaze limitation after 9 previous surgical procedures. PMID- 11388885 TI - Anomalous medial rectus muscle insertion in a child with craniosynostosis. AB - INTRODUCTION: Oblique and vertical rectus muscle anomalies have been commonly reported in patients with craniofacial syndromes, while horizontal rectus muscle anomalies have been uncommonly reported. METHODS: Case report of a child with Crouzon's Syndrome who was found to have an anomalous medial rectus muscle insertion at surgery. RESULTS: A bifid left medial rectus muscle insertion was found at surgery, requiring a small modification of the planned surgical procedure. CONCLUSION: Anomalies of extraocular muscles may be present in patients with craniofacial syndromes and strabismus surgeons should be prepared to modify their surgical plan when anomalous extraocular muscles are found. PMID- 11388887 TI - Reversible renal impairment induced by treatment with the angiotensin II receptor antagonist candesartan in a patient with bilateral renal artery stenosis. AB - BACKGROUND: It is well established that ACE-inhibitors should be avoided in patients with renal artery stenosis. In recent years it has also been recommended that caution should be demonstrated when angiotensin II blockers are used in the same type of patients but the evidence is based only on few cases. RESULTS: We describe a case where use of the angiotensin II antagonist candesartan (Atacand) induced renal failure in a patient with bilateral renal artery stenosis. The course of the case is enlighted by results from sequential renography, selective renal vein catheterisation for measurement of renin, and angiographic findings. CONCLUSIONS: In patients with renal artery stenosis the angiotensin II antagonist candesartan should be avoided. PMID- 11388888 TI - Encouraging good antimicrobial prescribing practice: a review of antibiotic prescribing policies used in the South East Region of England. AB - BACKGROUND: Good prescribing practice has an important part to play in the fight against antimicrobial resistance. Whilst it was perceived that most hospitals and Health Authorities possessed an antibiotic policy, a review of antibiotic policies was conducted to gain an understanding of the extent, quality and usefulness of these policies. METHODS: Letters were sent to pharmacists in hospitals and health authorities in across the South East region of the National Health Service Executive (NHSE) requesting antibiotic policies. data were extracted from the policies to assess four areas; antibiotic specific, condition specific, patient specific issues and underpinning evidence. RESULTS: Of a possible 41 hospital trusts and 14 health authorities, 33 trusts and 9 health authorities (HAs) provided policies. Both trust and HA policies had a median publication date of 1998 (trust range 1993-99, HA 1994-99). Eleven policies were undated. The majority of policies had no supporting references for the statements made. All policies provided some details on specific antibiotics. Gentamicin and ciprofloxacin were the preferred aminoglycoside and quinolone respectively with cephalosporins being represented by cefuroxime or cefotaxime in trusts and cephradine or cephalexin in HAs. 26 trusts provided advice on surgical prophylaxis, 17 had meningococcal prophylaxis policies and 11 covered methicillin resistant Staphylococcus aureus (MRSA). There was little information for certain groups such as neonates or children, the pregnant or the elderly. CONCLUSION: There was considerable variation in content and quality across policies, a clear lack of an evidence base and a need to revise policies in line with current recommendations. PMID- 11388889 TI - Molecular cloning, functional characterization and genomic organization of four alternatively spliced isoforms of the human organic cation transporter 1 (hOCT1/SLC22A1). AB - In this study we report the cloning of four human OCT1 (hOCT1/SLC22A1) isoforms: a long form, hOCT1G/L554, and three shorter forms (hOCT1G/L506, hOCT1G483 and hOCT1G353). All four variants could be identified in the human glioma cell line SK-MG-1, whereas only two isoforms (hOCT1G/L554 and hOCT1G/L506) were found in human liver cDNA. The hOCT1G/L554 represents the full length hOCT1 since the sequence of this clone is more than 99% identical to previously cloned hOCT1 cDNAs. Elucidation of the gene structure of human OCT1 demonstrated that the other isolated isoforms are alternatively spliced variants. The hOCT1 gene consists of 7 exons and 6 introns. When stably expressed in human embryonic kidney (HEK293) cells, only the full length hOCT1 cDNA mediated decynium-22 (D22) sensitive uptake of tritiated 1-methyl-4-phenylpyridinium ([3H]-MPP+). PMID- 11388891 TI - Applications of the estimating equations theory to genetic epidemiology: a review. AB - Unlike monogenic diseases for which considerable progress has been made in past years, the identification of susceptibility genes involved in multifactorial diseases still poses numerous challenges, including the development of new statistical methodologies. Recently, several authors have advocated the use of the estimating equations (EE) approach as an alternative to standard maximum likelihood methods for analysing correlated data. Since most genetic studies rely on family data, the EE found a natural field of application in genetic epidemiology. The objective of this review is to give a brief description of the EE principles, and to outline its applications in the main areas of genetic epidemiology, including familial aggregation analysis, segregation analysis, linkage analysis and association studies. PMID- 11388892 TI - Deletion pattern in the dystrophin gene in Turks and a comparison with Europeans and Indians. AB - Patterns of dystrophin gene deletions in DMD/BMD patients were compared in four populations: Turks (n = 146 deletions), Europeans (n = 838), North Indians (n = 89), and Indians from all over India (n = 103). Statistical tests revealed that there are differences in the proportions of small deletions. In contrast, the distribution of deletion breakpoints and the frequencies of specific deletions commonly observed in the four populations are not significantly different. The variations strongly suggest that sequence differences exist in the introns, and the differences are in agreement with genetic distances among populations. The similarities suggest that some intronic sequences have been conserved and that those will trigger recurrent deletions, since it is unlikely that gene flow would disperse the deleted chromosomes, which vanish from the gene pool in a few generations. PMID- 11388898 TI - Codon and base biases after the initiation codon of the open reading frames in the Escherichia coli genome and their influence on the translation efficiency. AB - Nucleotide sequences around the boundaries of all open reading frames in the Escherichia coli whole genome were analyzed. Characteristic base biases were observed after the initiation codon and before the termination codon. We examined the effect of the base sequence after the initiation codon on the translation efficiency, by introducing mutations after the initiation codon of the E. coli dihydrofolate reductase (DHFR) gene, considering codon and base biases, and using in vitro and in vivo translation systems. In both assay systems, the two most frequent second codons, AAA and AAU, enhanced the translation efficiency compared with the wild type, whereas the effects of lower frequency codons were not significant. Experiments using 16S rRNA variants with mutations in the putative complementary sequence to the region downstream of the initiation codon showed that the translation efficiency of none of the DHFR mutants was affected. These results demonstrate that the statistically most frequent sequences for the second codon enhance translation efficiency, and this effect seems to be independent of base pairing between mRNA and 16S rRNA. PMID- 11388899 TI - Detergent-resistant membrane domains are required for mast cell activation but dispensable for tyrosine phosphorylation upon aggregation of the high affinity receptor for IgE. AB - Aggregation of the high affinity receptor for IgE (FceRI) on mast cells results in the rapid phosphorylation of tyrosines on the beta and gamma chains of the receptor by the Src family kinase Lyn, which initiates the signaling cascades leading to secretion of inflammatory mediators. The detergent-resistant membranes (DRMs) have been implicated in FcepsilonRI signaling because aggregated receptors emigrate to DRMs that are enriched in certain signaling components. We evaluated the role of DRMs in FcepsilonRI signaling by disruption of DRMs using a cholesterol-binding agent, methyl-beta-cyclodextrin (MBCD). While treatment of rat basophilic leukemia cells with MBCD inhibits degranulation and Ca(2+) mobilization upon aggregation of FcepsilonRI, MBCD hardly affects the aggregation induced tyrosine phosphorylation of FcepsilonRI as well as other signaling molecules such as phospholipase C-gamma1 (PLC-gamma1). MBCD delocalizes phosphatidylinositol 4,5-bisphosphate from DRMs, which may prevent MBCD-treated cells from producing inositol 1,4,5-trisphosphate by means of activated PLC gamma1. These data suggest an indispensable role for DRMs in the Ca(2+) response rather than tyrosine phosphorylation, and support a model of receptor phosphorylation in which aggregated FcepsilonRI is tyrosine phosphorylated outside DRMs by constitutively associated Src family kinase Lyn via a transphosphorylation mechanism. PMID- 11388900 TI - Quantum chemical study of agonist-receptor vibrational interactions for activation of the glutamate receptor. AB - To understand the mechanism of activation of a receptor by its agonist, the excitation and relaxation processes of the vibrational states of the receptor should be examined. As a first approach to this problem, we calculated the normal vibrational modes of agonists (glutamate and kainate) and an antagonist (6-cyano 7-nitroquinoxaline-2,3-dione: CNQX) of the glutamate receptor, and then investigated the vibrational interactions between kainate and the binding site of glutamate receptor subunit GluR2 by use of a semiempirical molecular orbital method (MOPAC2000-PM3). We found that two local vibrational modes of kainate, which were also observed in glutamate but not in CNQX, interacted through hydrogen bonds with the vibrational modes of GluR2: (i) the bending vibration of the amine group of kainate, interacting with the stretching vibration of the carboxyl group of Glu705 of GluR2, and (ii) the symmetric stretching vibration of the carboxyl group of kainate, interacting with the bending vibration of the guanidinium group of Arg485. We also found collective modes with low frequency at the binding site of GluR2 in the kainate-bound state. The vibrational energy supplied by an agonist may flow from the high-frequency local modes to the low frequency collective modes in a receptor, resulting in receptor activation. PMID- 11388901 TI - Isolation of NPC1-deficient Chinese hamster ovary cell mutants by gene trap mutagenesis. AB - Chinese hamster ovary cell mutants defective in the NPC1 gene (NPC1-trap) were generated by retrovirus-mediated gene trap mutagenesis from a parental cell line JP17 expressing an ecotropic retrovirus receptor. Insertion of the gene trap vector in the NPC1 gene and the absence of the gene product were verified by 5'RACE and immunological analyses, respectively. NPC1-trap cells showed intracellular accumulation of low-density lipoprotein (LDL)-derived cholesterol and had an increased level of unesterified cellular cholesterol. Cholesterol biosynthesis through the mevalonate pathway was upregulated in the mutant cells as assessed by [(14)C]acetate incorporation into cellular sterols. When JP17 cells were depleted of lipoproteins and then loaded with LDL, cell surface LDL receptors were promptly downregulated and the mature form of the sterol regulatory element-binding protein-1 disappeared from the nucleus. These responses to LDL were obviously retarded in NPC1-trap cells, suggesting an impaired response of the cholesterol-regulatory system to LDL. NPC1-trap cells will be a useful tool to study the regulation of cellular cholesterol homeostasis and the pathogenesis of Niemann-Pick disease type C. PMID- 11388902 TI - Myosin assembly critical for the enzyme activity of smooth muscle myosin phosphatase: effects of MgATP, ionic strength, and Mg(2+). AB - We suggested that an assembled form of phosphorylated myosin (P-myosin) might exhibit higher affinity for smooth muscle myosin phosphatase (SMMP) than dissociated P-myosin on the basis of the effect of MgATP [Sato and Ogawa (1999) J. Biochem. 126, 787-797]. To further deepen our understanding, we examined the SMMP activity and P-myosin assembly with various ionic strengths and Mg(2+) concentrations, with and without MgATP, all of which are well known to be critical for myosin assembly. The structure of myosin molecules was directly observed by electron microscopy using a rotary shadowing procedure, which was found to be consistent with the sedimentation assay. We found that the SMMP activity was always high when P-myosin was assembled. MgATP, which disassembled P myosin mostly into a folded conformation, in contrast, decreased the enzyme activity. We also found that glycerol had a dissociating action on P-myosin, primarily dissociating it into an extended conformation, resulting in reduced SMMP activity, and that increases in the ionic strength and Mg(2+) (>5 mM) inhibited SMMP. These results indicate that myosin assembly is essential for SMMP activity. PMID- 11388903 TI - Effect of cholesterol on the formation of an interdigitated gel phase in lysophosphatidylcholine and phosphatidylcholine binary mixtures. AB - We previously reported that 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) forms an interdigitated gel phase in the presence of 1-palmitoyl-sn-glycero-3 phosphocholine (16:0LPC) at concentrations below 30 mol%. In the present investigation, fluorescent probe 1,6-diphenyl-1,3,5-hexatriene (DPH), X-ray diffraction, and differential scanning calorimetry (DSC) were used to investigate the effect of cholesterol on the phase behavior of 16:0LPC/DPPC binary mixtures. At 25 degrees C, 30 mol% 16:0LPC significantly decreases the DPH fluorescence intensity during the transition of DPPC from the L(beta') phase to the L(betaI) phase. However, the addition of cholesterol to 16:0LPC/DPPC mixtures results in a substantial increase in fluorescence intensity. The changes in DPH fluorescence intensity reflect the probe's redistribution from an orientation parallel to the acyl chain to the center of the bilayer, suggesting a bilayer structure transition from interdigitation to noninterdigitation. The normal repeat period of small angle X-ray diffraction patterns can be restored and a reflection appears at 0.42 nm with a broad shoulder around 0.41 nm in wide angle X-ray diffraction patterns when 10 mol% cholesterol is incorporated into 30 mol% 16:0LPC/DPPC vesicles, indicating that the mixtures are in the gel phase (L(beta')). Moreover, DSC results demonstrate that 10 mol% cholesterol is sufficient to significantly decrease the main enthalpy, cooperativity and lipid chain melting of 30 mol% 16:0LPC/DPPC binary mixtures, which are L(betaI), indicating that the transition of the interdigitated phase is more sensitive to cholesterol than that of the noninterdigitated phase. Our data imply that the interdigitated gel phase induced by 16:0LPC is prevented in the presence of 10 mol% cholesterol, but unlike ethanol, an increasing concentration of 16:0LPC is not able to restore the interdigitation structure of the lipid mixtures. PMID- 11388904 TI - Frontal gel chromatographic analysis of the interaction of a protein with self associating ligands: aberrant saturation in the binding of flavins to bovine serum albumin. AB - Frontal gel chromatography is an accurate method to obtain the total free ligand concentration of a protein-ligand mixture in which ligands self-associate. The average number of bound ligands per protein molecule is obtained as a function of the total free ligand concentration. The method was applied to the interaction of bovine serum albumin with self-associating flavins. The binding curves for FMN and FAD leveled off at about 0.7 and 0.5, respectively. These data were simulated well by a binding model where flavins undergo isodesmic indefinite self association and the monomer alone binds to a single binding site of albumin. The isodesmic association constants of FMN and FAD were (1.7 +/- 0.1) x 10(2) and (2.2 +/- 0.3) x 10(2) M(-1), respectively. The binding constants of the monomer of FMN and FAD were (7.6 +/- 0.2) x 10(2) and (3.5 +/- 0.2) x 10(2) M(-1), respectively. FMN competitively inhibited the binding of FAD to albumin. The affinity to flavins was in the following order at pH 5.8: lumiflavin, FMN, riboflavin, and FAD. The SH modification and the binding of palmitate did not affect the FMN binding to bovine albumin at pH 5.8. As pH increased from 5.8 to 9.0, the affinity to FMN of bovine albumin decreased 3-fold, whereas that of human albumin increased about 80-fold. The present study clearly showed how isodesmic self-association of a ligand can cause apparent saturation of the interaction of a protein with the ligand at levels lower than 1. PMID- 11388905 TI - A theoretical study of electronic and structural states of neurotransmitters: gamma-aminobutyric acid and glutamic acid. AB - As a first approach to understanding the mechanism for the recognition of a ligand by its receptor, we first calculated the electronic and structural states of ionized gamma-aminobutyric acid (GABA) and ionized glutamic acid using the ab initio method with the 6-311++G (3df, 2pd) basis set. We paid special attention to the physicochemical characteristics of these molecules, such as the electric dipole moment, electrostatic potential, and electrostatic force. Even though GABA and glutamic acid are known to exert completely opposite influences in the mammalian brain by binding their specific receptors, the only difference in their chemical structures is that glutamic acid contains one more carboxyl group than GABA. As a result, we succeeded in showing that a difference of only one carboxyl group induces significant differences in the electronic and structural states between these molecules. These differences have a crucial influence on the electric dipole moments, the electrostatic potentials, and the electrostatic forces. The most remarkable finding of the present research is that the electrostatic potential formed by glutamic acid is composed of only negative parts, while that formed by GABA is separated into positive and negative parts. These results strongly suggest that GABA can approach either positively or negatively charged amino acids by adjusting its own orientation, while glutamic acid can approach only a positively charged binding site. PMID- 11388906 TI - Structural and functional changes in bovine pancreatic ribonuclease a by the replacement of Phe120 with other hydrophobic residues. AB - To clarify the specific role of Phe120 in bovine pancreatic ribonuclease A (RNase A), changes in the thermal stability and activity of F120L, F120A, F120G, and F120W were analyzed with respect to some thermodynamic terms, i.e., Gibbs free energy, enthalpy, and entropy. The structural destabilization of F120L, F120A, and F120G was due to a decrease in DeltaH(m) with a parallel decrease in amino acid volume at position 120, while the destabilization of F120W can be ascribed to an increase in DeltaS(m) accompanying an increase in DeltaH(m), showing that the size of Phe120 produces an optimum balance of conformational enthalpy and entropy for achieving the maximal structural stability. Moreover, the replacement of Phe120 affects activity. The increase in K(m) showed that the hydrophobicity and pi electron of Phe120 are important factors in substrate binding. The decrease in k(cat) was predicted to be due to positional changes of the side chains of His12 and/or His119. The positional changes were successfully detected by the rate of carboxymethylation by iodoacetate or bromoacetate, which correlated very well with decreases in activity, supporting the view that Phe120 also plays an important role in determining the position of His12 and/or His119 in order to achieve efficient catalysis. PMID- 11388907 TI - Molecular cloning and expression of endo-beta-N-acetylglucosaminidase D, which acts on the core structure of complex type asparagine-linked oligosaccharides. AB - Endo-beta-N-acetylglucosaminidase D (Endo D) produced by Streptococcus pneumoniae cleaves the di-N-acetylchitobiose structure in asparagine-linked oligosaccharides. The enzyme generally acts on complex type oligosaccharides after removal of external sugars by neuraminidase, beta-galactosidase, and beta-N acetylglucosaminidase. We cloned the gene encoding the enzyme and expressed it as a periplasm enzyme in Escherichia coli. The first 37 amino acids in the predicted sequence are removed in the mature enzyme, yielding a protein with a molecular mass of 178 kDa. The substrate specificity of the recombinant enzyme is indistinguishable from the enzyme produced by S. pneumoniae. Endo-beta-N acetylglucosaminidase A (Endo A) from Arthrobacter protophormiae, the molecular mass of which is 72 kDa, had 32% sequence identity to Endo D, starting from the N terminal sides of both enzymes, although Endo A hydrolyzes high-mannose-type oligosaccharides and does not hydrolyze complex type ones. Endo D is not related to endo-beta-N-acetylglucosaminidases H, F(1), F(2), or F(3), which share common structural motifs. Therefore, there are two distinct groups of endo-beta-N acetylglucosaminidases acting on asparagine-linked oligosaccharides. The C terminal region of Endo D shows homology to beta-galactosidase and beta-N acetylglucosaminidase from S. pneumoniae and has an LPXTG motif typical of surface-associated proteins of Gram-positive bacteria. It is possible that Endo D is located on the surface of the bacterium and, together with other glycosidases, is involved in virulence. PMID- 11388908 TI - Secretion of non-helical collagenous polypeptides of alpha1(IV) and alpha2(IV) chains upon depletion of ascorbate by cultured human cells. AB - Our previous report showed that human fetal lung fibroblasts secreted non disulfide-bonded, non-helical collagenous polypeptides of alpha1(IV) and alpha2(IV) chains depending on culture conditions [Connective Tissue (1999) 31, 161-168]. The secretion of non-helical collagenous polypeptides is unexpected from the current consensus that such polypeptides are not secreted under physiological conditions. The absence of interchain disulfide bonds among alpha1(IV) and alpha2(IV) chains was always correlated with the absence of triple helical structure of the type IV collagen. The finding corresponds with the fact that the interchain disulfide bonds are formed at or close to the completion of the type IV collagen triple-helix formation. The present report shows that ascorbate is the primary factor for the triple-helix formation of the type IV collagen. When human mesangial cells were cultured with ascorbate, only the triple-helical type IV collagen was secreted. However, when the cells were cultured without ascorbate, the non-helical alpha1(IV) and alpha2(IV) chains were secreted. Relative amounts of the secreted products were unchanged with or without ascorbate, suggesting that ascorbate is required for the step of the triple-helix formation. The ascorbate-dependency of the triple-helix formation of the type IV collagen was observed in all the human cells examined. The non helical alpha1(IV) chain produced by the ascorbate-free culture contained about 80% less hydroxyproline than the alpha1(IV) chain from the triple-helical type IV collagen. The evidence for the non-association of the non-helical alpha1(IV) and alpha2(IV) chains in the conditioned medium was obtained by an anti-alpha1(IV) antibody-coupled affinity column chromatography for the conditioned medium. Although all the non-helical alpha1(IV) chains were found in the bound fraction, all the non-helical alpha2(IV) chains were recovered in the flow-through fraction. The present findings suggest that ascorbate plays a key role in the trimerization step of three alpha chains and/or in the subsequent triple-helix formation of the type IV collagen. PMID- 11388909 TI - The down-regulation of glutathione peroxidase causes bovine luteal cell apoptosis during structural luteolysis. AB - Prostaglandin (PG) F(2)a is known to initiate luteal cell apoptosis in the bovine corpus luteum (CL) via its specific receptor (FP) on the luteal membrane by inducing intracellular Ca(2+) mobilization and the activation of PKC. In order to identify the signaling components involved in cell apoptosis, mRNA levels and activities of antioxidative enzymes were analyzed using bovine CL at different stages of the estrous cycle. Northern blot analysis revealed that the levels of two isozymes of superoxide dismutase (SOD), the Mn and Cu/Zn types, and catalase are highly enriched in the middle estrous phase, whereas glutathione peroxidase (GPx) levels gradually decrease as the estrous cycle progresses. The incubation of bovine luteal cells with H(2)O(2) and mercaptosuccinate (MS), a specific inhibitor of GPx, resulted in an increase in chromatin DNA condensation and apoptotic DNA fragmentation. Analyses of the enzymatic activities of GPx and catalase support the RNA data, indicating that H(2)O(2) produced due to the lack of GPx is a potent inducer of luteal cell apoptosis. PMID- 11388910 TI - Demonstration of the importance and usefulness of manipulating non-active-site residues in protein design. AB - Do non-active-site residues participate in protein function in a more direct way than just by holding the static framework of the protein molecule? If so, how important are they? As a model to answer these questions, ATB17, which is a mutant of aspartate aminotransferase created by directed evolution, is an ideal system because it shows a 10(6)-fold increase in the catalytic efficiency for valine but most of its 17 mutated residues are non-active-site residues. To analyze the roles of the mutations in the altered function, we divided the mutations into four groups, namely, three clusters and the remainder, based on their locations in the three-dimensional structure. Mutants with various combinations of the clusters were constructed and analyzed, and the data were interpreted in the context of the structure-function relationship of this enzyme. Each cluster shows characteristic effects: for example, one cluster appears to enhance the catalytic efficiency by fixing the conformation of the enzyme to that of the substrate-bound form. The effects of the clusters are largely additive and independent of each other. The present results illustrate how a protein function is dramatically modified by the accumulation of many seemingly inert mutations of non-active-site residues. PMID- 11388911 TI - A novel mixed valence form of Rhus vernicifera laccase and its reaction with dioxygen to give a peroxide intermediate bound to the trinuclear center. AB - Rhus vernicifera laccase, in a novel mixed valence state [T1oxT23red: type 1 Cu as Cu(II), and type 2 and 3 Cus as Cu(I)], was formed by reacting Cu(I) on the type 2 Cu-depleted laccase [T1oxT3red: type 1 Cu as Cu(II) and type 3 Cus as Cu(I)] under argon. Contrary to T1oxT3red, T1oxT23red was highly reactive with dioxygen, and gave the three transient bands at 340, 475, and 680 nm due to the two-electron reduced form of dioxygen [charge transfer bands from peroxide to Cu(II)]. The first order decays were highly dependent on pH, which led to the successful detection of the intermediate for ca. 2 h at pH 7.5. Another mixed valence derivative, T12oxT3red [type 1 and type 2 Cus as Cu(II), and type 3 Cus as Cu(I)] prepared through the action of Cu(II) on T1oxT3red was not reactive with dioxygen, but showed high enzyme activity as to the oxidation of N,N dimethyl-p-phenylenediamine. The whole reaction mechanism of the reduction of dioxygen by laccase was proposed based on the present results together with data for the former detection and characterization of the three-electron reduced form of dioxygen [Huang, H. et al. (1999) J. Biol. Chem. 274, 46, 32718-32724]. PMID- 11388912 TI - Enhancement of the thermostability of thermophilic bacterium PS-3 PPase on substitution of Ser-89 with carboxylic amino acids. AB - Serine 89 of the inorganic pyrophosphatase (PPase) subunit from thermophilic bacterium PS-3 (PS-3) was replaced with glycine, alanine, threonine, glutamic acid, or aspartic acid by the PCR-mutagenesis method with Mut-1 in order to determine the contribution of this serine residue to the thermostability and structural integrity of the enzyme molecule. S89G, S89A, and S89T showed reduced catalytic activity, whereas S89D and S89E showed increased enzyme activity. S89G, S89A, and S89T as well as the wild-type PPase were stable in the presence of 5 mM MgCl(2) at 70 degrees C for 1 h, but were inactivated rapidly with time at 80 degrees C. On the contrary, S89D and S89E were stable at 80 degrees C, showing more than 95% of the original activity after 1 h incubation. The wild-type PPase, S89D and S89E were each a hexamer before and after incubation at 80 degrees C for 1 h, while S89G and S89A comprised a mixture of a hexamer and a trimer both before and after incubation at 80 degrees C for 1 h. On the other hand, S89T was a mixture of a hexamer, a trimer and a monomer, and it was partially precipitated during heat treatment at 80 degrees C. The CD spectra of the recombinant enzymes in the far-ultraviolet region were the same as that of the wild-type PPase, whereas those of S89G, S89A, and S89T as well as the wild-type PPase were markedly different after heat treatment, although those of S89D and S89E did not change. The present study suggested that local small change(s) in the network of interactions among amino acid residues on replacement at position 89 led to the PS-3 PPase molecule being unable to form a hexamer from trimers or to dissociate into monomers in some cases without a significant change in the backbone conformation. It was also suggested that the partial disordering of the conformation of PS-3 PPase caused by heat depended on the degree of hydrophilicity in the vicinity of position 89. PMID- 11388914 TI - Protein anatomy: structure and function of peptide fragments corresponding to the secondary structure units of barnase. AB - Globular proteins can be decomposed into several modules or secondary structure units. It is useful to investigate the functions of such structural units in order to understand the folding units of proteins. In our previous work, barnase was divided into six peptide fragments corresponding to modules, and some of them were shown to have RNA-binding and RNase activity [Yanagawa, et al. (1993) J. Biol. Chem. 268, 5861-5865]. Barnase mutant proteins obtained by permutation of the structural units also had RNase activity [Tsuji, T. et al. (1999) J. Mol. Biol. 286, 1581-1596]. Here we investigated the structure and function of peptide fragments corresponding to secondary structure units of barnase. The results of circular dichroism spectroscopy indicated that some of the peptide fragments form helical structures in aqueous solutions containing over 30% 2,2,2 trifluoroethanol, and the S6 (94-110) peptide fragment is induced to form a beta sheet structure in the presence of RNA. The S6 peptide fragment forms aggregate complexes with RNA. Electron microscopic analysis showed that the aggregate complexes were comprised of filaments. These results indicate that not only modules but also secondary structure units dissected from a globular protein have functional and structure-forming capabilities. PMID- 11388913 TI - Caspase-independent cell death and mitochondrial disruptions observed in the Apaf1-deficient cells. AB - Apaf1 is a critical molecule in the mitochondria-dependent apoptotic pathway. Here we show that Apaf1-deficient embryonic fibroblasts died at a later phase of apoptotic induction, although these cells were resistant to various apoptotic stimulants at an early phase. Neither caspase 3 activation nor nuclear condensation was observed during this cell death of Apaf1-deficient cells. Electron microscopic examination revealed that death in response to apoptotic stimulation resembled necrosis in that nuclei were round and swollen with low electron density. Necrosis-like cell death was also observed in wild-type cells treated with z-VAD-fmk. Mitochondria were not only morphologically abnormal but functionally affected, since mitochondrial transmembrane potential (DeltaPsim) was lost even in cells with intact plasma membrane integrity. These mitochondrial alterations were also observed in the wild-type cells dying of apoptosis. Combined, these data suggest that cells without caspase activation, such as Apaf1 deficient cells or cells treated with caspase inhibitors, die of necrosis-like cell death with mitochondrial damage in response to "apoptotic stimulation." PMID- 11388915 TI - Cryoprotective effect of the serine-rich repetitive sequence in silk protein sericin. AB - The silk proteins, fibroin and sericin, are produced in the silk gland of Bombyx mori, and hydrophilic sericin envelops fibroin with successive sticky layers in the formation of a cocoon. To study the biological functions of sericin, we focused on the serine-rich sericin peptide consisting of 38 amino acids, which is a highly conserved and internally repetitive sequence of a sericin protein. The corresponding gene was chemically synthesized, and the PCR-amplified gene was ligated to oligomerize sericin peptide and fused at the amino terminus to a His tagged and proteolytic cleavage sequence in an inducible expression vector. When the dimers of sericin peptides were overexpressed in Escherichia coli, the transformants showed a prominent increase in cell viability after freezing in medium. Further, the purified dimeric sericin peptide from E. coli was found to be effective in protecting lactate dehydrogenase from denaturation caused by freeze-thaw. Both of these protective effects against freezing stress in cells and proteins were also observed with sericin hydrolysate. These results indicate that this unique sericin peptide, like sericin, has a high cryoprotective activity and will be valuable as a new biomaterial for industrial use. PMID- 11388916 TI - Dynamic diaschisis: anatomically remote and context-sensitive human brain lesions. AB - Functional neuroimaging was used to investigate how lesions to the Broca's area impair neuronal responses in remote undamaged cortical regions. Four patients with speech output problems, but relatively preserved comprehension, were scanned while viewing words relative to consonant letter strings. In normal subjects, this results in left lateralized activation in the posterior inferior frontal, middle temporal, and posterior inferior temporal cortices. Each patient activated normally in the middle temporal region but abnormally in the damaged posterior inferior frontal cortex and the undamaged posterior inferior temporal cortex. In the damaged frontal region, activity was insensitive to the presence of words but in the undamaged posterior inferior temporal region, activity decreased in the presence of words rather than increasing as it did in the normal individuals. The reversal of responses in the left posterior inferior temporal region illustrate the context-sensitive nature of the abnormality and that failure to activate the left posterior temporal region could not simply be accounted for by insufficient demands on the underlying function. We propose that, in normal individuals, visual word presentation changes the effective connectivity among reading areas and, in patients, posterior temporal responses are abnormal when they depend upon inputs from the damaged inferior frontal cortex. Our results serve to introduce the concept of dynamic diaschisis; the anatomically remote and context-sensitive effects of focal brain lesions. Dynamic diaschisis reveals abnormalities of functional integration that may have profound implications for neuropsychological inference, functional anatomy and, vicariously, cognitive rehabilitation. PMID- 11388917 TI - Perceptual and semantic components of memory for objects and faces: a pet study. AB - Previous studies have suggested differences in the neural substrates of recognition memory when the contributions of perceptual and semantic information are manipulated. In a within-subjects design PET study, we investigated the neural correlates of the following factors: material type (objects or faces), semantic knowledge (familiar or unfamiliar items), and perceptual similarity at study and test (identical or different pictures). There was consistent material specific lateralization in frontal and temporal lobe regions when the retrieval of different types of nonverbal stimuli was compared, with objects activating bilateral areas and faces preferentially activating the right hemisphere. Retrieval of memories for nameable, familiar items was associated with increased activation in the left ventrolateral prefrontal cortex, while memory for unfamiliar items involved occipital regions. Recognition memory for different pictures of the same item at study and test produced blood flow increase in left inferior temporal cortex. These results have implications for our understanding of the neural correlates of perceptual and semantic contributions to recognition memory. PMID- 11388918 TI - Inferior temporal neurons show greater sensitivity to nonaccidental than to metric shape differences. AB - It has long been known that macaque inferior temporal (IT) neurons tend to fire more strongly to some shapes than to others, and that different IT neurons can show markedly different shape preferences. Beyond the discovery that these preferences can be elicited by features of moderate complexity, no general principle of (nonface) object recognition had emerged by which this enormous variation in selectivity could be understood. Psychophysical, as well as computational work, suggests that one such principle is the difference between viewpoint-invariant, nonaccidental (NAP) and view-dependent, metric shape properties (MPs). We measured the responses of single IT neurons to objects differing in either a NAP (namely, a change in a geon) or an MP of a single part, shown at two orientations in depth. The cells were more sensitive to changes in NAPs than in MPs, even though the image variation (as assessed by wavelet-like measures) produced by the former were smaller than the latter. The magnitude of the response modulation from the rotation itself was, on average, similar to that produced by the NAP differences, although the image changes from the rotation were much greater than that produced by NAP differences. Multidimensional scaling of the neural responses indicated a NAP/MP dimension, independent of an orientation dimension. The present results thus demonstrate that a significant portion of the neural code of IT cells represents differences in NAPs rather than MPs. This code may enable immediate recognition of novel objects at new views. PMID- 11388919 TI - The time course of visual processing: from early perception to decision-making. AB - Experiments investigating the mechanisms involved in visual processing often fail to separate low-level encoding mechanisms from higher-level behaviorally relevant ones. Using an alternating dual-task event-related potential (ERP) experimental paradigm (animals or vehicles categorization) where targets of one task are intermixed among distractors of the other, we show that visual categorization of a natural scene involves different mechanisms with different time courses: a perceptual, task-independent mechanism, followed by a task-related, category independent process. Although average ERP responses reflect the visual category of the stimulus shortly after visual processing has begun (e.g. 75-80 msec), this difference is not correlated with the subject's behavior until 150 msec poststimulus. PMID- 11388920 TI - Tactile-visual links in exogenous spatial attention under different postures: convergent evidence from psychophysics and ERPs. AB - Tactile-visual links in spatial attention were examined by presenting spatially nonpredictive tactile cues to the left or right hand, shortly prior to visual targets in the left or right hemifield. To examine the spatial coordinates of any crossmodal links, different postures were examined. The hands were either uncrossed, or crossed so that the left hand lay in the right visual field and vice versa. Visual judgments were better on the side where the stimulated hand lay, though this effect was somewhat smaller with longer intervals between cue and target, and with crossed hands. Event-related brain potentials (ERPs) showed a similar pattern. Larger amplitude occipital N1 components were obtained for visual events on the same side as the preceding tactile cue, at ipsilateral electrode sites. Negativities in the Nd2 interval at midline and lateral central sites, and in the Nd1 interval at electrode Pz, were also enhanced for the cued side. As in the psychophysical results, ERP cueing effects during the crossed posture were determined by the side of space in which the stimulated hand lay, not by the anatomical side of the initial hemispheric projection for the tactile cue. These results demonstrate that crossmodal links in spatial attention can influence sensory brain responses as early as the N1, and that these links operate in a spatial frame-of-reference that can remap between the modalities across changes in posture. PMID- 11388921 TI - From knowing what to knowing where: modeling object-based attention with feedback disinhibition of activation. AB - We propose a neural model of visual object-based attention in which the identity of an object is used to select its location in an array of objects. The model is based on neural activity observed in visual search tasks performed by monkeys. In the model, the identity of the object (target) is selected in the higher areas of the ventral stream by means of a cue. Feedback activation from these higher areas carries information about the identity of the target to the (lower) retinotopic areas of the ventral stream. In these areas, the feedback activation interacts with feedforward activation produced by the object array. The interaction occurs in local microcircuits, and results in a selective activation on locations in the retinotopic areas of the visual stream that correspond to the location of the target in the object array. The selective activation consists of a form of gain control, produced by disinhibition. Transmitted to the dorsal stream, this activation directs spatial attention to the location of the target. In this way, an action directed at the target can be generated. PMID- 11388922 TI - Conjoining three auditory features: an event-related brain potential study. AB - The mechanisms of auditory feature processing and conjunction were examined with event-related brain potential (ERP) recording in a task in which participants responded to target tones defined by the combination of location, frequency, and duration features amid distractor tones varying randomly along all feature dimensions. Attention effects were isolated as negative difference (Nd) waves by subtracting ERPs to tones with no target features from ERPs to tones with one, two, or three target features. Nd waves were seen to all tones sharing a single feature with the target, including tones sharing only target duration. Nd waves associated with the analysis of frequency and location features began at latencies of 60 msec, whereas Nd-Duration waves began at 120 msec. Nd waves to tones with single target features continued until 400+ msec, suggesting that once begun, the analysis of tone features continued exhaustively to conclusion. Nd Frequency and Nd-Human Location waves had distinct scalp distributions, consistent with generation in different auditory cortical areas. Three stages of feature processing were identified: (1) Parallel feature processing (60-140 msec): Nd waves combined linearly, such that Nd-wave amplitudes following tones with two or three target features were equal to the sum of the Nd waves elicited by tones with only one target feature. (2) Conjunction-specific (CS) processing (140-220 msec): Nd amplitudes were enhanced following tones with any pair of attended features. (3) Target-specific (TS) processing (220-300 msec): Nd amplitudes were specifically enhanced to target tones with all three features. These results are consistent with a facilitatory interactive feature analysis (FIFA) model in which feature conjunction is associated with the amplified processing of individual stimulus features. Activation of N-methyl-D-aspartate (NMDA) receptors is proposed to underlie the FIFA process. PMID- 11388923 TI - An electrophysiological analysis of the time course of conceptual and syntactic encoding during tacit picture naming. AB - A central question in psycholinguistic research is when various types of information involved in speaking (conceptual/semantic, syntactic, and phonological information) become available during the speech planning process. Competing theories attempt to distinguish between parallel and serial models. Here, we investigated the relative time courses of conceptual and syntactic encoding in a tacit picture-naming task via event-related brain potential (ERP) recordings. Participants viewed pictures and made dual-choice go/no-go decisions based on conceptual features (whether the depicted item was heavier or lighter than 500 g) and syntactic features (whether the picture's German name had feminine or masculine syntactic gender). In support of serial models of speech production, both the lateralized readiness potential, or LRP (related to response preparation), and the N200 (related to response inhibition) measures indicated that conceptual processing began approximately 80 msec earlier than syntactic processing. PMID- 11388924 TI - When in doubt, do it both ways: brain evidence of the simultaneous activation of conflicting motor responses in a spatial stroop task. AB - Response competition is often considered an important contributor to the delayed reaction to stimuli for which physical and semantic information are in conflict ("Stroop" effect). Response competition implies that brain areas associated with correct and incorrect responses (e.g., left and right motor cortices) should be simultaneously activated in conflict conditions. However, there is at present little direct evidence of this phenomenon, in part because of the paucity of brain imaging techniques that can independently monitor the time course of activation of adjacent brain areas, such as the motor areas. In the present study, we show that the event-related optical signal (EROS) can provide these types of data. The results confirm the prediction that conflict trials elicit simultaneous activation of both motor cortices, whereas nonconflict trials elicit brain activity only in the contralateral motor cortex. These data support a parallel view of the human information processing system. PMID- 11388925 TI - Identifying rate-limiting nodes in large-scale cortical networks for visuospatial processing: an illustration using fMRI. AB - With the advent of functional neuroimaging techniques, in particular functional magnetic resonance imaging (fMRI), we have gained greater insight into the neural correlates of visuospatial function. However, it may not always be easy to identify the cerebral regions most specifically associated with performance on a given task. One approach is to examine the quantitative relationships between regional activation and behavioral performance measures. In the present study, we investigated the functional neuroanatomy of two different visuospatial processing tasks, judgement of line orientation and mental rotation. Twenty-four normal participants were scanned with fMRI using blocked periodic designs for experimental task presentation. Accuracy and reaction time (RT) to each trial of both activation and baseline conditions in each experiment was recorded. Both experiments activated dorsal and ventral visual cortical areas as well as dorsolateral prefrontal cortex. More regionally specific associations with task performance were identified by estimating the association between (sinusoidal) power of functional response and mean RT to the activation condition; a permutation test based on spatial statistics was used for inference. There was significant behavioral-physiological association in right ventral extrastriate cortex for the line orientation task and in bilateral (predominantly right) superior parietal lobule for the mental rotation task. Comparable associations were not found between power of response and RT to the baseline conditions of the tasks. These data suggest that one region in a neurocognitive network may be most strongly associated with behavioral performance and this may be regarded as the computationally least efficient or rate-limiting node of the network. PMID- 11388926 TI - Diagnosis and Management of Orthopedic Problems in Children with Cerebral Palsy. AB - Cerebral palsy is a term applied to a variety of clinical syndromes that have as a common feature the abnormal control of motor function by the brain, resulting in disorders of movement and posturing. Sensory function is often affected as well. Cerebral palsy afflicts the immature brain (under 2 years of age) and usually produces neuropathologic changes that do not progress with increasing age. Because myelination of the brain is a progressive process that begins late in the third trimester and continues into adolescence in a well defined pattern, only after the neuronal pathways from brain lesions have become myelinated and the pattern of abnormality determined, can the brain lesions forming the basis of cerebral palsy be detected. PMID- 11388927 TI - Imaging Considerations in Pediatric Skeletal Malignancy. AB - Musculoskeletal neoplasms are rare entities in the pediatric population. Imaging, however, is essential in assisting with the pathologic diagnosis, in therapeutic planning, in monitoring the response to therapy, and in assessing complications of therapy. Additionally, imaging is used in the long-term follow-up of the cancer patient following therapy as surveillance for recurrence. Special considerations in imaging the pediatric patient include issues of child development and behavior which may require modification of routine adult protocols as well as different disease entities that are seen routinely in the adult population. PMID- 11388928 TI - Child Abuse: Imaging Findings Pertaining to the Musculoskeletal System. AB - Both underrecognized and underreported, child abuse has a spectrum of findings on imaging of the musculoskeletal structures that may permit the radiologist to first suggest the occurrence of child abuse or to help support clinical concern for a diagnosis of child abuse. This article will discuss the approach to imaging of the musculoskeletal system in suspected child abuse, illustrate many of the more common imaging findings, and present imaging pitfalls and alternative diagnostic considerations. Injuries to the brain, spinal cord, gastrointestinal and genitourinary systems, and cardiopulmonary structures are outside the scope of this article. PMID- 11388929 TI - Developmental Dysplasia of the Pediatric Hip with Emphasis on Sonographic Evaluation. AB - Developmental dislocation and/or dysplasia of the hip (DDH), formerly referred to as congenital dislocation of the hip, is a frequent problem of the neonatal and infant hip. At that age, the femoral head and acetabulum consist of cartilage components that are not visible on plain radiography but readily identified by ultrasound. Realtime sonography allows for assessment of the hip in multiple planes, both at rest and with movement. Ultrasound can replace radiographic studies and thereby reduce radiation exposure to the young infant. PMID- 11388930 TI - Pediatric Bone Densitometry: Technical Issues. AB - Equipment and software used for dual energy x-ray absorptiometry (DXA) was developed for adults. As use of the method grows in pediatrics, understanding technical issues is important for those performing and interpreting DXA studies in infants, children, and adolescents. Detection of very low bone mineral density requires alteration of automated scan techniques. Normal values based on chronologic age have been published. The Z score becomes the more meaningful statistic as opposed to the T score used in reporting adult DXA measurement of bone mineral density. PMID- 11388931 TI - Legg-Calve-Perthes Disease: Imaging Evaluation and Management. AB - Legg-Calve-Perthes disease reflects avascular necrosis of the proximal femoral epiphysis and growth plate in children age 4 to 8 years typically. The most likely etiology is vascular deficiency to the epiphysis and growth plate. The pathologic stages consist of necrosis, resorption, reossification, and remodeling. Radiologic findings reflect the pathologic stages. Containment of the femoral head with the acetabulum is the most important component of treatment, with preservation of range of motion also indicated in most patients. Surgical options for treatment include varus or valgus femoral osteotomy, innominate osteotomy, and shelf arthroplasty. PMID- 11388932 TI - Prediction of hyperkalemia in dogs from electrocardiographic parameters using an artificial neural network. AB - OBJECTIVE: To predict severe hyperkalemia from single electrocardiogram (ECG) tracings. METHODS: Ten conditioned dogs each underwent this protocol three times: Under isoflurane anesthesia, 2 mEq/kg/hr of potassium chloride was given intravenously until P-waves were absent from the ECG and ventricular rates decreased > or =20% in < or =5 minutes. Serum potassium levels (K(+)) were measured at regular intervals with concurrent digital storage of lead II of the surface ECG. A three-layer artificial neural network with four hidden nodes was trained to predict K(+) from 15 separate elements of corresponding ECG data. Data were divided into a training set and a test set. Sensitivity, specificity, and diagnostic accuracy for recognizing hyperkalemia were calculated for the test set based on a prospectively defined K(+) = 7.5. RESULTS: The model produced data for 189 events; 139 were placed in the training set and 50 in the test set. The test set had 37 potassium levels at or above 7.5 mmol/L. The neural network had a sensitivity of 89% (95% CI = 75% to 97%) and a specificity of 77% (95% CI = 46% to 95%) in recognizing these. The positive likelihood ratio was 3.87. Overall accuracy of this model was 86% (95% CI = 73% to 94%). Mean (+/-SD) difference between predicted and actual K(+) values was 0.4 +/- 2.0 (95% CI = -0.2 to 1.0). CONCLUSIONS: An artificial neural network can accurately diagnose experimental hyperkalemia using ECG parameters. Further work could potentially demonstrate its usefulness in bedside diagnosis of human subjects. PMID- 11388933 TI - Do football helmets reduce acceleration of impact in blunt head injuries? AB - Several recent studies suggest that acceleration of the head at impact during sporting activities may have a detrimental effect on cognitive function. Reducing acceleration of impact in these sports could reduce neurologic sequelae. OBJECTIVE: To measure the effectiveness of a regulation football helmet to reduce acceleration of impact for both low- and moderate-force impacts. METHODS: An experimental paired study design was used. Male volunteers between 16 and 30 years of age headed soccer balls traveling approximately 35 miles per hour bareheaded and with a helmet. An intraoral accelerometer worn inside a plastic mouthpiece measured acceleration of the head. The helmet also had an accelerometer placed inside the padding. For more forceful impacts, cadaver heads, both with and without helmets, were instrumented with intraoral (IO) and intracranial (IC) accelerometers and struck with a pendulum device. Simultaneous IO and IC accelerations were measured and compared between helmeted and unhelmeted cadaver heads. The main outcome was mean peak acceleration of the head and/or brain associated with low- and moderate-force impacts with and without protective headgear. RESULTS: Mean peak Gs, measured by the mouthpiece accelerometer, were significantly reduced when the participants heading soccer balls were wearing a helmet (7.7 Gs with vs 19.2 Gs without, p = 0.01). Wearing a helmet also significantly lowered the peak Gs measured intraorally and intracranially in cadavers subjected to moderate-force pendulum impacts: 28.7 Gs with vs 62.6 Gs without, p < 0.001; and 56.4 Gs with vs 81.6 Gs without, p < 0.001, respectively. CONCLUSIONS: A regulation football helmet substantially reduced the peak Gs associated with "heading" a soccer ball traveling at moderately high velocities. A helmet was also effective in reducing the peak acceleration both intraorally and intracranially for impacts significantly more forceful than heading a soccer ball. PMID- 11388934 TI - Can cardiac sonography and capnography be used independently and in combination to predict resuscitation outcomes? AB - OBJECTIVE: To measure the ability of cardiac sonography and capnography to predict survival of cardiac arrest patients in the emergency department (ED). METHODS: Nonconsecutive cardiac arrest patients prospectively underwent either cardiac ultrasonography alone or in conjunction with capnography during cardiopulmonary resuscitation at two community hospital EDs with emergency medicine residency programs. Cardiac ultrasonography was carried out using the subxiphoid view during pauses for central pulse evaluation and end-tidal carbon dioxide (ETCO(2)) levels were monitored by a mainstream capnograph. A post resuscitation data collection form was completed by each of the participating clinicians in order to assess their impressions of the facility of performance and benefit of cardiac sonography during nontraumatic cardiac resuscitation. RESULTS: One hundred two patients were enrolled over a 12-month period. All patients underwent cardiac sonographic evaluation, ranging from one to five scans, during the cardiac resuscitation. Fifty-three patients also had capnography measurements recorded. The presence of sonographically identified cardiac activity at any point during the resuscitation was associated with survival to hospital admission, 11/41 or 27%, in contrast to those without cardiac activity, 2/61 or 3% (p < 0.001). Higher median ETCO(2) levels, 35 torr, were associated with improved chances of survival than the median ETCO(2) levels for nonsurvivors, 13.7 torr (p < 0.01). The multivariate logistic regression model, which evaluated the combination of cardiac ultrasonography and capnography, was able to correctly classify 92.4% of the subjects; however, of the two diagnostic tests, only capnography was a significant predictor of survival. The stepwise logistic regression model, summarized by the area under the receiver operator curve of 0.9, furthermore demonstrated that capnography is an outstanding predictor of survival. CONCLUSIONS: Both the sonographic detection of cardiac activity and ETCO(2) levels higher than 16 torr were significantly associated with survival from ED resuscitation; however, logistic regression analysis demonstrated that prediction of survival using capnography was not enhanced by the addition of cardiac sonography. PMID- 11388936 TI - Outcome in cardiac arrest patients found to have cardiac standstill on the bedside emergency department echocardiogram. AB - Patients presenting in cardiac arrest frequently have poor outcomes despite heroic resuscitative measures in the field. Many emergency medical systems have protocols in place to stop resuscitative measures in the field; however, further predictors need to be developed for cardiac arrest patients brought to the emergency department (ED). OBJECTIVE: To examine the predictive value of cardiac standstill visualized on bedside ED echocardiograms during the initial presentations of patients receiving cardiopulmonary resuscitation (CPR). METHODS: The study took place in a large urban community hospital with an emergency medicine residency program and a high volume of cardiac arrest patients. As part of routine care, all patients arriving with CPR in progress were subject to immediate and brief subxiphoid or parasternal cardiac ultrasound examination. This was followed by brief repeat ultrasound examination during the resuscitation when pulses were checked. A 2.5-MHz phased-array probe was used for imaging. Investigators filled out standardized data sheets. Examinations were taped for review. Statistical analysis included descriptive statistics, positive and negative predictive values, and likelihood ratios. RESULTS: One hundred sixty nine patients were enrolled in the study. One hundred thirty-six patients had cardiac standstill on the initial echocardiogram. Of these, 71 patients had an identifiable rhythm on monitor. No patient with sonographically identified cardiac standstill survived to leave the ED regardless of his or her initial electrical rhythm. Cardiac standstill on echocardiogram resulted in a positive predictive value of 100% for death in the ED, with a negative predictive value of 58%. CONCLUSIONS: Patients presenting with cardiac standstill on bedside echocardiogram do not survive to leave the ED regardless of their electrical rhythms. This finding was uniform regardless of downtime. Although larger studies are needed, this may be an additional marker for cessation of resuscitative efforts. PMID- 11388937 TI - Emergency department management of patients with seizures: a multicenter study. AB - OBJECTIVES: Patients with seizure disorders are common in the emergency department (ED), yet little is known regarding the management of these patients. This study was performed to define the frequency of patients with seizure disorders in the ED patient population and to determine possible seizure etiologies, characteristics of diagnostic activities, treatments, and dispositions. METHODS: Twelve EDs monitored all patients with a chief complaint related to seizure disorders presenting over 18.25 days (5% of the calendar year) in late 1997. Retrospective chart review was used to gather specific data regarding these consecutive cases. RESULTS: Of the 31,508 patients who presented to these 12 EDs during the study period, 368 (1.2%) had complaints related to seizures. Three hundred sixty-two charts were available for analysis and make up the study population. Two hundred fifty-seven (71%) utilized emergency medical services (EMS) for transport and care. Advanced care, including intravenous access, laboratory work, cardiac monitoring, or oxygen administration, was utilized in 304 (84%) patients. Antiepileptic drugs were given in 199 (55%) patients. Ethanol withdrawal or low antiepileptic drug levels were implicated as contributing factors in 177 (49%) of patients. New-onset seizures were thought to be present in 94 (26%) patients. Status epilepticus occurred in only 21 (6%) patients. Ninety-eight (27%) of all patients were admitted to the hospital. CONCLUSIONS: Patients with presenting complaints related to seizures are frequent in the ED population and make considerable demands on EMS and ED resources. Six percent of patients with seizure-related presentations were in status epilepticus and more than a fourth of all patients required hospitalization. PMID- 11388938 TI - Sledding injuries in patients presenting to the emergency department in a northern city. AB - OBJECTIVES: Sledding is a common recreational activity in northern communities. The objective of this study was to examine the frequency and nature of sledding injuries (SIs) in patients presenting to emergency departments (EDs). METHODS: The data were derived from a cohort of patients treated at all five EDs in an urban Canadian health region over a two-year period. Following chart review, consenting patients were interviewed by telephone about their sledding activities and the circumstances surrounding the injury. RESULTS: Three hundred twenty-eight patients were correctly coded as having SIs, with 212 patients (65%) reached during the follow-up survey. The median age of those with SIs was 12 years (IQR = 8, 21), and 206 (59%) were male. Injury rates peaked in the 10--14-year age group (87/100,000) for boys and in the 5--9-year age group (75/100,000) for girls. Most patients stated they were drivers (75%), fewer than half were thrown from the sled (42%), and fewer than half (44%) were sledding on community-designated sledding hills at the time of injury. Injuries to the lower extremity (32%), upper extremity (31%), and head (13%) were most common. Thirty-seven (11%) patients with SIs were admitted to hospital vs 4% of patients with other sports/recreation injuries (p < 0.05). CONCLUSIONS: Sledding injuries are common and potentially serious wintertime injuries in northern communities, involving primarily younger patients, with a large pre-adolescent group. However, older sledders (>20 years) have poorer outcomes (hospitalization, lost time from work/school) than their younger counterparts. The SIs treated in the ED appear to lead to hospitalization more frequently than other types of sport/recreation injury, and injury prevention strategies appear warranted. PMID- 11388939 TI - Variation in institutional review board responses to a standard protocol for a multicenter clinical trial. AB - Multicenter clinical trials require approval by multiple local institutional review boards (IRBs). The Multicenter Airway Research Collaboration mailed a clinical trial protocol to its U.S. investigators and 44 IRBs ultimately reviewed it. OBJECTIVE: To describe IRB responses to one standard protocol and thereby gain insight into the advantages and disadvantages of local IRB review. METHODS: Two surveys were mailed to participants, with telephone follow-up of nonrespondents. Survey 1 was mailed to 82 investigators across North AMERICA: Survey 2 was mailed to investigators from 44 medical centers in 17 U.S. states. Survey 1 asked about each investigator's local IRB (e.g., frequency of meetings, membership), whereas survey 2 asked about IRB queries and concerns related to the submitted clinical trial. RESULTS: Both surveys had 100% response rate. Investigators submitted applications a median of 58 days (interquartile range [IQR], 40--83) after receipt of the protocol, and IRB approval took an additional 38 days (IQR, 26--62). Although eight applications were approved with little or no changes, IRBs requested an average of 3.5 changes per site. Changes involved study logistics and supervision for 45%, the research process for 43%, and the consent form for 91%. Despite these numerous requests, all eventually approved the basic protocol, including inclusion criteria, intervention, and data collection. CONCLUSIONS: The IRBs showed extreme variability in their initial responses to a standard protocol, but ultimately all gave approval. Almost all IRBs changed the consent form. A national, multicenter IRB process might streamline ethical review and warrants further consideration. PMID- 11388940 TI - Effect of changes in graduate medical education funding on emergency medicine residency programs. AB - OBJECTIVE: To determine whether changes in graduate medical education (GME) funding have had an impact on emergency medicine (EM) residency training programs. METHODS: A 34-question survey was mailed to the program directors (PDs) of all 115 Accreditation Council for Graduate Medical Education (ACGME) accredited EM residency programs in the United States in the fall of 1998, requesting information concerning the impact of changes in GME funding on various aspects of the EM training. The results were then compared with a similar unpublished survey conducted in the fall of 1996. RESULTS: One hundred one completed surveys were returned (88% response rate). Seventy-one (70%) of the responding EM residency programs were PGY-I through PGY-III, compared with 55 (61%) of the responding programs in 1996. The number of PGY-II through PGY-IV programs decreased from 25 (28%) of responding programs in 1996 to 17 (16%). The number of PGY-I through PGY-IV programs increased slightly (13 vs 10); the number of EM residency positions remained relatively stable. Fifteen programs projected an increase in their number of training positions in the next two years, while only three predicted a decrease. Of the respondents, 56 programs reported reductions in non-EM residency positions and 35 programs reported elimination of fellowship positions at their institutions. Only four of these were EM fellowships. Forty-six respondents reported a reduction in the number of non-EM residents rotating through their EDs, and of these, 11 programs reported this had a moderate to significant effect on their ability to adequately staff the ED with resident physicians. Sixteen programs limited resident recruitment to only those eligible for the full three years of GME funding. Eighty-seven EM programs reported no change in faculty size due to funding issues. Sixty-two programs reported no change in the total number of hours of faculty coverage in the ED, while 34 programs reported an increase. Three EM programs reported recommendations being made to close their residency programs in the near future. CONCLUSIONS: Changes in GME funding have not caused a decrease in the number of existing EM residency and fellowship training positions, but may have had an impact in other areas, including: an increase in the number of EM programs structured in a PGY-I through PGY-III format (with a corresponding decrease in the number of PGY-II through PGY-IV programs); a decrease in the number of non-EM residents rotating through the ED; restriction of resident applicants who are ineligible for full GME funding from consideration by some EM training programs; and an increase in the total number of faculty clinical hours without an increase in faculty size. PMID- 11388941 TI - Emergency medicine standardized letter of recommendation: predictors of guaranteed match. AB - OBJECTIVE: The Council of Emergency Medicine Residency Directors (CORD) standardized letter of recommendation (SLOR) has become a common, reliable, and useful tool in the evaluation of emergency medicine (EM) applicants. A "guaranteed match" (GM) is the SLOR's bottom-line superlative response. It is also the SLOR's least common superlative response. Because candidates receiving a GM are a select group, the authors thought it would be useful to identify SLOR information that predicts a GM recommendation. METHODS: This was a secondary analysis of a database of all EM SLORs submitted to a single EM residency during the 1998--1999 application cycle to one EM residency program. Response to GM and 16 data points in the background/qualification sections were analyzed by chi square, univariate analysis, and logistic regression. RESULTS: Four hundred eleven SLORs were analyzed. Qualification information was more predictive than background information for applicants receiving a GM. The highest univariate odds ratios for background information were "staff author" (OR = 1.7, 1.0--2.8), "extended contact" (OR = 2.2, 1.0--4.5), "clinical contact outside the ED" (OR = 3.0, 1.5--5.9), and "honors on EM rotation" (OR = 5.4, 3.0--9.8). The highest univariate odds ratios for qualification information were "outstanding differential diagnosis ability" (OR = 10.1, 5.8--17.4), "outstanding work ethic" (OR = 13.1, 5.2--33.3), and "outstanding global assessment" (OR = 58, 24.2--139). Logistic regression analysis demonstrated "outstanding global assessment" (p < 0.000; r = 0.92) and "outstanding work ethic" (p = 0.028; r = 0.71) to be statistically predictive of GM. CONCLUSIONS: There were both background and qualification data points predictive of a "guaranteed match." Qualification information had a greater predictive value than background information. Medical student applicants, letter writers, and letter evaluators may find this information useful when dealing with SLORS. PMID- 11388942 TI - Searching for a predictive rule for terminating cardiopulmonary resuscitation. PMID- 11388943 TI - Why a new model? PMID- 11388945 TI - Monthly, weekly, and daily patterns in the incidence of congestive heart failure. AB - OBJECTIVE: To determine whether there are patterns in the incidence of emergency department (ED) visits for congestive heart failure (CHF) by month of the year, day of the week, or hour of the day. METHODS: This was a retrospective analysis of a computerized billing database of ED visits, involving seven northern New Jersey hospitals EDS: Consecutive patients seen by emergency physicians over an 11-year period (January 1, 1988--December 31, 1998) were included. Chi-square tests were used to evaluate for significant differences (p < 0.05). RESULTS: There were a total of 2,370,233 patients in the database, of whom 26,224 had a primary ED diagnosis of CHF. The chi-square test rejected uniformity for month of the year, for day of the week, and for hour of the day (p < 0.0001). Visits for CHF were increased in the winter months. Compared with the average of the other months, December was the highest (14.3% above, p < 0.0001) and August was the lowest (15.5% below, p < 0.0001). There was also a day-of-the-week variation. Compared with the average of the other days, Monday was the highest (14.5% above, p < 0.0001) and Saturday was the lowest (9.6% below, p < 0.0001). There was also an hour-of-the-day pattern, with a rapid rise after 8 AM and a downtrend after 3 PM. CONCLUSIONS: These data revealed a higher incidence of ED visits for CHF in the winter months, on Mondays, and during the hours of 8 AM to 3 PM. In comparison with previous studies, these data revealed a similar pattern by month of the year and a different pattern by hour of the day. PMID- 11388946 TI - The November special issue on errors. PMID- 11388948 TI - The search for the wandering thymostat: a review of some developments in bipolar disorder research. PMID- 11388947 TI - Rattlesnake bites. PMID- 11388949 TI - Bipolar disorder: leads from the molecular and cellular mechanisms of action of mood stabilisers. AB - Background New research is dramatically altering our understanding of the molecular mechanisms underlying neuronal communication. Aim To elucidate the molecular mechanisms underlying the therapeutic effects of mood stabilisers. Method Results from integrated clinical and laboratory studies are reviewed. Results Chronic administration of lithium and valproate produced a striking reduction in protein kinase C (PKC) isozymes in rat frontal cortex and hippocampus. In a small study, tamoxifen (also a PKC inhibitor) had marked antimanic efficacy. Both lithium and valproate regulate the DNA binding activity of the activator protein 1 family of transcription factors. Using mRNA differential display, it was also shown that chronic administration of lithium and valproate modulates expression of several genes. An exciting finding is that of a robust elevation in the levels of the cytoprotective protein, bcl-2. Conclusions The results suggest that regulation of signalling pathways may play a major part in the long-term actions of mood stabilisers. Additionally, mood stabilisers may exert underappreciated neuroprotective effects. PMID- 11388950 TI - Neuropsychology of bipolar disorder. AB - Background Although the presence of wide-ranging neuropsychological deficits in individuals with major depression is well established, few studies have investigated the nature of cognitive impairment in patients with bipolar disorder. Aims To review research of the neuropsychology of bipolar disorder, with special attention to the relationship between mood and cognitive functioning. Method Literature review. Results Findings generally demonstrate mania-related impairments on conventional neuropsychological tests, with direct comparisons of patients with mania and patients with depression failing to find group differences. More recent work has sought to differentiate these disorders by employing tasks with affective components. This research has demonstrated biases for processing positive and negative stimuli in patients with mania and depression, respectively. Conclusions Future studies, employing tasks that require cognitive and emotional processing, should improve our understanding of the deficits observed in depression and mania. Neuroimaging studies of the neural regions that underlie cognitive processing of affective meaning suggest that the medial and orbitofrontal prefrontal cortex may be particularly involved. PMID- 11388951 TI - Molecular genetics of bipolar disorder. AB - Background A robust body of evidence from family, twin and adoption studies demonstrates the importance of genes in the pathogenesis of bipolar disorder. Recent advances in molecular genetics have made it possible to identify these susceptibility genes. Aims To present an overview for clinical psychiatrists. Method Review of current molecular genetics approaches and emerging findings. Results Occasional families may exist in which a single gene plays a major role in determining susceptibility, but the majority of bipolar disorder involves more complex genetic mechanisms such as the interaction of multiple genes and environmental factors. Molecular genetic positional and candidate gene approaches are being used for the genetic dissection of bipolar disorder. No gene has yet been identified but promising findings are emerging. Regions of interest include chromosomes 4p16, 12q23-q24, 16p13, 21q22, and Xq24-q26. Candidate gene association studies are in progress but no robust positive findings have yet emerged. Conclusion It is almost certain that over the next few years the identification of bipolar susceptiblity genes will have a major impact on our understanding of disease pathophysiology. This is likely to lead to major improvements and treatment in patient care, but will also raise important ethical issues. PMID- 11388952 TI - Genetic studies of bipolar affective disorder in large families. AB - Background Genetic factors are known to be important in the aetiology of bipolar disorder. Aims To review linkage studies in extended families multiply affected with bipolar disorder. Method Selective review of linkage studies of bipolar disorder emphasising the gains and drawbacks of studying large multiply-affected families and comparing the statistical methods used for data analysis. Results Linkage of bipolar disorder to several chromosome regions including 4p, 4q, 10p, 12q, 16p, 18q, 21q and Xq has first been reported in extended families. In other families chromosomal rearrangements associated with affective illnesses provide signposts to the location of disease-related genes. Statistical analyses using variance component methods can be applied to extended families, require no prior knowledge of the disease inheritance, and can test multilocus models. Conclusion Studying single large pedigrees combined with variance component analysis is an efficient and effective strategy likely to lead to further insights into the genetic basis of bipolar disorders. PMID- 11388954 TI - Neuroanatomical studies on bipolar disorder. AB - Background Neuroimaging data showing structural and functional brain abnormalities in mood disorders suggest that brain alterations at the neurohistological level may underlie the macropathology seen by imaging in vivo. Aims To summarise recent post-mortem studies on affective disorders, with a focus on bipolar disorder. Method Literature review and discussion of results from volumetric, cyto-architectural and immunohistochemical analyses. Results Basal ganglia are smaller in patients with depression irrespective of diagnostic polarity. In addition, higher neuron numbers have been reported in the locus caeruleus of patients with bipolar disorder compared with those with unipolar depression. Patients with bipolar as well as unipolar illness show subtle structural deficits in the dorsal raphe. Histological data are consistent with a regional reduction in the synthesis of noradrenalin and serotonin, which appears to be compensated by antidepressants. Conclusion Preliminary results suggest that, aside from functional dysregulation, subtle structural abnormalities in the brain may contribute to the pathogenesis of mood disorders. PMID- 11388955 TI - Use of antipsychotic drugs and lithium in mania. AB - Background Studies highlighting the difficulties associated with lithium suggest that the role of antipsychotic drugs and mood stabilisers in bipolar disorder should be reconsidered. Aims To review the efficacy and mode of action of antipsychotic drugs in mania, and to consider the differences between official guidelines and routine clinical practice in the use of these agents for mania. Method Review of research, guideline-and practice-based literature. Results Guidelines recommend lithium or valproate as first-line treatments for mania, and antipsychotic agents only as 'adjuncts' for agitation, dangerous behaviour or psychosis. However, in routine practice, antipsychotic drugs are often prescribed. The effectiveness of these agents in mania has been established by several studies; newer atypical compounds demonstrate antimanic efficacy with a reduced incidence of neurological side-effects. Conclusion Antipsychotic drugs are important in the treatment of bipolar disorder and mania. Future studies should evaluate the long-term efficacy and safety of newer atypical antipsychotic agents, and the place of anticonvulsants in combination with antipsychotics in bipolar disorder. PMID- 11388956 TI - Evolving methodologies in bipolar disorder maintenance research. AB - Background During the development of a new treatment for bipolar disorder, maintenance studies are used to evaluate the ability of the putative mood stabiliser to prevent relapse and recurrence of further episodes. Comparisons with the early bipolar disorder maintenance studies indicate that the methodologies of recent trials have evolved substantially. Aims To review the methods used in the first- and second-generation maintenance studies, highlighting the differences of the various designs. Method Literature review. Results Methods that have evolved the most include patient enrolment, randomisation schemes and the use of outcome measures and statistical analyses. In addition, regulatory and commercial issues have also influenced study design. Conclusion There is little consensus on the methodology of bipolar disorder maintenance studies. As the integration of newer therapies into routine clinical practice is dependent on the evidence from controlled studies, it is essential that future maintenance trials in bipolar disorder achieve adequate methodological rigour without sacrificing overall feasibility. PMID- 11388957 TI - Cognitive therapy as an adjunct to medication in bipolar disorder. AB - Background There is increasing support for the use of cognitive behaviour therapy as an adjunct to medication for patients with bipolar disorder. Aims To explore current psychological models of bipolar disorder, describing the clinical rationale for using cognitive therapy and providing a brief overview of the approach. Method Results from outcome studies are discussed. Results Preliminary findings indicate that cognitive therapy may be beneficial for patients with bipolar disorder. The collaborative, educational style of cognitive therapy, the use of a stepwise approach and of guided discovery is particularly suitable for patients who wish to take an equal and active role in their therapy. Conclusions Randomised, controlled trials of cognitive therapy in bipolar disorder are required to establish the short-term and long-term benefits of therapy, and whether any reported health gain exceeds that of treatment as usual. PMID- 11388960 TI - Long-term clinical effectiveness of lithium maintenance treatment in types I and II bipolar disorders. AB - Background The effectiveness of lithium is being questioned increasingly and requires clarification. Aims To assess the effectiveness of lithium treatment in depression and mania, syndromal types I and II, with predominantly mixed or psychotic episodes or rapid cycling, during treatment resumed following discontinuation, and across three decades. Method The longitudinal course of 360 patients with bipolar disorder compliant with lithium treatment for at least I year and without comorbidity for substance use disorder was reviewed. Results Risk of single-episode recurrences, a common index of treatment failure, was similar to that in other reports. Both episode frequency and 'time ill' improved more in type II than type I cases. Reduced morbidity during treatment was similar in patients with mixed or psychotic episodes, or rapid cycling, and in less complex cases. Retreatment yielded minor decrements in response, and there was no tendency for lesser responses in more recent years. Conclusions Based on overall affective morbidity, long-term lithium treatment in compliant patients without comorbid substance use disorder, though imperfect, remains effective, even in subgroups of supposedly poor prognosis. PMID- 11388961 TI - Bipolar disorder: clinical uncertainty, evidence-based medicine and large-scale randomised trials. AB - Background The increasing use of the methods of evidence-based medicine to keep up-to-date with the research literature highlights the absence of high-quality evidence in many areas in psychiatry. Aims To outline current uncertainties in the maintenance treatment of bipolar disorder and to describe some of the decisions involved in designing a large simple trial. Method We describe some of the strategies of evidence-based medicine, and how they can be applied in practice, focusing specifically on the area of bipolar disorder. Results One of the key clinical uncertainties in the treatment of bipolar disorder is the place of maintenance drug treatments and their relative efficacy. A large-scale study, the Bipolar Affective DISORDER: Lithium Anticonvulsant Evaluation (BALANCE) trial, is proposed to compare the effectiveness of lithium, valproate and the combination of lithium and valproate. Conclusions Providing reliable answers to key clinical questions in psychiatry will require new approaches to clinical trials. These will need to be far larger than previously appreciated and will therefore need to be collaborative ventures involving front-line clinicians. PMID- 11388962 TI - The distinction between mental and physical illness. PMID- 11388963 TI - Role of psychiatrists in the prediction and prevention of suicide: a perspective from north-east Scotland. PMID- 11388964 TI - Community mental health team management in severe mental illness: a systematic review. AB - BACKGROUND: Community mental health teams are now generally recommended for the management of severe mental illness but a comparative evaluation of their effectiveness is lacking. AIMS: To assess the benefits of community mental health team management in severe mental illness. METHOD: A systematic review was conducted of community mental health team management compared with other standard approaches. RESULTS: Community mental health team management is associated with fewer deaths by suicide and in suspicious circumstances (odds ratio=0.32, 95% Cl 0.09-1.12), less dissatisfaction with care (odds ratio=0.34, 95% Cl 0.2-0.59) and fewer drop-outs (odds ratio=0.61, 95% Cl 0.45-0.83). Duration of in-patient psychiatric treatment is shorter with community team management and costs of care are less, but there are no gains in clinical symptomatology or social functioning. CONCLUSIONS: Community mental health team management is superior to standard care in promoting greater acceptance of treatment, and may also reduce hospital admission and avoid deaths by suicide. This model of care is effective and deserves encouragement. PMID- 11388966 TI - Recovery from psychotic illness: a 15- and 25-year international follow-up study. AB - BACKGROUND: Poorly defined cohorts and weak study designs have hampered cross cultural comparisons of course and outcome in schizophrenia. AIMS: To describe long-term outcome in 18 diverse treated incidence and prevalence cohorts. To compare mortality, 15- and 25-year illness trajectory and the predictive strength of selected baseline and short-term course variables. METHODS: Historic prospective study. Standardised assessments of course and outcome. RESULTS: About 75% traced. About 50% of surviving cases had favourable outcomes, but there was marked heterogeneity across geographic centres. In regression models, early (2 year) course patterns were the strongest predictor of 15-year outcome, but recovery varied by location; 16% of early unremitting cases achieved late-phase recovery. CONCLUSIONS: A significant proportion of treated incident cases of schizophrenia achieve favourable long-term outcome. Sociocultural conditions appear to modify long-term course. Early intervention programmes focused on social as well as pharmacological treatments may realise longer-term gains. PMID- 11388967 TI - Patterns and predictors of hospitalisation in first-episode psychosis. Prospective cohort study. AB - BACKGROUND: Little is known about predictors of hospitalisation in patients with first-episode psychosis. AIMS: To identify the pattern and predictors of hospitalisation of patients with a first psychotic episode making their first contact with specialist services. METHOD: Three-year follow-up of a cohort of 166 patients with a first episode of psychosis making contact with psychiatric services in Nottingham between June 1992 and May 1994. RESULTS: Eighty-eight (53.0%) patients were admitted within 1 week of presentation; 32 (19.3%) were never admitted during the 3 years of follow-up. Manic symptoms at presentation were associated with an increased risk of rapid admission and an increased overall risk of admission; negative symptoms and a longer duration of untreated illness had an increased risk of late admission. CONCLUSIONS: Community-oriented psychiatric services might only delay, rather than prevent, admission of patients with predominantly negative symptoms and a longer duration of untreated illness. First-episode studies based upon first admissions are likely to be subject to selection biases, which may limit their representativeness. PMID- 11388968 TI - Neurodevelopmental indices and the development of psychotic symptoms in subjects at high risk of schizophrenia. AB - BACKGROUND: Neurological 'soft signs' and minor physical anomalies (MPAs) are reported to be more frequent in patients with schizophrenia than in controls. AIMS: To determine whether these disturbances are genetically mediated, and whether they are central to the genesis of symptoms or epiphenomena. METHOD: We obtained ratings in 152 individuals who were antipsychotic drug-free and at high risk, some of whom had experienced psychotic symptoms, as well as 30 first episode patients and 35 healthy subjects. RESULTS: MPAs and Neurological Evaluation Scale (NES) 'sensory integration abnormalities' were more frequent in high-risk subjects than in healthy controls, but there were no reliable differences between high-risk subjects with and without psychotic symptoms. MPAs were most frequent in high-risk subjects with least genetic liability and NES scores showed no genetic associations. CONCLUSIONS: The lack of associations with psychotic symptoms and genetic liability to schizophrenia suggests that soft signs and physical anomalies are nonspecific markers of developmental deviance that are not mediated by the gene(s) for schizophrenia. PMID- 11388969 TI - The Wessex Recent In-Patient Suicide Study, 1. Case-control study of 234 recently discharged psychiatric patient suicides. AB - BACKGROUND: Psychiatric patients have a higher suicide risk following hospital discharge. AIMS: To identify social, clinical and health-care delivery factors in recently discharged patients. METHOD: Retrospective case-control study of 234 patients who died within 1 year of hospital discharge, matched for age, gender, diagnosis and admission period with 431 controls. Odds ratios for identified risk factors were calculated using conditional multiple logistic regression. RESULTS: Independent increased-risk factors were: not being White; living alone; history of deliberate self-harm (DSH); suicidal ideation precipitating admission; hopelessness; admission under different consultant; onset of relationship difficulties; loss of job; in-patient DSH; unplanned discharge; significant care professional leaving/on leave. Reduced-risk factors were: shared accommodation; delusions at admission; misuse of non-prescribed substances; and continuity of contact. CONCLUSIONS: Continuity of contact may reduce suicide risk. Discontinuity of care from a significant professional is associated with increased risk of suicide. PMID- 11388970 TI - The Wessex Recent In-Patient Suicide Study, 2. Case-control study of 59 in patient suicides. AB - BACKGROUND: Psychiatric patients have an elevated risk of suicide while in hospital. AIMS: To compare social, clinical and health-care delivery factors in in-patient and out-patient suicides and their controls. METHOD: Retrospective case-control study of 59 in-patients and 106 controls, matched for age, gender, diagnosis and admission date. Odds ratios were calculated using conditional multiple logistic regression. RESULTS: There were seven independent increased risk factors: history of deliberate self-harm, admission under the Mental Health Act, involvement of the police in admission, depressive symptoms, violence towards property, going absent without leave and a significant care professional being on leave. When compared with out-patient suicides, in-patients were more often female and male in-patients had a psychotic illness. Unlike the out-patient suicides, social factors were not found to be significant. CONCLUSIONS: The characteristics of in-patient and out-patient suicides differ. Identified risk factors have relatively low sensitivity and specificity. PMID- 11388971 TI - Two-year follow-up after a randomised controlled trial of self- and clinician accompanied exposure for phobia/panic disorders. AB - BACKGROUND: Long-term follow-up has rarely been reported after self-exposure therapy for phobias. AIMS: Completion of such a follow-up. METHOD: Two-year follow-up was achieved in 68 (85%) of 80 patients with phobias who had completed a previous 14-week randomised controlled trial comparing therapist-accompanied self-exposure, self-exposure or self-relaxation. Measures were self-reported ratings of symptoms, satisfaction and use of other treatment. RESULTS: Improvement at week 14 was maintained 2 years later. Clinician-accompanied exposure and self-exposure did not differ on any measure. Compliance with self exposure homework during weeks 0-8 predicted more improvement 2 years later. Patients who failed to improve with relaxation by week 14 improved after subsequent crossover to exposure. A need for more treatment for their phobias was still felt by 33 patients (49%). CONCLUSIONS: Patients with phobias maintained their improvement to 2-year follow-up after the end of self-exposure therapy. PMID- 11388972 TI - Sensation-seeking, life events and depression. The Cardiff Depression Study. AB - BACKGROUND: The relationship between adversity and genetic risk factors in depression could be mediated by familial 'hazard prone' traits, as reflected in high levels of sensation-seeking. AIMS: To examine whether high sensation-seeking scores are associated with more adverse life events resulting in depression. METHOD: In a sib-pair design, 108 probands with depression and their siblings and 105 healthy control subjects and their siblings were compared for psychopathology, life events and scores on the Sensation-Seeking Questionnaire (SSQ). RESULTS: The SSQ scores were correlated negatively with depression, were familial and were correlated positively with less severe events, but not the severe events typically associated with depressive onsets. CONCLUSIONS: The SSQ measures a familial personality trait and depression is associated with lower scores. Although high sensation-seeking is associated with a higher rate of life events, these carry little threat. PMID- 11388973 TI - Association of hypotension with positive and negative affect and depressive symptoms in the elderly. AB - BACKGROUND: Previous research associating hypotension with depression has produced inconsistent results. A possible reasons is that depressive symptom scales reflect both high negative affect and low positive affect. AIMS: To examine the association of hypotension with depressive symptoms, negative affect and positive affect. METHOD: Community survey of 340 elderly persons aged 77-99 years. RESULTS: Diastolic hypotension had a weak association with depression, no association with negative affect and a strong inverse association with positive affect. Systolic hypertension was associated with positive affect. Use of antihypertensive medication was independently associated with lower positive affect. CONCLUSIONS: Diastolic hypotension shows a specific association with low positive affect. This association may explain the weak and inconsistent results of earlier studies relating hypotension to depression. PMID- 11388974 TI - Incidence, correlates and predictors of post-traumatic stress disorder in the pregnancy after stillbirth. AB - BACKGROUND: Many women may suffer psychological symptoms after stillbirth and in the subsequent pregnancy. Stillbirth has not been demonstrated previously to be a stressor for post-traumatic stress disorder (PTSD). AIMS: To assess incidence, correlates and predictors of PTSD during and after the pregnancy following stillbirth. METHOD: A cohort study of pregnant women whose previous pregnancy ended in stillbirth. RESULTS: PTSD symptoms were prevalent in the pregnancy following stillbirth. Case-level PTSD was associated with depression, state anxiety and conception occurring closer to loss. Symptoms generally resolved naturally by 1 year post-partum (birth of healthy baby). CONCLUSIONS: Women are vulnerable to PTSD in the pregnancy subsequent to stillbirth, particularly when conception occurs soon after the loss. PMID- 11388975 TI - Value of HoNOS in assessing patient change in NHS psychotherapy and psychological treatment services. AB - BACKGROUND: Little research on the value of Health of the Nation Outcome Scales (HoNOS) has occurred in out-patient settings, particularly psychotherapy services. AIMS: To determine whether HoNOS provides an adequate assessment for psychotherapy services which is sensitive to change. METHODS: HoNOS ratings from 1688 patients from eight out-patient psychotherapy services were collected. Of these, 362 also had ratings post-treatment. Mean scores, pre-to post-treatment differences, and reliable and clinically significant change criteria were calculated for HoNOS items and for total scores. RESULTS: The mean total HoNOS rating was 8.93, which is comparable to psychiatric out-patients. Only three items showed sufficient variability to use in assessing pre- to post-treatment change. CONCLUSIONS: Significant limitations were found in rating items that commonly present to psychotherapists. The lack of variability in most items limits HoNOS's usefulness in this population. PMID- 11388976 TI - Cross-cultural implementation of a Chinese version of the Schedules for Clinical Assessment in Neuropsychiatry (SCAN) in Taiwan. AB - BACKGROUND: There are no published reports of cross-cultural equivalence and interrater reliability at the level of individual symptom items assessed by a semi-structured clinical interview employing operationalised clinician ratings. AIMS: To assess the cross-cultural clinical equivalence and reliability of a Chinese version of the World Health Organization Schedules for Clinical Assessment in Neuropsychiatry (SCAN). METHOD: UK-US and Taiwanese groups of psychiatrists used Chinese and English transcripts of videotape interviews of Taiwanese patients to discuss cross-cultural issues and ratings of SCAN items. Item ratings were compared quantitatively individually and pooled by SCAN section. RESULTS: Chinese equivalents were found for all SCAN items. No between group differences were found for most individual items, but there were differences for some scaled items. Average agreement between the two groups was 69-100%. CONCLUSIONS: Cross-cultural implementation based on SCAN in Taiwan appears valid. PMID- 11388977 TI - Evolutionary psychology revisited. PMID- 11388978 TI - Placebo response in antidepressant trials. PMID- 11388979 TI - Cannabis regimes. PMID- 11388980 TI - Evidence-based psychiatry. PMID- 11388981 TI - Effects of schizophrenia on patients' relatives. PMID- 11388982 TI - Over-representation of Black people in secure psychiatric facilities. PMID- 11388983 TI - Reintroduction of clozapine after diagnosis of lymphoma. PMID- 11388985 TI - The keeping of lunatics in unlicensed houses. PMID- 11388984 TI - Stigma and ineffective legislation. PMID- 11388986 TI - The clinical use of PET--where are we now? PMID- 11388987 TI - Palpable asymmetrical thickening of the breast: a clinical, radiological and pathological study. AB - While management protocols for a discrete palpable breast lump are standardized in most centres, the approach to an area of palpable asymmetrical thickening in the breast has seldom been addressed. A diagnostic algorithm for palpable asymmetrical thickening of the breast was prospectively evaluated in 116 Oriental women, followed by a retrospective review of their mammograms and histology specimens. Most women (86%) were pre-menopausal and 82% complained of a lump. The thickening eventually resolved spontaneously in 93 (80%) women. None of these 93 women developed cancer at a median follow-up of 41 months. A total of 9 (7.8%) cancers were found in the series of 116 women, including two with a lobular component. The occurrence of cancer was more likely when the woman was older than 43 years or when the thickening was marked (p<0.04). Mammographic review showed correlation of the palpable thickening with localized increase in breast tissue density and/or microcalcifications in 18% of cases. Histology review suggested fibrosis as an explanation for the clinical presentation. Although most cases of thickening tend to resolve with time, a significant number of cancers present in this way. A diagnostic approach with early and liberal imaging and biopsy for high risk women is required. PMID- 11388988 TI - T(2) relaxation rate of basal ganglia and cortex in patients with beta thalassaemia major. AB - In thalassaemic patients, neurophysiological disturbances have been associated with high serum ferritin levels and desferrioxamine therapy. In the presence of a magnetic field, ferritin, the main iron storage protein, induces a preferential decrease of the T(2) relaxation time. The purpose of this study was to evaluate thalassaemic patients for brain iron deposition by assessing the T(2) relaxation rate (1/T(2)) of the grey matter. 41 thalassaemic patients (age range 8.5-44 years, mean 24 years) and 58 age- and sex-matched controls were included in the study. Current serum ferritin levels were obtained. The 1/T(2) values of the cortex (motor and temporal) (mean 0.0122 ms(-1), SD 0.0004), putamen (mean 0.0137 ms(-1), SD 0.0004) and caudate nucleus (mean 0.0132 ms(-1), SD 0.0003) were higher in patients compared with the controls (mean 0.0110 ms(-1), SD 0.0004; mean 0.0120 ms(-1), SD 0.0005; mean 0.0117 ms(-1), SD 0.0003, respectively) (p<0.001 for all parameters). No statistically significant differences were found in the globus pallidus. No correlation was found between 1/T(2) and serum ferritin. The higher values of 1/T(2) in the cortex, putamen and caudate nucleus of thalassaemic patients probably reflect a higher iron deposition. The lack of differences in 1/T(2) of the globus pallidus might suggest that even in thalassaemic patients iron cannot exceed a saturation level. PMID- 11388989 TI - Renal allograft vasculopathy: ultrasound findings in a non-human primate model of chronic rejection. AB - The purpose was to determine whether decreased cortical flow detected with power Doppler (PD) ultrasound in renal allografts in cynomolgus monkeys marks the presence or onset of chronic renal allograft vasculopathy. The 2D grey scale and PD ultrasound findings of 24 consecutively implanted non-life-supporting renal allografts in cynomolgus monkeys that underwent either 24 h (n=15) or 48 h (n=9) cold ischaemia times were recorded and compared with the results of histology performed every 2 weeks post-operatively. 13 allografts developed vasculopathies, 10 of which had PD scores equal to 1 (severe reduction of cortical flow). A PD score of 1 occurred in only one instance in the group of allografts without vasculopathies and this was due to necrosis. Allografts without vasculopathies otherwise had either PD scores of 3 (normal flow; n=2) or 2 (reduced flow; n=4). Allografts subjected to 48 h cold ischaemia times were smaller than those with 24 h cold ischaemia times (significant at weeks 5-11, p<0.05), but a reduction in graft size associated with vasculopathies occurred infrequently. In conclusion, the finding of reduced renal cortical flow detected by PD ultrasound during serial examination of non-life-supporting renal allografts is highly supportive of a diagnosis of graft vasculopathy due to arteriolar intimal proliferation, and illustrates an excellent method of monitoring changes in cortical perfusion in allografts in animal models. The combination of findings of reduced or absent cortical flow together with severe graft enlargement is highly suggestive of the presence of not only vasculopathies but also tissue damage and degeneration. PMID- 11388990 TI - Patient and occupational dosimetry in double contrast barium enema examinations. AB - A new and relatively simple method is presented to distribute total dose-area product (DAP) over a number of projections that model exposure during double contrast barium enema (DCBE) examinations. In addition, hitherto unavailable entrance and effective doses to the physician performing the DCBE examination have been determined. DAP, fluoroscopy time, number of images as well as some patient data were collected for 150 DCBE examinations. For a subset of 50 examinations, the distribution of DAP over 12 hypothetical but representative projections was estimated by measuring the entrance dose in the centre of each of these projections during the complete procedure. Effective dose to the patient was obtained using DAP to effective dose conversion coefficients calculated for each of the 12 projections. Exposure of the worker was quantified by measuring the entrance dose at the forehead, neck, arms, right hand and legs. The sex averaged effective dose to the patient per examination was 6.4+/-2.1 mSv (mean+/ SD; n=50) and the corresponding DAP was 44+/-22 Gy cm(2). The effective dose to the worker per examination was 0.52 microGy (n=50), whereas the highest entrance dose of 30+/-25 microGy was found for the right arm. The proposed method for deriving the distribution of total DAP over a set of representative projections is much less time consuming than visual observation of patient exposure, whilst accuracy seems acceptable. Entrance and effective doses per examination for workers in DCBE examinations are very low. For a normal workload, doses remain far below the legally established dose limits. PMID- 11388991 TI - Value of protein-bound radioactive iodine measurements in the management of differentiated thyroid cancer treated with (131)I. AB - Measurement of the protein-bound radioactive iodine level (PBI(131)) in the plasma of patients following (131)I-iodide administration for thyroid cancer has been re-examined in a retrospective study of 171 patient episodes. It is shown that whereas the previously used threshold value for the measurement at 6 days does not correlate well with the 3-day whole body scan, there is good agreement between the scan and the temporal changes in PBI(131) from 1-6 days: an increasing PBI(131) correlates with a positive scan, and a decreasing PBI(131) with a negative scan. The area under the curve (AUC) for the PBI(131)-time curve is related to the absorbed dose for the tumour. For a small group of 11 patients, dosimetry estimates were made from serial scans, quantified with phantoms; these absorbed doses correlated with the AUC and the 6-day PBI(131). Therefore, it is suggested that these parameters may be useful in predicting absorbed radiation dose in these patients. PMID- 11388992 TI - Changes in epidermal radiosensitivity with time associated with increased colony numbers. AB - Epidermal clonogenic cell survival and colony formation following irradiation were investigated and related to radiosensitivity. A rapid in vivo/in vitro assay was developed for the quantification of colonies arising from surviving clonogenic cells in pig epidermis after irradiation. Bromodeoxyuridine (BrdU) labelled cells in full thickness epidermal sheets were visualized using standard immunohistochemistry. In unirradiated skin, approximately 900 BrdU-positive cells mm(-2) were counted. In a time sequence experiment, BrdU-positive cell numbers increased from an average of 900 cells mm(-2) to approximately 1400 cells mm(-2) after BrdU-labelling for 2-24 h. In irradiated skin, colonies containing >/=16 BrdU-positive cells were seen for the first time at days 14/15 after irradiation. The number of these colonies per cm(2) as a function of skin surface dose yielded a cell survival curve with a D(0)-value (+/-SE) of 3.9+/-0.6 Gy. This relatively high D(0)-value is possibly due to a rapid fall off in depth dose distribution for the iridium-192 source and consequently a substantial contribution of hair follicular epithelium to colony formation. At 14/15 days after irradiation, the ED(50) level of 33.6 Gy for the in vivo response of moist desquamation corresponded with 2.7 colonies cm(-2). Surprisingly, the number of colonies increased with time after irradiation with an estimated doubling time of approximately 4 days, while the D(0)-value remained virtually unchanged. This increase in colony numbers could be due to migration of clonogenic cells, to the recruitment of dormant clonogenic cell survivors by elevated levels of cytokines, or to both. Although frequent biopsying caused increased cytokine levels, which had a systemic effect on unirradiated skin, it had no influence on colony formation in irradiated skin. Smaller colonies, containing 4-8 cells or 9-15 cells, were abundant, particularly after higher doses, which resulted in higher D(0)-values. The majority of these small colonies were abortive and did not progress to larger colonies. There was no statistical evidence for significant variations in the interanimal responses. PMID- 11388993 TI - Case report: MR findings of malignant melanoma of the vagina. AB - We report a case in which malignant melanoma of the vagina showed some MR signal changes after radiotherapy. Before radiotherapy, the tumour had slightly high signal intensity on T(1) weighted images and was enhanced after gadolinium-DTPA administration. After radiotherapy, the signal intensity of the tumour increased conspicuously on both T(1) weighted images and fat suppression T(1) weighted images. PMID- 11388994 TI - Case report: Hepatic arterial occlusion following infusion catheter placement: recanalization using the Palmaz stent. AB - We report two cases in which hepatic arterial occlusion developed soon after percutaneous catheter placement for hepatic arterial infusion chemotherapy in malignant hepatic neoplasms. A replaced hepatic artery and a proper hepatic artery, respectively, became occluded in these cases. In both, because the tip of a side-holed catheter for arterial infusion was fixed in place, we performed recanalization without removing the indwelling catheter. Despite the presence of the catheter, a stent could be inserted into the occluded portion of the hepatic artery. Recanalization of the hepatic artery was achieved with stent placement, and continuation of hepatic arterial infusion became possible. PMID- 11388995 TI - Case report: Calcifying fibrous pseudotumour of the adrenal gland. AB - Calcifying fibrous pseudotumour is a distinct pathological entity usually occurring in the soft tissue of the extremities, trunk, axilla, pleura, mediastinum and peritoneum. This report describes the hitherto unreported occurrence of this tumour of the adrenal gland in a 10-year-old girl whose imaging findings closely resembled a neuroblastoma. This entity is a potential pitfall in diagnosing adrenal neuroblastoma. PMID- 11388996 TI - Case report: Hypoechoic area as an ultrasound finding suggesting subserosal invasion in polypoid carcinoma of the gall bladder. AB - Ultrasound studies have demonstrated the gall bladder wall to consist of two or more (usually three) layers. Gall bladder tumours with neither a thinned nor split outermost hyperechoic layer are confined to the mucosa or muscle layer, or have only invaded the shallow subserosa. Differentiation of these two types of tumour has posed difficulties. We report a case of polypoid gall bladder carcinoma invading the shallow subserosa with a thickened outermost hyperechoic layer but a hypoechoic area in the deeper part of the tumour. Ultrasound and pathological correlation in this case demonstrated that the hypoechoic area represented an adenocarcinoma invading the subserosa accompanied by abundant fibrosis. PMID- 11388997 TI - Pictorial review: Trigeminal nerve: anatomy and pathology. AB - MRI is the imaging modality of choice when trigeminal nerve pathology is suspected. Most lesions are readily recognizable if appropriate imaging sequences are performed. Routine cranial MRI sequences augmented by a three-dimensional gradient echo sequence such as FISP (fast inflow with steady-state precession) are sufficient to demonstrate most pathological processes involving the trigeminal nerve and nucleus. Intravenous gadolinium-DTPA occasionally provides additional diagnostic information. MRI is particularly useful in planning the management of those conditions where surgical or medical intervention can result in improvement or resolution of symptoms. In this review, examples of a range of pathologies involving the trigeminal nerve and nucleus are presented. PMID- 11388998 TI - Spastic quadriparesis in a dwarf. PMID- 11388999 TI - Treatment of a sternotomy-related false aneurysm using a covered stent-graft. PMID- 11389003 TI - Identification of a novel, human multilymphoid progenitor in cord blood. AB - The earliest stages of lymphoid commitment from human pluripotent hematopoietic stem cells have not been defined. A clonogenic subpopulation of CD34(+)CD38(-) cord blood cells were identified that expressed high levels of the CD7 antigen and possessed only lymphoid potential. CD34(+)CD38(-)CD7(+) (CD7(+)) cells uniformly coexpressed CD45RA and HLA-DR; c-kit and Thy-1 expression was absent to low. Clonal analysis demonstrated that single CD7(+) cells could generate B cells, natural killer cells, and dendritic cells but were devoid of myeloid or erythroid potential. In contrast, control CD34(+)CD38(-)CD7(-) (CD7(-)) cells generated both lymphoid and myelo-erythroid cells. The lymphoid potential (generation of lymphoid progeny in bulk and single cell cultures) of CD7(+) cells was equivalent to that of the pluripotent CD7(-) cells. RNA expression studies showed that CD7(+) cells expressed PU.1 and GATA-3, but did not express Pax-5, terminal deoxynucleotide transferase, or CD3epsilon. In contrast to the previously described murine common lymphoid progenitor, the alpha chain of the receptor for interleukin-7 was not detected by fluorescence-activated cell sorting analysis or RNA polymerase chain reaction in CD7(+) cells. These studies identify a clonogenic lymphoid progenitor with both B-cell and natural killer cell lineage potential with a molecular profile that suggests a developmental stage more primitive than previously identified lymphoid progenitors. The CD7(+) phenotype distinguishes primitive human lymphoid progenitors from pluripotent stem cells, thus allowing the study of regulation of early human lymphopoiesis and providing an alternative to pluripotent stem cells for genetic manipulation and transplantation. (Blood. 2001;97:3683-3690) PMID- 11389004 TI - Wound-healing defects in mice lacking fibrinogen. AB - In addition to its key role in the control of blood loss following injury, fibrin(ogen) has been proposed to play an important role in tissue repair by providing an initial matrix that can stabilize wound fields and support local cell proliferation and migration. To test directly these concepts, the effect of fibrinogen deficiency on cutaneous tissue repair in mice was investigated using incisional and excisional wounds. The time required to overtly heal wounds was similar in fibrinogen-deficient and control mice, but histologic evaluation revealed distinct differences in the repair process, including an altered pattern of epithelial cell migration and increased epithelial hyperplasia. Furthermore, granulation tissue in fibrinogen-deficient mice failed to adequately close the wound gap, resulting in persistent open wounds or partially covered sinus tracts. The tensile strength of these wounds was also reduced compared with control mice. The most profound defect in wound tissue organization was observed in fibrinogen deficient mice following the subcutaneous implantation of a porous tubing chamber. Cells migrated into the wall of the implants at a similar rate as control mice, but cells from fibrinogen-deficient animals were unable to efficiently organize and migrate into wound fluid-filled dead space within the center of the implants. These studies show that re-epithelialization, granulation tissue formation, including the establishment of neovasculature, and the formation of fibrotic scar tissue can proceed in the absence of fibrin(ogen) and all of its proteolytic derivatives. However, fibrin (ogen) is important for appropriate cellular migration and organization within wound fields and in initially establishing wound strength and stability. (Blood. 2001;97:3691-3698) PMID- 11389005 TI - Short-pulse B-non-Hodgkin lymphoma-type chemotherapy is efficacious treatment for pediatric anaplastic large cell lymphoma: a report of the Berlin-Frankfurt Munster Group Trial NHL-BFM 90. AB - Anaplastic large-cell lymphoma (ALCL) accounts for approximately 10% of pediatric non-Hodgkin lymphoma (NHL). Previous experience from NHL-Berlin-Frankfurt-Munster (BFM) trials indicated that the short-pulse B-NHL-type treatment strategy may also be efficacious for ALCL. The purpose of this study was to test the efficacy of this protocol for treatment of childhood ALCL in a large prospective multicenter trial and to define risk factors. From April 1990 to March 1995, 89 patients younger than 18 years of age with newly diagnosed ALCL were enrolled in trial NHL-BFM 90. Immunophenotype was T-cell in 40 patients, B-cell in 5, null in 31, and not determined in 13. Stages were as follows: I, n = 8; II, n = 20; III, n = 55; IV, n = 6. Extranodal manifestations were as follows: mediastinum, n = 28; lung, n = 13; skin, n = 16; soft tissue, n = 13; bone, n = 14; central nervous system, n = 1; bone marrow, n = 5. After a cytoreductive prephase, treatment was stratified into 3 branches: patients in K1 (stage I and II resected) received three 5-day courses (methotrexate [MTX] 0.5 g/m(2), dexamethasone, oxazaphorins, etoposide, cytarabine, doxorubicin, and intrathecal therapy); patients in K2 (stage II nonresected and stage III) received 6 courses; patients in K3 (stage IV or multifocal bone disease) received 6 intensified courses including MTX 5 g/m(2), high-dose cytarabine/etoposide. The Kaplan-Meier estimate for a 5-year event-free survival was 76% +/- 5% (median follow-up, 5.6 years) for all patients and 100%, 73% +/- 6%, and 79% +/- 11% for K1, K2, and K3, respectively. Events were as follows: progression during therapy, n = 2; progression or relapse after therapy, n = 20; second malignancy, n = 1. It was concluded that short-pulse chemotherapy, stratified according to stage, is effective treatment for pediatric ALCL. B symptoms were associated with increased risk of failure. (Blood. 2001;97:3699-3706) PMID- 11389006 TI - Tumor necrosis factor alpha promoter polymorphisms influence the phenotypic expression of hereditary hemochromatosis. AB - Severe iron overload usually develops in patients with hereditary hemochromatosis (HHC), but variability in the phenotypic expression of the disease has been reported. This study assessed whether tumor necrosis factor alpha (TNF-alpha) plays a role in phenotypic expression of HHC. Sixty-four patients with HHC and 172 healthy volunteers (controls) were studied. Release of TNF-alpha from stimulated peripheral blood monocytes was measured by enzyme-linked immunosorbent assay, and 308 and 238 TNF-alpha polymorphisms were detected with polymerase chain reaction and restriction fragment-length polymorphism analysis. The relation between TNF-alpha polymorphisms and clinical expression of HHC was evaluated. Patients with HHC released less TNF-alpha than controls, but the difference was significant only in homozygotes for the C282Y mutation. The prevalence of the 308 TNF-alpha polymorphism was similar in patients and controls, whereas the prevalence of the 238 polymorphic allele was significantly lower in patients (3% versus 16%; P =.002). A lower prevalence of cirrhosis was observed in patients with TNF-alpha polymorphism than in those without it (4 of 15 [27%] versus 28 of 49 [57%]), but the difference was not significant (P =.07). In nonhomozygotes for the C282Y mutation, severe liver siderosis was less prevalent in patients with the 308 polymorphism than in those without it (P =.05). Alanine aminotransferase (ALT) values were significantly lower in patients with TNF-alpha polymorphism (P =.006), even when patients with other hepatotoxic factors were excluded. Multivariate analysis showed that TNF-alpha polymorphism was independently associated with ALT values (P =.0008 and P =.045, respectively, in homozygotes and nonhomozygotes for the C282Y mutation) and siderosis in nonhomozygotes (P =.047). Thus, TNF-alpha appears to play a role in HHC by modulating the severity of liver damage. (Blood. 2001;97:3707-3712) PMID- 11389007 TI - The Burkitt-like lymphomas: a Southwest Oncology Group study delineating phenotypic, genotypic, and clinical features. AB - The Revised European-American Lymphoma classification gives Burkitt-like lymphoma (BLL) provisional status, leaving unresolved the differential diagnosis with Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL). This study compared the biologic features of adult BLL and DLBCL. The phenotypic distinction between BLL and DLBCL was determined by immunohistochemical staining of frozen tissue from 13 patients with BLL and 55 patients with DLBCL by using an extensive antibody panel including Ki-67, CD10, CD11a/lymphocyte function-associated antigen 1alpha (LFA-1alpha), CD18/LFA-1beta, CD58/LFA-3, and CD54/intercellular adhesion molecule, CD8 for tumor-infiltrating cytotoxic T cells (T-TILs), CD44 homing receptor, and p53 and Bcl-2 oncogenic proteins. Compared with DLBCL, BLL had a higher proliferative rate (mean Ki-67, 88% versus 53%), greater expression of CD10 and p53 antigens, and decreased expression of Bcl-2. BLL cases had a consistent absence of one or more cell adhesion molecules (92% versus 27%), low T TIL numbers, and absence of CD44 homing receptor (92% versus 14%). The t(8;14) translocation was identified in 80% of BLL cases, but no patients with BLL had the t(14;18) translocation. In a 10-year analysis, median survival of patients with BLL was 1.2 years, and that of patients with DLBCL was 2.5 years. Although the proportion of patients cured was similar in the 2 groups, BLL patients had an increased risk of early death. We conclude that BLL can be recognized by its combined morphologic and phenotypic features and that it represents a high-grade lymphoma much closer to BL than DLBCL. Retention of the BLL category or inclusion of BLL as a variant of BL is biologically and clinically more appropriate than absorbing the category of BLL into DLBCL. (Blood. 2001;97:3713-3720) PMID- 11389008 TI - Regulation of monocyte procoagulant activity in acute myocardial infarction: role of tissue factor and tissue factor pathway inhibitor-1. AB - In acute myocardial infarction (AMI), monocyte procoagulant activity is increased and may contribute to the risk for recurrence and other thrombotic events. This study sought to investigate the role tissue factor (TF) and tissue factor pathway inhibitor-1 (TFPI-1) in the regulation of monocyte procoagulant activity in AMI. Serial venous blood samples were obtained from 40 patients with AMI undergoing revascularization by stent placement. Twenty patients with elective stenting for stable angina served as control subjects. TF proteolytic activity was measured with spectrozyme factor Xa (FXa), TF and TFPI-1 surface expression on monocytes by flow cytometry, RNA expression in whole blood by reverse transcription polymerase chain reaction, and concentrations of plasma prothrombin fragments F(1 + 2) by immunoassay. Forty-eight hours after AMI, an increase was found in TF RNA, followed by an increase in TF surface expression by 24% +/- 4% and in plasma concentration of F(1 + 2) by 103% +/- 17% (P <.05). These changes could not be attributed to the intervention because they did not occur in the control group. TFPI-1 RNA and binding to the monocyte surface remained unchanged. FXa generation by monocytes of patients with AMI increased 53.6% +/- 9% in the presence of polyclonal antibodies to TFPI-1, indicating that cell-associated TFPI-1 inhibits monocyte TF activity. The increased monocyte procoagulant activity in AMI was caused by an up-regulation of TF that was partially inhibited by surface-bound TFPI-1. Anticoagulant therapy by direct inhibition of TF activity may, thus, be particularly effective in AMI. (Blood. 2001;97:3721-3726) PMID- 11389009 TI - Biology and outcome of childhood acute megakaryoblastic leukemia: a single institution's experience. AB - To describe the clinical and biologic features of pediatric acute megakaryoblastic leukemia (AMKL) and to identify prognostic factors, experience at St Jude Children's Research Hospital was reviewed. Of 281 patients with acute myeloid leukemia treated over a 14-year period, 41 (14.6%) had a diagnosis of AMKL. Six patients had Down syndrome and AMKL, 6 had secondary AMKL, and 29 had de novo AMKL. The median age of the 22 boys and 19 girls was 23.9 months (range, 6.7-208.9 months). The rate of remission induction was 60.5%, with a 48% rate of subsequent relapse. Patients with Down syndrome had a significantly higher 2-year event-free survival (EFS) estimate (83%) than did other patients with de novo AMKL (14%) or with secondary AMKL (20%; P < or =.038). Among patients who had de novo AMKL without Down syndrome, 2-year EFS was significantly higher after allogeneic bone marrow transplantation (26%) than after chemotherapy alone (0%; P =.019) and significantly higher when performed during remission (46%) than when performed during persistent disease (0%; P =.019). The 5-year survival estimates were significantly lower for de novo AMKL (10%) than for other forms of de novo AML (42%; P <.001). Treatment outcome is very poor for patients with AMKL in the absence of Down syndrome. Remission induction is the most important prognostic factor. Allogeneic transplantation during remission offers the best chance of cure; in the absence of remission, transplantation offers no advantage over chemotherapy alone. (Blood. 2001;97:3727-3732) PMID- 11389010 TI - Factors influencing the development of an anti-factor IX (FIX) immune response following administration of adeno-associated virus-FIX. AB - The present study sought to determine the impact of the route of administration of an adeno-associated virus (AAV) vector encoding human factor IX (hFIX) on the induction of an immune response against the vector and its xenogenic transgene product, hFIX. Increasing doses of AAV-hFIX were administered by different routes to C57Bl/6 mice, which typically demonstrate significant immune tolerance to hFIX. The route of delivery had a profound impact on serum hFIX levels as well as the induction of an anti-hFIX humoral immune response. At all dose levels tested, delivery of AAV-hFIX by an intramuscular (IM) route induced an antibody response against the human FIX protein and no hFIX was detected in the serum of animals even at doses of 2 x 10(11) DNA viral particles (vp) of AAV-hFIX. This was in stark contrast to the mice that received AAV-hFIX by intraportal vein (IPV) administration. No anti-hFIX inhibitors were observed in any of these mice and therapeutic levels of hFIX were detected in the serum of all mice that received doses of 2 x 10(10) vp AAV-hFIX and higher. When pre-existing neutralizing immunity to AAV was established in mice, AAV-hFIX administration by either the IM or IPV routes did not result in detectable serum hFIX. Although hFIX expression was not observed in mice with pre-existing neutralizing immunity to AAV, an anti hFIX response was induced in all of the animals that received AAV-hFIX by the IM route. This was not observed in the preimmune mice that received AAV-hFIX by IPV administration. These results suggest that the threshold of inducing an immune response against a secreted transgene product, in this case the xenoprotein hFIX, is lower when the vector is administered by the IM route even in animals with pre existing immunity to AAV. (Blood. 2001;97:3733-3737) PMID- 11389011 TI - Variable correction of host defense following gene transfer and bone marrow transplantation in murine X-linked chronic granulomatous disease. AB - Chronic granulomatous disease (CGD) is an inherited immunodeficiency in which the absence of the phagocyte superoxide-generating nicotinamide adenine dinucleotide phosphate (NADPH) oxidase results in recurrent bacterial and fungal infections. A murine model of X-linked CGD (X-CGD) was used to explore variables influencing reconstitution of host defense following bone marrow transplantation and retroviral-mediated gene transfer. The outcomes of experimental infection with Aspergillus fumigatus, Staphylococcus aureus, or Burkholderia cepacia were compared in wild-type, X-CGD mice, and transplanted X-CGD mice that were chimeric for either wild-type neutrophils or neutrophils with partial correction of NADPH oxidase activity after retroviral-mediated gene transfer. Host defense to these pathogens was improved in X-CGD mice even with correction of a limited number of neutrophils. However, intact protection against bacterial pathogens required relatively greater numbers of oxidant-generating phagocytes compared to protection against A fumigatus. The host response also appeared to be influenced by the relative level of cellular NADPH oxidase activity, particularly for A fumigatus. These results may have implications for developing effective approaches for gene therapy of CGD. (Blood. 2001;97:3738-3745) PMID- 11389012 TI - Live attenuated measles virus induces regression of human lymphoma xenografts in immunodeficient mice. AB - Derivatives of the Edmonston-B strain of measles virus (MV-Ed) are safe, live attenuated measles virus (MV) vaccines that have been used worldwide for more than 30 years. The cytoreductive potential of MV-Ed has been investigated in murine models of both aggressive and indolent B-cell lymphoma in severe combined immunodeficient (SCID) mice. The rationale for these studies was generated by experience with viral fusogenic membrane glycoproteins as cytotoxic genes and the recognition of the potential of replicating viruses in the treatment of human malignancy. Intratumoral injection of both unmodified MV-Ed and a strain of MV-Ed genetically modified by the addition of a beta-galactosidase reporter gene (MVlacZ) induced regression of large established human lymphoma xenografts, in contrast to control therapy with UV-inactivated virus, in which all tumors progressed. The antitumor effect still occurred in the presence of passively transferred anti-MV antibody. Intravenous administration of MV also resulted in considerable slowing of tumor progression. Analysis of sections of residual tumor confirmed replication of MV within the tumors. Thus, the vaccine strain of MV mediates regression of large, established human B-cell lymphoma xenografts in SCID mice, and proof of principle is established that MV is oncolytic for lymphomas in vivo. Attenuated MVs may have value as a novel replicating-virus therapy for this group of disorders. (Blood. 2001;97:3746-3754) PMID- 11389013 TI - Development of human lymphohematopoietic stem and progenitor cells defined by expression of CD34 and CD81. AB - In this study, cord blood CD34(+) cells expressed CD81, a member of the transmembrane 4 superfamily, and were classified into 3 subpopulations on the basis of their expression levels: CD34(+)CD81(+), CD34(low)CD81(+), and CD34(+)CD81(high). The lymphohematopoietic activity of each subpopulation was then examined by using suspension and clonogenic cultures for hematopoietic potential, coculture with MS-5 cells for B-cell potential, organ cultures of thymus lobes from nonobese diabetic/severe combined immunodeficiency disease (NOD/SCID) fetal mice, coculture with stromal cells derived from NOD/SCID fetal mouse liver tissue for natural killer (NK) cell and mast cell potentials, and xenotransplantation into NOD/SCID mice for long-term repopulating (LTR) ability. CD34(+)CD81(+) cells represented a heterogeneous population that had all the lymphohematopoietic activities, including NOD/SCID mouse-repopulating ability. CD34(low)CD81(+) cells were enriched in erythroid, megakaryocytic, and NK lineage potentials but had lost T-cell and B-cell potentials and LTR ability. The CD34(+)CD81(high) fraction was depleted of most lymphohematopoietic potentials except NK cell and mast cell potentials. Thus, along the differentiation cascade from CD34(+)CD81(+) lymphohematopoietic stem cells, an up-regulation of CD81 or a down-regulation of CD34 results in a change in lymphohematopoietic properties. CD81 may serve as a marker for defining developmental stages of lymphohematopoietic stem cells. (Blood. 2001;97:3755-3762) PMID- 11389014 TI - Expression of the neuronal cyclin-dependent kinase 5 activator p35Nck5a in human monocytic cells is associated with differentiation. AB - Although cyclin-dependent kinase 5 (Cdk5) is widely expressed in human tissues, its activator p35Nck5a is generally considered to be neuron specific. In addition to neuronal cells, active Cdk5 complexes have been reported in developing tissues, such as the embryonic muscle and ocular lens, and in human leukemia HL60 cells induced to differentiate by an exposure to 1,25-dihydroxyvitamin D(3); however, its activator in these cells has not been demonstrated. The results of this study indicate that p35Nck5a is associated with Cdk5 in monocytic differentiation of hematopoietic cells. Specifically, p35Nck5a is expressed in normal human monocytes and in leukemic cells induced to differentiate toward the monocytic lineage, but not in lymphocytes or cells induced to granulocytic differentiation by retinoic acid. It is present in a complex with Cdk5 that has protein kinase activity, and when ectopically expressed together with Cdk5 in undifferentiated HL60 cells, it induces the expression of CD14 and "nonspecific" esterase, markers of monocytic phenotype. These observations not only indicate a functional relationship between Cdk5 and p35Nck5a, but also support a role for this complex in monocytic differentiation. (Blood. 2001;97:3763-3767) PMID- 11389015 TI - The adapter protein CrkL associates with CD34. AB - CD34 is a cell-surface transmembrane protein expressed specifically at the stem/progenitor stage of lymphohematopoietic development that appears to regulate adhesion. To elucidate intracellular signals modified by CD34, we designed and constructed glutathione-S-transferase (GST)- fusion proteins of the intracellular domain of full-length CD34 (GST-CD34i(full)). Precipitation of cell lysates using GST-CD34i(full) identified proteins of molecular mass 39, 36, and 33 kd that constitutively associated with CD34 and a 45-kd protein that associated with CD34 after adhesion. By Western analysis, we identified the 39-kd protein as CrkL. In vivo, CrkL was coimmunoprecipitated with CD34 using CD34 antibodies, confirming the association between CrkL and CD34. CD34 peptide inhibition assays demonstrated that CrkL interacts at a membrane-proximal region of the CD34 tail. To identify the CrkL domain responsible for interaction with CD34, we generated GST-fusion constructs of adapter proteins including GST-CrkL3' (C-terminal SH3) and GST-CrkL5' (N-terminal SH2SH3). Of these fusion proteins, only GST-CrkL3' could precipitate endogenously expressed CD34, suggesting that CD34 binds the C terminal SH3 domain of CrkL. Interestingly, there appears to be differential specificity between CrkL and CrkII for CD34, because GST-CD34i(full) did not precipitate CrkII, a highly homologous Crk family member. Furthermore, GST CD34i(full) did not bind c-Abl, c-Cbl, C3G, or paxillin proteins that are known to associate with CrkL, suggesting that CD34 directly interacts with the CrkL protein. CD34i(full) association with Grb or Shc adapter proteins was not detected. Our investigations shed new light on signaling pathways of CD34 by demonstrating that CD34 couples to the hematopoietic adapter protein CrkL. (Blood. 2001;97:3768-3775) PMID- 11389016 TI - Dimeric erythropoietin fusion protein with enhanced erythropoietic activity in vitro and in vivo. AB - High doses of recombinant human erythropoietin (rhEpo) are required for the treatment of chronic anemia. Thus, it is clear that therapy for chronic anemia would greatly benefit from an erythropoietin derivative with increased erythropoietic activity rather than the native endogenous hormone. In this report, the activity of a human Epo-Epo dimer protein, obtained by recombinant technology, is described and compared with its Epo monomer counterpart produced under identical conditions. Although monomer Epo and dimer Epo-Epo had similar pharmacokinetics in normal mice, the increase in hematocrit value was greater with the dimer than with the monomer. Moreover, in clonogenic assays using CD34(+) human hematopoietic cells, the human dimer induced a 3- to 4-fold-greater proliferation of erythroid cells than the monomer. Controlled secretion of dimeric erythropoietin was achieved in beta-thalassemic mice by in vivo intramuscular electrotransfer of a mouse Epo-Epo plasmid containing the tetO element and of a plasmid encoding the tetracycline controlled transactivator tTA. Administration of tetracycline completely inhibited the expression of the mEpo dimer. On tetracycline withdrawal, expression of the Epo-Epo dimer resumed, thereby resulting in a large and sustained hematocrit increase in beta thalassemic mice. No immunologic response against the dimer was apparent in mice because the duration of the hematocrit increase was similar to that observed with the monomeric form of mouse erythropoietin. (Blood. 2001;97:3776-3782) PMID- 11389017 TI - Autoantibody against prothrombin aberrantly alters the proenzyme to facilitate formation of a complex with its physiological inhibitor antithrombin III without thrombin conversion. AB - Acquired coagulation factor inhibitors include pathologic immunoglobulins that specifically bind to coagulation factors and either neutralize their procoagulant activity, accelerate their clearance from the circulation, or have proteolytic activity to degrade them into inactive polypeptides. Here, an autoantibody against prothrombin is described in a patient with serious hemorrhagic diatheses. The autoantibody exerts its influence by a previously unknown mechanism in which it inhibits coagulation through aberrant activation of the proenzyme in a catalytic manner. The antibody-bound prothrombin formed a stable stoichiometric complex with antithrombin III, consisting of intact prothrombin and an antithrombin III molecule cleaved at the (393)Arg-(394)Ser bond. The antibody dissociated from prothrombin after the complex formation with antithrombin III. Although the bound antibody elicited protease activity from prothrombin, the complex was not able to convert fibrinogen to fibrin or to activate protein C. Thus, this is the first description of an autoantibody that induces protease-like activity from a human proenzyme, permitting subsequent neutralization by its physiological inhibitor. (Blood. 2001;97:3783-3789) PMID- 11389018 TI - Activation of protease-activated receptors by gingipains from Porphyromonas gingivalis leads to platelet aggregation: a new trait in microbial pathogenicity. AB - The bacterium Porphyromonas gingivalis is a major etiologic agent in the pathogenesis of adult periodontitis in humans. Cysteine proteinases produced by this pathogen, termed gingipains, are considered to be important virulence factors. Among many other potentially deleterious activities, arginine-specific gingipains-R (RgpB and HRgpA) efficiently activate coagulation factors. To further expand knowledge of the interaction between gingipains and the clotting cascade, this study examined their effects on cellular components of the coagulation system. The enzymes induced an increase in intracellular calcium in human platelets at nanomolar concentrations and caused platelet aggregation with efficiency comparable to thrombin. Both effects were dependent on the proteolytic activity of the enzymes. Based on desensitization studies carried out with thrombin and peptide receptor agonists, and immunoinhibition experiments, gingipains-R appeared to be activating the protease-activated receptors, (PAR)-1 and -4, expressed on the surface of platelets. This was confirmed by the finding that HRgpA and RgpB potently activated PAR-1 and PAR-4 in transfected cells stably expressing these receptors. Cumulatively, the results indicate the existence of a novel pathway of host cell activation by bacterial proteinases through PAR cleavage. This mechanism not only represents a new trait in bacterial pathogenicity, but may also explain an emerging link between periodontitis and cardiovascular disease. (Blood. 2001;97:3790-3797) PMID- 11389019 TI - Protein kinase C-catalyzed phosphorylation of an inhibitory phosphoprotein of myosin phosphatase is involved in human platelet secretion. AB - Protein kinase C (PKC)-potentiated inhibitory phosphoprotein of myosin phosphatase (CPI) was detected in human platelets. Like smooth muscle CPI-17, in vitro phosphorylation of platelet CPI by PKC inhibited the activity of myosin phosphatase containing the PP1delta catalytic subunit and the 130-kd myosin binding subunit (MBS). Treatment of intact platelets with thrombin or the stable thromboxane A(2) analog STA(2) resulted in increased phosphorylation of both CPI and MBS at Thr-696, whereas phorbol myristate acetate (PMA) and the Ca(++) ionophore ionomycin only induced CPI phosphorylation. PMA induced slow adenosine triphosphate (ATP) secretion of fura 2-loaded platelets with no change in cytosolic Ca(++). The PMA-induced increase in CPI phosphorylation preceded phosphorylation of 20-kd myosin light chain (MLC(20)) at Ser-19 and ATP secretion. The PKC inhibitor, GF109203X, inhibited PMA-induced phosphorylation of CPI and MLC(20) with similar IC(50) values. These findings suggest that the activation of PKC by PMA induces MLC(20) phosphorylation by inhibiting myosin phosphatase through phosphorylation of CPI. STA(2)-induced MLC(20) phosphorylation was also diminished but not abolished by GF109203X, even at high concentrations that completely inhibited STA(2)-induced CPI phosphorylation. A combination of the Rho-kinase inhibitor Y-27632 and GF109203X led to a further decrease in STA(2)-induced MLC(20) phosphorylation, mainly because of a significant inhibition of MBS phosphorylation at Thr-696. Inhibition of STA(2) induced ATP release by Y-27632, GF109203X, or both appeared to correlate with the extent of MLC(20) phosphorylation. Thus, CPI phosphorylation by PKC may participate in inhibiting myosin phosphatase, in addition to the Rho-kinase mediated regulation of myosin phosphatase, during agonist-induced platelet secretion. (Blood. 2001;97:3798-3805) PMID- 11389020 TI - Differential requirements for the O-linked branching enzyme core 2 beta1-6-N glucosaminyltransferase in biosynthesis of ligands for E-selectin and P-selectin. AB - Selectins are carbohydrate-binding adhesion molecules that play important roles in control of leukocyte traffic. Glycosyltransferases involved in selectin ligand biosynthesis include the alpha1,3-fucosyltransferases FucT-VII and FucT-IV, one or more sialyltransferases, and at least one O-linked branching enzyme. Previous studies have shown that core 2 beta1-6-N-glucosaminyltransferase (C2GlcNAcT-I; EC 2.4.1.102) is required for functional modification of PSGL-1, the leukocyte P selectin ligand, but have been ambiguous on whether this enzyme is involved in E selectin ligand formation. Using an attachment and rolling assay under defined shear flow in vitro, this study shows that C2GlcNAcT-I(-) lymphoid cells stably transfected with FucT-VII complementary DNA attach and roll well on E-selectin at 1.5 dynes/cm.(2) Further, attachment and rolling on P-selectin of neutrophils is sharply reduced and that of short- term polarized Th1 cells is virtually abolished, with leukocytes from C2GlcNAcT-I(-/-) mice. In contrast, both neutrophils and Th1 cells from C2GlcNAcT-I(-/-) mice attach and roll as well as wild-type cells on E-selectin. These results show that C2GlcNAcT-I is selectively required for biosynthesis of ligands for P-selectin, but is not essential for at least some E-selectin ligands. Distinct requirements for C2GlcNAcT-I in the formation of ligands for E-selectin versus P-selectin represents a novel level of regulation of expression of selectin ligands and lymphocyte traffic. (Blood. 2001;97:3806-3811) PMID- 11389021 TI - Severe impairment of leukocyte rolling in venules of core 2 glucosaminyltransferase-deficient mice. AB - Leukocyte capture and rolling are mediated by selectins expressed on leukocytes (L-selectin) and the vascular endothelium (P- and E-selectin). To investigate the role of core 2 beta1-6-N-glucosaminyltransferase (C2GlcNAcT-I) for synthesis of functional selectin ligands in vivo, leukocyte rolling flux and velocity were studied in venules of untreated and tumor necrosis factor-alpha (TNFalpha) pretreated autoperfused cremaster muscles of C2GlcNAcT-I-deficient (core 2(-/-)) and littermate control mice. In untreated core 2(-/-) mice, leukocyte rolling was dramatically reduced with markedly increased rolling velocities (81 +/- 4 microm/s vs 44 +/- 3 microm/s). The reduced rolling in core 2(-/-) mice was due mainly to severely impaired binding of P-selectin to P-selectin glycoprotein ligand-1 (PSGL-1). Some rolling remained after blocking PSGL-1 in controls but not in core 2(-/-) mice. In TNFalpha-pretreated mice, rolling was markedly reduced in core 2(-/-) mice owing to impaired P-selectin- and E-selectin-mediated rolling. Rolling velocities in core 2(-/-) mice treated with an E-selectin blocking monoclonal antibody (59 +/- 4 microm/s) were significantly higher than in controls (14 +/- 1 microm/s), which provides further evidence for the severe impairment in P-selectin-mediated rolling. In conclusion, P-selectin ligands including PSGL-1 are largely C2GlcNAcT-I dependent. In addition, E-selectin mediated rolling in vivo is partially dependent on the targeted C2GlcNAcT-I. (Blood. 2001;97:3812-3819) PMID- 11389022 TI - The clonal analysis of anticardiolipin antibodies in a single patient with primary antiphospholipid syndrome reveals an extreme antibody heterogeneity. AB - The mechanism underlying the prothrombotic state that characterizes the primary antiphospholipid syndrome proves to be difficult to define mainly because of the variety of the phospholipid and protein targets of antiphospholipid antibodies that have been described. Much of the debate is related to the use of polyclonal antibodies during the different antiphospholipid assays. To better describe the antiphospholipid antibodies, a strategy was designed to analyze the reactivity of each one antibody making up the polyclonal anticardiolipin activity, breaking down this reactivity at the clonal level. This was performed in a single patient with primary antiphospholipid syndrome by combining (1) the antigen-specific selection of single cells sorted by flow cytometry using structurally bilayered labeled anionic phospholipids and (2) the cloning of immunoglobulin (Ig) variable (V) region genes originating from individual IgG anticardiolipin-specific B cells by a single-cell polymerase chain reaction technique. The corresponding V regions were cloned in order to express human recombinant antibodies in insect cells by a baculovirus expression system. The molecular analysis, the fine specificity, and the protein cofactor dependency of the first 5 monoclonal IgG anticardiolipins are reported here. This clonal analysis reveals the extreme heterogeneity of these antibodies, which could account for the difficulties in the previous attempts to define the pathogenic antiphospholipid response. This approach should help to unravel the complex antiphospholipid immune response and the mechanism of the prothrombotic state associated with these antibodies, but it could also shed some light on their possible origins. (Blood. 2001;97:3820-3828) PMID- 11389023 TI - Evidence for cross-talk between glycoprotein VI and Gi-coupled receptors during collagen-induced platelet aggregation. AB - Collagen-induced platelet aggregation is a complex process and involves synergistic action of integrins, immunoglobulin (Ig)-like receptors, G-protein coupled receptors and their ligands, most importantly collagen itself, thromboxane A(2) (TXA(2)), and adenosine diphosphate (ADP). The precise role of each of these receptor systems in the overall processes of activation and aggregation, however, is still poorly defined. Among the collagen receptors expressed on platelets, glycoprotein (GP) VI has been identified to play a crucial role in collagen-induced activation. GPVI is associated with the FcRgamma chain, which serves as the signal transducing unit of the receptor complex. It is well known that clustering of GPVI by highly specific agonists results in platelet activation and irreversible aggregation, but it is unclear whether collagen has the same effect on the receptor. This study shows that platelets from Galphaq-deficient mice, despite their severely impaired response to collagen, normally aggregate on clustering of GPVI, suggesting this not to be the principal mechanism by which collagen activates platelets. On the other hand, dimerization of GPVI by a monoclonal antibody (JAQ1), which by itself did not induce aggregation, provided a sufficient stimulus to potentiate platelet responses to Gi-coupled, but not Gq-coupled, agonists. The combination of JAQ1 and adrenaline or ADP, but not serotonin, resulted in alpha(IIb)beta(3)-dependent aggregation that occurred without intracellular calcium mobilization and shape change in the absence of Galphaq or the P2Y(1) receptor. Together, these results provide evidence for a cross-talk between (dimerized) GPVI and Gi-coupled receptors during collagen-induced platelet aggregation. (Blood. 2001;97:3829 3835) PMID- 11389024 TI - Role of Fc receptor gamma-chain in platelet glycoprotein Ib-mediated signaling. AB - Interaction between von Willebrand factor (vWF) and glycoprotein Ib (GPIb) stimulates tyrosine kinases and subsequent tyrosine phosphorylation events in human platelets. This study found that the combination of vWF and botrocetin, by interacting with GPIb, induced tyrosine phosphorylation of Fc receptor gamma chain (FcR gamma-chain), Syk, linker for activation of T cells (LAT), and phospholipase C gamma2 (PLCgamma2). Pretreatment of platelets with 10 microM PP1 completely inhibited these tyrosine phosphorylation events. On GPIb stimulation, Src and Lyn formed a complex with FcR gamma-chain and Syk, suggesting that Src and Lyn are involved in FcR gamma-chain tyrosine phosphorylation and downstream signals. In spite of the PLCgamma2 tyrosine phosphorylation, however, there was no intracellular calcium release and inositol 1,4,5-trisphosphate production. In Brij 35 lysates, FcR gamma-chain was found to constitutively associate with GPIb. The number of GPIb expressed on FcR gamma-chain-deficient platelets was comparable to that of the wild-type, as assessed by flow cytometry. However, tyrosine phosphorylation of Syk, LAT, and PLCgamma2 in response to vWF plus botrocetin was significantly suppressed, suggesting that FcR gamma-chain mediates activation signals related to GPIb. Compared with the aggregation response of wild-type platelets, that of FcR gamma-chain-deficient platelets in response to vWF plus botrocetin was impaired, implying that FcR gamma-chain is required for the full activation of platelets mediated by GPIb. (Blood. 2001;97:3836-3845) PMID- 11389025 TI - Immune thrombocytopenia resulting from sensitivity to metabolites of naproxen and acetaminophen. AB - In patients suspected of having drug-induced immune thrombocytopenia, antibodies reactive with normal platelets in the presence of the suspect drug can sometimes be identified, but negative results are often obtained. One reason for this is that drug metabolites, formed in vivo, can be the sensitizing agents, but very little is known about the specific metabolites that can cause this complication. Five patients were studied who developed thrombocytopenia after taking the nonsteroidal anti-inflammatory drug naproxen (3 cases) or acetaminophen (2 cases) but in whom drug-dependent antibodies could not be detected by means of the unmodified drugs. In each case, antibodies that reacted with normal target platelets in the presence of a known drug metabolite (naproxen glucuronide or acetaminophen sulfate) were identified. Four of the antibodies were specific for the glycoprotein (GP) IIb/IIIa complex, but one acetaminophen sulfate-dependent antibody reacted preferentially with GPIb/IX/V. In patients with a clinical picture suggestive of drug-induced immune thrombocytopenia, tests for metabolite dependent antibodies can be helpful in identifying the responsible agent. (Blood. 2001;97:3846-3850) PMID- 11389026 TI - Generation and biochemical analysis of human effector CD4 T cells: alterations in tyrosine phosphorylation and loss of CD3zeta expression. AB - Human effector T cells have been difficult to isolate and characterize due to their phenotypic and functional similarity to the memory subset. In this study, a biochemical approach was used to analyze human effector CD4 T cells generated in vitro by activation with anti-CD3 and autologous monocytes for 3 to 5 days. The resultant effector cells expressed the appropriate activation/differentiation markers and secreted high levels of interferon gamma (IFN-gamma) when restimulated. Biochemically, effector CD4 T cells exhibited increases in total intracellular tyrosine phosphorylation and effector-associated phosphorylated species. Paradoxically, these alterations in tyrosine phosphorylation were concomitant with greatly reduced expression of CD3zeta and CD3epsilon signaling subunits coincident with a reduction in surface T-cell receptor (TCR) expression. Because loss of CD3zeta has also been detected in T cells isolated ex vivo from individuals with cancer, chronic viral infection, and autoimmune diseases, the requirements and kinetics of CD3zeta down-regulation were examined. The loss of CD3zeta expression persisted throughout the course of effector T-cell differentiation, was reversible on removal from the activating stimulus, and was modulated by activation conditions. These biochemical changes occurred in effector T cells generated from naive or memory CD4 T-cell precursors and distinguished effector from memory T cells. The results suggest that human effector T-cell differentiation is accompanied by alterations in the TCR signal transduction and that loss of CD3zeta expression may be a feature of chronic T cell activation and effector generation in vivo. (Blood. 2001;97:3851-3859) PMID- 11389027 TI - Specific down-regulation of interleukin-12 signaling through induction of phospho STAT4 protein degradation. AB - Interleukin-12 (IL-12) plays a critical role in modulating the function of T lymphocytes and natural killer cells. IL-12 has potent antitumor effects in animal models, mediated primarily by its ability to enhance cytolytic activity and secretion of interferon-gamma (IFN-gamma). Unfortunately, the antitumor effect of IL-12 has not been demonstrated in clinical trials. Repeated administration of IL-12 in humans results in decreasing levels of IFN-gamma secretion. To understand the mechanism underlying this loss of responsiveness, the effect of IL-12 on its own signaling in activated human T cells was examined. These experiments demonstrate that the level of the signal transducer and activator of transcription 4 (STAT4) protein, a critical IL-12 signaling component, is dramatically decreased 24 hours after IL-12 stimulation, whereas levels of STAT4 messenger RNA are not affected. The decrease of STAT4 protein appears to be due to specific degradation of phospho-STAT4, possibly through the proteasome degradation pathway. Decreased levels of STAT4 protein lead to decreased STAT4 DNA-binding activity and reduced proliferation and secretion of IFN-gamma. This down-regulation of STAT4 is specific for IL-12 signaling, presumably owing to the prolonged activation of STAT4 induced by IL-12. IFN-alpha stimulation, which leads to transient phosphorylation of STAT4, does not reduce the level of STAT4 protein. These findings provide new insights into the regulation of IL-12 signaling in human T cells, where IL-12 promotes T(H)1 responses, but persistent IL-12 stimulation may also limit this response. The cellular depletion of STAT4 following prolonged IL-12 stimulation may also explain the loss of responsiveness following the repeated administration of IL-12 in clinical trials. (Blood. 2001;97:3860-3866) PMID- 11389028 TI - Cell surface receptors Ly-9 and CD84 recruit the X-linked lymphoproliferative disease gene product SAP. AB - X-linked lymphoproliferative disease (XLP) is a rare immune disorder commonly triggered by infection with Epstein-Barr virus. Major disease manifestations include fatal acute infectious mononucleosis, B-cell lymphoma, and progressive dys-gammaglobulinemia. SAP/SH2D1A, the product of the gene mutated in XLP, is a small protein that comprises a single SH2 domain and a short tail of 26 amino acids. SAP binds to a specific motif in the cytoplasmic tails of the cell surface receptors SLAM and 2B4, where it blocks recruitment of the phosphatase SHP-2. Here it is reported that Ly-9 and CD84, 2 related glycoproteins differentially expressed on hematopoietic cells, also recruit SAP. Interactions between SAP and Ly-9 or CD84 were analyzed using a novel yeast 2-hybrid system, by COS cell transfections and in lymphoid cells. Recruitment of SAP is most efficient when the specific tyrosine residues in the cytoplasmic tails of Ly-9 or CD84 are phosphorylated. It is concluded that in activated T cells, the SAP protein binds to and regulates signal transduction events initiated through the engagement of SLAM, 2B4, CD84, and Ly-9. This suggests that combinations of dysfunctional signaling pathways initiated by these 4 cell surface receptors may cause the complex phenotypes of XLP. (Blood. 2001;97:3867-3874) PMID- 11389029 TI - Comparative genomic hybridization analysis in adult T-cell leukemia/lymphoma: correlation with clinical course. AB - Sixty-four patients with adult T-cell leukemia/lymphoma (ATL; 18 patients with indolent subtype and 46 with aggressive subtype) associated with human T lymphotropic virus type 1 (HTLV-1) were analyzed using comparative genomic hybridization (CGH). The most frequent observations were gains at chromosomes 14q, 7q, and 3p and losses at chromosomes 6q and 13q. Chromosome imbalances, losses, and gains were more frequently observed in aggressive ATL than in indolent ATL, with significant differences between the 2 ATL subtypes at gains of 1q and 4q. An increased number of chromosomal imbalances was associated with a significantly shorter survival in all patients. A high number of chromosomal losses was associated with a poor prognosis in indolent ATL, whereas the presence of 7q+ was marginally associated with a good prognosis in aggressive ATL. Paired samples (ie, samples obtained at different sites from 4 patients) and sequential samples from 13 patients (from 6 during both chronic disease and acute crisis and from 7 during both acute onset and relapse) were examined by CGH and Southern blotting for HTLV-1. All but 2 paired samples showed differences on CGH assessment. Two chronic/crisis samples showed distinct results regarding both CGH and HTLV-1 integration sites, indicating clonal changes in ATL at crisis. In 11 patients, the finding of identical HTLV-1 sites and clonally related CGH results suggested a common origin of sequential samples. In contrast to chronic/crisis samples, CGH results with all acute/relapse sample pairs showed the presence of clonally related but not evolutional subclones at relapse, thereby suggesting marked chromosomal instability. In summary, clonal diversity is common during progression of ATL, and CGH alterations are associated with clinical course. (Blood. 2001;97:3875-3881) PMID- 11389030 TI - Hoechst 33342 efflux identifies a subpopulation of cytogenetically normal CD34(+)CD38(-) progenitor cells from patients with acute myeloid leukemia. AB - Efflux of Hoechst 33342 from normal hematopoietic cells identifies a "side population" (SP(+)) of negatively staining cells that, in the mouse, are largely CD34(-) and are enriched for primitive progenitors. To further characterize human SP(+) cells, blood or bone marrow from 16 patients with acute myeloid leukemia (AML) was analyzed for their presence, immunophenotype, and cytogenetic and functional properties, and for the relation between SP phenotype and multidrug resistance-1 (MDR-1) expression. The mean percentages of SP(+) and MDR(+) cells was 8.1% (range, 0.5%-29.9%) and 12.8% (range, 0%-54.8%), respectively, with no correlation between the 2 values. The percentages of SP(+) cells that were CD34(+)CD38(-), CD34(+)CD38(+), or CD34(-) were 12% (range, 0.4%-50%), 25% (range, 0.5%-96%), and 63% (range, 4%-99%). Cytogenetically abnormal cells were always detected in the SP(-)CD34(+)CD38(-) and SP(+)CD34(-) fractions, and abnormal colonies (CFC), long-term culture-initiating cells (LTC-IC), and nonobese diabetic-severe combined immunodeficiency (NOD/SCID) mouse leukemia-IC were detected in the former fraction. No progenitors were detected among SP(+)CD34(-) cells in any of these assays from 9 of 10 samples. In contrast, exclusively normal cells were detected in the SP(+)CD34(+)CD38(-) fraction from 9 of 15 samples, and CFC, LTC-IC, and multilineage engraftment in NOD/SCID mice from this subpopulation were also cytogenetically normal in 6 of 8, 6 of 7, and 2 of 2 cases studied, respectively. In contrast to murine studies, primitive progenitors are enriched among SP(+)CD34(+)CD38(-) cells from patients with AML. The molecular basis for Hoechst dye efflux is uncertain because it does not appear to be related to MDR-1 expression. (Blood. 2001;97:3882-3889) PMID- 11389031 TI - Epithelial membrane protein 2, a 4-transmembrane protein that suppresses B-cell lymphoma tumorigenicity. AB - A murine homologue of the epithelial membrane protein 2 (EMP2) gene was identified in a search for genes associated with B-cell lymphoma tumorigenicity by using suppression subtractive hybridization. Expression of EMP2 messenger RNA in primary mouse tissues was limited to certain epithelial cell types and the peritoneal lymphoid compartment. EMP2 was expressed in the poorly tumorigenic DAC B-lymphoma cell line but was significantly down-regulated in a subline selected for in vivo tumor formation in Balb/c mice. Recombinant restoration of EMP2 expression in the subline suppressed its tumorigenicity, suggesting that loss of EMP2 was a causal factor in the malignant phenotype. Recombinant overexpression of EMP2 was studied in B lymphoma and NIH3T3 cells. EMP2 in both cell types induced cell death on serum deprivation. EMP2-induced cell death correlated with the expression level of EMP2 protein and was prevented by caspase inhibitors Z VAD and Z-DEVD. These findings for the first time describe an apoptotic effect of a GAS3 family gene in lymphocytes. They also suggest that EMP2 may influence B lymphoma tumorigenicity through a functional tumor suppressor phenotype. (Blood. 2001;97:3890-3895) PMID- 11389032 TI - Coexpression of BMI-1 and EZH2 polycomb-group proteins is associated with cycling cells and degree of malignancy in B-cell non-Hodgkin lymphoma. AB - Polycomb-group (PcG) proteins, such as BMI-1 and EZH2, form multimeric gene repressing complexes involved in axial patterning, hematopoiesis, and cell cycle regulation. In addition, BMI-1 is involved in experimental lymphomagenesis. Little is known about its role in human lymphomagenesis. Here, BMI-1 and EZH2 expression patterns are analyzed in a variety of B-cell non-Hodgkin lymphomas (B NHLs), including small lymphocytic lymphoma, follicular lymphoma, large B-cell lymphoma, mantle-cell lymphoma, and Burkitt lymphoma. In contrast to the mutually exclusive pattern of BMI-1 and EZH2 in reactive follicles, the neoplastic cells in B-NHLs of intermediate- and high-grade malignancy showed strong coexpression of BMI-1 and EZH2. This pattern overlapped with the expression of Mib-1/Ki-67, a marker for proliferation. Neoplastic cells in B-NHL of low-grade malignancy were either BMI-1(low)/EZH2(+) (neoplastic centroblasts) or BMI-1(low)EZH2(-) (neoplastic centrocytes). These observations show that low-, intermediate-, and high grade B-NHLs are associated with increased coexpression of the BMI-1 and EZH2 PcG proteins, whose normal expression pattern is mutually exclusive. This expression pattern is probably caused by a failure to down-regulate BMI-1 in dividing neoplastic cells, because BMI-1 expression is absent from normal dividing B cells. These observations are in agreement with findings in studies of Bmi-1 transgenic mice. The extent of BMI-1/EZH2 coexpression correlated with clinical grade and the presence of Mib-1/Ki-67 expression, suggesting that the irregular expression of BMI-1 and EZH2 is an early event in the formation of B NHL. This points to a role for abnormal PcG expression in human lymphomagenesis. (Blood. 2001;97:3896-3901) PMID- 11389033 TI - MCL1 transgenic mice exhibit a high incidence of B-cell lymphoma manifested as a spectrum of histologic subtypes. AB - Viability-promoting genes such as BCL2 play an important role in human cancer but do not directly cause aggressive tumors. BCL2 transgenic mice develop lymphoma at low frequency, hindering studies of tumorigenesis and its inhibition in the presence of such gene products. MCL1 is a member of the BCL2 family that is highly regulated endogenously and that promotes cell viability and immortalization when introduced exogenously. Mice expressing an MCL1 transgene in hematolymphoid tissues have now been monitored for an extended period and were found to develop lymphoma with long latency and at high probability (more than 85% over 2 years). In most cases, the disease was widely disseminated and of clonal B-cell origin. A variety of histologic subtypes were seen, prominently follicular lymphoma and diffuse large-cell lymphoma. MCL1 thus sets the stage for the development of lymphoma as does BCL2, disease occurring with high probability and recapitulating a spectrum of subtypes as seen in human patients. These findings with the transgene underscore the importance of the normal, highly regulated pattern of MCL1 expression, in addition to providing a model for studying tumorigenesis and its inhibition in the presence of a viability promoting BCL2 family member. (Blood. 2001;97:3902-3909) PMID- 11389034 TI - H4(D10S170), a gene frequently rearranged in papillary thyroid carcinoma, is fused to the platelet-derived growth factor receptor beta gene in atypical chronic myeloid leukemia with t(5;10)(q33;q22). AB - The molecular cloning of the t(5;10)(q33;q22) associated with atypical chronic myeloid leukemia (CML) is reported. Fluorescence in situ hybridization (FISH), Southern blot, and reverse transcriptase- polymerase chain reaction analysis demonstrated that the translocation resulted in an H4/platelet-derived growth factor receptor betaR (PDGFbetaR) fusion transcript that incorporated 5' sequences from H4 fused in frame to 3' PDGFbetaR sequences encoding the transmembrane, WW-like, and tyrosine kinase domains. FISH combined with immunophenotype analysis showed that t(5;10)(q33;q22) was present in CD13(+) and CD14(+) cells but was not observed in CD3(+) or CD19(+) cells. H4 has previously been implicated in pathogenesis of papillary thyroid carcinoma as a fusion partner of RET. The H4/RET fusion incorporates 101 amino acids of H4, predicted to encode a leucine zipper dimerization domain, whereas the H4/PDGFbetaR fusion incorporated an additional 267 amino acids of H4. Retroviral transduction of H4/PDGFbetaR, but not a kinase-inactive mutant, conferred factor-independent growth to Ba/F3 cells and caused a T-cell lymphoblastic lymphoma in a murine bone marrow transplantation assay of transformation. Mutational analysis showed that the amino-terminal H4 leucine zipper domain (amino acids 55-93), as well as H4 amino acids 101 to 386, was required for efficient induction of factor independent growth of Ba/F3 cells. Tryptophan-to-alanine substitutions in the PDGFbetaR WW-like domain at positions 566/593, or tyrosine-to-phenylalanine substitutions at PDGFbetaR positions 579/581 impaired factor-independent growth of Ba/F3 cells. H4/PDGFbetaR is an oncoprotein expressed in t(5;10)(q33;q22) atypical CML and requires dimerization motifs in the H4 moiety, as well as residues implicated in signal transduction by PDGFbetaR, for efficient induction of factor-independent growth of Ba/F3 cells. (Blood. 2001;97:3910-3918) PMID- 11389035 TI - Angiogenesis in acute promyelocytic leukemia: induction by vascular endothelial growth factor and inhibition by all-trans retinoic acid. AB - Recent studies indicate that angiogenesis is important in the pathogenesis of leukemias, apart from its well-established role in solid tumors. In this study, the possible role of angiogenesis in acute promyelocytic leukemia (APL) was explored. Bone marrow trephine biopsies from patients with APL showed significantly increased microvessel density and hot spot density compared with normal control bone marrow biopsies. To identify the mediators of angiogenesis in APL, quantitative and functional assays were performed using the NB4 APL cell line as a model system. Conditioned media (CM) from the NB4 cells strongly stimulated endothelial cell migration. CM from the NB4 cells contained high levels of vascular endothelial growth factor (VEGF) but not basic fibroblast growth factor (bFGF). Most important, the addition of neutralizing VEGF antibodies completely inhibited the ability of NB4 CM to stimulate endothelial cell migration, suggesting that APL angiogenesis is mediated by VEGF. The effect of all-trans retinoic acid (ATRA) on APL angiogenesis was then studied. ATRA therapy resulted in a decrease in bone marrow microvessel density and hot spot density. CM from ATRA-treated APL cells did not stimulate endothelial cell migration. Finally, quantitative assays showed that ATRA treatment resulted in the abrogation of VEGF production by the NB4 cells. These results show that there is increased angiogenesis and VEGF production in APL and that ATRA therapy inhibits VEGF production and suppresses angiogenesis. The addition of specific antiangiogenic agents to differentiation therapy or chemotherapy should be explored. (Blood. 2001;97:3919-3924) PMID- 11389036 TI - Detection of leukemic cells in the CD34(+)CD38(-) bone marrow progenitor population in children with acute lymphoblastic leukemia. AB - Successful autologous hematopoietic stem cell (HSC) transplantation in childhood acute lymphoblastic leukemia (ALL) requires the ability to either selectively kill the leukemia cells or separate normal from leukemic HSC. Based on previous studies showing that more than 95% of childhood B-lineage ALL express CD38, this study evaluated whether normal CD34(+)CD38(-) progenitors from children with B lineage ALL could be isolated by flow cytometry. CD34(+) cells from bone marrow samples from 10 children with B-lineage ALL were isolated at day 28 of treatment, when clinical remission had been attained. The CD34(+) progenitor cells were flow cytometrically sorted into CD34(+)CD38(+) and CD34(+)CD38(-) populations. The absolute numbers of CD34(+)CD38(-) cells that could be isolated ranged from 401 to 6245. The cells were then analyzed for the presence of clonotypic rearrangements of the T-cell receptor (TCR) Vdelta2-Ddelta3 locus. Only patients whose diagnostic marrow had an informative TCR Vdelta2-Ddelta3 rearrangement were included in this study. Detection thresholds were typically 10(-4) to 10(-5) leukemic cells in normal marrow. In 6 of 10 samples analyzed, the sorted CD34(+)CD38(-) cells had no detectable Vdelta2-Ddelta3 rearrangements. In 4 cases, the clonotypic leukemic Vdelta2-Ddelta3 rearrangement was detected in the CD34(+)CD38(-) population, indicating that the putative normal HSC population also contained leukemic cells. The data indicate that although most childhood ALL cells express CD34 and CD38, leukemic cells are also frequently present in the CD34(+)CD38(-) population. Therefore, strategies to isolate and transplant normal HSC from children with ALL will require a more stringent definition of the normal HSC than the CD34(+)CD38(-) phenotype. (Blood. 2001;97:3925-3930) PMID- 11389037 TI - Mitochondria-targeting drugs arsenic trioxide and lonidamine bypass the resistance of TPA-differentiated leukemic cells to apoptosis. AB - Exposure of U937 human leukemic cells to the phorbol ester 12-O tetradecanoylphorbol 13-acetate (TPA) induces their differentiation into monocyte/macrophage-like cells. This terminal differentiation is associated with a resistant phenotype to apoptosis induced by the topoisomerase II inhibitor etoposide. The inhibition occurs upstream of the mitochondrial release of cytochrome c and the activation of procaspase-2, -3, -6, -7, -8, and -9. By using cell-free systems, it was demonstrated that the mitochondrial pathway to cell death that involves mitochondrial membrane depolarization, cytochrome c release and cytosolic activation of procaspases by cytochrome c/dATP remains functional in TPA-differentiated U937 cells. Accordingly, 2 drugs recently shown to target the mitochondria, namely lonidamine and arsenic trioxide, bypass the resistance of TPA-differentiated U937 cells to classical anticancer drugs. Cell death induced by the 2 compounds is associated with mitochondrial membrane depolarization, release of cytochrome c and Smac/Diablo from the mitochondria, activation of caspases, poly(ADP-ribose) polymerase cleavage and internucleosomal DNA fragmentation. Moreover, the decreased glutathione content associated with the differentiation process amplifies the ability of arsenic trioxide to activate the mitochondrial pathway to cell death. Similar results were obtained by comparing undifferentiated and TPA-differentiated human HL60 leukemic cells. These data demonstrate that mitochondria-targeting agents bypass the resistance to classical anticancer drugs induced by TPA-mediated leukemic cell differentiation. (Blood. 2001;97:3931-3940) PMID- 11389038 TI - Plasmin-induced expression of cytokines and tissue factor in human monocytes involves AP-1 and IKKbeta-mediated NF-kappaB activation. AB - It was previously shown that plasmin activates human peripheral monocytes in terms of lipid mediator release and chemotactic migration. Here it is demonstrated that plasmin induces proinflammatory cytokine release and tissue factor (TF) expression by monocytes. Plasmin 0.043 to 1.43 CTA U/mL, but not active site-blocked plasmin, triggered concentration-dependent expression of mRNA for interleukin-1alpha (IL-1alpha), IL-1beta, tumor necrosis factor-alpha (TNF alpha), and TF with maximum responses after 4 hours. Plasmin-mediated mRNA expression was inhibited in a concentration-dependent manner by the lysine analogue trans-4-(aminomethyl)cyclohexane-1-carboxylic acid (t-AMCA). Increases in mRNA levels were followed by concentration- and time-dependent release of IL 1alpha, IL-1beta and TNF-alpha and by TF expression on monocyte surfaces. Neither cytokines nor TF could be detected when monocytes were preincubated with actinomycin D or cycloheximide. Electrophoretic mobility shift assays indicated plasmin-induced activation of NF-kappaB; DNA-binding complexes were composed of p50, p65, and c-Rel, as shown by supershift experiments. Nuclear translocation of NF-kappaB/Rel proteins coincided with IkappaBalpha degradation. At variance with endotoxic lipopolysaccharide, plasmin elicited the rapid degradation of another cytoplasmic NF-kappaB inhibitor, p105. Proteolysis of NF-kappaB inhibitors was apparently due to transient activation of IkappaB kinase (IKK) beta that reached maximum activity at 1 hour after plasmin stimulation. In addition, AP-1 binding was increased in plasmin-treated monocytes, with most complexes composed of JunD, c-Fos, and FosB. These findings further substantiate the role of plasmin as a proinflammatory activator of human monocytes and reveal an important new link between the plasminogen-plasmin system and inflammation. (Blood. 2001;97:3941 3950) PMID- 11389039 TI - Human cathelicidin, hCAP-18, is processed to the antimicrobial peptide LL-37 by extracellular cleavage with proteinase 3. AB - Cathelicidins are a family of antimicrobial proteins found in the peroxidase negative granules of neutrophils. The known biologic functions reside in the C terminus, which must be cleaved from the holoprotein to become active. Bovine and porcine cathelicidins are cleaved by elastase from the azurophil granules to yield the active antimicrobial peptides. The aim of this study was to identify the physiological setting for cleavage of the only human cathelicidin, hCAP-18, to liberate the antibacterial and cytotoxic peptide LL-37 and to identify the protease responsible for this cleavage. Immunoelectron microscopy demonstrated that both hCAP-18 and azurophil granule proteins were present in the phagolysosome. Immunoblotting revealed no detectable cleavage of hCAP-18 in cells after phagocytosis. In contrast, hCAP-18 was cleaved to generate LL-37 in exocytosed material. Of the 3 known serine proteases from azurophil granules, proteinase 3 was solely responsible for cleavage of hCAP-18 after exocytosis. This is the first detailed study describing the generation of a human antimicrobial peptide from a promicrobicidal protein, and it demonstrates that the generation of active antimicrobial peptides from common proproteins occurs differently in related species. (Blood. 2001;97:3951-3959) PMID- 11389040 TI - Effects of mixed hematopoietic chimerism in a mouse model of bone marrow transplantation for sickle cell anemia. AB - Sickle cell anemia (SCA) is an inherited disorder of beta-globin, resulting in red blood cell rigidity, anemia, painful crises, organ infarctions, and reduced life expectancy. Allogeneic blood or marrow transplantation (BMT) can cure SCA but is associated with an 8% to 10% mortality rate, primarily from complications of marrow-ablative conditioning. Transplantation of allogeneic marrow after less intensive conditioning reduces toxicity but may result in stable mixed hematopoietic chimerism. The few SCA patients who inadvertently developed mixed chimerism after BMT remain symptom free, suggesting that mixed chimerism can reduce disease-related morbidity. However, because the effects of various levels of mixed chimerism on organ pathology have not been characterized, this study examined the histologic effects of an increasing percentage of normal donor hematopoiesis in a mouse model of BMT for SCA. In lethally irradiated normal mice that were reconstituted with varying ratios of T-cell-depleted marrow from normal and transgenic "sickle cell" mice, normal myeloid chimerism in excess of 25% was associated with more than 90% normal hemoglobin (Hb). However, 70% normal myeloid chimerism was required to reverse the anemia. Organ pathology, including liver infarction, was present in mice with sickle Hb (HbS) levels as low as 16.8% (19.6% normal myeloid chimerism). Histologic abnormalities increased in severity up to 80% HbS, but were less severe in mice with more than 80% HbS than in those with 40% to 80% HbS. Therefore, stable mixed chimerism resulting from nonmyeloablative BMT may reduce the morbidity from SCA, but prevention of all disease complications may require minimizing the fraction of circulating sickle red cells. (Blood. 2001;97:3960-3965) PMID- 11389041 TI - Inhibition of Plasmodium yoelii blood-stage malaria by interferon alpha through the inhibition of the production of its target cell, the reticulocyte. AB - The effect of a recombinant hybrid human interferon alpha (IFN-alpha) (which cross-reacts with murine cells) on C57BL/6 mice infected with Plasmodium yoelii sporozoites or parasitized erythrocytes was determined. IFN-alpha did not inhibit the development of the parasite in the liver, but it did reduce the blood parasite load and the hepatosplenomegaly induced by the infection in mice injected with blood-stage parasites. The extent of anemia in IFN-alpha-treated and control mice was similar, despite the lower parasite load in the IFN-alpha treated mice. The reduced blood parasite load in IFN-alpha-treated mice was associated with reduced erythropoiesis and reticulocytosis. As reticulocytes are the preferred target cells for the strain of P yoelii used (P yoelii yoelii 265 BY), it was postulated that the inhibition of reticulocytosis in IFN-alpha treated mice was causally related to the observed decreased blood parasite load. This was supported by the finding that IFN-alpha inhibited a different strain of P yoelii (17X clone A), which also displays a tropism for reticulocytes, but not a line of Plasmodium vinckei petteri, which infects only mature red blood cells. As human malaria species also display different tropism for reticulocytes, these findings could be relevant for people coinfected with multiple Plasmodium species or strains or coinfected with Plasmodium and virus. (Blood. 2001;97:3966-3971) PMID- 11389042 TI - Reduced incidence of thrombosis in mice with hereditary spherocytosis following neonatal treatment with normal hematopoietic cells. AB - Thrombosis is a life-threatening complication of hemolytic anemia in humans. Cardiac thrombi are present in all adult alpha-spectrin-deficient (sph/sph) mice with severe hereditary spherocytosis, providing a model for events preceding thrombosis. The current study evaluated (1) the timing of thrombosis initiation and (2) the effect of postnatal transplantation of normal cells on life span and thrombotic incidence in adult mice. Thrombi are detected histologically following necropsy in untreated sph/sph mice of various ages and are not observed until 6 weeks of age. Thrombotic incidence increases from 50% at 6 to 7 weeks of age to 100% at 9 weeks of age. As a potential therapy, nonablated sph/sph neonates were transfused with either genetically marked normal peripheral blood (PB), bone marrow (BM), or both and assessed for donor cells and thrombosis. A single transfusion of PB, with or without BM, significantly increases the percentage of sph/sph mice that survive to weaning (4 weeks of age). Replacement in all sph/sph recipients is limited to red blood cells (RBCs). RBCs derived from donor PB are lost within 5 weeks. PB plus BM prolongs high-level donor PB cell production better than BM alone. Thrombotic incidence is significantly reduced in all sph/sph mice treated with PB, BM, or both. Hence, the presence of normal blood cells in the peripheral circulation of neonatal and adult sph/sph mice rescues the former and abrogates the development of thrombosis in the latter. (Blood. 2001;97:3972-3975) PMID- 11389043 TI - Dipyridamole inhibits sickling-induced cation fluxes in sickle red blood cells. AB - Sickling-induced cation fluxes contribute to cellular dehydration of sickle red blood cells (SS RBCs), which in turn potentiates sickling. This study examined the inhibition by dipyridamole of the sickling-induced fluxes of Na(+), K(+), and Ca(++) in vitro. At 2% hematocrit, 10 microM dipyridamole inhibited 65% of the increase in net fluxes of Na(+) and K(+) produced by deoxygenation of SS RBCs. Sickle-induced Ca(++) influx, assayed as (45)Ca(++) uptake in quin-2-loaded SS RBCs, was also partially blocked by dipyridamole, with a dose response similar to that of Na(+) and K(+) fluxes. In addition, dipyridamole inhibited the Ca(++) activated K(+) flux (via the Gardos pathway) in SS RBCs, measured as net K(+) efflux in oxygenated cells exposed to ionophore A23187 in the presence of external Ca(++), but this effect resulted from reduced anion conductance, rather than from a direct effect on the K(+) channel. The degree of inhibition of sickling-induced fluxes was dependent on hematocrit, and up to 30% of dipyridamole was bound to RBC membranes at 2% hematocrit. RBC membrane content of dipyridamole was measured fluorometrically and correlated with sickling-induced flux inhibition at various concentrations of drug. Membrane drug content in patients taking dipyridamole for other clinical indications was similar to that producing inhibition of sickling-induced fluxes in vitro. These data suggest that dipyridamole might inhibit sickling-induced fluxes of Na(+), K(+), and Ca(++) in vivo and therefore have potential as a pharmacological agent to reduce SS RBC dehydration. (Blood. 2001;97:3976-3983) PMID- 11389044 TI - Unavailability of CD147 leads to selective erythrocyte trapping in the spleen. AB - Adhesive interactions with stromal cells and the extracellular matrix are essential for the differentiation and migration of hematopoietic progenitors. In the erythrocytic lineage, a number of adhesion molecules are expressed in the developing erythrocytes and are thought to play a role in the homing and maturation of erythrocytic progenitors. However, many of these molecules are lost during the final developmental stages leading to mature erythrocytes. One of the adhesion molecules that remains expressed in mature, circulating erythrocytes is CD147. This study shows that blockade of this molecule on the cell surface by treatment with F(ab')(2) fragments of anti-CD147 monoclonal antibody disrupts the circulation of erythrocytes, leading to their selective trapping in the spleen. Consequently, mice develop an anemia, and de novo, erythropoietin-mediated erythropoiesis in the spleen. In contrast, these changes were not seen in mice similarly treated with another antierythrocyte monoclonal antibody with a different specificity. These results suggest that the CD147 expressed on erythrocytes likely plays a critical role in the recirculation of mature erythrocytes from the spleen into the general circulation. (Blood. 2001;97:3984 3988) PMID- 11389045 TI - The snake venom toxin alboaggregin-A activates glycoprotein VI. AB - The glycoprotein (GP)-Ib-IX-V receptor complex has recently been reported to signal through a pathway similar to that used by the collagen receptor GPVI, with a critical role described for the Fc receptor gamma-chain. The evidence for this was based in part on studies with the GPIbalpha-selective snake venom toxin, alboaggregin-A. In the present study, it is reported that alboaggregin-A has activity at the collagen receptor GPVI in addition to GPIbalpha, and evidence is provided that this contributes to protein tyrosine phosphorylation, shape change, and GPIIb-IIIa-dependent aggregation. This may explain why responses to alboaggregin-A are distinct from those to von Willebrand factor-ristocetin. (Blood. 2001;97:3989-3991) PMID- 11389046 TI - Expression of macrophage inflammatory protein-3alpha, stromal cell-derived factor 1, and B-cell-attracting chemokine-1 identifies the tonsil crypt as an attractive site for B cells. AB - The expression of 3 lymphoid chemokines-macrophage inflammatory protein-3alpha (MIP-3alpha), stromal cell-derived factor-1 (SDF-1), and B-cell-attracting chemokine-1 (BCA-1)-in the tonsil and the possible correlation between their sites of expression and B-cell localization within this tissue were studied. The results show that all 3 chemokines are produced in the crypts but differ by the nature of the cells that produce them and their location within the crypt. SDF-1 and MIP-3alpha are produced by epithelial cells, but their secretion is mutually exclusive. Both MIP-3alpha- and SDF-1-expressing cells are in close contact with memory B cells. By contrast, BCA-1-producing cells in the crypt are not epithelial and form clusters colocalized with plasma cells. Altogether, these data suggest that the chemokines produced in the tonsillar crypt may (1) attract memory B cells to antigen and (2) recruit and retain plasma cells and memory B cells within the supportive epithelial microenvironment of the crypt. (Blood. 2001;97:3992-3994) PMID- 11389047 TI - Anti-CD20 monoclonal antibody for the treatment of severe, immune-mediated, pure red cell aplasia and hemolytic anemia. AB - Immune-mediated, acquired pure red cell aplasia (PRCA) is a rare disorder frequently associated with other autoimmune phenomena. Conventional immunosuppressive treatment is often unsatisfactory. Rituximab is a monoclonal antibody against the CD20 antigen, highly effective for in vivo B-cell depletion. An 18-month-old girl with both severe PRCA and autoimmune hemolytic anemia, refractory to immunosuppressive treatment, received 2 doses of rituximab, 375 mg/m(2) per week. The drug was well tolerated. After anti-CD20 therapy, substitutive treatment with intravenous immunoglobulin was started. The treatment resulted in marked depletion of B cells; a striking rise in reticulocyte count ensued, with increasing hemoglobin levels, finally leading to transfusion independence. The child is now 5 months off-therapy, with normal hemoglobin and reticulocyte levels. This case suggests a role of anti-CD20 monoclonal antibody for treatment of patients with antibody-mediated hematologic disorders. (Blood. 2001;97:3995-3997) PMID- 11389048 TI - Exploiting cancer cell cycling for selective protection of normal cells. AB - Chemotherapy of cancer is limited by its toxicity to normal cells. On the basis of discoveries in signal transduction and cell cycle regulation, novel mechanism based therapeutics are being developed. Although these cell cycle modulators were designed to target cancer cells, some of them can also be applied for a different purpose, i.e., to protect normal cells against the lethality of chemotherapy. Loss of sensitivity of cancer cells to cell cycle inhibitors can be exploited for selective protection of normal cells that retain this response. Indeed, inhibition of redundant or overactivated pathways (e.g., growth factor-activated pathways) or stimulation of absent pathways in cancer cells (e.g., p53, Rb, and p16) may not arrest cycling of cancer cells. But growth arrest of normal cells will then permit selective killing of cancer cells by cycle-dependent chemotherapy. PMID- 11389049 TI - Progressive decreases in nuclear retinoid receptors during skin squamous carcinogenesis. AB - Retinoids are essential for normal skin growth, differentiation, and apoptosis and are active pharmacologically in the prevention and treatment of skin cancers and other lesions. Retinoid effects are mediated mainly by retinoic acid receptors (RARs) and retinoid X receptors (RXRs), which act as transcription factors to alter gene expression. Using in situ hybridization, we analyzed the expression of RARs and RXRs in normal sun-exposed skin (n = 85), squamous cell carcinoma (SCC; n = 28), and actinic keratosis [AK (a precursor to SCC); n = 38]. The expressions of five receptors (RAR-alpha and -gamma and RXR-alpha, -beta, and -gamma) were moderate to very strong in normal skin, with higher expressions in spinous and granular layers than in the basal layer. RAR-beta expression was weak or absent in normal and lesion samples. All five receptors expressed in the skin were suppressed progressively from normal skin to premalignant skin (AK) to invasive skin SCC. Specific receptor decreases in lesions relative to normal skin ranged from 75% (RXR-beta) to 96% (RAR-alpha) in SCC and from 37% (RAR-gamma) to 68% (RXR-beta) in AK. The degree of suppression of RXR-alpha and RAR-gamma, the two predominant retinoid receptors in skin, was relatively less for RXR-alpha (58% versus 86%; P = 0.015) and relatively greater for RAR-gamma (37% versus 89%; P = 0.0001) between AK and SCC, suggesting that suppression of RXR-alpha may be an earlier event and expression of RAR-gamma may be a later event of multistep squamous skin carcinogenesis. Our results indicate that suppressed expression of retinoid receptors occurs early (in AK) and is associated with progression of squamous skin carcinogenesis to SCC. PMID- 11389050 TI - Molecular identification of latent precancers in histologically normal endometrium. AB - Discovery of somatically mutated cells in human tissues has been less frequent than would be predicted by in vitro mutational rates. We analyzed the PTEN tumor suppressor gene as an early marker for endometrial carcinogenesis, and we show that 43% of histologically normal premenopausal endometria contain rare glands that fail to express PTEN protein because of mutation and/or deletion. These persist between menstrual cycles. Histopathology of PTEN-null glands is initially unremarkable, but with progression, they form distinctive high-density clusters. These data are consistent with a progression model in which initial mutation is not rate limiting. PMID- 11389051 TI - A mechanism for androgen receptor-mediated prostate cancer recurrence after androgen deprivation therapy. AB - The development and growth of prostate cancer depends on the androgen receptor and its high-affinity binding of dihydrotestosterone, which derives from testosterone. Most prostate tumors regress after therapy to prevent testosterone production by the testes, but the tumors eventually recur and cause death. A critical question is whether the androgen receptor mediates recurrent tumor growth after androgen deprivation therapy. Here we report that a majority of recurrent prostate cancers express high levels of the androgen receptor and two nuclear receptor coactivators, transcriptional intermediary factor 2 and steroid receptor coactivator 1. Overexpression of these coactivators increases androgen receptor transactivation at physiological concentrations of adrenal androgen. Furthermore, we provide a molecular basis for this activation and suggest a general mechanism for recurrent prostate cancer growth. PMID- 11389052 TI - Discovery of new markers of cancer through serial analysis of gene expression: prostate stem cell antigen is overexpressed in pancreatic adenocarcinoma. AB - Serial analysis of gene expression (SAGE) can be used to quantify gene expression in human tissues. Comparison of gene expression levels in neoplastic tissues with those seen in nonneoplastic tissues can, in turn, identify novel tumor markers. Such markers are urgently needed for highly lethal cancers like pancreatic adenocarcinoma, which typically presents at an incurable, advanced stage. The results of SAGE analyses of a large number of neoplastic and nonneoplastic tissues are now available online, facilitating the rapid identification of novel tumor markers. We searched an online SAGE database to identify genes preferentially expressed in pancreatic cancers as compared with normal tissues. SAGE libraries derived from pancreatic adenocarcinomas were compared with SAGE libraries derived from nonneoplastic tissues. Three promising tags were identified. Two of these tags corresponded to genes (lipocalin and trefoil factor 2) previously shown to be overexpressed in pancreatic carcinoma, whereas the third tag corresponded to prostate stem cell antigen (PSCA), a recently discovered gene thought to be largely restricted to prostatic basal cells and prostatic adenocarcinomas. PSCA was expressed in four of the six pancreatic cancer SAGE libraries, but not in the libraries derived from normal pancreatic ductal cells. We confirmed the overexpression of the PSCA mRNA transcript in 14 of 19 pancreatic cancer cell lines by reverse transcription-PCR, and using immunohistochemistry, we demonstrated PSCA protein overexpression in 36 of 60 (60%) primary pancreatic adenocarcinomas. In 59 of 60 cases, the adjacent nonneoplastic pancreas did not label for PSCA. PSCA is a novel tumor marker for pancreatic carcinoma that has potential diagnostic and therapeutic implications. These results establish the validity of analyses of SAGE databases to identify novel tumor markers. PMID- 11389053 TI - Uterine adenocarcinoma in mice treated neonatally with genistein. AB - The developing fetus is uniquely sensitive to perturbation with estrogenic chemicals. The carcinogenic effect of prenatal exposure to diethylstilbestrol (DES) is the classic example. Because phytoestrogen use in nutritional and pharmaceutical applications for infants and children is increasing, we investigated the carcinogenic potential of genistein, a naturally occurring plant estrogen in soy, in an experimental animal model previously reported to result in a high incidence of uterine adenocarcinoma after neonatal DES exposure. Outbred female CD-1 mice were treated on days 1-5 with equivalent estrogenic doses of DES (0.001 mg/kg/day) or genistein (50 mg/kg/day). At 18 months, the incidence of uterine adenocarcinoma was 35% for genistein and 31% for DES. These data suggest that genistein is carcinogenic if exposure occurs during critical periods of differentiation. Thus, the use of soy-based infant formulas in the absence of medical necessity and the marketing of soy products designed to appeal to children should be closely examined. PMID- 11389054 TI - Roles of Akt and glycogen synthase kinase 3beta in the ultraviolet B induction of cyclooxygenase-2 transcription in human keratinocytes. AB - Ultraviolet B (UVB)-induced cyclooxygenase-2 (COX-2) expression plays an important role in UVB tumor promotion. We examined whether Akt and glycogen synthase kinase 3beta (GSK-3beta), components of the phosphatidylinositol 3' kinase pathway, are involved in UVB induction of COX-2 transcription. UVB caused Akt phosphorylation at both Thr-308 and Ser-473 that was inhibited by LY294002, a phosphatidylinositol 3'-kinase inhibitor. LY294002 also decreased the expression of endogenous COX-2 protein and a luciferase construct driven by COX-2 promoter. Similarly, UVB caused phosphorylation of GSK-3beta (Ser-9) and presumably inactivation of GSK-3beta. Inhibition of GSK-3beta by lithium induced endogenous COX-2 protein expression and COX-2 promoter activity. Finally, overexpression of a dominant-negative Akt mutant or wild-type GSK-3beta suppressed UVB-mediated induction of COX-2 promoter. These studies suggest that inactivation of GSK-3beta through activation of Akt plays an important role in the UVB induction of COX-2 transcription. PMID- 11389055 TI - Vitamin D receptor gene polymorphisms, insulin-like growth factors, and prostate cancer risk: a population-based case-control study in China. AB - Operating through the vitamin D receptor (VDR), vitamin D inhibits prostate cancer growth and increases insulin-like growth factor binding protein (IGFBP) expression, suggesting that the vitamin D and insulin-like growth factor (IGF) regulatory systems may operate together to affect prostate cancer. Among 191 newly diagnosed prostate cancer cases and 304 randomly selected population controls in Shanghai, China, we found no significant association between the BsmI or FokI VDR gene polymorphisms and prostate cancer risk. However, we found that among men with the ff FokI genotype, those in the highest tertile of plasma IGFBP 3 had a decreased risk versus those in the lowest tertile (odds ratio, 0.14; 95% confidence interval, 0.04-0.56; P(trend) < 0.01), whereas among men with the FF and Ff genotypes, IGFBP-3 was not associated with risk. Similarly, IGFBP-1 was inversely associated with prostate cancer risk only among men with the ff FokI genotype (odds ratio, 0.25; 95% confidence interval, 0.07-0.85; P(trend) = 0.02). No such FokI genotype-specific effects were observed for IGF-I or IGF-II. Our findings in a low-risk population suggest that the IGF and vitamin D regulatory systems may interact to affect prostate cancer risk. Larger studies are needed to confirm these findings and clarify the underlying mechanisms. PMID- 11389056 TI - Expression of nerve growth factor receptors and their prognostic value in human breast cancer. AB - Nerve growth factor (NGF) has been shown recently to be mitogenic for human breast cancer cells. In the present study, we have assayed the expression of NGF receptors (NGFRs: TrkA and p75) mRNAs in 363 human primary breast cancers, using real-time quantitative reverse transcription-PCR. NGFRs were found in all of the tumor biopsies. TrkA and p75 were positively correlated and were respectively associated with the histoprognostic grading and the tumor type. NGFRs were both related to progesterone receptors. In univariate analyses, TrkA (>upper quartile) was associated with longer overall survival. Histoprognostic grading, tumor size, node involvement, and steroid receptors were also prognostic factors. In Cox multivariate analyses, TrkA was not a prognostic parameter. This study demonstrates the expression of NGFRs in breast cancer and points out that patients with high levels of TrkA have a more favorable overall survival prognosis. PMID- 11389057 TI - Kinetics and viability of circulating endothelial cells as surrogate angiogenesis marker in an animal model of human lymphoma. AB - Circulating endothelial cells (CECs) were evaluated by flow cytometry in immunodeficient mice bearing human lymphoma. A trend toward higher CEC values was observed on days 7 and 14 after transplant, and differences versus controls were highly significant on day 21 (P = 0.0061). A strong correlation was found between CEC and tumor volume (r, 0.942; P = 0.004) and between CEC and tumor-generated VEGF (r, 0.669; P = 0.02). In mice given cyclophosphamide, most of the circulating apoptotic cells were hematopoietic and not endothelial. Conversely, in mice given endostatin, all of the increase in apoptotic cells was in the endothelial cell compartment. CEC evaluation is promising as a noninvasive, surrogate angiogenesis marker. PMID- 11389058 TI - Genetic alterations of candidate tumor suppressor ING1 in human esophageal squamous cell cancer. AB - Overexpression of ING1, a candidate tumor suppressor gene, efficiently blocks cell growth or induces apoptosis in different experimental systems. ING1 maps to chromosome 13q33-34, and because loss of the terminal region of chromosome 13q has been implicated in esophageal squamous cell cancer (ESCC), we examined ESCC for genetic alterations of ING1. Among 31 informative cases of ESCC, 58.9% of the tumors showed allelic loss at chromosome 13q33-34, and we detected four tumor specific missense nucleotide changes. These alterations were found within the PHD finger domain and nuclear localization motif of the ING1 and may be functionally involved in the development of ESCC. Because immunohistochemical study revealed that all of the ESCC samples showed loss of ING1 protein expression, genetic or epigenetic alterations that abrogate the normal function of ING1 may contribute to esophageal squamous cell carcinogenesis. PMID- 11389059 TI - p53 and K-ras mutations in lung cancers from former and never-smoking women. AB - Somatic p53 mutations are common in lung cancer. Active cigarette smoking is positively correlated with the total frequency of p53 mutations and G:C to T:A transversions on the nontranscribed (DNA coding) strand. Mutational hotspots within the p53 gene, e.g., codon 157, have been identified for tobacco-related lung cancer, whereas these same mutations are found rarely in other cancers. Such data implicate specific p53 mutations as molecular markers of smoking. Because limited data exist concerning the p53 mutation frequency and spectra in ex smokers and nonsmokers, we have analyzed p53 and K-ras mutations in 126 lung cancers from a population-based case-control study of nonsmoking (n = 117) or ex smoking (n = 9) women from Missouri with quantitative assessments of exposure to environmental tobacco smoke. Mutations in the p53 gene were found in lung cancers from lifetime nonsmokers (19%) and ex-smokers (67%; odds ratio, 9.08; 95% confidence interval, 2.06-39.98). All deletions were found in tumors from patients who were either ex-smokers or nonsmokers exposed to passive smoking. The G:C to A:T transitions (11 of 28; 39%) were the most frequent p53 mutations found and clustered in tumors from lifetime nonsmokers without passive smoke exposure. The incidence of K-ras codon 12 or 13 mutations was 11% (14 of 115 analyzed) with no difference between long-term ex-smokers and nonsmokers. These and other results indicate that p53 mutations occur more commonly in smokers and ex-smokers than in never-smokers. Such comparisons provide additional evidence of genetic damage caused by tobacco smoke during lung carcinogenesis. PMID- 11389060 TI - Human cutaneous fatty acid-binding protein induces metastasis by up-regulating the expression of vascular endothelial growth factor gene in rat Rama 37 model cells. AB - Human cutaneous fatty acid-binding protein (C-FABP) gene is capable of inducing the metastatic phenotype when overexpressed in nonmetastatic rat Rama 37 cells. However, the mechanism of how it induces metastasis is not clear. Northern and slot blot analyses revealed that expression of the endogenous vascular endothelial growth factor (VEGF) gene was increased by 3.8-5.2-fold in the C-FABP transfected cells (pSV-CFABP-R37) and in their metastatic sublines (e.g., Met-1) when compared with that in the nonmetastatic control transfectant pSV-R37 cells generated by transfection of only plasmid DNA. Higher levels of VEGF immunoreactive protein were also secreted from the malignant C-FABP-expressing cells. Reverse transcription-PCR detected two VEGF transcript isoforms, VEGF(164) and VEGF(188), in both the nonmetastatic control transfectant pSV-R37 cells and the malignant metastatic Met-1 cells. Chick chorioallantoic membrane assays showed that the conditioned medium of the control pSV-R37 cells possessed only very weak angiogenic activity, whereas conditioned media from the metastatic C FABP transfectants and their sublines were strongly angiogenic and could be inhibited by antibodies to VEGF. Transfection of VEGF(164) cDNA in an expression vector into nonmetastatic Rama 37 cells produced a cell clone (R37-VEGF-2) that expressed high levels of VEGF. Inoculation of R37-VEGF-2 cells into syngeneic Wistar Furth rats produced metastases in a significant number (Fisher's exact test, P < 0.01) of animals (18 of 31 animals), whereas the control, vector alone transfected R37-PSV cells produced no metastases (0 of 30 animals). Immunocytochemical methods demonstrated a strong positive staining for VEGF and an increased microvessel density in the primary tumors produced from PSV-VEGF-2 cells in comparison with tumors produced from control transfectants. Immunocytochemical staining for factor VIII detected a 3.5-fold increase in microvessel density of the primary tumors produced by PSV-VEGF-2 cells when compared with that of the primary tumors developed from the control pSV-R37 cells. Therefore, we suggest that overexpression of the C-FABP gene in the original transfectants induces metastasis through up-regulation of expression of the VEGF gene in this rat Rama 37 model system, and thus VEGF may play a crucial role in this particular metastatic cascade. PMID- 11389061 TI - Oxidative stress in the absence of inflammation in a mouse model for hepatitis C virus-associated hepatocarcinogenesis. AB - The mechanism of hepatocarcinogenesis in hepatitis C virus (HCV) infection is still undefined. One possibility is the involvement of oxidative stress, which can produce genetic mutations as well as gross chromosomal alterations and contribute to cancer development. We recently showed that after a long period, the core protein of HCV induces hepatocellular carcinoma (HCC) in transgenic mice with marked hepatic steatosis but without inflammation, indicating a direct involvement of HCV in hepatocarcinogenesis. To elucidate the biochemical events before the development of HCC, we examined several parameters of oxidative stress and redox homeostasis in a mouse model of HCV-associated HCC. For young mice ages 3-12 months, there was no significant difference in the levels of hydroperoxides of phosphatidylcholine (PCOOH) and phosphatidylethanolamine in liver tissue homogenates between transgenic and nontransgenic control mice. In contrast, the PCOOH level was increased by 180% in old core gene transgenic mice > 16 months old. Concurrently, there was a significant increase in the catalase activity, and there were decreases in the levels of total and reduced glutathione in the same mice. A direct in situ determination by chemiluminescence revealed an increase in hydroperoxide products by 170% even in young transgenic mice, suggesting that hydroperoxides were overproduced but immediately removed by an activated scavenger system in young mice. Electron microscopy revealed lipofuscin granules, secondary lysosomes carrying various cytoplasmic organelles, and disruption of the double membrane structure of mitochondria, and PCR analysis disclosed a deletion in mitochondrial DNA. Interestingly, alcohol caused a marked increase in the PCOOH level in transgenic mice, suggesting synergism between alcohol and HCV in hepatocarcinogenesis. The HCV core protein thus alters the oxidant/antioxidant state in the liver in the absence of inflammation and may thereby contribute to or facilitate, at least in part, the development of HCC in HCV infection. PMID- 11389062 TI - Application of the National Cancer Institute international criteria for determination of microsatellite instability in ovarian cancer. AB - Recently, the National Cancer Institute (NCI) established criteria for determination of microsatellite instability (MSI) in colorectal tumors. Although the best panel of markers for ovarian tumors is not known, we evaluated epithelial ovarian cancers for MSI based on the NCI recommendations. One hundred and nine ovarian tumors were analyzed for MSI by gel analysis of paired germ-line and tumor DNA. PCR amplification was performed using the panel of five microsatellite markers recommended by the NCI (BAT25, BAT26, D5S346, D2S123, and D17S250) and nine additional markers picked based on their genomic location (NME1, D10S197, D11S904, D13S175, DXS981, DXS6800, DXS6807, AR, and D3S1611). Tumors were characterized on the basis of: high-frequency MSI (MSI-H) if two or more of the five NCI markers showed instability or there was instability at 30% or more of all markers tested; or low-frequency MSI (MSI-L) if only one of the five NCI markers showed instability or <30% of all of the markers. All of the other tumors were considered microsatellite stable. On the basis of the NCI markers, 12 (11%) tumors demonstrated MSI-H, and 8 (7%) additional tumors had MSI L. When all of the 14 markers were considered together, 13 (12%) tumors demonstrated MSI-H (based on 30% or more unstable loci), and 26 (24%) tumors had MSI-L. A single tumor identified to have MSI-H based upon all of the markers tested would have been classified as MSI-L based upon the NCI markers alone. Inclusion of an additional dinucleotide marker (NME1) to the NCI panel allowed detection of all of the tumors with MSI-H using only six markers. MSI-H occurs in approximately 12% of invasive ovarian tumors. For optimal detection of microsatellite instability in ovarian cancer, an additional marker (NME1) may be required, along with the five recommended by the NCI. PMID- 11389063 TI - Cyclooxygenase-2 expression in human gliomas: prognostic significance and molecular correlations. AB - Cyclooxygenase (COX)-2, the inducible isoform of prostaglandin H synthase, has been implicated in the growth and progression of a variety of human cancers. Although COX-2 overexpression has been observed in human gliomas, the prognostic or clinical relevance of this overexpression has not been investigated to date. In addition, no study has analyzed the relationship between COX-2 expression and other molecular alterations in gliomas. Consequently, we examined COX-2 expression by immunohistochemistry in tumor specimens from 66 patients with low- and high-grade astrocytomas and correlated the percentage of COX-2 expression with patient survival. We also analyzed the relative importance of COX-2 expression in comparison with other clinicopathological features (age and tumor grade) and other molecular alterations commonly found in gliomas (high MIB-1 level, p53 alteration, loss of retinoblastoma (Rb) protein or p16, and high bcl-2 level). Kaplan-Meier analyses demonstrated that high COX-2 expression (>50% of cells stained positive) correlated with poor survival for the study group as a whole (P < 0.0001) and for those with glioblastoma multiforme in particular (P < 0.03). Cox regression analyses demonstrated that COX-2 expression was the strongest predictor of outcome, independent of all other variables. In addition, high COX-2 expression correlated with increasing histological grade but did not correlate with positive p53 immunostaining, bcl-2 expression, loss of p16 or retinoblastoma protein expression, or high MIB-1 expression. These findings indicate that high COX-2 expression in tumor cells is associated with clinically more aggressive gliomas and is a strong predictor of poor survival. PMID- 11389064 TI - Abnormal fragile histidine triad (FHIT) expression in advanced cervical carcinoma: a poor prognostic factor. AB - The FHIT gene is a candidate tumor suppressor gene that has been implicated in the development of cervical carcinoma. We hypothesized that abnormal Fhit expression might be a poor prognostic factor for patients with cervical cancer. The tumors from 59 high-risk patients (stage II-III) were evaluated for abnormal Fhit expression by immunohistochemical staining. Abnormal Fhit expression (absent or reduced) was noted in 66% of the specimens. There was no statistical difference with respect to stage, performance status, para-aortic node metastasis, completion of therapy, grade, race, age, and HIV status between the normal and abnormal Fhit expression groups. The 3-year survival for patients whose tumors displayed normal Fhit expression versus abnormal Fhit expression was 74% versus 37%, respectively. Univariate analysis demonstrated a difference in survival that was statistically significant for age <55 years versus > or =55 years (P = 0.015), normal Fhit expression versus abnormal Fhit expression (P = 0.015), and stage II versus stage III (P = 0.033). Multivariate analysis showed that abnormal Fhit expression was a poor prognostic factor (P = 0.015). PMID- 11389065 TI - Caveolin-1 mediates testosterone-stimulated survival/clonal growth and promotes metastatic activities in prostate cancer cells. AB - Previously, we demonstrated that up-regulation of caveolin-1 (cav-1) was associated with prostate cancer metastasis, biochemical recurrence after radical prostatectomy, and androgen insensitivity. The objective of this study was to characterize the regulation of cav-1 by testosterone (T) and to test the effects of cav-1 on prostate cancer cell survival/clonal growth and metastatic activities. Our results demonstrated that T up-regulated cav-1 protein levels in part through transcriptional regulation and significantly enhanced survival of prostate cancer cell lines ABAC3 and LNCaP after serum starvation (>40% and >60% increased viability, respectively) and in an extended clonogenic assay (approximately 4-fold and 6-fold increase in colonies, respectively). Importantly, antisense cav-1 inhibited the survival effects of T in these assay systems. Modest but not high levels of adenoviral vector-mediated cav-1 expression alone also significantly increased viability (>40%) and clonal growth (10-fold increase in colonies) after serum starvation. Analysis of spontaneous metastasis in stably transfected antisense cav-1 mouse prostate cancer cell clones demonstrated reduction of spontaneous lymph node metastasis incidence (13%), spontaneous lymph node metastasis volume (46%), and experimental lung metastasis incidence (40%) compared with vector control cell clones. Surgical castration further reduced spontaneous lymph node metastasis incidence and volume (18% and 28%, respectively) in antisense cancer cell clones, but not in vector control clones. Our studies demonstrate that cav-1 is a downstream effector of T mediated prostate cancer cell survival/clonal growth and that modest levels of cav-1 can independently promote prostate cancer cell survival/clonal growth and metastatic activities. PMID- 11389066 TI - 111In-labeled 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid-lys(8) vasotocin: a new powerful radioligand for oxytocin receptor-expressing tumors. AB - We developed a radioactive ligand for tumors expressing oxytocin receptors (OTRs) by linking the chelating agent 1,4,7,10-tetraazacyclododecane-N,N',N",N"' tetraacetic acid (DOTA) to Lys(8)-vasotocin (LVT), an analogue of oxytocin with high affinity for OTRs. The new reagent (DOTA-LVT) retained high affinity for human OTRs, as proved by in vitro affinity binding to cells endogenously expressing OTRs, such as MCF7 breast carcinoma and MOG-U-V-W glioblastoma cells lines, as well as to transiently transfected COS7 cells. In in vivo experiments, DOTA-LVT carrying (111)In showed specific binding activity to OTR-positive TS/A mouse mammary tumors. The present study opens new perspectives for imaging and, possibly, therapy of OTR-positive human tumors such as breast and endometrial carcinomas, neuroblastomas, and glioblastomas. PMID- 11389067 TI - Association of CYP1B1 codon 432 mutant allele in head and neck squamous cell cancer is reflected by somatic mutations of p53 in tumor tissue. AB - Tobacco use is causally associated with head and neck squamous cell cancer (HNSCC). Here, we present the results of a case-control study that investigated the effects that the genetic variants of the cytochrome (CYP)1A1, CYP1B1, glutathione-S-transferase (GST)M1, GSTT1, and GSTP1 genes have on modifying the risk of smoking-related HNSCC. Allelisms of the CYP1A1, GSTT1, GSTM1, and GSTT1 genes alone were not associated with an increased risk. CYP1B1 codon 432 polymorphism was found to be a putative susceptibility factor in smoking-related HNSCC. The frequency of CYP1B1 polymorphism was significantly higher (P < 0.001) in the group of smoking cases when compared with smoking controls. Additionally, an odds ratio (OR) of 4.53 (2.62-7.98) was discovered when investigating smoking and nonsmoking cases for the susceptible genotype CYP1B1*2/*2, when compared with the presence of the genotype wild type. In combination with polymorphic variants of the GST genes, a synergistic-effect OR was observed. The calculated OR for the combined genotype CYP1B1*2/*2 and GSTM1*2/*2 was 12.8 (4.09-49.7). The calculated OR for the combined genotype was 13.4 (2.92-97.7) for CYP1B1*2/*2 and GSTT1*2/*2, and 24.1 (9.36-70.5) for the combination of CYP1B1*2/*2 and GSTT1-expressors. The impact of the polymorphic variants of the CYP1B1 gene on HNSCC risk is reflected by the strong association with the frequency of somatic mutations of the p53 gene. Smokers with susceptible genotype CYP1B1*2/*2 were 20 times more likely to show evidence of p53 mutations than were those with CYP1B1 wild type. Combined genotype analysis of CYP1B1 and GSTM1 or GSTT1 revealed interactive effects on the occurrence of p53 gene mutations. The results of the present study indicate that polymorphic variants of CYP1B1 relate significantly to the individual susceptibility of smokers to HNSCC. PMID- 11389068 TI - The use of the L-plastin promoter for adenoviral-mediated, tumor-specific gene expression in ovarian and bladder cancer cell lines. AB - A 2.4-kb truncated L-plastin promoter was inserted either 5' to the LacZ gene (Ad Lp-LacZ) or 5' to the cytosine deaminase (CD) gene (Ad-Lp-CD) in a replication incompetent adenoviral vector backbone. Infectivity and cytotoxicity experiments with the LacZ and CD vectors suggested that the L-plastin promoter-driven transcriptional units were expressed at much higher levels in explants of ovarian cancer cells from patients and in established ovarian or bladder cancer cell lines than they were in normal peritoneal mesothelial cells from surgical specimens, in organ cultures of normal ovarian cells, or in the established CCD minimal deviation fibroblast cell line. Control experiments showed that this difference was not attributable to the lack of infectivity of the normal peritoneal cells, the normal ovarian cells, or the minimal deviation CCD fibroblast cell line, because these cells showed expression of the LacZ reporter gene when exposed to the replication-incompetent adenoviral vector carrying the cytomegalovirus (CMV)-driven LacZ gene (Ad-CMV-LacZ). The Ovcar-5 and Skov-3 ovarian cancer cell lines exposed to the Ad-Lp-CD adenoviral vector were much more sensitive to the prodrug 5-fluorocytosine (5FC), which is converted from the 5FC prodrug into the toxic chemical 5-fluorouracil, than was the CCD minimal deviation fibroblast cell line after exposure to the same vector. A mouse xenograft model was used to show that the Ad-Lp-CD vector/5FC system could prevent engraftment of ovarian cancer cells in nude mice. Finally, injection of the Ad-Lp-CD vector into s.c. tumor nodules generated a greater reduction of the size of the tumor nodules than did injection of the Ad-CMV-LacZ vectors into tumor nodules. The Ad-Lp-CD vectors were as suppressive to tumor growth as the Ad CMV-CD vectors. These results suggest that an adenoviral vector carrying the CD gene controlled by the L-plastin promoter (Ad-Lp-CD) may be of potential value for the i.p. therapy of ovarian cancer. PMID- 11389069 TI - Antitumor effect of beta2-microglobulin in leukemic cell-bearing mice via apoptosis-inducing activity: activation of caspase-3 and nuclear factor-kappaB. AB - We have reported previously that beta2-microglobulin (beta2m) induces apoptosis in leukemic cells in vitro, and that an interaction between beta2m and HLA class I antigen induces apoptosis. Here we examined whether beta2m can induce apoptosis in leukemic cells in vivo and whether it has an antitumor effect in tumor-bearing mice. Daily administration of 50 or 250 microg of beta2m induced apoptosis and an antitumor effect on K562 leukemia cell-bearing mice in the same manner as tumor necrosis factor-alpha. In tumor tissues in beta2m-treated mice, both caspase-3 and nuclear factor-kappaB (NF-kappaB) were stained more strongly than in control mice by anti-caspase-3 and anti-NF-kappaB p65/Rel A polyclonal antibodies. We also observed the in vivo immunological effects of beta2m on lymphoid and hematopoietic organs, such as thymus, bone marrow, Peyer's patches, liver, and spleen in normal mice. Using antibodies against caspase-3 and NF-kappaB, immunohistochemical staining showed that no specific tissues were damaged or stained in normal mice. We conclude that beta2m stimulates caspase-3 and NF kappaB pathways to induce apoptosis, making it a useful approach to a new therapy for leukemia. PMID- 11389070 TI - The bisphosphonate ibandronate promotes apoptosis in MDA-MB-231 human breast cancer cells in bone metastases. AB - Bisphosphonate (BP), a specific inhibitor of osteoclasts, has been widely used as a beneficial agent for the treatment of bone metastases in patients with breast cancer. It is well recognized that BP reduces osteolysis by promoting apoptosis in osteoclasts. However, recent animal and human data suggest that BPs not only reduce osteolysis associated with metastatic breast cancer, but also decrease tumor burden in bone. The mechanisms by which tumor burden is decreased following BP administration are unknown. Here we examined the effects of the BP ibandronate on MDA-231 human breast cancer cells in bone metastases in a well-characterized animal model of bone metastasis. Ibandronate, which was administered (s.c. daily; 4 microg/mouse/day) after bone metastases were established, inhibited the progression of established osteolytic bone metastases as assessed by radiographic analysis. Histological and histomorphometrical examination revealed that ibandronate reduced osteoclastic bone resorption, with increased apoptosis in osteoclasts. Furthermore, ibandronate also significantly decreased the MDA-231 tumor burden, with increased apoptosis in MDA-231 breast cancer cells in bone metastases. In contrast, ibandronate failed to inhibit MDA-231 tumor formation with no effects on apoptosis in MDA-231 breast cancer cells in the orthotopic mammary fat pads. These data suggest that the effects of ibandronate on apoptosis in MDA-231 breast cancer cells are restricted in bone in which ibandronate selectively deposits. Consistent with these in vivo results, a relatively high concentration of ibandronate (100 microM) increased caspase-3 activity and induced DNA fragmentation in MDA-231 breast cancer cells in culture. Moreover, a caspase inhibitor, z-Val-Ala-Asp-fluoromethyl ketone, blocked ibandronate-induced DNA fragmentation in MDA-231 cells, suggesting an involvement of caspase-3 in ibandronate-induced apoptosis. Our results suggest that BP suppresses bone metastases through promotion of apoptosis in metastatic cancer cells as well as in osteoclasts. However, it still remains open whether BP has direct anticancer actions in vivo. PMID- 11389071 TI - Isotype-specific Ras.GTP-levels predict the efficacy of farnesyl transferase inhibitors against human astrocytomas regardless of Ras mutational status. AB - Previous studies have demonstrated that astrocytomas express elevated levels of activated Ras.GTP despite the absence of activating Ras mutations. Farnesyl transferase inhibitors (FTIs) exert their antitumor effect in part through inhibition of Ras-mediated signaling. SCH66336 is a potent FTI presently undergoing clinical trials in patients with solid tumors. We evaluated the efficacy of SCH66336 against a panel of eight human astrocytoma cell lines and three human astrocytoma explant xenograft models in NOD-SCID mice. SCH66336 demonstrated variable antiproliferative effects against the cell lines, with IC(50) ranging from 0.6 microM to 32.3 microM. Two of the three human glioblastoma multiforme (GBM) xenografts demonstrated substantial growth inhibition in response to SCH66336, with up to 69% growth inhibition after 21 days of treatment. Drug efficacy could be accurately predicted using a combination of the H-, K-, and N-isotype-specific Ras.GTP levels. These data indicate that the absence of Ras mutations does not preclude chemotherapeutic efficacy by FTIs, that Ras is likely a major target of FTIs regardless of Ras mutational status, and that isotype-specific Ras.GTP levels are a promising marker of drug efficacy. PMID- 11389072 TI - Osteoprotegerin inhibits osteolysis and decreases skeletal tumor burden in syngeneic and nude mouse models of experimental bone metastasis. AB - Certain malignancies, including breast cancer, frequently metastasize to bone, where the tumor cells induce osteoclasts to locally destroy bone. Osteoprotegerin (OPG), a member of the tumor necrosis factor receptor family, is a negative regulator of osteoclast differentiation, activation, and survival. We tested the ability of recombinant OPG to inhibit tumor-induced osteoclastogenesis, osteolysis, and skeletal tumor burden in two animal models. In a syngeneic model, mouse colon adenocarcinoma (Colon-26) cells were injected into the left ventricle of mice. Treatment with OPG dose-dependently decreased the number and area of radiographically evident lytic bone lesions, which, at the highest dose, were undetectable. Histologically, OPG also decreased skeletal tumor burden and tumor associated osteoclasts. In a nude mouse model, OPG treatment completely prevented radiographic osteolytic lesions caused by human MDA-MB-231 breast cancer cells. Histologically, OPG decreased skeletal tumor burden by 75% and completely eradicated MDA tumor-associated osteoclasts. In both models, OPG had no effect on metastatic tumor burden in a panel of soft tissue organs. These data indicate that OPG may be an effective therapy for preventing osteolysis and decreasing skeletal tumor burden in patients with bone metastasis. PMID- 11389073 TI - Frequent, moderate-dose cyclophosphamide administration improves the efficacy of cytochrome P-450/cytochrome P-450 reductase-based cancer gene therapy. AB - Transduction of tumor cells with a cyclophosphamide (CPA)-activating cytochrome P 450 (P450) gene provides the capacity for localized prodrug activation and greatly sensitizes solid tumors to CPA treatment in vivo. The therapeutic impact of this P450-based cancer gene therapy strategy can be substantially enhanced by cotransduction of P450 reductase, a rate-limiting component of P450-dependent intratumoral CPA activation. The present study examined the possibility of further improving P450/P450 reductase-based gene therapy by using a novel schedule of CPA administration, involving repeated CPA injection every 6 days and previously shown to have an antiangiogenic component. 9L gliosarcoma cells transduced with the CPA-activating enzyme couple P450 2B6/P450 reductase and grown s.c. in immunodeficient severe combined immunodeficient (scid) mice were repeatedly challenged with 140 mg/kg CPA every 6 days. Full tumor regression leading to eradication of six of eight tumors was observed when the tumor size at the time of initial drug treatment was approximately 400 mm(3) (approximately 1.5% of body weight). Little or no overt toxicity of the repeated CPA treatment regimen was observed. The same CPA schedule was much less effective in inducing regression of 9L tumors that were not transduced with P450/P450 reductase. Repeated CPA treatment of mice bearing large, late-stage P450/P450 reductase transduced tumors (approximately 9-16% of body weight) resulted in major (> or =95%) regression in 15 of 16 tumors, with tumor eradication observed in 2 cases. Although CPA resistance was found to emerge in the population of P450/P450 reductase-transduced tumors, this resistance primarily involved a loss of expression of the transduced P450 and/or P450 reductase gene, rather than development of intrinsic cellular resistance to the activated form of CPA. These findings demonstrate that repeated CPA treatment on a 6 day schedule can be highly effective when combined with P450/P450 reductase gene therapy and suggest that repeated transduction of tumors with prodrug-activation genes may be necessary to achieve tumor eradication and a sustained therapeutic response. PMID- 11389074 TI - Selective targeting to the hyperactive beta-catenin/T-cell factor pathway in colon cancer cells. AB - Many colon cancers suffer mutations in either the adenomatous polyposis coli or beta-catenin genes that lead to stabilization of beta-catenin and activation of downstream T-cell factor (Tcf) target genes. We have developed a novel approach targeting colon cancer cells based on their aberrant beta-catenin/Tcf signaling pathway. A recombinant adenovirus, in which an apoptosis gene fadd is under the control of the promoter containing Tcf-responsive elements, selectively and efficiently kills colon cancer cells in which the beta-catenin/Tcf pathway is hyperactivated. Our data therefore provide a conceptual proof that aberrantly activated Wnt/beta-catenin/Tcf pathways can be used to selectively target colon cancers. PMID- 11389075 TI - The c-Jun NH(2)-terminal protein kinase/AP-1 pathway is required for efficient apoptosis induced by vinblastine. AB - Vinblastine is an important antitumor agent that induces G(2)-M arrest and subsequent apoptosis in a wide variety of cell lines, but the molecular mechanisms that link mitotic arrest and apoptosis are poorly understood. The AP-1 transcription factor has been implicated in many critical cellular processes, including apoptosis, and is a major target of the c-Jun NH(2)-terminal kinase signaling pathway that is activated by vinblastine and other microtubule inhibitors. In this study we sought to determine the role of c-Jun NH(2)-terminal kinase/AP-1 in the response of KB3 carcinoma cells to vinblastine. For this purpose, we generated KB3 cell lines that stably expressed the c-Jun dominant negative deletional mutant TAM67, which lacks the NH(2)-terminal transactivation domain. KB3-TAM67 cell lines displayed normal growth kinetics and essentially unaltered basal AP-1 activity, but vinblastine-induced phosphorylation of c-Jun and activating transcription factor-2, and AP-1 activation, were strongly inhibited. KB3-TAM67 cell lines arrested normally at G(2)-M in response to vinblastine, but were significantly more resistant to the drug, exhibiting markedly delayed apoptosis and increased overall survival, relative to control cells. To investigate the underlying mechanisms, differential expression of apoptotic regulatory genes was monitored by immunoblot and cDNA microarray analysis. We found that vinblastine treatment caused down-regulation of p53 and its target p21 and up-regulation of tumor necrosis factor alpha, Bak, and several other genes in control but not in KB3-TAM67 cells, identifying these genes as putative targets of vinblastine-inducible AP-1. These results demonstrate that vinblastine-inducible AP-1 plays a destructive, proapoptotic role and may do so by regulating the expression of a specific subset of target genes that promotes efficient apoptotic cell death following mitotic arrest. PMID- 11389076 TI - Cyclic hydroxamic-acid-containing peptide 31, a potent synthetic histone deacetylase inhibitor with antitumor activity. AB - Cyclic hydroxamic-acid-containing peptide 1 (CHAP1), designed as a hybrid of trichostatin A and trapoxin, is a lead compound for the development of potent inhibitors of histone deacetylase (HDAC). In this study, we synthesized a series of CHAP derivatives and evaluated their biological activities by monitoring the potency of their inhibition of HDAC activity, their ability to augment the expression of MHC class-I molecules in B16/BL6 cells, and their effect on cell proliferation. A structure-activity relationship study using these three assay systems revealed several requirements of their structure for the strong inhibition of HDAC not only in the cell-free situation, but also in cells. When the structures of CHAP derivatives are represented as cyclo(-Asu(NHOH)-AA(2) AA(3)-Pro or Pip-)(n), where Asu(NHOH) and Pip are zeta-hydroxamide-alpha aminosuberic acid and pipecolic acid, respectively, (a) the tetrapeptide structure (n = 1) was better than the octapeptide one (n = 2); (b) AA(2) and AA(3) should be hydrophobic; and (c) the combination of amino acid chirality should be LDLD for the strongest inhibition of HDAC in cells (LDLD > LLLD, LDLL > LLDL). cyclo(-L-Asu(NHOH)-D-Tyr(Me)-L-Ile-D-Pro-) or CHAP31 was selected as one of the strongest CHAPs, and its biological activity was characterized further. CHAP31 was much more stable in the presence of cultured cells (t(1/2) > 3000 h) than trichostatin A (t(1/2) = 14.7 h) or trapoxin A (t(1/2) = 2.10 h). CHAP31 exhibited antitumor activity in C57BL x DBA/2 F(1) (BD2F(1)) mice bearing B16/BL6 tumor cells. Furthermore, CHAP31 inhibited the growth in four of five human tumor lines implanted into nude mice. These results suggest CHAP31 to be promising as a novel therapeutic agent for cancer treatment. PMID- 11389077 TI - A naturally occurring secreted human ErbB3 receptor isoform inhibits heregulin stimulated activation of ErbB2, ErbB3, and ErbB4. AB - A variety of receptor-mediated signaling pathways are controlled by both positive and negative extracellular regulators. In this study, we demonstrate that a naturally occurring secreted form of the human ErbB3 receptor, p85-soluble ErbB3 (sErbB3), is a potent negative regulator of heregulin (HRG)-stimulated ErbB2, ErbB3, and ErbB4 activation. We show that p85-sErbB3 binds to HRG with an affinity comparable to that of full-length ErbB3 and competitively inhibits high affinity HRG binding to ErbB2/ErbB3 heterodimers on the cell surface of breast carcinoma cells with an IC(50) of 0.5 nM. p85-sErbB3 inhibits HRG-induced phosphorylation of ErbB2, ErbB3, and ErbB4 in breast carcinoma-derived cell lines and can also block HRG-stimulated activation of mitogen-activated protein kinase, Akt, and association of ErbB3 with the phosphatidylinositol 3'-kinase p85 regulatory subunit. Cell growth assays show that exogenous addition of a 100-fold molar excess of p85-sErbB3 inhibits HRG-stimulated cell growth by as much as 90%. Whereas several potential mechanisms of p85-sErbB3 inhibition of ErbB receptor activation exist, our results suggest that at least one means of inhibition is competition for HRG binding. The IC(50) for both p85-sErbB3- and 2C4 (a monoclonal antibody specific for ErbB2)-mediated inhibition of HRG binding is approximately 0.5 nM, although the mechanism of inhibition by these two proteins is distinct. Together these results suggest that p85-sErbB3 is a naturally occurring negative regulator of HRG-stimulated signal transduction that may have important therapeutic applications in human malignancies associated with HRG mediated cell growth such as breast and prostate cancer. PMID- 11389078 TI - Specific localization, gamma camera imaging, and intracellular trafficking of radiolabelled chimeric anti-G(D3) ganglioside monoclonal antibody KM871 in SK-MEL 28 melanoma xenografts. AB - The chimeric monoclonal antibody KM871, directed against the G(D3) antigen, is under evaluation for its potential to target melanoma. To facilitate the in vivo evaluation of biodistribution properties and measurement of pharmacokinetics, KM871 was radiolabeled with (125)I via tyrosine residues and with (111)In via the bifunctional metal ion chelator C-functionalized trans-cyclohexyl diethylenetriaminepentaacetic acid (CHX-A"-DTPA) to lysine residues. Using antigen-positive SK-MEL-28 melanoma cells, immunoreactivities of 42 and 40% cell binding were obtained, respectively, for the two radioconjugates. Binding was enhanced in the presence of added unlabeled antibody. A humanized A33 antibody was similarly labeled with the two isotopes and used as a control. To determine and compare in vivo biodistribution characteristics of KM871 radiolabeled with (111)In or (125)I, mixtures of the radioconjugates were injected i.v. into BALB/c nude mice bearing G(D3)-positive-SK-MEL-28 melanoma xenografts. Gamma camera images were acquired; groups of five mice were sacrificed at various time intervals, and tumors, blood, and tissues were analyzed. (111)In-labeled CHX-A" DTPA-KM871 showed a maximum tumor uptake of 41.9 +/- 7.0% injected dose/g at 72 h with prolonged retention over a 15-day period. The tumor:blood ratio was 3:1 by 72 h, and higher ratios were observed at later time points. No abnormal accumulation of (111)In-labeled conjugate was found in normal tissues. In contrast, there was little accumulation of (125)I-labeled KM871 in the same tumors. The specificity of antibody localization was confirmed by the low tumor uptake values for radiolabeled control antibody. Gamma camera imaging demonstrated excellent uptake of (111)In-labeled CHX-A"-DTPA-KM871 in the xenografts. Chromatographic analyses of xenograft cytosolic extracts demonstrated tumor internalization and catabolism of radiolabeled KM871 with the formation of small molecular weight metabolites. Laser scanning confocal microscopy demonstrated that the majority of intracellular KM871 is localized to lysosomes. Despite the catabolism of the radioconjugate, a dose-dependent increase in KM871 tumor localization was shown through immunohistochemical examination of xenograft biopsies. This study demonstrates for the first time the in vivo localization of a radiolabeled anti-G(D3) monoclonal antibody to G(D3)-expressing xenografts using gamma camera scanning techniques and tumor cell internalization of KM871 tagged with a green fluorescent dye, Alexa Fluor 488, through confocal microscopy. KM871 has potential for targeting tumors in patients with melanoma. PMID- 11389079 TI - Immunophenotyping of leukemias using a cluster of differentiation antibody microarray. AB - Different leukemias express on their plasma membranes particular subsets of the 247 defined cluster of differentiation (CD) antigens, which may resemble those of precursor cells along the lineages of differentiation to mature myeloid and lymphoid leukocytes. The extent of use of CD antigen expression (immunophenotyping) for identification of leukemias has been constrained by the technique used, flow cytometry, which commonly specifies only three CD antigens in any one assay. Currently, leukemias and lymphomas are diagnosed using a combination of morphology, immunophenotype, cytochemistry, and karyotype. We have developed a rapid, simple procedure, which enables concurrent determination of 50 or more CD antigens on leukocytes or leukemia cells in a single analysis using a microarray of antibodies. A suspension of cells is applied to the array, and cells only bind to antibody dots for which they express the corresponding CD antigen. For patients with significantly raised leukocyte counts, the resulting dot pattern then represents the immunophenotype of those cells. For patients at earlier stages of disease, the diagnosis depends on recognition of dot patterns distinct from the background of normal leukocytes. Distinctive and reproducible dot patterns have been obtained for normal peripheral blood leukocytes, chronic lymphocytic leukemia (CLL), hairy cell leukemia, mantle cell lymphoma, acute myeloid leukemia, and T-cell acute lymphoblastic leukemia. The consensus pattern for CD antigen expression found on CLL cells taken from 20 patients in descending order of cells bound was CD44, HLA-DR, CD37, CD19, CD20, CD5, CD52, CD45RA, CD22, CD24, CD45, CD23, CD21, CD71, CD11c, and CD9. The antigens that provided the best discrimination between CLL and normal peripheral blood leukocytes were CD19, CD20, CD21, CD22, CD23, CD24, CD25, and CD37. Results obtained for the expression of 48 CD antigens from the microarray compared well with flow cytometry. The microarray enables extensive immunophenotyping, and the intact cells captured on antibody dots can be further characterized using soluble, fluorescently labeled antibodies. PMID- 11389081 TI - Synergy of vaccine strategies to amplify antigen-specific immune responses and antitumor effects. AB - Several different vaccine strategies have been evaluated and combined in an attempt to amplify T-cell responses toward induction of antitumor immunity. The model tumor antigen used was carcinoembryonic antigen (CEA). While initial T-cell activation studies were conducted in conventional mice, combined vaccine strategy studies and antitumor studies were conducted in transgenic mice in which CEA is expressed in normal gastrointestinal tissue and CEA protein is found in sera. The studies reported here demonstrate: (a) A recombinant avipox (fowlpox, rF) vector expressing the signal 1 (CEA) and the B7-1 costimulatory molecule transgenes (designated rF-CEA/B7-1) is more potent in inducing CEA-specific T-cell responses than rF-CEA; one administration of recombinant fowlpox vector expressing CEA and three different costimulatory molecule transgenes (B7-1, ICAM-1, LFA-3, designated rF-CEA/TRICOM) was more potent in inducing CEA-specific T-cell responses than four vaccinations with rF-CEA or two vaccinations with rF-CEA/B7 1. Moreover, up to four vaccinations with rF-CEA/TRICOM induced greater CEA specific T-cell responses with each vaccination. (b) A diversified prime and boost strategy using a prime with a recombinant vaccinia vector expressing CEA and the triad of costimulatory molecules (designated rV-CEA/TRICOM) and a boost with rF-CEA/TRICOM was more potent in inducing CEA-specific T-cell responses than the repeated use of rF-CEA/TRICOM alone. (c) The addition of granulocyte macrophage colony-stimulating factor (GM-CSF) to the rF-CEA or rF-CEA/TRICOM vaccinations via the simultaneous administration of a rF-GM-CSF vector enhanced CEA-specific T-cell responses. These strategies (TRICOM/diversified prime and boost/GM-CSF) were combined to treat CEA-expressing carcinoma liver metastases in CEA-transgenic mice; vaccination was initiated 14 days posttumor transplant. Antitumor effects in terms of survival and CD8(+) and CD4(+) responses specific for CEA were also observed in this CEA-transgenic mouse model. These studies demonstrate that the use of cytokines and diversified prime and boost regimens can be combined with the use of recombinant vectors expressing signal 1 and multiple costimulatory molecules to further amplify T-cell responses toward more effective vaccine strategies. PMID- 11389080 TI - Epidermal growth factor receptor-targeted immunophotodiagnosis and photoimmunotherapy of oral precancer in vivo. AB - Immunophotodiagnosis uses a fluorescence-labeled monoclonal antibody (MAb) that recognizes a tumor-associated antigen to image the fluorescence emitted from the fluorophore-bound MAb that has localized in the tissue. It may be used to diagnose malignant or precancerous lesions, to delineate the margins for tumor resection, or as a feedback mechanism to assess response to treatment. In oral precancer, the epidermal growth factor receptor (EGFR) is overexpressed and could be used as a marker for early detection or as a target for therapy. The goal of this study was to test an anti-EGFR MAb (C225) coupled to either the near infrared fluorescent dye N,N'-di-carboxypentyl-indodicarbocyanine-5,5'-disulfonic acid for detection or a photochemically active dye (chlorin(e6)) for therapy of early premalignancy in the hamster cheek pouch carcinogenesis model. Fluorescence levels in the carcinogen-treated tissue correlated with the histological stage of the lesions when the C225-N,N'-di-carboxypentyl-indodicarbocyanine-5,5' disulfonic acid conjugate was used but did not do so with the irrelevant conjugates. Discrete areas of clinically normal mucosa with high fluorescence (hot spots) were subsequently shown by histology to contain dysplastic areas. The best contrast between normal and carcinogen-treated cheek pouches was found at 4 8 days after injection. To test the potential of immunophotodiagnosis as a feedback modality for therapeutic intervention, experiments were conducted with the same MAb conjugated to chlorin(e6) followed by illumination to reduce expression of the EGFR by a photodynamic effect. Subsequent immunophotodiagnosis showed that this treatment led to a significant reduction in fluorescence in the carcinogen-treated cheek pouch compared with nonilluminated areas. This difference between illuminated and dark areas was not seen in the normal cheek pouch. Taken together, the results demonstrate the potential for development of immunophotodiagnosis as a diagnostic tool and as a method of monitoring response to therapy and that the EGFR may be an appropriate target in head and neck cancer. PMID- 11389082 TI - Molecular cytogenetic characterization of head and neck squamous cell carcinoma and refinement of 3q amplification. AB - We applied a combination of molecular cytogenetic methods, including comparative genomic hybridization (CGH), spectral karyotyping (SKY), and fluorescence in situ hybridization, to characterize the genetic aberrations in a panel of 11 cell lines derived from head and neck squamous cell carcinoma and 1 cell line derived from premalignant oral epithelium. CGH identified recurrent chromosomal losses at 1p, 3p, 4, 8p, 10p, and 18q; gains at 3q, 5p, 8q, 9q, and 14q; and high-level amplification at 3q13, 3q25-q26, 5q22-q23, 7q21, 8q24, 11q13-q14, 12p13, 14q24, and 20q13.1. Several recurrent translocations including t(1;13)(q10;q10), t(13;13)(q10;q10), t(14;14)(q10;q10), i(8)(q10), and i(9)(q10) and breakpoint clusters at 1p11, 1q21, 3p11, 5q11, 5q13, 6q23, 8p11, 8q11, 9p13, 9q13, 10q11, 11q13, 13q10, 14q10, and 15q10 were identified by SKY. There was a good correlation between the number of aberrations identified by CGH and SKY (r = 0.69), and the analyses were both confirmatory and complementary in their assessment of genetic aberrations. Amplification at 3q26-q27 was identified in 42% of cases. Although SKY defined the derivation of 3q gain, the precise breakpoint remained unassigned. Positional cloning efforts directed at the amplified region at 3q26-q27 identified three highly overlapping nonchimeric yeast artificial chromosome clones containing the apex of amplification. The use of these yeast artificial chromosome clones as a probe for fluorescence in situ hybridization analysis allowed a detailed characterization and quantification of the 3q amplification and refinement of unassigned SKY breakpoints. PMID- 11389083 TI - High-throughput tissue microarray analysis of 3p25 (RAF1) and 8p12 (FGFR1) copy number alterations in urinary bladder cancer. AB - Studies by comparative genomic hybridization revealed that the chromosomal regions 3p25 and 8p11-p12 are recurrently amplified in bladder cancer. To investigate the prevalence of DNA copy number alterations in these chromosomal regions and study their clinical significance, we used probes for the RAF1 (3p25) and FGFR1 (8p12) genes for fluorescence in situ hybridization. A tissue microarray containing 2317 tumors was analyzed. The analysis revealed RAF1 amplification in 4.0% and FGFR1 amplification in 3.4% of interpretable tumors. In addition, deletions were found at the 3p25 locus in 2.2% and at the 8p11-12 locus in 9.9% of interpretable tumors. Both amplifications and deletions of RAF1 and FGFR1 were significantly associated with high tumor grade (P < 0.0001), advanced stage (P < 0.0001), and poor survival (P < 0.05) if tumors of all of the stages where analyzed together. RAF1 amplifications were associated with subsequent tumor progression in pT1 carcinomas (P < 0.05). The marked differences in the frequency of all of the analyzed changes between pTa grade 1/grade 2 and pT1-4 carcinomas support the concept of these tumor groups representing different tumor entities. PMID- 11389084 TI - Increased gene amplification in immortal rodent cells deficient for the DNA dependent protein kinase catalytic subunit. AB - Gene amplification is one of the most frequent genome anomalies observed in tumor cells, whereas it has never been detected in cells of normal origin. A large body of evidence indicates that DNA double-strand breaks (DSBs) play a key role in initiating gene amplification. In mammals, DSBs are mainly repaired through the nonhomologous end-joining pathway (NHEJ) that requires a functional DNA-dependent protein kinase catalytic subunit (DNA-PKcs). In rodent cell lines, N (phosphonacetyl)-L-aspartate (PALA) resistance is considered a measure of gene amplification because it is mainly attributable to amplification of the carbamyl P-synthetase aspartate transcarbamylase dihydro-orotase (CAD) gene. In this paper we show that the radiosensitive hamster cell line V3, which is defective in DSB repair because of a mutation in the DNA-PKcs gene, displays also an increased frequency of gene amplification. In these cells, we found that the amplification of the CAD gene occurs with a frequency and a rate more than one order of magnitude higher than in control cell lines, although it relies on the same mechanisms. When the same analysis was performed in mouse embryo fibroblasts (MEFs) obtained from animals in which the DNA-PKcs gene was ablated by homologous recombination, a higher frequency of amplification compared with the controls was found only after cellular immortalization. In primary DNA-PKcs(-/-) MEFs, PALA treatment induced a block in the cell cycle, and no PALA-resistant clones were found. Our results indicate that the lack of DNA-PKcs increases the probability that gene amplification occurs in a genetic background already permissive, like that of immortalized cells, although it is not sufficient to make normal cells able to amplify. PMID- 11389085 TI - Nuclear factor-kappaB is constitutively active in C-cell carcinoma and required for RET-induced transformation. AB - Specific point mutations of the RET proto-oncogene have been demonstrated to be responsible for multiple endocrine neoplasia (MEN) types 2A and 2B, for familial medullary thyroid carcinoma (MTC) syndromes, as well as for sporadic MTC. Here we show that nuclear factor (NF)-kappaB is activated in RET-associated C-cell carcinoma specimens. TT cells, a human MTC cell line expressing MEN 2A type RET, display transcriptionally active RelA(p65) in the nucleus. NF-kappaB activity in these cells is attributable to constitutive IkappaB kinase (IKK) activity and high turn over of IkappaBalpha. RET harboring the mutations C634R (MEN 2A) or M918T (MEN 2B), in contrast to wild-type RET, activates a NF-kappaB-dependent reporter construct upon transient transfection in HeLa cells. We show that the prototype RET mutation C634R enhances phosphorylation of IkappaBalpha by IKKbeta but not by IKKalpha. RET-induced NF-kappaB and IKKbeta activity requires Ras function but does neither involve the classical mitogen-activated protein kinase kinase/extracellular signal-regulated kinase nor the phosphoinositide 3 kinase/Akt pathways. In contrast, RET-induced NF-kappaB activity is dependent on Raf and MEKK1. Inhibition of constitutive NF-kappaB activity results in cell death of TT cells and blocks focus formation induced by oncogenic forms of RET in NIH 3T3 cells. These results suggest that RET-mediated carcinogenesis critically depends on IKK activity and subsequent NF-kappaB activation. PMID- 11389086 TI - Detection of differentially expressed genes in HeLa x fibroblast hybrids using subtractive suppression hybridization. AB - To understand genetic differences and similarities between tumorigenic and nontumorigenic HeLa x fibroblast hybrid cells, subtractive suppression hybridization (SSH), based on suppression PCR and a combination of normalization and subtraction in a single procedure, was used. Using the nontumorigenic CGL1 and tumorigenic CGL3, forward (CGL1-CGL3) and reverse (CGL3-CGL1) subtracted libraries were constructed. Among 192 clones, seven were identified as differentially expressed genes specific for either CGL1 or CGL3. All seven were not reported previously as differentially expressed genes in this hybrid system. In the forward subtraction, p16 was isolated, indicating the involvement of the loss of tumorigenic phenotype. Subsequent transfection of wild-type p16 to the tumorigenic CGL3 showed growth suppression in colony formation assay; however, no tumor suppression was observed when the transfectant was inoculated into nude mice. These results indicate that: (a) SSH is a suitable method to identify differentially expressed genes in two types of cells; and (b) although p16 plays some roles in growth suppression, the p16-transfected CGL3 is still capable to proliferate in vivo. PMID- 11389087 TI - Extensive somatic microsatellite mutations in normal human tissue. AB - Microsatellite (MS) instability occurs in tumors with DNA mismatch repair (MMR) deficiencies but is typically absent in adjacent normal tissue. However, MS mutations have been observed in normal tissues from rare individuals with congenital MMR deficiencies. Autopsy tissues from a 4-year-old with congenital MMR deficiency (MLH1-/-) were examined for MS mutations. Insertions and deletions were observed in CA-repeat MS loci. Approximately 0.26 to 1.4 mutations per MS locus per cell were estimated to be present in normal heart, lymph node, kidney, and bladder epithelium. These findings illustrate that phenotypically normal MMR deficient cells commonly accumulate MS mutations. Loss of MMR and the accumulation of some MS mutations may occur early in MMR-deficient tumor progression, even before a gatekeeper mutation. PMID- 11389088 TI - Microsatellite marker analysis in screening for hereditary nonpolyposis colorectal cancer (HNPCC). AB - Hereditary nonpolyposis colorectal cancer (HNPCC) is an autosomal dominant cancer predisposition syndrome caused by germ-line mutations in DNA mismatch repair genes. It is relevant to identify HNPCC patients because colonoscopic screening of individuals with HNPCC mutations reduces cancer morbidity and mortality. Microsatellite instability (MSI) is characteristic of HNPCC tumors. A panel of five markers (BAT25, BAT26, D2S123, D5S346, and D17S250, the so-called Bethesda markers) has been proposed for screening for MSI. To test a hypothesis that the use of BAT26 alone is feasible in screening for MLH1/MSH2 mutation-positive HNPCC patients, we compared the MSI results of 494 colorectal cancer patients obtained using BAT26 with results obtained using the Bethesda markers. BAT26 was able to identify all 27 mutation-positive individuals in this series. The marker failed to identify 2 high MSI tumors and 20 low MSI tumors, all of which expressed MLH1, MSH2, and MSH6 when scrutinized by immunohistochemistry. PMID- 11389089 TI - Apoptotic triggers initiate translocations within the MLL gene involving the nonhomologous end joining repair system. AB - Translocations involving the MLL gene at 11q23 are a frequent finding in therapy related leukemia and are concentrated within a short, 8.3-kb tract of DNA, the breakpoint cluster region. In addition, a specific site adjacent to exon 12 within this region of MLL is cleaved in cells undergoing apoptosis. We show here, using human TK6 lymphoblastoid cells, that irradiation and the apoptotic trigger anti-CD95 antibody are each able to initiate translocations at the MLL exon 12 cleavage site. The translocation junctions produced contain regions of microhomology consistent with operation of the nonhomologous end joining (NHEJ) repair process. Participation of the NHEJ process is supported by the identification of the NHEJ component DNA-PKcs at the site of apoptotic cleavage. Suppression of DNA-PKcs function by the phosphatidylinositol 3-kinase inhibitor wortmannin compromises DNA end joining, increases site-specific cleavage within MLL, and eliminates MLL-restricted translocations. We propose that activation of apoptotic effector nucleases alone is sufficient to generate proleukemogenic translocations and raises the possibility that some of these may persist in cells that evade apoptotic execution and survive. PMID- 11389090 TI - Loss of expression and aberrant methylation of the CDH13 (H-cadherin) gene in breast and lung carcinomas. AB - Expression of some members of the cadherin family is reduced in several human tumors, and CDH13 (H-cadherin), located on chromosome 16q24.2-3, may function as a tumor suppressor gene. In human tumors, loss of expression of many tumor suppressor genes occurs by aberrant promoter region methylation. We examined the methylation status of the CDH13 promoter in breast and lung cancers and correlated it with mRNA expression using methylation-specific PCR and reverse transcription-PCR. Methylation was frequent in primary breast tumors (18 of 55, 33%) and cell lines (7 of 20, 35%). In lung cancers, methylation was present more frequently in non-small cell lung cancer tumors (18 of 42, 43%) and cell lines (15 of 30, 50%) than in small cell lung cancer cell lines (6 of 30, 20%; P = 0.03). Only the methylated or unmethylated forms of the gene were present in most (73 of 80, 91%) tumor cell lines. CDH13 expression was present in 24 of 30 (80%) of nonmethylated tumor lines. All 18 methylated lines tested lacked expression irrespective of whether the unmethylated form was present, confirming biallelic inactivation in methylated lines. Gene expression was restored in all five methylated cell lines tested after treatment with the demethylating agent 5'-aza 2-deoxycytidine. Our results demonstrate frequent aberrant methylation of CDH13 in breast and lung cancers accompanied by loss of gene expression, although expression may occasionally be lost by other mechanisms. PMID- 11389091 TI - Atm knock-in mice harboring an in-frame deletion corresponding to the human ATM 7636del9 common mutation exhibit a variant phenotype. AB - ATM, the gene mutated in the human immunodeficiency disorder ataxia telangiectasia (A-T), plays a central role in recognizing ionizing radiation damage in DNA and in controlling several cell cycle checkpoints. We describe here a murine model in which a nine-nucleotide in-frame deletion has been introduced into the Atm gene by homologous recombination followed by removal of the selectable marker cassette by Cre-loxP site-specific, recombination-mediated excision. This mouse, Atm-DeltaSRI, was designed as a model of one of the most common deletion mutations (7636del9) found in A-T patients. The murine Atm deletion results in the loss of three amino acid residues (SRI; 2556-2558) but produces near full-length detectable Atm protein that lacks protein kinase activity. Radiosensitivity was observed in Atm-DeltaSRI mice, whereas the immunological profile of these mice showed greater heterogeneity of T-cell subsets than observed in Atm(-/-) mice. The life span of Atm-DeltaSRI mice was significantly longer than that of Atm(-/-) mice when maintained under nonspecific pathogen-free conditions. This can be accounted for by a lower incidence of thymic lymphomas in Atm-DeltaSRI mice up to 40 weeks, after which time the animals died of other causes. The thymic lymphomas in Atm-DeltaSRI mice were characterized by extensive apoptosis, which appears to be attributable to an increased number of cells expressing Fas ligand. A variety of other tumors including B-cell lymphomas, sarcomas, and carcinomas not seen in Atm(-/-) mice were observed in older Atm-DeltaSRI animals. Thus, expression of mutant protein in Atm-DeltaSRI knock-in mice gives rise to a discernibly different phenotype to Atm(-/-) mice, which may account for the heterogeneity seen in A-T patients with different mutations. PMID- 11389092 TI - PTEN induces G(1) cell cycle arrest and decreases cyclin D3 levels in endometrial carcinoma cells. AB - Inactivating mutations in the PTEN tumor suppressor gene occur in approximately 30-50% of endometrial carcinomas. PTEN is a phosphatase that negatively regulates the phosphoinositide 3-kinase signaling pathway, including the downstream effector AKT. To evaluate the role of PTEN in endometrial growth regulation, we expressed wild-type or mutant PTEN in endometrial carcinoma cell lines. As expected, expression of exogenous PTEN decreased levels of activated AKT in all cell lines examined. However, PTEN induced a G(1) cell cycle arrest specifically in endometrial carcinoma cells that lack endogenous wild-type PTEN. Growth of cells containing wild-type PTEN was unaffected by exogenous PTEN expression. Growth arrest required a functional phosphatase domain but not the PDZ interaction motif of PTEN. Overall levels of CIP/KIP and INK4 family members, the known inhibitory regulators of the G(1) phase of the cell cycle, were unchanged. However, PTEN induced a specific reduction of cyclin D3 levels and an associated increase in the amount of the inhibitor p27(KIP1) complexed with CDK2. Enforced expression of cyclin D3 abrogated the PTEN-induced cell cycle arrest. Although PTEN signaling directly regulates p27(KIP1) levels in some settings, in endometrial carcinoma cells, PTEN expression indirectly regulated p27(KIP1) activity by modulating levels of cyclin D3. These data support multiple mechanisms of PTEN-induced cell cycle arrest. PMID- 11389093 TI - Relative expression of progesterone receptors A and B in endometrioid cancers of the endometrium. AB - The nuclear receptor for the female hormone progesterone (PR) is widely expressed in uterine cancer. PR is expressed as two proteins (PRA and PRB) with different functions, and in vitro evidence reveals PRA to inhibit PRB function, so the cellular ratio of PRA:PRB is likely to be an important determinant of progesterone action. The relative expression of PRA and B and their involvement in the pathogenesis of endometrial cancer is not known. The aims of this study were to determine PRA and B expression by dual immunofluorescent histochemistry in endometrial adenocarcinomas compared with expression in normal and hyperplastic glands, and to correlate expression in tumors with clinical features including grade. Significantly lower PR levels were found in tumors compared with normal glands and areas of complex atypical hyperplasia within the same specimen. The normal glands expressed both of the isoforms at similar levels, whereas there was increased predominance of one isoform in hyperplastic areas and in tumors, which suggested that the loss of coordinated expression of PR isoforms was an early event in tumor progression. The majority of tumors [27 (58%) of 46] expressed only one PR isoform, and the proportion expressing either PRA or B was the same [14 (30%) of 46, and 13 (28%) of 46, respectively]. One-half of all tumors ([23 (50%) of 46] expressed either PRA only or a predominance of PRA, and a few tumors [10 (22%) of 46] expressed comparable levels of PRA and B. Similar levels of PRA and B were noted only in FIGO grade 1 tumors, whereas higher grades (2 and 3) were associated with a predominance of one isoform. In summary, expression of only one PR isoform was common in endometrial cancers, which indicates that the decreased PR levels observed in these cancers arise from the loss of one PR isoform. Expression of a single PR isoform was associated with higher clinical grade, which suggests a relationship between the loss of PR isoform expression and features of poorer prognosis. Disruption of relative PR isoform expression was observed in complex atypical hyperplasia, which suggests that early alterations in the ratio of PRA:PRB may precede and/or be implicated in the development of endometrial adenocarcinoma. Alterations in the ratio of PR isoform expression are likely to cause disordered regulation of target genes, resulting in altered progestin action in the uterus, and this may be involved in the pathogenesis of endometrial cancer. PMID- 11389094 TI - Overexpression of proteins HMGA1 induces cell cycle deregulation and apoptosis in normal rat thyroid cells. AB - The high mobility group (HMG) proteins (HMGA1a, HMGA1b, and HMGA2) bind to DNA and interact with various transcriptional factors. Therefore, they play an important role in chromatin organization. HMGA protein expression is low in normal adult tissues, but abundant during embryonic development and in several experimental and human tumors. Blockage of HMGA expression inhibits the transformation of rat thyroid PC Cl 3 cells treated with oncogene-carrying retroviruses, thus implicating HMGA in rat thyroid transformation. To better understand the role of HMGA and to establish whether its up-regulated expression is sufficient to induce the transformed phenotype, we generated PC Cl 3 cells that overexpress the protein. We demonstrate that HMGA1b protein overexpression does not transform normal rat thyroid PC Cl 3 cells, but it deregulates their cell cycle: cells enter S-phase earlier and the G(2)-M transition is delayed. HMGA1-overexpressing cells undergo apoptosis through a pathway involving caspase 3 activation, probably consequent to the conflict between mitogenic pressure and the inability to proceed through the cell cycle. Using various HMGA1b gene mutations, we found that the third AT-hook domain and the acetylation site K60 are the protein regions required for induction of apoptosis in PC Cl 3 cells. In conclusion, although HMGA1 protein overexpression is associated with the malignant phenotype of rat and human thyroid cells, it does not transform normal thyroid cells in culture but leads them to programmed cell death. PMID- 11389095 TI - Protein kinase Cdelta-mediated phosphorylation of alpha6beta4 is associated with reduced integrin localization to the hemidesmosome and decreased keratinocyte attachment. AB - In mammalian epidermis, expression of the alpha6beta4 integrin is restricted to the hemidesmosome complexes, which connect the proliferative basal cell layer with the underlying basement membrane. Keratinocyte differentiation is associated with down-regulation of alpha6beta4 expression and detachment of keratinocytes from the basement membrane. Neoplastic keratinocytes delay maturation, proliferate suprabasally, and retain the expression of the alpha6beta4 integrin in suprabasal cells disassociated from the hemidesmosomes. We now show that the alpha6beta4 integrin is a substrate for serine phosphorylation by protein kinase C in keratinocytes. Furthermore, protein kinase C-mediated phosphorylation of alpha6beta4 is associated with redistribution of this integrin from the hemidesmosome to the cytosol. Specifically, in vitro kinase assays identified the protein kinase Cdelta as the primary isoform phosphorylating alpha6 and beta4 integrin subunits. Using recombinant protein kinase C adenoviruses, overexpression of protein kinase Cdelta but not protein kinase Calpha in primary keratinocytes increased beta4 serine phosphorylation, decreased alpha6beta4 localization to the hemidesmosome complexes, and reduced keratinocyte attachment. Taken together, these results establish a link between protein kinase Cdelta mediated serine phosphorylation of alpha6beta4 integrin and its effects on alpha6beta4 subcellular localization and keratinocyte attachment to the laminin underlying matrix. PMID- 11389096 TI - Effects of overexpression of gamma-Glutamyl hydrolase on methotrexate metabolism and resistance. AB - Intracellular metabolism of methotrexate (MTX) to MTX-polyglutamates (MTXPG) is one determinant of cytotoxicity. Steady-state accumulation of MTXPG seems to depend on the activity of two enzymes: folylpolyglutamate synthetase (FPGS), which adds glutamate residues, and gamma-glutamyl hydrolase (GGH), which removes them. Overexpression of GGH would be expected to decrease intracellular MTXPG, thereby increasing efflux of MTX and decreasing cytotoxicity. Increased expression of GGH has been shown to be associated with resistance to MTX in human sarcoma cell lines and a rat hepatoma cell line. To clarify the specific role of GGH in determining MTX sensitivity, we investigated the phenotype produced by forced GGH overexpression in two cell types. Furthermore, because MTX and folic acid share metabolic pathways, we measured the effects of GGH overexpression on folic acid metabolism. The full-length cDNA for GGH, subcloned into a constitutive expression vector, was transfected into a human fibrosarcoma (HT 1080) and a human breast carcinoma (MCF-7) cell line. Compared with the clones containing an empty vector, the GGH-overexpressing cells express 15- to 30-fold more GGH mRNA, more GGH protein, and 15- to 90-fold more GGH enzyme activity. GGH overexpression altered MTX accumulation and metabolism to long-chain polyglutamates. In contrast to expectations, however, GGH overexpression did not confer resistance to short MTX exposures in either cell line. Changes in MTX metabolism were found to be balanced by alterations in accumulation and metabolism of folic acid. The ratio of MTX:folate accumulation may be a better predictor of MTX cytotoxicity than the accumulation of either alone. We conclude that, at least for these two cell lines, GGH overexpression alone is insufficient to produce clinical resistance to MTX. PMID- 11389097 TI - Overexpression of sialyltransferase CMP-sialic acid:Galbeta1,3GalNAc-R alpha6 Sialyltransferase is related to poor patient survival in human colorectal carcinomas. AB - Thomsen-Friedenreich (TF)-related blood group antigens, such as TF, Tn, and their sialylated variants, belong to a family of tumor-associated carbohydrates. The aim of the present study was to examine tumor-associated alterations of glycosyltransferases involved in the biosynthesis of the TF glycotope in colorectal carcinomas. To this end, glycosyltransferase expression was examined in 40 cases of colorectal carcinoma specimens classified according to the WHO/Union International Contre Cancer guidelines and in "normal" mucosa of the same patients. Occurrence of TF glycotope was examined by immunohistochemistry with the monoclonal antibody A78-G/A7. Expression of sialyltransferases CMP sialic acid:Galbeta1,3GalNAc-R alpha3-sialyltransferase I and II (ST3Gal-I and ST3Gal-II) and CMP-sialic acid:Galbeta1,3GalNAc-R alpha6-sialyltransferase (ST6GalNAc-II) and of core 2 beta1,6-N-acetylglucosaminyltransferase was determined by reverse transcription-PCR in the same cryostat sections used for immunohistochemistry. Additionally, alpha2,3-sialyltransferase enzyme activity was studied in each of these tissues. The TF glycotope was detected in 7% of the normal mucosa, but in 57% of the carcinoma samples. Expression of alpha2,3 sialyltransferases ST3Gal-I, ST3Gal-II, and enzyme activity of alpha2,3 sialyltransferase was significantly increased (P < 0.001) in carcinoma specimens compared with normal mucosa. ST3Gal-I mRNA expression was significantly increased (P = 0.05) in cases showing invasion of lymph vessels. Expression of ST6GalNAc-II was significantly increased (P = 0.04) in cases with metastases to lymph nodes along the vascular trunk. Moreover, ST6GalNAc-II expression provides an prognostic factor for patient survival (log rank, P = 0.02). In an attempt to study the functional relevance of the glycosyltransferases for TF biosynthesis, SW480 colorectal cells were transfected with each of the enzymes, and cell surface expression of the TF glycotope was examined by flow cytometry. The presence of TF was not altered by transfection of the cells with either sialyltransferase ST3Gal-I or ST3Gal-II. However, successful transfection with core 2 beta1,6-N-acetylglucosaminyltransferase led to reduced expression of TF. In contrast, increased cell surface expression of TF was found after ST6GalNAc-II transfection. Thus, expression of TF on the cell surface of SW480 colorectal carcinoma cells depends on the ratio of core 2 beta1,6-N acetylglucosaminyltransferase and ST6GalNAc-II. Earlier immunohistological studies demonstrated that TF is a prognostic factor for patient survival. Our results suggest that sialyltransferase ST6GalNAc-II is of crucial relevance for the prognostic significance of TF. PMID- 11389098 TI - Protein kinase Calpha and protein kinase Cdelta play opposite roles in the proliferation and apoptosis of glioma cells. AB - Protein kinase C (PKC) has been implicated in the proliferation and apoptosis of glial tumors, but the role of specific PKC isoforms remains unresolved. Comparing brain tumors differing in degree of malignancy, we found that malignant gliomas expressed higher levels of PKCalpha and lower levels of PKCdelta as compared with low-grade astrocytomas. Consistent with a mechanistic role for these differences, overexpression of PKCalpha in the human U87 glioma cell line resulted in enhanced cell proliferation and decreased glial fibrillary acidic protein (GFAP) expression as compared with controls. Reciprocally, overexpression of PKCdelta inhibited cell proliferation and enhanced GFAP expression. Using PKC chimeras, we found that the regulatory domains of PKCalpha and PKCdelta mediated their effects on cell proliferation and GFAP expression. PKCalpha and delta have been implicated as potential signaling molecules in apoptosis. Therefore, we examined the role of these isoforms in the resistance of glioma cells to apoptotic stimuli. In U87 cells, manipulation of PKCalpha levels had little effect on apoptosis in response to etoposide. In contrast, overexpression of PKCdelta rendered the U87 cells more sensitive to the apoptotic effect of etoposide, and PKCdelta was cleaved in these cells by a caspase-dependent process. Furthermore, the glioma cell line U373, which expresses endogenous PKCdelta, underwent apoptosis in response to etoposide, and the apoptotic response was blocked by the PKCdelta inhibitor rottlerin. Our results suggest that PKCalpha and PKCdelta play opposite roles in the proliferation and apoptosis of glioma cells. PMID- 11389099 TI - Increased expression of UDP-galactose transporter messenger RNA in human colon cancer tissues and its implication in synthesis of Thomsen-Friedenreich antigen and sialyl Lewis A/X determinants. AB - A series of human nucleotide sugar transporters of the Golgi apparatus was recently cloned, including the transporters for UDP-galactose (UDP-Gal), UDP-N acetylglucosamine (UDP-GlcNAc) and CMP-sialic acid (CMP-SA). We have examined the mRNA expression of these three transporters in human colon cancer tissues by reverse transcription-PCR analysis and compared it with that in nonmalignant colonic mucosa prepared from the same patients. The amount of mRNA for UDP-Gal transporter was significantly increased in colon cancer tissues compared with nonmalignant mucosa tissues (P = 0.035; n = 20). The increase was more prominent in patients with advanced colorectal cancer of Dukes' stages C and D, in which the amount of UDP-Gal transporter mRNA in cancer tissues showed on average about a 3.6-fold increase over the paired nonmalignant mucosa (statistically significant at P = 0.004; n = 14). The mRNA content of the other two transporters showed no significant difference between the paired cancer and normal tissues. When UDP-Gal transporter cDNA was stably transfected to cultured human colon cancer cells, the expression of Thomsen-Friedenreich (TF) antigen and of sialyl Lewis A (NeuAcalpha2-->3Galbeta1-->3[Fucalpha1-->4]GlcNAcbeta1-->R) and sialyl Lewis X (NeuAcalpha2-->3Galbeta1-->4[Fucalpha1-->3]GlcNAcbeta1-->R) determinants was significantly induced on transfectant cells, which resulted in markedly enhanced cell adhesion to vascular E-selectin. These findings suggest that the increase of UDP-Gal transporter mRNA is involved in the enhanced expression of cancer-associated carbohydrate determinants such as TF and sialyl Lewis A/X antigens in colon cancers. PMID- 11389100 TI - Transcriptional silencing of Cyclooxygenase-2 by hyper-methylation of the 5' CpG island in human gastric carcinoma cells. AB - It has been well established that overexpression of Cyclooxygenase-2 (Cox-2) in epithelial cells inhibits apoptosis and increases the invasiveness of malignant cells, favoring tumorigenesis and metastasis. However, the molecular mechanism that regulates Cox-2 expression has not been well defined in gastric carcinoma. In this study, we examined whether the Cox-2 expression could be regulated by hyper-methylation of the Cox-2 CpG island (spanning from -590 to +186 with respect to the transcription initiation site) in human gastric carcinoma cell lines. By Southern analysis, we found that three gastric cells (SNU-601, -620, and -719) without Cox-2 expression demonstrated hyper-methylation at the Cox-2 CpG island. A detailed methylation pattern using bisulfite sequencing analysis revealed that all of the CpG sites were completely methylated in SNU-601. Treatment with demethylating agents effectively reactivated the expression of Cox 2 and restored IL-1beta sensitivity in the previously resistant SNU-601. By transient transfection experiments, we demonstrate that constitutively active Cox 2 promoter activities were exhibited even without an exogenous stimulation in SNU 601. Furthermore, when the motif of the nuclear factor for interleukin-6 expression site, the cyclic AMP response element, or both was subjected to point mutation, the constitutive luciferase activity was markedly reduced. In addition, Cox-2 promoter activity was completely blocked by in vitro methylation of all of the CpG sites in the Cox-2 promoter region with SssI (CpG) methylase in SNU-601. Taken together, these results indicate that transcriptional repression of Cox-2 is caused by hyper-methylation of the Cox-2 CpG island in gastric carcinoma cell lines. PMID- 11389101 TI - Y(1)-mediated effect of neuropeptide Y in cancer: breast carcinomas as targets. AB - Overexpression of selected peptide receptors in human tumors has been shown to represent clinically relevant targets for cancer diagnosis and therapy. Neuropeptide Y (NPY) is a peptide neurotransmitter mediating feeding behavior and vasoconstriction. It has never been shown to be involved in human cancer. We show here, using in vitro receptor autoradiography, a NPY receptor incidence of 85% in primary human breast carcinomas (n = 95) and of 100% in lymph node metastases of receptor-positive primaries (n = 27), predominantly as Y(1) subtype, whereas non neoplastic human breast expressed Y(2) preferentially. Y(1) mRNA was detected in Y(1)-expressing tumors by in situ hybridization, whereas Y(2) mRNA was found in normal breast tissue. The strong predominance of Y(1) in breast carcinomas compared with Y(2) in normal breast suggests that neoplastic transformation can switch the NPY receptor expression from Y(2) to Y(1) subtype. Moreover, in Y(1) expressing human SK-N-MC tumor cells, an NPY-induced, dose-dependent inhibition of tumor cell growth of >40% was observed, suggesting a functional role of NPY in cancer, mediated by Y(1). NPY should therefore be added to the list of small regulatory peptides related to cancer. The high incidence of Y(1) in in situ, invasive, and metastatic breast cancers allows for the possibility to target them for diagnosis and therapy with NPY analogues. PMID- 11389102 TI - Melanoma biology and progression. PMID- 11389105 TI - Characterization of class 1 integrons associated with R-plasmids in clinical Aeromonas salmonicida isolates from various geographical areas. AB - Class 1 integrons were found in 26 of 40 antibiotic-resistant isolates of the fish pathogen Aeromonas salmonicida from Northern Europe and North America. Three different dhfr genes, conferring trimethoprim resistance, and one ant(3")1a aminoglycoside resistance gene were identified as gene inserts. The gene cassettes tended to be conserved among isolates from a particular geographical area. Nineteen isolates transferred R-plasmids carrying different tet determinants to Escherichia coli in filter mating assays, and in 15 cases, the class 1 integrons were co-transferred. Transferable sulphadiazine, trimethoprim and streptomycin resistances were invariably encoded by integrons. It thus appears that integron-encoded antibiotic resistance genes contribute substantially to the horizontal spread of antimicrobial resistance within this species, being associated with conjugative plasmids. PMID- 11389106 TI - Characterization, cloning and sequence analysis of the inducible Ochrobactrum anthropi AmpC beta-lactamase. AB - Ochrobactrum anthropi is resistant to most cephalosporins and penicillins due, at least in part, to the inducible expression of a single beta-lactamase. The beta lactamase gene has been cloned and sequenced. It encodes an AmpC-type class 1 serine active-site enzyme that hydrolyses mainly cephalosporins and is resistant to inhibition by clavulanic acid. Expression of the ampC gene is inducible via a typical AmpR regulator, which is encoded upstream of ampC. Inducible expression is retained following cloning of O. anthropi ampR-ampC into Escherichia coli, confirming that the signal for AmpR activation in O. anthropi is the same as that used in the Enterobacteriaceae. This is the first reported example of an AmpC beta-lactamase outside of the gamma-subdivision of the bacterial kingdom. Genomic searches of other non-gamma-subdivision bacteria revealed a homologous ampR-ampC cluster in the plant symbiont, Sinorhizobium meliloti. PMID- 11389107 TI - Outer membrane protein change combined with co-existing TEM-1 and SHV-1 beta lactamases lead to false identification of ESBL-producing Klebsiella pneumoniae. AB - Nine isolates of Klebsiella pneumoniae, obtained from one colonized and eight bacteraemic patients on a paediatric ward, were shown to be identical by PFGE, indicating an outbreak. Screening for extended-spectrum beta-lactamase (ESBL) production using the double-disc synergy test, Etest for ESBLs and agar diffusion tests indicated ESBL production. The isolates showed reduced susceptibility to cefotaxime but not to other third-generation cephalosporins. Molecular studies revealed production of TEM-1 and SHV-1 but no ESBLs were identified. Deficiency in expression of an outer membrane protein (OmpK35) was also observed. These observations led us to postulate that the extremely low level of OmpK35 expression and the co-existence of TEM-1 and SHV-1 resulted in an increased MIC of cefotaxime and the false designation of the isolates as ESBL producers. All the infected infants were treated with either third-generation cephalosporins alone or multiple antibiotics including a third-generation cephalosporin, and recovered and were discharged without sequelae. PMID- 11389108 TI - Antibiotic resistance of faecal Escherichia coli in poultry, poultry farmers and poultry slaughterers. AB - The percentage of faecal samples containing resistant Echerichia coli and the proportion of resistant faecal E. coli were determined in three poultry populations: broilers and turkeys commonly given antibiotics, and laying hens treated with antibiotics relatively infrequently. Faecal samples of five human populations were also examined: turkey farmers, broiler farmers, laying-hen farmers, broiler slaughterers and turkey slaughterers. The MICs of antibiotics commonly used in poultry medicine were also determined. Ciprofloxacin-resistant isolates from these eight populations and from turkey meat were genotyped by pulsed-field gel electrophoresis (PFGE) after SmaI digestion. The proportion of samples containing resistant E. coli and the percentages of resistant E. coli were significantly higher in turkeys and broilers than in the laying-hen population. Resistance to nearly all antibiotics in faecal E. coli of turkey and broiler farmers, and of turkey and broiler slaughterers, was higher than in laying-hen farmers. Multiresistant isolates were common in turkey and broiler farmers but absent in laying-hen farmers. The same resistance patterns were found in turkeys, turkey farmers and turkey slaughterers and in broiler, broiler farmers and broiler slaughterers. The PFGE patterns of the isolates from the eight populations were quite heterogeneous, but E. coli with an identical PFGE pattern were isolated at two farms from a turkey and the farmer, and also from a broiler and a broiler farmer from different farms. Moreover, three E. coli isolates from turkey meat were identical to faecal isolates from turkeys. The results of this study strongly indicate that transmission of resistant clones and resistance plasmids of E. coli from poultry to humans commonly occurs. PMID- 11389109 TI - Antibiotic susceptibility of bacterial strains isolated from urinary tract infections in Poland. AB - Worldwide data show that there is increasing resistance among urinary tract pathogens to conventional drugs. The aim of this study was to obtain data on susceptibility patterns of pathogens responsible for urinary tract infections (UTIs) in Poland to currently used antimicrobial agents. A multicentre study of 141 pathogens from hospital-acquired infections and 460 pathogens from community acquired infections was carried out between July 1998 and May 1999. The most prevalent aetiological agent was Escherichia coli (73.0%), followed by Proteus spp. (8.9%) and other species of Enterobacteriaceae (9.6%). Few community infections were caused by Gram-positive bacteria (2.2%). Gram-positive cocci were isolated more frequently from a hospital setting (14.1%) and the most common were Enterococcus spp. (8.5%). Pseudomonas aeruginosa was found only among hospital isolates and was responsible for 10.7% of infections. E. coli isolates from both community and hospital infections were highly susceptible to many antimicrobial agents with the exception of those isolates producing extended-spectrum beta lactamases (ESBLs). Of all Enterobacteriaceae tested, 38 strains (6.9%) were capable of producing ESBLs. PMID- 11389110 TI - Prior trimethoprim use and trimethoprim-resistant urinary tract infection: a nested case-control study with multivariate analysis for other risk factors. AB - Trimethoprim resistance is increasingly prevalent in community-acquired urinary infections. The objective of this study was to evaluate the association between exposure to community-prescribed trimethoprim and other risk factors in subjects and subsequent trimethoprim-resistant urinary tract infection. The design was a nested case-control study using a record-linkage database. Study subjects submitted a urine sample to the Ninewells Hospital Laboratory between July 1993 and December 1995. Antibiotic exposure in subjects with trimethoprim-resistant isolates (cases) was compared with antibiotic exposure in subjects with trimethoprim-susceptible isolates (controls). Study subjects were drawn from the catchment area of a large teaching hospital in Tayside, Scotland. There were 13765 males and females aged 1-106 years who submitted their first urine sample for culture during the study period. After adjustment for significant risk factors and confounding variables, logistic regression analysis showed exposure to trimethoprim [odds ratio (OR) 4.35] or any antibiotic other than trimethoprim (OR 1.32) to be predictive of resistance. The growth of Proteus spp. (OR 115.14) and bacterial growth other than Escherichia coli and Proteus spp. (OR 2.83) were also predictor variables. Hospitalization in the previous 6 months was not independently associated with trimethoprim resistance. In conclusion, trimethoprim resistance was independently associated with exposure to trimethoprim and to antibiotics other than trimethoprim. Reduction in trimethoprim prescribing alone may not reduce the prevalence of trimethoprim resistance. PMID- 11389111 TI - Antimicrobial susceptibility of Pseudomonas aeruginosa: results of a UK survey and evaluation of the British Society for Antimicrobial Chemotherapy disc susceptibility test. AB - A survey was conducted in 1999, first to establish the prevalence of antibiotic resistance among clinical isolates of Pseudomonas aeruginosa in the UK and secondly to test whether the use of the standardized British Society for Antimicrobial Chemotherapy (BSAC) disc testing method improved the accuracy of routine susceptibility testing for this organism. Twenty-five hospitals were each asked to collect up to 100 consecutive, clinically significant isolates of P. aeruginosa and to test their susceptibility to amikacin, gentamicin, ceftazidime, imipenem, meropenem, ciprofloxacin, piperacillin and piperacillin/tazobactam using the new BSAC disc method. A total of 2194 isolate reports were available for analysis and 10% of the isolates represented, plus those with unusual resistances, were re-tested centrally for quality control purposes. The zone distributions were essentially unimodal, indicating the absence of major populations with acquired resistance. The results indicated that resistance rates to the beta-lactam, aminoglycoside and quinolone agents tested in P. aeruginosa in the UK remain low (<12%), and were mostly unchanged since a previous survey conducted in 1993. High resistance rates were nevertheless reported for isolates from cystic fibrosis patients. The accuracy of susceptibility testing using the new BSAC disc testing method was better than in previous studies, when Stokes' method was most frequently used. Critically, the proportion of resistant isolates incorrectly reported as susceptible was reduced significantly; nevertheless, depending on the antibiotic, up to 49% of the isolates reported as intermediate or resistant were found susceptible on central re-testing. PMID- 11389112 TI - External quality assessment of antimicrobial susceptibility testing in Europe. AB - Comparability of results of antimicrobial susceptibility testing is essential for resistance surveillance studies. As different methods may be used in different countries, there may be particular problems with international comparisons of resistance rates. Data from external quality assessment (EQA) surveys participated in by laboratories from several European countries allow comparison of performance between countries. In this study, success rates with organism antimicrobial agent combinations known to be difficult to test were examined. With penicillin resistance in pneumococci; vancomycin and high-level gentamicin resistance in enterococci; ampicillin, co-amoxiclav and chloramphenicol resistance in Haemophilus influenzae and methicillin resistance in staphylococci there were differences between countries in success rates for discrimination of resistant strains. This study suggests that differences between countries in rates of resistance for some organism-antimicrobial agent combinations should be interpreted with caution. International EQA is useful in the demonstration and clarification of such differences. PMID- 11389113 TI - Pharmacodynamics of moxifloxacin, levofloxacin and sparfloxacin against Streptococcus pneumoniae. AB - An in vitro pharmacokinetic model (IVPM) was used to simulate the human serum pharmacokinetics of moxifloxacin, levofloxacin and sparfloxacin, and to compare their pharmacodynamics against Streptococcus pneumoniae exhibiting a wide range of susceptibilities to fluoroquinolones. Logarithmic-phase cultures were exposed to peak concentrations achieved in human serum of moxifloxacin, levofloxacin or sparfloxacin with oral doses of 400, 500 and 200 mg, respectively. Human elimination pharmacokinetics were simulated, and viable bacterial counts were measured at 0, 1, 2, 4, 6, 8, 24 and 36 h. Moxifloxacin was rapidly bactericidal (>3 logs of killing) against all 10 S. pneumoniae strains, with 99.9% kills of eight strains occurring within 1-3 h after dosing. Maximum kills ranged from 5 to >6 logs. Moxifloxacin eradicated seven strains from the IVPM within 8 h of the first dose, and eradicated two other strains within 24 h. Although levofloxacin and sparfloxacin were also bactericidal against all 10 S. pneumoniae strains, the rates of killing were somewhat slower, with sparfloxacin exhibiting the slowest rate of kill. In summary, moxifloxacin's increased anti-pneumococcal potency compared with levofloxacin and its more favourable pharmacokinetics compared with sparfloxacin provided enhanced pharmacodynamic activity against some S. pneumoniae strains when maximum doses were simulated in an IVPM. PMID- 11389114 TI - Erythrocyte-mediated delivery of a new homodinucleotide active against human immunodeficiency virus and herpes simplex virus. AB - Monocyte-derived macrophages (MDMs) play a central role in the pathogenesis of infection by human immunodeficiency virus (HIV-1) and represent one of the main reservoirs of the virus in the body. In addition, MDMs can easily be infected by various herpes viruses, including herpes simplex virus type 1 (HSV-1). We have synthesized a new antiviral agent (Bis-PMEA) that consists of two 9-(2 phosphonylmethoxyethyl)adenine (PMEA) molecules bound by a phosphate bridge. This nucleotide analogue, like the parent compound PMEA, has strong and selective activity against HIV-1 and HSV-1. A drug-targeting system previously developed in our laboratory was used for the selective delivery of these drugs to macrophages. Bis-PMEA and PMEA were encapsulated into autologous erythrocytes by a procedure of hypotonic dialysis and isotonic resealing. Loaded erythrocytes were modified to increase their recognition and phagocytosis by human macrophages. By administering Bis-PMEA-loaded erythrocytes to macrophages, 47% of Bis-PMEA and 28% of PMEA was still present 10 days after phagocytosis; in contrast, only 12% of PMEA was found in macrophages receiving PMEA-loaded erythrocytes. Bis-PMEA loaded erythrocytes were then added to macrophages infected with HIV-1 and HSV-1 and their antiviral activity evaluated. Remarkable protection was obtained against HIV-1 and HSV-1 infection (95 and 85%, respectively). Therefore, Bis-PMEA acts as an efficient antiviral prodrug that, following selective targeting to macrophages by means of loaded erythrocytes, can protect a refractory cell compartment. PMID- 11389116 TI - Lung concentrations of telithromycin after oral dosing. AB - Concentrations of telithromycin were measured in plasma, bronchial mucosa (BM), epithelial lining fluid (ELF) and alveolar macrophages (AM) following multiple oral doses. Concentrations were determined using a microbiological assay. There were 20 subjects in the study, allocated to three nominal time periods: 2, 12 and 24 h. Mean concentrations in plasma, BM, ELF and AM for 2, 12 and 24 h were as follows: 2 h, 1.86 mg/L, 3.88 mg/kg, 14.89 mg/L and 69.32 mg/L; 12 h, 0.23 mg/L, 1.41 mg/kg, 3.27 mg/L and 318.1 mg/L; and 24 h, 0.08 mg/L, 0.78 mg/kg, 0.97 mg/L and 161.57 mg/L. These concentrations of telithromycin in BM and ELF exceeded for 24 h the mean MIC90s of the common respiratory pathogens Streptococcus pneumoniae (0.12 mg/L) and Moraxella catarrhalis (0.03 mg/L), as well as the atypical microorganism Mycoplasma pneumoniae (0.001 mg/L), and suggest that telithromycin may be effective for the treatment of community-acquired pneumonia and chronic obstructive pulmonary disease. PMID- 11389115 TI - Chemotherapy of Mycobacterium tuberculosis infections in mice with a combination of isoniazid and rifampicin entrapped in Poly (DL-lactide-co-glycolide) microparticles. AB - Strategies to improve patient compliance in tuberculosis chemotherapy include the use of sustained release drug delivery systems. In this study, Poly (DL-lactide co-glycolide) (PLG) microparticles containing a combination of isoniazid and rifampicin were developed as sustained release carrier systems. A single dose of PLG microparticles exhibited a sustained release of isoniazid and rifampicin in vivo up to 7 and 6 weeks, respectively. Free drugs (in combination) injected in the same doses were detectable in vivo up to 24 h only. One dose of PLG microparticles cleared bacteria more effectively from lungs and liver in an experimental murine model of tuberculosis after low-dose PLG combination drug therapy and in liver after high-dose PLG combination drug therapy as compared with a daily administration of the free drugs. These results suggest that PLG microparticles offer an improvement for tuberculosis chemotherapy over the conventional treatment. PMID- 11389117 TI - Meropenem versus ceftazidime as empirical monotherapy in febrile neutropenia of paediatric patients with cancer. AB - This trial assessed the efficacy and safety of meropenem versus ceftazidime as empirical monotherapy for febrile neutropenia in paediatric cancer patients. In a prospective randomized study, 172 evaluable febrile episodes in the meropenem arm and 170 episodes in the ceftazidime arm were analysed for the clinical and microbiological response dependent on the kind of infection. About half the episodes were classified as fever of unknown origin (FUO) and the remainder as microbiologically or clinically documented infections. The most frequently documented infections in both arms were bacteraemias (22.1 versus 26.5%), predominantly caused by Gram-positive organisms (57.9 versus 71.1%). The success rate of the initial monotherapy differed significantly between the two arms and was 55.8% in the meropenem and 40.0% in the ceftazidime arm (P = 0.003). In addition, a significantly longer duration of fever and of antimicrobial therapy was observed in the ceftazidime arm than in the meropenem arm (median 5 versus 4 days, P = 0.022, and 7 versus 6 days, P = 0.009, respectively). With respect to the kind of infection, differences between the two arms were significant only in episodes classified as FUO but not in documented infections. In both arms, side effects were minimal. Despite the greater response rate for meropenem in FUO, the fact that ceftazidime has been proven to be as effective as meropenem in documented infections in the present study suggests that both drugs are useful as empirical monotherapy in febrile paediatric cancer patients. PMID- 11389118 TI - Comparable aciclovir exposures produced by oral valaciclovir and intravenous aciclovir in immunocompromised cancer patients. AB - The objective of this study was to evaluate the comparability of systemic aciclovir exposure at steady state in immunocompromised patients following oral valaciclovir 1000 mg tds and intravenous (iv) aciclovir 5 mg/kg tds. A two centre, randomized, open label, two-way crossover study was undertaken. Patients aged 18-65 years who had undergone high-dose chemotherapy for cancer and were neutropenic (neutrophil count <0.5 x 109/mL) with normal renal function were recruited. The pharmacokinetic parameters of aciclovir after oral valaciclovir 1000 mg or iv aciclovir 5 mg/kg given as 1 h infusion, each administered every 8 h for seven doses, were compared. Fifteen patients were enrolled and 13 completed both treatments. The mean (s.d.) values for aciclovir after oral valaciclovir and iv aciclovir were: AUC0-8 76.3 (29.7) and 64.2 (20.0) microM x h; peak plasma concentration (Cmax) 26.6 (10.5) and 34.0 (11.9) microM; time to maximal plasma concentration (tmax) 2.01 (0.65) and 0.95 (0.19); and plasma elimination half life (t1/2) 2.83 (0.91) and 2.44 (0.62) h, respectively. The mean absolute bioavailability of aciclovir from oral valaciclovir was 60 +/- 21%. Equivalent systemic exposure to aciclovir after oral valaciclovir 1000 mg and iv aciclovir 5 mg/kg was observed with AUC0-8 (oral/iv ratio = 1.16; 90% CI 0.98-1.39), whilst significantly reduced peak aciclovir concentrations were obtained with oral valaciclovir (ratio = 0.75; 90% CI 0.60-0.94). Oral valaciclovir offers a convenient, and possibly safer, alternative to iv aciclovir, delivering comparable systemic exposures with reduced peak levels. This may contribute to shorter hospitalization, reduced costs for healthcare providers and improved quality of life for patients. PMID- 11389119 TI - Distribution of resistance genes tet(M), aph3'-III, catpC194 and the integrase gene of Tn1545 in clinical Streptococcus pneumoniae harbouring erm(B) and mef(A) genes in Spain. AB - The most prevalent macrolide resistance phenotype and genotype among pneumococcal isolates was the cMLSB phenotype [erm(B) or erm(B)/mef(A)] (91.3%). We studied the distribution of other resistance genes, tet(M), catpC194, aph3'-III, in these strains, seeing evolution at work in that some strains carried different combinations of resistance determinants. The most prevalent patterns associated with resistance to erythromycin [erm(B)] were resistance to tetracycline [tet(M)] and chloramphenicol (catpC194) (48.2%) or resistance to tetracycline [tet(M)] alone (42.2%). In our isolates of Streptococcus pneumoniae there was a strong association of the erm(B) and tet(M) genes with Tn1545-related elements. PMID- 11389120 TI - The effect of azithromycin and clarithromycin on ex vivo interleukin-8 (IL-8) release from whole blood and IL-8 production by human alveolar macrophages. AB - We investigated the effects of azithromycin and clarithromycin, two antibiotics that possess a broad spectrum of antimicrobial activity (including antimycobacterial activity), on interleukin-8 (IL-8) release from human whole blood leucocytes and lung macrophages. Ex vivo stimulation of leukocytes with either of the antibiotics (0.04-40 mg/L) significantly increased IL-8 secretion. Incubation of alveolar macrophages with different concentrations of azithromycin or clarithromycin modified IL-8 production: it increased at a drug concentration of 4 mg/L and decreased at concentration of 400 mg/L. Our findings suggest that azithromycin and clarithromycin may alter IL-8 production, thus enhancing the clinical effectiveness of these antibiotics. PMID- 11389121 TI - Enhanced fungicidal activity of N-chlorotaurine in nasal secretion. AB - The antifungal activity of N-chlorotaurine (NCT), a long-lived oxidant produced by stimulated human leucocytes, was investigated. Incubation of Aspergillus spp., Candida spp., Fusarium spp., Penicillium spp. and Alternaria spp. in 1% NCT (55 mM) for 1-4 h produced a log10 reduction in cfu of between 1 and 4. In samples of nasal secretion, killing was significantly hastened (30 min), which may be explained by the formation of monochloramine by halogenation of ammonium, which was found at a concentration of 1 mM in these samples. For these reasons, NCT is of interest as a new agent for treatment of local inflammatory mycosis, e.g. eosinophilic fungal rhinosinusitis. PMID- 11389122 TI - Antimicrobial activity of moxifloxacin, gatifloxacin and six fluoroquinolones against Streptococcus pneumoniae. AB - The in vitro and pharmacodynamic effects of moxifloxacin and gatifloxacin against Streptococcus pneumoniae were compared with six other fluoroquinolones. Organisms included penicillin-susceptible (54) and penicillin-non-susceptible (145) isolates from 1998-1999. Moxifloxacin and clinafloxacin demonstrated the greatest in vitro activity, with MIC90s of 0.13 mg/L, followed by trovafloxacin, grepafloxacin > gatifloxacin, sparfloxacin > levofloxacin > ciprofloxacin. There was no difference in fluoroquinolone activity between penicillin-susceptible and non-susceptible strains. Pharmacodynamic analysis using published pharmacokinetic information indicates that all the agents tested except ciprofloxacin had an AUC/MIC90 > 30, with moxifloxacin having the greatest free-drug availability. PMID- 11389123 TI - Post-antibiotic growth suppression of linezolid against Gram-positive bacteria. AB - The in vitro post-antibiotic effects (PAEs) of eight different concentrations of linezolid against Gram-positive cocci were investigated and the results analysed using the sigmoid E(max) model for mathematically modelling the PAE. Mean maximal linezolid PAEs against strains of Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium and Streptococcus pneumoniae were 2.2, 1.8, 2.8, 2.0 and 3.0 h, respectively. Resistance to methicillin (for the staphylococci), vancomycin (for the enterococci) and penicillin (for the pneumococci) had no effect on the duration of the PAE. Results of PAE testing support twice-daily dosing of linezolid in humans. PMID- 11389124 TI - Biomaterial-associated infection of gentamicin-loaded PMMA beads in orthopaedic revision surgery. AB - In two-stage orthopaedic revision surgery, high local levels of antibiotics are achieved after removal of an infected prosthesis through temporary implantation of gentamicin-loaded beads. However, despite their antibiotic release, these beads act as a biomaterial surface to which bacteria preferentially adhere, grow and potentially develop antibiotic resistance. Gentamicin-loaded beads were retrieved from 20 patients with prosthesis-related infections. Excised tissue samples were taken for routine culture, while beads were analysed in an extensive laboratory procedure. Extensive culture procedures indicated the presence of bacteria on gentamicin-loaded beads in 18 of the 20 patients involved, while 12 of these 18 patients were considered free of infection by routine culture. Nineteen of 28 bacterial strains isolated were gentamicin resistant and cultures from three patients yielded highly gentamicin-resistant sub-populations. It is concluded that routine culture of excised tissues in orthopaedic revision surgery is inadequate to ascertain full eradication of infection, especially as infecting, antibiotic-resistant bacteria preferentially adhere to and grow on gentamicin-loaded beads. Extensive examination of the bead surfaces is proposed as a more reliable indication that infection has been eradicated. PMID- 11389125 TI - Quinolone-induced QT interval prolongation: a not-so-unexpected class effect. PMID- 11389126 TI - Conventional dogma applied to quinolones? Time for a change. PMID- 11389127 TI - No evidence that rifampicin has glucocorticoid-like immunosuppressive properties leading to suppression of rat-splenocyte proliferation in vitro. PMID- 11389128 TI - Distribution of the antiseptic resistance genes qacA, qacB and qacC in 497 methicillin-resistant and -susceptible European isolates of Staphylococcus aureus. PMID- 11389129 TI - Accuracy of routine susceptibility testing of Streptococcus pneumoniae in England and Wales. PMID- 11389130 TI - In vitro antimicrobial activities of a novel everninomicin for multiple drug resistant Streptococcus pneumoniae isolates in Japan. PMID- 11389131 TI - Study of vancomycin tolerance in 120 strains of Streptococcus pneumoniae isolated in 1999 in Madrid, Spain. PMID- 11389132 TI - Comparative study of antimicrobial resistance of Escherichia coli strains isolated from urinary tract infection in patients from Caracas and Lima. PMID- 11389133 TI - In vitro activity of rifaximin against bacterial enteropathogens causing diarrhoea in children under 5 years of age in Ifakara, Tanzania. PMID- 11389134 TI - Squamous cell tumors in mice heterozygous for a null allele of Atp2a2, encoding the sarco(endo)plasmic reticulum Ca2+-ATPase isoform 2 Ca2+ pump. AB - Mutations in the human ATP2A2 gene, encoding sarco(endo)plasmic reticulum Ca(2+) ATPase isoform 2 (SERCA2), cause Darier disease, an autosomal dominant skin disease characterized by multiple keratotic papules in the seborrheic regions of the body. Mice with a single functional Atp2a2 allele (the mouse homolog of ATP2A2) were shown previously to have reduced levels of SERCA2 in heart and mildly impaired cardiac contractility and relaxation. Here we show that aged heterozygous mutant (Atp2a2(+/-)) mice develop squamous cell tumors of the forestomach, esophagus, oral mucosa, tongue, and skin. Squamous cell tumors occurred in 13/14 Atp2a2(+/-) mice but were not observed in age- and sex-matched wild-type controls. Hyperkeratinized squamous cell papillomas and carcinomas of the upper digestive tract were the most frequent finding among Atp2a2(+/-) mice, and many animals had multiple tumors. Western blot analyses showed that SERCA2 protein levels were reduced in skin and other affected tissues of heterozygous mice. The development of squamous cell tumors in aged Atp2a2(+/-) mice indicates that SERCA2 haploinsufficiency predisposes murine keratinocytes to neoplasia. These findings provide the first direct demonstration that a perturbation of Ca(2+) homeostasis or signaling can serve as a primary initiating event in cancer. PMID- 11389135 TI - Heme histidine ligands within gp91(phox) modulate proton conduction by the phagocyte NADPH oxidase. AB - The membrane subunit of the phagocyte NADPH oxidase, gp91(phox), possesses a H(+) channel motif formed by membrane-spanning histidines postulated to coordinate the two heme groups forming the redox center of the flavocytochrome. To study the role of heme-binding histidines on proton conduction, we stably expressed the gp91(phox) cytochrome in human embryonic kidney 293 cells and measured proton currents with the patch clamp technique. Similar to its shorter homologue, NADPH oxidase homologue 1, which is predicted not to bind heme, gp91(phox) generated voltage-activated, pH-dependent, H(+)-selective currents that were reversibly blocked by Zn(2+). The gp91(phox) currents, however, activated faster, deactivated more slowly, and were markedly affected by the inhibition of heme synthesis. Upon heme removal, the currents had larger amplitude, activated faster and at lower voltages, and became sensitive to the histidine reagent diethylpyrocarbonate. Mutation of the His-115 residue to leucine abolished both the gp91(phox) characteristic 558-nm absorbance peak and voltage-activated currents, indicating that His-115 is involved in both heme ligation and proton conduction. These results indicate that the gp91(phox) proton channel is activated upon release of heme from its His-115 ligand. During activation of the oxidase complex, changes in heme coordination within the cytochrome might increase the mobility of histidine ligands, thereby coupling electron and proton transport. PMID- 11389137 TI - Clathrin box in G protein-coupled receptor kinase 2. AB - beta(1)-Adrenergic receptor (beta(1)AR) shows the resistance to agonist-induced internalization. However, beta(1)AR can internalize as G protein-coupled receptor kinase 2 (GRK2) is fused to its carboxyl terminus. Internalization of the beta(1)AR and GRK2 fusion protein (beta(1)AR/GRK2) is dependent on dynamin but independent of beta-arrestin and phosphorylation. The beta(1)AR/GRK2 fusion protein internalizes via clathrin-coated pits and is found to co-localize with the endosome that contains transferrin. The fusion proteins consisting of beta(1)AR and various portions of GRK2 reveal that the residues 498-502 in the carboxyl-terminal domain of GRK2 are critical to promote internalization of the fusion proteins. This domain contains a consensus sequence of a clathrin-binding motif defined as a clathrin box. In vitro binding assays show that the residues 498-502 of GRK2 bind the amino-terminal domain of clathrin heavy chain to almost the same extent as beta-arrestin1. The mutation of the clathrin box in the carboxyl-terminal domain of GRK2 results in the loss of the ability to promote internalization of the fusion protein. GRK2 activity increases and then decreases as the concentration of clathrin heavy chain increases. Taken together, these results imply that GRK2 contains a functional clathrin box and directly interacts with clathrin to modulate its function. PMID- 11389136 TI - Processive phosphorylation of p130Cas by Src depends on SH3-polyproline interactions. AB - Many in vivo substrates of Src family tyrosine kinases possess sequences conforming to Src homology 2 and 3 (SH2 and SH3) domain-binding motifs. One such substrate is p130Cas, a protein that is hyperphosphorylated in v-Src transformed cells. Cas contains a substrate domain consisting of 15 potential tyrosine phosphorylation sites, C- and N-terminal polyproline regions fitting the consensus sequence for SH3 domain ligands, and a YDYV motif that binds the Src SH2 domain when phosphorylated. In an effort to understand the mechanisms of processive phosphorylation, we have explored the regions of Cas necessary for interaction with Src using the yeast two-hybrid system. Mutations in the SH2 domain-binding region of Cas or the Src SH2 domain have little effect in Cas-Src complex formation or phosphorylation. However, disruption of the C-terminal polyproline region of Cas completely abolishes interaction between the two proteins and results in impaired phosphorylation of Cas. Kinetic analyses using purified proteins indicated that multisite phosphorylation of Cas by Src follows a processive rather than a distributive mechanism. Furthermore, the kinetic studies show that there are two properties of the polyproline region of Cas that are important in enhancing substrate phosphorylation. First, the C-terminal polyproline serves to activate Src kinases through the process of SH3 domain displacement. Second, this region aids in anchoring the kinase to Cas to facilitate processive phosphorylation of the substrate domain. The two processes combine to ensure phosphorylation of Cas with high efficiency. PMID- 11389138 TI - Selenium metabolism in Drosophila: selenoproteins, selenoprotein mRNA expression, fertility, and mortality. AB - Selenocysteine is a rare amino acid in protein that is encoded by UGA with the requirement of a downstream mRNA stem-loop structure, the selenocysteine insertion sequence element. To detect selenoproteins in Drosophila, the entire genome was analyzed with a novel program that searches for selenocysteine insertion sequence elements, followed by selenoprotein gene signature analyses. This computational screen and subsequent metabolic labeling with (75)Se and characterization of selenoprotein mRNA expression resulted in identification of three selenoproteins: selenophosphate synthetase 2 and novel G-rich and BthD selenoproteins that had no homology to known proteins. To assess a biological role for these proteins, a simple chemically defined medium that supports growth of adult Drosophila and requires selenium supplementation for optimal survival was devised. Flies survived on this medium supplemented with 10(-8) to 10(-6) m selenium or on the commonly used yeast-based complete medium at about twice the rate as those on a medium without selenium or with >10(-6) m selenium. This effect correlated with changes in selenoprotein mRNA expression. The number of eggs laid by Drosophila was reduced approximately in half in the chemically defined medium compared with the same medium supplemented with selenium. The data provide evidence that dietary selenium deficiency shortens, while supplementation of the diet with selenium normalizes the Drosophila life span by a process that may involve the newly identified selenoproteins. PMID- 11389139 TI - The yeast a-factor transporter Ste6p, a member of the ABC superfamily, couples ATP hydrolysis to pheromone export. AB - ATP-binding cassette (ABC) proteins transport a diverse collection of substrates. It is presumed that these proteins couple ATP hydrolysis to substrate transport, yet ATPase activity has been demonstrated for only a few. To provide direct evidence for such activity in Ste6p, the yeast ABC protein required for the export of a-factor mating pheromone, we established conditions for purification of Ste6p in biochemical quantities from both yeast and Sf9 insect cells. The basal ATPase activity of purified and reconstituted Ste6p (V(max) = 18 nmol/mg/min; K(m) for MgATP = 0.2 mm) compares favorably with several other ABC proteins and was inhibited by orthovanadate in a profile diagnostic of ABC transporters (apparent K(I) = 12 microm). Modest stimulation (approximately 40%) was observed upon the addition of a-factor either synthetic or in native form. We also used an 8-azido-[alpha-(32)P]ATP binding and vanadate-trapping assay to examine the behavior of wild-type Ste6p and two different double mutants (G392V/G1087V and G509D/G1193D) shown previously to be mating-deficient in vivo. Both mutants displayed a diminished ability to hydrolyze ATP, with the latter uncoupled from pheromone transport. We conclude that Ste6p catalyzes ATP hydrolysis coupled to a-factor transport, which in turn promotes mating. PMID- 11389140 TI - Alteration of the specificity of malate dehydrogenase by chemical modulation of an active site arginine. AB - Malate dehydrogenase from Escherichia coli is highly specific for the oxidation of malate to oxaloacetate. The technique of site-specific modulation has been used to alter the substrate binding site of this enzyme. Introduction of a cysteine in place of the active site binding residue arginine 153 results in a mutant enzyme with diminished catalytic activity, but with K(m) values for malate and oxaloacetate that are surprisingly unaffected. Reaction of this introduced cysteine with a series of amino acid analog reagents leads to the incorporation of a range of functional groups at the active site of malate dehydrogenase. The introduction of a positively charged group such as an amine or an amidine at this position results in improved affinity for several inhibitors over that observed with the native enzyme. However, the recovery of catalytic activity is less dramatic, with less than one third of the native activity achieved with the optimal reagents. These modified enzymes do have altered substrate specificity, with alpha-ketoglutarate and hydroxypyruvate no longer functioning as alternative substrates. PMID- 11389141 TI - Structural analyses of a malate dehydrogenase with a variable active site. AB - Malate dehydrogenase specifically oxidizes malate to oxaloacetate. The specificity arises from three arginines in the active site pocket that coordinate the carboxyl groups of the substrate and stabilize the newly forming hydroxyl/keto group during catalysis. Here, the role of Arg-153 in distinguishing substrate specificity is examined by the mutant R153C. The x-ray structure of the NAD binary complex at 2.1 A reveals two sulfate ions bound in the closed form of the active site. The sulfate that occupies the substrate binding site has been translated approximately 2 A toward the opening of the active site cavity. Its new location suggests that the low catalytic turnover observed in the R153C mutant may be due to misalignment of the hydroxyl or ketone group of the substrate with the appropriate catalytic residues. In the NAD.pyruvate ternary complex, the monocarboxylic inhibitor is bound in the open conformation of the active site. The pyruvate is coordinated not by the active site arginines, but through weak hydrogen bonds to the amide backbone. Energy minimized molecular models of unnatural analogues of R153C (Wright, S. K., and Viola, R. E. (2001) J. Biol. Chem. 276, 31151-31155) reveal that the regenerated amino and amido side chains can form favorable hydrogen-bonding interactions with the substrate, although a return to native enzymatic activity is not observed. The low activity of the modified R153C enzymes suggests that precise positioning of the guanidino side chain is essential for optimal orientation of the substrate. PMID- 11389142 TI - Substitution of valine for leucine 305 in factor VIIa increases the intrinsic enzymatic activity. AB - Factor VII requires the cleavage of an internal peptide bond and the association with tissue factor (TF) to attain its fully active factor VIIa (FVIIa) conformation. The former event alone leaves FVIIa in a zymogen-like state of relatively low specific activity. We have designed a number of FVIIa mutants with the aim of mimicking the effect of TF, that is, creating molecules with increased intrinsic (TF-independent) enzymatic activity. Based on a possible structural difference between free and TF-bound FVIIa (Pike, A. C. W., Brzozowski, A. M., Roberts, S. M., Olsen, O. H., and Persson, E. (1999) Proc. Natl. Acad. Sci. U. S. A. 96, 8925--8930), we focused on the helical region encompassing residues 307 312 and residues in its spatial vicinity. For instance, FVIIa contains Phe-374 and Leu-305, whereas a Phe/Tyr residue in the position corresponding to 374 in homologous coagulation serine proteases is accompanied by Val in the position corresponding to 305. This conceivably results in a unique orientation of this helix in FVIIa. Substitution of Val for Leu-305 in FVIIa resulted in a 3--4-fold increase in the intrinsic amidolytic and proteolytic activity as compared with wild-type FVIIa, whereas the activity in complex with soluble TF remained the same. In accordance with this, L305V-FVIIa exhibited an increased rate of inhibition as compared with wild-type FVIIa, both by d-Phe-Phe-Arg-chloromethyl ketone and antithrombin III in the presence of heparin. The increased FVIIa activity upon replacement of Leu-305 by Val may be mediated by a movement of the 307--312 helix into an orientation resembling that found in factors IXa and Xa and thrombin. The corresponding shortening of the side chain of residue 374 (Phe -> Pro) had a smaller effect (about 1.5-fold increase) on the intrinsic activity of FVIIa. Attempts to increase FVIIa activity by introducing single or multiple mutations at positions 306, 309, and 312 to stabilize the 307-312 helix failed. PMID- 11389143 TI - Cloning and expression of a novel hepatitis B virus-binding protein from HepG2 cells. AB - A direct involvement of the hepatitis B virus (HBV) preS1-(21-47) sequence in virus attachment to cell membrane receptor(s) and the presence on the plasma membranes of HepG2 cells of protein(s) with receptor activity for HBV have been suggested by many previous experiments. In this study, by using a tetravalent derivative of the preS1-(21-47) sequence, we have isolated by affinity chromatography from detergent-solubilized HepG2 plasma membranes a 44-kDa protein (HBV-binding protein; HBV-BP), which was found to closely correspond to the human squamous cell carcinoma antigen 1 (SCCA1), a member of the ovalbumin family of serine protease inhibitors. Comparison of SCCA1 sequence with the sequence of the corresponding HBV-BP cDNA, cloned by polymerase chain reaction starting from RNA poly(A)(+) fractions extracted from HepG2 cells, indicated the presence of only four nucleotide substitutions in the coding region, leading to three amino acid changes. Intact recombinant HBV-BP lacked inhibitory activity for serine proteases such as alpha-chymotrypsin and trypsin but inhibited with high potency cysteine proteases such as papain and cathepsin L. Direct binding experiments confirmed the interaction of recombinant HBV-BP with the HBV preS1 domain. HepG2 cells overexpressing HBV-BP after transfection of corresponding cDNA showed a virus binding capacity increased by 2 orders of magnitude compared with untransfected cells, while Chinese hamster ovary cells, which normally do not bind to HBV, acquired susceptibility to HBV binding after transfection. Native HBV particle entry was enhanced in transfected cells. Both recombinant HBV-BP and antibodies to recombinant HBV-BP blocked virus binding and internalization in transfected cells as well as in primary human hepatocytes in a dose-dependent manner. Our findings suggest that this protein plays a major role in HBV infection. PMID- 11389144 TI - Lipid rafts exist as stable cholesterol-independent microdomains in the brush border membrane of enterocytes. AB - Glycosphingolipid/cholesterol-rich membranes ("rafts")can be isolated from many types of cells, but their existence as stable microdomains in the cell membrane has been elusive. Addressing this problem, we studied the distribution of galectin-4, a raft marker, and lactase, a protein excluded from rafts, on microvillar vesicles from the enterocyte brush border membrane. Magnetic beads coated with either anti-galectin-4 or anti-lactase antibodies were used for immunoisolation of vesicles followed by double immunogold labeling of the two proteins. A morphometric analysis revealed subpopulations of raft-rich and raft poor vesicles by the following criteria: 1) the lactase/galectin-4 labeling ratio/vesicle captured by the anti-lactase beads was significantly higher (p < or = 0.01) than that of vesicles captured by anti-galectin-4 beads, 2) subpopulations of vesicles labeled by only one of the two antibodies were preferentially captured by beads coated with the respective antibody (p < or = 0.01), 3) the average diameter of "galectin-4 positive only" vesicles was smaller than that of vesicles labeled for lactase. Surprisingly, pretreatment with methyl beta-cyclodextrin, which removed >70% of microvillar cholesterol, did not affect the microdomain localization of galectin-4. We conclude that stable, cholesterol independent raft microdomains exist in the enterocyte brush border. PMID- 11389145 TI - Secreted forms of the amyloid-beta precursor protein are ligands for the class A scavenger receptor. AB - Upon activation, platelets secrete a 120-kDa protein that competes for the binding and internalization of acetyl low density lipoproteins (AcLDL) by macrophages. From the amino-terminal amino acid sequence, amino acid composition, and immunoblot analysis, we identified the active factor in platelet secretion products as sAPP, an alpha-secretase cleavage product of the beta-amyloid precursor protein (APP), that contains a Kunitz-type protease inhibitor (KPI) domain. We showed that both sAPP751 (also called Nexin II) and sAPP695, which does not contain a KPI domain, are ligands for the class A scavenger receptor (SR A). Chinese hamster ovary cells stably transfected to express the SR-A bound and internalized 4-fold more human platelet-derived sAPP than control cells. The binding and internalization of sAPP were inhibited by the SR-A antagonist fucoidin. In addition, sAPP competed as effectively as fucoidin for SR-A-mediated cell association and degradation of (125)I-AcLDL. To determine if the KPI domain is required for the binding of sAPP to the SR-A, APP751 and APP695 were expressed in Chinese hamster ovary cells, and sAPP751 and sAPP695 purified from the medium were tested for their binding to the SR-A. sAPP751 and sAPP695 were equally effective in competing for the cell association of (125)I-AcLDL by SR-A expressing cells, demonstrating that the KPI domain is not essential for binding. We also found that sAPP751 is present in extracts of atherosclerotic lesions and that sAPP competes for the SR-A-mediated cell association of oxidized low density lipoprotein. Deletion mutagenesis indicated that a negatively charged region of APP (residues 191-264) contributes to binding to the SR-A. These results suggest that the SR-A contributes to the clearance of sAPP and that sAPP competes for the cell association of other SR-A ligands. PMID- 11389146 TI - Role of the linker region of the anion-stimulated ATPase ArsA. Effect of deletion and point mutations in the linker region. AB - The anion-stimulated ATPase ArsA in Escherichia coli consists of two homologous halves, A1 and A2, which are connected by a 40-amino acid long stretch of residues designated as the linker region. The linker region of ArsA lies in close proximity of the nucleotide-binding domain(s) of ArsA and is involved in significant conformational changes on binding of the substrates. Hence, it has been suggested earlier that the linker may play an important role in the function of ArsA. The aim of the present study was to determine the role of the linker by deletion and by site-directed mutagenesis of specific residues. Effect of deletion of the linker was determined by using the in vivo complementation approach where two halves of ArsA were co-expressed either with or without the linker region. Two co-expressed halves of ArsA conferred arsenite resistance only if the linker region was present on one of the halves. Of the six different point mutations created in the linker region, three (G284S, R290S, and D303G) were seen to drastically affect the catalytic function of ArsA. In addition, these three mutant alleles conferred arsenite sensitivity in cells carrying the wild type arsB gene. Trypsin proteolysis studies carried out with the purified proteins showed that the A1 nucleotide-binding domain in D303G protein has a conformation different from the wild type ArsA, suggesting that the linker region interacts with the nucleotide-binding domain(s) of ArsA. Based on the studies presented here, we propose that, in addition to providing flexibility, the nature of the residues themselves in the linker region is important for the conformation of the nucleotide-binding domains and for the catalytic function of ArsA. PMID- 11389147 TI - Targeted gene knockout by 2'-O-aminoethyl modified triplex forming oligonucleotides. AB - Triplex forming oligonucleotides (TFOs) are of interest because of their potential for facile gene targeting. However, the failure of TFOs to bind target sequences at physiological pH and Mg(2+) concentration has limited their biological applications. Recently, pyrimidine TFOs with 2'-O-aminoethyl (AE) substitutions were shown to have enhanced kinetics and stability of triplex formation (Cuenoud, B., Casset, F., Husken, D., Natt, F., Wolf, R. M., Altmann, K. H., Martin, P., and Moser H. E. (1998) Angew. Chem. Int. Ed. 37, 1288--1291). We have prepared psoralen-linked TFOs with varying amounts of the AE-modified residues, and have characterized them in biochemical assays in vitro, and in stability and HPRT gene knockout assays in vivo. The AE TFOs showed higher affinity for the target in vitro than a TFO with uniform 2'-OMe substitution, with relatively little loss of affinity when the assay was performed in reduced Mg(2+). Once formed they were also more stable in "physiological" buffer, with the greatest affinity and stability displayed by the TFO with all but one residue in the AE format. However, TFOs with lesser amounts of the AE modification formed the most stable triplexes in vivo, and showed the highest HPRT gene knockout activity. We conclude that the AE modification can enhance the biological activity of pyrimidine TFOs, but that extensive substitution is deleterious. PMID- 11389148 TI - Generation of a minimal alpha5beta1 integrin-Fc fragment. AB - The tertiary structure of the integrin heterodimer is currently unknown, although several predictive models have been generated. Detailed structural studies of integrins have been consistently hampered for several reasons, including the small amounts of purified protein available, the large size and conformational flexibility of integrins, and the presence of transmembrane domains and N-linked glycosylation sites in both receptor subunits. As a first step toward obtaining crystals of an integrin receptor, we have expressed a minimized dimer. By using the Fc dimerization and mammalian cell expression system designed and optimized by Stephens et al. (Stephens, P. E., Ortlepp, S., Perkins, V. C., Robinson, M. K., and Kirby, H. (2000) Cell. Adhes. Commun. 7, 377-390), a series of recombinant soluble human alpha(5)beta(1) integrin truncations have been expressed as Fc fusion proteins. These proteins were examined for their ligand binding properties and for their expression of anti-integrin antibody epitopes. The shortest functional alpha(5)-subunit truncation contained the N-terminal 613 residues, whereas the shortest beta(1)-subunit was a fragment containing residues 121-455. Each of these minimally truncated integrins displayed the antibody binding characteristics of alpha(5)beta(1) purified from human placenta and bound ligand with the same apparent affinity as the native receptor. PMID- 11389149 TI - The role of intron sequences in high level expression from CD45 cDNA constructs. AB - Consistent expression from CD45 cDNA constructs has proven difficult to achieve. Through the use of new CD45 cDNA constructs and reporter genes, the role 5', 3', and intron sequences play in CD45 expression was determined. The CD45 polyadenylation signal sequence was fully functional in a beta-galactosidase reporter construct. Furthermore, the CD45 3'-untranslated region and downstream sequences were shown to contain no negative regulatory elements. Several new CD45 cDNA constructs were designed that contain either the cytomegalovirus promoter, the leukocyte function-associated antigen (LFA-1; CD11a) promoter, or various CD45 5' regions. Neither the cytomegalovirus nor the LFA-1 promoter was capable of generating detectable levels of expression in constructs with CD45 cDNA. However, when CD45 intron sequences between exons 3 and 9 were inserted in the cDNA construct to generate a CD45 minigene, the LFA-1 promoter was able to drive reproducible, significant expression of CD45. CD45 minigenes using the CD45 5' sequences up to 19 kilobases upstream of the transcriptional start produced very little protein. The LFA-1 CD45 minigene construct produced correct cell type specific isoforms when expressed in T and B lymphocyte lines. Therefore, we conclude that the regulation of CD45 expression and cell type-specific splicing requires elements within the intron sequences. PMID- 11389150 TI - Investigation of Escherichia coli dimethyl sulfoxide reductase assembly and processing in strains defective for the sec-independent protein translocation system membrane targeting and translocation. AB - Dimethyl sulfoxide reductase is a heterotrimeric enzyme (DmsABC) localized to the cytoplasmic surface of the inner membrane. Targeting of the DmsA and DmsB catalytic subunits to the membrane requires the membrane targeting and translocation (Mtt) system. The DmsAB dimer is a member of a family of extrinsic, cytoplasmic facing membrane subunits that require Mtt in order to assemble on the membrane. We show that the MttA(2), MttB, and presumably MttA(1) but not the MttC proteins are required for targeting DmsAB to the membrane. Unlike other Mtt substrates such as trimethylamine N-oxide reductase, the soluble cytoplasmic DmsAB dimer that accumulates in the mtt deletions is very labile. Deletion of the mttA(2) or mttB genes also prevents anaerobic growth on fumarate even though fumarate reductase does not require Mtt for assembly. This was due to the lethality of membrane insertion of DmsC in the absence of the DmsAB subunits. In the absence of DmsC, DmsAB accumulates in the cytoplasm. A 45-amino acid leader on DmsA is removed during assembly. Processing does not require DmsC but does require Mtt. Translocation of DmsAB to the periplasm is not required for processing. The leader may be cleaved by a novel leader peptidase, or the long DmsA leader may traverse the membrane through the Mtt system resulting in cleavage by the periplasmic leader peptidase I followed by release of DmsA into the cytoplasm. PMID- 11389151 TI - Membrane targeting of a Rab GTPase that fails to associate with Rab escort protein (REP) or guanine nucleotide dissociation inhibitor (GDI). AB - The targeting of various Rab proteins to different subcellular compartments appears to be determined by variable amino acid sequences located upstream from geranylgeranylated cysteine residues in the C-terminal tail. All nascent Rab proteins are prenylated by geranylgeranyltransferase II, which recognizes the Rab substrate only when it is bound to Rab escort protein (REP). After prenylation, REP remains associated with the modified Rab until it is delivered to the appropriate subcellular membrane. It remains unclear whether docking of the Rab with the correct membrane is solely a function of features contained within the prenylated Rab itself (with REP serving as a "passive" carrier) or whether REP actively participates in the targeting process. To address this issue, we took advantage of a mutation in the alpha2 helix of Rab1B (i.e. Y78D) that abolishes REP and GDI interaction without disrupting nucleotide binding or hydrolysis. These studies demonstrate that replacing the C-terminal GGCC residues of Rab1B(Y78D) with a CLLL motif permits this protein to be prenylated by geranylgeranyltransferase I but not II both in cell-free enzyme assays and in transfected cells. Subcellular fractionation and immunofluorescence studies reveal that the prenylated Rab1B(Y78D)CLLL, which remains deficient in REP and GDI association is, nonetheless, delivered to the Golgi and endoplasmic reticulum (ER) membranes. When the dominant-negative S22N mutation was inserted into Rab1B CLLL, the resulting monoprenylated construct suppressed ER --> Golgi protein transport. However, when the Y78D mutation was added to the latter construct, its inhibitory effect on protein trafficking was lost despite the fact that it was localized to the ER/Golgi membrane. Therefore, protein interactions mediated by the alpha2 helical domain of Rab1B(S22N) appear to be essential for its functional interaction with components of the ER --> Golgi transport machinery. PMID- 11389152 TI - Regulation of dna replication after heat shock by replication protein a-nucleolin interactions. AB - Heat shock inhibits replicative DNA synthesis, but the underlying mechanism remains unknown. We investigated mechanistic aspects of this regulation in melanoma cells using a simian virus 40 (SV40)-based in vitro DNA replication assay. Heat shock (44 degrees C) caused a monotonic inhibition of cellular DNA replication following exposures for 5-90 min. SV40 DNA replication activity in extracts of similarly heated cells also decreased after 5-30 min of exposure, but returned to near control levels after 60-90 min of exposure. This transient inhibition of SV40 DNA replication was eliminated by recombinant replication protein A (rRPA), suggesting a regulatory process targeting this key DNA replication factor. SV40 DNA replication inhibition was associated with a transient increase in the interaction between nucleolin and RPA that peaked at 20 30 min. Because binding to nucleolin compromises the ability of RPA to support SV40 DNA replication, we suggest that the observed interaction reflects a mechanism whereby DNA replication is regulated after heat shock. The relevance of this interaction to the regulation of cellular DNA replication is indicated by the transient translocation in heated cells of nucleolin from the nucleolus into the nucleoplasm with kinetics very similar to those of SV40 DNA replication inhibition and of RPA-nucleolin interaction. Because the targeting of RPA by nucleolin in heated cells occurs in an environment that preserves the activity of several essential DNA replication factors, active processes may contribute to DNA replication inhibition to a larger degree than presently thought. RPA-nucleolin interactions may reflect an early step in the regulation of DNA replication, as nucleolin relocalized into the nucleolus 1-2 h after heat exposure but cellular DNA replication remained inhibited for up to 8 h. We propose that the nucleolus functions as a heat sensor that uses nucleolin as a signaling molecule to initiate inhibitory responses equivalent to a checkpoint. PMID- 11389157 TI - The -48 C/T polymorphism in the presenilin 1 promoter is associated with an increased risk of developing Alzheimer's disease and an increased Abeta load in brain. AB - Mutations in the presenilin 1 gene (PS1) account for the majority of early onset, familial, autosomal dominant forms of Alzheimer's disease (AD), whereas its role in other late onset forms of AD remains unclear. A -48 C/T polymorphism in the PS1 promoter has been associated with an increased genetic risk in early onset complex AD and moreover has been shown to influence the expression of the PS1 gene. This raises the possibility that previous conflicting findings from association studies with homozygosity for the PS1 intron 8 polymorphism might be the result of linkage disequilibrium with the -48 CC genotype. Here we provide further evidence of increased risk of AD associated with homozygosity for the -48 CC genotype (odds ratio=1.6). We also report a phenotypic correlation with Abeta(40), Abeta(42(43)), and total Abeta load in AD brains. The -48 CC genotype was associated with 47% greater total Abeta load (p<0.003) compared to CT + TT genotype bearers. These results suggest that the -48 C/T polymorphism in the PS1 promoter may increase the risk of AD, perhaps by altering PS1 gene expression and thereby influencing Abeta load. PMID- 11389158 TI - Peutz-Jeghers families unlinked to STK11/LKB1 gene mutations are highly predisposed to primitive biliary adenocarcinoma. AB - INTRODUCTION: Germline mutations of the STK11/LKB1 tumour suppressor gene (19p13.3) are responsible for Peutz-Jeghers syndrome (PJS), a rare genetic disorder, which is dominantly inherited. In addition to the typical hamartomatous gastrointestinal polyps and perioral pigmented lesions, PJS is also associated with the development of tumours in various sites. No specific follow up has yet been evaluated for gene carriers. Furthermore, genetic heterogeneity has been reported, which makes genetic counselling difficult. METHODS: We report here the analysis of the STK11/LKB1 locus in a series of 34 PJS families, combining the search for mutations and rearrangements in the coding sequence, allele specific expression tests, and linkage studies. RESULTS: Germline deleterious mutation of the STK11/LKB1 gene were identified in 70% of cases. The hypothesis of a second PJS locus was reinforced and PJS families could be divided into two groups on the basis of the presence or absence of an identified STK11/LKB1 alteration. Analysis of clinical data indicates that the cancer associated risk is markedly different in the two groups. PJS patients with no identified STK11/LKB1 mutation are at major risk for proximal biliary adenocarcinoma, an infrequent tumour in the general population. CONCLUSION: Up to 30% of PJS patients are caused by mutation in an unidentified gene that confers high susceptibility to cancer development. PMID- 11389159 TI - BRCA1 and BRCA2 mutation status and cancer family history of Danish women affected with multifocal or bilateral breast cancer at a young age. AB - INTRODUCTION: A small fraction of breast cancer is the result of germline mutations in the BRCA1 and BRCA2 cancer susceptibility genes. Mutation carriers frequently have a positive family history of breast and ovarian cancer, are often diagnosed at a young age, and may have a higher incidence of double or multiple primary breast tumours than breast cancer patients in general. OBJECTIVES: To estimate the prevalence and spectrum of BRCA1 and BRCA2 mutations in young Danish patients affected with bilateral or multifocal breast cancer and to determine the relationship of mutation status to family history of cancer. SUBJECTS: From the files of the Danish Breast Cancer Cooperative Group (DBCG), we selected 119 breast cancer patients diagnosed before the age of 46 years with either bilateral (n=59) or multifocal (n=61) disease. METHODS: DNA from the subjects was screened for BRCA1 and BRCA2 mutations using single strand conformation analysis (SSCA) and the protein truncation test (PTT). Observed and expected cancer incidence in first degree relatives of the patients was estimated using data from the Danish Cancer Registry. RESULTS: Twenty four mutation carriers were identified (20%), of whom 13 had a BRCA1 mutation and 11 carried a BRCA2 mutation. Two mutations in BRCA1 were found repeatedly in the material and accounted for seven of the 24 (29%) mutation carriers. The mutation frequency was about equal in patients with bilateral (22%) and multifocal breast cancer (18%). The incidence of breast and ovarian cancer was greatly increased in first degree relatives of BRCA1 and BRCA2 mutation carriers, but to a much lesser degree in relatives of non-carriers. An increased risk of cancer was also noted in brothers of non-carriers. CONCLUSIONS: A relatively broad spectrum of germline mutations was observed in BRCA1 and BRCA2 and most of the mutations are present in other populations. Our results indicate that a diagnosis of bilateral and multifocal breast cancer is predictive of BRCA1 and BRCA2 mutation status, particularly when combined with information on the patients' age at diagnosis and family history of breast/ovarian cancer. PMID- 11389160 TI - Localisation of a gene for an autosomal recessive syndrome of macrocephaly, multiple epiphyseal dysplasia, and distinctive facies to chromosome 15q26. AB - BACKGROUND: We have previously described an autosomal recessive syndrome of macrocephaly, multiple epiphyseal dysplasia (MED), and distinctive facies in a large, extended Omani family. The MED observed seems to be part of a larger malformation syndrome, since both craniofacial and central nervous system changes were present in the family. We performed a whole genome scan in this family in order to identify the gene locus for this malformation syndrome. METHODS AND RESULTS: Using homozygosity mapping, we show linkage to the telomeric region of the long arm of chromosome 15. The position of both the disease gene and the principal glycoprotein, chondroitin sulphate proteoglycan (aggrecan, AGC1) on chromosome 15q26, suggested that the aggrecan gene is a candidate for the disease in this family. However, three of the four affected children were heterozygous for a polymorphism at position 831 of the coding sequence of AGC1, providing strong evidence against its involvement. CONCLUSION: We have identified a gene locus for a recessive syndrome of macrocephaly, MED, and distinctive facies in a large Omani family. Aggrecan located on chromosome 15q26, within the critical region determined for this syndrome in this family, was excluded as a candidate gene. PMID- 11389161 TI - GATA3 abnormalities and the phenotypic spectrum of HDR syndrome. AB - We report on GATA3 analysis and the phenotypic spectrum in nine Japanese families with the HDR syndrome (hypoparathyroidism, sensorineural deafness, and renal dysplasia) (MIM 146255). Fluorescence in situ hybridisation and microsatellite analyses showed heterozygous gross deletions including GATA3 in four families. Sequence analysis showed heterozygous novel mutations in three families: a missense mutation within the first zinc finger domain at exon 4 (T823A, W275R), an unusual mutation at exon 4 (900insAA plus 901insCCT or C901AACCCT) resulting in a premature stop at codon 357 with loss of the second zinc finger domain, and a nonsense mutation at exon 6 (C1099T, R367X). No GATA3 abnormalities were identified in the remaining two families. The triad of HDR syndrome was variably manifested by patients with GATA3 abnormalities. The results suggest that HDR syndrome is primarily caused by GATA3 haploinsufficiency and is associated with a wide phenotypic spectrum. PMID- 11389162 TI - Molecular genetic heterogeneity in autosomal dominant drusen. AB - OBJECTIVE: Autosomal dominant drusen is of particular interest because of its phenotypic similarity to age related macular degeneration. Currently, mutation R345W of EFEMP1 and, in a single pedigree, linkage to chromosome 6q14 have been causally related to the disease. We proposed to investigate and quantify the roles of EFEMP1 and the 6q14 locus in dominant drusen patients from the UK and USA. DESIGN: Molecular genetic analysis. PARTICIPANTS: Ten unrelated families and 17 young drusen patients. MAIN OUTCOME MEASURES: Exons 1 and 2 of EFEMP1 were characterised by 5' rapid amplification of cDNA ends and direct sequencing. Exons 1-12 of EFEMP1 were then investigated for mutation by direct sequencing. A HpaII restriction digest test was constructed to detect the EFEMP1 R345W mutation. Marker loci spanning the two dominant drusen linked loci were used to generate haplotype data. RESULTS: Only seven of the 10 families (70%) and one of the 17 sporadic patients (6%) had the R345W mutation. The HpaII restriction digest test was found to be a reliable and quick method for detecting this. No other exonic or splice site mutation was identified. Of the three families without EFEMP1 mutation, two were linked to the 2p16 region. CONCLUSIONS: EFEMP1 R345W accounts for only a proportion of the dominant drusen phenotype. Importantly, other families linked to chromosome 2p16 raise the possibility of EFEMP1 promoter sequence mutation or a second dominant drusen gene at this locus. Preliminary haplotype data suggest that the disease gene at the 6q14 locus is responsible for only a minority of dominant drusen cases. PMID- 11389164 TI - Mutations in the glycine receptor alpha1 subunit (GLRA1) gene in hereditary hyperekplexia pedigrees: evidence for non-penetrance of mutation Y279C. PMID- 11389165 TI - Ellis-van Creveld syndrome resulting from segmental uniparental disomy of chromosome 4. PMID- 11389166 TI - Nijmegen breakage syndrome in a Dutch patient not resulting from a defect in NBS1. PMID- 11389167 TI - Hair roots as the ideal source of mRNA for genetic testing. PMID- 11389168 TI - Two further cases of WHS with unbalanced de novo translocation t(4;8) characterised by CGH and FISH. PMID- 11389169 TI - Differential expression of mRNAs for sialyltransferase isoenzymes induced in the hippocampus of mouse following kindled seizures. AB - Sialic acids play important roles in various biological functions. In the brain, evidence suggests that sialylation of glycoproteins and glycolipids affects neural plasticity. While the 18 sialyltransferase isoenzymes (STs) identified to date synthesize individual sialyl-oligosaccharide structures, they each exhibit activity toward more than one substrate and can overlap in their specificity. Therefore, the distribution of STs is a secondary factor in the study of specific sialylation. Here, seven STs; ST3Gal I-IV, ST8Sia IV, ST6Gal I and ST6GalNAc II, the expressions of which were identified in the adult hippocampus by RT-PCR, showed diverse localization patterns in the hippocampus on in situ hybridization, suggesting that the individual cells expressed relevant STS: Furthermore, to assay activity-related changes in ST expression, we used amygdaloid-kindling among models of neural plasticity. Differential expression of the STs participating in the kindling, notably, up-regulation of ST3Gal IV and ST6GalNAc II mRNAs, and down-regulation of ST3Gal I and ST8Sia IV mRNAs, were observed in the hippocampus following kindled seizures. These results indicate that ST expressions are regulated by physiological activity and may play a role in neural plasticity. PMID- 11389170 TI - Long-term potentiation in the dentate gyrus of the rat hippocampus is accompanied by brain-derived neurotrophic factor-induced activation of TrkB. AB - A role for neurotrophic factors, in particular brain-derived neurotrophic factor (BDNF), in modulating synaptic plasticity in the adult brain has been described in recent years by several laboratories. A great deal of emphasis has been placed on establishing its precise role in the expression of long-term potentiation (LTP) in the hippocampus. Here we attempt to address this question by investigating, first, its release following induction of LTP in perforant path granule cell synapses and, second, the signalling events which follow activation of the BDNF receptor, TrkB, in the presynaptic terminal. We report that BDNF release is increased from slices of dentate gyrus following tetanic stimulation of the perforant path and that TrkB activation is increased in synaptosomes prepared from tetanized dentate gyrus. These changes are accompanied by increased activation of one member of the family of mitogen-activated protein kinases, extracellular signal-regulated kinase (ERK) and the data indicate that these events play a role in modulating release of glutamate from perforant path-granule cell synapses, because the Trk inhibitor K252a and the ERK inhibitor, UO126, both inhibited the BDNF-induced enhancement of release. We propose that the increase in phosphorylation of the transcription factor cAMP response element binding protein and in protein synthesis might underlie the more persistent components of LTP in dentate gyrus. PMID- 11389171 TI - Glatiramer acetate inhibition of tumor necrosis factor-alpha-induced RANTES expression and release from U-251 MG human astrocytic cells. AB - Glatiramer acetate is an approved drug for the treatment of multiple sclerosis (MS). RANTES is a beta-family chemokine that manifests chemoattractant activity for T lymphocytes and monocytes/macrophages implicated in the pathogenesis of MS lesions. However, the effect of glatiramer acetate on the regulation of RANTES secretion in glial cells is unknown. In the present study, we demonstrate for the first time that treatment of human U-251 MG astrocytic cells with glatiramer acetate blocks tumor necrosis factor-alpha (TNF-alpha)-induced RANTES mRNA and protein in a dose- and time-dependent manner. This effect is attributed to inhibition of transcription and a 40% decrease in transcript stability. Furthermore, our electrophoretic mobility shift assays of nuclear extracts from TNF-alpha-treated cells reveal an increase in DNA-binding activity specific for the nuclear factor-kappa B (NF-kappaB) binding site, in the 5'-flanking promoter region of the human RANTES gene, and that this increase in NF-kappaB binding activity is prevented by pretreatment with glatiramer acetate or the NF-kappaB inhibitors. These findings suggest that glatiramer acetate may exert its therapeutic effect in MS partially through inhibiting NF-kappaB activation and chemokine production. PMID- 11389172 TI - [(3)H](2S,4R)-4-Methylglutamate: a novel ligand for the characterization of glutamate transporters. AB - [(3)H](2S,4R)-4-Methylglutamate ([(3)H]4MG), used previously as a ligand for low affinity kainate receptors, was employed to establish a binding assay for glutamate transporters (GluTs), as 4MG has also been shown to have affinity for the glial GluTs, GLT1 and GLAST. In rat brain membrane homogenates in the presence of Na(+) ions at 4 degrees C, specific binding of [(3)H]4MG was rapid and saturable (t(1/2) approximately 15 min), representing > 90% of total binding. Dissociation of [(3)H]4MG occurred in a biphasic manner, however, saturation studies and Scatchard analysis indicated a single site of binding (n(H) = 0.85) and a K(d) of 6.2 +/- 0.8 microM with a B(max) of 111.8 +/- 23.8 pmol/mg protein. Specific binding of [(3)H]4MG was Na(+)-dependent and inhibited by K(+) and HCO(3 ). Pharmacological inhibition with compounds acting at GluTs revealed that Glu, D and L-aspartate, L-serine-O-sulfate and Ltrans-pyrrolidine-2,4-dicarboxylate fully displaced specific binding. Drugs having preferential affinity for GLT1, kainate, dihydrokainate and Lthreo-3-methylglutamate, all inhibited approximately 40% of specific binding. The inhibition pattern of L-serine-O-sulfate in the presence of a saturating concentration of dihydrokainate was suggestive of [(3)H]4MG also labelling GLAST. 6-Cyano-7-nitroquinoxaline, a kainate receptor antagonist, and a range of Glu receptor agonists and antagonists failed to significantly inhibit [(3)H]4MG binding. The pharmacological profile of binding of [(3)H]4MG resembled that found for [(3)H]D-aspartate, a ligand specific for GluTs, reinforcing the hypothesis that [(3)H]4MG was labelling GluTs in this assay. Together, these data illustrate the development of an efficient, economic binding assay that is suitable for the characterization of different subtypes of GLuTs. PMID- 11389173 TI - Stromal cell-derived factor-1alpha induces astrocyte proliferation through the activation of extracellular signal-regulated kinases 1/2 pathway. AB - Stromal cell-derived factor-1 (SDF-1), the ligand of the CXCR4 receptor, is a chemokine involved in chemotaxis and brain development that also acts as co receptor for HIV-1 infection. We previously demonstrated that CXCR4 and SDF 1alpha are expressed in cultured type-I cortical rat astrocytes, cortical neurones and cerebellar granule cells. Here, we investigated the possible functions of CXCR4 expressed in rat type-I cortical astrocytes and demonstrated that SDF-1alpha stimulated the proliferation of these cells in vitro. The proliferative activity induced by SDF-1alpha in astrocytes was reduced by PD98059, indicating the involvement of extracellular signal-regulated kinases (ERK1/2) in the astrocyte proliferation induced by CXCR4 stimulation. This observation was further confirmed showing that SDF-1alpha treatment selectively activated ERK1/2, but not p38 or stress-activated protein kinase/c-Jun N-terminal kinase (SAPK/JNK). Moreover, both astrocyte proliferation and ERK1/2 phosphorylation, induced by SDF-1alpha, were inhibited by pertussis toxin (PTX) and wortmannin treatment indicating the involvement of a PTX sensitive G-protein and of phosphatidyl inositol-3 kinase in the signalling of SDF-1alpha. In addition, Pyk2 activation represent an upstream components for the CXCR4 signalling to ERK1/2 in astrocytes. To our knowledge, this is the first report demonstrating a proliferative effect for SDF-1alpha in primary cultures of rat type-I astrocytes, and showing that the activation of ERK1/2 is responsible for this effect. These data suggest that CXCR4/SDF-1 should play an important role in physiological and pathological glial proliferation, such as brain development, reactive gliosis and brain tumour formation. PMID- 11389175 TI - D1-D2 dopamine receptor interaction within the nucleus accumbens mediates long loop negative feedback to the ventral tegmental area (VTA). AB - The objective of the present study was to examine the effects of perfusion of dopamine (DA) D1- and D2-like receptor agonists in the nucleus accumbens (ACB) on the long-loop negative feedback regulation of mesolimbic somatodendritic DA release in the ventral tegmental area (VTA) of Wistar rats employing ipsilateral dual probe in vivo microdialysis. Perfusion of the ACB for 60 min with the D1 like receptor agonist SKF 38393 (SKF, 1-100 microM) dose-dependently reduced the extracellular levels of DA in the ACB, whereas the extracellular levels of DA in the VTA were not changed. Similarly, application of the D2-like receptor agonist quinpirole (Quin, 1-100 microM) through the microdialysis probe in the ACB reduced the extracellular levels of DA in the ACB in a concentration-dependent manner, whereas extracellular levels of DA in the VTA were not altered. Co application of SKF (100 microM) and Quin (100 microM) produced concomitant reductions in the extracellular levels of DA in the ACB and VTA. The reduction in extracellular levels of DA in the ACB and VTA produced by co-infusion of SKF and Quin was reversed in the presence of either 100 microM SCH 23390 (D1-like antagonist) or 100 microM sulpiride (D2-like antagonist). Overall, the results suggest that (a) activation of dopamine D1- or D2-like receptors can independently regulate local terminal DA release in the ACB, whereas stimulation of both subtypes is required for activation of the negative feedback pathway to the VTA. PMID- 11389174 TI - Constitutive expression of functional GABA(B) receptors in mIL-tsA58 cells requires both GABA(B(1)) and GABA(B(2)) genes. AB - Studies of gamma-aminobutyric acid (GABA)(B) receptor function in heterologous cell systems have suggested that expression of two distinct seven transmembrane G protein coupled receptor subunits is necessary for receptor activation and signal transduction. Some results suggest that both receptor proteins must be inserted into the plasma membrane to create heterodimers; however, it is possible that subunit monomers or homodimers are functional in cells which constitutively express GABA(B) receptors. A new pituitary intermediate lobe melanotrope cell clone (mIL tsA58) has been isolated which constitutively expresses GABA(B), D(2) and corticotrophin releasing factor receptors. Here, we report on characterization of the GABA(B) receptors. Solution hybridization-nuclease protection assays reveal the presence of GABA(B(1)) and GABA(B(2)) transcripts. Western blots show GABA(B(1a)) and one of two GABA(B(2)) proteins. Addition of the GABA(B) agonist baclofen to cultured mIL-tsA58 (mIL) cells inhibits high voltage activated Ca(2+) channels, as measured by agonist-induced inhibition of the K(+)-depolarization-stimulated increase in Ca(2+) influx. CGP55845, a GABA(B) antagonist, blocks the response to baclofen. Knockdown of either GABA(B(1)) or GABA(B(2)) subunits with selective antisense oligodeoxynucleotides reduced GABA(B) protein levels and completely abolished the GABA(B) receptor response in the mIL cells. Taken together, these results indicate that functionally active GABA(B) receptors in mIL cells require the constitutive expression of both GABA(B) genes. This is a physiologic validation of results from recombinant overexpression in naive cells and shows that the mIL cell line is a useful model for studying GABA(B) receptor expression, regulation and function. PMID- 11389176 TI - Genomic structure, developmental distribution and functional properties of the chicken P2X(5) receptor. AB - We report here the cloning of a chicken cDNA (402 aa) showing high sequence similarity to the previously cloned rat and human P2X(5) receptors (67 and 69%, respectively). The chicken P2X(5) subunit is encoded by a gene composed of 12 translated exons, which shows conserved genomic structure with mammalian P2X genes. In HEK-293 cells heterologously expressing chicken P2X(5) receptors, ATP activates a current that desensitizes in a way that is dependent on the presence of extracellular divalent cations. ATP and 2-methylthio ATP are equipotent agonists (EC(50) approximately 2 microM) and suramin and pyridoxal 5-phosphate-6 azophenyl-2',4'-disulfonic acid are potent antagonists. Additionally, reversal potential measurements indicate that chicken P2X(5) is permeable not only to cations but also to chloride (P(Cs+)/P(Cl-) approximately 1.9), as has been described for native P2X receptor mediated responses in embryonic chicken skeletal muscle. mRNA distribution of chicken P2X(5) was determined by in situ hybridization analysis in both whole embryos and on tissue slices of heart and skeletal muscle. Our results suggest that chicken P2X(5) receptors are expressed in developing muscle and might play a role in early muscle differentiation. PMID- 11389177 TI - gamma-Aminobutyric acid(A) receptor subunit expression predicts functional changes in hippocampal dentate granule cells during postnatal development. AB - Profound alterations in the function of GABA occur over the course of postnatal development. Changes in GABA(A) receptor expression are thought to contribute to these differences in GABAergic function, but how subunit changes correlate with receptor function in individual developing neurons has not been defined precisely. In the current study, we correlate expression of 14 different GABA(A) receptor subunit mRNAs with changes in the pharmacological properties of the receptor in individual hippocampal dentate granule cells over the course of postnatal development in rat. We demonstrate significant developmental differences in GABA(A) receptor subunit mRNA expression, including greater than two-fold lower expression of alpha1-, alpha4- and gamma2-subunit mRNAs and 10 fold higher expression of alpha5-mRNA in immature compared with adult neurons. These differences correlate both with regional changes in subunit protein level and with alterations in GABA(A) receptor function in immature dentate granule cells, including two-fold higher blockade by zinc and three-fold lower augmentation by type-I benzodiazepine site modulators. Further, we find an inverse correlation between changes in GABA(A) receptor zinc sensitivity and abundance of vesicular zinc in dentate gyrus during postnatal development. These findings suggest that developmental differences in subunit expression contribute to alterations in GABA(A) receptor function during postnatal development. PMID- 11389178 TI - Nitric oxide regulates adenylyl cyclase activity in rat striatal membranes. AB - The regulation of adenylyl cyclase activity by nitric oxide (NO) was studied in rat (Sprague-Dawley) striatal membranes. Three chemically distinct NO donors attenuated forskolin-stimulated activity but did not alter basal activity. Maximum inhibition resulted in a 50% decrease in forskolin-stimulated activity, consistent with the presence of multiple isoforms of adenylyl cyclase and our previous findings that only the forskolin-stimulated activity of the type-5 and 6 isoform family of enzymes is inhibited by NO. To monitor primarily the type-5 isoform, we examined the ability of NO donors to attenuate D(1)-agonist stimulated adenylyl cyclase activity. Under those conditions, complete inhibition was observed. The data indicate that NO attenuates neuromodulator-stimulated cAMP signaling in the striatum. PMID- 11389179 TI - Up-regulation of hippocampal glutamate transport during chronic treatment with sodium valproate. AB - Excessive glutamatergic neurotransmission has been implicated in some neurodegenerative disorders. It would be of value to know whether glutamate transport, which terminates the glutamate signal, can be up-regulated pharmacologically. Here we show that chronic treatment of rats with the anti epileptic drug sodium valproate (200 mg or 400 mg/kg bodyweight, twice per day for 90 days) leads to a dose-dependent increase in hippocampal glutamate uptake capacity as measured by uptake of [(3)H]glutamate into proteoliposomes. The level of glutamate transporters EAAT1 and EAAT2 in hippocampus also increased dose dependently. No effect of sodium valproate on glutamate transport was seen in frontal or parietal cortices or in cerebellum. The hippocampal levels of glial fibrillary acidic protein and glutamine synthetase were unaffected by valproate treatment, whereas the levels of synapsin I and phosphate-activated glutaminase were reduced by valproate treatment, suggesting that the increase in glutamate transporters was not caused by astrocytosis or increased synaptogenesis. A direct effect of sodium valproate on the glutamate transporters could be excluded. The results show that hippocampal glutamate transport is an accessible target for pharmacological intervention and that sodium valproate may have a role in the treatment of excitotoxic states in the hippocampus. PMID- 11389180 TI - The role of putative intragenic control elements in c-fos regulation by calcium and growth factor signalling pathways. AB - Sequences in the transcribed region of the c-fos gene have been suggested to control c-fos induction following exposure of cells to mitogens or stimuli that increase intracellular calcium concentrations. Using a mutational analysis we show that putative regulatory elements present in the first intron of the human c fos gene and the fos-intragenic-regulatory-element (FIRE) are not required for c fos regulation by growth factor and calcium signalling pathways in AtT20 and PC12 cells. Removal of the c-fos first intron and the FIRE did not increase the basal level of c-fos mRNA and only moderately reduced the magnitude of calcium-induced transcription mediated by either the entire c-fos promoter or the cAMP response element (CRE). Intragenic mutations did not affect serum response element (SRE) dependent gene expression induced by calcium signals but caused a superinduction of c-fos expression in nerve growth factor-stimulated PC12 cells. These results indicate that c-fos promoter elements, rather than intragenic sequences, are the principal targets of transcription-regulating signalling pathways. This suggests that CRE- and SRE-bound activators of transcription initiation may also enhance, in a signal-dependent manner, c-fos transcript elongation beyond promoter proximal pause sites. PMID- 11389181 TI - Identification of a basolateral membrane targeting signal within the cytoplasmic domain of myelin/oligodendrocyte glycoprotein. AB - Oligodendrocytes possess two distinct membrane compartments--uncompacted plasma membrane (cell body, processes) and compact myelin. Specific targeting mechanisms must exist to establish and maintain these membrane domains. Polarized epithelial cells have the best characterized system for targeting components to apical and basolateral compartments. Since oligodendrocytes arise from neuroepithelial cells, we investigated whether they might utilize targeting paradigms similar to polarized epithelial cells. Myelin/oligodendrocyte glycoprotein (MOG) is a transmembrane Ig-like molecule restricted to uncompacted oligodendroglial plasma membrane. We stably expressed MOG in Madin-Darby canine kidney (MDCK) Type II epithelial cells, which have been extensively used in protein-targeting studies. Data from surface biotinylation assays and confocal microscopy revealed that MOG sorts exclusively to the basolateral membrane of MDCK cells. Expression vectors containing progressive truncations of MOG from the cytoplasmic C-terminus were expressed in MDCK cells to localize basolateral sorting signals. A loss of only four C-terminal residues results in some MOG expression at the apical surface. More strikingly, removal of the C-terminal membrane associated hydrophobic domain from MOG results in complete loss of basolateral sorting and specific targeting to the apical membrane. These data suggest that myelinating oligodendrocytes may utilize a sorting mechanism similar to that of polarized epithelia. PMID- 11389183 TI - Testosterone-mediated neuroprotection through the androgen receptor in human primary neurons. AB - Estrogen is an active neuroprotectant and is presently investigated as a potential therapy against Alzheimer's disease for women. To determine if male hormones could also be neuroprotective, we investigated the effect of testosterone, methyltestosterone, and epitestosterone at physiological concentrations on primary cultures of human neurons induced to undergo apoptosis by serum deprivation. Serum deprivation significantly induces neuronal apoptosis in a protracted fashion. As expected, physiological concentrations of 17-beta estradiol and transcriptionally inactive 17-alpha-estradiol protect neurons against apoptosis. Similar to 17-beta-estradiol, physiological concentrations of testosterone are also neuroprotective. Androgen receptors are present at 8 +/- 2 fmol/mg protein in the neuron cultures. The non-aromatizable androgen, mibolerone, is also neuroprotective and aromatase inhibitor, 4-androsten-4-OL 3,17-dione, does not prevent testosterone-mediated neuroprotection. In contrast, anti-androgen, flutamide, eliminates testosterone-mediated neuroprotection. Testosterone analog, methyltestosterone, showed androgen receptor-dependent neuroprotection that was delayed in time indicating that a metabolite may be the active agent. The endogenous anti-androgen, epitestosterone, also showed a slight neuroprotective effect but not through the androgen receptor. These results indicate that androgens induce neuroprotection directly through the androgen receptor. These data suggest that androgens may also be of therapeutic value against Alzheimer's disease in aging males. PMID- 11389182 TI - Similarities and differences in the neuronal death processes activated by 3OH kynurenine and quinolinic acid. AB - 3OH-Kynurenine and quinolinic acid are tryptophan metabolites able to cause, at relatively elevated concentrations, neuronal death in vitro and in vivo. In primary cultures of mixed cortical cells, the minimal concentration of these compounds leading to a significant degree of neurotoxicity decreased from 100 to 1 microM, when the exposure time was prolonged from 24 to 72 h. NMDA receptor antagonists and inhibitors of nitric oxide synthase or poly(ADP-ribose) polymerase reduced quinolinic acid, but not 3OH-kynurenine toxicity. In contrast, scavengers of free radicals, caspase inhibitors and cyclosporin preferentially reduced 3OH-kynurenine neurotoxicity. These observations suggest that quinolinic acid causes necrosis, whereas 3OH-kynurenine-exposed neurons primarily die in apoptosis. In line with this possibility, we found that ATP levels decreased more rapidly in quinolinate- than in 3OH-kynurenine-exposed cultures and that poly(ADP ribose) polymer, the product of poly(ADP-ribose) polymerase activity, was more abundant in the nuclei of quinolinic acid than in those of 3OH-kynurenine-exposed neurons. Because minor changes in the physiological concentrations of 3OH kynurenine and quinolinic acid may cause neuronal death, our data suggest that these metabolites play a key role in the pathogenesis of several neurological disorders. PMID- 11389184 TI - Functional expression of M(1), M(3) and M(5) muscarinic acetylcholine receptors in yeast. AB - The goal of this study was to functionally express the three G(q)-coupled muscarinic receptor subtypes, M(1), M(3) and M(5), in yeast (Saccharomyces cerevisiae). Transformation of yeast with expression constructs coding for the full-length receptors resulted in very low numbers of detectable muscarinic binding sites (B(max) < 5 fmol/mg). Strikingly, deletion of the central portion of the third intracellular loops of the M(1), M(3) and M(5) muscarinic receptors resulted in dramatic increases in B(max) values (53-214 fmol/mg). To monitor productive receptor/G-protein coupling, we used specifically engineered yeast strains that required agonist-stimulated receptor/G-protein coupling for cell growth. These studies showed that the shortened versions of the M(1), M(3) and M(5) receptors were unable to productively interact with the endogenous yeast G protein alpha-subunit, Gpa1p, or a Gpa1 mutant subunit that contained C-terminal mammalian Galpha(s) sequence. In contrast, all three receptors gained the ability to efficiently couple to a Gpa1/Galpha(q) hybrid subunit containing C-terminal mammalian Galpha(q) sequence, indicating that the M(1), M(3) and M(5) muscarinic receptors retained proper G-protein coupling selectivity in yeast. This is the first study to report the expression of muscarinic receptors in a coupling competent form in yeast. The strategy described here, which involves structural modification of both receptors and co-expressed G proteins, should facilitate the functional expression of other classes of G protein-coupled receptors in yeast. PMID- 11389185 TI - Evidence against an essential role of endogenous brain dopamine in methamphetamine-induced dopaminergic neurotoxicity. AB - The present studies examined the role of endogenous dopamine (DA) in methamphetamine (METH)-induced dopaminergic neurotoxicity while controlling for temperature-related neuroprotective effects of the test compounds, reserpine and alpha-methyl-p-tyrosine (AMPT). To determine if the vesicular pool of DA was essential for the expression of METH-induced DA neurotoxicity, reserpine (3 mg/kg, given iintraperitoneally 24-26 h prior to METH) was given prior to a toxic dose regimen of METH. Despite severe striatal DA deficits during the period of METH exposure, mice treated with reserpine prior to METH developed long-term reductions in striatal DA axonal markers, suggesting that vesicular DA stores were not crucial for the development of METH neurotoxicity, but leaving open the possibility that cytoplasmic DA might be involved. To evaluate this possibility, cytoplasmic DA stores were depleted with AMPT prior to METH administration. When this study was carried out at 28 degrees C, complete neuroprotection was observed, likely due to lingering effects on core temperature because when the same study was repeated at 33 degrees C (to eliminate AMPT's hypothermic effect in METH-treated animals), the previously observed neuroprotection was no longer evident. In the third and final set of experiments, mice were pretreated with a combination of reserpine and AMPT, to deplete both vesicular and cytoplasmic DA pools, and to reduce striatal DA levels to negligible values during the period of METH administration (< 0.05%). When core temperature differences were eliminated by raising ambient temperature, METH-induced DA neurotoxic changes were evident in mice pretreated with reserpine and AMPT. Collectively, these findings bring into question the view that endogenous DA plays an essential role in METH-induced DA neurotoxicity. PMID- 11389186 TI - Inhibitors of Na(+)/H(+) and Na(+)/Ca(2+) exchange potentiate methamphetamine induced dopamine neurotoxicity: possible role of ionic dysregulation in methamphetamine neurotoxicity. AB - Although the neurotoxic potential of methamphetamine (METH) is well established, underlying mechanisms have yet to be identified. In the present study, we sought to determine whether ionic dysregulation was a feature of METH neurotoxicity. In particular, we reasoned that if METH impairs the function of Na(+)/H(+) and/or Na(+)/Ca(2+) antiporters by compromising the inward Na(+) gradient [via prolonged DA transporter (DAT) activation and Na(+)/K(+) ATPase inhibition], then amiloride (AMIL) and other inhibitors of Na(+)/H(+) and/or Na(+)/Ca(2+) exchange would potentiate METH neurotoxicity. To test this hypothesis, mice were treated with METH alone or in combination with AMIL or one of its analogs; 1 week later, the animals were killed for studies of dopamine (DA) neuronal integrity. AMIL markedly potentiated the toxic effect of METH on DA neurons. Potentiation was not caused by increased core temperature, enhanced DAT activity or higher METH brain levels. The DAT inhibitor, WIN-35,428, protected completely against METH-induced DA neurotoxicity in AMIL pretreated animals, suggesting that the potentiating effects of AMIL require a METH/DAT interaction. Findings with METH and AMIL were extended to six other AMIL analogs (MIA, EIPA, DIMA, BENZ, BEP, DiCBNZ), another species (rats), and neuronal type (5-HT neurons). These results support the notion that ionic dysregulation may play a role in METH neurotoxicity. PMID- 11389187 TI - Glutathione S-transferase Pi is a dopamine-inducible suppressor of dopamine induced apoptosis in PC12 cells. AB - The finding that the neurotransmitter dopamine induces apoptosis in neurons implies the existence of a cellular mechanism by which dopaminergic neurons protect themselves from dopamine-induced apoptosis. By profiling the expression of a number of genes in differentiating PC12 cells which exhibit elevated levels of dopamine biosynthesis, we found that expression of glutathione S-transferase class Pi (GSTp) mRNA was selectively up-regulated. Interestingly, dopamine added to the culture medium of PC12 cells also augmented their expression of GSTp mRNA. Suppression of GSTp expression by transfection of its antisense expression vector augmented dopamine-induced apoptosis of PC12 cells. Conversely, overexpression of GSTp made the resultant PC12 transfectants highly resistant to dopamine-induced apoptosis. Transfection of the antisense or sense GSTp expression vectors also resulted in corresponding augmentation or suppression of dopamine-induced activation of cell-associated Jun-N-terminal kinase (JNK), which has been suggested to mediate dopamine-induced apoptosis in neuronal cells. These results indicate that GSTp is a dopamine-inducible suppressor of dopamine-induced apoptosis in PC12 cells, and suggest that this activity is exerted through inhibition of JNK activity. PMID- 11389188 TI - Tau-66: evidence for a novel tau conformation in Alzheimer's disease. AB - We have characterized a novel monoclonal antibody, Tau-66, raised against recombinant human tau. Immunohistochemistry using Tau-66 reveals a somatic neuronal stain in the superior temporal gyrus (STG) that is more intense in Alzheimer's disease (AD) brain than in normal brain. In hippocampus, Tau-66 yields a pattern similar to STG, except that neurofibrillary lesions are preferentially stained if present. In mild AD cases, Tau-66 stains plaques lacking obvious dystrophic neurites (termed herein 'diffuse reticulated plaques') in STG and the hippocampus. Enzyme-linked immunosorbent assay (ELISA) analysis reveals that Tau-66 is specific for tau, as there is no cross-reactivity with MAP2, tubulin, Abeta(1-40), or Abeta(1-42), although Tau-66 fails to react with tau or any other polypeptide on western blots. The epitope of Tau-66, as assessed by ELISA testing of tau deletion mutants, appears discontinuous, requiring residues 155-244 and 305-314. Tau-66 reactivity exhibits buffer and temperature sensitivity in an ELISA format and is readily abolished by SDS treatment. Taken together these lines of evidence indicate that the Tau-66 epitope is conformation dependent, perhaps involving a close interaction of the proline-rich and the third microtubule-binding regions. This is the first indication that tau can undergo this novel folding event and that this conformation of tau is involved in AD pathology. PMID- 11389189 TI - Proinflammatory cytokines promote glial heme oxygenase-1 expression and mitochondrial iron deposition: implications for multiple sclerosis. AB - Proinflammatory cytokines, pathological iron deposition, and oxidative stress have been implicated in the pathogenesis of multiple sclerosis (MS) and experimental autoimmune encephalomyelitis (EAE). HO-1 mRNA levels and mitochondrial uptake of [(55)Fe]Cl(3)-derived iron were measured in rat astroglial cultures exposed to interleukin-1beta (IL-1beta) or tumor necrosis factor-alpha (TNF-alpha) alone or in combination with the heme oxygenase-1 (HO-1) inhibitors, tin mesoporphyrin (SnMP) or dexamthasone (DEX), or interferon beta1b (INF-beta). HO-1 expression in astrocytes was evaluated by immunohistochemical staining of spinal cord tissue derived from MS and control subjects. IL-1beta or TNF-alpha promoted sequestration of non-transferrin-derived (55)Fe by astroglial mitochondria. HO-1 inhibitors, mitochondrial permeability transition pore (MTP) blockers and antioxidants significantly attenuated cytokine-related mitochondrial iron sequestration in these cells. IFN-beta decreased HO-1 expression and mitochondrial iron sequestration in IL-1beta- and TNF-alpha-challenged astroglia. The percentage of astrocytes coexpressing HO-1 in affected spinal cord from MS patients (57.3% +/- 12.8%) was significantly greater (p < 0.05) than in normal spinal cord derived from controls subjects (15.4% +/- 8.4%). HO-1 is over expressed in MS spinal cord astroglia and may promote mitochondrial iron deposition in MS plaques. In MS, IFN-beta may attenuate glial HO-1 gene induction and aberrant mitochondrial iron deposition accruing from exposure to proinflammatory cytokines. PMID- 11389190 TI - Pharmacological and functional characterization of muscarinic receptor subtypes in developing oligodendrocytes. AB - This study focused on the molecular and pharmacological characterization of muscarinic acetylcholine receptors expressed by progenitors and differentiated oligodendrocytes. We also analyzed the role of muscarinic receptors in regulating downstream signal transduction pathways and the functional significance of receptor expression in oligodendrocytes. RT-PCR analysis revealed the expression of transcripts for M3, and to a lesser extent M4, followed by M1, M2 and M5 receptor subtypes in both progenitors and differentiated oligodendrocytes. Competition binding experiments using [(3)H]N-methylscopolamine and several antagonists, as well as inhibition of carbachol-mediated phosphoinositide hydrolysis, showed that M3 is the main subtype expressed in these cells. In progenitors the activation of p42/44-mitogen-activated protein kinase (MAPK) and cAMP-response element binding protein (CREB) as well as c-fos mRNA expression were blocked by the M3 relatively selective antagonist, 4-DAMP, and its irreversible analogue, 4-DAMP-mustard. Carbachol increased proliferation of progenitors, an effect prevented by atropine and 4-DAMP, as well as by the MAPK kinase inhibitor PD98059. These results indicate that carbachol modulates oligodendrocyte progenitor proliferation through M3 receptors, involving activation of a MAPK signaling pathway. Receptor density and phosphoinositide hydrolysis are down-regulated during oligodendrocyte differentiation. Functional consequences of these events are a reduction in carbachol-stimulated p42/44(MAPK) and CREB phosphorylation, as well as induction of c-fos. PMID- 11389191 TI - Analysis of the mutant Drosophila N-ethylmaleimide sensitive fusion-1 protein in comatose reveals molecular correlates of the behavioural paralysis. AB - NEM-sensitive fusion protein (NSF) is an ATPase required for many intracellular membrane trafficking steps. Recent studies have suggested that NSF alters the conformation of the SNAP receptors (SNAREs) to permit their interaction, or to uncouple them after they interact. Most organisms have a single NSF gene product but Drosophila express two highly related isoforms, dNSF-1 and dNSF-2. dNSF-1 is encoded by the gene comatose (comt), first identified as the locus of a temperature-sensitive paralytic mutation. Here we show that dNSF-1 is most abundant in the nervous system and can be detected in larval and adult CNS. Subcellular fractionation revealed that dNSF-1 was enriched in a vesicle fraction along with the synaptic vesicle protein synaptotagmin. comt flies maintained at the non-permissive temperature rapidly accumulate sodium dodecyl sulfate (SDS) resistant SNARE complexes at the restrictive temperature, with concomitant translocation of dNSF-1 from cytosol and membrane fractions into a Triton X-100 insoluble fraction. The long recovery of comt flies after heat shock induced paralysis correlated with the irreversibility of this translocation. Interestingly, while dNSF-1 also translocates in comt(TP7) larvae, there is no associated neurophysiological phenotype at the neuromuscular junction (nmj) or accumulation of SDS-resistant complexes in the CNS. Together, these results suggest that dNSF-1 is required for adult neuronal function, but that in the larval nmj function may be maintained by other isoforms. PMID- 11389192 TI - Brain ischemia and reperfusion activates the eukaryotic initiation factor 2alpha kinase, PERK. AB - Reperfusion after global brain ischemia results initially in a widespread suppression of protein synthesis in neurons, which persists in vulnerable neurons, that is caused by the inhibition of translation initiation as a result of the phosphorylation of the alpha-subunit of eukaryotic initiation factor 2 (eIF2alpha). To identify kinases responsible for eIF2alpha phosphorylation [eIF2alpha(P)] during brain reperfusion, we induced ischemia by bilateral carotid artery occlusion followed by post-ischemic assessment of brain eIF2alpha(P) in mice with homozygous functional knockouts in the genes encoding the heme regulated eIF2alpha kinase (HRI), or the amino acid-regulated eIF2alpha kinase (GCN2). A 10-fold increase in eIF2alpha(P) was observed in reperfused wild-type mice and in the HRI-/- or GCN2-/- mice. However, in all reperfused groups, the RNA-dependent protein kinase (PKR)-like endoplasmic reticulum eIF2alpha kinase (PERK) exhibited an isoform mobility shift on SDS-PAGE, consistent with the activation of the kinase. These data indicate that neither HRI nor GCN2 are required for the large increase in post-ischemic brain eIF2alpha(P), and in conjunction with our previous report that eIF2alpha(P) is produced in the brain of reperfused PKR-/- mice, provides evidence that PERK is the kinase responsible for eIF2alpha phosphorylation in the early post-ischemic brain. PMID- 11389193 TI - Multiple oscillators in autonomic control. PMID- 11389194 TI - Functional and molecular expression of a voltage-dependent K(+) channel (Kv1.1) in interstitial cells of Cajal. AB - 1. Located within the gastrointestinal (GI) musculature are networks of cells known as interstitial cells of Cajal (ICC). ICC are associated with several functions including pacemaker activity that generates electrical slow waves and neurotransmission regulating GI motility. In this study we identified a voltage dependent K(+) channel (Kv1.1) expressed in ICC and neurons but not in smooth muscle cells. 2. Transcriptional analyses demonstrated that Kv1.1 was expressed in whole tissue but not in isolated smooth muscle cells. Immunohistochemical co localization of Kv1.1 with c-kit (a specific marker for ICC) and vimentin (a specific marker of neurons and ICC) indicated that Kv1.1-like immunoreactivity (Kv1.1-LI) was present in ICC and neurons of GI tissues of the dog, guinea-pig and mouse. Kv1.1-LI was not observed in smooth muscle cells of the circular and longitudinal muscle layers. 3. Kv1.1 was cloned from a canine colonic cDNA library and expressed in Xenopus oocytes. Pharmacological investigation of the electrophysiological properties of Kv1.1 demonstrated that the mamba snake toxin dendrotoxin-K (DTX-K) blocked the Kv1.1 outward current when expressed as a homotetrameric complex (EC(50) = 0.34 nM). Other Kv channels were insensitive to DTX-K. When Kv1.1 was expressed as a heterotetrameric complex with Kv1.5, block by DTX-K dominated, indicating that one or more subunits of Kv1.1 rendered the heterotetrameric channel sensitive to DTX-K. 4. In patch-clamp experiments on cultured murine fundus ICC, DTX-K blocked a component of the delayed rectifier outward current. The remaining, DTX-insensitive current (i.e. current in the presence of 10(-8) M DTX-K) was outwardly rectifying, rapidly activating, non inactivating during 500 ms step depolarizations, and could be blocked by both tetraethylammonium (TEA) and 4-aminopyridine (4-AP). 5. In conclusion, Kv1.1 is expressed by ICC of several species. DTX-K is a specific blocker of Kv1.1 and heterotetrameric channels containing Kv1.1. This information is useful as a means of identifying ICC and in studies of the role of delayed rectifier K(+) currents in ICC functions. PMID- 11389196 TI - Novel voltage-dependent non-selective cation conductance in murine colonic myocytes. AB - 1. Two components of voltage-gated, inward currents were observed from murine colonic myocytes. One component had properties of L-type Ca(2+) currents and was inhibited by nicardipine (5 x 10(-7) M). A second component did not 'run down' during dialysis and was resistant to nicardipine (up to 10(-6) M). The nicardipine-insensitive current was activated by small depolarizations above the holding potential and reversed near 0 mV. 2. This low-voltage-activated current (I(LVA)) was resolved with step depolarizations positive to -60 mV, and the current rapidly inactivated upon sustained depolarization. The voltage of half inactivation was -65 mV. Inactivation and activation time constants at -45 mV were 86 and 15 ms, respectively. The half-recovery time from inactivation was 98 ms at -45 mV. I(LVA) peaked at -40 mV and the current reversed at 0 mV. 3. I(LVA) was inhibited by Ni(2+) (IC(50) = 1.4 x 10(-5) M), mibefradil (10(-6) to 10(-5) M), and extracellular Ba(2+). Replacement of extracellular Na(+) with N-methyl-D glucamine inhibited I(LVA) and shifted the reversal potential to -7 mV. Increasing extracellular Ca(2+) (5 x 10(-3) M) increased the amplitude of I(LVA) and shifted the reversal potential to +22 mV. I(LVA) was also blocked by extracellular Cs(+) (10(-4) M) and Gd(3+) (10(-6) M). 4. Warming increased the rates of activation and deactivation without affecting the amplitude of the peak current. 5. We conclude that the second component of voltage-dependent inward current in murine colonic myocytes is not a 'T-type' Ca(2+) current but rather a novel, voltage-gated non-selective cation current. Activation of this current could be important in the recovery of membrane potential following inhibitory junction potentials in gastrointestinal smooth muscle or in mediating responses to agonists. PMID- 11389195 TI - Cyclic GMP regulation of the L-type Ca(2+) channel current in human atrial myocytes. AB - 1. The regulation of the L-type Ca(2+) current (I(Ca)) by intracellular cGMP was investigated in human atrial myocytes using the whole-cell patch-clamp technique. 2. Intracellular application of 0.5 microM cGMP produced a strong stimulation of basal I(Ca) (+64 +/- 5 %, n = 60), whereas a 10-fold higher cGMP concentration induced a 2-fold smaller increase (+36 +/- 8 %, n = 35). 3. The biphasic response of I(Ca) to cGMP was not mimicked by the cGMP-dependent protein kinase (PKG) activator 8-bromoguanosine 3',5' cyclic monophosphate (8-bromo-cGMP, 0.5 or 5 microM), and was not affected by the PKG inhibitor KT 5823 (100 nM). 4. In contrast, cGMP stimulation of I(Ca) was abolished by intracellular perfusion with PKI (10 microM), a selective inhibitor of the cAMP-dependent protein kinase (PKA). 5. Selective inhibition of the cGMP-inhibited phosphodiesterase (PDE3) by extracellular cilostamide (100 nM) strongly enhanced basal I(Ca) in control conditions (+78 +/- 13 %, n = 7) but had only a marginal effect in the presence of intracellular cGMP (+22 +/- 7 % in addition to 0.5 microM cGMP, n = 11; +20 +/ 22 % in addition to 5 microM cGMP, n = 7). 6. Application of erythro-9-[2 hydroxy-3-nonyl]adenine (EHNA, 30 microM), a selective inhibitor of the cGMP stimulated phosphodiesterase (PDE2), fully reversed the secondary inhibitory effect of 5 microM cGMP on I(Ca) (+99 +/- 16 % stimulation, n = 7). 7. Altogether, these data indicate that intracellular cGMP regulates basal I(Ca) in human atrial myocytes in a similar manner to NO donors. The effect of cGMP involves modulation of the cAMP level and PKA activity via opposite actions of the nucleotide on PDE2 and PDE3. PMID- 11389197 TI - Strong binding of myosin increases shortening velocity of rabbit skinned skeletal muscle fibres at low levels of Ca(2+). AB - 1. At low levels of activation, unloaded shortening of skinned skeletal muscle fibres takes place in two phases: an initial phase of high-velocity shortening followed by a phase of low-velocity shortening. The basis for Ca(2+) dependence of unloaded shortening velocity (V(o)) in the low-velocity phase was investigated by varying the level of thin filament activation with Ca(2+) and N-ethyl maleimide myosin subfragment-1 (NEM-S1), a non-tension-generating, strong binding derivative of subfragment-1. V(o) was measured with the slack-test method. 2. Treatment of skinned fibres with 5 microM NEM-S1 eliminated the low-velocity phase of shortening but had no effect on the high-velocity phase of shortening during submaximal activation with Ca(2+), or on V(o) during maximal activation with Ca(2+). 3. Extensive washout of NEM-S1 from the treated fibres restored the low-velocity phase of shortening and returned low-velocity V(o) to pre-treatment values. 4. The effect of NEM-S1 to increase low-velocity V(o) can be explained in terms of a model in which strong binding myosin cross-bridges activate the thin filament to a state in which the rate of ADP release from the actin-myosin-ADP complex and the rate of cross-bridge detachment from actin are accelerated during unloaded shortening. PMID- 11389198 TI - Excitation-contraction coupling in skeletal muscle of a mouse lacking the dihydropyridine receptor subunit gamma1. AB - 1. In skeletal muscle, dihydropyridine (DHP) receptors control both Ca(2+) entry (L-type current) and internal Ca(2+) release in a voltage-dependent manner. Here we investigated the question of whether elimination of the skeletal muscle specific DHP receptor subunit gamma1 affects excitation-contraction (E-C) coupling. We studied intracellular Ca(2+) release and force production in muscle preparations of a mouse deficient in the gamma1 subunit (gamma-/-). 2. The rate of internal Ca(2+) release at large depolarization (+20 mV) was determined in voltage-clamped primary-cultured myotubes derived from satellite cells of adult mice by analysing fura-2 fluorescence signals and estimating the concentration of free and bound Ca(2+). On average, gamma-/- cells showed an increase in release of about one-third of the control value and no alterations in the time course. 3. Voltage of half-maximal activation (V(1/2)) and voltage sensitivity (k) were not significantly different in gamma-/- myotubes, either for internal Ca(2+) release activation or for the simultaneously measured L-type Ca(2+) conductance. The same was true for maximal Ca(2+) inward current and conductance. 4. Contractions evoked by electrical stimuli were recorded in isolated extensor digitorum longus (EDL; fast, glycolytic) and soleus (slow, oxidative) muscles under normal conditions and during fatigue induced by repetitive tetanic stimulation. Neither time course nor amplitudes of twitches and tetani nor force-frequency relations showed significant alterations in the gamma1-deficient muscles. 5. In conclusion, the overall results show that the gamma1 subunit is not essential for voltage controlled Ca(2+) release and force production. PMID- 11389199 TI - Role of myoplasmic phosphate in contractile function of skeletal muscle: studies on creatine kinase-deficient mice. AB - 1. Increased myoplasmic inorganic phosphate (P(i)) has been suggested to have an important role in skeletal muscle fatigue, especially in the early phase. In the present study we used intact fast-twitch muscle cells from mice completely deficient in creatine kinase (CK(-/-)) to test this suggestion. These CK(-/-) muscle cells provide a good model since they display a higher P(i) concentration in the unfatigued state and fatigue without significant increase of P(i). 2. Tetanic contractions (350 ms duration) were produced in intact single muscle fibres. The free myoplasmic [Ca(2+)] ([Ca(2+)](i)) was measured with the fluorescent indicator indo-1. The force-[Ca(2+)](i) relationship was constructed from tetani at different frequencies. 3. Compared with wild-type fibres, CK(-/-) fibres displayed lower force in 100 Hz tetani and at saturating [Ca(2+)](i) (i.e. 100 Hz stimulation during caffeine exposure), higher tetanic [Ca(2+)](i) during the first 100 ms of tetanic stimulation, reduced myofibrillar Ca(2+) sensitivity when measurements were performed 100-200 ms into tetani, and slowed force relaxation that was due to altered cross-bridge kinetics rather than delayed Ca(2+) removal from the myoplasm. 4. In wild-type fibres, a series of 10 tetani resulted in reduced tetanic force, slowed force relaxation, and increased amplitude of [Ca(2+)](i) tails after tetani. None of these changes were observed in CK(-/-) fibres. 5. Complementary experiments on isolated fast-twitch extensor digitorum longus muscles showed a reduction of tetanic force and relaxation speed in CK(-/-) muscles similar to those observed in single fibres. 6. In conclusion, increased P(i) concentration can explain changes observed in the early phase of skeletal muscle fatigue. Increased P(i) appears to be involved in both fatigue induced changes of cross-bridge function and SR Ca(2+) handling. PMID- 11389200 TI - Endocytosis in identified rat corticotrophs. AB - 1. We used the patch-clamp technique, in conjunction with membrane capacitance measurement, fluorescence measurement of intracellular calcium concentration ([Ca(2+)](i)), and flash photolysis of caged Ca(2+) to study exo- and endocytosis in identified rat corticotrophs. 2. Exocytosis stimulated by depolarization pulses was typically followed by a 'slow' endocytosis that retrieved the membrane with a time constant of approximately 6 s. The efficiency (the endocytosis/exocytosis amplitude ratio) of 'slow' endocytosis was approximately 1.2 at [Ca(2+)](i) < 3 microM and increased to approximately 1.6 at [Ca(2+)](i) > 3 microM. 3. Whole-cell dialysis through a patch pipette did not affect the kinetics and the efficiency of 'slow' endocytosis, but the amplitude of exocytosis was reduced. 4. 'Slow' endocytosis did not require sustained [Ca(2+)](i) elevation and its kinetics was only weakly [Ca(2+)](i) dependent. Our results suggest that 'slow' endocytosis involves a Ca(2+) sensor with a high Ca(2+) affinity (approximately 500 nM). 5. At high [Ca(2+)](i) (> 10 microM), the 'slow' endocytosis was frequently preceded by a 'fast' endocytosis that comprised multiple steps of rapid decrease in membrane capacitance. 6. Neither calmodulin nor calcineurin appeared to be the Ca(2+) sensor for endocytosis because the two forms of endocytosis were not affected by the calmodulin inhibitor calmidazolium (500 microM) or the calcineurin inhibitors cyclosporin A (1 microM) and calcineurin autoinhibitory peptide (1 mg ml(-1)). Ba(2+), a poor activator of calmodulin, could support both forms of endocytosis but slowed the kinetics of 'slow' endocytosis approximately 2-fold. 7. Non-hydrolysable analogues of GTP (GDP-beta-S) and ATP (ATP-gamma-S) also failed to inhibit either form of endocytosis, indicating that neither GTP nor ATP was essential for endocytosis. 8. We suggest that the high Ca(2+) affinity of 'slow' endocytosis may be important for maintaining continuous cycles of exocytosis-endocytosis during sustained adrenocorticotropin secretion in corticotrophs. PMID- 11389201 TI - Calcium-calmodulin signalling pathway up-regulates glutamatergic synaptic function in non-pyramidal, fast spiking rat hippocampal CA1 neurons. AB - 1. The role of Ca(2+)-calmodulin (CaM) signalling cascades in modulating glutamatergic synaptic transmission on CA1 non-pyramidal fast-spiking neurons was investigated using whole-cell recording and perfusion in rat hippocampal slices. 2. Paired stimuli (PS), consisting of postsynaptic depolarization to 0 mV and presynaptic stimulation at 1 Hz for 30 s, enhanced excitatory postsynaptic currents (EPSCs) on non-pyramidal neurons in the stratum pyramidale (SP). The potentiation was reduced by the extracellular application of D-amino-5 phosphonovaleric acid (DAP-5, 40 microM), and blocked by the postsynaptic perfusion of 1,2-bis(2-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (BAPTA, 10 mM), a CaM-binding peptide (100 microM) or CaMKII (281-301) (an autoinhibitory peptide of CaM-dependent protein kinases, 100 microM). 3. The application of adenophostin, an agonist of inositol trisphosphate receptors (IP(3)Rs) that evokes Ca(2+) release, into SP non-pyramidal neurons via the patch pipette (1 microM) enhanced EPSCs and occluded PS-induced synaptic potentiation. The co application of BAPTA (10 mM) with adenophostin blocked synaptic potentiation. In addition, Ca(2+)-CaM (40:10 microM) induced synaptic potentiation, which occluded PS-induced potentiation and was attenuated by introducing CaMKII (281-301) (100 microM). EPSCs were sensitive to an antagonist of alpha-amino-3-hydroxy-5-methyl 4-isoxazole propionic acid receptor (AMPAR). 4. Application of Ca(2+)-CaM into SP non-pyramidal neurons induced the emergence of AMPAR-mediated EPSCs that were not evoked by low stimulus intensity before perfusion. Ca(2+)-CaM also increased the amplitude and frequency of spontaneous EPSCs. A scavenger of nitric oxide, carboxy-PTIO (30 microM in slice-perfusion solution), did not affect these increases in sEPSCs. 5. The magnitude of PS-, adenophostin- or Ca(2+)-CaM-induced synaptic potentiation in SP non-pyramidal neurons increased during postnatal development. 6. These results indicate that Ca(2+)-CaM signalling pathways in CA1 SP non-pyramidal neurons up-regulate glutamatergic synaptic transmission probably through the conversion of inactive-to-active synapses. PMID- 11389202 TI - Mechanisms underlying presynaptic facilitatory effect of cyclothiazide at the calyx of Held of juvenile rats. AB - 1. Excitatory postsynaptic currents (EPSCs) were recorded using the whole-cell patch-clamp technique at the calyx of Held synapse in the medial nucleus of the trapezoid body (MNTB) in auditory brainstem slices from juvenile rats. 2. Bath application of cyclothiazide (CTZ, 100 microM) significantly increased the amplitude of EPSCs mediated by alpha-amino-3-hydroxy-5-methyl-4 isoxazolepropionic acid (AMPA) and N-methyl-D-aspartate (NMDA) receptors. Cyclothiazide increased the magnitude of paired-pulse depression of both AMPA EPSCs (intervals, 50 and 500 ms) and NMDA-EPSCs (interval, 20 ms). In low Ca(2+), high Mg(2+) solution, CTZ decreased the number of failures and increased the mean amplitude of AMPA-EPSCs more than three-fold. 3. Presynaptic Ca(2+) currents and K(+) currents were directly recorded from the calyceal nerve terminals. These currents were attenuated by CTZ in a reversible manner. The magnitude of inhibition of presynaptic K(+) currents by CTZ (100 microM) was comparable to that by 5 microM 4-aminopyridine (4-AP). Both CTZ and 4-AP slowed the repolarizing phase of presynaptic action potentials. 4. The inhibitory effects of CTZ on presynaptic ion channels were mimicked by a solution having reduced Ca(2+) concentration and 5 microM 4-AP. This solution facilitated EPSCs, but the magnitude of facilitation was significantly less than that caused by CTZ. 5. In the presence of tetrodotoxin (TTX), CTZ increased the mean frequency of miniature EPSCs three-fold. CTZ prolonged their decay time but had no effect on their amplitude. The facilitatory effect of CTZ on the miniature frequency was neither blocked by a protein kinase C inhibitor nor occluded by phorbol ester, suggesting that a distinct mechanism underlies the effect of CTZ. 6. We conclude that CTZ facilitates transmitter release through suppression of presynaptic potassium conductance and stimulation of exocytotic machinery downstream of Ca(2+) influx. PMID- 11389203 TI - Ca(2+) signalling and PKCalpha activate increased endothelial permeability by disassembly of VE-cadherin junctions. AB - 1. The role of intracellular Ca(2+) mobilization in the mechanism of increased endothelial permeability was studied. Human umbilical vein endothelial cells (HUVECs) were exposed to thapsigargin or thrombin at concentrations that resulted in similar increases in intracellular Ca(2+) concentration ([Ca(2+)](i)). The rise in [Ca(2+)](i) in both cases was due to release of Ca(2+) from intracellular stores and influx of extracellular Ca(2+). 2. Both agents decreased endothelial cell monolayer electrical resistance (a measure of endothelial cell shape change) and increased transendothelial (125)I-albumin permeability. Thapsigargin induced activation of PKCalpha and discontinuities in VE-cadherin junctions without formation of actin stress fibres. Thrombin also induced PKCalpha activation and similar alterations in VE-cadherin junctions, but in association with actin stress fibre formation. 3. Thapsigargin failed to promote phosphorylation of the 20 kDa myosin light chain (MLC(20)), whereas thrombin induced MLC(20) phosphorylation consistent with formation of actin stress fibres. 4. Calphostin C pretreatment prevented the disruption of VE-cadherin junctions and the decrease in transendothelial electrical resistance caused by both agents. Thus, the increased [Ca(2+)](i) elicited by thapsigargin and thrombin may activate a calphostin C-sensitive PKC pathway that signals VE-cadherin junctional disassembly and increased endothelial permeability. 5. Results suggest a critical role for Ca(2+) signalling and activation of PKCalpha in mediating the disruption of VE-cadherin junctions, and thereby in the mechanism of increased endothelial permeability. PMID- 11389204 TI - Dendritic mechanisms underlying the coupling of the dendritic with the axonal action potential initiation zone of adult rat layer 5 pyramidal neurons. AB - 1. Double, triple and quadruple whole-cell voltage recordings were made simultaneously from different parts of the apical dendritic arbor and the soma of adult layer 5 (L5) pyramidal neurons. We investigated the membrane mechanisms that support the conduction of dendritic action potentials (APs) between the dendritic and axonal AP initiation zones and their influence on the subsequent AP pattern. 2. The duration of the current injection to the distal dendritic initiation zone controlled the degree of coupling with the axonal initiation zone and the AP pattern. 3. Two components of the distally evoked regenerative potential were pharmacologically distinguished: a rapidly rising peak potential that was TTX sensitive and a slowly rising plateau-like potential that was Cd(2+) and Ni(2+) sensitive and present only with longer-duration current injection. 4. The amplitude of the faster forward-propagating Na(+)-dependent component and the amplitude of the back-propagating AP fell into two classes (more distinctly in the forward-propagating case). Current injection into the dendrite altered propagation in both directions. 5. Somatic current injections that elicited single Na(+) APs evoked bursts of Na(+) APs when current was injected simultaneously into the proximal apical dendrite. The mechanism did not depend on dendritic Na(+)-Ca(2+) APs. 6. A three-compartment model of a L5 pyramidal neuron is proposed. It comprises the distal dendritic and axonal AP initiation zones and the proximal apical dendrite. Each compartment contributes to the initiation and to the pattern of AP discharge in a distinct manner. Input to the three main dendritic arbors (tuft dendrites, apical oblique dendrites and basal dendrites) has a dominant influence on only one of these compartments. Thus, the AP pattern of L5 pyramids reflects the laminar distribution of synaptic activity in a cortical column. PMID- 11389205 TI - Kinetic modification of the alpha(1I) subunit-mediated T-type Ca(2+) channel by a human neuronal Ca(2+) channel gamma subunit. AB - 1. Voltage-sensitive Ca(2+) channels (VSCCs) are often heteromultimeric complexes. The VSCC subtype specifically expressed by skeletal muscle has long been known to contain a gamma subunit, gamma(1), that is only expressed in this tissue. Recent work, initiated by the identification of the mutation present in the stargazer mouse, has led to the identification of a series of novel potential Ca(2+) channel gamma subunits expressed in the CNS. 2. Based on bioinformatic techniques we identified and cloned the human gamma(2), gamma(3) and gamma(4) subunits. 3. TaqMan analysis was used to quantitatively characterise the mRNA expression patterns of all the gamma subunits. All three subunits were extensively expressed in adult brain with overlapping but subunit-specific distributions. gamma(2) and gamma(3) were almost entirely restricted to the brain, but gamma(4) expression was seen in a broad range of peripheral tissues. 4. Using a myc epitope the gamma(2) subunit was tagged both intracellularly at the C-terminus and on a predicted extracellular site between the first and second transmembrane domains. The cellular distribution was then examined immunocytochemically, which indicated that a substantial proportion of the cellular pool of the gamma(2) subunit was present on the plasma membrane and provided initial evidence for the predicted transmembrane topology of the gamma subunits. 5. Using co-transfection techniques we investigated the functional effects of each of the gamma subunits on the biophysics of the T-type VSCC encoded by the alpha(1I) subunit. This revealed a substantially slowed rate of deactivation in the presence of gamma(2). In contrast, there was no significant corresponding effect of either gamma(3) or gamma(4) on alpha(1I) subunit-mediated currents. PMID- 11389206 TI - Dopamine D(2)-like receptors selectively block N-type Ca(2+) channels to reduce GABA release onto rat striatal cholinergic interneurones. AB - 1. The modulatory roles of dopamine (DA) in inhibitory transmission onto striatal large cholinergic interneurones were investigated in rat brain slices using patch clamp recording. 2. Pharmacologically isolated GABA(A) receptor-mediated IPSCs were recorded by focal stimulation within the striatum. Bath application of DA reversibly suppressed the amplitude of evoked IPSCs in a concentration-dependent manner (IC(50), 10.0 microM). 3. A D(2)-like receptor agonist, quinpirole (3-30 microM), also suppressed the IPSCs, whereas a D(1)-like receptor agonist, SKF 81297, did not affect IPSCs. Sulpiride, a D(2)-like receptor antagonist, blocked the DA-induced suppression of IPSCs (apparent dissociation constant (K(B)), 0.36 microM), while a D(1)-like receptor antagonist, SCH 23390 (10 microM), had no effect. 4. DA (30 microM) reduced the frequency of spontaneous miniature IPSCs (mIPSCs) without changing their amplitude distribution, suggesting that GABA release was inhibited, whereas the sensitivity of postsynaptic GABA(A) receptors was not affected. The effect of DA on the frequency of mIPSCs was diminished when extracellular Ca(2+) was replaced by Mg(2+) (5 mM), indicating that DA affected the Ca(2+) entry into the presynaptic terminal. 5. An N-type Ca(2+) channel selective blocker, omega-conotoxin GVIA (omega-CgTX, 3 microM), suppressed IPSCs by 65.4 %, whereas a P/Q-type Ca(2+) channel selective blocker, omega-agatoxin IVA (omega-Aga-IVA, 200 nM), suppressed IPSCs by 78.4 %. Simultaneous application of both blockers suppressed IPSCs by 95.9 %. Assuming a 3rd power relationship between Ca(2+) concentration and transmitter release, the contribution of N-, P/Q and other types of Ca(2+) channels to presynaptic Ca(2+) entry is estimated to be, respectively, 29.8, 40.0 and 34.5 % at this synapse. After the application of omega-CgTX, DA (30 microM) no longer affected IPSCs. In contrast, omega-Aga-IVA did not alter the level of suppression by DA, suggesting that the action of DA was selective for N-type Ca(2+) channels. 6. A G protein alkylating agent, N ethylmaleimide (NEM), significantly reduced the DA-induced suppression of IPSCs. 7. These results suggest that presynaptic D(2)-like receptors are present on the terminals of GABAergic afferents to striatal cholinergic interneurones, and down regulate GABA release by selectively blocking N-type Ca(2+) channels through NEM sensitive G proteins. PMID- 11389207 TI - Differences between nerve terminal impulses of polymodal nociceptors and cold sensory receptors of the guinea-pig cornea. AB - 1. Extracellular recording techniques were used to study nerve terminal impulses (NTIs) recorded from single polymodal nociceptors and cold-sensitive receptors in guinea-pig cornea isolated in vitro. 2. The amplitude and time course of NTIs recorded from polymodal nociceptors was different from those of cold-sensitive receptors. 3. Bath application of tetrodotoxin (1 microM) changed the time course of spontaneous NTIs recorded from both polymodal and cold-sensitive receptors. 4. Bath application of lignocaine (lidocaine; 1-5 mM) abolished all electrical activity. 5. Local application of lignocaine (2.5 and 20 mM) through the recording electrode changed the time course of the NTIs recorded from polymodal nociceptors but not that of NTIs recorded from cold-sensitive nerve endings. 6. It is concluded that action potentials propagate actively in the sensory nerve endings of polymodal nociceptors. In contrast, cold-sensitive receptor nerve endings appear to be passively invaded from a point more proximal in the axon where the action potential can fail or be initiated. PMID- 11389208 TI - Sequence dependence of post-tetanic potentiation after sequential heterosynaptic stimulation in the rat auditory cortex. AB - 1. To investigate the mechanisms for the coding stimulus sequence in the auditory cortex (AC), post-tetanic potentiation (PTP) was recorded after sequentially combined heterosynaptic stimulation was applied in rat AC slices. 2. Brief tetanic stimulation (TS) was applied at two sites on AC slices at intervals of 0.5-10 s. PTP of field potentials was induced by the earlier TS, rather than the later TS. PTP was followed by sequence-dependent long-term potentiation (LTP). 3. Using Ca(2+) imaging in the slices loaded with rhod-2, a Ca(2+) indicator, a sequence-dependent distribution of PTP was found in AC slices. 4. The sequence dependent PTP in excitatory postsynaptic potentials (EPSPs) was observed in supragranular pyramidal neurons. 5. The sequence dependence of PTP was not significantly affected by 1 microM bicuculline, an antagonist of GABA(A) receptors, or 100 microM 2-hydroxysaclofen, an antagonist of GABA(B) receptors. 6. Depolarization and firing recorded in pyramidal neurons during the later TS were less vigorous than when the slices were incubated in the control medium. However, this suppression of the responses during the later TS was not observed in the presence of 50 microM atropine, an antagonist of muscarinic receptors. 7. PTP was induced by the earlier and later TS in the presence of 50 microM atropine, so that the sequence dependence of PTP was abolished. Pirenzepine (50 microM), an antagonist of muscarinic M1 receptors, but not methoctramine (30 microM), an antagonist of M2 receptors, eliminated the sequence dependence of PTP. 8. These findings suggest that the sequence dependence of PTP in AC might have a role in the temporal processing of auditory information on the scale of seconds. PMID- 11389210 TI - Resetting of sympathetic rhythm by somatic afferents causes post-reflex coordination of sympathetic activity in rat. AB - 1. We have proposed previously that graded synchronous activity is produced by periodic inputs acting on weakly coupled or uncoupled oscillators influencing the discharges of a population of cutaneous vasoconstrictor sympathetic postganglionic neurones (PGNs) in anaesthetized rats. 2. Here we investigated the effects of somatic afferent (superficial radial nerve, RaN) stimulation, on the rhythmic discharges of this population. We recorded (1) at the population level from the ventral collector nerve and (2) from single PGNs focally from the caudal ventral artery of the tail. 3. Following RaN stimulation we observed an excitatory response followed by a period of reduced discharge and subsequent rhythmical discharges seemingly phase-locked to the stimulus. 4. We suggest that the rhythmical discharges following the initial excitatory response (conventional reflex) result from a resetting of sympathetic rhythm generators such that rhythmic PGN activity is synchronized transiently. We also demonstrate that a natural mechanical stimulus can produce a similar pattern of response. 5. Our results support the idea that in sympathetic control, resetting of multiple oscillators driving the rhythmic discharges of a population of PGNs may provide a mechanism for producing a sustained and coordinated response to somatic input. PMID- 11389209 TI - Spatial properties of koniocellular cells in the lateral geniculate nucleus of the marmoset Callithrix jacchus. AB - 1. The receptive field dimensions, contrast sensitivity and linearity of spatial summation of koniocellular (KC), parvocellular (PC) and magnocellular (MC) cells in the lateral geniculate nucleus (LGN) of 11 adult marmosets were measured using achromatic sinusoidal gratings. 2. The receptive field centre diameter of cells in each (PC, KC and MC) class increases with distance from the fovea. There is substantial overlap in centre size between the three cell classes at any eccentricity, but the PC cells have, on average, the smallest centres and the KC cells have the largest. Some PC and KC cells did not respond at all to the grating stimulus. 3. The contrast sensitivity of the receptive field centre mechanism in KC cells decreases in proportion to the centre area. A similar trend was seen for the surround mechanism. These characteristics are common to PC and MC cells, suggesting that they originate at an early stage of visual processing in the retina. 4. The KC cells showed, in general, lower peak evoked discharge rates than PC or MC cells. The spontaneous discharge rate of KC cells was lower than that of PC cells and similar to that of MC cells. 5. The majority of cells in all divisions of the LGN show linear spatial summation. A few cells did show non-linear spatial summation; these cells were predominantly located in the MC and ventral KC layers. 6. The ventral KC layers below and between the MC layers contain cells with larger and more transiently responding receptive fields than cells in the more dorsal KC layers. 7. We conclude that many of the contrast dependent spatial properties of cells in the marmoset LGN are common to PC, MC and KC cells. The main difference between KC cells and the other two classes is that there is more variability in their response properties, and they are less responsive to high spatial frequencies. PMID- 11389211 TI - Evidence for the role of alveolar epithelial gp60 in active transalveolar albumin transport in the rat lung. AB - 1. Transcytosis of albumin, involving the 60 kDa albumin-binding glycoprotein, gp60, was studied in cultured type II alveolar epithelial cells obtained from rat lungs. 2. Type II cells internalized the interfacial fluorescent dye RH 414, which marks for plasmalemma vesicles. Fluorescent forms of albumin and anti-gp60 antibody colocalized in the same plasmalemma vesicles. 3. Antibody (100 microg ml(-1)) cross-linking of gp60 for brief periods (15 min) markedly stimulated vesicular uptake of fluorescently tagged albumin. The caveolar disrupting agent, filipin (10 nM), abolished the stimulated internalization of albumin. 4. The vast majority of plasmalemmal vesicles carrying albumin also immunostained for caveolin-1; however, lysosomes did not stain for caveolin-1. Filipin depleted the epithelial cells of the caveolin-1-positive, albumin-transporting plasmalemma vesicles. 5. Prolonged (> 1 h) stimulation of type II cells with cross-linking anti-gp60 antibody produced loss of cell-surface gp60 and abolished endocytic albumin uptake. 6. Transalveolar transport of albumin was also studied in the isogravimetric rat lung preparation perfused at 37 degrees C. (125)I-labelled albumin was instilled into distal airspaces of lungs, and the resulting (125)I labelled albumin efflux into the vascular perfusate was determined. 7. Unlabelled albumin (studied over a range of 0-10 g (100 instilled ml)(-1)) inhibited 40 % of the transport of labelled albumin ((5.7 +/- 0.4) x 10(5) counts (instilled ml)( 1)) with an IC(50) value of 0.34 g (100 ml)(-1). 8. Filipin blocked the displacement-sensitive component of (125)I-labelled albumin transport, but had no effect on the transport of the paracellular tracer (3)[H]mannitol. 9. Displacement-sensitive (125)I-labelled albumin transport had a significantly greater Q(10) (27-37 degrees C) than the non-displaceable component. 10. Cross linking of gp60 by antibody instillation stimulated only the displacement sensitive (125)I-labelled albumin transalveolar transport in intact rat lungs. 11. To estimate the transport capacity of the displacement-sensitive system, the percentage of instilled (125)I-labelled albumin counts remaining in lung tissue was compared in lungs treated with instillates containing either 0.05 g (100 ml)( 1) unlabelled albumin or 5 g (100 ml)(-1) unlabelled albumin. Approximately 25 % of instilled (125)I-labelled albumin was cleared from the lung preparations per hour by the displacement-sensitive transport pathway. This component was blocked by filipin. PMID- 11389212 TI - Maternal undernutrition increases arterial blood pressure in the sheep fetus during late gestation. AB - 1. We have investigated the effect of a 50 % reduction in maternal nutrient intake during the last 30 days of pregnancy on arterial blood pressure and on arterial blood pressure responses to angiotensin II (AII) and the angiotensin converting enzyme (ACE) inhibitor captopril in the sheep fetus at 115-125 and at 135-145 days gestation (term = 147 +/- 3 days gestation). 2. Fetal plasma glucose concentrations were lower in the undernourished (UN) group compared to the control animals. There was no difference, however, in fetal plasma cortisol or adrenocorticotrophic hormone (ACTH) concentrations between the UN and control groups between 115 and 145 days gestation. 3. During the first 10 days of undernutrition, maternal plasma concentrations of cortisol were increased in the UN group compared to controls. At 115-125 days gestation, fetal arterial blood pressure was also higher in the UN group compared with controls and there was an inverse relationship (r = -0.62, P < 0.05) between mean arterial pressure and the fetal plasma concentrations of ACTH in the UN group. Fetal blood pressure responses to increasing doses of angiotensin II were also higher (P < 0.05) in UN compared to control animals at 115-125 days gestation. 4. Between 135 and 145 days gestation, fetal arterial blood pressure was increased in UN fetal sheep and mean arterial blood pressure was correlated with fetal plasma concentrations of cortisol. 5. Increased arterial blood pressure and responsiveness to AII measured in the fetuses of nutrient-restricted ewes may be related in part to fetal exposure to the actions of cortisol derived from transplacental transfer during the first 10 days after the start of the restricted feeding regime. PMID- 11389213 TI - Eccentric exercise-induced morphological changes in the membrane systems involved in excitation-contraction coupling in rat skeletal muscle. AB - 1. Physiological evidence suggests that excitation-contraction (E-C) coupling failure results from eccentric contraction-induced muscle injury because of structural and morphological damage to membrane systems directly associated with the E-C coupling processes within skeletal muscle fibres. In this study using rats, we observed the ultrastructural features of the membrane systems of fast twitch (FT) and slow-twitch (ST) muscle fibres involved in E-C coupling following level and downhill running exercise. Our aim was to find out whether mechanically mediated events following eccentric exercise caused disorder in the membrane systems involved in E-C coupling, and how soon after exercise such disorder occurred. We also compared the morphological changes of the membrane systems between ST and FT muscle fibres within the same muscles. 2. Single muscle fibres were dissected from triceps brachii muscles of male Fischer 344 rats after level or downhill (16 deg decline) motor-driven treadmill running (18 m min(-1), 5 min running with 2 min rest interval, 18 bouts). All single muscle fibres were histochemically classified into ST or FT fibres. The membrane systems were visualized using Ca(2+)-K(3)Fe(CN)(6)-OsO(4) techniques, and observed by high voltage electron microscopy (120-200 kV). 3. There were four obvious ultrastructural changes in the arrangement of the transverse (t)-tubules and the disposition of triads after the downhill running exercise: (1) an increase in the number of longitudinal segments of the t-tubule network, (2) changes in the direction and disposition of triads, (3) the appearance of caveolar clusters, and (4) the appearance of pentads and heptads (close apposition of two or three t tubule elements with three or four elements of terminal cisternae of the sarcoplasmic reticulum). The caveolar clusters appeared almost exclusively in the ST fibres immediately after downhill running exercise and again 16 h later. The pentads and heptads appeared almost exclusively in the FT fibres, and their numbers increased dramatically 2-3 days after the downhill running exercise. 4. The eccentric exercise led to the formation of abnormal membrane systems involved in E-C coupling processes. These systems have unique morphological features, which differ between ST and FT fibres, even within the same skeletal muscle, and the damage appears to be concentrated in the FT fibres. These observations also support the idea that eccentric exercise- induced E-C coupling failure is due to physical and chemical disruption of the membrane systems involved in the E-C coupling process in skeletal muscle. PMID- 11389214 TI - Carbohydrate ingestion attenuates the increase in plasma interleukin-6, but not skeletal muscle interleukin-6 mRNA, during exercise in humans. AB - 1. The present study was undertaken to examine the effects of exercise and carbohydrate (CHO) ingestion on interleukin-6 (IL-6) gene expression in skeletal muscle and plasma IL-6 concentration. 2. Seven moderately trained men completed 60 min of exercise at a workload corresponding to each individual's lactate threshold on four randomised occasions. Two trials were conducted on a bicycle ergometer (Cyc) and two on a running treadmill (Run) either with (CHO) or without (Con) the ingestion of a CHO beverage throughout the exercise. Muscle biopsies were obtained from the vastus lateralis before and immediately after exercise and IL-6 gene expression in these samples was determined using real-time PCR. In addition, venous blood samples were collected at rest, and after 30 min during and at the cessation of exercise. These samples were analysed for plasma IL-6. 3. Irrespective of exercise mode or CHO ingestion, exercise resulted in a 21 +/- 4 fold increase (P < 0.01; main exercise effect) in IL-6 mRNA expression. In contrast, while the mode of exercise did not affect the exercise-induced increase in plasma IL-6, CHO ingestion blunted (P < 0.01) this response. 4. These data demonstrate that CHO ingestion attenuates the plasma IL-6 concentration during both cycling and running exercise. However, because IL-6 mRNA expression was unaffected by CHO ingestion, it is likely that the ingestion of CHO during exercise attenuates IL-6 production by tissues other than skeletal muscle. PMID- 11389215 TI - Changes in passive tension of muscle in humans and animals after eccentric exercise. AB - 1. This is a report of experiments on ankle extensor muscles of human subjects and a parallel series on the medial gastrocnemius of the anaesthetised cat, investigating the origin of the rise in passive tension after a period of eccentric exercise. 2. Subjects exercised their triceps surae of one leg eccentrically by walking backwards on an inclined, forward-moving treadmill. Concentric exercise required walking forwards on a backwards-moving treadmill. For all subjects the other leg acted as a control. 3. Immediately after both eccentric and concentric exercise there was a significant drop in peak active torque, but only after eccentric exercise was this accompanied by a shift in optimum angle for torque generation and a rise in passive torque. In the eccentrically exercised group some swelling and soreness developed but not until 24 h post-exercise. 4. In the animal experiments the contracting muscle was stretched by 6 mm at 50 mm s(-1) over a length range symmetrical about the optimum length for tension generation. Measurements of passive tension were made before and after the eccentric contractions, using small stretches to a range of muscle lengths, or with large stretches covering the full physiological range. 5. After 150 eccentric contractions, passive tension was significantly elevated over most of the range of lengths. Measurements of work absorption during stretch release cycles showed significant increases after the contractions. 6. It is suggested that the rise in passive tension in both human and animal muscles after eccentric contractions is the result of development of injury contractures in damaged muscle fibres. PMID- 11389216 TI - Muscle and joint elastic properties during elbow flexion in Duchenne muscular dystrophy. AB - 1. Maximal voluntary contraction (MVC), series elastic stiffness and total joint stiffness during elbow flexion were investigated in healthy boys and in boys with Duchenne muscular dystrophy (DMD) in order to assess changes in mechanical properties induced by the disease. 2. Two methods were used to perform stiffness measurements: (i) the application of sinusoidal perturbations to the joint during flexion efforts, allowing the calculation of total joint stiffness; (ii) the use of quick-release movements of the elbow, which had previously been maintained in isometric contraction, allowing the calculation of series elastic stiffness. In each case, stiffness was linearly related to torque, leading to the calculation of a normalized stiffness index as the slope of this stiffness-torque relationship. 3. As expected, mean MVC was found to be much higher for healthy boys (20.02 +/- 5.20 N m) than for DMD patients (3.09 +/- 2.44 N m). Furthermore, the results showed that it was possible to characterize healthy and DMD children by virtue of the mechanical properties measured. Mean series elastic stiffness index was higher for DMD children (142.55 +/- 136.58 rad(-1)) than for healthy subjects (4.39 +/- 2.53 rad(-1)). The same holds for mean total joint stiffness index: 43.68 +/- 67.58 rad(-1) for DMD children and 2.26 +/- 0.70 rad(-1) for healthy subjects. In addition, increases in stiffness were more marked in DMD patients exhibiting high levels of muscle weakness. 4. These changes are interpreted in terms of the adaptation of the properties of the muscles and joint involved, i.e. muscle fibres, tendons, peri- and intra-articular structures. PMID- 11389217 TI - Interlimb co-ordination in human infant stepping. AB - 1. We held infants (aged 4-12 months) over a treadmill to study how they co ordinated the two limbs during stepping. We disturbed one limb during the stance or swing phase and recorded the responses (muscle activity and movement) from both lower limbs. Manual disturbances were applied during the stance phase by sliding the foot backward, forcing the limb into the swing phase. Disturbances were also applied in the swing phase by manually extending the hip, interfering with the forward motion of the limb. Additional disturbances were applied to see if both limbs could perform the stance and swing phase synchronously. 2. When the limb was forced to initiate the swing phase on one side, the contralateral limb either prolonged its contact with the ground or quickly established ground contact. When the forward motion of the limb was interrupted in the swing phase, the swing phase was prolonged on the disturbed side and the stance phase prolonged on the contralateral side. In most cases, one leg maintained ground contact. Moreover, it was easy to elicit bilateral, simultaneous stance phase, whereas it was difficult to elicit simultaneous swing phase. In cases where swing phase in the two limbs was initiated close in time, rhythmic alternate stepping was immediately restored in the following step. 3. We conclude that human infants can generate co-ordinated motor responses bilaterally in response to unilateral perturbations, well before the onset of independent walking. PMID- 11389218 TI - Fibrinogen: structure, function, and surface interactions. AB - Fibrinogen plays a central role in the mechanism of coagulation and thrombosis and is partially involved in the development of postintervention restenosis. Because of therapeutic implications, it is convenient for the vascular interventionalist to revisit its structure, function, and relationships within the vascular environment. This review focuses on the molecular structure, mechanisms of polymerization and lysis, and fibrinogen interaction with the platelet alpha(IIb)beta(3) [corrected] integrin. It also addresses the less understood interaction of fibrinogen with artificial surfaces. Glycoprotein IIb IIIa blockers, targeted to interfere with fibrinogen-platelet interactions, widely used in clinical practice, are discussed, and trials of new drugs are also summarized. PMID- 11389219 TI - Transcatheter interventions for the treatment of peripheral atherosclerotic lesions: part I. AB - Transcatheter endovascular procedures are increasingly used to treat symptomatic peripheral atherosclerosis. This two-part review identifies the existing evidence supportive of the application of transcatheter treatments for peripheral atherosclerotic lesions. The first part addresses the treatment of obstructive lesions that cause limb claudication and critical ischemia, renovascular hypertension and azotemia, and mesenteric ischemia. Studies were identified via a search of MEDLINE (January 1993 through April 1999) and reference lists of identified articles. When multicenter prospective randomized trials or other high quality studies were unavailable, a preference was given to studies with at least 50 patients per treated group and a minimum mean follow-up duration of 6 months. Data presented in tables are proportionally weighted averages from included studies. For each application, the authors assessed the quality of evidence (QOE; efficacy, safety, and, where available, cost-effectiveness) and made recommendations with appropriate caveats. There is higher QOE supporting the more established treatments such as lower limb percutaneous transluminal angioplasty (PTA) with stent placement and thrombolysis. Treatments such as renal artery PTA and stent placement and mesenteric and brachiocephalic PTA are in wide use, but high QOE supporting general application is lacking. Blanket recommendations based on established efficacy and cost-effectiveness cannot be made. However, the use of transcatheter therapies can be supported in specific circumstances based on an expected reduction in procedure-related morbidity and/or mortality rates. It is hoped that the identification of deficiencies in the literature will inform and inspire critically needed research in this area. PMID- 11389220 TI - Recanalization of the obstructed nasolacrimal duct system. PMID- 11389221 TI - Nasolacrimal stents in the management of epiphora: medium-term results of a multicenter prospective study. AB - PURPOSE: To evaluate in a prospective multicenter setting the clinical utility of polyurethane stents in the percutaneous management of epiphora. MATERIALS AND METHODS: Patients (N = 163; age range = 22-85 y, mean = 52 y; 29 men, 134 women) with severe epiphora had stents (n = 183) inserted under fluoroscopic guidance in 180 lacrimal systems (unilateral = 146; bilateral = 17) to treat complete (n = 172) or partial (n = 8) obstruction of the nasolacrimal duct or sac. The junction between sac and duct was the most frequent location (n = 102), followed by the sac alone (n = 48), and the duct alone (n = 30). The etiology of the obstruction was idiopathic in 113 cases (63%) and chronic dacryocystitis in 67 (37%). The set designed by Song was used in all patients and the original technique was slightly modified by the authors. All patients were treated on an outpatient basis. Average time of the procedure was 14 minutes (range = 3-70 min). RESULTS: Initial technical success rate of stent placement was 97%. Resolution of epiphora was complete in 175 eyes and partial in five. On follow-up (mean = 450 d; range = 8 730 d), 157 of 183 stents remained patent (85.8%). Of the 24 obstructed, 19 were easily withdrawn and 17 of these patients remained asymptomatic for a mean of 15 months (secondary patency rate = 89.5%). CONCLUSIONS: The procedure is simple and safe. It can be performed on an outpatient basis and the original technique could be improved with some technical modifications. It is well tolerated by patients and may be considered as a valid alternative technique for the resolution of epiphora. PMID- 11389222 TI - Treatment of hemodialysis catheter-associated fibrin sheaths by rt-PA infusion: critical analysis of 124 procedures. AB - PURPOSE: To prospectively evaluate the efficacy of a low-dose, 3-hour infusion of recombinant tissue plasminogen activator (rt-PA) for the treatment of hemodialysis catheter (HDC)-associated fibrin sheaths. This report expands the authors' experience with this technique over that previously reported. MATERIALS AND METHODS: Fifty-five patients with end-stage renal disease (38 women, 17 men) undergoing catheter-directed hemodialysis treatment were evaluated for 124 episodes of HDC dysfunction. This patient group had a mean age of 57 years and an age range of 23-92 years. Radiographic contrast studies and/or clinical evaluation were consistent with the presence of a fibrin sheath on the arterial and/or venous port in all cases. Each patient underwent a thrombolytic infusion consisting of 2.5 mg rt-PA in 50 mL normal saline at 17 mL/h (3-hour infusion) per port. All infusions were performed in the interventional radiology recovery room on an outpatient basis. Patients were followed prospectively for technical success, complications, catheter patency, and long-term outcome. RESULTS: The technical success rate, defined as return of effortless manual aspiration and infusion capability from both ports followed by at least one successful dialysis session, was 91%. No patient was excluded from rt-PA therapy because of contraindications, and the procedure-related complication rate was zero percent. A Kaplan-Meier survival analysis yielded primary patency rates at 30, 60, 90, and 120 days of 0.55, 0.36, 0.25, and 0.15 (SE <.10), respectively; secondary patency rates at 60, 120, 180, and 240 days were 0.70, 0.46, 0.30, and 0.27 (SE <.10), respectively (P < 001). At the end of the study period, all 52 surviving patients continued to undergo catheter-directed hemodialysis and 34 (65%) were using the same catheter present at the time of entrance into the study. Of the 18 patients (35%) requiring catheter exchange, 16 (89%) did for persistent malfunction after rt-PA therapy, one (5.5%) for infection, and one (5.5%) for a fractured hub. CONCLUSION: Thrombolytic therapy with use of a 2.5-mg rt-PA infusion through each port over a 3-hour period would appear to be a safe method for treating HDC associated fibrin sheaths. Immediate return of catheter function is achieved in most patients, obviating more invasive techniques. Primary patency rates are relatively short, but catheters that fail can be retreated, resulting in secondary patency rates that are substantial and significantly improved. PMID- 11389223 TI - Osteoid osteoma: CT-guided percutaneous radiofrequency ablation and follow-up in 47 patients. AB - PURPOSE: To evaluate computed tomography (CT)-guided radiofrequency (RF) ablation as a minimally invasive therapy for osteoid osteoma with regard to technical and clinical success and immediate and delayed complications. MATERIALS AND METHODS: Forty-seven patients (age range, 8-41 y; mean age, 19.6 y) with osteoid osteomas (femur, n = 25; tibia, n = 15; pelvis, n = 2; humerus, n = 1; ulna, n = 1; talus, n = 1; calcaneus, n = 1; vertebral body, n = 1) were treated with CT-guided RF ablation in 15 cases after one (n = 10) or two (n = 5) unsuccessful attempts at open surgical resection. Percutaneous therapy was performed with use of general or spinal anesthesia. After localization of the nidus with 1-3-mm CT sections, osseous access was established with either a 2-mm coaxial drill system or an 11 gauge Jamshidi needle. RF ablation was performed at 90 degrees C for a period of 4-5 minutes with use of a rigid RF electrode with a diameter of 1 mm. The procedures were regarded as technically successful if the tip of the RF electrode could be placed within the center of the nidus and could be heated to the desired temperature. Clinical success of treatment was defined as permanent relief of pain and return to normal function without additional treatment. In case of persistence or recurrence of symptoms after RF ablation, treatment was regarded as secondarily successful if permanent relief of symptoms could be achieved in a second procedure. RESULTS: All procedures were technically successful. Clinical success was achieved in 94% of patients (44 of 47). Three patients had recurrence of pain 3, 5, and 7 months after treatment, respectively (mean observation interval, 22 mo). All recurrences were treated successfully in a second procedure (secondary success rate, 100%). No immediate or delayed complications were observed. CONCLUSION: CT-guided percutaneous RF ablation is a simple, minimally invasive, safe and highly effective technique for treatment of osteoid osteoma. PMID- 11389224 TI - Peripherally inserted central catheters: outcome as a function of the operator. AB - PURPOSE: To determine the natural history of and outcome involved with peripherally inserted central catheters (PICCs) placed at a single institution and examine potential differences in the natural history of PICCs placed by interventional radiologists (IRs) versus registered nurses (RNs). MATERIALS AND METHODS: A prospective analysis of all patients receiving PICCs at one academic medical center over a period of 6.5 months was conducted. At our institution, PICCs are placed primarily by RN members of the intravenous team. Placement procedures deemed unfeasible or problematic by RNs are referred to an IR for insertion under fluoroscopic guidance. A total of 322 PICCs (130 by IRs, 192 by RNs) were successfully placed in 256 patients. In three patients, placement was attempted but was a technical failure by both RNs and IRS: Seven patients in each group were lost to follow-up. PICCs were classified as successfully completed therapy or as having been prematurely removed, which was further stratified into suspected infection, occlusion, phlebitis, mechanical failure, inadvertent patient removal, and other. RESULTS: Overall rate of premature removal for PICCs placed by IRs versus RNs was not significantly different (30.8% vs 23.4%, respectively). PICCs placed by IRs had an increased rate of occlusion (IRs = 9.2%, RNs = 3.6%; P =.02). Other reasons for premature removal did not differ in incidence. Overall, PICCs were successfully placed in 99.1% of all patients and the course of therapy was completed in 69.3%. CONCLUSION: It is reasonable and cost-effective for trained RNs to place PICCs whenever feasible and refer complicated placements to IRS: PMID- 11389226 TI - Evaluation of the LGM Vena-Tech infrarenal vena cava filter in an ovine venous thromboembolism model. AB - PURPOSE: To validate a new percutaneous model of venous thrombosis in sheep and evaluate the use of the LGM Vena-Tech vena cava filter with use of this model. MATERIALS AND METHODS: After implantation of a LGM Vena-Tech filter in the infrarenal vena cava (IVC), thrombus was obtained by blocking the iliac vein with an inflated balloon (Wedge catheter) for 15 minutes and simultaneously injecting 20 mL of fresh thrombus into the femoral vein. Clot migration of the thrombus was induced by balloon deflation and injection of contrast medium. Migration and capture of the thrombus by the filter were filmed under fluoroscopy at 1 frame/sec. Euthanasia followed by pathologic examination of the IVC, heart, and lungs was performed immediately after the procedure in five sheep (group 1). Sheep in groups 2, 3, and 4, (five in each group), were killed at 2, 4, and 8 weeks, respectively, after vena cavography. Histologic examination was performed to analyze the evolution of the thrombus captured, the incorporation of the filter in the caval wall, and the physical and mechanical effects of captured thrombi on the filter. RESULTS: The Vena-Tech filter captured a large amount of thrombus in all cases except one, in which the filter captured a small strand of thrombus, related to incomplete occlusion of the iliac vein during the clot formation procedure. In the 15 animals in which follow-up was performed, fibrous evolution of the thrombus was observed at gross examination in 14, leading to the formation of fibrous webs between the filter and the IVC wall in eight. These observations were not related to the incidence of filter retraction (n = 4), caudal migration (n = 2), tilt (n = 2), or caval perforation (n = 1). Heart and lung thrombi were present in six animals. Histologic results confirmed the fibrous evolution of the thrombus and its organization during follow-up. Neointima increases significantly (P <.02) during follow-up, from 135.7 microm +/ 13.4 at 2 weeks to 192.2 microm +/- 125.7 at 4 weeks and 334.2 microm +/- 144.1 at 8 weeks. CONCLUSION: The model used is suitable for the formation of a large amount of venous thrombus and analysis of its migration and capture by the LGM Vena-Tech filter. Fibrous evolution of the thrombus, including development of webs and changes in filter shape and position, were the main outcomes observed. PMID- 11389225 TI - Isolated lower extremity chemotherapeutic infusion for treatment of osteosarcoma: experimental study and preliminary clinical report. AB - PURPOSE: To increase the dose of drug delivered to a tumor while maintaining tolerable systemic side effects, an interventional technique of isolated lower extremity infusion was investigated. MATERIALS AND METHODS: Experiments were performed in eight dogs. Four dogs were treated by a combination of intraarterial (IA) femoral cisplatin infusion at a dose of 2.0 mg/kg with drug removal from the ipsilateral extremity venous blood by a dialyzer. The other four dogs comprised the control group. In these animals, left femoral arterial IA cisplatin infusion was performed without dialysis. Leukocyte and platelet counts, blood urea nitrogen (BUN) levels, and serum creatinine levels were recorded before and after the treatment. Subsequently, two human patients with inoperable osteosarcoma were treated with the isolated infusion. RESULTS: In the experiments, 85%-90% of the free platinum that entered the dialyzer was removed. The peak systemic plasma cisplatin concentrations in animals undergoing dialysis were reduced by 81.25% compared to those in animals undergoing femoral IA infusion without hemodialysis. There were no significant changes in the hematologic profiles or BUN and serum creatinine levels in the experimental animals. However, in the control group, all dogs developed myelosuppression and severe renal toxicity after IA infusion of the same dose of cisplatin. Clinically, immediate relief of symptoms related to the primary tumor was achieved in both human patients after the combination of isolated IA infusion and embolotherapy. CONCLUSION: Single-pass hemodialysis removed a significant amount of cisplatin after regional IA infusion, reduced systemic toxicity, and permitted survival of the experimental animals. In two patients with osteosarcoma, percutaneous isolated lower extremity chemotherapeutic infusion therapy and embolotherapy were performed safely with partial responses. PMID- 11389227 TI - Evaluation of renal arteries with use of gadoterate meglumine-, CO(2)-, and iodixanol-enhanced DSA measurements versus histomorphometry in renal artery restenosis in rabbits. AB - PURPOSE: To experimentally evaluate gadolinium (Gd)-, carbon dioxide (CO(2))-, and iodixanol-enhanced digital subtraction angiography (DSA) versus histomorphometry in the assessment of renal artery stenosis. MATERIALS AND METHODS: Fifteen male New Zealand White rabbits weighing 4.0 kg underwent percutaneous catheterization. Renal artery stenosis was induced by bilateral overdilation-deendothelialization (balloon diameter = 2 mm). The percentage of artery overdilation was 33%. After 4 weeks, the rabbits were randomized into two groups: group A underwent right-sided therapeutic percutaneous transluminal renal angioplasty (PTRA) (balloon diameter = 1.5 mm). After another 4 weeks, the renal arteries were evaluated by gadoterate-, iodixanol-, and CO(2)-enhanced selective quantitative DSA. The rabbits were then killed and renal arteries were perfusion fixed for 60 minutes. Serial orcein-stained 4-um-thick slices were prepared for histomorphometry. RESULTS: Based on morphometric data of single-stenosis versus post-PTRA restenosis lesions, no significant difference was observed between Gd- and iodixanol-enhanced quantitative DSA (r(2) > 0.95), although the iodine/Gd density ratio was equal to 3.5. Carbon dioxide less reliably allowed quantitative DSA (r(2) < 0.75). CONCLUSION: Gd-based contrast agents represent a highly reliable alternative in experimental quantitative DSA evaluation of renal artery restenosis. PMID- 11389228 TI - Real-time MR Guidance for inferior vena cava filter placement in an animal model. AB - It was the aim of this study to examine the feasibility of real-time magnetic resonance (MR) imaging for MR-guided placement of inferior vena cava (IVC) filters, which were placed in five pigs via a femoral approach. The introducer sheath and dilator were marked with Dysprosium rings. The procedures were performed under MR guidance with use of a 1.5-T ACS-NT imager. Radial filling of k-space in conjunction with the sliding window reconstruction technique achieved real-time MR imaging with a frame rate of 20 images/sec. Simultaneous real-time visualization of the vascular anatomy and interventional instruments was achieved under real-time conditions and allowed correct placement of IVC filters in all five cases as confirmed by radiographic angiography. PMID- 11389229 TI - Treatment of fibromuscular dysplasia and renal artery aneurysm with use of a stent-graft. AB - A Jostent stent-graft was placed in a renal artery with a focal stenosis and an associated aneurysm in a young patient with fibromuscular dysplasia. Arteriographic follow-up at 16 months showed that the stent-graft remained patent and the aneurysm was excluded. PMID- 11389230 TI - Neurologic injury after endovascular stent-graft and bilateral internal iliac artery embolization for infrarenal abdominal aortic aneurysm. AB - The authors report a rare neurologic complication after the implantation of a bifurcated stent-graft for abdominal aortic aneurysm. The stent-graft was extended to both external iliac arteries after embolization of both internal iliac arteries. The patient subsequently had weakness and numbness of both lower limbs with bowel and bladder incontinence. He probably had ischemic injury to the nerve roots or the lumbosacral plexus, which was related to extensive occlusion of their supplying arteries. The mechanism of spinal cord and neurologic ischemia after aortic stent-graft implantation is discussed. PMID- 11389231 TI - Evaluation of various maneuvers for prevention of air embolism during central venous catheter placement. AB - This study is designed to evaluate the various physiologic maneuvers (Valsalva, humming, breath-hold) for the potential prevention of air embolism during central venous catheter placement. Central venous pressure measurements were prospectively obtained in 40 patients undergoing central venous catheter placement. The average central venous pressure at baseline was 3.275 mm Hg (range = -4 to 16, SD = 5.99). The average central venous pressure during breath hold was 6.1 mm Hg (range = -6 to 24, SD = 7.99). The average central venous pressure during humming was 5.1 mm Hg (range = -4 to 20, SD = 6.4) The average central venous pressure during the Valsalva maneuver was 18.43 (range = -3 to 48, SD = 14.73). Forty percent of patients (16 of 40) had negative central venous pressures at rest, 25% (10 of 40) had negative pressures during breath hold, 20% (8 of 40) had negative pressures during humming, and 2.5% (1 of 40) had negative pressures during Valsalva maneuver. The average increases in central venous pressure during breath hold, humming, and Valsalva were 2.85, 1.82, and 15.2 mm Hg, respectively. The difference between pressures during Valsalva and other maneuvers was statistically significant (P <.05). The conclusion is that the Valsalva maneuver is superior to breath-hold and humming for increasing central venous pressure during central venous catheter placement and, therefore, it is more likely to prevent air embolism in cooperative patients. PMID- 11389232 TI - Transcatheter occlusion of a large pulmonary arteriovenous malformation with use of a Cardioseal device. AB - Large pulmonary arteriovenous malformations (PAVMs) carry a significant risk of neurologic complications and present technical difficulties in transcatheter treatment with use of coils or detachable balloons. A 26-year-old man with a giant PAVM, who had undergone unsuccessful attempted closure with use of a Gianturco-Grifka occlusion device in the past, underwent successful transcatheter embolization with two Cardioseal double umbrella devices designed for occlusion of intracardiac communications. The procedure was technically easy, had no complications, and provided sustained improvement in arterial saturation and exercise tolerance during follow-up. Transcatheter double umbrella device occlusion of large arteriovenous malformations is feasible and should be considered, especially for very large fistulas. PMID- 11389233 TI - Fatal paradoxic embolism occurring during IVC filter insertion in a patient with chronic pulmonary thromboembolic disease. AB - A patient diagnosed with chronic thromboembolic disease experienced a fatal intraprocedural paradoxic embolism during inferior vena cava (IVC) filter insertion. The frequency of patent intra-atrial shunts in patients with chronic thromboembolic disease is surprisingly high, occurring in approximately 29.5% of patients with chronic thromboembolic disease. Prevention of embolic events during venous interventions in such cases requires that the operator recognize that central shunts may be present and strictly adhere to meticulous technique during IVC filter insertion. PMID- 11389234 TI - A hemorrhagic complication of neurofibromatosis. PMID- 11389235 TI - Common bronchial artery trunk originating from the left subclavian artery. PMID- 11389236 TI - Re: the Legs For Life screening for peripheral vascular disease: compliance with physician recommendations in moderate- and high-risk assessed patients. PMID- 11389237 TI - Fever phobia revisited: have parental misconceptions about fever changed in 20 years? AB - OBJECTIVES: Fever is one of the most common reasons that parents seek medical attention for their children. Parental concerns arise in part because of the belief that fever is a disease rather than a symptom or sign of illness. Twenty years ago, Barton Schmitt, MD, found that parents had numerous misconceptions about fever. These unrealistic concerns were termed "fever phobia." More recent concerns for occult bacteremia in febrile children have led to more aggressive laboratory testing and treatment. Our objectives for this study were to explore current parental attitudes toward fever, to compare these attitudes with those described by Schmitt in 1980, and to determine whether recent, more aggressive laboratory testing and presumptive treatment for occult bacteremia is associated with increased parental concern regarding fever. METHODS: Between June and September 1999, a single research assistant administered a cross-sectional 29 item questionnaire to caregivers whose children were enrolled in 2 urban hospital based pediatric clinics in Baltimore, Maryland. The questionnaire was administered before either health maintenance or acute care visits at both sites. Portions of the questionnaire were modeled after Schmitt's and elicited information about definition of fever, concerns about fever, and fever management. Additional information included home fever reduction techniques, frequency of temperature monitoring, and parental recall of past laboratory workup and treatment that these children had received during health care visits for fever. RESULTS: A total of 340 caregivers were interviewed. Fifty-six percent of caregivers were very worried about the potential harm of fever in their children, 44% considered a temperature of 38.9 degrees C (102 degrees F) to be a "high" fever, and 7% thought that a temperature could rise to >/=43.4 degrees C (>/=110 degrees F) if left untreated. Ninety-one percent of caregivers believed that a fever could cause harmful effects; 21% listed brain damage, and 14% listed death. Strikingly, 52% of caregivers said that they would check their child's temperature 30 days in 13.2% of the episodes. Six of the 173 children (3.5%) developed on 1 or more occasions tube otorrhea that failed to improve satisfactorily with conventional outpatient management. Five of these children were hospitalized to receive parenteral antibiotic treatment, 1 child twice and 1 three times, and 1 also underwent tube removal. The sixth child underwent tube removal as an outpatient. CONCLUSIONS: Tube otorrhea is a common and often recurrent and/or stubborn problem in young children who have undergone tube placement for persistent MEE. The extent of the problem seems to be related inversely to socioeconomic status. Tube otorrhea does not always respond satisfactorily to outpatient management and for resolution may require parenteral antimicrobial treatment and/or tube removal. PMID- 11389240 TI - Mortality risk in children with epilepsy: the Dutch study of epilepsy in childhood. AB - OBJECTIVE: Long-term follow-up studies of patients with epilepsy have revealed an increased mortality risk compared with the general population. Mortality of children who have epilepsy in modern times is as yet unknown. Therefore, the objective of this study was to determine mortality of children who have epilepsy in comparison with the general population. METHODS: Between August 1988 and August 1992, 472 children, aged 1 month to 16 years, who presented in 1 of the participating hospitals with 2 or more newly diagnosed unprovoked seizures or at least 1 status epilepticus were enrolled in the study. All children were followed for 5 years or until death. The number of deaths observed during follow-up was compared with the expected number of deaths in the same age group in the general population in the Netherlands. RESULTS: Nine children died during follow-up, amounting to a mortality rate of 3.8/1000 person-years, which is sevenfold higher than expected (95% confidence interval = 2.4-11.5). No deaths were observed among the 328 children who had epilepsy of nonsymptomatic cause. All deceased children had epilepsy that was caused by a static or progressive neurologic disorder (mortality risk = 22.9; 95% confidence interval = 7.9-37.9). None of them died from sudden unexpected and unexplained death of epilepsy. CONCLUSIONS: In our cohort, we found no indication that children who have nonsymptomatic epilepsy have an increased mortality risk compared with the general population, whereas children who have symptomatic epilepsy have a 20-fold increased mortality risk. These data provide guidance for counseling parents of children who have epilepsy. PMID- 11389241 TI - Transcutaneous bilirubin measurement: a multicenter evaluation of a new device. AB - OBJECTIVES: The early discharge of neonates from hospitals makes transcutaneous measurement of total bilirubin concentration a useful tool to monitor neonatal jaundice. The objectives of this study were to determine whether 1) transcutaneous bilirubin (TcB) measurement, as performed using BiliCheck (BC), correlates with total serum bilirubin (TSB) levels, measured with standard laboratory methods and with high-pressure liquid chromatography (HPLC-B); 2) infant race, gestational age, postnatal age, or body weight interferes with the measurement of TcB levels in newborn infants; 3) the variability of the TcB measurement is comparable to the variability of TSB measurements; and 4) TcB measurements obtained from the forehead (BCF) and sternum (BCS) generate comparable results. STUDY DESIGN: Newborn infants who were <28 days and >30 weeks' gestational age and who underwent tests for TSB as part of their normal care in 6 different European hospitals were studied. A total of 210 infants were enrolled in the study, 35 at each site. Near simultaneous (within +/- 30 minutes) blood collection for TSB and BCF and BCS measurements were performed. TSB levels were determined by the serum bilirubin method in use at each site, and all HPLC-B determinations were made at the same, independent laboratory. RESULTS: The study group consisted of 140 white, 31 Asian, 14 Hispanic, 9 African, and another 16 newborns of different races. The correlation coefficient (r) between BCF and HPLC B was 0.890 (95% confidence interval = 0.858-0.915). BCF and BCS generated similar results (r value = 0.890 for BCF and 0.881 for BCS), even if BCS slightly overestimated (mean error = -0.04 mg/dL) and BCF slightly underestimated (mean error = 0.96 mg/dL) in comparison with HPLC-B. Analysis of covariance demonstrated that BC accuracy was independent of race, birth weight, gestational age, and postnatal age of the newborn. Receiver operating characteristic curves were evaluated for BCF and TSB, each compared with HPLC-B. With the use of a cutoff point for HPLC-B of 13 mg/dL (222 micromol/L) and a cutoff of 11 mg/dL on the BCF and TSB, similar sensitivity/specificity (93%/73% for BCF, 95%/76% for TSB) were observed. The use of a cutoff point for HPLC-B of 17 mg/dL (290 micromol/L) and 14 mg/dL (240 micromol/L) for BCF and TSB also produced similar sensitivity/specificity (90%/87% for the BC and 87%/83% for TSB). CONCLUSIONS: Because the correlation coefficient for HPLC-B and BCF is very similar to that found for HPLC-B and laboratory TSB, BC could be used not only as a screening device but also as a reliable substitute of TSB determination. At higher levels of TSB, in which phototherapy and/or exchange transfusion might be considered, BC performed slightly better than the laboratory. The accuracy and precision of the TcB measurement in this study was observed to be comparable to the standard of care laboratory test. PMID- 11389242 TI - Bacteremia, central catheters, and neonates: when to pull the line. AB - OBJECTIVES: Physicians who treat neonates who become bacteremic while dependent on central venous catheters face a serious and common dilemma. We sought 1) to evaluate the relationship between central venous catheter removal and outcome in bacteremic neonates, 2) to determine species of bacteria that are associated with an increased risk of infectious complications if the central catheter is not removed promptly, and 3) to provide evidence-based recommendations for central catheter management. METHOD: A retrospective cohort study of all neonates who had central venous access and developed bacteremia between July 1, 1995, and July 31, 1999, was conducted in the Duke University neonatal intensive care unit. RESULTS: The outcome for patients in whom the central catheter was not removed within 24 hours of organism identification was significantly worse (odds ratio = 9.8) than it was for those whose catheters were removed promptly. For patients who were infected with Staphylococcus aureus or with nonenteric Gram-negative rods, delayed removal of the central catheter was associated with complicated bacteremia. Catheter sterilization was attempted in 27 neonates who were infected with enteric Gram-negative rods; only 10 of these infants retained their catheters without infection-related complications. Infants who had 4 consecutive blood cultures that were positive for coagulase-negative staphylococcus (CoNS) were at significantly increased risk for end-organ damage and death, compared with infants who had 3 or fewer positive blood culture for CoNS (odds ratio = 29.58). CONCLUSIONS: Bacteremic infants experienced fewer infection-related complications when the central catheter was removed promptly. One positive blood culture for S aureus or a Gram-negative rod warrants central line removal in a neonate. Clinicians who are faced with a neonate who has 1 positive culture for CoNS may attempt medical management without central catheter removal, but documentation of subsequent negative blood cultures is crucial. Once a neonate has 3 positive blood cultures for CoNS, the central catheter should be removed.central line, neonate, bacteremia, bacteria, umbilical catheter, Broviac, percutaneous. PMID- 11389243 TI - Impact of prenatal diagnosis on survival and early neurologic morbidity in neonates with the hypoplastic left heart syndrome. AB - BACKGROUND: Prenatal echocardiography can identify the fetus that has complex congenital heart disease and may improve early management and surgical outcome. Prenatal diagnosis may be particularly beneficial to patients who have hypoplastic left heart syndrome (HLHS) and who are at risk for hypoxic-ischemic insult at presentation. OBJECTIVES: We sought to determine whether prenatal diagnosis reduces neurologic morbidity and operative mortality in patients who undergo palliative surgery for the HLHS. METHODS: Data from all patients who had HLHS, except for those with lethal genetic anomalies, and who were admitted to our institution between July 1992 and September 1997 were analyzed to assess the impact of prenatal diagnosis on preoperative management, neurologic morbidity, and surgical mortality. The primary outcome measures were hospital mortality and the incidence of adverse neurologic events (seizure or coma). RESULTS: There were 216 patients who had HLHS and were referred for surgical palliation, 79 (36.6%) of whom had been diagnosed prenatally. All patients who had been diagnosed prenatally were delivered in an advanced nursery and were started on prostaglandin E(1) on the first day of life. Patients whose HLHS was diagnosed postnatally were begun on prostaglandin E(1) later in life (median = day 2 [range = 1-28 days]). There were 4 preoperative deaths and 53 operative or postoperative deaths. Overall hospital mortality was 26.4% and did not differ between patients whose HLHS had been diagnosed prenatally and those whose HLHS had been diagnosed postnatally. With the use of multivariable analysis, prenatal diagnosis was associated with fewer adverse perioperative neurologic events in the patients whose HLHS had been diagnosed prenatally than in those whose HLHS had been diagnosed postnatally (odds ratio = 0.46). CONCLUSIONS: These data suggest that prenatal diagnosis has a favorable impact on treatment of patients who have HLHS and are undergoing staged palliation and reduces early neurologic morbidity. Prenatal diagnosis was not associated with reduced hospital mortality. It is possible that prenatal diagnosis may improve long-term neurologic outcome. PMID- 11389244 TI - Physicians' reliance on specialists, therapists, and vendors when prescribing therapies and durable medical equipment for children with special health care needs. AB - OBJECTIVES: Physicians who care for children with special (health care) needs (CWSN) often must prescribe therapies and/or specialized, durable medical equipment (DME). Given this responsibility and the increasing scrutiny of prescribing practices by various oversight agencies, understanding the extent to which pediatricians rely on their own expertise when prescribing therapies and DME is an important area of research. METHODS: As part of an ongoing investigation of physician preparedness for and practice in prescribing therapies, DME, or procedures for CWSN, we mailed surveys to practicing pediatricians in each of 2 states-Ohio and Mississippi-and to a senior resident at all identified pediatric residency-training programs. The surveys polled recipients as to who they would rely on themselves-specialists, therapists or vendors-to make prescription decisions for a variety of therapies and DME of increasing complexity. We report results as proportions of returned and completed questionnaires. Comparisons among the 3 groups (pediatricians from Ohio and Mississippi and residents) were made with the use of chi(2) analysis. RESULTS: For some categories of therapy and DME, physicians and residents reported that they would take an active role in prescription decisions, and their reliance on specialty consultation increased appropriately with the increasing complexity of the device or therapy. However, respondents generally seemed to share responsibility rather than rely on themselves as sole decision makers for most categories; fewer than one fourth took sole responsibility. Reliance on nonphysician health care providers was evident for all categories; in some cases, up to half of the respondents would allow therapists to take over these decisions, and a small but significant percentage of physicians would entrust DME prescription decisions to vendors alone. CONCLUSIONS: Our findings indicate that many practicing pediatricians and those in training may be unwilling to assume sole responsibility in prescribing and managing therapies and DME for CWSN. Although the number who would rely on consultation with specialists is somewhat reassuring, we found that a significant percentage would turn to nonphysician health care providers and even vendors to make these decisions in some cases, raising liability implications, conflict-of-interest issues, and quality-of-care issues. To protect themselves and their patients from fraud and inappropriate prescriptions and medical management, pediatricians must become increasingly conscientious about complying with American Medical Association guidelines and federal and state laws regarding initiation and supervision of therapies and DME. We offer some recommendations that may help to address this problem. PMID- 11389245 TI - Randomized trial comparing the efficacy of a novel manual breast pump with a standard electric breast pump in mothers who delivered preterm infants. AB - OBJECTIVE: The benefits of human milk for preterm infants are widely recognized, yet technological advances in milk expression have been slow. We compared the efficacy of a standard electric pump (EP; Egnell) used in 94% of United Kingdom neonatal units with a novel manual pump (MP; Avent ISIS) designed to operate more physiologically by simulating the infant's compressive action on the areola during breastfeeding. METHODS: We randomized 145 women who delivered infants of <35 weeks' gestation to use the MP or the EP and measured total milk volume expressed while using the randomized pump during the infant's hospital stay, pattern of milk output and creamatocrit of milk expressed during a test period in the second week, and pump characteristics by maternal questionnaire. RESULTS: Mothers who used the EP, who frequently double pumped, showed shorter expression times but produced no more milk than mothers who used the MP. When both pumped sequentially, however, mothers who used the MP showed significantly greater milk flow and total volume over 20 minutes. Creamatocrit was unaffected by pump type. The MP was rated significantly higher than the EP on 5 major characteristics. CONCLUSIONS: When compared on equal terms (sequential pumping), mothers who used the MP showed greater milk flow, perhaps reflecting more physiologic pump design. Even with double pumping, mothers who used the EP did not advantage their infants with greater milk production. We believe that this novel, effective MP, preferred by mothers and costing a fraction of the EP price, reflects a significant advance in milk expression for high-risk infants.breast pumps, randomized trial, preterm infants. PMID- 11389246 TI - Effect of pediatric surgical practice on the treatment of children with appendicitis. AB - OBJECTIVE: Acute appendicitis in children is managed by both general surgeons (GSs) and pediatric surgeons (PSs). Our objective was to investigate the economics of surgical care provided by either GSs or PSs for appendicitis. METHODS: The outcome of children within our state who underwent operative treatment for appendicitis (January 1994 to June 1997) by board-certified GSs were compared with the results of PSs. Data were sorted according to patient age and diagnosis according to the International Classification of Diseases, Ninth Revision. Analysis of variance was performed on continuous data, and chi(2) analysis was performed on nominal data; data are depicted as mean +/- standard error of the mean. RESULTS: GSs (n = 2178) managed older children when compared with PSs (n = 1018; 11.0 +/- 0.1 vs 9.1 +/- 0.1 years) and less frequently treated perforated appendicitis (18.8% vs 31.9%). Independent of diagnosis (simple or perforated appendicitis), younger children (0-4 years, 5-8 years, and 9-12 years) who were treated by PSs had a significantly shorter hospital stay and/or decreased hospital charge when compared with those who were treated by GSs. However, older children (13-15 years) seemed to have comparable outcomes. CONCLUSIONS: Younger children with appendicitis have reduced hospital days and charges when they are treated by PSs. PMID- 11389247 TI - A validity test of movie, television, and video-game ratings. AB - CONTEXT: Numerous studies have documented the potential effects on young audiences of violent content in media products, including movies, television programs, and computer and video games. Similar studies have evaluated the effects associated with sexual content and messages. Cumulatively, these effects represent a significant public health risk for increased aggressive and violent behavior, spread of sexually transmitted diseases, and pediatric pregnancy. In partial response to these risks and to public and legislative pressure, the movie, television, and gaming industries have implemented ratings systems intended to provide information about the content and appropriate audiences for different films, shows, and games. OBJECTIVE: To test the validity of the current movie-, television-, and video game-rating systems. DESIGN: Panel study. MEASURE: Participants used the KidScore media evaluation tool, which evaluates films, television shows, and video games on 10 aspects, including the appropriateness of the media product for children based on age. RESULTS: When an entertainment industry rates a product as inappropriate for children, parent raters agree that it is inappropriate for children. However, parent raters disagree with industry usage of many of the ratings designating material suitable for children of different ages. Products rated as appropriate for adolescents are of the greatest concern. The level of disagreement varies from industry to industry and even from rating to rating. Analysis indicates that the amount of violent content and portrayals of violence are the primary markers for disagreement between parent raters and industry ratings. CONCLUSIONS: As 1 part of a solution to the complex public health problems posed by violent and sexually explicit media products, ratings can have value if used with caution. Parents and caregivers relying on the ratings systems to guide their children's use of media products should continue to monitor content independently. Industry ratings systems should be revised with input from the medical and scientific communities to improve their reliability and validity. A single ratings system, applied universally across industries, would greatly simplify the efforts of parents and caregivers to use the system as well as the efforts of outside parties to monitor the use and validity of the system. PMID- 11389248 TI - Outcome in children receiving continuous venovenous hemofiltration. AB - OBJECTIVE: Continuous venovenous hemofiltration (CVVH) alone or with dialysis (D) has become an important supportive therapy for critically ill children with acute renal failure. Previous reports of pediatric patient outcome either mix CVVH/D with other renal replacement modalities or do not examine severity of illness. The current study examines only outcomes of children receiving CVVH/D using Pediatric Risk of Mortality (PRISM) scores to control for severity of illness. PATIENTS: Twenty-one patients (mean age: 8.8 +/- 6.3 years; mean weight: 28.3 +/- 20.8 kg) received 22 courses of CVVH/D. OUTCOMES: Nine (42.8%) of 21 patients survived. Nine (75%) of 12 deaths occurred within 25 days of pediatric intensive care unit (PICU) admission. Mean PRISM score at PICU admission and CVVH initiation were 13.1 +/- 5.8 and 15.4 +/- 8.9, respectively. Mean patient weight, age, PRISM score at PICU admission and at CVVH/D initiation, maximum pressor number, estimated glomerular filtration rate at CVVH/D initiation and change in mean airway pressure did not differ between survivors and nonsurvivors. The degree of fluid overload at CVVH/D initiation was significantly lower in survivors (16.4% +/- 13.8%) compared with nonsurvivors (34.0% +/- 21.0%), even when controlled for severity of illness by PRISM score. Mean cost of providing CVVH/D accounted for only 1% of total PICU cost per patient. CONCLUSIONS: The pattern of early multiorgan system failure and death, minimal relative cost of CVVH/D provision, and potential for improved outcome with initiation of CVVH/D at lesser degrees of fluid overload are factors that may support early initiation of CVVH/D in critically ill children with acute renal failure. PMID- 11389249 TI - Cisapride associated with QTc prolongation in very low birth weight preterm infants. AB - OBJECTIVE: No systematic study has been performed to evaluate the effect of cisapride on the QT interval in premature infants. Cisapride, which has recently been withdrawn by the Food and Drug Administration and is no longer an approved therapy, was commonly used for preterm infant care to improve the advance of enteral feedings and to reduce reflux and associated apnea. Our aim was to evaluate the effect of recommended doses of cisapride on the QT interval in this population. STUDY DESIGN: Prospective blinded evaluation of electrocardiogram for QT, JT, QTc, and JTc measurements in 25 preterm infants before and after cisapride administration. RESULTS: Twelve of 25 infants (48%) developed repolarization abnormalities with cisapride administration: 32% of the infants (8/25) studied had QTc prolongation (>/=0.450 seconds), whereas 10/25 had JTc prolongation (>/=0.360 seconds). Preterm infants <32 weeks significantly prolonged their QTc interval from 0.41 +/- 0.02 to 0.44 +/- 0.02. The QTc and/or JTc was prolonged in 54% of infants receiving 0.1 mg/kg/dose and 42% receiving 0.2 mg/kg/dose. CONCLUSIONS: The QTc and JTc interval significantly prolonged in preterm infants <32 weeks on the recommended dose of cisapride therapy. A QTc >/=0.450 seconds developed in 32% of infants treated with cisapride, whereas the JTc prolonged in 40%. A significant percentage of infants (54%) developed prolonged QTc intervals at a dose of 0.1 mg/kg/dose. From these data we conclude that there is a higher risk of prolongation of the QTc interval and risk of arrhythmias with greater prematurity. PMID- 11389250 TI - Infant sleep problems and postnatal depression: a community-based study. AB - OBJECTIVES: To describe infant sleep patterns and investigate relationships between infant sleep problems and maternal well-being in the community setting. DESIGN: Cross-sectional community survey. Setting. Maternal and Child Health Centers in 3 middle-class local government areas in Melbourne, Australia. PARTICIPANTS: Mothers of infants 6 to 12 months of age. MAIN OUTCOME MEASURES: Maternal well-being (Edinburgh Postnatal Depression Scale) and infant sleep problems (standardized maternal questionnaire). RESULTS: The survey was completed by 738 mothers (94% response rate), of whom 46% reported their infant's sleep as a problem. In the univariate analyses, sleep patterns characterizing a sleep problem included the infant sleeping in the parent's bed, being nursed to sleep, taking longer to fall asleep, waking more often and for longer periods overnight, and taking shorter naps. The same sleep patterns were associated with high depression scores and tended to increase as depression scores increased. Because of positive skew, the Edinburgh Postnatal Depression Score was analyzed in 3 categories (<10, 10-12, and >12) using validated cutoff scores from community and clinical studies. Fifteen percent of mothers scored above 12 on the depression scale, indicating probable clinical depression, and 18% scored between 10 and 12, indicating possible clinical depression. After adjusting for potential confounders and factors significant in the univariate analyses, maternal report of an infant sleep problem remained a significant predictor of a depression score >12 (odds ratio: 2.13; 95% confidence interval: 1.27,3.56) and >10 (odds ratio: 2.88; 95% confidence interval: 1.93,4.31). However, mothers reporting good sleep quality, despite an infant sleep problem, were not more likely to suffer depression. CONCLUSIONS: Maternal report of infant sleep problems and depression symptoms are common in middle-class Australian communities. There is a strong association between the 2, even when known depression risk factors are taken into account. Maternal report of good sleep quality attenuates this relationship. Appropriate anticipatory guidance addressing infant sleep could potentially decrease maternal report of depressive symptoms. PMID- 11389251 TI - Feasibility of tidal volume-guided ventilation in newborn infants: a randomized, crossover trial using the volume guarantee modality. AB - BACKGROUND AND AIM: Volume guarantee (VG) is a new composite mode of pressure limited ventilation, available on the Drager Babylog 8000 ventilator, which allows the clinician to set a target mean tidal volume to be delivered while still maintaining control over peak airway pressures. This study aimed to investigate the feasibility and efficacy of this mode of ventilation in premature newborn infants with respiratory distress syndrome (RDS). METHODS: Two groups of infants were studied: those receiving synchronized intermittent positive pressure ventilation (SIPPV) in early phase of RDS (group 1) and those in recovery phase of RDS being weaned from artificial ventilation through synchronized intermittent mandatory ventilation (SIMV; group 2). Both groups of infants were studied over a 4-hour period. Before the start of the study, the infants were either receiving SIPPV (group 1) or SIMV (group 2). Infants in group 1 were randomized to either continue on SIPPV for the first hour of the study or to receive SIPPV plus VG for the first hour. Subsequently, the 2 modes were used alternately for the remaining three 1-hour periods. Similarly, infants in group 2 were randomized to either continue on SIMV for the first hour of the study or to receive SIMV plus VG for the first hour. Data on ventilation parameters and transcutaneous carbon dioxide and oxygen were collected continuously. RESULTS: Forty infants were studied, 20 in each group. The mean (standard error) gestational age was 27.9 (0.3) weeks; birth weight was 1064 (60) g. No adverse events were observed during the study. Fractional inspired oxygen during SIMV plus VG was 0.31 (0.3); during SIMV, 0.31 (0.3); during SIPPV plus VG, 0.41 (0.4); and during SIPPV, 0.40 (0.4). Transcutaneous carbon dioxide pressure during SIMV plus VG was 6.0 (2.2) kPa; during SIMV, 5.9 (2.2) kPa; during SIPPV plus VG, 6.4 (2.9) kPa; and during SIPPV, 6.4 (2.8) kPa. Transcutaneous partial pressure of oxygen during SIMV plus VG was 8.4 (8.7) kPa; during SIMV, 8.6 (8.8) kPa; during SIPPV plus VG, 7.6 (4.0) kPa; and during SIPPV, 7.7 (4.2) kPa. None of these differences was statistically significant. The mean (standard error) peak inspiratory pressure used during SIMV was 17.1 (3.4) cm of water; during SIMV plus VG, 15.0 (7.5) cm of water; during SIPPV plus VG, 17.1 (9.3) cm of water; and during SIPPV, 18.7 (8.3) cm of water. The mean airway pressure during SIMV plus VG was 6.5 (3.1) cm of water; during SIMV, 6.9 (2.8) cm of water; during SIPPV plus VG, 9.6 (4.5) cm of water; and during SIPPV, 9.8 (4.6) cm of water. CONCLUSION: VG seems to be a stable and feasible ventilation mode for neonatal patients and can achieve equivalent gas exchange using statistically significant lower peak airway pressures both during early and recovery stages of RDS.ventilation, airway pressure, volume guarantee, tidal volume. PMID- 11389252 TI - The role of protein C, protein S, and resistance to activated protein C in Legg Perthes disease. AB - OBJECTIVES: It has been hypothesized that Legg-Perthes disease is caused by repeated vascular interruptions of the blood supply to the proximal femur, which are precipitated by coagulation system abnormalities. To test this theory, we conducted a case-control study among 57 patients with Legg-Perthes disease and an equal number of community controls. We measured protein C and protein S and resistance to activated protein C (APC-R) from plasma. STUDY DESIGN: Participants were placed into 1 of 3 mutually exclusive categories based on the control distribution: 1) normal, defined as either above or within 1 standard deviation below the expected mean; 2) low normal, defined as between 1 and 2 standard deviations below the expected mean; and 3) low, defined as >2 standard deviations below the expected mean. DNA was analyzed to determine the presence of a point mutation in the factor V gene that causes APC-R. RESULTS: We observed a statistically significant increased risk of Legg-Perthes disease with decreasing levels of protein C and a nearly significant increased risk with decreasing levels of protein S. The factor V gene defect was present in 5 (9%) of 55 cases and 3 (5%) of 56 controls (odds ratio 1.8, 95% confidence interval: 0.4-7.7), but the mean level on the APC-R plasma test was similar for cases and controls. Nine cases and 1 control had 2 low normal or low test results (odds ratio 13.0, 95% confidence interval: 2.2-75). CONCLUSIONS: Our results support the belief that abnormalities of the coagulation system leading to a thrombophilic state play a role in Legg-Perthes disease; however, larger studies are needed before definitive recommendations for coagulation testing can be made. PMID- 11389253 TI - Effects of medicaid managed care on quality: childhood immunizations. AB - BACKGROUND: Underimmunization is distributed unevenly across populations, concentrated among the impoverished. Managed care has stimulated the development of quality indicators such as immunization rates to assess health status of populations. OBJECTIVE: To determine if enrollment in Medicaid managed care (MMC) improves quality of health care as reflected by immunization rates when compared with fee-for-service Medicaid (FFSM). DESIGN. Prospective cohort study of infants born between May 1994 and April 1995 with a 24-month follow-up period. SETTING: Urban teaching hospital and surrounding ambulatory settings. PARTICIPANTS: Consecutive sample of infants (n = 644) enrolled in MMC or FFSM. Ninety-two percent of eligible patients were enrolled, and 87% completed follow-up. MAIN OUTCOME MEASURE: Up-to-date immunization status. RESULTS: Seventy-three percent of the MMC and 72.4% of the FFSM patients were up-to-date on their immunizations: relative risk 1.01, (95% confidence interval [CI] 0.87, 1.17). No differences were found in age at immunization between the MMC and FFSM groups. After adjusting for other factors in multivariate analysis, insurance status remained unassociated with immunization status: adjusted odds ratio (OR) 1.04, (95% CI: 0.90, 1.10). Factors associated with up-to-date immunization included firstborn child, OR 2.28 (95% CI: 1.45, 3.60) and adequate maternal prenatal care, OR 2.24 (95% CI: 1.44, 3.48). Variables characterizing children less likely to be adequately immunized included father living in home with child, OR 0.53 (95% CI: 0.33, 0.85) and using private office-based primary care, OR 0.39 (95% CI: 0.23, 0.63). CONCLUSIONS: Enrollment in MMC did not improve rates of immunizations when compared with FFSM. PMID- 11389254 TI - Successful management of tracheotomized patients with chronic saliva aspiration by use of constant positive airway pressure. AB - OBJECTIVE: Management of chronic aspiration of saliva is a challenge to clinicians. The purpose of this report is to review the clinical course of 3 patients with tracheotomy who we have followed for at least 1 year and who have received constant positive airway pressure (CPAP) as a primary treatment for ongoing aspiration of saliva. METHODS: Retrospective chart review. RESULTS: We present here 3 patients with chronic congestion and persistent hypoxemia in whom a diagnosis of chronic aspiration of saliva was established by use of radionuclide salivagram. Each of these children had tracheotomy for treatment of airway obstruction. In an attempt to decrease chronic aspiration of saliva, we instituted constant positive pressure via tracheotomy. Repeat radionuclide salivagram performed on CPAP demonstrated a marked decrease in saliva aspiration. All patients experienced improvement in clinical symptoms and required only rare subsequent hospitalizations for respiratory disease. CONCLUSION: We suggest, based on this case series, that CPAP administered via a tracheotomy is an acceptable means of managing chronic salivary aspiration and that it may decrease respiratory complications in such patients. PMID- 11389255 TI - Renal function after pediatric cardiac transplantation: the effect of early cyclosporin dosage. AB - BACKGROUND: There is little data on renal function in pediatric heart transplant recipients. Early rejection is a major concern and most units run high cyclosporin A (CyA) levels during the 2 to 3 months after transplantation. We sought to document long-term renal function after transplantation and to assess influence of early CyA levels. METHODS: We reviewed all of our pediatric transplants between June 1985 and August 1998 who survived longer than 6 months (n = 54). Glomerular filtration rate (GFR) was estimated at 1, 2, 4, and 8 years posttransplantation using the Schwartz formula: GFR (mL/min/1.73m(2)) = [Ht(cm)/creatinine(micromol/L)] x X We also analyzed whether change in renal function correlated with trough CyA levels. RESULTS: Median age at transplant was 4 years and median follow-up was 5 years. Survival rates were 87% at 1 year and 80% at 5 years. Mean GFR pretransplant was 79 +/- 19 mL/min/1.73 m(2), reflecting prerenal impairment. One year later, mean GFR was 72 mL/min/1.73 m(2); after 2 years it was 65 mL/min/1.73 m(2), after 4 years (n = 35) it was 60 mL/min/1.73 m(2), and after 8 years (n = 14) it was 57 mL/min/1.73 m(2). CyA levels during the first 2 months correlated with the change in GFR during the first year (r(2) = 0.21). CONCLUSIONS: This study demonstrates for the first time that decline in renal function after heart transplantation correlates with early CyA exposure; this dysfunction persists even when CyA doses are subsequently reduced. PMID- 11389256 TI - Child health status, neurodevelopmental outcome, and parental satisfaction in a randomized, controlled trial of nitric oxide for persistent pulmonary hypertension of the newborn. AB - OBJECTIVE: To describe health and neurodevelopmental outcomes and parental satisfaction with hospital care among surviving intervention and control enrollees in a randomized, controlled trial of nitric oxide for persistent pulmonary hypertension of the newborn (PPHN). METHODS: All surviving enrollees 1 to 4 years of age were eligible for follow-up. Outcomes were assessed by telephone using a trained interviewer and standardized instruments. Domains assessed included parental report of specific conditions and hospital use, rating of general health, cognitive and motor development, behavior problems, temperament, and satisfaction with the hospital stay. Fisher's exact test and the Wilcoxon rank sum test assessed differences between intervention and control infants. RESULTS: Interviews were completed on 60 of 83 survivors (72%). Eighteen families (22%) could not be located, 2 (2%) were non-English-speaking, and 3 (4%) declined participation. No postdischarge deaths were ascertained. Among those interviewed, race, income, and education of parents of intervention and controls were comparable, as were entry oxygenation index, extracorporeal oxygenation utilization, and days of hospitalization. No differences were found in pulmonary, neurologic, cognitive, behavioral, or neurosensory outcomes; hospital readmission rates; or parental ratings of child's health. The overall neurologic handicap rate was 15%. The rate of hearing deficit was 7%. The rate of significant behavioral problems was 26%. Levels of satisfaction expressed were high for each group. No differences in parental ratings were found between the 2 groups. CONCLUSIONS: No adverse health or neurodevelopmental outcomes have been observed among infants treated with nitric oxide for PPHN. The parents of the critically ill infants enrolled in our clinical trial welcomed their child's inclusion and all expressed satisfaction with the care that their child received while at a tertiary care hospital. Enrollment in either arm of this randomized, controlled trial did not seem to affect parental satisfaction with the hospital care that their child received. PMID- 11389257 TI - Short-term outcomes after acute treatment of pediatric asthma. AB - CONTEXT: The short-term course of pediatric patients after emergency department (ED) treatment for acute asthma has not been comprehensively documented; most previous studies have limited outcomes to ED length-of-stay, hospital admission, and relapse. OBJECTIVE: To describe symptom persistence, medication use, functional disability, follow-up, and relapse in these children in the 2 weeks after acute treatment and ED discharge. DESIGN: Randomly selected, prospective cohort from September 1996 to August 1997; follow-up telephone interviews at 1 and 2 weeks. Setting. A large, inner-city children's hospital emergency department. Patients. Random sample of pediatric asthma visits requiring ED treatment but not admission; 457 were eligible, 388 with complete follow-up (85%); final sample included 367 patients after multiple visits deleted. MAIN OUTCOME MEASURES: Details of symptom persistence, functional disability, medication use, relapse, and routine follow-up. RESULTS: Results included significant morbidity: 23% (95% confidence interval [CI]: 19, 27) with cough and 12% (95% CI: 9, 15) with wheeze persistent at 2 weeks; 20% (95% CI: 16, 24) with decreased activity at 1 week; 45% (95% CI: 39, 51) missed >2 and 24% (95% CI: 19, 29) >/=5 days of school or day care; 17% (95% CI: 13, 21) spent >/=3 days in bed; 54% (95% CI: 47, 60) of caretakers missed at least 1 and 18% (95% CI: 13, 24) missed >2 days of school or work; and 32% (95% CI: 28, 38) of patients were still using greater than baseline medication at 2 weeks. Reported relapse rates were averaged at 13% (95% CI: 10, 17) with 3% (95% CI: 1, 5) admitted. Routine office follow-up was poor: 29% (95% CI: 25, 34) had had a visit; 48% (95% CI: 43, 54) reported no visit/none planned. CONCLUSIONS: A considerable proportion of inner city pediatric patients discharged from the hospital from the ED after standard treatment for acute asthma had poor short-term outcomes. Conventional markers of successful ED treatment, such as avoiding hospital admission or relapse, do not adequately describe outcomes of acute care. The patient-oriented measures described here may provide more useful indicators of outcome in the evaluation of acute asthma care. PMID- 11389259 TI - Depiction of alcohol, tobacco, and other substances in G-rated animated feature films. AB - OBJECTIVE: To quantify and characterize the depiction of alcohol, tobacco, and other substances in G-rated animated feature films. METHOD: The content of all G rated animated feature films released in theaters between 1937 and 2000, recorded in English, and available on videocassette in the United States by October 31, 2000, was reviewed for portrayals of alcohol, tobacco, and other substances and their use. Duration of scenes depicting alcohol, tobacco, or other substances; type of characters using them (good, neutral, or bad); and correlation of amount and type used with character type and movie type were evaluated. RESULTS: Of the 81 films reviewed, 38 films (47%) showed alcohol use (mean exposure: 42 seconds per film; range: 2 seconds to 2.9 minutes) and 35 films (43%) showed tobacco use (mean exposure: 2.1 minutes per film; range: 2 seconds to 10.5 minutes). Analysis of time trends showed a significant decrease in both tobacco and alcohol use over time (both corrected for total screen duration and uncorrected.) No films showed the use of illicit drugs, although 3 films showed characters consuming a substance that transfigured them and 2 films showed characters injected with a drug. Analysis of the correlation of alcohol and tobacco depiction revealed several scenes in which alcohol and tobacco were shown in use in the same scene and that bar scenes in these movies depict a significant amount of drinking, smoking, and violence. Three films contained a message that a character should stop smoking but none contained messages about restricting consumption of alcohol. CONCLUSIONS: The depiction of alcohol and tobacco use in G-rated animated films seems to be decreasing over time. Nonetheless, parents should be aware that nearly half of the G-rated animated feature films available on videocassette show alcohol and tobacco use as normative behavior and do not convey the long-term consequences of this use. PMID- 11389258 TI - Parental monitoring: association with adolescents' risk behaviors. AB - CONTEXT: Contemporary threats to adolescents' health are primarily the consequence of risk behaviors and their related adverse outcomes. Identifying factors associated with adolescents' risk behaviors is critical for developing effective prevention strategies. A number of risk factors have been identified, including familial environment; however, few studies have examined the impact of parental monitoring. OBJECTIVE: To examine the influence of less perceived parental monitoring on a spectrum of adolescent health-compromising behaviors and outcomes. Design. Survey. SETTING: A family medicine clinic. Participants. To assess eligibility, recruiters screened a sample of 1130 teens residing in low income neighborhoods. Adolescents were eligible if they were black females, between the ages of 14 and 18 years, sexually active in the previous 6 months, and provided written informed consent. Most teens (n = 609) were eligible, with 522 (85.7%) agreeing to participate. MAIN OUTCOME MEASURES: Variables in 6 domains were assessed, including: sexually transmitted diseases, sexual behaviors, marijuana use, alcohol use, antisocial behavior, and violence. RESULTS: In logistic regression analyses, controlling for observed covariates, adolescents perceiving less parental monitoring were more likely to test positive for a sexually transmitted disease (odds ratio [OR]: 1.7), report not using a condom at last sexual intercourse (OR: 1.7), have multiple sexual partners in the past 6 months (OR: 2.0), have risky sex partners (OR: 1.5), have a new sex partner in the past 30 days (OR: 3.0), and not use any contraception during the last sexual intercourse episode (OR: 1.9). Furthermore, adolescents perceiving less parental monitoring were more likely to have a history of marijuana use and use marijuana more often in the past 30 days (OR: 2.3 and OR: 2.5, respectively); a history of alcohol use and greater alcohol consumption in the past 30 days (OR: 1.4 and OR: 1.9, respectively); have a history of arrest (OR: 2.1); and there was a trend toward having engaged in fights in the past 6 months (OR: 1.4). CONCLUSIONS: The findings demonstrate a consistent pattern of health risk behaviors and adverse biological outcomes associated with less perceived parental monitoring. Additional research needs to focus on developing theoretical models that help explain the influence of familial environment on adolescent health and develop and evaluate interventions to promote the health of adolescents. PMID- 11389260 TI - Sleep problems seldom addressed at two general pediatric clinics. AB - OBJECTIVE: Sleep disorders can cause substantial morbidity but often remain undiagnosed among adults. We identified a series of children with sleep-related symptoms and reviewed medical chart notes for the previous 2 years to determine how often sleep problems had been addressed. DESIGN: Observational. Setting. Two university-affiliated but community-based general pediatrics clinics. PATIENTS: Children, ages 2.0 to 13.9 years, with clinic appointments. MEASURES: Parental and child responses to a validated Pediatric Sleep Questionnaire (PSQ) were used to identify patients at risk for chronic sleep-disordered breathing, periodic leg movements during sleep, insomnia, or excessive daytime sleepiness. Chart notes written within the previous 2 years were searched for sleep-related symptoms, diagnoses, or treatments. RESULTS: A total of 830 questionnaires were completed; 1395 chart notes of 86 symptomatic participants (mean age: 6.6 +/- 3.1 years; 51% male) with 103 identified sleep problems were reviewed. Fewer than 15% of patients had current chart notes that mentioned any of the PSQ-defined sleep problems; diagnoses were mentioned for 2 of 86 patients and no treatments were discussed. Among the 103 sleep problems, only 16 received mention in any past or current note; 10 had led to a diagnosis; 4 had led to intervention; and 3 were treated in a manner likely to be effective. Seventy-four of the sleep problems (72%) occurred in children whose charts did mention something about sleep, but such notations rarely related to concerns uncovered by the PSQ. CONCLUSIONS: Children with PSQ-identified sleep problems at 2 general pediatrics clinics seldom had these problems addressed, diagnosed, or treated, despite discussions about some aspect of their sleep in the large majority of cases. These findings support expansion of clinician and parent education about sleep disorders in children. PMID- 11389261 TI - Iron deficiency and cognitive achievement among school-aged children and adolescents in the United States. AB - CONTEXT: Iron deficiency anemia in infants can cause developmental problems. However, the relationship between iron status and cognitive achievement in older children is less clear. OBJECTIVE: To investigate the relationship between iron deficiency and cognitive test scores among a nationally representative sample of school-aged children and adolescents. DESIGN: The National Health and Nutrition Examination Survey III 1988-1994 provides cross-sectional data for children 6 to 16 years old and contains measures of iron status including transferrin saturation, free erythrocyte protoporphyrin, and serum ferritin. Children were considered iron-deficient if any 2 of these values were abnormal for age and gender, and standard hemoglobin values were used to detect anemia. Scores from standardized tests were compared for children with normal iron status, iron deficiency without anemia, and iron deficiency with anemia. Logistic regression was used to estimate the association of iron status and below average test scores, controlling for confounding factors. RESULTS: Among the 5398 children in the sample, 3% were iron-deficient. The prevalence of iron deficiency was highest among adolescent girls (8.7%). Average math scores were lower for children with iron deficiency with and without anemia, compared with children with normal iron status (86.4 and 87.4 vs 93.7). By logistic regression, children with iron deficiency had greater than twice the risk of scoring below average in math than did children with normal iron status (odds ratio: 2.3; 95% confidence interval: 1.1-4.4). This elevated risk was present even for iron-deficient children without anemia (odds ratio: 2.4; 95% confidence interval: 1.1-5.2). CONCLUSIONS: We demonstrated lower standardized math scores among iron-deficient school-aged children and adolescents, including those with iron deficiency without anemia. Screening for iron deficiency without anemia may be warranted for children at risk. PMID- 11389262 TI - Physical activity and bone measures in young children: the Iowa bone development study. AB - OBJECTIVES: Physical activity has a beneficial effect on bone development in circumpubertal children, although its effect on younger children is uncertain. In this cross-sectional study, we examined associations between physical activity and bone measures in 368 preschool children (mean age: 5.2 years, range: 4-6 years). DESIGN: Physical activity was measured using 4-day accelerometry readings, parental report of children's usual physical activity, and parental report of children's hours of daily television viewing. Total body and site specific bone mineral content and area bone mineral density (BMD) were measured by dual energy radiograph absorptiometry. RESULTS: After adjustment for age and body size, accelerometry measures of physical activity and parental report of usual physical activity were consistently and positively associated with bone mineral content and BMD in both boys and girls (r = 0.15-0.28). Television viewing was inversely associated with hip BMD in girls (r = -0.15). The proportion of variance in bone measures explained by physical activity in linear regression models ranged from r(2) = 1.5% to 9.0%. In all of these models except total body BMD, at least 1 and often several of the physical activity variables entered as independent predictors. Activity variables most likely to enter the regression models were vigorous physical activity (as determined by accelerometry) and parental ranking of child's usual physical activity. CONCLUSIONS: Findings indicate that there are statistically significant and, perhaps important, associations between physical activity and bone measures during early childhood, well ahead of the onset of peak bone mass. This would suggest that intervention strategies to increase physical activity in young children could contribute to optimal bone development. PMID- 11389263 TI - Snoring during early childhood and academic performance at ages thirteen to fourteen years. AB - OBJECTIVES: Obstructive sleep apnea syndrome in young children is associated with an adverse effect on learning. However, the long-term impact of sleep-disordered breathing (SDB) during early childhood on learning remains unknown. METHODS: Questionnaires were mailed to seventh and eighth graders attending public schools whose class ranking was either in the top 25% (high performance [HP]) or bottom 25% of their class (low performance [LP]), and who were matched for age, gender, race, school, and street of residence. Snoring frequency and loudness at 2 to 6 years of age, tonsillectomy and adenoidectomy (T&A) for snoring or recurrent infection, school grades, and parental smoking and snoring were assessed. RESULTS: The questionnaire response rate was 82.8%. Because of ongoing ring, 13 responders were excluded, such that 1588 questionnaires could be analyzed (797 in LP and 791 in HP group). Frequent and loud snoring during early childhood was reported in 103 LP children (12.9%) compared with 40 HP children (5.1%; odds ratio: 2.79; confidence interval: 1.88-4.15). Furthermore, 24 LP and 7 HP children underwent T&A for snoring (odds ratio: 3.40; confidence interval: 1.47 7.84), while 21 LP and 19 HP children required surgery for recurrent tonsillitis. CONCLUSIONS: Children with lower academic performance in middle school are more likely to have snored during early childhood and to require T&A for snoring compared with better performing schoolmates. These findings support the concept that SDB-associated neurocognitive morbidity may be only partially reversible or that a "learning debt" may develop with SDB during early childhood and hamper subsequent school performance. PMID- 11389264 TI - Prothrombotic disorders and abnormal neurodevelopmental outcome in infants with neonatal cerebral infarction. AB - BACKGROUND AND PURPOSE: The aim of this study was to evaluate the occurrence of prothrombotic disorders in a well-characterized cohort of infants with neonatal stroke and to document any association of prothrombotic disorders with the type of infarct seen on magnetic resonance imaging (MRI) and clinical outcome. METHODS: Twenty-four infants with perinatal cerebral infarction confirmed by neonatal MRI were enrolled in the study. All the infants and, when possible, both parents were tested to identify inherited and acquired prothrombotic disorders. RESULTS: None of the infants had a significant bleeding diathesis, but 10 (42%) had at least 1 prothrombotic risk factor. Five children showed heterozygosity for factor V Leiden, and 6 had high factor VIIIc concentrations. There was a striking association between the occurrence of these abnormalities and both the presence of cerebral hemorrhage on MRI and poor neurologic outcome. Eight of the 11 patients (73%) with hemiplegia or global developmental delay had factor V Leiden and/or raised factor VIIIc, whereas only 1 of the 13 patients (8%) with normal outcome had any prothrombotic risk factors. In particular, all 5 infants with factor V Leiden had hemiplegia, compared with only 4 of the 19 infants without factor V Leiden (21%). CONCLUSIONS: These data suggest that the presence of prothrombotic risk factors and, in particular, of the factor V Leiden mutation, is significantly associated with poor outcome after perinatal cerebral infarction. PMID- 11389265 TI - Access to orthopedic care for children with medicaid versus private insurance in California. AB - OBJECTIVE: To compare the availability of timely orthopedic care to a child with a fractured arm insured by Medi-Cal (California state Medicaid) and by private insurance. STUDY DESIGN: Fifty randomly chosen offices of orthopedic surgeons were telephoned with the following scenario: "My 10-year-old son broke his arm last week during a vacation" followed by a request for an appointment that week. Each office was called twice with an identical script except for insurance status: once with Medi-Cal and once with private insurance. RESULTS: All 50 offices offered an appointment to see the child with private insurance within 7 days. Only 1 of the same 50 offices offered an appointment to see the child with Medi-Cal within 7 days. Of the offices that would not see a child with Medi-Cal, 87% were unable to recommend an orthopedic office that accepted Medi-Cal. CONCLUSIONS: Timely access to orthopedic care was available in 100% of offices polled to a child with private insurance versus in 2% of offices to a child with Medi-Cal. This is a significant difference. Lack of timely orthopedic care may result in poor outcome, ie, if a fracture is not properly aligned in the first few weeks, a permanent deformity may result. Although causation cannot be established from this study, we suspect that Medi-Cal reimbursement rates below the cost of office overhead may be of significance. Although federal guidelines require that payments must be sufficient to enlist enough providers so that services to Medi-Cal recipients are available to the same extent as those available to the general population, this study finds that that children with Medi-Cal insurance have significantly less access to timely orthopedic care. PMID- 11389266 TI - Lung transplantation and cystic fibrosis: the psychosocial toll. PMID- 11389267 TI - Unnatural selection. PMID- 11389268 TI - Privacy and its regulation: too much too soon, or too little too late. PMID- 11389269 TI - Postnatal steroids and neurodevelopmental outcomes: a problem in the making. PMID- 11389270 TI - "You are hereby commanded to appear": pediatrician subpoena and court appearance in child maltreatment. AB - OBJECTIVE: To determine the frequency of court appearance by pediatricians evaluating child abuse and neglect cases and to identify case characteristics associated with actual court appearance or case adjournment. DESIGN: Retrospective review of subpoenas received between 1995 and 1999 for child maltreatment cases personally evaluated by 2 pediatricians during the years 1995 to 1998. Information was collected regarding patient age, gender, race/ethnicity, type of suspected maltreatment, date of evaluation, date of subpoena, type of court hearing, whether the pediatrician actually testified in court, and legal outcomes. Case characteristics were compared between pediatricians and were used to predict physician appearance and case continuance or adjournment in logistic regression models. RESULTS: Four hundred forty-five subpoenas concerning 260 patients were received. Although significant differences were noted between the pediatricians in type of abuse, no differences were found in patient age, gender, ethnicity or legal outcomes. The pediatricians received subpoenas in <15% of child maltreatment cases, and <5% of children seen resulted in the physician being required to actually appear in court. No case characteristics significantly predicted court appearance or case continuance or adjournment. CONCLUSIONS: Although pediatricians are sometimes subpoenaed to appear in court to explain the medical evaluation and the needs of the child in cases of child abuse and neglect, most court cases were continued, adjourned, or settled before physician testimony. Most subpoenas did not result in the pediatrician going to court, and it is unclear which child factors may predict court involvement. Pediatricians can take steps to minimize (but not eliminate) the potential dissatisfaction and inconvenience associated with receiving and responding to subpoenas in child maltreatment cases. PMID- 11389271 TI - A randomized trial comparing povidone-iodine to a chlorhexidine gluconate impregnated dressing for prevention of central venous catheter infections in neonates. AB - Neonates who require a central venous catheter (CVC) for prolonged vascular access experience high rates of catheter-related bloodstream infection (CRBSI). PURPOSE: A multicenter randomized clinical trial was undertaken to ascertain the efficacy of a novel chlorhexidine-impregnated dressing (Biopatch Antimicrobial Dressing) on the CVC sites of neonates for the prevention of catheter tip colonization, CRBSI, and bloodstream infection (BSI) without a source. Setting. Six level III neonatal intensive care units. Patients Studied. Neonates admitted to study units who would require a CVC for at least 48 hours. METHODS: Eligible infants were randomized before catheter placement to 1 of the 2 catheter site antisepsis regimens: 1) 10% povidone-iodine (PI) skin scrub, or 2) a 70% alcohol scrub followed by placement of a chlorhexidine-impregnated disk over the catheter insertion site. A transparent polyurethane dressing (Bioclusive Transparent Dressing) was used to cover the insertion site in both study groups. Primary study outcomes evaluated were catheter tip colonization, CRBSI, and BSI without an identified source. RESULTS: Seven hundred five neonates were enrolled in the trial, 335 randomized to receive the chlorhexidine dressing and 370 to skin disinfection with PI (controls). Neonates randomized to the antimicrobial dressing group were less likely to have colonized CVC tips than control neonates (15.0% vs 24.0%, relative risk [RR]: 0.6 95% confidence interval [CI]: 0.5-0.9). Rates of CRBSI (3.8% vs 3.2%, RR: 1.2, CI: 0.5-2.7) and BSI without a source (15.2% vs 14.3%, RR: 1.1, CI: 0.8-1.5) did not differ between the 2 groups. Localized contact dermatitis from the antimicrobial dressing, requiring crossover into the PI treatment group, occurred in 15 (15.3%) of 98 exposed neonates weighing /=12 months of age at the time that data were collected were categorized as being fully immunized at 12 months if they had received the same vaccines before their first birthday. To assess the effect of type of poliovirus vaccines on these outcomes, study patients were classified as being in an IPV or OPV group based on the initial type of vaccine received. Logistic regression was used to calculate the odds ratios (ORs) and 95% confidence intervals (CIs) for IPV as a predictor of being fully immunized at 8 and 12 months of age, after adjusting for race/ethnicity of the patient, maternal education level, year of birth, and method of payment for vaccines. In addition, the effect of clustering of children within practices was accounted for by the use of generalized estimation equation techniques. RESULTS: Data were analyzed on 13 520 children from 177 practices in 42 states; 79.4% of patients were fully immunized at 8 months of age, and 88.7% of those eligible were fully immunized at 12 months of age. A total of 6910 patients (51.1%) were classified as OPV recipients, wheras 5282 (39.1%) received IPV. In addition, 1328 children (9.8%) were documented as having received poliovirus vaccine, but the particular type could not be determined. Compared with OPV recipients and after controlling for the confounding variables and the effect of clustering within practices, children in the IPV group were as likely as were OPV recipients to be fully immunized at 8 months of age (OR: 1.04; 95% CI: 0.88,1.23). At 12 months of age, the OR for IPV as a predictor of being fully immunized was 1.08 (95% CI: 0.90,1.30). When compared with OPV recipients, adjusted ORs for children in the undetermined poliovirus vaccine type group being fully immunized at 8 and 12 months of age were 0.84 (95% CI: 0.68,1.04) and 0.84 (95% CI: 0.67,1.07), respectively. CONCLUSIONS: The results of this national study indicate that the implementation of an IPV-containing poliovirus vaccine schedule has not had an adverse effect on the immunization status of young children who were vaccinated in the offices of practicing pediatricians. PMID- 11389289 TI - The effects of environmental tobacco smoke on health services utilization in the first eighteen months of life. AB - OBJECTIVE: To examine the effects of environmental tobacco smoke (ETS) on health services use in Chinese infants with nonsmoking mothers. DESIGN: Prospective, population-based birth cohort. SETTING: General population of Hong Kong in 1997 1998. PARTICIPANTS: A total of 8327 parent-infant pairs who were followed up for 18 months. MAIN OUTCOME MEASURES: Doctor consultations and hospitalizations. Results. After adjusting for the age, education level, and employment status of mothers-as well as infants' birth weight, method of delivery, breastfeeding status, and birth order-ETS exposure through the mother in utero was positively associated with higher consultation (adjusted odds ratio [OR]: 1.26; 95% confidence interval [CI]: 1.14, 1.39) and hospitalization (OR: 1.18; 95% CI: 1.05, 1.31) use in infants with nonsmoking mothers attributable to any illness. In addition, postnatal exposure to ETS at home was linked to higher rates of hospitalizations for any illness compared with nonexposed infants (OR: 1.12; 95% CI: 1.00, 1.25), although the relationship did not hold for outpatient consultation visits. The OR for higher hospital use in infants exposed to 2 or more smokers at home was 1.30 (95% CI: 1.08, 1.58). CONCLUSIONS: The use of tobacco products by household members, even among nonsmoking mothers, has an enormous adverse impact on the health of children, as well as increases health services use and cost. The present data support the revision of public policy to reflect an evidence-based approach to the promotion of smoking cessation in all household members during and after pregnancy. environmental tobacco smoke, health services, infants. PMID- 11389290 TI - Body image and dieting behavior in cystic fibrosis. AB - OBJECTIVE: To examine the relationship between pulmonary function, nutritional status, body image, and eating attitudes in children with cystic fibrosis (CF) compared with healthy controls. METHOD: Seventy-six children with CF (39 girls) and 153 healthy control children (82 girls) were recruited. All children were between 7 and 12 years of age. After being weighed and measured, participants undertook a structured 1-to-1 interview. Four measures were used to assess body image: body size (perception and satisfaction) were ascertained using the Children's Body Image Scale (CBIS), which uses photographs of children of various body mass index (BMI) representative of the range of BMI percentiles for children 7 to 12 years of age. Body size satisfaction was measured by the response to the questions, "Do you think your body is 1) much too thin, 2) too thin, 3) just right, 4) too fat, and 5) much too fat?" Body weight satisfaction was measured by the question, "Would you like your body to be 1) much thinner, 2) a little bit thinner, 3) stay the same, 4) a little bit fatter, and 5) much fatter?" Global self-esteem was measured using the children's version of the Rosenberg Self Esteem Scale and Body Esteem Scale using a 24-item scale. Dieting behavior was measured by asking directly about previous weight control behaviors, use of the Dutch Eating Behavior-Restraint Scale (DEBQ-R), and, in children who acknowledged previous dieting behavior, the Children's Eating Attitude Test (ChEAT) was additionally administered. Results. Both girls and boys with CF had significantly reduced BMI percentiles compared with control children. Boys with CF did not have a significantly different BMI compared with girls with CF. There were significant positive correlations between forced expiratory volume in 1 second (FEV(1)) (% of predicted) and BMI percentile in girls (r =.35) and boys (r =.50) with CF. Body image perceptions in boys and girls with CF were examined in relation to the healthy control group using 2 (CF and control groups) by 2 (male and female) analysis of variance. The interaction effect was examined to explore the prediction that girls compared with boys with CF would have greater acceptance of their body shape and less desire to become larger. There were no differences between groups or sex in body esteem. On the CBIS body dissatisfaction score, children with CF were significantly more likely to perceive their ideal body size as a little larger than their current size while control children desired a smaller body size than their current size. CF children had a significantly lower mean score for body size satisfaction (an item assessing perception being too thin) and a significantly higher mean score on body weight satisfaction. There was a significant main effect of gender for only 1 measure, difference between the CBIS body dissatisfaction score, with girls being more likely to nominate a smaller ideal than their current figure. There were no significant interaction effects. Of children with CF and a low BMI (60% of amylase activity was consistent with the presence of MA. Immunoglobulin G (IgG) and immunoglobulin A (IgA) circulating autoantibodies to amylase were measured using recently developed enzyme-linked immunosorbent assay (ELISA), using porcine amylase as antigen. Results were expressed as arbitrary units (AUs). Statistical analysis was performed by Student's t test for unpaired data. IgA and IgG antibodies to exocrine pancreas tissue were detected by indirect immunofluorescence on human pancreas cryosections. RESULTS: Serum immunoprecipitation with either protein A Sepharose or polyethylene glycol reduced amylase activity from 1698 to 89 U/L (94.8%) and to 75 U/L (95.6%), with only marginal reduction in control serum samples. The ELISA for autoantibodies to amylase detected high values, both IgA (3531 AU) and IgG (1855 AU), in the serum sample from the patient at CD diagnosis. IgA autoantibodies (mean +/- standard deviation) were 3.4 +/- 2.5 AU in healthy controls, and 2.1 +/- 1.2 AU in celiac controls; IgG autoantibodies were 10 +/- 4.8 AU in healthy controls and 8.5 +/- 3.2 AU, respectively. Autoantibodies to exocrine pancreas tissue were documented in patient sera at the time of CD diagnosis, both IgA and IgG, but not in control groups. Preincubation of patient's serum with excess of alpha-amylase specifically inhibited antibody binding to coated amylase in the ELISA, and partially inhibited immunoreactivity to exocrine pancreas. Autoantibodies to alpha-amylase and to exocrine pancreas declined in CD patients after institution of a gluten-free diet. CONCLUSIONS: Few cases of MA have been described in children, and in all amylase determination was part of the clinical investigation for abdominal pain or trauma. (ABSTRACT TRUNCATED) PMID- 11389292 TI - Impact of breast pumping on lactogenesis stage II after cesarean delivery: a randomized clinical trial. AB - OBJECTIVE: Women at risk for delayed onset of lactation are often advised to pump their breasts before lactogenesis stage II to hasten the timing of this process. The effectiveness of this clinical practice has not been previously evaluated. This study investigates the effects of breast pumping before the onset of lactation on early milk transfer and subsequent breastfeeding duration among women giving birth by cesarean delivery. METHODOLOGY: Sixty women were randomly assigned to either the pumping group (n = 30), which used a double electric breast pump for six 10- to 15-minute sessions from 24 to 72 hours postpartum, or to the control group (n = 30), which held the pump to their breasts without suction for the same amount of time. Milk transfer was assessed by test weighing infants before and after 3 breastfeeding sessions daily. Test weight data were fitted to a second-order polynomial curve, to predict milk transfer over time. RESULTS: Breast pumping between 24 and 72 hours after cesarean delivery did not improve milk transfer. Participants in the pumping group tended to have lower milk transfer than did controls. Primiparae in the pumping group breastfed for ~5 months less than their counterparts in the control group; however, this difference was not statistically significant. CONCLUSIONS: Breast pumping did not improve milk transfer during the first 72 hours postpartum and may negatively affect breastfeeding duration among primiparous women. lactation, lactogenesis, breast milk, breast pumping, milk expression, breastfeeding, cesarean delivery. PMID- 11389293 TI - Diet quality, nutrient intake, weight status, and feeding environments of girls meeting or exceeding recommendations for total dietary fat of the American Academy of Pediatrics. AB - OBJECTIVES: To compare the diet quality and weight status of girls consuming diets meeting the recommendation of the American Academy of Pediatrics for dietary fat with those of girls consuming >30% of energy from fat and to examine relationships between girls' dietary fat intake, mothers' nutrient intakes, and mothers' child-feeding practices. DESIGN: Participants were 192 white girls and their mothers, who were divided into 2 groups: >30% of energy from fat (high fat [HF]) or /=5). Interventions. After cool humidified oxygen and 0.6 mg/kg of intramuscular dexamethasone, patients were randomized to receive either Heliox or RE. Vital signs, oxygen saturation, and CSs were recorded at regular intervals. OUTCOME/ANALYSIS: Reductions in CSs were compared using repeated measures analysis of variance. RESULTS: Thirty-three patients were enrolled. Three were excluded because of protocol violations, and 1 was excluded because of lack of documentation, leaving 29 patients for final analysis. The average age was 24.2 months, 20 were male (68.8%). Both Heliox and RE were associated with improvement in CSs over time. There were no significant differences in mean CS, oxygen saturation, respiratory rate, or heart rate between groups at baseline or at the end of the treatment period. CONCLUSION: In patients with moderate to severe croup, the administration of Heliox resulted in similar improvements in CS compared with patients given RE. PMID- 11389295 TI - Intussusception among recipients of rotavirus vaccine: reports to the vaccine adverse event reporting system. AB - BACKGROUND: Rotavirus vaccine was licensed on August 31, 1998, and subsequently recommended for routine use among infants. To assess rare adverse events, postlicensure surveillance was conducted. OBJECTIVE: To describe the cases of intussusception among rotavirus vaccine recipients reported to the Vaccine Adverse Event Reporting System from October 1998 through December 1999. SETTING AND PARTICIPANTS: Infants vaccinated with rotavirus vaccine in the United States. OUTCOME MEASURES: Intussusception confirmed by radiology, surgery, or autopsy report with medical record documentation or confirmed by a primary health care provider. RESULTS: There were 98 confirmed cases of intussusception after vaccination with rotavirus vaccine reported to the Vaccine Adverse Event Reporting System; 60 of these developed intussusception within 1 week after vaccination. Based on calculations using vaccine distribution data and intussusception incidence rates from 2 separate databases, an estimated 7 to 16 cases would have been expected to occur in the week after vaccination by chance alone. CONCLUSION: Using a passive surveillance system for vaccine adverse events, we observed at least a fourfold increase over the expected number of intussusception cases occurring within 1 week of receipt of rotavirus vaccine. Other studies were initiated to further define the relationship between rotavirus vaccine and intussusception. In light of these and other data, the rotavirus vaccine manufacturer voluntarily removed its product from the market, and the recommendation for routine use of rotavirus vaccine among US infants has been withdrawn. PMID- 11389296 TI - Contribution of residential exposures to asthma in us children and adolescents. AB - CONTEXT: Residential exposures are recognized risk factors for asthma, but the relative contribution of specific indoor allergens and their overall contribution to asthma among older children and adolescents in the United States are unknown. OBJECTIVE: To estimate the relative contributions, population-attributable risks, and costs of residential risk factors for doctor-diagnosed asthma. Design. Nationally representative, cross-sectional survey conducted from 1988 to 1994. SETTING AND PARTICIPANTS: A total of 5384 children who were 6 to 16 years old and participated in the National Health and Nutrition Examination Survey III, a survey of the health and nutritional status of children and adults in the United States. MAIN OUTCOME MEASURE: Doctor-diagnosed asthma, as reported by the parent. RESULTS: Five hundred three of 5384 children and adolescents (11.4%) had doctor diagnosed asthma. After adjusting for age, gender, race, urban status, region of country, educational attainment of the head of household, and poverty, predictors of doctor-diagnosed asthma included a history of allergy to a pet (odds ratio [OR: 2.4; 95% confidence interval [CI]: 1.7, 3.3), presence of a pet in the household (OR: 1.5; 95% CI: 1.1, 2.1), and immediate hypersensitivity to dust mite (OR: 1.5; 95% CI: 1.05, 2.0), Alternaria (OR: 1.9; 95% CI: 1.3, 2.8), and cockroach allergens (OR: 1.4; CI: 1.04, 1.9). Family history of atopy (OR: 1.7; 95% CI: 1.1, 2.7) and diagnosis of allergic rhinitis (OR: 2.1; CI: 1.1, 3.7) were also predictors for asthma. The population-attributable risk of having 1 or more residential exposures associated with doctor-diagnosed asthma was 44.4% (95% CI: 29-60), or an estimated 2 million excess cases. The attributable cost of asthma resulting from residential exposures was $405 million (95% CI: $264-$547 million) annually. CONCLUSIONS: The elimination of identified residential exposures, if causally associated with asthma, would result in a 44% decline in doctor diagnosed asthma among older children and adolescents in the United States. PMID- 11389297 TI - Deaths attributed to pediatric complex chronic conditions: national trends and implications for supportive care services. AB - BACKGROUND: Children with complex chronic conditions (CCCs) might benefit from pediatric supportive care services, such as home nursing, palliative care, or hospice, especially those children whose conditions are severe enough to cause death. We do not know, however, the extent of this population or how it is changing over time. OBJECTIVES: To identify trends over the past 2 decades in the pattern of deaths attributable to pediatric CCCs, examining counts and rates of CCC-attributed deaths by cause and age (infancy: <1 year old, childhood: 1-9 years old, adolescence or young adulthood: 10-24 years old) at the time of death, and to determine the average number of children living within the last 6 months of their lives. DESIGN/METHODS: We conducted a retrospective cohort study using national death certificate data and census estimates from the National Center for Health Statistics. Participants included all people 0 to 24 years old in the United States from 1979 to 1997. CCCs comprised a broad array of International Classification of Diseases, Ninth Revision codes for cardiac, malignancy, neuromuscular, respiratory, renal, gastrointestinal, immunodeficiency, metabolic, genetic, and other congenital anomalies. Trends of counts and rates were tested using negative binomial regression. RESULTS: Of the 1.75 million deaths that occurred in 0- to 24-year-olds from 1979 to 1997, 5% were attributed to cancer CCCs, 16% to noncancer CCCs, 43% to injuries, and 37% to all other causes of death. Overall, both counts and rates of CCC-attributed deaths have trended downward, with declines more pronounced and statistically significant for noncancer CCCs among infants and children, and for cancer CCCs among children, adolescents, and young adults. In 1997, deaths attributed to all CCCs accounted for 7242 infant deaths, 2835 childhood deaths, and 5109 adolescent deaths. Again, in 1997, the average numbers of children alive who would die because of a CCC within the ensuing 6-month period were 1097 infants, 1414 children, and 2548 adolescents or young adults. CONCLUSIONS: Population-based planning of pediatric supportive care services should use measures that best inform our need to provide care for time-limited events (perideath or bereavement care) versus care for ongoing needs (home nursing or hospice). Pediatric supportive care services will need to serve patients with a broad range of CCCs from infancy into adulthood. PMID- 11389299 TI - Vocal cord paralysis secondary to impacted esophageal foreign bodies in young children. AB - Impacted foreign bodies in the esophagus can result in respiratory symptoms including stridor and aphonia. Several mechanisms have been proposed to explain these symptoms, but the possibility of vocal cord paralysis and its cause has not been adequately emphasized. Two cases of young children with esophageal foreign body are described; both presented with respiratory symptoms, 1 with aphonia and the other with stridor. In both cases, the symptoms were secondary to vocal cord paralysis. A possible mechanism of recurrent nerve injury is proposed. PMID- 11389298 TI - Risk factors for improper vaccine storage and handling in private provider offices. AB - CONTEXT: Preventing loss of vaccine potency during storage and handling is increasingly important as new, more expensive vaccines are introduced, in at least 1 case requiring a different approach to storage. Little information is available about the extent to which staff in private physicians' offices meet quality assurance needs for vaccines or have the necessary equipment. Although the National Immunization Program at the Centers for Disease Control and Prevention (CDC) in 1997 developed a draft manual to promote reliable vaccine storage and to supplement published information already available from the CDC and the American Academy of Pediatrics, the best ways to improve vaccine storage and handling have not been defined. OBJECTIVES: To estimate the statewide prevalence of offices with suboptimal storage and handling, to identify the risk factors for suboptimal situations in the offices of private physicians, and to evaluate whether the distribution of a new National Immunization Program draft manual improved storage and handling practices. DESIGN: Population-based survey, including site visits to a stratified, random sample of consenting private physicians' offices. At least 2 months before the site visits, nearly half (intervention group) of the offices were randomly selected to receive a draft CDC manual entitled, "Guideline for Vaccine Storage and Handling." The remainder was considered the control group. Trained graduate students conducted site visits, all being blinded to whether offices were in the intervention or control groups. Each site visit included measurements of refrigerator and freezer temperatures with digital thermometers (Digi-thermo, Model 15-077-8B, Control Company, Friendswood, TX; specified accuracy +/- 1 degrees C). Their metal-tipped probes were left in the center shelf of cold storage compartments for at least 20 minutes to allow them to stabilize. The type of refrigerator/freezer unit, temperature-monitoring equipment, and records were noted, as were the locations of vaccines in refrigerator and freezer, and the presence of expired vaccines. Other information collected included the following: staff training, use of written guidelines, receipt of vaccine deliveries, management of problems, number of patients, type of office, type of medical specialty, and the professional educational level of the individual designated as vaccine coordinator. PARTICIPANTS: Two hundred twenty-one private physicians' offices known by the Georgia Immunization Program in 1997 to immunize children routinely with government-provided vaccines. OUTCOME MEASURES: Estimates (prevalence, 95% confidence interval [CI]) of immunization sites found to have a suboptimally stored vaccine at a single point in time, defined as: vaccine past expiration date, at a temperature of /=9 degrees C in a refrigerator or >/=-14 degrees C (recommended for varicella vaccine) in freezer, and odds ratios (ORs) for risk factors associated with outcomes. We performed chi(2) analysis and Student's t tests to compare the administrative characteristics and quality assurance practices of offices with optimal vaccine storage with those with suboptimal storage, and to compare the proportion of offices with suboptimal storage practices in the groups that did and did not receive the CDC manual. RESULTS: Statewide estimates of offices with at least 1 type of suboptimal vaccine storage included: freezer temperatures measuring >/=-14 degrees C = 17% (95% CI: 10.98, 23.06); offices with refrigerator temperatures >/=9 degrees C = 4.5% (95% CI: 1.08, 7.86); offices with expired vaccines = 9% (95% CI: 4.51, 13.37); and offices with at least 1 documented storage problem, 44% (95% CI: 35.79, 51.23). Major risk factors associated with vaccine storage outside recommended temperature ranges were: lack of thermometer in freezer (OR: 7.15; 95% CI: 3.46, 14.60); use of freezer compartment in small cold storage units (OR: 5.46; 95% CI = 2.70, 10.99); lack of thermometer in refrigerator (OR: 3.07; 95% CI: 1.15,8.20); and failure to maintain temperature log of freezer (OR: 2.70; 95% CI: 1.40, 5.23). Offices that adhered to daily temperature monitoring for all vaccine cold storage compartments, compared with those that did not, were 2 to 3 times more likely to assign this task to staff with higher levels of training, have received a recent visit from the state immunization program, and be affiliated with a hospital or have Federally Qualified Health Center status. In addition, sites using >1 refrigerator/freezer for vaccine storage were more likely to have at least 1 cold storage compartment outside recommended temperature ranges. We found no significant differences in the data reported above between the intervention group (received copy of the draft manual) and the control group (did not receive copy of draft manual), even when controlling for the annual number of immunizations given or the type of office. (ABSTRACT TRUNCATED) PMID- 11389300 TI - Diabetic autoimmune markers in children and adolescents with type 2 diabetes. AB - BACKGROUND: There is an increase in the incidence of type 2 diabetes in children and adolescents. Absence of known diabetes autoimmune markers is sometimes required to confirm the diagnosis. OBJECTIVE: To identify clinical and autoimmune characteristics of type 2 diabetes in a pediatric population. METHOD: We report an analysis of 48 children and adolescents with type 2 diabetes, compared with 39 randomly selected children with type 1 diabetes, diagnosed and followed at the Loma Linda University Pediatric Diabetes Center. Ethnic, familial, seasonal, and autoimmune marker characteristics are outlined. To determine the reliability of antibody testing in confirming the type of diabetes at diagnosis, we studied the incidence of positive islet cell antibodies (ICAs), glutamic acid decarboxylase antibodies (GADs), and insulin autoantibodies (IAAs) at diagnosis in both groups. ICA512, GADs, and IAAs were measured by radioimmunoassay. RESULTS: The cohort with type 2 diabetes had a similar gender distribution as the group with type 1 diabetes but a significantly higher age at diagnosis. Ethnic background was significantly different between the 2 groups, predominantly Hispanic in type 2 and white in type 1. Body mass index was significantly higher in type 2 diabetes (mean = 31.24 kg/m(2)). Among the patients with type 2 diabetes, 33% presented in diabetic ketoacidosis, random blood glucose at diagnosis ranged from 11.4 to 22.25 mmol/L (228-445 mg/dL), fasting C-peptide levels ranged from 0.89 to 2.7 nmol/L (2.7-8.2 ng/mL; normal: <1.36 nmol/L), and hemoglobin A(1C) was 10.8 +/- 3.5% (normal: <6.6%). None of these parameters was significantly different from the type 1 diabetes group. Although the incidence of diabetes antibody markers was significantly lower in type 2 versus type 1 diabetes, 8.1% of patients with type 2 diabetes had positive ICAs, 30.3% had positive GADs, and 34.8% had positive IAAs without ever being treated with insulin. In the type 2 diabetes group, none of the Hispanic patients had ICAs. However, there was no significant correlation between any of the diabetes antibodies and obesity, presence of acanthosis nigricans, or family history of diabetes. The frequency of thyroid antibodies was not significantly different from the group with type 1 diabetes. Daily insulin requirements 1 year after diagnosis were significantly lower in type 2 diabetes, ranging from 0 to 1.2 U/kg with a mean of 0.33. CONCLUSION: Absence of diabetes autoimmune markers is not a prerequisite for the diagnosis of type 2 diabetes in children and adolescents. PMID- 11389301 TI - Detection of neonatal carnitine palmitoyltransferase II deficiency by expanded newborn screening with tandem mass spectrometry. AB - The introduction of tandem mass spectrometry to newborn screening has substantially expanded our ability to diagnose metabolic diseases in the newborn period. We report the first case of neonatal carnitine palmitoyltransferase deficiency II detected by expanded newborn screening with tandem mass spectrometry. The neonate presented with dysmorphic facial features, structural malformations, renal failure, seizures, and cardiac arrythmias and died on the third day of life. This experience illustrates the importance of expanded newborn screening to avoid missing a metabolic diagnosis in early infantile death. PMID- 11389302 TI - Tunneled femoral central venous catheters in children with cancer. AB - OBJECTIVE: We discuss the feasibility of long-term femoral venous access by means of a cuffed subcutaneously tunneled central venous catheter (Broviac catheter) in selected pediatric cancer and stem cell transplant patients in whom access via the veins of the upper part of the torso is difficult or contraindicated and in whom alternative routes must be used. PATIENTS AND METHODS: We report on our experience with 9 patients (3 of whom underwent stem cell transplantation) who received femoral Broviac catheters between December 1990 and November 1999. Results. Time in place ranged from 4 to 155 days with a median of 58 days (mean: 71.2 days). Three catheters had to be removed: 1 because of infection of the subcutaneous tunnel and 2 because of catheter obstruction. The remaining 6 catheters functioned well without problems as long as they were needed; 1 of them got accidentally dislodged while the patient was off treatment. No episodes of catheter-related septicemia, thrombosis, kinking, or drug extravasation were noted; there were no catheter-related infectious complications in the transplant patients. CONCLUSIONS: Our experience indicates that in those instances in which customary access to the superior vena cava is precluded, long-term venous access by way of the femoral vein is a feasible and safe alternative in children, even in the setting of stem cell transplantation. PMID- 11389304 TI - Technical report: lawn mower-related injuries to children. AB - In the United States, approximately 9400 children younger than 18 years receive emergency treatment annually for lawn mower-related injuries. More than 7% of these children require hospitalization, and power mowers cause a large proportion of the amputations during childhood. Prevention of lawn mower-related injuries can be achieved by design changes of lawn mowers, guidelines for mower operation, and education of parents, child caregivers, and children. Pediatricians have an important role as advocates and educators to promote the prevention of these injuries. PMID- 11389303 TI - Once-a-day Concerta methylphenidate versus three-times-daily methylphenidate in laboratory and natural settings. AB - OBJECTIVE: Methylphenidate (MPH), the most commonly prescribed drug for attention deficit/hyperactivity disorder (ADHD), has a short half-life, which necessitates multiple daily doses. The need for multiple doses produces problems with medication administration during school and after-school hours, and therefore with compliance. Previous long-acting stimulants and preparations have shown effects equivalent to twice-daily dosing of MPH. This study tests the efficacy and duration of action, in natural and laboratory settings, of an extended release MPH preparation designed to last 12 hours and therefore be equivalent to 3-times-daily dosing. METHODS: Sixty-eight children with ADHD, 6 to 12 years old, participated in a within-subject, double-blind comparison of placebo, immediate release (IR) MPH 3 times a day (tid), and Concerta, a once-daily MPH formulation. Three dosing levels of medication were used: 5 mg IR MPH tid/18 mg Concerta once a day (qd); 10 mg IR MPH tid/36 mg Concerta qd; and 15 mg IR MPH tid/54 mg Concerta qd. All children were currently medicated with MPH at enrollment, and each child's dose level was based on that child's MPH dosing before the study. The doses of Concerta were selected to be comparable to the daily doses of MPH that each child received. To achieve the ascending rate of MPH delivery determined by initial investigations to provide the necessary continuous coverage, Concerta doses were 20% higher on a daily basis than a comparable tid regimen of IR MPH. Children received each medication condition for 7 days. The investigation was conducted in the context of a background clinical behavioral intervention in both the natural environment and the laboratory setting. Parents received behavioral parent training and teachers were taught to establish a school-home daily report card (DRC). A DRC is a list of individual target behaviors that represent a child's most salient areas of impairment. Teachers set daily goals for each child's impairment targets, and parents provided rewards at home for goal attainment. Each weekday, teachers completed the DRC, and it was used as a dependent measure of individualized medication response. Teachers and parents also completed weekly standardized ratings of behavior and treatment effectiveness. To evaluate the time course of medication effects, children spent 12 hours in a laboratory setting on Saturdays and medication effects were measured using procedures and methods adapted from our summer treatment program. Measures of classroom behavior and academic productivity/accuracy were taken in a laboratory classroom setting during which children completed independent math and reading worksheets. Measures of social behavior were taken in structured, small group board game settings and unstructured recess settings. Measures included behavior frequency counts, academic problems completed and accuracy, independent observations, teacher and counselor ratings, and individualized behavioral target goals. Reports of adverse events, sleep quality, and appetite were collected. RESULTS: On virtually all measures in all settings, both drug conditions were significantly different from placebo, and the 2 drugs were not different from each other. In children's regular school settings, both medications improved behavior as measured by teacher ratings and individualized target behaviors (the DRC); these effects were seen into the evening as measured by parent ratings. In the laboratory setting, effects of Concerta were equivalent to tid MPH and lasted at least through 12 hours after dosing. Concerta was significantly superior to tid MPH on 2 parent rating scores, and when asked, more parents preferred Concerta than preferred tid IR MPH or placebo. Side effects on children's sleep and appetite were similar for the 2 preparations. In the lab setting, both medications improved productivity and accuracy on arithmetic seatwork assignments, disruptive and on-task behavior, and classroom rule following. Both medications improved children's rule following and negative behavior in small group board games, as well as in unstructured recess settings. Individual target behaviors also showed significant improvement with medication across domains in the laboratory setting. Children's behavior across settings deteriorated across the laboratory day, and the primary effect of medication was to prevent this deterioration as the day wore on. Results support the use of background behavioral treatment in clinical trials of stimulant medication, and illustrate the utility of a measure of individualized daily target goals (ie, the DRC) as an objective measure of medication response in both the laboratory and natural school settings. CONCLUSION: This investigation clearly supports the efficacy of the Concerta long-acting formulation of MPH for parents who desire to have medication benefits for their child throughout the day and early evening. (ABSTRACT TRUNCATED) PMID- 11389305 TI - Managing stimulant medication for attention-deficit/hyperactivity disorder. PMID- 11389306 TI - Otitis externa. PMID- 11389307 TI - Preparticipation examination of the adolescent athlete: part 1. PMID- 11389308 TI - Vesicoureteral reflux. PMID- 11389309 TI - Index of suspicion. Case 1. Diagnosis: infected urachal cysts. PMID- 11389310 TI - Multidisciplinary teamwork: the good, bad, and everything in between. PMID- 11389311 TI - Choosing effective strategies for quality improvement. PMID- 11389312 TI - Accountability and quality improvement: the role of report cards. PMID- 11389313 TI - Erring is human: will we cross the quality chasm? PMID- 11389314 TI - Are members of multidisciplinary teams in breast cancer aware of each other's informational roles? AB - AIM: To conduct a commissioned survey of multidisciplinary breast team members' expectations of their own and each other's roles in providing different kinds of information to women with breast cancer. DESIGN: Questionnaire based survey. SETTING AND PARTICIPANTS: Health professionals from five multidisciplinary breast care centres within a Sussex health authority. MAIN OUTCOME MEASURES: Interdisciplinary awareness of informational roles played by different team members. RESULTS AND CONCLUSIONS: The results of the team survey suggest that, in most cases, health professionals fulfilled the roles expected of them by the team, with two or three individuals identified as the main providers of information for each topic. However, many more professionals were involved in major discussions without the team's knowledge. The professional consistently playing a major "unseen" role was the breast nurse specialist. PMID- 11389316 TI - Access to the evidence base from general practice: a survey of general practice staff in Northern and Yorkshire Region. AB - AIM: To identify and describe current methods of making health related research evidence accessible to general practice staff in the Northern and Yorkshire Region. METHOD: A postal survey questionnaire of general practice staff in the Northern and Yorkshire Region. RESULTS: At least one completed questionnaire was obtained from 70% of the general practices surveyed, and the individual response rate to the survey was 45%. Just under 60% of all respondents reported having no access to the NHS internet and just under 50% also reported having no access to the internet. All respondents in this survey reported greater access to paper based information than to electronic databases. However, this research provides evidence of differential access to information resources between different professions in general practice with GPs clearly having easier access than other professions to both paper based resources and electronic databases. 70% of all respondents said that they would need to be trained to use either a computer, the internet, or to search databases if the opportunity for easy access to any of these information services was available. CONCLUSIONS: At the time of this survey, general practices seemed to be struggling to set up the infrastructure and develop the skills that are necessary to make best use of available research evidence. In addition, there is a need for further investigation into the reasons why different professions working in the same practice setting have differential access to information resources available in primary care. PMID- 11389315 TI - Organisational strategies for changing clinical practice: how trusts are meeting the challenges of clinical governance. AB - OBJECTIVES: To describe the use, perceived effectiveness, and predicted future use of organisational strategies for influencing clinicians' behaviour in the approach of NHS trusts to clinical governance, and to ascertain the perceived benefits of clinical governance and the barriers to change. DESIGN AND SETTING: Whole population postal survey conducted between March and June 1999. SUBJECTS: Clinical governance leads of 86 NHS trusts across the South West and West Midlands regions. METHOD: A combination of open questions to assess the use of strategies to influence clinician behaviour and the barriers to clinical governance. Closed (yes/no) and Likert type ratings were used to assess the use, perceived effectiveness, and future use of 13 strategies and the predicted outcomes of clinical governance. RESULTS: All trusts use one or more of 13 strategies categorised as educational, facilitative, performance management, and organisational change methods. Most popular were educational programmes (96%) and protocols and guidelines (97%). The least popular was performance management such as use of financial incentives (29%). Examples of successful existing practice to date showed a preference for initiatives that described the use of protocols and guidelines, and use of benchmarking data. Strategies most frequently rated as effective were facilitative methods such as the facilitation of best practice in clinical teams (79%), the use of pilot projects (73%), and protocols and guidelines (52%). The least often cited as effective were educational programmes (42%) and training clinicians in information management (20%); 8% found none of the 13 strategies to be effective. Predicted future use showed that all the trusts which completed this section intended to use at least one of the 13 strategies. The most popular strategies were educational and facilitative. Scatterplots show that there is a consistent relationship between use and planned future use. This was less apparent for the relationship between planned use and perceived effectiveness. Barriers to change included lack of resources, mainly of money and staff time, and the need to address cultural issues, plus infrastructure support. The anticipated outcomes of clinical governance show that most trusts expect to influence clinician behaviour by improving patient outcomes (78%), but only 53% expect it to result in better use of resources, improved patient satisfaction (36%), and reduced complaints (10%). CONCLUSIONS: Clinical governance leads of trusts report using a range of strategies for influencing clinician behaviour and plan to use a similar range in the future. The choice of methods seems to be related to past experience of local use, despite equivocal judgements of their perceived effectiveness in the trusts. Most expect to achieve a positive impact on patient outcomes as a result. It is concluded that trusts should establish methods of learning what strategies are effective from their own data and from external comparison. PMID- 11389317 TI - Age, gender, socioeconomic, and ethnic differences in patients' assessments of primary health care. AB - BACKGROUND: Patients' evaluations are an important means of measuring aspects of primary care quality such as communication and interpersonal care. This study aims to examine variations in assessments of primary care according to age, gender, socioeconomic, and ethnicity variables. METHODS: A cross sectional survey of consecutive patients attending 55 inner London practices was performed over a 2 week period using the General Practice Assessment Survey (GPAS) instrument which assesses 13 important dimensions of primary care provision. Variations in scale scores were investigated for differences relating to age, gender, socioeconomic, and ethnic status as reported by respondents. RESULTS: A total of 7692 questionnaires were returned (71% response rate). Valid information on age, gender, socioeconomic status, and ethnicity was available for 4819 out of 5496 adult respondents. Approximately half the respondents reported their ethnic group as "white" and most of the remaining respondents reported belonging to "black" or South Asian groups. Significant differences existed between groups of patients defined by age or ethnicity for most of the scale scores examined. Black, South Asian, and Chinese respondents reported lower scores (representing less favourable assessments) than white respondents; older respondents reported more favourable evaluations of care than younger respondents; and less affluent groups reported lower scores than more affluent groups for two of the 13 dimensions. There was no significant difference between gender groups with respect to assessment of primary care. Age and ethnicity were independent predictors of respondents' assessments of primary care. CONCLUSIONS: Differences exist between identifiable subgroups of the population in their assessments of primary health care measured using the GPAS instrument. This work adds to the literature on variation in healthcare experience and the potential for patient assessment of primary care. Further work is required to investigate these differences in more detail and to relate them to differences in the nature and process of primary care provision. Primary care providers need to ensure that services provided are appropriate for all patient groups within their communities. PMID- 11389318 TI - Publicly disclosed information about the quality of health care: response of the US public. AB - Public disclosure of information about the quality of health plans, hospitals, and doctors continues to be controversial. The US experience of the past decade suggests that sophisticated quality measures and reporting systems that disclose information on quality have improved the process and outcomes of care in limited ways in some settings, but these efforts have not led to the "consumer choice" market envisaged. Important reasons for this failure include limited salience of objective measures to consumers, the complexity of the task of interpretation, and insufficient use of quality results by organised purchasers and insurers to inform contracting and pricing decisions. Nevertheless, public disclosure may motivate quality managers and providers to undertake changes that improve the delivery of care. Efforts to measure and report information about quality should remain public, but may be most effective if they are targeted to the needs of institutional and individual providers of care. PMID- 11389319 TI - Public release of performance data and quality improvement: internal responses to external data by US health care providers. AB - Health policy in many countries emphasises the public release of comparative data on clinical performance as one way of improving the quality of health care. Evidence to date is that it is health care providers (hospitals and the staff within them) that are most likely to respond to such data, yet little is known about how health care providers view and use these data. Case studies of six US hospitals were studied (two academic medical centres, two private not-for-profit medical centres, a group model health maintenance organisation hospital, and an inner city public provider "safety net" hospital) using semi-structured interviews followed by a broad thematic analysis located within an interpretive paradigm. Within these settings, 35 interviews were held with 31 individuals (chief executive officer, chief of staff, chief of cardiology, senior nurse, senior quality managers, and front line staff). The results showed that key stakeholders in these providers were often (but not always) antipathetic towards publicly released comparative data. Such data were seen as lacking in legitimacy and their meanings were disputed. Nonetheless, the public nature of these data did lead to some actions in response, more so when the data showed that local performance was poor. There was little integration between internal and external data systems. These findings suggest that the public release of comparative data may help to ensure that greater attention is paid to the quality agenda within health care providers, but greater efforts are needed both to develop internal systems of quality improvement and to integrate these more effectively with external data systems. PMID- 11389320 TI - Accreditation and the quality movement in France. PMID- 11389321 TI - Promoting the initiation of breast feeding. PMID- 11389325 TI - Helicoductus thulakoceras n. g., n. sp. (Cestoda: Hymenolepididae) a parasite of Charadrius marginatus (Aves: Charadrii) from South Africa. AB - The authors describe and illustrate Helicoductus thulakoceras n. g., n. sp., a parasite of Charadrius marginatus (Aves: Charadrii) from South Africa. This hymenolepidid cestode is 1.5-3 mm long, with a rostellum armed with 10 diorchoid hooks 75-80 microm long (mean 77 microm) and anomalous terminal genital ducts. One spine (10-15 microm long) is inserted at the ventral extremity of the male genital pouch. An invaginated cirrus is absent. A non-retractable and non invaginable external tube, 25-30 x 4-2.5 microm, appears as a spiral, sclerotinised, spinous canal, which is non-retractable, non-invaginable and directed dorso-ventrally from the genital pouch to a second ventral canal parallel with the male genital pouch; the second canal is thin and spine-like (15 20 x 1.5 microm) with a thick hyaline muff. These peculiar anatomical structures are discussed. They are considered to justify the erection of a new genus. PMID- 11389326 TI - Biogeography of helminth parasitism in Lemmus Link (Arvicolinae), with the description of Paranoplocephala fellmani n. sp. (Cestoda: Anoplocephalidae) from the Norwegian lemming L. lemmus (Linnaeus). AB - We describe the gastrointestinal helminth fauna of true lemmings (Lemmus spp., Arvicolinae) based on published and original material throughout the Holarctic range of these hosts. According to the existing data, the helminth fauna of true lemmings consists of three widespread and/or locally common taxa: Hymenolepis horrida (sensu lato) (Hymenolepididae), Anoplocephaloides lemmi (Anoplocephalidae) and Heligmosomoides spp. (Heligmosomidae). Despite the taxonomic boundaries and ancient phylogenetic splits in the hosts, there are no major faunistic differences for parasites within western (Siberian) L. sibiricus and L. bungei, and eastern (North American) L. trimucronatus throughout their distribution range. In contrast, the Norwegian lemming L. lemmus, which is a Fennoscandian endemic and closely related to the western populations of L. sibiricus, has only a single host-specific helminth, the cestode Paranoplocephala fellmani n. sp. (Anoplocephalidae). We describe the new species and show that it differs consistently from related species by its long and slender cirrus-sac. However, there are also a number of other significant differences, e.g., P. fellmani n. sp. and Andrya primordialis in Tamiasciurus hudsonicus (Sciuridae) evidently have a unique (sub)type of uterine development among Andrya/Paranoplocephala spp. Because P. fellmani n. sp. was also found to occur in Alaska (host L. trimucronatus), this species seems to follow the same biogeographical pattern as the other specialist helminths of Lemmus. We suggest alternative explanations for the absence of three major helminth taxa in the Norwegian lemming in Fennoscandia. PMID- 11389327 TI - New reports and a redescription of Porrocaecum heteropterum (Diesing, 1851) (Ascarididae), a rare nematode parasitic in South American.threskiornithid birds1. AB - Porrocaecum heteropterum (Diesing, 1851) (Nematoda, Ascarididae) is reported parasitising the white-faced ibis Plegadis chihi and the black-faced ibis Theristicus melanopis melanopis (Ciconiiformes, Threskiornithidae) from the Provinces of Buenos Aires and Neuquen, Argentina. This nematode has been reported very few times in the literature, mainly from Brazilian threskiornithids, and there have been no new reports following a redescription of the species given in 1957. This paper provides new host and locality records for this rather rare species, as well as some additional morphological data, mainly based on SEM studies, which complement the previous descriptions. The scarce and sporadic records of this species seem to indicate not only a defined host-specificity towards threskiornithid birds but also that the acquisition of this parasite is possible only when certain ecological barriers, including food availability, feeding habits and environmental conditions, are surmounted. PMID- 11389328 TI - Metacestodes of the family Dilepididae (Cestoda: Cyclophyllidea) parasitising fishes in Mexico. AB - A survey of metacestodes of dilepidid tapeworms (Cyclophyllidea) occurring in fish from Mexico is presented. They belong to the following species (those first reported from Mexico marked with an asterisk): Cyclustera capito (Rudolphi, 1819); (*)Cyclustera cf. ralli (Underwood & Dronen, 1986); Dendrouterina pilherodiae Mahon, 1956; (*)Glossocercus auritus (Rudolphi, 1819); (*)G. caribaensis (Rysavy & Macko, 1973); (*)Paradilepis caballeroi Rysavy & Macko, 1973; (*)Paradilepis cf. urceus (Wedl, 1855); (*) Paradilepis sp.; Parvitaenia cochlearii Coil, 1955; (*)Parvitaenia macropeos (Wedl, 1855); (*)Valipora campylancristrota (Wedl, 1855); (*)V. mutabilis Linton, 1927; and (*)V. minuta (Coil, 1950). Metacestodes of Dendrouterina papillifera (Fuhrmann, 1908), previously reported from the gall-bladder of the pimelodid catfish Rhamdia guatemalensis from Mexico by Scholz et al. (1996), belong actually to V. minuta. Data on the morphology of metacestodes, their fish hosts and rate of infection, site and distribution in Mexico are provided. PMID- 11389329 TI - Variation of cuticular characters in the Nematomorpha: studies on Gordionus violaceus (Baird, 1853) and G. wolterstorffii (Camerano, 1888) from Britain and Ireland. AB - Gordionus violaceus (Baird, 1853) and G. wolterstorffii (Camerano, 1888) are regarded as two species which are clearly separated by distinct cuticular patterns. A study of 59 specimens of these two species from Britain and Ireland revealed numerous transitional stages between the cuticular patterns that are regarded as being typical for each species. Variation was also found at the posterior end, but no character separates both species. Therefore, G. violaceus and G. wolterstorffii might represent one polymorphic species. Four specimens resembled G. violaceus but had a different cuticular pattern, including a second type of areole occurring in a pattern which has been named a megareolar pattern. This might represent a separate species, but a detailed determination relies upon further revisional work in the genera Gordionus, Parachordodes and Paragordionus. PMID- 11389330 TI - Biogeography and phylogeny of Dermoergasilus Ho & Do, 1982 (Copepoda: Ergasilidae), with descriptions of three new species. AB - Three new species of Dermoergasilus are described from six species of grey mullet hosts. Dermoergasilus longiabdominalis n. sp. was found on Valamugil engeli (Bleeker) from the Philippines and Madagascar and on V. cunnesius (Valenciennes) from the Philippines and Mangalore, India. D. semiamplectens n. sp. occurred on Sicamugil hamiltoni (Day) from the Sittang River, Burma, on Liza subviridis (Valenciennes) and L. parsia (Hamilton Buchanan) from Calcutta, India, and on V. cunnesius (Valenciennes) from China. D. curtus n. sp. parasitised Rhinomugil squamipinnis (Swainson) from Alahabad, India. A key to the ten currently accepted species of Dermoergasilus is given. The biogeographical distribution of Dermoergasilus species is analysed and levels of host-specificity are surveyed within the genus. The phylogenetic relationships between the species of Dermoergasilus are also analysed. PMID- 11389331 TI - Grillotia australis n. sp. and G. pristiophori n. sp. (Cestoda: Trypanorhyncha) from Australian elasmobranch and teleost fishes. AB - Two new species of Grillotia are described from elasmobranch and teleost fishes from south-eastern Australia. G. australis n. sp., from the Australian angel shark Squatina australis. Regan, most closely resembles G. smarisgora (Wagener, 1854) and G. angeli Dollfus, 1969, differing from both species in the presence of smaller bulbs, two or occasionally three hooks in each intercalary row in the basal region, reduced to one in the metabasal region compared with four or five hooks in the metabasal region of G. smarisgora and a single hook in G. angeli, and in the limited extent of the band of hooklets on the external surface in the basal region of the tentacle, a region which is covered with hooks in G. smarisgora. Plerocerci of this species were found in the mackerel Trachurus declivis (Jenys) (site not known) from Tasmania. G. pristiophori n. sp., from the saw sharks Pristiophorus cirratus (Latham) and P. nudipinnis Gunther, most closely resembles G. spinosissima Dollfus, 1969 in possessing a scolex covered with spiniform microtriches, but differs in having six rather than five hooks in each principal row, no intercalary hooks and by possessing a band of hooklets on the external surface of the tentacle which diminishes distally into a single file, rather than persisting as a band eight to nine files wide. G. pristiophori is the first trypanorhynch to be recorded from saw-sharks. PMID- 11389332 TI - Description of the male and first-stage juvenile of Longidorus intermedius Kozlowska & Seinhorst, 1979 (Nematoda: Dorylaimida), and notes on its morphology and distribution. AB - Longidorus intermedius Kozlowska & Seinhorst, 1979 is redescribed from specimens recovered from oak (Quercus L.) forests in Bulgaria. The male and the first-stage juvenile were recorded for the first time, as were populations from Macedonia and Turkey. L. intermedius is identified here as the first species in the genus with only three juvenile stages and a pan-European distribution. The species was recovered from 36 of 46 oak forests in Bulgaria, found associated with each of nine oak species sampled and reached population densities of up to 289 specimens per 200 cm(3) of soil. The distribution of L. intermedius in Bulgaria, and in other countries, suggests that this species, like Xenocriconemella macrodora (Taylor, 1936) and probably L. aetnaeus Lamberti, Agostinelli & Vinciguerra, 1986, can be used as an ecological indicator for habitats where Quercus associations represent a primary vegetation type. PMID- 11389333 TI - Two new species of Neolebouria Gibson, 1976 (Digenea: Opecoelidae) from temperate marine fishes of Australia. AB - Neolebouria moretonensis n. sp. is described from Gerres subfasciatus (Gerreidae) from Moreton Bay, south-east Queensland and N. lineatus n. sp. is described from Centroberyx lineatus (Berycidae) from off Rottnest Island, south-west Western Australia. C. lineatus represents a new host family and order (Beryciformes) for the genus. The two new species are distinguished within the genus by their entire, tandem to oblique testes and cirrus-sacs that do not extend into the hind body, by being less than 1 mm in length, and by the position of the genital pore and the relative size of the forebody and post-testicular region. N. lineatus and N. moretonensis are very similar but are distinguished by their caeca which extend further posteriorly in N. moretonensis. There is no apparent pattern in the host-specificity of this genus. PMID- 11389334 TI - New species of Hatschekia Poche, 1902 (Copepoda: Hatschekiidae) parasitic on marine fishes of Kuwait. AB - Three new species of Hatschekia are described from the gill filaments of fishes taken in the Persian Gulf: H. seyi n. sp. from Heniochus acuminatus (Linnaeus), H. nodosa n. sp. from Lutjanus coccineus (Cuvier) and H. tanysoma n. sp. from L. fulviflamma (Forsskal). H. seyi differs from its congeners in having the cephalothorax longer than the trunk; H. nodosa, in having multiple nodules on the posterolateral margins of the trunk; and H. tanysoma, in having the trunk more than six times the length of cephalothorax and some pinnate setae on legs 1 and 2. PMID- 11389335 TI - Bartolius pierrei n. g., n. sp. (Digenea: Gymnophallidae) from the Peninsula Valdes, Argentina. AB - Bartolius pierrei n. g., n. sp. (Digenea: Gymnophallidae) is described from metacercariae and naturally and cultivated obtained adults from southern Argentina. The second intermediate host is Darina solenoides (King) (Bivalvia: Mactridae) and the definitive host is Larus dominicanus Lichtenstein (Aves: Laridae). The diagnostic characters are as follows: Body small, oval. Oral sucker without lateral projections, twice size of ventral sucker (except in young metacercariae). Caeca short (in adults), without dorsal diverticula. Ventral sucker in posterior third of body. Ventral pit absent. Seminal vesicle bipartite. Ovary post-testicular. Vitelline glands paired, compact, close to ventral sucker. Uterus in fore- and hindbody. Genital atrium tubular. Genital pore inconspicuous, close to anterior margin of ventral sucker. Excretory vesicle Y-shaped with very short stem. Excretory formula: 2[(2+2)+(2+2)] = 16. Bartolius is distinguished from other genera of the Gymnophallidae by the post-testicular position of the ovary. PMID- 11389336 TI - A new species of Acusicola Cressey (Copepoda: Ergasilidae) from northeastern Brazil. AB - A new species of Acusicola is described based on adults of both sexes taken from plankton samples collected in the upper reaches of the Piaui River estuary, in the northeast of Brazil. Ovigerous females were present in the plankton. The new species, Acusicola minuta n. sp., can be distinguished from its congeners by its small body size, female antennal morphology and leg setation. The male described here as A. minuta n. sp. is the first known male attributed to the genus. PMID- 11389337 TI - The Calicotyle conundrum: do molecules reveal more than morphology? AB - Partial large subunit 28S rDNA sequences were obtained for specimens of Calicotyle (Monogenea: Monocotylidae) from eight different host species distributed worldwide to test the validity of some species and to address the question of host-specificity in others. Sequences obtained for Calicotyle specimens identified as C. kroyeri based on morphological methods from the type host Raja radiata (Rajidae) and an additional host R. clavata, both from the North Sea, were identical. However, 'C. kroyeri' from the cloaca of R. naevus from Tunisia, Raja sp. A from Tasmania and R. radula from Tunisia differed from C. kroyeri from R. radiata by five (0.51%), 21 (2.13%) and 39 (3.96%) base pairs, respectively, over 984 sites. Therefore, it is likely that the specimens from Raja sp. A, R. radula and perhaps even from R. naevus are not C. kroyeri. Molecular results determined that the calicotylines from the cloaca of Urolophus cruciatus and U. paucimaculatus (Urolophidae) from southern Tasmania identified previously as C. urolophi are indeed identical. Large subunit 28S rDNA sequences of C. palombi and C. stossichi collected from the cloaca and rectal gland, respectively of Mustelus mustelus (Triakidae) from the coast of Tunisia differ sufficiently for these calicotylines to be considered separate and valid species. Our results indicate that some species of Calicotyle are not strictly host specific, but that C. kroyeri may not be as widely distributed in rajids as was believed previously. Calicotyle specimens from rajids must be re-examined critically to determine whether there are morphological differences indicative of specific differences that may have been overlooked previously. PMID- 11389339 TI - Manifestations of the shaken baby syndrome. AB - Retinal hemorrhages are the most common fundus finding in the shaken baby syndrome. They vary in type and location; no particular type is pathognomonic for the condition. Retinal hemorrhages are not needed to make a diagnosis of shaken baby syndrome. However, in a child under age 3 years, the presence of extensive bilateral retinal hemorrhages raises a very strong possibility of abuse, which must be investigated. The other possible causes for hemorrhages in this age child can be investigated and eliminated. The diagnosis of abuse should be made by someone particularly trained in this area, who can put together the entire picture of inadequate or changing history, fractures of various ages, particularly rib fractures, subdural hematoma of the brain, and retinal hemorrhages. Photographs of retinal hemorrhages are very helpful to child advocacy experts who take these cases to court. PMID- 11389340 TI - Laser therapy for retinopathy of prematurity. AB - Retinopathy of prematurity (ROP) is a potentially blinding condition that afflicts preterm infants in the neonatal period. Although advances in scleral buckling and vitrectomy techniques offer hope for those infants suffering from stage 4 or 5 ROP, prevention of progression to these stages offers the most promise for favorable structural and visual outcomes. Proper screening for threshold ROP and treatment with peripheral retinal ablation are the keys to successfully managing ROP. Technological advances in screening tools and portable diode lasers enable ophthalmologists to provide prompt, effective, and safe treatment for patients with threshold ROP. PMID- 11389341 TI - Laser therapy for rhegmatogenous retinal detachment. AB - Traditional techniques for managing uncomplicated macula-sparing rhegmatogenous retinal detachments include scleral buckling and pneumatic retinopexy. Demarcation laser photocoagulation is associated with less morbidity than these techniques and may be equally as effective in stabilizing selected macula-sparing retinal detachments. PMID- 11389342 TI - Cataract surgery in diabetes. AB - Recent studies of cataract surgery in diabetes confirm retinopathy severity and macular edema as the principal determinants of postoperative visual acuity, and link improved visual outcomes to the shift from conservative management toward earlier surgical intervention. PMID- 11389343 TI - Suprachoroidal hemorrhage during pars plana vitrectomy. AB - Suprachoroidal hemorrhage is an uncommon but serious complication of pars plana vitrectomy that can be associated with a guarded visual prognosis. Risk factors for development of suprachoroidal hemorrhage during pars plana vitrectomy include high myopia, history of previous retinal detachment surgery, rhegmatogenous retinal detachment, use of cryotherapy, scleral buckling at the time of pars plana vitrectomy, external drainage of the subretinal fluid, intraoperative systemic hypertension, and bucking during general anesthesia. In eyes with suprachoroidal hemorrhage during pars plana vitrectomy, the final visual and anatomic outcomes may be compromised by persistent retinal detachment, secondary glaucoma, and ocular hypotony. In most cases, intraoperative drainage of suprachoroidal hemorrhage is not associated with a better outcome. The prognosis is more favorable if the suprachoroidal hemorrhage is localized and does not extend in to the posterior pole. PMID- 11389344 TI - Ocular paintball injuries. AB - Paintball sport-related ocular injuries represent an increasing problem as the popularity of the sport increases and the number of participants grows. Although eye protective devices designed specifically for paintball sports are extremely effective in preventing such injuries, the failure to properly wear these devices has resulted in an alarming number of severe ocular injuries. Recent trends have indicated that an increasing percentage of paintball sport-related ocular injuries have occurred in unsupervised, noncommercial settings (i.e., backyard games) where the use of eye protective devices is not required. Paintball industry standards for eye protection have recently been developed and should be implemented for all participants. PMID- 11389346 TI - Cataract surgery after retinal surgery. AB - Many patients require surgery for cataract after retinal surgery. When evaluating patients preoperatively, the surgeon should pay careful attention to the assessment of visual function and potential acuity, and to the slit lamp examination of the lens, posterior capsule, anterior cortical vitreous, and zonules. The surgeon's technique must account for increased nuclear sclerosis, intraoperative miosis, a characteristically longer axial length, and fluctuations in anterior chamber depth from movement of the iris-lens diaphragm. Posterior capsule plaques are not unusual. Posterior capsule opacification is the most frequent late complication. Visual outcomes are usually good except when limited by pre-existing macular pathology. Vision-threatening complications of cataract surgery are unusual. PMID- 11389345 TI - Viral causes of the acute retinal necrosis syndrome. AB - Acute retinal necrosis has been described as a clinical entity for nearly 30 years. Acute retinal necrosis is a potentially visually devastating necrotizing vaso-occlusive retinitis affecting both healthy and immunocompromised patients. Acute retinal necrosis is caused by the herpes group of viruses, mainly varicella zoster, herpes simplex types 1 and 2, and, rarely, cytomegalovirus. Recently, polymerase chain reaction techniques have enabled detection of very small amounts of viral DNA from intra-ocular fluid samples. This can help in both the diagnosis of atypical cases of retinitis and uveitis and directing treatment in cases of acute retinal necrosis. PMID- 11389347 TI - Photodynamic therapy update. AB - Photodynamic therapy uses a photoactivating agent to selectively treat choroidal neovascularization. In April 2000, the United States Food and Drug Administration approved verteporfin photodynamic therapy for the treatment of subfoveal, predominately classic, choroidal neovascularization caused by age-related macular degeneration. The treatment of choroidal neovascularization from other causes such as myopia, angioid streaks, and idiopathy, and presumed ocular histoplasmosis syndrome is still under investigation. Other photoactivating agents are being evaluated. Photodynamic therapy has been shown to halt the progression of visual loss in patients with age-related macular degeneration who have subfoveal predominately classic choroidal neovascularization. The socio economic impact of verteporfin approval has yet to be determined. PMID- 11389348 TI - Treatment of subretinal hemorrhages with tissue plasminogen activator. AB - Submacular hemorrhages pose a danger to visual acuity. Intervention may help prevent or mitigate severe visual loss. Pneumatic displacement and vitrectomy with direct evacuation are two methods of treating submacular hemorrhages. Tissue plasminogen activator may be an important adjunct to these techniques. PMID- 11389349 TI - Transpupillary thermotherapy of subfoveal occult choroidal neovascularization. AB - Choroidal neovascularization secondary to age-related macular degeneration is a leading cause of vision loss in adults. Although most patients present with occult CNV, treatment has focused on the small percentage of eyes with well delineated, classic CNV. Transpupillary thermotherapy is a recent advancement in the management of occult CNV. Transpupillary thermotherapy acts in a subthreshold manner by slightly raising the choroidal temperature. A recent pilot study demonstrated that 56% of treated eyes remained stable one year after treatment with only 25% losing two lines of visual acuity. The TTT4CNV study will further evaluate the effectiveness of transpupillary thermotherapy in a randomized, double-blind trial. PMID- 11389350 TI - Pars plana vitrectomy and sutured posterior chamber lens implantation. AB - Transcleral suturing of posterior chamber intraocular lenses (IOLs) was developed to extend the benefits of IOL visual rehabilitation to eyes lacking capsular support and to avoid the complications of closed loop interior chamber IOLs. Although most frequently implanted after complicated cataract surgery or penetrating keratoplasty, they are indicated in several situations following pars plana vitrectomy. The surgical techniques have evolved to minimize the risk of complications, but the surgery remains technically more demanding and time consuming than insertion of a modern open loop anterior chamber IOL. No randomized trials have compared the relative risks of each lens type, which leaves surgeon preference as the major determinant of what lens is implanted. It is likely, however, that in eyes with extensive anterior chamber angle damage or large iris defects, sutured posterior chamber IOLs will remain the first choice for surgical rehabilitation. PMID- 11389351 TI - Natural history of choroidal neovascularization in high myopia. AB - High myopia, or pathologic myopia, usually refers to a condition in which individuals have greater than 6 diopters of myopia or an axial length greater than 26 to 27 mm. The natural history of choroidal neovascularization in high myopia is variable, and reports to date have some conflicting information, but analysis shows fairly poor final visual outcomes. This outcome complicates recommendations for treatment and must be considered in future studies aimed at managing this disease. PMID- 11389352 TI - Treatment of Propionibacterium acnes endophthalmitis. AB - Propionibacterium acnes is a well-known cause of delayed endophthalmitis following cataract surgery. A white intracapsular plaque, keratic precipitates, and hypopyon may be present. Although there is no consensus on the treatment approach, recent studies have reviewed the outcomes of relatively large numbers of patients. Reasonable treatment options include pars plana vitrectomy and intra ocular antibiotics with either partial or total capsulectomy/lens exchange. A good visual outcome is possible. PMID- 11389353 TI - The cost-effectiveness of early vitrectomy for the treatment of vitreous hemorrhage in diabetic retinopathy. AB - Diabetic vitrectomy has been found to be efficacious for the treatment of vitreous hemorrhage secondary to diabetic retinopathy. The purpose of this study is to determine the cost-effectiveness of early vitrectomy for the management of vitreous hemorrhage secondary to diabetic retinopathy. The analysis was performed from the perspective of a third-party insurer. A cost-utility Markov model was used to determine the cost per quality-adjusted life year (QALY) gained from early versus deferral of vitrectomy. The model used 2-, 3-, and 4-year results from the Diabetic Retinopathy Vitrectomy Study, patient-based utilities, life expectancy data, and incremental medical costs. Early vitrectomy was the dominant strategy and was associated with a gain of 0.41 QALYs over the 57-year expected life span for a hypothetical patient. The cost per additional QALY gained from early vitrectomy treatment was $1910 (US$ discounted at 3%). When sensitivity analyses were performed by varying efficacy probabilities and utilities across their 95% confidence intervals, early treatment was always the dominant strategy. Additionally, even at the extreme sensitivity values, the cost per QALY of early vitrectomy treatment remained under $10,000. Overall, early vitrectomy for the treatment of vitreous hemorrhage secondary to diabetic retinopathy is highly cost effective. PMID- 11389354 TI - Rethinking the physiologic paradigms in the critically ill. PMID- 11389355 TI - Neonates with congenital heart disease. AB - Early reparative surgery in neonates and infants with congenital heart disease, as opposed to initial palliation and later repair, is now commonplace. Changes to the conduct of cardiopulmonary bypass, timing of surgery and surgical techniques, and perioperative management substantially have reduced the postoperative mortality and morbidity for these patients. The success of this strategy of early reparative surgery now has been extended to the premature and low-birth-weight newborn, and, along with this, new challenges to postoperative care in the intensive care unit. However, the low mortality associated with two-ventricle repairs has not been the experience in newborns undergoing palliation for single ventricle defects, in particular, hypoplastic left heart syndrome. A number of articles regarding management of newborns with single-ventricle defects have been published during the past 12 months, ranging from classification, prenatal diagnosis, treatment options, and predictors of both early and late outcome, which may provide a guide for patient management. As mortality has declined, there has been an increased emphasis on identifying indices that may predict outcome or morbidity both before and after surgery, along with possible strategies to attenuate adverse clinical responses. The inflammatory response to bypass is heightened in neonates and infants, and several reports have addressed possible techniques for attenuating the response. In addition, reports regarding the risk for necrotizing enterocolitis, the utility of lactate as an index of systemic perfusion, potential markers of myocardial and neurologic injury, and the use of mechanical support of the circulation in newborns with congenital heart disease are summarized. PMID- 11389356 TI - Neonatal brain injury. AB - Complacency about long-term outcomes in newborns is being eroded rapidly with new information. We have examined developments in the area from an explicitly clinical approach, focusing on etiology, diagnostic modalities, and therapies. We attempt to discuss relevance from the preterm and the term perspective. Emerging evidence implicating chorioamnionitis as a significant contributor to neonatal brain injury is discussed. Therapeutic modalities such as magnetic resonance imaging and electrophysiological monitoring offer some potentially new tools for the clinician. An exploding series of basic advances suggest several potentially new strands of therapy. We discuss two that deserve further clinical exploration, namely anti-inflammatory strategies and thread hormone supplementation. In the arena of therapy, however, the paucity of large trials from which to guide therapies is a predominant theme, leaving a large reservoir of uncertainty for the clinician. PMID- 11389357 TI - Acute myocarditis. AB - Myocarditis is defined as inflammation of the myocardium accompanied by myocellular necrosis. Acute myocarditis must be considered in patients who present with recent onset of cardiac failure or arrhythmia. Often there is a history of an antecedent flu-like illness. Fulminant myocarditis is a distinct entity characterized by sudden onset of severe congestive heart failure or cardiogenic shock, usually following a flu-like illness. Giant cell myocarditis is a rare, frequently fatal disorder of unknown origin characterized by presence of giant cell inflammatory infiltrate in the myocardium. In recent years we have made good progress in understanding the causes, pathogenesis, natural history, diagnosis, and treatment of myocarditis. However, our knowledge is still far from complete. New information that extends our understanding of myocarditis is being reported constantly. This review summarizes recent advances in myocarditis, with an emphasis on the literature during the last year. PMID- 11389358 TI - Nonpharmacologic treatment of acute heart failure. AB - Acute heart failure is unusual in the pediatric population, but in many situations it justifies aggressive therapy. For example, children with lymphocytic myocarditis have an overall survival rate of nearly 90%, with complete myocardial recovery for the majority. Pharmacologic agents traditionally have been the mainstay of medical therapy for acute heart failure, but, in recent years, there has been increasing interest in using measures that reduce the myocardial workload. This article highlights nonpharmacologic approaches to the management of severe heart failure in the critically ill child. It also concentrates on physiologic approaches that address the balance between oxygen demand and delivery; the manipulation of cardiopulmonary interactions to optimize ventricular function; and the use of mechanical circulatory support as a method of achieving ultimate myocardial rest. PMID- 11389359 TI - Pediatric sepsis and multiple organ dysfunction syndrome. AB - Systemic inflammatory response syndrome may be viewed as the systemic expression of cytokine signals that normally function on an autocrine or paracrine level. Sepsis is defined as systemic inflammatory response syndrome caused by an infection. Multiple organ dysfunction syndrome may represent the end stage of severe systemic inflammatory response syndrome or sepsis. Many cells are involved, including endothelial cells and leukocytes and multiple proinflammatory and antiinflammatory mediators (cytokines, oxygen free radicals, coagulation factors, and so forth). Various pathophysiologic mechanisms have been postulated. The most popular theory is that the inflammatory process loses its autoregulatory capacity; however, microcirculatory dysregulation and apoptosis may also be important, and a new paradigm posits a complex nonlinear system. Many new treatments have been studied recently. The usefulness of immune modulating diets remains to be evaluated. Molecular immunomodulation is still of unclear value. The therapy of sepsis and multiple organ dysfunction syndrome remains mainly supportive. PMID- 11389361 TI - Bronchiolitis in infants. AB - Bronchiolitis is a common cause of wheezing among infants. Respiratory syncytial virus (RSV) is the most common infectious agent to cause bronchiolitis, and RSV infection accounts for more than 125,000 hospitalizations per year in the United States. Beyond supportive measures, the care of infants with bronchiolitis remains controversial. Practitioners continue to treat infants with a variety of pharmacologic agents, despite limited evidence of their efficacy. Investigators continue to search for the safest and most cost-effective methods to treat infants with bronchiolitis, not only to overcome obstructive symptoms during the acute illness, but also to prevent recurrent symptoms of airway obstruction that occur in some children for years after their initial episode of bronchiolitis. Improved understanding of the pathogenesis of RSV infection and of virus-host interactions may one day lead to the development of agents that alter the initial inflammatory response and strategies that help prevent recurrent episodes of wheezing and the development of asthma after acute bronchiolitis. PMID- 11389362 TI - Current concepts in adult respiratory distress syndrome in children. AB - Acute respiratory distress syndrome (ARDS) is an acute form of severe alveolar capillary injury that evolves after a direct or indirect lung insult. It begins as noncardiogenic pulmonary edema and develops into a neutrophilic alveolitis, and, later, pulmonary fibrosis. Mortality remains high among children with ARDS, particularly when serious underlying conditions co-exist, sepsis occurs, and when there is multi-organ failure. Lung function improves with time among survivors, but pulmonary fibrosis may persist. Advances in the care of children with ARDS include the use of lung-protective ventilator strategies, permissive hypercapnia, inhaled nitric oxide, high-frequency ventilation, and extra-corporeal life support. These approaches reduce ventilator-associated lung injury and may improve survival when used in combination with one another. Interventions that reduce alveolar inflammation, enhance alveolar fluid removal, and reduce pulmonary fibrosis will further improve survival and recovery from ARDS in the future. PMID- 11389363 TI - Current concepts on the genetics of asthma. AB - Asthma is a complex genetic disorder with variable phenotype, largely attributed to the interactions of the environment and multiple genes, each potentially having small effects. Numerous asthma and atopy loci have been reported in studies demonstrating associations and/or linkage of the asthma-associated phenotypes, atopy, elevated IgE levels, and bronchial hyperresponsiveness to alleles of microsatellite markers and single nucleotide polymorphisms within specific cytokine/chemokine and IgE regulating genes. Although the studies reporting these observations are compelling, most of them lack statistical power. This review compiles the evidence that supports linkage and associations to the various genetic loci and candidate genes. Whereas significant progress has been made in the field of asthma genetics in the past decade, the roles of the genes and genetic variations within the numerous candidate asthma genes that have been found to associate with the expression of the asthmatic phenotype remain to be determined. PMID- 11389365 TI - Updates in pediatric nutrition. AB - Ongoing research in several areas of pediatric nutrition has new practical applications for community-based pediatricians. For example, a fresh understanding of risk factors for rickets persuades pediatricians to recognize and treat this disease, which was thought to be nearly extinct in the modern industrialized world. Similarly, an expanded awareness of the antibacterial components of breast milk encourages a more complete dialogue between pediatricians and new mothers about the potential benefits of breast-feeding. For those infants with feeding intolerance, new data help to refine the indications for hypoallergenic formulas, which are increasingly recommended for children with a variety of symptoms. The past year also has seen breakthroughs in our understanding of supplemental nutrition for children. Vitamin A may provide direct benefits for the most vulnerable of children, namely premature infants at high risk for lung disease. At the other end of the pediatric spectrum, adolescent athletes seeking to enhance their performance are consuming poorly studied sports supplements that may not be beneficial and may even be toxic. Finally, a greater appreciation for the epidemic of obesity that is sweeping the United States and other countries suggests that children at high risk may represent a far more diverse population than had been recognized previously. PMID- 11389366 TI - Dental health for the pediatrician. AB - Pediatricians play an important role in promoting oral health care, and their advice regarding dental procedures or therapies may be solicited. This review is intended to provide answers for the more common questions that parents may have regarding the oral health of their children. The controversies surrounding early orthodontic therapy and mercury-containing amalgam fillings continue to be areas of contention. Why does the orthodontist want to initiate orthodontic treatment in a 7-year-old child? Is there justification for the banning of amalgam fillings, as has occurred in some countries? Also explored are the current therapies for two of the more common oral mucosal disorders: aphthous ulcers and recurrent herpes simplex labialis. These subjects have relevance not only for pediatric patients but also for adults (and for many providers) as well. PMID- 11389367 TI - Biomechanical evaluation of occipitocervical fixation devices. AB - Human cadaveric occipitocervical specimens were implanted with three types of instrumentation. The devices were tested biomechanically under three modes of loading to determine the stiffness of spinal constructs and the failure mechanisms of the constructs under extreme flexion. The devices tested were the AXIS Fixation System (with custom plate), the Y-Plate, and the Luque rectangle. No significant differences in stiffness among the devices were found under compression and flexion. The stiffnesses of the plate systems were statistically higher than the Luque rectangle in extension and torsion. In extreme flexion, the plate systems failed by fracture of the C2 pedicles. Modern plate systems, for occipitocervical fixation, provide more stiffness and stability than traditional wiring techniques. This study provides surgeons with information on the relative merits of modern plate and screw systems compared with traditional rod and wire constructs. PMID- 11389368 TI - Distraction extension injuries of the cervical spine. AB - Twenty-four consecutive patients with cervical distraction extension injuries were retrospectively reviewed to study the safety and efficacy of various treatment protocols in this type of cervical spine injury. Sixteen of 24 patients with cervical distraction extension injuries underwent surgical stabilization. All patients undergoing surgical stabilization were noted to have a stable fusion at their latest follow-up. There were three instances of surgically related neurologic deterioration as a result of over-distraction of the anterior column interspace at the time of graft placement. The overall mortality rate was 42% in this aged patient population. Anterior reconstruction of the cervical spine with an anterior cervical graft and plate acting as a tension band is the ideal treatment method for stabilization of acute distraction extension injuries involving primarily the soft tissue structures (anterior longitudinal ligament and intervertebral disc). Type 2 injuries, depending on the degree of displacement and the adequacy of closed reduction, may need to be approached initially posteriorly to obtain adequate alignment, followed by an anterior reconstructive procedure. Great care should be taken during anterior graft placement to avoid over-distraction of the spine. If nonsurgical intervention is selected, close regular radiographic follow-up is necessary to detect early vertebral malalignment, which may predispose to spinal cord dysfunction. Older patients sustaining this injury have a high mortality rate. PMID- 11389369 TI - Nonlinear finite-element analysis of the lower cervical spine (C4-C6) under axial loading. AB - This study was conducted to develop a detailed, nonlinear three-dimensional geometrically and mechanically accurate finite-element model of the human lower cervical spine using a high-definition digitizer. This direct digitizing process also offers an additional method in the development of the finite-element model for the human cervical spine. The biomechanical response of the finite-element model was validated and corresponded closely with the published experimental data and existing finite-element models under axial compressive loading. Furthermore, the results indicated that the cervical spine segment response is nonlinear with increasing stiffness at higher loads. As a logical step, a parametric study was conducted by evaluating the biomechanical response related to the changes in the modeling techniques of the finite-element model and the mechanical properties of the disk annulus. Variations of the predicted horizontal disk bulge were investigated under axial compressive displacements for the normal model, the model without facet articulations, and the model without nucleus. Removal of nucleus fluids causes an inward bulge of the inner annulus layers, with the displacement magnitude dependent on external loads. The result indicates that the nucleus fluid plays an important role in cervical spine mechanics. Simulated facetectomy indicates a decrease in the stiffness of the cervical spine. The study shows that, in reality, the stiffness of the lower cervical spine depends closely on factors such as the spinal geometry and physical properties, thereby resulting in various force and displacement responses. PMID- 11389370 TI - Penetration of cranial inner table with Gardner-Wells tongs. AB - The compression force exerted by Gardner-Wells tongs was compared with the force necessary to penetrate the cranial inner table with a Gardner-Wells tong pin. Load cells measured the force exerted by the spring-loaded Gardner-Wells tong pin on fresh cadaveric skull samples. Increasing forces were exerted until penetration of the inner table occurred. At the manufacturer's recommended, 1 mm indicator stem protrusion an average of 30 pounds of compressive force was exerted by the spring-loaded pin. The average force necessary to penetrate the inner table with the cadaveric specimens with the tong pin was 162 pounds. The force necessary to penetrate the inner table of the skull well exceeds that exerted by properly applied tongs, suggesting that the risk of inner table penetration is low. PMID- 11389371 TI - Lumbar posterolateral fusion with biphasic calcium phosphate ceramic. AB - The authors conducted a retrospective observational study of patients who needed lumbar posterolateral fusion (PLF) using a biphasic calcium phosphate ceramic implant as a substitute for bone graft. The findings of clinical, radiographic, and histologic examinations were reviewed. Thirty-two patients underwent single level PLF with instrumentation. In all cases, to decrease the occurrence of donor site complications and morbidity, locally harvested morselized bone from the decompressive site was mixed with hydroxyapatite and beta-tricalcium phosphate (HAP-TCP) granules and sticks and used for fusion at the posterolateral aspect of the lumbar spine. The histologic findings of three biopsy specimens obtained during second operations for metallic implant removal were reviewed. The minimum follow-up period was 26 months. There was no evidence of instrument loosening or breakage. However, bone-graft incorporation was difficult to evaluate radiographically, because image quality was inferior to that with conventional autogenous iliac bone graft. Partial graft bone resorption was revealed on radiographs in 75% of cases. The results showed clinical improvement in all but one case. Solid fusion was observed during the second operation in all three cases. Histologic analysis showed excellent bone incorporation around the HAP-TCP granules. These findings suggest that, although the bulk of the fusion mass with HAP-TCP was smaller than that with autogenous bone, this technique combined with rigid instrumentation is a safe and effective procedure. PMID- 11389372 TI - A comparison of fusion rates between single-level cervical corpectomy and two level discectomy and fusion. AB - A single corpectomy and strut grafting has been proposed as an alternative to performing two-level adjacent discectomies with multiple grafts to produce superior fusion rates. The purpose of this study was to compare the clinical and radiographic success of two-level discectomy and fusion with anterior cervical plate fixation compared with a single-level corpectomy. Fifty-two patients were treated with either a two-level adjacent anterior cervical discectomy and fusion with cervical plating, or by a single-level corpectomy and plate. Thirty-two patients had two-level discectomies, whereas 20 had a single corpectomy and a strut graft (average follow-up was 3.6 years). One patient had a pseudarthrosis from a single-level corpectomy and required subsequent surgery to obtain an osseous union. The fusion rates between the two groups was not statistically significant (p = 0.385). The clinical results of the surgeries were similar between the groups based on Odom's criteria. The addition of cervical plates to either two-level discectomies or single-level corpectomies yielded similar fusion and complication rates. PMID- 11389373 TI - The sagittal profile of the cervical and lumbosacral spine in Scheuermann thoracic kyphosis. AB - The sagittal profiles of the cervical and lumbar spine have not been studied in Scheuermann kyphosis. The purpose of this study was to investigate these profiles. Standing lateral radiographs of the spine in 34 children with Scheuermann kyphosis were reviewed. Cervical lordosis, lumbar lordosis, thoracic kyphosis, sagittal vertebral axis, and sacral inclination were measured. The relations between these variables were explored using the Pearson correlation. The average patient age was 15.5 +/- 1.8 years, thoracic kyphosis was 65 degrees +/- 12 degrees, lumbar lordosis 71 degrees +/- 13 degrees, and cervical lordosis 4 degrees +/- 15 degrees (Cobb angle), and 9 degrees +/- 14 degrees (posterior vertebral body angle [PVBA]). No correlations were noted between cervical lordosis and thoracic kyphosis. Correlations were noted between cervical lordosis and lumbar lordosis (r2 = 0.17, Cobb angle; r2 = 0.16, PVBA) and between cervical lordosis and the residual sagittal difference (thoracic kyphosis minus lumbar lordosis; r2 = 0.32, p = 0.001 [Cobb angle], and r2 = 0.19, p = 0.01 [PVBA]). In Scheuermann kyphosis, the flexible cervical and lumbar spine is linked by the intermediate rigid thoracic segment. As the residual sagittal difference becomes more kyphotic, lordosis of the cervical spine increases as the patient strives to maintain a forward visual gaze. PMID- 11389374 TI - Mechanical testing of a single rod versus a double rod in a long-segment animal model. AB - This study involved the mechanical testing of single-rod segmental hook fixation and double-rod segmental hook fixation in a long-segment animal model. The goals were first to compare the flexibility of a single-rod scoliosis construct with that of a double-rod construct when tested in torsion, and second, to determine the effect of not using instrumentation with every vertebral segment for the single rod. Another study found that the single-rod construct was as stiff in torsion as the standard double-rod construct in a model of 10 vertebral segments. The amount of neutral zone (NZ) rotation was tested in five calf spines using an MTS (Material Testing System) machine. Five constructs were tested and included 1) a single rod with hooks at every level except the apex; 2) a single rod with two fewer hooks; 3) a single rod with four fewer hooks; 4) a double-rod construct; and 5) no instrumentation. The amount of NZ rotation between vertebral segments was measured over 12, 10, 8, 6, 4, and 2 vertebral segments. An analysis of variance with all constructs showed that the instrumented spines had significantly less movement than did the uninstrumented spine. Statistical comparison using analysis of variance of constructs (constructs 1 to 4) showed that over 12 vertebral segments (T4-L3), all single-rod constructs (constructs 1 to 3) allowed more NZ rotation than did the standard double-rod construct. This testing indicated that over 12 vertebral segments the single rod allowed more NZ rotation than a double-rod construct. PMID- 11389375 TI - Total spondylectomy for primary malignant, aggressive benign, and solitary metastatic bone tumors of the thoracolumbar spine. AB - The records of 14 patients with malignant or aggressive benign vertebral tumors of the thoracolumbar spine who underwent total spondylectomy (TS) were evaluated retrospectively. Total spondylectomy was performed by bisecting the affected vertebra through the pedicle using fine threadwire saws and removing the vertebra en bloc through the posterior procedure alone or the one-stage anteroposterior combined procedure. Remarkable pain relief and ambulation after surgery were achieved in all 14 patients. No serious complications occurred. Nerve roots were sacrificed in seven cases. A marginal surgical margin was achieved in 10 cases and an intralesional surgical margin was achieved in four. At the site of the osteotomized pedicle, the surgical margin was marginal, with the possibility of tumor-cell contamination in 10 cases. Local recurrence was found in three cases of posterior total spondylectomy at 0.3 to 3.5 years (mean, 3.2 years) follow-up evaluation at the other site of the osteotomized pedicle. These results suggest that this type of total spondylectomy is effective in controlling local recurrence without incurring major complications and is a clinically useful procedure. PMID- 11389376 TI - Effect of stimulus pulse duration on intraoperative somatosensory evoked potential (SEP) monitoring. AB - The effect of the stimulus duration on the amplitude and latency of intraoperative somatosensory evoked potential (SEP) was studied in 30 patients with idiopathic scoliosis undergoing surgery. Constant current square-wave electrical stimulation was applied to the posterior tibial nerve at a rate of 5.1 Hz. The effects of both the pulse duration and the stimulus current density on the intraoperative SEP were evaluated. Amplitudes and latencies of SEP were analyzed by one-way parametric analysis of variance. SEP signal recording was found to be difficult if the stimulus duration was less than 0.05 ms. The stimulus duration had no significant effect on the latencies of the SEP, but the amplitude of the SEP showed significant changes with differing stimulus durations. The SEP amplitudes were found with significant increases in pulse durations less than 0.3 ms. Stimulus pulse duration has a significant effect on SEP amplitude, and this should be taken into consideration during intraoperative SEP monitoring. A pulse duration of 0.3 ms is recommended for SEP using posterior tibial nerve stimulation. PMID- 11389377 TI - Modeling of the naked facet sign in the thoracolumbar spine. AB - On transverse computed tomographic (CT) scan cuts of the thoracolumbar spine, the naked facet sign occurs when the inferior articular facets of the cephalad vertebra do not appear adjacent to the superior facets of the subjacent caudal vertebra. The objective of this study was to determine the angles of rotation required for the naked facet sign to occur in the thoracolumbar spine, with the center of rotation located at various points in or anterior to the vertebral body. A commercial spinal model and visualization software were used to simulate various flexion injuries. Each functional spinal unit (FSU; T11-T12, T12-L1, and L1-L2) was examined separately. In the model, two CT scan slices (each 2 mm thick) were created parallel to the inferior end plate of the cephalad vertebra of each FSU. The cephalad vertebra was rotated in 0.5 degrees increments, and after rotation both modeled CT slices were examined for the presence of the naked facet sign. If the sign did not occur, the process was repeated, rotating the cephalad vertebra an additional 0.5 degrees until the naked facet sign occurred. The angle of rotation necessary for the sign to occur increases as the point of rotation of the vertebra moves from anterior to posterior and from superior to inferior. The naked facet sign occurred at a minimum rotation angle of 5 degrees (with respect to the anterior-superior point on T11) and at a maximum rotation angle of 16.5 degrees (with respect to the posterior-inferior point on L1). For rotations about a point located 3 cm anterior to the vertebral body, the minimum angles required for the sign decreased only 1 degrees for each FSU. These results suggest that the naked facet sign does not consistently imply the presence of posterior column vertebral instability. This will help clinicians to relate the mechanism of injury, radiographic findings (including the naked facet sign), and the implied injury pattern to the determination of stability, and ultimately the management options for the injury. PMID- 11389378 TI - Horizontal intervertebral cage fusions. PMID- 11389379 TI - Retroperitoneal malignant peripheral nerve sheath tumor associated with scoliosis in neurofibromatosis. AB - Neurofibromatosis 1 is an autosomal dominant disorder characterized by distinctive clinical problems including scoliosis and malignant peripheral nerve sheath tumors. We present two cases of retroperitoneal malignant peripheral nerve sheath tumor associated with scoliosis in neurofibromatosis. Presence of spinal deformity resulted in delay of the diagnosis of the sarcoma. PMID- 11389380 TI - Stab injury of the spinal cord surgically treated. AB - The authors report a case of thoracic spinal cord stab injury with neurologic impairment that was treated surgically after injury. A literature review and case analysis indicate that surgical extraction of foreign bodies retained within the spinal canal is indicated to avoid infection, delayed myelopathy, and neurologic loss. The amount of motor and functional recovery for incomplete injuries after spinal cord stab wound can be strikingly good despite pathologic changes to severely damaged areas, and removal of retained intraspinal metallic fragment can improve this neurologic outcome. Open removal of the knife seems preferable to avoid bleeding and infection. PMID- 11389381 TI - Pancreatitis after posterior spinal fusion for adolescent idiopathic scoliosis. AB - A case report describes an adolescent with idiopathic scoliosis who underwent a posterior spinal fusion and developed pancreatitis postoperatively. The patient recovered with parenteral nutrition support. We report this case to add to the literature that supports a benign disease course for postoperative pancreatitis in patients who have had posterior spinal fusion. PMID- 11389382 TI - Development and implementation of a clinical pathway for spinal cord injuries. AB - The authors have developed a clinical pathway for the treatment of spinal cord injuries to help improve patient care. A clinical pathway for the treatment of patients with spinal cord injury was developed through a multidisciplinary approach. The control group (group 1) consisted of patients who were treated in the 2 years before the initiation of the pathway. Data from patients treated in conjunction with this pathway were collected prospectively (group 2). Thirty-six patients were treated in conjunction with the pathway compared with 22 in the control group. Group 2 had 6.8 fewer intensive care unit days, 11.5 fewer hospital days, 6 fewer ventilator days (p < 0.05), and a lower rate of complications. The use of a clinical care pathway for spinal cord injuries has resulted in improved patient care and fewer complications. PMID- 11389384 TI - The vascular anatomy anterior to the L5-S1 disk space. AB - STUDY DESIGN: Dissection of 37 human cadavers was performed to assess the variability in the vascular anatomy anterior to the L5-S1 disc space. OBJECTIVES: To determine the variability of the anterior vascular anatomy at the L5-S1 disc space, and to assess its reliability as an anatomic landmark for the placement of anterior interbody fusion devices. SUMMARY OF BACKGROUND DATA: Although multiple studies have defined both the lumbar spinal anatomy and the anatomy of the great vessels, the relation of the great vessels to the anterior L5-S1 disc space has not been quantified directly. METHODS: This study investigated 35 human cadavers (17 males and 18 females). The anterior L5-S1 disc space and great vessel bifurcation were exposed through a transabdominal approach. Two independent observers each obtained 10 measurements in each specimen. RESULTS: The middle sacral artery was present in 100% of the specimens, averaging 2.5 mm in width. Its location in relation to the midline was quite variable, with a range greater than 2 cm in both the top and bottom of the disc. The distance from the bifurcation to the top of the L5-S1 disc averaged 18 mm (range, 7-36 mm). The total width between the left common iliac vein and the right common iliac artery averaged 33.5 mm (range, 12-50 mm). CONCLUSIONS: The middle sacral artery, present in 100% of the specimens, is a poor anatomic landmark for locating the midline at L5-S1. Because the average space available between the left common iliac vein and the right common iliac artery is 33.5 mm, and because the left common iliac vein averages only 12 mm from midline, the surgeon must be prepared to mobilize the local vasculature in most cases to expose the L5-S1 disc space adequately. PMID- 11389385 TI - Immunophenotypic analysis of the inflammatory infiltrates in herniated intervertebral discs. AB - STUDY DESIGN: The herniated portion of the lumbar disc was analyzed immunohistochemically for inflammatory infiltrates to determine their immunophenotype. OBJECTIVE: To investigate the pathomechanism behind spontaneous regression of herniated discs. SUMMARY OF BACKGROUND DATA: Spontaneous regression of herniated intervertebral discs has been increasingly reported. The inflammatory response of the host has been suggested as a factor in this phenomenon. However, whether the inflammation is induced from direct chemical irritation of the nucleus pulposus material or whether it is secondary to an autoimmune response to the nucleus pulposus remains controversial. METHODS: The herniated portion of the disc was collected from 38 patients who underwent surgery for lumbar disc herniation. Thin cryostat sections were made, and the extent to which inflammatory cells had infiltrated the disc specimen was defined. Then the immunophenotype of cellular infiltrates in the herniated disc specimens was assessed by immunostaining using a series of antibodies for lymphocyte, monocyte, macrophage, and dendritic cell markers. RESULTS: The inflammatory infiltrates in 14 of the 38 herniated discs were subjected to immunohistochemical analysis. None of them expressed the immunophenotypic markers of the lymphocyte (CD20, CD45RO, CD4, CD8, TCRgammadelta), mature monocyte (CD33), or dendritic cell (CD1a, CD80, CD86, S100). Abundant infiltration of CD68-positive cells that lacked CD33 but had a variable amount of CD11b, CD11c, and CD40 likely represents a process of differentiation from monocytes to macrophages. CONCLUSIONS: These findings are consistent with an immunophenotype of inflammatory responses to tissue injury or chemical irritation rather than antigen-specific immune responses. Therefore, understanding the mechanism of tissue repair is fundamentally important in the management of patients with disc herniations. PMID- 11389387 TI - Biomechanical factors influencing nuclear disruption of the intervertebral disc. AB - STUDY DESIGN: A disc model with full anular division was used to investigate how different biomechanical parameters influence the severity of nuclear disruption during compressive loading. OBJECTIVE: To quantify the manner in which flexion, hydration, and loading rate contribute to the breakdown in the intrinsic cohesive structure of the nucleus pulposus. SUMMARY OF BACKGROUND DATA: The risk of disc herniation is known to increase when the disc is loaded in flexed positions. However, there is a lack of experimental data showing how a combination of flexion with different loading rates and hydration levels affects the extent of nuclear disruption. METHODS: A reproducible state of full hydration was established for isolated bovine caudal discs. A period of static preloading at an applied stress of 1 MPa was used to obtain a consistent state of partial hydration. Then 96 discs were subjected to a full-thickness division of the anulus fibrosus and compressed while hydration level, degree of flexion, and rate of loading were varied systematically. RESULTS: A full spectrum of nuclear damage was observed in the tests, ranging from no detectable disruption to sudden sequestration of the entire nucleus. These results were quantified, and a general correlation was established between the severity of disruption and the different loading parameters. CONCLUSIONS: The degree of flexion and the level of hydration were shown to play an important role in influencing the tendency of the nucleus to break loose and extrude through a preexisting anular division. Interestingly, the rate of loading appeared to have only a minor effect on the severity of damage induced in discs that incorporated a full depth anular division. PMID- 11389386 TI - Functional recovery and regeneration of descending tracts in rats after spinal cord transection in infancy. AB - STUDY DESIGN: The functional recovery of rats that underwent spinal cord transection in infancy was evaluated by multimodal examination (functional tests, electrophysiologic evaluation, tract-tracing) to determine the basis for the recovery. OBJECTIVES: To determine whether the hind limb function in rats that underwent spinal cord transection in infancy is regained completely, which descending tracts regenerate after the transection, and whether the functional recovery is correlated with axonal reconnection. SUMMARY OF BACKGROUND DATA: It is widely accepted that a newborn rat recovers its hind limb function after spinal cord transection even without any specific treatments. This functional recovery might be attributed to possible regeneration of some descending pathways, although there is a counterargument that well-trained spinal cord reflexes may bring about functional compensation. METHODS: The thoracic spinal cord of infant rats was completely transected at Th10 when they were 2 weeks of age. Multimodal functional tests and electrophysiologic studies were performed 5 weeks later. Some recovered rats (i.e., those able to walk after the transection) underwent spinal cord retransection, with subsequent reevaluation of locomotion and muscle-evoked potentials. At 6 weeks after the initial transection, tract tracing studies were performed in some animals. RESULTS: A motor performance score detected the functional differences between the control and the recovered rats. Muscle-evoked potentials of hind limbs after electrical stimulation to the brain were recorded in some of the recovered rats, but never in the unrecovered rats. Moreover, the muscle-evoked potentials of the recovered rats disappeared after spinal cord retransection that resulted in loss of voluntary movement. Morphologic studies in two rats provided evidence that reconnection of rubrospinal, vestibulospinal, and reticulospinal tracts had occurred, whereas corticospinal regeneration was not detected. CONCLUSIONS: It can be concluded that the hind limb function of rats that underwent spinal cord transection in infancy was partially regained; that axonal regeneration of the rubrospinal, vestibulospinal, or reticulospinal tracts was demonstrated, whereas the reconnection of the corticospinal tract was not observed; and that the axonal regeneration of these tracts is involved in the functional recovery. PMID- 11389388 TI - Canal and intervertebral foramen encroachments of a burst fracture: effects from the center of rotation. AB - STUDY DESIGN: The neural spaces of thoracolumbar burst fractures were investigated in an in vitro biomechanical study. OBJECTIVE: To evaluate encroachments of spinal canal diameter and intervertebral foramen area as functions of where the center of rotation is located during flexion and extension. SUMMARY OF BACKGROUND DATA: Decompression of the neural spaces is important for the recovery of neural function in a patient with a burst fracture injury. A few biomechanical studies have documented the decompression of the neural elements by adjustment of posterior fixation devices. However, the device adjustments have been device specific and ill defined. No study has investigated the neural decompression phenomenon with precisely defined multiple adjustments. METHODS: Burst fractures were produced at L1 vertebra in nine T11-L3 human spinal segments. Specimens were flexed and extended around five different centers of rotation located in the mid-L1 plane. The spinal canal diameter and intervertebral foramen area encroachments were quantified in maximum flexion and extension around each center of rotation using lateral radiographs. RESULTS: The average canal encroachment of 42.6% changed in flexion (32.2-48.5%) and extension (36.3-44.2%) by location of the center of rotation. The average intervertebral foramen area encroachment was decreased to a greater extent more often in flexion than in extension because of where the center of rotation was located. CONCLUSIONS: Both flexion and extension can decompress canal and foramina, depending on the choice for the location of the center of rotation. If lordotic posture is preferred clinically, then the optimal choice may be extension around the center of rotation located at the tip of the spinous process of the burst vertebra. PMID- 11389390 TI - Correlation between operative outcomes of cervical compression myelopathy and mri of the spinal cord. AB - STUDY DESIGN: Magnetic resonance images of cervical compression myelopathy were retrospectively analyzed in comparison with surgical outcomes. OBJECTIVES: To investigate which magnetic resonance findings in patients with cervical compression myelopathy reflect the clinical symptoms and prognosis, and to determine the radiographic and clinical factors that correlate with the prognosis. SUMMARY OF BACKGROUND DATA: Signal intensity changes of the spinal cord on magnetic resonance imaging in chronic cervical myelopathy are thought to be indicative of the prognosis. However, the prognostic significance of signal intensity change remains controversial. METHODS: The participants in this study were 73 patients who underwent cervical expansive laminoplasty for cervical compression myelopathy. Their mean age was 64 years, and the mean postoperative follow-up period was 3.4 years. The pathologic conditions were cervical spondylotic myelopathy in 42 patients and ossification of the posterior longitudinal ligament in 31 patients. Magnetic resonance imaging (spin-echo sequence) was performed in all the patients. The transverse area of the spinal cord at the site of maximal compression was computed, and spinal cord signal intensity changes were evaluated before and after surgery. Three patterns of spinal cord signal intensity changes on T1-weighted sequences/T2-weighted sequences were detected as follows: normal/normal, normal/high-signal intensity changes, and low-signal/high-signal intensity changes. Surgical outcomes were compared among these three groups. The most useful combination of parameters for predicting prognosis was determined using a stepwise regression analysis. RESULTS: The findings showed 2 patients with normal/normal, 67 patients with normal/high-signal, and 4 patients with low-signal/high-signal change patterns before surgery. Regarding postoperative recovery, the preoperative low signal/high-signal group was significantly inferior to the preoperative normal/high-signal group. There was no significant difference between the transverse area of the spinal cord at the site of maximal compression in the normal/high-signal group and the low-signal/high-signal group. A stepwise regression analysis showed that the best combination of surgical outcome predictors included age (correlation coefficient R = -0.348), preoperative signal pattern, and duration of symptoms (correlation coefficient R = -0.231). CONCLUSIONS: The low-signal intensity changes on T1-weighted sequences indicated a poor prognosis. The authors speculate that high-signal intensity changes on T2 weighted images include a broad spectrum of compressive myelomalacic pathologies and reflect a broad spectrum of spinal cord recuperative potentials. Predictors of surgical outcomes are preoperative signal intensity change pattern of the spinal cord on radiologic evaluations, age at the time of surgery, and chronicity of the disease. PMID- 11389391 TI - Headache, neck pain, and neck mobility after acute whiplash injury: a prospective study. AB - STUDY DESIGN: A 6-month prospective study of neck mobility in patients with acute whiplash injury and a control group with acute ankle distortion was conducted. OBJECTIVES: To assess active neck mobility after acute whiplash and ankle distortion injuries, and to relate neck mobility to headache, neck pain, and speed of car at the time of collision. SUMMARY OF BACKGROUND DATA: A major problem after whiplash injury is restriction of neck mobility immediately subsequent to trauma. It is, however, unclear whether neck mobility changes after the acute injury are related to the associated headache and neck pain. METHODS: Cervical range of neck motion, neck pain, and headache were assessed after 1 week, then 1, 3, and 6 months after injury in 141 patients with acute whiplash injury, and in 40 patients with acute nonsport ankle distortion. RESULTS: Patients with whiplash injury had significantly reduced flexion, extension, lateral flexion, and rotation of the neck immediately after injury, as compared with patients with ankle distortion injury. Neck mobility, however, was similar in the two groups after 3 months. In patients with whiplash injury, neck pain and neck mobility were found to be related inversely to reported headache and neck mobility. Neck mobility was not significantly related to a difference in car speed at the time of collision. CONCLUSIONS: Neck mobility is reduced immediately after, but not 3 months after, a whiplash trauma. Headache and neck mobility are related inversely and neck pain and neck mobility are related inversely during the first 6 months after acute whiplash injury. PMID- 11389392 TI - Whiplash syndrome: kinematic factors influencing pain patterns. AB - STUDY DESIGN: The overall, local, and segmental kinematic responses of intact human cadaver head-neck complexes undergoing an inertia-type rear-end impact were quantified. High-speed, high-resolution digital video data of individual facet joint motions during the event were statistically evaluated. OBJECTIVES: To deduce the potential for various vertebral column components to be exposed to adverse strains that could result in their participation as pain generators, and to evaluate the abnormal motions that occur during this traumatic event. SUMMARY OF BACKGROUND DATA: The vertebral column is known to incur a nonphysiologic curvature during the application of an inertial-type rear-end impact. No previous studies, however, have quantified the local component motions (facet joint compression and sliding) that occur as a result of rear-impact loading. METHODS: Intact human cadaver head-neck complexes underwent inertia-type rear-end impact with predominant moments in the sagittal plane. High-resolution digital video was used to track the motions of individual facet joints during the event. Localized angular motion changes at each vertebral segment were analyzed to quantify the abnormal curvature changes. Facet joint motions were analyzed statistically to obtain differences between anterior and posterior strains. RESULTS: The spine initially assumed an S-curve, with the upper spinal levels in flexion and the lower spinal levels in extension. The upper C-spine flexion occurred early in the event (approximately 60 ms) during the time the head maintained its static inertia. The lower cervical spine facet joints demonstrated statistically greater compressive motions in the dorsal aspect than in the ventral aspect, whereas the sliding anteroposterior motions were the same. CONCLUSIONS: The nonphysiologic kinematic responses during a whiplash impact may induce stresses in certain upper cervical neural structures or lower facet joints, resulting in possible compromise sufficient to elicit either neuropathic or nociceptive pain. These dynamic alterations of the upper level (occiput to C2) could impart potentially adverse forces to related neural structures, with subsequent development of a neuropathic pain process. The pinching of the lower facet joints may lead to potential for local tissue injury and nociceptive pain. PMID- 11389393 TI - The arthrodesis rate in multilevel anterior cervical fusions using autogenous fibula. AB - STUDY DESIGN: This study was a roentgenographic review of 145 patients who underwent multilevel anterior cervical arthrodesis using autogenous fibula. OBJECTIVE: To determine the arthrodesis rate in multilevel cervical fusions using autogenous fibula. SUMMARY OF BACKGROUND DATA: Previous studies have shown an unacceptably high rate of nonunions with multilevel anterior cervical arthrodesis. However, this has not been the clinical experience of the author's group. METHODS: Lateral roentgenograms taken a minimum of 2 years after surgery were reviewed independently by two radiologists. If either radiologist questioned any fusion level, the final decision was made on the basis of flexion-extension roentgenograms. RESULTS: An overall union rate of 90% and 94% per patient was found for each level of attempted fusion. A solid arthrodesis was achieved in 93% of the patients with two-level fusions, and 84% of the three-level fusions were solid. The difference was not statistically significant. CONCLUSION: Autogenous fibula used as a strut graft results in an acceptable union rate for multilevel anterior cervical arthrodesis. PMID- 11389394 TI - Association between a polymorphism of the transforming growth factor-beta1 gene and genetic susceptibility to ossification of the posterior longitudinal ligament in Japanese patients. AB - STUDY DESIGN: A study was conducted to determine the association between polymorphism of the transforming growth factor-beta1 (TGF-beta1) gene and ossification of the posterior longitudinal ligament (OPLL) prevalence. OBJECTIVE: To examine whether the T869-->C polymorphism of the TGF-beta1 gene is associated with genetic susceptibility to OPLL in Japanese subjects. SUMMARY OF BACKGROUND DATA: In the posterior longitudinal ligament, OPLL is associated with abnormal calcium metabolism. Several candidate genes are associated with the prevalence of OPLL. In the ossified matrix and chondrocytes of adjacent cartilaginous areas of OPLL, TGF-beta1 is overexpressed. METHODS: The TGF-beta1 genotype was identified with an allele-specific polymerase chain reaction method in 319 Japanese subjects (46 subjects with OPLL and 273 control subjects). RESULTS: There was a significant association between the T869-->C genotype and the prevalence of OPLL in the cervical spine. Multivariable logistic regression analysis, adjusted for gender, age, height, and body weight, showed that the frequency of the C allele was significantly higher in subjects with OPLL than in control subjects. CONCLUSIONS: The T869-->C polymorphism of the TGF-beta1 gene is a genetic determinant of a predisposition to OPLL, with the C allele representing a risk factor for genetic susceptibility to OPLL in Japanese subjects. Therefore, TGF beta1 genotyping may be useful in the prevention of OPLL. PMID- 11389396 TI - Patient outcomes after Harrington instrumentation for idiopathic scoliosis: a 15- to 28-year evaluation. AB - STUDY DESIGN: A retrospective study was performed, using the Short Form-36 Health Survey and the Roland and Morris Disability Questionnaire, to investigate patient outcomes after fusion for adolescent idiopathic scoliosis using Harrington rod instrumentation. OBJECTIVE: To evaluate health-related quality of life and low back pain in a long-term follow-up study of surgery for adolescent idiopathic scoliosis. SUMMARY OF BACKGROUND DATA: The commonly accepted surgical treatment for idiopathic evolutive scoliosis is vertebral fusion. It has been suggested that this procedure may cause low back pain and a poor quality of life over the long term. Outcome measures after surgery for adolescent idiopathic scoliosis have focused mainly on objective parameters such as radiographic measures. However, this information has proved to be correlated only weakly with outcomes that are more relevant to patients, such as functional status and symptoms. Until recently, only a few long-term outcome studies have used standardized and validated patient-oriented tools to evaluate surgically treated patients with scoliosis. METHODS: In this study, 70 patients treated with a standard Harrington technique were recontacted and evaluated by means of self-administered questionnaires (Short Form-36 Health Survey and Roland and Morris Disability, clinical examination, and radiographic analysis. Preoperative and follow-up radiographic findings were registered. Relations between radiographic and patient oriented data were evaluated. RESULTS: A comparison between the current sample and the Italian age-matched normative data for the Short Form-36 Health Survey showed them to have a similar pattern. Findings showed the patient-oriented outcome to be correlated inversely with the extension of vertebral fusion and the preoperative Cobb angle. CONCLUSION: Long-term follow-up evaluation of Harrington rod fusion for adolescent idiopathic scoliosis showed no important impairment of health-related quality of life, as measured by patient-oriented evaluation. PMID- 11389397 TI - Functional classification of patients with idiopathic scoliosis assessed by the Quantec system: a discriminant functional analysis to determine patient curve magnitude. AB - STUDY DESIGN: A stepwise discriminant analysis was used to define a spinal deformity score based on three-dimensional measurements by the Quantec spinal image system (raster stereophotograph). OBJECTIVE: To provide functional classification of spinal deformity in patients with mild idiopathic scoliosis without using radiographs. SUMMARY OF BACKGROUND DATA: Most studies classify the degree of spinal deformity in terms of coronal plane radiograph without analyzing transverse rotation. To the authors' knowledge, no studies investigating classification of spinal deformity in idiopathic scoliosis using Quantec system measurements have been documented. METHODS: In this study, 129 patients with a single curve and 119 patients with a double curve were divided into three groups according to Cobb angle: Group 1 (less than 10 degrees ), Group 2 (10-20 degrees ), and Group 3 (greater than 20 degrees ). RESULTS: The patients were assigned to the group with the highest scores after application of a stepwise discriminant analysis. The accuracy of the classification system by functional scores for the patients with a single curve was 85% for Group 1, 63.5% for Group 2, and 71.7% for Group 3. The accuracy of classification by functional scores for the patients with a double curve was 87.1% for Group 2 and 76.1% for Group 3. CONCLUSION: The back surface image study is a method for providing a quantitative assessment of mild spinal deformity, allowing evaluation of patients by integrated three dimensional parameters with no reference to radiographs. PMID- 11389399 TI - Postural hygiene program to prevent low back pain. AB - STUDY DESIGN: A quasi-experimental study with 3 x 4 design was performed. OBJECTIVE: To improve the level of knowledge and motor skills and thereby avert the development of painful symptoms. SUMMARY OF BACKGROUND DATA: Despite the fact that low back pain affects a high percentage of the population, little research has been carried out to prevent low back pain through health education. METHODS: The participants in this study were 106 third-grade (9-year-old) students. The program included 11 sessions. The teacher attended the placebo group sessions. No intervention was used with the control group. RESULTS: The level of knowledge and motor skills in the experimental group showed a significant increase immediately after the intervention finished, and at 6- and 12-month intervals after completion of the postural hygiene program (P = 0.00). Some positive changes were generalized to natural situations (P = 0.00). In an independent health check carried out by the local school health services 4 years after application of the postural hygiene program, the results tended slightly to favor the experimental condition over the control conditions (placebo + no intervention). A greater number of the control subjects required medical treatment for low back pain, although this difference was only marginally significant (P = 0.07). CONCLUSIONS: The findings from this study support the hypothesis that programs involving practice and motivating strategies impart health knowledge and habits more efficiently than those restricted to the mere transmission of information. PMID- 11389400 TI - Tandem interbody fusion grafting after cervical vertebrectomy. AB - STUDY DESIGN: A case is presented with clinical and radiologic follow-up assessment to evaluate the possible effectiveness of tandem interbody fusion grafting. OBJECTIVE: To design a technique for rescuing a long iliac crest bone autograft that is too short or must be shortened because of the undesirable shape some long iliac crest grafts can take. SUMMARY OF BACKGROUND DATA: Supplementing a larger piece of autograft with a smaller piece in tandem is suggested in this report as a potentially valuable technique for a surgeon presented with a large but inadequate piece of autograft. METHODS: Instead of requiring a second incision to remove iliac crest from the other side or an allograft, the technique described in this report uses a small piece of iliac crest laid in tandem with the original strut graft to span the vertebrectomy channel. RESULTS: A case of an anterior cervical vertebrectomy using a tandem strut graft resulted in good clinical and radiographic results. CONCLUSIONS: Tandem graft placement can salvage a graft that is of inadequate final length. PMID- 11389401 TI - Postoperative pain control using epidural catheters after anterior spinal fusion for adolescent scoliosis. AB - STUDY DESIGN: A prospective review of patients undergoing epidural catheter placement after anterior spinal fusion and instrumentation for adolescent scoliosis was performed. Data were collected using visual analog pain scores reflecting the patients' perception of their pain control. OBJECTIVES: To present the authors' technique for epidural catheter placement and dosing protocol, and to demonstrate the results from postoperative pain control after anterior spinal instrumented fusion for 10 consecutive patients. SUMMARY OF BACKGROUND DATA: The literature regarding the benefits of epidural catheters after spinal surgery is contradictory, even with controlled studies comparing epidural catheters with intravenous morphine patient-controlled anesthesia. The authors believe that this lack of consensus stems from varied epidural dosing protocols and techniques in catheter placement, which they have witnessed anecdotally at their own institution. This prompted the authors to develop and refine a standardized dosing and catheter placement protocol for pain control after spinal surgery. METHODS: Epidural catheters were placed intraoperatively before wound closure, then removed on postoperative Day 5. Dosing consisted of fentanyl (1 microg/kg) and hydromorphone (5 microg/kg) diluted in preservative-free saline (0.2 mL/kg). After surgery, dosing consisted of 0.1% ropivacaine and hydromorphone (10 microg/ml) continuously infused at 0.2 mL/kg/hour. Postoperative pain control was assessed on each postoperative day using a visual analog pain scale with choices ranging from 0 to 10. RESULTS: The arithmetic mean of the median pain scores after surgery was 2.1. The mean of the maximum pain scores for the 5 days was 4.1. Three patients required an epidural bolus and a 20% increase in the epidural infusion rate. One patient was judged to be excessively sleepy, so the epidural infusion rate was decreased by 20%. Pruritus requiring diphenhydramine developed in three patients. No other adverse effects related to epidural analgesia were noted. No catheters were accidentally pulled out or disconnected. CONCLUSION: By following the dosing protocol described, epidural catheters can be used safely and effectively to control postoperative pain after anterior instrumentation and spinal fusion for adolescent scoliosis. PMID- 11389402 TI - Traumatic paraplegia in snowboarders. AB - STUDY DESIGN: Patients with traumatic paraplegia resulting from snowboarding accidents were reviewed. OBJECTIVE: To understand the clinical features and mechanisms of paraplegic snowboarding injuries. SUMMARY OF BACKGROUND DATA: The recent explosion in the popularity of snowboarding has resulted in dramatically increased numbers of snowboarding injuries. However, little information is available as to the types and mechanisms of snowboard-related spinal injuries and their neurologic involvement. METHODS: The subjects of this study were six male patients, with an average age of 23.7 years, referred to the authors' institution for neurologic deficits associated with spinal injuries between January 1996 and March 1999. The clinical features of these patients were reviewed with respect to the mechanism of the injury, fracture pattern, neurologic status, treatment, and clinical outcome. The mean follow-up period was 23.7 months. RESULTS: The six snowboarders with traumatic paraplegia constituted a very homogenous group with the following features: They were all young men between the ages of 23 and 25 years. All the injuries had occurred at the vertebral junctions. The primary mechanism of the fractures was a backward fall from an intentional jump. The fracture patterns were of the flexion-distraction type. These homologous features suggest that this snowboarding group is at high risk for severe spinal injury. CONCLUSION: The high risk of traumatic paraplegia for a group within the snowboarding population requires the development and provision of injury prevention strategies specific to this group. PMID- 11389403 TI - Imaging corner. Unknown case. Spinal MRI after head-on collision. PMID- 11389404 TI - Rotatory fixation. PMID- 11389405 TI - Prediction of progression of spinal deformity in Duchenne muscular dystrophy: a preliminary report. AB - STUDY DESIGN: Discriminatory power was statistically estimated for multiple combinations of risk indicators for the progression of spinal deformity in Duchenne muscular dystrophy (DMD). OBJECTIVE: To differentiate DMD cases in which spinal deformity will rapidly and severely progress from those with lesser progression of spinal deformity. SUMMARY OF BACKGROUND DATA: Early surgical intervention using instrumentation has recently been advocated for DMD patients to prevent the progression of spinal deformity. However, early determination of cases needing surgical intervention is difficult because of variations in the severity of the clinical courses of DMD patients. METHODS: Charts and spinal radiographs of 12 DMD patients were reviewed retrospectively. Patterns of progression in spinal deformity were classified into three types according to Oda's classification. Discriminant analysis was conducted to categorize the patients into either a severe progression group (type-1 and type-2 patients) or a less severe progression group (type-3 patients and patients without spinal deformity) on the basis of four predictors: 1) vital capacity at the age of 10 years, 2) the age at which ambulation ceased, 3) curve pattern of spinal scoliosis, and 4) Cobb angles of spinal scoliosis at the age of 10 years. RESULTS: Eleven of the twelve DMD patients showed spinal deformity. Three were classified as type 1, six were classified as type 2, and two were classified as type 3. The remaining patient showed no spinal deformity. Multiple discriminant analysis correctly predicted the severity of the clinical course of 91.7% of the DMD patients. Vital capacity at age 10 was found to be the strongest predictor among the variables. CONCLUSIONS: Through multiple discriminant analysis, the clinical course of spinal deformity in DMD patients was correctly predicted in 92% of subjects. This method would be useful to determine early which DMD cases need surgical intervention for treatment of spinal deformity. PMID- 11389406 TI - Reliability of centroid, Cobb, and Harrison posterior tangent methods: which to choose for analysis of thoracic kyphosis. AB - STUDY DESIGN: Thirty lateral thoracic radiographs were digitized twice by each of the three examiners. OBJECTIVES: To determine the reliability of the centroid, Cobb, and Harrison posterior tangent methods when applied to analysis of thoracic kyphosis. BACKGROUND DATA: Reliability studies on measurements of thoracic kyphosis are rare. METHODS: Blind, repeated-measures design was used. Thirty lateral thoracic radiographs were digitized twice by each of three examiners. To evaluate reliability of determining global and segmental alignment, vertebral bodies of T1-T12 were digitized. Centroids at the intersection of vertebral body diagonals and tangents to posterior vertebral bodies were constructed by computer. Also the computer constructed global and segmental centroid angles, Cobb angles (two-line method), and posterior tangent intersection angles from T1 to T12. Interclass and Intraclass correlation coefficients for these data were calculated and interpreted. RESULTS: From the points selected by examiners, all three methods have similar high ICC values for the global angles (> 0.94). For the segmental angles, the interobserver and intraobserver reliability is also very similar for all three methods, with ICCs in the good and excellent ranges (0.59-0.75 and 0.75-1.0, respectively). The mean absolute differences of observers' measurements are low, similar, and in the range of 0.9 degrees to 2.5 degrees. CONCLUSIONS: The centroid, two-line Cobb, and Harrison posterior tangent methods, when applied to measurements of kyphosis, are all reliable and have similar small error ranges. The centroid method does not give an accurate segmental analysis, uses more points and more time in clinical applications, and results in smaller angles of total kyphosis than the Cobb or posterior tangent methods. The posterior tangents are the slopes along the curve. PMID- 11389407 TI - Radiographic analysis of lumbar lordosis: centroid, Cobb, TRALL, and Harrison posterior tangent methods. AB - STUDY DESIGN: Delayed, repeated measures, with three examiners each twice digitizing thirty lateral lumbar radiographs. OBJECTIVES: To determine the reliability and clinical utility of the centroid, Cobb, tangential radiologic assessment of lumbar lordosis (TRALL), and Harrison posterior tangent line drawing methods for analysis of lumbar lordosis. BACKGROUND DATA: Cobb's method is commonly used for curvature analysis on lateral lumbar radiographs, whereas the centroid, TRALL, and Harrison posterior tangent methods are not widely used. METHODS: Thirty lateral lumbar radiographs were digitized twice by each of three examiners. To evaluate reliability of determining global and segmental alignment, all four vertebral body corners of T12-S1 and the superior margin of the femur head were digitized. Angles created were segmental and global centroid, (two line) Cobb angles, and intersections of posterior tangents. A global TRALL angle was determined. Means, standard deviations, mean absolute differences, interclass and intraclass correlation coefficients (ICC), and confidence intervals were calculated. RESULTS: The interobserver and intraobserver reliabilities of measuring all segmental and global angles were in the high range (ICCs > 0.83). The mean absolute differences of observers' measurements were small (0.6 degrees 2.0 degrees ). Distal segmental (L4-S1) and global angles of lumbar curvature were dependent on the method of measurement. CONCLUSIONS: All four radiographic methods had high reliability and low mean absolute differences of observers' measurements. Because it lacks a segmental analysis, the TRALL method is not recommended. The centroid, Cobb, and Harrison posterior tangent methods provide global and segmental angles. However, the centroid segmental method requires three segments and is less useful for a stability analysis. PMID- 11389408 TI - Long-term effects of specific stabilizing exercises for first-episode low back pain. AB - STUDY DESIGN: A randomized clinical trial with 1-year and 3-year telephone questionnaire follow-ups. OBJECTIVE: To report a specific exercise intervention's long-term effects on recurrence rates in acute, first-episode low back pain patients. SUMMARY OF BACKGROUND DATA: The pain and disability associated with an initial episode of acute low back pain (LBP) is known to resolve spontaneously in the short-term in the majority of cases. However, the recurrence rate is high, and recurrent disabling episodes remain one of the most costly problems in LBP. A deficit in the multifidus muscle has been identified in acute LBP patients, and does not resolve spontaneously on resolution of painful symptoms and resumption of normal activity. Any relation between this deficit and recurrence rate was investigated in the long-term. METHODS: Thirty-nine patients with acute, first episode LBP were medically managed and randomly allocated to either a control group or specific exercise group. Medical management included advice and use of medications. Intervention consisted of exercises aimed at rehabilitating the multifidus in cocontraction with the transversus abdominis muscle. One year and three years after treatment, telephone questionnaires were conducted with patients. RESULTS: Questionnaire results revealed that patients from the specific exercise group experienced fewer recurrences of LBP than patients from the control group. One year after treatment, specific exercise group recurrence was 30%, and control group recurrence was 84% (P < 0.001). Two to three years after treatment, specific exercise group recurrence was 35%, and control group recurrence was 75% (P < 0.01). CONCLUSION: Long-term results suggest that specific exercise therapy in addition to medical management and resumption of normal activity may be more effective in reducing low back pain recurrences than medical management and normal activity alone. PMID- 11389409 TI - Recurrent calcified spinal meningioma detected by plain radiograph. AB - STUDY DESIGN: A case report of a patient presenting with a rare calcified spinal meningioma that recurred and became symptomatic 33 years after initial surgery was performed. OBJECTIVES: To demonstrate a calcified recurrent spinal meningioma visible on a plain radiograph. The present case is discussed in conjunction with a review of previous cases of calcified spinal meningioma. SUMMARY OF BACKGROUND DATA: Meningioma is a common spinal tumor that has a favorable prognosis if resected completely. Microscopic calcification is frequently found in spinal meningiomas because of psammoma bodies. However, calcified meningiomas in the spinal canal that can be detected on a plain radiograph are uncommon. To the authors' knowledge, there have been no previous reports of recurrent spinal meningioma with calcification visible on a plain radiograph. METHODS: A patient experienced difficulty in walking and was diagnosed as having a spinal meningioma. Laminectomies and a total tumor excision were performed. Histologically, the tumor was diagnosed as a psammomatous meningioma. The tumor recurred and became symptomatic 33 years after the initial operation. The tumor was detected on a plain radiograph, appearing as a calcified tumor at the same thoracic level as the site of the initial operation. The recurrent tumor was completely removed, and a histologic examination showed huge psammoma bodies with calcification, but without ossification. RESULTS: Preoperative paraplegia improved after the surgery, and she was able to walk with a crutch. The patient was discharged from the hospital. CONCLUSIONS: The recurrent psammomatous meningioma contained clusters of calcified psammoma bodies, resulting in a hard tumor that was visible on a plain radiograph. PMID- 11389410 TI - Presidential address: what would you do if it were your father? Reflections on endovascular abdominal aortic aneurysm repair. PMID- 11389411 TI - Thrombolysis for native arterial occlusions of the lower extremities: clinical outcome and cost. AB - INTRODUCTION: Intra-arterial thrombolysis is commonly used as the initial treatment of acute or subacute lower extremity ischemia. METHODS: To evaluate the efficacy and cost of thrombolysis, we retrospectively analyzed 100 consecutive cases (87 patients) in which intra-arterial lysis (urokinase) was used as the initial treatment for native arterial lower extremity occlusive disease. The mean age of patients was 67 years, 57% of the patients were male, and preexisting peripheral vascular disease was present in 74%. Presenting symptoms were limb threatening ischemia (53%) and claudication (47%). Acute symptoms (< 2 weeks' duration) were present in 48%. RESULTS: The 30-day morbidity rate was 31%, and four patients died. Complications were significant bleeding (23%), ischemic stroke (1%), and renal failure with (2%) and without (2%) dialysis. Concomitant angioplasty was performed in 63%. Complete or significant lysis as demonstrated with angiography was achieved in 75% of iliac, 58% of femoropopliteal, and 41% of crural vessels (P <.001). Within 30 days of lysis, 9% of patients underwent major amputation and 20% surgical revascularization (in 3 patients the extent of revascularization was lessened by the lytic therapy). Amputation-free survival was 83% and 75% at 6 months and 2 years, respectively. Relief of ischemia (defined as relief of claudication or limb salvage without major surgical intervention) was achieved in only 70% and 43% of patients at 30 days and 2 years, respectively (Kaplan-Meier analysis; mean follow-up, 31 months). Patients with aortoiliac disease had significantly better outcomes than those with infrainguinal disease (P =.03). Duration or type of presenting symptoms did not predict outcome. The cost of the initial hospitalization per patient for thrombolysis was $18,490. CONCLUSION: Thrombolysis can be as or more costly than surgery and is associated with a suboptimal outcome in a significant number of patients. These data lead us to caution against a uniform policy of initial thrombolysis for patients who present with lower extremity ischemia. PMID- 11389412 TI - Factors influencing ulcer healing in patients with combined arterial and venous insufficiency. AB - PURPOSE: The purpose of this study was to determine the effectiveness of treatment of patients with combined arterial and venous insufficiency (CAVI), evaluate variables associated with successful ulcer healing, and better define criteria for interventional therapy. STUDY DESIGN: We retrospectively reviewed the records of patients treated at four institutions from 1995 to 2000 with lower extremity ulcers and CAVI. Arterial disease was defined as an ankle/brachial index less than 0.9, absent pedal pulse, and at least one in-line arterial stenosis > 50% by arteriography. Venous insufficiency was defined as characteristic clinical findings and duplex findings of either reflux or thrombus in the deep or superficial system. Clinical, demographic, and hemodynamic parameters were statistically analyzed with multiple regression analysis and correlated with ulcer healing and limb salvage. RESULTS: Fifty-nine patients with CAVI were treated for nonhealing ulcers that had been present from 1 to 39 months (mean, 6.4 months). All patients had edema. The mean ankle/brachial index was 0.55 (range, 0-0.86). Treatment included elastic compression and leg elevation in all patients and greater saphenous vein stripping in patients with superficial venous reflux. Fifty-two patients underwent arterial bypass grafting, three underwent an endarterectomy, one underwent superficial femoral artery percutaneous transluminal angioplasty, and three underwent primary below-knee amputation. For purposes of analysis, patients were divided into four groups according to the pattern of arterial and venous disease and the success of arterial reconstruction. Group 1 consisted of 22 patients with a patent arterial graft, superficial venous incompetence, and normal deep veins. Group 2 consisted of seven patients with a patent graft, superficial reflux, and deep venous reflux. Group 3 included 22 patients with a patent graft and deep venous thrombosis (DVT), and group 4 included eight patients with an occluded arterial graft. Follow-up ranged from 2 to 47 months (mean, 21.6 months). Forty-nine patients remained alive, and 10 died of unrelated causes. During follow-up, 48 of the 56 treated arteries remained patent and eight occluded. Thirty-four ulcers (58%) healed, 18 ulcers (31%) did not heal, and 7 patients (12%) required below knee amputation for nonhealed ulcers and uncontrolled infection. No patient with graft occlusion was healed, and 12 ulcers persisted despite successful arterial reconstruction. Twenty-one (78%) of 27 patients undergoing greater saphenous vein stripping were healed, but none of these patients had DVT. The mean interval from bypass graft to healing was 7.9 months. Thirty-two (68%) of 46 patients without prior DVT were healed, whereas only two (15%) of 13 patients with prior DVT were healed, and this variable, in addition to graft patency, was the only factor statistically significant in predicting healing (P <.05). CONCLUSIONS: Ulcers may develop anywhere on the calf or foot in patients with CAVI, and healing requires correction of arterial insufficiency. Patients with prior DVT are unlikely to heal, even with a patent bypass graft. Ulcer healing is a lengthy process and requires aggressive treatment of edema and infection, and successful arterial reconstruction. Patients with a prior DVT are unlikely to benefit from aggressive arterial or venous reconstruction. PMID- 11389413 TI - Early results of infragenicular revascularization based solely on duplex arteriography. AB - PURPOSE: Recent reports have both advocated and questioned the utility of duplex arteriography (DA) as the sole preoperative imaging modality for planning infragenicular revascularization. This study compares the outcome of patients with critical limb ischemia who underwent infragenicular vein grafts on the basis of DA alone versus conventional preoperative contrast arteriography (CA). METHODS: The study group is composed of 23 consecutive patients who underwent infragenicular vein bypass grafting solely on the basis of preoperative DA from 1998 to 1999. They were compared with 50 consecutive patients who underwent infragenicular vein bypass grafting after CA from 1996 to 1998. Peak systolic velocity and end-diastolic velocity of potential target arteries were recorded during DA studies. In situ saphenous vein grafts were used preferentially, and technical adequacy of all grafts was assessed with completion duplex or arteriography. RESULTS: DA and CA groups were comparable on the basis of age and risk factors. In one limb (4%), the target artery selected by DA was abandoned because of dense calcification. No other revision in target or inflow artery was required on the basis of intraoperative completion studies. At 1 year, primary graft patency (78% vs 70%, P =.72) and limb salvage (70% vs 81%, P =.21) were comparable between the two groups. In the DA group, mean preoperative target artery peak systolic velocity in patent versus failed grafts was 49 +/- 18 cm/s versus 31 +/- 9 cm/s (P =.04), whereas mean end-diastolic velocity was 22 +/- 7 cm/s versus 14 +/- 8 cm/s (P =.08). CONCLUSION: Infragenicular revascularization directed by DA alone provides early graft patency and limb salvage rates comparable to similar procedures that are based on CA. Preoperative DA target artery velocities may predict outcome and improve target selection. These initial results justify further clinical testing of DA as the primary imaging modality for planning infragenicular vein grafts. PMID- 11389414 TI - Infrainguinal bypass grafting in patients with end-stage renal disease: improving outcomes? AB - OBJECTIVE: This study was undertaken to examine recent trends in the outcomes of patients with end-stage renal disease (ESRD) undergoing infrainguinal bypass grafting (IBG) with autogenous vein. METHODS: A retrospective analysis of all IBGs performed on patients with ESRD at a single tertiary care institution during the interval 1993 to 1999 was undertaken. The comparison groups consisted of concurrent series of patients with elevated creatinine (creatinine level > 1.2 mg/dL) and patients with normal renal function undergoing IBG. Procedural variables, angiographic runoff scores, and extent of tissue necrosis at presentation were correlated with outcome. Categoric parameters were compared with chi(2) analysis; rates were computed with life-table analysis. RESULTS: Of an overall cohort of 622 IBGs performed during this interval, 78 IBGs (12.5%) were performed on 60 patients with ESRD, with a perioperative mortality rate of 1.3% that was comparable to controls. All reconstructions in the ESRD cohort were for limb salvage indications. Four-year survival, primary, assisted primary, and secondary patency rates for the ESRD group were 51% +/- 9%, 60% +/- 11%, 86% +/- 5%, and 86% +/- 5%, respectively; these were not statistically different from the control groups. Limb salvage in the ESRD group was 77% +/- 6% at 4 years and was significantly less then either the elevated creatinine (92% +/- 4%; P <.02) or the normal renal function group (90% +/- 2%: P <.02). Of 16 amputations in the ESRD group, nine were performed in limbs with patent grafts. The only absolute predictor of limb loss despite a patent graft was the presence of a heel ulcer more than 4 cm in diameter. Age, runoff score of the International Society for Cardiovascular Surgery/Society for Vascular Surgery, isolated tibial bypass graft, and location of distal anastomosis were not predictive of hemodynamic failure. CONCLUSIONS: Patients with ESRD constitute an increasing proportion of patients undergoing IBG in a tertiary care setting. Four-year survival, perioperative mortality, and graft patency rates are similar to patients with normal renal function and support an aggressive approach to this population. Major limb amputation despite a patent graft remains a problem of unique frequency in patients with ESRD. Adequate predictors of hemodynamic failure of IBG in this group do not exist, although a heel ulcer more than 4 cm may indicate an unsalvageable foot. PMID- 11389415 TI - Outcome after combined carotid endarterectomy and coronary artery bypass is related to patient selection. AB - OBJECTIVE: The optimal management of patients with significant coronary and carotid artery disease remains controversial. Since reporting on a series of 100 patients undergoing combined carotid endarterectomy and coronary artery bypass (CEA/CAB) 4 years ago, we have liberalized our selection criteria for combined operation. We sought to compare outcomes of the recent cohort of 74 patients and the previous group. METHODS: All patients who underwent CEA/CAB since 1984 have been tracked in a database containing identifying information, demographic factors, anatomic information, details of surgery, and short- and long-term follow-up data. We compared the 74 patients (Group 2) undergoing CEA/CAB since 1994 with the previously reported group of 100 patients (Group 1) who underwent CEA/CAB between 1984 and 1994. We examined demographic and comorbidity factors, presence of cerebrovascular symptoms, degree of contralateral carotid stenosis, and perioperative stroke and death. Statistical comparisons were made with the chi(2) test. RESULTS: The groups had similar age and sex distributions and similar incidences of hypertension, diabetes, congestive heart failure, prior myocardial infarction, and hypercholesterolemia. More patients in Group 1 had preoperative transient cerebral ischemia or monocular blindness (55% vs 31%, P <.002) and preoperative stroke (18% vs 7%, P <.03). More patients in Group 2 had unilateral asymptomatic carotid artery stenosis (55% vs 18%, P <.001). The incidence of all perioperative strokes was higher in Group 1 (9% vs 1.4%, P <.035). There were fewer deaths (3% vs 8%) and ipsilateral strokes (0 vs 4%) in Group 2, though these were not statistically significant. CONCLUSION: We have liberalized our criteria for performing combined CEA/CAB, such that more than 50% of our recent patients have asymptomatic unilateral carotid stenosis. This practice is associated with a lower incidence of all perioperative strokes and a trend toward lower ipsilateral stroke and death. These observations suggest that perioperative stroke after CEA/CAB is related to patient selection and that low risk patients can undergo CEA/CAB with the benefits of low morbidity, patient convenience, and cost savings from avoiding a second hospitalization and operation. PMID- 11389416 TI - Contemporary management of aortic branch compromise resulting from acute aortic dissection. AB - PURPOSE: In an earlier report, we documented the incidence and impact of aortic branch compromise complicating acute aortic dissection (AD) over a 21-year interval (1965-1986). In the current study, management of peripheral vascular complications (PVCs) of AD over the past decade was reviewed. METHODS: Medical records of patients treated for AD over the interval January 1, 1990, to December 31, 1999, were reviewed. Patients with branch compromise confirmed with radiography or operation and patients with spinal cord ischemia that was based on results of a physical examination defined the study group. Comparisons between subgroups with and without PVC over a 30-year interval were analyzed with the chi(2) test. RESULTS: A total of 187 patients (101 proximal and 86 distal) were treated for AD over the study interval. A total of 53 (28%) of these patients had clinical evidence of organ or limb malperfusion (7 cerebral, 3 upper extremity, 5 spinal cord, 11 mesenteric, 12 renal, and 24 lower extremity [sites inclusive]), and one of three (17 patients) of these underwent specific peripheral vascular intervention. The remaining 65% (36) of the PVC group had complete or partial malperfusion resolution after central aortic therapy (medical or surgical) alone. Open techniques for treating PVC included aortic fenestration (9), femorofemoral grafting (2), and aortofemoral grafting (1). All had favorable outcomes with no mortality. Endovascular procedures in five patients included abdominal aortic fenestration (3) or stenting of the renal (2), mesenteric (2), and iliac (1) arteries with clinical success in three patients and two deaths. The in-hospital mortality rate for the entire group of 187 patients was 18% (15% for proximal aortic operation, 8% in medically treated patients). The presence of aortic branch compromise was not a statistically significant predictor of the patient mortality rate (23% with and 16% without; P =.26). Overall mortality rate in the current study (18% vs 37%; P =.000006) and the mortality rate with PVC (23% vs 51%; P =.001), in particular with mesenteric ischemia (36% vs 87%; P =.026), decreased significantly when compared with prior experience. CONCLUSIONS: The overall mortality rate from AD during the past decade has decreased significantly. Similar trends were noted in patients with PVCs, a previously identified high-risk subgroup. Increased awareness and prompt, specific management of PVCs, in particular when visceral ischemia is present, have contributed to improved outcomes in patients with AD. PMID- 11389417 TI - Hypogastric artery embolization in endovascular abdominal aortic aneurysm repair. AB - PURPOSE: Iliac artery anatomy is a central factor in endoluminal abdominal aortic aneurysm therapy. It serves as the conduit for graft deployment and as the region of distal graft seal. Thirty-eight percent of iliac vessels in our patients require special treatment because of aneurysms, tortuosity, or small size. Bilateral hypogastric artery exclusion has been avoided because of concerns of colorectal ischemia, hip/buttock claudication, and impotence. We suggest that elective, staged, bilateral hypogastric embolization can be performed safely with reasonably low morbidity and can expand the anatomic boundaries for stent-graft abdominal aortic aneurysm repair. METHODS: This study was performed as a retrospective chart review of patients requiring hypogastric artery embolization for endovascular repair of abdominal aortic aneurysms between June 1998 and June 2000. Patients with otherwise appropriate anatomy and common iliac artery aneurysms were informed of the option for stent-graft repair with internal iliac artery embolization with its risks of impotence, hip/buttock claudication, and bowel ischemia. Patients underwent unilateral or staged bilateral coil embolizations of their proximal hypogastric arteries with an approximate 1-week interval between procedures. Hospital and office records were reviewed; phone interviews were performed. Follow-up ranged from 1 to 12 months. RESULTS: During a 24-month period, 65 patients underwent endovascular abdominal aortic aneurysm repair; 18 patients (28%) required hypogastric artery embolization. Seven (39%) of these patients underwent bilateral embolization. There were no episodes of clinically evident bowel ischemia. Lactate levels were normal in all measured patients. Postoperative fevers (> 101.0 degrees F) were documented in 10 (56%) of 18 patients. The average white blood cell count was 12.8 x 10(9)/L (range, 8.5 22.9). There were no positive blood culture results. The return to the full preoperative diet occurred in 1 to 3 days. Hip/buttock claudication occurred in approximately 50% of patients with persistent but improved symptoms at 6 months. Eighty-seven percent of patients had preoperative erectile dysfunction. Only two patients noted worsening of erectile function postoperatively. CONCLUSIONS: Preliminary results indicate that bilateral hypogastric artery embolization can be performed, when necessary, with reasonable morbidity in patients undergoing stent-graft abdominal aortic aneurysm repair. PMID- 11389418 TI - The effect of combined arterial hemodynamics on saphenous venous endothelial nitric oxide production. AB - INTRODUCTION: Evidence exists that an ideal bypass conduit should have a functional endothelial cell surface combined with mechanical properties similar to those of native arteries. We hypothesized that the effect of combined arterial levels of pulsatile shear stress, flow, and cyclic strain would enhance saphenous venous endothelial cell nitric oxide (NO) production, and that variations in these "ideal" conditions could impair this function. We studied NO production as a measure of endothelial function in response to different hemodynamic conditions. METHODS: Human adult saphenous venous endothelial cells were cultured in 10-cm silicone tubes, similar in diameter (5 mm) and compliance (6%) to a medium-caliber peripheral artery (eg, popliteal). Tube cultures were exposed to arterial conditions: a combined pressure (120/80 mm/Hg; mean, 100 mm/Hg), flow (mean, 115 mL/min) and cyclic strain (2%), with a resultant pulsatile shear stress of 4.8 to 9.4 dyne/cm2 (mean, 7.1). Identical tube cultures were used to study variations in these conditions. Modifications of the system included a noncompliant system, a model with nonpulsatile flow, and a final group exposed to pulsatile pressure with no flow. NO levels were measured with a fluorometric nitrite assay of conditioned media collected at 0, 0.25, 0.5, 1, 2, and 4 hours. Experimental groups were compared with cells exposed to nonpulsatile, nonpressurized low flow (shear stress 0.1 dyne/cm2) and static cultures. RESULTS: All experimental groups had greater rates of NO production than cells under static conditions (P <.05). Cells exposed to ideal conditions produced the greatest levels of NO. Independent decreases in compliance, flow, and pulsatility resulted in significantly lower rates of NO production than those in the group with these conditions intact (vs noncompliant P <.05, vs nonflow P <.05, and vs nonpulsatile P <.05). CONCLUSIONS: Our results show that in the absence of physiologically normal pulsatility, cyclic strain, and volume flow, endothelial NO production does not reach the levels seen under ideal conditions. Pulsatile flow and compliance (producing flow with cyclic stretch) play a key role in NO production by vascular endothelium in a three-dimensional hemodynamically active model. This correlates biologically with clinical experience linking graft inflow and runoff and the mechanical properties of the conduit to long-term patency. PMID- 11389419 TI - Senescence and the healing rates of venous ulcers. AB - OBJECTIVE: Premature cellular senescence has been linked to venous hypertension and may contribute to delayed healing of venous ulcers. We hypothesized that the percentage of senescent cells in in vitro populations of fibroblasts isolated from venous ulcers is directly related to the clinical time-to-healing. METHODS: Biopsy specimens were obtained from ulcer margins and unaffected dermal tissue of the ipsilateral thigh of seven patients with active venous ulcers. Using explant culture techniques, we obtained populations of wound fibroblasts and normal fibroblasts. The percentage of senescence in these cell populations was determined with X-Gal (5-bromo-4-chloro-3-indolyl beta-D-galactoside), which was used as a stain for B-galactosidase, a biomarker for senescent dermal fibroblasts. The X-Gal stain is a peroxidase stain for B-galactosidase. All patients in the study were treated with compression dressings. On a weekly basis, digital images were taken until ulcers healed. Planimetric healing rates were calculated from these images, and an overall time-to-healing was recorded. All cytologic investigations were performed on first passage cells. RESULTS: The average starting ulcer size was 4.2 cm2. Five of the data points represented healed ulcers. The two remaining patients withdrew from the study to pursue other therapies after having been treated with compression dressings for a long time. Linear regression analysis of healed ulcers identified a relationship between percent of senescence and time-to-healing, which was statistically significant (R2 = 0.81, P =.037). High percentages of senescent cells also had a correlation with slowed planimetric healing, which was not statistically significant. CONCLUSIONS: This study demonstrates a clinical correlation between quantitative in vitro senescence and time-to-healing. A percentage of senescence that is greater than 15% in populations of cells isolated from venous ulcers may identify a "difficult to heal" ulcer. There is no good clinical indicator for determining the likelihood of ulcer healing, but these results indicate that senescence percentage may have potential in this regard. PMID- 11389420 TI - Lower extremity arterial injury: results of 550 cases and review of risk factors associated with limb loss. AB - PURPOSE: We sought to analyze the results of lower limb arterial injury (LLAI) management in a busy metropolitan vascular unit and to identify risk factors associated with limb loss. PATIENTS AND METHODS: Between 1987 and 1997, prospectively collected data on 550 patients with 641 lower limb arterial injuries were analyzed. RESULTS: The mechanism of LLAI was gunshot wounds in 46.1%, blunt in 19%, stabbing in 11.8%, and shotgun in 9.1%. The most frequently injured vessel was the superficial femoral artery (37.2%), followed by the popliteal (30.7%), crural (11%), common femoral (8.7%), and deep femoral (5.3%) arteries. In 3.4% of cases, there was a combined injury on either side of the knee (ipsilaterally). Associated injuries included bony injury in 35.1% of cases, nerve injury in 7.6%, and remote affecting the head, chest, or abdomen in 3.6%. Surgery was carried out on 96.2% of cases with a limb salvage rate of 83.8% and a survival of 98.5%. In spite of a rising trend in LLAI, our total and delayed amputation rates remained stable. On stepwise logistic regression analysis, significant (P <.01) independent risk factors for amputation were occluded graft (odds ratio [OR] 16.7), combined above- and below-knee injury (OR 4.4), tense compartment (OR 4.2), arterial transsection (OR 2.8), and associated compound fracture (OR 2.7). CONCLUSION: LLAI carries a high amputation rate. Stab injuries are the least likely to lead to amputations, whereas high-velocity firearm injuries are the most likely to do so. The most significant independent risk factor for limb loss was failed revascularization. PMID- 11389421 TI - Outcomes after surgery for thoracic outlet syndrome. AB - PURPOSE: This study determined whether there is an association between psychological and socioeconomic characteristics and the long-term outcome of operative treatment for patients with sensory neurogenic thoracic outlet syndrome (N-TOS). METHODS: Clinical records, preoperative psychological testing results, and long-term follow-up questionnaire data were reviewed for consecutive patients who underwent surgery for N-TOS from 1990 to 1999. Multivariate logistic regression models were developed as a means of identifying independent risk factors for postoperative disability. RESULTS: Operative decompression of the brachial plexus via a supraclavicular approach was performed for upper extremity pain and paresthesia with no mortality and minimal morbidity in 170 patients. After an average follow-up period of 47 months, 65% of patients reported improved symptoms, and 64% of patients were satisfied with their operative outcome. However, 35% of patients remained on medication, and 18% of patients were disabled. Preoperative factors associated with persistent disability include major depression (odds ratio [OR], 15.7; P =.02), not being married (OR, 7.9; P =.04), and having less than a high school education (OR, 8.1; P =.09). CONCLUSION: Operative decompression was beneficial for most patients. Psychological and social factors, including depression, marital status, and education, are associated with self-reported disability. The impact of the preoperative treatment of depression on the outcome of TOS decompression should be studied prospectively. PMID- 11389422 TI - Endovascular repair of aortic aneurysms: critical events and adjunctive procedures. AB - OBJECTIVE: We sought to define the learning curve relative to the incidence and range of intraoperative problems and to establish guidelines for troubleshooting during the endovascular repair of infrarenal aortic aneurysms. METHODS: We prospectively evaluated our first 75 consecutive cases over a 12-month period and focused on perioperative critical events and adjunctive procedures as categorical outcome measures collected during the operation. Patients were separated into three groups on the basis of the date of their operation, such that group 1 consisted of our first 25 cases, group 2 our next 25 cases, and group 3 our last 25 cases. RESULTS: At least one critical event and adjunctive procedure marked 67 (89%) of 75 cases. In 51%, there were at least two critical events and adjunctive procedures. There were no immediate open conversions or intraoperative deaths. Access problems occurred in 28% of the 75 cases and were addressed by use of brachial-femoral artery access (30%), iliac artery/aortic bifurcation balloon angioplasty (8%), and iliofemoral conduits (4%). Graft foreshortening was the most common deployment event (44%), necessitating distal covered extensions. Iliac graft limb twists and kinks occurred in 12% of cases and were managed with balloon angioplasty and uncovered stents. General incidents included balloon ruptures (10%), arterial dissections (6%), iliac artery rupture (2.6%), and lower extremity ischemia (4%). The two cases of iliac artery rupture were managed with distal covered extensions, and there were no cases of atheroemboli. Intraoperative endoleaks were encountered in 44% of the cases and included proximal attachment sites (15%), distal attachment sites (9%), type 2 sources, and "blushes." Management of intraoperative endoleaks included proximal/distal covered extensions and re-ballooning. Our 30-day endoleak rate was 20%. The incidence of critical events did not decrease in the latter one third compared with the first two thirds of cases. CONCLUSIONS: Critical events occur frequently during endovascular repair of aortic aneurysms. The intraoperative problems range from the common endoleaks, access and deployment issues, and balloon ruptures, to rare but life-threatening complications such as iliac artery rupture. A toolbox of accessories that includes wires, catheters, large balloons, covered proximal and distal extensions, and uncovered stents is essential given the frequency of adjunctive procedures. Successful aortic endografting requires more than mere familiarity with basic endovascular techniques. PMID- 11389423 TI - Changes in cellular motility and cytoskeletal actin in fibroblasts from patients with chronic venous insufficiency and in neonatal fibroblasts in the presence of chronic wound fluid. AB - PURPOSE: Fibroblasts (fb) play an important role in wound healing involving motility, contraction, fibrosis, and expression of the cytoskeletal protein alpha smooth muscle actin (alpha-sma). Patients with chronic venous insufficiency (CVI) are known to have dermal changes and impaired venous ulcer healing. To investigate whether these dermal-fb have an altered ability to migrate and whether chronic wound fluid from venous ulcers alters neonatal fb motility, we examined cell migration and alpha-sma. METHODS: Fibroblasts were cultured from the margin of venous ulcers (du-fb, n = 4, CEAP 6), from patients with venous reflux without ulcer (dr-fb, n = 5, CEAP 2), and from the ipsilateral thigh of the same patients with (pu-fb) and without (pr-fb) ulcer, respectively. The abbreviations used are p and d, which represent proximal and distal, respectively; u and r represent ulcer and reflux, respectively. Neonatal foreskin fibroblasts (nf-fb) were exposed to chronic venous ulcer wound fluid (CVUWF, 300 microg protein/mL, n = 3) or bovine serum albumin (BSA, control). Fibroblast motility was determined by means of time-lapse photo-images, and the rate (micrometer per hour) was calculated. Immunohistochemistry for alpha-sma was analyzed with confocal laser microscopy. RESULTS: The rate of motility (micrometer per hour +/- SEM) was decreased for both du-fb (11.4 +/- 0.7) and dr fb (13.8 +/- 0.6), when compared with pu-fb (21.9 +/- 0.9) and pr-fb (24.7 +/- 1.1), respectively. The motility rate for nf-fb was lower in CVUWF (24.7 +/- 2.0) than in BSA (37.1 +/- 6.7). An elevated level of microfilament bundles of alpha sma for both du-fb and dr-fb, compared with those of pu-fb and pr-fb, and also in nf-fb treated with CVUWF was demonstrated by means of immunohistochemistry. CONCLUSION: These data demonstrate a reduced motility in the dermal fb of patients with CVI. Patients with reflux disease without ulcer are predisposed to these changes. Furthermore, it appears that CVUWF causes changes in motility and alpha-sma expression in nf-fb as demonstrated in du-fb. These findings suggest that reduced motility and CVUWF, representing the microenvironment of venous ulcers, play a significant role in impaired wound healing. PMID- 11389424 TI - Plasma endothelin levels and outcome in patients undergoing repair of ruptured infrarenal abdominal aortic aneurysm. AB - BACKGROUND: Endothelin-1 (ET-1) is the most potent known vasoconstrictor. Elevated plasma levels have been demonstrated in patients with myocardial infarction, cardiogenic and septic shock, and respiratory, heart, and kidney failure, as well as in those undergoing elective abdominal aortic aneurysm (AAA) repair. However, endothelin levels have not previously been examined in patients undergoing repair of ruptured AAA. We hypothesized that hemorrhagic shock, lower torso ischemia, and reperfusion associated with ruptured AAA repair lead to increased synthesis and secretion of ET-1, which, in turn, predispose to organ failure, one of the principal causes of death in this condition. METHODS: Fourteen patients were studied. Plasma levels of big ET-1 and ET-1 were measured immediately before operation and immediately before, 5 minutes, and 6 hours after aortic clamp release. RESULTS: All patients survived for at least 24 hours after operation. Big ET-1 levels were above the normal range at one or more sample points in all patients, and the ET-1 levels were above the normal range in all survivors and four of five nonsurvivors. Five patients who died of organ failure had significantly lower big ET-1 levels at all sample points and significantly lower ET-1 levels after 5 minutes of reperfusion when compared with survivors. Preoperative ET-1 levels were significantly lower in eight patients who subsequently developed kidney failure than in six patients who did not. CONCLUSION: Contrary to our original hypothesis, these novel data demonstrate that patients with ruptured AAA in whom fatal postoperative organ failure develops have significantly lower perioperative endothelin levels than survivors. PMID- 11389425 TI - Monocyte adhesion to balloon-injured arteries: the influence of endothelial cell seeding. AB - OBJECTIVE: Deendothelialization of injuries of the artery disrupts normal vascular homeostasis, affecting both the structural integrity of the blood vessel wall, as well as the interaction of the arterial surface with blood components such as platelets, leukocytes, and circulating proteins. Leukocyte and, in particular, monocyte recruitment to damaged vessels has been implicated in the pathogenesis of intimal hyperplasia. We hypothesize that reendothelialization is an important modulator of monocyte adhesion to healing arterial surfaces. METHODS: New Zealand white rabbits (n = 20) were subjected to bilateral iliofemoral artery balloon injury. Cultured, autologous venous endothelial cells (ECs) were immediately seeded onto one vessel, whereas the contralateral artery received medium alone, to accelerate endothelial relining. Vessels were harvested (5-9 days after injury) for analysis of permeability (Evans Blue dye exclusion), endothelial coverage (anti-CD31 immunohistochemistry), monocyte adhesion (ex vivo binding of 51Na2CrO4-labeled monocytic THP-1 cells), and monocyte recruitment (RAM-11 immunohistochemistry). RESULTS: Improved EC coverage was evidenced by positive staining for CD31 in the seeded vessels. Vessel wall permeability was markedly reduced in EC-seeded arteries (29% +/- 10% vs 99% +/- 0% surface Evans blue staining, P <.005), consistent with restoration of a functional endothelial barrier. EC seeding significantly reduced ex vivo THP-1 binding to vessels explanted at a mean of 8 days after injury (45,170 +/- 8939 vs 85,994 +/- 16,500 cells/cm2, P <.05). However, RAM-11 staining revealed no significant difference in overall macrophage accumulation between seeded and control vessels 1 week after injury (111 +/- 22 vs 95 +/- 14 cells/section, P =.36). CONCLUSIONS: Immediate seeding of a balloon-injured rabbit artery with cultured ECs results in accelerated restoration of the endothelial lining. At 1 week, barrier function is improved, and the seeded vessel surface is less adhesive to activated monocytes ex vivo, as compared with injured controls. Nonetheless, EC-seeded and nonseeded arteries demonstrate similar total macrophage accumulation over 1 week. These data suggest that after mechanical arterial injury, endothelial coverage may be one important variable influencing leukocyte adhesion. PMID- 11389426 TI - Elastase is not sufficient to induce experimental abdominal aortic aneurysms. AB - PURPOSE: Research investigating abdominal aortic aneurysms (AAAs) commonly uses a rat model dependent on aortic infusion of porcine pancreatic elastase to initiate AAA formation. Unfortunately, the sizes of AAAs generated by this model have varied widely among published studies. This may reflect lot-to-lot variations in commercial elastase preparations. This study was undertaken to investigate the ability of different lots of elastase to induce AAAs and explain the variability identified. METHODS: Four lots of elastase were evaluated in the standard rat AAA model. Saline solution was used as a control. Additional groups of rats were treated with higher concentrations of elastase with or without the macrophage activator thioglycollate medium. Aortic diameters were measured in all rats. Inflammation and elastin degradation was examined histologically. Elastase activity and purity were evaluated for all lots. RESULTS: Of the four lots tested, only one was able to consistently generate AAAs at the standard dose (P <.05). Increasing the amount of elastase infused produced AAAs in some ineffective lots. Infusion of thioglycollate medium in combination with otherwise ineffective elastase produced AAAs (P =.02). However, the elastase with the highest purity failed to generate AAAs, even at the highest dose tested or in combination with thyioglycollate medium. Thioglycollate medium alone failed to result in AAA formation. All elastase lots displayed elastolytic activity in vitro and produced elastin degradation in vivo. Elastin degradation did not correlate with AAA size in elastase-treated rats (P = NS). Aneurysm size correlated with extent of inflammation (P =.005). CONCLUSION: Induction of AAAs does not correlate with elastolytic activity. Infusion of pure elastase alone is not sufficient to induce AAA formation in spite of evidence of elastin degradation. Presumed inflammatory modifiers, which contaminate some elastase preparations, enhance AAA formation. Future use of this rat model will need to take the variability of elastase preparations into account with controls for each new elastase lot. PMID- 11389427 TI - The role of type I collagen in aortic wall strength with a homotrimeric. AB - PURPOSE: Elastin and collagen (types I and III) are the primary load-bearing elements in aortic tissue. Deficiencies and derangements in elastin and type III collagen have been associated with the development of aneurysmal disease. However, the role of type I collagen is less well defined. The purpose of this study was to define the role of type I collagen in maintaining biomechanical integrity in the thoracic aorta, with a mouse model that produces homotrimeric type I collagen [alpha1(I)]3, rather than the normally present heterotrimeric [alpha1(I)]2 alpha2(I) type I collagen isotype. METHODS: Ascending and descending thoracic aortas from homozygous (oim/oim ), heterozygous (oim /+), and wildtype (+/+) mice were harvested. Circumferential and longitudinal load-extension curves were used as a means of determining maximum breaking strength (Fmax) and incremental elastic modulus (IEM). Histologic analyses and hydroxyproline assays were performed as a means of determining collagen organization and content. RESULTS: Circumferentially, the ascending and descending aortas of oim /oim mice demonstrated significantly reduced Fmax, with an Fmax of only 60% and 23%, respectively, of wildtype mice aortas. Oim/oim descending aortas demonstrated significantly greater compliance (decreased IEM), and the ascending aortas also exhibited a trend toward increased compliance. Reduced breaking strength was also demonstrated with longitudinal extension of the descending aorta. CONCLUSION: The presence of homotrimeric type I collagen isotype (absence of alpha2(I) collagen) significantly weakens the aorta. This study demonstrates the integral role of type I collagen in the biomechanical and functional properties of the aorta and may help to elucidate the role of collagen in the development of aneurysmal aortic disease or dissection. PMID- 11389428 TI - Hypogastric artery aneurysm rupture after endovascular graft exclusion with shrinkage of the aneurysm: significance of endotension from a "virtual," or thrombosed type II endoleak. AB - Type II endoleaks, resulting from retrograde branch flow, after endovascular graft aneurysm exclusion are considered benign because they usually thrombose and are commonly associated with stable or shrinking aneurysm sacs. We report a hypogastric artery aneurysm rupture from endotension from an undetected, thrombosed Type II endoleak, associated with sac shrinkage. The patient had undergone an endovascular graft repair of a 4-cm right common iliac artery and 9 cm hypogastric artery aneurysm with distal hypogastric artery coil embolization. Serial computed tomography scans revealed no endoleak and a hypogastric aneurysm thrombosis with shrinkage. Eighteen months later, the aneurysm ruptured as a result of pressurization from backbleeding, patent branches. PMID- 11389429 TI - Calciphylaxis: a poor prognostic indicator for limb salvage. AB - INTRODUCTION: Calciphylaxis (calcific uremic arteriolopathy) is a rare complication of end-stage renal disease in patients with secondary hyperparathyroidism. Subcutaneous skin nodules and extensive tissue necrosis with vascular calcification characterize this process. DESIGN AND SETTING: The design of the study included case series and literature review. The study was set in a tertiary care hospital. PATIENTS: Four patients presented over an 8-month period with extensive tissue loss and the subsequent diagnosis of calciphylaxis. Two of these patients were revascularized. One underwent primary amputation, and the final patient died before revascularization. The mortality rate for this group was 75%. All patients had significant vascular complications outside the area of initial presentation. CONCLUSIONS: These individuals represent a subset of patients who may not benefit from revascularization or may require extensive regulation of divalent metabolism before consideration for revascularization. PMID- 11389430 TI - Successful management of tracheo-innominate artery fistula with endovascular stent graft repair. AB - Tracheo-innominate artery fistula is a highly lethal complication after tracheostomy. A 37-year-old man who had undergone a tracheostomy 14 years earlier because of dysphagia after brain surgery had a tracheo-innominate artery fistula with exsanguinating hemorrhage from his tracheostomy site. After temporary control of the bleeding, a stent graft was implanted in the innominate artery through the brachial artery. The patient recovered uneventfully and remained well 14 months after the procedure, with no sign of infection. Endovascular stent grafting may be the treatment of choice for patients with tracheo-innominate artery fistula. PMID- 11389431 TI - The problem of healing of endovascular stent grafts in the repair of abdominal aortic aneurysms. PMID- 11389432 TI - Regarding "fact and fiction surrounding the discovery of the venous valves". PMID- 11389434 TI - Regarding "aneurysm sac pressure measurements after endovascular repair of abdominal aortic aneurysms". PMID- 11389436 TI - Dynamics of the COPII coat with GTP and stable analogues. AB - We have developed an assay to monitor the assembly of the COPII coat onto liposomes in real time. We show that with Sar1pGTP bound to liposomes, a single round of assembly and disassembly of the COPII coat lasts a few seconds. The two large COPII complexes Sec23/24p and Sec13/31p bind almost instantaneously (in less than 1 s) to Sar1pGTP-doped liposomes. This binding is followed by a fast (less than 10 s) disassembly due to a 10-fold acceleration of the GTPase activating protein activity of Sec23/24p by the Sec13/31p complex. Experiments with the phosphate analogue BeFx suggest that Sec23/24p provides residues directly involved in GTP hydrolysis on Sar1p. PMID- 11389437 TI - Drosophila Pin1 prolyl isomerase Dodo is a MAP kinase signal responder during oogenesis. AB - The mammalian cis-trans prolyl isomerase Pin1 and its yeast orthologue Ess1/Ptf1 have been implicated in cell cycle control but a correlation between biochemical and physiological functions has not been established conclusively. Pin1 targets the proline residue carboxy-terminal to the phosphorylated threonine or serine residue, which constitutes part of the phosphorylated mitogen-activated protein kinase (MAPK) site PXpT/SP. Here we show that the Drosophila Pin1 homologue, the Dodo protein, is involved in dorsoventral patterning of the follicular epithelium in the egg chamber. Its function is to facilitate the degradation of transcription factor CF2, which requires, a priori, activated epidermal growth factor receptor-MAPK signalling. PMID- 11389438 TI - Actin dynamics at pointed ends regulates thin filament length in striated muscle. AB - Regulation of actin dynamics at filament ends determines the organization and turnover of actin cytoskeletal structures. In striated muscle, it is believed that tight capping of the fast-growing (barbed) ends by CapZ and of the slow growing (pointed) ends by tropomodulin (Tmod) stabilizes the uniform lengths of actin (thin) filaments in myofibrils. Here we demonstrate for the first time that both CapZ and Tmod are dynamic on the basis of the rapid incorporation of microinjected rhodamine-labelled actin (rho-actin) at both barbed and pointed ends and from the photobleaching of green fluorescent protein (GFP)-labelled Tmod. Unexpectedly, the inhibition of actin dynamics at pointed ends by GFP-Tmod overexpression results in shorter thin filaments, whereas the inhibition of actin dynamics at barbed ends by cytochalasin D has no effect on length. These data demonstrate that the actin filaments in myofibrils are relatively dynamic despite the presence of capping proteins, and that regulated actin assembly at pointed ends determines the length of thin filaments. PMID- 11389439 TI - Apaf-1 is a transcriptional target for E2F and p53. AB - Loss of function of the retinoblastoma protein, pRB, leads to lack of differentiation, hyperproliferation and apoptosis. Inactivation of pRB results in deregulated E2F activity, which in turn induces entry to S-phase and apoptosis. Induction of apoptosis by either the loss of pRB or the deregulation of E2F activity occurs via both p53-dependent and p53-independent mechanisms. The mechanism by which E2F induces apoptosis is still unclear. Here we show that E2F1 directly regulates the expression of Apaf-1, the gene for apoptosis protease activating factor 1. These results provide a direct link between the deregulation of the pRB pathway and apoptosis. Furthermore, because the pRB pathway is functionally inactivated in most cancers, the identification of Apaf-1 as a transcriptional target for E2F might explain the increased sensitivity of tumour cells to chemotherapy. We also show that, independently of the pRB pathway, Apaf 1 is a direct transcriptional target of p53, suggesting that p53 might sensitize cells to apoptosis by increasing Apaf-1 levels. PMID- 11389440 TI - FGF receptor signalling is required to maintain neural progenitors during Hensen's node progression. AB - Previous analyses of labelled clones of cells within the developing nervous system of the mouse have indicated that descendants are initially dispersed rostrocaudally followed by more local proliferation, which is consistent with the progressing node's contributing descendants from a resident population of progenitor cells as it advances caudally. Here we electroporated an expression vector encoding green fluorescent protein into the chicken embryo near Hensen's node to test and confirm the pattern inferred in the mouse. This provides a model in which a proliferative stem zone is maintained in the node by a localized signal; those cells that are displaced out of the stem zone go on to contribute to the growing axis. To test whether fibroblast growth factor (FGF) signalling could be involved in the maintenance of the stem zone, we co-electroporated a dominant-negative FGF receptor with a lineage marker, and found that it markedly alters the elongation of the spinal cord primordium. The results indicate that FGF receptor signalling promotes the continuous development of the posterior nervous system by maintaining presumptive neural progenitors in the region near Hensen's node. This offers a potential explanation for the mixed findings on FGF in the growth and patterning of the embryonic axis. PMID- 11389441 TI - Rme-1 regulates the distribution and function of the endocytic recycling compartment in mammalian cells. AB - RME-1 is an Eps15-homology (EH)-domain protein that was identified in a genetic screen for endocytosis genes in Caenorhabditis elegans. When expressed in a CHO cell line, the worm RME-1 protein and a mouse homologue are both associated with the endocytic recycling compartment. Here we show that expression of a dominant negative construct with a point mutation near the EH domain results in redistribution of the endocytic recycling compartment and slowing down of transferrin receptor recycling. The delivery of a TGN38 chimaeric protein to the trans-Golgi network is also slowed down. The function of Rme-1 in endocytic recycling is evolutionarily conserved in metazoans as shown by the protein's properties in C. elegans. PMID- 11389442 TI - Evidence that RME-1, a conserved C. elegans EH-domain protein, functions in endocytic recycling. AB - In genetic screens for new endocytosis genes in Caenorhabditis elegans we identified RME-1, a member of a conserved class of Eps15-homology (EH)-domain proteins. Here we show that RME-1 is associated with the periphery of endocytic organelles, which is consistent with a direct role in endocytic transport. Endocytic defects in rme-1 mutants indicate that the protein is likely to have a function in endocytic recycling. Evidence from studies of mammalian RME-1 also points to a function for RME-1 in recycling, specifically in the exit of membrane proteins from recycling endosomes. These studies show a conserved function in endocytic recycling for the RME-1 family of EH proteins. PMID- 11389443 TI - Regulation of c-myc expression by PDGF through Rho GTPases. AB - Src family protein-tyrosine kinases have a central role in several biological functions, including cell adhesion and spreading, chemotaxis, cell cycle progression, differentiation and apoptosis. Surprisingly, these kinases also participate in mitogenic signalling by receptors that themselves exhibit an intrinsic protein-tyrosine kinase activity, including those for platelet-derived growth factor (PDGF), epidermal growth factor and colony-stimulating factor-1. Indeed, Src kinases are strictly required for the nuclear expression of the c-myc proto-oncogene and thus for DNA synthesis in response to PDGF. However, the nature of the signalling pathways by which Src kinases participate in the induction of c-myc expression by tyrosine kinase receptors is still unknown. Here we show that PDGF enhances c-myc expression and stimulates the c-myc promoter in a Src-dependent manner, and that neither Ras nor the mitogen-activated protein kinase pathway mediate these effects. In contrast, we present evidence that PDGF stimulates Vav2 through Src, thereby initiating the activation of a Rac-dependent pathway that controls the expression of the c-myc proto-oncogene. PMID- 11389444 TI - TGF-beta induces assembly of a Smad2-Smurf2 ubiquitin ligase complex that targets SnoN for degradation. AB - The receptor-regulated Smad proteins are essential intracellular mediators of signal transduction by the transforming growth factor-beta (TGF-beta) superfamily of growth factors and are also important as regulators of gene transcription. Here we describe a new role for TGF-beta-regulated Smad2 and Smad3 as components of a ubiquitin ligase complex. We show that in the presence of TGF-beta signalling, Smad2 interacts through its proline-rich PPXY motif with the tryptophan-rich WW domains of Smurf2, a recently identified E3 ubiquitin ligases. TGF-beta also induces the association of Smurf2 with the transcriptional co repressor SnoN and we show that Smad2 can function to mediate this interaction. This allows Smurf2 HECT domain to target SnoN for ubiquitin-mediated degradation by the proteasome. Thus, stimulation by TGF-beta can induce the assembly of a Smad2-Smurf2 ubiquitin ligase complex that functions to target substrates for degradation. PMID- 11389445 TI - The translational inhibitor 4E-BP is an effector of PI(3)K/Akt signalling and cell growth in Drosophila. AB - The initiation factor 4E for eukaryotic translation (eIF4E) binds the messenger RNA 5'-cap structure and is important in the regulation of protein synthesis. Mammalian eIF4E activity is inhibited when the initiation factor binds to the translational repressors, the 4E-binding proteins (4E-BPS). Here we show that the Drosophila melanogaster 4E-BP (d4E-BP) is a downstream target of the phosphatidylinositol-3-OH kinase (PI(3)K) signal-transduction cascade, which affects the interaction of d4E-BP with eIF4E. Ectopic expression of a highly active d4E-BP mutant in wing-imaginal discs causes a reduction of wing size, brought about by a decrease in cell size and number. A marked reduction in cell size was also observed in post-mitotic cells. Expression of d4E-BP in the eye and wing together with PI(3)K or dAkt1, the serine/threonine kinase downstream of PI(3)K, resulted in suppression of the growth phenotype elicited by these kinases. Our results support a role for d4E-BP as an effector of cell growth. PMID- 11389446 TI - Expression of alpha- and beta-globin genes occurs within different nuclear domains in haemopoietic cells. AB - The alpha- and beta-globin gene clusters have been extensively studied. Regulation of these genes ensures that proteins derived from both loci are produced in balanced amounts, and that expression is tissue-restricted and specific to developmental stages. Here we compare the subnuclear location of the endogenous alpha- and beta-globin loci in primary human cells in which the genes are either actively expressed or silent. In erythroblasts, the alpha- and beta globin genes are localized in areas of the nucleus that are discrete from alpha satellite-rich constitutive heterochromatin. However, in cycling lymphocytes, which do not express globin genes, the distribution of alpha- and beta-globin genes was markedly different. beta-globin loci, in common with several inactive genes studied here (human c-fms and SOX-1) and previously (mouse lambda5, CD4, CD8alpha, RAGs, TdT and Sox-1), were associated with pericentric heterochromatin in a high proportion of cycling lymphocytes. In contrast, alpha-globin genes were not associated with centromeric heterochromatin in the nucleus of normal human lymphocytes, in lymphocytes from patients with alpha-thalassaemia lacking the regulatory HS-40 element or entire upstream region of the alpha-globin locus, or in mouse erythroblasts and lymphocytes derived from human alpha-globin transgenic mice. These data show that the normal regulated expression of alpha- and beta globin gene clusters occurs in different nuclear environments in primary haemopoietic cells. PMID- 11389447 TI - Direct, high-resolution measurement of furrow stiffening during division of adherent cells. AB - It is unclear whether cell division is driven by cortical relaxation outside the equatorial region or cortical contractility within the developing furrow alone. To approach this question, a technique is required that can monitor spatially resolved changes in cortical stiffness with good time resolution. We employed atomic force microscopy (AFM), in force-mapping mode, to track dynamic changes in the stiffness of the cortex of adherent cultured cells along a single scan-line during M phase, from metaphase to cytokinesis. Video microscopy, which we used to correlate the AFM data with mitotic events identified by light microscopy, indicated that the AFM force-mapping technique does not perturb dividing cells. Here we show that cortical stiffening occurs over the equatorial region about 160 seconds before any furrow appears, and that this stiffening markedly increases as the furrow starts. By contrast, polar relaxation of cells does not seem to be an obligatory event for cell division to occur. PMID- 11389448 TI - Making life a bit easier. PMID- 11389449 TI - Round-trip ticket: recycling to the plasma membrane requires RME-1. PMID- 11389450 TI - Passage through the nuclear pore. PMID- 11389451 TI - Pin-pointing MAPK signalling. PMID- 11389452 TI - Building from the bottom up. PMID- 11389453 TI - Membrane-cytoskeletal dynamics in a new dimension. AB - The recent Airlie House meeting on 'Cytoplasmic Organization and Membrane Traffic' (22-25 March 2001), sponsored by the Keith Porter Endowment, proved not to be the typical exchange of advances among specialists familiar with each other's work, but rather a series of interesting and diverse presentations that together illuminated the pace and pattern of membrane and cytoskeletal interactions in living cells. PMID- 11389456 TI - From fixed to FRAP: measuring protein mobility and activity in living cells. AB - Experiments with fluorescence recovery after photobleaching (FRAP) started 30 years ago to visualize the lateral mobility and dynamics of fluorescent proteins in living cells. Its popularity increased when non-invasive fluorescent tagging became possible with the green fluorescent protein (GFP). Many researchers use GFP to study the localization of fusion proteins in fixed or living cells, but the same fluorescent proteins can also be used to study protein mobility in living cells. Here we review the potential of FRAP to study protein dynamics and activity within a single living cell. These measurements can be made with most standard confocal laser-scanning microscopes equipped with photobleaching protocols. PMID- 11389457 TI - Bioinformatics beyond sequence: mapping gene function in the embryo. AB - The spatio-temporal expression pattern of a gene during development is a valuable piece of information. But there is no way to compare precisely the patterns of expression of different genes, or the way the patterns are changed in a mutant. One way to solve this problem is to construct digital reference images of development (a bioinformatics framework), to which expression patterns can be mapped and stored, then compared. Such frameworks are under active development in several model systems. They will form the basis of powerful and integrated gene expression databases, which facilitate comparisons between genes, tissues and species. PMID- 11389458 TI - Computational analysis of microarray data. AB - Microarray experiments are providing unprecedented quantities of genome-wide data on gene-expression patterns. Although this technique has been enthusiastically developed and applied in many biological contexts, the management and analysis of the millions of data points that result from these experiments has received less attention. Sophisticated computational tools are available, but the methods that are used to analyse the data can have a profound influence on the interpretation of the results. A basic understanding of these computational tools is therefore required for optimal experimental design and meaningful data analysis. PMID- 11389459 TI - Evolutionary computation. AB - Evolution does not require DNA, or even living organisms. In computer science, the field known as 'evolutionary computation' uses evolution as an algorithmic tool, implementing random variation, reproduction and selection by altering and moving data within a computer. This harnesses the power of evolution as an alternative to the more traditional ways to design software or hardware. Research into evolutionary computation should be of interest to geneticists, as evolved programs often reveal properties - such as robustness and non-expressed DNA - that are analogous to many biological phenomena. PMID- 11389460 TI - Finding the molecular basis of quantitative traits: successes and pitfalls. AB - Understanding the molecular basis of quantitative genetic variation is a principal goal for biomedicine. Although the complex genetic architecture of quantitative traits has so far largely frustrated attempts to identify genes in humans by standard linkage methodologies, quantitative trait loci (QTL) have been mapped in plants, insects and rodents. However, identifying the molecular bases of QTL remains a challenge. Here, we discuss why this is and how new experimental strategies and analytical techniques, combined with the fruits of the genome projects, are beginning to identify candidate genes for QTL studies in several model organisms. PMID- 11389461 TI - The emerging genetic and molecular basis of Fanconi anaemia. AB - The past few years have witnessed a considerable expansion in our understanding of the pathways that maintain chromosome stability in dividing cells through the identification of genes that are mutated in certain human chromosome instability disorders. Cells that are derived from patients with Fanconi anaemia (FA) show spontaneous chromosomal instability and mutagen hypersensitivity, but FA poses a unique challenge as the nature of the DNA-damage-response pathway thought to be affected by the disease has long been a mystery. However, the recent cloning of most of the FA-associated genes, and the characterization of their protein products, has provided tantalizing clues as to the molecular basis of this disease. PMID- 11389462 TI - Genetics of craniofacial development and malformation. AB - The head is anatomically the most sophisticated part of the body and its evolution was fundamental to the origin of vertebrates; understanding its development is a formidable problem in biology. A synthesis of embryology, evolution and mouse genetics is shaping our understanding of head development and in this review we discuss its application to studies of human craniofacial malformations. Many of these disorders have their origins in specific embryological processes, including abnormalities of brain patterning, of the migration and fusion of tissues in the face, and of bone differentiation in the skull vault. PMID- 11389463 TI - T. H. Morgan's resistance to the chromosome theory. PMID- 11389464 TI - The natural history of Caenorhabditis elegans research. PMID- 11389465 TI - Mechanisms of transcriptional memory. AB - How can the same gene remember that it is 'off' in one cell lineage and 'on' in another? Studies of how homeotic genes are regulated in Drosophila melanogaster have uncovered a transcriptional maintenance system, encoded by the Polycomb and trithorax group genes, that preserves expression patterns across development. Here we try to formulate a broad framework for the types of molecular mechanism used by the Polycomb and trithorax proteins. PMID- 11389466 TI - Protein modules that manipulate histone tails for chromatin regulation. AB - Histones are the predominant protein components of chromatin and are subject to specific post-translational modifications that are correlated with transcriptional competence. Among these histone modifications are acetylation, phosphorylation and methylation, and recent studies reveal that conserved protein modules mediate the attachment, removal or recognition of these modifications. It is becoming clear that appropriate coordination of histone modifications and their manipulations by conserved protein modules are integral to gene-specific transcriptional regulation within chromatin. PMID- 11389467 TI - The junction-resolving enzymes. AB - Junction-resolving enzymes are ubiquitous nucleases that are important for DNA repair and recombination and act on DNA molecules containing branch points, especially four-way junctions. They show a pronounced selectivity for the structure of the DNA substrate but, despite its importance, the structural selectivity is not well understood. This poses an intriguing challenge in molecular recognition on a relatively large scale. PMID- 11389468 TI - Studying protein dynamics in living cells. AB - Since the advent of the green fluorescent protein, the subcellular localization, mobility, transport routes and binding interactions of proteins can be studied in living cells. Live cell imaging, in combination with photobleaching, energy transfer or fluorescence correlation spectroscopy are providing unprecedented insights into the movement of proteins and their interactions with cellular components. Remarkably, these powerful techniques are accessible to non specialists using commercially available microscope systems. PMID- 11389469 TI - Living dangerously: how Helicobacter pylori survives in the human stomach. AB - Helicobacter pylori was already present in the stomach of primitive humans as they left Africa and spread through the world. Today, it still chronically infects more than 50% of the human population, causing, in some cases, severe diseases such as peptic ulcers and stomach cancer. To succeed in these long-term associations, H. pylori has developed a unique set of virulence factors, which allow survival in a unique and hostile ecological niche--the human stomach. PMID- 11389470 TI - The hunting of the Src. AB - The non-receptor tyrosine kinase Src is important for many aspects of cell physiology. The viral src gene was the first retroviral oncogene to be identified, and its cellular counterpart was the first proto-oncogene to be discovered in the vertebrate genome. Src has been important, not only as an object of study in itself, but also as an entry point into the molecular genetics of cancer. PMID- 11389471 TI - From Galen to Golgi: birth of the life sciences in Italy. AB - Between the sixteenth and twentieth centuries, the study of biology was intimately intertwined with progress in medicine. So how, when and where did research into the life sciences begin? PMID- 11389472 TI - The TRP ion channel family. AB - Mammalian homologues of the Drosophila transient receptor potential (TRP) channel gene encode a family of at least 20 ion channel proteins. They are widely distributed in mammalian tissues, but their specific physiological functions are largely unknown. A common theme that links the TRP channels is their activation or modulation by phosphatidylinositol signal transduction pathways. The channel subunits have six transmembrane domains that most probably assemble into tetramers to form non-selective cationic channels, which allow for the influx of calcium ions into cells. Three subgroups comprise the TRP channel family; the best understood of these mediates responses to painful stimuli. Other proposed functions include repletion of intracellular calcium stores, receptor-mediated excitation and modulation of the cell cycle. PMID- 11389473 TI - Neuromodulation of Na+ channels: an unexpected form of cellular plasticity. AB - Voltage-gated Na+ channels set the threshold for action potential generation and are therefore good candidates to mediate forms of plasticity that affect the entire neuronal output. Although early studies led to the idea that Na+ channels were not subject to modulation, we now know that Na+ channel function is affected by phosphorylation. Furthermore, Na+ channel modulation is implicated in the control of input-output relationships in several types of neuron and seems to be involved in phenomena as varied as cocaine withdrawal, hyperalgesia and light adaptation. Here we review the available evidence for the regulation of Na+ channels by phosphorylation, its molecular mechanism, and the possible ways in which it affects neuronal function. PMID- 11389474 TI - Protein-protein interactions, cytoskeletal regulation and neuronal migration. AB - Neuronal migration, like the migration of many cell types, requires an extensive rearrangement of cell shape, mediated by changes in the cytoskeleton. The genetic analysis of human brain malformations has identified several biochemical players in this process, including doublecortin (DCX) and LIS1, mutations of which cause a profound migratory disturbance known as lissencephaly ('smooth brain') in humans. Studies in mice have identified additional molecules such as Cdk5, P35, Reelin, Disabled and members of the LDL superfamily of receptors. Understanding the cell biology of these molecules has been a challenge, and it is not known whether they function in related biochemical pathways or in very distinct processes. The last year has seen rapid advances in the biochemical analysis of several such molecules. This analysis has revealed roles for some of these molecules in cytoskeletal regulation and has shown an unexpected conservation of the genetic pathways that regulate neuronal migration in humans and nuclear movement in simple eukaryotic organisms. PMID- 11389475 TI - Primate anterior cingulate cortex: where motor control, drive and cognition interface. AB - Controversy surrounds the function of the anterior cingulate cortex. Recent discussions about its role in behavioural control have centred on three main issues: its involvement in motor control, its proposed role in cognition and its relationship with the arousal/drive state of the organism. I argue that the overlap of these three domains is key to distinguishing the anterior cingulate cortex from other frontal regions, placing it in a unique position to translate intentions to actions. PMID- 11389476 TI - Electrical synapses between GABA-releasing interneurons. AB - Although gap junctions were first demonstrated in the mammalian brain about 30 years ago, the distribution and role of electrical synapses have remained elusive. A series of recent reports has demonstrated that inhibitory interneurons in the cerebral cortex, thalamus, striatum and cerebellum are extensively interconnected by electrical synapses. Investigators have used paired recordings to reveal directly the presence of electrical synapses among identified cell types. These studies indicate that electrical coupling is a fundamental feature of local inhibitory circuits and suggest that electrical synapses define functionally diverse networks of GABA-releasing interneurons. Here, we discuss these results, their possible functional significance and the insights into neuronal circuit organization that have emerged from them. PMID- 11389477 TI - Sharing gene expression data: an array of options. AB - Sharing of microarray data has many advantages for the scientific and biomedical community, and should be advocated by neuroscience journals. The goals of sharing are manifold, and include improving analysis and confidence in results, and facilitating global comparisons between experiments, while at the same time, not penalizing those who share. The sharing of microarray data poses unique challenges relative to more generic data such as DNA sequences. These challenges are surmountable, and various sharing formats are possible. Centralized non commercial databases are being developed to facilitate this process. PMID- 11389478 TI - The sharing of cDNA microarray data. AB - During the initial development of microarrays, much discussion revolved around the technology itself. The discussion has now shifted to data analysis and data sharing. There is great interest in the sharing of cDNA microarray data, but several issues related to format, quality and validation will need to be resolved before microarray data can be meaningfully integrated into other molecular databases. PMID- 11389479 TI - Microarrays: lost in a storm of data? AB - Microarray expression profiling is instrumental to our understanding of the function of the genome. Resolution of functionally relevant expression patterns will require the analysis of large data sets compiled from multiple investigators. For this and other reasons, I argue that it is crucial for array data to be publicly shared in a format as close to the 'raw data' as possible. Issues such as protection of intellectual property, ensuring quality of the data, and the format and timing for sharing array data are also discussed. PMID- 11389480 TI - Microarrays in brain research: the good, the bad and the ugly. AB - Making sense of microarray data is a complex process, in which the interpretation of findings will depend on the overall experimental design and judgement of the investigator performing the analysis. As a result, differences in tissue harvesting, microarray types, sample labelling and data analysis procedures make post hoc sharing of microarray data a great challenge. To ensure rapid and meaningful data exchange, we need to create some order out of the existing chaos. In these ground-breaking microarray standardization and data sharing efforts, NIH agencies should take a leading role PMID- 11389481 TI - A schizophrenia-susceptibility locus at 6q25, in one of the world's largest reported pedigrees. AB - We have completed a genome scan of a 12-generation, 3,400-member pedigree with schizophrenia. Samples from 210 individuals were collected from the pedigree. We performed an "affecteds-only" genome-scan analysis using 43 members of the pedigree. The affected individuals included 29 patients with schizophrenia, 10 with schizoaffective disorders, and 4 with psychosis not otherwise specified. Two sets of white-European allele frequencies were used-one from a Swedish control population (46 unrelated individuals) and one from the pedigree (210 individuals). All analyses pointed to the same region: D6S264, located at 6q25.2, showed a maximum LOD score of 3.45 when allele frequencies in the Swedish control population were used, compared with a maximum LOD score of 2.59 when the pedigree's allele frequencies were used. We analyzed additional markers in the 6q25 region and found a maximum LOD score of 6.6 with marker D6S253, as well as a 6-cM haplotype (markers D6S253-D6S264) that segregated, after 12 generations, with the majority of the affected individuals. Multipoint analysis was performed with the markers in the 6q25 region, and a maximum LOD score of 7.7 was obtained. To evaluate the significance of the genome scan, we simulated the complete analysis under the assumption of no linkage. The results showed that a LOD score >2.2 should be considered as suggestive of linkage, whereas a LOD score >3.7 should be considered as significant. These results suggest that a common ancestral region was inherited by the affected individuals in this large pedigree. PMID- 11389482 TI - Single-amino-acid deletion in the RYR1 gene, associated with malignant hyperthermia susceptibility and unusual contraction phenotype. AB - Malignant hyperthermia (MH) is an anesthetic-drug-induced, life-threatening hypermetabolic syndrome caused by abnormal calcium regulation in skeletal muscle. Often inherited as an autosomal dominant trait, MH has linkage to 30 different mutations in the RYR1 gene, which encodes a calcium-release-channel protein found in the sarcoplasmic reticulum membrane in skeletal muscle. All published RYR1 mutations exclusively represent single-nucleotide changes. The present report documents, in exon 44 of RYR1 in two unrelated, MH-susceptible families, a 3-bp deletion that results in deletion of a conserved glutamic acid at position 2347. This is the first deletion, in RYR1, found to be associated with MH susceptibility. MH susceptibility was confirmed among some family members by in vitro diagnostic pharmacological contracture testing of biopsied skeletal muscle. Although a single-amino-acid deletion appears to be a subtle change in the protein, the deletion of Glu2347 from RYR1 produces an unusually large electrically evoked contraction tension in MH-positive individuals, suggesting that this deletion produces an alteration in skeletal-muscle calcium regulation, even in the absence of pharmacological agents. PMID- 11389483 TI - Leber congenital amaurosis and retinitis pigmentosa with Coats-like exudative vasculopathy are associated with mutations in the crumbs homologue 1 (CRB1) gene. AB - Mutations in the crumbs homologue 1 (CRB1) gene cause a specific form of retinitis pigmentosa (RP) that is designated "RP12" and is characterized by a preserved para-arteriolar retinal pigment epithelium (PPRPE) and by severe loss of vision at age <20 years. Because of the early onset of disease in patients who have RP with PPRPE, we considered CRB1 to be a good candidate gene for Leber congenital amaurosis (LCA). Mutations were detected in 7 (13%) of 52 patients with LCA from the Netherlands, Germany, and the United States. In addition, CRB1 mutations were detected in five of nine patients who had RP with Coats-like exudative vasculopathy, a relatively rare complication of RP that may progress to partial or total retinal detachment. Given that four of five patients had developed the complication in one eye and that not all siblings with RP have the complication, CRB1 mutations should be considered an important risk factor for the Coats-like reaction, although its development may require additional genetic or environmental factors. Although no clear-cut genotype-phenotype correlation could be established, patients with LCA, which is the most severe retinal dystrophy, carry null alleles more frequently than do patients with RP. Our findings suggest that CRB1 mutations are a frequent cause of LCA and are strongly associated with the development of Coats-like exudative vasculopathy in patients with RP. PMID- 11389484 TI - The Silver syndrome variant of hereditary spastic paraplegia maps to chromosome 11q12-q14, with evidence for genetic heterogeneity within this subtype. AB - The hereditary spastic paraplegias (HSPs) are a complex group of neurodegenerative disorders characterized by lower-limb spasticity and weakness. Silver syndrome (SS) is a particularly disabling dominantly inherited form of HSP, complicated by amyotrophy of the hand muscles. Having excluded the multiple known HSP loci, we undertook a genomewide screen for linkage of SS in one large multigenerational family, which revealed evidence for linkage of the SS locus, which we have designated "SPG17," to chromosome 11q12-q14. Haplotype construction and analysis of recombination events permitted the minimal interval defining SPG17 to be refined to approximately 13 cM, flanked by markers D11S1765 and D11S4136. SS in a second family was not linked to SPG17, demonstrating further genetic heterogeneity in HSP, even within this clinically distinct subtype. PMID- 11389485 TI - Disorders of peroxisome biogenesis due to mutations in PEX1: phenotypes and PEX1 protein levels. AB - Zellweger syndrome (ZS), neonatal adrenoleukodystrophy (NALD), and infantile Refsum disease (IRD) are clinically overlapping syndromes, collectively called "peroxisome biogenesis disorders" (PBDs), with clinical features being most severe in ZS and least pronounced in IRD. Inheritance of these disorders is autosomal recessive. The peroxisome biogenesis disorders are genetically heterogeneous, having at least 12 different complementation groups (CGs). The gene affected in CG1 is PEX1. Approximately 65% of the patients with PBD harbor mutations in PEX1. In the present study, we used SSCP analysis to evaluate a series of patients belonging to CG1 for mutations in PEX1 and studied phenotype genotype correlations. A complete lack of PEX1 protein was found to be associated with severe ZS; however, residual amounts of PEX1 protein were found in patients with the milder phenotypes, NALD and IRD. The majority of these latter patients carried at least one copy of the common G843D allele. When patient fibroblasts harboring this allele were grown at 30 degrees C, a two- to threefold increase in PEX1 protein levels was observed, associated with a recovery of peroxisomal function. This suggests that the G843D missense mutation results in a misfolded protein, which is more stable at lower temperatures. We conclude that the search for the factors and/or mechanisms that determine the stability of mutant PEX1 protein by high-throughput procedures will be a first step in the development of therapeutic strategies for patients with mild PBDs. PMID- 11389487 TI - Imported Fasciola hepatica infection in the United States and treatment with triclabendazole. AB - Infection with Fasciola hepatica, a liver trematode, is not frequently reported in the United States. We describe 2 patients, both originally from Cape Verde, who illustrate the spectrum of clinical presentations of F. hepatica as well as the means of treating infection with this parasite. Patient 1 had extensive disease and underwent multiple diagnostic procedures before the correct diagnosis was reached. Patient 2, who had few symptoms, had fascioliasis diagnosed by a noninvasive evaluation. Both patients were treated with triclabendazole without experiencing significant side effects. Fascioliasis that has been imported to the United States may elude prompt or accurate diagnosis. Obtaining a detailed travel history and recognizing the clinical presentation early in the course of infection may permit timely and noninvasive identification of infection. Triclabendazole is now the recommended drug for treating for fascioliasis because of its efficacy, safety, and ease of use. PMID- 11389486 TI - A mutation, in the iron-responsive element of H ferritin mRNA, causing autosomal dominant iron overload. AB - Ferritin, which is composed of H and L subunits, plays an important role in iron storage and in the control of intracellular iron distribution. Synthesis of both ferritin subunits is controlled by a common cytosolic protein, iron regulatory protein (IRP), which binds to the iron-responsive element (IRE) in the 5'-UTR of the H- and L-ferritin mRNAs. In the present study, we have identified a single point mutation (A49U) in the IRE motif of H-ferritin mRNA, in four of seven members of a Japanese family affected by dominantly inherited iron overload. Gel shift mobility assay and Scatchard-plot analysis revealed that a mutated IRE probe had a higher binding affinity to IRP than did the wild-type probe. When mutated H subunit was overexpressed in COS-1 cells, suppression of H-subunit synthesis and of the increment of radiolabeled iron uptake were observed. These data suggest that the A49U mutation in the IRE of H-subunit is responsible for tissue iron deposition and is a novel cause of hereditary iron overload, most likely related to impairment of the ferroxidase activity generated by H subunit. PMID- 11389488 TI - Necrotizing fasciitis associated with injection drug use. AB - We studied cases of necrotizing fasciitis among injection drug users (IDUs) and non-IDUs who presented at the University of California Davis Medical Center from 1984 through 1999. Of 107 patients, 59 (55%) were IDUs and 48 (45%) non-IDUs. Among IDUs, 32 (54%) recently injected at the site of infection, and 17 patients (29%) presented with an abscess. Among non-IDUs, 17 (35%) reported a recent insect bite and 9 (19%) reported a wound or abrasion at the site of infection. Overall, seventy cases (65%) had > or = 3 debridements, and 31 patients (29%) had > 5% of their total body surface area debrided. Of all patients with necrotizing fasciitis, 16 (15%) did not survive. Among the 59 IDUs, 6 (10%) did not survive, while among non-IDUs, 10 (21%) did not survive. Our results indicate the need for a high index of suspicion for necrotizing fasciitis among patients presenting with cellulitis, a recent insect bite, wound, or recent injection drug use. Preventive interventions for necrotizing fasciitis among IDUs should include street-based education and treatment for abscesses and cellulitis. PMID- 11389489 TI - Pneumococcal infections in children after transplantation. AB - Bacterial infections in recipients of bone marrow and solid-organ transplants remain a major cause of morbidity and death. The cases of 42 children who had undergone transplantation and developed an infection with Streptococcus pneumoniae were retrospectively reviewed. Thirty-four patients had 1 episode of infection, whereas 7 had 2 episodes and 1 had 3 episodes of infection. Solid organ recipients were more likely to have recurrent invasive disease (P<.02). A total of 31 (74%) of 42 patients were on immunosuppressive therapy, and 74% had been on antimicrobial therapy within 30 days before diagnosis of S. pneumoniae infection. Only 33% of eligible patients had received a pneumococcal vaccine. Twenty-six percent of isolates recovered were not susceptible to penicillin, and 18% were not susceptible to ceftriaxone. Two patients experienced infection related deaths; one of these had a penicillin-nonsusceptible isolate. The antimicrobial susceptibilities and outcome of infections with S. pneumoniae in patients who have undergone transplantation are similar to those in the general pediatric population. PMID- 11389490 TI - Treatment of patients with refractory giardiasis. AB - Giardia lamblia is one of the most common parasitic infections. Although standard treatments are usually curative, some immunocompromised patients, including patients with acquired immunodeficiency syndrome as well as healthy patients, have giardiasis that is refractory to recommended regimens. We report our experience with 6 patients with giardiasis, for whom therapy with a combination of quinacrine and metronidazole resulted in cures for 5 of the 6 patients. PMID- 11389491 TI - Randomized, double-blind, controlled study of cefoperazone-sulbactam plus cotrimoxazole versus ceftazidime plus cotrimoxazole for the treatment of severe melioidosis. AB - We conducted a prospective randomized, double-blind, controlled study of cefoperazone-sulbactam (ratio, 1:1; cefoperazone 25 mg/kg/day) plus cotrimoxazole (trimethoprim-sulfamethoxazole [TMP-SMZ] at a ratio of 80:400; TMP, 8 mg/kg/day) versus ceftazidime (100 mg/kg/day) plus cotrimoxazole (TMP, 8 mg/kg/day) for the treatment of severe melioidosis. Of 219 patients enrolled in the study, 102 (47%) had culture-proven melioidosis. These patients were assigned randomly to 2 treatment groups, each with 50 patients (2 patients were excluded). Mortality rates were not significantly different between the 2 groups: 18% in the cefoperazone-sulbactam group versus 14% in the ceftazidime group. The crude difference in the mortality rate was 4%, but when adjusted for type of infection the difference was 0.9% (95% confidence interval, -3.6% to 5.4%; P = .696). The duration of defervescence and the bacteriological response of successfully treated patients were similar in both groups, and both treatment regimens were well tolerated. Cefoperazone-sulbactam plus cotrimoxazole might be used as an alternative to ceftazidime plus cotrimoxazole as treatment for severe melioidosis. PMID- 11389492 TI - Risk factors for skin and soft-tissue abscesses among injection drug users: a case-control study. AB - Skin and soft-tissue abscesses, a common problem among injection drug users (IDUs), result in serious morbidity for the patient and costly hospitalizations for incision and drainage; however, there has been little etiologic or preventive epidemiologic research on this problem. We performed a case-control study that enrolled 151 IDUs who had been given a new diagnosis of abscess requiring incision and drainage (cases) and 267 IDUs who did not have abscess or other bacterial infection during the previous year and who were stratum-matched to cases according to age, sex, and race (controls). Subcutaneous or intramuscular, instead of intravenous, injection is a major risk factor for abscess among IDUs. The injection of a cocaine and heroin mixture, or "speedball," may predispose patients to develop abscess by inducing soft-tissue ischemia. Cleaning the skin with alcohol before injection was found to have a protective effect. Neither human immunodeficiency virus nor human T-lymphotropic virus type II seropositivity was significantly associated with abscess. PMID- 11389493 TI - Longitudinal study of bacterial, viral, and fungal infections in adult recipients of bone marrow transplants. AB - The epidemiology of infections was studied in a retrospective cohort of 446 recipients of bone marrow transplants (BMTs; 92 of which were allogeneic and 354 of which were autologous) during 1993--1996. Infections that were microbiologically documented in 274 recipients included bacteremia, urinary tract infections, cytomegalovirus viremia, fungemia, invasive aspergillosis, and catheter-related infections. During the period of neutropenia, no differences were found between recipients of allogeneic BMTs and recipients of autologous BMTs with regard to the incidence and the nature of infection. After patients underwent engraftment, bacteremia, cytomegalovirus viremia, and invasive aspergillosis were significantly more common in recipients of allogeneic BMTs than in recipients of autologous BMTs. Deaths caused by infection were uncommon and were mainly the result of invasive aspergillosis. Therefore, empirical antimicrobial therapy should be the same for recipients of both allogeneic and autologous BMTs during the period of neutropenia; after engraftment, more attention should be paid to the risk of infection in allogeneic BMT recipients, particularly with regard to detection and prevention of invasive aspergillosis. PMID- 11389494 TI - Bacteremia due to quinolone-resistant Escherichia coli in a teaching hospital in South Korea. AB - Quinolone-resistant Escherichia coli (QREC) strains are being isolated with increasing frequency. From 1993 to 1998, 40 cases of QREC bacteremia were observed in a teaching hospital; 25 episodes (63.5%) were community-acquired. The incidence of QREC bacteremia increased steadily, from 6.7% to 24.6% during 5 years, and correlated with the significantly increased use of fluoroquinolones (P = .003, r = 0.98). When the 40 QREC bacteremic patients were compared with 80 patients with bacteremia due to quinolone-susceptible E. coli, prior fluoroquinolone use was the only independent risk factor for QREC bacteremia (P = .001). A high APACHE II score was the only independent risk factor for death. The rate of multidrug resistance of QREC was much higher (60%) than that of quinolone susceptible isolates (13.8%). Pulsed-field gel electrophoresis patterns of these isolates were diverse. Therefore, the isolates revealed little evidence of clonal spread and may have emerged in direct response to the selective pressure exerted by prior fluoroquinolone use. PMID- 11389495 TI - Eikenella corrodens infections in children and adolescents: case reports and review of the literature. AB - Eikenella corrodens is a slow-growing, gram-negative, nonmotile, facultative rod that can cause infection in humans. Although the clinical characteristics of Eikenella infections in adults are well described, the literature regarding Eikenella infections in children is lacking. Thirteen cases of Eikenella infection in children and adolescents reported from a hospital and an additional 41 cases from the literature were reviewed. Eikenella species can be serious pediatric pathogens, particularly when there is an exposure to human oral secretions. Empirical therapy used to manage most oropharyngeal flora may be ineffective against Eikenella species. The treatment of choice for children and adolescents who are infected with Eikenella species includes a combination of surgical management and antibiotics. PMID- 11389496 TI - Characteristics of patients with herpes zoster on presentation to practitioners in France. AB - There have been many epidemiological studies of chickenpox but only a few of herpes zoster. We report data from an observational study, conducted in France during a 1-year period, of 9038 patients who presented with acute herpes zoster (n = 8103) or postherpetic neuralgia (PHN; n = 935) at the office practices of 4635 general practitioners or dermatologists. The incidence of herpes zoster in France was found to be similar to that in the literature: from 1.4 to 4.8 cases per 1000 population per year. The patient profiles and clinical patterns were delineated, as well as the management decisions made according to the type of treating physician. The impact of herpes zoster on quality of life was evaluated on the basis of the Medical Outcome Study Short Form 36 (MOS SF 36) scale, which is widely used for assessing quality of life in the field of health. This study provides reference data on the substantial deterioration in quality of life associated with herpes zoster and PHN. PMID- 11389497 TI - Hepatitis C virus genotypes and risk of cirrhosis in southern Italy. AB - Because hepatitis C virus (HCV) genotypes have raised considerable interest as variables that influence chronic hepatitis C progression, a case-control study was conducted to estimate their effects on patients with cirrhosis. Case patients (n = 46) had tested positive for anti-HCV antibody and HCV RNA and were residents of the study area who had cirrhosis recently diagnosed. Controls (n = 138) were drawn randomly from a residents' cohort from the same area. Demographic and other information were recorded. Presence of HCV infection, presence of HCV RNA, and HCV genotypes were assessed. Crude, stratified, and logistic regression analyses were performed. HCV genotype 2a/c occurred in 84 controls (60.9%) and 9 case patients (19.6%); HCV genotype 1b was found in 45 controls (32.6%) and 34 case patients (73.9%). HCV 1b genotype showed an independent effect on the risk of cirrhosis (odds ratio, 7.49; 95% confidence interval, 3.15--17.81). No significant effects related to other variables were observed. These results indicate that the genetic diversity of HCV phylogenetic variants may explain differences in biological behaviors. PMID- 11389498 TI - Pathways for inappropriate dispensing of antibiotics for rhinosinusitis: a randomized trial. AB - We evaluated the extent of and factors that determine the inappropriate use of antibiotics that are obtained without a physician's prescription. Ninety-eight Greek pharmacists were visited by actress-researchers who played clients requesting antibiotics without a physician's prescription. Pharmacists were randomly challenged in a scenario that involved simulated cases of acute uncomplicated rhinosinusitis with either low fever (38.5 degrees C) or high fever (40 degrees C). Antibiotics were offered by 34 (69%) of 49 pharmacists who were presented with the high-fever scenario and by 42 (86%) of 49 pharmacists who were presented with the low-fever scenario (risk difference, 16.3%; P = .05). Thirty two (65%) and 35 (71%) pharmacists in the high- and low-fever study arms, respectively, agreed to sell the actress-researchers broad-spectrum antibiotics. Only 28 (57%) and 17 (35%) pharmacists, respectively, recommended that the patient visit a physician (P = .03). Inappropriate recommendations regarding antibiotic use were very common in the studied setting. Antibiotics were more likely to be offered to persons who did not have a prescription when they were less likely to be clinically indicated. PMID- 11389499 TI - Diagnosis of atypical cases of infectious mononucleosis. AB - The variable manifestations of infectious mononucleosis rarely cause clinicians to suspect primary Epstein-Barr virus or cytomegalovirus infection; consequently, costly diagnostic tests and unnecessary treatments are undertaken. Seventeen cases of clinically atypical and 11 cases of clinically typical infectious mononucleosis were diagnosed through screening for atypical and apoptotic lymphocytes in the peripheral blood samples by means of an automated hematologic analyzer. Atypical and typical cases did not differ significantly with respect to peripheral white blood cell counts; percentages of lymphocytes, atypical lymphocytes, CD4(+) lymphocytes, human leukocyte antigen--DR positivity in CD3 lymphocytes, or apoptotic cells in blood smear after incubation; or levels of aspartate aminotransferase, alanine aminotransferase, and lactate dehydrogenase. Only the percentage of CD8(+) lymphocytes was significantly higher in patients with typical infectious mononucleosis than it was in patients with atypical infectious mononucleosis. Because certain atypical cases of infectious mononucleosis display laboratory abnormalities that are characteristic of typical infectious mononucleosis, enhanced awareness can help in the diagnosis. PMID- 11389500 TI - Antimicrobial resistance among uropathogens that cause community-acquired urinary tract infections in women: a nationwide analysis. AB - Current recommendations for empirical therapy for community-acquired urinary tract infection (UTI) in women hinge on knowledge of antimicrobial susceptibility patterns in the geographic region of the practitioner. We conducted a survey of antimicrobial susceptibilities of 103,223 isolates recovered from urine samples that were obtained in 1998 from female outpatients nationally and within 9 geographic regions in the United States. Resistance of Escherichia coli isolates to trimethoprim-sulfamethoxazole varied significantly according to geographic region, ranging from a high of 22% in the western United States to a low of 10% in the Northeast (P<.001). There were no clinically significant age-related differences in the susceptibility of E. coli to any of the study drugs, but the susceptibility to fluoroquinolones of non-E. coli isolates that were recovered from women who were aged >50 years was significantly lower than that of isolates recovered from younger women (P<.001). The in vitro susceptibility of uropathogens in female outpatients varies according to age and geographic region. PMID- 11389501 TI - Need for alternative trial designs and evaluation strategies for therapeutic studies of invasive mycoses. AB - Studies of invasive fungal infections have been and remain difficult to implement. Randomized clinical trials of fungal infections are especially slow and expensive to perform because it is difficult to identify eligible patients in a timely fashion, to prove the presence of the fungal infection in an unequivocal fashion, and to evaluate outcome in a convincing fashion. Because of these challenges, licensing decisions for antifungal agents have to date depended heavily on historical control comparisons and secondary advantages of the new agent. Although the availability of newer and potentially more effective agents makes these approaches less desirable, the fundamental difficulties of trials of invasive fungal infections have not changed. Therefore, there is a need for alternative trial designs and evaluation strategies for therapeutic studies of invasive mycoses, and this article summarizes the possible strategies in this area. PMID- 11389502 TI - Counterpoint: alternative trial designs for antifungal drugs--time to talk. PMID- 11389503 TI - Traveler's diarrhea due to intestinal protozoa. AB - Intestinal protozoa account for a minority of cases of acute traveler's diarrhea, but they are common pathogens in travelers who experience protracted diarrhea during or after travel. Evaluation of the traveler with chronic diarrhea should include a careful examination for typical infecting organisms, such as Giardia and Entamoeba species, as well as for emerging parasites, such as Cryptosporidium species, Cyclospora species, and microsporidia. The microbiology, epidemiology, clinical presentation, and treatment of the most common intestinal parasites found in travelers are presented in this minireview. PMID- 11389504 TI - Renal diseases associated with human immunodeficiency virus infection: epidemiology, clinical course, and management. AB - Human immunodeficiency virus (HIV)--associated nephropathy (HIVAN) and other glomerular lesions (e.g., immunoglobulin A nephropathy and immune complex glomerulonephritis) are frequent complications of HIV infection. These renal diseases usually present as a nephrotic syndrome with progressive loss of renal function and an increased risk of mortality. The prevalence and epidemiology of these renal lesions remain largely undefined; however, most studies agree that black race is a major risk factor for HIVAN. Observational studies have suggested that antiretroviral medications and angiotensin-converting enzyme inhibitors have beneficial effects on slowing the progression of renal disease among patients with HIVAN; however, little is known about the effect of these therapies on other renal lesions. Future research should focus on gaining a better understanding of the distribution and determinants of renal disease among HIV-infected patients as well as on performing controlled studies to test treatment strategies. PMID- 11389505 TI - Short-course therapy for right-side endocarditis due to Staphylococcus aureus in drug abusers: cloxacillin versus glycopeptides in combination with gentamicin. AB - A prospective, randomized clinical trial among drug abusers was conducted to assess the efficacy and safety of a short course of a combination of a glycopeptide (vancomycin or teicoplanin) and gentamicin compared with a combination of cloxacillin and gentamicin for treatment of right-side endocarditis caused by Staphylococcus aureus. Therapeutic success was significantly more frequent with cloxacillin than with a glycopeptide. No adverse effects were noted among patients in the cloxacillin group. A 14-day course of vancomycin or teicoplanin plus gentamicin is ineffective in this instance because it is associated with a high rate of clinical and microbiological failure. PMID- 11389506 TI - Control of an outbreak of infection due to extended-spectrum beta-lactamase- producing Escherichia coli in a liver transplantation unit. AB - We report an outbreak of infection due to genotypically identical extended spectrum beta-lactamase--producing Escherichia coli among patients in a liver transplantation unit. Control of the outbreak was achieved by a combination of contact isolation, feedback on hand hygiene, and gut decontamination with an orally administered fluoroquinolone. These interventions led to abrupt curtailment of the outbreak. PMID- 11389507 TI - Severe Bordetella holmesii infection in a previously healthy adolescent confirmed by gene sequence analysis. AB - We describe an immunocompetent adolescent who presented with exceptionally severe Bordetella holmesii infection, including previously undescribed manifestations. Sequelae included a severe restrictive lung defect due to pulmonary fibrosis. PMID- 11389508 TI - Mastitis due to Mycobacterium abscessus after body piercing. AB - We describe a patient with granulomatous mastitis due to Mycobacterium abscessus that presented as a mass lesion and was associated with a pierced nipple. To our knowledge, this is the first reported case of mastitis due to M. abscessus and the first association of this organism with body piercing. PMID- 11389509 TI - Staphylococcus epidermidis with intermediate resistance to vancomycin: elusive phenotype or laboratory artifact? AB - The recent and troubling isolation of Staphylococcus aureus and coagulase negative staphylococci that have increased resistance to glycopeptide antibiotics has prompted the use of aggressive surveillance measures in the clinical microbiology laboratory to aid in the recognition of these strains. Despite increasing awareness, the confirmation of glycopeptide resistance among staphylococci can be problematic; we present a case of catheter-associated peritonitis caused by Staphylococcus epidermidis to illustrate the dilemma. PMID- 11389510 TI - Subacute clinical forms of Plasmodium falciparum malaria in travelers receiving chloroquine-proguanil prophylaxis. AB - We have observed 4 French travelers, returning from African countries, who were not immune to malaria and were receiving chloroquine-proguanil prophylaxis, in whom the diagnosis of malaria could easily have been missed because the clinical signs were uncommon. These cases suggest that chloroquine-proguanil prophylaxis is not always effective and that travelers with unexplained symptoms should be monitored closely for malaria. PMID- 11389511 TI - Prevalence of the T215Y mutation in human immunodeficiency virus type 1-infected pregnant women in a New York cohort, 1995--1999. AB - From 1997 through 1999, the prevalence of the zidovudine resistance mutation T215Y was 9.7% among pregnant women, and the human immunodeficiency virus type 1 (HIV-1) load in those with resistant virus was higher than that measured in women with wild-type HIV-1. All mutations were noted in women with zidovudine experience, which suggests that monotherapy may not be adequate prophylaxis for vertical transmission of HIV-1 infection in the current era. PMID- 11389512 TI - "Helicobacter rappini" isolates from 2 homosexual men. AB - We report 2 cases of bacteremia due to "Helicobacter rappini" in 2 young, homosexual men, including the first report of H. rappini in a human immunodeficiency virus-positive patient. Blood cultures showed a spiral, fusiform, gram-negative bacterium with bipolar sheathed flagella. PMID- 11389513 TI - Recovery of anaerobic bacteria from 3 patients with infection at a pierced body site. AB - We describe 3 adolescents who developed infections due to anaerobes at pierced body sites: the nipple, the umbilicus, and the nasal septum. Anaerobes (Prevotella intermedia and Peptostreptococcus anaerobius) were recovered from pure culture of specimens obtained from 1 patient with nipple infection and were mixed with aerobic bacteria in cultures of specimens obtained from 2 patients (Streptococcus aureus, Peptostreptococcus micros, and Prevotella melaninogenica were recovered from a patient with nasal septum infection, and Bacteroides fragilis and Enterococcus faecalis were recovered from a patient with umbilical infection). The infection resolved in all patients after removal of the ornaments and use of antimicrobial drug treatment. PMID- 11389514 TI - Introduction to the Immunocompromised Host Society consensus conference on epidemiology, prevention, diagnosis, and management of infections in solid-organ transplant patients. AB - Infectious complications are still a significant cause of morbidity and death in solid-organ transplant patients, with significant infection being found in up to two-thirds of these individuals. The risk of infection in the organ transplant patient, particularly of opportunistic infection, is largely determined by 3 factors: the net state of immunosuppression, the epidemiologic exposures the patient encounters, and the consequences of the invasive procedures to which the patient is subjected. The most important principles of patient treatment are prevention, early diagnosis, and specific therapy. This issue is designed as a position paper by a group of experts on epidemiology, prevention, diagnosis, and management of infections in solid-organ transplant patients. We feel that our efforts may serve as an important first step in the development of guidelines in this area. PMID- 11389515 TI - Epidemiology of infections after solid-organ transplantation. AB - Infections are a major determinant of the outcome of solid-organ transplantation. Their incidence varies because of a number of factors. The type of organ transplanted, the degree of immunosuppression, the need for additional antirejection therapy, and the occurrence of technical or surgical complications all affect the incidence of infection after transplantation. There are 3 general time frames to consider. In the first month, bacterial and candidal infections predominate. From the second through the sixth months, infections "classically" associated with transplantation become manifest. Beyond the sixth month, the clinical course is largely determined by the presence or absence of chronic rejection. PMID- 11389516 TI - Pretransplant evaluation for infections in donors and recipients of solid organs. AB - The risk of infectious disease reactivation in recipients of and transmission by solid-organ transplants remains, and thorough screening and testing of recipient and donor is especially important. In conceiving screening strategies, it is crucial to consider the sensitivity and specificity of individual diagnostic tests in the context of their use. Furthermore, recognition of special risks for infectious complications of transplantation will help to guide preventive, diagnostic, and therapeutic steps in the control of infectious complications in individual patients. The acceptability of risks for infectious complications after transplantation depends also on the urgency of transplantation of a vital organ as well as the availability of organs. Although these principals are well accepted, standards for the extent of screening and criteria for inappropriate donors and exclusion of unfit recipients remain controversial to some extent. PMID- 11389518 TI - Posttransplant microbiological surveillance. AB - Posttransplant microbiological surveillance should be used when the likelihood of infection in a transplant recipient is high and the sensitivity and specificity of the test can provide a high positive or negative predictive value. Testing is also performed in some instances to monitor the patient's response to therapy. Examples of successful posttransplant microbiological surveillance include molecular detection of cytomegalovirus and Epstein-Barr virus and virus load determinations, as well as hepatitis B and C detection and virus load testing. Routine fungal and bacterial surveillance are generally not necessary, except for Candida colonization detection or vancomycin-resistant enterococcal detection in high-risk subgroups. The organ transplanted may also play a role in the type of routine surveillance recommended. PMID- 11389517 TI - Prophylactic measures in the solid-organ recipient before transplantation. AB - Pretransplant screening affords an important opportunity to detect and treat preexisting active infection in the solid-organ transplant recipient. In this article, pretransplant strategies for preventing infections after solid-organ transplantation are reviewed. In addition to the search for active preexisting infection in the transplant candidate, immunization remains a cornerstone of preventive practice. Because there is a suboptimal response to vaccinations in patients who are receiving immunosuppressive therapy, as well as in patients with end-stage organ dysfunction, standard immunization of the transplant candidate should be updated as early as possible in the course of the illness, including pneumococcal, influenza, and hepatitis B vaccines. Liver transplant candidates should receive hepatitis A vaccine, and children should receive Haemophilus influenzae type B conjugate vaccine. All nonimmune pretransplant patients should be considered candidates for the varicella vaccine. The management of special risk groups is discussed in detail. PMID- 11389519 TI - Prophylaxis strategies for solid-organ transplantation. AB - In addition to the net state of immunosuppression, the risk of infection after transplantation is largely determined by the transplant recipient's epidemiologic exposures. Potential sources of infection in the transplant recipient include the environment and the recipient's endogenous flora. This article presents aspects of prevention of infection after solid-organ transplantation such as avoidance of epidemiologic exposures, antibacterial prophylaxis, prophylaxis for tuberculin positive transplant recipients, and prophylaxis against infections with Pneumocystis carinii and Toxoplasma gondii. PMID- 11389520 TI - Management of cytomegalovirus infection and disease after solid-organ transplantation. AB - Cytomegalovirus (CMV) continues to be a cause of substantial morbidity and death after solid-organ transplantation. There are 3 major consequences of CMV infection: CMV disease, including a wide range of clinical illnesses; superinfection with opportunistic pathogens; and injury to the transplanted organ, possibly enhancing chronic rejection. This article discusses the considerable progress that has been made in elucidating risk factors for CMV disease, in the rapid detection of CMV in clinical specimens, and in the use of antiviral chemotherapy and immunoglobulin to prevent and treat CMV disease after solid-organ transplantation. PMID- 11389521 TI - Diagnosis and management of posttransplant lymphoproliferative disorder in solid organ transplant recipients. AB - The Epstein-Barr virus (EBV) has a pivotal pathophysiologic role in the development of most lymphoproliferative disorders that occur after solid-organ transplantation. The term "EBV-associated posttransplant lymphoproliferative disorder" (PTLD) includes all clinical syndromes of EBV-associated lymphoproliferation, ranging from uncomplicated posttransplant infectious mononucleosis to true malignancies that contain clonal chromosomal abnormalities. PTLDs are historically associated with a high mortality rate in patients who have a monoclonal form of the disorder. Recently described approaches to pathology, diagnosis, treatment, and preventive strategies of PTLD, however, have the potential to improve outcome. PMID- 11389522 TI - Prevention of fungal and hepatitis virus infections in liver transplantation. AB - Invasive fungal infections, especially those caused by Candida albicans, and recurrence of hepatitis B virus (HBV) and hepatitis C virus (HCV) infection after transplantation are common complications in orthotopic liver transplant (OLT) recipients. Candida species account for >50% of all invasive fungal infections, which occur in 10%--15% of OLT recipients. The epidemiology and pathogenesis of invasive fungal infections are unique to each type of organism. Fluconazole is effective and safe in the prevention of Candida infection after OLT. Preventive measures against Aspergillus or Cryptococcus remain ill defined. Both HBV and HCV recur almost universally after OLT in infected individuals. The natural course of HBV and HCV, leading to end-stage liver damage, is accelerated. In OLT patients, administration of immunoglobulin with high titers against HBV, alone and/or in combination with lamivudine, immediately after transplantation reduces the recurrence of HBV. The combination of interferon and ribavirin is mildly effective in OLT patients who have evidence of recurrent hepatitis, and additional alternatives are being evaluated. PMID- 11389523 TI - Management of urinary tract infections and lymphocele in renal transplant recipients. AB - The most frequent infectious complication after renal transplantation is urinary tract infection. This article deals with antimicrobial prophylaxis, treatment of early and relapsing urinary tract infections, and management of asymptomatic bacteriuria in renal transplant patients. The incidence of lymphocele after renal transplantation varies, and its treatment is still controversial. Management options are discussed. PMID- 11389524 TI - Epidemiology and management of infections after lung transplantation. AB - Lung transplantation has become an accepted treatment for end-stage pulmonary parenchymal and vascular diseases. Infections still are the most common cause of early and late morbidity and mortality in lung transplant recipients. Bacterial infections comprise approximately half of all infectious complications. Cytomegalovirus (CMV) infections and disease have become less frequent, because of prophylaxis with ganciclovir. Because CMV is also involved in the pathogenesis of obliterative bronchiolitis, the frequency of this infection may also reduce the occurrence of this main obstacle to successful lung transplantation. Invasive fungal infections remain a problem, but they have also decreased in frequency because of better control of risk factors such as CMV disease and preemptive antifungal therapy. Nonherpes respiratory viral infections have emerged as a serious problem. Their severity may be reduced by treatment with ribavirin. Meticulous postoperative surveillance, however, is still crucial for the management of lung transplant patients with respect to early detection and treatment of rejection and infection. PMID- 11389527 TI - Fatal connections: when DNA ends meet on the nuclear matrix. AB - A damaged nucleus has long been regarded simply as a "bag of broken chromosomes," with the DNA free ends moving around and forming connections with randomly encountered partners. Recent evidence shows this picture to be fundamentally wrong. Chromosomes occupy specific nuclear domains within which only limited movement is possible. In a human diploid nucleus, 6.6 x 10(9) base pairs (bp) of DNA are compartmentalized into chromosomes in a way that allows stringent control of replication, differential gene expression, recombination and repair. Most of the chromatin is further organized into looped domains by the dynamic binding of tethered bases to a network of intranuclear proteins, the so-called nuclear scaffold or matrix. Thus, DNA movement is severely curtailed, which limits the number of sites where interchanges can occur. This intricate organizational arrangement may render the genome vulnerable to processes that interfere with DNA repair. Both lower and higher eukaryotic cells perform homologous recombination (HR) and illegitimate recombination (IR) as part of their survival strategies. The repair processes comprising IR must be understood in the context of DNA structural organization, which is fundamentally different in prokaryotic and eukaryotic genomes. In this paper we first review important cellular processes including recombination, DNA repair, and apoptosis, and describe the central elements involved. Then we review the different DNA targets of recombination, and present recent evidence implicating the nuclear matrix in processes which can induce either repair, translocation, deletion, or apoptosis. J. Cell. Biochem. Suppl. 35:3-22, 2000. PMID- 11389529 TI - Signal transduction pathways and chromatin structure in cancer cells. AB - Molecular mechanisms controlling gene expression include cell shape, mechanical and chemical signal transduction pathways, chromatin remodeling, and DNA methylation. In this article, we will review the contribution of these molecular mechanisms and structural alterations in the malignant transformation of cells. The mechanical signaling pathway consists of the tissue matrix system that links together the three-dimensional skeletal networks, the extracellular matrix, cytoskeleton, and nuclear matrix. The cytoskeleton array is a dynamic system that transmits signals from the cell exterior to nuclear DNA. The composition and function of this mechanical signaling pathway is altered in cancer cells. Chemical signaling pathways such as the Ras/mitogen-activated protein kinase (MAPK) pathway stimulate the activity of kinases that modify transcription factors, histones, and chromatin remodeling factors. Oncoproteins deregulating this signaling pathway set in motion a series of events that cumulate to chromatin remodeling and aberrant gene expression. J. Cell. Biochem. Suppl. 35:27 35, 2000. PMID- 11389528 TI - DNA loop domain organization: the three-dimensional genomic code. AB - It is well known that aberrations in the nuclear matrix contribute to the development of cancer, but many aspects of this process remain unknown. The mammalian sperm nuclear matrix serves as a distinctive model of DNA loop domain organization by the nuclear matrix since the integrity of the DNA structure can be measured by the ability of the paternal chromosomes to participate in embryogenesis. The structure of the nuclear matrix is known to be important for normal cellular functions such as transcriptional regulation and DNA replication. Even small aberrations in DNA structural organization in the sperm cell could have disastrous consequences for the embryo if they were essential for function. Recent work from our laboratory suggests that sperm nuclei with disrupted nuclear matrix structures but intact DNA cannot participate fully in embryogenesis, suggesting that the structural organization of DNA may provide important, heritable information that is necessary for development. We term the DNA sequence together with its three-dimensional organization the "genomic code." We suggest that the sperm nucleus is an ideal model for understanding the principles of the involvement of the three-dimensional structure of DNA in normal cellular function. Finally, the implications for cancer about what we can learn using sperm DNA as a model about the "genomic code" are discussed. J. Cell. Biochem. Suppl. 35:23-26, 2000. PMID- 11389530 TI - Linking chromatin architecture to cellular phenotype: BUR-binding proteins in cancer. PMID- 11389531 TI - Linking Notch signaling, chromatin remodeling, and T-cell leukemogenesis. AB - Intercellular communication that controls the developmental fate of multipotent cells is commonly mediated by the Notch family of transmembrane receptors. Specific transmembrane ligands activate Notch receptors on neighboring cells inducing the proteolytic liberation and nuclear translocation of the intracellular domain of Notch (N(IC)). Nuclear N(IC) associates with a transcriptional repressor known as C-promoter binding factor/RBP-J kappa, suppressor of hairless, or LAG-1, converting it from a repressor into an activator. Through physical interactions with chromatin remodeling enzymes and potentially with components of the transcriptional machinery, N(IC) activates target genes that mediate cell fate decisions. As Notch1 is disrupted via a chromosomal translocation in a subset of human T-cell leukemia, leading to a truncated polypeptide resembling N(IC), deregulated chromatin remodeling and transcription may fuel uncontrolled cell proliferation in this hematopoietic malignancy. This review summarizes the mechanics of Notch signaling and focuses on prospective molecular mechanisms for how constitutively active Notch might derail nuclear processes as an initiating step in T-cell leukemogenesis. J. Cell. Biochem. Suppl. 35:46-53, 2000. PMID- 11389532 TI - Rearrangement of chromatin domains in cancer and development. AB - Both the accomplishment of developmental programs and neoplastic transformation are linked to changes in the long-range organization of chromatin, in particular, DNA loop domains. The development of new methods that allow the study of interactions between the bases of DNA loops and the proteins of the nuclear matrix will help our understanding of the molecular mechanisms in such changes. These methods should also allow the establishment of a fingerprint "signature" for many cancers that may serve for diagnostic purposes. J. Cell. Biochem. Suppl. 35:54-60, 2000. PMID- 11389533 TI - Chromatin and cancer: causes and consequences. AB - In this review, we discuss recent evidence implicating chromatin structure in the etiology of cancer. In particular, we present evidence indicating that inappropriate regulation of chromatin structure inhibits normal cell differentiation pathways and stimulates uncontrolled cell proliferation, with the outcome being oncogenesis. Such inappropriate chromatin structures arise as a consequence of (i) chromosomal rearrangements that fuse gene-specific activators with global co-regulators, drastically altering activator function; (ii) hypermethylation of tumor suppressor gene promoters, resulting in their inactivation; or (iii) mistargeted nuclear compartmentalization of growth-control genes and their regulators, resulting in the up- or down-regulation of such genes. How does chromatin silence genes? Recent results from model in vivo systems argues that chromatin can repress transcription at two levels: (i) by sterically interfering with the binding of transcription factors to the promoter, thereby blocking initiation; and (ii) at a step subsequent to the binding of activators and recruitment of the preinitiation complex. J. Cell. Biochem. Suppl. 35:61-68, 2000. PMID- 11389534 TI - Features of nuclear architecture that influence gene expression in higher eukaryotes: confronting the enigma of epigenetics. AB - Complex mechanisms that influence gene expression in mammalian cells have been studied intensively over recent years. Genetic elements that control both the tissue specific patterns and levels of gene expression together with the proteins they bind have been characterised in detail and are clearly pivotal in activating pathways of gene expression. But it is also clear that the behaviour of these genetic elements is complicated by epigenetic factors, so that their introduction into cells with the necessary developmental history-and hence appropriate global concentrations of essential transcription factors-will not guarantee the desired levels of transcription. Recent experiments have reinforced this view and confirmed that apparently critical functions performed by defined genetic elements at certain chromosomal sites are not inevitably recapitulated at other chromosomal locations. Hence, a re-evaluation of the function of critical control elements is required using experimental systems that simplify the range of factors arising from local chromatin organisation. In this way, it should be possible to reveal the intricacies of gene expression that might eventually allow us to reproduce natural levels of expression from artificial gene constructs in human cells. J. Cell. Biochem. Suppl. 35:69-77, 2000. PMID- 11389535 TI - DNA methylation, nuclear structure, gene expression and cancer. AB - DNA methylation, chromatin structure, transcription, and cancer have traditionally been studied as separate phenomena. Recent data provide now direct physical and functional links between these processes revealing a complex network of interactions and mutual dependences. Methylated DNA is bound by methyl-CpG binding protein (MeCP) complexes that include histone deacetylases (HDACs). This recruitment of HDACs is suggested to promote local chromatin condensation and thereby repress gene expression. Most recently, also complexes of DNA methyltransferase (Dnmt1) with transcriptional repressors, DMAP1 and pRB, have been described providing a direct link to transcriptional regulation and tumor suppression. Inactivation of the DNA methyltransferase genes (Dnmt1, 3a, and 3b) was found to be lethal in mice and several human diseases (ICF and Rett syndrome) turned out to be linked to DNA methylation. In particular, global hypomethylation has been found in tumor samples together with cancer-type-specific, local hypermethylation. Taken together, these lines of evidence clearly underscore the central role of DNA methylation in the regulation of gene expression and chromatin structure during normal development and diseases like cancer. J. Cell. Biochem. Suppl. 35:78-83, 2000. PMID- 11389536 TI - Subnuclear organization and trafficking of regulatory proteins: implications for biological control and cancer. AB - The regulated and regulatory components that interrelate nuclear structure and function must be experimentally established. A formidable challenge is to define further the control of transcription factor targeting to acceptor sites associated with the nuclear matrix. It will be important to determine whether acceptor proteins are associated with a pre-existing core-filament structural lattice or whether a compositely organized scaffold of regulatory factors is dynamically assembled. An inclusive model for all steps in the targeting of proteins to subnuclear sites cannot yet be proposed. However, this model must account for the apparent diversity of intranuclear targeting signals. It is also important to assess the extent to which regulatory discrimination is mediated by subnuclear domain-specific trafficking signals. Furthermore, the checkpoints that monitor subnuclear distribution of regulatory factors and the sorting steps that ensure both structural and functional fidelity of nuclear domains in which replication and expression of genes occur must be biochemically and mechanistically defined. There is emerging recognition that placement of regulatory components of gene expression must be temporally and spatially coordinated to facilitate biological control. The consequences of breaches in nuclear structure-function relationships are observed in an expanding series of diseases that include cancer [Weis et al., 1994; Rogaia et al., 1997; Yano et al., 1997; Rowley, 1998; Zeng et al., 1998; McNeil et al., 1999; Tao and Levine, 1999a] and neurological disorders [Skinner et al., 1997]. As the repertoire of architecture-associated regulatory factors and cofactors expands, workers in the field are becoming increasingly confident that nuclear organization contributes significantly to control of transcription. To gain increased appreciation for the complexities of subnuclear organization and gene regulation, we must continue to characterize mechanisms that direct regulatory proteins to specific transcription sites within the nucleus so that these proteins are in the right place at the right time. J. Cell. Biochem. Suppl. 35:84-92, 2000. PMID- 11389537 TI - Alterations in subnuclear trafficking of nuclear regulatory factors in acute leukemia. AB - The nuclear matrix plays an important role in the functional organization of the nucleus in part by locally concentrating regulatory factors involved in nucleic acid metabolism. A number of nuclear regulatory proteins initially identified due to their involvement in human cancer are localized to discrete nuclear matrix attached foci and correct nuclear partitioning likely plays a role in their function. Two such examples are promyelocytic leukemia (PML) and acute myelogenous leukemia-1 (AML-1; Runx1). PML, the target of the t(15;17) in acute PML, is localized to PML nuclear bodies (also termed Nuclear Domain 10 and PML oncogenic domains), a nuclear matrix-associated body whose function appears to be quite complex, with probable roles in cancer, apoptosis, and in acute viral infections. In t(15;17)-containing leukemic cells, the PML nuclear bodies are disrupted, but reform when the leukemic cells are induced to differentiate in the presence of all-trans retinoic acid. AML1 (RUNX1) is a key regulator of hematopoietic differentiation and AML1 proteins are found in nuclear compartments that reflect their roles in transcriptional activation and repression. The t(8;21), associated with AML, results in a chimeric transcription factor, AML 1/ETO (eight twenty one), that remains attached to the nuclear matrix through targeting signals contained in the ETO protein. When co-expressed, ETO and AML 1/ETO co-localize to a nuclear compartment distinct from that of AML1 or PML nuclear bodies. Interestingly, enforced expression of ETO or AML-1/ETO changes the average number of PML nuclear bodies per cell. Thus, chromosomal translocations involving AML1 result in altered nuclear trafficking of the transcription factor as well as other changes to the nuclear architecture. J. Cell. Biochem. Suppl. 35:93-98, 2000. PMID- 11389538 TI - Subnuclear dynamics and transcription factor function. AB - At a simplistic level, the nucleus can be thought of as singular organelle with a nuclear envelope designed to isolate the biochemical reactions required for gene transcription and DNA replication from the cytoplasm. It has become increasingly clear, however, that many higher levels of organization exist within the nucleus. A functional consequence of this organization is that nuclear processes that include transcription, RNA processing, and DNA synthesis are isolated to specific intranuclear domains to ensure efficiency. With the advent of GFP technologies and increasingly sophisticated instrumentation, we have continued to dissect the relationship between organization and function, in particular using live cells and ligand-dependent steroid receptors as a model system. These new opportunities have provided further insight into receptor function and the dependence upon intranuclear dynamics that take place within minutes of hormone addition. J. Cell. Biochem. Suppl. 35:99-106, 2000. PMID- 11389539 TI - The dynamics of acentric chromosomes in cancer cells revealed by GFP-based chromosome labeling strategies. AB - Autonomous replicons, such as viral episomes and oncogene containing double minute chromosomes (DMs), lack centromeres and consequently should be lost rapidly when the nuclear membrane breaks down at mitosis. Surprisingly, they are not. This raises the important question of the mechanisms that enable their efficient transmission to daughter cells. We review recent developments in GFP based chromosome labeling strategies that enable real time analyses using high resolution light microscopy to provide insights into this issue. The results reveal that episomes and DMs both adhere to host chromosomes, a process referred to as "chromosome tethering". Such association enables acentric molecules to use the chromosomal centromere in trans, thereby achieving efficient transmission to daughter cells. This unique mechanism of mitotic segregation also raises the possibility of developing a new class of anti-cancer drugs that work by selectively eliminating growth enhancing genes from cancer cells. J. Cell. Biochem. Suppl. 35:107-114, 2000. PMID- 11389540 TI - Mutant p53: "gain of function" through perturbation of nuclear structure and function? AB - Mutant p53 not simply is an inactivated tumor suppressor, as at least some mutant p53 proteins exhibit oncogenic properties. Mutant p53 thus is the most commonly expressed oncogene in human cancer. Accordingly, the expression of mutant p53 in tumors often correlates with bad prognosis, and expression of mutant p53 in p53 negative tumor cells enhances their transformed phenotype. The molecular basis for this "gain of function" is not yet understood. However, the finding that mutant p53 tightly associates with the nuclear matrix in vivo, and with high affinity binds to nuclear matrix attachment region (MAR) DNA in vitro, suggests that these activities are connected and may result in perturbation of nuclear structure and function in tumor cells. MAR-binding of mutant p53 most likely is due to conformation-selective DNA binding by mutant p53, i.e. the specific interaction of a given mutant p53 protein with regulatory or structural genomic DNA elements that are able to adopt specific non-B-DNA conformations. In support to this assumption, human mutant p53 (Gly(245)-->Ser) was shown to bind to repetitive DNA elements in vivo that might be part of MAR elements. This further supports a model according to which mutant p53, by interacting with key structural components of the nucleus, exerts its oncogenic activities through perturbation of nuclear structure and function. J. Cell. Biochem. Suppl. 35:115 122, 2000. PMID- 11389541 TI - Evidence for lectin signaling to the nuclear matrix: cellular interpretation of the glycocode. PMID- 11389542 TI - Significance of protein kinase CK2 nuclear signaling in neoplasia. AB - Many stimuli play a role in influencing the structure and function of chromatin and nuclear matrix through post-translational modifications of the component proteins in these dynamic structures. We propose that the protein serine/threonine kinase CK2 (formerly casein kinase II) is one such agent that is involved in signal transduction in the nuclear matrix and chromatin in response to a variety of stimuli. Protein kinase CK2 appears to undergo rapid modulations in its association with nuclear matrix and nucleosomes in response to mitogenic signals and is involved in the phosphorylation of a variety of intrinsic proteins in these structures depending on the state of genomic activity. In addition, its association or loss from the nuclear matrix may also influence the apoptotic activity in the cell. CK2 has been found to be dysregulated in virtually all the neoplasias examined and nuclear association appears to be an important facet of its expression in tumor cells. We hypothesize that CK2 provides a functional paradigm linking the nuclear matrix and chromatin structures. Identification of precise loci of action of CK2 in these structures and how they influence the morphological appearance of the nucleus under normal and abnormal growth conditions would be an important future direction of investigation. J. Cell. Biochem. Suppl. 35:130-135, 2000. Published 2001 Wiley-Liss, Inc. PMID- 11389543 TI - Nuclear matrix proteins as cancer markers. AB - The transformation of normal cells to a malignant state has long been detected by light microscopy as visible changes in nuclear morphology. These changes include abnormal nuclear shape, increased nuclear to cytoplasmic ratio, and the presence of additional and abnormal nucleoli. Metaplasia,dysplasia, carcinoma-in-situ, and gross malignant tumors are diagnosed and graded pathologically by this traditional method. The resulting relative increase in DNA concentration within these cells produces a greater affinity for Hematoxilyn and Eosin staining, and thus, the characteristic blue color of cancerous tissues. As understanding of the cell structure expanded, the nuclear matrix emerged as an integral component of genetic processing and therefore, became an important cellular entity for study of malignant transformation. Also, several types of cancer have revealed discreet alterations in their respective nuclear matrices. One potential application of these nuclear matrix changes is development of detection and monitoring tests that would reveal the presence of abnormal cells. These tests could be utilized at a number of points in the disease process including prior to gross physical symptoms, and thereby significantly reduce patient morbidity and mortality. A second potential application of the nuclear matrix is to utilize it as a tissue specific protein targeting system to address narrowly directed therapeutic treatments, and thereby avoid the systemic side effects from broad-spectrum therapies like radiation. This paper addresses the role of the nuclear matrix in both normal cells and transformed cells, and highlights several research efforts that have advanced the ability to detect, track, and potentially treat neoplasms at the molecular level. J. Cell. Biochem. Suppl. 35:136-141, 2000. PMID- 11389544 TI - Changes in sub-nuclear structures and functional perturbations: implications for radiotherapy. AB - The eukaryotic cell nucleus is required to accomplish its functions (e.g., replicating transcription, DNA repair, hmRNA processing, etc.) within the context of a highly organized structure [Wei X, Samarabandu J, Devdhar RS, Siegel AJ, Acharya R, Berezney R. 1998. Science 281:1502-1506.], since many cancer therapeutic modalities utilize the nucleus as target for a cytotoxic outcome. A better understanding of the organizational disruption of sub-nuclear structures and subsequent loss of nuclear function is the key to knowing both the mechanism of action of, and the basis of cellular sensitivity to, therapeutic agents such as ionizing radiation. With this prospect, we examine four examples in which changes in specific nuclear structures or functions lead to significant therapeutic end points, e.g. cell death, radiosensitization, or the intrinsic radioresistance of tumor cells. The inter-relationships delineated in these examples provide a paradigm that delineates a relationship between disruption of nuclear organization, loss of function and a point of intervention that affects a therapeutic outcome. The examples specifically address issues related to radiation and thermal therapy. However, the concepts that result from these studies are translatable to other cancer therapeutic modalities. In addition, the results echo a basic principle that proper nuclear organization is critical to the maintenance of cellular viability and genomic stability. J. Cell. Biochem. Suppl. 35:142-150, 2000. PMID- 11389545 TI - Quantitative nuclear grade (QNG): a new image analysis-based biomarker of clinically relevant nuclear structure alterations. AB - This review addresses the potential clinical value of using quantitative nuclear morphometry information derived from computer-assisted image analysis for cancer detection and predicting outcomes such as tumor stage, recurrence, and progression. Today's imaging technology uses sophisticated hardware platforms coupled with powerful and user-friendly software packages that are commercially available as complete image analysis systems. There are many different mathematically derived nuclear morphometric descriptors (NMD's) (i.e. texture features) that can be calculated by these image analysis systems, but for the most part, these NMD's quantify nuclear size, shape, DNA content (ploidy), and chromatin organization (i.e. texture, both Markovian and non-Markovian) parameters. We have utilized commercially available image analysis systems and the NMD's calculated by these systems to create a mathematical solution, termed quantitative nuclear grade (QNG), for making clinical, diagnostic, and prognostic outcome predictions in both prostate and bladder cancer. A separate computational model is calculated for each outcome of interest using well-characterized and robust training, testing, and validation patient sample sets that adequately represent the selected population and clinical dilemma. A specific QNG solution may be calculated either by non-parametric statistical methods or non-linear mathematics employed by artificial neural networks (ANNs). The QNG solution, a measure of genomic instability, provides a unique independent variable to be used alone or to be included in an algorithm to assess a specific clinical outcome. This approach of customization of the nuclear morphometric descriptor (NMD) information through the calculation of a QNG solution mathematically adjusts for redundancy of features and reduces the complexity of the inputs used to create decision support tools for patient disease management. J. Cell. Biochem. Suppl. 35:151-157, 2000. PMID- 11389546 TI - Decidualization and implantation: embryo-uterine bioinformatics at work. AB - The implantation of the blastocyst into a nurturing endometrium involves two overlapping steps: 1. The blastocyst-endometrial luminal epithelial attachment. 2. The decidualization of the endometrial stroma. An intriguing question is how does the blastocyst identify the uterine implantation site. Current research is focused on hypothetical soluble signaling molecules released by the blastocyst for conditioning a discrete uterine luminal epithelial domain for implantation. A still unresolved issue is the functional significance of receptor autophosphorylation following binding of uterine epithelial cell-derived heparin binding epidermal growth factor-like growth factor to the epidermal growth factor receptor on trophoectodermic cell surfaces. With recent results hinting at the role of signaling proteins associated with the bone morphogenetic protein, fibroblast growth factor, WNT and hedgehog families to enable embryo implantation, the dynamics of uterine-embryo interaction becomes linked to fundamental cellular pathways of growth, differentiation and apoptosis. PMID- 11389547 TI - Ovarian expression of melatonin Mel(1a) receptor mRNA during mouse development. AB - Melatonin, secreted by the pineal gland, is involved in the regulation of many physiological functions of various species of animals. In the present study, the expression of gene for melatonin Mel(1a) receptor (MelR) was evaluated in the ovary, hypothalamus, and pituitary according to the developmental stages in female mice. Semiquantitative reverse-transcription polymerase chain reaction (RT PCR) and in situ PCR techniques were applied. According to the developmental stages, gene for MelR was differently expressed on ovary, hypothalamus, and pituitary. MelR gene was first expressed on pituitary prior to the expression in hypothalamus and ovary. Ovarian MelR gene started to express at birth. Unlike hypothalamic expression of MelR gene which was identified after birth, in pituitary, it was expressed at 16 days post coitum. In the ovary, the expression signal of MelR gene was identified on granulosa cells. However, the signal was not detected in the theca cells. It was weak in the primordial and atretic follicles. Taken together, it can be considered that melatonin has a pivotal role in the folliculogenesis. PMID- 11389548 TI - Primary structure and developmental expression of Dp ZP2, a vitelline envelope glycoprotein homolog of mouse ZP2, in Discoglossus pictus, one of the oldest living Anuran species. AB - A glycoprotein of the Xenopus vitelline envelope, gp 69/64, which mediates sperm binding, is closely related to the components of ZPA family, such as the mouse zona pellucida ZP2. To test the generality of these findings, we studied Discoglossus pictus, a species evolutionary distant from Xenopus and identified as a protein of 63 kDa in the vitelline envelope. Preliminary studies suggest that this protein may bind sperm at fertilization. We found that the 63-kDa protein is glycosylated and contains both N- and O-linked chains. We have cloned the cDNA encoding the Discoglossus protein of 63 kDa (Dp ZP2) by screening a Discoglossus cDNA library using Xenopus gp 69/64 cDNA as a probe. Analysis of the deduced sequence of Discoglossus protein revealed 48% identity with Xenopus gp 69/64 and 37-40% identity with mouse ZP2. The sequence conservation included a ZP domain, a potential furin cleavage site and a putative transmembrane domain. The N-terminus region of Dp ZP2 was 40% identical to the corresponding region of Xenopus gp 69/64 which has been shown to be essential for sperm binding to the VE. Although, as of yet, there is no evidence for sperm binding at the Dp ZP2 N terminus, it is interesting that in this region three potential O-glycosylation sites are conserved in both species, in contrast to N-glycosylation sites. It was found that the Dp ZP2 mRNA is expressed in stage 1 oocytes and in the follicle cells surrounding the oocyte. Similarly, in Xenopus oocytes, the gp 69/64m RNA, was found in the oocytes, as well as in the somatic cells. Mol. Reprod. Dev. 59:133-143, 2001. PMID- 11389549 TI - Expression of genes involved in mammalian meiosis during the transition from egg to embryo. AB - The ooplasm of higher eukaryotes provides substances necessary for completing the last stages of meiosis and initiating the first mitotic division. These processes are firmly attuned to other events in the egg and newly formed embryo, such as switching from the use of maternal transcripts to the onset of zygotic transcription. In mammals little is known about the molecular mechanisms guiding this transition, largely due to the lack of information about genes expressed in the egg and early embryos. Studies of yeast mitosis have contributed much of what is known about the vertebrate cell cycle, and recent reports indicate that homologs of yeast DNA repair genes also function during mammalian gametogenesis. To examine whether this conservation can be expanded to include genes operative in oocyte meiosis, we performed a computer-based search for homologs of yeast genes that are induced during sporulation in C. elegans, Drosophila, and mammals. Results from this study suggest that yeast and higher eukaryotes share genes that coordinate the overall process of meiosis. However intriguing differences exist, reflecting the distinctive mechanisms governing the progression of meiosis in each organism. ESTs representing more than half of the mammalian homologs are present in mouse cDNA libraries that contains genes controlling the meiosis/mitosis transition. About 50% of these genes contain potential cis elements for cytoplasmic polyadenylation in their 3'-UTR, suggesting the importance of controlled translation in the egg and zygote. PMID- 11389550 TI - Tissue inhibitor of matrix metalloproteinase-3 expression in the mouse uterus during implantation and artificially induced decidualization. AB - During implantation in mice, tissue inhibitor of matrix metalloproteinases-3 is believed to play a key role in inhibiting matrix metalloproteinase activity associated with embryo invasion and tissue remodeling. The first objective of this study was to quantitatively compare the steady-state mRNA levels of tissue inhibitors of matrix metalloproteinases between segments of the mouse uterus undergoing decidualization compared to those that are not during early pregnancy plus oil-induced decidualization. Steady-state tissue inhibitor of metalloproteinase-3 mRNA levels were significantly greater in implantation compared to interimplantation areas on days 6 and 7 of pregnancy and in stimulated compared to nonstimulated uterine horns at 48 and 72 hr after artificial induction of decidualization. Steady-state tissue inhibitor of metalloproteinase-1 mRNA levels were significantly greater in implantation compared to interimplantation areas on days 5-8 of pregnancy and in stimulated compared to nonstimulated uterine horns at 24, 48, and 72 hr after oil stimulation. Therefore, the steady-state mRNA levels of tissue inhibitors of metalloproteinase-1 and -3 increased in the uterus during decidualization. The second objective of this study was to determine if transforming growth factor beta1 influences tissue inhibitors of metalloproteinase mRNA concentrations in mouse endometrial stromal cells. As determined by Northern blot analyses, transforming growth factor beta1 significantly increased tissue inhibitors of matrix metalloproteinases-1 and -3 mRNA levels in cultured mouse endometrial stromal cells isolated from uteri sensitized for decidualization. On the other hand, interleukin-1, epidermal growth factor, and leukemia inhibitory factor had no effect. The results of this study further characterize the tissue inhibitor of metalloproteinase expression in the uterus during implantation and artificially induced decidualization and the potential control of their expression in the stroma by transforming growth factor. PMID- 11389551 TI - alpha-, beta-, gamma-catenin, and p120(CTN) expression during the terminal differentiation and fusion of human mononucleate cytotrophoblasts in vitro and in vivo. AB - The cadherins play key roles in the formation and organization of the mammalian placenta by mediating cellular interactions and the terminal differentiation of trophoblastic cells. Although cadherin function is regulated by the cytoplasmic proteins, known as the catenins, the identity and expression pattern(s) of the catenins present in the trophoblastic cells of the human placenta have not been characterized. In these studies, we have determined that alpha-, beta-, gamma catenin, and p120(ctn) expression levels are high in villous cytotrophoblasts isolated from the human term placenta but decline as these cells undergo aggregation and fusion to form syncytium with time in culture. In contrast, the expression levels of these four catenin subtypes remained constant in non-fusing JEG-3 choriocarcinoma cells at all of the time points examined in these studies. alpha-, beta-, gamma-catenin, and p120(ctn) expression was further immunolocalized to the mononucleate cells present in these two trophoblastic cell cultures. Similarly, intense immunostaining for all four catenins was detected in the mononucleate villous cytotrophoblasts of the human first trimester placenta. Collectively, these observations demonstrate that the expression levels of alpha , beta-, gamma-catenin, and p120(ctn) are tightly regulated during the formation of multinucleated syncytium in vitro and in vivo. PMID- 11389552 TI - Ovarian expression of human insulin-like growth factor-I in transgenic mice results in cyst formation. AB - Insulin-like growth factor-I (IGF-I) has been implicated in a wide variety of physiological processes including ovarian function. To better understand the ovarian role of IGF-I, transgenic mice harbouring a human IGF-I cDNA (hIGF-I) under the control of the mouse LH receptor promoter were generated. Expression of the hIGF-I, determined by Northern blot, was found to occur in the gonad tissues of these transgenic mice. The hIGF-I protein was also detectable by radioimmunoassay in ovarian extracts as well as in the plasma. The fertility of mating transgenic females, as estimated by the number of implantation sites post coitum, did not appear to be affected. However, transgenic females who failed to mate and produce offspring were found to possess polycystic ovaries. Evaluation of testosterone, estradiol, and LH levels revealed that transgenic animals had significantly elevated circulating levels of testosterone compared to their non transgenic littermates, while LH levels in transgenic females were significantly lower. Yet, estradiol appeared to be unaffected. These results support the contention that the IGF system plays an important role in ovarian function and that an imbalance in this system may result in ovarian pathology. PMID- 11389553 TI - Haptoglobin inhibits lecithin-cholesterol acyltransferase in human ovarian follicular fluid. AB - The activity of the enzyme lecithin-cholesterol acyltransferase (LCAT; E.C. 2.3.1.43) is involved in the removal of cholesterol excess from peripheral cells. This activity is stimulated by the HDL (high density lipoprotein) apolipoprotein A1 (ApoA1). Haptoglobin (Hpt) was previously found to be associated with ApoA1 in ovarian follicular fluid. LCAT activity was analyzed in follicular fluids, collected from an IVF program, containing different amounts of Hpt or Hpt/ApoA1 ratio. Addition of purified Hpt to follicular fluid caused a decrease in the enzyme activity, which was measured as the rate of synthesis of cholesteryl esters. In the fractions of fluid proteins, as obtained by gel filtration chromatography, Hpt and HDL were titrated by ELISA while the LCAT activity was assayed by using radioactive cholesterol and purified HDL. When isolated LCAT was incubated with fractions containing different Hpt/ApoA1 ratios, the enzyme activity was found negatively correlated with the Hpt/ApoA1 ratio (P < 0.01). LCAT kinetic parameters were measured in two fractions with the same amount of ApoA1 (5 microg/ml) but different amounts of Hpt (0.69 or 3.77 microg/ml): the V(max) did not change while the K(m) values were 24.1 or 78.6 microM in the presence of the low or high Hpt level, respectively. The analysis of fluids associated with cytoplasmically mature MII oocytes, in a cross-sectional study, confirmed that a negative correlation exists between the Hpt/ApoA1 ratio and the LCAT activity (P < 0.01). The results suggest that Hpt inhibits the reverse transport of cholesterol by preventing ApoA1 stimulation of the LCAT activity. PMID- 11389555 TI - Requirement of cyclin B2, but not cyclin B1, for bipolar spindle formation in frog (Rana japonica) oocytes. AB - Cyclin B, the regulatory subunit of maturation-promoting factor (MPF), comprises several subtypes that are presumed to confer different functions on MPF although no direct evidence has been provided to date. To clarify the difference in the roles of cyclins B1 and B2, we used frog (Rana japonica) oocytes in which MPF is formed only after progesterone stimulation because it is possible to produce oocytes containing either cyclin B1-MPF or cyclin B2-MPF by antisense RNA mediated translational inhibition of each mRNA. Using this advantage, we investigated the functions of cyclins B1 and B2 and obtained the following results: (a) oocytes synthesizing cyclin B2-MPF underwent meiosis I and II with formation of a bipolar spindle at each metaphase; (b) oocytes synthesizing cyclin B1-MPF formed a monopolar spindle at metaphase I and extruded an abnormal polar body; and (c) both oocytes underwent germinal vesicle breakdown (GVBD) and chromosome condensation. Immunocytochemical observations also revealed continuous localization of cyclin B2 on the spindle during meiosis. These results provide evidence of the requirement of cyclin B2, but not cyclin B1, for organizing the bipolar spindle, though either cyclin B1 or B2 is redundant for inducing GVBD and chromosome condensation. PMID- 11389554 TI - Cytoplasmic changes in relation to nuclear maturation and early embryo developmental potential of porcine oocytes: effects of gonadotropins, cumulus cells, follicular size, and protein synthesis inhibition. AB - Morphological and biochemical changes indicative of cytoplasmic maturation in relation to nuclear maturation progression and early embryo developmental potential was studied. Fluorescently labeled microfilaments and cortical granules were visualized by using laser scanning confocal microscopy. The mitogen activated protein (MAP) kinase phosphorylation and cyclin B1 levels were revealed by Western blot. With the maturation of oocytes, cortical granules and microfilaments were localized at the cell cortex. A cortical granule-free domain (CGFD) and an actin-thickening area were observed over both the MII spindle of a mature oocyte and chromosomes of a nocodazole-treated oocyte, suggesting that chromosomes, but not the spindle, determined the localization of CGFD and actin thickening area. In oocytes that are incompetent to resume meiosis, as indicated by the failure of germinal vesicle breakdown (GVBD), peripheral localization of cortical granules and microfilaments, phosphorylation of MAP kinase and synthesis of cyclin B1 did not occur after 44 hr in vitro. These cytoplasmic changes were also blocked when GVBD of meiotically competent oocytes was inhibited by cycloheximide. Culture of oocytes in a chemically defined medium showed that biological factors such as gonadotropins, cumulus cells and follicle size affected both nuclear and cytoplasmic maturation as well as embryo developmental potential. Absence of gonadotropins or removal of cumulus cells alone did not significantly influence GVBD or cyclin B1 levels, but decreased the final maturation and developmental ability of oocytes. A combination of gonadotropin absence and cumulus removal decreased GVBD, MAP kinase phosphorylation and embryo development. A high proportion of oocytes derived from small follicles were able to resume meiosis, synthesize cyclin B(1), phosphorylate MAP kinase and translocate CGs, but their maturation and embryo developmental ability were limited. Removal of cumulus cells from small follicle-derived oocytes severely affected their ability to undergo cytoplasmic and nuclear maturation. PMID- 11389556 TI - Caspase activity in newt spermatogonial apoptosis induced by prolactin and cycloheximide. AB - We previously showed in vivo and in vitro, that among the spermatogenic stages of the newt, prolactin (PRL) induces apoptosis specifically in the penultimate stage of secondary spermatogonia. In the current report, we demonstrate in vitro that cycloheximide (CHX), an inhibitor of protein synthesis, induces morphological apoptotic changes similar to those caused by PRL, such as chromatin condensation and apoptotic body formation. Next, we found that Z-VAD-fmk, an inhibitor of various caspases, suppressed the apoptosis induced by PRL and CHX, but ICE inhibitor Ac-YVAD-CHO or caspase-3 inhibitor Ac-DEVD-CHO did not. As high caspase activity was present in extracts of testes treated with CHX, we suggest that an unidentified caspase induces the morphological changes of apoptosis in newt spermatogonia. PMID- 11389557 TI - Germinal vesicle materials are not required for the activation of MAP kinase in porcine oocyte maturation. AB - The requirement of the germinal vesicle (GV) for the normal kinetics of mitogen activated protein (MAP) kinase activity during porcine oocyte maturation was investigated. Porcine follicular oocytes were enucleated, and the locations of their extracellular signal-regulated kinases 1 and 2 (ERK1/2), major MAP kinases in maturating porcine oocytes, were detected by indirect immunofluorescent microscopy. The MAP kinase activity was assayed as myelin basic protein (MBP) kinase activity, and the phosphorylation states of ERK1/2 were detected by immunoblotting analyses. Translocation of MAP kinase into the GV and association with the spindle were observed in intact oocytes, while MAP kinase in enucleated oocytes was distributed almost uniformly in cytoplasm throughout the culturing period. The phosphorylation and the activation of MAP kinase were induced, and the activity was comparable with that of control denuded oocytes. The high level of activity was maintained through maturation, even in the absence of spindle formation. These results indicate that the presence of nuclear material and translocation into the GV are dispensable for the activation of MAP kinase and that associating with the spindle is not required for maintenance of its activity though porcine oocyte maturation. PMID- 11389558 TI - Essential role of the nonreducing terminal alpha-mannosyl residues of the N linked carbohydrate chain of bovine zona pellucida glycoproteins in sperm-egg binding. AB - It has been proposed that mammalian sperm bind species-specifically to carbohydrate chains of zona pellucida glycoproteins at fertilization. Although the sperm ligand carbohydrate chains have been characterized in mice and pigs, the existence of the ligands of other mammals remains unclear. In order to explore the bovine sperm ligand, two in vitro competition assay methods were applied. As a result, a high-mannose-type carbohydrate chain, Manalpha1 6(Manalpha1-3)Manalpha1-6(Manalpha1-3)Manbeta1-4GlcNAcbeta1-4GlcNAc, which is the major neutral chain in bovine egg zona glycoproteins, was shown to possess bovine sperm ligand activity. When nonreducing terminal alpha-mannosyl residues were eliminated from the zona glycoproteins by alpha-mannosidase digestion, the ligand activity was reduced, indicating that the alpha-mannosyl residues play an essential role in bovine sperm-egg binding. The number of sperm binding to eggs was reduced to about one-half after fertilization. The ligand-active high-mannose type chain may be buried after fertilization, since its amount remains unchanged. Pretreatment of bovine sperm with the sperm ligand-carbohydrate chain significantly inhibited penetration of the sperm into oocyte and the male pronucleus formation. Thus, a correlation between the sperm ligand activity and in vitro fertilization rate was observed. PMID- 11389559 TI - Porcine oocyte activation: differing roles of calcium and pH. AB - Intracellular pH has recently been shown to increase during parthenogenetic activation of the porcine oocyte. In the following set of experiments, intracellular pH was monitored during activation and pronuclear development was assessed following activation treatments with calcium, in the absence of calcium, and in oocytes loaded with the calcium chelator BAPTA-AM in calcium-free medium. Intracellular pH increase was not different among groups when treating with 7% ethanol or 50 microM calcium ionophore, or during treatment with thimerosal for 12 or 25 min. Activation with thimerosal (200 microM, 12 min) followed by 8 mM dithiothreitol (DTT, 30 min) resulted in a decreased pronuclear development in calcium-free medium with or without BAPTA-AM loaded oocytes as compared to controls. Activation with 50 microM calcium ionophore resulted in pronuclear development that was different between the calcium-free and BAPTA-AM loaded oocytes in calcium-free medium. Similar incidences of pronuclear formation were observed in all ethanol treatment groups. It was concluded that external calcium as well as large changes in intracellular free calcium are not necessary for the increase in intracellular pH, but normal intracellular calcium signaling is critical for normal levels of pronuclear development. Finally, oocytes were measured for intracellular pH changes for 30 min following subzonal sperm injection. Intracellular pH did not increase, although pronuclear formation was observed 6 hr post SUZI. This suggested that major differences were still present between sperm-induced and parthenogenetic activation of the porcine oocyte. PMID- 11389561 TI - Living with support in a home in the community: predictors of behavioral development and household and community activity. AB - The purpose of this article was to review studies of behavioral development and household and community activity among adults with mental retardation living in community residential services and to distill knowledge about the factors that influence outcome. Research points to behavioral development occurring across the full spectrum of disability but influenced by mental retardation syndrome and the acquisition of pivotal skills. However, engagement in household and community activities has been found to be strongly related to individual adaptive behavior. People with more severe mental retardation are vulnerable to leading lives characterized by underoccupation and lack of community involvement. Moving from institutional to community-based residential services may be accompanied by significant increases in adaptive behavior, but a plateau effect on subsequent development has also been reported. There is substantial evidence to suggest that key aspects of effective teaching technology may be absent in community-based residential environments. Community settings support greater engagement in household and community activities than institutions. Ordinary housing stock and normative architecture and standards of material enrichment are to be preferred. The use of normative housing constrains group living to relatively small scale, but there is little evidence that smaller size within this range is to be preferred to larger size. There is little evidence to suggest that higher staff to-resident ratios lead to uniformly better outcomes, but staff orientation, working methods, and performance are important influences. Little is known about what precise characteristics of community location give rise to greater community integration. MRDD Research Reviews 7:75-83, 2001. PMID- 11389562 TI - Transition to adulthood: supporting young adults to access social, employment, and civic pursuits. AB - "Transition support" is a concept new to the field of secondary transition. This paper describes the development of a model of transition support as derived from the empirical literature (Hughes and Carter. [2000] The Transition Handbook. Baltimore, MD: Paul H. Brookes). The components of the Transition Support Model are both empirically based and field-tested among practitioners. The over 500 transition support strategies that compose the model have been used by practitioners in the field to improve educational and postschool outcomes for high school students. These strategies are grouped according to two main goals (i.e., "developing support in the environment" and "increasing students' competence") and corresponding components. Literature supporting the Transition Support Model is described as well as the use of the model in the transition planning process. Suggestions are made for application of transition support in high schools and the need for future research. MRDD Research Reviews 7:84-90, 2001. (c) 2001 Wiley-Liss, Inc. PMID- 11389563 TI - Living with support in the community: predictors of choice and self determination. AB - Ensuring that people with intellectual disability experience typical levels of choice and self-determination has become an essential objective for quality disability services. Three perspectives on self-determination were identified in the literature: psycho-educational, ecological, and socio-political. Personal characteristics, specific self-determination competencies, and environmental variables all were found to be associated with choice and self-determination. Living environments that were smaller and more individualized were linked to greater choice and self-determination. The theoretical perspective investigators applied to the issues influenced approaches to research and intervention, and differences in emphasis were evident regarding the relative importance of self determination competencies and environmental factors. MRDD Research Reviews 7:91 98, 2001. PMID- 11389564 TI - Living with support in the community: predictors of satisfaction with life. AB - Ensuring that people with an intellectual disability have normative levels of life satisfaction is increasingly considered a worthy goal for service providers. This review concerns the determinants of such satisfaction and embeds this literature within the Homeostatic Theory of Subjective Well-Being. This posits that life satisfaction is under considerable endogenous control and, as a consequence, does not normally vary in sympathy with changes in the external environment. This situation changes, however, if the environment is sufficiently aversive to defeat such homeostatic control. Under these conditions the circumstances of living correlate with life satisfaction as they wrest control away from the homeostatic system. One important implication is that the measure of life satisfaction may or may not be a sensitive indicator for changes in service provision, depending on the functional status of the homeostatic system. MRDD Research Reviews 7:99-104, 2001. (c) 2001 Wiley-Liss, Inc. PMID- 11389565 TI - Quality of life of adults with mental retardation/developmental disabilities who live with family. AB - In this paper, we review the literature related to the quality of life of adults with mental retardation/developmental disabilities (MR/DD) who live at home with their families. We examine the nature of the relationships between adults with MR/DD and their parents and siblings, the social worlds of adults with MR/DD, age related functional and health issues that affect their quality of life, the range of services and supports provided to them, and familial efforts to plan for their continued well-being when parental care is no longer viable. Individual characteristics associated with these dimensions and/or more compromised quality of life profiles are identified. The paper concludes with recommendations for expanded research on the quality of life of adults with MR/DD who live in the parental home, a topic which has received markedly less attention than the quality of life of adults who live in publicly supported residential settings. MRDD Research Reviews 7:105-114, 2001. PMID- 11389567 TI - Social interaction interventions for youth with severe disabilities should emphasize interdependence. AB - An emphasis on independence may be limiting the outcomes achieved by social interaction research. Given the lack of correlation between intellectual or adaptive functioning level and meaningful social outcomes for people with severe disabilities, a focus on increasing skills as a mechanism for increasing social outcomes seems misplaced (or, at least, inefficient). It is argued here that a more productive route for researchers to follow will be to emphasize interdependence. Implications of such a focus will change where interventions are implemented, curricular foci, and the types of instructional strategies used to improve the social life of youth with severe disabilities. MRDD Research Reviews 7:122-127, 2001. PMID- 11389566 TI - Living circumstances of children and adults with mental retardation or developmental disabilities in the United States, Canada, England and Wales, and Australia. AB - The purpose of this article was to collate evidence to describe where people with mental retardation or developmental disabilities live in five developed countries: United States, Canada, England, Wales, and Australia. Family homes are important dwelling places for people with mental retardation. They are the home of the great majority of children with mental retardation and a considerable proportion of adults with mental retardation. The likelihood of placement outside the family home increases with adulthood and progressively as people age. Adults with mental retardation live in a wide variety of settings, with formal residential provision frequently dependent on the arrangements that the authorities responsible for providing service support have chosen to make. There has been a considerable move away from accommodating people in large segregated and geographically isolated institutions in the countries considered. However, the current range of accommodation includes much with a distinctively different character to the homes where other citizens live. Many people still live in larger groups than would be ordinarily found in typical homes and this may necessitate departure from the architectural norm. In all of the countries considered, there has been a recent trend towards small community settings, compatible with typical housing architecture. This appears furthest advanced in the U.S. but is discernible elsewhere. Availability of residential services at a national level varies between 100 and 155 places per 100,000 total population. Regional variation within countries is even greater. In no case is the national availability considered adequate to meet the demand arising from changing need or expectations. MRDD Research Reviews 7:115-121, 2001. (c) 2001 Wiley-Liss, Inc. PMID- 11389568 TI - Social relationships in the workplace. AB - As more workers with disabilities are competitively employed, there are numerous opportunities to form social relationships with their co-workers. Close social relationships are associated with a number of positive outcomes (e.g., happiness, less stress) and should be actively pursued for those individuals desiring them. In this paper, we describe the types of social interactions displayed in work settings employing workers with and without disabilities. We also discuss some of the social problems experienced by some workers with disabilities. Finally, we describe two general categories of intervention strategies that have been used to impact social relationships: (a) strategies that involve changing the social behaviors of workers with disabilities, and (b) natural support strategies. The results from these strategies are discussed, and future areas of research are described. MRDD Research Reviews 7:128-133, 2001. PMID- 11389569 TI - Supporting positive sibling relationships during childhood. AB - This paper reviews the research literature focusing on the interpersonal relationships between siblings when one child has a disability. Descriptive findings are presented that compare and contrast sibling warmth and positivity, engagement, and conflict in sibling pairs with and without a child with a disability. The social roles assumed by siblings are examined, as are developmental changes in role relationships. Research on the development of the sibling relationship in the family context is reviewed, as are findings concerning the effects of parent differential attention on the quality of the sibling relationship. MRDD Research Reviews 7:134-142, 2001. PMID- 11389570 TI - Supporting communication in young children with developmental disabilities. AB - The behavior of parents, adult caregivers, and peers comprises the critical features of community support for the development of communication in young children with developmental disabilities. In a bio-ecological model of development, communication development is the result of the interactions of individuals with specific characteristics, in particular contexts over time. From the perspective of this model, foundational findings of intervention research to current views of communication development in children with developmental disabilities are summarized. The contributions of individual child characteristics to child-caregiver interactions that support language development are illustrated based on research with children who have autism, Williams syndrome, Down syndrome, and children who use augmentative communication systems. Parent-child interaction and the quality and quantity of parent talk are discussed as factors in children's language development. The effects of young children's delayed language on their interactions with peers, the contributions of peers to children's language learning and use, and the critical features of classroom settings that support child language development are reviewed. MRDD Research Reviews 7:143-150, 2001. PMID- 11389571 TI - Mechanical ventilation in children with severe asthma. AB - Hospital admissions for childhood asthma have increased during the past few decades. The aim of this study was to describe the need for mechanical ventilation for severe asthma exacerbation in children in Finland from 1976 to 1995. We reviewed medical records and collected data retrospectively from all 5 university hospitals in Finland, thus covering the entire population of about 5 million. The endpoints selected were the number of admissions and readmissions leading to mechanical ventilation, duration of stay in the hospital, and mortality. Moreover, asthma medications prescribed prior to admission and administered in the intensive care unit (ICU), as well as the etiology of the exacerbation associated with mechanical ventilation were examined. Mechanical ventilation was required in 66 ICU admissions (59 patients). This constituted approximately 10% of all 632 admissions for acute asthma to an ICU. The number of admissions decreased from 1976 to 1995: 41 admissions between 1976 and 1985 vs. 25 admissions during the next 10-year period. The mean age at admission to the ICU was 3.6 years, and 46% of the patients were boys. Prior to the index admission, 70% of the patients had used asthma medication such as oral bronchodilator (50%), inhaled bronchodilator (20%), theophylline (38%), inhaled glucocorticoid (18%), oral glucocorticoid (5%), and cromoglycate (7%). Respiratory infection was by far the most common cause of all the exacerbations (61%), followed by food allergy (8%) and gastroesophageal reflux (3%). In 28% of cases the cause of the severe asthma exacerbation could not be identified. In the mechanically ventilated patients readmissions occurred 38 times between 1976 and 1985 vs. 5 times between 1986 and 1995. Five of the patients who received mechanical ventilation died, and in 3 of these patients asthma was the event causing death. In conclusion, there has been decrease in the number of first and repeat ICU admission for asthma requiring mechanical ventilation between 1970 and 1995. This trend occurred despite a simultaneous 5% yearly increase in hospital admissions for childhood asthma during these 2 decades. PMID- 11389572 TI - Relationship between bronchial hyperresponsiveness and development of asthma in children with chronic cough. AB - To evaluate the relationship between bronchial hyperresponsiveness (BHR) and the development of asthma in children with chronic cough, we performed methacholine inhalation challenges and transcutaneous oxygen pressure (tcPO2) measurements in 92 children with chronic cough aged from 1-13 years (55 boys and 37 girls; mean, 5.3 years) and followed them for > or = 10 years. Forty-four age-matched children with asthma (24 males and 20 females; mean, 6.5 years) and 44 age-matched children without cough or asthma served as controls (18 males and 26 females; mean, 4.6 years). Consecutive doubling doses of methacholine were inhaled until a 10% decrease in tcPO2 from baseline was observed. The cumulative dose of methacholine at the inflection point of the tcPO2 record (Dmin-PO2) was considered to represent hyperresponsiveness to inhaled methacholine. After 10 years or more of follow-up, 60 of the 92 subjects with cough answered our questionnaire, and 27/60 had been diagnosed with asthma. There was a statistical difference in Dmin-PO2 between the children who presented with chronic cough originally and who developed asthma (asthma-developed group) and those who did not develop asthma (asthma-free group). There was no difference in the value of Dmin-PO2 between the asthma-developed group and the asthma group, or between the asthma-free group and the age-matched control group. Among the children with chronic cough, there was no difference in Dmin-PO2 between girls and boys, either in the asthma-developed group or in the asthma-group. We conclude that 45% of the children with a chronic cough in early life developed asthma, and that BHR in children with chronic cough during the childhood period is a strong risk factor for the development of asthma. PMID- 11389573 TI - Eosinophil cationic protein in infants with respiratory syncytial virus bronchiolitis: predictive value for subsequent development of persistent wheezing. AB - Infants with acute bronchiolitis during the first months of life are at increased risk of developing persistent wheezing and bronchial asthma later in life. The study of eosinophil cationic protein (ECP) suggests that eosinophil-related inflammatory mechanisms may play a role in respiratory syncytial virus (RSV) bronchiolitis. The aim of our study was to verify whether serum ECP (s-ECP) measurements are useful in predicting the development of persistent wheezing in children affected by RSV bronchiolitis during a 5 years follow-up period. Forty eight infants were enrolled prospectively (mean age: 153.5 days). All had a clinical and radiological diagnosis of acute bronchiolitis and confirmed RSV infection. Peripheral eosinophil counts, levels of s-ECP, and serum IgE concentrations were measured during bronchiolitis. Five years later the children were re-evaluated in regard to their respiratory symptoms (standardized questionnaires) and atopic status (specific IgE levels). We observed significantly higher s-ECP levels (P < 0.001) at enrollment in subjects who developed persistent wheezing compared to subjects who did not show late wheezing. Initial s-ECP values allowed significant and correct prediction of persistent wheezing (P < 0.001). The risk to develop respiratory symptoms was 9.73 higher for infants with s-ECP levels > or = 8 microg/L than for those with s ECP levels <8 microg/L (P < 0.0001). In conclusion, our study suggests that s-ECP levels in infants with bronchiolitis are useful in predicting the risk to develop wheezing in the subsequent 5 years. PMID- 11389574 TI - Functional residual capacity and passive compliance measurements after antenatal steroid therapy in preterm infants. AB - Studies in preterm animal models have shown that antenatal corticosteroids enhance lung maturation by improving a variety of physiologic variables, including lung volumes. Changes in lung volume of preterm infants treated with a full course of antenatal steroids have not been investigated. We hypothesized that a full course of antenatal steroids would significantly increase functional residual capacity (FRC) in treated vs. untreated preterm infants. The objective of our study was to compare FRC and respiratory mechanics in steroid treated vs. untreated preterm infants. FRC and passive respiratory mechanics were prospectively studied within 36 hr of life in 20 infants (25-34 weeks of gestation) who had received a full course of antenatal steroids and in 20 matched untreated preterm infants. FRC was measured with the nitrogen washout method, and respiratory mechanics with the single-breath occlusion technique. Preterm infants who received steroids (n = 20; mean birth weight = 1,230 g; gestational age = 28.8 weeks) had a significantly higher FRC (29.5 vs. 19.3 mL/kg; P < 0.001) than untreated infants (n = 20; birth weight = 1,202 g; gestational age = 28.5 weeks). Passive respiratory system compliance was also increased in treated vs. untreated infants (P < 0.05). In conclusion, FRC and passive respiratory system compliance were significantly improved in preterm infants (25-34 weeks gestation) treated with a full course of antenatal steroids, compared to matched untreated infants. Although this study was not randomized, it confirms that antenatal steroids have important effects on pulmonary function that may contribute to a decreased risk of respiratory distress syndrome in treated preterm infants. PMID- 11389575 TI - Bronchodilatory effects of salbutamol, ipratropium bromide, and their combination: double-blind, placebo-controlled crossover study in cystic fibrosis. AB - The efficacy of inhaled sympathomimetic and anticholinergic agents on airway obstruction in cystic fibrosis (CF) has been proven in several studies. However, studies comparing combined therapy with monotherapy led to divergent results, probably due to different study designs, different dosages, and the small numbers of patients investigated. Therefore, we wanted to answer the question which inhalation has the best short term effect: a sympathomimetic or an anticholinergic agent, or the combination of both. We investigated 17 patients with CF on 4 successive days in the morning, using pulmonary function testing before and 30 min after inhalation. Each patient received aerosolized salbutamol (SB, maximum dose (max.) 2.5 mg), ipratropium bromide (IB, max. 0.5 mg), the combination of both, or placebo (normal saline) in a randomized, double-blind crossover design. The mean forced expiratory volume in the first second improved significantly (adjusted P-value < 0.017) after each treatment compared to placebo. Analysis of variance showed that SB and combination therapy with SB and IB were superior to IB alone, without significant difference between SB and combination therapy. Response of a patient to combined therapy was usually associated with response to SB. Long-term efficacy and side effects of treatment with bronchodilators still remain to be investigated after this short term study. We conclude that in CF patients bronchodilator therapy with sympathomimetic agents is usually sufficient. Only in cases with proven additional benefit from inhalation by anticholinergics should combination therapy be recommended. PMID- 11389576 TI - Defining an exacerbation of pulmonary disease in cystic fibrosis. AB - Despite the importance of pulmonary exacerbations in CF in both clinical and research settings, both published evidence and consensus are lacking concerning the criteria used to define an exacerbation. The use of hospitalization as a surrogate measure presupposes uniformity among clinicians in diagnosis and treatment of exacerbations. Our aims were to evaluate consensus among clinicians about the variables considered helpful in diagnosing an exacerbation requiring treatment. A comprehensive list of symptoms, signs, and investigations used to define exacerbations was compiled from published trials. A written self administered questionnaire included the list in age-appropriate groups to survey opinion about the helpfulness of each item, and the estimated proportion of patients admitted within a month of diagnosis of an exacerbation. This was sent to all clinicians managing CF patients in Australia. There were replies from 59/91 clinicians (65%), 41/60 (68%) from those managing children and 18/31 (58%) from those managing adults. Responses of those managing children and adults differed for 7/32 variables (Mann-Whitney test, P < 0.05). Clinic grouping did not show greater consensus among responses of pediatricians (Kruskal-Wallis test, P = 0.362). Consensus, >74% or <26% of respondents rating a variable helpful/very helpful, was found in only 50% of variables listed. Estimated admission rate within a month of diagnosis was 61% (30-100%) for those managing adults and 48% %5-100%) for pediatricians. A lack of consensus was found among clinicians managing CF about the variables considered in diagnosing an exacerbation. The estimated proportion admitted within a month of diagnosis was very variable. This demonstrated inhomogeneity in approach to diagnosis and management of an exacerbation suggests a significant heterogeneity of clinical care. PMID- 11389577 TI - Transition programs in cystic fibrosis centers: perceptions of pediatric and adult program directors. AB - There is a growing population of adults with cystic fibrosis (CF) and a need for development of adult CF programs. Recommendations for transfer of patients to an adult program include a transition program. Our goal was to assess the current status of transition programs in US CF centers. In addition, we sought to determine the problems related to the transfer of patients to adult programs as perceived by CF center program directors. A survey was sent in 1998 to 110 pediatric and 44 adult program directors at CF centers approved by the Cystic Fibrosis Foundation (CFF), with a response rate of 65.5% and 72.7%, respectively: 22.2% of pediatric centers reported having a non-CFF-approved adult program, and 38.9% had no specific adult program. About one fifth of pediatric centers cited lack of an adult CF physician as an impediment to establishing an adult program. Age (82% of programs; mean, 18.5 years), but not marriage (17.1%) or pregnancy (24.8%), was used as a criterion for transfer. Criteria precluding transfer included patient/family resistance (51.4%), disease severity (50.5%), and developmental delay (46.7%). The concept of transfer is introduced to the patient and family at the time of diagnosis in a minority (14%) of programs. Over one half of the patients did not meet the adult team until the time of transfer. Pediatricians reported higher perceived parent, patient, pediatric staff, and adult staff concerns about transition issues than did adult program directors. We conclude that there is a lack of standardized programs for transfer of CF patients from a pediatric to an adult care setting, and that there are differences between pediatric and adult program directors' perceptions of concerns that CF patients, their families, and the medical teams have about transfer. These differences may impede the successful transition of patients into an adult program. PMID- 11389578 TI - Inspiratory flow reserve in boys with Duchenne muscular dystrophy. AB - Patients with advanced muscular dystrophy frequently develop ventilatory failure. Currently respiratory impairment usually is assessed by measuring vital capacity and the mouth pressure generated during a maximal inspiratory maneuver (PI,max), neither of which directly measures ventilatory capacity. We assessed inspiratory flow reserve in 26 boys [mean (SD) age 12.8 (3.8) years] with Duchenne muscular dystrophy (DMD) without ventilatory failure and in 28 normal boys [mean (SD) age 12.6 (1.9) years] by analyzing the ratio between the largest inspiratory flow during tidal breathing (V'I,max(t)) and during a forced vital capacity maneuver (V'I,max(FVC), (V'I,max(t)/V'I,maxFVC). We have compared this ratio with the forced vital capacity FVC and PI,max measured at functional residual capacity. Mean PI,max was -90(30)cmH2O, average 112% (range 57-179%) of predicted values in control boys and -31(11)cmH2O, average 40% predicted values in DMD boys (control vs DMD, P < 0.001). FVC was reduced in DMD boys [59(20)% predicted values vs 86(10)% predicted values in controls, P < 0.01]. Absolute V'I,max(FVC) was strongly related to FVC in both control and DMD boys; V'I,max(FVC) (expressed as FVC. s(-1)) was not related to PI,max in either group. The mean V'I,max(t)/V'I,max(FVC); ratio was higher in DMD 0.22 (0.08) than in controls 0.12 (0.03) (P < 0.001) indicating a reduction in inspiratory flow reserve in DMD. Inspiratory flow reserve was within the normal range in 8 of 19 DMD patients with PI,max less than 50% of predicted values. We conclude that measurement of inspiratory flow reserve (V'I,max(t)/V'I,maxFVC ratio) provides a simple and direct assessment of dynamic inspiratory muscle function which is not replicated by static measurement of PI,max or vital capacity and might be useful in assessment of respiratory impairment in boys with Duchenne muscular dystrophy. Follow-up studies are required to establish whether measures of inspiratory flow reserve are of clinical value in predicting subsequent ventilatory failure. PMID- 11389579 TI - Reduction of oscillatory pressure along the endotracheal tube is indicative for maximal respiratory compliance during high-frequency oscillatory ventilation: a mathematical model study. AB - We hypothesized that during high-frequency oscillatory ventilation (HFOV), a reduction of peak-to-peak oscillatory pressure along the endotracheal tube is maximal when respiratory system compliance is maximal. We made a mathematical model of the endotracheal tube and the respiratory system of a neonate suffering from idiopathic respiratory distress syndrome (IRDS). The model consisted of linear viscous and inertive elements, a non-linear endotracheal tube resistance, and a non-linear compliance allowing for alveolar recruitment and overdistention. Respiratory compliance was maximal at the transition between maximal recruitment and minimal overdistention. A new variable, the oscillatory pressure ratio (OPR), was defined as the ratio between peak-to-peak oscillatory pressures at the distal end and the proximal opening of the endotracheal tube, respectively. The respiratory variables of four patients were fed into the model, and the relationship between respiratory system compliance and OPR was determined. OPR decreased as compliance increased, except for very low compliances below where 0.08 mL. cm H2O(-1), and OPR increased with increasing compliance. The relationship between mean airway pressure P(aw) and OPR revealed that the minimal OPR (range, 0.37-0.78) and maximal respiratory compliance coincided at the same P(aw). However, the relationship did depend on oscillation frequency, applied oscillatory pressure, and endotracheal tube resistance, parameters that may change during clinical application of HFOV. When 81 permutations of nominal and extreme respiratory variables were used in the model, the minimum OPR (0.60 +/- 0.23) and maximum compliance coincided in all cases. These model experiments support our hypothesis. The results indicate that the OPR may be a useful index to optimize lung expansion, where lung recruitment is maximal and overdistention minimal. In vivo tests will be needed to reveal the feasibility and reliability of such an index for biomedical and clinical application. PMID- 11389580 TI - Anti-inflammatory effects of macrolides in lung disease. AB - During the past five decades there has been increasing interest in the potential anti-inflammatory effects of macrolide antibiotics. Low-dose macrolide therapy has dramatically increased survival in patients with diffuse panbronchiolitis, a disease with many similarities to cystic fibrosis (CF). This has led to further investigation into the potential use of macrolides in chronic lung diseases with an inflammatory component. This review summarizes the proposed anti-inflammatory mechanisms for this group of antibiotics, and examines the effect of macrolides on modulation of the inflammatory pathways, neutrophil function, bronchoconstriction, Pseudomonas biofilm, mucus rheology, bacterial adherence, and multidrug-resistant protein. It discusses the current status of clinical studies in children with CF, bronchiectasis, and asthma. While there are much in vitro data for different proposed anti-inflammatory effects, randomized controlled clinical trials in children with CF, bronchiectasis, and asthma are still just beginning. The benefits and potential side effects need to be determined before routine use can be advised. PMID- 11389581 TI - Cystic fibrosis in three children with bronchopulmonary dysplasia. AB - Cystic fibrosis (CF) and bronchopulmonary dysplasia (BPD) are two common causes of chronic lung disease in children. Patients with BPD or CF often have recurrent respiratory symptoms, failure to thrive, and/or metabolic alkalosis during infancy and childhood. Thus, recognizing the diagnosis of CF in an infant with BPD can be difficult. We present three infants with both BPD and CF. The infants shared a history of respiratory distress and prolonged oxygen requirements. All three also had difficulty gaining weight, even after pancreatic enzyme supplementation was instituted. Metabolic alkalosis was observed in two infants. Previous studies in children with CF suggest that early diagnosis may impact both lung health and nutritional status. A high index of suspicion is necessary for clinicians to identify these children early and intervene with appropriate therapy. PMID- 11389583 TI - [Medicine and public health]. PMID- 11389582 TI - Familial dysautonomia: a diagnostic dilemma. chronic lung disease with signs of an autoimmune disease. AB - We present an 11-year-old girl with sensory and autonomic neurological dysfunction, and respiratory insufficiency caused by recurrent aspiration. The diagnosis of familial dysautonomia (FD) was confirmed by a missing axonal flare to histamine, miosis in response to conjunctival methacholine and homozygous polymorphic linked markers DS58(18) and DS159(7) on chromosome 9. Ashkenazi Jewish descent could not be ascertained by history. A variety of positive tests for autoantibodies were initially interpreted as evidence for systemic lupus erythematosus vs. overlap syndrome with pulmonary, cerebral, skin, and ocular involvement. The diagnosis of FD was delayed because of the rarity of this disorder in Germany (second case reported). We discuss possible explanations for the misleading immunological findings, including interference by antibodies binding to milk proteins used as blocking reagents in enzyme-linked immunoassays and circulating immune-complexes due to chronic aspiration pneumonitis. PMID- 11389584 TI - Enthalpic and entropic contributions to the mutational changes in the reduction potential of azurin. AB - The changes in the reduction potential of Pseudomonas aeruginosa and Alcaligenes denitrificans azurins following point mutations and residue ionizations were factorized into the enthalpic and entropic contributions through variable temperature direct electrochemistry experiments. The effects on the reduction enthalpy due to changes in the first coordination sphere of the copper ion, as in the Met121Gln and Met121His variants of A. denitrificans azurin, insertion of a net charge and alteration in the solvation properties and electrostatic potential in proximity of the metal site, as in the Met44Lys and His35Leu variants of P. aeruginosa azurin, respectively, and proton uptake/release in wild-type and mutated species could invariably be accounted for on the basis of simple coordination chemistry and/or electrostatic considerations. The concomitant changes in reduction entropy were found in general to contribute to the E degrees ' variation to a lesser extent as compared to the enthalpy changes. However, their effects were by no means negligible and in some instances were found to heavily contribute to (or even become the main determinant of) the observed change in reduction potential. Several lines of evidence indicate that the entropic effects are notably influenced by reduction-induced solvent reorganization effects. In particular, protein reduction tends to be favored on entropic grounds with increasing exposure of the copper site to the solvent. Moreover, enthalpy-entropy compensation phenomena are invariably observed when residue mutation or pH-induced conformational changes modify the solvent accessibility of the metal site or alter the H-bonding network in the hydration shell of the molecule. Therefore, in these cases, caution must be used in making predictions of E degrees ' changes simply based on Coulombic or coordination chemistry arguments. PMID- 11389585 TI - Activation of class III ribonucleotide reductase from E. coli. The electron transfer from the iron-sulfur center to S-adenosylmethionine. AB - The anaerobic ribonucleotide reductase (ARR) from E. coli is the prototype for enzymes that use the combination of S-adenosylmethionine (AdoMet) and an iron sulfur center for generating catalytically essential free radicals. ARR is a homodimeric alpha2 protein which acquires a glycyl radical during anaerobic incubation with a [4Fe-4S]-containing activating enzyme (beta) and AdoMet under reducing conditions. Here we show that the EPR-active S = 1/2 reduced [4Fe-4S]+ cluster is competent for AdoMet reductive cleavage, yielding 1 equiv of methionine and almost 1 equiv of glycyl radical. These data support the proposal that the glycyl radical results from a one-electron oxidation of the reduced cluster by AdoMet. Reduced protein beta alone is also able to reduce AdoMet but only in the presence of DTT. However, in that case, 2 equiv of methionine per reduced cluster was formed. This unusual stoichiometry and combined EPR and Mossbauer spectroscopic analysis are used to tentatively propose that AdoMet reductive cleavage proceeds by an alternative mechanism involving catalytically active [3Fe-4S] intermediate clusters. PMID- 11389586 TI - Human apolipoprotein B gene intestinal control region. AB - Recently, we reported that a 315 bp enhancer, located over 55 kilobases (kb) upstream of the transcriptional start site of the human apolipoprotein B (apoB) gene, was sufficient to direct high-level expression of human apoB transgenes in mice. In this report, we expand our analysis of the distant apoB intestinal control region (ICR), by examining the function of segments in the vicinity of the 315 bp intestinal enhancer (315 IE). DNaseI hypersensitivity (DH) studies of a 4.8 kb segment from the ICR revealed three new DH sites, in addition to the previously described DH1 region present within the 315 IE. DH2 mapped to a 485 bp segment (485 IE) immediately upstream of the 315 IE that exhibited strong intestinal enhancer activity in transient transfection experiments with intestine derived CaCo-2 cells. Within the DH2 region, an HNF-4/ARP-1 binding site was demonstrated by gel retardation experiments. A 1.8 kb segment incorporating the 485 IE was capable of driving expression of human apoB transgenes in the intestines of mice. Additionally, a third component of the apoB ICR was found about 1.2 kb downstream of the 315 IE, within a 1031 bp segment (1031 IE) that also harbored two DH sites, DH3 and DH4. This segment did not display enhancer activity but was capable of driving transgene expression in the intestine. The three components of the ICR displayed a similar pattern of apoB mRNA expression along the horizontal axis of the intestine. The previously characterized in vivo liver-specific elements of the apoB gene, namely, the second intron enhancer and the 5' upstream liver enhancer, did not play a role in intestinal expression of apoB transgenes in mice. PMID- 11389587 TI - The 5' boundary of the human apolipoprotein B chromatin domain in intestinal cells. AB - The 5' boundary of the chromosomal domain of the human apolipoprotein B (apoB) gene in intestinal cells has been localized and characterized. It is composed of two kinds of boundary elements; the first, functional boundary is an insulator activity exhibited by a 1.8 kb DNA fragment located between -58 and -56 kb upstream of the human apoB promoter. In this region, an enhancer-blocking activity has been mapped to a CTCF binding site that is located upstream of two apoB intestinal enhancers (IEs), the 315 IE and the 485 IE. The CTCF site represents a boundary between two types of chromatin structure: an open, DNaseI sensitive region 3' of the CTCF site containing the intestinal regulatory elements and a closed, DNaseI-resistant region 5' of the CTCF site. The 1.8 kb fragment harboring the CTCF site also insulated mini-white transgenes against position effects in Drosophila melanogaster. The second, structural boundary is represented by a nuclear matrix attachment region (MAR), situated about 3 kb 5' of the CTCF site. This MAR may represent the 5' anchorage site for a chromosomal loop that functions to bring the intestinal regulatory elements closer to the apoB promoter. PMID- 11389588 TI - Intercalation of the (1R,2S,3R,4S)-N6-[1-(1,2,3,4-tetrahydro-2,3,4 trihydroxybenz[a]anthracenyl)]-2'-deoxyadenosyl adduct in the N-ras codon 61 sequence: DNA sequence effects. AB - The structure of the bay region (1R,2S,3R,4S)-N6-[1-(1,2,3,4-tetrahydro-2,3,4 trihydroxybenz[a]anthracenyl)]-2'-deoxyadenosyl adduct at X(7) of 5' d(CGGACAXGAAG)-3'.5'-d(CTTCTTGTCCG)-3', incorporating codons 60, 61 (underlined), and 62 of the human N-ras protooncogene, was determined by NMR. This was the bay region benz[a]anthracene RSRS (61,3) adduct. The BA moiety intercalated above the 5'-face of the modified base pair. NOE connectivities between imino protons were disrupted at T16 and T17. Large chemical shifts at the lesion site were consistent with ring current shielding arising from the BA moiety. A large chemical shift dispersion was observed for the BA aromatic protons. An increased rise of 8.17 A was observed between base pairs A6 x T17 and X7 x T(16). The PAH moiety stacked with the purine ring of A6, the 5'-neighbor nucleotide. This resulted in buckling of the 5'-neighbor A6 x T17 base pair, evidenced by exchange broadening for the T17 imino resonance. It also interrupted sequential NOE connectivities between nucleotides C5 and A6. The A6 deoxyribose ring showed an increased percentage of the C3'-endo conformation. This differed from the bay region BA RSRS (61,2) adduct, in which the lesion was located at position X6 [Li, Z., Mao, H., Kim, H.-Y., Tamura, P. J., Harris, C. M., Harris, T. M., and Stone, M. P. (1999) Biochemistry 38, 2969-2981], but was similar to the benzo[a]pyrene BP SRSR (61,3) adduct [Zegar I. S., Chary, P., Jabil, R. J., Tamura, P. J., Johansen, T. N., Lloyd, R. S., Harris, C. M., Harris, T. M., and Stone, M. P. (1998) Biochemistry 37, 16516-16528]. The altered sugar pseudorotation at A6 appears to be common to both bay region BA RSRS (61,3) and BP SRSR (61,3) adducts. It could not be discerned if the C3'-endo conformation at A6 in the BA RSRS (61,3) adduct altered base pairing geometry at X7 x T16, as compared to the C2'-endo conformation. The structural studies suggest that the mutational spectrum of this adduct may be more complex than that of the BA RSRS (61,2) adduct. PMID- 11389589 TI - Structure-function evaluation of ER alpha and beta interplay with SRC family coactivators. ER selective ligands. AB - Analysis of estrogen receptor alpha and beta interplay with other transcription factors is critical to the understanding of how small molecules, the cognate ligands for these receptors, selectively regulate the mode and amplitude of gene transcription by affecting receptor activity. To better understand the molecular mechanisms of selective action of estrogen receptor ligands, we characterized estrogen receptor alpha and beta (ER) interaction with the p160 family of coactivators. We also investigated how these interactions are affected by binding of specific ligands. We show that ER alpha and beta utilize different LXXLL motifs for their interaction with p160 family members. We found that significant differences exist between the affinity of the nuclear receptor interacting domain (NRID) and interaction of separate LXXLL motifs with ERs. This result indicates that a single LXXLL motif is unlikely to be sufficient for interaction with receptors, and that regions other than LXXLL motifs also participate in ER-p160 complex formation. We found that ER alpha and beta have strong affinity preferences for particular coactivators. These results suggest that ER-mediated transcription is not driven by a random mixture of ER-coactivator complexes. We also show that some ER ligands are functionally specific. We describe a ligand that binds to both receptors, but enhances only ER beta interaction with SRC1 and SRC3 while exhibiting little effect on the ER alpha interaction with these proteins. Finally, we provide data that suggest how genistein may selectively recruit coactivators when liganded to ERs. It enhances the interaction of ERs with SRC1 and SRC3, but demonstrates a minimal effect on receptor interaction with DRIP205 and CBP. PMID- 11389590 TI - Association of calnexin with wild type and mutant AVPR2 that causes nephrogenic diabetes insipidus. AB - Over 155 mutations within the V2 vasopressin receptor (AVPR2) gene are responsible for nephrogenic diabetes insipidus (NDI). The expression and subcellular distribution of four of these was investigated in transfected cells. These include a point mutation in the seventh transmembrane domain (S315R), a frameshift mutation in the third intracellular loop (804delG), and two nonsense mutations that code for AVPR2 truncated within the first cytoplasmic loop (W71X) and in the proximal portion of the carboxyl tail (R337X). RT-PCR revealed that mRNA was produced for all mutant receptor constructs. However, no receptor protein, as assessed by Western blot analysis, was detected for 804delG. The S315R was properly processed through the Golgi and targeted to the plasma membrane but lacked any detectable AVP binding or signaling. Thus, this mutation induces a conformational change that is compatible with endoplasmic reticulum (ER) export but dramatically affects hormone recognition. In contrast, the W71X and R337X AVPR2 were retained inside the cell as determined by immunofluorescence. Confocal microscopy revealed that they were both retained in the ER. To determine if calnexin could be involved, its interaction with the AVPR2 was assessed. Sequential coimmunoprecipitation demonstrated that calnexin associated with the precursor forms of both wild-type (WT) and mutant receptors in agreement with its general role in protein folding. Moreover, its association with the ER-retained R337X mutant was found to be longer than with the WT receptor suggesting that this molecular chaperone also plays a role in quality control and ER retention of misfolded G protein-coupled receptors. PMID- 11389591 TI - Solution structure of the Reps1 EH domain and characterization of its binding to NPF target sequences. AB - The recently described EH domain recognizes proteins containing Asn-Pro-Phe (NPF) sequences. Using nuclear magnetic resonance (NMR) data, we determined the solution structure of the EH domain from the Reps1 protein and characterized its binding to linear and cyclic peptides derived from a novel targeting protein. The structure calculation included 1143 distance restraints and 122 angle restraints and resulted in structures with a root-mean-square deviation of 0.40 +/- 0.05 A for backbone atoms of superimposed secondary structural elements. The structure comprises two helix-loop-helix motifs characteristic of EF-hand domains. Titration data with NPF-containing peptides showed evidence of intermediate exchange on the NMR chemical shift time scale, which required an analysis that includes curve fitting to obtain accurate equilibrium constants and dissociation rate constants. The cyclic and linear peptides bound with similar affinities (Kd = 65 +/- 17 and 46 +/- 14 microM, respectively) and to the same hydrophobic pocket formed between helices B and C. The cyclic peptide formed a complex that dissociated more slowly (k(off) = 440 +/- 110 s(-1)) than the linear peptide (k(off) = 1800 +/- 250 s(-1)), but had little change in affinity because of the slower rate of association of the cyclic peptide. In addition, we characterized binding to a peptide containing a DPF sequence (Kd = 0.5 +/- 0.2 mM). The characterization of binding between the Reps1 EH domain and its target proteins provides information about their role in endocytosis. PMID- 11389592 TI - Modeling of the outer vestibule and selectivity filter of the L-type Ca2+ channel. AB - Using the KcsA bacterial K+ channel crystal structure [Doyle, D. A., et al. (1998) Science 280, 69-74] and the model of the outer vestibule of the Na+ channel [Lipkind, G. M., and Fozzard, H. A. (2000) Biochemistry 39, 8161-8170] as structural templates, we propose a structural model of the outer vestibule and selectivity filter of the pore of the Ca2+ channel (alpha1C or Ca(v)1.2). The Ca2+ channel P loops were modeled by alpha-helix-turn-beta-strand motifs, with the glutamate residues of the EEEE motif located in the turns. P loops were docked in the extracellular part of the inverted teepee structure formed by S5 and S6 alpha-helices with backbone coordinates from the M1 and M2 helices of the KcsA crystal structure. This construction results in a conical outer vestibule that tapers to the selectivity filter at the bottom. The modeled selectivity ring forms a wide open pore ( approximately 6 A) in the absence of Ca2+. When Ca2+ is present ( approximately 1 microM), all four glutamate side chains move to the center and form a cage around the dehydrated Ca2+ ion, blocking the pore. In the millimolar concentration range, Ca2+ also interacts with two low-affinity sites located externally and internally, which were modeled by the same carboxylate groups of the selectivity filter. Calculation of the resulting electrostatic potentials show that the single Ca2+ ion is located in an electrostatic trap. Only when three Ca2+ ions are bound simultaneously in the high- and low-affinity sites of the selectivity filter is Ca2+ able to overcome electrostatic attraction, permitting Ca2+ flux. PMID- 11389593 TI - Crystal structure of ATP sulfurylase from Penicillium chrysogenum: insights into the allosteric regulation of sulfate assimilation. AB - ATP sulfurylase from Penicillium chrysogenum is an allosterically regulated enzyme composed of six identical 63.7 kDa subunits (573 residues). The C-terminal allosteric domain of each subunit is homologous to APS kinase. In the presence of APS, the enzyme crystallized in the orthorhombic space group (I222) with unit cell parameters of a = 135.7 A, b = 162.1 A, and c = 273.0 A. The X-ray structure at 2.8 A resolution established that the hexameric enzyme is a dimer of triads in the shape of an oblate ellipsoid 140 A diameter x 70 A. Each subunit is divided into a discreet N-terminal domain, a central catalytic domain, and a C-terminal allosteric domain. Two molecules of APS bound per subunit clearly identify the catalytic and allosteric domains. The sequence 197QXRN200 is largely responsible for anchoring the phosphosulfate group of APS at the active site of the catalytic domain. The specificity of the catalytic site for adenine nucleotides is established by specific hydrogen bonds to the protein main chain. APS was bound to the allosteric site through sequence-specific interactions with amino acid side chains that are conserved in true APS kinase. Within a given triad, the allosteric domain of one subunit interacts with the catalytic domain of another. There are also allosteric-allosteric, allosteric-N-terminal, and catalytic catalytic domain interactions across the triad interface. The overall interactions-each subunit with four others-provide stability to the hexamer as well as a way to propagate a concerted allosteric transition. The structure presented here is believed to be the R state. A solvent channel, 15-70 A wide exists along the 3-fold axis, but substrates have access to the catalytic site only from the external medium. On the other hand, a surface "trench" links each catalytic site in one triad with an allosteric site in the other triad. This trench may be a vestigial feature of a bifunctional ("PAPS synthetase") ancestor of fungal ATP sulfurylase. PMID- 11389594 TI - Mechanistic implications of methylglyoxal synthase complexed with phosphoglycolohydroxamic acid as observed by X-ray crystallography and NMR spectroscopy. AB - Methylglyoxal synthase (MGS) and triosephosphate isomerase (TIM) share neither sequence nor structural similarities, yet the reactions catalyzed by both enzymes are similar, in that both initially convert dihydroxyacetone phosphate to a cis enediolic intermediate. This enediolic intermediate is formed from the abstraction of the pro-S C3 proton of DHAP by Asp-71 of MGS or the pro-R C3 proton of DHAP by Glu-165 of TIM. MGS then catalyzes the elimination of phosphate from this enediolic intermediate to form the enol of methylglyoxal, while TIM catalyzes proton donation to C2 to form D-glyceraldehyde phosphate. A competitive inhibitor of TIM, phosphoglycolohydroxamic acid (PGH) is found to be a tight binding competitive inhibitor of MGS with a K(i) of 39 nM. PGH's high affinity for MGS may be due in part to a short, strong hydrogen bond (SSHB) from the NOH of PGH to the carboxylate of Asp-71. Evidence for this SSHB is found in X-ray, 1H NMR, and fractionation factor data. The X-ray structure of the MGS homohexamer complexed with PGH at 2.0 A resolution shows this distance to be 2.30-2.37 +/- 0.24 A. 1H NMR shows a PGH-dependent 18.1 ppm signal that is consistent with a hydrogen bond length of 2.49 +/- 0.02 A. The D/H fractionation factor (phi = 0.43 +/- 0.02) is consistent with a hydrogen bond length of 2.53 +/- 0.01 A. Further, 15N NMR suggests a significant partial positive charge on the nitrogen atom of bound PGH, which could strengthen hydrogen bond donation to Asp-71. Both His-98 and His-19 are uncharged in the MGS-PGH complex on the basis of the chemical shifts of their Cdelta and C(epsilon) protons. The crystal structure reveals that Asp-71, on the re face of PGH, and His-19, on the si face of PGH, both approach the NO group of the analogue, while His-98, in the plane of PGH, approaches the carbonyl oxygen of the analogue. The phosphate group of PGH accepts nine hydrogen bonds from seven residues and is tilted out of the imidate plane of PGH toward the re face. Asp-71 and phosphate are thus positioned to function as the base and leaving group, respectively, in a concerted suprafacial 1,4-elimination of phosphate from the enediolic intermediate in the second step of the MGS reaction. Combined, these data suggest that Asp-71 is the one base that initially abstracts the C3 pro-S proton from DHAP and subsequently the 3-OH proton from the enediolic intermediate. This mechanism is compared to an alternative TIM-like mechanism for MGS, and the relative merits of both mechanisms are discussed. PMID- 11389595 TI - Tryptic digestion of soybean lipoxygenase-1 generates a 60 kDa fragment with improved activity and membrane binding ability. AB - Lipoxygenases are key enzymes in the metabolism of unsaturated fatty acids. Soybean lipoxygenase-1 (LOX-1), a paradigm for lipoxygenases isolated from different sources, is composed of two domains: a approximately 30 kDa N-terminal domain and a approximately 60 kDa C-terminal domain. We used limited proteolysis and gel-filtration chromatography to generate and isolate a approximately 60 kDa fragment of LOX-1 ("mini-LOX"), produced by trypsin cleavage between lysine 277 and serine 278. Mini-LOX was subjected to N-terminal sequencing and to electrophoretic, chromatographic, and spectroscopic analysis. Mini-LOX was found to be more acidic and more hydrophobic than LOX-1, and with a higher content of alpha-helix. Kinetic analysis showed that mini-LOX dioxygenates linoleic acid with a catalytic efficiency approximately 3-fold higher than that of LOX-1 (33.3 x 10(6) and 10.9 x 10(6) M(-1) x s(-1), respectively), the activation energy of the reaction being 4.5 +/- 0.5 and 8.3 +/- 0.9 kJ x mol(-1) for mini-LOX and LOX 1, respectively. Substrate preference, tested with linoleic, alpha-linolenic, and arachidonic acids, and with linoleate methyl ester, was the same for LOX-1 and mini-LOX, and also identical was the regio- and stereospecificity of the products generated thereof, analyzed by reversed-phase and chiral high-performance liquid chromatography, and by gas chromatography/mass spectrometry. Mini-LOX was able to bind artificial vesicles with higher affinity than LOX-1, but the binding was less affected by calcium ions than was that of LOX-1. Taken together, these results suggest that the N-terminal domain of soybean lipoxygenase-1 might be a built-in inhibitor of catalytic activity and membrane binding ability of the enzyme, with a possible role in physio(patho)logical conditions. PMID- 11389596 TI - Pseudoreversion of the catalytic activity of Y14F by the additional substitution(s) of tyrosine with phenylalanine in the hydrogen bond network of delta 5-3-ketosteroid isomerase from Pseudomonas putida biotype B. AB - Delta5-3-ketosteroid isomerase (KSI) from Pseudomonas putida Biotype B catalyzes the allylic isomerization of Delta5-3-ketosteroids to their conjugated Delta4 isomers via a dienolate intermediate. Two electrophilic catalysts, Tyr-14 and Asp 99, are involved in a hydrogen bond network that comprises Asp-99 Odelta2...O of Wat504...Tyr-14 Oeta...Tyr-55 Oeta.Tyr-30 Oeta in the active site of P. putida KSI. Even though neither Tyr-30 nor Tyr-55 plays an essential role in catalysis by the KSI, the catalytic activity of Y14F could be increased ca. 26-51-fold by the additional Y30F and/or Y55F mutation in the hydrogen bond network. To identify the structural basis for the pseudoreversion in the KSI, crystal structures of Y14F and Y14F/Y30F/Y55F have been determined at 1.8 and 2.0 A resolution, respectively. Comparisons of the two structures near the catalytic center indicate that the hydrogen bond between Asp-99 Odelta2 and C3-O of the steroid, which is perturbed by the Y14F mutation, can be partially restored to that in the wild-type enzyme by the additional Y30F/Y55F mutations. The kinetic parameters of the tyrosine mutants with the additional D99N or D99L mutation also support the idea that Asp-99 contributes to catalysis more efficiently in Y14F/Y30F/Y55F than in Y14F. In contrast to the catalytic mechanism of Y14F, the C4 proton of the steroid substrate was found to be transferred to the C6 position in Y14F/Y30F/Y55F with little exchange of the substrate 4beta-proton with a solvent deuterium based on the reaction rate in D2O. Taken together, our findings strongly suggest that the improvement in the catalytic activity of Y14F by the additional Y30F/Y55F mutations is due to the changes in the structural integrity at the catalytic site and the resulting restoration of the proton-transfer mechanism in Y14F/Y30F/Y55F. PMID- 11389597 TI - Identification of the regulatory subunit of Arabidopsis thaliana acetohydroxyacid synthase and reconstitution with its catalytic subunit. AB - Acetohydroxyacid synthase (EC 4.1.3.18; AHAS) catalyzes the initial step in the formation of the branched-chain amino acids. The enzyme from most bacteria is composed of a catalytic subunit, and a smaller regulatory subunit that is required for full activity and for sensitivity to feedback regulation by valine. A similar arrangement was demonstrated recently for yeast AHAS, and a putative regulatory subunit of tobacco AHAS has also been reported. In this latter case, the enzyme reconstituted from its purified subunits remained insensitive to feedback inhibition, unlike the enzyme extracted from native plant sources. Here we have cloned, expressed in Escherichia coli, and purified the AHAS regulatory subunit of Arabidopsis thaliana. Combining the protein with the purified A. thaliana catalytic subunit results in an activity stimulation that is sensitive to inhibition by valine, leucine, and isoleucine. Moreover, there is a strong synergy between the effects of leucine and valine, which closely mimics the properties of the native enzyme. The regulatory subunit contains a sequence repeat of approximately 180 residues, and we suggest that one repeat binds leucine while the second binds valine or isoleucine. This proposal is supported by reconstitution studies of the individual repeats, which were also cloned, expressed, and purified. The structure and properties of the regulatory subunit are reminiscent of the regulatory domain of threonine deaminase (EC 4.2.1.16), and it is suggested that the two proteins are evolutionarily related. PMID- 11389598 TI - Ricin A-chain inhibitors resembling the oxacarbenium ion transition state. AB - Ricin toxin A-chain (RTA) is expressed by the castor bean plant and is among the most potent mammalian toxins. Upon activation in the cytosol, RTA depurinates a single adenine from position 4324 of rat 28S ribosomal RNA, causing inactivation of ribosomes by preventing the binding of elongation factors. Kinetic isotope effect studies have established that RTA operates via a D(N)*A(N) mechanism involving an oxacarbenium ion intermediate with bound adenine [Chen, X.-Y., Berti, P. J., and Schramm, V. L. (2000) J. Am. Chem. Soc. 122, 1609-1617]. On the basis of this information, stem-loop RNA molecules were chemically synthesized, incorporating structural features of the oxacarbenium ion-like transition state. A 10-base RNA stem-loop incorporating (1S)-1-(9-deazaadenin-9-yl)-1,4-dideoxy-1,4 imino-D-ribitol at the depurination site binds four times better (0.57 microM) than the 10-base RNA stem-loop with adenosine at the depurination site (2.2 microM). A 10-base RNA stem-loop with 1,2-dideoxyribitol [(2R,3S)-2 (hydroxymethyl)-3-hydroxytetrahydrofuran] at the depurination site binds with a Kd of 3.2 microM and tightens to 0.75 microM in the presence of 9-deazaadenine. A similar RNA stem-loop with 1,4-dideoxy-1,4-imino-D-ribitol at the depurination site binds with a K(d) of 1.3 microM and improves to 0.65 micro;M with 9 deazaadenine added. When (3S,4R)-4-hydroxy-3-(hydroxymethyl)pyrrolidine was incorporated at the depurination site of a 14-base RNA stem-loop, the Kd was 0.48 microM. Addition of 9-deazaadenine tightens the binding to 0.10 microM whereas added adenine increases the affinity to 12 nM. The results of this study are consistent with the unusual dissociative D(N)*A(N) mechanism determined for RTA. Knowledge of this intermediate has led to the design and synthesis of the highest affinity inhibitor reported for the catalytic site of RTA. PMID- 11389599 TI - Resonance Raman and EPR investigations of the D251N oxycytochrome P450cam/putidaredoxin complex. AB - We have performed resonance Raman and electron paramagnetic resonance (EPR) studies on the dioxygen bound state of the D251N mutant of cytochrome P450cam (oxy-P450cam) and its complex with reduced putidaredoxin (Pd). The D251N oxy P450cam/Pd complex has a perturbed proton delivery mechanism and shows a significantly red-shifted UV-visible spectrum as observed in Benson et al. [Benson, D. E., Suslick, K. S., and Sligar, S. G. (1997) Biochemistry 36, 5104 5107]. The red shift has been interpreted to indicate a major perturbation of the electronic structure of the oxy-heme complex. However, we find no evidence that electron transfer has occurred from Pd to the heme active site of D251N oxy P450cam. This suggests that both electron and proton transfer are perturbed by the D251N mutation and that these processes may be coupled. Three oxygen isotope sensitive Raman features are identified in the Pd complex, and occur at 1137, 536, and 399 cm(-1). These values are not significantly different from those for WT or D251N oxy-P450cam. However, a careful examination of the oxygen stretching feature near 1137 cm(-1) reveals the presence of three peaks at 1131, 1138, and 1146 cm(-1), which we attribute to the presence of conformational substates in oxy-P450cam. A significant change in the conformational substate population is observed for the D251N oxy-P450cam when the Pd complex is formed. We suggest that the conformational population redistribution of oxy-P450cam, along with the red shifted electronic spectra, reflects a structural equilibrium of the oxy-heme that is perturbed upon Pd binding. We propose that this structural perturbation is connected to the effector function of Pd and may involve changes in the electron donation properties of the thiolate ligand. PMID- 11389600 TI - Comparative electron paramagnetic resonance study of radical intermediates in turnip peroxidase isozymes. AB - The occurrence of isozymes in plant peroxidases is poorly understood. Turnip roots contain seven season-dependent isoperoxidases with distinct physicochemical properties. In the work presented here, multifrequency electron paramagnetic resonance spectroscopy has been used to characterize the Compound I intermediate obtained by the reaction of turnip isoperoxidases 1, 3, and 7 with hydrogen peroxide. The broad (2500 G) Compound I EPR spectrum of all three peroxidases was consistent with the formation of an exchange-coupled oxoferryl-porphyrinyl radical species. A dramatic pH dependence of the exchange interaction of the [Fe(IV)=O por(*+)] intermediate was observed for all three isoperoxidases and for a pH range of 4.5-7.7. This result provides substantial experimental evidence for previous proposals concerning the protein effect on the ferro- or antiferromagnetic character of the exchange coupling of Compound I based on model complexes. Turnip isoperoxidase 7 exhibited an unexpected pH effect related to the nature of the Compound I radical. At basic pH, a narrow radical species ( approximately 50 G) was formed together with the porphyrinyl radical. The g anisotropy of the narrow radical Delta(g) = 0.0046, obtained from the high-field (190 and 285 GHz) EPR spectrum, was that expected for tyrosyl radicals. The broad g(x) edge of the Tyr* spectrum centered at a low g(x) value (2.00660) strongly argues for a hydrogen-bonded tyrosyl radical in a heterogeneous microenvironment. The relationship between tyrosyl radical formation and the higher redox potential of turnip isozyme 7, as compared to that of isozyme 1, is discussed. PMID- 11389601 TI - Characterization of a half-apo derivative of peptidylglycine monooxygenase. Insight into the reactivity of each active site copper. AB - A derivative of peptidylglycine monooxygenase which lacks the CuH center has been prepared and characterized. This form of the enzyme is termed the half-apo protein. Copper-to-protein stoichiometric measurements establish that the protein binds only one of the two copper centers (CuM and CuH) found in the native enzyme. Confirmation that the methionine-containing CuM has been retained has been obtained from EXAFS experiments which show that the characteristic signature of the Cu-S(Met) interaction is preserved. The half-apo derivative binds 1 equiv of CO per copper with an IR frequency of 2092 cm(-1), and this monocarbonyl also displays the Cu-S(Met) interaction in its EXAFS spectrum. These results allow unambiguous assignment of the 2092 cm(-1) band as a CuM-CO species. Binding of CO in the presence of peptide substrate was also investigated. In the native enzyme, substrate induced binding of a second CO molecule with an IR frequency of 2062 cm(-1), tentatively assigned to a CO complex of the histidine-containing CuH site. Unexpectedly, this reactivity is also observed in the half-apo derivative, although the intensity distribution of the CO stretches now indicates that the copper has been partially transferred to a second site, believed to be CuH. The implications of this observation are discussed in terms of a possible additional peptide binding site close to the CuH center. PMID- 11389602 TI - Regulation of inducible nitric oxide synthase by self-generated NO. AB - A ferric heme-nitric oxide (NO) complex can build up in mouse inducible nitric oxide synthase (iNOS) during NO synthesis from L-arginine. We investigated its formation kinetics, effect on catalytic activity, dependence on solution NO concentration, and effect on enzyme oxygen response (apparent KmO2). Heme-NO complex formation was biphasic and was linked kinetically to an inhibition of electron flux and catalysis in iNOS. Experiments that utilized a superoxide generating system to scavenge NO showed that the magnitude of heme-NO complex formation directly depended on the NO concentration achieved in the reaction solution. However, a minor portion of heme-NO complex (20%) still formed during NO synthesis even when solution NO was completely scavenged. Formation of the intrinsic heme-NO complex, and the heme-NO complex related to buildup of solution NO, increased the apparent KmO2 of iNOS by 10- and 4-fold, respectively. Together, the data show heme-NO complex buildup in iNOS is due to both intrinsic NO binding and to equilibrium binding of solution NO, with the latter predominating when NO reaches high nanomolar to low micromolar concentrations. This behavior distinguishes iNOS from the other NOS isoforms and indicates a more complex regulation is possible for its activity and oxygen response in biologic settings. PMID- 11389603 TI - Role of the PR intermediate in the reaction of cytochrome c oxidase with O2. AB - The first discernible intermediate when fully reduced cytochrome c oxidase reacts with O2 is a dioxygen adduct (compound A) of the binuclear heme iron-copper center. The subsequent decay of compound A is associated with transfer of an electron from the low-spin heme a to this center. This reaction eventually produces the ferryl state (F) of this center, but whether an intermediate state may be observed between A and F has been the subject of some controversy. Here we show, using both optical and EPR spectroscopy, that such an intermediate (P(R)) indeed exists and that it exhibits spectroscopic properties quite distinct from F. The optical spectrum of P(R) is similar or identical to the spectrum of the P(M) intermediate that is formed after compound A when two-electron-reduced enzyme reacts with O2. An unusual EPR spectrum with features of a CuB(II) ion that interacts magnetically with a nearby paramagnet [cf. Hansson, O., Karlsson, B., Aasa, R., Vanngard, T., and Malmstrom, B.G (1982) EMBO J. 1, 1295-1297; Blair, D. F., Witt, S. N., and Chan, S. I. (1985) J. Am. Chem. Soc. 107, 7389 7399] can be uniquely assigned to the P(R) intermediate, not being found in either the P(M) or F intermediate. The binuclear center in the P(R) state may be assigned as having an Fe(a3)(IV)=O CuB(II) structure, as in both the P(M) and F states. The spectroscopic differences between these three intermediates are evaluated. The P(R) state has a key role as an initiator of proton translocation by the enzyme, and the thermodynamic and electrostatic bases for this are discussed. PMID- 11389604 TI - Identification of the proton pathway in bacterial reaction centers: cooperation between Asp-M17 and Asp-L210 facilitates proton transfer to the secondary quinone (QB). AB - The reaction center (RC) from Rhodobacter sphaeroides uses light energy to reduce and protonate a quinone molecule, Q(B) (the secondary quinone electron acceptor), to form quinol, Q(B)H2. Asp-L210 and Asp-M17 have been proposed to be components of the pathway for proton transfer [Axelrod, H. L., Abresch, E. C., Paddock, M. L., Okamura, M. Y., and Feher, G. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 1542 1547]. To test the importance of these residues for efficient proton transfer, the rates of the proton-coupled electron-transfer reaction k(AB)(2) (Q(A-*)Q(B-*) + H+ <==>Q(A-*)Q(B)H* --> Q(A)Q(B)H-) and its associated proton uptake were measured in native and mutant RCs, lacking one or both Asp residues. In the double mutant RCs, the k(AB)(2) reaction and its associated proton uptake were approximately 300-fold slower than in native RCs (pH 8). In contrast, single mutant RCs displayed reaction rates that were < or =3-fold slower than native (pH 8). In addition, the rate-limiting step of k(AB)(2) was changed from electron transfer (native and single mutants) to proton transfer (double mutant) as shown from the lack of a dependence of the observed rate on the driving force for electron transfer in the double mutant RCs compared to the native or single mutants. This implies that the rate of the proton-transfer step was reduced (> or =10(3)-fold) upon replacement of both Asp-L210 and Asp-M17 with Asn. Similar, but less drastic, differences were observed for k(AB)(1), which at pH > or =8 is coupled to the protonation of Glu-L212 [(Q(A-*)Q(B))-Glu- + H+ --> (Q(A)Q(B-*) GluH]. These results show that the pathway for proton transfer from solution to reduced Q(B) involves both Asp-L210 and Asp-M17, which provide parallel branches to the proton-transfer pathway and through their electrostatic interaction have a cooperative effect on the proton-transfer rate. A possible mechanism for the cooperativity is discussed. PMID- 11389605 TI - Solution and interface aggregation states of Crotalus atrox venom phospholipase A2 by two-photon excitation fluorescence correlation spectroscopy. AB - The dimeric Crotalus atrox venom PLA2 is part of the secreted phospholipase A2 (PLA2) enzyme family that interacts at the lipid-solution interface to hydrolyze the sn-2 acyl ester bond of phospholipids. We have employed fluorescence correlation spectroscopy (FCS) to study the monomer-dimer equilibrium of the C. atrox venom PLA2 in solution, in the presence of urea, and in the presence of monomeric and micellar n-dodecylphosphocholine (C12-PN), a phosphatidylcholine analogue. Dilution experiments show that PLA2 is an extremely tight dimer, Kd < or = 0.01 nM, in solution. Urea was introduced to weaken the subunit's association, and an estimate for the PLA(2) dimer dissociation constant in buffer was obtained by linear extrapolation. The derived dissociation constant was at least several orders of magnitude greater than that suggested from the dilution experiments, indicating a complex interaction between urea and the PLA2 dimer. FCS data indicate that the PLA2 dimer begins to dissociate at 10 mM C12-PN in 10 mM Ca2+ and at 5 mM C12-PN in 1 mM EDTA. The PLA2 tryptophan fluorescence displayed spectral shifts and intensity changes upon interacting with C12-PN. On the basis of the FCS and tryptophan fluorescence results, we postulate an intermediate state where the two monomers are in loose interaction within a protein-lipid comicelle. As the concentration of C12-PN was increased, complete dissociation of the dimer was observed, inferred from the doubling of the particle number, and the average diffusion constant decreased to approximately 60 microm2/s, consistent with PLA2 associated with a C12-PN micelle. The presence of Ca2+ makes the comicelle intermediate more stable, retarding the separation of the monomers in the micellar suspension. Our data clearly indicate that PLA2, though a strong dimer in the absence of lipids, is dissociated by micellar C12-PN and supports the monomer hypothesis for PLA2 action. PMID- 11389606 TI - A pH-induced dissociation of the dimeric form of a lysine 49-phospholipase A2 abolishes Ca2+-independent membrane damaging activity. AB - The hydrolysis of phospholipids by class II phospholipase A2 (PLA2) involves a Ca2+ ion cofactor bound to the Asp49 residue in the active site region. In the lysine 49 phospholipase A2 homologues (Lys49-PLA2), the Asp49 residue is substituted by Lys, and consequently the Lys49-PLA2s show no Ca2+ binding and no detectable phospholipid hydrolysis. Nevertheless, the Lys49-PLA2s demonstrate membrane damaging activity by an incompletely understood Ca2+-independent mechanism of action. Using a combination of steady-state and time-resolved fluorescence techniques, we have examined the effect of pH on the monomer-dimer equilibrium of bothropstoxin I (BthTX-I), a Lys49-PLA2 from the venom of Bothrops jararacussu which contains a single Trp77 residue located at the dimer interface. At pH 5.0, we observe a decreased quantum yield, a decreased rotational correlation time, and an increased bimolecular quenching rate constant with iodide. These results are consistent with a pH-induced dissociation of the BthTX I dimer, with the consequent exposure of the Trp77 residue to aqueous solvent. In the presence of liposomes, membrane damaging activity is observed only under conditions in which the dimeric form of the BthTX-I is favored. These results demonstrate that the dimeric form of the protein is essential for the initiation of the Ca2+-independent membrane damaging activity. PMID- 11389607 TI - Trajectory of nucleosomal linker DNA studied by fluorescence resonance energy transfer. AB - While the structure of the nucleosome core is known in atomic detail, the precise geometry of the DNA beyond the core particle is still unknown. We have used fluorescence resonance energy transfer (FRET) for determining the end-to-end distance of DNA fragments assembled with histones into nucleosomes. The DNA of a length of 150-220 bp was labeled with rhodamine-X on one end and fluorescein or Alexa 488 on the other. Assembling nucleosomes on these DNA fragments leads to a measurable energy transfer. The end-to-end distance computed from the FRET increases from 60 +/- 5 A at 150 bp to 75 +/- 5 A at 170 bp without measurable change above it. These distances are compatible with different geometries of the linker DNA, all having in common that no crossing can be observed up to 220 bp. Addition of H1 histone leads to an increase in energy transfer, indicating a compaction of the linker DNA toward the nucleosome. PMID- 11389608 TI - Quantitative analysis of the isolated GAAA tetraloop/receptor interaction in solution: a site-directed spin labeling study. AB - The GNRA (N: any nucleotide; R: purine) tetraloop/receptor interaction is believed to be one of the most frequently occurring tertiary interaction motifs in RNAs, but an isolated tetraloop/receptor complex has not been identified in solution. In the present work, site-directed spin labeling is applied to detect tetraloop/receptor complex formation and estimate the free energy of interaction. For this purpose, the GAAA tetraloop/receptor interaction was chosen as a model system. A method was developed to place nitroxide labels at specific backbone locations in an RNA hairpin containing the GAAA tetraloop. Formation of the tetraloop/receptor complex was monitored through changes in the rotational correlation time of the tetraloop and the attached nitroxide. Results show that a hairpin containing the GAAA tetraloop forms a complex with an RNA containing the 11-nucleotide GAAA tetraloop receptor motif with an apparent Kd that is strongly dependent on Mg2+. At 125 mM MgCl2, Kd = 0.40 +/- 0.05 mM. The corresponding standard free energy of complex formation is -4.6 kcal/mol, representing the energetics of the tetraloop/receptor interaction in the absence of other tertiary constraints. The experimental strategy presented here should have broad utility in quantifying weak interactions that would otherwise be undetectable, for both nucleic acids and nucleic acid-protein complexes. PMID- 11389609 TI - Phospholipid flippase activity of the reconstituted P-glycoprotein multidrug transporter. AB - The P-glycoprotein multidrug transporter acts as an ATP-powered efflux pump for a large variety of hydrophobic drugs, natural products, and peptides. The protein is proposed to interact with its substrates within the hydrophobic interior of the membrane. There is indirect evidence to suggest that P-glycoprotein can also transport, or "flip", short chain fluorescent lipids between leaflets of the membrane. In this study, we use a fluorescence quenching technique to directly show that P-glycoprotein reconstituted into proteoliposomes translocates a wide variety of NBD lipids from the outer to the inner leaflet of the bilayer. Flippase activity depended on ATP hydrolysis at the outer surface of the proteoliposome, and was inhibited by vanadate. P-Glycoprotein exhibited a broad specificity for phospholipids, and translocated phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, and sphingomyelin. Lipid derivatives that were flipped included molecules with long, short, unsaturated, and saturated acyl chains and species with the NBD group covalently linked to either acyl chains or the headgroup. The extent of lipid translocation from the outer to the inner leaflet in a 20 min period at 37 degrees C was directly estimated, and fell in the range of 0.36-1.83 nmol/mg of protein. Phospholipid flipping was inhibited in a concentration-dependent, saturable fashion by various substrates and modulators, including vinblastine, verapamil, and cyclosporin A, and the efficiency of inhibition correlated well with the affinity of binding to Pgp. Taken together, these results suggest that P-glycoprotein carries out both lipid translocation and drug transport by the same path. The transporter may be a generic flippase for hydrophobic molecules with the correct steric attributes that are present within the membrane interior. PMID- 11389610 TI - Characterization of the lysyl adducts of prostaglandin H-synthases that are derived from oxygenation of arachidonic acid. AB - These investigations characterize the covalent binding of reactive products of prostaglandin H-synthases (PGHSs) to the enzyme and to other molecules. The intermediate product of oxygenation of arachidonic acid by the PGHSs, prostaglandin (PG) H2, undergoes rearrangement to the highly reactive gamma-keto aldehydes, levuglandin (LG) E2 and D2. We previously have demonstrated that LGE2 reacts with the epsilon-amine of lysine to form both the lysyl-levuglandin Shiff base and the pyrrole-derived lysyl-levuglandin lactam adducts. We now demonstrate that these lysyl-levuglandin adducts are formed on the PGHSs following the oxygenation of arachidonic acid; after reduction of the putative Schiff base, proteolytic digestion of the enzyme, and isolation of the adducted amino acid residues, these adducts were identified by liquid chromatography-tandem mass spectrometry. The reactivity of the LGs is reflected by the finding that virtually all of the LG predicted to be formed from PGH2 can be accounted for as adducts of the PGH-synthase and that oxygenation of arachidonic acid by PGH synthases also leads to the formation of adducts of other proteins present in the reaction solution. The reactivity of the PGH-synthase adducts themselves is demonstrated by the formation of intermolecular cross-links. PMID- 11389611 TI - Protein conformation change of myoglobin upon ligand binding probed by ultraviolet resonance Raman spectroscopy. AB - Conformational change of myoglobin (Mb) accompanied by binding of a ligand was investigated with 244 nm excited ultraviolet resonance Raman Spectroscopy (UVRR). The UVRR spectra of native sperm whale (sw) and horse (h) Mbs and W7F and W14F swMb mutants for the deoxy and CO-bound states enabled us to reveal the UVRR spectra of Trp7, Trp14, and Tyr151 residues, separately. The difference spectra between the deoxy and CO-bound states reflected the environmental or structural changes of Trp and Tyr residues upon CO binding. The W3 band of Trp7 near the N terminus exhibited a change upon CO binding, while Trp14 did not. Tyr151 in the C terminus also exhibited a definite change upon CO binding, but Tyr103 and Tyr146 did not. The spectral change of Tyr residues was characterized through solvent effects of a model compound. The corresponding spectral differences between CO- and n-butyl isocyanide-bound forms were much smaller than those between the deoxy and CO-bound forms, suggesting that the conformation change in the C- and N terminal regions is induced by the proximal side of the heme through the movement of iron. Although the swinging up of His64 upon binding of a bulky ligand is noted by X-ray crystallographic analysis, UVRR spectra of His for the n-butyl isocyanide-bound form did not detect the exposure of His64 to solvent. PMID- 11389612 TI - Baccatin III induces assembly of purified tubulin into long microtubules. AB - Baccatin III is widely considered to be an inactive derivative of Taxol. We have reexamined its effect on in vitro assembly of tubulin under a variety of conditions. We found baccatin III to be active in all circumstances in which Taxol is active: it assembled GTP-tubulin, GDP-tubulin, and microtubule protein into normal microtubules and stabilized these polymers against cold-induced disassembly. The effect of baccatin III on in vitro microtubule assembly was quantitatively assessed through determination of critical concentrations, which can be used to obtain the apparent equilibrium constants for the addition of tubulin subunits to growing microtubules. The apparent equilibrium constants for the growth reaction for baccatin III-induced GTP-tubulin and GDP-tubulin assembly measured at 37 degrees C were 4.2-4.6-fold less than those measured for Taxol induced GTP-tubulin and GDP-tubulin assembly. These data indicate that the entire Taxol side chain contributes only about -1 kcal/mol to the apparent standard free energy of microtubule growth at 37 degrees C regardless of the nature of the E site nucleotide. These data also support the idea that the majority of the interactions between Taxol and tubulin that affect this equilibrium occur between the baccatin portion of the molecule and the binding site. We have also observed a structural difference in microtubules formed using baccatin III and Taxol. Baccatin III-induced microtubules were routinely much longer than those assembled by Taxol, even when very high concentrations of baccatin III were employed. One interpretation of these data is that baccatin III and Taxol differ in their abilities to nucleate GTP-tubulin. This difference in activity may have bearing on the large disparity in cytotoxicity of the two molecules. PMID- 11389613 TI - Thermodynamics of three-way multibranch loops in RNA. AB - RNA multibranch loops (junctions) are loops from which three or more helices exit. They are nearly ubiquitous in RNA secondary structures determined by comparative sequence analysis. In this study, systems in which two strands combine to form three-way junctions were used to measure the stabilities of RNA multibranch loops by UV optical melting and isothermal titration calorimetry (ITC). These data were used to calculate the free energy increment for initiation of a three-way junction on the basis of a nearest neighbor model for secondary structure stability. Imino proton NMR spectra were also measured for two systems and are consistent with the hypothesized helical structures. Incorporation of the experimental data into the mfold and RNA structure computer programs has contributed to an improvement in prediction of RNA secondary structure from sequence. PMID- 11389614 TI - On the mechanism of alpha-helix to beta-sheet transition in the recombinant prion protein. AB - It is believed that the critical event in the pathogenesis of transmissible spongiform encephalopathies is the conversion of the prion protein from an alpha helical form, PrP(C), to a beta-sheet-rich conformer, PrP(Sc). Recently, we have shown that incubation of the recombinant prion protein under mildly acidic conditions (pH 5 or below) in the presence of low concentrations of guanidine hydrochloride results in a transition to PrP(Sc)-like beta-sheet-rich oligomers that show fibrillar morphology and an increased resistance to proteinase K digestion [Swietnicki, W., Morillas, M, Chen, S., Gambetti, P., and Surewicz, W. K. (2000) Biochemistry 39, 424-431]. To gain insight into the mechanism of this transition, in the present study we have characterized the biophysical properties of the recombinant human prion protein (huPrP) at acidic pH in the presence of urea and salt. Urea alone induces unfolding of the protein but does not result in protein self-association or a conversion to beta-sheet structure. However, a time dependent transition to beta-sheet structure occurs upon addition of both urea and NaCl to huPrP, even at a sodium chloride concentration as low as 50 mM. This transition occurs concomitantly with oligomerization of the protein. At a given protein and sodium chloride concentration, the rate of monomeric alpha-helix to oligomeric beta-sheet transition is strongly dependent on the concentration of urea. Low and medium concentrations of the denaturant accelerate the reaction, whereas strongly unfolding conditions are not conducive to the conversion of huPrP into an oligomeric beta-sheet-rich structure. The present data strongly suggest that partially unfolded intermediates may be involved in the transition of the monomeric recombinant prion protein into the oligomeric scrapie-like form. PMID- 11389617 TI - Regiochemistry in the Pauson-Khand reaction: has a trans effect been overlooked? AB - Alkyne-dicobalt hexacarbonyl complexes have been studied by DFT to examine whether electronic differences in the acetylenic substituents could play a role in determining the regiochemical outcome in the Pauson-Khand reaction. It appears that in some instances the regiochemistry is, in fact, governed exclusively by the electronic nature of the acetylenic substituents through the discriminant loss of a carbon monoxide ligand. PMID- 11389616 TI - Aminoglycoside-nucleic acid interactions: remarkable stabilization of DNA and RNA triple helices by neomycin. AB - The stabilization of poly(dA).2poly(dT) triplex, a 22-base DNA triplex, and poly(rA).2poly(rU) triple helix by neomycin is reported. The melting temperatures, the association and dissociation kinetic parameters, and activation energies (E(on) and E(off)) for the poly(dA).2poly(dT) triplex in the presence of aminoglycosides and other triplex binding ligands were determined by UV thermal analysis. Our results indicate that: (i) neomycin stabilizes DNA triple helices, and the double helical structures composed of poly(dA).poly(dT) are virtually unaffected. (ii) Neomycin is the most active and triplex-selective stabilization agent among all aminoglycosides, previously studied minor groove binders, and polycations. Its selectivity (DeltaT(m3-->2) vs DeltaT(m2)(-->)(1)) exceeds most intercalating drugs that bind to triple helices. (iii) Neomycin selectively stabilizes DeltaT(m3)(-->)(2) for a mixed 22-base DNA triplex containing C and T bases in the pyrimidine strand. (iv) The rate constants of formation of triplex (k(on)) are significantly enhanced upon increasing molar ratios of neomycin, making triplex association rates closer to duplex association rates. (v) E(on) values become more negative upon increasing concentration of aminoglycosides (paromomycin and neomycin). E(off) values do not show any change for most aminoglycosides except neomycin. (vi) Aminoglycosides can effectively stabilize RNA [poly(rA).2poly(rU)] triplex, with neomycin[being one of the most active ligands discovered to date (second only to ellipticine). (vii) The stabilization effect of aminoglycosides on triple helices is parallel to their toxic behavior, suggesting a possible role of intramolecular triple helix (H-DNA) stabilization by the aminoglycosides. PMID- 11389618 TI - Molecular umbrella-assisted transport of glutathione across a phospholipid membrane. AB - A di-walled molecular umbrella (1a) has been synthesized by acylation of the terminal amino groups of spermidine with cholic acid, followed by condensation with bis(3-O-[N-1,2,3-benzotriazin-4(3H)-one]yl)-5,5'-dithiobis-2-nitrobenzoate (BDTNB), and displacement with glutathione (gamma-Glu-Cys-Gly, GSH). Replacement of the sterol hydroxyls with sulfate groups, prior to displacement with GSH, afforded a hexasulfate analogue 1b. Both conjugates have been found to enter large unilamellar vesicles (200 nm diameter, extrusion) of 1-palmitoyl-2-oleoyl sn-glycero-3-phosphocholine (POPC), and to react with entrapped GSH to form oxidized glutathione (GSSG). Evidence for vesicular entry has come from the formation of oxidized glutathione (GSSG) within the interior of the vesicle, the appearance of the thiol form of the umbrella (USH), and the absence of release of GSH into the external aqueous phase. Results that have been obtained from monolayer experiments, together with the fact that the heavily sulfated conjugate is able to cross the phospholipid bilayer, have yielded strong inferential evidence for an "umbrella-like" action of these molecules as they cross the lipid bilayer. PMID- 11389619 TI - Total synthesis of epothilone B, epothilone D, and cis- and trans-9,10 dehydroepothilone D. AB - The phosphonium salt 35, representing one of the two principal subunits of the epothilones, was prepared from propargyl alcohol via heptenone 22. A Wittig reaction of the phosphorane from 35 with aldehyde 33, obtained from aldol condensation of ketone 27 with aldehyde 28, afforded 37. Seco acid 42 derived from 37 underwent lactonization to give cis-9,10-dehydroepothilone D (43) which was selectively reduced with diimide to yield epothilone D (4) and, after epoxidation, epothilone B (2). An alternative route to epothilone D employed alkyne 39, obtained from 33, in a Castro-Stephens reaction with allylic bromide 34 to furnish enyne 40. The latter was semi-hydrogenated to provide 37. Alkyne 46, prepared from alcohol 45, was converted to trans-vinylstannane 47 which, in a Stille coupling with allylic chloride 50, gave 51. Seco acid 52 derived from 51 underwent lactonization to give trans-9,10-dehydroepothilone D (54). Bioassay data comparing the antiproliferative activity and tubulin polymerization of 43 and 54 with epothilone B (2), epothilone D (4), and paclitaxel (7) showed that the synthetic analogues were less potent than their natural counterparts, although both retain full antiproliferative activity against a paclitaxel resistant cell line. No significant difference in potency was noted between cis analogue 43 and its trans isomer 54. PMID- 11389620 TI - Nematic liquid crystals with a tetrafluoroethylene bridge in the mesogenic core structure. AB - A dramatic increase of the clearing temperatures of liquid crystals based on bis(cyclohexyl)ethane 1 by 50 to 70 K can be achieved by the perfluorination of the central ethylene link. Conformational analysis indicates that this effect is due to the increased rigidity of the mesogenic core structure and to the suppression of conformers with a bent shape. Materials based on bis(cylohexyl)tetrafluoroethane 2 might play a crucial role as materials for the next generation of active matrix LCDs with reduced power consumption. PMID- 11389621 TI - Total structure determination of apratoxin A, a potent novel cytotoxin from the marine cyanobacterium Lyngbya majuscula. AB - Apratoxin A (1), a potent cytotoxin with a novel skeleton, has been isolated from the marine cyanobacterium Lyngbya majuscula Harvey ex Gomont. This cyclodepsipeptide of mixed peptide-polyketide biogenesis bears a thiazoline ring flanked by polyketide portions, one of which possesses an unusual methylation pattern. Its gross structure has been elucidated by spectral analysis, including various 2D NMR techniques. The absolute configurations of the amino acid-derived units were determined by chiral HPLC analysis of hydrolysis products. The relative stereochemistry of the new dihydroxylated fatty acid unit, 3,7-dihydroxy 2,5,8,8-tetramethylnonanoic acid, was elucidated by successful application of the J-based configuration analysis originally developed for acyclic organic compounds using carbon-proton spin-coupling constants ((2,3)J(C,H)) and proton-proton spin coupling constants ((3)J(H,H)); its absolute stereochemistry was established by Mosher analysis. The conformation of 1 in solution was mimicked by molecular modeling, employing a combination of distance geometry and restrained molecular dynamics. Apratoxin A (1) possesses IC(50) values for in vitro cytotoxicity against human tumor cell lines ranging from 0.36 to 0.52 nM; however, it was only marginally active in vivo against a colon tumor and ineffective against a mammary tumor. PMID- 11389622 TI - A triangle-square equilibrium of metallosupramolecular assemblies based on pd(II) and pt(II) corners and diazadibenzoperylene bridging ligands. AB - Tetraaryloxy-substituted diazadibenzoperylene bridging ligands 1a,b were employed in transition metal-directed self-assembly with Pd(II) and Pt(II) phosphane triflates 2a,b which resulted in complex dynamic equilibria between molecular triangles 3a-d and molecular squares 4a-d in solution. Characterization of the equilibria and assignment of the metallacycles was accomplished by (1)H and (31)P[(1)H] NMR spectroscopy in combination with electrospray ionization Fourier transform ion cyclotron mass spectrometry (ESI-FTICR-MS). It was found that the equilibria depend on several factors, such as the metal ion (Pd(2+) or Pt(2+)), the solvent, and the steric demand of the phenoxy substituents of the diazadibenzoperylene ligands 1a,b. Introduction of bulky tert-butyl groups in 1b shifts the equilibrium significantly in the direction of the molecular squares. Molecular dynamics simulations of the triangle and square structures revealed critical steric effects and restricted conformational flexibilities of the phosphane and diazadibenzoperylene ligands that help explain the distinct dynamic behavior observed in variable-temperature NMR studies. Concentration-dependent UV/vis and fluorescence spectroscopy revealed the limited stability of the assemblies and confirmed the reversible nature of the dynamic equilibria. PMID- 11389623 TI - Fabrication of micro reaction cages with tailored properties. AB - Hollow polyelectrolyte capsules in micro- and submicrometer size were prepared. Their interior was functionalized by a "ship in bottle" synthesis of copolymers. While the monomers permeated the capsule wall easily, the formed polymers remained in the capsule cage. The physicochemical properties of the capsule interior such as ion strength, pH, light absorption, and fluorescence could be controlled independently from the surrounding solvent by means of the chemical nature of the captured polymer. In case of polyelectrolytes the osmotic pressure of the counterions led to a swelling of the capsules which can be important for micromechanics. The functionalization with light-sensitive materials allowed selective photoreactions inside the capsules. Synthesis of polyelectrolytes at high concentration resulted in an intertwining of the capsule wall with the polymer. The modified walls behaved like ion exchange membranes and showed selectivity toward adsorption and permeation of organic ions. The modified capsules offer many possibilities for novel applications as containers for controlled precipitation, as nanoreactors for catalyzed reactions, or as sensors. PMID- 11389624 TI - Mechanism of dissolution of sparingly soluble electrolytes. AB - Recent constant composition dissolution studies of sparingly soluble calcium phosphates have revealed an interesting and unusual behavior in that the rates decreased, eventually resulting in effective suppression, even though the solutions remained undersaturated. Contrary to traditional theories of dissolution, these experimental results indicated the importance of not only the particle size on the dissolution rate but also the participation of critical phenomena. In these theories, it is assumed that when the dissolution reactions are initiated, they continue spontaneously until all solid phase has disappeared. In terms of these mechanisms, there are no critical phenomena in the dissolution mechanism. Although the crystal size decreases during dissolution, when the reaction is controlled by polypitting (formation and growth of pits), the edge free energy increases at the very first stage due to the creation of pits and dissolution steps. The constant composition experimental results demonstrate the development of surface roughness as the dissolution steps are formed, implying an increase of the total edge length during the reactions. In an exactly analogous mechanism to crystal growth, the participation of critical conditions involving dissolution steps is a possibility. In contrast to crystal growth, dissolution is a process of size reduction and, when the particle size is sufficiently reduced, critical phenomena become important so that the influence of size must be taken into consideration. This paper proposes such a model for dissolution reactions, and although these unusual phenomena probably apply to all mineral phases, they are more evident for sparingly soluble electrolytes in which the critical conditions are attained much more readily. PMID- 11389625 TI - Sulfur K-edge X-ray absorption spectroscopy of 2Fe-2S ferredoxin: covalency of the oxidized and reduced 2Fe forms and comparison to model complexes. AB - Ligand K-edge X-ray absorption spectroscopy (XAS) provides a direct experimental probe of ligand-metal bonding. In previous studies, this method has been applied to mononuclear Fe-S and binuclear 2Fe-2S model compounds as well as to rubredoxins and the Rieske protein. These studies are now extended to the oxidized and reduced forms of ferredoxin I from spinach. Because of its high instability, the mixed-valence state was generated electrochemically in the protein matrix, and ligand K-edge absorption spectra were recorded using an XAS spectroelectrochemical cell. The experimental setup is described. The XAS edge data are analyzed to independently determine the covalencies of the iron-sulfide and -thiolate bonds. The results are compared with those obtained previously for the Rieske protein and for 2Fe-2S model compounds. It is found that the sulfide covalency is significantly lower in oxidized FdI compared to that of the oxidized model complex. This decrease is interpreted in terms of H bonding present in the protein, and its contribution to the reduction potential E degrees is estimated. Further, a significant increase in covalency for the Fe(III)-sulfide bond and a decrease of the Fe(II)-sulfide bond are observed in the reduced Fe(III)Fe(II) mixed-valence species compared to those of the Fe(III)Fe(III) homovalent site. This demonstrates that, upon reduction, the sulfide interactions with the ferrous site decrease, allowing greater charge donation to the remaining ferric center. That is the dominant change in electronic structure of the Fe(2)S(2)RS(4) center upon reduction and can contribute to the redox properties of this active site. PMID- 11389626 TI - Variable-temperature microcrystal X-ray diffraction studies of negative thermal expansion in the pure silica zeolite IFR. AB - Variable-temperature single-crystal X-ray diffraction using a synchrotron X-ray source has allowed the mechanism of negative thermal expansion in the pure silica zeolite IFR to be studied in greater detail than was previously possible over the temperature range 30-557 K. The results have allowed the changes in average atomic position with temperature to be measured and the structural features that are important in negative thermal expansion to be identified. The structure of zeolite IFR can be split into two regions: columns of fused rings that expand with temperature and the intercolumn regions, which tend to contract on heating. These competing changes combine to produce a material that contracts parallel to the crystallographic a and b axes and expands in the c-direction. Correlations between zeolite structure and thermal expansivity are also reported. PMID- 11389627 TI - The conformational origin of the barrier to the formation of neighboring group assistance in glycosylation reactions: a dynamical density functional theory study. AB - Static and dynamical Density Functional Theory studies of 2,6-di-O-acetyl-3,4-O isopropylidene-D-galactopyranosyl cation have shown that this cation can exist in two conformers characterized as (2)S(O) and B(2,5), respectively. The (2)S(O) conformer has the O-2 acyl group equatorial with the carbonyl syn to H-2 and is populated by monocyclic oxocarbenium ions. These conformational features are present in the structurally related glycosyl donor ethyl 2,6-di-O-benzoyl-3,4-O isopropylidene-beta-D-galactothiopyranoside as determined by X-ray diffraction studies. The B(2,5) conformer has O-2 axial and allows the carbonyl to rotate and close the five-membered ring to form a bicyclic dioxolenium ion. Constraints based on natural internal coordinates were implemented to study this conformational transition. In this way the barrier to interconversion has been determined to be 34 kJ mol(-)(1) with a transition state characterized as (O)S(2) and a pathway involving pseudorotation. Thus, for the first time the structures and energetics of the key ions postulated to be involved in neighboring group assisted glycosylation reactions have been determined. PMID- 11389628 TI - Chemistry of the diazeniumdiolates. 3. Photoreactivity. AB - We have found O(2)-substituted diazeniumdiolates, compounds of structure R(2)N N(O)=NOR' that are under development for various possible pharmaceutical uses, to be rather photosensitive. With R = ethyl and R' = methyl, benzyl, or 2 nitrobenzyl, the observed product distributions suggest that two primary pathways are operative. A minor pathway involves the extrusion of nitrous oxide (N(2)O) with simultaneous generation of R(2)N(*) and R'O(*), which may then form amines, aldehydes, and alcohols. The major reaction pathway is an interesting photochemical cleavage of the N=N bond to form a nitrosamine (R(2)NN=O) and an oxygen-substituted nitrene (R'ON). The intermediacy of the O-nitrene was inferred from the production of abundant oxime, via rearrangement of the O-nitrene to a C nitroso compound (R'ON --> O=NR'), and subsequent tautomerization to the more stable oxime. Involvement of the O-nitrene was confirmed by trapping with 2,3 dimethyl-2-butene to form the aziridine and with oxygen to generate the nitrate ester. 2-Nitro substitution on the benzyl derivative had surprisingly little effect on the reaction course. For each compound examined, minor amounts of nitric oxide (NO), presumably produced by secondary photolysis of the nitrosamine, were observed. Time-resolved infrared experiments provided additional support for the above reaction pathways and confirmed that the nitrosamine is a primary photoproduct. We have also found that the relative contributions of the reaction pathways can be altered in certain derivatives. For example, when R' = 2,4-dinitrophenyl, the contribution of the nitrosamine/O nitrene-forming pathway was diminished. Pharmacological implications of these results are discussed. PMID- 11389629 TI - Chemistry of the diazeniumdiolates. 2. Kinetics and mechanism of dissociation to nitric oxide in aqueous solution. AB - Diazeniumdiolate ions of structure R(2)N[N(O)NO](-) (1) are of pharmacological interest because they spontaneously generate the natural bioregulatory species, nitric oxide (NO), when dissolved in aqueous media. Here we report the kinetic details for four representative reactivity patterns: (a) straightforward dissociation of the otherwise unfunctionalized diethylamine derivative 2 (anion 1, where R = Et) to diethylamine and NO; (b) results for the zwitterionic piperazin-1-yl analogue 4, for which the protonation state of the neighboring basic amine site is an important determinant of dissociation rate; (c) data for 5, a diazeniumdiolate derived from the polyamine spermine, whose complex rate equation can include terms for a variety of medium effects; and (d) the outcome for triamine 6 (R = CH(2)CH(2)NH(3)(+)), the most stable structure 1 ion identified to date. All of these dissociations are acid-catalyzed, with equilibrium protonation of the substrate preceding release of NO. Specific rate constants and pK(a) values for 2-6 have been determined from pH/rate profiles. Additionally, a hypsochromic shift (from approximately 250 to approximately 230 nm) was observed on acidifying these ions, allowing determination of a separate pK(a) for each substrate. For 6, the pK(a) value obtained kinetically was 2-3 pK(a) units higher than the value obtained from the spectral shift. Comparison of the ultraviolet spectra for 6 at various pH values with those for O- and N alkylated diazeniumdiolates suggests that protonation at the R(2)N nitrogen initiates dissociation to NO at physiological pH, with a second protonation (at oxygen) accounting for both the spectral change and the enhanced dissociation rate at pH <4. Our results help to explain the previously noted variability in dissociation rate of 5, whose half-life we found to increase by an order of magnitude when its concentration was raised from near-zero to 1 mM, and provide mechanistic insight into the factors that govern dissociation rates among diazeniumdiolates of importance as pharmacologic progenitors of NO. PMID- 11389630 TI - Endo-exo and facial stereoselectivity in the Diels-Alder reactions of 3 substituted cyclopropenes with butadiene. AB - A computational examination of the four modes of addition in the Diels-Alder reactions of 3-substituted cyclopropene derivatives (substituents: BH(2), CH(3), SiH(3), NH(2), PH(2), OH, SH, F, and Cl) with butadiene have been carried out at the B3LYP/6-31++G(d)//HF/6-31++G(d) level. The degree of stabilization of these derivatives at the ground state correlates with the electronegativity of the substituent. This attenuation of reactivity and differences in steric interactions are the only factors needed to explain both the high facial selectivity and the differences in the endo-exo selectivity seen in these reactions. Furthermore, evidence is presented that indicates that stabilization by an interaction involving the syn C-3 hydrogen of cyclopropene and butadiene is small or irrelevant in controlling the endo-exo selectivity of the Diels-Alder reaction. PMID- 11389631 TI - First synthesis and structural determination of a monomeric, unsolvated lithium amide, LiNH(2). AB - Alkali metal amides typically aggregate in solution and the solid phase, and even in the gas phase. In addition, even in the few known monomeric structures, the coordination number of the alkali metal is raised by binding of Lewis-basic solvent molecules, with concomitant changes in structure. In contrast, the simplest lithium amide LiNH(2) has never been made in a monomeric form, even though its structure has been theoretically predicted several times. Here, the first experimental structural data for a monomeric, unsolvated lithium amide are determined using a combination of gas-phase synthesis and millimeter/submillimeter-wave spectroscopy. All data point to a planar structure for LiNH(2). The r(o) structure of LiNH(2) has a Li-N distance of 1.736(3) A, an N-H distance of 1.022(3) A, and a H-N-H angle of 106.9(1) degrees. These results are compared with theoretical predictions for LiNH(2), and experimental data for oligomeric, solid-phase samples, which could not resolve the question of whether LiNH(2) is planar or not. In addition, comparisons are made with revised gas phase and solid-phase data and calculated structures of NaNH(2). PMID- 11389632 TI - Protonation-induced paramagnetism. Structures and stabilities of six- and seven coordinate complexes of Os(II) in singlet and triplet states: a density functional study. AB - Li, Yeh, and Taube in 1993 (J. Am. Chem. Soc. 1993, 115, 10384) synthesized a number of complexes which can be formally regarded as protonated Os(II) species. Some of these were paramagnetic, in contrast to the diamagnetism of the closed shell 5d(6) Os(II) ions. This intriguing phenomenon is investigated theoretically using density functional theory. The geometries, stabilities, and electronic structures of a series of six- and seven-coordinate osmium complexes were studied in gas phase and aqueous solution using the B3P86 functional, in conjunction with the isodensity-polarized continuum model of solvation. The general formula for these complexes is [Os(NH(3))(4)H(L(1)(x)())(m)()(L(2)(y)())(n)()](()(x)()(+)(y)()(+3)+), where L(1) and L(2) = H(2)O, NH(3), CH(3)OH, CH(3)CN, Cl(-), and CN(-), which could be regarded as protonated Os(II) species or hydrides of Os(IV), although according to this work the osmium-hydrogen interaction is best described as a covalent Os(III)-H bond, in which the hydrogen is near-neutral. The ground states are generally found to be singlets, with low-lying triplet excited states. Solvation tends to favor the singlet states by as much as approximately 18 kcal mol(-)(1) in the 3+ ions, an effect which is proportional to the corresponding difference in molecular volumes. To have realistic estimates of the importance of spin-orbit coupling in these systems, the spin-orbit energy corrections were computed for triplet [Os(NH(3))(4)](2+), [Os(NH(3))(4)H](3+), and [Os(NH(3))(4)H(H(2)O)](3+), along with gas-phase Os and its ions as well as [Os(H(2)O)(6)](3+). The seven coordinate triplet-state complex [Os(NH(3))(5)H(CH(3)OH)](3+), which had been successfully isolated by Li, Yeh, and Taube, is predicted to be a stable six coordinate complex which strongly binds to a methanol molecule in the second coordination shell. The calculations further suggest that the singlet-triplet splitting would be very small, a few kilocalories per mole at most. The geometries and the electronic structures of the complexes are interpreted and rationalized in terms of Pauling's hybridization model in conjunction with conventional ligand field theory that effectively precludes the existence of true seven-coordinate triplet-state complexes of the above formula. PMID- 11389634 TI - Oxygen-carbon bond dissociation enthalpies of benzyl phenyl ethers and anisoles. An example of temperature dependent substituent effects. AB - For some time it has been assumed that the direction and magnitude of the effects of Y-substituents on the Z-X bond dissociation enthalpies (BDE's) in compounds of the general formula 4-YC(6)H(4)Z-X could be correlated with the polarity of the Z X bond undergoing homolysis. Recently we have shown by DFT calculations on 4 YC(6)H(4)CH(2)-X (X = H, F, Cl, Br) that the effects of Y on CH(2)-X BDE's are small and roughly equal for each X, despite large changes in C-X bond polarity. We then proposed that when Y have significant effects on Z-X BDE's it is due to their stabilization or destabilization of the radical. This proposal has been examined by studying 4-YC(6)H(4)O-X BDE's for X = H, CH(3), and CH(2)C(6)H(5) both by theory and experiment. The magnitudes of the effects of Y on O-X BDE's were quantified by Hammett type plots of DeltaBDE's vs sigma(+) (Y). Calculations reveal that changes in O-X BDE's induced by changing Y are large and essentially identical (rho(+) = 6.7-6.9 kcal mol(-)(1)) for these three classes of compounds. The calculated rho(+) values are close to those obtained experimentally for X = H at ca. 300 K and for X = CH(2)C(6)H(5) at ca. 550 K. However, early literature reports of the effects of Y on O-X BDE's for X = CH(3) with measurements made at ca. 1000 K gave rho(+) approximately 3 kcal mol(-)(1). We have confirmed some of these earlier, high-temperature O-CH(3) BDE's and propose that at 1000 K, conjugating groups such as -OCH(3) are essentially free rotors, and no longer lie mainly in the plane of the aromatic ring. As a consequence, the 298 K DFT calculated DeltaBDE for 4-OCH(3)-anisole of -6.1 kcal mol(-)(1) decreases to -3.8 kcal mol(-)(1) for free rotation, in agreement with the ca. 1000 K experimental value. In contrast, high-temperature O-CH(3) DeltaBDE's for three anisoles with strongly hindered substituent rotation are essentially identical to those that would be observed at ambient temperatures. We conclude that substituent effects measured at elevated temperatures may differ substantially from those appropriate for 298 K. PMID- 11389633 TI - Spectroscopy and reactivity of the type 1 copper site in Fet3p from Saccharomyces cerevisiae: correlation of structure with reactivity in the multicopper oxidases. AB - Fet3p is a multicopper oxidase recently isolated from the yeast, Saccharomyces cerevisiae. Fet3p is functionally homologous to ceruloplasmin (Cp) in that both are ferroxidases. However, by sequence homology Fet3p is more similar to fungal laccase, and both contain a type 1 Cu site that lacks the axial methionine ligand present in the functional type 1 sites of Cp. To determine the contribution of the electronic structure of the type 1 Cu site of Fet3p to the ferroxidase mechanism, we have examined the absorption, circular dichroism, magnetic circular dichroism, electron paramagnetic resonance, and resonance Raman spectra of wild type Fet3p and type 1 and type 2 Cu-depleted mutants. The spectroscopic features of the type 1 Cu site of Fet3p are nearly identical to those of fungal laccase, indicating a very similar three-coordinate geometry. We have also examined the reactivity of the type 1 Cu site by means of redox titrations and stopped-flow kinetics. From poised potential redox titrations, the E degrees of the type 1 Cu site is 427 mV, which is low for a three-coordinate type 1 Cu site. The kinetics of reduction of the type 1 Cu sites of four different multicopper oxidases with two different substrates were compared. The type 1 site of a plant laccase (Rhus vernicifera) is reduced moderately slowly by both Fe(II) and a bulky organic substrate, 1,4-hydroquinone (with 6 equiv of substrate, k(obs) = 0.029 and 0.013 s(-)(1), respectively). On the other hand, the type 1 site of a fungal laccase (Coprinus cinereus) is reduced very rapidly by both substrates (k(obs) > 23 s( )(1)). In contrast, both Fet3p and Cp are rapidly reduced by Fe(II) (k(obs) > 23 s(-)(1)), but only very slowly by 1,4-hydroquinone (10- and 100-fold more slowly than plant laccase, respectively). Semiclassical theory is used to analyze the origin of these differences in reactivity in terms of type 1 Cu site accessibility to specific substrates. PMID- 11389635 TI - Solvent effects on electrophilicity. AB - Continuum solvent effect on the electrophilicity index recently proposed by Parr and co-workers (Parr, R. G.; von Szentpaly, L.; Liu, S. J. Am. Chem. Soc. 1999, 121, 1922) is discussed in detail. Solvent effect is introduced using the self consistent isodensity polarized continuum model (SCI-PCM). A linear relationship is found between the change in electrophilicity index and the solvation energy as represented in the frame of the reaction field theory. The effect of a polarizable environment on the global electrophilicity is examined for a series of 18 well-known electrophiles presenting a wide diversity in structure and bonding properties. It is found that solvation enhances the electrophilicity power of neutral electrophilic ligands but attenuates this power in charged and ionic electrophiles. PMID- 11389636 TI - Thermal reactions of anti- and syn-dispiro[5.0.5.2]tetradeca-1,8-dienes: stereomutation and fragmentation to 3-methylcyclohexenes. Entropy-dictated product ratios from diradical intermediates? AB - A series of cyclobutanes substituted 1,2- by polyenes of increasing radical stabilizing power has been investigated to test the proposition that stabilization energies obtained independently from apposite, cis,trans geometric isomerizations can be successfully transferred to another system, in this paper, cyclobutanes. The first member of the series, 3-methylenecyclohexene (1), is photodimerized to anti- and syn-dispiro[5.0.5.2]tetradeca-1,8-dienes (anti-2 and syn-2), which undergo stereomutation (stereochemical interconversion) and cycloreversion (fragmentation) to 1 when heated in the range 72.1-118.2 degrees C: anti-2 --> syn-2, DeltaH() = 30.3 kcal mol(-)(1), DeltaS() = 0.2 cal mol(-)(1) K(-)(1); anti-2 --> 1, DeltaH() = 32.8 kcal mol(-)(1), DeltaS() = +8.0 cal mol( )(1) K(-)(1). Agreement with an enthalpy of activation predicted by assuming full allylic stabilization in a hypothetical diradical intermediate is good. An example of further activation by a radical-stabilizing group is manifested by the approximately 20 000-fold acceleration in rate shown by the system 1-phenyl-3 methylenecyclohexene (3) and anti- and syn-2,9-diphenyldispiro[5.0.5.2]tetradeca 1,8-dienes (anti-4 and syn-4), measured, however, only at 43.6 degrees C. In both systems 2 and 4, volumes of activation for stereochemical interconversion and cycloreversion have been determined and found to be essentially identical within experimental uncertainties, DeltaV() = +10.2 +/- 1.0 and +12.6 +/- 1.4 cm(3) mol( )(1), respectively (weighted means). These strongly positive values are consistent with the rate-determining step being the first bond-breaking, while the near identity of the volumes of activation argues against the indispensable second bond-breaking being a determining factor in fragmentation. These results are consistent with the theoretically based construct of Charles Doubleday for the paradigm, cyclobutane, in which the ratio between two channels of exit from a "generalized common biradical" is not controlled by enthalpy and entropy, as in the transition state model, but by entropy alone. PMID- 11389637 TI - An experimental and computational study on the reactivity and regioselectivity for the nitrosoarene ene reaction: comparison with triazolinedione and singlet oxygen. AB - The regioselectivities and the reactivities (relative rates) for the ene reaction of the enophile 4-nitronitrosobenzene (ArNO) with an extensive set of regiochemically defined acyclic and cyclic olefins have been determined. These experimental data establish that the ArNO enophile attacks the olefinic substrate along the novel skew trajectory, with preferred hydrogen abstraction at the corner (twix regioselectivity). This is in contrast to the isoelectronic species singlet oxygen ((1)O(2)), which abstracts at the higher substituted side of the double-bond (cis effect), and triazolindione (TAD), which undergoes the ene reaction at the more crowded end (gem effect). Ab initio computations (B3LYP/6 31+g) for the ene reaction of the ArNO with 2-methyl-2-butene reveal that the steric effects between the aryl group of the enophile and the substituents of the olefin dictate the skew trajectory. These computations identify the aziridine N oxide (AI) as a bona fide intermediate in this ene reaction, whose formation is usually rate-determining and, thus, irreversible along the skew trajectory (twix selectivity). The reversible generation of the AI becomes feasible when conformational constraints outweigh steric effects, as manifested by enhanced twin regioselectivity. PMID- 11389638 TI - Fabrication and characterization of metal-molecule-metal junctions by conducting probe atomic force microscopy. AB - Metal-molecule-metal junctions were fabricated by contacting Au-supported alkyl or benzyl thiol self-assembled monolayers (SAMs) with an Au-coated atomic force microscope (AFM) tip. The tip-SAM microcontact is approximately 15 nm(2), meaning the junction contains approximately 75 molecules. Current-voltage (I-V) characteristics of these junctions were probed as a function of SAM thickness and load applied to the microcontact. The measurements showed: (1) the I-V traces were linear over +/-0.3 V, (2) the junction resistance increased exponentially with alkyl chain length, (3) the junction resistance decreased with increasing load and showed two distinct power law scaling regimes, (4) resistances were a factor of 10 lower for junctions based on benzyl thiol SAMs compared to hexyl thiol SAMs having the same thickness, and (5) the junctions sustained fields up to 2 x 10(7) V/cm before breakdown. I-V characteristics determined for bilayer junctions involving alkane thiol-coated tips in contact with alkane thiol SAMs on Au also showed linear I-Vs over +/-0.3 V and the same exponential dependence on thickness. The I-V behavior and the exponential dependence of resistance on alkyl chain length are consistent with coherent, nonresonant electron tunneling across the SAM. The calculated conductance decay constant (beta) is 1.2 per methylene unit ( approximately 1.1 A(-)(1)) for both monolayer and bilayer junctions, in keeping with previous scanning tunneling microscope and electrochemical measurements of electron transfer through SAMs. These measurements show that conducting probe-AFM is a reliable method for fundamental studies of electron transfer through small numbers of molecules. The ability to vary the load on the microcontact is a unique characteristic of these junctions and opens opportunities for exploring electron transfer as a function of molecular deformation. PMID- 11389639 TI - The importance of the ene reaction for the C(2)-C(6) cyclization of enyne allenes. AB - The present study establishes the ene reaction as a competing reaction mechanism to the diradical mechanism for the thermal C(2)-C(6) cyclization of enyne-allenes which possess bulky substituents at the alkyne terminus. Both reaction routes are found to possess nearly equal free energies of activation. As shown by our computations, primary H/D isotope effects could be used for a definite decision about the mechanism. Concerning the regioselectivity of the cyclization reactions of enyne-allenes our study resolves a long-standing deviation between theoretical results and experimental findings. PMID- 11389640 TI - Potential energy surface for activation of methane by Pt(+): a combined guided ion beam and DFT study. AB - A guided-ion beam tandem mass spectrometer is used to study the reactions of Pt(+) with methane, PtCH(2)(+) with H(2) and D(2), and collision-induced dissociation of PtCH(4)(+) and PtCH(2)(+) with Xe. These studies experimentally probe the potential energy surface for the activation of methane by Pt(+). For the reaction of Pt(+) with methane, dehydrogenation to form PtCH(2)(+) + H(2) is exothermic, efficient, and the only process observed at low energies. PtH(+), formed in a simple C-H bond cleavage, dominates the product spectrum at high energies. The observation of a PtH(2)(+) product provides evidence that methane activation proceeds via a (H)(2)PtCH(2)(+) intermediate. Modeling of the endothermic reaction cross sections yields the 0 K bond dissociation energies in eV (kJ/mol) of D(0)(Pt(+)-H) = 2.81 +/- 0.05 (271 +/- 5), D(0)(Pt(+)-2H) = 6.00 +/- 0.12 (579 +/- 12), D(0)(Pt(+)-C) = 5.43 +/- 0.05 (524 +/- 5), D(0)(Pt(+)-CH) = 5.56 +/- 0.10 (536 +/- 10), and D(0)(Pt(+)-CH(3)) = 2.67 +/- 0.08 (258 +/- 8). D(0)(Pt(+)-CH(2)) = 4.80 +/- 0.03 eV (463 +/- 3 kJ/mol) is determined by measuring the forward and reverse reaction rates for Pt(+) + CH(4) right harpoon over left harpoon PtCH(2)(+) + H(2) at thermal energy. We find extensive hydrogen scrambling in the reaction of PtCH(2)(+) with D(2). Collision-induced dissociation (CID) of PtCH(4)(+), identified as the H-Pt(+)-CH(3) intermediate, with Xe reveals a bond energy of 1.77 +/- 0.08 eV (171 +/- 8 kJ/mol) relative to Pt(+) + CH(4). The experimental thermochemistry is favorably compared with density functional theory calculations (B3LYP using several basis sets), which also establish the electronic structures of these species and provide insight into the reaction mechanism. Results for the reaction of Pt(+) with methane are compared with those for the analogous palladium system and the differences in reactivity and mechanism are discussed. PMID- 11389641 TI - Direct evidence for preassociation preceding covalent binding in the reaction of cis-[Pt(NH3)2(H2O)2]2+ with surface immobilized oligonucleotides. PMID- 11389642 TI - Novel substituent and chelating effects in the Pd-catalyzed reaction of 2,3 allenols, aryl iodides, and amines. Highly regio- and stereoselective synthesis of 2-Amino-3-alken-1-ols or 4-Amino-2(E)-alken-1-ols. PMID- 11389643 TI - Simultaneous and stereoselective construction of planar and axial centers of chirality. PMID- 11389644 TI - Synthesis of hydrogels with extremely low surface friction. PMID- 11389645 TI - Novel phosphonium chloride-catalyzed dehydrohalogenative Si-C coupling reaction of alkyl halides with trichlorosilane. PMID- 11389646 TI - Supramolecular assembly of amide dendrons. PMID- 11389647 TI - Recognition and strand scission at junctions between single- and double-stranded DNA by a trinuclear copper complex. PMID- 11389648 TI - Formal synthesis of (+)-phorbol(1). PMID- 11389649 TI - DNA-block copolymer conjugates. PMID- 11389650 TI - Activation of an anilido ligand for nucleophilic aromatic substitution by an oxidizing Os(IV) center. PMID- 11389652 TI - Cyclometalated complexes: a new class of highly efficient photorefractive materials. PMID- 11389651 TI - The role of water bridges in directing the conformational preferences of 3-indole propionic acid and tryptamine. PMID- 11389653 TI - Palladium-catalyzed cross-coupling reaction of alkenyldimethyl(2-pyridyl)silanes with organic halides: complete switch from the carbometalation pathway to the transmetalation pathway. PMID- 11389654 TI - Asymmetric synthesis of beta-mercapto carboxylic acid derivatives by intramolecular sulfur transfer in N-enoyl oxazolidine-2-thiones promoted by Lewis acids. PMID- 11389655 TI - Proton-proton constraints in powdered solids from (1)H-(1)H-(1)H and (1)H-(1)H (13)C three-dimensional NMR chemical shift correlation spectroscopy. PMID- 11389657 TI - Coordination metallacycles of an achiral dendron self-assemble via metal-metal interaction to form luminescent superhelical fibers. PMID- 11389656 TI - Design and synthesis of the G-quadruplex-specific cleaving reagent perylene EDTA.iron(II). PMID- 11389658 TI - Gas-phase conformations and folding energetics of oligonucleotides: dTG- and dGT . PMID- 11389660 TI - A combinatorially-derived structural phase diagram for 42 zwitterionic geminis. PMID- 11389659 TI - Molecular recognition of titanium(IV) alkoxides by 2,6-bis(hydroxymethyl)-p cresol in the crystal engineering of hybrid organic-inorganic networks. PMID- 11389661 TI - A theoretical analysis of metal-molecule contacts. PMID- 11389662 TI - More information on the nature of stereotyped body-rocking. AB - Body-rocking was exposed to kinematic analysis in two studies. In the first study, amplitude was larger in the natural body-rocking of individuals with mental retardation than in the natural body-rocking of college students. Variability did not differ. In the second study, natural body-rocking of people with mental retardation was compared with their artificial body-rocking. Amplitude and variability were larger in the artificial condition. However, this result was not clear. We suggest that group and condition differences might have been a function of practice. PMID- 11389663 TI - Response restriction as a method to establish diurnal bladder control. AB - In this study, a new procedure to establish diurnal bladder control was evaluated. Eight children with severe and moderate levels of mental retardation participated. The procedure was based on the behavior theoretical concept of response restriction. Data were collected within a nonconcurrent multiple baseline design. Seven participants attained a level of bladder control in that they remained without urinary accidents between regular toileting times. Follow up measures revealed maintenance of these results. The results are discussed in terms of the advantages of this procedure as compared with existing procedures in this area, especially with respect to the omission of the restitutional overcorrection procedure. PMID- 11389664 TI - Receptive and expressive communication development of young males with fragile X syndrome. AB - We prospectively examined the developmental trajectories of receptive and expressive communication skills of 39 young males, 20 to 86 months of age, with fragile X syndrome. Eight showed features characteristic of autism. Children were tested one to three times using a standardized language test. They showed marked delays in language development, but substantial individual variability. Participants acquired expressive language skills more slowly than receptive language over time, gaining receptive language at about half the rate expected for typically developing children and expressive language at one third the rate. Both cognitive skills and autistic characteristics of the young males with fragile X syndrome related to receptive and expressive communication development, but neither predicted the discrepancies between expressive and receptive language acquisition over time. PMID- 11389665 TI - Prevalence of mental retardation and developmental disabilities: estimates from the 1994/1995 national health interview survey disability supplements. AB - In 1994 and 1995, the National Health Interview Survey included a Disability Supplement (NHIS-D) to collect extensive information about disabilities among individuals sampled as part of annual census-based household interview surveys. Here we describe the development and application of operational definitions of mental retardation and developmental disabilities to items in the NHIS-D to estimate prevalence. In our analyses, we estimate the prevalence of mental retardation in the noninstitutionalized population of the United States to be 7.8 people per thousand (.78%); of developmental disabilities, 11.3 people per thousand (1.13%); and the combined prevalence of mental retardation and/or developmental disabilities to be 14.9 per thousand (1.49%). Differences in prevalence estimates for mental retardation and developmental disabilities and among people of various ages are explored. PMID- 11389666 TI - Task-related social behavior in children with Down syndrome. AB - The specificity and stability of task-related social behaviors in children ages 5 to 12 was examined. Social behaviors during solvable and unsolvable puzzles were compared among children with Down syndrome, children with mental retardation, and typically developing children matched on mental age (MA). Compared to children without Down syndrome, those with the disorder looked to an adult and requested help more frequently. They also took longer to complete the tasks. These findings suggest that the overuse of social behaviors observed in young children with Down syndrome remains stable over the early school years and are specific to children with Down syndrome. PMID- 11389667 TI - Life course impacts of parenting a child with a disability. AB - We contrasted parents who had a child with a developmental disability, a serious mental health problem, and a normative comparison group with respect to parental attainment and well-being at mid-life. Data are from the Wisconsin Longitudinal Study, collected when the respondents were 18, 36, and 53 or 54, on average. Although similar at age 18, group patterns of attainment and well-being diverged thereafter. Parents of a child with a developmental disability had lower rates of employment, larger families, and lower rates of social participation but were similar to parents without a child with a disability in educational and marital status, physical health, and psychological well-being. Parents whose child had a serious mental health problem had normative patterns of educational and occupational attainment and marriage, but elevated levels of physical symptoms, depression, and alcohol symptoms at mid-life. PMID- 11389668 TI - Studies on lipase-affinity adsorption using response-surface analysis. AB - Lipases are widely distributed enzymes that can be obtained from animals, plants and micro-organisms. Coupling lipases with a wide range of substrates allows the opportunity for synthesis of optically pure pharmaceutical preparations, flavour compounds and other food additives. Affinity chromatography owes its power as a purification method to specific biological interactions. Response-surface analysis was chosen to study column efficiency. This method allows the understanding of interactions between variables with advantages over conventional methods, which involve changing one variable while fixing others at certain levels. The aim of this work was to study the influence of the ratio bed height/column diameter (L/D) and superficial velocity (V) on the column efficiency. The experimental design involved the two variables, L/D (2-10) and v (1-2 cm/min), at five levels. Lipase was obtained from Geotrichum sp. culture in a complex medium composed of 5% corn-steep liquor, 0.5% NH(4)NO(3) and 1% olive oil at 30 degrees C, with 1VVM (air volume/medium volume per min) aeration and 400 rev./min agitation. Maximum lipase activity was 19 units/ml after almost 9 h of fermentation. This lipase could potentially be used in esterification reactions to increase the content of gamma-linolenic acid and to produce bioaromas for food industries. The adsorption assays were carried out in a fixed bed column with an affinity adsorbent, which was obtained by reaction of a gel with oleic acid as ligand. Breakthrough curves were obtained for all experiments. It has been shown that the lower the values of both L/D and v, the higher the column efficiency (maximum 65.43%). Also, it was observed from the response surface that the efficiency reached a minimum at an L/D of around 8. PMID- 11389669 TI - Alginate as a macroaffinity ligand and an additive for enhanced activity and thermostability of lipases. AB - Lipases are being employed increasingly for the synthesis of drug intermediates and pharmaceutically important molecules as well as for the resolution of racemic mixtures for obtaining physiologically active enantiomers. Alginate was used as a macroaffinity ligand to purify lipases from Chromobacterium viscosum, porcine pancreas and wheatgerm by employing the technique of affinity precipitation. In all the cases, the purified preparation showed a single band on SDS/PAGE. The process gave adequate yields of 87, 75 and 62% in the case of Chromobacterium, porcine and wheatgerm lipases respectively. Alginate was also found to activate the enzymes; this effect was most dramatic in the case of wheatgerm lipase, for which a 4-5-fold activity increase was observed. Furthermore, alginate was also found to protect the enzyme against thermoinactivation. PMID- 11389670 TI - Rabies virus production in non-woven polyester fabric(NWPF) packed-bed reactors. AB - The production of rabies virus from baby hamster kidney-Ankara66 (BHK-An(66), HUKUK 99050302) monolayer cells was examinedin a packed-bed reactor containing non-woven polyester fabric (NWPF)discs. At first, growth characteristics of the cells were determined instatic culture. The suspension culture studies were realized in a 1litre spinner basket with NWPF support. BHK-An(66) cells(inoculation density, 5x10(4) cells.ml(-1)) were maintained in the reactor loaded with5 g.l(-1) carrier. The culture medium wasEagle's minimal essential medium supplemented with 10% (v/v)fetal bovine serum at 37 degrees C. During the culture, the mediumwas sampled daily to assess glucose and lactate concentrations. At theend of the 7 day culture period the cell density was found to be2.2x10(7) cells.ml(-1), and thereactor was inoculated with 5 ml [1.7x10(6) focus-forming units (ffu).ml(-1)] of the CVS 11(Challenge Virus Standard) strain of rabies virus. After a 72 hincubation period, the cultures were stained with fluorescein-conjugated anti-rabies globulin and were observed using a fluorescencemicroscope. Virus titres determined by the Spearman-Karber method were 2.2x10(5) ffu.ml(-1). In conclusion, NWPF packedreactors can be considered as a suitable system for the large-scaleproduction of rabies virus. PMID- 11389671 TI - Purification and conformational properties of a human interferon alpha2b produced in Escherichia coli. AB - Recombinant human interferon alpha2b was expressed intracellularly in Escherichia coli as insoluble aggregates using a new expression vector, and was purified to homogeneity using essentially two-step chromatographic procedures, i.e. immobilized metal-ion-affinity chromatography and reversed-phase HPLC. The established purification process is highly reproducible and leads to a total recovery of approx. 12% with a specific biological activity of higher than 1x10(8) i.u./mg of protein, which is comparable with the international requirement for interferon alpha2b. For purified protein we report conformational stability as a function of pH and temperature using differential scanning calorimetry and CD. Thermal unfolding as a function of pH showed only one endotherm at a temperature higher than 45 degrees C, and was reversible at pH 2 3.75 and irreversible at pH 4-10. At pH 7.0, the most stable condition, the conformational stability depends on protein concentration and ionic strength. The highly helical secondary structure is very conserved over the whole pH range studied, including at high temperatures. PMID- 11389672 TI - Studies on peptide amidase-catalysed C-terminal peptide amidation in organic media with respect to its substrate specificity. AB - Peptide amidase-catalysed amidations of the C- terminal carboxylic group of peptides were studied using model substrates of a large series of N(alpha) protected di-, tri-, tetra- and penta-peptides in the presence of NH4HCO3 as the ammonium source. The maximal yields of amide syntheses were achieved in a medium consisting of acetonitrile with 20-25 vol% of dimethylformamide and 3 vol% of water. Under these conditions, the substrate specificity of the enzyme was more restricted in the synthetic reaction than was found for the amide hydrolysis. Elongation of the peptide chain had a negative effect on enzymic amidation. Thus the direct amidation of N(alpha)-t-butoxycarbonyl-protected Leu-enkephalin resulted in a low yield of protected enkephalin amide. PMID- 11389673 TI - Structure and expression of an amylopullulanase gene from Bacillus stearothermophilus TS-23. AB - An amylopullulanase gene (apuTS) from Bacillus stearothermophilus TS-23 was cloned and characterized. apuTS consisted of an open reading frame of 6054 bp encoding a protein of 2018 amino acids with a calculated M(r) of 223811. The deduced amino acid sequence revealed four highly conserved regions that are common among amylolytic enzymes. In the C-terminal region, a six-amino-acid sequence (Pro-Gly-Ser-Gly-Thr-Thr) is repeated nine times. It shared the highest degree of homology with the amylopullulanase of Bacillus sp. XAL601. The enzyme also had moderate homology with amylopullulanases from thermophilic anaerobic bacteria. Low levels of homology were observed between the ApuTS of B. stearothermophilus TS-23 and amylopullulanases of Pyrococcus abyssi Orsay, P. furiosus and Bacillus sp. KSM1378. When the intact coding region of apuTS was expressed in Escherichia coli under the control of the lac promoter, the product was degenerate, as revealed by amylase activity staining after SDS/PAGE. The largest active polypeptide had an M(r) of about 220000, while the smallest one had an M(r) of about 105000. Upstream of the apuTS gene, a gene orfX was fortuitously cloned. The putative OrfX protein was weakly related to the myosin heavy chain. It was predicted to contain a central, 179-residue-long, coiled-coil domain. PMID- 11389674 TI - Immobilization of native and poly(ethylene glycol)-treated ('PEGylated') bovine serum amine oxidase into a biocompatible hydrogel. AB - Native bovine serum amine oxidase (BSAO) and poly(ethylene glycol) (PEG)-treated ('PEGylated') BSAO were immobilized into a hydrogel during its synthesis. The hydrogel was obtained by cross-linking of BSA with PEG di-nitrophenyl carbonates with a molecular mass of 10 kDa. Approx. 60% of the amino groups at the surface of BSAO were modified by monomethoxy-PEG with a molecular mass of 5 kDa when the reaction was carried out for 5 h in borate buffer, pH 9. The number of anchorage points of BSAO in the matrix, which was determined as minimal when PEGylated BSAO was used or maximal when native BSAO was used, did not influence the apparent K(m) and V(max) values of the different preparations. The apparent K(m) values of both forms of the enzyme were decreased due to preconcentration of benzylamine substrate by the negatively charged hydrogel. V(max) values were generally lower upon immobilization. We can therefore conclude that the hydrogel swelling has no significant effect on the enzyme's structure. The operational stability, evaluated in the presence of substrate, was generally increased upon enzyme immobilization into the hydrogel. Enzymic hydrogels were very stable during storage in solution at 4 degrees C, maintaining a high activity even after several weeks. The immobilization of both forms of BSAO did not improve their thermostability at 65 degrees C. The BSA-PEG hydrogel is a good matrix for immobilization of enzymes with therapeutic potential such as BSAO. PMID- 11389675 TI - Running-buffer composition influences DNA-protein and protein-protein complexes detected by electrophoretic mobility-shift assay (EMSA). AB - The gel-shift assay is a rapid, extremely sensitive, technically simple and widely used method for investigating nucleic acid-protein interaction based on the observation that binding of protein to DNA or RNA fragments usually leads to a reduction in the electrophoretic mobility of the fragment in non-denaturing gels. Here we report on the critical role of the running buffer and show that its importance ranks equally with other factors affecting complex formation and stability such as binding buffer, temperature, non-specific competitor or gel concentration and/or composition. We demonstrate differences in the binding patterns obtained with oligonucleotides containing binding sites for the ubiquitously expressed transcription factors Sp1 (stimulatory protein 1), NF-Y (nuclear factor Y) and USF (upstream stimulatory protein), which are dependent on the ionic strength of the running buffer used. Furthermore, we show the influence of glycine concentration on Sp1 binding using recombinant glutathione S transferase-Sp1 fusion protein expressed in Escherichia coli. PMID- 11389676 TI - What the structure of a calcium pump tells us about its mechanism. AB - The report of the crystal structure of the Ca(2+)-ATPase of skeletal muscle sarcoplasmic reticulum in its Ca(2+)-bound form [Toyoshima, Nakasako and Ogawa (2000) Nature (London) 405, 647-655] provides an opportunity to interpret much kinetic and mutagenic data on the ATPase in structural terms. There are no large channels leading from the cytoplasmic surface to the pair of high-affinity Ca(2+) binding sites within the transmembrane region. One possible access pathway involves the charged residues in transmembrane alpha-helix M1, with a Ca(2+) ion passing through the first site to reach the second site. The Ca(2+)-ATPase also contains a pair of binding sites for Ca(2+) that are exposed to the lumen. In the four-site model for transport, phosphorylation of the ATPase leads to transfer of the two bound Ca(2+) ions from the cytoplasmic to the lumenal pair of sites. In the alternating four-site model for transport, phosphorylation leads to release of the bound Ca(2+) ions directly from the cytoplasmic pair of sites, linked to closure of the pair of lumenal binding sites. The lumenal pair of sites could involve a cluster of conserved acidic residues in the loop between M1 and M2. Since there is no obvious pathway from the high-affinity sites to the lumenal surface of the membrane, transport of Ca(2+) ions must involve a significant change in the packing of the transmembrane alpha-helices. The link between the phosphorylation domain and the pair of high-affinity Ca(2+) binding sites is probably provided by two small helices, P1 and P2, in the phosphorylation domain, which contact the loop between transmembrane alpha-helices M6 and M7. PMID- 11389678 TI - Characterization of matrix metalloproteinase-26, a novel metalloproteinase widely expressed in cancer cells of epithelial origin. AB - Identification of expanding roles for matrix metalloproteinases (MMPs) in complex regulatory processes of tissue remodelling has stimulated the search for genes encoding proteinases with unique functions, regulation and expression patterns. By using a novel cloning strategy, we identified three previously unknown human MMPs, i.e. MMP-21, MMP-26 and MMP-28, in comprehensive gene libraries. The present study is focused on the gene and the protein of a novel MMP, MMP-26. Our findings show that MMP-26 is specifically expressed in cancer cells of epithelial origin, including carcinomas of lung, prostate and breast. Several unique structural and regulatory features, including an unusual 'cysteine-switch' motif, discriminate broad-spectrum MMP-26 from most other MMPs. MMP-26 efficiently cleaves fibrinogen and extracellular matrix proteins, including fibronectin, vitronectin and denatured collagen. Protein sequence, minimal modular domain structure, exon-intron mapping and computer modelling demonstrate similarity between MMP-26 and MMP-7 (matrilysin). However, substrate specificity and transcriptional regulation, as well as the functional role of MMP-26 and MMP-7 in cancer, are likely to be distinct. Despite these differences, matrilysin-2 may be a suitable trivial name for MMP-26. Our observations suggest an important specific function for MMP-26 in tumour progression and angiogenesis, and confirm and extend the recent findings of other authors [Park, Ni, Gerkema, Liu, Belozerov and Sang (2000) J. Biol. Chem. 275, 20540--20544; Uria and Lopez-Otin (2000) Cancer Res. 60, 4745--4751; de Coignac, Elson, Delneste, Magistrelli, Jeannin, Aubry, Berthier, Schmitt, Bonnefoy and Gauchat (2000) Eur. J. Biochem. 267, 3323--3329]. PMID- 11389679 TI - Role of the System L permease LAT1 in amino acid and iodothyronine transport in placenta. AB - The feto-placental unit relies on a maternal supply of indispensable amino acids and iodothyronines for early development and normal growth. We examined the role of the System L transporter in placental uptake of these substances, using the human placental choriocarcinoma cell line BeWo as a model experimental system. BeWo cells express both heavy (4F2hc) and light (LAT1, LAT2) chains of the System L holotransporter. Saturable transport of both L-[(3)H]tryptophan and [(125)I]tri iodo-L-thyronine in BeWo cells includes components sensitive to inhibition by the System-L-specific substrate 2-endoamino-bicycloheptane-2-carboxylic acid; kinetic properties of these components indicate that the 4F2hc-LAT1 transporter isoform is likely to predominate for the carriage of both substances at physiologically relevant concentrations. Both 4F2hc and LAT1 proteins are also expressed in human placental membranes and LAT1 at least is localized largely to the syncytiotrophoblast layer of the term human placenta. The 4F2hc-LAT1 transporter might therefore serve a vital role in supplying the developing fetus and the placenta with both thyroid hormones and indispensable amino acids from the maternal circulation. PMID- 11389677 TI - Structural similarities of Na,K-ATPase and SERCA, the Ca(2+)-ATPase of the sarcoplasmic reticulum. AB - The crystal structure of SERCA1a (skeletal-muscle sarcoplasmic reticulum/endoplasmic-reticulum Ca(2+)-ATPase) has recently been determined at 2.6 A (note 1 A = 0.1 nm) resolution [Toyoshima, Nakasako, Nomura and Ogawa (2000) Nature (London) 405, 647-655]. Other P-type ATPases are thought to share key features of the ATP hydrolysis site and a central core of transmembrane helices. Outside of these most-conserved segments, structural similarities are less certain, and predicted transmembrane topology differs between subclasses. In the present review the homologous regions of several representative P-type ATPases are aligned with the SERCA sequence and mapped on to the SERCA structure for comparison. Homology between SERCA and the Na,K-ATPase is more extensive than with any other ATPase, even PMCA, the Ca(2+)-ATPase of plasma membrane. Structural features of the Na,K-ATPase are projected on to the Ca(2+)-ATPase crystal structure to assess the likelihood that they share the same fold. Homology extends through all ten transmembrane spans, and most insertions and deletions are predicted to be at the surface. The locations of specific residues are examined, such as proteolytic cleavage sites, intramolecular cross-linking sites, and the binding sites of certain other proteins. On the whole, the similarity supports a shared fold, with some particular exceptions. PMID- 11389680 TI - Endosomal localization of phospholipase D 1a and 1b is defined by the C-termini of the proteins, and is independent of activity. AB - The factors regulating the activity of cellular phospholipase D (PLD) have been well characterized; however, the cellular distribution of specific PLD isoforms and the factors defining localization are less clear. Two specific PLD1 isoforms, PLD1a and PLD1b, are shown in the present study to be localized in endosomal compartments with early endosomal autoantigen 1, internalizing epidermal growth factor receptor (ErbB1) and lysobisphosphatidic acid. Novel C-terminal splice variants of PLD1, PLD1a2 and PLD1b2, do not exhibit this endosomal localization. Studies using catalytically inactive and C-terminal deletion mutants of the four PLD1 isoforms led to the conclusion that the C-terminus plays an important part in the catalytic activity of PLD1, but that the endosomal localization of PLD1a and PLD1b is defined by the C-terminus and not catalytic activity. PMID- 11389681 TI - A role of tensin in skeletal-muscle regeneration. AB - Regeneration of skeletal muscle requires the activation, proliferation, differentiation and fusion of satellite cells to generate new muscle fibres. This study was designed to determine the role of tensin in this process. Cardiotoxin was used to induce regeneration in the anterior tibial muscles of tensin-knockout and wild-type mice. From histological analysis, we found that the regeneration process lasted longer in knockout than in wild-type mice. To investigate the mechanism involved in this delay, we examined each regeneration step in animals and cultured primary cells. We found fewer proliferating myogenic cells identified by bromodeoxyuridine and desmin double labelling in knockout mice on the first 2 days after injury. Expression of myosin, paxillin, dystrophin and dystrophin-associated proteins were delayed in knockout mice. Withdrawal from the cell cycle was less efficient in isolated knockout myoblasts, and the fusion capacity was reduced in these cells as well. These defects in regeneration most likely contributed to the 9-fold increase of centrally nucleated fibres occurring in the non-injected knockout mice. Our results demonstrated clearly that tensin plays a role in skeletal-muscle regeneration. PMID- 11389682 TI - Reducible cationic lipids for gene transfer. AB - One of the main challenges of gene therapy remains the increase of gene delivery into eukaryotic cells. We tested whether intracellular DNA release, an essential step for gene transfer, could be facilitated by using reducible cationic DNA delivery vectors. For this purpose, plasmid DNA was complexed with cationic lipids bearing a disulphide bond. This reduction-sensitive linker is expected to be reduced and cleaved in the reducing milieu of the cytoplasm, thus potentially improving DNA release and consequently transfection. The DNA--disulphide-lipid complexation was monitored by ethidium bromide exclusion, and the size of complexes was determined by dynamic light scattering. It was found that the reduction kinetics of disulphide groups in DNA--lipid complexes depended on the position of the disulphide linker within the lipid molecule. Furthermore, the internal structure of DNA--lipid particles was examined by small-angle X-ray scattering before and after lipid reduction. DNA release from lipid complexes was observed after the reduction of disulphide bonds of several lipids. Cell transfection experiments suggested that complexes formed with selected reducible lipids resulted in up to 1000-fold higher reporter-gene activity, when compared with their analogues without disulphide bonds. In conclusion, reduction-sensitive groups introduced into cationic lipid backbones potentially allow enhanced DNA release from DNA--lipid complexes after intracellular reduction and represent a tool for improved vectorization. PMID- 11389683 TI - Fungal trehalose phosphorylase: kinetic mechanism, pH-dependence of the reaction and some structural properties of the enzyme from Schizophyllum commune. AB - Initial-velocity measurements for the phospholysis and synthesis of alpha,alpha trehalose catalysed by trehalose phosphorylase from Schizophyllum commune and product and dead-end inhibitor studies show that this enzyme has an ordered Bi Bi kinetic mechanism, in which phosphate binds before alpha,alpha-trehalose, and alpha-D-glucose is released before alpha-D-glucose 1-phosphate. The free-energy profile for the enzymic reaction at physiological reactant concentrations displays its largest barriers for steps involved in reverse glucosyl transfer to D-glucose, and reveals the direction of phospholysis to be favoured thermodynamically. The pH dependence of kinetic parameters for all substrates and the dissociation constant of D-glucal, a competitive dead-end inhibitor against D glucose (K(i)=0.3 mM at pH 6.6 and 30 degrees C), were determined. Maximum velocities and catalytic efficiencies for the forward and reverse reactions decrease at high and low pH, giving apparent pK values of 7.2--7.8 and 5.5--6.0 for two groups whose correct protonation state is required for catalysis. The pH dependences of k(cat)/K are interpreted in terms of monoanionic phosphate and alpha-D-glucose 1-phosphate being the substrates, and of the pK value seen at high pH corresponding to the phosphate group in solution or bound to the enzyme. The K(i) value for the inhibitor decreases outside the optimum pH range for catalysis, indicating that binding of D-glucal is tighter with incorrectly ionized forms of the complex between the enzyme and alpha-D-glucose 1-phosphate. Each molecule of trehalose phosphorylase contains one Mg(2+) that is non dissociable in the presence of metal chelators. Measurements of the (26)Mg(2+)/(24)Mg(2+) ratio in the solvent and on the enzyme by using inductively coupled plasma MS show that exchange of metal ion between protein and solution does not occur at measurable rates. Tryptic peptide mass mapping reveals close structural similarity between trehalose phosphorylases from basidiomycete fungi. PMID- 11389684 TI - Amine oxidase substrates mimic several of the insulin effects on adipocyte differentiation in 3T3 F442A cells. AB - We have previously reported that substrates of monoamine oxidase (MAO) and semicarbazide-sensitive amine oxidase (SSAO) exert short-term insulin-like effects in rat adipocytes, such as stimulation of glucose transport. In the present work, we studied whether these substrates could also mimic long-term actions of insulin. Adipose differentiation of 3T3 F442A cells, which is highly insulin-dependent, served as a model to test the effects of sustained administration of amine oxidase substrates. Daily treatment of confluent cells with 0.75 mM tyramine (a substrate of MAO and SSAO) or benzylamine (a substrate of SSAO) over 1 week caused the acquisition of typical adipocyte morphology. The stimulation of protein synthesis and triacylglycerol accumulation caused by tyramine or benzylamine reached one half of that promoted by insulin. This effect was insensitive to pargyline (an MAO inhibitor), but was inhibited by semicarbazide (an SSAO inhibitor) and by N-acetylcysteine (an antioxidant agent), suggesting the involvement of the H(2)O(2) generated during SSAO-dependent amine oxidation. Chronic administration of amine oxidase substrates also induced the emergence of adipose conversion markers, such as aP2, glycerol-3-phosphate dehydrogenase, the glucose transporter GLUT4, and SSAO itself. Moreover, cells treated with amines acquired the same insulin sensitivity regarding glucose transport as adipocytes classically differentiated with insulin. In all, most of the adipogenic effects of amines were additive to insulin. Our data reveal that amine oxidase substrates partially mimic the adipogenic effect of insulin in cultured preadipocytes. Furthermore, they suggest that SSAO not only represents a novel late marker of adipogenesis, but could also be directly involved in the triggering of terminal adipocyte differentiation. PMID- 11389685 TI - A mitochondrial uncoupling artifact can be caused by expression of uncoupling protein 1 in yeast. AB - Uncoupling protein 1 (UCP1) from mouse was expressed in yeast and the specific (GDP-inhibitable) and artifactual (GDP-insensitive) effects on mitochondrial uncoupling were assessed. UCP1 provides a GDP-inhibitable model system to help interpret the uncoupling effects of high expression in yeast of other members of the mitochondrial carrier protein family, such as the UCP1 homologues UCP2 and UCP3. Yeast expressing UCP1 at modest levels (approx. 1 microg/mg of mitochondrial protein) showed no growth defect, normal rates of chemically uncoupled respiration and an increased non-phosphorylating proton conductance that was completely GDP-sensitive. The catalytic-centre activity of UCP1 in these yeast mitochondria was similar to that in mammalian brown-adipose-tissue mitochondria. However, yeast expressing UCP1 at higher levels (approx. 11 microg/mg of mitochondrial protein) showed a growth defect. Their mitochondria had depressed chemically uncoupled respiration rates and an increased proton conductance that was partly GDP-insensitive. Thus, although UCP1 shows native behaviour at modest levels of expression in yeast, higher levels (or rates) of expression can lead to an uncoupling that is not a physiological property of the native protein and is therefore artifactual. This observation might be important in the interpretation of results from experiments in which the functions of UCP1 homologues are verified by their ability to uncouple yeast mitochondria. PMID- 11389686 TI - Identification of novel beta-mannan- and beta-glucan-binding modules: evidence for a superfamily of carbohydrate-binding modules. AB - Many glycoside hydrolases, which degrade long-chain carbohydrate polymers, possess distinct catalytic modules and non-catalytic carbohydrate-binding modules (CBMs). On the basis of conserved protein secondary structure, we describe here the identification and experimental characterization of novel type of mannanase associated mannan-binding module and also characterization of two CBM family 4 laminarinase-associated beta-glucan-binding modules. These modules are predicted to belong to a superfamily of CBMs which include families 4, 16, 17, 22 and a proposed new family, family 27. PMID- 11389687 TI - Role of ergothioneine on S-nitrosoglutathione catabolism. AB - Ergothioneine (ESH) is a low-molecular-mass thiol present in millimolar concentrations in a limited number of tissues, including erythrocytes, kidney, seminal fluid and liver; however, its biological function is still unclear. In the present study we investigated the role of ESH in the catabolism of S nitrosoglutathione (GSNO). The results show that: (1) GSNO decomposition is strongly influenced by ESH (k"=0.178+/-0.032 M(-1) x s(-1)); (2) ammonia is the main nitrogen-containing compound generated by the reaction; and (3) nitrite is practically absent under both aerobic and anaerobic conditions. These findings are markedly different from those reported for the GSH-induced decomposition of GSNO, in which the nitrogen-containing end products are nitrite, ammonia and nitrous oxide (N(2)O) under aerobic conditions but nitrite, ammonia, nitric oxide (NO) and small quantities of hydroxylamine under anaerobic conditions. Considering the high concentration of ESH in specific cells, the reaction with GSNO should be considered as an important molecular event occurring in the cell. PMID- 11389688 TI - Response of neurons to an irreversible inhibition of endoplasmic reticulum Ca(2+) ATPase: relationship between global protein synthesis and expression and translation of individual genes. AB - In the physiological state, there appears to be a regulatory link between endoplasmic reticulum (ER) Ca(2+) homoeostasis and the initiation of neuronal protein synthesis. Exposing neuronal cell cultures to thapsigargin (Tg), an irreversible inhibitor of sarcoplasmic/ER Ca(2+)-ATPase (SERCA), induced an almost complete suppression of protein synthesis, which recovered to approx. 60% of control 24 h after Tg exposure. This is an experimental model where the regulatory link between the initiation of protein synthesis and ER Ca(2+) homoeostasis recovers, despite an irreversible suppression of SERCA activity [Doutheil, Treiman, Oschlies and Paschen (1999) Cell Calcium 25, 419--428]. The model was used to investigate the relationship between transcription and translation of various stress genes that respond to conditions causing graded suppression of protein synthesis. Expression patterns revealed three groups of genes. The mRNA levels of genes responding to conditions of ER stress (grp78, grp94, gadd34 and gadd153) were increased up to 200-fold after Tg exposure, whereas those coding for ER-resident proteins (SERCA 2b and Bcl-2) were increased up to 6-fold in treated cultures, and those coding for cytoplasmic proteins (heat shock protein 70 and p67) were increased only 2--4-fold. Analysis of translation of these mRNAs suggests an imbalance in the synthesis of apoptosis-inducing (GADD153) and tolerance-activating (GRP78 and Bcl-2) proteins after blocking of the ER Ca(2+) pump. The observation that the relationship between Tg-induced changes in mRNA and protein levels varied considerably for the various genes studied implies that translation of the respective transcripts is differently regulated under conditions causing graded suppression of global protein synthesis. Detailed analysis of the response of neuronal cells to transient disturbance of ER Ca(2+) homoeostasis may help to elucidate the mechanisms underlying neuronal cell injury in those neurological disorders in which an impairment of ER function is thought to contribute to the pathological process of deterioration. PMID- 11389689 TI - Highly sensitive and selective fluorescence assays for rapid screening of endothelin-converting enzyme inhibitors. AB - The highly potent vasoconstrictor peptide endothelin (ET) is generated from an inactive precursor, big endothelin (bET), by endothelin-converting enzyme (ECE). ECE is a phosphoramidon-sensitive zinc metallopeptidase, which is closely related to neprilysin (neutral endopeptidase). It is possible that compounds which inhibit the formation of ET may be used as new drugs for the treatment of cardiovascular diseases. Such an approach requires a fast, simple and selective assay to measure ECE activity, allowing rapid screening of inhibitors. We describe here two new ECE substrates based on the concept of 'intramolecularly quenched fluorescence' which may fulfill this aim. One, S(1) [Pya(21)-Nop(22)-bET 1(19--35)], is the (19--35) fragment of the natural peptide big-ET-1(1--38), which is modified by introducing the fluorescent amino acid, pyrenylalanine (Pya), in position 21 and a quencher, p-nitrophenylalanine (Nop), in position 22. The second substrate (S(2)) is a small peptide, Ac-Ser-Gly-Pya-Lys-Ala-Phe-Ala Nop-Gly-Lys-NH(2), from a biased substrate peptide library. The recombinant, hECE 1c, cleaved both Pya(21)-Nop(22)-bET-1(19--35) and the natural substrate selectively between residues 21 and 22, whereas cleavage occurred between alanine and phenylalanine in the small peptide. In both cases, this generated intense fluorescence emission. The synthesis and kinetic parameters of these substrates are described. These assays, which can be used directly on tissue homogenates, are the most sensitive and selective described to date for ECE, and are easily automated for a high-throughput screening of inhibitors. PMID- 11389690 TI - Superior role of apolipoprotein B48 over apolipoprotein B100 in chylomicron assembly and fat absorption: an investigation of apobec-1 knock-out and wild-type mice. AB - Editing of apolipoprotein (apo)-B100 mRNA to yield apo-B48 is a specific and developmentally regulated step in enterocytes of mammals. However, the functional significance of this step is not known. Since mice containing only apo-B100 have not been documented to exhibit any difference in intestinal fat absorption from wild-type mice, the evolutionary advantage of apoB mRNA editing has been questioned. In the present study, we have compared fat absorption and chylomicron assembly in apobec-1 knock-out (KO) or wild-type (WT) mice subjected to different dietary manipulations: low-fat chow, a fat-enriched 'Western' diet and overnight fasting. Experiments in vivo and in vitro revealed differences in the ability of KO and WT enterocytes to assemble and secrete chylomicrons under different dietary conditions. After overnight fasting, chylomicron secretion is reduced considerably in KO compared with WT enterocytes. This is not due to reduced synthesis of apo-B or triacylglycerol (TAG), but appears to be a result of impaired assembly of chylomicrons, so that triacylglycerol accumulates in the enterocytes. After feeding with fat, secretion of chylomicrons enriched in pre existing TAG is stimulated in KO compared with WT mice. In the present study, we have documented for the first time that apo-B100 is considerably less efficient than apo-B48 in exerting its role in the early stage of chylomicron assembly, which is rate-limiting under conditions of low dietary fat. However, this impairment is overcome by increased TAG stores that stimulate later stages in assembly, which are rate-limiting in the fat-fed state. apo-B mRNA editing may result in more efficient fat absorption, specifically under conditions of food shortage or low-fat content, and thus provide an evolutionary advantage. PMID- 11389691 TI - Interaction of human thymidine kinase 1 with p21(Waf1). AB - The overexpression of the cyclin-dependent kinase (CDK) inhibitor p21(Waf1) can inhibit cell proliferation, which is mediated by direct binding to CDK and proliferating-cell nuclear antigen. In this study, we demonstrated that human cytosolic thymidine kinase 1 (TK1) polypeptide can form a complex with p21(Waf1). The C-terminal domain of p21(Waf1) appeared to interact with the TK1 polypeptide, but, despite the inhibitory function of p21(Waf1), their association did not alter TK1 functional activity. However, overexpression of TK1 overcame p21(Waf1) mediated growth suppression and blocked the association of CDK2 with p21(Waf1), suggesting that TK1 interferes with the inhibitory function of p21(Waf1). Based on these results, we here propose that the molecular function of p21(Waf1) in cells can be perturbed through its interaction with another cellular protein, TK1. PMID- 11389692 TI - Proteasomes are a target of the anti-tumour drug vinblastine. AB - Proteasomes, the proteolytic machinery of the ubiquitin/ATP-dependent pathway, have a relevant role in many processes crucial for cell physiology and cell cycle progression. Proteasome inhibitors are used to block cell cycle progression and to induce apoptosis in certain cell lines. Here we examine whether proteasomal function is affected by the anti-tumour drug vinblastine, whose cytostatic action relies mainly on the disruption of mitotic spindle dynamics. The effects of vinblastine on the peptidase activities of human 20 S and 26 S proteasomes and on the proteolytic activity of 26 S proteasome were assessed in the presence of specific fluorogenic peptides and (125)I-lysozyme-ubiquitin conjugates respectively. The assays of ubiquitin-protein conjugates and of inhibitory kappa B alpha (I kappa B alpha), which are characteristic intracellular proteasome substrates, by Western blotting on lysates from HL60 cells incubated with or without vinblastine, illustrated the effects of vinblastine on proteasomes in vivo. We also evaluated the effects of vinblastine on the signal-induced degradation of I kappa B alpha. Vinblastine at 3--110 microM reversibly inhibited the chymotrypsin-like activity of the 20 S proteasome and the trypsin-like and peptidyl-glutamyl-peptide hydrolysing activities of both proteasomes, but only at 110 microM vinblastine was the chymotrypsin-like activity of the 26 S proteasome inhibited; furthermore, at 25--200 microM the drug inhibited the degradation of ubiquitinated lysozyme. In HL60 cells exposed for 6 h to 0.5--10 microM vinblastine, the drug-dose-related accumulation of polyubiquitinated proteins, as well as that of a high-molecular-mass form of I kappa B alpha, occurred. Moreover, vinblastine impaired the signal-induced degradation of I kappa B alpha. Cell viability throughout the test was approx. 95%. Proteasomes can be considered to be a new and additional vinblastine target. PMID- 11389693 TI - Cholesterol-dependent interaction of syncollin with the membrane of the pancreatic zymogen granule. AB - Syncollin is a protein of the pancreatic zymogen granule that was isolated through its ability to bind to syntaxin. Despite this in vitro interaction, it is now clear that syncollin is present on the luminal side of the zymogen granule membrane. Here we show that there are two pools of syncollin within the zymogen granule: one free in the lumen and the other tightly associated with the granule membrane. When unheated or cross-linked samples of membrane-derived syncollin are analysed by SDS/PAGE, higher-order forms are seen in addition to the monomer, which has an apparent molecular mass of 16 kDa. Extraction of cholesterol from the granule membrane by treatment with methyl-beta-cyclodextrin causes the detachment of syncollin, and this effect is enhanced at a high salt concentration. Purified syncollin is able to bind to brain liposomes at pH 5.0, but not at pH 11.0, a condition that also causes its extraction from granule membranes. Syncollin binds only poorly to dioleoyl phosphatidylcholine liposomes, but binding is dramatically enhanced by the inclusion of cholesterol. Finally, cholesterol can be co-immunoprecipitated with syncollin. We conclude that syncollin is able to interact directly with membrane lipids, and to insert into the granule membrane in a cholesterol-dependent manner. Membrane-associated syncollin apparently exists as a homo-oligomer, possibly consisting of six subunits, and its association with the membrane may be stabilized by electrostatic interactions with either other proteins or phospholipids. PMID- 11389694 TI - Overproduction, purification and novel redox properties of the dihaem cytochrome c, NapB, from Haemophilus influenzae. AB - The napB gene of the pathogenic bacterium Haemophilus influenzae encodes a dihaem cytochrome c, the small subunit of a heterodimeric periplasmic nitrate reductase similar to those found in other bacteria. In order to obtain sufficient protein for biophysical studies, we aimed to overproduce the recombinant dihaem protein in Escherichia coli. Initial expression experiments indicated that the NapB signal peptide was not cleaved by the leader peptidase of the host organism. Apocytochrome was formed under aerobic, semi-aerobic and anaerobic growth conditions in either Luria--Bertani or minimal salts medium. The highest amounts of apo-NapB were produced in the latter medium, and the bulk was inserted into the cytoplasmic membrane. The two haem groups were covalently attached to the pre apocytochrome only under anaerobic growth conditions, and with 2.5 mM nitrite or at least 10 mM nitrate supplemented to the minimal salts growth medium. In order to obtain holocytochrome, the gene sequence encoding mature NapB was cloned in frame with the E. coli ompA (outer membrane protein A) signal sequence. Under anaerobic conditions, NapB was secreted into the periplasmic space, with the OmpA signal peptide being correctly processed and with both haem c groups attached covalently. Unless expressed in the DegP-protease-deficient strain HM125, some of the recombinant NapB polypeptides were N-terminally truncated as a result of proteolytic activity. Under aerobic growth conditions, co-expression with the E. coli ccm (cytochrome c maturation) genes resulted in a higher yield of holocytochrome c. The pure recombinant NapB protein showed absorption maxima at 419, 522 and 550 nm in the reduced form. The midpoint reduction potentials of the two haem groups were determined to be -25 mV and -175 mV. These results support our hypothesis that the Nap system fulfils a nitrate-scavenging role in H. influenzae. PMID- 11389695 TI - Transcriptional activities of p73 splicing variants are regulated by inter variant association. AB - p73 has been identified as a gene that encodes a protein with significant identity with the tumour suppressor p53. The main structural difference between p73 and p53 is the additional C-terminal region of p73. Six isoforms of p73 with differing C-terminal structures, alpha, beta, gamma, delta, epsilon and xi, have been reported. These variants differ in transcriptional activity on p53 responsive promoters. Here we report a possible mechanism of transcriptional activation by p73 splicing variants. C-terminal deletion mutants of p73 alpha showed a significantly higher level of transcriptional activity than wild-type p73 alpha, suggesting that the C-terminal structure of p73 alpha functions to repress the transcriptional activity of p73 alpha. The results of immunoprecipitation assays and two-hybrid assays in mammalian cells showed that the p73 variants interacted with each other, but not with p53. The transcriptional activity of p73 beta was reduced by co-expression with either p73 alpha or p73 epsilon, which bears an identical C-terminal structure to p73 alpha. Co-expression of the C-terminal portion of p73 alpha or p73 epsilon with p73 beta also resulted in reduced transcriptional activity. Moreover, we observed that the level of endogenous p21 protein induced by p73 beta was decreased by co expression of full-length p73 epsilon or the C-terminal region of p73 alpha or p73 epsilon. These observations suggest that p73-mediated gene expression is regulated by the interactions of p73 splicing variants in the cell. PMID- 11389696 TI - Conjugation of an antibody Fv fragment to a virus coat protein: cell-specific targeting of recombinant polyoma-virus-like particles. AB - The development of cell-type-specific delivery systems is highly desirable for gene-therapeutic applications. Current virus-based vector systems show broad cell specificity, which results in the need to restrict the natural tropism of these viral systems. Here we demonstrate that tumour-cell-specific virus-like particles can be functionally assembled in vitro from recombinant viral coat protein expressed in Escherichia coli. The insertion of a negatively charged peptide in the HI loop of polyoma VP1 interferes with the binding of VP1 to the natural recognition site on mammalian cells and also serves as an adapter for the coupling of antibody fragments that contain complementary charged fusion peptides. A recombinant antibody fragment of the tumour-specific anti-(Lewis Y) antibody B3 could be coupled to the mutant VP1 by engineered polyionic peptides and an additional disulphide bond. With this system an entirely recombinant cell specific delivery system assembled in vitro could be generated that transfers genes preferentially to cells presenting the tumour-specific antigen on the cell surface. PMID- 11389697 TI - Thrombopoietin stimulates cortactin translocation to the cytoskeleton independently of tyrosine phosphorylation. AB - Cortactin is an F-actin-binding protein expressed in platelets. During aggregation by thrombin, cortactin associates with Src, is tyrosine phosphorylated, and then translocates to the cytoskeleton. It is also found to associate with Syk during platelet shape change. Since cortactin undergoes tyrosine phosphorylation in platelets activated by thrombopoietin (TPO) that exhibit neither shape change nor aggregation, we investigated whether it could also relocalize to the detergent-insoluble fraction. We demonstrate that cortactin was present as a tyrosine-phosphorylated protein and co-localized with Syk in the Triton X-100-insoluble fraction of TPO-activated platelets. TPO stimulated Syk activation and association with cortactin. Conversely, cortactin associated with the kinases, Syk and Src. Cortactin tyrosine phosphorylation was blocked by Syk kinase inhibitor, piceatannol or Src family kinase inhibitor, PP2, suggesting that it depends on these two kinases. However, piceatannol or PP2 did not prevent cortactin translocation to the detergent-insoluble fraction. These data suggest that tyrosine phosphorylation is not required for cortactin translocation to the detergent-insoluble compartment. Furthermore, TPO activates, through its receptor c-Mpl, a signalling pathway to the cytoskeleton. PMID- 11389698 TI - Effect of iron deficiency on placental transfer of iron and expression of iron transport proteins in vivo and in vitro. AB - Maternal iron deficiency during pregnancy induces anaemia in the developing fetus; however, the severity tends to be less than in the mother. The mechanism underlying this resistance has not been determined. We have measured placental expression of proteins involved in iron transfer in pregnant rats given diets with decreasing levels of iron and examined the effect of iron deficiency on iron transfer across BeWo cell layers, a model for placental iron transfer. Transferrin receptor expression was increased at both mRNA and protein levels. Similarly, expression of the iron-responsive element (IRE)-regulated form of the divalent metal transporter 1 (DMT1) was also increased. In contrast, the non-IRE regulated isoform showed no change in mRNA levels. Protein levels of DMT1 increased significantly. Iron efflux is thought to be mediated by the metal transporter protein, IREG1/ferroportin1/MTP1, and oxidation of Fe(II) to Fe(III) prior to incorporation into fetal transferrin is carried out by the placental copper oxidase. Expression of IREG1 was not altered by iron deficiency, whereas copper oxidase activity was increased. In BeWo cells made iron deficient by treatment with desferrioxamine ('deferioxamine'), iron accumulation from iron transferrin increased, in parallel with increased expression of the transferrin receptor. At the same time, iron efflux also increased, showing a higher flux of iron from the apical to the basolateral side. The data show that expression of placental proteins of iron transport are up-regulated in maternal iron deficiency, resulting in an increased efficiency of iron flux and a consequent minimization of the severity of fetal anaemia. PMID- 11389699 TI - Identification of a new subfamily of sulphotransferases: cloning and characterization of canine SULT1D1. AB - Sulphation is an important conjugation pathway in drug metabolism that has been studied in several species including humans. However, few studies have been performed using the dog as a subject. In this report we describe the cloning and characterization of a canine cytosolic sulphotransferase (SULT). The overall primary structure of this enzyme is very similar to that of a rat phenol sulphating enzyme found in the EMBL Database and to a mouse SULT termed amine-N sulphotransferase (81% identity). The expressed canine SULT conjugates small phenols and aromatic amines such as dopamine, minoxidil, p-nitrophenol and 5 hydroxytryptamine, but not dehydroepiandrosterone or beta-oestradiol. These results are in agreement with the results reported for the mouse SULT. In contrast with the mouse enzyme, the canine SULT does not conjugate eicosanoid compounds, i.e. prostaglandins, thromboxane B(2) or leukotriene E(4). The canine SULT is expressed at high levels in the colon of both genders; it is also expressed in the small intestine, kidney and liver. Furthermore, because the canine, mouse and rat SULT forms exhibit significant sequence identity (more than 80%), they seem to represent a distinct group in the SULT family tree. This suggestion is strengthened by the low identity with other SULTs. The subfamily that is most similar to this new group is SULT1A, with approx. 60% similarity. However, the mouse and canine enzymes are not characterized by the efficient sulphation of p-nitrophenol, dopamine, beta-oestradiol or oestrone. Thus these results seem to exclude them from the SULT1A subfamily. We therefore propose a new subfamily in the phenol SULT family, designated SULT1D, and consequently the canine enzyme is termed SULT1D1. PMID- 11389700 TI - Inhibition of peroxisome-proliferator-activated receptor (PPAR)alpha by MK886. AB - Although MK886 was originally identified as an inhibitor of 5-lipoxygenase activating protein (FLAP), recent data demonstrate that this activity does not underlie its ability to induce apoptosis [Datta, Biswal and Kehrer (1999) Biochem. J. 340, 371--375]. Since FLAP is a fatty-acid binding protein, it is conceivable that MK886 may affect other such proteins. A family of nuclear receptors that are activated by fatty acids and their metabolites, the peroxisome proliferator-activated receptors (PPARs), have been implicated in apoptosis and may represent a target for MK886. The ability of MK886 to inhibit PPAR-alpha, beta and -gamma activity was assessed using reporter assay systems (peroxisome proliferator response element--luciferase). Using a transient transfection system in monkey kidney fibroblast CV-1 cells, mouse keratinocyte 308 cells and human lung adenocarcinoma A549 cells, 10--20 microM MK886 inhibited Wy14,643 activation of PPAR alpha by approximately 80%. Similar inhibition of PPAR alpha by MK886 was observed with a stable transfection reporter system in CV-1 cells. Only minimal inhibitory effects were seen on PPAR beta and PPAR gamma. MK886 inhibited PPAR alpha by a non-competitive mechanism as shown by its effects on the binding of arachidonic acid to PPAR alpha protein, and a dose-response study using a transient transfection reporter assay in COS-1 cells. An assay assessing PPAR ligand-receptor interactions showed that MK886 prevents the conformational change necessary for active-complex formation. The expression of keratin-1, a protein encoded by a PPAR alpha-responsive gene, was reduced by MK886 in a culture of mouse primary keratinocytes, suggesting that PPAR inhibition has functional consequences in normal cells. Although Jurkat cells express all PPAR isoforms, various PPAR alpha and PPAR gamma agonists were unable to prevent MK886-induced apoptosis. This is consistent with MK886 functioning as a non-competitive inhibitor of PPAR alpha, but may also indicate that PPAR alpha is not directly involved in MK886-induced apoptosis. Although numerous PPAR activators have been identified, the results show that MK886 can inhibit PPAR alpha, making it the first compound identified to have such an effect. PMID- 11389701 TI - Cloning and characterization of human liver cytosolic beta-glycosidase. AB - Cytosolic beta-glucosidase (EC 3.2.1.21) from mammalian liver is a member of the family 1 glycoside hydrolases and is known for its ability to hydrolyse a range of beta-D-glycosides, including beta-D-glucoside and beta-D-galactoside. We therefore refer to this enzyme as cytosolic beta-glycosidase. We cloned the cDNA encoding the human cytosolic beta-glycosidase by performing PCR on cDNA prepared from total human liver RNA. Specific primers were based on human expressed sequence tags found in the expressed sequence tag database. The cloned cDNA contained 1407 nt with an open reading frame encoding 469 amino acid residues. Amino acid sequence analysis indicates that human cytosolic beta-glycosidase is most closely related to lactase phlorizin hydrolase and klotho protein. The enzyme was characterized by using cell lysates of COS-7 cells transfected with a eukaryotic expression vector containing the cDNA. The biochemical, kinetic and inhibition properties of the cloned enzyme were found to be identical with those reported for the enzyme purified from human liver. PMID- 11389703 TI - Treatment of severe acute graft-versus-host disease with anti-thymocyte globulin. AB - Severe acute graft-versus-host disease (GVHD) is one of the major complications after haematopoietic stem-cell transplantation (HSCT). Treatment of severe GVHD is difficult and the condition is often fatal. One proposed method of improving the therapy is to include anti-thymocyte globulin (ATG). Here, we will report our results in 29 patients using ATG as part of treatment for severe steroid resistant acute GVHD. Four patients suffered from grade II, 13 from grade III and 12 from grade IV GVHD. Median time to grade II GVHD was 24 d (range 7-91 d) and to grade III was 29 d (range 8-55 d) after HSCT. Five different ATG preparations were used, rabbit ATG (R-ATG), BMA 031, OKT3, ATG-Fresenius and Thymoglobuline. All patients had skin involvement, 26 also had gut involvement and 25 had liver involvement. The rate of response to treatment was best in skin involvement (72%), while liver and gut involvement showed lower response rates (38%). Eleven patients survived more than 90 d, 7 of them developed chronic GVHD, 1 developed mild GVHD, 1 developed moderate GVHD and 5 developed severe GVHD. Survival at 100 d was 37% and at 1 yr it was 12%. Most patients died of GVHD, with virus or fungal infections as contributing causes of death. To conclude, treatment of severe acute GVHD is difficult and ATG, in our hands, adds nothing to conventional pharmacological treatment. PMID- 11389704 TI - Peri-operative cardiac morbidity in kidney transplant recipients: incidence and risk factors. AB - BACKGROUND: Renal transplant recipients are known to be at increased risk for developing cardiac disease. In both general and peripheral vascular surgery, pre operative risk stratification (and intervention when indicated) has decreased the incidence of peri-operative cardiac complications. In this study, we set out to identify subsets of patients at high risk for peri-operative cardiac complications after a renal transplant. METHODS: We retrospectively reviewed the records of 2694 adult renal transplants performed at the University of Minnesota between January 1, 1985 and December 31, 1998. We determined the incidence of peri-operative (within 30 d post-transplant) cardiac complications, including myocardial infarction (MI). Risk factors for the development of these complications were determined by multivariate analysis. RESULTS: We found 163 peri-operative cardiac complications, for an overall incidence of 6.1%. Specific cardiac complications included MI (n=43, 1.6%), arrhythmia (n=74, 2.7%), angina (n=31, 1.2%), cardiac arrest (n=13, 0.5%), and congestive heart failure (n= 2, 0.1%). By multivariate analysis, significant risk factors for any cardiac complication were age> or =50 yr (relative risk (RR)=3.0, p=0.0001) and pre transplant cardiac disease (RR=3.3, p=0.0001). Not significant were diabetes mellitus (DM), cadaver donor source, pre-transplant dialysis, a history of smoking, and hypertension. Significant risk factors for peri-operative MI were age> or =50 yr, pre-existing cardiac disease, and DM. Diabetic patients with pre existing cardiac disease were at especially high risk for peri-operative cardiac events. CONCLUSIONS: Patients>50 yr and those with pre-existing cardiac disease, especially if diabetic, are at significantly increased risk for developing peri operative cardiac complications after a renal transplant. Such patients require aggressive pre-operative investigations, which may include coronary angiography, to decrease the risk of post-transplant complications. PMID- 11389705 TI - Low rejection rates with tacrolimus-based dual and triple regimens following liver transplantation. AB - We studied the outcome of 345 liver transplant patients who received tacrolimus based immunosuppressive therapy either as a dual regimen (with corticosteroids, n=172) or as a triple regimen (with corticosteroids and azathioprine, n=173) for 3 months after transplantation (3-month cohort). A further analysis was conducted for the first 195 patients randomised (dual n=100, triple n=95) who were followed up for 12 months after transplantation (12-month cohort). For the 3-month cohort, patient survival was 90.7% (dual) and 91.9% (triple), graft survival after 3 months was 88.4% (dual therapy) and 89.6% (triple therapy). Acute rejections were experienced by 67/172, 39.0% of patients on dual therapy and by 60/173, 34.7% of patients on triple therapy; corticosteroid-resistant rejections were reported in 9 patients (5.2%) in either treatment group. The overall safety profile was similar for the two treatment groups. Significant differences, however, were found for thrombocytopenia (dual 13/172, 7.6%, triple 37/173, 21.4%, p<0.001) and leukopenia (dual 4/172, 2.3%, triple 24/173, 13.9%, p<0.001). For the 12-month cohort, patient survival was 85.6% (dual) and 88.4% (triple) after 1 year. Graft survival was 81.7% (dual) and 85.2% (triple) 12 months after transplantation. Acute rejections were reported for 38/100, 38.0% of patients on dual therapy and 36/95, 37.9% of patients on triple therapy, corticosteroid-resistant rejections were 7/100, 7.0% (dual) and 7/95, 7.4% (triple) of patients. In the 12-month cohort, no significant differences in the safety profiles of the treatment groups were found. We conclude that both tacrolimus-based dual and triple drug regimens provide effective and safe immunosuppression following orthotopic liver transplantation. PMID- 11389706 TI - Allogeneic bone marrow transplantation for chronic myeloid leukemia: a retrospective study of busulfan-cytoxan versus total body irradiation-cytoxan as preparative regimen in Koreans. AB - From January 1990 to December 1997, 53 Korean patients with chronic myeloid leukemia (CML) receiving bone marrow transplantation (BMT) from human leucocyte antigen (HLA)-identical sibling donors conditioned with either busulfan and cyclophosphamide (BU/CY regimen) or total body irradiation and cyclophosphamide (TBI/CY regimen) were compared retrospectively. Transplantation-related mortality was 19% in BU/CY and 12% in TBI/CY, and early death (<100 d) occurred in 3 patients conditioned with BU/CY. Grade II-IV acute graft-versus-host disease (GVHD) was 9% of BU/CY and 52% of TBI/CY patients. Overall incidence of chronic GVHD was 50% of BU/CY and 52% of TBI/CY patients. In patients with chronic phase, 5-yr overall survival was 73% in the BU/CY group compared with 87% in the TBI/CY group (p=NS), and overall disease-free survival was 75% in the BU/CY group and 59% in the TBI/CY group (p=NS). So far, with a median follow-up of 45 months, 11 patients have relapsed; three relapses occurred after BU/CY and 8 after TBI/CY. The actuarial 5-yr relapse rate was 15% after BU/CY, 34% after TBI/CY (p=0.46). For patients transplanted in chronic phase within 1 yr after diagnosis, there was a clear trend for a lower relapse rate in the BU/CY group (5-yr relapse rate 0%) compared with the TBI/CY group (5-yr relapse rate 30%). The BU/CY group had similar BMT-related toxicity and similar overall survival and showed a clear trend of low relapse compared with the TBI/CY group. Therefore, BU/CY is an acceptable alternative for patients with CML during HLA-identical sibling allogeneic BMT. PMID- 11389707 TI - Effect of lovastatin, an HMG CoA reductase inhibitor, on acute renal allograft rejection. AB - 3-Hydroxy-3-methyl glutaryl coenzyme A (HMG CoA) reductase inhibitors are established anti-lipidemic agents. They also exert immunomodulatory effects. Two recent reports suggest that pravastatin may be useful in decreasing the incidence and severity of acute rejections (ARs) in heart and kidney transplant recipients. We undertook this prospective, randomized, placebo-controlled, double blind trial to investigate the effect of lovastatin on acute renal allograft rejection. Sixty five consecutive, one-haplotype-matched, living related first renal transplant recipients were randomized to receive either lovastatin 20 mg/d or placebo for 3 months, in addition to cyclosporine, azathioprine, and steroids. Lipid levels, AR episodes, and liver and muscle enzymes were followed for 3 months post transplant. At the end of the study period, lovastatin had successfully controlled lipid levels. However, there was no effect on AR episodes (15.15% in the treatment group vs. 18.75% in the placebo group). PMID- 11389708 TI - Pharmacokinetics of intravenous mycophenolate mofetil after allogeneic blood stem cell transplantation. AB - BACKGROUND: Mycophenolate mofetil (MMF) has shown synergistic effects in combination with cyclosporin A (CsA) in prevention of acute graft versus host disease (GvHD) after allogeneic blood stem cell transplantation (BSCT) in preclinical animal models. After having measured low plasma levels of the active metabolite mycophenolic acid (MPA) in recipients of allogeneic blood stem cell transplants after oral administration of MMF, we initiated a phase I/II study evaluating different dose levels of the intravenous (i.v.) formulation together with standard dose CsA. METHODS: A total of 15 patients received i.v. MMF in two split doses for 21 d after allogeneic BSCT from related (n=9) and unrelated (n=6) donors. Total daily doses of 25, 28, 31 and 34 mg/kg were investigated in 3-5 patients at each dose level. Plasma concentrations of MPA and its metabolite mycophenolic acid glucuronide (MPAG) were measured by high-performance liquid chromatography (HPLC). RESULTS: Mean trough blood levels of MPA ranged between 68.8 and 340 ng/mL with a median of 146.7 ng/mL. The mean MPA AUC0-12 h after first dose ranged between 19349+/-5087 ng * h/mL and 25705+/-3042 ng * h/mL and correlated with the dose level of MMF. The incidence of acute GvHD>grade I was 40%. No dose limiting toxicities were observed. CONCLUSIONS: The application of i.v. MMF is safe at a weight-adjusted dose between 25 and 34 mg/kg after allogeneic BSCT. The measured trough blood levels of MPA in patients after BSCT were ten times lower than in healthy volunteers. The toxicity induced by the conditioning therapy seems to negatively influence the pharmacokinetic behavior of MMF, MPA and MPAG. PMID- 11389709 TI - Transforming growth factor-beta1 expression in early biopsy specimen predicts long-term graft function following pediatric renal transplantation. AB - The main cause of late graft loss or declining long-term graft function is chronic allograft nephropathy (CAN), characterized by progressive interstitial fibrosis. Transforming growth factor (TGF)-beta1 plays a key role in fibrogenesis. We immunohistochemically investigated whether the degree of TGF beta1 expression in early biopsy specimens routinely obtained from stable allografts at 100 d could predict fibrosis and graft dysfunction in the late phase. Patients were children with grafts from related donors. We immunohistochemically determined intracellular and extracellular expression of TGF-beta1 in the graft using LC antibody (LC) for intracellular TGF-beta1 and CC antibody (CC) for extracellular TGF-beta1. The change in creatinine clearance between 100 d and 3 yr after transplantation (DeltaCcr) was used as an index of long-term graft function. We also used image analysis to calculate the relative area involved by interstitial fibrosis in the trichrome-stained section of graft biopsy specimens at 100 d and 3 yr, designating the change as DeltaFI. DeltaCcr was -4.2+/-9.4 mL/min in subjects with minimal early immunoreactivity for CC and 20.5+/-15.9 mL/min in subjects with strong reactivity (p<0.05). DeltaCcr was 14.5+/-18.6 mL/min in subjects with minimal early immunoreactivity for LC and 11.7+/-12.8 mL/min in those with strong reactivity. DeltaFI in subjects with minimal CC reactivity (1.28+/-4.11%) tended to be lower than that in subjects with strong reactivity (8.45+/-15.47%). Neither fibrosis at 100 d nor DeltaFI differed between subjects with minimal and strong LC reactivity. Thus, strong extracellular TGF-beta1 expression in grafts at 100 d after transplantation is associated with a long-term decline in graft function and tends to be associated with increased graft fibrosis at 3 yr. PMID- 11389710 TI - Prospective study of microchimerism in renal allograft recipients: association between HLA-DR matching, microchimerism and acute rejection. AB - The presence of donor-derived hematopoietic cells in blood and various tissues of the organ recipients, termed allogeneic microchimerism, has been considered to play an essential role in establishment of organ acceptance. In this study, we prospectively determined the presence of peripheral blood microchimerism (PBM) in 20 male-to-female renal allograft recipients up to 30 months post transplantation. Recipients were categorized according to the pattern of microchimerism into microchimeric and nonmicrochimeric groups, and then state of human leukocyte antigens (HLA) Class II (DR/DQ) matching, episodes of acute rejection, age at transplantation, renal function, and history of blood transfusion were compared. DNA was extracted from donor, pre-transplant, and post transplant (1 wk; 1, 3, 6, 12, 18, 24, and 30 months) peripheral blood samples. We analyzed PBM using nested polymerase chain reaction (PCR) amplification specific for the SRY region of the Y chromosome with a sensitivity up to 1:1 000 000. Microchimerism was detected in 13 (65%) of 20 recipients at various intervals. The highest frequency of microchimerism was at 1 wk (55%). Among microchimeric recipients, none were positive on all post-transplant analyses. Interestingly, nonmicrochimeric cases were negative throughout the study. The three recipients with an episode of acute rejection during the first week after transplantation were all in the nonmicrochimeric group with completely mismatched HLA-DR antigens. HLA-DR incompatibility was significantly lower (t-test, p<0.05) in microchimeric cases (1.0+/-0.58) than in nonmicrochimeric ones (1.9+/-0.38). But regarding HLA-DQ and other clinical parameters mentioned above, significant difference was not observed. We propose that there is an association between HLA DR matching, microchimerism and acute graft rejection in our recipients. Our study demonstrates that, with routine immunosuppressive protocols, higher compatibility of HLA-DR antigens facilitates microchimerism induction. Then, development of new stronger immunosuppressive protocols (including conditioning) or augmentation of chimeric state (by donor-specific bone marrow infusion), especially in completely mismatched HLA-DR renal allograft recipients, may be useful for graft acceptance. PMID- 11389711 TI - Long-term (> or =20 yr) status of 14 cadaveric kidney-transplant recipients. AB - The aim of this study was to analyze the status of patients with a successful long-term (> or =20 yr) kidney graft. Nineteen (8.1%) of the 234 recipients who received a cadaveric kidney transplant between 1968 and 1978 in our center are still alive 21.7+/-1.6 yr (mean+/-standard error of the mean) later with a functioning allograft. Function, including measurement of the renal functional reserve (RFR), histological status, and morbidity were evaluated. Fourteen patients agreed to participate in this study. Their current immunosuppressive regimens combined prednisone (P)+azathioprine (AZA) (n=9), P+AZA+cyclosporine (CsA) (n=3) or P+CsA (n=2). Although they described their quality of life as good, 10 patients had mild hypertension, 5 developed 10 malignancies (9 cutaneous), 5 had replicative hepatitis, 8 had osteopenia, and 6 had cataracts, but none had diabetes mellitus. Proteinuria was detected in 6 patients, but was always less than 1 g/d. Mean serum creatinine was 1.28+/-0.28 mg/dL and glomerular filtration rate was 54.5+/-5.3 mL/min/1.73 m2. RFR was present for 4 patients with a mean value of +14.8+/-1.9 mL/min. Their functional status was not correlated with the histological lesions observed in concomitant transplant biopsies. Kidney grafts are able to function well even more than 20 yr post transplantation, with some having a RFR whose significance remains unknown. Morbidity is of minor clinical severity, but could be further reduced with optimized management. Moreover, transplantation is much less costly than hemodialysis. PMID- 11389712 TI - Sequential measurement of human herpesvirus 6 DNA with polymerase chain reaction method in pediatric living-related liver transplantation. AB - BACKGROUND: Human herpesvirus 6 (HHV-6), a causative virus of exanthem subitum, may occasionally present with a severe clinical form in immunosuppressed patients after transplantation. In this study, HHV-6 DNA was sequentially measured with a polymerase chain reaction (PCR) method, a quick and sensitive modality in pediatric living-related liver transplantation (LTx). METHODS: Subjects consisted of 5 post-operative biliary atresia patients undergoing living-related LTx at ages from 8 months to 4 yr. Immunosuppression was performed with Tacrolimus (blood trough level 8-18 within 1 month and 5-10 ng/mL thereafter) and low-dose steroid. Specimens were peripheral blood mononuclear cells (PBMC), plasma, and liver biopsy tissue. The amount of HHV-6 DNA was semiquantified as follows: 1+, 1 10; 2+, 10-100; 3+, 100-1000; 4+, over 1000 copies/105 PBMCs. RESULTS: A total of 69 blood samples and three liver biopsies were provided for the examination. HHV 6 DNA in PBMC was positive in 2 donors and 3 recipients before LTx. Two patients with negative DNA were converted to 3+ at 2-3 wk after LTx and 3 with positive DNA remained 2+ to 3+ throughout the post-LTx period. Only 1 patient developed clinical symptoms, such as fever, liver dysfunction, petechiae, idiopathic thrombocytopenic purpura, and finally bone marrow suppression. HHV-6 DNA in the liver biopsy tissue and plasma in this patient were 4+ and 2+, respectively. CONCLUSION: HHV-6 DNA in PBMC measured by the PCR method may be persistently high in pediatric recipients after living-related LTx. Although HHV-6 DNA in PBMC may be positive in case of evident infection, positivity in PBMC may not be always associated with the clinical symptoms. PMID- 11389713 TI - Echocardiographic ultrasonic tissue characterization in a case of Fabry's disease following renal transplantation. AB - We report a case of Fabry's disease where stabilization of progressive cardiac involvement was recorded in a 29-yr-old Caucasian man, to our knowledge, for the first time by ultrasonic tissue characterization echocardiography after 1 yr of successful renal transplantation. Three echocardiographic evaluations have been made: the first 3 months before, the second 6 months after, and the third 1 yr after kidney transplantation. The myocardial structural damage - evaluated by integrated backscatter index - shows a persistence of the impairment of intrinsic myocardial contractility at septum level, probably due to coexistent hypertensive status, which is able to induce per se alterations of myocardial textural parameters. On the other hand, the cyclic variation index at posterior free wall, which is less dependent on strictly hemodynamic factors than the septum, appears quite normal at the third observation. These data could reflect the improvement of the ultrastuctural myocardial findings in relation to renal transplantation, which could correct not only renal failure but also the enzymatic deficiency by replacement of alpha-galactosidase A through the transplanted kidney. PMID- 11389714 TI - Structural elucidation of a capsular polysaccharide from a clinical isolate of Bacteroides vulgatus from a patient with Crohn's disease. AB - The structure of a capsular polysaccharide (CPS) from a clinical isolate of Bacteroides vulgatus was elucidated. B. vulgatus IMCJ 1204 was isolated from feces of a patient with Crohn's disease. CPS was prepared by phenol/water extraction of the bacterial cells followed by hydrophobic interaction chromatography and then gel filtration chromatography of the extract. The structure of CPS was determined by chemical analysis and NMR spectroscopy including DQF-COSY, TOCSY, ROESY, HSQC-TOCSY, HMQC and HMBC to be a polysaccharide composed of the following repeating unit: -->3)beta-D-Glcp(1- >6)[alpha-D-GalpNAc(1-->2)beta-D-Galp(1-->4)]beta-D-GlcpNAc(1-->3)alpha-D-Galp(1- >4)beta-D-Manp(1-->. PMID- 11389715 TI - Palmitate uptake by neonatal rat myocytes and hepatocytes. Role of extracellular protein. AB - The role of extracellular binding proteins in the rate of [3H]palmitate uptake by neonatal cardiac myocytes and hepatocytes was investigated using a model independent approach. Binding proteins used in this study included alpha1-acid glycoprotein [isoelectric point (pI) approximately 2.7], conalbumin (pI approximately 6.4), lysozyme (pI approximately 11.0), albumin (pI approximately 4.9), and albumin which had been modified to yield proteins with pI values of 3.5, 4.7, 7.5 and 8.6. All uptake studies were conducted at similar unbound ligand fractions. There was a linear relationship between the rate of neonatal hepatocyte [3H]palmitate clearance and protein pI (r2 = 0.98). In contrast, there was an overall poor relationship between neonatal cardiac myocyte [3H]palmitate clearance rate and protein pI (r2 = 0.48). However, the relationship improved when the data on [3H]palmitate-clearance were analyzed using only the modified albumins. The study indicates that an ionic interaction between extracellular proteins and the hepatocyte surface enhances the overall uptake of [3H]palmitate. This interaction may be limited to albumin for neonatal cardiac myocytes. PMID- 11389716 TI - HMG-1 enhances HMG-I/Y binding to an A/T-rich enhancer element from the pea plastocyanin gene. AB - High-mobility-group proteins HMG-1 and HMG-I/Y bind at overlapping sites within the A/T-rich enhancer element of the pea plastocyanin gene. Competition binding experiments revealed that HMG-1 enhanced the binding of HMG-I/Y to a 31-bp region (P31) of the enhancer. Circularization assays showed that HMG-1, but not HMG-I/Y, was able to bend a linear 100-bp DNA containing P31 so that the ends could be ligated. HMG-1, but not HMG-I/Y, showed preferential binding to the circular 100 bp DNA compared with the equivalent linear DNA, indicating that alteration of the conformation of the DNA by HMG-1 was not responsible for enhanced binding of HMG I/Y. Direct interaction of HMG-I/Y and HMG-1 in the absence of DNA was demonstrated by binding of 35S-labeled proteins to immobilized histidine-tagged proteins, and this was due to an interaction of the N-terminal HMG-box-containing region of HMG-1 and the C-terminal AT-hook region of HMG-I/Y. Kinetic analysis using the IAsys biosensor revealed that HMG-1 had an affinity for immobilized HMG I/Y (Kd = 28 nM) similar to that for immobilized P31 DNA. HMG-1-enhanced binding of HMG-I/Y to the enhancer element appears to be mediated by the formation of an HMG-1-HMG-I/Y complex, which binds to DNA with the rapid loss of HMG-1. PMID- 11389717 TI - Sp1 involvement in the 4beta-phorbol 12-myristate 13-acetate (TPA)-mediated increase in resistance to methotrexate in Chinese hamster ovary cells. AB - 4beta-Phorbol 12-myristate 13-acetate (TPA) increases the number of colonies resistant to methotrexate (MTX), mainly by amplification of the dihydrofolate reductase (dhfr) locus. We showed previously that inhibition of protein kinase C (PKC) prevents this resistance. Here, we studied the molecular changes involved in the development of TPA-mediated MTX resistance in Chinese hamster ovary (CHO) cells. TPA incubation increased the expression and activity of DHFR. Because Sp1 controls the dhfr promoter, we determined the effect of TPA on the expression of Sp1 and its binding to DNA. TPA incubation increased Sp1 binding and the levels of Sp1 protein. The latter effect was due to an increase in Sp1 mRNA. Dephosphorylation of nuclear extracts from control or TPA-treated cells reduced the binding of Sp1. Stable transfectants of PKCalpha showed increased Sp1 binding, and when treated with MTX, developed a greater number of resistant colonies than control cells. Seventy-five percent of the isolated colonies showed increased copy number for the dhfr gene. Transient expression of PKCalpha increased DHFR activity. Over-expression of Sp1 increased resistance to MTX, and inhibition of Sp1 binding by mithramycin decreased this resistance. We conclude that one mechanism by which TPA enhances MTX resistance, mainly by gene amplification, is through an increase in Sp1 expression which leads to DHFR activation. PMID- 11389718 TI - Structure of an acidic exopolysaccharide produced by the diazotrophic endophytic bacterium Burkholderia brasiliensis. AB - The structure of an exopolysaccharide (EPS) produced by Burkholderia brasiliensis, a diazotrophic endophytic organism originally isolated from rice roots, has been determined. The bacterium was grown in a synthetic medium, containing mannitol and glutamate, which favours the expression of two anionic EPSs, which were separated by anion-exchange chromatography. The structure of the repeat unit of EPS A, eluted at higher ionic strength, was determined by a combination of methylation analysis, partial hydrolysis, chemical degradations, and NMR spectroscopic studies, and shown to be the linear O-acetylated pentasaccharide: -->4)-alpha-D-Glcp-(1-->2)-alpha-L-Rhap-(1-->4)-alpha-D-GlcpA-(1 ->3)-beta-L-Rhap[2OAc]-(1-->4)-beta-D-Glcp-(1-->. PMID- 11389719 TI - The two-component regulatory system BacRS is associated with bacitracin 'self resistance' of Bacillus licheniformis ATCC 10716. AB - Bacitracin is a peptide antibiotic produced by several Bacillus licheniformis strains that is most active against other Gram-positive microorganisms, but not against the producer strain itself. Recently, heterologous expression of the bacitracin resistance mediating BcrABC transporter in Bacillus subtilis and Escherichia coli was described. In this study we could determine that the transporter encoding bcrABC genes are localized about 3 kb downstream of the 44 kb bacitracin biosynthetic operon bacABC. Between the bac operon and the bcrABC genes two orfs, designated bacR and bacS, were identified. They code for proteins with high homology to regulator and sensor proteins of two-component systems. A disruption mutant of the bacRS genes was constructed. While the mutant displayed no effects on the bacitracin production it exhibited highly increased bacitracin sensitivity compared to the wild-type strain. Western blot analysis of the expression of BcrA, the ATP-binding cassette of the transporter, showed in the wild-type a moderate BcrA induction in late stationary cells that accumulate bacitracin, whereas in the bacRS mutant cells the BcrA expression was constitutive. A comparison of bacitracin stressed and nonstressed wild-type cells in Western blot analysis revealed increasing amounts of BcrA and a decrease in BacR in the stressed cells. From these findings we infer that BacR acts as a negative regulator for controlling the expression of the bcrABC transporter genes. PMID- 11389720 TI - Biosynthesis of terpenoids. 2C-Methyl-D-erythritol 2,4-cyclodiphosphate synthase (IspF) from Plasmodium falciparum. AB - The putative catalytic domain of an open reading frame from Plasmodium falciparum with similarity to the ispF gene of Escherichia coli specifying 2C-methyl-D erythritol 2,4-cyclodiphosphate synthase was expressed in a recombinant E. coli strain. The recombinant protein was purified to homogeneity and was found to catalyze the formation of 2C-methyl-D-erythritol 2,4-cyclodiphosphate from 4 diphosphocytidyl-2C-methyl-D-erythritol 2-phosphate at a rate of 4.3 micromol x mg(-1) x min(-1). At lower rates, the recombinant protein catalyzes the formation of 2-phospho-2C-methyl-D-erythritol 3,4-cyclophosphate from 4-diphosphocytidyl-2C methyl-D-erythritol 2-phosphate and the formation of 2C-methyl-D-erythritol 3,4 cyclophosphate from 4-diphosphocytidyl-2C-methyl-D-erythritol. Divalent metal ions such as magnesium or manganese are required for catalytic activity. The enzyme has a pH optimum at pH 7.0. Recombinant expression of the full-length open reading frame afforded insoluble protein that could not be folded in vitro. The enzyme is a potential target for antimalarial drugs directed at the nonmevalonate pathway of isoprenoid biosynthesis. PMID- 11389721 TI - Purification and cDNA cloning of nitric oxide reductase cytochrome P450nor (CYP55A4) from Trichosporon cutaneum. AB - Cytochrome P450nor is involved in fungal denitrification as nitric oxide (NO) reductase. Although the heme protein has been known to occur in restricted species of fungi that belong to ascomycotina, we have previously suggested that it would also occur in the yeast Trichosporon cutaneum, which is phylogenetically far from those P450nor-producing ascomycetous fungi. Here we isolated and characterized the heme protein from the basidiomycetous yeast T. cutaneum. P450nor of the yeast (TcP450nor) exhibited properties in terms of catalysis, absorption spectrum and molecular mass that are almost identical to those of its counterparts in ascomycetous fungi. We also isolated and sequenced its cDNA. The predicted primary structure of TcP450nor showed high sequence identities (around 65%) to those of other P450nors, indicating that they belong to the same family. TcP450nor protein cofractionated with cytochrome c oxidase by subcellular fractionation and its predicted primary structure contained an extension on its amino terminus that is characteristic of a mitochondrial-targeting signal, indicating that it is a mitochondrial protein like some of the isoforms of other fungi. On the other hand, TcP450nor was unique in that inducers such as nitrate, nitrite, or NO were not required for its production in the cells. The occurrence of P450nor across the subdivisions of eumycota suggests that P450nor and denitrification are distributed more universally among fungi than was previously thought. PMID- 11389722 TI - Cassette mutagenesis of lysine 130 of human glutamate dehydrogenase. An essential residue in catalysis. AB - It has been suggested that reactive lysine residue(s) may play an important role in the catalytic activities of glutamate dehydrogenase (GDH). There are, however, conflicting views as to whether the lysine residues are involved in Schiff's base formation with catalytic intermediates, stabilization of negatively charged groups or the carbonyl group of 2-oxoglutarate during catalysis, or some other function. We have expanded on these speculations by constructing a series of cassette mutations at Lys130, a residue that has been speculated to be responsible for the activity of GDH and the inactivation of GDH by pyridoxal 5' phosphate (PLP). For these studies, a 1557-bp gene that encodes human GDH has been synthesized and inserted into Escherichia coli expression vectors. The mutant enzymes containing Glu, Gly, Met, Ser, or Tyr at position 130, as well as the wild-type human GDH encoded by the synthetic gene, were efficiently expressed as a soluble protein and are indistinguishable from that isolated from human and bovine tissues. Despite an approximately 400-fold decrease in the respective apparent Vmax of the Lys130 mutant enzymes, apparent Km values for NADH and 2 oxoglutarate were almost unchanged, suggesting the direct involvement of Lys130 in catalysis rather than in the binding of coenzyme or substrate. Unlike the wild type GDH, the mutant enzymes were unable to interact with PLP, indicating that Lys130 plays an important role in PLP binding. The results with analogs of PLP suggest that the aldehyde moiety of PLP, but not the phosphate moiety, is required for efficient binding to GDH. PMID- 11389723 TI - Mechanism of nitrite-stimulated catalysis by lactoperoxidase. AB - The reactions of lactoperoxidase (LPO) intermediates compound I, compound II and compound III, with nitrite (NO2(-)) were investigated. Reduction of compound I by NO2(-) was rapid (k2 = 2.3 x 10(7) M(-1) x s(-1); pH = 7.2) and compound II was not an intermediate, indicating that NO2* radicals are not produced when NO2(-) reacts with compound I. The second-order rate constant for the reaction of compound II with NO2(-) at pH = 7.2 was 3.5 x 10(5) M(-1) x s(-1). The reaction of compound III with NO2(-) exhibited saturation behaviour when the observed pseudo first-order rate constants were plotted against NO2(-) concentrations and could be quantitatively explained by the formation of a 1 : 1 ratio compound III/NO2(-) complex. The Km of compound III for NO2(-) was 1.7 x 10(-4) M and the first-order decay constant of the compound III/ NO2(-) complex was 12.5 +/- 0.6 s(-1). The second-order rate constant for the reaction of the complex with NO2(-) was 3.3 x 10(3) M(-1) x s(-1). Rate enhancement by NO2(-) does not require NO2* as a redox intermediate. NO2(-) accelerates the overall rate of catalysis by reducing compound II to the ferric state. With increasing levels of H2O2, there is an increased tendency for the catalytically dead-end intermediate compound III to form. Under these conditions, the 'rescue' reaction of NO2(-) with compound III to form compound II will maintain the peroxidatic cycle of the enzyme. PMID- 11389724 TI - Molecular cloning, characterization and regulation by cadmium of a superoxide dismutase from the ectomycorrhizal fungus Paxillus involutus. AB - The gene encoding a superoxide dismutase (PiSOD) was cloned by suppressive subtractive hybridization from cDNA library of the ectomycorrhizal fungus, Paxillus involutus, grown under cadmium-stress conditions. The encoded protein was presumed to be localized in the peroxisomes because it contained a C-terminal peroxisomal localization peptide (SKL) and lacked an N-terminal mitochondrial transit peptide. Complementation of an Escherichia coli SOD null strain that is unable to grow in the presence of paraquat or cadmium indicated that cloned Pisod encoded a functional superoxide dismutase. Sensitivity of PiSOD activity to H2O2 but not KCN, and sequence homologies to other SODs strongly suggest that it is a manganese-containing superoxide dismutase. Monitoring PiSOD transcript, immunoreactive polypeptide and superoxide dismutase activity following cadmium stress suggests that the principal level of control is post-translational. This is, to our knowledge, the first insight in the characterization of molecular events that take place in an ectomycorrhizal fungus during exposure to heavy metals. PMID- 11389725 TI - Thermal stability of pyrrolidone carboxyl peptidases from the hyperthermophilic Archaeon, Pyrococcus furiosus. AB - The temperature adaptation of pyrrolidone carboxyl peptidase (PCP) from a hyperthermophile, Pyrococcus furiosus (Pf PCP), was characterized in the context of an assembly form of the protein which is a homotetramer at neutral pH. The Pf PCP exhibited maximal catalytic activity at 90-95 degrees C and its activity was higher in the temperature range 30-100 degrees C than its counterpart from the mesophilic Bacillus amyloliquefaciens (BaPCP). Thermal stability was monitored by differential scanning calorimetry (DSC). Two clearly separated peaks appeared on the DSC curves for Pf PCP at alkaline and acidic pH. Using the oxidized Pf PCP and two mutant proteins (Pf C188S and Pf C142/188S), it was found that the peaks on the high and low temperature sides of the DSC curve of Pf PCP were produced by the forms with an intersubunit disulfide bridge between the two subunits and without the bridge, respectively, indicating the stabilization effect of intersubunit disulfide bridges. The denaturation temperature (Td) of Pf PCP with intersubunit disulfide bridges was higher by 53 degrees C at pH 9.0 than that of BaPCP. An analysis of the equilibrium ultracentrifugation patterns showed that the tetrameric Pf C142/188S dissociated into dimers with decreasing pH in the acidic region and became monomer subunits at pH 2.5. The heat denaturation of Pf PCP and its two Cys mutants was highly reversible in the dimeric forms, but completely irreversible in the tetrameric form. The Td of Pf C142/188S decreased as the enzyme became dissociated, but the monomeric form of the protein was still folded at pH 2.5, although BaPCP was completely denatured at acidic pH. These results indicate that subunit interaction plays an important role in stabilizing PCP from P. furiosus in addition to the intrinsic enhanced stability of its monomer. PMID- 11389726 TI - Ets1 is a common element in directing transcription of the alpha and beta genes of human protein kinase CK2. AB - Protein kinase CK2 is a conserved and vital Ser/Thr phosphotransferase with various links to malignant diseases, occurring as a tetramer composed of two catalytically active (CK2alpha and/or CK2alpha') and two regulatory subunits (CK2beta). There is balanced availability of CK2alpha and CK2beta transcripts in proliferating and differentiating cultured cells. Examination of the human CK2beta gene for transcriptionally active regions by systematic deletions and reporter gene assays indicates strong promoter activity at positions -42 to 14 and 12 to 72 containing transcription start sites 1 and 2 of the gene (positions +1 and 33), respectively, an upstream and a downstream enhancer activity at positions -241 to -168 and 123 to 677, respectively, and silencer activity at positions -241 to -261. Of the various transcription factor binding motifs present in those regions, Ets1 and CAAT-related motifs turned out to be of particular importance, Ets1 for promoter activation and CAAT-related motifs for enhancer activation. In addition, there are contributions by Sp1. Most strikingly, the Ets1 region representing two adjoining consensus motifs also occurs with complete identity in the recently characterized promoter of the CK2alpha gene [Krehan, A., Ansuini, H., Bocher, O., Grein, S., Wirkner, U. & Pyerin, W. (2001) J. Biol. Chem. 275, 18327-18336], and affects comparably, when assayed in parallel, the promoters of both CK2 genes, both by motif mutations and by Ets1 overexpression. The data strongly support the hypothesis that Ets1 acts as a common regulatory element of the CK2alpha and CK2beta genes involved in directing coordinate transcription and contributing to the balanced availability of transcripts. PMID- 11389727 TI - Modulation of the catalytic activity of cruzipain, the major cysteine proteinase from Trypanosoma cruzi, by temperature and pH. AB - Cysteine proteinases are relevant to several aspects of the parasite life cycle and of parasite-host relationships. Here, a quantitative investigation of the effect of temperature and pH on the total substrate inhibition of cruzipain, the major papain-like cysteine proteinase from Trypanosoma cruzi, is reported. Values of the apparent catalytic and inhibition parameters Km, Vmax, Vmax/Km, and K(i) for the cruzipain-catalysed hydrolysis of N-alpha-benzyloxycarbonyl-L phenylalanyl-L-arginine-(7-amino-4-methylcoumarin) (Z-Phe-Arg-AMC) and azocasein were determined between 10.0 degrees C and 40.0 degrees C and between pH 4.5 and 8.5. Values of Km were independent of temperature and pH, whereas values of Vmax, Vmax/Km, and K(i) were temperature-dependent and pH-dependent. Over the whole pH range explored, values of logVmax, log(Vmax/Km), and logK(i) increased linearly with respect to T(-1). Values of Vmax and Vmax/Km were affected by the acid-base equilibrium of one temperature-independent ionizing group (i.e. pK(unl)' = pK(lig)' = 5.7 +/- 0.1, at 25.0 degrees C). Moreover, values of K(i) were affected by the alkaline pK shift of one ionizing group of active cruzipain (from pK(unl)" = 5.7 +/- 0.1 to pK(lig)" = 6.1 +/- 0.1, at 25.0 degrees C) upon Z-Phe Arg-AMC binding. Values of logK(unl)', logK(lig)', and logK(lig)" were temperature-independent. Conversely, values of logK(unl)" were linearly dependent on T(-1). As a whole, total substrate inhibition of cruzipain decreased with increasing temperature and pH. These data suggest that both synthetic and protein substrates can bind to the unique active centre of cruzipain either productively or following a binding mode which results in enzyme inhibition. However, allosteric effect(s) cannot be excluded. PMID- 11389728 TI - Expression of placental leucine aminopeptidase/oxytocinase in neuronal cells and its action on neuronal peptides. AB - The placental leucine aminopeptidase (P-LAP)/oxytocinase whose serum level increases with gestation is thought to contribute to the maintenance of normal pregnancy. P-LAP mRNAs are expressed in various tissues other than the placenta. In this study, we identified P-LAP protein in the brain. In contrast with the placenta where a significant portion of P-LAP is released, the enzyme was localized in the membrane fraction in brain and PC12 cells and no soluble form of the enzyme was detected. When PC12 cells were differentiated into neuronal cells by nerve growth factor (NGF), a significant increase in the expression level of P LAP in the cell was observed. As in the case of insulin treatment of 3T3-L1 adipocytes, treatment of PC12 cells with forskolin caused the translocation of the enzyme from intracellular vesicle to the cell surface plasma membrane. In addition, P-LAP was shown to degrade several bioactive neuropeptides such as Met enkephalin and dynorphin A (1-8). These results suggest that P-LAP plays an important role in the regulation of neuronal cell function in the brain. PMID- 11389729 TI - Dimethylsulfoxide promotes K+-independent activity of pyruvate kinase and the acquisition of the active catalytic conformation. AB - Pyruvate kinase requires K+ for maximal activity; the enzyme exhibits 0.02% of maximal activity in its absence [Kayne, F. J. (1971) Arch. Biochem. Biophys. 143, 232-239]. However, pyruvate kinase entrapped in reverse micelles exhibits an important K+-independent activity [Ramirez-Silva, L., Tuena de Gomez-Puyou, M., & Gomez-Puyou, A. (1993) Biochemistry 32, 5332-5338]. It is possible that the amount of water, as well as interactions of the protein with the micelles, can account for this behavior. We therefore explored the solvent effects on the catalytic properties of muscle pyruvate kinase. The enzyme exhibited an activity of 19.4 micromol x min(-1) x mg(-1) in 40% dimethylsulfoxide, compared with 280 and 0.023 micromol x min(1) x mg(-1) observed with and without K+ in water, respectively. pH activity profiles and kinetic constants for the substrates of pyruvate kinase in dimethylsulfoxide without K+ were similar to those in 100% water with K+, and differed from those in water without K+. The spectral center of mass of the emission spectrum of pyruvate kinase in 100% water exhibited a blue shift of 3.5 nm in the presence of Mg(2+), phosphenolpyruvate, and K+, ligands that induce the active conformation of the enzyme. The spectral center of mass of the apoenzyme in 30-40% dimethylsulfoxide coincided with that of the enzyme-Mg(2+)-phosphenolpyruvate-K+ complex in 100% water. The water relaxation rate enhancement factor and binding of phosphenolpyruvate to the pyruvate kinase Mn(2+)-(CH3)4N+ complex in 30-40% dimethylsulfoxide were similar to those of the pyruvate kinase-Mn(2+)-K+ complex in water. The aforementioned results indicate that when muscle pyruvate kinase is without K+, 30-40% dimethylsulfoxide induces its active conformation. PMID- 11389730 TI - Differential actions of p60c-Src and Lck kinases on the Ras regulators p120-GAP and GDP/GTP exchange factor CDC25Mm. AB - It is known that the human Ras GTPase activating protein (GAP) p120-GAP can be phosphorylated by different members of the Src kinase family and recently phosphorylation of the GDP/GTP exchange factor (GEF) CDC25Mm/GRF1 by proteins of the Src kinase family has been revealed in vivo [Kiyono, M., Kaziro, Y. & Satoh, T. (2000) J. Biol. Chem. 275, 5441-5446]. As it still remains unclear how these phosphorylations can influence the Ras pathway we have analyzed the ability of p60c-Src and Lck to phosphorylate these two Ras regulators and have compared the activity of the phosphorylated and unphosphorylated forms. Both kinases were found to phosphorylate full-length or truncated forms of GAP and GEF. The use of the catalytic domain of p60c-Src showed that its SH3/SH2 domains are not required for the interaction and the phosphorylation of both regulators. Remarkably, the phosphorylations by the two kinases were accompanied by different functional effects. The phosphorylation of p120-GAP by p60c-Src inhibited its ability to stimulate the Ha-Ras-GTPase activity, whereas phosphorylation by Lck did not display any effect. A different picture became evident with CDC25Mm; phosphorylation by Lck increased its capacity to stimulate the GDP/GTP exchange on Ha-Ras, whereas its phosphorylation by p60c-Src was ineffective. Our results suggest that phosphorylation by p60c-Src and Lck is a selective process that can modulate the activity of p120-GAP and CDC25Mm towards Ras proteins. PMID- 11389731 TI - Design, synthesis and properties of synthetic chlorophyll proteins. AB - A chemoselective method is described for coupling chlorophyll derivatives with an aldehyde group to synthetic peptides or proteins modified with an aminoxyacetyl group at the epsilon-amino group of a lysine residue. Three template-assembled antiparallel four-helix bundles were synthesized for the ligation of one or two chlorophylls. This was achieved by coupling unprotected peptides to cysteine residues of a cyclic decapeptide by thioether formation. The amphiphilic helices were designed to form a hydrophobic pocket for the chlorophyll derivatives. Chlorophyll derivatives Zn-methyl-pheophorbide b and Zn-methyl-pyropheophorbide d were used. The aldehyde group of these chlorophyll derivatives was ligated to the modified lysine group to form an oxime bond. The peptide-chlorophyll conjugates were characterized by electrospray mass spectrometry, analytical HPLC, and UV/visible spectroscopy. Two four-helix bundle chlorophyll conjugates were further characterized by size-exclusion chromatography, circular dichroism, and resonance Raman spectroscopy. PMID- 11389732 TI - The pro-sequence facilitates folding of human nerve growth factor from Escherichia coli inclusion bodies. AB - Nerve growth factor (beta-NGF), a neurotrophin required for the development and survival of specific neuronal populations, is translated as a prepro-protein in vivo. While the presequence mediates translocation into the endoplasmic reticulum, the function of the pro-peptide is so far unknown. As the pro sequences of several proteins are known to promote folding of the mature part, the renaturation behaviour of recombinant human beta-NGF pro-protein was compared to that of the mature form. Expression of rh-pro-NGF in Escherichia coli led to the formation of inclusion bodies (IBs). The presence of the covalently attached pro-sequence significantly increased the yield and rate of refolding with concomitant disulfide bond formation when compared to the in vitro refolding of mature NGF (rh-NGF). Physicochemical characterization revealed that rh-pro-NGF is a dimer. The pro-peptide could be removed by limited proteolysis with trypsin yielding biologically active, mature rh-NGF. Furthermore, rh-pro-NGF exhibited biological activity in the same concentration range as rh-NGF. PMID- 11389734 TI - Functional and computer modelling studies of haemoglobin from horse. The haemoglobin system of the Sardinian wild dwarf horse. AB - A study was made of the haemoglobin (Hb) system from the Sardinian dwarf horse (Equus caballus jara), one of the last surviving wild horse species in Europe. The oxygen binding properties of the whole haemolysate and of the four different horse Hbs, separated by ion-exchange chromatography, were studied with special regard to the effect of chloride, 2,3-diphosphoglycerate and lactate. Results indicate that no significant functional differences exist between the four Hb components of horse haemolysate. Moreover, the molecular basis of the intrinsically low oxygen affinity and of the weak interaction of horse Hb with 2,3-diphosphoglycerate is discussed in the light of the primary structure of the molecule and of the results of a computer modelling approach. On these bases, it is suggested that the A1 (Thr-->Ser) and A2 (Pro-->Gly) substitutions observed in the beta chains from horse Hb may be responsible for the displacement of the A helix that is known to be a key structural feature of those Hbs that display an altered interaction with 2,3-diphosphoglycerate as compared with human Hb. PMID- 11389733 TI - Allosteric sites of phosphodiesterase-5 (PDE5). A potential role in negative feedback regulation of cGMP signaling in corpus cavernosum. AB - To date, relative cellular levels of cGMP and cGMP-binding proteins have not been considered important in the regulation of smooth muscle or any other tissue. In rabbit penile corpus cavernosum, intracellular cGMP was determined to be 18 +/- 4 nM, whereas the cGMP-binding sites of types Ialpha and Ibeta cGMP-dependent protein kinase (PKG) and cGMP-binding cGMP-specific phosphodiesterase (PDE5) were 58 +/- 14 nM and 188 +/- 6 nM, respectively, as estimated by two different methods for each protein. Thus, total cGMP-binding sites (246 nM) greatly exceed total cGMP. Given this excess of cGMP-binding sites and the high affinities of PKG and PDE5 for cGMP, it is likely that a large portion of intracellular cGMP is associated with these proteins, which could provide a dynamic reservoir for cGMP. Phosphorylation of PDE5 by PKG is known to increase the affinity of PDE5 allosteric sites for cGMP, suggesting the potential for regulation of a reservoir of cGMP bound to this protein. Enhanced binding of cGMP by phosphorylated PDE5 could reduce the amount of cGMP available for activation of PKG, contributing to feedback inhibition of smooth muscle relaxation or other processes. This introduces a new concept for cyclic nucleotide signaling. PMID- 11389736 TI - Paralogous gene analysis reveals a highly enantioselective 1,2-O isopropylideneglycerol caprylate esterase of Bacillus subtilis. AB - Carboxylesterase NP of Bacillus subtilis Thai I-8, characterized in 1992 as a very enantioselective (S)-naproxen esterase, was found to show no enantiopreference towards (S)-1,2-O-isopropylideneglycerol (IPG) esters. The ybfK gene was identified by the B. subtilis genome project as an unknown gene with homology to carboxylesterase NP. The purpose of the present study was to characterize the ybfK gene product in order to determine whether this paralogue of carboxylesterase NP had an altered or enhanced stereospecificity. The ybfK gene was cloned and expressed in B. subtilis using a combination of two strong promoters in a multicopy vector. The enzyme was purified from the cytoplasm of B. subtilis by means of anion exchange and hydrophobic interaction chromatography. The purified YbfK is an enzyme of 296 amino acids and shows an apparent molecular mass of 32 kDa (SDS/PAGE). Comparison of the activities of YbfK and carboxylesterase NP towards caprylate esters of IPG revealed that YbfK produces (S)-IPG with 99.9% enantioselectivity. Therefore, we conclude that we have isolated a paralogue of carboxylesterase NP that can be used for the enantioselective production of (S)-IPG. PMID- 11389735 TI - Cell surface human alpha-L-fucosidase. AB - The acid alpha-L-fucosidase is usually found as a soluble component of lysosomes where fucoglycoconjugates are degraded. In the present investigation, we have demonstrated the existence of a cell surface protein with enzymatic alpha-L fucosidase activity that crossreacts specifically with a rabbit anti-(alpha-L fucosidase) Ig. By different approaches, this alpha-L-fucosidase, which represents 10-20% of the total cellular fucosidase activity, was detected in all the tested human cells (hemopoietic, epithelial, mesenchymal). Two bands of approximately 43-49 kDa were observed, although theoretical data support the possibility of having the same genetic origin that the known 50 to 55-kDa Mr alpha-L-fucosidase. We speculate about an alternative traffic pathway for the plasma membrane alpha-L-fucosidase to work on the rapid turnover of glycoproteins. PMID- 11389737 TI - Purification, cDNA cloning and characterization of the vascular apoptosis inducing protein, HV1, from Trimeresurus flavoviridis. AB - Hemorrhagic snake venom induces apoptosis in vascular endothelial cells (VEC). In previous reports, we described the purification and cDNA cloning from Crotalus atrox of a vascular apoptosis-inducing protein (VAP1) that specifically induces apoptosis in vascular endothelial cells. We report here the purification and cDNA cloning of another vascular apoptosis-inducing protein, HV1, from crude venom of Trimeresurus flavoviridis. The protein, namely HV1, was purified as an inducer of apoptosis in cultured vascular endothelial cells. HV1 was a homodimeric protein with a molecular mass of 110 kDa. HV1 cDNA encoded a protein with 612 amino-acid residues. The amino-acid sequence predicted from the cDNA was highly homologous to VAP1. The amino-acid sequence of HV1 indicated that HV1 belongs to the metalloprotease/disintegrin family, and that it is a multidomain polypeptide with a proprotein domain, a metalloprotease domain, a disintegrin-like domain and a cysteine-rich domain. In the disintegrin-like domain, the sequence DECD, replaces the RGD sequence that has frequently been found in such domains. This replacement also occurs in VAP1. Our results indicate HV1 as the first identified homolog of VAP1. PMID- 11389738 TI - Catalytic selenols couple the redox cycles of metallothionein and glutathione. AB - Co-ordination of zinc to the thiol group of cysteine allows mobilization of zinc through oxidation of its ligand. This molecular property links the binding and release of zinc in metallothionein (MT) to the cellular redox state [Maret W. & Vallee B.L. (1998) Proc. Natl Acad. Sci. USA 95, 3483-3488]. Biological disulfides such as glutathione disulfide (GSSG) oxidize MT with concomitant release of zinc, while glutathione (GSH) reduces the oxidized protein to thionein, which then binds to available zinc. Neither of these two redox processes is very efficient, even at high concentrations of GSSG or GSH. However, the GSH/GSSG redox pair can efficiently couple with the MT/thionein system in the presence of a selenium compound that has the capacity to form a catalytic selenol(ate). This coupling provides a very effective means of modulating oxidation and reduction. Remarkably, selenium compounds catalyze the oxidation of MT even under overall reducing conditions such as those prevailing in the cytosol. In this manner, the binding and release of zinc from zinc-thiolate co ordination sites is linked to redox catalysis by selenium compounds, changes in the glutathione redox state, and the availability of either a zinc donor or a zinc acceptor. The results also suggest that the pharmacological actions of selenium compounds in cancer prevention and other antiviral and anti-inflammatory therapeutic applications, as well as unknown functions of selenium-containing proteins, may relate to coupling between the thiol redox state and the zinc state. PMID- 11389740 TI - Is there a role for magnetic resonance imaging in renal trauma? AB - BACKGROUND: Computed tomography (CT) has been the most informative imaging method in renal trauma. Despite the good sensitivity of magnetic resonance imaging (MRI) to the presence of hematoma, edema and ischemia, MRI has not been widely studied in patients with renal trauma. The present study was initiated to evaluate the role of MRI in patients with renal trauma. METHODS: Between June 1998 and September 1999, CT and MRI were prospectively performed on 12 patients who suffered from renal trauma and the results reviewed. RESULTS: The presence and size of perirenal hematoma could be detected by both CT and MRI. Magnetic resonance imaging could differentiate intrarenal hematoma from perirenal hematoma more accurately, and provided additional information about the hematoma as T1- and T2-weighted MRI were able to determine recent bleeding in the hematoma by regional differences in signal intensity. Magnetic resonance imaging clearly revealed renal fracture with non-viable fragment and detected focal renal laceration that was not detected on CT due to perirenal hematoma associated with renal infarction. However, although MRI had many advantages over CT, it had also major drawbacks, which were that it required longer imaging time and increased the cost. CONCLUSIONS: Magnetic resonance imaging may be useful in renal trauma. However, it is suggested that MRI should be limited to carefully selected patients, such as those with severe renal injury or equivocal findings on CT. PMID- 11389739 TI - Benzalacetone synthase. A novel polyketide synthase that plays a crucial role in the biosynthesis of phenylbutanones in Rheum palmatum. AB - Benzalacetone synthase (BSA) is a novel plant-specific polyketide synthase that catalyzes a one step decarboxylative condensation of 4-coumaroyl-CoA with malonyl CoA to produce the C6-C4 skeleton of phenylbutanoids in higher plants. A cDNA encoding BAS was for the first time cloned and sequenced from rhubarb (Rheum palmatum), a medicinal plant rich in phenylbutanoids including pharmaceutically important phenylbutanone glucoside, lindleyin. The cDNA encoded a 42-kDa protein that shares 60-75% amino-acid sequence identity with other members of the CHS superfamily enzymes. Interestingly, R. palmatum BAS lacks the active-site Phe215 residue (numbering in CHS) which has been proposed to help orient substrates and intermediates during the sequential condensation of 4-coumaroyl-CoA with malonyl CoA in CHS. On the other hand, the catalytic cysteine-histidine dyad (Cys164 His303) in CHS is well conserved in BAS. A recombinant enzyme expressed in Escherichia coli efficiently afforded benzalacetone as a single product from 4 coumaroyl-CoA and malonyl-CoA. Further, in contrast with CHS that showed broad substrate specificity toward aliphatic CoA esters, BAS did not accept hexanoyl CoA, isobutyryl-CoA, isovaleryl-CoA, and acetyl-CoA as a substrate. Finally, besides the phenylbutanones in rhubarb, BAS has been proposed to play a crucial role for the construction of the C6-C4 moiety of a variety of natural products such as medicinally important gingerols in ginger plant. PMID- 11389741 TI - Cyst-associated renal cell carcinoma: clinicopathologic characteristics and evaluation of prognosis in 27 cases. AB - BACKGROUND: No consistent clinicopathologic characteristics of cyst-associated renal cell carcinoma (CRCC) have previously been determined. METHODS: In total, 768 patients with renal cell carcinoma (RCC) underwent radical or partial nephrectomy. Renal cell carcinoma was classified as CRCC in 27 of these patients (3.5%, subdivided into RCC originating in a cyst and cystic RCC), clear-cell RCC in 662 patients (86.2%), chromophobe cell renal carcinoma in 36 patients (4.7%) and papillary RCC in 43 patients (5.6%) according to the criteria of the World Health Organization. RESULTS: The pathologic stage and nuclear grade were usually lower in those with CRCC (low stage/low grade; 89%/96%) or chromophobe cell renal carcinoma (low stage/low grade; 89%/80%) than in those with clear-cell RCC (low stage/low grade; 59%/65%) or papillary RCC (low stage/low grade; 53%/69%). Of the 27 CRCC patients, only 19 (70%) could be diagnosed through preoperative imaging studies. Patients with CRCC showed a favorable prognosis (survival rate: 95% at 1 year, 89.7% at 3 years and 84.4% thereafter) and, especially among the patients with RCC originating in a cyst, no cancer-related death was observed. Comparing the survival among four types of RCC, a favorable outcome was observed in cases of CRCC or chromophobe cell renal carcinoma compared with clear-cell RCC or papillary RCC (clear vs chromophobe: P = 0.002; chromophobe vs papillary: P = 0.019; clear vs cyst-associated: P = 0.001; papillary vs cyst-associated: P = 0.00079). CONCLUSIONS: In cases of CRCC, the disease was usually detected at lower stages and grades and therefore the prognosis was better than in cases of other types of RCC. Preoperative diagnosis of this disease was very difficult, especially in cases of RCC originating in a cyst. PMID- 11389742 TI - Histopathologic analysis of angiogenic factors in localized renal cell carcinoma: the influence of neoadjuvant treatment. AB - BACKGROUND: This study was conducted in order to clarify whether histopathologic analysis of factor thymidine phosphorylase (TP) and Factor VIII could be a useful predictor of postoperative recurrence in localized renal cell carcinoma. Therefore, the relationship between tumor infiltrated lymphocytes (TIL) and both TP and Factor VIII was studied. METHOD: Of the 71 patients who underwent surgery, 54 patients had no neoadjuvant therapy (group 1), 10 patients were preoperatively administered IFN-gamma (group 2), and the remaining seven patients preoperatively received IFN-gamma and transarterial embolization (group 3). Both TP and Factor VIII immunostaining were performed on formalin-fixed, paraffin-embedded archival tissue from 71 renal cell carcinoma specimens, while TIL immunostaining was performed on frozen sections. Positive immunostaining was quantitatively scored by a computer-assisted digital image analysis. For TIL, positive results were semiquantitatively scored. RESULTS: A significant difference in the recurrence free rate was recognized for Groups 1, 2 and 3 (P < 0.05). Therefore, the median TP-positive rate (PR), VIII-PR, number of microvessels and positive mean vascular area levels were investigated, between the recurrence cases (n = 6) and the recurrence-free cases (n = 11). Only the TP-PR levels showed a significant difference among them (P = 0.044). In regards to the neoadjuvant cases, a significant correlation was observed between both VIII-PR and CD4 (r = 0.815) as well as between VIII-PR and CD11b (r = 0.756). CONCLUSION: There was no clear evidence that the neoadjuvant treatment would increase the recurrence-free survival in patients with localized renal cell carcinoma. TP-PR might be a predictor of postoperative recurrence in patients with localized renal cell carcinoma. PMID- 11389743 TI - Urological management of cloacal anomalies. AB - BACKGROUND: A cloacal anomaly results from incomplete urorectal division and is frequently associated with genitourinary abnormalities. Experience of the urological reconstruction of this entity is reported. METHODS: Nine girls with cloacal malformation have been treated at the Osaka Medical Center in the past 9 years. Seven patients were detected by prenatal ultrasonography as having: megacystis (two patients); hydronephrosis (two patients); or hydrometrocolpos (three patients). Two patients underwent prenatal shunt placement between the enlarged bladder and the amniotic space. RESULTS: For neonatal urinary diversion, four patients received cystostomy. Six of seven patients with associated hydrocolpos required intermittent catheterization to decompress an enlarged vagina. Vesicoureteral reflux was detected in 12 ureters of seven patients. Antireflux surgery was indicated in four patients before definitive repair. Definitive reconstruction was performed on eight patients. The posterior sagittal approach was used in all patients. Vaginal reconstruction was done utilizing a perineal skin flap (one patient), a tubularized vaginal flap (three patients), distal rectum (three patients) and total urogenital mobilization (one patient). Postoperatively, urethrovaginal fistula was created in one patient and complete occlusion was seen in one patient. CONCLUSION: The anatomical variety of this entity determines the management options from in utero. It involves not only the creation of three perineal orifices, but also a continent, catheterizable urethra under the stabilization of renal function. Accomplishment of the definitive repair requires the combined expertise of experienced pediatric surgeons and pediatric urologists. PMID- 11389744 TI - Treatment of androgen-independent prostate cancer with dexamethasone: a prospective study in stage D2 patients. AB - PURPOSE: In order to evaluate the efficacy of dexamethasone in the treatment of Japanese men with androgen-independent prostate cancer, a prospective study was conducted using prostate-specific antigen (PSA) as a primary end-point. METHODS: Nineteen Japanese men with stage D2 androgen-independent prostate cancer were registered and treatment was started. After ruling out anti-androgen withdrawal syndrome, they were treated with dexamethasone (1.5 mg daily). Patients were monitored for PSA, symptoms, radiologic response, survival rate, time to disease progression, time to treatment failure and complications. RESULTS: Prostate specific antigen levels decreased in nine patients (50.0%); five (27.8%) showed a 50% or greater decrease and two (11.1%) showed an 80% or greater decrease. For the nine patients, the mean duration of PSA response was 7.3 months and the median duration was 2.1 months (range, 1.2-27.5+). Bone pain, which was noted in 13 patients at study entry, improved in seven patients (53.8%). Of nine patients who had serial radiographic examinations with bone scan, three (33%) showed partial response, two (22%) were stable and four (44%) showed disease progression. Treatment was well tolerated, except for one patient who suffered a severe pulmonary infection. CONCLUSION: Dexamethasone decreased PSA levels and produced subjective symptomatic improvement in the patients with stage D2 androgen-independent prostate cancer. PMID- 11389745 TI - Treatment outcome by risk group after radical prostatectomy in Japanese men. AB - BACKGROUND: North American investigators have suggested the usefulness of risk group stratification based on prostate-specific antigen (PSA), clinical stage and biopsy Gleason score for predicting the biochemical outcome of prostate cancer after radical prostatectomy. There have been no reports of the application of this stratification to early biochemical outcome after radical surgery in Japanese men. METHODS: The study population consisted of 178 men treated with radical retropubic prostatectomy and bilateral pelvic lymph node dissection at Kitasato University Hospital (n = 110) and Kurashiki Central Hospital (n = 68) between October 1992 and May 1999. Pathologic and biochemical outcomes after radical prostatectomy were analyzed based on risk-group stratification. Risk groups were further analyzed according to detailed pathologic findings at biopsy. RESULTS: The median follow-up period for the 178 patients after radical surgery was 41.5 months (range, 2.0--82.0 months; mean, 40.9 months). Fifty-eight patients experienced PSA failure at a median of 8.0 months following surgery (range, 0.0--58.0). Risk-group stratification distinctly defined groups of pathologic findings in the radical prostatectomy specimens. The proportion of patients with PSA failure for low, intermediate and high-risk groups were 9.5%, 23.9% and 56.9%, respectively (P < 0.0001). Use of the number of cores with cancer and maximum cancer length in biopsy cores failed to improve risk stratification for PSA outcome in all risk groups. CONCLUSIONS: Risk-group stratification based on preoperative variables may significantly improve a physician's ability to counsel patients about PSA outcome after radical prostatectomy. Further improvement in risk stratification may call for use of variables other than the pathologic information in biopsy cores. PMID- 11389746 TI - Transperineal 12-core systematic biopsy in the detection of prostate cancer. AB - BACKGROUND: The present study was designed to determine the clinical value of transperineal 12-core systematic prostate biopsy guided by transrectal ultrasonography (TRUS) in the detection of prostate cancer. METHODS: A total of 679 consecutive patients underwent systematic prostate biopsies because of abnormal results on digital rectal examination and/or TRUS and/or an elevated serum prostate-specific antigen level. Systematic six- and 12-core biopsies were taken in 138 patients between April 1994 and February 1995 and in the remaining 541 between March 1995 and February 2000, respectively. Twelve-core biopsy included two samples from the lateral portion of the peripheral zone and four from the anterior portion of the transition zone in addition to the conventional six-core biopsy. RESULTS: In the series overall, systematic biopsy revealed 156 cases of prostate cancer (23.0%). The detection rate increased by 5.2%, although this was statistically not significant, from 18.8% (26/138) by six-core biopsy to 24.0% (130/541) by 12-core biopsy. Out of 130 patients in whom prostate cancer was detected by 12-core biopsy, it was supposed that conventional six-core biopsy would have missed 18 cases (13.8%). CONCLUSIONS: Systematic 12-core biopsy might improve the detection rate for prostate cancer. However, further studies are needed to determine its clinical value in the diagnosis of the disease. PMID- 11389748 TI - Simultaneous bilateral laparoscopic adrenalectomy in ACTH-independent macronodular adrenal hyperplasia. AB - Laparoscopic surgery for urological conditions has now become popular worldwide. The case is reported of a 56-year-old woman who underwent simultaneous bilateral laparoscopic adrenalectomy for adrenocorticotropic hormone-independent macronodular adrenocortical hyperplasia (AIMAH), followed by autotransplantation of resected adrenal gland fragments. Simultaneous laparoscopic adrenalectomies seem feasible for a patient with AIMAH because of its minimally invasive nature. However, autotransplantation of adrenal fragments failed in this patient with AIMAH. PMID- 11389747 TI - Expression of Apg-1, a member of the Hsp110 family, in the human testis and sperm. AB - BACKGROUND: Apg-1 encodes a heat shock protein belonging to the Hsp110 family and is inducible by a 32 degrees C to 39 degrees C heat shock in somatic cells. In mouse testicular germ cells Apg-1 mRNA is constitutively expressed depending on the developmental stage. As human Apg-1 has recently been identified, the expression of Apg-1 in the human testis and sperm was investigated. METHODS: Expression and heat-inducibility of Apg-1 in the human testicular germ cell tumor cell line, NEC8, was analyzed. Using an antimouse Apg-1 antibody, expression of Apg-1 in the human testis and sperm was examined by western blotting after confirmation of the specificity of the antibody. The cells expressing Apg-1 in the testis were further determined by immunohistochemistry. RESULTS: Slight induction of Apg-1 mRNA was detected in NEC8 cells after 32 degrees C to 39 degrees C temperature shift. In the human testis, the antibody specifically recognized Apg-1, which was absent in the testis without germ cells (Sertoli-cell only syndrome) or arrested at spermatogonia. Spermatocytes and spermatids, but not testicular somatic cells, were positively stained with the anti-Apg-1 antibody. By western blot analysis, Apg-1 was detected in the preparation enriched for sperm from normal volunteers and infertile patients, but not from azoospermia patients. CONCLUSION: Apg-1 is developmentally expressed in human testicular germ cells and sperm, suggesting its role in spermatogenesis and fertilization. Identification of substrates for Apg-1 chaperone activity will help elucidate its function. PMID- 11389749 TI - An unusual cause of renal colic: hydatiduria. AB - Hydatid disease of the urinary tract is seen rarely. Hydatiduria may be a finding of renal involvement, but it is a rare cause of renal colic. A case is reported of renal hydatid disease that was diagnosed during the investigation of renal colic. A 38-year-old woman presented with renal colic. She had a history of episodes of renal colic and occasional voiding of grape-like material. No calculus was found in the urinary tract by plain film or ultrasonographic examination. The histopathologic examination of this material revealed daughter cysts that are pathognomonic for hydatid disease. Ultrasound and computed tomography confirmed this diagnosis and right nephrectomy was performed without cyst perforation or any spilling of cyst content. Albendazole, 10 mg/kg per day, was given for 4 weeks (2 weeks preoperatively and 2 weeks postoperatively). PMID- 11389750 TI - Incidentally discovered giant renal arteriovenous malformation. AB - A case is presented of giant renal arteriovenous malformation (AVM). A 61-year old woman was admitted to the National Defense Medical College Hospital for further evaluation of a renal cyst. Doppler ultrasonography and magnetic resonance imaging revealed a giant renal AVM, although the patient had no history nor clinical sign suggesting an AVM. Under the diagnosis of a right renal AVM, the patient underwent AVM resection. PMID- 11389751 TI - Holmium:YAG laser endoureterotomy in the treatment of ureteroenteric stricture following Indiana urinary diversion. AB - A 57-year-old man who had received radical urethrocystectomy and Indiana urinary diversion 6 months earlier was treated for ureteroenteric anastomosis stricture (left side) using a Holmium:YAG laser via antegrade approach. The availability of small (6.9 Fr) flexible ureteroscope, as well as the use of the Holmium:YAG laser has facilitated the ability to precisely incise the stricture under direct endoscopic visualization. The technique is described for laser endoureterotomy in a patient with ureteroenteric stricture following Indiana urinary diversion. PMID- 11389752 TI - Secondary amyloidosis of the bladder causing macroscopic hematuria. AB - Bladder involvement in amyloidosis is unusual. The case of an 80-year-old man with macroscopic hematuria caused by secondary amyloidosis of the bladder is described. Cystoscopic examination revealed only a diffuse edematous area and bleeding. No tumor-like lesions were identified. Transurethral biopsy revealed amyloid deposits. Macroscopic hematuria disappeared spontaneously after cystoscopy and bladder biopsy. The patient has been followed up without treatment and is currently free of symptoms. PMID- 11389753 TI - Limited Wegener's granulomatosis of the epididymis. AB - A case is presented of Wegener's granulomatosis limited to the epididymis in a 32 year-old man. Tests for antineutrophils cytoplasmic antibodies were negative throughout. He showed excellent response to treatment with prednisolone, azathioprine and cotrimoxazole, following recurrence of his disease, after local complete excision. He remains free of disease 40 months after the discontinuation of all his treatment. PMID- 11389754 TI - Auxin distribution and transport during embryonic pattern formation in wheat. AB - Inhibitors of auxin polar transport disrupt normal embryogenesis and thus specific spatial auxin distribution due to auxin movement may be important in establishing embryonic pattern formation in plants. In the present study, the distribution of the photoaffinity labeling agent tritiated 5-azidoindole-3-acetic acid ([3H],5-N3IAA), an analog of indole-3-acetic acid (IAA), was visualized in zygotic wheat (Triticum aestivum L.) embryos grown in vitro and in planta, and used to deduce auxin transport pathways in these embryos. This study provides the first direct evidence that the distribution of auxin, here [3H],5-N3IAA, is heterogeneous and changes during embryo development. In particular, the shift from radial to bilateral symmetry was correlated with a redistribution of [3H],5 N3IAA in the embryo. Furthermore, in bilaterally symmetrical embryos, that is, embryos in the late transition stage or older, the localization of [3H],5-N3IAA was altered by N-1-naphthylphthalamic acid, a specific inhibitor of auxin polar transport. No significant effect was observed in radially symmetrical embryos, that is, globular embryos, or very early transition embryos. Thus, the shift from radial to bilateral symmetry is associated with the onset of active, directed auxin transport involved in auxin redistribution. A change in the distribution of [3H],5-N3IAA was also observed in morphologically abnormal embryos induced on media supplemented with auxin or auxin polar transport inhibitors. By means of a microscale technique, free IAA concentration was measured in in vitro- and in planta-grown embryos and was found to increase during development. Therefore, IAA may be synthesized or released from conjugates in bilaterally symmetrical embryos, although import from surrounding tissues cannot be excluded. PMID- 11389755 TI - Defective cell proliferation in the floral meristem of alloplasmic plants of Nicotiana tabacum leads to abnormal floral organ development and male sterility. AB - Flowers of an alloplasmic male-sterile tobacco line, comprised of the nuclear genome of Nicotiana tabacum and the cytoplasm of Nicotiana repanda, develop short, poorly-pigmented petals and abnormal sterile stamens that often are fused with the carpel wall. The development of flower organ primordia and establishment of boundaries between the different zones in the floral meristem were investigated by performing expression analysis of the tobacco orthologs of the organ identity genes GLO, AG and DEF. These studies support the conclusion that boundary formation was impaired between the organs produced in whorls 3 and 4 resulting in partial fusions between anthers and carpels. According to the investigations cell divisions and floral meristem size in the alloplasmic line were drastically reduced in comparison with the male-fertile tobacco line. The reduction in cell divisions leads to a discrepancy between cell number and cell determination at the stage when petal and stamen primordia should be initiated. At the same stage expression of the homeotic genes was delayed in comparison with the male-fertile line. However, the abnormal organ development was not due to a failure in the spatial expression of the organ identity genes. Instead the aberrant development in the floral organs of whorls 2, 3 and 4 appears to be caused by deficient floral meristem development at an earlier stage. Furthermore, defects in cell proliferation in the floral meristem of the alloplasmic male sterile line correlates with presence of morphologically modified mitochondria. The putative causes of reduced cell number in the floral meristem and the consequences for floral development are discussed. PMID- 11389756 TI - Differential regulation of the NO3- and NH4+ transporter genes AtNrt2.1 and AtAmt1.1 in Arabidopsis: relation with long-distance and local controls by N status of the plant. AB - Regulation of root N uptake by whole-plant signalling of N status was investigated at the molecular level in Arabidopsis thaliana plants through expression analysis of AtNrt2.1 and AtAmt1.1. These two genes encode starvation induced high-affinity NO3- and NH4+ transporters, respectively. Split-root experiments indicate that AtNrt2.1 expression is controlled by shoot-to-root signals of N demand. Together with 15NO3- influx, the steady-state transcript level of this gene is increased in NO3--fed roots in response to N deprivation of another portion of the root system. Thus AtNrt2.1 is the first identified molecular target of the long-distance signalling informing the roots of the whole plant's N status. In contrast, AtAmt1.1 expression is predominantly dependent on the local N status of the roots, as it is mostly stimulated in the portion of the root system directly experiencing N starvation. The same behaviour was found for NH4+ influx, suggesting that the NH4+ uptake system is much less efficient than the NO3- uptake system, to compensate for a spatial restriction of N availability. Other major differences were found between the regulations of AtNrt2.1 and AtAmt1.1 expression. AtNrt2.1 is strongly upregulated by moderate level of N limitation, while AtAmt1.1 transcript level is markedly increased only under severe N deficiency. Unlike AtNrt2.1, AtAmt1.1 expression is not stimulated in a nitrate reductase-deficient mutant after transfer to NO3- as sole N source, indicating that NO3- per se acts as a signal repressing transcription of AtAmt1.1. These results reveal two fundamentally different types of mechanism involved in the feedback regulation of root N acquisition by the N status of the plant. PMID- 11389757 TI - The 5' untranslated region of the At-P5R gene is involved in both transcriptional and post-transcriptional regulation. AB - The steady-state level of transcripts coding for the pyrroline-5-carboxylate reductase of Arabidopsis (At-P5R) increased under salt and heat stress, mainly because of an enhanced mRNA stability. However, the At-P5R protein level was not induced, and its translation was inhibited at initiation stage and probably also at later stages. Replacement of the 5' untranslated region (5'UTR) and beta glucuronidase (gus) fusion analysis revealed that the first 92 bp region of the At-P5R 5'UTR was sufficient to mediate transcript stabilization and translation inhibition during salt and heat stresses. Furthermore, the first 92 bp region of the At-P5R 5'UTR was also involved in transcription efficiency in a promoter dependent manner. The results demonstrated that the stress regulation of At-P5R is complex and involves the 5'UTR which acts at three levels, partly in opposing directions. PMID- 11389758 TI - The male sterile G cytoplasm of wild beet displays modified mitochondrial respiratory complexes. AB - Cytoplasmic male sterility (CMS) in higher plants has been mainly studied in cultivated species. In most cases, pollen abortion is linked to the presence of an additional mitochondrial polypeptide leading to organelle dysfunction in reproductive tissues. In wild beet, both CMS and hermaphrodite plants coexist in natural populations. The G cytoplasm is widely distributed along the Western European coast, and previous genetic studies have demonstrated that this cytoplasm confers male sterility in beet. In the present study, we have identified two mutations of G mitochondrial genes, each of which results in the production of a respiratory chain complex subunit with an altered molecular weight; the NAD9 subunit has a C-terminal extension while the COX2 subunit has a truncated C-terminus. NADH dehydrogenase activity was unchanged in leaves, but cytochrome c oxidase activity was reduced by 50%. Moreover, Western blot analyses revealed that alternative oxidase was more abundant in male sterile G plants than in a fertile control (Nv), suggesting that this alternative pathway might compensate for the cytochrome c oxidase deficiency. Implications of respiratory chain changes and a putative link with CMS are discussed. PMID- 11389759 TI - Arabidopsis IRT2 gene encodes a root-periphery iron transporter. AB - Iron uptake from the soil is a tightly controlled process in plant roots, involving specialized transporters. One such transporter, IRT1, was identified in Arabidopsis thaliana and shown to function as a broad-range metal ion transporter in yeast. Here we report the cloning and characterization of the IRT2 cDNA, a member of the ZIP family of metal transporters, highly similar to IRT1 at the amino-acid level. IRT2 expression in yeast suppresses the growth defect of iron and zinc transport yeast mutants and enhances iron uptake and accumulation. However, unlike IRT1, IRT2 does not transport manganese or cadmium in yeast. IRT2 expression is detected only in roots of A. thaliana plants, and is upregulated by iron deficiency. By fusing the IRT2 promoter to the uidA reporter gene, we show that the IRT2 promoter is mainly active in the external cell layers of the root subapical zone, and therefore provide the first tissue localization of a plant metal transporter. Altogether, these data support a role for the IRT2 transporter in iron and zinc uptake from the soil in response to iron-limited conditions. PMID- 11389760 TI - Synthesis, processing and export of cytoplasmic endo-beta-1,4-xylanase from barley aleurone during germination. AB - We have identified the major endo-beta-1,4-xylanase (XYN-1) in the aleurone of germinating barley grain, and show that it is expressed as a precursor of Mr 61 500 with both N- and C-terminal propeptides. XYN-1 is synthesized as an inactive enzyme in the cytoplasm, and only becomes active at a late stage of germination when the aleurone ceases to secrete hydrolases. A series of processing steps, mediated in part by aleurone cysteine endoproteases, yields a mature active enzyme of Mr 34 000. Processing and extracellular release of the mature enzyme coincide with the programmed cell death (PCD)-regulated disintegration of aleurone cells. We discuss the significance of delayed aleurone cell-wall degradation by endoxylanases in relation to the secretory capacity of the aleurone, and propose a novel role for aleurone PCD in facilitating the export of hydrolases. PMID- 11389761 TI - Cloning and characterization of irregular xylem4 (irx4): a severely lignin deficient mutant of Arabidopsis. AB - A severe lignin mutant, irx4, has been identified in Arabidopsis thaliana as a result of its collapsed xylem phenotype. In contrast to previously described irx mutants, irx4 plants have 50% less lignin than wild-type plants, whilst the cellulose and hemicellulose content remained unchanged. These alterations in the composition of irx4 secondary cell walls had a dramatic effect on the morphology and architecture of the walls, which expand to fill most of the cell, and also on the physical properties of irx4 stems. Further analysis indicated that the irx4 mutation occurred in a cinnamoyl-CoA reductase (CCR) gene within a highly conserved intron splice site sequence of intron 2. As a result, CCR mRNA transcripts were incorrectly spliced. Transgenic plants expressing an IRX3 promoter-CCR cDNA construct were used to generate a series of plants with varying degrees of lignin content in order to assess the role of lignin content in determining the physical properties of Arabidopsis stems. PMID- 11389762 TI - A highly specific pathogen-responsive promoter element from the immediate-early activated CMPG1 gene in Petroselinum crispum. AB - Within the complex signalling network from pathogen-derived elicitor perception to defense-related gene activation, some immediate-early responding genes may have pivotal roles in downstream transcriptional regulation. We have identified the parsley (Petroselinum crispum) ELI17 gene as a particularly fast-responding gene possessing a new type of W box-containing, elicitor-responsive promoter element, E17. Highly selective E17-mediated reporter gene expression at pathogen infection sites in transgenic Arabidopsis thaliana plants demonstrated the potential of this promoter element for designing new strategies in resistance breeding as well as for further analysis of the early components of defense related gene activation mechanisms. The protein encoded by the ELI17 gene exhibits various structural characteristics of established transcription factors and is designated as a CMPG protein according to the first four strictly conserved amino acids defining a newly emerging class of plant-specific proteins. PMID- 11389763 TI - Identification of a MADS-box gene, FLOWERING LOCUS M, that represses flowering. AB - The timing of flowering is important for the reproductive success of plants. Here we describe the identification and characterization of a new MADS-box gene, FLOWERING LOCUS M (FLM), which is involved in the transition from vegetative to reproductive development. FLM is similar in amino-acid sequence to FLC, another MADS-box gene involved in flowering-time control. flm mutants are early flowering in both inductive and non-inductive photoperiods, and flowering time is sensitive to FLM dosage. FLM overexpression produces late-flowering plants. Thus FLM acts as an inhibitor of flowering. FLM is expressed in areas of cell division such as root and shoot apical regions and leaf primordia. PMID- 11389764 TI - Mitochondrial inheritance in budding yeast. AB - During the past decade significant advances were made toward understanding the mechanism of mitochondrial inheritance in the yeast Saccharomyces cerevisiae. A combination of genetics, cell-free assays and microscopy has led to the discovery of a great number of components. These fall into three major categories: cytoskeletal elements, mitochondrial membrane components and regulatory proteins. These proteins mediate activities, including movement of mitochondria from mother cells to buds, segregation of mitochondria and mitochondrial DNA, and equal distribution of the organelle between mother cells and buds during yeast cell division. PMID- 11389765 TI - Signaling on the endocytic pathway. AB - Ligand binding to receptor tyrosine kinases and G-protein-coupled receptors initiates signal transduction events and induces receptor endocytosis via clathrin-coated pits and vesicles. While receptor-mediated endocytosis has been traditionally considered an effective mechanism to attenuate ligand-activated responses, more recent studies demonstrate that signaling continues on the endocytic pathway. In fact, certain signaling events, such as the activation of the extracellular signal-regulated kinases, appear to require endocytosis. Protein components of signal transduction cascades can assemble at clathrin coated pits and remain associated with endocytic vesicles following their dynamin dependent release from the plasma membrane. Thus, endocytic vesicles can function as a signaling compartment distinct from the plasma membrane. These observations demonstrate that endocytosis plays an important role in the activation and propagation of signaling pathways. PMID- 11389766 TI - Nuclear relocation of normal huntingtin. AB - In Huntington's Disease (HD), the huntingtin protein (Htt) includes an expanded polyglutamine domain. Since mutant Htt concentrates in the nucleus of affected neurons, we have inquired whether normal Htt (Q16--23) is also able to access the nucleus. We observe that a major pool of normal full-length Htt of HeLa cells is anchored to endosomes and also detect RNase-sensitive nuclear foci which include a 70-kDa N-terminal Htt fragment. Agents which damage DNA trigger caspase-3 dependent cleavage of Htt and dramatically relocate the 70 kDa fragment to the nucleoplasm. Considering that polyglutamine tracts stimulate caspase activation, mutant Htt is therefore poised to enter the nucleus. These considerations help rationalize the nuclear accumulation of Htt which is characteristic of HD and provide a first example of involvement of caspase cleavage in release of membrane bound proteins which subsequently enter the nucleus. PMID- 11389767 TI - Endocytosis of NBD-sphingolipids in neurons: exclusion from degradative compartments and transport to the Golgi complex. AB - Sphingolipids are abundant constituents of neuronal membranes that have been implicated in intracellular signaling, neurite outgrowth and differentiation. Differential localization and trafficking of lipids to membrane domains contribute to the specialized functions. In non-neuronal cultured cell lines, plasma membrane short-chain sphingomyelin and glucosylceramide are recycled via endosomes or sorted to degradative compartments. However, depending on cell type and lipid membrane composition, short-chain glucosylceramide can also be diverted to the Golgi complex. Here, we show that NBD-labeled glucosylceramide and sphingomyelin are transported from the plasma membrane to the Golgi complex in cultured rat hippocampal neurons irrespective of the stage of neuronal differentiation. Golgi complex localization was confirmed by colocalization and Golgi disruption studies, and importantly did not result from conversion of NBD glucosylceramide or NBD-sphingomyelin to NBD-ceramide. Double-labeling experiments with transferrin or wheat-germ agglutinin showed that NBD sphingolipids are first internalized to early/recycling endosomes, and subsequently transported to the Golgi complex. The internalization of these two sphingolipid analogs was energy and temperature dependent, and their intracellular transport was insensitive to the NBD fluorescence quencher sodium dithionite. These results indicate that vesicles mediate the transport of internalized NBD-glucosylceramide and NBD-sphingomyelin to the Golgi complex. PMID- 11389768 TI - AP-3 adaptor functions in targeting P-selectin to secretory granules in endothelial cells. AB - P-selectin, a cell adhesion protein participating in the early stages of inflammation, contains multiple sorting signals that regulate its cell surface expression. Targeting to secretory granules regulates delivery of P-selectin to the cell surface. Internalization followed by sorting from early to late endosomes mediates rapid removal of P-selectin from the surface. We show here that the P-selectin cytoplasmic domain bound AP-2 and AP-3 adaptor complexes in vitro. The amino acid substitution L768A, which abolishes endosomal sorting and impairs granule targeting of P-selectin, reduced binding of AP-3 adaptors but not AP-2 adaptors. Turnover of P-selectin was 2.4-fold faster than turnover of transferrin receptor in AP-3-deficient mocha fibroblasts, similar to turnover of these two proteins in AP-3-competent cells, demonstrating that AP-3 function is not required for endosomal sorting. However, sorting P-selectin to secretory granules was defective in endothelial cells from AP-3-deficient pearl mice, demonstrating a role for AP-3 adaptors in granule assembly in endothelial cells. P-selectin sorting to platelet alpha-granules was normal in pearl mice, consistent with earlier evidence that granule targeting of P-selectin is mechanistically distinct in endothelial cells and platelets. These observations establish that AP-3 adaptor functions in assembly of conventional secretory granules, in addition to lysosomes and the 'lysosome-like' secretory granules of platelets and melanocytes. PMID- 11389769 TI - The actin cytoskeleton is required for the trafficking of the B cell antigen receptor to the late endosomes. AB - The B cell antigen receptor (BCR) plays two central roles in B cell activation: to internalize antigens for processing and presentation, and to initiate signal transduction cascades that both promote B cells to enter the cell cycle and facilitate antigen processing by accelerating antigen transport. An early event in B cell activation is the association of BCR with the actin cytoskeleton, and an increase in cellular F-actin. Current evidence indicates that the organization of actin filaments changes in response to BCR-signaling, making actin filaments good candidates for regulation of BCR-antigen targeting. Here, we have analyzed the role of actin filaments in BCR-mediated antigen transport, using actin filament-disrupting reagents, cytochalasin D and latrunculin B, and an actin filament-stabilizing reagent, jasplakinolide. Perturbing actin filaments, either by disrupting or stabilizing them, blocked the movement of BCR from the plasma membrane to late endosomes/lysosomes. Cytochalasin D-treatment dramatically reduced the rate of internalization of BCR, and blocked the movement of the BCR from early endosomes to late endosomes/lysosomes, without affecting BCR signaling. Thus, BCR-trafficking requires functional actin filaments for both internalization and movement to late endosomes/lysosomes, defining critical control points in BCR-antigen targeting. PMID- 11389770 TI - Serine/threonine protein phosphatase 5 (PP5) participates in the regulation of glucocorticoid receptor nucleocytoplasmic shuttling. AB - BACKGROUND: In most cells glucocorticoid receptors (GR) reside predominantly in the cytoplasm. Upon hormone binding, the GR translocates into the nucleus, where the hormone-activated GR-complex regulates the transcription of GR-responsive genes. Serine/threonine protein phosphatase type 5 (PP5) associates with the GR heat-shock protein-90 complex, and the suppression of PP5 expression with ISIS 15534 stimulates the activity of GR-responsive reporter plasmids, without affecting the binding of hormone to the GR. RESULTS: To further characterize the mechanism by which PP5 affects GR-induced gene expression, we employed immunofluorescence microscopy to track the movement of a GR-green fluorescent fusion protein (GR-GFP) that retained hormone binding, nuclear translocation activity and specific DNA binding activity, but is incapable of transactivation. In the absence of glucocorticoids, GR-GFP localized mainly in the cytoplasm. Treatment with dexamethasone results in the efficient translocation of GR-GFPs into the nucleus. The nuclear accumulation of GR-GFP, without the addition of glucocorticoids, was also observed when the expression of PP5 was suppressed by treatment with ISIS 15534. In contrast, ISIS 15534 treatment had no apparent effect on calcium induced nuclear translocation of NFAT-GFP. CONCLUSION: These studies suggest that PP5 participates in the regulation of glucocorticoid receptor nucleocytoplasmic shuttling, and that the GR-induced transcriptional activity observed when the expression of PP5 is suppressed by treatment with ISIS 15534 results from the nuclear accumulation of GR in a form that is capable of binding DNA yet still requires agonist to elicit maximal transcriptional activation. PMID- 11389771 TI - A glutamine-amidotransferase-like protein modulates FixT anti-kinase activity in Sinorhizobium meliloti. AB - BACKGROUND: Nitrogen fixation gene expression in Sinorhizobium meliloti, the alfalfa symbiont, depends on a cascade of regulation that involves both positive and negative control. On top of the cascade, the two-component regulatory system FixLJ is activated under the microoxic conditions of the nodule. In addition, activity of the FixLJ system is inhibited by a specific anti-kinase protein, FixT. The physiological significance of this negative regulation by FixT was so far unknown. RESULTS: We have isolated by random Tn5 mutagenesis a S. meliloti mutant strain that escapes repression by FixT. Complementation test and DNA analysis revealed that inactivation of an asparagine synthetase-like gene was responsible for the phenotype of the mutant. This gene, that was named asnO, encodes a protein homologous to glutamine-dependent asparagine synthetases. The asnO gene did not appear to affect asparagine biosynthesis and may instead serve a regulatory function in S. meliloti. We provide evidence that asnO is active during symbiosis. CONCLUSIONS: Isolation of the asnO mutant argues for the existence of a physiological regulation associated with fixT and makes it unlikely that fixT serves a mere homeostatic function in S. meliloti. Our data suggest that asnO might control activity of the FixT protein, in a way that remains to be elucidated. A proposed role for asnO might be to couple nitrogen fixation gene expression in S. meliloti to the nitrogen needs of the cells. PMID- 11389772 TI - The regulation of CD5 expression in murine T cells. AB - BACKGROUND: CD5 is a pan-T cell surface marker that is also present on a subset of B cells, B-1a cells. Functional and developmental subsets of T cells express characteristic CD5 levels that vary over roughly a 30-fold range. Previous investigators have cloned a 1.7 Kb fragment containing the CD5 promoter and showed that it can confer similar lymphocyte-specific expression pattern as observed for endogenous CD5 expression. RESULTS: We further characterize the CD5 promoter and identify minimal and regulatory regions on the CD5 promoter. Using a luciferase reporter system, we show that a 43 bp region on the CD5 promoter regulates CD5 expression in resting mouse thymoma EL4 T cells and that an Ets binding site within the 43 bp region mediates the CD5 expression. In addition, we show that Ets-1, a member of the Ets family of transcription factors, recognizes the Ets binding site in the electrophoretic mobility shift assay (EMSA). This Ets binding site is directly responsible for the increase in reporter activity when co-transfected with increasing amounts of Ets-1 expression plasmid.We also identify two additional evolutionarily-conserved regions in the CD5 promoter (CD5X and CD5Y) and demonstrate the respective roles of the each region in the regulation of CD5 transcription. CONCLUSION: Our studies define a minimal and regulatory promoter for CD5 and show that the CD5 expression level in T cells is at least partially dependent on the level of Ets-1 protein. Based on the findings in this report, we propose a model of CD5 transcriptional regulation in T cells. PMID- 11389774 TI - CMV infection of liver transplant recipients: comparison of antigenemia and molecular biology assays. AB - BACKGROUND: CMV is a major clinical problem in transplant recipients. Thus, it is important to use sensitive and specific diagnostic techniques to rapidly and accurately detect CMV infection and identify patients at risk of developing CMV disease. In the present study, CMV infection after liver transplantation was monitored retrospectively by two molecular biology assays - a quantitative PCR assay and a qualitative NASBA assay. The results were compared with those obtained by prospective pp65 antigenemia determinations. MATERIALS AND METHODS: 87 consecutive samples from 10 liver transplanted patients were tested for CMV by pp65 antigenemia, and CMV monitor and NASBA pp67 mRNA assay. RESULTS: CMV infection was detected in all patients by antigenemia and CMV monitor, whereas NASBA assay identified only 8/10 patients with viremia. Furthermore, CMV infection was never detected earlier by molecular biology assays than by antigenemia. Only 5/10 patients with CMV infection developed CMV disease. Using a cut off value of 8 cells/50,000, antigenemia was found to be the assay that better identified patients at risk of developing CMV disease. However, the kinetics of the onset of infection detected by NASBA and CMV monitor seemed to have better identified patients at risk of developing CMV disease. Furthermore, before onset of disease, CMV pp67 mRNA was found to have similar or better negative and positive predictive values for the development of CMV disease. CONCLUSIONS: The present data, suggests that the concomitant use of antigenemia and pp67 mRNA assay gives the best identification of patients at risk of developing CMV disease. PMID- 11389775 TI - Cytochrome P450 2E1 polymorphism and nasopharyngeal carcinoma development in Thailand: a correlative study. AB - BACKGROUND: Nasopharyngeal carcinoma (NPC) is a rare tumor in most parts of the world but occurs at relatively high frequency among people of Chinese descent. The cytochrome P450 2E1 enzyme (CYP2E1) is responsible for the metabolic activation of nitrosamines, and has been shown to be a susceptibility gene for NPC development in Taiwan [RR = 2.6; 95%CI = 1.2-5.7]. Since there has been only one report of this link, it was decided to investigate the susceptibility of CYP2E1 to NPC development in other populations. Therefore, the correlation between the RsaI polymorphism of this gene and NPC was studied in-patients including Thai and Chinese in Thailand. The present study comprised 217 cases diagnosed with NPC and 297 healthy controls. RESULTS: Similar to the result found in Taiwanese, a homozygous uncut genotype demonstrated a higher relative risk both when all cases were analyzed [RR = 2.19; 95%CI = 0.62-8.68] or individual racial groups, Thai [RR = 1.51; 95%CI = 0.08-90.06] or Chinese [RR = 1.99; 95%CI = 0.39-10.87]. The ethnicity-adjusted odds ratio is 2.39 with 95%CI, 0.72-7.89. CONCLUSIONS: Though our finding was not statistically significant due to the moderate sample size of the study, similarity to the study in Taiwan with only a slight loss in precision was demonstrated. The higher RR found for the same genotype in distinct populations confirmed that CYP2E1 is one of several NPC susceptibility genes and that the RsaI minus variant is one mutation that affects phenotype. PMID- 11389773 TI - Expression of ionotropic glutamate receptors in the retina of the rdta transgenic mouse. AB - BACKGROUND: The expression of retinal CaMKII is up-regulated in the retina of the rdta mouse in which rod photoreceptors are genetically ablated. As ionotropic glutamate receptors are known substrates of CAMKII, this study set out to determine if the protein levels of ionotropic glutamate receptors in the rdta mouse retina are also affected. RESULTS: The NMDA receptor subunits (NR1, NR2A/B) and the GluR1; AMPA receptor subunit (GluR1) were examined in immunolabeled western blots. The results demonstrate that the amounts of NR1 and NR2A/B receptor subunits are significantly increased in crude synaptic membrane fractions isolated from retinae of the rdta mice when compared to their normal, littermate controls. The GluR1 receptor subunit and its phosphorylation are simultaneously increased in retinae of the rdta mice. CONCLUSIONS: These data indicate that the NMDA receptors and AMPA (GluR1) receptors are altered in the retinae of rdta mice that lack rod photoreceptors. Because the rods are lost at an early stage in development, it is likely that these results are indicative of synaptic reorganization in the retina. PMID- 11389776 TI - Antenatal screening and its possible meaning from unborn baby's perspective. AB - In recent decades antenatal screening has become one of the most routine procedure of pregnancy-follow up and the subject of hot debate in bioethics circles. In this paper the rationale behind doing antenatal screening and the actual and potential problems that it may cause will be discussed. The paper will examine the issue from the point of view of parents, health care professionals and, most importantly, the child-to-be. It will show how unthoughtfully antenatal screening is performed and how pregnancy is treated almost as a disease just since the emergence of antenatal screening. Genetic screening and ethical problems caused by the procedure will also be addressed and I will suggest that screening is more to do with the interests of others rather than those of the child-to be. PMID- 11389777 TI - In memoriam: Trudy L. Bush. PMID- 11389780 TI - News from the Society for Women's Health Research: women and tobacco use. PMID- 11389781 TI - The experts discuss irritable bowel syndrome. Interview by Jodi Godfrey Meisler. PMID- 11389782 TI - Nutrition communique: reach out to teens. PMID- 11389783 TI - Lupus: why women? AB - Estrogens are believed to play a role in the etiology of both human and murine systemic lupus erythematosus (lupus, SLE), presumably through the agency of their cellular receptor proteins. There is now considerable interest in the molecular mechanism of action of estrogens in immune tissues, particularly with regard to autoimmune disorders, which are generally more prevalent in women. In this laboratory, an attempt is being made to characterize estrogen receptors in murine models of SLE, namely NZB/W and MRL/MP-lpr/lpr mice, and to try to relate this to estrogen receptor function in vivo. It is hypothesized that estradiol (E(2)), through its receptors, mediates the progression of murine SLE and that in autoimmune disease, the estrogen receptor is functionally or structurally changed, or both. Initial studies suggest there are differences in estrogen receptors between BALB/c mice, which do not get autoimmune disease, and two strains that do, MRL/MP-lpr/lpr and NZB/W mice. There is evidence that in at least one model of SLE, the normal regulation of estrogen action by progesterone may be impaired. In several laboratories, attempts are being made to relate estrogen action to immune function and to autoimmune diseases. The study of estrogen action on the immune system may lead to the development of treatments that attenuate the immunostimulant effects of E(2) in autoimmune diseases such as SLE. PMID- 11389784 TI - The use of hormone replacement therapy in postmenopausal women with type 2 diabetes. AB - In healthy postmenopausal women, estrogen or hormone replacement therapy (ERT or HRT) can alleviate menopausal symptoms and prevent osteoporosis and may also protect against cardiovascular disease (CVD). In addition to improving lipid metabolism, there are reports that estrogen also improves parameters regulating carbohydrate metabolism, including insulin resistance, in healthy women. Therefore, it is likely that ERT or HRT would also benefit women with type 2 diabetes, not only in relieving menopausal symptoms but also in improving the metabolic abnormalities associated with diabetes and in preventing cardiovascular disease. PMID- 11389785 TI - A cardiologist's view of hormone replacement therapy and alternatives for cardioprotection. PMID- 11389786 TI - Gender equity in undergraduate medical education: a status report. AB - This status report summarizes recent data on and studies of women's experiences as medical students. Women medical students in the United States now number over 29,000--44% of enrollees. Despite large increases in the numbers of women students, harassment and gender stereotyping continue to detract from their education and opportunities. Moreover, specialty choices have remained remarkably stable, with comparatively few women entering surgery and most subspecialties. Because equal opportunity has not yet been achieved, medical schools need to monitor the experiences of their trainees and to target interventions where problems still exist in order to ensure that progress toward gender equity continues. PMID- 11389787 TI - Advancing women and closing the leadership gap: the Executive Leadership in Academic Medicine (ELAM) program experience. AB - Women are persistently underrepresented in the higher levels of academic administration despite the fact that they have been entering the medical profession in increasing numbers for at least 20 years and now make up a large proportion of the medical student body and fill a similar proportion of entry level positions in medical schools. Although there are no easy remedies for gender inequities in medical schools, strategies have been proposed and implemented both within academic institutions and more broadly to achieve and sustain the advancement of women faculty to senior level positions. Substantial, sustained efforts to increase programs and activities addressing the major obstacles to advancement of women must be put in place so that the contributions of women can be fully realized and their skills fittingly applied in meeting the medical education and healthcare needs of all people in the 21st century. PMID- 11389788 TI - Physician gender and hormone replacement therapy discussion. AB - Hormone replacement therapy (HRT) is a major issue confronting millions of women today, and general internal medicine and family practice physicians are an important source of information and counseling on this issue. Previous studies have suggested that HRT discussion and prescribing are more frequent with female than with male physicians, but others have suggested age and practice setting may be the reason for observed differences. We attempted to determine if physician gender influenced HRT discussion frequency between physicians and their patients using cross-sectional, secondary analysis of data collected at general internal medicine and family practice clinics at the University of New Mexico Health Sciences Center. Twenty-seven family practice and internal medicine resident physicians (15 female, 12 male) participated. There were 127 female patients age 45 and older. Audio-taped observations of patient-physician visits collected during 1995 for a study on patient-physician communication and patient satisfaction were used. There was less frequent discussion with female than male physicians (OR = 0.42, p = 0.0014). HRT was discussed during 51 visits. Patients initiated HRT discussion in 39.2% of visits in which it was discussed. Patients with diabetes were less likely to discuss HRT (OR = 0.25, p = 0.0122). Increasing year of physician residency was associated with decreased discussion of HRT (OR = 0.51, p < 0.0001). In this health sciences center, with physicians similar in age and training, male physicians discussed HRT significantly more often than did female physicians. PMID- 11389789 TI - Long-term fracture risk among infertile women: a population-based cohort study. AB - Nulliparity has been linked to bone loss and fractures, but the contribution of infertility is unclear. The purpose of this study was to assess the long-term risk of fractures among infertile women. In a population-based retrospective cohort study, all 658 Olmsted County, Minnesota, women with infertility (failure to conceive after 1 year despite intercourse without contraception) first diagnosed at Mayo Clinic in 1935-1964 were followed for fractures. Risk was assessed by comparing new fractures of each type to the number expected from sex specific and age-specific fracture rates in the general population (standardized incidence ratios [SIR]). During 18,130 person-years of follow-up, 184 women experienced at least one fracture when 291 would have been expected on the basis of fracture incidence rates in the general population (SIR 0.6, 95% CI 0.5-0.7). There was no increase in proximal femur fractures (SIR 1.0, 95% CI 0.6-1.6) and a statistically significant decrease in the risk of distal forearm fractures (SIR 0.7, 95% CI 0.5-0.97), two of the three sites traditionally associated with osteoporosis. By contrast, there was a significant increase in subsequent vertebral fractures (SIR 1.9, 95% CI 1.4-2.4) that was consistent across divergent causes of infertility and reported menstrual patterns. Although an apparent increase in the risk of vertebral fractures requires further investigation, we saw no indication of an increase in limb fractures, suggesting that infertility does not have long-term adverse skeletal effects like those reported for athletes and dieters with irregular menses. PMID- 11389791 TI - Clinical implications of elevated lipoprotein(a). PMID- 11389792 TI - Constraint-induced movement therapy to enhance recovery after stroke. AB - A therapeutic approach to rehabilitation of movement after stroke, termed constraint-induced (CI) movement therapy, has been derived from basic research with monkeys given somatosensory deafferentation. CI therapy consists of a family of therapies; their common element is that they induce persons with stroke to greatly increase the use of a more-affected upper extremity (UE) for many hours a day over a 2- to 3-week period. These therapies have significantly improved quality of movement and substantially increased amount of use of a more-affected UE in the activities of daily living in life situations. A number of neuroimaging and transcranial magnetic stimulation studies have shown that the massed practice of CI therapy produces a massive use-dependent cortical reorganization that increases the area of cortex involved in the innervation of movement of the more affected UE. The intensity and schedule of delivery of this very efficacious therapy is quite different from that of more traditional physical rehabilitation approaches. As a result, to be clinically applicable, the CI therapy approach to rehabilitation will likely require a paradigm shift in the delivery of physical rehabilitation services. PMID- 11389793 TI - Body weight-supported treadmill training after stroke. AB - Gait rehabilitation is a major aspect of neurologic rehabilitation. This review focuses on locomotor therapy by treadmill stimulation with partial body weight support (BWS), which has become a very promising treatment concept over the past few years. It enables severely affected patients to follow modern aspects of motor learning, favoring a task-specific approach. Initially two therapists assist the movement, placing the paretic limbs and controlling the trunk movements. As compared with overground walking, patients walked more symmetrically, less spastically, and more efficiently on the treadmill with BWS. Several clinical controlled studies have shown its potential in patients after stroke, who regained walking ability faster in the acute or in the chronic stage. Controlled multicenter trials comparing locomotor and conventional therapy will be the next step. Also, the use of BWS during overground walking could be incorporated into the locomotor treatment program of less affected stroke patients. An electromechanical gait trainer relieving the strenuous effort of the therapists and controlling the trunk in a phase-dependent manner is a new technical alternative for severe stroke patients. PMID- 11389794 TI - Botulinum toxin for the management of muscle overactivity and spasticity after stroke. AB - Stroke is a major cause of disability involving the arm and leg. This disability results from the upper motoneuron syndrome (UMN) evident after stroke. It is commonly associated with spasticity and muscle overactivity, which can lead to abnormal limb posturing that interferes with active and passive function. The origin of limb deformity in patients with UMN is based on the concept of unbalanced agonist and antagonist muscle forces acting across joints. In the past decade, botulinum toxin A (BTX-A) a new medication that modifies muscle force and, hence, can treat muscle imbalance, has become available and has renewed interest in the management of muscle overactivity and spasticity after stroke. A reduction in muscle tone, painful spasms, and improved functionality can be obtained. Research and clinical reports support the concept that chemodenervation with BTX-A is an excellent intervention for treating focal muscle overactivity and spasticity secondary to stroke. Many muscles differing in size, shape, and location have been injected, and clinical effectiveness is particularly notable in elbow flexors, ankle plantar flexors, and smaller limb muscles, such as intrinsics of the hand and wrist. Smaller muscles are readily accessible for injection and require smaller amounts of toxin. PMID- 11389795 TI - Medical therapy for acute ischemic stroke. AB - Over the past decade, there has been an explosion in data related to the treatment of patients with acute ischemic stroke. Thrombolytic therapy with intravenous tissue plasminogen activator has revolutionized the approach to stroke treatment. Intra-arterial administration of thrombolytic agents is also being investigated and is now being used on a compassionate basis. Medical management can have a large impact on stroke-related outcomes, even in patients who do not receive thrombolytic therapy. PMID- 11389796 TI - Post-stroke depression: an update. AB - Those caring for stroke victims should be aware of new developments in our understanding of depression following stroke, its diagnosis, prevalence, pathophysiology, clinical features, and treatment. Appropriate diagnosis and treatment will improve quality of life, self-care independence, and mortality. PMID- 11389797 TI - Controversies about tissue plasminogen activator: extending the window of therapy. AB - The management of stroke has undergone significant development over the past 15 years. Perhaps the single most important landmark has been the approval by the Food and Drug Administration of intravenous (IV) tissue plasminogen activator (t PA) for the treatment of ischemic stroke. However, the approval of this drug has not met with unanimous support by the medical community and, at present, only a minority of stroke patients receive t-PA. Although this is partly due to the fact that many patients do not meet criteria for treatment with IV t-PA, others simply do not arrive at medical facilities sufficiently early to be safely managed using thrombolysis. The appropriate use of IV t-PA in the treatment of ischemic stroke requires proper selection of patients and strict adherence to clinical protocols of treatment. The ideal stroke patient for treatment with IV t-PA is one who suffers occlusion of a small artery that leads to a disabling deficit. PMID- 11389798 TI - Prevention of strokes. AB - Stroke is a major cause of morbidity and mortality. Risk factors for stroke have been determined through prospective epidemiologic study. Control of risk factors has been demonstrated to reduce stroke incidence, either through controlled trials or inferred from observational studies. In the past few years, new approaches to the treatment of established risk factors have been discovered. These include aggressive control of hypertension in diabetes patients, prevention of type 2 diabetes through lifestyle modification, carotid endarterectomy for moderate symptomatic carotid stenosis, encouragement of a high level of physical activity, and control of abdominal obesity and elevated body mass index. In addition, new strategies for stroke prevention have been identified, including encouragement of a diet high in fruits, vegetables, whole grains, and omega-3 fatty acids, the use of vitamins B12, B6, and folic acid in hyperhomocysteinemia, and moderate alcohol consumption. Clinical trial data support the use of hydroxy methyl-coenzyme A inhibitors in patients with coronary artery disease, and ramipril in high-risk patients with coronary artery disease and diabetes, for the primary prevention of stroke. New risk factors for stroke are being investigated, including the role of chronic inflammation and infection, and these may provide future strategies for stroke prevention. PMID- 11389799 TI - Antiphospholipid syndrome, antiphospholipid antibodies, and atherosclerosis. AB - The antiphospholipid syndrome is characterized by arterial and venous thrombosis, as well as pregnancy morbidity, in the presence of elevated levels of antiphospholipid antibodies. These autoantibodies have procoagulant activity, as they affect platelets, humoral coagulation factors, and endothelial cells. In addition, they are proatherogenic, as demonstrated by animal models and by the increased prevalence of cardiovascular diseases in patients with systemic lupus erythematosus and antiphospholipid syndrome. Moreover, antiphospholipid antibodies, including anticardiolipin, anti-b2-glycoprotein-I, and anti-oxidized low-density lipoprotein, are associated with atherosclerosis and its consequences in the general population as well. This autoimmune aspect of atherosclerosis in the presence or absence of an autoimmune disease suggests benefit from development of immunomodulating therapies. PMID- 11389800 TI - Stroke and cognition. AB - Several studies confirm cognitive impairment and dementia to be increased after stroke in the elderly. Although not necessarily involving memory deficits, the frequency of cognitive impairments may occur in up to 30% of stroke survivors at 3 months. This impairment may be confounded by preexisting cognitive decline or dementia. By contrast, cognitive changes and dementia are widely recognized in familial forms of stroke, such as cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL). Several factors, including type of stroke, recurrent episodes, the site and laterality of the lesion(s), volume of cerebral infarction, medial temporal lobe atrophy, and coexistent neurodegenerative pathology predict the degree of impairment. Aphasia, diabetes mellitus, atrial fibrillation, and depression are listed among other biologic factors that further exacerbate cognition and affect long-term survival. There is no clear consensus whether genetic factors, such as the apolipoprotein E e4 allele or angiotensin converting enzyme gene polymorphisms, modify cognitive changes or stroke outcome. Although several neurotransmitter systems may be affected in post-stroke dementia, the amelioration of cholinergic function is a worthy target. PMID- 11389801 TI - Post-stroke epilepsy. AB - Seizures occur in about 10% of stroke patients. Hence, stroke is the most common cause of seizures and epilepsy in the elderly population. Five percent are early onset seizures (peak onset within the first day after the stroke) and another 5% are late-onset seizures (peak onset within 6 to 12 months after the stroke). Epilepsy (i.e., recurrent seizures) develops in 3% to 4% of the stroke patients (in about one third of the patients with early-onset seizures and about one half of the patients with late-onset seizures). There is a strong positive correlation between stroke severity and the risk of post-stroke seizures; the risk is very low in mild strokes. Seizures are more common in hemorrhagic stroke and in stroke with cortical involvement. Whether this is due to the hemorrhagic component or the cortical involvement per se, or a reflection of more severe strokes among patients with hemorrhagic strokes and lesions involving cortical structures, is not clear. The influence of seizures on outcome is still a matter of controversy. Although epileptic seizures are considered easy to control, this is not supported by evidence from randomized controlled trials. PMID- 11389802 TI - Somatization Disorder. AB - There are many new developments regarding somatization disorder, which is among the most difficult and cumbersome of the psychiatric disorders encountered in neurology practice. Diagnostic criteria have been revised to facilitate clinical care and research. The differential diagnosis includes neurologic disorders (eg, multiple sclerosis, epilepsy), systemic medical disorders, and other psychiatric disorders (eg, mood and anxiety disorders, conversion disorder, malingering, and factitious disorder). Many patients have one or more of these illnesses comorbid with somatization disorder. Finally, somatization disorder demands creative biopsychosocial treatment planning by the neurologist, psychiatrist, and other health professionals. PMID- 11389803 TI - Psychiatric Manifestations of Epilepsy in Children and Adolescents. AB - There is a high incidence of psychiatric difficulties in children and adolescents with epilepsy. The severity and range of the problems that exist are broad. In pseudoseizures, which may coexist with true epileptic seizures, the primary problem is psychiatric. Optimal management of children and adolescents with epilepsy involves a team approach, or at least a multidisciplinary effort, with close coordination between neurologist, primary health care provider, mental health professional, psychologist, educational specialist, and other providers (eg, physical therapist), depending on the associated features and particular case. Because many children and adolescents with epilepsy ultimately will be taking psychotropic medication and because of their psychiatric difficulties, a knowledge of interactions between anticonvulsants and psychotropic medication is crucial. PMID- 11389804 TI - Anxiety Disorders in Neurologic Illness. AB - Anxiety disorders frequently occur in individuals with neurologic illness. Anxiety may be a symptom of or a reaction to the neurologic disorder, a medication side effect, or a comorbid condition. The most common anxiety disorders seen in neurologic patients are panic disorder, generalized anxiety disorder, social phobia, and obsessive-compulsive disorder (OCD). Very often, these conditions go unrecognized (and therefore untreated) or are attributed to being a normal response to having a neurologic illness. However, if they are not treated, anxiety disorders can significantly increase morbidity and mortality in neurologic patients. Selective serotonin reuptake inhibitors (SSRIs) should be considered the first-line of pharmacologic treatment, because they offer a broad spectrum of efficacy in the anxiety disorders, are generally well tolerated, and are effective in treating comorbid depression. Benzodiazepines, although most effective for acute anxiety, are now considered adjunctive or second-line treatments. Cognitive behavioral therapy and other behavioral treatments are effective in the treatment of anxiety disorders. They should be considered primary treatments for patients who cannot tolerate or prefer not to take medications. PMID- 11389805 TI - Psychosis Due to Neurologic Conditions. AB - Psychosis arises with considerable frequency in a number of neurologic conditions. The treatment of such patients is often challenging, as many of the treatments for psychosis pose some risk of worsening the underlying neurologic condition. Although psychosis may emerge in the context of any neurologic condition that sufficiently disrupts the functioning of or connections between limbic, paralimbic, frontal, subcortical areas mediating complex sensory perception, interpretation, and thought or language organization, secondary psychoses are most often encountered in patients with Alzheimer's disease (Parkinson's disease receives dopaminomimetic therapies) and epilepsy. Psychosis, and particularly delusions and visual hallucinations, may arise in Alzheimer's disease. Based on the available literature, the first-line therapy for this problem is risperidone 0.5 to 3 mg per day. If this treatment proves unsuccessful, low-dose haloperidol or olanzapine should be considered next. If these treatments prove unsuccessful, quetiapine should then be considered. Finally, clozapine may be useful for treatment-refractory psychosis due to Alzheimer's disease, but due caution is warranted given its considerable anticholinergic properties and potential for worsening cognition in these patients. Although disease-emergent psychosis (paranoid delusions and visual hallucinations) may develop in patients with Parkinson's disease, psychosis due to dopaminomimetic therapy is much more common. When such symptoms develop, the accepted first step is to taper anti-parkinsonian medications were possible. Anticholinergic medications, amantadine, selegiline, and dopamine receptor agonists should be reduced or discontinued, provided that the patient can tolerate changes in motor symptoms attendant to such reductions. When these reductions are not feasible or fail to improve treatment-emergent psychosis, low dose quetiapine or clozapine may be useful. The greatest body of evidence supports the effectiveness of these treatments and their relative lack of adverse effects on motor function. When psychosis develops in the context of epilepsy, the generally accepted first step is to maximize anticonvulsant therapy in an effort to reduce the possible contribution of electrophysiologic disturbances in the described areas to psychotic symptoms. When interictal psychosis persists despite such adjustments, initiation with low-dose atypical antipsychotics carries the least risk of lowering seizure threshold and should be considered. PMID- 11389806 TI - Ocular Myasthenia Gravis. AB - Treatment decisions for ocular myasthenia gravis (OMG) should be based on symptomatology. Local, nonpharmacologic treatment of ptosis or diplopia is successful in relatively few patients, and the majority of patients require drug therapy for satisfactory resolution of their symptoms. Response to anticholinesterase agents is variable, but should always be used as the first line agent or adjunctive therapy in the treatment of OMG. It is unknown whether early treatment of OMG with corticosteroids or other immunosuppressive agents prevents or delays the development of generalized myasthenia, although some observations support this speculation. Corticosteroids are usually necessary for adequate improvement of ophthalmoplegia or ptosis. Surgical correction of ptosis or ocular motility deficits is not recommended for most patients with OMG, because of the fluctuating nature of the deficits and the high rate of recurrent deficits following surgery. All patients with OMG should be screened for the presence of thymus tumors, and thymectomy is recommended for all patients with a thymoma. Some patients with OMG who do not harbor a thymus tumor may also benefit from thymectomy. PMID- 11389807 TI - Dural Fistulas. AB - The treatment of dural fistulas is varied and complex. Treatment decisions require consideration of the nature of the symptoms, the location of the lesion, the complexity of the angioarchitecture, and the risk of progression. Standard treatment modalities include compression therapy, endovascular embolization, neurosurgery, and radiosurgery. Complex lesions will often require multiple different interventions. Because dural fistulas may present with a variety of neurologic and ophthalmologic problems, a team of neurologists, neuro ophthalmologists, neuroradiologists, and neurosurgeons should develop a comprehensive treatment plan. PMID- 11389808 TI - Double Vision. AB - When evaluating a patient with a complaint of double vision, it is important to distinguish monocular versus binocular diplopia, which are differentiated by asking the patient to cover each eye separately. In the setting of binocular double vision, one of the two images disappears when either eye is covered, because diplopia is the result of ocular misalignment. On the other hand, monocular double vision resolves when the affected eye is covered, but remains when the opposite eye is occluded. Causes of monocular diplopia include cataract, refractive error, and retinal disease, which can be managed accordingly by an ophthalmologist. However, an unusual form of monocular double vision can occur in the setting of cortical dysfunction. Cerebral polyopia describes the perception of multiple images and arises from an occipital disturbance. It can occur with migraine headaches and can be accompanied by a homonymous hemianopia. Palinopsia refers to the persistence of an image that is no longer in view (visual perseveration or stroboscopic effect) and results from an occipital lesion as well. The exact mechanism of polyopia and palinopsia are uncertain and both conditions are extremely rare. The majority of this discussion will focus on binocular double vision and its management. The main treatment objective of binocular diplopia is to restore the largest area of single binocular vision. Ideally, patients would be able to achieve single vision in all fields of gaze, but this is not always possible. The majority of patients are treated with either prism lenses or eye muscle surgery. PMID- 11389809 TI - Optic Neuritis. AB - Patients with signs and symptoms consistent with acute monosymptomatic optic neuritis should undergo evaluation with gadolinium-enhanced MRI of the brain and orbits to determine whether or not they are at high risk for the development of clinically definite multiple sclerosis (CDMS). The presence of two or more white matter lesions (3 mm or larger in diameter, at least one lesion periventricular or ovoid) suggests high risk for CDMS, and should prompt immediate treatment as follows: Intravenous methylprednisolone sodium succinate (1 g intravenously per day for 3 days) followed by oral prednisone (1 mg/kg per day for 11 days) with a 4-day taper (20 mg on day 1, 10 mg on days 2 and 4). Interferon beta 1-a, which has been demonstrated to significantly reduce the 3-year probability of the development of CDMS and the development of clinically silent MRI lesions in high risk patients with acute optic neuritis, should be considered following IV methylprednisolone treatment (30 &mgr;g intramuscularly weekly). In monosymptomatic patients with fewer than two white matter lesions by MRI, and in patients for whom a diagnosis of CDMS has been established, treatment with IV methylprednisolone followed by oral prednisone (as outlined), should be considered on an individual basis and may hasten visual recovery, but has not been demonstrated to affect long-term visual outcome. In all cases of typical acute monosymptomatic demyelinating optic neuritis, oral prednisone alone at a dose of 1 mg/kg per day, without prior treatment with IV methylprednisolone (1 g per day for 3 days), may increase the risk for recurrent optic neuritis, and should be avoided. PMID- 11389810 TI - Transitional cell carcinoma of the bladder: new approaches to the treatment of advanced disease. PMID- 11389812 TI - Prevention of non-melanoma skin cancer. AB - Basal cell and squamous cell carcinomas comprise the majority of non-melanoma skin cancers. Whereas the incidence of skin cancer is equivalent to that of all other cancers combined, non-melanoma skin cancer receives a disproportionate share of attention because mortality is relatively low. However, the impact on public health is striking. This review is intended to update readers on the current findings in research on the prevention of these diseases. Topics covered include preventive strategies targeting high-risk populations, chemoprevention (including treatment of intraepithelial neoplasia), and an overview of recent and ongoing clinical and preclinical studies involving new chemopreventive agents. PMID- 11389813 TI - Retinoid mechanisms and cyclins. AB - Retinoids have been investigated for their effects in the prevention and treatment of cancer. Scores of synthetic and natural ligands suppress growth and normalize differentiation of cells in vitro and in vivo. The molecular mechanisms of these activities are being elucidated with the goal of improving the therapeutic index. Here we summarize recent advances in the understanding of retinoid signaling via nuclear receptors, corepressors, and coactivators and review the effects of retinoid treatment on cell-cycle control elements and cyclin proteins. PMID- 11389814 TI - Do antioxidants still have a role in the prevention of human cancer? AB - Antioxidants are components of diet that are involved in DNA and cell maintenance and repair. Dietary antioxidants include carotenoids, vitamin C, vitamin E, and selenium. Across a variety of cancers, the observational studies have inconsistent results with respect to the relationship shown of specific dietary intake or serum levels of antioxidants and risk of certain cancers. The results of the micronutrient supplement trials clearly do not support a reductionist approach to promoting regression of precancerous lesions or prevention of new cancer, except in a few cancers and specific populations. The ability of the antioxidant micronutrients to influence the risk for tissue injury and for cancer, mediated by their antioxidant activities, remains hypothetical. PMID- 11389815 TI - Mammographic densities as a marker of human breast cancer risk and their use in chemoprevention. AB - Differences in the parenchymal pattern of the breast on mammography reflect differences in the amounts of stromal, epithelial, and fat tissue present in the breast. Stroma and epithelium are radiologically dense, whereas fat is lucent. Extensive areas of mammographically dense breast tissue are strongly associated with an increased risk of breast cancer. A variety of interventions, including gonadotropin-releasing hormone inhibitor, tamoxifen, stopping hormone replacement therapy (HRT), and adopting a low-fat, high-carbohydrate diet, all influence the tissue composition of the breast and reduce mammographic densities. Of the interventions examined to date, only tamoxifen has been shown to reduce the incidence of breast cancer, at least in the short term. Conversely, HRT, which increases density, also increases risk of breast cancer. These results suggest that mammographic density may be a short-term marker of the effect on the breast of potential preventive interventions for breast cancer. PMID- 11389816 TI - Prevention of cervical cancer with vaccines. AB - Worldwide, cervical cancer is one of the most common cancers in women. This is especially true in developing countries, where Papanicolaou smear screening, an effective preventive measure against cervical cancer, is insufficiently implemented. With growing evidence for human papillomavirus as a central etiologic factor in cervical neoplasia, development of a vaccine against this virus has emerged as an important objective in prevention of cervical cancer. International efforts in vaccine development have culminated in advancement of various vaccine strategies and initiation of human clinical trials. Reports from animal vaccine trials and early phase I human trials indicate markedly enhanced immune response through vaccination. However, the clinical significance of these results requires confirmation from long-term human trials. PMID- 11389818 TI - Review of the 2001 AJCC staging system for cutaneous malignant melanoma. AB - The American Joint Committee on Cancer (AJCC) staging system for melanoma has recently been revised and published. The previous staging system had not been substantially modified since the late 1980s. In a series of papers, the staging system for melanoma was critically analyzed, and many shortcomings were identified. Many well-established prognostic factors were not used in the staging system. This assessment has led to a substantially modified staging system for cutaneous melanoma in 2001 that is a considerable improvement over past staging systems, albeit more complex. The following modifications are the most important: 1) The primary determinant of tumor (T) staging is tumor thickness as measured in millimeters. The Clark level of invasion is now used only for defining T1 (< or = 1mm) melanomas; 2) The cutpoints for tumor thickness are less than or equal to 1 mm, 1 to 2 mm, 2 to 4 mm, and greater than 4 mm; 3) Ulceration has been added in describing the primary tumor; 4) Local recurrence, satellite disease, and in transit metastases have similar prognosis and are now all classified together as regional stage III disease; 5) Size of lymph node as prognostic factor has been eliminated and replaced with the number of positive nodes; 6) The presence of an elevated serum lactic dehyrogenase level is used in the metastasis (M) category. This revised staging system more precisely defines prognosis and will improve the stratification of patients in future clinical trials. PMID- 11389819 TI - Autologous and allogeneic high-dose therapy for melanoma. AB - Opportunities for the treatment of melanoma with high-dose chemotherapy have been inadequately explored because of the failure of early studies to demonstrate an advantage for high doses of those agents with disease activity that could also be safely dose-escalated without excessive extramedullary toxicities. However, emerging concepts of tumor and transplantation immunology have recently coincided, providing the rationale for new strategies that exploit principles of allogeneic transplant to overcome immunologic tolerance and escape mechanisms in the tumor-bearing individual. It is hoped that this setting will provide an improved milieu for donor-derived immunotherapeutic intervention that takes advantage of shared tumor antigens with therapeutic potential shown in a variety of tumor vaccination studies. PMID- 11389820 TI - Targeted therapy for malignant melanoma. AB - There has been much development in the field of targeted therapy for melanoma stemming from efforts to decrease treatment-related toxicities and enhance specific cytotoxicity. This review focuses on three modalities of targeted melanoma therapy based on the biology of the targeting mechanism. The first of these modalities is immunotherapy, which functions to generate a specific antimelanoma immunity. A second modality utilizes metabolic pathways of melanin synthesis to target melanoma cells specifically. A third modality ensues from recent advances in molecular biology and the identification of genes responsible for the malignant transformation of normal melanocytes to melanomas. This work has furthered our understanding of the basis of malignancy, as well as the development of novel strategies aimed at targeting aberrant growth in melanoma. PMID- 11389821 TI - Angiogenesis and melanoma. AB - Angiogenesis is a process that is central to tumor growth and survival. This process is stimulated by a variety of intrinsic growth factors such as vascular endothelial growth factor, basic and acid fibroblast growth factor, and platelet derived endothelial growth factor, among others. The process of neo-angiogenesis has been shown to be key in the proliferation of melanoma, and primarily believed to be so in the metastatic process. Biologic markers of angiogenesis are being evaluated for correlations with prognosis and biologic behavior of the tumor. These markers may also indicate susceptibility to targeted therapy. Interruption of the tumor-sustaining process of angiogenesis has become a major focus of anticancer drug development. Promising agents are in both preclinical and clinical development. Several may prove to be clinically important. PMID- 11389822 TI - Isolated limb perfusion for extremity soft-tissue sarcomas, in-transit metastases, and other unresectable tumors: credits, debits, and future perspectives. AB - Isolated limb perfusion (ILP) with melphalan is effective against melanoma in transit metastases but has failed in the treatment of limb-threatening extremity sarcomas. Tumor necrosis factor-alpha (TNF) has changed this situation completely. Now, ILP with TNF + melphalan is a very successful treatment to prevent amputation. In a multicenter European trial, ILP with TNF + melphalan resulted in a 76% response rate and a 71% limb salvage rate in patients with limb threatening soft-tissue sarcomas, deemed unresectable by independent review committees, leading to approval of TNF in Europe. We have also reported on the success of this regimen against bulky melanomas, multifocal skin cancers, and drug-resistant bony sarcomas. High-dose TNF destructs tumor vasculature, and, most importantly, it enhances tumor-selective drug uptake (ie, melphalan and doxorubicin) by threefold to sixfold. Similar synergy is observed in well vascularized liver metastases after isolated hepatic perfusion with TNF and melphalan. New (vasoactive) drugs and mechanisms of action and interaction with chemotherapy are in development. ILP is also a promising treatment modality for adenoviral vector-mediated gene therapy. Many clinical phase I/II evaluations in ILP are now underway. PMID- 11389824 TI - Shaping morphogen gradients. PMID- 11389823 TI - Transcription factors and other dysregulated proteins in melanoma prognosis. AB - Approximately one third of patients with cutaneous melanoma later develop a metastatic disease, having then an extremely poor rate of survival. Because of the highly unpredictable nature of melanomas, finding those patients who are likely to develop a metastatic disease and those patients who probably will survive is an ongoing challenge. The current "conventional" prognosticators, such as Breslow thickness, Clark level of invasion, and ulceration, cannot perfectly predict the clinical course of this disease at an individual level. Although the sentinel lymph node biopsy procedure and reverse transcription polymerase chain reaction techniques have significantly improved the staging of patients with melanoma, new molecular prognostic markers may help in selection of appropriate patients for strenuous adjuvant therapies and for randomized clinical trials. Furthermore, these markers also improve our basic understanding of the biology of cutaneous melanoma, potentially offering new targets for novel treatment strategies. This paper reviews the current literature on transcription factors and other dysregulated proteins involved in melanoma prognosis. PMID- 11389825 TI - More than one way to skin a catenin. PMID- 11389826 TI - RNA polymerase clamps down. PMID- 11389827 TI - The first plant genome. PMID- 11389828 TI - A novel cellular phenotype for familial hypercholesterolemia due to a defect in polarized targeting of LDL receptor. AB - Basolateral targeting of membrane proteins in polarized epithelial cells typically requires cytoplasmic domain sorting signals. In the familial hypercholesterolemia (FH)-Turku LDL receptor allele, a mutation of glycine 823 residue affects the signal required for basolateral targeting in MDCK cells. We show that the mutant receptor is mistargeted to the apical surface in both MDCK and hepatic epithelial cells, resulting in reduced endocytosis of LDL from the basolateral/sinusoidal surface. Consequently, virally encoded mutant receptor fails to mediate cholesterol clearance in LDL receptor-deficient mice, suggesting that a defect in polarized LDL receptor expression in hepatocytes underlies the hypercholesterolemia in patients harboring this allele. This evidence directly links the pathogenesis of a human disease to defects in basolateral targeting signals, providing a genetic confirmation of these signals in maintaining epithelial cell polarity. PMID- 11389829 TI - Charcot-Marie-Tooth disease type 2A caused by mutation in a microtubule motor KIF1Bbeta. AB - The kinesin superfamily motor protein KIF1B has been shown to transport mitochondria. Here, we describe an isoform of KIF1B, KIF1Bbeta, that is distinct from KIF1B in its cargo binding domain. KIF1B knockout mice die at birth from apnea due to nervous system defects. Death of knockout neurons in culture can be rescued by expression of the beta isoform. The KIF1B heterozygotes have a defect in transporting synaptic vesicle precursors and suffer from progressive muscle weakness similar to human neuropathies. Charcot-Marie-Tooth disease type 2A was previously mapped to an interval containing KIF1B. We show that CMT2A patients contain a loss-of-function mutation in the motor domain of the KIF1B gene. This is clear indication that defects in axonal transport due to a mutated motor protein can underlie human peripheral neuropathy. PMID- 11389830 TI - Cholesterol modification of sonic hedgehog is required for long-range signaling activity and effective modulation of signaling by Ptc1. AB - Sonic hedgehog (Shh) signaling from the posterior zone of polarizing activity (ZPA) is the primary determinant of anterior-posterior polarity in the vertebrate limb field. An active signal is produced by an autoprocessing reaction that covalently links cholesterol to the N-terminal signaling moiety (N-Shh(p)), tethering N-Shh(p) to the cell membrane. We have addressed the role played by this lipophilic modification in Shh-mediated patterning of mouse digits. Both the distribution and activity of N-Shh(p) indicate that N-Shh(p) acts directly over a few hundred microns. In contrast, N-Shh, a form that lacks cholesterol, retains similar biological activity to N-Shh(p), but signaling is posteriorly restricted. Thus, cholesterol modification is essential for the normal range of signaling. It also appears to be necessary for appropriate modulation of signaling by the Shh receptor, Ptc1. PMID- 11389831 TI - Regulated endocytic routing modulates wingless signaling in Drosophila embryos. AB - Embryos have evolved various strategies to confine the action of secreted signals. Using an HRP-Wingless fusion protein to track the fate of endocytosed Wingless, we show that degradation by targeting to lysosomes is one such strategy. Wingless protein is specifically degraded at the posterior of each stripe of wingless transcription, even under conditions of overexpression. If lysosomal degradation is compromised genetically or chemically, excess Wingless accumulates and ectopic signaling ensues. In the wild-type, Wingless degradation is slower at the anterior than at the posterior. This follows in part from the segmental activation of signaling by the epidermal growth factor receptor, which accelerates Wingless degradation at the posterior, thus leading to asymmetrical Wingless signaling along the anterior-posterior axis. PMID- 11389832 TI - Light and brassinosteroid signals are integrated via a dark-induced small G protein in etiolated seedling growth. AB - Plant growth and development are regulated through coordinated interactions between light and phytohormones. Here, we demonstrate that a dark-induced small G protein, pea Pra2, regulates a variant cytochrome P450 that catalyzes C-2 hydroxylation in brassinosteroid biosynthesis. The cytochrome P450 is dark induced and predominantly expressed in the rapidly elongating zone of etiolated pea epicotyls, where Pra2 is also most abundant. Transgenic plants with reduced Pra2 exhibit a dark-specific dwarfism, which is completely rescued by exogenous brassinolide. Overexpression of the cytochrome P450 results in enhanced hypocotyl growth even in the light, which phenocopies the etiolated hypocotyls. We therefore propose that Pra2 and its orthologs are molecular mediators for the cross-talk between light and brassinosteroids in the etiolation process in plants. PMID- 11389833 TI - Immunity to K1 killer toxin: internal TOK1 blockade. AB - K1 killer strains of Saccharomyces cerevisiae harbor RNA viruses that mediate secretion of K1, a protein toxin that kills virus-free cells. Recently, external K1 toxin was shown to directly activate TOK1 channels in the plasma membranes of sensitive yeast cells, leading to excess potassium flux and cell death. Here, a mechanism by which killer cells resist their own toxin is shown: internal toxin inhibits TOK1 channels and suppresses activation by external toxin. PMID- 11389834 TI - Emi1 is a mitotic regulator that interacts with Cdc20 and inhibits the anaphase promoting complex. AB - We have discovered an early mitotic inhibitor, Emi1, which regulates mitosis by inhibiting the anaphase promoting complex/cyclosome (APC). Emi1 is a conserved F box protein containing a zinc binding region essential for APC inhibition. Emi1 accumulates before mitosis and is ubiquitylated and destroyed in mitosis, independent of the APC. Emi1 immunodepletion from cycling Xenopus extracts strongly delays cyclin B accumulation and mitotic entry, whereas nondestructible Emi1 stabilizes APC substrates and causes a mitotic block. Emi1 binds the APC activator Cdc20, and Cdc20 can rescue an Emi1-induced block to cyclin B destruction. Our results suggest that Emi1 regulates progression through early mitosis by preventing premature APC activation, and may help explain the well known delay between cyclin B/Cdc2 activation and cyclin B destruction. PMID- 11389835 TI - Structure of the replicating complex of a pol alpha family DNA polymerase. AB - We describe the 2.6 A resolution crystal structure of RB69 DNA polymerase with primer-template DNA and dTTP, capturing the step just before primer extension. This ternary complex structure in the human DNA polymerase alpha family shows a 60 degrees rotation of the fingers domain relative to the apo-protein structure, similar to the fingers movement in pol I family polymerases. Minor groove interactions near the primer 3' terminus suggest a common fidelity mechanism for pol I and pol alpha family polymerases. The duplex product DNA orientation differs by 40 degrees between the polymerizing mode and editing mode structures. The role of the thumb in this DNA motion provides a model for editing in the pol alpha family. PMID- 11389836 TI - Multiple transcript cleavage precedes polymerase release in termination by RNA polymerase II. AB - The requirement of poly(A) signals to elicit transcription termination of RNA polymerase II (pol II) is firmly established. However, little else is known about the actual process of pol II transcription termination. Evidence presented in this paper, based on analysis of nascent transcripts of the human beta- and epsilon-globin genes, demonstrates that pol II transcription termination involves two distinct and temporally separate events. The first event, termed pretermination cleavage (PTC), is mediated by sequence tracts located downstream of the poly(A) site which appear to promote heterogeneous cleavage of the nascent transcript. The second event, in which pol II disengages from the DNA template, requires that polymerase has transcribed both a PTC sequence tract and a functional poly(A) site. PMID- 11389837 TI - Nonredundant roles of the mPer1 and mPer2 genes in the mammalian circadian clock. AB - Mice carrying a null mutation in the Period 1 (mPer1) gene were generated using embryonic stem cell technology. Homozygous mPer1 mutants display a shorter circadian period with reduced precision and stability. Mice deficient in both mPer1 and mPer2 do not express circadian rhythms. While mPER2 regulates clock gene expression at the transcriptional level, mPER1 is dispensable for the rhythmic RNA expression of mPer1 and mPer2 and may instead regulate mPER2 at a posttranscriptional level. Studies of clock-controlled genes (CCGs) reveal a complex pattern of regulation by mPER1 and mPER2, suggesting independent controls by the two proteins over some output pathways. Genes encoding key enzymes in heme biosynthesis are under circadian control and are regulated by mPER1 and mPER2. Together, our studies show that mPER1 and mPER2 have distinct and complementary roles in the mouse clock mechanism. PMID- 11389839 TI - Siah-1, SIP, and Ebi collaborate in a novel pathway for beta-catenin degradation linked to p53 responses. AB - Destruction of beta-catenin is regulated through phosphorylation-dependent interactions with the F box protein beta-TrCP. A novel pathway for beta-catenin degradation was discovered involving mammalian homologs of Drosophila Sina (Siah), which bind ubiquitin-conjugating enzymes, and Ebi, an F box protein that binds beta-catenin independent of the phosphorylation sites recognized by beta TrCP. A series of protein interactions were identified in which Siah is physically linked to Ebi by association with a novel Sgt1 homolog SIP that binds Skp1, a central component of Skp1-Cullin-F box complexes. Expression of Siah is induced by p53, revealing a way of linking genotoxic injury to destruction of beta-catenin, thus reducing activity of Tcf/LEF transcription factors and contributing to cell cycle arrest. PMID- 11389840 TI - Siah-1 mediates a novel beta-catenin degradation pathway linking p53 to the adenomatous polyposis coli protein. AB - The adenomatous polyposis coli (APC) tumor-suppressor protein, together with Axin and GSK3beta, forms a Wnt-regulated signaling complex that mediates phosphorylation-dependent degradation of beta-catenin by the proteasome. Siah-1, the human homolog of Drosophila seven in absentia, is a p53-inducible mediator of cell cycle arrest, tumor suppression, and apoptosis. We have now found that Siah 1 interacts with the carboxyl terminus of APC and promotes degradation of beta catenin in mammalian cells. The ability of Siah-1 to downregulate beta-catenin signaling was also demonstrated by hypodorsalization of Xenopus embryos. Unexpectedly, degradation of beta-catenin by Siah-1 was independent of GSK3beta mediated phosphorylation and did not require the F box protein beta-TrCP. These results indicate that APC and Siah-1 mediate a novel beta-catenin degradation pathway linking p53 activation to cell cycle control. PMID- 11389841 TI - Control of cell polarity and chemotaxis by Akt/PKB and PI3 kinase through the regulation of PAKa. AB - We demonstrate that PI3 kinase and protein kinase B (PKB or Akt) control cell polarity and chemotaxis, in part, through the regulation of PAKa, which is required for myosin II assembly. We demonstrate that PI3K and PKB mediate PAKa's subcellular localization, PAKa's activation in response to chemoattractant stimulation, and chemoattractant-mediated myosin II assembly. Mutation of the PKB phosphorylation site in PAKa to Ala blocks PAKa's activation and inhibits PAKa redistribution in response to chemoattractant stimulation, whereas an Asp substitution leads to an activated protein. Addition of the PI3K inhibitor LY294002 results in a rapid loss of cell polarity and the axial distribution of actin, myosin, and PAKa. These results provide a mechanism by which PI3K regulates chemotaxis. PMID- 11389842 TI - Nodal signals to Smads through Cripto-dependent and Cripto-independent mechanisms. AB - Nodal ligands are essential for the patterning of chordate embryos. Genetic evidence indicates that EGF-CFC factors are required for Nodal signaling, but the molecular basis for this requirement is unknown. We have investigated the role of Cripto, an EGF-CFC factor, in Nodal signaling. We find that Cripto interacts with the type I receptor ALK4 via the conserved CFC motif in Cripto. Cripto interaction with ALK4 is necessary both for Nodal binding to the ALK4/ActR-IIB receptor complex and for Smad2 activation by Nodal. We also find that Nodal can inhibit BMP signaling by a Cripto-independent mechanism. Inhibition appears to be mediated by heterodimerization between Nodal and BMPs, indicating that antagonism between Nodal and BMPs can occur at the level of dimeric ligand production. PMID- 11389843 TI - Identification of RFC(Ctf18p, Ctf8p, Dcc1p): an alternative RFC complex required for sister chromatid cohesion in S. cerevisiae. AB - We have identified and characterized an alternative RFC complex RFC(Ctf18p, Ctf8p, Dcc1p) that is required for sister chromatid cohesion and faithful chromosome transmission. Ctf18p, Ctf8p, and Dcc1p interact physically in a complex with Rfc2p, Rfc3p, Rfc4p, and Rfc5p but not with Rfc1p or Rad24p. Deletion of CTF18, CTF8, or DCC1 singly or in combination (ctf18Deltactf8Deltadcc1Delta) leads to sensitivity to microtubule depolymerizing drugs and a severe sister chromatid cohesion defect. Furthermore, temperature sensitive mutations in RFC4 result in precocious sister chromatid separation. Our results highlight a novel function of the RFC proteins and support a model in which sister chromatid cohesion is established at the replication fork via a polymerase switching mechanism and a replication-coupled remodeling of chromatin. PMID- 11389844 TI - DNA replication-dependent formation of joint DNA molecules in Physarum polycephalum. AB - Two-dimensional neutral/neutral agarose gel electrophoresis is used extensively to localize replication origins. This method resolves DNA structures containing replication forks. It also detects X-shaped recombination intermediates in meiotic cells, in the form of a typical vertical spike. Intriguingly, such a spike of joint DNA molecules is often detectable in replicating DNA from mitotic cells. Here, we used naturally synchronous DNA samples from Physarum polycephalum to demonstrate that postreplicative, DNA replication-dependent X-shaped DNA molecules are formed between sister chromatids. These molecules have physical properties reminiscent of Holliday junctions. Our results demonstrate frequent interactions between sister chromatids during a normal cell cycle and suggest a novel phase during DNA replication consisting of transient, joint DNA molecules formed on newly replicated DNA. PMID- 11389845 TI - The 19S regulatory particle of the proteasome is required for efficient transcription elongation by RNA polymerase II. AB - It is generally thought that the primary or even sole activity of the 19S regulatory particle of the 26S proteasome is to facilitate the degradation of polyubiquitinated proteins by the 20S-core subunit. However, we present evidence that the 19S complex is required for efficient elongation of RNA polymerase II (RNAP II) in vitro and in vivo. First, yeast strains carrying alleles of SUG1 and SUG2, encoding 19S components, exhibit phenotypes indicative of elongation defects. Second, in vitro transcription is inhibited by antibodies raised against Sug1, or by heat-inactivating temperature-sensitive Sug1 mutants with restoration of elongation by addition of immunopurified 19S complex. Finally, Cdc68, a known elongation factor, coimmunoprecipitates with the 19S complex, indicating a physical interaction. Inhibition of the 20S proteolytic core of the proteasome has no effect on elongation. This work defines a nonproteolytic role for the 19S complex in RNAP II transcription. PMID- 11389846 TI - Modification of the properties of elongating RNA polymerase by persistent association with nascent antiterminator RNA. AB - Nascent RNA encoded by putL, a cis-acting antitermination site of bacteriophage HK022, increases readthrough of terminators by directly modifying the transcript elongation complex. To characterize the interaction between the antiterminator RNA and RNA polymerase, we stalled the elongation complex downstream of putL and determined the sensitivity of the transcript to ribonuclease cleavage. Part of PutL RNA was protected from cleavage by wild-type polymerase, but not by a mutant with a defect in put-dependent antitermination. We also exposed the stalled complex to oligonucleotides complementary to putL RNA, restarted transcription, and measured antitermination. Some, but not all, complementary oligonucleotides inhibited antitermination. Finally, cleavage of the RNA between putL and the 3' end released putL RNA from the stalled complex and prevented antitermination. PMID- 11389847 TI - Transcriptional termination factors for RNA polymerase II in yeast. AB - The molecular connections between mRNA 3' end processing and transcriptional termination have been investigated in S. pombe using a genetic screen. By this approach, we have identified a RNAP II termination domain in the well-defined cleavage polyadenylation factor called CstF-64 in metazoans and Rna15p in S. cerevisiae. Furthermore, this C-terminal domain interacts with Res2, previously identified as a component of the G1/S transition-specific transcription factor MBF. Deletion of res2 in both fission and budding yeast results in a defect in 3' end formation. This raises the possibility that RNAP II transcriptional termination may in some situations be integrated with cell cycle events. PMID- 11389848 TI - Evolutionarily conserved interaction between CstF-64 and PC4 links transcription, polyadenylation, and termination. AB - Tight connections exist between transcription and subsequent processing of mRNA precursors, and interactions between the transcription and polyadenylation machineries seem especially extensive. Using a yeast two-hybrid screen to identify factors that interact with the polyadenylation factor CstF-64, we uncovered an interaction with the transcriptional coactivator PC4. Both human proteins have yeast homologs, Rna15p and Sub1p, respectively, and we show that these two proteins also interact. Given evidence that certain polyadenylation factors, including Rna15p, are necessary for termination in yeast, we show that deletion or overexpression of SUB1 suppresses or enhances, respectively, both growth and termination defects detected in an rna15 mutant strain. Our findings provide an additional, unexpected connection between transcription and polyadenylation and suggest that PC4/Sub1p, via its interaction with CstF 64/Rna15p, possesses an evolutionarily conserved antitermination activity. PMID- 11389849 TI - HIV-1 infection requires a functional integrase NLS. AB - HIV-1 is able to infect nondividing cells productively in part because the postentry viral nucleoprotein complexes are actively imported into the nucleus. In this manuscript, we identify a novel nuclear localization signal (NLS) in the viral integrase (IN) protein that is essential for virus replication in both dividing and nondividing cells. The IN NLS stimulates the efficient nuclear accumulation of viral DNA as well as virion-derived IN protein during the initial stages of infection but is dispensable for catalytic function. Because this NLS is required for infection irrespective of target cell proliferation, we suggest that interactions between uncoated viral nucleoprotein complexes and the host cell nuclear import machinery are critical for HIV-1 infection of all cells. PMID- 11389850 TI - Localization of the ribosomal protection protein Tet(O) on the ribosome and the mechanism of tetracycline resistance. AB - Tet(O) belongs to a class of ribosomal protection proteins that mediate tetracycline resistance. It is a G protein that shows significant sequence similarity to elongation factor EF-G. Here we present a cryo-electron microscopic reconstruction, at 16 A resolution, of its complex with the E. coli 70S ribosome. Tet(O) was bound in the presence of a noncleavable GTP analog to programmed ribosomal complexes carrying fMet-tRNA in the P site. Tet(O) is directly visible as a mass close to the A-site region, similar in shape and binding position to EF G. However, there are important differences. One of them is the different location of the tip of domain IV, which in the Tet(O) case, does not overlap with the ribosomal A site but is directly adjacent to the primary tetracycline binding site. Our findings give insights into the mechanism of tetracycline resistance. PMID- 11389851 TI - Crystal structure of the atypical protein kinase domain of a TRP channel with phosphotransferase activity. AB - Transient receptor potential (TRP) channels modulate calcium levels in eukaryotic cells in response to external signals. A novel transient receptor potential channel has the ability to phosphorylate itself and other proteins on serine and threonine residues. The catalytic domain of this channel kinase has no detectable sequence similarity to classical eukaryotic protein kinases and is essential for channel function. The structure of the kinase domain, reported here, reveals unexpected similarity to eukaryotic protein kinases in the catalytic core as well as to metabolic enzymes with ATP-grasp domains. The inclusion of the channel kinase catalytic domain within the eukaryotic protein kinase superfamily indicates a significantly wider distribution for this group of signaling proteins than suggested previously by sequence comparisons alone. PMID- 11389852 TI - Transcriptome analysis reveals a role of interferon-gamma in human neointima formation. AB - The most effective immediate cure for coronary stenosis is stent-supported angioplasty. Restenosis due to neointima proliferation represents a major limitation. We investigated the expression of 2435 genes in atherectomy specimens and blood cells of patients with restenosis, normal coronary artery specimens, and cultured human smooth muscle cells (SMCs). Of the 223 differentially expressed genes, 37 genes indicated activation of interferon-gamma (IFN-gamma) signaling in neointimal SMCs. In cultured SMCs, IFN-gamma inhibited apoptosis. Genetic disruption of IFN-gamma signaling in a mouse model of restenosis significantly reduced the vascular proliferative response. Our data suggest an important role of IFN-gamma in the control of neointima proliferation. PMID- 11389853 TI - Pin1 acts catalytically to promote a conformational change in Cdc25. AB - Pin1 is an essential protein that can peptidyl-prolyl-isomerize small phosphopeptides. It has been suggested that Pin1 regulates entry into mitosis by catalyzing the cis/trans-isomerization of prolines on critical protein substrates in response to phosphorylation. We show that Pin1 catalytically generates a conformational change on the mitotic phosphatase Cdc25, as assayed by limited protease digestion, differential reactivity to a phosphoserine-proline-directed monoclonal antibody (MPM-2), and by changes in Cdc25 enzymatic activity. Pin1 catalytically modifies the conformation of Cdc25 at stoichiometries less than 0.0005, and mutants of Pin1 in the prolyl isomerase domain are not active. We suggest that, although difficult to detect, phosphorylation-dependent conformational changes mediated by prolyl isomerization may play an important regulatory role in the cell cycle. PMID- 11389854 TI - A role for AMP-activated protein kinase in contraction- and hypoxia-regulated glucose transport in skeletal muscle. AB - Eukaryotic cells possess systems for sensing nutritional stress and inducing compensatory mechanisms that minimize the consumption of ATP while utilizing alternative energy sources. Such stress can also be imposed by increased energy needs, such as in skeletal muscle of exercising animals. In these studies, we consider the role of the metabolic sensor, AMP-activated protein kinase (AMPK), in the regulation of glucose transport in skeletal muscle. Expression in mouse muscle of a dominant inhibitory mutant of AMPK completely blocked the ability of hypoxia or AICAR to activate hexose uptake, while only partially reducing contraction-stimulated hexose uptake. These data indicate that AMPK transmits a portion of the signal by which muscle contraction increases glucose uptake, but other AMPK-independent pathways also contribute to the response. PMID- 11389855 TI - Genetic and molecular characterization of Skb15, a highly conserved inhibitor of the fission yeast PAK, Shk1. AB - The p21-activated kinase, Shk1, is essential for viability, establishment and maintenance of cell polarity, and proper mating response in the fission yeast, Schizosaccharomyces pombe. Here we describe the characterization of a highly conserved, WD repeat protein, Skb15, which negatively regulates Shk1 in fission yeast. A null mutation in the skb15 gene is lethal and results in deregulation of actin polymerization and localization, microtubule biogenesis, and the cytokinetic machinery, as well as a substantial uncoupling of these processes from the cell cycle. Loss of Skb15 function is suppressed by partial loss of Shk1, demonstrating that negative regulation of Shk1 by Skb15 is required for proper execution of cytoskeletal remodeling and cytokinetic functions. A mouse homolog of Skb15 can substitute for its counterpart in fission yeast, demonstrating that Skb15 protein function has been substantially conserved through evolution. PMID- 11389856 TI - Base-pairing between untranslated regions facilitates translation of uncapped, nonpolyadenylated viral RNA. AB - Translationally competent mRNAs form a closed loop via interaction of initiation factors with the 5' cap and poly(A) tail. However, many viral mRNAs lack a cap and/or a poly(A) tail. We show that an uncapped, nonpolyadenylated plant viral mRNA forms a closed loop by direct base-pairing (kissing) of a stem loop in the 3' untranslated region (UTR) with a stem loop in the 5' UTR. This allows a sequence in the 3' UTR to confer translation initiation at the 5'-proximal AUG. This base-pairing is also required for replication. Unlike other cap-independent translation mechanisms, the ribosome enters at the 5' end of the mRNA. This remarkably long-distance base-pairing reveals a novel mechanism of cap independent translation and means by which mRNA UTRs can communicate. PMID- 11389857 TI - SMN, the product of the spinal muscular atrophy gene, binds preferentially to dimethylarginine-containing protein targets. AB - The survival of motor neurons protein (SMN), the product of the neurodegenerative disease spinal muscular atrophy (SMA) gene, functions as an assembly factor for snRNPs and likely other RNPs. SMN binds the arginine- and glycine-rich (RG) domains of the snRNP proteins SmD1 and SmD3. Specific arginines in these domains are modified to dimethylarginines, a common modification of unknown function. We show that SMN binds preferentially to the dimethylarginine-modified RG domains of SmD1 and SmD3. The binding of other SMN-interacting proteins is also strongly enhanced by methylation. Thus, methylation of arginines is a novel mechanism to promote specific protein-protein interactions and appears to be key to generating high-affinity SMN substrates. It is reasonable to expect that protein hypomethylation may contribute to the severity of SMA. PMID- 11389859 TI - Isolation and characterization of fibroblasts derived from regenerating human periodontal defects. AB - In an attempt to understand better the cells responsible for periodontal regeneration, cells from human gingiva, periodontal ligament and regenerating periodontal defects treated with expanded polytetrafluorethylene membranes were isolated, cultured and characterized. Guided tissue regeneration procedures were carried out on three human volunteers around molar teeth destined for extraction. After a 6-week 'healing phase', fibroblast cell cultures were established from explants of the regenerating soft connective tissue (RTF), as well as from the associated periodontal ligament (PLF) and gingiva (GF). Following stimulation with platelet-derived growth factor-beta (PDGF) and insulin-like growth factor-1 (IGF-1), [3H]thymidine-uptake and dye-binding assays were used to assess the rate of DNA synthesis and cell proliferation, respectively. Northern blotting was used to measure the expression of mRNA for the extracellular matrix proteoglycans decorin, biglycan and versican. The results show that the GF and RTF proliferated more quickly than the PLF. PDGF and IGF-1 were mitogenic for all three cell types. Decorin mRNA expression was stronger in the GF than the RTF and PLF, whereas versican mRNA expression was stronger in the GF and PLF than the RTF. Biglycan mRNA expression was strong in the PLF, moderate in the GF and weak in the RTF. The growth factors did not affect the mRNA expression for biglycan, but they upregulated versican and downregulated decorin mRNA. It can be concluded that RTF exhibits properties characteristic of a reparative phenotype. More specifically, it proliferates faster than PLF, from which it is derived, while exhibiting a unique pattern of proteoglycan mRNA expression. Therefore, this study demonstrates that fibroblasts obtained from the regenerating periodontal defects exhibit characteristics consistent with their ability to facilitate periodontal regeneration. PMID- 11389860 TI - The effect of pilocarpine on salivary constituents in patients with chronic graft versus-host disease. AB - Chronic graft-versus-host disease (cGVHD) is a complex clinical entity with various target organs, including the salivary glands. Oral pilocarpine (Salagen(R)), 30 mg/day, can ameliorate cGVHD-induced xerostomia and improve the flow rate from the major salivary glands. The purpose here was to evaluate the effect of this drug at 30 mg/day on salivary biochemical and immunological composition in cGVHD patients. Significantly higher concentrations of salivary sodium (Na), magnesium (Mg), total protein, albumin, epidermal growth factor (EGF) and total IgG, accompanied by a concomitant increase in total IgA which did not reach significance, were observed in cGVHD patients in comparison with controls, in both resting and stimulated conditions (p < 0.05), while salivary potassium, calcium and phosphate were not altered. Two weeks of oral pilocarpine, at 30 mg/day, resulted in normalization of the altered salivary biochemical and immunological composition in the cGVHD patients. Oral pilocarpine was able to reduce and normalise the elevated Na, Mg, total protein, albumin, EGF, IgG and IgA concentrations in both resting and stimulated conditions. The ability of oral pilocarpine to normalise and reverse the salivary biochemical and immunological alterations induced by cGVHD parallels its known stimulatory effect on salivary flow rates. As the biochemical and immunological composition of saliva provides its protective antimicrobial characteristics, the ability of pilocarpine to abrogate cGVHD salivary gland abnormalities may be of clinical significance. PMID- 11389861 TI - Eroded enamel lesion remineralization by saliva as a possible factor in the site specificity of human dental erosion. AB - The composition and flow of saliva, which determine its functions, vary within intraoral sites and among individuals. Also, the susceptibility to tooth erosion reportedly varies among individuals and within the dental arches. A possible effect of saliva on early-eroded lesions may be a contributory factor. The aims here were firstly to determine the remineralization of eroded enamel lesions by saliva, and secondly to investigate any variation of this remineralization within the dental arches and among individuals. Early enamel erosion was produced on human premolars using orange juice. Control sections and two test slabs were cut from each tooth. The two slabs from the same lesion were bonded with composite resins to the palatal surface of upper right lateral incisor teeth and the lingual surface of the lower right lateral incisor teeth of volunteers, who then chewed a sugar-free gum four times daily. After 28-day intraoral exposure, mineral loss (DeltaZ) and lesion depth (ld) were quantified using microradiography and the data analysed by paired t-test (n=10, alpha=0.05). Mean DeltaZ was significantly lower in the group of slabs positioned palatally (P<0.001) and lingually (P<0.001) when compared with the control group, and in the lingually placed group when compared with the palatally positioned (P<0.01). A significantly lower ld was observed in the group of slabs positioned palatally (P<0.05) and lingually (P<0.001) when compared with the control group, and in the lingually positioned group when compared with the palatally placed (P<0.05). It was concluded that saliva can remineralize early enamel erosion, and that the degree of remineralization varies within intraoral sites and may be responsible for the differing susceptibility to erosion within the dental arches. PMID- 11389862 TI - A randomized clinical trial of the effect of yoghurt on the human salivary microflora. AB - Yoghurt is active against some human pathogens, so this experiment tested whether it is also active against the salivary microflora. A clinical trial was designed, with volunteers aged between 23 and 37 years. Initially, for 8 weeks, they consumed neither yoghurt nor casein-free soybean ice cream (phase 1). They were then split randomly into test (yoghurt) and control (ice cream) groups and required to consume 125 g of these foods twice daily for 8 weeks (phase 2) and then to avoid them for the following 2 weeks (phase 3). Many potential sources of microbial fluctuation were standardized. Salivary samples were then collected at regular intervals of 2 weeks. For each group and phase, the mean logarithms of the salivary counts for total viable flora, oral streptococci, mutans streptococci, lactobacilli and Candida were calculated. The prevalence of Streptococcus thermophilus and Lactobacillus bulgaricus was also assessed. Twenty test and 22 control participants completed the study. At phase 2, the mean for mutans streptococci was lower in the test than in the control group (3.6 vs. 4.0 log colony-forming units/ml; P=0.02). Moreover, the mean had decreased in all test participants with high phase 1 means for lactobacilli as compared to only 36.4% of those with low phase 1 means (P=0.01). L. bulgaricus was transiently detected in three test participants during phase 2. These results suggest that yoghurt does have some activity against the salivary microflora, but this does not seem to be due to the installation of yoghurt microorganisms in the mouth. PMID- 11389863 TI - An immunoelectron-microscopic study of class II major histocompatibility complex molecule-expressing macrophages and dendritic cells in experimental rat periapical lesions. AB - Previous studies have demonstrated that heterogeneous populations of class II major histocompatibility complex (MHC) molecule-expressing non-lymphoid cells, ultrastructurally classified as macrophages and dendritic cell (DC) cell-like cells, comprise the major immune cell population in experimental periapical lesions in rat molars. In this study, the temporal changes in relative proportions of the two types of cells were examined, on the hypothesis that they are involved in different aspects of the pathogenesis of the lesions. The lesions were induced by making surgical pulp exposures in mandibular first molars of 5 week-old Wistar rats. Observation periods were set at 0 (normal), 3, 14, 28, and 56 days. Non-lymphoid cells immunoreactive to OX6 (reactive to class II MHC molecules) were classified as macrophages and DC cell-like cells according to their ultrastructure, and the frequencies of the two types of cells were assessed at each time-point. ED1 (reactive to nearly all macrophages and DCs) was also used to identify macrophages and DC cell-like cells. At 3 days, most OX6+ cells and ED1+ cells in the periapical tissue had the ultrastructural appearance of newly recruited macrophages. At 14 days, when the lesion was actively expanding, there were significantly more OX6+ macrophages than OX6+ DC cell-like cells (P<0.01). However, at 28 days, when lesion expansion had ceased, DC cell-like cells significantly outnumbered OX6+ macrophages (P<0.01); this remained constant at 56 days. Cell-to-cell contact between OX6+ non-lymphoid cells and OX6- lymphocytes, suggesting a functional interaction, was most frequently seen at 28 days. These results support the notion that class II MHC molecule-expressing macrophages play some part in the initial lesion expansion, and suggest that DC cell-like cells may primarily be involved in immune defence against perpetuated antigenic challenges following lesion stabilization. PMID- 11389864 TI - Correlation of the near-infrared spectroscopy signals with signal intensity in T(2)-weighted magnetic resonance imaging of the human masseter muscle. AB - The purpose of this study was to compare and contrast blood volume changes transcutaneously measured using near-infrared (NIR) spectroscopy against water signal intensity changes taken from a transverse T(2)-weighted MR image of the masseter muscle in healthy human subjects before, during and after contraction. Eight healthy non-smoking males with no history of chronic muscle pain or vascular headaches participated (mean age: 23.9+/-0.6 years). The MRI data were gathered using a turbo spin echo sequence (TR: 2300 ms; TE: 90 ms; FOV: 188x300 mm; scanning time: 30 s; slice thickness: 10 mm) and the slice level was set at the mid-point between the origin and insertion of the masseter. Intramuscular haemoglobin (Hb) levels and water content of the right masseter muscle were continuously monitored for 2 min before, 30 s during and 15 min after a maximum voluntary clenching (MVC) task. Both the near-infrared and MRI data were baseline corrected and normalized and mean levels were established and plotted. Plots of the data showed that both near-infrared-based total Hb and T(2)-weighted MRI based signal-intensity levels clearly decreased during contraction and a clear post-contraction rebound response was evident after the contraction. The near infrared data were found to be highly correlated with MRI-based signal-intensity data (Pearson's r=0.909, P<0.0001). In conclusion, these data provide powerful evidence that near-infrared data (total Hb), transcutaneously taken from the masseter muscle in humans, will reflect the intramuscular water signal intensity changes seen using a T(2)-weighted MRI imaging method. PMID- 11389865 TI - Neuropeptide expression in the ferret trigeminal ganglion following ligation of the inferior alveolar nerve. AB - Previous studies have found changes in neuropeptide expression in trigeminal ganglion cells after inferior alveolar nerve (IAN) section. These changes may play a part in the persistent sensory abnormalities that can be experienced after trigeminal nerve injuries. Here, neuropeptide expression after IAN ligation was studied, as this type of injury is thought to be more likely to result in sensory disturbances. The neuropeptides investigated were substance P, calcitonin gene related peptide, enkephalin (ENK), galanin (GAL), neuropeptide Y (NPY) and vasoactive intestinal polypeptide. In anaesthetised adult female ferrets the left IAN was sectioned and the central stump tightly ligated. Recovery was allowed for 3 days, 3 or 12 weeks before perfusion-fixation. In a second procedure, 1 week before perfusion, the IAN was exposed and an injection made central to the injury site, using a mixture of 4% Fluorogold and 4% Isolectin B4 conjugated to horseradish peroxidase, to identify cell bodies with axons in the inferior alveolar nerve and cells with unmyelinated axons within this population, respectively. Control experiments involved tracer injection alone. After harvesting the tissue, sagittal sections were taken from both the right and left ganglia and immunohistochemical staining used to reveal the presence of peptides and Isolectin B4 tracer. The results showed a significant decrease in GAL expression after injury and an increase in ENK and NPY expression. No significant differences were seen in the expression of the other peptides or in the proportion of lectin-positive cells at any time after injury. When compared with previous data, significant differences were found between peptide expression following nerve ligation and nerve section. These results reveal that the changes in neuropeptide expression in the trigeminal ganglion that follow IAN injury are dependent upon the type of injury. The extent to which changes in the central neuropeptide levels contribute to the development of sensory disorders remains to be established. PMID- 11389866 TI - An alternatively spliced Muc10 glycoprotein ligand for putative L-selectin binding during mouse embryonic submandibular gland morphogenesis. AB - Late-gestation (embryonic day 18; E18) mouse submandibular glands (SMG) comprise a network of large and small ducts that terminate in lumen-containing, presumptive acini (terminal buds) expressing unique, cell membrane-associated embryonic mucin. The objective here was to clone and sequence embryonic low molecular-weight SMG mucin, predict its secondary structure, and begin to investigate its possible role in SMG development. Evidence was found that: (1) embryonic low molecular-weight mucin is an alternatively spliced Muc10 gene product, 220 amino acids in size (approximately 25 kDa), rich in potential O glycosylation sites, and variably glycosylated (approximately 40 and 68 kDa); (2) consensus secondary-structure prediction for embryonic low molecular-weight mucin is consistent with a molecule that is anchored to the plasma membrane, directly or indirectly (via a glycolipid), and has a protein core that serves as a scaffold for carbohydrate presentation; (3) embryonic L-selectin is immunolocalized to the plasma membrane region of terminal-bud epithelial cells in a pattern similar to that seen for embryonic mucin; (4) embryonic, but not adult, mucin is able to bind L-selectin and does so endogenously in E18 SMG. As the primary role of L-selectin is to mediate cell adhesion and its ligands are mucin like glycoproteins, it is suggested that this embryonic low molecular-weight mucin be termed MucCAM. PMID- 11389867 TI - Isolation and characterisation of dipeptidyl peptidase IV from Prevotella loescheii ATCC 15930. AB - A proline-specific dipeptidyl aminopeptidase, dipeptidyl peptidase IV (EC 3.4.14.5), was purified from a cell sonicate soluble fraction of Prevotella loescheii ATCC 15930 by sequential column chromatography. The molecular mass of the native enzyme was estimated as 160 kDa by high-pressure liquid gel filtration column chromatography and unheated sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). The subunit molecular mass was 80 kDa when the enzyme was heated to 100 degrees C in the presence of 2-mercaptoethanol before SDS-PAGE, suggesting that the native enzyme consists of two identical subunits and is folded in 2% SDS. The optimum pH, with glycyl-prolyl-4-methyl-coumaryl-7-amide as the substrate, was 8.0; the isoelectric point was 5.2. Purified enzyme showed a strong preference for dipeptide substrates containing proline and, less efficiently, alanine in the P1 position. The enzyme was markedly inhibited by Cd(2+), Zn(2+), Hg(2+), Co(2+), and serine proteinase inhibitor di isopropylfluorophosphate. PMID- 11389868 TI - Regulation of hyaluronan synthase gene expression in human periodontal ligament cells by tumour necrosis factor-alpha, interleukin-1beta and interferon-gamma. AB - Accumulation and fragmentation of hyaluronic (HA) accompanies the inflammatory changes in the periodontium and gingival crevicular fluid are involved in periodontitis, but the mechanism for this is unknown. Recently, three human hyaluronan-synthase (HAS1, 2, and 3) genes have been cloned and characterised as synthesising hyaluronans of different molecular weights. Both HAS1 and HAS2 synthesise high molecular-weight HA, whereas HAS3 produces lower molecular weight HA. In the present study the regulation of HAS genes by cytokines in cultured human periodontal ligament (PDL) cells was investigated using a novel real-time fluorescence polymerase chain reaction detection system. Human PDL cells derived from premolars were cultured with or without tumour necrosing factor (TNF)-alpha (1-100 ng/ml), interleukin (IL)-1beta (0.1-10 ng/ml) and interferon (IFN)-gamma (1-100 ng/ml). Expression of HAS mRNA was assessed in cultured cells treated with these cytokines for 0-24 h. The expression of HAS2 mRNA was enhanced about 4.5- and 2.2-fold at maximum after 3-h stimulation with 10 ng/ml TNF-alpha and 1 ng/ml IL-1beta, respectively, whereas IFN-gamma exerted little effect on HAS2 or HAS3 mRNA expression during the experiment. Expression of HAS3 mRNA was increased by about 14- and 10-fold after 3-h stimulation with 10 ng/ml TNF-alpha and 1 ng/ml IL-1beta, respectively. These results suggest that TNF-alpha and IL-1beta regulate HAS expression, and consequently may result in an accumulation of HA and an increase in HA of a lower molecular-weight. PMID- 11389869 TI - Fatigue and pain in human jaw muscles during a sustained, low-intensity clenching task. AB - Fatigue, pain and changes in the electromyographic (EMG) activity of the jaw closing muscles are well documented during short, high-intensity tooth-clenching tasks but less so during sustained, low-intensity tasks. In this study, 11 healthy men clenched on a bite-force meter for 60 min at 10% of the maximal voluntary contraction (MVC) and scored the intensity of fatigue and pain on separate 10 cm visual analogue scales (VAS). Surface EMG activity from the masseter and anterior temporalis muscles was recorded in 10 s epochs every 5 min throughout the task. Pressure-pain thresholds (PPTs) in the jaw-closing muscles, unassisted maximum jaw opening and MVC were determined before and after the task. All participants reported an increasing sensation of fatigue in the jaw-closing muscles during the task (mean+/-SD: peak VAS=7.5+/-2.0 cm) but all were able to maintain the required force. Most (7/11) also reported a painful sensation (peak VAS=2.7+/-2.8 cm). The jaw-opening capacity (59.5+/-7.4 vs. 58.3+/-6.5 mm, P=0.031) and the MVC (777+/-73 vs. 652+/-115 N,P=0.002) were slightly, but significantly, decreased immediately after the task whereas the PPTs remained unchanged (ANOVA: P=0.612). The mean frequency of the EMG activity decreased in all muscles during the task (95.7 vs. 46.6 Hz;P<0.001), and the root mean squares increased (53.2 vs. 154 microV, P<0.001). The changes in EMG activity were more strongly correlated with the sensation of fatigue than pain. These findings demonstrate that a sustained, low-intensity clenching task can induce subjective and electrophysiological indications of fatigue. PMID- 11389872 TI - Molecular genetics of nucleoside transporters in Leishmania and African trypanosomes. AB - Nucleoside transporters play central roles in the biochemistry of parasitic protozoa such as Leishmania and African trypanosomes, because these parasites cannot synthesize purines de novo and are absolutely reliant upon purine salvage from their hosts. Furthermore, nucleoside transporters are important to the pharmacology of these significant human pathogens, because they mediate the uptake of purine analogs, as well as some non-purine drugs, that are selectively cytotoxic to the parasites. Recent advances in molecular biology and genomics have allowed the cloning and functional expression of several nucleoside transporter genes from Leishmania donovani and Trypanosoma brucei, providing molecular reagents for a detailed functional examination of these permeases and their role in the delivery of nutrients and drugs to the parasites. Furthermore, the molecular basis of drug-resistant mutants that are deficient in nucleoside transport functions can now be fathomed. PMID- 11389873 TI - Major roles of prostanoid receptors IP and EP(3) in endotoxin-induced enhancement of pain perception. AB - To know the roles of prostaglandin I (IP) and prostaglandin E (EP) receptors in pain perception, we compared the acetic acid-induced writhing response in mice deficient in prostaglandin receptors, i.e. IP, EP(1,) EP(2,) EP(3,) or EP(4,) with or without lipopolysaccharide (LPS) pretreatment. Without LPS pretreatment, IP-receptor deficient mice showed a significantly smaller number of responses, as previously reported, whereas mice deficient in any of the EP-receptor subtypes showed a number of writhings similar to those of wild-type mice. When mice were pretreated with LPS for 24 hr to induce cyclooxygenase-2 expression, the wild type as well as EP(1)-, EP(2)-, or EP(4)-receptor-deficient mice showed a similar enhanced writhing response, whereas IP- and EP(3)-receptor-deficient mice had a significantly less enhanced number of writhings. These results indicate that IP and EP(3) are the major prostaglandin receptors mediating the enhanced acetic acid-induced writhing response in mice pre-exposed to LPS, i.e. in endotoxin enhanced inflammatory nociception. PMID- 11389874 TI - Sex-associated expression of mouse hepatic and renal CYP2B enzymes by glucocorticoid hormones. AB - The expression of Cyp2b9 and Cyp2b10 genes was investigated in kidney, liver, and cultured hepatocytes of adult C57BL/6NCrj mice. The constitutive expression level of CYP2B mRNA in kidney was higher in female than in male mice, as it was in the liver where more CYP2B9 than CYP2B10 was expressed in the females, and more CYP2B10 was expressed in the males. After treatment with dexamethasone (Dex), induction of CYP2B10 mRNA and protein in the kidneys was far greater in male than in female mice. In contrast to Dex, phenobarbital (PB), pregnenolone-16 alpha carbonitrile (PCN), and 1,1,1-trichloro-2,2-bis(p-chlorophenyl)ethane (DDT) did not induce the expression of the Cyp2b gene in the kidneys of either sex. In the liver, PB, PCN, and DDT induced both CYP2B9 and CYP2B10 in both sexes to the same extent, whereas Dex induced only CYP2B10 and simultaneously suppressed CYP2B9. Dex-inducible expression of CYP2B mRNA was decreased by 11 beta-[4 dimethylamino]phenyl-17 beta-hydroxy-17-[1-propynyl]estra-4,9-dien-3-one (RU 486), in both the kidneys and liver from male mice, and in cultured hepatocytes. However, RU-486 itself induced the expression of CYP2B mRNA in female liver and cultured hepatocytes. Interestingly, RU-486 increased PB-inducible expression of these species in cultured hepatocytes. Gonadectomy increased the expression of CYP2B mRNA in untreated male liver, but suppressed Dex-induced expression in the kidneys of both sexes. These observations suggest that (a) there are multiple regulatory pathways in the expression of Cyp2b genes, one of which used by Dex is mediated via the glucocorticoid receptor, which is different from that used by PB, and (b) sex hormones play a role in the regulation of the sex-dependent expression of Cyp2b genes in the mouse. PMID- 11389875 TI - Modulation by cellular cholesterol of gene transcription via the cyclic AMP response element. AB - The effect of rapid changes in cellular cholesterol content on adenosine 3',5' cyclic monophosphate (cAMP) response element-mediated gene transcription was investigated. The study was carried out in Chinese hamster ovary (CHO-K1) cells permanently expressing the human beta(2)-adrenoceptor. Gene transcription was quantified using a reporter gene (secreted placental alkaline phosphatase) under the transcriptional control of cAMP response element (CRE) sequences. Cellular cholesterol was reduced by 42% or elevated by 47% by incubating cells for 1 hr with methyl-beta-cyclodextrin alone or methyl-beta-cyclodextrin complexed with cholesterol, respectively. There was a significant negative correlation between the free cholesterol content of the cells and CRE-mediated gene expression in response to 10(-6) M isoprenaline (slope = -4.57 +/- 0.73, P < 0.001), indicating that beta(2)-adrenoceptor-mediated activation of the CRE is inhibited by cholesterol. Cyclic AMP accumulation in response to isoprenaline (10(-12) to 10( 5) M) was also inhibited in cholesterol-enriched cells and enhanced in cholesterol-depleted cells compared to controls (P < 0.05, two-way ANOVA). Cholesterol also inhibited serum-mediated enhancement of CRE-driven gene expression, and we present data suggesting that the pathway activated by serum and inhibited by cholesterol could be independent of adrenoceptor activation and protein kinase A. We conclude that in CHO-K1 cells cholesterol inhibits at least two processes that can stimulate CRE-mediated gene expression. One is isoprenaline activation of cAMP synthesis, the other is activated by serum. These findings demonstrate that activation of gene transcription by extracellular stimuli could be influenced by cellular cholesterol content. PMID- 11389876 TI - Beta gamma-mediated enhancement of corticotropin-releasing hormone-stimulated adenylyl cyclase activity by activation of gamma-aminobutyric acid(B) receptors in membranes of rat frontal cortex. AB - A number of studies have shown that activation of gamma-aminobutyric acid(B) (GABA(B)) receptors potentiates neurotransmitter-induced accumulation of cyclic AMP in brain slices, but the mechanisms involved in the facilitatory effect have not been fully elucidated. In the present study, we showed that in membranes of rat frontal cortex the GABA(B) receptor agonist (-)baclofen increased basal adenylyl cyclase activity and potentiated the maximal enzyme stimulation elicited by corticotropin-releasing hormone (CRH). The less active enantiomer (+)baclofen had no effect on cyclic AMP formation, whereas the natural agonist GABA mimicked the stimulatory action of (-)baclofen. In radioligand-binding experiments, the affinity and maximal binding capacity of (125)I-Tyr-CRH was not affected by ( )baclofen. The GABA(B) receptor antagonist CGP 55845A competitively counteracted the (-)baclofen potentiation of CRH-stimulated adenylyl cyclase activity with a pA(2) value of 6.70. Moreover, both (-)baclofen and GABA, but not (+)baclofen, caused a concentration-dependent stimulation of [(35)S]GTP gamma S binding to membrane G-proteins. The intracerebral injection of pertussis toxin significantly reduced the facilitatory effects of (-)baclofen on both basal and CRH-stimulated adenylyl cyclase activities. Moreover, membrane incubation with the GDP-bound form of the alpha subunit of transducin, a scavenger of G protein beta gamma subunits, blocked the stimulatory effects of (-)baclofen. The data indicate that in rat frontal cortex activation of GABA(B) receptors potentiates the CRH stimulation of adenylyl cyclase activity through a mechanism involving the beta gamma subunits of the pertussis toxin-sensitive G protein G(i)/G(o). PMID- 11389877 TI - Troglitazone quinone formation catalyzed by human and rat CYP3A: an atypical CYP oxidation reaction. AB - Oxidative ring opening of troglitazone (TGZ)(1) a thiazolidine 2,4-dione derivative used for the treatment of type II diabetes mellitus, leads to the formation of a quinone metabolite. The formation of TGZ quinone was shown to be NADPH dependent and to require active microsomal enzymes. Quinone formation was not affected by co-incubation with catalase or sodium azide and was partially inhibited (25%) by superoxide dismutase (SOD). Kinetic analysis of TGZ quinone formation in human liver microsomes implied single enzyme involvement. CYP3A isoforms were characterized as the primary enzymes involved in quinone formation by several lines of evidence including: (a) troleandomycin and ketoconazole almost completely inhibited microsomal quinone formation when SOD was present, whereas other CYP inhibitors had minimal effects (<20%); (b) TGZ quinone formation was highly correlated with regard to both contents (r(2): 0.9374) and activities (r(2): 0.7951) of CYP3A4 in human liver microsomes (HLM); (c) baculovirus insect cell-expressed human CYP3A4 was able to catalyze TGZ quinone formation at a higher capacity (V(max)/K(m)) than other human CYPs with the relative contribution of CYP3A4 in HLM estimated to be 20-fold higher than that of other CYPs; (d) TGZ quinone formation was increased by 350% in liver microsomes from rats pretreated with dexamethasone (DEX); and (e) plasma concentrations of TGZ quinone were increased by 260-680% in rats pretreated with DEX. The chemical nature of the quinone metabolite suggests an atypical CYP reaction consistent with a one-electron oxidation mechanism where an intermediate phenoxy radical combines with ferryl oxygen to subsequently form the quinone metabolite. PMID- 11389878 TI - Influences of glutathione on anionic substrate efflux in tumour cells expressing the multidrug resistance-associated protein, MRP1. AB - The ATP-dependent transport of natural product drugs, e.g. vincristine, by multidrug resistance-associated protein (MRP1) requires reduced glutathione (GSH), whilst that of anionic substrates does not. The present results suggest, however, that GSH can modulate transport of anionic species. Efflux of fluorescent anionic substrates was measured from adherent MRP1-expressing human multidrug-resistant lung tumour cells, COR-L23/R, and drug-sensitive parental cells. As expected, much greater efflux of calcein, methylfluorescein-glutathione (GS-MF), and 2',7'-bis-(2-carboxyethyl)-5-(and-6)-carboxyfluorescein (BCECF) was observed from the resistant cells. Unexpectedly, lowering GSH levels in COR-L23/R cells by inhibiting GSH synthesis with buthionine sulfoximine decreased efflux of calcein and of GS-MF (3-fold and 1.6-fold) but not efflux of BCECF. Transport of the anionic conjugate dinitrophenyl-glutathione ([(3)H]DNP-SG) was investigated by following its uptake into inside-out plasma membrane vesicles prepared from the MRP1-expressing cells. At least 90% of the ATP-dependent uptake was blockable by the anti-MRP1 antibody QCRL-3 and 100 microM vincristine inhibited uptake but only in the presence of 1--3 mM GSH, suggesting MRP1 to be the protein primarily responsible for this transport. Agents shown to reduce efflux of calcein from resistant cells, i.e. indomethacin, MK-571, and probenecid, also inhibited [(3)H]DNP-SG uptakes, consistent with MRP1 being responsible for export of calcein. At concentrations achievable within cells, GSSG (70 microM) inhibited uptake whereas GSH (1 and 3 mM) enhanced uptake. We suggest that variations in both GSH and GSSG levels within cells may affect MRP1-mediated anion transport. PMID- 11389879 TI - Cytochrome P450 CYP1B1 protein expression: a novel mechanism of anticancer drug resistance. AB - The overexpression of human cytochrome P450 CYP1B1 has been observed in a wide variety of malignant tumours, but the protein is undetectable in normal tissues. A number of cytochrome P450 enzymes are known to metabolise a variety of anticancer drugs, and the consequence of cytochrome P450 metabolism is usually detoxification of the drug, although bioactivation occurs in some cases. In this study, a Chinese hamster ovary cell line expressing human cytochrome P450 CYP1B1 was used to evaluate the cytotoxic profile of several anticancer drugs (docetaxel, paclitaxel, cyclophosphamide, doxorubicin, 5-fluorouracil, cisplatin, and carboplatin) commonly used clinically in the treatment of cancer. The MTT (3 [4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium bromide) assay was used to determine the levels of cytotoxicity. The key finding of this study was that on exposure to docetaxel, a significant decrease in sensitivity towards the cytotoxic effects of docetaxel was observed in the cell line expressing CYP1B1 compared to the parental cell line (P = 0.03). Moreover, this difference in cytotoxicity was reversed by co-incubation of the cells with both docetaxel and the cytochrome P450 CYP1 inhibitor alpha-naphthoflavone. This study is the first to indicate that the presence of CYP1B1 in cells decreases their sensitivity to the cytotoxic effects of a specific anticancer drug. PMID- 11389880 TI - Activation pathways of 5-fluorouracil in rat organs and in PC12 cells. AB - Activation of the pyrimidine analogue 5-fluorouracil (5-FU) to the ribonucleotide level may occur through one of the following three pathways: 1) the 5 phosphoribosyl 1-pyrophosphate (PRPP)-mediated direct transfer of ribose 5 phosphate to 5-FU as catalysed by orotate phosphoribosyltransferase; 2) the ribose 1-phosphate (Rib1-P)-mediated addition of ribose by uridine phosphorylase, followed by the action of uridine kinase; and 3) the 2'-deoxyribose 1-phosphate (deoxyRib1-P)-mediated addition of deoxyribose, thought to be catalysed by thymidine phosphorylase, followed by the action of thymidine kinase. Many of the conclusions as to the precise pathways by which normal tissues and different cell lines activate uracil are indirectly derived from drug interactions affecting the availability of the substrates of the three pathways, or from measurement of activities of the enzymes metabolising 5-FU in normal tissues and tumours. In previous papers (Cappiello et al. Biochim Biophys Acta 1998;1425:273--81; Mascia et al. Biochim Biophys Acta 1999;1472:93--8), we assessed the molecular mechanisms by which the natural base uracil is salvaged in vitro to uracil ribonucleotides and deoxyribonucleotides in rat liver and brain. In this paper, we investigated the pathways of 5-FU activation to cytotoxic ribonucleotide and deoxyribonucleotide levels in normal rat tissues and PC12 cell extracts. The results clearly showed that normal rat tissues activated 5-FU mainly via the Rib1 P pathway, and to a lesser extent via the PRPP pathway. The deoxyRib1-P pathway was absent. PC12 cells activated 5-FU mainly via the PRPP pathway and to a lesser extent by the other two pathways. PMID- 11389881 TI - Enzymatic activation of autotaxin by divalent cations without EF-hand loop region involvement. AB - Autotaxin (ATX) is a recently described member of the nucleotide pyrophosphatase/phosphodiesterase (NPP) family of proteins with potent tumor cell motility-stimulating activity. Like other NPPs, ATX is a glycoprotein with peptide sequences homologous to the catalytic site of bovine intestinal alkaline phosphodiesterase (PDE) and the loop region of an EF-hand motif. The PDE active site of ATX has been associated with the motility-stimulating activity of ATX. In this study, we examined the roles of the EF-hand loop region and of divalent cations on the enzymatic activities of ATX. Ca(2+) or Mg(2+) was each demonstrated to increase the PDE activity of ATX in a concentration-dependent manner, whereas incubation of ATX with chelating agents abolished this activity, indicating a requirement for divalent cations. Non-linear regression analysis of enzyme kinetic data indicated that addition of these divalent cations increases reaction velocity predominantly through an effect on V(max.) Three mutant proteins, Ala(740)-, Ala(742)-, and Ala(751)-ATX, in the EF-hand loop region of ATX had enzymatic activity comparable to that of the wild-type protein. A deletion mutation of the entire loop region resulted in slightly reduced PDE activity but normal motility-stimulating activity. However, the PDE activity of this same deletion mutant remained sensitive to augmentation by cations, strongly implying that cations exert their effect by interactions outside of the EF-hand loop region. PMID- 11389882 TI - Cyclic GMP excretion blocked by isatin administration under conditions of fluid overload. AB - Isatin is a potent inhibitor of atrial natriuretic peptide (ANP) receptors and ANP-induced generation of cGMP in vitro. This study was designed to determine whether it had a similar effect in vivo, using a model of fluid overload known to induce ANP. We confirmed that this model increased urinary output of cGMP 3 hr after volume loading, and showed that this effect was blocked by i.p. injection of isatin (50 mg/kg). Isatin had no effect on urine volume or sodium output. However, isatin did have an effect on plasma protein concentration, both compared with control values, compatible with shifting fluid to the vascular compartment, and after volume overload, in which it normalised such a shift. Isatin thus affected both the generation of cGMP and fluid balance in vivo. PMID- 11389883 TI - Prostaglandin D(2) receptor-mediated desensitization of the alpha isoform of the human thromboxane A(2) receptor. AB - Thromboxane (TX) A(2) and prostaglandin (PG) D(2) mediate opposing actions in platelets and in vascular and non-vascular smooth muscle. Here, we investigated the effects of stimulation of the PGD(2) receptor (DP) on signaling by the TXA(2) receptor (TP) expressed in human platelets and in human embryonic kidney (HEK) 293 cells over-expressing the individual TP alpha and TP beta isoforms. In platelets, the selective DP agonist BW245C abolished TP-mediated mobilization of intracellular calcium ([Ca(2+)](i)) and inhibited platelet aggregation in response to the TXA(2) mimetic U46619. DP-mediated desensitization of TP signaling in platelets was prevented by pretreatment with the cAMP-dependent PKA inhibitor, H-89, but was unaffected by the PKC inhibitor GF 109203X. In HEK 293 cells, signaling by TP alpha, but not TP beta, was subject to DP-mediated desensitization in a PKA-dependent, PKC-independent manner. U46619-induced signaling by TP(Delta 328), a truncated variant of TP containing only those residues common to TP alpha and TP beta, was insensitive to prior DP stimulation, indicating that the carboxyl terminal tail of TPalpha contains the target site(s) for DP-mediated desensitization. Mutation of Ser(329) to Ala(329) within a consensus PKA site in TP alpha rendered the mutant TP alpha(S329A) insensitive to BW245C-mediated desensitization. Whole cell phosphorylation assays established that TP alpha, but not TP beta or TP alpha(S329A), was subject to DP-mediated phosphorylation and that TP alpha phosphorylation was blocked by the PKA inhibitor H-89. These data establish that TP alpha, but not TP beta, is subject to DP-mediated cross desensitization, which occurs through direct PKA-mediated phosphorylation of TP alpha at Ser(329). PMID- 11389884 TI - Radical-scavenging and iron-chelating properties of carvedilol, an antihypertensive drug with antioxidative activity. AB - Carvedilol, an antihypertensive agent, has been in clinical use for several years. In addition to its function as a beta-blocker, carvedilol has been shown to act as an antioxidant. However, there is some controversy as to how carvedilol achieves its antioxidative ability: by radical scavenging or ion chelation? We therefore used a method of radical generation independent of metal ions to investigate the antioxidative properties of carvedilol. We showed that carvedilol decreased low-density lipoprotein (LDL) oxidation induced by a peroxyl radical generating system [2,2'-azobis(2-amidinopropane)hydrochloride]. Formation of thiobarbituric acid-reactive substances, lipid hydroperoxides, and newly generated epitopes on oxidised LDL was used to monitor LDL oxidation. We further showed that carvedilol was consumed during reaction with peroxyl radicals. However, carvedilol showed no reaction with nitrogen-centered radicals (1,1 diphenyl-2-picrylhydrazyl and 2,2'-azino-di-[3-ethylbenzthiazoline sulphonate]), which are often used in assays for determining antioxidative properties. On the other hand, we found that carvedilol acted as a chelator of ferric ions. Using mass spectrometry and NMR spectroscopy, we observed complex formation with free and acetylacetonate-complexed ferric ions. The binding constant with Fe(3+) was in the range of 10(5) L/mol. From our data, we concluded that carvedilol acts as both a metal chelator and a radical scavenger in vitro. However, it is selective in reacting with different radicals and is not an electron-donating radical scavenger as is alpha-tocopherol. Therefore, taking into account the low physiological concentration, the antioxidative properties reported earlier may not solely be explained by its radical-scavenging activity. PMID- 11389885 TI - Expression and induction of cytochrome P450s in rabbit parotid glands. AB - Earlier, we isolated and purified five different P450 isoforms from rabbit kidney cortex microsomes, three of which are members of the CYP4A subfamily (CYP4A5, CYP4A6, and CYP4A7), with the others being CYP2B4 and CYP1A1. In contrast, P450s in parotid glands were unknown. The fact that the parotid glands bear a marked morphological and functional resemblance to kidney tissue prompted us to investigate P450s in these glands. The present study was undertaken to determine which P450 isoforms are expressed in this tissue. Microsomes from parotid glands of untreated rabbits were found to contain 42.3 pmol of P450/mg protein and to catalyze the omega-hydroxylation of laurate. Administration of di(2-ethylhexyl) phthalate (DEHP) resulted in a 7-fold increase of laurate omega-hydroxylation. This enzyme activity was greatly inhibited by pretreatment with antibodies against CYP4A5. Furthermore, parotid gland CYP4A5, CYP4A6, and CYP4A7 mRNAs were identified by RT-PCR. Moreover, the CYP4A enzymes were demonstrated immunohistochemically to be localized exclusively in the ducts of these glands. In addition to the CYP4A enzymes, immunoblot analysis revealed that CYP2B4 is constitutively present, and that CYP1A1 is induced in these glands by treatment with 3-methylcholanthrene. Taken together, we can conclude that the P450 isoforms expressed in rabbit kidney cortex and parotid glands are identical in composition. PMID- 11389886 TI - Oxidation of hydrogen sulfide and methanethiol to thiosulfate by rat tissues: a specialized function of the colonic mucosa. AB - Colonic bacteria release large quantities of the highly toxic thiols hydrogen sulfide (H(2)S) and methanethiol (CH(3)SH). These gases rapidly permeate the colonic mucosa, and tissue damage would be expected if the mucosa could not detoxify these compounds rapidly. We previously showed that rat cecal mucosa metabolizes these thiols via conversion to thiosulfate. The purpose of the present study in rats was to determine if this conversion of thiols to thiosulfate is (a) a generalized function of many tissues, or (b) a specialized function of the colonic mucosa. The tissues studied were mucosa from the cecum, right colon, mid-colon, ileum, and stomach; liver; muscle; erythrocytes; and plasma. The metabolic rate was determined by incubating homogenates of the various tissues with H(2)(35)S and CH(3)(35)SH and measuring the rate of incorporation of (35)S into thiosulfate and sulfate. The detoxification activity of H(2)S (expressed as nmol/mg per min) that resulted in thiosulfate production was at least eight times greater for cecal and right colonic mucosa than for the non-colonic tissues. Thiosulfate production from CH(3)SH was at least five times more rapid for cecal and right colonic mucosa than for the non-colonic tissues. We conclude that colonic mucosa possesses a specialized detoxification system that allows this tissue to rapidly metabolize H(2)S and CH(3)SH to thiosulfate. Presumably, this highly developed system protects the colon from what otherwise might be injurious concentrations of H(2)S and CH(3)SH. Defects in this detoxification pathway possibly could play a role in the pathogenesis of various forms of colitis. PMID- 11389887 TI - Differential inheritance modes of DNA methylation between euchromatic and heterochromatic DNA sequences in ageing fetal bovine fibroblasts. AB - To elucidate overall changes in DNA methylation occurring by inappropriate epigenetic control during ageing, we compared fetal bovine fibroblasts and their aged neomycin-resistant versions using bisulfite-PCR technology. Reduction in DNA methylation was observed in euchromatic repeats (18S-rRNA/art2) and promoter regions of single-copy genes (the cytokeratin/beta-lactoglobulin/interleukin-13 genes). Contrastingly, a stable maintenance of DNA methylation was revealed in various heterochromatic sequences (satellite I/II/alphoid and Bov-B). The differential inheritance mode of DNA methylation was confirmed through the analysis of individual neomycin-resistant clones. These global, multi-locus analyses provide evidence on the tendency of differential epigenetic modification between genomic DNA regions during ageing. PMID- 11389888 TI - Masking of the Translin/Trax complex by endogenous RNA. AB - Translin and Trax are components of an RNA binding complex initially detected in extracts of brain and testes. Although other tissues appear to contain much lower or negligible levels of the Translin/Trax gel-shift complex, we found, unexpectedly, that several of these peripheral tissues express Translin and Trax proteins at levels comparable to those present in brain. In this study, we demonstrate that the paradoxically low levels of the Translin/Trax complex in kidney and other peripheral tissues are due to masking of these sites by endogenous RNA. Thus, these findings indicate that the Translin/Trax complex is involved in RNA processing in a broader range of tissues than previously recognized. PMID- 11389889 TI - Expression of fluorescently tagged connexins: a novel approach to rescue function of oligomeric DsRed-tagged proteins. AB - A novel, brilliantly red fluorescent protein, DsRed has become available recently opening up a wide variety of experimental opportunities for double labeling and fluorescence resonance electron transfer experiments in combination with green fluorescent protein (GFP). Unlike in the case of GFP, proteins tagged with DsRed were often found to aggregate within the cell. Here we report a simple method that allows rescuing the function of an oligomeric protein tagged with DsRed. We demonstrate the feasibility of this approach on the subunit proteins of an oligomeric membrane channel, gap junction connexins. Additionally, DsRed fluorescence was easily detected 12-16 h post transfection, much earlier than previously reported, and could readily be differentiated from co-expressed GFP. Thus, this approach can eliminate the major drawbacks of this highly attractive autofluorescent protein. PMID- 11389890 TI - Do mammalian cells synthesize lipoic acid? Identification of a mouse cDNA encoding a lipoic acid synthase located in mitochondria. AB - Lipoic acid is a coenzyme essential to the activity of enzymes such as pyruvate dehydrogenase, which play important roles in central metabolism. However, neither the enzymes responsible for biosynthesis nor the biosynthetic event of lipoic acid has been reported in mammalian cells. In this study, a mouse mLIP1 cDNA for lipoic acid synthase has been identified. We have shown that the cDNA encodes a lipoic acid synthase by its ability to complement a mutant of Escherichia coli defective in lipoic acid synthase and that mLIP1 is targeted into the mitochondria. These findings suggest that mammalian cells are able to synthesize lipoic acid in mitochondria. PMID- 11389891 TI - Arginine kinase of the flagellate protozoa Trypanosoma cruzi. Regulation of its expression and catalytic activity. AB - In epimastigotes of Trypanosoma cruzi, the etiological agent of Chagas' disease, arginine kinase activity increased continuously during the exponential phase of growth. A correlation between growth rate, enzyme-specific activity and enzyme protein was observed. Arginine kinase-specific activity, expressed as a function of enzyme protein, remains roughly constant up to 18 days of culture. In the whole range of the culture time mRNA levels showed minor changes indicating that the enzyme activity is post-transcriptionally regulated. Arginine kinase could be proposed as a modulator of energetic reserves under starvation stress condition. PMID- 11389892 TI - B cell receptor signaling involves physical and functional association of FAK with Lyn and IgM. AB - B cell receptor (BCR) stimulation induces phosphorylation of a number of proteins, leading to functional activation of B lymphocytes. Focal adhesion kinase (FAK) is a non-receptor protein tyrosine kinase, involved in a variety of signaling pathways. In this study, we show that FAK is tyrosine-phosphorylated and activated following BCR stimulation. We also demonstrate constitutive association of FAK with the Src-family kinase Lyn and with components of the BCR. Association of Lyn with FAK which was not correlated with BCR-induced activation of both kinases, appeared to be mediated via the binding of Lyn to the COOH terminal part of the FAK molecule. Our results indicate that FAK is a component of the BCR complex and that it participates in BCR signaling. PMID- 11389893 TI - Phosphatidylserine delivery to endoplasmic reticulum-derived vesicles of plant cells depends on two biosynthetic pathways. AB - Vesicles formed from endoplasmic reticulum (ER) by a cell-free system of leek cells (Allium porrum) are enriched in phosphatidylserine (PS), especially species containing very long chain fatty acids (VLCFA, at least 20 carbon atoms). In plant cells, PS is formed either by PS synthase or the serine exchange enzyme, although it is not known which pathway(s) contribute(s) to PS delivery in the ER derived vesicles (EV), nor to what extent this occurs. Taking advantage of a cell free system, we have shown that PS enrichment originates mainly from the serine exchange enzyme which is the only pathway that synthesizes the VLCFA-PS species. On the other hand, both enzymes synthesize PS with long chain fatty acids (up to 18 carbon atoms), but these species are given to the EV by PS synthase. PMID- 11389894 TI - Identification of novel mast cell genes by serial analysis of gene expression in cord blood-derived mast cells. AB - The gene expression profile of human cord blood-derived mast cells (MCs) was investigated using serial analysis of gene expression (SAGE). A total of 22914 tags, representing 9181 unique transcripts, were sequenced. By selecting tags that were detected more frequently in MCs than in other tissues, genes characteristic of MCs were enriched. Reverse transcription-PCR and the high density oligonucleotide array hybridization confirmed the validity of our SAGE result. About 70% of the selected genes were previously uncharacterized. Northern blot analysis showed the MC-specific expression of selected genes. This inventory will be useful to identify novel genes with important functions in MCs. PMID- 11389895 TI - Dipeptide synthesis by an isolated adenylate-forming domain of non-ribosomal peptide synthetases (NRPS). AB - A deletion mutant of tyrocidine synthetase 1 (DeltaDeltaTY1), comprising the adenylation domain of TY1 as an independent functional adenylate-forming unit, was used to investigate the ability of the adenylation domain in non-ribosomal peptide synthetases to catalyse peptide bond formation from the aminoacyl adenylate intermediate. The results demonstrate that only one substrate amino acid needs to be activated as an aminoacyl adenylate. In view of the potential exploitation of peptide synthetases for enzymatic synthesis of dipeptides of choice, it is important to note that this does not necessarily require a dimodular construct or an intermediate acyl transfer step. PMID- 11389896 TI - Shy1p occurs in a high molecular weight complex and is required for efficient assembly of cytochrome c oxidase in yeast. AB - Surf1p is a protein involved in the assembly of mitochondrial respiratory chain complexes. However its exact role in this process remains to be elucidated. We studied SHY1, the yeast homologue of SURF1, with an aim to obtain a better understanding of the molecular pathogenesis of cytochrome c oxidase (COX) deficiency in SURF1 mutant cells from Leigh syndrome patients. Assembly of COX was analysed in a shy1 null mutant strain by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). Steady-state levels of the enzyme were found to be strongly reduced, the total amount of assembled complex being approximately 30% of control. The presence of a significant amount of holo-COX in the SHY1 disruptant strain suggests that Shy1p may either facilitate assembly of the enzyme, or increase its stability. However, our observations, based on 2D-PAGE analysis of mitochondria labelled in vitro, now provide the first direct evidence that COX assembly is impaired in a Deltashy1 strain. COX enzyme assembled in the absence of Shy1p appears to be structurally and enzymically normal. The in vitro labelling studies additionally indicate that mitochondrial translation is significantly increased in the shy1 null mutant strain, possibly reflecting a compensatory mechanism for reduced respiratory capacity. Protein interactions of both Shy1p and Surf1p are implied by their appearance in a high molecular weight complex of about 250 kDa, as shown by 2D-PAGE. PMID- 11389897 TI - Complementation of bacterial SecE by a chloroplastic homologue. AB - The SecE protein is an essential component of the SecAYE-translocase, which mediates protein translocation across the cytoplasmic membrane in bacteria. In the thylakoid membranes of chloroplasts, a protein homologous to SecE, chloroplastic (cp) SecE, has been identified. However, the functional role of cpSecE has not been established experimentally. In this report we show that cpSecE in cells depleted for bacterial SecE (i) supports growth, (ii) stabilizes, just like bacterial SecE, the Sec-translocase core component SecY, and (iii) supports Sec-dependent protein translocation. This indicates that cpSecE can functionally replace bacterial SecE in vivo, and strongly suggests that the thylakoid membrane contains a SecAYE-like translocase with functional and structural similarities to the bacterial complex. This study further underscores the evolutionary link between chloroplasts and bacteria. PMID- 11389898 TI - Deacylation of the transmembrane domains of Sindbis virus envelope glycoproteins E1 and E2 does not affect low-pH-induced viral membrane fusion activity. AB - The envelope glycoproteins E1 and E2 of Sindbis virus are palmitoylated at cysteine residues within their transmembrane domains (E1 at position 430, and E2 at positions 388 and 390). Here, we investigated the in vitro membrane fusion activity of Sindbis virus variants (derived from the Toto 1101 infectious clone), in which the E1 C430 and/or E2 C388/390 residues had been substituted for alanines. Both the E1 and E2 mutant viruses, as well as a triple mutant virus, fused with liposomes in a strictly low-pH-dependent manner, the fusion characteristics being indistinguishable from those of the parent Toto 1101 virus. These results demonstrate that acylation of the transmembrane domain of Sindbis virus E1 and E2 is not required for expression of viral membrane fusion activity. PMID- 11389899 TI - Binding and regulation of HIF-1alpha by a subunit of the proteasome complex, PSMA7. AB - The hypoxia-inducible factor-1alpha (HIF-1alpha) is an important transcription factor for cellular responses to oxygen tension. It is rapidly degraded under normoxic conditions by the ubiquitin-dependent proteasome pathway. Here we report a critical role of the 20S proteasome subunit PSMA7 in HIF-1alpha regulation. PSMA7 was found to interact specifically with two subdomains of HIF-1alpha. PSMA7 inhibited the transactivation function of HIF-1alpha under both normoxic and hypoxia-mimicking conditions. In addition, we show that the PSMA7-mediated regulation of HIF-1alpha activity is associated with the proteasome pathway. PMID- 11389900 TI - The shapes and sizes of two domains of tropomodulin, the P-end-capping protein of actin-tropomyosin. AB - Tropomodulin, the P-end (slow-growing end)-capping protein of the actin tropomyosin filament, and its fragment (C20) of the C-terminal half were studied by synchrotron small-angle X-ray scattering, restoring low-resolution shapes using an ab initio shape-determining procedure. Tropomodulin is elongated (115 A long) and consists of two domains, one of 65 A in length and the other being similar to C20 in shape and size if the long axes of the two are tilted by about 40 degrees relative to each other. We propose a model for tropomodulin in association with tropomyosin and actin: the N-terminal half of tropomodulin, a rod, binds to the N-terminus of tropomyosin and the C-terminal triangle domain protrudes from the P-end being slightly bent towards the actin subunit at the end, thereby blocking the P-end. PMID- 11389901 TI - Mechanical unfolding of single filamin A (ABP-280) molecules detected by atomic force microscopy. AB - Filamin A (ABP-280), which is an actin-binding protein of 560 kDa as a dimer, can, together with actin filaments, produce an isotropic cross-linked three dimensional network (actin/filamin A gel) that plays an important role in mechanical responses of cells in processes such as maintenance of membrane stability and translational locomotion. In this study, we investigated the mechanical properties of single filamin A molecules using atomic force microscopy. In force-extension curves, we observed sawtooth patterns corresponding to the unfolding of individual immunoglobulin (Ig)-fold domains of filamin A. At a pulling speed of 0.37 microm/s, the unfolding interval was sharply distributed around 30 nm, while the unfolding force ranged from 50 to 220 pN. This wide distribution of the unfolding force can be explained by variation in values of activation energy and the width of activation barrier of 24 Ig-fold domains of the filamin A at the unfolding transition. This unfolding can endow filamin A with great extensibility. The refolding of the unfolded chain of filamin A occurred when the force applied to the protein was reduced to near zero, indicating that its unfolding is reversible. Based on these results, we discuss here the physiological implications of the mechanical properties of single filamin A molecules. PMID- 11389902 TI - Membrane association and conformational change of palmitoylated G(o)alpha. AB - Bovine brain G(o)alpha was specifically palmitoylated in vitro. The apparent dissociation constant for depalmitoylated G(o)alpha (dG(o)alpha) was 0.273 microM, while that for palmitoylated G(o)alpha (pG(o)alpha) was 5.77 nM. The dissociation rate constant (K(21)) and dissociation half-life for dG(o)alpha were 8.4x10(-4) min and 825 min respectively, while no significant dissociation of pG(o)alpha was detected. The limiting membrane insertion pressures for pG(o)alpha and dG(o)alpha were 44.4 mN/m and 41.3 mN/m respectively. These data suggested that palmitoylation facilitated the membrane association of G(o)alpha. Conformational changes of dG(o)alpha and pG(o)alpha detected by monitoring fluorescence spectra and fluorescence quenching were significantly different after they were associated with the membrane. It was suggested that conformational changes of G(o)alpha upon membrane association might be related to regulation of G(o)alpha signaling by palmitoylation. PMID- 11389903 TI - Catalytic activity of ADAM28. AB - ADAMs are membrane-anchored glycoproteins containing a disintegrin and metalloprotease domain that have important roles in fertilization, development, and diseases such as Alzheimer's dementia. Here we present the first evidence for catalytic activity of ADAM28, a protein that is highly expressed in the epididymis and lymphocytes. Recombinant ADAM28 cleaves myelin basic protein at two sites. The catalytic activity of ADAM28 is not sensitive to tissue inhibitors of metalloproteases 1 and 2, but can be abolished by a mutation in the catalytic site. Catalytically active ADAM28 will be valuable for further studies of its role in sperm maturation and lymphocyte function. PMID- 11389906 TI - Global gene expression during short-term ethanol stress in Saccharomyces cerevisiae. AB - DNA microarrays were used to investigate the expression profile of yeast genes in response to ethanol. Up to 3.1% of the genes encoded in the yeast genome were up regulated by at least a factor of three after 30 min ethanol stress (7% v/v). Concomitantly, 3.2% of the genes were down-regulated by a factor of three. Of the genes up-regulated in response to ethanol 49.4% belong to the environmental stress response and 14.2% belong to the stress gene family. Our data show that in addition to the previously identified ethanol-induced genes, a very large number of genes involved in ionic homeostasis, heat protection, trehalose synthesis and antioxidant defence also respond to ethanol stress. It appears that a large number of the up-regulated genes are involved in energy metabolism. Thus, 'management' of the energy pool (especially ATP) seems to constitute an ethanol stress response and to involve different mechanisms. PMID- 11389904 TI - A discrete amino terminal domain of Kv1.5 and Kv1.4 potassium channels interacts with the spectrin repeats of alpha-actinin-2. AB - The interaction between the amino terminus of Kv1-type potassium channels and alpha-actinin-2 has been investigated. Using a combination of yeast two-hybrid analysis and in vitro binding assays, alpha-actinin-2 was found to bind to the N termini of both Kv1.4 and Kv1.5 but not to the equivalent segments of Kv1.1, Kv1.2 or Kv1.3. Deletion analysis in the in vitro binding assays delineated the actinin binding region of Kv1.5 to between amino acids 73 and 148 of the channel. The Kv1.5 binding sites in alpha-actinin-2 were found to lie within actinin's internal spectrin repeats. Unlike the reported interaction between actinin and the NMDA receptor, calmodulin was found to have no effect on actinin binding to Kv1.5. PMID- 11389905 TI - Functional redundancy of the zinc fingers of A20 for inhibition of NF-kappaB activation and protein-protein interactions. AB - The tumor necrosis factor (TNF) inducible protein A20 is a potent inhibitor of nuclear factor-kappaB (IkappaB)-mediated gene expression in response to TNF and several other stimuli. The C-terminal domain of A20 is characterized by seven zinc finger structures. Here, we show that a minimum of four zinc fingers is required to inhibit TNF-induced nuclear factor-kappaB (NF-kappaB) activation to a level that is comparable to that obtained with the wild-type A20 protein. However, there was no strict requirement for a particular zinc finger structure, since a mutant A20 protein containing only the first four zinc fingers was as potent as a mutant protein containing only the last four zinc fingers. A similar functional redundancy of the A20 zinc fingers was also observed for binding of A20 to a number of other proteins, including two novel NF-kappaB inhibitory proteins (ABIN-1, ABIN-2), A20 itself, the anti-apoptotic protein TXBP151, and a regulatory component of the IkappaB kinase complex, IKKgamma. Moreover, we demonstrate that complete loss of binding of any of these proteins correlates with complete loss of A20's ability to inhibit TNF-induced NF-kappaB activation. However, binding of IKKgamma as such is not sufficient for inhibition of NF kappaB dependent gene expression in response to TNF. PMID- 11389907 TI - Functional consequences of the autosomal dominant G272A mutation in the human GLUT1 gene. AB - The first autosomal dominant missense mutation (G272A) reported within the human GLUT1 gene and shared by three affected family members was investigated in respect to functional consequences. Substitution of glycine-91 by site-directed mutagenesis with either aspartate or alanine resulted in a significant decrease in transport activity of GLUT1 expressed in Xenopus oocytes. Expression of mutant transporters was confirmed by immunoblot, 2-deoxy-glucose uptake and confocal laser microscopy. The data agree with 3-O-methyl-glucose uptake into patient erythrocytes and indicate that the loss of glycine rather than a hydrophilic side chain (Gly91Asp) defines the functional consequences of this mutation. PMID- 11389908 TI - The predicted beta12-beta13 loop is important for inhibition of PP2Acalpha by the antitumor drug fostriecin. AB - The potential anticancer agent fostriecin (FOS) is a potent inhibitor of the protein Ser/Thr phosphatases PP2A and PP4 and a weaker inhibitor of PP1. Random mutagenesis and automated screening in yeast identified residues in human PP2Acalpha important for inhibitory FOS binding. A C269S substitution in the predicted beta12-beta13 loop decreased the FOS sensitivity of intact cells and increased the IC(50) of PP2Acalpha by 10-fold in vitro. Changing PP2Acalpha Cys 269 to phenylalanine, the equivalent residue in PP1, and the Y267G and G270D substitutions caused a similar effect. The results provide information relevant to the design of novel protein Ser/Thr phosphatase inhibitory drugs. PMID- 11389911 TI - Holocene vegetational and coastal environmental changes from the Lago Crispim record in northeastern Para State, eastern Amazonia. AB - Vegetational and coastal environmental changes have been interpreted from a 600cm long and 764014C yr B.P. old sediment core from Lago Crispim located in the northeastern Para State in northern Brazil. The radiocarbon dated sediment core was studied by multi-element geochemistry, pollen and charcoal analysis.Holocene Atlantic sea-level rise caused an elevation of local water table, which allowed the formation of organic deposits in a probably former inter-dune valley. Dense, diverse and tall Amazon rain forest, and some restinga (coastal vegetation) covered the study area at the beginning of the record at 764014C yr B.P. Mangrove vegetation developed along rivers close to the core site at that time. Subsequent decrease in less mangrove vegetation near the study site indicates a sea-level regression, beginning since around 700014C yr B.P. Lower sea-levels probably favoured the formation of a local Mauritia/Mauritiella palm swamp at 662014C yr B.P. Oscillations of higher and lower sea-level stands probably changed the size of the local palm swamp area several times between 6620 and 363014C yr B.P. Sea level transgression at around 363014C yr B.P., caused marked coastal environmental changes: the development of mangroves near the site, the replacement of the local palm swamp by a Cyperaceae swamp, the substitution of the surrounding former Amazon rain forest and some restinga vegetation mainly by salt marshes. High amount carbonised particles suggest a strong human impact by burning on the coastal ecosystems during this late Holocene period.Highest concentrations of NaCl and also Ca, Mg and K in the upper sediment core indicate that the Atlantic was close during the late Holocene period. The core site, which is today 500m from the coastline and only 1-2m above modern sea-level, was apparently never reached by marine excursions during the Holocene.Less representation of mangrove since ca. 184014C yr B.P., may be related due to a slightly lower sea-level or to human impact in the study region. The modern shallow lake seems to be formed recently by road construction, forming an artificial dam. PMID- 11389909 TI - Binding specificity of siglec7 to disialogangliosides of renal cell carcinoma: possible role of disialogangliosides in tumor progression. AB - Previous studies indicate that expression of higher gangliosides in renal cell carcinoma (RCC) is correlated with metastatic potential, particularly in the lung. Out of five major gangliosides in RCC, three disialogangliosides (disialogalactosylgloboside, IV(3)NeuAcIII(6)NeuAcLc(4), and IV(4)GalNAcIV(3)NeuAcIII(6)NeuAcLc(4)) bind strongly to siglec7, which is expressed highly in monocytes and natural killer cells. Out of other gangliosides tested, 2-->6 sialylparagloboside, GD3, GD2, and GT1b, but not other lacto- or ganglio-series gangliosides, showed clear binding to siglec7. In view of preferential metastasis of RCC to the lung, and binding of RCC cell line TOS-1 to lung tissue sections as shown in our previous study, we examined expression of siglec7 in the lung. siglec7 is expressed highly in resident blood cells, but not in parenchymatous cells. TOS-1 cells aggregate together strongly through adhesion with peripheral blood mononuclear cells to form large clumps. This suggests the possibility that such aggregates may form embolisms of microvasculature, particularly in the lung, which initiate metastasis. Other possible roles of higher gangliosides in RCC in promoting metastasis and tumor progression are discussed. PMID- 11389912 TI - A new species of vascular plants from the Xujiachong Formation (Lower Devonian) of Yunnan Province, China. AB - A new species of early land vascular plants, Huia gracilis, is described from the Lower Devonian (Pragian-early Emsian) Xujiachong Formation, Qujing district, Yunnan Province, southwestern China. The plant possesses K- or H-type branching in the rhizome and a dichotomously branched erect system. An axillary tubercle is sometimes present at the branching position. Fertile axes terminate in loose spikes comprising terminal and lateral sporangia arranged helically. Sporangia with long stalks are ovate or elongate-ovate and reflex adaxially. The sporangium dehisces longitudinally in the radial plane of the fertile axis into two parts. The xylem is probably centrarch. Tracheids of G-type are characterised by annular secondary thickenings, between which is a sheet with irregular simple perforations. There may be some 'pores' in the enclosing wall of the perforation. H. gracilis differs from the type species H. recurvata mainly in the branching pattern, more slender morphology of the stem, sporangium and stalk. It is suggested that Huia may have originated from the primitive Cooksonia-like plants. PMID- 11389913 TI - An Early Permian plant assemblage from the Taiyuan Formation of northern China with compression/impression and permineralized preservation. AB - A small but diverse fossil flora is described from the Early Permian Taiyuan Formation occurring at the Yangshuling mine in Pingquan district of Hebei Province, northern China. Fossils occur as compression/impressions within mudrocks and fine-grained sandstones and also as carbonate permineralizations within volcaniclastic tuffs. All are fragmentary and contain lycopsids, sphenopsids, ferns and seed plants, and include several new species. In the compression assemblage sphenopsid and pteridosperm foliage accounts for the majority of the fossils recognised with only a few other kinds of plant organs present. In contrast, the permineralized assemblage is dominated by cordaitaleans with a composition similar to that occurring in coal-ball assemblages elsewhere in the Taiyuan Formation. From the taxonomic synthesis presented it is apparent that the Yangshuling permineralized assemblage contains many of the plant taxa diagnostic of the northern realm of the Early Permian Cathaysian flora, and preserves a representative sample of the wetland coal-swamp vegetation of this time. The permineralized assemblage at Yangshuling represents the first example of anatomically preserved plants from volcaniclastic lithologies from the Palaeozoic of China, raising the possibility of similarly preserved plant-fossil assemblages elsewhere in the Cathaysian realm. PMID- 11389914 TI - Sun and shade leaves? Cuticle ultrastructure of Jurassic Komlopteris nordenskioeldii (Nathorst) Barbacka. AB - An ultrastructural transmission electron microscope (TEM) study of fossil leaf cuticles from the Jurassic pteridosperm Komlopteris nordenskioeldii (Nathorst) Barbacka from the Mecsek Mountains (South Hungary) was conducted. Remnants of cuticles of leaves originating from so-called "sun and shade" environments were sectioned with a diamond knife, transversally as well as longitudinally. Although the present study showed a simple type of cuticle in this pteridosperm, differences were observed in the occurrence of its components, such as electron lucent amorphous material and various densities of granules, which give rise to different zones. The included fibrilous elements appeared to be made of aggregated and aligned granules, equivalent in size and electron density to nearby non-fibrilous granular regions. The combinations of these ultrastructural features allow distinctions between four types of cuticle: sun upper, sun lower, shade upper and shade lower. Considering the distinction made earlier in two types of cuticle and supposed to be related to sun and shade on the basis of macroscopical and microscopical features, four types only on the basis of differences in thickness, the present study reinforces the distinctions with ultrastructural microcharacteristics. As this study shows the variations in ultrastructure of cuticle among the four types, the differences observed may reveal the great sensitivity of some plants to environment. At the same time, it points out the importance, in ultrastructural studies of cuticles, of studying a number of samples for one taxon. PMID- 11389915 TI - Cuticles of Mariopteris occidentalis White nov. emend. from the Middle Pennsylvanian of Oklahoma (USA), and a new type of climber hook for mariopteroid pteridosperms. AB - Cuticles of Mariopteris occidentalis are described from the Desmoinesian (Middle Pennsylvanian) of Oklahoma (USA). This species, like other mariopteroids, had a vine- to liana-like growth habit and climbed with specialized climber hooks. However, M. occidentalis is different from other mariopteroids in having small recurved hooks on the abaxial surfaces of the pinna axes. The diagnosis for M. occidentalis White 1899 is emended based on additional macroscopical observations and data on the epidermal anatomy; a lectotype is designated. M. occidentalis is compared with Pseudomariopteris cordato-ovata from the Stephanian and Autunian of Europe and North America, a taxon which is considered very similar and may be related. Although the two taxa indeed display similarities, significant evidence for a closer relationship could not be found. Finally, some features, e.g. marginal water pits and the stomatal structure, are considered with regard to their palaeoautecological significance where they are interpreted as adaptations to special physiological requirements of a vine- to liana-like life form. PMID- 11389916 TI - Modern pollen and stomate deposition in lake surface sediments from across the treeline on the Kola Peninsula, Russia. AB - We sampled and analyzed surface sediments from 31 lakes along a latitudinal transect crossing the coniferous treeline on the Kola Peninsula, Russia. The major vegetation zones along the transect were tundra, birch-forest tundra, pine forest tundra, and forest. The results indicate that the major vegetation types in our study area have distinct pollen spectra. Sum-of-squares cluster analysis and principal components analysis (PCA) groupings of pollen sites correspond to the major vegetation zones. PCA ordination of taxa indicates that the first axis separates taxa typical of the forest zone (Pinus, Picea) from taxa typical of tundra and forest-tundra zones (Polypodiaceae, Ericaceae, and Betula). The current position of the coniferous treeline, defined in our region by Pinus sylvestris, occurs roughly where Pinus pollen values reach 35% or greater. Arboreal pollen (AP)/non-arboreal pollen (NAP) ratios were calculated for each site and plotted against geographic distance along the transect. AP/NAP ratios of 7 or greater are found within pine-forest tundra and forest vegetation zones. Pinus stomates (dispersed stomatal guard cells) are absent from sites north of the coniferous treeline and all but two samples from the forested sites contain stomates. Stomate concentrations among the samples are highly variable and range from 10 to 458 per ml and positively correlate with the changing Pinus pollen values. PMID- 11389917 TI - Precipitation signal in pollen rain from tropical forests, South India. AB - We have analyzed the pollen content of 51 surface soil samples collected in tropical evergreen and deciduous forests from the Western Ghats of South India sampled along a west to east gradient of decreasing rainfall (between 11 degrees 30-13 degrees 20'N and 75 degrees 30-76 degrees 30'E). Values of mean annual precipitation (Pann, mm/yr) have been calculated at each of the 51 sampling sites from a great number of meteorological stations in South India, using a method of data interpolation based on artificial neural network. Interpolated values at the pollen sites of Pann range from 1200 to 5555mm/yr, while mean temperature of the coldest month (MTCO) remains >15 degrees C and humidity factor (AET/PET, the actual evapotranspiration to potential evapotranspiration ratio) remains also included between 65 and 72%.Results are presented in the form of percentage pollen diagrams where samples are arranged according to increasing values of annual precipitation. They indicate that the climatic signal of rainfall is clearly evidenced by distinct pollen associations. Numerical analyses show that annual precipitation is an important parameter explaining the modern distribution of pollen taxa in this region. Pollen taxa markers of high rainfall (Pann >2500mm/yr) are Mallotus type, Elaeocarpus, Syzygium type, Olea dioica, Gnetum ula, and Hopea type, associated with Ixora type and Caryota. Pollen taxa markers of low rainfall (Pann <2500mm/yr) are Melastomataceae/Combretaceae, Maytenus type, Lagerstroemia and Grewia. The proportions of evergreen taxa and of arboreal taxa vary according to rainfall values. Indeed, when rainfall is <2500mm/yr, percentage of arboreal pollen (AP) is <50% and proportion of evergreen taxa is <20%. When rainfall exceeds 2500mm/yr, AP values average 70%, and proportion of evergreen taxa increases from 60 to 90%. Moreover, a good correlation between precipitation and proportion of evergreen taxa (0.85) presumes that precipitation can be estimated from pollen data. PMID- 11389918 TI - Late Quaternary records of Najas spp. (Najadaceae) from the southwestern Baltic region. AB - Seeds of the submerged vascular plants Najas marina, Najas minor and Najas flexilis are reported from submarine Holocene deposits from the southwestern part of the Baltic Sea, and we also report on a find of Najas minor from an Eemian deposit in Jutland, which is the first record of this species from the Eemian of Denmark. The common and widespread occurrence of especially the southern extralimital N. minor is indicative of higher than present summer temperatures during the period from 10300 to 8000cal.yearsBP. PMID- 11389919 TI - Erratum to "Palynostratigraphy of the last centuries in Switzerland based on 23 lake and mire deposits: chronostratigraphic pollen markers, regional patterns, and local histories" PMID- 11389920 TI - Array-based gene expression profiling to study aging. AB - With recent sequencing of the genome and development of high-density array technology, it is now possible to assess global gene expression in cells/tissues by a technique that is sensitive, quantitative, and rapid. Gene expression array technology is extremely useful in studying a complex, multigenetic process, such as aging, where one needs to understand the interaction of a large number of genes. Although the technology holds great promise, it is novel and not yet well established and there are no widely-accepted standards to guide investigators in the analysis and interpretation of the data obtained. Gene expression array analysis requires strong biostatistical support to minimize false-positives and maximize true-positives in candidate gene identification. It also requires independent validation of the array measurements using other detection methods. Confirmation that differentially expressed (transcribed) genes are reflected by differential expression at the protein level will ultimately be an important measurement. In this review, we focus on the three steps necessary for aging studies when using the gene expression array technology: (1) array hybridization; (2) biostatistical analysis; and (3) array result confirmation. Genes identified by several investigators for their age-associated change using the gene expression array systems are also discussed. PMID- 11389921 TI - Age-dependent fatigue in single intact fast- and slow fibers from mouse EDL and soleus skeletal muscles. AB - In the present work, we investigate age-dependent changes in isometric endurance in response to repetitive stimulation in single intact fast- and slow-twitch muscle fibers from young and old mice. To examine this issue we performed in vitro experiments in manually dissected EDL and soleus muscle fibers. We examined the force generation capacity of fibers in response to two stimulation protocols characterized by different inter-tetanic intervals, named short (1-s) and long interval (3.65-s). Fatigability was measured according to the fatigue index (FI, ratio between the maximum tension recorded in the last over the first tetanus in a train of pulses), the time course of the FI and sag (gradual decrease in force during a partially fused tetanic contraction). Fibers were classified according to the FI using two different criteria previously used in the literature (first criterion: FI > or = 1, 075-099, 0.5-074 and < 0.5; second criterion: FI > or = 1, 0.75-0.99, 0.25-0.74 and < 0.25). The fatigue index distribution recorded in the population of fibers corresponding to EDL and soleus muscles from young and old mice studied with the short and long interval protocols was not statistically different. In summary, these results support the concept that the decline in mechanical performance with aging is not related with changes in fatigability of individual fast- or slow twitch muscles fibers. PMID- 11389922 TI - Caloric restriction alters the feeding response of key metabolic enzyme genes. AB - Differential 'fuel usage' has been proposed as a mechanism for life-span extension by caloric restriction (CR). Here, we report the effects of CR, initiated after weaning, on metabolic enzyme gene expression 0, 1.5, 5, and 12 h after feeding of 24-month-old mice. Plasma glucose and insulin were reduced by approximately 20 and 80%. Therefore, apparent insulin sensitivity, as judged by the glucose to insulin ratio, increased 3.3-fold in CR mice. Phosphoenolpyruvate carboxykinase mRNA and activity were transiently reduced 1.5 h after feeding, but were 20-100% higher in CR mice at other times. Glucose-6-phosphatase mRNA was induced in CR mice and repressed in control mice before, and for 5 h following feeding. Feeding transiently induced glucokinase mRNA fourfold in control mice, but only slightly in CR mice. Pyruvate kinase and pyruvate dehydrogenase activities were reduced approximately 50% in CR mice at most times. Feeding induced glutaminase mRNA, and carbamyl phosphate synthetase I and glutamine synthase activity (and mRNA). They were each approximately twofold or higher in CR mice. These results indicate that in mice, CR maintains higher rates of gluconeogenesis and protein catabolism, even in the hours after feeding. The data are consistent with the idea that CR continuously promotes the turnover and replacement of extrahepatic proteins. PMID- 11389923 TI - Age related changes of the collagen network of the human heart. AB - The objective of this work was to study the collagen tissue of the human heart muscle as a function of age. The types of collagen, their disposition, as well as the density of collagen tissue and diameter of collagen fibrils were examined. Pieces of the ventricular wall from 12 human hearts, six from young individuals and six from aged individuals were studied by the Picrosirius-polarization method and by transmission electron microscopy (TEM). The results obtained showed the presence of two types of collagen fibers in the ventricular walls - thin, weakly birefringent, greenish fibers (collagen type III) and thick, yellow or red, strongly, birefringent fibers (collagen type I), both in the endomysium and perimysium. In the hearts obtained from old subjects, there were no significant differences in the arrangement of the collagen fibers in relation to the hearts obtained from young subjects. Measurements of collagen content in myocardial tissue suggest that both perimysial and endomysial collagen type I fibers increase in number and thickness in the old. These histochemical results obtained coincided with the electron microscopic observations in showing increase in the number of collagen fibrils with large diameter in the old hearts. These ultrastructural and histochemical characteristics of collagen may provide insights important to assessing the pathogenesis of the cardiac lesions of several cardiopathies in the aged heart. PMID- 11389924 TI - Effect of 17-beta estradiol and epidermal growth factor on DNA and RNA labeling in astroglial cells during development, maturation and differentiation in culture. AB - Growth factors stimulate astroglial and neuronal proliferation and differentiation in culture. Estrogens markedly influence astroglia, and are key factors participating in neurodegeneration. The aim of the present study was to investigate interactions between estradiol (E2) and epidermal growth factor (EGF) during astroglia development, maturation and differentiation in culture. DNA or RNA labeling in 16 or 40 or 60 days in vitro (DIV) astrocyte cultures treated for 24 or 48 h with EGF and/or E2 was evaluated. A significant increase in DNA labeling in 16 DIV astrocyte cultures treated for 24 h with EGF (5 ng/ml) and E2 (1 nM) was found. EGF (5 or 10 ng/ml) addition in the last 24 h in 48 h E2 (1 or 5 nM)-treated astrocyte cultures at 16 DIV caused a slight, but significant increase in DNA labeling. No differences in RNA labeling were observed in 16 DIV astrocyte cultures treated for 24 or 48 h with EGF (5 or 10 ng/ml) in the presence of E(2) (1 or 5 nM). A significant stimulation in DNA labeling was shown in 40 DIV astrocyte cultures treated for 48 h with E2 (1 or 5 nM) in the presence of EGF (5 or 10 ng/ml) added in the last 24 h. In well differentiated astroglial cell cultures (60 DIV), DNA labeling was remarkably increased after 24 h treatment with 1 nM E2 or 5 ng/ml EGF. Co-addition of 1 nM E2 and 5 ng/ml EGF for 24 h reduced [methyl-(3)H]thymidine incorporation, when data are compared to E2- or EGF-treated cultures. Addition of EGF in the presence of E2 for 48 h or only in the last 24 h caused a significant decrease of [methyl-(3)H]thymidine incorporation in comparison with EGF-treated cultures at 60 DIV or with untreated cultures. Treatment of cultures for 24 h with EGF (5 or 10 ng/ml) alone or in combination with E2 (1 or 5 nM) induced a strong increase of RNA labeling in 60 DIV astrocyte cultures. Addition for 48 h of E2 (1 or 5 nM) or EGF (5 or 10 ng/ml) alone or in association stimulated significantly RNA labeling in astrocyte cultures at 60 DIV. When 60 DIV astrocyte cultures were treated for 48 h with E2 (1 or 5 nM) in the presence of EGF (5 or 10 ng/ml) added only in the last 24 h, a potentiating effect of RNA labeling was observed. The above results suggest that interaction between growth factors and estrogens may contribute to regulate astroglia development, maturation and differentiation. PMID- 11389925 TI - Increased calcium vulnerability of senescent cardiac mitochondria: protective role for a mitochondrial potassium channel opener. AB - In senescence, endogenous mechanisms of cardioprotection are apparently attenuated resulting in increased vulnerability to ischemia-reperfusion. In particular, mitochondria, which are essential in maintaining cardiac energetic and ionic homeostasis, are susceptible to Ca2+ overload, a component of metabolic injury. However, effective means of protecting senescent mitochondria are lacking. Here, mitochondrial function and structure were assessed using ion selective mini-electrodes, high-performance liquid chromatography and electron microscopy. Aging decreased ADP-induced oxygen consumption and prolonged the time associated with ADP to ATP conversion, which manifested as a reduced rate of oxidative phosphorylation. Aging also reduced mitochondrial Ca2+ handling, and increased Ca2+-induced mitochondrial damage. Diazoxide, a potassium channel opener, reduced Ca2+ loading and protected the functional and structural integrity of senescent mitochondria from Ca2+-induced injury. In this way, the present study identifies the potential usefulness for pharmacotherapy in protecting vulnerable senescent mitochondria from conditions of Ca2+ overload, such as ischemia-reperfusion. PMID- 11389926 TI - Tailoring cancer vaccines to the elderly: the importance of suitable mouse models. AB - The incidence of cancer has increased over the last decade, mainly due to an increase in the elderly population. Vaccine therapy for cancer is potentially less toxic than chemotherapy or radiation and could, therefore, be especially effective in older, more frail cancer patients. However, it has been shown that older individuals do not respond to vaccine therapy as well as younger adults. This has been attributed to T cell unresponsiveness, a phenomenon also observed in cancer patients per se. Activation of tumor-specific T cells by cancer vaccines might be an approach, especially suitable for elderly patients, to eradicate or to prevent recurrence of tumors after primary treatment. To tailor pre-clinical testing of vaccine therapies to the elderly, it is important to have mouse models in which tumors develop at equivalent time points in their life span, as in humans. Such models are currently not available. This progress report first summarizes the current knowledge of tumor-immunological parameters potentially involved in T cell unresponsiveness in relation to aging in mice and humans. Secondly, it reviews those cancer vaccines that are known for their potential to induce tumor-specific T cell responses. Thirdly, it discusses the usefulness of currently available mouse models for pre-clinical testing of cancer vaccines applicable to the elderly population. Finally, experimental approaches are proposed, as to how to develop mouse models that allow the induction of specific tumors at will at different ages, expressing tumor-specific antigens in an 'immune competent' environment. These mouse models may teach us how to overcome immune deficits in the elderly, thereby facilitating the development of effective and safe cancer vaccines. PMID- 11389927 TI - The DNA helicase activity of yeast Sgs1p is essential for normal lifespan but not for resistance to topoisomerase inhibitors. AB - The Saccharomyces cerevisiae SGS1 gene is a member of the RecQ family of ATP dependent DNA helicases, which includes the human WRN, BLM and RECQ4 genes. Mutations in the WRN gene cause the human premature ageing disorder, Werner's syndrome. Deletion of the SGS1 gene also causes premature ageing in yeast, suggesting that the molecular mechanisms of cellular ageing may be evolutionarily conserved. To investigate the role of the RecQ helicase domain in ageing, a point mutation (SGS1 K(706)-->A) known to eliminate the DNA helicase activity of Sgs1p was constructed. This mutant allele failed to rescue the premature ageing of the sgs1Delta strain, demonstrating that Sgs1p DNA helicase activity is required for a normal lifespan. In contrast, the SGS1 K(706)-->A allele was sufficient to rescue the hypersensitivity of the sgs1Delta strain to topoisomerase inhibitors, but not other genotoxic agents. These findings support the idea that Sgs1p fulfils multiple cellular functions, and that DNA helicase activity is dispensable for some of these (e.g. functional interaction with topoisomerases), but essential for others (e.g. longevity). PMID- 11389928 TI - Analysis of microsatellite instability and hypermutation of immunoglobulin variable genes in Werner syndrome. AB - Werner syndrome (WS) is a human premature aging syndrome, which is associated with high frequencies of neoplasia and genetic instability. We have examined the occurrence of microsatellite instability, which may result from defective mismatch repair, in lymphoblastoid cell lines derived from nine WS patients. Instability was measured at the D2S123 locus by gel analysis of PCR products. Three WS cell lines had 4-13% altered alleles, compared with 0% in the other six lines. The increased frequency of microsatellite instability could not readily be associated with overt cancer or any other known clinical condition in the three patients. To examine whether the WS defect affected the humoral immune system, we measured the hypermutation of immunoglobulin variable genes in peripheral blood cells from the WS patient who donated the cell line with the highest frequency of microsatellite instability. The frequency and pattern of mutation was similar to that from normal individuals, suggesting that the Werner protein is not involved in generating hypermutation. PMID- 11389929 TI - Moderate exercise is associated with enhanced antigen-specific cytokine, but not IgM antibody production in aged mice. AB - It has been suggested that moderate exercise may modulate the immune response in the elderly. We investigated whether moderate exercise had an effect on the immune response to viral infection in both young (2-4 months) and older (16-18 months) male BALB/cJ mice. Exercised (EX) mice ran on a treadmill for 8 weeks at a gradually increasing speed and duration whereas control (CON) mice were only handled briefly during each exercise session and then returned to their cages. Mice were infected with herpes simplex virus type 1 (HSV-1) 24 h post-exercise. Serum IgM anti-HSV antibody, HSV-1 specific Th1/Th2 cytokine production by spleen cells, and cytokine production by alveolar cells were measured 7 days post infection. In the aged mice, exercise was associated with an enhanced production of the HSV-1 specific Th1-associated cytokines, interleukin (IL)-2 and interferon (IFN)-gamma, but had no effect on the Th2-associated cytokine IL-10 or IgM antibody. No effect of exercise was observed in young mice. IL-12 production was not altered by exercise, but aging was associated with altered IL-12 production in a tissue-specific manner. In conclusion, moderate exercise was associated with increased antigen-specific IL-2 and IFN-gamma production in response to viral challenge in older mice. PMID- 11389930 TI - Age-related accumulation of oxidative DNA damage and alterations in levels of p16(INK4a/CDKN2a), p21(WAF1/CIP1/SDI1) and p27(KIP1) in human CD4+ T cell clones in vitro. AB - T cells in vivo have been shown to accumulate DNA damage with age. To investigate the effects of DNA damage on T cell biology we have utilised an in vitro human CD4+ T cell clone model. Levels and types of DNA damage were determined in 11 independent T cell clones as a function of their in vitro lifespan. Increased levels of reactive oxygen species (ROS) induced DNA damage with increasing age were found in all clones analysed using a modified alkaline comet assay. T cell clones underwent apoptosis at the end of their lifespans. There were no consistent changes in the mRNA levels for the cyclin-dependent kinase inhibitors (CKI) p16, p21, and p27 during the clones' lifespans. It appears that the increased levels of ROS induced DNA damage in the T cells is not the major trigger of apoptosis, via the p53/p21 pathway. In addition, at the end of their lifespans, the T cell clones did not display the CKI phenotype reported for senescent cells (an increase in p16 and p21 levels). Thus, while the T cell clones appear sensitive to ROS-induced DNA damage, the molecular mechanisms through which this influences T cell dysfunction with age remains to be elucidated. PMID- 11389931 TI - Impaired activation of AP-1 and altered expression of constituent proteins in rat adrenal during ageing. AB - Oxidative stress appears to be one of the primary factors contributing to an age related decline in steroidogenic response in rat adrenocortical and testicular Leydig cells. In this report we concentrate on age-related changes in the DNA binding activity of the transcription factor AP-1 which is particularly responsive to changes in cellular oxidative conditions: adrenal nuclear extracts from young mature (5 months) and old (24 months) rats treated with, and without, lipopolysaccharide (LPS) were studied. AP-1 binding activity, as measured by electrophoretic mobility shift assays (EMSA), was diminished approximately 70% with age in unstimulated adrenals. Following LPS treatment, AP-1 binding activity increased significantly in the adrenals of both young and old animals; however, the level of AP-1 binding achieved in LPS-stimulated old rats was less than that observed for LPS-stimulated young rats. There was no corresponding change in the binding activity of housekeeping transcription factors SP-1 and OCT-1. To further understand these observations, compositional changes in the members of the AP-1 DNA-binding complex were examined by a super-shift assay and Western blot analysis. In adrenals from old rats, a significant decrease in the amount of Fra2 was noted under basal conditions, whereas, substantial decreases in c-Fos, Jun D and c-Jun were observed in response to LPS treatment. In contrast, basal levels of JunB, an inhibitor of the trans-activating function of c-Jun and repressor of AP-1-dependent transcription, were significantly elevated in adrenals from old rats compared to young rats. Together, these findings suggest that ageing-induced oxidative stress may contribute to impaired functional expression of AP-1 by differentially regulating the steady state levels of AP-1 components. The observed decrease in AP-1 binding activity in ageing adrenals is most likely due to decreased expression of the AP-1 activating components (c-Fos, c-Jun, JunD, etc.) and increased expression of JunB, resulting in a switch from transcriptionally active AP-1 complexes observed in young rats to less efficient JunB containing complexes in old rats. PMID- 11389932 TI - Mitochondrial mutations differentially affect aging, mutability and anesthetic sensitivity in Caenorhabditis elegans. AB - In the nematode Caenorhabditis elegans, mutations have been previously isolated that affect the activities of Complex I (gas-1) and Complex II (mev-1), two of the five membrane-bound complexes that control electron flow in mitochondrial respiration. We compared the effects of gas-1 and mev-1 mutations on different traits influenced by mitochondrial function. Mutations in Complex I and II both increased sensitivity to free radicals as measured during development and in aging animals. However, gas-1 and mev-1 mutations differentially affected mutability and anesthetic sensitivity. Specifically, gas-1 was anesthetic hypersensitive but not hypermutable while mev-1 was hypermutable but displayed normal responses to anesthetics. These results indicate that Complexes I and II may differ in their effects on behavior and development, and are consistent with the wide variation in phenotypes that result from mitochondrial changes in other organisms. PMID- 11389933 TI - Impaired migration, integrin function, and actin cytoskeletal organization in dermal fibroblasts from a subset of aged human donors. AB - Deficits in the motility of fibroblasts contribute to age-related impairment of wound healing. We analyzed 'young' fibroblasts from four healthy donors 22-30 years old and 'aged' fibroblasts from six healthy donors 81-92 years old for migratory ability on type I collagen, secretion of matrix metalloproteases (MMPs), attachment to matrices and, expression and function of integrin alpha2beta1. Cells from each donor were analyzed separately in each experiment. Whereas migration of young fibroblasts was uniformly robust, three aged lines migrated well and three migrated poorly. Synthesis of MMP1 and TIMP1, but not MMP2 or MMP9, was increased in the aged fibroblasts relative to the young fibroblast lines irrespective of their motility. All lines of young and aged fibroblasts attached to plastic or collagen with similar efficiency. Although young and aged fibroblasts expressed comparable levels of the alpha2 integrin; the lines of aged fibroblasts that were poor migrators exhibited a significant reduction in alpha2beta1 function relative to fibroblasts with normal migratory capacities. Moreover, the lines of aged fibroblasts that exhibited poor migration demonstrated a disordered actin cytoskeleton and a reduced ability to contract collagen gels. In conclusion, aged fibroblasts, unlike young fibroblasts, displayed variable migratory capacities. Deficient migration by specific lines of aged fibroblasts was not related to the capacity to attach, express alpha2 integrin, or secrete MMPs and TIMP1, but was characterized by disorganized cytoskeletal actin and reduced alpha2beta1 function. PMID- 11389934 TI - The interrelationships of side-chain and main-chain conformations in proteins. AB - The accurate determination of a large number of protein structures by X-ray crystallography makes it possible to conduct a reliable statistical analysis of the distribution of the main-chain and side-chain conformational angles, how these are dependent on residue type, adjacent residue in the sequence, secondary structure, residue-residue interactions and location at the polypeptide chain termini. The interrelationship between the main-chain (phi, psi) and side-chain (chi 1) torsion angles leads to a classification of amino acid residues that simplify the folding alphabet considerably and can be a guide to the design of new proteins or mutational studies. Analyses of residues occurring with disallowed main-chain conformation or with multiple conformations shed some light on why some residues are less favoured in thermophiles. PMID- 11389935 TI - Protein folds propelled by diversity. AB - Many proteins involved in key biological processes are modular in nature. A group of these, the beta-propeller proteins, fold by packing 4-stranded beta-sheets in a circular array. The members of this group are increasingly numerous and, although their modular building blocks all preserve the same basic conformation, they do not have similar sequences. These proteins have extreme functional and phylogenetic diversity. Here, features of the beta-propeller fold are reviewed through comparisons of available structural coordinates. Structure-based sequence alignments combined with analyses of superpositions of individual modular units reveal conserved general features such as hydrogen bonds, beta-turns and positions of hydrophobic contacts. The lack of significant sequence identity is compensated by sets of interactions which stabilise the fold differently in distinct structures. Re-occurring aspartates make contacts to exposed backbone amides in turns or peptide connections within the same sheet. The sole factor responsible for the number of sheets that assemble in the array is the size of the hydrophobic residues that pack into the cores between the sheets. Whilst there is no overall sequence conservation, it may be possible to detect new members of this fold through sequence searches that take into account the repeated nature of the modular assembly as well as the positions of hydrophobic residues and H-bonding side chains. PMID- 11389936 TI - Failure in identification of overlapping spikes from multiple neuron activity causes artificial correlations. AB - Recording of multiple neurons from a single electrode is common practice during extra-cellular recordings. Separation and sorting of spikes originating from the different neurons can be performed either on-line or off-line using multiple methods for pattern matching. However, all spike sorting techniques fail either fully or partially in identifying spikes from multiple neurons when they overlap due to occurrence within a short time interval. This failure, that we termed the 'shadowing effect', causes the well-known phenomenon of decreased cross correlation at zero offset. However, the shadowing effect also causes other artifacts in the auto and cross-correlation of the recorded neurons. These artifacts are significant mainly in brain areas with high firing rate or increased firing synchrony leading to a high probability of spike overlap. Cross correlation of cells recorded from the same electrodes tends to reflect the autocorrelation functions of the two cells, even when there are no functional interactions between the cells. Therefore, the cross-correlation function tends to have a short-term (about the length of the refractory period) peak. A long term (hundreds of milliseconds to a few seconds) trough in the cross-correlation can be seen in cells with bursting and pausing activities recorded from the same electrode. Even the autocorrelation functions of the recorded neurons feature firing properties of other neurons recorded from the same electrode. Examples of these effects are given from our recordings in the globus pallidus of behaving primates and from the literature. Results of simulations of independent simple model neurons exhibit the same properties as the recorded neurons. The effect is analyzed and can be estimated to enable better evaluation of the underlying firing patterns and the actual synchronization of neighboring neurons recorded by a single electrode. PMID- 11389937 TI - Culture and regeneration of human neurons after brain surgery. AB - Cortical human brain tissue was obtained from 11 craniotomies for intractable epilepsy or tumor resection. Neuregen transport medium preserved viability at 4 degrees C during transfer to the culture laboratory. Cells were isolated and cultured by methods previously developed for adult rat neurons (Brewer GJ. Isolation and culture of adult rat hippocampal neurons. J. Neurosci. Meth. 1997:71:143-55). In about 40% of the cases, cultures regenerated with a majority of neuron-like cells that stained for neurofilament and not GFAP. After 3 weeks of culture from a 70 year old meningioma case, synapse-like structures were revealed by electron microscopy. Trophic support from basic human recombinant fibroblast growth factor was synergistically improved with the steroid hormone dehydroepiandrosterone 3-sulfate. Another 40% of the cases resulted in cultures that were predominantly GFAP positive astroglia. The remaining 20% of the cases did not regenerate cells with neuron-like or glial processes. Three postmortem cases did not regenerate neurites. These methods may aid development of human culture models of epilepsy as well as human pharmacology, toxicology and development of improved methods for brain grafts. PMID- 11389938 TI - Contrast optimization of Macaca mulatta basal ganglia in magnetic resonance images at 4.7 Tesla. AB - To determine whether high field MRI could distinguish among the different regions of the basal ganglia, the brains of two Macaca mulatta monkeys were explored in vivo using a 4.7 T MR imager. Gradient-echo (GE) and spin-echo images were acquired with proton-density, T1 and T2* weightings. Five GE images with increased susceptibility effects were generated using a GESFID sequence, from which T2* maps were also reconstructed. The first echo of the GESFID sequence (TE = 12.6 ms) produced the best contrast-to-noise ratio (C/N) between the pallidum and the putamen, the pallidum and the thalamus, the substantia nigra and the surrounding white matter, and the substantia nigra and the subthalamic nucleus. An increased T2*-weighting (TE = 37.2 ms) was necessary to maximize C/N between the putamen and the surrounding white matter, and between the subthalamic nucleus and the surrounding white matter. A dual GE sequence with a short TE ( approximately 10 ms) and a longer one ( approximately 30 ms) thus effectively localizes basal ganglia subregions at 4.7 T. PMID- 11389939 TI - Ex vivo culture of isolated zebrafish whole brain. AB - We have succeeded in culturing whole zebrafish brains ex vivo for 1 week. While isolated cells and tissue slices have previously been employed for neurobiological studies, these techniques are limited, because while local networks may be preserved, their original context in the whole brain is lost. Culture of the whole brain would facilitate the study of cells and systems within an intact brain infrastructure. Our culture method entailed isolating the whole brain and placing it on a sterile and porous membrane, after which it was maintained with a conditioned medium in a six-well plate in a CO2 incubator at 28.5 degrees C. Whole brains cultured by this simple method were relatively unaltered in terms of their morphology, cytoarchitecture, immunohistochemistry and ability to transport horse radish peroxidase (HRP). This method of cultivation may be very useful for neurobiological research. PMID- 11389941 TI - The dispersed cell culture as model for functional studies of the subcommissural organ: preparation and characterization of the culture system. AB - The subcommissural organ (SCO) is an enigmatic secretory gland of the brain, which is believed to be derived from ependymal (glial) precursor cells. We here developed a dispersed cell culture system of the bovine SCO as an approach to functional analyses of this brain gland. Tissue of the bovine SCO obtained from the slaughterhouse was papain dissociated either directly after dissection or after preparation of SCO explants. The latter had been maintained for 4-6 weeks in organ culture. The dispersed cells were cultured for up to 14 days and continuously tested for their secretory state by immunostaining of their secretory product. With respect to the morphology of the SCO cells (shape, processes, nucleus), no difference was found between the culture of freshly dissociated SCOs and that of dissociated SCO explants. In all cases, the dissociation caused a dedifferentiation; typical elongated cells were formed increasingly after 1 day of culture. Thereafter, only the cellular size increased, whereas the shape and the viability of the cells remained unchanged. Proliferating SCO cells were never observed. The culture obtained from fresh SCO tissue contained more glia cells and fibrocytes than the culture prepared from SCO explants. The proliferation of glia cells and fibrocytes was suppressed by blocking the mitotic activity with cytosine-beta-D-arabino furanoside (CAF). The cytophysiological features of the cultured dispersed cells of both origins did not differ as demonstrated by classical histology, by immunocytochemistry for the secretory products of the SCO, by the characteristics of calcium influx into the cytoplasm ([Ca2+]i) and cyclic adenosine monophosphate (cAMP) after stimulation with adenosine-5-triphosphate, substance P or serotonin, and by the activation of the transcription factor cAMP-responsive element-binding protein. Because of the maintenance of their viability, their capacity to release the secretory product into the culture medium, their receptive capacity, and their signal transduction pathways, we conclude that the dispersed cell culture system, especially that obtained from SCO explants, represents an appropriate and useful model for functional studies of the mammalian SCO. PMID- 11389940 TI - A double-labeled preparation for simultaneous measurement of [3H]-noradrenaline and [14C]-glutamic acid exocytosis from streptolysin-O (SLO)-perforated synaptosomes. AB - We have developed a novel method to examine [3H]-noradrenaline and [14C] glutamate release from the same sample of streptolysin-O (SLO) perforated rat cortical synaptosomes. Ca2+ -dependent [3H]-noradrenaline and [14C]-glutamate release was examined at different temperatures and was found to be greater at 30 degrees C than at 25 degrees C. Ca2+ -dependent release of [3H]-noradrenaline is more ATP dependent than Ca2+ -dependent release of [14C]-glutamate. No significant reuptake of either neurotransmitter by the perforated synaptosomes was detected, indicating all the synaptosomes were indeed perforated. Incubations with 1 mM ouabain, a specific Na+,K+ -ATPase inhibitor, slightly increased Ca2+ dependent release of both neurotransmitters. [3H]-noradrenaline is released from large dense-core vesicles and [14C]-glutamate is released from small clear synaptic vesicles, so one can directly compare and contrast neurotransmitter release mechanisms between large dense-core vesicles and small clear synaptic vesicles using this preparation. PMID- 11389942 TI - Evaluation of a stereotactic frame for repositioning of the rat brain in serial positron emission tomography imaging studies. AB - For serial imaging studies of the rat brain with positron emission tomography (PET), reproducible positioning of the head can facilitate spatial alignment of images and quantitative analysis. To achieve this aim, we constructed a plastic head frame and tested the positioning reproducibility on a high-resolution small animal PET scanner, microPET. Two sets of ear bars, with tapers of either 18 degrees (sharp) or 45 degrees (blunt), were evaluated for their relative precision in securing the animal to the frame. For sequential positioning of an animal, average distances from the mean position of 0.51 mm (SD 0.41 mm) and 0.91 mm (SD 0.48 mm) were measured with the sharp and blunt ear bars, respectively. These results show that a rat brain can be reproducibly positioned using the frame, with a variation of position less than the spatial resolution of modern animal PET scanners. Brain regions of interest defined on one scan and copied across subsequent scans of a frame-repositioned animal resulted in an average coefficient of variation of 5.4% (SD 2.7%) using the sharp ear bars and 6.8% (SD 2.5%) using the blunt ear bars. This methodology has the potential to improve quantitative assessment for serial PET studies. PMID- 11389943 TI - Direct comparison of visual cortex activation in human and non-human primates using functional magnetic resonance imaging. AB - We report a technique for functional magnetic resonance imaging (fMRI) in an awake, co-operative, rhesus macaque (Macaca mulatta) in a conventional 1.5T clinical MR scanner, thus accomplishing the first direct comparison of activation in visual cortex between humans and non-human primates with fMRI. Activation was seen in multiple areas of striate and extra-striate visual cortex and in areas for motion, object and face recognition in the monkey and in homologous visual areas in a human volunteer. This article describes T1, T2 and T2* values for macaque cortex, suitable MR imaging sequences, a training schedule, stimulus delivery apparatus and restraining hardware for monkey fMRI using a conventional 19 cm knee coil. Much of our understanding of the functional organization of the primate brain comes from physiological studies in monkeys. Direct comparison between species using fMRI such as those described here will help us to relate the wealth of existing knowledge on the functional organization of the non-human primate brain to human fMRI. PMID- 11389944 TI - A simple method for constructing microinjectrodes for reversible inactivation in behaving monkeys. AB - A method for constructing a simple, durable injection-microelectrode (injectrode) is described. The injectrode can record neuronal activity, stimulate neuronal tissues, or inject substances locally through its tip. The injectrode is lightweight and is easy to construct from commercially available parts, and it can be used repeatedly for multiple recordings and injections. Since dura penetration can damage fragile electrode tips, a reliable method to pass the injectrode through an intact dura matter is described. PMID- 11389945 TI - A nerve stimulation method to selectively recruit smaller motor-units in rat skeletal muscle. AB - Electrical stimulation of peripheral nerve results in a motor-unit recruitment order opposite to that attained by natural neural control, i.e. from large, fast fatiguing to progressively smaller, fatigue-resistant motor-units. Yet animal studies involving physiological exercise protocols of low intensity and long duration require minimal fatigue. The present study sought to apply a nerve stimulation method to selectively recruit smaller motor-units in rat skeletal muscle. Two pulse generators were used, independently supplying short supramaximal cathodal stimulating pulses (0.5 ms) and long subthreshold cathodal inactivating pulses (1.5 s) to the sciatic nerve. Propagation of action potentials was selectively blocked in nerve fibres of different diameter by adjusting the strength of the inactivating current. A tensile-testing machine was used to gauge isometric muscle force of the plantaris and both heads of the gastrocnemius muscle. The order of motor-unit recruitment was estimated from twitch characteristics, i.e. peak force and relaxation time. The results showed prolonged relaxation at lower twitch peak forces as the intensity of the inactivating current increased, indicating a reduction of the number of large motor-units to force production. It is shown that the nerve stimulation method described is effective in mimicking physiological muscle control. PMID- 11389946 TI - The inebri-actometer: a device for measuring the locomotor activity of Drosophila exposed to ethanol vapor. AB - Drosophila melanogaster can be used as a model organism for probing the genetic basis for alcohol sensitivity. In this paper, we describe a new device, the inebri-actometer, which measures the locomotor activity of up to 128 individual flies simultaneously. The device consists of 128 pairs of emitter/detector photodiodes connected in series through a computer interlink. A single fly is placed in each of the 128 chambers and humidified air or air containing variable amounts of ethanol vapor is pumped through the chambers. When a fly blocks the infrared signal transmitted by an emitter photodiode, the computer records one movement for that fly. We present preliminary results showing the effect of ethanol on the activity of wild-type Oregon R Drosophila. Five preliminary runs with 95% ethanol vapor revealed that this concentration induces an approximately 3- to 4-fold increase in locomotor activity which peaks at about 5 min after the addition of ethanol vapor. This is followed by a gradual decrease in activity leading to a nearly total cessation of movement after 30 min. Statistically significant dose-related activity increases were obtained for ethanol concentrations of 8, 19, 50, and 100% of maximum, assessed in two replications at each dose. Unlike the complete suppression of locomotion seen in the last 10 min of the session at maximum ethanol exposure, the initial stimulation effect at the 19% concentration was maintained across the 30-min session. PMID- 11389947 TI - A forebrain ischemic preconditioning model established in C57Black/Crj6 mice. AB - Although many kinds of rat and gerbil cerebral ischemic preconditioning models are available, only a focal ischemic preconditioning model in mice has been reported. As most genetic alterations have been performed in mice, it is urgent to develop mouse ischemic preconditioning models for investigating the molecular mechanisms of ischemic preconditioning in transgenic mice. In the present study, we developed a forebrain ischemic preconditioning model in C57Black/Crj6 (C57BL/6) mice. Forebrain ischemia was induced in C57BL/6 mice (8-10 weeks old) by bilateral common carotid artery occlusion (BCCAO) for 18 min. The conditioning ischemic insult lasting for 6 min was carried out 48 h before the 18-min BCCAO. On the seventh day after BCCAO, neuronal damage was visualized by microtubule associated protein-2 immunohistochemistry and quantified by cresyl violet staining. Terminal deoxytransferase-mediated dUTP-nick end labeling (TUNEL) was performed 72 h after reperfusion to detect DNA fragmentation. Ischemia for 18 min resulted in injury to the striatum, cortex and hippocampus. In comparison to the hippocampus, striatal neuronal injury was more severe and reproducible. Although the conditioning ischemia itself caused neither noticeable striatal neuronal damage nor DNA fragmentation, it significantly reduced striatal neuronal damage and DNA fragmentation caused by the subsequent 18-min ischemia. These results indicate that striatal neuronal injury after transient BCCAO can be strongly reduced by a sublethal ischemic episode in C57BL/6 mice. As many kinds of gene altered C57BL/6 mice are available, this preconditioning model may be useful for investigating the molecular mechanisms of ischemic preconditioning in transgenic mice. PMID- 11389948 TI - A force-plate actometer for quantitating rodent behaviors: illustrative data on locomotion, rotation, spatial patterning, stereotypies, and tremor. AB - This report describes a new kind of actometer for recording the behavior of rodents or other small animals. The instrument, a force-plate actometer, uses a stiff, low-mass horizontal plate coupled to four supporting force transducers positioned at the corners of the plate. When an animal moves on the plate, its movements are sensed by the transducers whose signals are processed by computer to yield measurements of a wide range of behaviors or behavioral attributes, such as locomotor activity, rotation around the center, whole-body tremor, and amphetamine-induced stereotypies. Spatial resolution is less than 1 mm, and temporal resolution is 0.02 s. Sample data were presented comparing the locomotor activity of CD-1, BALB/c, and C57BL/6 mice before and after treatment with D amphetamine sulfate. Rotational behavior was recorded in an amphetamine-treated rat that had sustained a unilateral 6-hydroxydopamine-induced lesion of the nigrostriatal system. In the C57BL/6 mouse, harmaline-induced tremor was quantified. With rats as subjects, the force-plate actometer was used to quantify amphetamine-induced stereotypies, to demonstrate the development of sensitization to amphetamine's effects, and to quantitate the consistent 11-12 Hz rhythmicities that underlie the sterotypies. The performance of the force-plate actometer was compared with that of a variety of instruments reported in the literature on behavioral instrumentation. Finally, potential applications in neuroscience research other than those illustrated in this report were discussed. PMID- 11389949 TI - A simple method for measuring EEG in freely moving guinea pigs. AB - This paper describes a method of measuring EEG in guinea pigs by means of temporarily attached superficial electrodes with radiotelemetry equipment. This represents an alternative approach to more conventional methods of monitoring EEG in animals, which necessitate surgical implantation of electrodes. The novel approach of using superficial electrodes was compared with the cortical screw electrode technique, in a nerve agent-induced seizure model. In both cases, radiotelemetry techniques were used for data capture to facilitate monitoring of nerve agent-induced seizures in freely moving non-tethered guinea pigs. The advantages and disadvantages of the technique are discussed, for example the saving in resources and the benefits to animal welfare. PMID- 11389950 TI - Primary culture of neural precursors from the ovine central nervous system (CNS). AB - The present study demonstrates that bipotential neural precursors isolated from an early developmental stage of the sheep embryo nervous system can be maintained in vitro in an undifferentiated state for a long period. These precursors multiplied under the action of epidermal growth factor and basic fibroblast growth factor and formed free-floating aggregates of nestin-immunoreactive cells, called neurospheres. These precursors can undergo predominantly neural or glial differentiation according to the culture conditions. Medium supplemented with foetal calf serum mainly favoured cell differentiation predominantly into astrocytes, whereas the defined SATO medium favoured neuronal differentiation. Using various immunomarkers of neurones and astroglial cells, we described the course of differentiation of neuronal and astroglial cells in different culture conditions. The ability to grow neural precursors from common laboratory animals has been useful for studying the cellular and molecular mechanisms underlying the development of the central nervous system. Furthermore, neural progenitors are already being used for in vivo cell therapy in various neurodegenerative disorders. The ovine species is a well-known model for prion diseases, since scrapie is endemic in most countries and has been studied for a long time. In this respect, the availability of ovine neural precursors will add a new perspective to the study of the pathogenicity of prion diseases. PMID- 11389951 TI - Detecting precise firing sequences in experimental data. AB - A precise firing sequence (PFS) is defined here as a sequence of three spikes with fixed delays (up to some time accuracy Delta), that repeat excessively. This paper provides guidelines for detecting PFSs, verifying their significance through surrogate spike trains, and identifying existing PFSs. The method is based on constructing a three-fold correlation among spikes, estimating the expected shape of the correlation by smoothing, and detecting points for which the correlations significantly protrude above the expected correlation. Validation is achieved by generating surrogate spike trains in which the time of each of the real spikes is randomly jittered within a small time window. The method is extensively tested through application to simulated spike trains, and the results are illustrated with recordings of single units in the frontal cortex of behaving monkeys. Pitfalls which may cause false detection of PFSs, or loss of existing PFSs, include searching for PFSs in which the same neuron participates more than once, and attempting to produce a surrogate with some fixed statistical property. PMID- 11389952 TI - Regulation of bovine E-selectin expression by recombinant tumor necrosis factor alpha and lipopolysaccharide. AB - Induction of adhesion molecules by cytokines and LPS is an important mechanism of regulating leukocyte migration into tissue. Expression and regulation of E selectin may be differentially influenced by the stimuli involved with effects on mRNA or surface protein kinetics. Surface protein and mRNA expression kinetics of bovine E-selectin were measured and compared in primary cultures of bovine aortic endothelial cells (BAEC) stimulated for various periods of time with recombinant bovine tumor necrosis factor alpha (rbTNF-alpha) or Escherichia coli lipopolysaccharide (LPS). E-selectin mRNA expression was measured via quantitative reverse transcription polymerase chain reaction (Q-RT-PCR) using a construct that contained multiple synthetic oligonucleotides for several bovine adhesion molecules and cytokines. Surface expression of E-selectin was measured by flow cytometry. Unstimulated BAECs expressed minimum or no E-selectin on the surface. A low number of endothelial cells expressed surface E-selectin as early as 1h post-stimulation and surface expression was sustained after both stimuli for 24-72h. Mean fluorescence intensity (MFI) indicated peak surface concentration of E-selectin at 6 h post-stimulation after LPS followed by a gradual decrease to 72h without returning to baseline values. Mean fluorescence intensity following stimulation with TNF-alpha increased slightly between 0 and 72h. The pattern of mRNA expression differed between stimuli. LPS-stimulated BAECs expressed peak amounts of E-selectin mRNA at 6 h, followed by a decline to baseline by 24 h. Conversely, BAECs stimulated with rbTNF-alpha expressed significantly (p pound 0.05) higher amounts of mRNA at 1h than compared to unstimulated controls (0 h), but this decreased to below baseline levels by 6h; followed by a gradual increase and eventually a sharp increase between 18 and 72 h. To account for the lack of correlation between mRNA and protein expression, it was hypothesized that shedding of surface E-selectin accounted at least in part, for the large increase in mRNA expression seen at 18-72h. Culture supernatants from rbTNF-alpha-treated BAECs were harvested, and tested for the presence of shed E-selectin using ELISA. Unstimulated culture supernatants contained little or no E-selectin. Between 6 and 48 h, the concentration of E-selectin in culture supernatants from rbTNF-alpha-stimulated BAECs increased approximately two-fold, suggesting that the sharp increase in E-selectin mRNA expression around 18 h may be related to significant loss of surface E-selectin during this period. PMID- 11389953 TI - Effect of oral administration of high vitamin C and E dosages on the gilthead seabream (Sparus aurata L.) innate immune system. AB - The effect of the oral administration of high dosages of vitamins C and E on the innate immune system of the seabream was investigated. Gilthead seabream (Sparus aurata L.) were fed four different diets for 45 days: a commercial diet as control, a 3g/kg vitamin C-supplemented diet, a 1.2g/kg vitamin E-supplemented diet or a diet containing both vitamin supplements. After 15, 30 and 45 days, serum ascorbic acid and alpha-tocopherol levels, growth, complement titers and head-kidney leucocyte phagocytic and respiratory burst activities were evaluated. The results showed that serum vitamin levels reflected dietary input. Fish fed the vitamin C-supplemented diet showed an enhanced respiratory burst activity, while fish fed the vitamin E-supplemented diet exhibited increased complement and phagocytic activities. All of these positive effects were found in fish fed the joint diet, as well as a synergistically enhanced respiratory burst activity at day 30. The results demonstrate that in vivo vitamins C and E exert a synergistic effect enhancing the respiratory burst activity of seabream phagocytes. PMID- 11389954 TI - Molecular cloning of canine IL-13 receptor alpha chain (alpha1 and alpha2) cDNAs and detection of corresponding mRNAs in canine tissues. AB - This communication reports the cloning of cDNAs encoding two canine IL-13 receptor alpha chains (caIL-13Ralpha1 and caIL-13Ralpha2). As described for the members of type-I cytokine receptors, both caIL-13Ralpha1 and caIL-13Ralpha2 were found to contain the highly conserved motifs, such as cysteine and tryptophan residues in their N-terminal portion and the WSXWS at C-terminus. The isolated caIL-13Ralpha1 cDNA contains 1547 nucleotides with an open reading frame that encodes 405 amino acid residues. Canine IL-13Ralpha1 is 82.0 and 69.3% identical to human and mouse IL-13Ralpha1s, respectively, at the amino acid level. Canine IL-13Ralpha1 has an almost identical cytoplasmic domain to its human and mouse counterparts. The isolated caIL-13Ralpha2 cDNA contains 1454 nucleotides and encodes an open reading frame of 386 amino acid residues. Canine IL-13Ralpha2 is 62.6 and 47.5% identical to its human and mouse counterparts, respectively, at the amino acid level. Using RT-PCR with caIL-13Ralpha1 and caIL-13Ralpha2 specific primers, mRNAs of caIL-13Ralpha1 and caIL-13Ralpha2 were detected in most dog tissues. In addition, RT-PCR detected caIL-13Ralpha1 mRNA in one of two canine mastocytoma (C2 but not Br) cell lines and in a canine macrophage-derived cell line (DH82). CaIL-13Ralpha2 mRNA was detected in all three canine cell lines. PMID- 11389955 TI - Immunological responses of Eurasian badgers (Meles meles) vaccinated with Mycobacterium bovis BCG (bacillus calmette guerin). AB - Wildlife species, such as the badger (Meles meles), may act as maintenance hosts for Mycobacterium bovis and contribute to the spread and persistence of tuberculosis in associated cattle populations. Targeted vaccination of badgers against tuberculosis is an option that, if successfully employed, could directly facilitate the advancement of bovine tuberculosis eradication in affected areas. In this study, the immunological responses of a group of badgers vaccinated subcutaneously with low doses of Mycobacterium bovis bacillus calmette guerin (BCG) were measured in vitro and compared with non-vaccinated control animals over a period of 42 weeks. Peripheral blood mononuclear cells (PBMC) from badgers which had received repeated booster injections of BCG proliferated in response to culture with PPD-bovine (purified protein derivative of tuberculin). The proliferation was significantly greater than that seen in the non-vaccinated control group. In contrast, the proliferative response of PBMC from vaccinated badgers to PPD-avian declined relative to the control group. These results demonstrate that repeated vaccination of badgers with M. bovis BCG induced a population of T-lymphocytes responsive to specific antigens in PPD-bovine. Throughout the course of the study, the sera from all animals were tested (BrockTest) by an enzyme-linked immunosorbent assay (ELISA) system for the presence of antibodies to MPB83, a serodominant antigen whose expression is high in M. bovis, but very low in BCG (Pasteur). No animals at any stage showed seroconversion to the antigen, consistent with the tuberculosis-free status of the badgers under study. PMID- 11389956 TI - Molecular cloning of feline interferon-gamma-inducing factor (interleukin-18) and its expression in various tissues. AB - Interleukin-18 (IL-18) is a cytokine with potent interferon-gamma-inducing activity, and plays an important biologic role in the enhancement of the activity of natural killer cells and cytotoxic T-lymphocytes. In this study, feline IL-18 cDNA was cloned and characterized to establish a basis for the prospective cytokine therapy in small animal practice. The nucleotide sequence of feline IL 18 cDNA obtained in this study was 712bp long and contained its entire open reading frame encoding 192 amino acid residues. The predicted amino acid sequence of feline IL-18 cDNA showed 77.2, 84.8, 60.2 and 62.6% similarity with those of human, dog, rat and mouse counterparts, respectively. The feline IL-18 cDNA included a putative cleavage site of IL-1beta-converting enzyme (ICE) and IL-1 signature-like sequences identified in human and mouse IL-18 cDNAs. Expression of IL-18 mRNA was detected in various tissues including spleen, liver and cerebrum in the cat. PMID- 11389957 TI - Characterization of the innate and adaptive immunity to Salmonella enteritidis PT1 infection in four broiler lines. AB - Four broiler lines were inoculated orally with Salmonella enteritidis phage type 1 at the age of 7 days (experiment A: lines 1 and 2) and at the age of 1 day (experiment B: lines 3 and 4). At various days post-infection chickens were sacrificed and the number of Salmonella in the caeca, liver, and spleen were determined. Furthermore, phagocytic activity, cellular immune responses, and humoral responses were determined using, respectively, single-cell suspensions of spleen or intestine and serum. In both experiments, similar trends were seen. Increased numbers of S. enteritidis were found in the caeca of lines 1 and 3, whereas at the same time a decreased colonization was found in the spleen and in the liver, as compared to lines 2 and 4. In the latter two lines, the phagocytic activity of the phagocytes was higher and the humoral responses were lower. Observations from this study suggest that lower activity of phagocytes and higher humoral activity prevent systemic S. enteritidis infection. PMID- 11389958 TI - Functional restoration of the bursa of Fabricius following in ovo infectious bursal disease vaccination. AB - The primary role of the avian bursa of Fabricius is to provide an essential microenvironment for B-lymphocytes to diversify their immunoglobulin genes by gene hyperconversion. Infectious bursal disease (IBD) vaccination using intermediate plus vaccine strains can temporarily deplete the bursal follicles and interrupt the normal B-cell development, which is generally followed by B cell repopulation and histological regeneration. To find evidence that functional restoration of the bursa of Fabricius occurs in addition to the histological regeneration, we have analysed the chB1 gene expression, which indicates active bursal B-lymphocytes, and also the surface expression of a carbohydrate structure Lewis(x), a marker which identifies those bursal B-lymphocytes that are undergoing gene hyperconversion. In ovo vaccination with an immune complex vaccine (IBDV-BDA) caused transient bursal destruction in both the SPF and the maternally protected broiler groups with differences evident in the starting time, the severity and the duration of the effect. After the depletion phase, signs of histological regeneration appeared together with chB1- and Lewis(x) expression indicating that B-lymphocytes were functionally active and the bursa of Fabricius was serving again as an efficient primary lymphoid organ providing an appropriate microenvironment for B-cell development. PMID- 11389959 TI - Comparative evaluation of the CD4+CD8+ and CD4+CD8- lymphocytes in the immune response to porcine rubulavirus. AB - The porcine immune system is unique in the expression of CD4+CD8+ (double positive, DP) lymphocytes. These cells have been associated with immunological memory due to their gradual increase with age, the expression of memory phenotype and their ability to respond to recall viral antigen. This work analyzes the biological function of CD4+CD8- and CD4+CD8+ lymphocytes in the immune response to porcine rubulavirus (PRv). CD4+CD8- cells isolated from pigs 3 weeks after infection with porcine rubulavirus proliferated in response to homologous virus and generated lymphoblasts which were predominantly of the CD4+CD8+ phenotype, whereas stimulation with mitogen induced proliferation but did not switch the phenotype. CD4+CD8- lymphocytes isolated after 10 weeks of infection proliferated in response to phytohemagglutinin (PHA) but did not proliferate in response to homologous virus and did not change their phenotype, whereas CD4+CD8+ lymphocytes proliferated in response to PHA and to viral antigen. The cytokine profile of both lymphocyte populations showed the presence of IL-2 and IL-10 transcripts, quantitation demonstrated that CD4+CD8+ cells expressed mainly IL-10, whereas CD4+CD8- lymphocytes expressed primarily IL-2. Our results show that CD4+CD8- lymphocytes in the early phase of porcine rubulavirus infection can be converted to double-positive cells expressing IL-10 in an antigen-dependent manner, and that CD4+CD8- T-cells late in infection do not acquire CD8. PMID- 11389960 TI - Densitometric analysis of Western blot assays for feline immunodeficiency virus antibodies. AB - Western blot (WB) strips for antibodies directed to feline immunodeficiency virus (FIV) were analysed using reflectance densitometry by a semiautomatic densitometer. This method was used to quantify the antibody responses to different FIV proteins in both vaccinated and naturally or experimentally infected cats. In order to increase reproducibility, reagents and protocols were accurately standardised and internal controls were added. In a first format, an internal control band consisting of feline IgG was added to each blot to minimise the effect of band intensity variation. In a second format, antibody concentrations were calculated from the ratio of the densities produced by test sera and by positive and negative standard sera. The sera under scrutiny were also examined by standard enzyme-linked immunosorbent assay (ELISA) and the results obtained compared with those of the corresponding WB. A statistically significant positive correlation was found between the results obtained with the two methods, and this was especially evident when ELISA titres were compared to corrected WB values (P = 0.001). Densitometric analysis of WB assays allowed to quantify the antibodies against FIV proteins and might be useful to investigate possible humoral immune correlates of protection in FIV vaccination studies and antibody production in the early phase of infection. The quantitation of antibodies to Gag and Env FIV antigens might be used to obtain further informations on the course of FIV disease, as previously demonstrated in human immunodeficiency virus-1 (HIV-1) infections. PMID- 11389961 TI - Cell mediated immunity and specific IgG1 and IgG2 antibody response in natural and experimental canine leishmaniosis. AB - In the present study, we have followed up Leishmania infantum infection in dogs: (1) naturally infected; (2) experimentally infected with amastigotes; and (3) experimentally infected with culture promastigotes. The main objective was to evaluate the differences of the humoral and cellular immune responses of each group. Sera from 12 beagle dogs were analysed for total anti-leishmanial antibodies and IgG1 and IgG2 subclasses by enzyme-linked immunosorbent assay (ELISA). Lymphoproliferation to L. infantum antigen was also performed. All naturally infected animals were symptomatic with a marked humoral response. Dogs inoculated with amastigotes were asymptomotic and presented lower antibody titres than naturally infected. Dogs inoculated with culture promastigotes were asymptomotic with no significant humoral response. Strong proliferative responses to Leishmania antigen was observed in dogs inoculated with promastigotes. In our experimental model, IgG1 antibody levels presented a similar pattern in all infected animals, and IgG2 reactivity was high in naturally infected dogs. PMID- 11389963 TI - A novel and highly divergent Arabidopsis cyclin isolated by complementation in budding yeast. AB - A novel cyclin, CycJ18, was isolated by complementation of G1 cyclin-deficient budding yeast with an Arabidopsis cDNA library. CycJ18 shares only 20% identity in its conserved cyclin box domain with other cyclins, and is predominantly expressed in young seedlings. CycJ18 is a member of a potential new plant cyclin class. PMID- 11389962 TI - SV40 Immortalization of feline fibroblasts as targets for MHC-restricted cytotoxic T-cell assays. AB - CTL assays in outbred cats have been difficult to perform because of a lack of a good source of syngeneic target cell. Primary fibroblasts from cats are widely used as target cells for MHC-restricted cytotoxic T-cell (CTL) assays, but their limited life-spans of 8-10 culture passages can be problematic for longitudinal studies. To circumvent the life-span limitations of primary fibroblast cultures, we developed a procedure for immortalizing feline primary fibroblast cells by transfection with a molecular clone of simian virus 40 (SV40). Fibroblast cultures from skin biopsies of 28 cats were immortalized using this procedure and have been passaged for longer than 6 months without showing any phenotypic difference from the original primary cells. Non-SV40 transfected feline fibroblasts from a selection of animals in the same group survived for only 6-8 weeks before reaching senescence. The immortalized fibroblasts expressed SV40 T antigen and Class I MHC protein, and were successfully used as target cells in 51Cr release CTL assays in feline immunodeficiency virus (FIV)-infected cats and in vitro stimulated allogeneic T-cell cultures. PMID- 11389964 TI - Modulation of the host immune system by phosphorylcholine-containing glycoproteins secreted by parasitic filarial nematodes. AB - Phosphorylcholine (PC) is increasingly becoming recognised as a carbohydrate associated component of a wide variety of procaryotic and eucaryotic pathogens. Studies employing nematode PC-containing molecules indicate that it possesses a plethora of immunomodulatory activities. ES-62 is a PC-containing glycoprotein, which is secreted by the rodent filarial nematode Acanthocheilonema viteae and which provides a model system for the dissection of the mechanisms of immune evasion induced by related PC-containing glycoproteins expressed by human filarial nematodes. At concentrations equivalent to those found for PC-containing molecules in the bloodstream of parasitised humans, ES-62 is able to inhibit antigen receptor-stimulated proliferation of B and T lymphocytes in vitro and in vivo. The active component of ES-62 appears to be PC, as PC conjugated to albumin or even PC alone broadly mimic the results obtained with ES-62. PC-induced impaired lymphocyte responsiveness appears to reflect uncoupling of the antigen receptors from key intracellular proliferative signalling events such as the phosphoinositide 3-kinase, protein kinase C and Ras mitogen-activating protein kinase pathways. Although PC-ES-62 can desensitise B and T cells, not all cells are affected, and in fact it is still possible to generate an antibody response to the molecule. Dissection of this response indicates that it is of the TH-2 type. This appears to reflect the ability of ES-62 to direct the polarity of the T cell response by suppressing the production of proinflammatory cytokines, inducing the induction of anti-inflammatory cytokines and by driving the maturation of dendritic cells that direct TH-2 T cell responses. PMID- 11389965 TI - Expression of cGMP-binding cGMP-specific phosphodiesterase (PDE5) in mouse tissues and cell lines using an antibody against the enzyme amino-terminal domain. AB - We have produced a polyclonal antibody that specifically recognizes cGMP-binding cGMP-specific phosphodiesterase (PDE5). The antibody was raised in rabbit using as immunogen a fusion protein, in which glutathione S-transferase was coupled to a 171 amino acid polypeptide of the N-terminal region of bovine PDE5. The antibody is able to immunoprecipitate PDE5 activity from mouse tissues and neuroblastoma extracts while it has no effect on all other PDE isoforms present in the extracts. PDE5 activity recovered in the immunoprecipitates retains its sensitivity to specific inhibitors such as zaprinast (IC(50)=0.6 microM) and sildenafil (IC(50)=3.5 nM). Bands of the expected molecular mass were revealed when solubilized immunoprecipitates were analysed in Western blots. The antibody selectively stained cerebellar Purkinje neurones, which are known to express high levels of PDE5 mRNA. Western blot analysis of mouse tissues revealed the highest expression signal in mouse lung, followed by heart and cerebellum, while a lower signal was evident in brain, kidney and a very low signal was present in the liver. In the hybrid neuroblastoma-glioma NG108-15 cells the antibody revealed a high PDE5 induction after dibutyryl-cAMP treatment. PMID- 11389966 TI - Multipathways for transdifferentiation of human prostate cancer cells into neuroendocrine-like phenotype. AB - The neuroendocrine (NE) cell is a minor cell population in normal human prostate glands. The number of NE cells is increased in advanced hormone-refractory prostate carcinomas (PCA). The mechanism of increased NE cell population in these advanced tumors is poorly understood. We examined molecular mechanisms which may be involved in the regulation of the transdifferentiation process of human PCA cells leading to a NE phenotype. We compared PCA cell lines LNCaP and PC-3 in the following medium conditions: steroid-reduced (SR), interleukin-6 (IL-6) supplemented, or dibutyrate cAMP (db-cAMP)-supplemented. We found that androgen responsive C-33 LNCaP cells responded to all treatments, having a neuronal-like morphology. In contrast, C-81 LNCaP cells, having a decreased androgen responsiveness, had a less pronounced effect although followed a similar trend. Androgen-unresponsive PC-3 cells showed little change in their morphology. Grown in the SR condition, the level of neuron-specific enolase (NSE), a marker of neuronal cells, was upregulated in C-33 LNCaP cells, while to a lesser degree in the presence of IL-6. In the presence of db-cAMP, the NSE level in C-33 cells was decreased, lower than that in control cells. An opposite effect was observed for C-81 LNCaP cells. Nevertheless, the NSE level was only elevated in db-cAMP treated PC-3 cells, but no change was found in PC-3 cells grown in the SR- or IL 6-supplemented medium. Thus, a similar gross phenotypic change may correlate with differential molecular expressions. We also analyzed the expression of protein tyrosine phosphatase alpha (RPTPalpha) since it plays a critical role in normal neuronal differentiation and signaling. Our results showed that the expression of RPTPalpha correlates with the NE phenotypic change of LNCaP cells in the SR condition. In summary, our data clearly show that the molecular process by which cultured human prostate cancer cells undergo a transdifferentiation process to a NE cell-like phenotype is accompanied by differential expressions of different markers, and a gross NE cell-like phenotype can occur by exposing PCA cells to different pharmacological agents. PMID- 11389968 TI - Micro-Raman characterisation of the R to T state transition of haemoglobin within a single living erythrocyte. AB - We present the first recorded Raman spectra of haemoglobin in both the R and T states from within a single living erythrocyte using 632.8 nm excitation. Bands characteristic of low spin haems are observed in oxygenated and carboxylated erythrocytes at approx. 1636 (nu(10)), 1562-1565 (nu(2)), 1250-1245 cm(-1) (nu(13)) and 1226-1224 cm(-1) (nu(5)+nu(8)). The spectra of deoxygenated and methaemoglobin erythrocytes have characteristic high spin bands at approx. 1610 1606 cm(-1) (nu(10)), 1582-1580 (nu(37)), 1547-1544 (nu(11)), 1230-1220 cm(-1) (nu(13)) and 1215-1210 cm(-1) (nu(5)+nu(8)). Bands at 1172 (nu(30)), 976 (nu(45)) and 672 (nu(7)) cm(-1) appear to be enhanced at 632.8 nm in low spin haems. The oxidation state marker band (nu(4)) at 1364-1366 cm(-1) appeared invariant within this domain in all single cells and conditions investigated contrary to other resonance Raman studies on haem isolates. The information gained by in vivo single erythrocyte molecular analysis has important ramifications to the understanding of fundamental physiological processes and may have applications in the diagnosis and treatment of red blood cell disorders. PMID- 11389967 TI - 3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1) induces caspase-dependent apoptosis in mononuclear cells. AB - 3-Amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), one of the tryptophan pyrolysates, is a dietary carcinogen and is formed in cooked meat and fish in our daily diet. Trp-P-1 will affect the cells in the blood circulation system before it causes carcinogenicity in target organs such as the liver. In this study, the cytotoxicity of Trp-P-1 was investigated in mononuclear cells (MNCs) from blood. Trp-P-1 (10-15 microM) decreased cell viability and induced apoptosis characterized both by morphological changes and by DNA fragmentation 4 h after treatment. DNA fragmentation was also observed following treatment at 1 nM after 24 h in culture. This result suggested that apoptosis would occur in the body following unexpected intake of foods containing Trp-P-1. To determine the mechanism of apoptosis, we investigated the activation of the caspase cascade in MNCs. Trp-P-1 (10-15 microM) activated the caspase cascade, i.e. the activity of caspase-3, -6, -7, -8 and -9 increased dose-dependently using peptide substrates, the active forms of caspase-3, -8 and -9 were detected by immunoblotting, and cleavage of poly(ADP-ribose) polymerase and protein kinase C-delta as the intracellular substrates for caspases was observed. A peptide inhibitor of caspase-8 completely suppressed activation of all other caspases, while an inhibitor of caspase-9 did not. These results indicated that caspase-8 may act as an apical caspase in the Trp-P-1-activated cascade. PMID- 11389969 TI - Specific secretion of gel-forming mucins and TFF peptides in HT-29 cells of mucin secreting phenotype. AB - Trefoil factor family (TFF) peptides are typical secretory products of mucin producing cells, e.g. of the gastrointestinal tract. Here, the expression and secretion of mucins and TFF peptides was studied in the HT-29 cell line throughout cellular growth and differentiation in relation to a mucin-secreting (HT-29 MTX) or an enterocyte-like (HT-29 G(-)) phenotype. mRNAs of several MUC and TFF genes were expressed in both cell subpopulations. However, for most MUC and TFF genes, the expression appeared strongly induced with the differentiation into the mucin-secreting phenotype. On the other hand, TFF2 was specifically expressed in the mucin-secreting HT-29 MTX cells. The differentiation of HT-29 MTX cells into the mucin-secreting phenotype was characterised by secretion of the gel-forming mucins MUC2, MUC5AC, and MUC5B, however, according to a different pattern in the course of differentiation. A significant amount of TFF1 and TFF3 was secreted after differentiation, also according to a different pattern, whereas TFF2 was only faintly detected. Secretagogues, known to induce the secretion of mucus, increased the secretion of all three TFF peptides. In contrast, neither a secretory mucin nor a TFF peptide was found in the culture medium of HT-29 G(-) cells. Overlay assays indicated that HT-29 MTX mucins bound to secretory peptides of HT-29 MTX cells with relative molecular mass similar to TFF peptides. TFF1 and TFF3 were specifically localised in the mucus layer of HT 29 MTX cells by confocal microscopy. Finally, the secretion of TFF peptides and mucins appears as a co-ordinated process which only occurs after differentiation into goblet cell-like phenotype. PMID- 11389970 TI - Effect of the beta(3)-adrenergic agonist Cl316,243 on functional differentiation of white and brown adipocytes in primary cell culture. AB - We investigated the effect of the specific beta(3)-adrenergic receptor agonist CL 316,243 (CL) on proliferation and functional differentiation of the Siberian hamster (Phodopus sungorus) white and brown preadipocytes in primary cell culture. Proliferation of both white and brown preadipocytes was stimulated by a general beta-adrenergic agonist (isoproterenol) but not by CL. Lipolysis of differentiated white and brown adipocytes was stimulated similarly by CL with maximum effect at 10 nM. Thermogenic properties of cells were assessed by immunodetection of UCP-1, the brown adipocyte specific uncoupling protein, and measurement of cytochrome c oxidase (COx) activity as an index of mitochondrial capacity. UCP-1 content was largely increased by CL in BAT but not in WAT cultures. Basal UCP-2 mRNA levels were similar in WAT and BAT cultures and increased by both CL and isoproterenol. COx activity of BAT cultures was twice as high as that of WAT cultures but in neither cell culture system could it be increased by beta-adrenergic stimulation. We suggest (i) that white and brown preadipocyte proliferation is increased in vitro via beta1 or beta(2), but not beta(3)-adrenergic pathways, (ii) that white and brown preadipocytes represent different cell types, and (iii) that in vitro beta-adrenergic stimulation it is not sufficient to induce complete thermogenic adaptation of brown adipocytes. PMID- 11389971 TI - Confocal microscopic study of GABA(A) receptors in Xenopus oocytes after rat brain mRNA injection: modulation by tyrosine kinase activity. AB - The expression of GABA(A) receptors in Xenopus oocytes injected with rat brain mRNA was studied by immunocytochemistry and evaluation of the distribution of fluorescent probes at the confocal microscope. The beta(2/3) subunit distributed exclusively on the membrane at the animal pole of the oocytes. Treatment of oocytes for 20 min with the protein tyrosine kinase inhibitor genistein, 200 microM, resulted in a lower presence of GABA(A) receptors on the membrane. The inactive genistein analogue daidzein, 200 microM, had no effect even with a 30 min treatment. Alkaline phosphatase but not a protein tyrosine phosphatase, when injected into oocytes, reduced GABA(A) receptor membrane expression. The data indicate that protein tyrosine phosphorylation modulates the expression on the plasma membrane of presynthesized GABA(A) receptors. PMID- 11389972 TI - Nitric oxide regulates actin reorganization through cGMP and Ca(2+)/calmodulin in RAW 264.7 cells. AB - Nitric oxide (NO) has been reported to be involved in the regulation of pseudopodia formation, phagocytosis and adhesion in macrophages through the reorganization of actin. In the present study, we directly separated the globular (G) and filamentous (F) actin from quiescent or NO-stimulated macrophage-like cell line RAW 264.7 cells in order to investigate the dynamic redistribution of actin pools. We also focused on the regulatory mechanisms of actin assembly, induced by NO and its possible subsequent signaling pathway. We showed that predominant G-actin coexisted with Triton X-100-insoluble filamentous (TIF) and Triton X-100-soluble filamentous actin in resting RAW 264.7 cells. The exogenous NO produced by (+/-)-(E)-2-[(E)-hydroxyimino]-6-methoxy-4-methyl-5-nitro-3 hexenamide (NOR1), the endogenous NO induced by lipopolysaccharide (LPS) plus interferon-gamma (IFNgamma), and dibutyryl-cGMP increased the contents of TIF actin in dose- and time-dependent manners and altered its morphology. The increase in the TIF-actin contents induced by NOR1 or LPS plus IFNgamma was efficiently blocked by the radical scavenger 2-(4-carboxyphenyl)-4,4,5,5 tetramethylimidazoline-1-oxyl 3-oxide and the soluble guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one or the arginine analogue N(G) monomethyl-L-arginine acetate, respectively. Preincubation with the calmodulin antagonist W-7 almost completely blocked the NO-induced TIF-actin increase and morphological change. On the other hand, preincubation with C3 transferase, an inhibitor of Rho protein, efficiently prevented the change in cell morphology, but had no effect on the TIF-actin increase. We postulate that cGMP and subsequent Ca(2+)/calmodulin may be key regulators of actin reorganization in NO stimulated RAW 264.7 cells. PMID- 11389973 TI - Possible mechanisms regulating ATP- and thimerosal-induced Ca(2+) oscillations in the HSY salivary duct cell line. AB - The ATP-induced oscillatory changes in cytosolic Ca(2+) concentration ([Ca(2+)](i)) were analysed in HSY cells, a salivary ductal cell line from human parotid, using a fluorescence ratio imaging system. At concentrations higher than 1 microM, ATP caused sinusoidal [Ca(2+)](i) oscillations due to the periodic release and reuptake of Ca(2+) by intracellular Ca(2+) stores. The phorbol ester 4beta-phorbol 12,13-dibutyrate (PDBu) changed the [Ca(2+)](i) oscillations to a single spike. The inhibitory effect of PDBu on the [Ca(2+)](i) signals was reversed by protein kinase C (PKC) inhibitors such as staurosporine and chelerythrine chloride. However, preincubation of the cells with the PKC inhibitors did not affect the pattern of the ATP-induced [Ca(2+)](i) oscillations. The desensitization of the [Ca(2+)](i) response observed during prolonged stimulation with ATP was also not prevented by the PKC inhibitors. Incubation of HSY cells with the sulphydryl reagent thimerosal, which enhances the sensitivity of inositol 1,4,5-trisphosphate (IP(3)) receptors, caused repetitive Ca(2+) release from intracellular Ca(2+) stores resulting in baseline spikes of [Ca(2+)](i). The thimerosal-induced [Ca(2+)](i) oscillations did not change in the presence of PDBu and the phospholipase C inhibitor U73122. Thus, we could not provide evidence that negative feedback by PKC plays a central role in the regulation of ATP-induced [Ca(2+)](i) oscillations. These results suggest that the [Ca(2+)](i) oscillations, at least the baseline spikes, in HSY cells can be generated without stimulating the formation of IP(3). PMID- 11389974 TI - Vitamin D(3) enhances the expression of I-mfa, an inhibitor of the MyoD family, in osteoblasts. AB - I-mfa (inhibitor of the MyoD family) is a transcription modulator that binds to and suppresses the transcriptional activity of MyoD family members. I-mfa transcripts are expressed in sclerotome, suggesting a role of I-mfa in skeletogenesis. The aim of this study was to examine the expression and regulation of I-mfa in osteoblasts. We found that I-mfa is expressed at a low level in an osteoblast-like cell line, MC3T3E1, and a pluripotent differentiation modulator, 1,25-dihydroxyvitamin D(3), specifically enhanced I-mfa mRNA expression. This effect was completely blocked by the presence of an RNA polymerase inhibitor, but not by a protein synthesis inhibitor, suggesting that 1,25-dihydroxyvitamin D(3) upregulates transcription of the I-mfa gene without requirement for new protein synthesis. Western blot analysis indicated that 1,25 dihydroxyvitamin D(3) increased the I-mfa protein levels severalfold in MC3T3E1 cells. I-mfa expression was also observed in primary mouse calvaria cells and ROS17/2.8 cells and 1,25-dihydroxyvitamin D(3) enhanced I-mfa expression in these cells. These data indicate that I-mfa is a novel transcriptional regulator gene expressed in osteoblasts and that its level is under the control of 1,25 dihydroxyvitamin D(3). PMID- 11389975 TI - Glycosylation of human CRLR at Asn123 is required for ligand binding and signaling. AB - Calcitonin receptor-like receptor (CRLR) constitutes either a CGRP receptor when complexed with receptor activity-modifying protein 1 (RAMP1) or an adrenomedullin receptor when complexed with RAMP2 or RAMP3. RAMP proteins modify the glycosylation status of CRLR and determine their receptor specificity; when treated with tunicamycin, a glycosylation inhibitor, CHO-K1 cells constitutively expressing both RAMP2 and CRLR lost the capacity to bind adrenomedullin. Similarly, in HEK293 EBNA cells constitutively expressing RAMP1/CRLR receptor complex CGRP binding was remarkably inhibited. Whichever RAMP protein was co expressing with CRLR, the ligand binding was sensitive to tunicamycin. There are three putative Asn-linked glycosylation sites in the extracellular, amino terminal domain of CRLR at positions 66, 118 and 123. Analysis of CRLR mutants in which Gln was substituted for selected Asn residues showed that glycosylation of Asn123 is required for both the binding of adrenomedullin and the transduction of its signal. Substituting Asn66 or Asn118 had no effect. FACS analysis of cells expressing FLAG-tagged CRLRs showed that disrupting Asn-linked glycosylation severely affected the transport of the CRLR protein to the cell surface on N66/118/123Q mutant, and slightly reduced the level of the cell surface expression of N123Q mutant compared with wild-type CRLR. But other single mutants (N66Q, N118Q) had no effect for other single mutants. Our data shows that glycosylation of Asn66 and Asn118 is not essential for ligand binding, signal transduction and cell surface expression, and Asn123 is important for ligand binding and signal transduction rather than cell surface expression. It thus appears that glycosylation of Asn123 is required for CRLR to assume the appropriate conformation on the cell surface through its interaction with RAMPs. PMID- 11389976 TI - Lipopolysaccharide-induced cell cycle arrest in macrophages occurs independently of nitric oxide synthase II induction. AB - Lipopolysaccharide (LPS, a Gram-negative bacterium cell wall component) is a potent macrophage activator that inhibits macrophage proliferation and stimulates production of nitric oxide (NO) via NO synthase II (NOSII). We investigated whether NO mediates the LPS-stimulated cell cycle arrest in mouse bone marrow derived macrophages (BMM). The addition of the NO donor DETA NONOate (200 microM) inhibited BMM proliferation by approx. 80%. However, despite NO being an antimitogen, LPS was as potent at inhibiting proliferation in BMM derived from NOSII-/- mice as from wild-type mice. Consistent with these findings, LPS-induced cell cycle arrest in normal BMM was not reversed by the addition of the NOSII inhibitor S-methylisothiourea. Moreover, in both normal and NOSII-/- BMM, LPS inhibited the expression of cyclin D1, a protein that is essential for proliferation in many cell types. Despite inhibiting proliferation DETA NONOate had no effect on cyclin D1 expression. Our data indicate that while both LPS and NO inhibit BMM proliferation, LPS inhibition of BMM proliferation can occur independently of NOSII induction. PMID- 11389977 TI - Thromboxane A(2) receptor mediated activation of the mitogen activated protein kinase cascades in human uterine smooth muscle cells. AB - Both thromboxane (TX) A(2) and 8-epi prostaglandin (PG) F(2alpha) have been reported to stimulate mitogenesis of vascular smooth muscle (SM) in a number of species. However, TXA(2) and 8-epiPGF(2alpha) mediated mitogenic signalling has not been studied in detail in human vascular SM. Thus, using the human uterine ULTR cell line as a model, we investigated TXA(2) receptor (TP) mediated mitogenic signalling in cultured human vascular SMCs. Both the TP agonist U46619 and 8-epiPGF(2alpha) elicited time and concentration dependent activation of the extracellular signal regulated kinase (ERK)s and c-Jun N-terminal kinase (JNK)s in ULTR cells. Whereas the TP antagonist SQ29548 abolished U46619 mediated signalling, it only partially inhibited 8-epiPGF(2alpha) mediated ERK and JNK activation in ULTR cells. Both U46619 and 8-epiPGF(2alpha) induced ERK activations were inhibited by the protein kinase (PK) C, PKA and phosphoinositide 3-kinase inhibitors GF109203X, H-89 and wortmannin, respectively, but were unaffected by pertussis toxin. In addition, U46619 mediated ERK activation in ULTR cells involves transactivation of the epidermal growth factor (EGF) receptor. In humans, TXA(2) signals through two distinct TP isoforms. In investigating the involvement of the TP isoforms in mitogenic signalling, both TPalpha and TPbeta independently directed U46619 and 8-epiPGF(2alpha) mediated ERK and JNK activation in human embryonic kidney (HEK) 293 cells over-expressing the individual TP isoforms. However, in contrast to that which occurred in ULTR cells, SQ29548 abolished 8-epiPGF(2alpha) mediated ERK and JNK activation through both TPalpha and TPbeta in HEK 293 cells providing further evidence that 8 epiPGF(2alpha) may signal through alternative receptors, in addition to the TPs, in human uterine ULTR cells. PMID- 11389979 TI - Identification and characterisation of PEX6 orthologues from plants. AB - The sunflower (Helianthus annuus) orthologue of PEX6, an AAA ATPase essential for the biogenesis of peroxisomes in yeasts and mammals, was isolated. HaPex6p is immunologically related to Pichia pastoris Pex6p. Like other genes involved in peroxisome biogenesis and function HaPEX6 mRNA and protein levels peak in early post-germinative growth and mRNA levels also increase in senescent tissue. HaPEX6 identifies probable orthologues in Arabidopsis and rice. PMID- 11389978 TI - Association of protein kinase C(lambda) with adducin in 3T3-L1 adipocytes. AB - There is evidence that the atypical protein kinases C (PKC(lambda), PKC(zeta)) participate in signaling from the insulin receptor to cause the translocation of glucose transporters from an intracellular location to the plasma membrane in adipocytes. In order to search for downstream effectors of these PKCs, we identified the proteins that were immunoprecipitated by an antibody against PKC(lambda/zeta) from lysates of 3T3-L1 adipocytes through peptide sequencing by mass spectrometry. The data show that PKC(lambda) is the major atypical PKC in these cells. Moreover, an oligomeric complex consisting of alpha- and gamma adducin, which are cytoskeletal proteins, coimmunoprecipitated with PKC(lambda). Association of the adducins with PKC(lambda) was further indicated by the finding that the adducins coimmunoprecipitated proportionally with PKC(lambda) in repeated rounds of immunoprecipitation. Such an association is consistent with literature reports that the adducins contain a single major site for PKC phosphorylation in their carboxy termini. Using antibody against the phospho form of this site for immunoblotting, we found that insulin caused little or no increase in the phosphorylation of this site on the adducins in a whole cell lysate or on the small portion of the adducins that coimmunoprecipitated with PKC(lambda). PKC(lambda) and the adducins were located in both the cytosol and subcellular membranous fractions. The binding of PKC(lambda) to adducin may function to localize PKC(lambda) in 3T3-L1 adipocytes. PMID- 11389980 TI - Modulated release of cyclosporine from soluble vinyl pyrrolidone--hydroxyethyl methacrylate copolymer hydrogels. A correlation of 'in vitro' and 'in vivo' experiments. AB - Soluble, uncrosslinked and high molecular weight copolymers of vinylpyrrolidone, VP, with 2-hydroxyethyl methacrylate, HEMA, prepared by free radical copolymerization, are proposed as supports for the modulated release of the immunosuppressor cyclosporine. Two copolymeric systems with copolymer compositions f(VP)=0.52 (namely VP--HEMA 60--40) and 0.42 (VP--HEMA 40--60) have been prepared and tested in vitro and in vivo using rats as animal model. Micellar electrokinetic capillary chromatography, MEKC, has been used for the simultaneous detection of the polymer reabsorption and the drug release for the in vitro experiments. The composition and microstructural distribution of the copolymer system controls the solubilization rate which modulates the in vitro release of the drug (with time profiles from a few days to several weeks for the VP--HEMA 60--40 and 40--60, respectively) and the in vivo response that correlates with the previous in vitro results: the more hydrophobic implant (VP- HEMA 40--60) reverts the immune response more slowly (2--4 weeks) compared to the more hydrophilic one (VP--HEMA 60--40, 1--2 weeks). PMID- 11389981 TI - Development of poly-(D,L-lactide--coglycolide) microsphere formulations containing recombinant human vascular endothelial growth factor to promote local angiogenesis. AB - Although preclinical animal studies have demonstrated the utility of recombinant human vascular endothelial growth factor (rhVEGF) in promoting neovascularization in regions of ischemia, rhVEGF systemic administration did not provide clinical benefit to patients in recent placebo-controlled Phase II clinical trials. The amount of rhVEGF localized in the ischemic region after systemic administration is minimal and does not persist for more than 1 day. A greater persistence of rhVEGF at the region of ischemia may provide an increased angiogenesis with the eventual formation of patent blood vessels to restore nourishment to the tissues. We sought to develop a formulation of rhVEGF in poly(D,L-lactide--co-glycolide) (PLG) microspheres that would provide a continuous local delivery of intact protein. A stable formulation of rhVEGF for encapsulation contained a small amount of a stabilizing sugar, trehalose. Addition of excess trehalose increased the rate of release from the PLG. In addition, PLG with free acid end groups appeared to retard the initial release of rhVEGF by associating with it through ionic interactions at the positively charged heparin binding domain. rhVEGF was released continuously for 21 days with a very low (less than 10%) initial burst. The released rhVEGF aggregated and hydrolyzed over time and lost heparin affinity but not receptor affinity. The compression molding of rhVEGF PLG microspheres into disks yielded formulations with a low initial release and a lag of 10 days followed by complete release. The PLG microsphere formulations were assessed in the corneal implant model of angiogenesis and generated a dose-dependent angiogenic response. These formulations were also administered intravitreally and subretinally, generating local neovascularization comparable to the human disease states, vitroretinopathy and age-related macular degeneration, respectively. The rhVEGF PLG formulations may increase local angiogenesis without systemic side effects and may also be useful in the development of ocular disease models. PMID- 11389982 TI - Surface modifications and molecular imprinting of polymers in medical and pharmaceutical applications. AB - Recent developments in the field of biomaterials are based on molecular design of polymers with improved surface and bulk properties. Novel techniques of surface modification by addition of tethered chains can lead to materials with the ability to recognize biological and pharmaceutical compounds. Methods based on molecular imprinting can increase the recognition capabilities of such systems. Chain tethering can also can improve the mucoadhesive behavior of a delivery device and the effectiveness of a drug by allowing targeting and localization of a drug at a specific site. Acrylic-based hydrogels are well-suited for mucoadhesion due to their flexibility and nonabrasive characteristics which reduce damage-causing attrition to the tissues in contact. However, the adhesive and drug delivery capabilities of these devices can continue to be improved as presently known bioadhesive materials are modified and more bioadhesive materials are discovered. Tethering of long PEG chains on PAA hydrogels and their copolymers can be achieved by grafting reactions involving thionyl chloride, followed by PEG grafting. The ensuing materials exhibit mucoadhesive properties due to enhanced anchoring of the chains with the mucosa. Theoretical calculations can lead to optimization of the tethered structure. PMID- 11389983 TI - Release of PEGylated granulocyte-macrophage colony-stimulating factor from chitosan/glycerol films. AB - We have prepared a new formulation for mucosal delivery of GM-CSF or PEGylated GM CSF based on a chitosan carrier plus added glycerol to control the rate of release of the protein. Thin dry films comprised of various weight ratios of chitosan to glycerol and containing either granulocyte-macrophage colony stimulating factor (GM-CSF) or PEGylated GM-CSF, PEG-(GM-CSF), were prepared. The amount of GM-CSF or PEG-(GM-CSF) released from the chitosan/glycerol films was determined using size exclusion high performance liquid chromatography (HPLC SEC). The amount of PEG-(GM-CSF) released from the films decreased with an increase in the amount of glycerol present in the film. In parallel with this, films with higher glycerol content exhibited a lower degree of equilibrium swelling when immersed in release media. pH measurements of the release media and analysis of the dried films by Fourier-transform infrared spectroscopy (FTIR) suggested that the amount of residual acetic acid in the dry films decreased as the glycerol content increased. This indicates that glycerol may act by displacing and releasing bound acetic acid from the chitosan molecules, resulting in chitosan--glycerol hydrogen bond formation as the film dries. Further, it was found that the release rate and the amount of PEG-(GM-CSF) released decreased with increasing molecular weight of the conjugated PEG. This effect was not observed with films containing physical mixtures of PEG and GM-CSF. The decrease in the fraction of PEG-(GM-CSF) released with increasing PEG molecular weight is believed to be due to the increased steric hindrance of the PEGylated protein molecule during its diffusion out of the swollen chitosan/glycerol film. PMID- 11389984 TI - Biodegradable hydrogels based on stereocomplex formation between lactic acid oligomers grafted to dextran. AB - A novel hydrogel system in which crosslinking is established by stereocomplex formation between lactic acid oligomers of opposite chirality is proposed. To investigate the feasibility of this novel system, we first investigate whether there is an operation window where lactic acid oligomers in either the D- or L form do not give a crystalline phase, whereas in a blend of the D- and L-form stereocomplex formation occurs. Therefore, D- and L-lactic acid oligomers with different degrees of polymerization (DP) were prepared and analyzed using DSC. It was shown that crystallinity was present in D- or L-oligomers with DP > or = 11. On the other hand, in blends of D- and L-oligomers of lactic acid crystallinity (stereocomplexation) was already observed at a DP > or = 7. In the next step, L- and D-lactic acid oligomers were coupled via their terminal hydroxyl group to dextran, yielding dex-(L)lactate and dex-(D)lactate, respectively. Upon dissolving each product in water separately and mixing the solutions, a hydrogel is formed at room temperature as demonstrated by rheological measurements. The storage modulus of the obtained hydrogel strongly decreased upon heating to 80 degrees C, while it was restored upon cooling to 20 degrees C demonstrating the thermo-reversibility and the physical nature of the cross-links. The storage modulus of the gels depends on the degree of polymerization of the lactate acid grafts and their degree of substitution on dextran. Interestingly, gel formation was favored when one lactic oligomer was coupled via its hydroxyl group whereas the oligomer of opposite chirality was coupled via its carboxylic acid group. This is ascribed to the parallel packing of the oligomers in stereocomplexes. PMID- 11389985 TI - The coiled coils in the design of protein-based constructs: hybrid hydrogels and epitope displays. AB - Recombinant DNA technology provides a powerful tool for producing protein-based biomaterials. Genetically engineered coiled coils have been used as a structural module for the construction of a variety of bio-based systems useful in drug delivery studies. Two of such approaches developed in the authors' laboratory were described here. One approach was to assemble hybrid hydrogels from coiled coil protein domains and synthetic polymers. Preliminary results showed that temperature-sensitive volume transition of the hybrid hydrogels could be triggered by the thermal unfolding of the engineered coiled coil protein domains. The other approach, discussed in detail, was to construct an epitope display model system based on a coiled coil stem loop peptide self-assembled on a solid substrate. This model construct displayed a constrained nonapeptide sequence, which was found to mediate specific binding with immunocompetent cells bearing complementary surface receptors. These novel approaches will likely find important applications in the rational design of more effective drug delivery systems. PMID- 11389986 TI - N-isopropylacrylamide copolymers for the preparation of pH-sensitive liposomes and polymeric micelles. AB - Hydrophobically-modified copolymers of N-isopropylacrylamide bearing a pH sensitive moiety were investigated for the preparation of pH-responsive liposomes and polymeric micelles. The copolymers having the hydrophobic anchor randomly distributed within the polymeric chain were found to more efficiently destabilize egg phosphatidylcholine (EPC)/cholesterol liposomes than the alkyl terminated polymers. Release of both a highly-water soluble fluorescent contents marker, pyranine, and an amphipathic cytotoxic anti-cancer drug, doxorubicin, from copolymer-modified liposomes was shown to be dependent on pH, the concentration of copolymer, the presence of other polymers such as polyethylene glycol, and the method of preparation. Both polymers were able to partially stabilize EPC liposomes in human serum. These polymers were found to self-assemble to form micelles. The critical association concentration was low (9--34 mg/l) and influenced by the position of the alkyl chains. In phosphate buffered saline, the micelles had a bimodal size distribution with the predominant population having a mean diameter of 35 nm. The polymeric micelles were studied as a delivery system for the photosensitizer aluminum chloride phthalocyanine, (AlClPc), currently evaluated in photodynamic therapy. pH-Responsive polymeric micelles loaded with AlClPc were found to exhibit increased cytotoxicity against EMT-6 mouse mammary cells in vitro than the control Cremophor EL formulation. PMID- 11389987 TI - Pronounced activity of enzymes through the incorporation into the core of polyion complex micelles made from charged block copolymers. AB - Compartmentalization of enzymes in the nanometric-scaled container, to improve their stability and availability, has recently attracted a strong interest in the field of pharmaceutics. In this study, the enzymatic activity of lysozyme in the core of polyion complex (PIC) micelles, which were formed from egg white lysozyme and poly(ethylene glycol)-poly(alpha,beta-aspartic acid) block copolymer (PEG P(Asp)), was evaluated using a colorimetric method. Apparent enzymatic activity of lysozyme entrapped in the core of PIC micelles remarkably increased compared to that of free lysozyme, which is mainly attributed to a decrease in the observed Michaelis constant (K(m,obs)). The reciprocal of the K(m,obs) values nicely correlated to the corona thickness of PIC micelles, suggesting that the corona layer of PIC micelle may act as the reservoir of the substrate, p nitrophenyl penta-N-acetyl-beta-chitopentaoside. This result indicates that the enzymatic activity can be controlled by changing the corona thickness of PIC micelles through a variation in the mixing ratio of PEG-P(Asp) to lysozyme. This type of PIC micelle system entrapping enzyme in the core might be useful for the design of diagnostic as well as targetable therapeutic systems of enzyme including antibody-directed enzyme prodrug therapy (ADEPT). PMID- 11389988 TI - Tissue engineering as a platform for controlled release of therapeutic agents: implantation of microencapsulated dopamine producing cells in the brains of rats. AB - Tissue engineering can lead to novel controlled release devices and controlled release strategies (e.g., of growth factors) can enhance the performance of tissue engineered constructs. There are however a number of technical challenges that must be overcome before these goals can be realized. The apparently 'simple' challenge of implanting the device (e.g., capsules) in the optimal site must be met. In addition, adequate nutrient supply to the capsules is required to maintain cell viability. To illustrate this problem we describe a guide and delivery cannula technique to provide reliable and reproducible delivery of up to 120 PC12 cell containing capsules into the caudate putamen (CPu). Microencapsulation of mammalian cells is potentially a powerful means of delivering therapeutically important molecules such as insulin. It can also have numerous applications as a platform for gene therapy. However, realizing this potential has been more difficult than first anticipated. PMID- 11389989 TI - Covalently conjugated VEGF--fibrin matrices for endothelialization. AB - Vascular endothelial growth factor (VEGF) is a key factor in endothelial cell biology and blood vessel formation and a candidate therapeutic for the stimulation of angiogenesis-dependent tissue regeneration. The objective of this study was to confer the angiogenic activity of VEGF(121) upon the biomaterial fibrin, a natural substrate for endothelial cell growth and clinically accepted as 'fibrin glue'. To achieve this, we engineered fibrin-based hydrogels that were covalently modified with VEGF(121). Our laboratory has recently developed novel methodology that allows the covalent incorporation of exogenous bioactive peptides by the transglutaminase activity of factor XIIIa into fibrin during coagulation. Here, this ability of factor XIIIa to crosslink additional proteins within fibrin was employed to covalently incorporate VEGF(121). By recombinant DNA methodology, a mutant VEGF(121) variant, alpha(2)-PI(1--8)-VEGF(121), which contains an additional factor XIIIa substrate sequence NQEQVSPL at the aminoterminus, was expressed in E. coli. In soluble form, the mutant protein fully retained its mitogenic activity for endothelial cells. Using (125)I-labeled alpha(2)-PI(1--8)-VEGF(121), its covalent incorporation and the efficiency of incorporation into fibrin was demonstrated and characterized. The immobilized, fibrin-conjugated VEGF(121) protein remained an active and very efficient mitogen for human endothelial cells grown on two-dimensional VEGF(121)-modified fibrin surfaces, and the incorporation of increasing amounts of alpha(2)-PI(1--8) VEGF(121) resulted in dose-dependent enhancement of endothelial cell growth. The VEGF-modified fibrin matrices can be formed as injectable gels in a single-step reaction under physiological conditions in vivo. When used as a ingrowth matrix, such VEGF incorporating materials could be useful in a variety of clinical situations that require an angiogenic response into an ischemic region or inplant. PMID- 11389990 TI - Recent progress in gene delivery using non-viral transfer complexes. AB - The delivery of genetic material into cells is a field that is expanding very rapidly. Non-viral delivery methods, especially ones that focus on the use of chemical agents complexed with genetic material, are the focus of this mini review. More-recent uses of known transfection agents such as poly(ethylenimine), poly(L-lysine), and various liposomes are discussed, and some novel approaches (both chemical and methodical) are reviewed as well. A very brief look at how non viral gene delivery research is being aimed at the clinic is also included. PMID- 11389991 TI - Regulatory perspectives on in vitro (dissolution)/in vivo (bioavailability) correlations. AB - In vitro dissolution has been extensively used as a quality control tool for solid oral dosage forms. In several cases, however, it is not known whether one can predict the in vivo performance of these products from in vitro dissolution data. In an effort to minimize unnecessary human testing, investigations of in vitro/in vivo correlations (IVIVC) between in vitro dissolution and in vivo bioavailability are increasingly becoming an integral part of extended release (ER) drug product development. This increased activity in developing IVIVCs indicates the value of IVIVCs to the pharmaceutical industry. Because of the scientific interest and the associated utility of IVIVC as a valuable tool, the US Food and Drug Administration has published a Guidance in September 1997, entitled Extended Release Oral Dosage Forms: Development, Evaluation and Application of In Vitro/In Vivo Correlations. A predictive IVIVC enables in vitro dissolution to serve as a surrogate for in vivo bioequivalence testing. IVIVCs can be used in place of biostudies that may otherwise be required to demonstrate bioequivalence, when certain preapproval and postapproval changes are made in formulation, equipment, manufacturing process or in the manufacturing site. IVIVC development could lead to improved product quality (more meaningful dissolution specifications) and decreased regulatory burden (reduced biostudy requirements). This article will discuss in detail the FDA Guidance which deals with the development, evaluation methods and criteria, and applications of IVIVCs. From a regulatory point of view, the applications of IVIVC to grant biowaivers and to set dissolution specifications for ER oral dosage forms will be presented. Additionally, since the principles of IVIVC are considered to be similar for non oral dosage forms, the guidance for oral extended release products may be applied for non-oral products as well. While the principles are likely to be the same, it is an interesting challenge to look at appropriate methods for dissolution testing and for development of in vitro/in vivo correlations for products such as injectable depot formulations. PMID- 11389992 TI - Drug permeation enhancement via buccal route: possibilities and limitations. AB - Over the last decade, there has been a particular interest in delivering drugs, especially peptides and proteins via the buccal route. It provides direct entry into the systemic circulation thus avoiding the hepatic first-pass effect and degradation in the gastrointestinal tract, ease of administration, and the ability to terminate delivery when required. However membrane permeation can be a limiting factor for many drugs administered via the buccal route, and the epithelium that lines the oral mucosa is a very effective barrier to the absorption of drugs. In order to deliver broader classes of drugs across the buccal mucosa, reversible methods of reducing the barrier potential of this tissue must be employed. This requisite has fostered the study of penetration enhancers that will safely alter the permeability restrictions of the buccal mucosa. It has been shown that buccal penetration can be improved by using various classes of transmucosal and transdermal penetration enhancers such as bile salts, surfactants, fatty acids and derivatives, chelators, cyclodextrins and chitosan. Among these chemicals used for the drug permeation enhancement, bile salts are most common. The first part of this paper focuses on work related to the elucidation of mechanisms of action of bile salts in buccal permeation enhancement of various drugs and mucosal irritation. In the second part, results showing the enhancing effect of chitosan on buccal permeation of hydrocortisone, a commonly used topical oral anti-inflammatory agent, and transforming growth factor beta (TGF-beta), which is a bioactive peptide to which the oral mucosa is relatively impermeable is presented. PMID- 11389994 TI - Disease-induced drug targeting using novel peptide-ligand albumins. AB - Small therapeutic oligopeptides (two to 12 amino acids), designed for interaction with cytokine and growth factor receptors, unfortunately, are rapidly removed from the body. Efficient glomerular filtration and carrier-mediated membrane transport processes are involved in their clearance. By coupling of such peptides to macromolecules, elimination via these pathways is prevented and exposure to the particular receptors can be largely improved. Some of these constructs undergo receptor-mediated endocytoses and can be used as carriers to deliver associated drugs to various cell types in the body. It has been shown that, in the case of neo-glycoprotein carriers, down-regulation of the receptors aimed at can occur in the diseased state. We therefore designed a new type of polypeptide carrier, homing on receptors that are known to be highly upregulated in the pathological target tissue. For this purpose we designed ligand peptides (minimized proteins) representing the receptor-recognizing domains of PDGF and collagen type VI, aimed at receptors that are highly expressed, particularly on activated hepatic stellate cells (HSC). This myofibroblast-type of cell largely contributes to connective tissue expansion during liver fibrosis. Drug carriers for the stellate cell have not been reported before. METHODS: Cyclic octapeptide moieties (n10--12) with affinity for the two receptors were coupled to HSA (pPB HSA and pCVI-HSA, respectively). Receptor binding experiments confirmed binding of these ligand peptides to their receptors in vitro. IN VITRO STUDIES: rat HSC were isolated and purified according to standard techniques. The cells were cultured for 2 days (quiescent phenotype) or for 10 days (activated phenotype). Cell cultures were incubated with the carriers and the binding (at 4 degrees C), uptake (at 37 degrees C), and degradation were determined with radioactive and immunohistochemical methods. The results were compared with data obtained with unmodified HSA. IN VIVO STUDIES: the organ distribution of pCVI-HSA and pPB-HSA was determined 10 min after i.v. injection of tracer doses in normal and fibrotic rats, 3 weeks after bile duct ligation. Hepatocellular distribution was scored after double-immunostaining of the liver sections with an antibody against the designated hepatic cell type in combination with anti-HSA IgG. RESULTS: IN VITRO STUDIES: All three carriers preferentially bound to the activated rather than to quiescent HSC. Binding to cells was inhibitable by an excess of unlabelled pCVI HSA, endocytosis was inhibitable by 2 mM monensin suggestive of lysosomal routing of the proteins, whereas pPB-HSA, at least partly, remained at the cell surface. Degradation products of the carriers were detected extracellularly after incubation with fibrotic rat liver slices during 2-h experiments. IN VIVO STUDIES: 62+/-6% of the dose of pCVI-HSA accumulated in fibrotic livers at 10 min after injection, of which the major part was taken up in HSC. 48+/-9% of pPB-HSA accumulated in fibrotic rat livers and this carrier was also mainly taken up by HSC (5). Similar amounts of both constructs were taken up in normal rat livers, but predominantly in other cell types. The preferential homing to the stellate cells, only in the fibrotic liver is explained by the marked proliferation of this cell type as well as overexpression of the targeted receptors on these cells in the diseased state. CONCLUSIONS: The in vivo results support the in vitro studies showing accumulation of these modified albumins in HSC in fibrotic rat livers and, in particular, in the stellate cells. The results demonstrate the specificity of the stellate cell targeting and imply applicability of pCVI-HSA as carriers for drugs that act intracellularly. In addition, pPB-HSA may be used to deliver drugs that act extracellularly, such as receptor antagonists. This concept may create new opportunities for delivery of conventional drugs that are not effective enough in vivo and/or display serious extrahepatic side-effects. Minimized proteins attached to soluble or particle type of macromolecules represent a novel carrier modality of which selective body distribution is induced by the disease process to be targeted. They can be utilized as receptor antagonists and at the same time can deliver therapeutic agents to the desired site of action (dual targeting). PMID- 11389993 TI - ATTEMPTS: a heparin/protamine-based prodrug approach for delivery of thrombolytic drugs. AB - The aim of this study is to develop a heparin/protamine-based prodrug system for the controlled delivery of enzyme such as tissue-type plasminogen activator (tPA). This approach, termed antibody targeted, triggered, electrically modified prodrug-type strategy (ATTEMPTS), would permit antibody-directed administration of inactive tPA, and allow a subsequent triggered release of the active tPA at the target site. Cation-modified tPA (mtPA) was attached to a heparin--antifibrin complex via ionic interaction. The active tPA can be subsequently released by the addition of protamine, a competitive heparin inhibitor. Anti-fibrin IgG was conjugated to heparin via an end-point attachment to form the heparin--antifibrin -complex which provides the targeting efficiency of the final heparin--mtPA complex. Cation-modification was performed either by chemical conjugation by linking (Arg)(7)Cys to tPA with N-succinimidy-3-(2-pyridyldithio) propionate or by recombinant DNA method. Results show that the chemical modification process did not significantly alter specific activity of tPA with regard to plasminogen activation, fibrin-binding ability, and response toward fibrinogen. Expressed modified tPA (EmtPA) produced by recombinant DNA methods retained the same catalytic activity of the parent tPA, as well as a dynamic catalytic behavior depending upon the presence of heparin and protamine. Both types of modified tPA, especially the mtPA demonstrated a significantly higher affinity toward heparin or heparin--antifibrin complex than native tPA. In addition, the complexes of mtPA--heparin did not yield any intrinsic clot lysis activity owing to the blockage of the active site of tPA by attached heparin. On the other hand, heparin-induced inhibition of both mtPA and EmtPA activity was reversed by adding protamine, as confirmed by chromogenic and in vitro clot lysis assays. These results suggested that a heparin/protamine-based tPA delivery system may be a useful tool to improve current thrombolytic therapeutic status, by both precisely regulating the release of active tPA and aborting the associated bleeding risk. Alternatively, this ATTEMPTS approach could also be used to deliver enzyme drugs while diminishing their associated toxic effects. PMID- 11389995 TI - Tumor targeting with surface-shielded ligand--polycation DNA complexes. AB - Incorporation of the receptor binding ligands transferrin (Tf) or epidermal growth factor (EGF) into DNA/polyethylenimine (PEI) complexes was found to enhance gene transfer into tumor cell lines in a receptor-dependent manner. In systemic applications, the surface charge of DNA complexes dominated the in vivo characteristics of gene transfer. Administration of surface-shielded Tf polycation/DNA complexes into the tail vein of A/J mice resulted in preferential gene delivery into distantly growing subcutaneous Neuro2a tumors. In contrast, application of positively charged DNA/PEI complexes directed gene transfer primarily to the lung. Two alternatives of masking the surface charge of complexes were accomplished. In the first case, shielding was obtained by covalently coating of DNA/Tf-PEI complexes with polyethylene glycol (PEG). Alternatively, incorporation of sufficient Tf protein into the DNA complexes resulted in charge shielding even without PEGylation. In the latter case lower molecular weight polycations (25 kDa PEI for Tf-PEI complexes, or 32 kDa polylysine for AVET complexes) were used. PMID- 11389996 TI - Anti-JL1 antibody-conjugated poly (L-lysine) for targeted gene delivery to leukemia T cells. AB - We have designed the gene delivery carrier targeted to Molt 4 cells, human leukemia T cells, using monoclonal antibody against leukemia-specific JL1 antigen, anti-JL1 antibody, as a targeting moiety. Anti-JL1 antibody has been proven to bind to JL1 antigen and subsequently be internalized into Molt 4 cells, demonstrating that anti-JL1 antibody has the potential as a targeting ligand for leukemia-specific gene transfer. Anti-JL1 antibody was modified with the heterobifunctional crosslinker, PDPH, at carbohydrate sites and conjugated to thiolated poly-L-lysine (PLL) via disulfide bridges. The composition and antigen binding affinity of antibody-PLL conjugates were analyzed by the amino acid analysis and the flow cytometry, respectively. Antibody-PLL conjugates neutralized pSV-beta-galactosidase plasmid DNA at 5:1 weight ratio and condensed into about 200--300-nm complexes. DNA/antibody-PLL complexes were effectively internalized into Molt 4 cells after 4 h incubation at 37 degrees C and showed significantly higher in vitro transfection efficiency than DNA/PLL complexes and DNA/Lipofectin formulation due to the targeting effect of receptor-mediated endocytosis induced by anti-JL1 antibody. PMID- 11389997 TI - TerplexDNA gene carrier system targeting artery wall cells. AB - The development of non-viral gene carrier systems becomes more urgent and important due to the major biosafety considerations involved with application of viral vector systems for clinical gene therapy. We recently developed a novel non viral gene carrier system, termed TerplexDNA, which showed high gene transfer efficiency when compared to the lipofectamine gene delivery system both in HepG2 and A7R5 cell lines in vitro. In present studies, we demonstrated that the TerplexDNA gene carrier system specifically delivered the reporter genes (LacZ and Luciferase) and therapeutic gene (hrVEGF(165) cDNA) into bovine aortic artery wall cells (endothelial cells and smooth muscle cells) by receptor mediated endocytosis. We found that the transfection efficiency to these primary artery wall cells, when mediated by the TerplexDNA system, was dose-dependent, saturable and was significantly inhibited by excess free LDL. The transfection efficiency of the TerplexDNA gene carrier system was approximately 60-fold higher than that of the lipofectamine gene carrier system. The TerplexDNA gene carrier system is a useful and promising tool for artery wall gene transfer. PMID- 11389998 TI - In vivo evaluation of polymeric micellar paclitaxel formulation: toxicity and efficacy. AB - Although the current clinical formulation of paclitaxel (Taxol) has a promising clinical activity against a wide variety of tumors, it has significant toxic side effects, some of which are associated with its formulation in a 1:1 (v/v) mixture of Cremophor EL and dehydrated alcohol. One of the problems associated with the intravenous administration of paclitaxel is its low solubility in water. Our study was designed to evaluate the pharmacokinetics, tissue distribution, toxicity and efficacy of a paclitaxel (Genexol)-containing biodegradable polymeric micellar system (Genexol-PM) in comparison to Taxol. Genexol-PM was newly developed by using a low molecular weight, nontoxic and biodegradable amphiphilic diblock copolymer, monomethoxy poly(ethylene glycol)-block-poly(D,L lactide) (mPEG-PDLLA) and paclitaxel (Genexol, Samyang Genex Co., Seoul, Korea). In a human cancer cell line model, Genexol-PM and Taxol showed comparable in vitro cytotoxicity against human ovarian cancer cell line OVCAR-3 and human breast cancer cell line MCF7. The maximum tolerated dose (MTD) of Genexol-PM and Taxol in nude mice was determined to be 60 and 20 mg/kg, respectively. The median lethal dose (LD(50)) in Sprague--Dawley rats was 205.4 mg/kg (male) and 221.6 mg/kg (female) for Genexol-PM, while 8.3 mg/kg (male) and 8.8 mg/kg (female) for Taxol. After intravenous administration of Genexol-PM in murine B16 melanoma induced female SPF C57BL/6 mice at a dose of 50 mg/kg, the area under the plasma concentration-time curve (AUC) was similar to Taxol((R)) at a dose of 20 mg/kg, but biodistribution of paclitaxel after administration of Genexol-PM showed 2 to 3-fold higher levels in tissues including liver, spleen, kidneys, lungs, heart and tumor as compared to Taxol. The in vivo antitumor efficacy of Genexol-PM as measured by reduction in tumor volume of SKOV-3 human ovarian cancer implanted in nude (nu/nu) athymic mice and MX-1 human breast cancer implanted in Tac:Cr:(NCr) nu athymic mice was significantly greater than that of Taxol. The results of cytotoxicity, MTD, LD(50) and antitumor efficacy suggest that Genexol-PM may have a great advantage over present-day chemotherapy with Taxol. PMID- 11389999 TI - Biodegradable block copolymers for delivery of proteins and water-insoluble drugs. AB - Release of several drugs from new ABA-type biodegradable thermal gels, ReGel, including proteins and conventional molecules, are presented. These are biodegradable, biocompatible polymers that demonstrate reverse thermal gelation properties. Organic solvents are not used in the synthesis, purification, or formulation of these polymers. The unique characteristics of ReGel hinge on the following two key properties: (1) ReGel is a water soluble, biodegradable polymer at temperatures below the gel transition temperature; (2) ReGel forms a water insoluble gel once injected. This is consistent with a hydrophobically bonded gel state where all interactions are physical, with no covalent crosslinking. An increase in viscosity of approximately 4 orders of magnitude accompanies the sol- gel transition. The gel forms a controlled release drug depot with delivery times ranging from 1 to 6 weeks. ReGel's inherent ability to solubilize (400 to >2000 fold) and stabilize poorly soluble and sensitive drugs, including proteins is a substantial benefit. The gel provided excellent control of the release of paclitaxel for approximately 50 days. Direct intratumoral injection of ReGel/paclitaxel (OncoGel) results in a slow clearance of paclitaxel from the injection site with minimal distribution into any organ. Efficacies equivalent to maximum tolerated systemic dosing were observed at OncoGel doses that were 10 fold lower. Data on protein release (pGH, G-CSF, insulin, rHbsAg) and polymer biocompatibility are discussed. PMID- 11390000 TI - Improvements in protein PEGylation: pegylated interferons for treatment of hepatitis C. AB - Poly(ethyleneglycol) or PEG has proven to be of great value for a range of biomedical applications. A review the properties of PEG that lead to these applications is reported. Emphasis is placed on pharmaceutical uses of PEG- proteins, with specific discussion of the attributes of PEGylated alpha interferon for treatment of hepatitis C. In this latter case the choice of PEG reagent is critical to the properties of the drug, and therefore a brief presentation of PEG reagents for protein PEGylation will be given. PEGylation chemistries can be divided into first- and second-generation approaches. The first-generation chemistries are generally restricted to low-molecular-weight methoxy-PEGs because of the problem of diol contamination and resulting difunctional reagents. Problems with weak linkages and side reactions are also encountered. Second-generation PEGylation reagents avoid weak linkages and side reactions. Also they can be purified to remove diol contaminants, and as a consequence, high-molecular-weight PEGs can be used. These relatively simple chemical advances have given new vigor to PEGylation as a technology. The benefits of using high-molecular-weight, second-generation PEG reagents are demonstrated by using PEG--alpha-interferon as an example. In this case it is observed that a greatly improved drug is provided for treatment of hepatitis C. PMID- 11390003 TI - Best supportive care in non-small cell lung cancer: is there a role for radiotherapy and chemotherapy? AB - Best Supportive Care (BSC) is the treatment of choice when cure is not achievable with anticancer treatments and involves management of disease-related symptoms. In the palliative treatment of non-small cell lung cancer (NSCLC) radiation therapy has for a long time been the cornerstone of symptom management, although the best schedule is still to be defined. Chemotherapy, on the other hand, has been excluded from classical definitions of BSC and has been reserved only for selected patient populations in which a survival benefit was demonstrated using cisplatin-based regimens. We reviewed randomized trials on both palliative radiotherapy and chemotherapy in order to assess the impact of anticancer treatments on quality of life in advanced NSCLC patients. While no randomized trials compared radiation therapy with a control arm not including it, several randomized trials assessed the use of different schedules. Hypofractionated schedules seem to have comparable palliative activity when compared with the standard fractionated regimens, at least in metastatic, poor-prognosis patients. In locally advanced, inoperable NSCLC higher radiation doses administered with conventional fractionation achieve better results in terms of local control and survival. The rate of palliation of local symptoms is high, being 60-80% for chest pain and hemoptysis, while breathlessness and cough are controlled at a somewhat lower rate (50-70%). General symptoms (fatigue, anorexia, and depression) are affected in a minority of patients. Chemotherapy was compared with BSC in several randomized trials, in some of which an analysis of the quality of life was included. Results are consistent in favor of its palliative role and, when local symptom control is assessed, rates of palliation seem similar to those achieved by radiation. Benefits apply to metastatic NSCLC patients with good performance status, low body weight loss, age below 70-75. However, some studies support the use of chemotherapy also in patients with poor prognostic features. A comparison in terms of quality of life and symptom palliation between different chemotherapy regimens is the object of few trials. Both chemotherapy and radiation have an important role in the palliative treatment of advanced NSCLC patients and should be included in BSC programs. Future randomized trials should assess the best way of combining these two approaches. PMID- 11390004 TI - Potential cost-effectiveness of one-time screening for lung cancer (LC) in a high risk cohort. AB - The development of low-dose helical computed-tomography (CT) scanning to detect nodules as small as a few mm has sparked renewed interest in lung cancer (LC) screening. The objective of this study was to assess the potential health effects and cost-effectiveness of a one-time low-dose helical CT scan to screen for LC. We created a decision analysis model using baseline results from the Early Lung Cancer Action Project (ELCAP); Surveillance, Epidemiology and End Results (SEER) registry public-use database; screening program costs estimated from 1999 Medicare reimbursement rates; and annual costs of managing cancer and non-cancer patients from Riley et al. (1995) [Med Care 1995;33(8):828-841] and Taplin et al. (1995) [J Natl Cancer Inst 1995;87(6):417-26]. The main outcome measures included years of life, cost estimates of baseline diagnostic screening and follow up, and cost-effectiveness of screening. We found that in a very high-risk cohort (LC prevalence of 2.7%) of patients between 60 and 74 years of age, a one-time screen appears to be cost-effective at $5940 per life year saved. In a lower risk general population of smokers (LC prevalence of 0.7%), a one-time screen appears to be cost-effective at $23100 per life year. Even when a lead-time bias of 1 year is incorporated into the model for a low risk population, the cost effectiveness is estimated at $58183 per life year. Based on the assumptions embedded in this model, one-time screening of elderly high-risk patients for LC appears to be cost-effective. PMID- 11390005 TI - Hybrid 3D visualization of the chest and virtual endoscopy of the tracheobronchial system: possibilities and limitations of clinical application. AB - OBJECTIVE: A hybrid rendering method which combines a color-coded surface rendering method and a volume rendering method is described, which enables virtual endoscopic examinations using different representation models. MATERIALS AND METHODS: 14 patients with malignancies of the lung and mediastinum (n=11) and lung transplantation (n=3) underwent thin-section spiral computed tomography. The tracheobronchial system and anatomical and pathological features of the chest were segmented using an interactive threshold interval volume-growing segmentation algorithm and visualized with a color-coded surface rendering method. The structures of interest were then superimposed on a volume rendering of the other thoracic structures. For the virtual endoscopy of the tracheobronchial system, a shaded-surface model without color coding, a transparent color-coded shaded-surface model and a triangle-surface model were tested and compared. RESULTS: The hybrid rendering technique exploit the advantages of both rendering methods, provides an excellent overview of the tracheobronchial system and allows a clear depiction of the complex spatial relationships of anatomical and pathological features. Virtual bronchoscopy with a transparent color-coded shaded-surface model allows both a simultaneous visualization of an airway, an airway lesion and mediastinal structures and a quantitative assessment of the spatial relationship between these structures, thus improving confidence in the diagnosis of endotracheal and endobronchial diseases. CONCLUSIONS: Hybrid rendering and virtual endoscopy obviate the need for time consuming detailed analysis and presentation of axial source images. Virtual bronchoscopy with a transparent color-coded shaded-surface model offers a practical alternative to fiberoptic bronchoscopy and is particularly promising for patients in whom fiberoptic bronchoscopy is not feasible, contraindicated or refused. Furthermore, it can be used as a complementary procedure to fiberoptic bronchoscopy in evaluating airway stenosis and guiding bronchoscopic biopsy, surgical intervention and palliative therapy and is likely to be increasingly accepted as a screening method for people with suspected endobronchial malignancy and as control examination in the aftercare of patients with malignant diseases. PMID- 11390006 TI - Diagnostic results before and after introduction of autofluorescence bronchoscopy in patients suspected of having lung cancer detected by sputum cytology in lung cancer mass screening. AB - For the purpose of early detection, we have conducted population-based mass screening for lung cancer by sputum cytology since 1982. Although detection of lung cancer in its early stage is important for a good prognosis, it is often difficult to localize lesions in roentgenographically occult cancer. To clarify the role of autofluorescence bronchoscopy in localizing tumors in patients with roentgenographically occult cancer, we analyzed our diagnostic results. Fifty patients who had been detected by sputum cytology were screened by the light induced fluorescence endoscope (LIFE)-Lung System from November 1997 to April 1999. We compared the results according to the screening methods: conventional bronchoscopy alone versus LIFE with conventional white-light bronchoscopy (November 1997 to April 1999). Twenty-eight cancerous lesions and 39 borderline lesions were detected by LIFE. Of the 39 borderline lesions, nine were detected only by LIFE. Multicentric lesions including cancer or dysplasia were also detected in 21 of the 50 patients by LIFE. The sensitivity by white-light bronchoscopy alone was 85.3%, whereas that of the LIFE-Lung System with white light bronchoscopy was 94.1% (P=0.078). There were no cancerous lesions in the area observed as normal by LIFE. We also compared the diagnostic results of two localization methods: brushing of all bronchi (September 1986 to December 1990) and the LIFE-Lung System (November 1997 to April 1999). Although this was a historical comparison, the number of detected borderline lesions increased, which led to a high detection rate in patients with suspected-positive sputum (P=0.0006) by the LIFE-Lung System. In conclusion, the LIFE-Lung System is a safe and non-invasive system for detecting small intraepithelial lesions of the tracheobronchial tree. Autofluorescence bronchoscopy is more efficacious for localizing intraepithelial lesions and places fewer burdens on the patient than brushing of all bronchi. PMID- 11390007 TI - Lung cancer in patients under age 40. AB - A retrospective review of patients <40 years (n=91) seen at the Dana-Farber Cancer Institute and Brigham and Women's Hospital from January 1, 1983-January 1, 1993 was carried out. Of 91 patients, there were 43 men and 48 women with a median age of 36 years (range 28-39). Eighty percent of patients were cigarette smokers for a median of 25 pack years (range 2-68). Ninety-one percent were symptomatic at presentation. The ECOG performance status (PS) was 0 or 1 in 83%. At the time of diagnosis 15% had stage I/II, 17% stage IIIA, 22% stage IIIB and 45% stage IV disease. The most common histopathology was adenocarcinoma (46%), followed by small cell carcinoma (14%), squamous cell carcinoma (12%), large cell undifferentiated (8%) and other types (20%). The median survival for all 91 patients was 1 year with 2 and 5 year survivals of 30% and 18% respectively. Five year survival was related to stage of disease: 60% for patients with stage I, 58% for stage II, 36% for stage IIIA, 10% for stage IIIB, and 3% for stage IV disease. Factors that had no significant effect on overall survival included gender, histologic subtype, degree of differentiation, presence or absence of symptoms, and sites of metastases. Factors that adversely affected survival by univariate analysis included advanced stage of disease, poor PS, duration of symptoms for more than 3 months, and 5% or greater body weight loss. By multivariate analysis only stage (P<0.001) and weight loss (P=0.02) affected survival. This data plus results of other published studies show that young patients under age 40 with lung cancer, compared to the more common older patients, have an increased percentage of women, have a longer duration of symptoms, more often have adenocarcinoma with lower frequency of squamous cell carcinoma and sometimes small cell carcinoma, and more often present with advanced disease. Despite these differences, overall patient survival remains poor and is similar to that of older patients. PMID- 11390008 TI - Primary lung tumors in mice as an aid for understanding, preventing, and treating human adenocarcinoma of the lung. AB - Primary lung tumors in mice have morphologic, histogenic, and molecular features similar to human lung adenocarcinoma, and in particular, the bronchiolo-alveolar carcinoma subtype. Because of this, and because of the genetic homology between man and mouse and the ease of genetic manipulations in mice, this model system is receiving intense research attention. This review is intended to be informative to clinical investigators, and describes features of this model, how it is being used for translational research, and points out additional avenues of study that could have practical benefits, such as application for identifying novel therapeutic strategies. PMID- 11390009 TI - A Phase II study of docetaxel and carboplatin in the treatment of non-small cell lung cancer. AB - We investigated the efficacy of docetaxel (D) in combination with carboplatin (C) in the treatment of non-small cell lung cancer (NSCLC) patients. Since 1996, 123 with inoperable NSCLC were enrolled in the study; 120 (108 males, 12 females; mean age 58.0+/-8.3 years) were evaluated. Of those, 46 patients had squamous carcinoma, 44 adenocarcinoma, 11 large cell carcinoma and 19 undifferentiated tumours. Eligibility criteria included, documented inoperable NSCLC, WHO performance status (PS) 0-1, age up to 70 years, and normal renal and hepatic function. A total of 622 cycles of chemotherapy (CHT) (median 7 (95% CI 6.2 7.47), courses per patient) were administered. Each cycle consisted of 100 mg/m(2) of docetaxel in a 2-h infusion with C at a dose of area under the curve (AUC) of 6 on day 1. This regimen was repeated every 28 days up to eight cycles. Of the patients, five (4%) achieved complete response, 49 (40%) partial response, 47 (39%) had stable disease and 19 (15%) had progressive disease. The median survival was 12 months for all patients, 12 for the four patients with stage IIb disease, 18 for the patients with stage IIIa disease, 20 for the 29 patients with stage IIIb disease, and 11 for the 65 stage IV patients. The median time to progression was 8 months (90 patients). Toxicity was, grade 3/4 neutropenia, 18 patients (15%); grade 3/4 anaemia, 6 patients (5%); and tolerable peripheral neuropathy, 16 patients (13.3%). Responders received radiotherapy (total dose, 50 Gy in 4 weeks) between the 6th and 8th cycle. Among responders with initial stage IIIb disease, 7 (5%) underwent surgical resection. Patients with early progression of the disease received the same dose of radiotherapy between 2nd and 3rd cycle. The study is ongoing, and six patients (5%) are still alive (after 3 years). Preliminary results indicate that the D/C combination is very active in the treatment of NSCLC with tolerable toxicity. It appears that this drug combination is also good as neoadjuvant therapy in inoperable NSCLC patients. PMID- 11390010 TI - Genomic structure of the human MAD2 gene and mutation analysis in human lung and breast cancers. AB - Some of the many human cancers that exhibit chromosomal instability also carry mutations in mitotic checkpoint genes and/or reveal reduced expression of some of those genes, such as hMAD2. To facilitate investigation of alterations of hMAD2, we determined its genomic structure and intronic primers designed to amplify the entire coding region. Since general impairment of the mitotic checkpoint is frequently reported in lung cancers, and reduced expression of hMAD2 has been reported in breast cancers as well, we searched for mutations throughout the coding sequence of this gene in the genomic DNA of 30 primary lung tumors, 30 lung-cancer cell lines and 48 primary breast cancers. Our approach, which involved polymerase chain reaction-single strand conformation polymorphism (PCR SSCP) analysis and direct sequencing, revealed nucleotide variants in only two of the 108 specimens. One was a cytosine-to-adenine substitution 3 bp upstream of exon 4 that occurred in one lung cancer cell line and one primary breast tumor, a change that did not alter transcriptional sequence. The other was an adenine-to guanine substitution within exon 4, of the same lung cell line; this change already had been reported as a polymorphism. The results suggested that the hMAD2 gene is not commonly mutated in either lung nor breast cancers. Further studies should focus on other mechanisms that might account for reduced expression of the hMAD2 gene, and/or pursue analyses of other mitotic checkpoint genes for mutations in human cancer. Nevertheless, the genomic structure, the intronic primer sequences, and polymorphisms of the hMAD2 gene presented here will facilitate future studies to determine the full spectrum and frequency of the genetic events that can affect expression of the hMAD2 gene in human tumors. PMID- 11390011 TI - High syndecan-1 expression is associated with favourable outcome in squamous cell lung carcinoma treated with radical surgery. AB - Expression of syndecan-1 is down-regulated in many cellular transformation models. We studied the clinical significance of syndecan-1 expression in 116 squamous cell lung carcinomas treated with radical surgery. Paraffin-embedded tissue samples were immunostained with two antibodies against human syndecan-1 (B B4 and 104-9). Syndecan-1 expression was higher in well differentiated cancers than in moderately or poorly differentiated cancers with either antibody (P=0.001 for B-B4, and P<0.0001 for 104-9), but no significant association was found with the primary tumour size (T-stage) or the clinical stage. When the median expression (10% of cancer cells positive in B-B4 staining) was used as the cut off value, cancers with high expression were associated with more favourable survival than those with low expression (the 2-year survival rate corrected for intercurrent deaths 84% vs 61%, P=0.026). However, syndecan-1 expression was not an independent prognostic factor in a multivariate survival analysis. We conclude that syndecan-1 expression decreases in parallel with histological dedifferentiation in squamous cell carcinoma of the lung, and that low syndecan-1 expression is associated with unfavourable outcome. PMID- 11390012 TI - Longitudinal study of resting energy expenditure, body cell mass and the inflammatory response in male patients with non-small cell lung cancer. AB - The aim of this study was to examine the inter-relationship between the inflammatory response and resting energy expenditure in patients with non-small cell lung cancer (NSCLC) before and after the onset of weight loss. Healthy subjects (n=7) and patients with NSCLC without weight loss (n=12) were studied. Resting energy expenditure adjusted for metabolically active tissue, as measured by total body potassium, was approximately 15% higher in the NSCLC group (P<0.01). Moreover, the resting energy expenditure, correlated with the magnitude of the inflammatory response (r=0.753, P<0.01). Six cancer patients subsequently lost weight and the relationship between resting energy expenditure and the inflammatory response was maintained. These results highlight the impact of the inflammatory response on the increase in the resting energy expenditure which precedes the onset of weight loss in patients with NSCLC. PMID- 11390013 TI - Retro-inverso peptide analogues of Trypanosoma cruzi B13 protein epitopes fail to be recognized by human sera and peripheral blood mononuclear cells. AB - Retro inverso (RI) analogues of antigenic synthetic peptides, which are made of D amino acids with a reversed sequence, may mimic the side chain conformation of natural all-L peptides. RI analogues were cross-reactively recognized by antibodies and CD4+ T cells reactive against natural all-L synthetic peptides or native proteins in animal models. Since peptides containing D-amino acids are highly resistant to proteolytic digestion, cross-reactive RI analogues may be ideal for in vivo administration to humans as synthetic peptide vaccines or immunomodulators. B13 is an immunodominant tandemly repetitive protein from Trypanosoma cruzi, a protozoan parasite that is the causative antigen of Chagas' disease. In order to test whether RI peptides can be recognized by human antibody and T cells, we synthesized two all-L peptides containing the immunodominant B (S12) and T (S15.7) cell epitopes of B13 protein from T. cruzi and their retro (R, made of all-L amino acids with reversed sequence), inverso (I, made of all-D amino acids) and RI analogues. Recognition of peptides S12, S12-R, S12-I and S12 RI by anti-B13 antibodies in sera from T. cruzi-infected patients was tested in competitive ELISA assay with recombinant B13 protein as the solid phase antigen. Peptides S15.7 and its topological analogues were tested at the 10-50 microM range in proliferation assays on peripheral blood mononuclear cells (PBMC) from S15.7-responder individuals. The median percentage inhibition of B13 ELISA for peptide S12 was 94%, while those of the RI analogue or the other topological analogues were below 12%. While peptide S15.7 was recognized by PBMC from all subjects tested, none recognized the RI analogue of the S15.7 T cell epitope. Our results indicate that cross-reactivity with natural epitopes is not an universal property of RI analogues. This may limit the general applicability of the use of cross-reactive RI analogues as human vaccines and immunotherapeutic agents. PMID- 11390014 TI - Characterization of the pharmacology, signal transduction and internalization of the fluorescent PACAP ligand, fluor-PACAP, on NIH/3T3 cells expressing PAC1. AB - Fluor-PACAP, a fluorescent derivative of PACAP-27, has been confirmed to share a high affinity for PAC1 receptors transfected into NIH/3T3 cells and to have comparable pharmacological characteristics to the unconjugated, native form. Through competitive binding with 125I-PACAP-27, the two ligands exhibited similar dose- dependent inhibition. Additional examination of the efficacy of activating adenylyl cyclase revealed that both ligands analogously stimulated the production of cyclic AMP. Furthermore, PAC1 internalization visualized by our Fluor-PACAP, is compareable to that performed with the radioligand, 125I-PACAP-27, with maximal internalization achieved within thirty minutes. Thus, Fluor-PACAP exhibits intracellular signaling abilities homologous to the native ligand. PMID- 11390015 TI - The neuromodulatory effects of VIP/PACAP on PC-12 cells are associated with their N-terminal structures. AB - ONOUE, S., WAKI, Y., NAGANO, Y., SATOH, S., KASHIMOTO, K. Neuromodulatory Effects of VIP/PACAP on PC-12 Cells Are Associated with Their N-terminal Structures. PEPTIDES xx(xx) 000-000, 200x.- The current study explored whether the differences in biological activities in PC-12 cells between vasoactive intestinal polypeptide (VIP) and pituitary adenylate cyclase-activating polypeptide (PACAP) are attributable to the sequence difference in their N-terminal portions and are correlated with the solution structures of the peptides. In the neurite outgrowth assay, N-terminal modification of VIP to PACAP-like sequences altered its effect, the activity was confirmed even at a low concentration (10(-10) M). On the contrary, N-terminal modification of PACAP 27 to VIP-like sequences reduced its activity. These relationships were also confirmed for the inhibitory effects of the peptide analogues on PC-12 cells growth at 10(-7) M. The present results combined with our previously reported data, including binding assay, support that the N-termini of VIP/PACAP plays an important role in their activities. PMID- 11390016 TI - Pituitary adenylate cyclase activating polypeptide is highly abundant in the nervous system of anoxia-tolerant turtle, Pseudemys scripta elegans. AB - The levels of the pituitary adenylate cyclase activating polypeptide (PACAP) were measured in the central nervous system and in peripheral organs of the anoxia tolerant freshwater turtle, Pseudemys scripta elegans by radioimmunoassay. The concentration of PACAP38 was strikingly high in the central nervous system and lower but considerable immunoreactivity was detected in the peripheral organs. Levels of PACAP38 in the turtle brain exceed those measured in rat and human brain areas by 10-100-fold. Based on these exceptionally high levels of PACAP and the known neuroprotective role of the peptide, it can be suggested that PACAP38 plays a role in the extraordinary resistance of the turtle brain from anoxia induced neuronal damage. PMID- 11390017 TI - Antiproliferative actions of growth hormone-releasing hormone antagonists on MiaPaCa-2 human pancreatic cancer cells involve cAMP independent pathways. AB - We evaluated the effects of GHRH antagonists on the proliferation of MiaPaCa-2 human pancreatic cancer cells and cAMP signaling in vitro. GHRH antagonists inhibited the proliferation of MiaPaCa-2 cells in vitro in a dose-dependent way and caused a significant elevation in cAMP production. In a superfusion system, short-term exposure of the cells to GHRH antagonists evoked an acute, dose dependent release of cAMP into the medium. Native GHRH, which stimulates cAMP efflux from pituitary at nanomolar doses, did not influence cAMP release from cultured or superfused MiaPaCa-2 cells even at 10-30 microM. VIP, PACAP, secretin and glucagon also did not influence cell proliferation or cAMP production. Adenylate cyclase activator forskolin (FSK) caused a greater cAMP response, but a smaller antiproliferative effect than GHRH antagonists. Combined treatment with FSK and GHRH antagonist JV-1-38 potentiated the cAMP-inducing effect of FSK, but did not produce a greater inhibition of cell proliferation than JV-1-38 alone. A selective accumulation of radiolabeled GHRH antagonist [(125)I]JV-1-42 in vivo in MiaPaCa-2 carcinoma xenografted into nude mice was also observed. In conclusion, second messengers other than cAMP participate in the signal transduction pathways of GHRH analogs mediated by tumoral GHRH receptors. PMID- 11390018 TI - Blockade of pancreatic polypeptide-sensitive neuropeptide Y (NPY) receptors by agonist peptides is prevented by modulators of sodium transport. Implications for receptor signaling and regulation. AB - Ligand binding to rodent pancreatic polypeptide-responding neuropeptide Y (NPY) receptors (here termed PP/NPY receptors), or to cloned Y4 or Y5 receptors, is selectively inhibited by amiloride, peptide or alkylating modulators of sodium transport. The PP/NPY and Y4 receptors are also selectively blocked by human or rat pancreatic polypeptide (PP) and the blocking peptides are not dissociated by high concentrations of alkali chlorides (which restore most of the binding of subtype-selective agonists to Y1 and Y2 sites). The PP/NPY receptors could also be blocked by NPY and related full-length peptides, including Y1-selective agonists (IC50 300-400 pM). The cloned Y(4) receptors from three species are much less sensitive to NPY or PYY. The sensitivity of both the PP/NPY sites and the Y(4) sites to Y2-selective peptides is quite low. The ligand attachment to PP/NPY sites is also very sensitive to peptidic Y1 antagonist ((Cys31,NVal34NPY27-36))2, which however blocks these sites at much higher molarities. Blockade of PP/NPY and Y4 sites by agonist peptides can be largely prevented by N5-substituted amiloride modulators of Na+ transport, and by RFamide NRNFLRF.NH2, but not by Ca2+ channel blockers, or by inhibitors of K+ transport. Protection of both PP/NPY and Y4 sites against blockade by human or rat pancreatic polypeptide is also afforded by short N-terminally truncated NPY-related peptides. The above results are consistent with a stringent and selective activity regulation for rabbit PP/NPY receptor(s) that may serve to differentiate agonists and constrain signaling, and could involve transporter-like interactants. PMID- 11390019 TI - Diazoxide effects on hypothalamic and extra-hypothalamic NPY content in Zucker rats. AB - To determine if the anorectic effects of the insulin antagonist diazoxide (DZ) are mediated by reduced central neuropeptide Y (NPY), female Zucker rats, given DZ (150 mg/kg/day) or placebo for about four weeks, were sacrificed following overnight fasting or free feeding. Several hypothalamic and extra-hypothalamic nuclei were extracted for NPY content. DZ reduced weight gain in obese rats and lowered glucose of lean and obese rats without affecting insulin. Contrary to the hypothesis, DZ increased NPY in hypothalamic nuclei of free fed lean and obese rats. DZ elevated hypothalamic NPY levels in fasted obese rats and had more diverse effects in extra-hypothalamic nuclei of lean rats. PMID- 11390020 TI - Direct inhibitory effect of adrenomedullin, calcitonin gene-related peptide, calcitonin, and amylin on cholecystokinin-induced contraction of guinea-pig isolated caecal circular smooth muscle cells. AB - We recently reported the direct inhibitory effect of adrenomedullin on caecal circular smooth muscle cells via cAMP system. This study was designed to determine whether the structurally related peptides to adrenomedullin (i.e.; calcitonin gene-related peptide (CGRP), calcitonin, and amylin) can inhibit the cholecystokinin octapeptide (CCK-8)-induced contractile response by exerting a direct action on guinea-pig caecal circular smooth muscle cells, and to compare the inhibitory potency of these peptides. In addition, to elucidate each intracellular mechanisms, the effects of an inhibitor of cAMP-dependent protein kinase, inhibitors of particulate or soluble guanylate cyclase on the each peptide-induced relaxation were investigated. Adrenomedullin, CGRP, calcitonin, and amylin inhibited the contractile response produced by CCK-8 in a dose dependent manner, with IC50 values of 0.14 nM, 0.37 nM, 5.4 nM, and 160 nM, respectively. An inhibitor of cAMP-dependent protein kinase significantly inhibited the relaxation produced by all of these peptides. On the contrary, inhibitors of particulate or soluble guanylate cyclase did not have any significant effect on the relaxation produced by these peptides. In this study, we demonstrated the direct inhibitory effects of the structurally related peptides to adrenomedullin (i.e.; CGRP, calcitonin, and amylin) on the isolated caecal circular smooth muscle cells via cAMP system. The order of potency was as follows; adrenomedullin falling dots CGRP > calcitonin > amylin. PMID- 11390021 TI - Role of calcitonin gene-related peptide and kinins in post-ischemic intestinal reperfusion. AB - The involvement of kinins, calcitonin gene-related peptide (CGRP), and tachykinins during mesenteric post-ischemic reperfusion was studied in anesthetized rats by using antagonists for bradykinin (BK) B1, BK B2, CGRP1, or tachykinin NK1 receptor, or by capsaicin-induced desensitization. B1, B2, or CGRP1 receptor antagonists or desensitization attenuated the transient hypotension and plasma protein and leukocyte infiltration of intestinal wall observed during post-ischemic reperfusion. These effects were abolished by the combination of B2 and CGRP1 blockade as well as by B2 antagonism in capsaicinized rats, while NK1 blockade was ineffective. Our results suggest that kinins and CGRP contribute to systemic vasodilatation and microvascular leakage during mesenteric reperfusion. Pharmacological blockade of these systems could help preventing hypotension and intestinal injury consequent to reperfusion. PMID- 11390022 TI - Stimulation of endogenous nitric oxide production is involved in the inhibitory effect of adrenomedullin on aldosterone secretion in the rat. AB - Adrenomedullin (AM) (10(-8) M) partially suppressed aldosterone response of dispersed rat zona glomerulosa (ZG) cells to 10 mM K+, and the nitric oxide (NO) synthase inhibitors L-NAME (10(-3) M) and 1400W (10(-4) M) effectively counteracted this effect of AM. The NO donor L-Arginine (L-Arg) (10(-5) M) decreased both basal and K+ -stimulated aldosterone secretion. The guanylate cyclase inhibitor Ly-83583, at a concentration (10(-4) M) abolishing either the guanylate-cyclase activator guanylin- or L-Arg-induced cGMP release from dispersed ZG cells, did not affect the aldosterone antisecretagogue action of AM and L-Arg. AM (10(-8) M) evoked a moderate increase in cGMP release by dispersed ZG cells, and the effect was blocked by both 10(-4) M Ly-83583 and 10(-3) M L NAME. Collectively, these findings allow us (1) to confirm that NO inhibits aldosterone secretion through a cGMP-independent mechanism; and (2) to suggest that stimulation of endogenous NO synthesis plays a role in the mechanisms underlying the inhibitory effect of AM on K+ -stimulated aldosterone secretion from rat ZG cells. PMID- 11390023 TI - Gender differences in the attenuation of salt-induced hypertension by angiotensin (1-7). AB - Chronic infusion of angiotensin (1-7) [Ang-(1-7)] lowers blood pressure in spontaneously hypertensive rats (SHR). To assess the role of Ang-(1-7) in salt induced hypertension, Ang-(1-7) (24 microg/kg/hr) or saline was administered chronically via osmotic minipump into the jugular vein of 5-6 wk-old male (M) and female (F) Dahl salt-sensitive rats placed on a high-salt (8% NaCl) diet for 2 weeks. Blood pressure (BP) and heart rate were measured prior to the start of the diet and weekly thereafter. Ang-(1-7) significantly attenuated the BP increase after 1 wk on the diet in both M and F rats, but after 2 weeks only in F rats. Enhanced release of prostacyclin, (6-keto PGF1 alpha), following Ang-(1-7) treatment was observed in both M and F rats. In addition, significant increases in aortic blood flow and plasma levels of nitric oxide were observed in the F rats following Ang-(1-7) treatment. These findings demonstrate that the reduction in BP is due to both prostacyclin and NO and that there is a gender difference in the attenuation of salt-induced hypertension by Ang-(1-7). PMID- 11390024 TI - A role for the angiotensin IV/AT4 system in mediating natriuresis in the rat. AB - Angiotensin II (AngII) or Angiotensin IV (AngIV) was infused into the renal artery of anesthetized rats while renal cortical blood flow was measured via laser Doppler flowmetry. The infusion of AngII produced a significant elevation in mean arterial pressure (MAP) with an accompanying decrease in cortical blood flow, glomerular filtration rate (GFR), urine volume, and urine sodium excretion. The infusion of AngIV induced significant increases in renal cortical blood flow and urine sodium excretion, without altering MAP, GFR, and urine volume. Pretreatment infusion with a specific AT1 receptor antagonist, DuP 753, blocked or attenuated the subsequent AngII effects, while pretreatment infusion with the specific AT4 receptor antagonist, Divalinal-AngIV, blocked the AngIV effects. These results support distinct and opposite roles for AngII and AngIV, i.e. AngII acts as an anti-natriuretic agent, while AngIV acts as a natriuretic agent. PMID- 11390025 TI - Angiotensinase activity in mice fed an olive oil-supplemented diet. AB - We evaluated the influence of a diet supplemented with olive oil (20% by weight) (OO) on the activity of glutamyl aminopeptidase (GluAP) and aspartyl aminopeptidase (AspAP), which are involved in angiotensin metabolism. Serum concentrations of total cholesterol and fatty acids were also measured. Animals fed on the OO diet gained significantly more weight than did controls from the second week until the end of the feeding period. Serum total cholesterol concentration was significantly higher in the OO group than in control mice. Total monounsaturated fatty acids increased in OO-fed animals, but total saturated fatty acids decreased. No differences between the two groups were observed for total polyunsaturated fatty acids. Serum from animals fed on the OO diet contained significantly lower proportions of myristic, pentadecanoic, palmitic, palmitoleic, vaccenic, alpha-linolenic, gamma-linolenic, and 11,14 eicosadienoic acids than did serum from control animals. In contrast, the OO group had higher levels of oleic, stearic, and gondoic acids. GluAP activity decreased significantly in the serum of OO-fed animals. In these animals soluble AspAP activity was significantly higher in the testis, and significantly lower in the lung and adrenal, in comparison to controls. Membrane-bound AspAP activity was higher in the testis and atrium, and lower in lung, in the OO group. Soluble GluAP activity was significantly lower in the testis of OO-fed animals. Membrane bound GluAP activity did not differ between the two groups in any of the tissues analyzed. Serum AspAP and GluAP activities correlated negatively with palmitoleic and vaccenic acid respectively in the OO group. However, no significant correlations were found in the control group. These results may reflect functional changes in the renin-angiotensin system in the serum, adrenal, testis, lung and atrium after feeding with a diet enriched in olive oil. PMID- 11390026 TI - Effects of the vasopeptidase inhibitor omapatrilat on cardiac endogenous kinins in rats with acute myocardial infarction. AB - The purposes of this study were to evaluate and to compare the effects of simultaneous angiotensin-converting enzyme (ACE) and neutral endopeptidase 24.11 (NEP) inhibition by the vasopeptidase inhibitor omapatrilat (1 mg. kg(-1). day( 1)) with those of the selective ACE inhibitor enalapril (1 mg. kg(-1). day(-1)) on survival, cardiac hemodynamics, and bradykinin (BK) and des-Arg(9)-BK levels in cardiac tissues 24 h after myocardial infarction (MI) in rats. The effect of the co-administration of both B(1) and B(2) kinin receptor antagonists (2.5 mg. kg(-1). day(-1) each) with metallopeptidase inhibitors was also evaluated. The pharmacological treatments were infused subcutaneously using micro-osmotic pumps for 5 days starting 4 days before the ligation of the left coronary artery. Immunoreactive kinins were quantified by highly sensitive and specific competitive enzyme immunoassays. The post-MI mortality of untreated rats with a large MI was high; 74% of rats dying prior to the hemodynamic study. Mortality in the other MI groups was not significantly different from that of the untreated MI rats. Cardiac BK levels were not significantly different in the MI vehicle treated group compared with the sham-operated rats. Both omapatrilat and enalapril treatments of MI rats significantly increased cardiac BK concentrations compared with the sham-operated group (P < 0.05). However, cardiac BK levels were significantly increased only in the MI omapatrilat-treated rats compared with the MI vehicle-treated group (P < 0.01). Cardiac des-Arg(9)-BK concentrations were not significantly modified by MI, and MI with omapatrilat or enalapril treatment compared with the sham-operated group. The co-administration of both kinin receptor antagonists with MI omapatrilat- and enalapril-treated rats had no significant effect on cardiac BK and des-Arg(9)-BK levels. Thus, the significant increase of cardiac BK concentrations by omapatrilat could be related to a biochemical or a cardiac hemodynamic parameter on early (24 h) post-MI state. PMID- 11390027 TI - Muscle layer- and region-dependent distributions of oxytocin receptors in the porcine myometrium. AB - The aim of the present study was to clarify smooth muscle- and region-dependent distributions of the oxytocin receptor that mediates oxytocin-induced contraction in the nonpregnant porcine myometrium by means of mechanical and radioligand ([3H]-oxytocin) binding studies. In Krebs solution, oxytocin (0.1-300 nM) caused concentration-dependent contractions of the cornual myometrium, and the longitudinal muscle was more sensitive than the circular muscle. [Arg8] vasopressin and [deamino-Cys1, D-Arg8]-vasopressin also contracted the myometrium, and the order of the potency was oxytocin > [Arg8]-vasopressin > [deamino-Cys(1), D-Arg(8)]-vasopressin. Treatment with a high concentration of oxytocin selectively inhibited the contraction of oxytocin and [Arg8]-vasopressin without affecting the responses of acetylcholine and high-K+. Selective cross inhibition was also observed in the presence of a high concentration of [Arg(8)] vasopressin. The oxytocin-induced contraction was resistant to tetrodotoxin and atropine, but was reduced by verapamil or by the removal of external Ca2+, indicating that oxytocin has a direct action on smooth muscle cells and that extracellular Ca2+ plays an important role for the contraction. In Kumagai solution, oxytocin caused contraction of the cornual longitudinal muscle ( logEC50 = 8.5) but not the circular muscle. Longitudinal muscles of other regions (corpus and cervix) were also responsive to oxytocin, but the -logEC50 value differed from region to region (cornua > corpus = cervix). On the other hand, oxytocin failed to cause contraction of the corpus and cervical circular muscles. 3H-Oxytocin bound to crude membrane preparations of the myometrium in a concentration-dependent (0.084-2.7 nM) saturable manner. Scatchard analysis of equilibrium binding data revealed the presence of a single class of binding site with an apparent dissociation constant (Kd, 1.1-1.5 nM), but receptor density (Bmax) differed in the two muscle layer types (longitudinal muscle: circular muscle = 5:1) and tended to decrease from the cornua to the cervix. In conclusion, the receptor specific for oxytocin is present in the porcine myometrium and mediates the contractile responses of both oxytocin and [Arg8] vasopressin. The distribution of the oxytocin receptors differs according to the type of muscle layer (longitudinal muscle > circular muscle) and the region of the uterus. PMID- 11390028 TI - Chromatin-derived acidic peptides modulate catecholamine release in the hypothalamus. AB - We have studied the neuromodulatory effects of three synthetic peptides, structurally related to chromatin-derived acidic peptides (ACPs): ACP-1 (Asp-Asp Ser-Asp-Glu-Glu-Asn), corresponding to the C-terminal fragment of the largest subunit of eukaryotic RNA polymerase II; a more lipophilic derivative, ACP-2 (Ala Ile-Ser-Pro-Asp-Asp-Ser-Asp-Glu-Glu-Asn); and its phosphorylated form ACP-3 (Ala Ile-Ser-Pro-Asp-Asp-Ser(P)-Asp-Glu-Glu-Asn). Rat hypothalamic synaptosomes, loaded with [(3)H]norepinephrine or [(3)H]dopamine, were perfused with the above peptides, both basally and during a depolarizing stimulus. We have found: ACP-1 inhibited both dopamine and norepinephrine release; ACP-2 inhibited dopamine release, without affecting norepinephrine release; ACP-3 was almost ineffective, except for a weak dopamine inhibiting effect only at a higher concentration. PMID- 11390029 TI - N-Acetylcarnosine, a natural histidine-containing dipeptide, as a potent ophthalmic drug in treatment of human cataracts. AB - A study was designed to document and quantify the changes in lens clarity over 6 and 24 months in 2 groups of 49 volunteers (76 eyes) with an average age of 65.3 +/- 7.0 enrolled at the time of diagnosis of senile cataracts of minimal to advanced opacification. The patients received N-acetylcarnosine, 1% sol (NAC) (26 patients, 41 eyes = Group II), placebo composition (13 patients, 21 eyes) topically (two drops, twice daily) to the conjunctival sac, or were untreated (10 patients, 14 eyes); the placebo and untreated groups were combined into the control (reference) Group I. Patients were evaluated upon entry, at 2-month (Trial 1) and 6-month (Trial 2)-intervals for best corrected visual acuity (b/c VA), by ophthalmoscopy and the original techniques of glare test (for Trial 1), stereocinematographic slit-image and retro-illumination photography with subsequent scanning of the lens. The computerized interactive digital analysis of obtained images displayed the light scattering/absorbing centers of the lens into 2-D and 3-D scales. The intra-reader reproducibility of measuring techniques for cataractous changes was good, with the overall average of correlation coefficients for the image analytical data 0.830 and the glare test readings 0.998. Compared with the baseline examination, over 6 months 41.5% of the eyes treated with NAC presented a significant improvement of the gross transmissivity degree of lenses computed from the images, 90.0% of the eyes showed a gradual improvement in b/c VA to 7-100% and 88.9% of the eyes ranged a 27-100% improvement in glare sensitivity. Topographic studies demonstrated less density and corresponding areas of opacification in posterior subcapsular and cortical morphological regions of the lens consistent with VA up to 0.3. The total study period over 24 months revealed that the beneficial effect of NAC is sustainable. No cases resulted in a worsening of VA and image analytical readings of lenses in the NAC-treated group of patients. In most of the patients drug tolerance was good. Group I of patients demonstrated the variability in the densitometric readings of the lens cloudings, negative advance in glare sensitivity over 6 months and gradual deterioration of VA and gross transmissivity of lenses over 24 months compared with the baseline and 6-month follow-up examinations. Statistical analysis revealed the significant differences over 6 and 24 months in cumulative positive changes of overall characteristics of cataracts in the NAC-treated Group II from the control Group I. The N-acetylated form of natural dipeptide L carnosine appears to be suitable and physiologically acceptable for nonsurgical treatment for senile cataracts. PMID- 11390030 TI - Neuronal messengers in the human cerebral circulation. AB - In recent years our knowledge of the nervous control of the cerebral circulation has increased. The use of denervations and retrograde tracing in combination with immunohistochemical techniques has demonstrated that cerebral vessels are supplied with sympathetic, parasympathetic, and sensory nerve fibers and possibly central pathways containing a multiplicity of new transmitter substances in addition to the classical transmitters. The majority of these transmitters are neuropeptides. More recently it has been suggested that a gaseous transmitter, nitric oxide (NO) also could participate in the neuronal regulation of cerebral blood flow. Although little is known about the physiological actions and inter relationships among all these putative neurotransmitters, their presence within cerebrovascular nerve fibers will make it necessary to revise our view on the mechanisms of cerebrovascular neurotransmission. PMID- 11390031 TI - Spreading of HIV-specific CD8+ T-cell repertoire in long-term nonprogressors and its role in the control of viral load and disease activity. AB - Long-term non-progressors (LTNP) represent a minority of human immunodeficiency virus (HIV) infected individuals characterized by stable or even increasing CD4+ T-cell count and by stronger immune responses against HIV than progressors. In this study, HIV-specific effector CD8+ T cells, as detected by both a sensitive ex vivo enzyme-linked immunospot (ELISPOT) assay and specific major histocompatibility complex (MHC) peptide tetramers, were at a low frequency in the peripheral blood of LTNP, and recognized a lower number of HIV peptides than their memory resting cell counterparts. Both factors may account for the lack of complete HIV clearance by LTNP, who could control the viral spread, and displayed a higher magnitude of cytotoxic T lymphocyte (CTL) responses than progressors. By combining cell purification and ELISPOT assays this study demonstrates that both effector and memory resting cells were confined to a CD8+ population with memory CD45RO+ phenotype, with the former being CD28- and the latter CD28+. Longitudinal studies highlighted a relatively stable HIV-specific effector repertoire, viremia, and CD4+ T-cell counts, which were all correlated with maintenance of nonprogressor status. In conclusion, the analysis of HIV-specific cellular responses in these individuals may help define clear correlates of protective immunity in HIV infection. PMID- 11390032 TI - Regeneration and tolerance factor is expressed during T-lymphocyte activation and plays a role in apoptosis. AB - Regeneration and tolerance factor (RTF) is a protein cloned from the thymus and expressed on B lymphocytes in normal pregnancy, B lymphocytic leukemia lines, and T and B lymphocytes in individuals with HIV infection. Findings, using the Jurkat T-cell model, revealed that RTF is upregulated after activation and anti-RTF antibody-induced apoptosis. In this article anti-RTF antibody-induced apoptosis of both unstimulated and activated T lymphocytes. RTF expression was examined in human PBMC or purified T lymphocytes after their in vitro activation. Kinetic studies indicated maximal RTF cell surface expression on activated T lymphocytes occurred between expression of the early activation antigen CD69 and the IL 2alpha receptor (CD25) by multiparameter flow cytometry. RTF receptor expression correlated with Fas (CD95) and CD25 receptor expression (r2 = 0.6 and 0.5, respectively). RTF surface expression was dependent on the stimuli used to activate T lymphocytes. T lymphocytes obtained maximal RTF expression when activated through the TCR signal complex using anti-CD3epsilon antibody alone when compared with T lymphocytes activated with costimulation provided by anti CD28 antibody alone or with anti-CD28 and anti-CD3epsilon antibody. RTF is expressed under conditions of both activation and anergy. The RTFs increased concentration on the surface of anergic T cells may protect these cells from apoptosis because increased RTF concentrations inhibited anti-RTF induced apoptosis. These data further characterize the expression of RTF on activated T lymphocytes and the role of anti-RTF antibody in T-lymphocyte apoptosis. PMID- 11390033 TI - Ultraviolet-B radiation induces modulation of antigen presentation of herpes simplex virus by human epidermal cells. AB - Although ultraviolet (UV) B radiation is known to be immunosuppressive, there is little information regarding a relevant immunological endpoint to assess human subjects in vivo. Therefore, we have examined the effect of in vivo UV radiation on the ability of human epidermal cells (EC) to present herpes simplex virus (HSV) antigens to memory T cells. Human volunteers, who were seropositive for HSV, were exposed to one minimal erythemal dose (MED) for four consecutive days. EC, prepared from suction blister roofs, were co-cultured with autologous T cells in the presence of HSV. HSV antigen presentation by UV-exposed EC was increased compared with control, nonexposed EC. This up-regulation correlated with an influx of macrophages into the epidermis, which are considered to be associated with UV-induced tolerance. Altering the UV protocol to a sub-erythemal UV dose for four consecutive days or to a single high dose of 2 MED, resulted in suppressed HSV antigen presentation, without the influx of the UV-macrophages. One of the goals of the present study was to eventually use this HSV system to investigate sunscreen immunoprotection. A pilot study with a TiO2-containing sunscreen suggested that the endpoint for UV-induced immunosuppression presented here is promising to be used for human in vivo sunscreen immunoprotection studies. PMID- 11390034 TI - Noncytolytic human lymphocytes injure dermal microvessels in the huPBL-SCID skin graft model. AB - Recent transplantation experiments using perforin-deficient mice as allograft recipients have challenged the concept that allograft rejection is mediated exclusively by CTL. We sought to determine if human noncytolytic lymphocytes could mediate rejection of allogeneic human skin grafts in the huPBL-SCID mouse model of rejection. We generated short term lines of human lymphocytes from peripheral blood mononuclear cells using PHA as a mitogen. The first group was stimulated with PHA alone, the second with PHA plus IL-4 and neutralizing antibody to IL-12, and in the third group PBL were depleted of B cells and monocytes before stimulation as in group 2. After two passages, lines were tested for cytolytic ability and IFN-gamma production. Each line was injected i.p. to mice bearing allogeneic skin grafts. The grafts were harvested between day 16 and 21 after PBL injection, then the histology was scored by a blinded observer for degree of infiltration, microvessel injury, induction of epidermal MHC class II, and perforin expression. In vitro we found that PBL in groups 2 and 3 were unable to lyse cultured endothelial cells in a lectin-directed 111In release assay. In vivo 80% of the IL-4/anti-IL-12 groups maintained the IFN-gamma-low phenotype, and no perforin was detected in these grafts. Nevertheless, human microvessel injury was similar between the two groups. This was not antibody-dependent since the B-cell-depleted group showed similar injury. Moreover adjacent murine vessels were intact. We interpret these observations to show (1) these human PBL lines maintained their phenotype following in vivo restimulation, and (2) noncytolytic graft-infiltrating lymphocytes specifically promote injury of allogeneic human microvessels. PMID- 11390035 TI - In vitro co-incubation of pig islet cells with xenogeneic human blood mononuclear cells causes loss of insulin release during perifusion: involvement of non-T-cell and T-cell-mediated mechanisms. AB - Because the different steps of the human cellular immune rejection of pig islets are still poorly understood, our previous work concerned the intensity and mechanisms of the proliferation of human peripheral blood mononuclear cells (PBMC) to adult pig islet cells (PIC). As lymphocyte proliferation is not indicative of alteration of PIC, the present in vitro study evaluated cell mediated immune effectors possibly involved in impairment of adult PIC. A test was thus developed, based on perifusion analysis of the alteration of insulin release from PIC incubated with different human cells. Compared to PIC incubation alone or with autologous pig splenocytes, seven-day co-incubation with whole human peripheral blood mononuclear cells (PBMC) (n = 18) led to almost complete abolition of basal and stimulated insulin releases (p < 0.0001). This effect could not be reversed by extensive sequential washes before perifusion of PIC, and the number of PIC was decreased by 78% after seven-day co-incubation with PBMC. PBMC are a complex mixture of cells involved in different xenogeneic mechanisms, and two components of this PIC impairment were then detected separately. First, the effect of PBMC against PIC was decreased (p < 0.0001) after removal of either MHC class II+ or CD14+ cells from PBMC. On the contrary, decreasing effect (p < 0.001) on insulin secretion was observed when only plastic adherent or CD14+ cells were co-incubated with PIC. Additionally, alteration of insulin release from PIC cultured with PBMC or plastic-adherent cells was abolished dose-dependently (p < 0.0001 and p < 0.04, respectively) by gadolinium chloride (which inhibits macrophages), but not modified by cyclosporin A or mycophenolate mofetil which did not alter insulin release from PIC but blocked the proliferation of PBMC against PIC. A second mechanism was also detected, since co-incubation of PIC with purified human T cells remixed with antigen presenting cells led to a decrease (p < 0.0001) of insulin release. This model based on the alteration of dynamic basal and stimulated insulin secretion provides detailed account of in vitro human cell-mediated impairment of PIC. It shows that the xenogeneic effect of whole mononuclear cells was strong and rapid. A crucial role was played by MHC class II+, CD14+, and plastic-adherent cells. Two mechanisms appear to be responsible for the role of these cells: 1) early direct effect, potentially involved in vivo in primary nonfunction of islets aggressed by monocytes/macrophages; and 2) the presentation of PIC xenoantigens leading to impairment by T lymphocytes, which may be involved in in vivo specific cellular rejection. PMID- 11390036 TI - Clinical significance of soluble form of HLA class I molecule in Japanese patients with pancreatic cancer. AB - In recent studies a soluble form of human leukocyte antigen class I (sHLA-I) has been found in blood, urine, ascitic fluid, and various other tissues. Research has been focused on the role of sHLA-I in the induction of immunotolerance in organ transplantation. To examine the role of sHLA-I in the immune system of patients with malignancy, we examined serum sHLA-I levels in patients with pancreatic, biliary, hepatic malignancy, and other diseases. We examined sHLA-I levels in the sera of patients with pancreatic cancer (n = 19), benign biliary disease and chronic pancreatitis (n = 20), hepatocellular carcinoma (n = 51), gallbladder cancer (n = 6), cholangiocellular carcinoma (n = 6), and in normal controls (n = 22), using enzyme-linked immunosorbent assay (ELISA). In patients with pancreatic cancer we also analyzed the relationship between sHLA-I and CA19 9, and the specificity and sensitivity of sHLA-I. When patients with acute or chronic hepatitis were excluded from analysis, the mean sHLA-I level in patients with pancreatic cancer was significantly higher than that of normal controls (p < 0.01) and patients with benign disease (p < 0.01), hepatocellular carcinoma (p < 0.01), gallbladder cancer (p < 0.05), and cholangiocarcinoma (p < 0.05). We determined a serum sHLA-I cutoff level for normal controls of 2000 ng/ml; serum levels of sHLA-I were higher than the cutoff in ten patients with pancreatic cancer, and serum levels of CA19-9 were lower than 37 IU/l in 9 of 14 patients; sensitivity and specificity were 88.2% and 85.5%, respectively. Serum levels of sHLA-I in pancreatic cancer patients were higher than in the other diseases, although we found that pancreatic cancer cell lines did not produce the sHLA-I. The evaluation of serum sHLA-I levels could have clinical significance in pancreatic cancer. PMID- 11390037 TI - Typing for all known MICA alleles by group-specific PCR and SSOP. AB - Major histocompatibility complex class I chain-related gene (MICA) is a recently discovered polymorphic gene in the HLA region expressed mainly by certain epithelial cells, keratinocytes, endothelial cells, fibroblasts, and monocytes. MICA is structurally quite different from the HLA class I genes and is potentially associated with some diseases and with immune response to transplants. Some DNA-based typing techniques have previously been described for MICA including sequence-based typing (SBT) and analysis of single strand conformational polymorphisms (SSCP). In the present experiments we have developed a strategy that allows identification of all 54 MICA alleles described so far, using group-specific polymerase chain reactions (PCR) and sequence-specific oligonucleotide probes (SSOP). To analyze for the polymorphisms in exons 2, 3, and 4 an initial screening with group-specific primers, based on polymorphism at position 69 of exon 2, and at position 615-616 of exon 4, was used to determine four major groups of alleles. Then group-specific PCR amplifications were performed and the amplified DNA was hybridized with the corresponding panels of SSOP. An additional amplification was performed with locus-specific primers and hybridized with a set of SSOP to identify and/or confirm the presence of some of the alleles. Unequivocal MICA typing was achieved for 97 of 103 individuals. Of 54 previously described alleles, only 14 were observed in this population. One unexpected hybridization pattern was observed, and molecular cloning and sequencing confirmed it to be a novel sequence, which was given the local designation MICA-055D. The gene frequencies among 103 unrelated North American Caucasian donors were determined and the linkage disequilibrium between MICA and HLA-B was analyzed. PMID- 11390038 TI - Polymorphism in MICA rather than HLA-B/C genes is associated with psoriatic arthritis in the Jewish population. AB - The aim of this study was to examine whether the association of psoriatic arthritis (PsA) with human leukocyte antigen (HLA) class I genes is secondary to linkage disequilibrium with a nearby gene. We examined a sample of the Jewish population to investigate whether HLA-B/C and DR polymorphism is associated with susceptibility, or whether other closely related class I loci, such as the major histocompatibility complex class I chain-related gene A (MICA) and tumor necrosis factor (TNF), might play a role in disease development. Comparisons of different populations with different HLA profiles would be of value in identifying the candidate genes involved in PSA. Fifty-two patients with PsA and 73 random matched controls from a Jewish population were selected and DNA typed by polymerase chain reaction-single-strand oligonucleotide probe (PCR-SSOP) (HLA-C), PCR sequence-specific primers (PCR-SSP) (HLA-B, -DR), radioactive PCR (MICA-TM polymorphism in the transmembrane region), and PCR-RFLP (TNF). Some findings can be concluded from the study: (1) the frequency of HLA-B*5701, B*3801, B*39, B*27, Cw*0602, Cw*07, DRB1*0402, and DRB1*0701 were not found to be significantly increased in PsA; (2) no significant differences of TNFalpha promoter alleles at positions -308 and -238 were found between PsA and healthy controls; (3) the trinucleotide repeat polymorphism MICA-A9 was present at a higher frequency in PsA patients, (p(c) < 0.009, RR = 3.34, EF = 0.39); and (4) MICA-A9 polymorphism was found in linkage disequilibrium with HLA-B alleles (B*5701, B*3801) described to be associated with PsA in Caucasians. These results suggest that the MICA gene or other nearby gene(s) may be involved in the development of PsA, and it would thus appear that psoriasis vulgaris (PsV) and PsA are associated with different MHC susceptibility genes. PMID- 11390039 TI - Sequence-based typing of HLA class I alleles in Alaskan Yupik Eskimo. AB - In comparison to South America, native North Americans tend to be less diverse in their repertoire of HLA class I alleles. Based upon this observation, we hypothesized that the Yupik Eskimo would exhibit a limited number of previously identified class I HLA alleles. To test this hypothesis, sequence-based typing was performed at the HLA-A, -B and -C loci for 99 Central Yupik individuals from southwestern Alaska. Two new class I alleles, A*2423 and Cw*0806, were identified. While A*2423 was observed in only one sample, Cw*0806 was present in 26 of the 99 individuals and all of the Cw*0806 samples contained B*4801. Allele Cw*0806 differs from Cw*0803 by a single nucleotide substitution such that Cw*0803 may be the progenitor of Cw*0806. Allele Cw*0803 was originally characterized as unique to South America, but detection of Cw*0803 in the Yupik indicates that Cw*0803 was a founding allele of the Americas. The presence of new alleles and previously unrecognized founding alleles in the Yupik population show that natives of North America are more diverse than previously envisioned. PMID- 11390040 TI - Analysis of the distribution of HLA-B alleles in populations from five continents. AB - A two stage PCR-SSOP typing procedure, that permitted HLA-B allele assignment, was applied to DNA samples obtained from six diverse populations -Brazilian, Mexican (Series and Mestizos), Cuban (Caucasoid and Mulatto), South African Zulu, Omani, and Singapore Chinese. HLA-B allele frequencies and HLA-A/B two locus haplotype frequencies were compiled for each population. PMID- 11390041 TI - Human natural chimerism: an acquired character or a vestige of evolution? AB - Analysis on five common classes of human natural chimeras (cytomictical, whole body, fetal-maternal, germ cell, and tumor chimeras) reveals that (1) they initiate only during pregnancy, (2) the most common class are chimeras which contain maternal cells, and (3) the primary mechanisms that are involved in their formation and establishment are still elusive. These classes of natural chimerism, are involved only with maladaptive phenomena such as malignancy and autoimmune diseases and without any documented benefit. A recent review has challenged the accepted dogma that the evolution of immunity is pathogen-directed and asserted that preserving individuality from littering the soma and the germline by conspecific alien cells might have been the original function of the innate immunity. Following this tenet, I propose here that human natural chimerism is a by-product of the new role evolved from primitive components of immunity to "educate" the developing embryo with the armamentarium of effector mechanisms, dedicated to purge the individual from pervasive somatic and germline variants, and is not a vestige of evolution. PMID- 11390042 TI - Comparative ability of EMG, optimization, and hybrid modelling approaches to predict trunk muscle forces and lumbar spine loading during dynamic sagittal plane lifting. AB - OBJECTIVE: To compare the ability of three modelling approaches to resolve the muscle and joint forces in a lumbar spine model during dynamic sagittal plane lifting. DESIGN: Trunk muscle forces, spine compression, and coactivity predicted through double linear optimization, EMG-assisted, and EMG assisted by optimization approaches were compared.Background. The advantages of EMG-based approaches are known from static task analyses. Limited assessment has been made for dynamic lifting. METHODS: Eleven male subjects performed sagittal plane lifting-lowering at fixed cadence from 0 degrees to 45 degrees of trunk flexion with and without an external load of 12 kg. Three-dimensional kinematics and dynamics as well as surface EMG provided inputs to a 12 muscle lumbar spine model. RESULTS: Trunk muscle coactivity was different between the modelling approaches but spine compression was not. Both EMG-based approaches were sensitive to trunk muscle coactivity and imbalance in left-right muscle forces during sagittal plane lifting. Overall, the best correlations between predicted forces and EMG as well as between forces predicted by different modelling approaches were obtained with the EMG-based models. Only the EMG assisted by optimization approach simultaneously satisfied mechanical and physiological validity. CONCLUSIONS: Both EMG-based approaches demonstrated their potential to detect individual trunk muscle strategies. A more detailed trunk anatomy representation would improve the EMG-assisted approach and reduce the adjustment to muscle force gain through EMG assisted by optimization. RELEVANCE: Injury to the lumbar spine could command alternative strategies of motion to attenuate pain and damage. To understand these strategies, the ideal lumbar spine model should predict individual muscle force patterns and satisfy mechanical equilibrium. PMID- 11390043 TI - Tomodensitometry measurements for in vivo quantification of mechanical properties of scoliotic vertebrae. AB - OBJECTIVE: This in vivo study investigated the mechanical properties of scoliotic vertebrae especially in the apical zone. DESIGN: A method based on computed tomography images and finite element meshing had been developed to quantify and visualise the bone density distribution of scoliotic vertebrae. BACKGROUND: Most of scoliotic studies performed considered only geometrical parameters. METHOD: Computed tomography examination had been performed on 11 girls presenting idiopathic scoliosis. Using in-house image processing software and the pre-post processor Patran, a finite element mesh of each vertebral body and a mapping of each cancellous bone slice were proposed allowing the bone density distribution to be visualised. The mechanical properties were derived from predictive relationships between Young's modulus and computed tomography number. Geometrical (unit mass) and mechanical centres were calculated and compared in order to quantify the role of mechanical property distribution on the apex zone of the scoliotic spine. RESULTS: In the coronal plane, compared to the geometrical centre, the mechanical centre was shifted forward in the concavity (0.54 mm) of the curvature except for two vertebrae. In the sagittal plane, the mechanical centre was shifted forward in the back (0.26 mm) except for three vertebrae. The shift forward by slice was made in a same way for each slice (0.63 mm), except at the end plates (0.58 mm). DISCUSSION: The result values obtained were small but significant because the curvatures were low and the vertebrae were not wedged. Besides, one can observe that the scoliotic deformation evolution seemed to modify the mechanical property distribution. RELEVANCE: This study suggested the following question: Could these CT measurements be a predictive tool in scoliosis treatment? PMID- 11390044 TI - Cervical strength of young adults in sagittal, coronal, and intermediate planes. AB - OBJECTIVES: To measure the cervical isometric force generation capacity of men and women reliably while seated in upright neutral posture. DESIGN: The cervical muscle isometric strength was determined in flexion, extension, lateral flexion, anterolateral flexion and posterolateral extensions in an upright seated neutral posture. For this measurement a device was designed, fabricated and its reliability established. BACKGROUND: To-date only a few studies on cervical strength data have been published. Of these, validity of some data is suspected due to the use of unstandardized methodology. No studies were identified which reported cervical strengths in lateral and oblique planes. METHODS: A testing device consisting of sturdy, stable and strong telescopic adjustable square metal tube was firmly bolted in the floor. Another rotating metal tube was pivoted and adjustably counterweighted and attached to an immovable object with a load cell in its path. Using a horizontal bar upholstered at the terminal end attached to the rotating tube 40 young subjects were tested. They exerted their maximal voluntary isometric contraction in flexion, extension, lateral flexion, anterolateral flexion, and posterolateral extension bilaterally. RESULTS: Cervical muscle strength was maximum in extension and minimal in anterolateral flexion (which was very close to flexion strength). With progressive change in direction towards posterior region the strength progressively increased. There was a significant difference between male and female strengths (P < 0.01). The flexion/extension ratio of males was 1:1.37 and for females 1:1.79. There was a significant difference in strength values in different directions (P < 0.01). CONCLUSIONS: The cervical strength is direction dependent. The extension generates maximum force and flexion close to minimum. A progressive change from anterior to posterior direction generates increasing force. RELEVANCE: There is insufficient information regarding cervical strength, which may at least indirectly and in part, indicate the force bearing capacity of cervical musculature. The present study furnishes some data to this end. PMID- 11390045 TI - An in vivo technique for determining 3D muscular moment arms in different joint positions and during muscular activation - application to the supraspinatus. AB - OBJECTIVE: To develop a 3D in vivo technique for determining the moment arm and insertion angle of muscles in different joint positions and under muscular activity. DESIGN: An open magnetic resonance imaging system and 3D processing techniques were used for determining the moment arm and insertion angle of the supraspinatus in healthy volunteers. BACKGROUND: Muscular moment arms are important parameters for the computation of joint forces, however, so far in vivo measurements have not considered the influence of muscle activity and were restricted to two dimensions. METHODS: 10 healthy shoulders were investigated in 30-150 degrees abduction with and without abducting muscular activity, using an open magnetic resonance imaging. The minimal distance between the midpoint of the humeral head and the line of action of the supraspinatus was determined in 3D. The insertion angle was derived by calculating the angle between the humerus and the supraspinatus. RESULTS: During elevation a significant (P < 0.001) increase of the supraspinatus moment arm was observed (19.9 mm (SD, 2.3 mm) at 30 degrees; 23.3 mm (SD, 2.5 mm) at 150 degrees ), being significantly larger at 90 degrees and 120 degrees under muscle activity. The insertion angle also increased significantly during elevation (P < 0.001), the values becoming larger under muscle activity at 60 degrees (P < 0.01) and at 90 degrees (P < 0.05). CONCLUSIONS: This 3D technique permits to determine the moment arm and insertion angle of muscles in vivo not only in various joint positions but also under in vivo muscle activity. For the supraspinatus, we observed a constant increase of both parameters during arm abduction. RELEVANCE: The technique and these data can be used for improved computation of joint forces in biomechanical model, and for precise diagnostics in patients with altered scapulohumeral motion patterns. PMID- 11390046 TI - A numerical solution to calculate internal-external rotation at the glenohumeral joint. AB - OBJECTIVE: To validate the approach of using angular velocity vectors to quantify internal-external rotation of the humerus. DESIGN: An experimental approach was used to compare predictions of internal-external rotation of the humerus based on angular velocity vectors, and known measurements of internal-external rotation. BACKGROUND: A primary concern associated with description of glenohumeral biomechanics is measurement of internal-external rotation of the humerus. Euler angles are often used, yet they do not address problems such as Codman's paradox and 'gimbal lock'. Previous work has presented a technique that uses angular velocity vectors to quantify internal-external rotation instead of Euler angles. This approach is promising with regard to providing an independent measure of internal-external rotation of the humerus, and requires validation for subsequent use on humans. METHODS: A gimbal with three axes of rotation that simulated the rotational d.o.f. of the humerus relative to the glenoid was developed and used to validate the use of angular velocity vectors to quantify internal-external rotation of the humeral shaft portion of the gimbal. RESULTS: Correlations between calculated and measured rotation values revealed R(2) values of 0.99, slopes at diagonal (1.0), and y-intercepts near zero (-0.6 degrees ).Conclusions. The expression developed here is a valid and useful method for measuring motion of the humerus relative to the glenoid. RELEVANCE: Angular velocity vectors can be used to accurately determine internal-external rotation of the humerus relative to the glenoid, and this may be useful for the development of arthrometers to characterize the glenohumeral joint. PMID- 11390047 TI - The effect of forearm rotation on laxity and stability of the elbow. AB - OBJECTIVE: The purpose of this study was to quantify the relationship between forearm rotation and valgus/varus laxity of the elbow joint over the range of elbow flexion. BACKGROUND: There is little known about the influence of forearm rotation on the laxity and stability of the elbow joint. The general opinion exists that forearm rotation does not significantly influence the laxity and stability of the elbow joint. METHODS: Nine fresh-frozen cadaver elbows were used. Passive elbow flexion with the forearm in neutral rotation and in 40 degrees and 80 degrees of pronation and supination was performed under valgus/varus loads: (1) in intact elbows; (2) after a lateral surgical approach (lateral epicondylar osteotomy of the distal humerus); (3) after release of the anterior bundle of the medial collateral ligament; and (4) after release of the anterior bundle of the medial collateral ligament plus radial head resection. Valgus/varus elbow laxity was quantified using an electromagnetic tracking device. RESULTS: There was a statistically significant effect (P < 0.05) of forearm rotation on valgus/varus laxity throughout the range of flexion. The laxity was always greater in pronation than in supination, regardless of the surgical approach or the integrity of the anterior bundle of the medial collateral ligament or radial head. CONCLUSIONS: Valgus/varus laxity of the elbow is forearm rotation-dependent. The potential role of this effect should be considered and controlled for in the design of studies examining laxity and stability of the elbow joint. RELEVANCE: The observation that forearm pronation increases valgus/varus laxity, particularly in medial collateral ligament deficient elbows, implies a possible additional factor in throwing kinematics that might put professional baseball pitchers at risk of medial collateral ligament injury due to chronic valgus overload. Our data indicate that forearm rotation should be considered during the clinical examination of elbow instability. PMID- 11390048 TI - Elbow load during pushup at various forearm rotations. AB - OBJECTIVE: Elbow joint loading was evaluated during pushup exercises at various forearm rotations. DESIGN: Subjects were asked to perform pushup in various forearm rotations: neutral, 90 degrees internal rotation, and 90 degrees external rotation. BACKGROUND: Training with pushup exercise is good for the muscles and joints of the upper extremities. However, excessive shear forces on the elbow might lead to injuries to either normal trainees or to handicapped people, especially for those who rely on elbow prosthesis. METHODS: The kinematics and kinetics of the elbow joint were investigated under various forearm rotations. RESULTS: The loading biomechanics of the elbow joint differed with various forearm rotations. It was noted that greater posterior and varus forces of the elbow are encountered with internal rotation of the hand position and, consequently, full forearm pronation. CONCLUSIONS: Pushup with hands in internally rotated position should be prevented so as to avoid excessive shear forces or moments. RELEVANCE: Knowledge of elbow kinematics and kinetics may be helpful in preventing injuries by reducing the elbow shear force with changes of forearm rotation. PMID- 11390049 TI - A computer model to simulate patellar biomechanics following total knee replacement: the effects of femoral component alignment. AB - OBJECTIVE: The objective of this study is to analyze the biomechanics of the patellar component following total knee replacement. More specifically we investigated the effect of displacing the femoral component of an Insall-Burstein II total knee replacement on the patellar tracking and patello-femoral contact pressures. DESIGN: We used a validated computer simulation of the knee joint to virtually insert the femoral component with the following four types of placements: (1) no misplacement, (2) 5 degrees of internal rotation, (3) 5 degrees of external rotation and (4) 5 degrees of flexion rotation. The patellar 3D tracking and patello-femoral contact pressures were computed for each femoral component placement as a function of knee flexion angle. BACKGROUND: Complications at the patello-femoral joint are the among most frequent following total knee replacement. RESULTS: Femoral component placement unevenly affected the associated patellar tracking: a 5 degrees internal rotation tilted and rotated the patella laterally by about 5 degrees throughout knee flexion. A 5 degrees external rotation of the femoral component had less effect on patellar tracking. A rotation of 5 degrees in flexion primarily caused patellar rotation (5-10 degrees lateral rotation). Femoral component malalignment had only minor effects on the peak pressure distributions at the patello-femoral interface. CONCLUSION: These results suggest that femoral component positioning primarily affects patellar tracking, with a possible threat for patellar subluxation under external rotation of the femoral component. RELEVANCE: Precise alignment of the prosthetic components is difficult to control during total knee replacement due to the lack of precise anatomical landmarks in the human knee joint. Consequently, the position of each prosthetic component may differ from the ideal one suggested by the manufacturer. Improper alignment of the prosthetic components during total knee replacement may lead to premature implant failure. PMID- 11390050 TI - Patellofemoral joint kinetics during squatting in collegiate women athletes. AB - OBJECTIVE: To characterize the biomechanics of the patellofemoral joint during squatting in collegiate women athletes. DESIGN: Repeated measures experimental design. BACKGROUND: Although squatting exercises are required components of most intercollegiate resistance-training programs and are commonly performed during rehabilitation, the effects of various squatting depths on patellofemoral joint stress have not been quantified. METHODS: Anthropometric data, three-dimensional knee kinematics, and ground reaction forces were used to calculate the knee extensor moment (inverse dynamics approach) in five intercollegiate female athletes during squatting exercise at three different depths (approximately 70 degrees, 90 degrees and 110 degrees of knee flexion). A biomechanical model of the patellofemoral joint was used to quantify the patellofemoral joint reaction force and patellofemoral joint stress during each trial. RESULTS: Peak knee extensor moment, patellofemoral joint reaction force and patellofemoral joint stress did not vary significantly between the three squatting trials. CONCLUSIONS: Squatting from 70 degrees to 110 degrees of knee flexion had little effect on patellofemoral joint kinetics. The relative constancy of the patellofemoral joint reaction force and joint stress appeared to be related to a consistent knee extensor moment produced across the three squatting depths. RELEVANCE: The results of this study do not support the premise that squatting to 110 degrees places greater stress on the patellofemoral joint than squatting to 70 degrees. These findings may have implications with respect to the safe design of athletic training regimens and rehabilitation programs. PMID- 11390051 TI - The influence of ergometer pedaling direction on peak patellofemoral joint forces. AB - OBJECTIVE: To compare patellofemoral joint forces during forward pedaling and reverse pedaling on a bicycle ergometer. DESIGN: Experimental within-subject design. BACKGROUND: Clinicians addressing patellofemoral pain syndrome often strive to achieve quadriceps strength while reducing forces at the patellofemoral joint. Related to this goal, previous research has shown greater activity in the quadriceps during reverse pedaling compared to forward pedaling on a bicycle ergometer, however, patellofemoral joint forces were not examined. METHODS: Twenty-one healthy males performed 5-min exercise bouts for forward and reverse pedaling while normal and tangential pedal forces and kinematic data were collected. Inertial properties of each segment were estimated and net joint moments were calculated using Newtonian inverse dynamics. Patellofemoral joint forces (Fpf) were quantified using the equation Fpf = 2FQ . sin beta/2, where FQ is the quadriceps force and beta is the patellar mechanism angle. RESULTS: Reverse pedaling exhibited a 110% greater peak patellofemoral joint force and a 149% greater quadriceps force than forward pedaling. Peak effective moment arm lengths were not different between conditions. CONCLUSIONS: Reverse pedaling is associated with a greater peak patellofemoral force than forward pedaling. The peak force was influenced by the quadriceps force and not by the moment arm length. RELEVANCE: Insufficient data exist concerning the use of reverse pedaling as an alternative to forward pedaling for rehabilitation of knee disorders. Although quadriceps activity appears greater for reverse pedaling compared to forward pedaling, greater peak patellofemoral forces contradict clinical treatment goals for patellofemoral pain syndrome and may lead to load-elicited pain or damage to joint structures. PMID- 11390052 TI - A comparison of knee joint motion patterns between men and women in selected athletic tasks. AB - BACKGROUND: Women have higher non-contact anterior cruciate ligament injury rate than men do in sport activities. Non-contact anterior cruciate ligament injuries frequently occur in sports requiring cutting tasks. Alternated motor control strategies have identified as a potential risk factor for the non-contact anterior cruciate ligament injuries. The purpose of this study was to compare the patterns of knee kinematics and electromyographic activities in running, side cutting, and cross-cutting between men and women recreational athletes. METHODS: Three-dimensional kinematic data of the knee and electromyographic data of selected muscles across the knee joint were collected for 11 men and 9 women recreational athletes in running, side-cutting, and cross-cutting. Regression analyses with dummy variables for comparison of knee motion patterns between men and women. RESULTS: Women tend to have less knee flexion angles, more knee valgus angles, greater quadriceps activation, and lower hamstring activation in comparison to men during the stance phase of each of the three athletic tasks. Literatures suggest these alternated knee motion patterns of women tend to increase the load on the anterior cruciate ligament. CONCLUSION: Women on average may have certain motor control strategies that may alter their knee motion patterns. Women's altered knee motion patterns may tend to increase the load on the anterior cruciate ligament in the selected athletic tasks, which may contribute to the increased anterior cruciate ligament injury rate among women. RELEVANCE: Non-contact anterior cruciate ligament injuries frequently occur in sports. Altered motor control strategies and lower extremity motion patterns are likely to play an important role in non-contact anterior cruciate ligament injuries. Non-contact anterior cruciate ligament injuries may be prevented by correcting altered motor control strategies and associated lower extremity motion patterns through certain training programs. PMID- 11390053 TI - Pattern of abnormal tangential forces in the diabetic neuropathic foot. AB - OBJECTIVES: The role of tangential stress in neuropathic foot ulceration is yet unknown. The aim of this study was to investigate the tangential forces developed during gait by the whole foot and by selected subareas of it, namely the heel, the metatarsals and the hallux. METHODS: 61 diabetic patients have been evaluated: 27 without neuropathy, 19 with neuropathy and 15 with previous neuropathic ulcer. The patients were compared with 21 healthy volunteers. A piezo dynamometric platform was used to measure the three components of the ground reaction force under the total foot and the selected subareas. RESULTS: A significant reduction was observed for the forward peak and the backward peak of the anteroposterior ground reaction force component measured under the whole foot. Patients with previous neuropathic ulcer showed a significant increase of the mediolateral stress under the metatarsals. CONCLUSIONS: Tangential stress is altered in diabetic neuropathic patients; the increased mediolateral component suggests that tangential stress could have a role in the high risk of recurrence observed in patients with previous ulceration. RELEVANCE: To assess the effectiveness of a non-invasive methodology for the estimation and the monitoring of significant alterations of the tangential stress with the increase of neuropathy. PMID- 11390069 TI - Standardised tests in mice and cattle for the detection of drug resistance in tsetse-transmitted trypanosomes of African domestic cattle. AB - Resistance to the drugs used to control African animal trypanosomosis is increasingly recognised as a constraint to livestock production in sub-Saharan Africa. The most commonly used tests for detection of trypanocidal drug resistance are tests using mice or ruminants, but these suffer from lack of standardisation and hence it may be difficult to compare the results of different investigators. Tests in mice are less expensive than tests in ruminants, but while tests in mice they may be useful as a general guide to resistance in a geographic area they should not be extrapolated to cattle on an individual trypanosome level. Moreover, the commonly used protocols are too laborious for their application to large number of trypanosome isolates on an area-wide basis. This paper presents guidelines for standardised testing of trypanocidal drugs in vivo, and introduces a simplified single-dose test for use in mice, which is convenient for use in areas with limited laboratory facilities. The single-dose test is appropriate for characterisation of geographic areas in terms of trypanocidal drug resistance using large numbers of trypanosome isolates, for making comparisons between areas, and for monitoring changes in trypanocidal drug resistance over time. Multiple-dose tests may be used to determine the degree of resistance of individual stabilates to be determined precisely in mice are also described, but for logistical reasons these will rarely be conducted on more than a few stabilates, and testing of a larger number of stabilates in the single-dose test will generally provide more useful information. Finally, we describe tests in cattle that may be used to determine the efficacy of recommended curative doses of trypanocidal drugs for the treatment of infection with individual trypanosome isolates, including Trypanosoma vivax, which is rarely infective for mice. PMID- 11390070 TI - General expression of RoTat 1.2 variable antigen type in Trypanosoma evansi isolates from different origin. AB - The variable surface glycoprotein of Trypanosoma evansi RoTat 1.2 variable antigen type (VAT) is used as an antigen in different antibody detection assays for T. evansi. To obtain more information on the predominant character of RoTat 1.2 and its diagnostic potential in antibody detection tests, we checked its expression in 10 different T. evansi stocks and clones from different parts of the world. Cryostabilates were injected into mice and the trypanosomes of the first peak parasitaemia were screened for the presence of RoTat 1.2 by VAT specific immunofluorescence. To monitor the appearance of RoTat 1.2 specific antibodies during infection, rabbits were infected and serologically tested at different time intervals with VAT specific immune trypanolysis, CATT/T. evansi, LATEX/T. evansi and ELISA/T. evansi. Test results confirm the predominant character of RoTat 1.2. PMID- 11390071 TI - The detection of Toxoplasma gondii by comparing cytology, histopathology, bioassay in mice, and the polymerase chain reaction (PCR). AB - The objective of this study was to compare the different methods of detecting Toxoplasma gondii in sheep tissue, tested serologically positive by the indirect immunofluorescent antibody test (IFAT). Brain, diaphragm, and blood samples were collected from 522 sheep slaughtered at the Sao Manuel abattoir, Sao Paulo State, Brazil. Brain and diaphragm samples from IFAT seropositive animals were digested by both trypsin and pepsin and then injected into mice. Part of the digested samples was used to prepare slides for Giemsa staining and in the polymerase chain reaction (PCR). Tissue fragments were fixed in formalin and examined using hematoxilin-eosin (HE). Forty of the sheep (7.7%) were IFAT positive. T. gondii was isolated in 23 (59.0%) of the 39 mice with pepsin-digested brain samples and in 27 (69.0%) of the 39 with trypsin-digested brain samples. Injection of diaphragm samples led to T. gondii isolation in 26 (66.7%) of the 39 pepsin digested samples and 21 (53.8%) of the 39 trypsin-digested samples. Cytological and hystopathological examination of both brains and diaphragms was negative in all examined sheep. PCR was positive in 7 (17.9%) of the trypsin and 2 (5.1%) of the pepsin-digested samples, while 9 (23.1%) of the trypsin and 3 (7.7%) of the pepsin-digested samples showed T. gondii DNA. T. gondii isolation rate in mice (n = 34; 85.0%) was significantly higher than detection by PCR (n = 15; 37.5%). PMID- 11390072 TI - A multiplex PCR assay for differentiating economically important gastrointestinal nematodes of cattle. AB - A multiplex polymerase chain reaction (PCR) test was developed for identifying gastrointestinal (GI) nematodes that commonly infect cattle. This assay was developed using adult-derived genomic DNA and shown capable of discriminating parasite eggs from the feces of experimentally-infected animals at both the species and genus levels. Sequence data from internal (ITS) and external (ETS) transcribed spacers of the ribosomal DNA (rDNA) repeats as well as the 3'-end of the small subunit rDNA and 5'-end of the large subunit rDNA were used to generate five primer sets which, when used simultaneously in a multiplex PCR, produce a unique electrophoretic DNA banding pattern characterized by a single DNA fragment for Ostertagia ostertagi (257bp), Haemonchus placei (176bp), Oesophagostomum radiatum (329bp), Trichostrongylus colubriformis (243bp) and Cooperia oncophora (151bp). In a similar manner, the constructed primer sets amplified DNA from Ostertagia lyrata, Haemonchus contortus, Trichostrongylus axei, Cooperia surnabada and Cooperia punctata. With respect to H. contortus, a closely migrating doublet was generated suggesting size heterogeneity in the ETS which is consistent with multiple rDNA repeat units within this species. PCR analyses using mixtures of monospecifically-purified nematode eggs indicated a sensitivity of less than 0.5 egg-DNA equivalent per species. Although, not designed as a quantitative technique, relative PCR signal intensities corresponded to relative egg burdens within the DNA samples from mixed species of eggs. PMID- 11390073 TI - Resistance of Cooperia to ivermectin treatments in grazing cattle of the Humid Pampa, Argentina. AB - A field experiment and a controlled efficacy trial were conducted to demonstrate resistance of cattle trichostrongylid nematodes to endectocidal compounds in grazing cattle on the Humid Pampa, Argentina. Male Polled Hereford calves, aged 9 11 months old, with a history of frequent treatments with compounds of the avermectin/milbemycin class, were used for the trial. The field experiment involved six groups of 10 animals each, which were subcutaneously treated with either ivermectin (IVM), long-acting ivermectin (LA-IVM), moxidectin (MXD) or doramectin (DRM) at a dosage of 0.2mg/kg BW. A fifth group was treated orally with fenbendazole at a dosage of 5mg/kg BW; the sixth group of calves served as non-treated control. Faecal samples were collected from each animal on the day of treatment and at 14 days after treatment. Nematode egg counts were performed to estimate the faecal egg count reduction test (FECRT). The FECRT showed that reductions were 65% (IVM), -20% (LA-IVM) and 85% (DRM). Egg counts from calves treated with MXD or fenbendazole were reduced by 95 and 100%, respectively. For the controlled efficacy trial, six animals with the highest egg counts from the IVM, LA-IVM and non-treated control groups, were necropsied on day 18 after treatment to determine numbers and nematode species in the abomasum and small intestine. The results indicated that efficacy against Cooperia oncophora was 62.7 and 48% for IVM and LA-IVM, respectively. Neither of the IVM formulations showed efficacy against Trichostrongylus spp. in the small intestine. However, the absence of Trichostrongylus spp. in some animals of both treated and control groups precluded a valid assessment of efficacy or resistance. It was concluded that resistance of trichostrongylid nematodes to the avermectin/milbemycin class of compounds in grazing cattle of the Humid Pampa, Argentina, may be strongly established on farms where cattle are treated at frequent intervals throughout the year. PMID- 11390074 TI - Control of anthelmintic resistant endoparasites in a commercial sheep flock through parasite community replacement. AB - An effort was undertaken to replace a community of sheep endoparasites that had been classified as resistant to levamisole and albendazole with a community of more susceptible parasites using a dilution approach that could be integrated into the management of a commercial flock. For this study, pastures on this sheep farm were divided into two areas: north and south. Strategically timed anthelmintic treatments combined with pasture management reduced to nondetectable levels the endemic community of anthelmintic resistant parasites in this flock and on these pastures by early summer. A group of 102 ewes, lambs, and rams were experimentally infected with third stage larvae from the more susceptible community of parasites. These sheep then seeded the south pastures with the new parasite community, while sheep on the north pastures maintained the endemic resistant community. Despite its insensitivity as a technique for detecting anthelmintic resistance, fecal egg count reduction tests at the end of the grazing season indicated that the more susceptible parasites were present on the south pastures while resistant parasites were present on the north. The following grazing season, similar protocols were used to introduce the more susceptible parasites onto the north pastures. At the end of the grazing season, fecal egg count reduction tests indicated that the new community of parasites had become established on both groups of pastures of the farm. PMID- 11390075 TI - Development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle. AB - The present study reports on the development of a copro-antigen capture ELISA for detecting Ostertagia ostertagi infections in cattle. The ELISA was based on polyclonal rabbit antibodies, which recognize O. ostertagi excretory/secretory antigens (ES). ES antigens are released by the metabolic active stages of the parasite in the abomasum, and passed in the faeces of the host. The detection limit of pure ES material was 30 ng ml(-1) in sample buffer and 125 ng ml(-1) in faecal extract. The test was evaluated using a follow up from six artificially infected calves. Elevated levels of Ostertagia coproantigens could be measured from 21 days after infection, indicating that only the presence of adult parasites can be detected. To evaluate the capacity of the assay to measure levels of infection, three groups of cattle were tested: 38 artificially infected calves, 17 naturally infected first grazing season calves and 16 naturally infected adult dairy cows. Optical densities were significantly correlated to the worm burdens of the animals and the ELISA had an overall sensitivity of 91% and a specificity of 45%. The test gave negative readings for faeces of animals carrying patent mono-infections with Cooperia oncophora. PMID- 11390076 TI - Prevalence of sparganosis by county of origin in Florida feral swine. AB - Sparganosis is a parasitic infection in amphibians, reptiles and mammals including feral swine and man. It is caused by migration of the metacestode (spargana) of Spirometra. The primary objective of this study was the determination of the prevalence of gross sparganosis in Florida county of origin in slaughtered feral swine. Tracebacks to county of origin were conducted for Florida feral swine with and without gross sparganosis. Feral swine trapped in Florida and presented for slaughter in a Texas slaughter establishment from May to December 1999 was the sample population. Overall prevalence of sparganosis in Florida feral swine was 6.9%. Because Highlands county had the same prevalence, other counties were compared to it. Sparganosis was detected in 17 Florida counties. Swine originating from Osceola or Hillsborough counties (4.3 and 1.8% prevalence, respectively) had lower prevalence of sparganosis than in Highlands, whereas those from Marion county (21.7% prevalence) had a higher prevalence. Transmission to humans may occur via consumption of infected feral swine, other species of secondary intermediate hosts or the primary intermediate hosts. PMID- 11390077 TI - Canine ocular onchocercosis in Hungary. AB - An adult male mongrel dog that had spent its entire life in Hungary, was found to have infection with filaroid nematodes of the genus Onchocerca. The gravid male and female parasites were embedded in bean-sized granulomatous masses on the conjunctiva and the sclera of both eyes. The cuticle of females consisted of two separated layers in longitudinal sections, the external layer bearing ridges and the internal layer showing striations. The ridges were marked, rounded in shape, and the ratio of body diameter to the distance between ridges varied between 7:1 and 10:1. At midbody of the worms, two striations could be seen between each pair of ridges: one under every ridge and one between neighbouring ridges. Numerous exceptionally small (96.4 microm x 6.4 microm) microfilariae were seen in the uteri of females and the surrounding tissues and isolated from skin biopsy materials. The morphology and location of the parasite and histopathological lesions of the Hungarian case were similar to that described in dogs in the United States. This case is the first documented ocular Onchocerca infection in dogs outside the western United States. Thus, onchocercosis should be considered in the differential diagnosis of ocular and periocular nodules in dogs also in Europe. PMID- 11390078 TI - Drought and flock isolation may enhance the development of anthelmintic resistance in nematodes. AB - A survey of anthelmintic resistant nematodes was conducted in sheep and goat flocks in Greece using in vivo and in vitro tests. Faecal egg count reduction tests in Macedonia were all greater than 99% indicating very high sensitivity of the nematodes to anthelmintics. In vitro tests showed benzimidazole resistant Teladorsagia circumcincta in 17 out of 106 flocks on small islands. On the mainland there were only three cases of benzimidazole resistance out of 310 flocks and animals had recently been introduced to the flocks. Flocks on the islands are isolated and there are higher temperatures than on the more mountainous mainland, where flocks tend to intermingle. It is concluded that drought and isolation are likely to be the major factors accounting for the development of anthelmintic resistance in nematodes in the island flocks. PMID- 11390079 TI - Anthelmintic resistance on goat farms in Georgia: efficacy of anthelmintics against gastrointestinal nematodes in two selected goat herds. AB - Gastrointestinal nematode (GIN) parasitism is a major constraint to production of goats in the southeastern United States. The conventional method of control used by producers in this region is frequent use of anthelmintics during the warm season. Overuse of anthelmintics has led to an increase in the incidence of anthelmintic resistance in many parts of the world, but data on prevalence of anthelmintic resistance in GIN of goats in the southeastern United States are very limited. To address this issue, anthelmintic efficacy was determined in goat herds at the Fort Valley State University, Agricultural Research Station (FVSU ARS) and the University of Georgia, College of Veterinary Medicine (UGA-CVM) using fecal egg count reduction (FECR) tests and DrenchRite((R)) larval development assays (LDA). At FVSU-ARS, 2-year-old Spanish goat does were randomly allocated to one of nine different treatment groups (n = 10): albendazole (ABZ; 20mg/kg body weight (BW)), fenbendazole (FBZ; 20mg/kg BW), ivermectin (IVM; 0.4 mg/kg BW), doramectin (DRM; 0.4 mg/kg BW), moxidectin (MOX; 0.4 mg/kg BW), levamisole (LEV; 12 mg/kg BW), morantel tartrate (MOR; 10mg/kg BW), a combination of IVM (0.4 mg/kg BW) and ABZ (20 mg/kg BW), and untreated controls. At UGA-CVM, goats were randomly allocated to one of five different treatment groups (n = 8): ABZ (20 mg/kg BW), IVM (0.4 mg/kg BW), MOX (0.4 mg/kg BW), LEV (12 mg/kg BW), and untreated controls. All drugs in both experiments were administered orally. Anthelmintic efficacy was calculated by comparing 14-day post-treatment FEC of treated and control animals, and percent reductions were interpreted using the World Association for the Advancement of Veterinary Parasitology guidelines for resistance. For the LDA, nematode eggs were isolated from pooled fecal samples of untreated control goats in each herd and used to perform DrenchRite((R)) assays. In the FVSU-ARS herd, MOX, LEV, the combination of IVM and ABZ, IVM, DRM, ABZ, MOR, and FBZ reduced FEC by 100, 91, 88, 78, 76, 62, 48, and 10%, respectively. In the UGA-CVM herd, MOX, LEV, ABZ and IVM, reduced FEC by 100, 94, 87, and 0%, respectively. In both herds moxidectin was the only drug tested that was fully effective. Results of the LDA were in agreement with results of the FECR tests for both herds. These data demonstrate the presence of GINs resistant to all three major anthelmintic classes in both goat herds. PMID- 11390080 TI - A single pasture limited treatment approach to estimate production loss from gastrointestinal nematodes in grazing stocker cattle. AB - The objective of this study was to evaluate the use of a single pasture, limited treatment methodology to assess the impact of gastrointestinal nematodes on weight gain in grazing cattle. From a group of 450 British crossbred, yearling spayed heifers, 60 animals were randomly selected (prospective randomization) prior to placement on summer pasture. Each of these 60 animals was weighed, a fecal sample obtained for nematode egg count and a uniquely numbered ear tag applied. A randomly assigned group of 30 received treatment with one ivermectin sustained release bolus, while the remaining 30 served as non-treated controls. The treatment and control groups rejoined the remaining non-treated 390 animals, and were grazed as a single group for 165 days. At grazing conclusion, treatment and control cattle were individually weighed, and fecal samples obtained for nematode egg counts. Treatment was associated with a 0.064 kg per grazing day gain increase, or a 10 kg increase over the grazing season (P = 0.02). Nematode egg counts at grazing initiation were not different between treatment and control (P = 0.30), though egg counts in treated cattle at study end were lower than control (P < 0.0001). Results of this study support the use of a single pasture limited treatment approach for measuring the effect that internal nematodes have on weight gain in grazing cattle under commercial range conditions. PMID- 11390081 TI - Efficacy of a long-acting formulation of ivermectin against Psoroptes ovis (Hering, 1838) on cattle. AB - A study was conducted in cattle experimentally infested with Psoroptes ovis to compare the prophylactic control against P. ovis provided by a long-acting injectable formulation of ivermectin to that of a commercially available injectable formulation of doramectin. Thirty Holstein steers were used. Animals were allocated by restricted randomization based on Day 0 body weight, forming six replicates of five animals each. Within each replicate, one animal was randomly allocated to one of the following treatment groups, with ivermectin and doramectin administered subcutaneously where indicated: (1) untreated controls; (2) ivermectin long-acting injectable (LAI) 630 mcg/kg, 56 days before challenge; (3) ivermectin LAI 630 mcg/kg, 42 days before challenge; (4) ivermectin LAI 630 mcg/kg, 35 days before challenge; or (5) doramectin 200 mcg/kg, 35 days before challenge. Animals were housed in individual pens 1 week prior to treatment. All animals were experimentally infested with P. ovis mites in the area between the shoulders, on the same day. Live mites were counted in scrapings from mange lesions at 2 sites on each animal 14, 21 and 28 days after challenge. Live mites were found in 33, 67 and 83% of the untreated controls on each respective evaluation. No P. ovis mites were found in steers treated with ivermectin LAI. Those animals showed lower (P < 0.05) mite counts than untreated controls on evaluations conducted 21 and 28 days after challenge. These results indicate that the ivermectin long-acting injectable formulation prevents induced infestations by P. ovis for at least 56 days after treatment. Doramectin injectable formulation, used at 200 mcg/kg, did not have a prophylactic effect 35 days after treatment. PMID- 11390082 TI - Worm control practices of pig farmers in Greece. AB - Information concerning worm control practices of pig farmers in Greece was collected through a questionnaire survey by visiting farms and interviewing farmers. Questionnaires from 93 farmers were collected. The most commonly treated age groups of pigs were sows (87%), followed by fatteners (59%) and weaners (57%). Most of the farmers treated their sows twice per year (78%). The preferred method for calculating anthelmintic dosages was visual estimation (77%) of an average body weight (97%). Drug administration was by mixing anthelmintics with 1 day ration (97%) in two doses with an interval of 4-5 days (94%). The most commonly used group of anthelmintic was benzimidazoles and probenzimidazoles (37 46%), followed by avermectins (7-14%) and tetrahydropyrimidines (8-14%). Febantel was the most commonly used individual drug (34-40%), followed by ivermectin (7 14%) and pyrantel (9-14%). A small number of pig farmers used more than one group of anthelmintics (1-4%) or individual drug (2-6%) in the same year. The majority of the farmers (60%) used the same anthelmintic group for 3 or more years. When gilts/sows or boars were brought, 55% of the pig farmers did not treat with an anthelmintic. Most of the farmers evaluated the efficacy of an anthelmintic by inspecting the general appearance of the pigs or by inspecting for the presence of worms expelled in the faeces after treatment. Planning of the anthelmintic treatment strategy for most of the farms was the responsibility of veterinarians. However, most of the farmers considered the magnitude of worm infection in their own farm as insignificant. PMID- 11390083 TI - Reduction in mortality from heartwater in cattle, sheep and goats exposed to field challenge using an inactivated vaccine. AB - Inactivated vaccines for heartwater prepared with the commercially acceptable Montanide ISA 50 (ISA 50) adjuvant were field tested in Boer goats in Botswana, Angora goats in South Africa, and Merino sheep in Zambia and Zimbabwe. Two vaccines, one made using the Zimbabwean Mbizi isolate and the other using the respective local field isolate (Sunnyside in Botswana; Bathurst in South Africa; Lutale in Zambia), were tested at each site, except in Zimbabwe where only the Mbizi vaccine was tested. Compared with unvaccinated animals, the Mbizi vaccine significantly protected goats and sheep against field Amblyomma tick challenge in Botswana, Zambia and Zimbabwe (P = 0.018, 0.002 and 0.017, respectively), but failed to protect Angora goats in South Africa. However, in South Africa the vaccine prepared using the local field isolate Bathurst, induced significant protection (P=0.008). The vaccines containing the local isolates at all other sites were less protective than the Mbizi vaccine. The Mbizi inactivated vaccine also significantly protected 17 of 21 cattle (P = 0.05) against heartwater challenge from field ticks in Zimbabwe. Against the same challenge only 7 of 21 unvaccinated control cattle survived. This study demonstrates that heartwater is a major constraint to upgrading livestock in endemic areas, and caused an overall mortality of 77.6% in naive sheep and goats (97 of 125 died) and 67% in cattle (14 of 21 died). In contrast, the vaccine had a protective effect by reducing the overall mortality in sheep and goats to 54.3% (113 of 208 died) and to 19% in cattle (4 of 21 died). PMID- 11390084 TI - Current prevalence of adult Uncinaria spp. in northern fur seal (Callorhinus ursinus) and California sea lion (Zalophus californianus) pups on San Miguel Island, California, with notes on the biology of these hookworms. AB - A prevalence survey for hookworms (Uncinaria spp.) was done in northern fur seal (Callorhinus ursinus) and California sea lion (Zalophus californianus) pups on San Miguel Island, CA, in 2000. Intestines of dead pups were examined for adult hookworms in July. These parasites were found in 95% of 20 fur seal pups and 100% of 31 sea lion pups. The number of hookworms varied from 4 to 2142 (mean = 760) in fur seal pups and from 20 to 2634 (mean = 612) in sea lion pups. A direct relationship was evident between body condition and number of hookworms in the pups; that is, pups in poor condition had fewer hookworms than those in good condition. There was a decline in the number of hookworms in sea lion pups in 2000 compared to collections in 1996. Eggs of Uncinaria spp. were found in rectal feces (collected in late September and early October) of none of 35 (0%) live fur seal pups and 41 of 48 (85%) live sea lion pups. Packed cell volume values, determined for most of the same live pups, were essentially normal for C. ursinus but were much lower than normal for most Z. californianus. Hookworm larvae were not found in blubber of fur seal and sea lion pups or in rookery sand in July. Rookery sand, positive for live hookworm larvae when put in a refrigerator, was negative at removal 2.5 years later. The average number of eggs in utero of female hookworms was 285 for three specimens from a fur seal pup and 281 from three specimens from a sea lion pup. One hookworm larva was recovered from milk stripped from the teats of a stranded Z. californianus female at The Marine Mammal Center, Sausalito, CA. PMID- 11390085 TI - Dual Sarcocystis neurona and Toxoplasma gondii infection in a Northern sea otter from Washington state, USA. AB - Dual Sarcocystis neurona and Toxoplasma gondii infection was observed in a Northern sea otter from Washington, USA. The animal was found stranded, convulsed, and died shortly thereafter. Encephalitis caused by both S. neurona and T. gondii was demonstrated in histological sections of brain. Immunohistochemical examination of sections with S. neurona specific antisera demonstrated developmental stages that divided by endopolygeny and produced numerous merozoites. PCR of brain tissue from the sea otter using primer pairs JNB33/JNB54 resulted in amplification of a 1100 bp product. This PCR product was cut in to 884 and 216 bp products by Dra I but was not cut by Hinf I indicating that it was S. neurona [J. Parasitol. 85 (1999) 221]. No PCR product was detected in the brain of a sea otter which had no lesions of encephalitis. Examination of brain sections using T. gondii specific antisera demonstrated tachyzoites and tissue cysts of T. gondii. The lesions induced by T. gondii suggested that the sea otter was suffering from reactivated toxoplasmosis. T. gondii was isolated in mice inoculated with brain tissue. A cat that was fed infected mouse brain tissue excreted T. gondii oocysts which were infective for mice. This is apparently the first report of dual S. neurona and T. gondii in a marine mammal. PMID- 11390086 TI - Cloning and structural analysis of partial acetylcholine receptor subunit genes from the parasitic nematode Teladorsagia circumcincta. AB - Nematode nicotinic acetylcholine receptors (nAChRs) are the sites of action for the anthelmintic drug levamisole. Recent findings indicate that the molecular mechanism of levamisole resistance may involve changes in the number and/or functions of target nAChRs. Accordingly, we have used an RT-PCR approach to isolate and characterise partial cDNA clones (tca-1 and tca-2) encoding putative nAChR subunits from the economically important trichostrongyloid, Teladorsagia circumcincta. The predicted tca-1 gene product is a 248 aa fragment (TCA-1) which contains structural motifs typical of ligand-binding (alpha-) subunits, and which shows very high sequence similarities (98.8 and 97.2% amino acid identities) to the alpha-subunits encoded by tar-1 and hca-1 from Trichostrongylus colubriformis and Haemonchus contortus, respectively. Sequence analyses of partial tca-1 cDNAs from one levamisole-resistant and two susceptible populations of T. circumcincta revealed polymorphism at the predicted amino acid level, but there was no apparent association of any particular tca-1 allele with resistance. tca-2 encodes a 67 aa fragment (TCA-2) containing the TM4 transmembrane domain and carboxyl terminus of a putative nAChR structural (non-alpha) subunit. The deduced amino acid sequence of TCA-2 shows highest similarity (75% amino acid identity) to ACR-2, a structural subunit involved in forming levamisole-gated ion channels in Caenorhabditis elegans, but low similarity (43% identity) to the corresponding regions of TAR-1 and HCA-1. tca-2 is the first nAChR subunit gene of this type to be isolated from parasitic nematodes, and it provides a basis for further characterisation of structural subunits in trichostrongyloids. PMID- 11390087 TI - The rally call recognition in males of two hybridizing partridge species, red legged (Alectoris rufa) and rock (A. graeca) partridges. AB - The red-legged (Alectoris rufa) and rock (A. graeca) partridges hybridize and produce fertile offspring along a contact zone in French Southern Alps. The rally call emitted during pair formation, could play an important role in species recognition, acting as a behavioral reproductive isolating mechanism between males and females. In the present study, the coding system of the rally call was investigated from captive males of the two species and from F1 hybrids. By playing-back natural signals, we found that the two species as well as hybrid males responded to Alectoris signals but not to another species belonging to the Phasianidae family (Colinus virginianus). Results also indicate that red-legged and rock partridges responded stronger to conspecific calls than to heterospecific ones. However, they reacted similarly to conspecific and hybrid calls. F1 hybrids responded stronger to hybrids calls than to the two species ones. They did not distinguish the two parental species signals from each other. Although the two species showed the ability to discriminate the conspecific from the heterospecific signal, they clearly responded to the other species. This behaviour may play a role in the hybridization phenomenon. PMID- 11390088 TI - Human face recognition in sheep: lack of configurational coding and right hemisphere advantage. AB - Face recognition in sheep is qualitatively similar to that in humans in terms of its left visual field bias, and the effects of expertise and configural coding. The current study was designed to determine whether such effects are species specific by investigating the case of sheep recognising humans. It was found that the sheep could identify human faces and while they showed a small inversion induced decline in discriminatory performance, this was significantly less than seen with sheep faces. In other aspects, there were qualitative differences with human face recognition compared with conspecific recognition. In contrast with sheep faces there was no left visual field advantage in the recognition of human faces and the internal features were not used at all as visual cues. The data suggest that these sheep, whilst being extensively exposed to interactions with humans, were unable to identify them with all the same 'expert' methods as were used to discriminate other sheep. This suggests that different neural systems may, to some extent, be used for recognition of sheep as opposed to human faces. The relative contribution to differential neural processing of the faces of the different species and the role of expertise are discussed. PMID- 11390089 TI - Long-duration whirling of Pholcus phalangioides (Araneae, Pholcidae) is specifically elicited by Salticid spiders. AB - Long-duration whirling (gyrating of the body during several hours a day) was shown by the pholcid spider Pholcus phalangioides to salticid spiders and hardly ever to predatory spiders from eight other families in laboratory arenas. Long duration whirling has not been reported so far, in contrast to short-duration whirling lasting less than a few minutes. Long-duration whirling may have the anti-predatory function of disturbing continual visual fixation of prey in attacking salticids, in contrast to short-duration whirling that has been demonstrated to favour survival of pholcids in the presence of all sorts of predatory spiders. PMID- 11390090 TI - Nonlinear time perception. AB - Sensitivity to time was investigated to test the linear-timing hypothesis. A long duration was adjusted until accuracy was 75% correct for a short duration in a two-choice procedure. Short durations (2, 4, 6, 8, 10, 12, 14, 16 and 18 s) were selected from previous research that suggests that sensitivity to time is nonlinear in this range. Rats were tested with a single short interval (Experiment 1, n=13) or a random order (Experiment 2, n=7). A local maximum in sensitivity (d' from signal detection theory) was observed at approximately 8-12 s. Sensitivity to time was reliably correlated (r's=0.759-0.941) with previous data. Weber fractions exhibited a U-shape and were negatively correlated with sensitivity to time (r=-0.800). These results provide additional evidence that sensitivity to time is nonlinearly related to physical time. PMID- 11390091 TI - Visual categorization of natural and abstract items in forest monkeys and humans. AB - A preference for novelty paradigm was used to investigate whether mangabeys (Lophocebus albigena), an arboreal non-human primate species, were able to discriminate and to categorize different visual stimuli belonging to natural (food items) and abstract (non-food items) categories. In a comparative perspective human subjects were tested with the same procedure and the same stimuli. Two out of four mangabeys and three out of the four humans showed significant preference for novelty when comparing food versus non-food items. Hence they discriminated between these two sets of items. The two mangabeys and one non-adult human subject sorted the food items in one category, showing no preference for novelty when comparing known and unknown food-items and different views of the same food items. In contrast the two adult human subjects who showed preference for novelty in the between-category, did not show preference for novelty when comparing known and unknown food-items but did show such a preference when comparing different views of the same food items. Compared to human performances, the results suggest that mangabeys are able to form at least a perceptual category of natural, ecologically relevant stimuli. PMID- 11390103 TI - Molecular characterization of foot-and-mouth disease virus isolated from ruminants in Taiwan in 1999-2000. AB - In 1999, 10 sporadic outbreaks of cattle foot-and-mouth disease (FMD) occurred in Taiwan. By the time, infection was limited to the Chinese yellow cattle (a native species of beef cattle in Mainland China), which did not develop vesicular lesions under field conditions. Five viruses isolates obtained from individual farms were confirmed to be the serotype O FMD virus (O/Taiwan/1999). During January-February 2000, however, this virus has spread to dairy cattle and goat herds, causing severe mortality in goat kids and vesicular lesions in dairy cattle. Partial nucleotide sequence of the capsid coding gene 1D (VP1) was determined for the virus isolates obtained in this study. Phylogenetic analysis of the VP1 sequences indicated that the O/Taiwan/1999 viruses shared 95-97% similarities to the virus strains isolated from the Middle East and India. The species susceptibility of the O/Taiwan/1999 virus was experimentally studied in several species of susceptible animals, showing that the virus did cause generalized lesions in dairy cattle and pigs, however, it would not cause vesicular lesions on the Chinese yellow cattle and the adult goats. These studies suggested that the O/Taiwan/1999 virus was a novel FMD virus of Taiwan and it presented various levels of susceptibility in cattle species. PMID- 11390104 TI - Improved antigen and nucleic acid detection in a bovine virus diarrhoea eradication program. AB - A bovine viral diarrhoea/mucosal disease (BVD/MD) control and eradication program was introduced in Lower Austria in 1996, according to the Swedish model. An important risk factor for BVD transmission under local conditions is communal grazing where susceptible pregnant cattle from several herds may be mixed with unrecognised persistently infected (PI) animals. A reliable system for identification of PI animals is therefore essential for BVD eradication and steps were taken to improve a commercially available antigen-capture ELISA (Ag-ELISA) by modifying the method for leukocyte preparation and adjusting the negative cut off value. A single-tube reverse transcriptase-polymerase chain reaction (RT-PCR) employing panpestivirus 324/326 primers targeting the 5'-untranslated region of the virus genome was also simplified and used on pooled blood samples to facilitate larger sample throughputs. RT-PCR positive pools were analysed individually to identify infected animals. Seven hundred eighty-six samples were tested by Ag-ELISA according to the instruction manual and 5324 samples with the modified method. All 6110 samples were retested by RT-PCR. The percentage of RT PCR positive results with doubtful and negative Ag-ELISA samples significantly diminished using the modified method (from 4.71 to 0.82%). Selected BVD viruses were genetically typed by PCR product sequencing; special attention being paid to RT-PCR amplicons from samples which were negative or doubtful by ELISA. However, no correlation was found between the phylogenetic grouping of the viruses and the Ag-ELISA results. PMID- 11390105 TI - Deletion of gene 52 encoding glycoprotein M of equine herpesvirus type 1 strain RacH results in increased immunogenicity. AB - The immunogenicity of equine herpesvirus type 1 (EHV-1) strain RacH was compared to a RacH virus in which gene 52 encoding glycoprotein M (gM) was interrupted by insertion of LacZ (HDeltagM-Ins) and a RacH with 75% of gene 52 was deleted and replaced by LacZ (HDeltagM-HS). HDeltagM-Ins failed to produce full-length gM, but the carboxy-terminal portion was still expressed. No gM expression was detected in HDeltagM-HS-infected cells. Mice were immunised once with 1x10(3) to 1x10(5) plaque-forming units (PFU) of RacH or mutant viruses and challenged with virulent RacL11 virus 29 days later. A dose-dependence of protection was observed in RacH-immunised mice, and following immunisation with 1x10(4) or 1x10(3) PFU body weight losses and increased virus titres in lungs were observed after challenge infection. HDeltagM-HS-immunised mice were completely protected even after immunisation with 1x10(3) PFU. Mice immunised with 1x10(3) PFU of HDeltagM Ins but not the higher doses showed signs of disease after challenge infection. PMID- 11390106 TI - Deletions in the 7a ORF of feline coronavirus associated with an epidemic of feline infectious peritonitis. AB - A population of Persian cats experienced an epidemic of feline infectious peritonitis (FIP) over 2 years. Twelve cases of FIP occurred in litters born during this period. Cats contracting FIP were all genetically related through the sire. Feline coronavirus (FCoV) genomic RNA was detected consistently in this study in biologic samples from adult cats, kittens suffering from FIP, and their siblings. Analysis of viral 7a/7b open reading frame (ORFs) were analyzed and revealed two distinct virus variants circulating in the population, one with an intact 7a ORF and one with two major deletions in the 7a ORF. The 7b ORFs were intact and similar among all virus isolates, although point mutations resulting in amino acid changes were present. The sire was determined to be infected with both variants, and was persistently virus-infected. We speculate the deletion variant arose from the non-deletion variant during viral replication in this population, possibly in the sire. PMID- 11390107 TI - Analysis of the functional domains of Arcanobacterium pyogenes pyolysin using monoclonal antibodies. AB - Pyolysin (PLO), secreted by Arcanobacterium pyogenes, is a novel member of the thiol-activated cytolysin (TACY) family of bacterial toxins. Four monoclonal antibodies (mAbs) to PLO were prepared for the analysis of functional domains of this toxin. Two (mAbs S and H) of these markedly inhibited the hemolytic activity of PLO, but the inhibiting activity of the other two antibodies (mAbs C and G) was weaker. Subsequently, nine truncated PLOs were derived from recombinant Escherichia coli by various deletions from the N-terminus. Strong hemolytic activity was recognized in truncates of PLO following the deletion of 30 or 55 amino acids, but not in the truncate with deletion of 74 residues. Truncated PLOs were used in immunoblotting experiments to locate the epitopes for the mAbs. The epitope for mAbs C and G lies within the undecapeptide region (amino acids 487 505) of the C-terminus of PLO, which seems to be the binding site to erythrocytes. In contrast, the epitopes for mAbs S and H, which showed strong neutralizing activity, were found to lie in the N-terminal regions of the PLO ranging from 55 to 73 and 123 to 166 amino acids, respectively. From these results, it seems that the N-terminal region of PLO, in particular, the region of amino acids 55-74 is important for hemolytic activity. PMID- 11390108 TI - Phenotypic and genetic characterization of NAD-dependent Pasteurellaceae from the respiratory tract of pigs and their possible pathogenetic importance. AB - Nicotinamide adenine dinucleotide (NAD)-dependent Pasteurellaceae other than Actinobacillus pleuropneumoniae and Haemophilus parasuis are frequently isolated from the respiratory tract of pigs. The taxonomic classification and relevance for pathogenicity of these bacteria deserves further attention. In the present study, 107 of these NAD-dependent isolates from the porcine respiratory tract, primarily from lungs with pathological changes, were investigated. On the basis of phenotypic criteria, such as haemolysis, urease, catalase, and indole formation as well as other fermentative activities, 50 of the isolates were assigned to Actinobacillus minor, 36 isolates to Actinobacillus porcinus and 21 isolates to Actinobacillus indolicus. However, many isolates among the three species showed fermentative activities differing from those of the respective type strain of the species. Serotyping on the basis of heat-stable polysaccharide antigens and 16 rDNA sequencing also revealed substantial heterogeneity within each of the three species although they clustered together in three distinct groups in the phylogenetic analysis. These three groups of NAD-dependent bacteria are different from, or in a borderline position, to the existing species or genera within the family Pasteurellaceae. A considerable number of isolates of these three groups were isolated in pure cultures from pneumonic lungs. Consequently, it will be necessary to critically review the opinion, that these NAD-dependent Pasteurellaceae are only "agents colonizing the mucosa". Further, taxonomic examinations of the strains within these three groups are indispensable to testing isolates for their virulence in gnotobiotic pigs. PMID- 11390109 TI - Detection of Mycobacterium avium subsp. paratuberculosis in tissue samples by single, fluorescent and nested PCR based on the IS900 gene. AB - The aim of this study was to determine if fluorescent PCR could be used instead of nested PCR, for the detection of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) in clinical specimens, to improve the sensitivity without increasing the risk for cross-contamination. We investigated and compared the sensitivity of single PCR, fluorescent PCR and nested PCR for the detection of IS900, an insertion sequence specific for M. paratuberculosis. A previously described extraction method for clinical specimens, based on xylene, was evaluated regarding its suitability for routine diagnostic work. The sensitivity of each PCR system was assessed by analysing a serial dilution of M. paratuberculosis DNA. To improve the reliability of the PCR and to facilitate the interpretation of the PCR results, a positive internal control molecule ("mimic") was developed and used for single and fluorescent PCR. In nested PCR, an existing mimic was used. The efficiency of recovering DNA of M. paratuberculosis from clinical specimens by the extraction method and detection of the organism by PCR was studied by analysing spiked ileum mucosa specimens. The final evaluation was performed on seventeen ileum mucosa specimens, previously found positive for M. paratuberculosis by bacterial culture. Twelve of the samples were positive by fluorescent PCR and nested PCR, and 10 samples were positive by single PCR. The use of mimics showed inhibition in specimens harbouring few M. paratuberculosis organisms, illustrating the effect of inhibitory substances in combination with small amounts of M. paratuberculosis DNA. We conclude that the extraction method was not adequate to recover small amounts of M. paratuberculosis and that inhibitory substances were still present in the processed specimens, but that the method is useful for identifying positive samples. Fluorescent PCR was a suitable alternative to both single PCR and nested PCR for the detection of M. paratuberculosis. PMID- 11390110 TI - Recovery of Mycobacterium avium subsp. paratuberculosis from nematode larvae cultured from the faeces of sheep with Johne's disease. AB - A study was conducted to determine whether trichostrongylid nematode larvae become contaminated with Mycobacterium avium subsp. paratuberculosis when they develop in the faeces of sheep with Johne's disease. Nematode larvae were hatched from ova in the faecal samples of affected sheep. Larval sheaths were removed and these as well as exsheathed larvae were subjected to radiometric culture for M. paratuberculosis. The organism was recovered from washing water used to prepare the larvae, third stage larvae and larval sheaths, but not from exsheathed larvae. The recovery of M. paratuberculosis from larvae was associated with the severity of the histological lesions in affected sheep and with the results of culture of the organism from intestinal tissues and faeces. Nematode parasites of sheep might be able to act as mechanical vectors for M. paratuberculosis as the organism associates with infective third stage larvae when these develop in the faeces of sheep with Johne's disease. PMID- 11390111 TI - An insight into the epidemiology of dolphin morbillivirus worldwide. AB - Serum samples from 288 cetaceans representing 25 species and originating from 11 different countries were collected between 1995 and 1999 and examined for the presence of dolphin morbillivirus (DMV)-specific antibodies by an indirect ELISA (iELISA) (N = 267) or a plaque reduction assay (N = 21). A total of 35 odontocetes were seropositive: three harbour porpoises (Phocoena phocoena) and a common dolphin (Delphinus delphis) from the Northeastern (NE) Atlantic, a bottlenose dolphin (Tursiops truncatus) from Kent (England), three striped dolphins (Stenella coeruleoalba), two Risso's dolphins (Grampus griseus) and a bottlenose dolphin from the Mediterranean Sea, one common dolphin from the Southwest (SW) Indian Ocean, three Fraser's dolphins (Lagenodelphis hosei) from the SW Atlantic, 18 long-finned pilot whales (Globicephala melas) and a bottlenose dolphin from the SW Pacific as well as a captive bottlenose dolphin (Tursiops aduncus) originally from Taiwan. The presence of morbillivirus antibodies in 17 of these animals was further examined in other iELISAs and virus neutralization tests. Our results indicate that DMV infects cetaceans worldwide. This is the first report of DMV-seropositive animals from the SW Indian, SW Atlantic and West Pacific Oceans. Prevalence of DMV-seropositives was 85.7% in 21 pilot whales from the SW Pacific and both sexually mature and immature individuals were infected. This indicates that DMV is endemic in these animals. The same situation may occur among Fraser's dolphins from the SW Atlantic. The prevalence of DMV-seropositives was 5.26% and 5.36% in 19 common dolphins and 56 harbour porpoise from the NE Atlantic, respectively, and 18.75% in 16 striped dolphins from the Mediterranean. Prevalence varied significantly with sexual maturity in harbour porpoises and striped dolphins; all DMV-seropositives being mature animals. The prevalence of seropositive harbour porpoise and striped dolphins appeared to have decreased since previous studies. These data suggest that DMV is not endemic within these populations, that they are losing their humoral immunity against the virus and that they may be vulnerable to new epidemics. PMID- 11390112 TI - Survival of Mannheimia (Pasteurella) haemolytica in tracheobronchial washings of sheep and cattle. AB - The growth, morphology and long-term survival of a representative isolate of Mannheimia haemolytica serotypes A1 and A2 were monitored in ovine and bovine tracheobronchial washings. Both strains survived for at least 244 days in ovine tracheobronchial washings and 156 days in bovine tracheobronchial washings. The addition of fresh washings at these times prompted an increase for serotype A2 but no change in viability for serotype A1 in ovine tracheobronchial washings and an increase for both serotypes in bovine tracheobronchial washings. When growth and survival was compared using tracheobronchial washings from ruminant and non ruminant species there was a trend towards longer survival in ruminant fluids.Long-term survival was associated with temporary or permanent change from normal size colonies to 'micro-colonies' on sheep blood agar. Subculture allowed reversion to normal colony morphology. Analysis showed these micro-colonies to consist of chains of elongated bacteria. M. haemolytica serotype A2 was more robust in its ability to withstand nutrient deprivation for long periods of time. These survival mechanisms may have important implications for pathogenesis. PMID- 11390113 TI - Necrotoxigenic Escherichia coli type-2 invade and cause diarrhoea during experimental infection in colostrum-restricted newborn calves. AB - There exists experimental evidence that necrotoxigenic Escherichia coli (NTEC) strains producing the cytotoxic necrotising factor 1 cause intestinal and extra intestinal disease in piglets. On the other hand, no experimental model has been developed with NTEC strains producing the cytotoxic necrotising factor 2. In all, 14 colostrum-restricted calves were orally challenged with two strains isolated from the faeces of a diarrheic calf (B20a) or from the heart blood of a septicaemic calf (1404). All calves had diarrhoea which lasted until euthanasia in eight of them. In those calves, diarrhoea was correlated with the faecal excretion of the challenge strains. At necropsy, vascular congestion of the intestinal mucosa, hypertrophy of the mesenteric lymph nodes (MLN) and some congestion of the lungs were observed. Bacteriology confirmed the colonisation of the intestine by the challenge strains which were also recovered from the heart blood, the lungs and/or the liver. Histological sections confirmed enterocolitis, lymphadenitis and limited bronchopneumonia. In the intestinal tissue sections, bacteria testing positive in an in situ DNA hybridisation assay with a CNF2 probe were observed. Those results were confirmed by immunohistochemistry with a polyclonal anti-O78 and a monoclonal anti-F17b antisera. Three of the five control calves receiving either saline or a CNF(-), F17a strain (25KH09) had no clinical signs or lesions. The other two presented a profuse liquid diarrhoea but those calves were positive for the presence of K99(+) E. coli. In this model, both NTEC2 strains were thus, able to colonise the intestine, to cause long lasting diarrhoea and to invade the blood stream with localisation in various internal organs in colostrum-restricted conventional newborn calves. PMID- 11390114 TI - Passive immunization of pigs against experimental infection with Streptococcus suis serotype 2. AB - The safety and protective efficacy of a horse antiserum raised against inactivated whole cell preparations of Streptococcus suis serotype 2 was investigated in pigs by experimental challenge. The antiserum was evaluated in two similar experiments each comprising 12 4-week-old pigs treated with 6 ml of antiserum the day before challenge and four pigs used as challenge controls. Pigs were infected by subcutaneous injection with approximately 10(11) colony forming units of S. suis serotype 2. Clinical disease in the pigs that could be attributed to infection with S. suis was reduced from 88 to 35% (P = 0.015). The percentage of pigs with lesions that could be associated with S. suis was reduced from 88 to 22% (P = 0.002) and isolation of S. suis serotype 2 was reduced from five (63%) out of eight pigs in the combined challenge control groups to 3 (13%) out of 23 pigs in the combined treatment groups. These results indicate that passive immunization of pigs may be a way to reduce or control S. suis serotype 2 infections in pigs. PMID- 11390115 TI - Secreted antigens as virulence-associated markers in Staphylococcus aureus strains from rabbits. AB - Western blot analysis was performed from the culture supernatant of 59 rabbit Staphylococcus aureus strains, classified as high and low virulence strains according to their epidemiological behaviour in commercial rabbitries, bio-, phage- and RAPD-type. Fourteen extracellular antigen bands (A-N) were recognised using sera of rabbits immunised with washed, viable high virulence S. aureus bacteria. Eleven of these bands were found in high virulence as well as in low virulence strains. The band A, approximately 78 kDa, was not seen in any of the 27 high virulence strains, except for one strain which was also typical in other aspects, was detected in all, but one of the low virulence strains. The M and N bands with molecular masses of approximately 29 and 27 kDa, respectively, were recognised in all high virulence strains except for the atypical strain, but in none of the low virulence strains. This indicates that the latter two antigens may be virulence-associated markers for S. aureus strains from rabbits. PMID- 11390116 TI - The clinical, pathological and microbiological outcome of an Escherichia coli O2:K1 infection in avian pneumovirus infected turkeys. AB - The purpose of this study was to evaluate the effect of an Escherichia coli infection in avian pneumovirus (APV)-infected turkeys. One group of 2-week-old specific pathogen-free (SPF) and two groups of 3-week-old conventional (CON) turkeys were inoculated oculonasally with virulent APV subtype A alone, with E. coli O2:K1 alone or with both agents at varying intervals (1, 3, 5 or 7 days) between the two inoculations. The birds were followed clinically and examined for macroscopic lesions at necropsy. Titres of APV were determined in the turbinates, trachea, lungs and air sacs. The number of E. coli O2:K1were assessed in the turbinates, trachea, lungs, air sacs, liver and heart. In both SPF and CON turkeys, dual infection resulted in an increased morbidity and a higher incidence of gross lesions compared to the groups given single infections, especially with a time interval between APV and E. coli inoculations of 3 and 5 days. APV was isolated from the respiratory tract of all APV-infected groups between 3 and 7 days post inoculation. E. coli O2:K1 was isolated only from turkeys that received a dual infection. It was recovered from the turbinates, trachea, lungs, heart and liver. These results show that APV may act as a primary agent predisposing to E. coli colonization and invasion. PMID- 11390117 TI - Genetic typing of bovine viral diarrhea virus isolates from Argentina. AB - Genetic typing of 29 Bovine Viral Diarrhea Virus (BVDV) isolates from Argentina was carried out by sequencing 245 nucleotides of the RT-PCR products of the 5' UTR region. Sequence analysis shows that these Argentinean BVDV include types 1 and 2. The majority (26/29) of the isolates are type 1, which comprises subtypes 1a and 1b, together with an additional subgroup within subtype 1a. This subgroup is close to the South African subgroup Ic of 1a viruses, and to the deer pestivirus strain "Deer". The three type 2 BVDV were isolated from fetal tissues or serum during the 7-8 years before a clinical outbreak in Argentina had been reported. Only inactivated vaccines are used in bovines of the country, thus the analysed viruses are authentic field strains. The long term circulation of type 2 BVDV (situation similar to that of North America before the epidemic of 1993), and the existence of viral populations which differ from the reference strains commonly used in vaccine elaboration should be considered by manufacturers of diagnostic reagents and vaccines. PMID- 11390118 TI - Hormone replacement therapy for the postmenopausal woman. PMID- 11390119 TI - Cellular estrogen activity: implications for pulsed estrogen therapy. AB - Estrogens exert their principal biological effects through the actions of two different intracellular estrogen receptor (ER) proteins, ER alpha and ER beta. Following the binding of steroid, the protein undergoes a conformational change that results in a transcriptionally active form. The receptor protein is locked into an active state by estradiol, which results in the transition of the receptor through a signal transduction cascade of events, ultimately resulting in the activation of specific genes, thereby inducing the biological events specific for that type of target cell. There is a large variation in the relative expression levels of the two ER isoforms in different target tissues and in different stages of development. In addition, variant forms of the two ER isoforms, the result of splice variation, have been described. ER alpha and ER beta have been shown to differ in specific aspects within the various stages of the signal transduction pathway. Thus, there is a broad spectrum of estrogen response mechanisms as a result of an infinite number of possible combinations of all these factors. In addition, there are gene regulatory mechanisms that are the result of ER--protein interactions instead of ER--DNA interactions. Steroid binding is the key initiating action of the whole pathway, which, in terms of cell biology, is a relatively slow process. The response induced through the action of ER induction can be shown to be dependent on the total dose exposure rather than estradiol concentrations at subsaturating levels. PMID- 11390120 TI - Dose-ranging studies of a novel intranasal estrogen replacement therapy. AB - OBJECTIVE: To study the efficacy and tolerability of Aerodiol, a novel intranasal estradiol spray, and to determine an appropriate dose range. METHODS: An exploratory, parallel-group, dose-finding study was followed by a large-scale, double-blind, placebo-controlled study. In the exploratory study, 134 postmenopausal women were allocated to receive a daily dose of 100--900 microg of Aerodiol for 12 weeks. Efficacy was determined by an overall assessment of estrogenization. In the placebo-controlled study, 420 postmenopausal women were randomized to receive 100, 200, 300, or 400 microg of Aerodiol, or oral estradiol 1 or 2 mg, or placebo, daily for 12 weeks. Efficacy was assessed by the Kupperman Index and the number of hot flushes per day after 12 weeks. RESULTS: In the exploratory study, the level of estrogenization was sufficient for 23% of women in the 100 microg/day group, excessive for 36% of the 900 microg/day group, and good for more than 80% of women receiving 200--600 microg/day. In the placebo controlled study, the Kupperman Index at week 12 was significantly lower than placebo (P<0.01) for all Aerodiol groups except the 100 microg/day group. The efficacy of Aerodiol 300 microg/day was similar to oral estradiol 2 mg/day. The variability in exposure to estradiol was lower with Aerodiol than with oral estradiol. Aerodiol therapy was well tolerated in both studies. Premature withdrawals were approximately equally distributed among treatment groups in the placebo-controlled study. CONCLUSIONS: Aerodiol was effective in reducing climateric symptoms at doses between 100 and 600 microg/day. Treatment was well tolerated and well accepted. A dose of 300 microg/day is recommended for initiating hormone replacement therapy. PMID- 11390121 TI - Therapeutic value and long-term safety of pulsed estrogen therapy. AB - OBJECTIVES: To demonstrate equivalent efficacy for menopausal symptoms between Aerodiol nasal spray and reference oral estradiol therapy, and to investigate the endometrial safety and tolerability of Aerodiol in the long term. METHODS: The efficacy of Aerodiol 300 microg, once daily, was compared with oral estradiol 2 mg/day in a randomized, double-blind trial. A statistical test of noninferiority was performed on the mean absolute Kupperman index (KI) obtained after 14 and 23 weeks of the two treatments. Long-term safety was assessed in a 1-year open-label study. The initial Aerodiol dose was 300 microg/day, and was adjusted if required. Endometrial biopsies were obtained at inclusion and at the end of the trial and examined independently by two pathologists. RESULTS: In the equivalence trial, the KI improved similarly in the Aerodiol group (n=317) and the oral estradiol group (n=342). Aerodiol was shown statistically to be at least as effective as oral therapy (P<0.001), but the incidences of mastalgia and withdrawal bleeding were significantly lower in the Aerodiol group (P<0.01 and P<0.001, respectively). In the long-term safety trial (n=408), the rate of Aerodiol treatment continuation at 12 months was 85%, and there was no incidence of endometrial hyperplasia or cancer. Aerodiol dose adaptation was performed by 29% of women. CONCLUSIONS: Aerodiol was shown to have equivalent efficacy to reference oral estradiol therapy, but with better gynaecological acceptability. The endometrial safety of Aerodiol was confirmed in the long term, and the ability to adjust the dosage easily was of benefit to a substantial proportion of women. PMID- 11390122 TI - Aerodiol versus the transdermal route: perspectives for patient preference. AB - OBJECTIVE: to compare the efficacy, tolerability and user preference of Aerodiol intranasal and transdermal patch administration of 17 beta-estradiol for climacteric symptoms. METHODS: an open-label, multicenter, crossover trial in which recently postmenopausal women were randomized to receive either Aerodiol 300 microg daily (n=176), or transdermal 17 beta-estradiol (reservoir patches delivering 50 microg/day), 2 patches per week (n=185), for 12 weeks, followed by 4 weeks of the alternative treatment. Efficacy was assessed primarily by the Kupperman index at the end of each treatment period. User satisfaction was assessed by questionnaire at weeks 12 and 16, and at week 16 the women chose which treatment they preferred to use for a further 40-week period. RESULTS: Aerodiol and transdermal patch therapy produced marked and similar reductions in the Kupperman index and the incidence of hot flushes at weeks 12 and 16. The reduction in the Kupperman index at week 12 was statistically equivalent for the two treatments. The tolerability of both treatments was good, with similar numbers of emergent adverse events reported in both groups. The incidence of moderate or severe mastalgia, however, was significantly lower with Aerodiol (P=0.02). Significantly more women chose to continue treatment with Aerodiol than with the transdermal patch (66 vs. 34%, P<0.001). When all women had experienced both treatments, reported levels of satisfaction were significantly higher with Aerodiol than with transdermal therapy (P<0.001 for all six categories assessed). CONCLUSIONS: Aerodiol and transdermal patch treatments were of similar efficacy and tolerability. Levels of user preference and satisfaction, however, were higher with Aerodiol, which should contribute towards good long-term compliance with this therapy. PMID- 11390123 TI - Perspectives in hormone replacement therapy. AB - Estrogens have been convincingly shown to be highly effective in preventing and reversing menopause-related conditions, such as hot flushes, urogenital complaints, and postmenopausal bone loss. Observational studies report that long term, estrogen-containing, postmenopausal hormone replacement therapy (HRT) leads to a substantial reduction in hip fractures, myocardial infarction, and possibly colonic cancer, with important consequences for health and quality of life. Estrogen replacement may postpone the onset of Alzheimer's disease and extend life. While many of these effects are biologically plausible, with a variety of cellular mechanisms being involved, only ongoing and future large-scale randomized clinical trials can and should define the effects of HRT more precisely. Long-term compliance is a key issue for long-term benefits, and offering women a choice of administration routes and regimens can only be beneficial in this respect. Pills, patches, gels, and implants are all widely prescribed. Intravaginal or intranasal forms of administration, which are very easy to use and adaptable on an individual level, are among the new options which could improve long-term continuation of HRT use. Fear of breast cancer and recurrence of vaginal bleeding are real concerns for many women considering HRT. This has led to research into lower-dose, estrogen-containing regimens, into continuous combined regimens, and into the potential of estrogen receptor alpha or beta binding molecules that may help to prevent such problems from arising. The prospects for safe and effective postmenopausal HRT with either estrogens or estrogen-like drugs are very promising when these drugs are used in a patient tailored, risk profile-based manner. PMID- 11390124 TI - HRT in the third millennium. PMID- 11390125 TI - Journal of Ethnopharmacology: An interdisciplinary journal devoted to indigenous drugs. PMID- 11390126 TI - Treatment with extracts of Momordica charantia and Eugenia jambolana prevents hyperglycemia and hyperinsulinemia in fructose fed rats. AB - Insulin resistance has been implicated as a major contributor to the development of hyperglycemia in NIIDM patients. Herbal extracts of Momordica charantia (MC) and Eugenia jambolana (EJ) have been shown to reduce hyperglycemia in diabetic animal models and human patients. However, no work has been done so far to assess their effect on insulin resistance. This study was undertaken to study the effects of different doses (100,200 and 400 mg per day) of alcoholic and aqueous extracts of MC and EJ on the metabolic parameters (body weight and serum glucose, insulin and triglycerides levels) of fructose fed rats. Fructose feeding for 15 days increased serum glucose and insulin levels markedly and triglycerides levels marginally vs. control (75.46+/-2.41 vs. 55.59+/-2.89 mg/dl, 6.26+/-1.27 vs. 15.04+/-2.43 mg/dl and 50.93+/-3.30 vs.41.1+/-3.33 mg/dl, respectively). Treatment with 400 mg per day of aqueous extracts of MC and EJ for 15 days substantially prevented hyperglycemia and hyperinsulinemia induced by a diet high in fructose (63.52+/-2.9 and 66.46+/-2.2 vs. 75.46+/-2.4, respectively). PMID- 11390127 TI - Anticonvulsant properties of the methanolic extract of Cyperus articulatus (Cyperaceae). AB - The methanolic extract of rhizomes of Cyperus articulatus, a plant used in traditional medicine in Africa and Latin America for many diseases, possesses anticonvulsant activity in mice. This extract protected mice against maximal electroshock (MES)- and pentylenetetrazol (PTZ)-induced seizures. It also delayed the onset of seizures induced by isonicotinic acid hydrazide and strongly antagonized N-methyl-D-aspartate-induced turning behavior. The ED(50) for protection against seizures was 306 (154-541) mg/kg intraperitoneally (i.p.) for the PTZ test and 1005 (797-1200) mg/kg i.p. for the MES test. The ED(50) of methanolic extract against N-methyl-D-aspartate-induced turning behavior was 875 (623-1123) mg/kg i.p. C. articulatus L. methanolic extract protected 54% of mice from seizures induced by strychnine at the dose of 1000 mg/kg i.p. but had no or a moderate effect only against picrotoxin- or bicuculline-induced seizures. With these effects, the rhizome of C. articulatus L. possesses anticonvulsant properties in animals that might explain its use as a traditional medicine for epilepsy in Africa. PMID- 11390128 TI - Toxicity studies in mice of Commiphora molmol oleo-gum-resin. AB - Acute (24 h) and chronic (90 days) oral toxicity studies on Commiphora molmol (oleo-gum-resin) were carried out in mice. Dosages in acute study were 0.5, 1.0 and 3 g/kg, while in chronic study dosage was 100 mg/kg per day. All external morphological, biochemical and haematological changes, in addition to body and vital organ weights were recorded. There was no significant difference in mortality in acute or chronic treatment as compared to controls. At the end of the treatment, weight gain in the treated as well as control group was significant. There was a significant increase in weight of testes, caudae epididymides and seminal vesicles in C. molmol treated group. Biochemical studies revealed no differences in C. molmol treated animals, however, haematological studies revealed a significant increase in RBC and haemoglobin levels as compared to the control group. C. molmol failed to show any spermatotoxic effects. PMID- 11390129 TI - Anti-nociceptive and anti-inflammatory properties of Caralluma arabica. AB - In the present study, a 10% ethanolic extract of Caralluma arabica at the doses of 200 and 400 mg/kg was evaluated for the antinociceptive activity using the hot plate and acetic acid induced abdominal constrictions in mice and tail flick method in rats. Oral and topical application of C. arabica extract was assessed in the present study for anti-inflammatory properties using carrageenan-induced rat paw oedema. Anti-inflammatory activity was also evaluated using cotton pellet granuloma method. C. arabica extract showed significant Anti-nociceptive properties in all the models studied. C. arabica extract significantly reduced the increase in hind paw volume induced by carrageenan injection when used orally or topically. Results of the present study confirm the use of C. arabica traditionally for the treatment of painful and inflammatory conditions and indicate transdermal absorption of the plant extract. PMID- 11390130 TI - Effects of the flavonoids extracted from Spergularia purpurea Pers. on arterial blood pressure and renal function in normal and hypertensive rats. AB - The antihypertensive and diuretic effects of the flavonoids extracted from Spergularia purpurea Pers. (SP) were studied both in normotensive (NTR) and spontaneously hypertensive conscious rats (SHR). Daily oral administration of the flavonoid mixture (5 mg/kg for 1 week) exhibited a significant decrease in blood pressure with variation coefficient (Delta) of 20 in SHR rats and 11 in NTR rats. The systolic and diastolic blood pressure decreased significantly and respectively with 17 and 24% in SHR, and with 11 and 16% in NTR. The flavonoid mixture enhanced significantly the water excretion in hypertensive (P<0.001) and normal rats (P<0.001). Furthermore, oral administration of flavonoids mixture at a dose of 5 mg/kg produced a significant increase of urinary excretion of sodium (P<0.01), potassium (P<0.05) and chlorides (P<0.05) in SHR. Similarly, the flavonoid administration induced a significant increase of urinary electrolytes elimination in NTR (P<0.01 versus controls). No significant changes were noted on heart rate after flavonoids treatment in SHR as well as in NTR. While, glomerular filtration rate showed a significant increase after administration of flavonoids in all groups (P<0.05). These results suggest that oral administration of flavonoids obtained from Spergularia purpurea exhibited antihypertensive and diuretic actions. PMID- 11390131 TI - Antioxidant activity of Argentine propolis extracts. AB - Propolis is used in Argentine folk medicine. We have examined its possible protective action against oxidative modification of lipid in unfractionated serum. The kinetics of copper-induced oxidation was continuously monitored by measuring the formation of conjugated dienes, as the increase in the absorbance at 234 nm. According to the kinetics of oxidation, the propolis were classified in three different groups. Group I (CE, CO, BO, MO, BE) inhibited lipid oxidation during the initiation and propagation phases even at low concentrations. Group II (SP, CA, AM) increased the lag-phase for conjugated diene formation. All propolis in groups I and II diminished the maximal rate of diene production and the maximal amount of dienes produced. Group III (PA, RA, FE, VR, TV) had no effect on the lipid oxidation. The extent of lipoprotein oxidation was measured by the thiobarbituric acid reactive substance assay. Generation of malondialdehyde-like substances was inhibited and delayed by the presence of propolis extracts from group I and II. Our results justify the use of propolis (groups I and II) as a source of natural antioxidants. PMID- 11390132 TI - Neuropharmacological actions of Portulaca oleraceae L v. sativa (Hawk). AB - Portulaca oleracea L v. sativa (family: Portulacaceae) is a warm-climate annual, cultivated in the Arabian peninsula and used traditionally for alleviating pain and swelling. It was observed that a 10% ethanolic extract of this plant produced restriction of movement in animals during the routine screening studies. Therefore the effects of the extract on the locomotor activity, threshold to noxious stimulus, anti-convulsant activity and relaxant effects on the skeletal muscle were studied. The extract, on intraperitoneal administration, showed a significant reduction in the locomotor activity in mice, anti-nociceptive activity in rats using Tail Flick Method, an increase in the onset time of pentylenetetrazole-induced convulsions in mice and muscle relaxant activity in in vitro (rat hemidiaphragm) and in vivo (grip strength) experiments. The anti nociceptive activity of the extract in rats was attenuated by naloxone pre treatment indicating the involvement of opioid receptors in its anti-nociceptive effects. It is indicated from the results of the present study that P. oleracea v. sativa possesses varied effects on both the central and peripheral nervous system and the plant should be exhaustively studied for other neuropharmacological effects. PMID- 11390133 TI - Perspectives in ethnopharmacology: forging a closer link between bioscience and traditional empirical knowledge. AB - To what extent do ethnopharmacologists from diverse disciplines share a vision of what ethnopharmacology is and what it might become? This question was explored several years ago through content analysis of the Journal of Ethnopharmacology (JEP), the official journal of the International Society for Ethnopharmacology (ISE). The analysis revealed that although the published articles represent the breadth of natural and social sciences, most studies are themselves not synthetic or interdisciplinary. For the present study, analysis was extended through the most recently published issues of the JEP and compared, for the same time period, to the subject matter of another natural products journal, Pharmaceutical Biology. Whereas research published in the JEP better represents the interdisciplinary objectives of that journal, the difference is not striking. By way of illustration, several studies are reviewed that represent the unique, synthetic perspective that is highlighted in the mission statements of both the JEP and the ISE. The conclusion underscores the lack of clarity in research objectives and suggests that ethnopharmacologists of all backgrounds can enhance their work by projecting pharmacologic data against a backdrop of medical ethnography and by enriching cultural interpretations of medical actions by exploring the physiologic potential of plants. PMID- 11390134 TI - Antibacterial and antifungal activity of the essential oils of Thymus revolutus Celak from Turkey. AB - The chemical composition of the volatile constituent from flowering parts of Thymus revolutus C., an endemic plant of Turkey, were analysed by GC/MS. Twenty two components were identified, and carvacrol was found as a predominant compound in the oil. Furthermore, the essential oil was tested against 11 bacteria and four fungi at different concentrations. Results showed that the oil exhibited a significant antibacterial and antifungal activity. PMID- 11390135 TI - Hepatoprotective activity of Luffa echinata fruits. AB - The different extracts of the fruits of Luffa echinata Roxb. (Cucurbitaceae) were tested for their hepatoprotective activity against CCl(4) induced hepatotoxicity in albino rats. The degree of protection was measured by using biochemical parameters like serum glutamic oxalacetic transaminase (SGOT), serum glutamic pyruvate transaminase (SGPT), alkaline phosphatase (ALKP), total protein (TP) and total albumin (TA). The petroleum ether, acetone and methanolic extracts showed a significant hepatoprotective activity comparable with those of Silymarin. PMID- 11390136 TI - Antiulcer action of the hydroalcoholic extract and fractions of Davilla rugosa Poiret in the rat. AB - Davilla rugosa Poiret ("Cipo- Caboclo") is commonly used in Brazilian folk medicine. The hydroalcoholic extract of the stems (HE) was fractionated with chloroform (CF), chloroform/ethyl acetate (CAF), ethyl acetate (AF), ethyl acetate/ethanol (AEF), ethanol (EF), and ethanol/water (EWF). The hydroalcoholic extract (HE) and fractions of the stems of D. rugosa Poiret were investigated for possible anti gastric ulcer properties. These extracts were shown to protect rats from developing gastric ulcers induced by two acute models: HCl/ethanol (400 mg/kg i.p.) and immersion-restraint stress (15 and 30 mg/kg of the HE and 15 mg/kg either of the ethanol or the ethanol/water fractions, p.o.). The daily oral dose of 800 mg/kg of HE for 30 consecutive days was tested for possible toxic effects. There were no modifications in body weight, water or food intake or in the external aspect of kidneys, spleen, lungs and liver. The only difference observed was a decrease of liver weight. These results suggest that the D. rugosa Poiret HE has an antiulcer activity in rats and the active components are in the two more polar fractions. PMID- 11390137 TI - Antihistaminic and mast cell stabilizing activity of Striga orobanchioides. AB - The ethanolic and aqueous extracts of the whole plant of S. orobanchioides were evaluated for antihistaminic and mast cell stabilizing activities. Both extracts inhibited histamine-induced contractions of the guinea-pig ileum at the concentration range of 2.5-25 microg/ml in a dose-related manner. At 25 microg/ml, both extracts inhibited the response of histamine (0.5 microg/ml) almost completely. The effect of these two extracts on the degranulation rate of sensitized peritoneal cells of albino rats when challenged with antigen (horse serum) was studied. Triple vaccine was used as adjuvant. Ketotifen and prednisolone were used for comparison. The ethanolic extract at 100 and 200 mg/kg body weight was found to significantly inhibit degranulation of mast cells to an extent of 52.14+/-3.24 and 67.96+/-3.70%, respectively. At the same doses, the aqueous extract showed 42.09+/-2.91 and 60.67+/-3.50% reduction in degranulation of mast cells, respectively. Hence, both extracts markedly protected the rats against antigen-induced challenge of mast cells. PMID- 11390138 TI - The forensic DNA implications of genetic differentiation between endogamous communities. AB - In many indigenous minority populations, and among migrants from Asian and African populations now resident in western Europe, North America and Australia, there is a strong tradition of endogamy and a preference for consanguineous unions. These marriage practices can result in F(ST) values greatly in excess of the maximum value (0.01) currently recommended for forensic DNA purposes under guidelines established by the National Research Council (NRC) of the USA. To examine the possible extent of deviation from this accepted norm, three co resident Pakistani communities were studied using 10 autosomal dinucleotide markers and six tetranucleotide markers on the Y-chromosome. The mean population subdivision coefficient (FST) value was 0.13 for the autosomal loci, and Y chromosome loci exhibited even stronger differentiation with unique alleles identified in all three communities. The data indicate that even when sub populations are virtually indistinguishable in terms of anthropology, geography, ethnicity or culture, they may still exhibit major genetic differentiation. Where significant population stratification is known to exist, more detailed genetic databases should be developed for forensic DNA purposes, based on reference data from each of the appropriate sub-populations and not on random or combined samples. PMID- 11390139 TI - Microsatellite stability in human post-mortem tissues. AB - Human identification and forensic criminal casework may involve DNA profiling of decomposed material. Somatic microsatellite (STR) instability may lead to false exclusions and theoretically to false inclusions, both in criminal cases and in human identification. Hence, the somatic and postmortal stability of the actual sequences is crucial to the reliability of such analyses. Somatic STR stability in human tissues has been documented in small series only and the effect of postmortal tissue decomposition on microsatellite stability remains to be elucidated. On this basis, we have systematically searched for somatic STR mutations in 26 deceased humans without signs of decomposition at autopsy and 25 autopsy cases with obvious signs of postmortal decomposition. A blood sample and six tissue samples were collected from each case. Seven STRs were chosen for study, the tetranucleotides HUMVWA31/A, HUMTH01, HUMF13A1, and HUMFES/FPS, and the hyperpolymorphic markers HUMAPOAI1, D11S554 and HUMACTBP2. Denaturing gel electrophoresis was performed on an ABD Prism 377 gene sequencer with Genescan 672 software (Applied Biosystems, Inc.). The bone DNA profile of each case was chosen as the standard DNA profile. All cases gave profiles from additional tissues. By intraindividual comparison of DNA profiles in the cases without signs of degradation we find that the short repetitive sequences under study are stable, that is without evidence of somatic mutations. The cases with varying degree of decomposition display postmortal microsatellite stability, we detect no somatic mutations or other possible postmortal changes that could lead to between organ non-matches. In conclusion, PCR-based STR analyses are suitable in human identification and forensic casework dealing with different tissues, even when the substrate is heavily decomposed. PMID- 11390140 TI - Mortality in maximum security psychiatric hospital patients. AB - Numerous studies have mentioned to the high percentage of violent deaths in prison psychiatric hospitals, with suicide being the principal cause. The aim of this study was to analyze the circumstances related with the deaths recorded in a high security institution. Postmortem reports on all the deaths at the Alicante Psychiatric Prison between 1984 and 1997 were studied (36 cases of unnatural death and 28 of natural death). Of the violent deaths recorded 34 (94.4%) were suicides. Demographic, clinical and interpersonal variables factors were registered. In the cases of suicides, the method used, the place of death, season, month and time of suicide were analyzed. In our study, 64.7% of suicides were schizophrenic and 32.4% had inflicted self-harm previously. We found a statistically significant association between the cause of death (natural, suicide or homicide) and age, 47.1% of suicides being between the ages of 18 and 30 and 29.4% between the ages of 30 and 45. Natural causes predominated in older subjects. The prison population studied showed grave negative traits, mental illness and criminal behavior having forced them to the very edge of society. Our results were compared with the death and suicide rates of the general Spanish population. PMID- 11390141 TI - Prevalence of nicotine consumption in drug deaths. AB - One hundred consecutive drug death victims autopsied at the Institute of Forensic Medicine, University of Freiburg, between 1995 and 1997 were studied retrospectively as to whether the drug users had also consumed nicotine. The study included histological examination of the lung tissue for smoker cells and radioimmunological as well as GC-MS assays of the urine for cotinine, the primary metabolite of nicotine. It was found that 98 out of 100 drug victims had consumed nicotine in addition to illicit drugs or replacements. Yellowish-brown discolorations on the middle and index fingers were discernible in 44 drug victims, whereas fresh or scarred burns due to glowing cigarettes were found in six deceased drug consumers. Diseases of the bronchial system typical of heavy smokers were seen in 35 cases. Siderophages could be demonstrated in 17 of the 100 drug deaths. PMID- 11390142 TI - Examination of line crossings by atomic force microscopy. AB - Until now, the most widely used methods for the forensic examination of line crossings in documents were optical and electron microscopy. The combination of both techniques allows one in most cases to establish the sequence of lines. The recent development of scanning probe microscopy [1] gives an opportunity to complement or even replace the classical instruments used in this field. Scanning probe microscopes have been designed to study surfaces at high magnification. The aim of this study was to verify if their most popular member, the atomic force microscope (AFM) [2], can be applied to line crossing problems. The results show for the first time that AFM images present the same qualitative information obtained by scanning electron microscope (SEM) images and, consequently, allow the determination of the line crossing sequence under ambient conditions without vacuum and conductive coating of specimens. PMID- 11390143 TI - The short tandem repeat locus VWF2 in Intron 40 of the von Willebrand factor gene consists of two polymorphic sub-loci. AB - This study describes the complex nucleotide sequence structure of the TCTA short tandem repeat (STR) locus, VWF2. Eight alleles of VWF2 were observed in a population of 116 unrelated Caucasian individuals. The alleles ranged in size from 150 to 178 base pairs (bp). Sequence analysis of the isolated alleles revealed two polymorphic regions that were named sub-loci VWF2-a and VWF2-b. VWF2 a is located at the 5' end of the conventional locus, whilst VWF2-b is located at the 3' end. The two sub-loci are joined by a 30-nucleotide non-polymorphic sequence which contains two additional TCTA motif repeats. A semi-nested polymerase chain reaction (PCR) was designed to amplify the VWF2-b region in conjunction with the standard VWF2 amplification. This new amplification method enabled a higher level of allele discrimination than could be achieved by assigning alleles according to size. A cohort of 99 unrelated individuals was tested with this method. VWF2-a expressed five different alleles ranging from zero motif repeats to four motif repeats, while VWF2-b alleles ranged from 8 to 14 motif repeats. Allelic configuration based on the VWF2-a and VWF2-b sub alleles revealed 23 unique configurations out of a possible 31 for the original eight VWF2 alleles. In conclusion, the VWF2 is a highly polymorphic STR locus with potential application for forensic and parentage testing. PMID- 11390144 TI - Classification of prepaid cards based on multivariate treatment of data obtained by X-ray fluorescence analysis. AB - Two hundred prepaid cards, which had been used in Nagoya-city's subway in Japan, and another 32 prepaid cards (11 were real turnpike cards, 20 were counterfeit cards and 1 was a white card) were evaluated by X-ray fluorescence (XRF) without any pre-treatment. A preliminary investigation was performed on 200 prepaid cards in order to find an identification method for the turnpike cards. By plotting the relative intensity of titanium versus that of iron obtained by XRF, the cards were clearly classified into seven groups. On the other hand, the cards could be divided into four groups by a multivariate analysis using the relative intensities of five elements such as chlorine, calcium or tin, titanium, iron and barium. Using these results to classify the Japanese turnpike cards, they were divided into three groups or two groups. One of three groups or two groups was the counterfeit card group. PMID- 11390145 TI - Histopathological findings of cardiac conduction system of 150 Finns. AB - One hundred and fifty heart specimens were collected from the cases submitted for autopsy in the Department of Forensic Medicine in the University of Turku in March-May 1995 and May-July 1996, respectively. The cardiac conduction system (CCS) of these hearts were examined in order to find out the histopathological changes in the CCS of Finnish persons and their forensic pathological significance. There were 94 males and 56 females. Almost all age groups were included. The results revealed that in most of the persons the fibrous tissue and fatty tissue in the CCS increased with aging. In about half of the persons, there was deposition of calcium in the central fibrous body, pars membranacea, and the top of the musculature in the interventricular septum. In seven cases, the atrioventricular node (AVN), His bundle (HB) or bundle branches (RBB, LBB) were compressed by the calcium deposition. Hemorrhage, inflammation, amyloidosis, tumor, fatty infiltration and developmental malformations were observed in 31 cases. Twenty-eight cases died of myocardial infarct, among them, no involvement of the CCS could be observed. The authors concluded that routine examination of the CCS is helpful for revealing diseases of the CCS and improving the quality of forensic pathological diagnosis. PMID- 11390146 TI - SPME-GC analysis of THC in saliva samples collected with "EPITOPE" device. AB - In this study we examined the presence of cannabinoids in saliva samples obtained from 24 drug-abusers. The saliva specimens were collected by "EPITOPE" system and the subsequent elution of samples was achieved by centrifugation. The resulting ultrafiltrates have been directly sampled with solid phase micro-extraction (SPME) and then analyzed by GC/MS. Saliva sampling is less invasive than collection of blood. PMID- 11390147 TI - A light stabilizer Tinuvin 770-induced toxic injury of adult rat cardiac myocytes. AB - Tinuvin 770/bis(2,2,6,6-tetramethyl-4-piperidinyl)sebacate is a worldwide used light stabilizer for plastic materials like polyolefins. Tinuvin 770 is a biologically active component of polypropylene tubes. Glossmann and his study group managed to extract this compound by aqueous or organic solvents from laboratory plastic tubes, and propose that Tinuvin 770 is a potent blocker of L type Ca(2+)-channel through the phenylalkylamine and benzothiazepine-selective drug binding domains of the alpha(1) subunit of the receptor [Proc. Natl. Acad. Sci. U.S.A. 90 (1993) 9523]. We examined the direct morphological effect of Tinuvin 770 in give 25nmol, 0, 30, 60, 120 minute exposure time in isolated cardiomyocytes from adult rats. Incubation of myocytes with Tinuvin resulted in a progressive decline of rod-shaped and viable cells. It was accompanied by an increase in number of hypercontracted myocytes with microbleb formation compared to control and depletion of ATP level. In summary, our results demonstrate that plasma membrane damage and hypercontraction are manifestations of Tinuvin-induced injury of isolated cardiomyocytes. PMID- 11390148 TI - STR data for the GenePrint PowerPlex 1.2 system loci from three United Arab Emirates populations. AB - Allele frequencies for the eight STRs included in the GenePrint PowerPlex 1.2 kit were obtained from three population groups that are resident in the United Arab Emirates (UAE); 228 unrelated UAE individuals, 194 unrelated Indian individuals and 197 unrelated Pakistani individuals were analysed. PMID- 11390149 TI - Vaginorectal impalement injury in a 2-year-old child--caused by sexual abuse or an accident? AB - Anogenital impalement injuries are rarely encountered in clinical or forensic practice. If seen in children and if incurred under suspect circumstances, sexual abuse must be considered in the differential diagnosis. Here we describe the case of a 2-year-old girl admitted to hospital with a vaginorectal impalement injury. According to the girl's parents, she had slipped in the bathroom and fallen onto the handle of an upright toilet bowl brush. Since a second slight anal injury was present, the parents' account appeared inconsistent. Therefore, physicians from the Institute of Legal Medicine were consulted to investigate the possibility of underlying sexual abuse. Because they could not rule out that the injuries could have been caused by sexual abuse, they recommended having the police immediately examine the site of the purported accident for evidence. The police and forensic investigations, however, verified the parents' account of an accidental injury. Thus, in this case, the initiation of a police inquiry, which is not compulsory in Germany even when sexual abuse is strongly suspected, led to the exoneration of the father. PMID- 11390150 TI - Patient education in Europe: united differences. AB - This issue of Patient Education and Counseling presents the state of the art of patient education in several European countries. It is based on papers presented at a meeting in Paris on the evolution and development of patient education in western, central and eastern Europe (May 1999). Also patient education in the US is presented in this issue. Patient education is defined as all the educational activities directed to patients, including aspects of therapeutic education, health education and clinical health promotion. Five important factors are identified in the development of patient education: (1) research and evidence based standards; (2) the organization of care; (3) training and methodological support; (4) professional values; and (5) acknowledgment, funding and place of patient education in health policy. Several of the discussed common orientations and priorities in the patient education in the reviewed countries are highlighted in this issue. And finally, an example of the possible integrated international practice in patient education in the field of diabetes is described in this issue. Several conclusions are drawn concerning future development of communication in health care within the framework of patient education in Europe. PMID- 11390151 TI - A European perspective: common developments, differences and challenges in patient education. AB - In May 1999, a European meeting was held in Paris on the evolution and developments of patient education. The situation in different European countries and the factors influencing the development and improvement of patient education were discussed. Five categories of factors were identified: research and evidence based standards, the organization of care, training and methodological support, professional value and acknowledgement, funding and place of patient education in health policy. Several common orientations and priorities for the development of patient education have been presented, and are highlighted in this paper. PMID- 11390152 TI - Patient education in The Netherlands. AB - This article presents the development of patient education (PE) in The Netherlands from a historical perspective. A description is given of the first pioneering years from the 70s till the late 80s, in which early topics like the organization of PE, the orchestration of PE between different disciplines, the role of the social environment, the provision of PE in difficult patient groups and--most of all--the technical development of educational materials took the time and attention of the growing group of professionals that were engaged in patient education. Recent developments concern the legal aspects of PE, national policy, the role of health insurance, the position of patient organizations and the development of patient education in specific professional groups, e.g. general practitioners, nurses, physiotherapists, pharmacists, and dentists. There is no doubt that patient education has been professionalized considerably during the last decades. Nevertheless, new issues emerge and some old issues still need to be solved. The effective use of information material, the need for counseling as part of PE, and the relevance of coordination of care are longtime, but still actual problems in PE. More recent issues are the pressures on PE because of capacity restraints, the influence of the media and perhaps most of all: the apparent need for a patient-centered attitude and a more two-sided way of communication. Finally, the future policy topics in the Dutch patient education are discussed. PMID- 11390153 TI - Evolution not revolution? The struggle for the recognition and development of patient education in the UK. AB - In this paper, a review of patient education in the United Kingdom (UK) is presented. Some of the reasons for the development of patient education are identified and its relationship to other educational activities in health care settings is discussed. The gap between the theory and practice of UK patient education is then examined and emerging new practices are considered. The concluding section of the paper identifies a number of important questions to ask when considering the future development of patient education. These are relevant questions for patients, practitioners, researchers and politicians. PMID- 11390154 TI - Towards promotion, structuring and acknowledgement of patient education activities in France. AB - As patient education becomes more and more widespread in use among French health professionals, it is also becoming increasingly structured, but remains very heterogeneous depending on practitioners and institutions. For several years, this activity has been integrated into various professions. Training is becoming more frequent, and care providers show a certain willingness for common though on improvement and evaluation of their practices. At the same time, health institutions show an increasing interest in these activities, which they wish to promote. Significant improvements recently observed by them may finally lead to the professional and financial recognition expected by care providers. PMID- 11390155 TI - Implications of structural deficits for patient education in Germany. AB - Several changes in health politics and legal settings in recent years have affected the structure and practice of health promotion and patient education in Germany. The current legal background and its implications for patient education are discussed. Based on examples from four selected areas (cardiovascular diseases, diabetes mellitus, chronic pain, and asthma) the current practice of patient education in Germany is summarized. While many well-structured programs exist that are based on state-of-the-art guidelines, there is a lack of high quality research that documents the long-term effectiveness and cost effectiveness of such approaches. Structural problems and an insufficient number of highly trained personnel result in the fact that many patients do not have access to standardized programs. Persisting compliance problems indicate that there is still room for improvement of patient education interventions. As important for the future, necessary changes in the legal settings and possible implications for the education of the educators are discussed. PMID- 11390156 TI - Patient education in Belgium: evolution, policy and perspectives. AB - Patient education started in Belgium in the late 70s. Tuberculosis and diabetes management and care were the first topics addressed. In the two main regions of the country (Flemish and French), the development of patient education has been very different. The Belgian French Ministry of Health and regional hospital associations appointed a non-profit resource center, the "Center d'Education du Patient", in order to help hospital departments and health care teams start and improve patient education work and programs. University training programmes were created in the 80s, and an inter-hospital network organized to facilitate collaborations. Later, patient education hospital committees and coordinators were appointed, and professional organizations (patient education nurses and coordinators organization) were set up. In 1999, 98% of French speaking hospitals state that they have patient education programmes (three per hospital, on average), a remarkable growth from 7% in 1983. Belgium has joined the WHO health promoting hospitals project in 1996. In private practice, due to the "payment on service" system that does not allow means for patient education work, patient education is still rare. In the Flemish region, patient education programmes exist in some hospitals, on private initiatives, without support from the Ministry of Health, nor from the health promotion agency. On the conceptual side, programmes have shifted from a "patient instruction" perspective focusing on the biomedical aspects of health and disease, and professional expertize and needs assessment, to "patient participation" dealing with biopsychosocial health and disease. Lay and subjective needs and "life projects" are more and more taken as a basis for patient counseling and therapeutic education. With the renewed involvement of the government in patients rights, and the possibility to start funding patient education as any other care work, new developments of patient education are expected in the next years. PMID- 11390157 TI - Patient education in Finland. AB - No mandated policies, practices, and procedures of patient education or patient health education exist in Finland but the efforts have been local in nature. Traditionally, primary and secondary health education as a part of maternal health care and during dental appointments have been known to exist. Hospital patient education efforts which may have resulted from past public health problems or legislative endeavors do exist in many hospital units but official hospital policies or strategies still do not. Patient education is often seen as health education. Voluntary agencies tend to emphasize practical issues rather than strategic planning in patient education. Finland complies with the WHO health strategies of 2015, and as a part of the Finnish strategies, it is considered important that large groups of people participate. A recent movement in health care to plan and implement shared decision-making activities (Seamless Macro Project, Path-project) is still short-term in nature. Practical actions are guided by professional skills, ethical practices, professional norms, and habits of the land, and of the three laws (public health law, occupational health law, patient status and rights law) guiding health education activities, the third one is specifically directed to patient education with an intent that when a treatment is not acceptable, another acceptable one must be found. Empowerment is a term seen in many contexts. PMID- 11390158 TI - Patient education--new trends in Sweden. AB - Patient education has a long history in Sweden and the field of diabetes care has been in the forefront of the development since the last 20 years. It is now well recognised that patients with chronic diseases must know how to handle their situation and that the long-term outcomes very much depend on how successful they are in their endeavour. More recently, patient education was also provided outside the medical services. Group education and a new role of the educator as a tutor and facilitator has emerged. Education is imparted more and more in the form of experiential learning. PMID- 11390159 TI - Current perspectives of therapeutic patient education in Italy. AB - Therapeutic patient education is a well-defined branch of health education aimed at patient empowerment. It consists of helping the patient to understand his own disease and its treatment, actively collaborating to its fulfillment and to take care of his own health status in order to maintain and improve his life quality. The correct implementation of both communication and therapeutic patient education involves the mastery of specific professional skills by healthcare personnel. In Italy, institutional therapeutic patient education is delivered mainly to diabetic patients. However, other activities and projects aimed at therapeutic education of chronic patients are gradually appearing. An overview of current situation and perspective of therapeutic patient education practice in Italy are presented. PMID- 11390160 TI - Patient education in Switzerland: from diabetes to chronic diseases. AB - The Division of Therapeutic Education for Chronic Diseases at the University Hospital of Geneva has been playing an important role in the field of therapeutic patient education for more than 25 years. More than 16,000 patients have been hospitalised and an excess of 75,000 h have been spent with a rather novel interdisciplinary approach involving doctors, nurses, dieticians, psychologists, podiatrists and pedagogues. For the past 12 years, our division has held over 50 seminars of 1-week postgraduate training attended by over 3000 participants coming from more than 60 countries worldwide. In 1998, the faculty of medicine at the University of Geneva implemented a 3-year curriculum on therapeutic patient education leading to a postgraduate university diploma. In 1983, the WHO designated the Swiss teaching division as a WHO Collaborating Center for reference and research in diabetes education. In 1998, a WHO-Euro Working Group Report entitled "Therapeutic Patient Education. Continuing education programmes for health care providers in the field of prevention of chronic diseases" was published. PMID- 11390161 TI - Patient education in Hungary. AB - Patient education is a highly context dependent activity. The Hungarian scene is conditioned by three well articulated aspects. First, the well developed and still cherished roots of dissemination of knowledge; second, the traditional sensitivity of ethical issues in health related, including death and dying issues; third, the present fast socio-political changes in this country. The health promoting hospital movement's widespread activities as well as behavioral sciences, mental health and health psychology aspects of patient education are also reviewed. The theoretical as well as skills aspects of communication and systematic attempts to increase the effectiveness of interactions are also presented. PMID- 11390162 TI - Patient education in Estonia. AB - Following the developments in Health Promotion in Estonian Health Care during the past 5-6 years, it is possible to observe remarkable progress in this field. The fundamental achievement is the shifting of a patient into the attention centre of the health care system. The patient-oriented approach means: to increase the involvement of the patient and his/her family in valuing and maintaining their health and modelling healthy behaviour; coping with chronic disease in relation to everyday activities; patient satisfaction; psycho-social support and counselling. The patient education is a multiprofessional teamwork and interdisciplinary co-operation that should be carried out in two levels mainly: primary health care and hospital levels. The key-persons in patient education are family doctors, nurses and trained hospital staff. It is essential for the development of health promotion in hospitals to incorporate the idea of health promotion as an integrated part of everyday work in addition to the traditional curative function. PMID- 11390163 TI - Current perspectives on patient education in the US. AB - Patient education has evolved from its medically-dominated and narrow origin in patient teaching to support of patient empowerment in interpersonal, organizational, and policy domains relevant to health. This essay reflects on both the historical and contemporary context of patient education in the US and explores implications of the empowerment movement on new initiatives and directions in patient education. By using diabetes education as an exemplar, innovations in patient activation and empowerment are explored and future directions and challenges to the field are considered. PMID- 11390164 TI - The diabetes education study group and its activities to improve the education of people with diabetes in Europe. AB - In this article the activities of the diabetes education study group (DESG) are presented. It is an important example of the coordination of the European patient education in the field of diabetes. Given the therapeutic role of patient education, doctors must be involved in the entire process, as members of the multi-professional team, who carry the responsibility for the planning and implementation of the educational process. Based on these assumptions, the DESG concentrated on the following activities: workshops, congresses, teaching letters, a 5-min education kit, a web-site, and a basic curriculum in therapeutic patient education. The future of therapeutic patient education in the field of diabetes is discussed. PMID- 11390165 TI - Glyburide and fetal safety; transplacental pharmacokinetic considerations. AB - Oral hypoglycemics have been avoided in pregnancy due to their potential to cause fetal hyperinsulinemia/hypoglycemia. A recent human study has shown glyburide to minimally cross the placenta, allowing a safe new treatment for gestational diabetes. The mechanisms for the minimal placental passage of this small molecule are not clear. In this presentation, the role of pKa, molecular weight, lipid solubility, and protein binding is considered. Out of these physical and pharmacologic characteristics, the very extensive plasma protein binding and short elimination half-life of glyburide appear to be major determinants of its minimal transplacental transfer. PMID- 11390166 TI - Tampons, dioxins, and endometriosis. AB - Concern has been expressed that rayon tampons contain dioxins as a result of chlorine bleaching and, further, that the dioxins in tampons may increase the risk of endometriosis. Rayon tampons do not contain 2,3,7,8-tetrachlorodibenzo-p dioxin, the chemical commonly meant when the generic term "dioxin" is used. In addition, rayon tampons contain only trivial amounts of dioxin-like environmental contaminants, similar to the amounts contained in unbleached cotton tampons. The amount of dioxin-like material that is theoretically available from tampons is at least six orders of magnitude lower than estimated daily food exposure levels to these contaminants. The evidence for a causal relationship between environmental exposure to dioxins and endometriosis is inconsistent. Prediction of the effective dioxin dose based on the most suggestive of the primate studies on endometriosis does not raise concerns about typical human food exposures to these compounds, let alone the considerably lower levels that could be present in tampons. PMID- 11390167 TI - Lack of embryotoxicity of homocysteine thiolactone in mouse embryos in vitro. AB - Recent work from humans and chick embryos has suggested that homocysteine may play a role in producing neural tube defects (NTDs). In an effort to determine if homocysteine is able to produce NTDs in mammalian embryos, mouse embryos were explanted on GD 8 and cultured for 44 h. When either homocysteine or homocysteine thiolactone was added to the culture medium, treated embryos developed as well as controls and had closed neural tubes. Homocysteine thiolactone was also microinjected into the amniotic sac of mouse embryos. Again, development proceeded normally with no significant increase in the number of embryos with open neural tubes at the end of the culture period. HPLC analysis of embryonic thiols 24 h after microinjection revealed a significant increase in embryonic cystathionine levels. These data suggest that homocysteine does not produce NTDs in mouse embryos cultured in vitro and that early organogenesis-stage embryos are able to metabolize homocysteine. PMID- 11390168 TI - Cell response endpoints enhance sensitivity of the immature mouse uterotropic assay. AB - Outbred immature CD-1 mice were subcutaneously (s.c.) injected once on postnatal day 17 or on postnatal days 17, 18, and 19 with 17beta-estradiol, diethylstilbestrol, tamoxifen, 4-hydroxytamoxifen, methoxychlor, the methoxychlor metabolite HPTE, nonylphenol, o,p'-DDT, endosulfan, or kepone over a wide dose range (0.1 to 1,000,000 microg/kg). On the day following the last injection, uterine weight/body weight ratios were determined and uterine tissues processed for histologic examination. All compounds except endosulfan and kepone increased uterine wet weight compared to vehicle controls; however, the dose response curve and magnitude of response varied depending on the compound. Choosing the maximum wet weight dose for each compound, uterine tissue was evaluated for epithelial cell height, epithelial and stromal cell proliferation, endometrial gland number, and induction of estrogen-inducible proteins lactoferrin and complement C3. All compounds elicited estrogen-responsive changes in these endpoints that were individually more sensitive than uterine weight alone. We conclude that these endpoints enhance the sensitivity of the uterotropic bioassay. PMID- 11390170 TI - Effects of monobutyl phthalate on reproductive function in pregnant and pseudopregnant rats. AB - The effects of monobutyl phthalate (MBuP) on reproductive function were determined in pregnant and pseudopregnant rats. Rats were given MBuP by gastric intubation at 250, 500, 750, or 1000 mg/kg on days 0 to 8 of pregnancy and pregnancy outcome was determined on day 20 of pregnancy. The effects of MBuP on the uterine function, as a cause of early embryonic loss, were also determined in pseudopregnant rats, with an induced decidual cell response. The same doses of MBuP were given to pseudopregnant rats on days 0 to 8 of pseudopregnancy and the uterine weight on day 9 served as an index of uterine decidualization. MBuP at 1000 mg/kg caused significant increases in the incidences of preimplantation loss in females successfully mated and of postimplantation loss in females having implantations. Uterine decidualization in pseudopregnant rats was significantly decreased at 1000 mg/kg. These findings suggest that early embryonic loss due to MBuP is mediated, at least in part, via suppression of uterine decidualization, an impairment of uterine function. PMID- 11390169 TI - A mechanistic assessment of 1,3-butadiene diepoxide-induced inhibition of uterine deciduoma proliferation in pseudopregnant rats. AB - Butadiene diepoxide (BDE), a reactive metabolite of 1,3-butadiene that is an important industrial chemical used in synthetic rubber production causes a dose dependent inhibition of deciduoma development in pseudopregnant Sprague-Dawley rats. This study used 4 daily i.p. BDE doses of 0.20, 0.25, 0.30, 0.35, or 0.40 to characterize mechanisms that may be responsible for the antideciduoma effect. Pseudopregnant rats were treated either before (pseudopregnancy [PPG] days 1-4) or after (PPG days 5-9) deciduoma induction by endometrial trauma with a blunt needle. Animals were killed on PPG day 9 and evaluated for serum progesterone and endometrial protein and DNA. RT-PCR was used to measure message for estrogen receptor (ER) alpha and pituitary adenylate cyclase-activating polypeptide (PACAP). Substrate zymography and Western blotting were used respectively to measure matrix metalloproteinase (MMP)-9 and inducible nitric oxide synthase. The antideciduoma effects of BDE were associated with decreases in endometrial weight, protein, and DNA, with decreases in serum progesterone, and with decreases in PACAP message and MMP-9. A reduction in NOS was identified at the highest dose of BDE. Message for estrogen receptor (ER) alpha was not affected at any dose. We conclude that the reduction in decidual proliferation was direct and appeared to be associated with either 1) a decrease in the effectiveness of the deciduogenic stimulation and/or a weakened endometrial sensitivity to the stimulus; or 2) an effect on deciduoma development. Molecular mechanisms that apparently contributed to BDE inhibition of decidual metabolism included the synthesis of protein and DNA involved in decidual growth, the synthesis and activation of a matrix metalloproteinase for degradation of the extracellular matrix that is essential for tissue remodeling during deciduoma development, and the nitric oxide/nitric oxide synthase and pituitary adenylate cyclase-activating peptide systems that are involved in promoting vasodilation and increased vascular permeability to enhance the availability of substrates for maximal deciduoma growth. The ovotoxicity of BDE, which has previously been established, may indirectly affect decidual proliferation by reducing progesterone, the preeminent endocrine regulator of deciduoma development. The findings also suggest that BDE may possess no estrogenic action since it was associated with endometrial weight loss and unaltered levels of the estrogen receptor alpha mRNA expression. PMID- 11390171 TI - The effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on weight gain and hepatic ethoxyresorufin-o-deethylase (EROD) induction vary with ovarian hormonal status in the immature gonadotropin-primed rat model. AB - Immature female rats received 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) during an induced proestrus or diestrus. The inhibitory effect of TCDD on acute weight gain and the induction of hepatic ethoxyresorufin-o-deethylase (EROD) activity by TCDD were greatest during proestrus. In a second experiment, ovariectomized rats received estradiol cypionate (ECP) or progesterone followed by TCDD. TCDD and estradiol each alone significantly inhibited weight gain. Progesterone potentiated the effects of TCDD on weight gain. The highest dose of ECP was associated with greater induction of hepatic EROD activity by TCDD than seen with TCDD alone. Estradiol modulates the induction of hepatic EROD activity by TCDD. Differential effects of TCDD on acute weight gain during proestrus vs. diestrus in this model do not mimic changes induced by estrogen alone. Hepatic responses to TCDD may vary according to phase of the female reproductive cycle. PMID- 11390172 TI - Impaired ovulation by 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) in immature rats treated with equine chorionic gonadotropin. AB - Several studies have established that 2,3,7,8 tetrachloro-p-dioxin (TCDD) blocks ovulation. The main purpose of this study was to determine if induced ovulation was delayed temporarily by TCDD. The ovulation model used was that of the gonadotropin-primed intact or hypophysectomized rat. Immature intact female Sprague-Dawley rats (IIR) were given 32 microg TCDD/kg by gavage on day 24 of age. The next day equine chorionic gonadotropin (eCG) (5 IU) was injected sc to stimulate follicular development. The number of ova in the oviducts, the ovulation rate, and steroid concentrations were determined at 72, 96, 120, and 144 h after eCG. Immature female Sprague-Dawley rats (IHR) were hypophysectomized on day 23 of age. On day 26, the IHR were given 20 microg TCDD/kg by gavage. The next day eCG (10 IU) was injected sc to stimulate follicle development and at 52 h after eCG, 10 IU human chorionic gonadotropin (hCG) was given to induce ovulation. The same parameters as in IIR were determined in IHR at 72, 96, and 120 h after eCG. TCDD decreased body and ovarian weight gains in both IIR and IHR. In IIR, TCDD delayed ovulation by 24 to 48 h reducing the number of ova shed as well as the number of animals ovulating at 72 and 96 h after eCG. In IHR, however, TCDD reduced only the number of ova shed but caused no delay in ovulation. The IIR treated with TCDD had low levels of progesterone (P4) at 72 and 96 h after eCG but high levels of estradiol (E2) at the same time points. This sustained high level of E2 production coincided with a transient decrease in serum concentrations of androstenedione (A4). The alteration of steroid hormones by TCDD was restored to normal by 48 h after ovulation in IIR. Serum P4 concentration was not altered by TCDD in IHR at 72 h after eCG but was decreased thereafter. The delay in ovulation induced by TCDD in IIR indicates the disruption of the hypothalamus-pituitary-ovary axis during proestrus. The decrease in number of ova shed in IHR induced by exogenous gonadotropins indicates an additional direct ovarian effect of TCDD in blocking ovulation. PMID- 11390173 TI - Effects of gestational and lactational exposure to Aroclor 1242 on sperm quality and in vitro fertility in early adult and middle-aged mice. AB - The objective of this study was to examine the effects of gestational and lactational exposure to Aroclor 1242 (0, 10, 25, 50, and 100 mg/kg-bw) on male fertility. Doses were administered to C57BL6 female mice orally every two days from two weeks before mating, during mating, and through gestation until postnatal day 21. Male B6D2F1 offspring were examined for anogenital distance, organ development, epididymal sperm count, sperm motility, and in vitro fertility at 16 and 45 weeks of age. Stomach samples of pups nursing from PCB-treated mothers in the 50 mg/kg dose group were analyzed for PCBs and chlorobiphenylols by high resolution gas chromatography coupled with low resolution mass spectrometry. It was estimated that the nursing pups were exposed to 0.2, 0.6, 1.2, and 2.4 mg/kg/day total PCBs in the 10, 25, 50, and 100 mg/kg dose groups, respectively. This exposure level approaches the maximum FDA recommended levels for PCBs in food and breast milk. The composition of the PCBs in the stomach samples was different from the parent mixture, as there was a higher proportion of heavily chlorinated congeners, as well as chlorobiphenylols. Anogenital distance at weaning, and liver, thymus, and testes weight at 16 and 45 weeks of age were not affected by PCB exposure. Epididymal sperm velocity and linearity were significantly increased in the 25 mg/kg dose group at 16 weeks of age. Sperm count was increased by 36% in this dose group (P = 0.06). By 45 weeks of age, average sperm count in this dose group was similar to that of controls. With the exception of the 50 mg/kg dose group at 16 weeks of age, sperm fertilizing ability in vitro was significantly decreased in all PCB-exposed groups at 16 and 45 weeks of age. These results suggest that fertility in the adult mouse is susceptible to developmental exposure to Aroclor 1242 and is independent of testis weight or epididymal sperm count. PMID- 11390174 TI - Reproductive effects of nonylphenol in rats after gavage administration: a two generation study. AB - The potential reproductive toxicity of nonylphenol (NP) was assessed in a two generation reproductive toxicity study. Groups of 25 male and female Crj:CD (SD) IGS rats were given NP by gavage at levels of 2, 10, or 50 mg/kg, and 25 males and females were given corn oil as controls. No adverse changes in clinical signs were observed in any rats throughout the study. Significant increases in the liver, kidney and pituitary gland weights in males, and decreases in thymus weight in males and in ovary weight in females were observed in the 50 mg/kg group. NP did not affect sperm characteristics or the estrous cycle at any dose administered. A significant increase in the TSH level was observed in males in the 50 mg/kg group. No adverse effects of NP on reproduction were found. At necropsy, no treatment-related alterations were observed in any organs including the reproductive tissues in any group. Histopathologic changes were found in the liver of male and female rats and kidneys of males in the 50 mg/kg group. The viability of offspring from postnatal day 0 to 4 in the 50 mg/kg group was reduced as compared with that in the controls, although growth was not affected by NP administration. On postnatal day 22, an increase in the serum FSH level and decrease in T(3) level for males, and decreases in LH and TSH levels and an increase in T(3) levels for females were observed in the 50 mg/kg group. NP did not affect the timing of preputial separation, while vaginal opening was accelerated in the 50 mg/kg group. No adverse changes were found in behavior or learning in the offspring of NP-treated groups. There were no treatment-related changes in any reproductive parameter, including estrous cycle, mating, fertility, delivery, and lactation, except for significant decreases in the numbers of implantation sites and live pups, and a significant decrease in ovary weight in the 50 mg/kg group. Kidney and liver weight were increased in males in the 50 mg/kg group. Histopathologic examination revealed changes in the liver of males and females of the 50 mg/kg group. No treatment-related changes were observed in the sperm characteristics. Hormone data should be interpreted cautiously until the findings are repeated and confirmed by further studies. These results of NP suggested that the no observed adverse effect level (NOAEL) on reproductive capacity is 50 mg/kg/day or greater in parent animals, and 10 mg/kg/day in the next generation under the present experimental condition. PMID- 11390175 TI - Actions of the endocrine disruptor methoxychlor and its estrogenic metabolite on in vitro embryonic rat seminiferous cord formation and perinatal testis growth. AB - The current study examines the actions of methoxychlor and its estrogenic metabolite, 2, 2-bis-(p-hydroxyphenyl)-1, 1, 1-trichloroethane (HPTE), on seminiferous cord formation and growth of the developing rat testis. The developing testis in the embryonic and early postnatal period is likely more sensitive to hormonally active agents than at later stages of development. Embryonic day 13 (E13) testis organ cultures were treated with either 0.2, 2, or 20 microM methoxychlor or 1, 3, 6, 15, 30, or 60 microM HPTE to examine effects on cord formation. No concentration of methoxychlor completely inhibited cord formation. However, cord formation was abnormal with the presence of a reduced number of cords and appearance of "swollen" cords at the 2 and 20 microM concentrations of methoxychlor. The swollen cords were due to an increase in the number of cells in a cord cross section and reduction of interstitial cell numbers between cords. Treatment of embryonic day 13 (E13) testes with HPTE caused abnormal cord formation at the 3 microM and 6 microM concentrations, and completely inhibited cord formation at the 15, 30, and 60 microM concentrations. In addition to the estrogenic metabolite HTPE, methoxychlor can also be metabolized into anti-androgenic compounds. Therefore, to determine the spectrum of potential actions of methoxychlor on testis development, different concentrations of estradiol, testosterone, and an anti-androgen (flutamide) were utilized to determine their effects on E13 testis organ culture morphology. Estradiol (1 microM) and flutamide (0.1microM) both inhibited seminiferous cord formation in E13 testis organ cultures. Therefore, methoxychlor may be acting through the androgen and/or estrogen receptors to elicit its actions on seminiferous cord formation. Reverse transcription polymerase chain reaction (PCR) (RT-PCR) confirmed the presence of estrogen receptor alpha (ERalpha) mRNA from embryonic day 14 (E14) through postnatal day 5 (P5) while estrogen receptor beta (ERbeta) mRNA did not appear until approximately E16 of testis development. Androgen receptor (AR) expression was present from E14 through P5 of testis development, but at apparently reduced levels at E14 and E16. Immunohistochemical analysis localized ERalpha to the cells of the seminiferous cords at E14 though P5 while ERbeta was present in cells of the interstitium at E16 and P0. Androgen receptor was localized to germ and interstitial cells. The effects of methoxychlor, HPTE, estradiol, and testosterone on cell growth of perinatal testes was determined with a thymidine incorporation assay in postnatal day zero (P0) testis cell cultures. Methoxychlor (0.002, 0.02, and 0.2 microM) and HPTE (2 and 20 microM) stimulated thymidine incorporation in P0 testis cell cultures in a similar manner to estradiol (0.01, 0.1, and 1 microM). In addition, testosterone (0.1 microM) also stimulated thymidine incorporation in P0 testis cultures. Observations suggest that methoxychlor and its metabolite HPTE can alter normal embryonic testis development and growth. The actions of methoxychlor and HPTE are likely mediated in part through the steroid receptors confirmed to be present in the developing testis. PMID- 11390176 TI - Embryotoxicity of meglumine antimoniate in the rat. AB - Meglumine antimoniate (MA) is a pentavalent antimonial (Sb(V)) drug used to treat leishmaniasis. Despite the fact that Sb(V) organic compounds have been used in clinical practice for more than 50 years, information on their safety during pregnancy is still scanty. This study was undertaken to evaluate the embryo/fetotoxicity of MA in the rat. Wistar rats were treated subcutaneously (s.c.) with MA (300 mg Sb(V)/kg body wt/day) on days 6 through 15 of pregnancy or with a higher dose (3 x 300 mg Sb(V)/kg body wt) on day 11 only. A control group treated with saline on days 6 through 15 and an untreated control group were evaluated as well. Cesarean sections were performed on day 21. No maternal toxicity and no reduction of fetal weight were noted in the groups treated with MA. The repeated administration of MA (days 6 through 15), but not the acute treatment (day 11), enhanced embryolethality. Treatment with MA on days 6 through 15 also caused a higher incidence of an atlas bone anomaly that occurs spontaneously at very low frequencies in our rat strain. These findings indicated that repeated administration of MA was embryolethal and teratogenic in rats. PMID- 11390177 TI - The mammalian testis accumulates lower levels of organochlorine chemicals compared with other tissues. AB - Tissues were obtained from three separate experiments in order to quantify the tissue distribution of organochlorine chemicals that are thought to be potential reproductive toxicants in males: 1) Sprague Dawley rats received 1 microCi of 14C Aldrin or 14C-Dieldrin (20.6 microCi/micromole) i.p. once a week for three weeks. One week and four weeks after the last injection, tissues were harvested and stored at -80 degrees C. Tissue 14C levels were quantified by scintillation spectrometry. 2) Cis- or trans-nonachlor (0, 0.25, 2.5, 25 mg/kg body weight) were administered daily in corn oil to male rats by gavage for 28 days. Tissues were harvested and frozen at -80 degrees C on the 29th day. Organochlorine residues were extracted and quantified by gas chromatography with electron capture detection. 3) Technical grade toxaphene (0, 0.1, 0.4 or 0.8 mg/kg body weight) was ingested daily by female cynomolgus monkeys of reproductive age for 18 months prior to being mated with control males. Dosing continued during pregnancy and lactation. Their infants received toxaphene via breast milk, and upon weaning, they ingested the same dose as their mothers for 48 to 49 weeks until, at 77 to 80 weeks of age, tissues were harvested and stored at -80 degrees C. Organochlorine residues were extracted and quantified as previously stated. In all three experiments, organochlorine residues in the testis were lower than in most of the other reproductive tract and nonreproductive tract tissues we examined. For example, testicular aldrin and dieldrin levels were <5% the epididymal content; testicular cis- and trans-nonachlor were <25% the epididymal content and, testicular toxaphene levels were <15% of the epididymal content. The reasons for the low degree of accumulation by the testis in comparison with other tissues are unknown. However, the lower testicular content may afford germ cells some protection from the potentially toxic effects of these chemicals. PMID- 11390178 TI - Isoprostane levels in lipids extracted from atherosclerotic arteries of nonhuman primates. AB - Nonhuman primates used in these studies had been fed for 5 years diets enriched with cholesterol and one of three classes of fatty acids: saturated, monounsaturated, or polyunsaturated fatty acids. Atherosclerotic iliac artery lipid extracts were quantitatively analyzed for cholesterol, cholesteryl esters, fatty acid composition, and a marker of lipid oxidation, the F(2)-isoprostanes. There was no significant difference in the mean accumulation of F(2)-isoprostanes among the different diet groups. To account for the small, individual variation in the arachidonate concentration the F(2)-isoprostane mass from each sample was normalized by dividing by arachidonate mass: F(2)-isoprostane mass/(mass arachidonate). At lower levels of cholesterol accumulation, the F(2)-isoprostane mass/(mass arachidonate) ratio was greater in lipids from POLY arteries compared to SAT arteries, but the reverse was true at high levels of cholesterol. F(2) isoprostane/(mass arachidonate) increased with mole fraction linoleate for the SAT group, but decreased for the POLY group. In summary, these studies demonstrated that there is no simple explanation of how F(2)-isoprostane accumulation did not depend on the concentration of oxidizable lipids that promote free-radical lipid oxidation. PMID- 11390179 TI - Methylmercury and H(2)O(2) provoke lysosomal damage in human astrocytoma D384 cells followed by apoptosis. AB - Methylmercury (MeHg) is a neurotoxic agent acting via diverse mechanisms, including oxidative stress. MeHg also induces astrocytic dysfunction, which can contribute to neuronal damage. The cellular effects of MeHg were investigated in human astrocytoma D384 cells, with special reference to the induction of oxidative-stress-related events. Lysosomal rupture was detected after short MeHg exposure (1 microM, 1 h) in cells maintaining plasma membrane integrity. Disruption of lysosomes was also observed after hydrogen peroxide (H(2)O(2)) exposure (100 microM, 1 h), supporting the hypothesis that lysosomal membranes represent a possible target of agents causing oxidative stress. The lysosomal alterations induced by MeHg and H(2)O(2) preceded a decrease of the mitochondrial potential. At later time points, both toxic agents caused the appearance of cells with apoptotic morphology, chromatin condensation, and regular DNA fragmentation. However, MeHg and H(2)O(2) stimulated divergent pathways, with caspases being activated only by H(2)O(2). The caspase inhibitor z-VAD-fmk did not prevent DNA fragmentation induced by H(2)O(2), suggesting that the formation of high molecular-weight DNA fragments was caspase independent with both MeHg and H(2)O(2). The data point to the possibility that lysosomal hydrolytic enzymes act as executor factors in D384 cell death induced by oxidative stress. PMID- 11390180 TI - Trimetazidine protects low-density lipoproteins from oxidation and cultured cells exposed to H(2)O(2) from DNA damage. AB - Trimetazidine is a well-established anti-ischemic drug, which has been used for long time in the treatment of pathological conditions related with the generation of reactive oxygen species. However, although extensively studied, its molecular mode of action remains largely unknown. In the present study, the ability of trimetazidine to protect low-density lipoproteins (LDL) from oxidation and cultured cells from H(2)O(2)-induced DNA damage was investigated. Trimetazidine, tested at concentrations 0.02 to 2.20 mM, was shown to offer significant protection to LDL exposed to three different oxidizing systems, namely copper, Fe/ascorbate, and met-myoglobin/H(2)O(2). The oxidizability of LDL was estimated by measuring, (i) the lag period, (ii) the maximal rate of conjugated diene formation, (iii) the total amount of conjugated dienes formed, (iv) the electrophoretic migration of LDL protein in agarose gels (REM), and (v) the inactivation of the enzyme PAF-acetylhydrolase present in LDL. In addition, the presence of trimetazidine decreased considerably the DNA damage in H(2)O(2) exposed Jurkat cells in culture. H(2)O(2) was continuously generated by the action of glucose oxidase at a rate of 11.8 +/- 1.5 microM per min (60 ng enzyme per 100 microl), and DNA damage was assessed by the single cell gel electrophoresis assay (also called comet assay). The protection offered by trimetazidine in this system (about 30% at best) was transient, indicating modification of this agent during its action. These results indicate that trimetazidine can modulate the action of oxidizing agents in different systems. Although its mode of action is not clarified, the possibility that it acts as a lipid barrier permeable transition metal chelator is considered. PMID- 11390181 TI - Relationship between posttranslational modification of transaldolase and catalase deficiency in UV-sensitive repair-deficient xeroderma pigmentosum fibroblasts and SV40-transformed human cells. AB - Xeroderma Pigmentosum (XP) is a rare recessively inherited human disease associated with a hypersensitivity to ultraviolet radiation. The ultraviolet component of sunlight can initiate and promote the formation of cutaneous tumors as seen in the skin cancer-prone XP patients. Previously, we have found that the low activity of the NADPH-dependent antioxydant enzyme, catalase, which we have observed in XP diploid fibroblasts and SV40-tranformed cells, could be restored by the addition of NADPH. Here we have analyzed transaldolase, which regulates NADPH levels produced by the pentose phosphate pathway in order to examine how it influences the catalase activity regulated in XP and SV40-transformed cells. We find that transaldolase activity is high in XP and SV40-transformed human fibroblasts, whereas transaldolase transcription is unchanged, suggesting that modification of transaldolase activity is due to a posttranslational modification of the protein. Two-dimensional electrophoresis analysis has allowed us to identify a complex set of transaldolase isoforms and to postulate that the phosphorylation of specific isoforms could be correlated with the different enzymatic activities seen. Our results show that high transaldolase activity corresponds to a low catalase activity in SV40-transformed cells and in fibroblasts from XP patients who have a high predisposition to develop skin cancer. PMID- 11390182 TI - The role of the DMPO-hydrated electron spin adduct in DMPO-*OH spin trapping. AB - Time-resolved in situ radiolysis ESR (electron spin resonance, equivalently EPR, electron paramagnetic resonance) studies have shown that the scavenging of radiolytically produced hydroxyl radical in nitrous oxide-saturated aqueous solutions containing 2 mM DMPO is essentially quantitative (94% of the theoretical yield) at 100 micros after the electron pulse [1]. This result appeared to conflict with earlier results using continuous cobalt-60 gamma radiolysis and hydrogen peroxide photolysis, where factors of 35 and 33% were obtained, respectively [2,3]. To investigate this discrepancy, nitrogen-saturated aqueous solutions containing 15 mM DMPO were cobalt-60 gamma irradiated (dose rate = 223 Gy/min) for periods of 0.25-6 min, and ESR absorption spectra were observed approximately 30 s after irradiation. A rapid, pseudo-first-order termination reaction of the protonated DMPO-hydrated electron adduct (DMPO-H) with DMPO-OH was observed for the first time. The rate constant for the reaction of DMPO-H with DMPO-OH is 2.44 x 10(2) (+/- 2.2 x 10(1)) M(-1) s(-1). In low-dose radiolysis experiments, this reaction lowers the observed yield of DMPO-OH to 44% of the radiation-chemical OH radical yield (G = 2.8), in good agreement with the earlier results [2,3]. In the absence of the DMPO-H radical, the DMPO-OH exhibits second-order radical termination kinetics, 2k(T) = 22 (+/- 2) M(-1) s(-1) at initial DMPO-OH concentrations > or = 13 microM, with first-order termination kinetics observed at lower concentrations, in agreement with earlier literature reports [4]. PMID- 11390183 TI - Differential inhibition by alpha- and beta-tocopherol of human erythroleukemia cell adhesion: role of integrins. AB - The effect of alpha- and beta-tocopherol on human erythroleukemia cell (HEL) adhesion induced by phorbol 12-myristate 13-acetate (PMA) has been studied. Adhesion induced by PMA stimulation was prevented by 44.5% by physiological concentrations of alpha-tocopherol. Under the same experimental conditions, beta tocopherol, an analogue of alpha-tocopherol, produced 11% inhibition of adhesion. Cell response gradually increased from 0 to 24 h of alpha-tocopherol treatment. Only a slight time dependency of beta-tocopherol inhibition was observed. Another human erythroleukemia cell line (K562) and the human monocyte tumor cell line U937 showed 5.0 and 11.2% inhibition, respectively. Similar to alpha-tocopherol, the protein kinase C inhibitor, Calphostin C, and the MAPK inhibitor, PD98059, prevented PMA-induced cell adhesion. An inhibition of ERK-1 phosphorylation was observed for alpha-tocopherol only in HEL, implying that MAP kinase pathway is involved in this cell line. Fluorescence-activated cell sorting (FACS), by using various integrin-specific monoclonal antibodies, has shown that alpha (1-6), beta1, and alphav integrins are less expressed at the cell surface after alpha tocopherol treatment. Beta-tocopherol treatment was less effective. PMID- 11390184 TI - Free radical scavenging behavior of folic acid: evidence for possible antioxidant activity. AB - The free radical scavenging properties and possible antioxidant activity of folic acid are reported. Pulse radiolysis technique is employed to study the one electron oxidation of folic acid in homogeneous aqueous solution. The radicals used for this study are CCl(3)O(2)(*), N(3)(*), SO(4)(*-), Br(2)(*-), *OH, and O(*-). All these radicals react with folic acid under ambient condition at an almost diffusion-controlled rate producing two types of transients. The first transient absorption maximum is around 430 nm, which decays, and a simultaneous growth at around 390 nm is observed. Considering the chemical structure of folic acid, the absorption maximum at 430 nm has been assigned to a phenoxyl radical. The latter one is proposed to be a delocalized molecular radical. A permanent product has been observed in the oxidation of folic acid with CCl(3)O(2)(*) and N(3)(*) radicals, with a broad absorption band around 370-400 nm. The bimolecular rate constants for all the radical-induced oxidation reactions of folic acid have been measured. Folic acid is seen to scavenge these radicals very efficiently. In the reaction of thiyl radicals with folic acid, it has been observed that folic acid can not only scavenge thiyl radicals but can also repair these thiols at physiological pH. While carrying out the lipid peroxidation study, in spite of the fact that folic acid is considerably soluble in water, we observed a significant inhibition property in microsomal lipid peroxidation. A suitable mechanism for oxidation of folic acid and repair of thiyl radicals by folic acid has been proposed. PMID- 11390185 TI - The role of glutathione in the toxicity of smoke condensates from cigarettes that burn or heat tobacco. AB - Inhalation of cigarette smoke aerosol via active smoking is associated with the development of pulmonary inflammation. The cytotoxic potential of cigarette smoke has been hypothetically related to development of pulmonary inflammation since the release of intracellular contents from dead and dying cells has been reported to induce inflammatory foci. In this study, cigarette smoke condensates (CSCs) were prepared from Kentucky 1R4F reference cigarettes and cigarettes that primarily heat tobacco (Eclipse). The two CSCs were then compared for their ability to induce killing in human-hamster A(L) hybrid cells. CSCs prepared from Eclipse were much less cytotoxic than those prepared from reference cigarettes. At 60 microg CSC/ml culture medium, survival for CSC from Eclipse cigarettes was approximately 70% compared with 1% for CSC from burned K1R4F cigarettes. The observed reduction in CSC-Eclipse cytotoxicity toward these mammalian cells is consistent with the previously published observation of a 30% decline in pulmonary white cell count and 40% reduction in visual bronchitis index in human smokers who switched to Eclipse for 2 months. Results with N-acetylcysteine and buthionine-S-R-sulfoximine indicate that glutathione markedly reduces the cytoxicity of both CSCs. PMID- 11390186 TI - Oxidative DNA damage and cytotoxicity induced by copper-stimulated redox cycling of salsolinol, a neurotoxic tetrahydroisoquinoline alkaloid. AB - A series of neurotoxic tetrahydroisoquinoline alkaloids has been detected in certain regions of mammalian brains. One such dopaminergic tetrahydroisoquinoline neurotoxin is salsolinol (SAL), which is suspected of being associated with the etiology of Parkinson's disease and neuropathology of chronic alcoholism. In the present study, we found that SAL in combination with Cu(II) induced strand scission in pBR322 and phiX174 supercoiled DNA, which was inhibited by the copper chelator, reactive oxygen species (ROS) scavengers, reduced glutathione, and catalase. SAL in the presence of Cu(II) caused hydroxylation of salicylic acid to produce 2,3- and 2,5-dihydroxybenzoic acids. Reaction of calf thymus DNA with SAL plus Cu(II) resulted in substantial oxidative DNA damage as determined by 8 hydroxydeoxyguanosine (8-OH-dG) formation. Blockade of the dihydroxy functional group of SAL abolished its capability to yield 8-OH-dG in the presence of Cu(II). The dehydro analog of SAL, 1-methyl-6,7-dihydroxy-3,4-dihydroisoquinoline, produced significantly high levels of 8-OH-dG when incubated with calf thymus DNA, even in the absence of Cu(II), which appears to be attributable to the tautomer formation by this compound. In another experiment, SAL exerted cytotoxicity when treated to rat pheochromocytoma (PC12) cells. Based on these findings, it seems likely that SAL undergoes redox cycling in the presence of Cu(II) with concomitant production of ROS, particularly hydroxyl radical, which could contribute to DNA damaging and cytotoxic properties of this neurotoxin. PMID- 11390187 TI - Light-dependent generation of reactive oxygen species in cell culture media. AB - Cell culture media (RPMI 1640, Dulbecco's Minimal Essential Medium and yeast extract-peptone-glucose medium) were found to oxidize dichlorodihydrofluorescein diacetate and dihydrorhodamine 123, and to generate spin adduct of 5,5'-dimethyl 1-pyrroline N-oxide, which indicates formation of reactive oxygen species (ROS). The production of ROS was light dependent. The main component of the media responsible for the generation of ROS was riboflavin, but tryptophan, tyrosine, pyridoxine, and folic acid enhanced the effect of riboflavin. These observations point to exposure of cells to ROS under in vitro culture conditions. PMID- 11390188 TI - Activation of NFkappaB and MnSOD gene expression by free radical scavengers in human microvascular endothelial cells. AB - The effect of nonprotein thiol (NPT) free radical scavengers WR-1065 (SH) and WR 33278 (SS), the active thiol and disulfide metabolites of amifostine, N acetylcysteine (NAC; both L- and D- isomers), mesna, captopril, and dithiothreitol (DTT) on NFkappaB activation in human microvascular endothelial cells (HMEC) was investigated and contrasted to TNFalpha. The use of each of these NPTs at millimolar concentrations independent of oxidative damage-inducing agents resulted in a marked activation of NFkappaB, with the maximum effect observed between 30 min and 1 h after treatment. Only the SH and SS forms of amifostine, however, were effective in activating NFkappaB when administered at micromolar levels. Using a supershift assay, SH and SS equally affected the p50 p65 heterodimer, but not homodimers or heterodimers containing p52 or c-Rel subunits of NFkappaB. Neither catalase nor pyruvate when added to the culture medium to minimize hydrogen peroxide production had an effect on NFkappaB activation by SH. Thus, while oxidative damage is known to activate NFkappaB, the intracellular redox environment may also be affected by the addition of free radical scavenging agents such as NPT, and these in turn are capable of activating the redox sensitive transcription factor NFkappaB. There does not appear to be a significant role, if any, for the production of H(2)O(2) as an intermediate step in the activation of NFkappaB by either the SH or the SS form of amifostine. Rather, the underlying mechanism of action, especially for the SS form, may be related to the close structural and functional similarities of these agents to polyamines, which have been reported to be capable of activating NFkappaB. In contrast to TNFalpha, exposure of cells to either 40 microM or 4 mM of SH for 30 min did not induce intercellular adhesion molecule-1 (ICAM-1) gene expression, but did increase manganese superoxide dismutase (MnSOD) gene expression. MnSOD expression rose by 2-fold and remained elevated from 4 to 22 h following SH exposure. PMID- 11390189 TI - Rethinking cystic fibrosis pathology: the critical role of abnormal reduced glutathione (GSH) transport caused by CFTR mutation. AB - Though the cause of cystic fibrosis (CF) pathology is understood to be the mutation of the CFTR protein, it has been difficult to trace the exact mechanisms by which the pathology arises and progresses from the mutation. Recent research findings have noted that the CFTR channel is not only permeant to chloride anions, but other, larger organic anions, including reduced glutathione (GSH). This explains the longstanding finding of extracellular GSH deficit and dramatically reduced extracellular GSH:GSSG (glutathione disulfide) ratio found to be chronic and progressive in CF patients. Given the vital role of GSH as an antioxidant, a mucolytic, and a regulator of inflammation, immune response, and cell viability via its redox status in the human body, it is reasonable to hypothesize that this condition plays some role in the pathogenesis of CF. This hypothesis is advanced by comparing the literature on pathological phenomena associated with GSH deficiency to the literature documenting CF pathology, with striking similarities noted. Several puzzling hallmarks of CF pathology, including reduced exhaled NO, exaggerated inflammation with decreased immunocompetence, increased mucus viscoelasticity, and lack of appropriate apoptosis by infected epithelial cells, are better understood when abnormal GSH transport from epithelia (those without anion channels redundant to the CFTR at the apical surface) is added as an additional explanatory factor. Such epithelia should have normal levels of total glutathione (though perhaps with diminished GSH:GSSG ratio in the cytosol), but impaired GSH transport due to CFTR mutation should lead to progressive extracellular deficit of both total glutathione and GSH, and, hypothetically, GSH:GSSG ratio alteration or even total glutathione deficit in cells with redundant anion channels, such as leukocytes, lymphocytes, erythrocytes, and hepatocytes. Therapeutic implications, including alternative methods of GSH augmentation, are discussed. PMID- 11390190 TI - Spiral-shaped iatrogenic arterial dissection during superior mesenteric arteriography: a latent risk of diagnostic angiography using the power injector. AB - We report a rare case of iatrogenic arterial dissection of the superior mesenteric artery (SMA) during diagnostic angiography. A conventional superior mesenteric arteriogram obtained using an automated power injector revealed an arterial dissection 2s after the initiation of contrast-medium injection. This case indicates that although careful catheter manipulations during angiography are essential, certain unavoidable complications may occur. PMID- 11390191 TI - Spontaneous bilateral internal carotid artery dissection with hypoglossal nerve palsy. AB - Spontaneous dissection of the extracranial internal carotid artery (ICA) is increasingly being recognized as a common vascular disease. We report on a 52 year-old-male presented with bilateral extracranial internal artery dissection and twelfth nerve palsy and review the previous literature. PMID- 11390192 TI - A method of radio-frequency inhomogeneity correction for brain tissue segmentation in MRI. AB - An automatic method of correcting radio-frequency (RF) inhomogeneity in magnetic resonance images is presented. The method considers that image intensity variation due to radio-frequency inhomogeneity contains not only low frequency components, but also high frequency components. The variation is regarded as a multiplication of low frequency (capacity variation of coil) and the frequency of object (true image). The efficiency of the proposed method is illustrated with the aid of both phantom and physical images. The impact of the inhomogeneity correction on brain tissue segmentation is studied in detail. The results show significant improvement of the tissue segmentation after inhomogeneity correction. PMID- 11390193 TI - Herpes simplex encephalitis: diffusion MR imaging findings. AB - We studied five patients with herpes simplex encephalitis type 1 with diffusion weighted MR imaging, as this sequence provides information regarding tissue integrity. Diffusion-weighted imaging was obtained using the echo-planar sequence. True diffusion images (i.e. b=1000s/mm(2) images), and the ADC (apparent diffusion coefficient) values were studied. In addition, diffusion imaging of 12 normal cases, 46 patients with acute ischemia (cytotoxic edema), and 12 patients with vasogenic edema were studied for comparison. In patients with herpes encephalitis, two distinct types of diffusion imaging findings (on b=1000s/mm(2) images, and ADC maps) were noted: lesions similar to cytotoxic edema, and lesions similar to vasogenic edema. The patients with the former type of lesions had fulminating disease, and were in severe clinical condition. Those with the latter represented early cases, and they were in fairly good clinical condition with a good outcome by prompt therapy. Diffusion imaging appears to be a promising sequence to monitor the changes in the brain tissue in herpes encephalitis, and in other infections as well with respect to restriction of movement (cytotoxic edema) or relatively high-motion (vasogenic edema) of water molecules, providing data on the severity of the disease. PMID- 11390194 TI - Assessment of lung volumes in pulmonary emphysema using multidetector helical CT: comparison with pulmonary function tests. AB - The purpose of this study is to evaluate the usefulness of multidetector helical CT (MDCT) with three-dimensional (3D) postprocessing for assessing the lung volume at inspiration and expiration of the pulmonary emphysema and for comparing it with pulmonary function tests. Percentage lung volume at the threshold of 930, -900, -810, -790, and -770 at expiration showed good correlation with FEV1, FEV1/FVC, and DLCO/Va. Excellent correlation was observed between percentage lung volume at the threshold of -900 and FEV1/FVC. CT densitometry at expiration showed better correlation than that at inspiration with pulmonary function tests. MDCT with 3D technique is useful for assessing the severity of pulmonary emphysema. PMID- 11390195 TI - Growing-cube isosurface extraction algorithm for medical volume data. AB - In medical applications, three-dimensional volume data such as CT and MRI are gathered from medical-imaging devices. Marching cube (MC) algorithm is a common routine to extract isosurfaces from volume data. The MC algorithm generates the massive number of triangles to represent an isosurface. It is difficult to render this amount of triangles in real-time on general workstations. In this paper, we present a growing-cube algorithm to reduce the number of triangles generated by the MC algorithm. Growing-cube algorithm uses a surface tracker to avoid exhaustive searching isosurfaces cell-by-cell and, therefore, it saves computation time. During surface tracking, the growing-cube algorithm adaptively merges surfaces contained in the tracked cells to reduce the number of triangles. Surfaces are merged as long as the error is within user-specified error thresholds. Therefore, the proposed algorithm can generate a variable resolution of isosurfaces according to these error parameters. PMID- 11390196 TI - Proton MR spectroscopy of craniopharyngiomas. AB - To date, only a few cases of craniopharyngiomas have been studied by magnetic resonance (MR) spectroscopy. We report our spectroscopy experiences with five patients having surgically proven craniopharyngiomas. Proton MR spectroscopy images were obtained using the single-voxel mode with spin-echo point resolved spectroscopy. Very prominent peaks centered at 1-1.5ppm were noted in spectroscopic analysis, which probably corresponded to lipid/cholesterol peaks, correlating with the histological findings revealing high amounts of cholesterol in the cyst fluids. PMID- 11390197 TI - Congenital anomalies of the pancreaticobiliary tract: findings on MR cholangiopancreatography (MRCP) using half-Fourier-acquisition single-shot turbo spin-echo sequence (HASTE). AB - The clinical significance of magnetic resonance cholangiopancreatography (MRCP) using half-Fourier acquisition single-shot turbo spin-echo (HASTE) in the evaluation of congenital anomalies of pancreaticobiliary tract was demonstrated in 56 patients (man: 31, women: 25; age: 0-60 years old, mean 15 years old) with suspected congenital anomalies of pancreaticobiliary tract. The image quality and characteristics were assessed. MRCP using HASTE with a phased array coil can be used as a non-invasive method for the evaluation of congenital anomalies of the pancreaticobiliary tracts. PMID- 11390198 TI - A fast progressive method of maximum intensity projection. AB - Real-time processing and visualization of the 3D image data are the most important requirements for medical imaging. Among various 3D visualization methods, maximum intensity projection (MIP) is a useful tool to visualize 3D medical images. However, a large computation amount is a drawback of using the MIP image in clinical diagnosis. The processing time of the MIP depends on the number of voxels of the 3D data. In order to overcome the large amount of computation for the MIP, we have developed a progressive MIP method that can perform the MIP with low-resolution for fast processing, and use the low resolution MIP image to generate a full-resolution MIP image with a reduced computation time. In this paper, the progressive MIP method is implemented and its computation complexity is analyzed. PMID- 11390199 TI - Development of a tracking method for image analysis of time-lapse deformation of trabecular structure in a human bone. AB - A new method to track the deformation of the trabecular structure in a human bone, which determines the dynamic characteristics of bone, was developed using elastic matching. This is a kind of image analysis. By comparing two 3-D images of the same bone taken at different times, this method computes displacement of any points in the bone during the period. This method makes it possible to demonstrate the behavior of the trabecular structure to mechanical loads and to estimate changes in the strength of a bone. PMID- 11390200 TI - Cloning of the hamster androgen receptor gene. AB - Flank organs of male Golden Syrian hamsters contain sebaceous glands and hair follicles whose morphology and function are highly dependent on androgen, which makes these organs a useful model to study androgen action. In order to investigate molecular mechanisms of androgen action, we cloned a cDNA encoding the hamster androgen receptor (hamAR) by polymerase chain reaction (PCR) amplification of hamster testis cDNA. Nucleotide sequence analysis revealed that the cDNA has the capacity to encode a polypeptide of 900 amino acid. The deduced amino acid sequence was highly homologous to those of androgen receptors (AR) from other species. Western blot analysis of COS1 cells transfected with a vector expressing hamAR revealed that the recombinant ham AR was identical in size to that of endogeneous ham AR expressed in liver, sebaceous glands and testis. We further demonstrated that transfection of the hamAR expression vector into COS1 cells resulted in activation of a luciferase reporter gene containing multiple androgen responsive elements (ARE) in a testosterone-dependent manner. Availability of the recombinant hamAR clone along with the flank organ system should provide a more powerful tool than currently available to investigate androgen action at the molecular level. PMID- 11390201 TI - Novel mutations of ATP2A2 gene in Japanese patients of Darier's disease. AB - Darier's disease (DD) is a rare, dominantly inherited skin disorder with abnormal keratinization and acantholysis. Recently, mutations of ATP2A2 encoding the sarco/endoplasmic reticulum Ca(2+)-ATPase type 2 isoform (SERCA2) have been reported in Caucasian DD families. In the present study, we examined the ATP2A2 gene mutations of three sporadic (AS1,AS3,AS4) and one familial (AS2) Japanese DD patients. Sequence analysis revealed that the patients had novel mutations, one nonsense mutation (AS1 (C613X)) and three single base changes leading to amino acid substitutions (AS2 (L321F), AS3 (I274V), and AS4 (M719I)). These results demonstrate that distinct ATP2A2 gene mutations are present in Japanese DD patients. PMID- 11390202 TI - The correlation of MHC haplotype and development of Behcet's disease-like symptoms induced by herpes simplex virus in several inbred mouse strains. AB - It has been postulated that human leukocyte antigen (HLA)-B51 is associated with Behcet's disease. In previous study, we induced Behcet's disease-like symptoms in ICR mice inoculated with herpes simplex virus. In this study, several inbred mouse strains -- B10.BR (H-2k), B10.RIII (H-2r), C57BL/6 (H-2b), C3H/He (H-2k), Balb/c (H-2d) -- which had different types of major histocompatibility complex (MHC), were inoculated with herpes simplex virus type 1 (KOS strain) in order to demonstrate the role of histocompatibility antigen in the development of Behcet's disease-like symptoms. Behcet's disease-like symptoms developed in 40-50% of B10.BR, B10.RIII and C57BL/6 strains, but in only 2% of C3H/He and Balb/c. B10.BR and C3H/He strains had a common haplotype (H-2k) but the rate of manifestation was different. So, we conclude that MHC is not directly correlated with development of Behcet's disease-like symptoms in the animal model of herpes simplex virus induced Behcet's disease-like symptoms. PMID- 11390203 TI - Injection of human herpesvirus-8 in human skin engrafted on SCID mice induces Kaposi's sarcoma-like lesions. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus 8 (HHV-8) has been implicated in the development of Kaposi's sarcoma (KS) and several B-cell lymphoproliferative diseases. Serologic and molecular genetic association data has implicated HHV-8 as the causal agent of KS, but its role in the development of KS lesions is not understood. To examine the etiology of KS, HHV-8 was injected into normal human skin transplanted onto SCID mice. Injection of HHV-8 induced lesion formation that is morphologically and phenotypically consistent with KS, including the presence of angiogenesis and spindle-shaped cells latently infected with HHV-8. These findings suggest that HHV-8 is indeed the etiologic agent of KS, and that the virus plays an important role in initiation of this disease. PMID- 11390204 TI - Melatonin reduces X-ray irradiation-induced oxidative damages in cultured human skin fibroblasts. AB - Melatonin is a hormone with multiple functions in humans, produced by the pineal gland and stimulated by beta-adrenergic receptors. Melatonin has been shown to have radioprotection properties, but there has been little progress toward identifying the specific mechanisms of its action. To clarify the role of melatonin as a radioprotective compound, in response to X-ray irradiation, we investigated the effects of X-ray irradiation and melatonin on cytotoxicity, lipid peroxidation and alteration of the cell cycle in cultured skin fibroblast. An 8 Gy dose of X radiation resulted in cell death in 63% of irradiated cells, i.e. the cell viability was 37%. The damage was associated with lipid peroxidation of the cell membrane, as shown by the accumulation of malondialdehyde (MDA). By pre-incubation with melatonin (10(-5) M), a significant preventive effect was noted on the increase in the absolute number of surviving cells (up to 68% of cells were survived), and the levels of MDA were markedly decreased. These findings suggest a close correlation between an increase of lipid peroxidation and a rate of cell death. Morphological changes associated with apoptotic cell death were demonstrated by TEM. DNA flow-cytometry analysis revealed that X radiation increased pre-G1 apoptotic population by 7.6% compared to a very low level (1.3%) of non-irradiated cells. However, in the presence of melatonin, this apoptotic population decreased up to 4.5% at 10(-5) M. The p53 and p21 protein levels of skin fibroblasts increased 4 h after 8 Gy irradiation, but melatonin pretreatment did not change those levels. This study suggests that melatonin pretreatment inhibits radiation-induced apoptosis, and melatonin exerts its radioprotective effect by inhibition of lipid peroxidation and without any involvement of the p53/p21 pathway. PMID- 11390205 TI - Antigenic components of Malassezia species for immunoglobulin E antibodies in sera of patients with atopic dermatitis. AB - Antigenic components of Malassezia furfur, M. globosa, M. restricta, M. slooffiae, and M. sympodialis were studied for immunoglobulin E antibodies in sera of patients with atopic dermatitis (AD). Antigenic components were extracted from Malassezia cells by treatment with beta-mercaptoethanol, referred to as 2-ME extract. CBB staining and lectin blots using Con A, LCA, PHA-E4, PNA or RCA120 showed that the 2-ME extracts contained several species-dependent components that differed quantitatively and qualitatively among the Malassezia species at the protein level. In the Western blot with the 2-ME extracts, of 54 sera of the patients with AD (54 patients), the patients' IgE antibodies most frequently recognized components showing molecular weights of 43-46 kDa for M. slooffiae, 12 22 kDa for M. sympodialis, 35-40 kDa for M. restricta, 45-50 kDa for M. globosa, and of 67-72 kDa for M. furfur, respectively. In the correlative study, in which the total band intensities generated for each extract in Western blot were compared among the Malassezia species, the intensity for M. globosa was well correlated with that for M. sympodialis (r=0.756). In the Western blot inhibition test, the 2-ME extract of M. globosa partially inhibited the reaction of the antigenic components of other Malassezia species with the patient's IgE antibodies. These results indicated that Malassezia species contained both species-specific and common antigenic components at the IgE antibody level. PMID- 11390206 TI - Low-dose ultraviolet B radiation synergizes with TNF-alpha to induce apoptosis of keratinocytes. AB - High-dose ultraviolet B (UVB) irradiation is known to induce apoptosis of keratinocytes, but low-dose UVB dose not. In this paper we present evidence that low-dose UVB can induce TNF-alpha-dependent apoptosis of keratinocytes. In our study, 5 mJ/cm(2) doses of UVB were not sufficient by themselves to induce apoptosis of cultured human keratinocytes, but 20 mJ/cm(2) doses of UVB were. The combination of 5 mJ/cm(2) doses of UVB and exogenous TNF-alpha (15 ng/ml) induced significant apoptosis of keratinocytes, although exogenous TNF-alpha without UVB did not. This phenomenon was accompanied by enhanced clustering of tumor necrosis factor receptor 1 (TNFR1). TNF-alpha's promotion of the induction of apoptosis by low-dose UVB was seen until 30 min after irradiation but not at 1 h. We confirmed this finding using a skin organ culture system. UVB (20 mJ/cm(2)), which did not induce transformation of epidermal keratinocytes into sunburn cells, induced apoptosis when TNF-alpha was added to the culture medium. These results suggest that one of the possible mechanisms of inducing keratinocyte apoptosis by low dose UVB and TNF-alpha is that low-dose UVB augments ligand-binding-induced TNFR1 clustering, resulting in increased apoptotic cell death. PMID- 11390207 TI - Pre-exposure with low-dose UVA suppresses lesion development and enhances Th1 response in BALB/c mice infected with Leishmania (Leishmania) amazonensis. AB - This study was conducted to determine whether exposing mice to ultraviolet (UV) radiation would alter the pathogenesis of infection with Leishmania (Leishmania) amazonensis (L. amazonensis) which causes progressive cutaneous disease in susceptible mouse strains. BALB/c mice were irradiated with 10 and 30 J/cm(2) UVA on shaved skin of the back from Dermaray (M-DMR-100) for 4 consecutive days before infection with Leishmania promastigotes. The course of disease was recorded by measuring the size of lesions at various times after infection. Mice groups irradiated with UVA 10 and 30 J/cm(2) showed significantly suppressed lesion development compared with the non-irradiated mice. Light and electron microscopy revealed a few parasites at the site of inoculation in UVA-irradiated subjects. Sandwich enzyme-linked-immunosorbent-assay (ELISA) examination of sera showed dose dependently upregulated interferon-gamma (IFN-gamma), tumor necrosis factor-alpha (TNF-alpha) and interleukin (IL)-12, and downregulated interleukin (IL)-4 and interleukin (IL)-10 levels in UVA-irradiated as compared with the non irradiated mice. Positive signals for IFN-gamma mRNA in irradiated mice were obtained by RT-PCR, while non-irradiated mice showed negative results. None of the examined samples showed signal for IL-4 mRNA. The present study disclosed that exposure of mice to different low-doses of UVA irradiation prior to infection may interfere with immunity to L. amazonensis in the murine model. This indicates that the cell-mediated response switch from Th2 to Th1 pattern suppressed the cutaneous lesions of L. amazonensis. PMID- 11390208 TI - Biosensors for direct determination of organophosphate pesticides. AB - Direct, selective, rapid and simple determination of organophosphate pesticides has been achieved by integrating organophosphorus hydrolase with electrochemical and opitical transducers. Organophosphorus hydrolase catalyzes the hydrolysis of a wide range of organophosphate compounds, releasing an acid and an alcohol that can be detected directly. This article reviews development, characterization and applications of organophosphorus hydrolase-based potentiometric, amperometric and optical biosensors. PMID- 11390209 TI - Development of dual receptor biosensors: an analysis of FRET pairs. AB - The development of a dual receptor detection method for enhanced biosensor monitoring was investigated by analyzing potential fluorescent resonance energy transfer (FRET) pairs. The dual receptor scheme requires the integration of a chemical transducer system with two unique protein receptors that bind to a single biological agent. The two receptors are tagged with special molecular groups (donors and acceptors fluorophores) while the chemical transduction system relies on the well-known mechanisms of FRET. During the binding event, the two FRET labeled receptors dock at the binding sites on the surface of the biological agent. The resulting close proximity of the two fluorophores upon binding will initiate the energy transfer resulting in fluorescence. The paper focuses on the analysis and optimization of the chemical transduction system. A variety of FRET fluorophore pairs were tested in a spectrofluorimeter and promising FRET pairs were then tagged to the protein, avidin and its ligand, biotin. Due to their affinities, the FRET-tagged biomolecules combine in solution, resulting in a stable, fluorescent signal from the acceptor FRET dye with a simultaneous decrease in fluorescent signal from the donor FRET dye. The results indicate that the selected FRET pairs can be utilized in the development of dual receptor sensors. PMID- 11390210 TI - Acoustic and optical transduction of BuChE binding to procainamide modified surfaces. AB - A novel polymer, poly(procainamide), PPA, containing numerous binding sites for cholinesterases was synthesized as a recognition layer for butyryl cholinesterase (BuChE) interaction with the ligand procainamide, utilizing TSM and SPR sensors. The polymer was synthesized by the reaction of methacryloyl chloride and procainamide followed by radical polymerization. Sensor surfaces (Au or SiO(2)) were spin-coated by the polymer solution to form thin layers. Binding of BuChE was found to be sensitive to the drying procedure of the polymer layer. The binding of BuChE to the polymer coated sensors was monitored on-line by following the response of thickness shear mode (TSM) and surface plasmon resonance (SPR) sensors. Binding of BuChE to PPA-coated TSM sensors were shown to follow a Langmuir isotherm giving association constant 3.4x10(6) M(-1). PMID- 11390211 TI - Electrocatalytic reduction of lipoic acid and electroenzymatic reduction of NAD(P)(+) for integrated dehydrogenase biosensors. AB - Biosensors for the oxidized substrates of NAD(P)(+)-specific dehydrogenases demand the reductive recycling of the coenzymes. So far, suitable catalysts for the corresponding two-electron transfer are not available. In the present paper, this transport has been realized by a combined electrocatalytical and electroenzymatic process. Lipoic acid has been reduced on graphite electrodes functionalized with Fe(II)-phthalocyanine in 95% yield at-1200 mV in phosphate buffer pH 7.0. With the electrocatalytically reduced product, dihydrolipoic acid, lipoamide dehydrogenase could reduce NAD(+) in 20% yield and thioredoxin reductase NADP(+) in 18.4% yield. So far, the combined electrocatalytic/electroenzymatic system has not yet been realized, mainly because at the potential needed for the lipoic acid reduction, a parallel one electron reduction of NAD(P)(+) was observed, implying the dimerization of the coenzyme. PMID- 11390212 TI - Investigation of highly sensitive piezoelectric immunosensors for 2,4 dichlorophenoxyacetic acid. AB - The improved highly sensitive piezoelectric immunosensor has been developed and evaluated using a model interaction of antibody with the model hapten-herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). For immobilization of 2,4-D, the self assembled layers of cystamine, 4-aminothiophenol or 3,3'-dithio-bis(propionic acid N-hydroxysuccinimide ester) were formed on smooth and rough crystals coated with gold or silver electrodes. The immunochemical interactions performed well in all cases, the aminothiophenol on gold was chosen as the optimum with regard to regeneration of immunosensing surfaces. The kinetics of interaction of surface bound 2,4-D with free antibody provided significantly higher kinetic parameters (kinetic association rate constant) when using optically smooth crystals compared to common rough crystal. Therefore, the smooth crystal should be preferred for future kinetic studies. The competitive assay of the herbicide 2,4-D achieved a limit of detection of 10 ng/l using the monoclonal anti-2,4-D antibody F6C10. Finally, a direct assay format has been evaluated using a thicker layer of glutaraldehyde-crosslinked antibody on the sensing surface. The direct binding of a small herbicide molecule was followed in real time. The detected concentration of 2,4-D (5 microg/l) was low enough for future direct monitoring of this herbicide in water. PMID- 11390213 TI - A multicellular spheroid-based sensor for anti-cancer therapeutics. AB - The progress in cellular engineering offers novel approaches for anti-cancer therapies. To investigate the effectiveness of potential therapies efficient screening methods are required. We propose an impedance measurement system which enables the use of multicellular spheroid models in bioelectronic screening systems either for non destructive life-time diagnostic or anti-cancer therapies. A biohybrid sensor system is created comprising gene-manipulated T47D clone 11 breast carcinoma spheroids positioned hydrodynamically in a capillary system with electrodes. A novel approach employing an antisense-5'butyrylcholinesterase expression system is probed on reaggregated tumor cells under simulated microgravity, inhibiting the gene transcription and translation of the embryonic proliferation marker butyrylcholinesterase expressed in different tumor types. Alterations in the morphology of cell aggregates e.g. apoptosis or necrosis can be detected by impedance spectroscopy monitoring the electric behavior of membranes and extracellular space with a high resolution and reproducibility. The hydrodynamic positioning of 3D in vitro cell aggregates and the short time for the measurements represent an innovative method for a synchronized multicapillary screening system. The combination of the measuring system with a bioreactor enables cyclic life time recordings of impedance spectra for monitoring the cell aggregate properties for a long period. PMID- 11390214 TI - A long-term lifetime amperometric glucose sensor with a perfluorocarbon polymer coating. AB - We have developed an amperometric glucose sensor whose electrodes are coated with a four-layered membrane: 3-aminopropyltriethoxysilane (gamma-APTES), Nafion, glucose oxidase (GOX), and perfluorocarbon polymer (PFCP). Tests demonstrate the sensor's ability to accurately and successively determine glucose concentrations ranging from 2.8 to 167 mM, over a 66 day period with no increase in response time, while remaining imperviousness to the effects of interference species (2.8 mM ascorbic acid, 0.3 mM uric acid, 0.3 mM p-acetaminophen). Furthermore, tests on diabetic urine samples showed an excellent correlation coefficient of 0.985 (y=1.04x+4.73, n=30) between sensor results and those of Glucose-Dehydrogenase clinical laboratory analyses. PMID- 11390215 TI - A novel microphysiometer based on MLAPS for drugs screening. AB - This paper presents a novel microphysiometer for simultaneous measurements of several extracellular ions concentrations in living cells based on MLAPS (multi light addressable potentiometric sensor). In the microphysiometer, different sensitive membranes are illuminated in parallel with n light sources at different frequencies, the response amplitudes of each frequency component can be measured on-line by parallel processing algorithm. By the experiments, we can analyze the relations of the extracellular environmental H(+), Na(+), K(+), Ca(2+) under the effects of western medicines (dilantin, phenobarbital sodium, penicillin sodium) and Chinese drugs (scutellaria, medlar, hemlock parsley), and estimate the effects of several drugs. As the novel microphysiometer works under regular cell culture conditions, cells can be repeatedly simulated with drugs to complete dose response curve within a few hours. With the detection of a general parameter (extruded protons and ions), the system can be used to monitor the real-time process of the cells' metabolism, observe the functional responses of different kinds of membrane-bound receptors, evaluate the drugs. PMID- 11390216 TI - An optical biosensor employing tiron-immobilised polypyrrole films for estimating monophenolase activity in apple juice. AB - A method is described for the incorporation of tiron as a substrate for tyrosinase enzyme into a polypyrrole film deposited on indium titanium oxide (ITO) glass. The presence of tiron in the polypyrrole film is verified by cyclic voltammetry (CV). The enzyme activity using the polypyrrole-tiron film is confirmed by the catalytic conversion of immobilised substrate to quinones by the enzyme. The use of both potentiometric and optical methods for the detection of the catalytic activity of the polypyrrole-tiron film and their potential use for the determination of monophenolase activity of apple polyphenol oxidase is described. This is the first report of this kind whereby tiron has been immobilised in a polypyrrole matrix for the enzyme activity determination. PMID- 11390217 TI - Study of DNA hybridization on polypyrrole grafted with oligonucleotides by photocurrent spectroscopy. AB - Recognition of DNA sequences by biochemical sensor is generally performed by analysis after completion of hybridization. Using a technique able to directly translate the biological event into an electrical signal allows the in situ monitoring of the hybridization kinetics. In this aim, the photoelectrochemical behavior of one electroactive polymeric sensor based on a copolymer of polypyrrole and polypyrrole-oligonucleotide has been investigated in aqueous solution. This sensor has been studied as such (i) and in two other situations: (ii) when the copolymer is in presence of non-complementary oligonucleotides; and (iii) when the copolymer is in presence of complementary oligonucleotides. From the photocurrent spectra obtained at -0.6 V/SCE versus incident energy the allowed direct and indirect transitions for each polymer have been evidenced. The photocurrent evolution during hybridization and adsorption processes has been recorded in real time and the hybridization kinetics has revealed to be comparable with mass variations obtained by quartz crystal microbalance under the same experimental conditions. PMID- 11390218 TI - Monitoring of bacteria growth using a wireless, remote query resonant-circuit sensor: application to environmental sensing. AB - A new technique is presented for in-vivo remote query measurement of the complex permittivity spectra of a biological culture solution. A sensor comprised of a printed inductor-capacitor resonant-circuit is placed within the culture solution of interest, with the impedance spectrum of the sensor measured using a remotely located loop antenna; the complex permittivity spectra of the culture is calculated from the measured impedance spectrum. The remote query nature of the sensor platform enables, for example, the in-vivo real-time monitoring of bacteria or yeast growth from within sealed opaque containers. The wireless monitoring technique does not require a specific alignment between sensor and antenna. Results are presented for studies conducted on laboratory strains of Bacillus subtilis, Escherichia coli JM109, Pseudomonas putida and Saccharomyces cerevisiae. PMID- 11390219 TI - Porous silicon based potentiometric triglyceride biosensor. AB - A novel method for estimating triglycerides is reported. Porous silicon, prepared from p-type (100) crystalline silicon was thermally oxidized and used to immobilise lipase, an enzyme, which hydrolyses triglycerides resulting in the formation of fatty acids. This causes a change in the pH of the solution. Enzyme solution-oxidized porous silicon-crystalline silicon structure was used to detect changes in pH during the hydrolysis of tributyrin as a shift in the capacitance voltage (C-V) characteristics. Detailed calibration of the sensor is included. PMID- 11390220 TI - Recombinant Microdochium nivale carbohydrate oxidase and its application in an amperometric glucose sensor. AB - Biosensors containing recombinant carbohydrate oxidase from Microdochium nivale (rMnO) were developed by means of either chemically modified carbon paste or graphite electrode. 1-(N,N-dimethylamine)-4-(4-morpholine)benzene (AMB) and 1,1' dimethylferrocene (DMFc) have been used as mediators. The biosensors showed a linear calibration graph up to 18 mM of glucose when operated at 0.04-0.36 V versus a saturated calomel electrode. Almost no change was detected in the sensitivity of the biosensors at pH 7.2-8.1. The biosensors responded to other aldoses in the D-configuration, however, maximal sensitivity of the biosensor was towards D-glucose. The biosensor did not response to polyhydroxylic compounds such as D-mannitol, D-sorbitol and inositol. The advantages of the biosensors based on rMnO in comparison to Aspergillus niger glucose oxidase is a wider linear range, low sensitivity to oxygen and (in some cases) broad specificity. PMID- 11390221 TI - Bioelectric recognition assay (BERA). AB - A novel biosensory method has been developed for the determination of various chemical and biological molecules by assessing their electrophysiological interactions with a group of cells and cell components immobilized in a gel matrix that preserves their 'physiological' functions. The method was applied for the detection of: (i) hepatitis C virus in human blood samples; (ii) plant viruses; and (iii) a herbicide (glyphosate) in aqueous solutions. It was able to rapidly (assay time 3-5 min) and specifically detect the molecules in question at a concentration lower than 100 pg/ml, among other compounds f similar structure. The potential use of BERA biosensors for a rapid and cost-efficient molecule determination without prior knowledge of a specific receptor-molecule interaction is discussed. PMID- 11390223 TI - Genetic manipulation of spirochetes--light at the end of the tunnel. PMID- 11390225 TI - Now I know my CpGs. PMID- 11390226 TI - Anti-mosquito midgut antibodies block malaria transmission. PMID- 11390229 TI - Keeping bugs in shape. PMID- 11390231 TI - Exposure of the arsenal of another human pathogen. PMID- 11390233 TI - Ethiopia authorize drug imports. PMID- 11390234 TI - Potential AIDS vaccine? PMID- 11390240 TI - Is immune cell activation the missing link in the pathogenesis of post-diarrhoeal HUS? AB - Haemolytic uraemic syndrome (HUS), which is caused by Shiga toxin (Stx)-producing Escherichia coli, is the commonest cause of acute renal failure in childhood. It is widely believed that HUS develops following the release of Stx, an AB5 toxin that inhibits protein synthesis and has a direct toxic effect on the kidney endothelium. There remains, however, a mismatch between the current understanding of the pathogenesis of HUS and the evolution of the clinical signs of the disease. Our hypothesis is that Stx-mediated immune cell activation in the gut is the missing link in the pathogenesis of this condition, initiating the characteristic renal pathology of HUS either alone or in synergy with Stx. Validation of this hypothesis could lead to a targeted anti-inflammatory approach aimed at modulating immune cell function in HUS. PMID- 11390241 TI - Intracellular survival strategies of mutualistic and parasitic prokaryotes. AB - Endosymbiotic bacteria closely related to mammalian pathogens are widespread in invertebrates. Mutualistic and parasitic bacteria-host interactions on the various evolutionary levels apparently involve similar factors, indicating that relevant genetic information developed early in evolution. The detailed characterization of symbiotic interactions of bacteria with non-mammalian hosts should provide profound insights into the basic mechanisms of bacteria-host interactions and their evolution. PMID- 11390242 TI - Intracellular parasitism of macrophages by Cryptococcus neoformans. AB - Cryptococcus neoformans, an encapsulated fungal pathogen, causes meningoencephalitis in immunocompromised patients. Recent in vivo studies have demonstrated that C. neoformans is a facultative intracellular pathogen, as was previously suggested by in vitro studies. For survival in macrophages, C. neoformans utilizes a novel strategy for intracellular parasitism that includes the accumulation of intracellular polysaccharide in cytoplasmic vesicles. Confirmation of the fact that C. neoformans is a facultative intracellular pathogen could provide new insights into several poorly understood areas of cryptococcal pathogenesis, including mechanisms for latency and persistence and the lack of efficacy of humoral immunity. The finding that C. neoformans replicates inside macrophages in vitro in a manner similar to that observed in vivo provides an excellent system to dissect the molecular mechanisms responsible for this unique pathogenic strategy. PMID- 11390243 TI - Bartonella interactions with endothelial cells and erythrocytes. AB - Bartonella species are emerging human pathogens responsible for a wide range of clinical manifestations, including Carrion's disease, trench fever, cat-scratch disease, bacillary angiomatosis-peliosis, endocarditis and bacteraemia. During infection of their human or animal reservoir host(s), these arthropod-borne pathogens typically invade and persistently colonize mature erythrocytes. However, in both reservoir and incidentally infected hosts, endothelial cells are target cells for bartonellae. Endothelial interactions involve a unique mode of cellular invasion, the activation of a proinflammatory phenotype and the formation of vasoproliferative tumours. Based on the establishment of bacterial genetics and appropriate infection models, recent work has begun to elucidate the cell and molecular biology of these unusual pathogen-host cell interactions. PMID- 11390244 TI - Antibiotic and insecticide resistance modeling--is it time to start talking? AB - Mathematical models have played an important part in understanding both antibiotic and insecticide resistance. However, there has been little, if any, interdisciplinary work between these two areas of active research. One primary reason for this is that bacterial population genetics differ substantially from the population genetics of diploid organisms. This article examines these differences and their effect on resistance. It explores what efforts have gone into modeling resistance mathematically in both arenas, and offers suggestions on how the two groups could work together to gain a more comprehensive understanding of the resistance phenomenon PMID- 11390246 TI - Oxytocin and its receptor in the process of parturition. AB - OBJECTIVE: To review the potential roles of oxytocin (OT) and its receptor (OTR) in the regulation of parturition. METHODS: We reviewed the literature to describe the molecular aspects of the mechanisms of action and the regulation of OT and its receptor, particularly in relation to changes that occur at parturition. RESULTS: The literature provides strong support for a role for the OT/OTR system in the regulation of human parturition. Paracrine rather than endocrine mechanisms might be more important, and the regulation of the receptor might be more important than the ligand. The system is regulated by a wide variety of chemical and physical factors, including sex steroids, orphan receptors, uterine stretch, and many others. There also might be important interactions with the immune system. CONCLUSION: The OT/OTR system provides a potentially important therapeutic target for regulating the timing of human birth. PMID- 11390247 TI - Endotoxemia severely affects circulation during normoxia and asphyxia in immature fetal sheep. AB - OBJECTIVE: The purpose of the present study was to determine whether endotoxins (lipopolysaccharides, LPS) affect the fetal cardiovascular system in a way likely to cause brain damage. METHODS: Thirteen fetal sheep were chronically instrumented at a mean gestational age of 107 +/- 1 days. After control measurements of organ blood flow (microsphere method), blood gases, and acid base balance were obtained, seven of 13 fetuses received LPS (53 +/- 3 microg/kg fetal weight) intravenously. Sixty minutes later, asphyxia was induced by occlusion of the maternal aorta for 2 minutes. Measurements of organ blood flows were made at 60, -1, +2, +4, +30, and +60 minutes. RESULTS: Unlike in the control group, after LPS infusion there was a significant decrease in arterial oxygen saturation ( 46%; P <.001) and pH (P <.001). In LPS-treated fetuses the portion of combined ventricular output directed to the placenta decreased significantly (-76%; P <.001), whereas output to the fetal body (+60%; P <.001), heart (+167%; P <.05), and adrenals (+229%; P <.01) increased. Furthermore, during asphyxia circulatory centralization was impaired considerably in LPS-treated fetuses, and there was clear evidence of circulatory decentralization. This decentralization caused a severe decrease in cerebral oxygen delivery by 70%. Within 30 minutes after induction of asphyxia five of seven LPS-treated fetuses died, whereas all control fetuses recovered completely. CONCLUSIONS: Endotoxemia severely impaired fetal cardiovascular control during normoxia and asphyxia, resulting in a considerable decrease in cerebral oxygen delivery. These effects might have important effects in the development of fetal brain damage associated with intrauterine infection. PMID- 11390248 TI - Circulatory responses to acute asphyxia are not affected by the glutamate antagonist lubeluzole in fetal sheep near term. AB - OBJECTIVE: Asphyxia is one of the main causes of perinatal brain damage that can result in psychomotor deficits during later development. Recently lubeluzole, a new glutamate antagonist, was shown to improve clinical outcome considerably without any safety concerns in adults who had acute ischemic stroke. However, our preliminary experiments showed transient alterations in heart rate as well as arterial hypertension after intravenous application of this compound in fetal sheep. The aim of the present study was to examine in detail whether lubeluzole affects circulatory responses to acute asphyxia in fetal sheep near term. METHODS: Eleven fetal sheep were chronically instrumented at a mean gestational age of 133 +/- 2 days (term is at 147 days). The fetuses in the study group (n = 6) received three bolus injections of lubeluzole at 30-minute intervals (3 x 0.11 mg/kg estimated body weight), and five controls received solvent. Organ blood flows and physiologic variables were measured before, during, and after arrest of uterine blood flow for 2 minutes (ie, at 0, 1, 2, 3, 4, and 30 minutes). RESULTS: Before asphyxia, distribution of combined ventricular output and physiologic variables in fetuses from the control group were in the normal range for chronically prepared fetal sheep near term. During acute asphyxia there was a redistribution of cardiac output toward the central organs accompanied by pronounced bradycardia and progressive increase in arterial blood pressure. There were nearly no differences between groups in the time course of physiologic and cardiovascular variables measured before, during, and after acute intrauterine asphyxia. CONCLUSION: Lubeluzole did not affect circulatory responses to acute asphyxia in fetal sheep near term. PMID- 11390249 TI - Prenatal and postnatal ovine adrenal cell responses to prostaglandin E(2). AB - OBJECTIVE: To examine the secretory effect of prostaglandin E(2) (PGE(2)) and ACTH on the adrenal glands of prenatal and postnatal sheep. METHODS: Immunocytochemistry was used to examine the adrenal cortex and medulla for 17alpha-hydroxylase and tyrosine hydroxylase immunoreactivity. Microphysiometric technique was used to measure [H(+)] after exposure of whole dispersed prenatal and postnatal adrenal glands to PGE(2), ACTH, or both. RESULTS: Immunocytochemistry showed many cortical-type cells in all adrenal medullae and many medullary-type cells in fetal adrenal cortices. Maximum H(+) responsiveness to PGE(2) decreased with increasing age. The developmental age-related pattern of maximum percentage change in [H(+)] during ACTH exposure was similar to previous findings with cortisol production as the endpoint. ACTH stimulated H(+) production at 80 days' gestation and at all ages greater than 125 days' gestation (P <.05). The molar concentration of ligand required to elicit a response that was 50% of maximum response (EC(50)) for the ACTH response was lower in fetuses than in newborn lambs (<1 day and 3 days old), but there was no change in EC(50) for PGE(2) across the ages studied. Adrenal cell response to ACTH after prior ACTH and PGE(2) exposure was higher (P <.05) compared with ACTH after ACTH or ACTH alone at 110 days' gestation only and was lower in 3-day-old lambs. CONCLUSIONS: Based on the ACTH results, microphysiometry was a valid method for investigating dispersed adrenal cell physiology. Prostaglandin E(2) stimulated dispersed adrenal cells during the mid-gestation ACTH refractory period, but this effect decreased with increasing age. Prostaglandin E(2) sensitized adrenal cells to ACTH at 110 days' gestation but inhibited ACTH effects at postnatal day 3. PMID- 11390250 TI - Brain nitric oxide synthase expression is enhanced in the human cervix in labor. AB - OBJECTIVES: To determine whether brain nitric oxide synthase (bNOS) is expressed in human cervix, define its localization, and examine the possibility that it contributes to the nitric oxide (NO) pool and has a role in human cervical ripening. METHODS: Human cervical biopsy tissues were collected from four groups: (1) nonpregnant patients, (2) term nonlabor (cesarean delivery) patients, (3) term labor patients with cesarean deliveries, and (4) term labor patients (vaginal delivery). The mRNA expression was assessed using reverse-transcribed polymerase chain reaction and in situ hybridization. The protein expression was determined using Western blot and its localization was shown using immunostaining. RESULTS: Both bNOS mRNA and protein are present in human cervical tissue. They were localized to stroma cells and epithelial cells by in situ hybridization and immunostaining. bNOS protein expression in the labor group was significantly higher than in the nonlabor group (P <.05). CONCLUSION: bNOS is expressed in the human cervix and enhanced expression is observed in labor, suggesting its possible role in contributing to the NO pool in the human cervix, in cervical ripening, and in labor. PMID- 11390251 TI - Maternal plasma calcitonin gene-related peptide levels do not change during labor and are not influenced by delivery route. AB - OBJECTIVE: Calcitonin gene-related peptide (CGRP) circulates in maternal circulation throughout pregnancy, and specific receptors for CGRP (CGRPrs) are expressed by human myometrium. Because CGRP induces a dose-dependent relaxation of human myometrium, we examined a role for CGRP in modulation of myometrial smooth muscle contractility during pregnancy and labor. The aim of the present study was to evaluate the changes of maternal serum CGRP levels during parturition, according to the mode of delivery and in relation to cervical dilatation. METHODS: Circulating CGRP levels were measured in the following groups of healthy women: nonpregnant women, during the follicular phase of the menstrual cycle (n = 19); at term pregnancy (39-40 weeks; n = 24); after elective cesarean delivery (39-40 weeks; n = 20); and at spontaneous vaginal delivery (39 40 weeks; n = 16). In a subgroup of women, blood samples were collected longitudinally throughout labor at various cervical dilatations in the progress of labor (n = 8). RESULTS: Pregnant women at term not in labor had significantly higher CGRP levels than nonpregnant women (P =.021). No significant difference was found between women who delivered vaginally and those who had elective cesarean, and there were no correlations between CGRP plasma levels and cervical dilatation. CONCLUSIONS: Parturition is characterized by no significant changes in maternal serum CGRP levels, and no significant correlation exists between plasma CGRP levels and cervical dilatation during labor. PMID- 11390252 TI - Placental expression and serum levels of cytokeratin-18 are increased in women with preeclampsia. AB - OBJECTIVE: To evaluate placental expression and serum cytokeratin-18 in women with preeclampsia. METHODS: Serum cytokeratin-18 was evaluated in 44 women with preeclampsia and 44 healthy pregnant women using an immunoradiometric assay. Placental expression of cytokeratin-18 was investigated in specimens from 23 women with preeclampsia and 20 healthy pregnant women by immunohistochemistry. RESULTS: Median serum cytokeratin-18 in women with preeclampsia and healthy pregnant women was 106.7 and 76.0 U/L, respectively (P =.02). Among women with preeclampsia, serum cytokeratin-18 was significantly associated with severity of disease (P =.001) and showed a sensitivity (standard error) and specificity (standard error) of 85% (7%) and 65% (12%), respectively. In placental specimens, the cytoplasm of the syncytiotrophoblast stained positive for cytokeratin-18 with strong and widespread staining in 83% and 45% of placental specimens of women with preeclampsia and healthy pregnant women, respectively (P =.01). CONCLUSION: Elevated serum cytokeratin-18 values are associated with disease severity in women with preeclampsia. Our data provide additional evidence that the placenta might be the source of the elevated serum cytokeratin-18 values in women with preeclampsia. PMID- 11390253 TI - Prolonged inhibition of presynaptic catecholamine synthesis with alpha-methyl para-tyrosine attenuates the circadian rhythm of human TSH secretion. AB - OBJECTIVE: Originating from the pituitary gland, TSH secretion is regulated predominantly by thyroid-releasing hormone (TRH) neurons located in the hypothalamus. Norepinephrine and dopamine have important effects in modulation of TSH secretion. An inhibitor of catecholamine synthesis, alpha-methyl-para tyrosine (AMPT) has been used in several studies of the regulation of human TSH secretion. The short-term effects (<8 hours) of low doses of AMPT include stimulation of pituitary TSH secretion by selective lowering of brain dopamine levels. After prolonged administration of AMPT (>24 hours), theoretically both dopamine and norepinephrine levels are lowered significantly in the brain, although this has not been reported previously. METHODS: Nine subjects (five women and four men) received a total of five 1-g doses of AMPT or five 50-mg doses of promethazine (active placebo) over 28 hours in a randomized, double blind, placebo-controlled crossover design in which the active and control tests were separated by 4-6 weeks. Blood samples were obtained over 24 hours (18 time points) on day 2 of each condition. RESULTS: Changes in prolactin secretion and 6 hydroxymelatonin sulfate excretion indirectly showed the effects of AMPT on dopamine and norepinephrine. The typical circadian rhythm of TSH secretion was blunted by AMPT throughout the night; at ten time points, the difference between the two groups was statistically significant (P <.01). The long-term effects of repeated doses of AMPT were inhibition of TSH secretion and significant attenuation of the circadian rhythm of TSH. Additionally, AMPT induced low norepinephrine levels, which counteracted the stimulatory effect of low dopamine levels on TSH. CONCLUSION: Through its inhibitory effect on TRH, norepinephrine appeared to be involved in the regulation of TSH. PMID- 11390254 TI - Cyclin E mRNA overexpression in epithelial ovarian cancers: inverse correlation with p53 protein accumulation. AB - OBJECTIVE: We investigated the relationship between cyclin E mRNA overexpression and p53 protein accumulation in epithelial ovarian cancers. METHODS: mRNA was isolated and cDNA was prepared from 36 epithelial ovarian tumors (three adenomas, three low malignant potential tumors, and 30 carcinomas), and six normal ovaries. The cyclin E mRNA expression levels relative to an internal control, beta tubulin, were determined by semiquantitative polymerase chain reaction (PCR). Cyclin E and p53 protein expression in ovarian cancer tissues were examined by immunohistochemistry using the same series of samples. Fisher exact test of significance and an unpaired t test were used for statistical analysis. RESULTS: Considerable levels of cyclin E mRNA were detected in all normal ovaries and ovarian tumor samples examined by semiquantitative PCR amplification. mRNA levels of cyclin E were significantly higher in nine of 30 (30%) ovarian cancers compared with those in normal ovaries. The immunohistochemical expression of cyclin E protein was confirmed in the nuclei of tumor cells in 13 of 30 (43%) ovarian cancers. p53 protein accumulation was detected in 12 of 30 (40%) ovarian cancers examined. There was a significant inverse correlation between cyclin E mRNA overexpression and p53 protein accumulation (P <.01, Fisher exact test). CONCLUSIONS: Cyclin E mRNA overexpression frequently occurs in ovarian cancers without p53 protein accumulation. Cyclin E might have an important effect on the development of a limited number of ovarian cancers. PMID- 11390255 TI - Cell transplantation as a treatment for retinal disease. AB - It has been shown that photoreceptor degeneration can be limited in experimental animals by transplantation of fresh RPE to the subretinal space. There is also evidence that retinal cell transplants can be used to reconstruct retinal circuitry in dystrophic animals. Here we describe and review recent developments that highlight the necessary steps that should be taken prior to embarking on clinical trials in humans. PMID- 11390256 TI - Rod-cone interactions: developmental and clinical significance. AB - During the last decade, numerous research reports have considerably improved our knowledge about the physiopathology of retinal degenerations. Three non-mutually exclusive general areas dealing with therapeutic approaches have been proposed; gene therapy, pharmacology and retinal transplantations. The first approach involving correction of the initial mutation, will need a great deal of time and further development before becoming a therapeutic tool in human clinical practice. The observation that cone photoreceptors, even those seemingly unaffected by any described anomaly, die secondarily to rod disappearance related to mutations expressed specifically in the latter, led us to study the interactions between these two photoreceptor populations to search for possible causal links between rod degeneration and cone death. These in vivo and in vitro studies suggest that paracrine interactions between both cell types exist and that rods are necessary for continued cone survival. Since the role of cones in visual perception is essential, pending the identification of the factors mediating these interactions underway, rod replacement by transplantation and/or neuroprotection by trophic factors or alternative pharmacological means appear as promising approaches for limiting secondary cone loss in currently untreatable blinding conditions. PMID- 11390257 TI - Confronting complexity: the interlink of phototransduction and retinoid metabolism in the vertebrate retina. AB - Absorption of light by rhodopsin or cone pigments in photoreceptors triggers photoisomerization of their universal chromophore, 11-cis-retinal, to all-trans retinal. This photoreaction is the initial step in phototransduction that ultimately leads to the sensation of vision. Currently, a great deal of effort is directed toward elucidating mechanisms that return photoreceptors to the dark adapted state, and processes that restore rhodopsin and counterbalance the bleaching of rhodopsin. Most notably, enzymatic isomerization of all-trans retinal to 11-cis-retinal, called the visual cycle (or more properly the retinoid cycle), is required for regeneration of these visual pigments. Regeneration begins in rods and cones when all-trans-retinal is reduced to all-trans-retinol. The process continues in adjacent retinal pigment epithelial cells (RPE), where a complex set of reactions converts all-trans-retinol to 11-cis-retinal. Although remarkable progress has been made over the past decade in understanding the phototransduction cascade, our understanding of the retinoid cycle remains rudimentary. The aim of this review is to summarize recent developments in our current understanding of the retinoid cycle at the molecular level, and to examine the relevance of these reactions to phototransduction. PMID- 11390258 TI - Pattern electroretinography (PERG) and an integrated approach to visual pathway diagnosis. AB - The pattern electroretinogram (PERG) provides an objective measure of central retinal function, and has become an important element of the author's clinical visual electrophysiological practice. The PERG contains two main components, a positivity at approximately 50ms (P50) and a larger negativity at approximately 95ms (N95). The P50 component is affected by macular dysfunction with concomitant reduction in N95. The PERG therefore complements the Ganzfeld ERG in the assessment of patients with retinal disease. In contrast, the ganglion cell origins of the N95 component allow electrophysiological evaluation of ganglion cell function both in primary disease and in dysfunction secondary to optic nerve disease, where selective loss of N95 can be observed. Both macular dysfunction and optic nerve disease can give abnormalities in the visual evoked cortical potential (VEP), and the PERG thus facilitates more meaningful VEP interpretation. This review addresses the origins and recording of the PERG, and then draws on extensive clinical data from patients with genetically determined retinal and macular dystrophies, other retinal diseases and a variety of optic nerve disorders, to present an integrated approach to diagnosis. PMID- 11390260 TI - Comparison of plasma and peritoneal concentrations of various categories of MRI blood pool agents in a murine experimental pharmacokinetic model. AB - The aim of this study was to validate an experimental model designed to distinguish four categories of contrast agents, non specific agents (NDA, Gd DOTA) characterized by rapid and total extravasation; low diffusion agents (LDA, P760) characterized by delayed extravasation; and rapid (P792) and slow clearance (P717) blood pool agents (BPA) characterized by limited extravasation. Plasma and peritoneal gadolinium concentrations were simultaneously measured after intravenous injection of various contrast agents in mice. Products of each category were compared in this model.The plasma pharmacokinetic profiles were similar for Gd-DOTA and P760 (t1/2=13.3 and 13.8 min, respectively), whereas the half-lives were 22 and 1212 min for P792 and P717, respectively. The plasma clearance was inversely related to the size of the contrast agent. The intraperitoneal diffusion patterns of the various products were related to the molecular volume: C(max) per dose decreased progressively (78.7, 51.2, 44.2, 33.5 1/l) and t(max) increased (7, 15, 40, and 120 min) for Gd-DOTA, P760, P792 and P717, respectively. Nevertheless, the same quantities of Gd-DOTA and P760 (AUC ratio of 78.4 and 76.8, respectively) diffused into the peritoneum, whereas only 44.5% of P792 and 21.5% of P717 extravasated.The data obtained in this peritoneal permeability model with the various categories of contrast agents provide an estimation of the quantities of contrast agents diffusing into a permeable interstitium and may be used to predict the corresponding signal intensity, which can be measured locally. PMID- 11390261 TI - Magnetic resonance angiography at 0.3 T using MS-325. AB - Preliminary results on MS-325 versus ProHance enhanced magnetic resonance angiography (MRA) at low field strength in a rabbit model are reported. MS-325 enhanced images were acquired in vivo and compared with pre-contrast as well as conventional contrast-enhanced images. Visual image quality observations correlated with measurements of signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR). While published in vitro data show 7-fold greater relaxivity for MS 325 compared with conventional contrast agents, we observed an even greater effect here due, presumably, to better matching of the longer vascular lifetime with longer scan time in this study. In addition, overall vessel clarity improved significantly throughout all the phases of the experiment in MS-325-enhanced images when compared with conventional contrast-enhanced images. PMID- 11390262 TI - Deuterium NMR study of the MP-2269: albumin interaction--a step forward to the dynamics of non-covalent binding. AB - MP-2269, the Gd(III) complex of 4-pentylbicyclo[2.2.2]octane-1-carboxyl-di-L aspartyl-lysine-derived-DTPA, is a small Gd-agent that binds non-covalently to serum albumin in vivo to assume the enhanced relaxivities associated with macromolecular agents, (due in part to increased rotational correlation time, tau(R)). To further explore the fundamental parameters that govern the dynamics of water proton relaxation enhancement by this prototypical albumin-binding agent, the rotational correlation time (tau(R)) for the deuterated La(III) analog of MP-2269 has been independently measured in the presence and absence of 4% albumin using 2H-NMR approaches. The diamagnetic La(III) analog of MP-2269 was deuterated at the alpha-position of the carbonyl groups. 2H-NMR studies were conducted at 7.05T (46 MHz) and 310 degrees K on a Bruker NMR spectrometer. Spectral deconvolution permitted calculation of transverse relaxation rates, 1/T(2), from the NMR linewidths and subsequently, tau(R). The results yielded a tau(R) of the albumin bound complex of approximately 8 ns. This value is intermediate between those earlier estimated by 17O-NMR ( approximately 1 ns) and 1H-NMRD ( approximately 20-50 ns) and significantly shorter than that of albumin. The 2H-NMR study results also indicate that the exchange between free and albumin bound forms of the La(III) analog is slow (exchange lifetimes >1 ms). This slow exchange does not affect the water residence lifetimes (tau(M) 140-280 ns). PMID- 11390263 TI - Contrast agents in acute myocardial infarction. AB - The experimental design in examination of acute myocardial infarctions should be valid in terms of flow, perfusion and re-flow after intervention. The contrast agents concentration in experimental studies can be measured by microdialysis. We have assessed the usefulness of different extracellular and blood pool contrast agents for visualization of the area at risk in coronary artery occlusions. The double contrast technique, where Dy-DTPA-BMA was combined with Gd-DTPA-BMA yielded a superior infarct visualization. Blood pool agents for example NC100/150 injection is also promising in first path myocardial perfusion imaging. PMID- 11390264 TI - Pre-clinical results with Clariscan (NC100150 Injection); experience from different disease models. AB - A superparamagnetic nanoparticle (NC100150 Injection) was investigated in two different animal models; renal perfusion in pigs and tumour imaging in mice. In the pig model, qualitative first-pass perfusion maps following a bolus injection of NC100150 Injection enabled good visualisation of hypoperfused regions of the renal cortex following partial ligation of the renal artery. High temporal resolution was found to be essential to accurately capture the first passage of the contrast agent through the kidney due to the very rapid blood flow in normal renal cortex. In the tumour model (LS174T cells implanted in nude mice), NC100150 Injection was found to cause a gradual (over 60 min) signal increase on T1-w images in part of the tumours which was attributed to contrast agent leakage from the vascular space to the extravascular space in areas of increased capillary permeability. This observation is consistent with previous reports on the molecular cut-off size for vascular extraction for this tumour cell line. The specific enhancement of tumour tissue suggest potential utility of NC100150 Injection as an angiogenesis marker. PMID- 11390265 TI - Specific targeting of folate-dendrimer MRI contrast agents to the high affinity folate receptor expressed in ovarian tumor xenografts. AB - The need to develop target-specific MRI contrast agents to aid in disease characterization remains highly essential. In this study, we present a generation four polyamidoamine (PAMAM) folate-dendrimer that specifically targets the high affinity folate receptor (hFR) overexpressed on more than 80% of ovarian tumors. In vitro, mouse erythroleukemia cells expressing the hFR bind the radiolabeled folate-dendrimer chelate resulting in over 2700% increase in binding compared with untreated cells. The binding was inhibited by free folic acid to levels observed on folate-receptor-negative cells. In vivo, ovarian tumor xenografts resulted in a 33% contrast enhancement, following the folate-dendrimer chelate administration, that was significantly different compared with results obtained with a non-specific, extracellular fluid space agent, Gd-HP-DO3A. In addition, this contrast enhancement was absent in saline-treated animals, folate-receptor negative tumors, and was inhibited by free folic acid. Results suggest that a macromolecular, dendrimeric MRI agent with high molecular relaxivities (1646 mM( 1) s(-1)) can be used in specifically targeting the hFR on tumor cells and ovarian tumors. PMID- 11390266 TI - Mixed micelles containing lipophilic gadolinium complexes as MRA contrast agents. AB - Mixed micelles for MRA are multicomponent systems containing a phospholipid, a biocompatible non-ionic surfactant (e.g. Synperonic(R) F-108) and a lipophilic gadolinium complex. A variety of lipophilic gadolinium complexes were designed taking into account features such as: (i) nature of ligand (cyclic versus acyclic); (ii) lipophilic moiety; (iii) global charge of the complex; and (iv) nature of bond connecting the complex to the lipophilic moiety. All the lipophilic gadolinium complexes after formulation as mixed micelles show high relaxivities in water and in blood (rat). Mixed micelles containing gadolinium complexes bearing only one aliphatic chain cannot be used as MRA contrast agents because they have a high haemolytic effect. Furthermore, in rats they are quickly eliminated from the blood stream. These drawbacks are completely circumvented using gadolinium complexes bearing two aliphatic chains. Mixed micelles containing such complexes show high relaxivities, no haemolytic effect and long blood permanence. This makes them promising candidates as MRA contrast agents. However, elimination, which occurs exclusively through the liver, is not complete, even after 7 days. Complexes containing labile (e.g. ester) bonds between the lipophilic moieties and the chelate subunit are eliminated through both the liver and the kidneys. However, elimination is still not complete after 7 days. PMID- 11390268 TI - Pharmacokinetics of Gadomer-17, a new dendritic magnetic resonance contrast agent. AB - RATIONALE AND OBJECTIVES: Gadomer-17 is a new magnetic resonance (MR) contrast medium presently in clinical development. It is a dendritic gadolinium (Gd) chelate carrying 24 Gd ions. This study investigated the pharmacokinetic behavior of this contrast medium. METHODS: The pharmacokinetics of Gadomer-17 were investigated in different species (rat, rabbit, dog, monkey) for up to 7 days after intravenous (i.v.) injection of 25-100 micromol/kg body weight. In addition, elimination and biodistribution were evaluated after single i.v. injection of Gadomer-17 in rats. RESULTS: After i.v. injection Gadomer-17 distributes almost exclusively within the intravascular space without significant diffusion into the interstitial space. The volume of distribution (Vc) in the initial or alpha-phase ranged from 0.04 l/kg (rats, rabbits) to 0.06 l/kg (monkeys) and 0.07 l/kg (dogs), which reflects mainly the plasma volume. The blood/plasma concentration profile was found to be biphasic. The volume of distribution at a steady state is clearly smaller than that of other contrast media, which distribute to the extracellular space. After single i.v. injection in rats, the dendritic contrast medium was rapidly and completely eliminated from the body, mainly via glomerular filtration. No long-term accumulation or retention of the nonmetabolized agent was detectable in organs or tissues. CONCLUSIONS: Gadomer-17 is a promising new MR contrast medium that has an intravascular distribution and a rapid renal elimination. PMID- 11390267 TI - P792: a rapid clearance blood pool agent for magnetic resonance imaging: preliminary results. AB - An original MRI contrast agent, called P792, is described. P792 is a gadolinium macrocyclic compound based on a Gd-DOTA structure substituted by hydrophilic arms. The chemical structure of P792 has been optimized in order to provide (1) a high r(1) relaxivity in the clinical field for MRI: 29 mM(-1)xs(-1) at 60 MHz, (2) a high biocompatibility profile and (3) a high molecular volume: the apparent hydrodynamic volume of P792 is 125 times greater than that of Gd-DOTA. As a result of this high molecular volume, P792 presents an unusual pharmacokinetic profile, as it is a Rapid Clearance Blood Pool Agent (RCBPA) characterized by limited diffusion across the normal endothelium. The original pharmacokinetic properties of this RCBPA are expected to be well suited to MR coronary angiography, angiography, perfusion imaging (stress and rest), and permeability imaging (detection of ischemia and tumor grading). Further experimental imaging studies are ongoing to define the clinical value of this compound. PMID- 11390269 TI - Albumin-bound MRI contrast agents: the dilemma of the rotational correlation time. AB - Human serum albumin (HSA) binds numerous molecules, among which are suitably designed MRI contrast agents. The rotational tumbling of the protein is thus one of the parameters likely to affect the in vivo relaxivity of these agents. Literature unveils discrepancies about the value of the rotational correlation time (tau(R)) of HSA. In the present work, the tau(R) of this protein has been determined by studying the deuterium relaxation rate of small molecules known for their strong binding to HSA (warfarin and 4-hydroxycoumarin). Values of approx. 20-22 ns are obtained at 310 K in a 4% HSA solution and are in good agreement with the theoretical predictions. PMID- 11390270 TI - Java-based graphical user interface for the MRUI quantitation package. AB - This article describes the Java-based version of the magnetic resonance user interface (MRUI) quantitation package. This package allows MR spectroscopists to easily perform time-domain analysis of in vivo MR spectroscopy data. We show that the Java programming language is very well suited for developing highly interactive graphical software applications such as the MRUI software. We have also established that MR quantitation algorithms, programmed in other languages, can easily be embedded into the Java-based MRUI by using the Java native interface (JNI). This new graphical user interface (GUI) has been conceived for the processing of large data sets and uses prior knowledge data-bases to make interactive quantitation algorithms more userfriendly. PMID- 11390271 TI - Indirect evidence for the potential ability of magnetic resonance imaging to evaluate the myocardial iron content in patients with transfusional iron overload. AB - The purpose of this study was to evaluate the potential ability of magnetic resonance imaging (MRI) for evaluation of myocardial iron deposits. The applied MRI technique has earlier been validated for quantitative determination of the liver iron concentration. The method involves cardiac gating and may, therefore, also be used for simultaneous evaluation of myocardial iron. The tissue signal intensities were measured from spin echo images and the myocardium/muscle signal intensity ratio was determined. The SI ratio was converted to tissue iron concentration values based on a modified calibration curve from the liver model. The crucial steps of the method were optimized; i.e. recognition and selection of the myocardial slice for analysis and positioning of the regions of interest (ROIs) within the myocardium and the skeletal muscle. This made the myocardial MRI measurements sufficiently reproducible. We applied this method in 41 multiply transfused patients. Our data demonstrate significant positive linear relationships between different iron store parameters and the MRI-derived myocardial iron concentration, which was significantly related to the serum ferritin concentration (rho=0.62, P<0.0001) and to the MRI-determined liver iron concentration (rho=0.36, P=0.02). The myocardial MRI iron concentrations demonstrated also a significant positive correlation with the number of blood units given (rho=0.45, P=0.005) and the aminotransferase serum concentration (rho=0.54, P=0.0008). Our data represents indirect evidence for the ability of MRI techniques based on myocardium/muscle signal intensity ratio measurements to evaluate myocardial iron overload. PMID- 11390272 TI - Navigator motion correction of diffusion weighted 3D SSFP imaging. AB - Diffusion weighted (DW) 3D steady state MR (SSFP) head imaging technique using navigator echo's motion correction is presented. This new scheme enables acquisition of DW images even at regions where severe susceptibility is present. Another advantage is the moderate gradient performance requirements. DW imaging methods are sensitive to any kind of motion, thus, most of these methods might suffer from bulk motion artifacts. The common solution to avoid motion artifacts in a 2D DW SSFP acquisition is multi averaging. To avoid the time consuming multi averaging, the new scheme, described here, utilizes navigator echo's motion correction to remove respiratory bulk motion artifacts. At some brain regions, where the motion is governed by blood or CSF pulsation, the navigator motion correction fails. At these regions the correction is an interpolation of corrections from regions where the motion is particularly of the respiratory type. The combination of a 3D sequence with a navigator echo motion correction, enables acquisition of 10 DW slices within a time of 0:50-2:30 min. PMID- 11390273 TI - In vivo 1H MR spectra analysis by means of second derivative method. AB - Short echo time (TE) in vivo PRESS 1H MR spectra (2 T, TE=35 ms) of normal brain were fitted in the frequency domain using the second derivative method. In this approach, local maxima and hidden peaks are found as local minima of spectrum second derivative. The Lorentzian robust minimisation procedure (referred to as maximum likelihood or m-estimate fitting) using Levenburg-Marquardt non-linear fitting engine was applied. Spectral lines were approximated under the assumption of the mixed Lorentzian/Gaussian lineshapes. The same procedure was applied to 18 proton spectra. The number of peaks found within the range of 0.74/4.2 parts per million (ppm) was 52+/-3 and their positions were almost the same. The fitted lines were assigned on the basis of the J-pattern recalculated for the field strength of 2 T and by comparing the chemical shifts with the shifts in the single compound spectra. The ratios of main metabolites, such as NAA/Cr, Cho/Cr, Cho/NAA and mI/Cr, are in accord with those obtained earlier using the software supplied with the MR imager and the absolute concentrations of N-acetylaspartate (NAA), choline containing compounds (Cho), myoInositol (mI), glucose (Glc) and glutamate (Glu) obtained from the fit agree with those reported in literature, which confirms the usefulness of the second derivative method in routine analyses of 1H MR brain spectra. PMID- 11390274 TI - S100: a multigenic family of calcium-modulated proteins of the EF-hand type with intracellular and extracellular functional roles. AB - S100 is a multigenic family of non-ubiquitous Ca(2+)-modulated proteins of the EF hand type expressed in vertebrates exclusively and implicated in intracellular and extracellular regulatory activities. Within cells, most of S100 members exist in the form of antiparallelly packed homodimers (in some cases heterodimers), capable of functionally crossbridging two homologous or heterologous target proteins in a Ca(2+)-dependent (and, in some instances, Ca(2+)-independent) manner. S100 oligomers can also form, under the non-reducing conditions found in the extracellular space and/or within cells upon changes in the cell redox status. Within cells, S100 proteins have been implicated in the regulation of protein phosphorylation, some enzyme activities, the dynamics of cytoskeleton components, transcription factors, Ca(2+) homeostasis, and cell proliferation and differentiation. Certain S100 members are released into the extracellular space by an unknown mechanism. Extracellular S100 proteins stimulate neuronal survival and/or differentiation and astrocyte proliferation, cause neuronal death via apoptosis, and stimulate (in some cases) or inhibit (in other cases) the activity of inflammatory cells. A cell surface receptor, RAGE, has been identified on inflammatory cells and neurons for S100A12 and S100B, which transduces S100A12 and S100B effects. It is not known whether RAGE is a universal S100 receptor, S100 members interact with other cell surface receptors, or S100 protein interaction with other extracellular factors specifies the biological effects of a given S100 protein on a target cell. The variety of intracellular target proteins of S100 proteins and, in some cases, of a single S100 protein, and the cell specificity of expression of certain S100 members suggest that these proteins might have a role in the fine regulation of effector proteins and/or specific steps of signaling pathways/cellular functions. Future analyses should discriminate between functionally relevant S100 interactions with target proteins and in vitro observations devoid of physiological importance. PMID- 11390275 TI - Messenger RNA on the move: implications for cell polarity. AB - RNA sorting is an evolutionarily conserved mechanism for establishing asymmetries within a given cell concerning the macromolecular equipment of defined domains. mRNAs as well as non-coding transcripts are delivered to specific subcellular compartments in diverse organisms including developmental systems of Drosophila, Xenopus, ascidiens, zebrafish and echinoderms and in differentiated cells from yeast to mammals. The composition of the RNA localization machinery is complex. Both sequence- or structural motifs within RNA molecules to be transported (cis acting elements) and various proteins (trans-acting factors) contribute to the localization procedure. Most often these macromolecular ribonucleoprotein complexes exhibit a granular appearance, and granule localization depends on intact microfilaments or microtubules. When delivered to their ultimate destinations mRNAs are anchored to await translational activation at the appropriate timepoint. Beyond doubt, RNA localization plays a pivotal role in embryonic development, where mRNA mislocations cause severe body pattern defects. In terminally differentiated vertebrate cells RNA transport and local on-site translation presumably have an impact on various cellular functions such as cell motility, myelinization of nerve cell axons and nerve cell communications in the central and peripheral nervous system. PMID- 11390276 TI - On the promoting action of tamoxifen in a model of hepatocarcinogenesis induced by p-dimethylaminoazobenzene in CF1 mice. AB - BACKGROUND AND AIMS: Tamoxifen (TMX) has proven to be an effective palliative treatment for advanced breast cancer with low reported incidence of side effects. TMX has been demonstrated to be an initiator and/or a promoter in the rat model of hepatocarcinogenesis. To document the long-term effect of TMX in mice treated with p-dimethylaminoazobenzene (DAB), we have investigated the time response action of these drugs on different biochemical parameters. METHODS: A group of animals was placed on dietary DAB (0.5%, w/w) during a period of 28 weeks. Control animals received a standard laboratory diet. Two other groups of non treated and DAB-treated animals received TMX citrate (0.025%, w/w) in the diet since day 20. RESULTS: The activities of the enzymes involved in heme synthesis and degradation as evaluated in the DAB group was not further affected by TMX. DAB and/or TMX treatment significantly increased the content of total cytochrome P450 and also the activity of glutathione S-transferase indicating liver damage. In all treated groups oxidative stress and an adaptive response of the natural defense system (catalase and superoxide dismutase) were demonstrated. Histological and morphological studies revealed liver cell hyperplasia in DAB treated group; however, only in the DAB+TMX group solid, trabecular and acinar hepatocellular carcinoma was confirmed at the end of the experimental trial. CONCLUSION: We have demonstrated that TMX produced changes in hepatic enzyme activities which may be relevant for the metabolism and disposition of this and/or other drugs. Because liver tumors could be initiated and promoted by several agents which need to be activated, the possible hazard of TMX should be considered. This study reports that long-term treatment with TMX enhances hepatocarcinogenesis induced by DAB. The widespread use of TMX as an anticancer agent adds to the significance of this study. PMID- 11390277 TI - Purification and characterization of a proteinase inhibitor from white croaker skeletal muscle (Micropogon opercularis). AB - A trypsin proteinase inhibitor has been purified to homogeneity from the skeletal muscle of white croaker (Micropogon opercularis). Previously, we had described the occurrence in fish muscle of a serine protease (proteinase I) which showed a great capacity to degrade whole myofibrils in vitro and an endogenous inhibitor that prevented the action of the protease, both on natural and artificial substrates. In this paper, we report the purification and further biochemical characterization of the endogenous trypsin inhibitor. The purification was carried out by DEAE-Sephacel, Con A-Sepharose, Sephacryl S-300 and Mono Q. Throughout the purification procedure, trypsin inhibitory activity was assayed using azocasein as substrate. The molecular mass of the inhibitor was 65 kDa, as estimated by SDS-PAGE and gel filtration. The trypsin inhibitor is a glycoprotein, as deduced by the fact that it binds to Con A-Sepharose and stains with PAS and showed a wide range of pH stability (from 5 to 11). The thermal stability of the inhibitor considerably decreased at temperatures >60 degrees C. Assays of the inhibitor against various proteases indicated that it is highly specific for serine proteases, since it did not inhibit proteases belonging to any other groups. The inhibitor was able to inhibit the endogenous target enzyme (proteinase I) in a dose-dependent manner, with a 50% inhibition at a molar ratio close to 1. The present work contributes to improving our understanding of the physiological role of the proteinase I-inhibitor system in muscle protein breakdown, as well as its influence on post mortem proteolysis. PMID- 11390278 TI - Role of proline, glycerol, and heparin as protein folding aids during refolding of rabbit muscle creatine kinase. AB - Aggregation of 3 M guanidine hydrochloride denatured creatine kinase (ATP: creatine N-phosphotransferase, EC 2.7.3.2) occurs after dilution into the refolding solution. Proline, glycerol and heparin sodium act as folding aids which can effectively inhibit aggregation of creatine kinase during refolding. Proline at 1 M concentration, glycerol at 10% concentration and heparin at 25 mg/ml not only completely prevented creatine kinase aggregation but also enabled the creatine kinase to return to its native state as well as to recover most of its native activity. The reactivity after the aggregation was completely blocked by the presence of each folding aid reached 65-80% of the native activity. Results of turbidity, activity, intrinsic fluorescence and 1-anilinonaphthalene-8 sulfonate binding fluorescence measurements suggested that the effect of heparin differs from that of proline and glycerol in its artificial chaperone-like behavior. Heparin may bind with creatine kinase both in the native state and during the refolding course. The results showed that this heparin-creatine kinase complex favorably restored the creatine kinase reactivity. PMID- 11390279 TI - Evidence for a MARCKS-PKCalpha complex in skeletal muscle. AB - MARCKS (Myristoylated Alanine Rich C Kinase Substrate) is a protein known to cross-link actin filament and consequently, is very important in the stabilization of the cytoskeletal structure. In addition, it has been recently demonstrated that the phosphorylation rate of this protein changes during myogenesis and that this protein is implicated in fusion events. For a better understanding of the biological function of MARCKS during myogenesis, we have undertaken to identify and purify this protein from rabbit skeletal muscle. Three chromatographic steps including an affinity calmodulin-agarose column were performed. The existence of a complex between the two proteins was confirmed by non-denaturing gel electrophoresis and immunoprecipitation. Two complexes were isolated which present an apparent molecular weight of about 600 kDa. Such interactions suggest that MARCKS is either a very good PKCalpha substrate and/or a regulator of PKC activity. These results are supported by previous studies showing preferential interactions and co-localization of PKC isozyme and MARCKS at focal adhesion sites. This is the first time that MARCKS has been purified from skeletal muscle and our data are consistent with a major role of this actin- and calmodulin-binding protein in cytoskeletal rearrangement or other functions mediated by PKalpha. Our results provide evidence for a tight and specific association of MARCKS and PKCalpha (a major conventional PKC isozyme in skeletal muscle) as indicated by the co-purification of the two proteins. PMID- 11390280 TI - Regulation of alpha-smooth muscle actin gene expression in myofibroblast differentiation from rat lung fibroblasts. AB - Myofibroblasts express alpha-smooth muscle actin and have a phenotype intermediate between fibroblasts and smooth muscle cells. Their emergence can be induced by cytokines such as transforming growth factor beta; but the regulatory mechanism for induction of alpha-smooth muscle actin gene expression in myofibroblast differentiation has not been determined. To examine this mechanism at the level of the alpha-smooth muscle actin promoter, rat lung fibroblasts were transfected with varying lengths of the alpha-smooth muscle actin promoter linked to the chloramphenicol acetyl transferase reporter gene and treated with transforming growth factor beta1. The results show that the shortest inducible promoter was 150 base pairs long, suggesting the presence in this region of cis elements of potential importance in transforming growth factor beta1 induced myofibroblast differentiation. Transfection of "decoy" oligonucleotides corresponding to sequences for four suspected regulatory factors demonstrated that only the transforming growth factor beta control element is involved in the regulation of transforming growth factor beta1-induced alpha-smooth muscle actin expression in myofibroblast differentiation. Consistent with this conclusion is the finding that a mutation in the transforming growth factor beta control element caused a significant reduction in promoter activity. These observations taken together show that alpha-smooth muscle actin promoter regulation during myofibroblast differentiation is uniquely different from that in smooth muscle cells and other cell lines. Since myofibroblasts play a key role in wound contraction and synthesis of extracellular matrix, clarification of this differentiation mechanism should provide new insight into fibrogenesis and suggest future novel strategies for modulation of wound healing and controlling fibrosis. PMID- 11390281 TI - Semantic versus perceptual priming in fusiform cortex. PMID- 11390282 TI - The hippocampal complex and long-term memory revisited. AB - A recent report by Cipolotti et al. demontrates that the hippocampus and perhaps the parahippocampal area are essential for retrieval of remote episodic memory and important for remote semantic memory. This report, along with other recent findings, re-opens the debate about the role of these medial temporal lobe structures, indicating that their role extends much further than traditional theory had suggested. PMID- 11390283 TI - There's Waldo! PMID- 11390284 TI - Nothing to be afraid of? PMID- 11390285 TI - Mesmerizing studies. PMID- 11390286 TI - Measuring object recognition. PMID- 11390288 TI - Stimulating feedback. PMID- 11390287 TI - Cognitive science fiction. PMID- 11390289 TI - James or Jane? PMID- 11390291 TI - It's not what you look at, it's what you see. PMID- 11390290 TI - Animal concepts. PMID- 11390292 TI - Visual illusions and prefrontal cortex. PMID- 11390293 TI - Swimming with dolphins. PMID- 11390294 TI - Chunking mechanisms in human learning. AB - Pioneering work in the 1940s and 1950s suggested that the concept of 'chunking' might be important in many processes of perception, learning and cognition in humans and animals. We summarize here the major sources of evidence for chunking mechanisms, and consider how such mechanisms have been implemented in computational models of the learning process. We distinguish two forms of chunking: the first deliberate, under strategic control, and goal-oriented; the second automatic, continuous, and linked to perceptual processes. Recent work with discrimination-network computational models of long- and short-term memory (EPAM/CHREST) has produced a diverse range of applications of perceptual chunking. We focus on recent successes in verbal learning, expert memory, language acquisition and learning multiple representations, to illustrate the implementation and use of chunking mechanisms within contemporary models of human learning. PMID- 11390295 TI - Towards a distributed account of conceptual knowledge. AB - How is conceptual knowledge organized and represented? Are domains (such as living things) and categories (such as tools, fruit) represented explicitly or can domain and category structure emerge out of a distributed system? Taken at face value, evidence from brain-damaged patients and neuroimaging studies suggests that conceptual knowledge is explicitly structured in independent content-based stores. However, recent analyses of the fine-grained details of semantic impairments, combined with research using connectionist modelling, suggest a different picture - one in which concepts are represented as patterns of activation over multiple semantic properties within a unitary distributed system. Within this context, category-specific deficits emerge as a result of differences in the structure and content of concepts rather than from explicit divisions of conceptual knowledge in separate stores. PMID- 11390296 TI - Causes and consequences of imitation. AB - Recent behavioural and neuroscientific research concerning imitation has revealed evidence of experience-dependent imitation in chimpanzees and birds, wide ranging imitation deficits in autism, and unintentional imitation in adult humans. This review examines these findings and also evaluates evidence of neonatal imitation and intentional imitation in infancy, and evidence suggesting that the left inferior frontal gyrus is specialized for imitation. At the theoretical level, the empirical findings support the view that the perceptual-motor translation that is a unique and defining property of imitation depends primarily on direct links between sensory and motor representations established through correlated experience of observing movements and carrying them out. PMID- 11390297 TI - The shape of ears to come: dynamic coding of auditory space. AB - In order to pinpoint the location of a sound source, we make use of a variety of spatial cues that arise from the direction-dependent manner in which sounds interact with the head, torso and external ears. Accurate sound localization relies on the neural discrimination of tiny differences in the values of these cues and requires that the brain circuits involved be calibrated to the cues experienced by each individual. There is growing evidence that the capacity for recalibrating auditory localization continues well into adult life. Many details of how the brain represents auditory space and of how those representations are shaped by learning and experience remain elusive. However, it is becoming increasingly clear that the task of processing auditory spatial information is distributed over different regions of the brain, some working hierarchically, others independently and in parallel, and each apparently using different strategies for encoding sound source location. PMID- 11390298 TI - Decision making and neuropsychiatry. AB - Abnormal decision making is a central feature of neuropsychiatric disorders. Recent investigations of the neural substrates underlying decision making have involved qualitative assessment of the cognition of decision making in clinical lesion studies (in patients with frontal lobe dementia) and neuropsychiatric disorders such as mania, substance abuse and personality disorders. A neural network involving the orbitofrontal cortex, ventral striatum and modulatory ascending neurotransmitter systems has been identified as having a fundamental role in decision making and in the neural basis of neuropsychiatric diseases. This network accounts for the dissociations among decision-making deficits in different clinical populations. Ultimately, a more refined and sophisticated characterization of such deficits might guide the early diagnosis and cognitive and therapeutic rehabilitation of these patients. PMID- 11390299 TI - The 'feel' of seeing: an interview with J. Kevin O'Regan. PMID- 11390300 TI - Dynamics of water storage in mature subalpine Picea abies: temporal and spatial patterns of change in stem radius. AB - Internal water reserves in bark and foliage of trees contribute to transpiration (T) and play an essential role in optimizing water transport by buffering extreme peaks of water consumption. We examined patterns of stem shrinkage and their relationship to tree water dynamics. We measured fluctuations in root radius and stem radius at different stem heights, T of twigs at the top of the crown and sap flow velocities in stem sections of mature subalpine Norway spruce (Picea abies (L.) Karst.) trees over 2 years. The output of each sensor was coupled by physical functions to a mechanistic flow and storage model of tree water relations. The data verified the model-predicted lag in water storage depletion in response to the onset of transpiration and the lag increased with increasing distance from the crown periphery. Between the crown and stem base, the delay ranged from a few minutes to several hours, depending on microclimatic conditions and tree water status. Stem volume changes were proportional to the amount of water exchanged between the elastic tissues of the bark and the rigid xylem, indicating that the "peristaltic" wave of stem contraction along the flow path represented depletion of water stored in bark. On a daily basis, stems lost between 0.2 and 0.5% of their volume as a result of bark dehydration, corresponding to about 2 to 5 l of water. This water contributed directly to T. According to the model based on hydraulic principles, there are three main components underlying the dynamics of water storage depletion: flow resistance, storage capacities of needles and bark, and T of each tree section. The resistances and capacities were proportional to the response delay, whereas T in the lower parts of the tree was inversely proportional. The pattern of T within the crown depended on water intercepted by the branches. Because of these weather dependent factors, there was no time constant for the response delay along the flow path. Nevertheless, the upper crown and the root section tended to have longer response delays per meter of flow path than the stem. The diurnal course of stem radius fluctuations represents the sum of all external and internal conditions affecting tree water relations; stem radius fluctuations, therefore, provide a sensitive measure of tree water status. PMID- 11390301 TI - Responses of leaf respiration to temperature and leaf characteristics in three deciduous tree species vary with site water availability. AB - We measured responses of leaf respiration to temperature and leaf characteristics in three deciduous tree species (Quercus rubra L., Quercus prinus L. and Acer rubrum L.) at two sites differing in water availability within a single catchment in the Black Rock Forest, New York. The response of respiration to temperature differed significantly among the species. Acer rubrum displayed the smallest increase in respiration with increasing temperature. Corresponding Q(10) values ranged from 1.5 in A. rubrum to 2.1 in Q. prinus. Dark respiration at ambient air temperatures, expressed on a leaf area basis (Rarea), did not differ significantly between species, but it was significantly lower (P < 0.01) in trees at the wetter (lower) site than at the drier (upper) site (Q. rubra: 0.8 versus 1.1 micromol m(-2) s(-1); Q. prinus: 0.95 versus 1.2 micromol m(-2) s(-1)). In contrast, when expressed on a leaf mass basis (R(mass)), respiration rates were significantly higher (P < 0.01) in A. rubrum (12.5-14.6 micromol CO(2) kg(-1) s( 1)) than in Q. rubra (8.6-9.9 micromol CO(2) kg(-1) s(-1)) and Q. prinus (9.2 10.6 micromol CO(2) kg(-1) s(-1)) at both the lower and upper sites. Respiration on a nitrogen basis (R(N)) displayed a similar response to R(mass). The consistency in R(mass) and R(N) between sites indicates a strong coupling between factors influencing respiration and those affecting leaf characteristics. Finally, the relationships between dark respiration and A(max) differed between sites. Trees at the upper site had higher rates of leaf respiration and lower A(max) than trees at the lower site. This shift in the balance of carbon gain and loss clearly limits carbon acquisition by trees at sites of low water availability, particularly in the case of A. rubrum. PMID- 11390302 TI - Shoot water relations of mature black spruce families displaying a genotype x environment interaction in growth rate. III. Diurnal patterns as influenced by vapor pressure deficit and internal water status. AB - Pressure-volume curves were constructed and shoot water potentials measured for +20-year-old black spruce (Picea mariana (Mill.) BSP) trees from four full-sib families growing on a moist site and a dry site at the Petawawa Research Forest, Ontario, to determine whether differences in diurnal water relations traits were related to productivity. To assess the basis for the observed diurnal patterns, we analyzed effects of environmental and internal water stress variables on diurnal water relations traits. Among the water relations traits examined, turgor pressure was the most sensitive, responding to site, family and environmental variables and displaying the strongest diurnal responses to varying soil water availability and atmospheric vapor pressure deficit (VPD). Overall, there was an 84% drop in turgor pressure with increasing VPD: turgor pressure fell 46% in response to the first 0.75 kPa increase in VPD, and 9.7% in response to a second 0.75 kPa increase in VPD. The families differed in water relations responses to moderate water stress, but not in responses to minor or more extreme water stresses. Thus, at a VPD of 0.5 kPa, there was an estimated 83% greater family difference in turgor pressure on the dry site compared with the moist site. Soil and atmospheric water stress appeared to exert effects in tandem to elicit these responses (r(2) = 0.728). A comparison of the mechanisms of response to water deficit indicated that osmotic adjustment was more important than change in cell wall elasticity. We used a conceptual water relations model to illustrate the differences between tolerant and intolerant families in their mechanisms of water stress response. We conclude that, because genetic responses to site factors are dynamic, the integrated response over time contributes to the observed genetic x environmental interaction in growth. PMID- 11390303 TI - An improved heat pulse method to measure low and reverse rates of sap flow in woody plants. AB - The compensation heat pulse method (CHPM) is of limited value for measuring low rates of sap flow in woody plants. Recent application of the CHPM to woody roots has further illustrated some of the constraints of this technique. Here we present an improved heat pulse method, termed the heat ratio method (HRM), to measure low and reverse rates of sap flow in woody plants. The HRM has several important advantages over the CHPM, including improved measurement range and resolution, protocols to correct for physical and thermal errors in sensor deployment, and a simple linear function to describe wound effects. We describe the theory and methodological protocols of the HRM, provide wound correction coefficients, and validate the reliability and accuracy of the technique against gravimetric measurements of transpiration. PMID- 11390304 TI - Growth, leaf morphology, water use and tissue water relations of Eucalyptus globulus clones in response to water deficit. AB - Changes in leaf size, specific leaf area (SLA), transpiration and tissue water relations were studied in leaves of rooted cuttings of selected clones of Eucalyptus globulus Labill. subjected to well-watered or drought conditions in a greenhouse. Significant differences between clones were found in leaf expansion and transpiration. There was a significant clone x treatment interaction on SLA. Water stress significantly reduced osmotic potential at the turgor loss point (Pi0) and at full turgor (Pi100), and significantly increased relative water content at the turgor loss point and maximum bulk elastic modulus. Differences in tissue water relations between clones were significant only in the mild drought treatment. Among clones in the drought treatments, the highest leaf expansion and the highest increase in transpiration during the experiment were measured in those clones that showed an early and large decrease in Pi0 and Pi100. PMID- 11390305 TI - Atmospheric carbon dioxide, irrigation, and fertilization effects on phenolic and nitrogen concentrations in loblolly pine (Pinus taeda) needles. AB - Concentrations of total soluble phenolics, catechin, proanthocyanidins (PA), lignin and nitrogen (N) were measured in loblolly pine (Pinus taeda L.) needles exposed to either ambient CO(2) concentration ([CO(2)]), ambient plus 175 or ambient plus 350 micromol CO(2) mol(-1) in branch chambers for 2 years. The CO(2) treatments were superimposed on a 2 x 2 factorial combination of irrigation and fertilization treatments. In addition, we compared the effects of branch chambers and open-top chambers on needle chemistry. Proanthocyanidin and N concentrations were measured in needles from branch chambers and from trees in open-top chambers exposed concurrently for two years to either ambient [CO(2)] or ambient plus 200 micromol CO(2) mol(-1) in combination with a fertilization treatment. In the branch chambers, concentrations of total soluble phenolics in needles generally increased with needle age. Concentrations of total soluble phenolics, catechin and PA in needle extracts increased about 11% in response to the elevated [CO(2)] treatments. There were no significant treatment effects on foliar lignin concentrations. Nitrogen concentrations were about 10% lower in needles from the elevated [CO(2)] treatments than in needles from the ambient [CO(2)] treatments. Soluble phenolic and PA concentrations were higher in the control and irrigated soil treatments in about half of the comparisons; otherwise, differences were not statistically significant. Needle N concentrations increased 23% in response to fertilization. Treatment effects on PA and N concentrations were similar between branch and open-top chambers, although in this part of the study N concentrations were not significantly affected by the CO(2) treatments in either the branch or open-top chambers. We conclude that elevated [CO(2)] and low N availability affected foliar chemical composition, which could in turn affect plant-pathogen interactions, decomposition rates and mineral nutrient cycling. PMID- 11390306 TI - Differences in nitrogen economy of temperate trees. AB - Twenty-four temperate tree species were classified into three groups based on cluster analysis of relative growth rate, nitrogen concentration, nitrogen production efficiency, nitrogen-distribution ratio and nitrogen-use efficiency as follows: Group I (Asteridae and Rosidae), Group II (Dilleniidae and Hamamelidae) and Group III (Coniferopsidae). Relative growth rate (RGR) was high in Group II, moderate in Group I and low in Group III. The regression coefficient for the relationship between RGR and leaf nitrogen concentration was higher in Group II than in Group I, and no relationship was observed in Group III. Parameter analysis of RGR indicated that RGR per unit leaf nitrogen was important for all three groups, but that the allocation of nitrogen to leaves was particularly important in Groups I and II. The ratio of dark respiratory rate (R) to net photosynthetic rate (A) was higher in Group I than in Group II. Neither A nor R was measured in the Group III species. A linear relationship was observed between leaf nitrogen concentration and A in Group II, but this relationship was not evident in Group I. PMID- 11390307 TI - Characterization of proteinase activity in stratified Douglas-fir seeds. AB - We investigated the effect of stratification on the proteinase activity involved in mobilization of the major soluble approximately 45 kDa storage protein during germination of Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) seeds. Complete hydrolysis of the approximately 45 kDa protein was observed approximately 7 days after exposure of stratified seeds to germination conditions. Coincident with the onset of mobilization, proteinase activity was detected primarily in microsomal extracts from stratified seeds. Microsomal-associated proteinase activity was most active at pH 8.0 and had a molecular mass > 175 kDa as determined by gelatin SDS-PAGE gels. In vitro digestion of soluble protein extracts indicated that, following stratification, there was a significant increase in proteinase activity and hydrolysis of the approximately 45 kDa storage protein. Whether this increase was a result of activation of preexisting proteinase(s) or de novo synthesis remains unknown. In vitro digestion of soluble protein extracts in the presence of various proteinase inhibitors showed that digestion of the approximately 45 kDa storage protein is mediated primarily by a metalloproteinase and to a lesser degree by a serine proteinase. The accumulation of approximately 25 kDa protein products following in vitro digestion suggests that mobilization of the approximately 45 kDa soluble storage protein is mediated by a multi-step process involving the action of different classes of proteinases. PMID- 11390308 TI - Risks of stroke and current indications for cerebral revascularization in patients with carotid occlusion. AB - Preventing further stroke in patients with complete carotid artery occlusion remains a difficult challenge because there is no therapy proven effective for this prevention. These patients comprise approximately 15% of patients with carotid artery territory transient ischemic attacks or infarction. Patients with symptomatic carotid artery occlusion have an overall risk of subsequent stroke of 7% per year and a risk of stroke ipsilateral to the occluded carotid artery of 5.9% per year. The presence of severe hemodynamic failure demonstrated by increased oxygen extraction fraction (OEF) of the brain, in a cerebral hemisphere distal to a symptomatic occluded carotid artery, is an independent predictor of subsequent ischemic stroke with a risk comparable to that seen in medically treated patients with symptomatic severe carotid artery stenosis. PMID- 11390309 TI - Cerebral revascularization for cerebral ischemia and stroke prevention in patients with atherosclerotic occlusive disease: defining the population that benefits. AB - The decision to perform a revascularization procedure on a patient who may or may not benefit from it must be based on an understanding of the impact of spontaneous or therapeutic vascular occlusion on the physiologic, by which the cerebral circulation compensates for vascular occlusions. This impact is patient specific and requires examination of a number of possible physiologic parameters. PMID- 11390310 TI - Computerized hemodynamic evaluation of the cerebral circulation for bypass. AB - This article describes details of the application of computer modeling of cerebral blood circulation. A brief review of the different computer modeling techniques and the current models used today for predicting surgical options for bypass surgery are presented. The use of phase contrast MR for estimating intracranial flow rates makes these models even more accurate. Two case studies are presented with computer simulation results with verification of predicted outcome both clinically and from actual flow of measurements. PMID- 11390311 TI - Techniques for cerebral bypass: practical laboratory for microvascular anastomosis. AB - The techniques used for microvascular anastomosis are best mastered in the laboratory setting. A variety of techniques for end-to-end and end-to-side anastomoses are described and illustrated. An understanding of these different modalities allows the surgeon to choose the appropriate technique for each clinical setting. PMID- 11390312 TI - Posterior cerebral circulation revascularization. AB - Posterior circulation revascularization has evolved as a method to treat selected patients with vertebrobasilar ischemia who have inaccessible atherosclerotic occlusive disease and who have failed maximal medical therapy. In addition, complex unclippable aneurysms of the posterior circulation are another indication for revascularization of the vertebrobasilar territory. Careful preoperative evaluation and meticulous attention to detail intraoperatively yield good patient outcomes with minimal morbidity and mortality. This article reviews the vascular anatomy of the posterior circulation and the indications, preoperative evaluation, operative techniques, clinical outcomes, and alternative treatments for patients requiring posterior circulation revascularization procedures. PMID- 11390313 TI - The role of cerebral revascularization in patients with intracranial aneurysms. AB - Cerebral revascularization offers an important adjunct to parent-vessel ligation in the treatment of large and otherwise inaccessible intracranial aneurysms. Good or excellent outcomes can be expected in approximately 80% of patients. Poor outcomes and ischemic complications were highest in posterior circulation lesions. Cranial neuropathies from mass effect associated with giant aneurysms of the cavernous and intradural internal carotid artery will improve and be cured in the majority of patients treated with universal revascularization approach. In combination with open parent vessel ligation or endovascular occlusion, durable protection from subarachnoid hemorrhage can be achieved. PMID- 11390314 TI - Cerebral revascularization in cranial base tumors. AB - Certain cranial base tumors may involve intracranial arteries by encasement or invasion. In such patients, resection of the tumor along with the involved artery is an option for treatment. Techniques and results of vascular bypasses for such lesions are discussed in this article. PMID- 11390315 TI - Cerebral revascularization for moyamoya disease in children. AB - Ninety-four patients with moyamoya disease (56 in the pediatric age group and 38 adults) were treated by revascularization surgery in the past 21 years (1979- 2000). Combined surgery of the superficial temporal artery-middle cerebral artery (STA-MCA) anastomosis and indirect revascularization of encaphalo-duro-arterio myo-synangiosis (EDAMS) was performed. Ischemic attacks disappeared in most patients within 1 year (mean) after surgery in pediatric cases. No morbidity or mortality was experienced in the pediatric group. Some children with cerebral infarction before the surgery, however, had mild mental retardation even after the surgery. Early diagnosis and proper prophylactic surgical treatment is indispensable for pediatric patients with moyamoya. MR angiography is an important diagnostic modality for the screening and longitudinal follow-up of moyamoya disease. PMID- 11390316 TI - Cerebral revascularization for moyamoya disease in adults. AB - Moyamoya disease is a rare entity that results in progressive occlusion of the arteries of the circle of Willis. In adults, this most commonly leads to intraventricular hemorrhage and less frequently to symptoms of ischemia. Without treatment, there is progressive deterioration of neurologic function and re hemorrhage. Direct superficial temporal artery to middle cerebral artery bypass is considered the treatment of choice, although it's efficacy, particularly for hemorrhagic disease, remains uncertain. PMID- 11390317 TI - Cerebral revascularization in trauma and carotid occlusion. AB - Cerebrovascular disruption frequently results from head and neck trauma. Injury to the extra- and intracranial carotid artery is uncommon but is associated with a high rate of death and permanent neurologic deficit. In this article, injuries to the carotid artery are reviewed with emphasis on the mechanisms, clinical manifestations, radiologic evaluation, and management of these traumatic lesions. PMID- 11390318 TI - Preview of a new trial of extracranial-to-intracranial arterial anastomosis: the carotid occlusion surgery study. AB - In 1985, the International Study of Extracranial-to-Intracranial Arterial Anastomosis demonstrated no benefit from extracranial-to-intracranial arterial bypass operations in treatment of patients with extensive cerebrovascular disease including those with occlusions of the internal carotid artery. Interest in the potential use of extracranial-to-intracranial arterial bypass operations, however, has been rekindled by evidence that some patients with occlusion of the internal carotid artery have a poor collateral circulation and a high risk for recurrent ischemic events. Other patients with adequate perfusion after occlusion have a low likelihood for recurrent stroke. Restricting surgical treatment to only those patients judged to have a high risk for recurrent stroke might improve the usefulness of the bypass operation. A new clinical trial is proposed, testing the potential usefulness of extracranial-to-intracranial arterial bypass operations for treatment of carefully selected patients with occlusion of the internal carotid artery. Several issues that are being addressed in this new trial are described in this article. PMID- 11390319 TI - Health effects of dioxin exposure: a 20-year mortality study. AB - Follow-up of the population exposed to dioxin after the 1976 accident in Seveso, Italy, was extended to 1996. During the entire observation period, all-cause and all-cancer mortality did not increase. Fifteen years after the accident, mortality among men in high-exposure zones A (804 inhabitants) and B (5,941 inhabitants) increased from all cancers (rate ratio (RR) = 1.3, 95% confidence interval (CI): 1.0, 1.7), rectal cancer (RR = 2.4, 95% CI: 1.2, 4.6), and lung cancer (RR = 1.3, 95% CI: 1.0, 1.7), with no latency-related pattern for rectal or lung cancer. An excess of lymphohemopoietic neoplasms was found in both genders (RR = 1.7, 95% CI: 1.2, 2.5). Hodgkin's disease risk was elevated in the first 10-year observation period (RR = 4.9, 95% CI: 1.5, 16.4), whereas the highest increase for non-Hodgkin's lymphoma (RR = 2.8, 95% CI: 1.1, 7.0) and myeloid leukemia (RR = 3.8, 95% CI: 1.2, 12.5) occurred after 15 years. No soft tissue sarcoma cases were found in these zones (0.8 expected). An overall increase in diabetes was reported, notably among women (RR = 2.4, 95% CI: 1.2, 4.6). Chronic circulatory and respiratory diseases were moderately increased, suggesting a link with accident-related stressors and chemical exposure. Results support evaluation of dioxin as carcinogenic to humans and corroborate the hypotheses of its association with other health outcomes, including cardiovascular- and endocrine-related effects. PMID- 11390320 TI - Invited commentary: how do the Seveso findings affect conclusions concerning TCDD as a human carcinogen? PMID- 11390322 TI - Assessment of deaths attributable to air pollution: should we use risk estimates based on time series or on cohort studies? AB - Epidemiologic studies are crucial to the estimation of numbers of deaths attributable to air pollution. In this paper, the authors present a framework for distinguishing estimates of attributable cases based on time-series studies from those based on cohort studies, the latter being 5-10 times larger. The authors distinguish four categories of death associated with air pollution: A) air pollution increases both the risk of underlying diseases leading to frailty and the short term risk of death among the frail; B) air pollution increases the risk of chronic diseases leading to frailty but is unrelated to timing of death; C) air pollution is unrelated to risk of chronic diseases but short term exposure increases mortality among persons who are frail; and D) neither underlying chronic disease nor the event of death is related to air pollution exposure. Time series approaches capture deaths from categories A and C, whereas cohort studies assess cases from categories A, B, and C. In addition, years of life lost can only be derived from cohort studies, where time to death is the outcome, while in time-series studies, death is a once-only event (no dimension in time). The authors conclude that time-series analyses underestimate cases of death attributable to air pollution and that assessment of the impact of air pollution on mortality should be based on cohort studies. PMID- 11390323 TI - Vitamin supplement use and the risk of non-Hodgkin's lymphoma among women and men. AB - The authors examined use of individual supplements of vitamins A, C, and E only and multivitamins in relation to risk of non-Hodgkin's lymphoma in prospective cohorts of 88,410 women in the Nurses' Health Study (1980-1996), with 261 incident cases during 16 years of follow-up, and of 47,336 men in the Health Professionals Follow-Up Study (1986-1996), with 111 incident cases during 10 years of follow-up. Multivitamin use was associated with a higher risk of non Hodgkin's lymphoma among women but not among men; the multivariate relative risks for long-term duration (10 or more years) were 1.48 (95% confidence interval (CI): 1.01, 2.16) for women and 0.85 (95% CI: 0.45, 1.58) for men. The pooled multivariate relative risk from the two cohorts was 1.18 (95% CI: 0.70, 2.02). Use of individual supplements of vitamins A, C, and E only was not associated with risk among men. An increased risk associated with the use of individual supplements of vitamins A, C, and E only among women appeared to be secondary to the use of multivitamins by the same persons. Because an elevated risk among multivitamin users was not observed consistently in the two cohorts and the pooled data were not significant, the elevated risk among women may be the result of chance. PMID- 11390324 TI - Vitamin supplement use and fatal non-Hodgkin's lymphoma among US men and women. AB - The authors evaluated the association between use of individual supplements of vitamins A, C, and E only and multivitamins and fatal non-Hodgkin's lymphoma in a large prospective mortality study of US men and women. During 14 years of follow up (1982-1996), 1,571 non-Hodgkin's lymphoma deaths among 508,351 men and 1,398 non-Hodgkin's lymphoma deaths among 676,306 women were documented. Long-term regular use of individual supplements of vitamins A, C, and E only and multivitamins was unrelated to fatal non-Hodgkin's lymphoma among either men or women. The multivariate relative risks for men who used supplements for 10 or more years were 1.03 (95% confidence interval (CI): 0.54, 2.00) for vitamin A supplements, 1.04 (95% CI: 0.78, 1.39) for vitamin C supplements, 1.06 (95% CI: 0.74, 1.51) for vitamin E supplements, and 1.14 (95% CI: 0.92, 1.40) for multivitamins. The multivariate relative risks for women who used supplements for 10 or more years were 1.40 (95% CI: 0.77, 2.54) for vitamin A supplements, 1.19 (95% CI: 0.89, 1.60) for vitamin C supplements, 1.27 (95% CI: 0.87, 1.84) for vitamin E supplements, and 1.21 (95% CI: 0.98, 1.50) for multivitamins. All associations became weaker when vitamin supplements were mutually adjusted. These findings do not support an important relation between long-term regular use of individual supplements of vitamins A, C, and E only and multivitamins and fatal non-Hodgkin's lymphoma. PMID- 11390325 TI - Fracture history and risk of breast and endometrial cancer. AB - Fractures in postmenopausal women may serve as a surrogate measure of bone density, reflecting long-term lower estrogen levels, and lower estrogen levels appear to be inversely associated with breast and endometrial cancer. Breast cancer cases aged 50-79 years (n = 5,559) and endometrial cancer cases aged 40-79 years (n = 739) were enrolled in a US case-control study in 1992-1994 to evaluate the relation between fractures and risk of breast and endometrial cancer. Controls for the breast cancer analysis (n = 5,829) and the endometrial cancer analysis (n = 2,334) were randomly selected from population lists (driver's license and Medicare files). Information on fracture history and other risk factors was obtained by telephone interview. Compared with women without a fracture in the past 5 years, the odds ratios for women with a history of fracture were 0.80 (95% confidence interval (CI): 0.68, 0.94) for breast cancer and 0.59 (95% CI: 0.40, 0.89) for endometrial cancer. Height loss (> or =2.5 cm) and recent fracture history were associated with the lowest risk of breast cancer (odds ratio = 0.62, 95% CI: 0.46, 0.83) and endometrial cancer (odds ratio = 0.15, 95% CI: 0.05, 0.43). These data suggest that the endogenous hormonal factors associated with increased fracture risk are also related to decreased breast cancer risk and, more strongly, to endometrial cancer risk. PMID- 11390326 TI - Risk of late-stage breast cancer after a childbirth. AB - A pregnancy may lead to hormone-induced growth of breast tumors. The authors investigated whether women in the first years after childbirth had a higher incidence of breast cancer and, in particular, a higher incidence of late-stage tumors (i.e., a large tumor, nodal involvement, or histologic grading II + III). The study was based on a population-based cohort of 1.5 million Danish women born between 1935 and 1978. Between 1978 and 1994, 10,790 incident cases of breast cancer were identified in a nationwide cancer registry. Overall, uniparous and biparous mothers experienced a transient increased risk that did not appear to be attributable to delayed cancer diagnosis. The risk of being diagnosed with a tumor whose diameter was larger than 5 cm was, on average, 53% higher during the first 10 years after birth compared with later. The risk of tumors of less than 2 cm was not significantly associated with time since the latest birth. In conclusion, after a childbirth, mothers experience a transient increased risk of breast cancer and, in particular, a relatively high risk of late-stage disease. This finding suggests that pregnancy-related factors transiently induce a high growth rate in cells that are already malignant and stimulate new tumor growth. PMID- 11390327 TI - Birth outcome in relation to licorice consumption during pregnancy. AB - A role for glucocorticoids is suspected in the etiology of low birth weight. The authors tested whether maternal consumption of glycyrrhizin (an inhibitor of cortisol metabolism) in licorice affects birth weight in humans. A sample of 1,049 Finnish women and their healthy singleton infants was studied in 1998. Glycyrrhizin intake was calculated from detailed questionnaires on licorice consumption. Glycyrrhizin exposure was grouped into three levels: low (<250 mg/week; n = 751), moderate (250-499 mg/week; n = 145), and heavy (> or =500 mg/week; n = 110). Birth weight and gestational age (from ultrasound measurements) were obtained from hospital records. Babies with heavy exposure to glycyrrhizin were not significantly lighter at birth, but they were significantly more likely to be born earlier: The odds ratio for being born before 38 weeks' gestation was 2.5 (95% confidence interval: 1.1, 5.5; p = 0.03). Although the effect of heavy glycyrrhizin intake on mean duration of gestation was small (2.52 days) when expressed as an effect on the mean, this shift to the left of the distribution of duration of gestation was sufficient to double the risk of being born before 38 weeks. The association remained in multivariate analyses. In conclusion, heavy glycyrrhizin exposure during pregnancy did not significantly affect birth weight or maternal blood pressure, but it was significantly associated with lower gestational age. PMID- 11390328 TI - Steroids and risk of upper gastrointestinal complications. AB - Most antiinflammatory drugs have been associated with an increase in upper gastrointestinal complications. However, the literature on steroids is more limited than that on nonsteroidal antiinflammatory drugs (NSAIDs). To estimate the risk of upper gastrointestinal complications associated with use of steroids alone and in combination, a nested case-control analysis was conducted on the General Practice Research Database from the United KINGDOM: The authors identified 2,105 cases of upper gastrointestinal complications and 11,500 controls between 1993 and 1998. The adjusted odds ratios associated with current use of oral steroids were 1.8 (95% confidence interval (CI): 1.3, 2.4) for upper gastrointestinal complications overall and 2.4 (95% CI: 1.7, 3.4) for gastric and 1.2 (95% CI: 0.8, 1.9) for duodenal damage. Steroids were similarly associated with bleeding (odds ratio (OR) = 1.8; 95% CI: 1.3, 2.4) and perforations (OR = 1.6; 95% CI: 0.9, 3.1). Simultaneous use of steroids with low-medium and high NSAID doses, respectively, produced odds ratios of 4.0 (95% CI: 1.3, 12.0) and 12.7 (95% CI: 6.2, 26.1), compared with users of none. Whenever possible, antiinflammatory drugs should be given in monotherapy and at the lowest effective dose in order to reduce the risk of upper gastrointestinal complications. PMID- 11390329 TI - Serum levels of C-reactive protein are associated with obesity, weight gain, and hormone replacement therapy in healthy postmenopausal women. AB - The authors evaluated the cross-sectional and prospective associations between the serum concentration of C-reactive protein and measures of obesity and fat distribution, hormone replacement therapy (HRT) use, and serum sex hormones in postmenopausal women from the Healthy Women Study (Allegheny County, Pennsylvania, 1998). The authors tested the hypothesis that C-reactive protein levels would be higher among HRT users and among women with greater body mass index, waist circumference, or visceral fat. There were 207 women in the study who were > or =8 years postmenopausal (101 HRT users and 106 HRT nonusers). The median levels of C-reactive protein were 3.01 mg/liter in HRT users compared with 1.74 mg/liter in nonusers (p = 0.002). C-reactive protein levels were strongly positively correlated with measures of body size, fatness, fat distribution, and weight gain among HRT users and nonusers. C-reactive protein was also positively correlated with serum estrone levels (r(s) = 0.38) among HRT nonusers. The highest level of C-reactive protein was found among HRT users in the highest quartile of visceral fat (4.29 mg/liter) compared with women not on HRT and in the lowest quartile of visceral fat (0.96 mg/liter). The use of HRT and measures of overall body fatness are important correlates of C-reactive protein among postmenopausal women. PMID- 11390330 TI - Alcohol consumption with age: a cross-sectional and longitudinal study of the Atherosclerosis Risk in Communities (ARIC) study, 1987-1995. AB - Previous cross-sectional and longitudinal studies assessing the association between age and drinking are inconsistent. Evaluating 15,425 Black and White men and women from four communities, this study sought to determine whether there was a consistent relation between age and drinking in cross-sectional and longitudinal analyses and to determine change in drinking status and level of consumption (occasional, light to moderate, and heavier drinkers) at follow-up. Cross-sectional analyses of drinking were performed for Atherosclerosis Risk in Communities examinations 1 (1987-1989) and 3 (1993-1995). The changes in drinking status and level were determined for the 12,565 persons with information at both examinations. Prevalence of drinking was generally inversely associated with age in the cross-sectional analyses for all ethnic/gender groups, and drinking prevalence decreased over the 6 years of follow-up for all except Black women. Only among Black drinkers was younger age associated with a higher level of alcohol consumption in both cross-sectional and prospective analyses. Thus, whether drinking prevalence declines, the amount consumed by drinkers is decreased, or whether both factors contribute to the decrease appears to vary with ethnicity and gender. The change in drinking level was substantial with more than 40% of baseline drinkers reporting drinking cessation or a different level of consumption at follow-up. PMID- 11390331 TI - Risk factors for all-terrain vehicle injuries: a national case-control study. AB - A case-control study design was used to determine and quantify all-terrain vehicle (ATV) risk factors. The analysis was based on the results of two national probability surveys conducted in 1997: a survey of injured ATV drivers treated in hospital emergency departments and a survey of the general population of ATV users. Cases were drawn from the injury survey; controls (ATV drivers who had not been injured) were drawn from the user survey. Risk factors were quantified by means of a binary logistic regression analysis. After adjustment for covariates, injury risks were systematically related to a number of driver characteristics (age, gender, driving experience), driver use patterns (monthly driving times, recreational vs. nonrecreational use), and vehicle characteristics (number of wheels, engine size). The results of the analysis suggest that future safety efforts should focus on reducing child injuries, getting new drivers to participate in hands-on training programs, and encouraging consumers to dispose of the three-wheel ATVs still in use. PMID- 11390332 TI - Comparison of telephone sampling and area sampling: response rates and within household coverage. AB - Random digit dialing is used frequently in epidemiologic case-control studies to select population-based controls, even when both cases and controls are interviewed face-to-face. However, concerns persist about the potential biases of random digit dialing, particularly given its generally lower response rates. In an Atlanta, Georgia, case-control study of breast cancer among women aged 20-54 years, all of whom were interviewed face-to-face, two statistically independent control groups were compared: those obtained through random digit dialing (n = 652) and those obtained through area probability sampling (n = 640). The household screening rate was significantly higher for the area sample, by 5.5%. Interview response rates were comparable. The telephone sample estimated a significantly larger percentage (by approximately 7%) of households to have no age-eligible women. Both control groups, appropriately weighted, had characteristics similar to US Census demographic characteristics for Atlanta women, except that respondents in both control groups were more educated and more likely to be married. The authors conclude that households contacted through random digit dialing are somewhat less likely to participate in the household screening process, and if they are cooperative, some households may not disclose that age-eligible women reside therein. Investigators need to develop improved methods for screening and enumerating household members in random digit dialing surveys that target a specific subpopulation, such as women. PMID- 11390333 TI - Accuracy of self-reports of acquired immunodeficiency syndrome and acquired immunodeficiency syndrome-related conditions in women. AB - To investigate the validity of self-reported acquired immunodeficiency syndrome (AIDS) among women enrolled in a prospective study of human immunodeficiency virus (HIV) infection, the authors compared the self-reported occurrence of AIDS specific diagnoses with AIDS diagnoses documented by county AIDS surveillance registries. Also examined was the association between participant characteristics and the validity of self-reports. Among the 339 HIV-infected participants in the Northern California Women's Interagency HIV Study between October 1994 and September 1998, 217 reported having been given a diagnosis of AIDS. Of these 217 women, 157 (72%) were listed in the registry as having AIDS. Among the specific AIDS-related conditions reported by three or more women, the sensitivity was highest for tuberculosis (100%), CD4 cell count less than 200 (84%), Mycobacterium avium complex (73%), and Pneumocystis carinii pneumonia (69%), and the positive predictive value was highest for CD4 cell count less than 200 (75%). Among all reported AIDS diagnoses, the kappa statistic was highest for cryptococcosis (0.67) and CD4 cell count less than 200 (0.57). The only statistically significant participant characteristic associated with inaccurate reporting of an AIDS diagnosis was being a current cigarette smoker (adjusted odds ratio = 2.57, 95% confidence interval: 1.17, 5.64). Overall, self-reporting of any AIDS-related condition is fairly accurate, but there is great variability in the accuracy of specific conditions. PMID- 11390334 TI - Anatomic-electrophysiological correlations concerning the pathways for atrioventricular conduction. AB - The remarkable success of radiofrequency ablation in recent decades in curing atrioventricular nodal reentrant tachycardias has intensified efforts to provide a solid theoretical basis for understanding the mechanisms of atrioventricular transmission. These efforts, which were made by both anatomists and electrophysiologists, frequently resulted in seemingly controversial observations. Quantitatively and qualitatively, our understanding of the mysteries of propagation through the inhomogeneous and extremely complex atrioventricular conduction axis is much deeper than it was at the beginning of the past century. We must go back to the initial sources, nonetheless, in an attempt to provide a common ground for evaluating the morphological and electrophysiological principles of junctional arrhythmias. In this review, we provide an account of the initial descriptions, which still provide an appropriate foundation for interpreting recent electrophysiological findings. PMID- 11390335 TI - Glycemic control and heart failure among adult patients with diabetes. AB - BACKGROUND: Glycemic control is associated with microvascular events, but its effect on the risk of heart failure is not well understood. We examined the association between hemoglobin (Hb) A(Ic) and the risk of heart failure hospitalization and/or death in a population-based sample of adult patients with diabetes and assessed whether this association differed by patient sex, heart failure pathogenesis, and hypertension status. METHODS AND RESULTS: A cohort design was used with baseline between January 1, 1995, and June 30, 1996, and follow-up through December 31, 1997 (median 2.2 years). Participants were 25 958 men and 22 900 women with (predominantly type 2) diabetes, >/=19 years old, with no known history of heart failure. There were a total of 935 events (516 among men; 419 among women). After adjustment for age, sex, race/ethnicity, education level, cigarette smoking, alcohol consumption, hypertension, obesity, use of beta blockers and ACE inhibitors, type and duration of diabetes, and incidence of interim myocardial infarction, each 1% increase in Hb A(Ic) was associated with an 8% increased risk of heart failure (95% CI 5% to 12%). An Hb A(Ic) >/=10, relative to Hb A(Ic) <7, was associated with 1.56-fold (95% CI 1.26 to 1.93) greater risk of heart failure. Although the association was stronger in men than in women, no differences existed by heart failure pathogenesis or hypertension status. CONCLUSIONS: These results confirm previous evidence that poor glycemic control may be associated with an increased risk of heart failure among adult patients with diabetes. PMID- 11390336 TI - Low dietary folate intake is associated with an excess incidence of acute coronary events: The Kuopio Ischemic Heart Disease Risk Factor Study. AB - BACKGROUND: Although several prospective studies have shown that low folate intake and low circulating folate are associated with increased risk of coronary heart disease (CHD), the findings are inconsistent. METHODS AND RESULTS: We studied the associations of dietary intake of folate, vitamin B(6), and vitamin B(12) with the risk of acute coronary events in a prospective cohort study of 1980 Finnish men 42 to 60 years old examined in 1984 to 1989 in the Kuopio Ischemic Heart Disease Risk Factor Study. Nutrient intakes were assessed by 4-day food record. During an average follow-up time of 10 years, 199 acute coronary events occurred. In a Cox proportional hazards model adjusted for 21 conventional and nutritional CHD risk factors, men in the highest fifth of folate intake had a relative risk of acute coronary events of 0.45 (95% CI 0.25 to 0.81, P=0.008) compared with men in the lowest fifth. This association was stronger in nonsmokers and light alcohol users than in smokers and alcohol users. A high dietary intake of vitamin B(6) had no significant association and that of vitamin B(12) a weak association with a reduced risk of acute coronary events. CONCLUSIONS: The present work in CHD-free middle-aged men is the first prospective cohort study to observe a significant inverse association between quantitatively assessed moderate-to-high folate intakes and incidence of acute coronary events in men. Our findings provide further support in favor of a role of folate in the promotion of good cardiovascular health. PMID- 11390337 TI - Immunohistological changes in dilated cardiomyopathy induced by immunoadsorption therapy and subsequent immunoglobulin substitution. AB - BACKGROUND: Immunoadsorption (IA) and subsequent immunoglobulin (Ig) G substitution represent an additional therapeutic approach in dilated cardiomyopathy (DCM). It remains to be elucidated whether this treatment modulates myocardial inflammation, which is possibly a causal factor of ventricular dysfunction. METHODS AND RESULTS: From 25 DCM patients (EF <30%), 12 patients were randomized for IA therapy and subsequent IgG substitution at 1 month intervals until month 3. Before (<7 days) and after IA therapy, right ventricular biopsies were obtained from all patients. Biopsies were also obtained at intervals of 3 months from 13 patients without IA/IgG treatment (controls). IA/IgG treatment induced improvement in left ventricular ejection fraction from 21.3+/-1.7% (+/-SEM) to 27.0+/-1.3% (P<0.01 versus baseline/controls) and reduction of the beta-receptor autoantibody serum levels (P<0.01 versus baseline/controls). The number of CD3 cells decreased from 5.7+/-0.8 to 2.9+/-0.5 cells/mm(2) (P<0.01 versus baseline/controls). This decline was paralleled by a decrease in CD4 (P<0.01 versus baseline/controls) and CD8 (P<0.05 versus baseline/controls) lymphocytes. The number of leukocyte common antigen-positive cells (leukocytes) was reduced from 20.0+/-3.2 to 9.9+/-2.8 cells/mm(2) (P<0.01 versus baseline/P<0.05 versus controls). HLA class II expression decreased from 2.1+/-0.7% to 1.1+/-0.4% (P<0.05 versus controls/baseline). The number of immunopositive cells and the expression of HLA class II in controls remained stable. In both groups, the degree of fibrosis remained unchanged. CONCLUSIONS: IA and subsequent IgG substitution mitigate myocardial inflammation in DCM. PMID- 11390338 TI - Clinical and echocardiographic characteristics of papillary fibroelastomas: a retrospective and prospective study in 162 patients. AB - BACKGROUND: Cardiac papillary fibroelastoma (CPF) is a primary cardiac neoplasm that is increasingly detected by echocardiography. The clinical manifestations of this entity are not well described. METHODS AND RESULTS: In a 16-year period, we identified patients with CPF from our pathology and echocardiography databases. A total of 162 patients had pathologically confirmed CPF. Echocardiography was performed in 141 patients with 158 CPFs, and 48 patients had CPFs that were not visible by echocardiography (<0.2 cm), leaving an echocardiographic subgroup of 93 patients with 110 CPFs. An additional 45 patients with a presumed diagnosis of CPF were identified. The mean age of the patients was 60+/-16 years of age, and 46.1% were male. Echocardiographically, the mean size of the CPFs was 9+/-4.6 mm; 82.7% occurred on valves (aortic more than mitral), 43.6% were mobile, and 91.4% were single. During a follow-up period of 11+/-22 months, 23 of 26 patients with a prospective diagnosis of CPF that was confirmed by pathological examination had symptoms that could be attributable to embolization. In the group of 45 patients with a presumed diagnosis of CPF, 3 patients had symptoms that were likely due to embolization (incidence, 6.6%) during a follow-up period of 552+/-706 days. CONCLUSIONS: CPFs are generally small and single, occur most often on valvular surfaces, and may be mobile, resulting in embolization. Because of the potential for embolic events, symptomatic patients, patients undergoing cardiac surgery for other lesions, and those with highly mobile and large CPFs should be considered for surgical excision. PMID- 11390339 TI - Astroglial protein S-100 is an early and sensitive marker of hypoxic brain damage and outcome after cardiac arrest in humans. AB - BACKGROUND: The results of early conventional tests do not correlate with cerebral outcome after cardiac arrest. We investigated the serum levels of astroglial protein S-100 as an early marker of brain damage and outcome after cardiac arrest. METHODS AND RESULTS: In 66 patients undergoing cardiopulmonary resuscitation after nontraumatic cardiac arrest, blood samples for the evaluation of S-100 were drawn immediately after and 15, 30, 45, and 60 minutes; 2, 8, 24, 48, and 72 hours; and 7 days after initiation of cardiopulmonary resuscitation. Moreover, the serum levels of neuron-specific enolase were determined between 2 hours and 7 days. If patients survived for >48 hours, brain damage was assessed by a combination of neurological, cranial CT, and electrophysiological examinations. Overall, 343 blood samples were taken for the determination of S 100. Maximum S-100 levels within 2 hours after cardiac arrest were significantly higher in patients with documented brain damage (survivors and nonsurvivors, 3.70+/-0.77 microg/L) than in patients without brain damage (0.90+/-0.29 microg/L). Significant differences between these 2 groups were observed from 30 minutes until 7 days after cardiac arrest. In addition, the positive predictive value of the S-100 test at 24 hours for fatal outcome within 14 days was 87%, and the negative predictive value was 100% (P<0.001). With regard to neuron-specific enolase, significant differences between patients with documented brain damage and those with no brain damage were found at 24, 48, and 72 hours and 7 days. CONCLUSIONS: Astroglial protein S-100 is an early and sensitive marker of hypoxic brain damage and short-term outcome after cardiac arrest in humans. PMID- 11390340 TI - Unrecognized pulmonary venous desaturation early after Norwood palliation confounds Gp:Gs assessment and compromises oxygen delivery. AB - BACKGROUND: Hemodynamic stability after Norwood palliation often requires manipulation of pulmonary vascular resistance to alter the pulmonary-to-systemic blood flow ratio (Qp:Qs). Qp:Qs is often estimated from arterial saturation (SaO2), a practice based on 2 untested assumptions: constant systemic arteriovenous O2 difference and normal pulmonary venous saturation. METHODS AND RESULTS: In 12 patients early (/=0.5 mm were defined as atherosclerotic. A total of 2014 sites within 1477 segments in 574 coronary arteries (2.2 arteries per person) were analyzed. An atherosclerotic lesion was present in 136 patients, or 51.9%. The prevalence of atherosclerosis varied from 17% in individuals <20 years old to 85% in subjects >/=50 years old. In subjects with atherosclerosis, intimal thickness and area stenosis averaged 1.08+/-0.48 mm and 32.7+/-15.9%, respectively. For all age groups, the average intimal thickness was greater in men than women, although the prevalence of atherosclerosis was similar (52% in men and 51.7% in women). CONCLUSIONS: This study demonstrates that coronary atherosclerosis begins at a young age and that lesions are present in 1 of 6 teenagers. These findings suggest the need for intensive efforts at coronary disease prevention in young adults. PMID- 11390342 TI - Artificial neural network-based method of screening heart murmurs in children. AB - BACKGROUND: Early recognition of heart disease is an important goal in pediatrics. Efforts in developing an inexpensive screening device that can assist in the differentiation between innocent and pathological heart murmurs have met with limited success. Artificial neural networks (ANNs) are valuable tools used in complex pattern recognition and classification tasks. The aim of the present study was to train an ANN to distinguish between innocent and pathological murmurs effectively. METHODS AND RESULTS: Using an electronic stethoscope, heart sounds were recorded from 69 patients (37 pathological and 32 innocent murmurs). Sound samples were processed using digital signal analysis and fed into a custom ANN. With optimal settings, sensitivities and specificities of 100% were obtained on the data collected with the ANN classification system developed. For future unknowns, our results suggest the generalization would improve with better representation of all classes in the training data. CONCLUSION: We demonstrated that ANNs show significant potential in their use as an accurate diagnostic tool for the classification of heart sound data into innocent and pathological classes. This technology offers great promise for the development of a device for high-volume screening of children for heart disease. PMID- 11390343 TI - Homocysteine induces expression and secretion of monocyte chemoattractant protein 1 and interleukin-8 in human aortic endothelial cells: implications for vascular disease. AB - BACKGROUND: Proinflammatory cytokines play key roles in atherogenesis and disease progression. Because hyperhomocysteinemia is an independent risk factor for cardiovascular disease, we hypothesized that homocysteine could be atherogenic by altering the expression of specific cytokines in vascular endothelial cells. METHODS AND RESULTS: Northern blot and RNase protection assays showed that DL homocysteine induced mRNA expression of the proinflammatory cytokines monocyte chemoattractant protein-1 (MCP-1) and interleukin-8 (IL-8) in cultured human aortic endothelial cells (HAECs). Homocysteine had no effect on expression of other cytokines, namely tumor necrosis factor-alpha, granulocyte-macrophage colony-stimulating factor, interleukin-1beta, and transforming growth factor beta. MCP-1 mRNA expression increased 1 hour after homocysteine treatment, reached a maximum within 2 to 4 hours, and declined to basal levels over the next 24 hours. Induction of mRNA expression for both chemokines was observed with as little as 10 micromol/L DL-homocysteine, and maximal expression was achieved with 50 micromol/L DL-homocysteine. Homocysteine also triggered the release of MCP-1 and IL-8 protein from HAECs into the culture medium. The induction was specific for homocysteine, because equimolar concentrations of L-homocystine, L-cysteine, and L-methionine had no effect on mRNA levels and protein release. Furthermore, L homocysteine induced chemokine expression, but D-homocysteine did not, thus demonstrating enantiomeric specificity. The culture medium from homocysteine treated HAECs promoted chemotaxis in human peripheral blood monocytes and U937 cells. Anti-human recombinant MCP-1 antibody blocked the migration. CONCLUSIONS: Pathophysiological levels of L-homocysteine alter endothelial cell function by upregulating MCP-1 and IL-8 expression and secretion. This suggests that L homocysteine may contribute to the initiation and progression of vascular disease by promoting leukocyte recruitment. PMID- 11390344 TI - Comparative value of dobutamine and adenosine stress in the detection of coronary stenosis with myocardial contrast echocardiography. AB - BACKGROUND: Controversy continues as to whether adenosine or dobutamine is the superior pharmacological stress agent for myocardial contrast echocardiography (MCE). METHODS AND RESULTS: We compared real-time MCE refilling curves and wall thickening during adenosine and dobutamine stress in 14 open-chest dogs with left anterior descending and left circumflex coronary artery stenoses that reduced hyperemia by 40% to 60% and 70% to 90% (mild and severe non-flow-limiting stenosis, NFLS) and resting flow by 10% to 30% and 35% to 50% (mild and severe flow-limiting stenosis, FLS). MCE was performed with low-energy imaging during Optison infusion. After high-energy bubble destruction, time-intensity data from risk beds were fitted for an exponential function as y=A(1-e(-)(bt)), from which the rate of intensity increase (b) and maximal plateau intensity (A) were derived. Although severe NFLS and greater stenoses decreased b with both dobutamine and adenosine, with mild NFLS it was reduced in 58% of animals with dobutamine versus 8% with adenosine. The absolute decrease in b, however, was greater for adenosine than dobutamine with FLS. The A parameter was decreased with both adenosine and dobutamine only with the most severe FLS. Wall thickening was decreased with dobutamine in 33% of animals with severe NFLS and in all animals with any FLS; with adenosine, in all with severe FLS. CONCLUSIONS: Both dobutamine and adenosine significantly reduce MCE refilling rates in the setting of severe stenosis and in the absence of contractile abnormalities. Dobutamine decreases refilling rate and wall thickening at a less reduced flow grade than adenosine, but adenosine produces a greater magnitude of change than dobutamine. PMID- 11390345 TI - Cardiac-specific LIM protein FHL2 modifies the hypertrophic response to beta adrenergic stimulation. AB - BACKGROUND: A deficiency of muscle LIM protein results in dilated cardiomyopathy, but the function of other LIM proteins in the heart has not been assessed previously. We have characterized the expression and function of FHL2, a heart specific member of the LIM domain gene family. METHODS AND RESULTS: Expression of FHL2 mRNA and protein was examined by Northern blot, in situ hybridization, and Western blot analyses of fetal and adult mice. FHL2 transcripts are present at embryonic day (E) 7.5 within the cardiac crescent in a pattern that resembles that of Nkx2.5 mRNA. During later stages of cardiac development and in adult animals, FHL2 expression is localized to the myocardium and absent from endocardium, cardiac cushion, outflow tract, or coronary vasculature. The gene encoding FHL2 was disrupted by homologous recombination, and knockout mice devoid of FHL2 were found to undergo normal cardiovascular development. In the absence of FHL2, however, cardiac hypertrophy resulting from chronic infusion of isoproterenol is exaggerated (59% versus 20% increase in heart weight/body weight in FHL null versus wild-type mice; P<0.01). CONCLUSIONS: FHL2 is an early marker of cardiogenic cells and a cardiac-specific LIM protein in the adult. FHL2 is not required for normal cardiac development but modifies the hypertrophic response to beta-adrenergic stimulation. PMID- 11390346 TI - Increased expression of isoform 1 of the sarcoplasmic reticulum Ca(2+)-release channel in failing human heart. AB - BACKGROUND: The sarcoplasmic reticulum (SR) Ca(2+)-release channel plays a key role in the excitation-contraction coupling of cardiac myocytes. Because respective alterations have been reported in human heart failure, we investigated isoform expression of the SR Ca(2+)-release channel in human hearts from patients with terminal heart failure (dilated cardiomyopathy [DCM], n=8) and nonfailing organ donors (NF, n=8). METHODS AND RESULTS: Expression of mRNA of SR Ca(2+) release channel isoforms in isolated human cardiomyocytes and myocardial tissue was analyzed by reverse-transcription polymerase chain reaction. Protein expression was quantified in myocardial tissue with [(3)H]-ryanodine binding and with Western blots, expressed as densitometric units per microgram of protein (DU), and cellular localization was visualized with immunohistochemistry. We found mRNA expression of isoforms 1, 2, and 3 in cardiomyocytes and myocardial tissue both in NF and DCM. Total SR Ca(2+)-release channel protein expression in NF (B(max) 2.16+/-0.43 pmol/mg protein) and in DCM (B(max) 2.33+/-0.22 pmol/mg protein) myocardium was unchanged. Expression of isoform 1 of the SR Ca(2+) release channel was significantly (P=0.0037) increased in DCM myocardium (NF 1.97+/-0.25 versus DCM 3.37+/-0.31 DU), whereas protein expression of isoform 2 (NF 14.62+/-0.87 versus DCM 13.52+/-0.43 DU) and isoform 3 (NF 1.39+/-0.13 versus DCM 1.35+/-0.19 DU) was unchanged. All 3 isoforms of the protein could be localized in human ventricular myocytes with fluorescence immunohistochemistry. CONCLUSIONS: All 3 isoforms of the SR Ca(2+)-release channel were determined in human ventricular cardiomyocytes. Increased expression of isoform 1 of the SR Ca(2+)-release channel could contribute to impaired excitation-contraction coupling in human heart failure. PMID- 11390347 TI - Double-outlet right ventricle and overriding tricuspid valve reflect disturbances of looping, myocardialization, endocardial cushion differentiation, and apoptosis in TGF-beta(2)-knockout mice. AB - BACKGROUND: Transforming growth factor-beta(2) (TGF-beta(2)) is a member of a family of growth factors with the potential to modify multiple processes. Mice deficient in the TGF-beta(2) gene die around birth and show a variety of defects of different organs, including the heart. METHODS AND RESULTS: We studied the hearts of TGF-beta(2)-null mouse embryos from 11.5 to 18.5 days of gestation to analyze the types of defects and determine which processes of cardiac morphogenesis are affected by the absence of TGF-beta(2). Analysis of serial sections revealed malformations of the outflow tract (typically a double-outlet right ventricle) in 87.5%. There was 1 case of common arterial trunk. Abnormal thickening of the semilunar valves was seen in 4.2%. Associated malformations of the atrioventricular (AV) canal were found in 62.5% and were composed of perimembranous inlet ventricular septal defects (37.5%), AV valve thickening (33.3%), overriding tricuspid valve (25.0%), and complete AV septal defects (4.2%). Anomalies of the aorta and its branches were seen in 33.3%. Immunohistochemical staining showed failure of myocardialization of the mesenchyme of the atrial septum and the ventricular outflow tract as well as deficient valve differentiation. Morphometry documented this to be associated with absence of the normal decrease of total endocardial cushion volume in the older stages. Apoptosis in TGF-beta(2)-knockout mice was increased, although regional distribution was normal. CONCLUSIONS: TGF-beta(2)-knockout mice exhibited characteristic cardiovascular anomalies comparable to malformations seen in the human population. PMID- 11390348 TI - Liposome-mediated gene transfection of endothelial nitric oxide synthase reduces endothelial activation and leukocyte infiltration in transplanted hearts. AB - BACKGROUND: During cardiac ischemia-reperfusion injury, neutrophilic infiltration of the myocardium is mediated by adhesion molecule expression on activated coronary endothelium. Nitric oxide inhibits neutrophil adhesion to endothelium in vitro by blocking the nuclear factor (NF)-kappaB-dependent transcription of adhesion molecules. We investigated whether intraoperative gene delivery of endothelial nitric oxide synthase (eNOS) into donor hearts before transplantation would have a similar effect on an entire organ. METHODS AND RESULTS: In an allogeneic rabbit heart transplant model, liposomes complexed to the gene encoding eNOS were infused into the donor coronary circulation before transplantation. By 24 hours after transplantation, calcium-dependent nitrite production was significantly higher in eNOS-transfected donor hearts than in the 3 control groups: donor hearts transfected with empty plasmids alone, donor hearts treated with diluent only, and untransplanted native hearts. Intramyocardial neutrophil and T-lymphocyte populations were halved in eNOS transfected hearts compared with control donor hearts (P<0.05). Moreover, the prevalence of NF-kappaB activation in microvascular endothelial cells and surrounding cardiac myocytes as well as endothelial vascular cell adhesion molecule-1 and intracellular adhesion molecule-1 expression were all significantly reduced in eNOS-transfected hearts compared with control donor hearts (P<0.01). Without immunosuppression, eNOS-transfected hearts survived longer than controls. CONCLUSIONS: Intraoperative liposome-mediated gene delivery of eNOS to donor hearts can result in early gene expression sufficient to reduce ischemia-reperfusion injury by inhibiting NF-kappaB activation, adhesion molecule expression, and the early infiltration of leukocytes, all of which may improve graft survival. PMID- 11390349 TI - Therapeutic potential of nitric oxide synthase gene manipulation. PMID- 11390350 TI - Prinzmetal angina documented by transtelephonic electrocardiographic monitoring. PMID- 11390351 TI - Impending paradoxical embolism. PMID- 11390353 TI - Malignant glioma: genetics and biology of a grave matter. PMID- 11390355 TI - A Caenorhabditis elegans cohesion protein with functions in meiotic chromosome pairing and disjunction. AB - We have studied four Caenorhabditis elegans homologs of the Rad21/Scc1/Rec8 sister-chromatid cohesion protein family. Based on the RNAi phenotype and protein localization, it is concluded that one of them, W02A2.6p, is the likely worm ortholog of yeast Rec8p. The depletion of C. elegans W02A2.6p (called REC-8) by RNAi, induced univalent formation and splitting of chromosomes into sister chromatids at diakinesis. Chromosome synapsis at pachytene was defective, but primary homology recognition seemed unaffected, as a closer-than-random association of homologous fluorescence in situ hybridization (FISH) signals at leptotene/zygotene was observed. Depletion of REC-8 also induced chromosome fragmentation at diakinesis. We interpret these fragments as products of unrepaired meiotic double-stranded DNA breaks (DSBs), because fragmentation was suppressed in a spo-11 background. Thus, REC-8 seems to be required for successful repair of DSBs. The occurrence of DSBs in REC-8-depleted meiocytes suggests that DSB formation does not depend on homologous synapsis. Anti-REC-8 immunostaining decorated synaptonemal complexes (SCs) at pachytene and chromosomal axes in bivalents and univalents at diakinesis. Between metaphase I and metaphase II, REC-8 is partially lost from the chromosomes. The partial loss of REC-8 from chromosomes between metaphase I and metaphase II suggests that worm REC-8 might function similarly to yeast Rec8p. The loss of yeast Rec8p from chromosome arms at meiosis I and centromeres at meiosis II coordinates the disjunction of homologs and sister chromatids at the two meiotic divisions. PMID- 11390354 TI - The Drosophila Su(var)2-10 locus regulates chromosome structure and function and encodes a member of the PIAS protein family. AB - The conserved heterochromatic location of centromeres in higher eukaryotes suggests that intrinsic properties of heterochromatin are important for chromosome inheritance. Based on this hypothesis, mutations in Drosophila melanogaster that alter heterochromatin-induced gene silencing were tested for effects on chromosome inheritance. Here we describe the characterization of the Su(var)2-10 locus, initially identified as a Suppressor of Position-Effect Variegation. Su(var)2-10 is required for viability, and mutations cause both minichromosome and endogenous chromosome inheritance defects. Mitotic chromosomes are improperly condensed in mutants, and polytene chromosomes are structurally abnormal and disorganized in the nucleus. Su(var)2-10 encodes a member of the PIAS protein family, a group of highly conserved proteins that control diverse functions. SU(VAR)2-10 proteins colocalize with nuclear lamin in interphase, and little to no SU(VAR)2-10 is found on condensed mitotic chromosomes. SU(VAR)2-10 is present at some polytene chromosome telomeres, and FISH analyses in mutant polytene nuclei revealed defects in telomere clustering and telomere-nuclear lamina associations. We propose that Su(var2-10 controls multiple aspects of chromosome structure and function by establishing/maintaining chromosome organization in interphase nuclei. PMID- 11390356 TI - Pds1 phosphorylation in response to DNA damage is essential for its DNA damage checkpoint function. AB - In Saccharomyces cerevisiae, Pds1 is an anaphase inhibitor and plays an essential role in DNA damage and spindle checkpoint pathways. Pds1 is phosphorylated in response to DNA damage but not spindle disruption, indicating distinct mechanisms delaying anaphase entry. Phosphorylation of Pds1 is Mec1 and Chk1 dependent in vivo. Here, we show that Pds1 is phosphorylated at multiple sites in vivo in response to DNA damage by Chk1. Mutation of the Chk1 phosphorylation sites on Pds1 abolished most of its DNA damage-inducible phosphorylation and its checkpoint function, whereas its anaphase inhibitor functions and spindle checkpoint functions remain intact. Loss of Pds1 phosphorylation correlates with APC-dependent Pds1 destruction in response to DNA damage. We also show that APC(Cdc20) is active in preanaphase arrested cells after DNA damage. This suggests that Pds1 is stabilized by phosphorylation in response to DNA damage, but APC(Cdc20) activity is not altered. Our results indicate that phosphorylation of Pds1 by Chk1 is the key function of Chk1 required to prevent anaphase entry. PMID- 11390357 TI - In vivo mechanisms by which tumors producing thrombospondin 1 bypass its inhibitory effects. AB - Thrombospondin 1 (TSP1) is a multifunctional protein able to activate TGFbeta and to inhibit angiogenesis in vivo. Although usually thought of as an inhibitor of tumor growth, TSP1 may sometimes be present at high levels during tumor progression, suggesting that tumors can eventually overcome their anti-tumor effects. Using a tet-repressible expression system, we demonstrate that murine TSP1 delayed the onset of tumor growth when produced in the tumor bed by rat fibrosarcoma tumor cells or by stromal fibroblasts coinjected with unmodified C6 glioma tumor cells. Yet upon prolonged exposure to TSP1, tumors came to grow at the same rate in the presence as in the absence of TSP1 and transplantation experiments showed that they had become insensitive to inhibition by TSP1 in both syngeneic and immune compromised hosts. Tumor resistance to TSP1 developed as a result of the in vivo outgrowth of pre-existing tumor cell variants that (1) secreted increased amounts of angiogenic factors that counterbalanced the inhibitory effect of TSP1 on neovascularization and (2) grew more efficiently in the presence of TSP1-activated TGFbeta. These results indicate that prolonged and continuous local delivery of a single multifunctional angiogenesis inhibitor like TSP1 to fast-growing tumors can lead to tumor resistance in vivo by fostering the outgrowth of subpopulations that are a by-product of the genetic instability of the tumor cells themselves. PMID- 11390358 TI - TSC1 and TSC2 tumor suppressors antagonize insulin signaling in cell growth. AB - Tuberous sclerosis is a human disease caused by mutations in the TSC1 or the TSC2 tumor suppressor gene. Previous studies of a Drosophila TSC2 homolog suggested a role for the TSC genes in maintaining DNA content, with loss of TSC2 leading to polyploidy and increased cell size. We have isolated mutations in the Drosophila homolog of the TSC1 gene. We show that TSC1 and TSC2 form a complex and function in a common pathway to control cellular growth. Unlike previous studies, our work shows that TSC1(-) or TSC2(-) cells are diploid. We find that, strikingly, the heterozygosity of TSC1 or TSC2 is sufficient to rescue the lethality of loss-of function insulin receptor mutants. Further genetic analyses suggest that the TSC genes act in a parallel pathway that converges on the insulin pathway downstream from Akt. Taken together, our studies identified the TSC tumor suppressors as novel negative regulators of insulin signaling. PMID- 11390359 TI - Delta signaling from the germ line controls the proliferation and differentiation of the somatic follicle cells during Drosophila oogenesis. AB - The body axes of Drosophila are established during oogenesis through reciprocal interactions between the germ line cells and the somatic follicle cells that surround them. The Notch pathway is required at two stages in this process: first, for the migration of the follicle cells around the germ line cyst and, later, for the polarization of the anterior-posterior (A-P) axis of the oocyte. Its function in these events, however, has remained controversial. Using clonal analysis, we show that Notch signaling controls cell proliferation and differentiation in the whole follicular epithelium. Notch mutant follicle cells remain in a precursor state and fail to switch from the mitotic cell cycle to the endocycle. Furthermore, removal of Delta from the germ line produces an identical phenotype, showing that Delta signals from the germ cells to control the timing of follicle cell differentiation. This explains the axis formation defects in Notch mutants, which arise because undifferentiated posterior follicle cells cannot signal to polarize the oocyte. Delta also signals from the germ line to Notch in the soma earlier in oogenesis to control the differentiation of the polar and stalk follicle cells. The germ line therefore regulates the development of the follicle cells through two complementary signaling pathways: Gurken signals twice to control spatial patterning, whereas Delta signals twice to exert temporal control. PMID- 11390360 TI - Inhibition of early apoptotic events by Akt/PKB is dependent on the first committed step of glycolysis and mitochondrial hexokinase. AB - The serine/threonine kinase Akt/PKB is a major downstream effector of growth factor-mediated cell survival. Activated Akt, like Bcl-2 and Bcl-xL, prevents closure of a PT pore component, the voltage-dependent anion channel (VDAC); intracellular acidification; mitochondrial hyperpolarization; and the decline in oxidative phosphorylation that precedes cytochrome c release. However, unlike Bcl 2 and Bcl-xL, the ability of activated Akt to preserve mitochondrial integrity, and thereby inhibit apoptosis, requires glucose availability and is coupled to its metabolism. Hexokinases are known to bind to VDAC and directly couple intramitochondrial ATP synthesis to glucose metabolism. We provide evidence that such coupling serves as a downstream effector function for Akt. First, Akt increases mitochondria-associated hexokinase activity. Second, the antiapoptotic activity of Akt requires only the first committed step of glucose metabolism catalyzed by hexokinase. Finally, ectopic hexokinase expression mimics the ability of Akt to inhibit cytochrome c release and apoptosis. We therefore propose that Akt increases coupling of glucose metabolism to oxidative phosphorylation and regulates PT pore opening via the promotion of hexokinase VDAC interaction at the outer mitochondrial membrane. PMID- 11390361 TI - MKK7 is an essential component of the JNK signal transduction pathway activated by proinflammatory cytokines. AB - Mitogen-activated protein kinases (MAPK) are activated by phosphorylation on Thr and Tyr by MAPK kinases. Two MAPK kinases (MKK4 and MKK7) can activate the c-Jun NH(2)-terminal kinase (JNK) group of MAPK in vitro. JNK is phosphorylated preferentially on Tyr by MKK4 and on Thr by MKK7. Targeted gene-disruption studies in mice were performed to examine the role of MKK4 and MKK7 in vivo. Simultaneous disruption of the Mkk4 and Mkk7 genes was required to block JNK activation caused by exposure of cells to environmental stress. In contrast, disruption of the Mkk7 gene alone was sufficient to prevent JNK activation caused by proinflammatory cytokines. These data demonstrate that MKK4 and MKK7 serve different functions in the JNK signal transduction pathway. PMID- 11390362 TI - A mutation in the Gsk3-binding domain of zebrafish Masterblind/Axin1 leads to a fate transformation of telencephalon and eyes to diencephalon. AB - Zebrafish embryos homozygous for the masterblind (mbl) mutation exhibit a striking phenotype in which the eyes and telencephalon are reduced or absent and diencephalic fates expand to the front of the brain. Here we show that mbl(-/-) embryos carry an amino-acid change at a conserved site in the Wnt pathway scaffolding protein, Axin1. The amino-acid substitution present in the mbl allele abolishes the binding of Axin to Gsk3 and affects Tcf-dependent transcription. Therefore, Gsk3 activity may be decreased in mbl(-/-) embryos and in support of this possibility, overexpression of either wild-type Axin1 or Gsk3beta can restore eye and telencephalic fates to mbl(-/-) embryos. Our data reveal a crucial role for Axin1-dependent inhibition of the Wnt pathway in the early regional subdivision of the anterior neural plate into telencephalic, diencephalic, and eye-forming territories. PMID- 11390363 TI - Regulated protein degradation controls PKA function and cell-type differentiation in Dictyostelium. AB - Cullins function as scaffolds that, along with F-box/WD40-repeat-containing proteins, mediate the ubiquitination of proteins to target them for degradation by the proteasome. We have identified a cullin CulA that is required at several stages during Dictyostelium development. culA null cells are defective in inducing cell-type-specific gene expression and exhibit defects during aggregation, including reduced chemotaxis. PKA is an important regulator of Dictyostelium development. The levels of intracellular cAMP and PKA activity are controlled by the rate of synthesis of cAMP and its degradation by the cAMP specific phosphodiesterase RegA. We show that overexpression of the PKA catalytic subunit (PKAcat) rescues many of the culA null defects and those of cells lacking FbxA/ChtA, a previously described F-box/WD40-repeat-containing protein, suggesting CulA and FbxA proteins are involved in regulating PKA function. Whereas RegA protein levels drop as the multicellular organism forms in the wild type strain, they remain high in culA null and fbxA null cells. Although PKA can suppress the culA and fbxA null developmental phenotypes, it does not suppress the altered RegA degradation, suggesting that PKA lies downstream of RegA, CulA, and FbxA. Finally, we show that CulA, FbxA, and RegA are found in a complex in vivo, and formation of this complex is dependent on the MAP kinase ERK2, which is also required for PKA function. We propose that CulA and FbxA regulate multicellular development by targeting RegA for degradation via a pathway that requires ERK2 function, leading to an increase in cAMP and PKA activity. PMID- 11390364 TI - Two enhancers and one silencer located in the introns of regA control somatic cell differentiation in Volvox carteri. AB - The regA gene plays a central role in germ-soma differentiation of Volvox carteri by suppressing all reproductive functions in somatic cells. Here we show that the minimal promoter of regA consists of only 42 bp immediately upstream of the transcription start site, and that it contains no discernible regulatory elements. However, introns 3 and 5 are both required for regA expression in somatic cells, and intron 7 is essential for silencing regA in gonidia (asexual reproductive cells). A regA gene lacking intron 7 rescues the normal phenotype of mutant somatic cells, but also results in gonidia that reproduce only weakly and soon die out. The same phenotype is observed when a regA gene containing intron 7 is placed under control of a constitutive promoter, suggesting that the silencing activity of intron 7 is promoter specific. Intron 7 is unusual in that it contains a potential ORF that is in frame with exons 7 and 8, and some transcripts are produced in which intron 7 is retained. However, a regulatory role for the intron 7 translation product can be ruled out, because a construct in which intron 7 must be translated, and one in which it cannot be translated, both result in wild-type development of both cell types. Furthermore, intron 7 is unable to act in trans to silence regA, but is able to exert its normal effect when placed in a different location within the gene. Therefore, it appears that intron 7 functions in gonidia as a classical cell-type-specific and promoter specific enhancer, of the inhibitory type that is often referred to as a silencer. PMID- 11390365 TI - Release from tonic inhibition of T cell activation through transient displacement of C-terminal Src kinase (Csk) from lipid rafts. AB - In resting peripheral T cells, Csk is constitutively present in lipid rafts through an interaction with the Csk SH2-binding protein, PAG, also known as Cbp. Upon triggering of the T cell antigen receptor (TCR), PAG/Cbp is rapidly dephosphorylated leading to dissociation of Csk from lipid rafts. However, tyrosine phosphorylation of PAG/Cbp resumes after 3--5 min, at which time Csk reassociates with the rafts. Cells overexpressing a mutant Csk that lacks the catalytic domain, but displaces endogenous Csk from lipid rafts, have elevated basal levels of TCR-zeta-chain phosphorylation and spontaneous activation of an NFAT-AP1 reporter from the proximal interleukin-2 promoter as well as stronger and more sustained responses to TCR triggering than controls. We suggest that a transient release from Csk-mediated inhibition by displacement of Csk from lipid rafts is important for normal T cell activation. PMID- 11390366 TI - Golgi-localizing, gamma-adaptin ear homology domain, ADP-ribosylation factor binding (GGA) proteins interact with acidic dileucine sequences within the cytoplasmic domains of sorting receptors through their Vps27p/Hrs/STAM (VHS) domains. AB - GGA (Golgi-localizing, gamma-adaptin ear homology domain, ARF-binding) proteins are potential effectors of ADP-ribosylation factors, are associated with the trans-Golgi network (TGN), and are involved in protein transport from this compartment. By yeast two-hybrid screening and subsequent two-hybrid and pull down analyses, we have shown that GGA proteins, through their VHS (Vps27p/Hrs/STAM) domains, interact with acidic dileucine sequences found in the cytoplasmic domains of TGN-localized sorting receptors such as sortilin and mannose 6-phosphate receptor. A mutational analysis has revealed that a leucine pair and a cluster of acidic residues adjacent to the pair are mainly responsible for the interaction. A chimeric receptor with the sortilin cytoplasmic domain localizes to the TGN, whereas the chimeric receptor with a mutation at the leucine pair or the acidic cluster is mislocalized to punctate structures reminiscent of early endosomes. These results indicate that GGA proteins regulate the localization to or exit from the TGN of the sorting receptors. PMID- 11390368 TI - Identification of Ipaf, a human caspase-1-activating protein related to Apaf-1. AB - Procaspase-9 contains an NH2-terminal caspase-associated recruitment domain (CARD), which is essential for direct association with Apaf-1 and activation. Procaspase-1 also contains an NH2-terminal CARD domain, suggesting that its mechanism of activation, like that of procaspase-9, involves association with an Apaf-1-related molecule. Here we describe the identification of a human Apaf-1 related protein, named Ipaf that contains an NH2-terminal CARD domain, a central nucleotide-binding domain, and a COOH-terminal regulatory leucine-rich repeat domain (LRR). Ipaf associates directly and specifically with the CARD domain of procaspase-1 through CARD-CARD interaction. A constitutively active Ipaf lacking its COOH-terminal LRR domain can induce autocatalytic processing and activation of procaspase-1 and caspase-1-dependent apoptosis in transfected cells. Our results suggest that Ipaf is a specific and direct activator of procaspase-1 and could be involved in activation of caspase-1 in response to pro-inflammatory and apoptotic stimuli. PMID- 11390367 TI - Islet-brain1/JNK-interacting protein-1 is required for early embryogenesis in mice. AB - Islet-brain1/JNK-interacting protein-1 (IB1/JIP-1) is a scaffold protein that organizes the JNK, MKK7, and MLK1 to allow signaling specificity. Targeted disruption of the gene MAPK8IP1 encoding IB1/JIP-1 in mice led to embryonic death prior to blastocyst implantation. In culture, no IB1/JIP-1(-/-) embryos were identified indicating that accelerated cell death occurred during the first cell cycles. IB1/JIP-1 expression was detected in unfertilized oocytes, in spermatozoa, and in different stages of embryo development. Thus, despite the maternal and paternal transmission of the IB1/JIP-1 protein, early transcription of the MAPK8IP1 gene is required for the survival of the fertilized oocytes. PMID- 11390369 TI - A multiprotein complex that interacts with RNA polymerase II elongator. AB - A three-subunit Hap complex that interacts with the RNA polymerase II Elongator was isolated from yeast. Deletions of genes for two Hap subunits, HAP1 and HAP3, confer pGKL killer-insensitive and weak Elongator phenotypes. Preferential interaction of the Hap complex with free rather than RNA polymerase II-associated Elongator suggests a role in the regulation of Elongator activity. PMID- 11390370 TI - Ligand stimulation reduces platelet-derived growth factor beta-receptor susceptibility to tyrosine dephosphorylation. AB - Ligand binding to the platelet-derived growth factor (PDGF) beta-receptor leads to increased receptor tyrosine phosphorylation as a consequence of dimerization induced activation of the intrinsic receptor tyrosine kinase activity. In this study we asked whether ligand-stimulated PDGF beta-receptor tyrosine phosphorylation, to some extent, also involved reduced susceptibility to tyrosine dephosphorylation. To investigate this possibility we compared the sensitivity of ligand-stimulated and non-stimulated forms of tyrosine-phosphorylated PDGF beta receptors to dephosphorylation using various preparations containing protein tyrosine phosphatase activity. Ligand-stimulated or unstimulated tyrosine phosphorylated receptors were obtained after incubation of cells with pervanadate only or pervanadate, together with PDGF-BB, respectively. Dephosphorylation of receptors immobilized on wheat germ agglutinin-Sepharose, as well as of receptors in intact cell membranes, was investigated under conditions when rephosphorylation did not occur. As compared with unstimulated receptors the ligand-stimulated PDGF beta-receptors showed about 10-fold reduced sensitivity to dephosphorylation by cell membranes, a recombinant form of the catalytic domain of density-enhanced phosphatase-1, or recombinant protein-tyrosine phosphatase 1B. We conclude that ligand-stimulated forms of the PDGF beta-receptor display a reduced susceptibility to dephosphorylation. Our findings suggest a novel mechanism whereby ligand stimulation of PDGF beta-receptor, and possibly other tyrosine kinase receptors, leads to a net increase in receptor tyrosine phosphorylation. PMID- 11390371 TI - Functional coupling between secretory and cytosolic phospholipase A2 modulates tumor necrosis factor-alpha- and interleukin-1beta-induced NF-kappa B activation. AB - Tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta are potent activators of the transcription factor NF-kappaB, induced during inflammatory conditions. We have previously shown that both secretory and cytosolic phospholipase A(2) (PLA(2)) are involved in TNF-alpha- and IL-1beta-induced NF kappaB activation. In this study, we have addressed the mechanism of PLA(2) involvement with respect to downstream arachidonic acid (AA) metabolites and the functional coupling between PLA(2)s mediating NF-kappaB activation. We show that in addition to inhibitors of secretory and cytosolic PLA(2)s, 5-lipoxygenase inhibitors attenuate TNF-alpha- and IL-1beta-stimulated NF-kappaB activation. Exogenous addition of leukotriene B(4) (LTB(4)) restored NF-kappaB activation reduced by 5-lipoxygenase inhibitors or an LTB(4) receptor antagonist, thus identifying LTB(4) as a mediator in signaling to NF-kappaB. TNF-alpha- and IL 1beta-induced AA release from cellular membranes was accompanied by phosphorylation of cytosolic PLA(2). Inhibitors of secretory PLA(2) and of 5 lipoxygenase/LTB(4) functionality markedly reduced AA release and nearly completely abolished cytosolic PLA(2) phosphorylation. This demonstrates that secretory PLA(2), through 5-lipoxygenase metabolites, is an essential upstream regulator of cytosolic PLA(2) and AA release. Our results therefore suggest the existence of a functional link between secretory and cytosolic PLA(2) in cytokine activated keratinocytes, providing a molecular explanation for the participation of both secretory and cytosolic PLA(2) in arachidonic acid signaling and NF kappaB activation in response to proinflammatory cytokines. PMID- 11390372 TI - Specific binding of glucose-derivatized polymers to the asialoglycoprotein receptor of mouse primary hepatocytes. AB - In this study, we designed a novel amphiphilic poly-(p-N-vinylbenzyl-D glucuronamide) (PV6Gna) modified at the 6-OH position of glucose for hepatocyte recognition to address the mechanism of the interaction between mouse primary hepatocytes and the PV6Gna. PV6Gna bound to lectins specific for glucose but not galactose as did other glucose-derivatized polymers. However, hepatocyte adhesion onto the PV6Gna surface was inhibited in the presence of galactose and its analogues but not in the presence of glucose and its analogues. We also showed that hepatocyte adhesion to the PV6Gna surface was inhibited dose dependently by asialofetuin (ASF). Interactions between soluble PV6Gna and hepatocytes were inhibited by GalNAc, ASF, and EGTA in flow cytometry analysis using fluorescein isothiocyanate-conjugated PV6Gna. Hepatocyte adhesion to the PV6Gna surface was inhibited more effectively by GalNAc than by methyl beta-D-galactose. In flow cytometry analysis and cell adhesion assay, ASF competed for the inhibition of interaction between PV6Gna and hepatocytes 0.5-4 x 10(5)-fold more effectively than did GalNAc. These results demonstrate involvement of asialoglycoprotein receptors (ASGPRs) in the interaction between PV6Gna and hepatocytes. Furthermore, to clarify the mechanism of the interaction between glycopolymers modified at the 6-OH position of glucose and the hepatocyte, we prepared a gel particle containing 6-O-methacryloyl-d-glucose (PMglc) synthesized by an enzymatic method. ASGPRs could be detected using Western blot analysis following precipitation with PMglc in hepatocyte cell lysate. The precipitation of ASGPRs was inhibited in the presence of galactose, ASF, PV6Gna, and EGTA. The precipitation was inhibited more effectively by GalNAc than by methyl beta-d galactose. ASGPRs were rarely precipitated by PMglc in the cell lysate that had been treated with ASF-conjugated Sepharose. Taken together, we suggest that mouse primary hepatocytes adhere to the PV6Gna surface mediated by ASGPRs, which specifically interacted with the glycopolymers modified at the C-6 position of glucose. PMID- 11390373 TI - Dual action of eicosapentaenoic acid in hepatoma cells: up-regulation of metabolic action of insulin and inhibition of cell proliferation. AB - Exogenous administration of eicosapentaenoic acid (EPA) improves insulin sensitivity, but its precise mechanism remains unknown. Here we show that EPA stimulates the intracellular insulin signaling pathway in hepatoma cells. Exposure of these cells to EPA caused up-regulation of several insulin-induced activities including tyrosine phosphorylation of insulin receptor substrate-1, insulin receptor substrate-1-associated phosphatidylinositol 3-kinase, and its downstream target Akt kinase activity as well as down-regulation of gluconeogenesis. In contrast, EPA decreased mitogen-activated protein kinase activity and inhibited cell proliferation. These findings raise the possibility that EPA up-regulates metabolic action of insulin and inhibits cell growth in humans. PMID- 11390374 TI - Ligands of macrophage scavenger receptor induce cytokine expression via differential modulation of protein kinase signaling pathways. AB - Our previous works demonstrated that ligands of macrophage scavenger receptor (MSR) induce protein kinases (PKs) including protein-tyrosine kinase (PTK) and up regulate urokinase-type plasminogen activator expression (Hsu, H. Y., Hajjar, D. P., Khan, K. M., and Falcone, D. J. (1998) J. Biol. Chem. 273, 1240--1246). To continue to investigate MSR ligand-mediated signal transductions, we focus on ligands, oxidized low density lipoprotein (OxLDL), and fucoidan induction of the cytokines tumor necrosis factor-alpha (TNF) and interleukin 1 beta (IL-1). In brief, in murine macrophages J774A.1, OxLDL and fucoidan up-regulate TNF production; additionally, fucoidan but not OxLDL induces IL-1 secretion, prointerleukin 1 (proIL-1, precursor of IL-1) protein, and proIL-1 message. Simultaneously, fucoidan stimulates activity of interleukin 1-converting enzyme. We further investigate the molecular mechanism by which ligand binding-induced PK mediated mitogen-activated protein kinase (MAPK) in regulation of expression of proIL-1 and IL-1. Specifically, fucoidan stimulates activity of p21-activated kinase (PAK) and of the MAPKs extracellular signal-regulated kinase (ERK), c-Jun NH(2)-terminal kinase (JNK), and p38. Combined with PK inhibitors and genetic mutants of Rac1 and JNK in PK activity assays, Western blotting analyses, and IL 1 enzyme-linked immunosorbent assay, the role of individual PKs in the regulation of proIL-1/IL-1 was extensively dissected. Moreover, tyrosine phosphorylation of pp60Src as well as association between pp60Src and Hsp90 play important roles in fucoidan-induced proIL-1 expression. We are the first to establish two fucoidan mediated signaling pathways: PTK(Src)/Rac1/PAK/JNK and PTK(Src)/Rac1/PAK/p38, but not PTK/phospholipase C-gamma 1/PKC/MEK1/ERK, playing critical roles in proIL 1/IL-1 regulation. Our current results indicate and suggest a model for MSR ligands differentially modulating specific PK signal transduction pathways, which regulate atherogenesis-related inflammatory cytokines TNF and IL-1. PMID- 11390375 TI - Extracellular glycosaminoglycans modify cellular trafficking of lipoplexes and polyplexes. AB - It has been shown that extracellular glycosaminoglycans (GAGs) limit the gene transfer by cationic lipids and polymers. The purpose of this study was to clarify how interactions with anionic GAGs (hyaluronic acid and heparan sulfate) modify the cellular uptake and distribution of lipoplexes and polyplexes. Experiments on cellular DNA uptake and GFP reporter gene expression showed that decreased gene expression can rarely be explained by lower cellular uptake. In most cases, the cellular uptake is not changed by GAG binding to the lipoplexes or polyplexes. Reporter gene expression is decreased or blocked by heparan sulfate, but it is increased by hyaluronic acid; this suggests that intracellular factors are involved. Confocal microscopy experiments demonstrated that extracellular heparan sulfate and hyaluronic acid are taken into cells both with free and DNA-associated carriers. We conclude that extracellular GAGs may alter both the cellular uptake and the intracellular behavior of the DNA complexes. PMID- 11390376 TI - The binding interaction of HMG-1 with the TATA-binding protein/TATA complex. AB - High mobility protein-1 (HMG-1) has been shown to regulate transcription by RNA polymerase II. In the context that it acts as a transcriptional repressor, it binds to the TATA-binding protein (TBP) to form the HMG-1/TBP/TATA complex, which is proposed to inhibit the assembly of the preinitiation complex. By using electrophoretic mobility shift assays, we show that the acidic C-terminal domain of HMG-1 and the N terminus of human TBP are the domains that are essential for the formation of a stable HMG-1/TBP/TATA complex. HMG-1 binding increases the affinity of TBP for the TATA element by 20-fold, which is reflected in a significant stimulation of the rate of TBP binding, with little effect on the dissociation rate constant. In support of the binding target of HMG-1 being the N terminus of hTBP, the N-terminal polypeptide of human TBP competes with and inhibits HMG-1/TBP/TATA complex formation. Deletion of segments of the N terminus of human TBP was used to map the region(s) where HMG-1 binds. These findings indicate that interaction of HMG-1 with the Q-tract (amino acids 55-95) in hTBP is primarily responsible for stable complex formation. In addition, HMG-1 and the monoclonal antibody, 1C2, specific to the Q-tract, compete for the same site. Furthermore, calf thymus HMG-1 forms a stable complex with the TBP/TATA complex that contains TBP from either human or Drosophila but not yeast. This is again consistent with the importance of the Q-tract for this stable interaction and shows that the interaction extends over many species but does not include yeast TBP. PMID- 11390377 TI - Identification of a novel A20-binding inhibitor of nuclear factor-kappa B activation termed ABIN-2. AB - The nuclear factor kappaB (NF-kappaB) plays a central role in the regulation of genes implicated in immune responses, inflammatory processes, and apoptotic cell death. The zinc finger protein A20 is a cellular inhibitor of NF-kappaB activation by various stimuli and plays a critical role in terminating NF-kappaB responses. The underlying mechanism for NF-kappaB inhibition by A20 is still unknown. A20 has been shown to interact with several proteins including tumor necrosis factor (TNF) receptor-associated factors 2 and 6, as well as the inhibitory protein of kappaB kinase (IKK) gamma protein. Here we report the cloning and characterization of ABIN-2, a previously unknown protein that binds to the COOH-terminal zinc finger domain of A20. NF-kappaB activation induced by TNF and interleukin-1 is inhibited by overexpression of ABIN-2. The latter also inhibits NF-kappaB activation induced by overexpression of receptor-interacting protein or TNF receptor-associated factor 2. In contrast, NF-kappaB activation by overexpression of IKKbeta or direct activators of the IKK complex, such as Tax, cannot be inhibited by ABIN-2. These results indicate that ABIN-2 interferes with NF-kappaB activation upstream of the IKK complex and that it might contribute to the NF-kappaB-inhibitory function of A20. PMID- 11390378 TI - The Plasmodium falciparum bifunctional ornithine decarboxylase, S-adenosyl-L methionine decarboxylase, enables a well balanced polyamine synthesis without domain-domain interaction. AB - In the human malaria parasite Plasmodium falciparum (Pf), polyamines are synthesized by a bifunctional enzyme that possesses both ornithine decarboxylase (ODC) and S-adenosyl-l-methionine decarboxylase (AdoMetDC) activities. The mature enzyme consists of the heterotetrameric N-terminal AdoMetDC and the C-terminal dimeric ODC, which results in the formation of a heterotetrameric complex. For the native bifunctional protein a half-life longer than 2 h was determined, which is in contrast to the extreme short half-life of its mammalian monofunctional counterparts. The biological advantage of the plasmodial bifunctional ODC/AdoMetDC might be that the control of polyamine synthesis is achieved by only having to regulate the abundance and activity of one protein. An interesting feature in the regulation of the bifunctional protein is that putrescine inhibits PfODC activity approximately 10-fold more efficiently than the mammalian ODC activity, and in contrast to the mammalian AdoMetDC the activity of the PfAdoMetDC domain is not stimulated by the diamine. To analyze post-translational processing, polymerization, and domain-domain interactions, several mutant proteins were generated that have single mutations in either the PfODC or PfAdoMetDC domains. The exchange of amino acids essential for the activity of one domain had no effect on the enzyme activity of the other domain. Even prevention of the post-translational cleavage of the AdoMetDC domain or ODC dimerization and thus the interference with the folding of the protein hardly affected the activity of the partner domain. In addition, inhibition of the activity of the PfODC domain had no effect on the activity of the PfAdoMetDC domain and vice versa. These results demonstrate that no domain-domain interactions occur between the two enzymes of the bifunctional PfODC/AdoMetDC and that both enzymatic activities are operating as independent catalytic sites that do not affect each other. PMID- 11390379 TI - Interaction of the calcium-sensing receptor and filamin, a potential scaffolding protein. AB - In many cases, the biologic responses of cells to extracellular signals and the specificity of the responses cannot be explained solely on the basis of the interactions of known signaling proteins. Recently, scaffolding and adaptor proteins have been identified that organize signaling proteins in cells and that contribute to the nature and specificity of signaling pathways. In an effort to identify proteins that might organize the signaling system(s) activated by the extracellular Ca(2+) receptor (CaR), we used a bait construct representing the intracellular C terminus of the human CaR and the yeast two hybrid system to screen a human kidney cDNA library. We identified a clone representing the C terminal 1042 amino acids (aa) of the cytoskeletal protein filamin (ABP-280). Analysis of truncation and deletion constructs of the CaR C terminus and the filamin cDNA clone demonstrated that the CaR and filamin interact via regions containing aa 907-997 of the CaR C terminus and aa 1566-1875 of filamin. Interaction of the two proteins in mammalian HEK-293 cells was demonstrated by co immunoprecipitation and colocalization of them using immunofluorescence microscopy. The functional importance of their interaction was documented by transiently expressing the CaR in M2 melanoma cells that lack filamin, or in A7 melanoma cells that stably express filamin, and demonstrating that the CaR activated ERK only in the presence of filamin. Co-expression of the CaR with a peptide derived from the region of the CaR C terminus that interacts with filamin reduced the ability of the CaR to activate p42ERK in a dose-dependent manner, but did not inhibit the ability of the ET(A) receptor to activate ERK. The fact that filamin interacts with the CaR and other cell signaling proteins including mitogen-activated protein kinases and small GTPases, indicates that it may act as a scaffolding protein to organize cell signaling systems involving the CaR. PMID- 11390380 TI - Filamin-A binds to the carboxyl-terminal tail of the calcium-sensing receptor, an interaction that participates in CaR-mediated activation of mitogen-activated protein kinase. AB - The G protein-coupled, extracellular calcium-sensing receptor (CaR) regulates parathyroid hormone secretion and parathyroid cellular proliferation as well as the functions of diverse other cell types. The CaR resides in caveolae-plasma membrane microdomains containing receptors and associated signaling molecules that are thought to serve as cellular "message centers." An additional mechanism for coordinating cellular signaling is the presence of scaffold proteins that bind and organize components of signal transduction cascades. With the use of the yeast two-hybrid system, we identified filamin-A (an actin-cross-linking, putative scaffold protein that binds mitogen-activated protein kinase (MAPK) components activated by the CaR) as an intracellular binding partner of the CaR's carboxyl (COOH)-terminal tail. A direct interaction of the two proteins was confirmed by an in vitro binding assay. Moreover, confocal microscopy combined with two color immunofluorescence showed co-localization of the CaR and filamin-A within parathyroid cells as well as HEK-293 cells stably transfected with the CaR. Deletion mapping localized the sites of interaction between the two proteins to a stretch of 60 amino acid residues within the distal portion of the CaR's COOH-terminal tail and domains 14 and 15 in filamin-A, respectively. Finally, introducing the portion of filamin-A interacting with the CaR into CaR transfected HEK-293 cells using protein transduction with a His-tagged, Tat filamin-A fusion protein nearly abolished CaR-mediated activation of ERK1/2 MAPK but had no effect on ERK1/2 activity stimulated by ADP. Therefore, the binding of the CaR's COOH-terminal tail to filamin-A may contribute to its localization in caveolae, link it to the actin-based cytoskeleton, and participate in CaR mediated activation of MAPK. PMID- 11390381 TI - Roles of glucitol in the GutR-mediated transcription activation process in Bacillus subtilis: glucitol induces GutR to change its conformation and to bind ATP. AB - GutR is a 95-kDa glucitol-dependent transcription activator that mediates the expression of the Bacillus subtilis glucitol operon. Glucitol allows GutR to bind tightly to its binding site located upstream of the gut promoter. In this study, a second functional role of glucitol is identified. Glucitol induces GutR to change its conformation and triggers GutR to bind ATP efficiently. After sequential binding of glucitol and ATP to GutR, GutR adopts a new conformation by forming a compact structure that is resistant to trypsin digestion. Under this condition, the ATP.glucitiol.GutR complex can dissociate slowly from the gutR binding site (t(12) = 274 min). Interestingly, if ATP in the ATP.glucitiol.GutR complex is replaced by ADP, GutR adopts another conformation and can dissociate from the gutR-binding site even faster (t(12) = 82 min). In all these GutR-DNA binding studies in the presence of different ligands (glucitol, ATP, or ADP), only the off-rate is affected. The vital role of ATP in the GutR-mediated transcription activation process is reflected by the poor transcription from the gut promoter with GutR(D285A) which has a mutation in the motif B of the putative ATP-binding site. A working model for this transcription activation process is presented. PMID- 11390382 TI - Kruppel-like factor 4 (gut-enriched Kruppel-like factor) inhibits cell proliferation by blocking G1/S progression of the cell cycle. AB - Kruppel-like factor 4 (KLF4) is an epithelial cell-enriched, zinc finger containing transcription factor, the expression of which is associated with growth arrest. Previous studies show that constitutive expression of KLF4 inhibits DNA synthesis but the manner by which KLF4 exerts this effect is unclear. In the present study, we developed a system in which expression of KLF4 is controlled by a promoter that is induced upon treatment of cells containing the receptors for the insect hormone, ecdysone, with ponasterone A, an ecdysone analogue. The rate of proliferation of a stably transfected colon cancer cell line, RKO, was significantly decreased following addition of ponasterone A when compared with untreated cells. Flow cytometric analyses indicated that the inducible expression of KLF4 caused a block in the G(1)/S phase of the cell cycle. A similar block was observed when ecdysone receptor-containing RKO cells were infected with a replication-defective recombinant adenovirus containing an inducible KLF4 and treated with ponasterone A. Results of these studies provide evidence that the inhibitory effect of KLF4 on cell proliferation is mainly exerted at the G(1)/S boundary of the cell cycle. PMID- 11390383 TI - Conditional coupling of leading-strand and lagging-strand DNA synthesis at bacteriophage T4 replication forks. AB - Eight proteins encoded by bacteriophage T4 are required for the replicative synthesis of the leading and lagging strands of T4 DNA. We show here that active T4 replication forks, which catalyze the coordinated synthesis of leading and lagging strands, remain stable in the face of dilution provided that the gp44/62 clamp loader, the gp45 sliding clamp, and the gp32 ssDNA-binding protein are present at sufficient levels after dilution. If any of these accessory proteins is omitted from the dilution mixture, uncoordinated DNA synthesis occurs, and/or large Okazaki fragments are formed. Thus, the accessory proteins must be recruited from solution for each round of initiation of lagging-strand synthesis. A modified bacteriophage T7 DNA polymerase (Sequenase) can replace the T4 DNA polymerase for leading-strand synthesis but not for well coordinated lagging strand synthesis. Although T4 DNA polymerase has been reported to self-associate, gel-exclusion chromatography displays it as a monomer in solution in the absence of DNA. It forms no stable holoenzyme complex in solution with the accessory proteins or with the gp41-gp61 helicase-primase. Instead, template DNA is required for the assembly of the T4 replication complex, which then catalyzes coordinated synthesis of leading and lagging strands in a conditionally coupled manner. PMID- 11390384 TI - Phosphorylation of p42/44(MAPK) by various signal transduction pathways activates cytosolic phospholipase A(2) to variable degrees. AB - Arachidonic acid has been implicated to play a role in physiological and pathophysiological processes and is selectively released by the 85-kDa cytosolic phospholipase A(2) (cPLA(2)). The activity of cPLA(2) is regulated by calcium, translocating the enzyme to its substrate, and by phosphorylation by a mitogen activated protein kinase (MAPK) family member and a MAPK-activated protein kinase. In this study, the signal transduction pathways in growth factor-induced phosphorylation of p42/44(MAPK) and cPLA(2) activation were investigated in Her14 fibroblasts. p42/44(MAPK) in response to epidermal growth factor was not only phosphorylated via the Raf-MEK pathway but mainly through protein kinase C (PKC) or a related or unrelated kinase in which the phosphorylated p42/44(MAPK) corresponded with cPLA(2) activity. Serum-induced phosphorylation of p42/44(MAPK) also corresponded with cPLA(2) activity but is predominantly mediated via Raf-MEK and partly through PKC or a related or unrelated kinase. In contrast, activation of PKC by phorbol ester did not result in increased cPLA(2) activity, while p42/44(MAPK) is phosphorylated, mainly via Raf-MEK and through MEK. Moreover, p42/44(MAPK) phosphorylation is present in quiescent and proliferating cells, and p42/44(MAPK) is entirely phosphorylated via Raf-MEK, but it only corresponds to cPLA(2) activity in the former cells. Collectively, these data show that p42/44(MAPK) in proliferating, quiescent, and stimulated cells is phosphorylated by various signal transduction pathways, suggesting the activation of different populations of p42/44(MAPK) and cPLA(2). PMID- 11390385 TI - Cyclophilin a binds to peroxiredoxins and activates its peroxidase activity. AB - Six distinct peroxiredoxin (Prx) proteins (Prx I-VI) from distinct genes have been identified in mammalian tissues. Prxs are members of a group of peroxidases that have conserved reactive cysteine residue(s) in the active site(s). An immediate physiological electron donor for the peroxidase catalysis for five Prx proteins (Prx I-V) has been identified as thioredoxin (Trx), but that for Prx VI (1-Cys Prx) is still unclear. To identify an immediate electron donor and a binding protein for Prx VI, we performed a Prx VI protein overlay assay. A 20-kDa binding protein was identified by the Prx VI protein overlay assay with flow through fractions from a High-Q column with rat lung crude extracts. Using matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) and MS-Fit, we identified the 20-kDa Prx VI-binding protein as a cyclophilin A (CyP-A). The binding of recombinant human CyP-A (hCyP-A) to Prx VI was confirmed by using the hCyP-A protein overlay assay and Western immunoblot analysis with hCyP-A-specific antibodies. hCyP-A enhanced the antioxidant activity of Prx VI, as well as the other known mammalian Prx isotypes. hCyP-A supported antioxidant activity of Prx II and Prx VI both against thiol (dithiothreitol)-containing metal-catalyzed oxidation (MCO) systems and ascorbate-containing MCO systems. Prx II was reduced by hCyP-A without help from any other reductant, and the reduction was cyclosporin A-independent. These results strongly suggest that CyP-A not only binds to Prx proteins but also supports its peroxidase activity as an immediate electron donor. In addition, Cys(115) and Cys(161) of hCyP-A were found to be involved in the activation and the reduction of Prx. PMID- 11390386 TI - Tyrosine 36 plays a critical role in the interaction of the AB loop of tissue inhibitor of metalloproteinases-2 with matrix metalloproteinase-14. AB - The tissue inhibitor of metalloproteinases-2 (TIMP-2) is potentially an important inhibitor of all known matrix metalloproteinases (MMPs). However, it has been shown to undergo specific interactions with both MMP-2 (gelatinase A) and MMP-14 (MT1-MMP), and it has been proposed that these three proteins function as a cell surface-based activation cascade for matrix metalloproteinases and as a focus of proteolytic activity. In this study, we have carried out mutagenesis and kinetic analyses to examine the unique interactions between the AB loop of TIMP-2 and MMP 14. The results demonstrate that the major binding contribution of the AB loop is due solely to residue Tyr-36 at the tip of the hairpin. From this work, we propose that TIMP-2 may be engineered to abrogate MMP-14 binding, whereas its binding properties for other MMPs, including MMP-2, are maintained. Mutants of TIMP-2 with more directed specificity may be of use in gene therapeutic approaches to human disease. PMID- 11390387 TI - Functional analyses of Bph-Tod hybrid dioxygenase, which exhibits high degradation activity toward trichloroethylene. AB - Biphenyl dioxygenase (BphDox) in Pseudomonas pseudoalcaligenes KF707 is a multicomponent enzyme consisting of an iron-sulfur protein (ISP) that is composed of alpha (BphA1) and beta (BphA2) subunits, a ferredoxin (FD(BphA3)), and a ferredoxin reductase (FDR(BphA4)). A recombinant Escherichia coli strain expressing hybrid Dox that had replaced BphA1 with TodC1 (alpha subunit of toluene dioxygenase (TolDox) of Pseudomonas putida) exhibited high activity toward trichloroethylene (TCE) (Furukawa, K., Hirose, J., Hayashida, S., and Nakamura, K. (1994) J. Bacteriol. 176, 2121-2123). In this study, ISP, FD, and FDR were purified and characterized. Reconstitution of the dioxygenase components consisting of purified ISP(TodC1BphA2), FD(BphA3), and FDR(BphA4) exhibited oxygenation activities toward biphenyl, toluene, and TCE. Native polyacrylamide gel electrophoresis followed by the Ferguson plot analyses demonstrated that ISP(TodC1BphA2) and ISP(BphA1A2) were present as heterohexamers, whereas ISP(TodC1C2) was present as a heterotetramer. The molecular activity (k(0)) of the hybrid Dox for TCE was 4.1 min(-1), which is comparable to that of TolDox. The K(m) value of the hybrid Dox for TCE was 130 microm, which was lower than 250 microm for TolDox. These results suggest that the alpha subunit of ISP is crucial for the determination of substrate specificity and that the change in the alpha subunit conformation of ISP from alpha(2)beta(2) to alpha(3)beta(3) results in the acquisition of higher affinity to TCE, which may lead to high TCE degradation activity. PMID- 11390388 TI - Cyclopentenone prostaglandins of the J series inhibit the ubiquitin isopeptidase activity of the proteasome pathway. AB - Electrophilic eicosanoids of the J series, with their distinctive cross conjugated alpha,beta-unsaturated ketone, inactivate genetically wild type tumor suppressor p53 in a manner analogous to prostaglandins of the A series. Like the prostaglandins of the A series, prostaglandins of the J series have a structural determinant (endocyclic cyclopentenone) that confers the ability to impair the conformation, the phosphorylation, and the transcriptional activity of the p53 tumor suppressor with equivalent potency and efficacy. However, J series prostaglandins have a unique structural determinant (exocyclic alpha,beta unsaturated ketone) that confers unique efficacy as an apoptotic agonist. In seeking to understand how J series prostaglandins cause apoptosis despite their inactivation of p53, we discovered that they inhibit the ubiquitin isopeptidase activity of the proteasome pathway. In this regard, J series prostaglandins were more efficacious inhibitors than representative members of the A, B, or E series prostaglandins. Disruption of the proteasome pathway with proteasome inhibitors can cause apoptosis independently of p53. Therefore, this finding helps reconcile the p53 transcriptional independence of apoptosis caused by Delta12-prostaglandin J(2). This discovery represents a novel mechanism for proteasome pathway inhibition in intact cells. Furthermore, it identifies isopeptidases as novel targets for the development of antineoplastic agents. PMID- 11390389 TI - c-Abl tyrosine kinase binds and phosphorylates phospholipid scramblase 1. AB - Phospholipid scramblase 1 (PLSCR1) is a plasma membrane protein that has been proposed to play a role in the transbilayer movement of plasma membrane phospholipids. PLSCR1 contains multiple proline-rich motifs resembling Src homology 3 (SH3) domain-binding sites. An initial screen against 13 different SH3 domains revealed a marked specificity of PLSCR1 for binding to the Abl SH3 domain. Binding between intracellular PLSCR1 and c-Abl was demonstrated by co immunoprecipitation of both proteins from several cell lines. Deletion of the proline-rich segment in PLSCR1 (residues 1--118) abolished its binding to the Abl SH3 domain. PLSCR1 was Tyr-phosphorylated by c-Abl in vitro. Phosphorylation was abolished by mutation of Tyr residues Tyr(69)/Tyr(74) within the tandem repeat sequence (68)VYNQPVYNQP(77) of PLSCR1, implying that these residues are the likely sites of phosphorylation. Cellular PLSCR1 was found to be constitutively Tyr-phosphorylated in several cell lines. The Tyr phosphorylation of PLSCR1 was increased upon overexpression of c-Abl and significantly reduced either upon cell treatment with the Abl kinase inhibitor STI571, or in Abl-/- mouse fibroblasts, suggesting that cellular PLSCR1 is a normal substrate of c-Abl. Cell treatment with the DNA-damaging agent cisplatin activated c-Abl kinase and increased Tyr phosphorylation of PLSCR1. The cisplatin-induced phosphorylation of PLSCR1 was inhibited by STI571 and was not observed in Abl-/- fibroblasts. These findings indicate that c-Abl binds and phosphorylates PLSCR1, and raise the possibility that an interaction between c-Abl and plasma membrane PLSCR1 might contribute to the cellular response to genotoxic stress. PMID- 11390390 TI - Evolution of transglutaminase genes: identification of a transglutaminase gene cluster on human chromosome 15q15. Structure of the gene encoding transglutaminase X and a novel gene family member, transglutaminase Z. AB - We isolated and characterized the gene encoding human transglutaminase (TG)(X) (TGM5) and mapped it to the 15q15.2 region of chromosome 15 by fluorescence in situ hybridization. The gene consists of 13 exons separated by 12 introns and spans about 35 kilobases. Further sequence analysis and mapping showed that this locus contained three transglutaminase genes arranged in tandem: EPB42 (band 4.2 protein), TGM5, and a novel gene (TGM7). A full-length cDNA for the novel transglutaminase (TG(Z)) was obtained by anchored polymerase chain reaction. The deduced amino acid sequence encoded a protein with 710 amino acids and a molecular mass of 80 kDa. Northern blotting showed that the three genes are differentially expressed in human tissues. Band 4.2 protein expression was associated with hematopoiesis, whereas TG(X) and TG(Z) showed widespread expression in different tissues. Interestingly, the chromosomal segment containing the human TGM5, TGM7, and EPB42 genes and the segment containing the genes encoding TG(C),TG(E), and another novel gene (TGM6) on chromosome 20q11 are in mouse all found on distal chromosome 2 as determined by radiation hybrid mapping. This finding suggests that in evolution these six genes arose from local duplication of a single gene and subsequent redistribution to two distinct chromosomes in the human genome. PMID- 11390391 TI - The catalytic activities of the bifunctional Azotobacter vinelandii mannuronan C 5-epimerase and alginate lyase AlgE7 probably originate from the same active site in the enzyme. AB - The Azotobacter vinelandii genome encodes a family of seven secreted Ca(2+) dependent epimerases (AlgE1--7) catalyzing the polymer level epimerization of beta-D-mannuronic acid (M) to alpha-L-guluronic acid (G) in the commercially important polysaccharide alginate. AlgE1--7 are composed of two types of protein modules, A and R, and the A-modules have previously been found to be sufficient for epimerization. AlgE7 is both an epimerase and an alginase, and here we show that the lyase activity is Ca(2+)-dependent and also responds similarly to the epimerases in the presence of other divalent cations. The AlgE7 lyase degraded M rich alginates and a relatively G-rich alginate from the brown algae Macrocystis pyrifera most effectively, producing oligomers of 4 (mannuronan) to 7 units. The sequences cleaved were mainly G/MM and/or G/GM. Since G-moieties dominated at the reducing ends even when mannuronan was used as substrate, the AlgE7 epimerase probably stimulates the lyase pathway, indicating a complex interplay between the two activities. A truncated form of AlgE1 (AlgE1-1) was converted to a combined epimerase and lyase by replacing the 5'-798 base pairs in the algE1-1 gene with the corresponding A-module-encoding DNA sequence from algE7. Furthermore, substitution of an aspartic acid residue at position 152 with glycine in AlgE7A eliminated almost all of both the lyase and epimerase activities. Epimerization and lyase activity are believed to be mechanistically related, and the results reported here strongly support this hypothesis by suggesting that the same enzymatic site can catalyze both reactions. PMID- 11390392 TI - Carbohydrate recognition site of interleukin-2 in relation to cell proliferation. AB - Interleukin-2 (IL-2) is a cytokine with important roles in the immune system. IL 2 initially binds a high mannose-type glycan and a specific peptide sequence of the IL-2 receptor alpha-subunit and sequentially forms a high affinity complex of IL-2.IL-2 receptor alpha-, beta-, and gamma-subunits. This formation induces cellular signaling and cell proliferation (Fukushima, K., and Yamashita, K. (2001) J. Biol. Chem. 276, 7351-7356). To determine the carbohydrate-binding site of IL-2, we prepared wild-type and point-mutated (35)S-IL-2 by an in vitro transcription and translation method. We found that wild-type (35)S-IL-2 tends to form a dimer spontaneously, and the dimeric form has both carbohydrate recognition activity and cell proliferation activity. Moreover, substitution of Asn-26 in IL-2 with Gln or Asp conserved the dimeric form and affected the carbohydrate recognition activities in correspondence with the cell proliferation activities, suggesting that Asn-26 in IL-2 is involved in the carbohydrate recognition site. These results suggest that the carbohydrate recognition of IL-2 dimer triggers formation of high affinity complex (IL-2.IL-2Ralpha, -beta, gamma)(2), and the hetero-octamer stimulates IL-2-dependent T-cell proliferation by intensifying cellular signaling. PMID- 11390393 TI - Escherichia coli poly(A)-binding proteins that interact with components of degradosomes or impede RNA decay mediated by polynucleotide phosphorylase and RNase E. AB - The multifunctional ribonuclease RNase E and the 3'-exonuclease polynucleotide phosphorylase (PNPase) are major components of an Escherichia coli ribonucleolytic "machine" that has been termed the RNA degradosome. Previous work has shown that poly(A) additions to the 3' ends of RNA substrates affect RNA degradation by both of these enzymes. To better understand the mechanism(s) by which poly(A) tails can modulate ribonuclease action, we used selective binding in 1 m salt to identify E. coli proteins that interact at high affinity with poly(A) tracts. We report here that CspE, a member of a family of RNA-binding "cold shock" proteins, and S1, an essential component of the 30 S ribosomal subunit, are poly(A)-binding proteins that interact functionally and physically, respectively, with degradosome ribonucleases. We show that purified CspE impedes poly(A)-mediated 3' to 5' exonucleolytic decay by PNPase by interfering with its digestion through the poly(A) tail and also inhibits both internal cleavage and poly(A) tail removal by RNase E. The ribosomal protein S1, which is known to interact with sequences at the 5' ends of mRNA molecules during the initiation of translation, can bind to both RNase E and PNPase, but in contrast to CspE, did not affect the ribonucleolytic actions of these enzymes. Our findings raise the prospect that E. coli proteins that bind to poly(A) tails may link the functions of degradosomes and ribosomes. PMID- 11390394 TI - Kinetic investigation of chemokine truncation by CD26/dipeptidyl peptidase IV reveals a striking selectivity within the chemokine family. AB - Chemokines coordinate many aspects of leukocyte migration. As chemoattractants they play an important role in the innate and acquired immune response. There is good experimental evidence that N-terminal truncation by secreted or cell surface proteases is a way of modulating chemokine action. The localization of CD26/dipeptidyl peptidase IV on cell surfaces and in biological fluids, its primary specificity, and the type of naturally occurring truncated chemokines are consistent with such a function. We determined the steady-state catalytic parameters for a relevant selection of chemokines (CCL3b, CCL5, CCL11, CCL22, CXCL9, CXCL10, CXCL11, and CXCL12) previously reported to alter their chemotactic behavior due to CD26/dipeptidyl peptidase IV-catalyzed truncation. The results reveal a striking selectivity for stromal cell-derived factor-1alpha (CXCL12) and macrophage-derived chemokine (CCL22). The kinetic parameters support the hypothesis that CD26/dipeptidyl peptidase IV contributes to the degradation of certain chemokines in vivo. The data not only provide insight into the selectivity of the enzyme for specific chemokines, but they also contribute to the general understanding of CD26/dipeptidyl peptidase IV secondary substrate specificity. PMID- 11390395 TI - Requirement for high mobility group protein HMGI-C interaction with STAT3 inhibitor PIAS3 in repression of alpha-subunit of epithelial Na+ channel (alpha ENaC) transcription by Ras activation in salivary epithelial cells. AB - Previously, we have demonstrated that oxidative stress or Ras/ERK activation leads to the transcriptional repression of alpha-subunit of epithelial Na(+) channel (ENaC) in lung and salivary epithelial cells. Here, we further investigated the coordinated molecular mechanisms by which alpha-ENaC expression is regulated. Using both stable and transient transfection assays, we demonstrate that the overexpression of high mobility group protein I-C (HMGI-C), a Ras/ERK inducible HMG-I family member, represses glucocorticoid receptor (GR)/dexamethasone (Dex)-stimulated alpha-ENaC/reporter activity in salivary epithelial cells. Northern analyses further confirm that the expression of endogenous alpha-ENaC gene in salivary Pa-4 cells is suppressed by an ectopic HMGI-C overexpression. Through yeast two-hybrid screening and co immunoprecipitation assays from eukaryotic cells, we also discovered the interaction between HMGI-C and PIAS3 (protein inhibitor of activated STAT3 (signal transducer and activator of transcription 3)). A low level of ectopically expressed PIAS3 cooperatively inhibits GR/Dex-dependent alpha-ENaC transcription in the presence of HMGI-C. Reciprocally, HMGI-C expression also coordinately enhances PIAS3-mediated repression of STAT3-dependent transactivation. Moreover, overexpression of antisense HMGI-C construct is capable of reversing the repression mediated by Ras V12 on GR- and STAT3-dependent transcriptional activation. Together, our results demonstrate that Ras/ERK-mediated induction of HMGI-C is required to effectively repress GR/Dex-stimulated transcription of alpha-ENaC gene and STAT3-mediated transactivation. These findings delineate a network of inhibitory signaling pathways that converge on HMGI-C.PIAS3 complex, causally associating Ras/ERK activation with the repression of both GR and STAT3 signaling pathways in salivary epithelial cells. PMID- 11390396 TI - The (I/Y)XGG motif of adenovirus DNA polymerase affects template DNA binding and the transition from initiation to elongation. AB - Adenovirus DNA polymerase (Ad pol) is a eukaryotic-type DNA polymerase involved in the catalysis of protein-primed initiation as well as DNA polymerization. The functional significance of the (I/Y)XGG motif, highly conserved among eukaryotic type DNA polymerases, was analyzed in Ad pol by site-directed mutagenesis of four conserved amino acids. All mutant polymerases could bind primer-template DNA efficiently but were impaired in binding duplex DNA. Three mutant polymerases required higher nucleotide concentrations for effective polymerization and showed higher exonuclease activity on double-stranded DNA. These observations suggest a local destabilization of DNA substrate at the polymerase active site. In agreement with this, the mutant polymerases showed reduced initiation activity and increased K(m)(app) for the initiating nucleotide, dCMP. Interestingly, one mutant polymerase, while capable of elongating on the primer-template DNA, failed to elongate after protein priming. Further investigation of this mutant polymerase showed that polymerization activity decreased after each polymerization step and ceased completely after formation of the precursor terminal protein-trinucleotide (pTP-CAT) initiation intermediate. Our results suggest that residues in the conserved motif (I/Y)XGG in Ad pol are involved in binding the template strand in the polymerase active site and play an important role in the transition from initiation to elongation. PMID- 11390397 TI - Ultrastructure and function of dimeric, soluble intercellular adhesion molecule-1 (ICAM-1). AB - Previous studies have demonstrated dimerization of intercellular adhesion molecule-1 (ICAM-1) on the cell surface and suggested a role for immunoglobulin superfamily domain 5 and/or the transmembrane domain in mediating such dimerization. Crystallization studies suggest that domain 1 may also mediate dimerization. ICAM-1 binds through domain 1 to the I domain of the integrin alpha(L)beta(2) (lymphocyte function-associated antigen 1). Soluble C-terminally dimerized ICAM-1 was made by replacing the transmembrane and cytoplasmic domains with an alpha-helical coiled coil. Electron microscopy revealed C-terminal dimers that were straight, slightly bent, and sometimes U-shaped. A small number of apparently closed ring-like dimers and W-shaped tetramers were found. To capture ICAM-1 dimerized at the crystallographically defined dimer interface in domain 1, cysteines were introduced into this interface. Several of these mutations resulted in the formation of soluble disulfide-bonded ICAM-1 dimers (domain 1 dimers). Combining a domain 1 cysteine mutation with the C-terminal dimers (domain 1/C-terminal dimers) resulted in significant amounts of both closed ring like dimers and W-shaped tetramers. Surface plasmon resonance studies showed that all of the dimeric forms of ICAM-1 (domain 1, C-terminal, and domain 1/C-terminal dimers) bound similarly to the integrin alpha(L)beta(2) I domain, with affinities approximately 1.5--3-fold greater than that of monomeric ICAM-1. These studies demonstrate that ICAM-1 can form at least three different topologies and that dimerization at domain 1 does not interfere with binding in domain 1 to alpha(L)beta(2). PMID- 11390398 TI - Recombinational and mutagenic repair of psoralen interstrand cross-links in Saccharomyces cerevisiae. AB - Psoralen photoreacts with DNA to form interstrand cross-links, which can be repaired by both nonmutagenic nucleotide excision repair and recombinational repair pathways and by mutagenic pathways. In the yeast Saccharomyces cerevisiae, psoralen cross-links are processed by nucleotide excision repair to form double strand breaks (DSBs). In yeast, DSBs are repaired primarily by homologous recombination, predicting that cross-link and DSB repair should induce similar recombination end points. We compared psoralen cross-link, psoralen monoadduct, and DSB repair using plasmid substrates with site-specific lesions and measured the patterns of gene conversion, crossing over, and targeted mutation. Psoralen cross-links induced both recombination and mutations, whereas DSBs induced only recombination, and monoadducts were neither recombinogenic nor mutagenic. Although the cross-link- and DSB-induced patterns of plasmid integration and gene conversion were similar in most respects, they showed opposite asymmetries in their unidirectional conversion tracts: primarily upstream from the damage site for cross-links but downstream for DSBs. Cross-links induced targeted mutations in 5% of the repaired plasmids; all were base substitutions, primarily T --> C transitions. The major pathway of psoralen cross-link repair in yeast is error free and involves the formation of DSB intermediates followed by homologous recombination. A fraction of the cross-links enter an error-prone pathway, resulting in mutations at the damage site. PMID- 11390399 TI - Hormonal control of insulin-like growth factor I gene transcription in human osteoblasts: dual actions of cAMP-dependent protein kinase on CCAAT/enhancer binding protein delta. AB - Insulin-like growth factor-I (IGF-I) is essential for somatic growth and promotes bone cell replication and differentiation. IGF-I production by rat osteoblasts is stimulated by activation of cAMP-dependent protein kinase (PKA). In this report, we define two interacting PKA-regulated pathways that control IGF-I gene transcription in cultured human osteoblasts. Stimulation of cAMP led to a 12-fold increase in IGF-I mRNA and enhanced IGF-I promoter activity through a DNA response element termed HS3D and the transcription factor CCAAT/enhancer-binding protein delta (C/EBPdelta). Under basal conditions, C/EBPdelta was found in osteoblast nuclei but was transcriptionally silent. Treatment with the PKA inhibitor H-89 caused redistribution of C/EBPdelta to the cytoplasm. After hormone treatment, the catalytic subunit of PKA accumulated in osteoblast nuclei. Inhibition of active PKA with targeted nuclear expression of PKA inhibitor had no effect on the subcellular location of C/EBPdelta but prevented hormone-induced IGF-I gene activation, while cytoplasmic PKA inhibitor additionally caused the removal of C/EBPdelta from the nucleus. These results show that IGF-I gene expression is controlled in human osteoblasts by two PKA-dependent pathways. Cytoplasmic PKA mediates nuclear localization of C/EBPdelta under basal conditions, and nuclear PKA stimulates its transcriptional activity upon hormone treatment. Both mechanisms are indirect, since PKA failed to phosphorylate human C/EBPdelta in vitro. PMID- 11390400 TI - Facilitated transcription through the nucleosome at high ionic strength occurs via a histone octamer transfer mechanism. AB - The rate of transcription through the nucleosome, the fine structure of the nucleosomal barrier, and the fate of the nucleosome during transcription at different salt concentrations were analyzed using linear 227-base pair mononucleosomal templates containing a uniquely positioned nucleosome core. At lower ionic strength (30 mm NaCl), the nucleosome constitutes a strong barrier for SP6 RNA polymerase. At higher ionic strength (330 mm NaCl), the rates of transcription on nucleosomal and histone-free DNA templates are very similar. At both higher and lower ionic strengths, the complete histone octamer is transferred over the same distance by fundamentally similar mechanisms. The data indicate that even at the rate of transcription characteristic of histone-free DNA, the transfer intermediates can be formed quite efficiently. This suggests possible mechanisms that could facilitate transcription through the nucleosome at physiological ionic strength. PMID- 11390401 TI - Analysis of the V(D)J recombination efficiency at lymphoid chromosomal translocation breakpoints. AB - Chromosomal translocations and deletions are among the major events that initiate neoplasia. For lymphoid chromosomal translocations, misrecognition by the RAG (recombination activating gene) complex of V(D)J recombination is one contributing factor that has long been proposed. The chromosomal translocations involving LMO2 (t(11;14)(p13;q11)), Ttg-1 (t(11;14)(p15;q11)), and Hox11 (t(10;14)(q24;q11)) are among the clearest examples in which it appears that a D or J segment has synapsed with an adventitious heptamer/nonamer at a gene outside of one of the antigen receptor loci. The interstitial deletion at 1p32 involving SIL (SCL-interrupting locus)/SCL (stem cell leukemia) is a case involving two non V(D)J sites that have been suggested to be V(D)J recombination mistakes. Here we have used our human extrachromosomal substrate assay to formally test the hypothesis that these regions are V(D)J recombination misrecognition sites and, more importantly, to quantify their efficiency as V(D)J recombination targets within the cell. We find that the LMO2 fragile site functions as a 12-signal at an efficiency that is only 27-fold lower than that of a consensus 12-signal. The Ttg-1 site functions as a 23-signal at an efficiency 530-fold lower than that of a consensus 23-signal. Hox11 failed to undergo recombination as a 12- or 23 signal and was at least 20,000-fold less efficient than consensus signals. SIL has been predicted to function as a 12-signal and SCL as a 23-signal. However, we find that SIL actually functions as a 23-signal. These results provide a formal demonstration that certain chromosomal fragile sites can serve as RAG complex targets, and they determine whether these sites function as 12- versus 23 signals. These results quantify one of the three major factors that determine the frequency of these translocations in T-cell acute lymphocytic leukemia. PMID- 11390402 TI - FADD-deficient T cells exhibit a disaccord in regulation of the cell cycle machinery. AB - FADD is an adapter protein that was originally isolated as a transducer of apoptotic signals for death domain-containing receptors. However, FADD-deficient mice are embryonic lethal and FADD(-/-) T cells developed from FADD(-/-) embryonic stem cells in the RAG-1(-/-) hosts lack the full potential to proliferate when stimulated through their T-cell receptor complex, suggesting that FADD protein might play a dualistic role in mediating not only cell death signaling but other non-apoptotic cellular pathways as well. Here we show that a substantial number of freshly isolated FADD(-/-) peripheral T cells are cycling but are defective in their co-stimulatory response when stimulated. Analysis of several cell cycle proteins shows normal down-regulation of p27 inhibitor but increased levels of p21, decreased levels of cyclin D2, and constitutive activation of several cyclin-dependent kinases in activated T cells. These data suggest that FADD is involved in the regulation of cell cycle machinery in T lymphocytes. PMID- 11390403 TI - Photosynthetic water oxidation in cytochrome b(559) mutants containing a disrupted heme-binding pocket. AB - The role of cytochrome b(559) in photosynthetic oxygen evolution has been investigated in three chloroplast mutants of Chlamydomonas reinhardtii, in which one of the two histidine axial ligands to the heme, provided by the alpha subunit, has been replaced by the residues methionine, tyrosine, and glutamine. Photosystem two complexes functional for oxygen evolution could be assembled in the methionine and tyrosine mutants up to approximately 15% of wild type levels, whereas no complexes with oxygen evolution activity could be detected in the glutamine mutant. PSII supercomplexes isolated from the tyrosine and methionine mutants were as active as wild type in terms of light-saturated rates of oxygen evolution but in contrast to wild type contained no bound heme despite the presence of the alpha subunit. Oxygen evolution in the tyrosine and methionine mutants was, however, more sensitive to photoinactivation than the WT. Overall, these data establish unambiguously that a redox role for the heme of cytochrome b(559) is not required for photosynthetic oxygen evolution. Instead, our data provide new evidence of a role for cytochrome b(559) in the protection of the photosystem two complex in vivo. PMID- 11390404 TI - CCC1 is a transporter that mediates vacuolar iron storage in yeast. AB - The budding yeast Saccharomyces cerevisiae can grow for generations in the absence of exogenous iron, indicating a capacity to store intracellular iron. As cells can accumulate iron by endocytosis we studied iron metabolism in yeast that were defective in endocytosis. We demonstrated that endocytosis-defective yeast (Delta end4) can store iron in the vacuole, indicating a transfer of iron from the cytosol to the vacuole. Using several different criteria we demonstrated that CCC1 encodes a transporter that effects the accumulation of iron and Mn(2+) in vacuoles. Overexpression of CCC1, which is localized to the vacuole, lowers cytosolic iron and increases vacuolar iron content. Conversely, deletion of CCC1 results in decreased vacuolar iron content and decreased iron stores, which affect cytosolic iron levels and cell growth. Furthermore Delta ccc1 cells show increased sensitivity to external iron. The sensitivity to iron is exacerbated by ectopic expression of the iron transporter FET4. These results indicate that yeast can store iron in the vacuole and that CCC1 is involved in the transfer of iron from the cytosol to the vacuole. PMID- 11390405 TI - Palmitoylation-dependent control of degradation, life span, and membrane expression of the CCR5 receptor. AB - We have shown that the chemokine and HIV receptor CCR5 is palmitoylated on a cluster of cysteine residues located at the boundary between the seventh transmembrane region and the cytoplasmic tail. Single or combined substitutions of the three cysteines (Cys-321, Cys-323, and Cys-324) or incubation of wild-type CCR5-transfected cells with the palmitic acid analog 2-bromopalmitate prevented palmitoylation of the receptor. Moreover, failure of CCR5 to be palmitoylated resulted in both accumulation in intracellular stores and a profound decrease of membrane expression of the receptor. Upon metabolic labeling, kinetic experiments showed that the half-life of palmitoylation-deficient CCR5 is profoundly decreased. Bafilomycin A1, but not a specific proteasome inhibitor, prevented early degradation of palmitoylation-deficient CCR5 and promoted its accumulation in lysosomal compartments. Although membrane expression of the CCR5 mutant was diminished, the molecules reaching the membrane were still able to interact efficiently with the chemokine ligand MIP1 beta and remained able to function as HIV co-receptors. Thus we conclude that palmitoylation controls CCR5 expression through regulation of the life span of this receptor. PMID- 11390406 TI - Function of Escherichia coli biotin carboxylase requires catalytic activity of both subunits of the homodimer. AB - Biotin carboxylase catalyzes the ATP-dependent carboxylation of biotin and is one component of the multienzyme complex acetyl-CoA carboxylase that catalyzes the first committed step in fatty acid synthesis. The Escherichia coli biotin carboxylase is readily isolated from the other components of the acetyl-CoA carboxylase complex such that enzymatic activity is retained. The three dimensional structure of biotin carboxylase, determined by x-ray crystallography, demonstrated that the enzyme is a homodimer consisting of two active sites in which each subunit contains a complete active site. To understand how each subunit contributes to the overall function of biotin carboxylase, we made hybrid molecules in which one subunit had a wild-type active site, and the other subunit contained an active site mutation known to significantly affect the activity of the enzyme. One of the two genes encoded a poly-histidine tag at its N terminus, whereas the other gene had an N-terminal FLAG epitope tag. The two genes were assembled into a mini-operon that was induced to give high level expression of both enzymes. "Hybrid" dimers composed of one subunit with a wild-type active site and a second subunit having a mutant active site were obtained by sequential chromatographic steps on columns of immobilized nickel chelate and anti-FLAG affinity matrices. In vitro kinetic studies of biotin carboxylase dimers in which both subunits were wild type revealed that the presence of the N-terminal tags did not alter the activity of the enzyme. However, kinetic assays of hybrid dimer biotin carboxylase molecules in which one subunit had an active site mutation (R292A, N290A, K238Q, or E288K) and the other subunit had a wild-type active site resulted in 39-, 28-, 94-, and 285-fold decreases in the activity of these enzymes, respectively. The dominant negative effects of these mutant subunits were also detected in vivo by monitoring the rate of fatty acid biosynthesis by [(14)C]acetate labeling of cellular lipids. Expression of the mutant biotin carboxylase genes from an inducible arabinose promoter resulted in a significantly reduced rate of fatty acid synthesis relative to the same strain that expressed the wild type gene. Thus, both the in vitro and in vivo data indicate that both subunits of biotin carboxylase are required for activity and that the two subunits must be in communication during enzyme function. PMID- 11390407 TI - Activation of the hexosamine pathway leads to deterioration of pancreatic beta cell function through the induction of oxidative stress. AB - It is known well that activation of the hexosamine pathway causes insulin resistance, but how this activation influences pancreatic beta-cell function remains unclear. In this study, we found that in isolated rat islets adenovirus mediated overexpression of glutamine:fructose-6-phosphate amidotransferase (GFAT), the first and rate-limiting enzyme of the hexosamine pathway, leads to deterioration of beta-cell function, which is similar to that found in diabetes. Overexpression of GFAT or treatment with glucosamine results in impaired glucose stimulated insulin secretion and reduction in the expression levels of several beta-cell specific genes (insulin, GLUT2, and glucokinase). Additionally, the DNA binding activity of PDX-1, an important transcription factor for these three genes, was markedly reduced. These phenomena were not mimicked by the induction of O-linked glycosylation with an inhibitor of O-GlcNAcase, PUGNAc. It was also found that glucosamine increases hydrogen peroxide levels and that several hexosamine pathway-mediated changes were suppressed by treatment with the antioxidant N-acetyl-l-cysteine. In conclusion, activation of the hexosamine pathway leads to deterioration of beta-cell function through the induction of oxidative stress rather than O-linked glycosylation. Thus, the hexosamine pathway may contribute to the deterioration of beta-cell function found in diabetes. PMID- 11390408 TI - The hCds1 (Chk2)-FHA domain is essential for a chain of phosphorylation events on hCds1 that is induced by ionizing radiation. AB - hCds1 (Chk2) is an evolutionarily conserved kinase that functions in DNA damage response and cell cycle checkpoint. The Cds1 family of kinases are activated by a family of large phosphatidylinositol 3-kinase-like kinases. In humans, ataxia telangiectasia-mutated (ATM) and ataxia-telangiectasia and Rad3-related kinases activate hCds1 by phosphorylating Thr(68) . hCds1 and Cds1-related kinases contain the FHA (forkhead-associated) domain, which appears to be important for integrating the DNA damage signal. It is not known how ATM phosphorylation activates hCds1 function and whether the phosphorylation is linked to the FHA. Here, we demonstrate that the hCds1-FHA domain is essential for Thr(68) phosphorylation. Thr(68) phosphorylation, in turn, is required for ionizing radiation-induced autophosphorylation of two amino acid residues in hCds1, Thr(383) and Thr(387). These two amino acid residues, located in the activation loop of hCds1, are conserved in hCds1-related kinases and are essential for hCds1 activity. Thus, the hCds1-FHA domain mediates a chain of phosphorylation events on hCds1, which includes phosphorylation by ATM and hCds1 autophosphorylation, in response to DNA damage. PMID- 11390409 TI - The position of the alpha and beta subunits in a single chain variant of human chorionic gonadotropin affects the heterodimeric interaction of the subunits and receptor-binding epitopes. AB - The glycoprotein hormone family represents a class of heterodimers, which include the placental hormone human chorionic gonadotropin (CG) and the anterior pituitary hormones follitropin, lutropin, and thyrotropin. They are composed of common alpha subunit and a hormone-specific beta subunit. Based on the CG crystal structure, it was suggested that the quaternary subunit interactions are crucial for biological activity. However, recent observations using single chain glycoprotein hormone analogs, where the beta and alpha subunits are linked (NH(2) CGbeta-alpha; CGbetaalpha orientation), implied that the heterodimeric-like quaternary configuration is not a prerequisite for receptor binding/signal transduction. To study the heterodimeric alignment of the two subunit domains in a single chain and its role in the intracellular behavior and biological action of the hormone, a single chain CG variant was constructed in which the carboxyl terminus of alpha was fused to the CGbeta amino terminus (NH(2)-alpha-CGbeta; alphaCGbeta orientation). The secretion rate of alphaCGbeta from transfected Chinese hamster ovary cells was less than that seen for CGbetaalpha. The alphaCGbeta tether was not recognized by dimer-specific monoclonal antibodies and did not bind to lutropin/CG receptor. To define if one or both subunit domains were modified in alphaCGbeta, it was co-transfected with a monomeric alpha or CGbeta gene. In each case, alphaCGbeta/alpha and alphaCGbeta/CGbeta complexes were formed indicating that CG dimer-specific epitopes were established. The alphaCGbeta/alpha complex bound to receptor indicating that the beta domain in the alphaCGbeta tether was still functional. In contrast, no significant receptor binding of alphaCGbeta/CGbeta was observed indicating a major perturbation in the alpha domain. These results suggest that although dimeric-like determinants are present in both alphaCGbeta/alpha and alphaCGbeta/CGbeta complexes, the receptor binding determinants in the alpha domain of the tether are absent. These results show that generating heterodimeric determinants do not necessarily result in a bioactive molecule. Our data also indicate that the determinants for biological activity are distinct from those associated with intracellular behavior. PMID- 11390410 TI - Succinic acids as potent inhibitors of plasmid-borne IMP-1 metallo-beta lactamase. AB - IMP-1 metallo-beta-lactamase (class B) is a plasmid-borne zinc metalloenzyme that efficiently hydrolyzes beta-lactam antibiotics, including carbapenems, rendering them ineffective. Because IMP-1 has been found in several clinically important carbapenem-resistant pathogens, there is a need for inhibitors of this enzyme that could protect broad spectrum antibiotics such as imipenem from hydrolysis and thus extend their utility. We have identified a series of 2,3-(S,S) disubstituted succinic acids that are potent inhibitors of IMP-1. Determination of high resolution crystal structures and molecular modeling of succinic acid inhibitor complexes with IMP-1 has allowed an understanding of the potency, stereochemistry, and structure-activity relationships of these inhibitors. PMID- 11390411 TI - Small nuclear RNA genes: a model system to study fundamental mechanisms of transcription. PMID- 11390412 TI - COX in a crystal ball: current status and future promise of prostaglandin research. PMID- 11390413 TI - Arachidonic acid as a bioactive molecule. PMID- 11390414 TI - Regulated formation of eicosanoids. PMID- 11390415 TI - Gene therapy for rheumatoid arthritis? PMID- 11390416 TI - Not the usual suspects: the unexpected sources of tissue regeneration. PMID- 11390417 TI - The C-C chemokine receptors CCR4 and CCR8 identify airway T cells of allergen challenged atopic asthmatics. AB - In vitro polarized human Th2 cells preferentially express the chemokine receptors CCR3, CCR4, and CCR8 and migrate to their ligands: eotaxin, monocyte-derived chemokine (MDC), thymus- and activation-regulated chemokine (TARC), and I-309. We have studied the expression of chemokines and chemokine receptors in the airway mucosa of atopic asthmatics. Immunofluorescent analysis of endobronchial biopsies from six asthmatics, taken 24 hours after allergen challenge, demonstrates that virtually all T cells express IL-4 and CCR4. CCR8 is coexpressed with CCR4 on 28% of the T cells, while CCR3 is expressed on eosinophils but not on T cells. Expression of the CCR4-specific ligands MDC and TARC is strongly upregulated on airway epithelial cells upon allergen challenge, suggesting an involvement of this receptor/ligand axis in the regulation of lymphocyte recruitment into the asthmatic bronchi. In contrast to asthma, T cells infiltrating the airways of patients with chronic obstructive pulmonary disease and pulmonary sarcoidosis produce IFN-gamma and express high levels of CXCR3, while lacking CCR4 and CCR8 expression. These data support the role of CCR4, of its ligands MDC and TARC, and of CCR8 in the pathogenesis of allergen-induced late asthmatic responses and suggest that these molecules could be considered as targets for therapeutic intervention. PMID- 11390418 TI - Hepatocyte growth factor ameliorates acute graft-versus-host disease and promotes hematopoietic function. AB - Acute graft-versus-host disease (GVHD) is a major complication of bone marrow transplantation (BMT) and is characterized by hematopoietic dysfunction, immunosuppression, and tissue injury in the skin, liver, and intestinal mucosa. Hepatocyte growth factor (HGF), originally identified and cloned as a potent mitogen for hepatocytes, induces mitogenic and antiapoptotic activity in various epithelial cells and promotes hematopoiesis. Working in a murine model of acute GVHD, we performed repeated transfection of the human HGF cDNA into skeletal muscle and showed that this treatment inhibited apoptosis of intestinal epithelial cells and donor T-cell infiltration into the liver, thereby ameliorating the enteropathy and liver injury caused by acute GVHD. HGF also markedly suppressed IFN-gamma and TNF-alpha expression in the intestine and liver and decreased the serum IL-12. Furthermore, extramedullary hematopoiesis by donor cells was increased, and the survival rate was improved. These results suggest that HGF may be useful for controlling acute GVHD after allogeneic BMT. PMID- 11390419 TI - IL-4 abrogates osteoclastogenesis through STAT6-dependent inhibition of NF kappaB. AB - IL-4, an anti-inflammatory cytokine, inhibits osteoclast differentiation, but the basis of this effect has been unclear. Osteoclastogenesis requires activation of RANK, which exerts its biologic effect via activation of NF-kappaB. NF-kappaB activation is manifested by nuclear translocation and binding to DNA, events secondary to phosphorylation and dissociation of IkappaBalpha. It is shown here that IL-4 reduces NF-kappaB nuclear translocation by inhibiting IkappaB phosphorylation, thus markedly inhibiting NF-kappaB DNA binding activity and blocking osteoclastogenesis entirely. Residual translocation of NF-kappaB in the presence of IL-4, however, suggests that nuclear mechanisms must primarily account for inhibition of NF-kappaB DNA binding and blockade of osteoclastogenesis. To address this issue, this study examined whether IL-4 induced STAT6 transcription factor blocks NF-kappaB transactivation. The results show that excess unlabeled consensus sequence STAT6, but not its mutated form, inhibits NF-kappaB binding. Furthermore, exogenously added STAT6 protein inhibits NF-kappaB/DNA interaction. Further supporting a role for STAT6 in this process are the findings that IL-4 fails to block osteoclastogenesis in STAT6(-/-) mice but that this blockade can be restored with addition of exogenous STAT6. Thus, IL 4 obliterates osteoclast differentiation by antagonizing NF-kappaB activation in a STAT6-dependent manner. PMID- 11390420 TI - LDL receptor-related protein mediates cell-surface clustering and hepatic sequestration of chylomicron remnants in LDLR-deficient mice. AB - It has been proposed that in the liver, chylomicron remnants (lipoproteins carrying dietary lipid) may be sequestered before being internalized by hepatocytes. To study this, chylomicron remnants labeled with a fluorescent dye were perfused into isolated livers of LDL receptor-deficient (LDLR-deficient) mice (Ldlr(-/-)) and examined by confocal microscopy. In contrast to livers from normal mice, there was clustering of the chylomicron remnants on the cell surface in the space of DISSE: These remnant clusters colocalized with clusters of LDLR related protein (LRP) and could be eliminated by low concentrations of receptor associated protein, an inhibitor of LRP. When competed with ligands of heparan sulfate proteoglycans (HSPGs), the remnant clusters still appeared but were fewer in number, although syndecans (membrane HSPGs) colocalized with the remnant clusters. This suggests that the clustering of remnants is not dependent on syndecans but that the syndecans may modify the binding of remnants. These results establish that sequestration is a novel process, the clustering of remnants in the space of DISSE: The clustering involves remnants binding to the LRP, and this may be stabilized by binding with syndecans, eventually followed by endocytosis. PMID- 11390421 TI - Regeneration of ischemic cardiac muscle and vascular endothelium by adult stem cells. AB - Myocyte loss in the ischemically injured mammalian heart often leads to irreversible deficits in cardiac function. To identify a source of stem cells capable of restoring damaged cardiac tissue, we transplanted highly enriched hematopoietic stem cells, the so-called side population (SP) cells, into lethally irradiated mice subsequently rendered ischemic by coronary artery occlusion for 60 minutes followed by reperfusion. The engrafted SP cells (CD34(-)/low, c Kit(+), Sca-1(+)) or their progeny migrated into ischemic cardiac muscle and blood vessels, differentiated to cardiomyocytes and endothelial cells, and contributed to the formation of functional tissue. SP cells were purified from Rosa26 transgenic mice, which express lacZ widely. Donor-derived cardiomyocytes were found primarily in the peri-infarct region at a prevalence of around 0.02% and were identified by expression of lacZ and alpha-actinin, and lack of expression of CD45. Donor-derived endothelial cells were identified by expression of lacZ and Flt-1, an endothelial marker shown to be absent on SP cells. Endothelial engraftment was found at a prevalence of around 3.3%, primarily in small vessels adjacent to the infarct. Our results demonstrate the cardiomyogenic potential of hematopoietic stem cells and suggest a therapeutic strategy that eventually could benefit patients with myocardial infarction. PMID- 11390422 TI - Lack of mitochondrial trifunctional protein in mice causes neonatal hypoglycemia and sudden death. AB - Mitochondrial trifunctional protein (MTP) is a hetero-octamer of four alpha and four beta subunits that catalyzes the final three steps of mitochondrial long chain fatty acid beta-oxidation. Human MTP deficiency causes Reye-like syndrome, cardiomyopathy, or sudden unexpected death. We used gene targeting to generate an MTP alpha subunit null allele and to produce mice that lack MTP alpha and beta subunits. The Mtpa(-/-) fetuses accumulate long chain fatty acid metabolites and have low birth weight compared with the Mtpa(+/-) and Mtpa(+/+) littermates. Mtpa(-/-) mice suffer neonatal hypoglycemia and sudden death 6-36 hours after birth. Analysis of the histopathological changes in the Mtpa(-/-) pups revealed rapid development of hepatic steatosis after birth and, later, significant necrosis and acute degeneration of the cardiac and diaphragmatic myocytes. This mouse model documents that intact mitochondrial long chain fatty acid oxidation is essential for fetal development and for survival after birth. Deficiency of MTP causes fetal growth retardation, neonatal hypoglycemia, and sudden death. PMID- 11390423 TI - Origin of neointimal endothelium and alpha-actin-positive smooth muscle cells in transplant arteriosclerosis. AB - The development of transplant arteriosclerosis (TA) is today's most important problem in clinical organ transplantation. Histologically, TA is characterized by perivascular inflammation and progressive intimal thickening. Current thought on this process of vascular remodeling assumes that neointimal vascular smooth muscle (VSM) cells and endothelium in TA are graft-derived, holding that medial VSM cells proliferate and migrate into the subendothelial space in response to signals from inflammatory cells and damaged graft endothelium. Using MHC class I haplotype-specific immunohistochemical staining and single-cell PCR analyses, we show that the neointimal alpha-actin-positive VSM cells in rat aortic or cardiac allografts are of recipient and not of donor origin. In aortic but not in cardiac allografts, recipient-derived endothelial cells (ECs) replaced donor endothelium. Cyclosporine treatment prevents neointima formation and preserves the vascular media in aortic allografts. Recipient-derived ECs do not replace graft endothelium after cyclosporine treatment. We propose that, although it progresses beyond the needs of functional repair, TA reflects the activity of a normal healing process that restores vascular wall function following allograft-induced immunological injury. PMID- 11390425 TI - T-cell involvement in drug-induced acute generalized exanthematous pustulosis. AB - Acute generalized exanthematous pustulosis (AGEP) is an uncommon eruption most often provoked by drugs, by acute infections with enteroviruses, or by mercury. It is characterized by acute, extensive formation of nonfollicular sterile pustules on erythematous background, fever, and peripheral blood leukocytosis. We present clinical and immunological data on four patients with this disease, which is caused by different drugs. An involvement of T cells could be implied by positive skin patch tests and lymphocyte transformation tests. Immunohistochemistry revealed a massive cell infiltrate consisting of neutrophils in pustules and T cells in the dermis and epidermis. Expression of the potent neutrophil-attracting chemokine IL-8 was elevated in keratinocytes and infiltrating mononuclear cells. Drug-specific T cells were generated from the blood and skin of three patients, and phenotypic characterization showed a heterogeneous distribution of CD4/CD8 phenotype and of T-cell receptor Vbeta expression. Analysis of cytokine/chemokine profiles revealed that IL-8 is produced significantly more by drug-specific T cells from patients with AGEP compared with drug-specific T cells from patients that had non-AGEP exanthemas. In conclusion, our data demonstrate the involvement of drug-specific T cells in the pathomechanism of this rather rare and peculiar form of drug allergy. In addition, they indicate that even in some neutrophil-rich inflammatory responses specific T cells are engaged and might orchestrate the immune reaction. PMID- 11390424 TI - Statin-induced inhibition of the Rho-signaling pathway activates PPARalpha and induces HDL apoA-I. AB - Statins are inhibitors of the rate-limiting enzyme in cholesterol synthesis, 3 hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. In addition to reducing LDL cholesterol, statin treatment increases the levels of the antiatherogenic HDL and its major apolipoprotein apoA-I. Here, we investigated the molecular mechanisms of apoA-I regulation by statins. Treatment with statins increased apoA I mRNA levels in human HepG2 hepatoma cells, and this effect was reversed by the addition of mevalonate, implicating HMG-CoA reductase as the relevant target of these drugs. Pretreatment with Actinomycin D abolished the increase of apoA-I mRNA, indicating that statins act at the transcriptional level. Indeed, statins increased the human apoA-I promoter activity in transfected cells, and we have identified a statin response element that coincides with a PPARalpha response element known to confer fibrate responsiveness to this gene. The statin effect could be abolished not only by mevalonate, but also by geranylgeranyl pyrophosphate, whereas inhibition of geranylgeranyl transferase activity or treatment with an inhibitor of the Rho GTP-binding protein family increased PPARalpha activity. Using dominant negative forms of these proteins, we found that Rho A itself mediates this response. Because cotreatment with statins and fibrates activated PPARalpha in a synergistic manner, these observations provide a molecular basis for combination treatment with statins and fibrates in coronary heart disease. PMID- 11390426 TI - The protease-activated receptor-2 agonist induces gastric mucus secretion and mucosal cytoprotection. AB - Protease-activated receptor-2 (PAR-2), a receptor activated by trypsin/tryptase, modulates smooth muscle tone and exocrine secretion in the salivary glands and pancreas. Given that PAR-2 is expressed throughout the gastrointestinal tract, we investigated effects of PAR-2 agonists on mucus secretion and gastric mucosal injury in the rat. PAR-2-activating peptides triggered secretion of mucus in the stomach, but not in the duodenum. This mucus secretion was abolished by pretreatment with capsaicin, which stimulates and ablates specific sensory neurons, but it was resistant to cyclo-oxygenase inhibition. In contrast, capsaicin treatment failed to block PAR-2-mediated secretion from the salivary glands. Intravenous calcitonin gene-related peptide (CGRP) and neurokinin A markedly elicited gastric mucus secretion, as did substance P to a lesser extent. Specific antagonists of the CGRP1 and NK2, but not the NK1, receptors inhibited PAR-2-mediated mucus secretion. Pretreatment with the PAR-2 agonist strongly prevented gastric injury caused by HCl-ethanol or indomethacin. Thus, PAR-2 activation triggers the cytoprotective secretion of gastric mucus by stimulating the release of CGRP and tachykinins from sensory neurons. In contrast, the PAR-2 mediated salivary exocrine secretion appears to be independent of capsaicin sensitive sensory neurons. PMID- 11390427 TI - TNF-alpha downregulates murine hepatic growth hormone receptor expression by inhibiting Sp1 and Sp3 binding. AB - Children with chronic inflammatory diseases experience growth failure and wasting. This may be due to growth hormone resistance caused by cytokine-induced suppression of growth hormone receptor (GHR) gene expression. However, the factors governing inflammatory regulation of GHR are not known. We have reported that Sp1 and Sp3 regulate hepatic GHR expression. We hypothesized that TNF-alpha suppresses GHR expression by inhibiting Sp1/Sp3 transactivators. LPS administration significantly reduced murine hepatic GHR expression, as well as Sp1 and Sp3 binding to GHR promoter cis elements. TNF-alpha was integral to this response, as LPS did not affect hepatic Sp1/Sp3 binding or GHR expression in TNF receptor 1-deficient mice. TNF-alpha treatment of BNL CL.2 mouse liver cells reduced Sp1 and Sp3 binding to a GHR promoter cis element and downregulated activity of a GHR promoter-driven luciferase reporter. Combined mutations within adjacent Sp elements eliminated GHR promoter suppression by TNF-alpha without affecting overall nuclear levels of Sp1 or Sp3 proteins. These studies demonstrate that murine GHR transcription is downregulated by LPS, primarily via TNF-alpha-dependent signaling. Evidence suggests that inhibition of Sp transactivator binding is involved. Further investigation of these mechanisms may identify novel strategies for preventing inflammatory suppression of growth. PMID- 11390428 TI - Modulation of T-cell responses to alloantigens by TR6/DcR3. AB - TR6 (DcR3) is a new member of the TNF receptor (TNFR) family that lacks a transmembrane domain in its sequence, indicating that it is a secreted molecule. TR6 can bind to FasL and prevent FasL-induced apoptosis; it can also associate with LIGHT, another TNF family member. The role of TR6 in immune responses was investigated in this study. According to flow cytometry, recombinant human TR6-Fc binds to human LIGHT expressed on 293 cells or on activated human T cells and competes with the LIGHT receptor TR2 for the binding to LIGHT on these cells. Human TR6 could cross-react with mouse LIGHT in immunoprecipitation. TR6-Fc also downregulates cytotoxic T lymphocyte activity in vitro and graft-versus-host responses in mice. Moreover, TR6-Fc modulates lymphokine production by alloantigen-stimulated mouse T cells. TR6-Fc ameliorated rejection response to mouse heart allograft. These results indicate that TR6 can dampen T-cell responses to alloantigens. Such regulatory effects of TR6 probably occur via interference with interaction between pairs of related TNF and TNFR family members, LIGHT/TR2 being one of the possible candidate pairs. PMID- 11390429 TI - Relaxin is essential for renal vasodilation during pregnancy in conscious rats. AB - Marked vasodilation in the kidney and other nonreproductive organs is one of the earliest maternal adaptations to occur during pregnancy. Despite the recognition of this extraordinary physiology for over four decades, the gestational hormone responsible has remained elusive. Here we demonstrate a key role for relaxin, a member of the IGF family that is secreted by the corpus luteum in humans and rodents. Using a gravid rodent model, we employ two approaches to eliminate relaxin or its biological activity from the circulation: ovariectomy and administration of neutralizing antibodies. Both abrogate the gestational elevation in renal perfusion and glomerular filtration, as well as preventing the reduction in myogenic reactivity of isolated, small renal arteries. Osmoregulatory changes, another pregnancy adaptation, are also abolished. Our results indicate that relaxin mediates the renal vasodilatory responses to pregnancy and thus may be important for maternal and fetal health. They also raise the likelihood of a role for relaxin in other cardiovascular changes of pregnancy, and they suggest that, like estrogen, relaxin should be considered a regulator of cardiovascular function. PMID- 11390431 TI - T cell recognition of the dominant I-A(k)-restricted hen egg lysozyme epitope: critical role for asparagine deamidation. AB - Type-B T cells raised against the immunodominant peptide in hen egg lysozyme (HEL(48-62)) do not respond to whole lysozyme, and this has been thought to indicate that peptide can bind to l-A(k) in different conformations. Here we demonstrate that such T cells recognize a deamidated form of the HEL peptide and not the native peptide. The sequence of the HEL epitope facilitates rapid and spontaneous deamidation when present as a free peptide or within a flexible domain. However, this deamidated epitope is not created within intact lysozyme, most likely because it resides in a highly structured part of the protein. These findings argue against the existence of multiple conformations of the same peptide-MHC complex and have important implications for the design of peptide based vaccines. Furthermore, as the type-B T cells are known to selectively evade induction of tolerance when HEL is expressed as a transgene, these results suggest that recognition of posttranslationally modified self-antigen may play a role in autoimmunity. PMID- 11390430 TI - Essential role of lymph nodes in contact hypersensitivity revealed in lymphotoxin alpha-deficient mice. AB - Lymph nodes (LNs) are important sentinal organs, populated by circulating lymphocytes and antigen-bearing cells exiting the tissue beds. Although cellular and humoral immune responses are induced in LNs by antigenic challenge, it is not known if LNs are essential for acquired immunity. We examined immune responses in mice that lack LNs due to genetic deletion of lymphotoxin ligands or in utero blockade of membrane lymphotoxin. We report that LNs are absolutely required for generating contact hypersensitivity, a T cell-dependent cellular immune response induced by epicutaneous hapten. We show that the homing of epidermal Langerhans cells in response to hapten application is specifically directed to LNs, providing a cellular basis for this unique LN function. In contrast, the spleen cannot mediate contact hypersensitivity because antigen-bearing epidermal Langerhans cells do not access splenic white pulp. Finally, we formally demonstrate that LNs provide a unique environment essential for generating this acquired immune response by reversing the LN defect in lymphotoxin-alpha(-/)- mice, thereby restoring the capacity for contact hypersensitivity. PMID- 11390432 TI - Treatment of allergic airway inflammation and hyperresponsiveness by antisense induced local blockade of GATA-3 expression. AB - Recent studies in transgenic mice have revealed that expression of a dominant negative form of the transcription factor GATA-3 in T cells can prevent T helper cell type 2 (Th2)-mediated allergic airway inflammation in mice. However, it remains unclear whether GATA-3 plays a role in the effector phase of allergic airway inflammation and whether antagonizing the expression and/or function of GATA-3 can be used for the therapy of allergic airway inflammation and hyperresponsiveness. Here, we analyzed the effects of locally antagonizing GATA-3 function in a murine model of asthma. We could suppress GATA-3 expression in interleukin (IL)-4-producing T cells in vitro and in vivo by an antisense phosphorothioate oligonucleotide overlapping the translation start site of GATA 3, whereas nonsense control oligonucleotides were virtually inactive. In a murine model of asthma associated with allergic pulmonary inflammation and hyperresponsiveness in ovalbumin (OVA)-sensitized mice, local intranasal administration of fluorescein isothiocyanate-labeled GATA-3 antisense oligonucleotides led to DNA uptake in lung cells associated with a reduction of intracellular GATA-3 expression. Such intrapulmonary blockade of GATA-3 expression caused an abrogation of signs of lung inflammation including infiltration of eosinophils and Th2 cytokine production. Furthermore, treatment with antisense but not nonsense oligonucleotides induced a significant reduction of airway hyperresponsiveness in OVA-sensitized mice to levels comparable to saline-treated control mice, as assessed by both enhanced pause (PenH) responses and pulmonary resistance determined by body plethysmography. These data indicate a critical role for GATA-3 in the effector phase of a murine asthma model and suggest that local delivery of GATA-3 antisense oligonucleotides may be a novel approach for the treatment of airway hyperresponsiveness such as in asthma. This approach has the potential advantage of suppressing the expression of various proinflammatory Th2 cytokines simultaneously rather than suppressing the activity of a single cytokine. PMID- 11390433 TI - Adjuvant immunotherapy is dependent on inducible nitric oxide synthase. AB - Rodents immunized with complete Freund's adjuvant (CFA) are resistant to subsequent attempts to induce autoimmune disease, while animals immunized with incomplete Freund's adjuvant (IFA) remain susceptible. Mycobacterial extracts can stimulate inducible nitric oxide synthase (NOS2) gene transcription. Robust expression of NOS2 has been linked to suppression of T cell proliferation and alterations in immune responses. Our studies investigated the hypothesis that the immunoprotective effect of CFA before immunization requires functional NOS2. NOS2 gene expression is chronically elevated in lymph nodes and spleens of CFA immunized mice. Maximal expression of NOS2 after CFA immunization requires the presence of functional type I tumor necrosis factor alpha receptor (TNFR1) and interferon gamma. Groups of nontreated and CFA-preimmunized male C57BL/6J or C57BL/6NOS2(-/)- mice were immunized with myelin oligodendrocyte glycoprotein (MOG) peptide 35-55 in CFA to induce experimental allergic encephalomyelitis (EAE). Wild-type C57BL/6J mice were protected from the development of symptoms of EAE, while the NOS2(-/)- mice failed to be protected. NOS2-dependent effects of CFA included an augmentation of the MOG-specific IgG1 response, a decrease in interleukin 6 production by MOG-reactive lymphocytes, and a marked decrease in mononuclear cell infiltrates in the central nervous system. These studies support the hypothesis that CFA immunization modulates immune responses through a nitric oxide-dependent mechanism. PMID- 11390434 TI - The transmembrane adaptor protein TRIM regulates T cell receptor (TCR) expression and TCR-mediated signaling via an association with the TCR zeta chain. AB - T cell receptor (TCR)-interacting molecule (TRIM) is a recently identified transmembrane adaptor protein, which is exclusively expressed in T cells. Here we demonstrate that in mature T cells, TRIM preferentially interacts with the TCR via the TCR-zeta chains and to a lesser extent via the CD3-straightepsilon/gamma heterodimer. Transient or stable overexpression of TRIM in Jurkat T cells results in enhancement of TCR expression on the cell surface and elevated induction of Ca(2+) mobilization after T cell activation. TRIM-mediated upregulation of TCR expression results from inhibition of spontaneous TCR internalization and stabilization of TCR complexes on the cell surface. Collectively, our data identify TRIM as a novel integral component of the TCR complex and suggest that one function of TRIM might be to modulate the strength of signals transduced through the TCR through regulation of TCR expression on the cell surface. PMID- 11390435 TI - Identification and functional characterization of human CD4(+)CD25(+) T cells with regulatory properties isolated from peripheral blood. AB - A subpopulation of peripheral human CD4(+)CD25(+) T cells that expresses CD45RO, histocompatibility leukocyte antigen DR, and intracellular cytotoxic T lymphocyte associated antigen (CTLA) 4 does not expand after stimulation and markedly suppresses the expansion of conventional T cells in a contact-dependent manner. After activation, CD4(+)CD25(+) T cells express CTLA-4 on the surface detectable for several weeks. These cells show a G1/G0 cell cycle arrest and no production of interleukin (IL)-2, IL-4, or interferon (IFN)-gamma on either protein or mRNA levels. The anergic state of CD4(+)CD25(+) T cells is not reversible by the addition of anti-CD28, anti-CTLA-4, anti-transforming growth factor beta, or anti IL-10 antibody. However, the refractory state of CD4(+)CD25(+) T cells was partially reversible by the addition of IL-2 or IL-4. These data demonstrate that human blood contains a resident T cell population with potent regulatory properties. PMID- 11390436 TI - Human cd25(+)cd4(+) t regulatory cells suppress naive and memory T cell proliferation and can be expanded in vitro without loss of function. AB - Active suppression by T regulatory (Tr) cells plays an important role in the downregulation of T cell responses to foreign and self-antigens. Mouse CD4(+) Tr cells that express CD25 possess remarkable suppressive activity in vitro and in autoimmune disease models in vivo. Thus far, the existence of a similar subset of CD25(+)CD4(+) Tr cells in humans has not been reported. Here we show that human CD25(+)CD4(+) Tr cells isolated from peripheral blood failed to proliferate and displayed reduced expression of CD40 ligand (CD40L), in response to T cell receptor-mediated polyclonal activation, but strongly upregulated cytotoxic T lymphocyte-associated antigen (CTLA)-4. Human CD25(+)CD4(+) Tr cells also did not proliferate in response to allogeneic antigen-presenting cells, but they produced interleukin (IL)-10, transforming growth factor (TGF)-beta, low levels of interferon (IFN)-gamma, and no IL-4 or IL-2. Importantly, CD25(+)CD4(+) Tr cells strongly inhibited the proliferative responses of both naive and memory CD4(+) T cells to alloantigens, but neither IL-10, TGF-beta, nor CTLA-4 seemed to be directly required for their suppressive effects. CD25(+)CD4(+) Tr cells could be expanded in vitro in the presence of IL-2 and allogeneic feeder cells and maintained their suppressive capacities. These findings that CD25(+)CD4(+) Tr cells with immunosuppressive effects can be isolated from peripheral blood and expanded in vitro without loss of function represent a major advance towards the therapeutic use of these cells in T cell-mediated diseases. PMID- 11390437 TI - Ex vivo isolation and characterization of CD4(+)CD25(+) T cells with regulatory properties from human blood. AB - It has been known for years that rodents harbor a unique population of CD4(+)CD25(+) "professional" regulatory/suppressor T cells that is crucial for the prevention of spontaneous autoimmune diseases. Here we demonstrate that CD4(+)CD25(+)CD45RO(+) T cells (mean 6% of CD4(+) T cells) are present in the blood of adult healthy volunteers. In contrast to previous reports, these CD4(+)CD25(+) T cells do not constitute conventional memory cells but rather regulatory cells exhibiting properties identical to their rodent counterparts. Cytotoxic T lymphocyte-associated antigen (CTLA)-4 (CD152), for example, which is essential for the in vivo suppressive activity of CD4(+)CD25(+) T cells, was constitutively expressed, and remained strongly upregulated after stimulation. The cells were nonproliferative to stimulation via their T cell receptor for antigen, but the anergic state was partially reversed by interleukin (IL)-2 and IL-15. Upon stimulation with allogeneic (but not syngeneic) mature dendritic cells or platebound anti-CD3 plus anti-CD28 the CD4(+)CD25(+) T cells released IL 10, and in coculture experiments suppressed the activation and proliferation of CD4(+) and CD8(+) T cells. Suppression proved IL-10 independent, yet contact dependent as in the mouse. The identification of regulatory CD4(+)CD25(+) T cells has important implications for the study of tolerance in man, notably in the context of autoimmunity, transplantation, and cancer. PMID- 11390438 TI - CD4(+)CD25(+) immune regulatory cells are required for induction of tolerance to alloantigen via costimulatory blockade. AB - Immune regulatory CD4(+)CD25(+) cells play a vital role in the induction and maintenance of self-tolerance and are essential for T cell homeostasis and the prevention of autoimmunity. Induction of tolerance to allogeneic donor grafts is a clinically desirable goal in bone marrow and solid organ transplantation. To determine whether CD4(+)CD25(+) cells regulate T cell responses to alloantigen and are critical for tolerance induction, murine CD4(+) T cells were tolerized to alloantigen via ex vivo CD40 ligand (CD40L)/CD40 or CD28/cytotoxic T lymphocyte associated antigen 4/B7 blockade resulting in secondary mixed leukocyte reaction hyporesponsiveness and tolerance to alloantigen in vivo. CD4(+)CD25(+) T cells were found to be potent regulators of alloresponses. Depletion of CD4(+)CD25(+) T cells from the CD4(+) responder population completely abrogated ex vivo tolerance induction to alloantigen as measured by intact responses to alloantigen restimulation in vitro and in vivo. Addback of CD4(+)CD25(+) T cells to CD4(+)CD25(-) cultures restored tolerance induction. These data are the first to indicate that CD4(+)CD25(+) cells are essential for the induction of tolerance to alloantigen and have important implications for tolerance-inducing strategies targeted at T cell costimulatory pathways. PMID- 11390439 TI - Immunoproteasomes shape immunodominance hierarchies of antiviral CD8(+) T cells at the levels of T cell repertoire and presentation of viral antigens. AB - Vertebrates express three cytokine-inducible proteasome subunits that are incorporated in the place of their constitutively synthesized counterparts. There is increasing evidence that the set of peptides generated by proteasomes containing these subunits (immunoproteasomes) differs from that produced by standard proteasomes. In this study, we use mice lacking one of the immunoproteasome subunits (LMP2) to show that immunoproteasomes play an important role in establishing the immunodominance hierarchy of CD8(+) T cells (T(CD8+)) responding to seven defined determinants in influenza virus. In LMP2(-/)- mice, responses to the two most dominant determinants drop precipitously, whereas responses to two subdominant determinants are greatly enhanced. Adoptive transfer experiments with naive normal and transgenic T(CD8+) reveal that the reduced immunogenicity of one determinant (PA(224-233)) can be attributed to decreased generation by antigen presenting cells (APCs), whereas the other determinant (NP(366-374)) is less immunogenic due to alterations in the T(CD8+) repertoire, and not, as reported previously, to the decreased capacity of LMP2(-/)- APCs to generate the determinant. The enhanced response to one of the subdominant determinants (PB1F2(62-70)) correlates with increased generation by LMP2(-)(/)- virus-infected cells. These findings indicate that in addition to their effects on the presentation of foreign antigens, immunoproteasomes influence T(CD8+) responses by modifying the repertoire of responding T(CD8+). PMID- 11390440 TI - Reversal of spontaneous autoimmune insulitis in nonobese diabetic mice by soluble lymphotoxin receptor. AB - One striking feature of spontaneous autoimmune diabetes is the prototypic formation of lymphoid follicular structures within the pancreas. Lymphotoxin (LT) has been shown to play an important role in the formation of lymphoid follicles in the spleen. To explore the potential role of LT-mediated microenvironment in the pathogenesis of insulin-dependent diabetes mellitus (IDDM), an LTbeta receptor-immunoglobulin fusion protein (LTbetaR-Ig) was administered to nonobese diabetic mice. Early treatment with LTbetaR-Ig prevented insulitis and IDDM, suggesting that LT plays a critical role in the insulitis development. LTbetaR-Ig treatment at a late stage of the disease also dramatically reversed insulitis and prevented diabetes. Moreover, LTbetaR-Ig treatment prevented the development of IDDM by diabetogenic T cells in an adoptive transfer model. Thus, LTbetaR-Ig can disassemble the well established lymphoid microenvironment in the islets, which is required for the development and progression of IDDM. PMID- 11390441 TI - A critical role for lymphotoxin-beta receptor in the development of diabetes in nonobese diabetic mice. AB - To assess the role of lymphotoxin-beta receptor (LTbetaR) in diabetes pathogenesis, we expressed an LTbetaR-Fc fusion protein in nonobese diabetic (NOD) mice. The fusion protein was expressed in the embryo, reached high levels for the first 2 wk after birth, and then declined progressively with age. High expression of LTbetaR-Fc blocked diabetes development but not insulitis. After the decline in chimeric protein concentration, mice became diabetic with kinetics similar to the controls. Early expression of fusion protein resulted in disrupted splenic architecture. However, primary follicles and follicular dendritic cells, but not marginal zones, developed in aged mice. Hence, LTbetaR signaling is required for diabetes development and regulates follicular and marginal zone structures via qualitatively or quantitatively distinct mechanisms. PMID- 11390442 TI - Certified professionals: CD4(+)CD25(+) suppressor T cells. PMID- 11390443 TI - Cutting edge: lipid raft integrity affects the efficiency of MHC class I tetramer binding and cell surface TCR arrangement on CD8+ T cells. AB - Physically distinct cholesterol/sphingolipid-rich plasma membrane microdomains, so-called lipid rafts, have been recognized to play an important regulatory role in various cellular processes, from membrane trafficking to signal transduction, in a number of cell types. We report here that the ability of TCR on activated, functional CD8+ T lymphocytes to efficiently bind MHC class I tetramer complexes is dependent on the integrity of lipid rafts on the T lymphocyte membrane. We further provide evidence that TCR interact (associate) with lipid raft elements on the T cell surface before receptor engagement and that the topological arrangement of TCR on the cell surface is likewise influenced by lipid raft integrity. PMID- 11390445 TI - Neutrophil Fc gamma RI as target for immunotherapy of invasive candidiasis. AB - Invasive candidiasis represents a life-threatening disease for immunocompromised patients. This study focused on new immunotherapeutic approaches for systemic Candida albicans infections in a human FcgammaRI-transgenic mouse model. FcgammaRI (CD64) is a potent immunoactivating receptor on phagocytic and dendritic cells. In vivo targeting of C. albicans toward neutrophil-FcgammaRI by bispecific Abs and G-CSF effectively protected FcgammaRI-transgenic mice from lethal candidiasis. Nontransgenic mice were not protected, and treatment with bispecific Ab or G-CSF alone did not reduce mortality. Furthermore, infected FcgammaRI-transgenic mice developed high titers of anti-C. albicans IgG, and survival was extended on secondary infection without further treatment. These findings document the capacity of FcgammaRI to initiate potent anti-C. albicans immunity and support the development of FcgammaRI-directed immunotherapy of invasive fungal disease. PMID- 11390444 TI - Cutting edge: IL-18-transgenic mice: in vivo evidence of a broad role for IL-18 in modulating immune function. AB - IL-18 has been shown to be a strong cofactor for Th1 T cell development. However, we previously demonstrated that when IL-18 was combined with IL-2, there was a synergistic induction of a Th2 cytokine, IL-13, in both T and NK cells. More recently, we and other groups have reported that IL-18 can potentially induce IgE, IgG1, and Th2 cytokine production in murine experimental models. Here, we report on the generation of IL-18-transgenic (Tg) mice in which mature mouse IL 18 cDNA was expressed. CD8+CD44high T cells and macrophages were increased, but B cells were decreased in these mice while serum IgE, IgG1, IL-4, and IFN-gamma levels were significantly increased. Splenic T cells in IL-18 Tg mice produced higher levels of IFN-gamma, IL-4, IL-5, and IL-13 than control wild-type mice. Thus, aberrant expression of IL-18 in vivo results in the increased production of both Th1 and Th2 cytokines. PMID- 11390446 TI - A dual role for TNF-alpha in type 1 diabetes: islet-specific expression abrogates the ongoing autoimmune process when induced late but not early during pathogenesis. AB - We report here that islet-specific expression of TNF-alpha can play a dual role in autoimmune diabetes, depending on its precise timing in relation to the ongoing autoimmune process. In a transgenic model (rat insulin promoter lymphocytic choriomeningitis virus) of virally induced diabetes, TNF-alpha enhanced disease incidence when induced through an islet-specific tetracycline dependent promoter system early during pathogenesis. Blockade of TNF-alpha during this phase prevented diabetes completely, suggesting its pathogenetic importance early in disease development. In contrast, TNF-alpha expression abrogated the autoimmune process when induced late, which was associated with a reduction of autoreactive CD8 lymphocytes in islets and their lytic activities. Thus, the fine tuned kinetics of an autoreactive process undergo distinct stages that respond in a differential way to the presence of TNF-alpha. This observation has importance for understanding the complex role of inflammatory cytokines in autoimmunity. PMID- 11390447 TI - Infection of human macrophages and dendritic cells with Mycobacterium tuberculosis induces a differential cytokine gene expression that modulates T cell response. AB - Macrophages and dendritic cells (DC) play an essential role in the initiation and maintenance of immune response to pathogens. To analyze early interactions between Mycobacterium tuberculosis (Mtb) and immune cells, human peripheral blood monocyte-derived macrophages (MDM) and monocyte-derived dendritic cells (MDDC) were infected with Mtb. Both cells were found to internalize the mycobacteria, resulting in the activation of MDM and maturation of MDDC as reflected by enhanced expression of several surface Ags. After Mtb infection, the proinflammatory cytokines TNF-alpha, IL-1, and IL-6 were secreted mainly by MDM. As regards the production of IFN-gamma-inducing cytokines, IL-12 and IFN-alpha, was seen almost exclusively from infected MDDC, while IL-18 was secreted preferentially by macrophages. Moreover, Mtb-infected MDM also produce the immunosuppressive cytokine IL-10. Because IL-10 is a potent inhibitor of IL-12 synthesis from activated human mononuclear cells, we assessed the inhibitory potential of this cytokine using soluble IL-10R. Neutralization of IL-10 restored IL-12 secretion from Mtb-infected MDM. In line with these findings, supernatants from Mtb-infected MDDC induced IFN-gamma production by T cells and enhanced IL 18R expression, whereas supernatants from MDM failed to do that. Neutralization of IFN-alpha, IL-12, and IL-18 activity in Mtb-infected MDDC supernatants by specific Abs suggested that IL-12 and, to a lesser extent, IFN-alpha and IL-18 play a significant role in enhancing IFN-gamma synthesis by T cells. During Mtb infection, macrophages and DC may have different roles: macrophages secrete proinflammatory cytokines and induce granulomatous inflammatory response, whereas DC are primarily involved in inducing antimycobacterial T cell immune response. PMID- 11390448 TI - Liver-derived DEC205+B220+CD19- dendritic cells regulate T cell responses. AB - Leukocytes resident in the liver may play a role in immune responses. We describe a cell population propagated from mouse liver nonparenchymal cells in IL-3 and anti-CD40 mAb that exhibits a distinct surface immunophenotype and function in directing differentiation of naive allogeneic T cells. After culture, such cells are DEC-205(bright)B220+CD11c-CD19-, and negative for T (CD3, CD4, CD8alpha), NK (NK 1.1) cell markers, and myeloid Ags (CD11b, CD13, CD14). These liver-derived DEC205+B220+ CD19- cells have a morphology and migratory capacity similar to dendritic cells. Interestingly, they possess Ig gene rearrangements, but lack Ig molecule expression on the cell surface. They induce low thymidine uptake of allogeneic T cells in MLR due to extensive apoptosis of activated T cells. T cell proliferation is restored by addition of the common caspase inhibitor peptide, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone (zVAD-fmk). T cells stimulated by liver-derived DEC205+B220+D19- cells release both IL-10 and IFN-gamma, small amounts of TGF-beta, and no IL-2 or IL-4, a cytokine profile resembling T regulatory type 1 cells. Expression of IL-10 and IFN-gamma, but not bioactive IL 12 in liver DEC205+B220+CD19- cells was demonstrated by RNase protection assay. In vivo administration of liver DEC205+B220+CD19- cells significantly prolonged the survival of vascularized cardiac allografts in an alloantigen-specific manner. PMID- 11390449 TI - Aspirin inhibits in vitro maturation and in vivo immunostimulatory function of murine myeloid dendritic cells. AB - Aspirin is the most commonly used analgesic and antiinflammatory agent. In this study, at physiological concentrations, it profoundly inhibited CD40, CD80, CD86, and MHC class II expression on murine, GM-CSF + IL-4 stimulated, bone marrow derived myeloid dendritic cells (DC). CD11c and MHC class I expression were unaffected. The inhibitory action was dose dependent and was evident at concentrations higher than those necessary to inhibit PG synthesis. Experiments with indomethacin revealed that the effects of aspirin on DC maturation were cyclooxygenase independent. Nuclear extracts of purified, aspirin-treated DC revealed a decreased NF-kappaB DNA-binding activity, whereas Ab supershift analysis indicated that aspirin targeted primarily NF-kappaB p50. Unexpectedly, aspirin promoted the generation of CD11c+ DC, due to apparent suppression of granulocyte development. The morphological and ultrastructural appearance of aspirin-treated cells was consistent with immaturity. Aspirin-treated DC were highly efficient at Ag capture, via both mannose receptor-mediated endocytosis and macropinocytosis. By contrast, they were poor stimulators of naive allogeneic T cell proliferation and induced lower levels of IL-2 in responding T cells. They also exhibited impaired IL-12 expression and did not produce IL-10 after LPS stimulation. Assessment of the in vivo function of aspirin-treated DC, pulsed with the hapten trinitrobenzenesulfonic acid, revealed an inability to induce normal cell-mediated contact hypersensitivity, despite the ability of the cells to migrate to T cell areas of draining lymphoid tissue. These data provide new insight into the immunopharmacology of aspirin and suggest a novel approach to the manipulation of DC for therapeutic application. PMID- 11390450 TI - TAP-independent presentation of CTL epitopes by Trojan antigens. AB - The majority of CTL epitopes are derived from intracellular proteins that are degraded in the cytoplasm by proteasomes into peptides that are transported into the endoplasmic reticulum by the TAP complex. These peptides can be further processed into the optimal size (8-10 residues) for binding with nascent MHC class I molecules, generating complexes that are exported to the cell surface. Proteins or peptides containing CTL epitopes can be introduced into the cytoplasm of APCs by linking them to membrane-translocating Trojan carriers allowing their incorporation into the MHC class I Ag-processing pathway. The present findings suggest that these "Trojan" Ags can be transported into the endoplasmic reticulum in a TAP-independent way where they are processed and trimmed into CTL epitopes. Furthermore, processing of Trojan Ags can also occur in the trans-Golgi compartment, with the participation of the endopeptidase furin and possibly with the additional participation of a carboxypeptidase. We believe that these findings will be of value for the design of CTL-inducing vaccines for the treatment or prevention of infectious and malignant diseases. PMID- 11390451 TI - IFN-gamma alters the pathology of graft rejection: protection from early necrosis. AB - We studied the effect of host IFN-gamma on the pathology of acute rejection of vascularized mouse heart and kidney allografts. Organs from CBA donors (H-2k) were transplanted into BALB/c (H-2d) hosts with wild-type (WT) or disrupted (GKO, BALB/c mice with disrupted IFN-gamma genes) IFN-gamma genes. In WT hosts, rejecting hearts and kidneys showed mononuclear cell infiltration, intense induction of donor MHC products, but little parenchymal necrosis at day 7. Rejecting allografts in GKO recipients showed infiltrate but little or no induction of donor MHC and developed extensive necrosis despite patent large vessels. The necrosis was immunologically mediated, since it developed during rejection, was absent in isografts, and was prevented by immunosuppressing the recipient with cyclosporine or mycophenolate mofetil. Rejecting kidneys in GKO hosts showed increased mRNA for heme oxygenase 1, and decreased mRNA for NO synthase 2 and monokine inducible by IFN-gamma (MIG). The mRNA levels for CTL genes (perforin, granzyme B, and Fas ligand) were similar in rejecting kidneys in WT and GKO hosts, and the host Ab responses were similar. The administration of recombinant IFN-gamma to GKO hosts reduced but did not fully prevent the effects of IFN-gamma deficiency: MHC was induced, but the prevention of necrosis and induction of MIG were incomplete compared with WT hosts. Thus, IFN-gamma has unique effects in vascularized allografts, including induction of MHC and MIG, and protection against parenchymal necrosis, probably at the level of the microcirculation. This is probably a local action of IFN-gamma produced in large quantities in the allograft. PMID- 11390453 TI - Cloning and characterization of IL-22 binding protein, a natural antagonist of IL 10-related T cell-derived inducible factor/IL-22. AB - The class II cytokine receptor family includes the receptors for IFN-alphabeta, IFN-gamma, IL-10, and IL-10-related T cell-derived inducible factor/IL-22. By screening genomic DNA databases, we identified a gene encoding a protein of 231 aa, showing 33 and 34% amino acid identity with the extracellular domains of the IL-22 receptor and of the IL-20R/cytokine receptor family 2-8, respectively, but lacking the transmembrane and cytoplasmic domains. A lower but significant sequence identity was found with other members of this family such as the IL-10R (29%), cytokine receptor family 2-4/IL-10Rbeta (30%), tissue factor (26%), and the four IFN receptor chains (23-25%). This gene is located on chromosome 6q24, at 35 kb from the IFNGR1 gene, and is expressed in various tissues with maximal expression in breast, lungs, and colon. The recombinant protein was found to bind IL-10-related T cell-derived inducible factor/IL-22, and to inhibit the activity of this cytokine on hepatocytes and intestinal epithelial cells. We propose to name this natural cytokine antagonist IL-22BP for IL-22 binding protein. PMID- 11390452 TI - Suppressors of cytokine signaling (SOCS)-1 and SOCS-3 are induced by CpG-DNA and modulate cytokine responses in APCs. AB - During infection, the functional status of the innate immune system is tightly regulated. Although signals resulting in activation have been well characterized, counterregulative mechanisms are poorly understood. Suppressor of cytokine signaling (SOCS) proteins have been characterized as cytokine-inducible negative regulators of Janus kinase/STAT signaling in cells of hemopoietic origin. To analyze whether SOCS proteins could also be induced by pathogen-derived stimuli, we investigated the induction of SOCS-1 and SOCS-3 after triggering of macrophage cell lines, bone marrow-derived dendritic cells, and peritoneal macrophages with CpG-DNA. In this study, we show that CpG-DNA, but not GpC-DNA, induces expression of mRNA for SOCS-1 and SOCS-3 in vitro and in vivo. SOCS mRNA expression could be blocked by chloroquine and was independent of protein synthesis. Inhibitors of the mitogen-activated protein kinase pathway triggered by CpG-DNA were able to impede induction of SOCS mRNA. CpG-DNA triggered synthesis of SOCS proteins that could be detected by Western blotting. SOCS proteins were functional because they inhibited IFN-gamma as well as IL-6- and GM-CSF-induced phosphorylation of STAT proteins. Furthermore, IFN-gamma-induced up-regulation of MHC class II molecules was also prevented. The same effects could be achieved by overexpression of SOCS 1. Hence, the results indicate a substantial cross-talk between signal pathways within cells. They provide evidence for regulative mechanisms of Janus kinase/STAT signaling after triggering Toll-like receptor signal pathways. PMID- 11390454 TI - Identification, cloning, and characterization of a novel soluble receptor that binds IL-22 and neutralizes its activity. AB - With the use of a partial sequence of the human genome, we identified a gene encoding a novel soluble receptor belonging to the class II cytokine receptor family. This gene is positioned on chromosome 6 in the vicinity of the IFNGR1 gene in a head-to-tail orientation. The gene consists of six exons and encodes a 231-aa protein with a 21-aa leader sequence. The secreted mature protein demonstrates 34% amino acid identity to the extracellular domain of the IL-22R1 chain. Cross-linking experiments demonstrate that the protein binds IL-22 and prevents binding of IL-22 to the functional cell surface IL-22R complex, which consists of two subunits, the IL-22R1 and the IL-10R2c chains. Moreover, this soluble receptor, designated IL-22-binding protein (BP), is capable of neutralizing IL-22 activity. In the presence of the IL-22BP, IL-22 is unable to induce Stat activation in IL-22-responsive human lung carcinoma A549 cells. IL 22BP also blocked induction of the suppressors of cytokine signaling-3 (SOCS-3) gene expression by IL-22 in HepG2 cells. To further evaluate IL-22BP action, we used hamster cells expressing a modified IL-22R complex consisting of the intact IL-10R2c and the chimeric IL-22R1/gammaR1 receptor in which the IL-22R1 intracellular domain was replaced with the IFN-gammaR1 intracellular domain. In these cells, IL-22 activates biological activities specific for IFN-gamma, such as up-regulation of MHC class I Ag expression. The addition of IL-22BP neutralizes the ability of IL-22 to induce Stat activation and MHC class I Ag expression in these cells. Thus, the soluble receptor designated IL-22BP inhibits IL-22 activity by binding IL-22 and blocking its interaction with the cell surface IL-22R complex. PMID- 11390455 TI - Differential regulation of chemokine gene expression by 15-deoxy-delta 12,14 prostaglandin J2. AB - Ligands for peroxisome proliferator-activated receptor gamma (PPARgamma), such as 15-deoxy-Delta(12,14)PGJ2 (15d-PGJ2) have been proposed as a new class of antiinflammatory compounds with possible clinical applications. As there is some controversy over the inhibitory effects of 15d-PGJ2 on chemokine gene expression, we investigated whether 15d-PGJ2 itself affected chemokine gene expression in human monocytes/macrophages and two monocytic cell lines. Here we demonstrate that the 15d-PGJ2 can induce IL-8 gene expression. In contrast, monocyte chemoattractant protein-1 gene expression was suppressed by 15d-PGJ2, while the expression of RANTES was unaltered. Furthermore, concomitant treatment of monocytes/macrophages with 15d-PGJ2 (2.5 x 10(-6) M) potentiated LPS-induced gene expression of IL-8 mRNA, but suppressed PMA-induction of IL-8 mRNA. In addition, treatment of U937 and THP-1 cells with 15d-PGJ2 also resulted in induction of IL 8 gene expression. Further studies demonstrated that 15d-PGJ2 regulated IL-8 gene expression via a ligand-specific and PPARgamma-dependent pathway. Our observations revealed a previous unappreciated function and mechanism of 15d-PGJ2 mediated regulation of cytokine gene expression in monocytes/macrophages. PMID- 11390456 TI - Opposite regulation of tissue factor expression by calcineurin in monocytes and endothelial cells. AB - Tissue factor (TF), the primary initiator of blood coagulation with structural homology to the cytokine receptor family, has been implicated in various vascular processes including metastasis, angiogenesis, and atherosclerosis. Within the vasculature, monocytes and endothelial cells (EC) can be activated to synthesize TF depending on the induction of NF-kappaB. Despite the undisputed value of cyclosporin A (CsA) as an immunosuppressant, problems have emerged due to induction of vascular changes by a poorly understood mechanism. We demonstrate that CsA has opposite effects on TF gene expression, inhibiting NF-kappaB mediated TF gene transcription in monocytes but enhancing it in EC. To test whether CsA binding proteins (cyclophilins) can mediate these CsA effects we used a nonimmunosuppressant analog of CsA that binds to cyclophilins but does not inhibit the Ca2+/calmodulin-dependent phosphatase calcineurin (Cn). This drug lacked regulatory function for NF-kappaB and TF expression suggesting that Cn is responsible for the inverse gene regulation. The key function of Cn was supported by experiments demonstrating that other phosphatase inhibitors also either positively or negatively regulated NF-kappaB in monocytes and EC. Calcineurin was demonstrated to regulate NF-kappaB activation at the level of IkappaBalpha degradation, because agonist-induced phosphorylation and subsequent degradation of IkappaBalpha is prevented by Cn inhibitors in monocytes but enhanced in EC. These data identify Cn as an opposite regulator in generating transcriptionally active NF-kappaB, and they confirm the presumption that the ability of Cn to participate in NF-kappaB transactivation is not T cell specific. PMID- 11390457 TI - Augmentation of RANTES-induced extracellular signal-regulated kinase mediated signaling and T cell adhesion by elastase-treated fibronectin. AB - T cells migrating across extracellular matrix (ECM) barriers toward their target, the inflammatory site, should respond to chemoattractant cytokines and to the degradation of ECM by specific enzymes. In this study, we examined the effects of RANTES and ECM proteins treated with human leukocyte elastase on T cell activation and adhesion to the ECM. We found that human peripheral blood T cells briefly suspended with RANTES (0.1-100 ng/ml) had increased phosphorylation of their intracellular extracellular signal-regulated kinase (ERK), a mitogen activated protein kinase involved in the activation of several intracellular downstream effector molecules implicated in cell adhesion and migration. Consequently, a small portion (12-20%) of the responding cells adhered to fibronectin (FN). However, when the T cells were exposed to RANTES in the presence of native immobilized FN, laminin, or collagen type I, ERK phosphorylation was partially inhibited, suggesting that this form of the ECM proteins can down-regulate RANTES-induced intracellular signaling. In contrast, when the T cells were exposed to RANTES in the presence of elastase-treated immobilized FN, but not to elastase-treated laminin, ERK phosphorylation was markedly increased. Furthermore, a large percentage (30%) of RANTES-activated T cells adhered to the enzymatically treated FN in a beta1 integrin-dependent fashion. Thus, while migrating along chemotactic gradients within the ECM, T cells can adapt their adhesive performance according to the level of cleavage induced by enzymes to the matrix. PMID- 11390458 TI - An essential role of the NF-kappa B/Toll-like receptor pathway in induction of inflammatory and tissue-repair gene expression by necrotic cells. AB - Tissue damage induced by infection or injury can result in necrosis, a mode of cell death characterized by induction of an inflammatory response. In contrast, cells dying by apoptosis do not induce inflammation. However, the reasons for underlying differences between these two modes of cell death in inducing inflammation are not known. Here we show that necrotic cells, but not apoptotic cells, activate NF-kappaB and induce expression of genes involved in inflammatory and tissue-repair responses, including neutrophil-specific chemokine genes KC and macrophage-inflammatory protein-2, in viable fibroblasts and macrophages. Intriguingly, NF-kappaB activation by necrotic cells was dependent on Toll-like receptor 2, a signaling pathway that induces inflammation in response to microbial agents. These results have identified a novel mechanism by which cell necrosis, but not apoptosis, can induce expression of genes involved in inflammation and tissue-repair responses. Furthermore, these results also demonstrate that the NF-kappaB/Toll-like receptor 2 pathway can be activated both by exogenous microbial agents and endogenous inflammatory stimuli. PMID- 11390459 TI - Mammalian and viral IL-10 enhance C-C chemokine receptor 5 but down-regulate C-C chemokine receptor 7 expression by myeloid dendritic cells: impact on chemotactic responses and in vivo homing ability. AB - The immunosuppressive and anti-inflammatory cytokine IL-10 inhibits the phenotypic and functional maturation of dendritic cells (DC) and has been reported to confer tolerogenic properties on these important professional APC. Here, we exposed murine bone marrow-derived myeloid DC to either mouse (m) or viral (v) IL-10 early during their in vitro generation in response to GM-CSF and IL-4. Both mIL-10 and vIL-10 down-regulated the expression of CCR7 mRNA determined by RT-PCR, while mIL-10 up-regulated the expression of CCR5 transcripts. These changes in CCR7 and CCR5 expression were associated with inhibition and augmentation, respectively, of DC chemotaxis toward their respective agonists, macrophage inflammatory proteins 3beta and 1alpha, while in vivo homing of DC from peripheral s.c. sites to secondary lymphoid tissue of syngeneic or allogeneic recipients was significantly impaired. Anti-mIL-10R mAb reversed the effects of mIL-10 on CCR expression and restored DC homing ability. Retroviral transduction of mIL-10- and vIL-10-treated DC to overexpress transgenic CCR7 partially restored the cells' lymphoid tissue homing ability in allogeneic recipients. However, CCR7 gene transfer did not reinstate the capacity of IL-10-treated DC to prime host naive T cells for ex vivo proliferative responses or Th1 cytokine (IFN-gamma) production in response to rechallenge with (donor) alloantigen. These findings suggest that in addition to their capacity to subvert DC maturation/function and confer tolerogenic potential on these cells, mIL-10 and vIL-10 regulate DC migratory responses via modulation of CCR expression. PMID- 11390460 TI - The frequency of high avidity T cells determines the hierarchy of determinant spreading. AB - Autoimmunity often spreads in a predefined pattern during the progression of T cell-mediated autoimmune diseases. This progression has been well described in animal models and in man, but the basis for this phenomenon is little understood. To gain insight into the factors that determine this spreading hierarchy, we characterized the binding affinity of a panel of beta cell-autoantigenic peptides to I-Ag7, as well as the precursor frequency, functional avidity, and phenotype of the T cells that recognize these peptides in type 1 diabetes-prone nonobese diabetic mice. We observed that autoimmunity gradually spreads from a beta cell determinant, which had the largest precursor pool of high avidity T cells, to beta cell determinants with progressively smaller and lower avidity T cell precursor pools. This correlation between the sequential development of spontaneous T cell autoimmunity and the frequency and avidity of autoantigen reactive T cells suggests that the extent to which T cells were negatively selected by the self-determinants is the key factor determining the spreading hierarchy. PMID- 11390461 TI - Pharmacokinetic differences between a T cell-tolerizing and a T cell-activating peptide. AB - Vaccination with a peptide representing a CTL epitope from the human papillomavirus (HPV)16 E7 protein induces a specific CTL response that prevents the outgrowth of HPV16 E7-expressing tumors. In contrast, vaccination with a peptide encoding an adenovirus type 5 (Ad5) E1A CTL epitope results in CTL tolerance and enhanced growth of an Ad5 E1A-expressing tumor. It is unclear why these peptides induce such opposite effects. To determine whether a difference in pharmacokinetics can explain the functional contrasts, tritiated Ad5 E1A and HPV16 E7 peptides were injected into mice. Results show that the tolerizing peptide spread through the body 16 times faster than the activating peptide and was cleared at least 2 times faster. The HPV16 E7 peptide kinetics correlated with the kinetics of HPV16 E7-specific CTL induction. In contrast, Ad5 E1A peptide injection resulted in physical deletion of preexisting Ad5 E1A-specific CTLs within 24 h after injection. This tolerization occurred at the time when the peptide reached its maximum peptide concentration in the organs. These data suggest that ubiquitous expression of the tolerizing Ad5 E1A peptide within a short period of time causes activation-induced cell death of Ad5 E1A-specific CTLs. Therefore, information on the pharmacokinetics of peptides is vital for the safety and efficacy of peptide-based vaccines. PMID- 11390462 TI - Tolerance to cyclosporin A-induced autologous graft-versus-host disease is mediated by a CD4+CD25+ subset of recent thymic emigrants. AB - Our previous studies revealed that both the autoeffector and immunoregulatory T cells in cyclosporin A (CSA)-induced autologous graft-vs-host disease are recent thymic emigrants (RTEs). The autoeffector cells appear in and are released from the thymus during the first week of CSA treatment, whereas the immunoregulatory thymocytes appear during the second week but are not released until several days after cessation of CSA treatment. In the present study, the antigenic phenotypes of these functional T cell subsets were determined by immunomagnetic separation and flow immunocytometric analysis. During CSA wk 1, the autoeffector T cells in both the thymus and lymph node (LN) expressed a CD4+8+ double-positive (DP) phenotype, after which those in the LN became CD8 single positive (SP). Timed thymectomy experiments confirmed that the CD8-SP autoeffector T cells in LN originated from these DP RTEs. During CSA wk 2, the immunoregulatory thymocytes also displayed a DP phenotype. However, they were not exported to the periphery until several days after CSA treatment had been interrupted and they had acquired a CD4-SP phenotype. In LN, these immunoregulatory RTEs expressed the CD25+ marker characteristic of anergic/suppressor T cells. Cell separation and mixing experiments demonstrated that the autoeffector T cells persist in LN after cessation of CSA treatment, but their activity is not detectable in the presence of recently exported CD4+ T cells. Hence, the results indicate that tolerance to CSA-induced autologous graft-vs-host disease is actively mediated by CD25+CD4+ RTEs that suppress the function of CD8 autoeffector T cells. PMID- 11390463 TI - Sanglifehrin A, a novel cyclophilin-binding compound showing immunosuppressive activity with a new mechanism of action. AB - We report here on the characterization of the novel immunosuppressant Sanglifehrin A (SFA). SFA is a representative of a class of macrolides produced by actinomycetes that bind to cyclophilin A (CypA), the binding protein of the fungal cyclic peptide cyclosporin A (CsA). SFA interacts with high affinity with the CsA binding side of CypA and inhibits its peptidyl-prolyl isomerase activity. The mode of action of SFA is different from known immunosuppressive drugs. It has no effect on the phosphatase activity of calcineurin, the target of the immunosuppressants CsA and FK506 when complexed to their binding proteins CypA and FK binding protein, respectively. Moreover, its effects are independent of binding of cyclophilin. SFA inhibits alloantigen-stimulated T cell proliferation but acts at a later stage than CsA and FK506. In contrast to these drugs, SFA does not affect IL-2 transcription or secretion. However, it blocks IL-2 dependent proliferation and cytokine production of T cells, in this respect resembling rapamycin. SFA inhibits the proliferation of mitogen-activated B cells, but, unlike rapamycin, it has no effect on CD154/IL-4-induced Ab synthesis. The activity of SFA is also different from that of other known late acting immunosuppressants, e.g., mycophenolate mofetil or brequinar, as it does not affect de novo purine and pyrimidine biosynthesis. In summary, we have identified a novel immunosuppressant, which represents, in addition to CsA, FK506 and rapamycin, a fourth class of immunophilin-binding metabolites with a new, yet undefined mechanism of action. PMID- 11390464 TI - The P2Y11 receptor mediates the ATP-induced maturation of human monocyte-derived dendritic cells. AB - Recently, it has been shown that ATP and TNF-alpha synergize in the activation and maturation of human dendritic cells (DC); the effect of ATP was reproduced by hydrolysis-resistant derivatives of ATP and was blocked by suramin, suggesting the involvement of a P2 receptor, but the particular subtype involved was not identified. In this report we confirm that ATP and various derivatives synergize with TNF-alpha and LPS to induce the maturation of human monocyte-derived DC, as revealed by up-regulation of the CD83 marker and the secretion of IL-12. The rank order of potency of various analogs (AR-C67085 > adenosine 5'-O-(3 thiotriphosphate) = 2'- and 3'-O-(4-benzoyl-benzoyl) ATP > ATP > 2-methylthio ATP) was close to that of the recombinant human P2Y11 receptor. Furthermore, these compounds activated cAMP production in DC, in a xanthine-insensitive way, consistent with the involvement of the P2Y11 receptor, which among P2Y subtypes has the unique feature of being dually coupled to phospholipase C and adenylyl cyclase activation. The involvement of the P2Y11/cAMP/protein kinase A signaling pathway in the nucleotide-induced maturation of DC is supported by the inhibitory effect of H89, a protein kinase A inhibitor. Taken together, our results demonstrate that ATP activates DC through stimulation of the P2Y11 receptor and subsequent increase in intracellular cAMP. PMID- 11390465 TI - In vivo-matured Langerhans cells continue to take up and process native proteins unlike in vitro-matured counterparts. AB - We have been able to identify the cell subset derived from Langerhans cells in the total dendritic cell population of the peripheral lymph node and hence to follow their trafficking under normal physiological conditions as well as upon skin irritation. As expected, the rapid mobilization of Langerhans cells triggered by inflammatory signals into the draining lymph node correlated with an up-regulation of costimulatory molecules and with an enhanced immunostimulatory capacity. Surprisingly, however, these cells, instead of shutting down, maintain the capacity to capture and process protein Ags during the couple of days they stay alive in stark contrast to in vitro-matured dendritic cells. PMID- 11390466 TI - Progesterone up-regulates anandamide hydrolase in human lymphocytes: role of cytokines and implications for fertility. AB - Physiological concentrations of progesterone stimulate the activity of the endocannabinoid-degrading enzyme anandamide hydrolase (fatty acid amide hydrolase, FAAH) in human lymphocytes. At the same concentrations, the membrane impermeant conjugate of progesterone with BSA was ineffective, suggesting that binding to an intracellular receptor was needed for progesterone activity. Stimulation of FAAH occurred through up-regulation of gene expression at transcriptional and translational level, and was partly mediated by the Th2 cytokines. In fact, lymphocyte treatment with IL-4 or with IL-10 had a stimulating effect on FAAH, whereas the Th1 cytokines IL-12 and IFN-gamma reduced the activity and the protein expression of FAAH. Human chorionic gonadotropin or cortisol had no effect on FAAH activity. At variance with FAAH, the lymphocyte anandamide transporter and cannabinoid receptors were not affected by treatment with progesterone or cytokines. Good FAAH substrates such as anandamide and 2 arachidonoylglycerol inhibited the release of leukemia-inhibitory factor from human lymphocytes, but N-palmitoylethanolamine, a poor substrate, did not. A clinical study performed on 100 healthy women showed that a low FAAH activity in lymphocytes correlates with spontaneous abortion, whereas anandamide transporter and cannabinoid receptors in these cells remain unchanged. These results add the endocannabinoids to the hormone-cytokine array involved in the control of human pregnancy. PMID- 11390467 TI - Production of TNF-alpha by human V gamma 9V delta 2 T cells via engagement of Fc gamma RIIIA, the low affinity type 3 receptor for the Fc portion of IgG, expressed upon TCR activation by nonpeptidic antigen. AB - Human lymphocytes expressing the gammadelta TCR represent a minor T cell subpopulation found in blood. The majority of these cells express Vgamma9Vdelta2 determinants and respond to nonpeptidic phosphoantigens. Several studies have shown that, in vivo, the percentage of Vgamma9Vdelta2 T cells dramatically increases during pathological infection, leading to the hypothesis that they play an important role in the defense against pathogens. However, the specific mechanisms involved in this response remain poorly understood. It has been established that Vgamma9Vdelta2 T cells display potent cytotoxic activity against virus-infected and tumor cells, thereby resembling NK cells. In this study, we show that, upon stimulation by nonpeptidic Ags, Vgamma9Vdelta2 T cells express FcgammaRIIIA (CD16), a receptor that is constitutively expressed on NK cells. CD16 appears to be an activation Ag for Vgamma9Vdelta2 T cells. Indeed, ligation of CD16 on Vgamma9Vdelta2 T cells leads to TNF-alpha production. This TNF-alpha production, which is dependent (like that induced via the TCR-CD3 complex) on the activation of the p38 and extracellular signal-regulated kinase-2 mitogen activated protein kinases, can be modulated by CD94 NK receptors. Therefore, it appears that Vgamma9Vdelta2 T cells can be physiologically activated by two sequential steps via two different cell surface Ags: the TCR-CD3 complex and the FcgammaRIIIA receptor, which are specific cell surface Ags for T lymphocytes and NK cells, respectively. This strongly suggests that, in the general scheme of the immune response, Vgamma9Vdelta2 T cells represent an important subpopulation of cells that play a key role in the defense against invading pathogens. PMID- 11390468 TI - Differential survival of transferred CD8 T cells and host reconstitution depending on TCR avidity for host-expressed alloantigen. AB - We transferred naive alloreactive CD8 T cells from TCR transgenic mice to irradiated recipients expressing a partial (H-2Kbm8) or a full (H-2Kb) agonist alloantigen (alloAg). The consequences were strikingly distinct, resulting in acceleration of host lymphopoiesis in the former group, but in strong graft-vs host reaction, preventing host lymphocyte reconstitution in the latter group. This was correlated, respectively, with long-term persistence and with rapid disappearance of the transferred CD8 T cells. Analysis of transferred T cells showed that initial T cell expansion and modulation of expression of activation markers CD44 and CD62L, as well as induction of cytotoxic function, were similar in both groups. However, IL-2 production and subsequent up-regulation of CD25, early perforin-independent cytolysis, and early down-regulation of Bcl-2 expression were detected only in T cells transferred in hosts expressing full agonist alloAg. Expansion of transferred CD8 T cells was not dependent on either IL-2 or CD25 expression. This expansion could lead to either accelerated host reconstitution or to strong graft-vs-host, depending on the nature of the alloAg. Thus, the extent of Ag stimulation may be a crucial parameter in protocols of alloreactive T cell immunotherapy. PMID- 11390469 TI - CD45 function is regulated by an acidic 19-amino acid insert in domain II that serves as a binding and phosphoacceptor site for casein kinase 2. AB - In this study experiments were conducted to elucidate the physical/functional relationship between CD45 and casein kinase 2 (CK2). Immunoprecipitation experiments demonstrated that CK2 associates with CD45 and that this interaction is inducible upon Ag receptor cross-linking in B and T cell lines as well as murine thymocytes and splenic B cells. However, yeast two-hybrid analysis failed to demonstrate a physical interaction between the individual CK2 alpha, alpha', or beta subunits and CD45. In contrast, a yeast three-hybrid assay in which either CK2 alpha and beta or alpha' and beta subunits were coexpressed with the cytoplasmic domain of CD45, demonstrated that both CK2 subunits are necessary for the interaction with CD45. Experiments using the yeast three-hybrid assay also revealed that a 19-aa acidic insert in domain II of CD45 mediates the physical interaction between CK2 and CD45. Structure/function experiments in which wild type or mutant CD45RA and CD45RO isoforms were expressed in CD45-deficient Jurkat cells revealed that the 19-aa insert is important for optimal CD45 function. The ability of both CD45RA and CD45RO to reconstitute CD3-mediated signaling based on measurement of calcium mobilization and mitogen-activated protein kinase activation was significantly decreased by deletion of the 19-aa insert. Mutation of four serine residues within the 19-aa insert to alanine affected CD45 function to a similar extent compared with that of the deletion mutants. These findings support the hypothesis that a physical interaction between the CD45 cytoplasmic domain and CK2 is important for post-translational modification of CD45, which, in turn, regulates its catalytic function. PMID- 11390471 TI - Quantitative relationship between MHC class II-superantigen complexes and the balance of T cell activation versus death. AB - The binding of bacterial superantigens (SAgs) is profoundly affected by the nature of the MHC class II-associated antigenic peptide. It was proposed that this limitation in the density of SAgs displayed at the surface of APCs is important for efficient TCR serial triggering as well as for preventing apoptosis of the responding T lymphocytes. Here, we have addressed quantitatively the size of this SAg-receptive pool of HLA-DR molecules that are available to bind and present staphylococcal enterotoxin A (SEA) at the surface of B lymphocytes. Our binding curves, depletion experiments, and quantitative immunoprecipitations show that about half the HLA-DR class II molecules on B cells are refractory to SEA binding. Yet, as compared with typical nominal Ags, an unusually high amount of class II-SAg complexes can be presented to T cells. This characteristic appears to be necessary for SAg-induced T cell apoptosis. When <0.3% of the total cell surface MHC class II molecules are occupied by SEA, T cells undergo a normal sequence of early activation events. However, presentation of a ligand density beyond this threshold results in T cell activation that is readily aborted by apoptosis but only after a few cell divisions. Thus, we confirm the existence of MHC class II subsets that are structurally unable to present SEA and provide a quantitative framework to account for the ability of bacterial SAgs to induce peripheral activation vs tolerance in the host. PMID- 11390470 TI - Regulation of NK cell-mediated cytotoxicity by the adaptor protein 3BP2. AB - Stimulation of lymphocytes through multichain immune recognition receptors activates multiple signaling pathways. Adaptor proteins play an important role in integrating these pathways by their ability to simultaneously bind multiple signaling components. Recently, the 3BP2 adaptor protein has been shown to positively regulate the transcriptional activity of T cells. However, the mechanisms by which signaling components are involved in this regulation remain unclear, as does a potential role for 3BP2 in the regulation of other cellular functions. Here we describe a positive regulatory role for 3BP2 in NK cell mediated cytotoxicity. We also identify p95(vav) and phospholipase C-gamma isoforms as binding partners of 3BP2. Our results show that tyrosine-183 of 3BP2 is specifically involved in this interaction and that this residue critically influences 3BP2-dependent function. Therefore, 3BP2 regulates NK cell-mediated cytotoxicity by mobilizing key downstream signaling effectors. PMID- 11390472 TI - Processing of immunosuppressive pro-TGF-beta 1,2 by human glioblastoma cells involves cytoplasmic and secreted furin-like proteases. AB - TGF-beta is a putative mediator of immunosuppression associated with malignant glioma and other types of cancer. Subtilisin-like proprotein convertases such as furin are thought to mediate TGF-beta processing. Here we report that human malignant glioma cell lines express furin mRNA and protein, exhibit furin-like protease (FLP) activity, and release active furin into the cell culture supernatant. FLP activity is not modulated by exogenous TGF-beta or neutralizing TGF-beta Abs. Exposure of LN-18 and T98G glioma cell lines to the furin inhibitor, decanoyl-Arg-Val-Lys-Arg-chloromethylketone, inhibits processing of the TGF-beta1 and TGF-beta2 precursor molecules and, consequently, the release of mature bioactive TGF-beta molecules. Ectopic expression of PDX, a synthetic antitrypsin analog with antifurin activity, in the glioma cells inhibits FLP activity, TGF-beta processing, and TGF-beta release. Thus, subtilisin-like proprotein convertases may represent a novel target for the immunotherapy of malignant glioma and other cancers or pathological conditions characterized by enhanced TGF-beta bioactivity. PMID- 11390473 TI - Extrathymic development of V alpha 11 T cells in placenta during pregnancy and their possible physiological role. AB - The molecular and cellular mechanisms of the feto-maternal immune responses in the placenta in connection with natural abortion remain unclear. In this report we provide evidence that V(alpha11) T cells developed in the placenta may be responsible for the induction of natural abortion. The majority of V(alpha11) TCRs detected during pregnancy showed a consensus motif in the CDR3 region, similar to that of anti-GM3 TCR clones, and were of maternal origin. V(alpha11) TCRs were found in the middle to late stages of gestation due to de novo generation in the placenta, not to migration from the maternal side, as evidenced by the significant increases in the out-of-frame V(alpha11) TCR mRNA and the copy number of circular DNA generated by V(alpha11) gene rearrangements. Furthermore, administration of anti-V(alpha11) Ab to pregnant mice resulted in a significant decrease in the incidence of fetal demise, suggesting that V(alpha11) T cells detected in the placenta develop extrathymically and are involved in natural abortion. PMID- 11390474 TI - Novel G protein-coupled responses in leukocytes elicited by a chemotactic bacteriophage displaying a cell type-selective binding peptide. AB - Recently, we identified a neutrophil-binding phage displaying a novel peptide motif, GPNLTGRW. It was determined that this peptide, when displayed on bacteriophage (FGP phage), elicits a transient increase in cytosolic calcium. Here, we show that FGP phage stimulate neutrophil chemotaxis and induce a pertussis toxin-sensitive rise in cytosolic calcium in monocytes as well as in neutrophils. In contrast to the calcium response elicited by classical chemoattractants fMLP and IL-8, the FGP phage-elicited response in neutrophils is dependent on extracellular calcium and is mediated by receptor-activated, divalent cation channels. Consistent with G protein-coupled receptor signaling, FGP phage effect homologous and reciprocal heterologous desensitization with fMLP and IL-8-stimulated calcium responses. Like non-G protein-coupled responses, the FGP-elicited calcium transient is abolished with phosphoinositide-3-kinase inactivation. Nonetheless, specific binding of GTP to neutrophil membranes follows stimulation with FGP phage, further supporting involvement of G proteins. However, FGP phage neither bind to nor elicit a calcium response from transfectant cells harboring known candidate G protein-coupled receptors. These data together suggest that the elicited responses are mediated by a novel G protein-coupled receptor or represent novel responses of a known receptor. PMID- 11390475 TI - Recognition of HLA-Cw4 but not HLA-Cw6 by the NK cell receptor killer cell Ig like receptor two-domain short tail number 4. AB - NK cells are cytotoxic to virus-infected and tumor cells that have lost surface expression of class I MHC proteins. Target cell expression of class I MHC proteins inhibits NK cytotoxicity through binding to inhibitory NK receptors. In contrast, a similar family of activating NK receptors, characterized by the presence of a charged residue in their transmembrane portion and a truncated cytoplasmic tail, augment lysis by NK cells when ligated by an appropriate class I MHC protein. However, the class I MHC specificity of many of these activating NK receptors is still unknown. Here, we show enhanced lysis of HLA-Cw4 but not HLA-Cw6-expressing cells, by a subset of NK clones. This subset may express killer cell Ig-like receptor two-domain short tail number 4 (KIR2DS4), as suggested by staining with various mAb. It is still possible, however, that these clones may express receptors other than KIR2DS4 that might recognize HLA-Cw4. Binding of KIR2DS4-Ig fusion protein to cells expressing HLA-Cw4 but not to those expressing HLA-Cw6 was also observed. The binding of KIR2DS4-Ig to HLA-Cw4 is weaker than that of killer cell Ig-like receptor two-domain long tail number 1 (KIR2DL1)-Ig fusion protein; however, such weak recognition is capable of inhibiting lysis by an NK transfectant expressing a chimeric molecule of KIR2DS4 fused to the transmembrane and cytoplasmic portion of KIR2DL1. Residue alpha14 is shown to be important in the KIR2DS4 binding to HLA-Cw4. Implications of the role of the activating NK receptors in immunosurveillance are discussed. PMID- 11390476 TI - Factors affecting the efficiency of CD8+ T cell cross-priming with exogenous antigens. AB - Processing of exogenous protein Ags by APC leads predominantly to presentation of peptides on class II MHC and, thus, stimulation of CD4+ T cell responses. However, "cross-priming" can also occur, whereby peptides derived from exogenous Ags become displayed on class I MHC molecules and stimulate CD8+ T cell responses. We compared the efficiency of cross-priming with exogenous proteins to use of peptide Ags in human whole blood using a flow cytometry assay to detect T cell intracellular cytokine production. CD8+ T cell responses to whole CMV proteins were poorly detected (compared with peptide responses) in most CMV seropositive donors. Such responses could be increased by using higher doses of Ag than were required to achieve maximal CD4+ T cell responses. A minority of donors displayed significantly more efficient CD8+ T cell responses to whole protein, even at low Ag doses. These responses were MHC class I-restricted and dependent upon proteosomal processing, indicating that they were indeed due to cross-priming. The ability to efficiently cross-prime was not a function of the number of dendritic cells in the donor's blood. Neither supplementation of freshly isolated dendritic cells nor use of cultured, Ag-pulsed dendritic cells could significantly boost CD8 responses to whole-protein Ags in poorly cross priming donors. Interestingly, freshly isolated monocytes performed almost as well as dendritic cells in inducing CD8 responses via cross-priming. In conclusion, the efficiency of cross-priming appears to be poor in most donors and is dependent upon properties of the individual's APC and/or T cell repertoire. It remains unknown whether cross-priming ability translates into any clinical advantage in ability to induce CD8+ T cell responses to foreign Ags. PMID- 11390477 TI - Stat6 is necessary and sufficient for IL-4's role in Th2 differentiation and cell expansion. AB - IL-4 plays a critical role in the differentiation of TCR-stimulated naive CD4 T cells to the Th2 phenotype. In response to IL-4, the IL-4R activates a set of phosphotyrosine binding domain-containing proteins, including insulin receptor substrate 1/2, Shc, and IL-4R interacting protein, as well as Stat6. Stat6 has been shown to be required for Th2 differentiation. To determine the roles of the phosphotyrosine binding adaptors in Th2 differentiation, we prepared a retrovirus containing a mutant of the human (h)IL-4R alpha-chain, Y497F, which is unable to recruit these adaptors. The mutant hIL-4Ralpha, as well as the wild-type (WT) hIL 4Ralpha, was introduced into naive CD4 T cells. Upon hIL-4 stimulation, Y497F worked as well as the WT hIL-4Ralpha in driving Th2 differentiation, as measured by Gata3 up-regulation and IL-4 production. Furthermore, IL-4-driven cell expansion was also normal in the cells infected with Y497F, although cells infected with Y497F were not capable of phosphorylating insulin receptor substrate 2. These results suggest that the signal pathway mediated by Y497 is dispensable for both IL-4-driven Th2 differentiation and cell expansion. Both WT and Y497F hIL-4Ralpha lose the ability to drive Th2 differentiation and cell expansion in Stat6-knockout CD4 T cells. A constitutively activated form of Stat6 introduced into CD4 T cells resulted in both Th2 differentiation and enhanced cell expansion. Thus, activated Stat6 is necessary and sufficient to mediate both IL-4-driven Th2 differentiation and cell expansion in CD4 T cells. PMID- 11390478 TI - A role for TGF-beta in the generation and expansion of CD4+CD25+ regulatory T cells from human peripheral blood. AB - An elusive goal in transplanting organs across histocompatibility barriers has been the induction of specific tolerance to avoid graft rejection. A considerable body of evidence exists that the thymus produces regulatory T cells that suppress the response of other T cells to antigenic stimulation. We report that TGF-beta can induce certain CD4+ T cells in the naive (CD45RA+RO-) fraction in human peripheral blood to develop powerful, contact-dependent suppressive activity that is not antagonized by anti-TGF-beta or anti-IL-10 mAbs. The costimulatory effects of TGF-beta on naive CD4+ T cells up-regulated CD25 and CTLA-4 expression, increased their transition to the activated phenotype, but decreased activation induced apoptosis. Suppressive activity was concentrated in the CD25+ fraction. These CD4+CD25+ regulatory cells prevented CD8+ T cells from proliferating in response to alloantigens and from becoming cytotoxic effector cells. Moreover, these regulatory cells exerted their suppressive activities in remarkably low numbers and maintained these effects even after they are expanded. Once activated, their suppressive properties were Ag nonspecific. Although <1% of naive CD4+ T cells expressed CD25, depletion of this subset before priming with TGF-beta markedly decreased the generation of suppressive activity. This finding suggests that CD4+CD25+ regulatory T cells induced ex vivo are the progeny of thymus-derived regulatory T cells bearing a similar phenotype. The adoptive transfer of these regulatory T cells generated and expanded ex vivo has the potential to prevent rejection of allogeneic organ grafts. PMID- 11390479 TI - Evidence for epigenetic mechanisms that silence both basal and immune-stimulated transcription of the IL-8 gene. AB - It is becoming increasingly clear that epigenetic silencing of gene transcription plays a critical role in the regulation of gene expression in many biological processes. Tight regulation of immunomodulatory substances that are important for the initiation of the inflammatory cascade, such as chemoattractive cytokines, is essential to prevent initiation of unrestrained immune activation. Using the Caco 2 intestinal cell line as a model, we reveal two distinctly different mechanisms by which the gene for the neutrophil chemoattractive cytokine IL-8 is silenced. Nuclear run-on studies, as well as stably transfected reporter and marked minigene constructs, demonstrate that cellular differentiation inhibits immune activated transcription of the IL-8 gene, a mechanism that is dependent on histone deacetylase activity. Unexpectedly, this silencing mechanism does not involve previously described regulatory elements in the IL-8 promoter but rather cis-acting regions located at a distance from the IL-8 gene locus. Genomic elements distant to the immediate IL-8 locus are also required to silence aberrant basal transcriptional activity of the IL-8 promoter in the absence of immune activation. However, in this case, silencing occurs in a histone deacetylase-independent fashion. These findings were confirmed in transgenic mice in which, in the absence of these elements, aberrant IL-8 gene activity was present primarily in the intestinal tract. Epigenetic silencing of cytokine gene transcription through distant genomic elements is an important level of gene regulation that may be relevant to the pathogenesis of immunologic disease states. PMID- 11390480 TI - Differential expression of inducible costimulator-ligand splice variants: lymphoid regulation of mouse GL50-B and human GL50 molecules. AB - The process of immunological costimulation between APC and T cells is mediated by protein ligand:receptor interactions. To date, costimulatory receptors known to be expressed by T cells include the structurally related proteins CD28 and the inducible costimulator (ICOS). The ligands to human and mouse ICOS, human GL50 (hGL50), and mouse GL50 (mGL50) were recently cloned and demonstrated to have sequence similarity to the CD28 ligands B7-1 and B7-2. Examination of mGL50 cDNA transcripts by 3'RACE revealed an alternatively spliced form, mGL50-B, that encoded a protein product with a divergent 27-aa intracellular domain. Both mGL50 and mGL50-B-transfected cells exhibited binding to human and mouse ICOS-Ig fusion protein, indicating that the alternate cytoplasmic domain of mGL50-B does not interfere with extracellular interactions with ICOS receptor. Flow cytometric and RT-PCR analysis of BALB/c and RAG1(-/-) mice splenocytes demonstrate that freshly isolated B cells, T cells, macrophages, and dendritic cells express both splice variant forms of ICOS ligand. Comparative analyses with the human ICOS ligand splice variants hGL50 and B7-H2 indicate that differential splicing at the junction of cytoplasmic exon 6 and exon 7 may be a common method by which GL50 ICOS immunological costimulatory processes are regulated in vivo. PMID- 11390481 TI - Proteomic analysis of dendritic cell-derived exosomes: a secreted subcellular compartment distinct from apoptotic vesicles. AB - Dendritic cells constitutively secrete a population of small (50-90 nm diameter) Ag-presenting vesicles called exosomes. When sensitized with tumor antigenic peptides, dendritic cells produce exosomes, which stimulate anti-tumor immune responses and the rejection of established tumors in mice. Using a systematic proteomic approach, we establish the first extensive protein map of a particular exosome population; 21 new exosomal proteins were thus identified. Most proteins present in exosomes are related to endocytic compartments. New exosomal residents include cytosolic proteins most likely involved in exosome biogenesis and function, mainly cytoskeleton-related (cofilin, profilin I, and elongation factor 1alpha) and intracellular membrane transport and signaling factors (such as several annexins, rab 7 and 11, rap1B, and syntenin). Importantly, we also identified a novel category of exosomal proteins related to apoptosis: thioredoxin peroxidase II, Alix, 14-3-3, and galectin-3. These findings led us to analyze possible structural relationships between exosomes and microvesicles released by apoptotic cells. We show that although they both represent secreted populations of membrane vesicles relevant to immune responses, exosomes and apoptotic vesicles are biochemically and morphologically distinct. Therefore, in addition to cytokines, dendritic cells produce a specific population of membrane vesicles, exosomes, with unique molecular composition and strong immunostimulating properties. PMID- 11390482 TI - Characterization of a pattern recognition protein, a masquerade-like protein, in the freshwater crayfish Pacifastacus leniusculus. AB - A multifunctional masquerade-like protein has been isolated, purified, and characterized from hemocytes of the freshwater crayfish, Pacifastacus leniusculus. It was isolated by its Escherichia coli binding property, and it binds to formaldehyde-treated Gram-negative bacteria as well as to yeast, Saccharomyces cerevisiae, whereas it does not bind to formaldehyde-fixed Gram positive bacteria. The intact masquerade (mas)-like protein is present in crayfish hemocytes as a heterodimer composed of two subunits with molecular masses of 134 and 129 kDa. Under reducing conditions the molecular masses of the intact proteins are not changed. After binding to bacteria or yeast cell walls, the mas-like protein is processed by a proteolytic enzyme. The 134 kDa of the processed protein yields four subunits of 65, 47, 33, and 29 kDa, and the 129-kDa protein results in four subunits of 63, 47, 33, and 29 kDa in 10% SDS-PAGE under reducing conditions. The 33-kDa protein could be purified by immunoaffinity chromatography using an Ab to the C-terminal part of the mas-like protein. This subunit of the mas-like protein has cell adhesion activity, whereas the two intact proteins, 134 and 129 kDa, have binding activity to LPSs, glucans, Gram negative bacteria, and yeast. E. coli coated with the mas-like protein were more rapidly cleared in crayfish than only E. coli, suggesting this protein is an opsonin. Therefore, the cell adhesion and opsonic activities of the mas-like protein suggest that it plays a role as an innate immune protein. PMID- 11390484 TI - Augmentation in expression of activation-induced genes differentiates memory from naive CD4+ T cells and is a molecular mechanism for enhanced cellular response of memory CD4+ T cells. AB - In an attempt to understand the molecular basis for the immunological memory response, we have used cDNA microarrays to measure gene expression of human memory and naive CD4+ T cells at rest and after activation. Our analysis of 54,768 cDNA clones provides the first glimpse into gene expression patterns of memory and naive CD4+ T cells at the genome-scale and reveals several novel findings. First, memory and naive CD4+ T cells expressed similar numbers of genes at rest and after activation. Second, we have identified 14 cDNA clones that expressed higher levels of transcripts in memory cells than in naive cells. Third, we have identified 135 (130 known genes and 5 expressed sequence tags) up regulated and 68 (42 known genes and 26 expressed sequence tags) down-regulated cDNA clones in memory CD4+ T after in vitro stimulation with anti-CD3 plus anti CD28. Interestingly, the increase in mRNA levels of up-regulated genes was greater in memory than in naive CD4+ T cells after in vitro stimulation and was higher with anti-CD3 plus anti-CD28 than with anti-CD3 alone in both memory and naive CD4+ T cells. Finally, the changes in expression of actin and cytokine genes identified by cDNA microarrays were confirmed by Northern and protein analyses. Together, we have identified approximately 200 cDNA clones whose expression levels changed after activation and suggest that the level of expression of up-regulated genes is a molecular mechanism that differentiates the response of memory from naive CD4+ T cells. PMID- 11390483 TI - MHC class I recognition by NK receptors in the Ly49 family is strongly influenced by the beta 2-microglobulin subunit. AB - NK cell recognition of targets is strongly affected by MHC class I specific receptors. The recently published structure of the inhibitory receptor Ly49A in complex with H-2Dd revealed two distinct sites of interaction in the crystal. One of these involves the alpha1, alpha2, alpha3, and beta2-microglobulin (beta2m) domains of the MHC class I complex. The data from the structure, together with discrepancies in earlier studies using MHC class I tetramers, prompted us to study the role of the beta2m subunit in MHC class I-Ly49 interactions. Here we provide, to our knowledge, the first direct evidence that residues in the beta2m subunit affect binding of MHC class I molecules to Ly49 receptors. A change from murine beta2m to human beta2m in three different MHC class I molecules, H-2Db, H 2Kb, and H-2Dd, resulted in a loss of binding to the receptors Ly49A and Ly49C. Analysis of the amino acids involved in the binding of Ly49A to H-2Dd in the published crystal structure, and differing between the mouse and the human beta2m, suggests the cluster formed by residues Lys3, Thr4, Thr28, and Gln29, as a potentially important domain for the Ly49A-H-2Dd interaction. Another possibility is that the change of beta2m indirectly affects the conformation of distal parts of the MHC class I molecule, including the alpha1 and alpha2 domains of the heavy chain. PMID- 11390485 TI - Bcl-2 targets protein phosphatase 1 alpha to Bad. AB - The diverse forms of protein phosphatase 1 (PP1) in vivo result from the association of the catalytic subunit with different regulatory subunits. We recently have described that PP1alpha is a Ras-activated Bad phosphatase that regulates IL-2 deprivation-induced apoptosis. With the yeast two-hybrid system, GST fusion proteins, indirect immunofluorescence, and coimmunoprecipitation, we found that Bcl-2 interacts with PP1alpha and Bad. In contrast, Bad did not interact with 14-3-3 protein. Bcl-2 depletion decreased phosphatase activity and association of PP1alpha to Bad. Bcl-2 contains the RIVAF motif, analogous to the well characterized R/KXV/IXF consensus motif shared by most PP1-interacting proteins. This sequence is involved in the binding of Bcl-2 to PP1alpha. Disruption of Bcl-2/PP1alpha association strongly decreased Bcl-2 and Bad associated phosphatase activity and formation of the trimolecular complex. These results suggest that Bcl-2 targets PP1alpha to Bad. PMID- 11390486 TI - Role of chemokines and formyl peptides in pneumococcal pneumonia-induced monocyte/macrophage recruitment. AB - Host-derived chemoattractant factors are suggested to play crucial roles in leukocyte recruitment elicited by inflammatory stimuli in vitro and in vivo. However, in the case of acute bacterial infections, pathogen-derived chemoattractant factors are also present, and it has not yet been clarified how cross-talk between chemoattractant receptors orchestrates diapedesis of leukocytes in this context of complex chemoattractant arrays. To investigate the role of chemokine (host-derived) and formyl peptide (pathogen-derived) chemoattractants in leukocyte extravasation in life-threatening infectious diseases, we used a mouse model of pneumococcal pneumonia. We found an increase in mRNA expression of eight chemokines (RANTES, macrophage-inflammatory protein (MIP)-1alpha, MIP-1beta, MIP-2, IP-10, monocyte chemoattractant protein (MCP)-1, T cell activation 3, and KC) within the lungs during the course of infection. KC and MIP-2 protein expression closely preceded pulmonary neutrophil recruitment, whereas MCP-1 protein production coincided more closely than MIP-1alpha with the kinetics of macrophage infiltration. In situ hybridization of MCP-1 mRNA suggested that MCP-1 expression started at peribronchovascular regions and expanded to alveoli-facing epithelial cells and infiltrated macrophages. Interestingly, administration of a neutralizing Ab against MCP-1, RANTES, or MIP 1alpha alone did not prevent macrophage infiltration into infected alveoli, whereas combination of the three Abs significantly reduced macrophage infiltration without affecting neutrophil recruitment. The use of an antagonist to N-formyl peptides, N-t-Boc-Phe-D-Leu-Phe-D-Leu-Phe, reduced both macrophages and neutrophils significantly. These data demonstrate that a complex chemokine network is activated in response to pulmonary pneumococcal infection, and also suggest an important role for fMLP receptor in monocyte/macrophage recruitment in that model. PMID- 11390487 TI - The differential roles of LFA-1 and Mac-1 in host defense against systemic infection with Streptococcus pneumoniae. AB - Mice deficient in CD18, which lack all four CD11 integrins, have leukocytosis and increased susceptibility to bacterial infection. To determine the effect of deficiencies in LFA-1 (CD11a/CD18) or Mac-1 (CD11b/CD18) on host defense against systemic bacterial infection, knockout mice were inoculated i.p. with Streptococcus pneumoniae. Increased mortality occurred in both LFA-1(-/-) (15 of 17 vs 13 of 35 in wild type (WT), p < 0.01) and Mac-1(-/-) (17 of 34 vs 6 of 25, p < 0.01) mice. All deaths in LFA-1(-/-) mice occurred after 72 h, whereas most deaths in Mac-1(-/-) mice occurred within 24-48 h. At 24 h, 21 of 27 Mac-1(-/-) mice were bacteremic, vs 15 of 25 WT (p = 0.05); no difference was observed between LFA-1(-/-) and WT. Increased bacteria were recovered from Mac-1(-/-) spleens at 2 h (p = 0.03) and 6 h (p = 0.002) and from livers (p = 0.001) by 6 h. No difference was observed at 2 h in LFA-1(-/-) mice, but by 6 h increased bacteria were recovered from spleens (p = 0.008) and livers (p = 0.04). Baseline and peak leukocyte counts were similar between Mac-1(-/-) and WT, but elevated in LFA-1(-/-). At 8 h, peritoneal neutrophils were increased in Mac-1(-/-), but not significantly different in LFA-1(-/-). Histopathologically, at 24 h Mac-1(-/-) animals had bacteremia and lymphoid depletion, consistent with sepsis. LFA-1(-/-) mice had increased incidence of otitis media and meningitis/encephalitis vs WT at 72 and 96 h. Both Mac-1 and LFA-1 play important but distinct roles in host defense to S. pneumoniae. PMID- 11390488 TI - Immunotherapy of melanoma: a dichotomy in the requirement for IFN-gamma in vaccine-induced antitumor immunity versus adoptive immunotherapy. AB - The mechanism by which tumors are rejected following the adoptive transfer of tumor-specific T cells is not well characterized. Recent work has challenged the requirement for cytotoxicity mediated by either the perforin/granzyme or Fas/Fas ligand pathway in T cell-mediated tumor regression. Many reports, including ours, suggest that tumor-specific production of IFN-gamma is critical for T cell mediated tumor regression. However, in most of these studies the evidence to support the role for IFN-gamma is only indirect. We have directly examined the requirement for IFN-gamma using IFN-gamma knockout (GKO) mice. The results show an interesting dichotomy in the requirement for IFN-gamma: Antitumor immunity induced by active-specific immunotherapy (vaccination) required IFN-gamma, whereas adoptive immunotherapy did not. In GKO mice vaccination with the GM-CSF gene-modified B16BL6-D5 tumor (D5-G6) failed to induce protective immunity against parental D5 tumor. However, adoptive transfer of effector T cells from GKO mice cured 100% of GKO mice with established pulmonary metastases and induced long term antitumor immunity and depigmentation of skin. Furthermore, in vivo neutralization of IFN-gamma by mAb treatment or adoptive transfer into IFN-gamma receptor knockout mice failed to block the therapeutic efficacy of effector T cells generated from wild-type or perforin knockout mice. Analysis of regressing metastases revealed similar infiltrates of macrophages and granulocytes in both wild-type and GKO mice. These results indicate that in this adoptive immunotherapy model, neither a direct effect on the tumor nor an indirect effect of IFN-gamma through activation of myeloid or lymphoid cells is critical for therapeutic efficacy. PMID- 11390489 TI - Fc receptor-mediated phagocytosis makes a significant contribution to clearance of influenza virus infections. AB - Fc receptors for IgG expressed on macrophages and NK cells are important mediators of opsonophagocytosis and Ab-dependent cell-mediated cytotoxicity. Phagocyte-mediated opsonophagocytosis is pivotal for protection against bacteria, but its importance in recovery from infection with intracellular pathogens is unclear. We have now investigated the role of opsonophagocytosis in protection against lethal influenza virus infection by using FcR gamma(-/-) mice. Absence of the FcR gamma-chain did not affect the expression of IFN-gamma and IL-10 in the lungs and spleens after intranasal immunization with an influenza subunit vaccine. Titers of serum and respiratory Abs of the IgM, IgG1, IgG2a, and IgA isotypes in FcR gamma(-/-) mice were similar to levels seen in FcR gamma(+/+) mice. Nevertheless, FcR gamma(-/-) mice were highly susceptible to influenza infection, even in the presence of anti-influenza Abs from immune FcR gamma(+/+) mice. NK cells were not necessary for the observed Ab-mediated viral clearance, but macrophages were found to be capable of actively ingesting opsonized virus particles. We conclude that Fc receptor-mediated phagocytosis plays a pivotal role in clearance of respiratory virus infections. PMID- 11390490 TI - Gamma delta T cell-deficient mice have a down-regulated CD8+ T cell immune response against Encephalitozoon cuniculi infection. AB - Gamma(delta) T cells have been reported to play an essential effector role during the early immune response against a wide variety of infectious agents. Recent studies have suggested that the gamma(delta) T cell subtype may also be important for the induction of adaptive immune response against certain microbial pathogens. In the present study, an early increase of gamma(delta) T cells during murine infection with Encephalitozoon cuniculi, an intracellular parasite, was observed. The role of gamma(delta) T cells against E. cuniculi infection was further evaluated by using gene-knockout mice. Mice lacking gamma(delta) T cells were susceptible to E. cuniculi infection at high challenge doses. The reduced resistance of delta(-/-) mice was attributed to a down-regulated CD8+ immune response. Compared with parental wild-type animals, suboptimal Ag-specific CD8+ T cell immunity against E. cuniculi infection was noted in delta(-/-) mice. The splenocytes from infected knockout mice exhibited a lower frequency of Ag specific CD8+ T cells. Moreover, adoptive transfer of immune TCR(alpha)beta+ CD8+ cells from the delta(-/-) mice failed to protect naive CD8(-/-) mice against a lethal E. cuniculi challenge. Our studies suggest that gamma(delta) T cells, due to their ability to produce cytokines, are important for the optimal priming of CD8+ T cell immunity against E. cuniculi infection. This is the first evidence of a parasitic infection in which down-regulation of CD8+ T cell immune response in the absence of gamma(delta) T cells has been demonstrated. PMID- 11390491 TI - Inhibition of Borrelia burgdorferi-tick interactions in vivo by outer surface protein A antibody. AB - Borrelia burgdorferi outer surface protein (Osp) A is preferentially expressed by spirochetes in the Ixodes scapularis gut and facilitates pathogen-vector adherence in vitro. Here we examined B. burgdorferi-tick interactions in vivo by using Abs directed against OspA from each of the three major B. burgdorferi sensu lato genospecies: B. burgdorferi sensu stricto, Borrelia afzelii, and Borrelia garinii. Abs directed against B. burgdorferi sensu stricto (isolate N40) destroy the spirochete and can protect mice from infection. In contrast, antisera raised against OspA from B. afzelii (isolate ACA-1) and B. garinii (isolate ZQ-1) bind to B. burgdorferi N40 but are not borreliacidal against the N40 isolate. Our present studies assess whether these selected OspA Abs interfere with B. burgdorferi-tick attachment in a murine model of Lyme disease with I. scapularis. We examined engorged ticks that had fed on B. burgdorferi N40-infected scid mice previously treated with OspA (N40, ACA-1, ZQ-1, or mAb C3.78) or control Abs. OspA-N40 antisera or mAb C3.78 destroyed B. burgdorferi N40 within the engorged ticks. In contrast, treatment of mice with OspA-ACA-1 and OspA-ZQ-1 antisera did not kill B. burgdorferi N40 within the ticks but did effectively interfere with B. burgdorferi-I. scapularis adherence, thereby preventing efficient colonization of the vector. These studies show that nonborreliacidal OspA Abs can inhibit B. burgdorferi attachment to the tick gut, highlighting the importance of OspA in spirochete-arthropod interactions in vivo. PMID- 11390492 TI - Gestational attenuation of Lyme arthritis is mediated by progesterone and IL-4. AB - Infection of different strains of laboratory mice with the agent of Lyme disease, Borrelia burgdorferi, results in arthritis, the severity of which has been correlated with the dominance of Th1 cytokines. In this study, we demonstrate that changes in B. burgdorferi-specific immunologic responses associated with pregnancy can alter the outcome of Lyme arthritis in mice. Whereas nonpregnant female C3H mice consistently developed severe Lyme arthritis, pregnant mice had a marked reduction in arthritis severity that was associated with a slight reduction in IFN-gamma and markedly increased levels of IL-4 production by B. burgdorferi-specific T cells. Similar reductions in arthritis severity and patterns of cytokine production were observed in nonpregnant, progesterone implanted mice. Ab neutralization of IL-4 in progesterone-implanted mice resulted in severe arthritis. Our results are consistent with the known shift toward Th2 cytokine expression at the maternal-fetal interface, and are the first to show a pregnancy-related therapeutic effect in an infectious model. PMID- 11390493 TI - Transgenic cystic fibrosis mice exhibit reduced early clearance of Pseudomonas aeruginosa from the respiratory tract. AB - The cystic fibrosis (CF) transmembrane conductance regulator (CFTR) has been proposed to be an epithelial cell receptor for Pseudomonas aeruginosa involved in bacterial internalization and clearance from the lung. We evaluated the role of CFTR in clearing P. aeruginosa from the respiratory tract using transgenic CF mice that carried either the DeltaF508 Cftr allele or an allele with a Cftr stop codon (S489X). Intranasal application achieved P. aeruginosa lung infection in inbred C57BL/6 DeltaF508 Cftr mice, whereas DeltaF508 Cftr and S489X Cftr outbred mice required tracheal application of the inoculum to establish lung infection. CF mice showed significantly less ingestion of LPS-smooth P. aeruginosa by lung cells and significantly greater bacterial lung burdens 4.5 h postinfection than C57BL/6 wild-type mice. Microscopy of infected mouse and rhesus monkey tracheas clearly demonstrated ingestion of P. aeruginosa by epithelial cells in wild-type animals, mostly around injured areas of the epithelium. Desquamating cells loaded with P. aeruginosa could also be seen in these tissues. No difference was found between CF and wild-type mice challenged with an LPS-rough mucoid isolate of P. aeruginosa lacking the CFTR ligand. Thus, transgenic CF mice exhibit decreased clearance of P. aeruginosa and increased bacterial burdens in the lung, substantiating a key role for CFTR-mediated bacterial ingestion in lung clearance of P. aeruginosa. PMID- 11390494 TI - Suppression of immune response and protective immunity to a Japanese encephalitis virus DNA vaccine by coadministration of an IL-12-expressing plasmid. AB - IL-12 plays a central role in both innate and acquired immunity and has been demonstrated to potentiate the protective immunity in several experimental vaccines. However, in this study, we show that IL-12 can be detrimental to the immune responses elicited by a plasmid DNA vaccine. Coadministration of the IL-12 expressing plasmid (pIL-12) significantly suppressed the protective immunity elicited by a plasmid DNA vaccine (pE) encoding the envelope protein of Japanese encephalitis virus. This suppressive effect was associated with marked reduction of specific T cell proliferation and Ab responses. A single dose of pIL-12 treatment with plasmid pE in initial priming resulted in significant immune suppression to subsequent pE booster immunization. The pIL-12-mediated immune suppression was dose dependent and evident only when the IL-12 gene was injected either before or coincident with the pE DNA vaccine. Finally, using IFN-gamma gene-disrupted mice, we showed that the suppressive activity of the IL-12 plasmid was dependent upon endogenous production of IFN-gamma. These results demonstrate that coexpression of the IL-12 gene can sometimes produce untoward effects to immune responses, and thus its application as a vaccine adjuvant should be carefully evaluated. PMID- 11390495 TI - Transmission intensity determines lymphocyte responsiveness and cytokine bias in human lymphatic filariasis. AB - Humans living in areas where filariasis is endemic vary greatly in their exposure to mosquito-borne infective third-stage larvae (L3) of these parasitic helminths. Because the intensity of exposure to Ags affects T cell differentiation and susceptibility to parasitic infections in murine models, we compared T cell and cytokine responses in 97 residents of two villages in Papua New Guinea, where transmission intensity of Wuchereria bancrofti differed by 63-fold (37 vs 2355 L3 per person per year). Residents of the high transmission village had 4- to 11 fold lower proliferation and IFN-gamma responses to filarial Ags, nonparasite Ag, and PHA by PBMC compared with the low transmission village (p < 0.01) even when subjects were matched for intensity of infection. In contrast, filarial Ag-driven IL-5 production was 5.5-fold greater (p < 0.001), and plasma IL-4 and TGF-beta levels were 4-fold and 34% higher, respectively, in residents of the high transmission village. IL-4 and IL-10 responses by PBMC differed little according to village, and increased production of the counterregulatory cytokines IL-10 or TGF-beta by PBMC did not correlate with weak proliferation and IFN-gamma responses. Plasma IL-5, IFN-gamma, and IL-10 levels were similar in the two villages. These data demonstrate that the intensity of exposure to L3 affects lymphocyte responsiveness and cytokine bias possibly by a mechanism that alters APC function. PMID- 11390496 TI - Heterosubtypic immunity to influenza A virus in mice lacking IgA, all Ig, NKT cells, or gamma delta T cells. AB - The mechanisms of broad cross-protection to influenza viruses of different subtypes, termed heterosubtypic immunity, remain incompletely understood. We used knockout mouse strains to examine the potential for heterosubtypic immunity in mice lacking IgA, all Ig and B cells, NKT cells (CD1 knockout mice), or gamma(delta) T cells. Mice were immunized with live influenza A virus and compared with controls immunized with unrelated influenza B virus. IgA(-/-) mice survived full respiratory tract challenge with heterosubtypic virus that was lethal to controls. IgA(-/-) mice also cleared virus from the nasopharynx and lungs following heterosubtypic challenge limited to the upper respiratory tract, where IgA has been shown to play an important role. Ig(-/-) mice controlled the replication of heterosubtypic challenge virus in the lungs. Acute depletion of CD4+ or CD8+ T cell subsets abrogated this clearance of virus, thus indicating that both CD4+ and CD8+ T cells are required for protection in the absence of Ig. These results in Ig(-/-) mice indicate that CD4+ T cells can function by mechanisms other than providing help to B cells for the generation of Abs. Like wild-type mice, CD1(-/-) mice and gamma(delta) (-/-) mice survived lethal heterosubtypic challenge. Acute depletion of CD4+ and CD8+ cells abrogated heterosubtypic protection in gamma(delta) (-/-) mice, but not B6 controls, suggesting a contribution of gamma(delta) T cells. Our results demonstrate that the Ab and cellular subsets deficient in these knockout mice are not required for heterosubtypic protection, but each may play a role in a multifaceted response that as a whole is more effective than any of its parts. PMID- 11390497 TI - Immunogenicity of the extracellular domains of C-C chemokine receptor 5 and the in vitro effects on simian immunodeficiency virus or HIV infectivity. AB - The C-C chemokine receptor CCR5 serves an important function in chemotaxis of lymphocytes, monocytes, and dendritic cells. CCR5 is also the major coreceptor in most macrophage-tropic HIV-1 infections. Immunization of rhesus macaques with a baculovirus-generated CCR5 construct or peptides derived from the sequences of the four extracellular domains of CCR5 elicited IgG and IgA Abs, inhibition of SIV replication, and CD4+ T cell proliferative responses to three of the extracellular domains of CCR5. The immune sera reacted with cell surface CCR5 expressed on HEK 293 cells. T and B cell epitope mapping revealed major and minor T and B cell epitopes in the N-terminal, first, and second loops of CCR5. The three C-C chemokines, RANTES, macrophage-inflammatory protein-1alpha, and macrophage-inflammatory protein-1beta, were up-regulated by immunization with the CCR5-derived peptides, and the cell surface expression of CCR5 was decreased. The CCR5 Abs were complementary to the C-C chemokines in inhibiting HIV replication in vitro. Immunization with the four extracellular domains of CCR5 suggests that three of them are immunogenic, with maximal T cell responses being elicited by the second loop peptide. However, maximal Abs to the cell surface CCR5 or viral inhibitory Abs in vitro were induced by the N-terminal peptide. Up-regulation of the three C-C chemokines and down-modulation of cell surface CCR5 were elicited by the second loop, N-terminal, and first loop peptides. The data suggest that a dual mechanism of C-C chemokines and specific Abs may engage and down-modulate the CCR5 coreceptors and prevent in vitro HIV or SIV replication. PMID- 11390498 TI - The role of T cell subsets and cytokines in the pathogenesis of Helicobacter pylori gastritis in mice. AB - Gastritis due to Helicobacter pylori in mice and humans is considered a Th1 mediated disease, but the specific cell subsets and cytokines involved are still not well understood. The goal of this study was to investigate the immunopathogenesis of H. pylori-induced gastritis and delayed-type hypersensitivity (DTH) in mice. C57BL/6-Prkdc(scid) mice were infected with H. pylori and reconstituted with CD4+, CD4-depleted, CD4+CD45RB(high), or CD4+CD45RB(low) splenocytes from wild-type C57BL/6 mice or with splenocytes from C57BL/6(IFN-gamma-/-) or C57BL/6(IL-10-/-) mice. Four or eight weeks after transfer, DTH to H. pylori Ags was determined by footpad injection; gastritis and bacterial colonization were quantified; and IFN-gamma secretion by splenocytes in response to H. pylori Ag was determined. Gastritis and DTH were present in recipients of unfractionated splenocytes, CD4+ splenocytes, and CD4+CD45RB(high) splenocytes, but absent in the other groups. IFN-gamma secretion in response to H. pylori Ags was correlated with gastritis, although splenocytes from all groups of mice secreted some IFN-gamma. Gastritis was most severe in recipients of splenocytes from IL-10-deficient mice, and least severe in those given IFN-gamma deficient splenocytes. Bacterial colonization in all groups was inversely correlated with gastritis. These data indicate that 1) CD4+ T cells are both necessary and sufficient for gastritis and DTH due to H. pylori in mice; 2) high expression of CD45RB is a marker for gastritis-inducing CD4+ cells; and 3) IFN gamma contributes to gastritis and IL-10 suppresses it, but IFN-gamma secretion alone is not sufficient to induce gastritis. The results support the assertion that H. pylori is mediated by a Th1-biased cellular immune response. PMID- 11390499 TI - A filarial nematode-secreted phosphorylcholine-containing glycoprotein uncouples the B cell antigen receptor from extracellular signal-regulated kinase-mitogen activated protein kinase by promoting the surface Ig-mediated recruitment of Src homology 2 domain-containing tyrosine phosphatase-1 and Pac-1 mitogen-activated kinase-phosphatase. AB - Unraveling the molecular mechanisms by which filarial nematodes, major human pathogens in the tropics, evade the host immune system remains an elusive goal. We have previously shown that excretory-secretory product-62 (ES-62), a homologue of phosphorylcholine-containing molecules that are secreted by human parasites and which is active in rodent models of filarial infection, is able to polyclonally activate certain protein tyrosine kinase and mitogen-activating protein kinase signal transduction elements in B lymphocytes. Such activation mediates desensitization of subsequent B cell Ag receptor (BCR) ligation-induced activation of extracellular signal-regulated kinase-mitogen-activated protein (ErkMAP) kinase and ultimately B cell proliferation. We now show that the desensitization is due to ES-62 targeting two major regulatory sites of B cell activation. Firstly, pre-exposure to ES-62 primes subsequent BCR-mediated recruitment of SHP-1 tyrosine phosphatase to abolish recruitment of the RasErkMAP kinase cascade via the Igalphabeta-ShcGrb2Sos adaptor complex interactions. Secondly, any ongoing ErkMAP kinase signaling in ES-62-primed B cells is terminated by the MAP kinase phosphatase, Pac-1 that is activated consequently to challenge via the BCR. PMID- 11390500 TI - Cytosolic phospholipase A2 participates with TNF-alpha in the induction of apoptosis of human macrophages infected with Mycobacterium tuberculosis H37Ra. AB - Macrophage (MPhi) apoptosis, an important innate microbial defense mechanism induced by Mycobacterium tuberculosis (Mtb) H37Ra, depends on the induction of TNF-alpha synthesis. When protein synthesis is blocked, both infection with Mtb and addition of TNF-alpha are required to induce caspase 9 activation, caspase 3 activation and apoptosis. In this study, we show that the second protein synthesis-independent signal involves activation of group IV cytosolic phospholipase A2 (cPLA2). Apoptosis of Mtb-infected MPhi and concomitant arachidonic acid release are abrogated by group IV cPLA2 inhibitors (methyl arachidonyl fluorophosphate and methyl trifluoromethyl ketone), but not by inhibitors of group VI Ca2+-independent (iPLA2; bromoenol lactone) or of secretory low molecular mass PLA2. In MPhi homogenates, the predominant PLA2 activity showed the same inhibitor sensitivity pattern and preferred arachidonic acid over palmitic acid in substrates, also indicating the presence of one or more group IV cPLA2 enzymes. In concordance with these findings, MPhi lysates contained transcripts and protein for group IV cPLA2-alpha and cPLA2-gamma. Importantly, group IV cPLA2 inhibitors significantly reduced MPhi antimycobacterial activity and addition of arachidonic acid, the major product of group IV cPLA2, to infected MPhi treated with cPLA2 inhibitors completely restored the antimycobacterial activity. Importantly, addition of arachidonic acid alone to infected MPhi significantly reduced the mycobacterial burden. These findings indicate that Mtb induces MPhi apoptosis by independent signaling through at least two pathways, TNF-alpha and cPLA2, which are both also critical for antimycobacterial defense of the MPhi. PMID- 11390501 TI - Mannosylated lipoarabinomannans inhibit IL-12 production by human dendritic cells: evidence for a negative signal delivered through the mannose receptor. AB - IL-12 is a key cytokine in directing the development of type 1 Th cells, which are critical to eradicate intracellular pathogens such as Mycobacterium tuberculosis. Here, we report that mannose-capped lipoarabinomannans (ManLAMs) from Mycobacterium bovis bacillus Calmette-Guerin and Mycobacterium tuberculosis inhibited, in a dose-dependent manner, the LPS-induced IL-12 production by human dendritic cells. The inhibitory activity was abolished by the loss of the mannose caps or the GPI acyl residues. Mannan, which is a ligand for the mannose receptor (MR) as well as an mAb specific for the MR, also inhibited the LPS-induced IL-12 production by dendritic cells. Our results indicate that ManLAMs may act as virulence factors that contribute to the persistence of M. bovis bacillus Calmette-Guerin and M. tuberculosis within phagocytic cells by suppressing IL-12 responses. Our data also suggest that engagement of the MR by ManLAMs delivers a negative signal that interferes with the LPS-induced positive signals delivered by the Toll-like receptors. PMID- 11390502 TI - Cooperative regulation of Mcl-1 by Janus kinase/stat and phosphatidylinositol 3 kinase contribute to granulocyte-macrophage colony-stimulating factor-delayed apoptosis in human neutrophils. AB - Polymorphonuclear neutrophils (PMN) are phagocytic cells constitutively programmed for apoptotic cell death. Exposure to GM-CSF delays apoptosis as measured by annexin-V staining and cell morphological change. We found that STAT5B, STAT1, and STAT3 DNA-binding activity was induced by GM-CSF. We also detected activation of the phosphatidylinositol 3-kinase (PI 3-kinase) pathway after GM-CSF treatment which was inhibited by treatment with the PI 3-kinase inhibitors, wortmannin and LY294002. We investigated whether STAT or PI 3-kinase activity was necessary for the pro-survival response of GM-CSF in PMN. Exposure of PMN to GM-CSF in the presence of either AG-490, antisense STAT3 oligonucleotides, or wortmannin resulted in a partial inhibition of GM-CSF mediated pro-survival activity. GM-CSF induced a time-dependent increase in the mRNA and protein expression of the anti-apoptotic Bcl-2-family protein, Mcl-1. We examined the hypothesis that Janus kinase/STAT and PI 3-kinase regulation of Mcl 1 contributed to GM-CSF-delayed apoptosis. Using either AG-490 or wortmannin alone, we observed a dose-dependent inhibition of GM-CSF-induced Mcl-1 expression. Using suboptimal doses of AG-490 and wortmannin, we found that both drugs together had an additive effect on delayed apoptosis and Mcl-1 expression. These data suggest that cooperative regulation of Mcl-1 by the Janus kinase/STAT and PI 3-kinase pathways contribute to GM-CSF-delayed apoptosis. PMID- 11390503 TI - Antibody-mediated phagocytosis of the amyloid beta-peptide in microglia is differentially modulated by C1q. AB - Microglial ingestion of the amyloid beta-peptide (Abeta) has been viewed as a therapeutic target in Alzheimer's disease, in that approaches that enhance clearance of Abeta relative to its production are predicted to result in decreased senile plaque formation, a proposed contributor to neuropathology. In vitro, scavenger receptors mediate ingestion of fibrillar Abeta (fAbeta) by microglia. However, the finding that cerebral amyloid deposition in a transgenic mouse model of Alzheimer's disease was diminished by inoculation with synthetic Abeta has suggested a possible therapeutic role for anti-Abeta Ab-mediated phagocytosis. Microglia also express C1qR(P), a receptor for complement protein C1q, ligation of which in vitro enhances phagocytosis of immune complexes formed with IgG levels below that required for optimal FcR-mediated phagocytosis. The data presented here demonstrate FcR-dependent ingestion of Abeta-anti-Abeta complexes (IgG-fAbeta) by microglia that is a function of the amount of Ab used to form immune complexes. In addition, C1q incorporated into IgG-fAbeta enhanced microglial uptake of these complexes when they contained suboptimal levels of anti-Abeta Ab. Mannose binding lectin and lung surfactant protein A, other ligands of C1qR(P), also enhanced ingestion of suboptimally opsonized IgG-fAbeta, whereas control proteins did not. Our data suggest that C1qR(P)-mediated events may promote efficient ingestion of Abeta at low Ab titers, and this may be beneficial in paradigms that seek to clear amyloid via FcR-mediated mechanisms by minimizing the potential for destructive Ab-induced complement-mediated processes. PMID- 11390504 TI - IL-12 suppression during experimental endotoxin tolerance: dendritic cell loss and macrophage hyporesponsiveness. AB - Endotoxin tolerance, the transient, secondary down-regulation of a subset of endotoxin-driven responses after exposure to bacterial products, is thought to be an adaptive response providing protection from pathological hyperactivation of the innate immune system during bacterial infection. However, although protecting from the development of sepsis, endotoxin tolerance also can lead to fatal blunting of immunological responses to subsequent infections in survivors of septic shock. Despite considerable experimental effort aimed at characterizing the molecular mechanisms responsible for a variety of endotoxin tolerance-related phenomena, no consensus has been achieved yet. IL-12 is a macrophage- and dendritic cell (DC)-derived cytokine that plays a key role in pathological responses to endotoxin as well as in the induction of protective responses to pathogens. It recently has been shown that IL-12 production is suppressed in endotoxin tolerance, providing a likely partial mechanism for the increased risk of secondary infections in sepsis survivors. We examined the development of IL-12 suppression during endotoxin tolerance in mice. Decreased IL-12 production in vivo is clearly multifactorial, involving both loss of CD11c(high) DCs as well as alterations in the responsiveness of macrophages and remaining splenic DCs. We find no demonstrable mechanistic role for B or T lymphocytes, the soluble mediators IL-10, TNF-alpha, IFN-alphabeta, or nitric oxide, or the NF-kappaB family members p50, p52, or RelB. PMID- 11390505 TI - Surfactant protein D regulates NF-kappa B and matrix metalloproteinase production in alveolar macrophages via oxidant-sensitive pathways. AB - Targeted ablation of the surfactant protein D (SP-D) gene caused progressive pulmonary emphysema associated with pulmonary infiltration by foamy alveolar macrophages (AMs), increased hydrogen peroxide production, and matrix metalloproteinase (MMP)-2, -9, and -12 expression. In the present study, the mechanisms by which SP-D influences macrophage MMP activity were assessed in AMs from SP-D(-/-) mice. Tissue lipid peroxides and reactive carbonyls were increased in lungs of SP-D(-/-) mice, indicating oxidative stress. Immunohistochemical staining of AMs from SP-D(-/-) mice demonstrated that NF-kappaB was highly expressed and translocated to the nucleus. Increased NF-kappaB binding was detected by EMSA in nuclear extracts of AMs isolated from SP-D(-/-) mice. Antioxidants N-acetylcysteine and pyrrolidine dithiocarbamate inhibited MMP production by AMs from SP-D(-/-) mice. To assess whether increased oxidant production influenced NF-kappaB activation and production of MMP-2 and -9, AMs from SP-D(-/-) mice were treated with the NADPH oxidase inhibitors diphenylene iodonium chloride and apocynin. Inhibition of NADPH oxidase suppressed NF-kappaB binding by nuclear extracts and decreased production of MMP-2 and 9 in AMs from SP-D(-/-) mice. SN-50, a synthetic NF-kappaB-inhibitory peptide, decreased MMP production by AMs from SP-D(-/-) mice. Oxidant production and reactive oxygen species were increased in lungs of SP-D(-/-) mice, in turn activating NF-kappaB and MMP expression. SP-D plays an unexpected inhibitory role in the regulation of NF-kappaB in AMs. PMID- 11390507 TI - Minocycline provides neuroprotection against N-methyl-D-aspartate neurotoxicity by inhibiting microglia. AB - Glutamate excitotoxicity to a large extent is mediated through activation of the N-methyl-D-aspartate (NMDA)-gated ion channels in several neurodegenerative diseases and ischemic stroke. Minocycline, a tetracycline derivative with antiinflammatory effects, inhibits IL-1beta-converting enzyme and inducible nitric oxide synthase up-regulation in animal models of ischemic stroke and Huntington's disease and is therapeutic in these disease animal models. Here we report that nanomolar concentrations of minocycline protect neurons in mixed spinal cord cultures against NMDA excitotoxicity. NMDA treatment alone induced microglial proliferation, which preceded neuronal death, and administration of extra microglial cells on top of these cultures enhanced the NMDA neurotoxicity. Minocycline inhibited all these responses to NMDA. Minocycline also prevented the NMDA-induced proliferation of microglial cells and the increased release of IL 1beta and nitric oxide in pure microglia cultures. Finally, minocycline inhibited the NMDA-induced activation of p38 mitogen-activated protein kinase (MAPK) in microglial cells, and a specific p38 MAPK inhibitor, but not a p44/42 MAPK inhibitor, reduced the NMDA toxicity. Together, these results suggest that microglial activation contributes to NMDA excitotoxicity and that minocycline, a tetracycline derivative, represents a potential therapeutic agent for brain diseases. PMID- 11390506 TI - A CD2-green fluorescence protein-transgenic mouse reveals very late antigen-4 dependent CD8+ lymphocyte rolling in inflamed venules. AB - Intravital microscopy allows detailed analysis of leukocyte trafficking in vivo, but fails to identify the nature of leukocytes investigated. Here, we describe the development of a CD2-enhanced green fluorescence protein (EGFP)-transgenic mouse to characterize lymphocyte trafficking during inflammation in vivo. A CD2 EGFP plasmid construct including the CD2 promoter, the EGFP transgene, and the CD2 locus control region was injected into B6CBA/F1 pronuclei. EGFP+ offspring were backcrossed into C57BL/6 mice for six generations. Flow cytometry demonstrated that all peripheral blood EGFP+ cells were positive for CD2 and negative for the granulocyte Ag Ly 6-G (GR-1). EGFP(high) cells stained positive for CD2, CD3, CD8, TCR beta-chain, and NK1.1 but did not express the B cell and monocyte markers CD45RA, CD19, and CD11b. In vitro stimulation assays revealed no difference in lymphocyte proliferation and IL-2 secretion between EGFP+ and EGFP- mice. Intravital microscopy of untreated or TNF-alpha-treated cremaster muscle venules showed EGFP+ cells in vivo, but these cells did not roll or adhere to the vessel wall. In cremaster muscle venules treated with both TNF-alpha and IFN gamma, EGFP(high) cells rolled, adhered, and transmigrated at a rolling velocity slightly higher (11 microm/s) than that of neutrophils (10 microm/s). Blocking alpha4 integrin with a mAb increased rolling velocity to 24 microm/s. These findings show that CD8+ T cells roll in TNF-alpha/IFN-gamma-pretreated vessels in vivo via an alpha4 integrin-dependent pathway. PMID- 11390508 TI - Blockade of alpha 5 beta 1 integrins reverses the inhibitory effect of tenascin on chemotaxis of human monocytes and polymorphonuclear leukocytes through three dimensional gels of extracellular matrix proteins. AB - Tenascin is an extracellular matrix protein found in adults in T cell-dependent areas of lymphoid tissues, sites of inflammation, and tumors. We report here that it inhibited chemotaxis of chemoattractant-stimulated human monocytes and chemoattractant-stimulated polymorphonuclear leukocytes (PMN) through three dimensional gels composed of collagen I or Matrigel, and chemotaxis of leukotriene B4-stimulated PMN through fibrin gels. The inhibitory effect of tenascin on monocyte or PMN chemotaxis through these matrices was reversed by Abs directed against alpha5beta1 integrins or by a peptide (GRGDSP) that binds to beta1 integrins. Tenascin did not affect leukotriene B4- or fMLP-stimulated expression of beta1 or beta2 integrins, but did exert a small inhibitory effect on PMN adhesion and closeness of apposition to fibrin(ogen)-containing surfaces. Thus, alpha5beta1 integrins mediate the inhibitory effect of tenascin on monocyte and PMN chemotaxis, without promoting close apposition between these leukocytes and surfaces coated with tenascin alone or with tenascin bound to other matrix proteins. This contrasts with the role played by alpha5beta1 integrins in promoting close apposition between fMLP-stimulated PMN and fibrin containing surfaces, thereby inhibiting chemotaxis of fMLP-stimulated PMN through fibrin gels. Thus, chemoattractants and matrix proteins regulate chemotaxis of phagocytic leukocytes by at least two different mechanisms: one in which specific chemoattractants promote very tight adhesion of leukocytes to specific matrix proteins and another in which specific matrix proteins signal cessation of migration without markedly affecting strength of leukocyte adhesion. PMID- 11390509 TI - Hypoxia-inducible factor 1-mediated inhibition of peroxisome proliferator activated receptor alpha expression during hypoxia. AB - Peroxisome proliferator-activated receptors (PPARs) are nuclear hormone-binding proteins that regulate transcriptional responses to peroxisome proliferators and structurally diverse fatty acids. PPARs have been implicated in a wide variety of functions, including lipid homeostasis and inflammatory responses. In this study, we examined the expression of PPAR-alpha in response to ambient hypoxia. Initial studies using microarray analysis of intestinal epithelial mRNA revealed that hypoxia rapidly down-regulates PPAR-alpha mRNA and protein in epithelial cells in vitro and in vivo. Subsequent studies revealed that the PPAR-alpha gene bears a DNA consensus motif for the transcription factor hypoxia-inducible factor 1 (HIF 1). EMSA analysis revealed that ambient hypoxia induces HIF-1alpha binding to the HIF-1 consensus domain of PPAR-alpha in parallel to HIF-1 nuclear accumulation, and antisense depletion of HIF-1alpha resulted in a loss of PPAR-alpha down regulation. The PPAR-alpha ligand pirinixic acid (WY14643) functionally promoted IFN-gamma-induced ICAM-1 expression in normoxic epithelia, and this response was lost in cells pre-exposed to ambient hypoxia. Such results indicate that HIF-1 dependent down-regulation of PPAR-alpha may provide an adaptive response to proinflammatory stimuli during cellular hypoxia. These studies provide unique insight into the regulation of PPAR-alpha expression and, importantly, provide an example of a down-regulatory pathway mediated by HIF-1. PMID- 11390510 TI - Non-small cell lung cancer cells induce monocytes to increase expression of angiogenic activity. AB - Tumors are dependent on angiogenesis for survival and propagation. Accumulated evidence suggests that macrophages are a potentially important source of angiogenic factors in many disease states. However, the role(s) of macrophages in non-small cell lung cancer (NSCLC) have not been determined. We hypothesized that monocyte-derived macrophages are induced by NSCLC to increase expression of angiogenic factors. To define the role of macrophage-tumor cell interaction with respect to angiogenesis, human peripheral blood monocytes (PBM) were cocultured with A549 (human bronchoalveolar cell carcinoma) or Calu 6 (human anaplastic carcinoma) NSCLC cells. The resultant conditioned medium (CM) was evaluated for angiogenic potential and for expression of angiogenic factors. We found that endothelial cell chemotactic activity (as a measure of angiogenic potential) was significantly increased in response to CM from cocultures of PBM/NSCLC compared with PBM alone, NSCLC alone, or a combination of NSCLC and PBM CM generated separately. Subsequent analysis by ELISA reveals markedly increased CXC chemokine expression, with a lesser increase in vascular endothelial growth factor, in CM from PBM/NSCLC coculture. Neutralizing Ab to angiogenic CXC chemokines blocked the increase in endothelial cell chemotaxis. Furthermore, with separately generated CM as a stimulus, we found that macrophages are the predominant source of increased CXC chemokine expression. Finally, we found that NSCLC-derived macrophage migration-inhibitory factor is responsible for the increased expression of macrophage-derived angiogenic activity. These data suggest that the interaction between host macrophages and NSCLC cells synergistically increases angiogenic potential, and that this is due to an increased elaboration of angiogenic CXC chemokines. PMID- 11390511 TI - Peripheral blood fibrocytes: differentiation pathway and migration to wound sites. AB - Fibrocytes are a distinct population of blood-borne cells that display a unique cell surface phenotype (collagen I+/CD11b+/CD13+/CD34+/CD45RO+/MHC class II+/CD86+) and exhibit potent immunostimulatory activities. Circulating fibrocytes rapidly enter sites of tissue injury, suggesting an important role for these cells in wound repair. However, the regulatory processes that govern the differentiation of blood-borne fibrocytes and the mechanisms that underlie the migration of these cells to wound sites are currently not known. We report herein that ex vivo cultured fibrocytes can differentiate from a CD14+-enriched mononuclear cell population and that this process requires contact with T cells. Furthermore, we demonstrate that TGF-beta1 (1-10 ng/ml), an important fibrogenic and growth-regulating cytokine involved in wound healing, increases the differentiation and functional activity of cultured fibrocytes. Because fibrocytes home to sites of tissue injury, we examined the role of chemokine/chemokine receptor interactions in fibrocyte trafficking. We show that secondary lymphoid chemokine, a ligand of the CCR7 chemokine receptor, acts as a potent stimulus for fibrocyte chemotaxis in vitro and for the homing of injected fibrocytes to sites of cutaneous tissue injury in vivo. Finally, we demonstrate that differentiated, cultured fibrocytes express alpha smooth muscle actin and contract collagen gels in vitro, two characteristic features of wound-healing myofibroblasts. These data provide important insight into the control of fibrocyte differentiation and trafficking during tissue repair and significantly expand their potential role during wound healing. PMID- 11390512 TI - Ubiquitous transgenic expression of the IL-23 subunit p19 induces multiorgan inflammation, runting, infertility, and premature death. AB - p19, a molecule structurally related to IL-6, G-CSF, and the p35 subunit of IL 12, is a subunit of the recently discovered cytokine IL-23. Here we show that expression of p19 in multiple tissues of transgenic mice induced a striking phenotype characterized by runting, systemic inflammation, infertility, and death before 3 mo of age. Founder animals had infiltrates of lymphocytes and macrophages in skin, lung, liver, pancreas, and the digestive tract and were anemic. The serum concentrations of the proinflammatory cytokines TNF-alpha and IL-1 were elevated, and the number of circulating neutrophils was increased. In addition, ubiquitous expression of p19 resulted in constitutive expression of acute phase proteins in the liver. Surprisingly, liver-specific expression of p19 failed to reproduce any of these abnormalities, suggesting specific requirements for production of biologically active p19. Bone marrow transfer experiments showed that expression of p19 by hemopoietic cells alone recapitulated the phenotype induced by its widespread expression, pointing to hemopoietic cells as the source of biologically active p19. These findings indicate that p19 shares biological properties with IL-6, IL-12, and G-CSF and that cell-specific expression is required for its biological activity. PMID- 11390513 TI - Eotaxin (CCL11) induces in vivo angiogenic responses by human CCR3+ endothelial cells. AB - Chemokines are attractants and regulators of cell activation. Several CXC family chemokine members induce angiogenesis and promote tumor growth. In contrast, the only CC chemokine, reported to play a direct role in angiogenesis is monocyte chemotactic protein-1. Here we report that another CC chemokine, eotaxin (also known as CCL11), also induced chemotaxis of human microvascular endothelial cells. CCL11-induced chemotactic responses were comparable with those induced by monocyte-chemotactic protein-1 (CCL2), but lower than those induced by stroma derived factor-1alpha (CXCL12) and IL-8 (CXCL8). The chemotactic activity was consistent with the expression of CCR3, the receptor for CCL11, on human microvascular endothelial cells and was inhibited by mAbs to either human CCL11 or human CCR3. CCL11 also induced the formation of blood vessels in vivo as assessed by the chick chorioallantoic membrane and Matrigel plug assays. The angiogenic response induced by CCL11 was about one-half of that induced by basic fibroblast factor, and it was accompanied by an inflammatory infiltrate, which consisted predominantly of eosinophils. Because the rat aortic sprouting assay, which is not infiltrated by eosinophils, yielded a positive response to CCL11, this angiogenic response appears to be direct and is not mediated by eosinophil products. This suggests that CCL11 may contribute to angiogenesis in conditions characterized by increased CCL11 production and eosinophil infiltration such as Hodgkin's lymphoma, nasal polyposis, endometriosis, and allergic diathesis. PMID- 11390514 TI - Myelin antigen-specific CD8+ T cells are encephalitogenic and produce severe disease in C57BL/6 mice. AB - Encephalitogenic T cells that mediate experimental autoimmune encephalomyelitis (EAE) are commonly assumed to be exclusively CD4+, but formal proof is still lacking. In this study, we report that synthetic peptides 35-55 from myelin oligodendrocyte glycoprotein (pMOG(35-55)) consistently activate a high proportion of CD8+ alphabetaTCR+ T cells that are encephalitogenic in C57BL/6 (B6) mice. The encephalitogenic potential of CD8+ MOG-specific T cells was established by adoptive transfer of CD8-enriched MOG-specific T cells. These cells induced a much more severe and permanent disease than disease actively induced by immunization with pMOG(35-55). CNS lesions in pMOG(35-55) CD8+ T cell induced EAE were progressive and more destructive. The CD8+ T cells were strongly pathogenic in syngeneic B6 and RAG-1(-/-) mice, but not in isogeneic beta2 microglobulin-deficient mice. MOG-specific CD8+ T cells could be repeatedly reisolated for up to 287 days from recipient B6 or RAG-1(-/-) mice in which disease was induced adoptively with <1 x 10(6) T cells sensitized to pMOG(35-55). It is postulated that MOG induces a relapsing and/or progressive pattern of EAE by eliciting a T cell response dominated by CD8+ autoreactive T cells. Such cells appear to have an enhanced tissue-damaging effect and persist in the animal for long periods. PMID- 11390515 TI - MHC class II-regulated central nervous system autoaggression and T cell responses in peripheral lymphoid tissues are dissociated in myelin oligodendrocyte glycoprotein-induced experimental autoimmune encephalomyelitis. AB - We dissected the requirements for disease induction of myelin oligodendrocyte glycoprotein (MOG)-induced experimental autoimmune encephalomyelitis in MHC (RT1 in rat) congenic rats with overlapping MOG peptides. Immunodominance with regard to peptide-specific T cell responses was purely MHC class II dependent, varied between different MHC haplotypes, and was linked to encephalitogenicity only in RT1.B(a)/D(a) rats. Peptides derived from the MOG sequence 91-114 were able to induce overt clinical signs of disease accompanied by demyelinated CNS lesions in the RT1.B(a)/D(a) and RT1(n) haplotypes. Notably, there was no detectable T cell response against this encephalitogenic MOG sequence in the RT1(n) haplotype in peripheral lymphoid tissue. However, CNS-infiltrating lymphoid cells displayed high IFN-gamma, TNF-alpha, and IL-4 mRNA expression suggesting a localization of peptide-specific reactivated T cells in this compartment. Despite the presence of MOG-specific T and B cell responses, no disease could be induced in resistant RT1(l) and RT1(u) haplotypes. Comparison of the number of different MOG peptides binding to MHC class II molecules from the different RT1 haplotypes suggested that susceptibility to MOG-experimental autoimmune encephalomyelitis correlated with promiscuous peptide binding to RT1.B and RT1.D molecules. This may suggest possibilities for a broader repertoire of peptide-specific T cells to participate in disease induction. We demonstrate a powerful MHC class II regulation of autoaggression in which MHC class II peptide binding and peripheral T cell immunodominance fail to predict autoantigenic peptides relevant for an autoaggressive response. Instead, target organ responses may be decisive and should be further explored. PMID- 11390516 TI - Protein kinase A RI beta subunit deficiency in lupus T lymphocytes: bypassing a block in RI beta translation reconstitutes protein kinase A activity and augments IL-2 production. AB - A profound deficiency of type I protein kinase A (PKA-I or RIalpha/beta2C2) phosphotransferase activity occurs in the T lymphocytes of 80% of subjects with systemic lupus erythematosus (SLE), an autoimmune disorder of unknown etiology. This isozyme deficiency is predominantly the product of reduced or absent beta isoform of the type I regulatory subunit (RIbeta). Transient transfection of RIbeta cDNAs from SLE subjects into autologous T cells that do not synthesize the RIbeta subunit bypassed the block, resulting in RIbeta subunit synthesis and restoration of the PKA-Ibeta (RIbeta2C2) holoenzyme. Transfected T cells activated via the T cell surface receptor complex revealed a significant increase of cAMP-activatable PKA activity that was associated with a significant increase in IL-2 production. These data demonstrate that a disorder of RIbeta translation exists, and that correction of the PKA-I deficiency may enhance T lymphocyte effector functions in SLE. PMID- 11390517 TI - Antibodies to C-C chemokine receptor 5 in normal human IgG block infection of macrophages and lymphocytes with primary R5-tropic strains of HIV-1. AB - In the present study, we demonstrate that normal human IgG for therapeutic use (i.v. Ig) contains natural Abs directed against the CCR5 coreceptor for HIV-1. Abs to CCR5 were isolated from i.v. Ig using an affinity matrix consisting of a synthetic peptide corresponding to the N-terminus of CCR5 coupled to Sepharose. Natural anti-CCR5 Abs inhibited the binding of RANTES to macrophages, demonstrating their interaction with the coreceptor of R5-tropic HIV-1. Affinity purified anti-CCR5 Ig further inhibited infection of lymphocytes and monocytes/macrophages with primary and laboratory-adapted strains of HIV-1, but did not inhibit infection with X4-tropic HIV. Our results suggest that anti-CCR5 Abs from healthy immunocompetent donors may be suitable for development of novel passive immunotherapy regimens in specific clinical settings in HIV infection. PMID- 11390518 TI - Molecular basis of a selective C1s deficiency associated with early onset multiple autoimmune diseases. AB - We have investigated the molecular basis of selective and complete C1s deficiency in 2-year-old girl with complex autoimmune diseases including lupus-like syndrome, Hashimoto's thyroiditis, and autoimmune hepatitis. This patient's complement profile was characterized by the absence of CH50 activity, C1 functional activity <10%, and undetectable levels of C1s Ag associated with normal levels of C1r and C1q Ags. Exon-specific amplification of genomic DNA by PCR followed by direct sequence analysis revealed a homozygous nonsense mutation in the C1s gene exon XII at codon 534, caused by a nucleotide substitution from C (CGA for arginine) to T (TGA for stop codon). Both parents were heterozygous for this mutation. We used the new restriction site for endonuclease Fok-1 created by the mutation to detect this mutation in the genomic DNA of seven healthy family members. Four additional heterozygotes for the mutation were identified in two generations. Our data characterize for the first time the genetic defect of a selective and complete C1s deficiency in a Caucasian patient. PMID- 11390519 TI - CXCL10 (IFN-gamma-inducible protein-10) control of encephalitogenic CD4+ T cell accumulation in the central nervous system during experimental autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is a CD4(+) Th1-mediated demyelinating disease of the CNS that serves as a model for multiple sclerosis. A critical event in the pathogenesis of EAE is the entry of both Ag-specific and Ag nonspecific T lymphocytes into the CNS. In the present report, we investigated the role of the CXC chemokine CXCL10 (IFN-gamma-inducible protein-10) in the pathogenesis of EAE. Production of CXCL10 in the CNS correlated with the development of clinical disease. Administration of anti-CXCL10 decreased clinical and histological disease incidence, severity, as well as infiltration of mononuclear cells into the CNS. Anti-CXCL10 specifically decreased the accumulation of encephalitogenic PLP(139-151) Ag-specific CD4+ T cells in the CNS compared with control-treated animals. Anti-CXCL10 administration did not affect the activation of encephalitogenic T cells as measured by Ag-specific proliferation and the ability to adoptively transfer EAE. These results demonstrate an important role for the CXC chemokine CXCL10 in the recruitment and accumulation of inflammatory mononuclear cells during the pathogenesis of EAE. PMID- 11390520 TI - The prevention and treatment of murine colitis using gene therapy with adenoviral vectors encoding IL-10. AB - IL-10-deficient (IL-10(-/-)) mice develop colitis with many similarities to Crohn's disease. Daily IL-10 injections have a short systemic half-life and are unable to induce complete remission in IL-10(-/-) mice with established disease. In this paper, we investigate the duration, potency, and immunogenicity of gene therapy using an adenoviral vector encoding murine IL-10 (AdvmuIL-10). A single systemic injection of AdvmuIL-10 was sufficient not only to prevent the onset of colitis for at least 10 wk but also to induce clinical and histological remission in mice with established disease. In addition, AdvmuIL-10 diminished the systemic manifestations of disease, including elevated acute-phase proteins, as well as the local consequences of inflammation such as raised stool IL-1beta concentrations. Both IL-10 protein and the effects of secreted IL-10 were detectable for 10 wk after AdvmuIL-10 injection. Furthermore, the immunoregulatory effect of a single AdvmuIL-10 injection was manifest both by a reduction in TNF-alpha, IFN-gamma, and RANTES release from stimulated splenocyte cultures, and also by a change in the proportion of CD45RB(high/low) lymphocytes in the spleen compared with control mice. The delivery of AdvmuIL-10 resulted in a significantly diminished host antiadenoviral response compared with control adenoviral vectors. Thus, gene therapy strategies using adenoviral vectors encoding immunoregulatory and antiinflammatory cytokines may prove to be a potent approach for the treatment of chronic inflammatory disease. Antiinflammatory cytokine expression protects against immune responses directed at gene vectors. PMID- 11390522 TI - Stool DNA-based colorectal cancer detection: finding the needle in the haystack. PMID- 11390521 TI - Ex vivo IFN-gamma secretion by circulating CD8 T lymphocytes: implications of a novel approach for T cell monitoring in infectious and malignant diseases. AB - To elucidate the functional heterogeneity of Ag-specific T lymphocyte populations, we combined labeling of lymphocytes with MHC/peptide tetramers and a cell surface affinity matrix for IFN-gamma. Magnetic cell sorting of IFN-gamma positive lymphocytes allowed the selective enrichment and identification of live Ag-specific cytokine-secreting cells by flow cytometry. Naive, memory, and effector Ag-specific populations were evaluated in healthy HLA-A2 individuals. Significant fractions of influenza- and CMV-specific cells secreted IFN-gamma upon challenge with cognate peptide, consistent with an effector/memory status. The sensitivity of the approach allowed the detection of significant numbers of CMV-specific IFN-gamma-secreting cells ex vivo (i.e., without Ag stimulation). This was not apparent when using previously described assays, namely, ELISPOT or intracellular IFN-gamma staining (cytospot). CD8+ T cells specific for the melamoma-associated Ag Melan-A/MART-1 did not produce IFN-gamma upon challenge with cognate peptide, reminiscent with their naive functional state in healthy individuals. In contrast, CD45RA(low) Melan-A/MART-1 tumor-specific cells from three of three melanoma patients presented levels of activity similar to those found for influenza- or CMV virus-specific lymphocytes, compatible with a functional differentiation into competent effector/memory T lymphocytes in vivo. Notably, a sizable fraction of Melan-A/MART-1-specific cells from a patient secreted IFN-gamma ex vivo following peptide-based vaccination. Thus, the high sensitivity of the assay provides a valuable tool to monitor effector T cell responses in different clinical situations. PMID- 11390523 TI - Telomerase still 'investigational' in its application to cancer management. PMID- 11390524 TI - Studies move closer to genetic test to detect colon cancer. PMID- 11390525 TI - Van Andel Institute brings cancer research to the heartland. PMID- 11390526 TI - Medicare coverage group assesses effectiveness of new technologies. PMID- 11390528 TI - Does type of health insurance affect cancer care, diagnosis? PMID- 11390531 TI - Behavioral intervention for cancer treatment side effects. AB - The use of increasingly aggressive methods of cancer treatment during the last 20 years has brought clinical attention to the need for more effective management of pain, nausea, and other aversive side effects of state-of-the-art cancer therapy. One of the most promising approaches to effective management is nonpharmacologic intervention based on behavioral research and theory. The purpose of this review is to examine the effectiveness of behavioral intervention methods in the control of aversive side effects of cancer treatments. Fifty-four published studies using a variety of research designs were identified for review. Results indicated the following: 1) Behavioral intervention can effectively control anticipatory nausea and vomiting in adult and pediatric cancer patients undergoing chemotherapy; however, the evidence for the efficacy of behavioral intervention to control post chemotherapy nausea and vomiting is less clear. 2) Behavioral intervention integrating several behavioral methods can ameliorate anxiety and distress associated with invasive medical treatments. 3) Although a variety of behavioral methods have been shown to reduce acute treatment-related pain, there is increasing evidence that these methods are not equally effective. Hypnotic-like methods, involving relaxation, suggestion, and distracting imagery, hold the greatest promise for pain management. Unfortunately, research is scant on the use of behavioral intervention to control prolonged pain associated with invasive medical procedures. It is clear that the application of behavioral theory and methods has an important place in the care of patients undergoing invasive cancer treatments. PMID- 11390532 TI - Annual report to the nation on the status of cancer (1973 through 1998), featuring cancers with recent increasing trends. AB - BACKGROUND: The American Cancer Society, the National Cancer Institute (NCI), the North American Association of Central Cancer Registries, and the Centers for Disease Control and Prevention, including the National Center for Health Statistics (NCHS), collaborate to provide an annual update on cancer occurrence and trends in the United States. This year's report contains a special feature that focuses on cancers with recent increasing trends. METHODS: From 1992 through 1998, age-adjusted rates and annual percent changes are calculated for cancer incidence and underlying cause of death with the use of NCI incidence and NCHS mortality data. Joinpoint analysis, a model of joined line segments, is used to examine long-term trends for the four most common cancers and for those cancers with recent increasing trends in incidence or mortality. Statistically significant findings are based on a P value of.05 by use of a two-sided test. State-specific incidence and death rates for 1994 through 1998 are reported for major cancers. RESULTS: From 1992 through 1998, total cancer death rates declined in males and females, while cancer incidence rates declined only in males. Incidence rates in females increased slightly, largely because of breast cancer increases that occurred in some older age groups, possibly as a result of increased early detection. Female lung cancer mortality, a major cause of death in women, continued to increase but more slowly than in earlier years. In addition, the incidence or mortality rate increased in 10 other sites, accounting for about 13% of total cancer incidence and mortality in the United States. CONCLUSIONS: Overall cancer incidence and death rates continued to decline in the United States. Future progress will require sustained improvements in cancer prevention, screening, and treatment. PMID- 11390533 TI - Prevalence and risk factors for anal squamous intraepithelial lesions in women. AB - BACKGROUND: Anal cancers are thought to arise from squamous intraepithelial lesions in the anal canal, and women infected with human immunodeficiency virus-1 (HIV) may be at higher risk of anal cancer. Our aim was to determine the prevalence of human papillomavirus (HPV)-related abnormalities of the anal canal in women and to characterize risk factors for these lesions. METHODS: We evaluated HPV-related abnormalities in 251 HIV-positive and in 68 HIV-negative women. We completed physical examinations and obtained questionnaire data on medical history and relevant sexual practices. Univariate and adjusted relative risks (RRs) and 95% confidence intervals (CIs) were computed using the Mantel Haenszel procedure and regression techniques. All statistical tests were two sided. RESULTS: Abnormal anal cytology, including atypical squamous cells of undetermined significance, low-grade squamous intraepithelial lesions, or high grade squamous intraepithelial lesions (HSILs), was diagnosed in 26% of HIV positive and in 8% of HIV-negative women. HSILs were detected by histology or cytology in 6% of HIV-positive and in 2% of HIV-negative women. HIV-positive women showed increased risk of anal disease as the CD4 count decreased (P<.0001) and as the plasma HIV RNA viral load increased (P =.02). HIV-positive women with abnormal cervical cytology had an increased risk of abnormal anal cytology at the same visit (RR = 2.2; 95% CI = 1.4 to 3.3). Abnormal anal cytology in HIV positive women was associated with anal HPV RNA detected by the polymerase chain reaction and by a nonamplification-based test (RR = 4.3; 95% CI = 1.6 to 11). In a multivariate analysis, the history of anal intercourse and concurrent abnormal cervical cytology also were statistically significantly (P =.05) associated with abnormal anal cytology. CONCLUSIONS: HIV-positive women had a higher risk of abnormal anal cytology than did HIV-negative women with high-risk lifestyle factors. These data provide strong support for anoscopic and histologic assessment and careful follow-up of women with abnormal anal lesions. PMID- 11390534 TI - Age and adjuvant chemotherapy use after surgery for stage III colon cancer. AB - BACKGROUND: Randomized trials have established that 5-fluorouracil-based adjuvant chemotherapy following resection of stage III colon cancer reduces subsequent mortality by as much as 30%. However, the extent to which adjuvant therapy is used outside the clinical trial setting, particularly among the elderly, is unknown. METHODS: A retrospective cohort study utilizing the Surveillance, Epidemiology, and End Results/Medicare-linked database identified 6262 patients aged 65 years and older with resected stage III colon cancer. The primary outcome was chemotherapy use within 3 months of surgery, as ascertained from Medicare claims. We examined the extent to which age at diagnosis was associated with adjuvant chemotherapy usage, and we adjusted for potential confounding based on differences in other patient characteristics with the use of multiple logistic regression. All P values were two-sided. RESULTS: Age at diagnosis was the strongest determinant of chemotherapy: 78% of patients aged 65-69 years, 74% of those aged 70-74 years, 58% of those aged 75-79 years, 34% of those aged 80-84 years, and 11% of those aged 85-89 years received postoperative chemotherapy. The age trend remained pronounced after adjustment for potential confounding based on variation in patients' demographic and clinical characteristics and after exclusion of patients with any evident comorbidity (all P values <.001). CONCLUSIONS: Adjuvant chemotherapy for stage III colon cancer is used extensively, especially for patients under the age of 75 years. However, treatment rates decline dramatically with chronologic age. Because patients in their 70s and even 80s have a reasonable life expectancy, further efforts are needed to ensure that elderly patients have the opportunity to make informed decisions regarding this potentially curative treatment. PMID- 11390535 TI - Detecting colorectal cancer in stool with the use of multiple genetic targets. AB - BACKGROUND: Colorectal cancer cells are shed into the stool, providing a potential means for the early detection of the disease using noninvasive approaches. Our goal was to develop reliable, specific molecular genetic tests for the detection of colorectal cancer in stool samples. METHODS: Stool DNA was isolated from paired stools and primary tumor samples from 51 colorectal cancer patients. Three genetic targets-TP53, BAT26, and K-RAS-were used to detect tumor associated mutations in the stool prior to or without regard to the molecular analyses of the paired tumors. TP53 gene mutations were detected with a mismatch ligation assay that detects nine common p53 gene mutations. Deletions within the BAT26 locus were detected by a modified solid-phase minisequencing method. Mutations in codons 12 and 13 of K-RAS were detected with a digital polymerase chain reaction-based method. RESULTS: TP53 gene mutations were detected in the tumor DNA of 30 patients, all of whom had the identical TP53 mutation in their stools. Tumors from three patients contained a noninherited deletion at the BAT26 locus, and the same alterations were identified in these patients' stool specimens. Nineteen of 50 tumors tested had a K-RAS mutation; identical mutations were detected in the paired stool DNA samples from eight patients. In no case was a mutation found in stool that was not also present in the primary tumor. Thus, the three genetic markers together detected 36 (71%) of 51 patients (95% confidence interval [CI] = 56% to 83%) with colorectal cancer and 36 (92%) of 39 patients (95% CI = 79% to 98%) whose tumors had an alteration. CONCLUSION: We were able to detect the majority of colorectal cancers by analyzing stool DNA for just three genetic markers. Additional work is needed to determine the specificity of these genetic tests for detecting colorectal neoplasia in asymptomatic patients and to more precisely estimate the prevalence of the mutations and sensitivity of the assay. PMID- 11390536 TI - Telomerase suppression by chromosome 6 in a human papillomavirus type 16 immortalized keratinocyte cell line and in a cervical cancer cell line. AB - BACKGROUND: High-risk human papillomavirus (HPV) types play a major role in the development of cervical cancer in vivo and can induce immortalization of primary human keratinocytes in vitro. Activation of the telomere-lengthening enzyme telomerase constitutes a key event in both processes. Because losses of alleles from chromosome 6 and increased telomerase activity have been observed in high grade premalignant cervical lesions, we analyzed whether human chromosome 6 harbors a putative telomerase repressor locus that may be involved in HPV mediated immortalization. METHODS: Microcell-mediated chromosome transfer was used to introduce chromosomes 6 and 11 to the in vitro generated HPV type 16 (HPV16)-immortalized keratinocyte cell line FK16A and to the in vivo derived HPV16-containing cervical cancer cell line SIHA: Hybrid clones were analyzed for growth characteristics, telomerase activity, human telomerase reverse transcriptase (hTERT) and HPV16 E6 expression, and telomere length. FK16A hybrid clones were also transduced with an hTERT-containing retrovirus to examine the effect of ectopic hTERT expression on growth. Statistical tests were two-sided. RESULTS: Introduction of human chromosome 6 but not of chromosome 11 to both cell lines yielded hybrid cells that demonstrated crisis-like features (i.e., enlarged and flattened morphology, vacuolation, and multinucleation) and underwent growth arrest after a marked lag period. In the chromosome 6 hybrid clones analyzed, telomerase activity and hTERT messenger RNA (mRNA) expression were statistically significantly reduced compared with those in the chromosome 11 hybrid clones (for telomerase activity, P =.004 for the FK16A hybrids and P =.039 for the SiHa hybrids; for hTERT mRNA expression, P =.003 for the FK16A hybrids). The observed growth arrest was associated with telomeric shortening. Ectopic expression of hTERT in FK16A cells could prevent the telomeric shortening-based growth arrest induced by chromosome 6. CONCLUSIONS: Chromosome 6 may harbor a repressor of hTERT transcription, the loss of which may be involved in HPV-mediated immortalization. PMID- 11390537 TI - Experimental confirmation of a distinctive diffraction pattern in hair from women with breast cancer. PMID- 11390538 TI - Prostate cancer: different incidence but not mortality trends within two areas of Tuscany, Italy. PMID- 11390539 TI - Re: re: role of the insulin-like growth factors in cancer development and progression. PMID- 11390540 TI - Re: cellular telephones and cancer--a nationwide cohort study in Denmark. PMID- 11390542 TI - Re: cellular telephones and cancer--a nationwide cohort study in Denmark. PMID- 11390544 TI - Prediction of myocardial infarction versus cardiac death by gated myocardial perfusion SPECT: risk stratification by the amount of stress-induced ischemia and the poststress ejection fraction. AB - The combination of myocardial perfusion and poststress ejection fraction (EF) provides incremental prognostic information. This study assessed predictors of nonfatal myocardial infarction (MI) versus cardiac death (CD) by gated myocardial SPECT and examined the value of integrating the amount of ischemia and poststress EF data in risk stratification. METHODS: We identified 2,686 patients who underwent resting (201)Tl/stress (99m)Tc-sestamibi gated SPECT and were monitored for >1 y. Patients who underwent revascularization < or = 60 d after the nuclear test were censored from the prognostic analysis. Visual scoring of perfusion images used 20 segments and a scale of 0--4. Poststress EF was automatically generated. RESULTS: Cox regression analysis showed that after adjusting for prescan data, the most powerful predictor of CD was poststress EF, whereas the best predictor of MI was the amount of ischemia (summed difference score [SDS]). Integration of the EF and SDS yielded effective stratification of patients into low-, intermediate-, and high-risk subgroups. Patients with EF >50% and a large amount of ischemia were at intermediate risk (2%--3%), whereas those with mild or moderate ischemia were at low risk of CD (<1%/y). Patients with EF between 30% and 50% were at intermediate risk even in the presence of only mild or moderate ischemia. In patients with EF <30%, the CD rate was high (>4%/y) irrespective of the amount of ischemia. CONCLUSION: Poststress EF is the best predictor of CD, whereas the amount of ischemia is the best predictor of nonfatal MI. Integration of perfusion and function data improves stratification of patients into low, intermediate, and high risk of CD. PMID- 11390545 TI - Low-dose dobutamine electrocardiograph-gated myocardial SPECT for identifying viable myocardium: comparison with dobutamine stress echocardiography and PET. AB - The identification of severely dysfunctional but viable myocardium is of particular importance for the selection of patients with depressed left ventricular function who will benefit from coronary revascularization. Assessment of inotropic reserve with dobutamine has recently been used for this purpose. This study compared the accuracy of low-dose dobutamine stress gated myocardial SPECT (DS SPECT) with the accuracy of dobutamine stress echocardiography (DSE) and resting perfusion SPECT for the identification of viable myocardium in patients with previous myocardial infarction. METHODS: Resting and low-dose dobutamine (7.5 microg/kg/min) gated (99m)Tc-tetrofosmin SPECT and echocardiography and resting (18)F-FDG PET were prospectively studied in 23 patients with previous myocardial infarction and severely depressed regional function. Twenty-one of them were successfully studied with each technique. The left ventricular wall was divided into 14 segments to assess wall motion using a 5-point scale. PET viability was defined as FDG uptake >/= 50% of the maximum uptake in a region with normal wall motion. For DS SPECT and DSE studies, viable myocardium was defined as hypokinetic areas with > or = 1 point improvement in wall motion. For resting perfusion SPECT, viable myocardium was defined as hypokinetic areas with a relative uptake > or = 50% of the maximum uptake. RESULTS: Of a total of 294 segments, 55 had severe resting dyskinesis. Thirty four segments were identified as viable on FDG PET, and 21 segments were identified as nonviable. Eleven segments were inadequately visualized with DSE, including 5 segments in the apex. Sensitivities (78% vs. 76%) and specificities (94% vs. 100%) were similar for DSE and DS SPECT, with a concordance of 86% (kappa = 0.72). DS SPECT and perfusion SPECT did not significantly differ with respect to sensitivities (76% vs. 85%, respectively). However, specificity was significantly higher for DS SPECT than for perfusion SPECT (100% vs. 52%, respectively, P < 0.05). CONCLUSION: This study indicated that DS SPECT correlates well with DSE in the assessment of viability. In addition, gated SPECT can evaluate regional wall motion, even in areas inadequately assessed by echocardiography. DS SPECT may also provide additional information for identifying viable myocardium, which is often overestimated by routine perfusion scans. PMID- 11390546 TI - Myocardial tracking, a new method to calculate ejection fraction with gated SPECT: validation with (201)Tl versus planar angiography. AB - Left ventricular ejection fraction (LVEF) and viability are essential variables for the prognosis of myocardial infarction and can be measured simultaneously by (201)Tl gated SPECT; however, most algorithms tend to underestimate LVEF. This study aimed to evaluate a new myocardial tracking algorithm, MyoTrack (MTK), for automatic LVEF calculation. METHODS: A rest/redistribution (20 min/4 h) (201)Tl gated SPECT protocol followed immediately by a (99m)Tc equilibrium radionuclide angiography (ERNA) was performed in 75 patients with history of myocardial infarction. Quality of myocardial uptake was evaluated from count statistics and automatic quantification of defect sizes and severities (CardioMatch). LVEFs were calculated both with Germano's quantitative gated SPECT (QGS) algorithm and with MTK. Briefly, the originality of this algorithm resides in the unique end diastole segmentation, matching to a template and motion field tracking throughout the cardiac cycle. RESULTS: ERNA LVEF averaged 33% +/- 14%. QGS significantly underestimated this value at 20 min (30% +/- 13%, P < 0.001) and at 4 h (30% +/- 13%, P < 0.0001). By contrast, MTK did not miscalculate LVEF at 20 min (34% +/- 14%, probability value was not significant) though a similar underestimation occurred at 4 h (31% +/- 13%, P < 0.02). Individual differences between early and late gated SPECT values and differences between gated SPECT and ERNA values did not correlate with the extension of perfusion defects, count statistics, or heart rate. CONCLUSION: MTK algorithm accurately calculates LVEF on early/high-count images compared with ERNA [corrected], even in patients with severe perfusion defects, but tends to underestimate LVEF on delayed/low-contrast images, as other algorithms do. PMID- 11390547 TI - Ictal hyperperfusion of cerebellum and basal ganglia in temporal lobe epilepsy: SPECT subtraction with MRI coregistration. AB - The ictal hyperperfusion (compared with the interictal state) of the cerebellum and basal ganglia has not been investigated systematically in patients with temporal lobe epilepsy (TLE). Their ictal perfusion patterns were analyzed in relation to temporal and frontal hyperperfusion during TLE seizures using SPECT subtraction. METHODS: Thirty-three TLE patients had interictal and ictal SPECT, video-electroencephalographic (EEG) monitoring, and volumetric MRI. SPECT subtraction with MRI coregistration was performed using commercial software. The presence of ictal hyperperfusion was determined in the ipsilateral and contralateral temporal lobe, frontal lobe, cerebellum, and basal ganglia. RESULTS: All patients showed ictal hyperperfusion in the temporal lobe of seizure origin. Vermian cerebellar hyperperfusion (CH) was observed in 26 patients (78.8%) and hemispheric CH was found in 25 (75.8%). Compared with the side of the epileptogenic temporal lobe, there were 7 patients with ipsilateral hemispheric CH (28.0%), 15 with contralateral hemispheric CH (60.0%), and 3 with bilateral hemispheric CH (12.0%). CH was observed more frequently in patients with additional frontal hyperperfusion (14/15, 93.3%; 2 ipsilateral to the seizure focus, 10 contralateral, and 2 bilateral) than in patients without frontal hyperperfusion (11/18, 61.1%). Among 18 patients with temporal hyperperfusion without frontal hyperperfusion, 11 patients showed hemispheric CH (5 ipsilateral to seizure focus, 5 contralateral, 1 bilateral). Hyperperfusion in the basal ganglia (BGH) was seen in 11 of the 15 patients with temporal and frontal hyperperfusion (73.3%) and in 11 of the 18 with only temporal hyperperfusion (61.1%). In 17 patients with unilateral BGH (13 ipsilateral to the seizure focus, 4 contralateral), CH contralateral to the BGH was observed in 14 (82.5%), CH ipsilateral to the BGH was found in 2 (11.8%), and CH bilateral to the BGH was found in 1 (5.9%). CONCLUSION: During TLE seizures, hemispheric CH occurred not only in contralateral but also in ipsilateral or bilateral cerebellar hemispheres to the side of seizure origin. Although temporal lobe origin seizures associated with additional frontal hyperperfusion produced more frequent hemispheric CH, seizures showing only temporal hyperperfusion without frontal hyperperfusion could produce BGH and CH. To determine the side of hemispheric CH, the most important factor appears to be the side of BGH, not the side of seizure origin. PMID- 11390548 TI - Embolic distribution through patent foramen ovale demonstrated by (99m)Tc-MAA brain SPECT after Valsalva radionuclide venography. AB - Cryptogenic stroke might relate to paradoxical embolism stemming from right-to left shunt caused by patent foramen ovale (PFO). We performed radionuclide venography using the Valsalva maneuver, followed by (99m)Tc-macroaggregated albumin (MAA) brain SPECT to investigate the fate of emboli originating from the lower extremities. METHODS: Ten patients (9 men, 1 woman; mean age, 61 +/- 17 y) with PFO underwent radionuclide venography with and without the Valsalva maneuver on the whole-body image, followed by brain SPECT with (99m)Tc-MAA to determine the cortical uptake that would detect right-to-left shunt. After counts in each region of interest (ROI) were normalized by comparison with the averaged count, the distribution of MAA was compared with that of (99m)Tc-hexamethyl propyleneamine oxime (HMPAO) brain SPECT by drawing ROIs on frontal, temporoparietal (anterior circulation territory), occipital, and cerebellar areas (posterior circulation territory). RESULTS: The thyroid on the whole-body scan was visualized after the Valsalva maneuver in 2 of the 10 patients. In 7 of 10 patients, 56 ROIs in the visualized cortical uptake showed that the distribution of MAA correlated well with that of HMPAO according to the equation: HMPAO = 71.21 + 1.71 x MAA, (r = 0.575, P < 0.01). The excess difference in the relative counts in the posterior over anterior circulation territory was 5.6% and 16.1% of the HMPAO and MAA values, respectively. CONCLUSION: Brain SPECT with (99m)Tc-MAA was more sensitive than thyroid visualization in detecting right-to-left shunt. The excess flow in the posterior cerebral circulation indicated an increased likelihood of cerebral emboli originating from the lower extremities and indicated that the flow difference between HMPAO and MAA probably resulted from poor linearization of HMPAO in the high-flow area. PMID- 11390549 TI - (133)Xe SPECT cerebral blood flow study in a healthy population: determination of T-scores. AB - Dementia is becoming a major health problem as the population of the Northern Hemisphere ages. Early differential diagnosis between normal cognitive decline and dementia is particularly difficult. If psychometric evaluation can contribute to the diagnosis, quantitative cerebral functional imaging would play an important role. We therefore proposed, first, to constitute a normative dataset that could later be used to identify subjects at risk for neurodegenerative processes and, second, to describe the risk of abnormal global cerebral blood flow (gCBF) by defining categories based on the standardized cutoff scores of a young, healthy population (T-score). METHODS: Of a total of 203 healthy volunteers, 187 were included in the protocol, which included evaluation of medical history, neurologic and neuropsychologic status, and body composition; analysis of blood; and measurement of gCBF by means of (133)Xe SPECT. RESULTS: With ANOVA analysis using age and sex as between-subject factors and gCBF as a within-subject factor, a significantly higher gCBF was found in women than in men. In addition, a linear reduction as a function of age was observed for both sexes (-0.3%/y). T-score was determined for the 18- to 28-y-old age group, for whom gCBF was found to be 46.7 +/- 5.1 mL/min/100 g tissue in men and 49.0 +/- 5.0 mL/min/100 g tissue in women. The age-dependent decrease could thus be expressed in T-scores and, in the 29- to 38-y-old, 39- to 48-y-old, and >48-y-old age groups, averaged -0.63, -1.29, and -1.92, respectively, in men and -0.63, 0.83, and-2.40, respectively, in women. Cognitive performance, body composition, and blood analysis revealed the expected significant effects from sex and age. CONCLUSION: The large-scale reference database of gCBF measurements constituted from a healthy, well-controlled population enabled age and sex stratification, which showed significant differences between the sexes and a significant decline as a function of age. T-scores were determined and warrant further studies on the prospective identification of early dementia by (133)Xe SPECT in elderly individuals. PMID- 11390550 TI - Incremental prognostic value of RNA ejection fraction measurements during pharmacologic stress testing: a comparison with clinical and perfusion variables. AB - This investigation examined the prognostic power of first-pass radionuclide angiocardiography (RNA) ejection fraction compared with clinical information and myocardial perfusion imaging in patients undergoing pharmacologic stress testing. The value of RNA and myocardial perfusion imaging in predicting death or nonfatal myocardial infarction (MI) is well established. However, limited information exists on the usefulness of combined myocardial perfusion imaging and RNA to predict prognosis, especially in patients undergoing pharmacologic stress testing. METHODS: We identified 240 patients who underwent pharmacologic stress testing with myocardial perfusion imaging and combined RNA. The patients were followed for a mean of 1.4 y. Cox proportional hazards models were used to assess the value in predicting death and MI. Multivariable models were generated to assess the independent incremental predictive value of clinical and nuclear imaging variables. Kaplan-Meier survival and event-free survival estimates were examined in patients with low (< or = 45%) versus high (>45%) ejection fractions. RESULTS: Clinical information, myocardial perfusion imaging, and RNA ejection fraction were significant predictors of the death/MI composite outcome (chi(2) = 7.4, 14.0, and 21.8, respectively). The addition of myocardial perfusion imaging to the clinical information provided incremental prognostic information (chi(2) = 15.2). The addition of RNA ejection fraction provided further predictive information (chi(2) = 22.5). However, when RNA ejection fraction was first added to the clinical information, myocardial perfusion imaging had no incremental prognostic value. CONCLUSION: For hard cardiac events, RNA ejection fraction provides prognostic information besides that provided by clinical and myocardial perfusion imaging. In patients who cannot exercise and are undergoing noninvasive evaluation with pharmacologic stress testing and myocardial perfusion imaging, ejection fraction should be measured simultaneously for risk assessment optimization. PMID- 11390551 TI - (99m)Tc-MDP scintigraphic findings in children with leukemia: value of early and delayed whole-body imaging. AB - The purpose of this study was to reveal the bone scan abnormalities in children with leukemia and to show the value of whole-body scanning in early and delayed phases. METHODS: From a database of all patients with a diagnosis of leukemia from January 1990 to April 2000, 12 children (9 male, 3 female; mean age, 8.0 y; age range, 4.7--13.2 y) were identified for whom the diagnosis of leukemia was suggested on the basis of bone scans obtained as part of the initial work-up for unexplained skeletal pain. Early and delayed whole-body bone scans and radiographs were reviewed retrospectively. Areas of abnormal uptake on early and delayed phases were categorized into locations: metaphysis--diaphysis--epiphysis (MDE), pelvis, ribs, spine, and others. MDE lesions included abnormalities in the metaphysis extending into the diaphysis for some length: metaphysis/diaphysis, metaphysis only, diaphysis only, epiphysis only, and the entire bone. Pelvic and spine lesions were further characterized as focal or diffuse. RESULTS: Ten patients had lesions in 2 or more locations on both phases. Two patients had multiple lesions on the early scans but only rib lesions on the delayed scans. Lesions correlated with symptomatic sites in 8 patients on the delayed scans and in 11 patients on the early scans. The most common sites of abnormalities on the delayed scans were metaphyseal/diaphyseal, pelvis (focal), and ribs. The most common locations of lesions on the early scans were metaphyseal/diaphyseal, pelvis (diffuse or focal), and spine. More metaphyseal/diaphyseal lesions were seen on the early scans than on the delayed scans. Diffuse involvement of the pelvis and spine was seen only on the early phase. However, rib lesions were seen more frequently on the delayed scan. CONCLUSION: Early whole-body imaging in conjunction with delayed whole-body scanning may enhance the diagnostic accuracy of bone scanning in the evaluation of children with skeletal pain of obscure etiology, such as that associated with leukemia. PMID- 11390552 TI - The diagnostic role of radionuclide imaging in evaluation of patients with nonhypersecreting adrenal masses. AB - The aim of this study was to evaluate the role of radionuclide imaging in the characterization of nonhypersecreting adrenal masses. METHODS: A total of 54 patients (19 men, 35 women; mean age, 50 +/- 16 y) with nonhypersecreting unilateral adrenal tumors that had been originally detected on CT or MRI underwent adrenal scintigraphy using different radiotracers. None of the patients showed specific symptoms of adrenal hypersecretion. Screening tests for excess cortical and medullary products showed normal adrenal hormone levels. Radionuclide studies (n = 73) included (131)I-norcholesterol (n = 24), (131)I metaiodobenzylguanidine (MIBG) (n = 23), and (18)F-FDG PET (n = 26) scans. RESULTS: Histology after surgery (n = 31) or adrenal biopsy (n = 23) was obtained. Adrenal lesions were represented by 19 adenomas, 4 cysts, 1 myelolipoma, 1 neurinoma, 2 ganglioneuromas, 5 pheochromocytomas, 4 pseudotumors, 6 carcinomas, 2 sarcomas, and 10 metastases (size range, 1.5- to 5-cm diameter; mean, 4.9 +/- 3.1 cm). For norcholesterol imaging, diagnostic sensitivity, specificity, and accuracy were 100%, 71%, and 92%, respectively; the positive predictive value (PPV) of the norcholesterol scan to characterize an adrenal mass as an adenoma was 89%, whereas the corresponding negative predictive value (NPV) to rule out this type of tumor was 100%. For MIBG imaging, diagnostic sensitivity, specificity, and accuracy were 100%, 94%, and 96%, respectively; the PPV of the MIBG scan to characterize an adrenal mass as a medullary chromaffin tissue tumor was 83%, whereas the corresponding NPV to rule out this type of tumor was 100%. For FDG PET, diagnostic sensitivity, specificity, and accuracy were 100%, 100%, and 100%, respectively; the PPV of FDG PET to characterize an adrenal mass as a malignant tumor was 100%, whereas the corresponding NPV to rule it out was 100%. Furthermore, in 7 patients with malignant adrenal tumors, FDG whole-body scanning revealed extra-adrenal tumor sites (n = 29), allowing an accurate diagnosis of the disease's stage using a single-imaging technique. CONCLUSION: In patients with nonhypersecreting adrenal masses, radionuclide adrenal imaging, using specific radiopharmaceuticals such as norcholesterol, MIBG, and FDG, may provide significant functional information for tissue characterization. Norcholesterol and MIBG scans are able to detect benign tumors such as adenoma and pheochromocytoma, respectively. Conversely, FDG PET allows for recognition of malignant adrenal lesions. Therefore, adrenal scintigraphy is recommended for tumor diagnosis and, hence, for appropriate treatment planning, particularly when CT or MRI findings are inconclusive for lesion characterization. PMID- 11390553 TI - Role of molecular imaging in management of nonhypersecreting adrenal masses. PMID- 11390554 TI - Therapy of metastatic bone pain. AB - Bone metastasis is a common sequella of solid malignant tumors such as prostate, breast, lung, and renal cancers, which can lead to various complications, including fractures, hypercalcemia, and bone pain, as well as reduced performance status and quality of life. A multidisciplinary approach is usually required not only to address the etiology of the pain and its complicating factors but also to treat the patient appropriately. Currently, the treatment of bone pain remains palliative at best with systemic therapy (analgesics, hormones, chemotherapy, steroids, and bisphosphonates) as well as local treatments (such as surgery, nerve blocks, and external beam radiation). However, many of these treatments are limited in their efficacy or duration and have significant side effects that seriously limit the cancer patient's quality of life. Various radiopharmaceuticals have shown good efficacy in relieving bone pain secondary to bone metastasis. This systemic form of metabolic radiotherapy is simple to administer and complements other treatment options. This has been associated with improved mobility in many patients, reduced dependence on narcotic and non narcotic analgesics, improved performance status and quality of life, and, in some studies, improved survival. Additional radiopharmaceuticals are under investigation and appear promising. All of these agents, although comprising different physical and chemical characteristics, offer certain advantages in that they are simple to administer, are well tolerated by the patient if used appropriately, and can be used alone or in combination with the other forms of treatment. PMID- 11390555 TI - Outpatient treatment with (131)I-anti-B1 antibody: radiation exposure to family members. AB - The Nuclear Regulatory Commission (NRC) regulations that govern release of patients administered radioactive material have been revised to include dose based criteria in addition to the conventional activity-based criteria. A licensee may now release a patient if the total effective dose equivalent to another individual from exposure to the released patient is not likely to exceed 5 mSv (500 mrem). The result of this dose-based release limit is that now many patients given therapeutic amounts of radioactive material no longer require hospitalization. This article presents measured dose data for 26 family members exposed to 22 patients treated for non-Hodgkin's lymphoma with (131)I-anti-B1 antibody after their release according to the new NRC dose-based regulations. METHODS: The patients received administered activities ranging from 0.94 to 4.77 GBq (25--129 mCi). Family members were provided with radiation monitoring devices (film badges, thermoluminescent or optically stimulated luminescent dosimeters, or electronic digital dosimeters). Radiation safety personnel instructed the family members on the proper wearing and use of the devices. Instruction was also provided on actions recommended to maintain doses to potentially exposed individuals as low as is reasonably achievable. RESULTS: Family members wore the dosimeters for 2--17 d, with the range of measured dose values extending from 0.17 to 4.09 mSv (17--409 mrem). The average dose for infinite time based on dosimeter readings was 32% of the predicted doses projected to be received by the family members using the NRC method provided in regulatory guide 8.39. CONCLUSION: Therapy with (131)I-anti-B1 antibody can be conducted on an outpatient basis using the established recommended protocol. The patients can be released immediately with confidence that doses to other individuals will be below the 5-mSv (500 mrem) limit. PMID- 11390556 TI - Investigation of patient release criteria. PMID- 11390557 TI - Rapid imaging of experimental colitis with (99m)Tc-interleukin-8 in rabbits. AB - Radiolabeled autologous leukocytes (WBCs) are the gold standard for imaging inflammatory bowel disease (IBD). For the rapid and adequate management of patients with IBD, there is need for a new agent at least as good as radiolabeled WBCs, but easier to prepare and without its inherent risks. In this study, the potential of interleukin-8 (IL-8) labeled with (99m)Tc using hydrazinonicotinamide (HYNIC) to image IBD was investigated in a rabbit model of acute colitis and compared with that of (99m)Tc-HMPAO-labeled granulocytes. METHODS: In rabbits with chemically induced acute colitis, inflammatory lesions were scintigraphically visualized after injection of either IL-8 or purified granulocytes, both labeled with (99m)Tc. Gamma camera images were acquired at 2 min and at 1, 2, and 4 h after injection. Four hours after injection, the rabbits were killed, and the uptake of the radiolabel in the dissected tissues was determined. The dissected colon was imaged and the inflammatory lesions were scored macroscopically. For each affected colon segment, the colitis index (affected colon-to-normal colon uptake ratio, CI) was calculated and correlated with the macroscopically scored severity of inflammation. RESULTS: Both agents visualized the colitis within 1 h after injection. (99m)Tc-HYNIC-IL-8 images of the colonic abnormalities were more accurate and the intensity of uptake in the affected colon continuously increased until 4 h after injection, whereas no further increase 1 h after injection was noticed scintigraphically for (99m)Tc HMPAO-granulocytes. The absolute uptake in the affected colon was much higher for IL-8 than for the radiolabeled granulocytes with the percentage injected dose per gram (%ID/g) 0.41 +/- 0.04 %ID/g and 0.09 +/- 0.05 4 %ID/g h after injection, respectively. With increasing severity, the CI at 4 h after injection for (99m)Tc HYNIC-IL-8 was 4.4 +/- 0.6, 13.5 +/- 0.5, and 25.8 +/- 1.0; for granulocytes, the CI at 4 h after injection was 1.5 +/- 0.1, 3.4 +/- 0.2, and 6.4 +/- 0.5, respectively. The CI correlated with the severity of the inflammation (r = 0.95, P < 0.0001 for IL-8; r = 0.95, P < 0.0001 for granulocytes). CONCLUSION: Within 1 h after injection, visualization of the extent of colonic inflammation in vivo was possible with (99m)Tc-HYNIC-IL-8 and (99m)Tc-HMPAO-granulocytes. Within 2 h after injection, (99m)Tc-IL-8 allowed a good evaluation, and within 4 h after injection, a meticulous evaluation of the severity of IBD. Although (99m)Tc-HMPAO granulocytes were able to delineate the extent of IBD within 2 h after injection, an accurate estimation of severity of inflammation was not possible. (99m)Tc HYNIC-IL-8 is an inflammation-imaging agent that showed promising results in this study. (99m)Tc-IL-8 can be prepared off-the-shelf and yields excellent imaging with high target-to-background ratios. PMID- 11390558 TI - A fast nonlinear method for parametric imaging of myocardial perfusion by dynamic (13)N-ammonia PET. AB - A parametric image of myocardial perfusion (mL/min/g) is a quantitative image generated by fitting a tracer kinetic model to dynamic (13)N-ammonia PET data on a pixel-by-pixel basis. There are several methods for such parameter estimation problems, including weighted nonlinear regression (WNLR) and a fast linearizing method known as Patlak analysis. Previous work showed that sigmoidal networks can be used for parameter estimation of mono- and biexponential models. The method used in this study is a hybrid of WNLR and sigmoidal networks called nonlinear regression estimation (NRE). The purpose of the study is to compare NRE with WNLR and Patlak analysis for parametric imaging of perfusion in the canine heart by (13)N-ammonia PET. METHODS: A simulation study measured the statistical performance of NRE, WNLR, and Patlak analysis for a probabilistic model of time activity curves. Four canine subjects were injected with 740 MBq (13)N-ammonia and scanned dynamically. Images were reconstructed with filtered backprojection and resliced into short-axis cuts. Parametric images of a single midventricular plane per subject were generated by NRE, WNLR, and Patlak analysis. Small regions of interest (ROIs) were drawn on each parametric image (8 ROIs per subject for a total of 32). RESULTS: For the simulation study, the median absolute value of the relative error for a perfusion value of 1.0 mL/min/g was 16.6% for NRE, 17.9% for WNLR, 19.5% for Patlak analysis, and 14.5% for an optimal WNLR method (computable by simulation only). All methods are unbiased conditioned on a wide range of perfusion values. For the canine studies, the least squares line fits comparing NRE (y) and Patlak analysis (z) with WNLR (x) for all 32 ROIs were y = 1.02x - 0.028 and z = 0.90x + 0.019, respectively. Both NRE and Patlak analysis generate 128 x 128 parametric images in seconds. CONCLUSION: The statistical performance of NRE is competitive with WNLR and superior to Patlak analysis for parametric imaging of myocardial perfusion. NRE is a fast nonlinear alternative to Patlak analysis and other fast linearizing methods for parametric imaging. NRE should be applicable to many other tracers and tracer kinetic models. PMID- 11390559 TI - Myocardial kinetics of the (11)C-labeled enantiomers of the Ca(2+) channel inhibitor S11568: an in vivo study. AB - Ca(2+) channels play a key role in the basic working of the heart. There is one particular type of Ca(2+) channel in cardiac cells (L-type) whose gating is affected in different ways by beta-adrenoceptors and 1,4-dihydropyridines. In this study, we used ex vivo studies and PET to evaluate and compare the myocardial kinetics of the enantiomers labeled with (11)C (the more active: S12968, absolute configuration S; the less active: S12967, absolute configuration R) of the L-type Ca(2+) channel antagonist S11568 (3-ethyl 5-methyl (+/-)-2-[(2 (2-aminoethoxy)ethoxy) methyl]-4-(2,3-dichlorophenyl)-6-methyl-1,4 dihydropyridine-3,5-dicarboxylate). METHODS: [(11)C]S12968 was injected into the tail vein of rats (0.22 kBq--5.92 MBq) to assess the relationship between injected dose and myocardial uptake. A series of 5 rats was pretreated with 4 micromol unlabeled S12968 5 min before injection of 2.2 kBq [(11)C]S12968. In another series of 5 rats, unlabeled S12698 (4 micromol) was injected 5 min after injection of 2.2 kBq [(11)C]S12968. The animals were killed 15 min later, and the myocardial radioactivity was assessed in a gamma well counter. Beagle dogs received injections of 5-15 nmol [(11)C]S12968 or [(11)C]S12967 and were imaged with PET. Presaturation and displacement experiments using 2 micromol/kg unlabeled S12968 or 6 mol/kg S12967 were performed. RESULTS: In rats, a statistically significant relationship between myocardial uptake and injected dose of S12968 was observed. Pretreatment or displacement with unlabeled S12968 reduced myocardial radioactivity by 75% and 70%, respectively. In dogs, after injection of 5 nmol of each enantiomer, myocardial radioactivity plateaued within 3 min and the clearance from blood was rapid. Injection of 13--15 nmol [(11)C]S12968 led to a higher myocardial uptake and a more rapid washout, which were related to an increased coronary blood flow as shown by the linear relationship between k(1)--an estimate of coronary blood flow--and the mass of S12968 injected. Presaturation and displacement experiments showed that 70%--80% of S12968 binding was specific. This specificity was not observed with S12967. Plasma metabolite analysis showed that 70% of the compound was unchanged 20 min after injection. CONCLUSION: These results show the feasibility of imaging myocardial L-type Ca(2+) channels in vivo using [(11)C]S12968. PMID- 11390560 TI - Blind separation of cardiac components and extraction of input function from H(2)(15)O dynamic myocardial PET using independent component analysis. AB - The independent component analysis (ICA) method is suggested to be useful for separation of the ventricles and the myocardium and for extraction of the left ventricular input function from the dynamic H(2)(15)O myocardial PET. The ICA generated input function was validated with the sampling method, and the myocardial blood flow (MBF) calculated with this input function was compared with the microsphere results. METHODS: We assumed that the elementary activities of the ventricular pools and the myocardium were spatially independent and that the mixture of them composed dynamic PET image frames. The independent components were estimated by recursively minimizing the mutual information (measure of dependence) between the components. The ICA-generated input functions were compared with invasively derived arterial blood samples. Moreover, the regional MBF calculated using the ICA-generated input functions and single-compartment model was correlated with the results obtained from the radiolabeled microspheres. RESULTS: The ventricles and the myocardium were successfully separated in all cases within a short computation time (<15 s). The ICA-generated input functions displayed shapes similar to those obtained by arterial sampling except that they had a smoother tail than those obtained by sampling, which meant that ICA removed the statistical noise from the time--activity curves. The ICA generated input function showed a longer time delay of peaks than those obtained by arterial sampling. MBFs estimated using the ICA-generated input functions ranged from 1.10 to approximately 2.52 mL/min/g at rest and from 1.69 to approximately 8.00 mL/min/g after stress and correlated well with those calculated with microspheres (y = 0.45 + 0.98x; r = 0.95, P < 0.000). CONCLUSION: ICA, a rapid and reliable method for extraction of the pure physiologic components, was a valid and useful method for quantification of the regional MBF using H(2)(15)O PET. PMID- 11390561 TI - Feasibility of dual-isotope coincidence/single-photon imaging of the myocardium. AB - Hybrid PET scanners offer the possibility of obtaining myocardial viability information from coincidence imaging of the positron emitter (18)F-FDG and perfusion measurements from a single-photon tracer-potentially simultaneously. This new approach is less costly and more readily available than dedicated PET and offers potential for improved FDG resolution and sensitivity compared with SPECT with 511-keV collimators. Simultaneous imaging of the coincidence and single-photon events offers the further advantages of automatic image registration and reduced imaging time. However, the feasibility of simultaneous coincidence/single-photon imaging or even immediately sequential imaging is unknown. In this study, the potential of using standard low-energy high resolution (LEHR) collimators with hybrid PET to obtain coincidence and SPECT data was assessed. METHODS: Phantom and human studies were performed to investigate the effect of LEHR collimators on FDG coincidence imaging with a hybrid PET system, the effect of the presence of (99m)Tc during FDG coincidence imaging with LEHR collimators, and the effect of the presence of FDG during (99m)Tc SPECT imaging. RESULTS: FDG images were somewhat degraded (a measure of myocardial nonuniformity increased 10%) with LEHR collimators. With 148 MBq (4 mCi) (99m)Tc present during FDG imaging of a phantom, image quality was maintained and the number of detected coincidences changed by <5%. With (99m)Tc/(18)F whole-body ratios of 7:1, crosstalk from (18)F photons accounted for the majority of counts in the (99m)Tc SPECT images and resulted in severe artifacts. The artifacts were decreased with a simple crosstalk correction scheme but remained problematic. CONCLUSION: (99m)Tc/(18)F ratios of at least 9:1 and state-of-the-art reconstruction and crosstalk correction are likely to be required to perform immediately sequential coincidence/single-photon imaging of the myocardium with clinically useful results. Additional challenges remain before simultaneous imaging of coincidence events and single photons can be realized in practice. PMID- 11390562 TI - A synthetic macromolecule for sentinel node detection: (99m)Tc-DTPA-mannosyl dextran. AB - We report the synthesis and preliminary biologic testing of a synthetic macromolecule, (99m)Tc-diethylenetriaminepentaacetic acid (DTPA)--mannosyl dextran, for sentinel node detection. METHODS: Synthesis started with a 2-step process that attaches a high density of amino-terminated leashes to a dextran backbone. Allyl-bromide was reacted with pharmaceutical-grade dextran to yield allyl-dextran. After diafiltration with water, filtration, and lyophilization, the product was reacted with aminoethanethiol and ammonium persulfate. The resulting amino-conjugated dextran was dialyzed, filtered, and lyophilized. The mixed anhydride method was used to attach DTPA; after dialysis, filtration, and lyophilization, 2-imino-2-methoxyethyl-1-D-mannose was used to attach the receptor substrate. The molecular diameter was measured by dynamic light scattering. Amino, mannose, and DTPA densities were measured by trinitrobenzene sulfonate assay, sulfuric acid/phenol assay, and inductively coupled plasma spectroscopy of gadolinium-DTPA-mannosyl-dextran, respectively. Receptor affinity was measured by Scatchard assay of rabbit liver. Axillary, popliteal, and iliac lymph nodes and each injection site were assayed for radioactivity at 1 and 3 h after injection of approximately 3.7 MBq (0.050 mL) (99m)Tc-DTPA-mannosyl-dextran (0.22 nmol) or filtered (99m)Tc-sulfur colloid into the foot pads. Four animals were studied at each time point. RESULTS: DTPA-mannosyl-dextran had a molecular weight of 35,800 g/mol and a molecular diameter of 7.1 nm. The final amine, mannose, and DTPA densities were 23, 55, and 8 mol per dextran. Labeling yields were in excess of 98% and stable for 6 h. Specific activities of 74 x 10(6) GBq/mol were achieved. The equilibrium dissociation constant for binding to the mannose-terminated glycoprotein receptor was 0.12 +/- 0.07 nmol/L. The popliteal extraction at both 1 h and 3 h was significantly (P < 0.05) higher for (99m)Tc DTPA-mannosyl-dextran (90.1% +/- 10.7% and 97.7% +/- 2.0%, respectively) than for filtered (99m)Tc-sulfur colloid (78.8 +/- 6.5 and 67.4% +/- 26.8%, respectively). (99m)Tc-DTPA-mannosyl-dextran exhibited significantly faster injection site clearance than did filtered (99m)Tc-sulfur colloid. The (99m)Tc-DTPA-mannosyl dextran percentage injected dose (%ID) for the front and rear paws was 52.6 +/- 10.5 and 52.3 +/- 8.0 at 1 h and 45.7 +/- 8.5 and 43.6 +/- 8.2 at 3 h after administration. The filtered (99m)Tc-sulfur colloid %ID for the front and rear paws was 70.4 +/- 11.0 and 66.3 +/- 15.1 at 1 h and 55.5 +/- 7.8 and 66.9 +/- 8.5 at 3 h. Lymph node accumulation of each agent at either 1 or 3 h was not significantly different. CONCLUSION: (99m)Tc-DTPA-mannosyl-dextran is a receptor based sentinel node radiotracer that exhibits the desired properties of rapid injection site clearance and low distal node accumulation. This molecule is the first member of a new class of diagnostic agents based on a macromolecular backbone with a high density of sites for the attachment of substrates and imaging reporters. PMID- 11390563 TI - Positron emission mammography-guided breast biopsy. AB - Positron emission mammography (PEM) is a technique to obtain planar images of the breast for detection of potentially cancerous, radiotracer-avid tumors. To increase the diagnostic accuracy of this method, use of minimally invasive methods (e.g., core biopsy) may be desirable for obtaining tissue samples from lesions detected with PEM. The purpose of this study was to test the capabilities of a novel method for performing PEM-guided stereotactic breast biopsies. METHODS: The PEM system consisted of 2 square (10 x 10 cm) arrays of discrete scintillator crystals. The detectors were mounted on a stereotactic biopsy table. The stereotactic technique used 2 PEM images acquired at +/-15 degrees and a new trigonometric algorithm. The accuracy and precision of the guidance method was tested by placement of small point sources of (18)F at known locations within the field of view of the imager. The calculated positions of the sources were compared with the known locations. In addition, simulated stereotactic biopsies of a breast phantom consisting of a 10-mm-diameter gelatin sphere containing a concentration of (18)F-FDG consistent with that reported for breast cancer were performed. The simulated lesion was embedded in a 4-cm-thick slab of gelatin containing a commonly reported concentration of FDG, simulating a compressed breast (target-to-background ratio, approximately 8.5:1). An anthropomorphic torso phantom was used to simulate tracer uptake in the organs of a patient 1 h after a 370-MBq injection of FDG. Five trials of the biopsy procedure were performed to assess repeatability. Finally, a method for verifying needle positioning was tested. RESULTS: The positions of the point sources were successfully calculated to within 0.6 mm of their true positions with a mean error of +/-0.4 mm. The biopsy procedures, including the method for verification of needle position, were successful in all 5 trials in acquiring samples from the simulated lesions. CONCLUSION: The success of this new technique shows its potential for guiding the biopsy of breast lesions optimally detected with PEM. PMID- 11390564 TI - Radioimmunotherapy of a human lung cancer xenograft with monoclonal antibody RS7: evaluation of (177)Lu and comparison of its efficacy with that of (90)Y and residualizing (131)I. AB - Tumor targeting and therapeutic efficacy of (177)Lu-labeled monoclonal antibody (mAb) RS7 (antiepithelial glycoprotein-1) was evaluated in a human nonsmall cell lung carcinoma xenograft model. The potential of (177)Lu-labeled RS7 was compared with that of RS7 labeled with (90)Y and a residualizing form of (131)I. METHODS: A 1,4,7,10-tetraazacyclododecane-N,N',N",N"'-tetraacetic acid (DOTA) conjugate of RS7 was used for radiolabeling with (177)Lu-acetate or (88/90)Y-acetate. Biodistribution and therapy studies were conducted in nude mice with subcutaneous Calu-3 xenografts. Therapy studies were performed using the maximal tolerated doses (MTDs) of (90)Y-DOTA-RS7 (3.9 MBq [105 microCi]) and (177)Lu-DOTA-RS7 (10.2 MBq [275 microCi]) and compared with the data obtained using the MTD (13.0 MBq [350 microCi]) of a residualizing form of (131)I-RS7. RESULTS: Radiolabeling of RS7-DOTA conjugate with (177)Lu-acetate was facile. (177)Lu-DOTA-RS7 displayed biodistribution results that were nearly identical to that of the (88)Y analog in a paired-label study. The mean percentage injected doses per gram (%ID/g) for (177)Lu-RS7 and (88)Y-RS7 (in parentheses) in tumor were 38.3 %ID/g (39.1 %ID/g), 63.0 %ID/g (66.0 %ID/g), 63.0 %ID/g (65.8 %ID/g), and 34.0 %ID/g (34.9 %ID/g) on days 1, 3, 7, and 14, respectively. Elimination of established tumors, with an initial mean tumor volume of 0.24 cm(3), was shown using doses of (177)Lu-DOTA RS7 ranging from 5.6 to 9.3 MBq (150--250 microCi) per nude mouse, with no significant difference in response rate noted between the doses in this range. Specificity of the therapeutic effect was shown in an isotype-matched control experiment, in which (177)Lu-DOTA-RS7 was markedly more effective than the (177)Lu-DOTA control antibody. A comparison of the therapeutic efficacies of (177)Lu-DOTA-RS7 and (90)Y-DOTA-RS7, using mice with established tumors with an initial mean tumor volume of 0.85 cm(3), indicated similar tumor growth inhibition and similar tumor regrowth profiles. The therapy data were similar to those obtained with residualizing (131)I-RS7 obtained at the same time. CONCLUSION: (177)Lu-RS7 is an effective radioimmunoconjugate for radioimmunotherapy. With its radiophysical properties similar to those of (131)I, coupled with its facile and stable attachment to mAb, (177)Lu promises to be an alternative to (131)I, and a complement to (90)Y, in radioimmunotherapy. PMID- 11390565 TI - Thyroid carcinoma with high levels of function: treatment with (131)I. AB - In some patients with well-differentiated thyroid carcinoma, dosimetry is necessary to avoid toxicity from therapy and to guide prescription of the administered activity of radioiodine. METHODS: The presentations and courses of 2 patients exemplify the points. In the second patient, the clues to the need for dosimetry were the large size of the tumor and high circulating levels of thyroxine in the absence of exogenous hormone. The other patient manifested hyperthyroidism from stimulation of the tumors by thyroid-stimulating immunoglobulin. Dosimetry was performed by published methods. RESULTS: Dosimetry of radioactivity in the body and blood warned of increased irradiation per gigabecquerel of administered (131)I. In each patient, the tumors sequestered a substantial amount of administered (131)I and secreted (131)I-labeled hormones that circulated for days. In 1 patient, the blood time--activity curve was complex, making a broad range of predictions for irradiation to blood and bone marrow. Still, dosimetry gave information that helped to avoid severe toxicity. At, respectively, 1.85 and 2.2 GBq (131)I, initial treatments were relatively low. There was a modest escalation in subsequent administered activities. Leukopenia with neutropenia developed in each patient, and one had moderate thrombocytopenia and anemia, but toxicity appeared to be transient. Each patient had a marked increase in well-being and evidence of reduced tumor function and volume. CONCLUSION: Two patients with advanced, well-differentiated thyroid carcinoma illustrate the need for dosimetry to help prevent toxicity to normal tissues from therapeutic radioiodine. Conversion of radioiodide to circulating radiothyroxine by functioning carcinomas increases the absorbed radiation in normal tissues. Yet, dosimetric data acquired for 4 d or more may be insufficient for accurate calculations of absorbed radiation in blood. Guidelines suggested for avoiding toxicity are based on the circulating thyroxine concentrations, the presence of thyroid stimulators, the amount of radioactivity retained in the body at 48 h, and the general status of the patient. PMID- 11390566 TI - Thyroid stunning after (131)I diagnostic whole-body scanning. PMID- 11390567 TI - The endogenous mammary gland Na(+)/I(-) symporter may mediate effective radioiodide therapy in breast cancer. PMID- 11390568 TI - Empirically supported treatments in pediatric psychology: regimen adherence. AB - OBJECTIVE: To review empirical studies of psychological interventions for nonadherence to medical regimens for three chronic illnesses: asthma, juvenile rheumatoid arthritis (JRA), and type 1 diabetes. METHODS: The Chambless criteria for "promising," "probably efficacious," or "well-established" were applied to 8 intervention studies on asthma, 4 on JRA, and 11 on type 1 diabetes. RESULTS: For asthma, organizational strategies appear probably efficacious in promoting adherence, whereas educational and behavioral strategies appear promising. For JRA, behavioral strategies appear probably efficacious in improving adherence. For type 1 diabetes, multicomponent packages and operant learning procedures appear probably efficacious, whereas cognitive-behavioral strategies appear promising. No interventions were identified as "well-established." CONCLUSIONS: Future studies will need to develop adequate definitions of adherence, accurate methods of assessing adherence, and appropriate designs to evaluate multicomponent treatment programs to advance interventions to the "well established" category. PMID- 11390569 TI - Commentary: pushing the envelope: furthering research on improving adherence to chronic pediatric disease regimens. PMID- 11390570 TI - Commentary: if "an apple a day keeps the doctor away," why is adherence so darn hard? PMID- 11390571 TI - Adolescents' and their friends' health-risk behavior: factors that alter or add to peer influence. AB - OBJECTIVE: To examine models of risk for adolescent health-risk behavior, including family dysfunction, social acceptance, and depression as factors that may compound or mitigate the associations between adolescents' and peers' risk behavior. METHODS: Participants were 527 adolescents in grades 9-12. Adolescents reported on their substance use (cigarette and marijuana use, heavy episodic drinking), violent behavior (weapon carrying, physical fighting), suicidality (suicidal ideation and attempts), and the health-risk behavior of their friends. RESULTS: Adolescents' substance use, violence, and suicidal behavior were related to their friends' substance use, deviance, and suicidal behaviors, respectively. Friends' prosocial behavior was negatively associated with adolescent violence and substance use. Family dysfunction, social acceptance, and depression altered the magnitude of association between peers' and adolescents' risk behavior. In cumulative risk factor models, rates of adolescent health-risk behavior increased twofold with each added risk factor. CONCLUSIONS: Results supported both additive and multiplicative models of risk. Implications for intervention and primary prevention are discussed. PMID- 11390572 TI - Parental distress during pediatric leukemia and posttraumatic stress symptoms (PTSS) after treatment ends. AB - OBJECTIVE: To evaluate prospectively the association between parental anxiety during treatment for childhood leukemia and posttraumatic stress symptoms (PTSS) after treatment ends. A secondary goal is to explore concurrent variables associated with parental avoidance after treatment ends. METHODS: This is a longitudinal follow-up study of 113 parents of children treated for leukemia who previously participated in a study of procedural distress during treatment. Data included parental self-report questionnaires completed during treatment and after treatment. RESULTS: Using hierarchical multiple regression, we found anxiety during treatment to be a significant predictor of later PTSS for mothers, but not fathers. Anxiety, self-efficacy, posttraumatic growth and length of time since treatment ended were associated with parental avoidance. CONCLUSIONS: Highly anxious parents are at risk for PTSS and may benefit from approaches that decrease anxiety during treatment and afterward. Enhancing self-efficacy related to follow-up care and identifying positive aspects of the traumatic experiences are suggested as treatment approaches for families after cancer treatment. PMID- 11390573 TI - Social information processing and magnetic resonance imaging in children with sickle cell disease. AB - OBJECTIVE: To examine social information processing, social skills, and adjustment difficulties in children with sickle cell disease (SCD) as rated by caregivers, teachers, and the children themselves. Children were classified in two groups: cerebral vascular accidents (CVA) (n = 21) or without central nervous system (CNS) pathology (n = 20) on magnetic resonance imaging (MRI). Both groups had HbSS SCD. We compared these two groups and a third group of 11 children who had a milder type of SCD (HbSC). METHODS: Participants referred for evaluation of learning and behavior problems were administered MRIs to ascertain the presence of pathology and a series of measures designed to assess nonverbal emotional decoding abilities and ratings of social emotional functioning. RESULTS: Children with CVA displayed more errors on tasks of facial and vocal emotional decoding than did comparison controls without CVA. CONCLUSIONS: Acquired neurological impairments in children with SCD seemed to be associated with difficulties in the decoding of emotions of other children and adults. We recommend that future research integrate neuropsychological and psychosocial research programs for pediatric chronic illness groups. PMID- 11390575 TI - Initiation of minute virus of mice DNA replication is regulated at the level of origin unwinding by atypical protein kinase C phosphorylation of NS1. AB - Minute virus of mice nonstructural protein NS1 is a multifunctional protein that is involved in many processes necessary for virus propagation. To perform its distinct activities in timely coordinated manner, NS1 was suggested to be regulated by posttranslational modifications, in particular phosphorylation. In fact, NS1 replicative functions are dependent on protein kinase C (PKC) phosphorylation, most likely due to alteration of the biochemical profile of the viral product as determined by comparing native NS1 with its dephosphorylated counterpart. Through the characterization of NS1 mutants at individual PKC consensus phosphorylation sites for their biochemical activities and nickase function, we were able to identify two target atypical PKC phosphorylation sites, T435 and S473, serving as regulatory elements for the initiation of viral DNA replication. Furthermore, by dissociating the energy-dependent helicase activity from the ATPase-independent trans esterification reaction using partially single stranded substrates, we could demonstrate that atypical PKC regulation of NS1 nickase activity occurs at the level of origin unwinding prior to trans esterification. PMID- 11390574 TI - Genetic subtypes, humoral immunity, and human immunodeficiency virus type 1 vaccine development. PMID- 11390576 TI - Genomic features of intertypic recombinant sabin poliovirus strains excreted by primary vaccinees. AB - The trivalent oral poliomyelitis vaccine (OPV) contains three different poliovirus serotypes. It use therefore creates particularly favorable conditions for mixed infection of gut cells, and indeed intertypic vaccine-derived recombinants (VdRec) have been frequently found in patients with vaccine associated paralytic poliomyelitis. Nevertheless, there have not been extensive searches for VdRec in healthy vaccinees following immunization with OPV. To determine the incidence of VdRec and their excretion kinetics in primary vaccinees, and to establish the general genomic features of the corresponding recombinant genomes, we characterized poliovirus isolates excreted by vaccinees following primary immunization with OPV. Isolates were collected from 67 children 2 to 60 days following vaccination. Recombinant strains were identified by multiple restriction fragment length polymorphism assays. The localization of junction sites in recombinant genomes was also determined. VdRec excreted by vaccinees were first detected 2 to 4 days after vaccination. The highest rate of recombinants was on day 14. The frequency of VdRec depends strongly on the serotype of the analyzed isolates (2, 53, and 79% of recombinant strains in the last-excreted type 1, 2, and 3 isolates, respectively). Particular associations of genomic segments were preferred in the recombinant genomes, and recombination junctions were found in the genomic region encoding the nonstructural proteins. Recombination junctions generally clustered in particular subgenomic regions that were dependent on the serotype of the isolate and/or on the associations of genomic segments in recombinants. Thus, VdRec are frequently excreted by vaccinees, and the poliovirus replication machinery requirements or selection factors appear to act in vivo to shape the features of the recombinant genomes. PMID- 11390577 TI - Vaccinia virus A30L protein is required for association of viral membranes with dense viroplasm to form immature virions. AB - The previously uncharacterized A30L gene of vaccinia virus has orthologs in all vertebrate poxviruses but no recognizable nonpoxvirus homologs or functional motifs. We determined that the A30L gene was regulated by a late promoter and encoded a protein of approximately 9 kDa. Immunoelectron microscopy of infected cells indicated that the A30L protein was associated with viroplasm enclosed by crescent and immature virion membranes. The A30L protein was also present in mature virions and was partially released by treatment with a nonionic detergent and reducing agent, consistent with a location in the matrix between the core and envelope. To determine the role of the A30L protein, we constructed a stringent conditional lethal mutant with an inducible A30L gene. In the absence of inducer, synthesis of viral early and late proteins occurred but the proteolytic processing of certain core proteins was inhibited, suggesting an assembly block. Inhibition of virus maturation was confirmed by electron microscopy. Under nonpermissive conditions, we observed aberrant large, dense, granular masses of viroplasm with clearly defined margins; viral crescent membranes that appeared normal except for their location at a distance from viroplasm; empty immature virions; and an absence of mature virions. The data indicated that the A30L protein is needed for vaccinia virus morphogenesis, specifically the association of the dense viroplasm with viral membranes. PMID- 11390578 TI - A particle-associated glycoprotein signal peptide essential for virus maturation and infectivity. AB - Signal peptides (SP) are key determinants for targeting glycoproteins to the secretory pathway. Here we describe the involvement in particle maturation as an additional function of a viral glycoprotein SP. The SP of foamy virus (FV) envelope glycoprotein is predicted to be unusually long. Using an SP-specific antiserum, we demonstrate that its proteolytic removal occurs posttranslationally by a cellular protease and that the major N-terminal cleavage product, gp18, is found in purified viral particles. Analysis of mutants in proposed signal peptidase cleavage positions and N-glycosylation sites revealed an SP about 148 amino acids (aa) in length. FV particle release from infected cells requires the presence of cognate envelope protein and cleavage of its SP sequence. An N terminal 15-aa SP domain with two conserved tryptophan residues was found to be essential for the egress of FV particles. While the SP N terminus was found to mediate the specificity of FV Env to interact with FV capsids, it was dispensable for Env targeting to the secretory pathway and FV envelope-mediated infectivity of murine leukemia virus pseudotypes. PMID- 11390579 TI - Exploitation of the low fidelity of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase and the nucleotide composition bias in the HIV-1 genome to alter the drug resistance development of HIV. AB - The RNA genome of the lentivirus human immunodeficiency virus type 1 (HIV-1) is significantly richer in adenine nucleotides than the statistically equal distribution of the four different nucleotides that is expected. This compositional bias may be due to the guanine-to-adenine (G-->A) nucleotide hypermutation of the HIV genome, which has been explained by dCTP pool imbalances during reverse transcription. The adenine nucleotide bias together with the poor fidelity of HIV-1 reverse transcriptase markedly enhances the genetic variation of HIV and may be responsible for the rapid emergence of drug-resistant HIV-1 strains. We have now attempted to counteract the normal mutational pattern of HIV 1 in response to anti-HIV-1 drugs by altering the endogenous deoxynucleoside triphosphate pool ratios with antimetabolites in virus-infected cell cultures. We showed that administration of these antimetabolic compounds resulted in an altered drug resistance pattern due to the reversal of the predominant mutational flow of HIV (G-->A) to an adenine-to-guanine (A-->G) nucleotide pattern in the intact HIV-1-infected lymphocyte cultures. Forcing the virus to change its inherent nucleotide bias may lead to better control of viral drug resistance development. PMID- 11390580 TI - The major core protein P4a (A10L gene) of vaccinia virus is essential for correct assembly of viral DNA into the nucleoprotein complex to form immature viral particles. AB - The vaccinia virus (VV) A10L gene codes for a major core protein, P4a. This polypeptide is synthesized at late times during viral infection and is proteolytically cleaved during virion assembly. To investigate the role of P4a in the virus life cycle and morphogenesis, we have generated an inducer-dependent conditional mutant (VVindA10L) in which expression of the A10L gene is under the control of the Escherichia coli lacI operator/repressor system. Repression of the A10L gene severely impairs virus growth, as observed by both the inability of the virus to form plaques and the 2-log reduction of viral yields. This defect can be partially overcome by addition of the inducer isopropyl-beta-D thiogalactopyranoside (IPTG). Synthesis of viral proteins other than P4a occurred, although early shutoff of host protein synthesis and expression of viral late polypeptides are clearly delayed, both in the absence and in the presence of IPTG, compared with cells infected with the parental virus. Viral DNA replication and concatemer resolution appeared to proceed normally in the absence of the A10L gene product. In cells infected with VVindA10L in the absence of the inducer virion assembly is blocked, as defined by electron microscopy. Numerous spherical immature viral particles that appear devoid of dense viroplasmic material together with highly electron-dense regular structures are abundant in VVindA10L-infected cells. These regularly spaced structures can be specifically labeled with anti-DNA antibodies as well as with a DNase-gold conjugate, indicating that they contain DNA. Some images suggest that these DNA structures enter into spherical immature viral particles. In this regard, although it has not been firmly established, it has been suggested that DNA uptake occurs after formation of spherical immature particles. Overall, our results showed that P4a and/or its cleaved products are essential for the correct assembly of the nucleoprotein complex within immature viral particles. PMID- 11390581 TI - Promoter-proximal regulatory elements involved in oriP-EBNA1-independent and dependent activation of the Epstein-Barr virus C promoter in B-lymphoid cell lines. AB - The identification of the cellular factors that control the transcription regulatory activity of the Epstein-Barr virus C promoter (Cp) is fundamental to the understanding of the molecular mechanisms that control virus latent gene expression. Using transient transfection of reporter plasmids in group I phenotype B-lymphoid cells, we have previously shown that the -248 to -55 region (-248/-55 region) of Cp contains elements that are essential for oriPI-EBNA1 dependent as well as oriPI-EBNA1-independent activation of the promoter. We now establish the importance of this region by a detailed mutational analysis of reporter plasmids carrying Cp regulatory sequences together with or without oriPI. The reporter plasmids were transfected into group I phenotype Rael cells and group III phenotype cbc-Rael cells, and the Cp activity measured was correlated with the binding of candidate transcription factors in electrophoretic mobility shift assays and further assessed in cotransfection experiments. We show that the NF-Y transcription factor interacts with the previously identified CCAAT box in the -71/-63 Cp region (M. T. Puglielli, M. Woisetschlaeger, and S. H. Speck, J. Virol. 70:5758-5768, 1996). We also show that members of the C/EBP transcription factor family interact with a C/EBP consensus sequence in the -119/ 112 region of Cp and that this interaction is important for promoter activity. A central finding is the identification of a GC-rich sequence in the -99/-91 Cp region that is essential for oriPI-EBNA1-independent as well as oriPI-EBNA1 dependent activity of the promoter. This region contains overlapping binding sites for Sp1 and Egr-1, and our results suggest that Sp1 is a positive and Egr-1 is a negative regulator of Cp activity. Furthermore, we demonstrate that a reporter plasmid that in addition to oriPI contains only the -111/+76 region of Cp still retains the ability to be activated by EBNA1. PMID- 11390582 TI - Human immunodeficiency virus type 1 (HIV-1)-induced GRO-alpha production stimulates HIV-1 replication in macrophages and T lymphocytes. AB - We examined the early effects of infection by CCR5-using (R5 human immunodeficiency virus [HIV]) and CXCR4-using (X4 HIV) strains of HIV type 1 (HIV 1) on chemokine production by primary human monocyte-derived macrophages (MDM). While R5 HIV, but not X4 HIV, replicated in MDM, we found that the production of the C-X-C chemokine growth-regulated oncogene alpha (GRO-alpha) was markedly stimulated by X4 HIV and, to a much lesser extent, by R5 HIV. HIV-1 gp120 engagement of CXCR4 initiated the stimulation of GRO-alpha production, an effect blocked by antibodies to CXCR4. GRO-alpha then fed back and stimulated HIV-1 replication in both MDM and lymphocytes, and antibodies that neutralize GRO-alpha or CXCR2 (the receptor for GRO-alpha) markedly reduced viral replication in MDM and peripheral blood mononuclear cells. Therefore, activation of MDM by HIV-1 gp120 engagement of CXCR4 initiates an autocrine-paracrine loop that may be important in disease progression after the emergence of X4 HIV. PMID- 11390583 TI - Potential sources of the 1995 Venezuelan equine encephalitis subtype IC epidemic. AB - Venezuelan equine encephalitis viruses (VEEV) belonging to subtype IC have caused three (1962-1964, 1992-1993 and 1995) major equine epizootics and epidemics. Previous sequence analyses of a portion of the envelope glycoprotein gene demonstrated a high degree of conservation among isolates from the 1962-1964 and the 1995 outbreaks, as well as a 1983 interepizootic mosquito isolate from Panaquire, Venezuela. However, unlike subtype IAB VEEV that were used to prepare inactivated vaccines that probably initiated several outbreaks, subtype IC viruses have not been used for vaccine production and their conservation cannot be explained in this way. To characterize further subtype IC VEEV conservation and to evaluate potential sources of the 1995 outbreak, we sequenced the complete genomes of three isolates from the 1962-1964 outbreak, the 1983 Panaquire interepizootic isolate, and two isolates from 1995. The sequence of the Panaquire isolate, and that of virus isolated from a mouse brain antigen prepared from subtype IC strain P676 and used in the same laboratory, suggested that the Panaquire isolate represents a laboratory contaminant. Some authentic epizootic IC strains isolated 32 years apart showed a greater degree of sequence identity than did isolates from the same (1962-1964 or 1995) outbreak. If these viruses were circulating and replicating between 1964 and 1995, their rate of sequence evolution was at least 10-fold lower than that estimated during outbreaks or that of closely related enzootic VEEV strains that circulate continuously. Current understanding of alphavirus evolution is inconsistent with this conservation. This subtype IC VEEV conservation, combined with phylogenetic relationships, suggests the possibility that the 1995 outbreak was initiated by a laboratory strain. PMID- 11390584 TI - Thymic lesions in cats infected with a pathogenic molecular clone or an ORF-A/2 deficient molecular clone of feline immunodeficiency virus. AB - Previous studies using feline immunodeficiency virus (FIV) molecular clones lacking the putative transactivator gene (ORF-A/2) failed to address the issue of thymus pathogenesis or investigate the levels of viral replication in separate lymphoid compartments (Y. Inoshima, et al., J. Virol. 70:8518-8526, 1996; E. E. Sparger, et al., Virology 205:546-553, 1994). Using a highly pathogenic molecular clone of FIV, JSY3, and an ORF-A/2-deficient mutant, JSY3DeltaORF-A/2, we compared viral replication and the extent of thymic dysfunction as measured by the formation of lymphoid follicles and alteration of the thymocyte subsets. Viral replication was reduced in JSY3DeltaORF-A/2-infected cats as measured by lymphocyte coculture, immunohistochemistry, and quantitative PCR. Cell-associated viral load measured by lymphocyte coculture varied in a tissue-dependent manner with replication highest in lymphocytes isolated from the thymus, lower in those from the peripheral blood, and lowest in those from lymph node. Thymic proviral load and the number of viral p24 Gag-positive cells within the thymus detected by immunohistochemistry were also reduced. In addition, the onset of a reduced peripheral blood CD4/CD8 ratio was delayed in JSY3DeltaORF-A/2-infected cats. The formation and extent of thymic lymphoid follicular hyperplasia were similar in JSY3 and JSY3DeltaORF-A/2-infected cats as measured by anticytokeratin immunohistochemistry and flow cytometry for percent pan T-negative, immunoglobulin G-positive cells within the thymus. In contrast, comparison of thymocyte subpopulations demonstrated a reduced expansion of single-positive CD4( ) CD8(+) thymocytes in JSY3DeltaORF-A/2-infected cats. Level of viral replication, therefore, may not correlate with the formation of thymic lymphoid follicles but may correlate with the expansion of the single-positive CD4(-) CD8(+) thymocyte subpopulation. PMID- 11390585 TI - Morbilliviruses use signaling lymphocyte activation molecules (CD150) as cellular receptors. AB - Morbilliviruses comprise measles virus, canine distemper virus, rinderpest virus, and several other viruses that cause devastating human and animal diseases accompanied by severe immunosuppression and lymphopenia. Recently, we have shown that human signaling lymphocyte activation molecule (SLAM) is a cellular receptor for measles virus. In this study, we examined whether canine distemper and rinderpest viruses also use canine and bovine SLAMs, respectively, as cellular receptors. The Onderstepoort vaccine strain and two B95a (marmoset B cell line) isolated strains of canine distemper virus caused extensive cytopathic effects in normally resistant CHO (Chinese hamster ovary) cells after expression of canine SLAM. The Ako vaccine strain of rinderpest virus produced strong cytopathic effects in bovine SLAM-expressing CHO cells. The data on entry with vesicular stomatitis virus pseudotypes bearing measles, canine distemper, or rinderpest virus envelope proteins were consistent with development of cytopathic effects in SLAM-expressing CHO cell clones after infection with the respective viruses, confirming that SLAM acts at the virus entry step (as a cellular receptor). Furthermore, most measles, canine distemper, and rinderpest virus strains examined could any use of the human, canine, and bovine SLAMs to infect cells. Our findings suggest that the use of SLAM as a cellular receptor may be a property common to most, if not all, morbilliviruses and explain the lymphotropism and immunosuppressive nature of morbilliviruses. PMID- 11390586 TI - Nef enhances human immunodeficiency virus type 1 infectivity resulting from intervirion fusion: evidence supporting a role for Nef at the virion envelope. AB - The human immunodeficiency virus type 1 (HIV-1) accessory protein Nef stimulates viral infectivity by facilitating an early event in the HIV-1 life cycle. Although no structural or biochemical defects in Nef-defective HIV-1 particles have been demonstrated, the Nef protein is incorporated into HIV-1 particles. To localize the function of Nef within the virus particle, we developed a novel technology involving fusion of enveloped donor HIV-1 particles bearing core defects with envelope-defective target virions bearing HIV-1 receptors. Although neither virus alone was capable of infecting CD4(+) target cells, the incubation of donor and target virions prior to addition to target cells resulted in infection. This effect, termed "virion transcomplementation," required a functional Env protein on the donor virus and CD4 and an appropriate coreceptor on target virions. To provide evidence for intervirion fusion as the mechanism of complementation, experiments were performed using dual-enveloped HIV-1 particles bearing both HIV-1 and ecotropic murine leukemia virus (E-MLV) Env proteins as donor virions. Infection of CD4-negative target cells bearing E-MLV receptors was prevented by HIV-1 entry inhibitors when added before, but not after, incubation of donor and target virions prior to the addition to cells. When we used Nef(+) and Nef(-) donor and target virions, Nef enhanced infection when present in donor virions. In contrast, no effect of Nef was detected when present in the target virus. These results reveal a potential mechanism for enhancing HIV-1 diversity in vivo through the rescue of defective viral genomes and provide a novel genetic system for the functional analysis of virion-associated proteins in HIV-1 infection. PMID- 11390587 TI - Cytokine production by Vgamma(+)-T-cell subsets is an important factor determining CD4(+)-Th-cell phenotype and susceptibility of BALB/c mice to coxsackievirus B3-induced myocarditis. AB - Two coxsackievirus B3 (CVB3) variants (H3 and H310A1) differ by a single amino acid mutation in the VP2 capsid protein. H3 induces severe myocarditis in BALB/c mice, but H310A1 is amyocarditic. Infection with H3, but not H310A1, preferentially activates Vgamma4 Vdelta4 cells, which are strongly positive for gamma interferon (IFN-gamma), whereas Vgamma1 Vdelta4 cells are increased in both H3 and H310A1 virus-infected animals. Depletion of Vgamma1(+) cells using monoclonal anti-Vgamma1 antibody enhanced myocarditis and CD4(+)-, IFN-gamma(+) cell responses in both H3- and H310A1-infected mice yet decreased the CD4(+)-, IL 4(+)-cell response. Depleting Vgamma4(+) cells suppressed myocarditis and reduced CD4(+) IFN-gamma(+) cells but increased CD4(+) IL-4(+) T cells. The role of cytokine production by Vgamma1(+) and Vgamma4(+) T cells was investigated by adoptively transferring these cells isolated from H3-infected BALB/c Stat4 knockout (Stat4ko) (defective in IFN-gamma expression) or BALB/c Stat6ko (defective in IL-4 expression) mice into H3 virus-infected wild-type BALB/c recipients. Vgamma4 and Vgamma1(+) T cells from Stat4ko mice expressed IL-4 but no or minimal IFN-gamma, whereas these cell populations derived from Stat6ko mice expressed IFN-gamma but no IL-4. Stat4ko Vgamma1(+) cells (IL-4(+)) suppress myocarditis. Stat6ko Vgamma1(+) cells (IFN-gamma(+)) were not inhibitory. Stat6ko Vgamma4(+) cells (IFN-gamma(+)) significantly enhanced myocarditis. Stat4ko Vgamma4(+) cells (IL-4(+)) neither inhibited nor enhanced disease. These results show that distinct gammadelta-T-cell subsets control myocarditis susceptibility and bias the CD4(+)-Th-cell response. The cytokines produced by the Vgamma subpopulation have a significant influence on the CD4(+)-Th-cell phenotype. PMID- 11390588 TI - Cleavage in and around the DR1 element of the A sequence of herpes simplex virus type 1 relevant to the excision of DNA fragments with length corresponding to one and two units of the A sequence. AB - The A sequence of herpes simplex virus type 1 (HSV-1) is a region bracketed by two direct repeats named DR1. Concatemeric HSV-1 DNA, the product of DNA replication, is cleaved at a specific site on the second DR1 distal from the S component (authentic cleavage) to yield unit-length linear HSV-1 DNA prior to or during packaging of HSV-1 DNA. The presence of two DNA bands, of 0.25 kb (shorter band) and 0.5 kb (longer band), the lengths of which correspond to one and two units of the A sequence, was identified using acrylamide gel electrophoresis of HSV-1 DNA preparations extracted by the method of Hirt. Twelve DNA fragments from each band were molecularly cloned, and nucleotide sequences were determined. Both termini of eight (67%) DNA clones from the shorter band corresponded to the specific cleavage site on DR1. Five (41%) DNA clones from the longer band had a terminus corresponding to the specific cleavage site on DR1 on one side, but not on the opposite side. Thirteen (54%) of 24 termini of 12 analyzed DNA clones from the longer band were in and around DR1. Thus, cleavage events of DR1 can be classified into three categories: (i) authentic cleavage; (ii) site-specific cleavage on the third DR1 distal from the S component (secondary site-specific cleavage), which is related to the generation of the shorter DNA band in combination with authentic cleavage; and (iii) less-specific cleavage events in and around other DR1 elements which relate to the generation of the longer DNA band. PMID- 11390590 TI - Immunoreceptor tyrosine-based activation motif-dependent signaling by Kaposi's sarcoma-associated herpesvirus K1 protein: effects on lytic viral replication. AB - The Kaposi's sarcoma-associated herpesvirus (KSHV) K1 gene encodes a polypeptide bearing an immunoreceptor tyrosine-based activation motif (ITAM) that is constitutively active for ITAM-based signal transduction. Although ectopic overexpression of K1 in cultured fibroblasts can lead to growth transformation, in vivo this gene is primarily expressed in lymphoid cells undergoing lytic infection. Here we have examined function of K1 in the setting of lytic replication, through the study of K1 mutants lacking functional ITAMs. Expression of such mutants in BJAB cells cotransfected with wild-type K1 results in dramatic inhibition of K1 signal transduction, as judged by impaired activation of Syk kinase and phospholipase C-gamma2 as well as by diminished expression of a luciferase reporter gene dependent upon K1-induced calcium and Ras signaling. Thus, the mutants behave as dominantly acting inhibitors of K1 function. To assess the role of K1 in lytic replication, we introduced these K1 mutants into BCBL-1 cells, a B-cell lymphoma line latently infected with KSHV, and induced lytic replication by ectopic expression of the KSHV ORF50 transactivator. Expression of lytic cycle genes was diminished up to 80% in the presence of a K1 dominant negative mutant. These inhibitory effects could be overridden by tetradecanoyl phorbol acetate treatment, indicating that inhibition was not due to irreversible cell injury and suggesting that other signaling events could bypass the block. We conclude that ITAM-dependent signaling by K1 is not absolutely required for lytic reactivation but functions to modestly augment lytic replication in B cells, the natural reservoir of KSHV. PMID- 11390589 TI - Induction of neutralizing antibodies and gag-specific cellular immune responses to an R5 primary isolate of human immunodeficiency virus type 1 in rhesus macaques. AB - The ability to generate antibodies that cross-neutralize diverse primary isolates is an important goal for human immunodeficiency virus type 1 (HIV-1) vaccine development. Most of the candidate HIV-1 vaccines tested in humans and nonhuman primates have failed in this regard. Past efforts have focused almost entirely on the envelope glycoproteins of a small number of T-cell line-adapted strains of the virus as immunogens. Here we assessed the immunogenicity of noninfectious virus-like particles (VLP) consisting of Gag, Pro (protease), and Env from R5 primary isolate HIV-1(Bx08). Immunogens were delivered to rhesus macaques in the form of either purified VLP, recombinant DNA and canarypox (ALVAC) vectors engineered to express VLP, or a combination of these products. Seroconversion to Gag and Pro was detected in all of the immunized animals. Antibodies that could neutralize HIV-1(Bx08) were detected in animals that received (i) coinoculations with DNA(Bx08) and VLP(Bx08), (ii) DNA(Bx08) followed by ALVAC(Bx08) boosting, and (iii) VLP(Bx08) alone. The neutralizing antibodies were highly strain specific despite the fact that they did not appear to be directed to linear epitopes in the V3 loop. Virus-specific cellular immune responses also were generated, as judged by the presence of Gag-specific gamma interferon (IFN-gamma) producing cells. These cellular immune responses required the inclusion of DNA(Bx08) in the immunization modality, since few or no IFN-gamma-producing cells were detected in animals that received either VLP(Bx08) or ALVAC(Bx08) alone. The results demonstrate the feasibility of generating neutralizing antibodies and cellular immune responses that target an R5 primary HIV-1 isolate by vaccination in primates. PMID- 11390591 TI - Notch1IC partially replaces EBNA2 function in B cells immortalized by Epstein Barr virus. AB - Immortalization of B cells by Epstein-Barr virus (EBV) depends on the virally encoded EBNA2 protein. Although not related by sequence, the cellular Notch protein and EBNA2 share several biochemical and functional properties, such as interaction with CBF1 and the ability to activate transcription of a number of cellular and viral genes. Whether these similarities are coincidental or exemplify EBNA2 mimicry of evolutionarily conserved cellular signaling pathways is unclear. We therefore investigated whether activated forms of Notch could substitute for EBNA2 in maintaining the immortalized phenotype of EBV-infected B cells. To address this question, we devised a transcomplementation system using EREB2.5 cells. EREB2.5 cells are immortalized by EBV expressing a conditional estrogen receptor EBNA2 fusion protein (EREBNA2), and cellular proliferation is dependent on the availability of estrogen. Withdrawal of estrogen results in inactivation of EREBNA2, leading to growth arrest and eventually to cell death. Transduction of EREB2.5 cells with a lentiviral vector expressing wild-type EBNA2 rescued EREB2.5 cells from the growth-inhibitory effects of estrogen deprivation, in contrast to transduction with the lentivirus vector alone. EREB2.5 cells were also rescued by enforced expression of human Notch1IC after estrogen starvation, but this effect was restricted to cells expressing high levels of the transcription factor. Compared to wild-type EBNA2-expressing EREB2.5 cells, the Notch-expressing cells expanded more slowly after estrogen starvation, and once established, they continued to display a lower proliferation rate. Analysis of viral and cellular gene expression from transduced EREB2.5 cells after estrogen withdrawal indicated that both wild-type EBNA2- and Notch1IC-positive cells expressed c-Myc at levels similar to those found in parental EREB2.5 cells. However, the latter cells expressed LMP-1 far less efficiently than cells transduced with the wild-type EBNA2 gene. Cells rescued by either wild-type EBNA2 or Notch1IC expressed surface CD21 and CD23 proteins, but not CD10, indicating that induction of relevant type III latency markers was maintained. The data imply that both Notch and EBNA2 activate an important subset of cellular genes associated with type III latency and B-cell growth, while EBNA2 more efficiently induces important viral genes, such as LMP-1. Thus, exploitation of conserved Notch-related signaling pathways may represent a key mechanism by which EBNA2 contributes to EBV-induced cell immortalization. PMID- 11390592 TI - Adenovirus vectors with the 100K gene deleted and their potential for multiple gene therapy applications. AB - The 100K protein has a number of critical roles vital for successful completion of the late phases of the adenovirus (Ad) life cycle. We hypothesized that the introduction of deletions within the 100K gene would allow for the production of a series of new classes of Ad vector, including one that is replication competent but blocked in the ability to carry out many late-phase Ad functions. Such a vector would have potential for several gene therapy applications, based upon its ability to increase the copy number of the transgene encoded by the vector (via genome replication) while decreasing the side effects associated with Ad late gene expression. To efficiently produce 100K-deleted Ad ([100K-]Ad) vectors, an E1- and 100K-complementing cell line (K-16) was successfully isolated. Transfection of an [E1-,100K-]Ad vector genome into the K-16 cells readily yielded high titers of the vector. After infection of noncomplementing cells, we demonstrated that [100K-]Ad vectors have a significantly decreased ability to express several Ad late genes. Additionally, if the E1 gene was present in the infected noncomplementing cells, [100K-]Ad vectors were capable of replicating their genomes to high copy number, but were significantly blocked in their ability to efficiently encapsidate the replicated genomes. Injection of an [E1 ,100K-]Ad vector in vivo also correlated with significantly decreased hepatotoxicity, as well as prolonged vector persistence. In summary, the unique properties of [100K-]Ad vectors suggest that they may have utility in a variety of gene therapy applications. PMID- 11390593 TI - Human lymphocyte apoptosis after exposure to influenza A virus. AB - Infection of humans with influenza A virus (IAV) results in a severe transient leukopenia. The goal of these studies was to analyze possible mechanisms behind this IAV-induced leukopenia with emphasis on the potential induction of apoptosis of lymphocytes by the virus. Analysis of lymphocyte subpopulations after exposure to IAV showed that a portion of CD3(+), CD4(+), CD8(+), and CD19(+) lymphocytes became apoptotic (terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling positive). The percentage of cells that are infected was shown to be less than the percentage of apoptotic cells, suggesting that direct effects of cell infection by the virus cannot account fully for the high level of cell death. Removal of monocytes-macrophages after IAV exposure reduced the percent of lymphocytes that were apoptotic. Treatment of virus-exposed cultures with anti tumor necrosis factor alpha did not reduce the percentage of lymphocytes that were apoptotic. In virus-exposed cultures treated with anti-FasL antibody, recombinant soluble human Fas, Ac-DEVD-CHO (caspase-3 inhibitor), or Z-VAD-FMK (general caspase inhibitor), apoptosis and production of the active form of caspase-3 was reduced. The apoptotic cells were Fas-high-density cells while the nonapoptotic cells expressed a low density of Fas. The present studies showed that Fas-FasL signaling plays a major role in the induction of apoptosis in lymphocytes after exposure to IAV. Since the host response to influenza virus commonly results in recovery from the infection, with residual disease uncommon, lymphocyte apoptosis likely represents a part of an overall beneficial immune response but could be a possible mechanism of disease pathogenesis. PMID- 11390594 TI - Theiler's murine encephalomyelitis virus induces apoptosis in gamma interferon activated M1 differentiated myelomonocytic cells through a mechanism involving tumor necrosis factor alpha (TNF-alpha) and TNF-alpha-related apoptosis-inducing ligand. AB - Infection of susceptible mice with the low-neurovirulence Theiler's murine encephalomyelitis virus strain BeAn results in an inflammatory demyelinating disease similar to multiple sclerosis. While the majority of virus antigen is detected in central nervous system macrophages (Mphis), few infiltrating Mphis are infected. We used the myelomonocytic precursor M1 cell line to study BeAn virus-Mphi interactions in vitro to elucidate mechanisms for restricted virus expression. We have shown that restricted BeAn infection of M1 cells differentiated in vitro (M1-D) results in apoptosis. In this study, BeAn infection of gamma interferon (IFN-gamma)-activated M1-D cells also resulted in apoptosis but with no evidence of virus replication or protein expression. RNase protection assays of M1-D cellular RNA revealed up-regulation of Fas and the p55 chain of the tumor necrosis factor alpha (TNF-alpha) receptor transcripts with IFN-gamma activation. BeAn infection of activated cells resulted in increased caspase 8 mRNA transcripts and the appearance of TNF-alpha-related apoptosis inducing ligand (TRAIL) 4 h postinfection. Both unactivated and activated M1-D cells expressed TRAIL receptors (R1 and R2), but only activated cells were killed by soluble TRAIL. Activated cells were also susceptible to soluble FasL- and TNF alpha-induced apoptosis. The data suggest that IFN-gamma-activated M1-D cell death receptors become susceptible to their ligands and that the cells respond to BeAn virus infection by producing the ligands TNF-alpha and TRAIL to kill the susceptible cells. Unactivated cells are not susceptible to FasL or TRAIL and require virus replication to initiate apoptosis. Therefore, two mechanisms of apoptosis induction can be triggered by BeAn infection: an intrinsic pathway requiring virus replication and an extrinsic pathway signaling through the death receptors. PMID- 11390595 TI - Immunogenicity and protective efficacy of recombinant human T-cell leukemia/lymphoma virus type 1 NYVAC and naked DNA vaccine candidates in squirrel monkeys (Saimiri sciureus). AB - We assessed the immunogenicities and efficacies of two highly attenuated vaccinia virus-derived NYVAC vaccine candidates encoding the human T-cell leukemia/lymphoma virus type 1 (HTLV-1) env gene or both the env and gag genes in prime-boost pilot regimens in combination with naked DNA expressing the HTLV-1 envelope. Three inoculations of NYVAC HTLV-1 env at 0, 1, and 3 months followed by a single inoculation of DNA env at 9 months protected against intravenous challenge with HTLV-1-infected cells in one of three immunized squirrel monkeys. Furthermore, humoral and cell-mediated immune responses against HTLV-1 Env could be detected in this protected animal. However, priming the animal with a single dose of env DNA, followed by immunization with the NYVAC HTLV-1 gag and env vaccine at 6, 7, and 8 months, protected all three animals against challenge with HTLV-1-infected cells. With this protocol, antibodies against HTLV-1 Env and cell mediated responses against Env and Gag could also be detected in the protected animals. Although the relative superiority of a DNA prime-NYVAC boost regimen over addition of the Gag component as an immunogen cannot be assessed directly, our findings nevertheless show that an HTLV-1 vaccine approach is feasible and deserves further study. PMID- 11390596 TI - Human cytomegalovirus chemokine receptor gene US28 is transcribed in latently infected THP-1 monocytes. AB - The human cytomegalovirus (HCMV) US28 gene product, pUS28, is a G protein-coupled receptor that interacts with both CC and CX(3)C chemokines. To date, the role of pUS28 in immune evasion and cell migration has been studied only in cell types that can establish productive HCMV infection. We show that HCMV can latently infect THP-1 monocytes and that during latency US28 is transcribed. We also show that the transcription is sustained during differentiation of the THP-1 monocytes. Since cells expressing pUS28 were previously shown to adhere to immobilized CX(3)C chemokines (C. A. Haskell, M. D. Cleary, and I. F. Charo, J. Biol. Chem. 275:34183-34189, 2000), we hypothesize that latently infected circulating monocytes express pUS28, thereby enabling adhesion of these cells to CX(3)C-exposing endothelium. Consequently, the US28-encoded chemokine receptor may play an important role in dissemination of latent HCMV. PMID- 11390597 TI - Immature monocyte-derived dendritic cells are productively infected with herpes simplex virus type 1. AB - Herpes simplex viruses (HSV) have developed several immunoevasive strategies. Here we demonstrate a novel mechanism by which HSV type 1 may interfere with the immune response through infection of immature dendritic cells (DC) and selective downmodulation of costimulatory molecules. In our study we show productive infection of immature monocyte-derived DC, which closely resemble sessile Langerhans cells, by sequential expression of immediate-early, early, and late viral proteins and of glycoprotein D mRNA, as well as production of infectious virus of moderate titers. Infection was cytopathic, with the progressive loss of 20 to 45% of cells from 24 to 48 h after infection, with no more than 80% of DC found to be infected. These results are in contrast to those of previous findings of nonpermissive or abortive infection of monocytes and mature monocyte-derived DC. Infection of immature DC also led to selective and asynchronous downregulation of CD1a, CD40, CD54 (ICAM-1) (12 h postinfection), CD80 (24 h postinfection), and CD86 (48 h postinfection) but not of CD11c or major histocompatibility complex class I and II molecules when compared to DC exposed to UV-inactivated virus. Thus, we propose that productive infection of epidermal Langerhans cells in vivo may lead to delayed activation of T cells, allowing more time for replication of HSV type 1 in epidermal cells. PMID- 11390598 TI - Attrition of bystander CD8 T cells during virus-induced T-cell and interferon responses. AB - Experiments designed to distinguish virus-specific from non-virus-specific T cells showed that bystander T cells underwent apoptosis and substantial attrition in the wake of a strong T-cell response. Memory CD8 T cells (CD8(+) CD44(hi)) were most affected. During acute viral infection, transgenic T cells that were clearly defined as non-virus specific decreased in number and showed an increase in apoptosis. Also, use of lymphocytic choriomeningitis virus (LCMV) carrier mice, which lack LCMV-specific T cells, showed a significant decline in non-virus specific memory CD8 T cells that correlated to an increase in apoptosis in response to the proliferation of adoptively transferred virus-specific T cells. Attrition of T cells early during infection correlated with the alpha/beta interferon (IFN-alpha/beta) peak, and the IFN inducer poly(I:C) caused apoptosis and attrition of CD8(+) CD44(hi) T cells in normal mice but not in IFN-alpha/beta receptor-deficient mice. Apoptotic attrition of bystander T cells may make room for the antigen-specific expansion of T cells during infection and may, in part, account for the loss of T-cell memory that occurs when the host undergoes subsequent infections. PMID- 11390599 TI - Markedly increased susceptibility to natural sheep scrapie of transgenic mice expressing ovine prp. AB - The susceptibility of sheep to scrapie is known to involve, as a major determinant, the nature of the prion protein (PrP) allele, with the VRQ allele conferring the highest susceptibility to the disease. Transgenic mice expressing in their brains three different ovine PrP(VRQ)-encoding transgenes under an endogenous PrP-deficient genetic background were established. Nine transgenic (tgOv) lines were selected and challenged with two scrapie field isolates derived from VRQ-homozygous affected sheep. All inoculated mice developed neurological signs associated with a transmissible spongiform encephalopathy (TSE) disease and accumulated a protease-resistant form of PrP (PrPres) in their brains. The incubation duration appeared to be inversely related to the PrP steady-state level in the brain, irrespective of the transgene construct. The survival time for animals from the line expressing the highest level of PrP was reduced by at least 1 year compared to those of two groups of conventional mice. With one isolate, the duration of incubation was as short as 2 months, which is comparable to that observed for the rodent TSE models with the briefest survival times. No survival time reduction was observed upon subpassaging of either isolate, suggesting no need for adaptation of the agent to its new host. Overexpression of the transgene was found not to be required for transmission to be accelerated compared to that observed with wild-type mice. Conversely, transgenic mice overexpressing murine PrP were found to be less susceptible than tgOv lines expressing ovine PrP at physiological levels. These data argue that ovine PrP(VRQ) provided a better substrate for sheep prion replication than did mouse PrP. Altogether, these tgOv mice could be an improved model for experimental studies on natural sheep scrapie. PMID- 11390600 TI - Nonspecific down-regulation of CD8+ T-cell responses in mice expressing human papillomavirus type 16 E7 oncoprotein from the keratin-14 promoter. AB - The E7 oncoprotein of human papillomavirus 16 (HPV16) transforms basal and suprabasal cervical epithelial cells and is a tumor-specific antigen in cervical carcinoma, to which immunotherapeutic strategies aimed at cytotoxic T-lymphocyte (CTL) induction are currently directed. By quantifying major histocompatibility complex class I tetramer-binding T cells and CTL in mice expressing an HPV16 E7 transgene from the keratin-14 (K14) promoter in basal and suprabasal keratinocytes and in thymic cortical epithelium, we show that antigen responsiveness of both E7- and non-E7-specific CD8+ cells is down-regulated compared to non-E7 transgenic control mice. We show that the effect is specific for E7, and not another transgene, expressed from the K14 promoter. Down regulation did not involve deletion of CD8+ T cells of high affinity or high avidity, and T-cell receptor (TCR) Vbeta-chain usage and TCR receptor density were similar in antigen-responsive cells from E7 transgenic and non-E7 transgenic mice. These data indicate that E7 expressed chronically from the K14 promoter nonspecifically down-regulates CD8+ T-cell responses. The in vitro data correlated with the failure of immunized E7 transgenic mice to control the growth of an E7-expressing tumor challenge. We have previously shown that E7-directed CTL down-regulation correlates with E7 expression in peripheral but not thymic epithelium (T. Doan et al., J. Virol. 73:6166-6170, 1999). The findings have implications for the immunological consequences of E7-expressing tumor development and E7-directed immunization strategies. Generically, the findings illustrate a T-cell immunomodulatory function for a virally encoded human oncoprotein. PMID- 11390601 TI - N-linked glycosylation sites adjacent to and within the V1/V2 and the V3 loops of dualtropic human immunodeficiency virus type 1 isolate DH12 gp120 affect coreceptor usage and cellular tropism. AB - The envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1) is extensively glycosylated, containing approximately 23 asparagine (N)-linked glycosylation sites on its gp120 subunit. In this study, specific glycosylation sites on gp120 of a dualtropic primary HIV-1 isolate, DH12, were eliminated by site-directed mutagenesis and the properties of the resulting mutant envelopes were evaluated using a recombinant vaccinia virus-based cell-to-cell fusion assay alone or in the context of viral infections. Of the glycosylation sites that were evaluated, those proximal to the V1/V2 loops (N135, N141, N156, N160) and the V3 loops (N301) of gp120 were functionally critical. The glycosylation site mutations near the V1/V2 loop compromised the use of CCR5 and CXCR4 equally. In contrast, a mutation within the V3 loop preferentially inhibited the usage of CCR5; although this mutant protein completely lost its CCR5-dependent fusion activity, it retained 50% of the wild-type fusion activity with CXCR4. The replication of a virus containing this mutation was severely compromised in peripheral blood mononuclear cells, MT-4 cells, and primary monocyte-derived macrophages. A revertant virus, which acquired second site changes in the V3 loop that resulted in an increase in net positive charge, was isolated. The revertant virus fully recovered the usage of CXCR4 but not of CCR5, thereby altering the tropism of the parental virus from dualtropic to T-tropic. These results suggest that carbohydrate moieties near the V1/V2 and the V3 loops play critical roles in maintaining proper conformation of the variable loops for optimal interaction with receptors. Our results, combined with those of previously reported studies, further demonstrate that the function of individual glycans may be virus isolate dependent. PMID- 11390602 TI - Mink cell focus-forming murine leukemia virus killing of mink cells involves apoptosis and superinfection. AB - Induction of apoptosis by different types of pathogenic retroviruses is an important step in disease development. We have observed that infection of thymic lymphocytes by the mink cell focus-forming murine leukemia virus (MCF MLV) during the preleukemic period resulted in an enhancement of apoptosis of these cells. To further study the ability of MCF MLVs to induce apoptosis and the role of this process in viral pathogenesis, we have developed an in vitro system of virus induced apoptosis. MCF13 MLV infection of mink epithelial cells resulted in the production of cytopathic foci. In contrast, infection of mink cells with the 4070A amphotropic MLV did not produce any cytopathic effects. Staining of MCF13 MLV-infected cells with propidium iodide and annexin V-fluorescein isothiocyanate indicated that virus-induced cell death was due to apoptosis. At 6 days postinfection, the percentage of apoptotic MCF13 MLV-infected cells was 27% compared with 2 to 3% for mock- or amphotropic MLV-infected cells, representing a 9- to 14-fold difference. Assays for caspase-3 activation confirmed the detection by flow cytometry of apoptosis of MCF13 MLV-infected cells. Large amounts of unintegrated linear viral DNA were detectable by Southern blot analysis during the acute phase of infection, which indicated that MCF13 MLV is able to superinfect mink cells. Unintegrated viral DNA of only the linear form was detectable in thymic lymphocytes isolated from MCF13 MLV-inoculated mice during the preleukemic period. These results indicated that the ability of MCF13 MLV to induce apoptosis is correlated with its ability to superinfect cells and that this occurs as an early step in thymic lymphoma development. PMID- 11390603 TI - Proper processing of avian sarcoma/leukosis virus capsid proteins is required for infectivity. AB - The formation of the mature carboxyl terminus of CA in avian sarcoma/leukemia virus is the result of a sequence of cleavage events at three PR sites that lie between CA and NC in the Gag polyprotein. The initial cleavage forms the amino terminus of the NC protein and releases an immature CA, named CA1, with a spacer peptide at its carboxyl terminus. Cleavage of either 9 or 12 amino acids from the carboxyl terminus creates two mature CA species, named CA2 and CA3, that can be detected in avian sarcoma/leukemia virus (R. B. Pepinsky, I. A. Papayannopoulos, E. P. Chow, N. K. Krishna, R. C. Craven, and V. M. Vogt, J. Virol. 69:6430-6438, 1995). To study the importance of each of the three CA proteins, we introduced amino acid substitutions into each CA cleavage junction and studied their effects on CA processing as well as virus assembly and infectivity. Preventing cleavage at any of the three sites produced noninfectious virus. In contrast, a mutant in which cleavage at site 1 was enhanced so that particles contained CA2 and CA3 but little detectable CA1 was infectious. These results support the idea that infectivity of the virus is closely linked to proper processing of the carboxyl terminus to form two mature CA proteins. PMID- 11390605 TI - Epstein-Barr virus SM protein interacts with mRNA in vivo and mediates a gene specific increase in cytoplasmic mRNA. AB - SM is an Epstein-Barr virus (EBV) gene expressed during early lytic replication of EBV. SM encodes a nuclear phosphoprotein that functions as a posttranscriptional regulator of gene expression. SM has been implicated in several aspects of gene regulation, including nuclear mRNA stabilization, posttranscriptional processing, and nuclear mRNA export. Activation by SM is promoter independent but gene specific. The mechanism by which SM selectively activates some EBV target genes or heterologous reporter genes remains to be determined. SM binds RNA in vitro, suggesting that sequence- or structure specific mRNA interactions might mediate SM specificity. We have further analyzed RNA binding by SM and demonstrated that proteolytic cleavage of SM and consequent exposure of an arginine-rich region are necessary to allow RNA binding in vitro. However, SM mutants with deletions of this arginine-rich region localized normally in the nucleus and were fully functional in gene activation. We therefore developed an assay to study in vivo interactions of SM with target mRNAs based on immunoprecipitation of SM from cell lysates followed by RNase protection analysis. Using this assay, we demonstrated that SM forms complexes with specific mRNAs in vivo. SM binds mRNAs from both SM-responsive as well as nonresponsive intronless genes and increases the nuclear accumulation of both types of mRNAs. In addition, SM preferentially associates with newly transcribed mRNAs. These data indicate that SM forms complexes with mRNAs in the nucleus and enhances their nuclear accumulation. However, SM does not enhance cytoplasmic accumulation of all transcripts that it binds to the same degree, suggesting that additional mRNA-specific characteristics, such as nuclear retention motifs or binding sites for cellular proteins, also determine responsiveness to SM. PMID- 11390604 TI - Human cytomegalovirus up-regulates the phosphatidylinositol 3-kinase (PI3-K) pathway: inhibition of PI3-K activity inhibits viral replication and virus induced signaling. AB - Infection of quiescent fibroblasts with human cytomegalovirus (HCMV) was found to cause a rapid activation of cellular phosphatidylinositol 3-kinase (PI3-K). Maximum PI3-K activation occurred from 15 to 30 min postinfection. This activation was transient, and by 2 h postinfection (hpi), PI3-K activity had declined to preinfection levels. However, at 4 hpi, a second tier of PI3-K activation was detected, and PI3-K activity remained elevated relative to that of mock-infected cells for the remainder of infection. The cellular kinases Akt and p70S6K and the transcription factor NF-kappaB were activated in a PI3-K-dependent manner at similar times following HCMV infection. Analysis using UV-irradiated virus indicated that no viral protein synthesis was necessary for the first phase of PI3-K activation, but viral protein expression was required for the second tier of PI3-K activation. Treatment of infected fibroblasts with LY294002, a potent and specific inhibitor of PI3-K kinase activity, caused a 4-log decrease in viral titers. LY294002 did not inhibit viral entry, but it did decrease viral immediate-early gene expression. In addition, the protein levels of two viral early genes required for DNA replication, UL84 and UL44, were significantly lower in the presence of LY294002. Furthermore, viral DNA replication was strongly inhibited by LY294002 treatment. This inhibition of viral DNA replication could be reversed by adding back the products of PI3-K activity (PI-3,4-P(2) and PI 3,4,5-P(3)), demonstrating that the effect of LY294002 on the viral life cycle was specifically due to the inhibition of PI3-K activity. These results are the first to suggest that PI3-K mediates HCMV-induced activation of host cell mitogenic pathways. They also provide strong evidence that PI3-K activation is important for initiation of viral DNA replication and completion of the viral lytic life cycle. PMID- 11390606 TI - Oligomerization mediated by a helix-loop-helix-like domain of baculovirus IE1 is required for early promoter transactivation. AB - IE1 is a principal transcriptional regulator of Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). Transactivation by IE1 is stimulated when early viral promoters are cis linked to homologous-region (hr) enhancer sequences of AcMNPV. This transcriptional enhancement is correlated with the binding of IE1 as a dimer to the 28-bp palindromic repeats comprising the hr enhancer. To define the role of homophilic interactions in IE1 transactivation, we have mapped the IE1 domains required for oligomerization. We report here that IE1 oligomerizes by a mechanism independent of enhancer binding, as demonstrated by in vitro pull-down assays using fusions of IE1 (582 residues) to the C terminus of glutathione S-transferase. In vivo oligomerization of IE1 was verified by immunoprecipitation of IE1 complexes from extracts of plasmid transfected SF21 cells. Analyses of a series of site-directed IE1 insertion mutations indicated that a helix-loop-helix (HLH)-like domain extending from residue 543 to residue 568 is the primary determinant of oligomerization. Replacement of residues within the hydrophobic face of the putative dimerization domain disrupted IE1 homophilic interactions and caused loss of IE1 transactivation of hr-dependent promoters in plasmid transfection assays. Thus, oligomerization is required for IE1 transcriptional stimulation. HLH mutations also reduced IE1 stability and abrogated transactivation of non-hr-dependent promoters. These data support a model wherein IE1 oligomerizes prior to DNA binding to facilitate proper interaction with the symmetrical recognition sites within the hr enhancer and thereby promote the transcription of early viral genes. PMID- 11390607 TI - Trypsin cleavage stabilizes the rotavirus VP4 spike. AB - Trypsin enhances rotavirus infectivity by an unknown mechanism. To examine the structural basis of trypsin-enhanced infectivity in rotaviruses, SA11 4F triple layered particles (TLPs) grown in the absence (nontrypsinized rotavirus [NTR]) or presence (trypsinized rotavirus [TR]) of trypsin were characterized to determine the structure, the protein composition, and the infectivity of the particles before and after trypsin treatment. As expected, VP4 was not cleaved in NTR particles and was cleaved into VP5(*) and VP8(*) in TR particles. However, surprisingly, while the VP4 spikes were clearly visible and well ordered in the electron cryomicroscopy reconstructions of TR TLPs, they were totally absent in the reconstructions of NTR TLPs. Biochemical analysis with radiolabeled particles indicated that the stoichiometry of the VP4 in NTR particles was the same as that in TR particles and that the VP8(*) portion of NTR, but not TR, particles is susceptible to further proteolysis by trypsin. Taken together, these structural and biochemical data show that the VP4 spikes in the NTR TLPs are icosahedrally disordered and that they are conformationally different. Structural studies on the NTR TLPs after trypsin treatment showed that spike structure could be partially recovered. Following additional trypsin treatment, infectivity was enhanced for both NTR and TR particles, but the infectivity of NTR remained 2 logs lower than that of TR particles. Increased infectivity in these particles corresponded to additional cleavages in VP5(*), at amino acids 259, 583, and putatively 467, which are conserved in all P serotypes of human and animal group A rotaviruses and also corresponded with a structural change in VP7. These biochemical and structural results show that trypsin cleavage imparts order to VP4 spikes on de novo synthesized virus particles, and these ordered spikes make virus entry into cells more efficient. PMID- 11390608 TI - Identification of a novel transcriptional repressor encoded by human cytomegalovirus. AB - The expression of human cytomegalovirus (HCMV) genes during viral replication is precisely regulated, with the interactions of both transcriptional activators and repressors determining the level of gene expression. One gene of HCMV, the US3 gene, is transcriptionally repressed early in infection. Repression of US3 expression requires viral infection and protein synthesis and is mediated through a DNA sequence, the transcriptional repressive element. In this report, we identify the protein that represses US3 transcription as the product of the HCMV UL34 open reading frame. The protein encoded by UL34 (pUL34) binds to the US3 transcriptional repressive element in yeast and in vitro. pUL34 localizes to the nucleus and alone is sufficient for repression of US3 expression. The data presented here, along with earlier data (B. J. Biegalke, J. Virol. 72:5457-5463, 1998), suggests that pUL34 binding of the transcriptional repressive element prevents transcription initiation complex formation. PMID- 11390609 TI - Hantavirus infection induces the expression of RANTES and IP-10 without causing increased permeability in human lung microvascular endothelial cells. AB - Sin Nombre virus (SNV) and Hantaan virus (HTN) infect endothelial cells and are associated with different patterns of increased vascular permeability during human disease. It is thought that such patterns of increased vascular permeability are a consequence of endothelial activation and subsequent dysfunction mediated by differential immune responses to hantavirus infection. In this study, the ability of hantavirus to directly induce activation of human lung microvascular endothelial cells (HMVEC-Ls) was examined. No virus-specific modulation in the constitutive or cytokine-induced expression of cellular adhesion molecules (CD40, CD54, CD61, CD62E, CD62P, CD106, and major histocompatibility complex classes I and II) or in cytokines and chemokines (eotaxin, tumor necrosis factor alpha, interleukin 1beta [IL-1beta], IL-6, IL-8, MCP-1, MIP-1alpha, and MIP-1beta) was detected at either the protein or message level in hantavirus-infected HMVEC-Ls. Furthermore, no virus-specific enhancement of paracellular or transcellular permeability or changes in the organization and distribution of endothelial intercellular junctional proteins was observed. However, infection with either HTN or SNV resulted in detectable levels of the chemokines RANTES and IP-10 (the 10-kDa interferon-inducible protein) in HMVEC-Ls within 72 h and was associated with nuclear translocation of interferon regulatory factor 3 (IRF-3) and IRF-7. Gamma interferon (IFN-gamma)-induced expression of RANTES and IP-10 could also be detected in uninfected HMVEC-Ls and was associated with nuclear translocation of IRF-1 and IRF-3. Treatment of hantavirus-infected HMVEC-Ls with IFN-gamma for 24 h resulted in a synergistic enhancement in the expression of both RANTES and IP-10 and was associated with nuclear translocation of IRF-1, IRF-3, IRF-7, and NF-kappaB p65. These results reveal a possible mechanism by which hantavirus infection and a TH1 immune response can cooperate to synergistically enhance chemokine expression by HMVEC Ls and trigger immune-mediated increases in vascular permeability. PMID- 11390610 TI - Free major histocompatibility complex class I heavy chain is preferentially targeted for degradation by human T-cell leukemia/lymphotropic virus type 1 p12(I) protein. AB - Human T-cell leukemia virus type 1 (HTLV-1) establishes a persistent infection in the host despite a vigorous virus-specific immune response. Here we demonstrate that an HTLV-1-encoded protein, p12(I), resides in the endoplasmic reticulum (ER) and Golgi and physically binds to the free human major histocompatibility complex class I heavy chains (MHC-I-Hc) encoded by the HLA-A2, -B7, and -Cw4 alleles. As a result of this interaction, the newly synthesized MHC-I-Hc fails to associate with beta(2)-microglobulin and is retrotranslocated to the cytosol, where it is degraded by the proteasome complex. Targeting of the free MHC-I-Hc, and not the MHC-I-Hc-beta(2)-microglobulin complex, by p12(I) represents a novel mechanism of viral interference and disrupts the intracellular trafficking of MHC-I, which results in a significant decrease in surface levels of MHC-I on human T-cells. These findings suggest that the interaction of p12(I) with MHC-1-Hc may interfere with antigen presentation in vivo and facilitate escape of HTLV-1-infected cells from immune recognition. PMID- 11390612 TI - Rearrangement of the genes of vesicular stomatitis virus eliminates clinical disease in the natural host: new strategy for vaccine development. AB - Gene expression among the nonsegmented negative-strand RNA viruses is controlled by distance from the single transcriptional promoter, so the phenotypes of these viruses can be systematically manipulated by gene rearrangement. We examined the potential of gene rearrangement as a means to develop live attenuated vaccine candidates against Vesicular stomatitis virus (VSV) in domestic swine, a natural host for this virus. The results showed that moving the nucleocapsid protein gene away from the single transcriptional promoter attenuated and ultimately eliminated the potential of the virus to cause disease. Combining this change with relocation of the surface glycoprotein gene yielded a vaccine that protected against challenge with wild-type VSV. By incremental manipulation of viral properties, gene rearrangement provides a new approach to generating live attenuated vaccines against this class of virus. PMID- 11390611 TI - Hepatitis C virus nonstructural 5A protein induces interleukin-8, leading to partial inhibition of the interferon-induced antiviral response. AB - Hepatitis C virus (HCV), a major cause of liver disease worldwide, is frequently resistant to the antiviral alpha interferon (IFN). The HCV nonstructural 5A (NS5A) protein has been implicated in HCV antiviral resistance in many studies. NS5A antagonizes the IFN antiviral response in vitro, and one mechanism is via inhibition of a key IFN-induced enzyme, the double-stranded-RNA-activated protein kinase (PKR). In the present study we determined if NS5A uses other strategies to subvert the IFN system. Expression of full-length NS5A proteins from patients who exhibited a complete response (FL-NS5A-CR) or were nonresponsive (FL-NS5A-NR) to IFN therapy in HeLa cells had no effect on IFN induction of IFN-stimulated gene factor 3 (ISGF-3). Expression of mutant NS5A proteins lacking 110 (NS5A DeltaN110), 222 (NS5A-DeltaN222), and 334 amino-terminal amino acids and mutants lacking 117 and 230 carboxy-terminal amino acids also had no effect on ISGF-3 induction by IFN. Expression of FL-NS5A-CR and FL-NS5A-NR did not affect IFN induced STAT-1 tyrosine phosphorylation or upregulation of PKR and major histocompatibility complex class I antigens. However, NS5A expression in human cells induced interleukin 8 (IL-8) mRNA and protein, and this effect correlated with inhibition of the antiviral effects of IFN in an in vitro bioassay. NS5A induced transcription of a reporter gene driven by the IL-8 promoter, and the first 133 bp of the IL-8 promoter made up the minimal domain required for NS5A transactivation. NS5A-DeltaN110 and NS5A-DeltaN222 stimulated the IL-8 promoter to higher levels than did the full-length NS5A protein, and this correlated with increased nuclear localization of the proteins. Additional mutagenesis of the IL 8 promoter suggested that NF-kappaB and AP-1 were important in NS5A-DeltaN222 transactivation in the presence of tumor necrosis factor alpha and that NF-IL-6 was inhibitory to this process. This study suggests that NS5A inhibits the antiviral actions of IFN by at least two mechanisms and provides the first evidence for a biological effect of the transcriptional activity of the NS5A protein. During HCV infection, viral proteins may induce chemokines that contribute to HCV antiviral resistance and pathogenesis. PMID- 11390613 TI - Axonal damage is T cell mediated and occurs concomitantly with demyelination in mice infected with a neurotropic coronavirus. AB - Mice infected with mouse hepatitis virus (MHV) strain JHM develop primary demyelination. Herein we show that axonal damage occurred in areas of demyelination and also in adjacent areas devoid of myelin damage. Immunodeficient MHV-infected RAG1-/- mice (mice defective in recombinase activating gene 1 expression) do not develop demyelination unless they receive splenocytes from a mouse previously immunized against MHV (G. F. Wu, A. Dandekar, L. Pewe, and S. Perlman, J. Immunol. 165:2278-2286, 2000). In the present study, we show that adoptive transfer of T cells was also required for the majority of the axonal injury observed in these animals. Both demyelination and axonal damage were apparent by 7 days posttransfer. Recent data suggest that axonal injury is a major factor in the long-term disability observed in patients with multiple sclerosis. Our data demonstrate that immune system-mediated damage to axons is also a common feature in mice with MHV-induced demyelination. Remarkably, there appeared to be a minimal, if any, interval of time between the appearance of demyelination and that of axonal injury. PMID- 11390614 TI - p21cip1 Degradation in differentiated keratinocytes is abrogated by costabilization with cyclin E induced by human papillomavirus E7. AB - The human papillomavirus (HPV) E7 protein promotes S-phase reentry in a fraction of postmitotic, differentiated keratinocytes. Here we report that these cells contain an inherent mechanism that opposes E7-induced DNA replication. In organotypic raft cultures of primary human keratinocytes, neither cyclin E nor p21cip1 is detectable in situ. However, E7-transduced differentiated cells not in S phase accumulate abundant cyclin E and p21cip1. We show that normally p21cip1 protein is rapidly degraded by proteasomes. In the presence of E7 or E6/E7, p21cip1, cyclin E, and cyclin E2 proteins were all up-regulated. The accumulation of p21cip1 protein is a posttranscriptional event, and ectopic cyclin E expression was sufficient to trigger it. In constract, cdk2 and p27kip1 were abundant in normal differentiated cells and were not significantly affected by E7. Cyclin E, cdk2, and p21cip1 or p27kip1 formed complexes, and relatively little kinase activity was found associated with cyclin E or cdk2. In patient papillomas and E7 raft cultures, all p27kip1-positive cells were negative for bromodeoxyuridine (BrdU) incorporation, but only some also contained cyclin E and p21cip1. In contrast, all cyclin E-positive cells also contained p27kip1. When the expression of p21cip1 was reduced by rottlerin, a PKC delta inhibitor, p27kip1- and BrdU-positive cells remained unchanged. These observations show that high levels of endogenous p27kip1 can prevent E7-induced S-phase reentry. This inhibition then leads to the stabilization of cyclin E and p21cip1. Since efficient initiation of viral DNA replication requires cyclin E and cdk2, its inhibition accounts for heterogeneous viral activities in productively infected lesions. PMID- 11390615 TI - Epstein-Barr virus immediate-early protein BRLF1 induces the lytic form of viral replication through a mechanism involving phosphatidylinositol-3 kinase activation. AB - Expression of the Epstein-Barr virus (EBV) immediate-early (IE) protein BRLF1 induces the lytic form of viral replication in most EBV-positive cell lines. BRLF1 is a transcriptional activator that binds directly to a GC-rich motif present in some EBV lytic gene promoters. However, BRLF1 activates transcription of the other IE protein, BZLF1, through an indirect mechanism which we previously showed to require activation of the stress mitogen-activated protein kinases. Here we demonstrate that BRLF1 activates phosphatidylinositol-3 (PI3) kinase signaling in host cells. We show that the specific PI3 kinase inhibitor, LY294002, completely abrogates the ability of a BRLF1 adenovirus vector to induce the lytic form of EBV infection, while not affecting lytic infection induced by a BZLF1 adenovirus vector. Furthermore, we demonstrate that the requirement for PI3 kinase activation in BRLF1-induced transcriptional activation is promoter dependent. BRLF1 activation of the SM early promoter (which occurs through a direct binding mechanism) does not require PI3 kinase activation, whereas activation of the IE BZLF1 and early BMRF1 promoters requires PI3 kinase activation. Thus, there are clearly two separate mechanisms by which BRLF1 induces transcriptional activation. PMID- 11390616 TI - ICP0, ICP4, or VP16 expressed from adenovirus vectors induces reactivation of latent herpes simplex virus type 1 in primary cultures of latently infected trigeminal ganglion cells. AB - In a previous study, we demonstrated that infected-cell polypeptide 0 (ICP0) is necessary for the efficient reactivation of herpes simplex virus type 1 (HSV-1) in primary cultures of latently infected trigeminal ganglion (TG) cells (W. P. Halford and P. A. Schaffer, J. Virol. 75:3240-3249, 2001). The present study was undertaken to determine whether ICP0 is sufficient to trigger HSV-1 reactivation in latently infected TG cells. To test this hypothesis, replication-defective adenovirus vectors that express wild-type and mutant forms of ICP0 under the control of a tetracycline response element (TRE) promoter were constructed. Similar adenovirus vectors encoding wild-type ICP4, wild-type and mutant forms of the HSV-1 origin-binding protein (OBP), and wild-type and mutant forms of VP16 were also constructed. The TRE promoter was induced by coinfection of Vero cells with the test vector and an adenovirus vector that expresses the reverse tetracycline-regulated transactivator in the presence of doxycycline. Northern blot analysis demonstrated that transcription of the OBP gene in the adenovirus expression vector increased as a function of doxycycline concentration over a range of 0.1 to 10 microM. Likewise, Western blot analysis demonstrated that addition of 3 microM doxycycline to adenovirus vector-infected Vero cells resulted in a 100-fold increase in OBP expression. Wild-type forms of ICP0, ICP4, OBP, and VP16 expressed from adenovirus vectors were functional based on their ability to complement plaque formation in Vero cells by replication-defective HSV 1 strains with mutations in these genes. Adenovirus vectors that express wild type forms of ICP0, ICP4, or VP16 induced reactivation of HSV-1 in 86% +/- 5%, 86% +/- 5%, and 97% +/- 5% of TG cell cultures, respectively (means +/- standard deviations). In contrast, vectors that express wild-type OBP or mutant forms of ICP0, OBP, or VP16 induced reactivation in 5% +/- 5%, 8% +/- 0%, 0% +/- 0%, and 13% +/- 6% of TG cell cultures, respectively. In control infections, an adenovirus vector expressed green fluorescent protein efficiently in TG neurons but did not induce HSV-1 reactivation. Therefore, expression of ICP0, ICP4, or VP16 is sufficient to induce HSV-1 reactivation in latently infected TG cell cultures. We conclude that this system provides a powerful tool for determining which cellular and viral proteins are sufficient to induce HSV-1 reactivation from neuronal latency. PMID- 11390617 TI - Formation of wild-type and chimeric influenza virus-like particles following simultaneous expression of only four structural proteins. AB - We are studying the structural proteins and molecular interactions required for formation and release of influenza virus-like particles (VLPs) from the cell surface. To investigate these events, we generated a quadruple baculovirus recombinant that simultaneously expresses in Sf9 cells the hemagglutinin (HA), neuraminidase (NA), matrix (M1), and M2 proteins of influenza virus A/Udorn/72 (H3N2). Using this quadruple recombinant, we have been able to demonstrate by double-labeling immunofluorescence that matrix protein (M1) localizes in nuclei as well as at discrete areas of the plasma membrane where HA and NA colocalize at the cell surface. Western blot analysis of cell supernatant showed that M1, HA, and NA were secreted into the culture medium. Furthermore, these proteins comigrated in similar fractions when concentrated supernatant was subjected to differential centrifugation. Electron microscopic examination (EM) of these fractions revealed influenza VLPs bearing surface projections that closely resemble those of wild-type influenza virus. Immunogold labeling and EM demonstrated that the HA and NA were present on the surface of the VLPs. We further investigated the minimal number of structural proteins necessary for VLP assembly and release using single-gene baculovirus recombinants. Expression of M1 protein alone led to the release of vesicular particles, which in gradient centrifugation analysis migrated in a similar pattern to that of the VLPs. Immunoprecipitation of M1 protein from purified M1 vesicles, VLPs, or influenza virus showed that the relative amount of M1 protein associated with M1 vesicles or VLPs was higher than that associated with virions, suggesting that particle formation and budding is a very frequent event. Finally, the HA gene within the quadruple recombinant was replaced either by a gene encoding the G protein of vesicular stomatitis virus or by a hybrid gene containing the cytoplasmic tail and transmembrane domain of the HA and the ectodomain of the G protein. Each of these constructs was able to drive the assembly and release of VLPs, although enhanced recruitment of the G glycoprotein onto the surface of the particle was observed with the recombinant carrying a G/HA chimeric gene. The described approach to assembly of wild-type and chimeric influenza VLPs may provide a valuable tool for further investigation of viral morphogenesis and genome packaging as well as for the development of novel vaccines. PMID- 11390618 TI - The domains of glycoprotein D required to block apoptosis depend on whether glycoprotein D is present in the virions carrying herpes simplex virus 1 genome lacking the gene encoding the glycoprotein. AB - An earlier report showed that viruses lacking the open reading frames encoding glycoproteins J and D but containing the glycoprotein D in their envelopes (gD-/+ stocks) and viruses lacking both the open reading frames and the glycoproteins in their envelopes (gD-/- stocks) induce apoptosis (G. Zhou, V. Galvan, G. Campadelli-Fiume, and B. Roizman, J. Virol. 74:11782-11791, 2000). Furthermore, apoptosis was blocked by delivery in trans of genes expressing glycoprotein D or J. Whereas gD-/- stocks attach but cannot initiate productive infection, gD-/+ stocks infect cells and produce gD-/- progeny virus. The difference in the infectivity of these two stocks suggested the possibility that the requirements for blocking apoptosis may be different. To test this hypothesis, we cloned into baculoviruses the entire wild-type glycoprotein D (Bac-gD-WT), the ectodomain only (Bac-gD-A), the ectodomain and the transmembrane domain (Bac-gD-B), the ectodomain and the cytoplasmic domain without the transmembrane domain (Bac-gD C), or the transmembrane domain and the carboxyl-terminal cytoplasmic domain (Bac gD-D). We report the following. Apoptosis induced by gD-/+ stocks was blocked by delivery in trans of recombinant baculovirus Bac-gD-WT, Bac-gD-A, Bac-gD-B, or Bac-gD-C but not of Bac-gD. Apoptosis induced by gD-/- stocks was blocked by Bac gD-WT or by a mixture of Bac-gD-B and Bac-gD-D but not by any baculoviruses expressing truncated glycoprotein D alone or by the mixture of Bac-gD-A and Bac gD-D. We conclude that the requirements to block apoptosis induced by the two virus stocks are different. The gD ectodomain is sufficient to block apoptosis induced by gD, whereas both the ectodomain and the cytoplasmic domain are required to block apoptosis induced by gD-/- stocks. The results indicate that in the case of gD-/- stocks, the transmembrane domain is required either to deliver the ectodomain to the appropriate intracellular compartment or to form multimeric constructs which virtually reconstitute gD through the interaction of transmembrane domains. PMID- 11390619 TI - Differential incorporation of CD45, CD80 (B7-1), CD86 (B7-2), and major histocompatibility complex class I and II molecules into human immunodeficiency virus type 1 virions and microvesicles: implications for viral pathogenesis and immune regulation. AB - Human immunodeficiency virus (HIV) infection results in a functional impairment of CD4(+) T cells long before a quantitative decline in circulating CD4(+) T cells is evident. The mechanism(s) responsible for this functional unresponsiveness and eventual depletion of CD4(+) T cells remains unclear. Both direct effects of cytopathic infection of CD4(+) cells and indirect effects in which uninfected "bystander" cells are functionally compromised or killed have been implicated as contributing to the immunopathogenesis of HIV infection. Because T-cell receptor engagement of major histocompatibility complex (MHC) molecules in the absence of costimulation mediated via CD28 binding to CD80 (B7 1) or CD86 (B7-2) can lead to anergy or apoptosis, we determined whether HIV type 1 (HIV-1) virions incorporated MHC class I (MHC-I), MHC-II, CD80, or CD86. Microvesicles produced from matched uninfected cells were also evaluated. HIV infection increased MHC-II expression on T- and B-cell lines, macrophages, and peripheral blood mononclear cells (PBMC) but did not significantly alter the expression of CD80 or CD86. HIV virions derived from all MHC-II-positive cell types incorporated high levels of MHC-II, and both virions and microvesicles preferentially incorporated CD86 compared to CD80. CD45, expressed at high levels on cells, was identified as a protein present at high levels on microvesicles but was not detected on HIV-1 virions. Virion-associated, host cell-derived molecules impacted the ability of noninfectious HIV virions to trigger death in freshly isolated PBMC. These results demonstrate the preferential incorporation or exclusion of host cell proteins by budding HIV-1 virions and suggest that host cell proteins present on HIV-1 virions may contribute to the overall pathogenesis of HIV-1 infection. PMID- 11390620 TI - Varicella-zoster virus infection of human dendritic cells and transmission to T cells: implications for virus dissemination in the host. AB - During primary varicella-zoster virus (VZV) infection, it is presumed that virus is transmitted from mucosal sites to regional lymph nodes, where T cells become infected. The cell type responsible for VZV transport from the mucosa to the lymph nodes has not been defined. In this study, we assessed the susceptibility of human monocyte-derived dendritic cells to infection with VZV. Dendritic cells were inoculated with the VZV strain Schenke and assessed by flow cytometry for VZV and dendritic cell (CD1a) antigen expression. In five replicate experiments, 34.4% +/- 6.6% (mean +/- SEM) of CD1a(+) cells were also VZV antigen positive. Dendritic cells were also shown to be susceptible to VZV infection by the detection of immediate-early (IE62), early (ORF29), and late (gC) gene products in CD1a(+) dendritic cells. Infectious virus was recovered from infected dendritic cells, and cell-to-cell contact was required for transmission of virus to permissive fibroblasts. VZV-infected dendritic cells showed no significant decrease in cell viability or evidence of apoptosis and did not exhibit altered cell surface levels of major histocompatibility complex (MHC) class I, MHC class II, CD86, CD40, or CD1a. Significantly, when autologous T lymphocytes were incubated with VZV-infected dendritic cells, VZV antigens were readily detected in CD3(+) T lymphocytes and infectious virus was recovered from these cells. These data provide the first evidence that dendritic cells are permissive to VZV and that dendritic cell infection can lead to transmission of virus to T lymphocytes. These findings have implications for our understanding of how virus may be disseminated during primary VZV infection. PMID- 11390621 TI - Viral interferon regulatory factor 1 of Kaposi's sarcoma-associated herpesvirus binds to p53 and represses p53-dependent transcription and apoptosis. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) is related to the development of Kaposi's sarcoma. Open reading frame K9 of KSHV encodes viral interferon regulatory factor 1 (vIRF1), which functions as a repressor of interferon- and IRF1-mediated signal transduction. In addition, vIRF1 acts as an oncogene to induce cellular transformation. Here we show that vIRF1 directly associates with the tumor suppressor p53 and represses its functions. The vIRF1 interaction domains of p53 are the DNA binding domain (amino acids [aa] 100 to 300) and the tetramerization domain (aa 300 to 393). p53 interacts with the central region (aa 152 to 360) of vIRF1. vIRF1 suppresses p53-dependent transcription and deregulates its apoptotic activity. These results suggest that vIRF1 may regulate cellular function by inhibiting p53. PMID- 11390622 TI - Hybrid vectors based on adeno-associated virus serotypes 2 and 5 for muscle directed gene transfer. AB - Vectors based on hybrids consisting of adeno-associated virus types 2 (ITRs and Rep) and 5 (Cap) were evaluated for muscle-directed gene transfer (called AAV2/5). Evaluation in immune-competent mice revealed greater transduction efficacy with AAV2/5 than with AAV2 and no cross-neutralization between AAV2/5 and AAV2. Interestingly, we saw no immunologic evidence of previous exposure to AAV5 capsids in a large population of healthy human subjects. PMID- 11390623 TI - Role of the promyelocytic leukemia protein PML in the interferon sensitivity of lymphocytic choriomeningitis virus. AB - Lymphocytic choriomeningitis virus (LCMV) induces type I interferon (alpha and beta interferon [IFN-alpha and IFN-beta]) upon infection and yet is sensitive to the addition of type II interferon (gamma interferon [IFN-gamma]) to the culture media. This sensitivity is biologically important because it correlates inversely with the ability of certain LCMV strains to persist in mice (D. Moskophidis, M. Battegay, M. A. Bruendler, E. Laine, I. Gresser, and R. M. Zinkernagel, J. Virol. 68:1951-1955, 1994). The cellular oncoprotein PML is induced by both IFN alpha/beta and IFN-gamma, and PML binds the LCMV Z protein and becomes redistributed within cells from nucleus to cytoplasm upon LCMV infection. In the present study, increased PML expression results in diminished LCMV replication, implicating PML in the IFN sensitivity of LCMV. Virus production in PML -/- murine embryonic fibroblasts (MEF) exceeds virus production in PML +/+ MEF, and this difference is exacerbated by IFN treatment that upregulates PML expression. IFN-gamma also diminishes LCMV production in PML -/- cells; therefore, viral IFN sensitivity is not entirely due to PML. Both viral mRNA production and viral protein production decrease as PML expression increases. Here we propose that PML reduces LCMV transcription through its interaction with the Z protein. PMID- 11390624 TI - Elevated levels of interleukin-8 in serum are associated with hepatitis C virus infection and resistance to interferon therapy. AB - Hepatitis C virus (HCV), a major cause of liver disease worldwide, is frequently resistant to the antiviral alpha interferon (IFN). We have recently found that the HCV NS5A protein induces expression of the proinflammatory chemokine IL-8 to partially inhibit the antiviral actions of IFN in vitro. To extend these observations, in the present study we examined the relationship between levels of IL-8 in serum, HCV infection, and biochemical response to IFN therapy. Levels of IL-8 were significantly elevated in 132 HCV-infected patients compared to levels in 32 normal healthy subjects and were also significantly higher in patients who did not respond to IFN therapy than in patients who did respond to therapy. This study suggests that HCV-induced changes in levels of chemokine and cytokine expression may be involved in HCV antiviral resistance, persistence, and pathogenesis. PMID- 11390625 TI - Expression of Moloney murine leukemia virus RNase H rescues the growth defect of an Escherichia coli mutant. AB - A 157-amino-acid fragment of Moloney murine leukemia virus reverse transcriptase encoding RNase H is shown to rescue the growth-defective phenotype of an Escherichia coli mutant. In vitro assays of the recombinant wild-type protein purified from the conditionally defective mutant confirm that it is catalytically active. Mutagenesis of one of the presumptive RNase H-catalytic residues results in production of a protein variant incapable of rescue and which lacks activity in vitro. Analyses of additional active site mutants demonstrate that their encoded variant proteins lack robust activity yet are able to rescue the bacterial mutant. These results suggest that genetic complementation may be useful for in vivo screening of mutant viral RNase H gene fragments and in evaluating their function under conditions that more closely mimic physiological conditions. The rescue system may also be useful in verifying the functional outcomes of mutations based on protein structural predictions and modeling. PMID- 11390626 TI - Heparin inhibits retrovirus binding to fibronectin as well as retrovirus gene transfer on fibronectin fragments. AB - Fibronectin fragments have been shown to improve retrovirus gene transfer efficiency by binding retrovirus and target cells. Using a novel virus adhesion assay, we confirmed binding of type C oncoretrovirus vectors to the heparin II domain of fibronectin and demonstrated inhibition of viral binding and gene transfer by heparin. PMID- 11390627 TI - Novel baculovirus expression vectors that provide sialylation of recombinant glycoproteins in lepidopteran insect cells. AB - This report describes novel baculovirus vectors designed to express mammalian beta1,4-galactosyltransferase and alpha2,6-sialyltransferase genes at early times after infection. Sf9 cells infected with these viral vectors, unlike cells infected with a wild-type baculovirus, produced a sialylated viral glycoprotein during the late phase of infection. Thus, the two mammalian glycosyltransferases encoded by these viral vectors are necessary and sufficient for sialylation of a foreign glycoprotein in insect cells under the conditions used in this study. While some of the new baculovirus vectors described in this study produced less, one produced wild-type levels of infectious budded virus progeny. PMID- 11390629 TI - Variant chromatin structure of the oriP region of Epstein-Barr virus and regulation of EBER1 expression by upstream sequences and oriP. AB - Most of the Epstein-Barr virus genome in latently infected cells is in a standard nucleosomal structure, but the region encompassing oriP and the Epstein-Barr virus-encoded small RNA (EBER) genes shows a distinctive pattern when digested with micrococcal nuclease. This pattern corresponds to a previously mapped nuclear matrix attachment region. Although the EBER genes are adjacent to oriP, there is only a two- to fourfold effect of oriP on EBER expression. However, sequences containing a consensus ATF site upstream of EBER1 are important for EBER1 expression. PMID- 11390628 TI - Epstein-Barr virus immediate-early protein BRLF1 interacts with CBP, promoting enhanced BRLF1 transactivation. AB - The Epstein-Barr virus (EBV) immediate-early protein BRLF1 is a transcriptional activator that mediates the switch from latent to lytic viral replication. Many transcriptional activators function, in part, due to an interaction with histone acetylases, such as CREB-binding protein (CBP). Here we demonstrate that BRLF1 interacts with the amino and carboxy termini of CBP and that multiple domains of the BRLF1 protein are necessary for this interaction. Furthermore, we show that the interaction between BRLF1 and CBP is important for BRLF1-induced activation of the early lytic EBV gene SM in Raji cells. PMID- 11390630 TI - Interleukin-1 receptor antagonist gene polymorphism and circulating levels of human immunodeficiency virus type 1 RNA in Brazilian women. AB - Interleukin-1 receptor antagonist (IL-1ra) gene polymorphisms in 83 human immunodeficiency virus (HIV)-seropositive women were evaluated. Fourteen of the subjects (16.9%) were homozygous for IL-1ra allele 2 (IL-1RN*2). These women had a lower median level of HIV RNA than did women homozygous for allele 1 (IL-1RN*1) (P = 0.01) or heterozygous for both alleles (P = 0.04). Among 46 subjects not receiving antiretroviral treatment, HIV levels were also reduced in IL-1RN*2 homozygous individuals (P < 0.05). There was no relation between IL-1ra alleles and CD4 levels. PMID- 11390631 TI - Kaposi's sarcoma-associated herpesvirus open reading frame 50 represses p53 induced transcriptional activity and apoptosis. AB - Kaposi's sarcoma-associated herpesvirus (KSHV) open reading frame 50 (ORF50) encodes a viral transcriptional activator which stimulates the transcription of viral early and late genes of KSHV. Here we show that ORF50 represses transcriptional activity of p53 and p53-induced apoptosis through interaction with CREB binding protein (CBP). This inhibitory effect of ORF50 on the transcriptional activity of p53 was relieved by the addition of CBP. ORF50 mutants, which are defective in interaction with CBP, lost the inhibitory effects on p53. Our data provide a framework for delineating the regulatory mechanisms used by KSHV to modulate cellular transcription and the cell cycle. PMID- 11390632 TI - Cellular and molecular mechanisms of estrogen regulation of memory function and neuroprotection against Alzheimer's disease: recent insights and remaining challenges. AB - This review focuses on recent advances in our knowledge of estrogen action in the brain. The greatest amount of attention was devoted to those studies that impact our understanding of estrogen regulation of memory function and prevention of degenerative diseases associated with memory systems, such as Alzheimer's disease. A review of recent advances in our understanding of estrogen receptors, both nuclear and membrane, is also presented. Finally, these data are considered in regard to their relevancy to the use of estrogen replacement therapy for cognitive health throughout menopause and the development of an estrogen replacement therapy designed for the unique requirements of the brain. PMID- 11390633 TI - Integrating orbitofrontal cortex into prefrontal theory: common processing themes across species and subdivisions. AB - Currently, many theories highlight either representational memory or rule representation as the hallmark of prefrontal function. Neurophysiological findings in the primate dorsolateral prefrontal cortex indicate that both features may characterize prefrontal processing. Neurons in the dorsolateral prefrontal cortex encode information in working memory, and this information is represented when relevant to the rules governing performance in a task. In this review, we discuss recent reports of encoding in primate and rat orbitofrontal regions indicating that these features also characterize activity in the orbitofrontal subdivision of the prefrontal cortex. These data indicate that (1) neural activity in the orbitofrontal cortex links the current incentive value of reinforcers to cues, rather than representing the physical features of cues or associated reinforcers; (2) this incentive-based information is represented in the orbitofrontal cortex when it is relevant to the rules guiding performance in a task; and (3) incentive information is also represented in the orbitofrontal cortex in working memory during delays when neither the cues nor reinforcers are present. Therefore, although the orbitofrontal cortex appears to be uniquely specialized to process incentive or motivational information, it may be integrated into a more global framework of prefrontal function characterized by representational encoding of performance-relevant information. PMID- 11390634 TI - Contextual and auditory fear conditioning are mediated by the lateral, basal, and central amygdaloid nuclei in rats. AB - A large body of literature implicates the amygdala in Pavlovian fear conditioning. In this study, we examined the contribution of individual amygdaloid nuclei to contextual and auditory fear conditioning in rats. Prior to fear conditioning, rats received a large electrolytic lesion of the amygdala in one hemisphere, and a nucleus-specific neurotoxic lesion in the contralateral hemisphere. Neurotoxic lesions targeted either the lateral nucleus (LA), basolateral and basomedial nuclei (basal nuclei), or central nucleus (CE) of the amygdala. LA and CE lesions attenuated freezing to both contextual and auditory conditional stimuli (CSs). Lesions of the basal nuclei produced deficits in contextual and auditory fear conditioning only when the damage extended into the anterior divisions of the basal nuclei; damage limited to the posterior divisions of the basal nuclei did not significantly impair conditioning to either auditory or contextual CS. These effects were typically not lateralized, although neurotoxic lesions of the posterior divisions of the basal nuclei had greater effects on contextual fear conditioning when the contralateral electrolytic lesion was placed in the right hemisphere. These results indicate that there is significant overlap within the amygdala in the neural pathways mediating fear conditioning to contextual and acoustic CS, and that these forms of learning are not anatomically dissociable at the level of amygdaloid nuclei. PMID- 11390635 TI - Damage to the lateral and central, but not other, amygdaloid nuclei prevents the acquisition of auditory fear conditioning. AB - It is well established that the amygdala plays an essential role in Pavlovian fear conditioning, with the lateral nucleus serving as the interface with sensory systems that transmit the conditioned stimulus and the central nucleus as the link with motor regions that control conditioned fear responses. The lateral nucleus connects with the central nucleus directly and by way of several other amygdala regions, including the basal, accessory basal, and medial nuclei. To determine which of these regions is necessary, and thus whether conditioning requires the direct or one of the indirect intra-amygdala pathways, we made lesions in rats of the lateral, central, basal, accessory basal, and medial nuclei, as well as combined lesions of the basal and accessory basal nuclei and of the entire amygdala. Animals subsequently underwent fear conditioning trials in which an auditory conditioned stimulus was paired with a footshock unconditioned stimulus. Animals that received lesions of the lateral or central nucleus, or of the entire amygdala, were dramatically impaired, whereas the other lesions had little effect. These findings show that only the lateral and central nuclei are necessary for the acquisition of conditioned fear response to an auditory conditioned stimulus. PMID- 11390636 TI - Lesion of the ventral periaqueductal gray reduces conditioned fear but does not change freezing induced by stimulation of the dorsal periaqueductal gray. AB - Previously-reported evidence showed that freezing to a context previously associated with footshock is impaired by lesion of the ventral periaqueductal gray (vPAG). It has also been shown that stepwise increase in the intensity of the electrical stimulation of the dorsal periaqueductal gray (dPAG) produces alertness, then freezing, and finally escape. These aversive responses are mimicked by microinjections of GABA receptor antagonists, such as bicuculline, or blockers of the glutamic acid decarboxylase (GAD), such as semicarbazide, into the dPAG. In this work, we examined whether the expression of these defensive responses could be the result of activation of ventral portion of the periaqueductal gray. Sham- or vPAG electrolytic-lesioned rats were implanted with an electrode in the dPAG for the determination of the thresholds of freezing and escape responses. The vPAG electrolytic lesions were behaviorally verified through a context-conditioned fear paradigm. Results indicated that lesion of the vPAG disrupted conditioned freezing response to contextual cues associated with footshocks but did not change the dPAG electrical stimulation for freezing and escape responses. In a second experiment, lesion of the vPAG also did not change the amount of freezing and escape behavior produced by microinjections of semicarbazide into the dPAG. These findings indicate that freezing and escape defensive responses induced by dPAG stimulation do not depend on the integrity of the vPAG. A discussion on different neural circuitries that might underlie different inhibitory and active defensive behavioral patterns that animals display during threatening situations is presented. PMID- 11390637 TI - Regulation of distinct attractive and aversive mechanisms mediating benzaldehyde chemotaxis in Caenorhabditis elegans. AB - Olfactory-mediated chemotaxis in nematodes provides a relatively simple system to study biological mechanisms of information processing. Analysis of the kinetics of chemotaxis in response to 100% benzaldehyde revealed an initial attractive response that is followed by a strong aversion to the odorant. We show that this behavior is mediated by two genetically separable attraction- and aversion mediating response pathways. The attraction initially dominates behavior but with prolonged exposure habituation leads to a behavioral change, such that the odorant becomes repulsive. This olfactory habituation is susceptible to dishabituation, thereby re-establishing the attractive response to the odorant. Re-examination of the putative olfactory adaptation mutant adp-1(ky20) revealed that the phenotype observed in this line is due to a supersensitivity to a dishabituating stimulus, rather than a defect in the adaptation to odorants per se. A modified benzaldehyde chemotaxis assay was developed and used for the isolation of a mutant with a specific defect in habituation kinetics, expressed as a persistence of the attractive response. PMID- 11390638 TI - Phosphorylation of the RNA polymerase II carboxy-terminal domain by the Bur1 cyclin-dependent kinase. AB - BUR1, which was previously identified by a selection for mutations that have general effects on transcription in Saccharomyces cerevisiae, encodes a cyclin dependent kinase that is essential for viability, but none of its substrates have been identified to date. Using an unbiased biochemical approach, we have identified the carboxy-terminal domain (CTD) of Rpb1, the largest subunit of RNA polymerase II, as a Bur1 substrate. Phosphorylation of Rpb1 by Bur1 is likely to be physiologically relevant, since bur1 mutations interact genetically with rpb1 CTD truncations and with mutations in other genes involved in CTD function. Several genetic interactions are presented, implying a role for Bur1 during transcriptional elongation. These results identify Bur1 as a fourth S. cerevisiae CTD kinase and provide striking functional similarities between Bur1 and metazoan P-TEFb. PMID- 11390639 TI - Eukaryotic initiation factor 4G-poly(A) binding protein interaction is required for poly(A) tail-mediated stimulation of picornavirus internal ribosome entry segment-driven translation but not for X-mediated stimulation of hepatitis C virus translation. AB - Efficient translation of most eukaryotic mRNAs results from synergistic cooperation between the 5' m(7)GpppN cap and the 3' poly(A) tail. In contrast to such mRNAs, the polyadenylated genomic RNAs of picornaviruses are not capped, and translation is initiated internally, driven by an extensive sequence termed IRES (for internal ribosome entry segment). Here we have used our recently described poly(A)-dependent rabbit reticulocyte lysate cell-free translation system to study the role of mRNA polyadenylation in IRES-driven translation. Polyadenylation significantly stimulated translation driven by representatives of each of the three types of picornaviral IRES (poliovirus, encephalomyocarditis virus, and hepatitis A virus, respectively). This did not result from a poly(A) dependent alteration of mRNA stability in our in vitro translation system but was very sensitive to salt concentration. Disruption of the eukaryotic initiation factor 4G-poly(A) binding protein (eIF4G-PABP) interaction or cleavage of eIF4G abolished or severely reduced poly(A) tail-mediated stimulation of picornavirus IRES-driven translation. In contrast, translation driven by the flaviviral hepatitis C virus (HCV) IRES was not stimulated by polyadenylation but rather by the authentic viral RNA 3' end: the highly structured X region. X region-mediated stimulation of HCV IRES activity was not affected by disruption of the eIF4G-PABP interaction. These data demonstrate that the protein-protein interactions required for synergistic cooperativity on capped and polyadenylated cellular mRNAs mediate 3'-end stimulation of picornaviral IRES activity but not HCV IRES activity. Their implications for the picornavirus infectious cycle and for the increasing number of identified cellular IRES-carrying mRNAs are discussed. PMID- 11390640 TI - Pf1, a novel PHD zinc finger protein that links the TLE corepressor to the mSin3A histone deacetylase complex. AB - The mSin3A-histone deacetylase corepressor is a multiprotein complex that is recruited by DNA binding transcriptional repressors. Sin3 has four paired amphipathic alpha helices (PAH1 to -4) that are protein-protein interaction motifs and is the scaffold upon which the complex assembles. We identified a novel mSin3A-interacting protein that has two plant homeodomain (PHD) zinc fingers we term Pf1, for PHD factor one. Pf1 associates with mSin3A in vivo and recruits the mSin3A complex to repress transcription when fused to the DNA binding domain of Gal4. Pf1 interacts with Sin3 through two independent Sin3 interaction domains (SIDs), Pf1SID1 and Pf1SID2. Pf1SID1 binds PAH2, while Pf1SID2 binds PAH1. Pf1SID1 has sequence and structural similarity to the well characterized 13-amino-acid SID of the Mad bHLHZip repressor. Pf1SID2 does not have sequence similarity with either Mad SID or Pf1SID1 and therefore represents a novel Sin3 binding domain. Mutations in a minimal fragment of Pf1 that encompasses Pf1SID1 inhibited mSin3A binding yet only slightly impaired repression when targeted to DNA, implying that Pf1 might interact with other corepressors. We show that Pf1 interacts with a mammalian homolog of the Drosophila Groucho corepressor, transducin-like enhancer (TLE). Pf1 binds TLE in an mSin3A-independent manner and recruits functional TLE complexes to repress transcription. These findings suggest that Pf1 may serve to bridge two global transcription networks, mSin3A and TLE. PMID- 11390641 TI - Loss of annexin A7 leads to alterations in frequency-induced shortening of isolated murine cardiomyocytes. AB - Annexin A7 has been proposed to function in the fusion of vesicles, acting as a Ca(2+) channel and as Ca(2+)-activated GTPase, thus inducing Ca(2+)/GTP-dependent secretory events. To understand the function of annexin A7, we have performed targeted disruption of the Anxa7 gene in mice. Matings between heterozygous mice produced offspring showing a normal Mendelian pattern of inheritance, indicating that the loss of annexin A7 did not interfere with viability in utero. Mice lacking annexin A7 showed no obvious phenotype and were fertile. To assay for exocytosis, insulin secretion from isolated islets of Langerhans was examined. Ca(2+)-induced and cyclic AMP-mediated potentiation of insulin secretion was unchanged in the absence of annexin A7, suggesting that it is not directly implicated in vesicle fusion. Ca(2+) regulation studied in isolated cardiomyocytes, showed that while cells from early embryos displayed intact Ca(2+) homeostasis and expressed all of the components required for excitation contraction coupling, cardiomyocytes from adult Anxa7(-/-) mice exhibited an altered cell shortening-frequency relationship when stimulated with high frequencies. This suggests a function for annexin A7 in electromechanical coupling, probably through Ca(2+) homoeostasis. PMID- 11390643 TI - Early cell cycle box-mediated transcription of CLN3 and SWI4 contributes to the proper timing of the G(1)-to-S transition in budding yeast. AB - The Cln3-Cdc28 kinase is required to activate the Swi4-Swi6 transcription complex which induces CLN1 and CLN2 transcription in late G(1) and drives the transition to S. Cln3 and Swi4 are both rate limiting for G(1) progression, and they are coordinately transcribed to peak at the M/G(1) boundary. Early cell cycle box (ECB) elements, which confer M/G(1)-specific transcription, have been found in both promoters, and elimination of all ECB elements from the CLN3 promoter causes both a loss of periodicity and Cln3-deficient phenotypes, which include an extended G(1) interval and increased cell volume. Mutants lacking the ECB elements in both the CLN3 and SWI4 promoters have low and deregulated levels of CLN transcripts, and the G(1)-to-S transition for these mutants is delayed and highly variable. These observations support the view that the coordinated rise of Cln3 and Swi4 levels mediated by ECB-dependent transcription controls the timing of the G(1)-to-S phase transition. PMID- 11390642 TI - ATR-mediated checkpoint pathways regulate phosphorylation and activation of human Chk1. AB - Chk1 is an evolutionarily conserved protein kinase that regulates cell cycle progression in response to checkpoint activation. In this study, we demonstrated that agents that block DNA replication or cause certain forms of DNA damage induce the phosphorylation of human Chk1. The phosphorylated form of Chk1 possessed higher intrinsic protein kinase activity and eluted more quickly on gel filtration columns. Serines 317 and 345 were identified as sites of phosphorylation in vivo, and ATR (the ATM- and Rad3-related protein kinase) phosphorylated both of these sites in vitro. Furthermore, phosphorylation of Chk1 on serines 317 and 345 in vivo was ATR dependent. Mutants of Chk1 containing alanine in place of serines 317 and 345 were poorly activated in response to replication blocks or genotoxic stress in vivo, were poorly phosphorylated by ATR in vitro, and were not found in faster-eluting fractions by gel filtration. These findings demonstrate that the activation of Chk1 in response to replication blocks and certain forms of genotoxic stress involves phosphorylation of serines 317 and 345. In addition, this study implicates ATR as a direct upstream activator of Chk1 in human cells. PMID- 11390644 TI - Establishment of an oriP replicon is dependent upon an infrequent, epigenetic event. AB - Plasmids containing oriP, the latent origin of replication for Epstein-Barr virus, support efficient replication in selected cell clones when the viral protein EBNA-1 is provided, being lost at a rate of 2 to 4% per cell generation after removal of selection (A. L. Kirchmaier and B. Sugden, J. Virol. 69:1280 1283, 1995; B. Sugden and N. Warren, Mol. Biol. Med. 5:85-94, 1988). We refer to these plasmids as established replicons in that they support efficient DNA synthesis and partitioning each cell cycle. Unexpectedly, we have found that upon introduction of oriP plasmids into a population of EBNA-1-positive cells, oriP plasmids replicate but are lost precipitously from cells during 2 weeks posttransfection (>25% rate of loss per cell generation). Upon investigation of these disparate observations, we have found that only 1 to 10% of cells transfected with an oriP plasmid expressing EBNA-1 and hygromycin phosphotransferase give rise to drug-resistant clones in which the oriP replicon is established. A hereditable alteration in these drug-resistant cell clones, manifested at the genetic or epigenetic level, does not underlie the establishment of oriP, as newly introduced oriP plasmids replicate but are also lost rapidly from these cells. In addition, a genetic alteration in the oriP plasmid is not responsible for establishment, as oriP plasmids isolated from an established cell clone, propagated in Escherichia coli, and reintroduced into EBNA-1-positive cells are likewise established inefficiently. Our findings demonstrate that oriP replicons are not intrinsically stable in EBNA-1-positive cell lines. Rather, the establishment of an oriP replicon is conferred upon the replicon by a stochastic, epigenetic event that occurs infrequently and, therefore, is detected in only a minority of cells. PMID- 11390645 TI - TFIIS enhances transcriptional elongation through an artificial arrest site in vivo. AB - Transcriptional elongation by RNA polymerase II has been well studied in vitro, but understanding of this process in vivo has been limited by the lack of a direct and specific assay. Here, we designed a specific assay for transcriptional elongation in vivo that involves an artificial arrest (ARTAR) site designed from a thermodynamic theory of DNA-dependent transcriptional arrest in vitro. Transcriptional analysis and chromatin immunoprecipitation experiments indicate that the ARTAR site can arrest Pol II in vivo at a position far from the promoter. TFIIS can counteract this arrest, thereby demonstrating that it possesses transcriptional antiarrest activity in vivo. Unexpectedly, the ARTAR site does not function under conditions of high transcriptional activation unless cells are exposed to conditions (6-azauracil or reduced temperature) that are presumed to affect elongation in vivo. Conversely, TFIIS affects gene expression under conditions of high, but not low, transcriptional activation. Our results provide physical evidence for the discontinuity of transcription elongation in vivo, and they suggest that the functional importance of transcriptional arrest sites and TFIIS is strongly influenced by the level of transcriptional activation. PMID- 11390646 TI - Functional characterization of interferon regulatory factor 3a (IRF-3a), an alternative splice isoform of IRF-3. AB - Virus infection of numerous cell types results in the transcriptional induction of a subset of virus- and interferon (IFN)-stimulated genes. The beta IFN (IFN beta) gene is one of these rapidly induced genes; it serves as a fundamental component of the cellular defense response in eliciting potent antiviral, immunomodulatory, and antiproliferative effects. One of the transcription factors involved in the stringent regulation of IFN-beta production following virus infection is interferon regulatory factor (IRF) 3 (IRF-3). We have characterized an alternatively spliced isoform of IRF-3 that we have called IRF-3a. IRF-3a can selectively and potently inhibit virus-induced activation of the IFN-beta promoter. IRF-3a lacks half of the DNA binding domain found in IRF-3 and is unable to bind to the classical IRF binding elements, IFN-stimulated response elements. These studies suggest that IRF-3a may act as a modulator of IRF-3. PMID- 11390647 TI - Docking protein FRS2 links the protein tyrosine kinase RET and its oncogenic forms with the mitogen-activated protein kinase signaling cascade. AB - The receptor tyrosine kinase RET functions as the signal transducing receptor for the GDNF (for "glial cell-derived neurotrophic factors") family of ligands. Mutations in the RET gene were implicated in Hirschsprung disease (HSCR), multiple endocrine neoplasia type 2 (MEN 2), and thyroid carcinomas. In this report we demonstrate that the docking protein FRS2 is tyrosine phosphorylated by ligand-stimulated and by constitutively activated oncogenic forms of RET. Complex formation between RET and FRS2 is mediated by binding of the phosphotyrosine binding domain of FRS2 to pY1062, a residue in RET that also functions as a binding site for Shc. However, overexpression of FRS2 but not Shc potentiates mitogen-activated protein (MAP) kinase activation by RET oncoproteins. We demonstrate that oncogenic RET-PTC proteins are associated with FRS2 constitutively, leading to tyrosine phosphorylation of FRS2, MAP kinase stimulation, and cell proliferation. However, loss-of-function HSCR-associated RET mutants exhibit impaired FRS2 binding and reduced MAP kinase activation. These experiments demonstrate that FRS2 couples both ligand-regulated and oncogenic forms of RET, with the MAP kinase signaling cascade as part of the response of RET under normal biological conditions and pathological conditions, such as MEN 2 and papillary thyroid carcinomas. PMID- 11390648 TI - Impaired activity of the extraneuronal monoamine transporter system known as uptake-2 in Orct3/Slc22a3-deficient mice. AB - Two uptake systems that control the extracellular concentrations of released monoamine neurotransmitters such as noradrenaline and adrenaline have been described. Uptake-1 is present at presynaptic nerve endings, whereas uptake-2 is extraneuronal and has been identified in myocardium and vascular and nonvascular smooth muscle cells. The gene encoding the uptake-2 transporter has recently been identified in humans (EMT), rats (OCT3), and mice (Orct3/Slc22a3). To generate an in vivo model for uptake-2, we have inactivated the mouse Orct3 gene. Homozygous mutant mice are viable and fertile with no obvious physiological defect and also show no significant imbalance of noradrenaline or dopamine. However, Orct3-null mice show an impaired uptake-2 activity as measured by accumulation of intravenously administered [(3)H]MPP(+) (1-methyl-4-phenylpyridinium). A 72% reduction in MPP(+) levels was measured in hearts of both male and female Orct3 mutant mice. No significant differences between wild-type and mutant mice were found in any other adult organ or in plasma. When [(3)H]MPP(+) was injected into pregnant females, a threefold-reduced MPP(+) accumulation was observed in homozygous mutant embryos but not in their placentas or amniotic fluid. These data show that Orct3 is the principal component for uptake-2 function in the adult heart and identify the placenta as a novel site of action of uptake-2 that acts at the fetoplacental interface. PMID- 11390649 TI - Intrasteric inhibition of ATP binding is not required to prevent unregulated autophosphorylation or signaling by the insulin receptor. AB - Receptor tyrosine kinases may use intrasteric inhibition to suppress autophosphorylation prior to growth factor stimulation. To test this hypothesis we made an Asp1161Ala mutant in the activation loop that relieved intrasteric inhibition of the unphosphorylated insulin receptor (IR) and its recombinant cytoplasmic kinase domain (IRKD) without affecting the activated state. Solution studies with the unphosphorylated mutant IRKD demonstrated conformational changes and greater catalytic efficiency from a 10-fold increase in k(cat) and a 15-fold lower K(m ATP) although K(m peptide) was unchanged. Kinetic parameters of the autophosphorylated mutant and wild-type kinase domains were virtually identical. The Asp1161Ala mutation increased the rate of in vitro autophosphorylation of the IRKD or IR at low ATP concentrations and in the absence of insulin. However, saturation with ATP (for the IRKD) or the presence of insulin (for the IR) yielded equivalent rates of autophosphorylation for mutant versus wild-type kinases. Despite a biochemically more active kinase domain, the mutant IR expressed in C2C12 myoblasts was not constitutively autophosphorylated. However, it displayed a 2.5-fold-lower 50% effective concentration for insulin stimulation of autophosphorylation and was dephosphorylated more slowly following withdrawal of insulin than wild-type IR. In tests of the regulation of the unphosphorylated basal state, these results demonstrate that neither intrasteric inhibition against ATP binding nor suppression of kinase activity is required to prevent premature autophosphorylation of the IR. Finally, the lower rate of dephosphorylation suggests invariant residues of the activation loop such as Asp1161 may function at multiple junctures in cellular regulation of receptor tyrosine kinases. PMID- 11390651 TI - The yeast hnRNP-Like proteins Yra1p and Yra2p participate in mRNA export through interaction with Mex67p. AB - Yra1p is an essential nuclear protein which belongs to the evolutionarily conserved REF (RNA and export factor binding proteins) family of hnRNP-like proteins. Yra1p contributes to mRNA export in vivo and directly interacts with RNA and the shuttling mRNP export receptor Mex67p in vitro. Here we describe a second nonessential Saccharomyces cerevisiae family member, called Yra2p, which is able to complement a YRA1 deletion when overexpressed. Like other REF proteins, Yra1p and Yra2p consist of two highly conserved N- and C-terminal boxes and a central RNP-like RNA-binding domain (RBD). These conserved regions are separated by two more variable regions, N-vr and C-vr. Surprisingly, the deletion of a single conserved box or the deletion of the RBD in Yra1p does not affect viability. Consistently, neither the conserved N and C boxes nor the RBD is required for Mex67p and RNA binding in vitro. Instead, the N-vr and C-vr regions both interact with Mex67p and RNA. We further show that Yra1 deletion mutants which poorly interact with Mex67p in vitro affect the association of Mex67p with mRNP complexes in vivo and are paralleled by poly(A)(+) RNA export defects. These observations support the idea that Yra1p promotes mRNA export by facilitating the recruitment of Mex67p to the mRNP. PMID- 11390650 TI - Identification of a phospholipase C-gamma1 (PLC-gamma1) SH3 domain-binding site in SLP-76 required for T-cell receptor-mediated activation of PLC-gamma1 and NFAT. AB - SLP-76 is an adapter protein required for T-cell receptor (TCR) signaling. In particular, TCR-induced tyrosine phosphorylation and activation of phospholipase C-gamma1 (PLC-gamma1), and the resultant TCR-inducible gene expression, depend on SLP-76. Nonetheless, the mechanisms by which SLP-76 mediates PLC-gamma1 activation are not well understood. We now demonstrate that SLP-76 directly interacts with the Src homology 3 (SH3) domain of PLC-gamma1. Structure-function analysis of SLP-76 revealed that each of the previously defined protein-protein interaction domains can be individually deleted without completely disrupting SLP 76 function. Additional deletion mutations revealed a new, 67-amino-acid functional domain within the proline-rich region of SLP-76, which we have termed the P-1 domain. The P-1 domain mediates a constitutive interaction of SLP-76 with the SH3 domain of PLC-gamma1 and is required for TCR-mediated activation of Erk, PLC-gamma1, and NFAT (nuclear factor of activated T cells). The adjacent Gads binding domain of SLP-76, also within the proline-rich region, mediates inducible recruitment of SLP-76 to a PLC-gamma1-containing complex via the recruitment of both PLC-gamma1 and Gads to another cell-type-specific adapter, LAT. Thus, TCR induced activation of PLC-gamma1 entails the binding of PLC-gamma1 to both LAT and SLP-76, a finding that may underlie the requirement for both LAT and SLP-76 to mediate the optimal activation of PLC-gamma1. PMID- 11390652 TI - New function of CDC13 in positive telomere length regulation. AB - Two roles for the Saccharomyces cerevisiae Cdc13 protein at the telomere have previously been characterized: it recruits telomerase to the telomere and protects chromosome ends from degradation. In a synthetic lethality screen with YKU70, the 70-kDa subunit of the telomere-associated Yku heterodimer, we identified a new mutation in CDC13, cdc13-4, that points toward an additional regulatory function of CDC13. Although CDC13 is an essential telomerase component in vivo, no replicative senescence can be observed in cdc13-4 cells. Telomeres of cdc13-4 mutants shorten for about 150 generations until they reach a stable level. Thus, in cdc13-4 mutants, telomerase seems to be inhibited at normal telomere length but fully active at short telomeres. Furthermore, chromosome end structure remains protected in cdc13-4 mutants. Progressive telomere shortening to a steady-state level has also been described for mutants of the positive telomere length regulator TEL1. Strikingly, cdc13-4/tel1Delta double mutants display shorter telomeres than either single mutant after 125 generations and a significant amplification of Y' elements after 225 generations. Therefore CDC13, TEL1, and the Yku heterodimer seem to represent distinct pathways in telomere length maintenance. Whereas several CDC13 mutants have been reported to display elongated telomeres indicating that Cdc13p functions in negative telomere length control, we report a new mutation leading to shortened and eventually stable telomeres. Therefore we discuss a key role of CDC13 not only in telomerase recruitment but also in regulating telomerase access, which might be modulated by protein-protein interactions acting as inhibitors or activators of telomerase activity. PMID- 11390653 TI - Evidence of p53-dependent cross-talk between ribosome biogenesis and the cell cycle: effects of nucleolar protein Bop1 on G(1)/S transition. AB - Bop1 is a novel nucleolar protein involved in rRNA processing and ribosome assembly. We have previously shown that expression of Bop1Delta, an amino terminally truncated Bop1 that acts as a dominant negative mutant in mouse cells, results in inhibition of 28S and 5.8S rRNA formation and deficiency of newly synthesized 60S ribosomal subunits (Z. Strezoska, D. G. Pestov, and L. F. Lau, Mol. Cell. Biol. 20:5516-5528, 2000). Perturbation of Bop1 activities by Bop1Delta also induces a powerful yet reversible cell cycle arrest in 3T3 fibroblasts. In the present study, we show that asynchronously growing cells are arrested by Bop1Delta in a highly concerted fashion in the G(1) phase. Kinase activities of the G(1)-specific Cdk2 and Cdk4 complexes were downregulated in cells expressing Bop1Delta, whereas levels of the Cdk inhibitors p21 and p27 were concomitantly increased. The cells also displayed lack of hyperphosphorylation of retinoblastoma protein (pRb) and decreased expression of cyclin A, indicating their inability to progress through the restriction point. Inactivation of functional p53 abrogated this Bop1Delta-induced cell cycle arrest but did not restore normal rRNA processing. These findings show that deficiencies in ribosome synthesis can be uncoupled from cell cycle arrest and reveal a new role for the p53 pathway as a mediator of the signaling link between ribosome biogenesis and the cell cycle. We propose that aberrant rRNA processing and/or ribosome biogenesis may cause "nucleolar stress," leading to cell cycle arrest in a p53 dependent manner. PMID- 11390654 TI - Functional organization of single and paired V(D)J cleavage complexes. AB - RAG-1 and RAG-2 initiate V(D)J recombination by binding to specific recognition sequences (RSS) and then cleave the DNA in two steps: nicking and hairpin formation. Recent work has established that a dimer of RAG-1 and either one or two monomers of RAG-2 bind to a single RSS, but the enzymatic contributions of the RAG molecules within this nucleoprotein complex and its functional organization have not been elucidated. Using heterodimeric protein preparations containing both wild-type and catalytically deficient RAG-1 molecules, we found that one active monomer is sufficient for both nicking and hairpin formation at a single RSS, demonstrating that a single active site can carry out both cleavage steps. Furthermore, the mutant heterodimers efficiently cleaved both RSS in a synaptic complex. These results strongly suggest that two RAG-1 dimers are responsible for RSS cleavage in a synaptic complex, with one monomer of each dimer catalyzing both nicking and hairpin formation at each RSS. PMID- 11390655 TI - Her4 mediates ligand-dependent antiproliferative and differentiation responses in human breast cancer cells. AB - The function of the epidermal growth factor receptor (EGFR) family member HER4 remains unclear because its activating ligand, heregulin, results in either proliferation or differentiation. This variable response may stem from the range of signals generated by HER4 homodimers versus heterodimeric complexes with other EGFR family members. The ratio of homo- and heterodimeric complexes may be influenced both by a cell's EGFR family member expression profile and by the ligand or even ligand isoform used. To define the role of HER4 in mediating antiproliferative and differentiation responses, human breast cancer cell lines were screened for responses to heregulin. Only cells that expressed HER4 exhibited heregulin-dependent antiproliferative responses. In-depth studies of one line, SUM44, demonstrated that the antiproliferative and differentiation responses correlated with HER4 activation and were abolished by stable expression of a kinase-inactive HER4. HB-EGF, a HER4-specific ligand in this EGFR-negative cell line, also induced an antiproliferative response. Moreover, introduction and stable expression of HER4 in HER4-negative SUM102 cells resulted in the acquisition of a heregulin-dependent antiproliferative response, associated with increases in markers of differentiation. The role of HER2 in these heregulin dependent responses was examined through elimination of cell surface HER2 signaling by stable expression of a single-chain anti-HER2 antibody that sequestered HER2 in the endoplasmic reticulum. In the cell lines with either endogenously (SUM44) or exogenously (SUM102) expressed HER4, elimination of HER2 did not alter HER4-dependent decreases in cell growth. These results suggest that HER4 is both necessary and sufficient to trigger an antiproliferative response in human breast cancer cells. PMID- 11390657 TI - Molecular cloning of ILP-2, a novel member of the inhibitor of apoptosis protein family. AB - Inhibitor of apoptosis protein (IAP)-like protein-1 (ILP-1) (also known as X linked IAP [XIAP] and mammalian IAP homolog A [MIHA]) is a potent inhibitor of apoptosis and exerts its effects, at least in part, by the direct association with and inhibition of specific caspases. Here, we describe the molecular cloning and characterization of a human gene related to ILP-1, termed ILP-2. Despite high homology to ILP-1, ILP-2 is encoded by a distinct gene, which in normal tissues is expressed solely in testis. In contrast to ILP-1, overexpression of ILP-2 had no protective effect on apoptosis mediated by Fas (also known as CD95) or tumor necrosis factor. However, ILP-2 potently inhibited apoptosis induced by overexpression of Bax or by coexpression of caspase 9 with Apaf-1, and preincubation of cytosolic extracts with ILP-2 abrogated caspase activation in vitro. A processed form of caspase 9 could be coprecipitated with ILP-2 from cells, suggesting a physical interaction between ILP-2 and caspase 9. Thus, ILP-2 is a novel IAP family member with restricted specificity for caspase 9. PMID- 11390656 TI - In vivo action of the HRD ubiquitin ligase complex: mechanisms of endoplasmic reticulum quality control and sterol regulation. AB - Ubiquitination is used to target both normal proteins for specific regulated degradation and misfolded proteins for purposes of quality control destruction. Ubiquitin ligases, or E3 proteins, promote ubiquitination by effecting the specific transfer of ubiquitin from the correct ubiquitin-conjugating enzyme, or E2 protein, to the target substrate. Substrate specificity is usually determined by specific sequence determinants, or degrons, in the target substrate that are recognized by the ubiquitin ligase. In quality control, however, a potentially vast collection of proteins with characteristic hallmarks of misfolding or misassembly are targeted with high specificity despite the lack of any sequence similarity between substrates. In order to understand the mechanisms of quality control ubiquitination, we have focused our attention on the first characterized quality control ubiquitin ligase, the HRD complex, which is responsible for the endoplasmic reticulum (ER)-associated degradation (ERAD) of numerous ER-resident proteins. Using an in vivo cross-linking assay, we directly examined the association of the separate HRD complex components with various ERAD substrates. We have discovered that the HRD ubiquitin ligase complex associates with both ERAD substrates and stable proteins, but only mediates ubiquitin-conjugating enzyme association with ERAD substrates. Our studies with the sterol pathway regulated ERAD substrate Hmg2p, an isozyme of the yeast cholesterol biosynthetic enzyme HMG-coenzyme A reductase (HMGR), indicated that the HRD complex discerns between a degradation-competent "misfolded" state and a stable, tightly folded state. Thus, it appears that the physiologically regulated, HRD-dependent degradation of HMGR is effected by a programmed structural transition from a stable protein to a quality control substrate. PMID- 11390658 TI - RAG transposase can capture and commit to target DNA before or after donor cleavage. AB - The discovery that the V(D)J recombinase functions as a transposase in vitro suggests that transposition by this system might be a potent source of genomic instability. To gain insight into the mechanisms that regulate transposition, we investigated a phenomenon termed target commitment that reflects a functional association between the RAG transposase and the target DNA. We found that the V(D)J recombinase is quite promiscuous, forming productive complexes with target DNA both before and after donor cleavage, and our data indicate that the rate limiting step for transposition occurs after target capture. Formation of stable target capture complexes depends upon the presence of active-site metal binding residues (the DDE motif), suggesting that active-site amino acids in RAG-1 are critical for target capture. The ability of the RAG transposase to commit to target prior to cleavage may result in a preference for transposition into nearby targets, such as immunoglobulin and T-cell receptor loci. This could bias transposition toward relatively "safe" regions of the genome. A preference for localized transposition may also have influenced the evolution of the antigen receptor loci. PMID- 11390659 TI - Essential roles of Snf5p in Snf-Swi chromatin remodeling in vivo. AB - Snf-Swi, the prototypical ATP-dependent nucleosome-remodeling complex, regulates transcription of a subset of yeast genes. With the exception of Snf2p, the ATPase subunit, the functions of the other components are unknown. We have investigated the role of the conserved Snf-Swi core subunit Snf5p through characterization of two conditional snf5 mutants. The mutants contain single amino acid alterations of invariant or conserved residues that abolish Snf-Swi-dependent transcription by distinct mechanisms. One mutation impairs Snf-Swi assembly and, consequently, its stable association with a target promoter. The other blocks a postrecruitment catalytic remodeling step. These findings suggest that Snf5p coordinates the assembly and nucleosome-remodeling activities of Snf-Swi. PMID- 11390660 TI - Identification of a peroxisomal ATP carrier required for medium-chain fatty acid beta-oxidation and normal peroxisome proliferation in Saccharomyces cerevisiae. AB - We have characterized the role of YPR128cp, the orthologue of human PMP34, in fatty acid metabolism and peroxisomal proliferation in Saccharomyces cerevisiae. YPR128cp belongs to the mitochondrial carrier family (MCF) of solute transporters and is localized in the peroxisomal membrane. Disruption of the YPR128c gene results in impaired growth of the yeast with the medium-chain fatty acid (MCFA) laurate as a single carbon source, whereas normal growth was observed with the long-chain fatty acid (LCFA) oleate. MCFA but not LCFA beta-oxidation activity was markedly reduced in intact ypr128cDelta mutant cells compared to intact wild type cells, but comparable activities were found in the corresponding lysates. These results imply that a transport step specific for MCFA beta-oxidation is impaired in ypr128cDelta cells. Since MCFA beta-oxidation in peroxisomes requires both ATP and CoASH for activation of the MCFAs into their corresponding coenzyme A esters, we studied whether YPR128cp is an ATP carrier. For this purpose we have used firefly luciferase targeted to peroxisomes to measure ATP consumption inside peroxisomes. We show that peroxisomal luciferase activity was strongly reduced in intact ypr128cDelta mutant cells compared to wild-type cells but comparable in lysates of both cell strains. We conclude that YPR128cp most likely mediates the transport of ATP across the peroxisomal membrane. PMID- 11390661 TI - The polycomb-group gene Ezh2 is required for early mouse development. AB - Polycomb-group (Pc-G) genes are required for the stable repression of the homeotic selector genes and other developmentally regulated genes, presumably through the modulation of chromatin domains. Among the Drosophila Pc-G genes, Enhancer of zeste [E(z)] merits special consideration since it represents one of the Pc-G genes most conserved through evolution. In addition, the E(Z) protein family contains the SET domain, which has recently been linked with histone methyltransferase (HMTase) activity. Although E(Z)-related proteins have not (yet) been directly associated with HMTase activity, mammalian Ezh2 is a member of a histone deacetylase complex. To investigate its in vivo function, we generated mice deficient for Ezh2. The Ezh2 null mutation results in lethality at early stages of mouse development. Ezh2 mutant mice either cease developing after implantation or initiate but fail to complete gastrulation. Moreover, Ezh2 deficient blastocysts display an impaired potential for outgrowth, preventing the establishment of Ezh2-null embryonic stem cells. Interestingly, Ezh2 is up regulated upon fertilization and remains highly expressed at the preimplantation stages of mouse development. Together, these data suggest an essential role for Ezh2 during early mouse development and genetically link Ezh2 with eed and YY1, the only other early-acting Pc-G genes. PMID- 11390662 TI - A human protein with sequence similarity to Drosophila mastermind coordinates the nuclear form of notch and a CSL protein to build a transcriptional activator complex on target promoters. AB - Mastermind (Mam) has been implicated as an important positive regulator of the Notch signaling pathway by genetic studies using Drosophila melanogaster. Here we describe a biochemical mechanism of action of Mam within the Notch signaling pathway. Expression of a human sequence related to Drosophila Mam (hMam-1) in mammalian cells augments induction of Hairy Enhancer of split (HES) promoters by Notch signaling. hMam-1 stabilizes and participates in the DNA binding complex of the intracellular domain of human Notch1 and a CSL protein. Truncated versions of hMam-1 that can maintain an association with the complex behave in a dominant negative fashion and depress transactivation. Furthermore, Drosophila Mam forms a similar complex with the intracellular domain of Drosophila Notch and Drosophila CSL protein during activation of Enhancer of split, the Drosophila counterpart of HES. These results indicate that Mam is an essential component of the transcriptional apparatus of Notch signaling. PMID- 11390664 TI - Complex functions of AP-1 transcription factors in differentiation and survival of PC12 cells. AB - c-Jun activation by mitogen-activated protein kinases has been implicated in various cellular signal responses. We investigated how JNK and c-Jun contribute to neuronal differentiation, cell survival, and apoptosis. In differentiated PC12 cells, JNK signaling can induce apoptosis and c-Jun mediates this response. In contrast, we show that in PC12 cells that are not yet differentiated, the AP-1 family member ATF-2 and not c-Jun acts as an executor of apoptosis. In this context c-Jun expression protects against apoptosis and triggers neurite formation. Thus, c-Jun has opposite functions before and after neuronal differentiation. These findings suggest a model in which the balance between ATF 2 and Jun activity in PC12 cells governs the choice between differentiation towards a neuronal fate and an apoptotic program. Further analysis of c-Jun mutants showed that the differentiation response requires functional dimerization and DNA-binding domains and that it is stimulated by phosphorylation in the transactivation domain. In contrast, c-Jun mutants incompetent for DNA binding or dimerization and also mutants lacking JNK binding and phosphorylation sites that cannot elicit neuronal differentiation efficiently protect PC12 cells from apoptosis. Hence, the protective role of c-Jun appears to be mediated by an unconventional mechanism that is separable from its function as a classical AP-1 transcription factor. PMID- 11390663 TI - Transcriptional profiling shows that Gcn4p is a master regulator of gene expression during amino acid starvation in yeast. AB - Starvation for amino acids induces Gcn4p, a transcriptional activator of amino acid biosynthetic genes in Saccharomyces cerevisiae. In an effort to identify all genes regulated by Gcn4p during amino acid starvation, we performed cDNA microarray analysis. Data from 21 pairs of hybridization experiments using two different strains derived from S288c revealed that more than 1,000 genes were induced, and a similar number were repressed, by a factor of 2 or more in response to histidine starvation imposed by 3-aminotriazole (3AT). Profiling of a gcn4Delta strain and a constitutively induced mutant showed that Gcn4p is required for the full induction by 3AT of at least 539 genes, termed Gcn4p targets. Genes in every amino acid biosynthetic pathway except cysteine and genes encoding amino acid precursors, vitamin biosynthetic enzymes, peroxisomal components, mitochondrial carrier proteins, and autophagy proteins were all identified as Gcn4p targets. Unexpectedly, genes involved in amino acid biosynthesis represent only a quarter of the Gcn4p target genes. Gcn4p also activates genes involved in glycogen homeostasis, and mutant analysis showed that Gcn4p suppresses glycogen levels in amino acid-starved cells. Numerous genes encoding protein kinases and transcription factors were identified as targets, suggesting that Gcn4p is a master regulator of gene expression. Interestingly, expression profiles for 3AT and the alkylating agent methyl methanesulfonate (MMS) overlapped extensively, and MMS induced GCN4 translation. Thus, the broad transcriptional response evoked by Gcn4p is produced by diverse stress conditions. Finally, profiling of a gcn4Delta mutant uncovered an alternative induction pathway operating at many Gcn4p target genes in histidine-starved cells. PMID- 11390665 TI - Use of suppressor mutants to probe the function of estrogen receptor-p160 coactivator interactions. AB - Estrogen-dependent recruitment of coactivators by estrogen receptor alpha (ERalpha) represents a crucial step in the transcriptional activation of target genes. However, studies of the function of individual coactivators has been hindered by the presence of endogenous coactivators, many of which are potentially recruited in the presence of agonist via a common mechanism. To circumvent this problem, we have generated second-site suppressor mutations in the nuclear receptor interaction domain of p160 coactivators which rescue their binding to a transcriptionally defective ERalpha that is refractory to wild-type coactivators. Analysis of these altered-specificity receptor-coactivator combinations, in the absence of interference from endogenous coregulators, indicated that estrogen-dependent transcription from reporter genes is critically dependent on direct recruitment of a p160 coactivator in mammalian cells and that the three p160 family members serve functionally redundant roles. Furthermore, our results suggest that such a change-of-specificity mutation may act as a transposable protein-protein interaction module which provides a novel tool with which to dissect the functional roles of other nuclear receptor coregulators at the cellular level. PMID- 11390666 TI - Nkx2.5 and Nkx2.6, homologs of Drosophila tinman, are required for development of the pharynx. AB - Nkx2.5 and Nkx2.6 are murine homologs of Drosophila tinman. Their genes are expressed in the ventral region of the pharynx at early stages of embryogenesis. However, no abnormalities in the pharynges of embryos with mutations in either Nkx2.5 or Nkx2.6 have been reported. To examine the function of Nkx2.5 and Nkx2.6 in the formation of the pharynx, we generated and analyzed Nkx2.5 and Nkx2.6 double-mutant mice. Interestingly, in the double-mutant embryos, the pharynx did not form properly. Pharyngeal endodermal cells were largely missing, and the mutant pharynx was markedly dilated. Moreover, we observed enhanced apoptosis and reduced proliferation in pharyngeal endodermal cells of the double-mutant embryos. These results demonstrated a critical role of the NK-2 homeobox genes in the differentiation, proliferation, and survival of pharyngeal endodermal cells. Furthermore, the development of the atrium was less advanced in the double-mutant embryos, indicating that these two genes are essential for both pharyngeal and cardiac development. PMID- 11390667 TI - Mice lacking the Nkx6.2 (Gtx) homeodomain transcription factor develop and reproduce normally. AB - The Nkx homeobox genes are expressed in a variety of developing tissues and have been implicated in controlling tissue patterning and cell differentiation. Expression of Nkx6.2 (Gtx) was previously observed in the embryonic neural tube, testis, and differentiating oligodendrocytes. To investigate the role of Nkx6.2 in the control of cell specification and differentiation, we generated mice with null mutations in Nkx6.2 using the standard gene targeting approach. Null mutant mice were viable and fertile without apparent histological and immunohistochemical changes in the central nervous systems and testis. The absence of detectable phenotypes suggests a redundant function of Nkx6.2 in mouse development. PMID- 11390668 TI - Ligand-mediated assembly and real-time cellular dynamics of estrogen receptor alpha-coactivator complexes in living cells. AB - Studies with live cells demonstrate that agonist and antagonist rapidly (within minutes) modulate the subnuclear dynamics of estrogen receptor alpha (ER) and steroid receptor coactivator 1 (SRC-1). A functional cyan fluorescent protein (CFP)-tagged lac repressor-ER chimera (CFP-LacER) was used in live cells to discretely immobilize ER on stably integrated lac operator arrays to study recruitment of yellow fluorescent protein (YFP)-steroid receptor coactivators (YFP-SRC-1 and YFP-CREB binding protein [CBP]). In the absence of ligand, YFP-SRC 1 is found dispersed throughout the nucleoplasm, with a surprisingly high accumulation on the CFP-LacER arrays. Agonist addition results in the rapid (within minutes) recruitment of nucleoplasmic YFP-SRC-1, while antagonist additions diminish YFP-SRC-1-CFP-LacER associations. Less ligand-independent colocalization is observed with CFP-LacER and YFP-CBP, but agonist-induced recruitment occurs within minutes. The agonist-induced recruitment of coactivators requires helix 12 and critical residues in the ER-SRC-1 interaction surface, but not the F, AF-1, or DNA binding domains. Fluorescence recovery after photobleaching indicates that YFP-SRC-1, YFP-CBP, and CFP-LacER complexes undergo rapid (within seconds) molecular exchange even in the presence of an agonist. Taken together, these data suggest a dynamic view of receptor-coregulator interactions that is now amenable to real-time study in living cells. PMID- 11390670 TI - The discs of Leptothrix discophora: lost for 89 years? PMID- 11390669 TI - Human pex19p binds peroxisomal integral membrane proteins at regions distinct from their sorting sequences. AB - The molecular machinery underlying peroxisomal membrane biogenesis is not well understood. The observation that cells deficient in the peroxins Pex3p, Pex16p, and Pex19p lack peroxisomal membrane structures suggests that these molecules are involved in the initial stages of peroxisomal membrane formation. Pex19p, a predominantly cytosolic protein that can be farnesylated, binds multiple peroxisomal integral membrane proteins, and it has been suggested that it functions as a soluble receptor for the targeting of peroxisomal membrane proteins (PMPs) to the peroxisome. An alternative view proposes that Pex19p functions as a chaperone at the peroxisomal membrane. Here, we show that the peroxisomal sorting determinants and the Pex19p-binding domains of a number of PMPs are distinct entities. In addition, we extend the list of peroxins with which human Pex19p interacts to include the PMP Pex16p and show that Pex19p's CaaX prenylation motif is an important determinant in the affinity of Pex19p for Pex10p, Pex11pbeta, Pex12p, and Pex13p. PMID- 11390671 TI - Ustilago maydis--a valuable model system for the study of fungal dimorphism and virulence. PMID- 11390672 TI - Microarrays for microbiologists. PMID- 11390673 TI - A novel filamentous Bacillus sp., strain NAF001, forming endospores and budding cells. AB - A novel filamentous bacterium, strain NAF001, was isolated from suspended water of a domestic wastewater treatment tank. It formed an extremely long filamentous trichome and produced endospores. It formed spore-like resting cells (SLRCs) which were heat-resistant. SLRCs grew by budding to form short filaments resembling the gonidia of filamentous bacteria such as LEUCOTHRIX: This is the first report of a Bacillus species that exhibits budding growth. The filamentous form was neither restricted to any particular growth stage nor dependent on cultural conditions. Phylogenetic analysis of the 16S rRNA gene revealed that this isolate was a member of the genus Bacillus, with no close relatives at the species level (sequence similarity <95.3%). Strain NAF001 thus probably belongs to a new and novel species of Bacillus. PMID- 11390674 TI - Visualization of Borrelia burgdorferi sensu lato by fluorescence in situ hybridization (FISH) on whole-body sections of Ixodes ricinus ticks and gerbil skin biopsies. AB - The objective of this study was to visualize borreliae directly in whole-body sections of Ixodes ricinus by fluorescence in situ hybridization (FISH). Borrelia afzelii mono-infected or Borrelia burgdorferi sensu stricto (ss)/B. afzelii double-infected nymphs were fixed, embedded in cold polymerizing resin and sectioned. The same sample processing was applied to skin biopsies taken from a Mongolian gerbil after an infectious tick-bite. FISH was carried out using 16S rRNA-directed, fluorescence-labelled oligonucleotide probes specific for the genus Borrelia and specific within the group of Lyme borreliosis-associated genospecies B. afzelii, B. burgdorferi ss, Borrelia garinii and Borrelia valaisiana. Sensitivity and specificity of the newly designed probes were evaluated using PCR, dot-blot hybridizations and FISH. Despite significant autofluorescence of certain tick tissues (which allowed good histological orientation within the sections), borreliae showing the typical spirochaetal morphotype were clearly visible in five out of six putatively infected ticks. These findings were confirmed by electron microscopy of ticks from the same infected batch as used for FISH. Attempts to produce ticks infected by two different Borrelia genospecies were not successful. FISH on whole-body sections of resin-embedded ticks offers the possibility of visualizing and identifying borreliae within tick tissues. This technique has great potential for the investigation of the transmission of bacteria or other micro-organisms by arthropod vectors. Furthermore, clear visualization of single spirochaetes distributed along subcutaneous fat cell membranes in gerbil skin biopsies suggests that FISH might also be suitable for the detection of borreliae in clinical tissue specimens. PMID- 11390675 TI - Cell cycle control of septin ring dynamics in the budding yeast. AB - Septins constitute a cytoskeletal structure that is conserved in eukaryotes. In Saccharomyces cerevisiae, the Cdc3, Cdc10, Cdc11, Cdc12 and Shs1/Sep7 septins assemble as a ring that marks the cytokinetic plane throughout the budding cycle. This structure participates in different aspects of morphogenesis, such as selection of cell polarity, localization of chitin synthesis, the switch from hyperpolar to isotropic bud growth after bud emergence and the spatial regulation of septation. The septin cytoskeleton assembles at the pre-bud site before bud emergence, remains there during bud growth and duplicates at late mitosis eventually disappearing after cell separation. Using a septin-GFP fusion and time lapse confocal microscopy, we have determined that septin dynamics are maintained in budding zygotes and during unipolar synchronous growth in pseudohyphae. By means of specific cell cycle arrests and deregulation of cell cycle controls we show that septin assembly is dependent on G1 cyclin/Cdc28-mediated cell cycle signals and that the small GTPase Cdc42, but not Rho1, are essential for this event. However, during bud growth, the septin ring shapes a bud-neck-spanning structure that is unaffected by failures in the regulation of mitosis, such as activation of the DNA repair or spindle assembly checkpoints or inactivation of the anaphase-promoting complex (APC). At the end of the cell cycle, the splitting of the ring into two independent structures depends on the function of the mitotic exit network in which the protein phosphatase Cdc14 participates. Our data support a role of cell cycle control mechanisms in the regulation of septin dynamics to accurately coordinate morphogenesis throughout the budding process in yeast. PMID- 11390676 TI - Identification of lipopolysaccharide O antigen synthesis genes required for attachment of the S-layer of Caulobacter crescentus. AB - The outer surface of Caulobacter crescentus consists of a two-dimensional crystalline protein lattice layer (S-layer). A fraction of the LPS has an O antigen polymer attached to the core to form a 'smooth' LPS (S-LPS), which is required for attachment of the protein S-layer to the outer-membrane surface. A method to screen for strains defective in LPS production, based on loss of S layer attachment, was developed and applied to libraries of transposon-generated mutants. Eighteen distinct insertions were found with transposon interruptions in genes affecting S-LPS production, 12 of which were located near the S-layer subunit protein gene, rsaA, and its transporter genes. Sequence adjacent to transposon insertion points was determined and used to search a C. crescentus genome database. Twelve ORFs likely to be involved in S-LPS synthesis were identified. Seven of the predicted ORFs were linked to rsaA. Six of the putative genes had identity with proteins involved in synthesis of sugar residues, including five predicted to make perosamine. The remaining six ORFs were similar to glycosyltransferases involved in forming linkages between sugar residues in the O antigen, while one may be a transcription repressor. Other chemical and preliminary proton NMR studies of the S-LPS O antigen indicate that it contains an N-acetylated 4,6-dideoxy-4-aminohexose, but is not assembled as a simple, uniform homopolymer, consisting of several different linkages between sugar residues. The ORFs described here include homologues of all the enzymes involved in the synthesis of N-acetylperosamine, a 4,6-dideoxy-4-aminohexose. Overall, the data are consistent with the hypothesis that the O antigen of C. crescentus S-LPS consists primarily of N-acetylperosamine residues polymerized with multiple anomeric linkages. PMID- 11390677 TI - Metabolic flux in cellulose batch and cellulose-fed continuous cultures of Clostridium cellulolyticum in response to acidic environment. AB - Clostridium cellulolyticum, a nonruminal cellulolytic mesophilic bacterium, was grown in batch and continuous cultures on cellulose using a chemically defined medium. In batch culture with unregulated pH, less cellulose degradation and higher accumulation of soluble glucides were obtained compared to a culture with the pH controlled at 7.2. The gain in cellulose degradation achieved with pH control was offset by catabolite production rather than soluble sugar accumulation. The pH-controlled condition improved biomass, ethanol and acetate production, whereas maximum lactate and extracellular pyruvate concentrations were lower than in the non-pH-controlled condition. In a cellulose-fed chemostat at constant dilution rate and pH values ranging from 7.4 to 6.2, maximum cell density was obtained at pH 7.0. Environmental acidification chiefly influenced biomass formation, since at pH 6.4 the dry weight of cells was more than fourfold lower compared to that at pH 7.0, whereas the specific rate of cellulose assimilation decreased only from 11.74 to 10.13 milliequivalents of carbon (g cells)(-1) h(-1). The molar growth yield and the energetic growth yield did not decline as pH was lowered, and an abrupt transition to washout was observed. Decreasing the pH induced a shift from an acetate-ethanol fermentation to a lactate-ethanol fermentation. The acetate/ethanol ratio decreased as the pH declined, reaching close to 1 at pH 6.4. Whatever the pH conditions, lactate dehydrogenase was always greatly in excess. As pH decreased, both the biosynthesis and the catabolic efficiency of the pyruvate-ferredoxin oxidoreductase declined, as indicated by the ratio of the specific enzyme activity to the specific metabolic rate, which fell from 9.8 to 1.8. Thus a change of only 1 pH unit induced considerable metabolic change and ended by washout at around pH 6.2. C. cellulolyticum appeared to be similar to rumen cellulolytic bacteria in its sensitivity to acidic conditions. Apparently, the cellulolytic anaerobes studied thus far do not thrive when the pH drops below 6.0, suggesting that they evolved in environments where acid tolerance was not required for successful competition with other microbes. PMID- 11390678 TI - An NMR and enzyme study of the carbon metabolism of Neisseria meningitidis. AB - The pathogenic neisseriae are fastidious bacteria that are only able to grow on a restricted range of carbon sources. The genome sequence of Neisseria meningitidis strain MC58 predicts the presence of a complete citric acid cycle (CAC), but there have been no detailed biochemical studies of carbon metabolism in this important pathogen. In this study, both NMR and conventional enzyme assays were used to investigate the central metabolic pathways of a serogroup B strain (K454). (13)C-NMR labelling patterns of amino acids from hydrolysed cell proteins after growth with either 2- or 3-[(13)C]pyruvate were consistent with the operation of a complete oxidative CAC. Enzyme assays showed that cell-free extracts contained all the CAC enzymes predicted from the genome sequence, including a membrane-bound malate:quinone oxidoreductase which is present in place of the conventional NAD-linked cytoplasmic malate dehydrogenase. (1)H-NMR studies showed that growth on glucose, lactate and, especially, pyruvate, resulted in the excretion of significant amounts of acetate into the culture supernatant. This occurred via the phosphotransacetylase (PTA)-acetate kinase (ACK) pathway. Extremely high specific activities of PTA (7-14 micromol min(-1) mg(-1)) were detected in cell-free extracts, although ACK activities were much lower (46-298 nmol min(-1) mg(-1)). Expression of PTA and ACK activities was not co-ordinately regulated during growth on combinations of carbon sources. This may be related to the presence of two ackA paralogues in N. meningitidis which are, unusually, unlinked to the pta gene. PMID- 11390679 TI - Pyruvate oxidase contributes to the aerobic growth efficiency of Escherichia coli. AB - The metabolic importance of pyruvate oxidase (PoxB), which converts pyruvate directly to acetate and CO(2), was assessed using an isogenic set of genetically engineered strains of Escherichia coli. In a strain lacking the pyruvate dehydrogenase complex (PDHC), PoxB supported acetate-independent aerobic growth when the poxB gene was expressed constitutively or from the IPTG-inducible tac promoter. Using aerobic glucose-limited chemostat cultures of PDH-null strains, it was found that steady-states could be maintained at a low dilution rate (0.05 h(-1)) when PoxB is expressed from its natural promoter, but not at higher dilution rates (up to at least 0.25 h(-1)) unless expressed constitutively or from the tac promoter. The poor complementation of PDH-deficient strains by poxB plasmids was attributed to several factors including the stationary-phase dependent regulation of the natural poxB promoter and deleterious effects of the multicopy plasmids. As a consequence of replacing the PDH complex by PoxB, the growth rate (mu(max)), growth yield (Y(max)) and the carbon conversion efficiency (flux to biomass) were lowered by 33%, 9-25% and 29-39% (respectively), indicating that more carbon has to be oxidized to CO(2) for energy generation. Extra energy is needed to convert PoxB-derived acetate to acetyl-CoA for further metabolism and enzyme analysis indicated that acetyl-CoA synthetase is induced for this purpose. In similar experiments with a PoxB-null strain it was shown that PoxB normally makes a significant contribution to the aerobic growth efficiency of E. coli. In glucose minimal medium, the respective growth rates (mu(max)), growth yields (Y(max)) and carbon conversion efficiencies were 16%, 14% and 24% lower than the parental values, and correspondingly more carbon was fluxed to CO(2) for energy generation. It was concluded that PoxB is used preferentially at low growth rates and that E. coli benefits from being able to convert pyruvate to acetyl-CoA by a seemingly wasteful route via acetate. PMID- 11390680 TI - A peptidorhamnomannan from the mycelium of Pseudallescheria boydii is a potential diagnostic antigen of this emerging human pathogen. AB - The ascomycete Pseudallescheria boydii is an emerging human pathogen frequently found in soil and polluted water. A peptidopolysaccharide antigen has been isolated from mycelial forms of P. boydii, and characterized using chemical and immunological methods. Monosaccharide composition, methylation analysis, and (1)H and (13)C-NMR spectra indicated the presence of a rhamnomannan with a structure distinct from those of similar components isolated from other fungi, containing Rhap(1-->3)Rhap epitopes on side chains which may be linked (1-->3) to (1-->6) linked mannose. The peptidorhamnomannan from P. boydii reacted poorly with an antiserum raised against whole cells of Sporothrix schenckii and strongly with one against P. boydii hyphae. These characteristics and immunological differences suggest that this major rhamnose-containing antigen of P. boydii may be useful for the specific diagnosis of infections attributable to this fungus. PMID- 11390681 TI - A pre-genetic study of the isoforms of malic enzyme associated with lipid accumulation in Mucor circinelloides. AB - The oleaginous fungus Mucor circinelloides possesses at least six isoforms of malic enzyme (EC 1.1.1.40), a key lipogenic enzyme in filamentous fungi. These isoforms were detected using a specific stain for activity after native PAGE of cell extracts. Only one isoform (isoform IV) was associated with lipid accumulation, appearing only after N-exhaustion from the medium (which is a pre requisite for lipid accumulation) in glucose-growing cells. Isoforms I, II, V and VI were involved in anaerobic growth and only appeared under O(2)-limited conditions. Isoform III appeared to be constitutive and was formed under conditions of active (balanced) growth and is therefore thought to play a crucial role in basic metabolism. Growth on acetate increased the amount of cell lipid (from 25-27% in glucose-grown cells to 37-38% in acetate-grown cells) accumulated by M. circinelloides and this was associated with the appearance of isoform IV of malic enzyme prior to N-exhaustion in these cultures. Amino acid sequence analysis of isoforms III and IV suggests that these two malic enzymes may be encoded by a single gene and that isoform IV is formed from isoform III by post translational modification initiated by either N-limitation (when glucose was the carbon source) or growth on acetate as the sole carbon source. PMID- 11390682 TI - Involvement of a transformylase enzyme in siderophore synthesis in Pseudomonas aeruginosa. AB - Fluorescent pseudomonads produce yellow-green siderophores when grown under conditions of iron starvation. Here, the characterization of the pvdF gene, which is required for synthesis of the siderophore pyoverdine by Pseudomonas aeruginosa strain PAO1, is described. A P. aeruginosa pvdF mutant was constructed and found to be defective for production of pyoverdine, demonstrating the involvement of PvdF in pyoverdine synthesis. Transcription analysis showed that expression of pvdF was regulated by the amount of iron in the growth medium, consistent with its role in siderophore production. DNA sequencing showed that pvdF gives rise to a protein of 31 kDa that has similarity with glycinamide ribonucleotide transformylase (GART) enzymes involved in purine synthesis from a wide range of eukaryotic and prokaryotic species. Chemical analyses of extracts from wild-type and pvdF mutant bacteria indicated that the PvdF enzyme catalyses the formylation of N(5)-hydroxyornithine to give rise to N(5)-formyl-N(5)-hydroxyornithine, a component of pyoverdine. These studies enhance understanding of the enzymology of pyoverdine synthesis, and to the best of the authors' knowledge provide the first example of involvement of a GART-type enzyme in synthesis of a secondary metabolite. PMID- 11390683 TI - Substrate analysis and molecular cloning of the extracellular alkaline phosphatase of Streptomyces griseus. AB - Streptomyces species secrete large amounts of alkaline phosphatase (AP) enzymes that have not been characterized so far. An AP has been purified to homogeneity from cultures of Streptomyces griseus IMRU 3570. The enzyme has a monomer size of 62 kDa and is processed in the culture to a 33 kDa protein as shown by immunoblotting. The enzyme was purified by ammonium sulfate precipitation, CM Sephadex cationic exchange, chromatofocusing and HPLC Sphaerogel 3000SW filtration. The pure enzyme uses a variety of organic phosphorylated compounds as substrates. The N-terminal end of the mature protein was found to be RLREDPFTLGVASGDPHP. The gene phoA has been cloned using as probe an oligomer based on the N-terminal sequence of the S. griseus AP. phoA encodes a protein of 62678 Da with low homology to the AP of Escherichia coli. The phoA gene was found to be homologous to three alkaline-phosphatase-encoding genes previously identified in the Streptomyces coelicolor genome. On the basis of the optimal pH, substrate specificity and differences in amino acid sequence of motifs defining the active centre of APs, the S. griseus AP uses a wide range of organic phosphate substrates and is different from the phosphatases of Gram-negative bacteria. PMID- 11390684 TI - Cloning and functional analysis of a phosphopantetheinyl transferase superfamily gene associated with jadomycin biosynthesis in Streptomyces venezuelae ISP5230. AB - Sequence analysis of a XhoI/SacI fragment of chromosomal DNA downstream of jadL in the Streptomyces venezuelae ISP5230 gene cluster for jadomycin biosynthesis detected a partial ORF similar in its deduced amino acid sequence to the hetI product involved in synthesizing a regulator of heterocyst spacing in ANABAENA: By probing a phage library of S. venezuelae DNA with the XhoI/SacI fragment, the authors identified and isolated a hybridizing clone. The nucleotide sequence of its DNA contained three complete ORFs (jadM, N and X) and one incomplete ORF (jadO). The jadM ORF lay immediately downstream of, and partially overlapped, jadL. It contained 786 nucleotides encoding an amino acid sequence like those of enzymes in the phosphopantetheinyl transferase family. The jadN ORF contained 1794 nucleotides and encoded an amino acid sequence resembling acyl-CoA decarboxylases, thus suggesting a role in polyketide condensation reactions. The jadX ORF was not identified, but the partial jadO showed marked similarities in its deduced amino acid sequence to NDP-hexose-2,3-dehydratases, indicating a role in forming the sugar component of jadomycin B. Expression of jadM in Escherichia coli and examination of the product by SDS-PAGE established that the ORF encoded a 29.1 kDa protein, corresponding in size to the 262 amino acid polypeptide deduced from the jadM sequence. Evidence from a Northern hybridization indicated that jadM expression is correlated with jadomycin B synthesis. Cultures of S. venezuelae ISP5230 disrupted in jadM produced only 2-5% of the wild-type titre of jadomycin B, but grew well and produced chloramphenicol normally. The authors conclude that jadM encodes a holo-ACP synthase needed primarily for jadomycin B biosynthesis. PMID- 11390685 TI - Structure of the ask-asd operon and formation of aspartokinase subunits in the cephamycin producer 'Amycolatopsis lactamdurans'. AB - The first two genes of the lysine pathway are closely linked forming a transcriptional operon in the cephamycin producer 'Amycolatopsis lactamdurans'. The asd gene, encoding the enzyme aspartic semialdehyde dehydrogenase, has been cloned by complementation of Escherichia coli asd mutants. It encodes a protein of 355 aa with a deduced M(r) of 37109. The ask gene encoding the aspartokinase (Ask) is located upstream of the asd gene as shown by determination of Ask activity conferred to E. coli transformants. asd and ask are separated by 2 nt and are transcribed in a bicistronic 2.6 kb mRNA. As occurs in corynebacteria, the presence of a ribosome-binding site within the ask sequence suggests that this ORF encodes two overlapping proteins, Askalpha of 421 aa and M(r) 44108, and Askbeta of 172 aa and M(r) 18145. The formation of both subunits of Ask from a single gene (ask) was confirmed by using antibodies against the C-terminal end of Ask which is identical in both subunits. Ask activity of 'A. lactamdurans' is regulated by the concerted action of lysine plus threonine and this inhibition is abolished in E. coli transformants containing Ser(301) to Tyr, or Gly(345) to Asp mutations of the 'A. lactamdurans' ask gene. PMID- 11390686 TI - Unique expression of a highly conserved mycobacterial gene in IS901(+) Mycobacterium avium. AB - Expression of a gene encoding a novel protein antigen of 40 kDa (p40) was detected in IS901(+) strains of Mycobacterium avium, but not in any other species or subspecies of Mycobacterium tested, including IS901(-) M. avium and the other members of the M. avium complex. Although Southern hybridization revealed that the p40 gene is widely distributed within the genus, expression of the antigen could not be detected on Western blots of mycobacterial cell lysates. Nucleotide sequence analysis of the cloned p40 gene, and a database search, revealed high levels of sequence identity with a homologous gene in IS901(-) M. avium, M. avium subsp. paratuberculosis, Mycobacterium bovis, Mycobacterium leprae, Mycobacterium smegmatis and Mycobacterium tuberculosis. Further analysis of upstream sequences identified a putative promoter region. The p40 gene is the first example of a gene that is widely distributed within the genus Mycobacterium but expressed only in association with the presence of a genomic insertion element, in this case IS901, in strains of M. avium isolated from birds and domestic livestock. PMID- 11390687 TI - Genetic localization and regulation of the maltose phosphorylase gene, malP, in Lactococcus lactis. AB - Maltose phosphorylase (MP) from Lactococcus lactis was purified and the corresponding gene was cloned and expressed in Escherichia coli. The isoelectric point of the pure enzyme was determined to be 7.0. According to zymogram analysis and SDS-PAGE, the native MP was shown to be a monomeric enzyme with a molecular mass of 75 kDa. A polyclonal antiserum was produced to assess the regulation of the gene encoding MP in LC: lactis. According to immunoblot analysis, synthesis of the enzyme was markedly repressed by both glucose and lactose in the growth medium. When the lactococci were cultivated in the presence of other sugars, including maltose, trehalose or galactose, there was a pronounced expression of the MP gene. In addition, when the cells were grown in media without any added sugar, there was also pronounced expression of the enzyme, according to immunoblot analysis and specific activity data. These results indicated that no particular sugar specifically induces the gene encoding MP. However, an effect of glucose on MP expression was demonstrated by performing fermentations in the presence of both maltose and glucose. When glucose was added to maltose-grown lactococci in the mid-exponential growth phase, both the specific activity and amount of MP per millilitre of cell extract decreased rapidly. The genetic locus for the MP gene was found to be in the vicinity of the region encoding a possible regulator belonging to the LacI-GalR family of transcriptional regulators. Furthermore, this genetic location was separated from the previously characterized maltose-inducible and glucose-repressible beta-phosphoglucomutase (beta-PGM) gene. The different genetic loci for the genes encoding MP and beta PGM explains the different gene regulation behaviour. PMID- 11390688 TI - Analysis of sigma(54)-dependent genes in Enterococcus faecalis: a mannose PTS permease (EII(Man)) is involved in sensitivity to a bacteriocin, mesentericin Y105. AB - The sigma(54) RNA polymerase subunit has a prominent role in susceptibility of Listeria monocytogenes and Enterococcus faecalis to mesentericin Y105, a class IIa bacteriocin. Consequently, sigma(54)-dependent genes as well as specific activators also required for expression of these genes were sought. Five putative sigma(54)-associated activators were detected in the genome of E. faecalis V583, and all but one could activate the transcription of permease genes belonging to sugar phosphotransferase systems (PTSs). Interestingly, these activators display a helicase signature not yet reported in this activator family, which could explain the ATP-dependent mechanism of DNA unwinding preceding the start of transcription. To find which activator is linked to susceptibility of E. faecalis to mesentericin Y105, their respective genes were subsequently interrupted. Among them, only mptR gene interruption led to a resistance phenotype. Immediately downstream from mptR, a putative sigma(54)-dependent operon was found to encode a mannose PTS permease, namely EII(t)(Man). Moreover, in liquid culture, glucose and mannose induced the sensitivity of E. faecalis to mesentericin Y105. Since sugars have previously been reported to induce PTS permease expression, it appears that EII(t)(Man) expression, presumably induced in the presence of glucose and mannose, leads to an enhanced sensitivity of E. faecalis to the bacteriocin. Additional information was gained from knockouts within the permease operon. Interruption of the distal mptD gene, which encodes the IID subunit of EII(t)(Man), strikingly led to resistance to mesentericin Y105. Moreover, MptD appears to be a peculiar membrane subunit, bearing an additional domain compared to most known IID subunits. According to these results, EII(t)(Man) is clearly involved in susceptibility to mesentericin Y105 and could even be its receptor at the E. faecalis surface. Finally, it is hypothesized that MptD could be responsible for the targeting specificity, via an interaction between its additional domain and mesentericin Y105. PMID- 11390689 TI - Suppression of thermosensitive peptidyl-tRNA hydrolase mutation in Escherichia coli by gene duplication. AB - Peptidyl-tRNA hydrolase (Pth) in Escherichia coli is required to recycle tRNA molecules that dissociate from the ribosome as peptidyl-tRNA during protein synthesis. At non-permissive temperatures, strains with a thermosensitive mutation affecting the enzyme accumulate peptidyl-tRNA, cease protein synthesis and die. The rate of reversion of this mutation to thermoresistance varies widely according to the genetic background of the cell and the temperature of selection; under certain conditions, reversion can occur at rates approaching 10(-3) per cell per generation. In such revertants, a chromosomal pth gene can be replaced by an inactivated gene, restoring thermosensitive growth in most cases. PCR amplification experiments and Southern blots show the presence of both normal and inactivated copies of the gene, demonstrating that gene duplication has occurred in the revertants. Estimation of intracellular peptidyl-tRNA hydrolase by Western blotting confirms this explanation of the mechanism of high-frequency reversion to thermoresistance. PMID- 11390690 TI - Characterization of two autoreplicative regions of the IncHI2 plasmid R478: RepHI2A and RepHI1A((R478)). AB - Plasmids of the incompatibility groups HI and HII (IncH plasmids) generally confer multiple antibiotic resistances upon their host pathogenic strain. IncHI group plasmids are distinguished by their property of optimal transfer by conjugation at temperatures below 30 degrees C, allowing for the spread of multiple antibiotic resistance outside their host natural environment, the gut. Plasmids of the IncHI1 subgroup encode multiple replicons. In the present study it is shown that the prototype IncHI2 subgroup plasmid, R478, contains at least two iteron-controlled autoreplicative regions, RepHI2A and RepHI1A((R478)). The DNA sequence and the molecular characteristics of each replicon region are described. RepHI2A is unique to plasmids of the IncHI2 subgroup and contains an unusually large number of iteron sequences downstream of the replication initiator gene. The nucleotide sequence of the replication initiator gene and of the iterons within RepHI1A((R478)) show very close similarity with those of the previously reported RepHI1A replicon of the IncHI1 subgroup plasmid R27. The presence of RepHI1A((R478)) on R478 most likely accounts for the observed incompatibility between R478 and plasmids of the IncHI1 subgroup. These are the first autoreplicative regions from an IncHI2 subgroup plasmid to be described. PMID- 11390691 TI - Expression of Streptococcus mutans fimA is iron-responsive and regulated by a DtxR homologue. AB - Iron uptake, transport and storage in Streptococcus mutans, the principal causative agent of human dental cavities, is unexplored despite early reports in the literature which predict a role for this trace metal in cariogenesis. Experiments in the authors' laboratory revealed several iron-responsive proteins in S. mutans, one of which reacted with a polyclonal antiserum directed against the FimA fimbrial adhesin from Streptococcus parasanguis on Western blots. The results of Western blot and Northern hybridization experiments support an inverse relationship between iron availability and S. mutans fimA expression, and metal ion uptake experiments implicate FimA in S. mutans (55)Fe transport. Cloning of the S. mutans fimA homologue facilitated the construction of a fimA knockout mutant which grew poorly in an iron-limiting medium relative to the wild-type progenitor strain, lending further support to a role for FimA in S. mutans iron transport. The authors also identified and cloned a dtxR-like gene (dlg) located downstream of fimA on the S. mutans chromosome, and noted increased fimA expression in a S. mutans dlg knockout mutant relative to wild-type on RNA spot blots and Western blots. The uptake of (55)Fe, which was also significantly increased in this mutant, was compromised in a fimA/dlg double knockout. These findings are consistent with a role for Dlg in the iron-mediated regulation of fimA, and possibly other S. mutans iron transporters. Finally, the cariogenic potential of the fimA and dlg knockout mutants was not significantly different from that of the wild-type progenitor in a germ-free rat model. PMID- 11390692 TI - Regulation of the p-hydroxybenzoic acid hydroxylase gene (pobA) in plant-growth promoting Pseudomonas putida WCS358. AB - The regulation of the p-hydroxybenzoate hydroxylase gene (pobA) of Pseudomonas putida WCS358 involved in the catabolism of p-hydroxybenzoic acid (PHB) to the central intermediate protocatechuate was studied. Protocatechuic acid (PCA) is then degraded via the beta-ketoadipate pathway to form tricarboxylic acid intermediates. In several Gram-negative bacteria pobA has been found genetically linked to a regulator called pobR which activates pobA expression in response to PHB. In this study the identification and characterization of the pobC-pobA locus of P. putida WCS358 is presented. The p-hydroxybenzoate hydroxylase (PobA) is highly identical to other identified PobA proteins, whereas the regulatory protein PobC did not display very high identity to other PobR proteins studied and belonged to the AraC family of regulatory proteins, hence it has been designated POBC: Using the pobA promoter transcriptionally fused to a promoterless lacZ gene it was observed that induction via PobC occurred very efficiently when PHB was present and to a lesser but still significant level also in the presence of PCA. This PobC-PCA response was genetically demonstrated by making use of pobC::Tn5 and pcaH::Tn5 mutants of strain WCS358 constructed in this study. In pobC mutants both the p-hydroxybenzoic and PCA response were not observed, whereas in the pcaH mutant, which lacks a functional protocatechuate 3,4-dioxygenase, the protocatechuic-acid-dependent pobA activation was still observed. Finally, the activation of pobA by PHB varied according to the concentration and it was observed that in the pcaR::Tn5 regulatory mutant of strain WCS358 the pobA promoter activity was reduced. PcaR is a regulator involved in the regulation of several loci of the beta-ketoadipate pathway, one of which is pcaK. It was postulated that the reduction of pobA activation in pcaR::Tn5 mutants was because there was no expression of the pcaK gene encoding the PHB transport protein resulting in lower levels of PHB present inside the cell. PMID- 11390693 TI - Analysis of Pseudomonas putida alkane-degradation gene clusters and flanking insertion sequences: evolution and regulation of the alk genes. AB - The Pseudomonas putida GPo1 (commonly known as Pseudomonas oleovorans GPo1) alkBFGHJKL and alkST gene clusters, which encode proteins involved in the conversion of n-alkanes to fatty acids, are located end to end on the OCT plasmid, separated by 9.7 kb of DNA. This DNA segment encodes, amongst others, a methyl-accepting transducer protein (AlkN) that may be involved in chemotaxis to alkanes. In P. putida P1, the alkBFGHJKL and alkST gene clusters are flanked by almost identical copies of the insertion sequence ISPpu4, constituting a class 1 transposon. Other insertion sequences flank and interrupt the alk genes in both strains. Apart from the coding regions of the GPo1 and P1 alk genes (80-92% sequence identity), only the alkB and alkS promoter regions are conserved. Competition experiments suggest that highly conserved inverted repeats in the alkB and alkS promoter regions bind ALKS: PMID- 11390694 TI - Sulfur-limitation-regulated proteins in Bacillus subtilis: a two-dimensional gel electrophoresis study. AB - Little is known about the genes and enzymes involved in sulfur assimilation in Bacillus subtilis, or about the regulation of their expression or activity. To identify genes regulated by sulfur limitation, the authors used two- dimensional (2D) gel electrophoresis to compare the proteome of a wild-type strain grown with either sulfate or glutathione as sole sulfur source. A total of 15 proteins whose synthesis is modified under these two conditions were identified by matrix assisted laser desorption/ionization time of flight (MALDI TOF) mass spectrometry. In the presence of sulfate, an increased amount of proteins involved in the metabolism of C(1) units (SerA, GlyA, FolD) and in the biosynthesis of purines (PurQ, Xpt) and pyrimidines (Upp, PyrAA, PyrF) was observed. In the presence of glutathione, the synthesis of two uptake systems (DppE, SsuA), an oxygenase (SsuD), cysteine synthase (CysK) and two proteins of unknown function (YtmI, YurL) was increased. The changes in expression of the corresponding genes, in the presence of sulfate and glutathione, were monitored using slot-blot analyses and lacZ fusions. The ytmI gene is part of a locus of 12 genes which are co-regulated in response to sulfur availability. This putative operon is activated by a LysR-like regulator, YTLI: This is the first regulator involved in the control of expression in response to sulfur availability to be identified in B. subtilis. PMID- 11390695 TI - A novel glycosylated Cu/Zn-containing superoxide dismutase: production and potential therapeutic effect. AB - The fungal strain Humicola lutea 103 produces a naturally glycosylated Cu/Zn SOD. To improve its yield, the effect of an increased concentration of dissolved oxygen (DO) on growth and enzyme biosynthesis by the producer, cultivated in a 3 l bioreactor, was examined. Exposure to a 20% DO level caused a 1.7-fold increase of SOD activity compared to the DO-uncontrolled culture. Maximum enzyme productivity of SOD was approximately 300 x 10(3) U (kg wet biomass)(-1). The novel enzyme was purified to electrophoretic homogeneity. The presence of Cu and Zn were confirmed by atomic absorption spectrometry. The molecular mass of H. lutea Cu/Zn SOD was calculated to be 31870 Da for the whole molecule and 15936 Da for the structural subunits. The N-terminal sequence revealed a high degree of structural homology with Cu/Zn SOD from other prokaryotic and eukaryotic sources. H. lutea Cu/Zn SOD was used in an in vivo model for the demonstration of its protective effect against myeloid Graffi tumour in hamsters. Comparative studies revealed that the enzyme (i) elongated the latent time for tumour appearance, (ii) inhibited tumour growth in the early stage of tumour progression (73-75% at day 10) and (iii) increased the mean survival time of Graffi-tumour-bearing hamsters. Moreover, the fungal Cu/Zn SOD exhibited a strong protective effect on experimental influenza virus infection in mice. The survival rate increased markedly, the time of survival rose by 5.2 d and the protective index reached 86%. The H. lutea SOD protected mice from mortality more efficiently compared to the selective antiviral drug ribavirin and to commercial bovine SOD. In conclusion, our results suggest that appropriate use of the novel fungal SOD, applied as such or in combination with selective inhibitors, could outline a promising strategy for the treatment of myeloid Graffi tumour and influenza virus infection. PMID- 11390696 TI - A two-hybrid system based on chimeric operator recognition for studying protein homo/heterodimerization in Escherichia coli. AB - The development of a convenient and promising alternative to the various two hybrid methods that are used to study protein-protein interactions is described. In this system, a lambdoid chimeric operator is recognized by a hybrid repressor formed by two chimeric monomers whose C-terminal domains are composed of heterologous proteins (or protein domains). Only if these proteins efficiently dimerize in vivo is a functional repressor formed able to bind the chimeric operator and shut off the synthesis of a downstream reporter gene. This new approach was tested with several interacting proteins ranging in size from less than 100 to more than 800 amino acids and, to date, no size or topology limit has been detected. PMID- 11390697 TI - The osmotic stress response and virulence in pyelonephritis isolates of Escherichia coli: contributions of RpoS, ProP, ProU and other systems. AB - Trehalose synthesis (RpoS-dependent) and betaine uptake mediated by transporters ProP and ProU contribute to the osmotolerance of Escherichia coli K-12. Pyelonephritis isolates CFT073 and HU734 were similar and diminished in osmotolerance, respectively, compared to E. coli K-12. The roles of RpoS, ProP and ProU in osmoregulation and urovirulence were assessed for these isolates. Strain HU734 expressed an RpoS variant which had low activity and a C-terminal extension. This bacterium accumulated very little trehalose and had poor stationary-phase thermotolerance. For E. coli CFT073, introduction of an rpoS deletion impaired trehalose accumulation, osmotolerance and stationary-phase thermotolerance. The rpoS defects accounted for the difference in osmotolerance between these strains in minimal medium of very high osmolality (1.4 mol kg(-1)) but not in medium of lower osmolality (0.4 mol kg(-1)). The slow growth of both pyelonephritis isolates in high-osmolality medium was stimulated by glycine betaine (GB) and deletion of proP and/or proU impaired GB uptake. An HU734 derivative lacking both proP and proU retained osmoprotective GB uptake activity that could be attributed to system BetU, which is not present in strain K-12 or CFT073. BetU transported GB (K(m), 22 microM) and proline betaine. High osmolality human urine (0.92 mol kg(-1)) included membrane-permeant osmolyte urea (0.44 M) plus other constituents which contributed an osmolality of only approximately 0.4 mol kg(-1). Strains HU734 and CFT073 showed correspondingly low GB uptake activities after cultivation in this urine. Deletion of proP and proU slowed the growth of E. coli HU734 in this high-osmolality human urine (which contains betaines) but had little impact on its colonization of the murine urinary tract after transurethral inoculation. By contrast, deletion of rpoS, proP and proU had no effect on the very rapid growth of CFT073 in high-osmolality urine or on its experimental colonization of the murine urinary tract. RpoS dependent gene expression is not essential for growth in human urine or colonization of the murine urinary tract. Additional osmoregulatory systems, some not present in E. coli K-12 (e.g. BetU), may facilitate growth of pyelonephritis isolates in human urine and colonization of mammalian urinary tracts. The contributions of systems ProP and ProU to urinary tract colonization cannot be definitively assessed until all such systems are identified. PMID- 11390698 TI - Commensal Escherichia coli isolates are phylogenetically distributed among geographically distinct human populations. AB - An intraspecies phylogenetic grouping of 168 human commensal Escherichia coli strains isolated from the stools of three geographically distinct human populations (France, Croatia, Mali) was generated by triplex PCR. The distributions of seven known extraintestinal virulence determinants (ibeA, pap, sfa/foc, afa, hly, cnf1, aer) were also determined by PCR. The data from the three populations were compiled, which showed that strains from phylogenetic groups A (40%) and B1 (34%) were the most common, followed by phylogenetic group D strains (15%). Strains of the phylogenetic group B2 were rare (11%). However, a significant specific distribution for strains of groups A, B1 and B2 within each population was observed, which may indicate the influence of (i) geographic/climatic conditions, (ii) dietary factors and/or the use of antibiotics or (iii) host genetic factors on the commensal flora. Virulence determinants were rarely detected, with only 25.6% of the strains harbouring at least one of the virulence genes tested. The strains with virulence factors most frequently belonged to phylogenetic group B2. The commensal strains of phylogenetic groups A, B1 and D had fewer virulence determinants than pathogenic strains of the corresponding groups when these data were compared with those for previous collections of virulent extraintestinal infection strains studied using the same approach. However, the virulence patterns of commensal and pathogenic B2 phylogenetic group strains were the same. The data thus suggest that strains of the A, B1 and D phylogenetic groups predominate in the gut flora and that these strains must acquire virulence factors to become pathogenic. In contrast, commensal phylogenetic group B2 strains are rare but appear to be potentially virulent. PMID- 11390699 TI - The role of antibodies to Bacillus anthracis and anthrax toxin components in inhibiting the early stages of infection by anthrax spores. AB - Vaccines which are efficacious against anthrax, such as the human vaccine, Anthrax Vaccine Absorbed (AVA), contain the protective antigen (PA) component of the anthrax toxins as the major protective immunogen. Although AVA protects against inhalational anthrax, the immune responses to and role in protection of PA and possibly other antigens have yet to be fully elucidated. Sera from animals immunized with a toxin-producing, unencapsulated live vaccine strain of Bacillus anthracis have been reported to have anti-spore activities associated with the antitoxin humoral response. The authors performed studies to determine whether anti-PA antibody (Ab)-containing preparations stimulated spore uptake by phagocytes and suppressed the germination of spores in vitro. AVA- and PA-immune sera from several species enhanced the phagocytosis by murine peritoneal macrophages of spores of the virulent Ames and the Sterne vaccine strains. Antitoxin Abs appeared to contribute significantly, although not solely, to the enhanced uptake. Rabbit antisera to PA purified from either Sterne or a PA producing pX01-cured recombinant, affinity-purified anti-PA IgG, and monkey antisera to AVA were used to assess the role of anti-PA ABS: Rabbit anti-PA Abs promoted the uptake of spores of the PA-producing strains Sterne, Ames and RP42, a mutant of Sterne producing only PA, but not of the pX01-Sterne-1 strain, Ames strain, or RP4, a mutant of Sterne with deletions in the loci encoding PA and the oedema factor (EF) toxin component and producing only the lethal factor toxin component. Rabbit anti-PA and monkey anti-AVA Abs also significantly inhibited spore germination in vitro compared to preimmune serum or medium. Spore associated proteins recognized by anti-PA Abs were detected by electron microscopy and confirmed by immunoblotting of spore coat extracts. Thus, the anti PA Ab-specific immunity induced by AVA has anti-spore activity and might have a role in impeding the early stages of infection with B. anthracis spores. PMID- 11390700 TI - Evidence for a more recently evolved clade within a Candida albicans North American population. AB - Candida albicans is diploid and displays a primarily clonal mode of reproduction. There is, however, evidence for meiosis and the degree to which this occurs in nature is unknown. Although random mating would act to obscure clonal lineages, previous studies have demonstrated that collections of North American isolates display three major partitions with no evidence of geographic clustering. To better understand the extent of sexuality and its role in the phylogeny of the species, a reference subset of 50 isolates representing this tripartite division was analysed using 1 minisatellite, 5 microsatellites (MSs) and 15 nuclear polymorphisms (NP). A total of 87 alleles were observed for 21 loci and 12/16 informative loci exhibited a departure from Hardy-Weinberg expectations (G(2)50% luminal diameter in 13 patients. Quantitative angiographic evaluation was performed in a total of 45 segments showing >25% narrowing at the second angiogram. A progression (>15% luminal reduction) was found in 17 of 45 segments, a new lesion (initial luminal diameter <20%) was detected in nine segments, resulting in progression or new lesion in 16 patients (62%). Patients with or without progression did not differ in age, duration of dialysis treatment, number of cardiovascular risk factors, or serum total cholesterol and fibrinogen levels. After percutaneous transluminal coronary angioplasty (PTCA) a restenosis was seen in seven of 16 primarily successfully dilated segments. After the second angiography, myocardial revascularization was performed in eight patients (1 PTCA, 7 coronary artery bypass graft). CONCLUSIONS: Patients with end stage renal disease have a high prevalence of CAD. In line with the clinical course, CAD patients on maintenance dialysis undergo rapid angiographic progression of CAD, which results in a high rate of subsequent myocardial revascularizations. PMID- 11390721 TI - Inferior vena cava diameter: a useful method for estimation of fluid status in children on haemodialysis. AB - BACKGROUND: An accurate assessment of fluid status in haemodialysis patients presents a significant challenge especially in growing children. Clinical parameters of hydration are not always reliable, and invasive methods such as measurement of central venous pressure cannot be used routinely. We evaluated the usefulness of inferior vena cava diameter (IVCD) measured by echocardiography in the estimation of hydration in children on haemodialysis. METHODS: Fifteen haemodialysis patients (mean age 14 years) were evaluated. Clinical assessment included patients' symptoms, weight, blood pressure, heart rate, presence of oedema and vascular congestion, before and after dialysis session. Dry weight was assessed based on the above parameters. Fifty-two echocardiographic studies immediately prior and 30-60 min following dialysis were performed. The anteroposterior IVCD was measured 1.5 cm below the diaphragm in the hepatic segment in supine position during normal inspiration and expiration. IVCD was expressed per body surface area. RESULTS: Following haemodialysis mean IVCD (average of expiration and inspiration) decreased from 1.12+/-0.38 to 0.75+/-0.26 cm/m(2) (P<0.0001). Changes in IVCD were significantly correlated with alterations in body weight following dialysis (P<0.0001). The collapse index (per cent of change in IVCD in expiration vs inspiration) increased significantly after dialysis (P=0.035). IVCD clearly reflected alterations in fluid status. It did not vary significantly with changes in dry weight in a given patient. CONCLUSIONS: Our findings support the applicability of VCD measurement in the estimation of hydration status in paediatric haemodialysis patients. The combination of clinical parameters and measurement of IVCD may enable more accurate evaluation of hydration of children on haemodialysis. PMID- 11390722 TI - Predilution haemofiltration--the Second Sardinian Multicentre Study: comparisons between haemofiltration and haemodialysis during identical Kt/V and session times in a long-term cross-over study. AB - BACKGROUND: The potential superiority of various renal replacement treatment modalities consisting largely of convective mass transfer as opposed to primarily diffusive mass transfer, is still a matter of debate. The objective of the present study was to evaluate acute and long-term clinical effects of varying degrees of convection and diffusion in a group of 24 clinically stable patients with end-stage renal disease. METHODS: The patients were prospectively assigned to three consecutive treatment schedules of 6 months each: phase I (HF1) (on-line predilution haemofiltration)-->phase II (HD) (high-flux haemodialysis)-->phase III (HF2; as phase I). We used the AK100/200 ULTRA monitor (Gambro), which prepares ultrapure dialysis fluid for HD and sterile, pyrogen-free substitution solution for HF. The membrane (polyamide), fluid composition, and treatment time were the same on HF and HD. The targeted equilibrated Kt/V was 1.2 for both treatment modes, creating a similar urea clearance. RESULTS: Fifteen patients, mean age 62.8+/-8.4 years, completed the study according to the above conditions. Urea kinetics, nutritional parameters, and dry weight were similar in the three periods. The frequency of intra-treatment episodes of hypotension/patient/month was significantly lower on HF1 (1.24) and HF2 (1.27) than on HD (1.80) (P<0.04). It decreased progressively on HF1, then increased on HD, and decreased again during HF2. Patients had fewer muscular cramps on HF than on HD (P<0.03) and required significantly less saline and plasma expander during HF than HD sessions. The prevalence of inter-treatment symptoms, including fatigue and hypotension, was lower on HF than on HD (score difference P=0.04). Quality of life, determined by the Laupacis method in all three periods, showed a tendency towards improvement during the study, reaching the best values during HF2. CONCLUSIONS: HF has a progressive stabilizing haemodynamic effect, producing a more physiological cardiovascular profile than HD. This long-term effect, observed in stable patients treated under strictly identical conditions, is probably due to the mechanism of convection, and is different from the acute effect observed mainly in unstable patients. PMID- 11390723 TI - Rate dependence of acute PTH release and association between basal plasma calcium and set point of calcium-PTH curve in dialysis patients. AB - BACKGROUND: In vivo, the control of calcium-mediated acute PTH release during induced hypo- or hypercalcaemia is linked not only to plasma calcium concentration per se but also to the rate and direction of calcium change. In fact, during induced hypocalcaemia, the predominant mechanism that causes PTH to be released is the reduction of plasma Ca(2+) irrespective of the absolute starting concentration of ionized calcium. This mechanism, which is rate dependent and even activated in conditions of hypercalcaemia, may be involved in the association, reported in several papers, between the basal Ca(2+) and the set point of the calcium-PTH curve. METHODS: The calcium-PTH relationship was studied in 12 dialysis patients under conditions of induced low and high predialysis plasma Ca(2+). At each level of basal Ca(2+), dynamic tests were conducted using two methodological approaches. In method A patients underwent low (0.5 mmol/l) calcium dialysis in the stimulation test and high (2 mmol/l) calcium dialysis in the inhibition test, while the dialysate calcium (CaD) was kept constant during each test. In this way a higher but variable rate of change in plasma Ca(2+) was achieved. In method B, CaD was progressively decreased (stimulation test) and increased (inhibition test) during the tests in order to obtain a lower but more constant rate of change in plasma Ca(2+). Consequently, for each patient, four calcium-PTH curves were produced: low basal Ca(2+) with methods A and B, and high basal Ca(2+) with methods A and B. RESULTS: Basal plasma Ca(2+) was similar in A and B at low (1.16+/-0.02 vs 1.15+/-0.02 mmol/l) and high (1.25+/-0.02 vs 1.26+/ 0.02 mmol/l) basal plasma Ca(2+). The set point was higher in A than in B both at low (1.12+/-0.02 vs 1.10+/-0.02 mmol/l, P=0.01) and high (1.20+/-0.02 vs 1.16+/ 0.02 mmol/l, P=0.03) basal Ca(2+) as was the slope (542+/-41 vs 426+/-44%/mmol, P=0.02; 615+/-73 vs 389+/-25%/mmol, P=0.01). No significant difference was found between A and B as regards minimal PTH and plasma Ca(2+) at minimal PTH (Camin) in both calcaemic states. Maximal PTH was slightly higher in B at low (510+/-97 vs 548+/-107 pg/ml, P=NS) and high basal plasma Ca(2+) (410+/-97 vs 464+/-108 pg/ml, P=0.02). Plasma calcium at maximal PTH (Camax) was significantly higher in A (1.1+/-0.03 vs 0.99+/-0.02 mmol/l, P=0.001) at high basal plasma Ca(2+). The set point was strictly related to basal plasma Ca(2+) in both methods, but the slope of the linear regression was significantly steeper with method A. The set point was predicted to increase by 0.881 (CI 0.772-0.990) mmol/l for each mmol/l of increase in basal plasma Ca(2+) with method A and by 0.641 (CI 0.546-0.737) mmol/l for each mmol/l of increase in basal plasma Ca(2+) with method B. CONCLUSIONS: (i) Higher and variable rates of change in plasma Ca(2+) produce a higher set point value and a steeper slope of the calcium-PTH curve when compared to lower and more constant rates of calcium change. (ii) The different slope of the linear correlations between basal plasma Ca(2+) and set point in the two methods suggests that the rate-dependent mechanism of acute PTH release plays a significant role in the association between set point and basal plasma Ca(2+). (iii) The significance of the set point is questionable when the calcium-PTH curve is carried out in vivo. PMID- 11390724 TI - Intra- and post-dialytic platelet activation and PDGF-AB release: cellulose diacetate vs polysulfone membranes. AB - BACKGROUND: During haemodialysis the blood-membrane contact causes a release of platelet granule content, which contains platelet-derived growth factor AB (PDGF AB). In view of the potential role of this in altering biocompatibility during haemodialysis, we evaluated the intra- and post-dialytic changes in PDGF-AB serum levels during haemodialysis sessions performed with cellulose diacetate (CDA) and polysulfone (PS) membranes respectively. METHODS: PDGF-AB, platelet factor 4 (PF4), beta thromboglobulin (betaTG), and mean platelet volume (MPV) levels were determined in 30 patients, each of whom underwent six dialysis sessions: three with a CDA and three with a PS membrane. Blood samples were taken at times 0, 15, 30, 120, 180, and 240 min during dialysis and at 1, 4, and 20 h after the end of the session. Statistical analysis was performed using a one-way ANOVA and Student's t test. RESULTS: PDGF-AB at 15 min was increased to +41+/-9% with CDA vs +20+/-5% with PS (P<0.001) from the T0 values, and at 120 min it was +19+/-8% with CDA vs -25+/-9% with PS (P<0.001) from T0 levels. At 240 min it was +95+/ 14% with CDA vs +49+/-15% with PS (P<0.001) from the T0 values, returning to basal only 20 h after the end of the session. betaTG at 15 min was +60+/-8% for CDA vs +24+/-7.5% for PS (P<0.001) from the T0 values. PF4 showed a similar trend to betaTG. MPV at 30 min from the start of dialysis was 7.4+/-0.3 fl with CDA and 8+/-0.3 fl with PS (P<0.001), and at 240 min MPV was 7.9+/-0.3 fl with CDA and 8.4+/-0.3 fl with PS (P<0.001). CONCLUSIONS: Platelet activation and platelet release reactions are lower with PS than with CDA membranes. PDGF-AB, released during and after dialysis, represents a clear biocompatibility marker. Its slow return to basal values and its action on vascular cells make it a potential risk factor for atherosclerosis in uraemic patients. PMID- 11390725 TI - Effect of type of dialysis membrane on bone in haemodialysis patients. AB - BACKGROUND: Uraemic bone disease is the result of a number of factors modulating bone formation and resorption in a complex manner. In the present study, the hypothesis tested was that the type of haemodialysis membrane used for renal replacement therapy might also play a role. METHODS: We conducted a prospective, open study in 24 chronic haemodialysis patients who were randomized to dialysis treatment with either cellulosic (CELL group, n=11) or polyacrylonitrile (AN-69 group, n=13) membrane for 9 months. Repeated determinations of plasma parameters reflecting bone turnover were done in all patients, and a bone biopsy in a subgroup at the start and end of study. RESULTS: At the start, mean plasma intact parathyroid hormone levels were comparable between the two groups and they did not vary significantly at 9 months of treatment. Similarly, plasma bone-specific alkaline phosphatase and osteocalcin (markers of bone formation), and cross-laps (marker of bone resorption) remained unchanged. However, plasma insulin-like growth factor-I (IGF-I) progressively decreased from 169 to 119 ng/ml in AN-69 group (P<0.01), whereas it remained unchanged in CELL group. In addition, the levels of IGF binding protein (IGFBP)-1 and IGFBP-2 were increased while the levels of IGFBP-5 were decreased in AN-69 group. In the five patients of each group who had repeat bone biopsies, histomorphometric analysis showed a decrease in osteoblast surface, osteoclast surface and osteoclast number in AN-69 group at 9 months, compared with baseline values measured at the start of the study. In contrast, all three parameters significantly increased in the CELL group at 9 months (P<0.001 for the difference between each of the three parameters). Bone formation rate decreased by 31% in the AN-69 group, but increased by 50% in CELL group. However, this latter difference was not statistically significant. Plasma interleukin (IL)-6 and soluble IL-6 receptor levels did not change in the two groups of patients who had undergone bone biopsy. CONCLUSION: Dialysis with CELL membrane was associated with increased bone turnover whereas the use of AN-69 membrane was associated with decreased bone turnover, suggesting a beneficial effect of the latter on high-turnover uraemic bone disease. However, as the number of patients with repeat bone biopsies was small, these findings need to be confirmed in a larger study. Further studies are also needed to evaluate whether or not the changes in IGF system components play a role in decreased bone cell activity in patients on dialysis using the AN-69 polyacrylonitrile membrane. PMID- 11390726 TI - A randomized, controlled parallel-group trial on efficacy and safety of iron sucrose (Venofer) vs iron gluconate (Ferrlecit) in haemodialysis patients treated with rHuEpo. AB - BACKGROUND: The objectives of the present trial were to compare the efficacy and safety of two i.v. iron preparations with respect to haemoglobin levels, iron status and recombinant human erythropoetin (rHuEpo) dosage requirements in stable, rHuEpo-treated haemodialysis patients (maintenance phase of iron treatment) over 6 months. METHODS: A total of 59 patients were randomized and assigned to one of two treatment groups and 55 patients were analysed (iron sucrose n=27; iron gluconate n=28). Iron sucrose was administered in a dose of 250 mg iron diluted in 100 ml normal saline given over 60 min once per month, while 62.5 mg iron as iron gluconate was given once per week in a slow push injection (5 min). RESULTS: --Efficacy parameters: Haemoglobin levels could be maintained from baseline to endpoint in both groups. There were, however, more patients in the iron sucrose group than in the iron gluconate group for whom treatment was discontinued because their haemoglobin values exceeded 12.5 g/dl or ferritin values exceeded 1000 ng/ml (five vs two and three vs one patient, respectively). Transferrin saturation and serum ferritin increased significantly in both groups (+255.7 ng/ml with iron sucrose and +278.5 ng/ml with iron gluconate), while rHuEpo dosage did not change significantly throughout the study. --Safety parameters: There were a total of 174 infusions of iron sucrose and 720 injections of iron gluconate during the trial; all of them were well tolerated. In particular, we did not observe anaphylactoid reactions or any events suggestive of iron toxicity such as hypotension, dizziness, or nausea. CONCLUSIONS: High doses of iron sucrose (Venofer((R)) at a dose of 250 mg/month) was equally effective in maintaining haemoglobin and equally well tolerated as low doses of iron gluconate (Ferrlecit((R)) at a dose of 62.5 mg once per week) in stable, rHuEpo treated haemodialysis patients. PMID- 11390727 TI - Association of sex hormone status with the bone loss of renal transplant patients. AB - BACKGROUND: Bone loss is an important problem in renal transplantation recipients. The role of sex hormones in this setting has not been previously addressed. The objective was to investigate whether sex hormone status is associated with bone mass loss in renal transplant recipients. METHODS: Thirty patients (16 men and 14 women, of which eight were post-menopausal) were studied by bone densitometry and bone biopsy. In women, serum oestradiol levels and menopausal status were determined; in men, serum testosterone levels were assessed. RESULTS: Mean age was 48+/-11 years. Time on dialysis was 13+/-17 months, and time since transplantation was 125+/-67 months. Thirteen patients were on cyclosporine A (CsA) monotherapy, 12 on azathioprine plus prednisolone (PRED) dual therapy, and five on CsA, azathioprine and PRED triple therapy. In men, serum testosterone levels were 19.7+/-6.8 nmol/l (mean+/-SD). In pre menopausal women, oestradiol serum levels were 209(128-289) pmol/l (median (percentiles 25-75%)), and in post-menopausal women 93(54-299) pmol/l (non significant). Univariate analysis in women demonstrated that serum oestradiol levels were positively correlated with Z scores of osteoblast surface (r=0.70, P=0.005), osteoid surface (r=0.75, P=0.002) and trabecular wall thickness (r=0.68, P=0.008). In men, a weak correlation was seen between serum testosterone levels and the cumulative dose of PRED (r=-0.52, P=0.06). In the multivariate analysis, two models of multiple regression were employed (one for women and one for men), considering the densitometric and histomorphometric variables (Z scores) as dependent variables. Serum testosterone in men did not predict any of the densitometric nor histomorphometric variables analysed, while serum oestradiol in women was an independent predictor for the osteoblast surface (r=0.81, P=0.003), osteoid surface (r=0.82, P=0.009) and trabecular wall thickness (r=0.54, P=0.05). CONCLUSIONS: In female renal transplant recipients, serum oestradiol levels independently predict the bone status, while in men, factors other than testosterone seem to influence bone loss. Our results give rise to the hypothesis that sex hormone replacement therapy may play a role in prevention and/or treatment of the bone loss in women following renal transplantation. PMID- 11390729 TI - Microemulsion cyclosporin formulation, in contrast to the old formulation, widens the T lymphocyte subsets differences between stable and acute rejection of kidney transplants. AB - BACKGROUND: The new cyclosporin (CsA) formulation, Neoral, has different pharmacokinetics compared with Sandimmune (SIM). Larger area under the curve (AUC) values with equivalent trough blood values are reached when Neoral is administered at equivalent doses to SIM. Previously, we showed a great diagnostic reliability when using cytofluorometric analysis from fine-needle aspiration biopsy (FNAB) samples. We investigated possible changes brought about by Neoral on lymphocyte subsets and the repercussions on the activation score cut-off for acute rejection, defined under SIM treatment. METHODS: Of 63 patients that received SIM, 40 remained rejection-free and 23 suffered one episode of rejection. Of 52 patients that received Neoral, 38 remained rejection-free. Peripheral blood lymphocytes (PBL) and lymphocytes from FNAB taken on days 7 and 14 post-transplantation and on the first day of acute rejection were analysed by flow cytometry. RESULTS: Trough blood CsA levels were not different between SIM and Neoral treatments. Among rejection-free patients, a significant down regulation of CD3DR and of CD8DR expression on both graft-infiltrating lymphocytes (GIL) and PBL, and significant up-regulation of naive T cells on GIL were observed with Neoral. These changes were followed by a significant down regulation of the activation score with Neoral. Conversely, within the acute rejection group, the activation score was significantly higher with Neoral than with SIM. The activation score performed equally well in Neoral transplants compared with what we had reported with SIM. CONCLUSIONS: Our study indicates that Neoral elicits stronger immunosuppressive effects in stable patients, which eventually should translate into better clinical efficiency. However, when acute rejection supervenes, the treatment breakthrough seems stronger with Neoral. Cytofluorometric studies from FNAB samples showed that diagnostic reliability was maintained at a high level under Neoral therapy. PMID- 11390728 TI - Vitamin C improves endothelial dysfunction in renal allograft recipients. AB - BACKGROUND: Endothelial function is impaired in renal allograft recipients but the effects of antioxidant vitamin therapy on endothelial function in such patients is unknown. METHODS: Thirteen renal allograft recipients were randomized to vitamin C or placebo in a double blind cross-over study design. Flow-mediated endothelium-dependent dilation and glyceryltrinitrate-induced endothelium independent dilation of the brachial artery were assessed before and 2 h after oral administration of 2 g vitamin C or placebo. RESULTS: Plasma vitamin C levels increased from 33.5+/-17.0 micromol/l to 98.8+/-60.2 micromol/l after treatment (P=0.0001). Endothelium-dependent dilation improved (from 1.6+/-2.6 to 4.5+/ 2.5%) after vitamin C administration but was unchanged after placebo (1.9+/-1.5 to 1.8+/-2.5%; P=0.003 for vitamin C vs placebo). There was no significant change in endothelium-independent dilation in response to vitamin C. Vitamin C was also associated with a significant increase in the lag time in dilute serum oxidation (P=0.001). CONCLUSIONS: Vitamin C acutely improves flow-mediated, endothelium dependent dilation and increases the resistance of lipoproteins in dilute serum to oxidation in renal transplant recipients. PMID- 11390730 TI - An experimental sclerosing encapsulating peritonitis model in mice. AB - BACKGROUND: Sclerosing encapsulating peritonitis (SEP) is a life-threatening complication of continuous ambulatory peritoneal dialysis. To elucidate the mechanism and develop treatments for this condition, an experimental SEP model in mice was constructed. METHODS: C57BL/6 mice were administered 0.3 ml of 0.1% chlorhexidine gluconate and 15% ethanol dissolved in saline, intraperitoneally, on a daily basis for 56 days (group 1, n=15). A control group of C57BL/6 mice were administered 0.3 ml of phosphate-buffered saline only in the same manner (group 2, n=15). The mice were sacrificed on days 3, 7, 21, 56 and were prepared for histological analysis. RESULTS: In group 1, all mice had developed macroscopic evidence of SEP 56 days after the injection. Microscopically we observed peritoneal fibrosis and an increase in infiltrates of mononuclear cells over time. The peritoneal fibrosis reached the chronic inflammatory stage by 56 days after the injection. CONCLUSION: We have developed a convenient experimental model of SEP in mice, which may be useful in elucidating the pathogenesis of SEP and in establishing possible treatments. PMID- 11390731 TI - Kidney disease in hypomelanosis of Ito. PMID- 11390732 TI - Tryptophan immunoadsorption strongly reduces proteinuria in recurrent nephrotic syndrome. PMID- 11390733 TI - Severe obesity in haemodialysis: the utility of bioimpedance vector analysis. PMID- 11390734 TI - Type 1 glycogen storage disease and recurrent calcium nephrolithiasis. PMID- 11390735 TI - Possible mechanisms to explain the absence of hyperkalaemia in Addison's disease. PMID- 11390736 TI - A rare infection in a renal transplant recipient. PMID- 11390737 TI - On the history of the study of haemorrhagic fever with renal syndrome in eastern Russia. PMID- 11390739 TI - Renal diagnosis without renal biopsy. Nephritis and sensorineural deafness. AB - Two examples of hereditary nephropathy within the context of clinical syndromes are described. Emphasis is put on the ability to make a renal diagnosis without renal biopsy and the benefits of screening relatives once a diagnosis is achieved. A variant of Alport's syndrome with associated macrothrombocytic thrombocytopenia, known as Epstein's syndrome, is reported. In addition siblings with Alstrom's syndrome characterized by pigmentary retinal degeneration (causing blindness in early childhood), progressive sensorineural hearing loss, and progressive renal failure are reported. Both cases had previously presented for non-renal pathology in advance of the onset of symptomatic renal failure and may have benefited from appropriate screening. PMID- 11390740 TI - John Walls (1939-2001): an appreciation. PMID- 11390742 TI - The advanced fetal programming hypothesis. PMID- 11390743 TI - Minimal-change nephrotic syndrome associated with mixed connective-tissue disease. PMID- 11390744 TI - Relationship of renal dysfunction to proximal arterial disease severity in atherosclerotic renovascular disease. PMID- 11390745 TI - Effectiveness of haemodialysis with high-flux membranes in the extracorporeal therapy of life-threatening acute lithium intoxication. PMID- 11390746 TI - Fibrate treatment can increase serum creatinine levels. PMID- 11390747 TI - Use of urea containing dialysate to avoid disequilibrium syndrome, enabling intensive dialysis treatment of a diabetic patient with renal failure and severe metformin induced lactic acidosis. PMID- 11390748 TI - Corticosteroid therapy in encapsulating peritoneal sclerosis. PMID- 11390750 TI - How We Smell: The Molecular and Cellular Bases of Olfaction. AB - Three models for the perception of odor molecules are suggested for the first time by experimental data. These studies illustrate how the nose may smell. Moreover, they suggest additional role(s) for odor receptors within and outside the olfactory system. PMID- 11390751 TI - Renal Proximal Tubular Albumin Reabsorption: Daily Prevention of Albuminuria. AB - Although the glomerular filtration coefficient of albumin is small, the daily filtered load can be as much as 8 g. To prevent such massive losses of albumin, quantitative reabsorption along the proximal tubules is accomplished by "receptor"-mediated endocytosis. Albumin reaches the lysosomes where it is degraded to amino acids. PMID- 11390752 TI - Structure and Function of Renal Organic Cation Transporters. AB - Polyspecific transport systems in the kidney mediate the excretion and reabsorption of organic cations. Electrogenic import systems and electroneutral export systems in the basolateral and luminal plasma membranes of proximal renal tubules are involved. Two subtypes of electrogenic import systems have been cloned from rats and humans and functionally characterized. PMID- 11390753 TI - Killing Mechanisms of Cytotoxic T Lymphocytes. AB - Cytotoxic T lymphocytes mediate lysis of target cells by various mechanisms, including exocytosis of lytic proteins (perforin, granzymes) and receptor-ligand binding of Fas/APO molecules. Death of target cells is characterized by either necrosis or apoptosis, depending on the killing mechanism used and on the metabolism of the target cell itself. PMID- 11390754 TI - Antistress Pattern Induced by Oxytocin. AB - Repeated oxytocin injections cause lowered blood pressure, decreased cortisol levels, increased withdrawal latency, increased release of vagally controlled gastrointestinal hormones, and increased weight gain. Together, these effects form an antistress pattern. Nonnoxious sensory stimuli release oxytocin and induce an effect spectrum similar to the one caused by oxytocin injections. PMID- 11390755 TI - Organizational Principles of Cerebellar Neuronal Circuitry. AB - We review our recent studies of cerebellar neuronal organization, emphasizing that consideration of organizational features of cerebellar circuitry represents a necessary step toward the understanding of how the cerebellum does what it does, in terms of both its internal information processing and its interaction with other motor structures. PMID- 11390756 TI - The Tuned Cricket. AB - Communication by means of sound is not always easy. Sound suffers much attenuation and degradation close to ground. Crickets have adapted to this by exploiting sharply tuned mechanical systems. A coevolution ensures that the calling song of males and the directional hearing of females are tuned to the same frequency. PMID- 11390757 TI - Understanding the Role of Paracellular Transport in the Proximal Tubule. AB - Fluid and solute reabsorption by the proximal tubule is the result of both transcellular and paracellular flux. The role of transcellular transport has been extensively studied, but the importance of paracellular flux has not been as thoroughly investigated. The purpose of this review is to update concepts about the contribution of paracellular transport for reabsorption by the proximal tubule. PMID- 11390759 TI - In Forthcoming Issue. PMID- 11390758 TI - Ion Permeability of the Nuclear Envelope. AB - The nuclear envelope mediates nucleocytoplasmic communication. Nuclear pores transport proteins and RNA into and out of the nucleus. The pore is believed to allow free ion diffusion. Using an electrophysiological approach, we show the possible semipermeable properties of the envelope. To accomplish these functions we hypothesize a mechanism in which the pore complex acts as a molecular diaphragm. PMID- 11390760 TI - Cyclic Nucleotide Phosphodiesterases: New Role in the Pathogenesis of Glomerulonephritis? PMID- 11390762 TI - In Forthcoming Issue. PMID- 11390761 TI - From Genetically Altered Mice to Integrative Physiology. AB - Transgenic and gene-targeted mice permit the study of the function(s) of the single gene(s) in a whole organism, thereby relating molecular biology and integrative physiology. To demonstrate the potential of transgenic models, we present in this review some physiologically relevant information obtained from genetically engineered mice. PMID- 11390763 TI - Nerves and Intestinal Mast Cells Modulate Responses to Enterotoxins. AB - Experiments in intact animals exposed to enterotoxins demonstrate that neurons and immune cells of the lamina propria regulate toxin-induced diarrhea and tissue damage. Clostridium difficile toxins cause profound diarrhea and acute inflammation by activating a complex cascade initiated by toxin binding to enterocyte receptors. PMID- 11390764 TI - Visual Pigments and Molecular Genetics of Color Blindness. AB - Red/green color blindness, found in ~1 in 15 men, is caused by the expression of hybrid genes coding for visual pigments. Spectral information from site-directed mutagenesis and recombinant expression has led to the possibility of correlating individual genotypes with psychophysical measurements of the severity of the deficiency. PMID- 11390765 TI - Getting a Grasp on Coordination. AB - Motor coordination is realized by the nervous system at different levels. Concepts about the coordination within a limb define the controlled parameter in cartesian or joint space, whereas concepts about the coordination between limbs stress the temporal control of schemas. Motor deficits of neurological patients point to the role of the different motor areas in motor coordination. PMID- 11390766 TI - Calcium Mobilization Systems During Neurogenesis. AB - Neuroepithelial cells have Ca(2+) mobilization systems that are activated by acetylcholine via muscarinic receptors and by extracellular ATP via P(2U) purinoceptors. The Ca(2+) mobilization occurs during neurogenesis and diminishes in parallel with the declining of mitotic activities of the neuroepithelial cells. Capacitative Ca(2+) influx also occurs with the Ca(2+) mobilization. PMID- 11390767 TI - Transmission at Sympathetic Varicosities. AB - The development of techniques for recording the electrical signs of transmission at single sympathetic varicosities has revealed considerable heterogeneity in the properties of transmission at different varicosities. The origin of these heterogeneities is considered in this short review. PMID- 11390768 TI - Nerves and Hormones Interact to Control Gallbladder Function. AB - Ganglia are the target of several regulatory inputs to the gallbladder. Hormonal cholecystokinin and sympathetic nerves can up- or downregulate neurotransmission in the gallbladder, respectively, by altering the rate of acetylcholine release from vagal preganglionic terminals. Peptides released from sensory axons act directly on gallbladder neurons to increase their excitability. PMID- 11390769 TI - Control of Upper Airway Motoneurons During REM Sleep. AB - The loss of tone in upper airway muscles contributes to disorders of breathing during sleep. In an animal model of rapid eye movement sleep atonia, decrements in the activity of upper airway motoneurons are caused by withdrawal of excitation mediated by serotonin and other transmitters, rather than by state dependent inhibition. PMID- 11390771 TI - Endothelial Gaps: Plasma Leakage During Inflammation. PMID- 11390770 TI - The Lacrimal Gland and Its Veil of Tears. AB - The secretory cells of the lacrimal gland produce a highly complex product of water, ions and proteins. At least five neurotransmitter receptors and three different second message systems are involved in controlling the different secretory processes of this highly sophisticated secretory epithelium. PMID- 11390772 TI - Outer Hair Cells Provide Active Tuning in the Organ of Corti. AB - The detection of sound by the mammalian hearing organ, the organ of Corti, is far from a passive process with the sensory cells acting as mere receptors. The high sensitivity and sharp tuning of the auditory apparatus are very much dependant on the active mechanical behavior of the outer hair cells, acting as effector cells. PMID- 11390773 TI - Maxi-K(Ca), a Unique Member of the Voltage-Gated K Channel Superfamily. AB - Large-conductance, voltage-, and Ca(2+)-sensitive K(+) (maxi-K(Ca)) channels regulate neuronal and smooth muscle excitability. Their pore-forming alpha subunit shows similarities with voltage-gated channels and indeed can open in the practical absence of Ca(2+). The NH(2) terminus is unique, with a seventh transmembrane segment involved in beta-subunit modulation. The long COOH terminus is implied in Ca(2+) modulation. PMID- 11390774 TI - Spatial Memory and Learning in Transgenic Mice: Fact or Artifact? AB - Spatial learning of transgenic mice is often assessed in the Morris watermaze, where mice must use distant cues to locate a submerged platform. Such learning is confounded by species-specific noncognitive swimming strategies. Factor analysis permits cognitive and noncognitive strategies to be disentangled and their association with electrophysiological phenomena to be investigated. PMID- 11390775 TI - The Molecular Physiology of Sodium- and Proton-Coupled Solute Transporters. AB - The expression of cloned Na(+)- and H(+)-coupled solute transporters in Xenopus laevis oocytes has permitted detailed molecular and biophysical analysis and illuminated unique mechanistic features. The identification of missense mutations in inherited diseases and site-directed mutagenesis studies have enhanced our understanding of their roles in physiological and pathological processes. PMID- 11390776 TI - Functional Modulation of Cardiac ATP-Sensitive K(+) Channels. AB - ATP-sensitive K(+) (K(ATP)) channels are inhibited by intracellular ATP, but MgATP is necessary to maintain the channel activity. Numerous cofactors modulate channel function. K(+) channel openers activate and sulfonylureas inhibit K(ATP) channels. The structure of cardiac K(ATP) channel is a complex of mainly K(IR)6.2 and SUR2a. Activation of cardiac K(ATP) channels contributes to action potential shortening during ischemia and plays a role in cardioprotection. PMID- 11390777 TI - The Dichotomy of MIP Family Suggests Two Separate Origins of Water Channels. AB - MIP family proteins can be divided into two groups according to their primary sequences. The CHIP group is predominant in the plant and animal kingdoms and functions primarily as water channels. The GLP group is a minor group with limited prevalence and functions primarily as glycerol transporters. Both prototypes are present in bacteria and may have evolved separately. PMID- 11390778 TI - Reorganization of Somatosensory Cortex After Nerve and Spinal Cord Injury. AB - Somatotopic maps in the mature brain reorganize in response to deafferentation by peripheral nerve cut, amputations, or spinal lesions. Mechanisms underlying these changes may range from altered tonic inhibition and synaptic efficacy to neuronal sprouting. An understanding of these mechanisms could guide interventions that potentiate recovery from such injuries. PMID- 11390780 TI - Adding Endothelium to Artificial Vascular Grafts. PMID- 11390779 TI - Meditation as a Voluntary Hypometabolic State of Biological Estivation. AB - Meditation, a wakeful hypometabolic state of parasympathetic dominance, is compared with other hypometabolic conditions, such as sleep, hypnosis, and the torpor of hibernation. We conclude that there are many analogies between the physiology of long-term meditators and hibernators across the phylogenetic scale. These analogies further reinforce the idea that plasticity of consciousness remains a key factor in successful biological adaptation. PMID- 11390781 TI - In Forthcoming Issue. PMID- 11390782 TI - Capacitative Calcium Entry. AB - Capacitative Ca(2+) entry is a recently discovered Ca(2+) entry pathway that is activated on depletion of intracellular Ca(2+) stores, providing an avenue for store refilling. Despite recent progress in elucidating the capacitative Ca(2+) entry pathway, the mysteries of its molecular identity, its biophysical properties, and the store depletion signal remain. PMID- 11390783 TI - Ethanol: Novel Actions on Nerve Cell Physiology Explain Impaired Functions. AB - Molecular biological tools have revealed receptor proteins for excitatory and inhibitory neurotransmitters on cell membranes as targets of ethanol action. Behavioral and pharmacogenetic assays using rodent lines have supported this neurotransmitter theory of ethanol action and given a firm basis for future identification of the relevant genes and the central physiological processes vulnerable to ethanol. PMID- 11390784 TI - Unraveling the Mystery of Goldblatt Hypertension. AB - In hypertension caused by unilateral renal artery stenosis, the nonstenotic kidney becomes renin depleted but fails to prevent hypertension. The nonstenotic kidney mysteriously develops elevated intrarenal angiotensin II (ANG II) content. Rats chronically infused with ANG II exhibit a similar hypertensive process. The augmentation of intrarenal ANG II is due to receptor-mediated internalization and continued ANG II formation, which provide a hypertensinogenic stimulus. PMID- 11390785 TI - Fast Inactivation of Voltage-Gated K(+) Channels: From Cartoon to Structure. AB - Fast inactivation of voltage-gated potassium (K(v)) channels is the best understood gating transition in ion channels and is brought about by an NH(2) terminal domain (ball domain) of the channel's alpha-subunit, which physically blocks the open pore. Recent analysis by nuclear magnetic resonance spectroscopy showed that ball domains from various K(v) channels exhibit well-defined but distinct structures in aqueous solution. PMID- 11390786 TI - Computer-Aided Design of Thrombin Inhibitors. AB - Computer-aided ligand design is an active, challenging, and multidisciplinary research field that blends knowledge of biochemistry, physics, and computer sciences. Whenever it is possible to experimentally determine or to model the three-dimensional structure of a pharmacologically relevant enzyme or receptor, computational approaches can be used to design specific high-affinity ligands. This article describes methods, applications, and perspectives of computer assisted ligand design. PMID- 11390787 TI - Humoral Regulation of Sleep. AB - Cytokines and hormones, including interleukin-1, tumor necrosis factor, growth hormone-releasing hormone, vasoactive intestinal polypeptide and prolactin, are involved in sleep regulation. These substances enhance sleep, inhibition of them inhibits sleep, and their brain levels vary with sleep. This knowledge helps our understanding of the humoral regulation of sleep. PMID- 11390788 TI - Are There Parallel Channels in the Vestibular Nerve? AB - A popular concept in neurobiology is that sensory information is transmitted to the central nervous system over parallel channels of neurons that play different functional roles. But alternative organizing schemes are possible, and it is useful to ask whether some other framework might better account for the diversity of vestibular primary afferents. PMID- 11390789 TI - In Forthcoming Issue. PMID- 11390790 TI - Renal Water Channels Not Regulated by Vasopressin: How Important Are They to Urine Concentration? PMID- 11390791 TI - The Isolated Perfused Heart and Its Pioneers. AB - In 1866, Carl Ludwig together with Elias Cyon created the first isolated perfused frog heart preparation. Perfusion systems for the isolated mammalian heart were developed by H. Newell Martin in 1883 and by Oscar Langendorff in 1895. In its working mode, the isolated perfused rat heart was established in the 1960s. PMID- 11390792 TI - Calcium Channels Formed by Mammalian Trp Homologues. AB - Homologues of Drosophila trp genes have been isolated from mammalian species in hope that they may constitute the molecular basis of capacitative Ca(2+) entry. Expression of Trps suggests that they form Ca(2+) influx channels regulated by either store depletion or a more upstream event. Store-operated Trp channels can be formed by heteromultimerization. PMID- 11390793 TI - Do Cellular Heat Acclimation Responses Modulate Central Thermoregulatory Activity? AB - The classical concept of heat acclimation is of an autonomically controlled array of integrative physiological processes, improving heat tolerance. New evidence suggests that a temporal interplay between opposing autonomic and peripheral cell originated responses, switched on by heat and autonomic stimulation of the cell membrane, and a marked increase in the stock of the inducible 70-kDa heat shock protein contribute to widening of the thermoregulatory activity span. PMID- 11390794 TI - Two Subgroups of Gonadotropin Releasing Hormone Neurons Control Gonadotropin Secretion in Rats. AB - Two distinct subgroups of gonadotropin-releasing hormone (GnRH) neurons are involved in the control of luteinizing hormone secretion, at least in rats: one subgroup located in the mediobasal hypothalamus constitutes the GnRH pulse generator associated with opioid neurons, and the other located in the preoptic area constitutes the GnRH surge generator associated with gamma-aminobutyric acid neurons. PMID- 11390795 TI - Proteinase-Activated Receptors: New Functions for Old Enzymes. AB - Although proteases are traditionally viewed as degradative enzymes, characterization of a family of G protein-coupled receptors that are activated by proteolysis reveals a new role. Certain proteases function as signaling molecules that specifically regulate cells by cleaving and activating a family of proteinase-activated receptors. PMID- 11390796 TI - Fluid Shear Stress Modulation of Gene Expression in Endothelial Cells. AB - The vascular endothelium, lining the blood vessel wall, is constantly exposed to wall shear stresses generated by flowing blood. Gene regulation, critical for endothelial cell function, depends on complex interactions at the promoter level and utilizes overlapping signal transduction cascades to activate the expression of genes involved in many biological processes. PMID- 11390797 TI - Functional Changes Of Fetal Muscle Acetylcholine Receptor During Mouse Development. AB - In developing muscles in vivo and in vitro, the acetylcholine receptor gamma subunit exists in two splice variants, conferring different single-channel open durations (tau(op)) to reconstituted receptors. In mouse muscles, tau(op) changes around birth, possibly as receptors incorporate either variant of gamma-subunit. This might be relevant to the concomitant maturation of muscle innervation. PMID- 11390799 TI - Ion Channelopathies and Heritable Epilepsy. PMID- 11390798 TI - Flash Photolysis of Caged Compounds: Casting Light on Physiological Processes. AB - Photorelease of bioactive molecules from inactive precursors is a very powerful tool for the study of the molecular mechanisms underlying physiological processes as diverse as ionic channel modulation, exocytosis, phototransduction, ligand receptor interaction, and cross-bridge activity. A brief account of the methodology, available compounds, and fields of application is presented here. PMID- 11390800 TI - In Forthcoming Issue. PMID- 11390802 TI - "Enteric Tears": Chloride Secretion and Its Neural Regulation. AB - Neural reflex circuits within the submucosal plexus and axon reflexes via extrinsic primary afferents control chloride secretion by intestinal epithelial cells. The regulation of chloride secretion is a complex interplay between excitatory and inhibitory influences from neurotransmitters and chemical messengers released from epithelial, endocrine, and immune cells. PMID- 11390801 TI - The Quest for Speed: Muscles Built for High-Frequency Contractions. AB - Vertebrate sound-producing muscles can contract at frequencies greater than 100 Hz, a feat impossible in locomotory muscles. This is not accomplished by unique proteins or structures but by qualitative shifts in isoforms and quantitative reapportionment of structures. Speed comes with costs and trade-offs, however, that restrict how a muscle can be used. PMID- 11390803 TI - Does Your Gut Taste? Sensory Transduction in the Gastrointestinal Tract. AB - The primary sensors in the gut are endocrine cells. They release peptides and amines that stimulate intrinsic and extrinsic neural pathways affecting gastrointestinal motor and secretory function. These regulatory mechanisms alter the digestive and absorptive capacity of the intestine to match the entry of a meal from the stomach. PMID- 11390804 TI - Brain Cooling: An Economy Mode of Temperature Regulation in Artiodactyls. AB - Artiodactyls employ selective brain cooling (SBC) regularly during experimental hyperthermia. In free-ranging antelopes, however, SBC often was present when body temperature was low but absent when brain temperature was near 42 degrees C. The primary effect of SBC is to adjust the activity of the heat loss mechanisms to the magnitude of the heat stress rather than to the protection of the brain from thermal damage. PMID- 11390805 TI - Understanding the Physiology of the Blood-Brain Barrier: In Vitro Models. AB - Endothelial cells exposed to inductive central nervous system factors differentiate into a blood-brain barrier phenotype. The blood-brain barrier frequently obstructs the passage of chemotherapeutics into the brain. Tissue culture systems have been developed to reproduce key properties of the intact blood-brain barrier and to allow for testing of mechanisms of transendothelial drug permeation. PMID- 11390806 TI - In Forthcoming Issue. PMID- 11390807 TI - Regenerative Nerve Fiber Growth in the Adult Central Nervous System. AB - Neurite growth and regeneration in the adult central nervous system (CNS) is extremely limited. An important factor contributing to these restrictions is specific growth inhibitory proteins associated with oligodendrocytes and CNS myelin. A major inhibitory factor is the antigen of a monoclonal antibody; the application of this neutralizing antibody to spinal cord- or brain-lesioned adult rats induces long-distance regeneration of lesioned axons, as well as a specific increase in sprouting and rewiring of the cortical output system to the brain stem and the spinal cord. These anatomic changes are paralleled by important functional recoveries of locomotion and precision movements. PMID- 11390808 TI - Cytokines and Feeding. AB - Cytokines inhibit feeding through peripheral and brain mechanisms. Behavioral, cellular, and molecular studies show that interactions among cytokines, neurotransmitters, and peptides and modulation of hypothalamic neurons are involved in cytokine-induced feeding inhibition. This action of cytokines is relevant to the control of feeding in health and disease. PMID- 11390809 TI - Novel Control of Urokinase Receptor Expression. PMID- 11390810 TI - The Wisdom of the Body Revisited. PMID- 11390811 TI - Healing of Gastrointestinal Mucosa: Involvement of Polyamines. AB - Polyamines are involved in the processes of cell migration and proliferation that result in the repair of mucosal lesions. Depletion of polyamines dramatically alters the arrangement of the cytoskeleton, EGF receptor function, the activities of signal transduction proteins, the levels of several protooncogenes, and the expression and cellular content of at least one growth factor involved in these processes. PMID- 11390812 TI - Chemosensory Perception in the Gut. AB - The ability of the gut mucosa to sense the chemical composition of chyme is important for gastrointestinal functions. The demonstration of gustducin and transducin, two alpha-subunits of GTP-binding proteins involved in gustatory signal transduction, in gastrointestinal epithelial cells provides first clues to the molecular basis of enteric chemosensitivity. Nitric oxide may play a role as a secondary messenger. PMID- 11390813 TI - Bile Acids: The Good, the Bad, and the Ugly. AB - Bile acids, amphipathic end products of cholesterol metabolism, are "good" in the infant because they enhance lipid absorption and thereby promote growth. Bile acids also induce bile flow and biliary lipid secretion. The enterohepatic circulation of bile acids is "bad" in the adult because it downregulates hepatocyte low-density lipoprotein receptor activity and thereby elevates plasma cholesterol levels. Defects in bile acid metabolism such as impaired biosynthesis or transport are "ugly" because they cause morbidity and death. New approaches for treating these defects are being developed. PMID- 11390814 TI - Nitric Oxide and Body Temperature Control. AB - Pharmacological studies of thermoregulatory effector and neuronal responses indicate that nitric oxide (NO) may have differential roles in the control of body temperature and during fever. Histochemical analysis of site-specific changes in NO synthase activity in defined states of thermal stimulation appears a promising approach to unravel the underlying hypothalamic neuronal cytoarchitecture. PMID- 11390816 TI - Regulation of Cellular Growth by 1,25-Dihydroxyvitamin D(3)-Mediated Growth Factor Expression. AB - Although the primary function of 1,25-dihydroxyvitamin D(3) [1,25(OH)(2)D(3)] is the regulation of calcium and phosphorus metabolism, it also regulates the growth and differentiation of several different cell types. Recent work suggests that 1,25(OH)(2)D(3) regulates cellular growth by altering the synthesis of growth factors and growth factor responses. PMID- 11390817 TI - Molecular Features of Energy Coupling in the F(0)F(1) ATP Synthase. AB - H(+) translocation is coupled to ATP synthesis in the F(0)F(1) ATP synthase via a rotary mechanism. Catalytic turnover, site-site cooperativity, and H(+) transport obligatorily involve rotation of a set of subunits. The transport domain in the membranous F(0) and the catalytic domain in the F(1) are mechanisms designed for generating torque. PMID- 11390818 TI - Urea Transport: It's Not Just "Freely Diffusible" Anymore. PMID- 11390819 TI - The Dual Role of Nitric Oxide in Islet beta-Cells. AB - In pancreatic islets, nitric oxide (NO) produced on exposure to cytokines mediates beta-cell injury leading to diabetes mellitus. On the other hand, L arginine-derived NO may participate in the signal transduction pathway of physiological insulin secretion. This review focuses on the dual role of NO as a mediator of physiological and pathophysiological processes in pancreatic islets. PMID- 11390820 TI - Toward Multifactorial Hypothalamic Regulation of Anterior Pituitary Hormone Secretion. AB - The hypothalamus regulates the secretion of anterior pituitary hormones via release of releasing hormones into the hypophysial portal vasculature. Additional neuromessengers act at the pituitary to modulate responses to the hypothalamic hormones. For example, neuropeptide Y enhances the effect of gonadotropin releasing hormone and the response to the prolactin-inhibiting hormone dopamine. PMID- 11390821 TI - SCFAs: The Enigma of Weak Electrolyte Transport in the Colon. AB - Short-chain fatty acids are the predominant luminal anions in the colon (>75 mM) and thus create a rather unique environment for transporting epithelium. The colon absorbs short-chain fatty acids, either by diffusion of the protonated species across the apical membrane or by an anion exchange process with bicarbonate. Additionally, short-chain fatty acids modulate Na absorption, Cl secretion, intracellular pH, and cell volume. PMID- 11390822 TI - News in Motor Cortical Physiology. AB - Motor cortical activity relates to static motor parameters (isometric force, limb position) under static conditions but predominantly to dynamic parameters (change of force, limb velocity) under dynamic conditions. This dual relation conceptually unifies the role of motor cortex in the control of isometric force and movement. PMID- 11390824 TI - A Unique Role of NO in the Control of Blood Flow. AB - Nitric oxide synthase (NOS) inhibitors induce significant vasoconstriction, suggesting an indispensable role of NO as a local vasodilator. This is due mainly to its effects on large arterioles that significantly control arterial conductance while scarcely being regulated by metabolites. NO's role in adapting vascular conductance to flow is pronounced during (re)active hyperemia and autoregulation. PMID- 11390823 TI - Information Networks in the Arterial Wall. AB - The main task of the arterial system is to secure an adequate supply of oxygen to organs. This fact implies the integration of multiple signals in the vascular wall. This review deals with the exchange of information between and among smooth muscle and endothelial cells through gap junctions in the vessel walls of arteries and arterioles. PMID- 11390825 TI - Renal Transport of Urate in Humans. AB - The theory of the "four-component model" of urate excretion in humans is reevaluated, considering that a decrease in urate excretion induced by drugs like pyrazinamide or by endogenous compounds like lactate and ketone bodies might be a result of stimulation of urate reabsorption and not, as previously considered, of inhibition of urate secretion. PMID- 11390826 TI - Metaplasticity: Key Element in Memory and Learning? PMID- 11390828 TI - An Integrated View of Insect Flight Muscle: Genes, Motor Molecules, and Motion. AB - Substituting an alanine for serine in the regulatory subunit of the motor protein myosin dramatically alters Drosophila's flight ability. Power output, at all levels of the flight system, is reduced. This example of deciphering a protein's function by producing malfunctions illustrates the broadening use of molecular genetics in integrative biology. PMID- 11390829 TI - Epithelial Fluid Transport--A Century of Investigation. AB - Our current understanding of the mechanism of fluid transport by epithelia relies upon mathematical models developed 30 years ago to explain the mechanism of solute-solvent coupling and the pathways taken by water across an epithelium. The validity of these models is reconsidered in light of recent findings, and it is concluded that a simple three-compartment model system is adequate to explain fluid absorption and secretion by epithelia. PMID- 11390830 TI - Commentary on "Epithelial Fluid Transport--A Century of Investigation" PMID- 11390831 TI - Central Neuronal Histamine Contributes to Cardiovascular Regulation. AB - Evidence suggests that central histaminergic neurons make important contributions to cardiovascular regulation. For example, histamine-containing neurons project to brain regions important for cardiovascular regulation. Furthermore, stimulation of central histamine receptors changes blood pressure and heart rate and alters activity of major vasoconstrictor systems. Finally, histamine metabolism is changed in hypertension. PMID- 11390832 TI - Stimulation of Glucose Transport by Hypoxia: Signals and Mechanisms. AB - Glucose transport is acutely stimulated by hypoxia through enhanced GLUT-1 and GLUT-4 glucose transporter function. GLUT-1 expression is also stimulated by hypoxia or azide. Moreover, hypoxia per se, acting through hypoxia-inducible factor 1, enhances GLUT-1 transcription. GLUT-1 is the first gene whose transcription is dually stimulated in response to hypoxia and inhibition of oxidative phosphorylation. PMID- 11390833 TI - The Pattern of Sympathovagal Balance Explored in the Frequency Domain. AB - In most physiological conditions, sympathetic and vagal activities modulating heart period undergo a reciprocal regulation, leading to the concept of sympathovagal balance. This pattern can be indirectly quantified by computing the spectral powers of the oscillatory components corresponding to respiratory acts (high frequency) and to vasomotor waves (low frequency) present in heart rate variability. PMID- 11390834 TI - The Role of the Sinusoidal Endothelium in Liver Function. AB - Microvascular exchange in the liver is governed by fenestrations in sinusoidal endothelial cells and can be manipulated pharmacologically. Microvascular exchange is affected in alcoholic liver disease and cirrhosis, the former leading to a loss of fenestrae, the latter to sinusoidal capillarization and thereby to loss of liver function in disease. PMID- 11390835 TI - Arteriogenesis Versus Angiogenesis: Two Mechanisms of Vessel Growth. AB - After birth, new blood vessel formation proceeds via angiogenesis or arteriogenesis. Angiogenesis (capillary sprouting) results in higher capillary density. Arteriogenesis (rapid proliferation of collateral arteries) is potentially able to significantly alter the outcome of coronary and peripheral artery disease. The processes share some growth features but differ in many aspects. PMID- 11390837 TI - Altered NaCl Concentration of Airway Surface Liquid in Cystic Fibrosis. PMID- 11390838 TI - Ca(2+) Sparks in Cardiac Muscle: Is There Life Without Them? AB - The discovery of elementary Ca(2+) signaling events, the Ca(2+) sparks, has profoundly changed our view of cardiac Ca(2+) signaling and, in particular, excitationcontraction coupling. In addition, a partial disruption of cardiac Ca(2+) signaling may be important in clinical cardiac conditions such as congestive heart failure. Understanding cardiac diseases on the cellular and molecular levels may provide a key to new pharmacological strategies in the near future. PMID- 11390839 TI - The ether-a-go-go-Related Gene K(+) Current: Functions of a Strange Inward Rectifier. AB - The erg channels mediate an inward-rectifying K(+) current because of their peculiar gating kinetics. They are involved in repolarization of the cardiac action potential, frequency adaptation, and maintenance of the resting potential. Reduction of erg currents via an intracellular signal cascade underlies the thyrotropin-releasing hormone-induced increase in prolactin secretion. PMID- 11390840 TI - Probing Nanometer Structures with Atomic Force Microscopy. AB - Atomic force microscopy (AFM) can generate high-resolution images of the surface of biological specimens and can also probe the interactions between and within single macromolecules. Thus isolated heterogeneous biological structures can be studied in submolecular detail with AFM. PMID- 11390841 TI - Hepatic Regeneration-Revisiting the Myth of Prometheus. AB - Myriad signals such as growth factors, cytokines, growth inhibitors, hormones, ions, extracellular matrix, and the resident hepatic cells are involved in the regulation of hepatic regeneration. These regulatory factors ultimately mediate changes in gene expression, a critical step in this well-orchestrated restorative process. PMID- 11390842 TI - Advances in Parasympathetic Control of Heart Rate and Cardiac Function. AB - It is well known that the neuronal projections from the brain to the heart strongly influence cardiac function, and an abnormal activity has been implicated in diseases such as cardiac arrhythmia and sudden infant death syndrome. This short review describes recent advances focused on the neurobiology of cardiac vagal neurons, utilizing cellular techniques. PMID- 11390843 TI - Subcellular Ca(2+) Dynamics. AB - The field of subcellular Ca(2+) homeostasis is evolving rapidly. In parallel with improvements in spatial and temporal resolution of Ca(2+) imaging techniques, new methods using the natural cell machinery to target Ca(2+)-sensitive proteins such as aequorin to precise intracellular locations promise superb specificity to measure [Ca(2+)] in defined subcellular environments. PMID- 11390844 TI - Motor Patterns in Walking. AB - Despite the fact that locomotion may differ widely in mammals, common principles of kinematic control are at work. These reflect common mechanical and neural constraints. The former are related to the need to maintain balance and to limit energy expenditure. The latter are related to the organization of the central pattern-generating networks. PMID- 11390846 TI - ER Muscles Its Way Around Neurons. PMID- 11390847 TI - Astrocytes Couple Synaptic Activity to Glucose Utilization in the Brain. AB - Astrocytes have functional characteristics that make them particularly well suited to couple glutamate uptake from the synaptic cleft to Na(+)-K(+)-ATPase activation and glucose utilization. The changes in glucose metabolism associated with these processes may provide signals detected by positron emission tomography. PMID- 11390849 TI - Aquaporins: Phylogeny, Structure, and Physiology of Water Channels. AB - How water permeates cellular membranes and what this means for cell functioning and several diseases are now emerging from the study of the aquaporins (AQPs), the water channel family. A combination of sequence analysis, three-dimensional structure determination, and physiology of the AQP family proteins provides a glimpse into the workings of water channels. PMID- 11390848 TI - Vascular structure determines pulmonary blood flow distribution. AB - Scientific knowledge develops through the evolution of new concepts. This process is usually driven by new methodologies that provide observations not previously available. Understanding of pulmonary blood flow determinants advanced significantly in the 1960s and is now changing rapidly again, because of increased spatial resolution of regional pulmonary blood flow measurements. PMID- 11390851 TI - Visceral Afferent Neurons: Role in Gastric Mucosal Protection. AB - Gastric mucosal homeostasis requires rapid alarm of protective mechanisms in the face of pending injury. This article summarizes the evidence that spinal afferent neurons monitor insults to the gastric mucosa and activate local mechanisms of defense and repair through release of transmitter peptides from their endings in the stomach. PMID- 11390850 TI - Physiology of Receptor-Mediated Lymphocyte Apoptosis. AB - Apoptosis (programmed cell death), a physiological mechanism eliminating abundant and potentially harmful cells, is triggered by a variety of stimuli including activation of distinct receptors. The machinery mediating CD95 receptor-induced apoptosis includes caspases, ceramide, kinases, Ras and Rac, formation of O(2)( ), mitochondrial proteins, inhibition of K(+) channels, activation of Cl(-) channels, and osmolyte release. PMID- 11390852 TI - Spatiotemporal Dynamics of Human Cognition. AB - It is still largely unknown how the complex cortical neural network of the human brain can process information so rapidly. Multichannel evoked potential recordings with millisecond time resolution and spatiotemporal analysis methods now allow us to address this question and to unravel the temporal dynamics of the large-scale neurocognitive networks. PMID- 11390854 TI - G Protein Regulation of Inwardly Rectifying K(+) Channels. AB - Inwardly rectifying K(+) (Kir) channels respond to receptor-stimulated signaling cascades that involve G proteins and other cytosolic messengers. Channel activity is controlled both by direct coupling of G protein subunits and by phosphorylation via protein serine/threonine and tyrosine kinases. The coincidence of both forms of Kir channel signaling may give rise to complex cellular responses. PMID- 11390855 TI - Escape from Vasopressin-Induced Antidiuresis: Insights at the Molecular Level. PMID- 11390856 TI - Prolactin, the Hormone of Paternity. AB - Prolactin has long been known to play a significant role in maternal care. When behavioral endocrinologists began to examine the endocrinology of fatherhood, prolactin was also found to be connected with paternal care in fish, birds, and mammals including primates. PMID- 11390857 TI - Sensorimotor Integration at the Dorsal Column Nuclei. AB - Interaction among primary afferents, corticofugal fibers, and intrinsic elements allows for sensorimotor integration at the dorsal column nuclei. The interneurons permit the spatial localization, the recurrent collaterals synchronize the activity of projecting cells with overlapping receptive fields, and the corticofugal fibers induce a central zone of activity surrounded by a peripheral zone of inhibition. PMID- 11390858 TI - P-450 Eicosanoids: A Novel Signaling Pathway Regulating Renal Function. AB - Cytochrome P-450 enzymes primarily metabolize arachidonic acid to epoxyeicosatrienoic acids (EETs) and 20-hydroxyeicosatetraenoic acid (20-HETE) in the kidney. These compounds serve as second messengers that play a central role in the regulation of renal vascular tone and sodium reabsorption in the proximal tubule and thick ascending loop of Henle. PMID- 11390859 TI - PTH and PTHrP: Similar Structures but Different Functions. AB - Parathyroid hormone and parathyroid hormone-related peptide are two structurally similar peptide hormones that exert mainly identical effects on classical target cells of parathyroid hormone but remarkable differences in their effects on other target cells. This review focuses on their functional differences and the mechanisms underlying these effects. PMID- 11390860 TI - The Reward Signal of Midbrain Dopamine Neurons. AB - Dopamine projections from the midbrain to striatum and frontal cortex play a major role in behavioral reactions controlled by rewards. Recent experiments have shown that dopamine neurons code the discrepancy between the prediction and occurrence of rewards and in this way signal a crucial learning term for approach behavior. PMID- 11390861 TI - Adrenomedullin: Is There Physiological Relevance in the Pathology and Pharmacology? AB - The adrenomedullin gene encodes two potent hypotensive peptides, adrenomedullin and proadrenomedullin NH(2)-terminal 20 peptide. As with other vasoactive peptides, the most difficult challenge is to prove the physiological relevance of their recognized pathological and pharmacological actions and to establish, against that physiological background, their therapeutic potential. PMID- 11390862 TI - From Limbs to Lungs: A Newt Perspective on Compensatory Lung Growth. AB - Partial lung resection initiates compensatory growth of remaining lobes to restore pulmonary structure and function. Mechanisms underlying this response are not well defined. This article considers molecular pathways involved in control of amphibian limb regeneration and tissue pattern formation for novel insight into the understanding of compensatory lung growth. PMID- 11390864 TI - The Discovery of Renin 100 Years Ago. AB - In 1898, Tigerstedt and Bergman published their observation that kidney extracts produce pressor effects. They characterized the substance and named it "renin." Although this was the beginning of understanding the role of the kidney in hypertension and the renin-angiotensin system, their discovery lay dormant for nearly 40 years. PMID- 11390865 TI - Sgk: A New Player (Star?) in the Early Action of Aldosterone. PMID- 11390867 TI - Endothelium-Derived Hyperpolarizing Factor-Fact or Fiction? AB - The vascular endothelium releases a diffusible factor that hyperpolarizes and hence relaxes vascular smooth muscle cells predominantly through activation of Ca(2+)-dependent K(+) channels. In the coronary circulation, this endothelium derived hyperpolarizing factor appears to be a cytochrome P(450)-derived arachidonic acid epoxide, the release of which may play a crucial role in the maintenance of coronary blood flow in arteriosclerosis. PMID- 11390868 TI - Bile Formation: a Concerted Action of Membrane Transporters in Hepatocytes and Cholangiocytes. AB - A large number of membrane transport mechanisms in hepatocytes and cholangiocytes serves for the secretion of bile acids, various other organic anions, organic cations, lipids, and electrolytes. After their functional characterization, some of these mechanisms' individual transport molecules are now identified, allowing better understanding of inherited and acquired disorders of bile formation. PMID- 11390869 TI - Ceramides as Key Players in Cellular Stress Response. AB - The recent discovery of sphingolipid-derived second messengers that regulate fundamental cell responses such as cell growth and apoptosis has provided insight into the way cells sense and respond to stressful stimuli. This will help the understanding of the pathogenesis of stress-related diseases and eventually offer novel therapeutic approaches. PMID- 11390870 TI - Extracellular Surface Charges in Voltage-Gated Ion Channels. AB - A large number of charged amino acids are in the extracellularly located parts of the voltage-gated ion channels. Recent findings suggest that these surface charges contribute to the channel functions in the sensing of voltage, the binding of substances, and the sensing of H(+) concentration. PMID- 11390871 TI - Measurement and Interpretation of Cytoplasmic AB - Ca(2+)-indicator dyes are widely used in biology yet difficult to characterize inside cells. Studies in skeletal muscle fibers provide important information about indicator behavior and about Ca(2+) signaling within the cytoplasm. PMID- 11390872 TI - Sarcomeric Myosin Isoforms: Fine Tuning of a Molecular Motor. AB - Sarcomeric or striated muscle myosins are the molecular motors whose fine tuning is best known. Sarcomeric myosin isoforms convert chemical into mechanical energy at very different rates without losing efficiency. Availability of the amino acid sequences offers for the first time the chance to understand the molecular basis of the versatility of these molecular motors. PMID- 11390873 TI - Images of Action Potential Propagation in Heart. AB - Activation and repolarization across mammalian hearts follow complex three dimensional pathways that are governed by fiber structure, intercellular coupling, and action potentials (APs) with spatially heterogeneous properties. Voltage-sensitive dyes and imaging techniques offer new insights on how spatiotemporal heterogeneities of APs govern propagation, repolarization, and AV node conduction and help us visualize arrhythmias with previously unattainable details. PMID- 11390874 TI - Phospholipase C Isoforms, Cytoskeletal Organization, and Vascular Smooth Muscle Differentiation. AB - The function of differentiated vascular smooth muscle cells (VSMC) in vivo is the regulation of contractility. Following injury or disease, however, VSMC lose their contractile function and take on a synthetic, proliferative phenotype. This dedifferentiation is generally accompanied by a change in the expression profile of phospholipase C isoforms. PMID- 11390875 TI - Presynaptic Heterogeneity: Vive la difference. AB - Since individual synapses of the same neuron may have different molecular composition, an important question in neurobiology is how the properties of individual synapses are established and maintained. Recent technical advances allow assay of activity at individual synapses and investigation of the relationship between function and molecular composition at the synapse. PMID- 11390877 TI - Cytosolic Calcium Oscillations in Smooth Muscle Cells. AB - In a variety of smooth muscle cells, agonists activating membrane receptors induce oscillations in the cytoplasmic Ca(2+) concentration via an inositol trisphosphate-activated mechanism. Ca(2+) oscillations participate in the control of cell membrane potential and the tone of smooth muscle. There is evidence that alterations in Ca(2+) oscillations modulate smooth muscle responsiveness. PMID- 11390878 TI - Determination of Left-Right Asymmetry: Role of Cilia and KIF3 Motor Proteins. PMID- 11390879 TI - CFTR Chloride Channels: Binding Partners and Regulatory Networks. AB - The cystic fibrosis gene encodes a chloride channel (CFTR) that regulates transepithelial salt and water transport. Two classes of CFTR-binding proteins appear to link the opposing cytoplasmic tails of this channel to distinct regulatory networks. Such interactions may constitute new paradigms for modulating CFTR activity in health and disease. PMID- 11390880 TI - Interface Properties of Circumventricular Organs in Salt and Fluid Balance. AB - The "sensory" circumventricular organs, with their leaky blood-brain barriers, permit contact between brain neurons and blood-borne molecules. Body fluid balance and cardiovascular control involve established interface functions of subfornical organs. Their recently identified target functions for hormones released during digestion suggest that they may coordinate fluid and food intake. PMID- 11390881 TI - Intracellular Virus Trafficking Reveals Physiological Characteristics of the Cytoskeleton. AB - Virus particles that infect eukaryotic cells can take advantage of the cytoskeleton and associated motors to translocate through the cytoplasm. Depending on the virus, motor proteins are recruited or, alternatively, cytoskeletal elements are induced to polymerize onto viral structures. Here we review recent advances toward understanding the roles of the cytoskeleton in virus trafficking. PMID- 11390882 TI - The Cell Physiology of Biphasic Insulin Secretion. AB - Glucose-stimulated insulin secretion consists of a transient first phase followed by a sustained second phase. Diabetes (type II) is associated with abnormalities in this release pattern. Here we review the evidence that biphasic insulin secretion reflects exocytosis of two functional subsets of secretory granules and the implications for diabetes. PMID- 11390883 TI - Mechanics of Relaxation of the Human Heart. AB - Rapid and complete relaxation is a prerequisite for cardiac output adaptation to changes in loading conditions, inotropic stimulation, and heart rate. In the healthy human heart, the rate and extent of relaxation depend mainly on actomyosin cross bridge dissociation and on left ventricular end-systolic volume, rather than on the afterload level. PMID- 11390884 TI - The Time Course of Signaling at Central Glutamatergic Synapses. AB - Glutamate is the main excitatory transmitter in the mammalian CNS, mediating fast synaptic transmission primarily by activation of AMPA-type glutamate receptor channels. Both synaptic structure and a cell-specific molecular switch in the AMPA receptor subunit expression are involved in the regulation of the synaptic signaling time course. PMID- 11390885 TI - Molecular Mechanisms in Bile Formation. AB - Active canalicular secretion of bile salts and non-bile salt organic anions represents the major driving force of hepatic bile formation. The most important carriers involved have now been cloned on both the basolateral and canalicular sides of hepatocytes. Elucidation of their structure, transport properties, and regulation is an important step forward in the ultimate understanding of the molecular physiology of bile formation. PMID- 11390886 TI - Brain Stem Reflexes: Probing Human Trigeminal Nociception. AB - Although many people suffer from orofacial pain and headache, objective methods for investigation of trigeminal nociception in humans have been lacking. Trigeminal brainstem reflexes such as the masseter inhibitory reflex and the blink reflex are mediated by central multireceptive neurons that are also involved in trigeminal nociception. Therefore, these trigeminal reflexes are suitable models for probing pontine and medullary pain processing. PMID- 11390887 TI - Synaptic Transmission at Single Boutons in Sympathetic Ganglia. AB - Synaptic transmission has traditionally been studied at the level of the entire nerve terminal rather than at one of its constituent boutons. Autonomic ganglia provide preparations for recording from individual boutons, as well as for determining the calcium transients necessary for transmitter release at these boutons. The results suggest a new paradigm for synaptic transmission. PMID- 11390888 TI - Breathless Legs? Consider Training Your Respiration. AB - The condition of the respiratory system is more important for endurance exercise performance of healthy subjects than hitherto assumed. Not only do respiratory muscles fatigue during intensive endurance exercise, but prefatigued respiratory muscles can also impair performance. In turn, respiratory endurance training can improve endurance exercise performance. PMID- 11390890 TI - Possible Role of Ion Channels in the Action of Nonsteroidal Anti-Inflammatory Drugs. PMID- 11390891 TI - Adhesion Molecules: The Path to a New Understanding of Acute Inflammation. AB - About 100 years after the definition of the basic principles of inflammation, the identification of the underlying molecular mechanisms provides a new understanding of the inflammatory response. The specificity and diversity of the adhesion molecules involved in leukocyte extravasation account for the ordered leukocyte recruitment and activation in inflammation. PMID- 11390892 TI - Is the GI System Built For Exercise? AB - The gut usually meets the fluid, electrolyte, and nutrient requirements of mild to heavy exercise. Gastric emptying and intestinal absorption rates of beverages ingested during exercise equal sweat rates. However, strenuous or prolonged exercise under dehydrated conditions can produce gastrointestinal distress and tissue damage. PMID- 11390893 TI - Apoptosis in the Retina: The Silent Death of Vision. AB - Pathogenetic mechanisms of retinal degeneration include cell loss by apoptosis. This gene-regulated mode of single-cell death occurs in a number of widespread human diseases such as neurodegeneration. The knowledge of genes and signaling in retinal apoptosis is expanding and opens up therapeutic strategies to ameliorate blinding retinal diseases. PMID- 11390894 TI - The Beauty and the Beast: Aspects of the Autonomic Nervous System. AB - Sympathetic nerve activity is altered and is a prognostic factor for many cardiovascular diseases such as hypertension, coronary syndromes, and congestive heart failure. Therefore, the selection of vasoactive drugs for the treatment of these diseases should also take into consideration their effects on the sympathetic nervous system. PMID- 11390895 TI - Exuberant Ca(2+) Signaling in Neutrophils: A Cause for Concern. AB - Under inflammatory conditions after burn/trauma injuries, circulating neutrophils are frequently hyperactive, contributing to excessive superoxide production and related tissue damage. Although normal neutrophil activation is cooperatively controlled by Ca(2+)-independent and Ca(2+)-linked signaling pathways, exuberant Ca(2+)-linked signaling appears to cause neutrophil hyperactivation in the injury conditions. PMID- 11390896 TI - Key Roles of Renal Aquaporins in Water Balance and Water-Balance Disorders. AB - The discovery of aquaporins by Agre and co-workers provided an answer to the long standing biophysical question of how water can pass cell membranes. The identification and characterization of several aquaporins expressed in the kidney has allowed detailed insight, at the molecular level, into the fundamental physiology and pathophysiology of renal water metabolism. PMID- 11390897 TI - Atrial Natriuretic Peptide: Regulator of Chronic Arterial Blood Pressure. AB - Recent findings in atrial natriuretic peptide (ANP) transgenic and gene knockout mouse models uncovered a tonic vasodilatory effect of this hormone that contributes to chronic blood pressure homeostasis. With elevated salt intake, ANP mediated antagonism of the renin-angiotensin system is essential for blood pressure constancy, suggesting that a deficiency in ANP activity may underlie the etiology of sodium-retaining disorders. PMID- 11390898 TI - Regulation of Mammalian Gene Expression by Glucose. AB - Recent data suggest that cells from species as diverse as yeast and mammals may use similar mechanisms to detect changes in nutrient concentration. Here we review recent advances in understanding how glucose regulates gene transcription in mammals. PMID- 11390900 TI - SOC and unSOC. PMID- 11390899 TI - Stimulatory and Inhibitory Functions of the R Domain on CFTR Chloride Channel. AB - CFTR is a chloride channel whose gating process involves coordinated interactions among the regulatory (R) domain and the nucleotide-binding folds (NBFs). Protein kinase A phosphorylation of serine residues renders the R domain from inhibitory to stimulatory and enables ATP binding and hydrolysis at the NBFs, which in turn control opening and closing of the chloride channel. PMID- 11390901 TI - Toward Understanding the Role of Methylation in Aldosterone-Sensitive Na(+) Transport. AB - Proper endocrine regulation of Na(+) reabsorption by renal principal cells is the primary means in mammals for maintaining blood pressure. Aldosterone increases Na(+) reabsorption by activating luminal Na(+) channels; however, the signal transduction pathway of aldosterone is not fully understood. Cellular methylation is necessary for aldosterone signaling to the luminal Na(+) channel. We describe the enzymes, regulators, and effectors of aldosterone-mediated methylation relevant to Na(+) reabsorption. PMID- 11390902 TI - Ca(2+)-Activated Cl(-) Channels: A Newly Emerging Anion Transport Family. AB - A new family of Cl(-) channels widely expressed in epithelia has been identified. These proteins are associated with Ca(2+)-sensitive conductive Cl(-) transport when heterologously expressed. This family may underlie the Ca(2+)-mediated Cl(-) conductance responsible for rescue of the cystic fibrosis knockout mouse from significant airway disease. PMID- 11390903 TI - Angiotensin II Receptor Physiology Using Gene Targeting. AB - The study of mice with targeted disruptions of angiotensin receptor genes has provided new insights into the roles of the individual receptor subtypes, i.e., AT(1A), AT(1B), and AT(2), in growth, development, and the regulation of blood pressure. PMID- 11390904 TI - Isoform-Specific Regulation of the Na(+)-K(+) Pump in Heart. AB - Guinea pig ventricular myocytes coexpress two isoforms of the Na(+)-K(+) pump. These two isoforms respond differently to the physical environment and are coupled to autonomic input through different signal transduction cascades. The expression of different isoforms provides each cell type with a mechanism of programming specific responses to environmental changes. PMID- 11390905 TI - How a Turtle's Shell Helps It Survive Prolonged Anoxic Acidosis. AB - Anoxic turtles accumulate high levels of lactate in blood. To avoid fatal acidosis, turtles exploit buffer reserves in their large mineralized shell. The shell acts by releasing calcium and magnesium carbonates and by storing and buffering lactic acid. Together with profound metabolic depression, shell buffering permits survival without oxygen for several months at 3 degrees C. PMID- 11390906 TI - Mechanisms of Modulation of a Neural Network. AB - Neural networks form the basis for the generation and control of various patterns of behavior. Such networks are subjected to modulatory systems that influence their operation and, thereby, the behavior. In the lamprey locomotor network, analysis on the ion channel, synaptic, and cellular levels has given new insights into the organization of such modulatory systems. PMID- 11390908 TI - Marshaling the Anti-Inflammatory Influence of the Neuroimmunomodulator alpha-MSH. AB - Inflammation in peripheral tissues should benefit from a novel focus on descending neuroimmunomodulatory anti-inflammatory influences stemming from CNS receptors for the peptide alpha-MSH. Because activation of glial and peripheral cell receptors modulates inflammatory activity, alpha-MSH peptides may aid treatment of a variety of CNS and peripheral inflammatory disorders. Activation of melanocortin receptors should thus promote control of inflammatory diseases. PMID- 11390909 TI - Testicular Estrogens and Male Reproduction. AB - Besides somatic cells, aromatase gene expression and its transduction in an active protein in germ cells provides evidence of an additional site for estrogen production within testes of some mammals. Together with the widespread distribution of estrogen receptors in testicular cells, these data illuminate the hormonal regulation of male reproductive function PMID- 11390910 TI - Autoregulation of Glucose Production. AB - Glucose itself regulates endogenous glucose production independently of changes in glucoregulatory hormones. In addition, acute stimulation of gluconeogenesis does not increase net glucose production. This indicates autoregulation of glucose production. Glucokinase plays a role in this process by allowing hepatic glucose sensing. PMID- 11390911 TI - Oxygen Radicals as Messengers in Oxygen-Dependent Gene Expression. AB - Changes in ambient PO(2) need to be sensed to allow long-term adaptation of cellular functions via the regulation of gene activity. The generation of OH* from H(2)O(2) in an iron-dependent perinuclear Fenton reaction for triggering gene expression could be the key event in the O(2) signaling pathway. PMID- 11390912 TI - The 34th Congress of the International Union of Physiological Sciences. PMID- 11390913 TI - Mast Cell-Nerve Interactions. AB - Mutual associations between nerves and mast cells have been observed in normal conditions and in pathological ones such as human irritable bowel syndrome, atopic dermatitis, interstitial cystitis, and more. Here we review the recent literature in this field, putting emphasis on the enteric, skin, and urinary systems, and the pathophysiological implications of this interaction in them. PMID- 11390914 TI - The Organization of the Frontal Motor Cortex. AB - Recent anatomic and functional data radically changed our ideas about the organization of the motor cortex in primates. Contrary to the classic view, the motor cortex does not consist of two main areas, primary and supplementary motor areas, but of a mosaic of cortical areas with specific connections and functional properties. PMID- 11390915 TI - Neuroprotection and Angiogenesis: Dual Role of Erythropoietin in Brain Ischemia. AB - Erythropoietin, originally defined as an erythroid growth factor, is upregulated in the brain under conditions of hypoxia. So far, two functions have been identified for this locally produced cytokine: a direct protective effect on neuronal cells during cerebral ischemia and an indirect protection of brain tissue that could be provided by promoting brain vessel growth. PMID- 11390916 TI - Role of Nitric Oxide in Buffering Short-Term Blood Pressure Fluctuations. AB - Blood pressure instability may promote cardiovascular morbidity. Recent data suggest a role of nitric oxide in stabilizing arterial blood pressure. A rise in blood pressure enhances endothelial shear stress and nitric oxide release. The resulting vasodilation antagonizes the initial increase in blood pressure. This system can respond within 2-10 seconds. PMID- 11390917 TI - Neurophysiology of Spatial Cognition. AB - Understanding of the neurophysiology of cognition is advancing through the study of how animals navigate and understand space. Manipulating various classes of spatial information and recording from hippocampal neurons provides a robust model for understanding how the brain stores and constructs the spatial memories that are critical for organizing daily experience. PMID- 11390918 TI - Stopping Exercise: Role of Pulmonary C Fibers and Inhibition Of Motoneurons. AB - In animals, the J reflex evoked by pulmonary C fibers provides potent inhibition of limb muscles and would act to limit exercise. However, recent work shows that although activation of these fibers causes severe respiratory discomfort, it does not impair the output of limb motoneurons to voluntary, reflex, or locomotor drives in awake humans. PMID- 11390919 TI - Melatonin: Lowering the High Price of Free Radicals. AB - The endogenous antioxidative defense system reduces molecular toxicity of oxygen and nitrogen-based reactive species. Melatonin is an efficient direct and indirect antioxidant. It detoxifies the highly reactive hydroxyl radical and neutralizes other toxic species, including singlet oxygen, hydrogen peroxide, nitric oxide, and peroxynitrite anion, and stimulates several antioxidative enzymes. PMID- 11390920 TI - The Neuroimmune Interface in Prion Diseases. AB - Prion diseases are fatal neurodegenerative disorders of animals and humans. Here we address the role of the immune system in the spread of prions from peripheral sites to the central nervous system and its potential relevance to iatrogenic prion disease. PMID- 11390921 TI - The Role of Dystroglycan and Its Ligands in Physiology and Disease. AB - Dystroglycan contributes to the formation of basement membrane during embryonic development and enforces cell membrane integrity by bridging cytoskeleton and components of the extracellular matrix. In several forms of muscle disease, dystroglycan is reduced in abundance. Moreover, human viral and bacterial pathogens use dystroglycan as their cellular entry point. PMID- 11390922 TI - Tryptophan-Free Diets: A Physiological Tool to Study Brain Serotonin Function. AB - Tryptophan-free diets produce a specific reduction of brain serotonin synthesis and release. This method for lowering neural serotonin function has been extensively used in both laboratory animals and humans to study the role of serotonin in a variety of behaviors, such as aggressiveness, sleep, sexual behavior, anxiety, mood, memory, and so forth. PMID- 11390923 TI - Urocortin: A Novel Player in Cardiac Control. AB - Urocortin is a potent regulator of cardiac function, with actions that are prolonged in experimental animals. These changes are mediated via binding to corticotropin-releasing factor receptors found in peripheral tissues. The effects of urocortin on behavior, appetite, inflammation, and the cardiovascular system suggest that this peptide may be an endogenous factor mediating actions previously attributed to corticotropin-releasing factor. PMID- 11390925 TI - Myoglobin: Just an Oxygen Store or Also an Oxygen Transporter? AB - Besides acting as an oxygen store during times of reduced blood oxygen supply, myoglobin can also facilitate intracellular oxygen transport by diffusion of oxymyoglobin along a PO(2) gradient. We reassess the importance of myoglobin facilitated oxygen diffusion by applying new findings on the intracellular diffusivity of myoglobin in a model calculation. PMID- 11390926 TI - No Role for the Ryanodine Receptor in Regulating Cardiac Contraction? AB - Cardiac contraction is initiated by Ca(2+) leaving the sarcoplasmic reticulum through the ryanodine receptor (RyR). Although opening of the RyR can be modified by various ligands, these have no maintained effect on contraction. We conclude that modulation of the RyR controls sarcoplasmic reticulum Ca(2+) content rather than cytoplasmic Ca(2+) concentration. PMID- 11390927 TI - Calmodulin and Excitation-Contraction Coupling. AB - Excitation-contraction coupling in cardiac and skeletal muscle involves the transverse-tubule voltage-dependent Ca(2+) channel and the sarcoplasmic reticulum Ca(2+) release channel. Both of these ion channels bind and are modulated by calmodulin in both its Ca(2+)-bound and Ca(2+)-free forms. Calmodulin is, therefore, potentially an important regulator of excitation-contraction coupling. Its precise role, however, has not yet been defined. PMID- 11390928 TI - GABA in the Mammalian Enteric Nervous System. AB - gamma-Aminobutyric acid (GABA) is a transmitter of enteric interneurons, targeting excitatory GABA(A) or inhibitory GABA(B) receptors that modulate motility and mucosal function. Enteric GABA may also subserve hormonal and paracrine signaling. Disruption in gastrointestinal function following perturbation of enteric GABA receptors presents potential new target sites for drug development. PMID- 11390930 TI - Inflammatory Cytokines in Nonpathological States. AB - During infection, inflammatory cytokines induce regulated changes in the host's internal milieu that create a hostile environment to an invading pathogen. Recent evidence indicates that these cytokines are constitutively produced, their production is increased by environmental stressors other than microbes, and they modulate "normal" physiological processes. PMID- 11390929 TI - A Novel Pacemaker Mechanism Drives Gastrointestinal Rhythmicity. AB - Electric pacemaker activity drives peristaltic and segmental contractions in the gastrointestinal tract. Interstitial cells of Cajal (ICC) are responsible for spontaneous pacemaker activity. ICC remain rhythmic in culture and generate voltage-independent inward currents via a nonselective cation conductance. Ca(2+) release from endoplasmic reticulum and uptake by mitochondria initiates pacemaker currents. This novel mechanism provides the basis for electric rhythmicity in gastrointestinal muscles. PMID- 11390931 TI - Myocardial Reperfusion Injury: Insights Gained from Gene-Targeted Mice. AB - Myocardial ischemia-reperfusion injury involves activation of multiple cell types, including leukocytes and endothelial cells. The pursuant inflammation involves diminished nitric oxide production, influx of neutrophils, and myocardial cell injury. Gene-targeted animals provide important clues about the progression of this inflammatory cascade. PMID- 11390932 TI - Cation-Chloride Cotransporters in Neuronal Communication. AB - Two isoforms of the cation-Cl(-) cotransporter family are expressed in neurons and modulate neurotransmission. NKCC1, a Na(+)-K(+)-2Cl(-) cotransporter, by raising internal Cl(-), is responsible for excitatory GABAergic activity in immature brain and in adult sensory neurons. KCC2, a neuronal-specific isoform of the K(+)-Cl(-) cotransporter, by lowering internal Cl(-), is critical for inhibitory GABA responses in mature central nervous system neurons. PMID- 11390933 TI - Uncoupling Proteins: Do They Have a Role in Body Weight Regulation? AB - Several members of the mitochondrial carrier protein family are classified as uncoupling proteins. In contrast to the uncoupling protein specific to brown adipose tissue (UCP1), the physiological role of skeletal muscle uncoupling proteins (UCP2 and UCP3) in weight regulation seems more closely associated with the regulation of lipids as fuel substrate than as mediators of adaptive thermogenesis. PMID- 11390935 TI - Toward an Integrative Concept of Control of Total Body Sodium. AB - Total body sodium (TBSodium) is a major determinant of body water and arterial pressure. Several observations, in particular that of a "sodium memory," indicate that TBSodium is a controlled variable. Various regulatory elements are involved, e.g., the renin-angiotensin-aldosterone system, atrial receptors, and renal arterial pressure. Balance studies in dogs provide new insights into their contributions to TBSodium control. PMID- 11390936 TI - Propagation of Cell Death During Myocardial Reperfusion. AB - During myocardial reperfusion, increased cytosolic Ca(2+) concentration may cause hypercontracture and cell death. Hypercontracture can propagate to adjacent cells by a gap junction-dependent mechanism. This propagation explains infarct geometry and increases the final extent of necrosis. Its prevention may represent a new therapeutic strategy for treating patients with myocardial infarction. PMID- 11390937 TI - UTP as an extracellular signaling molecule. AB - In addition to their central role in many biochemical processes, uridine nucleotides are important extracellular signaling molecules that regulate a broad spectrum of cell functions via activation of P2Y2, P2Y4, and P2Y6 receptors. Cellular release of UTP provides a mechanism for autocrine control of calcium- or protein kinase C-dependent processes. PMID- 11390938 TI - Calcium, cross-bridges, and the Frank-Starling relationship. AB - The steep relationship between active force and length in cardiac muscle is based on a length dependence of myofilament Ca(2+) sensitivity. However, it is not muscle length but the lateral spacing between actin and myosin filaments that sets the level of Ca(2+) sensitivity, mainly through modulation of myosin mediated activation of the thin filament. PMID- 11390939 TI - Measurements of vesicle recycling in central neurons. AB - Neurotransmitter-containing vesicles in presynaptic nerve terminals are essential for synaptic transmission. The vesicles undergo a cycle that leads to transmitter release by exocytosis followed by endocytosis and refilling of the vesicles. Here we discuss new optical methods that have helped researchers study this cycle at functional and molecular levels, which is essential for our understanding of the regulation of synaptic transmission. PMID- 11390940 TI - Neutrophil diapedesis: paracellular or transcellular? AB - To reach an inflammatory site in the interstitium, circulating neutrophils (PMN) must first traverse the endothelial barrier. Whether PMN emigrate between endothelial cells (paracellular pathway) or through the endothelial cells proper (transcellular pathway) is controversial. Herein, we present anatomic, functional, and teleological arguments that support both points of view. An attempt is also made to reconcile this apparent controversy. PMID- 11390941 TI - Implications of mouse genotype for phenotype. AB - Genetic engineering of inbred mice offers the capability to separate interactions of multiple genes that control host resistance to pathogens. An understanding of the negative impact that genetic variability of mouse substrains can have on data is necessary for the design of experiments to dissect out complicated gene interactions. PMID- 11390942 TI - Gastroduodenal mucosal alkaline secretion and mucosal protection. AB - The gastroduodenal mucosa is a dynamic barrier restricting entry of gastric acid and other potentially hostile luminal contents. Mucosal HCO3(-) is a key element in preventing epithelial damage, and knowledge about HCO3(-) transport processes, including the role of the cystic fibrosis transmembrane conductance regulator channel, and their neurohumoral control are in rapid progress. PMID- 11390943 TI - Ion channels and transporters on the move. AB - Cell migration plays a crucial role in a variety of (patho)physiological processes such as immune defense, wound healing, and formation of tumor metastases. Detailed models have been developed to describe cytoskeletal mechanisms of migration. However, evidence is accumulating that the activity of ion channels and transporters is also required for optimal cell locomotion. PMID- 11390944 TI - Neurogenic inflammation in human and rodent skin. AB - The combination of vasodilation and protein extravasation following activation of nociceptors has been termed "neurogenic inflammation." In contrast to rodents, no neurogenic protein extravasation can be elicited in healthy human skin. Dermal microdialysis has considerably increased our knowledge about neurogenic inflammation in human skin, including the involvement of mast cells. PMID- 11390946 TI - Physiological properties of blood substitutes. AB - Blood substitutes (modified hemoglobin solutions, perfluorocarbon emulsions) serve as artificial oxygen carriers and are alternatives to blood transfusions. Hemoglobin solutions mimic the sigmoidal oxygen dissociation curve of natural blood. Perfluorocarbon emulsions exhibit a linear relation between PO2 and dissolved oxygen. The most advanced substances may enter medicine in few years. PMID- 11390947 TI - Apoptosis in the heart: about programmed cell death and survival. AB - Substantial evidence has accumulated that apoptosis, sometimes called "programmed cell death," is important in several cardiac diseases. Although most researchers focus on apoptosis in the hope that by understanding its mechanisms one can block this form of cell death, little attention has been given to programmed cell survival. PMID- 11390948 TI - Skeletal and cardiac muscle contractile activation: tropomyosin "rocks and rolls". AB - Changes in thin filament structure induced by Ca(2+) binding to troponin and subsequent strong cross-bridge binding regulate additional strong cross-bridge attachment, force development, and dependence of force on sarcomere length in skeletal and cardiac muscle. Variations in activation properties account for functional differences between these muscle types. PMID- 11390949 TI - Traffic control: Rab GTPases and the regulation of interorganellar transport. AB - Membrane proteins, membrane lipids, and luminal contents are exchanged among the intracellular compartments of eukaryotic cells by vesicular transport. This process must be highly ordered to maintain cellular architecture in the face of rapid membrane turnover. The Ras-related Rab GTPases play multiple roles in regulating this traffic. PMID- 11390950 TI - Smooth muscle excitation-contraction coupling: a role for caveolae and caveolins? AB - Agonist stimulation of smooth muscle contractility involves integration of many signal-transducing events from the plasma membrane to myofilaments in the cytoplasm. Recent evidence suggests an important role for membranous invaginations termed caveolae, and their integral protein components caveolins, in the coordination of extracellular contractile stimuli and intracellular effectors in smooth muscle. PMID- 11390951 TI - Development of lung edema: interstitial fluid dynamics and molecular structure. AB - Pulmonary interstitium is maintained dehydrated at subatmospheric pressure (-10 cmH(2)O) through low capillary permeability, low tissue compliance, and an efficient lymphatic drainage. Enzymatic degradation of proteoglycans disrupts the endothelial basal membrane and the matrix structure, triggering the development of pulmonary edema. PMID- 11390952 TI - Brain glucose transporters: relationship to local energy demand. AB - Glucose, the major fuel in the brain, is transported across the cell membranes by facilitated diffusion mediated by glucose transporter proteins. Essentially two types of glucose transporters are localized in the membranes of brain endothelial cells, astrocytes, and neurons. Their densities are well adjusted to changes in local energy demand. PMID- 11390953 TI - Leukocyte recruitment in the microcirculation: the rolling paradigm revisited. AB - Intravital microscopy has done much to elucidate the cascade of events involved in the recruitment of leukocytes to sites of inflammation. Here we review the physiological relevance of leukocyte rolling and some of the important subtleties of this process, highlighting limitations in our knowledge and directions for future investigation. PMID- 11390954 TI - Physiological insights from genetic manipulation of the renin-angiotensin system. AB - The renin-angiotensin system is one of the most widely studied endocrine systems. It has an important role in the regulation of normal homeostasis, and disturbances in this system may be important in numerous pathological states. This review will focus on the major insights and important questions raised from gene targeting of this system. PMID- 11390955 TI - Vascular endothelium: checkpoint for inflammation and immunity. AB - Vascular endothelial cells play a threefold role in the interaction with leukocytes. First, they are gatekeepers in leukocyte recruitment to inflammatory foci and lymphocyte homing to secondary lymphoid organs. Second, they modulate leukocyte activation. Finally, they are targets of leukocyte-derived molecules, resulting either in endothelial cell activation or death. PMID- 11390956 TI - An autocrine/paracrine mechanism triggered by myocardial stretch induces changes in contractility. AB - An autocrine/paracrine mechanism is triggered by stretching the myocardium. This mechanism involves release of angiotensin II, release/increased formation of endothelin, activation of the Na(+)/H(+) exchanger, increase in intracellular Na(+), and the increase in the Ca(2+) transient that underlies the slow force response to stretch. The autocrine/paracrine mechanism could explain how changes in afterload alter cardiac contractility. PMID- 11390957 TI - Asymmetry of the neuroendocrine system. AB - There is information on the lateralization of hypothalamic, limbic, and other brain structures involved in the control of the endocrine glands. Sided differences between paired glands, including their peripheral innervation, and relevant clinical observations on asymmetry are also known. Data suggest predominance of the right half of brain structures in controlling gonadal function. PMID- 11390958 TI - Excitatory synaptic transmission in neonatal dorsal horn: NMDA and ATP receptors. AB - Postnatal development sees a strong synaptogenesis in rat superficial dorsal horn. My studies show that synapses mediated by two excitatory neurotransmitters, glutamate and ATP, are functional since the very first postnatal days. Using an electrophysiological approach, the functional properties of two receptors activated by these neurotransmitters, glutamatergic NMDA and ATP ionotropic receptors, are described. PMID- 11390960 TI - Corneal collagen fibril structure in three dimensions: Structural insights into fibril assembly, mechanical properties, and tissue organization. AB - The ability of the cornea to transmit light while being mechanically resilient is directly attributable to the formation of an extracellular matrix containing orthogonal sheets of collagen fibrils. The detailed structure of the fibrils and how this structure underpins the mechanical properties and organization of the cornea is understood poorly. In this study, we used automated electron tomography to study the three-dimensional organization of molecules in corneal collagen fibrils. The reconstructions show that the collagen molecules in the 36-nm diameter collagen fibrils are organized into microfibrils (approximately 4-nm diameter) that are tilted by approximately 15 degrees to the fibril long axis in a right-handed helix. An unexpected finding was that the microfibrils exhibit a constant-tilt angle independent of radial position within the fibril. This feature suggests that microfibrils in concentric layers are not always parallel to each other and cannot retain the same neighbors between layers. Analysis of the lateral structure shows that the microfibrils exhibit regions of order and disorder within the 67-nm axial repeat of collagen fibrils. Furthermore, the microfibrils are ordered at three specific regions of the axial repeat of collagen fibrils that correspond to the N- and C-telopeptides and the d-band of the gap zone. The reconstructions also show macromolecules binding to the fibril surface at sites that correspond precisely to where the microfibrils are most orderly. PMID- 11390961 TI - On the origin of and phylogenetic relationships among living amphibians. AB - The phylogenetic relationships among the three orders of modern amphibians (Caudata, Gymnophiona, and Anura) have been estimated based on both morphological and molecular evidence. Most morphological and paleontological studies of living and fossil amphibians support the hypothesis that salamanders and frogs are sister lineages (the Batrachia hypothesis) and that caecilians are more distantly related. Previous interpretations of molecular data based on nuclear and mitochondrial rRNA sequences suggested that salamanders and caecilians are sister groups to the exclusion of frogs. In an attempt to resolve this apparent conflict, the complete mitochondrial genomes of a salamander (Mertensiella luschani) and a caecilian (Typhlonectes natans) were determined (16,656 and 17,005 bp, respectively) and compared with previously published sequences from a frog (Xenopus laevis) and several other groups of vertebrates. Phylogenetic analyses of the mitochondrial data supported with high bootstrap values the monophyly of living amphibians with respect to other living groups of tetrapods, and a sister group relationship of salamanders and frogs. The lack of phylogenetically informative sites in the previous rRNA data sets (because of its shorter size and higher among-site rate variation) likely explains the discrepancy between our results and those based on previous molecular data. Strong support of the Batrachia hypothesis from both molecule- and morphology based studies provides a robust phylogenetic framework that will be helpful to comparative studies among the three living orders of amphibians and will permit better understanding of the considerably divergent vertebral, brain, and digit developmental patterns found in frogs and salamanders. PMID- 11390962 TI - Tracking wakes: the nocturnal predatory strategy of piscivorous catfish. AB - Swimming fish leave wakes containing hydrodynamic and chemical traces. These traces mark their swim paths and could guide predators. We now show that nocturnal European catfish (Silurus glanis) locate a piscine prey (guppy, Poecilia reticulata) by accurately tracking its three-dimensional swim path before an attack in the absence of visible light. Wakes that were up to 10 s old were followed over distances up to 55 prey-body lengths in our setup. These results demonstrate that prey wakes remain sufficiently identifiable to guide predators, and to extend considerably the area in which prey is detectable. Moreover, wakes elicit rear attacks, which may be more difficult to detect by prey. Wake tracking may be a common strategy among aquatic predators. PMID- 11390963 TI - Adenylate kinase phosphotransfer communicates cellular energetic signals to ATP sensitive potassium channels. AB - Transduction of energetic signals into membrane electrical events governs vital cellular functions, ranging from hormone secretion and cytoprotection to appetite control and hair growth. Central to the regulation of such diverse cellular processes are the metabolism sensing ATP-sensitive K+ (K(ATP)) channels. However, the mechanism that communicates metabolic signals and integrates cellular energetics with K(ATP) channel-dependent membrane excitability remains elusive. Here, we identify that the response of K(ATP) channels to metabolic challenge is regulated by adenylate kinase phosphotransfer. Adenylate kinase associates with the K(ATP) channel complex, anchoring cellular phosphotransfer networks and facilitating delivery of mitochondrial signals to the membrane environment. Deletion of the adenylate kinase gene compromised nucleotide exchange at the channel site and impeded communication between mitochondria and K(ATP) channels, rendering cellular metabolic sensing defective. Assigning a signal processing role to adenylate kinase identifies a phosphorelay mechanism essential for efficient coupling of cellular energetics with K(ATP) channels and associated functions. PMID- 11390965 TI - Aeolian dust in Colorado Plateau soils: nutrient inputs and recent change in source. AB - Aeolian dust (windblown silt and clay) is an important component in arid-land ecosystems because it may contribute to soil formation and furnish essential nutrients. Few geologic surfaces, however, have been characterized with respect to dust-accumulation history and resultant nutrient enrichment. We have developed a combination of methods to identify the presence of aeolian dust in arid regions and to evaluate the roles of this dust in ecosystem processes. Unconsolidated sandy sediment on isolated surfaces in the Canyonlands region of the Colorado Plateau differs greatly in mineralogical and chemical composition from associated bedrock, mainly aeolian sandstone. Detrital magnetite in the surficial deposits produces moderately high values of magnetic susceptibility, but magnetite is absent in nearby bedrock. A component of the surficial deposits must be aeolian to account for the abundance of magnetite, which formed originally in far-distant igneous rocks. Particle-size analysis suggests that the aeolian dust component is typically as much as 20-30%. Dust inputs have enriched the sediments in many elements, including P, Mg, Na, K, and Mo, as well as Ca, at sites where bedrock lacks calcite cement. Soil-surface biologic crusts are effective dust traps that apparently record a change in dust sources over the past several decades. Some of the recently fallen dust may result from human disturbance of land surfaces that are far from the Canyonlands, such as the Mojave Desert. Some land-use practices in the study area have the potential to deplete soil fertility by means of wind erosion removal of aeolian silt. PMID- 11390964 TI - Kinetics and mechanism of DNA uptake into the cell nucleus. AB - Gene transfer to eukaryotic cells requires the uptake of exogenous DNA into the cell nucleus. Except during mitosis, molecular access to the nuclear interior is limited to passage through the nuclear pores. Here we demonstrate the nuclear uptake of extended linear DNA molecules by a combination of fluorescence microscopy and single-molecule manipulation techniques, using the latter to follow uptake kinetics of individual molecules in real time. The assays were carried out on nuclei reconstituted in vitro from extracts of Xenopus eggs, which provide both a complete complement of biochemical factors involved in nuclear protein import, and unobstructed access to the nuclear pores. We find that uptake of DNA is independent of ATP or GTP hydrolysis, but is blocked by wheat germ agglutinin. The kinetics are much slower than would be expected from hydrodynamic considerations. A fit of the data to a simple model suggests femto-Newton forces and a large friction relevant to the uptake process. PMID- 11390966 TI - Tissue-specific overexpression of lipoprotein lipase causes tissue-specific insulin resistance. AB - Insulin resistance in skeletal muscle and liver may play a primary role in the development of type 2 diabetes mellitus, and the mechanism by which insulin resistance occurs may be related to alterations in fat metabolism. Transgenic mice with muscle- and liver-specific overexpression of lipoprotein lipase were studied during a 2-h hyperinsulinemic-euglycemic clamp to determine the effect of tissue-specific increase in fat on insulin action and signaling. Muscle lipoprotein lipase mice had a 3-fold increase in muscle triglyceride content and were insulin resistant because of decreases in insulin-stimulated glucose uptake in skeletal muscle and insulin activation of insulin receptor substrate-1 associated phosphatidylinositol 3-kinase activity. In contrast, liver-lipoprotein lipase mice had a 2-fold increase in liver triglyceride content and were insulin resistant because of impaired ability of insulin to suppress endogenous glucose production associated with defects in insulin activation of insulin receptor substrate-2-associated phosphatidylinositol 3-kinase activity. These defects in insulin action and signaling were associated with increases in intracellular fatty acid-derived metabolites (i.e., diacylglycerol, fatty acyl CoA, ceramides). Our findings suggest a direct and causative relationship between the accumulation of intracellular fatty acid-derived metabolites and insulin resistance mediated via alterations in the insulin signaling pathway, independent of circulating adipocyte-derived hormones. PMID- 11390968 TI - Thirty years on the "broad spectrum revolution" and paleolithic demography. PMID- 11390967 TI - Kinetic proofreading models for cell signaling predict ways to escape kinetic proofreading. AB - In the context of cell signaling, kinetic proofreading was introduced to explain how cells can discriminate among ligands based on a kinetic parameter, the ligand receptor dissociation rate constant. In the kinetic proofreading model of cell signaling, responses occur only when a bound receptor undergoes a complete series of modifications. If the ligand dissociates prematurely, the receptor returns to its basal state and signaling is frustrated. We extend the model to deal with systems where aggregation of receptors is essential to signal transduction, and present a version of the model for systems where signaling depends on an extrinsic kinase. We also investigate the kinetics of signaling molecules, "messengers," that are generated by aggregated receptors but do not remain associated with the receptor complex. We show that the extended model predicts modes of signaling that exhibit kinetic discrimination for some range of parameters but for other parameter values show little or no discrimination and thus escape kinetic proofreading. We compare model predictions with experimental data. PMID- 11390969 TI - A crystallographic map of the transition from B-DNA to A-DNA. AB - The transition between B- and A-DNA was first observed nearly 50 years ago. We have now mapped this transformation through a set of single-crystal structures of the sequence d(GGCGCC)2, with various intermediates being trapped by methylating or brominating the cytosine bases. The resulting pathway progresses through 13 conformational steps, with a composite structure that pairs A-nucleotides with complementary B-nucleotides serving as a distinct transition intermediate. The details of each step in the conversion of B- to A-DNA are thus revealed at the atomic level, placing intermediates for this and other sequences in the context of a common pathway. PMID- 11390970 TI - Global modulation of cellular transcription by human cytomegalovirus is initiated by viral glycoprotein B. AB - Human cytomegalovirus (HCMV) infection alters the expression of many cellular genes, including IFN-stimulated genes (ISGs) [Zhu, H., Cong, J.-P., Mamtora, G., Gingeras, T. & Shenk, T. (1998) Proc. Natl. Acad. Sci. USA 95, 14470-14475]. By using high-density cDNA microarrays, we show that the HCMV-regulated gene expression profile in fibroblasts does not differ substantially from the response generated by IFN. Furthermore, we identified the specific viral component triggering this response as the envelope glycoprotein B (gB). Cells treated with gB, but not other herpesviral glycoproteins, exhibited the same transcriptional profile as HCMV-infected cells. Thus, the interaction of gB with its as yet unidentified cellular receptor is the principal mechanism by which HCMV alters cellular gene expression early during infection. These findings highlight a pioneering paradigm for the consequences of virus-receptor interactions. PMID- 11390971 TI - Effects of anesthesia on functional activation of cerebral blood flow and metabolism. AB - Functional brain mapping based on changes in local cerebral blood flow (lCBF) or glucose utilization (lCMR(glc)) induced by functional activation is generally carried out in animals under anesthesia, usually alpha-chloralose because of its lesser effects on cardiovascular, respiratory, and reflex functions. Results of studies on the role of nitric oxide (NO) in the mechanism of functional activation of lCBF have differed in unanesthetized and anesthetized animals. NO synthase inhibition markedly attenuates or eliminates the lCBF responses in anesthetized animals but not in unanesthetized animals. The present study examines in conscious rats and rats anesthetized with alpha-chloralose the effects of vibrissal stimulation on lCMR(glc) and lCBF in the whisker-to-barrel cortex pathway and on the effects of NO synthase inhibition with N(G)-nitro-L arginine methyl ester (L-NAME) on the magnitude of the responses. Anesthesia markedly reduced the lCBF and lCMR(glc) responses in the ventral posteromedial thalamic nucleus and barrel cortex but not in the spinal and principal trigeminal nuclei. L-NAME did not alter the lCBF responses in any of the structures of the pathway in the unanesthetized rats and also not in the trigeminal nuclei of the anesthetized rats. In the thalamus and sensory cortex of the anesthetized rats, where the lCBF responses to stimulation had already been drastically diminished by the anesthesia, L-NAME treatment resulted in loss of statistically significant activation of lCBF by vibrissal stimulation. These results indicate that NO does not mediate functional activation of lCBF under physiological conditions. PMID- 11390972 TI - Identification and characterization of a lysosomal transporter for small neutral amino acids. AB - In eukaryotic cells, lysosomes represent a major site for macromolecule degradation. Hydrolysis products are eventually exported from this acidic organelle into the cytosol through specific transporters. Impairment of this process at either the hydrolysis or the efflux step is responsible of several lysosomal storage diseases. However, most lysosomal transporters, although biochemically characterized, remain unknown at the molecular level. In this study, we report the molecular and functional characterization of a lysosomal amino acid transporter (LYAAT-1), remotely related to a family of H+-coupled plasma membrane and synaptic vesicle amino acid transporters. LYAAT-1 is expressed in most rat tissues, with highest levels in the brain where it is present in neurons. Upon overexpression in COS-7 cells, the recombinant protein mediates the accumulation of neutral amino acids, such as gamma-aminobutyric acid, l-alanine, and l-proline, through an H+/amino acid symport. Confocal microscopy on brain sections revealed that this transporter colocalizes with cathepsin D, an established lysosomal marker. LYAAT-1 thus appears as a lysosomal transporter that actively exports neutral amino acids from lysosomes by chemiosmotic coupling to the H+-ATPase of these organelles. Homology searching in eukaryotic genomes suggests that LYAAT-1 defines a subgroup of lysosomal transporters in the amino acid/auxin permease family. PMID- 11390973 TI - Structural basis for fibroblast growth factor receptor 2 activation in Apert syndrome. AB - Apert syndrome (AS) is characterized by craniosynostosis (premature fusion of cranial sutures) and severe syndactyly of the hands and feet. Two activating mutations, Ser-252 --> Trp and Pro-253 --> Arg, in fibroblast growth factor receptor 2 (FGFR2) account for nearly all known cases of AS. To elucidate the mechanism by which these substitutions cause AS, we determined the crystal structures of these two FGFR2 mutants in complex with fibroblast growth factor 2 (FGF2). These structures demonstrate that both mutations introduce additional interactions between FGFR2 and FGF2, thereby augmenting FGFR2-FGF2 affinity. Moreover, based on these structures and sequence alignment of the FGF family, we propose that the Pro-253 --> Arg mutation will indiscriminately increase the affinity of FGFR2 toward any FGF. In contrast, the Ser-252 --> Trp mutation will selectively enhance the affinity of FGFR2 toward a limited subset of FGFs. These predictions are consistent with previous biochemical data describing the effects of AS mutations on FGF binding. Alterations in FGFR2 ligand affinity and specificity may allow inappropriate autocrine or paracrine activation of FGFR2. Furthermore, the distinct gain-of-function interactions observed in each crystal structure provide a model to explain the phenotypic variability among AS patients. PMID- 11390974 TI - Transcription factor RF2a alters expression of the rice tungro bacilliform virus promoter in transgenic tobacco plants. AB - The promoter from rice tungro bacilliform badnavirus (RTBV) is expressed only in phloem tissues in transgenic rice plants. RF2a, a b-Zip protein from rice, is known to bind to the Box II cis element near the TATA box of the promoter. Here, we report that the full-length RTBV promoter and a truncated fragment E of the promoter, comprising nucleotides -164 to +45, result in phloem-specific expression of beta-glucuronidase (GUS) reporter genes in transgenic tobacco plants. When a fusion gene comprising the cauliflower mosaic virus 35S promoter and RF2a cDNA was coexpressed with the GUS reporter genes, GUS activity was increased by 2-20-fold. The increase in GUS activity was positively correlated with the amount of RF2a, and the expression pattern of the RTBV promoter was altered from phloem-specific to constitutive. Constitutive expression of RF2a did not induce morphological changes in the transgenic plants. In contrast, constitutive overexpression of the b-ZIP domain of RF2a had a strong effect on the development of transgenic plants. These studies suggest that expression of the b-Zip domain can interfere with the function of homologues of RF2a that regulate development of tobacco plants. PMID- 11390975 TI - Heteromeric association creates a P2Y-like adenosine receptor. AB - Adenosine and its endogenous precursor ATP are main components of the purinergic system that modulates cellular and tissue functions via specific adenosine and ATP receptors (P1 and P2 receptors), respectively. Although adenosine inhibits excitability and ATP functions as an excitatory transmitter in the central nervous system, little is known about the ability of P1 and P2 receptors to form new functional structures such as a heteromer to control the complex purinergic cascade. Here we have shown that G(i/o) protein-coupled A1 adenosine receptor (A1R) and Gq protein-coupled P2Y1 receptor (P2Y1R) coimmunoprecipitate in cotransfected HEK293T cells, suggesting the oligomeric association between distinct G protein-coupled P1 and P2 receptors. A1R and P2Y2 receptor, but not A1R and dopamine D2 receptor, also were found to coimmunoprecipitate in cotransfected cells. A1R agonist and antagonist binding to cell membranes were reduced by coexpression of A1R and P2Y1R, whereas a potent P2Y1R agonist adenosine 5'-O-(2-thiotriphosphate) (ADPbetaS) revealed a significant potency to A1R binding only in the cotransfected cell membranes. Moreover, the A1R/P2Y1R coexpressed cells showed an ADPbetaS-dependent reduction of forskolin-evoked cAMP accumulation that was sensitive to pertussis toxin and A1R antagonist, indicating that ADPbetaS binds A1R and inhibits adenylyl cyclase activity via G(i/o) proteins. Also, a high degree of A1R and P2Y1R colocalization was demonstrated in cotransfected cells by double immunofluorescence experiments with confocal laser microscopy. These results suggest that oligomeric association of A1R with P2Y1R generates A1R with P2Y1R-like agonistic pharmacology and provides a molecular mechanism for an increased diversity of purine signaling. PMID- 11390976 TI - Ornaments of the earliest Upper Paleolithic: new insights from the Levant. AB - Two sites located on the northern Levantine coast, Ucagizli Cave (Turkey) and Ksar 'Akil (Lebanon) have yielded numerous marine shell beads in association with early Upper Paleolithic stone tools. Accelerator mass spectrometry (AMS) radiocarbon dates indicate ages between 39,000 and 41,000 radiocarbon years (roughly 41,000-43,000 calendar years) for the oldest ornament-bearing levels in Ucagizli Cave. Based on stratigraphic evidence, the earliest shell beads from Ksar 'Akil may be even older. These artifacts provide some of the earliest evidence for traditions of personal ornament manufacture by Upper Paleolithic humans in western Asia, comparable in age to similar objects from Eastern Europe and Africa. The new data show that the initial appearance of Upper Paleolithic ornament technologies was essentially simultaneous on three continents. The early appearance and proliferation of ornament technologies appears to have been contingent on variable demographic or social conditions. PMID- 11390977 TI - Thrombin induces the release of the Y-box protein dbpB from mRNA: a mechanism of transcriptional activation. AB - We have recently demonstrated that thrombin induces expression of the platelet derived growth factor B-chain gene in endothelial cells (EC) through activation of the Y-box binding protein DNA-binding protein B (dbpB). We now present evidence that dbpB is activated by a novel mechanism: proteolytic cleavage leading to release from mRNA, nuclear translocation, and induction of thrombin responsive genes. Cytosolic, full-length dbpB (50 kDa) was rapidly cleaved to a 30-kDa species upon thrombin stimulation of EC. This truncated, "active" dbpB exhibited nuclear localization and binding affinity for the thrombin response element sequence, which is distinct from the Y-box sequence. Oligo(dT) affinity chromatography revealed that cytosolic dbpB from control EC, but not active dbpB from thrombin-treated EC, was bound to mRNA. Latent dbpB immunoprecipitated from cytosolic extracts of control EC was activated by ribonuclease treatment. Furthermore, when EC cytosolic extracts were subjected to Nycodenz gradient centrifugation, latent dbpB fractionated with mRNA, whereas active dbpB fractionated with free proteins. The cytosolic retention domain of dbpB, which we localized to the region 247-267, was proteolytically cleaved during its activation. In contrast to full-length dbpB, truncated dbpB stimulated platelet derived growth factor B-chain and tissue factor promoter activity by over 5-fold when transiently cotransfected with reporter constructs. These results suggest a novel mode of transcription factor activation in which an agonist causes release from mRNA of a latent transcription factor leading to its transport to the nucleus and its regulation of target gene expression. PMID- 11390978 TI - Dopamine receptor regulating factor, DRRF: a zinc finger transcription factor. AB - Dopamine receptor genes are under complex transcription control, determining their unique regional distribution in the brain. We describe here a zinc finger type transcription factor, designated dopamine receptor regulating factor (DRRF), which binds to GC and GT boxes in the D1A and D2 dopamine receptor promoters and effectively displaces Sp1 and Sp3 from these sequences. Consequently, DRRF can modulate the activity of these dopamine receptor promoters. Highest DRRF mRNA levels are found in brain with a specific regional distribution including olfactory bulb and tubercle, nucleus accumbens, striatum, hippocampus, amygdala, and frontal cortex. Many of these brain regions also express abundant levels of various dopamine receptors. In vivo, DRRF itself can be regulated by manipulations of dopaminergic transmission. Mice treated with drugs that increase extracellular striatal dopamine levels (cocaine), block dopamine receptors (haloperidol), or destroy dopamine terminals (1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine) show significant alterations in DRRF mRNA. The latter observations provide a basis for dopamine receptor regulation after these manipulations. We conclude that DRRF is important for modulating dopaminergic transmission in the brain. PMID- 11390979 TI - A gene family required for human germ cell development evolved from an ancient meiotic gene conserved in metazoans. AB - The Deleted in AZoospermia (DAZ) genes encode potential RNA-binding proteins that are expressed exclusively in prenatal and postnatal germ cells and are strong candidates for human fertility factors. Here we report the identification of an additional member of the DAZ gene family, which we have called BOULE. With the identification of this gene, it is clear that the human DAZ gene family contains at least three members: DAZ, a Y-chromosome gene cluster that arose 30-40 million years ago and whose deletion is linked to infertility in men; DAZL, the "father" of DAZ, a gene that maps to human chromosome 3 and has homologs required for both female and male germ cell development in other organisms; and BOULE, a gene that we propose is the "grandfather" of DAZ and maps to human chromosome 2. Human and mouse BOULE resemble the invertebrate meiotic regulator Boule, the proposed ortholog of DAZ, in sequence and expression pattern and hence likely perform a similar meiotic function. In contrast, the previously identified human DAZ and DAZL are expressed much earlier than BOULE in prenatal germ stem cells and spermatogonia; DAZL also is expressed in female germ cells. These data suggest that homologs of the DAZ gene family can be grouped into two subfamilies (BOULE and DAZL) and that members of the DAZ family evolved from an ancestral meiotic regulator, Boule, to assume distinct, yet overlapping, functions in germ cell development. PMID- 11390980 TI - The role of haustoria in sugar supply during infection of broad bean by the rust fungus Uromyces fabae. AB - Biotrophic plant pathogenic fungi differentiate specialized infection structures within the living cells of their host plants. These haustoria have been linked to nutrient uptake ever since their discovery. We have for the first time to our knowledge shown that the flow of sugars from the host Vicia faba to the rust fungus Uromyces fabae seems to occur largely through the haustorial complex. One of the most abundantly expressed genes in rust haustoria, the expression of which is negligible in other fungal structures, codes for a hexose transporter. Functional expression of the gene termed HXT1 in Saccharomyces cerevisiae and Xenopus laevis oocytes assigned a substrate specificity for D-glucose and D fructose and indicated a proton symport mechanism. Abs against HXT1p exclusively labeled haustoria in immunofluorescence microscopy and the haustorial plasma membrane in electron microscopy. These results suggest that the fungus concentrates this transporter in haustoria to take advantage of a specialized compartment of the haustorial complex. The extrahaustorial matrix, delimited by the plasma membranes of both host and parasite, constitutes a newly formed apoplastic compartment with qualities distinct from those of the bulk apoplast. This organization might facilitate the competition of the parasite with natural sink organs of the host. PMID- 11390981 TI - Human tumor suppressor EXT gene family members EXTL1 and EXTL3 encode alpha 1,4- N-acetylglucosaminyltransferases that likely are involved in heparan sulfate/ heparin biosynthesis. AB - The tumor suppressors EXT1 and EXT2 are associated with hereditary multiple exostoses and encode bifunctional glycosyltransferases essential for chain polymerization of heparan sulfate (HS) and its analog, heparin (Hep). Three highly homologous EXT-like genes, EXTL1-EXTL3, have been cloned, and EXTL2 is an alpha1,4-GlcNAc transferase I, the key enzyme that initiates the HS/Hep synthesis. In the present study, truncated forms of EXTL1 and EXTL3, lacking the putative NH2-terminal transmembrane and cytoplasmic domains, were transiently expressed in COS-1 cells and found to harbor alpha-GlcNAc transferase activity. EXTL3 used not only N-acetylheparosan oligosaccharides that represent growing HS chains but also GlcAbeta1-3Galbeta1-O-C2H4NH-benzyloxycarbonyl (Cbz), a synthetic substrate for alpha-GlcNAc transferase I that determines and initiates HS/Hep synthesis. In contrast, EXTL1 used only the former acceptor. Neither EXTL1 nor EXTL3 showed any glucuronyltransferase activity as examined with N acetylheparosan oligosaccharides. Heparitinase I digestion of each transferase reaction product showed that GlcNAc had been transferred exclusively through an alpha1,4-configuration. Hence, EXTL3 most likely is involved in both chain initiation and elongation, whereas EXTL1 possibly is involved only in the chain elongation of HS and, maybe, Hep as well. Thus, their acceptor specificities of the five family members are overlapping but distinct from each other, except for EXT1 and EXT2 with the same specificity. It now has been clarified that all of the five cloned human EXT gene family proteins harbor glycosyltransferase activities, which probably contribute to the synthesis of HS and Hep. PMID- 11390982 TI - The transcriptional corepressor MITR is a signal-responsive inhibitor of myogenesis. AB - Activation of muscle-specific genes by members of the myocyte enhancer factor 2 (MEF2) and MyoD families of transcription factors is coupled to histone acetylation and is inhibited by class II histone deacetylases (HDACs) 4 and 5, which interact with MEF2. The ability of HDAC4 and -5 to inhibit MEF2 is blocked by phosphorylation of these HDACs at two conserved serine residues, which creates docking sites for the intracellular chaperone protein 14-3-3. When bound to 14-3 3, HDACs are released from MEF2 and transported to the cytoplasm, thereby allowing MEF2 to stimulate muscle-specific gene expression. MEF2-interacting transcription repressor (MITR) shares homology with the amino-terminal regions of HDAC4 and -5, but lacks an HDAC catalytic domain. Despite the absence of intrinsic HDAC activity, MITR acts as a potent inhibitor of MEF2-dependent transcription. Paradoxically, however, MITR has minimal inhibitory effects on the skeletal muscle differentiation program. We show that a substitution mutant of MITR containing alanine in place of two serine residues, Ser-218 and Ser-448, acts as a potent repressor of myogenesis. Our findings indicate that promyogenic signals antagonize the inhibitory action of MITR by targeting these serines for phosphorylation. Phosphorylation of Ser-218 and Ser-448 stimulates binding of 14 3-3 to MITR, disrupts MEF2:MITR interactions, and alters the nuclear distribution of MITR. These results reveal a role for MITR as a signal-dependent regulator of muscle differentiation. PMID- 11390983 TI - How the immune system works to protect the host from infection: a personal view. PMID- 11390984 TI - The candidate tumor suppressor gene, RASSF1A, from human chromosome 3p21.3 is involved in kidney tumorigenesis. AB - Clear cell-type renal cell carcinomas (clear RCC) are characterized almost universally by loss of heterozygosity on chromosome 3p, which usually involves any combination of three regions: 3p25-p26 (harboring the VHL gene), 3p12-p14.2 (containing the FHIT gene), and 3p21-p22, implying inactivation of the resident tumor-suppressor genes (TSGs). For the 3p21-p22 region, the affected TSGs remain, at present, unknown. Recently, the RAS association family 1 gene (isoform RASSF1A), located at 3p21.3, has been identified as a candidate lung and breast TSG. In this report, we demonstrate aberrant silencing by hypermethylation of RASSF1A in both VHL-caused clear RCC tumors and clear RCC without VHL inactivation. We found hypermethylation of RASSF1A's GC-rich putative promoter region in most of analyzed samples, including 39 of 43 primary tumors (91%). The promoter was methylated partially or completely in all 18 RCC cell lines analyzed. Methylation of the GC-rich putative RASSF1A promoter region and loss of transcription of the corresponding mRNA were related causally. RASSF1A expression was reactivated after treatment with 5-aza-2'-deoxycytidine. Forced expression of RASSF1A transcripts in KRC/Y, a renal carcinoma cell line containing a normal and expressed VHL gene, suppressed growth on plastic dishes and anchorage-independent colony formation in soft agar. Mutant RASSF1A had reduced growth suppression activity significantly. These data suggest that RASSF1A is the candidate renal TSG gene for the 3p21.3 region. PMID- 11390985 TI - Endogenous Msx1 antisense transcript: in vivo and in vitro evidences, structure, and potential involvement in skeleton development in mammals. AB - Msx1 is a key factor for the development of tooth and craniofacial skeleton and has been proposed to play a pivotal role in terminal cell differentiation. In this paper, we demonstrated the presence of an endogenous Msx1 antisense RNA (Msx1-AS RNA) in mice, rats, and humans. In situ analysis revealed that this RNA is expressed only in differentiated dental and bone cells with an inverse correlation with Msx1 protein. These in vivo data and overexpression of Msx1 sense and AS RNA in an odontoblastic cell line (MO6-G3) showed that the balance between the levels of the two Msx1 RNAs is related to the expression of Msx1 protein. To analyze the impact of this balance in the Msx-Dlx homeoprotein pathway, we analyzed the effect of Msx1, Msx2, and Dlx5 overexpression on proteins involved in skeletal differentiation. We showed that the Msx1-AS RNA is involved in crosstalk between the Msx-Dlx pathways because its expression was abolished by Dlx5. Msx1 was shown to down-regulate a master gene of skeletal cells differentiation, Cbfa1. All these data strongly suggest that the ratio between Msx1 sense and antisense RNAs is a very important factor in the control of skeletal terminal differentiation. Finally, the initiation site for Msx1-AS RNA transcription was located by primer extension in both mouse and human in an identical region, including a consensus TATA box, suggesting an evolutionary conservation of the AS RNA-mediated regulation of Msx1 gene expression. PMID- 11390986 TI - Clonable progenitors committed to the T lymphocyte lineage in the mouse bone marrow; use of an extrathymic pathway. AB - We searched for clonable committed T cell progenitors in the adult mouse bone marrow and isolated rare (approximately 0.05%) cells with the Thy-1hiCD2 CD16+CD44hiCD25-Lin- phenotype. In vivo experiments showed that these cells were progenitors committed only to reconstituting the T cell lineage of irradiated Ly5 congenic hosts. Reconstitution of the thymus was minimal compared with that of the bone marrow, spleen, and lymph nodes. At limiting dilutions, donor T cell reconstitution of the spleen frequently occurred without detectable donor cells in the thymus. Progenitors were capable of rapidly reconstituting athymic hosts. In conclusion, the clonable bone marrow progenitors were capable of T cell reconstitution predominantly by means of an extrathymic pathway. PMID- 11390987 TI - Molecular structures from femtosecond x-ray pulses. PMID- 11390988 TI - Interaction between vanilloid receptors and purinergic metabotropic receptors: pain perception and beyond. PMID- 11390989 TI - Mapping of complex regulatory elements by pufferfish/zebrafish transgenesis. PMID- 11390991 TI - The reduction in disability among the elderly. PMID- 11390990 TI - Mining copper transport genes. PMID- 11390992 TI - Total synthesis of cytochrome b562 by native chemical ligation using a removable auxiliary. AB - We have completed the total chemical synthesis of cytochrome b562 and an axial ligand analogue, [SeMet(7)]cyt b562, by thioester-mediated chemical ligation of unprotected peptide segments. A novel auxiliary-mediated native chemical ligation that enables peptide ligation to be applied to protein sequences lacking cysteine was used. A cleavable thiol-containing auxiliary group, 1-phenyl-2-mercaptoethyl, was added to the alpha-amino group of one peptide segment to facilitate amide bond-forming ligation. The amine-linked 1-phenyl-2-mercaptoethyl auxiliary was stable to anhydrous hydrogen fluoride used to cleave and deprotect peptides after solid-phase peptide synthesis. Following native chemical ligation with a thioester-containing segment, the auxiliary group was cleanly removed from the newly formed amide bond by treatment with anhydrous hydrogen fluoride, yielding a full-length unmodified polypeptide product. The resulting polypeptide was reconstituted with heme and folded to form the functional protein molecule. Synthetic wild-type cyt b562 exhibited spectroscopic and electrochemical properties identical to the recombinant protein, whereas the engineered [SeMet(7)]cyt b562 analogue protein was spectroscopically and functionally distinct, with a reduction potential shifted by approximately 45 mV. The use of the 1-phenyl-2-mercaptoethyl removable auxiliary reported here will greatly expand the applicability of total protein synthesis by native chemical ligation of unprotected peptide segments. PMID- 11390993 TI - An approach to three-dimensional structures of biomolecules by using single molecule diffraction images. AB - We describe an approach to the high-resolution three-dimensional structural determination of macromolecules that utilizes ultrashort, intense x-ray pulses to record diffraction data in combination with direct phase retrieval by the oversampling technique. It is shown that a simulated molecular diffraction pattern at 2.5-A resolution accumulated from multiple copies of single rubisco biomolecules, each generated by a femtosecond-level x-ray free electron laser pulse, can be successfully phased and transformed into an accurate electron density map comparable to that obtained by more conventional methods. The phase problem is solved by using an iterative algorithm with a random phase set as an initial input. The convergence speed of the algorithm is reasonably fast, typically around a few hundred iterations. This approach and phasing method do not require any ab initio information about the molecule, do not require an extended ordered lattice array, and can tolerate high noise and some missing intensity data at the center of the diffraction pattern. With the prospects of the x-ray free electron lasers, this approach could provide a major new opportunity for the high-resolution three-dimensional structure determination of single biomolecules. PMID- 11390994 TI - Stat1-independent regulation of gene expression in response to IFN-gamma. AB - Although Stat1 is essential for cells to respond fully to IFN-gamma, there is substantial evidence that, in the absence of Stat1, IFN-gamma can still regulate the expression of some genes, induce an antiviral state and affect cell growth. We have now identified many genes that are regulated by IFN-gamma in serum starved Stat1-null mouse fibroblasts. The proteins induced by IFN-gamma in Stat1 null cells can account for the substantial biological responses that remain. Some genes are induced in both wild-type and Stat1-null cells and thus are truly Stat1 independent. Others are subject to more complex regulation in response to IFN gamma, repressed by Stat1 in wild-type cells and activated in Stat1-null cells. Many genes induced by IFN-gamma in Stat1-null fibroblasts also are induced by platelet-derived growth factor in wild-type cells and thus are likely to be involved in cell proliferation. In mouse cells expressing the docking site mutant Y440F of human IFN-gamma receptor subunit 1, the mouse Stat1 is not phosphorylated in response to human IFN-gamma, but c-myc and c-jun are still induced, showing that the Stat1 docking site is not required for Stat1 independent signaling. PMID- 11390995 TI - Biologic consequences of Stat1-independent IFN signaling. AB - Although Stat1 is required for many IFN-dependent responses, recent work has shown that IFNgamma functions independently of Stat1 to affect the growth of tumor cells or immortalized fibroblasts. We now demonstrate that both IFNgamma and IFNalpha/beta regulate proliferative responses in cells of the mononuclear phagocyte lineage derived from Stat1-null mice. Using both representational difference analysis and gene arrays, we show that IFNgamma exerts its Stat1 independent actions on mononuclear phagocytes by regulating the expression of many genes. This result was confirmed by monitoring changes in expression and function of the corresponding gene products. Regulation of the expression of these genes requires the IFNgamma receptor and Jak1. The physiologic relevance of IFN-dependent, Stat1-independent signaling was demonstrated by monitoring antiviral responses in Stat1-null mice. Thus, the IFN receptors engage alternative Stat1-independent signaling pathways that have important physiological consequences. PMID- 11390996 TI - Genetic programs of epithelial cell plasticity directed by transforming growth factor-beta. AB - Epithelial-mesenchymal transitions (EMTs) are an essential manifestation of epithelial cell plasticity during morphogenesis, wound healing, and tumor progression. Transforming growth factor-beta (TGF-beta) modulates epithelial plasticity in these physiological contexts by inducing EMT. Here we report a transcriptome screen of genetic programs of TGF-beta-induced EMT in human keratinocytes and propose functional roles for extracellular response kinase (ERK) mitogen-activated protein kinase signaling in cell motility and disruption of adherens junctions. We used DNA arrays of 16,580 human cDNAs to identify 728 known genes regulated by TGF-beta within 4 hours after treatment. TGF-beta stimulated ERK signaling mediated regulation of 80 target genes not previously associated with this pathway. This subset is enriched for genes with defined roles in cell-matrix interactions, cell motility, and endocytosis. ERK independent genetic programs underlying the onset of EMT involve key pathways and regulators of epithelial dedifferentiation, undifferentiated transitional and mesenchymal progenitor phenotypes, and mediators of cytoskeletal reorganization. The gene expression profiling approach delineates complex context-dependent signaling pathways and transcriptional events that determine epithelial cell plasticity controlled by TGF-beta. Investigation of the identified pathways and genes will advance the understanding of molecular mechanisms that underlie tumor invasiveness and metastasis. PMID- 11390997 TI - Incipient speciation by sexual isolation in Drosophila: concurrent evolution at multiple loci. AB - Drosophila melanogaster from Zimbabwe and nearby regions shows strong but asymmetric sexual isolation from its cosmopolitan counterparts. By creating stable chromosome-substitution lines, earlier studies were able to show that the two major autosomes have very large effects on both male mating success and female mating preference. In this study, we genetically dissect this sexual isolation by recombination analysis between a whole-chromosome substitution line (which carries a Zimbabwe-derived third chromosome) and a strain with seven visible markers on that chromosome. Four loci are responsible for male mating success and three others are found to control female mating preference. Because male and female traits are not closely linked, their strong association among isofemale lines is most likely a reflection of sexual selection in nature. The results suggest that a large number of behavioral loci may evolve concurrently in the incipient stage of speciation before other aspects of reproductive isolation (such as hybrid sterility) have become evident. The results shed light on the population genetic processes underlying the formation of nascent species, as well as modes of speciation. PMID- 11390998 TI - A locus for female discrimination behavior causing sexual isolation in Drosophila. AB - The genetic basis of sexual isolation that contributes to speciation is one of the unsolved questions in evolutionary biology. Drosophila ananassae and Drosophila pallidosa are closely related, and postmating isolation has not developed between them. However, females of both species discriminate their mating partners, and this discrimination contributes to strong sexual isolation between them. By using surgical treatments, we demonstrate that male courtship songs play a dominant role in female mate discrimination. The absence of the song of D. pallidosa dramatically increased interspecies mating with D. ananassae females but reduced intraspecies mating with D. pallidosa females. Furthermore, genetic analysis and chromosomal introgression by repeated backcrosses to D. pallidosa males identified possible loci that control female discrimination in each species. These loci were mapped on distinct positions near the Delta locus on the middle of the left arm of the second chromosome. Because the mate discrimination we studied is well developed and is the only known mechanism that prevents gene flow between them, these loci may have played crucial roles in the evolution of reproductive isolation, and therefore, in the speciation process between these two species. PMID- 11390999 TI - Germ line deletion of the CD1 locus exacerbates diabetes in the NOD mouse. AB - Quantitative and qualitative defects in CD1-restricted natural killer T cells have been reported in several autoimmune-prone strains of mice, including the nonobese diabetic (NOD) mouse. These defects are believed to be associated with the emergence of spontaneous autoimmunity. Here we demonstrate that both CD1d null NOD and CD1d-null NOD/BDC2.5 T cell receptor transgenic mice have an accelerated onset and increased incidence of diabetes when compared with CD1d(+/ ) and CD1d(+/+) littermates. The acceleration of disease did not seem to result from changes in the T helper (Th)1/Th2 balance because lymphocytes purified from lymphoid organs and pancreatic islets of wild-type and CD1d-null mice secreted equivalent amounts of IFN-gamma and IL-4 after stimulation. In contrast, the pancreata of CD1d-null mice harbored significantly higher numbers of activated memory T cells expressing the chemokine receptor CCR4. Notably, the presence of these T cells was associated with immunohistochemical evidence of increased destructive insulitis. Thus, CD1d-restricted T cells are critically important for regulation of the spontaneous disease process in NOD mice. PMID- 11391000 TI - CD95/Fas induces cleavage of the GrpL/Gads adaptor and desensitization of antigen receptor signaling. AB - The balance between cell survival and cell death is critical for normal lymphoid development. This balance is maintained by signals through lymphocyte antigen receptors and death receptors such as CD95/Fas. In some cells, ligating the B cell antigen receptor can protect the cell from apoptosis induced by CD95. Here we report that ligation of CD95 inhibits antigen receptor-mediated signaling. Pretreating CD40-stimulated tonsillar B cells with anti-CD95 abolished B cell antigen receptor-mediated calcium mobilization. Furthermore, CD95 ligation led to the caspase-dependent inhibition of antigen receptor-induced calcium mobilization and to the activation of mitogen-activated protein kinase pathways in B and T cell lines. A target of CD95-mediated caspase 3-like activity early in the apoptotic process is the adaptor protein GrpL/Gads. GrpL constitutively interacts with SLP-76 via its C-terminal SH3 domain to regulate transcription factors such as NF-AT. Cleavage of GrpL removes the C-terminal SH3 domain so that it is no longer capable of recruiting SLP-76 to the membrane. Transfection of a truncated form of GrpL into Jurkat T cells blocked T cell antigen receptor-induced activation of NF-AT. These results suggest that CD95 signaling can desensitize antigen receptors, in part via cleavage of the GrpL adaptor. PMID- 11391001 TI - Antigen presentation subverted: Structure of the human cytomegalovirus protein US2 bound to the class I molecule HLA-A2. AB - Many persistent viruses have evolved the ability to subvert MHC class I antigen presentation. Indeed, human cytomegalovirus (HCMV) encodes at least four proteins that down-regulate cell-surface expression of class I. The HCMV unique short (US)2 glycoprotein binds newly synthesized class I molecules within the endoplasmic reticulum (ER) and subsequently targets them for proteasomal degradation. We report the crystal structure of US2 bound to the HLA-A2/Tax peptide complex. US2 associates with HLA-A2 at the junction of the peptide binding region and the alpha3 domain, a novel binding surface on class I that allows US2 to bind independently of peptide sequence. Mutation of class I heavy chains confirms the importance of this binding site in vivo. Available data on class I-ER chaperone interactions indicate that chaperones would not impede US2 binding. Unexpectedly, the US2 ER-luminal domain forms an Ig-like fold. A US2 structure-based sequence alignment reveals that seven HCMV proteins, at least three of which function in immune evasion, share the same fold as US2. The structure allows design of further experiments to determine how US2 targets class I molecules for degradation. PMID- 11391002 TI - Kinetics and thermodynamics of T cell receptor- autoantigen interactions in murine experimental autoimmune encephalomyelitis. AB - In the current study, cellular and molecular approaches have been used to analyze the biophysical nature of T cell receptor (TCR)-peptide MHC (pMHC) interactions for two autoreactive TCRs. These two TCRs recognize the N-terminal epitope of myelin basic protein (MBP1-11) bound to the MHC class II protein, I-A(u), and are associated with murine experimental autoimmune encephalomyelitis. Mice transgenic for the TCRs have been generated and characterized in other laboratories. These analyses indicate that the mice either develop encephalomyelitis spontaneously (172.10 TCR) or only if immunized with autoantigen in adjuvant (1934.4 TCR). Here, we show that the 172.10 TCR binds MBP1-11:I-A(u) with a 4-5-fold higher affinity than the 1934.4 TCR. Consistent with the higher affinity, 172.10 T hybridoma cells are significantly more responsive to autoantigen than 1934.4 cells. The interaction of the 172.10 TCR with cognate ligand is more entropically unfavorable than that of the 1934.4 TCR, indicating that the 172.10 TCR undergoes greater conformational rearrangements upon ligand binding. The studies therefore suggest a correlation between the strength and plasticity of a TCR-pMHC interaction and the frequency of spontaneous disease in the corresponding TCR transgenic mice. The comparative analysis of these two TCRs has implications for understanding autoreactive T cell recognition and activation. PMID- 11391003 TI - Dimerization and the effectiveness of ICAM-1 in mediating LFA-1-dependent adhesion. AB - Dimeric intercellular adhesion molecule-1 (ICAM-1) binds more efficiently to lymphocyte function-associated antigen-1 (LFA-1) than monomeric ICAM-1. However, it is unknown whether dimerization enhances binding simply by providing two ligand-binding sites and thereby increasing avidity, or whether it serves to generate a single "fully competent" LFA-1-binding surface. Domain 1 of ICAM-1 contains both the binding site for LFA-1, centered on residue E34, and a homodimerization interface. Whether the LFA-1-binding site extends across the homodimerization interface has not been tested. To address this question, we constructed four different heterodimeric soluble forms of ICAM-1 joined at the C terminus via an alpha-helical coiled coil (ACID-BASE). These heterodimeric ICAM-1 constructs include, (i) E34/E34 (two intact LFA-1-binding sites), (ii) E34/K34 (one disrupted LFA-1-binding site), (iii) E34/DeltaD1-2 (one deleted LFA-1 binding site), and (iv) K34/K34 (two disrupted LFA-1-binding sites). Cells bearing activated LFA-1 bound similarly to surfaces coated with either E34/K34 or E34/DeltaD1-2 and with an approximately 2-fold reduction in efficiency compared with E34/E34, suggesting that D1 dimerization, which is precluded in E34/DeltaD1 D2, is not necessary for optimal LFA-1 binding. Furthermore, BIAcore (BIAcore, Piscataway, NJ) affinity measurements revealed that soluble open LFA-1 I domain bound to immobilized soluble ICAM-1, E34/E34, E34/K34, and E34/DeltaD1-D2 with nearly identical affinities. These studies demonstrate that a single ICAM-1 monomer, not dimeric ICAM-1, represents the complete, "fully competent" LFA-1 binding surface. PMID- 11391004 TI - The copper transporter CTR1 provides an essential function in mammalian embryonic development. AB - Copper serves as an essential cofactor for a variety of proteins in all living organisms. Previously, we described a human gene (CTR1;SLC31A1) that encodes a high-affinity copper-uptake protein and hypothesized that this protein is required for copper delivery to mammalian cells. Here, we test this hypothesis by inactivating the Ctr1 gene in mice by targeted mutagenesis. We observe early embryonic lethality in homozygous mutant embryos and a deficiency in copper uptake in the brains of heterozygous animals. Ctr1(-/-) embryos can be recovered at E8.5 but are severely developmentally retarded and morphologically abnormal. Histological analysis reveals discontinuities and variable thickness in the basement membrane of the embryonic region and an imperfect Reichert's membrane, features that are likely due to lack of activity in the collagen cross-linking cupro-enzyme lysyl oxidase. A collapsed embryonic cavity, the absence of an allantois, retarded mesodermal migration, and increased cell death are also apparent. In the brains of heterozygous adult mice, which at 16 months are phenotypically normal, copper is reduced to approximately half compared with control littermates, implicating CTR1 as the required port for copper entry into at least this organ. A study of the spatial and temporal expression pattern of Ctr1 during mouse development and adulthood further shows that CTR1 is ubiquitously transcribed with highest expression observed in the specialized epithelia of the choroid plexus and renal tubules and in connective tissues of the eye, ovary, and testes. We conclude that CTR1 is the primary avenue for copper uptake in mammalian cells. PMID- 11391005 TI - Essential role for mammalian copper transporter Ctr1 in copper homeostasis and embryonic development. AB - The trace metal copper (Cu) plays an essential role in biology as a cofactor for many enzymes that include Cu, Zn superoxide dismutase, cytochrome oxidase, ceruloplasmin, lysyl oxidase, and dopamine beta-hydroxylase. Consequently, Cu transport at the cell surface and the delivery of Cu to intracellular compartments are critical events for a wide variety of biological processes. The components that orchestrate intracellular Cu trafficking and their roles in Cu homeostasis have been elucidated by the studies of model microorganisms and by the characterizations of molecular basis of Cu-related genetic diseases, including Menkes disease and Wilson disease. However, little is known about the mechanisms for Cu uptake at the plasma membrane and the consequences of defects in this process in mammals. Here, we show that the mouse Ctr1 gene encodes a component of the Cu transport machinery and that mice heterozygous for Ctr1 exhibit tissue-specific defects in copper accumulation and in the activities of copper-dependent enzymes. Mice completely deficient for Ctr1 exhibit profound growth and developmental defects and die in utero in mid-gestation. These results demonstrate a crucial role for Cu acquisition through the Ctr1 transporter for mammalian Cu homeostasis and embryonic development. PMID- 11391006 TI - The metallochaperone Atox1 plays a critical role in perinatal copper homeostasis. AB - Copper plays a fundamental role in the biochemistry of all aerobic organisms. The delivery of this metal to specific intracellular targets is mediated by metallochaperones. To elucidate the role of the metallochaperone Atox1, we analyzed mice with a disruption of the Atox1 locus. Atox1(-/-) mice failed to thrive immediately after birth, with 45% of pups dying before weaning. Surviving animals exhibited growth failure, skin laxity, hypopigmentation, and seizures because of perinatal copper deficiency. Maternal Atox1 deficiency markedly increased the severity of Atox1(-/-) phenotype, resulting in increased perinatal mortality as well as severe growth retardation and congenital malformations among surviving Atox1(-/-) progeny. Furthermore, Atox1-deficient cells accumulated high levels of intracellular copper, and metabolic studies indicated that this defect was because of impaired cellular copper efflux. Taken together, these data reveal a direct role for Atox1 in trafficking of intracellular copper to the secretory pathway of mammalian cells and demonstrate that this metallochaperone plays a critical role in perinatal copper homeostasis. PMID- 11391008 TI - A larger hippocampus is associated with longer-lasting spatial memory. AB - Volumetric studies in a range of animals (London taxi-drivers, polygynous male voles, nest-parasitic female cowbirds, and a number of food-storing birds) have shown that the size of the hippocampus, a brain region essential to learning and memory, is correlated with tasks involving an extra demand for spatial learning and memory. In this paper, we report the quantitative advantage that food storers gain from such an enlargement. Coal tits (Parus ater) a food-storing species, performed better than great tits (Parus major), a nonstoring species, on a task that assessed memory persistence but not on a task that assessed memory resolution or on one that tested memory capacity. These results show that the advantage to the food-storing species associated with an enlarged hippocampus is one of memory persistence. PMID- 11391007 TI - Selection for in vivo regulators of bacterial virulence. AB - We devised a noninvasive genetic selection strategy to identify positive regulators of bacterial virulence genes during actual infection of an intact animal host. This strategy combines random mutagenesis with a switch-like reporter of transcription that confers antibiotic resistance in the off state and sensitivity in the on state. Application of this technology to the human intestinal pathogen Vibrio cholerae identified several regulators of cholera toxin and a central virulence gene regulator that are operative during infection. These regulators function in chemotaxis, signaling pathways, transport across the cell envelope, biosynthesis, and adherence. We show that phenotypes that appear genetically independent in cell culture become interrelated in the host milieu. PMID- 11391009 TI - Autoimmunity to beta IV spectrin in paraneoplastic lower motor neuron syndrome. AB - Paraneoplastic neurological disorders may result from autoimmunity directed against antigens shared by the affected neurons and the associated cancer cells. We have recently reported the case of a woman with breast cancer and paraneoplastic lower motor neuron syndrome whose serum contained autoantibodies directed against axon initial segments and nodes of Ranvier of myelinated axons, including the axons of motoneurons. Here, we show that major targets of the autoantibodies of this patient are betaIVSigma1 spectrin and betaIV spectrin 140, two isoforms of the novel betaIV spectrin gene, as well as a neuronal surface epitope yet to be identified. Partial improvement of the neurological symptoms following cancer removal was associated with a drastic reduction in the titer of the autoantibodies against betaIV spectrin and nodal antigens in general, consistent with the autoimmune pathogenesis of the paraneoplastic lower motor neuron syndrome. The identification of betaIV spectrin isoforms and surface nodal antigens as novel autoimmune targets in lower motor neuron syndrome provide new insights into the pathogenesis of this severe neurological disease. PMID- 11391010 TI - PLS1, a gene encoding a tetraspanin-like protein, is required for penetration of rice leaf by the fungal pathogen Magnaporthe grisea. AB - We describe in this study punchless, a nonpathogenic mutant from the rice blast fungus M. grisea, obtained by plasmid-mediated insertional mutagenesis. As do most fungal plant pathogens, M. grisea differentiates an infection structure specialized for host penetration called the appressorium. We show that punchless differentiates appressoria that fail to breach either the leaf epidermis or artificial membranes such as cellophane. Cytological analysis of punchless appressoria shows that they have a cellular structure, turgor, and glycogen content similar to those of wild type before penetration, but that they are unable to differentiate penetration pegs. The inactivated gene, PLS1, encodes a putative integral membrane protein of 225 aa (Pls1p). A functional Pls1p-green fluorescent protein fusion protein was detected only in appressoria and was localized in plasma membranes and vacuoles. Pls1p is structurally related to the tetraspanin family. In animals, these proteins are components of membrane signaling complexes controlling cell differentiation, motility, and adhesion. We conclude that PLS1 controls an appressorial function essential for the penetration of the fungus into host leaves. PMID- 11391012 TI - Optimal classification of protein sequences and selection of representative sets from multiple alignments: application to homologous families and lessons for structural genomics. AB - Hierarchical classification is probably the most popular approach to group related proteins. However, there are a number of problems associated with its use for this purpose. One is that the resulting tree showing a nested sequence of groups may not be the most suitable representation of the data. Another is that visual inspection is the most common method to decide the most appropriate number of subsets from a tree. In fact, classification of proteins in general is bedevilled with the need for subjective thresholds to define group membership (e.g., 'significant' sequence identity for homologous families). Such arbitrariness is not only intellectually unsatisfying but also has important practical consequences. For instance, it hinders meaningful identification of protein targets for structural genomics. I describe an alternative approach to cluster related proteins without the need for an a priori threshold: one, through its use of dynamic programming, which is guaranteed to produce globally optimal solutions at all levels of partition granularity. Grouping proteins according to weights assigned to their aligned sequences makes it possible to delineate dynamically a 'core-periphery' structure within families. The 'core' of a protein family comprises the most typical sequences while the 'periphery' consists of the atypical ones. Further, a new sequence weighting scheme that combines the information in all the multiply aligned positions of an alignment in a novel way is put forward. Instead of averaging over all positions, this procedure takes into account directly the distribution of sequence variability along an alignment. The relationships between sequence weights and sequence identity are investigated for 168 families taken from HOMSTRAD, a database of protein structure alignments for homologous families. An exact solution is presented for the problem of how to select the most representative pair of sequences for a protein family. Extension of this approach by a greedy algorithm allows automatic identification of a minimal set of aligned sequences. The results of this analysis are available on the Web at http://mathbio.nimr.mrc.ac.uk/~amay. PMID- 11391013 TI - Use of a database of structural alignments and phylogenetic trees in investigating the relationship between sequence and structural variability among homologous proteins. AB - The database PALI (Phylogeny and ALIgnment of homologous protein structures) consists of families of protein domains of known three-dimensional (3D) structure. In a PALI family, every member has been structurally aligned with every other member (pairwise) and also simultaneous superposition (multiple) of all the members has been performed. The database also contains 3D structure-based and structure-dependent sequence similarity-based phylogenetic dendrograms for all the families. The PALI release used in the present analysis comprises 225 families derived largely from the HOMSTRAD and SCOP databases. The quality of the multiple rigid-body structural alignments in PALI was compared with that obtained from COMPARER, which encodes a procedure based on properties and relationships. The alignments from the two procedures agreed very well and variations are seen only in the low sequence similarity cases often in the loop regions. A validation of Direct Pairwise Alignment (DPA) between two proteins is provided by comparing it with Pairwise alignment extracted from Multiple Alignment of all the members in the family (PMA). In general, DPA and PMA are found to vary rarely. The ready availability of pairwise alignments allows the analysis of variations in structural distances as a function of sequence similarities and number of topologically equivalent Calpha atoms. The structural distance metric used in the analysis combines root mean square deviation (r.m.s.d.) and number of equivalences, and is shown to vary similarly to r.m.s.d. The correlation between sequence similarity and structural similarity is poor in pairs with low sequence similarities. A comparison of sequence and 3D structure-based phylogenies for all the families suggests that only a few families have a radical difference in the two kinds of dendrograms. The difference could occur when the sequence similarity among the homologues is low or when the structures are subjected to evolutionary pressure for the retention of function. The PALI database is expected to be useful in furthering our understanding of the relationship between sequences and structures of homologous proteins and their evolution. PMID- 11391014 TI - An approach to improving multiple alignments of protein sequences using predicted secondary structure. AB - The object of this work was to improve multiple sequence alignments using public domain software and methods as far as possible. A method is described where the secondary structure of proteins is predicted and this information, coupled with a simplified description of the amino acids, is used to produce multiple sequence alignments. This method improved the accuracy of the resulting alignments by between 5 and 14% when compared with full sequence profile alignments (as scored against structural alignments). These improved alignments were used to predict the secondary structure of the sequences they contain. The resultant predictions were more accurate than those produced from less optimal alignments. An improvement of 6% for a three-state (helix, sheet and coil) prediction was observed when using the best alignment from the method presented here and the alignment obtained using sequence only. The method makes use of public domain software and all the associated files required to repeat the work are available from the primary author. PMID- 11391015 TI - Modeling multi-component protein-DNA complexes: the role of bending and dimerization in the complex of p53 dimers with DNA. AB - We used molecular modeling to study the optimal conformation of the complex between two p53 DNA-binding domain monomers and a 12 base-pair target DNA sequence. The complex was constructed using experimental data on the monomer binding conformation and a new approach to deform the target DNA sequence. Combined with an internal/helicoidal coordinate model of DNA, this approach enables us to bend the target sequence in a controlled way while respecting the contacts formed with each p53 monomer. The results show that the dimeric complex favors DNA bending towards the major groove at the dimer junction by a value close to experimental findings. In contrast to inferences from earlier models, the calculation of key contributions to the free energy of the complexes indicates a determinant role for DNA in the formation of the complex with the dimer of the p53 DNA-binding domains. PMID- 11391016 TI - Mutant barley (1-->3,1-->4)-beta-glucan endohydrolases with enhanced thermostability. AB - The similar three-dimensional structures of barley (1-->3)-beta-glucan endohydrolases and (1-->3,1-->4)-beta-glucan endohydrolases indicate that the enzymes are closely related in evolutionary terms. However, the (1-->3)-beta glucanases hydrolyze polysaccharides of the type found in fungal cell walls and are members of the pathogenesis-related PR2 group of proteins, while the (1-->3,1 ->4)-beta-glucanases function in plant cell wall metabolism. The (1-->3)-beta glucanases have evolved to be significantly more stable than the (1-->3,1-->4) beta-glucanases, probably as a consequence of the hostile environments imposed upon the plant by invading microorganisms. In attempts to define the molecular basis for the differences in stability, eight amino acid substitutions were introduced into a barley (1-->3,1-->4)-beta-glucanase using site-directed mutagenesis of a cDNA that encodes the enzyme. The amino acid substitutions chosen were based on structural comparisons of the barley (1-->3)- and (1-->3,1- >4)-beta-glucanases and of other higher plant (1-->3)-beta-glucanases. Three of the resulting mutant enzymes showed increased thermostability compared with the wild-type (1-->3,1-->4)-beta-glucanase. The largest increase in stability was observed when the histidine at position 300 was changed to a proline (mutant H300P), a mutation that was likely to decrease the entropy of the unfolded state of the enzyme. Furthermore, the three amino acid substitutions which increased the thermostability of barley (1-->3,1-->4)-beta-glucanase isoenzyme EII were all located in the COOH-terminal loop of the enzyme. Thus, this loop represents a particularly unstable region of the enzyme and could be involved in the initiation of unfolding of the (1-->3,1-->4)-beta-glucanase at elevated temperatures. PMID- 11391017 TI - A single point mutation (Glu85Arg) increases the stability of the thioredoxin from Escherichia coli. AB - Glu85 in the Escherichia coli thioredoxin, which is localized in the loop between beta4 and beta5, was substituted with the Arg present in the corresponding position in Bacillus acidocaldarius thioredoxin. This suggested that it could play an important role in the structure and thermostability of this protein owing to its involvement in numerous interactions. The effects of the mutation on the biophysical properties were analysed by circular dichroism, spectrofluorimetry and limited proteolysis, supported by molecular dynamics data. As modelling predicted, an increase in stability for E85R due to additional H-bonds between the beta5 and alpha4 regions was observed. PMID- 11391018 TI - Application of a very high-throughput digital imaging screen to evolve the enzyme galactose oxidase. AB - Directed evolution has become an important enabling technology for the development of new enzymes in the chemical and pharmaceutical industries. Some of the most interesting substrates for these enzymes, such as polymers, have poor solubility or form highly viscous solutions and are therefore refractory to traditional high-throughput screens used in directed evolution. We combined digital imaging spectroscopy and a new solid-phase screening method to screen enzyme variants on problematic substrates highly efficiently and show here that the specific activity of the enzyme galactose oxidase can be improved using this technology. One of the variants we isolated, containing the mutation C383S, showed a 16-fold increase in activity, due in part to a 3-fold improvement in K(m). The present methodology should be applicable to the evolution of numerous other enzymes, including polysaccharide-modifying enzymes that could be used for the large-scale synthesis of modified polymers with novel chemical properties. PMID- 11391019 TI - Enantioselectivity of recombinant Rhizomucor miehei lipase in the ring opening of oxazolin-5(4H)-ones. AB - Enantioselectivity of enzyme catalysis is often rationalized via active site models. These models are constructed on the basis of comparing the enantiomeric excess of product observed in a series of reactions which are conducted with a range of homologous substrates, typically carrying various side chain substitutions. Surprisingly the practical application of these simple but informative 'pocket size' models has been rarely tested in genetic engineering experiments. In this paper we report the construction, purification and enantioselectivity of two recombinant Rhizomucor miehei lipases which were designed to check the validity of such a model in reactions of ring opening of oxazolin-5(4H)-ones. PMID- 11391020 TI - Truncated aspartate aminotransferase from alkalophilic Bacillus circulans with deletion of N-terminal 32 amino acids is a non-functional monomer in a partially structured state. AB - Aspartate aminotransferase (AspAT) from alkalophilic Bacillus circulans contains an additional N-terminal sequence of 32 amino acid residues that are absent in all other AspATs from different sources. Modeling suggested that this sequence forms two alpha-helical segments which establish a continuous network of interactions on the surface of the molecule. In the present study, we studied the role of the N-terminal sequence in folding and stability of AspAT by applying the scanning calorimetry, and CD and fluorescence spectroscopies to the native and truncated enzymes. Truncated AspAT (Delta2alpha mutant) devoid of N-terminal residues cannot provide sufficient potential of quaternary intersubunit and subunit-cofactor interactions, which results in a monomeric non-functional conformation. However, the residual tertiary interactions in the Delta2alpha mutant are sufficient to: i) provide stability of a residual structure over a wide pH range; ii) confer moderate cooperativity of the denaturant-induced transition while only low cooperativity of the thermal transition, and iii) maintain the hydrophobic core of a part of the structure which prevents aromatic fluorophores from quenching by water. Furthermore, the present study provides evidence that AspAT from the alkalophilic bacterium follows unfolding pathway comprising a stable non-functional intermediate, in contrast to a two-state mechanism of the thermophilic AspAT from Sulfolobus solfataricus. PMID- 11391021 TI - A single H:CDR3 residue in the anti-digoxin antibody 26-10 modulates specificity for C16-substituted digoxin analogs. AB - We constructed Fab libraries of bacteriophage-displayed H:CDR3 mutants in the high-affinity anti-digoxin antibody 26-10 to determine structural constraints on affinity and specificity for digoxin. Libraries of mutant Fabs randomized at five or 10 contiguous positions were panned against digoxin and three C16-substituted analogs, gitoxin (16-OH), 16-formylgitoxin and 16-acetylgitoxin. The sequence data from 83 different mutant Fabs showed highly restricted consensus patterns at positions H:100, 100a and 100b for binding to digoxin; these residues contact digoxin in the 26-10:digoxin co-crystal structure. Several mutant Fabs obtained following panning on digoxin-BSA showed increased affinity for digoxin compared with 26-10 and retained the wild-type (wt) Trp at position 100. Those Fabs selected following panning on C16-substituted analogs showed enhanced binding to the analogs. Replacement of H:Trp100 by Arg resulted in mutants that bound better to the analogs than to digoxin. This specificity change was unexpected, as C16 lies on the opposite side of digoxin from H:CDR3. Substitution of wt Trp by Arg appears to alter specificity by allowing the hapten to shift toward H:CDR3, thereby providing room for C16 substituents in the region of H:CDR1. PMID- 11391022 TI - A strategy for optimizing the monodispersity of fusion proteins: application to purification of recombinant HPV E6 oncoprotein. AB - Recombinant production of HPV oncoprotein E6 is notoriously difficult. The unfused sequence is produced in inclusion bodies. By contrast, fusions of E6 to the C-terminus of carrier proteins such as maltose-binding protein or glutathione S-transferase are produced soluble. However, it has not yet been possible to purify E6 protein from such fusion constructs. Here, we show that this was due to the biophysical heterogeneity of the fusion preparations. We find that soluble MBP-E6 preparations contain two subpopulations. A major fraction is aggregated and contains exclusively misfolded E6 moieties ('soluble inclusion bodies'). A minor fraction is monodisperse and contains the properly folded E6 moieties. Using monodispersity as a screening criterion, we optimized the expression conditions, the purification process and the sequence of E6, finally obtaining stable monodisperse MBP-E6 preparations. In contrast to aggregated MBP-E6, these preparations yielded fully soluble E6 after proteolytic removal of MBP. Once purified, these E6 proteins are stable, folded and biologically active. The first biophysical measurements on pure E6 were performed. This work shows that solubility is not a sufficient criterion to check that the passenger protein in a fusion construct is properly folded and active. By contrast, monodispersity appears as a better quality criterion. The monodispersity-based strategy presented here constitutes a general method to prepare fusion proteins with optimized folding and biological activity. PMID- 11391026 TI - MHC and the viral hepatitides. PMID- 11391027 TI - Current management and novel therapeutic strategies for refractory ascites and hepatorenal syndrome. AB - The circulatory disturbances seen in advanced cirrhosis lead to the development of ascites, which can become refractory to diet and medical therapy. These abnormalities may progress and cause a functional renal failure known as the hepatorenal syndrome. Management of refractory ascites and hepatorenal syndrome is a therapeutic challenge, and if appropriate, liver transplantation remains the best treatment. New therapeutic options have recently appeared, including the transjugular intrahepatic portosystemic shunt and selective splanchnic vasoconstrictor agents, which may improve renal function and act as a bridge to transplantation. PMID- 11391028 TI - Are patients suffering from stable angina receiving optimal medical treatment? AB - There is good evidence for the use of antiplatelet, beta-blocker and lipid lowering drugs in the treatment of ischaemic heart disease, but few data on how these medications are used in treating stable angina pectoris. We examined prescription profiles for a sample of patients aged > or =65 years with stable angina, to compare the profiles to local guidelines and to explore the determinants of these profiles, in a cross-sectional study. We identified 11 141 individuals from the Quebec provincial out-patient pharmaceutical database for the period 1 June 1996 to 31 May 1997, and examined the percentage of these patients with and without associated co-morbidities receiving antiplatelet, beta blocker and lipid-lowering medications. We used hierarchical modelling to examine the role of patient and physician characteristics in explaining the variation in the use of these medications. Calcium-channel blockers were the class of anti ischaemic drugs most prescribed (63%). Beta-blockers were prescribed in 52.1% of patients. Antiplatelet and lipid-lowering drugs were prescribed to 56.8% and 32.6%, respectively. Increasing age and female gender made patients less likely to be prescribed these treatments. General practitioners were less likely than cardiologists to prescribe beta-blockers and lipid-lowering drugs (OR 0.79, CI 95% 0.68-0.91 and OR 0.77, CI 95% 0.66-0.91, respectively). There is a general under-use of antiplatelet, beta-blocker and lipid-lowering medications in the treatment of stable angina pectoris patients, possibly leading to adverse patient outcomes. PMID- 11391029 TI - Adverse reactions to disease-modifying anti-rheumatic drugs in clinical practice. AB - We analysed computerized records of disease-modifying anti-rheumatic drug (DMARD) monotherapy to determine how long rheumatoid arthritis (RA) patients continued on five commonly prescribed DMARDs, and the incidence and time-course of adverse drug reactions (ADRs) they experienced. We studied the records for 3923 courses of DMARDs given to a cohort of 2170 patients monitored for a total of 9378 treatment-years. Methotrexate (MTX) was the DMARD most likely to be continued long-term; <45% of patients had discontinued the drug after 96 months. For the other DMARDs, the time until 50% discontinued due to ADRs or inefficacy was 43.3 months for sulphasalazine (SAS), 33.9 months for D-penicillamine (DPN) and 26 months for myocrisin. Most monitored ADRs requiring drug discontinuation were seen early in therapy, with a median time to onset of <6 months; the important exceptions to this were haematological ADRs to MTX, where the median delay to neutropenia was 16.9 months, and that to thrombocytopenia was 9.4 months. Monitored ADRs (identified by blood or urine tests) were seen least frequently with SAS (one ADR in every 35 patient-years of monitoring) but this apparent advantage was offset by a high incidence of gastrointestinal ADRs and inefficacy. Overall, one toxicity reaction requiring drug discontinuation was identified for every 15.9 patient-years of monitoring. PMID- 11391030 TI - Activation of the endothelin system in insulin resistance. AB - Endothelin-1, released from the vascular endothelium after cleavage from big endothelin-1, is a potent paracrine vasoconstrictor peptide. Small studies suggest that circulating levels of endothelin-1 are elevated in subjects with cardiovascular risk factors. Big endothelin-1 levels may better reflect endothelin-1 generation. We examined relationships between plasma endothelin-1, plasma big endothelin-1, and predisposition to hypertension or other cardiovascular risk factors associated with insulin resistance in a large group of healthy young men. We recruited 96 healthy men aged 24-33 years from a cohort of 864 young men and women in whom predisposition to hypertension had been defined on the basis of their own blood pressure and the blood pressures of their parents. They attended after an overnight fast for measurement of blood pressure, anthropometry, and plasma lipids, insulin, glucose, endothelin-1 and big endothelin-1. Plasma endothelin-1 and big endothelin-1 levels did not correlate with blood pressure (r=0.09, -0.002 respectively) and were not influenced by parental blood pressure. Higher plasma endothelin-1 levels were associated with higher body mass index (r=0.29, p<0.005), and higher plasma insulin (r=0.21, p<0.05). Higher plasma big endothelin-1 levels were associated with insulin resistance, as assessed by the Homeostasis Model of Assessment resistance index (r=0.30, p<0.005). Endothelin-1 levels are not related to blood pressure, but are higher in healthy young men with insulin resistance and obesity. PMID- 11391031 TI - Dyslipidaemia in patients with malignant-phase hypertension. AB - Low-density lipoprotein (LDL) consists of a heterogeneous group of particles of differing size, density and electrophoretic mobility, smaller particles being more atherogenic. A high proportion of small LDL particles is an independent risk factor for cardiovascular disease. We hypothesized that patients with malignant phase hypertension (MHT), the most severe form of hypertension, would demonstrate a more atherogenic LDL subfraction profile than either non-malignant hypertension (NMHT) or normotensive controls. We compared 16 patients with MHT to 41 patients with untreated NMHT and 45 normotensive controls. LDL subfraction profile was measured by disc polyacrylamide gel electrophoresis using a validated scoring system to calculate the mean size (locus) and heterogeneity (spread) of LDL subfraction mobilities. A higher LDL locus indicates a greater proportion of small LDL subfractions. LDL cholesterol levels were similar in all three groups (p=0.23). High-density lipoprotein cholesterol (HDL-C) levels were significantly lower (p<0.001) and serum triglyceride concentrations significantly higher (p=0.02) in the MHT group, compared to normotensive controls. LDL locus was greater in the NMHT group than in the normotensive controls and intermediate in the MHT group (p=0.008). There was no significant difference in LDL spread (p=0.26). Serum triglyceride concentrations were not significantly higher after adjusting for confounding variables. MHT is associated with an abnormal lipid profile, characterized by low HDL-cholesterol concentration. This dyslipidaemia may be partly responsible for the vascular complications and the poor prognosis of these patients. PMID- 11391033 TI - Surgical approaches to the management of heart failure. PMID- 11391032 TI - The potential for pharmacological treatment of unpleasant psychological symptoms to increase personal fulfillment in old age. AB - As some people get older, they experience a decline in their subjective sense of fulfillment. Life may become less rewarding, happiness diminished in intensity. This is usually regarded as an inevitable consequence of the ageing process: regrettable, but a circumstance to which stoical endurance is the only constructive response. This situation is potentially avoidable, for some individuals at least; not at some indefinite point in the future, but now. By using existing and available drugs in a novel fashion to treat the unpleasant psychological symptoms associated with ageing, a substantial improvement in the quality of life may be obtained. PMID- 11391035 TI - Mitral valve reconstruction in the patient with heart failure. AB - Secondary MR is a complication of end-stage cardiomyopathy and is associated with a poor prognosis and is due to progressive mitral annular dilation and alteration in LV geometry. A vicious cycle of continuing volume overload, ventricular dilation, progression of annular dilation, increased LV wall tension and worsening MR and CHF occur. The mainstay of medical therapy is diuretics and afterload reduction, and is associated with poor long-term survival in these patients with CHF and MR. However, surgical intervention in the form of undersized, 'overcorrecting' mitral valve repair has shown great promise and is an area of ongoing investigation. PMID- 11391034 TI - Myocardial revascularization as a therapeutic strategy in the patient with advanced ventricular dysfunction. AB - The number of patients with severe ventricular dysfunction from coronary artery disease is constantly increasing. Although the medical management of these patients with angiotensin-converting enzyme inhibitors and beta-blockers has favorable impact on the morbidity and mortality the overall prognosis is still poor. Historically many of these patients have been referred for transplantation. In the past few years there has been an increasing amount of information about the utility of surgical revascularization in patients with low ejection fraction. Careful patient selection and optimal perioperative management is of critical importance for good outcome. Coronary artery bypass grafting (CABG) can be performed relatively safely despite the advanced level of left ventricular dysfunction. Quality of life is improved by CABG with elimination of angina and enhanced functional capacity. Improvement in the ejection fraction and increased survival after the operation has been objectively demonstrated. However patients with advanced right ventricular dysfunction, pulmonary hypertension, redo bypass and ungraftable coronaries should be considered for heart transplantation. In this review we describe our experience and focus on pertinent issues in patient selection, perioperative management and long term outcome after coronary artery bypass grafting. PMID- 11391036 TI - The endoventricular circular patch plasty ("Dor procedure") in ischemic akinetic dilated ventricles. AB - From 1984 to 2000, 950 Left Ventricular ischemic asynergy (dyskinetic or akinetic) were operated using the endoventricular circular patch plasty technique. This allows to exclude all asynergic areas of the left ventricular wall and reshape the remaining wall. Both morphology and hemodynamic of left ventricle, are improved. Hospital mortality was below 7%. Life expectancy at 10 years reaches 80% if pre-operative L.V.E.F. is above 30%, and end systolic volume index (E.S.V.I.) below 90[emsp4 ]ml, and 60% in L.V.E.F. is below 30% and E.S.V.I. above 90[emsp4 ]ml. L.V.R. by endoventricular plasty has to be considered in the treatment of ischemic congestive heart failure. PMID- 11391037 TI - The Batista procedure: fact, fiction and its role in the management of heart failure. AB - End-stage heart failure is associated with high rates of mortality. Obviously, heart transplantation is the ultimate surgical intervention for its treatment. However, this surgical option is severely limited by immunosuppressive drug morbidity and inadequate donor organ availability. Partial left ventriculectomy, the so called Batista procedure, has been proposed for the treatment of dilated cardiomyopathy and other end-stage heart failure. Although initial reports lacked significant information on the safety and efficacy of this procedure, overall clinical impression from the reports is that the operation may serve as a relatively inexpensive bridge to transplantation especially in the patients with idiopathic dilated cardiomyopathy. In order to select an exact procedure to resect appropriate amount of scar tissue, dobutamine echocardiographic study, intraoperative volume reduction test using cardiopulmonary bypass, positron emission tomography, or magnetic resonance imaging scans can be used. To avoid the late deterioration related to the development of significant mitral valve regurgitation, definitive mitral valve repair or replacement at the time of the partial left ventriculectomy may be advised. Further study is required to determine the procedure's exact role in the treatment of congestive heart failure. This would have to be a multicenter, randomized, and long-term follow-up study. PMID- 11391038 TI - Dynamic cardiomyoplasty as a therapeutic alternative: current status. AB - Dynamic cardiomyoplasty was proposed as an alternative surgical treatment for severe cardiomyopathies and has been performed worldwide in more than 1,000 patients. Patients indicated for this procedure are specifically those with dilated or ischemic cardiomyopathies. The ventricular function improvement observed after dynamic cardiomyoplasty derived from the direct action of synchronized skeletal muscle flap contraction and from a girdling effect that helps to reverse chamber remodeling and to decrease ventricular wall stress. Although long-term benefits of this procedure may be limited by skeletal muscle flap ischemic compromise, technological advances incorporated in the new myostimulators will possibly decrease this complication incidence. Clinical improvement has been reported as a consistent finding in cardiomyoplasty follow up and the overall 5-year survival after this procedure ranges from 39 % to 54 %. On the other hand, the mortality after cardiomyoplasty has been significantly higher for patients in persistent New York Heart Association functional class IV, showing that this procedure needs to be indicated earlier than the heart transplantation. In this regard, only the results of an ongoing randomized trial will potentially define cardiomyoplasty influence on the survival of patients with severe heart failure. In the meantime, however, there are clearly several functional class III patients whose quality of life and exercise capacity have worsened despite the use of maximum medical therapy, justifying dynamic cardiomyoplasty indication. PMID- 11391039 TI - Heart transplantation--surgical results. AB - Heart transplantation has become a widely used therapeutic option for the treatment of end-stage heart failure. Since the first human orthotopic heart transplant in the late 1960s, the surgical technique has undergone several revisions. These revisions have addressed certain anatomic and geometric distortions that occurred with the original biatrial technique of Lower and Shumway. Early revisions have included the use of a bicaval technique for implanting the right atrium. Subsequently, the additional use of a direct pulmonary venous anastomosis has lead to the surgical concept of the total orthotopic heart transplant. These revisions of the original bi-atrial technique have led to a decrease in atrial size and distortion, conduction abnormalities and tricuspid and mitral valve regurgitation. This has also resulted in less atrial thromboembolic events, less need for permanent postoperative pacemaker placement, and in an overall increase in right and left heart performance in the early postoperative period. Overall, this has contributed to better clinical results with patients returning sooner to their normal exercise capacity. Ninety percent of heart transplant patients lead a relatively normal lifestyle having no limitations in their activity and 40 % return to work. We believe that the technique of total orthotopic heart transplantation has improved surgical results and clinical outcomes. PMID- 11391040 TI - Heart transplantation in the young and elderly. AB - INTRODUCTION: Heart transplantation has become an acceptable treatment in pediatric patients with end-stage heart disease and complex congenital heart disease. The liberalization of recipient eligibility criteria, mainly age, along with the expansion of the donor pool has resulted in the acceptable transplantation of older recipients. METHODS: Between July 1994 and June 1998, 39 pediatric patients aged 16 days to 17.6 years (median 6.68 years) and 123 elderly patients aged 60 to 74.8 years (median 64.1 years) were transplanted at our institution. In the pediatric group, 19 had idiopathic dilated cardiomyopathy (DCM) (46 %), 14 had congenital heart disease (34 %), 4 had other etiologies of cardiomyopathy (10 %), 2 had transplant coronary artery disease (TCAD) (5 %), and 1 each had acute rejection and graft failure. In the elderly group, 71 had ischemic cardiomyopathy (58 %), 38 had DCM (31 %), 9 had other forms of cardiomyopathy (7 %), and 5 had TCAD (4 %). RESULTS: Thirty-day, 1-year, and 4 year survival was 97.4 %, 87.2 %, and 70.9 % for the pediatric group and 92.7 %, 81.3 %, and 79.3 % for the elderly group. One and 4-year freedom from TCAD was 100.0 % and 85.3 % for the pediatric group and 91.9 % and 83.3 % for the elderly group. CONCLUSIONS: [emsp3 ]Orthotopic heart transplantation is effective for the treatment of irreparable congenital and end-stage heart disease. It provides excellent long-term results in both the very young and elderly. PMID- 11391043 TI - The protein kinase C family and lipid mediators for transmembrane signaling and cell regulation. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. PMID- 11391041 TI - The challenge of rejection and cardiac allograft vasculopathy. AB - Since the first human heart transplantation was performed in 1967, the field of heart transplantation has advanced to the point where survival and acceptable quality of life are commonplace. Despite remarkable progress in the clinical management of rejection, rejection continues to limit survival and quality of life in the heart transplant population. This review will discuss the biologic processes involved in hyperacute rejection, acute rejection, and humoral (vascular) rejection. The development of endomyocardial biopsy techniques represented a significant advancement in the diagnosis of cardiac rejection, and endomyocardial biopsy remains the 'gold standard' in the diagnosis of cellular rejection. To date, no noninvasive parameters will diagnose rejection with adequate sensitivity and specificity. Biopsy frequency and immunosuppressive therapies may be tailored to the risk of rejection. Immunosuppression for cardiac transplantation can be divided into three major phases: 1) perioperative immunosuppression; 2) maintenance immunosuppression, and; 3) treatment of rejection. The strategy for treating transplant rejection should be influenced by several variables: 1) Histologic grade of rejection; 2) Evidence of hemodynamic compromise by ejection fraction or right heart catheterization; 3) Severity of previous rejection episodes and types of immunosuppressives used; and 4) Risk factors for rejection, including time after transplantation. Future rejection therapy will involve more sophisticated attempts to alter host responses toward the donor organ in a more specific and selective way. Despite considerable advances in the care of the heart transplant recipient, long-term survival is limited by cardiac allograft vasculopathy. The final section of this chapter will review the pathology, immunopathology, nonimmunologic risk factors, diagnosis, prevention and treatment of allograft vasculopathy. PMID- 11391044 TI - Nonalcoholic fatty liver disease: implications for alcoholic liver disease pathogenesis. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The presentation was Nonalcoholic fatty liver disease: Implications for alcoholic liver disease pathogenesis, by Anna Mae Diehl. PMID- 11391045 TI - The role of acetaldehyde in the actions of alcohol (update 2000). AB - BACKGROUND: Recent advances in the field of acetaldehyde (AcH) research have raised the need for a comprehensive review on the role of AcH in the actions of alcohol. This update is an attempt to summarize the available AcH research. METHODS: The descriptive part of this article covers not only recent research but also the development of the field. Special emphasis is placed on mechanistic analyses, new hypotheses, and conclusions. RESULTS: Elevated AcH during alcohol intoxication causes alcohol sensitivity, which involves vasodilation associated with increased skin temperature, subjective feelings of hotness and facial flushing, increased heart and respiration rate, lowered blood pressure, sensation of dry mouth or throat associated with bronchoconstriction and allergy reactions, nausea and headache, and also reinforcing reactions like euphoria. These effects seem to involve catecholamine, opiate peptide, prostaglandin, histamine, and/or kinin mechanisms. The contribution of AcH to the pathological consequences of chronic alcohol intake is well established for different forms of cancer in the digestive tract and the upper airways. AcH seems to play a role in the etiology of liver cirrhosis. AcH may have a role in other pathological developments, which include brain damage, cardiomyopathy, pancreatitis, and fetal alcohol syndrome. AcH creates both unpleasant aversive reactions that protect against excessive alcohol drinking and euphoric sensations that may reinforce alcohol drinking. The protective effect of AcH may be used in future treatments that involve gene therapy with or without liver transplantation. CONCLUSIONS: AcH plays a role in most of the actions of alcohol. The individual variability in these AcH-mediated actions will depend on the genetic polymorphism, not only for the alcohol and AcH metabolizing enzymes but also for the target sites for AcH actions. The subtle balance between aversive and reinforcing, protecting and promoting factors will determine the overall behavioral and pathological developments. PMID- 11391046 TI - Ethanol and lipid metabolic signaling. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Shivendra D. Shukla and Grace Y. Sun. The presentations were (1) Metabolic turnover of ethanol into cellular lipids and platelet activating factor, by Shivendra D. Shukla; (2) Ethanol action on the phospholipase A2 signaling pathways in astrocytes, by Grace Y. Sun; (3) Mechanisms of ethanol-induced perturbation of lipoprotein cholesterol transport, by W. Gibson Wood; (4) Transfer of an abnormal ethanol-induced phospholipid, phosphatidylethanol, between lipoproteins, by Markku J. Savolainen; (5) Phospholipase-d-mediated formation of phosphatidylethanol, by Christer Alling; and (6) Changes in phosphoinositide signaling after chronic ethanol treatment, by Jan B. Hoek. PMID- 11391047 TI - Sex difference in alcohol-related organ injury. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Nobuhiro Sato and Kai O. Lindros. The presentations were (1) Sex differences in ethanol pharmacokinetics, by E. Baraona; (2) Estrogen regulates the sensitivity to endotoxin in hepatic Kupffer cells, by K. Ikejima; (3) Sex difference in alcohol-related organ injury, by E. Mezey; (4) Aggravated ethanol-induced liver injury in female rats: Protection by the antiestrogen toremifene, by Harri A. Jarvelainen; and (5) Alcohol metabolism in Asian subjects: Sex differences and flushing response, by V. A. Ramchandani. PMID- 11391048 TI - cAmp signaling cascade: a promising role in ethanol tolerance and dependence. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Subhash C. Pandey and Toshikazu Saito. The presentations were (1) Action of ethanol on cAMP signaling pathways, by M. Yoshimura; (2) Alterations in the G protein adenylyl cyclase system and their mRNA levels in alcoholics, by H. Sohma; (3) The role of the CREB gene transcription factor in ethanol dependence and preference, by Subhash C. Pandey; and (4) The efficacy of adenylyl cyclase signal transduction to the nucleus in primary alcoholics, by M. E. Gotz. PMID- 11391049 TI - Alcoholic liver disease and apoptosis. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Carol A. Casey and Amin Nanji. The presentations were (1) Mechanisms of apoptosis in alcoholic liver disease, by Amin A. Nanji; (2) Impaired receptor-mediated endocytosis: Its role in alcoholic apoptosis, by Carol A. Casey; (3) Toxicity of ethanol in HepG2 cells that express CYP2E1, by Arthur I. Cederbaum; (4) Mitochondrial regulation of ethanol-induced hepatocyte apoptosis, by M. Adachi; and (5) Apoptosis in alcoholic hepatitis, by T. Takahashi. PMID- 11391050 TI - Recent advances in the pathology of alcoholic myopathy. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Victor R. Preedy and Junko Adachi. The presentations were (1) Alcoholic myopathy: Past, present and future, by Timothy J. Peters and Victor R. Preedy; (2) Protein adducts in the type I and II fiber predominant muscles of the ethanol-fed rat, by Simon Worrall, Seppo Parkkila, and Onni Niemela; (3) Hydroperoxides and changes in alcoholic myopathy, by Junko Adachi, Migiwa Asamo, and Yasuhino Ueno; and (4) A close association between testicular atrophy, muscle atrophy, and the increase in protein catabolism after chronic ethanol administration, by Kunihiko Takeda, Masayoshi Yamauchi, Kazuhiko Sakamoto, Masaru Takagi, Hisato Nakajima, and Gotaro Toda. PMID- 11391051 TI - Transgenic and gene "knockout" models in alcohol research. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Paula L. Hoffman and Takeshi Yagi. The presentations were (1) cAMP signaling in ethanol sensitivity and tolerance, by Boris Tabakoff; (2) Synaptic signaling pathways of Fyn-tyrosine kinase, by Takeshi Yagi; (3) Ethanol drinking and sensitization in dopaminergic and serotonergic receptor knockouts, by Tamara J. Phillips; (4) ICAM-1 is involved in early alcohol-induced liver injury in the mouse given enteral alcohol, by Hiroshi Kono; and (5) Strategies for targeted and regulated knockouts, by Robert O. Messing and Doo-Sup Choi. PMID- 11391052 TI - Alcohol biomarkers: clinical significance and biochemical basis. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Raj Lakshman and Mikihiro Tsutsumi. The presentations were (1) Sialic acid index of apolipoprotein J: A new marker for chronic alcohol consumption, by P. Ghosh and M. R. Lakshman; (2) Microheterogeneity of serum glycoproteins in alcoholics, by M. Tsutsumi and S. Takase; (3) Probing protein-ethanol adducts with combinatorial peptide libraries displayed by filamentous phage, by H. Anni, O. Nikolaeva, and Y. Israel Y; (4) Carbohydrate-deficient transferrin as a marker for heavy alcohol use: What have we learned; Where do we go from here, by R. F. Anton; (5) Sensitivity and specificity of carbohydrate-deficient transferrin in drinking experiments and different patient groups, by O. M. Lesch; (6), Transferrin variants interfere with the measurement of carbohydrate-deficient transferrin, by A. Helender, G. Eriksson, and J-O. Jeppson; and (7) Chronic ethanol on protein trafficking in liver, by P. Marmillot, M. N. Rao, and M. R. Lakshman. PMID- 11391053 TI - Metabolic and ethnic determinants of alcohol drinking habits and vulnerability to alcohol-related disorder. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Shoji Harada and Dharam P. Agarwal. The presentations were (1) Mutations in the exons, exon-intron junctions, and promoter regions of human CYP2E1 gene and alcoholism, by Fumio Nomura; (2) Genetic variability in alcohol metabolism and drinking habits in Japanese, by Shoji Harada; (3) Genetic studies of alcohol dependence using alcoholics with inactive ALDH2, by Susumu Higuchi; and (4) Alcohol consumption, apolipoprotein polymorphisms, and cardiovascular disorders, by Dharam P. Agarwal. PMID- 11391054 TI - Alcohol actions on GABA(A) receptors: from protein structure to mouse behavior. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were R. Adron Harris and Susumu Ueno. The presentations were (1) Protein kinase Cepsilon-regulated sensitivity of gamma aminobutyric acid type A (GABAA) receptors to allosteric agonists, by Robert O. Messing, A. M. Sanchez-Perez, C. W. Hodge, T. McMahon, D. Wang, K. K. Mehmert, S. P. Kelley, A. Haywood, and M. F. Olive; (2) Genetic and functional analysis of a GABAA receptor gamma2 subunit variant: A candidate for quantitative trait loci involved in alcohol sensitivity and withdrawal, by Kari J. Buck and Heather M. Hood; (3) Tryptophan-scanning mutagenesis in GABAA receptor subunits: Channel gating and alcohol actions, by Susumu Ueno; and (4) Can a single binding site account for actions of alcohols on GABAA and glycine receptors? by R. Adron Harris, Yuri Blednov, Geoffrey Findlay, and Maria Paola Mascia. PMID- 11391055 TI - Ethanol and gene expression in brain. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Izuru Matusmoto and Peter A. Wilce. The presentations were (1) GABA receptor subunit expression in the human alcoholic brain, by Tracey Buckley and Peter Dodd; (2) NMDAR gene expression during ethanol addiction, by Jorg Puzke, Rainer Spanagel, Walther Zieglgansberger, and Gerald Wolf; (3) Differentially expressed gene in the nucleus accumbens from ethanol administered rat, by Shuangying Leng; (4) Expression of a novel gene in the alcoholic brain, by Peter A. Wilce; and (5) Investigations of haplotypes of the dopamine D2-receptor gene in alcoholics, by Hans Rommelspacher, Ulrich Finckh, and Lutz G. Schmidt. PMID- 11391056 TI - Core heritable personality characteristics and relapse in alcoholics. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were David S. Janowsky and Jan Fawcett. The presentations were (1) The tridimensional personality questionnaire: Predictor of relapse in detoxified alcoholics, by Kurt Meszaros; (2) Novelty seeking predicts clinical trial attrition in alcoholics, by Jan Fawcett; (3) Personality and alcohol/substance use disorder patient relapse and attendance at self-help group meetings, by David S. Janowsky; and (4) A three-pathway psychobiological model for craving for alcohol, by Roel Verheul. PMID- 11391057 TI - WHP/ISBRA study on state and trait markers in alcoholism. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was Boris Tabakoff. The presentations were (1) Overview of the WHO/ISBRA study on state and trait markers in alcoholism, by Boris Tabakoff; (2) Biochemical markers of acute and chronic drinking: Results of the WHO/ISBRA study, by Anders Helander; (3) The impact of country of recruitment and body mass index on biological marker dose-response curves in the WHO/ISBRA Study, by Kate M. Conigrave; (4) Relationship of body water to carbohydrate deficient transferrin measures, by Larry Martinez; and (5) Platelet adenylyl cyclase activity as a trait marker of alcohol dependence, by Paula L. Hoffman. PMID- 11391058 TI - Neuroimaging in alcoholism: ethanol and brain damage. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The co-chairs were Karl Mann and Ingrid Agartz. The presentations were (1) Neuropathological changes in alcohol-related brain damage, by Clive Harper; (2) Regional brain volumes including the hippocampus and monoamine metabolites in alcohol dependence, by Ingrid Agartz, Susan Shoaf, Robert R, Rawlings, Reza Momenan, and Daniel W Hommer; (3) Diffusion tensor abnormalities in imaging of white matter alcoholism, by Adolf Pfefferbaum and Edith V. Sullivan; (4) Use of functional MRI to evaluate brain activity during alcohol cue exposure in alcoholics: Relationship to craving, by Raymond F. Anton, David J. Drobes, and Mark S. George; and (5) mu-Opiate receptor availability in alcoholism: First results from a positron emission tomography study, by Karl Mann, Roland Bares, Hans-Juergen Machulla, Goetz Mundle, Matthias Reimold, and Andreas Heinz. PMID- 11391059 TI - Fetal alcohol effects: mechanisms and treatment. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was Edward P. Riley. The presentations were (1) Does alcohol withdrawal contribute to fetal alcohol effects? by Jennifer D. Thomas and Edward P. Riley; (2) Brain damage and neuroplasticity in an animal model of binge alcohol exposure during the "third trimester equivalent," by Charles R. Goodlett, Anna Y. Klintsova, and William T. Greenough; (3) Ganglioside GM1 reduces fetal alcohol effects, by Basalingappa L. Hungund; and (4) Fetal alcohol exposure alters the wiring of serotonin system at mid-gestation, by F. Zhou, Y. Sari, Charles Goodlett, T. Powrozek, and Ting-Kai Li. PMID- 11391060 TI - Alcohol reinforcement and voluntary ethanol consumption. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The organizer/chair was Ting-Kai Li and the co-chair was Rainer Spanagel. The presentations were (1) Genetic differences in alcohol drinking and reinforcement: The sP and sNP Rats, by Giancarlo Colombo; (2) Ventral tegmental area-Neuroanatomical substrate for alcohol reinforcement, by William J. McBride; (3) Metabolic mapping of alcohol reinforcement, by Linda J. Porrino; (4) Role of opioid receptors in the ethanol-induced place preference in rats exposed to conditioned fear stress, by Tsutomu Suzuki; and (5) Repeated deprivations enhance the reinforcing properties of ethanol in alcohol preferring (P) rats, by Zachary A. Rodd-Henricks. PMID- 11391061 TI - Pharmacological relapse prevention in alcohol dependence: from animal models to clinical trials. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Jobst August-Ludwig Boening and Otto Michel Lesch. The presentations were (1) Pharmacological validation of a new animal model of alcoholism, by Rainer Spanagel; (2) Persisting loss of control as main criterion for alcohol addiction in rats and mice, by Jochen Wolffgramm; (3) Role of NMDA receptor subunits associated with protein kinase C in the prevention of alcohol dependence, by Minoru Narita; (4) Long-term follow up of continued naltrexone treatment, by David Sinclair; (5) Pharmacological treatment trials with dopaminergic and serotonergic substances: Myths or facts? by Gerhard A. Wiesbeck; and (6) Methodology and behavioral therapy of the U.S. acamprosate study, by Barbara J. Mason. PMID- 11391062 TI - Role of polyamines and NMDA receptors in ethanol dependence and withdrawal. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was John M. Littleton. The presentations were (1) Examination of ethanol spermine and acamprosate actions on native and recombinant NMDA receptors, by David Lovinger; (2) Ethanol inhibition of NMDA neurotoxicity on the polyamine site in cerebellar granule cells, by Sture Liljequist; (3) Alterations in expression of NMDA receptor subunits during ethanol exposure and withdrawal, by Raj Ticku; (4) Alterations in polyamine synthesis and release as a potential mechanism for ethanol dependence and withdrawal, by Izuru Matsumoto; (5) The role of polyamines in neurotoxicity induced by alcohol withdrawal in vitro, by John Littleton; and (6) Agmatine reduces some of the effects of "third trimester" alcohol exposure using a rodent model, by Susan Barron. PMID- 11391063 TI - Alcohol and cancer. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Helmut K. Seitz and Shohei Matsuzaki. The presentations were (1) Alcohol dehydrogenase-2 and aldehyde dehydrogenase-2 genotype and cancer risk for upper aerodigestive tract in Japanese alcoholics, by Akira Yokoyama; (2) The role of acetaldehyde in alcohol-associated carcinogenesis, by Nils Homann; (3) High salivary acetaldehyde levels after a moderate dose of alcohol in ALDH2-deficient subjects, by Satu Vakevainen; (4) Alcohol and vitamin A interactions, by Xian Dong Wang; and (5) Alcohol and colorectal cancer, by Helmut K. Seitz. PMID- 11391064 TI - Neurobiological processes in alcohol addiction. AB - This article represents the proceedings of a symposium at the ISBRA Meeting in Yokohama, Japan. The chairs were A. D. Le and K. Kiianmaa. The presentations were (1) Alcohol reward and aversion, by C. L. Cunningham; (2) The role of sensitization of neuronal mechanisms in ethanol self-administration, by J. A. Engel, M. Ericson, and B. Soderpalm; (3) Alcohol self-administration in dependent animals: Neurobiological mechanisms, by G. F. Koob, A. J. Roberts, and F. Weiss; (4) Stress and relapse to alcohol, by A. D. Le; (5) Alcohol-preferring AA and alcohol-avoiding ANA rats differ in locomotor activation induced by repeated morphine injections, by P. Hyytia, S. Janhunen, J. Mikkola, P. Backstrom, and K. Kiianmaa; and (6) Initial sensitivity and acute functional tolerance to the hypnotic effects of ethanol in mice genetically selected for mild and severe ethanol withdrawal convulsions, by I. Ponomarev and J. C. Crabbe. PMID- 11391065 TI - Mechanisms of alcohol-nicotine interactions: alcoholics versus smokers. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Toshio Narahashi and Bo Soderpalm. The presentations were (1) Nicotinic mechanisms and ethanol reinforcement: Behavioral and neurochemical studies, by Bo Soderpalm, M. Ericson, P. Olausson, and J. A. Engel; (2) Chronic nicotine and ethanol: Differential regulation in gene expression of nicotinic acetylcholine receptor subunits, by X. Zhang and A. Nordberg; (3) Nicotine-ethanol interactions at neuronal nicotinic acetylcholine receptors, by Toshio Narahashi, William Marszalec, and Gary L. Aistrup; (4) Relapse prevention in alcoholics by cigarette smoking? Treatment outcome in an observational study with acamprosate, by L.G. Schmidt, U. Kalouti, M. Smolka, and M. Soyka; and (5) Effect of nicotine on voluntary ethanol intake and development of alcohol dependence in male rats, by L. Hedlund and G. Wahlstrom. PMID- 11391066 TI - Functional relevance of human adh polymorphism. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were C. J. Peter Eriksson and Tatsushige Fukunaga. The presentations were (1) 4-Methylpyrazole as a tool in the investigation of the role of ADH in the actions of alcohol in humans, by Taisto Sarkola and C. J. Peter Eriksson; (2) ADH2 polymorphism and flushing in Asian populations, by Wei J. Chen, C. C. Chen, J. M. Ju, and Andrew T. A. Cheng; (3) Role of ADH3 genotypes in the acute effects of alcohol in a Finnish population, by Hidetaka Yamamoto, Kathrin Kohlenberg-Muller, and C. J. Peter Eriksson; (4) Clinical characteristics and disease course of alcoholics with different ADH2 genotypes, by Mitsuru Kimura, Masanobu Murayama, Sachio Matsushita, Haruo Kashima, and Susumu Higuchi; (5) ADH2 polymorphism, alcohol drinking, and birth defects, by Lucinda Carr, D. Viljoen, L. Brooke, T. Stewart, T. Foroud, J. Su, and Ting-Kai Li; and (6) ADH genotypes and alcohol use in Europeans, by John B. Whitfield. PMID- 11391067 TI - Perioperative morbidity and mortality in chronic alcoholic patients. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Claudia Spies and Hanne Tonnesen. The presentations were (1) Relevance of alcohol misuse in surgical patients, by Hanne Tonnesen; (2) Diagnosis of alcohol abuse and alcohol dependence, by Sven Andreasson; (3) Diagnosis of acute alcohol misuse, by Anders Helander; (4) Preoperative intervention for excessive alcohol consumption, by Kate Conigrave; and (5) Prevention and treatment of perioperative complications in chronic alcoholics, by Claudia Spies. PMID- 11391069 TI - Neuroreceptors and ion channels as targets of alcohol. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Toshio Narahashi and Kinya Kuriyama. The presentations were (1) Modulation of neuroreceptors and ion channels by alcohol, by T. Narahashi; (2) Inhibition by ethanol of NMDA and AMPA receptor-channels, by P. Illes, K. Wirkner, W. Fischer, K. Muhlberg, P. Scheibler, and C. Allgaier; (3) Effects of ethanol on metabotropic glutamate receptors, by K. Minami; (4) Acute alcohol actions on the 5-HT3 ligand-gated ion channel, by D. Lovinger; (5) Inhibition of NMDA receptors by MK801 attenuates ethanol-induced taurine release from the hippocampus, by F. Lallemand, R.J. Ward, and P. DeWitte; and (6) Effect of ethanol on voltage-operated Ca2+ channels in hepatic stellate cells, by T. Itatsu, Y. Takei, H. Oide, M. Hirose, X. E. Wang, S. Watanabe, M. Tateyama, R. Ochi, and N. Sato. PMID- 11391068 TI - How is the liver primed or sensitized for alcoholic liver disease? AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Hidekazu Tsukamoto and Yoshiyuki Takei. The presentations were (1) Tribute to Professor Rajendar K. Chawla, by Craig J. McClain; (2) Dysregulated TNF signaling in alcoholic liver disease, by Craig J. McClain, S. Joshi-Barve, D. Hill, J Schmidt, I. Deaciuc, and S. Barve; (3) The role of mitochondria in ethanol-mediated sensitization of the liver, by Anna Colell, Carmen Garcia-Ruiz, Neil Kaplowitz, and Jose C. Fernandez-Checa; (4) A peroxisome proliferator (bezafibrate) can prevent superoxide anion release into hepatic sinusoid after acute ethanol administration, by Hirokazu Yokoyama, Yukishige Okamura, Yuji Nakamura, and Hiromasa Ishii; (5) S-adenosylmethionine affects tumor necrosis factor-alpha gene expression in macrophages, by Rajendar K. Chawla, S. Barve, S. Joshi-Barve, W. Watson, W. Nelson, and C. McClain; (6) Iron, retinoic acid and hepatic macrophage TNFalpha gene expression in ALD, by Hidekazu Tsukamoto, Min Lin, Mitsuru Ohata, and Kenta Motomura; and (7) Role of Kupffer cells and gut-derived endotoxin in alcoholic liver injury, by N. Enomoto, K. Ikejima, T. Kitamura, H. Oide, Y. Takei, M. Hirose, B. U. Bradford, C. A. Rivera, H. Kono, S. Peter, S. Yamashina, A. Konno, M. Ishikawa, H. Shimizu, N. Sato, and R. Thurman. PMID- 11391070 TI - Recent research on alcohol tolerance and dependence. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Hiroshi Suwaki and Harold Kalant. The presentations were (1) Influence of ADH genotypes on acute alcohol withdrawal syndrome in Japanese, by Susumu Higuchi; (2) Use of genetic analyses to refine phenotypes related to alcohol tolerance and dependence, by John C. Crabbe; (3) Neurochemical basis for alcohol dependence, by Seitaro Ohkuma and Masashi Katsura; (4) Adenylyl cyclase and development of tolerance to addictive drugs, by Masami Yoshimura; (5) Tolerance in rat lines selectively bred for alcohol preference, by Robert C. Stewart and Ting-Kai Li; and (6) Ethanol reinforcement, dependence, and vulnerability to relapse: Interactions between neuroadaptive and conditioning factors, by Friedbert Weiss. PMID- 11391071 TI - Alcohol withdrawal kindling: mechanisms and implications for treatment. AB - This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was Larry P. Gonzalez. The presentations were (1) EEG indices of sensitization in a murine model of repeated ethanol withdrawals, by Lynn M. Veatch; (2) Long-term changes in central nervous system function after repeated alcohol withdrawals: Recommendations for the treatment of acute withdrawal, by Larry P. Gonzalez; (3) Differential regulation of GABAA and NMDA receptors by repeated ethanol treatment in cultured mammalian neurons, by Maharaj K. Ticku; and (4) Involvement of GABAA and NMDA receptors in alcohol withdrawal kindling: Implications for treatment, by Howard C. Becker. PMID- 11391072 TI - Fetal alcohol syndrome: an international perspective. AB - This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Kenneth R. Warren and Faye J. Calhoun. The presentations were (1) Epidemiological research on fetal alcohol syndrome (FAS) in the United States, by Philip A. May; (2) An overview of fetal alcohol syndrome in the Western Cape Province of South Africa, by Denis L. Viljoen and Ting-Kai Li; (3) Diagnostic perspectives of fetal alcohol and tobacco syndromes, by Harumi Tanaka; (4) FAS among pupils of special boarding schools and orphanages in Moscow, Russia, by Galina S. Marinicheva and Luther K. Robinson; and (5) Research on FAS and FAE in Germany: Update and perspectives, by Goetz Mundle. PMID- 11391073 TI - Alcohol and retinoids. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Hirokazu Yokoyama and David Crabb. The presentations were (1) Roles of vitamin A, retinoic acid, and retinoid receptors in the expression of liver ALDH2, by J. Pinaire, R. Hasanadka, M. Fang, and David W. Crabb; (2) Alcohol, vitamin A, and beta-carotene: Adverse interactions, by M. A. Leo and Charles S. Lieber; (3) Retinoic acid, hepatic stellate cells, and Kupffer cells, by Hidekazu Tsukamoto, K. Motomura, T. Miyahara, and M. Ohata; (4) Retinoid storage and metabolism in liver, by William Bosron, S. Sanghani, and N. Kedishvili; (5) Characterization of oxidation pathway from retinol to retinoic acid in esophageal mucosa, by Haruko Shiraishi, Hirokazu Yokoyama, Michiko Miyagi, and Hiromasa Ishii; and (6) Ethanol in an inhibitor of the cytosolic oxidation of retinol in the liver and the large intestine of rats as well as in the human colon mucosa, by Ina Bergheim, Ina Menzl, Alexandr Parlesak, and Christiane Bode. PMID- 11391074 TI - The chemistry and biological effects of malondialdehyde-acetaldehyde adducts. AB - This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Geoffrey M. Thiele and Simon Worrall. The presentations were (1) The chemistry of malondialdehyde-acetaldehyde (MAA) adducts, by Dean J. Tuma; (2) The formation and clearance of MAA adducts in ethanol-fed rats, by Simon Worrall; (3) Immune responses to MAA adducts may play a role in the development of alcoholic liver disease, by Lynell W. Klassen; (4) Unique biological responses to MAA-modified proteins that may play a role in the development and/or progression of alcoholic liver disease, by Geoffrey M. Thiele; (5) MAA-adducted bovine serum albumin activates protein kinase C and stimulates interleukin-8 release in bovine bronchial epithelial cells, by Todd A. Wyatt; and (6) An enzyme immune assay for serum antiacetaldehyde adduct antibody using low density lipoprotein-adduct and its significance in alcoholic liver injury and ALDH2 heterozygotes, by Naruhiko Nagata. PMID- 11391075 TI - The ubiquitin-proteasome 26s pathway in liver cell protein turnover: effect of ethanol and drugs. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Samuel W. French and R. J. Mayer. The presentations were (1) The ubiquitin-proteasome 26s pathway in liver cell protein turnover: Effect of alcohol and drugs, by Samuel W. French and F. Bardag-Gorce; (2) The role of CYP2E1 phosphorylation and degradation pathway in the induction of the enzyme, by Magnus Ingelman-Sundberg; (3) Role of proteasome in the proteolysis of oxidized proteins in experimental chronic alcoholism, by Helen Rouach; (4) Alcohol, proteolysis and liver cancer, by R. J. Mayer; (5) Effect of ethanol feeding on the ATP-ubiquitin-proteasome pathway in the liver cell, by F. Bardag-Gorce; (6) Novel mechanisms and targets for intracellular transport of CYP2E1, by E. Neve; and (7) Gankyrin, an oncoprotein commonly over expressed in hepatoma, by H. Higashitsuji. PMID- 11391076 TI - Role of estrogen in alcohol promotion of breast cancer and prolactinomas. AB - This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was Dipak K. Sarkar. The presentations were (1) Dual role of estrogen as hormone and carcinogen in mammary carcinogenesis, by Joachim G. Liehr; (2) Alcohol and breast cancer: Studies using animals, by Keith W. Singletary; and (3) Evaluation of the role of estrogen in mediation of ethanol effect on prolactinoma: Studies using animals, by Dipak K. Sarkar. PMID- 11391077 TI - Ethanol and oxidative stress. AB - This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was Albert Y. Sun. The presentations were (1) Ethanol-inducible cytochrome P-4502E1 in alcoholic liver disease, by Magnus Ingelman-Sundberg and Etienne Neve; (2) Regulation of NF-kappaB by ethanol, by H. Matsumoto, Y. Nishitani, Y. Minowa, and Y. Fukui; (3) Chronic ethanol consumption increases concentration of oxidized proteins in rat liver, by Shannon M. Bailey, Vinood B. Patel, and Carol C. Cunningham; (4) Antiphospholipids antibodies and oxidized modified low-density lipoprotein in chronic alcoholic patients, by Tomas Zima, Lenka Fialova, Ludmila Mikulikova, Ptr Popov, Ivan Malbohan, Marta Janebova, and Karel Nespor; and (5) Amelioration of ethanol-induced damage by polyphenols, by Albert Y. Sun and Grace Y. Sun. PMID- 11391078 TI - Alcohol and the pancreas. AB - This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The presentations were (1) Phenotypic alteration of myofibroblast during ethanol-induced pancreatic injury: its relation to bFGF, by Masahiko Nakamura, Kanji Tsuchimoto, and Hiromasa Ishii; (2) Activation of pancreatic stellate cells in pancreatic fibrosis, by Paul S. Haber, Gregory W. Keogh, Minoti V. Apte, Corey S. Moran, Nancy L. Stewart, Darrell H.G. Crawford, Romano C. Pirola, Geoffrey W. McCaughan, Grant A. Ramm, and Jeremy S. Wilson; (3) Pancreatic blood flow and pancreatic enzyme secretion on acute ethanol infusion in anesthetized RAT, by H. Nishino, M. Kohno, R. Aizawa, and N. Tajima; (4) Genotype difference of alcohol-metabolizing enzymes in relation to chronic alcoholic pancreatitis between the alcoholic in the National Institute on Alcoholism and patients in other general hospitals in Japan, by K. Maruyama, H. Takahashi, S. Matsushita, K. Okuyama, A. Yokoyama, Y. Nakamura, K. Shirakura, and H. Ishii; and (5) Alcohol consumption and incidence of type 2 diabetes, by Katherine M. Conigrave, B. Frank Hu, Carlos A. Camargo Jr, Meir J. Stampfer, Walter C. Willett, and Eric B. Rimm. PMID- 11391080 TI - Models of alcoholic liver disease in rodents: a critical evaluation. AB - This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were J. Christian Bode and Hiroshi Fukui. The presentations were (1) Essentials and the course of the pathological spectrum of alcoholic liver disease in humans, by P. de la M. Hall; (2) Lieber-DeCarli liquid diet for alcohol-induced liver injury in rats, by C. S. Lieber and L. M. DeCarli; (3) Tsukamoto-French model of alcoholic liver injury, by S. W. French; (4) Animal models to study endotoxin-ethanol interactions, by K. O. Lindros and H. Jarvelainen; and (5) Jejunoileal bypass operation in rats-A model for alcohol induced liver injury? by Christiane Bode, Alexandr Parlesak, and J. Christian Bode. PMID- 11391079 TI - Mechanisms of alcoholic liver disease: cytokines. AB - This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chair was Manuela G. Neuman. The presentations were (1) New aspects of hepatic fibrosis, by D. A. Brenner; (2) Cellular immune response in hepatitis C models, by B. Rehermann; (3) The role of interleukin-10 in acute alcoholic hepatitis, by J. Taieb, S. Chollet-Martin, M. Cohard, J. J. Garaud, and T. Poynard; (4) Cytokine-mediated apoptosis in vitro, by M. G. Neuman; (5) Signaling for apoptosis and repair in vitro, by G. G. Katz, R. G. Cameron, N. H. Shear, and M. G. Neuman; (6) Interferons activate the P42/44 mitogen-activated protein kinase and Janus Kinase signal transducers and activation of transcription (JAK-STAT) signaling pathways in hepatocytes: Differential regulation by acute ethanol via a protein kinase C-dependent mechanism, by B. Gao; (7) Genetic polymorphisms of interleukin-1 in association with the development of Japanese alcoholic liver disease, by M. Takamatsu, M. Yamauchi, M. Ohata, S. Saito, S. Maeyama, T. Uchikoshi, and G. Toda; and (8) Increased levels of macrophage migration inhibitory factor in sera from patients with alcoholic liver diseases, by T. Kumagi, S. M. F. Akbar, M. Abe, K. Michitaka, N. Horiike, and M. Onji. PMID- 11391081 TI - Ethanol and protein metabolism. AB - This article represents the proceedings of a workshop at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Carol C. Cunningham and Victor R. Preedy. The presentations were (1) Ribosomal content, ribosomal localization and the levels of ribosomal protein mRNA and rRNA in rat skeletal muscle exposed to ethanol, by Alistair G. Paice, John E. Hesketh, Timothy J. Peters, and Victor R. Preedy; (2) Altered hepatic mitochondrial ribosome structure after chronic ethanol administration, by Vinood B. Patel and Carol C. Cunningham; (3) Clinical aspects of hepatic protein metabolism and alcohol, by Elena Volpi; and (4) Effects of oral intake of alanine plus glutamine on ethanol metabolism and ethanol-related depression in motor activity, by Kazunori Mawatari, H. Masaki, M. Mori, and Kunio Torii. PMID- 11391089 TI - A focus group on cognition-enhancing medications in Alzheimer disease: disparities between professionals and consumers. AB - The emergence of cognition-enhancing drugs in the treatment of Alzheimer disease raises questions about quality of lives for those with dementia and for their caregivers, and about the perceptions of health care professionals. This pilot study analyzes a limited data from a series of three focus groups on the experience of treatment. These groups engaged both Alzheimer disease-affected persons, their caregivers, and a multidisciplinary professional core. We conclude that therapeutic goals need to be better addressed with patients and families, as well as better monitored, with the possibility of withdrawing therapy as appropriate. We also detected, as hypothesized, considerable disparity between the perspectives of professionals and consumers regarding the benefits of therapy. PMID- 11391090 TI - The disability assessment for dementia scale: a 12-month study of functional ability in mild to moderate severity Alzheimer disease. AB - The Disability Assessment for Dementia (DAD) scale was developed and validated as a measure of functional ability in dementia. DAD results have been reported in Alzheimer disease (AD) randomized, controlled treatment trials of up to 6 months, but results beyond 6 months have yet to be described. SAB INT 12 was a randomized, double-blind, placebo-controlled, parallel-group study in mild to moderate AD that included DAD assessments at baseline, month 6, and month 12. One hundred forty-four patients with AD in the placebo arm of SAB INT 12 were followed up for 12 months. DAD scores were obtained at baseline (mean DAD = 70.1, SD = 22.2), 6 months (mean DAD = 63.7, SD = 25.2), and 12 months (mean DAD = 59.3, SD = 28.9). The rate of decline was consistent across the domains of basic activities of daily living (ADLs) and instrumental ADLs, as well as the scoring of initiation, planning, and organization. The decline in DAD total scores in mild to moderate AD averages about one point per month, which equates to the loss of one item on the DAD scale every 2 months. PMID- 11391094 TI - Morphological analysis of nevoid melanoma: a study of 20 cases with a review of the literature. AB - Nevoid melanoma is a rare variant of melanoma characterized by deceptive morphologic features reminiscent of a benign melanocytic nevus. Twenty (13 nodular, 7 verrucous) nevoid melanomas were reviewed with the goal of identifying the predominant architectural patterns, cytologic features, and prognostic indicators. Although at scanning magnification, many lesions showed a strong resemblance to banal compound or dermal nevi, careful inspection in all cases demonstrated subtle pleomorphism and impaired maturation with depth, invariably accompanied by multiple dermal mitoses. Four tumors recurred and three metastasized, with subsequent death of the patients. Follow-up information for a period of at least 3 years was available in eight cases. In this group, mortality was 37.5%, the metastasis rate was 37.5%, and the local recurrence rate was 75%, with an average tumor thickness of 2.5 mm. We conclude that nevoid melanoma may be distinguished from a benign melanocytic nevus by a high index of suspicion, a careful analysis of architecture, and attention to cytologic features. Our data and a review of the literature do not support the notion that nevoid melanoma has a better prognosis than ordinary melanoma. PMID- 11391095 TI - Interphase cytogenetic analysis of 1q12 satellite III DNA in melanocytic lesions: increased aneuploidy with malignant histology. AB - To examine the relationship of chromosome 1 copy number to melanocytic tumorigenesis, interphase cytogenetic analysis of 1q12 satellite III DNA was performed on the spectrum of melanocytic lesions comprising Clark's tumor progression model. Results showed increased copy number in a "step off" pattern between melanoma in-situ and the intraepidermal component of invasive melanoma rather than a progression between each lesional group. These findings support Clark's concept of independent clonal expansion of a cell population giving rise to the vertical growth phase and further demonstrates increased chromosome 1 copy number as a late event in melanoma tumor progression. PMID- 11391096 TI - Cutaneous B-cell lymphoma with signet ring-cell morphology: a clinicopathologic and immunohistochemical study of three cases. AB - Three cases of primary cutaneous B-cell lymphoma with prominent signet ring-cell features are presented. The patients were three men between the ages of 37 years and 74 years (average, 55.5 years). Clinically, the three patients presented with multiple skin nodules. In one patient, the nodules had been present for approximately 5 weeks, although in the two other patients, the nodules were of unknown duration. The lesions were located in the upper extremities (forearm) and measured from 2 cm to 3 cm in diameter. No evidence of lymphadenopathy was observed in any of the patients. Surgical excision of the nodules was performed. Histologically, in two cases, the superficial and deep dermis was replaced by a diffuse cellular proliferation, and in one patient, the tumor cell population adopted a nodular pattern of growth involving adnexal structures and infiltrating the subcutaneous fat. In all cases, the tumors were composed of cells showing signet ring-cell features, with striking indentation of the nuclei toward the periphery of the cell. Immunohistochemical studies using antibodies for B-cell and T-cell markers (L-26 and UCHL) as well as antibodies for leukocyte common antigen, keratin, and kappa and lambda light chains were performed in all cases. The tumor cells showed a positive reaction for leukocyte common antigen, L-26, and lambda light chain restriction. Follow-up information was only available in one patient, who has remained alive and well 2 years after diagnosis without evidence of progression of the disease. The present cases highlight the importance of recognizing this unusual morphologic type of lymphoma so as to arrive at a correct diagnosis. PMID- 11391097 TI - Microphthalmia transcription factor: not a sensitive or specific marker for the diagnosis of desmoplastic melanoma and spindle cell (non-desmoplastic) melanoma. AB - Microphthalmia transcription factor (Mitf), a melanocytic nuclear protein critical for the embryonic development and postnatal viability of melanocytes, is a master lineage regulator and modulates extracellular signals. Recently, Mitf expression was shown to be both a sensitive and specific marker of epithelioid melanoma. Because loss of specific melanocytic markers in melanomas with spindle cell morphology is more common compared with those tumors with epithelioid morphology, we investigated the sensitivity of D5, an anti-Mitf antibody, for diagnosis in this diagnostically problematic subset of melanomas. Twenty of 21 (95%) spindle cell and desmoplastic melanomas examined were reactive for S-100 protein. Only 4 of 21 (19%) spindle cell and desmoplastic melanomas were reactive for HMB-45. Six of 21 tumors (29%) were reactive for D5, including one case that was non-reactive for S-100 and HMB-45. Melan-A reactivity was seen in 2 of 13 cases (15%) studied. Eight of 24 (33%) non-melanocytic spindle cell tumors were reactive for D5, including 4 of 6 dermatofibromas, 1 of 6 schwannomas, 1 of 2 leiomyomas, and 2 of 6 leiomyosarcomas. Although D5 was shown in a previous study to be a highly sensitive and specific marker for epithelioid melanomas, the results of this study show it is not a sensitive or specific marker of spindle cell and desmoplastic melanomas. Nevertheless, we believe that diffuse positive staining for D5 when taken in clinical, histologic and immunohistochemical context may be diagnostically useful in selected cases of melanoma. PMID- 11391098 TI - The clinical and histologic spectrum of cutaneous fibrous perineuriomas. AB - Cutaneous fibrous perineuriomas (CFPs) and the closely related sclerotic perineuriomas are recently reported tumors. We present nine additional cases of CFPs. All tumors were small nodules on the extremities; however, they were not limited to a distal acral location as previously reported. In addition to tabulation of the histologic features, we performed a battery of immunohistochemical stains, including S-100 protein, CD34, cytokeratin, epithelial membrane antigen (EMA), KP-1, and collagen type IV. Histologically, these tumors presented with sharp circumscription of their deep aspect, as previously reported, or were not circumscribed on any side. The cells ranged from plump and spindled with one or more nuclei to thin, elongated, spindled cells with slender nuclei. A variably fibrotic stromal matrix surrounded these components. Immunohistochemical staining showed EMA-positive staining of the cellular component, with collagen type IV-positive staining surrounding the cells. The tumor cells were negative for S-100 protein, factor XIIIa, CD34, cytokeratin, and KP-1. The tumors reported here add to the clinical and histopathologic spectrum of CFPs. PMID- 11391099 TI - Adverse reactions to injectable aesthetic microimplants. AB - New inert materials such as polymerized silicones, Bioplastique, Artecoll, and Dermalive are now being used as injectable aesthetic microimplants. These substances are better than the old ones because they tend not to migrate and do not usually produce much of a host immune response. Adverse reactions after injection of these materials are rare, although there are a few reported cases as a result of bad technique or anomalous granulomatous reactions. We report on four patients with unsightly results after cosmetic microimplants, including one of Artecoll, one of Dermalive (to the best of our knowledge, the latter is the first such case reported), and two of silicone. This report describes the histopathologic features of cutaneous reactions to these injectable aesthetic materials. PMID- 11391100 TI - Basal cell carcinoma of the buccal mucosa. AB - True basal cell carcinoma (BCC) involving the oral mucous membranes is extraordinarily rare. Most of those described as occurring in the oral cavity usually involve the gingiva and are not true BCCs but peripheral ameloblastomas. A true BCC, which arose on the buccal mucosa of a 69-year old man, is reported. It presented as a 1.3 cm ulcerated plaque without gingival connection. Histologically, the lesion exhibited classic features of BCC with palisading and retraction spaces, and focally communicated with the overlying squamous epithelium. Although it is possible that this lesion also arose from a heterotopic odontogenic rest, the anatomical location, focal squamous (metatypical) features, and positive staining for Ber-EP4 support an origin from the basal cell layer of stratified squamous mucosa. PMID- 11391101 TI - Subcutaneous panniculitis-like T-cell lymphoma: an elusive case presenting as lipomembranous panniculitis and a review of 72 cases in the literature. AB - We present a remarkable case of subcutaneous panniculitic T-cell lymphoma (SPTL) that eluded diagnosis for 14 years and illustrates the importance of continued follow-up with repeat biopsy when SPTL is suspected. This case is unusual in that multiple biopsies demonstrated either a nonspecific panniculitis or lipomembranous panniculitis with calcified lipomembranes. A clinicopathologic review of 72 cases of SPTL from the English language literature is also presented, and approaches to diagnosis and treatment are reviewed. PMID- 11391102 TI - Zosteriform and epidermotropic metastatic primary cutaneous squamous cell carcinoma. AB - The first case of primary cutaneous squamous cell carcinoma (SCC) to cause zosteriform and epidermotropic metastasis to skin is reported. The patient is a 72-year-old Japanese woman. A cutaneous SCC appeared on the lateral side of her right knee and was removed. After dissection of the right inguinal lymph nodes, which revealed metastases, and irradiation of the right inguinal region, the patient presented with slightly pruritic and painful erythematous papules on the right hip and small brownish papules and vesicles with crusts on the anterior side of the right thigh. The eruptions were in a zosteriform distribution along the right L1 to L3 dermatomes. Histologically neoplastic squamous cell nests were observed in the epidermis, below the epidermal-dermal junction, and within lymphatic vessels in the deeper reticular dermis. We postulate that neoplastic cells with the ability to fuse with adjacent squamous epithelium may have been carried beneath the basal lamina or to the epidermis via dermal lymphatic backflow, resulting in epidermotropic metastasis. PMID- 11391103 TI - Trauma-induced simulator of targetoid hemosiderotic hemangioma. AB - Reported here is a 15-year-old with lesions demonstrating histologic features of targetoid hemosiderotic hemangioma (THH) developing after trauma to inflammatory lesions. These lesions pose as simulators of THH. Targetoid hemosiderotic hemangioma is a benign vascular lesion first described by Santa Cruz and Aaronburg. It classically presents as a single, small, red/brown, targetoid lesion on the trunk or extremities of a young or middle-aged individual. Histologically, it is characterized by ectatic vascular lumina in the papillary dermis lined by a single layer of endothelial cells with an epithelioid or "hobnail" appearance. In the deeper dermis, vascular spaces become slit-like and angulated, appearing to dissect through collagen bundles. A commonly proposed etiology of THH is trauma to a preexisting hemangioma. This case is remarkable for its unusual clinical presentation, histologic simulation of THH, and for its support for the theory that trauma can induce the histologic changes seen in THH. PMID- 11391104 TI - Pseudoepitheliomatous hyperplasia secondary to cutaneous aspergillus. AB - Cutaneous aspergillosis commonly occurs in immunocompromised hosts and may also complicate burn wounds. Pseudoepitheliomatous hyperplasia (PH) is a histologic reaction secondary to a wide range of stimuli, including fungal infection. We describe a case of an 18-year-old man, status-post burns over 70% of his total body surface area, with cutaneous aspergillosis of the axilla and secondary PH. A single case of PH secondary to primary aspergillosis has been described in the larynx but, to our knowledge, has never been described cutaneously. Histologic examination of the lesion reveals an irregularly acanthotic epidermis with deep invaginations within the dermis. There is an intense inflammatory reaction within the superficial and deep dermis. Numerous fungal forms are identified within the dermis. Special stains demonstrate septate hyphae with dichotomous branching, which is morphologically consistent with Aspergillus. Therefore, we conclude that cutaneous aspergillosis should be included in the differential diagnosis of causes of PH, especially in a patient population at risk for this infection. PMID- 11391105 TI - Atrichia with papular lesions: electron microscopic observations of cystic lesions. AB - Atrichia with papular lesions is a rare inherited skin disorder characterized by congenital atrichia with numerous papules. We describe a 27-year-old woman with atrichia, who had numerous papules on her scalp, nape, and axillae. Histologically, many keratinous cysts were seen in the middermis of a skin specimen from the nape. Electron microscopy showed that the developing keratinocytes in the walls of some cysts were rich in glycogen granules and had epidermoid keratinization with formation of keratohyaline granules and that laminated bodies were formed before keratinization. Langerhans cells were often seen in the walls of the cysts. In addition, a broad glassy vitreous layer surrounded the cyst wall. From these findings, it was suggested that the cystic lesions might have originated from immature or incomplete hair follicles. In particular, the structure of the cyst wall corresponded well to infundibular and/or isthmal portions of the outer root sheath of the hair follicle. PMID- 11391106 TI - The 21st Colloquium of the International Society of Dermatopathology. PMID- 11391107 TI - Neoplasms of the skin: benign vs. malignant. Editor-in-Chief's response to Dr. Weyers. PMID- 11391108 TI - Neoplasms of the skin: benign vs. malignant. Another response to Dr. Weyers. PMID- 11391109 TI - Tutorial on melanocytic lesions. PMID- 11391110 TI - Pitfalls in the histopathologic diagnosis of malignant melanoma. PMID- 11391111 TI - Serious limitations of a method. PMID- 11391112 TI - The 21st Colloquium of the International Society of Dermatopathology: Symposium on Melanocytic Lesions. PMID- 11391113 TI - Winer's dilated pore: the infundibuloma. AB - This article summarizes Louis H. Winer's description of the dilated pore and some of the pertinent literature about the dilated pore. To better understand the discussion that follows, there is a brief review of the histology of the follicular infundibulum and isthmus, then a discussion of the architectural and cytologic differentiation of proliferations that reputedly differentiate toward the infundibulum. These other proliferations with their original photomicrographs are discussed rather extensively to support the argument that they do not differentiate either architecturally or cellularly toward the infundibulum. I then present evidence that contradicts those authors who claim that Winer's dilated pore is only a cyst and conclude that it is a neoplasm sui generis and the only neoplasm of the skin that differentiates architecturally and cytologically towards the infundibulum (the infundibuloma). PMID- 11391114 TI - Louis H. Winer, M.D: the man behind the eponyms. PMID- 11391115 TI - Cutaneous mucinoses: microscopic criteria for diagnosis. AB - The clinical aspects and the histologic features of cutaneous mucinoses have been reviewed and their classification updated. Cutaneous mucinoses are divided into distinctive (primary) cutaneous mucinoses in which the mucin deposit is the main histologic feature resulting in clinically distinctive lesions, and disorders associated with histologic mucin deposition as an additional finding (secondary mucinoses). The former are further divided into degenerative-inflammatory mucinoses, which may be either dermal or follicular, and into neoplastic hamartomatous mucinoses. Histopathologic diagnosis is particularly difficult for dermal mucinoses and requires clinicopathologic correlation. Three histologic clues, namely the pattern of mucin distribution (diffuse or focal), the level of mucin deposit in the dermis and some additional findings may help diagnosis. Follicular mucinoses have the easiest pattern to recognize histologically, but the distinction between Pinkus' follicular mucinosis and follicular mucinosis with mycosis fungoides is very difficult.Lastly, neoplastic-hamartomatous cutaneous mucinoses include mucinous nevus, a benign hamartoma, and myxoma, which is a benign tumor to be differentiated from reactive cutaneous focal mucinosis. PMID- 11391116 TI - Proliferation of sweat ducts in a melanocytic neoplasm. PMID- 11391117 TI - Standards. PMID- 11391118 TI - The different originality of Homer and Thucydides. PMID- 11391119 TI - Solitary fibrous tumor of the skin. PMID- 11391120 TI - Histological and clinical overlapping. PMID- 11391121 TI - Gilles de la Tourette Syndrome. PMID- 11391122 TI - Platelet glutamate receptor supersensitivity in major depressive disorder. AB - Dysregulation of glutamate has been described in depression, and supersensitivity of platelet glutamate receptors has been found in both psychotic major depression and schizophrenia. The aim of this study was to examine the platelet glutamate receptor sensitivity in patients with nonpsychotic, unipolar major depression to assess whether this is a marker of depression or of psychosis. Glutamate receptor sensitivity was assessed using the platelet intracellular calcium response to glutamate (0-100 micromol) measured by spectrofluorometry. The depression group showed a significantly greater platelet intracellular calcium response to glutamate stimulation than the control group, both in terms of absolute values (p = 0.007) and percentage of response from baseline (p = 0.030). These data suggest that platelet glutamate receptors may be supersensitive in depression and that the platelet may be a possible peripheral marker of glutamate function in depression. PMID- 11391123 TI - A randomized, double-blind, placebo-controlled, ascending-dose tolerability and safety study of remacemide as adjuvant therapy in Parkinson's disease with response fluctuations. AB - The objective of this study was to establish the maximum tolerated dose of the low affinity non-competitive N-methyl-D-aspartate receptor antagonist remacemide in patients who have Parkinson's disease with response fluctuations or dyskinesias, or both. A total of 33 patients were randomly assigned in a 3-to-1 ratio to receive remacemide or placebo. Remacemide was administered orally at 150 mg twice daily, increasing incrementally by 100 mg (50 mg twice daily) at 2-week intervals to a final daily regimen of 400 mg twice daily or until a maximum tolerated dose was identified. The maximum total treatment period was 12 weeks. Of the 23 patients randomly selected to receive remacemide, four completed the study at the maximum permitted dose, compared with four of the 10 patients given placebo. The median maximum tolerated dose of remacemide was 450 mg/d. There was no clinically relevant change in percentage of "on" time between baseline and maximum tolerated dose in either group. At the maximum tolerated dose of remacemide for each patient, the mean Unified Parkinson's Disease Rating Scale (UPDRS) motor examination score (part III) decreased from 33 (SD = 18) to 26 (SD = 13) compared with a decrease from 28 (SD = 12) to 27 (SD = 8) in the placebo group. There was a decrease in the mean UPDRS "complications of therapy" score (part IV) in the remacemide group from 8 (SD = 4) to 6 (SD = 4), and the placebo group remained unchanged at 6 (SD = 4). The most common adverse events associated with remacemide were nausea, vomiting, dizziness, headache, abnormal vision, and hypokinesia. Remacemide was well tolerated at a dose level of 400 mg/d. There was a trend suggesting that remacemide was effective in improving symptoms at patients' individual maximum tolerated doses. These improvements occurred without exacerbating levodopa-induced dyskinesias. PMID- 11391124 TI - Cardiovascular effects of 0.5 milligrams per kilogram oral d-amphetamine and possible attenuation by haloperidol. AB - In a series of earlier studies, an oral dose of 0.5 mg/kg d-amphetamine was administered to 81 patients with schizophrenia and eight normal control subjects. Seven more subjects with schizophrenia received placebo. Blood pressure and pulse rate were monitored before and 3 hours after drug administration. Blood pressure increased in both amphetamine groups, whereas placebo had no effect. However, pulse rate did not change in the schizophrenic group and only increased after 3 hours in normal control subjects as blood pressure began to decrease. Significant negative correlations between systolic blood pressure and pulse rate occurred at 2 and 3 hours, suggesting that the early cardiovascular response to amphetamine is an increase in blood pressure that recruits reflex control of heart rate. Eighteen of these subjects had hypertensive responses. Six subjects received 5 mg haloperidol intramuscularly, and 12 others had their blood pressure monitored until normalization. Haloperidol led to a more rapid decline of some but not all indices of blood pressure, suggesting that amphetamine-induced hypertension may have a dopaminergic component. PMID- 11391125 TI - Double-blind, crossover, placebo-controlled trial of bromocriptine in patients with sleep bruxism. AB - This study was designed to assess the effects of bromocriptine, a dopamine D2 receptor agonist, on sleep bruxism. Seven otherwise healthy patients with severe and frequent sleep bruxism participated in this randomized, double-blind, placebo controlled study. The study used a crossover design that included 2 weeks of active treatment or placebo with a washout period of 1 week. To further evaluate whether bromocriptine influences striatal D2 receptor binding, we used iodine-123 iodobenzamide single photon emission computed tomography (SPECT) under both placebo and bromocriptine regimens. Bromocriptine did not reduce the frequency of episodes of bruxism during sleep (mean +/- SEM, 9.0 +/- 1.0 and 9.6 +/- 1.5 bruxism episodes per hour for placebo and bromocriptine, respectively) or the amplitude of masseter muscle contractions (root mean square values, 48.2 +/- 15.5 microV and 46.9 +/- 12.7 microV for placebo and bromocriptine, respectively). SPECT also failed to reveal that either treatment had any influence on striatal D2 binding (values for total binding in counts/pixel, 1.80 [1.72-1.93] and 1.79 [1.56-1.87] for placebo and bromocriptine, respectively). This study shows that a nightly dose of bromocriptine does not exacerbate or reduce sleep bruxism motor activity. PMID- 11391127 TI - QTc interval prolongation associated with citalopram overdose: a case report and literature review. AB - Citalopram is a member of the selective serotonin reuptake inhibitor class of antidepressants. In 1998, citalopram was approved by the US Food and Drug Administration for the treatment of major depression. Like the other selective serotonin reuptake inhibitors, citalopram enjoys a relatively benign side effect profile compared with the tricyclic antidepressants and the monoamine oxidase inhibitors. However, citalopram has been associated with electrocardiographic changes and seizures at doses greater than 600 mg per day. Fatalities have occurred with citalopram-only overdoses. We report the case of a healthy 21-year old woman who developed QTc interval prolongation after ingestion of approximately 400 mg citalopram. We discuss the cardiac effects of citalopram, review previous cases of citalopram overdose, and discuss treatment recommendations. PMID- 11391126 TI - The effects of different repeated doses of entacapone on the pharmacokinetics of L-Dopa and on the clinical response to L-Dopa in Parkinson's disease. AB - We performed a double-blind, placebo-controlled, randomized, crossover, multiple dose study on entacapone in 25 patients with Parkinson's disease with levodopa (L Dopa) treatment-related fluctuations. A run-in period was followed by four 2-week treatment periods during which the patients took 4 to 6 daily doses of L-Dopa concomitantly with 100, 200, or 400 mg of entacapone or with placebo. The effects were assessed at the end of each period; the inhibition of soluble catechol-O methyltransferase (S-COMT) activity in red blood cells and the plasma concentrations of entacapone, L-Dopa, and 3-O-methyldopa (3-OMD) were measured and clinical effects assessed on an 18-hour home diary. Twenty-one patients completed the study. Entacapone decreased the COMT activity from predose level: 100 mg by 25%, 200 mg by 33%, and 400 mg by 32% (p < 0.001 vs. placebo for each dose). Correspondingly, the 3-OMD concentrations decreased by 39%, 54%, and 66% with 100-, 200-, and 400-mg doses, respectively. The elimination half-life of L Dopa was prolonged by 23% (p < 0.05), 26% (p < 0.001), and 48% (p < 0.001), and the area under the curve of L-Dopa increased by 17% (p < 0.05), 27% (p < 0.001), and 37% (p < 0.001) with the increasing doses. Despite a significant decrease in the daily dose of L-Dopa, entacapone decreased the proportion of daily "off" time: 100 mg by 11%, 200 mg by 18%, and 400 mg by 20% compared with placebo. However, this decrease was not statistically significant for any of the doses in this small patient population. The dyskinetic "on" time did not increase with different doses of entacapone. All doses were well tolerated, and no severe adverse events were reported. The study showed that repeated dosing of entacapone inhibits the COMT activity in a dose-dependent manner and thereby reduces the loss of L-Dopa to 3-OMD. Therefore, the area under the curve of L-Dopa is increased and the patient's clinical condition improved. PMID- 11391128 TI - Continuous transdermal dopaminergic stimulation in advanced Parkinson's disease. AB - The objective of the study was to determine the safety and efficacy of increasing doses of Rotigotine CDS in patients with advanced Parkinson's disease. The development of motor complications in Parkinson's disease has been linked to intermittent stimulation of dopamine receptors. Continuous, noninvasive, dopaminergic stimulation has not been available to date. Rotigotine CDS is a lipid-soluble D2 dopamine agonist in a transdermal delivery system that could fill this void. This inpatient study consisted of a 2-week dose escalation phase followed by a 2-week dose maintenance phase at the highest dose (80 cm2). Each individual's L-Dopa dose was back-titrated as feasible. The primary outcome measure was L-Dopa dose, and secondary outcome measures included early morning "off"-L-Dopa Unified Parkinson's Disease Rating Scale motor scores by a blinded evaluator and motor fluctuation data obtained from patient diaries ("on" without dyskinesia, "on" with dyskinesia, and "off"). Seven of 10 subjects provided data that could be evaluated. There were two administrative dropouts, and one individual was eliminated from the study because of recrudescence of hallucinations. The median daily L-Dopa dose decreased from 1,400 to 400 mg (p = 0.018, Wilcoxon test). Unified Parkinson's Disease Rating Scale motor scores were unchanged. Although diary variables improved in most individuals, only the reduction in "off" time attained statistical significance. Adverse effects were mild and consisted mainly of dopaminergic side effects and local skin reactions. The data suggest that Rotigotine CDS is an effective treatment for advanced Parkinson's disease and permits patients to substantially lower L-Dopa doses without loss of antiparkinsonian efficacy. Full-scale controlled clinical trials are warranted. In addition to potential therapeutic benefits, this drug can be used to test the hypothesis that continuous dopaminergic stimulation from the initiation of Parkinson's disease therapy will limit the development of motor complications. PMID- 11391129 TI - Pathologic gambling in patients with Parkinson's disease. AB - Patients with Parkinson's disease frequently have depression, anxiety, and obsessive-compulsive disorder. We observed two patients who had episodes of pathologic gambling. At the same time, their Parkinson's disease deteriorated and they initiated self-medication with dopaminergic drugs. In both patients, signs were present of an addiction to dopaminergic medication. Pathologic gambling ceased in these patients after a few months. The significance of an insufficient dopaminergic reward system in patients with stereotypical addictive-like behavior (e.g., pathologic gambling) is discussed in this report. The most likely explanation for this newly recognized behavioral disorder in patients with Parkinson's disease is enhanced novelty seeking as a consequence of overstimulation of mesolimbic dopamine receptors resulting from addiction to dopaminergic drugs. PMID- 11391130 TI - The use of intravenous immunoglobulin as maintenance therapy in myasthenia gravis. AB - The standard therapy for myasthenia gravis (MG) includes steroids and immunosuppressants, which have delayed onset of action and significant side effects. Plasmapheresis and intravenous immunoglobulin have been used mostly for the treatment of severe exacerbations. In the present study we examined the use of intravenous immunoglobulin as maintenance treatment in MG. We included 11 patients with generalized myasthenia gravis. All had severe bulbar and respiratory involvement that required mechanical ventilation in three patients. Intravenous immunoglobulin treatment was initiated at a dose of 400 mg/kg/d for 5 days and followed by maintenance with 400 mg/kg once monthly. Regular medications were continued as necessary. There was significant improvement in all patients, and none required mechanical ventilation over the treatment period of 20.3 months +/- 8.3 (mean +/- SD, total patient years of treatment = 18.7). Steroid and pyridostigmine doses were reduced significantly and steroids were discontinued in two patients. There were no serious side effects related to intravenous immunoglobulin. These results suggest that intravenous immunoglobulin maintenance therapy is a valid modality in patients with MG. PMID- 11391131 TI - Quick titration of pergolide in cotreatment with domperidone is safe and effective. AB - The purpose of the study was to analyze efficacy and safety of quick pergolide titration combined with domperidone. In an open-label prospective study, pergolide was titrated in 16 days to a maximum of 3 mg/d doses as adjunctive treatment to L-Dopa in 10 elderly patients with Parkinson's disease. Sixty milligrams domperidone was started 2 days before and and continued during the pergolide titration period to prevent side effects. Adverse events were studied for 6 weeks. Efficacy was measured with the motor part ("on" condition) of the Unified Parkinson's Disease Rating Scale (UPDRS), the 2-minute walking test, the Timed Up and Go test, and the Postural-Locomotor-Manual test. After quick titration of pergolide with domperidone cotreatment, no symptomatic side effects were seen except for lightheadedness in one patient, which disappeared after dose reduction. The UPDRS motor score improved significantly from 21 +/- 8 at baseline to 16 +/- 7 and 12 +/- 7 after 1 and 2 weeks, respectively. The 2-minute walking distance improved significantly from 123 +/- 36 m at baseline to 136 +/- 41 m after 6 weeks. The Timed Up and Go and Postural-Locomotor-Manual test results, overall, did not show significant changes. Quick titration of pergolide to a maximum of 3 mg/d with domperidone cotreatment is safe and effective. Therefore, we recommend domperidone cotreatment in the titration period to prevent unnecessary failure of dopamine agonist treatment because of adverse effects. PMID- 11391133 TI - Consensus statement on submission and publication of manuscripts. PMID- 11391132 TI - Sleep attacks and antiparkinsonian drugs: a pilot prospective pharmacoepidemiologic study. AB - A prospective survey was performed to characterize the prevalence of sleep attacks and to evaluate precipitating factors in a group of 236 patients with idiopathic Parkinson's disease. Sleep attacks were reported by 72 patients (30.5%). Multivariate analysis showed a marked association between the occurrence of sudden sleep episodes and first autonomic failure, followed by treatment with ropinirole and bromocriptine. The present work underlines the major contributing role of autonomic failure followed by dopamine agonists in the occurrence of such an event. Because a relationship between sleep attacks and not only ropinirole but also bromocriptine treatment was described, the present work suggests that sleep attacks are a common side effect of all dopamine agonists. PMID- 11391134 TI - Long-term results of ileal pouch-anal anastomosis for colorectal Crohn's disease. AB - INTRODUCTION: The aim of this study is to report ten-year results of ileal pouch anal anastomosis in selected patients with colorectal Crohn's disease for whom coloproctectomy and definitive end ileostomy was the only alternative. METHODS: 41 patients (22 females/19 males) with a mean age of 36 +/- 13 (range, 16-72) years underwent ileal pouch-anal anastomosis for colorectal Crohn's disease between 1985 to 1998. None had past or present history of anal manifestations or evidence of small-bowel involvement. Diagnosis of Crohn's disease was established preoperatively in 26 patients, on the resected specimen after ileal pouch-anal anastomosis, or after occurrence of Crohn's disease-related complication in 15 patients. RESULTS: Follow-up was 113 +/- 37 months, (18-174) 20 patients having been followed for more than 10 years. There was no postoperative death. Eleven (27 percent) patients experienced Crohn's disease-related complications, 47 +/- 34 months (8-101) after ileal pouch-anal anastomosis: 2 had persistent anal ulcerations with pouchitis and granulomas on pouch biopsy and were treated medically; 2 experienced extrasphincteric abscesses and 7 presented pouch perineal fistulas which were treated surgically. Among them, 3 patients with persistent perineal fistula despite surgery required definitive end-ileostomy. Of the 20 patients followed for more than 10 years, 7 (35 percent) experienced Crohn's disease-related complications which required pouch excision in 2 (10 percent). CONCLUSIONS: Ten years after ileal pouch-anal anastomosis for colorectal Crohn's disease, rates of Crohn's disease-related complications and pouch excision were 35 and 10 percent, respectively. These good long-term results justify for us to propose ileal pouch-anal anastomosis in selected patients with colorectal Crohn's disease (i.e., no past or present history of anal manifestations and no evidence of small-bowel involvement) for whom the only alternative is definitive end ileostomy. PMID- 11391135 TI - Effect of sacral nerve stimulation in patients with fecal and urinary incontinence. AB - PURPOSE: Preliminary studies have shown improvement in fecal incontinence in several patients who received temporary or permanent stimulation. The purpose of this study was to report our experience in sacral nerve stimulation in the treatment of fecal incontinence and to target patients who would benefit most from stimulation. METHODS: Patients with fecal incontinence were studied clinically and manometrically before, during, and after temporary nerve stimulation. If temporary nerve stimulation was clinically successful, the patient was implanted and followed up for six months. RESULTS: Nine patients (6 female) with a mean age of 50.7 +/- 12.3 years underwent temporary nerve stimulation. Temporary nerve stimulation was successful in eight patients, six of whom were implanted. Of the patients who could be evaluated, three of five had improved at the six-month follow-up visit, particularly in relation to the number of urgency episodes and delay in postponing defecation. All implanted patients had urinary symptoms. Urinary urgency was also improved by stimulation. During temporary nerve stimulation, the maximal squeeze pressure amplitude increased. After implantation, only the duration of maximal squeeze pressure seemed to improve. CONCLUSION: Sacral nerve stimulation can be used in the management of fecal incontinence, particularly in cases of urge fecal incontinence associated with urinary urgency. This study seems to confirm the effect of sacral nerve stimulation on striated sphincter function. PMID- 11391137 TI - Is there an association between fecal incontinence and lower urinary dysfunction? AB - BACKGROUND: Urinary and fecal incontinence in females are both common and distressing conditions. Because common pathophysiologic mechanisms have been described, an association between the two would be expected. The aim of this study was to determine whether patients with lower urinary tract dysfunction have concomitant fecal incontinence when compared with age and gender matched community controls and, second, to determine whether they have predisposing factors that have led to lower urinary tract symptoms and concomitant fecal incontinence. METHODS: A case-control study was performed by means of detailed questionnaire and review of investigation results. One thousand consecutive females presenting for urodynamic investigation of lower urinary tract dysfunction, were compared with 148 age and gender matched community controls. RESULTS: Frequent fecal incontinence was significantly more prevalent among all cases than among community controls (5 vs. 0.72 percent, P = 0.023). Occasional fecal incontinence was also more prevalent (24.6 vs. 8.4 percent, P < 0.001). Fecal incontinence was not significantly more prevalent among females with genuine stress incontinence (5.1 percent) when compared with females with detrusor instability (3.8 percent) or any other urodynamic diagnosis. Symptoms of fecal urgency and fecal urge incontinence were significantly more prevalent among those with a urodynamic diagnosis of detrusor instability or sensory urgency than among females with other urodynamic diagnoses or community controls. Multivariate analysis comparing cases with fecal incontinence with other cases and also with community controls did not indicate that individual obstetric factors contributed significantly to the occurrence of fecal incontinence in these patients. CONCLUSIONS: There is an association between genuine stress incontinence, lower urinary tract dysfunction, and symptoms of fecal incontinence, but the exact mechanism of injury related to childbirth trauma is questioned. PMID- 11391139 TI - Preliminary results of an outcome tool used for evaluation of surgical treatment for fecal incontinence. AB - PURPOSE: The lack of an outcome tool to evaluate the outcome of surgical and medical treatment for fecal incontinence makes interpretation of success difficult. The purpose of this study was to evaluate a preliminary outcome tool for fecal incontinence. METHOD: Since 1994 an extensive database has prospectively been collected on all females undergoing an overlapping sphincter repair for fecal incontinence by a single surgeon. A simple incontinence form designed to examine outcome, developed by colon and rectal surgeons, was filled out preoperatively and postoperatively. RESULTS: Of 206 females evaluated for surgical treatment of their fecal incontinence, 65 underwent surgical treatment from January 1994 until July 1999. The mean age was 49 (range, 23-80) years, and the mean follow-up was 10 (range, 1-50) months. When comparing each variable (problems holding gas, staining of undergarments, accidental bowel movements, and need to wear pads) and lifestyle issue (physical, social, and sexual activities) preoperatively and postoperatively, there was significant improvement in all areas. Three parameters were chosen (change in accidental bowel movements, improvement in two of three lifestyle areas, and improvement in one of three lifestyle areas) to examine individual items from the database and to determine if they affected outcome. No single variable has a significant effect on the outcome. A scoring system was devised from the questionnaire. From preoperatively to postoperatively, there was a median 14-point improvement that was statistically significant. CONCLUSIONS: This preliminary tool to examine outcome for fecal incontinence measures parameters that are significantly improved by overlapping sphincteroplasty. More work is needed to refine and validate this tool because a standard outcome tool is needed for reporting the results of surgical treatment of fecal incontinence. PMID- 11391140 TI - Abdominosacral resection for primary irresectable and locally recurrent rectal cancer. AB - PURPOSE: The purpose of this study was to present a technique of abdominosacral resection and its results in patients with locally advanced primary or locally recurrent rectal cancer with dorsolateral fixation. METHODS: Between 1994 and 1999, 13 patients with locally advanced primary rectal cancer and 37 patients with locally recurrent rectal cancer underwent abdominosacral resection as part of a multimodality treatment, i.e., preoperative irradiation, surgery, and intraoperative irradiation. After the abdominal phase, the patient was turned from supine to prone position to perform the transsacral phase of the resection. RESULTS: Margins were microscopically negative in 26 patients (52 percent), microscopically positive in 18 (36 percent), and positive with gross residual disease in 6 patients. Operation time ranged from 210 to 590 (median, 390) minutes, and blood loss ranged from 400 to 10,000 (median, 3,500) ml. No operative or hospital deaths occurred. Postoperative complications occurred in 41 patients (82 percent); most notable were perineal wound infections or dehiscence (n = 24, 48 percent). Other complications were postoperative urinary retention or incontinence (n = 9, 18 percent), peritonitis (n = 4), grade II neuropathy (n = 1), and fistula formation (n = 3). Kaplan-Meier 3-year overall survival, disease free survival, and local control rates were, respectively, 41 percent, 31 percent, and 61 percent. Completeness of the resection (negative vs. positive margins) was a significant factor influencing survival (P = 0.04), disease-free survival (P = 0.0006), and local control (P = 0.0002). CONCLUSION: The abdominosacral resection provides wide access and may be the therapeutic solution for the accomplishment of a radical resection for distally situated, dorsally or dorsolaterally fixed primary or locally recurrent rectal cancers. PMID- 11391141 TI - Stratifying risk factors for follow-up: a comparison of recurrent and nonrecurrent colorectal cancer. AB - INTRODUCTION: The selection of patients for individualized follow-up and adjuvant therapy after curative resection of colorectal carcinoma depends on finding reliable prognostic criteria for recurrence. However, such criteria are not universally accepted, and follow-up is often standardized for all patients without regard for each individual's level of risk of recurrence. Such a system of follow-up is not cost-effective. METHODS: A comparison of operative findings, pathologic features, and follow-up data of 1,731 cases of nonrecurrent colorectal cancer (821 colon, 910 rectum) with 357 cases of recurrent colorectal cancer (164 colon, 193 rectum) following potentially curative surgery was made, and results were analyzed to ascertain criteria for stratifying follow-up according to risk factors. RESULTS: Single-factor analysis showed that Dukes staging and tumor invasion were significantly associated with recurrence in both rectal and colon carcinoma. Tumor fixation and grading were additional significant factors in rectal cancer. Recurrence rates, time to recurrence, site of recurrence (locoregional vs. distant), and pattern of metastatic spread were not significantly affected by original tumor site. Recurrence was not significantly affected by patient age and gender. Individual surgeon performance in this series had also no significant effects on tumor recurrence. With multivariate analysis only, Dukes staging and tumor invasion into adjacent tissues were found to be independent adverse prognostic factors for recurrence. CONCLUSIONS: Dukes staging and tumor penetration into adjacent tissues are the only significant adverse prognostic factors for tumor recurrence of colonic and rectal carcinoma. Tumor grade and tumor fixation are additional adverse prognostic factors in rectal cancer. Guidelines for follow-up may be based on these factors and follow-up thus stratified according to risk of developing recurrence. PMID- 11391142 TI - Postoperative adhesions: ten-year follow-up of 12,584 patients undergoing lower abdominal surgery. AB - PURPOSE: Postoperative adhesions are a significant problem after colorectal surgery. However, the basic epidemiology and clinical burden are unknown. The Surgical and Clinical Adhesions Research Study has investigated the scale of the problem in a population of 5 million. METHODS: Validated data from the Scottish National Health Service Medical Record Linkage Database were used to define a cohort of 12,584 patients undergoing open lower abdominal surgery in 1986. Readmissions for potential adhesion-related disease in the subsequent ten years were analyzed. The methodology was conservative in interpreting adhesion-related disease. RESULTS: In the study cohort 32.6 percent of patients were readmitted a mean of 2.2 times in the subsequent ten years for a potential adhesion-related problem. Although 25.4 percent of readmissions were in the first postoperative year, they continued steadily throughout the study period. After open lower abdominal surgery 7.3 percent (643) of readmissions (8,861) were directly related to adhesions. This varied according to operation site: colon (7.1 percent), rectum (8.8 percent), and small intestine (7.6 percent). The readmission rate was assessed to provide an indicator of relative risk of adhesion-related problems after initial surgery. The overall average rate of readmissions was 70.4 per 100 initial operations, with 5.1 directly related to adhesions. This rose to 116.4 and 116.5, respectively, after colonic or rectal surgery-with 8.2 and 10.3 directly related to adhesions. CONCLUSIONS: There is a high relative risk of adhesion-related problems after open lower abdominal surgery and a correspondingly high workload associated with these readmissions. This is influenced by the initial site of surgery, colon and rectum having both the greatest impact on workload and highest relative risk of directly adhesion related problems. The study provides sound justification for improved adhesion prevention strategies. PMID- 11391143 TI - Impotence after mesorectal and close rectal dissection for inflammatory bowel disease. AB - PURPOSE: Close rectal dissection is a surgical technique used by some surgeons in inflammatory bowel disease. It is performed within the mesorectum, close to the rectal muscle wall, with the aim of minimizing damage to the pelvic sexual nerves. Other surgeons dissect in the more anatomical mesorectal plane. Our aim was to determine whether close rectal dissection is more protective of the pelvic sexual nerves than mesorectal dissection. METHOD: Patients undergoing surgery for inflammatory bowel disease were entered prospectively into a database. Male patients were mailed a standardized, validated, urologic impotence questionnaire: the International Index of Erectile Function. RESULTS: There was an 81 percent response rate. Six of 156 assessable patients were totally impotent (3.8 percent). They were all in the 50-year-old to 70-year-old age group, with no impotence in patients younger than 50 years old. Twenty-one patients complained of minor diminution of erectile function (13.5 percent), where sexual activity was still possible. There was no statistical difference in the rate of complete (2.2 percent vs. 4.5 percent, P = 0.67) or partial (13.5 percent vs. 13.3 percent, P = 0.99) impotence between close rectal and mesorectal dissection (Fisher's exact test). There were no ejaculatory difficulties. The time elapsed since surgery ranged from 2.7 months to 192.7 months, with a median of 74.5 months. CONCLUSION: Rectal excision for inflammatory bowel disease can be conducted with low rates of impotence. Minor degrees of erectile dysfunction may be more common than currently recognized. We could not demonstrate that close rectal dissection significantly protects the patient from impotence compared with operating in the anatomical mesorectal plane. Age appears to be the most important risk factor for postoperative impotence. PMID- 11391144 TI - Early and late (ten years) experience with circular stapler hemorrhoidectomy. AB - PURPOSE: We present a retrospective clinical study concerning the preliminary experience with the circular stapler in the treatment of hemorrhoids. Early results, complications, and long-term follow-up are revisited. METHODS: Fifty-six consecutive patients with second-, third-, and fourth-degree hemorrhoids were included in the study. Data about operation, early postoperative results, and follow-up at one, two, and four weeks were collected. Patients were also contacted by phone after a long-term follow-up (mean, 33 (range, 5-120) months). RESULTS: Every operation attempted was successfully terminated. The length of the operation was less than 15 minutes. No major bleeding or anastomotic disruption occurred. Six patients (13 percent) who underwent spinal or epidural anesthesia had urinary retention. One patient (1.7 percent) had minor bleeding, and four patients (7.1 percent) experienced transient edema of the anastomotic ring after the operation. None needed further treatments. The mean analgesic requirement was 1.4 (range, zero to eight) ketorolac 30-mg injections; 23 patients (41 percent) received no analgesics, and seven patients (12 percent) required a single extra dose of opiates (10 mg morphine cloridrate). Length of hospital stay was between 0 and 11 (mean, 2.7) days, but 20 patients (35 percent) received an additional operation for coexisting surgical disease. At one week, almost all patients experienced little pain at digital inspection and little bleeding after defecations. No anastomotic leakage, wound infection, or healing delay was found. Three patients (5.3 percent) experienced wound edema and pain during defecation. Two weeks later, one patient (1.7 percent) suffered from painful defecation and ten patients (17 percent) reported minor bleeding, but all returned to normal activities. No pain during defecation, bleeding, stenosis, soiling, incontinence, or other anal symptoms were found at one month after the operation, and all patients were well. All patients were contacted by phone 5 to 120 (mean, 33) months later, and all were pleased with the results of this procedure. There were no symptomatic recurrences. DISCUSSION: Our study confirms the feasibility of circular stapler hemorrhoidectomy in the treatment of hemorrhoids. Complications and postoperative pain were minimal. There were no recurrences during long-term follow-up. CONCLUSION: Mechanical hemorrhoidectomy is a promising new option in the treatment of all patients eligible for a surgical approach. PMID- 11391145 TI - Day surgery for mucosal-hemorrhoidal prolapse using a circular stapler and modified regional anesthesia. AB - PURPOSE: In 1993, prolapse reduction using the circular stapler for the treatment of hemorrhoidal disease was proposed. The procedure is characterized by minimal postoperative pain. In this study we evaluated the above technique using regional anesthesia to identify the advantages and feasibility of stapled hemorrhoidectomy, with special focus on the efficacy of same-day discharge. METHODS: From December 1997 to November 1999, we performed 70 consecutive reduction corrections of mucosal hemorrhoidal prolapse using the circular stapler with regional anesthesia (a technical modification of Marti's posterior perineal block). Our series included 41 males and 29 females with a mean age of 43.4 (range, 25-74) years. Three patients were affected by second-degree hemorrhoids and 67 by third-degree hemorrhoids. RESULTS: Sixty-two patients were discharged three hours after the operation in good general condition and without pain, whereas eight patients were discharged the day after for early complications, consisting of two cases of early bleeding, three cases of urinary retention, and three cases of persistent severe pain requiring prolonged medical treatment. CONCLUSION: Our study shows that, in selected cases, it is possible to perform day surgery for patients with hemorrhoidal disease using a circular stapler device when combined with regional anesthesia. PMID- 11391146 TI - Surgical treatment of hemorrhoids: prospective, randomized trial comparing closed excisional hemorrhoidectomy and the Harmonic Scalpel technique of excisional hemorrhoidectomy. AB - PURPOSE: The object of this study was to evaluate technique using the ultrasonically activated scalpel as an alternative to closed hemorrhoidectomy in an unbiased evaluation of this new technology. METHODS: Thirty patients with Grade 2 or 3 symptomatic hemorrhoids were prospectively randomized to undergo closed hemorrhoidectomy assisted by electrocautery or hemorrhoidectomy with the ultrasonically activated scalpel, i.e., the Harmonic Scalpel. We evaluated the difference between techniques in operative time, postoperative pain, incontinence, and quality of life (using the Short Form-36 survey), as well as complications. RESULTS: Mean operative time for closed hemorrhoidectomy with electrocautery was 35.7 +/- 3 minutes; for Harmonic Scalpel patients, it was 31.7 +/- 2 minutes (P < 0.37). There was no statistical difference in operative time for two- or three-column hemorrhoidectomy. There was no significant difference in pain measurements reported on Day 1 (5.8 +/- 0.4 for electrocautery and 5.6 +/- 0.6 for Harmonic Scalpel, P < 0.82). On postoperative Day 7, the difference in pain between groups approached significance, with pain reported as 3.7 +/- 0.3 for electrocautery and 5.1 +/- 0.7 for Harmonic Scalpel(R) (P < 0.06). At six weeks, both groups were pain free. There was a significant decrease in pain between postoperative Days 1 and 7 in the electrocautery patients that was not seen in the Harmonic Scalpel patients. Incontinence measured preoperatively, at postoperative Day 7, and at postoperative Week 6 was similar for both groups and reflected occasional incontinence of gas. When the various items of the Short Form-36 survey were compared, there was no significant difference between posttreatment and preoperative values. There was no difference in the number of complications between patient groups. CONCLUSION: Although the Harmonic Scalpel is an effective tool in the treatment of hemorrhoidal disease, we found no specific advantage in postoperative pain, fecal incontinence, operative time, quality of life, or complications compared with traditional closed hemorrhoidectomy. PMID- 11391147 TI - The role of sentinel lymph node mapping in staging of colon and rectal cancer. AB - PURPOSE: Nodal metastasis is the best predictor of survival for patients with colon cancer. Statistical models based on random distribution of positive lymph nodes suggest that to correctly classify nodal status with 95 percent confidence, 20 nodes are needed for T1 lesions, 17 nodes for T2, and 15 nodes for T3. The mean number of nodes identified in American patients is 8, suggesting that they might not be accurately staged. Patients in our tumor registry staged as "node negative" had a short survival when they had < or =10 lymph nodes evaluated when compared with patients with >10 lymph nodes evaluated (p < 0.01). We hypothesized that the use of sentinel lymph node may assist in the staging of colon cancer. METHODS: Thirty-eight consecutive patients with colon lesions were prospectively enrolled into this trial between February 1998 and November 1999. Thirty-one patients met criteria for analysis. During surgery, Lymphazurin blue dye was injected subserosally into the area around the tumor. Routine nodal evaluation, with extra cuts of all sentinel nodes, was undertaken. RESULTS: At least one sentinel lymph node was found in 18 of 31 patients (58 percent). Sensitivity of 67 percent, specificity and positive predictive value of 100 percent, and negative predictive value of 94 percent were found when sentinel lymph nodes were identified. In 2 of these 18 patients, the sentinel lymph node was the only positive lymph node found. CONCLUSIONS: Application of the sentinel lymph node technique to colon cancer may make it easier to identify lymph nodes most likely to contain metastatic disease, potentially "down-staging" more patients. This may have implications in postoperative care. PMID- 11391148 TI - Total pelvic mesh repair: a ten-year experience. AB - INTRODUCTION: This report describes our technique and experience in restoring the pelvic floor of females with pelvic organ prolapse. METHODS: Total pelvic mesh repair uses a strip of Marlex Mesh secured between the perineal body and the sacrum. Two additional strips, attached to the first, are tunneled laterally to the pubis and support the vagina and bladder laterally. Candidates for the procedure have failed previous standard repair or manifest combined organ prolapse on physical and cystodefecography exams. RESULTS: From January 1990 to December 1999, 236 females had total pelvic mesh repair, and 205 (87 percent) were available for follow-up. Median age was 64 (range, 32-89) years, median parity 2 (range, 1-9); 63 percent had birth-related complications. Bladder protrusion, vaginal protrusion, or both were the predominant chief complaint (54 percent), followed by anorectal protrusion (48 percent). Findings on physical examination showed degrees of prolapse of rectum (74 percent) and vagina (57 percent), perineal descent (63 percent), enterocele (47 percent), and rectocele (44 percent). Mean procedure time and length of hospital stay were 3.2 (standard deviation 0.75) hours and 6 (standard deviation 2.2) days, respectively. Reoperation rate because of complications of the total pelvic mesh repair procedure was 10 percent. Marlex erosion into rectum or vagina occurred in 5 percent of patients and constituted 46 percent of the complications requiring reoperation. Additional surgical procedures at various intervals subsequent to total pelvic mesh repair have been performed in 36 percent of patients to further improve bladder function and have been performed in 28 percent of patients to improve anorectal function. There has been no recurrence of rectal or vaginal prolapse to date. Reports of overall satisfaction for correction of primary symptoms for patients grouped into early (0.5-3 years), middle (>3-6 years) and late (>6 years) were 68 percent, 73 percent, and 74 percent respectively. CONCLUSION: Total pelvic mesh repair is a safe and effective operation for females with pelvic organ prolapse. PMID- 11391149 TI - Reasons for failure of glyceryl trinitrate treatment of chronic fissure-in-ano: a multivariate analysis. AB - PURPOSE: Although glyceryl trinitrate ointment has become the first-line treatment for chronic anal fissure, healing rates are lower than after lateral internal sphincterotomy. The purpose of this study was to identify which factors are associated with treatment failure of glyceryl trinitrate ointment. METHODS: All patients who presented with chronic anal fissure from March 1997 to November 1998 were treated with 0.2 percent glyceryl trinitrate ointment. They were prospectively evaluated until healing or lateral internal sphincterotomy occurred, and long-term follow-up was obtained by standardized telephone questionnaire. A Cox model multivariate analysis was used with seven variables to determine significant factors related to healing. RESULTS: Sixty-four patients (42 men and 22 women; mean age, 37.5 years) with chronic fissure-in-ano were treated with 0.2 percent glyceryl trinitrate ointment. Sentinel piles were observed in 19 patients (29.7 percent). Twenty-six patients (40.6 percent) were healed initially, but 12 (46.2 percent) experienced recurrence. Mean follow-up time was 15.6 (+/-5.5) months. Twenty-nine patients (45.3 percent) had known risk factors for anal fissure, including constipation (n = 21; 32.8 percent), recent childbirth (n = 6; 9.3 percent), colonoscopy (n = 1; 1.6 percent), and anoreceptive intercourse (n = 1; 1.6 percent). Fissures were significantly less likely to heal initially (P < 0.05), more likely to recur (P < 0.05), and more likely to remain unhealed in the long term (P < 0.05) in the presence of a sentinel pile. Fissures with a history of more than six months were less likely to heal initially (P < 0.05). CONCLUSION: The presence of a sentinel pile adversely affects the outcome of treatment of chronic anal fissures with glyceryl trinitrate ointment, and a long history of the fissure reduces the rate of initial healing. Reasons for these findings are discussed. PMID- 11391150 TI - Staged excision and split-thickness skin graft for circumferential perianal Paget's disease. AB - PURPOSE: This study was designed to describe a surgical technique for the treatment of circumferential Paget's disease. METHODS: A search of our computerized patient registry was undertaken, and case records for those patients with perianal Paget's disease were systematically reviewed. RESULTS: Between July 1993 and October 1998, four patients with perianal Paget's disease were identified. Circumferential lesions were identified in three of the four patients. Wide local excision was considered the procedure of choice. All patients underwent a two-staged excision using split-thickness skin graft reconstruction. A similar second stage was performed six to eight weeks later; the other half of the circumference was excised and grafted. No patient had a protective stoma. Graft survival was 100 percent for two patients (four operations) and 80 percent and 70 percent for the other two patients (two operations). The remaining surface healed successfully by secondary intention. In one patient, residual disease was positive at one margin, and a third local excision and split-thickness skin graft was performed. Hospital stay ranged from five to nine days for each procedure. There were no major complications; one patient developed a mild anal stenosis three months after the second procedure and was successfully medically treated. CONCLUSIONS: Staged excision and split thickness skin graft is a viable option for the treatment of circumferential perianal lesions. It carries a minimal morbidity and no observed mortality, the functional result is good, and it is technically simple compared with myocutaneous grafts. Moreover, a stoma is not required. PMID- 11391151 TI - Does stool collection method affect outcomes in immunochemical fecal occult blood testing? AB - PURPOSE: This paper compares the positive predictive value of an immunochemical fecal occult blood test for colorectal neoplasms between the stool specimens obtained during the digital rectal examination and those obtained during the routine screening. METHODS: In a medical check-up, 1,688 subjects received both an immunochemical fecal occult blood test and a colonoscopy. Fecal occult blood was tested by two methods: digital rectal examination and routine screening. The positivity rate of an immunochemical fecal occult blood test and the positive predictive value for colorectal cancer and large adenomatous polyp were determined by these two methods. RESULTS: The positivity rate and the positive predictive value were 5.4 percent and 19.8 percent (4.4 percent for cancer and 15.4 percent for adenomatous polyp) in the digital rectal examination method and 3.5 percent and 27.1 percent (6.8 percent for cancer and 20.3 percent for adenomatous polyp) in the routine screening method, respectively. These figures indicate a significant difference in the positivity rate (P < 0.01) and the positive predictive value (P < 0.05) between these two methods. CONCLUSIONS: These findings indicate that the stool obtained by routine screening has a better positive predictive value than stool collected during the digital rectal examination. PMID- 11391152 TI - The role of tumor cell adhesion as an important factor in formation of distant colorectal metastasis. AB - PURPOSE: The interactions of blood-borne colorectal carcinoma cells with vascular endothelium are important during hematogenous formation of distant metastases. To adhere to the vessel wall, circulating carcinoma cells that come into contact with the microvasculature must resist the attractive forces of the flow of plasma and other circulating cells that tend to detach them from the wall. METHODS: Hydrodynamic adhesion assays have been introduced to mimic the microcirculation and investigate cell adhesion under flow conditions. Different aspects of colorectal cancer cell adhesion during hematogenous formation of distant metastases are summarized and discussed in this review. RESULTS: Adhesion of colorectal carcinoma cells to endothelial cells and extracellular matrix is influenced by the presence of fluid flow. Shear forces alone are able to induce signal transduction events in these cells that result in cell activation and modification of adhesive behavior. CONCLUSIONS: Consideration of fluid dynamics of circulating colorectal cancer cell movement in the microcirculation leads to new knowledge of in vivo processes that are involved in tumor cell adhesion to the vessel wall in host organs. Shear forces have been found to influence adhesive properties of colorectal carcinoma cells to endothelial cells and underlying subendothelial extracellular matrix. Understanding the complex processes involved in tumor cell adhesion may result in the development of novel therapeutic strategies. PMID- 11391153 TI - Mesenteric panniculitis of the colon with obstruction of the inferior mesenteric vein: report of a case. AB - Mesenteric panniculitis is a rare disease characterized by nonspecific inflammation of the fat tissue of the mesentery. We present an extremely rare case of mesenteric panniculitis of the sigmoid colon, complicated by occlusion of the inferior mesenteric vein. A 75-year-old male presented with a one-month history of abdominal distention and abdominal mass without pain. Physical examination revealed a firm mass in the lower abdomen. Barium enema study demonstrated rugged mucosa and a serrated contour in the rectosigmoid colon. Computed tomography showed that the mass arose from the mesentery, which surrounded the mesenteric vessels. The density of the mass was slightly higher than that of fatty tissue. Based on these radiologic findings, the patient was diagnosed as having mesenteric panniculitis of the rectosigmoid colon. Colonoscopy showed narrowing with edematous mucosa in the rectosigmoid colon, whereas marked dilated vessels were noted in the proximal portion of the sigmoid colon. Angiography showed occlusion of the inferior mesenteric vein, with venous flow returning via a collateral vein. The patient was observed without medication because his condition was satisfactory. His symptoms subsequently disappeared during a period of several weeks. The mass in the lower abdomen gradually diminished in size, disappearing three months later. Computed tomography and barium enema showed improvement of the lesion. The favorable outcome of the present case was probably because of formation of a collateral vein. The present case suggests that aggressive therapy for mesenteric panniculitis should be avoided, because the outcome of this disorder is good, even when there is obstruction of vessels. PMID- 11391154 TI - Rectal endometrial stromal sarcoma arising in endometriosis: report of a case. AB - PURPOSE: Endometriosis of the rectovaginal septum can harbor different types of secondary tumors that may involve the rectal wall and protrude into its lumen, thus making diagnosis difficult. Extrauterine low-grade endometrial stromal sarcoma may rarely arise in endometriosis. The purpose of this article was to present the third case of this association. METHOD: This was a clinicopathologic study. RESULTS: A 42-year-old female presented with abdominal pain and fever. Laparotomy revealed a large pelvic mass involving the rectovaginal septum and the colonic wall and which protruded into the lumen forming endoluminal polypoid masses. Concomitant peritoneal nodules and a metastatic paracolic lymph node were also found. Histopathologically, primary endometriotic foci were found in close relationship with an endometrial stromal sarcoma which invaded the rectal wall. The female genital tract had no endometriotic lesions. The patient was treated by surgery and subsequent chemotherapy and was alive and well 20 months later. CONCLUSIONS: Endometriosis and its possible malignant changes should be taken into account in the differential endoscopic diagnosis of rectal masses in females. PMID- 11391155 TI - Cecal volvulus causing postoperative intestinal obstruction: report of a case. AB - Cecal volvulus is a rare cause of intestinal obstruction after major abdominal surgery. A case of cecal volvulus occurring in the early postoperative period after left colon resection for malignancy is presented. Clinical evaluation and plain abdominal radiographs suggesting cecal volvulus prompted laparotomy and correction. Delay in diagnosis results in high mortality, and treatment depends largely on the viability of the involved intestine. This report describes the second case of cecal volvulus complicating a left colectomy. It was treated by detorsion and reperitonealization cecopexy. PMID- 11391156 TI - Gasless hand-assisted laparoscopic surgery for colorectal cancer: an option for poor cardiopulmonary reserve. AB - Gasless hand-assisted surgery, an alternative technique for colorectal cancer, is described. The abdomen is lifted by two metal disks especially designed for this procedure. The gasless condition evokes no hazard from pneumoperitoneum, and the procedure is greatly simplified by the hand assistance. PMID- 11391157 TI - Practice parameters for antibiotic prophylaxis to prevent infective endocarditis or infected prosthesis during colon and rectal endoscopy. PMID- 11391158 TI - Quantification of in vivo replicative capacity of HIV-1 in different compartments of infected cells. AB - Based on a mathematical model, we analyze the dynamics of CD4+ cells, actively, latently, persistently, and defectively infected cells and plasma virus after initiation of antiretroviral therapy in 14 HIV-1-infected asymptomatic patients. By simultaneous fitting of our model to clinical data of plasma HIV-1 RNA, peripheral blood mononuclear cell (PBMC)-gag RNA, proviral DNA, and CD4+ cell counts, we estimate kinetic parameters to determine the basic reproductive rate (R0) of the virus in different infected cell compartments as a measure of the replicative capacity of the virus in vivo. We find that the basic reproductive rate is larger than 1 before treatment only in actively infected cells (mean R0(act) approximately 2.46) indicating that only in this compartment the virus can maintain an ongoing infection. In latently and persistently infected cells the basic reproductive rate is considerably smaller (R0(lat) approximately 0.03 and R0(pers) approximately 0.008, respectively) indicating that these compartments contribute little to the total basic reproductive rate and cannot maintain an ongoing infection in absence of actively infected cells. PMID- 11391159 TI - Effect of drug efficacy and the eclipse phase of the viral life cycle on estimates of HIV viral dynamic parameters. AB - Fits of mathematic models to the decline in HIV-1 RNA after antiretroviral therapies have yielded estimates for the life span of productively infected cells of 1 to 2 days. In a previous report, we described the mathematic properties of an extended model that accounts for imperfect viral suppression and the eclipse phase of the viral life cycle (the intracellular delay between initial infection and release of progeny virions). In this article, we fit this extended model to detailed data on the decline of plasma HIV-1 RNA after treatment with the protease inhibitor ritonavir. Because the therapy in this study was most likely not completely suppressive, we allowed the drug efficacy parameter to vary from 70% to 100%. Estimates for the clearance rate of free virus, c, increased with the addition of the intracellular delay (as reported previously) but were not appreciably affected by changes in the drug efficacy parameter. By contrast, the estimated death rate of virus-producing cells, delta, increased from an average of 0.49 day-1 to 0.90 day-1 (an increase of 84%) because the drug efficacy parameter was reduced from 100% to 70%. Neglecting the intracellular delay, the comparable increase in delta was only about 55%. The inferred increases in delta doubled when the model was extended to account for possible increases in target cell densities after treatment initiation. This work suggests that estimates for delta may be greater than previously reported and that the half-life of a cell in vivo that is producing virus, on average, may be 1 day. PMID- 11391160 TI - Immunogenicity of recombinant envelope glycoproteins derived from T-cell line adapted isolates or primary HIV isolates: a comparative study using multivalent vaccine approaches. AB - We investigated immunogenic properties of native envelope glycoproteins derived from HIV-1 (subtype B). Our main objective was to assess whether the design of multivalent vaccines affects generation of neutralizing antibodies against primary viruses. Recombinant Semliki Forest virus (SFV) particles producing various HIV-1 envelope glycoproteins were used as vaccine vectors. The following multivalent vaccination approaches were compared: 1) immunization with a mixture of recombinant SFV expressing envelope glycoproteins derived from three HIV-1 primary isolates and two T-cell laboratory-adapted (TCLA) viruses; 2) immunization with a mixture of recombinant SFV expressing only the envelope glycoproteins derived from three HIV-1 primary isolates; 3) sequential immunizations with the recombinant SFV expressing the envelope glycoproteins derived from three HIV-1 primary isolates and two TCLA viruses, respectively. Two monovalent vaccine approaches using SFV expressing envelope glycoproteins derived from a single primary isolate or TCLA virus were also included in the study. The multivalent vaccination strategies based on SFV vaccine vectors did not induce more neutralizing antibodies than the previously tested TCLA envelope immunogens, which gave disappointing results against primary isolates. PMID- 11391161 TI - A pilot study of the use of mycophenolate mofetil as a component of therapy for multidrug-resistant HIV-1 infection. AB - Mycophenolic acid (MPA) increases the activity of both abacavir (ABC) and didanosine (ddI) in vitro against wild-type and multinucleoside-resistant HIV. We treated 7 patients with diagnosed AIDS who did not respond to eight or more antiretroviral therapies in an open label pilot study with mycophenolate mofetil (MMF), ABC, ddI, amprenavir (APV), and ritonavir (RTV), with or without efavirenz (EFV). Therapy was well tolerated despite the patients' advanced disease states. No significant decline in lymphocyte or other blood counts was observed. Median HIV RNA was 5.26 log10 copies/ml at entry, 4.53 log10 copies/ml at 4 weeks, and 5.13 log10 copies/ml at 16 weeks. Median CD4+ count was 34 cells/microl at entry and 39 cells/microl at 16 weeks of therapy. CD4+ counts increased further in five study subjects on extended therapy to 25 weeks (median 27 cells/microl at entry, 66 cells/microl at close), despite loss of virologic suppression in 4 of 5 cases. MPA can induce apoptosis in lymphocytes in vitro. However despite viral rebound, cell surface markers of apoptosis and activation declined in total CD3+ cells and CD3+/CD4+ cells twofold to fourfold in 4 of 5 adherent study subjects at 16 weeks, reaching levels comparable with those found in seronegative donors. Although low-dose MMF appears safe in late-stage HIV disease, this study did not demonstrate virologic efficacy. Higher doses of MMF may be more effective. With careful monitoring of toxicities and pharmacokinetics, MMF deserves further testing in HIV therapy. PMID- 11391163 TI - Altered fat distribution in HIV-positive men on nucleoside analog reverse transcriptase inhibitor therapy. AB - To determine whether HIV infection, the wasting syndrome, or nucleoside analog reverse transcriptase inhibitor (NRTI) or protease inhibitor (PI) therapy uniquely affect fat distribution in men, we performed manual regional analysis of total, appendicular, trunk, and central abdominal fat measured by dual-energy X ray absorptiometry (DEXA). Five groups of study subjects were identified for this cross-sectional analysis: HIV-negative controls (HIV-; N = 44) and four groups of HIV-positive subjects: antiretroviral (ARV)-naive or with limited prior use of NRTIs (ARV-; N = 23); on NRTIs for > or =6 months but PI-naive (NRTI; N = 30); on an NRTI/PI regimen for > or =6 months but with no complaints of abnormal fat distribution (NRTI/PI; N = 26); and those on NRTIs but PI-naive with the wasting syndrome (NRTI/WS; N = 40). Total, appendicular, trunk, and central abdominal fat was significantly lower in NRTI/WS. The ratio of trunk fat to appendicular fat was virtually identical in HIV- and ARV-. This ratio was significantly higher in the NRTI, NRTI/PI, and NRTI/WS groups, and values in these three groups were similar. These cross-sectional data suggest that HIV-infected men receiving NRTIs have an altered pattern of fat distribution, compared with HIV-negative men and HIV-positive men who are not receiving antiretroviral therapy. This effect was independent of the concomitant use of a PI or a diagnosis of the wasting syndrome. We saw no evidence of a unique effect of HIV infection per se on regional fat distribution. Although the fat ratio is increasingly employed, its physiologic significance is unclear. Our results, which have been obtained retrospectively, are intended to provide the impetus for prospective, controlled studies of the interactions among drug and host factors in the development of fat distribution abnormalities. PMID- 11391162 TI - Provider assessment of adherence to HIV antiretroviral therapy. AB - BACKGROUND: Adherence assessment is an essential component of monitoring HIV antiretroviral therapy. Prior studies suggest that medical providers frequently estimate individual patient adherence inaccurately. OBJECTIVE: We compared provider estimates of nonadherence to antiretroviral therapy with unannounced pill counts and structured patient interviews to determine the accuracy of adherence information obtained by providers and patients. DESIGN, SETTING, AND PARTICIPANTS: Comparison of three adherence measures in homeless or marginally housed persons receiving HIV antiretroviral therapy (n = 45) and their providers (n = 35). MEASUREMENTS: Provider estimate of percentage of pills taken; three successive patient structured reports of number of doses missed in the last 3 days; and three successive unannounced pill counts. RESULTS: 13% (95% confidence interval [CI], 4%-22%) of patients were not following their regimen as directed. Provider-adherence estimate explained only 26% (95% CI, 6%-47%) of the variation in pill count adherence, whereas patient report explained 72% (95% CI, 52%-96%). The sensitivity and specificity of provider estimates of nonadherence, defined as <80% of pills taken by pill count, were 40% and 85%, respectively. The sensitivity and specificity of patient interview were 72% and 95%, respectively. CONCLUSIONS: Provider estimate of adherence was inaccurate whereas structured patient report was more closely related to pill count. Structured assessment over several short intervals may improve accuracy of adherence assessment in clinical practice. PMID- 11391164 TI - HIV-1 RNA levels and development of clinical disease in two different adolescent populations. AB - HIV infection rates in American youth continue to increase unabated. As adolescent-specific therapeutic interventions are planned, information on HIV infection's course and its predictors becomes critically important for valid and precise study design. We report on age-specific disease rates stratified by estimated time since infected and predictors of HIV disease progression through four clinical categories in two distinct adolescent populations. Adolescents with hemophilia infected through contaminated blood products showed disease progression rates of 18 to 23 events per 100 person-years (PYs) by age and years infected. Predictors of first progression included HIV-1 RNA >30,000 copies/ml (rate ratio [RR], 2.4; 95% confidence interval [CI], 1.5-3.9), antiretroviral monotherapy (RR, 2.4; 95% CI, 1.7-3.3); Latino/a ethnicity (RR, 2.2; 95% CI, 1.2 4.2) and initial intermediate clinical status (RR, 1.9; 95% CI, 1.3-2.9). Sexually-infected adolescents >18 years who had been infected >3 to 6 years had a disease progression rate of 16 events per 100 PY. For these youths, the sole predictor of first progression was viral load (VL) (RR for VL >30,000 copies per ml, 8.4; 95% CI, 2.8-25.1). This article examines the predictive capacity of viral load and evaluates other cofactors for disease progression in different adolescent populations. These data will be of value in clinical trial design. PMID- 11391165 TI - A phase II trial of dual protease inhibitor therapy: amprenavir in combination with indinavir, nelfinavir, or saquinavir. AB - This study evaluated dual protease inhibitor (PI) regimens containing amprenavir (APV) in PI-naive, HIV-1-infected patients over 48 weeks. Patients were randomized to 800-mg APV combined with 800-mg indinavir (IDV), 750-mg nelfinavir (NFV), or 800-mg saquinavir-soft gel capsule (SGV-SGC), all three times daily without nucleoside reverse transcriptase inhibitors, or APV given alone for 3 weeks and then with 150-mg lamivudine (3TC) and 300-mg zidovudine (ZDV), twice daily. Dual PI therapy demonstrated substantial antiviral activity and was generally safe and well tolerated. Eight patients had virologic failure; 5 were receiving dual PI therapy and 3 were in the APV/3TC/ZDV arm. The protease I50V mutation characteristic of APV resistance was not observed, although other key PI mutations were selected in 4 patients failing therapy, 2 of whom had PI resistance at baseline. PMID- 11391166 TI - Long-term follow-up of antiretroviral-naive HIV-positive patients treated with nevirapine. AB - This article reports on the extended follow-up of 125 antiretroviral (ARV)-naive patients treated with nevirapine (NVP) in the United Kingdom. The patients have been observed for a median of 1.8 years after starting NVP (range, 4 days-2.7 years). Baseline CD4 counts and HIV RNA levels were 210 (interquartile range, 130 - 335) cells/mm3 and 4.86 (range, 4.52-5.26) log10 copies/ml, respectively. Eleven patients (9.0%) developed a rash thought to be related to NVP, of whom 4 permanently discontinued NVP. Twenty-four months after starting NVP, RNA levels had dropped by a median of 2.32 log10 copies/ml and CD4 counts increased by a median of 143 cells/mm3. In all, 96 patients had at least one viral load measured <500 copies/ml, a median of 2.8 months after starting NVP. RNA levels rebounded >500 copies/ml in 37 of these patients, on average 2 years after initial response. In conclusion, in ARV-naive patients, NVP is generally well tolerated and long-term response rates are good. PMID- 11391167 TI - Relation between HIV-1 and hepatitis C viral load in patients with hemophilia. AB - Coinfection with hepatitis C virus (HCV) and HIV-1 is common in patients with hemophilia and in intravenous drug users. Little, however, is known about the relation between HIV-1 and HCV coinfection and the effects on HCV clearance and pathogenesis. We examined data from 207 HIV-1-infected and 126 HIV-1-uninfected patients with hemophilia enrolled in the multicenter Hemophilia Growth and Development Study. Participants were observed during prospective follow-up for approximately 7 years with annual measurements of alanine aminotransferase (ALT), CD4+ cells, and HCV and HIV-1 RNA levels. Clearance of HCV was more likely to occur in those uninfected with HIV-1 (14.3 versus 2.5%; odds ratio [OR] 4.79; 95% confidence interval [CI], 1.63-14.08, p =.005) and was more common with decreasing age (OR, 1.23; 95% CI, 1.04-1.47; p =.017). HCV RNA levels were higher throughout the 7 years of follow-up in those HIV-1-infected (p <.001). In the HIV 1-infected participants, baseline CD4+ cells were inversely related to HCV RNA with every 100-cell increase associated with a 0.19 log10 copy/ml decrease in HCV RNA (p =.002), and HIV-1 and HCV RNA levels were directly related (p =.008). Increasing HCV RNA levels were also associated with significantly higher ALT levels regardless of HIV-1 infection status. These results demonstrate that HIV 1/HCV co-infection is associated with a reduced likelihood of HCV clearance and that higher levels of HCV RNA are associated with increased hepatic inflammation. PMID- 11391168 TI - Response to highly active antiretroviral therapy according to duration of HIV infection. AB - OBJECTIVE: To evaluate whether duration of HIV-1 infection influences the response to highly active antiretroviral therapy (HAART). DESIGN: Prospective study of individuals (Italian Seroconversion Study cohort) with well-estimated dates of HIV-1 seroconversion. METHODS: This analysis included 277 participants who began HAART (defined as three antiretroviral drugs used in combination). Cox regression models were used to evaluate the association between duration of infection (as categorical variable [7.5 years from seroconversion] or continuous variable) and an immunologic (rise in CD4 count >100 cells/mm3) and a virologic (decline in plasma HIV-RNA to unquantifiable levels) outcome. All analyses were stratified by center of recruitment and adjustment, when used, was for gender, age at inception of HAART, injection drug use, previous antiretroviral therapy, lag-time between positive and negative HIV test result, year of starting HAART, clinical stage, CD4 count, and HIV-RNA at time of HAART. RESULTS: HAART was initiated a median of 6.4 years after seroconversion. There was a median follow-up of 1.6 years after starting HAART to the calendar cut-off (November 1999). One-hundred-eighty-one (65.3%) patients experienced a decline in viral load to below quantifiable levels and 184 (66.4%) experienced a rise in CD4 >100 cells/mm3. In the Cox models, by 1-year increase in duration of infection, we estimated a lower crude hazard of achieving a CD4 count increase >100 cells (relative hazard [RH], 0.96; 95% confidence interval [CI], 0.92-1.01; p =.09), and a lower hazard of reaching an unquantifiable level of plasma HIV-RNA (RH, 0.97; 95%CI, 0.93-1.02; p =.20). After adjustment, these values became 0.99 (95%CI, 0.93-1.04; p =.62) and 0.98 (95%CI, 0.93-1.04; p =.48), respectively. When duration of HIV infection was considered as a categorical variable, the results were consistent with those already described. CONCLUSIONS: These results suggest that the duration of HIV infection does not seem to play an important independent role in determining the virologic and immunologic responses to HAART. PMID- 11391169 TI - Mortality in a cohort of HIV-infected adults started on a protease inhibitor containing therapy: standardization to the general population. AB - Death rates in the APROCO cohort of 1,157 HIV-1 infected adults starting for the first time a protease inhibitor-containing therapy were standardized to the 1996 French general population mortality rates stratified by age and gender. Median follow-up was 23 months and mortality rate was 2.2% person-years (95% confidence interval [CI] = 1.6-2.9). Overall mortality was 7.8 times higher than in the general population (95% CI = 5.7-10.4), 4.7 in men and 19.5 in women. Among the 144 patients considered complete responders, the death rate was 1.2% person-years (95% CI = 0.2-3.5) and mortality remained 5.1 times higher (95% CI = 1.0-14.9) than in the general population. Failure of treatment, long-term adverse effects, or less favorable socio-demographic status could explain these trends. PMID- 11391170 TI - Methadone maintenance as HIV risk reduction with street-recruited injecting drug users. AB - OBJECTIVE: To compare changes in HIV risk behaviors between street-recruited opiate injectors who entered and remained in methadone maintenance treatment and those who did not. METHODS: Three hundred sixteen participants were interviewed at baseline, received outreach interventions, and were interviewed again 6 months later. RESULTS: Significant (p <.001) reductions in HIV-related risk behaviors, including frequency of injecting, injecting with used (dirty) needles, and sharing injection paraphernalia, were demonstrated. Participants (31%) who entered and remained in methadone maintenance treatment for at least 90 days before follow-up showed a significantly greater reduction in heroin injections than those who did not. They did not show a greater reduction in using dirty needles or sharing other injection paraphernalia. CONCLUSIONS: These findings suggest that although methadone maintenance may reduce injection frequency, it does not reduce other HIV-related risk behaviors above and beyond what can be accomplished through outreach interventions. Treatment facilities and outreach intervention programs should collaborate to provide a comprehensive approach to reducing HIV risk behaviors among drug injectors both in and out of drug treatment. PMID- 11391171 TI - Human T-cell lymphotropic virus type 1 infection among pregnant women in northeastern Brazil. AB - An evaluation of human T-cell lymphotropic virus type 1 (HTLV-1) infection among 6754 pregnant women in Salvador, Bahia, Brazil using enzyme-linked immunosorbent assay, Western blot analysis, and polymerase chain reaction assay found a rate of infection of 0.84% (57 of 6754 women). Epidemiologic and obstetric data on the HTLV-1-positive pregnant women were analyzed and compared with data on a control group of HTLV-1-negative pregnant women. The mean age of the HTLV-1-positive women was 26.2 years. All were seronegative for HIV and syphilis, and only 2 reported a past history of sexually transmitted infection and more than 10 sexual partners. Of the HTLV-1-positive women, 88.5% were breast-fed, 4% were bottle fed, and 7.5% did not know. Six women had received blood transfusions, and only 1 reported intravenous drug use. Fifty-two HTLV-1-positive women could be followed: 45 had full-term deliveries, 5 had premature deliveries, and 2 had abortions. Our results indicate that (1) the frequency of HTLV-1 infection among pregnant women is relatively high in Salvador, Bahia, Brazil; (2) maternal infection was probably acquired more frequently through breast-feeding, but the sexual route was certainly the second most important means of transmission; (3) HTLV-1 positive women had a history of eczema-like infections in childhood more frequently than the control group; (4) HTLV-1 infection did not interfere in the course of pregnancy; and (5) no associated congenital infections were observed in the HTLV-1-positive women. PMID- 11391172 TI - Resistance to antiretroviral therapy among patients in Uganda. AB - OBJECTIVE: To characterize HIV-1 phenotypic resistance patterns and genotypic mutations among patients taking antiretroviral medications in Uganda. METHODS: We reviewed charts and retrieved archived plasma specimens from patients at an AIDS specialty center in Uganda where antiretroviral therapy has been used since 1996. Phenotypic and genotypic resistance testing was done on specimens associated with a viral load of 1000 copies/ml. RESULTS: Resistance testing of specimens was completed for 16 patients. Among 11 specimens collected before initiation of antiretroviral therapy, no phenotypic resistance or primary genotypic mutations were found. Among 8 patients taking lamivudine, phenotypic resistance was found for 9 (90%) of 10 specimens and was associated with an M184V mutation in all nine cases. Among 12 patients taking zidovudine, no phenotypic resistance and few primary mutations were found. For 6 patients who were receiving protease inhibitors, we observed no phenotypic resistance and only one primary genotypic mutation associated with resistance. CONCLUSIONS: The absence of apparent resistance among samples collected before antiretroviral therapy supports the notion that a similar approach to selection of antiretroviral therapy can generally be used against non-B subtypes. A genotypic marker of antiretroviral resistance to lamivudine in HIV-1 subtypes A, C, and D was similar to those in subtype B infections. These results suggest that the methods used for monitoring for the emergence of drug resistance in antiretroviral programs in Africa may be similar to those used in developed settings. PMID- 11391173 TI - High prevalence of genotypic and phenotypic HIV-1 drug-resistant strains among patients receiving antiretroviral therapy in Abidjan, Cote d'Ivoire. AB - To describe prevalence of antiretroviral (ARV) drug-resistant HIV-1 strains among patients with a history of earlier treatment with ARV drugs in Abidjan, Cote d'Ivoire, we determined mutations that confer HIV-1 ARV drug resistance by sequencing the viral reverse-transcriptase and protease genes derived from plasma viral RNA of 68 individuals consecutively enrolled in the Joint United Nations Program on AIDS Drug Access Initiative (UNAIDS-DAI) with a history of earlier ARV drug treatment in Abidjan between August 1998 and April 1999. Phenotypic ARV drug resistance was assessed using a recombinant virus assay. Primary mutations associated with ARV drug resistance to at least one of the reverse-transcriptase inhibitors or protease inhibitors were detected in 39 (57.4%) of the 68 patients. The prevalence of mutations associated with resistance to ARV drugs was: 29 (42.6%) to zidovudine, 10 (14.7%) to lamivudine, one (1.5%) to didanosine, one K103N mutation (associated with resistance to delavirdine, nevirapine, and efavirenz), one Y181C mutation (associated with resistance to delavirdine and nevirapine), two to both indinavir (M46I/L and V82A) and saquinavir (G48V and L90M), and one each to ritonavir (V82A) and nelfinavir (D30N). Phenotypic resistance to at least one nucleoside reverse transcriptase inhibitor (RTI) was seen in 25 (39.7%) patients, to nonnucleoside RTIs in 5 (8%) patients, and to protease inhibitors in 4 (6%) patients. The high prevalence we observed in this study may limit in future the effectiveness of ARV programs in the Cote d'Ivoire. PMID- 11391174 TI - Validation of genetic case-control studies in AIDS and application to the CX3CR1 polymorphism. AB - New polymorphisms have been recently identified in CX3CR1, a coreceptor for some HIV-1 strains, one of which was associated with a strong acceleration of HIV disease progression. This effect was observed both by a case-control study involving 63 nonprogressors (NP) from the asymptomatic long-term (ALT) cohort and Kaplan-Meier analysis of 426 French seroconverters (SEROCO cohort). These results prompted us to analyze these polymorphisms in 244 nonprogressors (NPs) and 80 rapid progressors (RPs) from the largest case-control cohort known to date, the GRIV cohort. Surprisingly, the genetic frequencies found were identical for both groups under all genetic models (p >.8). The discrepancy with the previous work stemmed only from the difference between GRIV NPs versus ALT NPs. We hypothesized this might be due to the limited number of NPs in ALT (n = 63) and in this line we reanalyzed the data previously collected on GRIV for over 100 different genetic polymorphisms: we effectively observed that the genetic frequencies of some polymorphisms could vary by as much as 10% (absolute percentage) when computing them on the first 50 NP subjects enrolled, on the first 100, or on all the NPs tested (240 study subjects). This observation emphasizes the need for caution in case-control studies involving small numbers of subjects: p values should be low or other control groups should be used.However, the association of the CX3CR1 polymorphism with progression seems quite significant in the Kaplan Meier analysis of the SEROCO cohort (426 individuals), and the difference observed with GRIV might be explained by a delayed effect of the polymorphism on disease. Further studies on other seroconverter cohorts are needed to confirm the reported association with disease progression. PMID- 11391175 TI - Curdlan sulfate induces the downmodulation of chemokine receptors leading to suppression of HIV infection. PMID- 11391176 TI - Lack of association between pregnancy and selected gastrointestinal adverse events among women prescribed nelfinavir. PMID- 11391177 TI - R211K and L214F do not invariably confer high level phenotypic resistance to thymidine analogs in zidovudine-naive patients with M184V. PMID- 11391178 TI - Syringe type preference among injection drug users. PMID- 11391179 TI - The reversal exchange technique of total calvarial reconstruction for sagittal synostosis. AB - The role of total calvarial reconstruction in the treatment of sagittal synostosis remains controversial, especially in patients younger than 1 year of age. The purpose of this study was to prospectively evaluate the efficacy of a single surgical technique for total calvarial reconstruction (the reversal exchange technique) in patients younger than 1 year of age who had a radiographically confirmed diagnosis of sagittal synostosis. Twenty-three consecutive patients underwent the reversal exchange technique of total calvarial reconstruction at a median age of 3 months (age range, 6 weeks to 10 months). Quantitative assessments were performed on the basis of preoperative and postoperative (minimum, 6 months) measurements of the cephalic index (cranial width/cranial length x 100) taken from three-dimensional computed tomography scans, which were obtained in 18 of 23 patients. Aesthetic assessments were performed on the basis of the grading of preoperative and postoperative photographs, obtained in 17 of 23 patients, by three independent raters who were blinded as to the surgical technique. The mean preoperative cephalic index was 65.0, and the mean postoperative index was 76.4, yielding a mean improvement of 11.4 (17.5 percent). By photographic evaluation, 12 of 17 patients (70.6 percent) were classified as having a normal head shape (grade 4) and five of 17 (29.4 percent) as having minor residual deformities (grade 3). No patients were identified as having significant residual deformities (grades 1 or 2). There were two intraoperative complications and one postoperative complication, none of which resulted in permanent morbidity. It was concluded that the reversal exchange technique of total calvarial reconstruction provided significant improvement in head shape on the basis of quantitative measurements (cephalic index) and independent evaluations of aesthetic improvement. PMID- 11391180 TI - Noninvoluting congenital hemangioma: a rare cutaneous vascular anomaly. AB - The authors studied a rare, congenital, cutaneous vascular anomaly that grows proportionately with the child and does not regress. A total of 53 patients were compiled from three vascular anomaly centers. These patients' lesions were analyzed for presentation, physical findings, radiologic and histopathologic characteristics, natural history, and outcome after resection. The lesions occurred slightly more often in male patients, always appeared alone, and were located (in order of frequency) in the head/neck region, extremities, and trunk. They were round-to-ovoid in shape, were plaque-like or bossed, occurred in variable shades of pink to purple, and had an average diameter of 5 cm. The overlying skin was frequently punctuated by coarse telangiectasia, often with central or peripheral pallor. The lesions were warm on palpation; fast-flow was further documented by Doppler ultrasonography. Magnetic resonance imaging and angiographic findings were similar to those of common hemangioma of infancy. All lesions were easily excised without recurrence.Histologic examination revealed lobular collections of small, thin-walled vessels with a large, often stellate, central vessel. Interlobular areas contained predominantly dilated, often dysplastic veins; arteries were also increased in number. Small arteries were observed "shunting" directly into lobular vessels or into abnormal extralobular veins. "Hobnailed" endothelial cells lined the small intralobular vessels. Mast cells were increased. Tests for glucose transporter-1, a recently reported reliable marker for common hemangioma of infancy, were negative in all 26 specimens examined. In conclusion, the authors think these clinicopathologic and radiologic features define a rare vascular lesion for which the term "noninvoluting congenital hemangioma" is proposed. These lesions of intrauterine onset may be a variant of common hemangioma of infancy or another hemangiomatous entity with persistent fast-flow. PMID- 11391181 TI - Traumatic optic neuropathy: a review of 61 patients. AB - The outcome of traumatic optic neuropathy was evaluated following penetrating and blunt injuries to assess the effect of treatment options, including high-dose steroids, surgical intervention, and observation alone. Factors that affected improvement in visual acuity were identified and quantified. Sixty-one consecutive, nonrandomized patients presenting with visual loss after facial trauma between 1984 and 1996 were assessed for outcome. Pretreatment and posttreatment visual acuities were compared using a standard ophthalmologic conversion from the values of no light perception, light perception, hand motion, finger counting, and 20/800 down to 20/15 to a logarithm of the minimum angle of resolution (log MAR). The percentage of patients showing visual improvement and the degree of improvement were calculated for each patient group and treatment method. Measurements of visual acuity are in log MAR units +/- standard error of the mean. Patients who sustained penetrating facial trauma (n = 21) had worse outcomes than patients with blunt trauma (n = 40). Improvement in visual acuity after treatment was seen in 19 percent of patients with penetrating trauma compared with 45 percent of patients with blunt trauma (p < 0.05). Furthermore, patients with penetrating trauma improved less than those with blunt trauma, with a mean improvement of 0.4 +/- 0.23 log MAR compared with 1.1 +/- 0.24 in blunt trauma patients (p = 0.03). The patients with blunt trauma underwent further study. There was no significant difference in improvement of visual acuity in patients treated with surgical versus nonsurgical methods; however, 83 percent of patients without orbital fractures had improvement compared with 38 percent of patients with orbital fractures (p < 0.05). The mean improvement in patients without orbital fractures was 1.8 +/- 0.65 log MAR compared with 0.95 +/- 0.26 in patients with orbital fractures (p = 0.1). Twenty-seven percent of patients who had no light perception on presentation experienced improvement in visual acuity after treatment compared with 100 percent of patients who had light perception on admission (p < 0.05). The mean improvement in patients who were initially without light perception was 0.85 +/- 0.29 log MAR compared with 1.77 +/- 0.35 in patients who had light perception (p < 0.05). There were no significant differences in improvement of visual acuity when analyzing the effect of patient age and timing of surgery. Patients who sustain penetrating trauma have a worse prognosis than those with blunt trauma. The presence of no light perception and an orbital fracture are poor prognostic factors in visual loss following blunt facial trauma. It seems that clinical judgment on indication and timing of surgery, and not absolute criteria, should be used in the management of traumatic optic neuropathy. PMID- 11391182 TI - Anatomic microstructure of the upper eyelid in the Oriental double eyelid. AB - The anatomic differences in the microstructure of the upper eyelid between the double eyelid and the nondouble eyelid are compared to determine the mechanism of double eyelid formation. Tissue from the upper eyelids of normal adult women was categorized into three groups: in one group, the double eyelid was formed primarily (at birth); in a second group, the double eyelid was formed gradually; and those in a third group had nondouble eyelids. A total of 56 eyelids were studied using electron microscopy and light microscopy. The results indicated that there is a significant difference between the three groups using scanning electron microscopy. In the upper eyelid of the double eyelid, bunched fibers of levator aponeurosis penetrate through orbicularis muscle to fuse with the skin in palpebral sulcus. This structure was not observed in the group with nondouble eyelids. However, when using light microscopy, this disparity was not observed. It was concluded that a fiber-linked structure between eyelid skin and levator aponeurosis is essential for the formation of the double eyelid. PMID- 11391183 TI - A new classification of palatal fracture and an algorithm to establish a treatment plan. AB - Palatal fractures have previously been classified according to the anatomic location of the fracture line, which is helpful for understanding the types of palatal fracture, but which is insufficient for helping the surgeon to decide which fracture to open and how to do so. The purpose of this study was to aid in the establishment of a precise treatment plan by determining the surgical approach and the types of stabilization that should be used for different types of palatal fracture. In a retrospective review of 136 consecutive Le Fort maxillary fractures over 6 years, 18 patients (13.2 percent) with palatal fractures were analyzed. The principle of open reduction and internal fixation was applied to all the patients. In six patients (33 percent), exploration and fixation was done in the palatal surface. Eight patients (44 percent) needed an extended period of immobilization (4 to 6 weeks). No major complications were observed during the follow-up period. An algorithm was devised to help establish a proper treatment plan, and palatal fractures were classified into four types: closed reduction, anterior treatment, anterior and palatal treatment, and combined. The key elements considered in deciding the treatment principle and the classification of a palatal fracture were the possibility of closed reduction, surgical exposure, site of rigid fixation, and stability of fractured segments after rigid fixation. The outcome of reconstruction and the postoperative course differed depending on the type of palatal fracture. This classification scheme provided an easy and simple way to establish a treatment plan and was helpful in learning the treatment principles of palatal fracture. PMID- 11391185 TI - Bilateral small radial forearm flaps for the reconstruction of buccal mucosa after surgical release of submucosal fibrosis: a new, reliable approach. AB - Oral submucous fibrosis is a collagen disorder affecting the submucosal layer and often severely limiting mouth opening. Previous surgical treatments have been disappointing. This article introduces a new surgical approach: reconstructing the bilateral buccal mucosa with two small radial forearm flaps. The surgical method includes the complete surgical release of fibrotic buccal mucosa and, if necessary, a bilateral coronoidectomy and temporalis muscle myotomy. From 1997 to 1999, 15 patients with moderate-to-severe trismus received reconstructive surgery, for a total of 30 small radial forearm flaps after surgical release. The flap size was between 1.5 x 5 and 2.5 x 7 cm. All donor sites were directly closed, and all flaps survived completely, except for one with partial necrosis. Six flaps required minor revisions because of size redundancy. Two patients developed buccal cancer in the area of reconstruction. At an average of 12 months' follow-up, the inter-incisal distance averaged 33 mm, an increase of 17 mm compared with the preoperative value. The donor-site morbidity was minimal, except in one heavy smoker who developed dry gangrene of his fingertips. The use of two small free forearm flaps for buccal mucosa reconstruction allows more radical release of fibrotic tissue. Coronoidectomy and temporal muscle myotomy further contribute to the effect of trismus release. The combined effects of this approach have consistently given good results. An aggressive approach toward surgical treatment of this precancerous lesion also facilitates the detection of cancer at an early stage. PMID- 11391186 TI - Necrotizing fasciitis of the head and neck: an analysis of 47 cases. AB - Necrotizing fasciitis is an overwhelming infection common to the perineum, abdominal wall, and extremities. It is a surgical emergency related to a high mortality rate that is more often seen in elderly and immunocompromised patients. Necrotizing fasciitis occurs uncommonly in the head and neck region. Over a 12 year period, 47 cases of necrotizing fasciitis of the head and neck region were collected at this hospital. The demographics, predisposing factors, clinical presentation and courses, management, complications, and outcomes were analyzed. The cases were divided into two groups: survivors and nonsurvivors. Statistical comparisons were made of the parameters age, gender, smoking or drinking habit, underlying medical problems, laboratory data, and treatments used. Forty-two patients (89.4 percent) had associated systemic disease; most of these patients had diabetes (72.3 percent). The clinical manifestations are nonspecific but are often typical for diagnosis. The necessity of computed tomographic scans is not conclusive in this study. Presentation of septic shock (p = 0.004) and association with underlying malignancy (p = 0.03) were the only statistically significant factors that led to a poor prognosis. The cornerstones of proper management include early diagnosis, aggressive surgical debridement, broad spectrum antibiotics, and intensive supportive care. PMID- 11391187 TI - Evaluation of the factors related to postmastectomy breast reconstruction. AB - A retrospective study was conducted in 75 consecutive patients requiring postmastectomy breast reconstruction over a period of 30 months. Each woman was offered one of the following four reconstructive options: free transverse rectus abdominis musculocutaneous flap (total number of reconstructions, n = 34); latissimus dorsi musculocutaneous flap (with or without expander and implant, n = 14); endoscopically assisted harvest of the latissimus dorsi muscle (with expander and implant, n = 13); and application of expander and implant only (n = 12). Of those patients originally selected for retrospective study, six did not meet the short-term prognostic criteria, and concerted attempts to contact two others proved unsuccessful. The remaining 67 patients were examined for the clinically assessed aesthetic appearance of the reconstructed breast(s), the subjective self-assessment of patient satisfaction, and the possible development of postoperative complications. Of these patients, six required bilateral surgery, which accounts for a final sample size of 73 individual breast reconstructions. The 67 individual patients were assessed after a minimum time of 6 months postreconstruction and became the sampling units for analysis. The free transverse rectus abdominis musculocutaneous flap procedure was the preferred method of breast reconstruction in 34 of 73 patients (47 percent), provided that it was generally agreed that the patient could endure a prolonged operation and that there was sufficient unscarred abdominal tissue available. Thereafter, postmastectomy radiotherapy at the chest wall became the primary criterion for assignment of a patient to a particular surgical procedure. Whenever radiotherapy resulted in poor-quality skin at the chest wall, endoscopically assisted transfer of latissimus dorsi muscle flap was considered to be the optimal treatment (13 of 73 patients, or 18 percent). Body mass index and smoking were secondary factors that were taken into account when this alternative technique was being considered.In the absence of radiotherapy, and provided that the chest wall was minimally scarred, patients who were reluctant to have reconstruction with autologous tissue were treated with expander and implant only (12 of 73, or 16 percent). This third procedure is a physically less arduous ordeal for the patient and was therefore the choice for all patients for whom a prolonged operation was not a realistic option. The fourth (and final) surgical procedure, latissimus dorsi musculocutaneous flap (with or without expander and implant), was selected for all patients with a better quality of skin over the chest wall, those whose abdomen was extensively scarred, and those who were on a general surgeon's operating list to undergo immediate breast reconstruction after mastectomy (14 of 73, or 19 percent). Equally good aesthetic results could be demonstrated with each of the four treatment options, provided that the reconstructive procedure selected was optimal for the individual patient and in accordance with the criteria described above. A variety of potential risk factors were considered for association with postoperative complications, including prescribed medication, obesity, smoking behavior, use of radiotherapy, and the recorded aggregated operative time. Of these, only body mass index (p < 0.001) and use of steroids (p = 0.016) were identified as having statistically significant effects on the incidence of adverse events.Finally, the general level of satisfaction expressed by the patient was highly correlated with a good appearance of the reconstructed breast, the physical comfort experienced while wearing a brassiere, and the general mobility of the unsupported reconstruction. PMID- 11391188 TI - Prospective evaluation of late cosmetic results following breast reconstruction: I. Implant reconstruction. AB - The long-term cosmetic outcome of breast implant reconstruction is unknown. The morbidity and cosmetic outcome of 360 patients who underwent immediate postmastectomy breast reconstruction with various types of implants have been analyzed prospectively over a 9-year period. Of these patients, 334 who completed their reconstruction were suitable for evaluation of their cosmetic outcome. The early complication rate (< 2 months) was 9.2 percent, with an explantation rate of 1.7 percent. The late complication rate (> 2 months) was 23 percent, with a pathological capsular contracture rate of 11 percent at 2 years and 15 percent at 5 years and an implant removal rate of 7 percent. The revisional surgery rate was 30.2 percent. The cosmetic results were assessed prospectively using an objective five-point global scale. Every patient was scored at each visit once surgery was completed. The overall cosmetic outcome deteriorated in a linear fashion, from an initial acceptable result of 86 percent 2 years after patients completed their reconstruction to only 54 percent at 5 years. This decline in cosmetic outcome was not associated with the type of implant used, the volume of the implant, the age of the patient, or the type of mastectomy incision employed. Radiotherapy was not a significant factor because only 28 patients were irradiated. Upon Cox model analysis, pathological capsular contracture was the only factor that contributed significantly to a poor cosmetic outcome in which p < 0.0001 (relative risk 6.3). Despite a high revisional surgery rate, deterioration still occurred, suggesting that other unaccounted for variables were responsible. On photographic retrospective review of the patients without capsular contracture who demonstrated deterioration in their cosmetic scores, it became clear that a possible reason for their poor results was late asymmetry produced by the failure of both breasts to undergo symmetrical ptosis with aging. PMID- 11391189 TI - Prospective evaluation of late cosmetic results following breast reconstruction: II. Tram flap reconstruction. AB - Although it is thought that transverse rectus abdominis muscle (TRAM) flap breast reconstruction produces excellent cosmetic results that are maintained over the long term, there is little objective evidence in the literature to support this. One hundred seventy-one consecutive patients who underwent TRAM flap reconstruction were prospectively analyzed over an 8-year period to assess their morbidity and late cosmetic outcome.The early patient complication rate (< 2 months) was 37.4 percent, the late hernia and fat necrosis rates (> 2 months) were 8.8 and 13.5 percent, respectively, and the contralateral symmetrization rate was 33.9 percent. The cosmetic results were evaluated prospectively using an objective five-point global scale. Each patient was scored at each visit once surgery was completed. Follow-up continued until a flap was lost, a patient died, or the point of last patient contact was reached. Six patients died during the study. The actuarial percentage cosmetic outcome remained stable during the study period, with an acceptable result in 96.4 percent of patients at 2 years and in 94.2 percent of patients at 5 years. Only five patients in this series obtained poor cosmetic outcomes, with three due to substantial flap necrosis and two because of poor flap design. Two free TRAM flaps were also lost. Log-rank analysis revealed that neither patient age nor timing of surgery significantly affected the cosmetic outcome. Single pedicle and supercharged (single pedicle) TRAM flaps produced slightly better results than bipedicle and free TRAM flaps. In this prospective longitudinal study, TRAM flap reconstructions were shown to produce aesthetically pleasing results. Moreover, with long-term follow-up, it was demonstrated that these reconstructions maintained their stability. PMID- 11391190 TI - Leprosy affects the tibial nerves diffusely from the middle of the thigh to the sole of the foot, including skip lesions. AB - This study investigated where leprosy affects the posterior tibial nerve and whether neurolysis is beneficial. Nine patients with bilateral posterior tibial leprous neuropathy with no sensorimotor recovery were studied. Preoperative sensory-muscle and nerve conduction velocity testing revealed the tarsal tunnel to be the site of a severe lesion in all cases. During surgery, the most proximal site of the nerve lesion was detected by electrically stimulating the spinal roots from the second lumbar nerve to the fourth sacral nerve, evoking efferent mixed nerve compound action potentials that were recorded from the exposed tibial nerve. In all patients, the nerve compound action potentials became normal only proximal to the sciatic nerve bifurcation. Epineuriotomy within these seemingly unaffected segments revealed fibrosis of the interfascicular epineurium. Interfascicular neurolysis was performed on all affected segments. A 2-year follow-up showed an increase in girth of the proximal calf musculature in six of eight patients (the ninth patient had no recordable nerve conduction velocity). It was concluded that (1) leprosy affects the tibial nerves in a scattered way from the sciatic nerve main trunk distally to the exit of the tarsal tunnel; and (2) interfascicular, microsurgical neurolysis is beneficial provided that it is performed on all affected nerve segments. PMID- 11391191 TI - The adductor flap: a new method for transferring posterior and medial thigh skin. AB - Skin flaps from the medial aspect of the thigh have traditionally been based on the gracilis musculocutaneous unit. This article presents anatomic studies and clinical experience with a new flap from the medial and posterior aspects of the thigh based on the proximal musculocutaneous perforator of the adductor magnus muscle and its venae comitantes. This cutaneous artery represents the termination of the first medial branch of the profunda femoris artery and is consistently large enough in caliber to support much larger skin flaps than the gracilis musculocutaneous flap. In all 20 cadaver dissections, the proximal cutaneous perforator of the adductor magnus muscle was present and measured between 0.8 and 1.1 mm in diameter, making it one of the largest skin perforators in the entire body. Based on this anatomic observation, skin flaps as large as 30 x 23 cm from the medial and posterior aspects of the thigh were successfully transferred. Adductor flaps were used in 25 patients. On one patient the flap was lost, in one the flap demonstrated partial survival, and in 23 patients the flaps survived completely. The flap was designed as a pedicle island flap in 14 patients and as a free flap in 11. When isolating the vascular pedicle for free tissue transfer, the cutaneous artery is dissected from the surrounding adductor magnus muscle and no muscle is included in the flap. Using this maneuver, a pedicle length of approximately 8 cm is isolated. In addition to ample length, the artery has a diameter of approximately 2 mm at its origin from the profunda femoris artery. The adductor flap provides an alternative method for flap design in the posteromedial thigh. Because of the large pedicle and the vast cutaneous territory that it reliably supplies, the authors believe that the adductor flap is the most versatile and dependable method for transferring flaps from the posteromedial thigh region. PMID- 11391192 TI - Management of soft-tissue problems in leg trauma in conjunction with application of the Ilizarov fixator assembly. AB - Forty-five patients presenting with high-energy open grade III tibial diaphyseal fractures were treated with the Ilizarov technique. Of these patients, 28 required plastic surgical intervention for achieving wound closure. Most of the injuries were complicated by initial neglect and inadequate primary soft-tissue coverage resulting in osteitis, sequestration, and segmental diaphyseal tibial defects, often in combination with skin-envelope deficits of various types in and around the fracture perimeter. The unique soft-tissue problems encountered while using the Ilizarov fixator have not been focused on in previous reports on the management of segmental bone defects. Four basic local flap procedures: the transposition flap, rotation flap, adipofascial turnover flap, and Z-plasty are useful and versatile for managing most types and grades of soft-tissue defects associated with a segmental bone loss with the Ilizarov technique. PMID- 11391193 TI - Laser Doppler flowmeter monitoring of free-tissue transfers: blood flow in normal and complicated cases. AB - In this article, laser Doppler flowmeter (LDF) monitoring of blood flow in 94 free flaps is summarized. Seventy-six patients had uneventful postoperative courses, and 18 patients developed postoperative complications, with a salvage rate of 88 percent. Except for one case, the flowmeter identified developing complications before clear clinical indices appeared, and in two cases it was the only indication of vascular compromise of the flap. On the basis of the data obtained, the ranges of absolute flow values in different types of uncomplicated flaps are reported, along with their temporal pattern of flow. Decrease inflow pattern may be an early indicator of a developing perfusion disturbance. On the basis of LDF readings, the following classifications of free-flap blood flow are suggested. (1) If the flow is within or slightly above the established range, then normal diligence in observation is justified. If the flow is well above the normal range, artifacts that could falsely elevate readings should be investigated. (2) If the flow is somewhat below the established range, then a modest increase in observation is warranted (alert level 1). (3) If the relative flow falls to 50 percent of the initial flow of that flap and remains at that level for 30 minutes or longer, then more aggressive flap observation is indicated (alert level 2). (4) If the flow is below 0.4 LDF units for 30 minutes, then aggressive clinical observation should be performed (alert level 3, or "red alert") and exploration should be strongly considered. Falsely elevated measurements can be caused by vibration, motion of the probe or tissue, or location of the probe over a macroscopic blood vessel. False low readings are quite rare but can result from partial probe detachment from the flap or coagulum accumulating on the probe. Once artifacts are ruled out, LDF readings have a high level of credibility and, in the authors' experience, significantly improve salvage rates. PMID- 11391194 TI - The long head of the triceps brachii as a free functioning muscle transfer. AB - This anatomic study investigates the possibility of using the long head of the triceps brachii muscle as a free functioning muscle transfer for the upper limb. It has been reported that the long head is not difficult to harvest and that its loss does not create significant donor-site morbidity. The muscle was studied in 23 fresh frozen upper limbs. The long head in all 23 specimens had a constant and proximal vascular pedicle from the profunda brachii artery and vein. The mean pedicle was long (4 cm) and had large-caliber vessels (diameter, 3-mm artery and 4-mm vein). Angiograms were carried out in five specimens and dye perfusion studies in six specimens. A single branch from the radial nerve of at least 7 cm in length innervated the muscle. Muscle architecture was studied in 12 specimens and revealed that the long head of the triceps is better suited for forearm reconstruction than either the gracilis or the latissimus dorsi muscles. The mean physiologic cross-sectional area (8.36 cm(2)) and fiber length (10.8 cm on the superficial surface and 8.2 cm on the deep surface) of the long head match more closely those of the flexor digitorum profundus and the extensor digitorum communis, the muscles most commonly replaced. PMID- 11391195 TI - Free medial plantar perforator flaps for the resurfacing of finger and foot defects. AB - In this article, three cases in which free medial plantar perforator flaps were successfully transferred for coverage of soft-tissue defects in the fingers and foot are described. This perforator flap has no fascial component and is nourished only by perforators of the medial plantar vessel and a cutaneous vein or with a small segment of the medial plantar vessel. The advantages of this flap are minimal donor-site morbidity, minimal damage to both the posterior tibial and medial plantar systems, no need for deep dissection, the ability to thin the flap by primary removal of excess fatty tissue, the use of a large cutaneous vein as a venous drainage system, a good color and texture match for finger pulp repair, short time for flap elevation, possible application as a flow-through flap, and a concealed donor scar. PMID- 11391196 TI - Free tensor fasciae latae perforator flap for the reconstruction of defects in the extremities. AB - In the three cases presented in this study, free tensor fasciae latae perforator flaps were used successfully for the coverage of defects in the extremities. This flap has no muscle component and is nourished by muscle perforators of the transverse branch of the lateral circumflex femoral system. The area of skin that can by nourished by these perforators is larger than 15 x 12 cm. The advantages of this flap include minimal donor-site morbidity, the preservation of motor function of the tensor fasciae latae muscle and fascia lata, the ability to thin the flap by removing excess fatty tissue, and a donor scar that can be concealed. In cases that involve transection of the perforator above the deep fascia, the operation can be completed in a very short period of time. This flap is especially suitable as a free flap for young women and children who have scars in the proximal region of the lateral thigh or groin region that were caused by split-thickness skin grafting or full-thickness skin grafting during previous operations. PMID- 11391197 TI - Free anterolateral thigh flap for extremity reconstruction: clinical experience and functional assessment of donor site. AB - From August of 1995 through July of 1998, 38 free anterolateral thigh flaps were transferred to reconstruct soft-tissue defects. The overall success rate was 97 percent. Among 38 anterolateral thigh flaps, four were elevated as cutaneous flaps based on the septocutaneous perforators. The other 34 were harvested as myocutaneous flaps including a cuff of vastus lateralis muscle (15 to 40 cm3), either because of bulk requirements (33 cases) or because of the absence of a septocutaneous perforator (one case). However, vastus lateralis muscle is the largest compartment of the quadriceps, which is the prime extensor of the knee. Losing a portion of the vastus lateralis muscle may affect knee stability. Objective functional assessments of the donor sites were performed at least 6 months postoperatively in 20 patients who had a cuff of vastus lateralis muscle incorporated as part of the myocutaneous flap; assessments were made using a kinetic communicator machine. The isometric power test of the ratios of quadriceps muscle at 30 and 60 degrees of flexion between donor and normal thighs revealed no significant difference (p > 0.05). The isokinetic peak torque ratio of the quadriceps and hamstring muscles, including concentric and eccentric contraction tests, showed no significant difference (p > 0.05), except the concentric contraction test of the quadriceps muscle, which revealed mild weakness of the donor thigh (p < 0.05). In summary, the functional impairment of the donor thighs was minimal after free anterolateral thigh myocutaneous flap transfer. PMID- 11391198 TI - Sparing a testis during vaginoplasty in male-to-female transsexuals: does it benefit our patients? AB - Testosterone and its metabolite dihydrotestosterone are the libido hormones for the male, vital to sex drive and sexual function. Fear of loss of libido and orgasm is the main reason to retain at least one testis in male-to-female transsexuals during vaginoplasty. We report on four South American transsexual patients in whom we resected a remaining testis to illustrate the superfluity of retaining it. Because there are multiple reasons for castration, we advise that bilateral orchidectomy be performed in the course of sex reassignment surgery for male-to-female transsexuals. PMID- 11391199 TI - Immunolocalization of fibroblast growth factor receptors 1 and 2 in mouse palate development. AB - Recent evidence has implicated mutations of fibroblast growth factor receptors (FGF-R) in the pathogenesis of craniosynostotic syndromes. Cleft palate can be a component of such syndromes. The expression of FGF-R1 and FGF-R2 has been delineated in normally developing cranium, where they seem to regulate cellular differentiation and proliferation, respectively. The specific role of fibroblast growth factor signaling in mammalian palate development is unclear. The authors investigated the patterns of expression of FGF-R1 and FGF-R2 throughout mouse palatal development in the embryo. Time-dated CD-1 mouse heads (n = 135) were harvested at embryonic ages 12.5, 13.5, 14.5, 15.5, and 16.5 days (term gestation = 19.5 days), fixed in paraformaldehyde, embedded in paraffin, and sectioned. In addition, paired palatal shelves (n = 30) were isolated by means of microdissection from embryonic day--13.5 embryos, grown on Millipore filters in serum-free medium in vitro for 24, 48, 72, or 96 hours and processed for histological analysis. Immunohistochemical analysis for FGF-R1 and FGF-R2 was performed on the in vivo and in vitro specimens. FGF-R1 and FGF-R2 were found to be specifically expressed in the epithelium of the developing palatal shelves from the time of their outgrowth from the maxillary processes through completion of fusion in vivo and in vitro. Expression of both receptors was particularly strong during the phases of medial epithelial-medial epithelial contact between the individual shelves, through the formation of the medial epithelial seam, to the ultimate dissolution of the seam. Such a pattern of expression seems to implicate fibroblast growth factor signaling in the regulation of the critical phase of fusion of the bilateral shelves. The expression of both FGF-R1 and FGF R2 in the lateral palatal mesenchyme, where such secondary structures as tooth primordia and bone begin to appear, also suggests a role for fibroblast growth factor signaling in the induction of ongoing differentiation and maturation of the palate after fusion. These data suggest that fibroblast growth factor signaling may play a role in the epithelial-mesenchymal interactions that dictate fusion and maturation of the developing palate. Furthermore, the data are consistent with the correlation of cleft palate formation with aberrant fibroblast growth factor signaling. PMID- 11391201 TI - Ontogeny of expression of transforming growth factor-beta 1 (TGF-beta 1), TGF beta 3, and TGF-beta receptors I and II in fetal rat fibroblasts and skin. AB - Fetal cutaneous wounds that occur in early gestation heal without scar formation. Although much work has been done to characterize the role of transforming growth factor-beta (TGF-beta) isoforms in the adult wound repair process, their function in fetal scarless wound repair is not well understood. The authors hypothesized that the pattern of expression for TGF-beta isoforms and their receptors may influence the phenotypic transition from scarless to scar-forming repair observed during fetal gestation. Using time-dated fetal Sprague-Dawley rat fibroblasts and unwounded skin at gestational ages 14, 16, 18, and 21 days postcoitum of the scarless (< or =16 days) and scar-forming (>16 days) periods of gestation (term = 21.5 days), the authors analyzed the endogenous messenger RNA (mRNA) levels of TGF-beta 1 and TGF-beta 3 and their signaling receptors TGF-beta-RI and TGF-beta RII. Northern blot analyses in both fibroblasts and unwounded skin revealed that levels of TGF-beta 1 were not differentially expressed, whereas more TGF-beta 3 mRNA transcript was found in early than in late gestation. Fibroblast expression of TGF-beta-RI showed no substantial differences, whereas expression of TGF-beta RII increased during gestation. In contrast, expression of both TGF-beta-RI and TGF-beta-RII in unwounded skin showed decreasing levels as a function of gestational age. The differential levels of TGF-beta 1 and TGF-beta 3 suggest that the ratio of these cytokines may provide a predominantly antiscarring or profibrotic signal upon wounding during the scar-free or scar-forming periods of gestation, respectively. Furthermore, lower amounts of the ligand-binding TGF beta-RII seen in early gestation fibroblasts suggest a decreased ability to perceive ligand during the period of scarless repair. PMID- 11391203 TI - Massive spontaneous hemothorax following bilateral reduction mammaplasty. PMID- 11391204 TI - Replantation of avulsed penoscrotal skin. PMID- 11391205 TI - Cultured autologous outer root sheath cells: a new therapeutic alternative for chronic decubitus ulcers. PMID- 11391206 TI - Reconstruction of a total Achilles tendon and soft-tissue defect using an Achilles allograft combined with a rectus muscle free flap. PMID- 11391207 TI - Distraction osteogenesis of the craniofacial skeleton. AB - Distraction osteogenesis is becoming the treatment of choice for the surgical correction of hypoplasias of the craniofacial skeleton. Its principle is based on the studies of Ilizarov, who showed that osteogenesis can be induced if bone is expanded (distracted) along its long axis at the rate of 1 mm per day. This process induces new bone formation along the vector of pull without requiring the use of a bone graft. The technique also provides the added benefit of expanding the overlying soft tissues, which are frequently deficient in these patients. This article reviews the authors' 11-year clinical and research experience with mandibular distraction osteogenesis. It highlights the indications and contraindications of the technique and emphasizes the critical role that basic science research has played in its evolution. PMID- 11391208 TI - Beware of fire! PMID- 11391209 TI - Secondary carpal tunnel surgery. AB - A small but significant group of patients with carpal tunnel syndrome "fail" primary carpal tunnel release and require secondary surgery. The persistence or recurrence of previous symptoms or the development of new symptoms is often indicative of the nature of the patient's problem. Postoperative complications may be classified into the general areas of neurological, vascular, tendon, and wrist complaints. A thorough clinical evaluation, including a complete neurological examination of the hand and upper extremity, provides an accurate assessment of the status of the median nerve. Important surgical techniques that may be used during secondary carpal tunnel surgery include internal neurolysis, neuroma-in-continuity assessment, neuroma management, nerve grafting, and tissue interposition flaps. PMID- 11391210 TI - The market of plastic surgery: cosmetic surgery for sale--at what price? PMID- 11391211 TI - Complications of abdominoplasty in 86 patients. AB - A total of 101 consecutive abdominoplasty patients were reviewed retrospectively. Of these, 14 male (mean age at time of operation, 34.3 years; range, 23 to 53 years) and 72 female (mean age at time of operation, 38.9 years; range, 19 to 64 years) patients had adequate documentation for inclusion in this study. Complications were recorded as either wound complications (wound infection, partial wound dehiscence, seroma, hematoma, and skin edge necrosis) or complications after surgery (deep vein thrombosis, pulmonary emboli, ileus, sensibility disorder of the skin of the thighs, and death). The complications were subsequently correlated for sex, race, the patient's age at surgery, body mass index before surgery, and the seniority of the surgeon. Nine male patients (64.3 percent) and 11 female patients (15.3 percent) had wound complications. Almost 10 percent of our patients sustained an injury to the lateral cutaneous nerve of the thigh. Male patients should be informed about their possible higher risk of complications, and special attention must be given by the surgeon to the prevention of such complications.Moreover, specific attention must be given to the preservation of the lateral cutaneous nerves of the thigh in both male and female patients undergoing abdominoplasties. PMID- 11391212 TI - Rhinoplasty: large nostril/small tip disproportion. AB - Surgeons must recognize large nostril/small tip disproportion as a distinct challenge in rhinoplasty surgery. The critical first step is to correctly analyze the intrinsic and extrinsic factors that contribute to the deformity. The nostril axis is drawn between the nostril apices and extended in both directions. It is then subdivided into a nostril and intrinsic tip component. The ratio of nostril to tip should be 55:45; a ratio of 60:40 is acceptable. The surgical solution requires both an increase in intrinsic tip projection by lengthening the infralobular segment and a nostril reduction. The anatomical deformity consists of three components: (1) the alar cartilages are highly divergent, (2) the infralobular segment is quite short, and (3) the domal segment is flat and ill defined. The operative technique advocated by the author combines a three-stitch tip procedure, including an interdomal suture over a straight strut, plus nostril sill/alar wedge resections. PMID- 11391214 TI - Laser blepharoplasty for making double eyelids in Asians. AB - The double-eyelid operation is a cosmetic procedure performed primarily on Asians who have no supratarsal folds. The goal in Oriental blepharoplasty is to ensure a stable double-eyelid fold with predictable and long-lasting results. Candidates for this procedure desire attractive eyelids with a natural-looking fold. Today, the CO2 laser is used as a surgical tool by plastic surgeons, and its use in cosmetic surgery has recently been expanded to blepharoplasties. We used the high power CO2 laser (UltraPulse; Coherent, Santa Clara, Calif.) in the double-eyelid operation. Between September of 1995 and September of 1999, a total of 241 patients underwent laser double-eyelid operations at Korea University Medical Center and Dr. Choi's Aesthetic Clinic, with an average follow-up of 18 months. By using the CO2 laser, we could create a stable double-eyelid fold with long lasting results. The rate of fold release was 2 percent, which was less than the rate using the conventional scalpel incision method. The CO2 laser contributed to a reduction in pain or discomfort after the operation; therefore, patient comfort and early recovery could be enhanced after double-eyelid surgery. In the healing of the incision line, it showed mild redness for 3 weeks, but long-term follow-up showed the scar produced by the laser appeared to be equal in quality compared with the conventional method. Laser blepharoplasty is a useful and effective method to create a stable and attractive double eyelid for Asian patients. PMID- 11391215 TI - Electrosurgical ablation: a new mode of cutaneous resurfacing. PMID- 11391216 TI - The greatest myths in breast augmentation. PMID- 11391219 TI - Role of donor kidney biopsies in renal transplantation. PMID- 11391220 TI - A predictive test for the efficacy of intravenous immunoglobulin in the inhibition of alloantibodies. AB - Preformed recipient HLA-specific antibodies can cause hyperacute rejection of a transplanted kidney if they are directed against mismatched donor HLA antigens. To avoid hyperacute rejection it is essential that recipient antibodies be identified during patient workup for transplantation and HLA antigens to which a patient is sensitized then be avoided when selecting a kidney donor for them. Absence of donor-specific reactivity is then confirmed by a pretransplantation crossmatch test. PMID- 11391221 TI - Induction of transplantation tolerance with a short course of tacrolimus (FK506): I. Rapid and stable tolerance to two-haplotype fully mhc-mismatched kidney allografts in miniature swine. AB - BACKGROUND: Inbred miniature swine provide a large animal model in which the effects of selective major histocompatibility complex (MHC) matching can be reproducibly studied. We have previously demonstrated that although a 12-day course of cyclosporine uniformly induces tolerance to class I-mismatched renal allografts, it does not induce tolerance across full MHC barriers. In this study, we assessed whether and at what dose tacrolimus might permit allografts to induce tolerance across different MHC barriers. METHODS: Recipients of MHC disparate renal allografts were treated with a 12-day course of tacrolimus by continuous intravenous infusion. Groups were divided as follows: (1) class I-mismatched kidneys with 0.3 mg/kg/day tacrolimus (n=3); (2) fully MHC-mismatched kidneys with 0.3 mg/kg/day tacrolimus (n=2); and (3) fully MHC-mismatched kidneys with 0.12-0.16 mg/kg/day tacrolimus (n=4). RESULTS: In groups 1 and 2, recipients with tacrolimus levels of 45-80 ng/ml accepted renal allografts long-term with stable renal function. Donor-specific hyporesponsiveness was demonstrated by cell mediated lymphocytotoxicity and mixed lymphocyte response, and subsequent donor matched grafts were also accepted, without further immunosuppression (n=4), confirming systemic tolerance. In group 3, recipients that achieved tacrolimus levels of 35 ng/ml (n=2) accepted their grafts without chronic changes, whereas recipients with levels of 20-26 ng/ml (n=2) developed chronic allograft glomerulopathy, suggesting 35 ng/ml as the threshold blood level for tolerance induction. In vitro assays demonstrated that peripheral blood lymphocytes from tolerant animals produced inhibitory cytokines, suggesting the involvement of regulatory mechanisms. CONCLUSIONS: To our knowledge, this study represents the first demonstration of the induction of transplant tolerance across a two haplotype full MHC barrier with a short course of immunosuppression in a large animal model. These studies may also have clinical applicability, because the time course required to induce tolerance was sufficiently short that the high drug levels required might be expected to be tolerated clinically with only transient toxicity. PMID- 11391222 TI - Heart preservation with celsior solution improved by the addition of nitroglycerine. AB - BACKGROUND: Preservation of rat hearts was extended to 16 hr when nitroglycerine (NTG) was added to colloid-free University of Wisconsin solution (MUW). This study examined the effectiveness of Celsior solution (CEL) and whether adding NTG to CEL would improve and extend cardiac preservation. METHODS: Two studies were conducted: (a) 9-hr preservation with either CEL or MUW, (b) 16-hr preservation with CEL, CEL+NTG, or MUW+NTG. Rat heart isografts were flushed and stored at 0 degrees C before heterotopic transplantation with an indwelling externalized intraventricular balloon-tipped catheter. One and 7 days after transplantation, quantitative functional studies were performed. RESULTS: After 9-hr preservation, all (6/6) grafts preserved with MUW beat for 7 days, whereas only 1/6 hearts preserved with CEL continued to beat. After 16-hr preservation, 6/10 CEL+NTG hearts beat for 7 days compared with 7/8 MUW+NTG hearts; none of the hearts preserved with CEL survived. Function was similar in CEL+NTG and MUW+NTG groups on day 1 (left ventricular developed pressure (LVDP): CEL+NTG=64+/-16, MUW+NTG=104+/-16 mmHg; maximum dP/dt: CEL+ NTG=2024+/-551, MUW+NTG=3582+/-513 mmHg/sec) and day 7: (LVDP: CEL+NTG=126+/-25, MUW+NTG=177+/-24 mmHg; maximum dP/dt: CEL+NTG=3835+/-848, MUW+ NTG=5639+/-670 mmHg/sec). Function in both groups improved significantly (P<0.05) on day 7 compared with day 1. CONCLUSIONS: Celsior was not as effective as MUW for rat heart preservation. The addition of NTG to both CEL and MUW provided similar effective preservation for 16 hr. NTG should be added routinely to both solutions. PMID- 11391223 TI - Lewis rat pancreas, but not cardiac xenografts, are resistant to anti-gal antibody mediated hyperacute rejection. AB - BACKGROUND: The objective of this study was to evaluate the role of anti-Gal Abs and non-anti-Gal Abs in hyperacute rejection (HAR) of concordant pancreas xenografts compared with heart xenografts. In addition, we tested whether rejection of Lewis rat pancreas grafts was T-cell dependent and could be prevented by anti-T-cell treatment. METHODS: To determine the role of anti-Gal Abs in the induction of HAR, Lewis rat pancreas and heart xenografts were transplanted into alpha1,3Galactosyltransferase knockout (GT-Ko) mice treated with normal human serum (NHS) or hyperimmune serum, or into presensitized GT-Ko mice. To investigate whether rejection of pancreas xenograft was mediated by a T cell dependent response, Lewis rat pancreas grafts were transplanted into streptozotocin (STZ)-induced diabetic GT-Ko mice treated with FK506, anti-CD4 mAbs (GK1.5), and thymectomy. Antidonor-specific IgM and IgG and anti-Gal Abs were analyzed by flow cytometry. Rejected and long-term surviving pancreas xenografts were assessed by functional (blood glucose) and histopathological examination. RESULTS: HAR of Lewis rat pancreas xenografts could not be induced by NHS (0.4 ml), whereas NHS (0.2 ml) resulted in HAR of Lewis heart xenografts. Infusion of Lewis rat-specific hyperimmune serum (0.2 ml) resulted in HAR of Lewis rat pancreas xenografts. In addition, second Lewis rat pancreas grafts were hyperacutely rejected by presensitized GT-Ko mice. Immunohistochemical staining showed a low expression of Galalpha1,3Gal antigen in the endocrine tissue compared with that in the cardiac grafts. The levels of anti-Gal Abs in pancreas xenograft transplantation did not increase in GT-Ko mice after pancreas xenograft transplantation that was significantly increased after heart transplantation. FK506 treatment induced long-term survival of Lewis pancreas xenografts (mean survival time (MST) >90 days). Anti-CD4 treatment delayed rejection of Lewis rat pancreas xenografts with MST of 34.3 days, whereas anti-CD4, in combination with thymectomy, synergistically prolonged survival of pancreas xenograft (MST=70.4 days). CONCLUSION: Pancreas xenograft is resistant to anti-Gal Abs-induced HAR but is susceptible to anti-donor specific Abs. Rejection of Lewis pancreas xenograft in STZ-induced, diabetic, GT-Ko mice is T-cell dependent. PMID- 11391224 TI - Peritubular capillary basement membrane reduplication in allografts and native kidney disease: a clinicopathologic study of 278 consecutive renal specimens. AB - BACKGROUND: An association has been found between transplant glomerulopathy (TG) and reduplication of peritubular capillary basement membranes (PTCR). Although such an association is of practical and theoretical importance, only one prospective study has tried to confirm it. METHODS: We examined 278 consecutive renal specimens (from 135 transplants and 143 native kidneys) for ultrastructural evidence of PTCR. In addition to renal allografts with TG, we also examined grafts with acute rejection, recurrent glomerulonephritis, chronic allograft nephropathy and stable grafts ("protocol biopsies"). Native kidney specimens included a wide range of glomerulopathies as well as cases of thrombotic microangiopathy, malignant hypertension, acute interstitial nephritis, and acute tubular necrosis. RESULTS: We found PTCR in 14 of 15 cases of TG, in 7 transplant biopsy specimens without TG, and in 13 of 143 native kidney biopsy specimens. These 13 included cases of malignant hypertension, thrombotic microangiopathy, lupus nephritis, Henoch-Schonlein nephritis, crescentic glomerulonephritis, and cocaine-related acute renal failure. Mild PTCR in allografts without TG did not predict renal failure or significant proteinuria after follow-up periods of between 3 months and 1 year. CONCLUSIONS: We conclude that in transplants, there is a strong association between well-developed PTCR and TG, while the significance of mild PTCR and its predictive value in the absence of TG is unclear. PTCR also occurs in certain native kidney diseases, though the association is not as strong as that for TG. We suggest that repeated endothelial injury, including immunologic injury, may be the cause of this lesion both in allografts and native kidneys. PMID- 11391225 TI - Value of cardiac troponin I and T for selection of heart donors and as predictors of early graft failure. AB - BACKGROUND: Cardiac troponin I and T (cTnI and cTnT) are sensitive and specific markers of myocardial damage. We evaluated them for the selection of heart donors and as predictors of early graft failure after heart transplantation. METHODS: cTnI, cTnT, myoglobin, and creatine kinase (CK) levels and its isoenzyme MB (CKMB) activity and mass were measured in serum samples immediately before opening the pericardium from 126 consecutive brain-dead multi-organ donors over 10 years of age inspected by our harvesting team. Donors with serum creatinine >2.0 mg/dL (n=6) were excluded from the analysis. Donors for high-urgency status recipients (n=2) were also excluded. The remaining donors were retrospectively divided into three groups: group I (n=68), grafts with good function; group II (n=11), grafts with impaired function; and group III (n=39), grafts not accepted for transplantation. RESULTS: No differences in donor and recipient characteristics were found among the groups. The mean values of cTnI (0.36+/-0.88 microg/L, 4.45+/-3.28 microg/L, and 3.02+/-7.88 micog/L, respectively) and cTnT (0.016+/-0.029 microg/L, 0.134+/-0.114 microg/L, and 0.123+/-0.245 microg/L, respectively) were lower in group I when compared with groups II or III (cTnI: P<0.0001, P=0.018; cTnT: P<0.0001, P=0.012). The cTnI value was higher in group II compared with group III (P=0.023). The cTnT values were similar in groups II and III. A cTnI value >1.6 microg/L as a predictor of early graft failure had a specificity of 94%, and a cTnT value of >0.1 microg/L had a specificity of 99%. The odds ratio for the development of acute graft failure after heart transplantation was 42.7 for donors with cTnI >1.6 microg/L and 56.9 for donors with cTnT >0.1 microg/L. No differences of myoglobin, CKMB activity, or CKMB/CK ratio were found among the groups. CONCLUSIONS: Significantly higher cTnI and cTnT values were found in peripheral blood at the time of explantation in donors of hearts with subsequently impaired graft function and in not accepted donors. cTnI and cTnT are useful as additional parameters for heart donor selection. PMID- 11391226 TI - RAD in de novo renal transplantation: comparison of three doses on the incidence and severity of acute rejection. AB - BACKGROUND: The effects of three doses of RAD (40-O-[2-hydroxyethyl]-rapamycin), a novel macrolide with potent immunosuppressive and antiproliferative properties, on the incidence and severity of acute rejection episodes as well as its tolerability were evaluated in a dose-ranging study in de novo renal transplant recipients. METHODS: In this double-blind, parallel group, multicenter study, recipients were randomized to receive 1 mg, 2 mg, or 4 mg/day of RAD in combination with Neoral (cyclosporine, USP MODIFIED) and corticosteroids. The incidence and severity of biopsy-proven acute rejection episodes, graft survival, patient survival, infection rates, laboratory measurements, and adverse events were compared across groups after 6 months of therapy. RESULTS: Among the 103 recipients, patients receiving 1, 2, or 4 mg/day experienced a 32.4%, 14.7%, or 25.7% incidence of biopsy-proven acute rejection episodes within the first 6 months posttransplantation, respectively. Even though the study was not powered to demonstrate efficacy, the incidence of moderate and severe acute rejection episodes was found to be significantly lower among patients in the 2 mg and 4 mg/day groups than in the 1 mg/day group (P=0.002 and P=0.006, respectively). Overall graft and patient survival rates were excellent. RAD was generally well tolerated. Although blood lipid levels increased in all groups, changes were manageable with lipid-lowering agents and did not warrant discontinuation of study medication. The incidence of viral and fungal infections was low; however, it was higher among recipients treated with 4 mg/day. CONCLUSIONS: In combination with Neoral and corticosteroids, RAD doses of 2 mg and 4 mg/day resulted in lower rates of acute rejection episodes and efficacy failure than the 1 mg/day dose and were significantly more effective in reducing the severity of rejection. Large scale, controlled, follow-up studies are currently in progress to confirm these initial findings. PMID- 11391227 TI - Graft size assessment and analysis of donors for living donor liver transplantation using right lobe. AB - BACKGROUND: Modality of living donor liver transplantation (LDLT) has been expanded to adult cases. However, the safety of right lobectomy from living donors has not yet been proven. METHODS: A total of 62 cases of LDLT, using the right lobe, were reviewed. Study 1: Discrepancy between estimated graft volume and actual graft weight was evaluated. Study 2: Postoperative liver functions were analyzed in relation to residual liver volume (RLV) or age. Residual liver volume of donors was defined using two indices, (RLV = estimated whole liver volume - estimated graft volume and %RLV = RLV/estimated whole liver volumex100). Donors were divided into two groups on the basis of either %RLV (<40%; 40%< or =) or age (<50 years old; 50 years old < or =). Study 3: Right lobe donors were compared with left lobe donors (35 cases) in terms of their postoperative liver functions. RESULTS: Study 1: The relationship between estimated graft volume and actual graft weight was linear (y=159.136+0.735x, R2=0.571, P<0.001). Study 2: %RLV ranged from 23.5% to 55.8% (mean +/- SD: 43.2+/-6.0). Fifteen cases showed %RLV less than 40%. Postoperative bilirubin clearance was delayed in that group (%RLV<40%). Serum total bilirubin values on postoperative day 7 in the older group (age > or =50) were significantly higher than those in the younger group (age<50). Study 3: Postoperative liver functions of right lobe donors were significantly higher than those of left-lobe donors. Eleven donors (17.7%) had surgical complications, all of which were cured with proper treatment. CONCLUSIONS: Right lobectomy from living donors is a safe procedure with acceptable morbidity, but some care should be taken early after the operation for donors with small residual liver and aged donors. PMID- 11391228 TI - Bacterial translocation in clinical intestinal transplantation. AB - BACKGROUND: Bacterial translocation (BT) has been suggested to be responsible for the high incidence of infections occurring after small bowel transplantation (SBTx). Bacterial overgrowth, alteration of the mucosal barrier function as a consequence of preservation injury or acute rejection (AR), and potent immunosuppression are all associated with BT. The aim of this study was to evaluate and quantify the correlation of BT with these events. METHODS: Fifty pediatric SBTx recipients on tacrolimus and prednisone immunosuppression were analyzed. Blood, stool, and liver biopsies and peritoneal fluid were cultured (circa 4000 total specimens) when infection was clinically suspected or as part of follow-up. BT episodes were considered when microorganisms were found simultaneously in blood or liver biopsy and stool. RESULTS: BT (average of 2.0 episodes/patient) was evident in 44% of patients and was most frequently caused by Enterococcus, Staphylococcus, Enterobacter, and Klebsiella. The presence of a colon allograft was associated with a higher rate of BT (75% vs. 33.3%). Furthermore, the transplantation procedure (colon vs. no colon) affected the rate of BT: SBTx=40% vs. 25%, combined liver and SBTx=100% vs. 30%, multivisceral transplantation=25% vs. 50%. AR was associated with 39% of BT episodes. BT followed AR in 9.6% of the cases. In 5.2% of the cases, positive blood cultures without stool confirmation of the bacteria were seen. Prolonged cold ischemia time (CIT) affected BT rate significantly (CIT>9 hr 76% vs. CIT<9 hr 20.8%). CONCLUSIONS: This study shows that 1) a substantial percentage of, but not all, BT is associated with AR, 2) the presence of a colon allograft increases the risk for BT, and 3) a long CIT is associated with a high incidence of BT after SBTx. PMID- 11391229 TI - beta-Cell dysfunction rather than insulin resistance is the main contributing factor for the development of postrenal transplantation diabetes mellitus. AB - BACKGROUND: Our study was undertaken to investigate the pathogenesis and possible risk factors for postrenal transplantation diabetes mellitus (PTDM). METHODS: We recruited 114 patients with normal glucose tolerance (NGT) and performed both 75 g oral glucose tolerance tests (OGTT) and short insulin tolerance tests 1 week before and 9-12 months after transplantation. RESULTS: The subjects were classified into three groups by World Health Organization criteria on the basis of OGTT after transplantation: (a) 36 (31.6%) subjects with normal glucose tolerance; (b) 51 (45.7%) subjects with impaired glucose tolerance (IGT); and (c) 27 (23.7%) subjects with postrenal transplantation diabetes mellitus. Dosages of steroid and cyclosporine were equivalent among the three groups. Before transplantation, the fasting and 2-hr plasma glucose and proinsulin/insulin (PI/I) ratios were significantly higher in the IGT and PTDM groups than in the NGT group, but the insulin sensitivity index (ISI) was not significantly different among the three groups. In addition, the area under the curve-insulin on OGTT was significantly lower in the PTDM group than in the NGT group. After transplantation, however, the ISI was increased in all groups. Furthermore, the ISI and PI/I ratios revealed significantly higher values in the PTDM group than in the NGT group after transplantation. CONCLUSIONS: These results revealed that fasting and 2-hr plasma glucose levels, as well as the proinsulin/insulin ratio before transplantation, are both possible indicators of beta-cell dysfunction and may be predictors for the development of PTDM. Furthermore, beta-cell dysfunction, rather than insulin resistance, was proven to be the main factor for the pathogenesis of PTDM. PMID- 11391230 TI - Renal replacement therapy and orthotopic liver transplantation: the role of continuous veno-venous hemodialysis. AB - BACKGROUND: The need for renal replacement therapy (RRT) either before or after orthotopic liver transplant (OLTX) has been reported to be a poor prognostic indicator for survival. Use of continuous veno-venous hemodialysis (CVVHD) for RRT has been reported in three series of OLTX patients with high 90-day mortality rates of 57-60%. We have examined our patient population to determine the effect of necessity and type of RRT on patient survival after OLTX. METHODS: We analyzed 1535 OLTX that were performed at our institution from 1985 through 1999, 1037 from 1985 to 1995 (period I) and 498 from 1996 to 1999 (period II). Combined liver-kidney transplants were excluded from analysis. Hospital dialysis unit records and a prospectively maintained database on all OLTX patients served as the source of data. Patients were classified into groups defined on whether or not they received RRT, when they received RRT, and the type of RRT. Groups were compared for preoperative intensive care unit status, time on the waiting list, laboratory variables, 90-day postoperative mortality, 1-year patient survival, and absolute survival. RESULTS: Use of RRT increased from 8.29% in period I to 12.45% in period II, along with increased median waiting times. In period I, patients receiving preoperative RRT had a 90-day mortality (0%) and a 1-year survival (89.5%) almost identical to those patients who never required RRT (1.7% and 90.6%). Patients who developed acute renal failure postoperatively requiring RRT, however, had a 90-day mortality of 28.6% and a 1-year survival of 55%. In period II, patients requiring RRT had a 90-day mortality of 39.7% and a 1-year actuarial survival of 54.5% compared with 6.9% and 88.6% in patients never requiring RRT. Patients treated with CVVHD had a 90-day mortality of 42% compared with 25% in patients treated with hemodialysis alone. However, patients receiving CVVHD both pre- and postoperatively had a 90-day mortality of 27.7% vs. 50% in those patients who only received CVVHD postoperatively. Patients who developed acute renal failure postoperatively, which required RRT, regardless of therapy, had a 1-year survival of only 41.0% compared with a 1-year survival of 73.6% in those patients started on RRT preoperatively, P=0.03. CONCLUSIONS: The need for RRT has increased along with waiting time in OLTX patients. Patients developing the need for RRT postoperatively have an increased 90-day mortality and lower 1 year survival with the highest being present in patients receiving CVVHD, which was started postoperatively. These findings may reflect a trend toward a sicker population awaiting OLTX and emphasize the negative impact of renal failure on survival after OLTX. PMID- 11391231 TI - Mycophenolate mofetil impairs healing of left-sided colon anastomoses. AB - INTRODUCTION: Inadequate healing and consequent leakage from bowel anastomoses are a significant cause of postoperative morbidity and mortality. Immunosuppressive drugs are known to disturb healing processes and to impair the mechanical stability of bowel anastomosis. Mycophenolate mofetil (MMF) is an immunosuppressive agent that selectively inhibits the proliferation of T and B lymphocytes and has been shown to be effective in preventing allograft rejection after organ transplantation. Adverse effects are few; however, nothing is known in regard to possible adverse effects of MMF administration on the healing of bowel anastomosis. The aim of the present study was to evaluate the effect of systemic MMF administration on the healing of colon anastomoses in rats. METHODS: Rats underwent laparotomy, division of the left colon, and sigmoidostomy. MMF (25 mg/kg) or vehicle was administered intraperitoneally in two groups (n=21 per group) 3 days before surgery and then once daily until euthanization (7 animals per group; 2, 4, and 6 days after surgery). Bursting pressure measurements, histologic evaluation, morphometric analysis, mucin and collagen staining, and BrdU immunohistochemistry of the anastomotic site were performed. Furthermore, matrix protein expression at the anastomotic site was determined by collagen I and fibronectin Western blots. RESULTS: Administration of MMF significantly decreased anastomotic bursting pressure postoperatively. Accordingly, histology, mucin staining, and BrdU immunohistochemistry and measurements of the colonic crypt depth showed more extended inflammation, a significantly lower proliferation rate, and a significantly thinned mucosal layer in the MMF-treated groups when compared to control animals, whereas matrix synthesis at the anastomotic site was not different. CONCLUSIONS: The administration of the immunosuppressive agent MMF significantly impairs healing and mechanical stability of colon anastomosis in rats during the early postoperative period. MMF act to disturb host reparative processes mainly by impairment of reparative colonic epithelium proliferation and less by a disturbance of matrix synthesis. PMID- 11391232 TI - Differential inhibitory effects of intravenous immunoglobulin preparations on HLA alloantibodies in vitro. AB - BACKGROUND: Treatment of allosensitized patients with intravenously administered pooled immunoglobulin preparations (IVIG) may lead to a long-lasting reduction of anti-HLA alloantibody titers. An inhibitory response of IVIG preparations on lymphocytotoxicity is suggested to depend on IgG and to predict a successful reduction of anti-HLA alloantibodies upon the administration of high-dose IVIG in vivo. METHODS: In this study, we evaluated different IVIG preparations for their in vitro inhibitory capacity on lymphocytotoxicity and binding of anti-HLA alloantibodies to purified HLA antigens. For that purpose sera from 24 highly sensitized patients awaiting kidney transplantation and serological HLA testing reagents were used. Panel-reactive antibody (PRA) determinations using standard complement-dependent cytotoxicity testing and anti-HLA alloantibody determination by ELISA were carried out in the presence and absence of 50% (v/v) IVIG. RESULTS: The addition of IgG-containing IVIG preparations gave only a moderate inhibitory response judging from the average decrease of PRA levels (absolute DeltaPRA range: -2% to 16%), whereas the largest inhibition of lymphocytotoxicity was seen after the addition of IgM/IgA-containing IVIG preparations (absolute DeltaPRA range: 19% to 44%). For both IgG and IgM/IgA-containing IVIG preparations, the reduction of lymphocytotoxicity occurred in a dose-dependent fashion without a preference for particular anti-HLA class I antibody specificities. Significantly lower inhibitory effects on anti-HLA antibody reactivity were observed when the effects of IVIG preparations were monitored by ELISA (absolute DeltaPRA range: 7% to 22%). CONCLUSIONS: Our data suggest that the immunomodulatory capacity is largely caused by the IgM/IgA fraction of IVIG when analyzed by lymphocytotoxicity. The different effect on ELISA versus complement-dependent cytotoxicity testing suggests that interactions of IVIG with complement rather than anti-idiotypic antibodies may contribute to the inhibitory effects of IVIG in vitro. PMID- 11391233 TI - Mechanisms of acquired thymic tolerance: induction of transplant tolerance by adoptive transfer of in vivo allomhc peptide activated syngeneic T cells. AB - BACKGROUND: Our most recent observation that i.v. injection of Wistar-Furth (WF) major histocompatibility complex Class I peptide 5 (P5)-pulsed self-myeloid or lymphoid dendritic cells (DC) induces transplantation tolerance suggests that adoptive transfer of in vivo allopeptide-primed host T cells might induce acquired tolerance through their interaction with thymic DC. METHODS: To examine this hypothesis, host myeloid DC cultured in rat granulocyte/macrophage colony stimulating factor and interleukin 4 were pulsed in vitro with P5 and injected intravenously into syngeneic ACI rats. The T cells primed to P5 via the indirect pathway of allorecognition were harvested 7 days later and administered by either intravenously or intrathymically into syngeneic ACI recipients of WF cardiac allografts. RESULTS: Syngeneic T cells obtained from the spleen of P5-primed rats had a high mixed lymphocyte reaction proliferative response to P5 presented by self-DC. I.v. administration of 2x107 P5-primed alloreactive purified host splenic T cells alone on day -7 significantly (P<0.001) prolonged cardiac allograft survival from 10.5+/-1.0 days to 18.6+/-1.8 days in the WF-to-ACI rat combination. I.v. injection of P5-activated host T cells combined with 0.5 ml antilymphocyte serum (ALS)-transient immunosuppression on day -7 led to 100% donor-specific permanent graft survival (>200 days). Thymectomy before i.v. injection of P5-activated syngeneic T cells led to acute graft rejection, suggesting that the homing of in vivo activated T cells to the host thymus might play a role in the induction of tolerance. To further define the role of the recipient thymus in this model, we examined the effects of intrathymic (i.t.) injection of P5-primed alloreactive T cells on graft survival and found that i.t. administration of the P5-primed T cells on day -7 alone significantly prolonged graft survival (15.0+/-0.7 days) and when combined with 0.5 ml ALS led to donor specific permanent graft survival. The long-term unresponsive recipients permanently (>100 days) accepted second-set donor-specific cardiac allografts but not third-party (Lewis) grafts. CONCLUSIONS: These findings demonstrate that the adoptive transfer of splenic T cells primed to an indirectly presented donor peptide induces transplantation tolerance in a transiently immunosuppressed secondary syngeneic recipient. Our data suggest that the interaction of thymic DC with activated peripheral T cells induces alloantigen (Ag)-specific T-cell tolerance by either inactivation or deletion of alloreactive T cells in the thymus. This observation provides the first formal evidence that the interaction between thymic DC and activated peripheral T cells that continuously circulate through the thymus plays an important role in the induction and maintenance of Ag specific tolerance. PMID- 11391234 TI - Generation of helper and cytotoxic CD4+T cell clones specific for the minor histocompatibility antigen H-Y, after in vitro priming of human T cells by HLA identical monocyte-derived dendritic cells. AB - BACKGROUND: There is now convincing evidence that minor histocompatibility antigens (mHag) may play a significant role in the pathogenesis of graft-versus host disease after HLA-identical bone marrow transplantation. Indeed, in this clinical situation, T cells specific for mHag have been isolated. Here, we addressed whether one can generate mHag-specific T cells in vitro, without any in vivo immunization, among healthy blood donors. METHODS: We used monocyte-derived dendritic cells (Mo-DCs) as antigen presenting cells to induce primary responses between healthy HLA-identical siblings, in mixed lymphocyte dendritic cell reactions (MLDCRs). RESULTS: We show that CD4+ T-cell clones, specific for the mHag H-Y, can be generated in vitro. These clones were derived from a gender mismatched positive MLDCR pair of HLA-identical siblings and were restricted by the HLA DQB1*0502 molecule. In addition, these CD4+ T clones were also able to lyse allogeneic targets with the same pattern of restriction and specificity than helper function. Finally, acute myeloid leukemia (AML) blast cells were susceptible to lysis by these clones. CONCLUSIONS: Altogether, these results predict that Mo-DCs could help to generate class II-associated, mHag-specific, T cell lines or clones in vitro, between healthy blood donors, without any need of transplantation-mediated immunization. PMID- 11391235 TI - Microchimeric cells from the peripheral blood associated with cardiac grafts are bone marrow derived, long-lived and maintain acquired tolerance to minor histocompatibility antigen H-Y. AB - BACKGROUND: Although it has been well established that the microchimerism occurs in the peripheral blood of the recipients after various settings in both clinical and experimental organ transplantation, nevertheless, their roles in inducing and maintaining acquired transplantation tolerance are controversial. Furthermore, regarding the cell lineages, kinetics, and functions of the cells that constitute the microchimerism after organ transplantation, solid information is not available. METHODS: Using rat heterotopic heart isografts from bone marrow chimeras between cross-sex and applying polymerase chain reaction with specific primers to rat sex determining region of Y chromosome, a relationship between a state of microchimerism and induction as well as maintenance of acquired tolerance to H-Y antigen were examined. RESULTS: Microchimeric cells of the peripheral blood (MCPB) after cardiac grafting contain bone marrow-derived and radiation-sensitive cells. Furthermore, removal of the primary cardiac grafts revealed that microchimeric cells in the peripheral blood are long-lived cells, i.e., more than 6 months. When the female rats that had contained long-lasting MCPB, were innoculated with syngeneic male dendritic cells, failure to sensitize female toward male specific antigen H-Y was found to occur. CONCLUSIONS: Thus it was suggested that radiation-sensitive, bone marrow derived, long-lived MCPB play a significant role in maintaining acquired transplantation tolerance to minor histocompatibility antigen H-Y. PMID- 11391236 TI - The transforming growth factor-beta1 codon 10 gene polymorphism and accelerated graft vascular disease after clinical heart transplantation. AB - BACKGROUND: The multifunctional cytokine transforming growth factor- (TGF) beta1 is thought to play a role in the pathogenesis of graft vascular disease (GVD). Polymorphisms at codon 10, (Leu10-->Pro) and codon 25 (Arg25-->Pro) in the signal sequence of the TGF-beta1 gene regulate the production and secretion of the protein. We investigated whether these polymorphisms are risk factors for the development of GVD after clinical heart transplantation. METHOD: TGF-beta1 polymorphisms, Leu10-->Pro and Arg25-->Pro, were determined in DNA from heart transplant recipients (n=252) and their donors (n=213), using sequence-specific oligonucleotide probing. GVD was angiographically diagnosed 1 year after transplantation. In addition other potential risk factors including underlying disease, recipient and donor age, recipient and donor gender, number of acute rejections in the first year, cold ischemia time, and HLA mismatches were analyzed by univariate and multivariate logistic regression analysis. RESULTS: Univariate analysis showed that the recipient TGF-beta1 polymorphism Leu10-->Pro, (P=0.056, chi2 test), underlying disease (P=0.01, chi2 test), number of acute rejections in the first-year (P=0.03, analysis of variance), and donor age (P<0.001, analysis of variance) were risk factors for the development of GVD. The TGF-beta1 Arg25-->Pro polymorphism was not a risk factor. Also in the multivariate analysis, the recipient TGF-beta1 codon 10 polymorphism was associated with GVD, with patients homozygous for Pro at greatest risk (odds ratio 7.7, P=0.03). Apart for the recipient TGF-beta1 Leu10-->Pro polymorphism, donor age appeared to be an independent risk factor for the development of GVD at 1 year. Patients with older donor hearts were at greater risk than patients receiving grafts from younger donors (odds ratio 1.1/year, P<0.001). CONCLUSION: Recipient TGF-beta1 Leu10-->Pro polymorphism and higher donor age are independent risk factors for the development of GVD after clinical heart transplantation. PMID- 11391237 TI - In vitro analysis of CD40-CD154 and CD28-CD80/86 interactions in the primary T cell response to allogeneic "nonprofessional" antigen presenting cells. AB - BACKGROUND: Recently, several ligand interactions have been examined in detail as potential mediators of costimulatory signaling. The CD154/CD40 and CD28/B7 interactions have been highlighted as being among the more-significant contributors to proper activation of unprimed T lymphocytes. Human keratinocytes (HK) and human dermal fibroblasts (HF) are capable of expressing Class II HLA and CD40 antigens after interferon-gamma exposure, yet neither express significant levels of B7. HK and HF have been characterized as "nonprofessional" antigen presenting cells (APC) and their poor APC function has been partially attributed to deficient costimulatory activity. METHODS: In this study, we examined whether substituting for costimulatory signaling events through the addition of cross linked monoclonal antibodies against the T-cell ligand/s (CD28 and/or CD154) could restore allostimulation. Mixed lymphocyte reactions were performed combining enriched human peripheral blood T cells and allogeneic HK or HF with or without stimulatory anti-CD28 and/or anti-CD154 antibodies. RESULTS: The results show that the addition of anti-CD28 alone permitted HF but not HK to present alloantigen effectively. In contrast, addition of both anti-CD154 and anti-CD28 was required to generate even a moderate proliferative response to allogeneic HK. Further, adding a monomorphic anti-HLA-DR antibody substantially inhibited these responses. Additional experiments suggest that signaling through CD40/CD154 directs HK to produce TGF-beta, which would adversely affect T-cell activation. CONCLUSIONS: The data presented highlight significant differences in signaling capacities for HK versus HF and provide evidence for a partial mechanism by which allogeneic human skin equivalents might be immunologically null upon engraftment. PMID- 11391238 TI - Role of cytokine gene polymorphism in hepatitis C recurrence and allograft rejection among liver transplant recipients. AB - BACKGROUND: Cytokines play a key role in the regulation of immune responses. The maximal capacity of cytokine production varies between individuals and was shown to correlate with polymorphism in cytokine gene promoters. The objective of this study was to analyze the role of cytokine allelic variations in susceptibility to early graft rejection episodes and recurrence of hepatitis C infection in liver transplant (LTx) recipients. METHODS: The genetic profile of five cytokines was studied in 68 LTx recipients and 49 controls using polymerase chain reaction sequence specific primers. All individuals were genotyped as high or low producers of TNF-alpha and IL-6 and high, intermediate, or low producers of transforming growth factor beta (TGF-beta), interferon gamma (IFN-gamma), and interleukin 10 (IL-10) based on single nucleotide substitutions. RESULTS: No statistically significant differences were observed between patients with or without early rejection episodes. A significant proportion of patients more prone to rejection were genotyped as having a low production profile of IL-10 compared with the control population (P=0.04). These data are in accordance with reports regarding other solid-organ transplant recipients. Patients with no recurrence of hepatitis C had the inherent ability to produce higher TGF-beta levels than did patients with recurrent disease (P=0.042). Among nonrecurrent patients, the percentage of genetically low IL-10 producers was higher than among recurrent patients (P=0.07). Furthermore, a genetic tendency to produce higher levels of IFN-gamma was noted among LTx recipients with nonrecurrent hepatitis C than among those with recurrent hepatitis C. CONCLUSIONS: While no significant correlation was detected between particular cytokine profile and early rejection episodes, our data strongly suggest an association between cytokine gene polymorphism of TGF-beta, IL-10, and INF-gamma and recurrence of hepatitis C in LTx recipients. PMID- 11391239 TI - Screening for solid organ malignancies prior to heart transplantation. AB - BACKGROUND: Prognosis of solid organ cancer in immunosuppressed hosts is generally dismal. Therefore, every effort to identify patients with asymptomatic carcinomas before transplantation should be encouraged. METHODS: Sixty-seven patients referred for heart transplantation were examined adhering to the scheme proposed at the 24th Bethesda Conference. To increase the sensitivity of this work-up, the following items were added: tumor marker assays (prostate-specific antigen in males, carcino embryogenic antigen), abdominal ultrasound, CT scan of the abdomen and the thorax, mammography/echography of the breasts, PAP smear, colonoscopy if carcino embryogenic antigen abnormal or occult blood in stool, prostate echography if prostate-specific antigen abnormal or prostate hypertrophy. RESULTS: Carcinoma was detected in 10 of the 67 patients; for 8 patients of this cancer group, transplantation was denied. Importantly, 9 of the 10 malignancies were detected by means of the diagnostic items that were added to the standard screening protocol. There were no significant differences between the cancer and the non-cancer group regarding mean age, sex, etiology of heart failure, and smoking history. Stratifying patients in younger (i.e., < or =54 years) and older (i.e., > or =55 years) age groups showed a significantly greater proportion of older patients in the cancer group (8/10=80%) compared to the non cancer group (25/57=44%), P=0.04. After a mean follow-up of 34 months, 5 of the 36 transplanted patients developed a malignancy (4 skin carcinomas, 1 non-Hodgkin lymphoma). There have been no malignancy-related deaths until now. CONCLUSION: The importance of a thorough screening program in the triage of candidates with preexisting malignancies, especially in an older patient population, is illustrated in this report. PMID- 11391240 TI - Late onset of severe graft-versus-host disease in a pediatric liver transplant recipient. AB - We report the management of a patient with the late onset of chronic graft-versus host disease (GVHD) after orthotopic liver transplantation. GVHD is a rare complication of solid organ transplants that usually presents early after transplantation and is fatal in the majority of cases. Our patient differs from the typical patient with GVHD in that the onset of her disease was very late. Although most treatment to date consisted of an increase in immunosuppressive therapy, our patient showed an excellent response to a reduction. This resulted in the abatement of the symptoms of GVHD and the preservation of her allograft function. PMID- 11391242 TI - The diagnostic challenge of pulmonary Kaposi's sarcoma with pulmonary tuberculosis in a renal transplant recipient: a case report. AB - We report a case of a 39-year-old, HIV-negative, post renal transplant patient who developed mucocutaneous Kaposi's sarcoma with lung parenchymal involvement and concurrently culture proven pulmonary tuberculosis. To the best of our knowledge, this is the first case report of this combination, which presented with cavitating lung nodules and responded well to withdrawal of immunosuppressive drugs beside antituberculous treatment. PMID- 11391241 TI - Interferon alpha 2B and ribavirin in severe recurrent cholestatic hepatitis C. AB - Severe recurrent cholestatic hepatitis C after liver transplantation has a poor prognosis and no standard therapy is currently available. Four cases of severe recurrent cholestatic hepatitis C treated with a combination of interferon alpha 2b and ribavirin are described. All four patients were transplanted for hepatitis C-related cirrhosis. The mean age at transplantation was 45 years (range 41-51 years). Three of the patients were male and one was female. All four patients had hepatitis C virus viremia before and after liver transplantation. At 2 to 23 months after liver transplantation, all four patients developed jaundice, cholestatic elevation of liver enzymes, and histopathology consistent with severe recurrent cholestatic hepatitis C. Combination of interferon and ribavirin was given with prompt virological suppression. Despite this rapid viral suppression, all four patients developed progressive graft failure with three deaths. PMID- 11391243 TI - Postnatal cytokines and boosts improve chimerism and hematological parameters in beta-thalassemic mice transplanted in utero. AB - We have developed a murine model of in utero transplantation in nonanemic, beta thalassemic mice to study chimerism, tolerance, and changes in hematological parameters in response to cytokines and postnatal boosts with donor cells. We have documented improved survival of homozygous fetuses by 40% as compared with controls. Low-level, mixed chimerism was improved by postnatal cytokine therapy and boosts and was associated with improvement in hemoglobin levels, reticulocyte counts, and iron stores. Cytotoxicity assays demonstrated higher responses to donor cells in control mice as compared with in utero transplanted animals (at 50:1 effector to target ratios, transplanted mice showed 8.66% target lysis and control mice showed 51.85% target lysis, P=0.0003), indicating tolerance. The combination of prenatal tolerance to allogeneic cells with postnatal boosts in primed hosts may become an effective, nontoxic strategy for the improvement of hemolytic anemia in beta-thalassemic patients. PMID- 11391244 TI - A pediatric patient with classical citrullinemia who underwent living-related partial liver transplantation. AB - Patients with inborn errors of metabolism undergo liver transplantation, but the effect of transplanting the liver of healthy carriers of these conditions has not been fully clarified. A 6-year-old girl with classical citrullinemia, who repeatedly suffered from hyperammonemia, underwent living-related liver transplantation by using a segment of the liver of her mother, who was a heterozygote carrier for classical citrullinemia. Hyperammonemia alleviated in the patient after the transplantation, thereby dramatically improving her quality of life. Although the levels of plasma and urinary citrulline remained high postoperatively, there was no marked difference in the level of plasma citrulline up to 1 month after surgery when compared with that of previously reported orthotopic liver transplantation cases with classical citrullinemia. PMID- 11391245 TI - Emergency adult to adult living donor liver transplantation for fulminant hepatic failure--is it justifiable? PMID- 11391247 TI - Sex, the heart, and sildenafil. PMID- 11391248 TI - Diagnostic laryngeal electromyography: The Wake Forest experience 1995-1999. AB - BACKGROUND: Laryngeal electromyography (LEMG) is a valuable diagnostic/prognostic test for patients with suspected laryngeal neuromuscular disorders. OBJECTIVE: To report our experience with diagnostic LEMG at the Center for Voice Disorders of Wake Forest University and to evaluate the impact of LEMG on clinical management. METHODS: Retrospective chart review of 415 patients who underwent diagnostic LEMG over a 5-year period (1995-1999). RESULTS: Of 415 studies, 83% (346 of 415) were abnormal, indicating a neuropathic process. LEMG results altered the diagnostic evaluation (eg, the type of radiographic imaging) in 11% (46 of 415) of the patients. Unexpected LEMG findings (eg, contralateral neuropathy) were found in 26% (107 of 415) of the patients, and LEMG results differentiated vocal fold paralysis from fixation in 12% (49 of 415). Finally, LEMG results altered the clinical management (eg, changed the timing and/or type of surgical procedure) in 40% (166 of 415) of the patients. CONCLUSIONS: LEMG is a valuable diagnostic test that aids the clinician in the diagnosis and management of laryngeal neuromuscular disorders. PMID- 11391249 TI - The importance of hyaluronic acid in vocal fold biomechanics. AB - OBJECTIVE: This study examined the influence of hyaluronic acid (HA) on the biomechanical properties of the human vocal fold cover (the superficial layer of the lamina propria). STUDY DESIGN: Vocal fold tissues were freshly excised from 5 adult male cadavers and were treated with bovine testicular hyaluronidase to selectively remove HA from the lamina propria extracellular matrix (ECM). Linear viscoelastic shear properties (elastic shear modulus and dynamic viscosity) of the tissue samples before and after enzymatic treatment were quantified as a function of frequency (0.01 to 15 Hz) by a parallel-plate rotational rheometer at 37 degrees C. RESULTS: On removing HA from the vocal fold ECM, the elastic shear modulus (G' ) or stiffness of the vocal fold cover decreased by an average of around 35%, while the dynamic viscosity (eta') increased by 70% at higher frequencies (>1 Hz). CONCLUSION: The results suggested that HA plays an important role in determining the biomechanical properties of the vocal fold cover. As a highly hydrated glycosaminoglycan in the vocal fold ECM, it likely contributes to the maintenance of an optimal tissue viscosity that may facilitate phonation, and an optimal tissue stiffness that may be important for vocal fundamental frequency control. SIGNIFICANCE: HA has been proposed as a potential bioimplant for the surgical repair of vocal fold ECM defects (eg, vocal fold scarring and sulcus vocalis). Our results suggested that such clinical use may be potentially optimal for voice production from a biomechanical perspective. PMID- 11391250 TI - Failed medialization laryngoplasty: management by revision surgery. AB - OBJECTIVE: The purpose of this study was to evaluate the cause of immediate and late medialization laryngoplasty failures and to describe their management. METHODS: A retrospective analysis was performed in 20 patients who underwent revision surgery after failed medialization laryngoplasty. Analysis was based on preoperative spiral CT scan, preoperative and postoperative videostrobolaryngoscopy, and phonatory function measures. RESULTS: Three major types of failures were identified. The most common problem was arytenoid rotation with a persistent posterior glottic gap (11 of 20). Malposition or wrong size of the implants resulted in a lateralized vocal fold or false vocal fold medialization (6 of 20). Three patients had implants that were extruding. Late atrophy and bowing resulted in a glottal gap (2 of 20). One patient had fibrosis around the implant requiring removal. Spiral CT scan of the larynx located the implant precisely and showed the degree of arytenoid rotation. Patients with arytenoid rotation and posterior gap had revision medialization combined with arytenoid adduction. Revision medialization was performed in 11 patients, arytenoid adduction in 12 patients, lipoinjection in 2 patients, and 4 implants were removed. The voice was improved in 15 patients. Improved voice was correlated with improved phonation time and reduced phonatory airflow rates. CONCLUSION: Immediate and late failures of medialization laryngoplasty are due to several possible causes. Revision surgery is feasible and highly successful. To select between the surgical alternatives work up should include preoperative analysis of vocal function, videostrobolaryngoscopic analysis, and spiral CT of the larynx. PMID- 11391251 TI - Laryngopharyngeal sensory deficits as a predictor of aspiration. AB - OBJECTIVE: This study evaluates whether patients with severe sensory deficits in the hypopharynx are at increased risk for aspiration and determines the relationship between pharyngeal muscular weakness and hypopharyngeal sensory deficits. STUDY DESIGN AND SETTING: Forty patients with dysphagia who underwent flexible endoscopic evaluation of swallowing with sensory testing were prospectively divided into 2 groups. One group included patients with severe sensory deficits determined by an absent laryngeal adductor reflex and the other with normal sensitivity. Subjects were given liquid and puree consistencies and were evaluated for aspiration as well as pharyngeal muscle contraction. RESULTS: The differences in incidence of aspiration and pharyngeal muscular weakness between the 2 groups were significant (P < 0.001 Fisher's exact test). CONCLUSION: There is a strong association between motor function deficits and hypopharyngeal sensory deficits. SIGNIFICANCE: The association of sensory loss and motor deficits together with the use of flexible endoscopic evaluation of swallowing with sensory testing can predict those patients who are at highest risk for aspiration. PMID- 11391252 TI - Postintubation phonatory insufficiency: an elusive diagnosis. AB - OBJECTIVE: To describe postintubation phonatory insufficiency, a routinely overlooked complication of prolonged intubation. This entity results from an erosive injury with permanent divots of the medial surfaces of the arytenoid cartilages and/or scarring of the anterior cricoarytenoid joint capsule so that during phonation there is incomplete adduction and a glottic gap remains. SETTING: Tertiary care center. STUDY DESIGN/RESULTS: We present a retrospective review of the findings of 138 patients evaluated for chronic intubation injuries in our voice laboratory using a diagnostic model composed of pertinent history, elicited vocal capabilities and limitations, and an intense fiberoptic laryngeal examination directed at revealing the suspected injuries. CONCLUSION/SIGNIFICANCE: In many patients, the diagnosis of postintubation phonatory insufficiency was made years after the intubation injury occurred and after numerous nondiagnostic examinations elsewhere. Mechanisms of intubation injury are reviewed, and prevention and treatment of the condition are discussed. PMID- 11391253 TI - Long-term follow-up of cartilaginous tumors of the larynx. AB - BACKGROUND: Cartilaginous tumors of the larynx are uncommon. A literature review disclosed approximately 250 cases since 1816; the cricoid cartilage is the most common site. The rarity of these tumors has made for limited experience and, as a consequence, our knowledge is incomplete. OBJECTIVE: To report surgical results as well as long-term follow-up on 6 patients with cartilaginous tumors of the larynx. DESIGN: A 28-year retrospective study with the patients followed-up from 6 to 28 years (average, 17.8 years). METHODS: Six adult white male patients with cartilaginous tumors of the larynx: 4 low-grade chondrosarcoma (1 of the thyroid and 3 of the cricoid) and 2 chondroma of the cricoid. Surgical treatment included total laryngectomy of the thyroid and 1 of the cricoid chondrosarcoma, and conservation surgery of the other 4 cricoid tumors: the 2 patients with chondrosarcoma had total resection of the cricoid cartilage with thyrotracheal anastomosis, and the 2 patients with chondroma had local tumor resection using a laryngofissure approach. RESULTS: The margins of the specimen were negative for tumor in the 6 patients. On follow-up, none of the patients had regional or distant metastasis or tumor-related death. One of the patients with cricoid chondrosarcoma developed recurrence 8 years after conservation surgery, and required a total laryngectomy for salvage. Survival rate tumor-free at 5 years was 100% and at 10 years 67%, co-morbidity being responsible for the decrease in survival rate. CONCLUSIONS: Based on this small series of patients, the long-term follow-up of benign and low-grade malignant tumors suggests that the surgical approach and prognosis does not depend on histologic distinction and, importantly, underdiagnosed malignancy on tumor sampling and recurrent chondrosarcoma, managed with salvage surgery, have no adverse impact on patient survival. Total resection of the cricoid cartilage with thyrotracheal anastomosis over a stent proved an alternative surgical technique in chondrosarcoma who otherwise would have been treated by total laryngectomy. PMID- 11391254 TI - Duration of neuronal stretch correlates with functional loss. AB - Postoperative cranial nerve weakness or paralysis is not uncommon in many otolaryngologic surgical procedures. Our study used a rat model to test the hypothesis that the length of time that a nerve is under tension may be an important variable in the amount of postoperative paresis. Forty Sprague-Dawley rats were divided into 4 groups that underwent either a sham operation or a traction injury for 1, 2, or 5 minutes. The traction injury was performed with a vessel loop placed around the sciatic nerve with 50 g of tension. Traction injury for 1 or 2 minutes did not result in any statistical differences in the motor capabilities of the lower limb. However, those animals with a stretch injury for 5 minutes had a significant loss of function (P < 0.01) when compared with all other groups. Histologic examination of nerves harvested on postoperative day 7 showed no evidence of mechanical injury. This study demonstrates that even minimal tension, if maintained for a significant amount of time, may result in postoperative weakness. PMID- 11391255 TI - Long-term results of the first 500 cases of acoustic neuroma surgery. AB - OBJECTIVE: This retrospective study focuses on 2 outcome results after surgical intervention for acoustic neuroma: (1) facial nerve status, and (2) hearing preservation. STUDY DESIGN: A total of 484 patients with an acoustic neuroma. RESULTS: Postoperative facial nerve outcomes were significantly different (P < 0.001) according to the size of the tumors. Tumor size had even more influence on the immediate postoperative results. In addition, statistical significance (P < 0.05) was demonstrated in comparing facial nerve outcomes with the surgeon's surgical experience. We also noted that as the patient's age increases, the likelihood for facial dysfunction may increase for all postoperative intervals. The overall success rate of retaining useful hearing was 27% (26 of 95). Class A hearing was retained in 66% (10 of 15) of cases operated on through middle fossa approach in the last 5 years. CONCLUSION: This study demonstrates that tumor size and surgeon's experience are the most significant factors influencing the facial nerve status and hearing outcome after removal of acoustic neuroma. PMID- 11391256 TI - Magnetic resonance imaging in the search for retrocochlear pathology. AB - OBJECTIVES: The purpose of this study is to evaluate the incidence of retrocochlear pathologic conditions that may be seen with full, contrast-enhanced magnetic resonance imaging of the brain, relative to more limited studies of the internal auditory canals (IACs) in patients with unilateral hearing loss or tinnitus with or without dizziness. METHODS: A retrospective study was performed on all patients who had a magnetic resonance imaging of the brain and otologic symptoms over a 7-year period. RESULTS: Abnormalities were found in 11 of 128 patients with only auditory symptoms and in 63 of 282 patients with auditory symptoms and/or dizziness (P = 0.005). Treatable lesions may not have been identified with a limited IAC study in 3 of 128 patients with only unilateral auditory symptoms, and 21 of 282 patients with dizziness (P = 0.045). CONCLUSIONS: Limited MRI of the IACs may serve as an adequate screening test for retrocochlear pathologic conditions in patients with unilateral auditory symptoms and no dizziness. PMID- 11391257 TI - Third place--Resident Research Competition, AAO-2000. Antisense cyclin D1 inhibits growth of head and neck cancer xenografts in nude mice. AB - PROBLEM: Cyclin D1 is a regulatory factor essential in the progression of the cell cycle from G1 through S phase. Amplification and overexpression of cyclin D1 have been observed in many human cancers including head and neck squamous cell carcinoma (HNSCC). We have previously transfected a HNSCC control cell line (CCL23) with an antisense cyclin D1 plasmid and demonstrated inhibition of cell proliferation in vitro. In this study, we examine whether antisense cyclin D1 could inhibit tumor growth in vivo. Methods/measures: The CCL23 and its antisense cyclin D1 transfected clone (CCL23 AS) were injected into the flanks of nude mice. Tumor growth was monitored weekly. After 5 weeks, tumors were removed and studied for tumor size, cyclin D1 expression, cyclin D1-dependent kinase activity, and retinoblastoma (Rb) phosphorylation. RESULTS: Compared with the control tumors, 11 of 19 antisense tumors were smaller, 7 tumors were of equal size, and 1 tumor was larger. Immunohistochemical analysis with an anti-cyclin D1 antibody demonstrated decreased cyclin D1 expression in CCL23 AS and the smaller antisense tumors. Cyclin D1-dependent kinase activity was reduced in CCL23 AS and the smaller antisense tumors, and this was accompanied by a relative decrease in phosphorylated Rb in these samples. CONCLUSION: Antisense cyclin D1 inhibits growth of HNSCC tumors. Cyclin D1 expression, cyclin D1-dependent kinase activity, and Rb phosphorylation are decreased in these tumors. CLINICAL SIGNIFICANCE: These findings lend support for the potential use of antisense cyclin D1 as gene therapy for HNSCC. PMID- 11391258 TI - Identification of differentially expressed genes in human papillomavirus type-16 infected oral cancer cells. AB - OBJECTIVE: The goal of this study was to determine the genes required for head and neck cancer development. STUDY DESIGN AND SETTING: Differential mRNA display analysis was performed using human papillomavirus Type-16 infected immortalized human oral keratinocytes (HOK-16B) and its benzo(a)pyrene-exposed tumorigenic derivative (HOK-16B-BaP-T). RESULTS: Twenty-one differentially expressed cDNA clones were identified between the 2 cell lines. Clone 4 with no known homology showed lower expression in tumorigenic cells compared with either normal or immortalized oral keratinocytes. Clone 6 expression was elevated in several head and neck cancer cells, in addition to Burkitt's lymphoma Raji harboring latent Epstein-Barr virus. CONCLUSION: These findings suggested that clone 6 may be involved in the oncogenic transformation whereas clone 4 may potentially function as a tumor suppressor gene. SIGNIFICANCE: Differential mRNA analysis using the in vitro oral carcinogenesis model may help to identify important genetic markers for the early detection and progression of head and neck cancer. PMID- 11391259 TI - Pentoxifylline effects on acute and late complications after radiotherapy in rabbit. AB - Soft tissue damages after radiotherapy are an uncommon but serious complication. Late damage after radiation is the principal dose-limiting factor in radiation therapy today and is dependent on vascular pathology as a result of radiation. Pentoxifylline is a methylxanthine derivative that produces dose-related improvement in blood flow, lower blood viscosity, improved erythrocyte flexibility, and increased tissue oxygen levels. An agent that increases blood flow and tissue oxygen content may contribute to enhanced healing of soft tissue pathology. Sixteen adult New Zealand rabbits were separated into 2 groups and inspected for 30 weeks after radiation. We noted acute and chronic reactions and pathologic changes in different regions of the head and neck of rabbits. The prophylactic administration of pentoxifylline in the postirradiation period can reduce late soft tissue pathology, but it does not affect acute radiation reactions. PMID- 11391260 TI - Transnasal mucosal flap rotation technique for repair of posterior choanal atresia. AB - PURPOSE: To describe improved surgical treatment for posterior choanal atresia (PCA) by creating mucosal flaps with the aid of operating microscope, CO(2) laser, self-retaining nasal retractor, and stenting with a flat polytetrafluoroethylene (Teflon) keel. MATERIALS AND METHODS: Retrospective study of 19 patients with PCA, their ages ranging from 6 days to 4 years 4 months at the time of first repair, representing a total of 32 PCA repairs. Four patients had unilateral PCA, and one did not return for follow-up and could not be located. A transnasal microscopic approach uses a myringotomy knife or CO(2) laser to create an anterior mucosal flap; the CO(2) laser is also used to remove any bone plate and to create the posterior flap. The flaps then are rotated into position and are kept separated by a flat Teflon keel instead of a traditional round stent to avoid pressure necrosis. When a stable epithelialized opening (2 x 2 to 3 x 4 mm) is created, it is enlarged by subsequent staged transnasal CO(2) laser submucosal scar excisions with preservation of the overlying mucosa. This creates rotation or sliding flaps to speed healing and prevents circumferential scar contraction. RESULTS: Eighteen patients were re-examined after periods of 3 months to more than 5 years from when their first epithelialized orifice was created with nasal endoscopy to measure the final orifice size. All 18 patients had clinically adequate-to-good bilateral nasal airways at the latest follow-up with an average orifice size of 3 x 5 mm and a range of 2 x 2 to 4 x 10 mm. This "same day surgery" technique with staged procedures provides similar or better patency rates than a single transpalatal approach but with less surgical morbidity. It has a much lower restenosis rate than the transnasal puncture or PCA excision with prolonged round tube stenting and multiple dilation technique. PMID- 11391261 TI - Bacteriology of chronic sinusitis after amoxicillin-clavulanate potassium therapy. AB - The bacteriology of chronic sinusitis was studied after amoxicillin-clavulanate potassium therapy. Patients with chronic sinusitis were randomly divided into 2 groups. In the study group, 90 patients were given a 2-week course of amoxicillin clavulanate potassium before functional endoscopic sinus surgery. In the control group, 113 patients did not take any antibiotics within 2 weeks before the surgery. Swab specimens were taken from the maxillary and ethmoid sinuses during surgery and sent for aerobic and anaerobic culture. In the study group, the culture rates of maxillary and ethmoid sinuses were 45.6% and 28.9%, respectively. In the control group, the culture rates of maxillary and ethmoid sinuses were 53.1% and 34.5%, respectively. The culture rates between the control group and the study group were not significantly different, either for the maxillary sinus or the ethmoid sinus. This showed that treatment with amoxicillin clavulanate potassium did not change the bacteriology of chronic sinusitis. PMID- 11391262 TI - Use of AlloDerm as a barrier to treat chronic Frey's syndrome. PMID- 11391263 TI - Endoscopic repair of tension pneumocephalus. PMID- 11391264 TI - Endoscopic surgery for the removal of a nasal glioma: case report. PMID- 11391265 TI - Endodermal sinus tumor of the maxillary sinus. PMID- 11391266 TI - Late-occurring infections of medialization laryngoplasty implants. PMID- 11391267 TI - Bilateral cricoarytenoid joint ankylosis in scleroderma. PMID- 11391268 TI - Foreign body aspiration: an unusual presentation of myasthenia gravis. PMID- 11391269 TI - Acute necrotizing epiglottitis resulting in necrotizing fasciitis of the neck and chest. PMID- 11391270 TI - Ganglioneuroma of the parapharyngeal space in a pediatric patient. PMID- 11391271 TI - Cartilaginous choristoma of the neck associated with a branchial cleft cyst. PMID- 11391272 TI - Hemangioma of the pyriform sinus. PMID- 11391273 TI - Reduction in colonization and nosocomial infection by multiresistant bacteria in a neonatal unit after institution of educational measures and restriction in the use of cephalosporins. AB - INTRODUCTION: Previous administration of third-generation cephalosporins predisposes to colonization and infections by multiresistant Enterobacter sp. The emergence of multiresistant bacteria infections in a neonatal unit during 1995, especially Enterobacter cloacae, stimulated this study. OBJECTIVE: To evaluate the efficacy of measures to control colonization and nosocomial infection by multiresistant bacteria in a neonatal unit. SETTING: A tertiary care university hospital. PATIENTS AND METHODS: This study was conducted from October 1995 through December 1999 in 4 phases: a cross-sectional study, a longitudinal study with intervention measures, monthly cross-sectional studies, and determination of nosocomial infections caused by multiresistant bacteria (oxacillin-resistant Staphylococcus aureus and gram-negative bacteria resistant to either aminoglycosides or third-generation cephalosporins). Specimens for surveillance culture were obtained through umbilical and rectal swabs, and tracheal aspirates from intubated babies. The intervention measures were as follows: (1) appropriated training of the whole health care team, emphasizing measures to reduce cross-colonization, and the importance of rational usage of antibiotics and (2) suppression of usage of third-generation cephalosporins. Risk factors were analyzed through univariate and multivariate logistic regression. RESULTS: In the first phase, 32% (10/31) of the patients were colonized by multiresistant bacteria (29% by multiresistant E cloacae ). In the second phase, 342 patients were evaluated; 33% of them were colonized by E cloacae, and a multiresistant strain was isolated in 10.8% (37/342) of the babies. A logistic regression model indicated parenteral nutrition and antibiotic usage as risk factors for colonization by multiresistant E cloacae. In the third phase, for 6 months, only 2 patients were colonized by multiresistant E cloacae. In the fourth phase, the analysis of bacterial resistance profile indicated a reduction of nosocomial infections due to multiresistant bacteria from 18 cases in 1995 to 2 cases per year until 1999. CONCLUSION: These results have shown that the measures adopted were effective. PMID- 11391274 TI - Antibiotic-resistant organisms among long-term care facility residents on admission to an inpatient geriatrics unit: Retrospective and prospective surveillance. AB - BACKGROUND: There is limited information about antibiotic-resistant organisms in community long-term care facilities (LTCFs). The objective of this study was to obtain data on resistant organisms in residents from community LTCFs admitted to an inpatient acute geriatrics service (AGS). METHODS: Two studies were performed. In the first study, bacteriology records of all admissions to the AGS for the period from November 1, 1998, through June 30, 2000, were reviewed for resistant organisms (methicillin-resistant Staphylococcus aureus [MRSA], vancomycin resistant enterococci [VRE], and resistant gram-negative bacilli). In the second study, residents admitted to the AGS during a 2-month period (N = 92 admissions) had surveillance cultures (nares, gastrostomy site, wounds, and urine) for resistant organisms done within 72 hours of admission. RESULTS: In the retrospective study, there were 727 admissions, of which 437 (60%) had 928 cultures within 72 hours of admission; 590 (64%) cultures grew 1 or more pathogens. Urine (65%) and blood (26%) cultures accounted for 91% of all cultures done. Rates of resistance by culture site were as follows: urine (resistant organism in 16.6% of 373 cultures), blood (6.7% of 60 cultures), wound (52% of 23 cultures), and sputum (40% of 20 cultures). MRSA and enterococci with high-level gentamicin resistance were the most common resistant organisms identified. No VRE were isolated; only 3% of 421 gram-negative isolates were considered resistant strains compared with 19% (P <.001) of gram-positive isolates. In the prospective study, 17% of 92 residents were found to have a resistant organism in 1 or more surveillance cultures; the most common resistant organisms were MRSA and high level gentamicin-resistant enterococci. Only 1 resident was found to have VRE in a rectal swab culture; resistant gram-negative bacilli also were uncommon. CONCLUSIONS: Among residents of community LTCFs admitted to an AGS, resistant organisms were identified infrequently (<20% of admissions). MRSA was the most common resistant organism; VRE and resistant gram-negative bacilli were rare. These findings vary from other studies suggesting that there may be geographic variation in the epidemiology of resistant organisms among residents of community LTCFs. PMID- 11391275 TI - Infection control programs at children's hospitals: a description of structures and processes. AB - BACKGROUND: Infection control (IC) structures and processes determine the effectiveness of surveillance efforts to prevent infections in health care settings. METHODS: A survey was sent to 56 children's hospitals collaborating in the Pediatric Prevention Network (PPN). RESULTS: Completed surveys were returned from 48 hospitals. Responsibility for the IC program resided with the medical director (21%); vice president for patient care (18%); quality improvement director (17%); other senior hospital administrator (15%); or other hospital personnel (18%). Forty-two hospitals had an IC committee; 32 had antimicrobial restriction/control policies; and 21 had an antimicrobial restriction/control task force or committee. Components of antimicrobial restriction programs included infectious disease specialist approval, restricted formularies, selective susceptibility test reporting, and staff education programs. Many methods were used to detect infections, including microbiology laboratory reports (100%); record reviews (98%); informal reports from providers (90%); and readmission reviews (77%). CONCLUSIONS: Children's hospitals vary widely in how they design and implement their IC functions. These variations influence adverse event detection and nosocomial infection rate calculations. If medical errors, including nosocomial infections, are to be detected and hospital rates compared, standardized methods to collect, analyze, and report data are needed. The PPN has initiated activities to standardize surveillance and IC practices in participating hospitals. PMID- 11391277 TI - Pediatric Prevention Network: a multicenter collaboration to improve health care outcomes. AB - Nosocomial infections and antimicrobial resistance are major causes of mortality and morbidity and have become a major public health focus. To date, most national and international nosocomial infection surveillance and prevention activities have been focused on adults, despite the fact that pediatric patients are at high risk for nosocomial infections because of their immature immune systems and prevalent device usage. In 1997 the Hospital Infections Program at the Centers for Disease Control and Prevention and the National Association of Children's Hospitals and Related Institutions partnered to establish a Pediatric Prevention Network. Infection control professionals and their hospital administrators at all children's hospitals were invited to participate. The objectives of the network are to establish baseline infection rates; design, implement, and evaluate prevention interventions; establish benchmark rates and best practices; and serve as a site for multicenter studies to improve outcomes for hospitalized children. This network serves as a model for quality improvement systems in health care. PMID- 11391276 TI - Nosocomial infection rates in US children's hospitals' neonatal and pediatric intensive care units. AB - BACKGROUND: Few data are available on nosocomial infections (NIs) in US children's hospitals' neonatal or pediatric intensive care units. The Pediatric Prevention Network (PPN) was established to improve characterization of NIs in pediatric patients and to develop and test interventions to decrease NI. METHODS: Fifty participating children's hospitals were surveyed in 1998 to determine NI surveillance methods used and neonatal intensive care unit (NICU) and pediatric intensive care unit (PICU) 1997 NI rates. Data were collected on standardized forms and entered and analyzed by using SPSS for Windows. RESULTS: Forty-three (86%) children's hospitals returned a completed questionnaire. All reported conducting NICU and PICU NI surveillance (range, 2-12; median, 12 months). Nineteen children's hospitals provided NICU NI rate data in one or more formats suitable for comparison. Denominators used for NICU NI rate calculations varied: 17 reported overall NI by patient-days; 19 reported bloodstream infection (BSI) by central venous catheter (CVC)-days, and 8 reported BSI by patient-days. Sixteen (16) children's hospitals reported NICU BSI data stratified by CVC-days and birth-weight cohort, and ventilator-associated pneumonia (VAP) by birth weight cohort was reported by 12. Twenty-four children's hospitals reported PICU NI rate data in one or more formats suitable for comparison. Denominators used for PICU NI rate calculations also varied: 20 reported overall NI rates by patient-days; 23 reported BSI rates by CVC-days, and 10 reported BSI rates by patient-days; 24 reported VAP by ventilator-days; and 15 reported urinary tract infections (UTIs) by urinary catheter-days. Median overall NI rates per 1000 patient days were 8.9 in NICUs and 13.9 in PICUs. Median NICU NI device associated rates by birth weight (>2500 g, 1501-2500 g, 1001-1500 g, and 50% reversibility) occurred in 38% of patients with Q waves and in 70% of those without Q waves. There was also good agreement between the site of ST-segment elevation and the site of ischemia by perfusion imaging (79%) and between the site of ST-segment elevation and the location of the vessel with significant coronary stenosis (95%). CONCLUSIONS: Patients with dobutamine-induced ST-segment elevation have a depressed left ventricular ejection fraction, a high frequency of multivessel disease, and markedly abnormal myocardial perfusion tomography. In patients with ST-segment elevation and abnormal Q waves, substantial scarring and superimposed ischemia coexist, whereas in patients without Q waves, ST-segment elevation usually denotes severe ischemia. PMID- 11391308 TI - Optimal 3-dimensional method for right and left ventricular fourier phase analysis in electrocardiography-gated blood-pool SPECT. AB - BACKGROUND: To overcome some of the limitations imposed by planar imaging, we aimed to optimize the use of first harmonic Fourier phase analysis (FPA) in electrocardiography-gated blood-pool single photon emission computed tomography (GBPS) by comparing different quantitative, 3-dimensional methods. METHODS AND RESULTS: Three groups of patients who underwent GBPS were evaluated: group 1, 8 patients with no heart disease; group 2, 10 patients with left ventricular disease; and group 3, 6 patients with right ventricular disease. Six different methods for FPA were compared: surface, cylindrical, spherical, and hybrid methods with fixed thresholding and spherical and hybrid methods with multiple thresholding. The hybrid method with multiple thresholding for the left ventricle and the spherical method for the right ventricle provided the highest discrimination score (phase) between normal and abnormal ventricles. Among methods with similar discrimination score for these 2 methods, the cylindrical and hybrid methods for the left ventricle and the spherical method for the right ventricle provided the best homogeneity of phase distribution histogram in normal ventricles. These were considered the optimal methods for FPA. CONCLUSIONS: The hybrid or cylindrical method for the left ventricle and the spherical method for the right ventricle with fixed thresholding are the optimal methods for FPA in GBPS. PMID- 11391309 TI - Oxidative stress in cardiovascular disease. AB - The oxidative hypothesis of cardiovascular disease (CVD) has undergone tremendous changes during the past few years. Innumerable new proatherogenic effects have been added to the existing list that could be attributed to oxidative stress. However, both animal and human trials with a variety of antioxidants have failed to establish unambiguously a protective role for antioxidants in the prevention of CVD. This could be because of poor choice and dosage of antioxidants, incompatible experimental models, oxidative metabolism of the antioxidant, unrealistic expectations, and other reasons. More importantly, recent studies suggest that oxidative stress also could induce antioxidant enzymes in both cell culture and in vivo systems. In lieu of the potential of known deterrents of CVD such as exercise, estrogens, and the consumption of polyunsaturated fatty acids to induce an oxidative stress, the possibility that natural enhancement of antioxidant defense in the artery could better serve to deter CVD cannot be ignored. Thus, if oxidative stress does play a role in CVD, it may be better, in the absence of suitable animal models that respond to antioxidants, to adhere to behavioral and dietary changes that have been shown to benefit CVD. PMID- 11391311 TI - Clinical considerations in cardiac stress testing. PMID- 11391313 TI - F-18 fluorodeoxyglucose imaging in myocardial ischemia: beyond myocardial viability. PMID- 11391312 TI - What is wrong with the treadmill exercise test? PMID- 11391310 TI - Cardiac receptor physiology and its application to clinical imaging: present and future. AB - Both gamma imaging and positron emission tomography (PET) imaging of cell surface receptors have become possible through the development of agonists and antagonists with high specific radioactivity and high specificity for the receptors. An understanding of the physiology of the cardiac receptor system is essential to comprehending receptor imaging. The complexity of the physiologic information developed over the past decade has been compounded by the concomitant discovery of additional receptor subtypes. The following is a review of a select group of cardiac receptors and their regulation-namely, adrenergic, muscarinic cholinergic, adenosine, and angiotensin I and II receptors. The role of imaging regional receptor localization and function in providing new insights into cardiac pathology and therapeutic avenues is explored. PMID- 11391314 TI - Detection of a large substernal goiter during Tc-99m tetrofosmin cardiac SPECT imaging. PMID- 11391315 TI - Will current training programs prepare pediatricians to meet the health care needs of children in the 21st century? An opinion. PMID- 11391316 TI - Cerebral palsy in term infants--birth or before birth? PMID- 11391317 TI - Sleep, sleep position, and the sudden infant death syndrome: To sleep or not to sleep? That is the question. PMID- 11391318 TI - Nasal steroids as treatment for obstructive sleep apnea: Don't throw away the scalpel yet. PMID- 11391319 TI - The association of Apgar score with subsequent death and cerebral palsy: A population-based study in term infants. AB - OBJECTIVE: To estimate the risk of adverse outcomes for newborns with a low Apgar score. STUDY DESIGN: Population-based cohort study. All 235,165 children born between 1983 and 1987 in Norway with a birth weight of at least 2500 g and no registered birth defects were followed up from birth to age 8 to 12 years by linkage of 3 national registries. Outcomes were death and cerebral palsy (CP). RESULTS: Five-minute Apgar scores of 0 to 3 were recorded for 0.1%, and scores of 4 to 6 were recorded for 0.6% of the children. Compared with children who had 5 minute Apgar scores of 7 to 10, children who had scores of 0 to 3 had a 386-fold increased risk for neonatal death (95% CI: 270-552) and an 81-fold (48-138) increased risk for CP. If Apgar scores at both 1 and 5 minutes were 0 to 3, the risks for neonatal death and CP were increased 642-fold (442-934) and 145-fold (85-248), respectively, compared with scores of 7 to 10. CONCLUSION: The strong association of low Apgar scores with death and CP in this population with a low occurrence of low scores shows that the Apgar score remains important for the early identification of infants at increased risk for serious and fatal conditions. PMID- 11391320 TI - Congenital abnormalities among children with cerebral palsy: More evidence for prenatal antecedents. AB - OBJECTIVES: To investigate the association between cerebral palsy (CP) and congenital abnormalities among children with very low, low, and normal birth weight. STUDY DESIGN: A population-based, case-control study among the cohort of 155,636 live births delivered between 1983 and 1985 in 4 California counties. Children with moderate or severe congenital CP (n = 192) diagnosed by age 3 were identified from 2 California State service agencies, and 551 control children were randomly sampled from birth certificate files. Information on congenital abnormalities diagnosed by the age of 1 year was obtained from the California Birth Defects Monitoring Program registry. Odds ratios (OR) and 95% CIs were calculated to estimate risk for CP associated with congenital abnormalities. RESULTS: Among singletons, congenital abnormalities were present in 33 (19.2%) children with CP and 21 (4.3%) control children (OR = 5.2, 95% CI 2.8-9.7). For each birth weight group, the percent of children with congenital abnormalities among children with CP exceeded that among control children. Structural abnormalities of the central nervous system were more common among children with CP (OR = 16.2, 95% CI 5.8-49.3) than control children. In contrast, the percent of children with non-central nervous system abnormalities only was similar between case patients and control subjects. CONCLUSION: These findings provide further evidence that factors operating in the prenatal period contribute significantly to the etiology of CP. PMID- 11391321 TI - The prone sleeping position impairs arousability in term infants. AB - OBJECTIVE: To investigate whether the prone sleeping position impaired arousal from sleep in healthy infants and whether this impairment was related to cardiorespiratory variables, temperature, or age. STUDY DESIGN: Healthy term infants (n = 24) were studied with daytime polysomnography on 3 occasions: 2 to 3 weeks after birth, 2 to 3 months after birth, and 5 to 6 months after birth. Multiple measurements of arousal threshold (cm H(2)O) in response to air-jet stimulation applied alternately to the nares were made in both active sleep and quiet sleep when infants slept both prone and supine. RESULTS: Arousal thresholds were significantly higher in both active sleep and quiet sleep when infants slept prone at 2 to 3 weeks and 2 to 3 months, but not at 5 to 6 months. These increases were independent of any sleep position-related change in either rectal or abdominal skin temperature, respiratory rate, oxygen saturation, or heart rate. CONCLUSIONS: The prone position significantly impairs arousal from both active sleep and quiet sleep in healthy term infants. This impairment in arousability occurred with no clinically significant changes in cardiorespiratory variables or body temperature. Decreased arousability from sleep in the prone position provides an important insight into its role as a risk factor for sudden infant death syndrome. PMID- 11391322 TI - Disproportionate alterations in body composition of large for gestational age neonates. AB - OBJECTIVE: The objective was to compare dual-energy x-ray absorptiometry-measured body composition between large (LGA) and appropriate (AGA) birth weight for gestational age neonates. STUDY DESIGN: LGA term infants (n = 47) with birth weights > or =4000 g were compared with 47 gestational age-matched AGA infants; 11 LGA infants were born to mothers with gestational (9) or pregestational diabetes (2). Dual-energy x-ray absorptiometry scans were performed at 1.8 +/- 1.0 days after birth. RESULTS: Body weight and length were the dominant predictors of body composition in LGA and AGA neonates. However, LGA neonates had significantly (P <.001, all comparisons) higher absolute amounts of total body fat, lean body mass, and bone mineral content and had significantly (P <.001, all comparisons) higher proportions of total body fat and bone mineral content but lower lean body mass as a percent of body weight. The changes for total body fat and lean body mass as a percent of body weight were greatest (P <.001) in LGA infants whose mothers had impaired glucose tolerance. CONCLUSION: LGA neonates have higher body fat and lower lean body mass than AGA infants. Impaired maternal glucose tolerance exaggerated these body composition changes. PMID- 11391323 TI - Continuous enteral feeding impairs gallbladder emptying in infants. AB - OBJECTIVE: The objective was to test the hypothesis that continuous enteral feeding impairs gallbladder emptying in infants. STUDY DESIGN: A prospective crossover study was performed in 15 infants: (1) bolus enteral feeds were given in phase A, (2) a continuous milk feed was given for 3 days in phase B, and (3) bolus feeds were resumed in phase C. The gallbladder was studied with ultrasonography in phase A, on days 1 and 3 of phase B, and at the start and on days 2 and 4 of phase C. RESULTS: Baseline volume was 116.1 mm(3) (range, 48.1 to 374.8 mm(3)) in phase A and 293.3 mm(3) (range, 109.4 to 1134.9 mm(3)) (P <.001) after 3 days of phase B; it returned to the phase A value after 4 days of phase C. The contraction index was 65.2% (range, 40.6% to 78.2%) in phase A and 1.7% (range, 0% to 8.4%) (P <.001) after 3 days of phase B. It returned to its phase A value immediately after bolus enteral feeds were resumed in phase C. CONCLUSIONS: Continuous enteral feeding leads to an enlarged, noncontractile gallbladder in infants. Emptying is observed immediately after bolus feeds are resumed, and volume returns to baseline after 4 days. The mode of feeding has important bearings on the motility of the extrahepatic biliary tree. PMID- 11391324 TI - Effect of jaw-thrust and continuous positive airway pressure on tidal breathing in deeply sedated infants. AB - OBJECTIVES: To examine the physiologic impact of the jaw-thrust maneuver or the administration of continuous positive airway pressure (CPAP) on tidal breathing in deeply sedated infants. STUDY DESIGN: Prospective, non-randomized study of infants undergoing elective fiberoptic bronchoscopy while sedated with intermittent doses of propofol. METHODS: Spontaneous tidal breathing was measured in the supine position by means of a spirometer attached to a bronchoscopy face mask. Tidal breaths were recorded under the following conditions: (1) neutral sniffing position, (2) jaw-thrust, (3) neutral sniffing position, and (4) CPAP of 5 cm H(2)O. Improvement was defined as a change of more than twice the coefficient of variation of repeated measurements of tidal volume and flows from baseline. RESULTS: Jaw-thrust increased tidal volume, minute ventilation, and peak tidal inspiratory and expiratory flows significantly in all 13 infants studied (mean +/- SEM age = 8 +/- 2 months). CPAP increased peak tidal inspiratory and expiratory flows by more than twice the coefficient of variation of baseline measurements in 6 patients and tidal volume and minute ventilation in 5 of 10 patients studied. CONCLUSION: Jaw-thrust and CPAP are effective techniques to improve ventilation of sedated infants undergoing interventions that compromise upper airway patency. PMID- 11391325 TI - The association between respiratory viruses and symptoms in 2-week-old infants at high risk for asthma and allergy. AB - OBJECTIVES: The role of viral respiratory tract infections in the onset of childhood asthma and allergy is controversial, partly because of limited understanding about postnatal viral exposures. We investigated the prevalence of 3 common respiratory viruses and associated respiratory symptoms in 2-week-old infants at high risk for having asthma and allergy. STUDY DESIGN: Frozen nasal specimens from 2-week-old children at high risk (n = 495) underwent reverse transcription-polymerase chain reaction (RT-PCR) for picornavirus-, parainfluenza , and respiratory syncytial virus-specific nucleic acid. RT-PCR findings were related to respiratory symptoms (cold, cough, and wheeze) and to characteristics implicated with increased risk for asthma and allergy. RESULTS: Viral RT-PCR was positive in 199 (40.2%) of 495 specimens examined, with picornavirus and parainfluenza significantly associated with respiratory symptoms. Viral prevalence was significantly higher in children born during the winter and summer months. CONCLUSIONS: A high percentage (40.2%) of infants at high risk for asthma and allergy had been exposed to common respiratory viruses at 2 weeks of age. RT PCR is a powerful diagnostic method that can be used in epidemiologic studies examining the role of viral respiratory tract infections in the pathogenesis of pediatric asthma and allergy. PMID- 11391326 TI - Efficacy of fluticasone nasal spray for pediatric obstructive sleep apnea. AB - OBJECTIVE: We tested the hypothesis that a 6-week course of a nasal glucocorticoid spray would decrease the severity of obstructive sleep apnea in children with adenotonsillar hypertrophy. STUDY DESIGN: We conducted a randomized, triple-blind, placebocontrolled, parallel-group trial of nasal fluticasone propionate versus placebo in 25 children aged 1 to 10 years with obstructive sleep apnea proven on polysomnography. The primary outcome was the change from baseline in the frequency of mixed and obstructive apneas and hypopneas. RESULTS: Thirteen children received fluticasone, and 12 received placebo. The mixed/obstructive apnea/hypopnea index decreased from 10.7 +/- 2.6 (SE) to 5.8 +/- 2.2 in the fluticasone group but increased from 10.9 +/- 2.3 to 13.1 +/- 3.6 in the placebo group, P =.04. The mixed/obstructive apnea/hypopnea index decreased in 12 of 13 subjects treated with fluticasone versus 6 of 12 treated with placebo, P =.03. The frequencies of hemoglobin desaturation and respiratory movement/arousals also decreased more in the fluticasone group. Changes from baseline in tonsillar size, adenoidal size, and symptom score were not significantly different between groups. CONCLUSION: Nasal fluticasone decreased the frequency of mixed and obstructive apneas and hypopneas, suggesting that topical corticosteroids may be helpful in ameliorating pediatric obstructive sleep apnea. PMID- 11391327 TI - Long-term comparative trial of positive expiratory pressure versus oscillating positive expiratory pressure (flutter) physiotherapy in the treatment of cystic fibrosis. AB - OBJECTIVE: The objective was to evaluate the long-term effects of physiotherapy with an oscillating positive pressure device ("flutter") compared with physiotherapy with the use of a positive expiratory pressure (PEP) mask in patients with cystic fibrosis (CF). STUDY DESIGN: Forty children with CF were randomly assigned to performing physiotherapy with the PEP mask or the flutter device for 1 year. Clinical status, pulmonary function, and compliance were measured at regular intervals throughout the study. RESULTS: The flutter group demonstrated a greater mean annual rate of decline in forced vital capacity compared with the PEP group (-8.62 +/- 15.5 vs 0.06 +/- 7.9; P =.05) with a similar trend in forced expiratory volume in 1 second (-10.95 +/- 19.96 vs -1.24 +/- 9.9; P =.08). There was a significant decline in Huang scores (P =.05), increased hospitalizations (18 vs 5; P =.03), and antibiotic use in the flutter group. CONCLUSION: Flutter was not as effective in maintaining pulmonary function in this group of patients with CF compared with PEP and was more costly because of the increased number of hospitalizations and antibiotic use. PMID- 11391328 TI - Treatment of vitamin K deficiency in cystic fibrosis: Effectiveness of a daily fat-soluble vitamin combination. AB - OBJECTIVE: Patients with cystic fibrosis (CF) and pancreatic insufficiency (PI) commonly have vitamin K deficiency, and those with CF-associated liver disease (CFLD) have universal vitamin K deficiency. We evaluated the effectiveness of an oral fat-soluble vitamin combination (ADEKs) to treat patients with vitamin K deficiency. STUDY DESIGN: Patients with PI and CF (mean age, 15 years; range, 0.6 to 46 years) including 6 with advanced CFLD were prospectively enrolled in a study of a fat-soluble vitamin combination taken on a daily basis. None had received vitamin K supplementation for at least 4 months before the study. Fat soluble vitamin combination supplementation was given for a minimum of 4 months; the mean vitamin K intake was 0.18 mg/d (SD = 0.1, range, 0 to 0.3). The primary outcome was change in plasma PIVKA-II (prothrombin in vitamin K absence). RESULTS: Before supplementation 58 (81%) of 72 patients had abnormal PIVKA-II levels (>2.9 ng/mL). After supplementation 29 (40%) had abnormal PIVKA-II levels (P =.001). All 6 patients with advanced CFLD had abnormal PIVKA-II levels (median, range of 20.8, 5.5 to 55 ng/mL) before treatment, which corrected to normal in 50% (4.1, 2.1 to 65 ng/mL). Four patients, 2 with CFLD, had a prolonged prothrombin time (>13.5 seconds) at both time periods. CONCLUSIONS: An oral fat soluble vitamin combination with a modest amount of vitamin K can, as a daily supplement, improve the PIVKA-II levels in patients with PI and CF. PMID- 11391329 TI - Bleeding disorders: A common cause of menorrhagia in adolescents. AB - OBJECTIVE: To determine the frequency of underlying bleeding disorders in adolescents with menorrhagia. STUDY DESIGN: We retrospectively reviewed the charts of all girls, aged 10 to 19 years, who presented to our children's hospital for inpatient or outpatient evaluation of menorrhagia between January 1990 and November 1998. RESULTS: At presentation, 9 of the 71 girls (13%) had thrombocytopenia (platelet count <150,000/microL; range, 5000-106,000/microL). The most common causes for thrombocytopenia were immune thrombocytopenic purpura (n = 5) and myelosuppression caused by chemotherapy (n = 2). Of 14 girls who underwent a more detailed hemostatic evaluation, 8 were given a diagnosis of a hereditary coagulation disorder: 6 had platelet function defects and 2 had type 1 von Willebrand disease. Excessive menstrual bleeding commonly results in anemia. One half of the total group had anemia (hemoglobin <12.0 g/dL). Seven girls (10%) had potentially life-threatening anemia (hemoglobin <5.0 g/dL). CONCLUSIONS: Acquired and congenital bleeding disorders are common causes of menorrhagia in adolescent girls. Severe anemia is a frequent complication of menorrhagia. We recommend that adolescents without thrombocytopenia who present with menorrhagia receive a comprehensive hemostatic evaluation, including testing for von Willebrand disease and platelet function defects. PMID- 11391330 TI - Anti-HPA-3A induces severe neonatal alloimmune thrombocytopenia. AB - OBJECTIVE: Fetal and neonatal alloimmune thrombocytopenia (AIT) caused by feto maternal incompatibility at the HPA-1a (PLA-1) locus is well characterized. Alloimmunization and disease caused by HPA-3a is rare. STUDY DESIGN: We conducted a retrospective analysis of all known cases of AIT caused by HPA-3a incompatibility identified at 3 major reference laboratories from 1986 to 1996. Platelet antigen typing and antibody specificity were determined by serologic evaluation. In some cases confirmatory genotyping was performed. RESULTS: Fourteen cases of anti-HPA-3a-induced AIT in 11 families were identified. Five patients had a previous affected sibling, and 2 cases were firstborn children. All patients had severe thrombocytopenia at birth (platelet count <20 x 10(9)/L). Regardless of therapy, the median time to platelet recovery was 6 days (range, 3 to 23 days). Two (15%) patients had documented intracranial hemorrhage, 1 with severe sequelae including apnea and convulsions. A literature review describing 16 additional patients corroborates the finding of severe thrombocytopenia and a significant incidence of intracranial hemorrhage caused by HPA-3a incompatibility. CONCLUSION: AIT caused by incompatibility of HPA-3a is similar in severity to disease caused by incompatibility of HPA-1a. Affected families should be appropriately counseled and considered for antenatal therapy. PMID- 11391331 TI - Antibiotics for the prevention of urinary tract infection in children: A systematic review of randomized controlled trials. AB - OBJECTIVE: The objective was to evaluate the effectiveness of low-dose, long-term antibiotics for the prevention of symptomatic urinary tract infection (UTI) in children. DESIGN: This was a systematic review of randomized controlled trials with a random effects model meta-analysis. PARTICIPANTS: Five trials involving 463 children were performed. RESULTS: Three trials (n = 392) evaluated the effectiveness of long treatment courses of antibiotics (2 to 6 months) for children with acute UTI to prevent subsequent, off-treatment infection. Only 2 trials (n = 71) evaluated the effectiveness of long-term, low-dose antibiotics to prevent on-treatment UTI. Very few of the children enrolled in the trials were boys, had abnormal renal tracts, or were infants. The trial quality was poor, with a lack of blinding, and unstated UTI definitions were almost universal. Long term antibiotic administration reduced the risk of UTI with treatment (relative risk 0.31, 95% confidence limits 0.10 to 1.00), but there was significant heterogeneity (Q = 13.45, P <.01), and there was no sustained benefit once antibiotics had ceased (relative risk 0.79, 0.61 to 1.02). CONCLUSIONS: Methodologic and applicability problems with published trials mean that there is considerable uncertainty about whether long-term, low-dose antibiotic administration prevents UTI in children. Well-designed, randomized, placebo controlled trials are still required to evaluate this commonly used intervention. PMID- 11391332 TI - Unilateral vesicoureteric reflux: Low prevalence of contralateral renal damage. AB - OBJECTIVE: We assessed the risk for the occurrence of renal damage in children with vesicoureteric reflux (VUR). STUDY DESIGN: We reviewed the records of 187 consecutive children, aged 3.8 +/- (SD) 2.8 years, with unilateral primary VUR diagnosed after urinary tract infection (UTI). Dimercaptosuccinic acid renal scintigraphy was performed 4 to 6 months after the last UTI. Three patterns of renal damage were identified: global reduction (GR) of renal radionuclide uptake (20% to 40% of relative uptake), focal defects (FD) in uptake, and shrunken (relative uptake <20%) kidney (SK). We assumed that in these subjects FD indicated postpyelonephritic damage and that GR indicated congenital renal damage. RESULTS: Scintigraphic renal damage of any type was present in 36.9% of the refluxing and in 3.2% of the nonrefluxing kidneys (odds ratio [OR], 17.6; 95% CI, 7.4 to 41.9). FD were present in 15.5% and 2.7% (OR, 6.7; CI, 2.5-17.6), GR in 19% and 0.5% (OR, 44.3; CI, 6.1 to 327.2), and SK in 6.9% and 0%, respectively. Patients with severe VUR showed a higher probability of renal damage than those with nonsevere VUR. CONCLUSIONS: In children with UTI and VUR, the refluxing kidney is most at risk of both congenital and acquired renal damage, and this risk increases with severity of reflux. PMID- 11391333 TI - Long-term treatment with growth hormone in short children with nephropathic cystinosis. AB - OBJECTIVE: The objective was to assess the efficacy and safety of growth hormone (GH) treatment in severely growth retarded children with nephropathic cystinosis during conservative treatment and during renal replacement therapy. STUDY DESIGN: The design was an open-labeled prospective trial with a run-in period of 1 year. RESULTS: A total of 74 children with cystinosis (age 3.0 to 18 years) were treated with GH over a mean period of 3.1 years (range 1 to 10 years); 52 patients were receiving conservative treatment (mean age 7.1 years), 7 were receiving dialysis (12.5 years), and 15 had received a renal transplant (14.8 years). The mean standardized height (SD score) was -4.0 in the conservative treatment group, -4.4 in the dialysis group, and -4.9 in the renal transplant group. During the first treatment year, height velocity doubled in the conservative treatment group, increased by 80% in the dialysis group, and increased by 45% in renal transplant group. Within 3 years the height SD score increased by +1.6 (P <.001) in prepubertal patients receiving conservative treatment, and percentile parallel growth was maintained thereafter. These effects of GH were less expressed in peripubertal patients receiving renal replacement therapy. No major side effects were observed. CONCLUSION: Long-term GH treatment is safe and effective in young children with nephropathic cystinosis. GH treatment should be started early in the course of the disease if adequate nutrition and cysteamine treatment do not prevent growth retardation. PMID- 11391334 TI - Reduced total plasma homocyst(e)ine in children and adolescents with type 1 diabetes. AB - OBJECTIVES: The objective was to investigate total plasma homocyst(e)ine (tHcy), methylenetetrahydrofolate reductase (MTHFR) genotype, and the contribution of diet to homocysteine values in children and adolescents with type 1 diabetes and a control group. STUDY DESIGN: A total of 78 children with type 1 diabetes and 59 members of an age- and sex-matched control group were recruited. Fasting samples were collected for tHcy, MTHFR genotype, serum vitamin B(12), serum folate, red cell folate, and plasma creatinine. Food frequency questionnaires targeted intake of folate, vitamin B(6), and vitamin B(12). RESULTS: Fasting tHcy was reduced in patients compared with the control group (4.7 vs 5.9 micromol/L, P <.001). Serum folate (P =.002), red cell folate(P <.001), and serum vitamin B(12) (P =.005) were higher, and plasma creatinine was lower. A significant difference in tHcy values between patients and the control group persisted after correction was done for these factors (r = 0.1, P =.02). No difference was seen in the frequency of MTHFR polymorphisms. tHcy was not elevated in those patients with the 677TT or 677T/1298C genotypes, although red cell folate was significantly higher in members of the case (P =.01) and control groups (P =.05) with a 677 TT genotype. Dietary intake of folate correlated with serum folate (r = 0.4,P =.005). CONCLUSION: tHcy values are lower in children and adolescents with type 1 diabetes. Higher serum levels of folic acid and vitamin B(12), reflecting differences in dietary intake between children with diabetes and members of a control group, partially account for this difference. PMID- 11391335 TI - The relation between hyperventilation and pediatric syncope. AB - OBJECTIVE: The objective was to evaluate prospectively the role of hyperventilation in the development of neurocardiogenic syncope in children during head-upright tilt testing (HUTT). STUDY DESIGN: Tilt testing was performed in 34 children (mean age 10.6 years) with clinical suspicion of syncope. Respiratory rate and end-tidal carbon dioxide pressure were continuously monitored during HUTT. RESULTS: Tilt test was negative in 12 cases; 3 (25%) patients of this group exhibited hyperventilation any time during the test. In the remaining 22 cases the HUTT was positive, and 15 (68.2%) patients of this group exhibited hyperventilation at the onset of clinical symptoms and during syncope. In the positive HUTT group, the mean time elapsed from the tilt to the onset of syncope and the mean time elapsed from the onset of clinical symptoms to syncope (latency time) were significantly longer for patients who hyperventilated than for those who did not hyperventilate, (21.8 vs 11.5 minutes) (P =.002) and (78 vs 51 seconds) (P =.04), respectively. CONCLUSIONS: Spontaneous hyperventilation could play a relevant role in the pathophysiology of pediatric neurocardiogenic syncope, and it could point out a specific subtype of response to orthostatic stress in susceptible patients. We suggest the inclusion of capnography in tilt-test protocols could improve the assessment of syncope in children. PMID- 11391336 TI - Normal bone mineral density after treatment for childhood acute lymphoblastic leukemia diagnosed between 1991 and 1998. AB - OBJECTIVE: We investigated whether previous reports of reduced bone mineral density after management for childhood acute lymphoblastic leukemia (chALL) were confirmed in a more recently treated cohort. STUDY DESIGN: In a cross-sectional study 75 subjects who were given the diagnosis of chALL between January 1, 1991, and December 31, 1997 (69% standard, 31% high risk), at Denver Children's Hospital and who were 11 to 82 months post-diagnosis with no history of relapse, secondary malignancy, or transplant underwent whole body areal bone mineral densitometry (BMD(A) expressed as age- and sex-standardized z scores), a food frequency questionnaire, and a weight-bearing activity survey. RESULTS: Overall, the mean whole body BMD(A) z score was normal (+0.22 +/- 0.96). A significant positive association was found with whole body BMD(A) z score and years elapsed since the beginning of maintenance (linear regression coefficient = +0.2 Deltaz score/year; 95% CI = 0.09 to 0.3) after adjustment was done for risk status/age category, history of cranial radiation, and total days hospitalized. No association was found with high risk/older age at diagnosis, nutrient intake, chemotherapy dosage, or weight-bearing activity. CONCLUSION: Contrary to previous reports in which cranial radiation and longer hospitalizations were prominent components of therapy, our study suggests that more recently treated patients with chALL do not have persistent abnormalities of bone mineral density after completion of therapy. PMID- 11391337 TI - Maternal postpartum behaviors and mother-infant relationship during the first year of life. AB - OBJECTIVE: To assess the hypothesis that maternal postpartum behaviors toward the newborn may predict the quality of the maternal-infant relationship during the first year. DESIGN: Prospective, non-randomized, longitudinal cohort study of 174 maternal-infant dyads. METHODS: A Postpartum Parenting Behavior Scale (PPBS) was formulated to measure clearly defined observed maternal behaviors toward the infant shortly after birth. The quality of the maternal-infant relationship was assessed at 6 months after birth with the Nursing Child Assessment Satellite Training (NCAST) Feeding Scale and at 12 months after birth with the NCAST Teaching Scale and Ainsworth Strange Situation. RESULTS: The PPBS score correlated significantly with the Feeding Scale score (r =.27, P <.005) and with the Teaching Scale score (r =.23, P <.01). Mothers whose infants were later classified as securely attached in the Ainsworth Strange Situation had higher PPBS scores than mothers of infants classified as insecurely attached (mean +/- SD: 5.18 +/- 1.51 vs 4.63 +/- 1.69, respectively, P <.05). In regression models adjusting for social and demographic factors, the PPBS remained a significant predictor of the Feeding Scale score, the Teaching Scale score, and security of attachment. CONCLUSIONS: Maternal behaviors in the immediate postpartum period may aid in predicting quality of the maternal-infant relationship during the subsequent 12 months, suggesting the potential for early identification of suboptimal parenting. PMID- 11391338 TI - Neonatal tracheal obstruction: A mysterious mass that vanished. PMID- 11391339 TI - Are there neurologic correlates of in utero cocaine exposure at age 6 years? AB - At age 6 years, a cohort of non-asphyxiated children, 52 with gestational cocaine exposure and 63 control subjects, underwent a neurologic examination. Groups did not differ on any aspect of the examination. This finding, while reassuring, does not exclude the possibility of more subtle deficits. PMID- 11391340 TI - A longitudinal study of salivary sialic acid in preterm infants: Comparison of human milk-fed versus formula-fed infants. AB - We found that the saliva of preterm infants fed human milk contains twice the level of sialic acid as that in infants fed commercial formulas. The higher sialic acid level suggests greater viscosity and enhanced protection of the mucosal surfaces in breast-fed infants. Human milk itself is a rich source of sialylated oligosaccharides. PMID- 11391341 TI - Administration of recombinant granulocyte colony-stimulating factor to neonates with septicemia: A meta-analysis. AB - A meta-analysis was used to determine whether administering recombinant granulocyte colony-stimulating factor (rG-CSF) to neonates with bacterial septicemia reduces mortality. Five studies were identified, involving 73 rG-CSF recipients and 82 control subjects. Mortality was lower among the rG-CSF recipients (odds ratio, 0.17; CI, 0.03-0.70; P <.05). However, when the non randomized studies were excluded, the P value was.13. For the subgroups "<2000 g" or "neutropenia," the P value was <.02. Thus the routine use of rG-CSF cannot be recommended for all neonates with sepsis. PMID- 11391342 TI - Primary human herpesvirus 6 infection in liver transplant recipients. AB - We detected primary human herpesvirus 6 (HHV-6) infection in 5 infants who received living related liver transplantation from their HHV-6 seropositive mothers. Primary HHV-6 infection was confirmed by demonstrating the seroconversion of HHV-6 antibodies with an immunofluorescence assay, by the isolation of the virus, or both. Seroconversion of HHV-6 immunoglobulin G antibody was demonstrated in all 5 recipients. HHV-6 was isolated from 3 of the 5 recipients between 2 and 3 weeks after transplantation. Moreover, the virus genome was detected in plasma by polymerase chain reaction in 4 of the 5 recipients during the same period. Although the 5 recipients had pyrexia at the time of primary HHV-6 infection, none of the recipients had a skin rash after defervescence. Clinical symptoms disappeared without specific antiviral treatment in all but 1 of the recipients. PMID- 11391343 TI - Current efficacy of postexposure prophylaxis against measles with immunoglobulin. AB - We assessed results after postexposure prophylaxis against measles with immunoglobulin administered intramuscularly. Children were given 0.33 mL/kg of immunoglobulin from commercially available lots containing various titers of measles antibody within 5 days after exposure. In children receiving immunoglobulin with a titer of < or =16 IU/mL, 57% had clinically evident measles. Immunoglobulin preparations with higher measles antibody titers are required. PMID- 11391344 TI - Hirschsprung disease associated with severe cartilage-hair hypoplasia. AB - Cartilage-hair hypoplasia is a chondrodysplasia with a high incidence of Hirschsprung disease. This study suggests that Hirschsprung disease is associated especially with severe cartilage-hair hypoplasia: the patients with Hirschsprung disease had severe growth failure and a higher incidence of alopecia, infections, malignancies, and childhood anemia than the patients with cartilage-hair hypoplasia who did not have Hirschsprung disease. PMID- 11391345 TI - Hepatic hemangioendothelioma associated with production of humoral thyrotropin like factor. AB - We report on 7 patients referred for treatment of hepatic hemangioendothelioma with increased thyrotropin levels. The serum thyroxine level was decreased in 4 and increased in 2. Immunohistochemistry showed positive staining of tumor, but not of normal liver tissue, for thyrotropin. We propose secretion by the tumor of a thyrotropin-like factor. PMID- 11391346 TI - Diabetic ketoacidosis associated with olanzapine in an adolescent patient. AB - Olanzapine (Zyprexa) is an atypical neuroleptic used in adult and pediatric patients for the management of schizophrenia. Common side effects include increased appetite and weight gain. An uncommon but severe adverse effect is the development of diabetic ketoacidosis, reported until now only in adults. We report a case of acute onset diabetic ketoacidosis presenting in a 16-year-old girl during olanzapine therapy. PMID- 11391347 TI - Lymphocytic interstitial pneumonitis, elevated IgM concentration, and hepatosplenomegaly in ataxia-telangiectasia. AB - An 8-year-old girl developed ataxia-telangiectasia. Western blotting of lysate revealed absence of the ATM protein, and 2 mutations in the ATM gene were found. Subsequently, the patient developed increased respiratory symptoms. Open lung biopsy revealed lymphocytic interstitial pneumonitis, which is not characteristic of ataxia-telangiectasia. There was a therapeutic response to glucocorticosteroid treatment. PMID- 11391348 TI - Juvenile dermatomyositis presenting with anasarca: A possible indicator of severe disease activity. AB - Juvenile dermatomyositis is a rare autoimmune disease characterized by inflammation of the muscle, skin, and other organs. Although localized edema is a common feature of juvenile dermatomyositis, generalized edema has been reported infrequently. We describe a patient with juvenile dermatomyositis presenting with anasarca and note that generalized edema has been associated with severe disease activity. PMID- 11391349 TI - Hepatic storage of glycogen in Niemann-Pick disease type B. AB - We report 2 patients with confirmed Niemann-Pick disease, type B, with previous diagnoses of glycogen storage disease based on excessive glycogen on liver biopsy specimens. These cases emphasize the importance of a complete evaluation, including biochemical confirmation, for patients with suspected metabolic storage diseases. PMID- 11391350 TI - Nodular Leydig cell hyperplasia in a boy with familial male-limited precocious puberty. AB - In boys with familial male-limited precocious puberty, an activating mutation of the luteinizing hormone receptor causes Leydig cell hyperplasia, resulting in excess testosterone production. There are no reports of Leydig cell masses in boys with familial male-limited precocious puberty. We describe a 10-year-old boy with familial male-limited precocious puberty who developed Leydig cell nodules. PMID- 11391351 TI - Spiral computed tomography and three-dimensional image reconstruction of an aberrant innominate artery. PMID- 11391352 TI - Macrophage activation syndrome, the first sign of acute lymphoblastic leukemia in children? PMID- 11391353 TI - Religious/cultural causes of vitamin D deficiency in infants. PMID- 11391355 TI - Parental wishes and poor outcomes in preterm infants. PMID- 11391357 TI - Rickets in Nigerian children. PMID- 11391359 TI - Duplicate publication/duplicate submission-unfortunately still a problem. PMID- 11391360 TI - Redundant surgical publications: tip of the iceberg? AB - BACKGROUND: A redundant publication is one which duplicates previous, simultaneous, or future publications by the same author or group or, alternatively, could have been combined with the latter into one paper. As there is no information about the extent of this problem in the surgical literature, we set out to assess the incidence, spectrum, and salient characteristics of redundant publications in 3 leading surgical journals. METHODS: Original articles (excluding reviews, editorials, abstracts, and letters) published during 1998 in the journals Surgery, The British Journal of Surgery, and Archives of Surgery were searched by using the on-line search engine PUBMED. Each original article was scrutinized to identify redundancy by combining the names of the first, second, and last authors with a few key words from the title. Papers were defined as "suspected" redundant publications if they were found to address the same topic as the "index" article and shared some or most of the elements of methodology, results, or conclusions. The full versions of all suspected papers were retrieved and compared with the index articles. A grading system was developed to define several types of redundant publications: A. "dual"; B. "potentially dual"; C. "salami-slicing." RESULTS: A total of 660 articles were screened. There were 92 index articles (14%) leading to 147 suspected papers found in other journals, representing some potential form of a redundant publication. The vast majority of suspected papers were published within approximately a year of the index paper and were not cited by the latter. Most (69%) of the suspected papers were also published in surgical journals. Only 12 (8.1%) appeared in, or originated from, a "local-foreign" journal. Twenty (13.6%) of the suspected papers met the criteria for dual publications, 50 (34%) for potentially dual publications, and 77 (52.4%) were considered products of salami slicing. CONCLUSIONS: Almost 1 in every 6 original articles published in leading surgical journals represents some form of redundancy. Current on-line search technology provides an effective tool for identifying and tracing such publications, but it is not used routinely as part of the peer review process. Redundancies occur in several well-defined patterns; the phenomenon is widespread, and it cuts across the entire spectrum of surgeons in the United States and abroad. Redundant publications must be recognized not as a mere nuisance but as a real threat to the quality and intellectual impact of surgical publishing. PMID- 11391361 TI - Consensus statement on submission and publication of manuscripts. PMID- 11391362 TI - Clinical applications of ischemic preconditioning: from head to toe. PMID- 11391363 TI - Invited commentary: is there a distinctively surgical ethics? PMID- 11391364 TI - Epidural anesthesia-analgesia shortens length of stay after laparoscopic segmental colectomy for benign pathology. AB - BACKGROUND: Aggressive postoperative care plans after open colectomy may allow earlier discharge, especially in conjunction with preoperative thoracic epidural anesthesia-analgesia using a local anesthetic and narcotic. The purpose of this study was to evaluate the role of thoracic epidural anesthesia-analgesia using bupivacaine and fentanyl citrate in reducing lengths of stay after laparoscopic colectomy (LAC). METHODS: A consecutive cohort of patients who underwent LAC and who received perioperative thoracic epidural anesthesia-analgesia (TEG) was compared with a standard group of patients (STD) undergoing LAC during the 2 months preceding the implementation of the epidural management protocol. Patients with TEG received 6 to 8 mL bupivacaine (0.25%) and fentanyl citrate (100 microg) through a T8-9 or a T9-10 epidural catheter before the incision was made and a postoperative infusion of bupivacaine (0.1%) and fentanyl citrate (5 microg/mL) at 4 to 6 mL/h for 18 hours. STD patients had supplemental intravenous morphine. The postoperative care plan was otherwise identical between the 2 groups. Patients were matched by sex, age, and type of segmental resection. Discharge criteria included tolerance of 3 general diet meals, passage of flatus or stool, and adequate oral analgesia. Length of stay was defined as the time from admission for the surgical procedure to discharge from the hospital. Statistical analysis included a Student t test, Wilcoxon rank sum test, chi-square trend test, and Fisher exact test where appropriate. Data are presented as mean +/- SEM. RESULTS: Procedures performed were: right hemicolectomy-ileocolectomy (TEG, n = 5; STD, n = 5); or sigmoid colectomy-rectopexy (TEG, n = 17; STD, n = 17). There was no significant difference with respect to operating room (OR) time (TEG, 102 +/- 12 minutes; STD, 87 +/- 17 minutes), body mass index (TEG, 26 +/- 2; STD, 26 +/- 2), or American Society of Anesthesiologists class (I-III) distribution (TEG, 3/12/10; STD, 4/11/7), or mean incision length (TEG, 3.5 +/- 0.4 cm; STD, 3.7 +/- 0.3 cm.) No postoperative complications or readmissions occurred in either group. The length of stay decreased in the TEG group (TEG, 2.8 +/- 0.2 days; STD, 3.9 +/- 0.3; P <.001) and the median length of stay for the 2 groups was similarly less (TEG, 2 days; STD, 3 days). CONCLUSIONS: These data suggest that thoracic epidural anesthesia-analgesia has a significant and favorable impact on dietary tolerance and length of stay after LAC. A thoracic epidural appears to be an important component of a postoperative care protocol, which adds further advantage to LAC without the need for labor-intensive and costly patient care plans. PMID- 11391365 TI - Prognostic significance of lymph node involvement in middle and distal bile duct cancer. AB - BACKGROUND: Although lymph node involvement is considered an important prognostic factor, a detailed analysis has not been conducted in middle (Bm) and distal (Bi) bile duct cancer. METHODS: The histopathology of resections taken from 59 patients with Bm and Bi disease (Bm, 33 patients; Bi, 26 patients) was examined. The prevalence of lymph node involvement and its relationship to recurrence and prognosis were investigated. Survival rates were investigated according to the number of metastatic lymph nodes found, the TNM nodal stages, and the nodal stage classifications of The General Rules of the Japanese Society of Biliary Surgery. RESULTS: The frequency of nodal involvement in Bm and Bi was 45.5% and 30.8%, respectively. A significant correlation existed between a patient's prognosis and his TNM nodal stage, Japanese Society of Biliary Surgery nodal stage, and the number of metastatic lymph nodes found (P <.0001, respectively). Among 8 sites of postoperative recurrence, metastasis occurred most frequently in the liver (16/23). Patients with nodal involvement had a significantly higher rate of liver metastasis (10/23) than those without it (6/36) (P =.024). CONCLUSIONS: The number of metastatic lymph nodes found in patients with Bm or Bi cancer, and the nodal stage of their nodes, are significant prognostic indicators. Patients with nodal involvement are at high risk for liver metastasis in Bm and Bi disease. PMID- 11391366 TI - Skip metastases in colon cancer: assessment by lymph node mapping using molecular detection. AB - BACKGROUND: Colon cancer has been assumed to spread sequentially through the regional lymphatic bed, with skip metastases occurring in only 1% to 3% of cases. Molecular techniques allow the detection of occult metastases, but to date have not been applied to assess the pattern of regional lymphatic spread of colon cancer. METHODS: Fifty-five tumors from 54 patients with colonic adenocarcinoma were studied. Lymph node mapping was performed on fresh colonic specimens recording the position of each node on an anatomical diagram. Half of each lymph node was submitted for routine histology examination and half assayed for keratin 20 gene expression by reverse transcription-polymerase chain reaction. Logistic regression was used to analyze the distribution of histologic and occult metastases. RESULTS: A total of 1084 lymph nodes were dissected (median, 19 nodes; range, 4-52). Sixty-four lymph nodes from 20 tumors had histologically evident metastases and 76 lymph nodes from 13 tumors had occult metastases. There was no difference in the distribution of either histologic or occult metastases among paracolic, intermediate, and apical node groups. Ten patients had evidence of anatomical skip lesions after lymph node mapping and molecular analysis, only 1 of which was histologically detectable. CONCLUSIONS: This study demonstrates a higher incidence of skip metastases in colon cancer assessed by molecular techniques than has previously been reported, challenging the concept of sequential development of early lymph node metastases. PMID- 11391367 TI - Hepatocellular carcinoma with biliary tumor thrombi: aggressive operative approach after appropriate preoperative management. AB - BACKGROUND: The aim of this study was to clarify clinicopathologic characteristics of, and to evaluate an aggressive treatment strategy for, hepatocellular carcinoma with biliary tumor thrombi. METHODS: From 1980 to 1999, a total of 132 patients underwent hepatectomy for hepatocellular carcinoma. Of these, 17 patients had macroscopic biliary tumor thrombi and were retrospectively analyzed. RESULTS: The operative procedures included right hepatic trisegmentectomy (n = 1), right or left hepatic lobectomy (n = 11), and segmentectomy or subsegmentectomy (n = 5). In 13 patients, tumor thrombi extended beyond the hepatic confluence and was treated by thrombectomy through a choledochotomy in 8 patients and extrahepatic bile duct resection and reconstruction in 5 patients. The 3- and 5-year survival rates were 47% and 28%, respectively, with a median survival time of 2.3 years. These survival rates were similar to those achieved in 115 patients without biliary tumor thrombi. In a multivariate analysis, expansive growth type and solitary tumors were independent prognostic variables for favorable outcome after operation, whereas biliary tumor thrombi was not a significant prognostic factor. CONCLUSIONS: Surgery after appropriate preoperative management of hepatocellular carcinoma with biliary tumor thrombi yields results similar to those of patients without biliary involvement. Hepatectomy with thrombectomy through a choledochotomy appears to be as effective as a resection procedure. PMID- 11391368 TI - Carcinoma in the porcelain gallbladder: a relationship revisited. AB - BACKGROUND: Gallbladder cancer is the most common biliary tract malignancy. Calcification of the gallbladder wall is reported to be associated with gallbladder cancer. In the literature, the incidence is quoted to be between 12% and 61%. This study aims to clarify the risk of cancer in a calcified gallbladder. METHODS: The charts and pathology reports at the Massachusetts General Hospital were reviewed, and patients with either gallbladder cancer or a calcified gallbladder were included in the study. The Fisher exact test was used to test for the association between cancer and gallbladder wall calcifications. RESULTS: From 1962 to 1999, there were approximately 25,900 gallbladder specimens analyzed at the Massachusetts General Hospital. There were 150 patients with gallbladder cancer and 44 patients with calcified gallbladders. Two types of calcified gallbladders were noted: those with complete intramural calcification (n = 17) and those with selective mucosal calcification (n = 27). The incidence of cancer arising in a gallbladder with selective mucosal wall calcification was approximately 7%. There was a significant association between gallbladder cancer and selective mucosal calcification with an odds ratio of 13.89 (P =.01). There were no patients with diffuse intramural calcification and cancer. CONCLUSIONS: A calcified gallbladder is associated with an increased risk of gallbladder cancer, but at a much lower rate than previously estimated. The incidence of cancer depends on the pattern of calcification; selective mucosal calcification poses a significant risk of cancer whereas diffuse intramural calcification does not. PMID- 11391369 TI - Clinical significance of lymph node micrometastasis in gallbladder carcinoma. AB - BACKGROUND: This retrospective study was intended to define the clinical significance of lymph node micrometastasis in gallbladder carcinoma. METHODS: A total of 1136 regional lymph nodes taken from 63 consecutive patients undergoing radical resection were examined histologically. Micrometastasis was defined as a metastasis missed on routine histologic examination with hematoxylin-and-eosin but detected by immunohistochemical examination with an antibody against cytokeratins 8 and 18. RESULTS: None of 9 patients (0%) with pT1 disease and 19 of 54 patients (35%) with pT2-4 disease had nodal micrometastases. Univariate analysis identified nodal micrometastasis, type of radical resection, M classification, pT classification, perineural invasion, pTNM stage, timing of radical resection, lymphatic vessel invasion, and pN classification as significant variables. Multivariate analysis revealed that nodal micrometastasis (P =.0003) and type of radical resection (P=.0044) were independent prognostic factors. Nodal micrometastasis affected survival adversely, despite the absence (P=.0002) or presence (P <.0001) of overt nodal metastasis. Nodal micrometastasis correlated significantly with invasive characteristics: lymphatic vessel invasion, perineural invasion, and distant metastasis. CONCLUSIONS: Lymph node micrometastasis is the strongest independent predictor of worse survival regardless of the overt nodal status and may indicate aggressive tumor biology among patients undergoing curative resection for gallbladder carcinoma. PMID- 11391370 TI - Characterization of early gastric cancer and proposal of the optimal therapeutic strategy. AB - BACKGROUND AND AIMS: The optimal protocol of the treatment for early gastric cancer has not been fully established. The current study was designed to elucidate the relationship between the depth of tumors with or without an ulcer and the presence of lymph node metastasis and to establish the optimal and practical therapeutic strategy for patients with early gastric cancer. PATIENTS AND METHODS: A retrospective analysis of 1051 patients with early gastric cancer treated by gastrectomy with D1 or D2 lymph node dissection was performed. The patients were divided into those with mucosal (M) tumors and those with submucosal (SM) tumors. These 2 groups were subclassified, depending on the coexistence of ulcer or the degree of submucosal invasion, and were characterized in relation to clinicopathologic factors and 5-year prognosis. RESULTS: The incidence of lymph node metastases from SM tumors (19.8%, 85 of 430) was more frequent than that from M tumors (2.3%, 14 of 621) (P <.001). All M tumors with lymph node involvement, including tumors smaller than 1.5 cm in diameter, had ulceration or ulceration scar in the lesions. SM tumors that had invaded less than 200 microm in depth (SM1a) had significantly less lymph node involvement than those with deeper invasion. The node metastases were confined to epigastric lymph nodes (N1) in both M tumors with ulceration or ulceration scar and SM1a tumors. CONCLUSIONS: All macroscopic M tumors without ulceration or ulceration scar should be considered for endoscopic mucosal resection. The need for reoperation for a formal gastrectomy with lymphadenectomy or a limited surgical operation will vary depending on the pathologic analysis of endoscopic mucosal resection specimens (depth of invasion, presence of ulceration). PMID- 11391371 TI - Can localization studies be used to direct focused parathyroid operations? AB - BACKGROUND: There is considerable controversy today concerning the most appropriate surgical approach for patients with primary hyperparathyroidism. The conventional surgical operation involves a bilateral neck exploration through a collar incision with identification of all parathyroid tissue and removal of abnormal parathyroid glands while the patient is under general anesthesia. The success rate of this operation is about 95% or greater in the hands of an experienced endocrine surgeon. Preoperative localization techniques are generally considered to be unnecessary before initial parathyroid operations. The purpose of this investigation was (1) to evaluate the individual and combined accuracy of ultrasonography and technetium 99m sestamibi scans in localizing abnormal parathyroid glands and (2) to determine whether such scans could be used to direct a focused operation. METHODS: We retrospectively studied 338 patients with sporadic primary hyperparathyroidism who had preoperative neck localization studies, ultrasonography and/or technetium 99m sestamibi scans, and parathyroid exploration (238 patients or, reexploration, 60 patients) from January 1996 to April 2000 at the University of California San Francisco/Mount Zion Medical Center. The preoperative localization studies were recorded as true-positive, false-positive, and false-negative and compared with the surgical and pathologic findings and with the outcome of the operation. RESULTS: All of the abnormal parathyroid glands were correctly identified by ultrasonography in 184 of 303 patients (60.7%) and by technetium 99m sestamibi scanning in 183 of 237 patients (77.2%). The sensitivities of ultrasonography and sestamibi were 65% and 80%, respectively. Among the 202 patients who received both ultrasonography and sestamibi scans, a parathyroid tumor was identified at the same site in 105 (52%) of them. When both techniques identified a parathyroid tumor at the same site, the tests were correct in 101 of 105 patients and the sensitivity increased to 96%. CONCLUSIONS: When both the ultrasonography and sestamibi scans identified the same, solitary parathyroid tumor in patients with sporadic primary hyperparathyroidism, this was the only abnormal parathyroid gland in 96% of the patients. A focused parathyroidectomy could therefore be performed in such patients with an acceptable ( approximately 95%) success rate. PMID- 11391372 TI - Plasma amylin concentration is related to the severity of intestinal ischemic injury in rats. AB - BACKGROUND: Previous work has demonstrated that intestinal ischemia increases plasma amylin concentration. This study examined the relationship between the degree of intestinal ischemia injury and plasma amylin in an experimental rat model. METHODS: Wistar rats were divided into a control group (n = 6); a sham operated group (n = 9); and 3 intestinal ischemia-reperfusion groups (n = 8 in each), which underwent clamping of the superior mesenteric artery for either 15, 30, or 45 minutes followed by 15 minutes of reperfusion. Samples were then collected for intestinal histology and measurement of amylin, insulin, and glucose. RESULTS: There was a positive correlation between the histologic score of the intestinal injury and the measured plasma amylin concentration (R = 0.48, P =.007). The median plasma concentration of amylin was 62 pmol/L (range, 42-97 pmol/L) in the 30-minute intestinal ischemia group and 58 pmol/L (42-86 pmol/L) in the 45-minute intestinal ischemia group. Both these groups were increased compared with the sham-operated group (29 pmol/L; range, 22-57 pmol/L; P <.001 and P <.005, respectively) and the control group (28 pmol/L; range, 26-42 pmol/L; P <.001 and P <.0005, respectively). The median plasma concentration of insulin in the 30-minute intestinal ischemia group was 4230 pmol/L (range, 1360-5770 pmol/L), which was increased compared with both the control group (950 pmol/L; range, 550-1510 pmol/L; P <.005) and the sham-operated group (720 pmol/L; range, 280-4180 pmol/L; P<.005). There were no differences between any of the other groups either for glucose, insulin, or amylin. CONCLUSIONS: Plasma amylin concentration is related to the severity of intestinal ischemic injury. PMID- 11391373 TI - Linkage of oxidative and nonoxidative ethanol metabolism in the pancreas and toxicity of nonoxidative ethanol metabolites for pancreatic acinar cells. AB - BACKGROUND: Alcohol abuse is a major cause of pancreatic damage. Recent experimental evidence suggests that fatty acid ethyl esters (FAEE), nonoxidative ethanol metabolites, injure pancreatic acinar cells. Linkage between oxidative and nonoxidative metabolism of ethanol in the pancreas may contribute to increased FAEE levels. METHODS: To study the association between oxidative and nonoxidative ethanol metabolism, FAEE concentration and FAEE synthase activity in rat pancreatic and liver homogenates incubated with ethanol were evaluated with and without inhibitors of oxidative ethanol metabolism. For toxicity studies, trypsinogen activation peptide synthesis as a measure of pancreatic cell injury was quantitated in unstimulated and cerulein-stimulated isolated pancreatic acinar cells incubated with ethanol or FAEE. RESULTS: Inhibition of oxidative ethanol metabolism results in a 2- to 3-fold increase in nonoxidative ethanol metabolism to FAEE in pancreas and in liver. Both ethanol and FAEE induce increased intracellular trypsinogen activation by more than 50% in the presence of physiologic concentrations of cerulein in vitro. CONCLUSIONS: These findings demonstrate that the inhibition of oxidative ethanol metabolism results in an increase in flux through the nonoxidative pathway and support the proposition that alcohol-induced pancreatic injury is mediated at least in part by FAEE, which are important products of pancreatic ethanol metabolism. PMID- 11391374 TI - Real-time assessment of intraperitoneal tumor growth in a rat model using CEA immunoscintigraphy. AB - BACKGROUND: In most preclinical models, assessment of intraperitoneal tumor location and size require killing the animal. The dynamics of postoperative intraperitoneal tumor implantation and growth remain unclear. A noninvasive method allowing reliable in vivo, real-time assessment of tumor growth is desirable. METHODS: An intraperitoneal tumor homograft using cultured CC531 colorectal cells was created by laparotomy in 24 Wistar Albino Glaxo rats. Eight additional rats were used as controls. Then, 10 MBq technetium 99m-labeled anticarcinoembryonic antigen (anti-CEA) monoclonal antibodies were administrated intravenously and radioactivity uptake was measured by using extracorporeal gamma counting at various time points. Subsequently, the animals were killed for tumor weighting. In 2 more groups of 8 animals, real-time, repeated measures were performed. RESULTS: Correlation between gamma counting and tumor weight was highly significant (P <.001). The regression equation obtained by using the least squares method was: tumor weight (g) = 2.422 + 0.267 x counts. It was possible to obtain real-time tumor growth curves when repeated measurements of radioactivity were performed. At day 25, the predicted tumor weight was 8.49 +/- 0.76 g, the measured weight was 8.16 +/- 0.99 g. CONCLUSIONS: Immunoscintigraphic measurements with technetium 99m anti-CEA antibodies are highly correlated with tumor weight in this model. As opposed to other tumor graft models based on autopsy findings, real-time monitoring is possible. This will allow dynamic studies of intraperitoneal tumor implantation and growth and will reduce the number of animals used in further studies. PMID- 11391375 TI - Loss of T-cell receptor-CD3zeta and T-cell function in tumor-infiltrating lymphocytes but not in tumor-associated lymphocytes in ovarian carcinoma. AB - BACKGROUND: Impaired T-cell function has been noted in tumor-infiltrating lymphocytes (TIL). Recently, loss of function was found to be associated with modifications in T-cell receptor complex (TCR)-mediated signaling. A common feature is loss or reduced expression levels of the signaling chain, TCRzeta. We evaluated whether loss of function in TIL and tumor-associated lymphocytes (TAL) from patients with ovarian cancer is associated with changes in TCRzeta expression, and which factors can cause these defects. METHODS: TIL and TAL were isolated from multiple patients and evaluated for their proliferative capacity by stimulation with a polyclonal stimulus. In addition, expression of TCRzeta and CD3epsilon was evaluated in fresh TIL and TAL by the Western blot technique. Finally, various conditions within a tumor environment were tested for their effect on TCRzeta and CD3epsilon. RESULTS: TIL, but not TAL, were significantly impaired in their proliferative response, even when both populations were derived from the same patient (P <.05). Reduced proliferation levels were associated with loss of expression of TCRzeta but not of CD3epsilon. Exposure of normal T cells to relative ischemia or heat shock, or culture in medium without IL-2, did not significantly reduce expression of TCRzeta compared with CD3epsilon. However, coculture of T cells with tumor-derived macrophages or tumor-derived factors led to a selective loss of TCRzeta compared with CD3epsilon (P <.05). Further analysis suggested that oxides such as hydrogen peroxide secreted by macrophages may be responsible for loss of TCRzeta and high molecular weight factors secreted by certain tumors. CONCLUSIONS: TIL but not TAL show impaired T-cell function, which is associated with loss of TCRzeta. In addition to macrophages secreting oxides, loss of TCRzeta may be caused by tumor-derived soluble factors. PMID- 11391376 TI - Aggressive preoperative management and extended surgery for inflammatory pseudotumor involving the hepatic hilum in a child. AB - Inflammatory pseudotumors involving the hepatic hilum are rare. Only 14 cases have been reported (Table). Liver transplantation has been required when the pseudotumor has invaded extensively into the right and left lobes. (1,2) However, transplantation is associated with the lifelong use of immunosuppressants. This is particularly problematic in children. we report a case of a 6-year-old boy with an inflammatory pseudotumor extensively invading the hepatic hilum who was treated with aggressive surgical excision using the techniques devised for the treatment of hilar cholangiocarcinoma. (3) PMID- 11391378 TI - Dysphagia aortica: harbinger of aortoesophageal fistula? PMID- 11391379 TI - A leukemoid reaction in a patient with a dedifferentiated liposarcoma. PMID- 11391380 TI - A new technique for insertion of totally implantable venous access devices (TIVAD). PMID- 11391381 TI - Long-term outcome of children with papillary thyroid cancer. PMID- 11391382 TI - Selection bias in meta-analysis on acute appendicitis. PMID- 11391384 TI - Salivary glands as test object for assessment of biological compatibility in dentistry. PMID- 11391385 TI - Early afferent reactions in caudate nucleus and neocortex during consolidation and extinction of conditioned. AB - Experiments in cats revealed a clear-cut dependence of the early complex (up to 100 msec) of evoked potentials recorded in the caudate nucleus and temporal cortex on biological relevance of the stimulus (presence or absence of alimentary signal), with constant physical parameters. The early complex of evoked potentials can reflect activity of the mechanisms preventing non-specific motor response to exteroceptive stimulus, although it does not exclude processing the stimulus-carried information during generation of evoked potentials. PMID- 11391386 TI - Peptidergic correction of the effect of acute hypobaric hypoxia in pregnant rats on progeny. AB - Acute hypobaric hypoxia of pregnant rats led to a significant delay in body weight gain, growth and time of eye opening in newborn rat pups which was paralleled by behavioral changes. Preventive intranasal instillations of peptide mixture (semax and beta-casomorphine-7) to pregnant females prevented the effect of hypoxia on the progeny. PMID- 11391387 TI - Behavioral effects of para-chlorophenylalanine in gonadectomized male rats. AB - Para-chlorophenylalanine restores impaired conditioning and retention of active avoidance reaction in castrated animals and improved their behavior in the open field test. PMID- 11391388 TI - Effect of pollack liver oil on lipid metabolism in rat testes. AB - Rat testes were examined by biochemical and histochemical methods after 1, 3, 6, and 12-month pollack liver fat-supplemented diet (0.1 g/kg). Phospholipid concentration in the testicular tissue increased by 16% and cholesterol concentrations decreased by almost 20% after feeding fish oil for 1-3 months, while after 6-12-month diet further increase in cholesterol level in the testes and degeneration of some gonadal cells were noted. The most pronounced changes were seen in more differentiated elements of the spermatogenic epithelium in convoluted tubules. PMID- 11391389 TI - Activities of apoptotic signal 1-regulating protein kinase and poly-(ADP-ribose) polymerase and internucleosomal DNA fragmentation in rat liver during oxidative stress induced by cobalt chloride. AB - We revealed activation of apoptotic signal 1-regulating protein kinase, inhibition of poly-(ADP-ribose) polymerase, and intensification of internucleosomal DNA fragmentation in rat liver during oxidative stress induced by cobalt chloride. PMID- 11391390 TI - Effect of stationary magnetic field on in vivo oxygen binding by blood. AB - The rates of oxyhemoglobin drop and CO(2) release in dogs with magnetized and nonmagnetized blood were compared. Stationary magnetic field enhances blood oxygen capacity by no less than 24%. A hypothesis was proposed on formation of bioxyhemoglobin molecules under the effect of magnetic field, in which hem binds two O(2)molecules. PMID- 11391391 TI - Intracellular distribution and regulation of calpains in the thyroid gland. AB - We studied the distribution of calpains in various subcellular thyrocyte fractions and evaluated the possibility for direct activation of calpain localized in the plasma membrane by thyrotropin. Direct activation of calpain bound to the plasma membrane did not underlie transduction of the thyrotropin signal to Ca(2+)-dependent proteinases. PMID- 11391392 TI - Autoantibodies to beta-amyloid and neurotransmitters in patients with Alzheimer's disease and senile dementia of the Alzheimer type. AB - The content of autoantibodies to beta-amyloid protein Abeta(1-42), its neurotoxic fragment Abeta(25-35), and neurotransmitters were studied in the blood of patients with presenile Alzheimer's disease and senile dementia of the Alzheimer type. Significant differences in the relative content of autoantibodies to Abeta(1-42)and autoantibodies to biogenic amines were demonstrated. These results can be used for the development of a biochemical method for differential diagnosis of Alzheimer dementias. PMID- 11391393 TI - Intracellular localization of VAMP-1 protein in human neutrophils. AB - We studied the intracellular localization of vesicle-associated membrane protein VAMP-1 in human neutrophils. VAMP-1 was associated with membranes of gelatinase and specific secretory granules rapidly mobilized during exocytosis. VAMP-1 probably acts as a component of the SNARE complex during exocytosis of gelatinase and specific granules in human neutrophils. PMID- 11391394 TI - Efficiency of magnesium-containing preparation polykatan in therapy of purulent wounds. AB - Local treatment with polykatan, a magnesium-containing drug based on bischofite mineral, promoted healing of infected skin wounds. Wound cleansing from bacteria was due to a direct antibacterial effect of the drug. PMID- 11391395 TI - Arginine vasopressin fragment AVP(4-9)facilitates induction of long-term potentiation in the hippocampus. AB - Long-term potentiation of CA1 field potentials was induced by weak tetanic orthodromic stimulation of the Schaffer collateral/commissural fibers in isolated hippocampal slices perfused with a medium containing arginine vasopressin fragment AVP(4-9)in micromolar concentrations. It is hypothesized that AVP(4 9)affects induction of long-term potentiation at the intracellular level. PMID- 11391396 TI - Cytokine secreted by rat macrophages and inhibiting proliferation of mesothelial cells. AB - Rat peritoneal macrophages and human peripheral blood monocytes secrete a protein with a molecular weight of 450 kDa, which specifically inhibits proliferation of cultured rat pleural mesothelial cells, but not fibroblasts and epitheliocytes. Protein secretion does not depend on the activation of macrophages. This cytokine is not a cobalamin-binding protein and has no arginase activity. PMID- 11391397 TI - Effects of new taurine derivatives on primary immune response in rats. AB - The effects of new taurine derivatives TAU-15 and TAU-60 with normal and branched alkyl chains, respectively, in a dose of 25 mg/kg on the primary immune response in rats were studied in rats. Intraperitoneal injections of test compounds for 24 days caused transient inhibition of immune reactions to thymus-dependent antigen, which was related to suppressed production of interleukin-1beta playing a key role in antigen presentation. This effect was probably associated with activation of cortisol secretion. TAU-15 inhibited production of tumor necrosis factor-alpha and, therefore, prevented tissue damages. The immune response was normalized after withdrawal of these preparations. PMID- 11391398 TI - Molecular, genetic, and morphological markers during persistence of RNA containing hepatitis C virus in the body. AB - A comparison of 5'-UTR structures in Siberian isolates of hepatitis C virus showed that the majority of genotyped samples had 1b genotype. A correlation was found between HLA antigens and their stable complexes and clinical manifestations of hepatitis C. We revealed immunogenetic criteria of organism's resistance or susceptibility to hepatitis C virus. The presence of hepatitis C virus RNA in the serum determined by the reverse transcription polymerase chain reaction was accompanied by the appearance of pathomorphological signs characteristic of liver tissue infection, but did not correspond to the severity of fibrosis. PMID- 11391399 TI - Abnormal hemopoiesis in long-term bone marrow culture from tumor necrosis factor deficient mice. AB - We studied hemopoiesis in mice deficient by the tumor necrosis factor gene. The total number of cells in long-term bone marrow cultures from these mice 2-fold surpassed that in wild-type and tumor necrosis factor p55 receptor-deficient animals. Increased cell production was related to the absence of tumor necrosis factor expression by hemopoietic precursors. The total cell production by explanted hemopoietic cells from tumor necrosis factor-deficient mice did not depend on the genotype of irradiated stromal sublayer in long-term cell cultures from wild-type mice and animals deficient by tumor necrosis factor or p55 receptor. These results suggest that tumor necrosis factor, but not its p55 receptor, is involved in transduction of signals regulating production of cultured cells. Tumor necrosis factor probably regulates hemopoiesis in long-term bone marrow cultures by initiating apoptosis of hemopoietic cells or inhibiting cell proliferation. Increased cell production probably attests to the absence of one or both effects. PMID- 11391400 TI - Allotransplantation of pituitary cells to rat testicle. AB - Fifty-three transplantations of pituitary tissue (intact and minced with scissors) under the tunica albuginea testis were carried out. In both series pituitary cells retained viability for up to 6 months. Transplantation of cell suspension was not accompanied by the formation of necrotic zone in the transplant; the adjacent testicular tubules and the entire testicular structure remained unchanged, while transplantation of large fragments of the pituitary tissue impaired of spermatogenesis in the adjacent testicular tubules, although most of them remained unchanged. Hence, cell suspension is preferable for transplantation of pituitary tissue into the testicle. PMID- 11391401 TI - Prostaglandin E(2) release by human and Syrian hamster tumor cells and their sensitivity to cytostatic activity of natural killers. AB - The release of prostaglandin E(2) by human and Syrian hamster tumor cells in response to contact with natural killers and their sensitivity to cytostatic activity of natural killers were studied. A previously reported correlation between prostaglandin E(2) release by Syrian hamster tumor cells and their resistance to cytostatic activity of natural killers was confirmed. This resistance can be cancelled by inhibition of prostaglandin E(2) release with indomethacin. Unlike malignant tumor cells of Syrian hamsters, only few human tumor cells resistant to cytostatic activity of natural killers secrete prostaglandin E(2). The resistance of these cell strains to cytostatic activity of natural killers could be cancelled by indomethacin treatment. Possible role of prostaglandin E(2) release as a mechanism of tumor cell protection from effector cells of this type of natural (congenital) immunity is discussed. PMID- 11391402 TI - Potentiation of antitumor effects of cisplatin by tumor necrosis factor-beta. AB - Recombinant tumor necrosis factor-beta potentiated the inhibitory effect of cisplatin on intramuscularly transplanted GA-1 tumor and liver metastasis in mice. The antitumor effect was related to cytotoxic and cytostatic activities of the test preparations rather than to initiation of apoptosis. PMID- 11391403 TI - Effects of transforming growth factor-beta(1)on proliferation of smooth muscle cells in human aortic intima and human promonocytic leukemia THP-1 cells. AB - We studied the effects of transforming growth factor on proliferation of cultured smooth muscle cells from human aortic intima and proliferation and differentiation of human leukemia THP-1 promonocytes. Transforming growth factor inhibited proliferation of these cells, but stimulated differentiation of THP-1 cells. Therefore, transforming growth factor probably modulates proliferation and differentiation of smooth muscle cells and monocytes/macrophages involved in the pathogenesis of atherosclerotic damages. PMID- 11391404 TI - Dynamics of mononuclear phagocytes in lymph nodes and granulomas in chronic tuberculous inflammation. AB - Quantitative changes in mononuclear phagocytes in lymph nodes and tuberculous granulomas in patients with generalized tuberculosis suggest that accumulation of these cells in granulomas occurs due to their recruiting from bone marrow precursors, rather than from lymph nodes. Antimycobacterial therapy leads to dissociation of granulomas accompanied by accumulation of macrophages in lymph nodes. The number of granulomas decreases, but not their size and relative content of epithelioid cells remain unchanged. With due regard of the microanatomy of granulomas, epithelioid cells are regarded as the main site of mycobacterium persistence and the object of targeted drug delivery. PMID- 11391405 TI - Effects of opiate receptor ligands on DNA synthesis in tracheal epitheliocytes and smooth muscle cells of newborn albino rats. AB - The dermorphin analogue A10 injected to newborn rats from the 2nd to 6th day of life increased the index of labeled nuclei in epitheliocytes and smooth muscle cells and labeling intensity in smooth muscle cells. Dynorphin A(1-13)increased the index of labeled nuclei and labeling intensity in epitheliocytes and labeling intensity in smooth muscle cells. Dalargin increased the labeling intensity in epitheliocytes, but had effect on DNA synthesis in muscle cells. PMID- 11391406 TI - Expression of adenylyl cyclase type IX and calcineurin in synapses of the central nervous system. AB - Distribution of type IX adenylyl cyclase and protein phosphatase calcineurin in the brain and in cultured hippocampal neurons from albino rat was immunohistochemically studied. Both enzymes were detected simultaneously in all synaptic structures of most cerebral neurons except for presynaptic sites, where calcium-inhibited type IX adenylyl cyclase was absent. PMID- 11391407 TI - Antiphospholipid syndrome: ultrastructure of microvascular endotheliocytes in musculocutaneous bioptates during systemic lupus erythematosus. AB - Degeneration and atrophy of the epidermis, disorganization of the connective tissue, focal atrophy of skeletal myocytes, and diffuse vasculopathy are the main pathomorphological signs found in musculocutaneous bioptates during the antiphospholipid syndrome associated with systemic lupus erythematosus. The major morphogenetic disorder is alteration of microvascular endotheliocytes accompanied by the formation of concentric perivascular mononuclear infiltrates. PMID- 11391408 TI - Ultrastructural changes in cardiomyocyte mitochondria during regenerative and plastic insufficiency of the myocardium. AB - Daunomycin-induced regenerative and plastic insufficiency of the myocardium was accompanied by accumulation of cardiomyocytes with unstable mitochondrial membranes containing enlarged mitochondria with lightened matrix and fragmented cristae. Total destabilization of mitochondrial membranes was found in cardiomyocytes with most pronounced ultrastructural signs of impaired protein synthesis. These changes in mitochondria were permanent, which suggested that swelling and destruction of cristae were related to intravital decrease in mitochondrial membrane stability. PMID- 11391409 TI - Ultrastructural changes in the nephron and renal protein cleaving function in suckling rabbits with experimental cholera. AB - Immature nephrons and the presence of a proteolytic system mediating extracellular protein digestion in the epithelium of proximal tubules are characteristic features of renal cortical ultrastructure in intact 10-12-day-old suckling rabbits. Cholera infection is accompanied by intensive cleavage of exogenous protein, which starts in podocytes and is completed in the cytoplasm of the proximal tubule epitheliocytes. Overload to nephron associated (in addition to urine production) with intensive extracellular hydrolysis impairs nephrothelium reactivity and increases its vulnerability to cholera exo- and endotoxins and their mediators. PMID- 11391410 TI - Effect of phosphatidylcholine liposome on regeneration of surgical wound in guinea pig lung. AB - Phosphatidylcholine liposomes in a dose of 25 mg/kg displayed wound-healing properties. They increased the count of alveolar macrophages, lymphocytes, and microvessels in damaged regions and, therefore, attenuated emphysematous changes in the lungs. PMID- 11391411 TI - Blood-testis barrier after transplantation of pituitary cell into dystopic testicle. AB - A total of 87 allotransplantations of the pituitary were carried out: into intact testicle with or without subsequent transposition into the abdominal cavity and on a previously created model of unilateral cryptorchidism. Histological study showed that pituitary cells retained viability for up to 3 months in all three series. The main structures of the blood-testis barrier (membranes of testicular tubules, tunica albuginea testis, vascular walls, and interstitial tissue) were not damaged, which determined advantages of dystopic testicle in pituitary cell allotransplantation. PMID- 11391412 TI - A model of combined gastroduodenitis and hyperlipidemia in rats. AB - We elaborated a model of gastroduodenitis combined with hyperlipidemia in rats. This model is based on alimentary lipid disturbances and morphological changes in the gastroduodenal mucosa caused by exogenous damaging factors. Morphological assay revealed gastroduodenitis; biochemical studies of the blood and liver demonstrated hyperlipidemia. This model holds much promise for evaluating pathogenetic mechanisms of combined gastroduodenal and heart diseases. PMID- 11391413 TI - Transmission of tuberculosis in Havana, Cuba: a molecular epidemiological study by IS6110 restriction fragment length polymorphism typing. AB - The combination of molecular and conventional epidemiological methods has improved the knowledge about the transmission of tuberculosis in urban populations. To examine transmission of tuberculosis in Havana, Cuba, with DNA fingerprinting, we studied 51 out of 92 Mycobacterium tuberculosis strains isolated from tuberculosis patients who resided in Havana and whose infection was culture-confirmed in the period from September 1997 to March 1998. Isolates from 28 patients (55%) had unique IS6110 restriction fragment length polymorphism (RFLP) patterns, while isolates from 23 others (45%) had identical patterns and belonged to 7 clusters. Three clusters consisting of six, five and two cases were each related to small outbreaks that occurred in a closed setting. Three other clustered cases were linked to a large outbreak that occurred in another institution. Younger patients were more correlated to clustering than older ones. The finding that 45% of the isolates had clustered RFLP patterns suggests that recent transmission is a key factor in the tuberculosis cases in Havana. The IS6110 RFLP typing made it possible to define the occurrence of outbreaks in two closed institutions. PMID- 11391414 TI - Dynamics of Haematobia irritans irritans (Diptera: Muscidae) infestation on Nelore cattle in the Pantanal, Brazil. AB - From June 1993 to May 1995, horn fly counts were conducted twice a month on untreated Nelore cattle raised extensively in the Pantanal. Horn fly population showed a bimodal fluctuation and peaks were observed every year after the beginning (November/December) and at the end (May/June) of the rainy season, which coincided with mid-late spring and mid-late fall, respectively. Horn flies were present on cattle throughout the year in at least 64% of the animals. Mean horn fly numbers on animals did not exceed 85 flies/cow during peaks and were under 35 flies/cow in most of the remaining periods. The highest infestations (population peaks) were short and dropped suddenly within two weeks. Less than 15% of the animals in both herds could be considered as "fly-susceptible" - showing consistently higher infestations, or "fly-resistant" - showing consistently lower infestations. PMID- 11391415 TI - Simuliid blackflies (Diptera: Simuliidae) and ceratopogonid midges (Diptera: Ceratopogonidae) as vectors of Mansonella ozzardi (Nematoda: Onchocercidae) in northern Argentina. AB - Mansonella ozzardi, a relatively nonpathogenic filarial parasite of man in Latin America, is transmitted by either ceratopogonid midges or simuliid blackflies. In the only known focus of the disease in north-western Argentina the vectors have never been incriminated. This study investigated the potential vectors of M. ozzardi in this area. The only anthropophilic species of these Diptera families biting man at the time of the investigation were Simulium exiguum, S. dinellii, Culicoides lahillei and C. paraensis. Using experimentally infected flies S. exiguum and both species of Culicoides allowed full development of microfilariae to the infective stage, with C. lahillei being a more competent host than S. exiguum. Based on these data, biting rates and natural infectivity rates it is probable that at the begininning of the wet season C. lahillei is the main vector of M. ozzardi and both C. paraensis and S. exiguum secondary vectors. Additionally, it was found that a single dose of ivermectin was ineffectual in eradicating M. ozzardi from infected individuals in this area. PMID- 11391416 TI - Characterization of Triatoma guasayana biotopes in a hardwood forest of Santiago del Estero, Argentina. AB - Triatoma guasayana is a silvatic triatomine species distributed in Argentina, Bolivia and Paraguay. The study was performed in a secondary forest of Santiago del Estero, Argentina. The abundance of T. guasayana was evaluated by census in the following wild biotopes: quimiles (Opuntia quimilo), chaguares (dry bromeliads), logs and underground burrows. Ten biotopes of each type were dismantled in winter (August) and another 40 in summer (January); all fauna was recorded. The biotopes most infested by T. guasayana were quimiles (65%), followed by chaguares (55%), and logs (25%). Quimiles and chaguares were infested in both seasons, whereas logs were positive only in summer and burrows were never infested. Infestation and abundance were higher in summer than in winter. The biotope structure is a key factor for T. guasayana colonization. The larger number of refuges, the constant presence of blood sources and suitable inner microclimatic conditions offered by quimiles may favour the persistence of T. guasayana colonies. The richness of invertebrate fauna per type of biotope was ranked in the same order as that of T. guasayana, suggesting similar microhabitat requirements for all studied arthropods. PMID- 11391417 TI - Environmental characteristics of the cemeteries of Buenos Aires City (Argentina) and infestation levels of Aedes aegypti (Diptera: Culicidae). AB - Cemeteries with many water-filled containers, flowers, sources of human blood, and shade are favorable urban habitats for the proliferation of Aedes aegypti, a vector of yellow fever and dengue. A total of 22,956 containers was examined in the five cemeteries of the city of Buenos Aires, Argentina. The vector was found in four cemeteries that showed an average infestation level of 5.5% (617 positive out of 11,196 water-filled containers). The four cemeteries positive for Ae. aegypti showed significantly different (p<0.01) infestation levels. Vegetation cover and percentage of infestation were significantly correlated (p<0.01), but neither cemetery area nor number of available containers were significantly related to the proportion of positive vases. Our results suggest that the cemeteries of Buenos Aires represent a gradient of habitat favorableness for this vector species, some of which may act as foci for its proliferation and dispersal. PMID- 11391418 TI - Triatoma infestans in Greater Buenos Aires, Argentina. AB - The Health Administration Agencies of many municipalities in Greater Buenos Aires (GBA) receive frequent reports on triatomines in houses. The aim of this work was to identify and describe the dispersal foci of Triatoma infestans in an urban neighborhood of GBA, and contribute to the knowledge of the epidemiological situation in the region. In June 1998, potentially infested places were entomologically evaluated. T. infestans was only detected in a hen building for egg production, which housed approximately 6,000 birds. A total of 2,930 insects were collected. Density was about 9 triatomines/m(2). The proportions of fifth instar nymphs and adults were significantly higher than those of the other stages (p<0.001). The number of triatomines collected largely exceeded the highest domestic infestation found in one house from rural endemic areas of Argentina. Though triatomines were negative for Trypanosoma cruzi, they could acquire the parasite by coming in contact with infected people living in GBA. Besides, the numerous and widely distributed places housing hens and chickens, would favor the settlement of the vector. Together, both facts may constitute a risk of parasitic vectorial transmission. It is recommended to intensify systematic activities of vector search and case detection in GBA. PMID- 11391419 TI - Cebus apella (Primata: Cebidae) as a new host for Fonsecalges johnjadini (Acari: Psoroptidae, Cebalginae) with a description of anatomopathological aspects. AB - Mites collected from the auditory canal of Cebus apella (capuchin monkey), family Cebidae, were identified as Fonsecalges johnjadini (Psoroptidae, Cebalginae). It is the first record of this parasite from this monkey. This paper emphasizes the importance of clinical and anatomopathological examinations for parasitic diagnosis in wild animals. PMID- 11391420 TI - Simulium cuasiexiguum, a new blackfly species (Diptera: Simuliidae) from the Minacu area in the State of Goias, Central Brazil. AB - During studies of Simuliidae at a suspected new focus of human onchocerciasis in central Brazil a new species of Simulium was found. Full descriptions of the adults and pupae of this species, S. cuasiexiguum, are described here, its affinities to closely related species in the subgenus Notolepria are discussed and its distribution in Brazil recorded. PMID- 11391421 TI - Serodiagnosis of chronic Chagas infection by using EIE-Recombinant-Chagas Biomanguinhos kit. AB - A kit based on an enzyme immunoassay, EIE-Recombinant-Chagas-Biomanguinhos, developed by the Oswaldo Cruz Foundation, was evaluated for the serodiagnosis of chronic Chagas disease. Evaluation was performed with 368 serum samples collected from individuals living in an endemic area for Chagas disease: 131 patients in the chronic phase with confirmed clinical, epidemiological, and serological diagnosis (indirect immunofluorescence, indirect hemagglutination or enzyme linked immunosorbent assay) and 237 nonchagasic seronegative individuals were considered negative control. The EIE-Recombinant-Chagas-Biomanguinhos kit showed high sensitivity, 100% (CI 95%: 96.4-100%) and high specificity, 100% (CI 95%: 98 100%). The data obtained were in full agreement with clinical and conventional serology data. In addition, no cross-reaction was observed with sera from patients with cutaneous (n=14) and visceral (n=3) leishmaniasis. However, when these sera were tested by conventional serological assays for Chagas disease, cross-reactions were detected in 14.3% and 33.3% of the patients with cutaneous and visceral leishmaniasis, respectively. No cross-reactions were observed when sera from nonchagasic seronegative patients bearing other infectious disease (syphilis, n=8; HTLV, n=8; HCV, n=7 and HBV, n=12) were tested. In addition, sera of patients with inconclusive results for Chagas disease by conventional serology showed results in agreement with clinical evaluation, when tested by the kit. These results are relevant and indicate that the referred kit provides a safe immunodiagnosis of Chagas disease and could be used in blood bank screening. PMID- 11391422 TI - Comparison of polymerase chain reaction on fresh tissue samples and fecal drops on filter paper for detection of Trypanosoma cruzi in Rhodnius prolixus. AB - PCR detection of Trypanosoma cruzi in Rhodnius prolixus using fresh tissue or fecal drops on filter paper showed comparable results: 38.7% infection rate using the fresh tissue sample and 37.9% by dried fecal drop. PMID- 11391423 TI - Anti-VP1 and anti-VP2 antibodies detected by immunofluorescence assays in patients with acute human parvovirus B19 infection. AB - Acute human parvovirus B19 infection is followed by an antibody response to the structural proteins of the viral capsid (VP1 and VP2). We used 80 sera collected from 58 erythema infectiosum and 6 transient aplastic crisis patients to test IgM and IgG antibodies against these two proteins in an immunofluorescence assay (IFA) using Sf9 cells infected with recombinant baculovirus expressing either VP1 or VP2 antigen. Although less sensitive than IgM capture enzyme immunoassay using native antigen (MACEIA), we could detect anti-VP1 or anti-VP2 IgM antibodies by IFA in 49 patients with acute infection (76.6%). Detection of IgG anti-VP1 and anti-VP2 by IFA, however, was as sensitive as IgG detection by indirect enzyme immunoassay. By applying IgG avidity IFA to sera of the 15 IgM IFA negative patients we were able to confirm acute infection in further 12 cases by IFA. Overall, acute infection was confirmed by IFA in 61 (95.3%) of the 64 patients. PMID- 11391424 TI - Detection of the acute phase of abdominal angiostrongyliasis with a parasite specific IgG enzyme linked immunosorbent assay. AB - Angiostrongylus costaricensis may cause intestinal lesions of varied severity when it accidentally infects man in Central and South America. First-stage larvae have never been detected in stools. Therefore, a parasite-specific IgG ELISA was evaluated for the determination of the acute phase of infection. The specificity and the sensitivity of the immunoassay was shown to be 76.2% and 91.1%, respectively. Eight serum samples taken from patients with histopathological diagnosis, at different time points (3 to 15 months) after surgical treatment, showed a sharp and early decline in antibody reactivity. The titration of anti-A. costaricensis antibodies has proved to be a useful method for the diagnosis of acute abdominal angiostrongyliasis. PMID- 11391425 TI - TT virus infection in children and adults who visited a general hospital in the south of Brazil for routine procedure. AB - TT virus (TTV) is a newly described nonenveloped human virus, with a circular, negative-stranded DNA genome, that was first identified in the blood of a patient with posttransfusion hepatitis of unknown etiology. PCR primers and conditions used for TTV DNA amplification may greatly influence the level of TTV detection in serum. Three PCR assays, with different regions of the genome as targets, were used to test TTV DNA in 130 sera from children and adults visiting a hospital in the south of Brazil, most of them for routine procedure. Forty-four percent of adult sera and 73% of sera from children aged 0-10 years were TTV positive with at least one PCR assay. However, the three assays were able to detect only 33%, 35%, and 70% of the total positive samples. Our results showed a high prevalence of TTV infection in the south of Brazil, particularly among young children, and confirmed the necessity of performing several PCR assays to assess the true TTV prevalence in a determined population. PMID- 11391426 TI - Typing and susceptibility to penicillin of Neisseria meningitidis isolated from patients in Cuba (1993-1999). AB - The susceptibility to penicillin of 111 Neisseria meningitidis strains was assessed by the agar-dilution procedure and serosubtypes were determined by a whole-cell enzyme-linked immunoassay using monoclonal antibodies reagents. Thirty five isolates showed reduced sensitivity to penicillin (MIC > or = 0.1 mg/l and < or = 1 mg/l) and no resistant strains were detected. The most common phenotype was B:4:P1.15 (77.5%) and a rising trend of non-typeable and non-subtypeable strains was detected. The increase in levels of minimal inhibitory concentrations of meningococci to penicillin gives cause for concern and the increase in non typeable and non-subtypeable isolation demand the use of molecular biology techniques for their typing. PMID- 11391427 TI - Isoenzyme profiles of Trypanosoma cruzi stocks from different areas of Paraguay. AB - Twenty one Trypanosoma cruzi stocks from humans, domiciliary triatomines and one sylvatic animal of different areas of Paraguay were subjected to isoenzyme analysis. Thirteen enzyme systems (15 loci in total) were studied. MN cl2 (clonets 39) and SO34 cl4 (clonets 20) were used as references. Relationships between stocks were depicted by an UPGMA dendrogram constructed using the Jaccard's distances matrix. Among the Paraguayan stocks 14 zymodemes were identified (Par1 to Par14), Par 5 being the most frequent. Polymorphism rate and clonal diversity were 0.73 and 0.93, respectively. Average number of alleles per polymorphic locus was 2.5 (range 2-4). These measurements show a high diversity, which is confirmed by the dendrogram topology. All stocks belong to the same lineage, as MN cl2 reference strain (T. cruzi II). Moreover three distinct subgroups were identified and two of them correspond to Brazilian and Bolivian zymodemes, respectively. The third subgroup, the most common in Paraguay, is related to Tulahuen stock. The large geographical distribution of some zymodemes agrees with the hypothesis of clonality for T. cruzi populations. However sample size was not adequate to detect genetic recombination in any single locality. PMID- 11391428 TI - Genetic variability in Brazilian populations of Biomphalaria straminea complex detected by simple sequence repeat anchored polymerase chain reaction amplification. AB - Biomphalaria glabrata, B. tenagophila and B. straminea are intermediate hosts of Schistosoma mansoni, in Brazil. The latter is of epidemiological importance in the northwest of Brazil and, due to morphological similarities, has been grouped with B. intermedia and B. kuhniana in a complex named B. straminea. In the current work, we have standardized the simple sequence repeat anchored polymerase chain reaction (SSR-PCR) technique, using the primers (CA)8RY and K7, to study the genetic variability of these species. The similarity level was calculated using the Dice coefficient and genetic distance using the Nei and Li coefficient. The trees were obtained by the UPGMA and neighbor-joining methods. We have observed that the most related individuals belong to the same species and locality and that individuals from different localities, but of the same species, present clear heterogeneity. The trees generated using both methods showed similar topologies. The SSR-PCR technique was shown to be very efficient in intrapopulational and intraspecific studies of the B. straminea complex snails. PMID- 11391429 TI - Morphometry of submucous and myenteric esophagic plexus of dogs experimentally reinfected with Trypanosoma cruzi. AB - We carried out a morphometric study of the esophagus of cross-bred dogs experimentally infected or consecutively reinfected with Trypanosoma cruzi 147 and SC-1 strains, in order to verify denervation and/or neuronal hypertrophy in the intramural plexus. The animals were sacrificed in the chronic stage, 38 months after the initial infection. Neither nests of amastigotes, nor myositis or ganglionitis, were observed in all third inferior portions of esophageal rings analyzed. No nerve cell was identified in the submucous of this organ. There was no significant difference (p>0.05) between the number, maximum diameter, perimeter, or area and volume of the nerve cells of the myenteric plexus of infected and/or reinfected dogs and of the non-infected ones. In view of these results we may conclude that the 147 and SC-1 strains have little neurotropism and do not determine denervation and/or hypertrophy in the intramural esophageal plexuses in the animals studied, independent of the reinfections. PMID- 11391430 TI - Isotype specific immune responses in murine experimental toxocariasis. AB - In this work, a murine experimental model of toxocariasis has been developed in BALB/c, C57BL/10 and C3H murine strains orally inoculated with 4,000 Toxocara canis embryonated eggs, in order to investigate the isotype-specific immune responses against excretory-secretory antigens from larvae. T. canis specific IgG+M, IgM, IgG, IgA, IgG1, IgG2a and IgG3 were tested by ELISA. The dynamics of the specific immunoglobulins (IgG+IgM) production showed a contrasting profile regarding the murine strain. Conversely to the results obtained with the IgM isotype, the IgG antibody class showed similar patterns to those obtained with IgG+IgM antibodies, only in the case of the BALB/c strain, being different and much higher than the obtained with IgG+IgM antibodies, when the C3H murine strain was used. The antibodies IgG+IgM tested in BALB/c and C57BL/10 were both of the IgM and IgG isotypes. Conversely, in the C3H strain only IgG specific antibody levels were detected. The IgG1 subclass responses showed a similar profile in the three murine strains studied, with high values in BALB/c, as in the case of the IgG responses. PMID- 11391431 TI - Scanning and transmission electron microscopy of the tegument of Paranaella luquei Kohn, Baptista-Farias & Cohen, 2000 (Microcotylidae, Monogenea), parasite of a Brazilian catfish, Hypostomus regani. AB - The surface topography and ultrastructure of the tegument of Paranaella luquei Kohn, Baptista-Farias & Cohen, 2000, a microcotylid monogenean parasite from the gills of Hypostomus regani (Ihering, 1905) (Loricariidae) was studied by scanning (SEM) and transmission electron microscopy (TEM). By SEM, it was observed that the tegument presents transversal ridges, forming folds in the ventral and dorsal surfaces and microvillous-like tegumental projections in the anterior and median regions of body. These projections were also observed by TEM. The tegument is made up of a syncytium delimited by apical and basal plasma membranes, containing inclusion bodies and mitochondria, connected to the nucleated region by means of cytoplasmatic processes. The tegumental cells present a well developed nucleus and cytoplasm containing inclusion bodies, similar to those found on the external layer, mitochondria, rough endoplasmatic reticulum and free ribossomes. PMID- 11391432 TI - Stomach content analyses of Simulium perflavum Roubaud 1906 (Diptera: Simuliidae) larvae from streams in Central Amazonia, Brazil. AB - Stomach contents of Simulium perflavum Roubaud larvae were analyzed and compared with plankton and periphyton collected in five streams, in Central Amazonia (Manaus and Presidente Figueiredo counties), in Sep./Oct.1996 (dry season) and Feb./Mar. 1997 (rainy season). A total of 1,400 last-instar larvae were dissected; the stomach contents were analyzed using different methods: fresh and after oxidation. A total of 87 taxa (algae, diatoms and rotifers) were found in the stomach contents. In each stream, qualitative samples of plankton and periphyton were collected; these were mounted between slides and cover slips. A total of 94 taxa of plankton and 54 taxa of periphyton were collected. One species of Rotifera was present in the stomach contents, plankton and periphyton. Cluster analysis based on species composition of the organisms present in the stomach contents grouped the streams into two major groups, each belonging to a different drainage area. Correlations based on presence/absence of species of microalgae in the stomach contents, plankton and periphyton indicated significant associations (p<0.05) between stomach contents and plankton and between plankton and periphyton (z test); the Sorensen coefficient and cluster analysis corroborate the same associations. PMID- 11391433 TI - The effect of isolation on the life-history traits of Pseudosuccinea columella (Pulmonata: Lymnaeidae). AB - A population of Pseudosuccinea columella was raised under laboratory conditions and its life tables were determined in isolated and paired snails. Isolated snails were significantly larger in shell size than paired snails from five weeks of age onward. Also, statistically significant differences were found for the number of eggs per mass per individual from week 5 to 9, isolated snails exhibiting the highest values. The intrinsic and finite rates of increase were greater in isolated than in paired snails. Either an inhibition of the reproductive output between individuals or the advantage of selfing may be the cause of the differences in this species, acting as a possible mechanism that increase the fitness of isolated snails. PMID- 11391434 TI - A comparative study of preservation and storage of Haemophilus influenzae. AB - The aim of this study was to compare the efficacy of conservation by freezing the strains of Haemophilus influenzae at -20 degrees C and -70 degrees C. Skim milk supplemented with glucose, yeast extract and glycerol allowed highest viability of H. influenzae both at -20 degrees C and -70 degrees C from the media analyzed. Trypticase soy broth and brain heart infusion broth supplemented with glycerol, allowed excellent recovery. Use of cotton swaps as supporting material, with or without addition of cryoprotective agents, did not modify H. influenzae viability after six months of storage. Concentration of the initial inoculum positively affected viability when stored at -20 degrees C. Initial concentration did not influence survival after storage at -70 degrees C. Thawing at room temperature should not exceed 3 h as to get highest survival percentage. PMID- 11391435 TI - Resistance to starvation of Rhodnius neivai Lent, 1953 (Hemiptera: Reduviidae: Triatominae) under experimental conditions. AB - The period of resistance to starvation and the loss of weight until death of Rhodnius neivai in all stages of development were studied. Work was based on experiments conducted under controlled laboratory conditions. One hundred specimens of each nymphal instar were observed: 50 were fed on chicken and 50 on rabbit. Adult females and males were kept together and fed on each host. All bugs were weighed weekly until death. Laid eggs were collected weekly and observed during five weeks to obtain hatchability. Resistance to starvation was similar with both hosts and increased with the evolutionary stage, excepting the 5th nymphal instar and adults. With both hosts, loss of weight was abrupt in the first week and steady in the following weeks. In adults, on the first weeks after eating, there was little or no mortality, after which mortality increased rapidly with the starving time. Reproductive output was higher in the bugs fed on rabbit. R. neivai is among the least resistant triatomine species. PMID- 11391436 TI - Clonal structure of Trypanosoma cruzi Colombian strain (biodeme Type III): biological, isoenzymic and histopathological analysis of seven isolated clones. AB - The clonal structure of the Colombian strain of Trypanosoma cruzi, biodeme Type III and zymodeme 1, was analyzed in order to characterize its populations and to establish its homogeneity or heterogeneity. Seven isolated clones presented the basic characteristics of Biodeme Type III, with the same patterns of parasitemic curves, tissue tropism to skeletal muscle and myocardium, high pathogenicity with extensive necrotic-inflammatory lesions from the 20th to 30th day of infection. The parental strain and its clones C1, C3, C4 and C6, determined the higher levels of parasitemia, 20 to 30 days of infection, with high mortality rate up to 30 days (79 to 100%); clones C2, C5 and C7 presented lower levels of parasitemia, with low mortality rates (7.6 to 23%). Isoenzymic patterns, characteristic of zymodeme 1, (Z1) were similar for the parental strain and its seven clones. Results point to a phenotypic homogeneity of the clones isolated from the Colombian strain and suggest the predominance of a principal clone, responsible for the biological behavior of the parental strain and clones. PMID- 11391437 TI - [Toxic effects and in vitro inefficacy of deltamethrin on larvae of Rhipicephalus sanguineus from Goiania, Goias, Brazil]. AB - It has been observed that formulations of deltamethrin developed for disinsectization of infested places and control of parasitic arthropods of bovines and equines have frequently been used by breeders of dogs to control ticks in their animals and kennels, but without due consideration of the correct dosage or means of application. With the purpose of evaluating the efficacy of this product on Rhipicephalus sanguineus, bioassays were realized with larvae at 27+/-1 degrees C, UR>80% and light period of 12 hours. The mortality and toxicological effects were observed for 30 h, at 6 h intervals. Important toxicological effects were observed on the larvae, such as lack of co-ordination, knock-down and paralysis followed by death, although the lethal capacity of the product after 30 h was low, with mean mortality of only 34%, 40.2%, 46% and 57.6% respectively, for the 0.5 ml/l; 1 ml/l; 2 ml/l/; and 3 ml/l dosages. There was no mortality in the control group. The results obtained did not indicate the use of this formulation, in the tested dosages, as a method of choice for R. sanguineus control. PMID- 11391438 TI - [Isolation of pathogenic bacteria in pasteurized type C milk sold in Recife City, Pernambuco, Brazil]. AB - In order to improve information about the microbiological quality of the milk commercially available in the city of Recife, 250 samples of pasteurized type-C milk and 50 samples of raw milk were analyzed for Yersinia enterocolitica and Listeria monocytogenes and verify the possible occurrence of Yersinia enterocolitica and Listeria monocytogenes. These bacteria can develop in refrigeration temperatures and are responsible for food-born diseases. Neither Y. enterocolitica nor L. monocytogenes were found in the samples analyzed. However, the presence of Y. intermedia and Y. frederiksenii was detected, these environmental species behave as opportunist pathogens. Through the methodology used for Listeria isolation, one isolate of Salmonella Montevideo was obtained from a sample of pasteurized milk and another isolated from one sample of raw milk. Besides these, several other bacteria species were found. It is likely that the large microbiota present in the samples and the procedures employed to destroy it could have hindered the isolation of Y. enterocolitica and L. monocytogenes. PMID- 11391439 TI - [Epidemiologic study of bovine and human leptospirosis in eastern Brazilian Amazon]. AB - The seroprevalence study for leptospirosis in bovines and humans was realized in family holder farms along the Transamazon Highway. The prevalence of bovine leptospirosis was 97% [90.9 - 99.5%] of farms with at least one positive animal according to microscopic agglutination test for the leptospirosis diagnostic. In 61.2% of the tested herds, the serovar hardjo was the most common, followed by the serovar bratislava (9%) and the serovar shermani (4.5%). The serologic prevalence of leptospirosis in humans was 32.8% [23.4 - 43.5%] in family groups with at least one positive individual according to microscopic agglutination test for the leptospirosis diagnostic. In 9% of family groups, the serovar bratislava was the most common, while serovar hardjo and grippotyphosa accounted for 6% and 4.5%, respectively. The impact of these results is discussed in relation to animal production and public health. Suggestions have been proposed in order to improve the situation in the region. PMID- 11391440 TI - Use of a condom in sex relations by HIV carriers. AB - The frequency with which condoms are used in sex relations by subjects with HIV was determined by interviewing 132 individuals, 82 men and 50 women, most of them from Sao Paulo state and some from other regions of the country, all of them seen at an outpatient clinic of the School of Medicine in Botucatu. The women were younger, were of lower educational level and had poorer professional qualification than men. Also, a greater proportion of women were widowed, separated or divorced. We observed that 43.9% of men and 72% of women had been contaminated by the sexual route, but only 41.2% of the men and 31.8% of the women reported the use of a condom after the diagnosis of infection, with most men and women preferring sexual abstinence. The results enable the conclusion that there is still a need to continue to provide information about the use of condoms and to guarantee their free-of-charge distribution due to the low levels of education and professional qualification of the individuals studied. The data also suggest that campaigns for the dissemination of preventive measures should consider the social and cultural differences of infected women. PMID- 11391441 TI - [Study of the vectorial competence of Lutzomyia intermedia (Lutz & Neiva, 1912) for Leishmania (Viannia) braziliensis, Vianna, 1911]. AB - This paper investigated the vectorial competence of Lutzomyia intermedia (Diptera: Psychodidae) in Vale do Ribeira (SP) to strains of Leishmania (Viannia) braziliensis (Kinetoplastida: Trypanosomatidae), by means of a search for natural infection; exposure of wild and colonized females (F1) to the lesions of experimentally infected hamsters and transmission tests by bite. The natural infection and the transmission tests were negative. In the exposures of Lu. intermedia to infected lesions we found rates of 74% (123+/166 dissected) and 70% (115+/164 dissected) for the wild and colonized females respectively. The development of the parasites was compatible with the development model of Peripilaria. The susceptibility of the tested strains associated with the epidemiological indicators contribute to the vectorial role suspicion of Lutzomyia intermedia in the studied region. PMID- 11391442 TI - [Chagasic infection prevalence in blood donors at the Regional Blood Donation Center of Iguatu]. AB - The objective of this study is to evaluate the frequency of Trypanosoma cruzi infection among blood donors at the Iguatu Regional Hemocenter, CE, 1996 - 1997, using the Enzyme-linked immunosorbent assay (ELISA) and hemagglutination passive reverse (HPR) tests. Of the 3,232 donors analyzed a total of 61 (1.9%) were seropositive for chagasic infection. The greatest number of infected donors was found in the 41-50 year age group, while the majority of donors were in the 18-30 year age group. Of the total number of donors analyzed 2,991 (92.5%) were men and 57 (1.9%) of these were seropositive. In all 1,825 (56.5 %) of the donors were from a rural area. The results showed that the ELISA test detected 49 cases of infection, but HPR only 38, thereby demonstrating that the use of two or more different tests by blood banks prevents transfusion associated Chagas' disease. PMID- 11391443 TI - [A survey of knowledge and attitudes in a population about dengue transmission- region of Campinas Sao Paulo, Brazil--1998]. AB - A survey with 502 interviews was realized in the region of Campinas (S.P.) to evaluate the community's knowledge and concepts regarding dengue: disease, vector and prevention. This information was then compared with the actual behavior towards prevention and the presence of breeding sites. Three neighborhoods of Santa Barbara D'Oeste (170,000 inhabitants) were chosen because they were located in the first municipality with autochthonous transmission, since 1995. Areas with better social and urban conditions scored higher in terms of knowledge compared with peripheral neighborhoods, even though these regions had priority in the local education program, due to case notification. However, breeding sites were found equally in both regions. A major gap between knowledge and attitude was detected in all regions. The results of the survey can help the organization of education programs to find ways to put this knowledge into practice. This instrument is useful to monitor the impact of dengue control programs. PMID- 11391444 TI - [Experimental infection in mice by Plasmodium berghei: an evidence of antiparasitic action of azithromycin]. AB - Infections of Plasmodium berghei in mice was stopped by azithromycin which was administered orally in dosages of 100mg/kg, for 28 days. This antibiotic was given since the same day that the animals were infected. The outcome suggests the necessity of more investigations on this antiparasitic activity. PMID- 11391445 TI - [AIDS and HIV infection in Brazil: a multifaceted epidemic]. AB - The HIV/AIDS epidemic is a dynamic unstable global phenomenon, constituting a veritable mosaic of regional sub-epidemics. As a consequence of the deep inequalities that exist in Brazilian society, the spread of HIV infection has revealed an epidemic of multiple dimensions undergoing extensive epidemiological transformations. Initially restricted to large urban centers and markedly masculine, the HIV/AIDS epidemic is currently characterized by heterosexualization, feminization, interiorization and pauperization. The evolution of the profile of AIDS in Brazil is above all due to the geographical diffusion of the disease from large urban centers towards medium and small municipalities in the interior, to the increase in heterosexual transmission and the persistent growth of cases among injecting drug users. The increase in transmission through heterosexual contact has resulted in substantial growth of cases among women, which has been pointed out as the most important characteristic of the epidemic's current dynamic in Brazil. PMID- 11391446 TI - Facial nerve palsy associated with leptospirosis. AB - This case report describes the findings of a 27-year-old black male from Bahia, Brazil, who developed facial palsy during the convalescence phase of leptospirosis. The patient recovered without neurological sequel. This work calls attention to a possible association between leptospirosis and facial palsy. PMID- 11391447 TI - [Benznidazol-induced jaundice]. PMID- 11391448 TI - Changes in the control program of visceral leishmaniasis in Brazil. PMID- 11391449 TI - Coffee, samba, football and. social inequalities: reflections on mortality in Sao Paulo, Brazil. PMID- 11391450 TI - The patient-physician interactions as seen by undergraduate medical students. AB - CONTEXT: The interaction between a physician and his or her patient is complex and occurs by means of technical performance and through a personal relationship. OBJECTIVE: To assess the interaction between the medical professional and his or her patient with the participation of medical students assuming a role as observers and participants in a medical appointment in an outpatient office. DESIGN: Questionnaire interview study. SETTING: General Medicine outpatient offices, Hospital das Clinicas, Faculty of Medicine, University of Sao Paulo. PARTICIPANTS: Medical students performed an ethnographical technique of observation, following 199 outpatient medical appointments with Clinical Medicine Residents. MAIN MEASUREMENTS: A questionnaire filled out by observer students measured the physician's attitudes towards patients, as well as patients' expectations regarding the appointment and his or her understanding after its completion. RESULTS: Patients showed higher enthusiasm after the appointment (4.47 +/- 0.06 versus 2.62 +/- 0.10) (mean +/- SEM), as well as some negative remarks such as in relation to the waiting time. The time spent in the consultation was 24.66 +/- 4.45 minutes (mean +/- SEM) and the waiting time was 123.09 +/- 4.91 minutes. The physician's written orientation was fairly well recalled by the patient when the doctor's letter could be previously understood. CONCLUSION: Patients benefit from physicians who keep the focus on them. In addition, this program stimulated the students for their accomplishment of the medical course. PMID- 11391451 TI - Comparison of knowledge on asthma: doctors completing internal medicine residency and doctors completing medical school. AB - CONTEXT: Asthma has been reported as a disease of increasing prevalence. OBJECTIVE: To assess the level of information and knowledge about asthma by means of a questionnaire among recent graduate physicians applying for medical residency at the Clinical Hospital of the University of Sao Paulo Medical School, Brazil. DESIGN: 14 multiple-choice questions for asthma diagnosis and management. SETTING: University of Sao Paulo Medical School (FMUSP). PARTICIPANTS: Recent graduate physicians applying for the medical residency program at FMUSP in 1999 (n = 448) and physicians that had completed 2 year of internal medicine residency (n = 92). MAIN MEASUREMENTS: We applied a questionnaire with 14 multiple-choice questions about the management of asthma based upon the Expert Panel Report 2 - Guidelines for the Diagnosis and Management of Asthma, NIH/NHLBI, 1997 (EPR-2). RESULTS: The medical residency program in Internal Medicine improved treatment skills (the ability to propose adequate therapy) when compared to medical education (a score of 57.2% versus 46.9%, P < 0.001) but not diagnosis knowledge (understanding of asthma symptoms related to medicine intake) (33.5% versus 33.3%, P = 0.94). Treatment skills were higher among physicians who received their Medical Degree (MD) from public-sponsored medical schools in comparison with those from private schools [49.7 (SE 1.17)] versus [41.8 (SE 1.63)], P < 0.001. CONCLUSION: Medical schools might consider reevaluating their programs regarding asthma in order to improve medical assistance, especially when considering the general results for residents, as they were supposed to have achieved performance after completing this in-service training. PMID- 11391452 TI - Effects of in-center daily hemodialysis upon mineral metabolism and bone disease in end-stage renal disease patients. AB - CONTEXT: Alternative hemodialysis schedules have been proposed to improve the quality of the dialysis. Nonetheless, their influence upon mineral and bone disorders is unknown. OBJECTIVE: To report the impact of a daily hemodialysis schedule upon the lesions of renal osteodystrophy. TYPE OF STUDY: Prospective non controlled study. SETTING: Public University Hospital. PARTICIPANTS: Five patients treated by daily hemodialysis for at least 24 months. INTERVENTION: Daily dialysis sessions were accomplished with non-proportional dialysis machines without an ultrafiltration control device, with blood flow of 300 ml/min, bicarbonate dialysate ([Ca]=3.5 mEq/L) at 500 ml/min, and low-flux membrane dialyzers. Sessions were started at 6:00 p.m. (except Sundays) and lasted 2 hours. MAIN MEASUREMENTS: Serum levels of Ca and P from the last 6 months on conventional hemodialysis for the same patients were used for comparison with each semester of daily hemodialysis. Bone biopsies and PTH levels were obtained at the end of the conventional hemodialysis period and then again after 2 years of daily hemodialysis. RESULTS: Mean serum calcium was significantly higher during the second and third semesters of daily dialysis [10.0 mg% (SD 0.6), and 10.0 mg% (SD 0.8), respectively] compared to standard dialysis [9.4 mg% (SD 0.8)], p < 0.05. Mean values for phosphorus were significantly lower during every semester of daily hemodialysis [6.3 mg% (SD 1.8), 5.8 mg% (SD 1.7), 6.0 mg% (SD 1.7), and 6.0 mg% (SD 1.8)] compared to standard dialysis [7.2 mg% (SD 2.7)], P < 0.05. Variations in mean Ca x P product followed the same pattern as for phosphorus [59.5 (SD 16.0), 57.1 (SD 16.3), 59.8 (SD 17.7), and 58.31 (SD 20.9) vs. 68.6 (SD 27.3), P < 0.05]. After 2 years on daily hemodialysis, 2 patients who had aplastic lesion were found to have mild bone disorder. In addition, one patient with mixed bone lesion and moderate bone aluminum accumulation had osteitis fibrosa with no aluminum. Intact PTH values at the beginning of study and after 2 years on daily hemodialysis did not differ [134 pg/ml (SD 66) vs. 109 pg/ml (SD 26), P = 0.60, respectively]. CONCLUSIONS: Patients treated using daily hemodialysis had better control of serum phosphorus and perhaps a lower risk of metastatic calcifications. Daily hemodialysis also seemed to be beneficial to low turnover bone disease and bone aluminum deposition. PMID- 11391453 TI - Value of CEA level determination in gallbladder bile in the diagnosis of liver metastases secondary to colorectal adenocarcinoma. AB - CONTEXT: The relevance of colorectal adenocarcinoma lies in its high incidence, with the liver being the organ most frequently affected by distant metastases. Liver metastases occur in 40 to 50% of patients with colorectal adenocarcinoma, accounting for approximately 80% of deaths in the first three postoperative years. Nevertheless, despite this, they are occasionally susceptible to curative treatment. OBJECTIVE: The present investigation focused on the relationship between the level of carcinoembryonic antigen (CEA) in gallbladder bile and the presence of liver metastases secondary to colorectal adenocarcinoma. DESIGN: Diagnostic test study. SETTING: Surgical Gastroenterology Discipline at the Sao Paulo Hospital, Sao Paulo, Brazil. SAMPLE: Forty-five patients with colorectal adenocarcinoma were studied, 30 without liver metastases (group I), and 15 with liver metastases (group II). Diagnosis of liver metastases was made through computed tomography, magnetic resonance imaging and computed tomography during arterial portography. Samples of peripheral blood, portal system blood, and gallbladder bile were collected from patients during the surgical procedure. A control group composed of 18 organ donors underwent the same material collection procedures. CEA level determination was made through fluoroimmunoassay. RESULTS: Mean CEA value in peripheral serum was 2.0 ng/ml (range: 0.7 to 3.8 ng/ml) in the control group, 11.4 ng/ml (range: 0.5 to 110.3 ng/ml) in group I, and 66.0 ng/ml (range: 2.1 to 670 ng/ml) in group II. In the portal system, serum mean values found were 1.9 ng/ml (range: 0.4 to 5.0 ng/ml) in the control group, 15.3 ng/ml (range: 0.8 to 133.3 ng/ml) in group I, and 70.8 ng/ml (range: 1.8 to 725 ng/ml) in group II. Mean values found in gallbladder bile were 4.1 ng/ml (range: 1.0 to 8.6 ng/ml) in the control group, 14.3 ng/ml (range: zero to 93.0 ng/ml) in group I, and 154.8 ng/ml (range: 14.0 to 534.7 ng/ml) in group II. CONCLUSIONS: The CEA level in gallbladder bile is elevated in patients with liver metastases. Determination of CEA both in peripheral serum and in gallbladder bile enabled patients with liver metastases to be distinguished from those without such lesions. The level of CEA in gallbladder bile, however, seems to lead to a more accurate diagnosis of liver metastases secondary to colorectal adenocarcinoma. PMID- 11391454 TI - Paragangliomas of the head and neck: clinical, morphological and immunohistochemical aspects. AB - CONTEXT: Protein marker positivity can assist in the definition of the therapeutic approach towards head and neck paragangliomas. The establishment of the therapeutic approach should incorporate the results of such an investigation. OBJECTIVE: To establish criteria for benignity and malignancy of vagal and jugular-tympanic paragangliomas, via the study of the relationships of sex, age, tumor size, duration of complaints, site, family history, presence of metastases, treatment, histological architecture and cell type with the immunohistochemical reactions to S100 protein, chromogranin and AgKi67. DESIGN: A retrospective study of histological and clinical records. SETTING: The Heliopolis and Oswaldo Cruz tertiary general hospitals, Sao Paulo. SAMPLE: 8 cases of head and neck paragangliomas. MAIN MEASUREMENTS: Determination of degree of positivity to paragangliomas via immunohistochemical reactions. RESULTS: 1). The protein markers for the principal cells (AgKi67 and chromogranin) were sensitive in 100% of the tumors when used together. 2). S100 protein was well identified in the cytoplasm and nucleus of sustentacular cells and underwent reduction in the neoplasias. CONCLUSIONS: Chromogranin was proven to be a generic marker for neuroendocrine tumors; S100 protein was positive in all 8 cases and the AgKi67 had low positivity in all cases. PMID- 11391455 TI - Ki-67 expression in anal intraepithelial neoplasia in AIDS. AB - CONTEXT: AIDS is one of the most important risk factors for progression and recurrence of anogenital condyloma. In a previous work, we observed that patients with warts and high-grade AIN (HAIN) had recurrences more frequently than did patients with warts without AIN. The mechanisms of this increased incidence of high-grade lesions in AIDS are not known. OBJECTIVE: We studied the expression of the proliferative marker Ki-67 by immunohistochemical methods, in specimens of anal condyloma from HIV+ patients to clarify whether its expression can be associated to the grade of AIN. DESIGN: A retrospective study of histological specimens. SETTING: University referral unit. SAMPLE: 34 patients were divided into two groups: (1) condylomas with low grade AIN (LAIN), with 25 patients; and (2) condylomas with HAIN, with 9 patients. In this latter group we examined two areas: 2A (HAIN area) and 2B (LAIN area). MAIN MEASUREMENTS: The immunohistochemical reaction for Ki-67 was done on histological sections. Slices were lightly stained with hematoxylin, to help us in Ki-67 positive cell counting. The percentage of Ki-67 marked nuclei was calculated. We applied one way variance analysis for statistics. RESULTS: The mean number of Ki-67 positive cells in group 1 was 19.68 +/- 10.99; in group 2 (area A) it was 46.73 +/- 10.409; and in area B it was 36.43 +/- 14.731. There were statistical differences between groups 1 and 2A and between groups 1 and 2B. Ki-67 positive cells predominated in the lower layer in LAIN. Positive Ki-67 cells were found in all layers in group 2A, and in group 2B they predominated in the two lower or in all layers of the epithelium. CONCLUSIONS: Our results suggest that LAIN areas (using routine staining techniques) in HAIN can have a biological behavior more similar to HAIN. PMID- 11391456 TI - Chronic activation of the innate immune system may underlie the metabolic syndrome. AB - CONTEXT: The metabolic syndrome is characterized by a clustering, in free-living populations, of cardiovascular and diabetes risk factors generally linked to insulin resistance, obesity and central obesity. Consonant with the well established inflammatory pathogenesis of atherosclerotic disease, the metabolic syndrome is now being investigated in relation to its inflammatory nature. OBJECTIVE: We present cross-sectional findings demonstrating that markers of inflammation correlate with components of the metabolic syndrome, and prospective findings of the ARIC Study indicating that markers of inflammation and endothelial dysfunction predict the development of diabetes mellitus and weight gain in adults. We present biological evidence to suggest that chronic activation of the innate immune system may underlie the metabolic syndrome, characterizing the common soil for the causality of type 2 diabetes mellitus and cardiovascular disease. CONCLUSIONS: Better understanding of the role of the innate immune system in these diseases may lead to important advances in the prediction and management of diabetes and cardiovascular disease. PMID- 11391457 TI - CMEIAS: A Computer-Aided System for the Image Analysis of Bacterial Morphotypes in Microbial Communities. AB - A major challenge in microbial ecology is to develop reliable and facile methods of computer-assisted microscopy that can analyze digital images of complex microbial communities at single cell resolution, and compute useful quantitative characteristics of their organization and structure without cultivation. Here we describe a computer-aided interactive system to analyze the high degree of morphological diversity in growing microbial communities revealed by phase contrast microscopy. The system, called "CMEIAS" (Center for Microbial Ecology Image Analysis System) consists of several custom plug-ins for UTHSCSA ImageTool, a free downloadable image analysis program operating on a personal computer in a Windows NT environment. CMEIAS uses various measurement features and two object classifiers to extract size and shape measurements of segmented, digital images of microorganisms and classify them into their appropriate morphotype. The first object classifier uses a single measurement feature to analyze relatively simple communities containing only a few morphotypes (e.g., regular rods, cocci, filaments). A second new hierarchical tree classifier uses an optimized subset of multiple measurement features to analyze significantly more complex communities containing greater morphological diversity than ever before possible. This CMEIAS shape classifier automatically categorizes each cell into one of 11 predominant bacterial morphotypes, including cocci, spirals, curved rods, U-shaped rods, regular straight rods, unbranched filaments, ellipsoids, clubs, rods with extended prostheca, rudimentary branched rods, and branched filaments. The training and testing images for development and evaluation of the CMEIAS classifier were obtained from 1,937 phase-contrast grayscale digital images of various diverse communities. The CMEIAS shape classifier had an accuracy of 96.0% on a training set of 1,471 cells and 97.0% on a test set of 4,270 cells representing all 11 bacterial morphotype classes, indicating that accurate classification of rich morphological diversity in microbial communities is now possible. An interactive edit feature was added to address the main sources of error in automatic shape classification, enabling the operator to inspect the assigned morphotype of each bacterium based on visual recognition of its distinctive pseudocolor, reassign it to another morphotype class if necessary, and add up to five other morphotypes to the classification scheme. The shape classifier reports on the number and types of different morphotypes present and the abundance among each of them, thus providing the data needed to compute the morphological diversity within the microbial community. An example of how CMEIAS can augment the analysis of microbial community structure is illustrated by studies of morphological diversity as an indicator of dynamic ecological succession following a nutrient shift-up perturbation in two continuously fed, anaerobic bioreactors with morphologically distinct start communities. Various steps to minimize the limitations of computer-assisted microscopy to classify bacterial morphotypes using CMEIAS are described. In summary, CMEIAS is an accurate, robust, flexible semiautomatic computing tool that can significantly enhance the ability to quantitate bacterial morphotype diversity and should serve as a useful adjunct to the analysis of microbial community structure. This first version of CMEIAS will be released as free, downloadable plug-ins so it can provide wide application in studies of microbial ecology. PMID- 11391458 TI - Quantifying Population Dynamics Based on Community Structure Fingerprints Extracted from Biosolids Samples. AB - This paper presents a standardized metric for quantifying the rate change in community structure of complex mixed microbial cultures such as those maintained in biological wastewater treatment systems. Quantifying the stability of microbial community structures is a first step toward more aggressive monitoring and control of biological systems for greater reliability in contaminant removal. Statistical analysis of compositions that uniquely specify the balance of populations of species in a mixed culture sample can be used to specify a biosolids community state as a unique position in an orthogonal coordinate system. Changes in biosolids state are observable as a trajectory within this coordinate space, and the rate of passage along this trajectory relates to the population dynamics. The geometric interpretation and the statistical analysis methods necessary for the proposed calculation methodology are introduced by way of simulated case studies with a simple model system. With the example of this model system, concepts of changing microbial community shape and size are contrasted. The rate change in community structure is defined geometrically in terms of a rate change in relative proportions of the characteristic community shape at constant biomass. A change in biomass is defined as the rate change in the quantity of biosolids at constant shape. The method robustness with respect to random measurement error was also demonstrated using the model system. The potential applications of the approach are presented with experimental data of microbial fatty acid compositions extracted from samples during the operation of bench-scale bioreactors degrading contaminants found in pulp mill wastewater. Scaling the level of population dynamics with a metric that is independent of chemotypic content presents a standard for direct comparisons of community responses between distinct cultures and experiments. PMID- 11391459 TI - Microbial Competition in Reactors with Wall Attachment. AB - Competition for nutrient and the ability of bacteria to colonize the gut wall are factors believed to play a role in the observed stability of the indigenous microbiota of the mammalian large intestine. These factors were incorporated into the two-strain continuous-stirred tank reactor (CSTR) model formulated and numerically investigated by Freter et al. In their model simulations, the reactor is parameterized using data for the mouse intestine. An invading bacterial strain is introduced into a CSTR that has already been colonized by a resident strain. The two strains compete for a single growth-limiting nutrient and for limited adhesion sites on the wall of the reactor. The mathematical model described in this paper is motivated in part by the CSTR model, but is based on the plug flow reactor (PFR). Parameter values and initial conditions are chosen so that the numerical performance of the PFR can be compared to that of the CSTR. In simulations bearing a remarkable qualitative and quantitative resemblance to those of the CSTR it is found that the invader is virtually eliminated, despite the fact that it has uptake rate and affinity for the wall identical to those of the resident. The PFR model is then parametrized using data for the human large intestine, and the two-strain simulations are repeated. Though obvious quantitative differences are noted, the more important qualitative outcome is preserved. It is also found that when three strains compete for a single nutrient and for adhesion sites there exists a steady-state solution characterized by the segregation of the bacterial strains into separate nonoverlapping segments along the wall of the reactor. PMID- 11391460 TI - Persistence and Distribution of Wild-Type and Recombinant Nucleopolyhedroviruses in Soil. AB - Persistence of recombinant and wild-type nucleopolyhedroviruses (NPV) was compared in field and laboratory microcosm experiments. Horizontal and vertical distribution of the viruses also was monitored in the field agricultural soil. Mixed populations of the bollworm, Helicoverpa zea, and tobacco budworm, Heliothis virescens, in cotton were sprayed five times during a growing season with wild-type H. zea NPV (HzSNPV.WT) or with a genetically modified H. zea NPV expressing an insect-specific neurotoxin (HzSNPV.LqhIT2). HzNPV.WT accumulated 2.3 times as many occlusion bodies (OB) as HzSNPV.LqhIT2 in soil by the end of the growing season in October 1997. Both NPVs were detected at all soil depths down to 26-35 cm. Both NPVs were randomly distributed among 0-2 cm soil samples throughout the plots according to analysis with Taylor's power law. By 4 August 1998, soil concentration of HzSNPV.WT was only 11-13 OB/g at depths from 0 to 14 cm, and the wild-type virus was not detected below 14 cm. HzSNPV.LqhIT2 was detected only in trace amounts at 0-2 cm at this time. Neither NPV was detected in bioassays of cotton leaves nor in insects sampled from the plots in 1998. Viral persistence also was monitored in laboratory soil microcosms. Three viruses wild-type Autographa californica NPV (AcNPV.WT), A. californica NPV expressing a scorpion toxin (AcNPV.AaIT), and A. californica NPV expressing juvenile hormone esterase (AcNPV.JHE-S201G)-were introduced into soil microcosms by each of two methods, in water suspension or in host cadavers, for a total of six treatments plus controls. After 17 months, the number of viable OB remaining did not differ among the treatments. The results indicate that the only differences in soil populations of wild-type versus recombinant NPVs are due to the greater amounts of the wild-type viruses that accumulate, probably because they have a greater capacity to replicate in the host insect population. PMID- 11391461 TI - Motility Responses and Desiccation Survival of Zoospores from the Actinomycete Kineosporia sp. Strain SR11. AB - During investigations into the effects of water availability on the ecology of leaf litter actinomycetes, a motile-spored strain (SR11) was isolated and identified as a Kineosporia sp. Observations of spore motility revealed they could achieve speeds up to 160 mm s?1. Chemotaxis was not observed toward several organic compounds, although K, Mg, and Ca salts of phosphate, sulfate, and halides elicited a positive response. The inability of all the corresponding Na salts, and KNO3, to act as attractants suggested that chemoattraction of SR11 spores toward inorganic compounds may be dependent on specific cations and anions. An unusual motility response to CO2 was also observed and a laminar-flow microchamber was designed and constructed to investigate this phenomenon. Increasing the CO2 concentration by as little as 0.3% (v/v) caused a rapid, synchronous arrest of spore motility. A 1% (v/v) decrease in CO2 concentration caused a similar response, although the duration of the pause was much longer. The survival responses of strain SR11 spores were observed following desiccation at various water potentials. At the highest water potential tested (-9 MPa), spore viability declined minimally during the first 24 hr but continued to decrease over 10 days. In contrast, desiccation at -87 MPa and below resulted in a 50% loss in viability within 2 hr and subsequent survival at this level of viability. Continued desiccation at -302 MPa indicated that spores were capable of long-term survival at low water potentials. Many of these responses are likely to be significant in determining the ecology of these motile-spored actinomycetes in leaf litter. PMID- 11391462 TI - The Influence of Competition between Tropical Fungi on wood colonization in Streams. AB - Wood blocks inoculated with fungi were placed in a stream in order to investigate their interactions with the native mycota. Wood blocks were inoculated with single or paired isolates of tropical fungi, whereas no fungi were inoculated onto controls. After 3 months, the wood blocks were collected and the number of newly colonizing fungi were identified. More fungi were found on uninoculated than on wood blocks pre-inoculated with Chaetosphaeria sp. Neither Nais aquatica or Annulatascus velatisporus inhibited colonization when inoculated singly. However, in combination, they inhibited colonization by other fungi, indicating an interaction between the two species. Inoculated wood blocks were generally first colonized by hyphomycetes, with a low occurrence of ascomycetes. The study also showed that autoclaving wood for use in experiments affects the outcome of the experiments, i.e., affects the structure of the fungal community. PMID- 11391463 TI - The Diversity of Archaea and Bacteria in Association with the Roots of Zea mays L. AB - The diversity of bacteria and archaea associating on the surface and interior of maize roots (Zea mays L.) was investigated. A bacterial 16S rDNA primer was designed to amplify bacterial sequences directly from maize roots by PCR to the exclusion of eukaryotic and chloroplast DNA. The mitochondrial sequence from maize was easily separated from the PCR-amplified bacterial sequences by size fractionation. The culturable component of the bacterial community was also assessed, reflecting a community composition different from that of the clone library. The phylogenetic overlap between organisms obtained by cultivation and those identified by direct PCR amplification of 16S rDNA was 48%. Only 4 bacterial divisions were found in the culture collection, which represented 27 phylotypes, whereas 6 divisions were identified in the clonal analysis, comprising 74 phylotypes, including a member of the OP10 candidate division originally described as a novel division level lineage in a Yellowstone hot spring. The predominant group in the culture collection was the actinobacteria and within the clone library, the a-proteobacteria predominated. The population of maize-associated proteobacteria resembled the proteobacterial population of a typical soil community within which resided a subset of specific plant-associated bacteria, such as Rhizobium- and Herbaspirillum-related phylotypes. The representation of phylotypes within other divisions (OP10 and Acidobacterium) suggests that maize roots support a distinct bacterial community. The diversity within the archaeal domain was low. Of the 50 clones screened, 6 unique sequence types were identified, and 5 of these were highly related to each other (sharing 98% sequence identity). The archaeal sequences clustered with good bootstrap support near Marine group I (crenarchaea) and with Marine group II (euryarchaea) uncultured archaea. The results suggest that maize supports a diverse root associated microbial community composed of species that for the first time have been described as inhabitants of a plant-root environment. PMID- 11391464 TI - Grazing Characteristics and Growth Efficiencies at Two Different Temperatures for Three Nanoflagellates Fed with Vibrio Bacteria at Three Different Concentrations. AB - Small inocula of one of the flagellates Paraphysomonas imperforata, Pteridomonas danica, and Cafeteria roenbergensis were added to suspensions of the bacterium Vibrio natriegens at each of three concentrations between 107 and 108 cells ml-1 and incubated at each of the temperatures 10 degrees C and 25 degrees C. Samples were taken at intervals for counting the flagellates and bacteria to determine the timing of the maximum of flagellate numbers and the concentrations at that time. Measurements of the protein concentration of the suspensions during incubation were used to determine the gross growth efficiency (GGE) or yield of flagellate grazing in each experiment. The most effective grazer was Pteridomonas, followed by Paraphysomonas, with Cafeteria being least effective, as judged by the threshold bacterial concentrations at which flagellate multiplication ceased, which were about 2 x 105, 2 x 106, and 2 x 107, respectively, and by the finding that Pteridomonas consumed 99%, Paraphysomonas about 95%, and Cafeteria only 60-70% of the available bacteria in the experiments. Peak concentrations of flagellates were reached later at the lower temperature, but the numbers of flagellates produced and of bacteria eaten were of a similar order at the two temperatures and the GGE was only slightly higher at the lower temperature. The time taken to reach peak flagellate numbers changed little with a threefold increase in bacterial concentrations, but the GGE increased and the numbers of bacteria eaten to produce one flagellate decreased when the bacterial concentration was increased. The three flagellates show clear evidence of niche specialization in differences in thresholds of bacterial prey concentration. PMID- 11391465 TI - Influence of Initial C/N Ratio on Chemical and Microbial Composition during Long Term Composting of Straw. AB - Shredded straw of Miscanthus was composted in 800-L boxes with different amounts of pig slurry added as nitrogen source. The impact of the different initial C/N ratios (11, 35, 47, 50, and 54) on the composting process and the end product was evaluated by examining chemical and microbiological parameters during 12 months of composting. Low initial C/N ratios caused a fast degradation of fibers during the first three months of composting (hemicellulose: 50-80%, cellulose: 40-60%), while high initial C/N ratios resulted in 10-20% degradation of both hemicellulose and cellulose. These differences were reflected in the microbial biomass and respiration, which initially were higher in low C/N treatments than in high C/N treatments. After 12 months of composting, this situation was reversed. Composts with high initial C/N ratios had high microbial biomass (15-20 mg ATP g-1 OM) and respiration rates (200 mg CO2 h-1 g-1 OM) compared to treatments with low initial C/N ratios (less than 10 mg ATP g-1 OM and 25 mg CO2 h-1 g-1 OM). This could be explained by the microorganisms being nitrogen limited in the high C/N ratio treatments. In the low C/N ratio treatments, without nitrogen limitation, the high activity in the beginning decreased with time because of exhaustion of easily available carbon. Different nitrogen availability was also seen in the nitrification patterns, since nitrate was only measured in significant amounts in the treatments with initial C/N ratios of 11 and 35. The microbial community structure (measured as phospholipid fatty acid, PLFA, profile) was also affected by the initial C/N ratios, with lower fungal/bacterial ratios in the low compared to the high C/N treatments after 12 months of composting. However, in the low C/N treatments higher levels of PLFAs indicative of thermophilic gram-positive bacteria were found compared to the high C/N treatments. This was caused by the initial heating phase being longer in the low than in the high C/N treatments. The different fungal/bacterial ratios could also be explained by the initial heating phase, since a significant correlation between this ratio and heat generated during the initial composting phase was found. PMID- 11391466 TI - Expression of the ACC Deaminase Gene fromEnterobacter cloacae UW4 in Azospirillum brasilense. AB - The ACC deaminase structural gene (acdS) from Enterobacter cloacae UW4 was cloned in the broad host range plasmid pRK415 under the control of the lac promoter and transferred into Azospirillum brasilense Cd and Sp245. A. brasilenseCd and Sp245 transformants showed high ACC deaminase activity, similar to that observed in Enterobacter cloacae UW4. The expression of ACC deaminase improved the existing growth promoting activity of Azospirillum. The roots of tomato and canola seedlings were significantly longer in plants inoculated with A. brasilense Cd transformants than those in plants inoculated with the nontransformed strains of the same bacterium. In the case of wheat seedlings, inoculation with A. brasilense Cd transformants did not promote root growth. The difference in plant response (canola and tomato versus wheat) is attributed to the greater sensitivity of canola and tomato plants to ethylene as compared to wheat plants. PMID- 11391467 TI - A rapid method for determining the tuberculocidal activity of liquid chemical germicides. AB - A rapid, quantitative method has been developed for determining the tuberculocidal activity of liquid chemical germicides. In this method, a test strain of Mycobacterium bovis that carries the firefly luciferase gene is exposed to a germicide, and the surviving bacteria are detected by bioluminescence. The tuberculocidal activities of five commercially available glutaraldehyde-based disinfectants were tested, and all reduced the number of surviving mycobacteria by greater than five orders of magnitude. In contrast, a phenol-based disinfectant with tuberculocidal claims gave less than one order of magnitude reduction of the test organism. With this method for determining tuberculocidal activity, results can be obtained in less than one day, compared with weeks or months for the standard tuberculocidal assays. PMID- 11391468 TI - Monitoring of bio-oxidation process of ferrous ion by using piezoelectric impedance analysis. AB - A new method of monitoring the bio-oxidation process of ferrous ion in the presence of Thiobacillus ferroxidans was proposed by piezoelectric impedance analysis. The time courses of the responses of impedance parameters for a quartz crystal in a culture system were simultaneously obtained and discussed. It was found that the frequency shift response originates mainly from the adsorption of bacterial metabolites on the surface of gold electrode. Experiments also examined the effect of culture temperature on the bio-oxidation process. Combined with the growth situation of the bacterium, an impedance response model reflecting the process was established. By fitting Delta f vs. time curves toward the proposed model, we obtained and discussed the bacterial growth parameters. The results showed that the proposed method could provide real time and multidimensional information to monitoring of the bio-oxidation process. PMID- 11391469 TI - An E. coli 5S rRNA deletion mutant useful for the study of 5S rRNA structure/function relationships. AB - We report on the construction of a novel strain of E. coli that can be useful for studies on the structure/function relationship of 5S rRNAs. The bacterial strain is deficient in six of the eight naturally occurring 5S rRNA genes (operons B, D, H, G, E) and demonstrates a greatly reduced growth rate that can be compensated by the plasmid-encoded expression of 5S rRNA. The relatively large difference in growth rate between compensated and non-compensated mutants provides the basis for a quick and simple assaying system for both the evaluation and mass screening of divergent 5S rRNA sequences for function. We describe the construction of the 5S rRNA deletion mutant BDHGE and characterize the usefulness and limitations of the system for evaluating structure/function relationships of 5S rRNA sequence. PMID- 11391470 TI - The Saccharomyces cerevisiae MLF6/YPL244C gene, which encodes a possible yeast homologue of mammalian UDP-galactose transporter, confers resistance to the immunosuppressive drug, leflunomide. AB - The immunosuppressant leflunomide (LFM) inhibits the growth of cytokine stimulated proliferation of lymphoid cells in vitro and also inhibits the growth of eukaryotic microorganism, Saccharomyces cerevisiae. As a first step to elucidate the molecular mechanism of LFM action in human, yeast gene which suppresses the anti-proliferative effect when in increased copy number was cloned and designated MLF6 for multicopy suppressor of LFM sensitivity. DNA sequencing analysis revealed that the MLF6 gene is identical to the YPL244C gene which encodes a possible yeast homologue of human UDP-galactose transporter. The disruption of the MLF6 gene increased the sensitivity of yeast cells to the drug. PMID- 11391471 TI - Identification of photolabile outer membrane proteins of Porphyromonas gingivalis. AB - As the prevalence of antibiotic-resistant strains of bacteria increases, novel ways of treating infections need to be developed. This is particularly pertinent with respect to the periodontal diseases--the most common chronic bacterial infections of man. The use of a photosensitizer in combination with red light has been demonstrated to be effective in killing several human pathogens, including the oral bacterium, Porphyromonas gingivalis, a major pathogen in periodontitis. Killing was associated with alterations in the molecular masses of several outer membrane and plasma membrane proteins and these may be therapeutic targets for photodynamic therapy and other antimicrobial approaches. To identify these photolabile proteins, we have used a panel of monoclonal antibodies raised to whole P. gingivalis. A number of the antibodies recognized various photolabile proteins. Using a combination of Western blotting and protein sequencing the predominant photolabile proteins in P. gingivalis have been identified as the major secreted/cell surface proteases--Lys and Arg gingipain. PMID- 11391472 TI - Cell-associated and extracellular proteinases in Blastocrithidia culicis: influence of growth conditions. AB - The proteinase profile of Blastocrithidia culicis was analyzed, as well as how different growth conditions influenced its expression by gelatin-SDS-PAGE and the use of specific proteinase inhibitors. Multiple cell-associated proteinases with molecular masses corresponding to 33, 55, 60 kDa (cysteine proteinases) and 77, 80, 90, and 108 kDa (metalloproteinases) were detected using these methods. All the metalloproteinases identified were partitioned into the detergent phase after Triton X-114 extract, suggesting that they are membrane-bound, while all cysteine proteinases were partitioned into the aqueous phase. The proteolytic zymograms were similar when three different media were used for variable times of incubation. However, few quantitative and qualitative changes were observed. The secreted proteinase profile showed an heterogeneous pattern of enzymatic activities whose expression was dependent on time of growth and medium composition. However, the extracellular proteinase activities were abolished by 1,10-phenanthroline, suggesting that all of them are zinc-metalloproteinases. PMID- 11391473 TI - Purification and kinetic properties of betaine-homocysteine methyltransferase from Aphanothece halophytica. AB - Betaine-homocysteine methyl transferase (BHMT) from Aphanothece halophytica was purified to homogeneity by hydroxyapatite, DEAE-Sepharose CL-6B and Sephadex G 200 column chromatography. A 24-fold purification and 11% overall yield were achieved with a specific activity of 595 nmol h(-1) mg(-1). The subunit molecular weight was determined to be 45 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and the native enzyme was found to have a molecular weight of 350 kDa, suggesting an octameric structure of the enzyme. The enzyme shows optimum activity at 37 degrees C, pH 7.5. The apparent Km values for glycinebetaine and L-homocysteine were 4.3 mM and 1.3 mM, respectively. The enzyme was 70% inactivated by 5 mM dimethylglycine whereas the same concentration of sarcosine slightly inactivated the enzyme. Two analogs of glycinebetaine were also tested for enzyme inactivation and it was found that 5 mM choline inactivated 60% of the enzyme activity and 2.5 mM betaine aldehyde completely abolished the enzyme activity. NaCl at 200 mM or higher also completely inactivated the enzyme. PMID- 11391474 TI - Highly toxic and broad-spectrum insecticidal Bacillus thuringiensis engineered by using the transposon Tn917 and protoplast fusion. AB - The chromosome of the Bacillus thuringiensis strain S184 that was toxic against the third instar larvae of Spodoptera litura with the LC(50) of 9.74 microg/ml was successfully integrated into two genes of cyt1Aa and cry11Aa using the transposon Tn917, yielding the primary engineered strain TnX. The strain TnX was highly toxic to the third instar larvae of Culex pipiens fatigans with the LC(50) of 5.12 ng/ml which was 1.82-fold higher than that of B. thuringiensis subsp. israelensis, but lowly toxic to lepidopterous larvae. By the protoplast fusion of the strain TnX and the strain S184-Tet(r) (resistance to tetracycline), the target engineered strain TnY was obtained. Against the third instar larvae of S. litura, the strain TnY LC(50) was of 4.68 micro g/ml and increased by 2.08-fold in comparison with the parent strain S184. Against the third instar larvae of C. pipiens fatigans, the strain TnY LC(50) was of 103.20 ng/ml. The two target genes of cyt1Aa and cry11Aa integrated into the chromosome were extremely stable and had little possibility of a second transposition. It was unclear whether some factors existing in the parent strain, S184, contributed to the high toxicity of the strains TnX and TnY. PMID- 11391475 TI - UV-induced increase in RNA polymerase activity in Xanthomonas oryzae pathovar oryzae. AB - UV radiation is thought to inhibit transcriptional elongation, as a result of the formation of pyrimidine dimers in the DNA template, as well as to activate specific transcription factors. However, the effect of UV radiation on the enzymatic activity of RNA polymerase has remained unknown. With the use of an in vitro assay, UV irradiation of Xanthomonas oryzae pathovar oryzae has now been shown to increase RNA polymerase activity. This effect was maximal at a UV dose of approximately 12 J m(-2) and at approximately 60 min after irradiation. It was also not inhibited by pretreatment of cells with chloramphenicol, an inhibitor of protein synthesis. Immunoprecipitation with antibodies to the RNA polymerase core enzyme revealed that exposure of the bacterial cells to UV radiation induced the association of the core enzyme with a protein of approximately 29 kDa. These results demonstrate that UV radiation increases the activity of RNA polymerase, and they suggest that this effect may be related to the repair of DNA damage. PMID- 11391476 TI - Vancomycin resistance among strains of Staphylococcus epidermidis: effects on adherence to silicone. AB - Nosocomial device-related infections with Gram-positive cocci and their resistance to vancomycin are of increasing occurrence. We examined clinical isolates of relatively avirulent coagulase-negative staphylococci for their resistance to vancomycin and for their capabilities to adhere in vitro to medical grade silicone. Vancomycin resistance was found in 9 of 20 isolates, but there was no correlation between adherence capacity to silicone in the absence of vancomycin and vancomycin resistance for a given strain. Vancomycin in the medium, adsorbed to the surface of medical grade silicone or adsorbed on nongrowing cells, reduced adherence of representative Staphylococcus epidermidis to medical grade silicone. PMID- 11391477 TI - Listeria monocytogenes lineage group classification by MAMA-PCR of the listeriolysin gene. AB - Nucleotide sequence differences within several virulence genes, including the listeriolysin O (hly) gene, are associated with three evolutionary lineage groups of Listeria monocytogenes. Because the ability of L. monocytogenes to cause disease may vary by evolutionary lineage group, rapid discrimination among the three lineage types may be important for estimating pathogenic potential. A Mismatch Amplification Mutation Assay (MAMA) was developed and used to rapidly screen and characterize L. monocytogenes isolates with regard to lineage type. A standard PCR amplified a 446-bp region within the hly gene with all three L. monocytogenes lineage genotypes. MAMA primers to four different sites within this region of the hly gene were designed to amplify under the same PCR conditions and generated amplicons, the size of which depended on the isolate genotype. Ninety seven L. monocytogenes isolates were screened. All isolates, except ATCC 19116, could be classified by MAMA PCR as one of the three hly genotypes. Overall, 56, 36, and 4 of the 97 isolates tested were type 1, 2, or 3 respectively. Among the 26 patient isolates, 85%, 15%, and 0% were type 1, 2, or 3 respectively; for the 60 food isolates, 54% were type 1, 43% were type 2, and 3% were type 3. The combination of these MAMA PCR analyses provides a rapid method to screen and categorize L. monocytogenes isolates because of conserved nucleotide differences within the hly gene. PMID- 11391478 TI - Isolation of stable hemolysin and catalase variants of Staphylococcus aureus S6C includes one with an exoprotein-deficient phenotype. AB - Previous studies suggested that the exoprotein-deficient phenotype of a Delta 1058::Tn551 insertion/deletion mutant of Staphylococcus aureus S6C was not owing to the insertion/deletion event, but instead was owing to the inherent instability of the agrC gene during transduction of the Delta 1058::Tn551 region into S6C. The purpose of the following study was to examine S6C as a potential source of exoprotein-deficient mutants that would account for their appearance after transposition and transduction. Four stable variants of S6C were isolated that differed in their hemolysin and catalase activities. Surprisingly, the agr regulatory molecule, RNAIII, was undetectable in one of these variants, which most likely accounted for the exoprotein-deficient phenotype of this variant. When the original Delta 1058::Tn551 mutation was transduced into the hemolytic, catalase-positive variant of S6C, none of the transductants exhibited an exoprotein-deficient phenotype. These data suggest that, while the exoprotein deficient phenotype of the S6C variant is most likely due to mutations in the agr regulatory system, these mutations are not caused by the transduction of the Delta 1058::Tn551 region into S6C, but instead already exist in an exoprotein deficient variant of S6C. PMID- 11391479 TI - Comparative sensitivity to UV-B radiation of two Bacillus thuringiensis subspecies and other Bacillus sp. AB - Susceptibility of Bacillus thuringiensis spores and toxins to the UV-B range (280 -330 nm) of the solar spectrum reaching Earth's surface may be responsible for its inactivation and low persistence in nature. Spores of the mosquito larvicidal B. thuringiensis subsp. israelensis were significantly more resistant to UV-B than spores of the lepidopteran-active subsp. kurstaki. Spores of subsp. israelensis were as resistant to UV-B as spores of B. subtilis and more resistant than spores of the closely related B. cereus and another mosquito larvicidal species B. sphaericus. Sensitivity of B. thuringiensis subsp. israelensis spores to UV-B radiation depended upon their culture age; 24-h cultures, approaching maximal larvicidal activity, were still sensitive. Maximal resistance to UV-B was achieved only at 48 h. PMID- 11391480 TI - Arsenite-induced multiple antibiotic resistance phenotype in environmental isolates of Yersinia enterocolitica. AB - Minimal inhibitory concentrations (MICs) of five antibiotics namely amikacin, gentamicin, tetracycline, ciprofloxacin, and nitrofurantoin for pork isolates of Yersinia enterocolitica increased two- to eightfold after bacteria were grown in the presence of 5 mm arsenite. For Y. enterocolitica isolates obtained from wastewater (sewage effluents), an unequivocal increase in MICs was seen with amikacin and gentamicin. No change was discernible in the outer-membrane proteins after isolates were grown in the presence of arsenite. PMID- 11391481 TI - Homocysteine metabolism in children with Down syndrome: in vitro modulation. AB - The gene for cystathionine beta-synthase (CBS) is located on chromosome 21 and is overexpressed in children with Down syndrome (DS), or trisomy 21. The dual purpose of the present study was to evaluate the impact of overexpression of the CBS gene on homocysteine metabolism in children with DS and to determine whether the supplementation of trisomy 21 lymphoblasts in vitro with selected nutrients would shift the genetically induced metabolic imbalance. Plasma samples were obtained from 42 children with karyotypically confirmed full trisomy 21 and from 36 normal siblings (mean age 7.4 years). Metabolites involved in homocysteine metabolism were measured and compared to those of normal siblings used as controls. Lymphocyte DNA methylation status was determined as a functional endpoint. The results indicated that plasma levels of homocysteine, methionine, S adenosylhomocysteine, and S-adenosylmethionine were all significantly decreased in children with DS and that their lymphocyte DNA was hypermethylated relative to that in normal siblings. Plasma levels of cystathionine and cysteine were significantly increased, consistent with an increase in CBS activity. Plasma glutathione levels were significantly reduced in the children with DS and may reflect an increase in oxidative stress due to the overexpression of the superoxide dismutase gene, also located on chromosome 21. The addition of methionine, folinic acid, methyl-B(12), thymidine, or dimethylglycine to the cultured trisomy 21 lymphoblastoid cells improved the metabolic profile in vitro. The increased activity of CBS in children with DS significantly alters homocysteine metabolism such that the folate-dependent resynthesis of methionine is compromised. The decreased availability of homocysteine promotes the well established "folate trap," creating a functional folate deficiency that may contribute to the metabolic pathology of this complex genetic disorder. PMID- 11391484 TI - [Quantitative evaluation of disorders of coordination in patients with Cuban type 2 spinocerebellar ataxia]. AB - INTRODUCTION: Implementation of quantitative techniques to study disorders of coordination of movements in hereditary ataxias permits an objective description of the disease. OBJECTIVES: To make an objective evaluation of the main disorders of coordination in patients with ataxia and compare healthy persons with those affected. PATIENTS AND METHODS: We made a prospective transverse study of 43 normal persons and 50 patients with type 2 spinocerebellar ataxia (SCA2). In all cases we made a qualitative examination of Romberg's sign in two stages to increase the sensitivity. We observed how long the person could keep his balance or whether he swayed and fell. The finger-nose test and diadochokinesia were also quantitatively analyzed using a specially designed system connected to a computer which permitted quantitative analysis. RESULTS: We showed that the patients with SCA2 swayed when standing upright. However, they swayed and fell more when the test became more sensitive. This showed loss of postural sense. The maximum variables of the period and standard deviation were increased, whilst the effectiveness was significantly reduced in the group of patients (finger-nose test). Quantitative examination of diadochokinesia showed increased values for the maximum period, standard deviation and integral. CONCLUSIONS: The techniques developed allow measurement of the main disorders of coordination in patients with SCA2, help to differentiate affected persons from healthy ones and are useful for the detection of moderate changes in severity during progression of the disorder. PMID- 11391482 TI - Etiological point mutations in the hereditary multiple exostoses gene EXT1: a functional analysis of heparan sulfate polymerase activity. AB - Hereditary multiple exostoses (HME), a dominantly inherited genetic disorder characterized by multiple cartilaginous tumors, is caused by mutations in members of the EXT gene family, EXT1 or EXT2. The corresponding gene products, exostosin 1 (EXT1) and exostosin-2 (EXT2), are type II transmembrane glycoproteins which form a Golgi-localized heterooligomeric complex that catalyzes the polymerization of heparan sulfate (HS). Although the majority of the etiological mutations in EXT are splice-site, frameshift, or nonsense mutations that result in premature termination, 12 missense mutations have also been identified. Furthermore, two of the reported etiological missense mutations (G339D and R340C) have been previously shown to abrogate HS biosynthesis (McCormick et al. 1998). Here, a functional assay that detects HS expression on the cell surface of an EXT1 deficient cell line was used to test the remaining missense mutant exostosin proteins for their ability to rescue HS biosynthesis in vivo. Our results show that EXT1 mutants bearing six of these missense mutations (D164H, R280G/S, and R340S/H/L) are also defective in HS expression, but surprisingly, four (Q27K, N316S, A486V, and P496L) are phenotypically indistinguishable from wild-type EXT1. Three of these four "active" mutations affect amino acids that are not conserved among vertebrates and invertebrates, whereas all of the HS-biosynthesis null mutations affect only conserved amino acids. Further, substitution or deletion of each of these four residues does not abrogate HS biosynthesis. Taken together, these results indicate that several of the reported etiological mutant EXT forms retain the ability to synthesize and express HS on the cell surface. The corresponding missense mutations may therefore represent rare genetic polymorphisms in the EXT1 gene or may interfere with as yet undefined functions of EXT1 that are involved in HME pathogenesis. PMID- 11391485 TI - [Epidemiology of Cuban hereditary ataxia]. AB - INTRODUCTION: The hereditary ataxias in Cuba make up the highest concentration of these patients in the world. The spinocerebellar ataxia type 2 (SCA2) molecular form is predominant. OBJECTIVES: To determine the prevalence and incidence of hereditary ataxias. PATIENTS AND METHODS: We made a descriptive study of 440 patients and 1,633 members of families at risk from this disorder in the province of Holguin. We calculated the prevalence rate and incidence. RESULTS: The rate of prevalence of patients in the province is 43 cases per 100,000 inhabitants; the highest rate was 503 cases per 100,000 inhabitants in part of the municipality of Baguanos. The age group that was most affected was that of 30-39 years, with a prevalence of 63.97 cases per 100,000 inhabitants. The rural population showed the highest incidence (62.04 cases per 100,000 inhabitants). The risk of members of affected families showing the disorder was 159.33 cases per 100,000 inhabitants in this province. The highest incidence was 18.08 cases per 100,000 inhabitants in Cacocum, where the incidence in the province was 4.39. CONCLUSIONS: On average the disorder passes from one state to the next every year, which suggests that the extent of the disorder worsens with time. The prevalence and incidence are the highest in the world. This together with the dominant pattern of inheritance, the effect of anticipation and inexorably progressive course of the disorder shows the serious health problem that affects the Eastern region of Cuba. PMID- 11391486 TI - [Prevalence of dissocial conduct disorder in adolescents using an epidemiological diagnostic questionnaire]. AB - INTRODUCTION: Prevalence of Conduct Disorder has been calculated between 4 to 10%. OBJECTIVE: To estimate the prevalence CD in Colombian adolescents, using a rating scale by self report. PATIENTS AND METHODS: 190 male adolescents from secondary school population, 12 to 16 year-old, were stratified and selected in a randomizing procedure, from Medellin, Colombia. A checklist including 14 diagnostic symptoms taken from the DSM-IV Criterion A for CD was constructed, with a scale of 0 = never to 3 = almost always (Reliability alpha coefficient: 0.86). Prevalences were calculated as follow: (1) codifying as 'Probable TDC' a T score over 60; (2) codifying as Adolescent Antisocial Behavior (AAB) having three or more answers qualified with 1 or more points, and (3) codifying as CD obtaining three or more answers qualified with 2 or 3 points. RESULTS: The most prevalent CD symptoms were 'staying out at night before 13 year-old' (10.5%), 'having been cruel to animals' (8.4%), 'having been cruel to people' (7.4%), 'having broken into someone else's house or car' (7.3%), and 'using weapon or other objects that can cause serious physical harm to others' (6.9%). The prevalence of 'Probable CD' was 13.7%; prevalence of AAB was 56.8% and prevalence of CD was 8.4%. It was not found significant differences between socioeconomic strata. CONCLUSION: The prevalence of CD in the studied sample was 8.4%, independent of the socioeconomic strata. PMID- 11391483 TI - Friedreich ataxia: from GAA triplet-repeat expansion to frataxin deficiency. PMID- 11391487 TI - [An atypical rhabdoid/teratoid tumor. A presentation of three cases and review of the literature]. AB - INTRODUCTION: The atypical rhabdoid/teratoid tumor of the central nervous system (CNS) has emerged from a variant of medulloblastoma to become a distinctive clinico-pathological condition over the past ten years. This is more than simply nosological importance, since the five year survival expected in medulloblastomas is considerably better than the 11 months (average) seen in rhabdoid/teratoid tumors. OBJECTIVE: To describe the main clinico-pathological characteristics of a recently recognized condition, emphasizing its polymorphism and differential diagnosis. PATIENTS AND METHODS: We report the clinical, radiological, cytohistopathological and immunohistopathological aspects of three atypical rhabdoid/teratoid tumors diagnosed in the Hospital A.B.C. of Mexico City. RESULTS: The three tumors were seen in boys aged 18 months, 5 years and 14 years, localized to the right cerebellum, left cerebral hemisphere and left cerebellum, respectively. Different markers were shown including: vimentin, cytokeratin, epithelial membrane antigen, gliofibrillare acid protein, synaptophysin, alpha fetoprotein, S100, (HHF-35) actin, cromogranin, neurofilaments, human chorionic gonadotrophin, placental alkaline phosphatase, desmin, CD99 and carcinoembryonary antigen. We established the morphological similarities and differences in our cases, and noted the profuse vascular proliferation which in one case resembled vascular neoplasia. We compared our results with other published cases. CONCLUSIONS: The atypical rhabdoid/teratoid tumor of the CNS may easily be confused with other primary or metastatic tumors since the rhabdoid characteristics do not only occur in this tumor. Also the rhabdoid aspect may be less marked and in some cases does not predominate. Some zones of the tumor may be identical to other embryonic tumors of the CNS and the immunophenotype be superimposed on that of other neoplasias of the nervous system. PMID- 11391488 TI - [Cerebellar hemorrhage in full-term newborn babies]. AB - INTRODUCTION: Cerebellar hemorrhage (CH) has been observed in 5 to 10% of the autopsies done on newborn babies. Since neuroimaging techniques have become available it is easier to diagnose the condition. In this paper we report on a series of cases of CH in full-term newborn babies. OBJECTIVES: To determine the number of patients with CH diagnosed by neuroimaging, make a descriptive study and analyze their progress. RESULTS: Between 1984 and 1999 six patients had CH, three boys and three girls, five born after their mother's first pregnancy. Four were vaginal births; in two forceps were used and in one a vacuum extractor; two were born by cesarean section. Four showed symptoms within the first 48 hours of life, one on the fourth day and one on the twenty fifth day. The latter had hemorrhagic disease of the newborn. In five patients transfontanellar ultrasound was useful in diagnosis. In all six cases computerized axial tomography scan confirmed the diagnosis. Cerebral magnetic resonance (MR) was done in three cases. No arteriovenous malformations were shown on angio-MR. Two patients had hydrocephalus and both were treated by ventriculo-peritoneal shunts. Three cases had transient ventricular dilatation which improved with medical treatment. The patient with hemorrhagic disease of the newborn had alterations in blood clotting. In three patients metabolic studies were normal. Five patients were treated conservatively and only one neurosurgically. Subsequent evolution was characterized by the presence of psychomotor retardation with mild cerebellar signs. At school age, only observed in two cases, there were learning difficulties with a low intellectual coefficient and problems with reading and writing. In one case there was epilepsy, controlled by use of two antiepileptic drugs. CONCLUSIONS: Half the cases of CH transfontanellar ultrasound is useful in diagnosis. Most patients were managed conservatively. During the clinical course there were psychomotor retardation, cerebellar signs, cognitive deficits with learning problems and epilepsy. PMID- 11391489 TI - [Etiology of late-onset epilepsy]. AB - INTRODUCTION: Late-onset epilepsy is considered the epilepsy that starts after 25 years old. PATIENTS AND METHODS: To determine the etiology and value of the clinical and paraclinical studies in the diagnosis, we evaluated 300 patients older than 25 years admitted in the Institute of Neurology and Neurosurgery of the city of La Havana between 1980 and 1990. Data about the illness, family and personal history and physical exploration were picked up. EEG, CT, MRI and carotid arteriography were performed. RESULTS: The main prevalence was 25 to 29 years (15.3%) and those older than 60 years (14.3%). We determined the diagnosis in 56% of the patients. Stroke was present in 19.3%, degenerative diseases in 16.6%, cerebral tumors in 9%, severe cranial trauma in 7.6% and infections, perinatal damage and toxic and metabolic causes in 3.3%. CONCLUSIONS: The symptomatic epilepsy is the most frequent type of late-onset epilepsy, the vascular and degenerative etiology are the most important in those older than 40 years old, these findings are similar to those of the developed countries. PMID- 11391490 TI - [Early onset adhalinopathy (LGMD2D) mimicking congenital muscular dystrophy]. AB - INTRODUCTION: The recent discovery of the dystrophin-associated complex of glycoproteins led to the delineation of sarcoglycanopathies, a phenotypically similar to dystrophinopathies group of clinically heterogeneous and progressive muscular dystrophies. The objective of this paper is to report the clinical, biochemical, histological, immunohistochemical and molecular genetics characteristics observed in a case of adhalinopathy (alpha-sarcoglycanopathy or LGMD2D) presenting in early months of life and resembling congenital muscular dystrophy. CLINICAL CASE: An 12-year old school boy, the third son of a healthy, young, non consanguineous couple, presented at birth with bilateral cleft lip, cleft palate and mild hypotonia. At age 6 months it was believed he suffered from a benign form of congenital muscular dystrophy on the basis of clinical, biochemical, electrophysiological and histological findings. From 5 years onwards he had frequent falls and climbing stairs had become increasingly difficult. Also, a positive Gowers 'sign, mild calf hypertrophy, high serum creatine phosphokinase level and myopathic electromyographic features were present; otherwise, cardiological evaluation and intelligence were normal. A repeated muscular biopsy at 10 years showed dystrophic features as well as selective deficiency of adhalin on immunostaining. DNA analysis demonstrated the patient being homozygote for a R77C mutation. Actually, a marked lumbar lordosis and waddling gait, an impossibility of climbing stairs and arising from the floor in addition to absent rotulian reflexes and mild Achilles retraction are present. CONCLUSIONS: LGMD2D may present in the first months of life mimicking congenital muscular dystrophy. It seems reasonable that biopsies of all new cases of muscular dystrophies be selectively immohistochemical analyzed, and when it is possible the diagnosis should be confirmed by DNA analysis. PMID- 11391491 TI - [Syringohydromyelia. Report of a case which resolved spontaneously]. AB - OBJECTIVE: To present a boy with syringohydromyelia associated with Chiari I malformation that showed spontaneous remission. CLINICAL CASE: A boy with moderate short stature and discrete dysmorphic facial features was studied by cerebral and spinal magnetic resonance (MR). He showed syringohydromyelic cavities in cervical and dorsolumbar regions associated with Chiari I malformation. MR study was performed every year and collapse of both syringohydromyelic cavities was found four years after discovering the picture. CONCLUSION: Spontaneous collapse of syringohydromyelia can be observed, though it occurs very seldom. PMID- 11391492 TI - [Gradenigo syndrome as the form of presentation of nasopharyngeal carcinoma]. AB - INTRODUCTION: Nasopharyngeal carcinoma is a condition which usually has an insidious onset and non-specific features in the initial stages, so it is difficult to make an early diagnosis. The most usual presenting features are otological (serous otitis media) and involvement of adjacent cranial nerves. We report a case of Gradeningo's syndrome due to the tumour spreading towards the base of the skull. We review the literature on the aetiology of this syndrome. CLINICAL CASE: A 53 year old patient required neurological assessment for a clinical condition which was compatible with Gradenigo's syndrome that involved both right V and VI cranial nerves. This study permitted diagnosis of a nasopharyngeal carcinoma which had been undetected because of its non-specific features, until this complication occurred. Otorhinolaryngological assessment proved the presence of a neoplasm in the cavum. Biopsy of the lesion showed it to be a well-differentiated squamous cell carcinoma. Cranial magnetic resonance imaging showed extension of the tumour to the base of the skull, adjacent to the right sinus cavernosus. The cerebrospinal fluid was normal. Treatment by radiotherapy was indicated. CONCLUSIONS: Diagnosis of nasopharyngeal carcinoma requires a high index of suspicion in view of its initial, sparse, non-specific symptoms. Although ideally the disease should be detected in its early stages, we believe that it is useful to recommend that in cases of Gradenigo's syndrome a full systematic otorhinolaryngological exploration be made so as to effectively rule out this disorder. PMID- 11391493 TI - [Sleep in infantile autism]. AB - INTRODUCTION: Analysis of nocturnal sleep in infantile autism has been presented in various studies. However, there has been no systematization including the different structural and paroxystic alterations at the same time as permitting the development of a general theory of the effect of sleep on prognosis and treatment, particularly in a spectrum in which there is currently no definite solution. DEVELOPMENT: A systematic review was made of the literature obtained from publications included in MEDLINE and web pages of the last 25 years using the key words: autism, Asperger's disorder, sleep, childhood and Rett's syndrome. Altogether 21 papers fulfllled criteria for inclusion. Disorders of sleep in infantile autism were classified into three types: immaturity of sleep, showing a destructured polysomnographic recording and negative correlation with the level of development; functional alterations of sleep with early waking and difficulty in going to sleep being the disorders most frequently seen; and paroxystic alterations with epileptiform discharges being the commonest, without necessarily occurring together with seizures. The opinions stated on questionnaires and the data observed on the polysomnography were not in agreement. CONCLUSIONS: Analysis of the literature has permitted us to make an initial classification of sleep disorders in autistic children, and has shown a marked presence of these disorders in the evolution of autistic children. It is necessary that further studies being done, polysomnographic rather than by means of questionnaires, for two reasons: clinical and in order to obtain more precise classification. PMID- 11391494 TI - [Intracranial lipomas]. AB - INTRODUCTION: This paper reviews current literature about intracranial lipomas with emphasis on histogenesis, diagnosis and therapy. DEVELOPMENT: Intracranial lipoma is a congenital malformation. Epilepsy, mental dysfunctions and headache are the most common clinical manifestations, being a third of the asymptomatic cases; certain localizations cause specific symptomatology of the involved area. Associate anomalies are given in half of the lipomas, being the most frequent the agenesia of the corpus callosus. The diagnosis is not possible based on the clinical data, so it is necessary the practice of complementary exams in every suspected cases. The neuroimaging studies (TC or MRI) usually permit the diagnosis of this entity in an objective way and practically of certainty. It has been adduced a numberless histogenic theory on the lipomas, being accepted the lipoma like congenital malformation at the present time. The surgery is contraindicated, however still play a role in the management of some cases of hydrocephalus and bony affectation. The medical control of the epilepsy is forced. CONCLUSIONS: The development of the neuroimaging techniques and the biggest compression in the histogenesis have increased the knowledge that one had of the intracranial lipomas, modifying their presage and giving up the aggressive surgery of last times. PMID- 11391495 TI - [Idiopathic generalized epileptic syndromes of children]. AB - INTRODUCTION: The idiopathic generalized epilepsies of children form a neurobiological continuum which starts during the first years of life, until adolescence. In this group, infantile absence epilepsy may be considered to be the maximum expression of the idiopathic generalized epilepsies in this age group. Infantile absences have seen studied in animal models, in humans and genetically. DEVELOPMENT: We analyze the clinical and encephalographic characteristics of infantile absence epilepsies together with their prognosis and epidemiology. We also consider the palpebral myoclonias with absences and absences with perioral myoclonias. The generalized tonic-clonic seizures of childhood form an ill-defined epileptic syndrome, with very heterogeneous clinical and electroencephalographic characteristics. CONCLUSION: It seem important to be able to establish the precise diagnosis in these syndromes, since correct treatment depends on it. PMID- 11391496 TI - [Neuropsychological assessment in posttraumatic amnesia disorder]. AB - INTRODUCTION: The neuropsychological evaluation of memory is an essential investigation in all patients with head injuries, since alterations occur in a high proportion of injured persons and are one of the commonest sequelae. OBJECTIVE: To analyse aspects of the neuropsychological evaluation of memory in head injuries, and the influence of other factors on memory. DEVELOPMENT: Regarding the study of memory capacity, other aspects relative to the presence or absence of posttraumatic amnesia (its characteristics and duration), lacunar-type disorders, retrograde amnesia, anterograde amnesia, ability to learn new information and the position of explicit and implicit memory should be considered. We analyze the instruments for measurement most commonly used for neuropsychological evaluation of memory in patients with head injuries, and the main batteries, tests and scales used for objective evaluation of the presence or absence of memory deficit. We review the main limitations and methodological problems presented by the tests and scales for measurement in this type of population. The instruments of measurement, except for those adapted for the detection of common alterations in head injuries often present serious problems when used to detect posttraumatic deficits. CONCLUSION: Clinically, evaluation of memory should show a profile of the patient's capacities, measure the changes in memory function over time, and predict memory function in everyday activities. PMID- 11391497 TI - [Evaluation of children's neuropsychological syndromes]. AB - INTRODUCTION: A new instrument of neuropsychological assessment, the Luria Neuropsychological Diagnostic Battery, has recently become available in Spain. This battery is based on a revision of the Luria Neuropsychological Diagnostic (Christensen, 1975) and has been standardized by Manga and Ramos (1991) to be used with children from 7 through 10 years old. DEVELOPMENT: This battery will be very useful for the neuropsychological evaluation of learning disabled and brain damaged children because profiles obtained based on it can be used to organize the treatment program based on the strengths and weaknesses of the child. A programme of psychological and pedagogical individualized intervention must start from a comprehensive evaluation, from an evaluation of the children as a whole person. The aim of our research is to produce several intervention (rehabilitation) principles for children with brain dysfunction, brain damaged and others clinical disorders, arose from neuropsychological assessment and from research works of psychological processes. We have realized clinical research with developmental dyslexic children, hyperactive children and epileptic children. PMID- 11391498 TI - [Neuropsychiatric disorders in Parkinson's disease]. AB - OBJECTIVE: This paper reviews the main neuropsychiatric disorders associated with Parkinson's disease (PD) and describes the neuropathological hypothesis proposed to explain these symptoms. DEVELOPMENT: This disease is usually associated with neuropsychiatric complications such as depression, anxiety and apathy. Besides, psychiatric symptoms are one of the most common side effects of antiparkinsonian drug-therapy. CONCLUSIONS: Depression is the most frequent emotional disorder reported in patients with PD. Up to 20% of parkinsonian patients meet DSM-IV criteria for major depressive episode and another 20% for dysthymia, while the prevalence of depression in normal aged population is about 2-8%. The relationship between PD and depression has not been fully established. Some investigators have suggested that depressive symptoms in PD are causally related to the underlying neuropathological process, affecting predominantly serotoninergic and dopaminergic pathways. Alternatively, depression in PD may represent a normal reaction to the progressive physical impairment induced by the disease. Otherwise, up to 20% of parkinsonian patients present levodopa-induced psychiatric complications. Visual hallucinations are the commonest, but delusions, confusional states, sexual disorders and sleep disorders have also been described. Serotonin and dopamine have been implicated in the neuropathological basis of these disorders. PMID- 11391499 TI - [Nosologic aspects of personality change due to head trauma]. AB - INTRODUCTION: Personality change due to head injury is one of the most prevalent neuropsychiatric posttraumatic disorders and causes significant impairment in familial, social or occupational functioning. OBJECTIVES: To study the prevalence and clinical characteristics of personality changes secondary to severe closed head injuries, according to DSM-IV criteria. PATIENTS AND METHODS: Fifty-five patients (aged 15-65 years) with severe head trauma were studied during the chronic stage (11-3 months) with the following instruments: the Revised Iowa Collateral Head Injury Interview for the assessment of posttraumatic frontal symptoms, the Standardized Polyvalent Psychiatric Interview of Lobo et al, assessment of premorbid personality and its exacerbation, and the Scale of Aggressiveness of Yudofsky. RESULTS: Thirty-three patients (60% of the sample) fulfilled DSM-IV criteria for personality change due to head injury; two thirds of them were combined or mixed type, which consists in the association of two or more types specified in the DSM-IV. The most prevalent types were apathic, unstable, disinhibited, aggressive, which are related with lesions in prefrontal cortex. There were nine patients with 'unspecified' symptoms, such as 'inappropriate euphoria' and 'poor insight', and other symptoms related to executive dysfunction. CONCLUSIONS: One third of cases of personality changes were related to premorbid features; this fact argues against the exclusion of the criterium 'excerbation of premorbid personality traits' in DSM-IV. Only one third of posttraumatic personality changes can be classified according to DSM-IV and ICD-10 criteria. The high prevalence of symptoms related to deficit in self awareness and executive dysfunction suggest the need of further investigations about nosology in this field. PMID- 11391500 TI - [Neurological anthropology among the Kamayura Indians of the Alto Xingu]. AB - INTRODUCTION: The Kamayura tribe is made up of 300 persons living in the Alto Xingu in Mato Grosso, Brazil. Their traditional system of health care is based on the paje, a witch doctor who uses plants and prayers for treatment. MATERIAL AND METHODS: Field work was done in the Kamayura village holding successive interviews with the chief and the paje Takuma to obtain information regarding the neurological disorders found there, the indigenous beliefs regarding illness (natural or based on witch-craft), their classification and traditional treatment based on plants. The terms employed were translated from Kamayura into Portuguese. RESULTS: Illness in Indians is caused by the revenge of the spirit (mama'e) of the animal killed by the huntsman. Epilepsy (Teawurup) or armadillo disease is caused when a huntsman kills an armadillo. It is treated with two roots, tsimo and wewuru, kneaded, diluted in water and applied to the eyes. An infusion of enamum root is also used. Migraine or monkey disease causes a pulsatile headache and vertigo. Mental retardation and infantile cerebral palsy are included in the ant-bear disease. Depression is treated with the plant 'iputunu'; which is applied diluted in water to the face of the patient so that he no longer sees his dead relations and may be cured. Schizophrenia or apuayat (owl disease) also occurs, but not parkinsonism or stroke. CONCLUSION: The Kamayura pajes have established a system of health-care based on magic folklore, transmitted orally and making use of traditional plants. PMID- 11391501 TI - [Neurologic complications in lymphoma patients]. PMID- 11391502 TI - [Vascular hereditary dementia CADASIL type in Colombia. III. Linkage analysis to notch3 gene]. AB - OBJECTIVE: To perform linkage analysis between the Short Tandem Repeats (STR) microsatellite markers D19S923, D19S929, D19S22, which are in strong genetic linkage to Notch3 gene in order to contrast the hypothesis that the vascular hereditary dementia phenotype described in a multigenerational extended pedigree from Colombia correspond to CADASIL (Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy). Even we know that using techniques as the Single Strand Conformational Polymorphisms (SSCP) could determine mutations in Notch3, the rationality of this approach is that intronic variations could not be defined and that we are interested in determine if some forms of the clinical presentation and its phenotypic variability make part of CADASIL. INTRODUCTION: The CADASIL phenotype is caused by mutations in the Notch3 gene. Clinical features of CADASIL are: 1. Recurrent cerebra-vascular episodes; 2. Migraine history; 3. History of transitory ischemic attack and, 4. Behavior changes and dementia. MATERIAL AND METHODS: By using SIMLINK we showed that the extended genealogy had the enough power to detect significant LOD (logarithm of oods) score values when Notch3 was considered the disorder cause. Linkage analysis was carried out by using parametric and non parametrical methods. The Elston-Stewart general method was used as the parametrical analysis and the sib pair method as the non-parametrical one. We perform simulations changing the affection status codification by including as affected or not including those individuals with migraine. Furthermore, in order to detect the stability of the results, we changed the penetrance values, the genetic frequencies on both, the marker loci and the affection locus. RESULTS: The maximum pair-wise LOD score was 2.04 which was detected at the marker D19S23 with q= 0.11cM. This distance correspond exactly with the Notch3 location. That is 100 times more probable that there is linkage that there is not. In other words this probability could be explained as if the phenotype correspond to CADASIL than to other vascular dementia. The non parametric results were compatibles with the parametric ones. When the migraine symptom was considered as a part of the affected status, the LOD score values showed not linkage. CONCLUSIONS: The results of the linkage analysis to these STR microsatellite markers suggest that the vascular hereditary dementia phenotype described in this family correspond to CADASIL caused by a polymorphism on the Notch3 gene. On the contrary, these same results suggest that the migraine phenotype is not a part of the progressive dementia. PMID- 11391503 TI - [Searching for more specific dimensions for the measurement of quality of life in multiple sclerosis]. AB - INTRODUCTION AND OBJECTIVES: Study of the quality of life in multiple sclerosis (MS) should consider use of an instrument which includes social, psychological, physical and mental aspects. The objective of this study is to show the validated dimensions of the scale of quality of life Functional Assessment of Multiple Sclerosis (FAMS) in the Spanish population, and verify their consistency. PATIENTS AND METHODS: 308 patients with MS (clinically defined or with laboratory support, according to Poser s clinical criteria), referred from 10 Neurology Outpatient Clinics of Hospitals of the Autonomic Community of Madrid. The study was of transversal design, and carried out by direct interview. We used a questionnaire containing the FAMS scale and other variables related to different aspects (clinical, social, employment, psychological and cognitive). RESULTS: The average age was 38.2 +/- 10.5 years with an average score on the Expanded Disability Status Scale (EDSS) of 3.6 +/- 2.3. The average scores of the parameters are lower than those obtained with the original version of FAMS; the internal consistency of these dimensions is slightly higher than those of the original FAMS. The parameters symptoms and general state of mind and also the FAMS in general had their internal consistency increased by the inclusion of eight variables which were originally excluded. CONCLUSION: The FAMS scale is more consistent and reliable for the evaluation of quality of life if we include specific variables of MS which do not form part of the American version of the FAMS. PMID- 11391505 TI - [Evaluation of motor and sensory neuroconduction of the median nerve in patients with carpal tunnel syndrome treated with non-coherent light emitted by gallium arsenic diodes]. AB - INTRODUCTION: The treatment selection in the carpal tunnel syndrome according to the damage of the median nerve is important and all of these have adverse effects. A good alternative without undesired reactions is irradiation of the carpal tunnel with not coherent light between 920 and 940 nm emitted by gallium arsenide diodes, resembling the physic and therapeutic laser effects. PATIENTS AND METHODS: Twenty-six female patients with idiopathic middle carpal tunnel syndrome were irradiated 15 minutes daily during three weeks. The median nerve motor and sensitive neuroconduction was studied before and immediately after the treatment. RESULTS: The abnormal neuroconduction variables (latency, amplitude and velocity conduction) did not modify when treatment concluded, in spite of all the patients reported disappearance of pain and numbness in damaged hands. CONCLUSIONS: Not coherent light does not change the fibers functional state explored by conventional neuroconductions techniques. It remains to know if this light produces fine fibers improvement. PMID- 11391504 TI - [Blood neuronal specific enolase in newborns with perinatal asphyxia]. AB - INTRODUCTION: Neuron-specific enolase (NSE) is a sensitive marker of brain injury after hypoxia or ischemia. There are few studies about its usefulness in asphyctic newborns. OBJECTIVE: To study the correlation between blood NSE levels and neurological outcome in newborns with hypoxic ischemic encephalopathy. PATIENTS AND METHODS: We have determined the blood values of NSE by radioimmunoassay in 25 asphyctic term-newborns with clinical encephalopathy (of mild, moderate and severe degree) and in 22 healthy term newborns (control group). Blood samples were obtained between 24 and 72 hours after birth in all neonates. Surviving infants were followed for a variable time (median: 3.5 years; range: 1-6) and the neurological status was determined. RESULTS: NSE levels in the group of asphyctic neonates who died or developed neurological sequelae (n= 6; mean: 116.4 ng/ml; range: 42-226) were significantly higher than NSE values in the group of asphyctic neonates who were neurologically normal at follow-up (n= 19; mean: 21.3 ng/ml; range: 7.4-40), with p< 0.001. There were not differences between NSE values in the group of asphyctic neonates who developed neurologically normal and the control group (mean: 7.6 ng/ml; range: 10.3-28.3). Sensitivity and specificity of blood NSE as predictor of poor outcome were, respectively, 100% and 78%. The combined specificity for blood NSE together with a moderate/severe encephalopathy was 95%. CONCLUSIONS: The presence of elevated NSE values in blood after perinatal asphyxia can be a sensitive indicator of conspicuous brain damage. The combined information provided by the severity of the encephalopathy together with the blood NSE values have shown a high predictive value for neurological outcome. PMID- 11391506 TI - [Behr's syndrome. a report of seven cases]. AB - OBJECTIVE: To present the clinical and radiological findings of a large series of cases of Behr's syndrome. PATIENTS AND METHODS: We studied 7 cases, 6 of whom were 3 pairs of siblings, with clinical alterations compatible with Behr's syndrome which were studied from clinical, genetic and biochemical points of view. In 5 cases magnetic resonance (MR) imaging was used. RESULTS: All patients had marked pyramidal signs, pallor, optic atrophy and normal laboratory findings in the biochemical investigations done to rule out metabolic disorders. MR showed atrophy of the cerebellum which was marked in 3 cases and moderate in 2. In the other two cases it was not used, since it was not available when those cases were studied. CONCLUSIONS: Behr's syndrome is not usually considered when studying a patient with ataxia associated with other pyramidal and ocular alterations, due to the theoretically diverse origins of the condition which is a syndrome, not a disease. Perhaps the presence of frank atrophy of the cerebellum may differentiate it from many other clinically similar disorders, which do not however show true cerebellar ataxia. PMID- 11391507 TI - [The quality of life of patients with strokes: from the point of view of factors which may affect it]. AB - OBJECTIVE: To evaluate the effect of clinical and psychosocial factors on the quality of life of patients with stroke. PATIENTS AND METHODS: A non-experimental transverse study was done on 39 patients with strokes seen in the cerebrovascular diseases clinic of the Institute de Neurologia y Neurocirugia in La Havana (Cuba). We recorded data on clinical variables (age, sex, race, previous illnesses, type of stroke, vascular territory, classification of prognosis, etiopathogenesis and function on the Rankin scale) and on psychosocial variables (pattern of behavior, anxiety as a feature and state, depression, manner of coping and social support received). Correlations were established with the dependent variable Quality of life, evaluated on the Nottingham Health Profile. RESULTS: Of the clinical variables there was no correlation with age, female sex was related to greater isolation and the presence of diabetes mellitus with less energy. Greater isolation was seen to be correlated with increased score on the Rankin scale, but this association was not statistically significant. Similarly the clinical characteristic of anxiety was related to greater difficulty in sleeping, as also occurred in depression. The latter was also associated with less energy and greater social isolation. The characteristics of self-control, acceptance of responsibility and positive reevaluation had a positive relation with energy and physical mobility. The greater the social support, especially in the area of health, the better the quality of life in the realms of pain, sleep and social isolation. CONCLUSION: The psychosocial aspects had more influence on the quality of life of these patients than the clinical factors did. PMID- 11391508 TI - [Partial non convulsive epileptic status as initial presentation of limbic encephalitis]. AB - INTRODUCTION: Limbic encephalitis is an unusual presentation of paraneoplastic syndrome, which includes among its symptoms seizures. CLINICAL CASE: We report a case with a rare presentation of limbic encephalitis as initial symptom of small cell lung carcinoma. A 69 year-old woman presented with partial non convulsive status epilepticus and neuropsychiatric disturbances. Chest radiography and computed tomography showed mediastinal lymphadenopathy and lung nodules. Subsequently, small cell lung carcinoma was diagnosed by lymph node biopsy. The cerebrospinal fluid study was normal. The electroencephalography and magnetic resonance imaging (MRI) findings had distinctive features compatible with temporo limbic dysfunction. The anti-Hu antibodies were negative. The neuropsychiatric symptoms improved significantly after systemic chemotherapy and adjuvant radiotherapy. A serial follow-up MRI of the head showed no evidence of intracranial metastasis three months after the diagnosis of cancer. Limbic encephalitis may be an initial manifestation of lung cancer. Paraneoplastic limbic encephalitis is considered a remote effect of cancer commonly associated with anti-neuronal antibodies (anti-Hu) and small cell lung carcinoma. CONCLUSIONS: Status epilepticus could be an early sign of limbic encephalitis. The absence of anti-Hu antibodies does not rule out the presence of an underlying small cell lung carcinoma in patients with a clinical diagnosis of limbic encephalitis. Greater awareness for diagnosis and early treatment of the primary tumor offers the best chance for improvement in patients with lung cancer presenting with limbic encephalitis. PMID- 11391509 TI - [Focal cortical dysplasia and refractory epilepsy. Surgical treatment]. AB - INTRODUCTION: Focal cortical dysplasia (FCD) is an unusual cause of refractory epilepsy, in which the morbid anatomy is characterized by cortical laminar dysplasia and the presence of balloon like cells. CLINICAL CASE: A 36 year old woman who had had drug-resistant epilepsy since the age of 9 years old, with daily complex partial seizures and seizures in which she fell to the ground. After many therapeutic trials, at the age of 29 years she had a callostomy as palliative treatment for the seizures in which she fell and was completely cured of these. The complex partial seizures increased in frequency, in spite of high doses of multiple drugs, so the possibility of resective surgery was considered. A surface EEG with predominant lateralization and localization to the left temporal lobe, ictal and interictal cerebral SPECT, PET and MR were done. The findings were comparable with two foci of dysplasia in the left cerebral hemisphere, one superior parietal and the other posterior basal temporal in situation. Finally, video-EEG monitoring with foramen ovale electrodes showed ictal activity starting in the left posterior temporal regions and with rapid homolateral anteromesial diffusion. A left temporal lobectomy and amigdalohippocampectomy was done with resection of a posterior basal temporal lesion. This reduced the number of seizures by 90%, with minimal dysnomy as a sequel. The findings on morbid anatomical study were compatible with a FCD. CONCLUSIONS: FCD is a cause of refractory epilepsy which may benefit from surgical treatment, with excellent results, after suitable pre-operative surgical evaluation and planning, including hippocampal evaluation since there is a high incidence of associated mesial sclerosis. PMID- 11391510 TI - [Microvascular decompression of trigeminal neuralgia caused by vertebrobasilar dolichoectasia]. AB - INTRODUCTION: Trigeminal neuralgia due to vertebrobasilar dolichoectasia is an acquired disease whose true incidence is not known. Microvascular decompression is the most effective technique both for symptomatic relief and for the conservation of nerve structure and function, in spite of the potential complications of all major surgery. In cases which are drug-resistant and have a life expectancy of over five years, microvascular decompression may be done using several techniques. CLINICAL CASES: We present three cases with drug-resistant trigeminal neuralgia. One patient had a history of previous cerebrovascular ischaemia, another had arterial and pulmonary hypertension. In all cases vertebrobasilar dolichoectasia was seen on magnetic resonance. Microvascular decompression of the trigeminal nerve was done, placing pieces of Teflon between the tentorium and the artery to displace it. Postoperatively the pain disappeared in two cases and was much relieved in the other, making good progress with no complications. CONCLUSIONS: Trigeminal neuralgia due to vertebrobasilar dolichoectasia is a progressive acquired disease. The imaging technique of choice is magnetic resonance. Cerebral angiography may be useful for confirmation. This new technique of microvascular decompression avoids excessive manipulation of the ectatic, arteriosclerotic basilar artery and also pulsatile compression, so that the risk of secondary effects is reduced and the efficacy of the decompression maintained. Further cases are needed to confirm the usefulness of this technique. PMID- 11391511 TI - [Ring chromosome 8: microcephaly, mental retardation and minor facial anomalies with adhesive behavioral phenotype]. AB - INTRODUCTION: Since the first description of ring chromosome 8 [r(8)] in 1973, only a few patients have been reported. In this report we present a child with this anomaly, and we compare his clinical manifestations with previously reported patients. CLINICAL CASE: A 12 year-old boy presented with low birth weight, mental retardation, microcephaly, short stature, hypotonia, and minor facial anomalies: hypotelorism, bilateral epicanthic folds, long philtrum, thin lips, narrow palate, micrognathia and low-set ears. Digital anomalies were bilateral brachyclinodactily of the fifth finger, and cutaneous syndactyly between second and third fingers. The boy had a pleasant personality but exhibit attachment for people and things with unrestricted affect. Cytogenetic analysis on peripheral white blood cells showed a r(8) chromosome. Parental karyotypes were both normal. CONCLUSIONS: The major features in the boy studied by us were found also in the other cases reported with an r(8) chromosome, but all they were non-specific features, and do not support the existence of a readily recognizable r(8) chromosome syndrome . Follow-up data with special emphasis on the behavioral characteristics are needed for defining an specific behavioral phenotype. PMID- 11391512 TI - [The CADASIL syndrome: a model of subcortical-cortical disconnection]. AB - INTRODUCTION: CADASIL syndrome (Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarts and Leukoencephalopathy) includes some neurological signs and symptoms (gait disturbances, epileptic seizures, pseudobulbar palsy, migraines, etc.), as well as neuropsychological dysfunctions (cognitive and executive impairment, emotional disorders and, frequently, dementia). This syndrome is a good model of white matter damage and potential disconnection syndromes. CLINICAL CASE: In this article, the neuropsychological profile of a 47 year-old woman diagnosed of CADASIL is investigated and thoroughly discussed. Results show the presence of a moderate cognitive and executive impairment, specially of memory, psychomotor abilities, executive functions and verbal fluency, but not dementia, overall suggesting the presence of a temporal-frontal-subcortical disfunction. CONCLUSION: This clinical pattern is an illuminating example of the neuropsychological consequences of the partial disconnection of prefrontal cortex from the thalamus and basal ganglia. PMID- 11391513 TI - [The physiology of the lid motor system]. AB - OBJECTIVE: The physiology and anatomy of the lid motor system have been widely studied from experimental and clinical points of view during last years. These studies, mainly carried out in animals, have helped to understand its functioning in normal and pathologic situations. Despite the apparent simplicity of this motor system, lids are able of doing many types of movements, depending on each particular situation. Besides participating in reflex and spontaneous blinks, lids take part actively in motor responses of major complexity and that requires a better precision, for instance, during emotional responses as laughing or crying in human beings. Lid movements also accompanies the eyes in such a manner not to cover the pupil while they maintain a fixed position in the orbit or during reflex and voluntary saccades. DEVELOPMENT: This paper deals with the neural circuits underlying lid motor system and how the activity in them translates, through the corresponding muscles, in a coherent response despite the different situations imposed by the different stimuli. In this way, it is described each element and how they contribute in the whole we observe as lid movements, in its repertoire. CONCLUSION: The facial motor system has been quite preserved through phylogeny, so it can be said that human beings possess a mechanism for lubricating and protecting the eye that, except for certain differences, share with the rest of the terrestrial living beings. PMID- 11391514 TI - [Huntington's disease: a bimolecular vision]. AB - INTRODUCTION: Huntington s disease is a genetic autosomal dominant progressive neurodegenerative disorder determined by mutation at the gene that codes for the protein huntingtin, whose function is unknown. Clinically hallmarked by chorea and behavioral disturbances, the diagnosis is confirmed by blood test for the disease s gene. Experimental models of the disease, and new tools for in vivo and in vitro investigation are contributing for understanding its pathophysiology. DEVELOPMENT: The neurodegeneration is accomplished by apoptosis and predominantly strikes the striatum. Multiple evidence have emerged that oxidative stress, promoted by excess glutamate stimuli and iron deposits in the striatum play an important role, besides mitochondrial oxidative disfunction and reduced blood cerebral perfusion. There is no curative therapy for Huntington s disease. Current treatment usually includes a neuroleptic for chorea and behavioral disturbances, and a serotonin-selective reuptake inhibitor when depression is present. There is a great hope that new knowledge about the pathophysiology of Huntington's disease will engage in a better treatment, but neurotransplantation is an alternative treatment under development. PMID- 11391515 TI - [Photosensitive epilepsies of children]. AB - INTRODUCTION: Reflex epileptic seizures are caused by a specific sensorial stimulus which determines their classification. Photosensitive epilepsies are the commonest forms and are included with the idiopathic generalized epilepsies. DEVELOPMENT AND CONCLUSIONS: We analyze the different responses to intermittent light stimulation, in both normal and epileptic persons, and study the various epileptic syndromes in which photosensitivity is seen. The purely photosensitive (photogenic) epilepsies are characterized by seizures caused only by light as compared with the epilepsies with photosensitivity which also present spontaneous seizures. Special mention is made of seizures induced by the television screen, computer screen and video-games. The palpebral myoclonias with absences may be considered to be reflex seizures, since they are induced by eyelid closure, and include the vary rare self-induced epileptic seizures. Finally we study the epileptic seizures induced by pattern and exclusively due to intermittent light stimulation. PMID- 11391516 TI - [The detection of possible cases of simulation after a traumatic brain injury]. AB - OBJECTIVE: To show the degree of detection of simulation, defined as the conscious, intentional production of false or exaggerated physical and psychological symptoms, motivated by external rewards such as payments by insurance companies and compensations. DEVELOPMENT: In forensic circles, a major problem is under consideration, namely recognition of the existence and nature of cognitive alterations after mild head injury (HI), since it is estimated that in 5-10% of the cases of mild HI there may be simulation of cognitive and emotional deficit, with further difficulty of differential diagnosis between simulation and the post-concussion syndrome. CONCLUSIONS: The forensic assessment of cognitive alterations following HI should include a clinical interview and neuropsychological evaluation. The former helps to determine the causal relationship between the traumatic accident and the resulting damage, continuance of symptoms and the existence or not of a premorbid pathological state. Although there is still not completely reliable, valid marker which permits one to be completely certain that the person is a simulator, neuropsychological evaluation permits suspicion of it in three aspects: 1. General indicators showing lack of internal consistence in the results of the person evaluated. 2. Specific tests to detect possible cases of simulation. 3. Profiles of responses which characterize possible simulators and neuropsychological evaluation tests. PMID- 11391517 TI - [Forensic neuropsychology in the aging and the dementias]. AB - INTRODUCTION. Forensic Neuropsychology establishes an expert valuation of the brain-injured patient (or supposed), or of the law offender people (or supposed) requiring law protection due to their illness. This discipline has a fundamental role in the study of cerebral organic syndromes, particularly in the dementias. DEVELOPMENT. The demential syndrome is common to a group of diseases as degenerative or cardiovascular disorders, brain tumors, syphilis, alcoholism or toxic factors, etc. When speaking of dementia we refer to a syndrome characterized by a progressive and global deterioration of the cognitive functions (memory, language, attention, space-temporal orientation, praxis, thinking, etc.) with preservation of the level of conscience (DSM-IV). This symptomatology affects the personality of the individual as reasonable being , as well as to his behavior and social adaptation. The decrease or loss of the intellectual and volitives abilities of the affected person of a demential syndrome, if it is permanent, also implies a change in its legal situation, since its legal capacity is altered. Therefore, it will be necessary to adopt protective measures for his person and his patrimony. Even if it is necessary, to promote a process of disability, whose sentence will be emitted by a judge, who will indicate: the degree of the mentioned disability, the trusteeship regime and who is designated as a legal tutor. PMID- 11391518 TI - [Forensic neuropsychology: principal issues and applications]. AB - INTRODUCTION: The term Forensic Neuropsychology refers to the application of the knowledge of the field of Neuropsychology to the legal matters. The field of Clinical Neuropsychology has experienced a great development, thanks to the investigations and publications that have been made both in the basic and applied fields. Nevertheless, only in the last years, clinical neuropsychologists have participated actively in the courts. DEVELOPMENT: The testimony as experts of neuropsychologists is more and more common. They inform to the judge on the behavioral, emotional and cognitive sequels following lesions to the human brain, as well as the importance of psychosocial variables in modulating brain function and dysfunction. The relationship between Neuropsychology and Law has been influenced by the confluence of a series of related factors that have contributed to the professionalization of this discipline. Among these, we can emphasize the study of brain-behavior relationships by means of quantitative methods, the interpretative strategies to infer the presence, location and type of neuropathology; the decision making on legal subjects like the determination of disability; the description of the characteristic cognitive-behavioral profiles of neurological disorders; the implication of the psychosocial variables and the decision on the most appropriate interventions. Some of the most important objectives of this new discipline are: the identification of the sequels of mild traumatic brain injury as the posconmocional syndrome , the determination of premorbid intellectual functioning and the valoration of cognitive loss. There is no doubt that in the next years this field of application of Clinical Neuropsychology is going to experience an important growth in our country, thanks partly, to the norms that are followed to establish the degree of disability and the compensations of the insurance agencies, that consider these neuropsychological sequelae. PMID- 11391519 TI - [The neuroanatomy of Juan Valverde de Amusco and medicine at the time of the Spanish renaissance]. AB - Juan Valverde de Amusco (c. 1525-c. 1564) is considered to have been the most important Spanish anatomist of the XVI century. A follower of Vesalius, he increased and divulged knowledge of anatomy during the Renaissance and his book The history of the composition of the human body was printed in Rome in 1556. The objective of this paper is to study the neuroanatomy in this book and present unpublished biographical data and describe the main contributions of this Castilian doctor to the neurosciences, in the context of Spanish medicine during the Renaissance period. He was born in the town of Hamusco (today Amusco) in the province of Palencia, which belonged to the Crown of Castile. Juan Valverde emigrated to Italy to improve his scientific knowledge. He carried out anatomical studies using the then revolutionary method of direct observation, as opposed to the Galenic criteria of authority inherited from the Medieval period. He trained in Padua under Realdo Colombo and lived in Rome where he practiced medicine until his death, becoming deservedly famous. He did not return to Spain since in the Spanish universities of the time there was a mentality which was reactionary to modern anatomy. His works, published in Italy but in the Spanish language, give an idea of the power of the Crown of Castile in the Europe of that period. The book is profusely illustrated with the first illustrations ever published in the history of printing, drawn by Nicolas Beatrizet. The book was sold widely and was translated and reedited on many occasions, until well into the XVIII century. For the first time Valverde made precise references to the minor circulation. He was the first anatomist to describe the muscles for movement of the eye correctly and the intracranial course of the carotid arteries. In his work he made the first drawing of the stapes, described by the Valencian Luis Collado. Vesalius and Valverde contributed decisively to the beginnings of modern neuroanatomy. Thanks to them, the brain is no longer an organ unknown to science. PMID- 11391521 TI - Liver cell volume regulation: size matters. PMID- 11391522 TI - Increasing incidence and mortality of primary intrahepatic cholangiocarcinoma in the United States. AB - Clinical observations suggest a recent increase in intrahepatic biliary tract malignancies. Thus, our aim was to determine recent trends in the epidemiology of intrahepatic cholangiocarcinoma in the United States. Reported data from the Surveillance, Epidemiology, and End Results (SEER) program and the United States Vital Statistics databases were analyzed to determine the incidence, mortality, and survival rates of primary intrahepatic cholangiocarcinoma. Between 1973 and 1997, the incidence and mortality rates from intrahepatic cholangiocarcinoma markedly increased, with an estimated annual percent change (EAPC) of 9.11% (95% CI, 7.46 to 10.78) and 9.44% (95%, CI 8.46 to 10.41), respectively. The age adjusted mortality rate per 100,000 persons for whites increased from 0.14 for the period 1975-1979 to 0.65 for the period 1993-1997, and that for blacks increased from 0.15 to 0.58 over the same period. The increase in mortality was similar across all age groups above age 45. The relative 1- and 2-year survival rates following diagnosis from 1989 to 1996 were 24.5% and 12.8%, respectively. In conclusion, there has been a marked increase in the incidence and mortality from intrahepatic cholangiocarcinoma in the United States in recent years. This tumor continues to be associated with a poor prognosis. PMID- 11391523 TI - Steatosis accelerates the progression of liver damage of chronic hepatitis C patients and correlates with specific HCV genotype and visceral obesity. AB - The role of steatosis in the progression of liver damage in chronic hepatitis C (CHC) was studied. Enrolled were 180 consecutive liver biopsy-proven CHC patients and 41 additional subjects with a known duration of infection. We evaluated the histological activity index (HAI), grade of fibrosis and steatosis, body mass index (BMI; kg/m(2)), distribution of body fat, HCV genotype, and levels of HCV RNA. Eighty six (48%) patients showed steatosis, and a higher prevalence was observed in genotype 3a infection (P <.01). A correlation between the grade of steatosis and fibrosis was observed (P <.001). Fibrosis was also associated with age (P <.001). After adjusting for age, the association between steatosis and fibrosis remained significant. The grade of steatosis also correlated with the HAI (P <.007) with a significant increase in periportal necrosis. No relation was found between steatosis and age, gender, iron storage, or levels of HCV RNA. Patients with a high grade of steatosis (>30%) showed higher serum levels of gamma-GT and ALT (P <.001). Overall, steatosis was not significantly associated to BMI. Analysis by single genotype showed a significant association between the grade of steatosis and BMI in type 1 infection r =.689; P <.001) and with levels of HCV RNA in type 3a infection r =.786; P <.001). Visceral fat distribution rather than BMI proved to be associated with steatosis (P <.001). Data obtained from patients with a known date of infection confirmed that steatosis grades 3-4 were associated with a higher annual rate of fibrosis progression, and showed that alcohol and steatosis act together in increasing fibrosis (P <.05). Our data indicate that steatosis is an important cofactor in increasing liver necroinflammatory activity and in accelerating fibrosis in CHC. Visceral obesity and genotype 3a play a role in the development of steatosis. PMID- 11391524 TI - Partially reversible renal tubular damage in patients with obstructive jaundice. AB - Decreased serum uric acid levels resulting from renal urate wasting have been occasionally encountered in jaundiced patients. However, in these cases, there are no data concerning the underlying renal tubular defects. In the present study, we investigated the renal tubular function in 35 patients with obstructive jaundice of various severity and causes (11 with lithiasis, 17 with carcinoma, and 7 with intrahepatic cholestasis). A detailed study of the renal tubular function was performed. Beyond the conventional methods, (1)H-NMR spectroscopy of urine was used to evaluate noninvasively renal damage by the characteristic perturbation in the excretion pattern of low-molecular weight endogenous metabolites. On admission, patients with obstructive jaundice had significantly lower serum uric acid and phosphate levels and higher bile acid concentrations compared with 40 age- and sex-matched controls. Serum uric acid levels presented a negative correlation with the total and direct bilirubin as well as the fractional excretion of uric acid. Furthermore, a great number of the patients studied developed one or more proximal tubular dysfunction manifestations beyond uricosuria, such as renal glucosuria, phosphaturia, and increased excretion of alpha(1)-microglobulin. (1)H-NMR spectroscopy of the urine showed decreased levels of citrate and hippurate and increased levels of 3-hydroxybutyrate and acetate. In 12 patients partial or complete remission of jaundice was followed by an improvement of the proximal renal tubular damage. In conclusion, obstructive jaundice can cause a partially reversible generalized proximal tubular dysfunction. PMID- 11391525 TI - Increased density of brain histamine H(1) receptors in rats with portacaval anastomosis and in cirrhotic patients with chronic hepatic encephalopathy. AB - The binding properties and the regional densities of histamine H(1) receptors were studied in brain of rats with portacaval anastomosis (PCA) and in autopsied brain tissue from cirrhotic patients with hepatic encephalopathy (HE). Receptor binding studies and quantitative receptor autoradiography were performed, employing [(3)H]mepyramine. Histamine H(1) receptors in rat brain displayed a higher density and a lower affinity compared with control human frontal cortex. Specific [(3)H]mepyramine binding was heterogeneously distributed throughout the brain in both species. In human brain, binding was highest in the parietal and temporal cortices and lowest in caudate-putamen. In rat brain, binding was highest in the suprachiasmatic nucleus, dentate gyrus of the hippocampus, ventromedial hypothalamus, and nucleus accumbens. Cortical tissue from PCA rats and frontal cortical tissue from HE patients contained significantly increased densities (B(max)) of H(1) receptors. A selective increase in H(1) receptor density was also observed in parietal and insular cortices of HE patients. Results of the present study suggest a selective up-regulation of brain H(1) receptors in PCA rats and in patients with HE. The central histaminergic system is implicated in the control of arousal and circadian rhythmicity. Previous studies have shown that blockade of H(1) receptors in PCA rats results in improved locomotor activity and circadian rhythmicity scores. The present findings suggest that cortical histaminergic hyperactivity could contribute to the neuropsychiatric symptoms characteristic of human HE, and that selective histamine H(1) receptor antagonists could be beneficial in the prevention and treatment of some of the symptoms of HE in cirrhotic patients. PMID- 11391526 TI - Global changes in interleukin-6-dependent gene expression patterns in mouse livers after partial hepatectomy. AB - Liver regeneration following 70% partial hepatectomy leads to rapid activation of genes in the remnant liver. Interleukin-6 deficient (IL-6 -/-) mice have impaired liver regeneration and abnormalities in immediate early gene expression. In this study, the gene expression program in the IL-6 +/+ and -/- livers at 2 hours posthepatectomy was examined with a cDNA array representing 588 highly regulated mouse genes. Thirty-six percent of the 103 immediate early genes were induced differently in IL-6 +/+ compared with IL-6 -/- livers, implying regulation by IL 6. IL-6 treatment of the IL-6 -/- mice in the absence of hepatectomy induced a much smaller set of genes in the liver, suggesting that IL-6 cooperates with other hepatectomy-induced factors to activate the large number of genes. Northern blot analyses were used to verify gene expression data obtained from the arrays. The expression of urokinase type plasminogen activator receptor (uPAR) and plasminogen activator inhibitor-1 (PAI-1), critical components of the urokinase plasminogen activator (uPA) system, was lower and delayed in IL-6 -/- livers. Despite the fact that active uPAR/uPA complex is critical for hepatocyte growth factor (HGF) activation, no differences were detected between the IL-6 +/+ and -/ livers in HGF activation as measured by receptor phosphorylation. On the contrary, the mitogen-activated protein kinase (MAPK) pathway was activated in IL 6 +/+ livers early during regeneration but remarkably delayed in IL-6 -/- livers. Defective liver regeneration may be explained by the large number of gene activation pathways altered in IL-6 -/- livers and further supports the finding that IL-6 is necessary for normal liver regeneration. PMID- 11391527 TI - Expression of anti-OV6 antibody and anti-N-CAM antibody along the biliary line of normal and diseased human livers. AB - Following hepatic injury, proliferation of anastomosing ductules can be observed. The origin of this ductular reaction is not completely clear, although there is considerable evidence for proliferation of a putative hepatic progenitor cell, reported to be located in the canals of Hering (CoH) and showing morphologic similarities with rat oval cells. In this study, we analyzed the immunophenotype of solitary oval cell-like cells (SOC), intralobular groups of cuboidal cells that might represent lining cells of CoH, bile ductular cells (BDC), bile duct epithelial cells (BEC), and hepatocytes. We used the antibodies OV6, CK19, and CD56 (NCAM) in a double-staining method in a series of 111 liver specimens. The series consisted of a variety of liver diseases, primary liver tumors, and normal livers. In normal livers, SOC, CoH, BDC, and BEC were uniformly and predominantly CK19+, OV6+, and CD56-. In diseased livers SOC and BDC were CK19+, OV6+, and also CD56+. Occasionally, BEC was CD56+ in damaged bile ducts in diseased liver, e.g., PSC. CoH lining cells were not present in cirrhotic nodules and were indistinguishable from BDC in the fibrous septa. The consistent and uniform staining patterns of SOC, CoH, and BDC support the concept that these cells share the same biliary lineage and might represent one biliary structure. The expression of CD56 on these cells in diseased livers indicates that CD56 is a useful marker for a reparative or regenerative state of the biliary liver-cell constituents but not to discriminate a putative hepatic stem cell. PMID- 11391528 TI - Liver transplantation for hepatocellular carcinoma: expansion of the tumor size limits does not adversely impact survival. AB - The precise staging of hepatocellular carcinoma (HCC) based on the size and number of lesions that predict recurrence after orthotopic liver transplantation (OLT) has not been clearly established. We therefore analyzed the outcome of 70 consecutive patients with cirrhosis and HCC who underwent OLT over a 12-year period at our institution. Pathologic tumor staging of the explanted liver was based on the American Tumor Study Group modified Tumor-Node-Metastases (TNM) Staging Classification. Tumor recurrence occurred in 11.4% of patients after OLT. The Kaplan-Meier survival rates at 1 and 5 years were 91.3% and 72.4%, respectively, for patients with pT1 or pT2 HCC; and 82.4% and 74.1%, respectively, for pT3 tumors (P =.87). Patients with pT4 tumors, however, had a significantly worse 1-year survival of 33.3% (P =.0001). An alpha-fetoprotein (AFP) level > 1,000 ng/mL, total tumor diameter > 8 cm, age > or = 55 years and poorly differentiated histologic grade were also significant predictors for reduced survival in univariate analysis. Only pT4 stage and total tumor diameter remained statistically significant in multivariate analysis. Patients with HCC meeting the following criteria: solitary tumor < or = 6.5 cm, or < or = 3 nodules with the largest lesion < or = 4.5 cm and total tumor diameter < or = 8 cm, had survival rates of 90% and 75.2%, at 1 and 5 years, respectively, after OLT versus a 50% 1-year survival for patients with tumors exceeding these limits (P =.0005). We conclude that the current criteria for OLT based on tumor size may be modestly expanded while still preserving excellent survival after OLT. PMID- 11391529 TI - Earlier expression of the transcription factor HFH-11B diminishes induction of p21(CIP1/WAF1) levels and accelerates mouse hepatocyte entry into S-phase following carbon tetrachloride liver injury. AB - Partial hepatectomy (PH) or toxic liver injury induces the proliferation of terminally differentiated hepatic cells to regenerate the original size of the adult liver. Previous PH liver regeneration studies showed that premature transgenic expression of the Forkhead Box M1b (FoxM1b, HFH-11B) transcription factor accelerated hepatocyte entry into DNA replication (S-phase). In this study, we used carbon tetrachloride (CCl(4)) liver injury to induce a different type of mouse liver regeneration and show that premature hepatic HFH-11B levels also accelerate the onset of hepatocyte S-phase in this injury model. Unlike PH liver regeneration, earlier hepatocyte proliferation after CCl(4) liver injury is correlated with diminished transgenic hepatic levels of p21(CIP1/WAF1) at the G1/S transition of the cell cycle. Differential hybridization of cDNA arrays and RNase protection studies determined that CCl(4) regenerating liver of transgenic mice displayed early stimulated expression of the S-phase promoting cyclin D1 and cyclin E and sustained levels of Cdc25a phosphatase genes. Compared with previous PH liver regeneration studies, our data suggest that premature expression of HFH 11B activates distinct S-phase promotion pathways in the CCl(4) liver injury model. Although proliferating transgenic hepatocytes induced by either PH or CCl(4) liver injury displayed early expression of identical M-phase cyclin genes (cyclin B1, B2, A2, and F), only CCl(4) regenerating transgenic liver exhibited earlier expression of the M-phase promoting Cdc25b. These studies suggest that CCl(4) injury of transgenic liver not only uses the same mechanisms as PH to mediate accelerated hepatocyte entry into mitosis, but also promotes M-phase entry by stimulating Cdc25b expression. PMID- 11391530 TI - An early lesion in hepatic carcinogenesis: loss of heterozygosity in human cirrhotic livers and dysplastic nodules at the 1p36-p34 region. AB - Loss of heterozygosity (LOH) of chromosome 1 has been suggested, by karyotyping, to be an initial episode in human hepatocarcinogenesis. However, this alteration has not yet been investigated in cirrhotic nodules (CNs) or dysplastic nodules (DNs). In an initial study from explanted or resected cirrhotic livers, LOH in 1p36-p32 was examined in 31 hepatocellular carcinomas (HCCs), 25 low-grade dysplastic nodules (LGDNs), and 24 high-grade dysplastic nodules (HGDNs). In HCCs, LOH was detected most frequently at loci D1S2843 (1p36.1) (28.6%), D1S513 (1p34.3) (29.2%), and MYCL1 (1p34.1) (28.6%). In HGDN and LGDN, LOH incidences at D1S513 were 11.1% and 13.6%, respectively. To further refine those results and to determine sequential relationships among CN, DN, and HCC, LOH was next studied in an additional 53 HCCs, 56 HGDNs, 30 LGDNs, and 215 CNs from 11 explanted human cirrhotic livers, including 30 "nodule-in-nodule" lesions. Seven markers between D1S2843 (1p36.12) and MYCL1 (1p34.1), and 1 each at D1S484 (1q24.1), IGF2R-3 (6q26), and TP53 (17p13.1) were used. LOH at D1S2843 and D1S513 was detected in HCCs (20.4% and 23.5%, respectively), HGDNs (7.7% and 18.5%), LGDNs (13.6% and 6.9%), and CNs surrounding either HCCs or DNs (7.4% and 8.3%). These results demonstrate that LOH at D1S2843 and D1S513 are early events in human liver carcinogenesis. Data from CN surrounding either HCCs or DN, and also nodule-in nodule lesions, provide evidence supporting a CN-->DN-->HCC progression. Different deletion patterns from multiple HCCs and DNs suggest independent origins for carcinogenesis in the same individual. PMID- 11391531 TI - Induction of Mdr1b expression by tumor necrosis factor-alpha in rat liver cells is independent of p53 but requires NF-kappaB signaling. AB - The multidrug resistance protein Mdr1b in rats is up-regulated during liver regeneration after partial hepatectomy or after endotoxin treatment. We hypothesize that up-regulation of Mdr1b in these models is TNF-alpha-dependent. The mechanism of Mdr1b activation by TNF-alpha is unknown as TNF-alpha can signal through various pathways, including NF-kappaB and p53, transcription factors for which binding sites in the Mdr1b promoter have been identified. We aimed to elucidate the mechanism of up-regulation of Mdr1b by TNF-alpha. We selectively used constructs expressing dominant negative Fas-associated death domain protein (FADD), TNF receptor associated factor-2 (TRAF2) or IkappaB to inhibit pathways downstream of the TNF receptor. Further, the proteasome inhibitor MG-132 was used, which prevents the breakdown of IkappaB. We show a critical role for NF kappaB in activation of Mdr1b gene expression both in primary rat hepatocytes and in rat hepatoma H-4-II-E cells. Because p53 is up-regulated by TNF-alpha in an NF kappaB-dependent manner and the Mdr1b promoter contains a p53 binding site, we used liver cells expressing a dominant negative p53 to show that TNF-alpha up regulation of Mdr1b is independent of functional p53. Using transient transfection assays, we show that Mdr1b up-regulation correlates with activation of the promoter. Mutation of the NF-kappaB site in the Mdr1b promoter prevents its induction by TNF-alpha. In conclusion our results show that activation of the rat Mdr1b gene by TNF-alpha is a result of NF-kappaB signaling and independent of p53. PMID- 11391532 TI - Caspase inhibition reduces apoptotic death of cryopreserved porcine hepatocytes. AB - Cryopreserved porcine hepatocytes are a ready source of metabolic function for use in a bioartificial liver (BAL). However, cryopreservation is associated with a loss of hepatocyte viability. The mechanism of cell death during cryopreservation is incompletely understood, but may involve apoptosis through caspase activation. This study evaluates the cytoprotective effect of a global caspase inhibitor, benzyloxycarbonyl-Val-Ala-DL-Asp-fluoromethylketone (ZVAD-fmk) during cryopreservation of porcine hepatocytes. Freshly isolated porcine hepatocytes (viability, 97.4% +/- 0.9%) were cryopreserved in 60 micromol/L ZVAD fmk (+ZVAD group) or without ZVAD-fmk (-ZVAD group) for 24 to 72 hours. Apoptotic and necrotic death were both observed after thawing and after 24 hours of culture. Caspase 3-like activity was significantly reduced by ZVAD-fmk, and was associated with improved viability and reduced apoptotic death of porcine hepatocytes after cryopreservation. Mitochondrial membrane potential (MMP) was increased in cultures of porcine hepatocytes that were cryopreserved in ZVAD-fmk. These results demonstrate the following: 1) Caspase 3-like protease activation and apoptosis occurs in porcine hepatocytes during cryopreservation; and 2) mitochondrial injury in this process is reduced by caspase inhibition. PMID- 11391533 TI - Etoposide prevents apoptosis in mouse liver with D galactosamine/lipopolysaccharide-induced fulminant hepatic failure resulting in reduction of lethality. AB - D-Galactosamine (GalN)/lipopolysaccharide (LPS)-induced liver injury is an experimental model of fulminant hepatic failure in which tumor necrosis factor alpha (TNF-alpha) plays a pivotal role. We examined the effects of etoposide on GalN/LPS-induced fulminant hepatic failure. Mice were given an intraperitoneal dose of GalN (800 microg/g body weight)/LPS (100 ng/g body weight) with and without intraperitoneal etoposide (10 microg/g body weight) treatment. Liver injury was assessed biochemically and histologically. TNF-alpha levels in the serum, and apoptosis of hepatocytes and CPP32/caspase-3 in the liver, were determined. GalN/LPS treatment caused lethal liver injury in 87% of animals (13 of 15). The effect was associated with significant increases in TNF-alpha and alanine transaminase (ALT) levels in serum, the number of apoptotic hepatocytes, CPP32/caspase-3 activity, and TNF receptor 1 (TNFR1) mRNA expression in the liver. Etoposide (10 microg/g body weight) was given 3 times (at 50, 26, and 4 hours before GalN/LPS administration). Treatment of GalN/LPS-treated mice with etoposide reduced apoptosis of hepatocytes, resulting in reduction of lethality (13% [2 of 15]), while another topoisomerase II inhibitor, IRCF-193, showed no significant effect. The antilethal effect of etoposide was also confirmed in GalN/TNF-alpha-induced fulminant hepatic failure. Etoposide treatment reduced CPP32/caspase-3 activity in the liver, although it did not alter the serum TNF alpha levels or hepatic TNFR1 mRNA expressions. In addition, etoposide treatment enhanced the mRNA and protein expression of Bcl-xL, an antiapoptotic molecule in the liver. The present findings suggest that etoposide prevents endotoxin-induced lethal liver injury by up-regulation of Bcl-xL, and that etoposide could be useful for the treatment of TNF-alpha-mediated liver diseases. PMID- 11391534 TI - Biliary cholesterol hypersecretion in gallstone-susceptible mice is associated with hepatic up-regulation of the high-density lipoprotein receptor SRBI. AB - Enhanced hepatocellular trafficking of cholesterol to the bile canaliculus and cholesterol hypersecretion appears critical for gallstone formation. Therefore, we studied in more detail the hepatic cholesterol transport pathways in a mouse model of cholesterol gallstone disease. Biliary lipid secretion rates, plasma lipoprotein levels, hepatic expression of lipoprotein receptors, lipid regulatory enzymes, and putative cholesterol transporting proteins were analyzed in gallstone-susceptible C57L/J and gallstone-resistant AKR/J mice, which were fed a lithogenic diet. Biliary cholesterol hypersecretion in C57L mice was associated with decreased plasma high-density lipoprotein (HDL) cholesterol levels and significant hepatic induction of the HDL receptor (SRBI) and cholesteryl ester hydrolase. In response to the lithogenic diet, fatty-acid binding protein of liver (FABPL) was markedly induced in both mouse strains. Caveolin 1 was elevated only in plasma membranes of gallstone-susceptible C57L mice, which also failed to down-regulate cholesterol synthesis. These data suggest a role of the reverse cholesterol transport pathway for genetically determined gallstone susceptibility in the mouse. PMID- 11391535 TI - Alterations in tight junctions differ between primary biliary cirrhosis and primary sclerosing cholangitis. AB - Tight junctions (TJ) of biliary epithelial cells (BEC) and hepatocytes prevent bile regurgitation from the biliary tract. Alterations in these TJs may participate in chronic cholestatic liver diseases such as primary biliary cirrhosis (PBC) and primary sclerosing cholangitis (PSC). We examined the localization of 2 TJ proteins, ZO-1 and 7H6, in these diseases. Frozen sections from livers of PBC, PSC, extrahepatic cholestasis (Ex-C), and hepatitis C associated cirrhosis (LC-C), as well as histologically normal livers, were processed for double-fluorescence immunohistochemistry. In controls and cirrhosis, 7H6 and ZO-1 colocalized surrounding the luminal space of the bile ducts and outlined the bile canalicular spaces between hepatocytes. In untreated PBC, immunostaining for ZO-1 in BEC of bile ducts 40 to 80 microm in diameter was preserved, but that for 7H6 was diminished to absent. In PBC treated with ursodeoxycholic acid (UDCA), immunostaining for 7H6 was well preserved. In PSC as well as in Ex-C, immunostaining for both 7H6 and ZO-1 was well preserved in bile ducts. In hepatocytes, ZO-1 showed preserved immunoreactivity, but immunostaining for 7H6 frequently disappeared. The percentage of bile ducts with immunostaining for 7H6 in all bile ducts with immunostaining for ZO-1 was significantly reduced in PBC compared with that in control, LC-C, Ex-C, and PSC (all P <.0001). Substantial alteration in the TJ protein occurs predominantly in bile ducts in PBC and in hepatocytes in PSC, suggesting increased paracellular permeability along different paracellular routes for bile regurgitation in these chronic cholestatic liver diseases. PMID- 11391536 TI - Differential effects of microsomal enzyme-inducing chemicals on the hepatic expression of rat organic anion transporters, OATP1 and OATP2. AB - The organic anion transporting polypeptides, Oatp1 (Slc21a1) and Oatp2 (Slc21a5), mediate hepatic uptake of cardiac glycosides. Previously, we demonstrated that chemicals that increase cytochrome P450s differentially affect hepatic uptake of cardiac glycosides. We postulated that increased uptake of cardiac glycosides observed after pretreatment of animals with phenobarbital (PB) and pregnenolone 16alpha-carbonitrile (PCN) occurs via increased hepatic expression of Oatp1 and/or Oatp2. Male Sprague-Dawley rats were injected with PB, PCN, 3 methylcholanthrene (3-MC), or vehicle for 4 days. Branched-DNA (bDNA) signal amplification and Western blot analyses were used to assess hepatic Oatp1 and Oatp2 mRNA and protein, respectively. The expression of Oatp1 was not increased by any chemical treatment. Increases in Oatp2 expression were observed from livers of rats treated with PB and PCN, in which PCN caused a robust elevation of Oatp2 mRNA and protein. Oatp2 expression was suppressed in response to 3-MC. To determine the temporal effects of PCN treatment on the expression of Oatp2, rats were administered PCN, livers were extracted at various times, and Oatp2 expression was analyzed. Maximal expression of Oatp2 mRNA was observed at 24 hours and remained elevated, whereas the amount of Oatp2 protein increased throughout the 96-hour interval. The finding that Oatp2 expression increases in response to PB and PCN is consistent with our previous findings that PB and PCN enhance hepatic uptake of cardiac glycosides. These results suggest that Oatp2, but not Oatp1, is inducible by PB and PCN, which imparts the increased capacity of the liver to extract cardiac glycosides from the plasma. PMID- 11391537 TI - Protective immune response to hepatitis C virus in chimpanzees rechallenged following clearance of primary infection. AB - Hepatitis C virus (HCV) infections were evaluated in chimpanzees that had previously cleared HCV and were rechallenged. Animals that had previously cleared HCV infection rapidly cleared homologous and heterologous virus upon rechallenge, indicative of a strong protective immunity. In one animal, sterilizing immunity was observed with regard to viremia, although viral RNA was transiently detected in the liver. Accelerated viral clearance following rechallenge with HCV was observed in animals that had not been exposed to HCV for over 16 years, suggesting that long-lasting protective immunity may be possible. The ability of peripheral blood mononuclear cells (PBMC) to recognize HCV proteins was evaluated during the course of the rechallenge experiments. A very early and strong in vitro recall response to HCV nonstructural proteins appeared to be associated with viral clearance. In contrast, proliferative responses to HCV proteins were not observed in 4 persistently infected chimpanzees, and a weak proliferative response was observed in 1 of 2 animals during acute resolving infection. The results suggest that a strong T-cell proliferative response is induced upon rechallenge of chimpanzees with HCV and that this response is associated with rapid viral clearance. The antibody response to HCV proteins increased by over 1,000-fold in all animals following rechallenge as well. A more complete understanding of the role of the cellular immune response in the clearance of HCV and the nature of the protective immune response following viral clearance may aid in the generation of therapies and vaccines. PMID- 11391538 TI - Inhibition of hepatitis B virus DNA replication by imino sugars without the inhibition of the DNA polymerase: therapeutic implications. AB - Previously we have shown that the imino sugar inhibitor of N-linked glycan processing, N-nonyl-deoxynojirimycin (N-nonyl-DNJ), had antiviral activity in the woodchuck model of chronic hepatitis B virus (HBV) infection. In studying the mechanism of action of this compound, it was discovered that imino sugars could inhibit HBV secretion without inhibiting N-linked glycoprocessing. Although N nonyl-DNJ is an inhibitor of the endoplasmic reticulum (ER) glucosidase, here it is shown that N-nonyl-DNJ retained antiviral activity at concentrations that had no significant impact on ER glucosidase function. Taken together, these results suggested that N-nonyl-DNJ possessed an antiviral activity attributable to a function other than an impact on glycoprocessing. This hypothesis was confirmed by experiments showing that N-nonyl-deoxygalactojirimycin (N-nonyl-DGJ), an alkyl derivative of galactose with no impact on glycoprocessing, retains anti-HBV activity. The data suggest that N-nonyl-DGJ exerts its antiviral action at a point before viral envelopment and may prevent the proper encapsidation of the HBV pregenomic RNA. PMID- 11391539 TI - Intrahepatic hepatitis C viral RNA status of serum polymerase chain reaction negative individuals with histological changes on liver biopsy. AB - For individuals testing anti-HCV positive but negative for HCV RNA in serum, diagnosis remains unclear. Debate exists over whether these individuals have resolved infection or have similar clinical, histological, and virological profiles as serum PCR-positive individuals. The aim of this study was to assess the significance of histological changes in the liver of 33 serum PCR-negative women by investigation of clinical, histological, and intrahepatic HCV RNA status. For comparison, clinical and histological data from 100 serum PCR positive women is presented. Viral RNA status was determined in snap-frozen liver biopsies using a sensitive nested PCR with an internal control. Although serum PCR-positive and -negative individuals shared similar age at diagnosis, source, and duration of infection, they differed from a clinical, histological, and virological perspective. Mean serum ALT levels were significantly lower in serum PCR-negative women (27.4 IU/L +/- 18 vs. 58.7 IU/L +/- 40 P <.001). Similarly, although inflammation (82%) and mild fibrosis (15%) were observed in PCR-negative biopsies, the mean HAI/fibrosis scores were significantly lower than in serum PCR positive biopsies (1.9 +/- 1.5/0.15 +/- 0.4 vs. 4.2 +/- 1.4/1.1 +/- 1.3, respectively). Finally, HCV RNA was not detectable in serum PCR-negative liver biopsies but was detectable in all serum PCR-positive control biopsies. In conclusion, serum PCR-negative individuals may have mild histological abnormalities more suggestive of nonspecific reactive changes, steatosis or nonalcoholic steatohepatitis rather than chronic HCV, even when significant antibody responses are present in serum. Negative serum PCR status appears to reflect cleared past-exposure in liver. PMID- 11391540 TI - Expression of hepatitis C virus NS5A natural mutants in a hepatocytic cell line inhibits the antiviral effect of interferon in a PKR-independent manner. AB - The impact of hepatitis C virus NS5A protein mutations on interferon alfa (IFN alpha) signaling pathway, cell proliferation, and viability is an important issue that is still under debate. We have therefore combined transient and stable expression in a human hepatocytic cell line (Huh7) of 3 full-length NS5A sequences, isolated from patients with or without response to IFN-alpha therapy. Expression of all 3 NS5A-reduced IFN-alpha global antiviral activity on both vesicular stomatitis virus (VSV) and encephalomyocarditis virus (EMCV) replication. We did not show, however, an effect of these 3 NS5A proteins on double-stranded RNA-dependent kinase (PKR) expression and activity as well as colocalization and coimmunoprecipitation between NS5A and PKR. We also failed to show an effect of the 3 NS5A mutants tested on cell proliferation and viability. Overall, our results support an important role of NS5A in controlling IFN-alpha antiviral activity; they show, however, that PKR-independent mechanisms are implicated, at least in liver-derived cells. PMID- 11391541 TI - Protracted, but not acute, hepatitis A virus infection is strongly associated with HLA-DRB*1301, a marker for pediatric autoimmune hepatitis. AB - HLA alleles are known to be associated with susceptibility to develop autoimmune hepatitis (AH), and hepatitis A virus (HAV) infection is postulated as a putative trigger for AH. We investigated whether HLA may influence the outcome of the HAV infection by studying 67 children with self-limited and 39 children with protracted forms of this infection. HLA typing of the uncomplicated forms showed no significant increase of any HLA class I or II alleles. In contrast, DRB1*1301 was present in 46.1% of the children with protracted forms (vs. 9.8% in healthy controls; relative risk [RR]: 7.6; chi(2) = 33.3; P = 2 x 10(-9)). In uncomplicated hepatitis, 45% developed anti-smooth muscle antibody (SMA)/actin antibodies, but only 1 child had detectable antibodies after 3 months of infection onset. In contrast, after 1 year, 69% of the patients suffering protracted forms had titers of anti-SMA/actin antibodies that ranged between 1:40 and 1:160. Within their follow-up, 2 patients developed a Hashimoto's thyroiditis, but the remaining patients showed no signs of developing autoimmune hepatitis. We conclude that the DRB1*1301 haplotype is strongly associated with the protracted forms of HAV infection and suggest that the infection allows a sustained release of liver self-antigens. However, other still-unknown susceptibility genes are required for the full development of pediatric AH. PMID- 11391542 TI - Interferon alfa down-regulates CD81 in patients with chronic hepatitis C. AB - CD81 protein has been shown to bind hepatitis C virus (HCV) envelope 2 (E2) glycoprotein in vitro and may act as a (co)receptor for HCV. Regulation of CD81 expression by interferon alfa (IFN-alpha) and ribavirin could thereby affect the response to antiviral therapy. In the present study, the effects of IFN-alpha and ribavirin on CD81 protein and CD81 mRNA were assessed in peripheral blood lymphocytes (PBL) and isolated human hepatocytes by fluorescence-activated cell sorter (FACS) analysis and real-time polymerase chain reaction (PCR), respectively. In addition, regulation of CD81 in PBL was investigated in 10 patients treated with combination therapy. Incubation with IFN-alpha (50 U/mL) down-regulated total CD81 in PBL (81.7 +/- 11.6% of control; P =.003) and in isolated human hepatocytes (91.6 +/- 8.1% of control; P =.034). Incubation with IFN-alpha with and without ribavirin (2.2 microg/mL) significantly reduced cell surface-associated CD81 protein (83.9 +/- 10.3% of control; P =.003). PBL of untreated patients chronically infected with HCV had significantly higher levels of total CD81 protein compared with PBL obtained from healthy donors (631.1 +/- 93.3 vs. 538.9 +/- 95.2 relative fluorescence units [RFU]; P =.030). Pretreatment cell surface-associated CD81 protein was lower in patients infected with genotype HCV-3 than those infected with HCV-1 (111.8 +/- 15.0 vs. 162.0 +/- 41.3 RFU; P =.019). Furthermore, cell surface-associated CD81 protein was lower 4 weeks after initiation of therapy in patients with an initial virologic response compared with initial virologic nonresponders (110.5 +/- 8.5 vs. 139.8 +/- 27.5 RFU; P =.057). In conclusion, IFN-alpha and ribavirin regulate CD81 expression in vitro and in vivo. CD81 expression correlates with HCV genotype and initial virologic response in patients with chronic hepatitis C. PMID- 11391543 TI - Extended lamivudine treatment in patients with chronic hepatitis B enhances hepatitis B e antigen seroconversion rates: results after 3 years of therapy. AB - A study in Chinese patients with chronic hepatitis B showed that treatment with lamivudine for 1 year significantly improves liver histology and enhances hepatitis B e antigen (HBeAg) seroconversion compared with placebo. Fifty-eight patients from this 1-year study have received long-term treatment with lamivudine 100 mg; the outcome of 3 years of lamivudine is reported here. Before treatment, all patients had detectable HBeAg. HBeAg seroconversion (HBeAg-negative, anti-HBe positive), hepatitis B virus (HBV)-DNA suppression, alanine transaminase (ALT) normalization, emergence of YMDD variant HBV, liver histology, and long-term safety were assessed. After 3 years of continuous treatment with lamivudine 100 mg daily, 40% (23 of 58) of patients achieved HBeAg seroconversion. In patients with baseline serum ALT >2 x upper limit of normal (ULN), the rate of HBeAg seroconversion was 65% (17 of 26). Median serum HBV-DNA concentrations were below the level of detection, and median ALT concentrations were within the normal range throughout 3 years of treatment. YMDD variant HBV emerged in 33 of 58 (57%) patients during the 3 years, of whom 9 (27%) achieved HBeAg seroconversion (6 after emergence of YMDD variant HBV). ALT levels and histologic scores after emergence of YMDD variant HBV did not show major deterioration. Lamivudine was well tolerated during 3 years of therapy. In conclusion, these data in Chinese patients with chronic hepatitis B show enhanced seroconversion rates with extended lamivudine treatment. Up to two thirds of patients with moderately elevated pretreatment ALT achieved HBeAg seroconversion after 3 years of therapy. PMID- 11391545 TI - Adding to the hepatitis B virus treatment arsenal: alpha-glucosidase inhibitor derivatives. PMID- 11391544 TI - Identification of immunodominant hepatitis C virus (HCV)-specific cytotoxic T cell epitopes by stimulation with endogenously synthesized HCV antigens. AB - Hepatitis C virus (HCV)-specific CD8(+) cytotoxic T lymphocytes (CTL) are believed to play an important role in the pathogenesis of liver cell injury and viral clearance in HCV infection. Because HCV does not efficiently infect human cells in vitro and primary infected hepatocytes cannot be used as stimulator/target cells for CTL analysis, development of efficient systems to activate and expand CTL in vitro, reproducing antigen presentation to CTL occurring during natural infection, is mandatory to study CTL activity and to define the hierarchy of immunodominance of CTL epitopes. To achieve this goal, 5 different defective adenoviruses carrying structural and nonstructural HCV genes (core, core-E1-E2, E2, NS3-NS4A, NS3-NS5A) were used to induce the endogenous synthesis of HCV proteins in human adherent mononuclear cells in vitro and to allow their entry into the HLA class I cytosolic pathway of antigen processing. The cytolytic activity of peripheral blood lympho-mononuclear cells (PBMC) from HLA-A2(+) HCV-infected patients stimulated with recombinant adenovirus-infected cells was tested against target cells either pulsed with a panel of synthetic peptides containing the HLA-A2 binding motif or infected with recombinant vaccinia viruses carrying HCV genes. Our study defines a reproducible system to stimulate and expand HCV-specific CTL in vitro that mimics the conditions of antigen encounter in vivo. By this approach, we have identified several HLA-A2 restricted epitopes that should correspond to immunodominant HCV sequences recognized by CTL during natural infection. Therefore, these amino acid sequences represent ideal candidates for the design of therapeutic vaccines for chronic HCV infection. PMID- 11391546 TI - Hepatitis C virus and interferon resistance: it's more than just PKR. PMID- 11391547 TI - Beefing up replication of the hepatitis C virus. PMID- 11391548 TI - Mice and men: are they bile-ologically different? PMID- 11391549 TI - Prevalence of type 2 diabetes mellitus among persons with hepatitis C virus infection in the United States. PMID- 11391550 TI - Mechanism of hepatocyte death after ischemia: apoptosis versus necrosis. PMID- 11391552 TI - The ethics of health care for profit. PMID- 11391554 TI - Going the distance: Mapping and managing polarities. PMID- 11391555 TI - Social justice in a market model world. PMID- 11391556 TI - The University of Iowa Nursing Collaboratory: a partnership for creative education and practice. AB - This article describes a comprehensive partnership between a college of nursing and its corresponding department of nursing services. The Nursing Collaboratory was created to provide an infrastructure for the generation, dissemination, and application of knowledge to improve nursing practice and patient outcomes. In addition to creative problem solving, the Collaboratory serves as an "incubator" of ideas and innovation, engaging nursing staff, faculty, and students in the development of new products and services to enhance nursing education and practice. J Prof Nurs 17:114-120, 2001. PMID- 11391557 TI - Predicting success for baccalaureate graduates on the NCLEX-RN. AB - The purpose of this retrospective study was to identify predictors of success for baccalaureate nursing graduates on the National Council Licensure Examination Registered Nurse (NCLEX-RN). Subjects (505) were graduates of a baccalaureate nursing program in the southeastern United States from 1993 to 1998. The outcome variable was pass/fail on the NCLEX-RN on the first attempt. Predictor variables included type of student (freshman admission, transfer, second degree), age at the time of licensing examination, gender, performance on selected prenursing courses, performance in all junior and senior nursing courses, cumulative grade point average (GPA) at graduation and the Mosby AssessTest score. Results showed a significant relationship between number of Cs, Ds, and Fs in nursing courses and NCLEX-RN results. Students who passed the NCLEX-RN had significantly higher average GPAs, made fewer grades of C or below, and scored higher on the Mosby AssessTest than students who failed. Nontraditional college-age students tended to have a higher passing rate than did traditional age students. A logistic regression model was developed that correctly predicted 76 per cent of the students who failed based on the information available by the end of the end of the first semester of the senior year. J Prof Nurs 17:121-127, 2001. PMID- 11391558 TI - Interaction in graduate nursing Web-based instruction. AB - This article describes interaction and communication methods and issues in the context of two Web-based graduate nursing courses, a theory course and a research course. A conceptual model for graduate nursing Web-based instruction is presented in which learner-centered pedagogic philosophies and interaction techniques are combined to produce active learning at each phase of the Web-based courses. The focus is on asynchronous communication techniques because these represent the dominant communication form in the two courses. Asynchronous communication is interaction in which teacher and student activities are not occurring at the same time and/or place (Berge, 1999). Outcomes of the courses are examined briefly. J Prof Nurs 17:128-134, 2001. PMID- 11391559 TI - Technology-enhanced distance education: from experimentation to concerted action. AB - Despite the rush to embrace technology-enhanced learning (TEL), descriptions of how schools of nursing move toward systemwide implementation of this type of teaching are scarce. There is a wide gap between sporadic dabbling by individual instructors in specific courses and adoption and implementation of TEL throughout a program. This article describes the experiences of a group of nursing faculty who helped move one school of nursing from experimentation with TEL in distance education to concerted action toward a strategic schoolwide plan. J Prof Nurs 17:135-140, 2001. PMID- 11391560 TI - Facilitating scholarship among clinical faculty. AB - This article describes an evolving model of clinical scholarship for clinical track faculty. Contemporary literature regarding scholarship emphasizes broader definitions of scholarship among university faculty, usually with an implicit focus on university faculty with doctoral degrees. Discussions of clinical scholarship focus on scholarship projects with clear application to improved patient care. Clinical-track faculty in university settings serve as exemplars of professional nurse clinicians for their students and for community-based colleagues, and also participate in university life as full faculty. Furthermore, scholarship for clinical faculty is consistent with their participation as academic scholars and as clinical scholars. An important strategy for fostering scholarship among clinical faculty in one school was the creation of a position, Director of Clinical Scholarship, with responsibilities for strengthening organizational support for scholarship activities among clinical-track faculty. Examples of activities and resources designed to foster scholarship are presented, along with preliminary evaluation of scholarship activities of clinical-track faculty. J Prof Nurs 17:141-146, 2001. PMID- 11391561 TI - Nurses' use of tuition waivers: one university's experiences. AB - Although the nursing literature documents the benefits of collaboration between academic and practice settings, the benefits of rewarding clinical preceptors and their organizations has not been examined. In Fall 1996, a large urban public university in New England implemented a program awarding tuition waivers to individual preceptors and clinical agencies who had made significant contributions to the education of nursing students. Three years after implementation, the program was evaluated to determine the extent nurse preceptors and clinical agencies used tuition waivers to assist nurses to enroll in a degree-granting program. Slightly more than one third (36.6 per cent) of the 82 vouchers awarded for a course waiver of tuition and fees were used by 24 nurses. Of the 24 nurses, 12 (50 per cent) were matriculating in the University degree-granting program when they used the voucher. Only one nurse, however, who took her first course on the campus with a voucher, subsequently enrolled in the graduate nursing program during the study period. J Prof Nurs 17:147-150, 2001. PMID- 11391562 TI - The (17)O-NMR shielding range and shielding time scale and detection of discrete hydrogen-bonded conformational states in peptides. AB - The (17)O-NMR shielding range and shielding time scale due to hydrogen-bonding interactions in peptides are critically evaluated relative to those of (1)H-NMR. Furthermore, the assumptions and conclusions in previous (17)O-NMR studies on the detection of discrete conformational states in peptides (V. Tsikaris et al., Biopolymers, 2000, Vol. 53, pp. 135-139) are reconsidered. Consistent examination of the method demonstrates that although (17)O shieldings of peptide oxygens are very sensitive to hydrogen bonding interactions, the (17)O-NMR shielding time scale is not advantageous compared to that of (1)H-NMR, and thus it is not suitable for the detection of discrete hydrogen-bonded conformational states in peptides. (17)O-NMR spectroscopy is prone to interpretation errors due to the formation of (17)O-labeled impurities during the synthetic procedures (A. Steinschneider et al., International Journal of Peptide and Protein Research, 1981, Vol. 18, pp. 324-333). PMID- 11391563 TI - Influence of thermal history on the structural and mechanical properties of agarose gels. AB - Using a multitechnique approach, two temperature domains have been identified in agarose gelation. Below 35 degrees C, fast gelation results in strong, homogeneous and weakly turbid networks. The correlation length, evaluated from the wavelength dependence of the turbidity, is close to values of pore size reported in the literature. Above 35 degrees C, gelation is much slower and is associated with the formation of large-scale heterogeneities that can be monitored by a marked change in the wavelength dependence of turbidity and visualised by transmission electron microscopy. Curing agarose gels at temperatures above 35 degrees C, and then cooling them to 20 degrees C, produces much weaker gels than those formed directly at 20 degrees C. Dramatic reductions in the elastic modulus and failure strain and stress are found in this case as a result of demixing during cure. An interpretation, based on the kinetic competition between osmotic forces (in favor of phase separation) and elastic forces (that prevent it) is proposed. PMID- 11391564 TI - Role of salt bridges in homeodomains investigated by structural analyses and molecular dynamics simulations. AB - Homeodomains are a class of helix-turn-helix DNA-binding protein motifs that play an important role in the control of cellular development in eukaryotes. They fold in a three alpha-helix structural module, where the third helix is the recognition helix that fits into the major groove of DNA. Structural analysis of the members of the homeodomain family led to the identification of interactions likely to stabilize the protein domains. Linking the helices pairwise, three salt bridges were found to be well preserved within the family. Also well conserved were two cation-pi interactions between aromatic and positively charged side chains. To analyze the structural role of the salt bridges, molecular dynamics simulations (MD) were carried out on the wild-type homeodomain from the Drosophila paired protein (1fjl) and on three mutants, which lack one or two salt bridges and mimic natural mutations in other homeodomains. Analysis of the trajectories revealed only small structural rearrangements of the three helices in all MD simulations, thereby suggesting that the salt bridges have no essential stabilizing role at room temperature, but rather might be important for improving thermostability. The latter hypothesis is supported by a good correlation between the melting midpoint temperatures of several homeodomains and the number of salt bridges and cation-pi interactions that connect secondary structures. PMID- 11391565 TI - Effects of structural modifications on some physical characteristics of exopolysaccharides from Lactococcus lactis. AB - The relationship between the primary structure and the chain stiffness of exopolysaccharides (EPSs) and modified EPSs produced by two strains of Lactococcus lactis subsp. cremoris was investigated. The molar mass and radius of gyration of these exopolysaccharides were analyzed by multiangle static light scattering after size-exclusion chromatography. From these results and the chemical structure of the repeating units of the investigated EPSs, the Kuhn lengths could be calculated. We found that the initial Kuhn lengths of the two native EPSs are similar. Modification of the EPSs by removing parts of the side groups resulted in a decrease in both the absolute value and the normalized value of the Kuhn length. It is therefore concluded that partial removal of the side groups of these polysaccharides could make them less efficient as thickeners if no specific interaction with other components occurs. PMID- 11391566 TI - Structural basis for the activity of pp60(c-src) protein tyrosine kinase inhibitors. AB - Conformational searches on three closely related pp60(c-src) protein tyrosine kinase inhibitors of varying potencies were performed to determine a structural basis for their activity. The first was a linear peptide (PDNEYAFFQf), the second its 10-membered cyclic analogue, and the third a cyclic analogue with a para carboxyphenylalanine in place of one the F residues. A common backbone conformation with an antiparallel beta-sheet-like geometry capped by similar beta turns was found for all three peptides, which may be a binding conformation and gives a candidate pharmacophore for further testing. The interaction between some polar side chains and between some of the aromatic rings may be important for maintaining the correct conformation. The differences in potencies of these inhibitors may be attributed to certain thermodynamic and chemical reasons. PMID- 11391567 TI - Influence of solvent and configuration of residues at positions 2 and 3 on distance and mobility of pharmacophore groups at positions 1 and 4 in cyclic enkephalin analogues. AB - The analgesic activity of opioid peptides is mainly connected with their affinity and selectivity for the mu-receptors. The biological activity of cyclic opioid analogues depends on mutual orientation and conformational freedom of aromatic pharmacophore groups at positions 1 and 4. The distance and distance distributions between chromophores at positions 1 [Phe(p-NO(2)), p nitrophenylalanine] and 4 [Nal, beta-(2-naphthyl)alanine], which constitute an energy donor-acceptor pair, were calculated based on measured fluorescence intensity decays of a donor (Nal). The influence of the solvent and configuration of the residues at position 2 and 3 on donor-acceptor distance distribution and mobility of pharmacophore groups at position 1 and 4 in cyclic enkephalin analogues are discussed. PMID- 11391568 TI - Distinguishing malignant from normal oral tissues using FTIR fiber-optic techniques. AB - Oral tissue samples were studied using mid-IR fiber-optic attenuated total reflectance spectroscopy and other spectral techniques. The 1745 cm(-1) band, which is assigned to the ester group (C==O) vibration of triglycerides, is a reliable marker that is present in normal tissues but absent or a weak band in malignant oral tissues. Other bands such as C--H stretching bands and the amide bands are also helpful in distinguishing malignant tissues from normal tissues. Subtraction spectra confirmed the above conclusion. In addition, Raman spectroscopic measurements were in agreement with the results observed from FTIR spectra. PMID- 11391569 TI - Ab initio vibrational calculations on ara-T molecule: application to analysis of IR and Raman spectra. AB - The FTIR and FT-Raman spectra are reported for the arabinonucleoside ara-T (1 beta-D-arabinofuranosylthymine), which shows antiviral activity. The accurate knowledge of the vibrational modes is a prerequisite for the elucidation of drug nucleotide and drug-enzyme interactions. The FTIR and FT-Raman spectra of ara-T were recorded from 4000 to 30 cm(-1). A tetradeuterated derivative (deuteration at N3, and hydroxyl groups O'2, O'3, and O'5) was synthesized and the observed isotopic shifts in its spectra were used for the vibrational analysis of ara-T. The theoretical frequencies and the potential energy distribution (PED) of the vibrational modes of ara-T were calculated using the ab initio Hartree-Fock/3-21G method. An assignment of the vibrational spectra of ara-T is proposed considering the scaled PED and the observed band shifts under deuteration. The scaled ab initio frequencies were in reasonable agreement with the experimental data. PMID- 11391570 TI - FTIR microspectroscopy study of composition fluctuations in extruded amylopectin gelatin blends. AB - The spatial variation in the composition of nonexpanded biopolymer blends prepared by extrusion of mixtures of gelatin with either native or pregelatinized waxy maize starch was studied using a 30-microm aperture FTIR microspectroscopy technique. The ratio of the areas of the "saccharide" bands (953-1180 cm(-1)) and the amide I and II bands (1483-1750 cm(-1)) was used to monitor the relative distributions of the two components of the blend. Two calibration methods were used to obtain amylopectin concentration values from the ratios of the IR bands. The results suggested a high degree of heterogeneity in these blends, despite the thorough mixing expected by twin-screw extrusion processing. The concentration fluctuations were greater for the blends produced by extruding gelatin and native waxy maize starch mixtures. This was in agreement with the reduced degree of conversion of the starch granules when extruded in the presence of gelatin. The FTIR 2-dimensional maps obtained suggested that in the blends produced from either native or pregelatinized starch at all concentrations studied (25/75, 50/50, and 75/25 amylopectin/gelatin) the gelatin constituted the continuous phase. The effect of the spatial resolution on the FTIR microspectroscopy results was considered and the proposed interpretation was verified by the use of polarized light microscopy and FTIR microspectroscopy acquired at higher spatial resolution (10 microm). PMID- 11391571 TI - Interaction of cyanine dyes with nucleic acids. XXI. Arguments for half intercalation model of interaction. AB - The spectral luminescent properties of two groups of monomethine cyanine dyes were studied in the presence of DNA. The first group included five dyes with 5,6 methylenedioxy-[d]-benzo-1,3-thiazole heterocycle and their unsubstituted analogs. Five monomethine pyrylium cyanines and their N-methyl-pyridine analogs were included in the second group. In each pair the pyrylium and pyridine dyes had similar geometry but differed in charge density distribution. The results presented some evidence in favor of the half-intercalation interaction mode between the studied dyes and DNA. When the benzothiazole residue had the lowest electron donor ability between the two heterocycles in the dye molecule, its substitution with the bulky methylenedioxy group led to a significant decrease in fluorescence enhancement of the dye-DNA complex. On the contrary, when the substituents that create steric hindrance (e.g., methylenedioxy and methyl groups) were introduced into the heterocycle with the higher electron donor ability, the fluorescence enhancement value of the dye-DNA complex was virtually unchanged. The changes in the Stock's shift values upon the formation of the dye DNA complexes were in agreement with the proposed half-intercalation model. Interestingly, in the dye-DNA complexes the pyrylium dyes probably resided in a place similar to the pyridine ones. It is possible that the benzothiazole (or benzooxazole) ring intercalated between the DNA bases and the pyrylium (or pyridine) residue was located in the DNA groove closer to the phosphate backbone. PMID- 11391572 TI - Conformational study on poly [gamma-(alpha-phenethyl)-L-glutamate] using vibrational circular dichroism spectroscopy. AB - Vibrational circular dichroism (VCD) and IR absorption spectra are obtained in a chloroform solution for poly[gamma-((R)-alpha-phenethyl)-L-glutamate] (PRPLG) and poly[gamma-((S)-alpha-phenethyl)-L-glutamate] (PSPLG), whose only structural difference is an opposite chiral center in the side chain. Their characteristic amide A, I, and II bands show VCD patterns quite similar to those of poly[gamma benzyl-L-glutamate] (PBLG), indicating that the secondary structure of these polypeptides is a right-handed alpha-helix. The VCD spectra in the CH stretching region exhibit different patterns for PRPLG and PSPLG, reflecting the chirality difference in the side chains. This difference is interpreted on the basis of the additivity of optical activity contributions from the main chain conformation and the chirality difference in the side chains. The results indicate that a VCD difference spectrum of the CH stretching region is a useful diagnostic tool for elucidating local chirality differences. PMID- 11391573 TI - The correlation of Epstein-Barr virus expression and lymphocyte subsets with the clinical presentation of nodular sclerosing Hodgkin disease. AB - BACKGROUND: The pathogenesis of nodular sclerosing Hodgkin disease (HD) has been correlated with Epstein-Barr virus (EBV). The phenotype of lymphocytes in HD and its relations to clinical presentation and to EBV expression have not been characterized fully. Grade II HD is a more aggressive form of the disease. The authors studied cases of HD by flow cytometry (FCM) in an attempt to analyze the phenotype of lymphocytes in the involved lymph nodes and to characterize the phenotype of these lymphocytes in relation to EBV expression, tumor grade, and clinical presentation. MATERIALS AND METHODS The authors prospectively studied lymph nodes from 48 patients with the diagnosis of HD by FCM for T (CD3, CD4, and CD8) and B (CD19) lymphocytes. Ratios of helper to suppressor (CD4 to CD8) and ratios of T to B (CD3 to CD19) lymphocytes were calculated. In situ hybridization for EBV also was performed. The tumors were graded. Clinical data related to age and stage of the disease were retrospectively analyzed. RESULTS: There were 30 male and 18 female patients with an age range of 7 to 77 years (median, 17 yrs). EBV expression was seen in 24 (50%) cases. Eleven (23%) cases were classified as Grade II disease. All Grade II cases showed EBV expression, whereas only 13 (39%) cases of Grade I disease were positive (P = 0.03). EBV-positive cases had a median CD4 to CD8 ratio of 1.62, whereas EBV-negative cases had a ratio of 3.86 (P = 0.01). Grade I cases had a median CD4 to CD8 ratio of 4.58, whereas Grade II cases had a ratio of 1.62 (P = 0.007). EBV-positive cases had a median T lymphocyte to B-lymphocyte ratio of 2.72, whereas EBV-negative cases had a ratio of 3.17 (P = 0.77). Grade I cases had a median T-lymphocyte to B-lymphocyte ratio of 3.51, whereas Grade II cases had a ratio of 1.71 (P = 0.001). A higher percentage of children was seen in the EBV-positive cases than in the negative ones (58% vs. 29%). Cases with low (< 1.5) CD4 to CD8 ratios showed more incidence of high-stage disease (Stages III and IV) than patients with higher ratios (81% vs. 51%). High-stage disease also was seen more frequently in patients with low (< 3) T- to B-lymphocyte ratios (71% vs. 50%). CONCLUSION: The authors found that the local immune response in HD may vary from one case to another. The findings also suggest that EBV may play a role in the pathogenesis of the disease in relation to T- and B-lymphocyte response. A more profound immune suppression and decrease in overall T and helper lymphocytes may be seen in aggressive EBV-positive variants of the disease. These changes may impact the initial presentation of the disease and perhaps its overall biologic behavior. PMID- 11391574 TI - Expression of apoptosis proteins in chronic myelogenous leukemia: associations and significance. AB - BACKGROUND: The mechanisms favoring the growth advantage of Philadelphia chromosome positive cells over normal cells in chronic myelogenous leukemia (CML) are not fully elucidated but could be due partly to altered apoptosis and longer survival of CML clones. Also, little is known about the biologic characteristics of disease progression in CML. Bcl-2 expression has been demonstrated to exert an antiapoptotic effect resulting in increased cell survival. Other proteins such as Bax and Bad are proapoptotic proteins. Fas, a cell surface protein, can be triggered by an appropriate death-promoting ligand (FasL) to activate downstream caspases pivotal in initiation of programmed cell death. Although the mechanisms underlying cellular proliferative and apoptotic pathways are complex, with involvement of multiple interlocking proteins, the relative expression of pro- and antiapoptotic proteins may have an influence on disease progression. This study aimed to determine whether the changes in the cellular expression of Bcl-2, Bax, and Fas correlate with caspase-3 activity and disease progression in CML, or with response to interferon (IFN)-alpha therapy and prognosis in early chronic phase CML. METHODS: Bcl-2, Bax, and Fas expression were measured on whole cell lysates from bone marrow mononuclear cell fractions by Western blot analysis and quantitative radioimmunoassay. Caspase-3 activity was determined using the DEVD system. Specimens from 203 patients with CML were examined. These included 130 patients in early chronic phase disease (ECP; diagnosis to therapy, < or =12 months), 33 patients in late chronic phase (diagnosis to therapy, > 12 months), 27 patients in accelerated phase, and 13 patients in blastic phase. Correlations between apoptosis proteins and CML phases, risk groups in ECP, and response to IFN-alpha therapy and survival in ECP were investigated by standard statistical methods, and positive findings were assessed by multivariate analysis. RESULTS: Levels of Bcl-2, Fas, Bax, and caspase-3 activity did not correlate with disease progression. Among patients in ECP, higher Fas levels correlated with poorer risk groups (P = 0.05) and higher caspase-3 activity correlated with better risk groups (P = 0.048). With IFN-alpha therapy, major cytogenetic responses were noted in 30% of patients with high Fas and 53% with low Fas (P = 0.016) and failure to achieve a complete hematologic response (CHR) in 25% versus 2% (P = 0.0001). Survival was shorter with high Fas levels (5-year rates, 71% vs. 52%; P = 0.002), and the independent poor prognostic significance of high Fas levels was confirmed by multivariate analysis (P = 0.014). Response to IFN-alpha therapy and survival were not significantly different by different levels of Bcl-2, Bax, or caspase-3 activity. CONCLUSIONS: High Fas levels were associated with intrinsically worse disease at diagnosis, whereas high caspase-3 activity was associated with good risk disease. In ECP CML, high Fas levels were associated with significantly worse response to IFN-alpha therapy and with significantly worse survival. The influence of these cellular proteins and caspase-3 activity on apoptosis in CML is complex and merits further investigation. PMID- 11391575 TI - The importance of MUC1 cellular localization in patients with breast carcinoma: an immunohistologic study of 71 patients and review of the literature. AB - BACKGROUND: The MUC1 mucin is present on the apical surface of normal secretory epithelia. In breast carcinoma, MUC1 expression is variable in amount and cellular localization, the significance of which is controversial. The authors undertook a detailed analysis of staining pattern combined with a comprehensive literature review to better understand the role of MUC1 in breast carcinoma. METHODS: Seventy-one patients with breast carcinoma were examined for MUC1, beta catenin, and E-cadherin staining patterns. These data were compared with data from 25 articles from the literature examining the expression of MUC1 in breast carcinoma. RESULTS: All invasive carcinomas showed some MUC1 staining. In invasive ductal carcinomas, MUC1 was detected in the apical membrane (15%), cytoplasm (93%), or circumferential membrane (13%), with 81% of tumors showing a mixture of patterns. Tumors with low overall MUC1 expression (< or = 50% positive tumor cells) had a higher nuclear grade than tumors with high overall MUC1 expression (> 50%; P = 0.01). Tumors with high and low cytoplasmic expression had no difference in nuclear grade (P > 0.3). Circumferential membrane staining was correlated with positive lymph node status (P = 0.011). CONCLUSIONS: In the literature, similar findings prevailed in which overall MUC1 expression was increased in lower grade (10 of 14 studies), estrogen receptor positive (8 of 13 studies) tumors and was associated with a better prognosis (8 of 13 studies). High cytoplasmic staining was associated with a worse prognosis, an association that was not explained by differences in histologic grade. Thus, the presence of MUC1 in the majority of tumor cells is associated with better differentiated tumors and with an improved prognosis. However, aberrantly localized MUC1 in the tumor cell cytoplasm or nonapical membrane is associated with a worse prognosis. PMID- 11391576 TI - Feasibility and low toxicity of early radiotherapy after high-dose chemotherapy and autologous stem cell transplantation for patients with high-risk stage II-III and locally advanced breast carcinoma. AB - BACKGROUND: This prospective trial examined the feasibility, toxicity, and effectiveness of early locoregional radiotherapy after high-dose chemotherapy and autologous stem cell transplantation in patients with high-risk American Joint Committee on Cancer (AJCC) Stage II-III and locally advanced breast carcinoma. METHODS: One hundred forty-seven consecutive patients with high-risk and locally advanced breast carcinoma were included in the current study. All patients received induction chemotherapy with a doxorubicin-based therapy, which was consolidated with high-dose cyclophosphamide, carboplatin, and thiotepa followed by autologous stem cell support. Within 50 days of the transplant, the patients were treated with locoregional radiotherapy that included the chest wall or breast, the axilla and supraclavicular area, and the internal mammary chain. The volume of lung included in the treatment volume was kept to a minimum. The central lung distance of the tangential fields ranged from 0.6-2.0 cm (mean, 1.1 cm). Tamoxifen was given based on receptor status. RESULTS: One hundred forty-six of 147 patients received the planned treatment. Only six patients had a delay in the initiation of radiotherapy, and another 16 patients had delays during radiotherapy. Leukocyte and platelet toxicities during radiotherapy were not life threatening and blood counts thereafter returned to normal. Grade 2 (according to National Cancer Institute Common Toxicity Criteria) skin toxicity occurred in 22% of patients and Grade 3 skin toxicity occurred in 6% of patients. Radiation pneumonitis was reported to occur in 5 patients (< 4%). After a median follow-up of 36 months from diagnosis (range, 6-64 months), there were no long-term organ toxicity and no secondary malignancy reported. No treatment-related deaths were reported. Three patients (< 3%) developed locoregional recurrence. CONCLUSIONS: Locoregional radiotherapy after high-dose chemotherapy and autologous stem cell transplantation appears to be feasible and can be delivered safely within 10 weeks of transplantation. The short-term and long-term toxicity are reported to be low, with good local control. PMID- 11391577 TI - Clinical characterization of pulmonary large cell neuroendocrine carcinoma and large cell carcinoma with neuroendocrine morphology. AB - BACKGROUND: Large cell carcinoma has been classified as four potential types based on its neuroendocrine morphology and evidence of neuroendocrine differentiation discernible by immunohistochemistry or electron microscopy. However, the clinical relation among these four categories has not been clearly defined. In 1999, the World Health Organization (WHO) categorized large cell neuroendocrine carcinoma as a variant of large cell carcinoma. MATERIAL AND METHODS The authors analyzed 119 cases of large cell carcinoma from a total of 2070 primary lung carcinoma cases resected surgically between 1969-1999. Using light microscopy, electron microscopy, and immunohistochemical staining, the authors reclassified these cases into large cell neuroendocrine carcinoma (LCNEC), large cell carcinoma with neuroendocrine differentiation (LCCND), large cell carcinoma with neuroendocrine morphology (LCCNM), and classic large cell carcinoma (CLCC). RESULTS: In multivariate analyses, the authors found that large cell carcinoma with neuroendocrine features, which combined LCNEC, LCCND, and LCCNM, impacted both the overall survival and disease-free survival of patients. The clinical behavior of LCCNM was similar to that of LCNEC. CONCLUSIONS: Large cell carcinomas with neuroendocrine features appear to be more clinically aggressive than CLCCs. The authors' findings suggest that the histologic identification of neuroendocrine features in tumor tissue from patients diagnosed with large cell carcinoma of the lung may have clinical relevance. PMID- 11391578 TI - The spleen as a diagnostic specimen: a review of 10 years' experience at two tertiary care institutions. AB - BACKGROUND: Few studies have examined the yield of the diagnostic splenectomy, and the relevance of these studies to the management of patients with unexplained splenomegaly or a splenic mass are limited by low number of cases, the use of selection criteria, and the lack of modern terminology and modern ancillary studies. The current study correlates clinical intent with preoperative clinical and radiologic studies and histologic findings in an assessment of the diagnostic yield of splenectomy. METHODS: The medical charts, laboratory data, radiologic studies, and pertinent preoperative biopsies on all patients who underwent splenectomy between the years 1986 and 1995 were reviewed, and the clinical intent behind the procedure was correlated with histologic findings. RESULTS: One hundred twenty-two of the 1280 patients underwent splenectomy for diagnosis, and in 116 patients a specific disease was identified histologically that explained the splenomegaly/splenic mass; malignancy was the most common cause of unexplained splenomegaly or splenic mass, though benign neoplasms and reactive disorders were documented in 25% of the cases. Primary splenic lymphomas were most commonly of large cell B-cell type. CONCLUSIONS: In the setting of splenomegaly or splenic mass, splenectomy has a high diagnostic yield and usually discloses a malignancy. The clinical category of "primary splenic lymphoma" is biologically heterogeneous, and the diagnosis is usually an intermediate grade (not low grade) lymphoma. The range of conditions associated with splenic masses were quite commonly associated with diseases that are amenable to fine-needle aspiration (FNA) diagnosis, whereas those disorders associated only with splenomegaly included a large fraction of diseases for which FNA may yield either incomplete or misleading results. PMID- 11391579 TI - Combined etoposide, ifosfamide, and cisplatin in the treatment of patients with advanced thymoma and thymic carcinoma: an intergroup trial. AB - BACKGROUND: Patients with thymic tumors (thymoma and thymic carcinoma) are known to respond to a variety of chemotherapeutic agents, including single-agent ifosfamide and cisplatin with etoposide. The purpose of this trial was to evaluate the response rate, progression free survival, overall survival, and toxicity of combined etoposide, ifosfamide, and cisplatin (VIP) in patients with advanced thymoma and thymic carcinoma. METHODS: From July 1995 through February 1997, 34 patients with advanced thymoma or thymic carcinoma were entered on trial to receive etoposide (75 mg/m2 on Days 1-4) ifosfamide (1.2 g/m2 on Days 1-4), and cisplatin (20 mg/m2 on Days 1-4). Cycles were repeated every 3 weeks for four total cycles. RESULTS: Among 28 evaluable patients (pathology review excluded 6 patients), there were no complete responses and 9 partial responses (complete and partial responses combined, 32%; 95% confidence interval, 16-52%). The median follow-up was 43 months (range, 12.8-52.3 months), the median duration of response was 11.9 months (range, < 1-26 months), and the median overall survival was 31.6 months. Based on Kaplan-Meier estimates, the 1-year and 2-year survival rates were 89% and 70%, respectively. The toxicity was predominantly myelosuppression. CONCLUSIONS: The VIP regimen has moderate activity in patients with advanced thymic malignancies. However, with limited follow-up, the results of this trial appear to be inferior to other chemotherapy regimens reported in large Phase II trials performed in patients with this disease. PMID- 11391580 TI - Adjuvant 5-fluorouracil plus doxorubicin in D2-3 resected gastric carcinoma: 15 year experience at a single institute. AB - BACKGROUND: The authors evaluated the efficacy of adjuvant chemotherapy with 5 fluorouracil (5-FU) plus doxorubicin in gastric carcinoma after D2-3 curative resection. They also evaluated the effect of dose-related factors (delivered total dose/m(2), actual dose intensity [ADI], relative dose intensity [RDI]) of this regimen on patient survival. METHODS: A total of 301 patients with Stage II to IV (en bloc resected T4b; 1984 American Joint Committee on Cancer staging) were accrued between 1984 and 1996. Chemotherapy was started within 4 weeks of surgery according to the following schedule: intravenous bolus injection of doxorubicin 40 mg/m2 every 3 weeks for 12 cycles and 5-FU 400 mg/m2 weekly for 60 weeks. The toxicity and survival were evaluated. RESULTS: The median follow-up duration was 58 months. Sixty-four percent of the total patients and 71.7% of the patients who did not experience recurrence during the chemotherapy finished the protocol completely with acceptable toxicities. The 5- and 10-year disease free survival rates of total 301 patients were 58.4% and 46.5%, and the overall survival rates were 62.1% and 50.5%, respectively. Treatment completion group showed survival benefit over the early termination group in 5-year survival (75.2% vs. 52.9%; P = 0.0005). The median ADI of 5-FU and doxorubicin were 349 and 11 mg/m2/week, and the median RDIs of 5-FU and doxorubicin were 0.87 and 0.83, respectively. Multivariate analysis demonstrated that completion of chemotherapy is an independent prognostic factor of both disease free and overall survival. However, ADI and RDI did now show any effect on survival. CONCLUSIONS: Adjuvant chemotherapy with 5-FU plus doxorubicin for 60 weeks after D2-3 dissection induced promising survival duration with acceptable toxicities. Full administration of the planned dosage of the combined drugs is recommendable as opposed to early termination of the chemotherapy in gastric carcinoma. PMID- 11391581 TI - Expression of survivin correlates with apoptosis, proliferation, and angiogenesis during human colorectal tumorigenesis. AB - BACKGROUND: To identify the role of survivin, a novel inhibitor of apoptosis (IAP) in colorectal tumorigenesis, the authors investigated tissue expression of survivin in human colorectal tumors including 43 hyperplastic polyps, 171 adenomas with low dysplasia, 42 adenomas with high dysplasia, and 60 carcinomas in adenoma, and examined whether the expression of survivin correlated with tumor cell apoptosis, proliferation, and angiogenesis, which is known to initiate the imbalance between cell proliferation and apoptosis. METHODS: Immunohistochemical staining for the paraffin sections by using the monoclonal antibodies, survivin, p53, bcl-2, Ki-67, and CD34, was performed by the standard avidin-biotin peroxidase technique. The apoptotic cells were detected by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling method, using an Apop Tag in situ detection kit. RESULTS: The immunoreactivity of survivin significantly increased in the transition from adenoma with low dysplasia to high dysplasia/carcinoma (P < 0.001). Similar changes in protein expression were observed for p53 but not for bcl-2, which was expressed throughout the colorectal tumorigenesis. This transition was associated with a significant decrease in the apoptotic index (AI) and significant increases in the Ki-67 labeling index (LI) and microvessel density (MVD; P < 0.001 for both). The expression of survivin inversely correlated with AI and was positively correlated with Ki-67 LI and MVD (P < 0.001 for both). CONCLUSIONS: These results suggest that, like p53, survivin plays an important role in transition from adenoma with low dysplasia to high dysplasia during human colorectal tumorigenesis. PMID- 11391582 TI - Correlation between progression free survival and response rate in patients with metastatic colorectal carcinoma. AB - BACKGROUND: All endpoint measures currently used to evaluate chemotherapeutic treatment benefit in clinical trials of patients with metastatic colorectal carcinoma (MCRC) have disadvantages. Overall survival (OS) is the most objective parameter but is flawed as an efficacy criterion, partly because potentially active second-line treatments are not controlled in many Phase III studies, and measured OS may be influenced (positively or negatively) to an unknown degree by second-line therapy. Measuring progression free survival (PFS) may be a means of isolating the real impact of first-line regimens. METHODS: To test this hypothesis, the authors analyzed pooled data from all Phase III studies on first line treatment in patients with MCRC reported from 1990 through 2000 for correlations between PFS, response rate (RR), and OS in MCRC patients that may suggest PFS as an improved indicator of the efficacy of first-line treatment. Spearman rho correlation coefficients (r) and regression equations were used. RESULTS: The authors analyzed data from 29 studies involving nearly 13,500 patients. Significant correlations were found between PFS and RR (r = 0.655; P < 0.0001; a 10% RR increment corresponded to a 1-month increase in PFS), between RR and OS (r = 0.408; P = 0.0009; an 11.4% RR increment corresponded to a 1-month increase in OS), and between PFS and OS (r = 0.481; P < 0.0001; a 1-month increase in PFS corresponded to a 0.68-month increase in OS). CONCLUSIONS: PFS is the most appropriate primary endpoint for MCRC studies, because it can express the antitumor activity of a first-line chemotherapy regimen regardless of measures used after disease progression. PFS deserves further evaluation as an endpoint measure. PMID- 11391583 TI - A Phase I/II study of weekly paclitaxel and 3 days of high dose oral estramustine in patients with hormone-refractory prostate carcinoma. AB - BACKGROUND: The maximum tolerated dose (MTD) and efficacy of weekly 1-hour paclitaxel with 3 days of high dose oral estramustine were evaluated in patients with hormone-refractory prostate carcinoma. METHODS: Patients enrolled in cohorts of three received two cycles of six weekly treatments with 1 week of rest: Cohort I received paclitaxel 40 mg/m2 and estramustine 600 mg/m2, and Cohorts II-IV received paclitaxel 60 mg/m2, 75 mg/m2, or 90 mg/m2, respectively, and estramustine 900 mg/m2. Toxicity was assessed weekly, and response was measured by serum prostate specific antigen (PSA), abdominal computed tomography scans, and bone scans at Week 13. RESULTS: Eighteen patients were enrolled, with 12 in Cohorts III and IV. Four patients did not complete treatment. Grade 3 toxicity included one patient with nausea and diarrhea in Cohort III and one patient each with neutropenia and edema followed by Grade 4 thromboembolism in Cohort IV. Grade 1-2 anemia or myelotoxicity were not observed; 3 patients had neuropathy, 5 patients had hair loss, and 8 patients had gastrointestinal symptoms. A decline in the serum PSA level > or = 50% occurred in none of three patients, one of three patients, four of six patients, and four of six patients in Cohorts I-IV, respectively. An intent-to-treat analysis showed responses in 9 of 18 patients (50%) in Cohorts I-IV, with 9 of 15 responders (60%) in Cohorts II-IV. Seven patients achieved declines in serum PSA levels > 75%. The median duration of PSA response was 16.7 weeks. Response was observed in one of three patients with measurable disease. CONCLUSIONS: The MTD for 1-hour weekly paclitaxel was 90 mg/m2 with 3 days of 900 mg/m2 estramustine. Hematologic and neurotoxicity were reduced markedly, and gastrointestinal symptoms were ameliorated, but thromboembolic events were unaffected. PSA response rates were within the expected 60% range for these agents. PMID- 11391584 TI - Prospective assessment of voiding and sexual function after treatment for localized prostate carcinoma: comparison of radical prostatectomy to hormonobrachytherapy with and without external beam radiotherapy. AB - BACKGROUND: Voiding and sexual function after treatment are major determinants of quality of life in prostate carcinoma patients. Erectile dysfunction, incontinence, and urinary symptoms, both obstructive and irritative, have a significant negative impact on patient quality of life. This prospective study was undertaken to evaluate voiding, sexual function, and their impact on patients with localized prostate carcinoma who were treated with radical retropubic prostatectomy (RP) and to compare these patients with patients who were undergoing hormonobrachytherapy with external bean radiotherapy (HBTC) and patients who were undergoing hormonobrachytherapy without external beam radiotherapy (HBT). METHODS: Patients treated for localized prostate carcinoma with either RP or interstitial palladium-103 (103Pd) HBTC or HBT were prospectively administered a voiding and sexual function questionnaire before any treatment was initiated and at posttreatment visits. Questionnaire components included the American Urological Association Symptom Score (AUASS) and specific items that addressed urinary control and sexual function from the University of California at Los Angeles Prostate Cancer Index. Questionnaire results were compiled, and differences among treatment groups were assessed over time. RESULTS: From January 1997 to November 1999, 127 consecutive patients were treated with either unilateral or bilateral nerve-sparing RP (42 patients), HBTC (40 patients) or HBT (45 patients) by 2 surgeons proficient in all procedures. Using the overall score and the obstructive subscale (OAUA) of the AUASS, the RP group showed a posttreatment decrease in scores compared with both HBTC and HBT groups. OAUA scores of HBTC and HBT groups were significantly greater than scores in RP patients over the course of the study. HBTC patients had increased irritative symptoms initially when compared with RP patients, and, although not statistically significant, the magnitude of the difference persisted over the course of the study. Total AUASS and subscale scores for the RP group returned to near baseline levels within 12 months. The use of incontinence pads was a criterion for urinary incontinence, and the proportion of patients returning to baseline continence was lower in RP patients over the course of the study. No notable differences in Voiding Bother (VB) scores were found. Initially RP patients experienced worse Sexual Function (SF) scores; however, scores for RP patients changed over time and approached the levels seen in HBTC patients at 18 months. The Sexual Function Bother (SFB) scores also were higher initially in the RP group but then decreased to similar levels observed for HBTC patients by 18 months. None of the treatment groups returned to near baseline SF or SFB scores during the course of this study. CONCLUSIONS: Comparison of voiding function indicated that HBTC and HBT patients initially have more obstructive voiding symptoms, whereas urinary incon- tinence is initially worse in RP patients. Initially RP patients demonstrated worse SF and SFB scores, but RP patients returned to HBTC levels within 18 months. PMID- 11391585 TI - Clinicopathologic and familial characteristics of endometrial carcinoma with multiple primary carcinomas in relation to the loss of protein expression of MSH2 and MLH1. AB - BACKGROUND: The frequency of synchronous or metachronous multiple primary carcinomas in patients with endometrial carcinoma has been reported to be between 10% and 23% and is highest among all gynecologic carcinomas. However, clinical characteristics and underlying genetic abnormalities in endometrial carcinoma with multiple primary carcinomas has not been well clarified. Endometrial carcinoma is the most commonly associated extracolonic malignancy in hereditary nonpolyposis colorectal carcinoma in which germ line mutations in DNA mismatch repair genes, particularly in MSH2 and MLH1, are known to cause this syndrome. The purpose of the current study was to investigate clinicopathologic and familial characteristics including MSH2, MLH1, and p53 expression in endometrial carcinoma with multiple primary carcinomas, by comparing them to endometrial carcinoma without other primary malignancies. METHODS: Patients were divided into two groups: 30 patients with synchronous or metachronous multiple primary carcinomas other than endometrial carcinoma and 116 patients with endometrial carcinoma without other primary malignancies. Clinicopathologic characteristics, family history of cancer, and immunohistochemical protein expression of MSH2, MLH1, and p53 expression were investigated in both groups, and 15 endometria from benign disease were used for normal controls in immunohistochemistry. RESULTS: The frequency of high risk clinicopathologic factors of endometrial carcinoma and 5-year survival rates and the frequency of p53 overexpression were not statistically different between the two groups. However, the loss of MSH2 and/or MLH1 expression was significant in endometrial carcinoma with multiple primary carcinomas, when compared with endometrial carcinoma alone (22 of 30 vs. 31 of 116). In cases with multiple primary carcinomas, particularly those diagnosed before the patient was 55 years of age or those in which the patient had a family history of cancer, the frequency of this loss was especially high (11 of 13 and 10 of 11, respectively). CONCLUSIONS: The clinical or biologic nature of endometrial carcinoma with multiple primary carcinomas seems to be similar to endometrial carcinoma alone. A high incidence of defective MSH2 and MLH1 protein in endometrial carcinoma with multiple primary carcinomas, however, suggests that abnormalities in the function of MSH2 and MLH1 may play an important role in tumorigenesis for patients with endometrial carcinoma with multiple primary carcinomas and their families. PMID- 11391586 TI - Diagnosis, pathology, staging, treatment, and outcome of epithelial ovarian neoplasia in patients age < 21 years. AB - BACKGROUND: Epithelial ovarian neoplasms are rare in patients under the age of 21 years. This is a report of a series of such patients documenting their presentation, histologic type, stage of disease, treatment, and outcome. METHODS: Clinical findings, histology, stage, treatment, and outcomes of 19 patients with epithelial ovarian neoplasia are reported. All histology was rereviewed. RESULTS: The median age at the time of diagnosis was 19.7 years (range, 14.1-21.8 years), and the median follow-up was 5.6 years (range, 0.2-19.5 years). The most common presenting symptom was dysmenorrhea (100%) followed by abdominal pain (68%), and the initial diagnosis usually was made ultrasonographically. There were nine (47%) serous tumors, 7 (37%) mucinous tumors, 2 (11%) small cell carcinomas, and 1 (5%) endometrioid carcinoma. Seventy-nine percent of tumors were unilateral, and 84% were low malignant potential or well differentiated tumors. Surgical treatment included unilateral salpingo-oophorectomy in 12 patients (63%), total abdominal hysterectomy and bilateral salpingo-oophorectomy in 6 patients (32%), and ovarian cystectomy in 1 patient (5%). Fifteen patients (79%) had Stage I disease, and 4 patients (21%) had Stage III disease at the time of diagnosis. There were two deaths in this series, and both occurred in patients with small cell anaplastic carcinoma. CONCLUSIONS: Epithelial ovarian neoplasias are rare in patients in this age group but must be included in the differential diagnosis of an ovarian mass. Most patients present with Stage I tumors of low malignant potential. In these patients, good survival is achieved with unilateral salpingo oophorectomy and preservation of fertility. In contrast, small cell carcinomas are very aggressive, and patients with this variant require intensive therapy. PMID- 11391587 TI - Full dose reirradiation combined with chemotherapy after salvage surgery in head and neck carcinoma. AB - BACKGROUND: The purpose of this study was to analyze the tolerance and efficacy of full dose reirradiation combined with chemotherapy in patients with head and neck carcinoma (HNC) with a high risk of recurrence after salvage surgery. METHODS: Between 1991 and 1996, 25 patients having a recurrence or a second primary tumor in a previously irradiated area (> 45 grays [Gy]) were entered in this prospective study. All of them received salvage surgery and had a positive surgical margin and/or lymph node involvement with capsular rupture (N+R+). Adjuvant radiochemotherapy was given, delivering 60 Gy per 30 fractions with concomitant hydroxyurea and 5-fluorouracil. The median total cumulative dose of the 2 irradiations was 118 Gy. The median follow-up after the second irradiation was 66 months. RESULTS: During the reirradiation course, Grade 3 and 4 mucositis were observed in 40% and 12%, respectively. Analysis of late effects (> 6 months after reirradiation) showed that 16% of the patients had osteoradionecrosis and 40% had Grade 2-3 cervical fibrosis (Radiation Therapy Oncology Group scoring system). The patterns of failure were as follows: local only (n = 9), lymph node only (n = 2), local and lymph node only (n = 1), and metastatic (n = 4). The 4 year survival rate after reirradiation was 43% (95% confidence interval, 25-62). CONCLUSIONS: Full dose reirradiation combined with chemotherapy after salvage surgery in high risk patients with HNC was feasible with an "acceptable" toxicity and led to a relatively good 5-year survival rate. These results prompted the authors to initiate a multicentric randomized trial that is ongoing (GETTEC GORTEC 99-01) to evaluate the benefit of adjuvant radiochemotherapy in these types of patients. PMID- 11391588 TI - The accuracy of head and neck carcinoma sentinel lymph node biopsy in the clinically N0 neck. AB - BACKGROUND: Sentinel lymph node (SLN) biopsy originally was described as a means of identifying lymph node metastases in malignant melanoma and breast carcinoma. The use of SLN biopsy in patients with oral and oropharyngeal squamous cell carcinoma and clinically N0 necks was investigated to determine whether the pathology of the SLN reflected that of the neck. METHODS: Patients undergoing elective neck dissections for head and neck squamous cell carcinoma accessible to injection were enrolled into our study. Sentinel lymph node biopsy was performed after blue dye and radiocolloid injection. Preoperative lymphoscintigraphy and the perioperative use of a gamma probe identified radioactive SLNs; visualization of blue stained lymphatics identified blue SLNs. A neck dissection completed the surgical procedure, and the pathology of the SLN was compared with that of the remaining neck dissection. RESULTS: Sentinel lymph node biopsy was performed on 40 cases with clinically N0 necks. Twenty were pathologically clear of tumor and 20 contained subclinical metastases. SLNs were found in 17 necks with pathologic disease and contained metastases in 16. The sentinel lymph node was the only lymph node containing tumor in 12 of 16. CONCLUSIONS: The SLN, in head and neck carcinomas accessible to injection without anesthesia, is an accurate reflector of the status of the regional lymph nodes, when found in patients with early tumors. Sentinel lymph nodes may be found in clinically unpredictable sites, and SLN biopsy may aid in identifying the clinically N0 patient with early lymph node disease. If SLNs cannot be located in the neck, an elective lymph node dissection should be considered. PMID- 11391589 TI - Angiography effectively supports the diagnosis of hepatic metastases in medullary thyroid carcinoma. AB - BACKGROUND: Medullary thyroid carcinoma (MTC) belongs in the group of neuroendocrine tumors with early lymphatic and hepatic dissemination. A high rate of undetectable metastases is hypothesized to be responsible for the frequent mismatch between the apparent relatively small tumor burden and the elevated plasma tumor marker level. METHODS: Thirty-six MTC patients with residual/recurrent biochemical signs (elevated basal calcitonin level) and/or characteristic general symptoms (diarrhea and/or flushing) were systematically examined by conventional radiology, whole-body 18F-deoxyglucose positron emission tomography (PET), dynamic liver computed tomography and magnetic resonance imaging, and hepatic angiography. RESULTS: Conventional diagnostic imaging revealed lymph node (LN) involvement in the cervical, mediastinal, supraclavicular, and axillary regions (16 cases), and multiple pulmonary (3 cases), bony (1 solitary and 1 multiple case), and breast (1 case) metastases. (18)F-deoxyglucose PET identified all these extralymphatic metastatic lesions (except 2 cases with multiple pulmonary metastases), and also supradiaphragmatic LN involvement in 34 (94%) patients. In 32 (89%) cases, multiple small (generally < or = 1 cm) hypervascular, hepatic metastases undetectable by other imaging methods were localized angiographically. Of the 23 original pathologic specimens investigated, 18 (78%) exhibited LN involvement. The smallest primary tumor in patients with hepatic metastases was 1 cm. CONCLUSIONS: Hepatic angiography is recommended for primary staging in MTC patients with a primary tumor measuring 1 cm or larger, and/or pathologically proven LN involvement, and also during restaging for suspected recurrences to avoid unnecessary extensive surgical LN dissection in the neck and mediastinum. PMID- 11391590 TI - P21WAF1/CIP1 messenger RNA expression in hepatitis B, C virus-infected human hepatocellular carcinoma tissues. AB - BACKGROUND: The primary objective of this study was to clarify the significance of p21WAF1/CIP1(p21) gene expression in the tumorgenicity of hepatitis B virus (HBV) and hepatitis C virus (HCV) infected human hepatocelluar carcinoma (HCC). METHODS: The authors performed Northern blot hybridization to compare the p21 messenger (m) RNA expression levels among 16 HCC cases. They detected tissue HBVx mRNA (Northern blot) and plus- and minus-strand HCV RNA (reverse transcription polymerase chain reaction) in liver tissues. They also measured alanine transaminase (ALT) levels and indocyanine green retention rate at 15 minutes (ICG R15). RESULTS: The p21 transcripts of tumor (T) tissues could be identified with lower intensity than nontumor (N) tissues in all 4 HBVx mRNA(+) cases, 8 of 10 HCV RNA(+) cases, and 1 of 3 B(-), C(-) cases (1 case was positive for both viruses). p21 mRNA expression levels of N tissues were significantly higher in HCV RNA(+) cases than in HBVx mRNA(+) cases. p21 mRNA expression levels of N tissues were significantly correlated with serum ALT levels. CONCLUSIONS: In HCV hepatitis, p21 mRNA expression is up-regulated to control cell cycle under regeneration stress. Once the liver develops HCC, the p21 mRNA expression decreases to prominently low levels. The up-regulated p21 expression may play a role as a guard to prevent hepatocytes from tumorgenicity in HCV hepatitis. PMID- 11391591 TI - Familial pulmonary carcinoid tumors. AB - BACKGROUND: Pulmonary carcinoid tumors are rare and usually occur sporadically. Infrequently, they arise in association with multiple endocrine neoplasia type 1 (MEN1). Familial pulmonary carcinoid tumors not associated with MEN1 have not been described. METHODS: Two sets of first-degree relatives diagnosed with primary pulmonary carcinoid tumors with no clinical features of MEN1 were identified in a pair of siblings and in a mother and daughter. Mutations in the MEN1 gene were sought using polymerase chain reaction analysis on paraffin embedded tissue from two members of one of the families. RESULTS: Histopathologic and immunohistochemical studies confirmed the diagnoses of carcinoid tumors. None of these patients and no family members had features of MEN1. DNA analysis did not detect germline mutations in the MEN1 gene. CONCLUSIONS: The occurrence of familial pulmonary carcinoid tumors in the absence of MEN1 suggests a novel, rare germline mutation specific to the development of pulmonary carcinoids. PMID- 11391592 TI - A micromorphometry-based concept for routine classification of sentinel lymph node metastases and its clinical relevance for patients with melanoma. AB - BACKGROUND: The sentinel lymph nodes (SLNs) as the primary targets for lymphatic metastases can be removed selectively by gamma probe-guided sentinel lymph nodectomy (SLNE) in nearly all patients with cutaneous melanoma. Correspondingly high standards in terms of specificity, sensitivity, and microstaging are required for the evaluation of SLNs. METHODS: Since 1995, the authors have performed SLNE in 389 lymph node regions (LNRs) on 342 patients with melanoma. The harvested 636 SLNs and a further 1394 nonsentinel lymph nodes (non-SLNs) were evaluated by standardized, semiserial section histology, including immunohistochemistry. For each LNR, this technique permitted routine S classification using two simple morphometric parameters: the number of tumor involved, 1-mm slices of the SLNs (n) and the centripetal depth of metastatic cell invasion (d). S1 was defined by 1 < or = n < or = 2 and d < or = 1 mm, equivalent to localized peripheral tumor cell deposits; S2 was defined by n > 2 and d < or = 1 mm, indicating more extended peripheral metastases; S3 was defined by d > 1 mm in SNLs with deeper metastatic infiltration; and S0 meant no detectable tumor cells (n = 0). RESULTS: The authors diagnosed 325 SLNs as S0, 24 SLNs as S1, 22 SLNs as S2, and 18 SLNs as S3. The occurrence of at least one melanoma-positive non-SLN subsequent regional completion lymph node dissection (RCLND) significantly increased from 0 of 12 in S1 SLNs to 2 of 13 in S2 SLNs and 9 of 15 in S3 SLNs (P = 0.001; chi-square test). Like the T classification of the primary melanoma, the S classification proved to be a highly significant predictor for distant metastasis (P < 0.001). It turned out to be an independent factor of influence on distant metastasis and survival in multivariate Cox analyses, which included tumor thickness, primary tumor site, patient gender, and patient age as covariates. CONCLUSIONS: The data presented recommend the S staging concept as a promising option to fill a gap between the T and conventional N component of the pTNM classification. If its predictive capacity can be confirmed in multicenter studies, then the S classification may become the decisive criterion for or against RCLND, and a combined T plus S staging system will help to improve prognostically relevant stratification of melanoma patients in adjuvant therapy trials. PMID- 11391593 TI - Cerebrospinal fluid gastrin releasing peptide in the diagnosis of leptomeningeal metastases from small cell carcinoma. AB - BACKGROUND: The clinical diagnosis of leptomeningeal metastases is often difficult to substantiate. Patients with an underlying malignancy typically present with neurologic symptoms referable to multiple levels of the neuraxis. Although most patients have an abnormal cerebrospinal fluid (CSF), less than 60% have evidence of malignant cells on cytologic examination from a single lumbar puncture, and the disease is usually advanced in patients with positive results. An elevated serum level of gastrin releasing peptide (GRP) in patients with small cell carcinoma has emerged as one of the most useful markers for disease activity. METHODS: A patient with small cell carcinoma presented with signs of meningitis and an abnormal CSF. However, the CSF gave repeatedly negative cytologic results. Hence, serum and CSF were analyzed for GRP. RESULTS: The CSF GRP level was elevated by more than six orders of magnitude above the serum level. An autopsy demonstrated extensive meningeal and parenchymal brain involvement by small cell carcinoma. CONCLUSIONS: The diagnosis of leptomeningeal metastases in patients with small cell carcinoma can be established by CSF GRP testing, even when cytologic examination is negative. PMID- 11391594 TI - Clinical sensitivity of p53 mutation detection in matched bladder tumor, bladder wash, and voided urine specimens. AB - BACKGROUND: Mutations in the p53 tumor suppressor gene may correlate with an increased risk of recurrence and disease progression in patients with bladder carcinoma. The ability to accurately and sensitively detect p53 mutations in cytology specimens may be of benefit in the treatment of bladder carcinoma patients with superficial, minimally invasive disease. METHODS: Genomic DNA was isolated from 49 cases, each of which was comprised of matched bladder tumor tissue, bladder wash, and voided urine specimens obtained concurrently at a single institution. The genomic DNA was analyzed for mutations in the p53 tumor suppressor gene using a p53 mutation detection assay. Automated dideoxy sequencing of mutant specimens also was performed. RESULTS: Of the 49 cases, 29 (59%) showed no evidence of p53 mutations in the tumor, bladder wash, or voided urine specimens. Of the remaining 20 cases, 19 showed evidence of mutations in the tumor. Of these 19 p53 mutant bladder tumors, 16 (84%) were detected in the matched bladder wash and 16 (84%) were detected in the matched voided urine specimens. One case resulted in the detection of mutant p53 in the voided urine and the bladder wash, but not in the tumor. Analysis of the results between tumor tissue and bladder wash or tumor and voided urine showed 84.2% sensitivity, 96.8% specificity, and 91.8% accuracy. Sequence analysis of the mutant cases showed that the mutations detected in the tumor tissue were the same mutations detected in the bladder wash and the voided urine specimens. CONCLUSIONS: Both voided urine and bladder wash specimens from patients with bladder carcinoma were found to provide a high rate of clinical accuracy for the determination of the p53 gene status in patients with bladder tumors. PMID- 11391595 TI - The number of intratumoral dendritic cells and zeta-chain expression in T cells as prognostic and survival biomarkers in patients with oral carcinoma. AB - BACKGROUND: Dendritic cells (DCs) are antigen-presenting cells with a unique ability to cross prime T cells and generate strong antitumor responses. This study evaluates the presence and prognostic significance of DCs as well as functional T cells, which accumulate in the microenvironment in patients with oral squamous cell carcinoma (OSCC). METHODS: Immunohistochemistry for S-100 positive or p55 positive DCs and for T-cell receptor (TcR)-associated zeta-chain expression in tumor-infiltrating lymphocytes (TILs) was performed in 132 paraffin embedded specimens from patients with primary OSCC. The median clinical follow-up for the patients was 50 months. The numbers of intratumoral DCs or TILs expressing the zeta chain were determined microscopically and compared with clinical and pathohistologic prognostic parameters, including disease stage, T stage or tumor grade, lymph node involvement, as well as disease free survival and overall survival. RESULTS: Immunostaining identified DCs in the epithelial compartment of the tumors (S-100 positive) as well as interdigitating reticular DCs (p55 positive) in peritumoral areas. Based on S-100 staining, intratumoral DC infiltrates were low (<10 DCs per high-power field [HPF]) in 20% of OSCC specimens, intermediate (10-20 DCs per HPF) in 42% of OSCC specimens, and high (>20 DCs per HPF) in 37% of OSCC specimens. The number of S-100 positive DCs was positively and significantly correlated with that of p55 and of TILs with normal zeta-chain expression. A low number of infiltrating S-100 positive DCs was more predictive of poor survival (hazard ratio, 7.95) than lymph node involvement (hazard ratio, 3.36) or late T stage (hazard ratio, 2.92). A significant but weaker association of p55 positive DC infiltration with survival was observed. Low density of DCs and low or absent expression of the zeta chain in TILs correlated with each other and predicted the poorest survival and the greatest risk. CONCLUSIONS: The number of DCs infiltrating the tumor is a highly significant prognostic parameter in patients with OSCC. Furthermore, the absence or paucity of DCs is strongly linked to abnormalities in the TcR-associated zeta chain in TILs. The two biomarkers, zeta-chain expression in TILs and the number of S-100+ DCs in the tumor, independently predict overall survival, disease free survival, and time to disease recurrence in patients with OSCC. PMID- 11391596 TI - Increased genetic damage in oral leukoplakia from high risk sites: potential impact on staging and clinical management. AB - BACKGROUND: Two staging systems for oral leukoplakias have been proposed to better predict prognosis. Although one system includes site as an independent determinant, its use is controversial. METHODS: Recent studies have shown that loss of heterozygosity (LOH) in oral premalignancies is associated with risk of progression. The authors analyzed 127 oral dysplasias for LOH on 3 chromosome arms (3p, 9p, and 17p). The lesions included 71 from the floor of mouth, ventrolateral tongue, and soft palate complex (designated high risk [HR] sites) and 56 from the rest of the oral cavity (low risk [LR] sites). RESULTS: Dysplasias from HR sites contained significantly higher LOH frequencies than LR sites (percentage with any loss, P = 0.0004; percentage with multiple losses, P = 0.0001; percentage loss on each of the arms, P < 0.05). Loss on 3p and/or 9p, a pattern associated with a 24-fold increased risk of progression (Rosin MP, Cheng X, Poh C, Lam WL, Huang Y, Lovas J, et al. Use of allelic loss to predict malignant risk for low-grade oral epithelial dysplasia. Clin Cancer Res 2000;6:357-62) was more frequent among HR lesions (P = 0.0005). Loss of heterozygosity frequencies were elevated at HR sites among both genders and among smokers and nonsmokers. For different histologic groups, LOH frequencies were elevated for HR sites in mild dysplasias (P < 0.05) and moderate dysplasias (marginal significance, P = 0.06), but not in severe dysplasias/carcinoma in situ. CONCLUSIONS: Anatomic location of mild and moderate oral dysplasias in Western populations may be an important diagnostic indicator because lesions at HR sites have a greater tendency to include genetic alterations associated with elevated risk of progression. PMID- 11391597 TI - Acquisition of secondary structural chromosomal changes in pediatric ewing sarcoma is a probable prognostic factor for tumor response and clinical outcome. AB - BACKGROUND: The Ewing sarcoma (ES) group of tumors commonly have the t(11;22)(q24;q12) or other rearrangements involving 22q12. In addition to these consistent aberrations, both numeric and structural aberrations have been reported: namely gains of chromosomes 8 and 12, the unbalanced translocation t(1;16), and deletions at the short arm of chromosome 1. METHODS: To evaluate the frequency and to study the prognostic implications of some of these aberrations in children, the authors performed a pilot study of 26 ES pediatric patients by classic cytogenetics and/or interphase fluorescence in situ hybridization (FISH) and compared these data with clinical parameters. RESULTS: Gains of chromosomes 8 and 12 were detected, by interphase FISH, in 48% (10 of 21) and 38% (6 of 16) of the tumors, respectively, and this was not significant with respect to treatment response. Statistical analysis revealed that the presence of additional secondary structural chromosomal aberrations was associated with an unfavorable outcome (P = 0.0034 as an independent prognostic value as an unfavorable marker). Presence of metastasis at diagnosis also was found to be associated with poor outcome (P = 0.0131). Spectral karyotyping analysis was shown to facilitate the detection of more complex structural chromosomal aberrations in a representative ES tumor. CONCLUSIONS: It is important to determine whether additional structural chromosomal aberrations are present in ES tumors because it appears that a more complex karyotype with multiple chromosomal aberrations is associated with poor outcome in ES. PMID- 11391598 TI - Only pathologic complete response to neoadjuvant chemotherapy improves significantly the long term survival of patients with resectable esophageal squamous cell carcinoma: final report of a randomized, controlled trial of preoperative chemotherapy versus surgery alone. AB - BACKGROUND: Surgery is the standard treatment for patients with resectable esophageal carcinoma, but the long term prognosis of these patients is unsatisfactory. Some randomized trials of preoperative chemotherapy suggest that the prognosis of patients who respond may be improved. METHODS: This randomized, controlled trial compared patients with clinically resectable esophageal epidermoid carcinoma who underwent surgery alone (Arm A) with those who received preoperative chemotherapy (Arm B). Overall survival and the prognostic impact of major response to chemotherapy were analyzed. Forty-eight patients were enrolled in each arm. Chemotherapy consisted of two or three cycles of cisplatin (100 mg/m2 on Day 1) and 5- fluorouracil (1000 mg/m2 per day continuous infusion on Days 1-5). In both study arms, transthoracic esophagectomy plus two-field lymphadenectomy was performed. The two groups were comparable in terms of patient characteristics. RESULTS: Forty-seven patients were evaluable in each arm. The curative resection rate was 74.4% (35 of 47 patients) in Arm A and 78.7% (37 of 47 patients) in Arm B. Treatment-related mortality was 4.2% in both arms. The response rate to preoperative chemotherapy was 40% (19 of 47 patients), including 6 patients (12.8%) who achieved a pathologic complete responses. Overall survival was not improved significantly. The 19 patients in Arm B who responded to chemotherapy and underwent curative resection had significantly better 3-year and 5-year survival rates (74% and 60%, respectively) compared with both nonresponders (24% and 12%, respectively; P = 0.0002) and patients in Arm A who underwent complete resection (46% and 26%, respectively; P = 0.01): Patients who achieved a pathologic complete response (P = 0.01), but not those who achieved a partial response (P = 0.2), had significantly improved survival. CONCLUSIONS: Patients with resectable esophageal carcinoma who underwent preoperative chemotherapy and obtained a pathologic complete response had a significantly improved long term survival. Major efforts should be undertaken to identify patients before neoadjuvant treatments who are likely to respond. PMID- 11391600 TI - Complementary therapies in palliative cancer care. AB - BACKGROUND: Complementary medicine has become an important aspect of palliative cancer care. This overview is primarily aimed at providing guidance to clinicians regarding some commonly used complementary therapies. METHODS: Several complementary therapies were identified as particularly relevant to palliative cancer care. Exemplary studies and, where available, systematic reviews are discussed. RESULTS: Promising results exist for some treatments, e.g. acupuncture, enzyme therapy, homeopathy, hypnotherapy, and relaxation techniques. Unfortunately, the author finds that the evidence is not compelling for any of these therapies. CONCLUSION: These results point to some potential for complementary medicine in palliative care. They also demonstrate an urgent need for more rigorous research into the value (or otherwise) of such treatments in palliative and supportive cancer care. PMID- 11391599 TI - Chemotherapy in patients with prostate specific antigen-only disease after primary therapy for prostate carcinoma: a phase II trial of oral estramustine and oral etoposide. AB - BACKGROUND: A Phase II study was initiated to evaluate the effectiveness of an oral regimen of etoposide and estramustine in patients with early recurrent prostate carcinoma. METHODS: Patients with early recurrent prostate carcinoma as indicated by an increasing prostate specific antigen (PSA) level and without any evidence of metastatic disease were treated with oral etoposide 50 mg/m2/day and estramustine 15 mg/kg/day in divided doses for 21 days, followed by a 7-day rest period. Patients received a maximum of four cycles. RESULTS: Eighteen patients were entered in this study. The median serum PSA was 3.1 (range, 0.3-30.3) at the time of entry into the trial. Sixteen patients were assessable for response. Serum PSA declined to undetectable levels in 13 patients with 2 additional patients meeting the criteria for partial response; the median duration of response was 8.5 months (range, 1-18 months). Most patients developed gastrointestinal, cardiac, or hematologic complications. Grade 3 toxicities included neutropenia (one patient), deep venous thrombosis (three patients), and chest pain (one patient). One patient developed acute myelogenous leukemia (French-American-British, acute myelogenous leukemia M5) 23 months after initiating the chemotherapy. CONCLUSIONS: The combination of oral etoposide and oral estramustine resulted in a high rate but only a short duration of response in patients with early recurrent prostate carcinoma. The regimen was poorly tolerated, and the toxicity was significant. This regimen should not be considered standard therapy for the treatment of early recurrent prostate carcinoma, but further exploration of treatment in this setting is warranted. PMID- 11391601 TI - Soft tissue sarcoma--compliance with guidelines. AB - BACKGROUND: Because soft tissue sarcomas (STS) are rare, guidelines for the diagnosis and treatment of patients with STS were developed. Because the diagnostic management is essential for definitive treatment, adherence to these guidelines is important. METHODS: Primary STS registered by the Comprehensive Cancer Center North-Netherlands from January 1989 to January 1996 were analyzed retrospectively with regard to adherence to the diagnostic guidelines. Urogenital, gastrointestinal STS, and Kaposi sarcomas were excluded. RESULTS: Three hundred fifty-one STS patients were analyzed. In the specialized center, 69% of patients were age < 60 years, whereas, in district hospitals, 63% of patients were age > 60 years. With increased age, referral to the center declined in a linear fashion. For all guidelines, adherence was significantly better in the center. In district hospitals, patient volume had no significant influence on compliance with the guidelines, except for the management of patients with STS > or = 3 cm. In district hospitals, where fewer than 15 patients were treated in the 7-year period, significantly more often, an inadequate biopsy or even no biopsy procedure was performed prior to resection. CONCLUSIONS: In many aspects of the diagnostic process of STS, existing guidelines were not followed, especially in community hospitals. Adherence to all individual guidelines was significantly better in the specialized center. To improve compliance with future STS guidelines, appropriate guideline development, dissemination, and implementation programs should be developed. Concentration of patients with STS in a limited number of hospitals and intensified collaboration with specialized centers seem advisable. Special attention should be paid to older patients, who significantly more often were not referred to a specialized center. PMID- 11391602 TI - Predicting prostate carcinoma volume and stage at radical prostatectomy by assessing needle biopsy specimens for percent surface area and cores positive for carcinoma, perineural invasion, Gleason score, DNA ploidy and proliferation, and preoperative serum prostate specific antigen: a report of 454 cases. AB - BACKGROUND: DNA ploidy analysis of prostate carcinoma is a generally accepted prognostic marker, particularly when tumors are extraprostatic at the time of surgery. In the past decade, the DNA content of prostate carcinoma frequently has been assessed in needle biopsy specimens based on the assumption that ploidy, in conjunction with serum prostate specific antigen (PSA) and Gleason score, provides valuable pretreatment information. METHODS: Between 1995 and 1998, the authors identified a consecutive series of 454 prostate carcinomas, verified by needle biopsies and followed by radical retropubic prostatectomies (RRP). Based on the needle biopsies, DNA ploidy and MIB-I immunostaining were measured by digital image analysis (DIA). The authors also quantified the percent of nuclei in four categories from the DNA histograms. The DIA data were combined with the age of the patient at diagnosis, the serum PSA, Gleason score, percent cores and percent surface area positive for carcinoma, and status of perineural invasion in multivariate models using tumor volume and risk of extraprostatic extension (EPE) at RRP as the outcome variables. RESULTS: Joint predictors of tumor volume at RRP were the percent cores positive for carcinoma (P < 0.0001), serum PSA (P < 0.0001), the percent surface area positive for carcinoma (P < 0.0001), and the percent nuclei classified by DNA quantification to be in the "S-phase" category (P = 0.03). Joint predictors of risk of EPE were the percent cores positive for carcinoma (P = 0.0004), a Gleason score of 7 (P < 0.0001), a Gleason score of 8 or 9 (P < 0.0001), serum PSA (P = 0.006) and perineural invasion (P = 0.02). CONCLUSIONS: After adjusting for traditional prognostic markers, DNA ploidy interpretation and MIB-I quantitation of prostate carcinoma did not appear to jointly predict either outcome variable in the multivariate models. However, a quantitative measure related to both ploidy and proliferation, the percent of nuclei in the putative "S-phase" category from the DIA histograms, was found to jointly predict for tumor volume. PMID- 11391603 TI - Cervical screening by neural networks. PMID- 11391604 TI - Less medical intervention after sharp demarcation of Grade 1-2 cervical intraepithelial neoplasia smears by neural network screening. AB - BACKGROUND: Neural network technology has been used for the daily screening of cervical smears in The Netherlands since 1992. The authors believe this method might have the potential to demarcate diagnoses of Grade 1-2 cervical intraepithelial neoplasia (CIN 1-2). METHODS: Of 133,196 women who were screened between 1992-1995, there were 2236 CIN 1-2 smears; 1128 of which were detected by means of neural network screening (NNS) (n = 83,404 women) and 1108 of which were diagnosed by conventional screening (n = 49,792 women). Cytologic and clinical outcomes (first cytologic or histologic follow-up diagnosis) were retrieved for all the women in the study population (n = 1920). Stratification based on clinical outcome resulted in the cases being grouped as overdiagnosed, concordant, or underdiagnosed. The smears were performed by general practitioners, whereas the biopsies were obtained by gynecologists. RESULTS: The prevalence rate for CIN 1-2 was 1.15% (95% confidence interval [95% CI], 1.08 1.23%) for NNS and 1.92% (95% CI, 1.80-2.04%) for conventional diagnosis (P < 0.001). Concordance with histology was significantly higher for NNS (53.9%; 95% CI, 50.7-57.0%) compared with conventional screening (29.2%; 95% CI, 26.4-32.2%). In addition, overdiagnosis was significantly lower for cases diagnosed by NNS (39.4%; 95% CI, 36.3-42.4%) compared with cases diagnosed by conventional screening (62.4%; 95% CI, 59.3-65.5%). CONCLUSIONS: Neural network-based screening can lead to fewer women being burdened unnecessarily with a cytologic diagnosis of CIN 1-2 by resulting in a sharp demarcation in these diagnoses and a corresponding reduction in unnecessary medical interventions. [See editorial on pages 171-172, this issue.] PMID- 11391605 TI - Cytologic artifacts and pitfalls of thyroid fine-needle aspiration using ThinPrep: a comparative retrospective review. AB - BACKGROUND: The ThinPrep Processor has gained popularity as a collection and preparation technique for fine-needle aspiration biopsy (FNAB). Specific cytologic criteria to evaluate ThinPrep preparation (TP) may differ from those of conventional preparation (CP). The authors retrospectively reviewed the quality, cytologic features, and pitfalls of TP versus CP in thyroid FNABs and addressed the cytomorphologic criteria used to evaluate TP specimens. METHODS: Thyroid FNABs received between January 1996-July 1999 were identified from the computer files of the Department of Pathology, University of Michigan (Ann Arbor, MI). Histologic correlation and clinical follow-up were reviewed. The cytology slides were reevaluated for cellularity, cellular preservation, artifacts, background material, architectural integrity, cytoplasmic details, and nuclear details by two observers. RESULTS: Of the 209 thyroid FNABs performed during the study period, TP and CP prepared 127 and 82 cases respectively. Histologic correlation was available in 68 (33%) cases (32 TP and 36 CP). Overall sensitivity was 80% and specificity was 98%. The sensitivity of CP versus TP was 87% and 70%, respectively. Thyroid FNABs prepared by TP, as compared with CP, were characterized by the following: The TP slide 1) allowed assessment of the overall specimen cellularity but not individual passes of an FNAB, 2) contained only "hard" colloid that appeared dense, markedly fragmented, or in droplets, 3) showed crowded, tight, tissue clusters with loss of cellular preservation, especially in the larger aggregates, 4) demonstrated more cell shrinkage, 5) showed increased disruption of the cytoplasm and numerous naked nuclei, 6) occasionally gave nucleoli a more prominent appearance, and 7) was less likely to show nuclear grooves and "pseudoinclusions" in papillary carcinoma. CONCLUSIONS: This study concluded that cytologic features used to evaluate thyroid FNABs prepared by CP may need to be modified when using TP. Awareness of the above described findings and further studies to evaluate TP are essential to avoid potential diagnostic pitfalls. PMID- 11391606 TI - Transthoracic fine-needle aspiration biopsy of pulmonary spindle cell and mesenchymal lesions: a study of 61 cases. AB - BACKGROUND: Spindle cell and mesenchymal lesions of the lung encompass a wide variety of benign and malignant conditions. However, to the authors' knowledge, because of their rarity, few reports concerning their cytologic findings are available in the literature. The current review emphasizes the cytomorphologic features, differential diagnosis, and potential pitfalls associated with these lesions. METHODS: Seven hundred seventy-nine percutaneous lung fine-needle aspiration (FNA) specimens were retrieved from the authors' cytopathology files over a period of 5 years. Sixty-one cases (7.8%) in which a spindle cell component was the dominant or key feature were identified. The authors reviewed the cytologic smears, immunocytochemical studies, and corresponding surgical material and clinical information. RESULTS: Of these 61 aspirates, 33 (54%) were reactive processes (31 granulomas, 1 organizing pneumonia, and 1 inflammatory pseudotumor). Five cases (0.8%) were benign neoplasms (2 hamartomas, 2 solitary fibrous tumors, and 1 schwannoma). Twenty-three cases (38%) were malignant neoplasms (8 cases were primary tumors [including 5 carcinomas with spindle cell or sarcomatoid features, 1 spindle cell carcinoid tumor, 1 leiomyosarcoma, and 1 synovial sarcoma] and 15 cases were secondary tumors [including 9 melanomas, 2 leiomyosarcomas, 1 malignant fibrous histiocytoma, 1 meningioma, 1 sarcomatoid renal cell carcinoma, and 1 uterine malignant mixed mullerian tumor]). A specific diagnosis was rendered in 52 cases (85%). No false-positive cases were encountered but there was one false-negative case. One patient who was diagnosed with granulomatous inflammation on FNA was found to have nonsmall cell lung carcinoma on subsequent transbronchial biopsy. No malignant cells were identified in the smears on review. The FNA from the organizing pneumonia was interpreted as a solitary fibrous tumor whereas the inflammatory pseudotumor was diagnosed as granulomatous inflammation. The FNA from one pulmonary hamartoma initially was considered to be nondiagnostic. One solitary fibrous tumor and the schwannoma were diagnosed as smooth muscle tumor and spindle cell tumor, not otherwise specified, respectively. Among the malignant tumors, the primary synovial sarcoma and one of the metastatic malignant melanomas initially were interpreted as primitive neuroectodermal tumor/Ewing sarcoma and poorly differentiated carcinoma, respectively. CONCLUSIONS: Spindle cell lesions of the lung rarely are encountered on transthoracic lung FNA and are comprised of a wide variety of benign and malignant entities. By correlating clinical and radiologic data, cytologic findings, and ancillary studies, a high diagnostic accuracy rate can be achieved with FNA. PMID- 11391607 TI - Ultrasound-guided fine-needle aspiration biopsy of the thyroid bed. AB - BACKGROUND: Ultrasound (US) has been shown to be a sensitive technique for monitoring patients for recurrent thyroid carcinoma in the thyroid bed after total thyroidectomy. However, the role of US-guided fine-needle aspiration biopsy (FNAB) in the confirmation of sonographically indeterminate or suspicious masses has not been adequately addressed. The purposes of this study were to determine the sensitivity and specificity of US-guided FNAB of the thyroid bed for diagnosing recurrent carcinoma after total thyroidectomy and to highlight potential diagnostic pitfalls. METHODS: Twenty-one patients with a history of total thyroidectomy and histologically confirmed thyroid carcinoma who had undergone US-guided FNAB of hypoechoic lesions in the thyroid bed were included in this retrospective study. Fifteen of the 21 had papillary carcinoma (PC), 5 had medullary carcinoma (MC), and 1 had Hurthle cell carcinoma (HTC). The cytologic features of the aspirates were compared with histopathologic findings of pre- and post-FNA surgery. Immunohistochemical staining for thyroglobulin, calcitonin, and parathyroid hormone was performed in four cases. RESULTS: The cytologic diagnosis from the US-guided FNABs was conclusive in 20 of 21 cases. Fifteen cases were diagnosed as recurrent tumor (12 PC, 2 MC, and 1 HTC), and 13 of the 15 were confirmed subsequently by histology. Five cases were diagnosed as benign (two residual benign thyroid tissue, one parathyroid gland [PG] tissue, and two reparative changes) and hence were not resected. There was one false positive diagnosis in which PG was misdiagnosed as PC. Immunohistochemical studies helped to confirm the diagnosis of PG tissue in two cases and of MC in two cases. The sensitivity of US-guided FNA for diagnosing recurrent carcinoma in the thyroid bed after total thyroidectomy was 100% and the specificity was 85.7%. CONCLUSIONS: US-guided FNAB was found to be a sensitive and specific test for diagnosing sonographically indeterminate lesions in the thyroid bed. One potential diagnostic pitfall was the misdiagnosis of normal residual thyroid or PG tissue as recurrent tumor. Careful attention to cytologic details and the use of selected immunohistochemical staining may help to prevent these misdiagnoses. PMID- 11391608 TI - Fine-needle aspiration characteristics of hibernoma. AB - BACKGROUND: Hibernoma is a rare, benign lipomatous tumor with features of brown fat. The preoperative diagnosis of hibernoma is difficult at times because its clinical, radiographic, and fine-needle aspiration (FNA) characteristics overlap with those of liposarcoma. METHODS: The preoperative FNA findings of eight surgically excised hibernomas from seven patients (three men and four women, ages 24-60 years) were reviewed. The cytologic features were compared with the histologic features of the corresponding surgical specimens as well as lipomatous tumors and other lesions that may cause confusion in the differential diagnosis. RESULTS: The FNA cytologic features of the hibernomas were found to correspond well with their histologic appearance. The FNA findings included small, round, brown fat-like cells with uniform, small cytoplasmic vacuoles and regular, small, round nuclei; delicate branching capillaries; and variable numbers of mature fat cells. CONCLUSIONS: The FNA cytologic features of hibernoma are characteristic and useful in the preoperative investigation of lipomatous tumors, particularly with regard to excluding a diagnosis of liposarcoma. PMID- 11391609 TI - Cytologic evaluation of cyclin D1 expression in primary breast carcinoma. AB - BACKGROUND: Preoperative assessment of the biologic characteristics of primary breast carcinoma is important because neoadjuvant medical therapy is being used increasingly. In the current study, the authors attempted to evaluate the validity of cyclin D1 assay in fine-needle aspiration (FNA) samples from patients with primary breast carcinoma. METHODS: FNA samples were obtained prior to therapy and multiple slides were stored at -80 degrees C for subsequent immunocytochemical analysis (ICA). ICA for cyclin D1 protein was performed on FNA samples from 51 breast carcinoma patients and 20 samples from patients with benign breast disease. In 45 breast carcinoma patients who had undergone surgery, sections were taken from paraffin blocks and stained by ICA for cyclin D1 validation. Possible correlations between cyclin D1 expression in the FNA samples and the biologic data of the patients also were analyzed. RESULTS: Cyclin D1 expression was detected in 37 FNA samples from 51 breast carcinomas (72.5%) whereas expression of cyclin D1 was detected in 8 FNA samples from 20 patients with benign breast disease (40%). In histologic sections after surgery, 26 cases of breast carcinoma (65%) showed a positive reaction to cyclin D1. Concordance for the presence of cyclin D1 between FNA samples and histologic samples was 75%. Cyclin D1 expression was high in patients with the tumors that expressed estrogen receptor (ER) (30 of 34 vs. 5 of 11; P = 0.028) and progesterone receptor (PR) (33 of 38 vs. 2 of 7; P = 0.007). There was no significant relation found between cyclin D1 expression and tumor size or lymph node metastasis. Cyclin D1 expression within invasive ductal carcinoma was observed in > 80% of low or intermediate nuclear grade tumors but its expression decreased to 61.5% (8 of 13 cases) in tumors with high nuclear grade (P = 0.023). All 14 breast carcinomas in which the S-phase fraction was 15% showed cyclin D1 expression. Cyclin D1 expression was found to be correlated inversely with proliferative activity in breast carcinoma (P = 0.023). CONCLUSIONS: The results of the current study show that cyclin D1 expression can be measured by ICA in FNA samples with reasonable concordance with the results of histologic section. Cyclin D1 expression was found to be associated with ER/PR status and cell differentiation. The results of the current study indicate that the measurement of novel molecular markers could be performed adequately in FNA samples as well as in histologic sections. PMID- 11391610 TI - Uroplakin as a marker for typing metastatic transitional cell carcinoma on fine needle aspiration specimens. AB - BACKGROUND: Immunohistological markers specific for a single type of epithelium are rare. Recently, urothelium tissue-specific genes were cloned. The genes encoded a family of transmembrane proteins, uroplakins, that are expressed only in urothelial mucosa. Using uroplakin antibodies on paraffin-embedded tissue, a previous study demonstrated positive staining in 66% of metastatic transitional cell carcinoma (TCC) cases and negative staining in all other tumors (including breast, ovarian, lung, and gastrointestinal carcinomas) tested. The current study addresses the diagnostic value of uroplakins in conventional fine-needle aspiration (FNA) material in establishing a diagnosis of metastatic TCC. METHODS: Representative slides from 27 FNA cases of metastatic TCC and 52 non-TCC carcinomas were collected. The avidin-biotin-peroxidase method was utilized, using polyclonal antiuroplakin as the primary antibody on 95% ethanol-fixed, Papanicoloau-stained direct smears. RESULTS: Twenty-five of 27 metastatic TCC cases (93%) were found to stain positively for uroplakin with a superficial membrane/microluminal staining pattern. A few cells with diffuse membranous staining also were noted in 48% of the positive metastatic TCC cases. The superficial membrane/microluminal staining pattern was not observed in any of the non-TCC carcinomas. However, approximately 6% of these cases (3 of 52 cases) did show rare tumor cells with diffuse membranous staining. CONCLUSIONS: The application of uroplakin antibodies to 95% ethanol-fixed FNA direct smears has improved the sensitivity of the antibody for metastatic TCC while maintaining a specificity comparable to that of paraffin-embedded tissue. The authors believe that these antibodies have diagnostic potential in cytopathology in the evaluation of metastatic TCC. PMID- 11391611 TI - DNA cytometry confirms the utility of the Bethesda system for the classification of Papanicolaou smears. AB - BACKGROUND: Developed in 1989, the Bethesda System has largely replaced previous classifications of Papanicolaou (Pap) smears from the uterine cervix. The system is binary, dividing smears into two groups - low-grade, squamous, epithelial lesions (LSIL) or high-grade, squamous, epithelial lesions (HSIL). A third category, atypical squamous cells of undetermined significance (ASCUS), is used to classify minimal cellular changes that do not satisfy the criteria for the low or high-grade categories. This study was designed to confirm the utility of this binary division and to compare the results with another classification system (the Munich II Nomenclature) that is not binary but contains three divisions or grades for dysplasia - low, intermediate, and high. METHODS: Pap smears were obtained from 593 women with a cytologic diagnosis of dysplasia based on the Munich System. Smears were then classified by the Bethesda System into LSIL or HSIL. Patients were followed for 2 years either with biopsy or repeat cytology. The initial smears were restained by the Feulgen method, and ploidy was evaluated by interactive DNA cytometry. RESULTS: Of 241 cases of LSIL, 39% were diploid, 57% polyploid, and 4% aneuploid. Of 352 cases classified HSIL, 4% were diploid, 17% polyploid, and 79% aneuploid. After 2 years of follow-up, 2 of 108 patients who were biopsied and who were originally classified as diploid progressed to cervical intraepithelial neoplasia/carcinoma in situ (CIN/CIS) whereas 109 of 217 patients who were aneuploid and biopsied were found to have CINIII/CIS. CONCLUSIONS: The two divisions of the Bethesda System, LSIL and HSIL, correlated with ploidy as evaluated by cytometry. Aneuploidy was found to be useful to separate cases of HSIL from those of LSIL as defined in the Bethesda System. Because of the binary division, use of a system with three divisions for dysplasia, such as the Munich II Nomenclature, creates a therapeutic dilemma because a single diagnostic category (usually the intermediate grade) may contain both self-limiting and progressive lesions. DNA cytometry of Pap smears was found to be useful as a routine procedure. PMID- 11391612 TI - DNA image analysis combined with routine cytology improves diagnostic sensitivity of common bile duct brushing. AB - BACKGROUND: Cytologic evaluation of common bile duct brushings has a low sensitivity for diagnosing malignancy because of scant cellularity, poor cellular preservation, or sampling errors occur. The aim of this study was to evaluate whether cytology combined with image analysis improves the diagnostic accuracy of bile duct brushing in comparison with cytology alone. METHODS: Forty-nine specimens of bile duct brushings obtained from 45 patients during endoscopic retrograde cholangiopancreatography were evaluated using cytology and image analysis. Specimens were classified as negative, atypical, suspicious, or malignant by using cytologic evaluation. DNA histograms were classified as diploid (D), broad diploid (BD), aneuploid (A), or tetraploid (T). Degree of hyperploidy (DH), representing cells with a DNA content > 5C was evaluated using a cutoff value of > or = 1%. Final diagnosis of cancer was based on tissue specimens that were obtained by fine-needle aspiration or surgical biopsy and clinical fol- low-up. RESULTS: Thirty-four patients ultimately proved to have a malignancy. Cytology revealed 19 negative cases, 15 atypical cases, 9 suspicious cases, and 6 malignant cases. Together, suspicious and malignant cytology cases yielded a sensitivity of 44% and a specificity of 100% for a cytologic diagnosis of cancer. The DNA histogram pattern was D in 24 cases, BD in 9 cases, and A in 16 cases. BD and A patterns were significantly associated with malignancy (P < 0.001). A DH > or = 1% was noted in 22 cases. DH alone had a sensitivity of 62% and a specificity of 91% and was significantly associated with malignancy (P < 0.004). Atypical cytology alone had a false-negative rate of 29%, but in combination with a DH > or = 1%, the false-negative rate decreased to 7%. Additionally, when the authors combined atypical, suspicious, and malignant cytology with a DH > or = 1%, the diagnostic sensitivity increased to 88%, but the specificity decreased to 73%. CONCLUSIONS: Combined cytology and image analysis of bile duct brushing increased diagnostic sensitivity compared with cytology alone. The findings suggest that image analysis may help select patients having atypical cytology who should undergo a more rigorous evaluation for malignancy. A larger prospective study of the usefulness of combined cytology and image analysis of bile duct brushing is warranted. PMID- 11391613 TI - Immunity against solid tumors? PMID- 11391614 TI - Frameshift peptide-derived T-cell epitopes: a source of novel tumor-specific antigens. AB - Microsatellite instability (MSI) caused by defective DNA mismatch repair (MMR) is a hallmark of hereditary nonpolyposis colorectal cancers (HNPCC) but also occurs in about 15% of sporadic tumors. If instability affects microsatellites in coding regions, translational frameshifts lead to truncated proteins often marked by unique frameshift peptide sequences at their C-terminus. Since MSI tumors show enhanced lymphocytic infiltration and our previous analysis identified numerous coding mono- and dinucleotide repeat-bearing candidate genes as targets of genetic instability, we examined the role of frameshift peptides in triggering cellular immune responses. Using peptide pulsed autologous CD40-activated B cells, we have generated cytotoxic T lymphocytes (CTL) that specifically recognize HLA-A2.1-restricted peptides derived from frameshift sequences. Among 16 frameshift peptides predicted from mutations in 8 different genes, 3 peptides conferred specific lysis of target cells exogenously loaded with cognate peptide. One peptide derived from a (-1) frameshift mutation in the TGFbetaIIR gene gave rise to a CTL bulk culture capable of lysing the MSI colorectal cancer cell line HCT116 carrying this frameshift mutation. Given the huge number of human coding microsatellites and assuming only a fraction being mutated and encoding immunologically relevant peptides in MSI tumors, frameshift protein sequences represent a novel subclass of tumor-specific antigens. It is tempting to speculate that a frameshift peptide-directed vaccination approach not only could offer new treatment modalities for existing MSI tumors but also might benefit asymptomatic at-risk individuals in HNPCC families by a prophylactic vaccination strategy. PMID- 11391615 TI - Systematic identification of genes with coding microsatellites mutated in DNA mismatch repair-deficient cancer cells. AB - Microsatellite instability (MSI) caused by deficient DNA mismatch-repair functions is a hallmark of cancers associated with the hereditary nonpolyposis colorectal cancer (HNPCC) syndrome but is also found in about 15% of all sporadic tumors. Most affected microsatellites reside in untranslated intergenic or intronic sequences. However, recently few genes with coding microsatellites were also shown to be mutational targets in MSI-positive cancers and might represent important mutation targets in their pathogenesis. The systematic identification of such genes and the analysis of their mutation frequency in MSI-positive cancers might thus reveal major clues to their functional role in MSI-associated carcinogenesis. We therefore initiated a systematic database search in 33,595 distinctly annotated human genes and identified 17,654 potentially coding mononucleotide repeats (cMNRs) and 2,028 coding dinucleotide repeats (cDNRs), which consist of n > or = 6 and n > or = 4 repeat units, respectively. Expression pattern and mutation frequency of 19 of these genes with the longest repeats were compared between DNA mismatch repair-deficient (MSI(+)) and proficient (MSS) cancer cells. Instability frequencies in these coding microsatellite genes ranged from 10% to 100% in MSI-H tumor cells, whereas MSS cancer cells did not show mutations. RT-PCR analysis further showed that most of the affected genes (10/15) were highly expressed in tumor cells. The approach outlined here identified a new set of genes frequently affected by mutations in MSI-positive tumor cells. It will lead to novel and highly specific diagnostic and therapeutic targets for microsatellite unstable cancers. PMID- 11391616 TI - Cell-transforming activity and estrogenicity of bisphenol-A and 4 of its analogs in mammalian cells. AB - Estrogenicity is an important mechanism in hormonal carcinogenesis but not sufficient to explain the carcinogenic activity of all estrogens. Additional mechanisms, related to genetic alterations, in conjunction with estrogenicity mediated through the estrogen receptor, have been suggested. An environmental estrogen bisphenol-A (BP-A) and its analogs are widespread in our living environment. Because of the potential for human exposure, the possible relationship between carcinogenicity and estrogenicity of these bisphenols was studied using mammalian cells. We quantitatively compared the cell-transforming activity of BP-A and 4 of its analogs (BP-2, BP-3, BP-4 and BP-5) in Syrian hamster embryo (SHE) cells lacking estrogen-receptor expression. The transforming activity determined by the morphological transformation frequencies in SHE cells treated with the bisphenols ranked: BP-4 > BP-5 > BP-3 > BP-A > BP-2. We also compared the estrogenicity of the 5 bisphenols in MCF7 human breast cancer cells as determined by cell proliferation or progesterone receptor (PgR) expression assayed by RT-PCR. When MCF7 cells were treated with the bisphenols, the proliferative potency ranked: BP-A > BP-5 > BP-4 > BP-3 = BP-2. The level of mRNA for PgR in cells treated with the bisphenols was BP-A > BP-5 > BP-4 > BP-3 > BP 2. These indicate that the transforming activity does not correlate with the estrogenicity of the bisphenols, except for BP-2 that has the weakest activity at the both endpoints. In addition, our results suggest that bisphenols with few, if any, transforming and estrogenic activities could be altered by a modification of the chemical structure. Published 2001 Wiley-Liss, Inc. PMID- 11391617 TI - Overexpression of Bcl-2 regulates sodium butyrate- and/or docetaxel-induced apoptosis in human bladder cancer cells both in vitro and in vivo. AB - Sodium butyrate (NaBt), a physiologically occurring short-chain fatty acid, induces differentiation as well as apoptosis in numerous cell types, and this induction is partially regulated by Bcl-2 expression. The objectives of our study were to characterize the in vitro effects of NaBt and/or docetaxel on the growth, cell cycle and apoptosis of human bladder cancer cells, and to determine whether tumor growth in vivo is inhibited by isobutyramide, an orally bioavailable Bt analogue, and/or docetaxel by using Bcl-2-transfected human bladder cancer cell line KoTCC-1/BH and control vector only-transfected cell line KoTCC-1/C. NaBt caused a decrease in growth of both KoTCC-1/C and KoTCC-1/BH cells, however, its growth inhibitory effect was significantly greater in KoTCC-1/C cells. One mM NaBt resulted in G1 cell cycle arrest, accompanied by up-regulation of p21 (waf1/cip1) and down-regulation of cyclin D1 in KoTCC-1/C cells, whereas KoTCC 1/BH showed resistance to G1 cell cycle arrest. An amount of 5 mM NaBt induced apoptosis, accompanied by up-regulation of Bak in KoTCC-1/C cells but failed to induce apoptosis in KoTCC-1/BH cells despite substantial down-regulation of Bcl 2. Oral administration of isobutyramide significantly reduced the KoTCC-1/C tumor volume compared with the KoTCC-1/BH tumor volume in nude mice. Furthermore, docetaxel induced Bcl-2 phosphorylation in KoTCC-1/BH cells and combined treatment with isobutyramide and docetaxel synergistically inhibited the growth of KoTCC-1/BH cells both in vitro and in vivo. These findings suggest that isobutyramide therapy could be a novel therapeutic strategy for patients with bladder cancer if docetaxel is combined according to the Bcl-2 expression status. PMID- 11391618 TI - 8-Cl-cAMP induces cell cycle-specific apoptosis in human cancer cells. AB - 8-Cl-cyclic adenosine monophosphate (8-Cl-cAMP) has been known to induce growth inhibition and differentiation in a variety of cancer cells by differential modulation of protein kinase A isozymes. To understand the anticancer activity of 8-Cl-cAMP further, we investigated the effect of 8-Cl-cAMP on apoptosis in human cancer cells. Most of the tested human cancer cells exhibited apoptosis upon treatment with 8-Cl-cAMP, albeit with different sensitivity. Among them, SH-SY5Y neuroblastoma cells and HL60 leukemic cells showed the most extensive apoptosis. The effect of 8-Cl-cAMP was not reproduced by other cAMP analogues or cAMP elevating agents, showing that the effect of 8-Cl-cAMP was not caused by simple activation of protein kinase A (PKA). However, competition experiments showed that the binding of 8-Cl-cAMP to the cAMP receptor was essential for the induction of apoptosis. After the treatment of 8-Cl-cAMP, cells initially accumulated at the S and G2/M phases of the cell cycle and then apoptosis began to occur among the population of cells at the S/G2/M cell cycle phases, indicating that the 8-Cl-cAMP-induced apoptosis is closely related to cell cycle control. In support of this assumption, 8-Cl-cAMP-induced apoptosis was blocked by concomitant treatment with mimosine, which blocks the cell cycle at early S phase. Interestingly, 8-Cl-cAMP did not induce apoptosis in primary cultured normal cells and non-transformed cell lines, showing that 8-Cl-cAMP-induced apoptosis is specific to transformed cells. Taken together, our results show that the induction of apoptosis is one of the mechanisms through which 8-Cl-cAMP exerts anticancer activity. PMID- 11391619 TI - Granulocyte colony-stimulating factor (G-CSF)-mediated signaling regulates type IV collagenase activity in head and neck cancer cells. AB - Granulocyte colony-stimulating factor (G-CSF), a hematopoietic cytokine, regulates the proliferation and differentiation of granulocytic progenitor cells and functionally activated mature neutrophils. G-CSF also affects nonhematopoietic tumor cells through its binding to the specific receptor (G CSFR) on the cells. The type IV collagenase [matrix metalloproteinase 2 (MMP-2)] is known to play a main role in the process of invasion and metastasis, but its regulation, for example, in expression or in activation, is not clearly understood. In this study, we investigated the role of G-CSF in the regulation of tumor cell invasion and the synthesis of MMP-2. G-CSFs producing the head and neck carcinoma cell line T3M-1 cells with metastatic ability and no G-CSF receptor (G-CSFR) expression were transfected with G-CSFR expression vector. In vitro treatment of G-CSFR-transfectant T3M-1 cells with recombinant G-CSF (rG CSF) significantly augmented their invasive potential in a reconstituted basement membrane (Matrigel) system compared with that of parental cells. Moreover, MMP-2 activity of G-CSFR-transfectant T3M-1 cells was enhanced by the stimulation with rG-CSF, as assessed by gelatin zymography. These results identify G-CSF as a regulator of MMP-2 and cellular invasion. PMID- 11391620 TI - Placental bone morphogenetic protein (PLAB) gene expression in normal, pre malignant and malignant human prostate: relation to tumor development and progression. AB - The second most common target of prostate-cancer metastasis is bone, and the phenomenon of skeletal metastasis represents the incurable stage of disease. Histologically, skeletal metastasis from prostate cancer is distinctive due to its osteoblastic nature. The osteoblastic bone metastasis shows extensive new bone formation, with possible involvement of the soluble growth factors secreted by tumor cells, such as bone morphogenetic proteins (BMPs). In the present study, we analyzed the gene expression of one of the new members of the BMP family, placental bone morphogenetic protein (PLAB). In situ hybridization studies showed high levels of this gene in normal prostate. However, the gene is down-regulated during the progression of cancer at the primary site. The most significant finding was re-expression of the PLAB gene in osseous metastatic lesions. Our results demonstrate that tumor cells, when released from the primary site and after re-growth elsewhere, are capable of re-expressing specific genes that may play a different role at metastatic sites than at the primary site. PMID- 11391621 TI - N-[3,4-dimethoxycinnamoyl]-anthranilic acid (tranilast) inhibits transforming growth factor-beta relesase and reduces migration and invasiveness of human malignant glioma cells. AB - Extensive infiltration of normal brain tissue and suppression of anti-tumor immune surveillance mediated by molecules such as transforming growth factor-beta (TGF-beta) are key biological features that contribute to the malignant phenotype of human gliomas. Tranilast (N-[3,4-dimethoxycinnamoyl]-anthranilic acid) is an anti-allergic compound used clinically to control atopic and fibrotic disorders. These effects are attributed to the suppression of TGF-beta1 synthesis and interference with growth factor-mediated proliferation and migration of fibroblasts and vascular smooth muscle cells. Here, we show that tranilast inhibits DNA synthesis and proliferation of human malignant glioma cells and promotes p21 accumulation in the absence of cytotoxicity. Further, tranilast reduces the release of TGF-beta1 and TGF-beta2 by glioma cells and inhibits migration, chemotactic responses and invasiveness. These effects are not associated with a reduction of alpha(v)beta(3) integrin expression at the cell surface but appear to involve inhibition of matrix metalloproteinase-2 expression and activity. Neither the tranilast-mediated inhibition of proliferation nor the inhibition of migration was counteracted by supplementation with exogenous TGF beta. Finally, tranilast administered orally inhibited the growth of experimental 9L rat gliomas and reduced expression of TGF-beta2 in vivo. We conclude that tranilast might be a useful therapeutic agent for the treatment of human malignant glioma because of a TGF-beta-independent abrogation of the malignant phenotype of proliferation, migration and invasiveness and because of the antagonism of TGF-beta-associated immunosuppression. PMID- 11391622 TI - Expression of multidrug-resistance P-glycoprotein (MDR1) in human brain tumors. AB - Multidrug resistance (MDR) is associated with the expression of P-glycoprotein (P gp), an ATP-dependent transporter which expels anti-cancer drugs from cells. In the present study, MDR1 P-gp was immunodetected by Western blot analysis in 60 human brain tumors, including meningiomas, schwannomas, low-grade gliomas (astrocytomas, pilocytic astrocytomas) and high-grade gliomas (anaplastic astrocytomas, glioblastomas and anaplastic oligodendrogliomas). Most samples from primary tumors expressed P-gp at the same levels as normal brain tissue except for schwannomas, in which levels were reduced by 65%, and meningiomas, in which levels were more than 10-fold higher in 7 of 10 samples. P-gp levels were 70% and 95% lower in brain metastases from melanomas and lung adenocarcinomas, respectively, than in normal brain tissue. These results indicate that the majority of primary brain tumors express MDR1 P-gp and that its high expression levels in meningiomas may be a marker for this type of brain tumor. PMID- 11391623 TI - Human trypsinogen in colorectal cancer. AB - Trypsinogen (TRY), the precursor to the serine protease trypsin, is found in the pancreas and mediates digestive proteolysis in the small intestine. Differential display of cDNAs expressed by human colorectal tumor tissues compared with adjacent normal colonic mucosa identified an isoform of TRY (TRY2) up-regulated in colorectal cancers. Northern blot analysis of RNA isolated from a series of 28 malignant colon tumors and corresponding normal mucosa showed that TRY transcripts were up-regulated 2- to 33-fold in 29% of tumors. Further, TRY mRNA was expressed in 6 colorectal cancer cell lines, with highest levels detected in the metastatic tumor lines SW620 and HT29. Immunostaining for TRY protein expression showed intense immunoreactivity in the supranuclear cytoplasm of colon tumors in 16% of tissue specimens. To evaluate the relative contributions of 2 isoforms of TRY, TRY1 and TRY2, to total TRY mRNA expression, a semi-quantitative multiplex RT-PCR assay was developed. TRY2 mRNA was detected in all 6 colorectal tumor cell lines, whereas TRY1 mRNA was expressed only in the metastatic tumor lines, showing that the high levels of TRY expression in the metastatic tumor lines are likely due to up-regulation of TRY1. Evaluation of TRY1 and TRY2 mRNA expression by multiplex RT-PCR in a series of 20 colon tumor tissues representative of the range of tumor progression showed that TRY2 mRNA was expressed much more commonly than TRY1 mRNA in normal mucosa (26% vs. 6%) as well as in primary tumor tissues (65% vs. 15%). These data demonstrate that TRY2 is the dominant TRY in colon tissue and suggest that up-regulation of TRY1 expression in colon tumors may be associated with a metastatic phenotype. PMID- 11391624 TI - Deletion mapping of chromosome 16q24 in hepatocellular carcinoma in Taiwan and mutational analysis of the 17-beta-HSD gene localized to the region. AB - Human chromosome band 16q24 commonly undergoes loss of heterozygosity (LOH) in human hepatocellular carcinoma (HCC). To further localize the region of deletion on 16q24 and to evaluate the genetic role of 17-beta-HSD, which is near 16q24, in HCC, we examined the pattern of loss of heterozygosity in 88 HCC patients. DNAs from 88 pairs of HCCs and corresponding non-tumor parts were prepared. Loss of heterozygosity on chromosomes 16q24 was investigated by 11 sets of microsatellite markers. Mutation analysis of type II 17-beta-HSD was performed by automatic sequencing. LOH on 16q24 for at least 1 locus was found in 43 of the 88 tumor DNAs (49%). Three non-overlapping regions of frequent LOH were defined in these 43 tumors with partial deletions. The first region was between D16S516 loci and D16S507, encompassed by a 1-cM region, defined by the D16S504. The second region was defined by the 17HSDB2 locus between D16S505 and D16S422, encompassed approximately by a 1-cM region. The third region was between D16S520 and D16S413, defined by D16S3048, encompassed approximately by a 4-cM region. Homozygous deletions of any exons in 17HSDB2 gene were identified in 7 of 27 cases (26%). Automated sequencing analysis of 17HSDB2 failed to demonstrate mutations in any of these specimens. Our data suggest that the 17HSDB2 locus is a frequent target of deletion in HCC but the inactivation of 17HSDB2 may not involve sequence mutations. Furthermore, the presence of the other 2 frequent LOH regions suggest that the putative tumor suppressor genes at these locations might be involved in the development of HCC. PMID- 11391625 TI - Clinical relevance of genomic aberrations in homogeneously treated high-risk stage II/III breast cancer patients. AB - Little is known about the prognostic impact of chromosome aberrations in breast cancer. The aim of our study was to determine whether genomic aberrations of prognostic relevance can be identified in the context of a clinical study using molecular cytogenetics. Paraffin-embedded tumor samples of 44 patients with high risk stage II/III breast cancer were analyzed by comparative genomic hybridization. All patients received identical therapy including dose-escalated chemotherapy followed by peripheral blood stem cell transplantation. The most frequent chromosomal aberrations were gains on chromosome arms 17q (24 cases), 1q (21 cases), 8q (17 cases), 20q (13 cases), 6p (9 cases) as well as losses on chromosome arms 13q (25 cases), 11q (20 cases), 5q (11 cases), 6q (11 cases), 9p (10 cases), 18q (10 cases), 8p (9 cases) and 16q (9 cases). In univariate analysis, the correlation with the clinical outcome revealed a higher risk for patients with tumors exhibiting 13q losses and a reduced risk for tumors exhibiting 16q losses (p = 0.020), 6q losses (p = 0.041) and estrogen-receptor positivity (0.051). In multivariate analysis using the Cox model, only the loss of 16q exhibited borderline significance (p = 0.065). These data show that comparative genomic hybridization can be performed in the context of a clinical trial. In our subgroup of high-risk breast cancer patients, chromosomal aberrations were valuable prognostic parameters. PMID- 11391626 TI - MAGE, BAGE and GAGE gene expression in human rhabdomyosarcomas. AB - MAGE, BAGE and GAGE genes encode tumor-associated antigens that are presented by HLA class I molecules and recognized by CD8(+) cytolytic T lymphocytes. These antigens are currently regarded as promising targets for active, specific tumor immunotherapy because MAGE, BAGE and GAGE genes are expressed in many human cancers of different histotype and are silent in normal tissues, with the exception of spermatogonia and placental cells. MAGE, BAGE and GAGE gene expression has been extensively studied in different tumors of adults but is largely unknown in many forms of pediatric solid cancer. Using RT-PCR, we analyzed MAGE-1, MAGE-2, MAGE-3, MAGE-4, MAGE-6, BAGE, GAGE-1,-2 or -8 and GAGE 3,-4,-5,-6 or -7b gene expression in 31 samples of pediatric rhabdomyosarcoma, the most frequent form of malignant soft tissue tumor in children. MAGE genes were expressed in a substantial proportion of patients (MAGE-1, 38%; MAGE-2, 51%; MAGE-3, 35%; MAGE-4, 22%; MAGE-6, 35%), while expression of BAGE (6%); GAGE-1, GAGE-2 and GAGE-8 (9%); and GAGE-3, GAGE-4, GAGE-5, GAGE-6 and GAGE-7B (16%) was less frequent. Overall, 58% of tumors expressed at least 1 gene, and 35% expressed 3 or more genes simultaneously. Our data suggest that a subset of rhabdomyosarcoma patients could be eligible for active, specific immunotherapy directed against MAGE, BAGE and GAGE antigens. PMID- 11391627 TI - Expression of human tumor-associated antigen RCAS1 in Reed-Sternberg cells in association with Epstein-Barr virus infection: a potential mechanism of immune evasion. AB - RCAS1 (receptor-binding cancer antigen expressed on SiSo cells) is present in neoplastic cells, induces apoptosis of natural killer (NK)/T cells and plays a role in immune evasion. Fas ligand (FasL) is considered to have similar roles. The Epstein-Barr virus (EBV)-encoded latent membrane protein is expressed by malignant Hodgkin and Reed-Sternberg (H&RS) cells of EBV-associated Hodgkin's disease (HD) and considered to be a target of cytotoxic T lymphocytes (CTLs). However, CTL response is inadequate in HD. To determine whether RCAS1 and FasL are expressed in EBV-associated HD and participate in immune evasion, tissues of 20 EBV(-) and 15 EBV(+) HD cases were immunohistochemically stained for RCAS1, FasL and HLA classes I and II, whose deficiencies could explain CTL escape. Lymphocytes surrounding H&RS cells tended to be CD4(+) cells and rarely CD8(+), TIA-1(+) (cytotoxic marker) or NK cells. HLA class I and/or II were expressed in all EBV(+) HD cases, and RCAS1-expressing H&RS cells were found in 14/15 (93%) EBV(+) HD cases but only 8/20 (40%) EBV(-) HD cases (p < 0.05). FasL was detected in 9/15 (60%) and 7/20 (35%) EBV(+) and EBV(-) HD cases, respectively. ssDNA positive (apoptotic) lymphocytes, surrounding H&RS cells, were rarely seen but were present in RCAS1(+) cases (20/22 cases, 91%) rather than negative cases (0/13 cases, 0%) (p < 0.005). Our findings suggest that EBV(+) H&RS cells might evade the host immune response by expressing RCAS1 rather than FasL. PMID- 11391628 TI - Antibody-dependent cell-mediated cytotoxicity can be induced by MUC1 peptide vaccination of breast cancer patients. AB - Human polymorphic epithelial mucin (PEM, MUC1) is a high molecular weight transmembrane glycoprotein expressed on the apical cell surface of glandular epithelium and is over-expressed and hypo-glycosylated in adenocarcinomas. The extracellular part of the molecule consists mainly of a variable number of 20 amino acid repeats that contain cryptic epitopes exposed in malignancy. The objective of our study was to determine whether humanized MUC1 MAbs and Abs induced by vaccination of breast cancer patients with MUC1 peptides can effect an antibody-dependent cell-mediated cytotoxicity (ADCC). An in vitro assay has been set up in which the breast tumor cell line ZR-75-1 is used as target and PBMC of healthy donors as effector cells. Different target and effector cells, as well as various MUC1 MAbs were tested to optimize the efficacy of the in vitro assay. The humanized MAb HuHMFG-1, which recognizes the PDTR sequence in the MUC1 tandem repeat, induced a strong cell-mediated cytotoxicity. Nine MUC1-expressing tumor cell lines, including 3 bone marrow-derived cell lines, as well as 2 MUC1 transfected cell lines were susceptible to different extent to MUC1 Ab-dependent killing. Large variations in the killing capacity of PBMC from healthy donors were found. The NK cells were the essential effector cells for the MUC1 Ab dependent killing. Plasma samples with induced high levels of MUC1 Ab were obtained from breast cancer patients repeatedly immunized with a KLH-conjugated 33-mer or 106-mer MUC1 tandem repeat. Pre- and post-vaccinated plasma samples of these patients were compared in the ADCC assay and it could be clearly demonstrated that the induced MUC1 Abs can effect tumor cell killing. MUC1 Ab dependent cell-mediated tumor cell killing may occur in vivo and the ADCC assay can be applied to monitor MUC1 vaccination trials. PMID- 11391629 TI - Reversal of drug resistance mediated by multidrug resistance protein (MRP) 1 by dual effects of agosterol A on MRP1 function. AB - We previously isolated agosterol A (AG-A) from a marine Spongia sp. and found that it completely reversed colchicine resistance in P-glycoprotein (Pgp)-over expressing KB-C2 cells and vincristine resistance in multidrug-resistance protein (MRP)1-over-expressing CV60 cells. However, a tri-deacetylated derivative of AG-A (IAG-A) showed almost no activity in reversing Pgp- or MRP1-mediated drug resistance. In this study, we examined the mechanisms by which AG-A reverses MRP1 mediated drug resistance by investigating the interaction between agosterols and MRP1 in MRP1-over-expressing human KB carcinoma (KB/MRP) cells. [3H]-Leukotriene C4 (LTC4), [3H]-2,4-dinitrophenyl-S-glutathione uptake into membrane vesicles prepared from KB/MRP cells and intracellular [3H]-vincristine accumulation and efflux in KB/MRP cells were measured with or without AG-A and/or inactive IAG-A. AG-A reduced MRP1-mediated [3H]-LTC4 transport in a dose-dependent manner, but IAG-A did not. Inhibition by AG-A was competitive, with a K(i) value of 31 microM. AG-A at 10 microM enhanced the accumulation of [3H]-vincristine in KB/MRP cells to the level of that in control cells in the absence of the agent. Likewise, ATP-dependent efflux of [3H]-vincristine from KB/MRP cells was enhanced compared with KB-3-1 cells and inhibited by AG-A. In addition, AG-A reduced intracellular levels of glutathione, a compound required for MRP1-mediated transport of some anti-cancer drugs. These findings suggest that AG-A reverses MRP1-mediated drug resistance by directly inhibiting the capacity of MRP1 to transport drugs. In addition, the capacity of AG-A to reduce cellular glutathione levels may contribute to the modulating activity of MRP1. PMID- 11391630 TI - Determining MDR1/P-glycoprotein expression in breast cancer. AB - The mechanism of chemotherapy resistance in breast cancer is unresolved. MDR1/P glycoprotein (P-Gp) over-expression confers multidrug resistance in vitro and might play a role in clinical breast cancer. Studies using clinical samples have yielded conflicting results. MDR1/P-Gp mRNA expression was determined relative to the expression in normal human liver using TaqMan real-time RT-PCR (corrected for expression of the housekeeping gene PBGD). Immunohistochemistry (IHC) was performed with monoclonal antibodies against P-Gp (JSB1, C219). The positive control was SW1573/2R160, the intermediate control SW1573 and the negative control GLC4/ADR. We assayed 9 breast-cancer cell lines by RT-PCR and IHC, 52 carcinoma samples by RT-PCR and 168 samples by IHC. SW1573/2R160 contained high levels of MDR1/P-Gp mRNA (1.0, equal to liver) and showed strong membranous staining. Expression of MDR1/P-Gp mRNA in SW1573 (0.05) and GLC4/ADR (3.2 x 10( 5)) was not detectable by IHC. Very low levels of MDR1/P-Gp mRNA were measured in breast-cancer cell lines (mean 3.1 x 10(-4), range 1 to 12 x 10(-4)), but P-Gp was not detected by IHC. In 25 specimens from chemotherapy-naive patients, MDR1/P Gp mRNA levels varied from 1 to 11 x 10(-2) (mean 3.9 x 10(-2)). In sections of 80 chemotherapy-naive tumors, no membrane-bound staining was observed in the tumor cells. Tumors of 27 anthracycline-treated patients had comparable MDR1/P-Gp mRNA expression levels (mean 5.4 x 10(-2)). P-Gp was undetectable in 88 tumor samples of patients who had received anthracycline-based chemotherapy. In breast cancer, MDR1/P-Gp mRNA is low or absent and P-Gp levels in cancer cells are too low to detect by IHC. Chemotherapy exposure does not result in detectable MDR1/P Gp over-expression. PMID- 11391631 TI - Appropriate subcellular localisation of prodrug-activating enzymes has important consequences for suicide gene therapy. AB - Escherichia coli B nitroreductase (NR) has been expressed stably in MDA-MB-361 human breast adenocarcinoma cells either as the wild-type protein (wtNR), which is distributed evenly between the cytoplasmic and nuclear compartments, or targeted to the mitochondrion (mtNR). Whereas bacterial NR is active as a dimer, a proportion of wtNR is monomeric. In contrast, mtNR is mostly dimeric, suggesting that it adopts a more stable, native conformation. Despite this, when tested in gene-directed enzyme prodrug therapy cell cytotoxicity studies, cells expressing wtNR or mtNR had similar sensitivity to the prodrug CB1954 and mounted similar bystander killing effects. Furthermore, when short prodrug exposures were given, wtNR was more efficient at killing cells than mtNR. These data demonstrate that the site of enzyme expression and prodrug activation is an important variable that requires consideration in suicide gene therapy approaches. PMID- 11391632 TI - Glucosylceramide synthase inhibition enhances vincristine-induced cytotoxicity. AB - As a strategy to enhance tumor cell sensitivity to vincristine, we tested whether modulation of sphingolipid metabolism would alter vincristine cytotoxicity since this is linked to accumulation of the intermediate metabolite, ceramide. We blocked ceramide metabolism in a series of variably vincristine-resistant cell lines derived from CCRF-CEM leukemia cells using an inhibitor of glucosylceramide synthase, DL-threo-phenyl-2-hexadecanoylamino-3-pyrrolidino-1-propanol (PPPP). PPPP alone (1.0 microM), while nearly completely blocking glucosylceramide synthesis, was not toxic and did not increase cellular ceramide levels. Vincristine alone was toxic, caused apoptosis or programmed cell death (PCD) and caused an elevation in ceramide levels. Strikingly, the combination of PPPP and vincristine resulted in a further increase, over that of vincristine alone, of (i) cellular ceramide concentration, (ii) cytotoxicity associated with PCD and (iii) G2/M cell-cycle arrest. PPPP had no effect on P-glycoprotein expression or function. We conclude that vincristine cytotoxicity occurs in part through a ceramide-dependent mechanism, resulting in both G2/M block as well as PCD, and that the blockade of glucosylceramide synthase, in itself not toxic, causes augmented accumulation of ceramide resulting from vincristine exposure, which in turn maximizes ceramide-dependent, vincristine-induced cytotoxicity. Inhibition of glucosylceramide synthesis may be a means of circumventing drug resistance by enhancing signaling through a cell-death pathway. PMID- 11391633 TI - Use of paclitaxel in patients with pre-existing cardiomyopathy: a review of our experience. AB - Cardiac toxicity is frequently the indication for discontinuation of an anthracycline in patients with tumors which remain anthracycline-sensitive. During the 1990s, the most frequently used second-line agents at the Yale Cancer Center (YCC) were the taxanes. The goal of this retrospective analysis was to determine the effect of paclitaxel on cardiac function in patients with cardiomyopathy. YCC outpatient clinic pharmacy order forms were used to identify all patients who had received paclitaxel between December 1995 and November 1997. The clinic records of those patients with a left ventricular ejection fraction (LVEF) of < or = 50% were reviewed to determine the temporal relation between the decreased LVEF and paclitaxel therapy. In addition, clinic records were examined for evidence of prior doxorubicin therapy and history of prior cardiac disease. Between December 1995 and November 1997, 225 patients were treated with paclitaxel in the YCC outpatient clinic. Nine patients had LVEF < or = 50% (mean 37%) prior to initiation of paclitaxel therapy. Six of these patients had equilibrium radionuclide angiocardiographic (ERNA) scans following completion of paclitaxel. In these 6 patients, the mean change in LVEF was +6% (range -3% to +29%). Four patients had improved LVEF following paclitaxel (mean 11%, range 2% to 29%), while 2 patients experienced a decrease in LVEF following paclitaxel treatment (mean 2.5%). The 3 patients who did not have ERNA scans following paclitaxel therapy had no clinical evidence of congestive heart failure. Our experience confirms the results of prior studies that paclitaxel can be safely administered in patients with underlying cardiac dysfunction. PMID- 11391634 TI - Population-based cancer survival in Singapore, 1968 to 1992: an overview. AB - The Singapore Cancer Registry has provided comprehensive population-based incidence data since 1968. This paper describes the population-based survival analysis of the registry data. All invasive primary cancers diagnosed from January 1, 1968 to December 31, 1992 were passively followed up until December 31, 1997. Only 5.8% were lost to follow-up. Cumulative and observed survival rates were calculated using Hakulinen's method. Overall 5-year relative survival rates have increased dramatically over the 25-year period in both genders. Significant increases are seen with nasopharynx, stomach and colo-rectum cancers, non-Hodgkin's lymphoma, leukemias and cancers of the testis, cervix, ovaries and breast. When compared with the Surveillance, Epidemiology and End Results (SEER) rates in the United States, the 5-year relative survival rates in Singapore are generally lower. However, the rate of change between the two countries is fairly similar. On the average, the rates are 10 to 15 years behind the SEER rates and 5 to 10 years behind Finland, Switzerland and Japan, but they are close to the UK rates. The age-standardized 5-year survival rate for Singapore is higher for most sites compared with other developing countries like Qidong (China), Madras (India), Bombay (India) and Chiang Mai (Thailand). The 25-year trend in cancer survival in Singapore showed two extreme groups: those showing no change and those showing significant improvements. Reducing the incidence of cancers belonging to the first group remains the only viable mode of cancer control. For cancers in the second group, improvement in survival is due to a combination of successful early detection measures and effective treatment services in Singapore. PMID- 11391635 TI - Family history of cancer and risk of esophageal and gastric cancers in the United States. AB - The worldwide rates for histology- and subsite-specific types of esophageal and gastric cancer reveal strikingly divergent patterns. The contribution of environmental and genetic factors has been explored in several high-incidence areas, but data on genetic influences are scarce for Western countries. Using data from a multicenter, population-based, case-control study on 1,143 cases and 695 controls in the United States, we evaluated whether a family history of digestive or other cancers was associated with an increased risk of esophageal adenocarcinoma (n = 293), esophageal squamous cell carcinoma (n = 221), gastric cardia adenocarcinoma (n = 261) or non-cardia gastric adenocarcinoma (n = 368). After adjusting for other risk factors, individuals reporting a family history of digestive cancers experienced no increased risk of either type of esophageal cancer but they were prone to adenocarcinomas of the gastric cardia [odds ratio (OR) = 1.34, 95% confidence interval (CI) 0.91-1.97] and non-cardia segments (OR =1.46, 95% CI 1.03-2.08). This familial tendency, particularly for non-cardia gastric tumors, was largely explained by an association with family history of stomach cancer (OR = 2.52, 95% CI 1.50-4.23). In addition, family history of breast cancer was associated with increased risks of esophageal adenocarcinoma (OR = 1.74, 95% CI 1.07-2.83) and non-cardia gastric adenocarcinoma (OR = 1.76, 95% CI 1.09-2.82). Also seen were non-significant familial associations of esophageal squamous-cell cancer with prostate cancer as well as non-cardia gastric cancer with leukemia and brain tumors, though these relationships must be interpreted with caution. Our data point to the role of familial susceptibility to gastric cancer, but not to any form of esophageal cancer, in the United States. PMID- 11391636 TI - Postnatal development of corticospinal axon terminal morphology in the cat. AB - The corticospinal system undergoes important postnatal development, leading to the mature topography and specificity of connections. The purpose of this study was to determine the time-course of development of corticospinal axonal branching and varicosity density within the cervical gray matter. Corticospinal neurons were labeled after small injections of the anterograde tracer biotinylated dextran amine into the primary motor cortex of cats. Tracer injection and transport times were adjusted to examine labeling at 25, 35, 55, and 75 days and in adults. We measured the numbers and lengths of nonreconstructed terminal and preterminal branches and the numbers and locations of axon varicosities. We found significant age-dependent increases in all morphologic measures. At 25 days, corticospinal axon branching was sparse, with only a few scattered varicosities. By day 35, the mean number of branches, varicosities per branch, and varicosity density increased. Several morphologic measures did not increase between day 35 and 55, but further changes occurred between 55 days and maturity. Beginning around day 55, there was extensive development of small terminal axon branches with high densities of varicosities. We also found, by using spatial point analysis, that there was an age-dependent increase in varicosity clustering. Our results show for the first time that terminal and preterminal corticospinal axon branches increase in complexity during a protracted early postnatal period. This developmental period extended beyond the early postnatal period of activity dependent refinement of the topography of terminations. Comparison with the time course of maturation of the cortical motor representation revealed development of substantial, albeit incomplete, branching and varicosity density of CS axons before cortical motor circuits effectively drive their spinal targets. PMID- 11391637 TI - High-affinity neurotensin receptors in the rat nucleus accumbens: subcellular targeting and relation to endogenous ligand. AB - Neurotensin is present in selective mesolimbic dopaminergic projections to the nucleus accumbens (NAc) shell but also is synthesized locally in this region and in the motor-associated NAc core. We examined the electron microscopic immunolabeling of the high-affinity neurotensin receptor (NTR) and neurotensin in these subdivisions of rat NAc to determine the sites for receptor activation and potential regional differences in distribution. Throughout the NAc, NTR immunoreactivity was localized discretely within both neurons and glia. NTR labeled neuronal profiles were mainly axons and axon terminals with diverse synaptic structures, which resembled dopaminergic and glutamatergic afferents, as well as collaterals of inhibitory projection neurons. These terminals had a significantly higher numerical density in the NAc core than in the shell but were prevalent in both regions, suggesting involvement in both motor and limbic functions. In each region, neurotensin was detected in a few NTR-immunoreactive axon terminals and in terminals that formed symmetric, inhibitory type synapses with NTR-labeled somata and dendrites. The NTR labeling, however, was not seen within these synapses and, instead, was localized to segments of dendritic and glial plasma membranes often near excitatory type synapses. Neuronal NTR immunoreactivity also was associated with cytoplasmic tubulovesicles and nuclear membranes. Our results suggests that, in the NAc shell and core, NTR is targeted mainly to presynaptic sites, playing a role in the regulated secretion and/or retrograde signaling in diverse, neurotransmitter-specific neurons. The findings also support a volume mode of neurotensin actions, specifically affecting excitatory transmission through activation of not only axonal but also dendritic and glial NTR. PMID- 11391638 TI - Selective localization of high concentrations of F-actin in subpopulations of dendritic spines in rat central nervous system: a three-dimensional electron microscopic study. AB - Dendritic spines differ considerably in their size, shape, and internal organization between brain regions. We examined the actin cytoskeleton in dendritic spines in hippocampus (areas CA1, CA3, and dentate gyrus), neostriatum, and cerebellum at both light and electron microscopic levels by using a novel high-resolution photoconversion method based in the high affinity of phalloidin for filamentous (F)-actin. In all brain regions, labeling was strongest in the heads of dendritic spines, diminishing in the spine neck. The number of labeled spines varied by region. Compared with the cerebellar molecular layer and area CA3, where nearly every dendritic spine was labeled, less than half the spines were labeled in CA1, dentate gyrus, and neostriatum. Serial section reconstructions of spines in these areas indicated that phalloidin labeling was restricted to the largest and most morphologically diverse dendritic spines. The resolution of the photoconversion technique allowed us to examine the localization and organization of actin filaments in the spine. The most intense staining for actin was found in the postsynaptic density and associated with the spines internal membrane system. In mushroom-shaped spines, F-actin staining was particularly strong between the lamellae of the spine apparatus. Three dimensional reconstruction of labeled spines by using electron tomography showed that the labeled dense material was in continuity with the postsynaptic density. These results highlight differences in the actin cytoskeleton between different spine populations and provide novel information on the organization of the actin cytoskeleton in vivo. PMID- 11391639 TI - Development of cholinergic terminals around rat spinal motor neurons and their potential relationship to developmental cell death. AB - Neuron death seems to be regulated mainly by postsynaptic target cells. In chicks, nicotinic antagonists such as alpha-bungarotoxin (alphaBT) can prevent normal cell death of somatic motor neurons (SMNs). For this effect, however, alphaBT could be acting at peripheral neuromuscular junctions and/or central cholinergic synapses. To investigate this issue, we first studied the development of cholinergic terminals in the rat spinal cord by using vesicular acetylcholine transporter immunocytochemistry. Labeled terminals were seen in the ventral horn as early as embryonic day 15 (E15), the beginning of the cell death period. Thus, central cholinergic synapses form at the correct time and place to be able to influence SMN death. We next added alphaBT to organotypic, spinal slice cultures made at E15. After 5 days in vitro, the number of SMNs in treated cultures was substantially greater than in control cultures, indicating that alphaBT can reduce SMN cell death in rats as it does in chicks. Moreover, peripheral target removal led to extensive loss of SMNs, and such a loss occurred even in the presence of alphaBT, indicating the necessity of peripheral target for the alphaBT effect. Finally, to determine whether central cholinergic terminals also may be involved in SMN death, we delayed the alphaBT treatment until after central cholinergic terminals had disappeared from the slice cultures. The increased number of surviving SMNs, even in the absence of central terminals, argued that alphaBT acts at peripheral, not central, cholinergic synapses to rescue SMNs from developmental cell death. PMID- 11391640 TI - Polarized distribution of alpha5 integrin in dendrites of hippocampal and cortical neurons. AB - The distribution of immunoreactivity for the alpha5 subunit of the fibronectin receptor was evaluated in adult rat brain with particular interest in the cellular localization of immunostaining in the hippocampal formation and neocortex. Beyond localization to neuronal perikarya and short dendritic fragments within most brain areas, alpha5 immunoreactivity (-ir) was particularly dense within primary apical dendrites of pyramidal cells in both hippocampus and neocortex and within the dendritic arbors of cerebellar Purkinje cells. In hippocampal and cortical pyramidal cells, immunostaining was clearly polarized: alpha5-ir was not detectable in basal dendrites in hippocampal neurons and was limited to proximal arbors or absent from basal dendrites in pyramidal cells in superficial and deep layers of neocortex. Beyond this, alpha5-ir was distributed within the dendritic ramifications of the dentate gyrus granule cells and within perikarya and dendrites of occasional nonpyramidal neurons. Developmental studies demonstrated that, in both hippocampus and neocortex, alpha5-ir appears first within perikarya and is distributed to dendrites during the second postnatal week. These results are in accord with the broad hypothesis that integrins contribute to apical-basal differences in dendrites and that the integrin fibronectin (alpha5beta1) receptor, in particular, contributes to some late developing features of dendritic structure or function. PMID- 11391641 TI - Immunohistochemical localization of DARPP-32 in the brain of the lizard, Gekko gecko: co-occurrence with tyrosine hydroxylase. AB - To assess the relationship between dopaminergic neuronal structures and dopaminoceptive structures in a reptile, single and double immunohistochemical procedures with antibodies directed against DARPP-32 (dopamine- and cAMP regulated phosphoprotein with an apparent molecular mass of 32,000 daltons),a phosphoprotein related to the dopamine D(1)-receptor, and tyrosine hydroxylase (TH) were applied to the brain of the lizard, Gekko gecko. The DARPP-32 antibody yielded a well-differentiated pattern of staining in the brain of Gekko. In general, areas that are densely innervated by TH-immunoreactive, putative dopaminergic fibers, such as the nucleus accumbens, striatum, dorsal ventricular ridge, and amygdaloid complex, display strong immunoreactivity for DARPP-32 in somata and neuropil. Distinct cellular DARPP-32 immunoreactivity was also found in the lateral cortex, ventral hypothalamus, habenula, central nucleus of the torus semicircularis, midbrain tectum, parvicellular isthmic nucleus, raphe nuclei, caudal rhombencephalic tegmentum, and spinal cord. Striatal projections to the midbrain and their target, i.e., the substantia nigra pars reticulata, were found to be strongly immunoreactive. Double immunofluorescence staining revealed that dopaminergic cells generally do not stain for DARPP-32, except for cells in the ventral hypothalamus and at caudal rhombencephalic levels. In conclusion, the distribution of DARPP-32 in the brain of the lizard Gekko gecko largely resembles the pattern observed in birds and mammals, at least as far as basal ganglia structures are concerned. On the other hand, there are several specific features of DARPP-32 distribution in the gekkonid brain that deserve further attention, such as cellular colocalization of DARPP-32 and TH immunoreactivity in hypothalamic and caudal rhombencephalic areas, and cellular DARPP-32 immunoreactivity in the tectum and central nucleus of the torus semicircularis of the midbrain, the superior and inferior raphe nuclei, and the spinal cord. PMID- 11391642 TI - Immunocytochemical localization of opsin, visual arrestin, myosin III, and calmodulin in Limulus lateral eye retinular cells and ventral photoreceptors. AB - The photoreceptors of the horseshoe crab Limulus polyphemus are classical preparations for studies of the photoresponse and its modulation by circadian clocks. An extensive literature details their physiology and ultrastructure, but relatively little is known about their biochemical organization largely because of a lack of antibodies specific for Limulus photoreceptor proteins. We developed antibodies directed against Limulus opsin, visual arrestin, and myosin III, and we have used them to examine the distributions of these proteins in the Limulus visual system. We also used a commercial antibody to examine the distribution of calmodulin in Limulus photoreceptors. Fixed frozen sections of lateral eye were examined with conventional fluorescence microscopy; ventral photoreceptors were studied with confocal microscopy. Opsin, visual arrestin, myosin III, and calmodulin are all concentrated at the photosensitive rhabdomeral membrane, which is consistent with their participation in the photoresponse. Opsin and visual arrestin, but not myosin III or calmodulin, are also concentrated in extra rhabdomeral vesicles thought to contain internalized rhabdomeral membrane. In addition, visual arrestin and myosin III were found widely distributed in the cytosol of photoreceptors, suggesting that they have functions in addition to their roles in phototransduction. Our results both clarify and raise new questions about the functions of opsin, visual arrestin, myosin III, and calmodulin in photoreceptors and set the stage for future studies of the impact of light and clock signals on the structure and function of photoreceptors. PMID- 11391643 TI - Changes in synaptic inputs to sympathetic preganglionic neurons after spinal cord injury. AB - Spinal cord injury (SCI) leads to plastic changes in organization that impact significantly on central nervous control of arterial pressure. SCI causes hypotension and autonomic dysreflexia, an episodic hypertension induced by spinal reflexes. Sympathetic preganglionic neurons (SPNs) respond to SCI by retracting and then regrowing their dendrites within 2 weeks of injury. We examined changes in synaptic input to SPNs during this time by comparing the density and amino acid content of synaptic input to choline acetyltransferase (ChAT)-immunoreactive SPNs in the eighth thoracic spinal cord segment (T8) in unoperated rats and in rats at 3 days or at 14 days after spinal cord transection at T4. Postembedding immunogold labeling demonstrated immunoreactivity for glutamate or gamma aminobutyric acid (GABA) within presynaptic profiles. We counted the number of presynaptic inputs to measured lengths of SPN somatic and dendritic membrane and identified the amino acid in each input. We also assessed gross changes in the morphology of SPNs using retrograde labeling with cholera toxin B and light microscopy to determine the structural changes that were present at the time of evaluation of synaptic density and amino acid content. At 3 days after SCI, we found that retrogradely labeled SPNs had shrunken somata and greatly shortened dendrites. Synaptic density (inputs per 10-microm membrane) decreased on ChAT immunoreactive somata by 34% but increased on dendrites by 66%. Almost half of the inputs to SPNs lacked amino acids. By 14 days, the density of synaptic inputs to dendrites and somata decreased by 50% and 70%, respectively, concurrent with dendrite regrowth. The proportion of glutamatergic inputs to SPNs in spinal cord transected rats ( approximately 40%) was less than that in unoperated rats, whereas the GABAergic proportion (60-68%) increased. In summary, SPNs participate in vasomotor control after SCI despite profound denervation. An altered balance of excitatory and inhibitory inputs may explain injury-induced hypotension. PMID- 11391644 TI - Effects of age on the thickness of myelin sheaths in monkey primary visual cortex. AB - The effect of age on myelin sheath thickness was determined by an electron microscopic examination of cross sections of the vertical bundles of nerve fibers that pass through primary visual cortex of the rhesus monkey. The tissue was taken from the cortices of young (4-9 years of age) and old (over 24 years of age) monkeys, and the sections were taken at the level of layer 4Cbeta. From the electron photomicrographs, the diameters of axons and the numbers of lamellae in their myelin sheaths were determined. No change was found in the diameters of axons with age, although the mean numbers of myelin lamellae in the sheaths increased from 5.6 in the young monkeys to 7.0 in the old monkeys. Much of this increase in mean thickness was due to the fact that, in the old monkeys, thick myelin sheaths with more than ten lamellae are more common than in the young monkeys. While this increase in the thickness of myelin sheaths is occurring in old monkeys, there are also age-related changes in some of the sheaths. Consequently, it seems that, with age, there is some degeneration of myelin but, at the same time, a continued production of lamellae. PMID- 11391645 TI - Daily variation in the distribution of glycogen phosphorylase in the suprachiasmatic nucleus of Syrian hamsters. AB - Dynamic changes in astrocytic processes in the Syrian hamster suprachiasmatic nucleus (SCN) have been reported with maximal process extension in the light phase and maximal process retraction in the dark phase of a daily light:dark cycle. In the present study, we asked whether dynamic changes occur in the distribution of an astrocytic metabolic marker, glycogen phosphorylase (GP), using a histochemical assay to reveal the distribution of both active and total GP, in the hamster SCN. Changes in glial acidic fibrillary protein (GFAP) immunoreactivity also were assessed using a relative optical density measure (ROD). We observed changes in the localization and distribution of GP both in the SCN and in the paraventricular nucleus of the hypothalamus (PVN) as a function of time of day. In the light phase, there were concentrated, large, dot-like deposits of GP throughout the SCN and PVN on an empty background. In the dark phase, diffuse, small, granular particles were seen throughout both nuclei. Selectively, in the dark-phase SCN, these granular particles formed a rim of intense GP reactivity on the lateral, ventral, posterior, and medial borders. Significantly higher levels of GP reactivity were seen in anterior sections of the medial optic chiasm in the light phase. GFAP-immunoreactive astrocytic processes had higher ROD levels in the dark phase. In conclusion, the astrocytic metabolic marker, GP, exhibits a significant daily variation in localization in both the SCN and the PVN that correlates with dynamic changes in the distribution of astrocytic processes in the SCN. Increased GP activity also occurs in astrocytes among optic fibers subjacent to the SCN during light input. PMID- 11391646 TI - Calcium transients in subcompartments of the leech Retzius neuron as induced by single action potentials. AB - Regional Ca(2+) influx into neurons plays an essential role for fast signal processing, yet it is little understood. We have investigated intracellular Ca(2+) transients induced by a single action potential (AP) in Retzius neurons in situ of isolated ganglia of the leech Hirudo medicinalis using confocal laser scanning microscopy in the cell body, in different axonal branches, and in dendrites. In the cell body, a single AP induced a Ca(2+) transient in submembrane regions, while in central regions no fluorescence change was detected. Burst activity evoked a much larger Ca(2+) influx, which elicited Ca(2+) signals in central somatic regions, including the cell nucleus. A single AP induced a Ca(2+) transient in distal branches of the axon and in dendrites that was significantly larger than in the proximal axon and in the cell body (p <.05), and the recovery of the Ca(2+) transient was significantly faster in axonal branches than in dendrites (p <.01). The AP-induced Ca(2+) transient was inhibited by Co(2+) (2 mM). The P/Q-type Ca(2+) channel blocker omega-agatoxin TK (500 nM) and the L-type Ca(2+) channel blocker nifedipine (20 microM) had no effect on the Ca(2+) transient, whereas the L-type Ca(2+) channel blocker methoxyverapamil (D600, 0.5-1 mM) irreversibly reduced the Ca(2+) transient by 37% in axons and by 42% in dendrites. Depletion of intracellular Ca(2+) stores following inhibition of endoplasmic Ca(2+)-ATPases by cyclopiazonic acid (10 microM) decreased the AP-induced Ca(2+) transient in the dendrites by 21% (p <.01), but not in axons, and increased the Ca(2+) recovery time constant (tau) in the axonal branches by 129% (p <.01), but not in dendrites. The results indicate that an AP evokes a voltage-gated Ca(2+) influx into all subcompartments of the Retzius neuron, where it produces a Ca(2+) signal of different size and/or kinetics. This may contribute to the modulation of electrical excitation and propagation of APs, and to different modes of synaptic and nonsynaptic processes. PMID- 11391647 TI - Pigment-dispersing factor in the locust abdominal ganglia may have roles as circulating neurohormone and central neuromodulator. AB - Pigment-dispersing factor (PDF) is a neuropeptide that has been indicated as a likely output signal from the circadian clock neurons in the brain of Drosophila. In addition to these brain neurons, there are PDF-immunoreactive (PDFI) neurons in the abdominal ganglia of Drosophila and other insects; the function of these neurons is not known. We have analyzed PDFI neurons in the abdominal ganglia of the locust Locusta migratoria. These PDFI neurons can first be detected at about 45% embryonic development and have an adult appearance at about 80%. In each of the abdominal ganglia (A3-A7) there is one pair of lateral PDFI neurons and in each of the A5-A7 ganglia there is additionally a pair of median neurons. The lateral neurons supply varicose branches to neurohemal areas of the lateral heart nerves and perisympathetic organs, whereas the median cells form processes in the terminal abdominal ganglion and supply terminals on the hindgut. Because PDF does not influence hindgut contractility, it is possible that also these median neurons release PDF into the circulation. Release from one or both the PDFI neuron types was confirmed by measurements of PDF-immunoreactivity in hemolymph by enzyme immunoassay. PDF applied to the terminal abdominal ganglion triggers firing of action potentials in motoneurons with axons in the genital nerves of males and the 8th ventral nerve of females. Because this action is blocked in calcium-free saline, it is likely that PDF acts via interneurons. Thus, PDF seems to have a modulatory role in central neuronal circuits of the terminal abdominal ganglion that control muscles of genital organs. PMID- 11391648 TI - Sodium channel distribution on uninnervated and innervated embryonic skeletal myotubes. AB - Acetylcholine receptor (AChR) and sodium (Na(+)) channel distributions within the membrane of mature vertebrate skeletal muscle fibers maximize the probability of successful neuromuscular transmission and subsequent action potential propagation. AChRs have been studied intensively as a model for understanding the development and regulation of ion channel distribution within the postsynaptic membrane. Na(+) channel distributions have received less attention, although there is evidence that the temporal accumulation of Na(+) channels at developing neuromuscular junctions (NMJs) may differ between species. Even less is known about the development of extrajunctional Na(+) channel distributions. To further our understanding of Na(+) channel distributions within junctional and extrajunctional membranes, we used a novel voltage-clamp method and fluorescent probes to map Na(+) channels on embryonic chick muscle fibers as they developed in vitro and in vivo. Na(+) current densities on uninnervated myotubes were approximately one-tenth the density found within extrajunctional regions of mature fibers, and showed several-fold variations that could not be explained by a random scattering of single channels. Regions of high current density were not correlated with cellular landmarks such as AChR clusters or myonuclei. Under coculture conditions, AChRs rapidly concentrated at developing synapses, while Na(+) channels did not show a significant increase over the 7 day coculture period. In vivo investigations supported a significant temporal separation between Na(+) channel and AChR aggregation at the developing NMJ. These data suggest that extrajunctional Na(+) channels cluster together in a neuronally independent manner and concentrate at the developing avian NMJ much later than AChRs. PMID- 11391649 TI - Blockade of the central generator of locomotor rhythm by noncompetitive NMDA receptor antagonists in Drosophila larvae. AB - The noncompetitive antagonists of the vertebrate N-methyl-D-aspartate (NMDA) receptor dizocilpine (MK 801) and phencyclidine (PCP), delivered in food, were found to induce a marked and reversible inhibition of locomotor activity in Drosophila melanogaster larvae. To determine the site of action of these antagonists, we used an in vitro preparation of the Drosophila third-instar larva, preserving the central nervous system and segmental nerves with their connections to muscle fibers of the body wall. Intracellular recordings were made from ventral muscle fibers 6 and 7 in the abdominal segments. In most larvae, long-lasting (>1 h) spontaneous rhythmic motor activities were recorded in the absence of pharmacological activation. After sectioning of the connections between the brain and abdominal ganglia, the rhythm disappeared, but it could be partially restored by perfusing the muscarinic agonist oxotremorine, indicating that the activity was generated in the ventral nerve cord. MK 801 and PCP rapidly and efficiently inhibited the locomotor rhythm in a dose-dependent manner, the rhythm being totally blocked in 2 min with doses over 0.1 mg/mL. In contrast, more hydrophilic competitive NMDA antagonists had no effect on the motor rhythm in this preparation. MK 801 did not affect neuromuscular glutamatergic transmission at similar doses, as demonstrated by monitoring the responses elicited by electrical stimulation of the motor nerve or pressure applied glutamate. The presence of oxotremorine did not prevent the blocking effect of MK 801. These results show that MK 801 and PCP specifically inhibit centrally generated rhythmic activity in Drosophila, and suggest a possible role for NMDA like receptors in locomotor rhythm control in the insect CNS. PMID- 11391650 TI - 22q13 deletion syndrome. AB - We have recently collected clinical information on 37 individuals with deletion of 22q13 and compared the features of these individuals with 24 previously reported cases. The features most frequently associated with this deletion are global developmental delay, generalized hypotonia, absent or severely delayed speech, and normal to advanced growth. Minor anomalies include dolicocephaly, abnormal ears, ptosis, dysplastic toenails, and relatively large hands. As with many terminal deletions involving pale G-band regions, the deletion can be extremely subtle and can go undetected on routine cytogenetic analysis. In fact, 32% of the individuals in our study had previous chromosome analyses that failed to detect the deletion. Eight of 37 individuals had deletion of 22q13 secondary to an unbalanced chromosome translocation. In the newborn, this deletion should be considered in cases of hypotonia for which other common causes have been excluded. In the older child, this syndrome should be suspected in individuals with normal growth, profound developmental delay, absent or delayed speech, and minor dysmorphic features. We recommend high-resolution chromosome analysis and fluorescence in situ hybridization studies, or molecular analysis to exclude this diagnosis. PMID- 11391651 TI - Interstitial duplication of the short arm of chromosome 1 in a newborn with congenital heart disease and multiple malformations. AB - Interstitial duplications of chromosomes 1p are rare, with only 14 cases previously reported in the literature, and those have not revealed a unique syndrome. The phenotypes include multiple congenital abnormalities and both intra and extra-uterine growth retardation. In general, the patients do poorly and do not survive beyond the age of several months. We report a newborn male with karyotype 46, XY, inv dup(1)(qter--> p34.3::p34.3-->p32.3::34.3-->pter) with multiple congenital abnormalities including congenital heart disease and co existing portal and pulmonary hypertension. The chromosome 1 origin of the extra material was confirmed with fluorescent in situ hybridization (FISH). Review of the GDB [Human Genome Database, 1990] reveals that the duplicated region includes the locus EDN2 that encodes endothelin-1, a potent vasoconstrictor, making genetic overdosage of this protein a likely etiology of the pulmonary hypertension. The diffuse abnormalities show effects in multiple cell lines and suggest that this region of chromosome 1p could be involved in determining cell migration and/or differentiation during organogenesis. PMID- 11391652 TI - Bends in human mitotic metaphase chromosomes revisited: 15q11-13 is the most frequent non-random autosomal bend in blood cultures. AB - We have investigated the preferential bending of some chromosome sites in blood cultures from normal and chromosomally abnormal subjects. A total of 2,262 centromeric and 2,718 non-centromeric bends were recorded, and 69 non-centromeric sites were found not to bend at random. 15q11-13 bending was found to be the most frequent non-random autosomal bend. Bends on chromosomes may be remnants of a folded chromosome state in the nucleus, and may facilitate the preferential involvement of some chromosomal bands in structural reorganizations such as the isoacentric fragments, or contribute to the high frequency of interstitial deletions and isodicentric inversion duplications involving the 15q11-13 region. PMID- 11391653 TI - Genetic mapping of a novel X-linked recessive colobomatous microphthalmia. AB - Colobomatous microphthalmia is a common ocular malformation with a heterogeneous phenotype. The majority of cases without associated systemic abnormalities have an autosomal dominant inheritance pattern [McKusick, 1990: Mendelian inheritance in man]. A few isolated cases with autosomal recessive transmission have been described [Zlotogora et al., 1994: Am J Med Genet 49:261--262]. To our knowledge, no cases of X-linked colobomatous microphthalmia that are not a part of a syndrome or a multisystem disorder have been reported. In this study, we describe a genetic and clinical evaluation of a large pedigree in which colobomatous microphthalmia is segregating in an X-linked recessive fashion. Based on recombination breakpoint analysis, we have determined that the critical interval exists between markers DXS989 and DXS441, placing the disease locus on the proximal short arm or the proximal long arm of the X chromosome. Using linkage analysis, we obtained two-point lod scores of 2.71 at zero recombination with markers DXS1058, DXS6810, DXS1199, and DXS7132. Overlapping multipoint analysis established a broad maximum from marker DXS1068 to marker DXS7132, a region spanning approximately 28 cM. This study provides evidence for the presence of a new locus for colobomatous microphthalmia. PMID- 11391655 TI - FBN1 exon 2 splicing error in a patient with Marfan syndrome. AB - Mutations in FBN1 cause the autosomal dominant condition, Marfan syndrome. A single-base mutation that results in a skipping of exon 2 of FBN1 was found in a Marfan patient. By sequencing this proband's entire FBN1 gene and comparing the mutated DNA sequence with proband's unaffected family numbers, we confirmed this alteration was the causative mutation. The skipping of exon 2 creates a frameshift and premature termination codon, and forms a truncated fibrillin-1 composed only of 55 amino acids of N-terminus plus 45 nonsense amino acids. The mRNA transcription levels of the mutated FBN1 allele and the deposition of fibrillin-1 into extracellular matrix in fibroblast cells culture were assessed. PMID- 11391656 TI - New autosomal recessive cerebellar ataxia disorder in a large inbred Lebanese family. AB - A large inbred Lebanese pedigree with congenital spastic ataxia, microcephaly, optic atrophy, short stature, speech defect, abnormal osmiophilic pattern of skin vessels, cerebellar atrophy, and severe mental retardation transmitted as an autosomal recessive trait has been studied. None of the children had any evidence of a metabolic disease, and the analysis of respiratory chain complex abnormalities was unremarkable. Only one child had a history of perinatal difficulties. Differential diagnosis and the possibility that this disorder is a hitherto unreported one are discussed. PMID- 11391654 TI - Exclusion of linkage to the CDL1 gene region on chromosome 3q26.3 in some familial cases of Cornelia de Lange syndrome. AB - Cornelia de Lange Syndrome (CdLS) is a complex developmental disorder consisting of characteristic facial features, limb abnormalities, hirsutism, ophthalmologic involvement, gastroesophageal dysfunction, hearing loss, as well as growth and neurodevelopmental retardation. Most cases of CdLS appear to be sporadic. Familial cases are rare and indicate autosomal dominant inheritance. Several individuals with CdLS have been reported with chromosomal abnormalities, suggesting candidate genomic regions within which the causative gene(s) may lie. A CdLS gene location (CDL1) has been assigned to 3q26.3 based on phenotypic overlap with the duplication 3q syndrome (critical region 3q26.2-q27) and the report of a CdLS individual with a balanced de novo t(3;17)(q26.3;q23.1). It has been postulated that a gene within the dup3q critical region results in the CdLS when deleted or mutated. We have performed a linkage analysis to the minimal critical region for the dup3q syndrome (that encompasses the translocation breakpoint) on chromosome 3q in 10 rare familial cases of CdLS. Nineteen markers spanning a region of approximately 40 Mb (37 cM) were used. Results of a multipoint linkage analysis demonstrated total lod-scores that were negative across the chromosome 3q26-q27 region. In 4/10 families, lod-scores were less than -2 in the 2 cM region encompassing the translocation, while in the remaining 6/10 families, lod-scores could not exclude linkage to this region. These studies indicate that in some multicase families, the disease gene does not map to the CDL1 region at 3q26.3. PMID- 11391657 TI - Retinitis pigmentosa, growth hormone deficiency, and acromelic skeletal dysplasia in two brothers: possible familial RHYNS syndrome. AB - Here we report two brothers with retinitis pigmentosa, growth hormone deficiency, and acromelic skeletal dysplasia. We propose that their clinical picture is consistent with RHYNS syndrome (retinitis pigmentosa, hypopituitarism, nephronophthisis, and skeletal dysplasia) and that they represent the first instance of a familial occurrence of this syndrome. The presence of RHYNS in two siblings supports an autosomal recessive mode of inheritance; however, since all four known cases were male, an X-linked mode of inheritance cannot be excluded. The combination of clinical features found in these affected males is unique and supports the existence of RHYNS syndrome as a separate and distinct entity. PMID- 11391658 TI - Caucasian family with two independent mutations: 2594delC in BRCA1 and 5392delAG in BRCA2 gene. AB - Germline mutations in two breast cancer susceptibility genes, BRCA1 and BRCA2, predispose individuals to early onset breast and ovarian cancer. The frequency of mutations in these genes in the general population is very low. Therefore, the prior probability of finding any family with mutations in both genes is even lower. This study reports the presence of two mutations, one in BRCA1 and a second in BRCA2, in a single family with variable expression. The BRCA1 mutation, 2594delC, was identified first in the proband. Analysis on a related family member with early onset bilateral breast cancer for the same mutation was negative. Further analysis on the same individual led to the identification of a second germline mutation, 5392delAG in BRCA2 gene in this family. Without the knowledge of the second mutation in this family, many asymptomatic individuals would have been given a negative test result and be falsely reassured. Further analysis reveals differential expression of the two mutations. The spectrum of cancers as well as the age of onset is variable between the mutations and the generations. Finally, the study exemplifies the fact that molecular analysis of a genetically heterogeneous disease can be very complex and requires a team effort of the patients and their family members, genetic counselors or referring physicians as well as the personnel from the testing laboratory. PMID- 11391659 TI - Woman with UV hypersensitivity and a de novo unbalanced chromosome translocation. AB - We report a Japanese woman with de novo 6p monosomy and 10q trisomy [46,XX,der(6)t(6;10)(p25.1;q25.2)] whose clinical manifestations resemble those of xeroderma pigmentosum (XP) and Cockayne syndrome (CS), known as premature aging syndromes. She had a history of easy sunburning and presented a number of freckles and hypopigmented spots on her face as those of XP. Magnetic resonance imaging and computed tomography scanning demonstrated intracranial abnormalities like those seen in CS. DNA repair studies using the patient's fibroblasts demonstrated hypersensitive responses to ultraviolet (UV). XP, CS, and UV sensitive syndromes with photosensitivity disturbances have been known as DNA repair abnormalities. However, an association of 6p monosomy with these diseases has not been reported so far. Molecular analysis of the patient we described may contribute to the identification of novel DNA-repair-related gene(s) and/or to the senile mechanism. PMID- 11391660 TI - Klippel-Feil anomaly with Sprengel anomaly, omovertebral bone, thumb abnormalities, and flexion-crease changes: novel association or syndrome? AB - We report on a family with Klippel-Feil anomaly (KF), Sprengel anomaly, omovertebral bone, thumb abnormalities, and flexion-crease abnormalities. This combination of abnormalities does not fit into Holt-Oram syndrome, Wildervanck syndrome, oculo-auriculo-vertebral (Goldenhar) anomaly, or the VATER complex. Clinical aspects of a KF classification are discussed. The state of molecular research on KF is briefly reported. We conclude that this set of anomalies is a novel combination, probably representing pleiotropy of a single Mendelian gene. PMID- 11391661 TI - Human anotocephaly (aprosopus, acrania-synotia) in the Vilnius anatomical collection. AB - A genetic theory of "multifactorial" malformations, i.e., anomalies of blastogenesis or organogenesis, involving polygenic predisposition with morphogenetic threshold effect, was developed by Sewall Wright in the 1920s and remains an essential basis of birth defects biology. Because of the phenomenon of universality, i.e., the deployment of identical inductive, or pattern-forming, upstream molecular mechanisms during the earliest stages of mammalian morphogenesis, Wright's work on guinea pig otocephaly is highly pertinent to "corresponding," i.e., homologous malformations in humans. This concept is illustrated on the hand of a human fetus in the Vilnius (Lithuania) Pathological Museum with anotocephaly, i.e., anencephaly and otocephaly so severe as to correspond to Wright's guinea pig otocephaly grade 11 or 12. The observation also supports our apology for old museums and old books as repositories for anomalies, no less important for their rarity. PMID- 11391663 TI - Candidate region for cardio-facio-cutaneous syndrome. PMID- 11391664 TI - Separation of amniotic membranes after amniocentesis in an individual with the classic form of EDS and haploinsufficiency for COL5A1 expression. PMID- 11391665 TI - Mosaic (segmental) neurofibromatosis type 1 (NF1) and type 2 (NF2): no longer neurofibromatosis type 5 (NF5). PMID- 11391667 TI - Pulmonary artery sling in a case of trisomy 18. PMID- 11391666 TI - From DFNB2 to Usher syndrome: variable expressivity of the same disease. PMID- 11391669 TI - Chiral resolution. PMID- 11391670 TI - Direct resolution of (+/-)-ephedrine and atropine into their enantiomers by impregnated TLC. AB - Direct resolution of (+/-)-ephedrine and atropine into their enantiomers was achieved by normal-phase thin layer chromatography on silica gel plates impregnated with optically pure L-tartaric acid and L-histidine, respectively, as chiral selectors. The mobile phases enabling successful resolution were different combinations of acetonitrile-methanol-water. The spots were detected with iodine vapours and the detection limits were 2 and 6 microg, respectively, in terms of the racemate. The effects of concentration of the impregnating reagent, temperature and pH on resolution have been studied. PMID- 11391671 TI - Separation of amino acids, their derivatives and enantiomers by impregnated TLC. AB - The present state of TLC with respect to separation of amino acids, their different derivatives and their enantiomers by the technique of impregnation is discussed. The main approaches to impregnation viz. mixing of a suitable reagent with the adsorbent prior to plate-making, immersion of the untreated plate in the solution of impregnating reagent prior to development, and modification of the adsorbent, have been identified and discussed for each class of these compounds. The role of impregnation in resolving enantiomers or in improving the separation of mixtures of amino acids or their derivatives in terms of ion pairing, complex formation, ligand exchange or other steric interactions has been elaborated in each category. PMID- 11391672 TI - Determination of amino acid enantiomers in human urine and blood serum by gas chromatography-mass spectrometry. AB - Amino acid (AA) enantiomers were determined as N(O)-pentafluoropropionyl-(2) propyl esters by chiral gas chromatography-mass spectrometry (GC-MS) in 24 h samples of the urine of three healthy volunteers and in their blood sera. In urine the largest amounts were determined for D-Ser (64-199 micromol/day) and D Ala (24-138 micromol/day). In blood sera, D-Ala (2.3-4.2 micromol/L) and D-Ser (1.0-2.9 micromol/L) were most abundant. Varying amounts of the D-enantiomers of Thr, Pro, Asx, Glx, Phe, Tyr, Orn and Lys were also found, albeit not in all urines and sera. Further, enantiomers were quantified in urine samples of two volunteers fasting for 115 h. Quantities of renally excreted D-AAs decreased in fasting, although amounts of D-Ser (69 and 77 micromol/L urine) as well as other D-AAs were still detectable. Time-dependent analyses of urine showed that D-AAs are continuously excreted. PMID- 11391673 TI - Simultaneous microbatch screening of enantiorecognition on solid chiral selectors using selected mixtures of test-racemates: a case study on cellulose tris(alpha phenylpropionate) with configurational diversity. AB - An improved methodology for microbatch screening in the liquid-solid enantioselective adsorption of racemates by solid chiral selector is presented and illustrated by the evaluation of a series of six cellulose tris(alpha phenylpropionate) presenting a configurational diversity and cellulose tris (4 methylbenzoate). Analyses were performed on 5 mg scale of chiral selector and 150 microL of supernatant containing a mixture of three test-racemates. Fifteen test racemates were chosen according to their molecular diversity using a hierarchical clustering approach for seven selected three-dimensional molecular properties. The 15 racemates were sorted in five test mixtures composed of three racemates each according to the following constraints: each triplet of test-racemates can be analyzed without peak overlap on a commercially available chiral stationary phase (CSP). The designed five groups of three racemates may be used for the evaluation of other chiral selectors in view of the preparation of CSPs. This methodology improves the throughput of the evaluation and could be automated. PMID- 11391674 TI - Hybridation of different chiral separation techniques with ICP-MS detection for the separation and determination of selenomethionine enantiomers: chiral speciation of selenized yeast. AB - Enantioseparation and determination of selenomethionine enantiomers in selenized yeast was investigated using chiral separation techniques based on different principles, coupled on-line to inductively coupled plasma mass spectrometry (ICP MS) for selenium-specific detection. High performance liquid chromatography (HPLC) on a beta-cyclodestrin (beta-CD) column, cyclodextrin-modified micellar electrokinetic chromatography (CD-MEKC), gas chromatography (GC) on a Chirasil-L Val column, and HPLC on a Chirobiotic T column have been investigated as the chiral separation techniques. For HPLC separation on the beta-CD column, and also for CD-MEKC, selenomethionine enantiomers were derivatized with NDA/CN(-). For chiral separation by GC, selenomethionine enantiomers were converted into their N trifluoroacetyl (TFA)-O-alkyl esters. The developed hybridation methodologies are compared with respect to enantioselectivity, sensitivity and analysis time. The usefulness of the best-suited method [HPLC (Chirobiotic T)-ICP-MS] was demonstrated by its application to the successful chiral speciation of selenium and D-and L-selenomethionine content determination in selenized yeast. PMID- 11391675 TI - Simultaneous determination of D-lactic acid and 3-hydroxybutyric acid in rat plasma using a column-switching HPLC with fluorescent derivatization with 4-nitro 7-piperazino-2,1,3-benzoxadiazole (NBD-PZ). AB - A highly sensitive method for the determination of D-lactic acid and 3 hydroxybutyric acid (3-HB) in rat plasma was developed using high-performance liquid chromatography with octadecylsilica (ODS) connected to a chiral column. At first, (D + L)-lactic acid and 3-HB in the plasma were derivatized with a fluorescent reagent, 4-nitro-7-piperazino-2,1,3-benzoxadiazole (NBD-PZ), separated on the ODS column and determined fluorimetrically at 547 nm with 491 nm of excitation wavelength. During the separation step on the ODS, the peak fraction of (D + L)-lactate derivative was introduced directly to a phenylcarbamoylated beta-cyclodextrin chiral column by changing the flow of the eluent via a six-port valve. Then, D-lactate derivative was separated enantiomerically from the L-lactate derivative, and the enantiomeric ratio was determined from the chromatogram. Intra- and inter-day accuracy values for the determination of D-lactic acid in 10 microL of rat plasma were 97.8-109.2 and 98.4-109.9%, and those for 3-HB were 99.8-108.4 and 99.8-103.8%, respectively. The intra- and inter-day precision values were within 4.6 and 5.1% for D-lactic acid, and 2.7 and 2.4% for 3-HB, respectively. The detection limits for D-lactic acid and 3-HB were approximately 2.0 and 0.04 microM, respectively (signal-to noise ratio 3). The proposed method was applied to the plasma of diabetic rats induced by intraperitoneal administration of streptozotocin, and the significant increases of both D-lactic acid and 3-HB concentrations were observed in the diabetic rats as compared to the normal rats. PMID- 11391677 TI - Enantioseparation of aminoglutethimide and thalidomide by high performance liquid chromatography or supercritical fluid chromatography on mono-2 and mono-6-O pentenyl-beta-cyclodextrin-based chiral stationary phases. AB - Mono-2 and mono-6-O-pentenyl-beta-cyclodextrin (mono-2-pent-beta-CD and mono-6 pent-beta-CD), covalently linked to mercaptopropylsilica gel (thiol-Si) through thioether or sulfone linkage, reveal differentiated enantioselectivities in the separation of piperidine-2,6-dione-related drugs, namely aminoglutethimide and thalidomide, in supercritical fluid conditions. Supercritical fluid chromatographic resolution on completely defined mono-cyclodextrin derivative based chiral stationary phases (CSP) is a method of choice for the separation of aminoglutethimide but not effective for thalidomide. For both high performance liquid chromatography (HPLC) and supercritical fluid chromatography (SFC) conditions, the impact of the position, imposed to be 2 or 6 in our synthetic pathway, of the pentenyl moiety on one of the glucopyranosidics of the CD cage is of crucial importance in the chiral discrimination phenomenon. Additionally, the nature of the heteroatom present in the spacer arm between the CD and the silica gel, in this case thioether or sulfone functionality, is also essential for the chiral recognition mechanism(s) for the solute enantiomer. PMID- 11391676 TI - Reversed-phase planar chromatography of some enantiomeric amino acids and oxazolidinones. AB - The resolution of two enantiomeric amino acids and four structurally related oxazolidinones was evaluated by reversed-phase planar chromatography using both home-made microcrystalline cellulose triacetate (MCTA) layers and mobile phase additivies, such as beta-cyclodextrins (beta-Cy), modified alpha, beta, gamma cyclodextrins and bovine serum albumin (BSA), on commercially available RP 18W/F254 and SilC18-50/UV254 plates. All the enantiomers were partially or baseline resolved by at least one of the chiral phases tested. Densitograms of optical antipodes and their mixtures were measured on MCTA layers developed with hydroalcoholic solvents. PMID- 11391678 TI - Inspection of the reversal of enantiomer migration order in ligand exchange micellar electrokinetic capillary chromatography. AB - Enantiometers of D,L-phenylalanine were separated by capillary electrophoresis based on the principle of ligand exchange. Copper (II) complex of 4-hydroxy-L proline was used as chiral selector. The separation and the migration order of D- and L-phenylalanine were strongly affected by adding an anion surfactant sodium dodecylsulfate (SDS). Without SDS in the electrolyte, the separation was also carried out but the resolution was very small. With SDS added into the electrolyte, the resolution decreased with increasing concentration of SDS until 5.0 mM. When the concentration of SDS in the electrolyte was over 5.0 mM, inversion of the migration order of DL-phenylalanine was observed and the resolution was also increased with increasing concentration up to 20 mM. It was interesting to find that the inversion of the migration order took place not only in the enantioscparation but also in the positional isomers. A family of a fluorinated amino acid, o-, m- and p-fluoro-D,L-phenylalanine was separated and the inversion of the migration order is discussed. PMID- 11391679 TI - Enantioselective high-performance liquid chromatography of therapeutically relevant aminoalcohols as their fluorescent 1-naphthyl isocyanate derivatives. AB - A method for determining the enantiomers of 10 therapeutically relevant aminoalcohols using HPLC and precolumn derivatization was developed. Naphthyl isocyanate reacted with racemic aminoalcohols to form urea derivatives which were separated isocratically on a cellulose tris(3,5-dimethylphenylcarbamate) coated silica gel column, and detected fluorometrically in the lower ng mL(-1) range. The effluents can also be monitored at lower sensitivity, using an ultraviolet detector operated at 220 nm. PMID- 11391680 TI - Chiral resolution of flurbiprofen and ketoprofen enantiomers by HPLC on a glycopeptide-type column chiral stationary phase. AB - Vancomycin is an amphoteric, glycopeptide, macrocyclic antibiotic. When attached to 5 microspherical silica gel, vancomycin proved to be an effective chromatographic chiral stationary phase that could be used in the reversed-phase mode. In this study, a bonded vancomycin chiral stationary phase (Chirobiotic Vtrade mark) was investigated for the chiral liquid chromatography analysis of ketoprofen and flurbiprofen. The selectivity factor (alpha) and the chiral resolution factor (RS) of Chirobiotic Vtrade mark were evaluated first as a function of the buffer pH and molarity, and second as a function of organic modifier type and composition of the mobile phase. Four organic modifiers (tetrahydrofuran, 2-propanol, 1,4-dioxane and methanol) have been tested for their selectivity. Optimized conditions using 20% of tetrahydrofuran in ammonium nitrate (100 mM, pH 5) were selected for the enantioseparation of flurbiprofen and ketoprofen from their racemic forms. At pH 5, these acidic compounds are almost negatively charged, while the chiral selector possesses a positive charge allowing it to interact electrostatistically with the analytes. Using these chromatographic conditions, the column stability was excellent over several months of experiments. PMID- 11391681 TI - Concentration of enzymatically active prostate-specific antigen (PSA) in the extracellular fluid of primary human prostate cancers and human prostate cancer xenograft models. AB - BACKGROUND: Prostate-specific antigen (PSA) targeted prodrugs are under development in our laboratory. Concentrations of total PSA and enzymatically active PSA produced by various human prostate cancer xenograft models have not been well characterized. METHODS: The concentration of PSA secreted into the extracellular fluid (ECF) in normal human prostate tissue, primary prostate cancers obtained directly from patients, and serially passageable human prostate cancer xenografts (PC-82, LNCaP, LAPC-4) were determined using Tandem assays. Percent enzymatically active PSA in the ECF and in conditioned media was also determined using a previously validated assay employing a monoclonal antibody to the PSA catalytic site. In addition, the concentration and activity of PSA within sera from men with and without prostate cancer, as well as from tumor-bearing animals, was likewise assayed. RESULTS: Normal human prostate tissue and primary human prostate cancers have high concentrations of PSA in the ECF (i.e., 1600 2100 nM). The majority of this PSA is enzymatically active (i.e., 80-90%). Human PC-82 prostate cancer xenografts also have high concentrations of PSA in the ECF (624 +/- 360 nM), and the majority of this PSA is also enzymatically active (i.e., 66 +/- 4%). In contrast, much lower concentrations of PSA are found in the ECF from LNCaP (45 +/- 9 nM) and LAPC-4 (7.3 +/- 0.6 nM). Only a small portion of the total PSA isolated from DHT-containing, serum-free, conditioned media from these cell lines is enzymatically active (i.e., approximately 18%). While PSA was detected in all serum samples regardless of the type of host, no enzymatically active PSA was detected in any of these serum samples. CONCLUSIONS: Prostate cancers obtained directly from patients produce and secrete large amounts of PSA, the majority of which is highly enzymatically active. In contrast, while PSA was detected in the sera, none of this PSA was enzymatically active. This is also the case for the human PC-82 prostate cancer xenografts. In contrast, LNCaP and LAPC 4 human prostate cancer xenograft models secrete approximately 70-300-fold less PSA in the ECF than prostate cancers from patients and the majority of this PSA is enzymatically inactive. Also, the serum from these animals had detectable PSA, but none of this PSA was enzymatically active. Thus, these latter two prostate cancer models define the least and the PC-82, the most, optimized xenograft model for screening PSA targeted prodrugs. PMID- 11391682 TI - Neuroendocrine cells in human prostate over-express the anti-apoptosis protein survivin. AB - BACKGROUND: Neuroendocrine (NE) differentiation may be related to the growth and progression of prostate cancer, especially androgen-insensitive tumors. Recently the over-expression of a new anti-apoptosis protein, survivin, has attracted attention for its potential implication in many human cancers. The fact that NE cells in prostate are bcl-2 negative prompted us to investigate if the prostatic NE cells over-express survivin. METHODS: Double immunohistochemical staining and immunofluorescence of chromogranin A (CgA) and survivin were performed in 57 patients with localized prostate cancer who underwent radical prostatectomy. The terminal deoxynucleotidyl transferase (TDT)-mediated dUTP-digoxigenin nick end labeling (TUNEL) method was used for apoptosis detection in three prostate cancer specimens with NE differentiation. The relationship between NE differentiation and clinicopathological characteristics, disease progression as well as patient survival, were analyzed retrospectively. RESULTS: It was found that NE cells in both benign and malignant prostate tissues over-expressed the anti-apoptosis protein survivin. While apoptosis was detected in non-NE epithelial cells, all NE cells were negative for apoptosis detection. During the period of follow-up, 17 (63%) of 27 patients with NE differentiation had prostate cancer progression, while 12 (40%) of 30 patients without NE differentiation had systemic prostate cancer progression. 10 (37%) of 27 patients with NE differentiation died from prostate cancer during the period of follow up, while 6 (20%) of 30 patients without NE differentiation died from prostate cancer. However, none of these characteristics reached statistical significance, probably because of the small number of cases enrolled. CONCLUSIONS: This study discovers that all the prostatic NE cells express the new anti-apoptosis protein survivin. This provides a strong molecular basis for the hypothesis that NE cells may endure stressful conditions and escape from apoptosis. While our results suggest a trend of NE differentiation with poorer prognosis, the prognosis implication cannot be concluded due to our small sample size. PMID- 11391683 TI - Effects of Bowman-Birk inhibitor concentrate (BBIC) in patients with benign prostatic hyperplasia. AB - BACKGROUND: The Bowman-Birk inhibitor is a soybean-derived protease inhibitor that has anti-inflammatory and anticarcinogenic activities. METHODS: A Phase I trial of Bowman-Birk inhibitor concentrate (BBIC) in 19 male subjects with benign prostatic hyperplasia (BPH) and lower urinary tract symptoms (LUTS) has been performed. RESULTS: The results of the trial indicated that there was no dose limiting toxicity of BBIC. There was a statistically significant decrease in serum PSA levels in all BBIC-treated patients. Some BBIC-treated patients exhibited a relatively large reduction in serum PSA levels, ranging up to a 43% reduction. There was also a statistically significant decrease in serum triglyceride levels and a decrease in prostate volume in the treated patients. The scores recorded in response to a urinary symptom questionnaire indicated improved urinary activities in the BBIC-treated patients; however, the control subjects exhibited similar improvements in urinary activities during the course of the trial. CONCLUSIONS: The data obtained in this trial, particularly the data suggesting that BBIC treatment may lead to reduced serum PSA levels and reduced prostate volumes, suggest that a Phase II clinical trial of BBIC for the therapy of BPH is warranted. PMID- 11391684 TI - Why phase III trials of maximal androgen blockade versus castration in M1 prostate cancer rarely show statistically significant differences. AB - BACKGROUND: The meta-analysis of maximal androgen blockade (MAB) concluded that there is no survival advantage of MAB over castration alone. However, the results from the largest trials yield conflicting results. METHODS: The design and results of three trials were examined. RESULTS: Most studies were planned to detect an over-optimistic difference in survival and immature data were published. The survival curves show that statistical assumptions are not fulfilled. Excluding from the meta-analysis all trials where a negative impact of disease flare on survival could not be excluded resulted in no difference in survival between MAB and castration. CONCLUSIONS: Trials of MAB should be planned to detect differences of no more than 5-10% in median survival. The analyses should only be carried out on mature data and should take into account the possibility of a negative impact on survival due to disease flare if no anti androgen has been given initially with an LH-RH agonist. PMID- 11391685 TI - Endogenous nitric oxide-mediated relaxation and nitrinergic innervation in the rabbit prostate: the changes with aging. AB - BACKGROUND: Nitric oxide (NO) plays the key role in the non-adrenergic non cholinergic (NANC) nerve-mediated relaxation of prostate. We tried to determine whether nitrinergic innervation of the prostate is reduced with aging, and whether a reduction of this innervation alters the relaxant properties of prostatic tissue. METHODS: The prostate isolated from young rabbits at the age of 3 months and aged rabbits at the age of 24 months was used. Pharmacologic experiments using electrical field stimulation (EFS) were performed on strips of prostate. Nitrinergic nerves were identified histochemically by the presence of NADPH diaphorase reactivity. RESULTS: The prostate weighed 0.34 +/- 0.06 g in the young and 0.78 +/- 0.13 g in the aged (P < 0.01). Electrical stimulation caused frequency-dependent relaxation that was inhibited by N-nitro-L-arginine and increased by L-arginine. Maximum relaxant rates of noradrenaline-induced tone were 41.3 +/- 1.8% and 22.7 +/- 0.6% in the young and the aged (P < 0.01), respectively. Exogenous NO caused concentration-dependent relaxation of the prostate. Both relaxation induced by electrical stimulation and exogenous NO were inhibited by LY83583 (a guanylate cyclase inhibitor). In all specimens, NADPH diaphorase activity was observed in the smooth muscle layer of the prostatic stroma and capsule. However, NADPH diaphorase positive nerves were significantly less in the aged than in the young (P < 0.01). CONCLUSIONS: As a result of these findings, it is suggested that both NO-mediated relaxation and nitrinergic innervation are reduced with aging in this animal model. PMID- 11391686 TI - Anti-interleukin-6 monoclonal antibody induces regression of human prostate cancer xenografts in nude mice. AB - BACKGROUND: Despite clinical associations and in vitro data suggesting that autocrine interleukin-6 (IL-6) production contributes to prostate cancer progression or chemotherapy resistance, there have been no reports that explore the role of IL-6 on prostate tumors in vivo. In the present study, we investigated the effect of IL-6 inhibition on the growth of human prostate cancer xenografts in nude mice. METHODS: To determine if autocrine IL-6 production contributes to prostate cancer growth and chemotherapy resistance in vivo, xenografts of a human prostate cancer cell line that produces IL-6 (PC-3) were established in nude mice. The mice were randomly divided into four treatment groups: (1) saline (vehicle control) + murine IgG (isotype control); (2) etoposide + murine IgG; (3) saline + anti-IL-6 monoclonal antibody; and (4) etoposide + anti-IL-6 monoclonal antibody. Tumors were measured twice weekly during a 4-week treatment period. At the conclusion of the study, all mice were sacrificed, and in addition to final volume, tumors were evaluated for the degree of apoptosis by TUNEL analysis. RESULTS: Anti-IL-6 Ab (with saline or etoposide) induced tumor apoptosis and regression ( approximately 60% compared to initial tumor size). Etoposide alone did not induce tumor regression or apoptosis in this animal model, and there was no synergy between anti-IL-6 Ab and etoposide. CONCLUSIONS: These studies suggest that IL-6 contributes to prostate cancer growth in vivo, and that targeting IL-6 may contribute to prostate cancer therapy. PMID- 11391687 TI - Conference summary on prostate growth and aging, 13-15 September 2000. PMID- 11391688 TI - Incremental net benefit in randomized clinical trials. AB - There are three approaches to health economic evaluation for comparing two therapies. These are (i) cost minimization, in which one assumes or observes no difference in effectiveness, (ii) incremental cost-effectiveness, and (iii) incremental net benefit. The latter can be expressed either in units of effectiveness or costs. When analysing data from a clinical trial, expressing incremental net benefit in units of cost allows the investigator to examine all three approaches in a single graph, complete with the corresponding statistical inferences. Furthermore, if costs and effectiveness are not censored, this can be achieved using common two-sample statistical procedures. The above will be illustrated using two examples, one with censoring and one without. PMID- 11391689 TI - Optimal continuous sequential boundaries for monitoring toxicity in clinical trials: a restricted search algorithm. AB - Continuous monitoring of severe adverse experiences can ensure the timely termination of a clinical trial if the therapy is shown to be harmful. In this paper we present methods for choosing a stopping rule for continuous monitoring of toxicity in small trials. They are especially useful for small phase II trials of about 30 patients for monitoring a binary toxicity event that is observed relatively quickly compared to the efficacy outcome. In 1987 Goldman described an algorithm for computing the exact type I error rate (alpha) and power (1-beta) of a specified discrete stopping boundary for sequential monitoring of a study with a fixed maximum number of patients (N) to be enrolled on the experimental therapy. Only an upper boundary was used since trials are only terminated for an excess frequency of toxicity and not for a low rate. By repeated use of this algorithm a stopping rule can be identified which has nearly the chosen level of (alpha) and a reasonable power depending on the design parameters of the study. The work reported here embeds this earlier algorithm as a subroutine in a larger FORTRAN program which searches all boundaries that fulfil constraints on size and power, as specified by the user. The search is restricted so that only those boundaries with size in a small neighbourhood of the chosen alpha are examined and displayed if the power is above a set minimum. These restrictions reduce the number of boundaries examined to only 0.4 per cent of all possible boundaries, thus reducing running time to a practical few seconds. Many such boundaries exist, the one with the largest power can then be chosen for monitoring the trial. The average sample number (ASN) and the expected relative loss (ERL) are also computed. The criterion for choosing may also be based on small ASN or low ERL in addition to power and appropriate alpha. PMID- 11391690 TI - Long-term survivor mixture model with random effects: application to a multi centre clinical trial of carcinoma. AB - A mixture model incorporating long-term survivors has been adopted in the field of biostatistics where some individuals may never experience the failure event under study. The surviving fractions may be considered as cured. In most applications, the survival times are assumed to be independent. However, when the survival data are obtained from a multi-centre clinical trial, it is conceived that the environmental conditions and facilities shared within clinic affects the proportion cured as well as the failure risk for the uncured individuals. It necessitates a long-term survivor mixture model with random effects. In this paper, the long-term survivor mixture model is extended for the analysis of multivariate failure time data using the generalized linear mixed model (GLMM) approach. The proposed model is applied to analyse a numerical data set from a multi-centre clinical trial of carcinoma as an illustration. Some simulation experiments are performed to assess the applicability of the model based on the average biases of the estimates formed. PMID- 11391691 TI - Bias reduction for risk ratio and vaccine effect estimators. AB - We examine the structural bias for established estimators of vaccine effects on susceptibility and for newer estimates of vaccine effects on infectiousness. We then propose and analyse new bias corrections for vaccine effect estimators of both susceptibility and infectiousness, as well as their combined effect on infection transmission. Each estimator is evaluated empirically with computer simulations. Of the estimators examined in this paper, those with the least bias and root mean squared error are computed by adding one to the positive count in the placebo population. We also identify a source of bias for a standard Bayesian estimator of risk ratios. PMID- 11391692 TI - A general family of distributions for longitudinal dependence with special reference to event histories. AB - Event histories play an increasingly important role in medical studies. Examples include times between recurrences of tumours, as with bladder cancer, and between repeated infections, as with chronic granulotomous disease. A general method for generating new distributions is proposed by introducing an intensity function into a density. This procedure yields, as special cases, several distributions already proposed in the literature. The families of distributions based on the Pareto distribution are of particular interest for event history analysis because of their relationship to the Laplace transform of a gamma distribution. They can yield multivariate distributions, with longitudinal (serial) dependence by a procedure similar to updating in the Kalman filter and with uniform dependence in a similar way to copulas. For longitudinal dependence, several such updating procedures are proposed. PMID- 11391693 TI - Estimation of non-parametric multivariate risk functions in matched case-control studies with application to the assessment of interactions of risk factors in the study of cancer. AB - In epidemiological studies one is interested in investigating the probability of disease depending on risk factors and in particular in detecting interactions of risk factors. Within the setting of parametric logistic regression, interactions can be modelled only in a clumsy and limited way. Modelling the risk function non parametrically, estimating it, for example, by a smoothing (thin plate) spline is attractive as a more explorative approach. For prospective studies this amounts to smoothing within the framework and distributional assumptions of generalized regression models (for binary observations). Case-control studies as retrospective studies with exposure to risk factors being observed do not immediately fit into this setting. In the special case of one-to-one matched studies, however, there is an appropriate likelihood again within the range of generalized models. Inferences will be illustrated using simulated and real data. PMID- 11391694 TI - Mixed multivariate generalized linear models for assessing lower-limb arterial stenoses. AB - Experiments and observational studies often involve gathering information on several response variables, enabling us to model their dependence on observable predictor variables. Despite associations between the response variables, they are often analysed separately using general and generalized linear models. This paper investigates applications of multivariate regression analysis to improve the accuracy of predictions and decisions, in the specific context of diagnosing arterial stenoses in human legs. Two basic models are developed for this application, using (i) four binary responses and (ii) a mixture of two binary and two normal responses. The results clearly demonstrate the potential advantages offered by this approach. PMID- 11391695 TI - Bayesian model selection: analysis of a survival model with a surviving fraction. AB - We describe a methodology for model comparison in a Bayesian framework as applied to survival with a surviving fraction. This is illustrated using a case study of a randomized and controlled clinical trial investigating time until recurrence of depression. Posterior distributions are simulated using Metropolis-within-Gibbs Markov chain methods. Models reflecting the effects of covariates on the log odds of being in the surviving fraction, the log of the hazard rate, as well as both and neither are compared. Bayes factors for comparing the models are obtained by using the bridge sampling method of calculating normalizing constants. PMID- 11391696 TI - Bootstrap-type confidence intervals for quantiles of the survival distribution. AB - In this paper we outline and illustrate an easy to program method for analytically calculating both parametric and non-parametric bootstrap-type confidence intervals for quantiles of the survival distribution based on right censored data. This new approach allows for the incorporation of covariates within the framework of parametric models. The procedure is based upon the notion of fractional order statistics and is carried forth using a simple beta transformation of the estimated survival function (parametric or non-parametric). It is the only direct method currently available in the sense that all other methods are based on inverting test statistics or employing confidence intervals for other survival quantities. We illustrate that the new method has favourable coverage probabilities for median confidence intervals as compared to six other competing methods. PMID- 11391697 TI - Methods for testing equality of means of health care costs in a paired design study. AB - In this paper we propose five new tests for the equality of paired means of health care costs. The first two tests are the parametric tests, a Z-score test and a likelihood ratio test, both derived under the bivariate normality assumption for the log-transformed costs. The third test (Z-score with jack knife) is a semi-parametric Z-score method, which only requires marginal log normal assumptions. The last two tests are the non-parametric bootstrap tests: one is based on a t-test statistic, and the other is based on Johnson's modified t-test statistic. We conduct a simulation study to compare the performance of these tests, along with some commonly used tests when the sample size is small to moderate. The simulation results demonstrate that the commonly used paired t-test on the log-scale and the Wilcoxon signed rank for differences of the two original scales can yield type I error rates larger than the preset nominal levels. The commonly used paired t-test on the original data performs well with slightly skewed data, but can yield inaccurate results when two populations have different skewness. The likelihood ratio test, the parametric and semi-parametric Z-score tests all have very good type I error control with the likelihood ratio test being the best. However, the semi-parametric Z-score test requires less distributional assumptions than the two parametric tests. The percentile-t bootstrap test and bootstrapped Johnson's modified t-test have better type I error control than the paired t-test on the original-scale and Johnson's modified t-test, respectively. Combining with the propensity-score method, we can also apply the proposed methods to test the mean equality of two cost outcomes in the presence of confounders. Our two applications are from health services research. In the first one, we want to know the effect of Medicaid reimbursement policy change on outpatient health care costs. The second one is to evaluate the effect of a hospitalist programme on health care costs in an observational study, and the imbalanced covariates between intervention and control patients are taken into account using a propensity score approach. PMID- 11391698 TI - Neuronal and glial beta-secretase (BACE) protein expression in transgenic Tg2576 mice with amyloid plaque pathology. AB - We measured tissue distribution and expression pattern of the beta-site amyloid precursor protein (APP)-cleaving enzyme (BACE) in the brains of transgenic Tg2576 mice that show amyloid pathology. BACE protein was expressed at high levels in brain; at lower levels in heart and liver; and at very low levels in pancreas, kidney, and thymus and was almost absent in spleen and lung when assayed by Western blot analysis. We observed strictly neuronal expression of BACE protein in the brains of nontransgenic control mice, with the most robust immunocytochemical labeling present in the cerebral cortex, hippocampal formation, thalamus, and cholinergic basal forebrain nuclei. BACE protein levels did not differ significantly between control and transgenic mice or as a result of aging. However, in the aged, 17-month-old Tg2576 mice there was robust amyloid plaque formation, and BACE protein was also present in reactive astrocytes present near amyloid plaques, as shown by double immunofluorescent labeling and confocal laser scanning microscopy. The lack of astrocytic BACE immunoreactivity in young transgenic Tg2576 mice suggests that it is not the APP overexpression but rather the amyloid plaque formation that stimulates astrocytic BACE expression in Tg2576 mice. Our data also suggest that the neuronal overexpression of APP does not induce the overexpression of its metabolizing enzyme in neurons. Alternatively, the age-dependent accumulation of amyloid plaques in the Tg2576 mice does not require increased neuronal expression of BACE. Our data support the hypothesis that neurons are the primary source of beta-amyloid peptides in brain and that astrocytic beta-amyloid generation may contribute to amyloid plaque formation at later stages or under conditions when astrocytes are activated. PMID- 11391699 TI - Local synthesis of nuclear-encoded mitochondrial proteins in the presynaptic nerve terminal. AB - One of the central tenets in neuroscience has been that the protein constituents of distal compartments of the neuron (e.g., the axon and nerve terminal) are synthesized in the nerve cell body and are subsequently transported to their ultimate sites of function. In contrast to this postulate, we have established previously that a heterogeneous population of mRNAs and biologically active polyribosomes exist in the giant axon and presynaptic nerve terminals of the photoreceptor neurons in squid. We report that these mRNA populations contain mRNAs for nuclear-encoded mitochondrial proteins to include: cytochrome oxidase subunit 17, propionyl-CoA carboxylase (EC 6.4.1.3), dihydrolipoamide dehydrogenase (EC 1.8.1.4), and coenzyme Q subunit 7. The mRNA for heat shock protein 70, a chaperone protein known to be involved in the import of proteins into mitochondria, has also been identified. Electrophoretic gel analysis of newly synthesized proteins in the synaptosomal fraction isolated from the squid optic lobe revealed that the large presynaptic terminals of the photoreceptor neuron contain a cytoplasmic protein synthetic system. Importantly, a significant amount of the cycloheximide resistant proteins locally synthesized in the terminal becomes associated with mitochondria. PCR analysis of RNA from synaptosomal polysomes establishes that COX17 and CoQ7 mRNAs are being actively translated. Taken together, these findings indicate that proteins required for the maintenance of mitochondrial function are synthesized locally in the presynaptic nerve terminal, and call attention to the intimacy of the relationship between the terminal and its energy generating system. J. Neurosci. Res. 64:447-453, 2001. Published 2001 Wiley-Liss, Inc. PMID- 11391700 TI - Age- and concentration-dependent neuroprotection and toxicity by TNF in cortical neurons from beta-amyloid. AB - The induction of an inflammatory response and release of cytokines such as TNF may be involved in the age-related etiology of Alzheimer disease (AD). In the brain, microglia have been shown to produce a wide variety of immune mediators, including the pro-inflammatory cytokine tumor necrosis factor (TNF). We hypothesize that with age there is increased ability of microglia to produce TNF or that age decreases the neuroprotective effect of TNF against beta-amyloid (Abeta) toxicity in neurons. We investigated the effects of Abeta(1-40) on TNF secretion from forebrain cultures of microglia from embryonic, middle-age (9 month) and old (36-month) rats. Over the first 12 hr of exposure to 10 microM Abeta (1-40), microglia from embryonic and old rats increase TNF secretion, although microglia from middle-age rats did not produce detectable levels of TNF. When low concentrations of TNF are added to neurons together with Abeta (1-40) in the absence of exogenous antioxidants, neuroprotection for old neurons is significantly less than neuroprotection for middle-age neurons. In neurons from old rats, high levels of TNF together with Abeta are more toxic than in neurons from middle-age or embryonic rats. These results are discussed in relation to neuroprotection and toxicity of the age-related pathology of AD. PMID- 11391701 TI - Nongenomic antiapoptotic signal transduction by estrogen in cultured cortical neurons. AB - Estrogen replacement therapy in menopausal women has been suggested to be beneficial in preventing the progression of cognitive impairment in Alzheimer disease. We demonstrated previously that the phosphatidylinositol 3-kinase (PI3 K)/Akt signal transduction pathway plays a pivotal role on the neuroprotection provided by 17beta-estradiol against acute glutamate toxicity. In the present study, we investigated the mechanism of neuroprotection against apoptosis because acute glutamate toxicity predominantly induced necrosis. 17beta-estradiol provided neuroprotection against apoptosis induced by staurosporine. This neuroprotection was inhibited by pretreatment with a PI3-K inhibitor, LY294002. An estrogen receptor specific antagonist, ICI182780, also suppressed the neuroprotection provided by 17beta-estradiol. Western blotting analysis demonstrated that treatment with 17beta-estradiol induced the phosphorylation of Akt within 5 min, which was suppressed by pretreatment with LY294002 and ICI182780. Furthermore, 17beta-estradiol induced phosphorylation of the cAMP response element binding protein (CREB) at Ser(133) within 15 min and then upregulated Bcl-2 in a PI3-K/Akt-dependent manner. Because CREB is known to be a transcription factor for Bcl-2, these results suggest that 17beta-estradiol exerts its antiapoptotic effects by CREB phosphorylation and Bcl-2 upregulation via nongenomic activation of the PI3-K/Akt pathway in cultured cortical neurons. PMID- 11391702 TI - Alpha-motoneurons of the injured cervical spinal cord of the adult rat can reinnervate the biceps brachii muscle by regenerating axons through peripheral nerve bridges: combined ultrastructural and retrograde axonal tracing study. AB - Following our previous studies related to brachial plexus injury and repair, the present experimentation was designed to examine the ultrastructural features of those motoneurons of the locally injured cervical spinal cord of adult rats that were seen to regenerate into peripheral nerve (PN) bridges and to reinnervate nearby skeletal muscles. Here, the peripheral connection of the PN bridge was made with the biceps brachii (BB) muscle. Three months postsurgery, the spinal motoneurons labelled by retrograde axonal transport of horseradish peroxidase (HRP), after its injection into the BB, were selected on thick sections, using light microscopy, for the presence of dark amorphous granules of the HRP reaction product. Serial ultrathin sections were then made from the selected material. For the 10 labelled neurons studied, we examined the synaptic boutons present on the membrane of the neuronal soma. For five of them, we could observe three of the six types of synaptic boutons described for the alpha-motoneurons of the cat (S type with spherical vesicles, F-types with flattened vesicles, and C-type with subsynaptic cistern). The largest boutons (type C) are specific to alpha motoneurons. In comparison to normal material, we noticed a decrease in the number of boutons and an increase in the number of glial processes. After a transient phase of trophic changes, the reinnervated BB muscles showed a return of their fibers to nearly normal diameters as well as evidence of fiber type grouping. Simultaneous staining with silver and cholinesterase also revealed the presence of new motor endplates frequently contacted by several motoneurons. The present study indicates that, after a local spinal injury, typical alpha motoneurons can reinnervate a skeletal muscle by regenerating axons into the permissive microenvironment provided by a PN graft. These data offer prospects for clinical reconstruction of the brachial plexus after avulsion of one or several nerve roots. PMID- 11391703 TI - Neurofilament cytoskeleton disruption does not modify accumulation of trophic factor mRNA. AB - Previously we described a transgenic mouse model in which neurofilaments are sequestered in neuronal cell bodies and withheld from the axonal compartment. This model and other transgenic models with disrupted neurofilaments are used widely to investigate the role of the neurofilament cytoskeleton in normal neurons and in inherited or acquired diseases. To interpret such studies, it is important to establish whether the maldistribution of neurofilaments has major secondary consequences on the cell biology of the affected neurons. Notably, multiple perturbations of the nervous system simultaneously affect both the neuronal cytoskeleton and neurotrophin expression. To determine whether the expression of neurotrophic factors or their receptors is perturbed by a primary disruption in neurofilaments, we compared the accumulated mRNA levels for ciliary neuroptrophic factor (CNTF), nerve growth factor, neurotrophin 3, and the alpha CNTF receptor in mature transgenic mice and their littermate controls. Consistently with the prolonged survival of neurons expressing atypical or abnormally distributed neurofilaments, no obvious changes were observed for any of the mRNA species examined. PMID- 11391704 TI - Up-regulation of the peripheral-type benzodiazepine receptor expression and [(3)H]PK11195 binding in gerbil hippocampus after transient forebrain ischemia. AB - In mammalian CNS, the peripheral-type benzodiazepine receptor (PTBR) is localized on the outer mitochondrial membrane within the astrocytes and microglia. The main function of PTBR is to transport cholesterol across the mitochondrial membrane to the site of neurosteroid biosynthesis. The present study evaluated the changes in the PTBR density, gene expression and immunoreactivity in gerbil hippocampus as a function of reperfusion time after transient forebrain ischemia. Between 3 to 7 days of reperfusion, there was a significant increase in the maximal binding site density (B(max)) of the PTBR antagonist [(3)H]PK11195 (by 94-156%; P < 0.01) and PTBR mRNA levels (by 1.8- to 2.9-fold; P < 0.01). At 7 days of reperfusion, in the hippocampal CA1 (the brain region manifesting selective neuronal death), PTBR immunoreactivity increased significantly. Increased PTBR expression after transient forebrain ischemia may lead to increased neurosteroid biosynthesis, and thus may play a role in the ischemic pathophysiology. PMID- 11391705 TI - Nitric oxide synthase-positive neurons in the rat superior colliculus: colocalization of NOS with NMDAR1 glutamate receptor, GABA, and parvalbumin. AB - We analyzed the potential input and output components of nitric oxide synthase (NOS)-containing neurons in the rat superior colliculus (SC). To identify whether NOS-positive neurons receive glutamatergic input we investigated the colocalization of NOS with NMDA receptor subunit R1 (NMDAR1). In addition, to examine whether putative nitric oxide synthesizing neurons represent a neurochemically specific or distinct subpopulation of cells in the SC we studied the colocalization of NOS with the neurotransmitter GABA, the calcium-binding proteins parvalbumin, calbindin and calretinin and with neuropeptides such as somatostatin, substance P and neuropeptide Y. We found that 90% of NOS-positive neurons in the superficial layers of the rat SC express NMDAR1. Nearly 20% of the population of nitridergic neurons also expresses GABA and 15% of them express parvalbumin. NOS-positive neurons in the superior colliculus did not contain calretinin, calbindin or either of the neuropeptides tested. The results of this study show that the capacity for synthesizing NO in the SC is largely restricted to neurons that receive glutamatergic inputs and that some of these neurons express GABA or parvalbumin. PMID- 11391706 TI - In vitro model of microglial deramification: ramified microglia transform into amoeboid phagocytes following addition of brain cell membranes to microglia astrocyte cocultures. AB - Changes in the morphology of ramified microglia are a common feature in brain pathology and culminate in the appearance of small, rounded, microglia-derived phagocytes in the presence of neural debris. Here, we explored the effect of adding brain cell membranes on the morphology of alphaMbeta2-integrin (CD11b/CD18, CR3) positive microglia cultured on a confluent astrocyte substrate as an in vitro model of deramification. Addition of brain membranes led to a loss of microglial ramification, with full transformation to small, rounded, macrophages at 20-40 microg/ml. Time course studies showed a rapid response, with first effects at 1-3 hours, and full transformation at 24-48 hours. Removal of cell membranes and exchange of the culture medium led to a similarly rapid process of reramification. Comparison of cell membranes from different tissues at 20 microg/ml showed strong transforming effect for the brain, more moderate for kidney and liver, and very weak for spleen and skeletal muscle. Fluorescent labeling of brain membranes revealed uptake by almost all rounded macrophages, by a subpopulation of glial fibrillary acidic protein (GFAP)-positive astrocytes, but not by ramified microglia. Phagocytosis of inert fluorobeads did not lead to a transformation into macrophages but their phagocytosis was inhibited by brain membranes, pointing to a saturable uptake mechanism. In summary, addition of brain cell membranes and their phagocytosis leads to a rapid and reversible loss of ramification. The differences in transforming activity from different tissues and the absence of effect from phagocytosed fluorobeads suggest, however, the need for a second stimulus following the phagocytosis of cell debris. PMID- 11391707 TI - Activated microglia in the human glaucomatous optic nerve head. AB - To investigate the distribution and potential participation of microglia, the resident defense cells of the central nervous system, in the optic nerve head (ONH) in glaucoma, histological paraffin sections of optic nerves from normal and glaucoma patients with mild to advanced nerve damage were studied using double labeling immunohistofluorescence. A monoclonal antibody for HLA-DR, indicating activated microglia, was colocalized with antibodies for functional proteins. In normal ONHs, microglia do not contain TGF-beta2, COX-2, or TNF-alpha and are not positive for PCNA; however, in glaucomatous ONHs, microglia contain abundant TGF beta2, TNF-alpha, and PCNA. In glaucomatous eyes, a few microglia are usually positive for COX-2. In normal ONHs, there are rarely microglia containing TGF beta1, NOS-2, TSP, TIMP-2, and CD68, but, in glaucomatous tissue, a few microglia are positive from the prelaminar to the postlaminar regions. MMP-1, MMP-2, MMP-3, and MMP-14 are constitutively present in the perivascular microglia in normal ONHs and appear to be more abundant in glaucomatous tissue. COX-1, TNF-R1, TIMP 1, and c-fms are constitutively present in normal tissues and appear to be increased in microglia in the glaucomatous ONHs. HSP27 is not present in microglia. In glaucomatous ONHs, microglia become activated and phagocytic and produce cytokines, mediators, and enzymes that can alter the extracellular matrix. Our findings suggest that activated microglia may participate in stabilizing the tissue early in the disease process, but, as the severity of the glaucomatous damage increases, the activities of microglia may have detrimental consequences for the pathological course of glaucomatous optic neuropathy. PMID- 11391708 TI - Conduction of impulses by axons regenerated in a Schwann cell graft in the transected adult rat thoracic spinal cord. AB - Central nervous system axons regenerate into a Schwann cell implant placed in the transected thoracic spinal cord of an adult rat. The present study was designed to test whether these regenerated axons are capable of conducting action potentials. Following the transection and removal of a 4- to 5-mm segment of the thoracic spinal cord (T8-T9), a polymer guidance channel filled with a mixture of adult rat Schwann cells and Matrigel was grafted into a 4- to 5-mm-long gap in the transected thoracic spinal cord. The two cut ends of the spinal cord were eased into the guidance channel openings. Transected control animals received a channel containing Matrigel only. Three months after implantation, electrophysiological studies were performed. Tungsten microelectrodes were used for monopolar stimulation of regenerated axons within the Schwann cell graft. Glass microelectrodes were used to record responses in the spinal cord rostral to the stimulation site. Evoked responses to electrical stimulation of the axon cable were found in two out of nine Schwann cell-grafted animals. These responses had approximate latencies in the range of those of myelinated axons. No responses were seen in any of the Matrigel-grafted animals. Histological analysis revealed that the two cases that showed evoked potentials had the largest number of myelinated axons present in the cable. This study demonstrates that axons regenerating through Schwann cell grafts in the complete transected spinal cord can produce measurable evoked responses following electrical stimulation. PMID- 11391709 TI - Differential DM20 mRNA expression distinguishes two distinct patterns of spontaneous recovery from murine autoimmune encephalomyelitis. AB - Experimental autoimmune encephalomyelitis (EAE) is an animal model for multiple sclerosis (MS) mediated by T cells responding to CNS myelin proteins. Immunization of SWXJ mice with the immunodominant p139-151 peptide of myelin proteolipid protein (PLP) results in a relapsing-remitting pattern of EAE characterized by incomplete remyelination during clinical recovery. In the present study we observed two distinct clinical patterns of spontaneous remission during recovery from EAE, viz., sustained remission involving continuous neurologic improvement and aborted remission involving modest transient clinical improvement. We hypothesized that the ability to recover from autoimmune demyelination was directly linked to remyelination events that recapitulated developmental processes. Quantitative immunocytochemistry of CNS tissue showed decreased demyelination in mice undergoing sustained remission compared to mice undergoing aborted remission. Quantitative RT-PCR analysis showed elevated expression of DM20, the developmental isoform of PLP, in CNS tissue from mice undergoing sustained remission compared to mice undergoing aborted recovery. Moreover, DM20 expression was similarly elevated in CNS tissue from mice undergoing sustained recovery from EAE relapse. Our data indicate that expression of the developmental DM20 isoform of PLP is intimately associated with decreased demyelination and sustained clinical recovery from EAE. Thus, DM20 gene expression may provide an appropriate molecular target for promoting CNS remyelination and may serve as a useful marker for predicting clinical outcome and assessing the effectiveness of strategies aimed at promoting CNS tissue repair during autoimmune demyelinating disease. PMID- 11391710 TI - In vitro exposure to hydroxyurea reduces sickle red blood cell deformability. AB - Hydroxyurea is a drug that is used to treat some patients with sickle cell disease. We have measured the deformability of sickle erythrocytes incubated in hydroxyurea in vitro and found that hydroxyurea acts to decrease the deformability of these cells. The deformability of normal erythrocytes was not significantly affected by hydroxyurea except at very high concentrations. Hydroxyurea also did not consistently reduce the deformability of sickle erythrocyte ghosts. We propose that the decreased deformability, observed in vitro, is due to the formation of methemoglobin and other oxidative processes resulting from the reaction of hydroxyurea and oxyhemoglobin. Although the reaction with normal hemoglobin is similar to that of sickle hemoglobin, the sickle erythrocytes are affected more. We propose that the sickle erythrocyte membrane is more susceptible to the reaction products of the reaction of hemoglobin and hydroxyurea. An earlier report has shown that hydroxyurea increases the deformability of erythrocytes in patients on hydroxyurea. Taken together, these data suggest that the improved rheological properties of sickle erythrocytes in vivo are due to the elevated numbers of F cells [cells with fetal hemoglobin]. The presence of the nitrosyl hemoglobin or methemoglobin from the reaction with hydroxyurea may also benefit patients in vivo by reducing sickling. PMID- 11391711 TI - Extracellular accumulation of bioactive substances during preparation and storage of various platelet concentrates. AB - BACKGROUND: Side effects of platelet transfusion may be associated with infusion of bioactive substances. We therefore studied extracellular accumulation of histamine, plasminogen activator inhibitor (PAI)-1, vascular endothelial growth factor (VEGF), and interleukin (IL)-6 during preparation and storage of various platelet concentrates. METHODS: Twenty buffy-coat-derived platelet pools (BCPC) were prepared and stored in platelet additive solutions (PAS). Twelve apheresis platelet (APC) units were prepared using the COBE Spectra LRS, and 14 were prepared using the Fenwal Amicus Separator. After preparation half of the content was drawn from each APC unit. The normal ranges of the substances were determined in plasma from all donors, and the extracellular concentrations of the substances were determined in supernatants collected on days 0, 1, 3, 5, and 7 of storage from all platelet preparations. RESULTS: The platelet counts were not significantly different in BCPC units and APC units. The BCPC units had a significantly higher white cell count than the APC units (P < 0.0001), but the count was significantly higher in the Amicus APC units than in the COBE APC units (P < 0.0001). The extracellular histamine concentration was significantly (P < 0.001) increased in BCPC units after preparation and without further increase during storage, while there was no accumulation of histamine in APC units. After preparation the PAI-1 concentration was significantly (P < 0.02) higher in BCPC units than in APC units, but during storage PAI-1 increased significantly (P < 0.05) more in APC units than in BCPC units. Similarly, VEGF concentration was significantly (P < 0.05) higher in BCPC units than in APC units after preparation. During storage, however, VEGF increased more in BCPC units compared with COBE Spectra APC units (P < 0.05), but compared with Amicus Separator APC units only for the first 3 days of storage. At days 5 and 7 of storage the VEGF concentration was significantly higher in the Amicus APC units than in the COBE APC units (P < 0.05). IL-6 was not detectable in any of the concentrates after preparation or during storage. CONCLUSION: Platelet concentrates prepared by the apheresis method may contain less white cell derived bioactive substances than platelet concentrates prepared by the buffy-coat method. However, a substantial storage time dependent platelet derived bioactive substance accumulation takes place in all platelet concentrates tested, presumably due to platelet disintegration. PMID- 11391712 TI - Value of duplex and color doppler ultrasonography in the evaluation of orbital vascular flow and resistance in sickle cell disease. AB - The aim of the present study was to assess and to compare the orbital and retinal vascular flow dynamics and resistance in patients with homozygous sickle cell disease with controls by means of duplex and color Doppler ultrasonography. Forty six patients with homozygous sickle cell disease (SCD) and 20 healthy subjects were included in the study. None of the patients had objective signs of ocular involvement. Duplex and color Doppler ultrasonography of the ophthalmic, short posterior ciliary, and central retinal arterial flows of the both eyes were performed to assess peak systolic flow velocity (PSFV), end-diastolic flow velocity (EDFV), and mean flow velocity (MFV) through entire cardiac cycle with further calculation of resistive indices (RI) and pulsatility indices (PI). Ophthalmic arterial flow velocities were significantly increased in patients with SCD than in controls (P < 0.0001). Blood flow velocities of the central retinal artery were found to be significantly reduced (P < 0.0001) while RI and PI values were markedly higher (P < 0.02 and P < 0.03) in patients with SCD compared to controls. Reduction of retinal vascular flow velocities and increase of retinal vascular resistance were significantly related to the mean hemoglobin and hematocrit levels, red blood cell count, and mean corpuscular hemoglobin volume (P < 0.009, P < 0.01, P < 0.02, and P < 0.04, respectively). In conclusion, Doppler ultrasonography in patients with SCD who had no objective signs of ocular involvement allowed detection enhancement of ophthalmic flow velocities, reduced retinal flow velocities, and increased retinal vascular resistance, which are associated with haematological features. PMID- 11391713 TI - Hepatitis C virus infection and lymphoproliferative diseases: prospective study on 1,576 patients in France. AB - CONTEXT: Discordant data have been reported about the prevalence of hepatitis C virus (HCV) infection in patients with lymphoproliferative diseases and the putative role of HCV in lymphomagenesis. OBJECTIVE: To assess the prevalence of HCV infection in patients admitted to a hematology department in Paris, France. DESIGN: Prospective, controlled study. SETTING: University medical center. PATIENTS: 813 patients admitted to the Hematology department (164 B-cell non Hodgkin's lymphoma, 34 Hodgkin's diseases, 107 chronic lymphocytic leukemia, 54 multiple myeloma, 12 Waldenstrom's macroglobulinemia, 17 acute lymphoblastic leukemia, 6 hairy cell leukemia, 189 myeloproliferative diseases, 6 solid organ tumors, and 224 nonmalignant diseases) and 694 patients admitted to the Internal Medicine department (control group). MEASUREMENTS: All patients were tested for antibodies to HCV by a third-generation enzyme-linked immunosorbent assay. RESULTS: HCV antibodies were detected in 20 of 813 (2.46%) patients in Hematology including 11 of 394 (2.79%) patients with lymphoproliferative diseases, 3 of 164 (1.83%) B-cell non-Hodgkin's lymphoma, 2 of 107 (1.87%) chronic lymphocytic leukemia, 1 of 54 (1.85%) multiple myeloma, 1 of 189 (0.5%) myeloproliferative diseases, and 8 of 224 (3.57%) nonmalignant hematologic diseases. HCV antibodies were detected in 3 of 694 (0.43%) patients in the control group. HCV contamination preceded B-cell non-Hodgkin's lymphoma only in 2 of 3 HCV-positive patients. CONCLUSION: The prevalence of HCV infection was low (1.83%) in patients with B-cell non-Hodgkin lymphoma. HCV seems not to play a major role in the pathogenesis of B-cell lymphoma in France. Cofactors should be stressed to explain geographical discrepancies. PMID- 11391714 TI - Systemic anaplastic large-cell lymphoma: results from the non-Hodgkin's lymphoma classification project. AB - Anaplastic large-cell lymphoma (ALCL) is a heterogeneous process that may have a T-cell, B-cell, or indeterminant (null) phenotype and which may or may not express the anaplastic lymphoma kinase (ALK) oncoprotein. Because the clinical significance of these variants of ALCL is unclear, we evaluated the cases of ALCL T/null and ALCL-B identified in the Non-Hodgkin's Lymphoma Classification Project. We evaluated 1,378 cases of non-Hodgkin's lymphoma (NHL), and a consensus diagnosis of ALCL-T/null was made in 33 patients (2.4%) with a diagnostic accuracy of 85%. Compared to 96 patients with other forms of peripheral T-cell lymphoma (PTCL), those with ALCL-T/null were significantly younger, less likely to have advanced-stage disease or bone marrow involvement, more likely to have a low International Prognostic Index score, and had a significantly better survival. Among those with ALCL-T/null, there were no significant differences in the clinical features or survival on the basis of ALK expression. A consensus diagnosis of ALCL-B was made in 15 patients (1.1%), and the diagnostic accuracy was 67%. However, compared to 366 patients with other forms of diffuse large B-cell lymphoma (DLBCL), those with ALCL-B were no different with regard to clinical features or survival. We conclude that patients with ALCL-T/null have favorable prognostic features and excellent survival and should be separated from those with other forms of PTCL for prognostic and therapeutic purposes. In contrast, patients with ALCL-B appear to be similar to those with other forms of DLBCL. PMID- 11391715 TI - Molecular characteristics of pediatric patients with sickle cell anemia and stroke. AB - Cerebrovascular accidents (CVA) are serious complications of sickle cell anemia (SS) in children. Factors that predispose children to this complication are not well established. In an effort to elucidate the risk factors associated with CVA in SS, we have determined the alpha-globin genotype and the beta(S) haplotype of children with this complication. Among 700 children with SS followed at Children's Hospital of Michigan, 41 (6%) are on chronic transfusions because of stroke due to cerebral infarction. The mean age of patients with CVA at the time of stroke was 5.6 +/- 3.2 years (mean +/- SD). The male/female ratio was 2/3. Only 8 of 41 patients (19.5%) had one alpha-gene deletion, compared to the reported prevalence of 30% in African-Americans. None of the patients had two alpha-gene deletions, and two (5%) had five alpha-genes. These findings are different than those in our adult patients with SS, where the prevalence of alpha/-alpha and alphaalphaalpha/alphaalpha is 4% and <2%, respectively. Ten different beta(S)-haplotypes were detected in the patients studied. The majority of the patients (31%) were doubly heterozygous for the Ben/CAR haplotypes followed by Ben/Ben, Ben/Sen, and CAR/CAR haplotypes, respectively. The prevalence of these haplotypes, with the exception of the CAR/CAR haplotype, was higher in females than males. All the patients with CAR/CAR haplotype were males, had four alpha-genes, and ranked third in prevalence. Three patients were heterozygous for the Cameron haplotype. The Cameron and atypical haplotypes were more prevalent than reported in patients with SS at large. The data suggest that CVA in children seems to occur more frequently in females and in patients with certain beta(S) haplotype. alpha-Gene deletion seems to offer a protective effect against this complication. Neonates with four or more alpha-genes whose beta(S) haplotype is Ben/CAR, atypical, or CAR/CAR seem to be at a higher risk for CAV than other patients. A prospective study on a larger group of patients with or without CVA may clarify this issue. PMID- 11391716 TI - A common mutation in the gene for coagulation factor XIII-A (VAL34Leu): a risk factor for primary intracerebral hemorrhage is protective against atherothrombotic diseases. AB - The role of a common polymorphism in the factor XIII A-subunit gene (FXIII Val34Leu) has been recently investigated as a protective genetic factor against arterial and venous thrombosis. In addition, the less frequent Leu34 allele has been described as a risk factor for intracerebral hemorrhage. We evaluated the prevalence of this polymorphism by PCR in three case-control studies of patients diagnosed as having primary intracerebral hemorrhage (PCH, n = 130), coronary heart diseases (CHD, n = 240; myocardial infarction/no myocardial infarction, 120/120), and cerebrovascular diseases (CVD, n = 240; cerebral infarction/transient ischaemic attack, 120/120). The matched control groups consisted of patients admitted to the hospital without history of vascular disease. In addition, 200 healthy subjects were investigated. The frequency of the mutated allele (Leu34) was higher in patients with PCH than in controls (33.8% vs. 23.1%, P = 0.009) and lower in CHD and CVD patients compared to controls (18.1% vs. 25.2%, P = 0.010 and 17.3% vs. 24.2%, P = 0.011, respectively). Moreover, among the patients with CHD, the Leu34 allele was underrepresented in cases with myocardial infarction than without (12.9% vs. 23.3%, P = 0.004) and than in controls (12.9% vs. 25.2%, P < 0.001). Similar findings were obtained in patients with CVD comparing the cases with cerebral infarction versus cases with transient ischaemic attack (12.5% vs. 22.1%, P = 0.008) and versus controls (12.5% vs. 24.2%, P < 0.001). Finally, considering altogether the groups of ischaemic patients (CHD and CVD, n = 480), it was noted a trend towards a higher mean age of the clinical onset in homozygotes for the Leu allele than in the wild types (P = 0.078). This study indicates that in our population possession of the FXIII Val34Leu mutation predisposes to the occurrence of primary intracerebral hemorrhage and protects against cerebral and myocardial infarction. A wider modulatory role in the progression and onset of atherothrombotic diseases could be ascribed to FXIII Val34Leu. PMID- 11391717 TI - Molecular characterization of hemoglobin C in Thailand. AB - We describe hematologic and DNA characterization of 12 hemoglobin C heterozygotes and three compound heterozygotes for hemoglobin C and hemoglobin E found in Thailand. Amplification and DNA analysis of genomic DNA by the polymerase chain reaction procedure permitted the identification of the beta(C) mutation at codon 6 of beta-globin gene (beta 6; GAG-AAG). beta-Globin gene haplotype analysis demonstrated that all beta(C) globin genes detected in these Thai individuals were associated with the haplotype (+ - - - - - +), indicating a non-African origin of this abnormal hemoglobin in Thailand. On routine hemoglobin typing, hemoglobin C is usually mistakenly identified as hemoglobin E because of theirs similar mobilities on cellulose acetate electrophoresis. The simple DNA assay for hemoglobin C based on an allele-specific polymerase chain reaction for accurate diagnosis of hemoglobin C was therefore developed. PMID- 11391718 TI - An unusual presentation of extramedullary plasmacytoma occurring sequentially in the testis, subcutaneous tissue, and heart. AB - Extramedullary plasmacytoma (EMP) is a rare neoplasm of soft tissue that usually arises in the respiratory tract, nasal cavity, sinuses, and nasopharynx. It is even more uncommon for it to arise either in the testis or heart. We report the presentation of a case where plasmacytomas were found sequentially in the testis, subcutaneous tissue, and heart. EMP usually has a good prognosis except when it involves the heart. Our patient survived for only 15 months post autologous hematopoietic stem cell transplantation. PMID- 11391719 TI - Recurrent thromboembolic disease following splenectomy for pyruvate kinase deficiency. AB - We report a case of recurrent thromboembolic disease and chronic pulmonary hypertension in an adult patient with pyruvate kinase deficiency who underwent splenectomy as a child. Thromboembolism has been reported as a complication following splenectomy for various hereditary chronic hemolytic anemias. To our knowledge, this association has not been described in patients specifically with pyruvate kinase deficiency. Our patient presented at age 37 with recurrent pulmonary emboli, 36 years after splenectomy for severe hemolytic anemia. Work-up for other hypercoagulable states was negative. The mechanism for hypercoagulability in this condition is unclear but may involve a quantitative or qualitative change in disrupted thrombogenic red blood cell membranes that would normally be removed by the spleen. Clinicians should have a high index of suspicion for thrombotic events in these patients, as early diagnosis and treatment can reduce morbidity and mortality, and chronic anticoagulation may help prevent the sequelae of repeated thromboembolic events. PMID- 11391720 TI - Detection of human herpesvirus 6 and JC virus in progressive multifocal leukoencephalopathy complicating follicular lymphoma. AB - Progressive multifocal leukoencephalopathy (PML), a demyelinating infectious disease caused by JC virus (JCV), occurs almost exclusively in immunocompromised patients usually with malignant diseases. We report here a Japanese female with follicular lymphoma who subsequently developed PML. In addition to JCV, human herpesvirus 6 (HHV-6) was detected in the affected brain lesions of the patient by polymerase chain reaction and by in situ hybridization. HHV-6, recognized as a neurotropic virus, is known to be reactivated during immunosuppression and can cause fatal complications such as encephalitis/encephalopathy. It is likely that impaired immunity associated with lymphoma and the additional immunosuppression following cytopenia-inducing chemotherapies predisposed the patient to reactivated HHV-6 infection. Although it remains to be clarified whether HHV-6 plays an important role as a co-agent with JCV in causing demyelination of the brain, our observation alerts physicians to the possible association of HHV-6 with the pathogenesis of PML. PMID- 11391721 TI - Early onset Pneumocystis carinii pneumonia after allogeneic peripheral blood stem cell transplantation. AB - Pneumocystis carinii (P. carinii) is one of the major opportunistic pathogens responsible for hematopoietic stem cell transplantation (HSCT)-related pneumonias. Although trimethoprim-sulfamethoxazol (TMP/SMX) prophylaxis has been shown to prevent almost all P. carinii infections, 1%-2% of patients may still experience this complication. P. carinii pneumonia (PCP) is usually a late complication in patients receiving TMP/SMX prophylaxis, with most cases occurring later than 2 months post-transplant. We report a patient who developed early onset PCP after allogeneic peripheral blood stem cell transplantation (PBSCT) from an HLA-identical sibling donor. On day 12, the patient complained of dyspnea and cough. A chest X ray showed infiltrates in right upper lobe with bilateral pleural effusion. By the findings of Grocott stain on bronchoalveolar lavage fluid obtained on day 14, he was diagnosed as having PCP. Intravenous TMP/SMX failed to improve the lesion. This is the earliest onset PCP in the literature after HSCT despite the prophylactic administration of TMP/SMX before transplant. PMID- 11391723 TI - Adverse reactions related to rVIIa? PMID- 11391722 TI - Activated protein C and inflammation in human myocardium after heart surgery. AB - To assess possible interactions between inflammation and activation of the anticoagulant protein C system during post-ischemic reperfusion protein C, APC, neutrophil L-selectin expression and myocardial myeloperoxidase activity (MPO) were measured in 19 patients undergoing cardiopulmonary bypass. After reperfusion for 10 min, APC to protein C ratio (APC/PC) increased from pre-reperfusion value 1.43 +/- 0.12 (mean +/- SEM) to 2.25 +/- 0.29, p = 0.015. Negative correlations were observed between APC/PC and MPO activity (Spearman r -0.64, p = 0.007) and APC/PC and neutrophil L-selectin expression (r = -0.7, p = 0.007, demonstrating that post-ishemic protein C activation was associated with decreased neutrophil tissue sequestration. Thus, physiological protein C activation may be involved in regulation of the inflammatory injury during reperfusion of human ischemic coronary circulation. PMID- 11391725 TI - Erythroleukemia following erythropoietin therapy, extramedullary hematopoiesis, and splenectomy in a patient with myelofibrosis and myeloid metaplasia. PMID- 11391724 TI - Successful treatment with Rituximab for relapsing immune thrombocytopenic purpura (ITP) associated with low-grade non-Hodgkin's lymphoma. PMID- 11391726 TI - Iron absorption: relative importance of iron transport pathways. PMID- 11391727 TI - Development of acute erythremia from myelodysplastic syndrome after treatment with 1,25-dihydroxyvitamin D(3). PMID- 11391728 TI - Pulmonary embolism after pregnancy in a patient with polycythemia vera. PMID- 11391729 TI - Case of plasma-cell dyscrasia in a 13-year-old girl. PMID- 11391730 TI - Absence of HTLV-I infection in multi-transfused thalassemia patients in North India. PMID- 11391731 TI - A single mutation is responsible for the high prevalence of G6PD deficiency in the Vataliya Prajapatis--an endogamous caste group from Western India. PMID- 11391732 TI - Jumpers. PMID- 11391733 TI - End of lines and boxes. PMID- 11391734 TI - Open interconnected model of basal ganglia-thalamocortical circuitry and its relevance to the clinical syndrome of Huntington's disease. AB - The early stages of Huntington's disease (HD) present with motor, cognitive, and emotional symptoms. Correspondingly, current models implicate dysfunction of the motor, associative, and limbic basal ganglia-thalamocortical circuits. Available data, however, indicate that in the early stages of the disease, striatal damage is mainly restricted to the associative striatum. Based on an open interconnected model of basal ganglia-thalamocortical organization, we provide a detailed account of the mechanisms by which associative striatal pathology may lead to the complex pattern of motor, cognitive, and emotional symptoms of early HD. According to this account, the degeneration of a direct and several indirect pathways arising from the associative striatum leads to impaired functioning of: (1) the motor circuit, resulting in chorea and bradykinesia, (2) the associative circuit, resulting in abnormal eye movements, "frontal-like" cognitive deficits and "cognitive disinhibition," and (3) the limbic circuit, resulting in affective and psychiatric symptoms. When relevant, this analysis is aided by comparing the symptomatology of HD patients to that of patients with mild to moderate Parkinson's disease, since in the latter there is similar dysfunction of direct pathways but opposite dysfunction of indirect pathways. Finally, we suggest a potential novel treatment of HD and provide supportive evidence from a rat model of the disease. PMID- 11391736 TI - Plasticity of afferent fibers to striatal neurons bearing D1 dopamine receptors in Parkinson's disease. AB - The loss of dopaminergic neurons in the substantia nigra provokes a plasticity of corticostriatal synapses in Parkinson's disease (PD). The corticostriatal pathway nevertheless makes synapses with neurons bearing D1 dopamine receptors (D1R) and/or D2 dopamine receptors. At the ultrastructural level, we analyzed the morphological characteristics of synapses formed by afferent fibers making asymmetric contacts with the dendritic spines of neurons identified by D1R immunoreactivity, in the striatum of control subjects and PD patients. A quantitative analysis of the morphological characteristics of the synapses and of the number of perforated synapses (considered to be very active) was performed. In PD, a 50% increase in the number of perforated synapses making contact with D1R dendritic spines was observed, whereas no change in the number of perforated synapses on non-D1R spines was observed. The change in the number of perforated synapses on D1R dendrites was associated with a slight but nonsignificant increase in the surface area of the corticostriatal afferent fibers and the surface of the mitochondria in these fibers (+29.0% and +34.6%, respectively). This suggests a hyperactivity of corticostriatal fibers in contact with D1R bearing neurons of the direct pathway in the basal ganglia circuitry. Since stimulation of the direct pathway is thought to alleviate the clinical symptoms of PD, this suggests that the differences observed may be involved in compensatory mechanisms. PMID- 11391735 TI - Chronic L-DOPA administration is not toxic to the remaining dopaminergic nigrostriatal neurons, but instead may promote their functional recovery, in rats with partial 6-OHDA or FeCl(3) nigrostriatal lesions. AB - In this study, we have examined the effects of chronic L-3,4 dihydroxyphenylalanine (L-DOPA) administration on the remaining dopaminergic neurons in rats with 6-hydroxydopamine (6-OHDA) or buffered FeCl(3) partial lesions to the nigrostriatal tract. L-DOPA administration increased the turnover of dopamine in the striatum. L-DOPA administration for 1 week produced an increase in the level of striatal RTI-121 binding, a specific marker for dopamine uptake sites on the dopaminergic nerve terminals in the striatum. However, longer periods of L-DOPA treatment decreased the level of RTI-121 binding in the striatum. In the partial 6-OHDA lesion model, L-DOPA treatment had a time dependent effect on the number of neurons demonstrating a dopaminergic phenotype i.e., neurons that are tyrosine hyrdoxylase (TH)-immunopositive, on the lesioned side of the brain. In the first few weeks of treatment, L-DOPA decreased the number of TH-positive neurons but with long-term treatment, i.e., 24 weeks, L DOPA increased the number of neurons demonstrating a dopaminergic phenotype. Even in the buffered FeCl(3) infusion model, where the levels of iron were increased, L-DOPA treatment did not have any detrimental effects on the number of TH positive neurons on the lesioned side of the brain. Consequently, chronic L-DOPA treatment does not have any detrimental effects to the remaining dopaminergic neurons in rats with partial lesions to the nigrostriatal tract; indeed in the 6 OHDA lesion model, long-term L-DOPA may increase the number of neurons, demonstrating a dopaminergic phenotype. PMID- 11391737 TI - The tau A0 allele in Parkinson's disease. AB - Parkinson's disease (PD) is primarily an alpha-synucleinopathy, rather than a tauopathy, but there is evidence for an indirect association of tau with the pathogenetic process in PD. We therefore assessed the frequency in PD of the tau A0 allele, a dinucleotide repeat marker that has been associated with a sporadic tauopathy, progressive supranuclear palsy (PSP). We found the A0 allele to comprise 79.2% of 758 alleles from PD patients and 71.2% of 264 control alleles (P = 0.008). We also performed a meta-analysis of three previous reports, two of which failed to produce statistically significant results. Taken together, they also support a PD/A0 allelic association, even after correction for misdiagnosis of PSP as PD (P< 0.001). The A0/A0 genotype frequency in our patients (62.3%) did not differ significantly from that in controls (53.0%, P = 0.062), but the meta analysis, even after correction for misdiagnosis, showed a significant result, with P = 0.002. The frequency of A0 allele and the A0/A0 genotype were compatible with Hardy-Weinberg equilibrium. The frequency of the A0 allele and the A0/A0 genotype in our patients with familial PD was not significantly greater than in those with sporadic PD. We conclude that the tau protein may play a small role in the pathogenesis of PD and that biochemical characterization of this role may suggest opportunities for PD prophylaxis. PMID- 11391738 TI - Frequency of levodopa-related dyskinesias and motor fluctuations as estimated from the cumulative literature. AB - There is no clear consensus regarding the frequency (and hence, the risk), of dyskinesias or motor fluctuations during chronic levodopa therapy for Parkinson's disease (PD). Multiple clinical series have tabulated these frequencies since the advent of levodopa over 30 years ago. We were interested in determining: (1) the aggregate frequency figures in the existing literature; and (2) how clinical series from the early levodopa era, which included patients with longer durations of parkinsonism, compare to more recent (modern era) series. We searched MEDLINE for all English language publications reporting the cumulative frequency of levodopa-induced dyskinesias or motor fluctuations during discrete intervals of treatment. This generated 2,478 publications spanning 1966 through September 2000. Papers with appropriate titles or abstracts were reviewed; reference lists from published clinical series were a source of additional papers for review. This ultimately yielded 74 publications with adequate data, relating to 112 intervals of levodopa treatment. Series that included patients with PD-onset well before levodopa availability (pre-levodopa era) were separately analyzed from all subsequent series. Series were grouped by duration of levodopa therapy and the median frequencies of dyskinesias and motor fluctuations were tabulated for each group. The data were analyzed both with and without adjustment for the number (N) in each series. Among series containing pre-levodopa era patients, the median dyskinesia frequency was already 50% by 5-6 months of treatment. This contrasts with the modern era series where dyskinesias were reported later in treatment. The median dyskinesia frequency was slightly less than 40% by 4-6 years of levodopa therapy among modern era patients. Motor fluctuations (wearing-off) were not tabulated in most of the early levodopa series. Among modern era reports, motor fluctuations were nil during the first year of levodopa therapy but were experienced by approximately 40% of patients by 4-6 years of treatment. Similar results were found when the analyses were restricted to only prospective studies where levodopa motor complications were targeted outcome measures. The conclusions reached were: (1) patients from the pre-levodopa era experienced dyskinesias much earlier during levodopa treatment than modern era patients, perhaps because of longer durations of pre-existing PD; (2) in the present era, patients treated with levodopa therapy for 4-6 years have approximately a 40% likelihood of experiencing motor fluctuations and a risk of dyskinesias just short of 40%; and (3) these findings represent incident data and the prevalence of clinically important morbidity may be substantially less. PMID- 11391739 TI - Transdermal dopaminergic D(2) receptor agonist therapy in Parkinson's disease with N-0923 TDS: a double-blind, placebo-controlled study. AB - N-0923 is a non-ergot, dopaminergic D(2) agonist designed to be transdermally available. It has anti-parkinsonian effects when infused intravenously. An adhesive matrix patch was developed to deliver N-0923 transdermally (N-0923 TDS). In this phase II trial, we evaluated the effectiveness of various doses of N-0923 TDS at replacing levodopa. Eighty-five Parkinson's disease (PD) patients were randomized to placebo or one of four doses of N-0923 TDS for 21 days. Change in daily levodopa dose was the primary efficacy measure. Significantly greater reductions in levodopa dose were achieved as compared to placebo for the two highest doses of N-0923 TDS. Patients treated with 33.5 mg and 67 mg N-0923 TDS decreased levodopa use by 26% and 28%, vs. 7% for placebo. N-0923 TDS was safe and well tolerated. PMID- 11391740 TI - Long-term safety and efficacy of unilateral deep brain stimulation of the thalamus in essential tremor. AB - Our objective was to investigate the long-term safety and efficacy of unilateral deep brain stimulation (DBS) of the VIM nucleus of the thalamus in essential tremor. Forty-nine patients were evaluated for DBS between December 1993 and March 1998. Tremor was assessed by a clinical rating scale at 3 and 12 months and then yearly. Three patients were not implanted, seven were explanted prior to 24 months, 11 were lost to long-term follow-up, and three died from unrelated causes. Twenty-five patients were evaluated with follow-up greater than or equal to 2 years. The last postsurgical follow-up occurred on average 40.2 +/- 14.7 months after surgery. Tremor scores were significantly improved with stimulation on at the long-term follow-up as compared to baseline. There was no change in tremor scores from baseline to long-term follow-up with stimulation off. There was no significant change in any stimulus parameters from 3 months to the long term follow-up. Three patients had asymptomatic intracerebral hemorrhages and one patient had postoperative seizures. Stimulus-related adverse reactions were mild and easily controlled with changes in stimulus parameters. Device-related complications were common and required repeated surgical procedures. Unilateral DBS of the thalamus has long-term efficacy in some patients for treatment of essential tremor. However, this therapy is compromised by loss of efficacy in some patients and device complications which increase the risk of additional surgical procedures. PMID- 11391741 TI - Altered movement trajectories and force control during object transport in Huntington's disease. AB - Individuals with Huntington's Disease (HD) have difficulty grasping and transporting objects, however, the extent to which specific impairments affect their performance is unknown. The present study examined the kinematics and force coordination during transport of an object in 12 subjects with HD and 12 age matched controls. Subjects grasped an object between their thumb and index finger, transported it 25 cm forward, replaced and released it while their fingertip forces and the object's position were recorded. Five trials were performed with each of three weights (200 g, 400 g, and 800 g). While bradykinesia was evident in subjects with HD, this slowness was not consistently observed in all phases of the movement. The slowness of movement seen during the task appears to be due to impairments in sequencing and the movement strategies selected by the subjects. Compared to control subjects, subjects with HD produced highly curvilinear hand paths and more variable grip forces that were dependent on the weight of the object. Isometric force development and movement speed during transport were unaffected by the disease. The results suggest that prolonged task durations in subjects with HD are not necessarily due to slowness of movement, per se. These findings have clinical implications for understanding the task-specific nature of movement impairments in HD and developing effective intervention strategies. PMID- 11391742 TI - Quantitative assessment of daytime motor activity provides a responsive measure of functional decline in patients with Huntington's disease. AB - Voluntary motor impairment is a functionally important aspect of Huntington's disease (HD). Therefore, quantitative assessment of disturbed voluntary movement might be important in follow-up. We investigated the relation between quantitatively assessed daytime motor activity and symptom severity in HD and evaluated whether assessment of daytime motor activity is a responsive measure in the follow-up of patients. Sixty-four consecutive HD patients and 67 age- and sex matched healthy controls were studied. Daytime motor activity was recorded using a wrist-worn activity monitor that counts all movements during a period of five consecutive days. Patients were rated clinically for voluntary motor impairment, dyskinesias, posture & gait, depression, cognitive impairment and functional capacity. Follow-up was available from 40 patients (mean follow-up 2.0 years) and 29 controls (mean follow-up 5.9 years). Despite chorea, patients had less daytime motor activity than controls (P < 0.005). This hypokinesia correlated with impaired voluntary movements (r = 0.37; P < 0.01), disturbed posture & gait (r = 0.38; P < 0.005) and especially with reduced functional capacity (r = 0.51; P < 0.0005). During follow-up, hypokinesia remained unchanged in clinically stable patients, but became worse in those whose functional disability progressed (P < 0.005). Hypokinesia seems a core symptom of HD which is related to functional capacity. Actimetric assessment of hypokinesia is responsive to disease progression and can be used as an objective tool for follow-up. PMID- 11391743 TI - Bilateral internal globus pallidus stimulation for the treatment of spasmodic torticollis. AB - Three patients with spasmodic torticollis (ST) obtained substantial benefit from bilateral globus pallidus internus (GPi) stimulation. Progressive improvement in ST occurred over several months but residual cervical dystonia remained. These results corroborate those obtained by Krauss et al. on three patients with ST. PMID- 11391744 TI - Prospective clinical study of writer's cramp. AB - Sixty-five consecutive patients with writer's cramp were studied prospectively to evaluate clinical and demographic features, the number of writing hours per day at dystonia onset, and the existence of trigger events. Assessment of writing and drawing was done on a standardized test using categorical scales. The importance of subjective handicap and pain, of postural and action tremor, the abnormal features of hand grip and the occurrence of mirror dystonia (dystonia occurring in the dominant hand when writing with the other hand) were studied. Thirty-two patients had simple writer's cramp, whereas the others had complex writer's cramp with several activities other than writing involved. No significant differences were seen in age of onset, legibility, pain, and handicap in these two groups. Seven patients had a family history of focal dystonia and six of them had a complex form of writer's cramp. Mirror dystonia was seen in 29 patients and in some it appeared useful to distinguish dystonic movements from secondary compensatory strategies. PMID- 11391745 TI - Sudden daytime sleep onset in Parkinson's disease: polysomnographic recordings. AB - Sleep attacks in Parkinson's disease are controversially discussed. This paper describes a patient with Parkinson's disease suffering from sudden, irresistible onset of sleep during daytime. Medication included levodopa, entacapone, budipine, and cabergoline. Introduction of entacapone was the last therapeutic action preceding onset of sleep events, suggesting increased bioavailabilty of levodopa to be provocative in this case. In contrast to previous cases, the sudden sleep events were witnessed by clinical staff members and documented by polysomnographic and video recordings. Polysomnography during these sleep events remarkably showed abrupt slowing of EEG-background activity and occurrence of slow eye movements and K-complexes within 10 seconds after stable wakefulness. Within 60 seconds, the polysomnographic pattern proceeded to stable sleep stage 2. PMID- 11391746 TI - Comorbidity of the nonmotor symptoms of Parkinson's disease. AB - Many patients with Parkinson's disease (PD) have clinically significant anxiety, depression, fatigue, sleep disturbance, or sensory symptoms. The comorbidity of these nonmotor symptoms and their relationship to PD severity has not been extensively evaluated. Ninety- nine nondemented PD patients were evaluated with the following battery of tests: Beck Anxiety Inventory (BAI), Beck Depression Inventory (BDI), Fatigue Severity Scale (FSS), Pittsburgh Sleep Quality Index (PSQI), a sensory symptom questionnaire, Unified Parkinson's Disease Rating Scale (UPDRS), Hoehn & Yahr (H/Y) Stage, and the Schwab & England ADL scale (S/E). The comorbidity of the nonmotor symptoms and their relationship to PD severity was analyzed. Thirty-six percent of the study population had depression (BDI > or =10), 33% had anxiety (BAI > or =10), 40% had fatigue (FSS > 4), 47% had sleep disturbance (PSQI > 5), and 63% reported sensory symptoms. Only 12% of the sample had no nonmotor symptoms. Fifty-nine percent of the patients had two or more nonmotor symptoms, and nearly 25% had four or more. Increased comorbidity was associated with greater PD severity (P < 001). This study reveals that the nonmotor symptoms of PD frequently occur together in the same patients. Increased comorbidity of the five nonmotor symptoms was associated with greater PD severity. These results suggest that recognition of these diverse nonmotor symptoms may be enhanced by looking for others when one nonmotor symptom has been identified. PMID- 11391747 TI - Long-term mortality results of the randomized controlled study comparing bromocriptine to which levodopa was later added with levodopa alone in previously untreated patients with Parkinson's disease. AB - The present paper compares, in terms of mortality, two treatment regimens for Parkinson's disease (PD), i.e., bromocriptine later combined with levodopa versus levodopa only. Between 1982 and 1989, 60 PD patients (29 treated with levodopa alone [group D] and 31 receiving first bromocriptine followed by an association of bromocriptine + levodopa [group B/D]) were recruited. Data were updated in January 2000. Survival functions were estimated using Kaplan Meier product-limit method and comparison between the two groups with the log-rank test. Mortality was also compared with that of the general French population using standardized mortality ratios (SMRs). The mean duration of follow-up was 10.3 +/- 3.0 years. Seventeen patients died during the follow-up: nine in the group B/D and eight in the group D. The probability of survival at 10 years was 79.0% [95% confidence interval [CI]: 71.4-86.6] in group B/D and 72.9% [95% CI: 63.3-82.6] in group D. In comparison with the general French population, SMRs were not statistically different from 1, in the whole sample of PD patients (1.21, 95 % CI [0.71-1.95]), in group D (0.98 [0.42-1.93]), or in group B/D (1.53 [0.70-2.92]). In this population, we were unable to find any favourable effect of an early use of bromocriptine on mortality in PD in comparison with levodopa alone. PMID- 11391748 TI - Intravenous amantadine improves levadopa-induced dyskinesias: an acute double blind placebo-controlled study. AB - Experimental evidence suggests that glutamatergic receptor blockade may improve the motor response complications associated with long-term levodopa treatment in Parkinson's disease (PD) patients. Our objective was to evaluate the acute effect of amantadine, a noncompetitive antagonist of the N-methyl-D-aspartate (NMDA) receptor, on levodopa-induced dyskinesias, and to gain further insights into the antidyskinetic mechanism of this drug. Nine PD patients with motor fluctuations and severely disabling peak of dose dyskinesias received their first morning levodopa dose, followed by a 2-hour intravenous amantadine (200 mg) or placebo infusion, on two different days. Parkinsonian symptoms and dyskinesias were assessed every 15 minutes during the infusion and for 3 hours thereafter, while patients were taking their usual oral antiparkinsonian therapy, by means of Unified Parkinson's Disease Rating Scale (UPDRS, motor examination), tapping test, and a modified Abnormal Involuntary Movement Scale (AIMS). Intravenous amantadine acutely improved levodopa-induced dyskinesias by 50%without any loss of the anti-parkinsonian benefit from levodopa. This study confirms the antidyskinetic effect of amantadine and strengthens the rationale for using antiglutamatergic drugs in the treatment of parkinsonian motor fluctuations. PMID- 11391749 TI - Hemiballism with hyperglycemia and striatal T1-MRI hyperintensity: an autopsy report. AB - We report on an autopsy findings of a 92-year-old male with hemiballism hemichorea associated with hyperglycemia and striatal hyperintensity on T1 weighed magnetic resonance imaging (MRI), a recently described clinicoradiological syndrome. Histologically, the putamen contralateral to the hemiballism consisted of multiple foci of recent infarcts associated with reactive astrocytic and interneuronal response. Substrate responsible for the MRI signal changes is still inconclusive. PMID- 11391750 TI - Latah in Jakarta, Indonesia. AB - OBJECTIVE: To investigate the characteristics of latah in modern Indonesia; to determine whether contemporary latah resembles the syndrome described in the nineteenth century; to compare the syndrome of latah to other disorders featuring tics or exaggerated startle responses. BACKGROUND: Latah, described centuries ago in Malay people, is characterized by an exaggerated motor startle response, often with associated involuntary vocalizations, echolalia, echopraxia, and forced obedience. Modern latah has not been systematically studied. DESIGN AND METHODS: Persons with latah living in Jakarta, Indonesia, were investigated using a standardized, videotaped protocol. RESULTS: Fifteen women were studied. All had exaggerated startle to touch, and 10 to frightening words. Echolalia was seen in 10, echopraxia in 11, and forced obedience in 13. The startle response did not habituate, but instead worsened in response to repeated stimuli. Startle and associated symptoms were only partially suppressible in fewer than half. CONCLUSION: Modern latah resembles that described over a century ago. Latah resembles other disorders with exaggerated startle response, but is clinically distinct from Tourette's Syndrome. PMID- 11391751 TI - Jumping Frenchmen of Maine. PMID- 11391752 TI - The "Ragin' Cajuns" of Louisiana. PMID- 11391753 TI - Neurodegenerative Huntington-like disorder. PMID- 11391754 TI - Pontine MRI hyperintensities ("the cross sign") are not pathognomonic for multiple system atrophy (MSA). PMID- 11391755 TI - Seminal figures in the history of movement disorders: Sydenham, Parkinson, and Charcot: Part 6 of the MDS-sponsored history of Movement Disorders exhibit, Barcelona, June 2000. PMID- 11391756 TI - Parkinsonism-plus syndromes: Part 7 of the MDS-sponsored History of Movement Disorders exhibit, Barcelona, June 2000. PMID- 11391757 TI - History of Myoclonus: Part 8 of the MDS-sponsored History of Movement Disorders exhibit, Barcelona, June 2000. PMID- 11391758 TI - 69-year-old man with gait disturbance and Parkinsonism. PMID- 11391759 TI - Speech dysfluency exacerbated by levodopa in Parkinson's disease. AB - The role of dopamine in the modulation of speech fluency is complex. In this report we describe two patients with Parkinson's disease whose speech dysfluency was exacerbated by the administration of levodopa. In doing so, we extend the observation that dopaminergic mechanisms may be involved in the regulation of speech fluency. It is important for clinicians to recognize that, in some instances, dopaminergic replacement therapy may exacerbate an underlying dysfluency syndrome in PD. PMID- 11391760 TI - Parkinsonism after glycine-derivate exposure. AB - This 54-year-old man accidentally sprayed himself with the chemical agent glyphosate, a herbicide derived from the amino acid glycine. He developed disseminated skin lesions 6 hours after the accident. One month later, he developed a symmetrical parkinsonian syndrome. Two years after the initial exposure to glyphosate, magnetic resonance imaging revealed hyperintense signal in the globus pallidus and substantia nigra, bilaterally, on T2-weighted images. Levodopa/benserazide 500/125 mg daily provided satisfactory clinical outcome. PMID- 11391761 TI - Penicillamine-induced lethal status dystonicus in a patient with Wilson's disease. AB - A 37-year-old man with Wilson's disease is described, in whom the introduction of penicillamine therapy was followed after 3.5 weeks by the development of the status dystonicus with a fatal outcome. PMID- 11391762 TI - Hemichorea, moya-moya, and ulcerative colitis. AB - This is a case report linking chorea, colitis, and moya-moya. The clear involvement in the vasculopathy of the basal ganglia offers an obvious substrate for the movement disorder. PMID- 11391763 TI - Deep brain stimulation of the internal pallidum did not improve chorea in a patient with neuro-acanthocytosis. AB - We report the failure of bilateral globus pallidus internus deep brain stimulation to improve chorea in a patient with chorea-acanthocytosis. Prior to this surgery the patient had experienced a striking but short lived amelioration of symptoms with clozapine therapy. PMID- 11391764 TI - Permanent cerebellar toxicity of cytosine arabinoside (Ara C) in a young woman. AB - This report provides the first videotape example of a patient with permanent cerebellar ataxia from high dose cytosine arabinoside. This case was unusual in that the patient was young, the effects are seemingly permanent and severe, and corticospinal tract damage also occurred. Early recognition to stop drug administration remains the only method to reduce risk. PMID- 11391765 TI - Restless legs syndrome in an Asian population: A study in Singapore. AB - In face-to-face interviews, we examined 157 consecutive individuals aged 55 years and older, selected from the general population in Singapore, and 1,000 consecutive individuals aged 21 years and older, from a primary healthcare center. Based on the IRLSSG criteria, the prevalence of restless leg syndrome (RLS) was 0.6% and 0.1%, respectively. PMID- 11391766 TI - Piribedil for restless legs syndrome: a pilot study. AB - Thirteen consecutive patients with restless legs syndrome (RLS) were treated with piribedil and were rated using an RLS rating scale (0-10) and subjective response (0-100%); 11/13 (85%) had improvement of their mean RLS scores with subjective response ranging from 30% to 100% (mean 74.6%). This pilot study suggests that piribedil is effective for RLS. PMID- 11391767 TI - Tuberculous meningitis and dystonia. PMID- 11391769 TI - Proline-induced hinges in transmembrane helices: possible roles in ion channel gating. AB - A number of ion channels contain transmembrane (TM) alpha-helices that contain proline-induced molecular hinges. These TM helices include the channel-forming peptide alamethicin (Alm), the S6 helix from voltage-gated potassium (Kv) channels, and the D5 helix from voltage-gated chloride (CLC) channels. For both Alm and KvS6, experimental data implicate hinge-bending motions of the helix in an aspect of channel gating. We have compared the hinge-bending motions of these TM helices in bilayer-like environments by multi-nanosecond MD simulations in an attempt to describe motions of these helices that may underlie possible modes of channel gating. Alm is an alpha-helical channel-forming peptide, which contains a central kink associated with a Gly-x-x-Pro motif in its sequence. Simulations of Alm in a TM orientation for 10 ns in an octane slab indicate that the Gly-x-x-Pro motif acts as a molecular hinge. The S6 helix from Shaker Kv channels contains a Pro-Val-Pro motif. Modeling studies and recent experimental data suggest that the KvS6 helix may be kinked in the vicinity of this motif. Simulations (10 ns) of an isolated KvS6 helix in an octane slab and in a POPC bilayer reveal hinge-bending motions. A pattern-matching approach was used to search for possible hinge bending motifs in the TM helices of other ion channel proteins. This uncovered a conserved Gly-x-Pro motif in TM helix D5 of CLC channels. MD simulations of a model of hCLC1-D5 spanning an octane slab suggest that this channel also contains a TM helix that undergoes hinge-bending motion. In conclusion, our simulations suggest a model in which hinge-bending motions of TM helices may play a functional role in the gating mechanisms of several different families of ion channels. PMID- 11391770 TI - Allosteric free energy changes at the alpha 1 beta 2 interface of human hemoglobin probed by proton exchange of Trp beta 37. AB - The energetic changes that occur on ligand binding in human hemoglobin have been investigated by measurements of the exchange rates of the indole proton of Trpbeta37(C3). The Trpbeta37 residues are located in helices C of the beta subunits and are involved in contacts with the segments FG of the alpha-subunits at the interdimeric alpha1beta2 and alpha2beta1 interfaces of the hemoglobin tetramer. In the quaternary structure change that accompanies ligand binding to hemoglobin, these contacts undergo minimal changes in relative orientation and in packing, thereby acting as hinges, or flexible joints. The exchange rates of the indole proton of Trpbeta37(C3) were measured by nuclear magnetic resonance spectroscopy, in both deoxygenated and ligated hemoglobin. The results indicate that, at 15 degrees C, the exchange rate is increased from 9.0. 10(-6) to 3.3. 10(-4) s(-1) upon ligand binding to hemoglobin. This change suggests that the structural units at the hinge regions of the alpha1beta2/alpha2beta1 interfaces containing Trpbeta37(C3) are specifically stabilized in unligated hemoglobin, and experience a change in structural free energy of approximately 4 kcal/(mol tetramer) upon ligand binding. Therefore, the hinge regions of the alpha1beta2/alpha2beta1 interfaces could play a role in the transmission of free energy through the hemoglobin molecule during its allosteric transition. PMID- 11391771 TI - How to guarantee optimal stability for most representative structures in the Protein Data Bank. AB - We proposed recently an optimization method to derive energy parameters for simplified models of protein folding. The method is based on the maximization of the thermodynamic average of the overlap between protein native structures and a Boltzmann ensemble of alternative structures. Such a condition enforces protein models whose ground states are most similar to the corresponding native states. We present here an extensive testing of the method for a simple residue-residue contact energy function and for alternative structures generated by threading. The optimized energy function guarantees high stability and a well-correlated energy landscape to most representative structures in the PDB database. Failures in the recognition of the native structure can be attributed to the neglect of interactions between different chains in oligomeric proteins or with cofactors. When these are taken into account, only very few X-ray structures are not recognized. Most of them are short inhibitors or fragments and one is a structure that presents serious inconsistencies. Finally, we discuss the reasons that make NMR structures more difficult to recognizeCopyright 2001 Wiley-Liss, Inc. PMID- 11391772 TI - Structure of the adenovirus E4 Orf6 protein predicted by fold recognition and comparative protein modeling. AB - To facilitate investigation of the molecular and biochemical functions of the adenovirus E4 Orf6 protein, we sought to derive three-dimensional structural information using computational methods, particularly threading and comparative protein modeling. The amino acid sequence of the protein was used for secondary structure and hidden Markov model (HMM) analyses, and for fold recognition by the ProCeryon program. Six alternative models were generated from the top-scoring folds identified by threading. These models were examined by 3D-1D analysis and evaluated in the light of available experimental evidence. The final model of the E4 protein derived from these and additional threading calculations was a chimera, with the tertiary structure of its C-terminal 226 residues derived from a TIM barrel template and a mainly alpha-nonbundle topology for its poorly conserved N-terminal 68 residues. To assess the accuracy of this model, additional threading calculations were performed with E4 Orf6 sequences altered as in previous experimental studies. The proposed structural model is consistent with the reported secondary structure of a functionally important C-terminal sequence and can account for the properties of proteins carrying alterations in functionally important sequences or of those that disrupt an unusual zinc coordination motif. PMID- 11391773 TI - Cryptic epitopes in N-terminally truncated prion protein are exposed in the full length molecule: dependence of conformation on pH. AB - Prion diseases are diseases of protein conformation. Structure-dependent antibodies have been sought to probe conformations of the prion protein (PrP) resulting from environmental changes, such as differences in pH. Despite the absence of such antibodies for full-length PrP, a recombinant Fab (D13) and a Fab derived from mAb 3F4 showed pH-dependent reactivity toward epitopes within the N terminus of N-terminally truncated PrP(90-231). Refolding and maintaining this protein at pH > or =5.2 before immobilization on an ELISA plate inhibited reactivity relative to protein exposed to pH < or =4.7. The reactivity was not affected by pH changes after immobilization, showing retention of conformation after binding to the plate surface, although guanidine hydrochloride at 1.5-2 M was able to expose the cryptic epitopes after immobilization at pH > or =5.2. The alpha-helical CD spectrum of PrP(90-231) refolded at pH 5.5 was reduced somewhat by these pH changes, with a minor shift toward beta-sheet at pH 4 and then toward coil at pH 2. No covalent changes were caused by the pH differences. This pH dependence suggests titration of an acidic region that might inhibit the N terminal epitopes. A similar pH dependence for a monoclonal antibody reactive to the central region identified an acidic region incorporating Glu152 as a significant participant. PMID- 11391774 TI - Silk fibroin: structural implications of a remarkable amino acid sequence. AB - The amino acid sequence of the heavy chain of Bombyx mori silk fibroin was derived from the gene sequence. The 5,263-residue (391-kDa) polypeptide chain comprises 12 low-complexity "crystalline" domains made up of Gly-X repeats and covering 94% of the sequence; X is Ala in 65%, Ser in 23%, and Tyr in 9% of the repeats. The remainder includes a nonrepetitive 151-residue header sequence, 11 nearly identical copies of a 43-residue spacer sequence, and a 58-residue C terminal sequence. The header sequence is homologous to the N-terminal sequence of other fibroins with a completely different crystalline region. In Bombyx mori, each crystalline domain is made up of subdomains of approximately 70 residues, which in most cases begin with repeats of the GAGAGS hexapeptide and terminate with the GAAS tetrapeptide. Within the subdomains, the Gly-X alternance is strict, which strongly supports the classic Pauling-Corey model, in which beta sheets pack on each other in alternating layers of Gly/Gly and X/X contacts. When fitting the actual sequence to that model, we propose that each subdomain forms a beta-strand and each crystalline domain a two-layered beta-sandwich, and we suggest that the beta-sheets may be parallel, rather than antiparallel, as has been assumed up to now. PMID- 11391775 TI - Cooperative helix stabilization by complex Arg-Glu salt bridges. AB - Among the interactions that stabilize the native state of proteins, the role of electrostatic interactions has been difficult to quantify precisely. Surface salt bridges or ion pairs between acidic and basic side chains have only a modest stabilizing effect on the stability of helical peptides or proteins: estimates are roughly 0.5 kcal/mol or less. On the other hand, theoretical arguments and the occurrence of salt bridge networks in thermophilic proteins suggest that multiple salt bridges may exert a stronger stabilizing effect. We show here that triads of charged side chains, Arg(+)-Glu(-)-Arg(+) spaced at i,i+4 or i,i+3 intervals in a helical peptide stabilize alpha helix by more than the additive contribution of two single salt bridges. The free energy of the triad is more than 1 kcal/mol in excess of the sum of the individual pairs, measured in low salt concentration (10 mM). The effect of spacing the three groups is severe; placing the charges at i,i+4 or i,i+3 sites has a strong effect on stability relative to single bridges; other combinations are weaker. A conservative calculation suggests that interactions of this kind between salt bridges can account for much of the stabilization of certain thermophilic proteins. PMID- 11391776 TI - Generalized comparative modeling (GENECOMP): a combination of sequence comparison, threading, and lattice modeling for protein structure prediction and refinement. AB - An improved generalized comparative modeling method, GENECOMP, for the refinement of threading models is developed and validated on the Fischer database of 68 probe-template pairs, a standard benchmark used to evaluate threading approaches. The basic idea is to perform ab initio folding using a lattice protein model, SICHO, near the template provided by the new threading algorithm PROSPECTOR. PROSPECTOR also provides predicted contacts and secondary structure for the template-aligned regions, and possibly for the unaligned regions by garnering additional information from other top-scoring threaded structures. Since the lowest-energy structure generated by the simulations is not necessarily the best structure, we employed two structure-selection protocols: distance geometry and clustering. In general, clustering is found to generate somewhat better quality structures in 38 of 68 cases. When applied to the Fischer database, the protocol does no harm and in a significant number of cases improves upon the initial threading model, sometimes dramatically. The procedure is readily automated and can be implemented on a genomic scale. PMID- 11391777 TI - Protein flexibility predictions using graph theory. AB - Techniques from graph theory are applied to analyze the bond networks in proteins and identify the flexible and rigid regions. The bond network consists of distance constraints defined by the covalent and hydrogen bonds and salt bridges in the protein, identified by geometric and energetic criteria. We use an algorithm that counts the degrees of freedom within this constraint network and that identifies all the rigid and flexible substructures in the protein, including overconstrained regions (with more crosslinking bonds than are needed to rigidify the region) and underconstrained or flexible regions, in which dihedral bond rotations can occur. The number of extra constraints or remaining degrees of bond-rotational freedom within a substructure quantifies its relative rigidity/flexibility and provides a flexibility index for each bond in the structure. This novel computational procedure, first used in the analysis of glassy materials, is approximately a million times faster than molecular dynamics simulations and captures the essential conformational flexibility of the protein main and side-chains from analysis of a single, static three-dimensional structure. This approach is demonstrated by comparison with experimental measures of flexibility for three proteins in which hinge and loop motion are essential for biological function: HIV protease, adenylate kinase, and dihydrofolate reductase. PMID- 11391778 TI - Differential behavioral responses to cocaine are associated with dynamics of mesolimbic dopamine proteins in Lewis and Fischer 344 rats. AB - Differential behavioral and biochemical responses to drugs of abuse may reflect genetic makeup as suggested by studies of inbred Lewis (LEW) and Fischer 344 (F344) rats. We investigated locomotor activity, stereotypy signs, and levels of specific proteins in the nucleus accumbens (NAc) and ventral tegmental area (VTA) in these strains at baseline and following chronic administration of cocaine (30 mg/kg/day for 14 days). Using Western blot analysis, we replicated our previous findings of baseline strain differences and found lower levels of DeltaFosB immunoreactivity in NAc of F344 vs. LEW rats. F344 rats showed greater baseline locomotor activity, sniffing, and grooming compared to LEW rats. Chronic cocaine increased DeltaFosB levels in NAc in both strains, whereas adaptations in other proteins were induced in F344 rats only. These included reduced levels of tyrosine hydroxylase (TH) in NAc and increased TH and glial fibrillary acidic protein (GFAP) immunoreactivity in VTA. Chronic cocaine led to greater increases in overall stereotypy in F344 vs. LEW rats and decreased exploratory behaviors in LEW rats. Opposing effects by strain were seen in locomotor activity. Whereas F344 rats showed higher initial activity levels that decreased with cocaine exposure (tolerance), LEW rats showed increased activity over days (sensitization) with no strain differences seen at 14 days. Further, conditioned locomotor activation to vehicle injections was greater in F344 vs. LEW rats. These results suggest that behavioral responsiveness to chronic cocaine exposure may reflect dynamics of mesolimbic dopamine protein levels and demonstrate the role of genetic background in responsiveness to cocaine. PMID- 11391779 TI - Defective hippocampal mossy fiber long-term potentiation in endothelial nitric oxide synthase knockout mice. AB - Mossy fiber long-term potentiation (mfLTP) was compared in hippocampal slices prepared from wild-type mice and mice lacking functional endothelial nitric oxide synthase (eNOS(-/-) mice) using field potential recording. In the presence of D-2 amino-5-phosphonovaleric acid (AP5, 50 microM), the mfLTP induced by tetanic stimulation (100 Hz, 1 sec) was significantly reduced in knockouts (n = 8) in comparison with wild-type (n = 8). Similarly, potentiation induced by forskolin (30 microM) or 8-bromo-cyclic adenosine monophosphate (8-Br-cAMP, 100 microM) was less pronounced in knockouts. However, in wild-types the mfLTP-induced in the presence of the nonselective pharmacological inhibitor of NOS (N-nitro-L Arginine, 100 microM, n = 6) was not significantly different from control (n = 8). Thus, eNOS is not directly involved in mfLTP, but lack of eNOS during development leads to a deficit downstream of adenylyl cyclase. PMID- 11391780 TI - Changes in cannabinoid CB(1) receptors in striatal and cortical regions of rats with experimental allergic encephalomyelitis, an animal model of multiple sclerosis. AB - Data, initially anecdotal, but recently supported on more solid experimental evidence, suggest that cannabinoids might be beneficial in the treatment of some of the symptoms of multiple sclerosis (MS). Despite this evidence, there are no data on the possible changes in cannabinoid CB(1) or CB(2) receptors, the main molecular targets for the action of cannabinoids, either in the postmortem brain of patients with MS or in animal models of this disease. The present study addressed this question using the model of experimental allergic encephalomyelitis (EAE) in Lewis rats generated by inoculation of guinea pig myelin basic protein in Freund's adjuvant. After inoculation, animals were examined daily to detect the appearance of neurological signs. The first signs appeared around day 10 after inoculation, reaching the highest degree by day 13, when animals were sacrificed and their brains removed and used for analysis of CB(1) receptor binding, mRNA levels, and activation of GTP-binding proteins. CB(1) receptor binding and mRNA levels were not affected in EAE rats in brain areas such as the hippocampus, limbic structures, and cerebellum. However, there was a marked decrease in both parameters in the caudate-putamen, both in the lateral and medial parts, although this decrease did not correspond with decreases in binding in the nuclei recipient of striatal output neurons, which suggests that changes in CB(1) receptors are exclusively located in the cell bodies of striatal neurons. In addition, CB(1) receptor binding, but not mRNA levels, also decreased in the cerebral cortex, both in the deep and the superficial layers. The analysis of [(35)S]GTPgammaS binding after activation of CB(1) receptors with WIN55,212-2, a synthetic agonist, revealed that, despite the decrease in the number of CB(1) receptors in EAE rats, these were more efficiently coupled to GTP-binding protein-mediated signaling mechanisms in both the caudate-putamen and the cerebral cortex of these animals. In summary, these data suggest that the generation of EAE in Lewis rats would be associated with changes in CB(1) receptors in striatal and cortical neurons, which might be related to the alleviation of some motor signs observed after the treatment with cannabinoid receptor agonists in similar models of MS in rodents. PMID- 11391781 TI - Binding characteristics of selective serotonin reuptake inhibitors with relation to emission tomography studies. AB - When developing ligands for emission tomography studies, one of the major obstacles lies in the selection of ligand candidates. A previously unattended factor such as the influence of temperature on candidate ligand affinity is likely to play a role. By use of rat brain homogenates, the binding characteristics of [(3)H]-(S)-citalopram and [(3)H]-(+)-McN5652 and the receptor ligand interaction at the serotonin transporter of 17 selective serotonin reuptake inhibitors were compared at 21 degrees C and 37 degrees C, respectively. Ligand logP values were also calculated. The ratios for K(i) at 37 degrees C vs. 21 degrees C varied between 0.2 and 2.2 for the selective serotonin reuptake inhibitors considered in this study, with most of the ligands displaying an inverse relationship between K(i) and temperature. Ten of the 17 selective serotonin reuptake inhibitors were found to have pK(i) values statistically significantly different at 21 degrees C compared to 37 degrees C (P < 0.05). The logP values ranged between 3.6 and 4.8, except for DASB, 5-iodo-6-nitroquipazine, and paroxetine where logP was 1.9, 2.2, and 5.0, respectively. K(d) was 0.71 nM at 37 degrees C and 0.31 nM at 21 degrees C for [(3)H]-(S)-citalopram. For [(3)H] (+)-McN5652 K(d) was 0.11 nM at 37 degrees C and 0.08 nM at 21 degrees C. The association and dissociation was much faster for [(3)H]-(S)-citalopram as compared to [(3)H]-(+)-McN5652. It is concluded that temperature may affect K(d) differently and that in vitro dissociation may help to predict whether a given ligand may be useful in PET studies. LogP values do not per se predict the potential of a given ligand as an emission tomography tracer. PMID- 11391782 TI - Loss of metabolites from monkey striatum during PET with FDOPA. AB - The decarboxylation of 6-[(18)F]fluorodopa (FDOPA) and retention of the product [(18)F]fluorodopamine within vesicles of catecholamine fibers results in the labeling of dopamine-rich brain regions during FDOPA/PET studies. However, this metabolic trapping is not irreversible due to the eventual diffusion of [(18)F]fluorodopamine metabolites from brain. Consequently, time-radioactivity recordings of striatum are progressively influenced by metabolite loss. In linear analyses, the net blood-brain clearance of FDOPA (K(D)(i), ml g(-1) min(-1)) can be corrected for this loss by the elimination rate constant k(Lin)(cl) (min(-1)). Similarly, the DOPA decarboxylation rate constant (k(D)(3), min(-1)) calculated by compartmental analysis can also be corrected for metabolite loss by the elimination rate constant k(DA)(9) (min(-1)). To compare the two methods, we calculated the two elimination rate constants using data recorded during 240 min of FDOPA circulation in normal monkeys and in monkeys with unilateral 1-methyl-4 phenyl-1,2,3,6-tetrahydropyridine (MPTP) lesions. Use of the extended models increased the magnitudes of K(D)(i) and k(D)(3) in striatum; in the case of k(D)(3), variance of the estimate was substantially improved upon correction for metabolite loss. The rate constants for metabolite loss were higher in MPTP lesioned monkey striatum than in normal striatum. The high correlation between individual estimates of k(Lin)(cl) and k(DA)(9) suggests that both rate constants reveal loss of decarboxylated metabolites from brain. PMID- 11391783 TI - Gamma-vinyl GABA (GVG) blocks expression of the conditioned place preference response to heroin in rats. AB - We examined the effect of gamma-vinyl GABA (GVG) on the expression of the conditioned place preference response to intraperitoneally (i.p.) administered heroin in rats. Heroin, but not vehicle, produced a significant conditioned place preference response. Pretreatment of animals with 300 mg/kg of GVG significantly attenuated the expression of the heroin-induced conditioned place preference response. These results are the first to suggest that systemic GVG may provide an effective alternative to methadone maintenance in the treatment of heroin addiction, since it is without abuse potential and can be used for treatment outside an institutional setting. PMID- 11391784 TI - Actinomycin D binding to unstructured, single-stranded DNA. AB - Actinomycin D is an anticancer antibiotic best know for inhibiting transcription by binding double-stranded DNA. Tight, sequence selective binding of actinomycin to single-stranded DNA is also known, however, and is implicated in biological activities including inhibition of (-) strand transfer by HIV reverse transcriptase. Oligonucleotide d(GTTAACCATAG) is one of the rare single-stranded DNAs that lack GC steps yet have high affinity for actinomycin. Oligonucleotide sequence and length requirements for drug binding were investigated by monitoring association of the fluorescent surrogate, 7-aminoactinomycin D, to d(GTTAACCATAG) and 31 related oligomers. The TAG-3' terminal sequence was essential for high affinity binding, but was not sufficient. Five oligomers with TAG sequences on or near the 3'-end had high affinity [K(d) < or = 200 nM (oligomer)]. A sixth oligomer, d(GTAACCATATG), had moderately lower affinity (Kd = 370 nM), and other homologous oligomers had much lower affinity. The minimum length sequence for tight binding of 7-aminoactinomycin D was identified as only eight nucleotides, corresponding to d(AACCATAG). This octanucleotide is unstructured in the absence of actinomycin, and has the highest drug affinity of all oligomers examined (Kd = 125 nM). These studies show that high-affinity binding of 7-aminoactinomycin, and actinomycin D by extension, to single-stranded DNA does not require pre-existing secondary structure or any apparent propensity for secondary structure. It is proposed that actinomycin D binds to certain single-stranded DNA sequences by an induced-fit mechanism favored by participation of at least eight nucleotides, or the equivalent of four base pairs. PMID- 11391785 TI - Molecularly imprinted polymers from nicotinamide and its positional isomers. AB - Imprinted polymers were prepared for nicotinamide and its positional isomers. The influence of porogenic solvent and functional monomer on recognition properties of the polymer was compared. The results indicated that two functional groups, the heterocyclic nitrogen and the amide group, in the nicotinamide or isonicotinamide molecule have a synergistic effect in binding to the polymer. The polymers prepared with nicotinamide and isonicotinamide can be used as HPLC stationary phase for the separation of positional isomers of nicotinamide or isonicotinamide, while the polymer prepared with picolinamide showed no specificity toward the template. The mechanisms for the differences in recognition are discussed. In addition to the retention of polymers to their templates the polymers also displayed excellent retention to nicotinic acid and isonicotinic acid, compounds structurally similar to the template. This dual recognition property of the polymer may be useful in circumstances where the preparation of a polymer for a specific template may be problematic because of poor stability or solubility. PMID- 11391786 TI - Effects of tail-like substituents on the binding of competitive inhibitors to the Q(B) site of photosystem II. AB - The QB quinone-binding site of photosystem II is an important target for herbicides. Two major classes of herbicides are based on s-triazine and phenylurea moieties. A small library of triazine and phenylurea compounds has been synthesized which have tail-like substituents in order to test the effects of charge, hydrophobicity and size of the tail on binding properties. It is found that a tail can be attached to one of the alkylamino groups of triazine-type herbicides or to the para position of phenylurea-type herbicides without loss of binding, provided that the tail is hydrophobic. This indicates that the herbicides must be oriented in the QB site such that these positions point toward the natural isoprenyl tail-binding pocket that extends out of the Q(B) site. In turn, the requirement that the tail must extend out of the QB site constrains the size of the other herbicide substituents in the pocket. This is in agreement with the presumed orientation and fit of ligands in the QB site. When longer hydrophobic tails are used, the binding penalty that occurs upon adding a charged substituent at the distal end is reduced. This allows the use of a series of tail substituents possessing a distal charge as an approximate molecular ruler to measure the distance from the QB site to the aqueous phase. Even a 10-carbon alkyl chain still shows a 4-fold effect from the presence or absence of a distal charge. Such a chain does not appear to be long enough to extend from the bulk aqueous phase to the QB site because binding is completely lost when a large hydrophilic domain (PEG(4000)) is attached to the distal end. Longer tails are effective only if they are sufficiently hydrophobic. An effort was made to use tailed herbicides for affinity binding of photosystem II. It was found that hydrophobic linkers promote nonspecific binding, but careful choice of solvent conditions, such as the use of excess nonionic detergent well above its critical micelle concentration, might obviate this problem during affinity-binding applications. PMID- 11391787 TI - NMR chemical shift mapping of the binding site of a protein proteinase inhibitor: changes in the (1)H, (13)C and (15)N NMR chemical shifts of turkey ovomucoid third domain upon binding to bovine chymotrypsin A(alpha). AB - The substrate-like inhibition of serine proteinases by avian ovomucoid domains has provided an excellent model for protein inhibitor-proteinase interactions of the standard type. 1H,15N and 13C NMR studies have been undertaken on complexes formed between turkey ovomucoid third domain (OMTKY3)2 and chymotrypsin A(alpha) (Ctr) in order to characterize structural changes occurring in the Ctr binding site of OMTKY3. 15N and 13C were incorporated uniformly into OMTKY3, allowing backbone resonances to be assigned for OMTKY3 in both its free and complex states. Chemical shift perturbation mapping indicates that the two regions, K13 P22 and N33-A40, are the primary sites in OMTKY3 involved in Ctr binding, in full agreement with the 12 consensus proteinase-contact residues of OMTKY3 defined previously on the basis of X-ray crystallographic and mutational analysis. Smaller chemical shift perturbations in selected other regions may result from minor structural changes on binding. Through-bond 15N-13C correlations between P1 13C' and P1'-15N in two-dimensional H(N)CO and HN(CO) NMR spectra of selectively labeled OMTKY3 complexed with Ctr indicate that the scissile peptide bond between L18 and E19 of the inhibitor is intact in the complex. The chemical shifts of the reactive site peptide bond indicate that it is predominantly trigonal, although the data are not inconsistent with a slight perturbation of the hybridization of the peptide bond toward the first tetrahedral state along the reaction coordinate. PMID- 11391788 TI - Docking of combinatorial peptide libraries into a broadly cross-reactive human IgM. AB - A monoclonal IgM cryoglobulin with diverse binding behavior was isolated from a patient (Mez) with Waldenstrom's macroglobulinemia. It gave very high titers in the binding of combinatorially synthesized libraries of peptides ranging in size from two to eight residues. The crystal structure of Mez Fv revealed that the binding site was divided into two cavities of unequal volumes with dimensions and chemical properties that were compatible with the binding of peptides. Access to this unique combination of structural information and peptide binding data led us to carry out Mez-peptide docking simulations to gain insight into the Mez binding propensities. In the present article, the results for docking of five peptide libraries are combined with discussions of the methods and approximations involved in the docking process. We analyze the origins of peptide binding affinity for Mez IgM in terms of its cross-reactivity and its structural preferences. PMID- 11391789 TI - Optical biosensor investigation of interactions of biomembrane and water-soluble cytochromes P450 and their redox partners with covalently immobilized phosphatidylethanolamine layers. AB - A phospholipid-containing biochip was created by covalently immobilizing phospholipids on the optical biosensor's aminosilane cuvette and employed to monitor the interactions of the membrane and water-soluble proteins in cytochrome P450-containing monooxygenase systems with planary layers of dilauroylphosphatidylethanolamine (DLPE) and distearoylphosphatidylethanolamine (DSPE), differing in acyl chain length. It was shown that the full-length membrane proteins-cytochrome P4502B4 (d-2B4), cytochrome b5 (d-b5) and NADPH cytochrome P450 reductase (d-Fp)-readily incorporated into the phospholipids. The incorporation was largely due to hydrophobic interactions of membranous protein fragments with the phospholipid layer. However, electrostatic forces were also but not always involved in the incorporation process. They promoted d-Fp incorporation but had no effect on d-b5 incorporation. In low ionic strength buffer, no incorporation of these two proteins into the DSPE lipid layer was observable. Incorporation of d-b5 into the DLPE layer was abruptly increased at temperatures exceeding phospholipid phase transition point. Incorporation of d 2B4 was dependent on its aggregation state and decreased with increasing protein aggregability. Water-soluble proteins either would not interact with the phospholipid layer (adrenodoxin) or would bind to the layer at the cost of only electrostatic (albumin) or both electrostatic and hydrophobic (P450cam) interactions. PMID- 11391790 TI - Detection of unidentified chromosome abnormalities in human neuroblastoma by spectral karyotyping (SKY). AB - Spectral karyotyping (SKY) is a novel technique based on the simultaneous hybridization of 24 fluorescently labeled chromosome painting probes. It provides a valuable addition to the investigation of many tumors that can be difficult to define by conventional banding techniques. One such tumor is neuroblastoma, which is often characterized by poor chromosome morphology and complex karyotypes. Ten primary neuroblastoma tumor samples initially analyzed by G-banding were analyzed by SKY. In 8/10 tumors, we were able to obtain additional cytogenetic information. This included the identification of complex rearrangements and material of previously unknown origin. Structurally rearranged chromosomes can be identified even in highly condensed metaphase chromosomes. Following the SKY results, the G-banding findings were reevaluated, and the combination of the two techniques resulted in a more accurate karyotype. This combination allows identification not only of material gained and lost, but also of breakpoints and chromosomal associations. The use of SKY is therefore a powerful tool in the genetic characterization of neuroblastoma and can contribute to a better understanding of the molecular events associated with this tumor. PMID- 11391791 TI - Cyclin D1 gene contains a cryptic promoter that is functional in human cancer cells. AB - A novel cyclin D1 (CCND1)-TROP2 fusion oncogene has been isolated from human cancer cells. Unexpectedly, the chimeric cDNA was found to express TROP2 in the absence of exogenous promoters. Mutagenesis of the TROP2 and CCND1 sequences and in vitro transcription/translation show that a cryptic promoter is present in the 3' coding region of CCND1. The CCND1 cryptic promoter is functional in luciferase assays, where it augments the basal expression levels by eightfold and efficiently cooperates with an SV-40 enhancer. The transcription start sites of the cryptic promoter map at bases 797 and 935 of CCND1, as determined by RNase protection assays. The cryptic promoter possesses canonical binding sites for ubiquitous transcription factors and W/S, X1, and CAAT/Y boxes that are characteristic of major histocompatibility complex class II gene promoters. Remarkably, the cryptic CCND1 promoter is active in human cancer cells and generates a truncated transcript that contains CCND1 instability sequences. Thus, this novel CCND1 transcription unit may play a role in the regulation of the expression of cyclin D1 and in tumor cell growth. PMID- 11391792 TI - Allelic loss and gain, but not genomic instability, as the major somatic mutation in primary hepatocellular carcinoma. AB - To identify genetic abnormalities in primary hepatocellular carcinoma (HCC), we performed microsatellite analysis (MSA) on 60 Chinese HCC specimens. Utilizing a semi-quantitative MSA and 292 highly polymorphic markers spanning all 22 autosomes, we found that somatic allelic imbalance (AI) occurred frequently in HCC. To evaluate the nature of the AI, comparative genomic hybridization was performed on 20 HCC specimens. The combined use of these two methods revealed frequent allelic loss on 17p, 9p21-p23, 4q, 16q21-q23.3, 13q, 8p21-p23, and 6q24 q27, whereas there was frequent allelic gain on 1q, 17q, and 8q24. The region with the highest incidence of genomic imbalance was 17p13 (65%), followed by 9p21 p23 (55%), 4q (35-51%), 16q21-q23.3 (52%), 17p12 (49%), 13q (39-46%), 8p21-p23 (41-45%), 8q24 (41%), and 1q32 (40%). In addition, aberrations of 19p13.3, 16p13.3, 13q33-q34, 9q13-31, and 7q were reported for the first time. The presence of a close correlation of 17p13 deletion with abnormalities of some other loci implies that 17p13 could play a crucial role in oncogenesis. Interestingly, microsatellite instability was rarely seen in our patients, in contrast to that observed in European HCC samples. PMID- 11391793 TI - Smallest region of overlapping deletion in 1p36 in human neuroblastoma: a 1 Mbp cosmid and PAC contig. AB - In human neuroblastomas, the distal portion of 1p is frequently deleted, as if one or more tumor suppressor genes from this region were involved in neuroblastoma tumorigenesis. Earlier studies had identified a smallest region of overlapping deletion (SRO) spanning approximately 23 cM between the most distally retained D1S80 and by the proximally retained D1S244. In pursuit of generating a refined delineation of the minimally deleted region, we have analyzed 49 neuroblastomas of different stages for loss of heterozygosity (LOH) from 1pter to 1p35 by employing 26 simple sequence length polymorphisms. Fifteen of the 49 tumors (31%) had LOH; homozygous deletion was not detected. Seven tumors had LOH at all informative loci analyzed, and eight tumors showed a terminal or an interstitial allelic loss of 1p. One small terminal and one interstitial deletion defined a new 1.7 cM SRO, approximately 1 Mbp in physical length, deleted in all tumors between the retained D1S2731 (distal) and D1S2666 (proximal). To determine the genomic complexity of the deleted region shared among tumors, we assembled a physical map of the I Mbp SRO consisting predominantly of bacteriophage P1 derived artificial chromosome (PAC) clones. A total of 55 sequence-tagged site (STS) markers (23 published STSs and short tandem repeats and 32 newly identified STSs from the insert ends of PACs and cosmids) were assembled in a contig, resulting in a sequence-ready physical map with approximately one STS per 20 Kbp. Twelve genes (41BB, CD30, DFFA, DJ1, DR3, FRAP, HKR3, MASP2, MTHFR, RIZ, TNR2, TP73) previously mapped to 1p36 are localized outside this SRO. On the basis of this study, they would be excluded as candidate genes for neuroblastoma tumorigenesis. Ten expressed sequence tags were integrated in the contig, of which five are located outside the SRO. The other five from within the SRO may provide an entrance point for the cloning of candidate genes for neuroblastoma. PMID- 11391794 TI - Combined copy status of 18q21 genes in colorectal cancer shows frequent retention of SMAD7. AB - Deletions of chromosome band 18q21 appear with very high frequency in a variety of carcinomas, especially in colorectal cancer. Potent tumor suppressor genes located in this region encode transforming growth factor beta (TGF-beta) signal transducers SMAD2 and SMAD4, and inactivation of either one leads to impaired TGF beta-mediated cell growth/apoptosis. Following the assignment of SMAD7 to 18q21, we first refined the SMAD7 gene position within this region by genetically mapping SMAD7 between SMAD2 and SMAD4. Further, to compare the respective frequencies of genetic alterations of these three SMAD genes in colorectal cancer, we undertook a large-scale evaluation of the copy status of each of these genes on DNA samples from colorectal tumor biopsy material. Among a subset of 233 DNA samples for which data were available for all four genes, SMAD4, SMAD2, and the nearby gene DCC showed high deletion rates (66%, 64%, and 59%, respectively), whereas SMAD7 was deleted in only 48% of the tumors. Unexpectedly, we found some gene duplications; SMAD7 appears to be more frequently amplified (10%) than the three other genes (4-7%). Compiled data for SMAD genes in each tumor show that the most common combination (26% of all the tumors) consists of the simultaneous deletions of SMAD2 and SMAD4 associated with normal diploidy or even duplication of SMAD7. Since SMAD7 normally counteracts SMAD2 and SMAD4 in TGF-beta signaling, we hypothesize that the tumor might not benefit from simultaneous SMAD7 inactivation, thereby exerting selective pressure to retain or even to duplicate the SMAD7 gene. PMID- 11391795 TI - A complex pattern of recurrent chromosomal losses and gains in T-cell prolymphocytic leukemia. AB - T-cell prolymphocytic leukemia (T-PLL) is a rare malignant proliferation of lymphoid cells with a postthymic phenotype. Previous cytogenetic and molecular studies reported complex karyotypes with recurrent chromosomal abnormalities, including translocations involving either TCL1 at 14q32.1 or MTCP1 at Xq28, inactivation of the ATM gene by deletion and/or mutation, and isochromosomes 8. For extensive study of chromosomal imbalances in T-PLL, we analyzed 22 tumoral DNAs using comparative genomic hybridization (CGH). Abnormal CGH profiles were detected in all cases, demonstrating highly recurrent gains and losses and largely extending the abnormalities previously established. Only a few nonrecurrent abnormalities were observed, in contrast to the genetic instability anticipated from ATM inactivation. Nine recurrent regions of loss were identified at 8p (frequency 86%), 11q (68%), 22q11 (45%), 13q (41%), 6q (36%), 9p (27%), 12p (23%), 11p11-p14 (23%), and 17p (23%), as well as four regions of gain at 8q (82%), 14q32 (50%), 22q21-qter (41%), and 6p (23%). Several recurrent chromosomal abnormalities were simultaneously present in each case (mean, 5.7; up to 10), none being mutually exclusive of another. Fluorescence in situ hybridization analysis confirmed and extended 22q11 and 13q losses, giving final frequencies of 55% and 45%, respectively. Analysis of one case over a 7-year period confirmed the overall genetic stability of T-PLL and showed that tumor progression was associated with the onset of a few chromosomal abnormalities. This study establishes a complex pattern of highly recurrent chromosomal abnormalities in T PLL, including some, such as chromosome 13 deletion, commonly found in other lymphoid malignancies. PMID- 11391796 TI - Novel mutations in the SDHD gene in pedigrees with familial carotid body paraganglioma and sensorineural hearing loss. AB - Paraganglioma (PGL) is a rare disorder characterized by tumors of the head and neck region. Between 10% and 50% of cases of PGL are familial, and the disease is autosomal dominant and subject to age-dependent penetrance and imprinting. The paraganglioma gene (PGL1) has been mapped to 11q22.3-q23, and recently germline mutations in the SDHD gene have been identified. The SDHD region contains another gene, DPP2/TIMM8B, the homolog of which causes dystonia and deafness seen in Mohr Tranebjaerg syndrome. Using four PGL pedigrees, two of which exhibit coinheritance of PGL and sensorineural hearing loss or tinnitus, analysis of 14 microsatellite markers provided support for linkage to the PGL1 locus. Sequence analysis identified novel mutations in exon 1 and exon 3 of the SDHD gene, including a novel two base pair deletion in exon 3 creating a premature stop codon at position 67; a novel three base pair deletion in exon 3 resulting in the loss of Tyr-93; a missense mutation in exon 3 resulting in the substitution of Leu-81 for Pro-81; and a novel G-to-C substitution in exon 1 resulting in the substitution of Met-1 for Ile-1. No base changes were detected in the DPP2/TIMM8B gene. There was no apparent loss of heterozygosity at the site of the SDHD mutations. However, RT-PCR analysis of tumor samples showed monoallelic expression of the mutant (paternal) allele as expected for imprinting. This has not previously been shown for this disorder. The inheritance and expression of the SDHD gene is consistent with the PGL1 gene being subject to genomic imprinting. PMID- 11391797 TI - Ectopic sequences from truncated HMGIC in liposarcomas are derived from various amplified chromosomal regions. AB - The HMGIC gene codes for an architectural transcription factor frequently rearranged by translocation in lipomas and other benign mesenchymal tumors. In sarcomas, malignant tumors of mesenchymal origin, the gene is also found to be rearranged, but in addition amplified and overexpressed. Here we report the sequence, chromosomal localization, and expression patterns of 11 novel ectopic sequences fused to exons 2 and 3 of HMGIC in seven different sarcoma samples. In addition, we identified a number of variant transcripts observed previously in benign tumors. Consistent with the suggested role of HMGIC in adipocytic differentiation, most of the novel ectopic sequences were observed in well differentiated liposarcomas. These tumors are known to have complex marker chromosomes containing amplified segments from several chromosomes. Five novel sequences were derived from 12q14-q15, where HMGIC resides, two from 1q24, a region frequently amplified in these types of tumors, two from 11q14, and one from chromosome 2. All except one of the aberrant transcripts encoded truncated proteins with intact DNA-binding domains (AT hooks) but lacking the C-terminal acidic region, a target for constitutive phosphorylation by protein kinase CK2. Some of the ectopic sequences were transcribed in other tissues, and most of the ectopic sequences also showed recurrent amplification in liposarcomas. PMID- 11391798 TI - Nearly all hereditary paragangliomas in the Netherlands are caused by two founder mutations in the SDHD gene. AB - Hereditary paragangliomas or glomus tumors are usually benign slow-growing tumors in the head and neck region. The inheritance pattern of hereditary paraganglioma is autosomal dominant with imprinting. Recently, we have identified the SDHD gene encoding subunit D of the mitochondrial respiratory chain complex II as one of the genes involved in hereditary paragangliomas. Here, we demonstrate that two founder mutations, Asp92Tyr and Leu139Pro, are responsible for paragangliomas in 24 and 6 of the 32 independently ascertained Dutch paraganglioma families, respectively. These two mutations were also detected among 20 of 55 isolated patients. Ten of the isolated patients had multiple paragangliomas, and in eight of these SDHD germline mutations were found, indicating that multicentricity is a strong predictive factor for the hereditary nature of the disorder in isolated patients. In addition, we demonstrate that the maternally derived wild-type SDHD allele is lost in tumors from mutation-carrying patients, indicating that SDHD functions as a tumor suppressor gene. PMID- 11391799 TI - Genomic imbalances in human lung adenocarcinomas and squamous cell carcinomas. AB - Comparative genomic hybridization analysis was performed on 67 non-small-cell lung cancers (NSCLCs), including 32 squamous cell carcinomas (SCCs) and 35 adenocarcinomas (ACs), to identify differences in the patterns of genomic imbalance between these two histologic subtypes. Among the entire tumor set, the chromosome arms most often overrepresented were 1q, 3q, 5p, and 8q, each detected in 50-55% of cases. The most frequently underrepresented arms were 9q, 3p, 8p, and 17p. The number of imbalances was similar in SCCs and ACs (median number/case: 12 and 11, respectively). Moreover, many imbalances, such as gains of 1q, 5p, and 8q, occurred at a high frequency in both histologic subgroups. Several statistically significant differences, however, were found. The most prominent difference was gain of 3q24-qter, seen in 81% of SCCs compared with 31% of ACs (P < 0.0001), with amplification at 3q25-26 being detected in eight of 32 (25%) SCCs but in only two of 35 (6%) ACs. Gain of 20p13 and loss of 4q also were seen at a significantly higher rate in SCCs than in ACs, whereas overrepresentation of 6p was more common in ACs. Gains of 7q and 8q each were associated with higher-stage tumors and either positive nodal involvement or higher tumor grade. These data suggest that genes located in several chromosomal regions, particularly 3q25-26, may be associated with phenotypic properties that differentiate lung SCCs from ACs. Furthermore, certain imbalances, prominent among them gains of 7q and 8q, may be indicative of tumor aggressiveness in NSCLCs. PMID- 11391800 TI - Novel ERBB4 juxtamembrane splice variants are frequently expressed in childhood medulloblastoma. AB - We recently reported a significant relationship between tumor cell expression of the ERBB4 receptor, the most recently described member of the epidermal growth factor receptor family, and aggressive tumor phenotype in childhood medulloblastoma. Two alternative juxtamembrane (JM) isoforms of the ERBB4 receptor have been described. Termed JMa and JMb, these variants possess different receptor processing and ligand-binding characteristics. In the current study, we employed an RT-PCR and sequencing strategy to determine the pattern of ERBB4 JM isoform expression in a large (n = 78) series of pediatric medulloblastomas. JMa and JMb transcript expression was detected in 53% and 28% of tumor samples, respectively. In addition, two novel ERBB4 JM isoforms, which we have termed JMc and JMd, were isolated from 10% and 36% of tumors, respectively. Sequence analysis revealed the JMc transcript to contain a deletion of the entire JM region. In contrast, JMd includes an extended coding region, retaining both the JMa and JMb sequences. Neither of these novel isoforms was detected in normal human adult cerebellum, but expression of JMd was observed in developing fetal cerebellum, suggesting that this later isoform may represent an ERBB4 transcript restricted to primitive neuroectoderm-derived tissue. To confirm that the four ERBB4 JM isoforms arise by alternative RNA splicing, we sequenced the intron-exon junctions of the human ERBB4 gene within the JM region. This demonstrated the four ERBB4 JM variants to be encoded by two short exons containing the JMb and JMa sequences positioned in the order 5' to 3' and separated by a 121 bp intron. PMID- 11391802 TI - Heterogeneous tumor clones as an explanation of discordance between plasma DNA and tumor DNA alterations. PMID- 11391801 TI - Exclusion of SMAD4 mutation as an early genetic change in human pancreatic ductal tumorigenesis. AB - Pancreatic ductal carcinoma is one of the malignant diseases with the poorest prognosis. To develop effective methods for better treatment of pancreatic cancer patients, we tried to analyze the course of multistep carcinogenesis of the pancreatic ductal cells. IPMT (intraductal papillary-mucinous tumor) is thought to be one of the premalignant lesions of the pancreas, which would transform into carcinomas. Loss of 18q at the SMAD4 locus is known to be an early genetic change in pancreatic ductal carcinomas. It is not clear, however, whether or not the target gene for inactivation is SMAD4. Using 18 IPMTs, we analyzed LOH at the SMAD4 locus and observed frequent LOH (7/14, 50%). No mutations were observed in any of the tumors. Moreover, the expression level of the SMAD4 protein did not show a reduction in IPMTs. These results suggested that (i) inactivating mutation of the SMAD4 gene is a rather late genetic change in pancreatic carcinogenesis, and (ii) there may be an unknown tumor suppressor gene in 18q, other than SMAD4, that is involved in pancreatic ductal carcinogenesis. PMID- 11391803 TI - Kinetic energy release distributions in mass spectrometry. AB - Kinetic energy releases (KERs) in unimolecular fragmentations of singly and multiply charged ions provide information concerning ion structures, reaction energetics and dynamics. This topic is reviewed covering both early and more recent developments. The subtopics discussed are as follows: (1) introduction and historical background; (2) ion dissociation and kinetic energy release: kinematics; potential energy surfaces; (3) the kinetic energy release distribution (KERD); (4) metastable peak observations: measurements on magnetic sector and time-of-flight instruments; energy selected results by photoelectron photoion coincidence (PEPICO); (5) extracting KERDs from metastable peak shapes; (6) ion structure determination and reaction mechanisms: singly and multiply charged ions; biomolecules and fullerenes; (7) theoretical approaches: phase space theory (PST), orbiting transition state (OTS)/PST, finite heat bath theory (FHBT) and the maximum entropy method; (8) exit channel interactions; (9) general trends: time and energy dependences; (10) thermochemistry: organometallic reactions, proton-bound clusters, fullerenes; and (11) the efficiency of phase space sampling. PMID- 11391804 TI - Matrix-assisted laser desorption/ionization Fourier transform mass spectrometric analysis of oxygenated triglycerides and phosphatidylcholines in egg tempera paint dosimeters used for environmental monitoring of museum display conditions. AB - Oxidative changes in triacylglycerols and diacylphosphatidylcholines in egg tempera paint strips are used for chemical dosimetry of the quality of the museum environment. High-resolution matrix-assisted laser desorption/ionization Fourier transform mass spectrometry (MALDI-FTMS) was used as a rapid method for the determination of the exact elemental composition of the alteration products from diacylphosphatidylcholines and triacylglycerols. Light exposure of the egg tempera paints yields oxygenated diacylphosphatidylcholines and triacylglycerols. In the latter multiple incorporation of oxygen was observed as a recurring mass difference of 15.995, the exact atomic mass of oxygen. Owing to the high resolution of the FTMS data (routinely 20 000 at m/z 1000 in broadband mode), oxidation products with different elemental compositions but identical nominal mass could be distinguished. Products of oxidative cleavage of triacylglycerols were observed in samples exposed for longer times. The relative intensities of the peaks of singly and multiply oxygenated triacylglycerols were used to derive the degree of oxygenation of the egg lipids in the tempera paint dosimeters. The degree of oxygenation was found to be directly related to the light exposure time. Exposure to elevated temperature (60 degrees C) for a period of 21 days did not lead to oxygenation of the triacylglycerols and diacylphosphatidylcholines. Exposure to NO(x) and SO(2) in the dark greatly increased the degree of oxygenation. Addition of lead- or copper-containing pigments to the egg binding medium (and subsequent storage for 6 months in the dark) led to accelerated conversion of egg lipids to oxidised products. PMID- 11391805 TI - Formation and photodissociation of M(+)-C(6)H(6) (M(+) = V(+) and Ta(+)) and Ta(+)-C(6)H(4) complexes in a time-of-flight mass spectrometer. AB - A series of cyclic hydrocarbons were introduced to react with V(+) and Ta(+) using a pulsed beam expansion source in a time-of-flight mass spectrometer. The third-row metal Ta(+) displayed high reactivity in dehydrogenation to form benzyne complexes, whereas benzene complexes were the terminal products for V(+). M(+)-C(6)H(6) (M(+) = V(+) and Ta(+)) and Ta(+)-C(6)H(4) were selected to perform the photodissociation experiments. In contrast to the V(+) fragment formation via simple cleavage of the V(+)-C(6)H(6) bond, a photoinduced loss of C(2)H(2) occurred in both the Ta(+)-C(6)H(6) and Ta(+)-C(6)H(4) complexes. Plausible explanations involved in the formation of Ta(+)-C(6)H(6) and Ta(+)-C(6)H(4) complexes are given for observing such photo-induced dissociation. The observed photodissociation in Ta(+)-C(6)H(6) is analogous to the dissociative process previously investigated in metal ion-molecule reactions. The photodissociation spectrum of Ta(+)-C(6)H(4) was obtained by recording the appearance of Ta(+) C(4)H(2) as a function of wavelength and yielded a dissociation energy of 91 +/- 1 kcal mol(-1). PMID- 11391807 TI - Electrospray ionization tandem mass spectrometric study of salt cluster ions: part 2--salts of polyatomic acid groups and of multivalent metals. AB - Salt cluster ions formed from 0.05 M solutions of CaCl(2), CuCl(2) and Na(A)B (where A = 1 or 2 and B = CO(3)(2-), HCO(3)(-), H(2)PO(4)(-) and HPO(4)(2-)) were studied by electrospray ionization tandem mass spectrometry. The effects on salt cluster ions of droplet pH and of redox reactions induced by electrospray provide information on the electrospray process. CaCl(2) solution yielded salt cluster ions of the form (CaCl(2))(n)(CaCl)(x)(x+) and (CaCl(2))(n)(Cl)(y)(y-), where x, y = 1-3, in positive- and negative-ion modes, respectively. Upon collision induced dissociation (CID), singly charged CaCl(2) cluster ions fragmented, doubly charged cluster ions generated either singly or both singly and doubly charged fragment ions, depending on the cluster mass, and triply charged clusters fragmented predominantly by the loss of charged species. CuCl(2) solution yielded nine series of cluster ions of the form (CuCl(2))(n)(CuCl)(m) plus Cu(+), CuCl(+), or Cl(-). CuCl, the reductive product of CuCl(2), was observed as a neutral component of positively and negatively charged cluster ions. Free electrons were formed in a visible discharge that bridged the gap between the electrospray capillary and the sampling cone brought about the reduction of Cu(2+) to Cu(+). Upon CID, these cluster ions fragmented to lose CuCl(2), CuCl, Cl, and Cl(2). Na(2)CO(3) and NaHCO(3) solutions yielded cluster ions of the form (Na(2)CO(3))(n) plus Na(+) or NaCO(3)(-). Small numbers of NaHCO(3) molecules were found in some cluster ions obtained with the NaHCO(3) solution. For both Na(2)HPO(4) and NaH(2)PO(4) solutions, ions of the form (Na(2)HPO(4))(h), (NaH(2)PO(4))(i), (Na(3)PO(4))(j), (NaPO(3))(k) plus Na(+), PO(3)(-) or H(2)PO(4)(-) were observed. In addition, ions having one or two phosphoric acid (H(3)PO(4)) molecules were observed from the NaH(2)PO(4) solution while ions containing one sodium hydroxide (NaOH) molecule were observed from the Na(2)HPO(4) solution. The cluster ions observed from these four salts of polyatomic acid groups indicate that changes in pH occur in both directions during the electrospray process principally by solvent evaporation; the pH value of the acidic solution became lower and that of the basic solution higher. PMID- 11391806 TI - Mass spectrometric studies of the path of carbon in photosynthesis: positional isotopic analysis of (13)C-labelled C(4) to C(7) sugar phosphates. AB - The (EIMS) electron ionization mass spectrometric fragmentation patterns of the methoxime- and ethoxime-trimethylsilyl (TMS) derivatives of C(4) to C(7) sugars involved as phosphates in the Calvin pathway of photosynthesis in plants were analysed by gas chromatography/EIMS using specifically labelled (13)C analogs. In general, most but not all of the major ions in the mass spectra arise from single carbon-carbon bond cleavages of the straight-chain derivatives. The results confirm that GC/MS of the alkoxime-TMS derivatives is a viable method for measuring (13)C incorporations at individual carbon atoms in each of the sugar phosphates during photosynthetic experiments with (13)CO(2). PMID- 11391808 TI - Field desorption mass spectrometry of large multiply branched saturated hydrocarbons. AB - Large multiply branched saturated hydrocarbons containing 67-103 carbon atoms (molecular masses 941.8-1446.8 Da) were analyzed by field desorption mass spectrometry (FD-MS) with a double-focusing mass spectrometer. FD-MS was found to have detection limits in the 100 fmol range. The FD mass spectra exhibited molecular ions of astonishingly low abundance. However, the fragment ions formed were closely related to the proposed molecular structure, allowing us to set up rules for straightforward structure elucidation of unknowns. In detail, (i) dehydrogenation, (ii) alkyl losses from molecular ions and (iii) subsequent alkene losses were observed. The influence of the electric field strength on dehydrogenation and C-C cleavages was examined by variation of the emitter potential. Additionally, ion dissociations in the ion source and in the first and second field-free regions, respectively, were compared to study the relative importance of field-induced and thermally induced processes. PMID- 11391809 TI - Characterization and differentiation of ruthenium(II) complexes with 1,4,7 trithiacyclononane and nitrogen heterocycles by electrospray mass spectrometry. AB - Electrospray ionization mass spectrometry (ESIMS) was used to characterize a series of new ruthenium(II) complexes with several nitrogen heterocycles and a common ligand: the crown thioether 1,4,7-trithiacyclononane, [9]aneS(3). ESIMS allows the easy identification of the [Ru(II)Cl([9]aneS(3))Y]X complexes, where Y is a bidentate nitrogen heterocycle and X is Cl(-) or PF(6)(-), through the formation of two diagnostic ions by fragmentation of the common ligand [9]aneS(3). Structures for these ions and mechanisms for their gas-phase formation are proposed based on data from product ion spectra. PMID- 11391811 TI - Determination of polychlorodibenzo-p-dioxins and polychlorodibenzofurans by gas chromatography/tandem mass spectrometry and gas chromatography/triple mass spectrometry in a quadrupole ion trap. AB - The determination of tetra- to octachlorodibenzo-p-dioxins and tetra- to octachlorodibenzofurans (PCCD/Fs) by high-resolution gas chromatography/tandem mass spectrometry (HRGC/MS/MS) and high-resolution gas chromatography/triple mass spectrometry (HRGC/MS(3)) in a quadrupole ion trap, equipped with an external ion source, is presented. MS/MS involves a typical four-step process, namely ionization, parent ion isolation, collision-induced dissociation (CID) and mass analysis of the daughter ions. For the MS(3) experiment, the MS/MS scan function is used with the addition of selected daughter ion isolation, their CID and the mass analysis of second-generation product ions called 'grand-daughter ions.' For both methods, the energies necessary for the CID of the 17 PCDD/Fs were determined and optimized using multiple scan functions with different CID amplitudes. The CID efficiency, defined as the signal ratio of fragment ions detected from the major dissociation channels to molecular ions isolated, was 1.15-2.40 V for parent ion dissociation (MS/MS) and 1.05-1.50 V for daughter ion dissociation (MS(3)) and for all the chloro congeners. The same sensitivity (1 pg microl(-1)) can be reached with both the MS/MS and MS(3) methods and linear responses were obtained between 1 and 100 pg microl(-1) injected. PMID- 11391810 TI - Intensity dependence of cation kinetic energies from 2,5-dihydroxybenzoic acid near the infrared matrix-assisted laser desorption/ionization threshold. AB - The mechanisms responsible for matrix-assisted laser desorption/ionization (MALDI) are far from being well understood, particularly where infrared laser irradiation is used to initiate the process. We measured the emission yields and kinetic energy distributions of positive ions emitted from 2,5-dihydroxybenzoic acid loaded with angiotensin II in a standard MALDI preparation during irradiation with an infrared free-electron laser tuned to 2.94 microm. As the laser intensity is scanned through the MALDI threshold, we see a marked change in the energy distributions of the matrix ion. Above threshold, the energy distributions of both analyte and matrix cations are constant over a broad range of laser intensities. This behavior does not appear to be consistent with any extant model of the MALDI mechanism. PMID- 11391812 TI - Mass spectrometric studies of trimethylsilylpantothenic acid and related substances. AB - The characteristic fragment of trimethylsilylated pantothenic acid (TMS-PA) at m/z 291 upon electron ionization was shown to originate from the molecular ion by a McLafferty rearrangement instead of by ejection of 1,1,3,3-tetramethyl-1,3 disilacyclobutane. The verification consisted of labelling experiments and high resolution mass spectrometry of the fragment and studies on its isotopic distribution. The remaining fragmentation pathways of TMS-PA were clarified by B/E-linked scans and collision-induced dissociation. PMID- 11391813 TI - Investigation of different combinations of derivatization, separation methods and electrospray ionization mass spectrometry for standard oligosaccharides and glycans from ovalbumin. AB - Derivatization procedures using 1-phenyl-3-methyl-5-pyrazolone (PMP) and 2 aminonaphthalene trisulfone (ANTS) were selected among a number of well known methods for labelling carbohydrates. PMP derivatives were selected owing to our laboratory's previous high-performance liquid chromatography/electrospray ionization mass spectrometry (HPLC/ESI-MS) experience with these, whereas the ANTS-labelled compounds were prepared for fluorophore-assisted carbohydrate electrophoresis (FACE) separation. ANTS-oligosaccharide standards were characterized to study their ionization patterns. Reversed-phase and normal-phase HPLC systems were coupled on-line with ESI-MS. Each necessitated its own mobile phase system which, in turn, imposed some important changes in the ionization conditions used and/or on the ionization patterns and spectra obtained. Following characterization of the intact glycoprotein ovalbumin with ESI-MS, its glycans were detached using the enzyme PNGase-F. The glycans were subjected to PMP and ANTS derivatization. It was very difficult to separate ANTS derivatives by reversed-phase HPLC owing to lack of retention, and normal-phase HPLC offered reasonable retention with limited separation. PMP compounds overall yielded better normal- and reversed-phase separations and improved sensitivity over the ANTS-labelled sugars, for which negative mode ESI had to be used. The combination of ESI of intact ovalbumin and ESI of PMP-glycans gave rise to the detection of over 20 different glycoforms, excluding the possible presence of structural isomers for each sugar composition detected. PMID- 11391814 TI - Elucidation of fragmentation mechanisms involving transfer of three hydrogen atoms using a quadrupole time-of-flight mass spectrometer. PMID- 11391815 TI - Current awareness. Current literature in mass spectrometry. AB - In order to keep subscribers up-to-date with the latest developments in their field, John Wiley & Sons are providing a current awareness service in each issue of the journal. The bibliography contains newly published material in the field of mass spectrometry. Each bibliography is divided into 11 sections: 1 Books, Reviews & Symposia; 2 Instrumental Techniques & Methods; 3 Gas Phase Ion Chemistry; 4 Biology/Biochemistry: Amino Acids, Peptides & Proteins; Carbohydrates; Lipids; Nucleic Acids; 5 Pharmacology/Toxicology; 6 Natural Products; 7 Analysis of Organic Compounds; 8 Analysis of Inorganics/Organometallics; 9 Surface Analysis; 10 Environmental Analysis; 11 Elemental Analysis. Within each section, articles are listed in alphabetical order with respect to author (3 Weeks journals - Search completed at 7th. Mar. 2001) PMID- 11391816 TI - Presence of proliferating (MiB-1-positive) cells in cervical smears of women infected with HIV is associated with clinical outcome: a study of Brazilian women. AB - The hypothesis tested was that there is an association between the presence of proliferating (MiB-1-positive) cervical cells and clinical outcome of women infected with human immunodeficiency virus (HIV). Female partners (attending the Gynecology Outpatients Clinic of the University Hospital of Rio Grande, Brazil) of known HIV-positive (HIV+) men were used for this pilot study. Among these women, 25 were also HIV+. Papanicolaou smears of these 25 HIV+ women and of 44 HIV- women were graded as negative, CIN I, CIN II, or CIN III, using neural network screening. MiB-1 grading and HPV identification were also performed. The immune status of patients was determined using the current Centers for Disease Control classification. In agreement with the scientific literature, in these Brazilian women both CIN and HPV were associated with HIV. In the HIV+ women, the immune status tends to correlate with MiB-1 grading. Also, in the one case in whom progression from CIN I to invasive cervical carcinoma was observed, the smear contained many MiB-1-positive cells. Staining cervical smears of HIV+ women is a simple procedure to get an indication of clinical outcome of the patient. PMID- 11391817 TI - Morphologic predictors of papillary carcinoma on fine-needle aspiration of thyroid with ThinPrep preparations. AB - Although the cytologic features of papillary carcinoma of the thyroid are well known, none is entirely specific. We conducted this study to determine the minimal criteria necessary to achieve 100% specificity for the diagnosis of papillary carcinoma on fine-needle aspiration (FNA). Forty patients with histologically confirmed papillary carcinoma and 17 patients with other thyroid lesions who underwent preoperative FNA at Beth Israel Deaconess Medical Center during a 4-yr period were included in the study. All cytology slides were prepared with the ThinPrep processing technique. Various architectural and nuclear features were evaluated, with a score assigned to each feature, and correlated with the histologic diagnosis of papillary carcinoma. Intranuclear inclusions, papillary and/or sheet arrangements, nuclear grooves, powdery chromatin, nuclear molding, high cellularity, and small nucleoli were significantly associated with papillary carcinoma (P < 0.05). The requirement of any intranuclear inclusions and many nuclear grooves, or a minimum of sum of scores (of the above eight features) of 10, yields 100% specificity and approximately 70% sensitivity. Cases with fewer features can be reported as suspicious or indeterminate for papillary carcinoma. A quantitative/probabilistic approach in the reporting of thyroid FNA provides a practical guide for management of patients with thyroid nodules. PMID- 11391818 TI - Cytologic spectrum of 227 fine-needle aspiration cases of chest-wall lesions. AB - This study was carried out with the objective of studying the cytomorphology of the wide variety of chest-wall lesions. Two hundred twenty-seven chest-wall lesions were studied over a period of 4 yr. Routine May-Grunwald-Giemsa (MGG) and hematoxylin-eosin (H&E) slides were studied along with special stains, whenever required. The malignant lesions comprised 36.13% of all cases (81/227). Of the 126 benign lesions, the majority were inflammatory in nature (68/126), the next commonest lesion being lipoma (38/126). Rare cases of tuberculosis involving the sternum, epithelioid leiomyosarcoma, and neuroendocrine tumors involving ribs, malignant nerve sheath tumors involving the chest wall, metastatic carcinoma of the stomach and prostate, and papillary carcinoma thyroid are reported in this series. Histopathology was available in 24 cases, and hematological correlation in one case. A 100% cytohistological and cytohematological correlation was found, with no false positives or false negatives. In conclusion, fine-needle aspiration cytology is a rapid, diagnostic tool, eliciting many interesting lesions in the chest wall. It is useful not only in detecting primary and metastatic lesions, but also in follow-up of tumor recurrence. PMID- 11391819 TI - Pulmonary alveolar proteinosis: a spectrum of cytologic, histochemical, and ultrastructural findings in bronchoalveolar lavage fluid. AB - Pulmonary alveolar proteinosis (PAP) is defined as abundant extracellular proteinaceous periodic acid-Schiff (PAS)-positive material which represents surfactant distending alveolar spaces. While this lesion is defined by histologic findings, there are characteristic radiologic features and cytologic findings in bronchoalveolar lavage (BAL) specimens that together may provide a confident diagnosis. The BAL specimens from all patients for which a diagnosis of PAP was made or suggested on either cytologic or biopsy specimens at University of North Carolina Hospitals from 1990-1999 were reviewed. There were 23 cytologic specimens from 11 patients. Patient ages ranged from 6 wk to 76 yr. All 23 specimens had slides prepared for Papanicolaou stain, 22 specimens (all patients) had Diff-Quik stains, 10 specimens (6 patients) had PAS stains, and 8 specimens (5 patients) had lipid stains. Nine patients had lung biopsies in addition to cytologic specimens. The clinical charts of all patients were reviewed. Twenty one cytologic specimens were described as cloudy or milky, and 2 were bloody. By chart review and/or biopsy results, 8 patients were felt to have definite PAP. The initial lavage specimens from 6 of these patients showed classic cytologic findings of PAP, consisting of paucicellular specimens dominated by adundant extracellular granular to globular material which was basophilic on Diff-Quik stain, pale to focally eosinophilic on Pap stain, and PAS-positive, diastase resistant. Five of these patients had biopsies; 3 showed PAP, and 2 were insufficient. Later BAL specimens after therapeutic lavage from these patients were often less characteristic, with scant extracellular material present. The other 2 patients with PAP clinically and by biopsy had atypical cytologic findings, with one showing numerous macrophages with scant PAS-positive material and abundant lipid mimicking lipid pneumonia, and one showing moderate eosinophils in addition to the extracellular proteinacous material. The remaining 3 patients were felt not to have PAP clinically or by biopsy (1 lymphocytic interstitial pneumonitis, 1 rheumatoid lung, and 1 hemosiderosis), and their BAL specimens predominantly contained macrophages with rare proteinaceous extracellular globules. Electron microscopy was performed in 5 patients (4 considered to have PAP, and 1 with lymphocytic interstitial pneumonitis) and in all cases showed whorled myelin figures characteristic of surfactant. The PAP cases and the non-PAP case had identical ultrastructural findings. We conclude that BAL specimens with classic cytologic features and supporting clinical and radiographic evidence may be diagnosed as PAP. Atypical specimens should be approached with caution, and may represent either PAP or other pulmonary diseases with secondary accumulation of surfactant. Cytology specimens taken subsequent to therapeutic lavage from PAP patients may also not be diagnostic. PMID- 11391820 TI - Fine-needle aspiration of hepatocellular carcinoma: False-negative result due to epithelioid cells. AB - The presence of epithelioid cells in fine-needle aspirations of a liver nodule is rare, but may complicate the diagnosis of the nodule. We report on a case of a liver nodule in hepatitis C cirrhosis. Results of fine-needle aspiration mainly revealed the presence of epithelioid cells, without any recognizable tumor cells. Histological examination of the nodule after surgical resection showed a hepatocellular carcinoma with numerous epithelioid and gigantocellular granulomas, without necrosis. PMID- 11391821 TI - Aspiration cytology of the collagenized variant of mammary myofibroblastoma: a case report with review of the literature. AB - Myofibroblastoma of the breast is a rare benign stromal neoplasm, which occurs primarily in men. Classical myofibroblastoma is a circumscribed, nonencapsulated tumor comprised of bipolar fusiform cells arranged randomly, or in fascicles alternating with broad collagenous bands. Additional histologic variants (the cellular, collagenized, infiltrative, and epitheloid types) have been described. Several case reports describe the cytopathologic features of the classical and cellular variants. We report on a 70-yr-old woman, who presented with a circumscribed mass in her left breast. Aspiration biopsy showed paucicellular smears with singly distributed atypical spindle-shaped cells and rare fragments of collagenized stroma, raising suspicion of a phyllodes tumor. Histologic examination revealed spindle-shaped cells distributed in a diffusely collagenized stroma. Some nuclear atypia was present. To the best of our knowledge, this is the first case reporting the cytologic features of the collagenized variant of myofibroblastoma. Although we believe a specific diagnosis of myofibroblastoma can be rendered in a male based on the typical cytologic and clinical findings in the classical type, the variant forms are difficult to classify accurately and require excision for a definitive diagnosis. PMID- 11391822 TI - Myoepithelioma presenting as a midline cystic tongue lesion: cytology, histology, ancillary studies, and differential diagnosis. AB - Salivary gland myoepithelioma (ME) is a neoplasm derived from myoepithelial cells that lacks the ductal and broad mesenchymal differentiation seen in the vast majority of mixed tumors. This report describes the cytologic findings of a cystic ME presenting in the midline of the dorsal tongue, a site where no salivary glands are generally present. The tumor was well circumscribed and composed of sheets of monotonous epithelioid cells without ductal cells. The cells were positive for S-100 protein and ultrastructurally had features of myoepithelial cells. The fine needle aspiration (FNA) biopsy findings, differential diagnosis, histology, immunohistochemistry, and electron microscopic features of this interesting and uncommon neoplasm are presented. To the best of our knowledge, there have been no cytologic reports of ME of the tongue. PMID- 11391823 TI - Plasmacytoma of the larynx diagnosed by fine-needle aspiration cytology: a case report. AB - Extramedullary plasmacytoma is a rare lesion. The use of fine-needle aspiration for diagnosis of plasmacytoma has been described in a few sporadic reports. To the best of our knowledge, none of these reports described the cytologic findings from plasmacytoma of the larynx. We report on a case of laryngeal plasmacytoma in a 79-yr-old man diagnosed by fine-needle aspiration cytology. The patient had a history of a plasmacytoma involving the sixth thoracic vertebra diagnosed in 1996, which progressed to multiple myeloma in 1997. He received treatment in the form of local radiation to the skeletal vertebrae and chemotherapy. Two years later, the patient presented with a large neck mass. Computed tomography (CT) was done at an outside facility, and the radiologic impression was of a large right glottic carcinoma with invasion into the right thyroid cartilage. Because of the history of multiple myeloma, a fine-needle aspiration (FNA) biopsy was performed of the laryngeal mass. Cytologic examination demonstrated atypical plasma cells arranged in a dissociative fashion, consistent with a plasmacytoma. Although there are previous surgical pathology reports of laryngeal plasmacytoma, to the best of our knowledge, this is the first report of plasmacytoma of the larynx diagnosed by FNA cytology. PMID- 11391824 TI - Cost-effectiveness of monolayers and human papillomavirus testing compared to that of conventional Papanicolaou smears for cervical cancer screening: protocol of the study of the French Society of Clinical Cytology. AB - The French Society of Clinical Cytology is conducting a study to compare the cost effectiveness of monolayers and human papillomavirus (HPV) testing with that of conventional Papanicolaou (Pap) smears for cervical cancer screening. The protocol of this study is presented. It includes 3,000 women who will be evaluated by the three methods (conventional Pap smears, or monolayers with or without HPV testing) and by the reference method: colposcopy followed, in cases with abnormalities, by cervical biopsy. Efficacy or performance of the methods will be compared on the basis of sensitivity. Cost comparisons and cost effectiveness modeling will be based on the costs associated with methods themselves and also the costs of "false positives." This will require specific collection of data concerning the costs of the three methods, as these costs have not previously been accurately documented. Patient recruiting and data collection started in September 1999 and will be complete in June 2000. The first results are expected to be available in spring 2001. PMID- 11391825 TI - Comparison of fine-needle aspiration cytology and core biopsy for diagnosis of breast cancer. AB - Both fine-needle aspiration (FNA) cytology and core biopsy are useful in the diagnosis of breast cancer. In order to compare the sensitivities of these procedures, we reviewed 209 patients with breast cancer who had either FNA, core biopsy, or both, and also either mastectomy or lumpectomy. Sensitivities for FNA and core biopsies for diagnosing breast cancer were calculated and compared. Sensitivity for FNA or core biopsies interpreted as either atypical or malignant was 93.8% for FNA and 90.1% for core biopsy (P > 0.05). Sensitivity for FNA or core biopsies interpreted as malignant was 65.4% for FNA and 88.7% for core biopsy (P < 0.0001). Sensitivities of FNA interpreted as either atypical or malignant were 92.4% for FNA performed by pathologists and 100% for FNA by nonpathologists (P > 0.05). Sensitivities of FNA interpreted as malignant were 75.8% for FNA by pathologists and 20.0% for FNA by nonpathologists (P < 0.00001). Both FNA and core biopsies are sensitive procedures for the detection of breast cancer. There was no significant difference between sensitivity of FNA and core biopsies interpreted as either atypia or malignancy, although the sensitivity of core biopsies interpreted as unequivocal malignancy was greater than that of FNA. FNAs performed by pathologists were more sensitive than FNAs performed by nonpathologists in making an unequivocal diagnosis of breast cancer. PMID- 11391826 TI - Neural network-based screening (NNS) in cervical cytology: no need for the light microscope? AB - Neural network-based screening (NNS) of cervical smears can be performed as a so called "hybrid screening method," in which parts of the cases are additionally studied by light microscope, and it can also be used as "pure" NNS, in which the cytological diagnosis is based only on the digital images, generated by the NNS system. A random enriched sample of 985 cases, in a previous study diagnosed by hybrid NNS, was drawn to be screened by pure NNS. This study population comprised 192 women with (pre)neoplasia of the cervix, and 793 negative cases. With pure NNS, more cases were recognized as severely abnormal; with hybrid NNS, more cases were cytologically diagnosed as low-grade. For a threshold value > or = HSIL (high-grade squamous intraepithelial lesions), the areas under the receiver operating characteristic (ROC) curves (AUC) were 81% (95% CI, 75-88%) for pure NNS vs. 78% (95% CI, 75-81%) for hybrid NNS. For low-grade squamous intraepithelial lesions (LSIL), the AUC was significantly higher for hybrid NNS (81%; 95% CI, 77-85%) than for pure NNS (75%; 95% CI, 70-80%). Pure NNS provides optimized prediction of HSIL cases or negative outcome. For the detection of LSIL, light microscopy has additional value. PMID- 11391827 TI - Phagocytosis of erythrocytes by Trichomonas vaginalis: examination of a cervicovaginal smear. PMID- 11391828 TI - p27(Kip1) Protein expression in Hashimoto's thyroiditis diagnosed by fine-needle biopsy. PMID- 11391829 TI - An interview with Hugh A. Gamble, II, MD--MSMA's 134th President. PMID- 11391830 TI - Payment error prevention: 'PEPPing' up the program. PMID- 11391831 TI - Frequency of hepatitis C virus antibodies and elevated serum alanine transaminase levels in Ghanaian blood donors. AB - The objective of the study was to determine the frequency of Hepatitis C Virus (HCV) positivity in blood donors and investigate the possibility of using serum alanine transaminase (ALT) elevation as a surrogate marker. Blood samples from 1300 healthy blood donors were screened for HCV antibodies by an ELISA technique. The serum ALT was also assayed in 725 of them. 68 (5.2%) were positive for HCV antibodies. In the ALT assay 23 (3.2%) had elevated values and only 13 of this number had ALT values of 80 I.U and above. Only two positive HCV donors had elevated serum ALT. It is concluded that 5.2 per cent of blood donors had HCV antibodies and ALT assay cannot be used as a surrogate marker. PMID- 11391832 TI - Pain assessment in Nigerians--Visual Analogue Scale and Verbal Rating Scale compared. AB - The usefulness of 2 methods of pain assessment was determined in a cohort of Nigerians who had pain as a symptom and were receiving physiotherapy for various indications. The English and Yoruba versions of two Pain Rating Scales, the Visual Analogue Scale (VAS) and Verbal Rating Scale (VRS) with 1 to 4 or 1 to 5 intensity scales (VRS-4 or VRS-5) were employed for the assessment of pain in 100 patients. The mean pain score on the 4-point VRS scale was 2.49 +/- 0.72, for the 5-point VRS 2.1 +/- 1.18 and for the VAS 4.93 +/- 2.5. Correlation analysis for corresponding groups of patients showed a significant positive relationship between the VAS and VRS-4 (r = 0.68 P < 0.001) VAS and VRS-5 (r = 0.64 P < 0.001) indicating that both VAS and VRS constitute useful tools for pain assessment in Nigerian patients. PMID- 11391833 TI - Iatrogenic urological trauma: a 10-year experience from Port Harcourt. AB - A retrospective study was done to define the clinical characteristics and outcome of treatment of iatrogenic urological injuries in Port Harcourt, Nigeria. Consecutive cases of iatrogenic urological injuries treated by consultant surgeons based in the University of Port Harcourt Teaching Hospital over a period of 10 years were reviewed. A total of 37 injuries occurred in 34 patients. The ages ranged from two weeks to 74 years with a mean of 30 years. The distribution of these injuries by sex was 23 males and 14 females. The operations in which the injuries occurred were: hysterectomy 12 cases (32%), hernia repairs 8 cases (22%) and male circumcision 6 cases (16%). The organs injured were ureter 13 times (35%), the bladder 12 times (32%) and the glans penis and distal urethra 12 times (32%). The surgeons responsible were mainly as follows: Gynecologist/Obstetrician 14 (38%), General Practitioner 9 (24%), Nurses 4 (11%), Non-medical persons 4 (11%), Not disclosed 3 (8%). The outcome of treatment was satisfactory in 33 (90%). There were two deaths. Strategies to prevent these injuries include adequate surgical training and meticulous surgical techniques. PMID- 11391834 TI - Determination of metabolite and regulatory enzyme levels in Dirofilaria immitis and Ascaris suum: a comparative study. AB - Studies on metabolite levels in Dirofilaria immitis revealed similarities in several metabolites with those of Ascaris suum. The glycogen level in the filariid was however 3-4 times lower than that in A. suum. Levels of three regulatory enzymes were also determined in D. immitis and compared with those in A. suum. The activities of Hexokinase and Phosphofructokinase were similar. However, the levels of Glycogen phosphorylase b appeared to be much lower in the filariid than in A. suum. The subtle but important differences observed may reflect modifications of the parasite enzymes suggesting salient differences in the regulation of energy production from carbohydrates in the worms. The differences may also represent specialization required for the unique life style of the worms in their different locations in their hosts. PMID- 11391835 TI - Renal cell carcinoma in Enugu, Nigeria. AB - Over a period of ten years (July 1988 to June 1998) all the patients seen at two health institutions in Enugu metropolis, with histologically diagnosed renal cell carcinoma, were studied to elucidate the pattern of presentation, management and response to treatment. Seventy four (74) such patients were seen during the study period. Fifty two (70.3%) were males while 22 (29.7%) were females. The ages of the patients ranged between 17 years and 72 years with a mean of 44 years. The commonest features at presentation were weight loss (100%) raised ESR (94.6%), haematuria (86.5%), anaemia (66.2%) and upper quadrant mass (64.9%). No bilateral case was encountered. Thirty two patients (43.2%) presented with the clinical triad of haematuria, loin pain and renal mass. Seven patients (9.5%) were HIV positive. Fifty nine patients (79.7%) presented with stages 3 and 4 disease and the outcome was poor. Those that presented with stages 1 and 2 tumour had good prognosis. Early diagnosis of the tumour is important in order to give the patients a high chance of survival. PMID- 11391836 TI - Interferon alfa-2a (Roferon-A) in the management of chronic hepatitis B infection: results of an open prospective study in Nigerian patients. AB - The efficacy and safety of recombinant interferon alfa-2a (rIFN) was evaluated in 26 adult Nigerian patients with chronic hepatitis B infection. Male and female patients with serological evidence of HBV infection (HBsAg and/or HBeAg positive patients) and abnormal liver histology were monitored for six months to confirm chronicity. At the end of the six months screening period eligible patient were enrolled into the study and treated with rIFN 4.5 MIU given three times a week for 6 months. Efficacy was assessed primarily by loss of HBV-DNA and/or HBeAg from serum and secondarily by loss of HBsAg and normalization of the liver histology. Safety was assessed by monitoring the leukocyte and platelet count over the treatment period whilst tolerability was assessed by recording the occurrence of adverse events (adverse drug reaction and intercurrent illness). At the end of therapy the response rate with respect to loss of HBV-DNA was 67% and 100% for HBeAg (i.e. for the six patients who were HBeAg positive at baseline). There was loss of HBsAg in 22.2% of the patients. A significant reduction in inflammation and necrosis scores was found among the 10 patients who had both baseline and term biopsies. The frequency of occurrence of adverse events was 53.8% and the laboratory safety parameters were not significantly affected by therapy (p > 0.05). 19.2% of the enrolled patients were withdrawn from the study prematurely. These results demonstrate that rIFN is effective in the management of CHB infection even in Nigerians. The high success rate associated with HBcAg clearance is particularly noteworthy. PMID- 11391838 TI - The incidence of glycosuria in patients attending the Korle bu and Mamprobi polyclinics. AB - A survey conducted to search for the incidence of glycosuria among the patients who attended consultation at the Polyclinics at Korle bu and Mamprobi is published. Sample population of 800 patients who came to these clinics for the first time had specimen of their urine collected at random, and tested for glucose. Out of these 800 patients 633 were of general cases while 167 were pregnant women who came to attend pre-natal consultation services. Out of the general cases, there were 33 cases of glycosuria; an incidence of 4.1%. Among the pregnant women, there were 5 cases of glycosuria; an incidence of 0.63%. The highest incidence of glycosuria was observed within the age group of (21-30 years). Glycosuria among the pregnant women is more rampant among women were more than among men. The incidence among the pregnant women was not more than 0.63.0% compared with about an incidence of 3% in nonpregnant women. PMID- 11391837 TI - Management of HIV infection in Nigeria with zalcitabine in combination with saquinavir mesylate: preliminary findings. AB - The efficacy and safety of a combination therapy with two anti-retroviral drugs, zalcitabine (ddC) and saquinavir mesylate was evaluated in 24 adult Nigerian patients with HIV infection. The result of an interim analysis after a 6-month course of therapy is presented herein. Patients were given zalcitabine 2.25 mg and saquinavir 1800 mg per day. Efficacy was evaluated by improvement in the CD4 cell count and disappearance and/or resolution of clinical signs and symptoms from the patient baseline condition. Tolerability and safety were assessed by the occurrence of adverse event and monitoring of biochemical parameters such as alanine transaminase, alkaline phosphatase and total bilirubin. The haemogram profile of patients was also monitored. There was clinical improvement in 79.2% of the patients, a minimal increase in the CD4 cell count was observed and the incidence of adverse event was 40%. The haematological and biochemical profile of the patients were not significantly affected by treatment (p > 0.05). We therefore conclude that the drug cocktail comprising zalcitabine and saquinavir does posses good potentials for effective management of Nigerian patients with HIV infection. However, it is imperative and important to continue treatment with the drugs for a longer time in order to demonstrate sustained response. PMID- 11391839 TI - Colorectal adenocarcinoma in children and adolescents: a report of 8 patients from Zaria, Nigeria. AB - Colorectal adenocarcinoma is predominantly a disease of the old and less than 1% of patients are below 20 years in most reports. Though increasingly younger patients are seen in Africa, most reports indicate that the disease is extremely rare in children and adolescents. This is a report of 8 patients below 20 years managed for colorectal adenocarcinoma in a 10-year period in Zaria, Nigeria. They represented 16.3% of all cases of colorectal adenocarcinoma seen at the institution, an incidence higher than that in other parts of Africa and developed countries. All the tumours were in the rectosigmoid region and are accessible to digital rectal examination and proctosigmoidoscopy. The histology was mucinous adenocarcinomas in 6 patients, 5 of who had a Duke's stage C or D disease and well-differentiated in 2 (Duke's stage B and C respectively). Haemorrhoids was found in association in 2 patients and schistosoma mansoni in one and were responsible for delay in referral and diagnosis. Only palliative treatment could be offered in most patients due to advanced disease. Three patients died within 7 months and one after 2 years from their disease. One patient died from sepsis following surgery. The outcome in 3 patients could not be ascertained. It is emphasized that children and adolescents with rectal bleeding must have digital rectal examination and proctosigmoiscopy as this is the only hope of making an early diagnosis. PMID- 11391840 TI - Intra-operative cardiac arrest--a tropical experience. AB - In order to determine the pattern of intra-operative cardiac arrests in a developing country, we reviewed 40 consecutives cases of cardiac arrests at the Jos University Teaching Hospital between January 1993 and December 1997. During this period 15,060 minor cases and 9800 medium/major surgical procedures were performed and an arrest rate of 1:15,060 for minor cases and 1:251 for medium/major cases were obtained. The age ranged between one day and 65 years with a mean of 29.8 years and a male/female ratio of 1:1.3 as there were 18 males and 22 females. Twenty-five (62.5%) arrests occurred outside work hours while fifteen (37.5%) cases arrested during work hours. The surgical procedures with high arrest rates in this study were: emergency laparotomy 8 (20%), emergency caesarian sections 7 (17.5%), thoracotomy 6(15%), emergency craniotomy 5(12.5%), emergency hysterectomy 4(10%) and therapeutic bronchoscopy for foreign bodies in the airway 4(10%). The arrests occurred in 30(75%) emergency procedures as opposed to 10(25%) elective cases. Only in 3 out of the 18, 318 day case procedures did the patients arrest. One patient arrested during local infiltration of lignocaine while the other 39(97.5%) arrested under general anaesthesia. There was no arrest with spinal anaesthesia. The predisposing factors for a patient to arrest on the operating table in our environment include emergency major surgery, poor risk patients with ASA 111 and above, surgery performed outside work hours, under general anaesthesia administered by nurse anaesthetists or junior anaesthetic residents. The success rate at resuscitation is highest with patients with ASA 1 & 11, operations performed during work hours and by senior surgeons and anaesthetists. PMID- 11391841 TI - An inexpensive and available external fixator. AB - This article introduces a simple and inexpensive external fixator device which has been designed and manufactured from materials readily available in this country. It is called the Doxa Fixator. PMID- 11391842 TI - Diet, alcohol consumption, smoking and exercise as determinants of blood lipid levels of Nigerians. AB - Diet, alcohol consumption, smoking and exercise habits of randomly chosen 250 Nigerians were investigated by questionnaires. Most of the subjects, 67% were on carbohydrate diets, 50% were teetotal, 87.5% were non-smokers and 69% were not involved in exercise. The mean blood lipid levels of the Nigerians were within normal range, but much lower than the lipid levels of caucasians. It was concluded that the lifestyle of these Nigerians was largely responsible for the low levels of plasma lipids observed. PMID- 11391843 TI - Interferon alfa-2a (Roferon-A) monotherapy in chronic myelogenous leukemia: a pilot study in Nigerian patients in early chronic phase. AB - The efficacy and safety of interferon alfa-2a monotherapy was evaluated in seventeen Nigeria patients with chronic myelogenous leukaemia (CML). Male and female patients with a mean age of 34.5 +/- 10.6 years were recruited into the study. Interferon therapy was administered at a maintenance dose of 9 MIU daily for 12 months. Efficacy was evaluated by assessing both haematologic and cytogenetic response, tolerability by incidence of adverse events and safety by laboratory haematological and biochemical indices. At the end of 12 months of therapy 6 patients (54.4%) had complete haematologic remission whilst 3 patients (100% of those evaluated) showed partial cytogenetic remission. The incidence of adverse event was 70% and the monitored haematologic and biochemical indices were not adversely affected by treatment. In conclusion, the study clearly demonstrated a significant benefit of interferon alpha-2a in the management of Nigerian patients with CML. The changes in the haematological and cytogenetic profiles between baseline and term were significant (p < 0.05). However, it is imperative and important to encourage and continue monitoring of the responding and stabilized patients beyond 12 months in order to demonstrate sustained response. The drug was reasonably well tolerated, however life threatening pancytopenia may pose a major problem in certain cases. PMID- 11391844 TI - Treatment of malaria in north-eastern and south-eastern Nigeria: a population study of mefloquine, sulphadoxine, pyrimethamine combination (MSP) vs chloroquine (CQ). AB - In a population-based study involving 4019 patients in 20 peripheral health facilities in Nigeria, the outcome of presumptive malaria treatment with MSP was compared to that of CQ. The study was conducted between January 1995 and January 1996. Patients aged 6 months or more with a clinical diagnosis of malaria based on history of fever and axillary temperature > 37.5 degrees C were either treated with MSP (250 mg mefloquine, 500 mg sulphadoxine, and 25 mg pyrimethamine per tablet) or CQ (150 mg chloroquine base per tablet). The clinical cure rate was assessed by the disappearance of clinical signs and symptoms over a 7-day period. Tolerability was assessed by the incidence of adverse events (adverse drug reaction and intercurrent illness). The result shows that the clinical care rate of suspected malaria was 97.6% with MSP and 85.6% with CQ. The incidence of adverse event was 9.5% with MSP and 9.2% with CQ. The withdrawal rate was 2.0% with MSP and 5.0% with CQ; 3.5% of the patients in the CQ group withdrew due to adverse events compared to 0.47% with MSP. In conclusion it was observed that in addition to superior efficacy of MSP over CQ, fever clearance rate with MSP was comparable to that of CQ. The study also demonstrated that two tablets maximum dose of MSP is safe and effective in a large population of Nigeria malaria patients. PMID- 11391845 TI - An audit of prescribing practices in health care facilities of the Wassa West district of Ghana. AB - While many health districts recognise that irrational prescribing leads to wastage of drugs, few monitor prescribing practices. We investigated drug use in government health facilities in the Wassa West district of Ghana. Retrospective prescribing data were obtained from 700 outpatients' clinical record cards in 7 government health facilities in the district. Prescribing patterns were significantly worse in the health centres than in the district hospital in terms of polypharmacy, use of injectable drugs and antibiotics. Overall, 4.8 drugs were prescribed per patient, 97% of drugs were on the national essential drugs list and 65% of drugs were prescribed by their generic names. Antibiotics and injectable drugs were prescribed for 60% and 80% patients respectively. The observed patterns were related to the cadre of prescribers, availability of diagnostic facilities, participation in recent refresher training and patient demand. Measures to improve prescribing practices are discussed. PMID- 11391846 TI - Pattern and outcome of medical admissions at the Ogun State University Teaching Hospital, Sagamu--a three year review. AB - This is a three-year retrospective study of the pattern and outcome of acute medical admissions at the Ogun State University Teaching Hospital, Sagamu. Our findings showed that there were 1,938 admissions over the study period of which 1,044 (54%) were females and 894 (46%) were males. The ages of the patients ranged between 12 to 86 years with a mean of 49 +/- 1.7 years. The age range was 14 to 80 years for males with a mean of 47 +/- 6.1 years and 12 to 86 years for females with a mean of 49 +/- 4.5 years. The length of stay was between 5 to 25 days with a mean of 15 +/- 0.5 days. There were 16 patients per bed per year with a turn around of 8 days and bed occupancy of 65% 194 (10%) patients discharged themselves against medical advice and there were 488 (25%) deaths. The interval between admission and death ranged between 4 to 7 days with a mean of 5.5 +/- 0.07 days. Indications for admissions were infectious diseases (38%), neurological disorders (19.6%) gastro-intestinal disorders (11%), genito-urinary tract disorders (10.2%) endocrine disorder (10%), cardiovascular disorders (9.9%), respiratory disorders (6.4%) and haematological disorders (4.9%). Tuberculosis accounted for 10% of total medical admissions and was the commonest disease entity responsible for medical admissions. Acquired immuno deficiency syndrome (AIDS) constituted 1.8% of medical admissions. Mortality was highest for infections accounting for 32% of deaths while malaria accounted for the lowest. The reasons for the relative frequency and mortality are adduced. The study recommends improvement of preventive strategies towards communicable diseases in the community and encourages better admission policy, provision of appropriate facilities and manpower to improve the hospital services. PMID- 11391847 TI - Herd immunity to measles virus in a population of student nurses and medical students following a widespread outbreak of clinical measles in Ibadan, Nigeria. AB - Following an outbreak of serologically--and virologically--confirmed measles requiring large-scale hospitalisation of children in Ibadan, Nigeria, the herd immunity to measles virus among medical students and student nurses was determined. Of the 200 students tested, none lacked haemagglutination--inhibiting antibody to measles virus. The titre of HI--antibody ranged from 2(5) to 2(10). Describing a titre of 2(9) as very high, a significantly higher proportion of student nurses than medical students (P < 0.05) had very high antibody titres to measles virus. There was however no statistical difference between the sexes (P > 0.05). Using a commercial Enzyme Immuno Assay kit (EIA), anti measles IgM could not be detected from any of the students. Thus a clear evidence of recent infection with measles virus during the outbreak could not be detected among the students, a probable indication that student nurses and medical students may not participate in the maintenance of wild measles virus within the hospital environment in developing countries like Nigeria. PMID- 11391848 TI - Extremity gunshot injuries in civilian practice: the National Orthopaedic Hospital Igbobi experience. AB - A combined retrospective and prospective study of Gunshot Injuries (GSI) that presented to the National Orthopaedic Hospital, Igbobi (NOHL) between 1991 and 1995 was undertaken with the aim of determining the characteristics of these injuries in Lagos, Nigeria. 232 patients with 281 gunshot wounds and 212 gunshot fractures were seen during this period. 68.9% of patients in the study were in the age group 21-40 years with a mean age at presentation of 32.46 +/- 11.21 years. The male to female ratio was 9:1. 87 (37.5%) presented within 6 hours of injury. Armed robbery dominated the events surrounding the shootings with high velocity weapon (HVW) accounting for 47% of the cases. While the femur was the commonest single bone to be fractured the treatment of fractures generally was largely conservative as only 5 fractures were eventually treated by open reduction and internal fixation. Wound infection was the commonest complication (25%) with amputation being performed in 5.6% of cases. This study does not confirm the belief that high velocity weapon causes greater morbidity than low velocity weapon. Even though the average duration of hospitalisation was 33.5 +/- 23.4 days, we advise that for our present state of development gunshot fractures should not be primarily treated with internal fixation. PMID- 11391849 TI - A case report: malignant haemangiopericytoma in infancy. AB - A case of malignant haemangiopericytoma of the right foot in an infant is reported. Nodules were noticed on the foot at birth and these gradually increased in size to involve the whole foot on presentation at the age of ten months. With a working diagnosis of haemangioma, tumour excision and foot reconstruction was done. After histological diagnosis, ablation was offered but refused. PMID- 11391851 TI - Preliminary study of fortnightly irinotecan hydrochloride plus cisplatin therapy in patients with advanced gastric and colorectal cancer. AB - PURPOSE: Irinotecan hydrochloride shows a strong activity against gastric cancer and colorectal cancer, while combined therapy with irinotecan and cisplatin is useful for gastric cancer. However, myelosuppression and diarrhea are still dose limiting factors. To reduce such toxicities to enable therapy to be performed on an outpatient basis, we tested the effect of divided administration of cisplatin. METHODS: Irinotecan (60 mg/m2) plus cisplatin (30 mg/m2) were administered on days 1 and 15 every 4 weeks to 13 patients with advanced gastric cancer and 13 with advanced colorectal cancer. Treatment was continued if a leukocyte count > or = 3000/mm3, a platelet count > or = 100,000/mm3, and grade 0 diarrhea were confirmed. Doses were reduced if grade 3-4 hematological toxicity and grade 2 or higher nonhematological toxicity occurred. RESULTS: The major toxicity was leukopenia (neutropenia), but grade 3-4 nonhematological toxicity was not observed. The response rate was 41.7% for gastric cancer (5/12 evaluable patients) and 36.7% for colorectal cancer (4/11 evaluable patients). The median survival time was 313 days (range 29-920 days) for gastric cancer patients and 490 days (range 83-1184 + days) for colorectal cancer patients. CONCLUSION: Fortnightly administration of irinotecan and cisplatin (with a divided cisplatin dose) seems to be a useful regimen for gastrointestinal cancer. It reduces toxicity while maintaining a good antitumor effect. PMID- 11391850 TI - Measured versus estimated glomerular filtration rate in the Calvert equation: influence on carboplatin dosing. AB - PURPOSE: Carboplatin is frequently dosed to achieve a desired area under the plasma concentration-time curve (AUC) by using the Calvert or Chatelut equations to estimate carboplatin clearance. Accurate determination of glomerular filtration rate (GFR) is necessary to correctly calculate carboplatin clearance using the Calvert equation. In clinical practice, the Cockcroft-Gault formula is frequently used to estimate GFR, but this practice has been reported to under- and overestimate carboplatin clearance. The purpose of this trial was to compare determinations of carboplatin clearance using the Chatelut equation and four separate GFR determinations, including 99mTc-DTPA, the Cockcroft-Gault formula, a 24-h urine collection and a 2-h urine collection. METHODS: Carboplatin clearance was estimated in 21 previously untreated extensive-stage small-cell lung cancer patients. GFR was determined using 99mTc-DTPA, the Cockcroft-Gault formula, 24-h urine collection and 2-h urine collection. Serum and urine creatinine concentrations were measured using enzymatic assays. The carboplatin clearance was then calculated by individually adding 25 to the four GFR determinations based on the Calvert equation, which states that carboplatin clearance equals GFR + 25 (nonrenal clearance). The carboplatin clearance was also estimated using the Chatelut equation. The five determinations of carboplatin clearance were compared using Friedman's test and post-hoc Wilcoxon signed rank tests. Precision and bias for each carboplatin clearance determination were calculated assuming that 99mTc DTPA provided the most accurate measure of GFR. RESULTS: A statistically significant difference was found between the five methods of estimating carboplatin clearance (P < 0.001). No difference was found between carboplatin clearance calculated using 99mTc-DTPA and the Chatelut equation, the Cockcroft Gault formula or the 2-h urine collection. The Chatelut equation provided more precision and less bias than the 2-h urine collection (median precision 20% and 30%, median bias -1% and -18%, respectively). CONCLUSION: Compared to 99mTc-DTPA, the Chatelut equation more accurately estimates carboplatin clearance than the Cockcroft-Gault formula, the 2-h urine collection and the 24-h urine collection. The greater negative bias found for the latter three estimates of carboplatin clearance could result in underdosing of carboplatin. PMID- 11391852 TI - Pharmacokinetics and cerebrospinal fluid penetration of phenylacetate and phenylbutyrate in the nonhuman primate. AB - INTRODUCTION: Phenylbutyrate (PB) and its metabolite phenylacetate (PA) demonstrate anticancer activity in vitro through promotion of cell differentiation, induction of apoptosis through the p21 pathway, inhibition of histone deacetylase, and in the case of PB, direct cytotoxicity. We studied the pharmacokinetics, metabolism, and cerebrospinal fluid (CSF) penetration of PA and PB after intravenous (i.v.) administration in the nonhuman primate. METHODS: Three animals received 85 mg/kg PA and 130 mg/kg PB as a 30-min infusion. Blood and CSF samples were obtained at 15, 30, 35, 45, 60 or 75 min, and at 1.5, 2.5, 3.5, 5.5, 6.5, 8.5, 10.5 and 24.5 h after the start of the infusion. Plasma was separated immediately, and plasma and CSF were frozen until HPLC analysis was performed. RESULTS: After i.v. PA administration, the plasma area under the concentration-time curve (AUC) of PA (median +/- SD) was 82 +/- 16 mg/ml.min, the CSF AUC was 24 +/- 7 mg/ml.min, clearance (Cl) was 1 +/- 0.3 ml/min per kg, and the AUCCSF:AUCplasma ratio was 28 +/- 19%. After i.v. PB administration, the plasma PB AUC was 19 +/- 3 mg/ml.min, the CSF PB AUC was 8 +/- 11 mg/ml.min, the PB Cl was 7 +/- 1 ml/min per kg, and the AUCCSF:AUCplasma ratio was 41 +/- 47%. The PA plasma AUC after i.v. PB administration was 50 +/- 9 mg/ml.min, the CSF AUC was 31 +/- 24 mg/ml.min, and the AUCCSF:AUCplasma ratio was 53 +/- 46%. CONCLUSIONS: These data indicate that PA and PB penetrate well into the CSF after i.v. administration. There may be an advantage to administration of PB over PA, since the administration of PB results in significant exposure to both active compounds. Clinical trials to evaluate the activity of PA and PB in pediatric central nervous system tumors are in progress. PMID- 11391853 TI - Paclitaxel administration on days 1 and 8 every 21 days in anthracycline pretreated metastatic breast cancer patients. A multicenter phase II trial. AB - Paclitaxel is now included in second- and even first-line regimens in advanced breast cancer. The optimal dose and schedule of this drug, however, still remain a matter of investigation. A group of 57 consecutive patients with advanced breast cancer previously treated with anthracycline-containing regimens were submitted to treatment with single-agent paclitaxel administered at 130 mg/m2 on days 1 and 8 every 21 days. Of the 57 patients, 56 were fully evaluable, and of these 25 had an absolute anthracycline resistance, 14 a relative resistance and 17 were potentially sensitive. The median age of the patients was 57 years (range 33-71 years), their median performance status was 1 (0-3), and 27 (47%) had liver involvement, 17 (30%) lung involvement, 30 (53%) bone involvement and 15 (26%) skin/lymph node involvement. Toxicity was recorded in 295 cycles. This scheme was well tolerated, the dose-limiting toxicities being hematological and neurological. Grade 3/4 leukopenia was observed in 20% of patients at nadir, while grade 3 leukopenia was observed in 3% of patients at recycle. Only one patient experienced febrile neutropenia. Grade 2/3 neurotoxicity was observed in 26% of patients, leading to drug withdrawal in three. The treatment was given on an outpatient basis in all patients and the median relative dose intensity of 86.6 mg/m2 per week was 100% of the planned dose (range 75-100%). Three patients (5%) attained a complete clinical response and 12 (21%) a partial response for an overall response rate of 26% (95% confidence interval 18-38%), while 30 (53%) attained disease stabilization and 11 progressed (19%). Time to progression in responding patients was 10.3 months, and the median overall survival of the entire population was 15.4 months. To conclude, paclitaxel administration on days 1 and 8 every 21 days was active and manageable in advanced breast cancer patients previously treated with anthracyclines. The response obtained was durable. PMID- 11391854 TI - Epidermal growth factor enhances cisplatin-induced apoptosis by a caspase 3 independent pathway. AB - PURPOSE: Activation of the epidermal growth factor (EGF) receptor has previously been shown to increase the sensitivity of cancer cells to DNA-damaging agents, including cisplatin, UV-B, and gamma-radiation. We now investigated the mechanisms by which EGF enhances the sensitivity of human ovarian cancer cells to cisplatin. RESULTS: The effect of EGF on cisplatin sensitivity could not be entirely explained by alterations in the cellular detoxification of cisplatin by glutathione or DNA repair of transcribed genes, as assessed by a plasmid reactivation assay. Furthermore, EGF did not affect the levels of several proteins that regulate apoptotic pathways, including bcl2, bclXL, bax and p53. Cisplatin treatment resulted in activation of caspase 3 and subsequent cleavage of specific substrates containing the DEVD (Asp-Glu-Val-Asp) amino acid sequence, including PARP (poly(ADP-ribose) polymerase). The EGF-mediated increase in cisplatin-induced apoptosis, however, did not result in increased caspase 3 activity. Moreover, apoptosis induced by cisplatin alone was completely inhibited by the caspase 3 inhibitor DEVD-CHO, whereas cell death induced by combined treatment with cisplatin and EGF was not prevented by inhibition of caspase 3. CONCLUSION: Our results suggest that, although cisplatin alone induces apoptosis by a caspase 3 dependent pathway, EGF enhances cisplatin-induced cell death by activating an apoptotic pathway that is independent of caspase 3. PMID- 11391855 TI - Population pharmacokinetic modeling and model validation of a spicamycin derivative, KRN5500, in phase 1 study. AB - PURPOSE: KRN5500, a novel spicamycin derivative, shows the greatest activity against a human tumor xenograft model and the highest therapeutic index among spicamycin derivatives. KRN5500 is currently under clinical development in Japan and the United States. The objective of this study was to develop a population pharmacokinetic model that describes the KRN5500 plasma concentration versus time data. METHODS: Data were collected from 18 patients entered in a phase 1 study. These patients received KRN5500 3-21 mg/m2 as a 2-h infusion. A total of 219 concentration measurements were available. The data were analyzed using the nonlinear mixed effect model (NONMEM) program. In addition, the basic and final population pharmacokinetic models were evaluated using bootstrapping resampling. RESULTS: The basic model selected was a two-compartment model with a combination of additive and constant coefficient of variation error models. The basic model fitted well not only the original data, but also 100 bootstrap replicates generated from the original data set. With regard to the effect of covariates selected by generalized additive modeling analysis, gender (SEX) and performance status were found to be possible determinants of the volume of central compartment by NONMEM analysis. The final regression model for V1 was V1 = theta V1 (1--SEX x theta SEX), where V1 is the typical population value of the volume of central compartment, and SEX = 0 if the patient is male, otherwise SEX = 1. The final model was fitted to the 200 bootstrapped samples. The mean parameter estimates were within 15% of those obtained with the original data set. CONCLUSIONS: The KRN5500 plasma concentration versus time data obtained from the phase 1 study were well described by the population pharmacokinetic model. Further evaluation by bootstrapping showed that the population pharmacokinetic model was stable. PMID- 11391856 TI - Plasma and cerebrospinal fluid pharmacokinetics of gemcitabine after intravenous administration in nonhuman primates. AB - PURPOSE: Gemcitabine (dFdC) is a difluorine-substituted deoxycytidine analogue that has demonstrated antitumor activity against both leukemias and solid tumors. Pharmacokinetic studies of gemcitabine have been performed in both adults and children but to date there have been no detailed studies of its penetration into cerebrospinal fluid (CSF). The current study was performed in nonhuman primates to determine the plasma and CSF pharmacokinetics of gemcitabine and its inactive metabolite, difluorodeoxyuridine (dFdU) following i.v. administration. METHODS: Gemcitabine, 200 mg/kg, was administered i.v. over 45 min to four nonhuman primates. Serial plasma and CSF samples were obtained prior to, during, and after completion of the infusion for determination of gemcitabine and dFdU concentrations. Gemcitabine and dFdU concentrations were measured using high performance liquid chromatography (HPLC) and modeled with model-dependent and model-independent methods. RESULTS: Plasma elimination was rapid with a mean t1/2 of 8 +/- 4 min (mean +/- SD) for gemcitabine and 83 +/- 8 min for dFdU. Gemcitabine total body clearance (ClTB) was 177 +/- 40 ml/min per kg and the Vdss was 5.5 +/- 1.0 l/kg. The maximum concentrations (Cmax) and areas under the time concentration curves (AUC) for gemcitabine and dFdU in plasma were 194 +/- 64 microM and 63.8 +/- 14.6 microM.h, and 783 +/- 99 microM and 1725 +/- 186 microM.h, respectively. The peak CSF concentrations of gemcitabine and dFdU were 2.5 +/- 1.4 microM and 32 +/- 41 microM, respectively. The mean CSF:plasma ratio was 6.7% for gemcitabine and 23.8% for dFdU. CONCLUSIONS: There is only modest penetration of gemcitabine into the CSF after i.v. administration. The relatively low CSF exposure to gemcitabine after i.v. administration suggests that systemic administration of this agent is not optimal for the treatment of overt leptomeningeal disease. However, the clinical spectrum of antitumor activity and lack of neurotoxicity after systemic administration of gemcitabine make this agent an excellent candidate for further studies to assess the safety and feasibility of intrathecal administration. PMID- 11391857 TI - Heterogeneic distribution of thymidine phosphorylase between primary tumors and metastatic lesions of human pancreatic ductal carcinoma: implications for the efficacy of chemotherapy with 5-FU or its derivatives. AB - PURPOSE: It has been suggested that the expression of thymidine phosphorylase (TdRPase) correlates with the malignant potential of various cancers, but its involvement in human invasive ductal carcinoma (IDC) of the pancreas has not been reported. In the present study, the distribution and clinical significance of TdRPase in IDCs and benign diseases of the pancreas were assessed, especially in relation to the efficacy of chemotherapy with 5-FU or its derivatives. METHOD: The expression of TdRPase in 148 specimens of pancreatic IDCs (66 primary lesions, 46 nodal lesions and 36 distant metastases from 126 patients) and in 24 specimens of benign diseases (4 cystadenomas, 3 hyperplasias, and 17 chronic pancreatitises) was examined by immunohistochemical staining with anti-TdRPase monoclonal antibody and evaluated in terms of three grades of immunoreactivity: negative 0, low 1, or high 2. RESULTS: Positive TdRPase staining (low and high immunoreactivity) was detected in 71% (47/66) of the primary lesions, in 46% (21/46) of the involved nodes, in 53% (19/36) of various lesions of distant metastasis, and in 37% (9/24) of the benign diseases. The staining intensity was significantly higher in the IDC tissues than in the benign disease tissues, and significantly lower in the metastatic lesions than in the primary lesions. TdRPase reactivity did not correlate with the survival rate in both resectable and unresectable IDCs. In patients with both primary tumor and nodal involvement, however, high TdRPase activity in involved nodes was significantly associated with a poor prognosis. On the other hand, although adjuvant chemotherapy was found to improve the survival of patients, TdRPase activity in the tumor did not show any significant relationship with the efficacy of chemotherapy with 5-FU or its derivatives. CONCLUSIONS: The present study suggested that in pancreatic IDC the activity of TdRPase in primary lesions is different from that in metastatic lesions, and that DNA is synthesized mainly through the salvage pathway in primary lesions and through a de novo pathway in metastatic lesions. This may be one of the reasons for the heterogeneity in chemosensitivity of human pancreatic IDC. PMID- 11391858 TI - Sequential intrahepatic and systemic fluoropyrimidine-based chemotherapy for metastatic colorectal cancer confined to the liver. A phase II study. AB - PURPOSE: Several trials have suggested that intrahepatic chemotherapy increases the likelihood of response in advanced colon cancer patients, but has no significant impact on survival due to the development of extrahepatic metastases. We report our experience of combined hepatic intraarterial and systemic chemotherapy in advanced colorectal cancer patients. METHODS: A group of 35 patients received intrahepatic FUDR (0.3 mg/kg per day for 14 days by continuous infusion), followed, after 1 week's rest, by systemic 5-FU and L-leucovorin (370 and 100 mg/m2 per day, respectively, both for 5 consecutive days). After another week off therapy, the combined intrahepatic and systemic regimen was repeated and cycles continued until disease progression. RESULTS: Of 32 assessable patients, 17 (53.1%) had an objective response, while 8 (25%) had disease stabilization. Median time to progression (TTP) was 32 weeks (range 8-104 weeks), while the median overall survival was only 39 weeks (range 9-109 weeks). Incomplete liver perfusion was the only variable that showed a significant correlation with a poorer survival (P = 0.046, log-rank test). CONCLUSIONS: Our results are in agreement with previous data suggesting a relative efficacy of such a treatment approach for advanced colon cancer patients. More thorough investigations are warranted, especially as an adjuvant treatment for selected high-risk patients. PMID- 11391859 TI - The vitamin D3 analog, ILX-23-7553, enhances the response to adriamycin and irradiation in MCF-7 breast tumor cells. AB - Ionizing radiation and the anthracycline antibiotic, Adriamycin, generally fail to promote a primary apoptotic response in experimental breast tumor cell lines. Similarly, the primary response of breast tumor cells to vitamin D3 (1,25(OH)2D3) and vitamin D3 analogs such as EB 1089 is growth inhibition. Previous studies have demonstrated that pretreatment of MCF-7 breast tumor cells with vitamin D3 or EB 1089 can increase sensitivity to both Adriamycin and irradiation. PURPOSE: The capacity of the vitamin D3 analog, ILX 23-7553, to enhance the antiproliferative and cytotoxic effects of Adriamycin or irradiation and to promote apoptosis in MCF-7 breast tumor cells was assessed in the present study. RESULTS: Pretreatment of MCF-7 cells with ILX 23-7553 followed by Adriamycin or irradiation decreased viable cell numbers by 97% and 93%, respectively. Cell numbers were reduced by 56%, 74% and 75% by ILX 23-7553, Adriamycin and irradiation alone. Pretreatment with ILX 23-7553 also shifted the dose response curve for clonogenic survival, increasing sensitivity to Adriamycin 2.5-fold and sensitivity to radiation fourfold. In addition, ILX 23-7553 pretreatment conferred sensitivity to Adriamycin- or irradiation-induced DNA fragmentation and resulted in morphological changes indicative of apoptotic cell death in MCF-7 cells. Statistical analysis demonstrated that ILX 23-7553 interacts additively and not synergistically with both Adriamycin and irradiation. CONCLUSIONS: ILX 23 7553 enhances the effects of Adriamycin and irradiation in MCF-7 breast tumor cells by decreasing viable cell numbers, reducing clonogenic survival and inducing apoptotic cell death. Current studies are focused on elucidating the mechanisms underlying the induction of apoptosis as well as understanding the nature of the interactions between ILX 23-7553 and Adriamycin or irradiation. PMID- 11391860 TI - Antagonism of oestrogen action in human breast and endometrial cells in vitro: potential novel antitumour agents. AB - PURPOSE: There is a need to find novel oestrogen receptor (ER) ligands that antagonize oestrogen action in the reproductive tissues and would therefore have therapeutic potential in oestrogen-dependent tumours. We tested novel ER ligands in both breast and endometrial cells to profile agonism/antagonism in these oestrogen target reproductive tissues. METHODS: Novel analogues of the ER antagonist ICI 182,780 were synthesized and tested for their ability to inhibit gene expression dependent on oestrogen response elements (ERE) in human breast (MCF-7) and endometrial (Ishikawa) cell lines. This activity was correlated with inhibition of oestrogen-induced cell proliferation and ER binding. RESULTS: The sulphide analogue (compound 1) and sulphone analogue (compound 2) had no intrinsic ERE-dependent agonism in either breast cancer or endometrial cells in culture. All three compounds dose-dependently inhibited ERE-mediated oestrogen agonism. Moreover, these ER ligands inhibited oestrogen-stimulated proliferation of breast cancer and endometrial cells. ICI 182,780, compound 1 and compound 2 were all able to bind both isoforms of the ER (ER alpha and ER beta). In endometrial cells, the relative binding to ER beta correlated with the ERE dependent antioestrogenic effect of these ligands, suggesting that in this tissue this receptor is the predominant isoform that determines antioestrogenic activity. CONCLUSIONS: The ability of these analogues of ICI 182,780 to inhibit oestrogen-stimulated transcriptional activity and cell proliferation suggests that these agents, in particular the sulphone analogue, have therapeutic potential in the treatment of breast cancer without exhibiting the unwanted oestrogenic effects in the endometrium. PMID- 11391861 TI - Taxol-induced ceramide generation and apoptosis in human breast cancer cells. AB - PURPOSE: Taxol has emerged as a valuable antimitotic chemotherapeutic agent, particularly in advanced breast and ovarian cancers. Although much is known about cytotoxic mechanisms, the effectiveness of Taxol cannot be solely explained by microtubular interaction. This study was undertaken to determine whether ceramide generation plays a role in Taxol-induced apoptosis. METHODS: Hormone-independent MDA-MB-468 and hormone-dependent MCF-7 breast cancer cell lines were employed, and ceramide metabolism was characterized using [3H]palmitic acid as lipid precursor. RESULTS: Exposure of cells to Taxol resulted in enhanced formation of [3H]ceramide. Ceramide increased nearly 2-fold in MDA-MB-468 cells exposed to 50 nM Taxol, and more than 2.5-fold in MCF-7 cells exposed to 1.0 microM Taxol. These concentrations mirrored the EC50 (amount of drug eliciting 50% cell kill) for Taxol in the two cell lines. Use of cell-permeable C6-ceramide as a medium supplement revealed that MDA-MB-468 cells were 20-fold more sensitive to ceramide than MCF-7 cells (P < 0.001). Ceramide was generated as early as 6 h after exposure to Taxol in MDA-MB-468 cells, whereas the earliest signs of apoptosis were detected 12 h after treatment, and by 24 h the apoptotic index was six times that of untreated cells. Both fumonisin B1, a ceramide synthase inhibitor, and L cycloserine, a serine palmitoyltransferase inhibitor, blocked Taxol-induced ceramide generation, whilst sphingomyelin levels remained unchanged, indicating a de novo pathway of ceramide formation. L-Cycloserine reduced Taxol-induced apoptosis by 30% in MDA-MB-468 cells and totally blocked Taxol-induced apoptosis in MCF-7 cells. CONCLUSIONS: These results suggest that Taxol-induced apoptosis is, in part, attributable to ceramide and sphingoid bases. This is of relevance to drug mechanism studies, as ceramide is a known messenger of apoptosis. Clinical use of Taxol with ceramide-enhancing agents may maximize cytotoxic potential. PMID- 11391862 TI - Improved respiratory delivery of the anticancer drugs, camptothecin and paclitaxel, with 5% CO2-enriched air: pharmacokinetic studies. AB - PURPOSE: To increase pulmonary deposition of anticancer liposome aerosols in mice by modulation of respiratory physiology through the addition of 5% CO2 to the air source used to generate the aerosols. Breathing CO2-enriched aerosol increases pulmonary ventilation with concurrent increased deposition of inhaled particles. METHODS: Dilauroylphosphatidylcholine liposome formulations of two anticancer drugs, paclitaxel (PTX) and camptothecin (CPT), were investigated. The aerosol droplet size was measured using an Andersen cascade impactor. Drug concentrations in aerosol droplet fractions and tissues were determined by HPLC analysis. ICR mice were exposed to each liposome aerosol for 30 min. For each drug, one group of mice inhaled the drug-liposome aerosol generated with a mixture of 5% CO2 in air and another group inhaled the drug-liposome aerosols produced with normal air. Tissue distribution and pharmacokinetics were determined for both drug delivery systems. RESULTS: Significantly higher concentrations of PTX and CPT were found in organs of mice exposed to 5% CO2-air aerosols compared to organs of mice exposed to normal air aerosols. The highest concentrations of drug were detected in the lungs and were two- to fourfold higher with 5% CO2-air aerosols than with aerosols generated with normal air. Higher concentrations were also detected in liver, spleen, kidneys, blood, and brain. CONCLUSION: 5% CO2 enrichment of air increased respiratory tract deposition of inhaled aerosol particles containing PTX and CPT. PMID- 11391863 TI - A pharmacological study of the weekday-on/weekend-off oral UFT schedule in colorectal cancer patients. AB - PURPOSE: The new weekday-on/weekend-off schedule for oral UFT administration consists of its administration for 5 consecutive days followed by 2 days off the drug. The intratumor 5-FU (5-fluorouracil) concentration has been reported to be correlated to the tumor response in patients treated with intravenous 5-FU. The aim of this study was to investigate the pharmacokinetics during the 2 days off the drug in cancer patients treated with the weekday-on/weekend-off schedule for oral UFT. METHODS: The subjects were 24 colorectal cancer patients. They were divided into three groups, and were all given UFT, 600 mg/day, for 5 days before surgery. Surgery was performed 2, 24, or 48 h after the final dose of UFT. The 5 FU concentrations in the serum, tumor, and in the normal mucosa were measured. RESULTS: The serum 5-FU concentrations after the final dose of UFT were: 23 +/- 12 ng/ml (mean +/- SD) at 2 h, 7 +/- 3 ng/ml at 24 h, and 6 +/- 3 ng/ml at 48 h. The intratumor 5-FU concentrations were: 113 +/- 45 ng/g at 2 h, 54 +/- 20 ng/ml at 24 h, and 54 +/- 35 ng/ml at 48 h, and the concentrations in the normal mucosa were: 36 +/- 15 ng/g (mean +/- SD) at 2 h, 17 +/- 6 ng/ml at 24 h, and 18 +/- 6 ng/ml at 48 h after the final dose. While the serum 5-FU concentration decreased to very low levels by 24 h after the final dose of UFT, the intratumor 5-FU concentrations were maintained at more than 50 ng/g at least until 48 h after the final dose. The 5-FU concentrations in the normal mucosa were maintained at about one third of the intratumor concentrations at all time points. CONCLUSION: Although the weekday-on/weekend-off schedule for UFT administration included intermittent 2-day drug-off periods, this pharmacokinetic study revealed that the 5-FU concentrations in the tumor were maintained at much higher levels than in the serum throughout these periods. PMID- 11391864 TI - Special issue commemorating the thirtieth anniversary of Eiji Osawa's C60 paper. PMID- 11391865 TI - How the news that we were not the first to conceive of soccer ball C60 got to us. PMID- 11391866 TI - Fullerene science--a most international endeavor. PMID- 11391867 TI - Simulation of geometrical and electronic structure of quasi-two-dimensional layer consisting of fullerenes D6h-C36. AB - This article describes a computer simulation of the geometrical and electronic structure of a quasi-two-dimensional carbon layer with a trigonal lattice consisting of fullerenes C36 (1) with topological symmetry D6h. Every polyhedral cluster 1 of this polymeric layer (2) is surrounded by six similar fullerenes and connected with every such a fullerene by two covalent bonds. Atomic coordinates of the repeating unit are estimated on the basis of MNDO/PM3 calculations of hydrocarbon molecule (D6h)-C132H48 (3). The carbon skeleton of 3 coincides with a sufficiently large fragment of the polymeric layer 2. The electronic spectrum of the quasi-two-dimensional layer 2 is calculated by the crystalline orbital method in the EHT approximation. The band gap in the electronic spectrum of 2 was found to be equal to 1.5 eV. The geometric and electronic structure of some oligomers of cluster C36, quasi-linear macromolecule [C36]n, and "hypergraphite" layer is also discussed. PMID- 11391868 TI - Weighted HOMO-LUMO energy separations of properly closed-shell fullerene isomers. AB - The HOMO-LUMO energy separation has been used as an index of kinetic stability for fullerenes. However, its utility is rather limited, in that a larger fullerene molecule in general has a smaller HOMO-LUMO energy separation even if it is kinetically stable. The T value, i.e., the HOMO-LUMO energy separation weighted with the number of carbon atoms, is free from this kind of difficulty. Properly closed-shell isomers of fullerenes with antibonding LUMOs were found to have an exceptionally large T value irrespective of the number of constituent carbon atoms and are hence predicted to be kinetically very stable. Thus, the antibonding character of the LUMOs is one of the determinants for kinetically stabilizing fullerene molecules. PMID- 11391869 TI - Addition patterns, codes and contact graphs for fullerene derivatives. AB - The mathematical concept of the d-code and its associated contact graph give a model for sterically constrained addition patterns in fullerene derivatives C60Xm and C70Xm. In combination with simple electronic arguments, the stoichiometries, symmetries, and location of addends can be predicted, yielding a small number of candidates for further study. For example, sterically and optimal solutions C60Xm with pairwise separation of d bonds between addends are found at m(d) = 24(2), 12(3), 6(4,5), 2(6 to 9). The solution for C60X24 is unique, and the model selects 12 candidates for C60X12 from a starting set of 11661527060 possibilities. PMID- 11391870 TI - Efficient generation of the cartesian coordinates of truncated icosahedron and related polyhedra. AB - Efficient algorithms for deriving the analytical expressions of the rectangular coordinates of the vertices of regular polyhedra and truncated icosahedron inscribed in a cube is described and the results are exposed. Various characteristic quantities of the geometrical structure of truncated icosahedron are obtained. Kaleidoscopes for projecting the truncated icosahedron are discussed. PMID- 11391871 TI - Planar rearrangements of fullerenes. AB - A notion of planar rearrangement of fullerenes is proposed as a general framework for all conceivable rearrangements of fullerenes satisfying a minimum physicochemical condition. The planar rearrangement is defined as an edge relocation such that the planarity of a given Schlegel diagram remains undisturbed. Graph-theoretical properties of planar rearrangements are discussed and characteristics for establishing a hierarchy between them are pointed out. A number of graph-theoretical concepts have been introduced to provide means for classification and systematic generation of fullerene rearrangements. The simpliest nontrivial rearrangements is shown to be the Stone-Wales rearrangement. PMID- 11391872 TI - Computing the relative gas-phase populations of C60 and C70: beyond the traditional delta H(fo),298 scale. AB - Computations and experiments have shown that the relative heat of formation (i.e., the heat of formation per carbon atom) of C70 is lower than of C60. Moreover, various computations suggest that this is actually a general trend among fullerene cages. The relationship is particularly important for gas-phase fullerenes. Experiments have shown that C60 is typically more populated than C70 when produced in high-temperature gas-phase synthesis. It is not immediately obvious how to reconcile those two terms, or whether the relative heats of formation and the relative populations are in conflict or in agreement. This article deals with this problem, treating it as a general task of relative stabilities of gas-phase clusters of different dimensions (i.e., nonisomeric clusters) under different types of thermodynamic equilibria. The results are then applied to C60 and C70 and point out that the conventional standard pressure of 1 atm is considerably different from actual fullerene-synthesis conditions. Apparently, we should expect considerably lower cluster pressures in carbon-arc synthesis. At 1 atm, C70 is more populated than C60, but at the conditions of a saturated carbon vapor the stability order is reversed in favor of C60 so that an agreement with experiment is obtained already within the thermodynamic treatment. The pressure effects are modeled using the MNDO, AM1, PM3, and SAM1 quantum chemical semi-empirical methods as well as the available experimental data. The computations consistently show that, if the pressure effects are considered, C60 becomes more populated than C70. Relationships of the thermodynamic treatment to more sophisticated but impractical kinetic analysis are also discussed. PMID- 11391873 TI - Curvature matching and strain relief in bucky-tori: usage of sp3-hybridization and nonhexagonal rings. AB - The relief of different types of curvature strain in bucky-tori of elemental carbon is considered in a general formal framework. This theory then is used to aid in the design of several structures, which are treated via molecular mechanics. Novel illustrations of the remnant strain are made, and some modest conclusions as to the nature of the structure of the experimentally observed bucky-tori are suggested. PMID- 11391874 TI - The molecular structure and electronic spectrum of the C@C60 endohedral complex: an ab initio study. AB - We performed ab initio quantum-chemical studies at the HF and B3LYP levels of theory to determine the geometry and electronic spectra of the C@C60 complex. The STO-3G and 6-31G(d) basis sets were employed. Two different types of stable conformations for the endohedral atom related to the centers of five- and six member rings were found. The estimated potential barrier between those conformations is small; under certain conditions, the endohedral atom can transfer from one location in the molecular cage to another. The influence of the location of endohedral carbon atom on the electronic spectrum of C60 is discussed. PMID- 11391875 TI - A sightseeing tour in the world of clusters--serendipity and scientific progress. AB - The discovery of the fullerenes in 1985 by Kroto et al. and the development of a method for producing macroscopic amounts in 1990 by Kratschmer et al. opened a new area of carbon research and the possibility of producing new materials with unique properties. The field has developed further with discoveries of nanotubes, metal-filled nanotubes, carbon onions, met cars, and metal-covered fullerenes, all of which have unique properties and possible technical applications. In retrospect, it is in interesting that indications of many unique species have existed before their discovery but have not been recognized. In general, it seems that one is so focused on a given problem that one does not realize how many great discoveries or serendipities are "hidden" in available experimental and theoretical data. In addition to generating a lot of scientific progress, these new discoveries in the field of cluster science, and in particular in carbon species such as fullerenes and nanotubes, have opened up the doors to different areas of science such as mesoscopic physics and modern material science. The general trend is from small to large systems, contrary to the general trend of modern mesoscopic physics or microelectronics where the movement is from large to small. It is especially fascinating how the whole area of fullerene research was initiated to solve problems in astrophysics. Originally, Kratschmer and Huffman intended to explain an observed strong extinction from interstellar dust; in experiments they produced a special carbon soot with a characteristic optical absorption known as "camel hump smoke." Furthermore, the original interest of Kroto was also to solve problems in astrophysics, while Osawa in his original paper on calculations of carbon molecules in organic chemistry focused on how different bonding of carbon atoms would give special species. He then found the truncated icosahedral built up of sixty carbons, without noticing its unique structure, which is today the famous C60. PMID- 11391876 TI - Intrafullerene electron transfers in Sm-containing metallofullerenes: Sm@C2n (74 < or = 2n < or = 84). AB - The electronic properties of Sm-containing metallofullerenes, Sm@C74, Sm@C76 (I, II), Sm@C78, Sm@C80, Sm@C82 (I, II, III) and Sm@C84 (I, II, III), are characterized by UV-Vis-NIR absorption spectroscopy and electron energy-loss spectroscopy (EELS). The UV-Vis-NIR absorption spectra of Sm@C74, Sm@C80, Sm@C82 (I, II, III) and Sm@C84 (I, II) are quite similar to those of the corresponding Ca, Sr, Ba, Eu, Tm, Yb-based metallofullerenes. In contrast, the absorption spectra of Sm@C76 (I, II), Sm@C78 and Sm@C84(III) show a novel feature: the onset for Sm@C78 is observed approximately 2600 nm, which corresponds to a small band gap (approximately 0.5 eV). Furthermore, the oxidation states of Sm atom in the various fullerene cages are investigated by EELS, which reveals that the Sm atom takes +2 oxidation state in the fullerene cages. A probable rationale for the tendency to have the Sm2+ state is presented based on a simple thermochemical cycle model. PMID- 11391877 TI - Mg@C72 MNDO/d evaluation of the isomeric composition. AB - Temperature development of the relative stabilities of isomers of Mg@C72 (which has not yet been isolated) is computed using the recently introduced MNDO/d method. Four isomers originally considered for the Ca@C72 case are treated: one isolated-pentagon-rule (IPR) structure, two structures with a pair of adjacent pentagons, and one cage with a heptagon. The IPR structure comes as the lowest in MNDO/d potential energy, being rather closely followed by the two structures with a pentagon-pentagon pair. On the other hand, the structure with a heptagon is located too high in potential energy to be of any experimental significance. The entropy contributions are evaluated by the MNDO/d-based partition functions so that the relative concentrations can be treated accordingly. The computations suggest that if Mg@C72 is isolated, it should be a mixture of either two or three isomers. The prediction depends on temperature prehistory. If preparation takes place at temperatures of approximately 1000 K, two isomers should be produced. If temperatures are increased to approximately 2000 K, there will already be three isomers with significant relative concentrations. The study supplies a further interesting example of the profound role of enthalpy-entropy interplay in stabilities of isomeric fullerenic structures. PMID- 11391878 TI - A theoretical study of transition metal complexes of C60 and C70 and their ring opened alternatives. AB - Ring opened structures of C60 and C70 are shown to be stabilized by complexation with transition metal fragments of the form CnHnM, where n = 3 to 6 and M = Cr, Mn, Fe, Co, and Rh. The ring opening of C60 and C70 is compared with the reverse process of the well-known catalytic conversion of acetylene into benzene. Calculations at the semi-empirical PM3(tm) level show that the 6-membered ring in C60 and C70 can be opened up in different ways through complexation with transition metal fragment. The mode of ring opening depends on the number of external 5- and 6-membered rings around the 6-membered ring being cleaved. The structures and energetics of the various ring-opened structures are discussed. PMID- 11391879 TI - Tailoring the curvature, bowl rigidity and stability of heterobuckybowls: theoretical design of synthetic strategies towards heterosumanenes. AB - Quantum mechanical calculations predict that larger heteroatom substituents on the periphery increase the feasibility of the crucial third ring closure in sumanene and are responsible for the accompanying modulations in the curvature, rigidity, stability and some of the physicochemical properties of the resulting heterosumanenes. Systematic application of semiempirical, ab initio, and DFT methods reveal that the qualitative trends obtained and our principal conclusions are independent of level of theory, albeit with minor quantitative differences. PMID- 11391880 TI - Why the all-electron full-potential approach is suitable for calculations on fullerenes and nanotubes? AB - Already 30 years have passed since the first prediction of C60 by Professor Osawa. A family of cage-type fullerenes and carbon nanotubes were experimentally found as the third form of carbon molecules in the 1980s. After this discovery, much research has been conducted experimentally and theoretically on these new materials. The all-electron full-potential approach is important for fully understanding the quantum mechanical behavior of the fullerenes and related molecules. We show some results of band calculations and ab initio molecular dynamics. PMID- 11391881 TI - Using e-mail to facilitate nursing scholarship. AB - A central tenet of university education is the development of scholarly abilities. The traditional hallmarks of scholarship, such as critical thinking, reflection, creativity, critical analysis, and openness to new thinking, need to be fostered in the face of increasing technological change. Contemporary e-mail applications provide a sophisticated environment in which many of the fundamental skills of scholarship can be nurtured. This article describes an interdisciplinary curricular project in which e-mail was employed as a vehicle to support the development of students' understanding of scholarship within a cohort of first-year undergraduate nursing students. The project involved the establishment of "Virtual Colleague" activity, which used e-mail to conduct scholarly activities, including critical analysis of electronic journals and web sites and peer review. The theoretical underpinning of the project and an evaluation of the project are presented. Future use of such technologies in the promotion of scholarship among nurses is discussed. PMID- 11391882 TI - Knowledge acquisition and evaluation of an expert system for managing disorders of the outer eye. AB - The aim of the current study was to develop and evaluate an expert system for diagnosing outer eye disorders, intended to support family physicians and nurse practitioners in the management of patients with minor eye complaints. The knowledge in the program was gathered from the scientific literature and from 6 experienced ophthalmologists and 6 ophthalmic nurses. Fourteen diagnoses must be considered, and the differential diagnosis is dependent upon 32 signs and symptoms. The program calculates the most likely diagnosis, presents photographs of the typical ocular findings, and provides information regarding management and treatment. The program's output was compared with the diagnosis made by 8 other ophthalmologists at a Swedish university clinic, based on data from 157 patients with disorders of the outer eye, who visited the emergency ward during a one-week period. The concordance between the diagnosis made by the ophthalmologists and the program was 96%. In conclusion, the expert system for disorders of the outer eye provided essentially the same diagnostic information as an examination by an ophthalmologist. Support from the program would make it possible for family physicians and nurse practitioners to manage most of these patients. PMID- 11391883 TI - Survey and workshop initiative on community nurses' knowledge of the Internet. AB - Information technology in the nursing profession has been in existence for years, but only now with the growing presence of the Internet is it viewed as a possible platform to help meet the need for nurses' continuing educational development. For this to happen, it is important to know qualified nurses' experience with the Internet. In the United Kingdom, there is little understanding of whether community nurses have experience in using the Internet and if they do, how they use it. This article reports the findings of a survey questionnaire, taken in 1998, that was designed to target a random sample of 100 community nurses in Aberdeen, Scotland, to gauge awareness and attitudes toward the use of electronic sources of information for continuing education purposes. The response rate for the questionnaire was 83% (n = 83). A series of workshops was also undertaken, exploring the potential of the Internet and its access. A total of 52 people attended the workshops. One of the main aims of the workshops was to demystify the image of the computer, breaking down barriers for those with little or no experience in this field. The major findings of the survey and workshops indicate that although many respondents were interested in the potential benefits of the Internet to access information, there was little use made of and limited access to computer equipment. PMID- 11391884 TI - Full implementation of a computerized nursing records system at Kochi Medical School Hospital in Japan. AB - A project to fully implement a novel computerized nursing records system resulted in the standardization of nursing records, reduced the administrative workload for staff, enabled medical staff to know a patient's status at any given time, and improved the quality of nursing care provided to patients. The development process of the computerized nursing records system involved three main steps: 1) the establishment of a new nursing assessment form and introduction of nursing diagnosis into routine work, 2) computerized system design and construction, and 3) the usability check of the computerized nursing records system in a clinical setting for 1 year. The successful development of the computerized nursing records system was based on the following points: 1) the assessment form for nursing diagnosis was improved and the nursing diagnosis was introduced before the computerized system was designed and constructed; 2) full, rather than partial, implementation of the computerized system occurred; 3) existing knowledge of nursing assessments and standard care planning were fully used; 4) registered data were optimally reused upon summarization and readmission to reduce the nurses' workload; and 5) portable computers were introduced to enable simple and quick recording of bedside findings. The routine use of the computerized nursing records system was started in April 2000. More comprehensive investigations during the next 2 to 3 years are necessary to determine how the contents of nursing records can be improved and how much the computerized nursing records system affects the quality of nursing. PMID- 11391885 TI - Predicting the learning and consultation time in a computerized primary healthcare clinic. AB - Managers would like to know how long it takes healthcare service providers to achieve the same throughput of patients per day that they were used to with a pen and-paper system. This study has been undertaken to derive a model for predicting the time it takes a service provider from a previously disadvantaged community to enter a patient's record in terms of his or her experience and the number of data units that have to be captured. A model was also derived to predict the average consultation time in terms of the number of data units that are captured by an experienced service provider. It can be inferred that healthcare service providers should be allowed at least 6 months of computerized system experience before any decisions about the success of the technology introduction can be made. PMID- 11391886 TI - The complexity of developing a nursing information system: a Brazilian experience. AB - Information systems development is a complex process involving technical, organizational, social, and environmental issues. The active participation of nurses in information systems development is important but not easily accomplished, especially in settings in which the nurses are not familiar with the technology. Participatory design, which focuses on community participation, is an approach to information systems development that is not often used. This article explains this approach and describes the experience of using the participatory-design approach in designing a nursing information system project in a Brazilian teaching hospital. Traditional systems- development approaches are not sensitive to individuals' needs or political and organizational dimensions. The focus of the article is on the basic topics discussed with nurses and the relevance of those topics to the design of nursing information systems. A nursing classification system is a basic component of an information system and a relevant instrument for the creation of formalized nursing knowledge. However, we should not neglect other approaches that can contribute to a better understanding of nursing work situations and the development of nursing knowledge. PMID- 11391887 TI - [Genetic variation of pine in natural Crimean populations and in artificial populations in Krivbass]. AB - The comparative analysis of the genetic variation in the plantations of Pinus pallasiana D. Don in Krivbass and the Crimean populations has been conducted using the electrophoretic division of isoenzymes of 10 geneenzymous systems, encoded by 23 loci. In the artificial plantations of industrial ecotopes of Krivbass the level of genetic variation has been higher than in natural stands of Pinus pallasiana D. Don. within the range of one microslope in the Mountainous Crimea. The Nei' s genetic distance (Dn) between two regions varied within 0.016 to 0.026. PMID- 11391888 TI - [T-DNA induced anomalies of flowers and male sterility in transgenic tobacco plants: morphometric and cytological analysis]. AB - Morphometric and cytological analysis have been carried out in transgenic tobacco (Nicotiana tabacum L.) plants possessing male sterility and abnormalities of flowers. Mutation affected flower structure and conducted to long pistil and male sterility. It was established that development of microspores was blockaded on different stages of gametogenesis but the first injuries were detected for tetrad stage. PMID- 11391889 TI - [Improvement of a series of agriculturally valuable traits by pollen selection in Linum usitatissimum L]. AB - Influence of selection of F1 high temperature tolerant pollen on the structure of BC1 segregating populations was studied for a number of agronomically important traits in oil flax. It was found that heat treatment of heterogeneous pollen population considerably changed genetic composition of plant population not only for tolerance to drought but for duration of seedling emergence--flowering period and quantity and inclination angle of side shoots as well. It is supposed that genes which confer these traits are at least partially linked to the genes responsible for pollen sensitivity to high temperature. PMID- 11391890 TI - [Analysis of quantitative traits connected with seed weight and flowering terms in birdsfoot trefoil (Lotus corniculatus L.) plants]. AB - The nature of inheritance of characters connected with the seed size (seed mass) and quality of the shoots (germination energy, seed viability, general shoot length, shoot mass, dry weight of shoots) in Lotus corniculatus L. was analyzed. The investigations were carried out on plants of three varieties: the wild form from the Krasnodar Region and local forms MF1 and MF3. The correlation analysis was carried out. The pattern of inheritance in the terms of plant flowering and length of shoots were studied. PMID- 11391891 TI - [Prospects for using multimarker dihaploid lines for mapping the barley genome]. AB - Model population of spring barley Oregon Wolfe Pack dihaploid lines, derived from F1 of cross between dominant marker stock and recessive line, was studied on presence of morphological markers. For detection of DNA variability RAPD-analysis of these genotypes was carried out. Polymorphic fragment correlating with morphological trait was detected using primer P89. PMID- 11391892 TI - [Participation of structural gene markers and anonymous DNA sequences in genetic differentiation in the species Ovis aries L. and Ovis nivicola borealis]. AB - Genetic variability and genetic differentiation in two Ovis species--domesticated (Askanian sheep, Sokilska and Kulunda sheep) and wild (bighorn sheep) were analyzed using different types of molecular-genetic markers--genetic-biochemical (30 loci) and DNA (ISSR-PCR) ones. High level of genetic variability was revealed. The average heterozygosity of biochemical markers loci was in range of 0.073-0.188. Markers involved into process of genofond divergence of two closely related species were revealed. PMID- 11391893 TI - [Analysis of mutations in the phenylalanine hydroxylase gene in Ukrainian families at high risk for phenylketonuria]. AB - The data on analysis of phenylalanine hydroxilase (PAH) gene mutations in 39 phenylketonuria (PKU) families from Ukraine are presented. Obtained results indicate that the most common mutation observed in the Ukrainian population is R408W mutation (66.6%). Besides two minor mutations R158Q (2.6%) and Y414C (1.25%) were revealed. PMID- 11391894 TI - [The ABO blood group system as genetic markers of quantitative traits in man]. AB - A model of correlative variability of AB0 blood groups and a quantitative trait (mad-model) was analysed. Statistics for evaluation of additive and non-additive effects of alleles IA, IB and i on quantitative trait were developed. Restrictions of the model application are discussed. The obtained results may be used in genetic epidemiology for study of sensitivity or resistance to different diseases. PMID- 11391895 TI - [Inheritance of frost resistance and winter hardiness in distant hybrids of wheat and amphiploids]. AB - The data on inheritance of frost resistance and winter hardiness of bread wheat lines obtained as a result of distant hybridization of wheat-rye and wheat-elymus amphiploids with durum and bread wheat were presented. It was shown that selection of the donors of valuable traits is sensible to make in later progenies of hybrids (F6-F7). So, it is possible to obtain the new initial breeding material for winter bread wheat selection with high frost tolerance, winter hardiness and early maturity. Hypotheses explaining the high frost resistance of hybrids are presented. The crosses of the octoploid amphiploids with durum wheat are more preferable for the alien traits introgression into wheat than the crosses with bread wheat. PMID- 11391896 TI - Do you see what I see? Illegible handwriting can cause patient injuries. PMID- 11391897 TI - Handwriting errors: harmful, wasteful and preventable. PMID- 11391898 TI - Adult and infant vocalization: speaking valves used in individuals with tracheostomy and ventilator dependency. PMID- 11391900 TI - [Chronic headache: progress in diagnosis and therapy]. PMID- 11391899 TI - A problem that carries real weight. PMID- 11391901 TI - [Chronic headache: classification and epidemiology]. PMID- 11391902 TI - [Chronic headache: the current status and future of its treatment]. PMID- 11391903 TI - [Migraine: diagnostic standards and its clinical application]. PMID- 11391904 TI - [Migraine: physiopathology]. PMID- 11391905 TI - [Serotonin and migraine]. PMID- 11391906 TI - [Migraine: selection of therapeutic agents to be applied during its attack]. PMID- 11391907 TI - [Migraine: sumatriptan]. PMID- 11391908 TI - [Migraine: use of preventive agents]. PMID- 11391909 TI - [Cluster headache: diagnostic standard and its clinical application]. PMID- 11391910 TI - [Cluster headache: clinical treatment]. PMID- 11391911 TI - [Tension headache: standards and practice of diagnosis]. PMID- 11391912 TI - [Tension headache: clinical treatment]. PMID- 11391913 TI - [Tension headache: chronic daily headache]. PMID- 11391914 TI - [Current topics: molecular biology of migraine]. PMID- 11391915 TI - [Current topics: expectation for new triptans]. PMID- 11391916 TI - [Treatment of headache, especially chronic headache: a discussion]. PMID- 11391917 TI - [MEN2A incidentally detected by high level of CEA and family history]. PMID- 11391918 TI - [Two cases of adenoid cystic carcinoma with stridor as chief complaint successfully treated with radiotherapy alone]. PMID- 11391919 TI - [Central diabetes insipidus caused by neurosarcoidosis]. PMID- 11391920 TI - [Dermatomyositis complicated with heart and lung disease and died of rapidly progressive respiratory failure]. PMID- 11391921 TI - [Hemophagocytic syndrome in a patient with systemic lupus erythematosus]. PMID- 11391922 TI - [Liver fibrosis--the forefront of the study on stellate cells]. PMID- 11391923 TI - [Leptin and cardiovascular lesions]. PMID- 11391924 TI - [Physiopathology of the development and molecular mechanism related to radiation pneumonitis]. PMID- 11391925 TI - [Standardization in hematology]. AB - The International Council for Standardization in Hematology(ICSH) is a non governmental and non-profit body devoting to standardization work on hematology tests. It has publicized over 100 standardization documents on the subjects in the past 30 years. The ICSH is currently extending its activities to cooperative work with WHO, CEN, NCCLS, ICTH, ISBT, IFCC and ISLH. ICSH-Asia has also been founded to discuss local issues on standardization in hematology among Asian countries. PMID- 11391926 TI - [Hematopoietic stem cell transplantation]. AB - Hematopoietic stem cell transplantation, which started as an experimental medicine more than four decades ago, has now become an indispensable and powerful tool to treat intractable hematologic disorders through the considerable efforts of D. Thomas and his colleagues. A short review of hematopoietic stem cell transplantation is reported with emphasis on cord blood transplantation and non myeloablative stem cell transplantation. PMID- 11391927 TI - [Cytokine]. AB - It remains to be elucidate whether measurements of cytokines in serum have any clinical significance. In this article, the clinical uses of cytokines are reviewed. The purposes of clinical use of cytokines are: 1) for the immunological treatment of malignancy; and 2) as hematopoietic growth factors. During the clinical use of cytokine, measurements of cytokine in serum might be important to estimate clinical efficacy and the clinical course. PMID- 11391928 TI - [Proceedings of the workshop "The future aspect of clinical hematology laboratory and the role of medical technologists"]. AB - In the workshop of the 34th in-service training course for University Hospital Medical Technologists in 2000 sponsored by the Ministry of Education, all participants discussed future aspects of the clinical hematology laboratory and the role of medical technologists. We report here a summary of the discussion. PMID- 11391929 TI - [Presentation of a case of hematological malignancy in reversed C.P.C]. AB - We report a 68-year old male who was admitted to our hospital with the chief complaint of fatigue. Laboratory examinations revealed: 1) macrocytic anemia; 2) leukopenia; 3) reticulocytosis; 4) reduction in serum folate level; and 5) erythroid hyperplasia in bone marrow. One year later, the patient was admitted again to our hospital. At that time, laboratory data showed: 1) leukocytosis; 2) appearance of blast cells in peripheral blood; 3) normocytic anemia; 4) thrombocytopenia; and 5) predominant proliferation of blast cell(91%) in bone marrow. Blasts showed prominent nucleoli, markedly basophilic cytoplasma with vacuolation and some blebs on cell surface. The cells were negative for peroxidase stain but positive for PAS stain. We describe the evaluation of hematological laboratory data and the diagnosis in this patient. PMID- 11391930 TI - [Reversed C.P.C. :Case of 14-year old girl with large abnormal cells in her bone marrow]. AB - A 14-year-old female was admitted to our institute with arthralgia of the left pelvis. Magnetic resonance image analysis revealed the presence of a low-density area in her left pelvis. Although the complete blood count was almost within normal ranges, large abnormal cells with a round-shaped multi-nucleus and abundant cytoplasm containing coarse basophilic granules were noted in her bone marrow smear. She was diagnosed with mastocytosis after laboratory examinations. Nine months later, after intensive chemotherapy, the patient died. PMID- 11391931 TI - [Bone marrow aspiration: practical aspects]. AB - Bone marrow aspiration is the most valuable diagnostic examination for the evaluation of hematopoiesis of the bone marrow. Appropriate smear-sampling and reliable stainings in hematological sections are essential for the precise diagnosis of bone marrow cells, and give important insight into pathological and clinical hematology. PMID- 11391932 TI - [Reversed C.P.C.: Aggressive NK-cell leukemia/lymphoma]. AB - While natural killer(NK)-cell type granular lymphocyte-proliferative disorders(GLPD) generally represent benign disorders, acute NK-cell leukemia/lymphoma is a rare but a very aggressive disorder. We report a case characterized by an extranodal(nasal) and bone marrow infiltration of large granular lymphocyte morphology, CD3-CD16-CD56+ phenotype. The patient's course became severe and refractory. Multiorgan failure, associated with intravascular coagulation syndrome was the main cause of death. PMID- 11391933 TI - [Flow cytometry]. AB - Recently, flow-cytometric analysis has become the most popular type of cell surface marker examination and it is particularly indispensable in diagnosing hematological disorders such as leukemia. Since diagnosis becomes more reliable using monoclonal antibodies, cell-surface marker analysis contributes greatly to the diagnosis of hematological disorders. In particular in lymphocytic leukemias, immunological classification becomes possible and since this classification has a close relation to pathophysiology and prognosis, it is very useful in diagnosing and treating patients with lymphocytic leukemia. Attempts at quantitative analysis of cell-surface antigens and expected benefits such as improved detail are also described. Additionally the standardization of cell-surface analysis using flow-cytometry is discussed. PMID- 11391934 TI - [FISH, cytogenetic analysis]. AB - We describe here the clinical applications of fluorescence in situ hybridization(FISH) in patients with chronic myelogenous leukemia under treatment with interferon-alpha, some cases with a false positive caused by preparation of specimen and/or probes, and the detection of chimerism after sex-mismatched bone marrow transplantation based on our clinical experience. Furthermore, we introduced our methods of performing FISH using blood smear or G banding specimens. PMID- 11391935 TI - [DNA diagnosis in hematological malignancies]. AB - The recent advances in molecular biology and gene engineering have greatly contributed to the diagnosis and treatment of hematopoietic malignancies, such as leukemia and malignant lymphoma. It is now possible to precisely determine the clonal origin of malignant cells, the subtype of leukemia or lymphoma, and the clinical prognosis in each patient. Furthermore, minimal residual malignant cells in leukemia or lymphoma patients after achieving complete remission can be detected by DNA analysis. Based on these analyses, theoretically treatment can be tailored for each patient. We discuss in the present paper the usefulness of DNA or gene analyses in the clinical laboratory for hematopoietic malignancies. PMID- 11391936 TI - [New classification of leukemia]. AB - By keeping a cytomorphological basis for the diagnosis of leukemia according to the FAB classification, a new classification system was proposed by the WHO group, which incorporated four genetically established entities of acute leukemia. This classification also made some changes in MDS categories and provided new criteria for diagnosing acute myeloid leukemia. Combined use of this classification with the FAB classification will promote investigations into leukemia. PMID- 11391937 TI - [Classification of malignant lymphomas]. AB - Hodgkin's disease is characterized by malignant tumor in lymphoid tissue, excluding non-Hodgkin's lymphomas and lymphoid leukemias. It can be diagnosed by the characteristic histologic picture, which contains Reed-Sternberg cells. The Rye classification for Hodgkin's disease has been useful to date, however, since non-Hodgkin's lymphomas were first recognized as a distinct group a variety of classifications have been proposed although none are entirely satisfactory. At present, it is concluded that the most practical approach to lymphoma categorization is simply to define the diseases with the currently available morphologic, immunologic, and genetic techniques, and use the REAL and WHO classifications for lymphoid malignancies. It is important to develop more concise and practical classifications in the future. PMID- 11391938 TI - [New laboratory tests for detecting platelet abnormalities]. AB - Platelets play important roles in thrombosis and hemostasis, and their abnormalities lead to hemostatic disorders or thrombotic diseases. The development of new tests for assessing platelet disorders is awaited. In this article, the measurement of blood thrombopoietin levels and detection of platelet aggregates with a particle counting method using light scattering are described as candidates for new laboratory tests for detecting platelet abnormalities. PMID- 11391939 TI - [Conference of morphological analysis]. AB - To date, hematological malignancies have been diagnosed and classified by morphological analysis following ordinary and special staining such as Wright's and Giemsa's double staining. The FAB classification, which is applied for acute leukemia, is a typical classification system used worldwide. Recently, various reference techniques have been developed for the diagnosis of hematological malignancies and are useful for diagnosing hematological diseases, but morphological analysis still remains the basic reference technique. Therefore, expertise in morphological analysis is extremely important, and full training in this field is necessary. PMID- 11391940 TI - [Molecular markers for the blood coagulation and fibrinolysis]. AB - It is now possible to more precisely evaluate coagulation and fibrinolysis by laboratory tests in which various molecular markers can be measured. The determination of the plasma level of soluble fibrin may be the most important molecular marker for the diagnosis of thrombosis and disseminated intravascular coagulation. Combined with the results of the other molecular marker determinations, the measurement of the plasma level of soluble fibrin might provide useful information about the coagulation status of patients. PMID- 11391941 TI - [Laboratory tests for detection of antiphospholipid antibodies]. AB - Antiphospholipid antibodies(APA) have been reported to be a heterogeneous family of immunoglobulins. Some APA can be detected via phospholipid dependent coagulation assays when they present as an aspecific coagulation inhibitor, lupus anticoagulant(LA), other antibodies can be measured via immunological assays mostly via their capability to bind to immobilized cardiolipin (anticardiolipin antibody; aCL). Despite their name, APA associated with antiphospholipid syndrome(APS) do not bind phospholipids, but are directed at plasma proteins bound with anionic phospholipids. The antigenic targets of these antibodies include beta 2 glycoprotein I(beta 2 GP I), prothrombin, high- and low-molecular weight kininogens, annexin V, protein C and protein S. In this article, the current knowledge on the methods of detection and their functional properties are reviewed, and the interaction of these antibodies and acquired activated protein C resistance are discussed. PMID- 11391942 TI - [Review of thyroid disease and recent progress in thyroid research]. AB - Major thyroid diseases and recent progress in thyroid research are reviewed, including our clinical experiences and data on genetic analysis. Of the 19,944 patients receiving care in our endocrinology and metabolism department over the past 26 years(from 1974 to 2000), there were 4,471(22.4%) patients with thyroid diseases. Of these patients with thyroid disease, 37.3% had Graves' disease, 24.1% had Hashimoto's thyroiditis, and 22.2% had a benign thyroid tumor. Male-to female ratio for Graves' disease was 1:3.2. The precise mechanism and genetic or environmental factors underlying the onset and progression of autoimmune thyroid disease need further investigation, although recent thyroid research, especially molecular level studies, has resulted in many new insights. Our genetic analysis of patients and experimental animals with thyroglobulin(Tg) abnormalities indicated the amino acids involved in the surface electric charge were important in maintaining the solid structure of Tg and thyroid hormone synthesis in addition to tyrosine and cysteine. In three patients with hyperthyroid Graves' disease, Hashimoto's thyroiditis or idiopathic hypothyroidism, followed by the author for 8 to 20 years, it was indicated that continued comprehensive care was needed for various episodes, even those arising from non-endocrine conditions, throughout the clinical course, although clinical and laboratory findings showed improvement of the thyroid disease itself. PMID- 11391943 TI - [Thyroid function tests]. AB - About 80% of thyroid disease consists of thyroid-specific autoimmune diseases, Hashimoto's disease and Grave's disease. To diagnose thyroid diseases, testings for (1) thyroid function and (2) pathogenetic autoantibodies are indispensable. To assess thyroid function, serum hormone concentrations, such as TSH, FT4 and FT3 are measured. Among these hormones, serum TSH concentrations are the most reliable and informative regarding thyroid function, correcting indicating a hyperthyroid, euthyroid or hypothyroid state. Therefore, TSH measurement appears to be the first choice in selecting the hormone determination. Reference intervals for normal healthy subjects of TSH are around 0.4-5.0 microU/ml. The second choice for thyroid function assessment are FT4 which supersedes total T4(TT4). TT4 is affected by changes in serum thyroid hormone binding proteins(TBG, TTR, Albumin). For example, euthyroid pregnant women whose serum TBG are physiologically higher than those of non-pregnant women show augmentation of TT4. However, FT4 depicts within reference intervals, although measurement of FT4 alone is unable to detect any abnormality of thyroid hormone binding proteins. According to its plasma concentration and binding affinity, FT3 measurement deserves no more significance than T3. Another important test for thyroid diseases is to detect serum autoantibodies against thyroid tissues, such as TgAb, TPOAb. Much more important is TSH receptor antibody which differentiates Graves' disease from Hashimoto's thyroiditis. In patients who show hyperthyroidism and some very uncommon hypothyroidism, TSH receptor antibodies should be measured. Three indicators are available as routine tests; TRAb measured by radioreceptor assay; TSAb determined by bioassay using cultured porcine thyroid cells. Usually, TRAb activity clinically correlates well with TSAb. TSBAb was initially discovered in patients with severe hypothyroidism with atrophic thyroid gland. TSBAb blocks thyroid stimulating activity of TSH and consequently causes severe hypothyroidism. TRAb and TSAb are very useful to diagnose and follow patients with Grave's disease. PMID- 11391944 TI - [Cardiovascular ultrasound: applications for the assessment of cardiac function]. AB - Congestive heart failure(CHF) is usually associated with impaired left ventricular(LV) systolic function, and thus, the measurement of systolic function is an essential component of the evaluation of any patients with known or suspected cardiac disease. Among many parameters, most frequently used are LV percent fractional shortening and ejection fraction(EF), which can be easily measured from an M-mode echocardiogram. However, these M-mode measurements may be inaccurate in patients with asymmetrical LV due to myocardial infarction, right ventricular overload or sigmoid septum. Especially in such cases, EF should be measured using two-dimensional echocardiography. Usually, LV volumes and EF are calculated using the disc-summation method through the manual tracing of apical two-chamber and four-chamber echocardiograms. On the other hand, it has been recognized that congestive heart failure may arise in the absence of any systolic dysfunction and CHF due to systolic dysfunction never occurs in the absence of concomitant diastolic dysfunction. Although the analysis of pulsed-Doppler transmitral flow velocity has been most widely used for the noninvasive assessment of LV diastolic function, an increase in left atrial pressure during CHF can pseudonormalize an abnormal flow pattern and mask LV diastolic dysfunction. Recently, we proposed a new index for assessing LV diastolic function, flow propagation velocity, which can be measured with color M-mode Doppler echocardiography and baseline-shift technique. Recent studies have shown that the flow propagation velocity is a unique noninvasive parameter of LV diastolic function which can accurately detect the diastolic impairment in patients with different types of cardiac diseases with various loading conditions. PMID- 11391945 TI - [Recent advance in pulmonary function tests--advance in pulmonary function tests for diagnosis and management of asthma]. AB - Development of new machine and small-sized equipment have the advantage on both diagnosis and management of various diseases. In terms of bronchial asthma, the understanding of pathophysiology has been changed from a disease with acute episodes of bronchoconstriction to a disorder with chronic airway inflammation. To verify bronchial responsiveness induced by chronic inflammation, a direct writing respiratory impedance method(Astograph) is more useful compared with a conventional standard method with measuring FEV1.0. Peak expiratory flow(PEF), an index of pulmonary function test with effort, can be measured with peak flow meter which is small and handy. Repeated measurements of PEF have been recommended by the International Consensus Report on diagnosis and treatment of asthma. The PEF monitoring is effective not only on the understanding of individual pathophysiology but also on the long-term management of patients with asthma. It is needed to develop noninvasive simple technique to evaluate airway inflammation although many investigators have examined hypertonic saline-induced sputum or bronchial mucosal biopsy. Repeated measurements of exhaled nitric oxide may become to a safe and valid method to access inflammatory change in the airway. PMID- 11391946 TI - [Recent advances in investigations into brain function and its clinical application]. AB - Recent advances in investigations into brain function and its clinical application are described. The investigations were divided into three method groups consisting of the examinations of; 1) brain electric activity; 2) imaging techniques on activated brain tissue; and 3) collation of the metabolic information on the area of brain focused on. The first group included electroencephalogram(EEG), dipole tracing(DT) and magnetoencephalogram(MEG). The second one, single photon emission computed tomography(SPECT), positron emission tomography(PET) and functional magnetic resonance imaging(fMRI), and the third, magnetic resonance spectrometry(MRS). Here I overview these examinations and report some cases diagnosed with these technologies. PMID- 11391947 TI - [The development of the endoscopic treatment]. AB - With development and improvement of the endoscope equipment, the role of the digestive system endoscope is developing to the application from the pickup of mere lesion to precise diagnosis and endoscopic treatment. The range of the digestive system endoscopic treatment tends to also expand year by year. From the viewpoint of the medicine side, the update of the digestive system endoscope is reported this time. Digestive system endoscopic treatment can be divided roughly into the digestive tract region and the bile pancreas region. There are foreign body extraction, hemostasis methods(peptic ulcer, esophageal varix), excision for the tumor, dilatation methods for the stenosis, etc. as endoscopic treatment in the digestive tract region. In the bile pancreas region, there are endoscopic sphincterotomy, calculus removal of bile duct and ductus pancreaticus, drainage, dilatation methods for the stenosis, etc. The endoscopic treatment develops more and more on both regions with development of together various equipment and improvement in the technique. The invasion to the patient is less than the surgery for the endoscopic treatment, and it is possible to position with the therapy that it retained the function of the organ and was excellent concerning the QOL. It is anticipated with that the effort with the aim of further low invasion will advance in future too. PMID- 11391948 TI - [Virtual bronchoscope system]. AB - In this article, we describe the features of Virtual Bronchoscope System(VBS) and its practical use. VBS is constructed based on 3-D chest CT images. The bronchus region is automatically extracted from 3-D chest CT images by a three-dimensional region growing method. The surface rendering is employed for construction of virtualized tracheo-bronchial tree. It gives us an environment where we can observe inside the bronchi from an arbitrary viewpoint and a view direction. By mouse operation, the user can control the viewpoint and the view direction to fly through inside the airway in real time. VBS is applicable for a variety of purposes such as diagnosis, surgical planning, informed consent, education and training. One of extension of this system is a teaching tool for medical students. In the module for educational use, we have developed four functions for using the system as a teaching tool as follows: (a) automated display of bronchial anatomical names, (b) presenting questions about the currently observed branch in the endoscopic view, (c) display of the path which the user should follow, and (d) display of a question about the location of the artificially created tumor in the bronchus. These functions use the processed results of automated anatomical labeling. The method proposed here combines the knowledge based processing technique 'automated labeling of bronchial branch' and the novel visualization technique 'virtual bronchoscope'. This is one of new teaching tools of medical images. We conclude that this virtual bronchoscope system might have an important role in the medial students' education. PMID- 11391949 TI - [Involvement of interleukin-4, and -13 in the pathogenesis of allergic diseases]. AB - Allergic diseases are immunologic disorders in which various kinds of cells and many mediators are involved. Among them, the importance of interleukin(IL)-4 and IL-13 has recently emerged, based on the analyses of cytokine expression profiles in lesions, model mice, genetic factors, and responses to newly developed reagents. It is known that IL-4 and IL-13 are involved in the pathogenesis of allergic diseases, particularly bronchial asthma, by inducing class switching in B cells and expansion of Th2-type cells. However, IL-4 and IL-13 have a pivotal role in causing bronchial asthma by acting directly on bronchial tissues. Furthermore, it has been demonstrated that a single nucleotide polymorphism in the IL-13 gene is genetically correlated with bronchial asthma. These analyses provide new information for developing better methods of diagnosing and treating bronchial asthma. PMID- 11391950 TI - [Effects of adjuvants on the function of antigen presenting cells in the production of IgE in mice]. AB - In an examination of the effects of adjuvants on the production of IgE, amounts of mRNAs for cytokines in antigen presenting cells(APCs) were assayed by RT-PCR and expressions of surface molecules on the cells were analyzed by flowcytometry when primed with antigen plus adjuvant. When mice were primed with ovalbumin plus alum, the levels of total and specific IgE were higher than those of mice primed with ovalbumin plus CFA. The APCs from mice primed with ovalbumin plus alum expressed higher levels of IL-1 mRNA than those in APCs from mice primed with ovalbumin plus CFA. B7 molecules were more expressed on the surface of the APCs from mice primed with ovalbumin plus CFA. The results suggested that these modulations of the functions of APCs affected the induction of helper T cell subsets in mice primed with different adjuvant. PMID- 11391952 TI - [Positive ratio of allergen specific IgE antibodies in serum, from a large scale study]. AB - Incidences of allergen specific IgE antibodies in a large Japanese population(n = 4,797,158) were studied. Sera from patients were collected in our laboratories during 1994-1998, and assayed with Pharmacias CAP system FEIA. Values greater than 0.70 UA/ml(Score 2) were considered positive. Japanese cedar pollen showed the highest IgE response(positive ratio), followed by house dust, and dust mite. Among food allergens, apple had the highest response, followed by sesame seed, egg white, potato, and tomato. IgE response against Anisakis was significantly higher than that against seafood. Positive ratio varied significantly among regions and among seasons. For example, Hokkaido, compared to other regions, showed lower values for cedar and cypress, but higher for silver birch. Values used in this study were from patients who chose IgE testing, and thus do not represent the general population. However, our findings include positive ratios of rare allergens, and are useful for clinicians who do allergen testing. PMID- 11391951 TI - [Eosinophils and related chemokines]. AB - Chemokines such as RANTES, eotaxin, MIP-1 and MCP-4 are considered to be involved in the pathophysiology of allergic inflammation because of their ability to drive eosinophils through their binding sites, chemokine receptors, expressed on eosinophils. Among those chemokines, RANTES and eotaxin are considered to play important roles in the process of the maturation, migration and activation of eosinophils. An overview of the effect of chemokines on eosinophils throughout their migration from bone marrow to the inflammatory focus is described in this paper. Furthermore, our observations on the effects of chemokines on eosinophils such as adherence through beta-2 integrin, the production of reactive oxygen species, intracellular EG2 content and production of RANTES by eosinophils are reported. PMID- 11391953 TI - [Present situation and problems in allergy testing in vitro]. AB - The present situation and problems related to in vitro assays for measurement of specific immunoglobulin E(IgE) are described. A comparison of four commercially available assay kits(CAP, AlaSTAT, MAST and QAS) revealed that of the allergens studied, common ragweed, mugwort and fungus(aspergillus, candida and alternaria) showed low correlation coefficients. The precise reason for this discordance is not clear, however, we speculated that difference in the origin and extraction method of allergens may affect the correlation results. Random between-the-lot difference was noted: the results for egg white and milk in QAS showed differences of two classes, although the total imprecision expressed in CV(%) was less than 12% when only a single lot was tested. These results suggest that the specific IgE assay may need an improvement in reagents(approval of standard materials and establishment of standard procedures for allergen extraction) and careful daily quality control by the user. PMID- 11391954 TI - [Influences of uncommon isoenzymes on determination of alkaline phosphatase activity by dry-chemistry analyzers]. AB - Dry-chemistry(DC) analysis may be influenced by some matrix effects for measuring uncommon isoenzyme forms. Placental and intestinal alkaline phosphatase(AP) are overestimated by the VITROS DC, compared with results obtained with the wet chemistry(WC) method of Bretaudiere, et al. using 2-amino-2-methyl-1-propanol (AMP) buffer, however, no such discrepancy between AP results in any DC method and that with a routine WC method recommended by Japanese Society of Clinical Chemistry in that 2-ethylaminoethanol(EAE) buffer is used, has been demonstrated. The type of buffer used affects differently the rates of AP isoenzymes activities. We therefore examined whether the presence of uncommon AP isoenzyme forms in serum caused aberrant DC results for AP in comparison with a routine WC method using EAE buffer. Here, serum samples with only liver AP and bone AP(n : 32); high-molecular-mass AP(n : 11); placental AP(n : 12); intestinal AP(n : 13) and immunoglobulin (Ig) bound AP(n : 12) were analyzed for total AP activity on three different DC analyzers: VITROS 700XR, FUJIDRYCHEM 5000, SPOTCHEM 4410 and a WC analyzer: HITACHI 7350. Values obtained in all of the DCs for sera containing only liver/bone AP agreed with those with the WC method. For sera containing placental AP, the VITROS values were higher than those with the WC method, while the FUJIDRYCHEM values and the SPOTCHEM values were lower. The VITROS values and the FUJIDRYCHEM values for sera containing intestinal AP were lower than those with the WC method, while the SPOTCHEM values were higher. All of the DCs did not affect high-molecular-mass AP and Ig bound liver/bone AP types of macro AP, but underestimated Ig bound intestinal type. Ig bound intestinal AP may be sieved by DC multilayer elements. PMID- 11391955 TI - C-reactive protein (CRP) in the cardiovascular system. AB - CRP (C-reactive protein) is an acute-phase reactant, the levels of which increase dramatically in response to severe bacterial infection, physical trauma, and other inflammatory conditions. CRP is found in human atherosclerotic lesions. Atherosclerosis is clearly multifactorial in origin, and chronic inflammation is an important component in its pathogenesis. Focus on inflammation is critical in research on atherosclerosis. Elevated levels of CRP have been associated with increased risk of future coronary artery disease (CAD) events. I have summarized the recent literature on CRP studies in CAD. Both coronary heart disease and dilated cardiomyopathy(DCM) result in congestive heart failure due to myocardial damage. The inflammatory state produced by myocarditis of viral or other origin may induce advanced myocardial damage, resulting in heart failure with a poor prognosis. Routine CRP measurement proved to be valuable for identifying high risk patients with DCM and lymphocytic myocarditis. I suggest that measurement of circulating CRP would be useful for the diagnosis of and for selecting therapeutic strategies for cardiovascular disorders. PMID- 11391956 TI - [Whole blood flow cytometry for detection of activated platelets. I. Platelet identification, blood collection and storage]. AB - In order to assess platelet activation by flow cytometry and activation-dependent monoclonal antibodies, a minute volume(2.5 microliters) of citrated whole blood was mixed for 15 minutes with a cocktail of monoclonal antibodies(MoAb), including PAC1, (a MoAb specific for fibrinogen receptors), and a MoAb against CD62P, (an alpha-granule membrane protein which associates with platelet surface membranes when platelets are activated). After fixation with 1% formaldehyde, the percentage of platelets positive for PAC1 and/or MoAb-CD62P was measured by flow cytometry. Even in resting platelets without stimulation, about 15% of platelets were found to be PAC1-positive and 0.6% of platelets were CD62P-positive. If there was a one-minute delay after needle puncture and before blood collection, the number of PAC1-positive platelets was increased compared to that of blood obtained immediately after puncture. If blood was allowed to stand at room temperature, there was a gradual increase in the number of PAC1-positive platelets, and after 60 minutes, this increase became statistically significant. The addition of Iloprost, (a prostacyclin analogue), immediately after venipuncture did not completely prevent the increase in PAC1-positive platelets after 60 minutes. If the inhibitor was added after first incubating for 60 minutes, however, the percentage of PAC1-positive platelets was reduced to preincubation value. Although flow cytometry is a simple and powerful tool to assess platelet activation, there remain several methodological problems to be resolved before this method may be employed in routine clinical use. PMID- 11391958 TI - [Acute encephalitis and encephalopathy in children--current topics and future strategy]. PMID- 11391960 TI - [Dysplasias of cerebral cortex: basic and clinical aspects]. PMID- 11391959 TI - [Spinal muscular atrophy]. PMID- 11391961 TI - [Frontiers of clinical and basic research of muscular dystrophy: elucidation of responsible genes and their application to molecular therapy]. PMID- 11391962 TI - [The current status and problems of the unauthorized drugs in pediatric neurology]. PMID- 11391963 TI - [Exchange program with Asian and Oceanian researchers]. PMID- 11391964 TI - [Intrauterine and perinatal environment and neurological disorders]. PMID- 11391965 TI - [Bone marrow transplantation of central nervous system diseases]. PMID- 11391966 TI - [Clinico-neurobiological features and therapy in autism]. PMID- 11391967 TI - [Latex allergy in patients with severe motor and intellectual disabilities syndrome]. AB - Our experience of an anaphylactic reaction to latex in a severely disabled patient led us to investigate latex allergy in 58 cases with severe motor and intellectual disabilities syndrome. Latex specific IgE, total serum IgE, and eosinophil counts in peripheral blood were evaluated, as well as past history of treatment with medical latex materials, operation and allergic disorders. Only one case who had been operated three times, had anaphylactic reaction and mild atopic dermatitis. Fifteen cases (25.9%) had latex specific IgE of class 2 or more. In this latex positive group, past history of allergic disorders, such as bronchilal asthma, atopic dermatitis and drug eruptions, were the most prominent factor. Those without such a history were characterized by frequent use of medical latex materials and multiple operations. In conclusion, the danger of an anaphylactic reaction to latex should be recognized, particularly in the medical care for disabled patients. PMID- 11391968 TI - [Assessment of general movements at routine medical examination of one-month-old infants]. AB - On routine medical examination of one-month-old infants, general movements (GMs) were video-recorded for about 5 minutes of 252 infants born at Kyoto City Hospital or other hospitals, including the infants discharged from the NICU. Their ages ranged from 39 postmenstrual to 8 post-term weeks. One of the experimenters assessed 181 infants among 252 available for assessment of their GMs twice at intervals of more than one month without being informed about their clinical histories. The subjects included 19 low-birthweight infants, and their average age was 3.5 post-term weeks. The assessment was carried out in comparison to the "gold standard" GMs depicted in a demonstration video produced by the GMs Trust. The Kappa value for intra-observer concordance was 0.85. Twenty (11.0%) out of the 181 infants were judged abnormal because of their poor repertoire of GMs in both the first and second assessment. Neither cramped synchronized GMs nor chaotic ones were observed. Meanwhile, 155 (85.6%) out of the 181 infants were found to be normal. The incidence of abnormal GMs was significantly higher among infants with history of asphyxia and/or respiratory distress than among low-risk infants. There of the experimenters independently assessed GMs of 50 infants among 181 which had been video-recorded between June and September 1998. The inter-observer concordance was higher among the low-birth weight infants than among full-term ones. In conclusion, qualitative assessment of GMs by an experienced observer at routine medical examination of one-month old infants is a useful method for the follow-up of high-risk infants. PMID- 11391969 TI - [Life-sustaining mechanical ventilation care for children with progressive or degenerative brain disorders]. AB - In the past 10 years, we have treated 25 patients with chronic respiratory insufficiency due to a progressive or degenerative brain disorder. Ten patients died and the other 15 survived. Five of the former and 12 of the latter received life-sustaining mechanical ventilation care. Even in the terminal stage of progressive or degenerative brain disorders, patients can survive for a longer period than previously, if life-sustaining mechanical ventilation care is given. In Japan we do not have a guideline for medical indication or decision-making for children with progressive or degenerative brain disorders. Whenever we see such patients, we have great difficulty in making a decision. It may therefore be necessary to discuss whether we should have such a guideline. PMID- 11391970 TI - [Pathogenesis of acute encephalopathy with shock syndrome]. AB - We presented 3 cases who exhibited important clinical or histopathological findings relevant to the pathogenesis of acute encephalopathy with shock syndrome (AESS). Hyperpermeability of the blood vesseles in the central nervous system was revealed by autopsy in case 1 and by enhancement CT scan in case 2, which implicated that the primary lesion of AESS was vascular damage. Although other cytokines than interleukin (IL)-6 were not evaluated, high value of IL-6 in case 3 might suggest that the some highly activated cytokines caused AESS. PMID- 11391971 TI - [Multiple sclerosis with consciousness disturbance: a case report]. AB - We report here a nine-year-old girl with multiple sclerosis having consciousness disturbance at admission. Neurological examination revealed drowsiness, unstable emotion, decreased visual acuity, disturbance of convergence, and clumsy coordination movements. Her cerebrospinal fluid IgG and myelin basic protein were increased. Electroencephalogram showed intermittent, high voltage slow waves predominant in the frontal lobes. Magnetic resonance imaging (MRI) found multiple demyelinating plaques in the brainstem, thalamus, periventricular white matter. The brainstem reticular formation was involved. Since she had had bilateral acute optic neuritis and papillitis two years before the admission, the diagnosis of multiple sclerosis was made. Methylprednisolone pulse therapy improved her neurological symptoms and MRI findings. Multiple sclerosis in children, unlike that in adults, may present with symptoms mimicking an encephalopathy. Our case suggested that consciousness disturbance in childhood multiple sclerosis results from lesions in the brainstem activating reticular formation including the thalamus. PMID- 11391973 TI - [Mitochondrial DNA T to G mutation 8993 in Leigh encephalopathy and organic aciduria]. AB - We report a 10-month-old female infant with Leigh encephalopathy caused by a T to G mutation at nucleotide 8993 of mitochondrial DNA. Initial manifestations were diarrhea and pyrexia, followed by disturbance of consciousness. Blood chemistry showed lactic acidosis, and cranial T2 weighted magnetic resonance imaging demonstrated symmetric high-intensity areas in the basal ganglia, consistent with Leigh encephalopathy. Analysis of urinary organic acids revealed a increase of alpha-ketoglutamate. Derivatives of branched chain amino acids, which accumulate in maple syrup disease, were also increased. Lipoamide dehydrogenase (E3) deficiency was initially suspected; however, normal activity of pyruvate dehydrogenase complex excluded the diagnosis. The organic aciduria disappeared after two weeks. The CNS lesions in our case were observed more prominently in the floor of the bilateral frontal lobes than in the globus pallidus and putamen. In this case, mitochondrial DNA mutation may have caused organic aciduria and the atypical imaging findings. PMID- 11391972 TI - [Interleukin-6 in the cerebrospinal fluid of two patients with herpes zoster meningitis]. AB - Interleukin-6 (IL-6) levels in the cerebrospinal fluid (CSF) and serum were measured in two immuno-competent children with herpes zoster meningitis, who had vesicles, fever, headache and vomiting before admission. The causative agent was identified as varicella zoster virus (VZV) by detecting an increased antibody index in the serum and specific DNA (by polymerase chain reaction) in the CSF. Both patients fully recovered after treatment with acyclovir. The CSF IL-6 levels were high (260.1 pg/ml, 106.1 pg/ml) at the acute stage and thereafter showed a rapid recovery. The serum IL-6 levels were normal. The increased IL-6 level in the CSF may reflect intrathecal inflammatory response following invasion of VZV into the central nervous system. PMID- 11391974 TI - [The effect of risperidone on the self-mutilation of Lesch-Nyhan syndrome]. PMID- 11391975 TI - [Intranasal midazolam for prevention of status epilepticus]. PMID- 11391976 TI - [Current status and controversies on the management of cerebral main trunk occlusion]. AB - Management of the cerebral main trunk occlusive disease has proceeded during last decade owing to remarkable progress of diagnostic and interventional neuroradiology. Several large clinical trials have established treatment guidelines in most of stroke management. Concerning cerebral main trunk occlusion, management of acute embolic ischemia and surgical indication for chronic carotid artery occlusion has still remained unsettled. In this article, outline of previous discussion and current status and controversies are reviewed. PMID- 11391977 TI - [Escape mechanism of intracellular parasitic bacteria and prospect of new approach to infection control]. AB - Intracellular parasitic bacteria are capable of escaping from the intracellular killing inside macrophages by virtue of highly sophisticated molecular mechanism. Because of the escape from phagocytic defense, infection caused by these bacteria is difficult to be controlled even in the presence of normal host defense system. In order to understand the mechanism of intracellular parasitism of particular types of bacteria, the basic mechanism of intracellular killing inside phagocyte and the strategy of escape by some representative intracellular bacteria are reviewed. Based on the mechanism of T-cell dependent protective immunity, some possible approaches to the infection by intracellular bacteria, especially to tuberculosis, have been discussed. PMID- 11391978 TI - [Congenic rats for hypertension studies]. AB - A congenic rat has a small genomic fragment transferred from a donor strain into a recipient strain. It is therefore possible to make a hypertensive congenic strain with a single unknown hypertensive gene originating from a genetic hypertensive model rat such as SHR. Such a congenic strain is useful in the search for hypertension genes in rats. It can be applied (1) to further dissection of QTL regions, (2) to determination of the boundaries of QTL regions, which is necessary for selection of positional candidate genes, and (3) to physiological and biochemical studies to get intermediate phenotypes. PMID- 11391979 TI - [The Japanese new guideline for the management of hypertension--background of its preparation and characteristics of the new guideline]. AB - The Japanese society of hypertension organized guidelines subcommittee for guidelines for the management of hypertension by request of Japanese physicians and people in the Ministry of Health and Welfare in 1998. Professor Masatoshi Fujishima in the School of Medicine, Kyushu University became a chairman in the committee. He had proceeded the preparation smoothly, and in July, 2000 the new guideline for the management of hypertension was published. The Japanese new guideline for the management of hypertension is slightly different from those in other countries, such as JNC-VI or the WHO/ISH guideline. The Japanese new guideline was prepared under the consideration of characteristics of hypertension in Japan, Japanese life style and recent trends in the management of hypertension in Japan. It is desired that Japanese physicians treat hypertensive patients according to this new guideline, and questionable parts in the guideline should be corrected to make it more useful and valuable. Especially the parts peculiar to the Japanese guideline should be ascertained by further studies in Japan. PMID- 11391980 TI - [Genetic analyses of essential hypertension]. AB - Recent genetic studies using nonparametric linkage analyses have suggested that several chromosomal regions linked with hypertension. On the other hand, statistical estimation implied that linkage analyses did not have enough power to identify hypertension-susceptible genes that seemed to have weaker effects on blood pressure than expected previously. Although association analyses using single-nucleotide polymorphisms have been introduced with enthusiasm, it is necessary to solve several questions before we can apply this strategy to identify hypertension-susceptible genes. Not genotyping but phenotyping will become more and more important in the genetic studies of hypertension. PMID- 11391981 TI - [Genetic polymorphisms of the renin-angiotensin system]. AB - Essential hypertension is a multifactorial disease which is evoked by multiple environmental and genetic factors. Recent progress in molecular biology enables us a comprehensive understanding of the molecular pathogenesis of this disease. The genes of renin-angiotensin system such as renin, angiotensinogen, angiotensin converting enzyme and angiotensin receptors genes are all reported as positive linkage to hypertension. Now ongoing progress of the human genome project will further accelerate the molecular studies on the rennin-angiotensin system and hypertension. PMID- 11391982 TI - [Insulin resistance syndrome]. AB - Insulin resistance and hyperinsulinemia have been observed in essential hypertension. The selective impairment of glucose metabolism in skeletal muscle may accompanied hyperinsulinemia and raise blood pressure through sympathetic nervous system and/or renin-angiotensin system activation, renal sodium retention, proliferation of vascular smooth muscle and leptin. Recently, molecular techniques have applied for investigating the mechanisms of insulin resistance. The mutation of insulin receptor gene, changes of muscle fiber composition and muscle blood flow, abnormalities of insulin signal transduction, and TNF-alpha are considered as involvement of insulin resistance in the skeletal muscle. While further study will be necessary to clarify the mechanisms of insulin resistance and hypertension. PMID- 11391983 TI - [Recent trends in studies of the etiology of hypertension: Central nervous system and autonomic nervous system]. AB - Abnormality of autonomic nervous system is one of the important mechanisms in hypertension. Arterial baroreceptor reflex control of heart rate and sympathetic nerve activity is reset to higher level of blood pressure in hypertension. Abnormality of arterial baroreceptors has been studied as the mechanism of resetting in hypertension. However, recent studies have been focused on the role of central nervous system. Many factors in peripheral system also exist independently in the brain, such as the renin-angiotensin system, nitric oxide, endothelin-1. They contribute importantly to regulation of blood pressure. New approaches to examine the central control of cardiovascular regulation are now developing. Such methods will be useful in the study of the role of specific genes in the particular areas within the brain that regulate blood pressure. PMID- 11391984 TI - [Cardiovascular abnormalities as a cause of hypertension]. AB - This review is an attempt to highlight evidence that may implicate the cardiovascular abnormalities in the pathogenesis of hypertension. Many physiological, pharmacological, and biochemical studies have been conducted in in vitro and in vivo systems. Since blood pressure can rise in response to an increase in cardiac output and/or a rise in peripheral resistance, abnormalities may be present in one or more of the multiple factors that affect these two parameters in hypertension. These multiple factors include various neurohumoral factors. Increased levels of various vasoconstrictor neurohumoral factors have been found in patients with hypertension. Vasoconstrictor neurohumoral factors such as catecholamines, angiotensin II, and endothelin-1 induce vascular smooth muscle cells(VSMCs) proliferation and contraction. On the other hand, vasodilator neurohumoral factors such as natriuretic peptides and adrenomedulin inhibit VSMCs proliferation. Both neurohumoral factors mutually interact and develop hypertension. PMID- 11391985 TI - [Recent trends in studies of the etiology of hypertension: Renal mechanism of blood pressure regulation]. AB - The kidney plays an important role in the control of systemic blood pressure by regulating the composition of body fluid and electrolyte and by producing and releasing various vasoactive substances such as renin-angiotensin and prostaglandins(PGs). The kidney possesses an intrinsic mechanism called 'pressure natriuresis' in which an increase in perfusion pressure promotes natriuresis. According to this mechanism, hypertension can be classified as salt-sensitive or non-salt-sensitive based on the response to dietary sodium loading. Thus, a shift of the curve to the right and a decrease in the slope are the two basic abnormalities seen in the pressure-natriuresis relationship in hypertension, with each representing a different pathophysiology. In non-salt-sensitive essential hypertension, glomerular capillary pressure is within the normal range due to elevated preglomerular resistance, while it is elevated in salt-sensitive hypertension. Such differences may be reflected in the rate of progression of renal dysfunction as well as renal pathology. Finally, the mechanism responsible for the altered renal microcirculation involves dysregulation of intrinsic mechanisms(myogenic responses and tubuloglomerular feedback) as well as various local factors, in particular, angiotensin II, nitric oxide, and oxidative stress. PMID- 11391986 TI - [Recent trends in studies of the etiology of hypertension: Endothelium-derived factors]. AB - Endothelial cells play diverse biological roles, such as maintaining vascular tone and structure. Recent progress in vascular biology has revealed that the endothelium releases a large number of vasoactive substances. These substances are divided into two classes: endothelium-derived relaxing factors(EDRFs), and endothelium-derived contracting factors(EDCFs). It has been shown that EDRFs such as nitric oxide(NO) protects vasculature from atherogenic insults, whereas EDCFs, such as endothelin, have opposite effects and participate in the progression of cardiovascular diseases. The exposure of coronary risk factors decreases the bioactivity of EDRFs and increases the release of EDCFs. Endothelial dysfunction caused by imbalance between EDRFs and EDCFs precedes and promotes atherosclerosis by several mechanisms, such as adhesion of monocytes and platelets, increase in vascular permeability, proliferation and migration of smooth muscle cells. In this article, physiology and pathophysiology of these endothelium-derived substances will be reviewed. PMID- 11391987 TI - [Recent trends in studies of the etiology of hypertension: New endocrine regulators of blood pressure]. AB - The literature review reflects new aspects of humoral regulation in hypertension and target-organ damages with special regard to natriuretic peptide system(NPS) and adrenomedullin(AM). NPS and AM are recently discovered regulators which serve as antihypertensive and target-organ protective factors. These peptides have both diuretic and natriuretic properties and a relaxing effect on the vasculature. Moreover, they antagonize the proliferative and hypertrophic stimuli in the vasculature and heart. Recently, progressive technics of molecular biology clearly revealed crucial roles of these peptides for cardiovascular regulation in both normal and pathological states including hypertension and related organ damages. Natriuretic peptides, potentially AM, are new therapeutic tools for heart failure and main targets for further development of new antihypertensive drugs such as vasopeptidase inhibitor. PMID- 11391988 TI - [Management of hypertension in Japan--current state and clinical issue]. AB - The Japanese Guidelines for the Management of Hypertension (JSH2000) have been published in June, 2000, which basically followed the direction of 1999 WHO/ISH and JNC-VI guidelines. Target blood pressures for young or middle-aged hypertensive patients or hypertensives with diabetes are recommended to maintain less than 130/85 mmHg. In contrast, blood pressure control for hypertension in elderly is set taking the subject's age into consideration with systolic blood pressure lower than 140-160 mmHg and diastolic below 90 mmHg. Among hypertensive cardiovascular diseases, stroke is more common while ischemic heart disease is less common in Japanese than in Caucasians. Frequency of hypertension in Japan, which is estimated to be one-fourth of whole population and two-thirds of persons aged 60 years or over, has been declined in recent years, because of increasing treatment of hypertension, resulting in a decrease in stroke mortality and morbidity. However, the number of persons with hypertension controlled to below 140/90 mmHg seems to be about 20 percent of all hypertensives. Therefore, increases in rates of awareness, treatment and appropriate control of hypertension are the important issue for the management of hypertension in Japanese at present state. PMID- 11391989 TI - [Diagnosis of hypertension and outline of hypertension treatment]. AB - In this review first I described the procedures of diagnosis in patients with essential hypertension. Blood pressure level, risk factor for cardiovascular disease and organ damage due to hypertension are essential in evaluating a patient with hypertension at outpatient clinic. Those criteria are based on Hypertension Treatment Guidelines 2000 published by Japanese Society of Hypertension last year. Next I described the outline of treatment of hypertensive patients according to the above guidelines. The guidelines is our interpretation of the world literature and of current opinions together with our largely documented experience based on the management of many hypertensive patients in Japan. The first-line antihypertensive drugs are long-acting Ca antagonists, ACE inhibitors, angiotensin II receptor antagonists, diuretics, beta-blockers and alpha-blockers. PMID- 11391990 TI - [Measures against environmental and life-style problems in the patients with hypertension]. AB - It is well known that the pathogenesis of essential hypertension is multi factorial, and that the environmental and genetic factors and their interaction play important roles on pathogenesis of hypertension. The environmental factors consist of natural environmental factors, such as weather, seasons, temperature, soil and water, and social environmental factors, such as human relationship, socioeconomic status, residence, education, stress and life-style. Life-style modification including salt restriction, body weight reduction, mineral supplementation, aerobic exercise, alcohol restriction, cessation of smoking and release from psychoemotional stress is the basic strategy for antihypertensive therapy. Not only Joint National Committee (JNC VI) and WHO-International Society of Hypertension (WHO-ISH 1999), but also Japanese Society of Hypertension (JSH 2000) are recommending the life-style modification for the initial treatment of hypertension. PMID- 11391991 TI - [Pharmacological characteristics and choice of antihypertensives]. AB - Many antihypertensive drugs are available to treat patients with essential hypertension. Japanese Society of Hypertension (JSH) recently published 'Guidelines for the management of Hypertension (JSH 2000)'. JSH 2000 chooses six drugs(calcium antagonist, angiotensin converting enzyme inhibitor, angiotensin II receptor antagonist, diuretic, beta-blocker, alpha-blocker) as first lines of antihypertensive treatment. These six drugs have different pharmacological characteristics and there are compelling indications for specific drugs in certain clinical conditions such as diabetes mellitus, organ damages, and elderly hypertensives etc. Usually a low dose of the initial drug choice should be used. The optimal formulation should provide 24-hour efficacy with a once-daily dose, because compliance is better and control of hypertension is persistent and smooth. PMID- 11391992 TI - [Antihypertensive treatment guideline for elderly hypertension]. AB - Recently published Hypertension Treatment Guideline by Japanese Society of Hypertension (JSH2000) contains several unique suggestions for treatment of elderly hypertension in particular. In general, the therapeutic goal of blood pressure in the elderly is set at a higher level than that in the young. For the patients of age 70 and over blood pressure should be controlled less than 150-160 mmHg. The following drugs are recommended as the first line drug in the elderly hypertension: 1. long-acting Ca antagonists, 2. ACE inhibitors (or angiotensin II antagonists) and 3. small dose of diuretics. Combination therapy should be considered when monotherapy of more than 2 to 3 months is not successful to reduce blood pressure below 150/90 mmHg. PMID- 11391993 TI - [Hypertension in children and adolescents]. AB - Normal blood pressure and hypertension were defined according to age and sex based on the data on Japanese children. When high blood pressure is found, both white-coat and secondary hypertension should be excluded. Subsequently lifestyle modifications should be initiated in children and adolescents with essential hypertension. These modifications include: weight reduction, reduction of dietary salt intake, high dietary potassium intake and increased physical activity. When nonpharmacologic treatment is not effective after 3 to 6 months, or when there is an evidence of target organ injury, antihypertensive drugs such as ACE inhibitors and Ca antagonists will be started to control blood pressure. Lifestyle modifications are also important for primary prevention of hypertension in normotensive children. PMID- 11391994 TI - [Hypertension in women]. AB - There are some differences between hypertension in men and women. For example, female sex hormones protect against the development of essential hypertension in premenopausal women. On the other hand, in pregnant women, hypertension must be identified as severe disease in every cases. Essential hypertension in postmenopausal women may be categorized for common disease in many cases. In this article, I show some datas and my comments about female hypertension for mainly daily therapeutic observation. PMID- 11391995 TI - [Secondary hypertension]. AB - In patients with hypertension and chronic renal parenchymal disease, BP should be controlled to 130/85 mmHg or lower (125/75 mmHg) in patients with proteinuria in excess of 1 g/day. Reducing dietary sodium (< 7 g/day) and protein (< 0.6-0.7 g/kg) helps control high BP and renal function in patients with renal insufficiency. As first antihypertensive drug, ACE inhibitors or long-acting Ca antagonists are recommended. In patients with renovascular hypertension, angioplasty is the first choice increasingly to be accompanied by stenting, and surgical revascularization is the next choice. As antihypertensive drugs, beta blockers, ACE inhibitors, and AII-receptor blockers are recommended. Hypertension accompanied by endocrine disease with adenoma or tumor is almost cured or improved by surgical removal. Spironolactone and Ca antagonists are used in patients with idiopathic aldosteronism (bilateral hyperplasia). Alpha and beta blockers are used in patients with pheochromocytoma during preoperative period. PMID- 11391996 TI - [Antihypertensive therapy in patients with stroke]. AB - Most patients with acute ischemic stroke do not need antihypertensive therapy, because the rapid lowering of blood pressure (BP) may reduce cerebral blood flow due to impaired cerebral autoregulation. In patients with severe hypertension, or associated with other complications (hemorrhagic transformation, myocardial infarction, renal failure or dissection of the aorta), antihypertensive therapy should be done cautiously. In chronic phase, the optimal BP level for the prevention of stroke recurrence remains unclear. The presence of the J-curve phenomenon is still controversial. The several large scale trials are now in progress to determine the optimal BP level for the secondary prevention in stroke patients. PMID- 11391997 TI - [Management of hypertension in patients with coronary artery disease]. AB - The management of hypertensive patients with coronary artery disease (CAD) should be started with lifestyle modification for reduction of coronary risk factors. As the principle of drug therapy, rapid decrease in blood pressure below the limits of coronary auto-regulation must be avoided. Clinical trials showed J-curved relationship between diastolic pressure and mortality was not found above 75 mmHg. The current desired goal of blood pressure in patients with CAD might be suggested as 125-140/85-75 mmHg. Pulse pressure should be recognized as a strong predictor for CAD as well as systolic and diastolic blood pressure, especially in elderly patients. The choice of drug therapy have to be based on efficiency in clinical trials, side effects, and also particular condition of each patient. PMID- 11391998 TI - [Therapeutic strategy for progressive renal disease]. AB - Renal disease constitutes an important determinant of cardiovascular disease. Although the mechanisms for the progression of renal impairment remain fully undetermined, available evidence indicate that renal glomerular hypertension is responsible in part for the development of renal injury. In renal disease, afferent arteriolar tone is reported to be reduced, while the augmented intrarenal angiotensin II serves to act as an efferent arteriolar constrictor, both of which result in an increase in glomerular capillary pressure. Angiotensin converting enzyme inhibitors (ACE-I) are established as the agent possessing both antihypertensive and renoprotective actions, which exert vasodilator action on efferent arterioles. Calcium antagonists are also reported to have salutary effect on renal disease, although their beneficial action varies depending on the antagonists used and the underlying disease. The use of calcium antagonists, however, is mandatory particularly under the circumstance where renal failure moderately to severely progresses and the ACE-I cannot be used. PMID- 11391999 TI - [Antihypertensive treatment in diabetics]. AB - Based on many epidemiological studies, hypertensive diabetics have been regarded as high risk for cardiovascular disease. The sixth Joint National Committee guideline (JNC VI) recommended to start hypotensive agents even at high normal blood pressure, i.e. over 130/85 mmHg and lower their blood pressure to less than 130/85 mmHg. If patients are associated with renal dysfunction (proteinuria of more than 1 g/day), the target blood pressure would be less than 125/75 mmHg. These target blood pressures were supported Hypertension Optimal Treatment study and/or UKPDS. ACE inhibitors, Ca-antagonists or alpha-blockers were regarded as one of the first line drugs. Japanese Society of Hypertension has established the guideline for hypertension treatment, in which almost the same recommendation has been made for diabetics except one point, i.e., at high normal blood pressure, lifestyle modification might be the initial treatment followed by hypotensive agents if their blood pressure dose not go down to less than 130/85 mmHg during the next 3-6 months. Based on these guidelines, more vigorous antihypertensive treatment would be recommended in diabetics to reduce micro- and macrovascular disease. PMID- 11392000 TI - [Hyperuricemia in hypertension]. AB - Hyperuricemia is one of the common complications in hypertension. The presence of hyperuricemia is closely correlated with the initiation of hypertension. In addition, the hypertension is known to increase the rate of hyperuricemia. Recently, it has been recognized that the hyperuricemia is one of the risk factors for cardiovascular events in hypertension. Therefore, the control of hyperuricemia might be critical for treatment of hypertension. Although the diuretics are important for the treatment of chronic heart failure and hypertension, it is reported to induce the hyperuricemia. Therefore, the careful and combination therapy for diuretics might be necessary to prevent the treatment induced hyperuricemia. In the treatment of hypertension, especially with hyperuricemia, it might be important to select the drug, which does not influence or reduce the concentration of uric acid. PMID- 11392001 TI - [Treatment of hypertension in the patients with obesity]. AB - Obesity is often accompanied with hypertension and increases cardiovascular events. Japanese new guideline on identification of obesity includes a modified BMI categories and a method of detection of visceral fat obesity in Japanese. Hyper-insulinemia and leptin released from adipose tissue play an important role in the development of hypertension in obese patients. Insulin and leptin increase sympathetic tone which results in sodium retention and hyper-responsiveness of blood vessels. As leptin has also a direct vasodilative and diuretic action, its effect on blood pressure is bidirectional. Life style modification, especially diet and physical exercise are important to obtain the body weight loss and the improvement of insulin resistance. Dynamic exercise at the level of fifty percent of max VO2 for 30 to 60 minutes over three times a week should be recommended for hypertensive patients with obesity. ACE inhibitors improve the hypersympathetic tone and impaired insulin sensitivity in obese patients. Calcium antagonist is also useful for these patients. PMID- 11392002 TI - [Treatment of hypertension with dyslipidemia]. AB - Lifestyle modifications are the first approach to the treatment of dyslipidemia and hypertension, that is, control of overweight; reduced intake of saturated fat, cholesterol, sodium chloride, and alcohol; and increased physical activity. In high doses, thiazide diuretics and loop diuretics can induce at least short term increases in levels of total plasma cholesterol, triglycerides, and low density lipoprotein cholesterol. Low-dose thiazide diuretics do not produce these effects. beta-blockers may increase levels of plasma triglycerides transiently and reduce levels of high-density lipoprotein cholesterol. alpha-blockers may decrease serum cholesterol concentration to a modest degree and increase high density lipoprotein cholesterol. Angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers, calcium antagonists, and central adrenergic agonists have clinically neutral effects on levels of serum lipids and lipoproteins. PMID- 11392003 TI - [Nasal CPAP treatment and hypertension and altered cardiovascular variability associated with obstructive sleep apnea (OSA)]. AB - In the past 5 years several epidemiologic studies have demonstrated that sleep related breathing disorders are an independent risk factor for hypertension, probably resulting from a combination of repetitive episodes of hypoxia, hypercapnea, arousals, and a striking surge in sympathetic excitation, and altered baroreflex control during sleep. Obstructive sleep apnea (OSA) may lead to the cardiac arrhythmias and myocardial ischemia and it is a possible risk factor for stroke. We confirmed that nasal CPAP has been shown to lower blood pressure in some hypertensive OSA patients. Early recognition and treatment of sleep-apnea may improve cardiovascular function. PMID- 11392004 TI - [Antihypertensive drug therapy: adverse effects and drug interactions]. AB - The number of classes of antihypertensive drugs has been growing. The options of choice of the drugs now are so great that it seems not to be difficult to reduce blood pressure effectively. Newly developed classes have been shown to provide beneficial effects in variety of hypertension-related processes. However, each drug shows significant differences in adverse effects, incidental properties and interactions with non-antihypertensive drugs. To improve quality of life, compliance with medication, and beneficial effects on pathophysiological and cardiovascular risk conditions of patients, we should be well acquainted with adverse effects and possible disadvantages produced by antihypertensive drugs in particular situations. PMID- 11392005 TI - [Comparison between AngII receptor antagonist and ACE inhibitor]. AB - Angiotensin II(AII) accelerates the progress of cardiovascular diseases. This was proved by the fact that the blockade of renin-angiotensin system provided clinical benefits for patients with cardiovascular diseases. This review focuses on the differences between AT1-receptor antagonist and ACE inhibitor in basic and clinical aspects. Beside decreased AII concentration, increased tissue bradykinin concentration may contribute to the beneficial effect of ACE inhibitor, on the other hand, this increases the rate of cough to decrease the compliance. Increased AII concentration by AII receptor antagonist may antagonize the binding of the drug as well as stimulate AT2 receptor subtype. ACE inhibitor can not block the effect of non-ACE AII formation, but AII receptor does. These differences should be considered for their clinical use. PMID- 11392006 TI - Long descending lymphatic pathway from the pancreaticoduodenal region to the para aortic nodes: its laterality and topographical relationship with the celiac plexus. AB - In 6 of 15 postmortem-treated cadaveric specimens, we found macroscopically thick lymphatic collecting vessels that originated from not only the nodes along the common hepatic artery (No. 8 nodes) but also from the pancreaticoduodenal region, and which drained directly into the para-aortic nodes immediately below the left renal vein (No. 16b1-inter or -latero nodes). The collecting vessels, if they originated from the ventral (dorsal) visceral side, passed to the left (right) of the superior mesenteric and celiac arteries. Moreover, the right-side vessels (5 specimens) were classified into superficial and deep courses to the celiac plexus, whereas they were superficial in the left side (2 specimens). One of the deep (right) courses continued to the thoracic duct without any intercalated nodes. In addition, another deep route drained into the para-aortic node immediately above the left renal vein (No. 16a2-inter node). We consider that these collecting vessels form "direct descending pathways" from the relatively peripheral lymphatics in the upper abdomen toward the thoracic duct origin. The pathway seems to be a collateral, or even major drainage route, and it appears responsible for skipped metastasis of primary cancer. Since the classical, limited entity of the intestinal lymph trunk does not coincide with our pathway, it should be reconsidered. The proposed entity of the direct, long descending pathway will influence the selection and modification of lymphadenectomy methods in cancer surgery in the pancreaticoduodenal region. PMID- 11392007 TI - Potential fascial dome made by the upper leaf of the phreno-esophageal membrane. AB - We describe the configuration and size of the artificial fascial dome created in 57 cadavers. This dome protrudes into the thoracic cavity from the esophageal hiatus. This dome was a potential space realized by finger dissection (i.e., a specific but common surgical procedure during surgery of the upper part of the stomach). The vagus nerves penetrated the top of the dome and ran down along the esophagus. The height of the ventral wall of the dome ranged from 10-60 mm, while the dorsal wall was 10-40 mm longer than the ventral one since the dorsal wall attached to the lower, dorsal limb of the esophageal hiatus. Accordingly, the dorsal wall separated the "thoracic" aorta from the "abdominal" esophagus. We considered that the upper leaf of the phreno-esophageal membrane forms the fascial dome, although the lower leaf of the membrane was not identified in this study. According to the results, we proposed a schematic representation of the phreno-esophageal membrane. PMID- 11392008 TI - Morphological differentiation of nerve fibers: central, peripheral, myelinated and unmyelinated. AB - We have developed a new technique for the morphological differentiation of various nerve fibers which is especially suitable for the morphometric study of nerve fibers of the human nervous system with the help of an image-analyzer. The knowledge from findings by this technique, which is based on several study methods, may be of importance in promoting further neuromorphologic studies and in properly understanding various aspects of neurological symptomatology and the aging process of the nervous system including nerve fibers. PMID- 11392009 TI - Unmyelinated nerve fibers of the human mandibular nerve. AB - The aim of this research is to find and to evaluate morphometorically the unmyelinated nerve fibers in the human mandibular nerve using a light microscope. Our report demonstrates for the first time the presence of the unmyelinated nerve fibers of the human mandibular nerve stained by a special method. Our results also indicate that there is a morphometric change with aging in the unmyelinated axons of the nerve. PMID- 11392010 TI - Are there one million nerve fibres in the human medullary pyramid? AB - It has been the accepted opinion that there are one million nerve fibres in the human medullary pyramid. This seemed to be confirmed in several old reports. But we cannot agree with this opinion. We made nitrocellulose-embedded sections from three normal male brains, and stained them by our modification of Masson-Goldner method. With this method, myelinated axons appeared in blue, whereas the glial processes were coloured in red, which allowed easy discrimination between the two. After morphometric evaluation of the pyramidal axons under the microscope, it appeared without the slightest doubt, that the number of axons does not exceed one-tenth of one million. PMID- 11392011 TI - The anatomy of the lacrimal portion of the orbicularis oculi muscle (tensor tarsi or Horner's muscle). AB - Horner's muscle (the palpebral part of the orbicularis oculi muscle) has a fan shaped origin in the lacrimal bone. Its muscle fibers are oriented from 160 to 210 degrees relative to the ear-eye plane and converge towards the medial palpebral commissure. Then the muscle divides into superior and inferior bundles of fibers. Some of the lower fibers participate in the formation of the superior bundle and some of the higher fibers participate in the formation of the inferior bundle and, thus, some of Horner's muscle is twisted. Each bundle courses laterally to the lateral palpebral commissure and has three insertions. The first insertion is located at the medial margin of the tarsi. The second insertion is into the subcutaneous tissue along the palpebral margins. Minute fascicles of Horner's muscle are fastened to the palpebral margins. The third insertions are into the lateral palpebral ligament and subcutaneous connective tissue of the lateral commissure. Serial histological sections of a fetus at 14 to 16 weeks gestation revealed that the extent of the envelope formed by Horner's muscle around the lacrimal canaliculus decreases gradually from the lacrimal papilla to the lacrimal sac. The various observations suggest the following roles for Horner's muscle: (1) it closes the medial canthus of the eye and closes the lacrimal punctum; (2) it pulls the tarsus medially; (3) it tautens the palpebral margins and presses against the eyeball; and (4) it squeezes the lacrimal canaliculus with a decreasing gradient of pressure from the lacrimal papilla to the lacrimal sac. These actions are likely to be important for the flow of lacrimal fluid in the lateral to medial direction on the eyeball, for maintenance of the thickness of tear film over the cornea, for opening and closing of the lacrimal punctum, and for passage of the lacrimal fluid from the canaliculus to the sac. Horner's muscle appears, thus, to be a muscle of prime importance in all phases of the flow of lacrimal fluid. PMID- 11392012 TI - The laminar structure of the common opossum masseter (Didelphis marsupialis). AB - Using three heads of the common opossum (Didelphis marsupialis), which may be considered to have a primitive mammalian form and therefore be appropriate for this study, the laminar structure of the masseter was investigated. We also attempted a comparative anatomical study of the relationships of food habits to the laminar structures of the masseter, zygomatic arch and mandibular ramus. In the common opossum masseter, a total of six layers, the primary and secondary sublayers of the superficial layer, the intermediate layer, and the primary, secondary and third sublayers of the deep layer as a proper masseter, were observed. These layers showed a typical reverse laminar structure, with the layers of tendons and muscles alternating. The maxillomandibularis and zygomaticomandibularis muscles were observed in one layer each, as an improper masseter. The laminar structure of the common opossum masseter was shown to be more similar to that of carnivorous placental animals than that of the herbivorous red kangaroo, a similar marsupial. In regard to the number of layers in the laminar structure of the masseter, the results of both this study and those of our predecessors' showed that differences in food habits affect the deep layer in the proper masseter of marsupials and placental mammals, and that of the maxillomandibularis muscle of placental mammals in the improper masseter. PMID- 11392013 TI - IH at the crossroads, Part II. PMID- 11392014 TI - The new AMA ratings. PMID- 11392015 TI - Providing proper equipment. PMID- 11392016 TI - Workplace implementation of AEDs. PMID- 11392017 TI - Tracking IAQ problems to their source. PMID- 11392018 TI - IAQ priorities for mold, yeast, bacteria & spores. PMID- 11392019 TI - Becoming world class in HSE management. PMID- 11392020 TI - Involving the total organization. PMID- 11392021 TI - Putting the pieces together. PMID- 11392022 TI - Update on progestogen therapy. PMID- 11392023 TI - Pharmacokinetics of progesterone administered by the oral and parenteral routes. AB - Progestogens, which include the natural hormone, progesterone, and synthetic steroid derivatives, are used to treat a variety of reproductive disorders. Since synthetic progestogens are often associated with undesirable side effects, improved formulations and modes of administration of progesterone have been sought. In order to find an optimal therapeutic agent, different progesterone formulations and routes of administration have been studied. This paper explains several methods of measuring progesterone levels and summarizes clinical data on the pharmacokinetic profiles of natural progesterone administered by the oral, intramuscular, vaginal, intranasal, percutaneous, sublingual and rectal routes. PMID- 11392024 TI - Progestogen metabolism. AB - There are multiple metabolic pathways for progesterone. Progestogens are metabolized differently as compared with the fate of endogenous progesterone, and the metabolites are significantly dependent upon the route of administration and the physiologic and pathologic state of the patient. Marked variations between patients have also been observed. Many organs metabolize progestogens, each in its own way. In turn, the metabolites each have different biologic potencies, which include major effects on the central nervous system. Compared with the knowledge base about progesterone metabolism, much less is known about medroxyprogesterone acetate and other C-6 and 19-NOR testosterone-derived progestogens. PMID- 11392025 TI - Progesterone and the progesterone receptor. AB - During the 1990s, extensive research has effectively mapped the progesterone receptor-mediated actions of progesterone and has more recently uncovered nonreceptor-mediated effects--the effect of progesterone on uterine sensitivity to oxytocin, for example, involves direct, nongenomic progesterone action on the uterine oxytocin receptor. However, the majority of progesterone effects occur as a result of progesterone-receptor-mediated action, where progesterone behaves as a hormone-dependent transcription factor, probably because these receptors are widely distributed in the body. A distinguishing characteristic of progesterone receptors is the existence of two isoforms, A-form and B-form. In most tissues coexpressing progesterone receptors, estrogen controls the regulation of progesterone receptors, thereby also controlling sensitivity to progestins. Thus, progesterone receptor expression is upregulated by estrogen and downregulated by progesterone in most target tissues. PMID- 11392026 TI - Regulating menstrual bleeding. A prime function of progesterone. AB - Progesterone plays a crucial role in the coordination of endometrial breakdown as well as in the determination and regulation of menstrual blood loss and endometrial transudation of fluid. Furthermore, long-term exposure of the endometrium to progesterone can result in abnormal angiogenesis. Although additional investigations of the estrogen-progesterone relationship are needed, the relative concentrations of these two hormones appear to have important effects on bleeding patterns. Current research is aimed at greater understanding of the ways in which these two hormones interact with several complex cellular and metabolic mechanisms to regulate endometrial bleeding. PMID- 11392027 TI - Genes associated with embryonic attachment and implantation and the role of progesterone. AB - Implantation in humans is a complex process that involves embryo apposition and attachment to the maternal endometrial epithelium, traversing adjacent cells of the epithelial lining and invasion into the endometrial stroma. These processes involve a variety of molecules that are not unique in themselves but play unique roles in the process of implantation. Genes important to embryonic attachment include the epidermal growth factor (EGF) family (EGF, heparin-binding EGF-like growth factor and amphiregulin) and the cytokines (colony-stimulating factor, leukemia inhibitory factor and interleukin-1), as well as a variety of cell adhesion molecules and other glycoproteins. Epithelial factors important in attachment may be regulated by paracrine interactions via the endometrial epithelium and the endometrial stroma, which is a progesterone-responsive tissue. Investigations into genetic knockout animal models and natural mutations in the mouse have demonstrated that genes important to the implantation process affect both embryo attachment and decidualization and include cyclooxygenase-2 and the homeobox gene HOXA-10. Calcitonin is believed to play a role in preparing the apical cell pole for contact with the trophoblast. A number of factors contribute to endometrial regulation by progesterone; some are important in embryo attachment as well as in the invasive phase of implantation. Four specific factors regulated in the endometrial stroma by progesterone are transforming growth factor-beta, interleukin-1 and insulin-like growth factor binding protein 1, tissue inhibitors of metalloproteinases (TIMPs) (especially TIMP-3) and fibronectin, all of which have been demonstrated to inhibit trophoblast invasiveness. Current research should provide answers regarding the effects of various levels of progesterone on the implantation process. PMID- 11392028 TI - Role of progesterone in normal breast physiology. AB - The extant literature indicates that progesterone significantly influences normal mammary growth and differentiation. However, most breast tissue research is conducted using malignant cells, in which the progesterone activity differs greatly from that in normal cells. Although progesterone has been demonstrated to support cyclic proliferation in the breast during the menstrual cycle and pregnancy, in vitro studies have been inconsistent in their assessments of progesterone's role in proliferation. Similarly, mitotic activity in the breast reaches its peak during the progesterone-dominant luteal phase of the menstrual cycle, and therefore some researchers claim that progesterone plays a major role in breast cancer; however, several clinical observations have found evidence dismissing progesterone as a key factor. Thus, researchers seek to expand our current understanding of the role of progesterone in breast physiology. PMID- 11392029 TI - Progestogens and cardiovascular disease. A critical review. AB - Although progestational hormones are clearly beneficial in preventing estrogen induced endometrial hyperplasia and cancer, their effect on other areas is far less clear. Of particular interest is the attenuating effect medroxyprogesterone acetate (MPA) has on the cardiovascular benefits of postmenopausal estrogen treatment. MPA reduces the dilatory effect of estrogens on coronary arteries, increases the progression of coronary artery atherosclerosis, accelerates low density lipoprotein uptake in plaque, increases the thrombogenic potential of atherosclerotic plaques and promotes insulin resistance and its consequent hyperglycemia. These effects may be largely limited to MPA and not shared with other progestogens. PMID- 11392030 TI - Progestogens in the treatment of secondary amenorrhea. AB - Secondary amenorrhea is usually the primary purpose for which manufacturers of new progestogens seek FDA approval. Standards for safe and efficacious treatment of this condition have evolved throughout the past two decades. Studies have evaluated a variety of patient populations, corresponding with the several definitions of secondary amenorrhea that may be found in the literature. Ongoing research seeks to expand the indications for progestogen therapy to include hormone replacement in perimenopausal and postmenopausal women. PMID- 11392031 TI - Endometrial hyperplasia. Risks, recognition and the search for a safe hormone replacement regimen. AB - Endometrial hyperplasia is an acknowledged risk of unopposed estrogen replacement therapy (ERT). Several categories of hyperplasia are now recognized, among which atypical complex hyperplasia is the most likely to progress to carcinoma. Adding a synthetic progestin or natural progesterone to estrogen therapy has been shown to decrease or eliminate the endometrial risk associated with ERT. However, the addition of synthetic progestins has been associated with uncomfortable side effects, reversal of some of the cardiovascular and metabolic benefits of estrogen, and unwanted bleeding. The use of natural micronized progesterone in lieu of synthetic progestins alleviates the former two drawbacks, while careful scheduling of estrogen and progesterone dosing can eliminate the latter. In Europe, where natural progesterone has been in use for some time, a cyclic combined regimen comprising estrogen and 100 mg micronized progesterone administered on days 1-25 has been shown to provide endometrial safety, absence of bleeding and a high rate of tolerability. PMID- 11392032 TI - Use of progesterone in assisted reproduction. AB - Progesterone, supplied in a variety of formulations, is commonly used in assisted reproduction. However, even the commonly accepted uses of progesterone in fertility care are believed to be largely empirical, as are the current available treatment options. While a critical balance of estrogen and progesterone is necessary for successful embryo implantation, achieving that balance is subject to differences in prescribing practice. In evaluating recommendations for the use of progesterone, discussion of its role in assisted-reproduction procedures must distinguish between known therapeutic benefits and empirical benefits. PMID- 11392033 TI - Protecting the endometrium. Opposing the hyperplasia/malignancy potential of ERT. AB - Many trials have examined the clinical and histologic effects of various hormone replacement therapy combinations with the objective of minimizing the incidence of hyperplasia and the potential for subsequent development of adenocarcinoma. Reviewing the results of these trials, it appears that high-dose, long-term progestogen therapy is effective in protecting the endometrium, with duration having a greater impact than dose. Among women given 0.625 mg conjugated equine estrogen (CEE), sequential regimens should include 5 or 10 mg medroxyprogesterone acetate (MPA) or 200 mg micronized progesterone for 12 days or more. Continuous combined regimens require 2.5-5 mg MPA. With women who are taking 1.25 mg CEE the data are less clear, but recommendations include administration with 10 mg MPA for 12-14 days or 5 mg MPA continuous combined therapy. PMID- 11392034 TI - Vaginal bleeding patterns in women receiving hormone replacement therapy. Impact of various progestogen regimens. AB - Postmenopausal bleeding is more prevalent than was previously thought, occurring in women regardless of whether or not they are on hormone replacement therapy. While estrogen-only regimens have been used for several beneficial effects, bleeding patterns associated with these regimens can be irregular and unpredictable, causing discomfort to the patient as well as increasing the risk of both endometrial hyperplasia and carcinoma. Studies in recent years have examined the effects of estrogen-only regimens as compared to different estrogen plus-progestogen combination therapies to help regulate and minimize postmenopausal bleeding while providing endometrial protection. PMID- 11392035 TI - The lack of effect of progestogen on bone. AB - Several critical issues surround the possible effects of progestogens in postmenopausal women. No comparative studies of the effects of estrogen and progestogens on bone loss have been conducted to date, but a review of the available evidence suggests that endogenous progesterone has little effect on endometrial secretory homeostasis. The evidence also suggests that when used alone at normally prescribed dosages, progestogens have neither a direct effect on osteoporosis nor an additive effect when used as a component of hormone replacement therapy (HRT). The positive effects of HRT on bone seen in postmenopausal women are more likely a direct result of the estrogen component. PMID- 11392036 TI - Estrogen, progestogens and cardiovascular risk. AB - This paper provides an overview of hormone replacement therapy (HRT) and its relationship to cardiovascular risk factors, based on several recent studies that evaluated risk using lipid and nonlipid parameters. In the Postmenopausal Estrogen/Progestin Interventions (PEPI) trial, which included a placebo group, an unopposed-estrogen group and three different estrogen-progestogen combination groups, high-density lipoprotein (HDL) cholesterol increased in all treatment groups. Women assigned to conjugated equine estrogen (CEE) alone or CEE plus micronized progesterone (MP) had significantly greater HDL increases than did other treatment groups. Also in the PEPI trial, low-density lipoprotein cholesterol decreased 10-15% in all active treatment groups, regardless of the progestogen evaluated. In the nonlipid measurements of the PEPI trial, fibrinogen was reduced in the nonplacebo groups. Carbohydrate metabolism, measured by two hour postchallenge insulin level, decreased in all groups, including placebo; however, two-hour glucose increased in the CEE plus medroxyprogesterone acetate (cyclic and continuous) groups vs. placebo but was unchanged in CEE alone and CEE plus MP groups. Neither systolic blood pressure nor diastolic blood pressure differed between PEPI groups. Estrogen, with or without progestogen, was associated with a lower weight gain as compared to placebo over the course of the PEPI study. Also discussed in this review are data on oral contraceptive-related cardiovascular risk. Current generations of oral contraceptives have been found to have some deleterious effects on lipids but fewer than those seen with earlier preparations. PMID- 11392037 TI - Progestogens used in menopause. Side effects, mood and quality of life. AB - Progesterone receptors are found in many of the same brain areas as estrogen receptors, including the hypothalamus and limbic system. The limbic system, particularly the amygdala, plays a prominent role in regulating emotion and mood. Progestogens decrease brain excitability, whereas estrogens increase it. This explains, in part, why women with epilepsy have a higher frequency of seizures during the late follicular and ovulatory phases of the menstrual cycle than during the luteal phase. In addition, progesterone has been shown to have profound anesthetic properties and to increase the concentration of monoamine oxidase (MAO), the enzyme that catabolizes serotonin in the brain), whereas estrogen decreases MAO, thereby increasing the concentration of serotonin. The purpose of this paper is to review the extant research regarding these biologic effects of progestogens on brain function. PMID- 11392038 TI - [Treatment of diabetes with the insulin pump]. AB - Diabetes control by subcutaneous insulin infusion (CSII) by an insulin pump becomes the therapeutic option, which most closely mimics the physiologic insulin secretion pattern. It may bring glycemia close to normal levels even in cases where the standard injection insulin therapy fails. After CSII installation the mean daily dose of insulin can be reduced and the compensation of diabetes improves (decreases). CSII is usually installed in diabetic patients where the disease cannot be sufficiently controlled, namely in patients with "dawn phenomenon", severe hypoglycemia, and progression of diabetic complications. Results of long-term follow-up study of CSII-treated diabetic patients at our department (IKEM) confirmed the high efficacy of this method and the minimal incidence of therapy-related complications (severe hypoglycemia, ketoacidosis, body weight increase, technical complications). PMID- 11392039 TI - [Care of patients with the diabetic foot syndrome based on an international consensus]. AB - In the last year an International Consensus on the Diabetic Foot has been developed by a group of independent experts from all over the world. The definition of the diabetic foot is based on WHO criteria as infection, ulceration and/or destruction of deep tissues associated with neurological abnormalities and various degrees of peripheral vascular disease in the lower limb. It is one of the most serious complications of diabetes: approximately 50% of all non traumatic amputations are performed on patients with diabetes, it is a frequent cause of hospitalisation and disability of diabetic patients. Comprehensive prevention and therapy of diabetic foot provided by a podiatric team may reduce the number of amputations by 50% and it may decrease substantially the cost of long-lasting therapy. In all countries foot-care management should be organised at the general practitioner and podiatric nurse level and specialised foot centres with a diabetologist, a podiatric nurse, a radiologist, general, orthopaedic and vascular surgeons, an orthopaedic technician and a physiotherapist should be formed. The podiatric team has the following responsibilities--to identify patients at high risk and monitor them and to treat patients with ulcers. Treatment of diabetic foot must include non-weight bearing, long-lasting antibiotic therapy, improving circulation and topical treatment debridement. PMID- 11392041 TI - [Paired filtration dialysis and free radicals]. AB - BACKGROUND: Extracorporeal dialysis compensating kidney function represents a risk of elevated production of free radicals (FR). Paired filtration dialysis (PFD) is a hemodialysing method used to compensate kidney function. The aim of our work was to study effects of PFD with two types of hemodiafilters on the activity of free radicals. METHOD AND RESULTS: Group of nine regularly dialysed patients was treated with PFD 1) with demodiafilter SG3, composed of polysulphone high-flux hemodiafilter and hemophane low-flux dialysator, 2) with hemodiafilter SG30, composed of identical hemodiafilter as the above on and of a polysulphone low-flux dialysator. Parameters related to FR were examined before, at 30th minute and at the end of procedure. Plasma concentration of substances reacting with thiobarbiturate acid (TBARS) increased at 30th minute when PFD with SG3 was used (3.24 +/- 0.36 versus 3.48 +/- 0.31, p < 0.01) and at the end of the procedure (3.24 +/- 0.36 versus 3.58 +/- 0.48, p < 0.05). Glutathione values in erythrocytes (GSH) decreased at 30th minute of PFD with SG3 (1.85 +/- 0.27 versus 1.68 +/- 0.20, p < 0.05). Plasma antioxidative capacity decreased at 30th minute and at the end of PFD with equal significance when either type of treatment was used (p < 0.001). Glutathione peroxidase (GSHPx) decreased at 30th minute of PFD with SG30 (35.6 +/- 3.8 versus 32.2 +/- 3.1, p < 0.05). Selenium (Se) in blood decreased at 30th minute (44.9 +/- 5.4 versus 40.4 +/- 5.9, p < 0.05) and at the end of PFD with the same membrane (44.9 +/- 5.4 versus 39.4 +/- 5.2, p < 0.05). CONCLUSIONS: Elevation of TBARS and decrease of GSH show the presence of oxidative stress during the PFD treatment with hemodiafilter SG3. Changes probably result from the contact of blood with the hemophane membrane. The decrease of GSHPx during PFD with hemodiafilter SG30 is probably caused by the loss of Se. From the point of FR production, hemodiafilter SG30 can be considered as more advantageous, from the point of Se loss and decreased activity of GSHPx, hemodiafilter SG3 has better effects. PMID- 11392040 TI - [IGFBP-1 and its protective role in the pathogenesis of microangiopathy in type 1 diabetes mellitus]. AB - BACKGROUND: The role of IGF-I/IGFBP's system in the pathogenesis of diabetic vascular complications is widely discussed in the literature. We studied the influence of this system on microvasculature in patients with type 1 diabetes with respect to the effect of IGFBP-1. METHODS AND RESULTS: 17 patients with type 1 diabetes were included in the study. We examined IGF-I, IGFBP-1 and IGFBP-3 serum levels, parameters of compensation of diabetes and basic values of lipid metabolism. The function of microvasculature was examined using the laser-Doppler system Periflux. We didn't found any relation between the total IGF-I serum levels, parameters of lipid metabolism or level of diabetes compensation and the degree of impairment of the function of microcirculation. IGFBP-1 serum levels positively correlated with the peak perfusion in thermal hyperaemia (r = 0.39; p < 0.03). IGFBP-3 serum levels did not affect the function of microcirculation. CONCLUSION: We can conclude that the activity of IGF-I/IGFBP's system belongs to factors contributing to the development of diabetic microangiopathy in type 1 diabetes. IGFBP-1 as the modulator of IGF-I activity plays probably protective role against the progression of microangiopathy. These results correspond with observations of some other authors. PMID- 11392042 TI - [Fire-eater's pneumonia--a difficult differential diagnosis]. AB - Acute aspiration of petroleum or other hydrocarbon products can cause a distinct type of chemical pneumonitis known as fire-eater's pneumonia. Initial findings on chest X-rays are ambiguous and not uniform in nature. Among the most common belong mottled or patchy, local or diffuse perihilar and basal uni- or bilateral infiltrates, areas of atelectasis, tumor-like lesions and rarely pleural effusions. CT finding often shows formation of smaller or larger pneumatoceles. Those cavities tend to regress spontaneously over weeks or months. Clinical diagnosis is sometimes very difficult, especially in cases, when past history of "fire-eating" is not known to the physician. The authors present clinical manifestation, sequential X-ray and CT findings in one patient with fire-eater's pneumonia. To diagnose the disease, not only paraclinical exploration is needed, but also the patient's past history plays an important role. Without it this puzzling clinical unit can remain undiagnosed. PMID- 11392043 TI - [Review of revascularization and arterial reconstructive procedures in arteriosclerosis performed at the Hospital in Lucenec]. AB - The authors present a review of revascularisation and reconstruction operations for arteriosclerosis in a group of 2341 patients (1754 (75%) operations). Direct reconstruction operations were made 825 times (47%), indirect operations- sympathectomy 598 times (34.1%) and endovascular intervention 331 times (18.9%). Intervention vascular radiology significantly changes the indication, the strategy and the tactics of surgical treatment of arteriosclerosis and its complications. PMID- 11392044 TI - [History and perspectives of surgical and endovascular therapy of aneurysms of the abdominal aorta]. AB - The author presents a historical review of the development of surgical and endovascular techniques in the treatment of aneurysms of the abdominal aorta. He describes the decisive cornerstones of treatment of the disease such as the first resection of an aneurysm of the abdominal aorta implemented by Dubost in Paris in 1951, the development of artificial vascular prostheses (vinyon D, dacron etc) and the detection of endovascular techniques by Volodos in Kiev (1986) and Parodi in Buenos Aires in 1990. The author discusses also contemporary pitfalls of endovascular treatment and reflects on indication criteria of the "open" and endovascular technique. PMID- 11392045 TI - [Sudden acute vascular diseases requiring surgery--still a reality]. AB - In the introduction the authors emphasize that despite the fact that acute angiosurgical attacks have a fairly typical symptomatology with a characteristic clinical finding, physicians who participate in their diagnosis and treatment are not aware of it. In the clinical explanation and the material the authors mention the causes of acute angiosurgical attacks and the fact that after acute ischaemia reperfusion occurs. In a summary table they demonstrate the dynamics of the development and increase of acute angiosurgical attacks in 1985 and 1995 in the Slovak Republic, based on statistical data of the Institute of Health Information and Statistics. In the discussion it is emphasized that in the therapeutic results of acute angiosurgical attacks an important part is played by early restoration of the circulation, at least within 6 hours, but preferably within 3 hours after the onset of clinical symptoms. In conjunction with ischaemia and reperfusion attention is drawn to the fact that during early reperfusion in the tissue or organ oxygen radicals are formed which participate in a significant way in reperfusion damage. On account of the complexity of ischaemia and reperfusion in acute angiosurgical attacks it is useful to use the clinical term "acute ischaemic reperfusion syndrome". The authors mention possibilities how this syndrome can be favourably influenced by suitable angiosurgical procedures and pharmacological preparations. In the conclusion the authors emphasize the rising trend of acute angiosurgic attacks, whereby the therapeutic results are not always satisfactory. PMID- 11392046 TI - [Diastematomyelia and the tethered spinal cord syndrome. Case report]. AB - The authors present a patient with diastematomyelia and associated congenital malformations of the spinal cord. They caused tethering of the spinal cord. Patient had cutaneous marks of malformation from birth. The congenital malformation was operated at the age 5 years. MRI and surgical findings confirm the existence of diastematomyelia together with thick tethered filum terminale and lipoma of filum terminale. On surgery the bony septum was removed, the filum terminale was resected and the lipoma was excised. Each of the anomalies caused the tethering of the spinal cord and restricted its physiological ascent. Only careful surgical revision with subsequent removal of all causes of the tethering of the spinal cord produced good result. Diastematomyelia is a congenital malformation with some morphological variations and coexistence of other malformations. The presented case is a typical example of malformations associated with diastematomyelia. PMID- 11392047 TI - [Effect of portosystemic anastomosis on the degree of splenic inhibition]. AB - Secondary hypersplenism is a well-known complication of portal hypertension. Reduced platelet count is a more alarming sign for the physician than risk for the patient. Improvement of thrombocytopenia is urgent, when portal hypertension with splenomegaly and thrombocytopenia presents with life-threatening haemorrhage from gastroesophageal varices. In this case, treatment aimed at stopping the bleeding may be more beneficial than any intervention on the spleen. In this study, we evaluated long-term effects of an elective distal splenorenal shunt or small diameter H-shunt on splenomegaly and thrombocytopenia in 26 patients with portal hypertension operated for repeated bleeding from oesophageal varices. 25 patients had splenomegaly and 16 patients had thrombocytopenia before shunting. Surgery corrected splenomegaly in 16 patients (64%), platelet counts increased in 13 of 16 patients with thrombocytopenia (81.2%). CONCLUSION: Selective or partial portal decompression is sufficient to alleviate thrombocytopenia and splenomegaly associated with portal hypertension. PMID- 11392048 TI - [Massive intestinal hemorrhage as the first sign of diverticular disease of the colon]. AB - Retrospective and prospective analysis of massive enterorrhagia (m.e.). The aim of the study is the analysis of frequent causes of m.e--diverticular disease of the colon (DDC). The total number of patients with m.e. was 154 and the proportion of DDC in this total group was 17%, i.m. 24 patients. The total number of patients with the acute symptomatic DDC was 198 and the proportion of haemorrhagic DDC in this total group was 13.3%. The dominant form is conservative therapy (88.4%), surgical therapy was performed in 11.6% of cases. Discontinuous types of operations predominated. The analysis of the group and comparison with problems of m.e. in the literature. PMID- 11392049 TI - [The acute phase reaction in laparoscopic and open surgery of inguinal hernias]. AB - The acute phase response to tissue injury is art of the wound healing process after surgery. The aim of study was to determine levels of acute phase proteins and levels of thrombocytes in patients with laparoscopic surgery (intraabdominal preperitoneal repair) and in patients with open surgery (tension free repair). Exclusion criteria in both groups of patients: malignity, diabetes mellitus, obesity (BMI > 30), infection, hypoproteinemia, hepatic or renal insufficiency and hypertension. Type of anaesthesia: general. Perioperative preventive antithrombotic medication: LMWH 5 days after surgery. The observed parameters were estimated before, one hour, 2nd and 7th days after surgery. Statistical test: ANOVA, statistical by significant difference p < 0.05. The results of the study demonstrate an increase of acute phase proteins CRP, OROSO and Fb in both groups of patients in comparison to their levels before surgery. In this respect we did not find a difference between the two types of operation. In patients with laparoscopic surgery the observed peak of FBG increase (+69%) was on the 2nd day after surgery followed by a slight drop of values in comparison to the results of open surgery patients with a FBG increase on the 2nd day (+42%) and with continuation on the 7th (%) postoperative day. The peak of CRP values was on the 2nd day in both groups. OROSO values increased even on the 7th day. The same situation occurred with Plt levels (p < 0.05). We suggested, that laparoscopic and open surgery of inguinal hernia repair are both followed by an acute phase response related to the tissue injury and this response perists even 1 week after surgery. But the recovery time of some parameters of the acute phase response (e.g. orosomucoid and fibrinogen levels) to the basical preoperative state is longer in patients with open type of surgery. We do not confirm differences in the degree of risk of postoperative thrombophilia in both types of surgery and suggest, that the prevention of thromboembolic complications is indicated in both types of surgery. PMID- 11392050 TI - [Gastrointestinal stromal tumor]. AB - The author draws attention to rare tumours of the digestive tract, their symptoms, treatment and prognosis. PMID- 11392051 TI - [Intestinal duplication. A rare disease in the differential diagnosis of cystic abdominal tumors]. AB - Intestinal duplications are described as uncommon congenital anomalies rarely diagnosed in adults. A case of young woman with obscure abdominal symptomatology is demonstrated. After investigation the cause of it was disclosed surgically. During operation a cystic ileal duplication was exposed and resected. Despite of difficulties calling for another operation, the case was successfully solved. Based on the author's experience some aspects of these diseases--etiopatogenesis, symptoms, possibilities of examination and complications are discussed and surgery as the main treatment is emphasized. PMID- 11392052 TI - [Treatment of the agitated patient in surgical practice]. AB - The authors discuss in their paper treatment of amented delirious conditions as a complication after an urgent or elective surgical operation. Contrary to hitherto applied practice, where treatment is started only at the onset of the mental disorder, the authors administered treatment in selected patients immediately after surgery. The results revealed a marked decrease of amented delirious conditions in the group of patients where treatment was started immediately after surgery. PMID- 11392053 TI - [Non-penetrating thoracic injuries]. AB - The authors compare the causes and severity of non-penetrating chest injuries during two three-year periods: 1986-1989 and 1996-1999. Within the 10-year interval the incidence of blunt chest injuries increased by almost 124%, the mean age of the casualties reduce by five years and the most frequent cause were road accidents. Chest injuries are in more than half the casualties part of multiple injuries or polytraumatism. During the period 1996-1999 the authors recorded more serious intrathoracic injuries, incl. severing of the v. azygos in three patients. With regard to the danger associated with delay it is essential that these casualties should be taken from the site of the accident directly to a specialized department. PMID- 11392054 TI - Vasoconstrictive drugs increase carbonic anhydrase I in vascular smooth muscle while vasodilating drugs reduce the activity of this isozyme by a direct mechanism of action. AB - Carbonic anhydrase (CA) is a zinc enzyme that catalyses the reversible hydration reaction of CO2 and plays a major role in the acid-base balance. We have previously shown that certain vasoconstrictive therapeutic agents increase CA I activity whereas vasodilating drugs reduce the activity of this isozyme by a direct mechanism of action. In this paper we studied the effect of other vasoconstrictive and vasodilating agents on CA I activity in order to elucidate the involvement of vascular smooth muscle CA I in vasoconstrictive and vasodilating processes. We studied the in vitro effects of noradrenaline, prostaglandin F2 alpha, thromboxane A2, leukotriene B4, angiotensin II, vasopressin, indomethacin, prazosin, hydralazine, clonidine, reserpine, prostaglandin I2, indapamide, furosemide, amlodipine, verapamil and irbesartan on purified human red blood cell CA I and vascular smooth muscle CA I isolated from rabbits. In vivo, we selected six groups of five rabbits each, which were administered the following substances in acute experiments: orciprenaline (group 1), desmopressin (group 2), verapamil (group 3), irbesartan (group 4), acetazolamide (group 5) and placebo (control group). Vascular smooth muscle CA I activity and systolic blood pressure were determined and compared with those of the control group. In vitro results showed that all the vasoconstrictive agents studied increased purified and human erythrocyte CA I activity as well as vascular smooth muscle CA I, while vasodilating substances reduced the activity of isozyme by a direct mechanism of action. The same results obtained in vivo showed that activation of vascular smooth muscle CA I increased blood pressure while its inhibition reduced blood pressure. The results of this study suggest that pHi changes, induced by activating or inhibiting CA I in vascular smooth muscle, might be responsible for changes in vascular tonus. PMID- 11392055 TI - Intracerebroventricular capsaicin and food intake in the rat. AB - To investigate whether capsaicin affected food intake and body weight and/or interfered with the mechanisms regulating the body temperature, capsaicin was administered intracerebroventricularly to rats at a daily dose of 25 micrograms of 5 microliters of vehicle for 10 consecutive days. The vehicle consisted of 5% ethanol plus 5% Tween 80 in isotonic saline. The ambient temperature (Ta) was 22 degrees C. Two control groups were used: the first group was treated with the vehicle but its concentration was lowered 10 times in order to avoid the wellknown brain toxicity of Tween 80; the second group was injected with NaCl 0.9% alone. The subsequent observation period lasted 38 days and the rats were exposed to a Ta of 22, 32, 35, 10 and 22 degrees C for different times. Food intake and body weight showed a remarkable decrease during the treatment period in both the capsaicin- and vehicle-treated groups. During the observation period, food intake behavior and body weight evolution were different among the three groups of treated rats. The influence of Ta was most apparent in the capsaicin treated rats that showed a long-lasting deficit in their body weight and a clear inability to regulate their body temperature (Tb) in a warm environment (Ta 35 degrees C). However, the capsaicin-treated rats began to recover from the weight loss when they were finally housed at Ta 22 degrees C. Tween 80-treated rats began to recover from the weight loss earlier, the pretreatment body weight having been reached during Ta 32 degrees C exposure, i.e., 12 days after the end of the treatment. Since capsaicin did not reduce food intake for a long time during the recovery process, the effect of capsaicin on Ta and body weight seems to be related to metabolic processes. Under the conditions of the present study, neither capsaicin nor Tween 80 seem to have permanently impaired the animals' energy balance regulation. PMID- 11392056 TI - Effects of topical diclofenac (DHEP plaster) on skin, subcutis and muscle pain thresholds in subjects without spontaneous pain. AB - The aim of this study was to determine whether topical application of diclofenac hydroxyethylpyrrolidine (DHEP) modifies somatic pain sensitivity in subjects without spontaneous pain. Twenty male subjects (aged 19-40 years), who had not reported any pain for at least 1 month, underwent measurement of pain thresholds to bilateral electrical stimulation in the quadriceps muscle and overlying subcutis and skin. Following the double-blind study design, one diclofenac adhesive plaster (13 x 10 cm; 180 mg DHEP) was then applied over one quadriceps while a matched placebo plaster was placed contralaterally. Each subject was given two other plasters (diclofenac and placebo) and instructed to substitute those over the quadriceps after 12 h and to wear them for a further 12 h. Thirty minutes after removal of the second plasters, thresholds. were remeasured in all subjects as on the previous day. Thresholds at the first evaluation were within normal range in nine subjects (group 1) and below normal in muscle (hyperalgesia) in the remaining 11 (group 2). No significant changes were observed in skin or subcutis thresholds after diclofenac or placebo treatment in either group. In contrast, muscle thresholds significantly increased after diclofenac compared with placebo treatment (group 1: p < 0.05; group 2: p < 0.007); the increase was significantly higher in group 2 than in group 1 (p < 0.002). Topical application of diclofenac had a selective hypoalgesic effect on muscles, which was more pronounced in the case of hyperalgesia. These results suggest that the preparation is particularly effective in the treatment of algogenic conditions of deep somatic tissues. PMID- 11392057 TI - One-month follow-up of patients treated by intravenous clodronate for acute pain induced by osteoporotic vertebral fracture. AB - Disodium-clodronate is one of the most experimented drugs for the management of bone pain caused by vertebral fractures induced by skeletal metastases and myeloma as well as by osteoporosis. In a previous study, treatment with intravenous disodium-clodronate 300 mg/day in saline produced satisfactory relief of moderate or severe back pain in 15 patients with painful vertebral crush. The present study examines the general behavior of pain in patients suffering from vertebral crush when treated with intravenous disodium-clodronate and after discontinuation of medication. At the end of treatment, pain relief was significantly greater in patients treated with disodium-clodronate. After discontinuation of treatment the patients who had been taking disodium-clodronate for pain due to vertebral crush were generally better than those previously on acetaminophen. At the end of follow-up, no significant differences were found between the two groups. In our study, the use of disodium-clodronate was associated with a reduction in the number of days the patients experienced severe pain, which suggests that disodium-clodronate is a first-line drug in the treatment of recent vertebral crush. PMID- 11392058 TI - Comparative effect of theophylline and aminophylline on theophylline blood concentrations and peripheral blood eosinophils in patients with asthma. AB - The comparative effects of a new theophylline preparation (Theodrip) and aminophylline on blood concentrations of theophylline were examined in 74 patients with asthma. Subjects were intravenously administered 200 mg of Theodrip or 250 mg of aminophylline for 1 h. The mean increases in blood theophylline concentration after Theodrip or aminophylline administration were 8.80 +/- 1.80 mg/l and 8.81 +/- 2.15 mg/l, respectively. In addition, these patients were divided into four groups based on baseline theophylline concentrations before infusion of Theodrip or aminophylline: i) naive patients (not administered theophylline); ii) those with a baseline theophylline concentration of 0-5 mg/l; iii) those with a baseline theophylline concentration of 5-10 mg/l; iv) those with a baseline theophylline concentration of 10-15 mg/l. Mean increases in blood theophylline concentration after administration of Theodrip in each group were similar to those after aminophylline administration. We found no significant differences between Theodrip and aminophylline. However, when the comparative effects of Theodrip and aminophylline on peripheral blood eosinophil counts were examined, Theodrip, but not aminophylline, reduced blood eosinophil counts. With acute exacerbations of bronchial asthma, it is expected that Theodrip, but not aminophylline, may have an antiinflammatory effect. In conclusion, it is suggested that Theodrip is a more useful drug than aminophylline in patients with acute exacerbations of bronchial asthma. PMID- 11392059 TI - The effect of an Na+/H+ exchange inhibitor, SM-20550, on ischemia/reperfusion induced endothelial dysfunction in isolated perfused rat hearts. AB - The aim of this study was to test the protective effect of an Na+/H+ exchange (NHE) inhibitor, SM-20550, on ischemia/reperfusion-induced endothelial dysfunction. Isolated rat hearts were subjected to 30 min of global ischemia followed by 20 min of reperfusion and their responses to the endothelial dependent vasodilator, acetylcholine, and the endothelial-independent vasodilator, nitroglycerin, before and after ischemia were examined. Acetylcholine-induced relaxation was impaired after ischemia/reperfusion while nitroglycerin induced relaxation was not. Administration of 1-10 nmol/l SM-20550 [N-(aminoiminomethyl)-1,4-dimethyl-1H-indole-2-carboxamide methanesulfonic acid] before and after ischemia prevented impairment of acetylcholine-induced relaxation. To further understand the mechanism of SM-20550 in protecting endothelial function, we measured the inhibitory activity of SM-20550 on NHE in cultured endothelial cells. SM-20550 (1-100 nmol/l) inhibited recovery from acidosis induced by an NH4Cl prepulse in a concentration-dependent manner. Oxygen radicals from endothelial cells and leukocytes are one of the major sources of endothelial cell injury during ischemia and reperfusion. Consequently, we tested the effect of SM-20550 on H2O2-induced endothelial cell injury. SM-20550 (100 1,000 nmol/l) prevented H2O2-induced cell injury measured by lactate dehydrogenase assay. In conclusion, SM-20550 inhibited NHE in endothelial cells, protected ischemia/reperfusion-induced endothelial dysfunction and prevented H2O2 induced endothelial cell injury at higher concentrations. PMID- 11392060 TI - Immunomodulating effects of extracorporeal photochemotherapy in rat experimental allergic encephalomyelitis. AB - Experimental allergic encephalomyelitis (EAE) is a well-established model of human multiple sclerosis that is commonly used to evaluate the possible effectiveness of new treatments in this disease. Extracorporeal photochemotherapy (ECP) is an immunomodulating procedure currently used in several non-neurological diseases that, like multiple sclerosis, are likely to be due to T-cell-mediated autoimmunity. In this study we examined the effect of ECP using the EAE paradigm in the Lewis rat. In our model, ECP induced a significant modulation in peripheral blood T-cell distribution, changes which are typical of EAE. Remarkably, this effect was closely correlated with the clinical and pathological results, which showed reduced severity of the disease in the ECP-treated EAE animals vs. the EAE alone rats. We conclude that ECP induces modifications in the immunological events that occur during the course of EAE in rats, thus giving support to the hypothesis that it could be used in the treatment of multiple sclerosis. PMID- 11392061 TI - Effect of intramuscular clodronate on bone mass and metabolism in osteoporotic women. AB - Bisphosphonates have significantly improved the treatment of postmenopausal osteoporosis. However, when administered orally, gastric intolerance is their main adverse effect. This makes patients reluctant to undergo treatment. As an alternative, intramuscular (i.m.) administration may be more acceptable to patients undergoing long-term treatment. Since clodronate has been easily available in Italy for many years, we carried out this study to evaluate its effect on bone mineral density (BMD) in 36 osteoporotic postmenopausal women who were intolerant to oral administration of bisphosphonates. Patients received 100 mg of clodronate i.m. every 10 days together with 500 mg/day of calcium orally for 2 years. A control group of 32 women received only oral calcium at the same dose and for the same length of time. In the control group a progressive but not statistically significant decrease in BMD was observed in the spine and femoral neck over the 2-year follow-up. In contrast, patients treated with clodronate had a statistically significant increase in BMD in the spine at the first yearly check-up (+2.63%) and a further but not statistically significant increase during the second year of treatment (+0.59%). The increase in BMD at the femoral neck was not statistically significant during the first and second years of treatment, being 1.21% and 0.37% respectively. In the women treated with clodronate, hydroxyproline was significantly suppressed. All the patients reported local pain at the injection site. This led 11.11% of the subjects to withdraw from treatment. PMID- 11392062 TI - Three-month follow-up of shoulder-hand syndrome induced by phenobarbital and treated with gabapentin. AB - In the present study we evaluated the 3-month follow-up of 14 subjects with phenobarbital-induced shoulder-hand syndrome after discontinuation of their previous pharmacological treatment (group 1: gabapentin 100 mg/day; group 2: acetaminophen 3 g/day for 3 months). The aim of this study was to evaluate pain and joint function in each group after cessation of treatment and to compare the results in the two groups. The result for pain and joint function was better in the seven patients previously treated with gabapentin. PMID- 11392063 TI - Interaction between monensin and CCl4 after chronic treatment: dolichol and retinol content of rat liver sinusoidal cells. AB - The aim of this investigation was to study retinol, which is known to decrease in hepatic stellate cells during fibrogenesis, and dolichol, which influences membrane fluidity and decreases in liver injury, in freshly isolated liver parenchymal and nonparenchymal cells after intoxication of rats with CCl4 combined with the ionophore monensin for 3, 5 and 7 weeks. To study the interrelationship between dolichol and vitamin A transport, a load of vitamin A was given to batches of rats 3 days before sacrifice. Monensin did not modify the action of CCl4 in hepatocytes. On administration of CCl4 and CCl4-monensin, dolichol decreased independently of vitamin A load, while retinol increased, especially when a load of vitamin A was given to rats 3 days before sacrifice. Hepatocytes appeared to no longer be able to export or metabolize vitamin A. In a subfraction of hepatic stellate cells (Ito-1 cells) dolichol always decreased, while retinol was no longer stored after each treatment; dolichol and retinol showed the same behavior but the decrease was more pronounced in monensin after vitamin A load and after 3 weeks. These data support the hypothesis that by modulating membrane characteristics, dolichol might be involved in intracellular or intercellular retinol transport and that altered transport between hepatocytes and Ito-1 cells might accompany liver injury. The data regarding another subfraction, Ito-2 cells, partly resemble those for the Ito-1 fraction and are in agreement with the heterogeneity of hepatic stellate cells. In Kupffer and sinusoidal endothelial cells, dolichol and retinol content was not homogeneous and was only slightly altered after the treatments. Monensin and CCl4 are not interactive. Although both drugs alter membrane lipids, their association allows some sinusoidal cell responses to be differentiated. PMID- 11392064 TI - Comparative effects of diltiazem with enflurane or desflurane on myocardial contractility and heart rate in the isolated rat hearts. AB - BACKGROUND: The volatile anesthetics may reduce cardiac contractility by limiting both membrane Ca2+ entry and altering intracellular Ca2+ release. Additional pharmacological effects of calcium channel blockers could potentially enhance anesthetic-induced depression. The aim of this study was to compare the direct cardiac effects of enflurane and a new volatile anesthetic, desflurane, in combination with diltiazem on the isolated Sprague-Dawley rat heart. METHODS: After stabilization period isolated rat hearts (n = 40) were perfused with an oxygenated modified Krebs' solution at 55 mmHg equilibrated with 1, 2 and 3 MAC of enflurane (1.7, 3.4 and 5.1 vol% respectively) or desflurane (6, 12 and 18 vol% respectively) in combination with 100 ng/mL diltiazem at 36 degrees C. Isovolumetric left ventricular pressure (LVP), rate of change of ventricular pressure (dp/dt), spontaneous heart rate and coronary flow were measured. To examine the indirect metabolic effect due to autoregulation of coronary flow, O2 delivery (DO2), myocardial O2 consumption (MVO2) and percent O2 extraction (POE) were also monitored. RESULTS: Diltiazem plus enflurane or desflurane depressed LVP and dp/dt dose-dependently. Enflurane plus diltiazem significantly decreased heart rate more than desflurane plus diltiazem in a dose-dependent manner. Desflurane plus diltiazem significantly increased coronary flow more than enflurane plus diltiazem and oxygen delivery increased proportionally with coronary flow. But there were statistically insignificant dose-dependent increases in both groups. Myocardial oxygen consumption and percentage of oxygen extraction were also decreased dose-dependently in both groups. Bradydysrhythmia that accompanied atrioventricular dissociation occurred with diltiazem plus high enflurane or desflurane concentration at an incidence of 46% and 40% respectively. CONCLUSIONS: These in vitro results demonstrate that diltiazem plus enflurane or desflurane depresses left ventricular contractile function and diltiazem plus enflurane causes higher incidence of bradydysrhythmia more than equivalent levels of diltiazem plus desflurane. PMID- 11392065 TI - Study of propofol in bovine aortic endothelium: I. Inhibitory effect on bradykinin-induced intracellular calcium immobilization. AB - BACKGROUND: Propofol has been found to affect the intracellular calcium concentration with clinical manifestations of hypotension and bradycardia. The purpose of this study is to examine the effect of propofol on intracellular calcium immobilization in bovine aortic endothelium under the stimulation of bradykinin. METHODS: In order to validate the effect of propofol on the alteration of intracellular calcium concentration, we used the cultured bovine endothelial cells (Gm 7372a) to measure the calcium immobilization within the cells preincubated with or without propofol of clinical concentration. Using Fluo 3 staining and a fluorescence spectrophotometer (confocal microscope), intracellular calcium immobilization was demonstrated by the appearance of "hot spots" within the cytoplasm and perinuclear regions after addition of bradykinin to the cells. The changes of fluorescence density measured within these areas versus the effect of time were analyzed and compared with the cells in control group. RESULTS: After addition of bradykinin, intracellular calcium hot spots increased dramatically within seconds and reached a maximal level within 20 seconds. The concentrations of calcium gradually decreased to a constant level after about 3 min following the addition of bradykinin to the cells. With pretreatment of propofol at 0.01 mM and 37 degrees C for 30 min, the immobilization of intracellular calcium from the intracellular stores were significantly inhibited that was demonstrated by the decreased appearance of hot spots when compared with control. CONCLUSIONS: Our data demonstrated that under the stimulation of bradykinin, propofol at 0.01 mM, could inhibit intracellular calcium release from the intracellular stores in bovine aortic endothelial cells. This phenomenon might explain the possible mechanism for the clinical manifestations of hypotension and/or bradycardia associated with propofol. PMID- 11392066 TI - Absence of the preemptive analgesic effect of dextromethorphan in total knee replacement under epidural anesthesia. AB - BACKGROUND: Previous studies have shown that dextromethorphan (DM), a N-methyl-D aspartate (NMDA) receptor antagonist, produces a preemptive analgesic effect on post-operative pain. The aim of this study was to further examine the preemptive analgesic effect of intramuscular (i.m.) DM injection on unilateral total knee replacement (TKR). METHODS: Sixty-four ASA I-III patients scheduled for unilateral TKR surgery were randomly allocated into three groups in a prospective double-blind manner. All patients received epidural anesthesia without any premedication. An initial bolus dose of 2% lidocaine (15-20 mL) followed by a maintenance dose of 8-10 mL/h was decided. Fentanyl (1.5 micrograms/kg) and diazepam (2 mg) were given i.v. before epidural catheter insertion. The epidural catheter was placed via the L2-L3 or L3-L4 interspace and advanced for 5 cm cephalad [corrected]. Patients received i.m. injection of 20 mg chlorpheniramine (CPM) before surgery as control (group C, n = 22). For the study groups, patients were given an i.m. injection containing 40 mg DM and 20 mg CPM, before (group B, n = 22) or after surgery (group A, n = 20), respectively. Postoperation, patients received intravenous morphine by means of a patient controlled analgesia (PCA) device for pain relief. The time to the first pull of PCA trigger, morphine consumption, worse pain scores (resting and incidental), and analgesics related side effects were recorded at 1, 2, 4, 8, 24, 48 and 72 h after surgery. RESULTS: The time from the end of operation to the first PCA trigger were 31.2 +/- 5.2 min in group C, 67.3 +/- 11.1 min in group B (P < 0.05, compared with group C) and 61.8 +/- 7.2 min in group A (P < 0.05, compared with group C) respectively. The relevant pain score at resting, observed at the 8 h postoperatively was respectively 4.2 +/- 0.1 in group C, 3.7 +/- 0.2 in group B (P < 0.05, compared with group C) and 3.4 +/- 0.2 in group A (P < 0.05, compared with group C); and at the 24 h was 3.1 +/- 0.2 in group C, 2.4 +/- 0.2 in group B (P < 0.05, compared with group C) and 2.5 +/- 0.1 in group A (P < 0.05, compared with group C) respectively. There were no significant differences in actual morphine delivery and frequency of PCA triggering at all time among the three groups. Moreover, there was also no significant statistic difference in morphine associated side effects among the three groups. CONCLUSIONS: In the present study, we failed to observe any preemptive analgesic effect of DM (40 mg, i.m.) on postoperative pain in patients who received TKR under epidural anesthesia, however, DM given either before or after surgery augmented other analgesic (morphine) to offer a better pain relief. PMID- 11392067 TI - Thoracic epidural analgesia with morphine does not prevent postthoracotomy pain syndrome: a survey of 159 patients. AB - BACKGROUND: This retrospective study sought to determine the incidence of postthoracotomy pain syndrome (PTPS), and whether epidural morphine for the postoperative analgesia could prevent the development of PTPS. METHODS: We reviewed the charts of 372 patients who had undergone thoracotomy. The majority underwent general anesthesia (GA) combined with thoracic epidural analgesia (TEA). Of the 372 patients, only 159 (42%) were available for interview. Patients were divided into two groups based on the duration of pain, i.e., pain group (pain > 3 months, n = 65) and pain-free group (pain < 3 months, n = 94). RESULTS: Both groups were comparable regarding sex, age, weight, height, smoking, alcohol ingestion, education, marital status, duration of surgery, and the number of patients either receiving GA plus TEA or GA alone. About 41% of the patients experienced PTPS that persisted for 21 +/- 12 mon (follow-up: 28 +/- 12 mon). Most pain was mild or moderate and was usually described as being only a discomfort. Only 6.2% suffered severe pain with shooting, aching, burning or numbness. Patients with PTPS suffered more depression and insomnia. The incidence of PTPS was not different in patients who received GA alone or GA plus TEA (39% vs. 42%). CONCLUSIONS: Epidural morphine for postoperative analgesia that continued for 3 days appeared to have no effect in the prevention of PTPS. PMID- 11392068 TI - Failure of prevention against postoperative vomiting by ondansetron or prochlorperazine in patients undergoing gynecological laparoscopy. AB - BACKGROUND: Ondansetron has been approved for the treatment and prevention of postoperative emesis. Since it is presumably considered to possess potent antiemetic effect with fewer side effects, the administration of ondansetron to inhibit emesis in patients following gynecological laparoscopic surgery might be recommendable. Hence, we examined the effects of intravenous ondansetron at dosage of 4 and 8 mg in comparison with intravenous prochlorperazine at 5 mg and placebo. METHODS: A total of 120 patients were allocated randomly into 3 groups. Group 1 patients who served as control were given NaCl 0.9% 4 mL (placebo) intravenously (i.v.); patients in group 2 and group 3 were given ondansetron 4 mg ondansetron 8 mg i.v. respectively; patients in group 4 were given prochlorperazine 5 mg i.v. Premedication was omitted. RESULTS: Logistic regression analysis adjusted for prognostic factors revealed no significant difference between 5 mg prochlorperazine group and 4 mg or 8 mg ondansetron group as compared over the 24 h study period. CONCLUSIONS: The results of this study suggest that i.v. 4 or 8 mg ondansetron and 5 mg prochlorperazine were not effective in prevention of postoperative emesis in patients undergoing gynecological laparoscopy. Since the cost of ondansetron is high, its routine use for prevention against postoperative nausea and vomiting is not be recommended clinically because of its uncertain benefit. PMID- 11392070 TI - Coronary artery bypass grafting on the beating heart using the octopus method--a case report. AB - A 74-year-old male was admitted due to chest tightness for one month. He had received percutaneous transilluminal coronary angioplasty (PTCA) because of single-vessel disease one year ago. Cardiac catheterization examination carried out during this admission revealed 90% stenosis of the left anterior descending coronary artery (LAD) and circumflex branch (CX). Because another attempt of PTCA was considered not optimal, the patient was advised to undergo surgical grafting to which he consented. After expediently balancing the merits and demerits of every practicable surgical procedure--the essential determinant in selection of which was that the patient's condition and criteria of indication of that procedure were in perfect harmony--we decided to carry out minimally invasive direct coronary artery bypass (MIDCAB) with the application of Octopus tissue stabilizer. We report the surgical course and anesthetic management of the patient and discourse some detail in MIDCAB. PMID- 11392069 TI - Gene therapy: a novel method for the treatment of myocardial ischemia and reperfusion injury--mini-review. AB - Myocardial ischemia and reperfusion injury (MI/R) represents important sequelae of clinical events. Historically, a number of approaches including, surgical intervention, pharmacological therapy and physical exercise regimes have been prescribed for the treatment of patients with cardiovascular disease. Recently, however, attention has focused upon more novel approaches using gene-based therapies to treat cardiovascular and MI/R. This mini-review will examine the role that heat shock proteins (HSP), in particular the HSP70 family, and the antiapoptotic protein Bcl-2 play in myocardial protection. Also examined in this review are several techniques including adenovirus and Japan-Liposomal method for delivering genes into the myocardium. PMID- 11392071 TI - Rhabdomyolysis after a long-term thoracic surgery in right decubitus position. AB - We report a rare case who developed rhabdomyolysis associated with the use of the right decubitus position for 10 h during thoracotomy with lobectomy. It appears that an increasing of the compartment pressure may induce reperfusion injury of the ischemic muscle by prolonged compression of the gluteal and flank muscles against the operation table. Early recognition and aggressive treatment with intravenous fluid and diuresis may prevent the development of acute renal failure. Adequate prevention in high-risk patients, early diagnosis and aggressive treatment are the keys to a successful recovery. PMID- 11392072 TI - Prolonged paralysis associated with succinylcholine--a case report. AB - Prolonged paralysis is a rare complication of succinylcholine. We report a case of palmar hyperhidrosis who underwent transthoracic endoscopic sympathetic ganglionectomy under general anesthesia with a facemask developed prolonged paralysis following the operation. The prolonged paralysis supposedly caused by intraoperative use of succinylcholine was recognized and confirmed by laboratory examination which showed low pseudocholinesterase activity. In this report, we describe the course of the event and discuss the causes of delayed awakening associated with anesthesia and its management. PMID- 11392073 TI - [Qualitative and quantitative detection of hepatitis C virus RNA by PCR technique. Monitoring of viral copies after liver transplantation]. AB - The authors demonstrate the HCV nucleic acid amplification method is not wide spread in Hungary yet. The HCV-RNA is usually detectable 2-4 weeks after infection independently the immunostate of the patients. The authors help to select the adequate measurement(s) in logical order when HCV infection is suspected. The benefit of the PCR method is emphasized. Monitoring of the HCV-RNA titer of the liver transplanted patients promotes to establish the fluctuation of HCV-RNA copies and the effectivity of therapy following transplantation. The detection of HCV-RNA by PCR method is a proof of an acute or chronic infection and rules out past infection. The quantitative PCR measurement is useful for determination of indication and control of efficacy of antiviral therapy. PMID- 11392074 TI - [Effectiveness of high-frequency oscillatory ventilation in the treatment of lung hypoplasia in connection with intrauterine growth retardation and maternal pre eclampsia]. AB - The authors examine in a retrospective study the effectivity of high frequency oscillatory ventilation in group of intrauterine retardated babies with clinical signs of lung hypoplasia. During one year period they treated 12 intrauterine retarded patients with high frequency oscillatory ventilation because of failure of conventional ventilation. The indications were in seven cases the high pCO2 level and in five ones low saturation in spite of surfactant administration. The birth weight of babies were between 770 grams and 2150 grams, and theirs gestational age between 29 and 35 weeks. Four patients died. The causes of death were in two cases severe cardiomyopathy and at two patients intraventricular haemorrhage. Two survivors are suffering of bronchopulmonary dysplasia. They suggest that immediate high frequency ventilation reduces the period of mechanical ventilation and the number of residual picture. They suggest, that lung hypoplasia is an underestimated diagnosis in intrauterine retardated babies. If the cause of respiratory failure in cases complicated with maternal preeclampsia is not only hyaline membrane disease it is necessary to look for signs of lung hypoplasia and to follow the treatment with high frequency oscillatory ventilation. PMID- 11392075 TI - [The role of estrogen receptor, vitamin D receptor and calcium receptor genotypes in the pathogenesis of colorectal cancer]. AB - In this study, the Xbal polymorphisms of the estrogen-, the Bsml polymorphism of the vitamin D- as well as the A986S polymorphism of the calcium-sensing receptor genes were investigated in 56 patients with colorectal cancer. The expression of erbB-2, epidermal growth factor receptor, ras, p53 and their relationship to estrogen-, vitamin D- and calcium-sensing receptor genotypes were also studied. In subjects exhibiting XX genotype of the estrogen receptor gene or bb genotype of the vitamin D receptor gene, erbB-2 expression was significantly lower compared to those with xx, Xx or BB, Bb (6/56 and 11/56 vs. 31/56 and 26/56; p = 0.0043 and 0.041). The presence of the XX alleles of estrogen receptor gene significantly correlated with the overexpression of the epidermal growth factor receptor expression in tumors, whereas in xx and Xx genotypes, significantly higher expression was seen (7/56 vs. 30/56; p = 0.049). Analyzing the combinations of the two gene allelic variants, we have found XXbb genotype to be associated with a significantly lower erbB-2 expression, compared to other combinations (Xxbb, XxBb, XXBb) (2/7 vs. 7/7, 4/5, 4/5; p = 0.0011). Patients with AA calcium-sensing receptor genotype were in higher UICC stages at the time of discovery of their disease than those with AS genotype. The AA allelic variant of the calcium-sensing gene was more frequent among patients with colorectal cancer compared to controls (36/56 vs. 36/112; p = 0.0004). Our observations raise the possibility that estrogen-, and vitamin D receptor gene polymorphisms accompanied with variable oncogene expression might influence the pathogenic processes resulting in the development of colorectal cancer. The A986S polymorphism of calcium-sensing receptor might also be a prognostic marker of the disease. PMID- 11392076 TI - [Results of surgical treatment in hemifacial spasm--the role of MR-angiography in detecting microvascular compression]. AB - The authors evaluated the follow-up results of microvascular decompression (sec. Janetta) in 8 patients with hemifacial spasm (HFS). Indication was based on there dimensional time of flight magnetic resonance angiography with 0.5T Elscint Gyrex V Dix equipment. Contrast material was administered in every case and maximum intensity projection and thin slice reconstruction were performed in three standard directions. Vascular contact with the facial nerve in the entry zone was identified on the symptomatic side in 10 patients. No contact was detected in 2 cases. Microvascular decompression was performed in 8 cases. The surgical and neuroradiological findings were identical in every cases. Five patients were completely free of HFS immediately after surgery, and another 2 patients became free of HFS during the next few weeks. Only 1 patient had uncured symptoms. In conclusion, the authors suggest that microvascular decompression of the facial nerve may evolve as the method of choice if vascular contact is proved by 3D TOF MRA. PMID- 11392077 TI - [History of the Imperial and Royal Garrison Hospital in Pest from its foundation until after the War of Independence]. PMID- 11392078 TI - [In memory of principal physician Elemer Tamas (1918-1997)]. PMID- 11392079 TI - [Renal and ureteral calculi in abdominal diagnostics. 1926]. PMID- 11392080 TI - Fatal leptospirosis, Azores islands. PMID- 11392081 TI - Progress towards poliomyelitis and dracunculiasis eradication, Sudan, 2000. PMID- 11392082 TI - Women and men at sea: gender debate aboard the hospital ship Maine during the Boer War, 1899-1900. PMID- 11392083 TI - The Jewish problem in U.S. medical education, 1920-1955. PMID- 11392084 TI - The history of bioethics: an essay review. PMID- 11392085 TI - [Functional morphology of the hypophyseal-adrenal-gonad system in different variants of prostatic hyperplasia]. AB - Two forms of prostatic hyperplasia are distinguished: fibro-glandular (FGH) and glandular-fibrous (GFH). An increase in the number of adenohypophysis cells producing gonadotrophic hormone, hyperplasia of the adrenal reticular zone, hyperplasia and hypertrophy of Leydig and Sertoli cells in the testis were observed in FGH. Signs of enhanced functional activity were observed in all cases of hyperplasia. In GFH there was an increase in the number of adenohypophyseal cells producing adrenocorticotrophic hormone, hyperplasia of the fascicular layer in the adrenals, atrophy of Leydig cells in the presence of Sertoli cell hyperplasia. The variant of prostatic hyperplasia is determined by the hormonal changes and this may help in choosing of approaches to therapeutic correction of prostatic hyperplasia and its early diagnosis. PMID- 11392086 TI - [Morphological characteristics of chronic hepatopathy in combination of HCV and chronic alcoholic intoxication]. AB - Biopsies from 35 patients and autopsy material from 20 patients who died of chronic hepatopathy were studied. Hydropic dystrophy and inflammation were predominant in HCV infection. Chronic alcoholic intoxication (CAI) was characterized by diffuse large-droplet fat dystrophy and liver fibrosis. In combination of HCV-infection with CAI sclerotic processes were pronounced and therefore liver cirrhosis is more frequent than in other groups. PMID- 11392087 TI - [Mechanisms of development of diabetic macroangiopathy]. AB - Pieces of the aorta and heart coronary arteries taken at autopsy of 100 patients who had died of insulin-dependent and non-insulin-dependent diabetes mellitus were studied histologically and immunohistologically. Atherosclerosis was more pronounced in diabetes mellitus patients than in patients with normal glycemic indices. Atherosclerosis was most severe in non-insulin-dependent diabetes. Diabetic microangiopathy of the vasa vasorum of large arteries promotes development of atherosclerosis. PMID- 11392088 TI - [Pathohistological diagnosis of precancerous conditions and cancer of the breast]. AB - It is suggested to improve histopathological diagnosis on the basis of mean ploidy of cell clones characterising separate stages of carcinogenesis. Additional ploidometric analysis of sections stained by Felgen enables differential diagnosis of benign (hyperplastic lesions and mild neoplasia) with malignant stages of carcinogenesis (grave intraepithelial neoplasia, carcinomas) in the breast. PMID- 11392089 TI - [Sudden arythmogenic death in hospitals after heart surgery]. AB - 29 cases of sudden heart death (SHD) which occurred in hospital after heart surgery within 1989-1998 are reported. More than 50% were SHD after aortocoronary shunting, it occurred in stenosing atherosclerosis of more than 3 arteries. An important role in tanatogenesis belongs to ischemic heart disease (IHD) with complications (chronic aneurysm of the left ventricle, postinfarction failure of the mitral valve) and combination of IHD with aortal heart deficiency which is followed by pronounced hypertrophy of the left ventricle which is an important factor of SHD risk. Interventricular defects and Fallot's tetrad are most frequent among heart malformations with SHD after heart surgery. Cardiomegaly, myocardial fibrosis, dilatation of the heart cavities, fibroelastosis of ventricular endocardium, anomalous chordal arrangement in the left ventricle, right ventricular lipomatosis are main factors of SHD of the arrhythmogenic type after heart surgery. Introduction of the new notion "sudden heart death at the hospital stage after heart surgery" is suggested. PMID- 11392090 TI - [Morphofunctional analysis of right ventricle cardiomyocytes in patients with Fallot's tetrad during first year of life]. AB - Material of intrasurgical biopsies of the right ventricular (RV) myocardium of 28 patients with Fallot's tetrad was studied with light and electron microscopy. RV cardiomyocytes were hypertrophic and showed signs of growth and differentiation. Myofibrillogenesis intensity directly correlated with the pressure gradient between RV and pulmonary artery. At the same time there was a structural alteration of mitochondria and formation of a specialized population of intermyofibrillar mitochondria. Continuing myofibrillogenesis in cardiomyocytes may be one of the causes of decreased myocardial contractility. PMID- 11392091 TI - [Syndrome of enlarged thymus gland in children with congenital heart defects]. AB - Material of the study represents samples of the enlarged thymus of 40 children aged from 1 month to 3.5 years obtained during surgery on the open heart. A thymus enlargement in the majority of children with heart malformations was due to its reactive hyperplasia similar to the first stage of acute (accidental) involution or changes observed in the thymus in the "rebound" phenomenon. In some cases B-lymphocytic hyperplasia was observed which is considered as a sign of the autoimmune disturbances. PMID- 11392092 TI - [Microscopic polyarteritis in a patient with hypertension]. AB - A case of microscopic polyarteritis in a male aged 45 years having hypertension for 22 years is reported. There were clinical manifestations (hemorrhages, subfebrile temperature, arthralgias and others) and damages in many organs: productive-necrotic alveolitis, capillaritis of various organs, morphological features of hypertension, atherosclerosis, subendocardial myocardial infarction. The leading clinical feature was rapid progression of renal and pulmonary-heart failure. PMID- 11392093 TI - [Pathological changes in spiral arteries in systemic lupus erythematosus]. AB - The uterus removed at 20-week pregnancy in a female with grave lupus erythematosus complicated with an antenatal fetus death was studied. Signs of arthrosis typical for late pregnancy were found in the spiral arteries of basal decidua in the zone of contact between mother tissue and trophoblast. Fetal death was due to massive thrombosis. PMID- 11392094 TI - [Combination of generalized intestinal candidiasis and balantidiasis in a child with Down syndrome]. AB - The above combination is probably due to immunodeficiency characteristic of Down's syndrome. PMID- 11392095 TI - [Morphological characteristics of chordoma of the temporal bone: light amd electron microscopy]. AB - For the first time chordoma of the temporal bone in a female aged 24 years is described. The tumor was studied histologically and ultrastructurally. PMID- 11392096 TI - [Classification and coding of clinical-anatomical forms of tuberculosis]. AB - Principal forms of tuberculosis are morphologically characterized as well as their complications. The comparison with ICD-10 is made. PMID- 11392097 TI - [Pathogenesis of chronic hepatitis B]. AB - The following problems are considered in chronic HBV infection: viral replication within and outside the liver, heterogenicity of genotypes and mutations of viral genome, immunogenetic basis of the host, direct cytopathic effect of the virus and immunological reactions induced by the virus. PMID- 11392098 TI - [Insulin-like growth factors in lung precancer]. AB - Investigation of IGF-system resulted in distinguishing two groups of processes depending on the size of the bronchi. Goblet cell metaplasia, basal cell hyperplasia with or without atypical features, squamous cell metaplasia, dysplasia 1-3, neuroendocrine cell hyperplasia, epithelial atrophy were found in the large bronchi. The same changes occurred in the bronchioli and alveoli plus adenomatous hyperplasia with or without atypia, oval and narrow structures with epithelial atrophy, atypia and proliferation. Common property was accumulation in the cells of various members of IGF system: high accumulation of IGF II, IGFBP 1,2,3,4 in dysplasia; the same plus IGFBP 11,2,3,4,5 in squamous metaplasia. Tuberculous granuloma cells are an important source of IGF II, IGFBP 2,3,4,5,6 playing a role in proliferation and apoptosis of the pulmonary epithelium. PMID- 11392099 TI - The effect of fluvastatin on fibrinolytic factors in patients with hypercholesterolemia. AB - Several studies have shown cardiovascular benefit in treating hypercholesterolemia with HMG-CoA reductase inhibitor. However, in addition to the lowering of cholesterol, the beneficial effects of this inhibitor reflect other pharmacological activities. Whether these beneficial effects are partly mediated by changes in fibrinolytic factors remains to be proven, since clinical studies on the effects of HMG-CoA reductase inhibitors on fibrinolytic factors have not yielded consistent results. The purpose of this study was to evaluate the effects of fluvastatin on fibrinolytic factors in hypercholesterolemic patients. After 6 weeks on a low-fat, low-cholesterol diet, 23 outpatients known to have primary hypercholesterolemia with low density lipoprotein cholesterol (LDL-C) > or = 130 mg/dl with at least 2 risk factors or fasting LDL-C > or = 160 mg/dl were selected for the study. Venous blood samples were collected at baseline and at 8 weeks after fluvastatin therapy (40 mg/day) to measure of tissue plasminogen activator (t-PA), plasminogen activators inhibitor-1 (PAI-1), fibrinogen, D-dimer and lipid profile. After 8 weeks of therapy, fluvastatin reduced serum cholesterol by 11% (261.9 mg/dl vs 233.2 mg/dl, P < 0.01) and LDL-C by 22% (191.9 mg/dl vs 149.3 mg/dl, P < 0.01). D-dimer was significantly decreased (0.38 ng/L vs 0.28 ng/L, P = 0.02) and tPA, PAI-1 and fibrinogen tended to decrease after therapy. Fluvastatin therapy improved fibrinolytic profile; the result of this study may in part explain the benefit of HMG-CoA reductase inhibitor on cardiovascular system other than lipid lowering. PMID- 11392101 TI - Relationship between outcome of proliferative vitreoretinopathy and results of tissue culture of excised preretinal membranes. AB - The purpose of this study was to explore the relationship between clinical post surgery outcome of proliferative vitreoretinopathy (PVR) and the laboratory results of tissue culture, the specimens of which were excised of pre- or subretinal membranes from PVR patients. Surgically excised membranes from 25 PVR patients were microdissected into small pieces and plated into culture dishes with F12 medium supplemented with 30% fetal bovine serum. After primary culture became confluent, cells were passaged in subculture with F12 medium (10% fetal bovine serum). PVR patients were followed-up after surgery for an average of 21 months. The clinical outcome was compared, according to the growth pattern of the cells derived from the explanted membranes. In 25 PVR patients, 16 cases showed cell migration in the membrane, and cells grew rapidly to confluence in the primary culture in 7 cases. All active growing cells were identified as retinal pigment epithelial (RPE) cells by immunocytochemistry. In 7 cases with active cell growth, all had recurrent retinal detachment. In 18 cases without active cell growth, only 4 cases had the same outcomes. Statistical study showed that the difference between these two groups was significant (P < 0.01). These results indicate that the growth capacity of cultured RPE derived from excised membranes of PVR patients strongly influenced the prognosis for surgery. PMID- 11392100 TI - Simplified 13C-urea breath test for the diagnosis of Helicobacter pylori infection--the availability of without fasting and without test meal. AB - The conventional 13C-urea breath test (13C-UBT) for detecting Helicobacter pylori infection was performed during fasting state and with a test meal to delay gastric emptying during the test. For the convenience of propagating this test, we assessed the availability of non-fast and without test meal in 13C-UBT for the diagnosis of H. pylori infection. One hundred and five consecutive patients who received endoscopic examination were studied. All of them received endoscopic biopsy for urease test, culture and histopathology to determine whether there was a presence of H. pylori infection. Each patient received four separate 13C-UBT under the following four testing conditions. Test I) fasting state with test meal (100 ml fresh milk), Test II) non-fast (taking usual food) but with test meal, Test III) fasting state without test meal, and Test IV) non-fast and without test meal. The excess delta 13CO2 values were calculated via the breathed samples that were collected at 15 minutes after ingestion of 13C-urea. There were 61 H. pylori positive and 44 negative patients in this study, with an excess delta 13CO2 values 3.0, 4.0, 3.5 and 4.0 as a cut-off value in the four tests respectively, according to the ROC curve. The results of test I, a conventional procedure, had a good correlation with the gold standard. The sensitivity and specificity were 100% and 95% respectively. The alternative procedures in other tests also have high sensitivity and specificity at 15-minute detecting time. The sensitivity of the tests II, III and IV at 15-minute detecting times were 98%, 98% and 100%, and the specificities of those were 95%, 98% and 95% respectively. We therefore suggest that fasting and test meal possibly be omitted when the cut-off value is 4.0 per mil in the simplified 13C-UBT (non-fast and without test meal, and detection at 15 minutes after ingestion of 13C-urea) which is a simple and available procedure for clinical diagnosis of H. pylori infection. PMID- 11392102 TI - Parents' psychosocial problems influencing the continuity of early intervention treatment in children with developmental delay. AB - Developmental delay is a chronic disorder, which has a significant and continuing impact on the developmental progress of children. Whether the developmentally delayed children have continuous early intervention treatment is important for the success of early intervention. Professionals in early intervention services have been encouraged to understand parents' psychosocial problems and to further enhance their understanding of care for their delayed children. The purposes of this study were two-fold. The first was to identify the parents' psychosocial problems. The second purpose was to find parents' psychosocial variables that could be used to significantly discriminate whether the retarded children have continuously received treatment programs. A questionnaire to investigate parents' psychosocial problems was implemented. It mainly consisted of twenty descriptive items concerning parents' psychosocial conditions. Parents used a four-point Likert scale to rate the likelihood of the items. Parents also responded as to whether their child had received treatments continuously. Subjects were the parents of children who were diagnosed to have developmental delay. Factor analysis and discriminant analysis was used for statistical data examination. Analyzed data were from 207 completely finished questionnaires. Six parents' psychosocial factors were extracted via factor analysis, and they accounted for 46.9% of the variance of the twenty items. The factors were identified as family income, family members' relationships, caring skills, supporting resources, acceptance of the retarded child, and self-psychological adaptation. About half of the children had received treatment continuously. Discriminant analysis revealed that family income was crucial to parents' capacity to meet the needs for treatment. PMID- 11392103 TI - The relationship between social support and well-being of rural elderly women. AB - The purpose of this study is to examine the relationship between social support and well-being of rural elderly women. A sample of 284 women aged 60 and older (M = 69.12, SD = 6.40) in a rural area of southern Taiwan completed all interview questions. The Chinese versions of the Personal Resource Questionnaire (PRQ) and the Positive Affect and Negative Affect Scale (PANAS) were used to measure social support and well-being. Content validity of the Chinese versions of the instruments was estimated as satisfactory. The alpha reliability coefficients were .86 for the entire PRQ-85 Part 2 and from .65 to .78 for the subscales. The alpha reliability coefficients were .85 and .65 for the PA subscale and NA subscale respectively. Pearson correlation coefficients were used to examine the relationship between social support and well-being. The findings demonstrated that not only the entire social support but also its five dimensions were positively correlated with positive affect (r ranged from .24 to .48, p < .01) and were inversely correlated with negative affect (r ranged from -.15 to -.21, p < .05 or .01). A substantive intervention design may help verify the effectiveness of social support on elderly women's positive affect and negative affect. Using social support as a strategy to promote these women's well-being, therefore, should be the subject of future study. PMID- 11392104 TI - Congenital rhabdomyosarcoma--a case report. AB - A term female newborn was noted to have a tumor mass in the oral cavity soon after birth. Oral computer tomography revealed a well-enhanced soft tissue mass about 4 x 4 x 3 cm in size in the left buccal area. Group III embryonal type congenital rhabdomyosarcoma was diagnosed after biopsy (gross removal was not feasible). Respiratory distress exacerbated due to rapid tumor growth compressing airway with the result that endotracheal tube had to be intubated. Chemotherapy was done and complicated by two episodes of neutropenic fever and sepsis. Radiotherapy was suggested but refused by the family. Tumor size was slightly reduced and endotracheal tube could be removed four months later. She was taken home under regular chemotherapy. Radiotherapy, was, however, clearly indicated. PMID- 11392105 TI - Brachial plexus injury following coronary artery bypass surgery: a case report. AB - Postoperative brachial plexus injury, often manifesting as a variety of upper extremity neuropathies, is a recognized and not uncommon complication following cardiac surgery that requires a median sternotomy. In general, the vast majority of its neurological symptoms are transient and need no treatment. Nevertheless, in very rare cases, the peripheral neuropathies will persist and cause disability. We treated a 67-year-old male patient complicated by permanent paresthesia and paralysis of the left upper extremity after an eventful coronary artery bypass surgery. The nerve conduction measurements and electromyography all revealed a C5 to T1 lesion. After carefully reviewing the surgical course and referring to the published literature, we tentatively concluded that compression or overstretching produced by wide and prolonged sternal separation of the brachial plexus was the most likely etiology. Asymmetrical traction of the sternal halves during internal mammary artery harvesting might also have contributed to this nerve injury. We surmised, therefore, that brachial plexus injury could be minimized by an exact median sternotomy, a lower position and the smallest possible opening for the sternal retractor, and the avoidance of constant and asymmetrical traction on the sternal halves. PMID- 11392106 TI - Struma ovarii mimicking a benign multicystic ovarian tumor: MR imaging in one case. AB - Struma ovarii is a rare form of mature teratoma of the ovary. A case of struma ovarii is presented with magnetic resonance (MR) imaging and pathologic findings. MR imaging showed a multiloculated cystic adnexal mass with well-defined margin and septum. Internal cystic spaces presented homogeneous signal intensity of fluid content in both T1-weighted and T2-weighted images. Subtle contrast enhancement was depicted in internal septi. Imaging features were indistinguishable from benign multicystic ovarian tumor, and different from the reported cases in the literature. PMID- 11392107 TI - Model for regulation of non-muscle myosins. AB - Diverse morphological changes of non-muscle eukaryotic cell are usually accompanied by dynamic remodeling in its cytoskeleton. The conformation/functional state of myosins is presumed to be markedly altered during cytoskeletal reorganization. Although myosins play critical roles in various forms of cellular movement and shape changes, the molecular basis for regulation of their activity is complicated and far from understood. A numerous experimental data show the phosphorylatable light chain (20 kDa), known as a regulatory light chain (RLC), as one of the key modulators of regulatory signals in the structure of myosin-II molecule. However, the clue that coordinates its phosphorylation state remains to be enigmatic. In the present work, we propose the most rationale mechanism which might regulate the state of RLC phosphorylation, and therefore, might control organization and activity of non muscle myosins. PMID- 11392108 TI - [Use of the acoustic startle response in the mouse to evaluate the pharmacodynamic action of ethanol]. AB - A method of the pharmacological screening of psychotropic drugs that is based on the estimation of the peak amplitude and latency of the acoustic startle response (ASR) is suggested. A linear relationship between the changes of the ASR parameters induced by acute i.p. 14C-ethanol administration and its contents in brain is demonstrated. It is proposed that changes of the ASR parameters are strictly dependent upon the concentration of a modulatory drug in the biophase of action (brain, serum) and therefore can be utilised for the effector analysis of pharmacokinetics and preliminary screening of psychotropic drugs. PMID- 11392109 TI - [Cardioprotective effects of a new fluorine-containing synthetic activator of ATP dependent potassium channels]. AB - Effects of PF-10 the new fluorine-containing activator of ATP-dependent K channels were investigated on the guinea-pig isolated heart. PF-10 enhanced myocardial contractility and decreased coronary vessels resistance in a dose dependent manner. Low doses (10 mM/L, 1 mM/L) of PF-10 attenuates of the spontaneous alterations of heart rate, while high doses (0.1 mM/L) leds to these alterations. Previous activation of ATP-dependent K-channels by PF-10 has a cardioprotective effect during ischemia-reperfusion which a similar to the well known phenomenon of ischemic preconditioning. PMID- 11392110 TI - [Effects of sex hormones on HERG potassium channels expressed in Xenopus oocytes]. AB - The repolarisation phase of cardiac action potential is characterized by sexual dimorphism suggesting the role of sex steroid hormones in the regulation of K+ channels. Here we report on the effects of testosterone and 17 beta-estradiol on HERG-encoded K+ channels, expressed in Xenopus oocytes. At 1M concentration testosterone decreased the amplitude of HERG-directed IKr (rapid component of cardiac delayed rectifier K+ current) by 30% within 30 min of exposure, while 17 estradiol had no effect. Testosterone did not alter the HERG channels kinetics, voltage-dependence, steady-state activation and inactivation properties suggesting that its action most probably involves the decrease of channels open probability and/or the level of their expression. The signaling pathways mediating testosterone-induced down-regulation of HERG channels in Xenopus oocytes remains to be elucidated. The effect of testosterone on HERG channels in Xenopus oocytes is opposite to what one might expect from shorter cardiac action potential duration and electrocardiographic QT-interval in males compared to females. PMID- 11392111 TI - [From Bogomoletz anti-reticular cytotoxic serum to monoclonal antibodies]. AB - The paper devoted to the 120th anniversary of A.A. Bogomoletz birth deals with the developmental stages of the cytotoxin (xenogenic antibody) doctrine from creating the anti-reticular cytotoxic serum (ACS) of Bogomoletz to the use of monoclonal antibodies both in biological investigations and in medicine. The use of ACS for treating various diseases and the mechanisms of its effect; obtaining other anti-organ and anti-tissue cytotoxic sera and establishing their experimental effects; the use of a set of cytotoxic sera, taken in both inhibitory and stimulating doses, in medicine and agriculture, as well as the drawbacks of cytotoxic sere (polyclonal xenogenic antibodies) are described. The experience of using monoclonal antibodies to identify the structures of protein antigenic determinants and to establish their role in different diseases are analysed. The use of monoclonal antibodies for treating various diseases is reviewed. The advances in the techniques of obtaining monoclonal antibodies are noted. PMID- 11392112 TI - [Ionic currents responsible for activation of isolated retinal ganglion cells]. AB - Ionic currents responsible for excitability of retinal ganglion cells (RGC) freshly isolated from eyes of adult rats were studied. Kutward potassium currents in RGC represented by populations of slow inactivating and non-inactivating currents. Inward sodium currents were completely blocked by extracellular application of 0.25 mM TTX. Calcium conductance in freshly isolated RGC has two components: low- and high-threshold. Extracellular application of nifedipine (10 mM) or Cd2+ (10 mM) blocked high threshold calcium currents by 28% and 60%, correspondingly, without any significant effect on low threshold calcium currents. PMID- 11392113 TI - [The effect of iron and zinc on lead absorption in the tunica mucosa of various parts of rat small intestine ]. AB - Kuring experiments on turned out sacs models of duodenum, jejunum and ileum of rats the differences in levels of exogenous Pb2+ absorption were revealed. The most values were in tunica mucosa (TM) of duodenum and jejunum. Pb2+ absorption was influenced by previous exposure to trace elements (TE) of Fe2+ (0.79 mmole/100 g) and Zn2+ (0.36 mmole/100 g) having been inducted to stomach. TE due to their physical and chemical properties reduced Pb2+ absorption in different parts of intestine TM on the average by 14-49%. It was determined that TE prevented Pb2+ absorption in TM of intestine more effectively during combined introduction compared to isolated one (accordingly 42% in duodenum, 43% in ileum and 29% in jejunum). Reducing Pb+ absorption is explained by probable substrates competition for active centers of their binding and transfer to TM to intestine. PMID- 11392114 TI - [The effect of L-arginine on the development of experimental cardiac hyperfunction and hypertrophy]. AB - It was shown, in the investigation on laboratory rats with experimental aortic coarctation, the development of myocardial hyperfunction and hypertrophy before and under the influence of the N kappa--predecessor--L-arginine. It was found that on the 14-20 day after coarctation weight of the heart, especially of the left ventricle, and contractive activity of the papillar muscle realistically increase. Under durable infusion of L-arginine to the rats (100 mg/kg) with aortic coarctation, weight of heart and myocardial contractive activity increase more significantly than without it. N kappa--predecessor secures intracellular structures of the myocardium. PMID- 11392115 TI - [Effect of nitric oxide synthase inhibitor L-NNA on the activities of antioxidant enzymes and lipid peroxidation in blood and tissues of rats with different resistance to hypoxia]. AB - The main components of antioxidant enzyme system (AOS) are superoxide dismutase (SOD) and glutathione reductase (GR) catalyses the conversion of the superoxide anion. The important role in AOS belongs to catalase and glutathione peroxidase which perform H2O2 to nontoxic products. Simultaneous determination of AOS activity and malonic dialdehide (MDA) concentration (the index of lipid peroxidation in tissues and blood) characterize cells complex resistance to damage factor. The effect of L-arginine, as a precursor of nitric oxide synthesis and blocator NO-synthase (Nw-nitro-L-arginine) on AOS of rats with different resistance to hypoxia under stress condition is unknown and were subject of our investigation. Experiments were done on liver and blood tissues of white laboratory rats. The experimental animals were divided on two groups depending on hypoxia factor: high resistance (HR) and low resistance (LR). The type of resistance was determined by the time of ability to respire in barocamera with oxygen deficient equal to 12.000 meters over sea level. The animals adaptation to laboratory conditions continue during 14 days after in barocamera presence. All animals were divided dependent on experiment conditions on fourth groups. The first group: intact (HR and LR) animals parentherally injected by 1 ml of 0.9% NaCl solution. The second group was subject of stress condition. The third group: HR and LR animals injected parentherally by 1 ml L-arginine (Sigma, USA) dose (600 mg/kg body weight). The fourth one: rats injected by 1 ml Nw-nitro-L arginine (L-NNA, Sigma, USA)--the blocator of NO-synthase. The animals were decapitated 30 min after injection and stress condition under ethereal anesthesia. Activity of antioxidant system enzymes superoxide dismutase (SOD), catalase (CAT); glutathione reductase (GR), glutathione peroxidase (GP) were measured spectrophotometrically. Also was investigated the concentration of serum antioxidant ceruloplasmin (CP). Level of lipid peroxidation was estimate by examination of concentration of lipids of hydroperoxides (LHP) and malonic dialdehyde (MDA). Our data confirm suggesting that nitric oxide (NO) is a major regulator in the AOS enzymes activity and limit damage influence of AOF. Action precursor NO L-arginine might be capable of protective role in various disorders which are connected with hypoxia factor. Following thing can be interred the investigation of influence of nitric oxide adaptive answers in stress condition modelling of pathological processes in rats with different resistance to hypoxia and reflect the biological qualities data stady on AOZ and LP. PMID- 11392116 TI - [The effect of ionizing radiation on the dynamics of lipid peroxidation final products in animal blood]. AB - The dynamics of dose-dependence of malonic dialdehyde (MD) in the blood serum of Vistar line male rats after a total single irradiation with 60Co gamma-quantums in 0.2; 0.5; 1.0; 3.0 and 5.0 Gy doses (during 120 days); 7.0 Gy (during 20 days); 90 Gy (during 15 days) were studied. MD contents values both in the norm and in 0.5, 1, 2, 4, 6, 8, 10, 15, 20, 30, 45, 65, 90 and 120 days after irradiation are given. The experimental data show that the dose dependent LPO final products have an wavelike character. PMID- 11392117 TI - [The effect of physiological and extreme irritants on zinc metabolism in pancreatic islets and hippocampus cells]. AB - Zinc content in pancreatic islet cells and hippocampus was studied with the aid of highly sensitive selective cytochemical reaction of 8-(p-toluensulfonylamino) quinoline. It was shown that the factors stimulating B-insulocyte secretory activity induced the decrease of this metal concentrations in the cell. An increase of these cells zinc content observed in the cases of their secretory activity inhibition. The extreme irritants induced two phase changes of zinc concentrations in pancreatic islets and hippocampus. PMID- 11392118 TI - [Spectral analysis of heart rate responses to light and color stimulation of the cerebral hemispheres]. AB - To study changes autonomic nervous function at differential light stimulations of the brain hemispheres in 126 healthy persons stimulated of the right and left hemifields in retinas eyes by light of different colour. Before and after influence the spectral analysis of a heart rhythm was made. The decrease of capacity of a spectral range 0.08-0.12 Hz is revealed at red-green stimulations, that testifies to decrease baroreflex tone and range 0.15-0.5 Hz at red-green and red-violet stimulations, that testifies about vagolitical influence. The conclusion about domination of the right hemisphere in autonomic nervous function is made. PMID- 11392119 TI - [Reciprocal interaction in human forearm muscles during passive and active rest]. AB - The reciprocal interaction in the forearm flexor in the human subjects were determined in the time of passive and active rest, using H-reflex testing. H reflex was evoked by stimulation of n. medianus after 1, 10 and 150 ms after conditioned stimulus. The maximum of reciprocal inhibition was found on the beginning of passive rest; however, recover motoneurones activity coincided in the suppression of this inhibition. The recovery of motoneurones' responses during active rest was realized through increase of three phases of reciprocal inhibition. The possible mechanisms of the studied phenomenon were discussed. PMID- 11392120 TI - [Comparative analysis of the effects of alcoholism and opium addiction on liver function]. AB - Groups of patients suffering alcoholism and narcomania were examined for the effect of intoxication on the blood serum enzymes of mainly liver origin: alcohol dehydrogenase (ADH), aldehyde dehydrogenase (ALDH), aspartate aminotransferase (AST), alanine aminotransferase (ALT), as well as on thymol test. It has been shown that in patients with the first stage of alcoholism one could observe only functional disturbances in the liver: the increase of ADH activity which evidences for the induction of its synthesis. In patients with the first stage of opium narcomania one can record total hyperenzymenia, decrease of de-Rimis coefficient at the expense of more considerable increase of ALT activity than that of AST, as well as the sharp increase of thymol test--these are the signs of destructive and metabolic disturbances in the liver. In patients with the second stage of alcoholism one can observe the decrease of ALDH activity under the increase of ADH, AST, ALT activity and high thymol test-these are the signs of toxical hepatitis. Destructive and metabolic changes increase in the liver in the patients with the second stage of narcomania. PMID- 11392121 TI - [Functional state of the body and mental capability in humans of various ages]. AB - The aim of investigation was to studied of functional state and mental capability of human in different age. The 150 women and 125 men in five age-groups: 13-18, 19-29, 30-39, 40-49, 50-60 [symbol: see text] were examined. The mental capability was modeled by special computer's test. The method of determination of functional age as a functional state were studied. The results was showed about decline of functional state with decrease of mental capability. The higher levels of mental capability are correspond to increasing of physical development rate in youth and decreasing of aging rate in adult. Was showed that compensation mechanisms of aging mental capability decrease are relation with activity of adaptational-regulatory physiological mechanisms vitaukt. PMID- 11392122 TI - [Ovarian function in androgenized rats in puberty]. AB - In experiment on rats injected with aromatase inhibitor or with dihydrotestosterone (DHT) it has been revealed that increase of androgens level in blood stimulates ovarial folliculogenesis on its early stages. DHT as compared with testosterone accelerates small follicles growth more effectively. Androgens, particularly DHT (7 mkg/100 g), suppress preovarial follicules maturing and turn them into atretic ones. Suppressing of ovarial functional activity delays or blockages the first ovulation during rats puberty. PMID- 11392123 TI - Do polychlorinated biphenyls contribute to reproduction effects in fish-eating birds? PMID- 11392124 TI - Monitoring of endocrine disruptors in surface waters by the yeast recombinant assay. AB - Endocrine disruptors exert physiological effects at very low concentrations. Surface waters present often a mixture of high concentrations of low-potency disruptors and low amounts of very powerful ones, making their chemical analysis complicated and expensive. We developed a recombinant yeast assay (RYA) for estrogenic compounds using 96-well microtiter plates. This assay is based on three yeast strains, transformed with self-propagating plasmids. One strain contains an expression plasmid for the human estrogen hormone receptor and an appropriate reporter; it detects estrogenic and antiestrogenic activities. The two other yeast strains, one expressing the human progesterone receptor and a second based on the yeast activator Gal4p, served to analyze the nature of antiestrogenic activities. We applied this technique to water samples from two tributaries on the Llobregat river (NE Spain) as well as from four sewage treatment plants discharging on them. Our results indicate that the efficiency of sewage treatment plants for eliminating estrogenic compounds varied notably, being in at least one case completely inefficient. We also observed a prevalence of an inhibitory activity all through the two rivers; this inhibition was hormone specific. These results were consistent to previously obtained chemical analyses of the same samples, although chemical and in vivo analyses showed rather different levels of sensitivity for some compounds. Our findings demonstrate the utility of the yeast recombinant assay for analyzing complex natural samples; at the same time, they stress the necessity of a panel of different yeast systems to adequately describe endocrine-disruptor activities. PMID- 11392125 TI - Effects of natural organic matter source on reducing metal toxicity to rainbow trout (Oncorhynchus mykiss) and on metal binding to their gills. AB - Juvenile rainbow trout (Oncorhynchus mykiss, 3 g) were exposed for 74 h in ion poor (soft) water to a mixed-metal solution in the presence of 4, 6, and 10 mg C/L natural organic matter (NOM). The metals were 0.2 microM Pb, 0.1 microM Hg, 0.1 microM Cd, 1.3 microM Cu, 0.05 microM Ag, and 3.5 microM Co, and the natural organic matter was isolated by reverse osmosis from three sources in southern Ontario, Canada. The six-metal solution alone was extremely toxic to the fish. Increasing concentrations of each NOM increased trout survival, but the NOM having the most allochthonous properties (from Luther Marsh) increased fish survival most, while the NOM having the most autochthonous properties (from Sanctuary Pond, Point Pelee) increased fish survival least. This pattern was reflected in the degree of reduction of Pb and Cu accumulation by the gills. Relatively simple chemical characterization of NOM, such as protein-to carbohydrate ratios, or optical characterization, such as absorbance-to fluorescence ratios (e.g., representing aromaticity), may adequately reflect these biologically relevant differences in organic matter quality. PMID- 11392126 TI - Indirect effects of zinc on soil microbes via a keystone enchytraeid species. AB - Effects of Zn pollution on a keystone species of forest soils, the enchytraeid Cognettia sphagnetorum, and its consequent indirect effects on microbial biomass and activity were studied in a microcosm experiment using experimentally contaminated humic soil. Microbial growth and decomposition were enhanced in the presence of C. sphagnetorum. At high Zn concentrations (< 2,393 mg/kg dry soil), populations of C. sphagnetorum went extinct, resulting in negative indirect effects on microbial activity as measured by soil respiration. Results indicate that pollution may affect species interactions in a soil food web and indirectly affect ecosystem processes such as decomposition rate. Effects of pollution on keystone organisms may radically alter soil ecosystem functioning and should be taken into account during risk-assessment procedures. PMID- 11392127 TI - Bioaccumulation of polycyclic aromatic compounds: 1. Bioconcentration in two marine species and in semipermeable membrane devices during chronic exposure to dispersed crude oil. AB - Assessing the fate in marine biota of hydrocarbons derived from oil particles that are discharged during exploration and production is of relevant environmental concern. However, a rather complex experimental setup is required to carry out such investigations. In this study, a sophisticated tool, the continuous-flow system (CFS), was used to mimic dispersed oil exposure to marine biota. Polycyclic aromatic hydrocarbon (PAH) uptake was studied in two species, the blue mussel Mytilus edulis and juvenile of the turbot Scophthalmus maximus, and in semipermeable membrane devices (SPMD) exposed to crude oil dispersed in a flow-through system. After an exposure period of 8 to 21 d, elimination in organisms and devices was analyzed for 9 to 10 d following transfer to PAH-free seawater. Principal component analysis (PCA) revealed different PAH patterns. In mussel and SPMD, the PAH profiles were very close to that analyzed in seawater. Slight differences were, however, indicated for large molecules with log Kow above six. Nonachievement of steady-state concentration and bioavailability of PAH in oil droplets may account for these differences. The PAH composition in fish revealed only congeners with two to three aromatic rings. A combination of bioavailability and efficient metabolism of the larger PAH molecules may explain this pattern. The CFS made possible a better understanding of some critical factors governing bioconcentration in marine biota from dispersed oil. Yet the results illustrate that uptake of PAH from exposure to oil particles is complex and that different species may bioconcentrate different molecules depending on factors like life style and metabolic capability to degrade the potential harmful substances. Hence, risk assessment of the actual impact of discharges to marine biota should consider these essential biological and ecological factors. PMID- 11392128 TI - Bioaccumulation of polycyclic aromatic compounds: 2. Modeling bioaccumulation in marine organisms chronically exposed to dispersed oil. AB - Within the frame of a large environmental study, we report on a research program that investigated the potential for bioaccumulation and subsequent effect responses in several marine organisms exposed to chronic levels of dispersed crude oil. Body burden can be estimated from kinetic parameters (rate constants for uptake and elimination), and appropriate body burden-effect relationships may improve assessments of environmental risks or the potential for such outcomes following chronic discharges at sea. We conducted a series of experiments in a flow-through system to describe the bioaccumulation kinetics of polycyclic aromatic hydrocarbons (PAH) at low concentrations of dispersed crude oils. Mussels (Mytilus edulis) and juvenile turbot (Scophthalmus maximus) were exposed for periods ranging from 8 to 21 d. Postexposure, the organisms were kept for a period of 9 to 10 d in running seawater to study elimination processes. Rate constants of uptake (k1) and elimination (k2) of the PAHs during and following exposure were calculated using a first-order kinetic model that assumed a decrease of the substances in the environment over time. The estimated bioconcentration factor was calculated from the ratio of k1/k2. The kinetic parameters of two-, three-, and four-ring PAHs in mussel and fish are compared with estimates based on hydrophobicity alone, expressed by the octanol-water partition coefficient, Kow (partitioning theory). A combination of reduced bioavailability of PAHs from oil droplets and degradation processes of PAHs in body tissues seems to explain discrepancies between kinetic rates based on Kow and actual kinetic rates measured in fish. Mussels showed a pattern more in compliance with the partitioning theory. PMID- 11392129 TI - Tumor prevalence and biomarkers of exposure in brown bullheads (Ameiurus nebulosus) from the tidal Potomac River, USA, watershed. AB - Associations between contaminant exposure and liver and skin tumor prevalence were evaluated in brown bullheads (Ameiurus nebulosus) from the tidal Potomac River, USA, watershed. Thirty bullheads (> or = age 3) were collected from Quantico embayment, near a Superfund site that released organochlorine contaminants; Neabsco Creek, a tributary with petroleum inputs from runoff and marinas; and Anacostia River (spring and fall), an urban tributary designated as a Chesapeake Bay region of concern, that was contaminated with polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and organochlorine pesticides. Fish were collected from the Tuckahoe River, as a reference. Cytochrome P450 activity, bile PAH metabolites, and muscle organochlorine pesticide and PCB concentrations were measured in randomly selected individuals and sediment contaminants were analyzed. We found statistically significant differences in liver tumor prevalences: Anacostia (spring), 50%; Anacostia (fall), 60%; Neabsco, 17%; Quantico, 7%; and Tuckahoe, 10%. Skin tumor prevalences were significantly different: Anacostia (spring), 37%; Anacostia (fall), 10%; Neabsco, 3%; Quantico, 3%; and Tuckahoe, 0%. Tumor prevalence in Anacostia bullheads warrants concern and was similar to those at highly contaminated sites in the Great Lakes. Evidence was found of higher PAH exposure in Anacostia fish but a cause-effect linkage could not be established. Fish tumor surveys, with histopathologic examination of internal and external organs, are recommended for monitoring the status of regions of concern. PMID- 11392130 TI - Simultaneous experimental study of direct and direct plus trophic contamination of the crayfish Astacus astacus by inorganic mercury and methylmercury. AB - An experimental study was carried out to investigate direct and direct plus trophic contamination routes of the crayfish Astacus astacus by inorganic mercury (Hg(II)) or methylmercury (MeHg). Direct exposure was based on low contamination conditions, 300 and 30 ng/L in the dissolved phase, respectively, during 30 d at 20 degrees C. Trophic exposure was based on daily consumption of the Asiatic clam Corbicula fluminea, previously contaminated during 40 d with similar exposure conditions. The Hg concentrations in the bivalves were very similar: 1,451 +/- 287 ng/g for Hg(II) and 1,346 +/- 143 ng/g for MeHg. In the crustaceans, Hg bioaccumulation was analyzed at the whole-organism level and in eight organs (gills, stomach, intestine, hepatopancreas, tail muscle, green gland, carapace, and hemolymph), after 15 and 30 d of exposure. Analysis of the results showed marked differences between Hg(II) and MeHg accumulation in favor of MeHg: for the direct route, the ratio between metal concentrations was close to 8; for the trophic route, no significant increase in Hg accumulation was observed for Hg(II) even when the ratio between Hg concentration in the direct plus trophic contamination route and Hg concentration in the direct contamination route was 1.6 for MeHg, with an estimated trophic transfer rate close to 20%. Mercury organotropism was also specifically connected to the exposure conditions, especially at the biological barrier level according to the route of exposure: gills and carapace for the direct route and digestive tract including hepatopancreas for the trophic route. PMID- 11392131 TI - Effects of the synthetic estrogen 17 alpha-ethinylestradiol on the life-cycle of the fathead minnow (Pimephales promelas). AB - A fish full life-cycle (FFLC) study was conducted for 17 alpha-ethinylestradiol (EE2) using the fathead minnow, Pimephales promelas. Newly fertilized embryos (< 24 h old) were exposed to five concentrations of EE2 (0.2, 1.0, 4.0, 16, and 64 ng/L nominal) in continuous flow-through conditions for 305 d at 25 +/- 1 degrees C. Exposure concentrations were verified by 14C-EE2 radiochemistry, supported by radioimmunoassay, and mean measured values were > or = 70% of nominal. For the F0 adult phase until 301 d posthatch, the no-observed-effect concentrations (NOECs) for growth, survival, and reproduction (as egg production) were all > or = 1.0 ng/L. The NOEC values for F1 embryo hatching success and larval survival (at 28 d posthatch) were both > or = 1.0 ng/L. While statistically detectable changes in F1 growth were evident at 0.2 ng/L, these were not considered to be biologically significant when compared with historical control data. Male fish exposed to EE2 at 4.0 ng/L failed to develop normal secondary sexual characteristics; on the other hand, assumed females exposed to this level of EE2 were able to breed when paired with males that had not been exposed to EE2. Histology of F0 control, 0.2 , and 1-ng/L exposed fish at 56 d posthatch indicated an approximate female-to male (F:M) sex ratio of 50:50 (with no ovatestes observed in the control), while fish exposed to EE2 at 4.0 ng/L for 56 d posthatch had a F:M sex ratio of 84:5 (with ovatestes in 11% of fish). After 172 d posthatch, no testicular tissue was observed in any fish exposed to EE2 at 4.0 ng/L. At the same time point, plasma vitellogenin levels were significantly higher in fish exposed to EE2 at 16 ng/L. A lack of sexual differentiation occurred in males at concentrations > or = 4.0 ng/L. Taking into account these data, the overall no-observed-adverse-effect concentration was considered to be 1.0 ng/L. PMID- 11392132 TI - Effects of Anabaena spiroides (Cyanobacteria) aqueous extracts on the acetylcholinesterase activity of aquatic species. AB - The effects of aqueous extracts from a cyanobacteria species, Anabaena spiroides, on fish (Odontesthes argentinensis), crab (Callinectes sapidus), and purified eel acetylcholinesterase (AChE) activity were studied. In vitro concentrations of A. spiroides aqueous extract that inhibited 50% of enzyme activity (IC50) were 23.0, 17.2, and 45.0 mg/L of lyophilized cyanobacteria for eel, fish, and crab AChE, respectively. Eel AChE inhibition follows pseudo-first-order kinetics, the same expected for organophosphorus pesticides. Inhibition of purified eel AChE using mixtures of bioxidized malathion and aqueous extract of A. spiroides showed a competitive feature (p < 0.05), suggesting that the toxin(s) could be structurally similar to an organophosphorus pesticide and that toxins present in the aqueous extract inhibit the active site of the enzyme. The inhibition recovery assays using 2-PAM (0.3 mM) showed that (1) bioxidized malathion inhibited 27.0 +/- 1.1% of crab and 36.5 +/- 0.1% of eel AChE activities; (2) with bioxidized malathion + 2-PAM the registered inhibition was 13.2 +/- 2.1% and 3.7 +/- 0.5% in crab and eel AChE, respectively; (3) the aqueous extract from A. spiroides inhibited 17.4 +/- 2.2% and 59.9 +/- 0.5% of crab and eel AChE activity, respectively; and (4) aqueous extract + 2-PAM inhibited 22.3 +/- 2.6 and 61.5 +/- 0.2% of crab and eel AChEs. The absence of enzyme activity recovery after 2-PAM exposure could imply that the enzyme aging process was extremely quick. PMID- 11392133 TI - Stable strontium accumulation by earthworms: a paradigm for radiostrontium interactions with its cationic analogue, calcium. AB - The accumulation of stable strontium and its chemical analogue calcium by four species of earthworm, representing three distinct ecophysiological groups inhabiting celestite (SrSO4)-rich natural soils, was investigated. An increase in soil strontium concentration over a four-orders-of-magnitude range was accompanied by an increase in earthworm tissue strontium concentration. In contrast, no relationship between soil and tissue calcium concentrations was apparent, indicating that this essential cation is strongly regulated within earthworm tissues but that nonessential strontium burdens are not physiologically regulated. Concentration factors for strontium ranged from 0.1 to 1.42, with an inverse relationship between this parameter and soil strontium concentration. The use of concentration factors and discrimination coefficients lead to the conclusion that earthworms discriminate against strontium accumulation at increased soil calcium concentrations, such that strontium would be predicted to be more efficiently accumulated by worms inhabiting acidic, calcium-poor soils. Although few consistent trends in strontium and calcium concentration were apparent between earthworm species, the species with the highest calcium turnover (i.e., the litter-inhabiting Lumbricus rubellus) typically accumulated higher concentrations of strontium than representatives of contrasting ecophysiological classes. These findings are considered in the context of the distribution, retention, and cycling of radiostrontium in soils. PMID- 11392134 TI - Ethoxyresorufin-O-deethylase induction in trout exposed to mixtures of polycyclic aromatic hydrocarbons. AB - This study investigated whether ethoxyresorufin-O-deethylase (EROD) activity in rainbow trout exposed to mixtures of polycyclic aromatic hydrocarbons (PAHs) could be predicted from induction equivalency factors (IEF). The test PAHs were classified into strong and weak inducers on the basis of similar exposure response curves. Induction equivalency factors of strong inducers, based on benzo[k]fluoranthene (BkF) as the reference compound, ranged from 0.03 to 0.16. Trout exposed to mixtures of strong inducers (2, 4, and 6 equipotent parts) at 0.32-, 1.0-, or 3.2-nM BkF-equivalents showed exposure-dependent increases in EROD activity, consistent with an additive interaction. The extent of activity did not vary greatly among mixtures and single PAHs at a given induction equivalent quantity (IEQ). Induction equivalency factors could not be calculated for weak inducers because the range of induction was too low. Hence, each weak inducer was added to mixtures at concentrations that induced EROD activity fivefold. These mixtures appeared additive because binary and quaternary mixtures caused about 10- and 20-fold induction, respectively. Strong inducers mixed the same way also showed additivity. In contrast, EROD induction by mixtures containing both strong and weak PAHs was 800 to 900% greater than expected, suggesting synergistic interactions. Therefore, if mixtures are composed of PAHs that behave similarly, IEFs may be a suitable approach for assessing risk. However, for mixtures that contain PAHs of differing potency and efficacy, bioassays will be a more reliable measure of risk than IEFs. PMID- 11392135 TI - A large-scale categorization of sites in San Francisco Bay, USA, based on the sediment quality triad, toxicity identification evaluations, and gradient studies. AB - Sediment quality was assessed in San Francisco Bay, California, USA, using a two tiered approach in which 111 sites were initially screened for sediment toxicity. Sites exhibiting toxicity were then resampled and analyzed for chemical contamination, recurrent toxicity, and, in some cases, benthic community impacts. Resulting data were compared with newly derived threshold values for each of the metrics in a triad-based weight-of-evidence evaluation. Sediment toxicity test results were compared with tolerance limits derived from reference site data, benthic community data were compared with threshold values for a relative benthic index based on the presence and abundance of pollution-tolerant and -sensitive taxa, and concentrations of chemicals and chemical mixtures were compared with sediment quality guideline-based thresholds. A total of 57 sites exceeded threshold values for at least one metric, and each site was categorized based on triad inferences. Nine sites were found to exhibit recurrent sediment toxicity associated with elevated contaminant concentrations, conditions that met program criteria for regulatory attention. Benthic community impacts were also observed at three of these sites, providing triad evidence of pollution-induced degradation. Multi- and univariate correlations indicated that chemical mixtures, heavy metals, chlordanes, and other organic compounds were associated with measured biological impacts in the Bay. Toxicity identification evaluations indicated that metals were responsible for pore-water toxicity to sea urchin larvae at two sites. Gradient studies indicated that the toxicity tests and benthic community metrics employed in the study predictably tracked concentrations of chemical mixtures in Bay sediments. PMID- 11392136 TI - Evaluation and use of sediment toxicity reference sites for statistical comparisons in regional assessments. AB - Sediment reference sites were used to establish toxicity standards against which to compare results from sites investigated in San Francisco Bay (California, USA) monitoring programs. The reference sites were selected on the basis of low concentrations of anthropogenic chemicals, distance from active contaminant sources, location in representative hydrographic areas of the Bay, and physical features characteristic of depositional areas (e.g., fine grain size and medium total organic carbon [TOC]). Five field-replicated sites in San Francisco Bay were evaluated over three seasons. Samples from each site were tested with nine toxicity test protocols and were analyzed for sediment grain size and concentrations of trace metals, trace organics, ammonia, hydrogen sulfide, and TOC. The candidate sites were found to have relatively low concentrations of measured chemicals and generally exhibited low toxicity. Toxicity data from the reference sites were then used to calculate numerical tolerance limits to be used as threshold values to determine which test sites had significantly higher toxicity than reference sites. Tolerance limits are presented for four standard test protocols, including solid-phase sediment tests with the amphipods Ampelisca abdita and Eohaustorius estuarius and sea urchin Strongylocentrotus purpuratus embryo/larval development tests in pore water and at the sediment-water interface (SWI). Tolerance limits delineating the lowest 10th percentile (0.10 quantile) of the reference site data distribution were 71% of the control response for Ampelisca, 70% for Eohaustorius, 94% for sea urchin embryos in pore water, and 87% for sea urchins embryos exposed at the SWI. The tolerance limits are discussed in terms of the critical values governing their calculation and the management implications arising from their use in determining elevated toxicity relative to reference conditions. PMID- 11392137 TI - Description and evaluation of a short-term reproduction test with the fathead minnow (Pimephales promelas). AB - Due to the time and expense associated with full life-cycle testing, most current toxicity tests with fish do not explicitly consider reproductive output as an endpoint but, rather, focus on early life-stage survival and development. However, some classes of chemicals could adversely impact reproduction at concentrations below those that affect development. Further, estimates of the effects of toxic compounds on reproductive output can be critical to the ecological risk assessment process. In this manuscript, we describe a short-term reproduction test with the fathead minnow (Pimephales promelas) and evaluate the test using two model reproductive toxicants, methoxychlor (an estrogenic compound) and methyltestosterone (an androgenic chemical). The test is initiated with reproductively mature animals and is comprised of a pre-exposure phase of 14 to 21 d, followed by a chemical exposure of up to 21 d. During and at completion of the test, several endpoints related to reproductive fitness and endocrine function are assessed. Both chemicals evaluated in our study caused a significant decrease in fecundity of the fish at nominal concentrations of 5.0 micrograms/L (methoxychlor) and 0.2 mg/L (methyltestosterone). Methoxychlor decreased plasma concentrations of one or more steroids (testosterone, 11-ketotestosterone, beta estradiol) in both sexes and caused a significant induction of plasma vitellogenin in males, a response consistent with activation of the estrogen receptor by the pesticide (or its metabolites). Methyltestosterone decreased plasma concentrations of sex steroids and adversely affected gonadal status (as evaluated by relative weight and histopathology) in both sexes. The androgenic nature of methyltestosterone was clearly expressed as masculinization of exposed females via formation of nuptial tubercles, structures normally present only in reproductively active males. The chemical also caused a significant induction of plasma vitellogenin in both males and females; this unexpected estrogenic response was most likely due to aromatization of the androgen to a form capable of binding to the estrogen receptor. These studies demonstrate the utility of this short-term assay for identifying chemicals that exert reproductive toxicity through alterations in endocrine systems controlled by estrogens and androgens. PMID- 11392138 TI - Comparison of laboratory and in situ sediment bioassays using Corophium volutator. AB - Bioassays with the marine amphipod Corophium volutator were performed simultaneously in situ and in the laboratory using sediments sampled from the in situ locations. In most cases, the in situ response was significantly higher compared to the laboratory response. This difference was not caused by direct influence of the use of the field chamber on Corophium sp., nor was the difference caused by the overlying water used. Experiments showed homogenization can affect the toxicity of a sediment, but not in such a way that it can completely explain the difference between the response in situ and in the laboratory. Possible explanatory factors are harbor activity, storms, and temperature. To reduce the influence of some of these factors, the best period of the year to perform in situ bioassays with C. volutator is May, June, or September. PMID- 11392139 TI - Factors affecting reproduction and the importance of adult size on reproductive output of the midge Chironomus tentans. AB - We conducted two separate tests to evaluate the influence of several factors that could affect estimation and interpretation of effects on reproductive output of Chironomus tentans in sediment toxicity tests. Specifically, the influence of adult size, mating frequency in males (male), and age of both males and females (female) at first mating on mating success (number of successful matings), fecundity (number of eggs/female), percentage hatch, and number of offspring (number of hatched eggs) was assessed. In the first experiment, the influence of adult size on reproductive output was determined by growing midges fed a low (0.29 mg/individual/d) and a high (0.42 mg/individual/d) amount of food to produce small (S) and large (L) adults, respectively. The adults were then mated in one of four scenarios: S male x S female, S male x L female, L male x S female, and L male x L female. An increase in male weight at a constant female weight had no significant effect on fecundity, whereas an increase in female weight yielded 49.5 and 60.7% increases in fecundity when mated with low- and high-fed males, respectively. Similarly, mean number of offspring (fecundity x percent hatch) increased by 19.8 and 48.3% when male weight was increased and female weight was held constant and by 141.9 and 180.4% when female weight was increased and male weight was held constant. In the second experiment, conducted at a single feeding rate, fecundity increased significantly with an increase in female weight but not male weight. The number of offspring was not significantly influenced by adult weight. The age of males or females at first mating, the frequency with which males were mated (up to seven consecutive days), and the proportion of males successfully mating had no effect on fecundity or number of offspring; however, large increases in variability about mean estimates were observed in some endpoints for older males. These results suggest that adult female size is the most important factor affecting reproductive output in C. tentans, and that enhanced performance of the life-cycle test with respect to all reproductive endpoints might be achieved if males are not mated for more than five consecutive days. PMID- 11392140 TI - Gill lesions and death of bluegill in an acid mine drainage mixing zone. AB - The toxicity of an acid mine drainage (AMD) mixing zone was investigated by placing bluegill (Lepomis macrochirus) at the confluence of a stream contaminated by AMD and a stream having neutral pH. A mixing channel receiving water from both streams was assembled in the field, during July and October 1996, to determine the toxicity of freshly mixed and aged water (2.9-7.5 min). The AMD stream had elevated concentrations of Al and Fe, which precipitated upon mixing, and of Mn, which did not precipitate in the mixing zone. Fish exposed to freshly mixed water had higher mortality than fish exposed to water after aging. Precipitating Al, but not Fe, accumulated on the gills of bluegill, and accumulation was more rapid early during the mixing process than after aging. Fish exposed for 3.5 h to freshly mixed water had hypertrophy and hyperplasia of gill filament and lamellar epithelial cells. Similar lesions were observed after 6.0 h in fish exposed to water aged after mixing. Results demonstrated that Al was the predominant metal accumulating on the gills of fish in this AMD mixing zone, and that mixing zones can be more toxic than AMD streams in equilibrium. PMID- 11392141 TI - Effects of nutrient loading and planktivory on the accumulation of organochlorine pesticides in aquatic food chains. AB - The effects of nutrients and planktivory on the accumulation of hydrophobic organic contaminants (HOCs) in aquatic food chains were investigated in large lake enclosures. Food-chain compositions in the enclosures were manipulated by additions of planktivorous fish (+F), nutrients (+N), both nutrients and fish (+NF), or received no additions (-NF). The treatments resulted in higher plankton but lower zooplankton biomass in the +NF enclosures than in the other enclosures. Once enclosure communities were established, a suite of organochlorine pesticides (alpha-hexachlorocyclohexane, methoxychlor, heptachlor, cis- and trans-chlordane, cis- and trans-nonachlor, and mirex) was added to all enclosures in amounts sufficient to obtain initial concentrations in the epilimnion of approximately 15 ng/L. Dissipation of HOCs from the water and accumulation in phytoplankton, zooplankton, and fish were monitored for four months. The HOC concentrations in plankton did not differ significantly across treatments. However, on a total-mass basis, greater amounts of HOCs were sorbed to phytoplankton in the +NF enclosures (20%) than in the three other sets of enclosures. Concentrations in zooplankton of some HOCs differed significantly between treatments as a function of nutrient loading. Chlordane and nonachlor concentrations were greater in zooplankton from enclosures with no fish (+N, -NF) than in those from enclosures with fish (+F, +NF). The HOC residues in fish were highest in low-nutrient enclosures. The results demonstrate that fish predation and nutrient loading can modify the size related processes of HOC partitioning and affect its accumulation in the aquatic food chain. PMID- 11392142 TI - Evaluation of basal micronucleus frequency and hexavalent chromium effects in fish erythrocytes. AB - Hexavalent chromium (Cr [VI]) genotoxicity was studied using fish micronucleus analysis in peripheral blood erythrocytes from Pimephales promelas, the fathead minnow. Forty-five- to 60-d-old fish were used to assess the spontaneous level of genotoxic damage. The genotoxic effect of Cr (VI) obtained from potassium dichromate (K2Cr2O7) in tests performed for 7-, 14-, and 21-d exposure periods was estimated. Significant micronucleated erythrocyte (MNE) induction was detected in fish exposed for 7 d to 2.5 mg/L of Cr (VI), and induction decreased after 21 d of exposure. The results suggest a handling effect in fish manipulated compared to those not manipulated, thus demonstrating the importance of including parallel negative controls in experimental design. Basal levels of MNE are reported, providing laboratory values for future assay quality control. The importance of determining the period with the highest expression of the genotoxic effects in this assay system was also confirmed. PMID- 11392143 TI - Identification of androstenedione in a river containing paper mill effluent. AB - Effluent from a paper mill discharging into the Fenholloway River, Taylor County, Florida, USA, contains chemicals that masculinize females of the resident population of eastern mosquitofish (Gambusia holbrooki), as evidenced in females by elongated anal fins, which is normally a male-specific trait. To identify androgenic components in the effluent, water collected from the Fenholloway River and a control tributary was fractionated using solid-phase extraction and reverse phase high-performance-liquid chromatography. Two Fenholloway River fractions induced androgen receptor-dependent transcriptional activity in transient transfection cell culture assays. Of these, androstenedione was confirmed by liquid chromatography-mass spectrometry with multiple reaction monitoring. PMID- 11392144 TI - Effects of eight polycyclic aromatic compounds on the survival and reproduction of the springtail Folsomia fimetaria L. (Collembola, isotomidae). AB - The effects of eight polycyclic aromatic compounds on the survival and reproduction of the collembolan Folsomia fimetaria L. were investigated in a well characterized Danish agricultural soil. With the exception of acridine, polycyclic aromatic hydrocarbons (PAHs) and neutral N-, S-, and O-monosubstituted analogues showed similar toxicities to soil collembolans when the results were expressed in relation to total soil concentrations (mg/kg). The estimated concentrations resulting in a 10% reduction of reproductive output (EC10 values) were based on measured initial concentrations and were for acridine 290 mg/kg, carbazole 10 mg/kg, dibenzofuran 19 mg/kg, dibenzothiophene 7.8 mg/kg, fluoranthene 37 mg/kg, fluorene 7.7 mg/kg, phenantrene 23 mg/kg, and pyrene 10 mg/kg. When the EC10 values were converted to soil pore-water concentrations, they showed a highly significant correlation (r2 = 0.71, p < 0.01) to no-observed effect concentrations for the freshwater crustacean Daphnia magna, as estimated by a quantitative structure activity relation (QSAR) for baseline toxicity (nonpolar narcosis). Only carbazole and acridine were more than two times more toxic (4.9 and 3.1, respectively) than expected from the Daphnia QSAR data. The latter result indicates that the toxicity of the tested substances is close to that expected for compounds with nonpolar narcosis as the mode of action. However, the relatively large uncertainties in the extrapolation method prevent final conclusions from being drawn. PMID- 11392145 TI - Effect of beringite on cadmium and zinc uptake by plants and earthworms: more than a liming effect? AB - Metal-contaminated soils are potentially harmful to plants, animals, and humans. Harmful effects are often related to the free-metal concentration in the soil solution. Immobilization is a potentially useful method to improve the quality of metal-contaminated soils by transforming free-metal ions into species that are less mobile and less toxic. The effect of many immobilizing products can be attributed to sorption on the surface of the material. Alkaline materials also enhance adsorption to soil particles by decreasing proton competition. Immobilization should preferably be evaluated independently of soil pH to discriminate between these processes. In this study, the immobilizing effect of beringite, an alkaline alumino silicate, was compared with that of lime. Plants (Swiss chard [Beta vulgaris L. var. cicla]) were grown on a soil contaminated with cadmium and zinc and treated with graded amounts of beringite or lime. Metal availability, as determined by a 0.01 M CaCl2 extraction, and metal uptake by plants strongly decreased in all treated soils. Beringite did not reduce metal availability more than liming when the obtained pH levels were similar. The effect of beringite can, therefore, be explained as a liming effect, at least for the duration of our experiment (10 weeks). The effect of beringite and lime on metal accumulation by earthworms (Eisenia veneta and Lumbricus rubellus) was small or not significant, although the CaCl2-extractable metal concentration in treated soils decreased by more than 90%. We conclude that immobilizing agents based on a liming effect can decrease metal uptake by plants, but they will hardly affect metal uptake by earthworms. Hence, these materials can reduce negative ecological effects of metal contamination on plants and herbivores, but not on earthworm predators. PMID- 11392146 TI - Effects of endophyte status of tall fescue tissues on the earthworm (Eisenia fetida). AB - A cryptic fungal endophyte, Neotyphodium coenophialum, infects most tall fescue (Festuca arundinacea) pastures in the United States. Cattle, sheep, and horses that consume the endophyte-infected grass can suffer fescue toxicosis caused by toxic alkaloids in the infected plants. The effects of the endophyte on mammalian herbivores have been well documented, but less is known regarding the quality of the grass (infected vs noninfected) as a food material for soil invertebrates. We conducted 21-d tests with earthworms (Eisenia fetida) to determine the nutritional quality of endophyte-infected and noninfected tall fescue leaf and root tissues. Earthworm survival, growth, and reproduction were measured in each treatment combination of tissue type and infection status. Earthworm survival was 100% in all treatments. Tall fescue tissue type (leaf vs root) and infection status (present or absent) both significantly affected E. fetida growth and reproduction. The earthworms grew and had moderate levels of reproduction in replicates containing tall fescue leaf tissue as the sole food source regardless of the endophyte-infection status, but earthworms lost weight and had less reproduction in replicates where tall fescue root tissue was the sole food source. An unexpected effect of infection status on earthworm growth in the tall fescue leaf-tissue treatments was also evident: mean growth of E. fetida with access to endophyte-infected leaf tissue as the sole food source was 3.6-fold greater than mean growth of E. fetida with access to noninfected leaf tissue as the sole food source. Knowledge regarding the relative effects of endophyte status of tall fescue on soil organisms may allow the development of more effective environmental management strategies at sites where tall fescue is being considered for use in phytoremediation or for vegetative cover. Investigators working with tall fescue should be alert to the possibility of endophyte-mediated effects of tall fescue on other organisms and, at a minimum, should provide information regarding the grass's infection status when reporting the results of studies that involve use of tall fescue. PMID- 11392147 TI - Synergistic effects of a photooxidized polycyclic aromatic hydrocarbon and copper on photosynthesis and plant growth: evidence that in vivo formation of reactive oxygen species is a mechanism of copper toxicity. AB - Heavy metals and polycyclic aromatic hydrocarbons (PAHs) are often cocontaminants in industrialized environments, yet little is known about either the extent or mechanisms of their cotoxicity. To address this shortfall, the combined effects of an oxygenated PAH, 1,2-dihydroxyanthraquinone (1,2-dhATQ), and a heavy metal, Cu2+, on photosynthesis and growth of the duckweed (Lemna gibba) were evaluated. Using assays of chlorophyll a fluorescence and photosystem I activity, 1,2-dhATQ inhibited electron transport at the cytochrome b6/f complex. Conversely, Cu2+ alone (at low concentrations) had little effect on photosynthesis. When Cu2+ was combined with 1,2-dhATQ, an increase in transient and steady-state chlorophyll a fluorescence quenching occurred relative to 1,2-dhATQ alone. Treatment of isolated thylakoid membranes with 1,2-dhATQ inhibited whole-chain linear electron transport, measured as O2 consumption using methyl viologen as the electron acceptor. However, Cu2+ plus 1,2-dhATQ resulted in active O2 consumption with or without methyl viologen as an electron acceptor. From these data, we conclude that 1,2-dhATQ renders the plastoquinone pool to a highly reduced state by inhibiting at cytochrome b6/f. Then, Cu2+ is able to mediate the transfer of electrons from reduced plastoquinone to O2, forming reactive oxygen species. At the whole-organism level, when Cu2+ and 1,2-dhATQ were mixed at concentrations that resulted in the above-mentioned impacts on photosynthesis, synergistic inhibition of plant growth was observed. This suggests a catalytic mechanism of toxicity for redox active metals, a process that could be instrumental in explaining their impacts at low concentrations. PMID- 11392148 TI - Pyrene bioaccumulation, effects of pyrene exposure on particle-size selection, and fecal pyrene content in the oligochaete Limnodrilus hoffmeisteri (Tubificidae, Oligochaeta). AB - The oligochaete Limnodrilus hoffmeisteri Cleparede was exposed to sediment amended pyrene (0-1,196 nmol/g dry wt) for 2, 5, and 10 d to investigate sediment particle-size preference, tissue pyrene bioaccumulation (using biota-sediment accumulation factor, BSAF), fecal pyrene concentrations (using fecal-sediment accumulation factor, FSAF), and pyrene biotransformation. In non-pyrene-amended sediment, L. hoffmeisteri preferentially ingested finer sediment particles (< 3.5 microns). However, pyrene concentrations > 199 nmol/g dry weight resulted in a decreased preference for finer particles, presumably reducing pyrene exposure. Particle selectivity also changed with time such that after 10 d, worms showed an increased preference for finer particles. At high pyrene exposure, tissue pyrene concentrations rapidly attained equilibrium (maximum body residue 4.4 mumol/g wet wt, close to values cited for hydrocarbon critical body residues). Levels of biotransformation were low (mean 14%) and independent of time or pyrene concentration. Biota-sediment accumulation factor showed a negative relationship with sediment pyrene concentration; this trend may be due to reduced availability at high sediment pyrene concentrations or due to reductions in both ingestion rate and selectivity for fine, pyrene-rich particles. Fecal pyrene concentrations were depleted compared to sediment levels, suggesting removal of pyrene during gut passage by absorption. Fecal-sediment accumulation factor showed a negative relationship with increasing sediment pyrene levels. PMID- 11392149 TI - Comparison of metal uptake rate and absorption efficiency in marine bivalves. AB - Recent studies have quantified extensively metal assimilation efficiency from ingested food sources in aquatic invertebrates. Metal absorption efficiency (alpha) from the dissolved phase is analogous to metal assimilation efficiency, but it remains poorly defined and quantified. In this study, the alpha of four trace metals [Cd, Cr(VI), Se(IV), and Zn] was determined in three species of marine bivalves (green mussel [Perna viridis], black mussel [Septifer virgatus], and clam [Ruditapes philippinarum]). Individual bivalves were first measured for their clearance rates, followed by measurements of the metal influx rate, after which the metal alpha and the uptake rate constant (Ku) were then computed. Among the four metals considered, the highest Ku and alpha were found for Zn, followed by Cd > Cr(VI) > Se(IV). The Ku values were comparable between the two mussels but were 1.8- to 3.3-fold lower in the clams. Interspecific difference in metal Ku was strongly related to, but intraspecific difference in Ku was not affected by, the bivalve's clearance rate. Interspecific difference in metal alpha was smaller than the metal Ku and was independent of the clearance rate, whereas the intraspecific difference in metal alpha correlated with the individual variations of the clearance rate. Within each bivalve species, a significant negative correlation was found between the metal alpha and the clearance rate, implying that an individual pumping a greater amount of water was coupled to a lower alpha. Significant correlation between the alpha of four metals was also documented in all three species of bivalves. Thus, metal bioavailability from the aqueous phase was directly related to the physiological conditions of the animals. Both the aqueous chemistry and the physiology of the animals can be important in affecting metal bioavailability from the dissolved phase. PMID- 11392151 TI - Inflammation, malnutrition, cardiovascular disease and mortality in end-stage renal disease. PMID- 11392150 TI - Toxicant- and response-specific comparisons of statistical methods for estimating effective concentrations. AB - Standard U.S. Environmental Protection Agency (U.S. EPA) statistical analyses of whole effluent toxicity tests involve the estimation of the concentration associated with a specified level of inhibition relative to control responses. Current U.S. EPA estimation methods (linear interpolation or probit-based methods) are compared to a recently developed parametric regression-based estimator, the relative inhibition concentration estimator RIp. The RIp estimation technique, with level of inhibition p = 25%, is applied to a series of chronic toxicity test data from a U.S. EPA Region 9 database of reference toxicity tests. Tests on marine species are conducted with one reference toxicant, while the freshwater tests are conducted with several reference toxicants. While the U.S. EPA estimators and the RIp estimator are highly correlated for red abalone larval shell development, the degree of correlation for fathead minnow responses varies with reference toxicant tested. The strength of the relationship between the RIp and the standard U.S. EPA estimators varies as a function of the reference toxicant. Correlations range between 0.67 and 0.99. For all biological responses included in this evaluation (fathead minnow growth or survival and red abalone larval development), experiments occurred where the RIp is estimable, while the standard U.S. EPA estimators are not. Nonestimability of the standard EPA methods appears to be related, in part, to the failure of models to account for enhanced responses, such as a hormesis effect prior to toxicity being manifest. The ability to account for such enhanced responses is a strength of the RIp method. Finally, a variance component analysis suggests that lab-to-lab variability is relatively low for the red abalone but relatively high for the fathead minnow. PMID- 11392152 TI - Prevention and treatment of the malnutrition, inflammation and atherosclerosis (MIA) syndrome in uremic patients. PMID- 11392153 TI - [Advances in treatment of hypertension]. PMID- 11392154 TI - [The role of hypertension in progression of chronic renal failure--therapeutic prospects]. PMID- 11392155 TI - [Pathogenesis, diagnosis and conservative treatment of ischemic heart disease in patients with chronic renal failure]. PMID- 11392156 TI - [Myocardial revascularization in patients with chronic renal failure: possibilities for interventional cardiology and cardiosurgery]. PMID- 11392157 TI - [Usefulness of extracorporeal blood purification methods in prevention and treatment of cardiovascular diseases]. PMID- 11392158 TI - [Hyperhomocysteinemia as a risk factor for cardiovascular diseases in patients with chronic renal failure]. PMID- 11392159 TI - [Ventricular arrhythmia in patients with chronic renal failure treated with hemodialysis]. AB - From the group of 66 patients (pts) treated with in-hospital haemodialysis (HD), 30 pts were selected for 48 hrs monitoring of heart rhythm to register arrhythmias using Holter method. Cardiovascular complications were observed in 24 pts (80%) of the studied group; ischemic heart disease in 10 pts (33%), chronic cardiac failure in 8 pts (27%), left ventricular hypertrophy in 16 pts (53%) and hypertension in 24 pts. During 48 hrs of heart rhythm monitoring ventricular heart arrhythmias (VHA) were registered in 23 pts. 8 pts of this group had more then 100 additional ventricular beats for 24 hrs. VHA were registered before HD in 14 pts, during HD in 15 pts and after in 15 pts. The frequency of VHA pt/one hour of monitoring increased during and immediately after HD. There were no statistically significant differences between 23 pts with VHA and 7 pts without VHA with respect to the following parameters measured before HD: blood pressure, urea, calcium, kalium and magnesium blood concentrations. We found statistically significant difference between both groups of pts for creatinine values (p < 0.02); respectively 899.7 mmol/l SD 152 mmol/l versus 767 mmol/l SD 95.3 mmol/l and for interdialytic body weight increase (p < 0.012); respectively 2.65 kg SD 0.8 kg versus 2.04 kg SD 0.46 kg. Our initial results indicate that VHA appears in the majority of hemodialysed pts and that HD intensifies arrhythmogenic influence of irreversible renal failure on heart. It is also possible that non adequate HD might be responsible for induction of ventricular heart arrhythmias during and after dialysis. PMID- 11392160 TI - [Efficacy of spiral computed tomography in evaluation of renal arteries in patients with renovascular hypertension]. AB - The aim of this study is an estimation of efficacy of spiral computed tomography (SCT) in evaluation renal arteries in patients with renovascular hypertension. SCT was performed in 54 patients (32 females and 22 males) by means of Picker PQ 2000. The thickness 4 mm, index 4 mm, pitch 1.0. Contrast agents-Uropolinum 75% or Ultravist 350 were applied automatically 3-4 mm/sec. with delay 20-22 sec. Secondary reconstruction with 2 mm leyers was performed by Maximum Intensity Projection (MIP). In all patients the abdominal arteriography was performed as a reference method. The coincidence of both methods in evaluation of renal arteries occlusion or stenosis was estimated by Cohen kappa coefficient. In addition, sensitivity and specificity of SCT were estimated. RESULTS: Consistence of SCT and arteriography for right, left and additional renal artery were as follows: 0.914, 0.92, 0.85. Sensitivity and specificity SCT for rigt, left and additional renal artery were as follows: 95.5% and 95.8%, 96.5% and 95.8%, 50% and 100%. The highest efficacy of SCT was noted in ostium, the most difficult place in evaluation of renal artery during arteriography examination. PMID- 11392161 TI - [The role of angiotensin I converting enzyme (ACE) Pst 1 polymorphism in the development of chronic renal failure]. AB - Chronic renal failure (CRF) is a complex phenotype, which results from the underlying kidney disease and superimposing environmental and genetic factors. The aim of the study was to evaluate the role of angiotensin converting enzyme gene Pst 1 polymorphism in the development and/or progression of CRF. 247 family trios (patients with CRF and both parents): 120 with primary chronic glomerulonephritis, 80 with interstitial nephritis and 47 with diabetic nephropathy were examined, and transmission/disequilibrium test (TDT) was used to evaluate allele transmission from heterozygous parents to affected off-spring. No significant deviation from random transmission of the examined ACE gene Pst 1 alleles was observed, as well as no impact of this marker on the rate of progression of chronic renal failure. PMID- 11392162 TI - [Does any relationship exist between metabolic disturbances and some markers of renal damage in patients with untreated essential hypertension?]. AB - The essential arterial hypertension is the second (after diabetes mellitus) cause of chronic renal failure which means a great social and economic burden to the society. It is well known that hypertension is a metabolic syndrome resulting in tissue injury. We tried to investigate the possible influence of some metabolic disturbances on renal function in nontreated essential hypertension. We have compared 25 patients with nontreated essential hypertension (11 women, 14 men) with 14 healthy volunteers (7 women, 7 men) matched for age. The patients' group was characterized by significantly higher urine excretion of NAG (N-acetyl-beta-D glucosaminidase) (2.75 +/- 1.69 vs 1.82 +/- 1.46 p < 0.05) and a tendency to significantly higher urine fractional sodium excretion without significant difference in albumin excretion. These findings suggest that the tubular damage is present. We noticed the negative linear correlation between mean arterial pressure and (MAP) and NAG urine excretion in the group of hypertensive patients which may reflect the renal ischemia in tubulo-interstitial pathology. Our data suggests that in nontreated arterial hypertension the renal blood flow disturbances are the important cause of the deterioration of tubular function (which are earlier to glomerular damage). PMID- 11392163 TI - [The effect of cigarette smoking and alcohol consumption on efficacy of Helicobacter pylori eradication]. AB - None of the drug regimens used to treat H. pylori infection ensures 100% of efficacy. One may think therefore that among factors modifying H. pylori eradication there are also cigarette smoking and alcohol consumption. The aim of the study was to determine the influence of cigarette smoking and alcohol consumption on efficacy of H. pylori eradication. The study was conducted on 142 H. pylori positive peptic ulcer patients treated with either OAT-omeprazole (2 x 20 mg), amoxycillin (2 x 1000 mg), tinidazole (2 x 500 mg) (69 patients) or OAC omeprazole (2 x 20 mg), amoxycillin (2 x 1000 mg), clarithromycin (2 x 250 mg) (73 patients). Detailed information on smoking and drinking habits was obtained from a questionnaire fulfilled by all subjects. Patients were defined as smokers if smoked 5 or more cigarettes per day and as drinkers if drank 25 g or more alcohol per week. To enter the study patients had to have confirmed H. pylori infection by two tests (CLO-test and histology). Eradication was considered successful if both tests gave negative results 4-6 weeks after the cessation of treatment. The efficacy of H. pylori eradication was similar in both groups (OAT- 69.6%, OAC--78.1%). Patients who smoked cigarettes had lower rate of H. pylori eradication in OAC group (smokers 65.8%, non-smokers 91.4%, p < 0.01), while patients who drank alcohol had higher eradication rate in OAT group (drinkers 85.2%, non-drinkers 59.5%, p < 0.05). When two factors (smoking, drinking) were analyzed together, it was found that in drinkers treated with OAT, smoking did not change the efficacy of H. pylori eradication, while in non-drinkers decreased by two times (75.0% vs 38.9%, p < 0.02). In drinkers treated with OAC, smoking did not change the efficacy of H. pylori eradication (but this was likely related to the limited number of patients), while decreased it in non-drinkers from 90.0% to 65.2% (p < 0.05). When two groups were analyzed together (OAT + OAC), the lowest efficacy of H. pylori eradication exhibited smokers who do not drink (53.7%) followed by smokers who drink (75.8%), non-smokers who do not drink (83.3%) and non-smokers who drink (92.9%); in each case the efficacy of eradication was higher than in smokers who do not drink (p < 0.05, p < 0.01, p < 0.01, respectively). Both cigarette smoking and alcohol consumption can affect the efficacy of H. pylori eradication. Smoking and drinking habits should be taken into account, when the set of drugs for H. pylori eradication is chosen. PMID- 11392164 TI - [Evaluation of reproducibility of plethysmographic indices according to body posture, choice of forearm and method of measurement]. AB - The aim of the study was the analysis of the influence: a) body position (sitting vs. supine), b) choice of the arm (dominant vs. nondominant), c) variant of the method (classic vs. automatic) on plethysmographic indices describing forearm blood flow repeatability (arterial inflow--AI, fast blood flow--FBF, venous outflow VO, venous capacitance--VC). The study group included subjective healthy men, aged from 22 to 60 years. Individuals with body mass index (BMI) > 35 kg/m2 or treated pharmacologically weren't been included into examination. The indices of the forearm blood flow were calculated from the plethysmographic curve using the graphic technique: venous capacitance (VC), arterial inflow (AI), fast blood flow (FBF) and venous outflow (VO). We used relative repeatability coefficient (RRC) and coefficient of variation (CV) for assessing repeatability of obtained parameters. Repeatability of plethysmographic indices was better in supine position than in sitting one. RRC was respectively for supine vs. sitting position for VO: 0.41 vs. 0.68, for VC: 0.42 vs. 0.52 and for AI: 0.57 vs. 0.65. Plethysmographic indices VO, VC, AI were characterised by better repeatability when the exams were performed on the dominant arm. RRC was respectively for dominant vs. nondominant arm for VO: 0.68 vs. 0.71, for VC: 0.52 vs. 0.64 and for AI: 0.65 vs. 0.71. Coefficient of variation of arterial inflow assessed by conventional, automatic and by fast inflow was respectively 20%, 23% and 17%. The long-term repeatability of FBF estimated by RRC was 0.76 whereas CV yielded 17%. The same coefficient of variation was obtained when short-term repeatability was estimated-mean value CV was 17%, after rejection extremal values 11%. In conclusion the best repeatability was obtained when measurements were performed with automatic variant of method, in supine position, on dominant arm. PMID- 11392165 TI - [Evaluation of thyroid structure and function in patients with Turner syndromes]. AB - The aim of our study was an estimation of thyroid structure and function in 37 patients with Turner syndrome aged from 19 to 60 years and in control group of healthy women. In each case the following studies were performed: cytogenetic examination, thyroid ultrasonography, serum total and free thyroid hormones, TSH, thyroglobulin (Tg), thyroid hormones binding globulin (TBG), antithyroglobulin and antithyroperoxidase antibodies (anti-Tg and anti-TPO) levels. In Turner syndrome ultrasonographic volume of the thyroid was significantly lower than in control group (11.03 vs 16.98 cm3). Abnormalities of thyroid function were found in 8 (22%) studied cases (subclinical primary hypothyroidism in 16%, full clinical primary hypothyroidism in 3% and hyperthyroidism in course of Graves disease in 3%). Serum elevated antithyroid antibodies were present in 62% cases of Turner syndrome and were significantly higher than in control group (16%). In Turner syndrome thyroid diseases are more frequent than in healthy population. Every patient with Turner syndrome needs routine diagnostics of the thyroid structure and function. PMID- 11392166 TI - [Familial LCAT deficiency--clinical picture. Case report]. AB - We report a case of familial LCAT deficiency, diagnosed in a 35 year old women. The disease manifested itself by a presence of proteinuria, corneal opacities and haemolytic anaemia with target cells. Suspecion of familial LCAT deficiency was based on renal biopsy, which revealed characteristic serpiginous fibrillar deposits in electron microscopy. The diagnosis was confirmed by a marked decrease of estrified cholesterol, low HDL-cholesterol concentration, decrease of LCAT activity in serum, typical "stacked coins" picture of HDL lipoproteins in electron microscopy examination and positive familial history--diagnosis of LCAT deficiency in dialysed brother of patient. PMID- 11392168 TI - [Pathogenesis of liver fibrosis]. PMID- 11392167 TI - [Is it worthwhile to treat stable angina pectoris with long-acting nitrates?]. AB - Organic nitrates have been used in the treatment of angina pectoris for over a century. During the last twenty years, their use has also been expanded to include the treatment of unstable angina and congestive heart failure. Although nitrates are among the most often used drugs in coronary artery disease, to date, their long-term use has not been investigated in any large-scale clinical trials. The finding, recently published by Nakamura et al., that long-term nitrate use may be deleterious in ischemic heart disease, may therefore be the start of a critical re-examination of this traditional form of therapy. This paper examines the different effects of nitrates in various clinical situations. The pharmacological and physiological beneficial mechanisms of nitrates in coronary artery disease, and also the potentially harmful mechanisms are discussed in the final section of this article. PMID- 11392169 TI - [Intravascular malignant lymphoma (angiotrophic large-cell lymphoma)]. AB - Intravascular lymphoma (IVL) is characterized by proliferation of large malignant lymphoid cells within the lumen of small vessels. Sites usually affected include the central nervous system and skin although involvement of multiple organ symptoms have been described. IVL is very rare and aggressive type of lymphoma. Based on review of the literature we present clinicopathological, immunohistochemical and molecular features of the IVL. The etiological possibilities are discussed. PMID- 11392170 TI - [Regulation of calcium metabolism. New pathophysiologic aspects]. PMID- 11392171 TI - Methodological and motivational factors affecting the surveillance of adverse events in hospitalized patients. AB - This is a critical review of the methodological and motivational aspects of two experiences of adverse event (AE) monitoring using a surveillance diary incorporated into the progress notes of medical records. Methodological aspects of the study were that (1) AE monitoring was to become an everyday activity of ward physicians; (2) the physicians were to express their opinions concerning the causality of each AE, which was to allow the systematic detection of adverse drug reactions and unexplained AEs; (3) the surveillance diary was to include all AEs and all prescriptions, thus allowing prescription-event monitoring; (4) the time trends of unexplained AEs were to be monitored using control cards. The experiences were discontinued because the necessary investments were attributed to other competing priorities. It follows that it is essential to develop an awareness that AE surveillance requires continuous involvement insofar as it is an essential element of quality of care. PMID- 11392172 TI - Quality management as a learning system. PMID- 11392173 TI - Reducing medication errors. AB - This article describes initiatives one institution developed to improve systems for detecting and preventing adverse medication events. Our discussion focuses on issues regarding the frequency and incidence of medication errors, the trials of traditional versus anonymous incident reporting, and the efforts to improve systems rather than placing blame and punishment on individuals. Initiatives such as improved documentation of pediatric patient weights and hepatic and renal function, increase of direct physician order entry into our Medical Information System (MIS), elimination of nonemergent verbal orders, and new and improved MIS ordering matrices (incorporating medical protocols and pathways) have led to more rational and efficient practices. Improved error prevention and critical incident review have identified on-going opportunities for improvement. Although the direct impact on patient outcomes is not yet measurable, numerous positive results have allowed for improved clinical decision making, streamlining of processes, increased regulatory compliance, and a positive culture change. PMID- 11392174 TI - Preoperative correlates of the cost of coronary artery bypass graft surgery: comparison of results from three hospitals. AB - This article furthers our understanding of the cost of coronary artery bypass graft (CABG) surgery by analyzing the extent to which preoperative correlates of cost differ among hospitals. A total of 2828 patient who underwent bypass surgery at 3 hospitals (2 teaching and 1 nonteaching) were analyzed. The preoperative correlates of direct variable cost (marginal cost) were determined by ordinary least squares regression. Age, urgent/emergent surgical priority, previous CABG, and chronic obstructive pulmonary disease (COPD) were significant contributors (P < .05) to cost in all hospitals, but overall, there were many differences. The major contributor to cost was non-white race (31.3%) at teaching hospital A, previous CABG (30.5%) at teaching hospital B, and preop insertion of intra-aortic balloon pump (IABP) (35.9%) at the nonteaching hospital. The number of significant risk factors also differed. Preoperative characteristics that contribute to cost can be quite different among hospitals and therefore results from one hospital cannot be broadly generalized to others. PMID- 11392175 TI - Improving glycemic control: can techniques used in a managed care setting be successfully adapted to a rural fee-for-service practice? AB - The objective of this work was to improve glycemic control using case management supported by electronic diabetes care monitoring. Information for patients with diagnosed diabetes in a rural community was maintained in the Diabetes Care Monitoring System. In September 1998, counseling and medication management for glycemic control was intensified during individual office visits. And, from September 1998 to February 1999, 2-hour cluster visits modeled after a successful urban program were offered for groups of patients with elevated HbA1c values. The median (and 75th percentile) HbA1c values for the patient population decreased from 8.7% (10.9%) in March 1998 (N = 173) to 7.5% (9.3%) in March 1999 (N = 182) and was maintained at 7.5% (9.1%) through March 2000 (N = 182). Case management, including cluster visits, can be accomplished in a rural physician's office with the support of an electronic diabetes care monitoring system. This intensified approach decreased and sustained the HbA1c level by more than a percentage point for the patient population. PMID- 11392176 TI - Health services utilization after induced abortions in Ontario: a comparison between community clinics and hospitals. AB - The purpose of this study was to compare postabortion health services utilization of hospital abortion patients with community clinic abortion patients using administrative databases. The study was a retrospective cohort study. The study group consisted of patients with induced abortions (n = 41,039) performed in hospitals or community clinics recorded in the 1995 Ontario Health Insurance Plan claims (OHIP) database. An age-matched cohort of 39,220 women who did not undergo induced abortions was selected from the same data source to serve as controls. The main outcome measures were health services utilization indicators constructed from OHIP data within 3 months postabortion from office consultations, emergency room consultations, and hospital admissions. Hospitalization indicators were constructed from Canadian Institute for Health Information hospital discharge data within 3 months postabortion and included data on hospitalizations for infection, certain surgical events, or psychiatric problems. Postabortion health services utilization and hospitalization were higher in the patient population, regardless of service location, than in the age-matched cohort. Within the abortion patient population, hospital day-surgery patients had higher rates of postabortion utilization and hospitalization than did community clinic patients. Multivariate analysis revealed that hospital day surgery patients had a higher risk of subsequent post-abortion hospitalizations for infections (odds ratio [OR] 1.67, 95% confidence interval [CI] 1.23-2.28), surgical events (OR 1.70, 95% CI 1.30-3.24) and psychiatric problems (OR 2.65, 95% CI 1.77-3.98) than community clinic patients. The rates of postabortion health services utilization and risk of hospitalization were lower in community clinic abortion patients than in hospital day-surgery patients. However, it is not possible to fully control for important confounding variables when using these administrative data. PMID- 11392177 TI - A look into the future. PMID- 11392178 TI - Acute tacrolimus overdose and treatment with phenytoin in liver transplant recipients. AB - As a potent immunosuppressive agent, Tacrolimus is widely used in organ transplantation. Although complications due to chronic Tacrolimus use are rather well recognized, little is known about acute overdose and its treatment. Phenytoin, an anti-convulsant agent, can induce the Cytochrome P450 enzyme in the liver, which metabolizes Tacrolimus. Therefore, it can be considered as a potential treatment option for acute Tacrolimus toxicity. We hereby report two cases of acute Tacrolimus overdose after Orthotopic Liver Transplantation and the treatment with Phenytoin. Probable mechanisms of metabolism and interactions of these two drugs are discussed and the literature is reviewed. PMID- 11392179 TI - The OU College of Medicine responds to the demand for educating medical professionals in palliative care. AB - This past decade the medical community saw an increase of national interest in the training and educating of physicians to provide quality end-of-life care for patients. This article describes the efforts of the University of Oklahoma College of Medicine to respond to the demand for educating medical professionals in end-of-life care. A Palliative Care Program was created to develop and implement new courses, seminars, and lectures for medical students, residents, and practicing physicians. Palliative medicine is in the process of being integrated into the OU academic medical environment so that all trainees, regardless of their educational level, have the opportunity for didactic and clinical exposure to end-of-life care. PMID- 11392180 TI - Public health explores expanding newborn screening for cystic fibrosis, congenital adrenal hyperplasia, and medium-chain acyl coenzyme A dehydrogenase deficiency (MCAD). AB - Advances in technology provide new challenges to public health to implement screening programs that are effective, cost-efficient, and available to all infants regardless of ability to pay. The Newborn Metabolic Disorder Screening Program (NMDSP) of the Oklahoma State Department of Health is evaluating the expansion of newborn screening for the disorders of cystic fibrosis, congenital adrenal hyperplasia, and medium-chain acyl coenzyme A dehydrogenase deficiency (MCAD) to the current screening battery of disorders (phenylketonuria, congenital hypothyroidism, galactosemia, and sickle cell disease). The challenge is to offer these new screening tests in a cost-efficient manner that ensures all newborns have access to screening and that an infrastructure exists to diagnose and provide the specialized comprehensive care affected infants will require to reduce the morbidity, mortality, and disability associated with these disorders. Essential components of an effective newborn screening system include the smooth integration of sample collection, laboratory testing, follow-up, diagnosis, timely treatment, and tracking components. The NMDSP has recommended that screening should be expanded, but issues of cost and the establishment of a sustainable infrastructure of comprehensive medical services must be addressed. PMID- 11392181 TI - PBR, MERFA among top medicine issues in Congress. PMID- 11392182 TI - Deaths among women due to lung cancer on the rise in Oklahoma. PMID- 11392183 TI - Bush administration takes action on privacy rule. PMID- 11392184 TI - Drug-seeking behavior--a differential approach. PMID- 11392187 TI - Coronary Artery Risk Detection in Appalachian Communities (CARDIAC): preliminary findings. AB - West Virginia's mortality and morbidity from cardiovascular disease (CVD) is among the highest in the developed world. Appalachia, and West Virginia in particular, could reduce the high prevalence of premature coronary heart disease (CHD) by addressing modifiable independent risk factors such as poor nutrition, sedentary lifestyle, and tobacco use. School-based health promotion programs have been shown to be an effective means of influencing student and parental health behavior. The pilot phase of the Coronary Artery Risk Detection in Appalachian Communities (CARDIAC) project substantiated an alarmingly high incidence of CVD risk factors among 347 fifth grade children from three rural counties, and was also an effective means of identifying parents at risk of developing CHD. Utilization of the innovative Rural Health Education Partnerships (WVRHEP), coupled with a health conscious public school system, offers a unique opportunity to establish the first statewide cardiovascular disease community intervention project in the nation. PMID- 11392188 TI - Blunt ureteral injury with associated aortic dissection: a case report. AB - Ureteral injury from blunt abdominal trauma is very uncommon and is reported most often in children. We report a case mid-ureteral transection following blunt abdominal trauma that resulted in nephrectomy. There was also an associated aortic dissection. The pathophysiology, epidemiology, diagnosis, and treatment of blunt ureteral injury are also discussed. PMID- 11392189 TI - Directional coronary atherectomy in acute myocardial infarction. AB - Catheter-based intervention with directional coronary atherectomy (DCA) involving the surgical removal of obstructive plaque has been extensively studied in patients exhibiting stable and unstable angina. However, since no studies to date have evaluated the effectiveness of stand alone DCA as a primary treatment in the acute myocardial infarction (MI) setting, we conducted a five-year retrospective study of the early clinical and in-hospital outcomes of patients who underwent DCA within the 72-hour interval following acute MI. Our study included data obtained from a total of 30 acute MI patients treated between 1993 and 1998. Twenty-six of these patients (86%) met the criteria for angiographic success, while 11 (36%) showed less than 50% residual stenosis without the need for supplemental percutaneous transluminal coronary angioplasty (PTCA) or additional treatment for other major complications. Four (13%) of the patients exhibited negative outcomes including post-procedural MI, emergency coronary artery bypass grafting, abrupt vessel closure, hemorrhage necessitating vessel repair and transfusion, and death. Although stand alone DCA is not presently a common treatment, our results suggest it to be an effective and relatively safe procedure for acute MI. However, further studies involving a larger number of patients and a longer follow-up evaluation are needed for comparison of the relative effectiveness of this procedure with other treatments of acute MI. PMID- 11392190 TI - Functional health literacy in adults in a rural community health center. AB - At a rural community health center, 183 adult patients were invited to participate in an unspecified study. In a closed-door session, interested invitees were told the study required taking the Test of Functional Health Literacy in Adults (TOFHLA). After the purpose of the study was privately disclosed, 70 patients agreed to enroll (38.25 percent acceptance rate). Approximately 15 percent demonstrated literacy/numeracy deficits, scoring in the Inadequate and Marginal Functional Health Literacy ranges. Student's t-test comparison of mean scores suggested patients with literacy/numeracy deficits tended to have higher mean ages and completed fewer mean years of school than those scoring in the Adequate range, p = 0.0007 and p = 0.0005, respectively. Pearson's Product Moment Correlations demonstrated a negative correlation between age and TOFHLA score, r = -0.52, a positive correlation between highest year of schooling and TOFHLA score, r = 0.51, and a negative correlation between age and highest grade of schooling, r = 0.39. Regression analysis suggested 27 percent of the variability in TOFHLA score was attributable to age, p = 0.0001, supporting the role of age in cognitive decline. Upon review of these patients' medical records, no information on their literacy/numeracy problems or deficits was identified. PMID- 11392191 TI - The counseling role of Baptist ministers in rural and urban West Virginia counties. AB - It is well established that about half of Americans suffering with a broad range of psychiatric diseases seek ministerial counseling instead of help from the medical community. Less well established is the influence of demographics, such as population density, on the mental health work of ministers. This study compares the counseling activities of Baptist ministers in rural and urban counties of West Virginia. Ministers responded to a questionnaire that assessed basic demographic data as well as attitudes regarding mental illness and counseling activity. County population density was then used to rank respondents. The respondents, who were almost exclusively male, were middle-aged and averaged greater than 20 years of experience in the ministry. They had an average church size of over 200 people. Although greater than 70% reported recent training in ministerial counseling, 85% indicated interest in a workshop focusing on common mental illnesses. No minister reported charging fees for their counseling. Significantly more counseling was reported by ministers from more urban areas with higher per capita incomes and larger congregations. PMID- 11392193 TI - The scope of the physician assistant outlined. PMID- 11392192 TI - Competencies and concerns in end-of-life care for medical students and residents. AB - A palliative medicine comfort-confidence survey developed by Weisman et al was utilized to assess self-reported competence and comfort with four end-of-life dimensions and determine future educational preferences. The survey was completed by 3rd and 4th year medical students (M3 and 4) and residents PGY 1-5. Self reported competence increased with level of training. All trainees indicated the least comfort with 1) discussing home-hospice referrals; 2) conducting a family conference; 3) discussing the change from curative to comfort care; and 4) discontinuing i.v. hydration in a dying patient not taking oral nourishment. When withdrawing parenteral antibiotics from a non-decisional dementia patient with sepsis at the request of his legal guardian, M3 and 4 expressed a greater concern for violating medical practice standards while PGY4 and 5 showed a greater concern for the violation of personal religious or ethical beliefs. Pain management, pain assessment, hospice care and end-of-life communication where the most requested topics for future education. PMID- 11392194 TI - Shakin up da joint. PMID- 11392195 TI - Thoracic aorta tear repair. PMID- 11392196 TI - Penman Lecture. The place of guidelines in surgical practice. PMID- 11392197 TI - Splenic abscess--a changing trend in treatment. AB - Fifteen patients with splenic abscesses were evaluated between 1985 and 1995. The most common predisposing factors were remote infection, diabetes mellitus and heart disease. Common clinical presentations included leucocytosis, fever, left upper quadrant abdominal pain and left pleural effusion. Four patients with splenic abscesses smaller than 4 cm in diameter were treated with antibiotics alone, and 1 in this treatment group died. Among the 10 patients with splenic abscesses larger than 4 cm in diameter receiving percutaneous drainage, 9 (90%) were successfully cured, including 8 with unilocular abscesses and 1 with multilocular abscesses. Two patients underwent splenectomy. In conclusion, percutaneous drainage using ultrasound or computed tomography guidance may be recommended as the treatment of choice for splenic abscess larger than 4 cm in diameter. Antibiotics alone may sometimes be considered for splenic abscesses smaller than 4 cm in diameter. Splenectomy is reserved for those cases where medical treatment has failed. PMID- 11392198 TI - Solid and cystic papillary epithelial neoplasm of the pancreas. A report of 3 cases. AB - This is a report of 3 female patients presenting with solid and cystic papillary epithelial neoplasms of the pancreas (SCPEN). All 3 lesions were incidental findings. SCPEN is an uncommon low-grade malignant tumour that is histologically distinct from ductal adenocarcinoma and islet cell tumour, occurs chiefly in young women, and is amenable to surgery. The cell of origin is the subject of some debate. PMID- 11392199 TI - Meckel's diverticulum duplication. Case report. AB - Meckel's diverticulum is the most common congenital abnormality of the small intestine. A case of Meckel's diverticulum duplication is described, believed to be the first report of this kind in the literature. PMID- 11392200 TI - Tonsillectomy and tonsillitis in Cape Town--age and sex of patients. AB - OBJECTIVES: To assess the sex and age of patients undergoing tonsillectomy in the Cape Town metropolitan area. METHODOLOGY: A retrospective audit was conducted from 1991 to 1995 comparing the sex and age of patients undergoing tonsillectomy with patients admitted to hospital for acute tonsillitis and the general population. RESULTS: A total of 7,250 tonsillectomies were performed for recurrent tonsillitis, and 111 adult patients were admitted with acute tonsillitis. When the data for the adult tonsillectomy patients were isolated, the female/male ratio was 3.22:1, which is highly significant when compared with the data for the general population (Cornfield's 95% confidence limit: 2.82, 3.68) and for patients with acute tonsillitis (Cornfield's 95% confidence limit: 1.14, 2.65). CONCLUSION: Recurrent tonsillitis requiring tonsillectomy occurs more commonly in female than male patients. PMID- 11392201 TI - The proper use of the microbiology laboratory guidelines and rejection criteria for common tests. PMID- 11392202 TI - Vaccination myths and misconceptions. PMID- 11392203 TI - Fallibility and risk in medicine. PMID- 11392204 TI - Beta-blockers underutilized. PMID- 11392205 TI - A practical approach to fibromyalgia. AB - The term fibromyalgia refers to a collection of symptoms with no clear physiologic cause, but the symptoms together constitute a clearly recognizable and distinct pathologic entity. The diagnosis is made through the examiner's clinical observations. The differential diagnosis must include other somatic syndromes as well as disease entities, including hepatitis, hypothyroidism, diabetes mellitus, electrolyte imbalance, multiple sclerosis, and cancer. Diagnostic criteria serve as guidelines for diagnosis, not as absolute requirements. Treatment of fibromyalgia, which is an ongoing process, remains individualized, relying on a good physician-patient relationship. It is goal oriented, directed at helping patients get restorative sleep, alleviating the somatic pains, keeping patients productive, and regulating schedules. It can be achieved through a goal-oriented agreement between patient and provider. Because fibromyalgia is chronic and may affect all areas of an individual's functioning, the physician needs to also evaluate the social support systems of patients with fibromyalgia. The approach to treatment should integrate patient education as well as non-pharmacologic and pharmacologic modalities. To keep patients well educated and involved in their healthcare, physicians should provide patients with adequate sources for reliable information. PMID- 11392206 TI - Osteopathic treatment considerations for rheumatic diseases. AB - Patients who receive medical care for musculoskeletal pain of rheumatic diseases often benefit from additional osteopathic manipulative treatment. This article offers a brief description of commonly used treatment modalities. It also includes discussion of indications as well as contraindications of operator director versus operator-monitored techniques. PMID- 11392207 TI - Osteoarthritis. AB - Arthritis is a common, disabling condition that affects an increasing percentage of the population. This article includes discussion of new theories of risk factors and treatment paradigms. PMID- 11392208 TI - A nonsurgical approach to low back pain. AB - Low back pain, a leading cause of disability in the United States, has a significant economic impact not only on lost productivity but also on healthcare expenditures. Approximately a fifth of patients will see multiple physicians in their quest for relief of low back pain. Primary care physicians therefore play a crucial role in the initial approach to these patients. A thorough history and physical examination directed toward the neurologic, orthopedic, and osteopathic evaluation are essential. This article reviews the diagnosis and assessment of pain levels and a triad system of therapy involving cortical, spinal, and peripheral levels. Options include antidepressants, neuroleptics, neurostimulants, and osteopathic manipulative treatment (OMT) (cortical level); opiates, tramadol hydrochloride, and transcutaneous electrical nerve stimulators (spinal level); and nonsteroidal anti-inflammatory drugs, epidural injections, spinal blocks, antispasmodics, physical therapy, muscle relaxants, exercise, and OMT (peripheral level), By choosing a modality directed at each level, the clinician may provide the patient with a pain management program that will maximize the chosen mode of therapy and restore function and mobility. PMID- 11392209 TI - Arthritis is a term used to describe more than 100 different rheumatic diseases. PMID- 11392210 TI - Irritable bowel syndrome. AB - Irritable bowel syndrome is one of the most commonly encountered functional gastrointestinal disorders, affecting an estimated one in five adult Americans. It is characterized by a symptom complex that includes chronic abdominal pain and discomfort that is relieved with defecation, abnormal stool frequency, and a change in stool form. Although the etiology of this disorder continues to remain elusive, this article reviews the current theory of the pathophysiologic mechanisms and reports on diagnostic and management principles for the primary care provider. PMID- 11392211 TI - Lactose intolerance. AB - Lactose intolerance affects more than 50 million Americans. It is one of the most common gastrointestinal disorders seen by primary care physicians. When this disorder is properly diagnosed, the patient is easily treated with education and dietary modifications. Lactose intolerance is commonly misdiagnosed because of its overlapping symptoms of diarrhea and abdominal bloating. This article reviews the etiology, diagnosis, and treatment of lactose intolerance. PMID- 11392212 TI - Enteral and parenteral nutrition support. AB - The use of enteral and parenteral nutrition support may be warranted when patients are unable to ingest adequate calories or protein (or both). This review provides an overview of the indications, access, administration, management, and complications of these modes of therapy. PMID- 11392213 TI - Overview and treatment of diverticular disease. AB - Diverticular disease is a common medical problem seen in Western society. Outpatient management with close observation is appropriate for the majority of patients. Established criteria for hospitalization and treatment of diverticulitis can help to reduce medical costs and length of stay. Minimally invasive techniques such as computed tomography-guided drainage of diverticular abscess can expedite medical and surgical treatment. PMID- 11392214 TI - Acute pancreatitis. AB - Acute pancreatitis is a relatively common disease with an incidence of 30 per 100,000 persons per year. In the United States, more than 80% of the cases are related to biliary stones or alcohol use. It is a potentially fatal disease with an overall mortality of 5% to 10%. When first seen, patients have an acute abdomen. It is imperative that the diagnosis be established rapidly with a thorough history, physical examination, and appropriate laboratory and imaging studies. Prompt determination of patients who need referral for intensive care or consultation is crucial. There is no specific treatment for most patients with acute pancreatitis. Supportive care includes intravenous administration of fluids, parenterally administered analgesia, nutritional support, and prevention and treatment of complications. PMID- 11392215 TI - [Malignant tumors in patients with kidney transplants ]. AB - In this retrospective study we evaluated incidence of malignancies observed among 718 renal transplant recipients with at least 6 months of follow-up. A total of 461 men and 257 women (mean age at transplantation 36.3 +/- 8.3 years) were included. Thirty three out of 718 recipients (4.6%) developed malignant neoplasia: 45.4% of these were Kaposi's sarcomas, 12.1%--cancers of the uterine cervix, 12.1% cancers of the stomach, 12.1%--basal cell carcinomas, 6.06%- posttransplant lymphoproliferative disorder. There was no significant effect of either cyclosporin A doses or OKT3/ATG on the incidence of the tumors. Mean age of transplant recipients with malignancies was statistically higher as compared to those without malignancies (45.5 +/- 8.2 years versus 36.1 +/- 8.4 years, p < 0.00001). The median time from onset of end-stage renal failure (dialysis start) and from the transplantation to the diagnosis of the tumor make up 32 (16-161) and 23 (5-158) months, respectively. One renal transplant patient suffered from multiple myeloma with aggressive course. PMID- 11392216 TI - [Treatment of benign prostatic hyperplasia with the preparation Gentos]. PMID- 11392217 TI - [Tamsulosin in the treatment of patients with chronic prostatitis]. AB - Combined treatment including alpha-1a-adrenoreceptor blocker tamsulosine was given to 92 patients with chronic prostatitis having marked urination disorders (IPSS score 12.1 +/- 2.4). The response was achieved in 98.9% of cases. There was attenuation of the urination symptoms, speeding up of the maximal urine flow, decreased amount of the residual urine. It is recommended to add alpha-1 adrenoreceptors to combined treatment of chronic prostatitis patients with urination disorders. PMID- 11392218 TI - [Treatment of patients with chronic prostatitis]. PMID- 11392219 TI - [Comparative effectiveness of the strategies of etiotropic therapy of generalized chlamydia infections in young patients]. PMID- 11392220 TI - [Perspectives of introducing a system of sanatorium-spa rehabilitation of patients with chronic renal failure]. PMID- 11392221 TI - [Bladder wall permeability to to topically administered cisplatin]. AB - The aim of the study was quantitation of cisplatinum in the wall of normal urinary bladder, tumor tissue, inflamed mucosa and blood in intravesical administration of cisplatinum. The samples of the tissue were radiated in the flow of heat neutrons 5 x 10(12)-5 x 10(13) neut/cm followed by radiochemical purification of the material and platinum assay in the tissue using analyser LP 4900 with semiconductor radiation detector. The samples were taken from 32 patients with transitional cancer of the bladder. Tumor tissue contained platinum in amounts 33.7 times exceeding those in normal tissue after intravesical administration of 100 mg of the drug. After intravenous administration of 100 mg cisplatinum normal bladder tissue and tumor tissue contained almost similar quantities of the drug. Thus, tumor tissue absorbs more cisplatinum than normal tissue of the bladder in intravesical administration. PMID- 11392222 TI - [Initial results of the clinical use of a novel Russian lithotriptor LitURAT-UR]. AB - By its performance characteristics LitURAT-UR lithotriptor is different from previous units made in Russia or abroad. Besides the double stone-directing system (ultrasound + x-ray), LitURAT-UR is furnished with two interchangeable generators of shock-wave impulses (electrohydraulic and electromagnetic) allowing effective destruction of any urolith. The TV system is able to control the interelectrode space and to change it in the course of lithotripsy. Efficiency of the new lithotriptor in different clinical forms of urolithiasis reached 93.1%. Thus, universal, effective and low-traumatic characteristics of the lithotriptor LitURAT-UR hold much promise for urological practice in Russia. PMID- 11392223 TI - [Objective control over the degree of tension of the pubic-vaginal ligatures in transcutaneous retropubic urethrocervicopexy in women with stress urinary incontinence]. PMID- 11392224 TI - [Long-term results of the surgical treatment of primary obstructive megaureter in children]. AB - In 1989-1998 reconstructive plastic operations for primary obstructive megaureter (POM) were made in 71 children. Ureterocystoneostomy with antireflux defense was made in 35 children. Prior trocar nephrostomy was performed in 5 patients with bilateral megaureter and decompensation of the upper urinary tracts. Extensive resections of the dilated ureters with their longitudinal modelling were not used. Long-term results were good: 85.7% of the children recovered, 8.6% improved. Morphometric examinations of the distal obturating ureters before surgery and 6 months to 9 years after it showed that after operative reestablishment of ureteral patency diameter of the ureter reduced 6.2- and 5.5 fold, on the average, in children under 5 years and 6-14-year-olds, respectively. It was also demonstrated that recovery of ureteral contractility and diminution of its size need time (at least 1-1.5 years). The author does not recommend to make longitudinal modelling of the ureter in children, especially in infants. PMID- 11392225 TI - [Radiographic-endoscopic diagnosis and treatment of obliterations of the upper urinary tract]. AB - Potentialities of x-ray endoscopic diagnosis and treatment of upper urinary tracts (UUT) obliterations were studied in 26 and 21 patients, respectively. The causes of obliteration were previous urological, surgical, gynecological operations, radiotherapy (one case). Percutaneous or transurethral UUT recanalization was performed in 5 and 16 patients, respectively. The length of the obliteration varied from 0.3 to 1.7 cm. It was located at the level of ureteropelvic segment in 13(62%), in the upper third of the ureter in 4(19%) and in the lower third of the ureter in 4(19%) patients. UUT endoscopic recanalization was successfully performed in 17(81%) patients. The operation took 60-90 minutes. Minor complications were corrected conservatively. Control examination was made after 6 to 15 years follow-up. The result was assessed as positive in the absence of clinical symptoms, recurrent stricture(obliteration), in improvement or no change in renal function. These criteria were met in 14(66.7%) patients. One patient has undergone nephrectomy because of highly deficient renal function and dislocation of the nitinol spiral outside recanalization zone. Two patients live with drained kidney in spite of UUT restored patency. The method proved effective and can be used as first-line therapy in short obliteration (< 1 cm), relatively intact renal function and in the absence of UUT marked hypotonia, more so as the failure does not complicate further surgery. Combined x-ray endoscopic diagnosis is presently most significant in determination of complete stricture(obliteration) of ureteropelvic segment and the ureter, its length and location. PMID- 11392227 TI - [Results of puncture diagnostic and therapeutic interventions on the retroperitoneal and small pelvis organs under ultrasonic control]. PMID- 11392226 TI - [ALA fluorescent diagnosis of bladder cancer]. AB - Application of 5-aminolauvulin acid (ALA) fluorescence allows to detect not only exophytic tumors of the bladder but also flat, small tumors-satellites and preneoplastic changes of the bladder. 175 biopsies were performed in 53 patients with suspected superficial tumor of the bladder. 3 hours before surgery all the patients were intravesically instilled 50 ml 3% ALA solution. Cystoscopy employed white and blue light. Visual registration of exophytic masses and red fluorescence of the suspected sites was registered and consequently compared to the histological findings. 96 of 100 sites with malignancy/dysplasia showed red fluorescence. In 13% patients cancer and mucosal dysplasia were detected only under the blue light and were missed by standard cystoscopy. Residual red fluorescence of the resection margins was observed in 41% patients after TUR. Sensitivity of ALA-fluorescent detection reached 96%, specificity 52%. ALA induced fluorescent diagnosis is more effective than standard cystoscopy. It is most effective in diagnosis of dysplasias, carcinoma in situ, flat, small, multiple superficial tumors of the bladder during primary TUR. PMID- 11392228 TI - [Laparoscopic pelvic lymphadenectomy in patients with cancer of the bladder and prostate]. PMID- 11392230 TI - [Surgical and functional therapies of disorders of muscular equilibrium]. PMID- 11392229 TI - [Laboratory methods of predicting primary and recurrent nephrolithiasis]. AB - Concentrations of Na, K, Ca ions, oxalic and uric acids, total Ca, P as well as urease activity, the ability to form crystals, speed of crystallization, crystal chemical composition, pH stability were evaluated in the urine from 40 urolithiasis patients and 40 patients with recurrent nephroliths. It was found that fast or moderate crystallization, high urease activity and ability to form crystals, unstable urine pH indicate a high risk of both primary and recurrent nephrolith formation. PMID- 11392231 TI - [Treatment of dentomaxillary disharmony]. PMID- 11392232 TI - [Pathology and treatment of the transverse dimension in the mixed dentition. Impact on muscular equilibrium]. PMID- 11392233 TI - [Pathology and treatment of the vertical dimension in the mixed dentition. Impact on muscular equilibrium]. PMID- 11392234 TI - [Pathology and treatment of the sagittal dimension in the mixed dentition. Impact on muscular equilibrium]. PMID- 11392235 TI - [Treatment during the mixed dentition and muscular equilibrium. Conclusion]. PMID- 11392236 TI - [Treatment during the mixed dentition and muscular equilibrium. Introduction]. PMID- 11392237 TI - [Maturation of muscular equilibrium in relation to the establishment of the mixed dentition]. PMID- 11392238 TI - [Relations between the mixed dentition, muscle equilibrium and biological age]. PMID- 11392239 TI - [Diagnosis of muscular imbalance]. PMID- 11392240 TI - [Clinical forms of muscular imbalance in the mixed dentition. Morphogenetic consequences]. PMID- 11392241 TI - [Limits to arterial embolization treatment of severe postpartum hemorrhage]. AB - OBJECTIVE: To assess the limits of arterial embolization in the management of serious postpartum haemorrhage (PPH). STUDY DESIGN: Retrospective study. PATIENTS AND METHODS: We examined the cases of 29 patients admitted to intensive care units of Rouen University Hospital for PPH between January 1994 and August 1999. Demographic, obstetrical and biological data, the required treatment and eventual side effects were collected and analysed using the appropriate parametric and non parametric tests. RESULTS: Arterial embolization was carried out on 15 patients (52%) with a success rate of 73%. Of the 14 other patients, 11 underwent conservative or radical surgery without further complications, three received medical treatment. No maternal death occurred; however, one patient transferred from a local hospital and already presenting haemodynamic instability suffered cardiac arrest before embolization. Arterial embolization was unsuccessful in four cases, two of which were cases of placenta accreta. These patients were older (p < 0.05) and all had a past history of curettage and/or caesarean section for preceding deliveries (p < 0.01). CONCLUSIONS: Emergency arterial embolization is a valuable therapy in case of PPH but can only be carried out in specialised units. Obstetrical antecedents would appear to constitute a major risk factor and transfer increases the morbidity rate. PMID- 11392242 TI - [Maternal serum anti-D antibody concentration monitoring and transfusion practices in obstetrics in France: a preliminary study]. AB - OBJECTIVES: In obstetrics, transfusion medicine is involved because of the risk of Rh isoimmunisation and of transfusion at delivery. In France, although these two situations have led to legal or professional recommendations, practices appear to be very different from one institution or one physician to another. STUDY DESIGN: Retrospective study. METHODS: A questionnaire using both direct questions and clinical cases was sent to all members of the French obstetric anaesthetists association (named CARO). RESULTS: One hundred and seventy-eight questionnaires (response rate 17.1%) were returned. Monitoring of anti-D antibody concentrations during pregnancy was performed in accordance with the legal recommendation (14/02/1992) in only 3.9% (Rh+) and 19% (Rh-) of women. Blood samples were performed either too often or too rarely in 39 to 72% of responses. At delivery, an immediate assessment of anti-D antibody concentration was required in almost 100% by the specialist on call in the blood bank while this measurement was considered necessary by only 80% of anaesthetists responding to the questionnaire. The distance between the maternity and the blood bank is greater than 14 km in 19% of responses and the largest time interval reported to obtain red cell concentrates was greater than 145 min in 18% of responses. Autologous blood transfusion is described as being used routinely in only 23% of responses. CONCLUSION: Significant failures in practice patterns and organisation of care are suggested by the responses to this questionnaire. This should stimulate a national debate to improve adequacy of practices to legal and professional recommendations and to review blood distribution to maternities in this country. PMID- 11392243 TI - [Pulmonary artery catheterization in the intensive care unit: at what time?]. AB - OBJECTIVES: Recording the time at which the insertion of a pulmonary artery catheter was decided. STUDY DESIGN: Prospective and descriptive study. PATIENTS: Critically ill patients in an university hospital. METHODS: The times at which the insertion of a PAC was decided were recorded. For each pulmonary artery catheterization, immediate complications were recorded (arterial puncture, pneumothorax, ventricular arrhythmia, hard and failed pulmonary artery catheterization). RESULTS: One hundred and forty-nine patients were included (99 males, age = 63 +/- 15 year, body mass index = 25 +/- 6 kg.m-2, median Apache II score = 16). One hundred and sixty-five PAC insertions were decided (16 patients requiring two PACs). Nine arterial punctures, two pneumothoraces, 42 ventricular arrhythmias, 32 hard and eight failed pulmonary artery catheterizations occurred. Thirty-four PAC insertions were decided between 9 and 10 am whereas = 3 PAC insertions per hour were decided between 1 and 9 am. CONCLUSION: The rate of decision of PAC insertion are decreased during the second half of the night (1 to 9 AM). PMID- 11392244 TI - [Immediate evaluation of continuous medical education]. AB - OBJECTIVES: To analyse the results of the immediate evaluation of an European teaching session using a questionnaire provided by the French College of anaesthesiologists. STUDY DESIGN: Open evaluation. MATERIAL: Questionnaires completed after each topic by 50 participants of an European course including 15 lectures were collected. METHODS: The types of professional exercise and the medical practise reliable to the different topics were pointed out. The evaluation included 4 items noted from 1 to 10: new information for medical practise, definition of pedagogic objectives, quality of means used for teaching, interest for the treated subject. The global mean score for each item and for each speaker was calculated. Results were compared according to the professional mode of exercise, the own medical practise and the project to modify it in the future. RESULTS: Scores affected to items were significantly lower for participants exercising in PSPH compared to general and university hospitals and private clinics (p < 0.001). New information was better found in case of poor or absence of practise, but interest was improved when medical practise was frequent. 2/3 of the participants projected to modify their practise after the session. CONCLUSION: The analysis of medical evaluation should allow to determine an acceptable zone of quality which may be useful for accreditation. However, distorting results according to the professional mode of exercise and the own medical practise should encourage the development of adapted continuous medical education. Delayed evaluation may be necessary to objective the putative benefits of CME on medical practise. PMID- 11392245 TI - [Acute pain measurement in animals. Part 1]. AB - OBJECTIVE: To describe tests of nociception which appear in the "pre-clinical" literature. DATA SOURCES: References obtained by computerized bibliographic research (Medline) and the authors' personal data. DATA SYNTHESIS: Ethical problems arising from the study of the pain in awake animals, problems arising from the choice of stimulus and stimulus parameters and the quantification of responses are presented. Pain in animals can be estimated only by examining their reactions, but at the same time, the existence of a reaction does not necessarily mean that there is a concomitant sensation. A description of the signs of pain in mammals is proposed. A noxious stimulus can be defined by its physical nature, its site of application and what has previously happened to the tissues at this site. Electrical stimulation short-circuits the process of transduction at free nerve endings and is not specific; however it has the advantage that it can be applied suddenly and briefly and thus results in synchronised signals in the relevant primary afferent fibres which can be differentiated into A delta and C fibres. Heat selectively stimulates thermoreceptors and nociceptors, but the low calorific power of conventional stimulators restricts their usefulness. Radiant sources have the disadvantage of emitting waves in the visible and the adjacent infrared spectra, for which the skin is a poor absorber and good reflector. Thermodes have the disadvantage of activating mechanoreceptors and thermoreceptors simultaneously; furthermore, their capacity for transferring heat depends on the quality of contact with skin and thus on the pressure with which they are applied. These problems can be overcome by using CO2 lasers but even today, the cost of these is a major disadvantage. Chemical stimuli differ from those mentioned above by the progressive onset of their effectiveness, their duration of action and the fact that they are of an inescapable nature. Experimental models employing chemical stimuli are undoubtedly the most similar to acute clinical pain. A wide spectrum of reactions are observed in nociceptive tests, but in almost every case they involve motor responses. After defining the ideal characteristics of a nociceptive test, tests based on the use of short duration and longer duration stimuli are presented. In tests of phasic pain, reactions are evoked by thermal (tail-flick test, hot-plate test), mechanical or electrical (flinch-jump test, vocalisation test) stimuli. Tests of tonic pain employ injections of algogenic agents intradermally (formalin test) or intraperitoneally (writhing test) or even the dilation of hollow organs. All these tests will be critically appraised in a subsequent paper [1]. CONCLUSION: The tail-flick and hot-plate tests are the most used, but there is an increasing recourse to the formalin test and tests involving foot withdrawal after mechanical stimulation. PMID- 11392246 TI - [Difficult reintubation during a Fantoni's tracheostomy resolved using a laryngeal mask]. AB - We report the failure of a reintubation under direct laryngoscopy, during a percutaneous tracheostomy by Fantoni method. After the accidental per-procedural extubation and in front of an unpredicted difficult conventional endotracheal intubation, a laryngeal mask was used as an airway intubator conduit for the blind passage of the set's ventilation tube. This ventilation tube is a 40 cm long, 4 mm internal diameter special catheter with low pressure cuff, included into the TLT kit. The end of the procedure was uneventful. PMID- 11392247 TI - [Barotrama during apnea testing for the determination of brain death]. AB - We describe three cases of tension pneumothorax occurring during apnea testing for the determination of brain death. Every case needed needle thoracostomy for emergency chest deflation and/or a chest tube to be inserted rapidly. Moreover, haemodynamic and oxygenation parameters were impaired in each of the patients after these pneumothorax. This was uneventful for the two first patients (organs harvesting was contra-indicated or not consented by the patient's family), but might be responsible for damaging lungs in the third patient and consequently loosing the pulmonary graft. Limitation of oxygen insufflation up to 8 L.min-1 with a 12 F oxygen supply tubing inserted within 5 cm into the endotracheal tube should be recommended to avoid this iatrogenic complication. PMID- 11392248 TI - [Esophageal bezoar resulting from nasogastric enteral feeding in an intensive care unit]. AB - We report two cases of oesophageal bezoar in patients given enteral nutrition by nasogastric tubing in an intensive care unit. These two complications occurred during the year following the replacement of our standard enteral feed by a new preparation enriched in proteins. In both patients, the bezoar could be endoscopically removed. The enteral feeding solution is likely to be responsible for the development of this complication because no other factor known to favour this complication such as concomitant administration of sulfacrate or anti-acid agents was given to the patients; and the bezoar developed shortly after the new enteral feeding solution was used, a hypothesis supported by several similar case reports in the medical literature. PMID- 11392249 TI - [Initial experience with a wire-guided endobronchial blockade to achieve one-lung ventilation]. AB - We report our initial experience with a wire-guided endobronchial blockade, which is a new method to achieve one-lung ventilation with a conventional endotracheal tube. The strong points of this device are its ease of use and the fast training, the possibility of setting it up after the patient was positioned in lateral decubitus position or in the course of intervention as well as the maintenance of ventilation during insertion. Its weak points are the lack of a paediatric model and the quality of the lung collapse which requires a particular operation to be perfect. PMID- 11392250 TI - [Candidiasis among non-neutropenic patients: from colonization to infection]. AB - Invasive candidiasis is a dread complication in hospitalized patients, characterized by a mortality comparable to that of septic shock (40% to 60%). Its incidence in hospitalized patients is 0.5/1000 admissions, but it complicates about 10 per 1,000 admissions in critical care where it represents 10% to 15% of all nosocomial infections. Although a high proportion of hospitalized patients may become colonized with Candida spp, the clinical signs of infection manifest only late, rending it difficult to diagnose. A better knowledge of their pathophysiology and the availability of triazoles compounds, less toxic than amphotericin B, allowed the concept of early empirical or preemptive treatment. These strategies are based on the prompt identification of risk factors and require continuous attention from skilled physicians. However, the prescription of triazoles has to be restricted to carefully selected groups of patients to avoid the emergence and the dissemination of resistant strains. PMID- 11392251 TI - [Antifungal drugs in the treatment of systemic candidiasis: susceptibility to antifungal drugs, drug resistance, pharmacological data]. AB - The available antifungal agents are amphotericin B (conventional or lipid formulation), flucytosin and azole derivatives (ketoconazole, fluconazole, itraconazole). The main target of these molecules are a specific compound of fungal membrane, ergosterol. Determination of the fungal sensitivity to antifungal drugs is difficult and no consensus has been achieved so far. Minimal inhibitory concentrations are poor predictors of clinical success or failure. A good correlation between in vitro and in vivo results has been observed only in patients with oropharyngeal candidiasis associated with HIV infection. Combinations of antifungal drugs are currently under study. The role of hemopoietic growth factors (G-CSF, GM-CSF) as an adjuvant has not been fully established. New antifungal drugs (triazole derivatives, echinocandins) should be available within months. PMID- 11392252 TI - [Epidemiology and prognostic factors of systemic candidiasis in intensive care units]. AB - The incidence of systemic Candida infections in patients requiring intensive care has substantially increased in recent years. Epidemiologic evidence for this trend is consistent in american as european studies. An emergence of non-albicans species is also demonstrated. Systemic Candida infections are associated with a high mortality and morbidity, an increased length of stay and a heavy workload for the ICU staff. A number of studies have identified risk factors for transition from mere colonisation to infection and prognostic factors for mortality. A review of published data is given. PMID- 11392253 TI - [Yeast count in urine. Review of the literature and preliminary results of a multicenter prospective study carried out in 15 hospital centers]. AB - Candida spp. are increasingly involved in nosocomial infections in severely ill patients and the diagnosis is difficult. In this context, the significance of candiduria remains unclear. Management of this condition is still equivocal, because of the lack of information about its natural history and its predictive value for disseminated infection. Little is known about the discriminant value of colony count. After a comprehensive review of the available published data, the preliminary results of a multicentric prospective survey in critically ill patients hospitalised in intensive care units are given. The aim of the study was to search for correlations between quantitative candiduria and known risk factors for disseminated candidiasis. There is a statistically significant correlation (p = 0.003) between heavy candiduria (> 10(4) cfu.mL-1) and high Pittet colonisation index (> or = 0.5). Quantification of candiduria could be useful to select patients at high risk for disseminated candidiasis. PMID- 11392254 TI - [The role of the laboratory in the diagnosis and prevention of fungal infections]. AB - Fungal infections account for an increasing fraction of nosocomial infections. The laboratory is essential to ascertain the diagnosis. Significant progress has been recently achieved in direct diagnostic tools (better sensitivity of direct examination, development of chromogenic identification media). Diagnostic sensitivity of blood cultures can be increased with the use of special fungal media for culture. However, indirect diagnostic methods remain unsatisfactory, even though some new data support the usefulness of simultaneous detection of circulating antigens and corresponding antibodies. In the future, the contribution of molecular biology techniques could be of great interest, either in the diagnostic (detection of fungal DNA in the serum) or in the identification of species. Last but not least, molecular typing allows a precise monitoring of fungal ecology that could prove useful to the understanding of the physiopathology of infections and to the anticipation and prevention of epidemic outbreaks. PMID- 11392255 TI - [Fungal infections in intensive care units: a cross-survey]. AB - STUDY DESIGN: Diagnosis and treatment of fungal infections in medical and surgical intensive care units are controversial issues. The aim of the present survey was to assess the management of fungal infections in these units in France. MATERIAL AND METHODS: Transversal study, by means of questionnaires, in a representative sample of French intensive care units, stratified by region and by status of the center. RESULTS: Eighty-two out of 704 questionnaires were returned (11.6%). The distribution was as follows: academic centers (AC), 52%; local hospitals (LH), 31%; private centers (PC), 17%. In vitro assessment of sensitivity to antifungal drugs was done "sometimes" in 43% of the centers, "always" in 35%. Serologic tests for Candida were performed in 63% of the cases. Antigenemia and PCR testing were nearly never performed. Only 64% of physicians always prescribed an antifungal drug in patients with candidemia (AC > LH = EP, p < 0.05). By contrast, in patients with candiduria and a urinary catheter, a treatment was more frequently thought necessary in LH and EP than in AC (p < 0.05). Empirical treatment was more common in public (70%) than in private (36%) centers (p < 0.05). Increasing doses of amphotericin B, without loading dose, was the generally preferred regimen (55%), and the vehicle was isotonic glucose solution (60%). The mean daily dose of fluconazole was 334 mg, after a mean loading dose of 624 mg. There was no significant differences between the centers. CONCLUSION: Management of fungal infections in intensive care patients appeared to be quite homogeneous, despite some discrepancies between centers about diagnostic procedures and specific indications for antifungal treatment. Attempts should be made to achieve a consensus in the setting of fungal infections. PMID- 11392256 TI - [Use of the colonization index]. AB - In a retrospective study, 2,238 mycologic samples obtained in 1999 from 89 patients hospitalised in an intensive care unit dedicated to digestive diseases were analysed. Feasibility of monitoring fungal colonisation and implications for workload and costs were assessed. From this experience, we confirmed the ability of the Pittet index to identify patients at high risk for Candida infection. Monitoring of Pittet index required a high degree of cooperation between the intensive care unit and the laboratory of mycology, and a precise definition of the modalities of sampling. It entailed a significant increase in costs and workload. A treatment was started whenever a colonisation index > or = 0.5 was associated with severe clinical or biological signs. This involved an increase of the expense of antifungal drugs. The potential benefits could not be evaluated from our study. Direct observation of pseudomycelium in the samples and candiduria were significantly correlated to fungal colonisation. PMID- 11392257 TI - [The rational use of aprotinin during orthotopic liver transplantation]. PMID- 11392258 TI - [Is ketamine useful for anesthesia in morbid obesity?]. PMID- 11392259 TI - [Fatal hematochezia complicating sigmoid fistulization of an aorto-iliac bypass false aneurysm]. PMID- 11392260 TI - [Transcranial doppler as a diagnostic tool for resistant hypoxemia in patent formen ovale]. PMID- 11392261 TI - [Prevention of perianesthetic allergic risk]. PMID- 11392262 TI - [Editorial on "Clinical findings with mycophenolate mofetil in immunosuppressive therapy of ocular pemphigoid" by J.Zurdel, Bilal Aboalchamat, Manfred Zierhut, Nicole Stubiger, Alexander Bialasie et al. in Klin Monatsbl Augenheilkd 2001:218:222-8]. PMID- 11392263 TI - [Effect of duration of phacoemulsification on postoperative inflammation--a retrospective study]. AB - BACKGROUND: The early postoperative inflammation after cataract surgery is mainly caused by surgical trauma. PATIENTS AND METHODS: 450 data-sheets of patients, who were operated for senile cataract with small-incision phacoemulsification and in the-bag implantation of a foldable intraocular lens were retrospectively analysed. Postoperative inflammation was evaluated with the Laser Flare-Cell Meter Kowa FC-1000 on day 1, 3, 7 and 28. All the operations were done with the same phacomachine model Orbit Oertli. The absolute phacotime was measured, the data classified in steps of 20 seconds. Statistical analysis was made with the student's t-test. RESULTS: In the first postoperative week there is a statistical significant difference in the flare-values between the groups with short phacotime (up to 39 seconds) and the group with long phacotimes (over 80 seconds). One month after operation this significance was absent. CONCLUSIONS: The early postoperative inflammation is influenced by the duration of phacoemulsification. Surgical techniques and phacomachines that may reduce phacotime are helpful with respect to early postoperative inflammation. PMID- 11392264 TI - [Transitory intrathoracic and -abdominal pressure elevation in the history of 64 patients with normal pressure glaucoma]. AB - BACKGROUND: As one pathogenetic factor in normal-tension glaucoma an individually elevated sensitivity of lamina cribrosa regarding intraocular pressure fluctuations is postulated. Aim of this study was to evaluate patients with normal-tension glaucoma for the exposure to potential, clinically undetected transient elevations of intraocular tension due to increased intrathoracic and abdominal pressure. PATIENTS AND METHODS: A survey of 64 patients of the "Erlanger Glaucoma registry" with normal-tension glaucoma (NTG) and 64 patients with primary open angle glaucoma (pOAG) as control group were performed with regard to activities respectively diseases causing intrathoracic or -abdominal pressure elevation (1. weight lifting, 2. playing high resistance wind instruments, 3. chronic asthma/cough, 4. obstruction of the urinary system, 5. constipation). Both groups were matched regarding age (median: 61 years), sex (24 male, 40 female), visual field defects (mean defect: NTG 4.4; pOAG 4.7), visual acuity (median 1.0 +/- 0.2) and systemic diseases (diabetes mellitus, hypertension, cardiac disease). RESULTS: Among patients with NTG there were 45% (29/64 patients) with activities respectively diseases causing intrathoracic or intraabdominal pressure elevation in their medical history compared to 11% (7/64) among patients with pOAG. Male patients with normal-tension glaucoma showed with 62% the highest frequency of such activities (among them 4/24 high resistance wind instrument playing, 5/24 urinary system obstructions, 4/24 long time weight lifting). Female patients with normal-tension glaucoma most frequently presented with a history of weight lifting (11/40). CONCLUSION: Patients with glaucomatous optic nerve atrophy without evident intraocular pressure elevation compared to patients with pOWG more frequently report activities or diseases causing intrathoracic/-abdominal pressure elevation in their medical history. This may suggest an additional pathomechanism in normal-tension glaucoma. Therefore patients may be adviced on these potential risk factors. PMID- 11392265 TI - [Absolute filling defects of the optic disc in fluorescein angiograms in glaucoma -a retrospective clinical study]. AB - BACKGROUND: Analysis of clinical importance of the size of filling defects in fluorescein angiograms in primary open-angle glaucoma (POAG), normal-tension glaucoma (NTG), ocular hypertension and subjects with physiological excavations in comparison to visual field loss, optic nerve head morphology and hemodynamics. PATIENTS AND METHODS: 75 patients (POAG, NTG, ocular hypertension) and 10 healthy subjects with physiological excavations were included in this study. In digitized video fluorescein angiograms (Scanning Laser Ophthalmoscope) the size of absolute filling defects of the optic disc was quantified in the early venous phase and expressed by percentage of the optic disc. Visual fields were obtained by conventional static perimetry (Humphrey 24-2) and graded in stages of glaucoma visual field defects (Aulhorn I-V). Optic disc excavations were evaluated as cup to-disc-area-ratios. RESULTS: The filling defects correlated with the visual field loss stages of Aulhorn and the visual field indices MD (mean deviation), PSD (pattern standard deviation) and CPSD (corrected pattern standard deviation). There was no correlation with the index SF (short-term fluctuation) and with systemic hemodynamics (blood pressure, perfusion pressure) or the IOP. Absolute filling defects correlated with the cup-to-disc-area-ratio in NTG. The absolute filling defects were larger in patients with glaucoma (POAG, NTG) in comparison to patients without glaucomatous visual field loss (ocular hypertension, glaucoma like discs). No difference of filling defects was found in the glaucoma group (POAG, NTG). Patients with NTG had larger excavations and lower systolic blood pressures than patients with POAG. CONCLUSION: The size of fluorescein filling defects may be useful as a parameter for the evaluation of an ischemic lesion of the optic nerve head. Absolute filling defects may differentiate POAG from ocular hypertension and NTG from glaucoma-like discs without field defects. The results support the hypothesis that in POAG and NTG disturbances of the circulation result in similar filling defects of the optic disc and visual field loss. PMID- 11392267 TI - [Eyelid and facial injuries due to dog bites]. AB - BACKGROUND: Head und neck injuries of children are mistly due to dog bite. PATIENTS AND METHODS: During the 10-year interval from 1990 to 2000 sixteen patients suffering from dog bite injuries were seen in our department. Age, gender, pattern of ocular and periocular wounds, surgical management and long term damage were analyzed. RESULTS: Medial lower eyelid injuries with lacrimal duct involvement were most common. Ten victims were younger than four years. 87% of the dogs belonged to the family and friends of the patients. Two localized infections and no systemic infection were seen. CONCLUSION: Primate microsurgical wound repair leads to a satisfying functional, morphological, and esthetic long term result. PMID- 11392266 TI - [Early clinical results with mycophenolate mofetil in immunosuppressive therapy of ocular pemphigoid]. AB - BACKGROUND: Ocular cicatrical pemphigoid can lead to severe structural damage or loss of vision at worst. Longterm therapy with dapsone or systemic immunosuppressive therapy, e.g. with cyclophosphamide is often inevitable. Immunosuppression may cause severe side effects in some patients. PATIENTS AND METHODS: Data are presented on 5 patients with ocular cicatrical pemphigoid who were treated with mycophenolate mofetil 2 g daily. Criterion of effectiveness was the clinical course of the condition defined as nonprogression of the morphologic alterations. Patients were initially examined and interviewed routinely every four weeks for the first four months, then every eight weeks. Patients were asked about side effects and underwent monthly blood checks. RESULTS: All patients were followed for at least 12 months. Mycophenolate mofetil proved to be effective with respect to the clinical course in 9 out of 10 eyes. All patients showed regression of inflammatory conjunctival alteration and improvement of their complaints. In one eye the inflammatory process restarted after surgery due to excessive symblephara had been performed. Gastrointestinal side effects were reported in the initial phase, e.g. lack of appetite, nausea and mild diarrhoea. CONCLUSIONS: Mycophenolate mofetil proved to be an effective immunosuppressant for the treatment of ocular cicatrical pemphigoid. Namely side effects were less severe and frequent compared to those known from other currently administered immunosuppressants. Longterm results and larger case numbers are needed to sustain these early results. PMID- 11392269 TI - [Quantitative analysis of tear protein profile for soft contact lenses--a clinical study]. AB - PURPOSE: Approximately 3% of the patients with refractive errors wear contact lenses. Recent studies revealed contradictory results regarding the changes of the tear film in contact lens wearers. The aim of this study was to analyze the influence of wearing contact lenses on the tear protein patterns. The study was performed using a new method to analyze quantitatively the tear proteins based on digital image analysis of electrophoretical separations followed by multivariate statistical calculations. METHODS: Two groups were examined: 121 healthy volunteers (KO) and 66 wearers of soft contact lenses (KL). The tear proteins were separated by sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS PAGE). For each electrophoretic lane, a densitograph was built by digital image analysis. Subsequently a multivariate analysis of discriminance was performed. RESULTS: The concentration of the tear proteins lactoferrin, sIgA, lysozyme and albumin was slightly reduced. In contrast, the concentration of lipocalin was increased in the tears of contact lens wearers (p < 0.07). However, including all peaks found in each electrophoretic lane, the analysis of discriminance found a statistical significant difference between KO and KL (Wilks' Lambda = 0.88; p < 0.0001). CONCLUSIONS: Wearing contact lenses leads to significant changes in the composition of tear-film proteins. However, the pathomechanism of this alteration of the tear film is still unclear. Because of the large number of contact lens wearers, these alterations should be examined further. PMID- 11392268 TI - [Ocular barostress and barotrauma. A study of 15 scuba divers]. AB - BACKGROUND: During SCUBA-diving, relative changes of the pressure in the diving mask, compared to the environmental pressure, are transmitted to the eye and to the periocular tissue. Barotrauma results from lack of pressure equilibration. MATERIAL AND METHODS: In 15 divers (6 experienced, 9 beginners) the pressure difference (delta-p) between inside and outside of the diving mask was measured every second during recreational SCUBA-diving. Data were transmitted wireless to a modified standard diving computer. An overall dive time of 323 minutes was analysed. RESULTS: Mean delta-p was 14.8 mbar (-44 to , std.dev. +/- 9), it was not dependent on the diving depth (r2 = 0.0004). Delta-p oscillated between 0 and 25 mbar (0-19 mm Hg), parallel to respiration. Negative delta-p values were 9.5 times more frequent in beginners than in experienced divers. Negative pressure peaks (changes > or = 10 mbar for more than 6 sec) occurred in the beginner group exclusively (p = 0.01). CONCLUSIONS: During SCUBA diving, ocular tissues undergo oscillating changes of environmental pressure, parallel to respiration. This may be important in eyes with previous surgery. Pressure peaks, leading to severe ocular barotrauma, are easily avoidable. PMID- 11392270 TI - [Supportive amblyopia treatment by means of computer games with background stimulation; a placebo controlled pilot study of 10 days]. AB - BACKGROUND: Computer programmes for visual stimulation may give new impulses to the field of amblyopia treatment by offering an option to shift the apparative visual training into the domestic sphere. Regarding this aspect we report on a placebo controlled study on a newly developed vision training consisting of a background stimulation by a drifting sinusoidal grating combined with a foreground game aimed to maintain the attention. MATERIALS AND METHODS: Fourteen amblyopia patients aged from 6 to 13 years participated in the study. Seven were allocated to a placebo and seven to a treatment group. Both groups had to train at the computer for a period of 10 working days by two sessions of about 20 minutes daily. Whilst the placebo group played in front of a neutral background, the treatment group did this with a drifting sinusoidal grating in the background. RESULTS: The treatment condition resulted in a greater increase of visual acuity than the placebo condition. Near vision improved in the treatment group from 0.20 (SD +/- 4.51 steps) to 0.39 (SD +/- 3.06 steps), i.e. by 3.0 steps of visual acuity (SD +/- 1.8 steps), in the placebo group from 0.14 (SD +/- 6.02 steps) to 0.17 (SD +/- 5.85 steps), i.e. by 0.8 steps of visual acuity (SD +/- 1.6 steps). Far vision improved in the treatment group from 0.29 (SD +/- 2.57 steps) to 0.44 (SD +/- 3.16 steps), i.e. by 1.9 steps of visual acuity (SD +/- 1.3 steps), in the placebo group from 0.24 (SD +/- 5.20 steps) to 0.28 (SD +/- 5.51 steps), i.e. by 0.7 steps of visual acuity (SD +/- 1.1 steps). CONCLUSIONS: Stimulation with drifting sinusoidal gratings improves the visual acuity of amblyopic eyes in a specific way. The effect might be accounted for by a synergy of spatial and temporal frequency in form vs. motion channels. A preliminary hypothesis is discussed and will be the subject of ongoing research. The presented method has been developed for the treatment of "delayed" amblyopia in the elder child. It is aimed to support and complement occlusion therapy. However, the reported results obtained over 10 days should be estimated only within the context of evaluation. By no means should the results be interpreted as a renewed pledge for a short-term treatment of amblyopia. PMID- 11392271 TI - [Human cultured lens cells. II. Characterization of established lens cell lines and test of their suitability for cytotoxicity tests]. AB - BACKGROUND: Human lens cells in culture may be valuable tools to discover cataractogenic risk factors. Here we report on the characterization of established human lens cells and their use in cytotoxicity tests. MATERIAL AND METHODS: Adhesion dependence was tested by an autoradiographic method. Soft agar test was performed to secure anchorage-independent growth. Laser scan microscopy was used for measuring the nucleus-cytoplasmic relationship. The cytoskeleton was visualized by fluorescence microscopy. Investigations on cytotoxicity were done by neutral red cytotoxicity assay and 3H-thymidine incorporation. The labeling index was determined by the BrdU-method. RESULTS: Two lens cell lines were transformed into fast growing cells. They are characterized by a partial loss of adhesion dependence, cell growth in soft agar, high cloning efficiency and reduced serum requirement. However, the nucleus-cytoplasmic relationship did not change very much in comparison to non-transformed lens cells. The organization of the cytoskeleton was cell shape dependent. The intermediate filaments were from the vimentin type. The established cell lines proved to be highly sensitive to ethanol and mitomycin C. CONCLUSIONS: The established human lens cell lines are well suited to screen for cytotoxic substances in vitro which might be cataractogenic risk factors in vivo. PMID- 11392272 TI - [Normal tension glaucoma, sleep apnea syndrome and nasal continuous positive airway pressure therapy--case report with a review of literature]. AB - BACKGROUND: In the pathogenesis of glaucoma, besides an elevated intraocular pressure (IOP), cardiovascular risk factors, such as arterial hypotension and hypertension, vasospasms, autoregulatory defects, atherosclerosis, and diabetes mellitus are of increasing importance, especially in normal tension glaucoma. Recently, there have been several reports of an additional risk factor: obstructive sleep apnea syndrome. METHODS: Literature review (Medline) and case report. RESULTS: The authors report on a 8 1/2 years follow-up of a 60-year-old patient with normal tension glaucoma. Despite successful pharmacological and surgical lowering of intraocular pressure a progressive glaucomatous damage with optic nerve atrophy and increasing visual field defects occurred. As a result of intensive investigations of possible cardiovascular risk factors, an obstructive sleep apnea syndrome was diagnosed. Since the beginning of therapy with nCPAP (nasal continuous positive airway pressure) more than 3 1/2 years ago, no further progression of glaucomatous optic nerve damage or visual field defects have been observed. CONCLUSIONS: In clinical practice, obstructive sleep apnea syndrome often is underdiagnosed. In patients suffering from glaucoma and obstructive sleep apnea syndrome, intraocular pressure lowering therapy may not be enough, whereas an additional nCPAP-therapy potentially could prevent the beginning/progression of glaucomatous optic nerve damage. PMID- 11392273 TI - [Conjunctival balloon cell nevi--light- and electronmicroscopic findings in 2 patients]. AB - CASE REPORTS: Two 19-year-old patients presented with small nasal conjunctival tumors. Histologic examination revealed subepithelial tumors consisting of large cells with deeply basophilic, small, centrally-placed nuclei and abundant, slightly eosinophilic cytoplasm with microvesicles. Electron microscopy showed premelanosomes in some of the balloon cells. CONCLUSION: The balloon cell nevus is a rare variant of the conjunctival nevi. The characteristic intracellular microvesicles are probably degenerative remnants of premelanosomes. PMID- 11392274 TI - [Persistence of bilateral pigmentary deposits in stromal cornea in a woman after a brief treatment with antidepressants]. AB - BACKGROUND: Neuropsychiatric agents may affect the eye in various ways. Phenothiazine commonly used in the treatment of depressive psychosis and schizophrenia may induce pigmentary deposits in the eyelids, conjunctiva, cornea, lens and retina. PATIENT: We report on a case of a 60-year-old female patient who presented with pigmentary changes in the cornea of unknown origin. We found bilateral, axial brown dustlike deposits in the posterior stroma of the cornea. Further ophthalmic investigation revealed no other abnormalities. Seven years ago the patient underwent therapy with fluphenazine, a phenothiazine derivative. The corneal changes were similar to those described for chlorpromazine, which is also a phenothiazine derivative. CONCLUSION: Phenothiazine may induce pigmentary changes of the eye. Ophthalmic followup in patients with phenothiazine therapy seems to be necessary. PMID- 11392275 TI - [Corneal ring opacity (Ascher ring)--a case report]. AB - PATIENT: A 63-year-old man without complaints presented with a bilateral symmetric opacity of the cornea between the mid and the outer periphery which was classified as Ascher's ring. CONCLUSIONS: Ascher's ring is a very rare entity. Characteristic slit lamp features allow its differentiation from other anular corneal opacities. Corneal or systemic alterations of the lipid metabolism can be discussed. Heredity can be suspected but could not be proved as yet. The visual prognosis is excellent. Therapy is not necessary. PMID- 11392276 TI - [Aversion of doctors to the subject of marketing]. PMID- 11392277 TI - [Physician's practice and trade. Part 2: Sharing of practice facilities]. PMID- 11392278 TI - [Your ultrasound diagnosis? Invagination of the small intestine]. PMID- 11392279 TI - [Ultrasound mammography]. AB - The importance of breast ultrasound in the determination of a palpable mass grew in the last years, to examine the radiologically dence breast or cysts it even became the first line method. The sonographic evaluation bases on the following criteria: internal echoes, shape of the tumor, posterior shadowing or enhancement, lateral shadows, compressibility and shifting. As there are only a few pathognomic and definite findings like anechoic cysts with smooth walls and sharp borders, the majority of palpable masses may be determined only approximately without final certainty. Therefore the diagnosis has to be evaluated by combining clinical and imaging analysis but final diagnosis remains for histological examination. PMID- 11392280 TI - [Value of ultrasound diagnosis in evaluating a round liver lesion]. AB - Ultrasonography is a modern noninvasive imaging technique with a high resolution, and high accuracy in diagnostic questions, highly accepted by patients, with low costs, and no side effects. There are only a few questions to witch the diagnostic ultrasound is unable to answer satisfactorily. New techniques complete the nowadays basic (B-Mode, M-Mode, Color and Power Doppler) ultrasonography, like intraoperative ultrasonography, quantitative ultrasonography or ultrasonography with contrast agents. Depending on the tumor species, the sensitivity and specificity in the diagnosis of a liver mass diagnostic is more than 95%. Also in difficult situations such as cirrhosis of the liver, the ultrasonography is equal to other imaging procedures. The basis of the success is, beside of a high quality ultrasonographic equipment, an excellent training, a skilled examinator owning a long experience as well as a standardized, reproducible examination technique with an excellent documentation of the findings. PMID- 11392281 TI - [Indications of fetal chromosome abnormalities in 1st trimester ultrasound]. AB - The measurement of the nuchal translucency and the evaluation of other sonographic signs in the first trimester scan allow a detection rate of 70-80% of aneuploid pregnancies, significantly more than with consideration of the maternal age alone (30%). The sonographic signs include the early growth retardation, deviations of the fetal heart rate, exomphalos, megacystis, holoprosencephaly and enlargement of the cisterna magna. Maternal serum biochemistry alone (PAPP-A and beta-hCG or alpha-fetoprotein, estriol and beta-hCG) detects about 65% of aneuploid pregnancies. The best individual risk estimation is based on maternal age, measurement of the nuchal translucency and the maternal biochemistry. PMID- 11392282 TI - [Ultrasound indications of fetal chromosome abnormalities in the 2nd trimester]. AB - The most common fetal chromosomal disorders have structural abnormalities, which can be detected during second trimester ultrasound examination. Major malformations, also known as hardmarkers, when single or in combination, should raise the suspicion for a specific syndrome. But it is known, that even more subtile findings can increase the background risk based on the maternal age, especially for Down syndrome. A combination of these so called "soft-markers" can be used for second trimester screening in a high and low risk population. Here were report on the sonographic features of the common chromosomal anomalies and their significance in prenatal diagnosis. PMID- 11392283 TI - [Ultrasound examination of the urogenital tract: indications and limits]. AB - Today, ultrasound examination has become an established standard in urological practice. Sonographic imaging has benefited immensely from the dramatic advancements in digital computer technology. We users now receive better, more accurate information and insights into the functional diagnosis of urological disorders. The ultrasound images are generated in real time and the patient is not exposed to any radiation. As far as costs are concerned, ultrasound instruments of good quality are available at affordable prices, even for general practitioners. This is one reason why ultrasound is being used more and more frequently, replacing more invasive, burdensome and expensive diagnostic techniques in multiple indications. The aim of this paper is to outline the growing number of indications for ultrasound examination in urology, while pointing out the specific limitations of this diagnostic method. PMID- 11392284 TI - [Transient osteoporosis in pregnancy. Transient osteoporosis of the right hip]. PMID- 11392285 TI - [Vitamin E is ineffective in treatment of late dyskinesias]. PMID- 11392286 TI - [Initial instability of different cages for fusion of the cervical spine]. AB - Purpose of the present investigation is to determine the biomechanical behaviour of different cages for monosegmental fusion of the cervical spine. Three commercially available cages (BAK, NOVUS, WING) representing the different principles of intercorporal implants and a combination of intercorporal bone graft together with anterior plating were tested for their resistance and sintering patterns under axial compression conditions. Therefore, FSU (functional spine-units) of 5-months old calfs were used. After preparation, the anterior fusion was performed by an orthopaedic surgeon. Specimen were mounted in a testing machine Zwick 1425 and axial load from 100 N up to 2000 N was applied. The compressed distance was measured and put into relation to the applied load. After that, the device was unloaded and the test was repeated another 2 times to determine the plastic deformation of implant and specimen. There was no significant difference to all of the constructs in the first compression. After repeated compression, the WING-cage, the NOVUS-cage and the plate-construct showed a constant compression pattern as expression of resting stable on the vertebral endplates, whereas the cylindrical BAK-cage had a decrease in compression distance, but increase in sintering into the vertebral body. Intercorporal implants that require the destruction of the vertebral endplates as described in the Cloward dowel-technique may have a higher risk of sintering into the vertebral body and therefore of developing progressive kyphosis. By attaching lateral supporting areas this risk can be reduced and the advantage of sponges contact for fusion is preserved. PMID- 11392287 TI - Non-invasive measurement of intracranial pressure changes by otoacoustic emissions (OAEs)--a report of preliminary data. AB - Up to now changes of intracranial pressure can only be objectively assessed by invasive measurement tools e.g. epidural transducers or intraventricular or intraparenchymatous catheters. Changes of intracranial pressure (ICP) are known to influence the inner ear since the subarachnoid space is linked to the perilymphatic space of the inner ear via the cochlear aquaeduct. A new method for assessing cochlear disorders is based on otoacoustic emissions (OAE) which are generated by the outer hair cells (OHCs) of the inner ear. The aim of the present study was to find out whether changes of intracranial pressure can be monitored by spontaneous otoacoustic emissions (SOAEs), transient evoked otoacoustic emissions (TEOAEs) and distortionproduct otoacoustic emissions (DPOAEs). SOAEs, TEOAEs and DPOAEs were measured in 12 young normally hearing subjects (volunteer group) in different body postures (horizontal, -30 degrees and +30 degrees supine position). In 5 patients undergoing continuous intraventricular pressure monitoring for the assessment of normal pressure hydrocephalus (NPH), DPOAEs were measured simultaneously in different body postures as well (patient group). At an increase of ICP the SOAE-level of the volunteer group decreased by -3.3 dB SPL (sound pressure level) and the TEOAE-level by -2.1 dB SPL. The DPOAEs showed a frequency dependent reduction of its level with maximal changes at the lowest frequency tested (f2 = 1 kHz; -7.9 dB SPL). In the patient group the ICP amounted to 19.2 cm H(2)0 and the DPOAE-level also decreased particularly at lower frequencies (-2.0 dB SPL). In conclusion otoacoustic emissions, particularly DPOAEs, may provide a new clinical tool for non-invasive monitoring of ICP. PMID- 11392288 TI - [Cerebral oxygen reactivity determination--a simple test with potential prognostic relevance]. AB - Brain tissue oximetry (ptiO2) using flexible micro-polarographic electrodes is a loco-regional approach to monitor oxygen supply to the injured brain, after neuronal damage. In patients after severe head injury (SHI), disturbances of CBF and CO2 related vasoconstriction have been demonstrated. CO2 reactivity testing may assist to determine outcome in these patients. Not much information is available on the preservation of vasoreactivity to arterial hyperoxia after neuronal damage. Therefore, we studied the response of ptiO2 in 7 piglets and in 14 patients on day one after trauma to 100% FiO2 ventilation (O2rea) and analyzed the 3 month outcome using the Glasgow-Outcome-Score (GOS). In the animal study, we placed a Paratrend 7 (P7) sensor for ptiO2 measurements in the non injured frontal white matter. The animals were anesthetized and mechanically ventilated. FiO2 was increased from 30 (+/- 5)% to 100% over a period of 5 minutes. In patients, we placed the P7 probe in the frontal lobe. FiO2 was increased from 35 (+/- 5)% to 100% over a period of 6 hours. O2rea was tested by calculating the percentage change of ptiO2 during 100% FiO2 ventilation, compared to the baseline value of 35% FiO2. By analyzing the patient outcome, we were able to define two patient populations according to the GOS at three month (Group I: favorable outcome [GOS 0-2]; Group II: poor outcome [GOS 3-4]). For the non-injured brain tissue in animals were revealed an O2rea = 0.21 (+/- 0.12). PATIENTS: Group I: O2rea = 0.4 (+/- 0.16); Group II: 0.9 (+/- 0.6). Group I and II were statistical significant different (p < 0.05; unpaired t-test). Oxygen reactivity in severely head patients is a simple test with prognostic value using ptiO2 measurement. These results may be explained by the close relationship of CBF disturbances to oxygen vasoreactivity after traumatic brain injury. The O2rea in animals without neuronal damage is smaller than in patients after SHI. We speculate, the animal data could be considered as normal value of O2rea in non injured brain tissue. PMID- 11392289 TI - [Neuro-navigation in the central area: impact on different surgical steps related to the location and various pathological processes]. AB - The neurosurgical treatment of space occupying processes in the central area bears a relatively high risk of either postoperative neurological deficits ("radical approach") or of residual tumor ("conservative approach"). Therefore, special techniques of intraoperative topographic orientation (image-guided surgery) play an important role here. The possible impact of neuronavigation on different neurosurgical steps (craniotomy, corticotomy, localization of the process, definition of borders of resection) was studied in relation to the site of pathology (extraaxial, intraaxial/superficial, intraaxial/deep) in 46 patients harbouring space occupying lesions of the central area. In intraaxial pathologies, additional electrophysiological monitoring was done. It could be shown, that in cases of deep seated processes, neuronavigation had the greatest impact on craniotomy, corticotomy and localization of the process, whereas the borders of resection were defined predominantly on the basis of differences in colour or consistency. In extraaxial pathologies, neuronavigation was of significance only for craniotomy; in intraaxial processes visible at the surface, it had an impact on craniotomy and--in a few cases--on definition of resection borders. In neurosurgery of intraaxial pathologies of the central area (particularly those not visible at the surface), the use of neuronavigation (or another method of intraoperative localization) in combination with neurophysiologic monitoring is strongly recommended. PMID- 11392290 TI - A cavernous hemangioma simulating an intracanalicular acoustic neurinoma--a case report. AB - A case of an intrameatal cavernous hemangioma is reported. The 53-year-old patient presented with decreased hearing and a slight 7th nerve palsy on the left. Clinical features and preoperative radiological appearances were indistinguishable from those of an acoustic neurinoma. Though, DD the involvement of all three nerves (N. facialis, N. cochlearis, N. vestibularis) should have lead to another entity. The tumour showed intraoperatively no relationship to the 8th cranial nerve, but was very adherent to the facial nerve. The macroscopic appearance differed to the usual aspect of an acoustic neurinoma. The final diagnosis was made after the operation with histopathological methods. The clinical features and pathology of this type of tumour are discussed. PMID- 11392291 TI - Artifacts in musculoskeletal magnetic resonance imaging: identification and correction. AB - A large number of artifacts occur in magnetic resonance (MR) imaging of the musculoskeletal system. These artifacts may potentially affect the quality of MR images, and may also simulate pathologic conditions and produce pitfalls in interpretation. Motion artifacts may be periodic or random. Protocol-error artifacts include saturation, wraparound, radiofrequency (RF) interference, shading, and partial volume averaging artifacts. Truncation artifacts occur when the number of phase-encoding steps of high spatial frequencies is insufficient (or under-sampled) for faithful reproduction of the true anatomic detail of the original image. Chemical shift artifacts are due to the protons in fat being mismapped relative to water protons. Susceptibility artifacts occur at the interfaces of structures with different magnetic susceptibilities. Artifacts special to the musculoskeletal system include the magnic angle phenomenon and spurious signal induced at very short echo times, both of which affect anisotropic structures such as tendon, ligament, and cartilage. Recognition and, if possible, correction of these artifacts are an important aspect of practical musculoskeletal MR imaging. PMID- 11392292 TI - Florid reactive periostitis and bizarre parosteal osteochondromatous proliferation: pre-biopsy imaging evolution, treatment and outcome. AB - OBJECTIVE: To report on the imaging evolution of florid reactive periostitis (FRP) and bizarre parosteal osteochondromatous proliferation (BPOP) of the phalanges of the hands from prospective diagnosis to operation and on postsurgical outcome. DESIGN AND PATIENTS: Three patients (2 female, 1 male; age range 11-34 years) presented with a swollen digit of the hand. Following presumptive radiographic diagnosis of FRP, they were closely observed both clinically and radiographically until operation. All three patients had radiographs of the involved digit, and one patient had an MR imaging examination. The interval between presumptive diagnosis and operation ranged from 2 to 8 months. Following operation, the patients have been clinically followed for 9-13 months (mean 10 months). RESULTS: In each of the patients, maturing of periosteal reaction without bone destruction was observed within 1-2 weeks of the presumptive diagnosis of FRP. Periosteal reaction was initially minimal in relation to the extent of soft tissue swelling and subsequently became more florid. In one patient, the lesion ossified, became adherent to the phalanx, and had an "osteochondromatous" appearance. In another patient, periosteal reaction was seen on both sides of the phalanx with an intact phalanx. In the sole patient who had MR imaging, edema was seen in the phalanx distal to the symptomatic site and the metacarpal proximal to the symptomatic site. CONCLUSIONS: Close clinical and radiographic correlation permits an accurate pre-biopsy diagnosis of FRP. The first follow-up radiograph taken within 2 weeks usually provides re-assurance of the accuracy of the diagnosis. FRP may progress to BPOP. Arbitrary antibiotic treatment can be avoided, and a planned surgical approach can be adopted. PMID- 11392293 TI - Assessment of instability of the long head of the biceps tendon by MRI. AB - OBJECTIVE: To determine whether MRI can identify instability of the long head of the biceps tendon (LBT) in the rotator interval. DESIGN AND PATIENTS: A retrospective review was carried out of 19 patients, all arthroscopically examined, nine of whom had surgically confirmed instability of the LBT. RESULTS: A LBT perched on the lesser tuberosity correctly indicated all nine cases of instability with one false positive. In six of seven cases where the LBT was oval in shape, no instability of the biceps tendon existed, whereas LBT instability was present in eight of 12 patients with a flat long head of the biceps tendon. In seven of eight acutely angled intertubercular sulci there was no instability of the LBT while eight of 11 obtusely angled sulci were associated with LBT instability. By consensus impression, instability of the LBT could be determined with 67% sensitivity, 90% specificity, 86% positive predictive value, and 75% negative predictive value. CONCLUSIONS: A flat LBT perched on the lesser tuberosity with an obtusely angled intertubercular sulcus suggests the diagnosis of instability of the LBT in the correct clinical setting. PMID- 11392294 TI - Radiological features of 24 periosteal chondrosarcomas. AB - OBJECTIVE: To report the imaging findings of 24 periosteal chondrosarcomas diagnosed, staged, treated and followed in a single institution, to analyze and define their pattern, and discuss their practical consequences. DESIGN AND PATIENTS: Plain films, 16 CT examinations and four MRI examinations were reviewed, and compared with the histological evaluation. RESULTS: There were 20 men and four women, aged from 17 to 65 years. Twelve lesions involved the distal femoral metaphyses (8 posteriorly), five the proximal humerus, two the proximal metaphyses of the femur and two of the tibia, two the humeral shafts and one the iliac wing. Size varied from 4 to 11 cm. The cortex was always involved (thick, 15; thin, 13). Typical cartilaginous calcifications and cartilaginous lobules were very frequent. Radial thick periosteal bone formations (n = 6) indicated calcifications between the lobules of cartilage. Medullary involvement was rare (n = 2). All patients are alive and free of disease. CONCLUSIONS: Recognizing periosteal chondrosarcoma is of paramount importance because the prognosis is excellent after adequate local surgery alone. The patterns of other surface tumors of bone are usually different. PMID- 11392295 TI - MRI of spinal hardware: comparison of conventional T1-weighted sequence with a new metal artifact reduction sequence. AB - OBJECTIVE: This study was designed to compare diagnostic quality of MR images of patients with spinal hardware acquired using a conventional T1-weighted spin-echo sequence and a new metal artifact reduction sequence (MARS). CONCLUSION: The new MARS sequence effectively reduces the degree of tissue-obscuring artifact produced by spinal fixation hardware and subjectively improves image quality compared with the conventional T1-weighted spin-echo sequence. PMID- 11392296 TI - Treatment of epithelioid hemangioendothelioma of bone using a novel combined approach. AB - Radiofrequency (RF) treatment has been used for a variety of malignant and benign conditions. However, treatment of a bone malignancy has yet to be reported. The authors present a 21-year old woman with multifocal epithelioid hemangioendothelioma (EH) treated by a combination of surgical excision, chemotherapy and four RF ablations. Follow-up radiographs of the RF-treated sites reveal no evidence of recurrent disease 71, 58, 49 and 33 months, respectively, after treatment. PMID- 11392297 TI - Blood-pool scintigraphic diagnosis of fractured lumbar vertebral hemangioma. AB - A 57-year-old woman complained of lumbago of 1 year's duration. Radiographs showed a compression fracture of the third lumbar vertebra. CT and MR images revealed an enhancing mass confined to the vertebral body suggestive of a malignant process. A blood-pool scintigram with 99mTc-human serum albumin combined with DTPA (HSA-D) revealed marked accumulation. This strongly suggested a hemangioma, which was confirmed by biopsy. PMID- 11392298 TI - Epithelioid hemangioma of bone. AB - Although no specific radiographic appearance has emerged to date for the epithelioid subtype of hemangioma, these lesions most typically exhibit well defined osteolysis. Other relatively common features include surrounding sclerosis, cortical expansion and cortical destruction. We present a case of epithelioid hemangioma of the spine with an unusual radiological appearance which to our knowledge has not previously been reported: diffuse sclerosis of the involved vertebral body. The diffuse sclerosis seen in this case resembles the osteosclerotic process seen not only in benign entities such as subacute and chronic osteomyelitis, but also in malignant lesions such as osteoblastic metastatic disease and lymphoma. PMID- 11392299 TI - Intra-articular lipoma with osteochondroid metaplasia in the knee joint. AB - We describe a case of lipoma with osteochondroid metaplasia in the knee joint. Although the location of the lesion and radiographic findings were unusual, computed tomography and magnetic resonance imaging were useful in characterizing adipose, cartilaginous and osseous tissue components within the lesion. PMID- 11392300 TI - Anterior lumbar interbody fusion. AB - A new technique for lumbar fusion surgery incorporates threaded interbody cages. In contrast to its posterior counterpart, the anterior approach to this surgery appears to result in better fusion, less morbidity and quicker operations. This article reviews spinal anatomy and fusion and discusses the indications for anterior lumbar interbody fusion. The radiographer's role in diagnostic imaging, surgery and post-operative routines is chronicled. Radiation dose considerations are addressed and suggestions for improving follow-up assessment are offered. PMID- 11392301 TI - Stereotactic breast biopsy. AB - This overview of stereotactic breast biopsy describes the procedure's advantages over surgical biopsy as well as contraindications for its use. Patient and procedural considerations are discussed, along with the equipment used, technologist qualifications and imaging and positioning techniques. The article concludes with a discussion of quality assurance requirements. PMID- 11392302 TI - Radiography of bone tumors and lesions. AB - This Directed Reading reviews the biology and classification of bone tumors, forms of primary bone tumors and techniques for imaging these lesions. PMID- 11392303 TI - MS: the body at war with itself. PMID- 11392304 TI - Are managers wasting words on praise? AB - Praise can be a powerful form of affirmation and a motivator. However, praise also can be given in a way that negates its benefits. To avoid wasting words and maximize the benefits of praise, it must be offered in the right manner and at the right time. PMID- 11392305 TI - Seeing the future in three dimensions. PMID- 11392306 TI - Online teaching: design and techniques. PMID- 11392307 TI - Lateral lumbar ghost. PMID- 11392308 TI - Outcomes research: who needs it? PMID- 11392309 TI - [Recommendations for hospital units and instrumentation in pediatric anesthesia]. PMID- 11392310 TI - [A score board for anesthesia services]. PMID- 11392311 TI - [Postoperative surveillance units: new specifications]. PMID- 11392312 TI - A case of de novo early erythroblastic leukemia supporting a proposal of AML M6 'variant'. PMID- 11392313 TI - Reversible portal vein thrombosis complicating induction therapy of acute promyelocytic leukemia. PMID- 11392314 TI - Platelet function and total length of intracoronary stents: is there a correlation? PMID- 11392315 TI - Central lamellar keratitis. PMID- 11392316 TI - Healing of closed femoral shaft fractures treated with the AO unreamed femoral nail. A comparative study with the AO unreamed femoral nail. (Injury, 2000; 31:367-371). PMID- 11392317 TI - Clinical photography in patient referral. PMID- 11392318 TI - Peru moves to stop multidrug-resistant tuberculosis. PMID- 11392319 TI - Report on the first European Doctoral Conference in Nursing Science in Berlin. PMID- 11392320 TI - May 2000 supplement on preventing occupational injuries. PMID- 11392321 TI - From my perspective...the meaning of professionalism. PMID- 11392322 TI - Further characterization of the autism susceptibility locus AUTS1 on chromosome 7q. AB - Autism is a neurodevelopmental disorder that usually arises on the basis of a complex genetic predisposition. The most significant susceptibility region in the first whole genome screen of multiplex families was on chromosome 7q, although this linkage was evident only in UK IMGSAC families. Subsequently all other genome screens of non-UK families have found some evidence of increased allele sharing in an overlapping 40 cM region of 7q. To further characterize this susceptibility locus, linkage analysis has now been completed on 170 multiplex IMGSAC families. Using a 5 cM marker grid, analysis of 125 sib pairs meeting stringent inclusion criteria resulted in a multipoint maximum LOD score (MLS) of 2.15 at D7S477, whereas analysis of all 153 sib pairs generated an MLS of 3.37. The 71 non-UK sib pairs now contribute to this linkage. Linkage disequilibrium mapping identified two regions of association-one lying under the peak of linkage, the other some 27 cM distal. These results are supported in part by findings in independent German and American singleton families. PMID- 11392324 TI - Parentage and heritage of dendritic cells. PMID- 11392325 TI - A novel BCR-ABL fusion transcript (e15a2) in 2 patients with atypical chronic myeloproliferative syndrome. PMID- 11392326 TI - Long-term follow-up of a randomized trial comparing the combination of cyclophosphamide with total body irradiation or busulfan as conditioning regimen for patients receiving HLA-identical marrow grafts for acute myeloblastic leukemia in first complete remission. PMID- 11392327 TI - Intensive chemotherapy versus bone marrow transplantation in pediatric acute myeloid leukemia: a matter of controversies. PMID- 11392328 TI - Intensive chemotherapy and bone marrow transplantation for children with acute myeloid leukemia. PMID- 11392329 TI - Treatment of children with acute myeloid leukemia. PMID- 11392330 TI - Screening for c-mpl mutations in patients with congenital amegakaryocytic thrombocytopenia identifies a polymorphism. PMID- 11392331 TI - Frequent detection of rising cytomegalovirus antigenemia after allogeneic stem cell transplantation following a regimen containing antithymocyte globulin. PMID- 11392332 TI - Adolescent neuropsychological development after early right prefrontal cortex damage. AB - Adolescent development and pattern of recovery are described for a 15-year-old boy who sustained extensive right dorsolateral prefrontal cortex damage at age 7 from rupture and surgical treatment of a deep arteriovenous malformation. Follow up evaluations at 4 years and most recently 8 years after illness have shown clear improvement in social-behavioral and almost all cognitive areas initially assessed. He demonstrated resolution of left hemispatial neglect and other visuospatial impairments in working memory, design fluency, and planning and organization. However, at the 8-year follow-up interval, an acquired form of attention deficit disorder remains evident. We hypothesized that this is the likely cause of comparatively lower scores in general intelligence, verbal learning and memory, discourse, and processing speed, that at the 4-year follow up interval. New measures of emotional face and voice recognition showed only minor difficulties, principally in identifying vocal disgust and fear. Social and psychological maturation has continued to improve, with no evidence of developmental arrest or pervasive social impairment, although the individual is confused at times by complexities and nuances of social interaction. The pattern of findings 8 years after early right dorsolateral prefrontal cortex damage suggests remarkable recovery of primary visuospatial and social impairments, but lingering and somewhat worsening performance deficits which may be due to attentional difficulties and impulsive responding. Treatment of the attentional difficulties is currently being investigated. PMID- 11392333 TI - [Developments in dentistry in the twentieth century 5. Mouth diseases and oral surgery]. PMID- 11392334 TI - Evidence-based medicine. PMID- 11392335 TI - Explosive outbursts associated with methylphenidate. PMID- 11392336 TI - The juvenile-as-adult-criminal debate. PMID- 11392337 TI - Adderall, the atypicals, and weight gain. PMID- 11392338 TI - Administrative prevalence of autism in the Texas school system. PMID- 11392339 TI - Pemoline hepatotoxicity and postmarketing surveillance. AB - OBJECTIVE: To review the numerous reports of hepatotoxic adverse drug reactions (ADRs) ascribed to pemoline that were sent to the U.S. Food and Drug Administration (FDA) between 1975 and 1996 and to describe the medical community's lack of awareness of these reports. METHOD: All ADR reports from 1975 through 1996 wherein pemoline was the suspect agent were obtained from the FDA MedWatch Internet site, and some details of nine pemoline-related deaths in youths were obtained directly from the FDA. The published literature on this subject was fully reviewed. RESULTS: (1) In premarketing clinical trials with pemoline in the early 1970s, hepatic abnormalities were noted in enzyme levels (1%-3% of youths receiving maintenance treatment), during rechallenges (6 of 6), and in biopsies (2 of 2). (2) Between 1975 and 1989, 12 cases of jaundice and 6 deaths in youths ascribed to pemoline hepatotoxicity were reported to the FDA. (3) The first medical literature report of a serious ADR ascribed to pemoline was in a 1989 letter to the editor. (4) Physicians generally only became aware of serious pemoline hepatotoxicity in December 1996. (5) Pemoline use increased until 1997. CONCLUSION: Limitations in postmarketing surveillance and public reporting in the United States, particularly in the 1980s, largely accounted for delays in an appropriate response to pemoline hepatotoxicity. PMID- 11392340 TI - Behavioral and neural consequences of prenatal exposure to nicotine. AB - OBJECTIVE: To review evidence for the neurodevelopmental effects of in utero exposure to nicotine. Concerns about long-term cognitive and behavioral effects of prenatal exposure to nicotine arise from reports of increased rates of disruptive behavioral disorders in children whose mothers smoked during pregnancy. The relatively high rate of tobacco smoking among pregnant women (25% of all pregnancies in the U.S.) underlines the seriousness of these concerns. METHOD: This review examines the largest and most recent epidemiological and clinical studies that investigated the association of prenatal nicotine exposure with health, behavioral, and cognitive problems. Because of the numerous potential confounding variables in human research, findings from animal studies, in which environmental factors are strictly controlled, are also discussed. Finally, neural and molecular mechanisms that are likely to underlie neurodevelopmental disruptions produced by prenatal nicotine exposure are outlined. RESULTS: A dose-response relationship between maternal smoking rates and low birth weight (potentially associated with lower cognitive ability) and spontaneous abortion is consistently found, whereas long-term developmental and behavioral effects in the offspring are still controversial, perhaps because of the difficulty of separating them from other genetic and environmental factors. Despite the wide variability of experimental paradigms used in animal studies, common physical and behavioral effects of prenatal exposure to nicotine have been observed, including low birth weight, enhanced locomotor activity, and cognitive impairment. Finally, disturbances in neuronal pathfinding, abnormalities in cell proliferation and differentiation, and disruptions in the development of the cholinergic and catecholaminergic systems all have been reported in molecular animal studies of in utero exposure to nicotine. CONCLUSIONS: Prenatal exposure to nicotine may lead to dysregulation in neurodevelopment and can indicate higher risk for psychiatric problems, including substance abuse. Knowledge of prenatal exposure to nicotine should prompt child psychiatrists to closely monitor at-risk patients. PMID- 11392341 TI - Understanding AIDS-risk behavior among adolescents in psychiatric care: links to psychopathology and peer relationships. AB - OBJECTIVE: Severely mentally ill youths are at elevated risk for human immunodeficiency virus infection, but little is known about acquired immunodeficiency syndrome (AIDS) risk behavior in adolescents who seek outpatient mental health services or about the links between psychiatric problems and particular high-risk behaviors. This pilot study used structural equation modeling to conduct a path analysis to explore the direct and indirect effects of adolescent psychopathology on risky sex, drug/alcohol use, and needle use. METHOD: Ethnically diverse youths (N = 86) and their caregivers who sought outpatient psychiatric services in Chicago completed questionnaires of adolescent psychopathology. Youths reported their relationship attitudes, peer influence, sexual behavior, and drug/alcohol use. RESULTS: Different AIDS-risk behaviors were associated with distinct forms of adolescent psychopathology (e.g., delinquency was linked to drug/alcohol use, whereas aggression was related to risky sexual behavior), and peer influence mediated these linkages. Some patterns were similar for caregiver- and adolescent-reported problems (e.g., peer influence mediated the relation between delinquency and drug/alcohol use), but others were different (e.g., caregiver-reported delinquency was associated with risky sex, whereas adolescent-reported delinquency was not). CONCLUSIONS: Findings underscore the complexity of factors (types of informants and dimensions of psychopathology) that underlie AIDS risk in troubled youths, and they offer specific directions for designing and implementing uniquely tailored AIDS prevention programs, for example, by targeting delinquent behavior and including high-risk peers and important family members in interventions. PMID- 11392342 TI - Case series: amantadine open-label treatment of impulsive and aggressive behavior in hospitalized children with developmental disabilities. AB - OBJECTIVE: A growing body of literature implicates interactions between glutamatergic and neostriatal dopaminergic neurotransmitter systems in the development and expression of impulsivity, hyperactivity, and stereotypy. Amantadine hydrochloride, a drug used in young children for influenza prophylaxis, acts both as an indirect dopamine agonist as well as an N-methyl-D aspartate (NMDA) receptor antagonist. Thus an open clinical trial of this medication for the treatment of symptoms of impulse control disorders in children was performed. METHOD: A total of eight children (seven with neurodevelopmental disorders and all inpatients) with target behaviors refractory to other treatments were selected after parental informed consent. All patients were male and ranged in age from 4 to 12 years. Outcome was based on subjective consensus clinical ratings by the multidisciplinary treatment team. RESULTS: For four of the children, amantadine was associated with marked clinical improvement. In the remainder, improvement was also observed. Amantadine was well tolerated. CONCLUSIONS: On the basis of this experience, it appears that amantadine hydrochloride or related NMDA antagonists may warrant additional study in this and related populations. PMID- 11392344 TI - Posterior fossa magnetic resonance imaging in autism. AB - OBJECTIVE: To determine whether the sizes and volumes of the posterior fossa structures are abnormal in non-mentally retarded autistic adolescents and adults. METHOD: Volume measurements of the cerebellum, vermis, and brainstem were obtained from coronal magnetic resonance imaging scans in 16 autistic subjects and 19 group-matched healthy controls. For the purpose of comparison with previous studies, area measurements of the midbrain, pons, medulla, total cerebellar vermis, and its three subregions were also obtained from a larger sample of 22 autistic males (mean age: 22.4 years; range: 12.2-51.8 years) and 22 individually matched controls (mean age 22.4 years; range: 12.9-52.2 years). RESULTS: The total volume of the cerebellum and the cerebellar hemispheres were significantly larger in the autistic subjects with and without correcting for total brain volume. Volumes of the vermis and the brainstem and all area measurements did not differ significantly between groups. CONCLUSIONS: There is an increase in the volume of the cerebellum in people with autism consistent with the increase in regional and total brain size reported in this developmental disorder. This finding is also concordant with evidence of cerebellar abnormalities from neuropathological and neuropsychological studies that point to the role of this structure, as part of a complex neural system, in the pathophysiology of autism. PMID- 11392343 TI - Double-blind, placebo-controlled study of amantadine hydrochloride in the treatment of children with autistic disorder. AB - OBJECTIVE: To test the hypothesis that amantadine hydrochloride is a safe and effective treatment for behavioral disturbances--for example, hyperactivity and irritability--in children with autism. METHOD: Thirty-nine subjects (intent to treat; 5-19 years old; IQ > 35) had autism diagnosed according to DSM-IV and ICD 10 criteria using the Autism Diagnostic Interview-Revised and the Autism Diagnostic Observation Schedule-Generic. The Aberrant Behavior Checklist Community Version (ABC-CV) and Clinical Global Impressions (CGI) scale were used as outcome variables. After a 1-week, single-blind placebo run-in, patients received a single daily dose of amantadine (2.5 mg/kg per day) or placebo for the next week, and then bid dosing (5.0 mg/kg per day) for the subsequent 3 weeks. RESULTS: When assessed on the basis of parent-rated ABC-CV ratings of irritability and hyperactivity, the mean placebo response rate was 37% versus amantadine at 47% (not significant). However, in the amantadine-treated group there were statistically significant improvements in absolute changes in clinician-rated ABC-CVs for hyperactivity (amantadine -6.4 versus placebo -2.1; p = .046) and inappropriate speech (-1.9 versus 0.4; p = .008). CGI scale ratings were higher in the amantadine group: 53% improved versus 25% (p = .076). Amantadine was well tolerated. CONCLUSIONS: Parents did not report statistically significant behavioral change with amantadine. However, clinician-rated improvements in behavioral ratings following treatment with amantadine suggest that further studies with this or other drugs acting on the glutamatergic system are warranted. The design of these and similar drug trials in children with autistic disorder must take into account the possibility of a large placebo response. PMID- 11392345 TI - Adult psychosocial outcome of prepubertal major depressive disorder. AB - OBJECTIVE: To compare adult psychosocial functioning (PSF) of subjects with prepubertal major depressive disorder (PMDD) to a normal comparison (NC) group. METHOD: PSF of subjects with PMDD (n = 72) and of NC subjects (n = 28) was compared after prospective follow-up to adulthood. These 100 subjects were 90.9% of the baseline 110 subjects who participated in the "Nortriptyline in Childhood Depression: Follow-up Study." Research nurses who were blind to group status conducted telephone interviews using the Longitudinal Interval Follow-up Evaluation (LIFE) to obtain PSF data. RESULTS: At follow-up, the PMDD group was 20.7+/-2.0 and the NC subjects were 20.9+/-2.2 years old. The PMDD subjects were 10.3+/-1.5 years old at baseline. Time between baseline and follow-up was 9.9+/ 1.5 years. In the PMDD group, subjects with MDD, bipolar disorder, or substance use disorders during the previous 5 years had significantly worse PSF than NC subjects. These PSF impairments included significantly worse relationships with parents, siblings, and friends; significantly worse functioning in household, school, and work settings; and worse overall quality of life and global social adjustment. CONCLUSIONS: Although combined treatments for PMDD have little scientific basis, multimodality regimens seem prudent until definitive treatment data become available. PMID- 11392346 TI - Predicting girls' conduct disorder in adolescence from childhood trajectories of disruptive behaviors. AB - OBJECTIVE: To examine girls' developmental trajectories of disruptive behaviors during the elementary school years and to predict conduct disorder symptoms and diagnosis in adolescence with trajectories of these behaviors. METHOD: The sample was 820 girls from the province of Quebec followed over 10 years (1986-1996). A semiparametric mixture model was used to describe girls' developmental trajectories of teacher-rated disruptive behaviors between the ages of 6 and 12 years. The trajectories were used to predict conduct disorder symptoms and diagnosis when the girls were on average 15.7 years. RESULTS: Four groups of girls following trajectories with distinct levels of disruptive behaviors were identified: a low, medium, medium-high, and high trajectory. Prediction with the trajectories indicated that girls on the medium, medium-high, and high trajectories reported a significantly higher number of conduct disorder symptoms in adolescence. However, only the girls on the medium-high and high trajectories were at significantly higher risk to meet DSM-III-R criteria for conduct disorder, compared with girls in the low group (odds ratio: 4.46). More than two thirds of the girls with conduct disorder were in the medium or higher-level trajectories. CONCLUSION: The results suggest that there is an early-onset type of conduct disorder in girls. PMID- 11392347 TI - Prospective, longitudinal study of tic, obsessive-compulsive, and attention deficit/hyperactivity disorders in an epidemiological sample. AB - OBJECTIVE: Understanding the interrelatedness of tics, obsessive-compulsive disorder (OCD), and attention-deficit/hyperactivity disorder (ADHD) has been complicated by studying only cross-sectional samples of clinically referred subjects. The authors report the cross-sectional and longitudinal associations of these disorders in an epidemiological sample of children followed prospectively into early adulthood. METHOD: Structured diagnostic interview information was acquired on 976 children, aged 1 to 10 years, who were randomly selected from families living in upstate New York in 1975. Reassessments were acquired in 776 of these subjects 8, 10, and 15 years later. Diagnostic prevalences were estimated at each time point. The associations among tics, OCD, and ADHD were assessed within and across time points, as were their associations with comorbid illnesses and demographic risk factors. RESULTS: In temporal cross-section, tics and ADHD symptoms were associated with OCD symptoms in late adolescence and early adulthood after demographic features and comorbid psychiatric symptoms were controlled. In prospective analyses, tics in childhood and early adolescence predicted an increase in OCD symptoms in late adolescence and early adulthood. ADHD symptoms in adolescence predicted more OCD symptoms in early adulthood, and OCD in adolescence predicted more ADHD symptoms in adulthood. The associations of tics with ADHD were unimpressive in temporal cross-section and were not significant in prospective analyses. Tics, OCD, and ADHD shared numerous complex associations with demographic and psychopathological risk factors. ADHD was associated with lower IQ and lower social status, whereas OCD was associated with higher IQ. CONCLUSIONS: Tics and OCD were significantly associated in this sample, as were OCD and ADHD. These findings are in general consistent with those from family studies, and they help to define the natural history, comorbid illnesses, and interrelatedness of these conditions. PMID- 11392348 TI - Aggression and psychiatric comorbidity in children with hypothalamic hamartomas and their unaffected siblings. AB - OBJECTIVE: To assess aggression and psychiatric comorbidity in a sample of children with hypothalamic hamartomas and gelastic seizures and to assess psychiatric diagnoses in siblings of study subjects. METHOD: Children with a clinical history of gelastic seizures and hypothalamic hamartomas (n = 12; age range 3-14 years) had diagnoses confirmed by video-EEG and head magnetic resonance imaging. Structured interviews were administered, including the Diagnostic Interview for Children and Adolescents-Revised Parent Form (DICA-R-P), the Test of Broad Cognitive Abilities, and the Vitiello Aggression Scale. Parents were interviewed with the DICA-R-P about each subject and a sibling closest in age without seizures and hypothalamic hamartomas. Patients were seen from 1998 to 2000. RESULTS: Children with gelastic seizures and hypothalamic hamartomas displayed a statistically significant increase in comorbid psychiatric conditions, including oppositional defiant disorder (83.3%) and attention deficit/hyperactivity disorder (75%). They also exhibited high rates of conduct disorder (33.3%), speech retardation/learning impairment (33.3%), and anxiety and mood disorders (16.7%). Significant rates of aggression were noted, with 58% of the seizure patients meeting criteria for the affective subtype of aggression and 30.5% having the predatory aggression subtype. Affective aggression was significantly more common (p < .05). Unaffected siblings demonstrated low rates of psychiatric pathology on semistructured parental interview and no aggression as measured by the Vitiello Aggression Scale. CONCLUSIONS: Children with hypothalamic hamartomas and gelastic seizures had high rates of psychiatric comorbidity and aggression. Parents reported that healthy siblings had very low rates of psychiatric pathology and aggression. PMID- 11392349 TI - Social impairment in girls with ADHD: patterns, gender comparisons, and correlates. AB - OBJECTIVE: To investigate social impairment in girls with attention deficit/hyperactivity disorder (ADHD), compare the social functioning of boys and girls with ADHD, and explore the association between social dysfunction and conditions comorbid with ADHD. METHOD: Four groups of index children were studied: 267 children (127 girls) with ADHD and 234 non-ADHD comparison children (114 girls). Groups were compared on social functioning, psychopathology, and demographic characteristics. RESULTS: Girls with ADHD manifested significant deficits in interpersonal functioning compared with girls without ADHD and evidenced a similar degree of social impairment compared with boys with ADHD. ADHD and associated comorbid disorders were significant correlates of specific domains of social dysfunction in boys and girls with ADHD. CONCLUSIONS: Interpersonal deficits are a major correlate of ADHD, irrespective of gender, and appear to stem from the behaviors associated with ADHD as well as behaviors characteristic of conditions comorbid with ADHD. PMID- 11392350 TI - Factor and latent class analysis of DSM-IVADHD symptoms in a school sample of Brazilian adolescents. AB - OBJECTIVE: To evaluate the validity of the multidimensional construct proposed by DSM-IV for the diagnosis of attention-deficit/hyperactivity disorder (ADHD) in a school sample of young Brazilian adolescents. METHOD: An instrument including all 18 DSM-IVADHD symptoms was administered to 1,013 students aged 12 to 14 years at 64 state schools by trained research assistants. Each symptom was rated on a Likert scale with five levels of severity (never, almost never, sometimes, frequently, and always). RESULTS: Using an exploratory factor analytic approach (principal components analysis), two factors were extracted. Factor I (hyperactivity-impulsivity) comprised eight DSM-IV hyperactive-impulsive symptoms with loadings > or =0.40. Factor II (inattention) included also eight DSM-IV symptoms of inattention. The two factors explained 34% of the total variance and had an interfactor correlation of 0.45. Latent class analysis demonstrated similar classes in males and females, but class structures were markedly different from previous analyses of parent report data. CONCLUSION: The findings support the appropriateness of the multidimensional construct introduced by DSM IV in the diagnosis of ADHD in a different culture but emphasize the possible impact of different reporters on the results of structural model-testing. PMID- 11392351 TI - Offender and offense characteristics of a nonrandom sample of adolescent mass murderers. AB - OBJECTIVE: The authors conducted a descriptive, archival study of adolescent (< or =19 years of age) mass murderers-subjects who intentionally killed three or more victims in one event-to identify demographic, clinical, and forensic characteristics. METHOD: A nonrandom sample of convenience of adolescent mass murderers was utilized. RESULTS: Thirty-four subjects, acting alone or in pairs, committed 27 mass murders between 1958 and 1999. The sample consisted of males with a median age of 17. A majority were described as "loners" and abused alcohol or drugs; almost half were bullied by others, preoccupied with violent fantasy, and violent by history. Although 23% had a documented psychiatric history, only 6% were judged to have been psychotic at the time of the mass murder. Depressive symptoms and historical antisocial behaviors were predominant. There was a precipitating event in most cases--usually a perceived failure in love or school- and most subjects made threatening statements regarding the mass murder to third parties. The majority of the sample clustered into three types: the family annihilator, the classroom avenger, and the criminal opportunist. CONCLUSIONS: The adolescent mass murderer is often predatorily rather than affectively violent and typically does not show any sudden or highly emotional warning signs. Although the act of mass murder is virtually impossible to predict because of its extremely low frequency, certain clinical and forensic findings can alert the clinician to the need for further, intensified primary care, including family, school, community, law enforcement, and mental health intervention. PMID- 11392352 TI - Measurement validity. PMID- 11392353 TI - Genetics of childhood disorders: XXVII. Genes and cognition in Williams syndrome. PMID- 11392355 TI - Re: characterization of the ventricular conduction system in the developing mouse heart. PMID- 11392356 TI - The structures of Langmuir-Blodgett films of fatty acids and their salts. AB - Recent advances in several experimental techniques have enabled detailed structural information to be obtained for floating (Langmuir) monolayers and Langmuir-Blodgett films. These techniques are described briefly and their application to the study of films of fatty acids and their salts is discussed. Floating monolayers on aqueous subphases have been shown to possess a complex polymorphism with phases whose structures may be compared to those of smectic mesophases. However, only those phases that exist at high surface pressures are normally used in Langmuir-Blodgett (LB) deposition. In single LB monolayers of fatty acids and fatty acid salts the acyl chains are in the all-trans conformation with their long axes normal to the substrate. The in-plane molecular packing is hexagonal with long-range bond orientational order and short-range positional order: known as the hexatic-B structure. This structure is found irrespective of the phase of the parent floating monolayer. The structures of multilayer LB films are similar to the structures of their bulk crystals, consisting of stacked bilayer lamellae. Each lamella is formed from two monolayers of fatty acid molecules or ions arranged head to head and held together by hydrogen bonding between pairs of acids or ionic bonding through the divalent cations. With acids the acyl chains are tilted with respect to the substrate normal and have a monoclinic structure, whereas the salts with divalent cations may have the chains normal to the substrate or tilted. The in-plane structures are usually centred rectangular with the chains in the trans conformation and packed in a herringbone pattern. Multilayer films of the acids show only a single-step order-disorder transition at the melting point. This temperature tends to rise as the number of layers increases. Complex changes occur when multilayer films of the salts are heated. Disorder of the chains begins at low temperatures but the arrangement of the head groups does not alter until the melting temperature is reached. Slow heating to a temperature just below the melting temperature gives, with some salts, a radical change in phase. The lamellar structure disappears and a new phase consisting of cylindrical rods lying parallel to the substrate surface and stacked in a hexagonal pattern is formed. In each rod the cations are aligned along the central axis surrounded by the disordered acyl chains. PMID- 11392354 TI - The state of the serotonin transporter protein in the platelets of patients with somatoform [correction of somatiform] disorders. AB - The role of the serotonin transporter protein (STP) in the development of somatoform [corrected] disorders was addressed in a correlational study of the levels of immunoreactive STP (IR-STP) using site-specific antibodies against the least conserved (among a group of other cotransporters) epitope at the C-terminal of STP and the level of anxiety symptoms in patients with somatoform [corrected] disorders. A total of 22 patients were studied, with DSM-IV diagnoses of somatoform [corrected] disorders, along with 32 mentally healthy subjects of comparable age and sex. Immunoblotting of IR-STP from patients from healthy donors produced a diffuse band between 68 and 105 kDal and a clear narrow band at 43 kDal. The 43-kDal IR-STP protein was almost completely absent from most patients, as compared with the levels of this protein in healthy donors. This result suggests an abnormality of STP processing or, perhaps, alternative splicing of the gene encoding STP in patients with somatoform [corrected] disorders, and this appears to reflect the dysfunction in serotoninergic transmission in the CNS in these patients. PMID- 11392357 TI - Modern physicochemical research on Langmuir monolayers. AB - Recent developments in characterising Langmuir monolayers of a variety of film forming materials and employing several physicochemical techniques are reviewed. The extension of the LB method to non-amphiphilic substances, especially macromolecular systems, has increased the need of a thorough understanding of Langmuir film properties, which requires characterising techniques that provide complementary information. Since there is vast literature in the subject, only selected examples are given of results that illustrate the potential of the techniques discussed. PMID- 11392358 TI - On the role of oil-soluble initiators in the radical polymerization of micellar systems. AB - Polymerization in micellar systems is a technique which allows the preparation of ultrafine as well as coarse latex particles. This article presents a review of the current literature in the field of radical polymerization of classical monomers in micellar systems initiated by oil-soluble initiators. Besides a short introduction to some of the kinetic aspects of emulsion polymerization initiated by water-soluble initiators, we mainly focus on the kinetics and the mechanism of radical polymerization in o/w and w/o micellar systems initiated by classical oil soluble initiators. The initiation of emulsion polymerization of an unsaturated monomer (styrene, butyl acrylate,...) by a water-soluble initiator (ammonium peroxodisulfate) is well understood. It starts in the aqueous phase and the initiating radicals enter the monomer-swollen micelle. The formed oligomeric radicals are surface active and increase the colloidal stability of the disperse system. Besides, the charged initiating radicals might experience the energetic barrier when entering the charged particle surface. The locus of initiation with oil-soluble initiators is more complex. It can partition between the aqueous phase and the oil-phase. Besides, the surface-active oil-soluble initiator can penetrate into the interfacial layer. The dissolved oil-soluble initiator in the monomer droplet can experience the cage effect. The small fraction of the oil soluble initiator dissolved in the aqueous phase takes part in the formation of radicals. The oligomeric radicals formed are uncharged and therefore, they do not experience the energetic barrier when entering the polymer particles. We summarize and discuss the experimental data of radical polymerization of monomers initiated by oil-soluble initiators in terms of partitioning an initiator among the different domains of the multiphase system. The inhibitor approach is used to model the formation of radicals and their history during the polymerization. The nature of the interfacial layer and the type of oil-soluble initiator including the surface active ones are related to the kinetic and colloidal parameters. The emulsifier type and reaction conditions in the polymerization are summarized and discussed. PMID- 11392359 TI - Children with difficult asthma: a practical approach. AB - Many open studies investigating the effects of innovative treatments for steroid dependent asthma demonstrate some benefit. This is also true of the majority of placebo arms in placebo-controlled trials. This suggests that children with difficult asthma benefit from the high level of input that is typically provided in clinical trials, with or without additional medication. Such intensive management of patients, with the emphasis on establishing the diagnosis, improving adherence, and identifying provoking factors, is the key to optimizing asthma control for these children. For patients with genuinely severe asthma, despite high doses of conventional treatment, a greater understanding of the pathological basis of persistent symptoms is needed. Identification of different pathological subtypes of severe asthma should allow for more rational prescribing of asthma therapy, as well as the design of further trials of potential steroid sparing treatments. PMID- 11392360 TI - Quality of life in asthma: a comparison of community and hospital asthma patients. AB - This study compares the quality of life of a community sample of people with asthma in South Australia, using population norms, people suffering from other chronic diseases, and a sample of asthma patients from two hospital clinics. A representative population survey was performed by trained interviewers in spring 1995 of 3001 respondents aged > or = 15 years. A physician's diagnosis of current asthma was reported by 299 (9.9%). The hospital clinic sample had a physician's diagnosis and lung function evidence of asthma (n = 293). All completed the SF-36 health survey. Standardized SF-36 scores, adjusted for age, sex, and social class, were significantly lower for respondents with asthma, compared with population norms, across all subscales of the SF-36 (p < 0.05). Physical component summary (PCS) and mental component summary (MCS) scores were not significantly different in people in the community sample with asthma from scores in people with diabetes and arthritis. PCS and MCS scores did not differ for those with similar symptom frequency in the community and hospital asthma samples, except that hospital patients with frequent symptoms had significantly lower MCS scores (p < 0.01). Asthma has a major impact on the health-related quality of life in the community, comparable to other chronic diseases. The SF-36 performs uniformly in asthma in different situations. PMID- 11392361 TI - Severe acute asthma with bowel infarction and pneumatosis intestinalis in a young adult. AB - A young patient presented with a small bowel infarction with pneumatosis intestinalis in the early course of life-threatening severe acute asthma. Low cardiac output with severe congestive right heart failure combined with the use of high doses of epinephrine to reverse the near-fatal bronchospasm probably contributed to this previously unreported complication. The presence of gas collections in the submucosal space was possibly the consequence of diffuse small bowel mucosal disruption. Early recognition of this unusual complication is of major importance to ensure appropriate therapeutic management. PMID- 11392362 TI - Plasma adrenomedullin levels in asthmatic patients. AB - Adrenomedullin (ADM) is a newly discovered endogenous vasorelaxing peptide isolated from pheochromocytoma. Some experimental studies suggest that ADM plays a role in asthma. The purposes of the present study were to assess the plasma ADM levels in adults with mild to severe asthma and controls and to correlate those with the findings on lung function test results and other clinical indices. We recruited 16 mild, 10 moderate, and 11 severely asthmatic patients and 12 healthy controls. We measured the plasma concentrations of ADM in patients with asthma and in healthy subjects using RIA. We assessed FEV1, FEV1 predicted %, FEV1/FVC, symptom score, IgE, ECP, and morning and evening peak expiratory flow measurements. There was no significant difference between the asthmatic and the control group ADM levels, which were 26.3 +/- 24.2 pg/mL and 22.9 +/- 17.6 pg/mL, respectively. Furthermore, plasma ADM levels increased as the severity of the disease increased in asthmatic patients (20.7 +/- 14.4 pg/mL in mild, 25.2 +/- 24.3 pg/mL in moderate, and 35.5 +/- 33.6 pg/mL in severe asthmatics), although they did not result in any statistical significance. However, the plasma ADM levels correlated negatively with the FEV1 levels in the asthmatic group (p < 0.02, r = -0.37). Peripheral blood eosinophilia, IgE, and ECP levels did not correlate with plasma ADM levels. These results suggest that the measurement of ADM concentration in plasma will not be of diagnostic use in asthma, but may be a reflection of the severity of asthma. PMID- 11392363 TI - A comparison of the efficacy and safety of a half dose of fluticasone propionate with beclamethasone dipropionate and budesonide in childhood asthma. AB - This study was carried out in an attempt to compare the efficacy and safety of fluticasone propionate (FP) at the half dose of budesonide (BUD) and beclamethasone dipropionate (BD) in childhood asthma. Ninety-six children with moderate to severe asthma (9.6 +/- 2.17 years) whose asthma was already controlled on BUD (n = 52) or BD (n = 44) were recruited into the study. In the first part of the study (the first 12 weeks) each group was followed with three weekly lung function measurements, daily diary records, and peak expiratory flow (PEF) measurements on the initial medication. At the end of 6 weeks, drugs were switched to a half dose of FP, and the subjects were followed for another 6 weeks. Blood samples were obtained for osteocalcin and plasma cortisol levels after each treatment period. In the second part of the study, 50 patients continued to take FP at the half dose of BUD or BD for another 30 weeks. Clinic visits, including lung function and PEF measurements, were conducted every 10 weeks. After 6 weeks of FP treatment, there was a small but statistically significant decrease in FEV1 and FEF(25-75) in both groups (BUD and BD) without any significant obstruction. These mild changes in lung function measurements continued during long-term follow-up. However, there was no statistically significant further decrease in any lung function parameters while receiving FP (visits 3-8) (coefficient = -0.00751 L/day, p = 0.39 for FEF(25-75) and coefficient = -0.00910 L/sec/day, p = 0.055 for FEV1). There were no significant changes in the morning and evening PEF measurements and diurnal PEF variations after 6 weeks of treatment with FP compared with BUD and BD treatments. There were no significant changes in basal cortisol and osteocalcin levels before or after 6 weeks of FP treatment (p > 0.05). The present study concluded that, although FP at the half dose of BUD or BD seems to maintain reasonable control of the disease symptoms, a mild but significant and persistent decrease in lung function parameters may indicate that FP may not be twice as potent as BUD or BD in childhood asthma by evaluation of lung functions. This conclusion must be further verified with long-term studies. PMID- 11392364 TI - Household smoking and bronchial hyperresponsiveness in children with asthma. AB - This study investigated whether household environmental tobacco smoke (ETS) exposure is associated with increased bronchial hyperresponsiveness (BHR) in children with asthma. Two hundred forty-nine children, ages 7-11 years, sampled from a larger group with reported asthma or multiple asthma symptoms identified in a community survey in Cape Town, underwent histamine challenge testing and had urinary cotinine measured. Parents were interviewed for information on smoking habits and a variety of covariates. Children with asthma whose mothers smoked had a lower frequency of BHR than asthmatic children of nonsmoking mothers, particularly if the mother smoked > or = 15 cigarettes daily. BHR was also less common among children sharing a house with four or more smokers vs. fewer or none. BHR was unrelated to paternal smoking. In contrast, FEV1 was lower among children whose mothers currently smoked. The findings do not support a mechanism whereby ETS exposure aggravates existing childhood asthma by increasing BHR. This association may be masked, however, by the degree to which mothers of asthmatic children adjust their smoking. The results are consistent with an adverse effect of maternal smoking on lung function in asthmatic children. PMID- 11392365 TI - Rural school nurses' asthma education needs. AB - School nurses play an important role in identifying children with asthma and providing care during school hours. Educational programs designed to improve nurses' asthma knowledge and practices have concentrated on urban settings. The purpose of this investigation was to determine asthma-related practices and educational needs of rural school nurses. A survey about asthma was mailed to school nurses in all counties of the state of Maryland and in Washington, D.C. Responses were compared between rural Maryland counties and counties from the remainder of Maryland and Washington, D.C. The survey addressed attitudes and beliefs, function and roles, medication administration, and educational needs about asthma. We found that rural nurses used peak flow meters less often to assess and monitor asthma, requested fewer referrals for asthma, had fewer interactions with health room assistants, and had reduced access to asthma educational resources. Also, they provided less asthma education in the schools than other school nurses. These results suggest a need for comprehensive asthma educational programs in rural areas that are based on national guidelines, and that address the unique needs of rural school nurses. These programs should also emphasize the need for open communication between rural school nurses, health room assistants, primary care providers, and parents/caregivers. PMID- 11392366 TI - Asthma patient education: current utilization in pulmonary training programs. AB - The national guidelines for the diagnosis and management of asthma published in April of 1997 emphasized patient education in asthma management. It is unclear how often patient education is included in asthma management clinics. We sought to determine how often education programs are available by surveying teaching hospitals with training programs in pulmonary and critical care medicine. Using this survey, we also determined the reason programs are not offered and whether computer resources are routinely available to utilize computer-delivered patient educational materials. We sent mail questionnaires to 163 training programs in the United States. We had a response rate of 72% (117 of 163). Of the 117 programs responding, 75 (64%) reported having a formal asthma patient education program. Most (72%) were in university teaching hospitals. A majority of respondents (84%) believed that compensation for their efforts was inadequate, and those hospitals with no formal asthma education program reported that financial cost and time requirements were the primary reasons for not having such a program. Despite the fact that many programs did not have a patient education component, 96% (72 of 75) of respondents with an educational program viewed patient education as an effective patient self-management tool. Of all programs surveyed, 85% reported they would use a high-quality computer-based asthma education program if one was available. Implementation of such a program is feasible, with 69% of programs surveyed having a personal computer in their clinic and 60% having Internet access. We conclude that most training directors believe that patient asthma education is important and effective; however, cost and time issues remain barriers to its implementation. Computer-based educational programs delivered over the Internet are feasible, could address some of these limitations, and are acceptable to most programs. PMID- 11392367 TI - Can asthma education improve clinical outcomes? An evaluation of a pediatric asthma education program. AB - Asthma is a common, costly, and chronic disease that has a significant impact on children, their families, and the health care system. This study investigated whether formal asthma education can reduce asthma severity and morbidity and further questioned whether the method of education is an important factor in this process. Study subjects were recruited from the emergency department of the Royal Children's Hospital, Melbourne (RCHM), and controls were recruited from the neighboring Western Hospital, Sunshine (WHS). Subject parents were randomized to receive face-to-face education, group education, or home-based video education. Controls had similar medical treatment but received no formal asthma education. Subject and control parents were asked to complete questionnaires before the intervention and at 1, 6, and 12 months. These questionnaires measured demographics, the levels of parent asthma knowledge, anxiety, child asthma severity, and morbidity. Results showed that any method of education increased parent asthma knowledge and immediately decreased their anxiety and child asthma morbidity scores. Despite this, asthma severity was reduced only when knowledge was imparted in an interactive, face-to-face setting. When this happened, asthma severity scores were significantly reduced in both the short- and long-term. These findings call into question mass asthma education campaigns and have important implications for the design of future asthma education programs. PMID- 11392368 TI - The relationship between the quality of life of children with asthma and family functioning. AB - The quality of life of 84 children with asthma, aged 7-12 years, and their family functioning was assessed using standard questionnaires. A significant relationship was found between children's reports of their quality of life and several key dimensions of family functioning. The results suggest that independent of their frequency, the extent to which asthma symptoms upset and bother children varies depending on the level of the functioning of the children's families. Treatment approaches designed to improve family functioning may reduce the extent to which children are bothered by their asthma symptoms and thus improve their quality of life. PMID- 11392369 TI - Effects of phthalate esters on the developing reproductive tract of male rats. AB - Phthalate esters are a large group of chemical agents used predominantly as plasticizers and solvents. Certain members of this chemical class have been shown to cause reproductive and developmental toxicity. Recent attention has focused on the potential of these agents to interfere with male reproductive development through a postulated antiandrogenic mechanism. Observations have focused on di-n butyl phthalate (DBP), di-(2-ethylhexyl) phthalate (DEHP) and butyl benzylphthalate, with most information relating to dose-response relationships obtained for DBP. Neither DBP, DEHP nor their major metabolites interacted with human or rodent androgen receptors (AR) in transcriptional activation assays. DBP was administered during the critical window of development of the male reproductive system, after which the resulting offspring were examined until adulthood. DBP elicited marked effects on the developing male reproductive tract, including malformations of the epididymis and vas deferens, and hypospadias. Retention of thoracic nipples/areolae and reductions in anogenital distance were also noted. Surprisingly, Leydig cell adenomas were induced in some male offspring at 100 days of age. All these events occurred in the absence of any toxicity in the pregnant dam. Examination of testes from fetal rats indicated markedly reduced testosterone levels and increased Leydig cell numbers after DBP administration to the dams. Leydig cells were positive for AR and 3 betahydroxysteroid dehydrogenase. PMID- 11392370 TI - Neonatal exposure to potent and environmental oestrogens and abnormalities of the male reproductive system in the rat: evidence for importance of the androgen oestrogen balance and assessment of the relevance to man. AB - The effects on reproductive tract development in male rats, of neonatal exposure to potent (reference) oestrogens, diethylstilboestrol (DES) and ethinyl oestradiol (EE), with those of two environmental oestrogens, octylphenol and hisphenol A were systematically compared. Other treatments, such as administration of a gonadotrophin-releasing hormone antagonist (GnRHa) or the anti-oestrogen tamoxifen or the anti-androgen flutamide, were used to aid interpretation of the pathways involved. All treatments were administered in the neonatal period before onset of puberty. The cellular sites of expression of androgen receptors (AR) and of oestrogen receptor-alpha (ERalpha) and ERbeta were also established throughout development of the reproductive system. The main findings were as follows: (i) all cell types that express AR also express one or both ERs at all stages of development; (ii) Sertoli cell expression of ERbeta occurs considerably earlier in development than does expression of AR; (iii) most germ cells, including fetal gonocytes, express ERbeta but not AR; (iv) treatment with high, but not low, doses of potent oestrogens such as DES and EE, induces widespread structural and cellular abnormalities of the testis and reproductive tract before puberty; (v) the latter changes are associated with loss of immunoexpression of AR in all affected tissues and a reduction in Leydig cell volume per testis; (vi) none of the effects in (iv) and (v) can be duplicated by treating with high-dose octylphenol or bisphenol A; (vi) none of the reproductive tract changes in (iv) and (v) can be induced by simply suppressing androgen production (GnRHa treatment) or action (flutamide treatment); and (vii) the adverse changes induced by high-dose DES (iv and v) can be largely prevented by co-administration of testosterone. Thus, it is suggested that many of the adverse changes to the testis and reproductive tract induced by exposure to oestrogens result from a combination of high oestrogen and low androgen action. High oestrogen action or low androgen action on their own are unable to induce the same changes. PMID- 11392371 TI - Effects of environmental antiandrogens on reproductive development in experimental animals. AB - Chemicals that act as androgen receptor (AR) agonists and antagonists or inhibit fetal steroidogenesis can induce reproductive malformations in humans and laboratory animals. Several environmental chemicals disrupt development in rats and/or rabbits at fetal concentrations at, or near, exposure levels seen in some segments of the human population. In rats, fetal tissues concentrations of 10-20 p.p.m. of the DDT metabolite, p,p'-DDE, are correlated with reproductive abnormalities in male offspring. These concentrations are similar to those measured in first-trimester human fetal tissues in the late 1960s. The pesticides vinclozolin, procymidone, linuron and DDT are AR antagonists. They reduce male rat anogenital distance, and induce areolas at relatively low dosages. Hypospadias, agenesis of the sex accessory tissues and retained nipples are seen in the middle dosages, while undescended testes and epididymal agenesis are seen in the highest doses. Phthalate esters (PE) inhibit testosterone synthesis during fetal life, but do not appear to be AR antagonists. Prenatal administration of a single low dose of dioxin (50-1,000 ng TCDD/kg) alters the differentiation of androgen-dependent tissues at p.p.t. concentrations, but the mechanism of action likely involves interaction with a hormone-like nuclear transcription factor, the hormone-like receptor AhR, rather than AR. p,p'-DDT and p,p'-DDE, vinclozolin and di-n-butyl phthalate affect reproductive function in rabbits when administered during prenatal and/or neonatal life. Cryptorchidism and carcinoma in situ-like (CIS) testicular lesions were seen in male rabbits treated during development with p,p'-DDT or p,p'-DDE. Extrapolation of effects from rodents to humans would be enhanced if future studies incorporate determination of tissue concentrations of the active metabolites. Knowledge of the tissue concentrations of the active toxicants also would provide an important link to in-vitro studies, which provide more useful mechanistic information when they are executed at relevant concentrations. PMID- 11392372 TI - A re-examination of variation associated with environmentally stressed organisms. AB - Variation is an essential feature of biological systems. Populations adapt to dynamic environments, in part, because of this variation. In this review, we re examine phenotypic variation, especially in organisms living in polluted environments. A recent goal of ecotoxicology is to understand the sublethal effects of exposure to pollutants, e.g. responses to endocrine-disrupting contaminants. While variation is an inherent quality of organisms, variance is a statistical measure of the variation of a trait. Increased variance has been associated with organisms living at the perimeter of a population's range, introduced into novel environments, or exposed to pollution. Some researchers have proposed increased phenotypic variance in exposed populations as an evolutionary mechanism, and others have suggested its use as a biomarker. While we agree that variance often increases in the exposed population, we also recognize that the opposite phenomenon occurs. That is, variance can decrease from exposure to pollution. Altered variance in the exposed population-leading to heteroscedasticity-could result in erroneous conclusions (Type II errors). We suggest that exposure to endocrine-disrupting contaminants could influence the health of populations in ways that are not always represented by measures of central tendency, and that variance and distribution should also be examined in environmentally stressed wildlife. PMID- 11392373 TI - Genotoxicity of the steroidal oestrogens oestrone and oestradiol: possible mechanism of uterine and mammary cancer development. AB - Oestrogens, including the natural hormones oestrone and oestradiol, induce various tumours in laboratory animals and have been recognized to be carcinogens in humans, raising the risk for breast and uterine cancer. As part of the search for the mechanism of hormone-induced carcinogenesis, various types of DNA damage have been detected which have been induced by oestrogens in cell-free systems, in cells in culture, or in vivo. Nevertheless, oestrogens have been postulated to act only as promoters of mammary carcinogenesis by receptor-mediated growth stimulation without consideration of their genotoxicity because these hormones failed to induce mutations in commonly used assays. More recently, oestradiol induced numerical chromosomal changes (aneuploidy) and structural chromosomal aberrations have been detected in cells in culture and in hamster kidney, a target of oestrogen-induced cancer. In this animal model, oestradiol generates c myc gene amplification and microsatellite instability. Mutations of the hprt gene have been induced by oestradiol in V79 cells and by catecholoestrogen metabolites in Syrian hamster embryo cells. Sequencing of this gene isolated from V79 mutant clones revealed point mutations and deletions. It is concluded that oestradiol plays a dual role as mutagen/carcinogen and as growth-stimulating hormone in the induction of tumours. PMID- 11392374 TI - Trends in the incidence of cryptorchidism and hypospadias, and methodological limitations of registry-based data. AB - Cryptorchidism and hypospadias share possible risk factors, such as intrauterine growth retardation. According to the data collected by the International Clearinghouse for Birth Defects Monitoring Systems (ICBDMS), apparently increasing trends in the incidence of hypospadias were found in Sweden during the 1960s, and in Norway, Denmark, England and Hungary during the 1970s. In Norway and Denmark, the increase continued in the 1980s, while in the USA it has continued from the 1970s to the 1990s. Finland has shown a lower reported rate of hypospadias than other Nordic countries. However, it is difficult to make comparisons between countries because of variable inclusion criteria. Furthermore, the reliability of the data depends on correct ascertainment and reporting of the cases. The ICBDMS has also collected data on cryptorchidism, but these appear to be unreliable because of a discrepancy with the data from cohort studies. According to two comparable English studies, the incidence of cryptorchidism in full-term boys approximately doubled between the 1950s and the 1980s. Regionally there are large differences: e.g. in Finland the incidence of cryptorchidism is clearly lower than in Denmark. Regional and temporal trends may help to identify environmental factors that might be associated with these disorders. PMID- 11392375 TI - Trends in pubertal development in Europe. AB - The secular changes in growth and maturation can be seen as indicators of socio economic and health status. In most European countries the age of onset of puberty and of menarcheal age has been decreasing during the past few decades. The duration of puberty seems also to decrease, though few studies provide sufficient data to support this postulation. The four Dutch nationwide growth surveys are useful examples assessing the secular trend in pubertal development over the past 45 years. Genetic and environmental factors contribute to the secular changes. Environmental factors seem to be the most important. Recently, attention has been given to substances with oestrogen-like actions that are present in nutrients. The possible role of these substances in growth and maturation is discussed. PMID- 11392376 TI - Pathogenesis and epidemiology of precocious puberty. Effects of exogenous oestrogens. AB - Precocious puberty is generally defined as the appearance of secondary sex characteristics before age 8 years in girls (or menarche before age 9 years) and before 9 years in boys. The overall incidence of sexual precocity is estimated to be 1:5,000 to 1:10,000 children. The female-to-male ratio is approximately 10:1. In addition to the psychosocial disturbances associated with precocious puberty, the premature pubertal growth spurt (with less time for prepubertal growth) and the accelerated bone maturation result in reduced adult height. Precocious puberty may be gonadotrophin-dependent [i.e. of central origin with premature activation of the gonadotrophin-releasing hormone (GnRH) pulse generator] or gonadotrophin-independent (i.e. peripheral where the GnRH pulse generator is suppressed). This can be determined by GnRH testing. The pathophysiology is the basis for different diagnostic and therapeutic strategies, i.e. in the first case a stimulated LH/FSH ratio >1 and suppressive treatment with GnRH agonists (e.g. in hypothalamic hamartoma), and in the second decreased gonadotrophins and removal or suppression of the endogenous or exogenous sex steroid source (e.g. congenital adrenal hyperplasia). While several cases of gonadotrophin-independent precocious puberty due to oestrogen exposure via the transdermal, oral, or inhalative route have been reported, no case is known with the development of subsequent secondary central precocious puberty. Food contamination with oestrogens is theoretically possible, but would most probably be sporadic and, thus, would not lead to precocious puberty. As steroid hormones in meat production are banned in the European Union, no data on the impact of environmental oestrogenic substances on human maturation are currently available. In conclusion, the risk for children to develop precocious puberty through exposure to oestrogens (or androgens) in the environment or in food is very low. Nevertheless, studies of the effects of defined environmental oestrogenic substances on the human reproductive system and on pubertal development are warranted. PMID- 11392377 TI - The effects of oestrogens on linear bone growth. AB - Regulation of linear bone growth in children and adolescents comprises a complex interaction of hormones and growth factors. Growth hormone (GH) is considered to be the key hormone regulator of linear growth in childhood. The pubertal increase in growth velocity associated with GH has traditionally been attributed to testicular androgen secretion in boys, and to oestrogens or adrenal androgen secretion in girls. Research data indicating that oestrogen may be the principal hormone stimulating the pubertal growth spurt in boys as well as girls is reviewed. Such an action is mediated by oestrogen receptors (ER-alpha and ER beta) in the human growth plate, and polymorphisms in the ER gene may influence adult height in healthy subjects. Prepubertal oestradiol concentrations are significantly higher in girls than in boys, explaining sex-related differences in pubertal onset. Men with a disruptive mutation in the ER gene (oestrogen resistance) or in the CYP19 gene (aromatase deficiency) who have no pubertal growth spurt and continue to grow into adulthood due to lack of epiphyseal fusion supports this notion. Furthermore, phenotypic females with complete androgen insensitivity syndrome have a normal female growth spurt despite lack of androgen action. Oestrogens may also influence linear bone growth indirectly via modulation of the GH-insulin-like growth factor-I (IGF-I) axis. Thus, ER blockade diminishes endogenous GH secretion, androgen receptor (AR) blockade increases GH secretion in peripubertal boys, and non-aromatizable androgens [oxandrolone or dihydrotestosterone (DHT)] have no effect on GH secretion. Treatment with aromatase inhibitors reduces circulating IGF-I concentrations in healthy males, and reduces growth in boys with testotoxicosis. Taken together, these findings suggest that oestrogens may, in addition to their direct effects, stimulate GH secretion and thereby increase circulating IGF-I, which in turn may stimulate growth. Thus, oestrogens have important biphasic actions on longitudinal growth in boys as well as in girls. Very low levels of oestrogens may stimulate bone growth without affecting sexual maturation directly at the growth plate as well as through stimulation of the GH-IGF axis, which in turn may stimulate growth. Conversely, higher levels of oestrogens stimulate secondary sexual characteristics and epiphyseal fusion. PMID- 11392378 TI - Disorders linked to insufficient androgen action in male children. AB - Virilization of the external genitalia in the male fetus requires testosterone and dihydrotestosterone (DHT), which is formed from testosterone by the action of the enzyme, 5alpha-reductase type 2 (5alphaR-2). Mediation of the effects of both testosterone and DHT requires a functional androgen receptor (AR) located in the cytoplasmic compartment of target cells. DHT (or testosterone) binding induces a conformational change which facilitates AR nuclear transport, phosphorylation and dimerization, ultimately regulating of the rate of transcription of androgen dependent genes. Any event which impairs DHT formation (mutation within the 5alphaR-2 gene or 5alphaR-2 inhibitors) or normal function of the AR (mutation in the AR gene, antiandrogens) may result in insufficient androgen action in the male fetus and in subsequent undervirilization in the newborn. Hypospadias may be due to a defect in androgen action due to mutation of the 5alphaR-2 or of the AR gene. Mutation of unidentified genes is likely to underlie this displacement of the urethral meatus from the tip to the ventral side of the phallus. An aetiological role for environmental chemical products has been postulated, since ethnic as well as geographical differences in the incidence of hypospadias have been noted. Increasing evidence has been gathered indicating that widely used industrial and agricultural chemicals have deleterious effects on normal male sexual differentiation. Cryptorchidism and micropenis may represent an intersex phenotype, even if they are isolated. Aetiological factors include 5alphaR-2 gene mutation, AR gene mutation or environmental hormonal disruptors. In conclusion, several phenotypes have been attributed to insufficient androgen action during fetal life. Whereas mutations in the 5alphaR-2 gene and AR gene are natural, attention should be focused on environmental endocrine disruptors that are able to mimic steroid 5alpha-reductase deficiency or partial androgen insensitivity syndrome. PMID- 11392379 TI - The impact of endocrine disrupters on the female reproductive system. AB - Over the last decades, many tonnes of man-made chemicals have been produced and released into the environment. Many of these chemical substances have the ability to modulate the action of hormones and are called endocrine disrupters. Cell receptors that have been pure receptors for thousands of years have (due to industrialization), become susceptible to the action of exogenous chemicals. The balance of the endocrine system is very important in the human body especially in females because the menstrual cycle and fertility are very sensitive to hormone imbalances. This review considers the mode of exposure and action of endocrine disrupters and focuses on their impact on the female reproductive system, including female hormone concentrations, menstrual cycle, fertility, spontaneous abortion and the development of endometriosis. An attempt is made to elucidate the impact of endocrine disrupters on the female reproductive system, while admitting that most scientific data come from experimental animals and the conclusions cannot be applied to humans easily. The aim is to present available information, highlighting the impact of endocrine disrupters on the female reproductive system, in order to stimulate reevaluation in identifying hormone disorders. PMID- 11392380 TI - Human health effects of dioxins: cancer, reproductive and endocrine system effects. AB - Polychlorinated dioxins, furans and polychlorinated benzene constitute a family of toxic persistent environmental pollutants. In Europe, environmental concentrations increased slowly throughout this century until the late 1980s. Dioxins have been shown to be carcinogenic in animals and humans. In humans, excess risks were observed for all cancers, without any specific cancer predominating. In specific cohorts, excess risks were observed for reproductive cancers (breast female, endometrium, breast male, testis) but, overall, the pattern is inconsistent. In animals, endocrine, reproductive and developmental effects are among the most sensitive to dioxin exposure. Decreased sperm counts in rats and endometriosis in rhesus monkeys occur at concentrations 10 times higher than current human exposure. In humans, results are inconsistent regarding changes in concentrations of reproductive hormones. A modification of the sex ratio at birth was described in Seveso. There exist no data on effects such as endometriosis or time-to-pregnancy. Small alterations in thyroid function have occasionally been found. Increased risk for diabetes was seen in Seveso and a herbicide applicators cohort but, overall, results were inconsistent. Experimental data indicate that endocrine and reproductive effects should be among the most sensitive effects in both animals and humans. Epidemiological studies have evaluated only a few of these effects. PMID- 11392382 TI - Perspective: reverse evolution. AB - For some time, the reversibility of evolution was primarily discussed in terms of comparative patterns. Only recently has this problem been studied using experimental evolution over shorter evolutionary time frames. This has raised questions of definition, experimental procedure, and the hypotheses being tested. Experimental evolution has provided evidence for multiple population genetic mechanisms in reverse evolution, including pleiotropy and mutation accumulation. It has also pointed to genetic factors that might prevent reverse evolution, such as a lack of genetic variability, epistasis, and differential genotype-by environment interactions. The main focus of this perspective is on laboratory studies and their relevance to the genetics of reverse evolution. We discuss reverse evolution experiments with Drosophila, bacterial, and viral populations. Field studies of the reverse evolution of melanism in the peppered moth are also reviewed. PMID- 11392381 TI - Possible health impact of animal oestrogens in food. AB - Oestrogens govern reproductive functions in vertebrates, and are present in all animal tissues. The theoretical maximum daily intake (TMDI) of oestradiol-17beta by consumption of cattle meat is calculated to be 4.3 ng. Following the use of oestradiol-containing growth-promoting agents, TMDI is increased by a factor of 4.6 to 20 ng oestradiol-17beta, assuming that single dosage and 'good animal husbandry' are observed. Pork and poultry probably contain similar amounts of oestrogens as untreated cattle. The mean concentration of oestradiol-17beta in whole milk is estimated at 6.4 pg/ml. Scarce data available on eggs report up to 200 pg/g oestradiol-17beta. The risk evaluation of oestrogenic growth-promoting agents is limited by analytical uncertainties. Residues of oestradiol-17alpha and the importance of oestrogen conjugates are widely unknown. The performance of mass spectrometry still needs to be improved for confirmation of oestrogen concentrations in most food. At present, the potential relevance of oestradiol acyl esters, the actual daily production rate of oestradiol in prepubertal children, and the role of oestradiol metabolites in cancer are obscure. The presence of different cytoplasmic oestrogen receptor subtypes and potential oestradiol effects in non-reproductive functions require further examination. PMID- 11392383 TI - Inferring speciation rates from phylogenies. AB - It is possible to estimate the rate of diversification of clades from phylogenies with a temporal dimension. First, I present several methods for constructing confidence intervals for the speciation rate under the simple assumption of a pure birth process. I discuss the relationships among these methods in the hope of clarifying some fundamental theory in this area. Their performances are compared in a simulation study and one is recommended for use as a result. A variety of other questions that may, in fact, be the questions of primary interest (e.g., Has the rate of cladogenesis been declining?) are then recast as biological variants of the purely statistical question-Is the birth process model appropriate for my data? Seen in this way, a preexisting arsenal of statistical techniques is opened up for use in this area: in particular, techniques developed for the analysis of Poisson processes and the analysis of survival data. These two approaches start from different representations of the data--the branch lengths in the tree--and I explicitly relate the two. Aiming for a synoptic account of useful theory in this area, I briefly discuss some important results from the analysis of two distinct birth-death processes: the one introduced into this area by Hey (1992) is refitted with some powerful statistical tools. PMID- 11392384 TI - Genotype-by-environment interaction and the fitness of plant hybrids in the wild. AB - Natural hybrid zones between related species illustrate processes that contribute to genetic differentiation and species formation. A common viewpoint is that hybrids are essentially unfit, but they exist in a stable tension zone where selection against them is balanced by gene flow between the parent species. An alternative idea is that selection depends on the environment, for example, by favoring opposite traits in the two parental habitats or favoring hybrids within a bounded region. To determine whether selection of hybrids is environment dependent, we crossed plants of naturally hybridizing Ipomopsis aggregata and I. tenuituba in the Colorado Rocky Mountains and reciprocally planted the seed offspring into a suite of natural environments across the hybrid zone. All types of crosses produced similar numbers and weights of seeds. However, survival of the offspring after 5 years differed markedly among cross types. On average, the F1 hybrids had survival and growth rates as high as the average for their parents. But hybrid survival depended strongly on the direction of a cross, that is, on which species served as the maternal parent. This fitness difference between reciprocal hybrids appeared only in the parental environments, suggesting cytonuclear gene interactions that are environment specific. These results indicate that complex genotype-by-environment interactions can contribute to the evolutionary outcome of hybridization. PMID- 11392385 TI - Evolutionary rates and species diversity in flowering plants. AB - Genetic change is a necessary component of speciation, but the relationship between rates of speciation and molecular evolution remains unclear. We use recent phylogenetic data to demonstrate a positive relationship between species numbers and the rate of neutral molecular evolution in flowering plants (in both plastid and nuclear genes). Rates of protein and morphological evolution also correlate with the neutral substitution rate, but not with species numbers. Our findings reveal a link between the rate of neutral molecular change within populations and the evolution of species diversity. PMID- 11392386 TI - Low intraspecific variation for genomic isolation between hybridizing sunflower species. AB - Barriers to gene flow between species result from selection against foreign linkage blocks in hybrids. When the geographic ranges of taxa meet at multiple locations, the opportunity exists for variation in the genetic architecture of isolating barriers. Hybrid zones between two sunflower species (Helianthus annuus and H. petiolaris) in Nebraska and California exhibited remarkably similar patterns of introgression of mapped molecular markers. Congruence among hybrid zones may result from limited intraspecific variation at loci contributing to isolation and from similar selective effects of alleles in the heterospecific genetic background. The observed consistency of introgression patterns across distantly separated hybrid zones suggests that intrinsic forces predominate in determining hybrid zone dynamics and boundaries between these sunflower species. PMID- 11392387 TI - Adaptive divergence in plasticity in natural populations of Impatiens capensis and its consequences for performance in novel habitats. AB - We tested for adaptive differentiation between two natural populations of Impatiens capensis from sites known to differ in selection on plasticity to density. We also determined the degree to which plasticity to density within a site was correlated with plastic responses of experimental immigrants to foreign sites. Inbred lines, derived from natural populations in an open-canopy site and a woodland site, were planted reciprocally in both original sites at naturally occurring high densities and at low density. The density manipulation represents environmental variation typically experienced within the site of a given population, and the transplant manipulation represents environmental differences between sites of different populations. Internode elongation, meristem allocation, leaf length, flowering date, and total lifetime fitness were measured. Genotypes originating in the open site, where selection favored plasticity of first internode length and flowering time (Donohue et al. 2000a), were more plastic in those characters than genotypes originating from the woodland site, where plasticity was maladaptive. Therefore, these two populations appear to have responded to divergent selection on plasticity. Plasticity to density strongly resembled plasticity to site differences for many characters, suggesting that similar environmental factors elicit plasticity both to density and to overhead canopy. Thus, plasticity that evolved in response to density variation within a site influenced phenotypic expression in the foreign site. Plastic responses to site caused immigrants from foreign populations to resemble native genotypes more closely. In particular, immigrants from the open site converged toward the selectively favored early-flowering phenotype of native genotypes in the woodland site, thereby reducing potential fitness differences between foreign and native genotypes. However, because genotypes from the woods population were less plastic than genotypes from the sun population, phenotypic differences between populations were greatest in the open site at low density. Therefore, population differences in plasticity can cause genotypes from foreign populations to be more strongly selected against in some environments than in others. However, genetic constraints and limits to plasticity prevented complete convergence of immigrants to the native phenotype in any environment. PMID- 11392388 TI - Chloroplast sharing in the Tasmanian eucalypts. AB - The biogeographic pattern of chloroplast DNA (cpDNA) haplotypes in Eucalyptus on the island of Tasmania is consistent with reticulate evolution, involving at least 12 Tasmanian species from the subgenus Symphyomyrtus. Intraspecific cpDNA polymorphism in 14 of 17 species is coupled with extensive sharing of identical haplotypes across populations of different species in the same geographic area. Haplotype diversity is lowest in central regions of Tasmania formerly occupied by alpine vegetation during glacial intervals and in northern regions that were periodically linked to continental Australia by land bridges. The observed distribution of several cpDNA haplotypes unique to Tasmania coincides with modeled locations of glacial refugia in coastal areas of Tasmania and shows the power of cpDNA in unraveling the complex history of past distributions of Eucalyptus. The results suggest that the model of evolution of the eucalypts should be reassessed to allow for the anastomosing effects of interspecific hybridization and introgression. PMID- 11392389 TI - Tarsal asymmetry, nutritional condition, and survival in water boatmen (Callicorixa Vulnerata). AB - Fluctuating asymmetry (FA) has been used as a measure of developmental stability and may indicate individual phenotypic or genotypic quality. Using water boatmen (Callicorixa vulnerata) from a natural population, we examined the relationship between tarsal FA (tarsal spine number, tarsal length) and indices of body condition in two habitats. We used body weight and residual body weight (controlling for body length) as indices of condition because experimental food deprivation in water boatmen led to a reduction in each. We detected a negative relationship between FA and both indices of condition in two ecologically distinct pond habitats. We predicted this association was due to a negative relationship between FA and competitive feeding ability. Consequently, we examined associations between survival time and tarsal FA in C. vulnerata under resource-limited laboratory conditions. Univariate analyses revealed a negative correlation between survival and tarsal FA in each trait. Inclusion of survival time, body length, gender, tarsal spine number, tarsal length, and measures of FA into multivariate analyses revealed a negative correlation between survival and FA. Individuals with the greatest survival had higher nutritional condition than individuals that succumbed early in the experiment. Asymmetric individuals may suffer a foraging handicap as a result of the use of tarsi in feeding or they may be of poor genetic quality. Our results suggest elevated FA may limit resource acquisition and are consistent with the use of FA as a measure of fitness. PMID- 11392390 TI - Characterization of female preference functions for Drosophila montana courtship song and a test of the temperature coupling hypothesis. AB - Female mate preferences are a major cause of diversity and elaboration in male sexual traits. Here we characterize the shape of female preference functions for pulse length and carrier frequency of the courtship song of Drosophila montana by fitting both parametric and nonparametric functions to the incidence of female receptive gestures to synthetic song. Preference functions for both traits are strongly directional. That for pulse length is linear and favors short pulses, whereas that for carrier frequency is stabilizing in shape, but would exert directional preferences favoring males with high carrier frequency. The preference for carrier frequency has probably evolved under sexual selection, but reasons for the preference for short pulses are less apparent. We also examine the effect of ambient temperature on the carrier frequency of male song and on the preference function for carrier frequency. For many similar acoustic communication systems, temperature coupling, a compensatory effect of temperature on preference functions, is thought to maintain coordination between preferences and signals. However, although the carrier frequency of D. montana song is highly dependent on environmental temperature, there is no temperature coupling of the female preference function. We suggest that temperature coupling may often arise due to a common effect of temperature on song and preference, rather than be an advantageous characteristic whose function is to maintain coordination in temperature-affected communication systems. PMID- 11392391 TI - The Y chromosomes of Drosophila simulans are highly polymorphic for their ability to suppress sex-ratio drive. AB - The sex-ratio trait, known in several species of Drosophila including D. simulans, results from meiotic drive of the X chromosome against the Y. Males that carry a sex-ratio X chromosome produce strongly female-biased progeny. In D. simulans, drive suppressors have evolved on the Y chromosome and on the autosomes. Both the frequency of sex-ratio X and the strength of the total drive suppression (Y-linked and autosomal) vary widely among geographic populations of this worldwide species. We have investigated the pattern of Y-linked drive suppression in six natural populations representative of this variability. Y linked suppressors were found to be a regular component of the suppression, with large differences between populations in the mean level of suppression. These variations did not correspond to differences in frequency of discrete types of Y chromosomes, but to a more or less wide continuum of phenotypes, from nonsuppressor to partial or total suppressor. We concluded that a large diversity of Y-linked suppressor alleles exists in D. simulans and that some populations are highly polymorphic. Our results support the hypothesis that a Y-chromosome polymorphism can be easily maintained by a balance between meiotic drive and the cost of drive suppression. PMID- 11392392 TI - Behavioral differentiation in oviposition activity in Drosophila buzzatii from highland and lowland populations in Argentina: plasticity or thermal adaptation? AB - Highland populations of several Drosophila species in Argentina were active early in the afternoon in the field as opposed to populations from a much warmer lowland site, where flies were mainly active in the early evening prior to sunset. For one of these species, Drosophila buzzatii, we tested for a genetic component of activity differences by carrying out crosses within and between populations and measuring oviposition activity of the progeny in the laboratory. We found that activity in the highland population exceeded that in the lowland one during the midafternoon, whereas activity in the lowland population exceeded that in the highland one prior to the beginning of the dark period. Oviposition activity for the period corresponding to the field observations was regressed on the proportion of the genome derived from the highland population. This variable significantly predicted oviposition activity between 1400 and 1600 and between 2000 and 2200 h. Activity of both reciprocal crosses was intermediate and not significantly different from each other, suggesting that nuclear genetic, rather than cytoplasmic factors contribute to differences in oviposition activity between the populations. Two morphological, one genetic, and one stress resistance trait were also scored to examine whether temperature differences between environments were associated with other differences between populations. Wing length of wild-caught and laboratory-reared flies from the highland population significantly exceeded that in the lowland. Thorax length of laboratory-reared flies from the highland population also significantly exceeded that from the lowland. Chromosomal inversion frequencies differed significantly between the two populations with a fivefold reduction in the frequency of arrangement 2st in the highland as compared to the lowland population. This arrangement is known for its negative dose effect on size, and thus, the highland population has experienced a genetic change, perhaps as a result of adaptation to the colder environment, where body size and the frequency of arrangement 2st have changed in concert. Finally, a heat knockdown test revealed that the lowland population was significantly more resistant to high temperature than the highland one. In conclusion, we suggest that temperature has been an important selective agent causing adaptive differentiation between these two populations. We also suggest that the activity rhythms of the two populations have diverged as a consequence of behavioral evolution, that is, through avoidance of stressful temperatures as a mean of thermal adaptation. PMID- 11392393 TI - Life-history changes that accompany the transition from sexual to parthenogenetic reproduction in Drosophila mercatorum. AB - In spite of the predicted genetic and ecological costs of sex, most natural populations maintain sexual reproduction, even those capable of facultative parthenogenesis. Unfertilized eggs from natural populations of Drosophila mercatorum occasionally develop into viable adults, but obligately parthenogenetic populations are unknown in this species. To evaluate the microevolutionary forces that both favor and constrain the evolution of parthenogenesis in D. mercatorum, we have measured parthenogenetic rates across a natural, sexually reproducing population and characterized the life-history changes that accompany the transition from sexual to parthenogenetic reproduction in laboratory strains. A highly significant difference in parthenogenetic rate was found between two populations in close geographic proximity, with increased rate found with lower population density. Laboratory strains of parthenogenetic females suffered increased mortality and reduced egg viability relative to their virgin counterparts from a sexual strain. Lifetime egg production was similar across all strains, but a shift in peak egg production to an earlier age also occurred. The combination of these life-history traits resulted in a higher net reproductive value for sexual females, but because they also had a longer generation time, intrinsic rate of increase was not as dramatically different from parthenogenetic females. In environments with high early mortality, there may be no fitness disadvantage to parthenogenesis, but the predicted ecological advantage of a twofold increase in intrinsic rate of increase was not realized. These results support the theory of Stalker (1956) that parthenogenesis is favored in environments in which sexual reproduction is difficult or impossible. PMID- 11392394 TI - Unpredictable offspring survivorship in the damselfly, Megaloprepus coerulatus, shapes parental behavior, constrains sexual selection, and challenges traditional fitness estimates. AB - Evolutionary biologists typically assume that the number of eggs fertilized or developing embryos produced is correlated with an individual's fitness. Using microsatellite markers, we document for the first time estimates of realized fitness quantified as the number of offspring surviving to adulthood in an insect under field conditions. In a territorial damselfly whose males defend tree hole oviposition sites, patterns of offspring survivorship could not be anticipated by adults. Fewer than half of the parents contributing eggs to a larval habitat realized any reproductive success from their investment. The best fitness correlate was the span over which eggs in a clutch hatched. Among parents, female fecundity and male fertilization success were poor predictors of realized fitness. Although body size was correlated with female clutch size and male mating success, larger parents did not realize greater fitness than smaller ones. The uncoupling of traditional fitness surrogates from realized fitness provides strong empirical evidence that selection at the larval stage constrains selection on mated adults. PMID- 11392395 TI - Genetics, experience, and host-plant preference in Eurosta solidaginis: implications for host shifts and speciation. AB - Host-associated mating is crucial in maintaining the partial reproductive isolation between the host races of Eurosta solidaginis (Diptera: Tephritidae), a fly that forms galls on Solidago altissima and S. gigantea. (We refer to flies reared from S. gigantea as gigantea flies and those reared from S. altissima as altissima flies.) We measured the host preference of males and females of both host races, F1 hybrids between the host races, F2, and backcrosses to both host races. Male and female altissima flies and female gigantea flies had high host fidelity, whereas male gigantea flies had low host fidelity. This result suggests that there may be gene flow between the host races due to nonassortative mating that occurs when male gigantea mate with altissima females on S. altissima. This indicates assortative-mating mechanisms in addition to host-associated mating are required to produce the partial reproductive isolation between the host races that has been observed. Nongenetic factors had no influence on host preference. Larval conditioning did not influence host preference: reciprocal F1 hybrids reared in S. altissima and S. gigantea both preferred S. gigantea. Adult experience had no impact on host preference: females preferred their natal host plant regardless of which host they encountered first as an adult. The hypothesis that maternal effects influence preferences was rejected because male and female flies did not show a consistent preference for the host plant of their mother. We also found no evidence that preference was a sex-linked trait because F1 and backcrosses to the host races with different combinations of X chromosomes from the two host races preferred S. gigantea. Our results indicate that host preference is not determined by a large number of genes because preference of hybrids did not correspond to the proportion of the genome derived from each host race. The strength of the ovipuncture preference for S. gigantea by gigantea females, the females of both reciprocal F1 hybrids, the backcross to gigantea, and F2s indicates that preference is inherited nonadditively at a limited number of loci. The F1 female hybrids, however, had a weaker host preference for S. gigantea than the pure gigantea host race, indicating that there may be incomplete dominance or modifier loci. Males had different host preference patterns than females, with individual male gigantea and male F1 hybrids usually exhibiting preference exclusively for S. gigantea or S. altissima. One hypothesis explaining the difference in host preference between males and females is that the same gene influences both female and male host preference, but it is a sex influenced gene. Thus, males carrying the gene for S. gigantea preference have an intermediate host preference, whereas females have a strong host preference to S. gigantea. In summary, we found that the host preference that produces host associated mating is inherited nonadditively at a relatively small number of loci on autosomal genes. This mode of inheritance meets the assumptions of models of sympatric speciation, indicating that the host races could have evolved in sympatry. PMID- 11392396 TI - Evolutionary dynamics of host-plant specialization: a case study of the tribe Nymphalini. AB - Two general patterns that have emerged from the intense studies on insect-host plant associations are a predominance of specialists over generalists and a taxonomic conservatism in host-plant use. In most insect-host plant systems, explanations for these patterns must be based on biases in the processes of host colonizations, host shifts, and specialization, rather than cospeciation. In the present paper, we investigate changes in host range in the nymphalid butterfly tribe Nymphalini, using parsimony optimizations of host-plant data on the butterfly phylogeny. In addition, we performed larval establishment tests to search for larval capacity to feed and survive on plants that have been lost from the female egg-laying repertoire. Optimizations suggested an ancestral association with Urticaceae, and most of the tested species showed a capacity to feed on Urtica dioica regardless of actual host-plant use. In addition, there was a bias among the successful establishments on nonhosts toward plants that are used as hosts by other species in the Nymphalini. An increased likelihood of colonizing ancestral or related plants could also provide an alternative explanation for the observed pattern that some plant families appear to have been colonized independently several times in the tribe. We also show that there is no directionality in host range evolution toward increased specialization, that is, specialization is not a dead end. Instead, changes in host range show a very dynamic pattern. PMID- 11392397 TI - Physiological bases of genetic differences in cannibalism behavior of the confused flour beetle Tribolium confusum. AB - Physiological causes of genetic differences in cannibalism were examined to gain a better understanding of constraints on behavior evolution. Cannibalism has complex population level consequences in Tribolium confusum, including dramatic effects on population size. Laboratory strains with low and high cannibalism rates, obtained through inbreeding, have maintained distinct levels of cannibalism for over two decades even in the absence of artificial selection to maintain the differences. Why strains differ in their cannibalism rates was examined by measuring: (1) the nutritional benefit from cannibalism in both nutritionally good and poor environments, and (2) the possibility that eggs are an important source of water. How strains achieve differences in cannibalism was examined by testing for differences between strains in their ability to find eggs and in their tendency to eat eggs. Beetles from both strains survive equally well in a nutritionally good environment, but they accomplish this in different ways. The low cannibalism strain has high survivorship with and without cannibalism. The high cannibalism strain has low survivorship when not fed eggs and survivorship equivalent to the low cannibalism strain when fed eggs, suggesting it compensates for poor nutritional adaptation by eating eggs. The strains also differ in feeding behavior; beetles from the high cannibalism strain have a higher appetite for eggs. Beetles from the two strains did not differ in locomotor activity, search efficiency, or need for water. The observed behavioral and nutritional differences may contribute to the maintenance of different levels of cannibalism. PMID- 11392398 TI - The evolutionary enigma of bonefishes (Albula spp.): cryptic species and ancient separations in a globally distributed shorefish. AB - Many examples of cryptic marine species have been demonstrated with biochemical and molecular studies. In most cases, a broadly distributed taxon is actually a group of sibling species that can be distinguished (upon closer examination) by ecological or morphological characters. Fishes of the family Albulidae constitute a notable exception. Bonefish (Albula spp.) morphology and ecology are highly conserved around the globe, and their extended pelagic larval stage could allow population connections on a vast geographic scale. Based on this perceived homogeneity, bonefishes were classified as a single pantropical species, A. vulpes. However, allozyme studies of Hawaiian populations indicated that two sympatric species (A. glossodonta and A. neoguinaica) are included in the synonymy of A. vulpes. To ascertain the number and distribution of evolutionary partitions in Albula, we surveyed 564 bp of mitochondrial DNA (mtDNA) cytochrome b from 174 individuals collected at 26 locations. Sequence comparisons reveal eight deep lineages (d = 5.56-30.6%) and significant population structure within three of the four lineages that could be tested (phiST = 0.047-0.678). These findings confirm the genetic distinctiveness of the three species noted above and invoke the possibility of five additional species. Clock estimates for mtDNA indicate that these putative species arose 4-20 million years ago. Distinct evolutionary lineages coexist in several sample locations, yet show little morphological or ecological differentiation in sympatry. Thus, bonefish species seem to defy the evolutionary conventions of morphological differentiation over time and ecological displacement in sympatry. Despite multiple cases of sympatry, sister-taxa relationships inferred from mtDNA indicate that divergence in allopatry has been the predominant speciation mechanism in Albula. Stabilizing selection in the homogeneous habitat occupied by bonefishes (tropical sand flats) could promote the retention of highly conserved morphology and ecology. PMID- 11392399 TI - Delayed costs of an induced defense in tadpoles? Morphology, hopping, and development rate at metamorphosis. AB - Models for the evolution of plasticity predict that individuals having phenotypes induced by exposure to enemies should experience relatively low fitness when enemies are absent. However, costs of induced phenotypes have been difficult to find in both plants and animals, perhaps because costs are expressed at later stages in the life cycle. We searched for delayed costs of an induced defense in larvae of the water frog Rana ridibunda, which exhibits strong phenotypic responses to predators. Tadpoles grew to metamorphosis in outdoor artificial ponds, in either the presence or absence of Aeshna dragonfly larvae confined within cages. We collected metamorphs at forelimb emergence, estimated their development rate until tail resorption was complete, and measured their body and leg shape and hopping performance. Development rate through metamorphosis reflects the duration of a transitional period during which metamorphs are especially vulnerable to predators, and hopping performance may reflect ability to escape predators. Froglets from the dragonfly treatment lost mass through metamorphosis significantly faster than those from predator-free ponds, but they resorbed their tails at about the same rate, despite the fact that their tails were relatively large to begin with. Froglets developing from predator-induced tadpoles had shorter, more muscular legs, and hopped 5% longer distances (difference not significant). Therefore, producing an induced defense against insect predators during the tadpole stage did not exact a cost during or immediately after metamorphosis; if anything, tadpoles with the predator-induced phenotype gave rise to more vigorous froglets. These results focus attention on other costs of the induced phenotype, as well as alternative explanations for plasticity that do not rely on direct fitness trade-offs. PMID- 11392400 TI - Reproductive asynchrony increases with environmental disturbance. AB - While it is widely recognized that the manner in which organisms adjust their timing of reproduction reflects evolutionary strategies aimed at minimizing offspring mortality or maximizing reproductive output, the conditions under which the evolutionarily stable strategy involves synchronous or asynchronous reproduction is a matter of considerable discord. A recent theoretical model predicts that whether a population displays reproductive synchrony or asynchrony will depend on the relative scales of intrinsic regulation and environmental disturbance experienced by reproducing individuals. This model predicts that, under conditions of negligible competition and large-scale environmental perturbation, evolution of a single mixed strategy will result in asynchronous reproduction. We tested this prediction using empirical data on large-scale climatic fluctuation and the annual timing of reproduction by three species of flowering plants covering 1300-population-years and four degrees of latitude in Norway. In agreement with model predictions, within populations of all three species reproductive asynchrony increased with the magnitude of large-scale climatic perturbation, but bore no relation to the strength of local density dependence. These results suggest that mixed evolutionarily stable strategies can arise from the interplay of combinations of agents of selection and the scale at which they operate; hence it is fruitless to associate synchronous versus asynchronous timing with particular single factors like climate, competition, or predation. PMID- 11392401 TI - Male genotype affects female longevity in Drosophila melanogaster. AB - Several recent studies suggest that interactions with conspecific males can reduce the longevity of female Drosophila melanogaster or support the idea that male and female fitness components are involved in antagonistic interactions. Here we report that males from third-chromosome isogenic lines demonstrated significant genetic variation in male reproductive performance and in the longevity of their mates. Increased male performance was marginally significantly associated with one measure of increased female survival rate. However, there was no indication of tradeoffs or negative correlations between male reproductive success and female survival. We discuss alternative hypotheses for the cause of the induced variation in female longevity. PMID- 11392402 TI - Adult female Drosophila pseudoobscura survive and carry fertile sperm through long periods in the cold: populations are unlikely to suffer substantial bottlenecks in overwintering. AB - To assess whether, while overwintering, natural populations of Drosophila pseudoobscura are likely to experience substantial bottlenecks in their numbers and genotypes, laboratory tests of the cold sensitivities of each stage of the life history and reproduction were undertaken. Three genetically distinctive lineages established from flies caught at high elevation were used for testing in temperatures likely to persist in protected pockets of fermenting deciduous leaf fall in overwintering sites. Sensitivities to cold of each stage in development were measured as frequencies of survival to adulthood following a period in 5 degrees C in a particular stage. The cold sensitivity of adults was measured as the survival in and following cold stays in adulthood. It was found that cold sensitivity decreases as development progresses, but that only adults (females more than males) are able to withstand long periods in the cold. The cold sensitivity of reproductive capacity of males was scored as their success in mating following a two-month cold stay, and of females as the numbers laying fertile eggs following periods of months in the cold. Both males and females maintain reproductive capacity. Of particular significance, however, is that even after six months in the cold females are able to restart production of eggs and these eggs may be fertilized by the sperm of matings prior to their cold stay. Thus, a substantial proportion of overwintering genomes must be those of adult females and those of the sperm carried by females from matings in the previous summer. This simple finding strongly suggests that populations are not likely to suffer substantial bottlenecks while overwintering. Further, it indicates how arrays of genetic variation may be maintained through winters and largely avoid winter selective pressures. Frequent migration between populations is therefore not required to maintain the variation commonly found in populations throughout the species range. PMID- 11392403 TI - Considerations and implications of LASIK in children. PMID- 11392404 TI - More on steroids after strabismus surgery. PMID- 11392405 TI - Critical age of botulinum toxin treatment in essential infantile esotropia. AB - PURPOSE: To assess the results of botulinum toxin treatment in 60 consecutive children with essential infantile esotropia. METHODS: Bilateral simultaneous injection of botulinum toxin into the medial rectus muscle was performed in 60 patients under direct visualization with an "open sky" technique. Fluothane/sevoflurane insufflation anesthesia was used. Each patient underwent a single bilateral botulinum toxin injection. Patient age at the time of injection ranged from 5-8 months. RESULTS: Mean patient age at the time of treatment for the 88% of patients who gained a good alignment (within +10 prism diopters [delta] of residual esotropia) was 6.5 months, while mean patient age at time of injection for the 12% of patients who were undercorrected or the deviation relapsed was 7.8 months. Follow-up averaged 5.2 years (range: 2-9 years, SD 2). No variation of the angle of strabismus was observed after 6 months from injection. In some patients with hyperopic refraction, plus lens corrections were prescribed during follow-up to stabilize the alignment. CONCLUSION: Botulinum toxin can be effective in essential infantile esotropia when children are treated by age 7 months. PMID- 11392406 TI - Development of two stable oral suspensions of levodopa-carbidopa for children with amblyopia. AB - PURPOSE: To develop stable liquid dosage forms of levodopa-carbidopa for use in children with amblyopia. METHODS: Levodopa (100 mg) and carbidopa (25 mg) tablets were used to prepare the suspensions at 5 and 1.25 mg/mL, respectively. For each suspension, five bottles were stored at 25 degrees C and five at 4 degrees C. Three samples were taken from each bottle at 0, 7, 14, 28, 42, 56, 70, and 91 days (n=15). Levodopa-carbidopa concentrations for each sample were measured in duplicate by validated and stability-indicating high-performance liquid chromatographic method. RESULTS: The mean concentrations of levodopa-carbidopa in Ora Plus/Ora Sweet suspensions were 96% and 92% of the initial concentrations for 28 days at 25 degrees C, and 94% and 93% for 42 days at 4 degrees C. In the suspension containing ascorbic acid, the mean concentrations of levodopa carbidopa were above 93% and 92% for 14 days at 25 degrees C, and 93% and 92% for 28 days at 4 degrees C. This liquid formulation was administered to 15 children (mean age: 5.6+/-1.4 years) with amblyopia. The number and type of adverse effects were similar in the levo-dopa-carbidopa versus placebo group. CONCLUSION: Levodopa-carbidopa in extemporaneous suspensions prepared in Ora Plus and Ora Sweet were stable for 28 days when stored at 25 degrees C and for 42 days at 4 degrees C. Our data suggest our liquid formulation may be used safely in children with amblyopia. PMID- 11392407 TI - Complications of ocular paintball injuries in children. AB - PURPOSE: To evaluate the ocular complications of paintball injuries in children. METHODS: The clinical course of four children with traumatic ocular paintball injuries was evaluated. All patients underwent a complete ocular examination. Their age, injuries sustained, surgical procedure(s) performed, presence of protective eyewear at the time of injury, and final visual outcome was assessed. The presence of directly related anterior and posterior segment abnormalities were also evaluated. RESULTS: Four boys sustained traumatic paintball injuries. Average patient age was 11.25 years (range: 10-12 years). None of the children were wearing ocular or facial protection at the time of the initial injury. All patients had hyphema and traumatic cataract, and some form of retinal pathology (vitreous hemorrhage, epiretinal membrane, retinal hemorrhage, and choroidal rupture). One child had a partial-thickness corneal laceration that did not require surgical intervention. All other patients underwent ophthalmic surgery. Final visual acuity was 20/30 or better in two patients, and 20/100 or worse in the others. The cause of decreased visual acuity in these children was directly related to macular pathology. CONCLUSION: Ocular injuries resulting from paintball impact are often severe and usually occur when the participants are not wearing eye protection or this protection becomes dislodged. Treatment of these injuries is sometimes limited to an attempt to salvage what remains of useful vision. Unfortunately, most of these sports-related injuries could have been prevented if patients wore adequate eye protection when involved in this sport. PMID- 11392408 TI - Tolerance of patients and postoperative results: topical anesthesia for strabismus surgery. AB - PURPOSE: To assess patient tolerance and the postoperative results of intraoperative adjustment of strabismus surgery under topical anesthesia. METHODS: Twenty-four patients with strabismus underwent rectus muscle recession under topical anesthesia without the use of additional systemic analgesics and sedatives. Patient tolerance was estimated according to the sites of procedure. In addition, the angle of strabismus was measured 1 day, 6 weeks, and 1 year postoperatively. RESULTS: Of the 24 patients identified (11 men and 13 women; mean age: 24 years), 16 had exotropia, 7 were esotropic, and 1 was hypertropic. During the operation, 17 patients never expressed any pain, and 7 patients complained of discomfort on muscle manipulation. Postoperatively, the success rate was 95.8% (23 patients) the first day after surgery, 79.2% (19 patients) after 6 weeks, and 70.8% (17 patients) after 1 year. CONCLUSION: Intraoperative adjustment performed with the patient under topical anesthesia is recommended in all cooperative cases of strabismus surgery and is comfortable for patients. PMID- 11392409 TI - Astigmatism in infancy and childhood. AB - PURPOSE: To evaluate the prevalence of astigmatism in infancy and childhood in a Spanish population. METHODS: A total of 478 children with astigmatism from Valencia Province, Spain were examined. The patients, ranging in age from 2-12 years, were classified into groups according to age. Astigmatism error and axis orientation were determined using noncyclopegic retinoscopy over each eye. RESULTS: The overall prevalence of astigmatism (> or = 1.00 diopter of cylinder) decreased in relation to increasing age from 44.3% to 5.2%. The prevalence of against-the-rule astigmatism decreased from 61.5% (at 2 years) to 33.4% (at 12 years), with-the-rule astigmatism increased in prevalence from 23.1% (at 2 years) to 66.6% (at 12 years), and oblique astigmatism decreased with age from 15.4% (at 2 years) and 0% (at 12 years). CONCLUSION: These results do not vary from those observed in other populations. Factors such as race, nutrition, and environmental condition did not vary from the literature regarding the prevalence of astigmatism in infants and children. PMID- 11392410 TI - Potential use of LASIK in children. PMID- 11392411 TI - Slipped medial rectus muscle secondary to orbital hemorrhage following strabismus surgery. PMID- 11392412 TI - Intermittent esotropia as equivalent of absence in epilepsy. PMID- 11392413 TI - Amelanotic malignant choroidal melanoma in a 10-year-old girl. PMID- 11392414 TI - Aniridia and Down syndrome. PMID- 11392415 TI - Spontaneous expulsion of a subconjunctival cysticercus cyst. PMID- 11392416 TI - Pancreatic ultrasonography. AB - Pancreatic abnormalities usually are detected on US when it is used for screening patients with abdominal pain and for assessment of the gallbladder and bile ducts. Pancreatic visualization is limited by bowel gas, but with experienced sonographers and newer techniques, including harmonic imaging and oral contrast US, diagnosis of pancreatic abnormalities has significantly improved compared with earlier reports. Appropriate initial diagnosis by US can tailor further investigation, and US-guided biopsy may establish definitive diagnosis. PMID- 11392417 TI - Computed tomography for pancreatic diseases. AB - CT has evolved over 24 years as an effective cross-sectional imaging modality for benign and malignant pancreatic diseases. Multiphase helical scanners with rapid infusion of intravenous contrast medium allow for the delineation of pancreatic parenchymal, arterial, and venous studies with identification of malignant and benign lesions. Therapeutic applications abound, and CT drainage of postoperative fluid collections, pseudocysts, and abscesses will have an increasing role in the new millennium. PMID- 11392418 TI - MR imaging of the pancreas. AB - Current MR imaging technology offers the surgeon diagnostic information about pancreatic diseases. This article reviews the basics of MR imaging formation and the rationale for the different types of imaging sequences that comprise a comprehensive pancreaticobiliary examination. Clinical examples include evaluation of pancreatic neoplasms, acute and chronic pancreatitis, and congenital abnormalities. PMID- 11392419 TI - Endoscopic retrograde cholangiopancreatography and the pancreas: when and why? AB - Historically, endoscopic retrograde cholangeopancreatography (ERCP) has played a unique role as a diagnostic and therapeutic modality for diseases of the pancreas. Despite the advent of new imaging techniques, ERCP continues to play a central role in the evaluation and treatment of pancreatitis of various causes. Endoscopic cholangiography and stenting remain mainstays in the palliation of inoperable obstructive jaundice in patients with cancer of the pancreas. PMID- 11392420 TI - Endoscopic ultrasonography in benign pancreatic disease. AB - It seems that EUS will most likely become the gold standard technique to diagnose chronic pancreatitis not diagnosed on plain radiography, standard transcutaneous sonography, or CT scanning. Because of its low risk and increased sensitivity, it will replace ERCP as a diagnostic test for this condition. Confirmation with cytology may be beneficial for indeterminate cases. EUS will also have an important role in determining the management of cystic lesions in the pancreas. EUS seems to be very effective in determining which cystic lesions have malignant potential. If a cyst appears malignant or produces symptoms, it requires resection and therefore does not require FNA. Prediction of the clinical course for cysts of indeterminate nature requires EUS-guided FNA and analysis of fluid. EUS, although quite accurate in diagnosing CBD stones, has a more limited role in diagnosis and management of stone disease because of current limitations of therapeutic maneuvers, which can be performed at the same time. In general, patients with probable CBD stones or sludge require therapeutic ERCP. Therefore, ERCP is the preferred initial test to diagnose and simultaneously treat these disorders. Patients with a low suspicion for CBD stones, or patients with relative contraindications to ERCP (i.e., pregnancy or bleeding disorders), can be evaluated first with EUS to determine whether further invasive treatment is required. EUS seems to be complementary to therapeutic ERCP for the aspiration and drainage of cysts and pseudocysts. Although celiac plexus nerve blocks using EUS-guided injection of neurolytic agents seems to be more effective than other nerve block techniques, surgical bypass or resection is likely to continue as the primary method of treatment of patients with pain from chronic pancreatitis or those who do not respond to endoscopic stenting when there is a dominant stricture that can be bypassed. PMID- 11392421 TI - The role of nuclear medicine in the evaluation of pancreatic disease. AB - F-18 FDG PET in patients with nonendocrine pancreatic cancer and somatostatin receptor imaging in patients with endocrine pancreatic cancer have an important role in detecting or confirming the presence of a mass in the pancreas--a crucial step in the management of these patients. Both agents also have an important role in staging and in defining which patients are good candidates for resection surgery. Somatostatin receptor imaging also has a crucial role in selecting from the various systemic therapeutic options available. I-131 MIBG therapy can be of value therapeutically, mostly palliative, in patients who demonstrate a markedly elevated concentration of tumor radioactivity. PMID- 11392422 TI - Angiography and interventional radiology: percutaneous approaches to benign pancreatic disorders. AB - Interventional radiology contributes several avenues for the diagnosis and treatment of benign pancreatic disorders. Biopsy can provide supporting evidence for benign or equivocal lesions. Aspiration of collections suspected of infection can yield specific bacteriologic diagnosis. Bleeding complications of pancreatitis can be treated directly by transcatheter embolization. PMID- 11392423 TI - Diagnostic and interventional laparoscopy and intraoperative ultrasonography in the management of pancreatic disease. AB - The laparoscopic management of pancreatic disorders has evolved dramatically from its inception in 1911 and its rediscovery in the 1970s. Although investigators once proclaimed that "it seems unlikely that laparoscopy will have any more than an extremely limited use in the investigation of pancreatic disorders," laparoscopy and LUS now have a well-recognized role in the staging of pancreatic cancer and an increasing part in the management of benign pancreatic disease at many institutions. Although the appropriate role of LS and LUS is debatable, the development and refinement of laparoscopic techniques and instrumentation and the improvement of noninvasive diagnostic modalities will provide new data, increase the rate of resection at laparotomy, and allow surgeons to treat a broader range of pancreatic disease by minimally invasive methods. The value of LS and LUS for benign and malignant pancreatic disorders has been clearly demonstrated, but the inevitable issues of hospital resource, operative expertise, and surgical philosophy will ultimately determine the role of laparoscopy and LUS in clinical practice. PMID- 11392424 TI - Pathogenesis of alcohol-induced chronic pancreatitis: facts, perceptions, and misperceptions. AB - The pathogenesis of chronic pancreatitis secondary to chronic alcoholism is not fully understood. A major hurdle in the understanding of the pathogenesis is the inability to study early lesions of the pancreas and the sequential changes. Facts are few; observations are many. Each new hypothesis argues against all previous hypotheses; however, clinical chronic pancreatitis is initiated by one or more of the mechanisms. Good experimental models for alcoholic pancreatitis are not available, limiting the ability to study the pathogenesis. Additional studies on genetic markers and immunologic mechanisms might explain acinar cell injury, which seems to be the earliest lesion in most, if not all, types of chronic pancreatitis. Opie's common channel and obstruction regurgitation theories seem unrelated to chronic pancreatitis. Although biochemical changes of the pancreatic secretion in alcoholic patients promote protein-plug formation, evidence is too weak to consider protein plug as the earliest change. The theory of necrosis of the acinar cell by some unknown mechanism, subsequently leading to fibrosis, is gaining support; however, it is clear that the pathogenesis of alcoholic pancreatitis is not yet fully understood. PMID- 11392425 TI - An overview of pancreatic pseudocysts: the emperor's new clothes revisited. AB - The diminished role of surgery and increased role of nonoperative interventional therapy for pancreatic pseudocysts is discussed. The natural history supports prolonged observation for most asymptomatic pseudocysts. PMID- 11392426 TI - Pancreatic pseudocysts and fluid collections: percutaneous approaches. AB - Radiological imaging and intervention play important roles in the management of pancreatic fluid collections and pseudocysts. Computed tomography evaluation of the severity of pancreatitis and assessment of its course are now routine. Percutaneous drainage of pancreatic pseudocysts and abscesses is commonly performed as an adjunct to surgical treatment and is frequently definitive therapy. Percutaneous debridement of pancreatic necrosis has recently emerged as a viable alternative to open surgical treatment. PMID- 11392427 TI - Endoscopic management of pancreatic pseudocysts. AB - This article reviews the history of and the authors' experience with endoscopic management of pancreatic pseudocysts. Discussion includes pseudocyst enterostomy and results, the transpapillary method, complications, endoscopic versus surgical and percutaneous therapy, and drainage of infected pseudocysts and pancreatic necrosis. PMID- 11392428 TI - Surgical treatment of pancreatic pseudocysts. AB - The surgical treatment of pancreatic pseudocysts (PC) has a less important role now that endoscopic and radiologic intervention can effectively treat PC. Surgery may be needed to correct the underlying cause--usually an obstructed or disrupted pancreatic duct. PMID- 11392429 TI - Endoscopic therapy for chronic pancreatitis. AB - Endoscopic therapy for chronic pancreatitis is feasible and effective in selected patients. The management of pain and ductal obstruction is most effective if reversal of the obstructive process--stricture or stone--is successful and durable. Multiple endoscopic modalities are available, and new technologies will continue to advance the capabilities of therapeutic pancreatic endoscopists. Adjunctive treatments, such as ESWL, enhance the success of these techniques. These varied therapies, although attractive and theoretically sensible, have not been compared in a randomized, controlled fashion with standard surgical therapies. In this sense, they remain experimental. Nonetheless, these techniques are widely applied in advanced endoscopy centers worldwide, and uncontrolled individual series are expected to continue to expound on and demonstrate the effectiveness of these minimally invasive interventions until randomized, prospective studies become available. PMID- 11392430 TI - Surgery and chronic pancreatitis. AB - It is hoped that, in this millennium, chronic pancreatitis will be diagnosed earlier in the course of the disease process. Improved axial imaging of the pancreatic duct and pancreatic parenchyma will diminish the need for other invasive tests. Surgical procedures are directed at pancreatic duct decompression or resection of the pancreas (head, body or tail) or, infrequently, total pancreatectomy. Pain relief in 75% to 90% is the general rule, with diabetes developing subsequently in as many as 33% of patients. Surgery for chronic pancreatitis is effective in correcting sequelae of pancreatic fibrosis. Endoscopic stenting of the pancreatic and bile duct is used more frequently today. Until their place is ascertained, careful performance of surgery will continue to be a mainstay of treatment. PMID- 11392431 TI - Surgical management of chronic pancreatitis at the Mayo Clinic. AB - The authors' approach to the overall surgical management of chronic pancreatitis is to treat complications, that is, pain and, less commonly, obstruction and bleeding. The authors' practice is to exhaust nearly all forms of nonsurgical intervention before suggesting a surgical approach. Nonresponders are then evaluated for severity of pain, interference of quality of life, and presence of chemical dependency. Appropriate candidates undergo imaging examinations to determine the primary site of disease, presence of pancreatic ductal dilatation, and associated peripancreatic complications. The surgical treatment approach involves classic lines of proximal resection (pylorus-preserving pancreaticoduodenectomy) for small duct disease and lateral pancreaticojejunal drainage for a dilated pancreatic duct. The authors have not yet routinely adopted the duodenum-preserving head resections of Beger and Frey, or thoracoscopic transthoracic splanchnicectomy, but they remain open-minded and avidly await good confirmatory, independent trials of these promising surgical interventions. PMID- 11392432 TI - Pancreas divisum: endoscopic therapy. AB - Pancreas divisum is a common congenital variation that can be associated with pancreatic disease. Symptomatic patients with divisum must be classified according to clinical presentation and morphologic findings. Response to endoscopic therapy is best in patients with ARP, of whom 75% benefit. Results in patients with chronic pancreatitis and pain but without objective pancreatitis are mixed, and patients should be carefully selected. PMID- 11392433 TI - Surgery and pancreas divisum: a dwindling role? AB - Pancreas divisum has generated varying enthusiasm regarding operative intervention. Applying similar principles to divisum surgery as for the surgical treatment of chronic pancreatitis will yield a better outcome than using subjective symptoms. PMID- 11392434 TI - Effect of stimulus intensity level on auditory middle latency response brain maps in human adults. AB - Auditory middle latency response (AMLR) brain maps were obtained in 11 young adults with normal hearing. AMLR waveforms were elicited with monaural clicks presented at three stimulus intensity levels (50, 70, and 90 dB nHL). Recordings were made for right and left ear stimulus presentations. All recordings were obtained in an eyes open/awake status for each subject. Peak-to-peak amplitudes and absolute latencies of the AMLR Pa and Pb waveforms were measured at the Cz electrode site. Pa and Pb waveforms were present 100 percent of the time in response to the 90 dB nHL presentation. The prevalence of Pa and Pb to the 70 dB nHL presentation varied from 86 to 95 percent. The prevalence of Pa and Pb to the 50 dB nHL stimulus never reached 100 percent, ranging in prevalence from 77 to 68 percent. No significant ear effect was seen for amplitude or latency measures of Pa or Pb. AMLR brain maps of the voltage field distributions of Pa and Pb waveforms showed different topographic features. Scalp topography of the Pa waveform was altered by a reduction in stimulus intensity level. At 90 dB nHL, the Pa brain map showed a large positivity midline over the frontal and central scalp areas. At lower stimulus intensity levels, frontal positivity was reduced, and scalp negativity over occipital regions was increased. Pb scalp topography was also altered by a reduction in stimulus intensity level. Varying the stimulus intensity significantly altered Pa and Pb distributions of amplitude and latency measures. Pa and Pb distributions were skewed regardless of stimulus intensity. PMID- 11392435 TI - Effects of lexical factors on word recognition among normal-hearing and hearing impaired listeners. AB - An investigation was conducted to examine the effects of lexical difficulty on spoken word recognition among young normal-hearing and middle-aged and older listeners with hearing loss. Two word lists, based on the lexical characteristics of word frequency and neighborhood density and frequency (Neighborhood Activation Model [NAM]), were developed: (1) lexically "easy" words with high word frequency and a low number and frequency of words phonemically similar to the target word and (2) lexically "hard" words with low word frequency and a high number and frequency of words phonemically similar to the target word. Simple and transformed up-down adaptive strategies were used to estimate performance levels at several locations on the performance-intensity functions of the words. The results verified predictions of the NAM and showed that easy words produced more favorable performance levels than hard words at an equal intelligibility. Although the slopes of the performance-intensity function for the hearing impaired listeners were less steep than those of normal-hearing listeners, the effects of lexical difficulty on performance were similar for both groups. PMID- 11392436 TI - Reliability of the mismatch negativity in the responses of individual listeners. AB - The mismatch negativity (MMN) was recorded from 12 normal adults during four biweekly sessions. Responses were elicited by a synthetically generated speech contrast (/dalpha/-/galpha/) that all listeners discriminated with at least 90 percent accuracy. Standard and deviant waveforms were replicable across sessions for all listeners; however, replicability of the derived difference waveforms was poor. Of greater importance, the MMN identification rate was too low (29%) to allow reliability to be evaluated. The implications that these findings may have on clinical applicability are discussed. PMID- 11392437 TI - Performance on a Spanish picture-identification task using a multimedia format. AB - An option for estimating the word-recognition performance of patients who do not speak English as a first language involves using auditory materials, presented in the patient's native language, in conjunction with a closed-set response mode incorporating pictures or written words. The advantage of this auditory/visual paradigm is that the audiologist is not required to know the foreign language and is therefore not required to judge the accuracy of an oral response to speech stimuli in a foreign language. Spanish auditory/visual materials, known as the Spanish Picture-Identification Task, were developed to be used in a computer driven multimedia administration and scoring format. Performance data, both in open- (word-recognition) and closed-set (word-identification) response modes, were established for the Spanish Picture-Identification Task using subjects whose first language was Spanish. The results from the open-set paradigm indicate that the Spanish Picture-Identification Task word lists are essentially equivalent to conventional Spanish and English materials used for word recognition. Findings from the closed-set conditions indicate that the Spanish Picture-Identification Task materials are appropriate for estimating the word-identification abilities of Spanish-speaking adult listeners. PMID- 11392438 TI - Listeners who prefer monaural to binaural hearing aids. AB - Four patients who preferred monaural as compared with binaural amplification were evaluated. For these patients, audiometric data, recognition performance on a dichotic digit task, and monaural and binaural hearing aid performance using four amplification strategies (National Acoustic Laboratories-Revised, a speech in noise algorithm, multiple-microphone arrays, and frequency modulated [FM]) are described. The results of dichotic testing using a one-, two-, and three-pair dichotic digit task in free- and directed-recall conditions indicated a left-ear deficit for all subjects that could not be explained by peripheral auditory findings or by a cognitive-based deficit. The results of soundfield testing using a speech in multitalker babble paradigm indicated that when listening in noise, there was little difference between aided and unaided word-recognition performance, suggesting that the binaural hearing aids originally fit for each patient were not providing substantial benefit when listening in a competing babble background. Word-recognition performance when aided monaurally in the right ear was superior to performance when aided monaurally in the left ear and when aided binaurally. The only successful binaural amplification strategy was the FM system. The results indicate that listeners with an auditory-based deficit in dichotic listening may function better with a monaural hearing aid fitting or with an assistive listening device such as an FM system. The findings also suggest that a test of dichotic listening is an important component in the evaluation of patients being considered for amplification. PMID- 11392439 TI - Pneumococcal disease in the elderly: what is preventing vaccine efficacy? AB - The effective prevention of Streptococcus pneumoniae infection has a renewed priority in an era in which the emergence of antibacterial-resistant strains has the potential to further compromise efforts to reduce early mortality from invasive pneumococcal infection. Although the 23-valent pneumococcal polysaccharide (PPS) vaccine was approved in the US to prevent respiratory and invasive infection in the elderly and other high-risk populations, the protective efficacy of this vaccine for the growing population of adults aged >65 years remains controversial. The apparent effectiveness of pneumococcal immunisation in clinical studies of elderly adults has varied depending upon whether a reduction in pneumococcal colonisation, pneumonia, bacteraemia or death was used as an outcome. Clinical studies of vaccine efficacy to date suggest that the current pneumococcal vaccine is 56 to 81% effective at preventing invasive pneumococcal infection, and may have additive benefit to influenza vaccine in preventing community-acquired pneumonia, particularly in elderly adults with an increased risk of serious pneumonia requiring hospitalisation. Possible reasons for incomplete protection from pneumococcal infection after immunisation include infection with non-vaccine serotypes, inadequate or ineffective antibody responses, waning of antibody responses, or compromised nonhumoral host defences. Further studies are needed to determine whether: (i) elderly adults who respond poorly to the 23-valent pneumococcal vaccine can be identified prior to immunisation and targeted for study with improved pneumococcal vaccines; (ii) specific nutrient deficiencies can be identified and corrected to improve the immune responsiveness of elderly adults to the PPS vaccine; (iii) newer protein conjugate or DNA pneumococcal vaccines may be more uniformly immunogenic for elderly adults; and (iv) whether smoking cessation reduces the risk of invasive pneumococcal infection in elderly adults. PMID- 11392440 TI - Treatment of systemic fungal infections in older patients: achieving optimal outcomes. AB - Systemic fungal infections are an increasing problem in older adults. For several of the endemic mycoses, this increase is the result of increased travel and leisure activities in areas endemic for these fungi. Immunosuppressive agents, care in an intensive care unit, and invasive devices all contribute to infection with opportunistic fungi. Treatment of systemic fungal infections is usually with an azole or amphotericin B. The preferred regimen depends on the specific fungal infection, the site and the severity of the infection, the state of immunosuppression of the patient and the possible toxicities of each drug for a specific patient. In older adults, drug-drug interactions between the azoles and drugs commonly prescribed for older persons may lead to serious toxicity, and absorption of itraconazole can be problematic. Amphotericin B is associated with significant nephrotoxicity, especially in older adults with pre-existing renal disease, and infusion-related adverse effects. Newer lipid formulations of amphotericin B can obviate some of these toxicities, but their role in the treatment of systemic fungal infections in older adults has not yet been clarified. PMID- 11392441 TI - Optimisation of the management of patients with coronary heart disease and type 2 diabetes mellitus. AB - Type 2 diabetes mellitus is a prevalent disease in Westernised society, and more than 50% of individuals with diabetes mellitus die from cardiovascular causes. The underlying metabolic defect of type 2 diabetes mellitus is a combination of insulin resistance and decreased secretion of insulin by pancreatic beta-cells. Insulin resistance commonly precedes the onset of type 2 diabetes mellitus and is usually associated with a metabolic syndrome including hypertension, dyslipidaemia and obesity. Treatment of known cardiovascular risk factors, including hyperglycaemia, dyslipidaemia, hypertension and smoking, plays a key role in delaying the onset and progression of coronary heart disease (CHD) and other forms of atherosclerosis in patients with diabetes mellitus. Sulphonylureas should be used with caution in patients with CHD but aspirin (acetylsalicylic acid), beta-blockers and ACE inhibitors play an important role in the medical management of patients with established coronary artery disease and diabetes mellitus. Patients with diabetes mellitus represent a higher risk group of patients after both percutaneous and surgical coronary revascularisation and the decision regarding the choice of revascularisation procedure should take into account angiographic characteristics, clinical status and patient preference. Patients presenting with diabetes mellitus and acute myocardial infarction should be considered for reperfusion therapy with either urgent thrombolytic therapy or primary percutaneous coronary intervention. PMID- 11392443 TI - Improved outcomes with antihypertensive medication in the elderly with isolated systolic hypertension. AB - Isolated systolic hypertension affects over 15% of all individuals aged >60 years. In the elderly, systolic hypertension is a major modifiable cardiovascular risk factor. Systolic blood pressure (SBP) is associated with higher risk of an adverse outcome, whereas diastolic blood pressure (DBP) is inversely correlated with total mortality, independent of SBP, highlighting the role of pulse pressure as a risk factor. Three placebo-controlled outcome trials on antihypertensive drug treatment in older patients with isolated systolic hypertension have been published: the Systolic Hypertension in the Elderly Program (SHEP), the Systolic Hypertension in Europe (Syst-Eur) Trial and the Systolic Hypertension in China (Syst-China) Trial. These 3 trials demonstrated the benefit of antihypertensive drug treatment. A meta-analysis was performed by pooling the patients from these 3 trials with a subset of patients with isolated systolic hypertension from 5 other trials in the elderly. The pooled results of 15,693 older patients with isolated systolic hypertension prove that antihypertensive drug treatment isjustified if on repeated clinic measurements SBP is 160 mm Hg or higher. PMID- 11392442 TI - Medical management of patients with refractory carcinoma in situ of the bladder. AB - Bladder cancer is a common genitourinary malignancy and carcinoma in situ (CIS) of the bladder exists as a potentially aggressive variant of the superficial form of the disease. Treatment must reflect the unpredictable nature of this disease entity. In 1976, the use of intravesical Bacillus Calmette-Guerin (BCG) was described for the management of early stage bladder cancer. A subsequent report demonstrated efficacy in a cohort of patients with CIS of the bladder. Since this time, intravesical BCG has been recognised as the initial therapy for CIS of the bladder. Although a 6-week treatment with intravesical BCG has been established as standard therapy in patients with CIS, there has been no consensus as to the subsequent treatment for patients in the setting of failure to initial management with BCG. In addition, a number of reports have demonstrated an increased potential of adverse effects after repeated treatment with intravesical BCG. A variety of alternative immunological and chemotherapeutic agents have been developed in response to the limitations of BCG for patients with refractory CIS of the bladder. At present, valrubicin remains the only agent that is approved by the US Food and Drug Administration for the specific indication of CIS of the bladder unresponsive to intravesical BCG. Although these agents appear promising, the most efficacious therapy remains to be determined. The specific treatment protocol for refractory CIS of the bladder remains elusive. It is ultimately the combined decision of the clinician and patient to determine which course of management is most beneficial. PMID- 11392444 TI - Selective serotonin reuptake inhibitors for late-life depression: a comparative review. AB - Late-life depression is a serious health problem that is challenging to manage but generally responds well to pharmacotherapy. Selective serotonin (5 hydroxytryptamine: 5-HT) reuptake inhibitors (SSRIs), the most commonly prescribed antidepressants, are usually used as first-line agents for elderly patients with depression. Like most drugs, SSRIs have not been widely tested in clinical trials that approximate 'real-life' geriatric situations. However, studies completed to date provide valuable information about the efficacy, safety and tolerability of this class of antidepressants among older patients with depression, including those with depression secondary to stroke or dementia and those with other comorbid physical disorders. Although one SSRI may be more efficacious or better tolerated by elderly patients than another, existing data do not support such claims. However, other distinguishing features may influence the choice of agent. For example, fluoxetine, fluvoxamine and paroxetine are more likely to be involved in significant drug-drug interactions than are citalopram or sertraline. In contrast to the other SSRIs, fluoxetine has a half-life well in excess of 1 day, which can be an advantage when weaning the patient off therapy in that it may reduce the incidence of discontinuation symptoms, but a significant disadvantage if the patient cannot tolerate the drug or experiences an adverse drug-drug interaction. PMID- 11392445 TI - Indobufen: an updated review of its use in the management of atherothrombosis. AB - Indobufen inhibits platelet aggregation by reversibly inhibiting the platelet cyclooxygenase enzyme thereby suppressing thromboxane synthesis. Clinical trials have evaluated the efficacy of oral indobufen in the secondary prevention of thromboembolic complications in patients with or without atrial fibrillation, in the prevention of graft occlusion after coronary artery bypass graft (CABG) surgery and in the treatment of intermittent claudication. In the secondary prevention of thromboembolic events indobufen 200 mg once or twice daily was significantly more effective than no treatment although not as effective as ticlopidine 250 mg once or twice daily, during 1-year nonblind clinical trials. Compared with placebo, indobufen 100 mg twice daily significantly reduced the risk of stroke in a small 28-month trial of patients at increased risk of systemic embolism (50% had atrial fibrillation). Furthermore, in patients with nonrheumatic atrial fibrillation and a recent cerebrovascular event enrolled in the 1-year Studio Italiano Fibrillazione Atriale (SIFA) trial, indobufen 100 or 200 mg twice daily was as effective as warfarin (titrated to produce an international normalised ratio of 2.0 to 3.5) in the secondary prevention of thromboembolic events; the incidences of the composite end-point of major vascular events (10.6 vs 9.0%) and recurrent stroke (5 vs 4%) were similar between treatments. In 2 large 12-month trials, the Studio Indobufene nel Bypass Aortocoronarico (SINBA) and the UK study, indobufen 200 mg twice daily was as effective as aspirin (acetylsalicylic acid) 300 or 325 mg plus dipyridamole 75 mg 3 times daily in the prevention of early and late occlusion of saphenous grafts in patients after CABG surgery. Indobufen 200 mg twice daily for 6 months significantly improved walking capacity compared with placebo, and caused a more pronounced improvement in both pain-free and total walking distance than either pentoxifylline 300 mg or aspirin 500 mg twice daily in separate 6- and 12-month studies of patients with intermittent claudication. Oral indobufen up to 200 mg twice daily was generally well tolerated in >5000 patients with atherosclerotic disease. Adverse events (predominantly gastrointestinal), reported by 3.9% of patients, rarely required withdrawal from treatment. In the SINBA and UK studies, fewer adverse events and less gastrointestinal bleeding were seen with indobufen than with aspirin plus dipyridamole treatment, while in the SIFA trial, noncerebral bleeding events occurred significantly less frequently in indobufen than warfarin recipients (0.6 vs 5.1%) and major bleeding events occurred only in the warfarin group. CONCLUSION: Indobufen is as effective as warfarin in the prophylaxis of thromboembolic events in at risk patients with nonrheumatic atrial fibrillation, as aspirin plus dipyridamole in the prevention of CABG occlusion and may be more effective than aspirin or pentoxifylline in improving walking capacity in patients with intermittent claudication. The improved tolerability profile of indobufen (favourable gastric tolerance and reduced haemorrhagic complications) compared with aspirin 300 to 325 mg 3 times daily or warfarin, in addition to a similar antiplatelet effect, suggests indobufen can be considered a drug with a definite role in the management of atherothrombotic events. In particular, indobufen may be an effective alternative for at risk patients with nonrheumatic atrial fibrillation in whom anticoagulant therapy is contraindicated or who are at higher risk of bleeding. PMID- 11392446 TI - Determinants of paclitaxel uptake, accumulation and retention in solid tumors. AB - This report addresses the determinants of the rate and extent of paclitaxel accumulation in tumors. In a 2-dimensional system such as monolayers where the drug is directly in contact with tumor cells, drug accumulation is determined by the extracellular-to-intracellular concentration gradient, the drug binding to extracellular and intracellular macromolecules, the presence of the mdrl p glycoprotein (Pgp). and the time-dependent and drug concentration-dependent changes in tubulins and cell density. Intracellular pharmacokinetic models were developed to depict the effects of these parameters. Computer simulation results indicate that at the clinically relevant concentration range of 1 to 1,000 nM, (a) the binding affinity and the number of intracellular saturable drug binding sites are important for drug accumulation at low and high extracellular concentrations, respectively, (b) saturation in the drug binding to the high affinity intracellular binding sites (e.g., tubulin/microtubule) occurs at extracellular drug concentration above 100 nM, (c) treatment with 1,000 nM paclitaxel for >4 hr results in increased levels of tubulin/microtubule and consequently increased intracellular drug accumulation, whereas the continued cell proliferation after treatment with low drug concentrations results in reduced intracellular accumulation, and (d) saturation of Pgp in mdr1-transfected cells occurs at the high end of the clinically relevant concentration range. In a 3-dimensional system such as the solid tumor histocultures, which contain tumor cells as well as stromal cells, the drug accumulation into the inner cell layers is determined by the unique properties of solid tumors, including tumor cell density and spatial arrangement of tumor and stromal tissues. Most interestingly, drug penetration is modulated by the drug-induced apoptosis; the reduced cell density due to apoptosis results in an enhancement of the rate of drug penetration into the inner cell layers of solid tumors. In conclusion, the uptake, accumulation, and retention of paclitaxel in solid tumors are determined by (a) factors that are independent of biological changes in tumor cells induced by paclitaxel, i.e., ratio of extracellular and intracellular concentrations, and drug binding to extracellular and intracellular macromolecules, and (b) factors that are dependent on the time- and drug concentration-dependent biological changes induced by paclitaxel, i.e., induction of apoptosis, enhancement of tubulin/microtubule production, and induction of Pgp expression. PMID- 11392448 TI - Progress in the development of alternative pharmaceutical formulations of taxanes. AB - The currently available taxanes paclitaxel (Taxol) and docetaxel (Taxotere) are clinically effective against advanced breast, ovarian and non-small cell lung cancer. Due to their low aqueous solubility, both taxanes posed difficulties to the pharmaceutical scientists with respect to the development of an intravenous dosage form. At present, paclitaxel is formulated in a mixture of 50:50% (v/v) Cremophor EL and dehydrated ethanol. However, this formulation vehicle is associated with a number of pharmacological, pharmacokinetic and pharmaceutical concerns amongst which serious hypersensitivity reactions. This review deals with the attempts made into the development of alternative dosage forms of paclitaxel devoid of the Cremophor EL/ethanol excipients and potential future taxane formulations. PMID- 11392447 TI - Role of formulation vehicles in taxane pharmacology. AB - The non-ionic surfactants Cremophor EL (CrEL) and Tween 80, both used as formulation vehicles of many (anticancer) agents including paclitaxel and docetaxel, are not physiological inert compounds. We describe their biological properties, especially the toxic side effects, and their pharmacological properties, such as modulation of P-glycoprotein activity. In detail, we discuss their influence on the disposition of the solubilized drugs, with focus on CrEL and paclitaxel, and of concomitantly administered drugs. The ability of the surfactants to form micelles in aqueous solution as well as biological fluids (e.g. plasma) appears to be of great importance with respect to the pharmacokinetic behavior of the formulated drugs. Due to drug entrapment in the micelles, plasma concentrations and clearance of free drug change significant leading to alteration in pharmacodynamic characteristics. We conclude with some perspectives related to further investigation and development of alternative methods of administration. PMID- 11392449 TI - Oral delivery of taxanes. AB - Oral treatment with cytotoxic agents is to be preferred as this administration route is convenient to patients, reduces administration costs and facilitates the use of more chronic treatment regimens. For the taxanes paclitaxel and docetaxel, however, low oral bioavailability has limited development of treatment by the oral route. Preclinical studies with mdr1a P-glycoprotein knock-out mice, which lack functional P-glycoprotein activity in the gut, have shown significant bioavailability of orally administered paclitaxel. Additional studies in wild type mice revealed good bioavailability after oral administration when paclitaxel was combined with P-glycoprotein blockers such as cyclosporin A or the structurally related compound SDZ PSC 833. Based on the extensive preclinical research, the feasibility of oral administration of paclitaxel and docetaxel in cancer patients was recently demonstrated in our Institute. Co-administration of cyclosporin A strongly enhanced the oral bioavailability of both paclitaxel and docetaxel. For docetaxel in combination with cyclosporin A an oral bioavailability of 90% was achieved with an interpatient variability similar to that after intravenous drug administration; for paclitaxel the oral bioavailability is estimated at approximately 50%. The safety of the oral route for both taxanes is good. A phase II study of weekly oral docetaxel in combination with cyclosporin A is currently ongoing. PMID- 11392450 TI - Population pharmacokinetics and pharmacokinetic-pharmacodynamic relationships for docetaxel. AB - The population approach has been implemented prospectively in the clinical development of docetaxel (Taxotere). Overall 640 patients were evaluable for the population PK/PD analysis. The PK analysis evidenced significant covariates explaining the inter-patient variability of docetaxel clearance and the PK/PD analysis demonstrated that the variability in clearance was a significant predictor of several safety endpoints. In patients with clinical chemistry suggestive of mild to moderate liver function impairment (SGOT and/or SGPT > 1.5 x ULN concomitant with alkaline phosphatase >2.5 x ULN), total body clearance was lowered by an average of 27%. Specific safety analyses demonstrated that these patients are at a significantly higher risk than others for the development of severe docetaxel-induced side effects. Population PK/PD data were fully integrated into the regulatory dossier and in the labeling of docetaxel worldwide. Population PK/PD models are being used to elaborate a simulation model to predict the survival of patients with non-small cell lung cancer treated with docetaxel. PMID- 11392453 TI - Cellular correlates of olfactory learning in the rat piriform cortex. AB - This review describes research that combines cellular physiology with behavioral neuroscience, to study the cellular mechanisms underlying learning and memory in the mammalian brain. Rats were trained with an olfactory conditioning paradigm, in which they had to memorize odors in order to be rewarded with drinking water. Such training results in rule learning, which enables enhanced acquisition of odor memory. Training results in the following learning-related physiological modifications in intrinsic and synaptic properties in olfactory (piriform) cortex pyramidal neurons: 1. increased neuronal excitability, indicated by reduced afterhyperpolarization, and 2. increased synaptic transmission, indicated by reduced paired-pulse facilitation. These modifications are correlated to enhanced learning capability rather than to storage of memory for specific odors. In addition, using a different paradigm of odor-training, it is shown that NMDA and betra-adrenergic receptors are involved at different stages of long-term memory consolidation. PMID- 11392451 TI - Body surface area as a determinant of pharmacokinetics and drug dosing. AB - Body surface area (BSA) was introduced into medical oncology in order to derive a safe starting dose for phase I studies of anticancer drugs from preclinical animal toxicology data. It is not clear however, as to why dosing by BSA was extended to the routine dosing of antineoplastic agents. Several formulas exist to estimate BSA, but the formula derived by DuBois and DuBois is the one used in adult medical oncology. This formula was derived based on data from only nine patients; subsequent attempts to validate the formula have found the DuBois formula to either over or underestimate the actual determined BSA. While cardiac output does correlate with BSA, the relationship between BSA and other physiologic measures relevant for drug metabolism and disposition, such as, renal and hepatic function, is weak or nonexistent. Further only epirubicin, etoposide, and carboplatin have been studied to determine if dosing by BSA would reduce interpatient variability, and none of these drugs were found to have significant relationships between their pharmacokinetics and BSA. Future clinical trials of new agents should not presume that dosing based on BSA reduces interpatient variability. Studies should examine the role, if any, BSA has in dosing new chemotherapeutic agents in initial phase I studies. PMID- 11392454 TI - Long-term depression: a cellular basis for learning? AB - Long-term depression (LTD) comprises a persistent activity-dependent reduction in synaptic efficacy which typically occurs following repeated low frequency afferent stimulation. Hippocampal LTD has been a subject of particular interest due to the established role of the hippocampus in certain forms of information storage and retrieval. Recently, it was reported that LTD in the CA1 region may be associated with novelty acquisition in rats. CA1 LTD expression may also be increased in stressful conditions. This suggests a more complex role for this form of plasticity than the oft-cited hypothesis that it simply serves to prevent synapse saturation, by means, for example, of enabling reversal of long-term potentiation (LTP). One possibility is that LTD may be directly involved in the creation of a memory trace. Alternatively, LTD may prime a synapse in readiness for the expression of LTP, thereby contributing indirectly to information storage. There is increasing evidence that LTD is not mechanistically the reverse of LTP. Although some common processes exist, molecular, biochemical, electrophysiological and pharmacological studies all point to several quite distinct induction and maintenance mechanisms for this form of synaptic plasticity. Taken together these findings suggest that hippocampal LTD must be considered in a new light. This review focuses on the interpretation of novel and established information with regard to LTD in the hippocampal CA1 region in terms of its possible role as a cellular basis for learning and memory. PMID- 11392452 TI - Drug interactions of paclitaxel and docetaxel and their relevance for the design of combination therapy. AB - The taxanes' interaction with other anticancer drugs have been extensively investigated in in vitro and in animal models as well as in humans due to the outstanding antitumor activity in a broad range of malignancies. Paclitaxel and docetaxel are endowed of a rich and complex pharmacology whereby different pharmacodynamic effects are observed depending on the sequence of their administration in respect with the companion drug, and the type of drug that is combined. Pharmacokinetic interference is often but not always a basis of the pharmacodynamic effect. In addition, the vehicle of clinical formulation, especially Cremophor EL for paclitaxel, influence the pharmacological effect. Finally, new interaction based on as yet unknown mechanisms drive the two taxanes to multiple additive/synergistic relationships with new signal transduction drugs, such as modulators of the epidermal-growth-factor family of receptors and farnesyl-transferase inhibitors. The ongoing effort to better understanding such a rich pharmacology is worth continuing in view of designing new and better combinations of the taxanes. PMID- 11392455 TI - Drug dependence as a disorder of neural plasticity: focus on dopamine and glutamate. AB - Drug addiction, as a disease, has grown to reach the level of a social illness. Psychostimulants, opiates, alcohol, nicotine and cannabis abuse affects millions worldwide and virtually all classes of modern society. In spite of the enormous proportions of its spread, intimate neurobiological mechanisms leading to distintictive features of this pathological status, such as craving for the abused substance and loss of control over intake, remain largely obscure and pharmacotherapies sadly unsatisfactory. In the last decade, preclinical and clinical research in this field has made great progress to improve our understanding of the brain mechanisms which form the basis of this illness. The review of recent literature, which represents the focus of the present paper, leads to the emerging consensus that an alteration of physiological mechanisms of neural plasticity within the brain dopamine and glutamate systems may underlie some of the behavioral abnormalities occurring during the dependence cycle. In particular, a reduction of dopamine neuronal activity and glutamate neurotransmission at the level of the ventrotegmental area, after withdrawal from chronic administration of drugs of abuse, may work in concert with alterations in other forebrain areas, such as the nucleus accumbens and the amygdaloid complex. In addition, following prolonged periods of abstinence, even after somatic withdrawal signs have vanished, responsiveness of these systems to drugs of abuse remains abnormal. This suggests that these two neurotransmitters may play a substantial role in the long-lasting, enduring changes typical of the addictive process and may represent ideal targets for pharmacological intervention aimed at normalizing forms of neural plasticity impaired after chronic drug intake. PMID- 11392456 TI - Thalamocortical synaptic connections: efficacy, modulation, inhibition and plasticity. AB - The thalamic input to the neocortex is communicated by glutamatergic synapses. The properties and organization of these synapses determine the primary level of cortical processing. Similar to intracortical synapses, both AMPA and NMDA receptors in young and mature animals mediate thalamocortical transmission. Kainate receptors participate in thalamocortical transmission during early development. The shape of thalamocortical synaptic potentials is similar to the shape of intracortical potentials. On the other hand, thalamocortical synapses have on average a higher release probability than intracortical synapses, and a much higher number of release sites per axon. As a result, the transmission of each thalamocortical axon is significantly more reliable and efficient than most intracortical axons. Thalamic axons specifically innervate a subset of inhibitory cells, to create a strong and secure feed-forward inhibitory pathway. Thalamocortical connections display many forms of synaptic plasticity in the first postnatal week, but not afterwards. The implications of the functional organization of thalamocortical synapses for neocortical processing are discussed. PMID- 11392457 TI - Is reaching eye-centered, body-centered, hand-centered, or a combination? AB - There are currently three main views on the neural basis of visually guided reaching: 1) neurons in the superior parietal lobe guide arm movements in a spatial framework that is centered on the body; 2) neurons in the intraparietal sulcus guide arm movements in a spatial framework that is centered on the eye; 3) neurons in the caudal part of premotor cortex guide arm movements in a spatial framework that is centered on the arm and hand. The three viewpoints are mutually compatible and may fit into a larger pattern. Eye-centered representations of target position, and body-centered representations of arm and hand position, may be integrated to form a hand-centered representation close to the output stage in caudal premotor and primary motor cortex. PMID- 11392458 TI - High resolution SPECT in small animal research. AB - Single Photon Emission Computed Tomography (SPECT) is a technique used to assess physiological and biochemical processes under in vivo conditions. SPECT generates tomographic images from blood flow, glucose metabolism and receptor characteristics using radioactively labelled substances. This paper reviews the state of the art of in vivo imaging of laboratory animals in modified human and dedicated animal SPECT scanners. SPECT cameras with special collimators currently reach spatial resolutions up to 1 mm and sensitivities of about 1000 cps/MBq, allowing observation of receptor activity concentration changes in the pico-mole range. The time resolution of such cameras strongly depends on the pharmacological behaviour of the tracer and can range from several minutes to hours. Within these limits the functional characterization of many processes is possible. SPECT also offers the possibility to set up dynamic study protocols and repeated measurements of the same animal. This technique reduces the need for sacrificing animals, as was commonly practiced before the development of animal cameras. Animal SPECT gives the opportunity to monitor physiological and biochemical processes in animals in vivo, without interfering with the system under observation, and may become a valuable adjunct to the instrumentation (autoradiography, in vitro methods) of animal research. PMID- 11392459 TI - The role of neurotrophic factors in psychostimulant-induced behavioral and neuronal plasticity. AB - Several neurotrophic factors influence the development, maintenance and survival of dopaminergic neurons in the mammalian central nervous system (CNS), including neurotrophin-3 (NT-3), brain derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF), basic fibroblast growth factor (bFGF) and glial derived neurotrophic factor (GDNF). This review focuses on the role of these neurotrophic factors in psychostimulant-induced behavioral sensitization, a form of dopamine-mediated neuronal plasticity that models aspects of paranoid schizophrenia as well as drug craving among psychostimulant addicts. Whereas NT 3, CNTF and bFGF appear to play a positive role in psychostimulant-induced behavioral sensitization, GDNF inhibits this form of behavioral plasticity. The role of BDNF in behavioral sensitization, however, remains elusive. While it has been shown that neurotrophic factors can influence the behavioral, structural and biochemical phenomena related to psychostimulant-induced neuronal plasticity, it is unclear which neurotrophic factors are important physiologically and which have purely pharmacological effects. In either case, examining the role of neurotrophic factors in behavioral sensitization may enhance our understanding of the mechanisms underlying the development of paranoid psychosis and drug craving and lead to the development of novel pharmacological treatments for these disorders. PMID- 11392460 TI - Maternal cocaine administration during pregnancy induces apoptosis in fetal rat heart. AB - We have previously demonstrated that cocaine induces apoptosis in primary cultures of fetal rat cardiomyocytes. The current study was designed to determine whether cocaine administered to the mother during pregnancy induced apoptosis in fetal rat heart. Pregnant rats were treated with cocaine subcutaneously (30 and 60 mg/kg per day) starting at day 15 of gestation and were terminated at day 21. Cocaine produced a dose-dependent increase in apoptotic cell death in the fetal heart by 1.3-fold (30 mg/kg per day) and 2.4-fold (60 mg/kg per day) of the control level (1.99+/-0.15%). Cocaine-induced DNA fragmentation in the fetal heart showed characteristic apoptotic ladder. In accordance, cocaine dose dependently increased activities of caspase-3, caspase-8, and caspase-9 in the fetal heart by 0.5-, 0.6-, and 0.6-fold, respectively, at 30 mg/kg per day, and by 3.3-, 2.9-, and 2.3-fold, respectively, at 60 mg/kg per day. In contrast, cocaine showed no effect on caspase activities in the maternal heart. Bcl-2 and Bax proteins were detected in fetal rat heart, with 2.2-fold higher expression of Bcl-2 than Bax. Cocaine significantly increased Bax protein levels and decreased Bcl-2 protein levels, leading to a 7.5-fold increase in the Bax-to-Bcl-2 ratio in fetal rat heart. We conclude that cocaine causes apoptosis in fetal rat heart in vivo by upregulating the Bax-to-Bcl-2 ratio and increasing caspase activities, which is likely to play an important role in the adverse effects of cocaine on heart development. PMID- 11392461 TI - Inhibition of T-type and L-type calcium channels by mibefradil: physiologic and pharmacologic bases of cardiovascular effects. AB - Ca2+ channel antagonists of the dihydropyridine, benzothiazepine, and phenylalkylamine classes have selective effects on L-type versus T-type Ca2+ channels. In contrast, mibefradil was reported to be more selective for T-type channels. We used the whole-cell patch-clamp technique to investigate the effects of mibefradil on T-type and L-type Ca2+ currents (I(CaT) and I(CaL)) recorded at physiologic extracellular Ca2+ in different cardiac cell types. At a stimulation rate of 0.1 Hz, mibefradil blocked I(CaT) evoked from negative holding potentials (HPs) (-100 mV to -80 mV) with an IC50 of 0.1 microM in rat atrial cells. This concentration had no effect on I(CaL) in rat ventricular cells (IC50: approximately3 microM). However, block of I(CaL) was enhanced when the HP was depolarized to -50 mV (IC50: approximately 0.1 microM). Besides a resting block, mibefradil displayed voltage- and use-dependent effects on both I(CaT) and I(CaL). In addition, inhibition was enhanced by increasing the duration of the step-depolarizations. Similar effects were observed in human atrial and rabbit sinoatrial cells. In conclusion, mibefradil combines the voltage- and use dependent effects of dihydropyridines and benzothiazepines on I(CaL). Inhibition of I(CaL), which has probably been underestimated before, may contribute to most of the cardiovascular effects of mibefradil. PMID- 11392462 TI - Can moderate alcohol intake limit the size of myocardial infarction? AB - Several observational and epidemiologic analyses suggest that moderate drinking of alcohol (usually two drinks per day) may have beneficial effects on the heart. Specifically it may reduce the incidence of ischemic heart disease and improve the outcome after a myocardial infarction. The purpose of this study was to test the hypothesis that long-term, moderate consumption of alcohol has a direct cardioprotective effect, reducing myocardial infarct size in rats subjected to ischemia/reperfusion. Two groups of rats were given alcohol in their drinking water at concentrations of 15% or 36%; control rats received water (n = 10 for all groups). After 16 weeks of treatment, the rats were anesthetized and subjected to 60 min of coronary artery occlusion and reperfusion. The ischemic risk region (% LV) was not significantly different among groups. Infarct size (% risk zone) was 49+/-7%, 44+/-4%, and 50+/-8%, respectively (p = 0.79). Thus no significant differences in infarct size between control and alcohol-fed rats were observed. There was no correlation between alcohol level and infarct size in alcohol-treated rats (r = 0.20, p = 0.45). Arterial pressures were lower in the 36% group at baseline and throughout the study compared with the control and 15% groups, but heart rates were similar in all groups. Moderate consumption of alcohol failed to alter infarct size compared with control and thus it is unlikely that alcohol drinking has a direct myocardial-protecting effect on the outcome of ischemia and reperfusion. PMID- 11392463 TI - Na+/H+ exchange inhibitor SM-20220 attenuates leukocyte adhesion induced by ischemia-reperfusion. AB - Leukocytes play a key role in ischemia-reperfusion-induced tissue injuries. It has been suggested that blocking the Na+/H+ exchanger improves ischemic injuries such as stroke. In this study, we investigated the effect of the Na+/H+ exchanger inhibitor SM-20220 (N-[aminoiminomethyl]- 1-methyl-1H-indole-2-carboxamide methanesulfonate) on leukocyte-endothelial cell interactions during ischemia reperfusion. SM-20220 (0.3-1.0 mg/kg i.v.) given after ischemia significantly attenuated the leukocyte adhesion in the mesenteric postcapillary venules that was induced by transient superior mesenteric artery occlusion. At 60 min after reperfusion, the numbers of adherent leukocytes in groups treated with vehicle or SM-20220 (0.3 mg/kg) were 15.1+/-2.9 cells/100 microm/3 min and 3.0+/-0.7 cells/100 microm/3 min (p < 0.01), respectively. In a transient middle cerebral artery occlusion model, i.v. infusion of SM-20220 (0.4 mg/kg per hour) for 1 h, beginning 1 h after the start of occlusion, significantly reduced both the infarct size and the increase in brain myeloperoxidase activity, compared with the vehicle group (p < 0.01 and p < 0.05, respectively). In summary, this is the first evidence that the leukocyte adhesion to the endothelium that is induced by ischemia-reperfusion is attenuated by the inhibition of Na+/H+ exchanger activity in vivo. Our results suggest that Na+/H+ exchanger inhibitors may prevent ischemia-reperfusion injuries such as stroke partly through the attenuation of leukocyte-endothelial cell interactions. PMID- 11392464 TI - Bucindolol, a nonselective beta 1- and beta 2-adrenergic receptor antagonist, decreases beta-adrenergic receptor density in cultured embryonic chick cardiac myocyte membranes. AB - Bucindolol and carvedilol, nonselective beta1- and beta2-adrenergic receptor antagonists, have been widely used in clinical therapeutic trials of congestive heart failure. The aim of the current study was to investigate long-term effects of bucindolol or carvedilol on beta-adrenergic receptor protein and gene expression in cardiac myocytes. Embryonic chick cardiac myocytes were cultured and incubated with bucindolol (1 microM), carvedilol (1 microM), or norepinephrine (1 microM) for 24 h. 125I-iodocyanopindolol binding assays demonstrated that incubation with norepinephrine or bucindolol, but not carvedilol, significantly decreased beta-adrenergic receptor density in crude membranes prepared from the myocytes. Neither bucindolol nor carvedilol significantly stimulated adenylyl cyclase activity in membranes from drug untreated cells. Unlike by norepinephrine, the receptor density reduction by bucindolol incubation was not accompanied by a change in beta1-adrenergic receptor messenger RNA abundance. A decrease in membrane beta-adrenergic receptor density without a change in cognate messenger RNA abundance was also observed in hamster DDT1 MF2 cell line incubated with bucindolol (1 microM, 24 h). We conclude that incubation with bucindolol, but not carvedilol, results in true reduction of beta-adrenergic receptor density in chick cardiac myocyte membranes by mechanisms that are distinct from those responsible for receptor density reduction by the agonist norepinephrine. PMID- 11392465 TI - Dose-dependent blockade of the angiotensin II type 1 receptor with losartan in normal volunteers. AB - Losartan, an angiotensin II type 1 receptor (AT1) antagonist, was developed as a more specific alternative to angiotensin-converting enzyme (ACE) inhibitors. At a daily dose of 50 mg, losartan is currently evaluated in large outcome trials involving patients with hypertension and postmyocardial infarction. The current study evaluated the level and duration of blockade of a pressor response to angiotensin II by 50 and 150 mg of losartan, compared with 32 mg of candesartan. Eight normotensive volunteers were randomly assigned to a single dose of losartan 50 or 150 mg, candesartan 32 mg, or placebo. Subjects were re-randomized after a 2-week washout period to complete all four study arms. Radial artery systolic pressure response to exogenous angiotensin II was measured at 2, 6, 12, and 24 h after administration of drug. Losartan 50 mg reduced the pressure response to exogenous angiotensin II significantly only at 6 h. In contrast, candesartan and losartan 150 mg produced a greater reduction in the pressure response to angiotensin II throughout the 24-h period. This suppression was not paralleled by a reduction in resting systemic arterial pressure. Higher doses than 50 mg of losartan might be evaluated to elicit optimal clinical effects. PMID- 11392467 TI - Homocysteine reduces smooth muscle [Ca2+]i and constrictor responses of isolated arterioles. AB - Chronic elevation of plasma homocysteine concentration has been shown to be associated with impaired vascular function. The acute direct effect of homocysteine on the tone and vasoactive responses of arterioles and the possible underlying mechanisms, however, have not yet been elucidated. Thus arterioles were isolated from gracilis muscle of rats (d: approximately 130 microm) and their diameter was measured by videomicroscopy. Homocysteine (10(-6)-10(-4) M) elicited dose-dependent dilation of arterioles (maximum: 44+/-6% at 10(-4) M). The dilation was not affected by the presence of the nitric oxide synthase inhibitor Nomega-nitro-L-arginine methyl ester or by removal of the endothelium, or the free radical scavenger catalase and superoxide dismutase, or the K+ channel inhibitors glibenclamide, 4-aminopyridine, or tetraethyl ammonium. Incubation of vessels with homocysteine (10(-4) M, 20 min) did not affect dilations to acetylcholine or sodium nitroprusside, whereas it significantly decreased constrictions to norepinephrine (at 10(-6) M; control: 57+/-7%, homocysteine: 21+/-5%) and to the thromboxane A2 analogue U46619 (at 10(-8) M: control: 44+/-3%, homocysteine: 20+/-4%). Homocysteine (10(-4) M), similar to the voltage-operated Ca2+ channel inhibitor nitrendipine (10(-8) M), significantly decreased the arteriolar smooth muscle [Ca2+]i as assessed by changes in the fura 2 ratiometric signal (R(Ca), -6+/-1% and -24+/-3%, respectively). These data suggest that in isolated arterioles homocysteine decreases pressure-induced tone and responses to vasoconstrictor agents, likely by altering Ca2+ signaling of arteriolar smooth muscle. PMID- 11392466 TI - Effect of gestational age, corticosteroids, and birth on expression of prostanoid EP receptor genes in lamb and baboon ductus arteriosus. AB - The aim of this study was to determine the effect of corticosteroids, gestational age, and birth on the expression of genes encoding prostanoid receptors in the lamb and baboon ductus arteriosus. The ductus arteriosus was obtained from 34 lambs and eight baboons, including chronically instrumented fetuses of both species exposed to either corticosteroid or vehicle. Expression of prostanoid receptor genes was quantified using Northern blot analysis relative to each of two housekeeping genes. Expression of both the EP3 and EP4 receptor genes was detected in lamb ductus and the level of expression of both genes was unaffected by corticosteroids. Expression of the EP4 receptor gene was lower in the ductus obtained from term lambs compared with preterm lambs and was lower still in neonatal animals, whereas no variation was observed in EP3 receptor gene expression. Expression of the EP4 receptor gene was also confirmed in fetal baboon ductus arteriosus, and maternal administration of corticosteroid did not reduce EP4 receptor gene expression in the baboon. We conclude that advancing gestational age and birth may inhibit prostaglandin E2-mediated relaxation of the ductus through a corticosteroid-independent reduction in EP4 receptor gene expression. PMID- 11392469 TI - Effects of ACE inhibition on left ventricular failure and oxidative stress in Dahl salt-sensitive rats. AB - Dahl salt-sensitive (DS) rats fed high-salt diet exert compensated left ventricular (LV) hypertrophy and eventually develop heart failure. Oxidative stress has been shown to be involved in myocardial remodeling and failure and thus might play an important role in this transition from hypertrophy to failure. We measured the amount of reactive oxygen species (ROS) in the myocardium from DS rats by using electron spin resonance spectroscopy with 4-hydroxy-2,2,6,6 tetramethyl-piperidine-N-oxyl (hydroxy-TEMPO) and also examined the effects of chronic angiotensin-converting enzyme (ACE) inhibition on the transition. We divided DS rats (5 weeks old, 150-200 g) into three groups: low-salt (0.3% NaCl) diet for 10 weeks (LS group), high-salt (8% NaCl) diet for 10 weeks (HS-10+V group), and high-salt diet and cilazapril (10 mg/kg body weight per day) started after 5 weeks of high-salt diet and maintained for 5 weeks (HS-10+Cil group). Systolic blood pressure (mm Hg) was significantly elevated in the HS-10+V (229+/ 5) and HS-10+Cil (209+/-5) groups compared with the LS group (141+/-2). The amount of myocardial ROS was not changed after 5 weeks of high-salt diet, but significantly increased in HS-10+V rats compared with LS rats, and was abolished in the HS-10+Cil group. HS-10+V rats exerted the clinical signs of heart failure, including increased lung weight and pleural effusion, associated with LV hypertrophy and LV cavity dilatation. In the HS-10+Cil group, signs of heart failure were significantly attenuated despite only a modest reduction in systolic blood pressure (-20 mm Hg). The progression of LV failure after hypertrophy in high-salt-loaded DS hypertensive rats was associated with increased myocardial ROS, and ACE inhibitor could prevent this transition from compensated hypertrophy to failure. PMID- 11392468 TI - Cicletanine stimulates nitric oxide release and scavenges superoxide in endothelial cells. AB - Cicletanine ((+/-)3-(4-chlorophenyl)-1,3-dihydro-7-hydroxy-6-methylfuro-[3,4-c] pyridine) 3-(4-chlorophenyl)-1,3-dihydro-7-hydroxy-6-methylfuro-[3,4-c] pyridine) is a novel antihypertensive vasodilator with an incompletely understood mechanism of action. In the studies described here, the release of nitric oxide and superoxide (O2-) stimulated by cicletanine was measured simultaneously in the endothelium of isolated rat aortic rings. Highly sensitive electrochemical nitric oxide and O2- microsensors were placed near the surface of endothelial cells and the kinetics of nitric oxide and O2- release were monitored in situ. The response times for nitric oxide and O2- microsensors were 100 micros and 50 micros, respectively, and detection limit was 10(-9) M. Cicletanine stimulated nitric oxide release in aorta endothelium at (micromolar) therapeutic concentrations that were consistent with the concentrations of the compound to induce endothelium-dependent vasorelaxation in isolated rat aorta. The peak concentration of nitric oxide was 160+/-8 nM. This concentration was about 70% and was 60% lower as compared with the nitric oxide peak concentration observed after stimulation with receptor-independent agonist (calcium ionophore A23187) and receptor-dependent agonist (acetylcholine), respectively. However, after administration of cicletanine, only a small concentration of O2- was recorded (peak 3.1+/-0.2 nM) contrary to a large concentration (27+/-1.35 nM) observed after stimulation with A23187). Cicletanine not only stimulated nitric oxide release but also was a potent scavenger of O2- at nanomolar level. Both of these effects may contribute to potent vasorelaxation properties of cicletanine and its long-term therapeutic actions, resulting in cardiovascular tissue protection. PMID- 11392470 TI - CGP 41251, a new potential anticancer drug, improves contractility of rat isolated cardiac muscle subjected to hypoxia. AB - The aim of the present work was to examine the effects of 4'-N-benzoyl staurosporine (CGP 41251), a protein kinase C inhibitor with broad antiproliferative activity in many cell lines, on the rat isolated heart contractility under normoxic and hypoxic conditions. Additionally, we examined the effects of CGP 41251, WB-4101 (alpha1a -adrenoceptor antagonist), chloroethylclonidine (CEC) (alpha1b-adrenoceptor antagonist) and selective damage of endocardial endothelium by Triton X-100 on the protection against hypoxia induced by preconditioning of rat heart tissue. Experiments were performed on rat isolated left ventricular papillary muscle. The following parameters were measured: force of contraction (Fc), velocity of contraction (+dF/dt) and velocity of relaxation (-dF/dt). The temperature of the bath solution was 37 degrees C +/- 0.5 degrees C, and rate of electrical stimulation was 0.5 Hz. At concentrations less than 1 microM CGP 41251 did not cause any changes in contractility of rat heart. At 1 and 3 microM, significant positive inotropic action was observed. Treatment of rat papillary muscle by CGP 41251 at 3 microM reduced decreasing of contractility by simulated hypoxia and reperfusion. Moreover, protective effects of preconditioning was not affected by addition of CGP 41251 neither at 1 nor at 3 microM. Pretreatment with CEC at 3 microM, and selective damage of endocardial endothelium induced by fast (1-s) immersion of papillary muscle in 0.5% Triton X-100, but not pretreatment with WB-4101, abolished the protective effects of preconditioning. The results imply that CGP 41251 improves contractility of heart muscle under normoxic and hypoxic conditions, and does not alter hypoxic preconditioning in rat isolated cardiac tissue. Moreover, it was shown that alpha1b-adrenoceptors and endocardial endothelium are involved in triggering of preconditioning in rat isolated heart muscle. PMID- 11392471 TI - Taurine modulates arginine vasopressin-mediated regulation of renal function. AB - Taurine has been implicated in the regulation of arginine vasopressin (AVP) secretion, and we have previously shown altered renal excretory function in the taurine-depleted rat. To further elucidate the role of taurine in AVP-mediated renal responses, the effects of an antagonist for renal AVP receptors were examined in four groups of conscious rats: control, taurine-supplemented, taurine depleted, and taurine-repleted. Control and taurine-supplemented rats displayed similar and significant AVP receptor antagonist-induced elevations in fluid excretion, sodium excretion, and free water clearance but a marked reduction in urine osmolality. These effects are consistent with inhibition of endogenous AVP activity. By contrast, in the taurine-depleted rats, the magnitude and the time course of drug-induced renal excretory responses lagged behind those of the control and taurine-supplemented groups. Further, baseline urine osmolality was significantly higher in the taurine-depleted compared with the control or taurine supplemented groups. However, after administration of the antagonist, taurine depleted rats manifested a delayed but more marked reduction in urine osmolality, thereby eliminating the baseline differential that existed between the taurine depleted rats and control or taurine-supplemented groups. Consistent with these observations, plasma AVP was significantly increased in the taurine-depleted compared with the control rats. Interestingly, taurine repletion shifted all responses closer to the control group. Analysis of the data suggests that the effect of the antagonist on renal excretory function is related primarily to altered tubular reabsorption activity. These observations suggest that taurine modulates renal function, and, thereby, body fluid homeostasis, through an AVP dependent mechanism. PMID- 11392472 TI - Evidence for the involvement of phosphatidylinositol 3-kinase in fMLP-stimulated neutrophil adhesion to ICAM-1-transfected cells. AB - Phosphatidylinositol 3-kinase (PI-3K) controls important intracellular steps involved in inflammation, immunity, and cell growth. PI-3K also modulates leukocyte integrin adhesiveness. In this study we evaluated the role of PI-3K on neutrophil adhesion to intercellular adhesion molecule-1 (ICAM-1)-transfected cells. N-formyl-methionyl-leucyl-phenylalanine (fMLP)-stimulated neutrophil adhesion was inhibited by wortmannin and LY294002, two unrelated PI-3K inhibitors, whereas phorbol myristate acetate (PMA)-induced neutrophil adhesion was not inhibited by them. After fMLP stimulation, a rapid activation of AKT and ERK was observed. However, only activation of AKT was reversed by the PI-3K inhibitors. Neutrophil expression of the beta2-integrins Mac-1, lymphocyte function-associated antigen-1(LFA-1), and gp150.95 was not affected by wortmannin, nor was expression of the activation epitope recognized by MAB24. We conclude that (a) PI-3K is involved in fMLP-activated neutrophil adhesion to ICAM 1-transfected cells, (b) the mechanism involved is not mediated by the modulation of beta2-integrin expression or activation, and (c) another mechanism seems to involve the adhesion to ICAM-1 when a cellular system of adhesion is used. PMID- 11392473 TI - Effect of atorvastatin on plasminogen activator inhibitor type-1 synthesis in human monocytes/macrophages. AB - The fibrinolytic inhibitor plasminogen activator inhibitor type 1 (PAI-1) plays a role in the development of atherothrombosis and is produced by macrophages that infiltrate the atherosclerotic vessel wall. Because statins are effective in reducing atherosclerosis, we investigated if they modulate the synthesis of PAI-1 in human monocytes/macrophages. To this end, we studied the effect of atorvastatin in different models of monocyte/macrophage differentiation, such as differentiated human promyelocytic cell line HL-60 and human peripheral blood monocyte-derived macrophages. HL-60 cells were differentiated along monocyte lineage by phorbol myristate acetate (PMA) or a mixture of transforming growth factor-beta type 1 (TGF-beta1)/1alpha,25-dihydroxyvitamin D3 (D3). In these conditions, PAI-1 synthesis was strongly induced and atorvastatin upregulated this synthesis, especially during TGF-beta1/D3-induced differentiation. Recombinant human tumor necrosis factor-alpha (TNF-alpha) strongly upregulated PAI-1 synthesis in PMA- or TGF-beta1/D3-differentiated cells, and the potentiating effect of atorvastatin was of the same order as in the absence of TNF-alpha. Mevalonate reversed the enhancing effect of atorvastatin. In mature human monocyte-derived macrophages, atorvastatin, alone or in combination with TNF-alpha, TGF-beta1, or PMA, did not exert any significant effect on PAI-1 synthesis. Basal production of urokinase (uPA), which was below detection limits in HL-60 cells and very low in human monocyte-derived macrophages, was not altered by atorvastatin. These results show that atorvastatin upregulates PAI-1 synthesis during the early stages of monocyte/macrophage differentiation, but has no effect on PAI-1 and uPA synthesis in mature human monocyte-derived macrophages. Atorvastatin did not significantly interact with the upregulating action of TNF-alpha on PAI-1 synthesis during differentiation. PMID- 11392474 TI - Anti-ischemic potential of drugs related to the renin-angiotensin system. AB - Actions mediated by the renin-angiotensin system may be inhibited at various levels: renin itself may be inhibited, angiotensin-I (A-1) conversion to angiotensin-II (A-II), or binding of A-II at the A-II type 1 (A-II1) receptor. The angiotensin-converting enzyme (ACE) inhibitors and the A-II1 receptor antagonists are now clinically established. Because ACE is a relatively unspecific peptidase which catalyses the breakdown of A-I, bradykinin and neuropeptides like substance P and neurotensin, the effects of ACE inhibitors go far beyond the prevention of A-II production. On the other hand, in certain tissues like vascular and cardiac tissue, A-II is produced by other enzymes, for instance chymase, and ACE inhibitors do not consistently prevent A-II production. The action of A-II1 receptor antagonists may also not be confined to prevention of binding of A-II at the A-II1 receptor, as by rebound more A-II may bind at the A-II type 2 (A-II2) receptor and thus mediate until now not well defined effects. Thus, anti-ischemic actions of these drugs may be related to multiple mechanisms. Inhibition of A-II effects at the A-II1 receptor may prevent systemic and coronary vasoconstriction and growth effects of A-II on various cell types. In addition, A-II may potentiate, by pre- and postsynaptic mechanisms, activation of the sympathetic nervous system. Prevention of breakdown of bradykinin, substance P and neurotensin may result in direct vasodilation or release of nitrous oxide from the endothelium. Thus, growth-inhibiting effects may also be mediated. All these mechanisms seem to direct to a reduction of cardiac load by vasodilation and to a limitation of cardiovascular cell growth. While the systemic circulating renin-angiotensin system is probably responsible for control of cardiac load, local systems seem to control cell growth. Systemic effects seem to depend on activation of the renin-angiotensin system which has been shown in various ischemic syndromes. Activation of various components of the renin-angiotensin system has been demonstrated in myocardial ischemia, acute myocardial infarction and coronary occlusion and reperfusion models as well as in chronic left ventricular dysfunction post-myocardial infarction. While animal models of stress induced myocardial ischemia have revealed predominantly positive results, clinical studies, which mostly were small and not well controlled, were equivocal. Large clinical trials with ACE inhibitors in acute myocardial infarction showed small benefits over placebo. Hypotension seems to be a critical side-effect in this situation. Experimental models show protective effects of both ACE inhibitors and A-II1 receptor antagonists in the situation of ischemia and reperfusion. New data on large clinical trials in patients at risk of cardiovascular events but normal left ventricular function demonstrate clear benefits of an ACE inhibitor. Large clinical trials in patients with chronic left ventricular dysfunction post-myocardial infarction show reduction of ischemic events. PMID- 11392475 TI - Vascular protective effects of angiotensin converting enzyme inhibitors and their relation to clinical events. AB - Endothelial cells are a rich source of a variety of vasoactive substances, which either cause vasodilation or vasoconstriction. Important endothelium-derived vasodilators are prostacyclin, bradykinin, nitric oxide and endothelium-derived hyperpolarizing factor. In particular, nitric oxide inhibits cellular growth and migration. In concert with prostacyclin. nitric oxide exerts potent anti atherogenic and thromboresistant properties by preventing platelet aggregation and cell adhesion. Endothelium-derived contracting factors include the 21 amino acid peptide endothelin (ET). vasoconstrictor prostanoids such as thromboxane A2 and prostaglandin H2, as well as free radicals and components of the renin angiotensin system. In hypertension, elevated blood pressure transmits into cardiovascular disease by causing endothelial dysfunction. Hence, modem therapeutic strategies in human hypertension focus on preserving or restoring endothelial integrity. Angiotensin converting enzyme (ACE) inhibitors are a primary candidate for that concept as they inhibit the circulating and local renin angiotensin system. Angiotensin converting enzyme is an endothelial enzyme which converts angiotensin-I (A-I) into angiotensin-II (A-II). This effect of the ACE inhibitor prevents direct effects of angiotensin-II such as vasoconstriction and proliferation in the vessel wall but also prevents activation of the ET system and of plasminogen activator inhibitor. Furthermore, inhibition of ACE prolongs the half-life of bradykinin and stabilizes bradykinin receptors linked to the formation of nitric oxide and prostacyclin. In isolated arteries ACE inhibitors prevent the contractions induced by angiotensin II and enhance relaxation induced by bradykinin. Chronic treatment of experimental hypertension with ACE inhibitors normalizes endothelium-dependent relaxation to acetylcholine and other agonists. In addition, the dilator effects of exogenous nitric oxide donors are enhanced, at least in certain models of hypertension. In humans with essential hypertension ACE inhibitors augment endothelium-dependent relaxation to bradykinin, while those to acetylcholine remain unaffected, at least in the time frame of the published studies, i.e. 3-6 months. In patients with coronary artery disease, however, paradoxical vasoconstriction to acetylcholine is markedly reduced after 6 months of ACE inhibition. After myocardial infarction ACE inhibitors reduce the development of overt heart failure, the occurrence of reinfarction and cardiovascular death in hypertensive patients. These effects have also been demonstrated in a subgroup analysis of the SOLVD (Studies of Left Ventricular Dysfunction) trial. Thus, in summary, ACE inhibitors are an important class of drugs providing cardiovascular protection in patients with increased cardiovascular risk. PMID- 11392476 TI - Anti-ischemic effects of angiotensin-converting enzyme inhibitors: a future therapeutic perspective. AB - The renin-angiotensin system and angiotensin-converting enzyme (ACE) are increasingly being implicated in the pathogenesis of coronary artery disease and its sequelae and the potential of ACE inhibitors to protect the heart is a topic that has emerged recently as a matter for scientific discussion. Experimental and clinical studies have shown the beneficial effects of ACE inhibitors on the metabolism, function and structure of healthy and damaged hearts and these data support the concept of both primary and secondary cardioprotection with this class of drugs. Animal studies have demonstrated the potential beneficial effects of ACE inhibition at a variety of sites, including improvement of endothelial function, inhibition of platelet aggregation, prevention of atherosclerotic lesions and inhibition of myointimal proliferation, extending the concept to a more general definition of cardiovascular protection with ACE inhibitors involving both the heart and the vessels. ACE inhibitors prevent stimulation of smooth muscle cell angiotensin-II (A-II) receptors, thereby blocking both contractile and proliferative actions of A-II. In addition, ACE inhibition of kininase inhibits the breakdown of bradykinin, a direct stimulant of nitric oxide release from the intact endothelial cell. Thus, at the cellular level ACE inhibition shifts the balance of ongoing mechanisms in favour of those promoting vasodilatory, anti-aggregatory, antithrombotic and antiproliferative effects. Although these data have not all been validated in human studies, the reduction of ischemic events in studies of ACE inhibition in left ventricular dysfunction (LVD) and, more recently, also in patients without LVD, cannot be explained solely by improved hemodynamics, and it is possible that actions on the endothelium, the atherosclerotic process and platelets are at least in part responsible. So, the available data underlie the potential benefits of ACE inhibition in the field of ischemic heart disease and atherosclerosis; the results of ongoing studies in humans looking more directly at the influence of ACE inhibitors in this setting are awaited with interest. PMID- 11392477 TI - Beyond remodeling: a new paradigm for angiotensin-converting enzyme inhibitors following myocardial infarction. AB - Angiotensin converting enzyme (ACE) inhibitors, originally designed to treat hypertension, were quickly demonstrated to confer hemodynamic and survival benefit to patients with congestive heart failure. Extending this paradigm to patients with left ventricular dysfunction (LVD) post-myocardial infarction (MI), ACE inhibitors were shown to attenuate ventricular remodeling and reduce mortality. An unexpected finding that ACE inhibitors could reduce the incidence of myocardial infarction, prompted enormous interest in their anti-ischemic potential. Indeed, a significant body of experimental literature supports the concept that ACE inhibitors have direct anti-atherosclerotic and anti-ischemic effect. Recent clinical trials have shown that ACE inhibitors confer significant protection from ischemic events. PMID- 11392478 TI - Renin-angiotensin system inhibition improves coronary flow reserve in hypertension. AB - Angiotensin II not only elevates arterial pressure, it adversely alters hemodynamics and cardiovascular structure and exacerbates the course of hypertensive disease. Alterations in coronary hemodynamics, including reductions in coronary blood flow and flow reserve, reflect the pathophysiology of arteriolar disease and associated endothelial dysfunction thereby promoting coronary insufficiency and increasing overall cardiovascular risk. In spontaneously hypertensive rats, coronary flow reserve, the difference between basal coronary blood flow and the flow achieved during maximal coronary vasodilation achieved by physiological or pharmacological interventions, is drastically impaired at rest; however, it can be improved significantly by pharmacological agents that inhibit the renin-angiotensin system, alone or in combination. The combination of an angiotensin-converting enzyme (ACE) inhibitor and an angiotensin-II (type 1) receptor blocker, in equidepressor doses, markedly improved coronary flow reserve to levels seen in normotensive Wistar Kyoto rats after 12 weeks of treatment, while diminishing cardiovascular mass and improving systemic hemodynamics and ventricular metabolic demands. These findings suggest the potential merits for clinical studies employing the combination of ACE inhibitor and angiotensin receptor blocker therapy in patients with hypertension and hypertensive heart disease. PMID- 11392479 TI - Randomly amplified polymorphic DNA (RAPD) profiles for the distinction of Lactobacillus species. AB - Forty-one type and reference strains of Lactobacillus were evaluated using their randomly amplified polymorphic DNA band profiles. Developed bands for each strain were distinct and enabled discrimination. The best correlations were obtained applying the Pearson product moment correlation coefficient (r) together with the unweighted pair group method using arithmetic averages algorithm. All of the strains were clearly differentiated at and below the 72% similarity value. Species discrimination might be possible making use of the distinctly polymorphic bands amplified specific to a strain. PMID- 11392480 TI - Signal recognition particle mediated protein targeting in Escherichia coli. AB - The signal recognition particle (SRP) is a conserved ribonucleoprotein complex that binds to targeting sequences in nascent secretory and membrane proteins. The SRP guides these proteins to the cytoplasmic membrane in prokaryotes and the endoplasmic reticulum membrane in eukaryotes via an interaction with its cognate receptor. The E. coli SRP is relatively small and is currently used as a model for fundamental and applied studies on translation-linked protein targeting. In this review recent advances in our understanding of the structure and function of the E. coli SRP and its receptor are discussed. In particular, the interplay between the SRP pathway and other targeting routes, the role of guanine nucleotides in cycling of the SRP and the substrate specificity of the SRP are highlighted. PMID- 11392481 TI - Colonization patterns of wood-inhabiting fungi on baits in Hong Kong rivers, with reference to the effects of organic pollution. AB - The diversity of wood-inhabiting fungi was investigated by submerging woody baits at upstream and downstream sites of the Lam Tsuen and Tai Po Rivers in Hong Kong. The diversity of fungi in the Lam Tsuen River was also compared with that on natural woody substrates found in a previous study. There were differences in the species composition between the upstream and downstream sites, possibly reflecting natural variations along the river. The Tai Po River downstream was organically polluted, which appeared to have little effect on species diversity since more species were recorded. Organic pollution may, however, cause a shift in species composition. The fungal communities on baits and natural substrates in the Lam Tsuen River were similar, although a lower diversity was observed on baits. This may be related to the period of submergence and the fact that a single wood type was used. Cercophora spp. occurred frequently downstream in the Tai Po River, while the common species in the Lam Tsuen River were Aquaticola rhomboidea and Pseudoproboscispora aquatica. Further interpretation on the effects of organic pollution was limited because of single collection data but appropriate experimental designs--putting baits in unimpacted sites for assessing human impacts in streams--are suggested. PMID- 11392482 TI - Effects of gaseous NO2 on cells of Nitrosomonas eutropha previously incapable of using ammonia as an energy source. AB - Cells of Nitrosomonas eutropha grown under anoxic conditions with hydrogen as electron donor and nitrite as electron acceptor were initially unable to oxidize ammonia (ammonium) and hydroxylamine when transferred to oxic conditions. Recovery of ammonia and hydroxylamine oxidation activity was dependent on the presence of NO2. Under oxic conditions, without addition of NO2, ammonia consumption started after 8 - 9 days, and small amounts of NO and NO2 were detectable in the gas atmosphere. Removing these nitrogen oxides by intensive aeration, ammonia oxidation activity decreased and broke off after 15 days. Addition of gaseous NO2 (25 ppm1) led to a fast recovery of ammonia oxidation (3 days). Simultaneously, the arrangement of intracytoplasmic membranes (ICM) changed from circular to flattened vesicles, the protein pattern revealed an increase in the concentration of a 27 and a 30 kDa polypeptide, and the cytochrome c content increased significantly. PMID- 11392483 TI - The terminal inverted repeats of the linear plasmid SCP1 of Streptomyces coelicolor A3(2) possess a truncated copy of the transposon Tn4811 of Streptomyces lividans 66. AB - Streptomyces coelicolor A3(2), the best genetically studied streptomycete and Streptomyces lividans 66 are very closely related strains. This is further emphasized by our finding that a truncated copy of Tn4811 of S. lividans is present in the terminal inverted repeats of the S. coelicolor giant linear plasmid SCP1. The copy of Tn4811 in SCP1 lacks the first 1276 bp and shows only minor changes in the nucleotide sequence of the remaining 4.12 kb. Tn4811 exists in both ends of SCP1. PMID- 11392484 TI - Assessment of the genetic diversity of Xylella fastidiosa isolated from citrus in Brazil by PCR-RFLP of the 16S rDNA and 16S-23S intergenic spacer and rep-PCR fingerprinting. AB - The genetic diversity among twenty three strains of Xylella fastidiosa, isolated from sweet orange citrus, was assessed by RFLP analysis of the 16S rDNA and 16S 23S intergenic spacer and by rep-PCR fingerprinting together with strains isolated from coffee, grapevine, plum and pear. The PCR products obtained by amplification of the 16S rDNA and 16S-23S spacer region were digested with restriction enzymes and a low level of polymorphism was detected. In rep-PCR fingerprinting, a relationship between the strains and their hosts was observed by using the BOX, ERIC and REP primers. Two major groups were obtained within the citrus cluster and relationships to the geographic origin of the strains revealed. Citrus strains isolated from the States of Sao Paulo and Sergipe formed one group and strains from the Southern States formed another group. Distinct origins of X. fastidiosa in the Southern and Southeastern States is postulated. The pear isolate was distantly related to all of the other X. fastidiosa strains. PMID- 11392485 TI - Identification and expression of the Pseudomonas syringae pv. aptata hrpZ(Psa) gene which encodes an harpin elicitor. AB - A sequence homologous to an internal fragment 0.75 kb BstXI of the Pseudomonas syringae pv. syringae hrpZ gene was identified in Pseudomonas syringae pv. aptata NCPPB 2664, the causal agent of bacterial blight in sugar beet, lettuce and other plants. and in E. coli DH10B (pCCP1069) containing the P. syringae pv. aptata hrp gene cluster. PCR with oligonucleotides, based on the hrpZ(Pss) gene and used as primers with the total genomic DNA of P. syringae pv. aptata, amplified a 1 kb fragment that hybridized with the probe in highly stringent conditions. The amplicon was cloned into the pGEM-T plasmid vector, amplified in E. coli DH5alpha and sequenced. The sequence showed 95%, 83% and 61% identity with those of hrpZ(Pss), hrpZ(Psg and hrpZ(Pst) genes encoding the harpins of the P. syringae pv. syringae, glycinea and tomato, respectively. The amplicon was cloned into the pMAL expression system. The expressed protein, fused with maltose-binding protein, was cleaved with a specific protease factor Xa, and purified using affinity chromatography. On the basis of the amino acid sequence and its ability to induce HR in tobacco leaves, it was identified as a P. syringae pv. aptata harpin. PMID- 11392486 TI - Ascobotryozyma americana gen. nov. et sp. nov. and its anamorph Botryozyma americana, an unusual yeast from the surface of nematodes. AB - A new teleomorphic genus Ascohotryozyma, with a single species, A. americana, is proposed. Its anamorph is a Botryozyma that differs from the type species, B. nematodophila, on distributional, physiological, and molecular criteria; it is described as Botryozyma americana, anam. sp. nov. Ascobotryozyma is characterized by globose asci bearing four lunate ascospores. Fusion of thallus cells precedes ascus formation. Ascobotryozyma americana was isolated from the surface of nematodes (Panagrellus dubius) associated with galleries of the poplar borer (Saperda calcarata) in trembling aspen (Populus tremuloides) in eastern Washington, USA. The teleomorph has not been produced in pure culture. PMID- 11392487 TI - Changes in fatty acid composition and degree of unsaturation of (brady)rhizobia as a response to phases of growth, reduced water activities and mild desiccation. AB - The effects of growth phase, reductions in the water activity (a(w)) of the growth medium and mild desiccation on the composition and the degree of unsaturation of cellular fatty acids (CFA) of Sinorhizobium meliloti, Bradyrhizobium elkanii and Bradyrhizobium japonicum were studied. During the course of growth, an interchange of cis-vaccenic with lactobacillic acid and a slight increase in palmitic acid were observed while other fatty acids remained constant. The degree of unsaturation was significantly higher in the exponential phase of growth. Reductions in the a(w) of the medium led to an increase in lag phase, a reduction in growth rate and maximal optical densities (OD) in stationary phase cells. A decrease in the degree of unsaturation of CFA was also observed as the a(w) was reduced from 0.999 to 0.969 and after desiccation to 83.5% relative humidity (R.H.). The changes in the degree of unsaturation of CFA observed after growth at reduced a(w) may be one of the pre-adaptation steps to endure more severe desiccation. PMID- 11392488 TI - Citrate efflux in glucose-limited and glucose-sufficient chemostat culture of Penicillium simplicissium. AB - Citrate excretion by Penicillium simplicissimum was investigated in a chemostat. Carbon-limited grown P. simplicissimum did not excrete no citrate. Citrate was excreted, however, when growth was nitrogen-limited. Further effects of nitrogen limitation were a slightly increased rate of glucose and oxygen consumption. This behaviour is typical for a so-called 'overflow metabolism', i.e. the uncoupling of anabolism from catabolism under conditions of carbon excess. Still more citrate was excreted by nitrogen-limited P. simplicissimum when (i) the extracellular osmolarity was increased from 0.2 to 1.5 osm kg(-1) or (ii) when the pH was increased from 4 to 7; or (iii) when the extracellular potassium concentration was lowered from 6 to 0.5 mM. These results were interpreted in terms of a higher energy-consumption under these conditions. PMID- 11392489 TI - Microbiology and physiology of Cachaca (Aguardente) fermentations. AB - Cachaca (aguardente) is a rum-style spirit made from sugar cane juice by artisanal methods in Brazil. A study was made of the production, biochemistry and microbiology of the process in fifteen distilleries in Sul de Minas. Identification of 443 yeasts showed Saccharomyces cerevisiae to be the predominant yeast but Rhodotorula glutinis and Candida maltosa were predominant in three cases. Bacterial infection is a potential problem, particularly in older wooden vats, when the ratio of yeasts:bacteria can be 10:1 or less. A study of daily batch fermentations in one distillery over one season in which 739 yeasts were identified revealed that S. cerevisiae was the predominant yeast. Six other yeast species showed a daily succession: Kluyveromyces marxianus, Pichia heimii and Hanseniaspora uvarum were present only at the beginning, Pichia subpelliculosa and Debaryomyces hansenii were detected from mid to the end of fermentation, and Pichia methanolica appeared briefly after the cessation of fermentation. Despite a steady influx of yeasts from nature, the species population in the fermenter was stable for at least four months suggesting strong physiological and ecological pressure for its maintenance. Cell densities during the fermentation were: yeasts - 4 x 108/ml; lactic acid bacteria -4 x 10(5)/ml; and bacilli - 5 x 10(4)/ml. Some acetic acid bacteria and enterobacteriaceae appeared at the end. Sucrose was immediately hydrolysed to fructose and glucose. The main fermentation was complete after 12 hours but not all fructose was utilised when harvesting after 24 hours. PMID- 11392490 TI - Analysis of yeasts derived from natural fermentation in a Tokaj winery. AB - The diversity of yeast flora was investigated in a spontaneously fermenting sweet white wine in a Tokaj winery. The non-Saccharomyces yeasts dominating the first phase of fermentation were soon replaced by a heterogeneous Saccharomyces population, which then became dominated by Saccharomyces bayanus. Three Saccharomyces sensu stricto strains isolated from various phases of fermentation were tested for genetic stability, optimum growth temperature, tolerance to sulphur dioxide, copper and ethanol as well as for the ability to produce hydrogen sulphide and various secondary metabolites known to affect the organoleptic properties of wines. The analysis of the single-spore cultures derived from spores of dissected asci revealed high stability of electrophoretic karyotypes and various degrees of heterozygosity for mating-types, the fermentation of galactose and the production of metabolic by-products. The production levels of the by-products did not segregate in a 2:2 fashion, suggesting that the synthesis of these compounds is under polygenic control. PMID- 11392491 TI - The National Service Framework on coronary heart disease: is it sufficiently evidence-based? PMID- 11392492 TI - Faecal fat: time to give it up. PMID- 11392493 TI - Medullary thyroid carcinoma: role of genetic testing and calcitonin measurement. AB - All patients with a thyroid nodule should have their plasma CT measured. Stimulated CT is generally better than basal, but in the lower ranges false negatives and false positives still occur. In families with hereditary MTC, RET gene mutation analysis has superseded measurement of plasma CT in the detection of asymptomatic disease gene carriers. All individuals with apparently sporadic MTC, but in whom there is some suspicion of familial disease, should also have RET genetic analysis. A negative DNA result practically excludes the possibility of hereditary MTC in families where an index case has been investigated and obviates the need for further biochemical evaluation. Disease gene carriers may be divided into three distinct risk groups depending on the specific RET gene mutation in the family. The age at which presymptomatic surgery has to be performed depends on the risk group to which the patient belongs. Compared with the results of DNA analysis, the results of CT stimulation tests have become less important in the assessment of timing of surgery. During follow-up of patients who underwent surgery, measurement of plasma basal CT is still useful. The high sensitivity of measuring stimulated CT levels does not outweigh the burden of life-long periodic stimulation tests and the limited clinical consequences of slightly elevated levels. Stimulation tests are inevitable for persons at risk who prefer not to have genetic testing. PMID- 11392494 TI - Clinical investigation and management of patients with renal stones. PMID- 11392495 TI - Pre- and post-analytical factors that may influence use of serum prostate specific antigen and its isoforms in a screening programme for prostate cancer. PMID- 11392496 TI - Automated immunoassay analysers. PMID- 11392497 TI - Effect of incorrect gestational dating on Down's syndrome and neural tube risk assessment. AB - Down's syndrome risks are estimated between 15 and 20 completed weeks' gestation (cGW) using an algorithm involving maternal age and serum alpha-fetoprotein (AFP), chorionic gonadotrophin and unconjugated oestriol levels, each expressed as a multiple of the median level (MoM) at the cGW. The AFP MoM itself is the basis for screening for open neural tube defects (oNTD). Because medians change during this period, gestational dating must be accurate so that appropriate medians are used. A calculated Down's syndrome risk > 1:380 at term is generally considered to indicate a 'high-risk' pregnancy. This study focused on 378 patients with reported risk < or = 1:500 based on physician-supplied cGW (and hence considered at 'low risk' for Down's syndrome) to determine the effect of common 1-2-week dating errors on risk estimates. Using the original analytical data, each patient's risk was recalculated for each week over the 15-20 weeks, and classified into three categories: < 1:380 'low'; 1:380-1:100 'moderate'; and > 1:100 'high'. Advancing originally 'low-risk' patients by one week increased the risk by 1.09-14.1 times (median 3.18, mean 3.60); 46 (12.2%) became 'moderate' and 2 (0.5%) became 'high' risk. Advancing by two weeks increased risks 1.58-60.5 times (median 10.03, mean 12.04); 131 (36.5%) became 'moderate' and 39 (10.9%) became 'high' risk. Predictably, oNTD screening results also were affected. Although 1-2 week differences in AFP medians had little effect on most patients in this study sample, some who originally were oNTD negative became oNTD positive, whereas others who had been oNTD positive became screen negative. Thus, in many cases, a 1-2 week dating error may have only minimal effect on the estimated risks for chromosome or neural tube defects, but in other cases the effect of such an error would be significant. PMID- 11392498 TI - Multiple sclerosis: use of light-chain typing to assist diagnosis. AB - Although the presence of oligoclonal IgG with abnormal kappa/lambda light-chain ratio in multiple sclerosis (MS) has been known for many years, this finding has not been put to diagnostic use in most routine clinical laboratories. In a retrospective study we report differences in the oligoclonal banding patterns between multiple sclerosis and non-MS patients. We had sufficient cerebrospinal fluid (CSF) on 36 from 71 patients with oligoclonal bands for immunofixation for kappa and lambda light chains, and for free kappa and free lambda. Thirteen out of 14 patients with clinically confirmed MS had predominantly IgG (kappa) banding. In contrast, in seven out of eight patients with diagnoses other than MS the IgG was linked to both kappa and lambda light chains in approximately equal proportions. Nine out of 14 patients with probable/possible/suspected MS showed predominantly IgG (kappa) banding; five others in this group had both IgG (kappa) and IgG (lambda) and free lambda light chains. The finding of IgG (kappa) bands in CSF samples with oligoclonal bands supports a diagnosis of MS. PMID- 11392499 TI - 3-Hydroxyanthranilic acid, an L-tryptophan metabolite, induces apoptosis in monocyte-derived cells stimulated by interferon-gamma. AB - 3-Hydroxyanthranilic acid (3-HAA), a metabolite of L-tryptophan, accumulates in monocyte-derived cells (THP-1), but not in other cell lines tested (MRC-9, H4, U373MG, Wil-NS), following immune stimulation that induces indoleamine-2,3 dioxygenase (IDO), a rate-limiting enzyme in the L-tryptophan kynurenine pathway. We examined whether metabolites of the L-tryptophan-kynurenine pathway act to induce apoptosis in monocytes/macrophages. Of the L-tryptophan metabolites tested, only 3-HAA at a concentration of 200 micromol/L was found to induce apoptosis in THP-1 and U937 cells. The addition of ferrous or manganese ions further enhanced apoptosis and free radical formation by 3-HAA in these two types of cells. The apoptotic response induced by 3-HAA was significantly attenuated by the addition of antioxidant, alpha-tocopherol or Trolox (a water-soluble analogue of vitamin E), and the xanthine oxidase inhibitor, allopurinol. In addition, the 3-HAA-induced apoptotic response was slightly attenuated by catalase, but not by superoxide dismutase (SOD), indicating that generation of hydrogen peroxide is involved in this response. Interferon-gamma (IFN-gamma), an inducer of IDO, potently induced apoptosis in THP-1 cells, but not in U937 cells, in the presence of ferrous or manganese ions. This different susceptibility to apoptosis inducer between THP-1 and U937 cells may depend on the capacity of the cells for 3-HAA synthesis following IDO induction by IFN-gamma. Furthermore, apoptosis was suppressed by cycloheximide in THP-1 cells, suggesting that newly synthesized proteins may be essential for apoptotic events. These results suggest that 3-HAA induces apoptosis in monocytes/macrophages under inflammatory or other pathophysiological conditions. PMID- 11392500 TI - Effect of storage of plasma and serum on enzymatic determination of non esterified fatty acids. AB - Many contradictory results have been published on the stability of total non esterified fatty acids in blood, plasma and serum under different storage conditions. The present study was undertaken to investigate the stability of non esterified fatty acids, measured with an enzymatic method, in samples of EDTA treated plasma and serum under different temperature conditions. We conclude that EDTA-treated plasma and serum can both be used for analysis. Specific reference values should be established depending on the type of sample chosen. Samples that cannot be analysed immediately can be stored at -20 degrees C for at least 14 days without significant changes in the concentration of total non-esterified fatty acids. None of the other storage conditions and periods studied are suitable for the measurement of non-esterified fatty acid concentration. PMID- 11392501 TI - Investigation of applicability of a mid-infrared spectroscopic method using an attenuated total reflection accessory and a new near-infrared transmission method for determination of faecal fat. AB - In many laboratories, the titrimetric method of Van de Kamer is used for the analysis of faecal fat content of patients suspected of steatorrhoea. We investigated the applicability of a mid-infrared (MIR) spectroscopic method, using an attenuated total reflection (ATR) accessory, and a new near-infrared (NIR) spectroscopic method. For the NIR method, sealed plastic bags containing the stool samples were used as transmission cells. Standardization was obtained using a previously described MIR method, with a NaCl flow-cell, as reference method. Partial least-squares regression was used for the calibration of each method. Full cross-validation of the calibration set was used for the internal validation of each method. Fifteen per cent of the stool samples could not be estimated with the ATR method within reasonable accuracy limits compared with the reference. The standard error of prediction of the NIR method was 1.1 g/dL. We conclude that the new NIR method is a promising technique for routine use. However, further experiments need to be done with triplicate measurements of each sample and the use of an external validation set. PMID- 11392502 TI - Fragile X syndrome in Calcutta, India. AB - Fragile-X-linked mental retardation usually results from amplification of the CGG repeat in the 5' untranslated region of the FMR1 gene. To assess the extent of variation of the CGG repeat in the population from the eastern region of India we studied 98 mentally retarded individuals living in and around Calcutta and identified 21 distinct alleles ranging in size from 8 to 44 CGG repeats. A repeat size of 28 was the most frequent; this value is different from the most frequent repeat size found in other studies, indicating a racial or ethnic variation. Patients with the clinical features of the syndrome have been found to carry expanded CGG repeats. Thus, it can be inferred that the expansion of CGG repeats may be a frequent cause of the syndrome in our population. PMID- 11392503 TI - External glucose meter quality assurance in the UK: are we doing enough? PMID- 11392504 TI - Haem arginate interferes with estimation of carcinoembryonic antigen. PMID- 11392505 TI - Interference by danazol with the Porter-Silber method for determination of urinary 17-hydroxycorticosteroids. PMID- 11392506 TI - Troponin T as a first-line test: the Royal Liverpool and Broadgreen University Hospital experience. AB - Many hospital laboratories are unable to offer a cardiac troponin service because of the cost of providing this assay in addition to existing cardiac enzyme profiles: we circumvented this problem by withdrawing the conventional cardiac enzyme service and substituting cardiac troponin T. By ensuring that only one specimen for cTnT is analysed per episode of chest pain. substantial financial savings have been achieved. PMID- 11392507 TI - Evaluation of glucose meters in neonatal unit. PMID- 11392508 TI - Provision of interpretive comments on biochemical report forms. PMID- 11392509 TI - Cardiac marker testing following surgery. PMID- 11392510 TI - Stability of intact parathyroid hormone in blood samples. PMID- 11392511 TI - Effect of inflammatory response on trace element and vitamin status. PMID- 11392512 TI - Another hazard in plasma aluminium assay by graphite furnace atomic absorption. PMID- 11392513 TI - Neurologic aspects of adenylosuccinate lyase deficiency. AB - Adenylosuccinate lyase deficiency is an autosomal-recessive disorder of the purine de novo synthesis pathway, diagnosed up to now in approximately 40 patients. The clinical presentation is characterized by severe neurologic involvement including seizures, developmental delay, hypotonia, and autistic features. Neonatal seizures and a severe infantile epileptic encephalopathy are often the first manifestations of this disorder. The existence of genetic heterogeneity for the adenylosuccinate lyase defect could account for variability of the clinical presentation. Deficiency of purine nucleotides, impairment of energy metabolism, and toxic effects are potential mechanisms of cerebral damage. Laboratory investigations show the presence in urine and cerebrospinal fluid of succinylpurines, which are normally undetectable. Currently, no effective treatment is available for adenylosuccinate lyase deficiency. A search for this disorder should be included in the screening program of children with unexplained neonatal seizures or severe infantile epileptic encephalopathy. PMID- 11392514 TI - Early cognitive and communication development in children with focal brain lesions. AB - Early cognitive and language development of children with congenital focal brain lesions, documented by magnetic resonance imaging, was studied in 18 cases, 9 with left-hemisphere damage and 9 with right-hemisphere damage, at about 2 (Time 1) and 4 years of age (Time 2). All of the children showed normal cognitive development, but their global Griffiths Developmental Scales scores were lower at Time 2, and developmental profiles across individual subscales revealed side specific effects, resembling the adult left/right cerebral hemisphere lesion model. Expressive lexicon and grammar were delayed, more often in left-hemisphere damaged than in right-hemisphere-damaged children, at Time 1 and Time 2. Functional findings were not related to the size and location of the brain lesion, whereas the presence of epilepsy was a highly significant predictor of cognitive and language outcome, irrespective of the side of the lesion. The stable disadvantage in the verbal domain shown by left-hemisphere-damaged children within the age range of this study might suggest that the left hemisphere has some initial bias for language learning. The effects of right hemisphere damage were more variable and emerged at a later stage of language development. PMID- 11392515 TI - Definite and suspected multiple sclerosis in children: long-term follow-up and magnetic resonance imaging findings. AB - Twenty-five children at the ages of 3 to 18 years with an initial diagnosis of acute disseminated encephalomyelitis were followed in the Clinic of Child Neurology for a period of 2 to 8 years. In 10 children, there were data for clinically definite or laboratory-supported definite multiple sclerosis. The other 15 children in our study were considered as having suspected multiple sclerosis. Brain magnetic resonance imaging (MRI) performed in 15 children disclosed multiple hyperintense lesions on T2-weighted imaging in 13 children: 10 with definite multiple sclerosis and 3 with suspected multiple sclerosis. The clinical manifestations did not always correspond to the size and location of the MRI lesions of demyelination. Follow-up revealed normalization of the neurologic examination in 18 patients (72%) and abnormal neurologic findings in 7 patients (28%) (6 children with definite multiple sclerosis and 1 with suspected multiple sclerosis). Magnetic resonance imaging follow-up in children with definite multiple sclerosis disclosed a reduction in the size of the lesions in 3; enlargement or new lesions were established in the other 7 cases, and 2 cases were without clinical signs of new attacks. Correlation was done concerning the findings of the cerebrospinal fluid examination, transcranial magnetic stimulation, evoked potentials, computed tomography, and MRI. The role of MRI for an early diagnosis of multiple sclerosis in children is discussed. The dynamic follow-up of the pathologic changes is of prognostic significance for the course of the disease that could be a definite cessation of the process in acute disseminated encephalomyelitis cases or transition to multiple sclerosis. PMID- 11392516 TI - Brain activity and cognitive status in pediatric patients: development of a clinical assessment protocol. AB - The purpose of this study was to test the validity of a new computerized task to assess children's cognitive problem-solving skills using the brain event-related potentials. This event-related potential-computerized cognitive problem-solving task does not require a child to give a verbal or motor (ie, pointing) response. The event-related potential waveforms were recorded from 20 typically developing children. Two nonverbal, problem-solving tasks (tasks 1 and 2) were developed for each of two age groups (5 and 6 years). For each task, single pictures, taken from an existing standardized test of nonverbal problem solving, were individually and sequentially presented on a computer screen. One of the seven pictures was classified as incongruent or outside category; it did not belong with the other pictures. As predicted, the event-related potential amplitudes were significantly larger to the outside- versus within-category pictures. This effect was found for tasks 1 and 2 for the 5- and 6-year-old children. Children as young as 5 years of age reliably exhibit brain activity, which can be used to infer cognitive problem-solving skill. This assessment paradigm may eventually serve as a clinically useful adjunct to a thorough neurologic and neurodevelopmental assessment of selected pediatric populations, such as those presenting with moderate-severe cerebral palsy whose expressive language and motor skills are notably impaired. PMID- 11392517 TI - Occurrence of Rett syndrome in boys. AB - The neurologic disorder Rett syndrome was originally described exclusively in girls. We present two boys with clinical features of Rett syndrome. Other than head circumference deceleration, no longer considered mandatory, patient 1 meets all of the criteria. Using fluorescent in situ hybridization analysis, 97.6% of cells were found to be karyotypically normal (46,XY). No mutation was detected on screening of the coding region of the MECP2 gene. The second patient also has classic features of Rett syndrome. However, cytogenetic analysis of peripheral blood revealed a karyotype 47,XXY[23]/46,XY[7] confirming mosaicism for Klinefelter's syndrome. A T158M missense mutation in the methylcytosine-binding domain of the MECP2 gene was identified. A diagnostic bias against the clinical identification of Rett syndrome in boys may exist. This presentation of the male phenotype could be more common than it would appear, although boys with MECP2 mutations might also manifest in other ways. Rett syndrome remains a clinical diagnosis that should not be dismissed in boys, and thorough evaluation including karyotype and mutation testing is warranted. PMID- 11392518 TI - Ictal single photon emission computed tomography in absence seizures: apparent implication of different neuronal mechanisms. AB - Absence seizures represent a complex group of epilepsy, characterized by lapse of consciousness with staring. Bilateral, synchronous, and symmetric bursts of 3-Hz spike-and-wave discharges are observed on the electroencephalogram, whereas interictal background activity is normal. This kind of epilepsy has to be differentiated from other generalized epilepsies such as juvenile absence epilepsy and juvenile myoclonic epilepsy. Moreover, absence seizures, together with generalized spike-and-wave discharges, may coexist with other types of epilepsy such as frontal lobe epilepsy, temporal lobe epilepsy, benign epilepsy with centrotemporal spikes, and childhood epilepsy with occipital paroxysms. We have carried out ictal single photon emission computed tomography (SPECT) in 10 patients with clinical evidence of absence seizures with the aim to better understand and to distinguish this kind of seizure as primarily or secondarily generalized to a specific area and to obtain more information on the neuronal mechanisms involved in the different types of seizures, usually not identifiable at the first appearance. During the long follow-up period (9 months to 14 years), 7 of the 10 examined patients underwent interictal SPECT when they became seizure free. Our data permitted, in two patients, the diagnosis of childhood absence seizures; in three patients, they suggested the possibility of later appearance of other seizure types, on the basis of focal hyperperfusion indicating a possible focal firing. In three of the examined patients, the diagnosis of idiopathic localization-related epilepsies mimicking childhood absence seizures could be performed. In the last two patients, the hypothesis of a coexistence of absences with partial and generalized seizures was considered. From our results, it can be presumed that ictal SPECT findings may contribute to the physiopathologic classification of the different types of epilepsies. Moreover, anticonvulsant treatment more appropriate to the different forms of seizures can be used. PMID- 11392519 TI - Clinical evaluation of conversational speech fluency in the acute phase of acquired childhood aphasia: does a fluency/nonfluency dichotomy exist? AB - Traditional neurologic tenets claim that the clinical picture of acquired childhood aphasia is nonfluent irrespective of lesion location. In the past 20 years, however, several case studies have shown that fluent aphasic patterns can be observed in children with acquired childhood aphasia. But the question remains open as to whether the pattern of their speech characteristics is similar to the one described in adult aphasics as studies addressing spontaneous speech fluency characteristics in larger series of children with acquired childhood aphasia are scarce. The objective of this study was to investigate whether an analysis of spontaneous speech fluency as has previously been performed in adult aphasics by other investigators would also yield two distinct groups of aphasic children and, if so, whether the distribution of the different speech characteristics in both groups would reflect the rank order found in adults, that is, whether nonfluent verbal output characteristics would predominate in one group and fluent features in the other. Audiotaped and videotaped recordings of 24 cooperative children with acute acquired childhood aphasia unselected for age, gender, etiology, and aphasia severity ratings were analyzed according to 10 different speech characteristics. A cluster analysis (two-means clustering) was performed to seek the existence of two distinct groups of aphasic children. Results were confirmed, and exact P values were computed with Mann-Whitney U-tests. A two-means clustering created two distinct classes. Mann-Whitney U-tests ranked the speech characteristics according to their discriminating power between clusters. Comparing this rank order with the one previously found in adults revealed a high correlation (Spearman's rank correlation: r = .915, P << .005), thus indicating that the clusters we found were highly similar to the adult clusters. Thus, the use of the speech variables proposed to evaluate adult aphasic spontaneous speech enabled us to demonstrate a fluent/nonfluent dichotomy in a childhood aphasic population as well. This study shows that the traditional views on the uniformity of the clinical picture of acquired childhood aphasia are obsolete. Our findings corroborate data issued from several case reports of fluent acquired childhood aphasia and from the few studies focusing on speech fluency in acquired childhood aphasia, which all point to the existence of an adultlike heterogeneity of childhood aphasic syndromes. Current clinical evidence no longer supports the hypotheses of equipotentiality and progressive lateralization but favors the notion that the anatomic substrate for language representation in the child is similar to that in adults, even in young subjects. PMID- 11392520 TI - Isabelle Rapin: an autobiography. PMID- 11392522 TI - Recurrence in pituitary adenomas in childhood and adolescence. AB - Pediatric pituitary adenomas are thought to behave more aggressively than their adult counterparts, and the ability to predict the degree of such behavior remains elusive. Proliferation marker Ki-67 and tumor suppressor gene p53 mutations have been used in adults to assist in the evaluation of invasiveness and recurrence; however, their use in childhood and adolescence remains anecdotal. Our study evaluates the proliferative potential in pituitary adenomas of five patients and its relationship with recurrence or persistence of endocrinologic or clinical abnormalities. For such assessment, tissues were stained with monoclonal antibodies BP53-12 forp53 tumor suppressor gene mutation and MIB-1, which binds to cell cycle-specific nuclear antigen Ki-67. In our series, one patient with recurrent adenoma demonstrated the highest (50%) p53 immunoreactivity. Ki-67-stained nuclei ranged from 0 to 2%, failing to identify the recurrent tumor. Therefore, p53 immunoreactivity, rather than Ki-67 nuclear stain, may be useful for identification of recurrent pituitary adenomas in childhood and adolescence. PMID- 11392521 TI - A study of novel polymorphisms in the upstream region of vasoactive intestinal peptide receptor type 2 gene in autism. AB - We investigated the vasoactive intestinal peptide receptor type 2 (VIPR2) gene as a candidate gene for autism. We searched for mutations in the VIPR2 gene in autistic individuals, and 10 novel polymorphisms were identified. Three polymorphisms in the upstream region were studied in detail, and there was no significant difference in the frequencies between the autistic group (n = 14) and unrelated controls (n = 52). The distribution of the genotypes in two of the three polymorphisms differed somewhat between autistic subjects with gastrointestinal problems and those without. Moreover, there was a trend showing a correlation between the genotypes for the third polymorphism and the severity of stereotypical behavior as ranked by the Gilliam Autism Rating Scale. These preliminary results suggest that VIPR2 may have a role in gastrointestinal symptoms and stereotypical behaviors in autism, although a larger collection of samples suitable for transmission disequilibrium tests is necessary to validate the results. PMID- 11392523 TI - Effect of Antiepileptic drugs on plasma lipoprotein (a) and other lipid levels in childhood. AB - Antiepileptic drugs may alter plasma lipid status in epileptic patients. We conducted a study to assess the effect of phenobarbital, carbamazepine, and valproate on plasma levels of lipoprotein (a), total cholesterol, triglycerides, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, apolipoprotein A, and apolipoprotein B in 22 epileptic children. The children were separated as group 1, seven children, mean age 1.6+/-0.2 years, treated with phenobarbital, 5 mg/kg/day, twice daily; group 2, seven children, mean age 9.8+/ 1.2 years, treated with carbamazepine, 20 mg/kg/day, twice daily; and group 3, eight children, mean age 6.8+/-0.6 years, treated with valproate, 20 mg/kg/day, twice daily. Plasma lipoprotein (a) and other lipid levels were studied before (pretreatment) and at 3 and 6 months of treatment. Friedman two-way analysis of variance and Wilcoxon's signed-rank test were used for statistical analysis, and the results were expressed as the mean and standard error of the mean. The mean age of children in group 1 was significantly low, compared with groups 2 and, 3 (P < .001). The mean pretreatment lipid levels between the groups were not significant. The increase in lipoprotein (a) at 3 and 6 months and high-density lipoprotein cholesterol at 6 months was statistically significant in group 1 (P < .025). We suggest a careful monitoring of plasma levels of lipoprotein (a) and other lipids in epileptic children treated with antiepileptic drugs. PMID- 11392524 TI - Progressive cardiac dysautonomia observed in patients affected by classic Rett syndrome and not in the preserved speech variant. AB - Incidence of sudden death in Rett syndrome is greater than that of the general population, and cardiac electrical instability is a prime suspect cause. The objective of the present study was the evaluation of heart rate variability, a marker of autonomic activity, in females affected by classic Rett syndrome and atypical variants for a possible explanation of the higher risk for sudden death observed in these subjects. Our study showed that girls with classic Rett syndrome had significantly lower heart rate variability and longer corrected QT intervals than in atypical Rett syndrome and age-matched healthy girls. Reduction of heart rate variability progresses with age and with the clinical stage of the syndrome. These results suggest the possible role of the progressive cardiac dysfunction in the sudden death associated with Rett syndrome. PMID- 11392525 TI - Rasmussen encephalitis associated with segmental vitiligo of the scalp: clinicopathologic report. AB - A 3-year-old boy with segmental vitiligo, poliosis, and alopecia over the right side of the scalp developed epilepsia partialis continua involving the left side of the body and progressive atrophy of the right cerebral hemisphere. There was a right ear dysacusia and a perilimbal vitiligo associated with an area of iris depigmentation in the right eye. Pleocytosis and hypergammaglobulin were detected in cerebrospinal fluid. Because medical treatment that included phenobarbital, phenytoin, carbamazepine, oxcarbazepine, benzodiazepines, corticosteroids, gamma globulin, and a ketogenic diet was ineffective, he underwent a right hemispherectomy. Neuropathologic examination showed a widespread scattered inflammatory process with numerous microglial nodules and perivascular lymphocytic cuffing associated with degenerative changes with severe neuronal loss, loosening of the neuropil, and microcystic changes with tissue collapse. The coexistence of vitiligo and possibly Vogt-Koyanagi-Harada syndrome in this child reinforces the autoimmune theory as the pathogenesis of Rasmussen syndrome. PMID- 11392526 TI - 31Phosphorus magnetic resonance spectroscopy in late-onset Tay-Sachs disease. AB - The late-onset form of GM2 gangliosidosis (Tay-Sachs disease) is an autosomal recessive disorder with progressive neurologic disease, mainly characterized by motor neuron and spinocerebellar dysfunction. The majority of patients are of Ashkenazi Jewish origin. 31Phosphorus magnetic resonance spectroscopy of the brain was performed to study the metabolic changes of a 16-year-old patient with late-onset Tay-Sachs disease who had a heterozygous Gly269-->Ser mutation in the hexosaminidase A encoding gene in compound heterozygosity with another, yet unidentified mutation. Severe changes in phosphorus metabolism with a decreased amount of phosphodiesters and membrane-bound phosphates were demonstrated, suggesting an activation of phosphodiesterases by accumulating gangliosides. The clinical findings were well related to the changes in spectroscopically determined metabolites. PMID- 11392527 TI - Clonidine treatment increases tics in patients with Tourette syndrome: case report. AB - Clonidine usually serves as a tic-suppressing agent in patients with Tourette syndrome. A case study is presented in which clonidine at subclinical concentrations led to an exacerbation of the tics and caused severe systemic heat sensations. Involvement of functional abnormalities of the hypothalamus in Tourette syndrome is hypothesized. PMID- 11392528 TI - Seckel's syndrome and malformations of cortical development: report of three new cases and review of the literature. AB - Seckel's syndrome is a rare form of primordial dwarfism, characterized by peculiar facial appearance. In the past, this condition was overdiagnosed, and most attention was given to the facial and skeletal features to define more precise diagnostic criteria. The presence of mental retardation and neurologic signs is one of the peculiar features of this syndrome, but only recently were rare cases of malformation of cortical development described, as documented by magnetic resonance imaging (MRI). Here, we present three new cases of Seckel's syndrome showing different malformations of cortical development (one gyral hypoplasia, one macrogyria and partial corpus callosum agenesis, and one bilateral opercular macrogyria). We hypothesize that the different types of clinical expression of our patients could be explained by different malformation of cortical development types. We think that MRI studies could be performed in malformative syndromes because of the possible correlations between type and extent of the lesion and the clinical picture of any individual case. PMID- 11392529 TI - Saccharomyces boulardii: potential adjunctive treatment for children with autism and diarrhea. PMID- 11392530 TI - The synthesis of amino-acid functionalized beta-carbolines as topoisomerase II inhibitors. AB - The synthesis and biological activity of amino acid functionalized beta-carboline derivatives, which are structurally related to azatoxin and the tryprostatins, are reported. These compounds were assayed for their growth inhibition properties in H520 and PC3 cell lines and were examined for their abilities to inhibit topoisomerase II-mediated DNA relaxation. PMID- 11392531 TI - Diaryl ether inhibitors of farnesyl-protein transferase. AB - Imidazolemethyl diaryl ethers are potent inhibitors of farnesyl-protein transferase. The SNAr displacement reaction used to prepare these diaryl ethers was amenable to rapid parallel synthesis of FPTase inhibitors. The use of a broad range of commercially available phenols quickly identified compounds which proved active in cells. PMID- 11392532 TI - Synthesis of polyamine derivatives having non-hypotensive Ca2 +-permeable AMPA receptor antagonist activity. AB - In order to obtain non-hypotensive and Ca2+-permeable AMPA receptor antagonists, we have synthesized a series of 1,4-bis(4-piperidinylmethyl)diaminobutanes. Compounds 13b, c, f had desirable properties. PMID- 11392533 TI - Phospho-Azatyrosine, a less effective protein-tyrosine phosphatase substrate than phosphotyrosine. AB - Azatyrosine (AzaTyr, 4) is a natural product isolated from Streptomyces chibanesis, whose structure is characterized by a nitrogen atom in the aryl ring of a tyrosyl residue. This seemingly minor modification to the tyrosyl residue results in profound physiological effects, as AzaTyr has been shown to promote permanent reversion of ras-dependent transformed cells to the normal phenotype in culture and to inhibit chemical induction of carcinogenesis in transgenic mice bearing oncogenic human ras. The mechanisms underlying these effects are not known, however ras-pathways involve an intricate balance between both protein tyrosine kinases (PTKs) and protein-tyrosine phosphatases (PTPs). The present study was undertaken to examine the general utility of AzaTyr as a structural motif for PTP inhibitor design by examining the phospho-azatyrosine (pAzaTyr) containing peptide Ac-Asp-Ala-Asp-Glu-pAzaTyr-Leu-amide (8) in a PTP1 enzyme system. Kinetic analysis indicated that 8 binds with a Km value of 210 microM and a catalytic turnover rate, kcat of 52 s(-1). This represents a greater than 50 fold reduction in binding affinity relative to the parent phosphotyrosine containing peptide, indicating that the aryl nitrogen adversely affects binding affinity. The much lower PTP affinity of the pAzaTyr-containing peptide reduces the potential utility of the AzaTyr pharmacophore for PTP inhibitor design. These results are discussed from the point of view that incorporation of AzaTyr residues into proteins could result in perturbation of protein-tyrosine phosphorylation,dephosphorylation cascades that control signal transduction processes, including ras-dependent pathways. PMID- 11392534 TI - Liver cell specific targeting of peptide nucleic acid oligomers. AB - Chimeric molecules consisting of peptide nucleic acid (PNA) and lactose have been synthesized to test the hypothesis that lactose moieties can promote cell specific uptake of PNAs. We find that lactose modified PNAs rapidly enter liver derived HepG2 cells while unmodified PNAs do not and that lactose modified PNAs can inhibit cellular telomerase. PMID- 11392535 TI - Synthesis and antifungal activities of novel 1,3-beta-D-glucan synthase inhibitors. Part 2. AB - Highly potent 1,3-beta-D-glucan synthase inhibitors, 7b, 10a, 10b and 12, have been identified by the chemical modification of the ornithine residue of a fungicidal macrocyclic lipopeptidolactone, RO-09-3655 (1), isolated from the cultured broth of Deuteromycotinia spp. These compounds showed stronger antifungal activity against systemic candidiasis as well as pulmonary aspergillosis in mice, and less hepatotoxicity as compared with 1. PMID- 11392536 TI - Synthesis and evaluation of a difluoromethylene analogue of sphingomyelin as an inhibitor of sphingomyelinase. AB - A sphingomyelin analogue 2, in which the long alkenyl chain and the phosphodiester moiety of sphingomyelin were replaced by a phenyl and an isosteric difluoromethylenephosphonic acid, was prepared to evaluate its inhibitory potency to sphingomyelinase. The analogue non-competitively inhibited the neutral sphingomyelinase in bovine brain microsomes with an IC50 of 400 microM. The compound had the ability to suppress tumor necrosis factor alpha-induced apoptosis of PC-12 neurons at a low concentration of 0.1 microM. PMID- 11392537 TI - Synthesis and evaluation of hexahydrochrysene and tetrahydrobenzofluorene ligands for the estrogen receptor. AB - To prepare novel estrogen receptor (ER) ligands, we have developed a facile approach to substituted hexahydrochrysene and tetrahydrobenzo[a]fluorene systems. Substituents, including basic side chains, were added to these systems, and their binding affinity to ERalpha and ERbeta, and in some cases their transcriptional activity were evaluated. PMID- 11392538 TI - Synthesis and biological evaluations of condensed pyridine and condensed pyrimidine-based HMG-CoA reductase inhibitors. AB - A series of 3,5-dihydroxyheptenoic acid derivatives containing pyrazolopyridine, isoxazolopyridine, thienopyridine, and pyrazolopyrimidine as a key scaffold was synthesized from condensed pyridine and condensed pyrimidine carboxylic acid esters by homologation, aldol condensation with ethyl acetoacetate dianion, and stereoselective reduction of the 5-hydroxyketone. Several compounds in the series were found to have potent HMG-CoA reductase inhibitory activities in vitro and marked cholesterol biosynthesis inhibitory activities in vivo. It has been shown that these scaffolds can be used as a suitable replacement for the hexahydronaphthalene ring present in naturally occurring HMG-CoA reductase inhibitors. PMID- 11392539 TI - Spirocyclic nonpeptide glycoprotein IIb-IIIa antagonists. Part 1: design of potent and specific 3,9-diazaspiro[5.5]undecanes. AB - The synthesis and biological activity of novel glycoprotein IIb-IIla antagonists containing the 3,9-diazaspiro[5.5]undecane nucleus are described. The potent activity of these compounds as platelet aggregation inhibitors demonstrates the utility of the spirocyclic structures as a central template for nonpeptide RGD mimics. PMID- 11392540 TI - Spirocyclic nonpeptide glycoprotein IIb-IIIa antagonists. Part 2: design of potent antagonists containing the 3-azaspiro[5.5]undecanes. AB - The synthesis and biological activity of novel glycoprotein IIb-IlIa anatagonists containing 3-azaspiro[5.5]undec-9-yl nucleus are described. The potent activity of these compounds as platelet aggregation inhibitors demonstrates the utility of the monoazaspirocyclic structure as central template for nonpeptide RGD mimics. PMID- 11392541 TI - Identification of a potent and selective oxytocin antagonist, from screening a fully encoded differential release combinatorial chemical library. AB - A library of 1,296 1,4-benzodiazepines was prepared on 160 microM Tentagel beads. Compounds are attached to the beads using orthogonally cleavable linkers. The library was first screened as pools of 30 beads where 50% of the material is released and screened. GW405212X, a selective oxytocin antagonist, was identified by picking single beads from active pools. PMID- 11392542 TI - Structure-activity relationship investigations of a potent and selective benzodiazepine oxytocin antagonist. AB - We have investigated the structure-activity relationships of the 1- and 3 substituents and replacements of the 5-phenyl group of GW405212X 1, a potent selective oxytocin antagonist. The effect of these modifications on oxytocin binding antagonism and on pharmacokinetic parameters is reported. PMID- 11392543 TI - Structure-Activity study of retinoid agonists bearing substituted dicarba-closo dodecaborane. Relation between retinoidal activity and conformation of two aromatic nuclei. AB - We have investigated the structure activity relationships of the potent retinoid agonist, 4-[4-(2-propyl-1,2-dicarba-closo-dodecaboran-l-yl)phenylamino]benzoic acid (BR403), which we have previously reported. Substitution of a methyl group on the aromatic nucleus or a methyl group on the nitrogen atom, or replacement of the amino group with ether, methylene, carboxyl or 1,1-ethylene greatly decreased the activity. The relatively planar conformation at the phenyl-N-phenyl moiety seems to play a critical role in the appearance of the biological activity. PMID- 11392544 TI - Pharmacological evaluation of a diarylmethylene-piperidine derivative: a new potent atypical antipsychotic? AB - A new diaryl-methylene piperidine derivative, 2, displayed an atypical antipsychotic profile both in vitro and in vivo. The main pharmacological characteristics of this compound appears to reside in a more potent antagonism of the 5-HT2 serotonergic receptor than of the D2 dopaminergic receptor. This confirms that molecules displaying a D2/5-HT2 binding ratio < 1 possess clozapine like antipsychotic activity. PMID- 11392545 TI - A new structural class of selective and non-covalent inhibitors of the chymotrypsin-like activity of the 20S proteasome. AB - We describe the identification and in vitro characterization of a series of 2 aminobenzylstatine derivatives that inhibit non-covalently the chymotrypsin-like activity of the 20S proteasome. Our initial SAR data demonstrate that the 2 aminobenzylstatine core structure can effectively serve as the basis for designing potent, selective and non-covalent inhibitors of the chymotrypsin-like activity of the 20S proteasome. PMID- 11392546 TI - Modeling of the binding mode of a non-covalent inhibitor of the 20S proteasome. Application to structure-based analogue design. AB - The 2-aminobenzvlstatine derivative I is a 20S proteasome inhibitor of a novel chemical type identified by high throughput screening. The compound specifically inhibits the chymotrypsin-like catalytic activity of the human proteasome with an IC50 value in the micromolar range. Using the crystal structure of the yeast proteasome, we modeled the structure of the human proteasome in complex with 1. As one of the first applications of the model in our oncology programme targeting the proteasome, we designed an analogue of the inhibitor having enhanced stacking/hydrophobic interactions with the enzyme. One order of magnitude in inhibitory potency was gained. PMID- 11392547 TI - Thiazole analogues of the NSAID indomethacin as selective COX-2 inhibitors. AB - The carboxyl group of the NSAID indomethacin was replaced with a variety of substituted thiazoles to obtain a series of potent, selective inhibitors of COX 2. Additional substitutions were made at the 1-position and 5-position of the indole of indomethacin. PMID- 11392548 TI - Design, synthesis and enzymatic activity of highly selective human mitochondrial thymidine kinase inhibitors. AB - Highly selective arabinofuranosyl nucleosides, which inhibit the mitochondrial thymidine kinase (TK-2) without affecting the closely related herpes simplex virus type 1 thymidine kinase (HSV-1 TK), varicella-zoster virus thymidine kinase (VZV-TK), cytosolic thymidine kinase (TK-1) or the multifunctional Drosophila melanogaster deoxyribonucleoside kinase (Dm-dNK), have been obtained. SAR studies indicate a close relation between the length of the substituent at the 2' position of the arabinofuranosyl moiety and the inhibitory activity. PMID- 11392549 TI - Ring-Constrained (N)-methanocarba nucleosides as adenosine receptor agonists: independent 5'-uronamide and 2'-deoxy modifications. AB - Novel methanocarba adenosine analogues, having the pseudo-ribose northern (N) conformation preferred at adenosine receptors (ARs), were synthesized and tested in binding assays. The 5'-uronamide modification preserved [N6-(3-iodobenzyl)] or enhanced (N6-methyl) affinity at A3ARs, while the 2'-deoxy modification reduced affinity and efficacy in a functional assay. PMID- 11392550 TI - Powerful probes for glycosidases: novel, fluorescently tagged glycosidase inhibitors. AB - Amino-1,2,5-trideoxy-2,5-imino-D-mannitol was fluorescently tagged by reaction with dansyl chloride at N-1 or by attachment of a dansyl amide bearing spacer to this centre. Compounds obtained are highly potent inhibitors of beta-glucosidase exhibiting Ki values in the single figure nanomolar range. The 1-N-dansyl substituted inhibitor was successfully exploited for binding studies with beta glucosidase from Agrobacterium sp. employing fluorescence spectrometric methods. PMID- 11392551 TI - A new method for chemoselective conjugation of unprotected peptides to dauno- and doxorubicin. AB - A new approach for chemoselective ligation of peptides to dauno- and doxorubicin through an oxime bond is presented. The method does not require protecting groups on the peptide moiety. PMID- 11392552 TI - Anodic oxidation of ifosfamide and cyclophosphamide: a biomimetic metabolism model of the oxazaphosphorinane anticancer drugs. AB - The electrochemical oxidation of anticancer drugs ifosfamide and cyclophosphamide produced in high yield methoxylated analogues of the key hydroxy-metabolites of these oxazaphosphorine prodrugs. The cytotoxicity of these compounds was evaluated, and found to be as high as the hydroxy-metabolite. PMID- 11392553 TI - Dynamics of actin and alpha-actinin in nascent myofibrils and stress fibers. AB - Actin labeled with fluorescein isothiocyanate (FITC) and alpha-actinin labeled with rhodamine (rh) were co-injected into chick embryonic cardiac myocytes and fibroblasts. In cardiomyocytes, FITC-actin was distributed in nonstriated lines, linearly arranged punctate structures with short intervals, and cross-striated bands with regular sarcomeric intervals. rh-alpha-Actinin was seen to be distributed in the same pattern in the former two portions, and in the center of each striation in the latter portion. Photobleaching of structures incorporated with these fluorescent analogs revealed that the fluorescent recovery rate of actin decreased in the order of nonstriated > punctated > striated portions, while that of alpha-actinin was low and stable at all portions. During the transition phase from punctate to regular sarcomere structures of these proteins, short spaced alpha-actinin dots adjoined each other and aligned with Z bands of neighboring myofibrils. It appears that both the difference in exchangeability between actin and alpha-actinin molecules and the movement of alpha-actinin dots during this phase of myofibrillogenesis are related to sarcomere lengthening and I-Z-I brush formation; adjoining dots of low-exchangeable alpha-actinin may provide favorable situations for exchangeable actin molecules in filaments to elongate and/or rearrange. In fibroblasts, both FITC-actin and rh-alpha-actinin formed nonstriated lines. In these cells, exchangeabilities of both proteins were high and similar in rate. This seems to indicate that stress fibers are constantly exchanging their components for motile and other vital functions of these cells. The high exchangeabilities of both proteins in stress fibers showthat these fibers are clearly different from nonstriated, stress-fiber like structures of nascent myofibrils. PMID- 11392554 TI - Dynamics of stimulation-induced muscle adaptation: insights from varying the duty cycle. AB - We sought to gain insight into the dynamics of the signalling process that initiates adaptive change in mammalian skeletal muscles in response to chronic neuromuscular stimulation. Programmable miniature stimulators were implanted into rabbits and used to impose one of the following patterns on the dorsiflexors of one ankle: 10 Hz delivered in equal on/off periods of 30 s, 30 min, or 12 h (all equivalent in terms of aggregate impulse activity to continuous 5 Hz). Two further groups received continuous stimulation at 5 Hz or 10 Hz. In every case the stimulation pattern was maintained continuously for 6 weeks. Tibialis anterior muscles stimulated intermittently with equal on/off periods of 30 s, 30 min and 12 h had contractile characteristics that were significantly slower than the contralateral, unstimulated muscles but did not differ from those of muscles stimulated continuously at 5 Hz. Muscles stimulated continuously at 10 Hz were significantly slower than either contralateral muscles or muscles stimulated with any of the other patterns. Corresponding changes were seen in myosin heavy chain isoform composition. The fatigue index, defined as the fraction of tension remaining after 5 min of a standard fatigue test, was 0.4 for muscles in the contralateral group but equal to or greater than 0.85 for muscles of all the stimulated groups. These results were interpreted with the help of a simple model of the growth and decay of a putative signalling substance based on first order kinetics. The model suggests a rate constant for the accumulation of the signalling substance that is greater than 30 h(-1), and a rate constant for its removal that is greater than 50 h(-1). PMID- 11392555 TI - Factors contributing to troponin exchange in myofibrils and in solution. AB - The troponin complex in a muscle fiber can be replaced with exogenous troponin by using a gentle exchange procedure in which the actin-tropomyosin complex is never devoid of a full complement of troponin (Brenner et al. (1999) Biophys J 77: 2677 2691). The mechanism of this exchange process and the factors that influence this exchange are poorly understood. In this study, the exchange process has now been examined in myofibrils and in solution. In myofibrils under rigor conditions, troponin exchange occurred preferentially in the region of overlap between actin and myosin when the free Ca2+ concentration was low. At higher concentrations of Ca2+, the exchange occurred uniformly along the actin. Ca2+ also accelerated troponin exchange in solution but the effect of S1 could not be confirmed in solution experiments. The rate of exchange in solution was insensitive to moderate changes in pH or ionic strength. Increasing the temperature resulted in a two-fold increase in rate with each 10 degrees C increase in temperature. A sequential two step model of troponin binding to actin-tropomyosin could simulate the observed association and dissociation transients. In the absence of Ca2+ or rigor S1, the following rate constants could describe the binding process: k1 = 7.12 microM(-1) s(-1), k(-1) = 0.65 s(-1), k2 = 0.07 s(-1), k(-2) = 0.0014 s(-1). The slow rate of detachment of troponin from actin (k(-2)) limits the rate of exchange in solution and most likely contributes to the slow rate of exchange in fibers. PMID- 11392556 TI - Hydrogen peroxide modulates Ca2+-activation of single permeabilized fibres from fast- and slow-twitch skeletal muscles of rats. AB - We examined the effects of redox modulation on single membrane-permeabilized fibre segments from the fast-twitch extensor digitorum longus (EDL) and slow twitch soleus muscles of adult rats to determine whether the contractile apparatus was the redox target responsible for the increased contractility of muscles exposed to low concentrations of H2O2. The effects of H2O2 on maximum Ca2+-activated force were dose-dependent with 30 min exposure to 5 mM H2O2 causing a progressive decrease by 22+/-3 and 13+/-2% in soleus and EDL permeabilized muscle fibres, respectively. Lower concentrations of exogenous H2O2 (100 microM and 1 mM) had no effect on maximum Ca2+-activated force. Subsequent exposure to the reductant dithiothreitol (DTT, 10 mM, 10 min) fully reversed the H2O2-induced depression of force in EDL, but not in soleus muscle fibres. Incubation with DTT alone for 10 min did not alter Ca2+-activated force in either soleus or EDL muscle fibres. The sensitivity of the contractile filaments to Ca2+ (pCa50) was not altered by exposure to any concentration of exogenous H2O2. However, all concentrations of H2O2 diminished the Hill coefficient in permeabilized fibres from the EDL muscle, indicating that the cooperativity of Ca2+ binding to troponin is altered. H2O2 (5 mM) did not affect rigor force, which indicates that the number of crossbridges participating in contraction was not reduced. In conclusion, H2O2 may reduce the maximum Ca2+ activated force production in skinned muscle fibres by decreasing the force per crossbridge. PMID- 11392557 TI - Fibre types and myosin heavy chain expression in the ocular medial rectus muscle of the adult rat. AB - Myosin heavy chain (MHC) expression was determined immunohistochemically in individual muscle fibre types characterised by activities of ATPase and the key oxidative and glycolytic enzymes in rat ocular medial rectus (MR) muscles. In the global layer (GL), glycolytic activity of muscle fibres was higher and oxidative activity lower, than in the orbital layer (OL). Muscle fibres in the former displayed rosette-like organisation with a slow fibre surrounded by several fast fibres, which expressed either MHCIIa or MHCIIb, but many co-expressed both isoforms. In the OL some slow fibres co-expressed MHCIIa. Extraocular MHC isoform (MHCeom) could not be determined immunohistochemically and no pure MHCIIx/d containing fibres were found, suggesting that these isoforms, demonstrated electrophoretically, are co-expressed with others. Slow muscle fibres in both layers co-expressed MHCbeta slow, MHCalpha cardiac and MHC-slow tonic. Neonatal isoform (MHCneo) was co-expressed in several fast and slow muscle fibres in the orbital, but not global layer. Slow fibres in the GL displayed very low oxidative activity. Electrophoretic analysis of ocular MR muscle homogenates revealed that about 50% of total MHC was MHCIIb, MHCeom was quite prominent (25%), and MHCIIa, MHCIIx/d and MHCI contributed each about 8%. MHCneo, MHCslow tonic and MHCalpha cardiac could not be identified as separate bands. PMID- 11392558 TI - Protein and mRNA levels of the myosin heavy chain isoforms Ibeta, IIa, IIx and IIb in type I and type II fibre-predominant rat skeletal muscles in response to chronic alcohol feeding. AB - Alcoholic myopathy occurs in between one and two-thirds of all alcohol misusers and is thus one of the most prevalent muscle disorders (2000 cases per 100,000 population). It is characterised by myalgia, muscle weakness and loss of lean tissue mass. Histological features include a reduction in the diameter of Type II muscle fibres, particularly the IIb fibre subset. In contrast, Type I fibres are relatively protected. It is possible that the myopathy is due to perturbations in myosin protein and mRNA expression. To test this hypothesis, we fed rats a liquid diet containing 35% of calories as ethanol. Control rats were pair-fed identical amounts of the same diet in which ethanol was replaced by isocaloric glucose. At the end of 6 weeks, total myofibrillary proteins and myosin heavy chain (MyoHC) Ibeta, IIa, IIx and IIb protein and mRNA were analysed in the plantaris (Type II fibre-predominant) and soleus (Type I fibre-predominant) muscles. The data showed that there were significant reductions in the total myofibrillary protein content in the plantaris of ethanol fed rats compared to pair-fed controls (P < 0.05). These changes in the plantaris were accompanied by reductions in total myosin (P < 0.025), as a consequence of specific reductions in the Ibeta, (P < 0.01), IIx (P < 0.05) and IIb (P < 0.05) protein isoforms. The mRNA levels of Ibeta were significantly reduced in the plantaris (P < 0.05). However, mRNA levels of IIa, IIx and IIb in the plantaris were not significantly affected by alcohol feeding. Other changes in the plantaris included significant reductions in desmin (P < 0.01), actin (P < 0.025), and troponin-I (P < 0.05) compared to pair-fed controls. In the soleus, the only significant changes related to a fall in Ibeta mRNA levels and a decline in troponin-C content. We conclude that in the rat, alcoholic myopathy is a feature of Type II fibre rich muscles and is accompanied by multiple protein changes. The decline in specific myosin protein levels, such as IIx and IIb in the absence of corresponding reductions in their mRNAs, is probably due to altered proteolysis or more likely reductions in translational efficiencies, rather than changes in transcription. PMID- 11392559 TI - Remodeling of the actin cytoskeleton in the contracting A7r5 smooth muscle cell. AB - It has been proposed that the reorganization of components of the actin cytomatrix could contribute to force development and the low energy cost of sustained contraction in contractile cells which lack a structured sarcomere (A.S. Battistella-Patterson, S. Wang and G.L. Wright (1997) Can J Physiol Pharmacol 75: 1287-1299). However, there has been no direct evidence of an apropos actin reorganization specifically linked to the contractile response in cells of this type. Remodeling of the alpha- and beta-actin domains was studied in A7r5 smooth muscle cells during phorbol 12,13 dibutyrate (PDB)-induced contraction using immunohistologic staining and beta-actin-green fluorescent protein (beta-actin-GFP) fusion protein expression. Cell stained with phalloidin as well as cells expressing beta-actin-GFP showed densely packed actin stress cables, arranged in parallel and extending across the cell body. PDB caused approximately 85% of cells to contract with evidence of forcible detachment from peripheral adhesion sites seen in many cells. The contraction of the cell body was not uniform but occurred along a principal axis parallel to the system of densely packed beta-actin cables. During the interval of contraction, the beta actin cables shortened without evidence of disassembly or new cable formation. The use of cytochalasin to inhibit actin polymerization resulted in the dissolution of the actin cables at the central region of the cell and caused the elongation of precontracted cells. In unstimulated cells, alpha-actin formed cables similar in arrangement to the cell spanning beta-actin cables. Within a short interval after PDB addition; however, the majority of alpha-actin cables disassembled and reformed into intensely fluorescing column-like structures extending vertically from the cell base at the center of clusters of alpha-actin filaments. The alpha-actin columns of contracting cells showed strong colocalization of alpha-actinin suggesting they could be structurally analogous to the dense bodies of highly differentiated smooth muscle cells. The results indicate that the alpha- and beta-actin domains of A7r5 cells undergo a highly structured reorganization during PDB-induced contraction. The extent and nature of this restructuring suggest that remodeling could play a role in contractile function. PMID- 11392560 TI - The executive functions and self-regulation: an evolutionary neuropsychological perspective. AB - Neuropsychology has customarily taken a molecular and myopic view of executive functioning, concentrating largely on those proximal processes of which it may be comprised. Although commendable as a starting point, such an approach can never answer the question, "Why executive functioning?" The present paper encourages neuropsychologists to contemplate the longer-term, functional nature of the executive functions (EFs), using an evolutionary perspective. For purely illustrative purposes, a previously developed model of the EFs is briefly presented and is then examined from an evolutionary perspective. That model views the EFs as forms of behavior-to-the-self that evolved from overt (public) to covert (private) responses as a means of self-regulation. That was necessary given the interpersonal competition that arises within this group-living species. The EFs serve to shift the control of behavior from the immediate context, social others, and the temporal now to self-regulation by internal representations regarding the hypothetical social future. The EFs seem to meet the requirements of a biological adaptation, being an improbable complex design for a purpose that exists universally in humans. Discovering the adaptive problems that the EFs evolved to solve offers an invaluable research agenda for neuropsychology lest that agenda be resolved first by other scientific disciplines. Some adaptive problems that the EFs may have evolved to solve are then considered, among them being social exchange (reciprocal altruism or selfish cooperation), imitation and vicarious learning as types of experiential theft, mimetic skill (private behavioral rehearsal) and gestural communication, and social self-defense against such theft and interpersonal manipulation. Although clearly speculative at the moment, these proposals demonstrate the merit of considering the larger adaptive problems that the EFs evolved to solve. Taking the evolutionary stance toward the EFs would achieve not only greater insight into their nature, but also into their assessment and into those larger adaptive capacities that may be diminished through injury or developmental impairment toward that system. PMID- 11392561 TI - Quantitative electroencephalography (QEEG) and neuropsychological syndrome analysis. AB - The ideographic, syndrome analysis and the nomothetic, standardized test battery approaches to neuropsychological assessment are compared and contrasted within the context of advances in noninvasive technology readily available for use within the examiner's office. By demonstrating the relative strengths and benefits of syndrome analysis, it is suggested that this approach provides a thorough and efficient method of neuropsychological assessment. Subsequently, the utility of an a priori hypothesis testing process approach as a critical technique in syndrome analysis will be supported. It will be proposed that QEEG procedures provide a useful method for further substantiating conclusions generated from a syndrome analysis approach to neuropsychological assessment. Two cases are described demonstrating the utility and flexibility of the QEEG as a confirmatory test of localization following syndrome analysis. In summary, the contributions that neuropsychologists make to the understanding of brain-behavior relationships may be strengthened by combining neuropsychological and neurophysiological assessment methods. PMID- 11392562 TI - Sources of heterogeneity in schizophrenia: the role of neuropsychological functioning. AB - Although schizophrenia is often characterized as a heterogeneous disorder, efforts to validate stable and meaningful subtypes have met with limited success. Thus, the issue of whether schizophrenia reflects a continuum of severity or a number of discrete subtypes remains controversial. This review evaluates efforts to establish subtypes based upon a model that includes causes, characteristics, and course and outcomes of heterogeneity. Emphasis is placed on empirical classification studies utilizing cognitive tests or symptom rating scales, sometimes in conjunction with neuroimaging procedures. Results of recent cluster analytic studies are reviewed that produced evidence of four or five clusters, varying in level and pattern of performance. Although this research typically generated meaningful subtypes, it was often the case that there was little correspondence between subtyping systems based upon cognitive function and those based upon symptom profile. It was concluded that there may be different mechanisms for producing cognitive and symptomatic heterogeneity, and that diversity in presentations of schizophrenia reflects a combination of continuities in severity of the disorder with a number of meaningful and stable subtypes. PMID- 11392563 TI - Extracellular enzymatic activity in 11 Cryptococcus species. AB - The extracellular enzymatic activity of 36 strains of yeast belonging to 11 species of the genus Cryptococcus, has been investigated, using the API-ZYM (BioMerieux, France) commercial system, with the objective of determining the differences in the enzymatic profiles of the various species. The strains studied were: 9 of C. neoformans, 7 of C. albidus, 6 of C. laurentii, 5 of C. uniguttulatus, 3 of C. humicolus, and 1 each of C. ater, C. curvatus, C. dimennae, C. hungaricus, C. infirmo-miniatus and C. magnus. All the strains showed enzymatic activity with positivity to Phosphatase alkaline, Esterase lipase C8, Leucine arylamidase, Phosphatase acid and Naphthol-AS-BI phosphohydrolase, and negativity to Lipase C14, Trypsin, Chemotrypsin, beta galactosidase, beta-glucuronidase and alpha-manosidase. Variable enzymatic activity was shown to Esterase C4, Valine arylamidase, Cystine arylamidase, alpha galactosidase, alpha-glucosidase, beta-glucosidase, N-acetyl-beta-glucosaminidase and alpha-fucosidase. This allowed 11 separate enzymatic patterns to be established. The species C. neoformans and C. laurentii each presented two distinct patterns; C. uniguttulatus, C. hungaricus and C. magnus shared the same pattern; C. albidus, C. ater, C. curvatus, C. dimennae, C. humicolus and C. infirmo-miniatus presented an individual enzymatic pattern. The results obtained suggest that the API-ZYM system could be useful for the identification of species of the genus Cryptococcus and for the differentiation of the enzymotypes for epidemiological purposes. PMID- 11392564 TI - A review of soybean (Glycine max) seed, pod, and flower mycofloras in North America, with methods and a key for identification of selected fungi. AB - A review of the fungi associated with soybean seeds, pods, or flowers was conducted in North America. Species of Deuteromycetes are the most common fungi in each of the soybean flower organs followed by the Ascomycetes and Phycomycetes which comprise about one-fourth of the total mycoflora. Eighty genera and about 135 or more species occur in seeds, pods, or flowers. With regard to numbers of taxa from separate mycofloras, 63 genera and about 108 or more species occur in seeds, 65 genera and about 88 or more species occur in pods, and 36 genera and approximately 47 or more species occur in flowers. Most of the fungi which occur in flowers can be cultured from pods, and the majority of those fungi occur in seeds. Methods for and a key are provided for the identification of the 30 most important selected fungi. PMID- 11392565 TI - Pythium spp. present in irrigation water in the poniente region of Almeria (south eastern Spain). AB - Pythium catenulatum, P. diclinum and P. paroecandrum, as well as asexual forms with filamentous sporangia were isolated from irrigation water in the Poniente region of Almeria (south-eastern Spain). The taxonomic and morphological details of these fungal isolates are described. P. catenulatum and P. diclinum are new reports to Spain. PMID- 11392566 TI - Fungal occurrence, disease incidence and severity, and yield of maize symptomatic for seedling disease in Mississippi. AB - A study was conducted in Mississippi from 1995 to 1997 comparing soil rhizosphere fungi isolated from Pioneer 3167 hybrid maize (Zea mays L.) planted on Brooksville silty clay and Memphis silt loam soils. Maize seedlings were collected over four sampling dates from conventional and no-tillage plots. Eleven fungal genera consisting of nineteen species were isolated from these plants; Trichoderma spp. were most frequently isolated, followed by Fusarium spp. The highest disease incidence occurred in tilled plots of the latest planting date on Brooksville silty clay when samples were collected 17 days after planting. Root disease was most severe in 1996 from seedlings planted on the last planting date in tilled plots sampled 17 days after planting. Yields were significantly (P < or = 0.05) higher on Brooksville silty clay soil than on Memphis silt loam in both 1995 and 1996. Yields were highest from no-tillage plots and from maize planted on the earliest date. There was a significant correlation between incidence of root infection and disease severity. There was no correlation between the incidence of root infection and yield or between disease severity and yield at either location. PMID- 11392567 TI - Survey of yeast mastitis in dairy herds of small-type farms in the Lublin region, Poland. AB - The purpose of this study was to isolate fungi from the quarter milk of cow udders in 9 dairy farms. The samples were collected between May 1996 and April 2000 in the Lublin region. Six hundred and four milk samples collected from the quarters of 172 cows with clinical and subclinical mastitis were examined. Milk samples were plated as soon as possible on blood agar (BA), Mac Conkey agar, aesculin-tallium acetate-crystal violet blood agar, and Sabouraud agar with chloramphenicol and gentamicin. Fifty eight (9.6%) of the samples were positive for fungi. All of the fungal isolates were yeasts of the genera Candida, Rhodotorula and Trichosporon. We also isolated Streptococcus agalactiae (4.9%), Str. spp.(6.4%), Staphylococcus aureus (10.4%), coagulase-negative staphylococci CNS (36.6%), Escherichia coli (3.5%), other microorganisms (2.6%) and no growth (25.8%). PMID- 11392568 TI - Onychomycosis in Tehran, Iran: prevailing fungi and treatment with itraconazole. AB - A total of 187 Patients with suspected onychomycosis were examined for causative fungal agents between 1996 and 1997. Laboratory examination confirmed onychomycosis in 115 patients, of which 97 cases were presented with positive microscopic and cultural examinations, and they were selected for itraconazole pulse therapy. From an etiological point of view, 48.4% of the nail infections, mainly toenail infections, were caused by dermatophytes, 43.3% were infected with Candida spp, specially infected fingernails, and 8.2% by non-dermatophytic molds. Trichophyton mentagrophytes var. interdigital and T. violaceum were the most prevalent species. Candida albicans and C. parapsilosis were the predominant species of the Genus Candida. Scopolariopsis brevicaulis was the most common non dermatophyte molds observed. Female affected more frequently than male and in both sexes, those who were 30-49 years old, more infected. Toenails were affected more frequently than fingernails. In this study, itraconazole pulse therapy (400 mg daily) gave during the first week of per month for 3 months. The study included 51 patients with toenail onychomychosis (group 1) and 46 patients with fingernail infections (group 2). Patients were followed up for 9 months after the last treatment. Clinical response rates were 83% in the group 1, 95% in the group 2 at month 12; the corresponding mycological cure rates were 71 and 87%, respectively. PMID- 11392569 TI - Exudation of plasma and production of thromboxane in human bronchi after local bradykinin challenge. AB - Plasma exudation has been suggested to be an important component of the inflammatory response in asthma. Bradykinin elicits many of the features of asthma, including bronchoconstriction, cough, plasma exudation and mucus secretion. In an attempt to quantify local plasma exudation, we have employed a novel low-trauma technique with the aim of challenging and lavaging a central part of the bronchial tree, by selecting a medium sized bronchus. A fibreoptic bronchoscopy was performed in non-smoking healthy volunteers. The instrument was placed proximally in the right upper lobe bronchus. A plastic catheter, equipped with an inflatable latex balloon, was inflated with air (2-4 cmH2O). A solution (100 microl of either two different concentrations of bradykinin: 0.09 and 0.9 mg ml(-1) or normal saline) was instilled through the catheter and distal to the balloon. Eight minutes later a lavage procedure with 10 ml of saline was performed through the catheter. The procedure was then repeated twice, with the other solutions, but from the lingular and middle lobe bronchi. All solutions were given in a blinded fashion, and two different studies were performed. Lavage concentrations of albumin and IgG were quantified as measurements of plasma exudation. In our first study we found that bradykinin challenge significantly increased concentrations of albumin and IgG. In study two, there was no numeric increase in plasma proteins after local bradykinin challenge, but the concentration of thromboxane was significantly increased in lavages from bradykinin-challenged bronchi. Thus, local bronchial administration of bradykinin has the capacity to induce exudation of large plasma macromolecules into the bronchial lumen, as well as local thromboxane production. PMID- 11392570 TI - Measuring asthma control in group studies: do we need airway calibre and rescue beta2-agonist use? AB - Collection of airway calibre and beta2-agonist data in large clinical trials and epidemiological surveys is sometimes difficult and may be an inefficient use of resources. The aim of this study was to determine whether the omission of the forced expiratory volume in 1 sec (FEV1) and beta2-agonist questions from the seven-item Asthma Control Questionnaire (ACQ) alters its measurement properties and validity. In an observational study, 50 adults with symptomatic asthma attended the clinic at 0, 1, 5 and 9 weeks to complete the ACQ and other measures of asthma status. All patients completed the study and provided complete data sets. Omission of the FEV1 and beta2-agonist questions from the ACQ made minimal difference to the reliability, responsiveness, and both cross-sectional and longitudinal validity of the instrument. Omission of the FEV1 question significantly lowered the summary score (P<0.001) but omission of the beta2 agonist question did not alter it (P>0.05). In group studies, both the FEV1 and beta2-agonist questions may be omitted from the ACQ without changing the validity or the measurement properties of the instrument. Lowering of the summary score by the omission of the FEV1 question means that data from this abbreviated form cannot be combined with or compared to data collected using the full questionnaire. PMID- 11392571 TI - Ex-vivo product performance of Diskus and Turbuhaler inhalers using inhalation profiles from patients with severe chronic obstructive pulmonary disease. AB - Dosing performance of dry powder inhalers is dependent on patient's inspiratory effort. This study compares the inhalation profiles generated by patients with severe obstructive lung disease using Diskus and Turbuhaler inhalers. The patient profiles are subsequently used to determine the dosing performance of fluticasone propionate Diskus and budesonide Turbuhaler inhalers. Inhalation profiles were recorded in COPD patients (FEV1 < or = 30% predicted) as they inhaled with maximal effort through the inhalers. The profiles were used in an inhalation simulator to assess the dosing performance by measuring the total emitted dose and the fine particle mass for each inhaler type. Peak inspiratory flow was significantly higher through the Diskus (mean 82.31 min(-1)) compared with Turbuhaler (mean 53.51 min(-1), difference = 28.8 l min(-1); P < 0.0001). In addition, in direct comparison of the two devices. the Diskus was shown to deliver a more consistent dose irrespective of flow than the Turbuhaler in this patient population. These findings may be of importance in optimising selection of devices for patients with severe airway obstruction. PMID- 11392572 TI - Is continuous transcutaneous monitoring of PCO2 (TcPCO2) over 8 h reliable in adults? AB - Monitoring of non-invasive ventilation (NIV) in a non-intensive care unit (non ICU) setting requires a method of evaluating nocturnal PaCO2, such as transcutaneous CO2 monitoring (TcPCO2). However, changing the probe site after 4 h and recalibrating (as recommended) is time-consuming and impractical. Continuous (8-h) TcPCO2 monitoring at a lower electrode temperature (43 degrees C) in this setting has never been formally studied. Patients under intermittent NIV were studied (n = 28, aged 69 +/- 9 years, PaO2: 71 +/- 13 mmHg, PaCO2: 49 +/ 9 mmHg). After calibration and stabilization of TcPCO2 (Radiometer Tina TCM3 capnograph), arterial blood gases (ABG) were measured and compared with transcutaneous readings. In 10 patients who underwent continuous 8-h TcPCO2 recording, ABGs were also measured after 4 and 8 h. The correlation between TcPCO2 and PaCO2 was highly significant (r2 = 0.92, P<0.0001). Mean (TcPCO2 PaCO2) gradient (bias) was: -2.8 +/- 3.8 mmHg; limits of agreement were: (-10.4; +4.8 mmHg). TcPCO2-PaCO2 gradient was lowest (i.e. within-bias +/- 2 mmHg) between 40 and 54 mmHg, increasing below and above these values. Over 8 h, no significant drift of the TcPCO2 signal occurred (ANOVA). No discomfort or skin lesion was noted. In conclusion, with an electrode temperature of 43 degrees C, 8 h continuous monitoring of TcPCO2 was well tolerated, without any local side effects or significant drift of TcPCO2 signal; when compared to previous reports, lowering the electrode temperature did not decrease performance for CO2 monitoring. PMID- 11392573 TI - How accurate are pulse oximeters in patients with acute exacerbations of chronic obstructive airways disease? AB - The aim of this study was to determine the extent of correlation and agreement between arterial oxygen saturation and oxygen saturation as recorded by transcutaneous pulse oximetry, with a view to identifying whether pulse oximetry can be used as an alternative to arterial values in the clinical management of patients with acute exacerbations of chronic obstructive airways disease (COAD) in the emergency department. It also aims to determine whether there is a cut-off level of oxygen saturation by pulse oximetry that can screen for significant systemic hypoxia in this group. This prospective study of patients with acute exacerbations of COAD who were deemed by their treating doctor to require an arterial blood gas analysis to determine their ventilatory status, compared arterial oxygen saturation with simultaneously recorded oxygen saturation measured by transcutaneous pulse oximetry. Data were analysed using Pearson correlation, bias plot (Bland-Altman) methods for agreement and the receiver operator characteristic (ROC) curve method for determination of a screening cut off. Sixty-four sample-pairs were analysed for this study. Nine (14%) had significant hypoxia (arterial PO2 less than 60 mmHg). The correlation coefficient was 0.91. The bias (Bland-Altman) plot shows a constant bias of -0.758% and only fair agreement, with 95% limits for agreement of -8.2 to + 6.7%. With respect to the ROC curve analysis, the 'best' cut-off for detection of hypoxia was at oxygen saturation by pulse oximetry of 92% (sensitivity 100%, specificity 86%). In conclusion, there is not sufficient agreement for oxygen saturation measured by pulse oximetry to replace analysis of an arterial blood gas sample in the clinical evaluation of oxygenation in emergency patients with COAD. However, oxygen saturation by pulse oximetry may be an effective screening test for systemic hypoxia, with the screening cut-off of 92% having sensitivity for the detection of systemic hypoxia of 100% with specificity of 86%. PMID- 11392574 TI - Allergic bronchopulmonary mycosis in patients with asthma: period prevalence at a university hospital in Saudi Arabia. AB - Allergic bronchopulmonary mycosis (ABPM) is a known complication of asthma and can result in progressive lung damage, respiratory failure and death. Asthma is a common disease in Saudi Arabia and until now the prevalence of ABPM has not been investigated. The aim of this study was to estimate the period prevalence of ABPM due to Aspergillus and Candida in patients with asthma. The setting was an outpatient pulmonary clinic at a university hospital in the central region of Saudi Arabia. Two hundred and sixty-four consecutive patients with asthma (150 or 57% females) were evaluated. All patients were screened for ABPM with skin prick test (SPT) using a panel of fungal antigens. Those with positive skin reactions had further clinical, immunological, respiratory and radiological assessment. ABPM was diagnosed by the presence of a minimum of five of the major criteria suggested by Rosenberg in 1977. Of the 264 patients, 62 (23%) had a positive SPT for at least one fungal allergen, of whom 44 (71%) were females (P=0.01). Seven patients (six females) were diagnosed with ABPM due to Aspergillus and (or) Candida species. Therefore, we estimate the period prevalence of ABPM to be 2.7% (95% confidence interval 1.3-5.4%). A. niger was the commonest fungal species isolated in our group. In conclusion, ABPM is not uncommon in Saudi Arabia and females seem to be more at risk. Because asthma is common, physicians need to have high index of suspicion for this disease and pursue the diagnosis with the appropriate tests. PMID- 11392575 TI - Recurrent flu-like illness with migrating pulmonary infiltrates of unknown aetiology. AB - Migrating pulmonary infiltrates present a difficult diagnostic and therapeutic challenge. We report on eight patients (mean age 51 years, range 32-78 years, with a prolonged history of migrating pulmonary infiltrates of unknown aetiology despite a very elaborate search for infectious causes, hypersensitivity pneumonitis or inhalation fever due to occupational or domestic exposure to fungi, or to other environmental causes, and for humoral or cellular immunological incompetence. These patients (one male, seven females) presented with recurrent episodes (mean 6, range 2-13) of a flu-like illness, often with cough, wheezing and pleuritic chest pain, but without systemic involvement. Previous medical histories were unremarkable. There was no relation with smoking habits, occupation, drug use or other possible exposures. Biochemical data were non-specific. There was no peripheral nor pulmonary eosinophilia; total IgE was normal, with negative RASTs and precipitins to a variety of antigens. Cultures and serological tests for bacteria, viruses, fungi, etc were non-contributory. Chest X-ray and computed tomography (CT) scan showed bilateral migratory pulmonary infiltrates, with a predilection for the middle and lower lung zones, often with a minor-to-moderate pleural effusion. Lung function tests were usually normal; at the most a slight decrease in diffusing capacity was noted in some patients. There was no or only a slight response to antimicrobials; systemic corticosteroids were not given. Further evolution was benign with patients being asymptomatic between the episodes. Despite elaborate investigations, the cause of these 'pneumonias' remains frustratingly unknown. PMID- 11392576 TI - Serum-mediated relaxant response to toluene diisocyanate (TDI) in isolated guinea pig bronchi. AB - The present study was designed to evaluate whether pre-incubation with serum, obtained from both control and toluene diisocyanate (TDI)-immunized guinea-pigs, modified the contractile response to TDI in isolated guinea-pig bronchial rings. Guinea-pigs were anaesthetized and the main bronchi dissected in two rings. Bronchial rings were incubated with normal or immune serum (100 microl ml(-1) for 2 h) and dose-response curves to TDI (0.03-1000 microM) were studied isometrically. Before serum incubation, in eight bronchial rings, epithelium was removed by rubbing the luminal surface gently with a gauze. In control rings, TDI produced a concentration-dependent contraction, whereas in rings pre-incubated with either normal or TDI-immune serum, it produced a concentration-dependent relaxation. Relaxation was 101.4 (SEM 17.4)% and 94.9 (SEM 21)% of the relaxation induced by isoproterenol (1 mM) respectively with normal and TDI-immune serum. Similarly to the pre-incubation with serum, pre-incubation with albumin produced a concentration-dependent relaxation to TDI. Serum-induced relaxant response to TDI was not affected by capsaicin desensitization, it was only partially inhibited by an NK1-tachykinin antagonist, whereas it was blocked by indomethacin. In bronchial rings without epithelium, pre-incubated with serum, TDI caused contraction at highest doses, while it still induced relaxation at the lowest doses. This study shows that one or more components of the serum modify the contractile response to TDI in isolated guinea-pig bronchi. In bronchial rings without epithelium serum was able to inhibit the contration induced by low doses of TDI. PMID- 11392577 TI - Neutrophil-associated activation markers in healthy smokers relates to a fall in DL(CO) and to emphysematous changes on high resolution CT. AB - Smoking is a risk factor for developing chronic obstructive pulmonary disease (COPD), but there are no good indicators for early identification of subjects who will develop symptomatic COPD. The aim of this study was to investigate inflammatory mechanisms related to changes in lung function and emphysematous changes on high resolution computed tomography (HRCT) in 'healthy' smokers. Subjects were 60-year-old men from a population study. Bronchoscopy was performed in 30 smokers and 18 who had never smoked. Blood tests, lung function measurements and HRCT were carried out in 58 and 34 subjects, respectively. In comparison with never-smokers, smokers had higher levels of myeloperoxidase (MPO), human neutrophil lipocalin (HNL), eosinophil cationic protein (ECP) and lysozyme in blood, higher levels of MPO, interleukin-8 (IL-8) and HNL in bronchial lavage (BL), and of IL-8, HNL and interleukin-lbeta (IL-1beta) in bronchoalveolar lavage (BAL). Smokers also had lower levels of Clara cell protein 16 (CC-16) in blood. HNL in BL and BAL showed strong correlations to other inflammatory markers (MPO, IL-8, IL-1beta). The variations in MPO in BL were explained by variations in HNL (R2 =0.69), while these variations in BAL were explained by variations in HNL and IL-1beta (R2 = 0.76). DL(CO) was the lung function variable most closely related to MPO and IL-8 in BL and BAL and to IL 1beta in BAL. In a multiple regression analysis, MPO, IL-1beta, IL-8 and CC-16 in BL and MPO in BAL contributed to the explanation of variations in DL(CO) to 41% and 22%. respectively, independent of smoking habits. In smokers with emphysematous lesions on HRCT, HNL in BAL correlated to emphysema score (r(s) = 0.71). We conclude that 'healthy' smoking men with a near normal FEV1 show signs of inflammation in the lower airways that are related to a decrease in DL(CO) and to emphysematous lesions on HRCT. This inflammation seems to be the result of both monocyte/macrophage and neutrophil activation. PMID- 11392578 TI - Value and accuracy of cytology in addition to histology in the diagnosis of lung cancer at flexible bronchoscopy. AB - There is still disagreement as to the value and reliability of wash and brush cytology, in comparison with histology, for the diagnosis of malignancy at flexible bronchoscopy. The present study compares the yield and concordance of findings from the two modalities for visible tumours at flexible bronchoscopy. A single-centre study of 514 consecutive flexible bronchoscopy procedures, in which a lesion suspicious of cancer was seen and bronchial wash cytology, brush cytology and forceps biopsy samples were taken. All equivocal or suspicious results were taken as negative. An overall yield of 89.3% was achieved using a combination of all three tests. This was greater for endobronchial than submucosal (95% vs. 86%) tumours. Cytology alone diagnosed 17.7% of cases. Use of all three modalities allowed tumours to be differentiated between small and non small cell types in all but 5/459 positive cases (98.9%). There were only 3/313 cases in which there was a difference in cell type (small cell vs. non-small cell) between the two modalities. We conclude that wash and brush cytology are valuable tools, in addition to forceps biopsy, at flexible bronchoscopy. All three tests should be performed routinely in cases of suspected malignancy. PMID- 11392579 TI - Analysis of montelukast in mild persistent asthmatic patients with near-normal lung function. AB - Few studies have specifically evaluated controller therapy in patients with mild persistent asthma. We used a subgroup analysis to investigate the effects of montelukast, a potent cysteinyl leukotriene receptor antagonist, on adult patients on the milder end of the asthma severity spectrum. We have identified seven double-blind, randomized, placebo-controlled studies of adult patients with mild-to-moderate chronic asthma in which montelukast was investigated. Subsets of patients with baseline forced expiratory volume in 1 sec (FEV1) > 80% and > 75% predicted or further restricted by less than daily rescue beta-agonist use were included as four cohorts (A, B, C, D), and efficacy measures, including change in FEV1 rescue-free days, beta-agonist use, nocturnal awakenings and blood eosinophil counts were evaluated. Cohorts A to D comprised 21%, 8%, 11%, and 4%, respectively, of patients from these studies. Mean pretreatment FEV1 ranged from 81% to 84% predicted and daily beta-agonist use from 2.4 to 4.5 puffs day(-1) in the four cohorts. Pooled results demonstrated a treatment effect for montelukast over placebo in all cohorts, for all endpoints. There was a significant improvement in FEV1 in montelukast-treated patients (7-8% over baseline) compared with placebo (1-4% over baseline, between-group difference P < or = 0.02) for all cohorts. Similarly, the percentage of rescue-free days increased substantially more with montelukast (22-30%) than with placebo (8-13%). This subgroup analysis indicates that montelukast produced improvements in parameters of asthma control in patients with milder persistent asthma that should be confirmed in additional prospective trials. PMID- 11392581 TI - Optimal continuous positive airway pressure in patients with obstructive sleep apnoea: role of craniofacial structure. AB - Although nasal continuous positive airway pressure (CPAP) is effective in improving nocturnal obstructive apnoea, daytime sleepiness and well-being in patients with obstructive sleep apnoea syndrome (OSAS), not all patients tolerate this treatment. Since optimal CPAP titration is essential to maintain compliance, it is important to elucidate the factors that help to determine the optimal pressure. However, the determinants of the optimal CPAP level are controversial. The subjects comprised 27 Japanese male patients with OSAS who underwent standard polysomnography (PSG), pulmonary function tests, arterial blood gas analysis, cephalometry and CPAP titration. Twenty normal controls also underwent cephalometric analysis. The apnoea-hypopnoea index (AHI), mean oxygen saturation (mean SaO2) and the lowest SaO2 during sleep were found to be 54.7+/-22.6, 89.0+/ 5.6%, and 69.7+/-9.0%, respectively by PSG. The mean optimal CPAP was 9.6+/-1.8 cmH2O. The cephalometric angles (SNA, SNB and NSBa) were similar to those found in the control subjects. but MP-H, and PNS-P were significantly longer than those in the control subjects as shown by cephalometry. The optimal CPAP was correlated with the mean SaO2 (P<0.0001), neck circumference (P<0.05) and three cephalometric variables (NSBa: P<0.01, MP-H: P<0.05, PNS-P: P<0.05). Multiple, step-wise, regression analysis showed that the mean SaO2 and NSBa were independent variables that best predicted the optimal CPAP. These variables accounted for 57.5% of the total variance (R2=0.575, P<0.001). Optimal CPAP was closely correlated with oxygen desaturation during sleep. However, the craniofacial structure had additional effects such as an independent factor in determining the optimal CPAP level. PMID- 11392580 TI - Asthma and gastro-oesophageal reflux: can the response to anti-reflux therapy be predicted? AB - The aim of the study was to investigate which features predict favourable response to omeprazole therapy in asthmatics with gastro-oesophageal reflux (GER). The study population consisted of 52 outpatient asthmatics with GER who had completed an intervention where they were randomized to receive omeprazole 40 mg once a day or placebo for 8 weeks. After a 2-week washout period the patients were crossed over. Asthma symptoms were found to be relieved > or = 20% in 18 (35%) patients who were thus regarded as responders. A logistic regression analysis was performed in order to identify which features separate the responders from the non-responders. More responders were found among the patients whose body mass index (BMI) was higher (P = 0.02) or whose distal esophageal reflux was more severe [total time (%) pH < 4 (P = 0.01) or time (%) pH < 4 in upright position (P = 0.04)]. Adding other predictors to the total time (%) pH < 4, which was the most significant predictor for response in multi-variate analysis, did not further increase the prediction for favourable outcome. It is concluded that severe distal oesophageal reflux and obesity predict amelioration in asthma symptoms after 8-week omeprazole treatment in asthmatics with GER. Adding more than one predictor does not seem to further increase prediction for favourable asthma response. PMID- 11392582 TI - Has the perception of disability among COPD patients applying for pension changed during the last 20 years? AB - The aim is to examine the change in lung function, treatment and pulmonary symptoms in patients with chronic obstructive pulmonary disease (COPD) or chronic bronchitis (CB) applying for a pension during the period 1977-1996. In addition, we compared the perception of disability in males and females. From 1977 to 1996, 947 patients with COPD or CB were evaluated for obtaining economic support due to disability. In order to test the trend, the patients were divided into three periods: (1) 1977-1983, (2) 1984-1989 and (3) 1990-1996. Compared to females, males had substantial more pack-years of smoking (36 vs. 28, P<0.001), but their FEV1 was only slightly decreased (46.9% versus 49.6% predicted, P=0.047). Females reported significantly more often attacks of dyspnoea [OR: 1.5(1.00-2.2)] and any kind of dyspnoea during daytime [OR: 4.0(1.2-13.3)]. From period 1 to period 3, FEV1 increased significantly (45-53% predicted, P<0.001). Despite the increased FEV1, the use of inhaled corticosteroid had increased markedly (9-32% of the patients, P<0.001). The results did not change when patients with asthma were included. Our data suggest that both sexes, especially females, have become more aware of pulmonary symptoms and tend to react to them more actively by demanding evaluation and treatment. PMID- 11392583 TI - Compliance with inhaled glucocorticoids and concomitant use of long-acting beta2 agonists. AB - We investigated whether treatment with a long-acting beta2-agonist (LAbeta2) is associated with a decrease in patient compliance with regard to inhalation corticosteroids (ICS). Date on prescriptions collected by 15,760 patients suffering from airways disease were provided by 69 Dutch pharmacies. All prescriptions of ICS and LAbeta2 were analysed and divided in four groups by LAbeta2 use during 1997 and 1998. Date from 15,760 patients were available. In the 10,929 patients not treated with LAbeta2, compliance decreased slightly but not significantly. In 3281 patients receiving LAbeta2 compliance also decreased slightly but not significantly. In 404 patients, who used a LAbeta2 in 1997 and discontinued treatment in 1998, the compliance fell significantly (P<0.05). In 1147 patients who started to use a LAbeta2 in 1998, compliance with ICS significantly improved (P<0.05). These results suggest that the regular use of LAbeta2 improves compliance with ICS. Therefore, the concern that compliance with inhaled corticosteroid therapy will decrease under concomitant use of LAbeta2 appear to be unfounded. PMID- 11392584 TI - Effect of budesonide and nedocromil sodium on IL-6 and IL-8 release from human nasal mucosa and polyp epithelial cells. AB - We investigated the effect of budesonide and nedocromil sodium on the secretion of IL-6 and IL-8 by cultured epithelial cells from healthy nasal mucosa and nasal polyps. Human epithelial cell conditioned media was generated with fetal calf serum (FCS) in the presence or absence of budesonide and/or nedocromil sodium. Budesonide inhibited FCS-induced IL-6 and IL-8 release in a dose-dependent manner. The IC25 (25% inhibitory concentration) of budesonide on IL-6 release was higher in nasal polyp than in nasal mucosa epithelial cells (34 nM vs. 200 pM). The IC25 of budesonide on IL-8 release was higher in nasal mucosa than in nasal polyps (145 pM vs. 4 pM). Nedocromil sodium caused a dose-related inhibitory effect on IL-8 release from nasal mucosa (IC25, 207 nM), while it only had a significant effect in nasal polyps at 10(-5) M. Nedocromil sodium had no effect on IL-6 release. The inhibitory effect of budesonide was higher than that of nedocromil sodium on both nasal polyps and nasal mucosa. Budesonide and nedocromil sodium may exert their anti-inflammatory action in the respiratory mucosa by modulating the secretion of IL-6 and IL-8. The different effect of budesonide and nedocromil sodium on IL-6 and IL-8 release may be explained by differences in the mechanisms which regulate the upregulation of these cytokines in inflammatory responses. PMID- 11392585 TI - Diagostic value of respiratory impedance measurements in elderly subjects. AB - Obstructive lung disease (OLD) is highly prevalent in elderly subjects but markedly under-diagnosed. Indeed, only 40-50% of hospitalized elderly patients are able to adequately perform spirometric tests. This study aimed to evaluate, in an acute-care geriatric hospital, the diagnostic value of measuring airway impedance (Zrs) by the forced oscillation technique (FOT) for: (1) identifying OLD and (2) identifying responders vs. non-responders to bronchodilators. Sixty seven patients (aged 82+/-8 years) underwent consecutive measurement of Zrs and forced expiratory volumes before and after bronchodilators. Zrs was measured by FOT at frequencies of 4-30 Hz. Correlations, ROC curves and logistic regression models were established to determine the sensitivity (Se) and specificity (Sp) of Zrs in identifying OLD. Significant correlations were found between spirometric and Zrs measurements. The Zrs parameters yielding the best Se and Sp for detecting OLD were: Fn (resonant frequency; Se: 76%; Sp: 78%) and R0 (resistance extrapolated for a frequency of 0: Se: 76%; Sp: 74%). Using the logistic regression models, 76% of the patients were correctly classified as having OLD or not. Zrs was however not contributive in identifying responders to bronchodilators. Zrs measurements by FOT are contributive to the diagnosis of OLD in elderly hospitalized patients. PMID- 11392586 TI - Symptoms related to obstructive sleep apnoea are common in subjects with asthma, chronic bronchitis and rhinitis in a general population. AB - The purpose of this study was to examine the prevalence of self-reported snoring, apnoeas and daytime sleepiness in relation to chronic bronchitis, recurrent wheeze, physician-diagnosed asthma and rhinitis. This was a questionnaire study in a representative sample of a general population. The study was a part of the Obstructive Lung Disease in Northern Sweden Studies (OLIN). A total of 5424 subjects aged 20-69 years, born on the 15th day of each month, participated in the study. Eligible answers were obtained from 4648 subjects (85.7%). Having snoring as a problem was reported by 10.7%. Among subjects with chronic bronchitis it was reported by 25.9%, with recurrent wheeze by 21.3%, with physician-diagnosed asthma by 17.9%, and with rhinitis by 14.7%. Relatives' concerns of witnessed apnoea was reported by 6.8% of all subjects, while among subjects with chronic bronchitis it was reported by 18.1%, with recurrent wheeze by 17.1%, with physician-diagnosed asthma by 14.3%, and with rhinitis by 9.1%. After correction for age, gender and smoking habits, chronic bronchitis, rhinitis, asthma, and current smoking were significantly related, with snoring as a problem and with relatives' concern of witnessed apnoeas. Symptoms of daytime sleepiness were significantly related with concern of witnessed apnoeas, chronic bronchitis, snoring as a problem, recurrent wheeze and age 50-59 years. In conclusion, respiratory symptoms and conditions affecting mainly the lower respiratory tract, such as chronic bronchitis and asthma, were related with symptoms common in obstructive sleep apnoea. PMID- 11392587 TI - CCR2 and CCR5 gene polymorphisms in children with recurrent respiratory infections. PMID- 11392588 TI - Preterm delivery in patients admitted with preterm labor: a prediction study. AB - OBJECTIVE: To develop a model for prediction of preterm delivery in patients treated with parenteral tocolysis using combinations of maternal demographic and clinical factors. METHODS: We performed a retrospective cohort study using a perinatal database to identify women admitted with preterm labor and treated with parenteral tocolysis from 1980 to 1994. We developed an explanatory model using multiple logistic regression to determine the effect of four variables (prior preterm delivery, substance abuse, maternal complications and third-trimester care) on the likelihood of preterm delivery. For the prediction model, we initially included these four variables and then removed them in a stepwise fashion to determine the combination of the variables that offered the greatest model sensitivity and specificity. RESULTS: A total of 900 women were identified for the study and 247 (27%) had a preterm delivery. In the explanatory model, prior preterm delivery (OR 2.4; 95% CI 1.5-3.6), substance abuse (OR 2.2; 95% CI 1.2-5.1), initiation of care in the third trimester (OR 2.0; 95% CI 1.3-2.8) and medical complications of pregnancy (OR 1.8; 95% CI 1.2-2.6) increased the likelihood of preterm delivery. For the prediction tool, a three-variable model (prior preterm delivery, substance abuse and initiation of care in the third trimester) had high specificity (98%) and modest negative predictive value (73%). CONCLUSIONS: A simple three-variable model can correctly identify 98% of women with preterm labor treated with parenteral tocolysis who will not deliver preterm. Patients with no prior history of preterm delivery, no substance abuse and initiation of prenatal care before the third trimester have a 73% probability of not delivering preterm. PMID- 11392589 TI - Preterm premature rupture of membranes is not an independent risk factor for neonatal morbidity. AB - OBJECTIVE: To evaluate the risk factors for development of neonatal morbidity in cases of preterm premature rupture of membranes (PPROM). METHODS: The study population consisted of 2326 singleton preterm births occurring between 1994 and 1997 at Soroka University Medical Center. The neonatal morbidity included respiratory distress syndrome, intraventricular hemorrhage (grade III-IV), necrotizing enterocolitis, periventricular leukomalacia, bronchopulmonary dysplasia, neonatal pneumonia and sepsis. A cross-sectional study was designed to compare neonatal morbidity between two groups: the study group consisted of patients with PPROM (n = 376) and the comparison group of patients without PPROM (n = 1950). RESULTS: The prevalence of the neonatal morbidity associated with PPROM was 13.0% (49/376). There was no statistically significant difference in neonatal morbidity rates between the PPROM group and the group with intact membranes in any of the birth-weight groups (Mantel-Haenszel weighted odds ratio 1.20; 95% CI 0.80-1.20), or gestational-age groups (Mantel-Haenszel weighted odds ratio 1.03; 95% CI 0.79-1.55). There was no statistically significant difference in neonatal morbidity between patients with PPROM and those with intact membranes according to clinical chorioamnionitis. Congenital anomalies did not influence the neonatal morbidity when comparing patients with and without PPROM (44.4% vs. 32.8%, respectively; p = 0.23). CONCLUSIONS: PPROM was not an independent risk factor for neonatal morbidity in preterm births. Neonatal morbidity was affected mainly by prematurity itself, rather than by the occurrence of PPROM. PMID- 11392590 TI - Do maternal-fetal medicine practice characteristics influence high-risk referral decisions by general obstetrician-gynecologists? AB - OBJECTIVE: To determine whether the decision of the general obstetrician gynecologist to refer high-risk obstetric patients depends on the type of practice of the maternal-fetal medicine (MFM) specialist. METHODS: A questionnaire was mailed to 935 general obstetrician-gynecologists who were asked whether the MFM specialist's practice characteristics would influence their decision to refer their high-risk obstetric patients. Potential MFM practice components presented in the survey included: MFM, high-risk obstetrics, low-risk obstetrics or general obstetrics and gynecology. RESULTS: A total of 140 (15%) general obstetrician-gynecologists responded, 110 of whom were practicing obstetrics. Of the practicing responders, 77% stated that they were more likely to refer their high-risk obstetric patients if the MFM specialist practiced only MFM and high-risk obstetrics; 69% were less likely to refer their patients when the MFM specialist, in addition to MFM, practiced general obstetrics; and 75% were less likely to refer their patients when the MFM specialist also practiced general obstetrics and gynecology. The MFM practice setting (university vs. community hospital vs. private practice), as well as the geographic location and years of practice of the respondents, did not influence the general obstetrician gynecologists' decision to refer their high-risk obstetric patients. CONCLUSION: General obstetrician-gynecologists are more likely to refer high-risk obstetric patients if the MFM specialist practiced only MFM and high-risk obstetrics. PMID- 11392591 TI - Volumetric umbilical artery blood flow: comparison of the normal versus the single umbilical artery cord. AB - OBJECTIVE: This was a study of the volumetric blood flow in single umbilical artery (SUA) cords as compared to three-vessel cords. HYPOTHESIS: SUA flow will be twice that of an artery in a normal cord. METHODS: We studied 276 patients (24 SUA, 252 normal cord) at 18-40 weeks' gestation utilizing gray-scale and color Doppler. Flow, flow/kg, velocity, artery diameter, Doppler velocimetry indices, estimated fetal weight (EFW) and amniotic fluid index (AFI) were compared. All fetuses were anatomically and cytogenetically normal. RESULTS: Blood flow increased with advancing gestation and the SUA volume was twice that in the normal cord artery. Flow/kg decreased for both cords, with the SUA values twice those of normal cords. Arterial diameter and velocity increased, but to a greater degree in SUA. Velocimetry, although in the normal range, decreased progressively with the resistance indices always lower in the SUA cord. EFW and AFI were the same for both groups. CONCLUSION: Volumetric blood and its components were measured indirectly with ultrasound. The SUA cord artery carried twice the blood volume of an artery in a three-vessel cord. Other flow parameters changed appropriately to explain the increased flow. For the anatomically normal fetus with SUA there was no increase in intrauterine growth restriction. PMID- 11392592 TI - Characterization of growth-restricted fetuses with breakdown of the brain-sparing effect diagnosed by spectral Doppler. AB - OBJECTIVE: To characterize serial findings of the middle cerebral artery (MCA) and umbilical artery (UA) flow patterns, their relationship to each other, and neonatal outcomes in growth-restricted fetuses. METHODS: Serial pulsatility indices (PIs) from MCA and UA Doppler waveforms were measured in 41 growth restricted fetuses until Cesarean delivery. We found three patterns, as follows: phase 1 (n = 27), UA PI < MCA PI (no brain-sparing effect); phase 2 (n = 11), UA PI > MCA PI (brain-sparing effect); phase 3 (n = 3), both PIs elevated with the absence of end-diastolic flow or presence of reverse end-diastolic flow, which was designated as the 'breakdown of the brain-sparing effect'. Umbilical cord blood gas data at delivery were compared between each group. RESULTS: Age at delivery and body weights were significantly different for each phase. The mean body weights in all phases were significantly diminished from Japanese standard body weights, indicating growth restriction. The phase 3 pH and base excess were significantly different from those of the other two phases. CONCLUSIONS: Growth restricted fetuses which suffered from the state of breakdown of the brain sparing effect were delivered early with severe growth restriction and mild metabolic acidosis. The change from decreased to increased MCA PI along with increasing UA PI may predict a severely growth-restricted infant. PMID- 11392593 TI - University hospital-based prenatal care decreases the rate of preterm delivery and costs, when compared to managed care. AB - OBJECTIVE: Our purpose was to compare the rate of recurrent preterm delivery and the combined costs of mother/infant care for patients with a history of preterm delivery cared for in an inner city hospital house staff (HS) clinic versus an inner city managed care organization (MCO). METHODS: A retrospective cohort study was conducted. The groups consisted of 96 patients with a history of preterm delivery who were cared for by the HS clinic and 164 patients cared for in a neighborhood MCO. All patients with a history of previous preterm delivery who delivered at the Johns Hopkins Hospital between 1 January 1994 and 31 December 1996 were included in the study. The groups were examined regarding source of prenatal care for the subsequent pregnancy (HS vs. MCO), baseline demographics, intensity of prenatal care, maternal and neonatal outcomes and total cost of the provision of care. RESULTS: There were no differences in baseline demographics between the groups. There was a higher rate of recurrent prematurity for patients cared for by the MCO (HS, 24% vs. MCO, 36%, p = 0.04). Mean total mother/infant costs were also higher in the MCO group ($13565) when compared to the HS group ($9082), (p = 0.02). CONCLUSION: While some MCOs may provide cost savings for some low-risk populations, our study demonstrates that this health-care delivery system resulted in greater total expenditures for patients with a history of preterm delivery. PMID- 11392594 TI - Ursodeoxycholic acid for the treatment of intrahepatic cholestasis of pregnancy. AB - OBJECTIVE: To report our experience in managing intrahepatic cholestasis of pregnancy with ursodeoxycholic acid. METHODS: All cases of intrahepatic cholestasis of pregnancy that were diagnosed at Bridgeport Hospital from January 1997 to August 1999 were identified. Information was abstracted on demographics, medical and obstetric history, symptoms, laboratory data, therapy and pregnancy outcome. Statistical analysis was primarily descriptive; continuous variables were analyzed with t tests. RESULTS: A total of 20 cases of intrahepatic cholestasis of pregnancy were identified (0.32% of live births). All patients presented with pruritus. The mean gestational age at onset of symptoms was 31.1 weeks (range 13-38.4, median 32.4). Bile acids were measured in 18 cases and were elevated in all. The mean gestational age at delivery was 36.4 weeks (32.3-39.9). Eight patients were treated with ursodeoxycholic acid (600-1200 mg). All eight patients experienced subjective improvement in pruritus after initiation of treatment with ursodeoxycholic acid. Ursodeoxycholic acid was associated with a decrease in bile acids in most patients (p = 0.16) and with a significant decrease in serum transaminases (p = 0.03). CONCLUSIONS: Ursodeoxycholic acid is an effective therapy for relief of pruritus and improvement of the liver dysfunction that occurs with intrahepatic cholestasis of pregnancy. PMID- 11392595 TI - Effects of actinomycin D on fetal growth in rats. AB - OBJECTIVE: The goal of this study was to determine the effects of actinomycin D on fetal growth and prostaglandin dehydrogenase activity and to determine whether these effects could be obtained by administering extracts of fetal calf serum (SS 094). METHODS: Actinomycin D (7 microg/100 g body weight) was injected intraperitoneally on days 11 and 12 of pregnancy to induce growth restriction in rats. In another group, SS-094 was given (0.1 ml/100 g body weight) on days 12, 15, 17 and 19 of pregnancy, following the administration of actinomycin D. Placental prostaglandin dehydrogenase (PGDH) activity was measured on days 15 and 21 of pregnancy. Statistical analysis was performed by ANOVA and Dunnet's test. RESULTS: In the group injected with actinomycin D, fetal weight was significantly restricted (on day 15; p < 0.05; on day 21; p < 0.01), compared to the controls, and this restriction was successfully reversed by SS-094. PGDH activity in the placenta was significantly (p < 0.01) decreased in growth-restricted rats and remained low even after fetal weight recovered. CONCLUSIONS: Growth restriction in pregnant rats was successfully induced by actinomycin D, and SS-094 reversed this restriction. It does not seem that prostaglandin metabolism in the placenta is responsible for growth restriction. PMID- 11392596 TI - Rupture of an unscarred uterus with misoprostol induction: case report and review of the literature. AB - Uterine rupture is a serious and often tragic complication that is life threatening to both mother and child. It occurs at a frequency of around 1% in patients with a previously scarred uterus. Rupture of an unscarred uterus is an unexpected and devastating complication of pregnancy. With the increased use of misoprostol as a labor-inducing agent, cases of rupture of an unscarred uterus following its use have been published in the literature. We report a case of uterine rupture in a multigravid woman with an intrauterine fetal death at 29 weeks' gestation whose labor was induced with misoprostol. A review of all cases of uterine rupture with misoprostol induction is also included. Excessive doses of misoprostol should be used with extreme caution in multiparous women and in patients with a previously scarred uterus even in the context of intrauterine fetal death or termination of pregnancy. PMID- 11392597 TI - Guidelines and recommendations for safe use of Doppler ultrasound in perinatal applications. AB - Technological development has led to significant improvements in ultrasonographic capabilities in recent years, and this has been accompanied by increases in acoustic output. Meanwhile, there is a developing trend to use ultrasound at early stages of pregnancy when the developing embryo is known to be highly sensitive to damage by physical agents. The advent of pulsed spectral Doppler and color flow imaging has revolutionized perinatal applications. Doppler ultrasound has become widely accepted as a valuable diagnostic tool in obstetric medicine, where it has particular benefits for high-risk pregnancies. The benefits of Doppler screening are less well established. United States Food and Drug Administration (FDA) regulations now provide an option whereby equipment that provides a form of output display can be used to apply substantially higher acoustic output to the embryo or fetus than equipment approved for use under application-specific intensity limits. The Output Display Standard recently adopted by the FDA, in the USA, encourages self-regulation of acoustic exposure by the ultrasound user, on the basis of assumed knowledge of the implications of biophysical interactions. When modern sophisticated equipment is used at maximum operating settings for Doppler examinations, the acoustic outputs are sufficient to produce obvious biological effects, e.g. significant temperature increase in tissue or visible motion of particles due to radiation pressure streaming effects. The risk of inducing thermal effects is greater in the second and third trimesters, when fetal bone is intercepted by the ultrasound beam and significant temperature increase can occur in the fetal brain. Non-thermal bioeffects may be more significant in early gestation, when the relatively loosely tethered embryonic tissues are exposed to an ultrasound beam in a liquid path. The likelihood of producing cavitation-type non-thermal effects is enhanced by the presence in the sound-field of gas-encapsulated echo-contrast media. To ensure the continued safe use of ultrasound in obstetrics, it is important that international ultrasound organizations, such as the International Perinatal Doppler Society, issue advice to members to allow sensible assessment of risk: benefit and the practical implementation of the ALARA (as low as reasonably achievable) principle. PMID- 11392598 TI - Is high-dose misoprostol able to lower the incidence of cesarean section? A randomized controlled trial. AB - OBJECTIVE: To determine whether high-dose (100 microg) misoprostol was able to increase the rate of successful labor induction and lower the incidence of Cesarean section without adverse fetal effects. METHODS: A total of 360 women were randomized to receive either oxytocin (n = 192) by intravenous infusion, or misoprostol (n = 168) 100 microg intravaginally every 4 h. The Cesarean section rate was the primary end-point. Incidences of uterine and fetal heart rate abnormalities during labor and adverse neonatal outcomes were assessed as secondary end-points. RESULTS: Compared with those women receiving oxytocin, patients given misoprostol had a significantly shortened labor (10.7+/-6.0 vs. 15.4+/-10.4 h, p < 0.001). The Cesarean section rate did not differ between patients receiving misoprostol or oxytocin (36 (21.4%) vs. 38 (19.8%), p = 0.79) despite a sample size adequate to detect a 13 percentage point difference in this outcome. Patients receiving misoprostol had a higher incidence of the hyperstimulation syndrome (27 (16.1%) vs. 9 (4.7%), p < 0.001), and of fetal intolerance of labor as an indication for Cesarean delivery (23 (63.9%) vs. 15 (39.5%), p = 0.06), and had a greater number of umbilical artery cord blood pH findings of< 7.20 (20 (43.5%) vs. 6 (17.1%), p = 0.02). These worrisome trends on interim analysis resulted in our prematurely terminating the study. CONCLUSION: High-dose intravaginal misoprostol did not reduce the Cesarean section rate and was associated with a greater hazard of fetal intolerance of labor. PMID- 11392599 TI - Pregnancy outcomes in women with systemic lupus erythematosus. AB - OBJECTIVE: The purpose of this study was to examine pregnancy outcomes in women with systemic lupus erythematosus (SLE). STUDY DESIGN: Data from the California Health Information for Policy Project, which links records from birth certificates and hospital discharge records of mothers and newborns who delivered in all civilian hospitals in the state of California between 1 January 1993 and 31 December 1994, were retrospectively reviewed. Patients with a singleton gestation were stratified into the study group if they had a diagnosis of SLE, based on the International Classification of Disease, 9th Revision, or into the control group if they did not have SLE and delivered during the interval from 1 January 1994 to 31 December 1994. Specific maternal outcomes including pregnancy complications and fetal and neonatal outcomes were assessed and compared between the two groups. RESULTS: During the 2-year study period, 555 women had a diagnosis of SLE, and approximately 600000 women were included in the control group in the year 1994, giving a point prevalence of 0.05%. Specific adverse pregnancy outcomes, including hypertensive complications, renal disease, preterm delivery, non-elective Cesarean section, postpartum hemorrhage and delivery related deep vein thrombosis all occurred more frequently in the SLE group as compared to controls (p < 0.001). Additionally, neonatal and fetal outcomes were significantly worse in the SLE group, as documented by a higher prevalence of fetal growth restriction and neonatal death, as well as longer hospital stays (p < 0.0001). CONCLUSION: SLE was associated with a significant increase in maternal pregnancy complications and in fetal and neonatal morbidity and mortality as compared to the control population. However, our population-based study found significantly fewer adverse outcomes than were previously reported. This may represent a more accurate clinical picture of the impact of SLE on pregnancy outcomes. PMID- 11392600 TI - The effect of epidural analgesia on rates of episiotomy use and episiotomy extension in an inner-city hospital. AB - OBJECTIVE: To determine the relationship between epidural analgesia and episiotomy usage and episiotomy extension in parturients delivering vaginally. METHODS: A database of 20 888 women experiencing spontaneous vaginal delivery at Grady Memorial Hospital from 1990 to 1995 was examined to identify those receiving epidural analgesia. Patients who underwent epidural catheter placement and had adequate perineal anesthesia at delivery comprised the epidural group, and all others comprised the control group. Demographic characteristics and obstetric outcomes were compared. Univariate and multivariate analyses were used to test the association between epidural analgesia, rates of episiotomy and episiotomy extension. RESULTS: Of the 20888 women experiencing spontaneous vaginal deliveries 6785 (32.5%) received epidural analgesia. Women receiving epidural analgesia were more likely than those not receiving epidural analgesia to be African-American and nulliparous, and to have an occiput posterior presentation. Women receiving epidural analgesia were also more likely to receive an episiotomy (27.8% vs. 13.1%, odds ratio (OR) 2.56, 95% confidence interval (CI) 2.38-2.75) and were less likely to experience a second-degree perineal laceration (11.6% vs. 14.4%, OR 0.75, 95% CI 0.69-0.82) or a third- or fourth degree extension (8.9% vs. 12.4%, OR 0.81, 95% CI 0.68-0.97). When the results were adjusted for nulliparity, posterior presentation, macrosomia, shoulder dystocia and prolonged second stage, epidural analgesia remained independently associated with receipt of episiotomy (OR 1.97, 95% CI 1.88-2.06) and reduced episiotomy extension (OR 0.74, 95% CI 0.54-0.94). CONCLUSION: Epidural analgesia increases the rates of episiotomy use, and decreases the rate of episiotomy extension, independently of clinical factors associated with episiotomy. PMID- 11392601 TI - Individual prediction of treatment outcome in patients with temporomandibular disorders. AB - Out of 3159 patients referred to a specialist clinic of stomatognathic physiology, 1297 patients started treatment because of temporomandibular disorders. All those patients were divided into subgroups of patients with mainly muscular or TMJ symptoms, respectively. A prediction of the treatment outcome as good or dubious was also made. This prediction was based on the patient's history and the clinical findings. A total of 989 patients (76%) completed the treatment. They all graded the degree of improvement of their initial symptoms using a Numeric Rating Scale. An improvement of initial complaints of 50% or more was judged to be a relevant change. In 85% of the patients with mainly muscular symptoms, the treatment was predicted to have a good outcome. The corresponding figures for those patients with mainly TMJ symptoms were 93%. Not less than 88% of all the patients reported an improvement of 50% or more of their initial complaints. The possibility to predict the treatment outcome as good in patients with mainly muscular symptoms was very good: 90% fulfilled the criteria of a positive treatment result. For those with a dubious prognosis, 56% had a positive outcome of treatment. The possibility to predict treatment outcome as good in those with mainly TMJ symptoms was also very good: 94% responded positively on the treatment. Almost the same figure, 88%, of those with a poor prognosis had a positive treatment outcome. PMID- 11392602 TI - Prevalence of contact dermatitis among dental personnel in a Swedish rural county. AB - The use of acrylate-based polymers in dental care in Sweden has increased dramatically in recent years. Acrylate monomers are sensitizers and can cause contact dermatitis. Allergy to latex, e.g. in rubber gloves, is also an increasing problem in health care and especially in dental care. The aim was to estimate the prevalences of work related allergies in dental personnel in a Swedish county with almost 700 dentists, dental nurses and dental hygienists. A questionnaire was distributed to all dental care units in the county and 98% of the population completed it. Specific allergy was verified with standard patch and prick testing. Work related allergy symptoms were reported by 189 subjects, 28%. The prevalences of acrylate eczema and latex allergy were 3,0% and 3,7% respectively. The prevalence of all work related skin allergy was estimated to 8%. The estimated prevalence of acrylate allergy was close to those of a few other published reports. However, the prevalence of hypersensitivity to latex rubber was lower than those reported from other studies. It is concluded that almost one dental care worker out of ten has a risk of contact dermatitis, unless measures to reduce hazardous exposure have been taken. PMID- 11392603 TI - Resin-bonded bridges by dental undergraduates: three-year follow-up. AB - This study was conducted to evaluate performance and longevity of resin-bonded bridges in patients treated by undergraduate dental students. A total of 62 bridges had been constructed for 53 patients by the students at the School of Dentistry, Karolinska Institute, between 1989 and 1997. After a mean observation period of 35.3 months retention had been lost in 13.6% of 59 bridges examined (4.6% per year). The debonding rate was greater in the mandible (8.3 %/year) than in the maxilla (2.7 %/year), and greatest in the mandibular anterior region (13.4%). Thirty-nine patients with 46 bridges (27 maxillary and 19 mandibular bridges), underwent more detailed clinical examination. No significant differences in bleeding and pocket depth were found between abutment teeth and controls. Bleeding was more frequent at the approximal than the lingual surfaces of the abutment teeth. A questionnaire revealed high patient satisfaction, the resin-bonded bridges meeting their expectations. The study suggests that resin bonded bridges are an acceptable alternative to conventional bridges. Successful outcome is dependent on stringent case selection, an abutment preparation designed to establish retention form, and meticulous bonding procedures. PMID- 11392604 TI - Benzodiazepines in child dental care: a survey of its use among general practitioners and paediatric dentists in Sweden. AB - Dental treatment in children too young or too apprehensive to co-operate is often performed under sedation. The aim of this study was to survey the use of rectal and oral liquid and tablet benzodiazepine sedation in Swedish child dentistry, and estimation of treatment success. A questionnaire was sent to 500 randomly selected dentists (GPs) working in the Public Dental Health Service and all (77) specialists (PDs) working at paediatric dentistry clinics. Benzodiazepine sedation was used by 73% of the GPs and 97% of the PDs. Seven per cent of the GPs and 87% of the PDs had sedation sessions at least once a month. Of the GPs, 60% administered the sedation rectally, 7% orally in liquid form, and 39% orally in tablet form. For PDs, the corresponding figures were 97%, 78%, and 68%. Sixteen per cent of the GPs and 84% of the PDs used midazolam for rectal sedation. PDs rated rectal sedation better than the GPs (p < 0.001). GPs rated their experiences of rectal sedation as better the more frequent the use (p = 0.03), as did PDs concerning oral liquid sedation (p = 0.03). Thus, it seems that a more regular use of sedation is advantageous in achieving better treatment outcome. PMID- 11392605 TI - A review of oral devices in the treatment of habitual snoring and obstructive sleep apnoea. AB - With recent interest in sleep apnoea, oral devices have been proposed and used increasingly to treat patients suffering from snoring and obstructive sleep apnoea (OSA). Numerous case reports have been published and studies have been carried out to determine the efficiency and mechanism of action as well as evaluate side effects, complications and costs of different oral devices in the treatment of habitual snoring and OSA. A summary of the scientific basis and current opinions regarding oral devices in the treatment of habitual snoring and obstructive sleep apnoea is presented in this article. The selection of papers was based on a computerised search of published clinical and associated studies identified by MEDLINE from 1980 to November 2000. Although there is not yet enough scientific evidence for the clinician to determine which appliance is most likely to improve symptoms for a given patient, it appears from the literature that dental devices may have a place in the treatment of habitual snoring and mild and moderate cases of OSA. However, most of the studies are case series, raising questions on validity, and few studies use appropriate control groups. The general opinion is that these patients should not be treated without a sleep study as the OSA must be diagnosed before beginning treatment with oral devices to identify patients at risk and to provide a baseline to establish the effectiveness of the treatment. The main advantages of oral devices is the relative simplicity of the treatment, its reversibility and cost-effectiveness, and the fact that they can be used as an alternate treatment in patients who are unable to tolerate nasal continuous positive airway pressure (nCPAP) or who are poor surgical risks. Side effects occur in a significant proportion of patients using the MAD. In most cases these are minor and their importance must be balanced against the efficacy of the MAD in treating snoring and OSA. n. PMID- 11392606 TI - Selective matrix metalloproteinase (MMP) inhibition in rheumatoid arthritis- targetting gelatinase A activation. AB - The matrix metalloproteinases (MMP) are a large group of enzymes responsible for matrix degradation. They contribute to joint destruction in rheumatoid arthritis (RA) by directly degrading the cartilage and bone and indirectly promoting angiogenesis (formation of new blood vessels). Inhibition of MMPs is a primary therapeutic target in RA. However, the results of limited clinical trials performed to date are disappointing. Improvements in therapeutic benefit may be achieved by targetting specific MMPs. A subclass of the MMPs, the gelatinases, contribute directly to joint destruction as well as being vital during angiogenesis. Gelatinase A is released as a latent enzyme and must be activated to degrade the matrix. It has a unique mechanism of activation on the cell surface involving membrane-type MMP (MT-MMP). Recently, the serine protease, activated protein C (APC), has been shown to directly activate gelatinase A, without requiring MT-MMP Inhibition of APC represents a selective approach to prevent gelatinase A activation and may prove to be of therapeutic benefit in RA. PMID- 11392607 TI - Granulocyte-colony stimulating factor in the prevention of postoperative infectious complications and sub-optimal recovery from operation in patients with colorectal cancer and increased preoperative risk (ASA 3 and 4). Protocol for a controlled clinical trial developed by consensus of an international study group. Part two: design of the study. AB - GENERAL DESIGN: Presentation of a new type of a study protocol for evaluation of the effectiveness of an immune modifier (rhG-CSF, filgrastim): prevention of postoperative infectious complications and of sub-optimal recovery from operation in patients with colorectal cancer and increased preoperative risk (ASA 3 and 4). This part describes the design of the randomised, placebo controlled, double blinded, single-centre study performed at an university hospital (n = 40 patients for each group). OBJECTIVE: The trial design includes the following elements for a prototype protocol: * The study population is restricted to patients with colorectal cancer, including a left sided resection and an increased perioperative risk (ASA 3 and 4). * Patients are allocated by random to the control or treatment group. * The double blinding strategy of the trial is assessed by psychometric indices. * An endpoint construct with quality of life (EORTC QLQ-C30) and a recovery index (modified Mc Peek index) are used as primary endpoints. Qualitative analysis of clinical relevance of the endpoints is performed by both patients and doctors. * Statistical analysis uses an area under the curve (AUC) model for improvement of quality of life on leaving hospital and two and six months after operation. A confirmatory statistical model with quality of life as the first primary endpoint in the hierarchic test procedure is used. Expectations of patients and surgeons and the negative affect are analysed by social psychological scales. CONCLUSION: This study design differs from other trials on preoperative prophylaxis and postoperative recovery, and has been developed to try a new concept and avoid previous failures. PMID- 11392608 TI - Mouse macrophages release a neutrophil chemotactic mediator following stimulation by staphylococcal enterotoxin type A. AB - OBJECTIVE AND DESIGN: To examine the role of macrophages in the neutrophil migration induced by staphylococcal enterotoxin type A (SEA) in mice. MATERIALS AND METHODS: Peritoneal macrophages were harvested from male Swiss mice pre treated with thioglycollate. After adhering to plastic tissue culture dishes, the cells were washed and incubated with RPMI or SEA (0.62-2.5 microg/ml) and washed again prior to further incubation with RPMI alone. The medium was then collected, sterilized and assayed for promigratory activity in the mouse peritoneal cavity. RESULTS: Mouse macrophage monolayers stimulated with SEA secreted a thermolabile neutrophil chemotactic component (MNCC-SEA) with a molecular mass >100 kDa (by ultrafiltration). This release was dose- and time-dependent and was inhibited by dexamethasone but not by indomethacin or BW755C. Dexamethasone, indomethacin, BWA4C, BW755C, BN52021, cimetidine and SR48968 had no effect on the neutrophil migration induced by MNCC-SEA while capsaicin and SR 140333 reduced this phenomenon. CONCLUSIONS: Macrophages play a key role in the neutrophil recruitment induced by SEA probably by releasing an MNCC-SEA that presumably induces neutrophil migration via a mechanism mediated by substance P. PMID- 11392610 TI - Calcium-dependent interleukin-8 gene expression in T84 human colonic epithelial cells. AB - OBJECTIVE AND DESIGN: IL-8 is a chemokine that activates and recruits neutrophils and plays a major role in intestinal inflammation. Signal transduction pathways mediated by protein kinases are central in regulating IL-8 gene expression, however, little is known about the role of Ca2+ in this event. In this study, we characterize the effect of intracellular Ca2+ on interleukin-8 gene expression in T84 human colonic epithelial cells. MATERIALS AND METHODS: Cells were stimulated with Ca2+ ionophore, A23187 or thapsigargin, a Ca2+-ATPase inhibitor. Semi quantitative RT-PCR was used to examine IL-8 mRNA and ELISA for protein quantification. Reporter gene techniques were used to determine transcription rate. RESULTS: A23187 and thapsigargin caused a dose- and time-dependent accumulation of IL-8 mRNA and protein production which was dependent on the release of Ca2+ from intracellular stores. FK506, a specific inhibitor of calcineurin, inhibited A23187- and thapsigargin-induced IL-8 mRNA expression in a dose dependent manner. Reporter gene studies and actinomycin D chase experiments showed that A23187 and thapsigargin enhanced IL-8 gene transcription and stabilized IL-8 mRNA transcripts, respectively. CONCLUSION: Intracellular Ca2+ plays an important role in regulating IL-8 transcriptionally and posttranscriptionally through calcium/calmodulin-dependent calcineurin. PMID- 11392609 TI - The water-soluble components of the essential oil of Melaleuca alternifolia (tea tree oil) suppress the production of superoxide by human monocytes, but not neutrophils, activated in vitro. AB - OBJECTIVE: To evaluate the regulatory properties of the essential oil of Melaleuca alternifolia (tea tree oil) on the production of oxygen derived reactive species by human peripheral blood leukocytes activated in vitro. MATERIALS AND METHODS: The ability of tea tree oil to reduce superoxide production by neutrophils and monocytes stimulated with N-formyl-methionyl-leucyl phenylalanine (fMLP), lipopolysaccharide (LPS) or phorbol 12-myristate 13-acetate (PMA) was examined. RESULTS: The water-soluble fraction of tea tree oil had no significant effect on agonist-stimulated superoxide production by neutrophils, but significantly and dose-dependently suppressed agonist-stimulated superoxide production by monocytes. This suppression was not due to cell death. Chemical analysis identified the water-soluble components to be terpinen-4-ol, alpha terpineol and 1,8-cineole. When examined individually, terpinen-4-ol significantly suppressed fMLP- and LPS- but not PMA-stimulated superoxide production; alpha-terpineol significantly suppressed fMLP-, LPS- and PMA stimulated superoxide production; 1,8-cineole was without effect. CONCLUSION: Tea tree oil components suppress the production of superoxide by monocytes, but not neutrophils, suggesting the potential for selective regulation of cell types by these components during inflammation. PMID- 11392611 TI - Serum IgE specific to indoor moulds, measured by basophil histamine release, is associated with building-related symptoms in damp buildings. AB - OBJECTIVE: To study the relationship between basophil histamine release (HRT) to indoor moulds, indicating specific IgE, and building-related symptoms (BRS), asthma, and hay fever in individuals working in damp and mouldy buildings. METHODS: A cross-sectional study was performed among 86 school staff members, who on average had worked 143 months (range: 3-396) in moist buildings with mould growth in the constructions. A questionnaire concerning mucous membrane symptoms, facial skin symptoms, central nervous system symptoms, hay fever, and asthma was fulfilled by the participants, and blood samples were taken. Eight mould species growing on building constructions were identified and cultivated to obtain allergenic materials for testing. The presence in serum of IgE specific to moulds was verified by histamine release test (HRT) based on passive sensitization of basophil leukocytes. The validity of the method was confirmed by parallel testing of patients allergic to grass- and birch pollen and by the shift from positive to negative response after removal of serum IgE and by using sham sensitization. RESULTS: The prevalence of most BRS was between 32% and 62%. Positive HRT, showing serum IgE specific to one or more of the moulds, was observed in 37% of the individuals. The highest frequency of positive HRT was found to Penicillium chrysogenum and then to Aspergillus species, Cladosporium sphaerospermum and Stachybotrys chartarum. A significant association was found between most BRS and positive HRT, whereas no association was observed between positive HRT to moulds and self reported hay fever or asthma. CONCLUSION: Positive HRT to indoor moulds, showing the presence in serum of IgE specific to the fungi, was found to be related to BRS in individuals working in damp and mouldy buildings. Whether the association is of causal character is a question for further studies. The test may be useful in the evaluation and study of possible mould induced BRS. PMID- 11392612 TI - M5 muscarinic receptors are needed for slow activation of dopamine neurons and for rewarding brain stimulation. AB - Mesopontine cholinergic neurons (Ch5 and Ch6 cell groups) activate the cerebral cortex via thalamic projections, and activate locomotion and reward via dopamine neurons in the substantia nigra and ventral tegmental area (VTA). Nicotinic receptors in VTA activate dopamine neurons quickly, and are needed for the stimulant and rewarding effects of nicotine in rats. Muscarinic receptors in VTA activate dopamine neurons slowly, and are needed for the rewarding effects of hypothalamic stimulation, but do not increase locomotion. Antisense oligonucleotides targetting M5 mRNA, when infused into the VTA, inhibited M5 receptor binding and rewarding hypothalamic stimulation. Mutant mice with truncated M5 muscarinic receptor genes drank more water than wild-type controls. Spontaneous locomotion and locomotor responses to amphetamine and scopolamine were unchanged. Electrical stimulation near Ch6 induced dopamine release in the nucleus accumbens in two phases, an early phase (0-2 min after stimulation) dependent on nicotinic and gluatamatergic receptors in VTA, and a late phase (8 50 min after stimulation) dependent on muscarinic receptors in VTA. The late phase was lost in M5 mutant mice, while the early phase was unchanged. M5 muscarinic receptors bind slowly to muscarinic ligands, and appear to mediate slow secretions. PMID- 11392613 TI - Generation and pharmacological analysis of M2 and M4 muscarinic receptor knockout mice. AB - Muscarinic acetylcholine receptors (M1-M5) play important roles in the modulation of many key functions of the central and peripheral nervous system. To explore the physiological roles of the two Gi-coupled muscarinic receptors, we disrupted the M2 and M4 receptor genes in mice by using a gene targeting strategy. Pharmacological and behavioral analysis of the resulting mutant mice showed that the M2 receptor subtype is critically involved in mediating three of the most striking central muscarinic effects, tremor, hypothermia, and analgesia. These studies also indicated that M4 receptors are not critically involved in these central muscarinic responses. However, M4 receptor-deficient mice showed an increase in basal locomotor activity and greatly enhanced locomotor responses following drug-induced activation of D1 dopamine receptors. This observation is consistent with the concept that M4 receptors exert inhibitory control over D1 receptor-mediated locomotor stimulation, probably at the level of striatal projection neurons where the two receptors are known to be coexpressed. These findings emphasize the usefulness of gene targeting approaches to shed light on the physiological and pathophysiological roles of the individual muscarinic receptor subtypes. PMID- 11392614 TI - Role of the M1 receptor in regulating circadian rhythms. AB - Cholinergic stimuli are potent regulators of the circadian clock in the hypothalamic suprachiasmatic nucleus (SCN). Using a brain slice model, we have found that the SCN clock is subject to muscarinic regulation, a sensitivity expressed only during the night of the clock's 24-h cycle. Pharmacological and signal transduction characteristics are compatible with a response mediated by an M1-like receptor. Molecular manipulation of muscarinic receptors will provide important insights as to the receptor subtype(s) regulating circadian rhythms. PMID- 11392615 TI - Investigations into the physiological role of muscarinic M2 and M4 muscarinic and M4 receptor subtypes using receptor knockout mice. AB - Determination of muscarinic agonist-induced parasympathomimetic effects in wild type and M2 and M4 muscarinic receptor knockout mice revealed that M2 receptors mediated tremor and hypothermia, but not salivation. The M4 receptors seem to play a modest role in salivation, but did not alter hypothermia and tremor. In the M2 knockout mice, agonist-induced bradycardia in isolated spontaneously beating atria was completely absent compared to their wild type litter mates, whereas agonist-induced bradycardia was similar in the M4 knockout and wild type mice. The potency of carbachol to stimulate contraction of isolated stomach fundus, urinary bladder and trachea was reduced by a factor of about 2 in the M2 knockout mice, but was unaltered in the M4 knockout mice. The binding of the muscarinic agonist, [3H]-oxotremorine-M, was reduced in cortical tissue from the M2 knockout mice and to a lesser extent from the M4 knockout mice, and was reduced over 90% in the brain stem of M2 knockout mice. The data demonstrate the usefulness of knockout mice in determining the physiological function of peripheral and central muscarinic receptors. PMID- 11392616 TI - Identification of subtypes of muscarinic receptors that regulate Ca2+ and K+ channel activity in sympathetic neurons. AB - Many different G protein-coupled receptors modulate the activity of Ca2+ and K+ channels in a variety of neuronal types. There are five known subtypes (M1-M5) of muscarinic acetylcholine receptors. Knockout mice lacking the M1, M2, or M4 subtypes are studied to determine which receptors mediate modulation of voltage gated Ca2+ channels in mouse sympathetic neurons. In these cells, muscarinic agonists modulate N- and L-type Ca2+ channels and the M-type K+ channel through two distinct, G-protein mediated pathways. The fast and voltage-dependent pathway is lacking in the M2 receptor knockout mice. The slow and voltage-independent pathway is absent in the M1 receptor knockout mice. Neither pathway is affected in the M4 receptor knockout mice. Muscarinic modulation of the M current is absent in the M1 receptor knockout mice, and can be reconstituted in a heterologous expression system using cloned channels and M1 receptors. Our results using knockout mice are compared with pharmacological data in the rat. PMID- 11392617 TI - Alteration of cardiovascular and neuronal function in M1 knockout mice. AB - We used gene targeting to generate mice lacking the M1 muscarinic acetylcholine receptor. These mice exhibit a decreased susceptibility to pilocarpine-induced seizures, loss of regulation of M-current potassium channel activity and of a specific calcium channel pathway in sympathetic neurons, a loss of the positive chronotropic and inotropic responses to the novel muscarinic agonist McN-A-343, and impaired learning in a hippocampal-dependent test of spatial memory. PMID- 11392618 TI - The conformational switch in muscarinic acetylcholine receptors. AB - The recently-determined structure of rhodopsin has provided a suitable basis for modeling the three-dimensional structure of the M1 muscarinic acetylcholine receptor. Using this as a framework for interpreting mutagenesis studies, we have been able to suggest most of the contacts which the receptor makes with acetylcholine and many of the intramolecular contacts which are important for the ground-state structure of the receptor. It is possible to outline a mechanism of G-protein interaction. PMID- 11392619 TI - Dual effects of muscarinic M2 receptors on the synthesis of cyclic AMP in CHO cells: background and model. AB - It has been observed in several laboratories that muscarinic agonists have dual effects on the synthesis of cyclic AMP in cell lines expressing muscarinic M2 or M4 receptors, producing strong inhibition at low agonist concentrations and lesser inhibition or stimulation at high agonist concentrations. Data obtained on CHO cells (known to express adenylyl cyclases VI and VII) are best interpreted on the assumption that the upward phase of the concentration-response curves reflects simultaneous inhibition of adenylyl cyclase VI via the Gi proteins, with which the M2 and M4 receptors communicate with high affinity, and stimulation of adenyly cyclases VI and VII via the Gs proteins, with which the M2 and M4 receptors communicate with low affinity. A simplified model is described which permits one to predict how the shapes of the concentration-response curves will be affected by changes in the concentration of receptors, the affinities of activated receptors for Gi or Gs proteins, and other parameters. PMID- 11392620 TI - Constitutively active muscarinic receptors. AB - Mutations that increase constitutive activity and alter ligand binding have been used to investigate the structure and mechanism of activation of muscarinic receptors. These data are reviewed with reference to the recently published three dimensional structure of rhodopsin. Residues in TM3 and TM6 where amino acid substitutions increased constitutive activity align with residues within the core of the receptor. A nucleus of these residues is located immediately below the predicted binding site of acetylcholine. The i2 loop where mutations also increase constitutive activity was found to loop away from the i3 loop, which has been found to modulate G-protein coupling specificity. PMID- 11392621 TI - Multiple allosteric sites on muscarinic receptors. AB - Proteins and small molecules are capable of regulating the agonist binding and function of G-protein coupled receptors by multiple allosteric mechanisms. In the case of muscarinic receptors, there is the well-characterised allosteric site that binds, for example, gallamine and brucine. The protein kinase inhibitor, KT5720, has now been shown to bind to a second allosteric site and to regulate agonist and antagonist binding. The binding of brucine and gallamine does not affect KT5720 binding nor its effects on the dissociation of [3H]-N methylscopolamine from M1 receptors. Therefore it is possible to have a muscarinic receptor with three small ligands bound simultaneously. A model of the M1 receptor, based on the recently determined structure of rhodopsin, has the residues that have been shown to be important for gallamine binding clustered within and to one side of a cleft in the extracellular face of the receptor. This cleft may represent the access route of acetylcholine to its binding site. PMID- 11392622 TI - Novel signalling events mediated by muscarinic receptor subtypes. AB - The M2 muscarinic acetylcholine receptor (mAChR) activates Gi protein coupled pathways, such as stimulation of G-protein activated inwardly rectifying K channels (GIRKs). Here we report a novel heterologous desensitization of these GIRK currents, which appeared to be specifically induced by M2/M4 mAChR stimulation, but not via adenosine (Ado) and alpha2-adrenergic receptors (AR). This heterologous desensitization reflected an inhibition of the GIRK signalling pathway downstream of G-protein activation. It was mediated in a membrane delimited fashion via a PTX insensitive GTP dependent pathway and could be competed with exogenous Gbetagamma. The activation of M3 mAChR/Gq coupled receptors potently inhibited GIRK currents similar as M2 mAChR. By monitoring simultaneously the response of A1 adenosine receptor (AdoR) activation on N-type Ca2+ channels and GIRK channels, the stimulation of M3 mAChR was found to cause an inhibition of the Ado response in both effector systems, suggesting that the inhibition occurred at the level of the G-protein common to both effectors. These results indicated that Gq proteins inhibit pathways that are commonly regulated by Gbetagamma proteins. PMID- 11392623 TI - Sphingosine kinase-mediated calcium signaling by muscarinic acetylcholine receptors. AB - Based on the finding that G protein-coupled receptors (GPCRs) can induce Ca2+ mobilization, apparently independent of the phospholipase C (PLC)/inositol-1,4,5 trisphosphate (IP3) pathway, we investigated whether sphingosine kinase, which generates sphingosine-1-phosphate (SPP), is involved in calcium signaling by mAChR and other GPCRs. Inhibition of sphingosine kinase by DL-threo dihydrosphingosine and N,/N-dimethylsphingosine markedly inhibited [Ca2+]i increases elicited by M2 and M3 mAChRs in HEK-293 cells without affecting PLC activation. Activation of M2 and M3 mAChR rapidly and transiently stimulated production of SPP. Furthermore, microinjection of SPP into HEK-293 cells induced rapid and transient Ca2+ mobilization. Pretreatment of HEK-293 cells with the calcium chelator BAPTA/AM fully blocked mAChR-induced SPP production. On the other hand, incubation of HEK-293 cells with calcium ionophores activated SPP production. Similar findings were obtained for formyl peptide and P2Y2 purinergic receptors in HL-60 cells. On the basis of these studies we propose, that following initial IP3 production by receptor-mediated PLC activation, a local discrete increase in [Ca2+]i induces sphingosine kinase stimulation, which ultimately leads to full calcium mobilization. Thus, sphingosine kinase activation most likely represents an amplification system for calcium signaling by mAChRs and other GPCRs. PMID- 11392624 TI - Snake toxins that bind specifically to individual subtypes of muscarinic receptors. AB - This paper discusses the properties of the three most specific ligands found for the extracellular faces of M1, M2 and M4 muscarinic receptors (m1-toxin1, m2 toxin and m4-toxin, respectively). The primary goal of this paper is to show the known and potential usefulness of these toxins and their biotinylated, radioactive, fluorescent and mutated derivatives. PMID- 11392625 TI - Clinical experiences with tolterodine. AB - Tolterodine is the first muscarinic receptor antagonist that has been specifically developed for the treatment of overactive bladder. The objectives in the discovery program were to design a potent muscarinic receptor antagonist that is equipotent to oxybutynin in the bladder, but less potent in salivary glands, with the aim of improving tolerability (less dry mouth) in patients with overactive bladder. Tolterodine is non-selective with respect to the muscarinic M1-M5 receptor subtypes, but has a greater effect on the bladder than on salivary glands in vivo, in both animals and humans. Clinical results show that the efficacy and safety of tolterodine in overactive bladder is equal to that of oxybutynin, but that tolterodine is significantly better tolerated by the patients. PMID- 11392626 TI - Antimuscarinic treatment for lung diseases from research to clinical practice. AB - Inhaled antimuscarinic drugs are the treatment of choice, recommended by guidelines, in chronic obstructive pulmonary disease (COPD). In long-term clinical studies ipratropium shows important effects beyond relaxation of airway smooth muscle, e.g. reduction of exacerbations of COPD. In phase III clinical trials the new generation antimuscarinic tiotropium, inhaled once daily, has provided more than 24 hours of stable bronchodilation, that was sustained over the one year treatment period. In addition, tiotropium in comparison to placebo and even ipratropium, has been shown to provide improvement in dyspnea, reduction of exacerbations of COPD, reduced hospital admissions for exacerbations, reduced duration of hospitalisations as well as improved health-related quality of life. Chronic effects, such as reduction of hospitalisations, are conventionally attributed to an anti-inflammatory action and not to symptomatic bronchodilation. The 24 hour stabilisation of airway patency, avoiding fluctuations of the diameter with occasional closure and consequent need for reopening, may explain the extended therapeutic profile of tiotropium. Inhibition by antimuscarinics of pro-inflammatory cholinergic effects may also occur, e.g. inhibition of 5-HETE release from epithelial cells and inhibition of release of neutrophil and eosinophil chemotactic activity from alveolar macrophages. Antimuscarinics have shown increasing value as a therapeutic approach in COPD. The elucidation of their anti-inflammatory potential constitutes an interesting target for future studies. PMID- 11392627 TI - Anticholinergic therapy for airway diseases. AB - Anticholinergics are commonly used in the treatment of airway diseases. While their effectiveness in chronic asthma offers no advantage over beta-agonists, evidence continues to accumulate suggesting substantial additional benefit in acute asthma attacks. This increased response to anticholinergics suggest that cholinergic bronchoconstriction is increased in acute asthma. Multiple mechanisms related to changes in expression and function of inhibitory M2 muscarinic receptors on the airway parasympathetic nerves may be involved, and are discussed. The use of anticholinergics in chronic obstructive pulmonary disease and in rhinitis are also considered. PMID- 11392628 TI - Muscarinic receptors and gastrointestinal tract smooth muscle function. AB - Over the last decade, several lines of evidence have shown that both muscarinic M2 and M3 receptors are postjunctionally expressed in many smooth muscles, including the gastrointestinal tract. Although in vitro data suggests that both receptors are functional in that they inhibit adenylate cyclase activity and activate non-selective cation channels, few studies support a role in vivo. Thus, data from procedures that ablate the signaling pathway of the muscarinic M2 receptor, including receptor antagonism, pertussis toxin pretreatment reveal little effect on gastrointestinal smooth muscle responsiveness to muscarinic agonists. Recently, information from knockout mice, lacking either M2 or M3 receptor, indicate reveal a role for both subtypes. However, the contribution of the M2 receptor appears greater in the ileum than in the urinary bladder. Therapeutically, non-selective, as well as selective M3 receptor antagonists are being clinically studied, although it remains to be shown which is the optimal approach to disorders of smooth muscle motility. PMID- 11392629 TI - Neostriatal muscarinic receptor subtypes involved in the generation of tremulous jaw movements in rodents implications for cholinergic involvement in parkinsonism. AB - Several studies have shown that a number of pharmacological and neurochemical conditions in rats can induce jaw movements that are described as "vacuous" or "tremulous". For several years, there has been some debate about the clinical significance of various drug-induced oral motor syndromes. Nevertheless, considerable evidence now indicates that the non-directed, chewing-like movements induced by cholinomimetics have many of the characteristics of parkinsonian tremor. These movements are characterized largely by vertical deflections of the jaw, which occur in the same 3-7 Hz peak frequency that is typical of parkinsonian tremor. Cholinomimetic-induced tremulous jaw movements are suppressed by a number of different antiparkinsonian drugs, including scopolamine, benztropine, L-DOPA, apomorphine, bromocriptine, ropinirole, pergolide, amantadine, diphenhydramine and clozapine. A combination of anatomical and pharmacological research in rats has implicated M4 receptors in the ventrolateral neostriatum in the generation of tremulous jaw movements. Mice also show cholinomimetic-induced jaw movements, and M4 receptor knockout mice demonstrate subtantially reduced levels of jaw movement activity, as well as increased locomotion. Taken together, these data are consistent with the hypothesis that a centrally-acting M4 antagonist may be useful as a treatment for parkinsonian symptoms, including tremor. PMID- 11392630 TI - Facilitation of acetylcholine release and improvement in cognition by a selective M2 muscarinic antagonist, SCH 72788. AB - Current treatment of Alzheimer's Disease (AD) requires acetylcholinesterase inhibition to increase acetylcholine (ACh) concentrations in the synaptic cleft. Another mechanism by which ACh levels can be increased is blockade of presynaptic M2 muscarinic autoreceptors that regulate ACh release. An antagonist designed for this purpose must be highly selective for M2 receptors to avoid blocking postsynaptic M1 receptors, which mediate the cognitive effects of ACh. Structure activity studies of substituted methylpiperadines led to the synthesis of 4-[4 [1(S)-[4-[(1,3-benzodioxol-5-yl)sulfonyl]phenyl]ethyl]-3(R)-methyl-1-piperazinyl] 4-methyl-1-(propylsulfonyl)piperidine. This compound, SCH 72788, binds to cloned human M2 receptors expressed in CHO cells with an affinity of 0.5 nM, and its affinity at M1 receptors is 84-fold lower. SCH 72788 is a functional M2 antagonist that competitively inhibits the ability of the agonist oxotremorine-M to inhibit adenylyl cyclase activity. In an in vivo microdialysis paradigm, SCH 72788 increases ACh release from the striatum of conscious rats. The compound is also active in a rodent model of cognition, the young rat passive avoidance response paradigm. The effects of SCH 72788 suggest that M2 receptor antagonists may be useful for treating the cognitive decline observed in AD and other dementias. PMID- 11392631 TI - Pharmacodynamic profile of the M1 agonist talsaclidine in animals and man. AB - In functional pharmacological assays, talsaclidine has been described as a functionally preferential M1 agonist with full intrinsic activity, and less pronounced effects at M2- and M3 receptors. In accordance with this, cholinomimetic central activation measured in rabbits by EEG recordings occurred at a 10 fold lower dose than that inducing predominantly M3-mediated side effects. This pharmacological profile is also reflected in the clinical situation: Both in healthy volunteers and in Alzheimer patients--unlike after unspecific receptor stimulation through cholinesterase inhibitors--the mainly M3 mediated gastrointestinal effects (like nausea and vomiting) were not dose limiting. Rather, sweating and hypersalivation, mediated through muscarinic receptors, occurred dose-dependently and were finally dose-limiting. In contrast to talsaclidine, sabcomeline had a less pronounced functional M1 selectivity in pharmacological assays. This was also shown in anaesthetized guinea pigs where sabcomeline alone induced bronchoconstriction, and in the rabbit EEG where central activation and cholinergic side effects occurred in the same dose range. Neither drug, however, showed convincing improvement of cognitive functions in patients with mild-to-moderate Alzheimer's disease. This asks for a reassessment of the muscarinic hypothesis for the treatment of this disease. PMID- 11392632 TI - Exploring the potential for subtype-selective muscarinic agonists in glaucoma. AB - Pilocarpine has been used to lower intraocular pressure (IOP) in glaucoma patients for more than 100 years. Since the identification of five muscarinic receptor subtypes, there has been an interest in separating the IOP-lowering effects from the ocular side effects of pupil constriction and lens accommodation. However, all these actions seem to be mediated by the M3 receptor. A novel muscarinic receptor agonist, AGN 199170, that has no activity on the M3 subtype was compared to pilocarpine in a monkey glaucoma model. This compound lowered IOP suggesting that muscarinic agonists targeted at muscarinic receptors other than the M3 subtype may be able to selectively lower IOP. PMID- 11392634 TI - Left ventricular volume reduction for dilated cardiomyopathy. PMID- 11392633 TI - Elucidating the role of muscarinic receptors in psychosis. AB - Muscarinic receptors have been implicated in the regulation of cognition and psychosis based on pharmacological evidence from pre-clinical and clinical studies. Muscarinic agonists have shown promise in the clinic in improving cognition and reducing psychotic episodes in Alzheimer's patients. However, lack of selective muscarinic ligands has limited their use due to troublesome side effects observed at higher doses. Without selective ligands, it has been difficult to assign a specific muscarinic receptor subtype to these high order mental processes. Recent development of muscarinic receptor knockout mice has provided additional tools to investigate cognition and psychosis in behavioral assays and to determine the receptor subtypes associated with parasympathomimetic physiology. Biochemical studies indicate that the M1 receptor plays a significant role in regulating G alpha q-mediated signal transduction in the hippocampus and cortex. Behavioral studies suggest that the M4 receptor is involved in movement regulation and prepulse inhibition of the startle reflex, a measure of attention. These findings support a role for the development of M1 and M4 receptor agonists for diseases in which symptoms include cognitive impairment and psychotic behaviors. PMID- 11392635 TI - Benefits of beta-blockers in heart failure: a class specific effect? AB - There is now compelling evidence in favor of the use of beta-adrenergic antagonists for the treatment of chronic heart failure. In clinically stable patients who remain symptomatic despite the fact that they are already receiving an angiotensin-converting enzyme inhibitor, diuretics and digoxin, the addition of a beta-blocker has been shown to produce further improvements in cardiac function and structure as well as in the quality and quantity of life. However, although such benefits can be achieved with a number of beta-blockers, the relevant differences in the ability of inhibiting the adrenergic drive among the various agents in the same class could translate into quantitatively different clinical effects. At present, the question whether all beta-blockers confer equal benefit or not to heart failure patients remains unanswered, since only few studies have prospectively addressed the issue and overall evidence does not permit to draw a conclusion that one agent has to be preferred to another. A large ongoing trial, designed to compare the effects of metoprolol and carvedilol on all-cause mortality in chronic heart failure, will provide much of the information required. PMID- 11392636 TI - Ultrasonic myocardial tissue characterization: a methodological review. AB - Ultrasonic myocardial tissue characterization represents a relatively new diagnostic tool which allows integration of the conventional echocardiographic evaluation, in order to obtain specific textural parameters which reflect the myocardial ultrastructural texture. In particular, through this approach it is possible to obtain two different types of information: the first is static and consists of the absolute myocardial echo intensity that reflects the ultrastructural myocardial changes in different diseases; the second is dynamic and is related to the variations of echo intensity during the cardiac cycle which seem to be linked, even though not linearly, to the intrinsic myocardial contractility. Our research group has extensively applied this methodological approach to different pathophysiological models, in particular to essential hypertension. In the present review the technological evolution of the method and comparison with other research groups' experience with the specific pathophysiological models, are shown and discussed. PMID- 11392637 TI - Restrictive left ventricular filling pattern as a strong predictor of depressed baroreflex sensitivity in heart failure. AB - BACKGROUND: The aim of this study was to test the hypothesis that a restrictive left ventricular diastolic filling pattern, as an index of elevated pulmonary wedge pressure, would predict a depressed baroreflex sensitivity (BRS) in patients with chronic heart failure. METHODS: A total of 189 consecutive patients with an ejection fraction < or = 40% at echocardiography, in sinus rhythm and clinically stable for at least 1 month in oral therapy, underwent clinical examination, echo-Doppler study and the phenylephrine test. RESULTS: The correlations between the NYHA functional class, echo-Doppler variables and BRS were weak, although significant (r ranging from -0.15 to 0.40). However, patients with a deceleration time < 140 ms as an expression of restrictive filling, compared to those with a deceleration time > or = 140 ms, had a lower BRS (3 +/- 4 vs 6 +/- 4 ms/mmHg, p < 0.00001), a lower ejection fraction (20 +/- 6 vs 28 +/- 7%, p < 0.00001), greater left ventricular (end-diastolic volume index 137 +/- 43 vs 113 +/- 45 ml/m2, p < 0.00001) and left atrial dimensions (25 +/- 6 vs 20 +/- 5 cm2, p < 0.00001), more severe mitral regurgitation (3 +/- 1 vs 2.3 +/- 1, p < 0.00001) and were in a higher NYHA class (2.3 +/- 0.6 vs 1.8 +/- 0.5, p < 0.00001). Medications at the time of the study were similar in the two groups. At stepwise regression analysis, the deceleration time emerged as the most powerful independent predictor of a depressed BRS (< 3 ms/mmHg), followed by mitral regurgitation, age, and NYHA class (all data p = 0.0001). CONCLUSIONS: In patients with chronic heart failure, the presence of a restrictive left ventricular filling pattern is highly predictive of autonomic derangement as expressed by low values of BRS. PMID- 11392638 TI - Incidence and prevalence of ischemic heart disease in Italy: estimates from the MIAMOD method. AB - BACKGROUND: When considering large areas, population-based data on coronary events are generally lacking, and such is the case for the national level. While mortality data are currently and exhaustively collected, data regarding the incidence and prevalence are often available only for subgroups of the population. METHODS: The incidence and prevalence of coronary events were estimated using a mathematical method on the basis of official mortality and population data from national statistics and survival data on coronary events from the Area Friuli of the MONICA Project, and forecasted for northern, central and southern Italy. RESULTS: The incidence is described from 1970 to 1994 and projected to the year 2004; prevalence is reported at the years 1990 and 2000. The coronary event incidence has been decreasing since 1977 among men and since 1974, 4 years before the observed mortality decline, among women. The prevalence has continued to increase as a result of three main factors: increasing survival, population aging, and incidence trend. CONCLUSIONS: Incidence and prevalence data distributed for northern, central and southern Italy are essential to plan and implement major projects aimed at improving medical care services and to evaluate the impact of public health interventions and of spontaneously changing habits among the population. PMID- 11392639 TI - Potential influence of pre-infarction angina on myocardial viability and residual ischemia. AB - BACKGROUND: The effects of recent pre-infarction angina on myocardial viability and residual ischemia are unknown. This study evaluates them in 90 patients with a first Q-wave myocardial infarction using early dobutamine stress echocardiography. METHODS: Patients were classified according to the absence or presence of recent pre-infarction angina, defined as chest pain lasting < 30 min during a period of 7 days before the acute myocardial infarction. The infarct zone wall motion score index was calculated at baseline and at low- and peak-dose dobutamine stress echocardiography. All subjects underwent coronary angiography. RESULTS: Patients with unheralded myocardial infarction showed, in comparison with patients with recent pre-infarction angina, a significantly higher peak of creatine kinase serum levels (2630 +/- 1360 vs 1865 +/- 1562 IU/l, p = 0.002) and a higher number of leads with pathologic Q waves (3.2 +/- 1.3 vs 2.8 +/- 0.8, p = 0.002). The groups did not differ with regard to the infarct zone wall motion score index at rest (2.15 +/- 0.42 vs 2.18 +/- 0.31, p = 0.72) and at low- (1.86 +/- 0.52 vs 1.80 +/- 0.41, p = 0.55) and peak-dose (2.24 +/- 0.55 vs 2.19 +/- 0.58, p = 0.68) dobutamine stress echocardiography. The prevalence of myocardial viability (31 vs 48%, p = 0.15), homozonal (52 vs 58%, p = NS) or heterozonal ischemia (36 vs 23%, p = NS) was not statistically different between the groups of patients without and with recent pre-infarction angina. The angiographic patterns were similar. CONCLUSIONS: Recent pre-infarction angina is associated with a smaller infarct size but it does not seem to influence the ventricular contractile improvement or residual ischemia, detected at early dobutamine echocardiography, in patients with a first Q-wave myocardial infarction. PMID- 11392640 TI - "Hybrid" percutaneous and surgical coronary revascularization: selection criteria from a single-center experience. AB - BACKGROUND: The association of minimally invasive direct coronary artery bypass (MIDCAB) to percutaneous transluminal coronary angioplasty (PTCA) of large arteries with focal lesions can be an alternative therapeutic method for patients with multivessel coronary artery disease. We reviewed our experience regarding 42 patients treated at our Institute. METHODS: MIDCAB and PTCA of the circumflex or right coronary arteries > 3 mm were performed in 42 patients from September 1997 to December 1999. RESULTS: One patient died after MIDCAB in the operating room because of rupture of the left anterior descending anastomosis. Postoperative angiography confirmed patency of the internal mammary artery (IMA) graft in 92.3% of cases: 3 early IMA graft failures occurred. The success rate for PTCA was 98%: in 1 case the wire just would not cross a chronically and totally occluded right coronary artery. The in-hospital morbidity was 12.2%: 2 patients required urgent sternotomy respectively for cardiac tamponade and coronary artery bypass grafting on cardiopulmonary bypass. One patient developed atheroembolism after PTCA with recurrence of symptoms, progressive multiorgan failure and death. Two patients required PTCA on the IMA anastomosis because of early failure of the arterial graft. At a medium follow-up of 535 days, all 40 survivors are in Canadian Cardiovascular Society class I. CONCLUSIONS: Hybrid revascularization appears to be an effective treatment for selected patients with multivessel coronary artery disease. The immediate success seems related to the learning curve for MIDCAB. PMID- 11392641 TI - "Hybrid" myocardial revascularization: "the best of two worlds?" May be, but for a small number of selected patients. PMID- 11392642 TI - Ultrasound-assisted stent implantation in small size coronary arteries: a pilot study. AB - BACKGROUND: Many studies have indicated that a small lumen size is one of the most important predictors of acute events and of late restenosis after balloon angioplasty or stent implantation. In the last few years many studies have shown that intravascular ultrasound (IVUS) guidance makes it possible to optimize stent implantation. The aim of this pilot study was to evaluate the feasibility and safety of IVUS imaging of small vessels. Secondary endpoints were the immediate and long-term results of IVUS-guided elective BeStent implantation in small vessels. METHODS: Fourteen symptomatic patients with small coronary vessel (mean angiographic reference diameter 2.3 +/- 0.2 mm) disease underwent IVUS-guided BeStent implantation. IVUS success was defined as the achievement of a final minimal intrastent cross-sectional area (CSA) > 90% of the smaller reference lumen CSA. RESULTS: IVUS evaluation was feasible in all patients without any clinical or angiographic adverse events. Procedural success was achieved in all patients with implantation of a BeStent 15 mm. No major complication (death, myocardial infarction, stent acute or subacute thrombosis, coronary artery bypass, re-coronary angioplasty) occurred during the in-hospital phase. Two non flow-limiting, asymptomatic coronary dissections were detected after stent expansion. The post-stenting lesion stenosis rate decreased from 72.9 +/- 12.9% to 0.75 +/- 11.7% with an acute gain of 1.8 +/- 0.4 mm. The final IVUS minimal stent CSA was 5.6 +/- 1.1 mm2. The IVUS criteria of adequate stent expansion were reached in 11 (78.6%) patients. At 6 months of follow-up, the rate of angiographically diagnosed in-stent restenosis was 30.7%; the 6-month late loss in stent diameter was 1.1 +/- 0.6 mm. No patient died or presented with a myocardial infarction. The target lesion revascularization rate was 30.7%. CONCLUSIONS: Coronary IVUS-guided stenting can be performed in small vessels with a high success rate and low incidence of in-hospital complications. However, despite these encouraging short-term results, the long-term clinical and angiographic outcome is less favorable. Further larger and randomized IVUS studies, probably employing more aggressive IVUS criteria, are needed to clarify the true role of IVUS guidance in this particular field. PMID- 11392643 TI - Coronary artery bypass grafting in patients with dialysis-dependent renal failure: ten-year results. AB - BACKGROUND: Coronary artery disease has been reported to be accelerated in patients with chronic renal failure on maintenance dialysis. Coronary artery bypass grafting (CABG) in patients on long-term dialysis is still a debated issue. METHODS: We retrospectively reviewed 19 patients (12 men, 7 women, mean age 64 +/- 11.2 years) with end-stage renal disease who underwent CABG between 1990 and 2000. Operative procedures were CABG alone in 15 (78.9%) patients and CABG associated with valve procedures in 4 (21.1%) patients. RESULTS: The early (30-day) mortality rate was 10.5% (2 of 19 patients). Non-fatal complications occurred in 6 patients (31.5%). Four delayed deaths occurred; the actuarial survivals at 1, 2, 5 and 10 years were 0.86 +/- 0.14, 0.78 +/- 0.10, 0.68 +/- 0.13 and 0.54 +/- 0.15 respectively. Among 13 survivors the mean Canadian Cardiovascular Society class was 1.3 +/- 0.3 (p < 0.001 vs preoperatively). ANOVA procedures showed age (p = 0.01), Canadian Cardiovascular Society class > or = III (p < 0.001), urgent/emergency operation (p < 0.001), left ventricular ejection fraction < 0.50 (p < 0.001), a prior myocardial infarction (p = 0.01), a preoperative mean creatinine level > or = 5 mg/dl (p = 0.02) and a duration of dialysis > or = 60 months (p = 0.03) to be strongly related to early and delayed mortality. CONCLUSIONS: CABG in patients with dialysis-dependent chronic renal failure is associated with acceptable results. Accurate patient selection, early referral to surgery, and adequate perioperative management are advisable. PMID- 11392644 TI - Pulmonary atresia with ventricular septal defect: prevalence of deletion 22q11 in the different anatomic patterns. AB - BACKGROUND: Pulmonary atresia with ventricular septal defect (PA-VSD) is one of the most common cardiac defects associated with DiGeorge syndrome. The pattern of the pulmonary circulation determines the complexity of this type of heart disease. The aim of this study was to establish the prevalence of DiGeorge syndrome with deletion 22q11 in patients with simple and complex PA-VSD. METHODS: Since 1993 we have studied 128 consecutive patients affected by PA-VSD. In 90 of our patients the PA-VSD was considered "simple" (group I), because it was not associated with any other cardiac defects. In the other 38 children the PA-VSD was considered "complex" (group II) owing to the presence of heterotaxia, tricuspid atresia, a double-inlet left ventricle, transposition of the great arteries and congenitally corrected transposition of the great arteries. RESULTS: In group I, 38 patients (42%) had genetic syndromes or major extracardiac anomalies; deletion 22q11 was detected in 31% of cases. Major aortopulmonary collateral arteries were present in 50% of group I patients and in 57% of those with deletion 22q11. In group II, 10 patients (26%) had genetic syndromes or major extracardiac anomalies but none had deletion 22q11 (p < 0.005); in no case was the presence of major aortopulmonary collateral arteries observed (p < 0.005). CONCLUSIONS: PA-VSD is an anatomically and morphogenetically heterogeneous disease: in the setting of DiGeorge syndrome or velocardiofacial syndrome, PA-VSD is associated with a peculiar cardiac pattern and is due to deletion 22, whereas in case of nonsyndromic PA-VSD or when this disease is associated with different syndromes or with other types of cardiac defects, it is due to other morphogenetic mechanisms. PMID- 11392645 TI - The duration of atrial fibrillation influences the long-term efficacy of low energy internal cardioversion. AB - BACKGROUND: It is commonly held that long-lasting atrial fibrillation (AF), especially if associated with marked enlargement of the left atrium, is a negative predictive factor for both the recovery and the maintenance of sinus rhythm. The aim of the present study was to identify the clinical features of patients who have a greater likelihood of success both in the acute phase and, especially, in the medium-long term. METHODS: Since June 1997, we have performed low-energy internal cardioversion to 93 patients (66 males, 27 females, mean age 62 +/- 9 years, range 26-80 years) with a mean duration of AF of 922 +/- 1032 days. Seventy-four patients had heart disease and 19 isolated AF. External cardioversion had been previously performed in 79 patients to no avail. All patients underwent antiarrhythmic therapy and were followed for a period of 13 +/ 7 months. RESULTS: Low-energy internal cardioversion proved efficacious, restoring sinus rhythm, in 92% of patients (86/93) and inefficacious in 8% (7/93). In 24% (21/86) the procedure, although efficacious, was followed by early recurrence of AF which proved to be intractable in 52% (11/21). At the end of the session, 81% (75/93) of the patients maintained sinus rhythm. At the end of follow-up, 40% (38/93) maintained sinus rhythm. Of all the parameters considered in the two groups, the duration of AF was the only one which differed significantly between the group in sinus rhythm and that in AF, with regard to both the efficacy of the procedure in the acute phase (755 +/- 868 vs 1618 +/- 1359 days, p < 0.001) and the long-term outcome (655 +/- 5.8 vs 1107 +/- 1098 days, p < 0.05). CONCLUSIONS: AF lasting more than 2 years constitutes a negative predictive factor for both the recovery and the long-term maintenance of sinus rhythm. PMID- 11392646 TI - Coronary artery bypass grafts in a patient with isolated cardiac dextroversion. AB - Despite several controversies, the term "dextrocardia" usually defines a rare type of intrinsic cardiac abnormality due to a rotation disorder and resulting in a right-sided direction of the cardiac axis. According to the majority of experts, the extent of a dextrocardia associated with a situs solitus is termed "dextroversion". In such a rare condition, therefore, the relationships between the cardiac chambers and the other structures (that is superior and inferior venae cavae, liver, stomach) are modified whereas in case of dextrocardia with situs inversus, the relationships between the cardiac chambers and neighboring structures are preserved and the classical "mirror image" is shown. In 95% of cases with dextroversion, an associated cardiac abnormality has been described and, therefore, acquired heart diseases in patients with isolated dextroversion are extremely rare. To our knowledge, the present is the first case report describing a coronary artery bypass graft performed in a patient with isolated dextroversion. The technical aspects of the surgical procedure are also discussed. PMID- 11392647 TI - Double-outlet right ventricle with intact ventricular septum. AB - In the present case report, we describe a malformation consisting of a double outlet right ventricle with an intact ventricular septum diagnosed in a 3-day-old female newborn. To our knowledge 35 cases have been described in the literature. The diagnosis was made by two-dimensional echocardiography and confirmed by angiocardiography. An inadequate opening in the interatrial septum and hypoplasia of the mitral valve and left ventricle were present. In this condition the only outlet of the left ventricle is via the atrial septal defect. Balloon atrial septostomy was performed. At 1 month of age the patient required a right modified Blalock-Taussig shunt and surgical atrial septectomy. Five months after surgery, the patient was in good clinical conditions. PMID- 11392648 TI - Images in cardiovascular medicine. Descending thoracic aorta, infrarenal abdominal aorta and bilateral common iliac artery aneurysm: endovascular repair in only one session. PMID- 11392649 TI - State-of-the-art research in lower-limb prosthetic biomechanics-socket interface: a review. AB - Scientific studies have been conducted to quantify attributes that may be important in the creation of more functional and comfortable lower-limb prostheses. The prosthesis socket, a human-machine interface, has to be designed properly to achieve satisfactory load transmission, stability, and efficient control for mobility. The biomechanical understanding of the interaction between prosthetic socket and the residual limb is fundamental to such goals. The purpose of this paper is to review the recent research literature on socket biomechanics, including socket pressure measurement, friction-related phenomena and associated properties, computational modeling, and limb tissue responses to external mechanical loads and other physical conditions at the interface. There is no doubt that improved biomechanical understanding has advanced the science of socket fitting. However, the most recent advances in the understanding of stresses experienced at the residual limb have not yet led to enough clinical consensus that could fundamentally alter clinical practice. Efforts should be made to systematically identify the major discrepancies. Further research should be directed to address the critical controversies and the associated technical challenges. Developments should be guided to offer clinicians the quantification and visualization of the interaction between the residual limb and the prosthetic socket. An understanding of comfort and optimal load transfer as patterns of socket interface stress could culminate in socket design expert systems. PMID- 11392650 TI - Osseointegration in skeletal reconstruction and rehabilitation: a review. PMID- 11392651 TI - Intramedullary osseointegration: development of a rodent model and study of histology and neuropeptide changes around titanium implants. AB - A rodent model has been developed to explore intramedullary osseointegration and the phenomena of osseoperception. Osseointegration with endosseous titanium implants is frequently used in oral surgery. More recently, intramedullary osseointegration has been used for direct skeletal anchoring of amputation prostheses, a procedure that provides for a stable prosthesis with improved perception. Experimental, commercially pure titanium rods with threaded ends were surgically implanted in the intramedullary space of 18 rat femurs, and were left in place for 8 wk. Microscopic and immunohistochemical observation of the titanium/bone interface at this time-point indicated successful osseointegration with normal remodeled bone adjacent to the fixture. Calcitonin gene-related peptide activity was upregulated during the process of bone remodeling, and there was no significant inflammatory reaction. There was new, normal bone adjacent to and fully occupying the space between fixture threads. Innervation also appeared normal in remodeled bone, as indicated by immunohistochemical observation of small nerve fibers with the antibody Protein Gene Product 9.5 (PGP 9.5). The model will be used further to explore intramedullary osseointegration and osseoperception in connection to clinical applications. PMID- 11392652 TI - Long-term survival of regenerated cartilage on a large joint surface. AB - The one-year survival of regenerated cartilage on a large articular surface is presented using the McDowell in vivo model. The model provides a mechanically shielded environment in which regenerated cartilage can be protected from intra articular stresses while normal joint motion is maintained. New tissue was allowed to grow from bleeding subchondral bone for 12 weeks at which time the original mechanical environment was reintroduced. Our study showed that neo cartilage would grow to cover the entire joint surface of a patella and could survive for one year. Histologic observations indicated a maturing hyaline-like tissue. Biomechanical analyses showed that the regenerated cartilage became stiffer and less permeable within the time of this study. Biochemical evaluations demonstrated stable properties out to the longest time point. Control specimens, which were not shielded from stress, showed insignificant amounts of new tissue growing on the patellar surfaces. PMID- 11392653 TI - Measuring the effects of seating on people with profound and multiple disabilities--a preliminary study. AB - This paper describes a preliminary study to investigate a range of approaches that might be used for measuring the effects of special seating on people with profound and multiple disabilities and their carers. A number of tools are proposed for measuring the effects on quality of life, function and carer satisfaction. The results of applying these tools to measure the effects of intervention with customized molded seating on nine people with multiple disabilities are described. The results suggest that these tools are sensitive to this intervention, showing a general beneficial effect with good carer satisfaction. The study points the way towards application of these tools to people with a wider range of disabilities and to different interventions. PMID- 11392654 TI - Driving performance and workload assessment of drivers with tetraplegia: an adaptation evaluation framework. AB - The purpose of this study was to establish a baseline for further research on adaptation evaluation for drivers with disabilities. Driving performance and workload for 26 drivers with spinal cord injuries (tetraplegia) was studied and compared to a matched group of able-bodied drivers in a driving simulator. Drivers with tetraplegia used two types of hand-operated controls for accelerating and braking. Able- bodied drivers drove with standard pedals. The drivers with tetraplegia performed the driving task equally as well as the control group but had a slightly longer reaction time (10%). Workload assessment revealed that drivers with tetraplegia experienced a significantly greater time pressure and spent more effort than did the able-bodied drivers. They were also more tired from braking and accelerating. The drivers with tetraplegia using separate levers had greater standard deviation in lateral lane position (7 cm), while those using a combined lever were more tired from braking and accelerating. Observed differences could be interpreted as indicators of insufficient adaptation. PMID- 11392656 TI - Assistive technology to improve PC interaction for people with intention tremor. AB - Many patients with upper limb intention tremor encounter difficulties in mouse driven interaction with the personal computer (PC). An assistive technology system ("the Tremor Control System"), consisting of a motion-filtering software program that supports multiple interfaces, was developed and validated with 36 persons with Multiple Sclerosis in a multi-center trial. PC-tests, requiring basic functions such as cursor placement and click and drag function, were able to differentiate between patients and control subjects (ANOVA: p<0.05). A significant time improvement on the PC-tests was found when using an optimal alternative interface instead of the standard PC-mouse (paired t-tests: p<0.01 for Point & Click test, p<0.05 for Drag & Drop test and p<0.1 for Double Click test). A significant time improvement was found for the Double Click test (paired t-tests: p<0.05) when the motion-filtering program was implemented. The number of patients able to perform fully the PC-tests increased with the Tremor Control System. Patients with marked intention tremor seemed to profit especially from this assistive technology. These users reported that working with the Tremor Control System was less fatiguing and more comfortable compared to the use of the standard PC-mouse. PMID- 11392655 TI - Dietary and serum lipids in individuals with spinal cord injury living in the community. AB - A cross-sectional study of 189 community-dwelling persons with spinal cord injury (SCI) (a) assessed levels of dietary and serum lipids, (b) determined the proportion of persons whose levels were out of the recommended/desired range, and (c) identified predictors of dietary and serum lipids. Lipid levels were out of range for a substantial proportion of the sample. Older persons were likely to have higher serum cholesterol and higher triglyceride levels than younger persons. Men tended to have higher intake of dietary cholesterol and lower levels of HDL than women. Caucasians and Hispanic-Americans tended to have higher triglycerides than African-Americans. Persons who had lived with SCI less time tended to have higher saturated fat intake and higher triglycerides than those who had lived with it longer. Greater saturated fat intake was associated with higher serum cholesterol after controlling for age. Studies are needed that test the effectiveness of various interventions on controlling dietary and serum lipids for persons with SCI. PMID- 11392657 TI - A comparison of regular rehabilitation and regular rehabilitation with supported treadmill ambulation training for acute stroke patients. AB - The purpose of this pilot study was to compare differences in motor recovery between regular rehabilitation (REG), and regular rehabilitation with supported treadmill ambulation training (STAT) using the performance on a bicycle exercise test and the locomotor scale of the Functional Independence Measure (FIM-L). Twelve patients with acute strokes were randomly assigned to either REG or STAT for 2 to 3 weeks. The STAT group received daily gait training utilizing a treadmill with partial support of body weight. After intervention, the STAT group had higher oxygen consumption (11.34+/-0.88 vs 8.32+/-0.88 ml/kg/min, p=0.039), total workload (58.75+/-7.09 vs 45.42+/-7.09 watts, p=ns), and total time pedaling the bike (288.91+/-30.61 vs 211.42+/-30.61 s, p=ns) compared to the REG group. The FIM-L scores were not different for the two groups. This pilot study suggests that the STAT intervention is a promising technique for acute stroke rehabilitation, and that future studies with larger sample sizes are warranted to establish the effectiveness of this intervention. PMID- 11392658 TI - Vibratory-coded directional analysis: evaluation of a three-microphone/four vibrator DSP system. AB - A sound localization aid based on eyeglasses with three microphones and four vibrators was tested in a sound-treated acoustic test room and in an ordinary office. A digital signal-processing algorithm provided a determination of the source angle, which was transformed into eight vibrator codes each corresponding to a 45 degrees sector. The instrument was tested on nine deaf and three deaf blind individuals. The results show an average hit rate of about 80% in a sound treated room with 100% for the front 135 degrees sector. The results in a realistic communication situation in an ordinary office room were 70% correct based on single presentations and 95% correct when more realistic criteria for an adequate reaction were used. Ten of the twelve subjects were interested in participating in field tests using a planned miniaturized version. PMID- 11392659 TI - A three-microphone system for real-time directional analysis: toward a device for environmental monitoring in deaf-blind. AB - Sound localization is a problem for the hearing impaired, deaf, and deaf-blind. A laboratory prototype of a three-microphone digital system for real-time directional analysis of sound sources that is mounted on eyeglasses has been developed. A cross-correlation algorithm is used for determinations of time and phase differences between the three microphone signals. The equipment was evaluated in a sound-treated test room with noise and speech stimuli from 12 different directions. It has an adequate precision (within approximately +/- 10 degrees) for speech and noise bursts in the frequency range 0.5-4 kHz, and for testing with words in a sound field the error percentage is very close to 0 at 0 degrees azimuth. The robustness of the algorithm in terms of signal-to-noise ratio is documented and reaches about +8 dB in different directions for speech and up to 0 dB for noise bursts. It is concluded that the construction of a robust system for directional analysis with sufficient efficiency for further experiments has been achieved. PMID- 11392660 TI - Optimal circumference reduction of finger models for good prosthetic fit of a thimble-type prosthesis for distal finger amputations. AB - The prosthetic fit of a thimble-type esthetic silicone prosthesis was retrospectively reviewed in 29 patients who were fitted following distal finger amputations. The aim was to correlate prosthetic fit with the magnitudes of circumference reduction in the finger models used to produce the prostheses and to identify the optimum reduction for the best outcome. A good fit is achieved primarily by making the prosthesis circumferentially smaller than the segment of the residual finger (residuum) over which it "cups". The percentage reduction in circumference of the finger model against the residuum model was calculated by dividing the difference in circumference between the residuum model and the finger model by the residuum model circumference and multiplying the result by 100. The computed percentage circumference reduction in the finger models ranged from small (1-3), moderate (5-7), to large (8-9). Twelve of 15 patients whose finger models had between one to three circumference reductions had a loose prosthetic fit. Only two of 14 patients who had a larger model circumference reduction of between five to nine had loose-fitting prostheses. Two of five patients who had eight to nine model circumference reduction had an uncomfortably tight prosthetic fit. A 5-7% circumference reduction in the finger model was shown in this study to best translate into good fit of a thimble-type prosthesis for distal finger amputations. PMID- 11392661 TI - Stroke: who's counting what? AB - INTRODUCTION: Patients with stroke are often selected for epidemiological reporting and research using ICD-9-CM (ICD-9) diagnostic codes. This study addresses the accuracy of these codes in identifying patients with stroke. METHODS: A sample of 279 patients with new stroke and 392 non-stroke (no evidence of new stroke) patients were identified by medical record review from 11 Veterans Affairs Medical Centers. Administrative records containing ICD-9-CM diagnoses were matched with this sample. Coding sensitivity and specificity were determined using individual ICD-9 codes and two coding algorithms. RESULTS: Significant variation was found in the accuracy of cerebrovascular ICD-9-CM codes in identifying patients diagnosed with stroke. Two coding algorithms were identified with the following performance statistics based on the sampled populations: 1) 91 percent sensitivity, 40-percent specificity; and 2) 54-percent sensitivity, 87 percent specificity. DISCUSSION/CONCLUSIONS: Variability in identifying patients with stroke using ICD-9 codes has been reported in the literature and confirmed. Two coding algorithms for maximizing sensitivity or specificity are proposed. Caution is urged when using ICD-9-coded administrative data to identify patients with stroke. PMID- 11392662 TI - Eugene F. Murphy, PhD, and early VA research in prosthetics and sensory aids. PMID- 11392663 TI - Organisation of the nervous system in the Acoela: an immunocytochemical study. AB - In order to broaden the information about the organisation of the nervous system in taxon Acoela, an immunocytochemical study of an undetermined Acoela from Cape Kartesh, Faerlea glomerata, Avagina incola and Paraphanostoma crassum has been performed. Antibodies to 5-HT and the native flatworm neuropeptide GYIRFamide were used. As in earlier studies, the pattern of 5-HT immunoreactivity revealed an anterior structure composed mainly of commissures, a so-called commissural brain. Three types of brain shapes were observed. No regular orthogon was visualised. GYIRFamide immunoreactive cell clusters were observed peripherally to the 5-HT immunoreactive commissural brain. Staining with anti-GYIRFamide revealed more nerve processes than did staining with anti-FMRFamide. As no synapomorphies were found in the organisation of the nervous system of the Acoela and that of the Platyhelminthes, the results support the view that the Acoela is not a member of the Platyhelminthes. PMID- 11392664 TI - Fine structure of the retino-optic nerve junction in the chicken. AB - The aim of this study is to investigate a fine structure of the retino-optic nerve junction in the chicken. We especially focused on the myelin sheaths and astrocytes in the intraocular optic nerve (ION) and its adjoining parts. A part of the axons of retinal nerve fiber layer (NFL) were myelinated. Ganglion cell axons were ensheathed by loose myelin in the NFL and by a compact one in the ION and optic nerve (ON). Myelin structure changed from loose type to a compact one within the very narrow NFL-ION junction. Loose myelin forming cells are dark type of oligodendrocytes in the retina. From the most peripheral ON to the choroidal part of ION, astrocytes contained abundant microtubules. The optic nerve around the lamina cribrosa is exposed to mechanical force during eye movement. It is suggested that these microtubules may perform the cytoskeletal function. Astrocytes in the retinal part of ION had longer processes filled with abundant gliofilaments. They may provide the mechanical support for the ganglion cell axons, which are exposed directly to intraocular pressure. Although astrocytes in the retinal level of ION extended their processes into the retina, their soma was never found in the retina. PMID- 11392665 TI - Protein kinase C activity contributes to endothelial hyperpermeability during early angiogenesis in the chick chorioallantoic membrane. AB - During angiogenesis in the chick chorioallantoic membrane (CAM), microvascular proliferation continues through day 12 of the 18-day CAM lifespan. Up to day 4.5, the neovascularization is associated with endothelial hyperpermeability and differentiation of restrictive barrier function occurs abruptly at day 5.0. Although exogenous activation of cAMP/protein kinase A (PKA) signaling served to decrease macromolecular extravasation at day 4.5, endogenous signaling cascades responsible for the temporal hyperpermeability remain uncertain. Here, we evaluated protein kinase C (PKC) function in the CAM endothelium at day 4.5 and day 5.0. The specific, broad-based PKC inhibitor calphostin C reduced basal levels of FITC-dextran 40 extravasation at day 4.5. Bisindolymaleimide (BIM), which inhibits selective PKC isoforms, also reduced temporal FITC-dextran 40 efflux, but to a lesser extent than calphostin C. Activation of PKC activity by phorbol-12, 13-didecanoate (PDD) or phorbol-12, 13-dibutyrate (PDBu) at day 5.0 served to partially de-differentiate barrier properties of the angiogenic endothelium. The associated elevation of FITC-dextran 40 extravasation occurred without interendothelial gap formation along the junctional clefts. Together, these results are consistent with the interpretation that PKC activity contributes, in part, to CAM endothelial hyperpermeability at day 4.5. Furthermore, down-regulation of PKC signaling correlates temporally with the ontogeny of restrictive barrier function at day 5.0. PMID- 11392666 TI - Distribution of reticulospinal neurons in the chicken by retrograde transport of WGA-HRP. AB - To determine the distribution of reticulospinal (RS) neurons in the chicken, WGA HRP was injected into the cervical or lumbosacral enlargement either unilaterally or bilaterally. The brainstem reticular nuclei sent largely descending fibers to both the spinal enlargements. The mesencephalon (medial and lateral mesencephalic reticular formation) and the rostral pons (nucleus reticularis [n.r.] pontis oralis) project mainly to the cervical enlargement. RS neurons were mainly distributed from the pontomedullary junction to the rostral medulla including n. r. pontis caudalis and pars gigantocellularis, n. r. gigantocellularis, n. r. parvocellularis, n. r. paragigantocellularis, and n. r. subtrigeminalis. It is suggested that the majority of these neurons send axons at least as far as the lumbosacral enlargement. In the lower medulla, RS neurons were distributed in the dorsal and ventral parts of the central nucleus of the medulla. PMID- 11392667 TI - Permeability barrier in the mantle epithelium lining the testis in the apple snail Pomacea canaliculata (Gastropoda: Ampullariidae). AB - Intercellular junctions are studied in the epithelium lining the testis of the freshwater snail Pomacea canaliculata by conventional staining and lanthanum tracer techniques. The junctional complex consists of belt desmosomes and septate junctions. Septate junctions are of the pleated-sheet type and they are constantly associated with mitochondria. Gap and tight junctions appear to be absent. These septate junctions seem to be the structural correlate of an epithelial permeability barrier that separate the testis from the extrapallial space where the shell elements are deposited. These junctions may contribute to a functional barrier in the male gonad of Pomacea canaliculata. The results indicate that freshwater prosobranchs have junctional structures very close to those found in other molluscs. PMID- 11392668 TI - Role of water-soluble matrix fraction, extracted from the nacre of Pinctada maxima, in the regulation of cell activity in abalone mantle cell culture (Haliotis tuberculata). AB - In mollusks, the mantle is responsible for the secretion of an organic matrix that mineralizes to form the shell. A model of mantle cell culture has been established from the nacreous gastropod Haliotis tuberculata. First, viability of cells, quantified by the MTT (3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide) reduction assay, was monitored in order to determine a cell density and a time-culturing period in order to investigate biomineralization processes in vitro. During the first 11 days of culture, an increase of MTT response demonstrated an activation of cultured cells mitochondrial activity as confirmed by the total protein content assay. The effect of a water-soluble extract from the organic matrix of Pinctada maxima (WSM) was tested on this cell culture system for 11 days-period exposure. WSM reduced the global viability of mantle cells in a dose-dependent way which corresponded to a cell death. Alkaline phosphatase activity normalized to total protein content increased in the presence of WSM. This increase may be due to an activation of cells and a selection of one (or a few) cell type(s). Further investigations will help us to determine this selectivity issue. PMID- 11392669 TI - Apoptosis and epithelial phagocytosis in mitomycin C-treated human pulmonary adenocarcinoma A549 cells. AB - Fate of human cancer cells damaged by mitomycin C was investigated by electron microscopy. In a monolayer culture of pulmonary adenocarcinoma A549 cells, the antitumor drug mitomycin C induced dilation of the smooth endoplasmic reticulum and Golgi apparatus. Simultaneously, the myelin figures, autophagosomes and dense tonofilament bundles were formed in the cytosol. Nuclear changes also included nucleolar condensation, as well as the disappearance of the karyosomes and thinned marginal heterochromatins. These nuclei came to be rigid and densely chromatic, finally resulting in apoptosis. There was no alteration in the mitochondria or rough endoplasmic reticulum. Meanwhile, some remaining intact A549 cells extended their cytoplasm toward detached cells and engulfed them. These results indicate that mitomycin C induces apoptosis in A549 cells and concurrently stimulates epithelial phagocytotic activity against their own damaged cells. PMID- 11392671 TI - Cytological effects of experimental exposure to Hg on the gill epithelium of the European flat oyster Ostrea edulis: ultrastructural and quantitative changes related to bioaccumulation. AB - The significance of ultrastructural changes in the gill epithelial cells as a parameter of detection of Hg exposure in the flat oyster, Ostrea edulis, was tested by a 34-day exposure to 5 microg l(-1) of Hg. The concentration of Hg (38.76 microg g(-1) dry weight) in gill tissue was maximal after 25 days and then decreased. The histological pattern of gill filaments in control samples did not vary throughout the experiment, except for the volume of mucocytes after 4 days of exposure, as an adaptation to experimental conditions. This volume increased significantly and then decreased according to the accumulation of Hg in the gills. After 18 days, absorptive and ciliated cells of the gill epithelium showed blebs in microvilli membranes, discocilia and swollen mitochondrial cristae. Both cell types showed distinct cellular lysis stages after 25 days of exposure. These are the target cells of Hg and other metals and the reported hypertrophy of mucocytes increase occurs in response to pollution by Hg, which could contribute to the detoxification process. PMID- 11392670 TI - Effects of continuous and intermittent exposure of lactating mothers to aroclor 1242 on testicular steroidogenic function in the adult male offspring. AB - Polychlorinated biphenyls (PCBs) are worldwide pollutants and have caused hazardous effects on many animal species including humans. They have been detected in human milk and therefore exposure of newborns to PCBs is unavoidable if they are breast-fed. We present our findings on two experiments performed to test the effects of intermittent and continuous exposure of lactating rats to two different doses (80 microg and 8 microg) of Aroclor 1242 (a PCB congener) on testicular steroidogenic function of their adult male offspring. In experiment I, three groups of lactating dams received daily subcutaneous (SC) injections of either corn oil, 80 microg of Aroclor 1242 and 8 microg of Aroclor 1242 in corn oil, respectively. In experiment II, three groups of lactating dams received two SC injections per week of either corn oil or Aroclor 1242 (80 microg and 8 microg) in corn oil, respectively. Pups in all groups (n=8 per group) were weaned at day 21 and were raised on a normal diet until sacrificed at 90 days. Experiment I: Leydig cell number per testis was significantly (P<0.05) increased and the average volume of a Leydig cell was significantly (P<0.05) reduced in both groups of Aroclor-exposed rats compared to corn oil controls. Both doses of Aroclor resulted in reduced (P<0.05) serum testosterone levels compared to corn oil-treated controls. LH-stimulated testosterone production per testis and per Leydig cell was lower in Aroclor-exposed rats compared to controls. Experiment II: No changes were observed in Leydig cell size and number per testis among the three groups. Serum LH, testosterone and LH-stimulated testicular testosterone production in offspring rats of Aroclor-treated dams were not significantly different (P>.05) from the offspring of corn oil-treated dams. However, these parameters were lower in value in the offspring of dams treated with Aroclor 80 microg compared to the other two groups. LH-stimulated testosterone secretory capacity per Leydig cell was significantly lower in offspring of dams treated with Aroclor compared to controls. Serum T4 and T3 levels were not significantly different among the Aroclor-exposed and control rats in both experiments. These results demonstrate that continuous exposure of lactating mothers to 8 and 80 microg of Aroclor 1242 causes hypotrophy and malfunctioning of Leydig cells in the adult male offspring resulting in a hypoandrogenic status. Intermittent treatment of lactating mothers with 80 microg of Aroclor (but not with 8 microg of Aroclor) also produced malfunctioning of Leydig cells and a hypoandrogenic status in the absence of Leydig cell hypotrophy. However, the Aroclor 8 microg dose was ineffective to produce the above effects. PMID- 11392672 TI - Immunohistochemical and morphological studies on the human fetal cochlea: a comparative view on methods. AB - The preservation of morphology and antigenicity can vary uncontrollably with human fetuses since these rely heavily on immediate fixation of the temporal bone following spontaneous abortion. Once good fixation is established, there is the question of the approach taken for morphologic and immunohistochemical studies. To achieve maximal preservation for the purpose of studying normal and pathologic fetal cochleae, commonly used preparation methods for analyzing the cochlea were reviewed and compared for both immunohistochemical and morphologic studies. Cochleae obtained after spontaneous abortion ranged from the 9th gestational week to birth. Four different methods were compared for morphologic study: the block surface method; a microslicing technique; paraffin; and celloidin sectioning. For immunohistochemical study, three methods were compared: pre-embedding; paraffin; and frozen sectioning. For morphologic preservation, the block surface method gave best overall results, showing good representation of the fetal cochlea for surface preparation, light, and electron microscopy. Celloidin sectioning was also found to show good light microscopic results for both the middle and inner ear. To achieve optimal results, preservation quality, fixation procedures, and antibody all contribute to the efficacy of a methods choice. PMID- 11392673 TI - Effect of the GnRH-agonist leuprolide on colonization of recipient testes by donor spermatogonial stem cells after transplantation in mice. AB - Gonadotropin-releasing hormone (GnRH)-agonist or antagonist treatment supports recovery of spermatogenesis after irradiation damage in the rat and appears to be beneficial to colonization of recipient testes after spermatogonial transplantation from fertile donors to the testes of infertile recipients in rats and mice. In the present study, we quantified the effect of treatment of recipient mice with the GnRH-agonist leuprolide acetate on the extent of colonization by donor spermatogonial stem cells in the recipient testis. Testis cells from mice carrying transgenes, which produce beta-galactosidase in spermatogenic cells, were used as donor cells for transplantation to allow for quantification of donor spermatogenesis in the recipient testis by staining for enzyme activity. Donor cell colonization 3 months after transplantation was compared between recipients receiving leuprolide in different treatment protocols and untreated control mice. Two injections of leuprolide 4 weeks apart prior to transplantation with as little as 3.8 mg/kg resulted in a pronounced improvement in the number of donor-derived spermatogenic colonies as well as in the in the area of recipient seminiferous tubules occupied by donor cell spermatogenesis. Improved colonization efficiency by treatment with GnRH-agonist can make the technique of spermatogonial transplantation applicable to situations when only low numbers of donor cells are available. PMID- 11392674 TI - A modified cryosection method for mouse testis tissue. AB - We have previously described a modified cryosection technique that improved the quality of histological sections as well as increasing the sensitivity to detect specific mRNA expression during early insect embryogenesis. Here, we report the use of this technique to produce high-quality sections of mouse testis tissue. The possibility of applying this technique to section the loose rat testis tissue is also discussed. PMID- 11392675 TI - Compliance with antiretroviral regimens. PMID- 11392676 TI - Medical-legal issues in HIV treatment: Part 1. PMID- 11392677 TI - Influence of psychiatric diagnoses and symptoms on HIV risk behavior in adults with serious mental illness. AB - HIV seroprevalence among people with serious mental illness has increased substantially in recent years. Although the prevalence of HIV behavioral risk factors has been well documented, few researchers have chosen to investigate the impact that psychopathology may have on HIV risk behavior. A review of the literature on psychiatric diagnoses related to HIV sexual risk reveals that bipolar disorder and perhaps schizophrenia are related to increased HIV risk. Further, persons in whom both Axis I and Axis II disorders are diagnosed appear to be at greater HIV behavioral risk. Research on psychiatric symptoms shows that illness severity may vary in its relation to HIV risk according to the specific symptoms present and that excited and possibly positive symptoms may be predictors of HIV sexual risk behavior. However, there is a need for research to elucidate these relationships. Future efforts that will add most to the literature will include examinations of psychiatric symptoms and illness severity, adequate sample sizes, statistical tests showing prediction of HIV risk behavior rather than the correlation with it, controls for substance abuse or dependence, and comparisons of psychopathology across HIV-positive and HIV negative persons. PMID- 11392678 TI - Improving immune function and controlling viral replication in HIV-1-infected patients with immune-based therapies. AB - Restoring and preserving immune function is a key component to successfully managing HIV-1 disease. Phase II/III studies have evaluated the safety and immunologic effects of immune-based therapies, including granulocyte-macrophage colony-stimulating factor, interleukin-2, and an inactivated HIV-1 immunogen, as adjuncts to antiretroviral therapy. Addition of each of these immune-based therapies to a background antiretroviral regimen enhanced, to varying degrees, immunologic function and suppression of viral replication in HIV-1-infected patients, suggesting a potential role for immune-based therapies in the treatment of HIV-1 disease. Further studies are needed to better characterize specific immunologic and virologic effects in different patient populations and to determine their impact on clinical outcomes. PMID- 11392679 TI - A near-fatal hypersensitivity reaction to abacavir: case report and literature review. AB - A near-fatal hypersensitivity reaction to abacavir developed in a 62-year-old HIV seropositive man who had been sensitized 17 months before presentation. Six days after he was rechallenged, acute respiratory distress developed, requiring mechanical ventilation for 2 weeks. Four days after extubation, he was again rechallenged. Hours later, the patient experienced anaphylactic shock, requiring mechanical ventilation for 3 weeks, aggressive volume resuscitation, and vasopressor support. Recovery was complicated by acute tubular necrosis, digital necrosis, and a GI bleed. This report reviews the mechanisms of action, efficacy, and adverse reactions of abacavir and illustrates the danger of serially rechallenging patients with this agent. PMID- 11392680 TI - [Evaluation of polymerase chain reaction method for monitoring of fungal infection in compromised hosts]. AB - Diagnosis of fungal infections in compromised hosts has been difficult because of insufficient sensitivity and specificity of conventional methods such as culturing and serum testing. Therefore, antifungal agents are usually started in febrile patients who are resistant to antibiotics even if these monitoring tests were negative. In this study, therefore, in order to increase the reliability of these monitoring, polymerase chain reaction (PCR) methods for detection of blood fungus were also performed in compromised hosts including 14 patients with hematological malignancies and one with solid tumor who were undergoing chemotherapies. From these patients, total of 56 peripheral blood samples was collected periodically, irrespective of the presence of infectious signs. At each time point of venopuncture, status of the patient was allocated to one of the followings: A, receiving an intravenous antifungal therapy because of sustaining fever which had not responded to prior antibiotic therapies and also positive for culturing and/or serum beta-D-glucan tests; B, receiving an additional intravenous antifungal therapy but negative for culturing and serum-tests; C, febrile but not yet receiving any intravenous fungal therapy; D, afebrile status. During the study, 10 blood samples from 3 patients were allocated in group A, and one sample of them was positive while remaining 9 were all negative for PCR. Six samples from 4 patients were in group B, and one was PCR positive while remaining 5 were negative. Fifteen samples from 7 patients were in group C, and 3 were positive and 12 were negative for PCR. Twenty-five samples were in group D, and 5 were positive and 20 were negative for PCR. Thus, the results from fungal PCR in these patients were in some case showed discrepancies from those expected from the clinical course and/or conventional monitoring tests. Further evaluation of fungal PCR may gain insight into the more precise diagnosis of fungal infection in these patients. PMID- 11392681 TI - [Genetic analysis of biosynthesis of polyketide compound in Streptomyces avermitilis]. PMID- 11392682 TI - [Study of the novel compounds modulating signal transduction system from microbial origin]. PMID- 11392683 TI - [Molecular mechanize and clinical feature of azole-resistant Candida albicans strains and new strategy of treatment]. PMID- 11392684 TI - [Investigation of msrSA gene coding for resistance to macrolide antibiotics by active efflux and its molecular epidemiology]. PMID- 11392685 TI - Cost-effectiveness of HIV therapies in different economic worlds. PMID- 11392687 TI - VaxGen AIDS vaccine clears phase III safety hurdle again. PMID- 11392686 TI - Behavioral interventions and host mucosal factors in transmission of HIV. PMID- 11392688 TI - Hospitals switch to safer needles: advocates push to avoid injuries. PMID- 11392689 TI - African AIDS "not caused by more virulent strain". PMID- 11392690 TI - Choosing which nuke to use first. PMID- 11392691 TI - Persistent herpes simplex virus infection. PMID- 11392692 TI - New HIV vaccine strategy shows promise in monkeys. PMID- 11392693 TI - Policy watching. PMID- 11392694 TI - HAART is cost-effective and improves outcomes. AB - An agenda for health care reform is well beyond the scope of this column. However, the agenda for providers of HIV care in managed care settings is clear. On a programmatic level, we know many of the solutions already. For example, there are published data supporting the role of experienced providers for HIV care. Outcomes are improved when patients with HIV receive care from "experts" (i.e., providers with experience in managing the disease). HAART improves outcomes; patients live longer and have more sustained viral load suppression, and costs of care are lower. Adherence to HAART is also beneficial and results in the same improved outcomes. The use of newer technologies to select these expensive antiretroviral regimens also results in these improved outcomes. Early treatment to which patients are adherent, access to experienced HIV providers, and a multidisciplinary program that delivers that care and the needed wraparound services make sense. In the era of HAART, it is often said that HIV is a chronic, manageable illness. Unfortunately, this applies to only a segment of the HIV infected population. The findings of Freedberg and Bozzette and their colleagues continue to support a model of well-timed, well-coordinated HIV care. While this model of care is expensive, it is less expensive than inpatient care or the resultant poorer outcomes in the underserved, in nonadherent patients, or in those whose care is delivered by less experienced providers. There is no price to be placed on improved quality of life. PMID- 11392695 TI - HIV and cardiovascular risk factors. AB - There is growing concern that the metabolic complications associated with HIV and antiretroviral therapy may lead to accelerated coronary artery disease (CAD). Traditional cardiovascular risk factors, such as hypertension, hyperlipidemia, diabetes mellitus, and visceral fat accumulation, are increasingly seen in HIV patients receiving HAART. These factors are in addition to nonreversible risk factors, such as male sex, age greater than 40 years, and family history of premature CAD. Patients may also be smokers and may have a sedentary lifestyle, both of which predispose to significant CAD. In older patients and those with other risk factors, there may be an accentuation of these risk factors by HAART, although these factors also occur in young patients with no other risk factors. It is still unknown whether the factors predispose HIV patients to accelerated cardiovascular disease. Short-term studies, including 2 large cross-sectional studies, do not show an increased risk of cardiovascular complications or cardiac death, but longer follow-up is needed to answer such questions effectively. Even if patients are not at increased risk for cardiovascular disease, they are at least at the same risk as HIV-negative, age-matched persons with similar risk factors. It is, therefore, pertinent to identify and effectively manage those risk factors that can be modified. PMID- 11392696 TI - Risk of perinatal transmission with treatment combinations of intrapartum and newborn zidovudine monotherapy. AB - OBJECTIVE: To determine perinatal transmission rates when zidovudine (ZDV) was given either to mothers after labor had begun, to their infants postnatally, or both. METHODS: Woman-infant pairs who received intrapartum and newborn ZDV therapy between January 1, 1992, and July 1, 1998, were considered. A medical record review identified female subjects known to be HIV-infected. All women who were given treatment with any antiretroviral drug before the onset of labor were excluded. All women satisfying eligibility criteria were enrolled. Nonparametric tests were used for analysis. The transmission rate of the study population was compared with rates reported for mother-infant pairs in the United States who did not receive ZDV. RESULTS: Administration of ZDV therapy after onset of labor resulted in a transmission rate of 11.9% (n = 59; 95% confidence interval [CI], 4.9 to 22.9). The overall perinatal transmission rate among women who received ZDV therapy after the onset of labor among sites in New York City was 6.3% (n = 32; CI, 0.8 to 20.8) compared with 18.5% (n = 27; CI, 6.3 to 38.0) among other sites. Administration of intrapartum therapy alone (n = 9), intrapartum plus newborn therapy (n = 37), and newborn therapy alone (n = 13) resulted in transmission rates of 11.1% (CI, 0.3 to 48.2), 13.5% (CI, 4.5 to 28.8), and 7.7% (CI, 0.2 to 36.0), respectively. CONCLUSION: The transmission rates reported here are lower than rates reported when antiretroviral therapy was not administered. PMID- 11392697 TI - Factitious HIV syndrome in young women. AB - Factitious HIV infection has been observed at our center in women presenting with a false history of HIV/AIDS. In a 2-year period, 4 women presented for HIV related care, indicating they were HIV-seropositive, while repeated serologic testing revealed no evidence of HIV infection. In all cases, the women were either quite angry or appeared surprised when told that they did not have HIV infection. A common denominator in all 4 women was a history of prolonged sexual, physical, or emotional abuse. Three of the 4 had been to other physicians, changing doctors as soon as the absence of HIV infection was established. Appropriate psychiatric support is an important aspect in care of these women, although it may not be accepted. All presentations of HIV infection should be confirmed either by identifying hard-copy data of HIV test results or by retesting all patients before evaluation and treatment of presumed HIV-related illnesses. PMID- 11392699 TI - Systems thinking: managing the pieces as part of the whole. PMID- 11392698 TI - Making it work: planning and executing a successful LIS installation. AB - This article relates how to plan and execute a successful laboratory information system (LIS) installation for those installing a first LIS, converting to a new LIS, or upgrading the current LIS. It covers the planning process, schedule adherence, vendor relationships, project team management, and system readiness, as well as post-installation issues. PMID- 11392700 TI - Successful community-based laboratory services program for long-term care facilities, Part 1. AB - Hagerstown Medical Laboratory, Inc. (HML) is a regional reference laboratory in Hagerstown, Maryland, that provides laboratory services to more than 50 long-term care facilities (LTCFs, or nursing homes) in Western Maryland and West Virginia. HML also operates the rapid response laboratory at Washington County Hospital and performs house calls for homebound or bedridden patients through its Nursing Home Program (NHP). This article relates HML's successful experience with an outreach program that provides laboratory services to LTCFs. PMID- 11392701 TI - It's about time. AB - Managers need more time. They juggle five tasks at once. As they work on one, the other four fall to the ground. They make a list of 10 things to do and spend all day working on number 11. Yes, managers need more time, but they already have all the time there is. So they must make better use of the time they have. Many articles address this issue. Managers read them all, but none of them work. This one will. PMID- 11392702 TI - Clinical laboratory health screening tests: a regulatory case study. PMID- 11392703 TI - An uncomfortable issue: dealing with substance abuse. PMID- 11392704 TI - Protecting against the disgruntled employee. PMID- 11392705 TI - Dollar$ & $en$e. Part VI: Knowledge management: the state of the art. AB - In Part I of this series, I introduced the concept of memes (1). Memes are ideas or concepts--the information world equivalent of genes. The goal of this series of articles is to infect you with memes so you will assimilate, translate, and express them. No matter what our area of expertise or "-ology," we all are in the information business. Our goal is to be in the wisdom business. In the previous articles in this series, I showed that when we convert raw data into wisdom, we are moving along a value chain. Each step in the chain adds a different amount of value to the final product: timely, relevant, accurate, and precise knowledge that then can be applied to create the ultimate product in the value chain- wisdom. In part II of this series, I introduced a set of memes for measuring the cost of adding value (2). In part III of this series, I presented a new set of memes for measuring the added value of knowledge, i.e., intellectual capital (3). In part IV of this series, I discussed practical knowledge management tools for measuring the value of people, structural, and customer capital (4). In part V of this series, I applied intellectual capital and knowledge management concepts at the individual level, to help answer a fundamental question: what is my added value (5)? In the final part of this series, I will review the state of intellectual capital and knowledge management development to date and outline the direction of current knowledge management initiatives and research projects. PMID- 11392706 TI - What are you doing to help professionally develop your staff? PMID- 11392707 TI - How to help your staff strengthen customer service: a do-able approach. AB - In today's high-pressure, speed-oriented environment, health-care managers face the challenge of helping their staff achieve a higher level of customer service without overwhelming them. Although raising the bar on customer service is crucial to stay competitive, if managers expect staff to institute many customer service improvements at once, staff often respond with confusion, frustration, and resistance. Then results are disappointing, and managers flounder in their efforts to inspire staff to stretch toward higher standards. By prioritizing customer service goals and selecting one goal at a time as the focus, you and your team can institute and sustain improvements successfully. Whether the goal is improving "meeting and greeting," response time, handling complaints more effectively, providing better instructions, or other customer service goals, the one-goal-at-a-time focus makes it possible for staff to understand the desired change, embrace it, learn it, practice it, and savor positive results. By following a simple 6-step process and applying this same process to one goal after another, staff become familiar with the process and it becomes easier to apply in pursuit of future goals. The result: a higher return on the investment of your efforts and your staff's precious time and attention. PMID- 11392708 TI - Implementing an outreach program. AB - Outreach programs are providing solutions to the problems of finding new revenue sources and dealing with under-use of equipment. What better way to increase monies than to attract new clients? How much more efficient can you be than to ensure that your instrumentation is used to capacity? Before you hang up your banner and declare yourself open for outreach business, ask yourself a few questions, suggested by those who are doing outreach and have prioritized the necessities for success: Can we geographically make this work (turnaround time, courier systems)? Will others want to partner with us (Are we reputable? Do we add value to our service?)? Will our administration support this effort (funds and manpower, as well as verbal endorsement)? We have the machinery--do we have the staff to do this? Is our staff customer-service oriented? Are we prepared to handle additional compliance/billing issues? In short, you need a plan. This Template Topic offers a checklist for the essential components of outreach programs, allowing you to inventory your readiness for such an endeavor or, if you already are involved in outreach, to take stock of your current situation and acknowledge areas for improvement. Pay particular attention to staff concerns. Those on the front line, interacting with patients/clients, can make or break an outreach program. This Template Topic will provide the support you need as you reach out to new initiatives and opportunities to strengthen your business and your importance to your medical community. PMID- 11392710 TI - Are we giving away the farm? PMID- 11392709 TI - IOM report on medical errors: CLMA members speak out. PMID- 11392711 TI - In need of assistance. Widening quarterly losses a common theme in assisted living industry. PMID- 11392712 TI - Two systems, two moves. Big Apple players tinker with their governance. PMID- 11392713 TI - Slow to change. Hospital boards insulated from industry reshaping. PMID- 11392714 TI - GPO maverick. MedAssets now a Georgia peach in feisty field of hospital cooperatives. PMID- 11392715 TI - Island fever. Santa Catalina's tiny hospital still struggling, but new CEO has big plans. PMID- 11392716 TI - A second opinion. Study rebuts Calif. docs' contention they're underpaid and in short supply. PMID- 11392717 TI - Quite quarrelsome. Provider advocates still argue against privacy rules now in effect. PMID- 11392718 TI - Dear Tommy ... lawmakers' letter proposes fixes for Medicare. PMID- 11392719 TI - The ways of the White House. PMID- 11392720 TI - A sharper image. Father of the MRI honored for invention that forever changed diagnostics. PMID- 11392721 TI - Ticker shock. Stock market's volatility shows up on first-quarter earnings reports. PMID- 11392722 TI - Budgets may get bloodied. American Red Cross plans to raise average price of blood by nearly one-third. PMID- 11392723 TI - Prescription drug talk cooled off faster than a Texas thunderstorm. PMID- 11392724 TI - Real story in CalPERS talks lies beyond the headlines. PMID- 11392725 TI - Demand's cool for primary care physicians. PMID- 11392726 TI - Genetic medicine. Powerful opportunities for good and greed. PMID- 11392727 TI - How to challenge Medicare payments. PMID- 11392728 TI - Use of hospitalists: another case of 'may' vs. 'must'. PMID- 11392729 TI - He connects health care's dots along information superhighway. Interview by Patrick Mullen. PMID- 11392730 TI - A critical review of the arguments for insurance coverage for smoking-cessation therapies. AB - This article elucidates the reasons most insurance companies do not cover smoking cessation therapies despite their obvious benefits. It critically reviews the arguments for and against using a universal mandate as a strategy to increase use of smoking-cessation programs to realize the associated health benefits and cost savings. While convincing arguments exist to mandate insurance coverage for self destructive health behaviors, their merit is tempered by several valid counter arguments. For example, insurance coverage for small, routine, and predictable events, such as smoking-cessation treatment, violates the "first principles" of what ought to be covered when considered from the traditional insurance perspective. An insurance solution to risky behaviors may be to make undesirable behaviors more undesirable to individuals by raising premiums rather than to make them less undesirable with subsidies. PMID- 11392731 TI - Medicine's no place for 'artistic' sensibility. PMID- 11392732 TI - Number of uninsured unchanged; policy proposals aim to reduce ranks. PMID- 11392733 TI - The ventilator-assisted children home program. Supporting families. PMID- 11392734 TI - Pediatric home care. A cost benefit and cost effectiveness update. PMID- 11392735 TI - Staffing model for home care. PMID- 11392736 TI - Supreme Court decision paves way for home care. PMID- 11392737 TI - First star. A new approach to the fight against child abuse & neglect. PMID- 11392738 TI - HCFA responds to HAA Medicare hospice benefit questions. PMID- 11392739 TI - Search and seizure: a nonprofit health care agency stands up to government agents. PMID- 11392740 TI - Pediatric home care: nursing the shortage. PMID- 11392741 TI - Welcome to our home. An open letter to home care professionals. PMID- 11392742 TI - Assessment of heavy metal cations in sediments of Shing Mun River, Hong Kong. AB - The extent of heavy metal cation contamination in the Shing Mun River has been assessed. Sediment samples were taken at eight strategic locations along the river system. The highest concentrations of copper (Cu, 1.66 mg/g), lead (Pb, 0.354 mg/g), zinc (Zn, 2.2 mg/g) and chromium (Cr, 0.047 mg/g) were found in the Fo Tan Nullah, a major tributary of the Shing Mun River. The highest concentrations of aluminum (114 mg/g) and cadmium (Cd, 0.047 mg/g) were found in the Shing Mun Main River Channel. These contaminated sediments, accumulated over the years on the river bed, could act as secondary sources of pollution to the overlying water column in the river. PMID- 11392743 TI - Chemical contamination of the East River (Dongjiang) and its implication on sustainable development in the Pearl River Delta. AB - A recent survey reveals that water quality and river sediments of Dongjiang (the East River) have been mildly contaminated by heavy metals and organic chemicals, such as copper, lead, PCBs, PAHs and HCHs. According to photogrammertic surveys, the sources of contamination are closely related to human activities, such as domestic and industrial discharges, agricultural chemical applications and soil erosion due to deforestation. The level of water and sediment contamination is most serious at the mid-river section near the urban of Weizhou. Due to natural dilution and diffusion, the levels of contamination are slightly decreased at the medium low section of the River near Qiaotou and Matan. Nevertheless, the levels of contamination increase again in the lower river section near Shenzhen, which is a highly developed economic zone. Since Dongjiang is the major source of potable water supply for Hong Kong (nearly 80% of potable water of Hong Kong is abstracted from Dongjiang) and the other parts of the Pearl River Delta (PRD), such organic and inorganic pollution merit concern. Ironically, most of the industrial and commercial activities in the watershed of Dongjiang are business investments of Hong Kong citizens. Cross-border environmental efforts should be enhanced with collaboration of different jurisdictions to achieve targets of regional sustainability. PMID- 11392744 TI - Comparative studies on the concentration of rare earth elements and heavy metals in the atmospheric particulate matter in Beijing, China, and in Delft, The Netherlands. AB - Atmospheric particulate matter (APM) was collected at three sampling sites in Beijing, China, from February to June 1998. The concentrations of rare earth elements (REE) and cobalt (Co), zinc (Zn), copper (Cu), cadmium (Cd) and lead (Pb) in the APM were determined by inductively coupled plasma mass spectrometry (ICP-MS). The results obtained in Beijing, China, were compared to that obtained in Delft, the Netherlands, in 1997. The influence of coal combustion was considered. The results demonstrated that the content of APM, the concentrations of REE and Co, Zn, Cd, Pb in the APM in Beijing, China, were higher than that in Delft, the Netherlands. From the ratios of La to Ce, and La to Sm, which may be used as tracers for the origin of the REE, it is concluded that the origins of REE in China differ from those in the Netherlands. PMID- 11392745 TI - Microbial ecology of soils surrounding nuclear and thermal power plants in Taiwan. AB - This paper reports a study of the effect of three nuclear and one thermal power plants on the microbial ecology of soils. Populations of bacteria, actinomycetes, fungi, cellulolytic microbes, phosphate-solubilizing microbes and nitrogen-fixing microbes in the soil in the vicinity of each plant were studied. Soils were acidic at three sites, and moisture contents of the power plant soils were lower than those of the surrounding areas. Microbial populations of the topsoils (0-20 cm deep) were higher than the subsoils (21-40 cm deep), and only 10-15% of them showed significant difference (P < .05). Thirty-three percent of the samples from the surrounding areas had higher microbial population than those from the power plant areas, but 19% was the reverse. Populations of cellulolytic, phosphate solubilizing and nitrogen-fixing microbes varied with sampling locations, season and environmental conditions. Ratios of cellulolytic, phosphate-solubilizing and nitrogen-fixing microbes to total viable counts in some samples of the surrounding areas were significantly higher than in the power plant areas. Although the microbial populations of power plant soil and its surrounding area were somewhat different, it cannot be attributed as an effect of power plant operation, as the differences were not consistent. PMID- 11392746 TI - Persistence of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/F) in Ya Er Lake area, China. AB - The concentrations of polychlorinaxed dibenzo-p-dioxins and dibenzofurans (PCDD/F) in surface sediment, soil, human hair, and fish muscle from Ya-Er Lake area, China, were analyzed. The results showed that there were very high concentrations of PCDD/F existing in these samples. The results also indicated that Ya-Er Lake, which received a large amount of waste water from a nearby chloroalkali plant, was heavily polluted by PCDD/F. The present study demonstrated that those congeners, which possess at least three chlorine atoms in the lateral position with a fourth chlorine atom in the neighborhood bond of the third single chlorine atom, such as 1,2,7,8-TCDF and 2,3,6,7-TCDF, were very resistant to biodegradation due to the "neighbor effect" of every two chlorine atoms. The present study suggested that human hair may be a suitable alternative bioindicator for detecting PCDD/F exposure. PMID- 11392747 TI - A practical application of Driscoll's equation for predicting the acid neutralizing capacity in acidic natural waters equilibria with the mineral phase gibbsite. AB - A practical application of Driscoll's equation for predicting the acid neutralizing capacity (ANC) in acidic waters equilibria with the mineral phase gibbsite is reported in this paper. Theoretical predication values of ANC are compared with the experimental data obtained from different literatures. The effect of aluminum (Al) on the value of ANC is investigated. It indicates that Al plays an important role in regulating the buffering effects in acidic natural waters. Failure to consider Al in acidic waters may bias assessment results in certain situations so as to overestimate the ANC values in response to increase in atmospheric deposition. PMID- 11392748 TI - Selenium geochemistry of paddy soils in Yangtze River Delta. AB - Selenium (Se) content both in total and bioavailable forms were very low (25-35 and 9.5 micrograms/kg, respectively) in waterlogged paddy soils in the central Yangtze River Delta. This was due to changes in geochemistry and the inherent properties of the parent materials of these lowland paddy soils. The very low Se content (23 micrograms/kg) in the parent material resulted in low total Se content in the soil. The main chemical changes under long-term waterlogged conditions are depletion of molecular oxygen, decrease of redox potential, and reduction of Fe (III) to Fe (II) and SeO3(2-) to Se0. This led to low availability of Se in soils, and subsequently low Se content (29 micrograms/kg) in brown rice grain produced in this region. It has been suspected that low Se content in staple food might be one of the major reasons for a high infection rate of the intestine and stomach cancers and the higher death rate caused by these diseases in the region. Foliage spray of Na2SeO3 at early heading stage of rice plant growth improved the Se content of brown rice grain, hull, and straw, and would improve human and animal health. PMID- 11392749 TI - Distribution characteristics of fluoride and aluminum in soil profiles of an abandoned tea plantation and their uptake by six woody species. AB - Distribution characteristics of fluoride (F) and aluminum (Al) in soil profiles of an abandoned tea plantation in Hong Kong and their uptake by six woody species namely Camellia sinensis, Melastoma affine, Sterculia lanceolata, Ardisia crenata, Acacia formosa and Machilus thunbergii were investigated. C. sinensis accumulated both F (656 mg/kg in the leaves) and Al (8910 mg/kg in the leaves). M. Affine only accumulated Al (9932 mg/kg in the leaves), while S. lanceolata, A. crenata, A. formosa and M. Thunbergii were Al (26-115 mg/kg in the leaves) and F (20-25 mg/kg in the leaves) excluders. Water-soluble F and Al contents accumulated in surface soils (especially 0-3 cm) and decreased with depth along soil profiles: A layers (2.0 +/- 0.6 and 31 +/- 5.8 mg/kg) > B layers (1.5 +/- 0.2 and 27 +/- 3.5 mg/kg) > C layers (1.3 +/- 0.2 and 20 +/- 2.7 mg/kg), respectively. The water-soluble, NH4Ac-extractable and HCl-extractable F in soil profiles were significantly correlated (P < .01). F concentrations in all soil layers (n = 51) of soil profiles (n = 17) were significantly correlated (P < .01) to Al concentrations in terms of water-soluble Al/F, HCl-extractable Al/F and NH4Ac-extractable Al/F, due to the formation of Al-F complexes in soil. F contents extracted with deionized water (pH = 6.68), 1 mol/l HCl and 1 mol/l NH4Ac in soil could be used to predict F concentrations in the leaves, stems and roots of the six plants. PMID- 11392750 TI - Location and migration of cations in Cu(2+)-adsorbed montmorillonite. AB - Locations of Cu2+ ion in Cu(2+)-adsorbed montmorillonite have been studied by electron paramagnetic resonance (EPR), supplemented by X-ray diffraction (XRD) and differential thermal analysis (DTA). In the EPR spectra of Cu(2+)-adsorbed montmorillonite, three signals, corresponding to Cu2+ ion, have been simultaneously recorded. Some Cu2+ ions seemed to replace the original interlayer metal cations and some entered into the hexagonal cavities. A small fraction of Cu2+ ions penetrated into the octahedral vacancies. There were two ways for the adsorption of Cu2+ ion by montmorillonite--exchangeable and specific. On heating, the hydrated Cu2+ ion in the interlayer loses the coordinating water and then enters into the hexagonal cavities. When the heating temperature further increased, dehydroxylation occurs, which facilitates Cu2+ ion in the hexagonal cavities to penetrate into the octahedral vacancies. PMID- 11392751 TI - Effect of several organic acids on phosphate adsorption by variable charge soils of central China. AB - The effect of several organic acids on phosphate adsorption by acidic soils in subtropical zone of central China was studied. Results showed: (1) citrate and oxalate remarkably reduced the amount of phosphate adsorption, but tartrate, benzoate and acetate had only a very slight influence on phosphate adsorption; (2) the ability of citrate in reducing phosphate adsorption was greater than that of oxalate, moreover, the reduction percentage was dependent on the concentration of organic ligands in the solution; (3) the effect of organic acids on phosphate adsorption was related to the pH value of organic acid solution. The minimum reduction in adsorption of phosphate was present at a specific pH value of organic acid solution which ranged from 2 to 10; (4) a minimum reduction of phosphate adsorption occurred when phosphate was added to the soils before organic acid, whereas a maximum occurred when organic acid was introduced before the addition of phosphate. Meanwhile, the treatment for the mixture of two organic acids resulted in more reduction in phosphate adsorption than each of the organic acids and less than the total of them. Based on these observations, we suggested that the competition between phosphate and organic acids relied on their relatively affinity to soil mineral surface at different conditions. PMID- 11392752 TI - Behaviour of chemical elements during weathering of pyroclastic rocks, Hong Kong. AB - The behaviour of whole-rock major, trace and rare earth elements (REE) during weathering under subtropical conditions is examined along a profile developed over crystal--vitric tuffs with eutaxitic texture. The intensity of weathering within the profile varies erratically, indicating weathering processes operate over different scales. Quartz, K-feldspar, plagioclase and biotite are the main primary minerals, whereas clays, sesquioxides, sericite and chlorite are the alteration products. Kaolinite, halloysite and illite-mica are the dominant clay minerals present in significantly varying proportions. Two competing processes, namely leaching and fixation, are the main regulators of variations in mostly major and some trace element concentrations along the profile. In general, as the intensity of weathering increases, Ca, Na, K, Sr +/- Si decrease, while Fe, Ti, Al and loss of ignition (LOI) increase. Likewise, the intensity of negative Eu anomaly decreases while the intensity of negative Ce-anomaly and the La/Lu and Sm/Nd ratios increases. In detail, however, the behaviour of chemical elements cannot be solely explained in terms of the degree of weathering. This study makes it clearly evident that the type and abundance of sesquioxides and clay minerals can significantly modify the geochemical signatures of weathering processes. PMID- 11392753 TI - Nutrient release and sediment oxygen demand in a eutrophic land-locked embayment in Hong Kong. AB - Tolo Harbour is a large eutrophic land-locked estuarine embayment in Hong Kong. The rapid urbanization, commercio-industrial activities and lack of legislative control around the Tolo catchment produced large quantities of untreated or partially treated municipal sewage, agricultural wastes and cottage industrial effluents which were discharged into Tolo Harbour via rivers and watercourses. Control measures were implemented to reduce the external nutrient loading into the harbour since the early 1980s. Nutrient data for the period 1982 to 1997 were analyzed for temporal trends. Over the period of observation, the total inorganic nitrogen and total phosphorus both show an increasing trend, despite a decade of efforts in reducing nutrient loading. The release rates of potentially mobile nitrogen (N) and phosphorus (P) from the sediments collected from Tolo Harbour were determined by N and P release experiments under oxic conditions. The experimental results showed that the sediment released significant amount of nutrients, especially orthophosphates and ammonia nitrogen. The maximum release rates were 15.0 and 206.0 mg/m2/day, respectively. Although the external nutrient loading has been reduced, nutrients could gradually be released back into the water column from the contaminated sediments and delay improvement of the water quality. PMID- 11392754 TI - Dynamics of aluminum speciation in forest-well drainage waters from the Rhode River watershed, Maryland. AB - This paper reports an investigation of the dynamics of aluminum (Al) speciation in the forest-well waters from study site 110 of the Rhode River watershed, a representative sub-unit of Chesapeake Bay. Seasonal changes of Al speciation are evaluated by a modified MINEQL computer model using chemical equilibrium calculation. It was found that Al-F and Al-Org complexes were the dominate forms, whereas toxic forms of Al3+ and Al-OH were not significant. This indicates that Al toxicity is not very serious in the Rhode River area due to the high concentrations of fluoride and organic materials, even though sometimes pH is very low (approximately 4). Increased H+ or some other associated factors may be responsible for the decline in fish and amphibian population on the watershed. PMID- 11392755 TI - A flow system for the determination of metal speciation in soil by sequential extraction. AB - A flow extraction system with on-line and off-line flame atomic absorption spectrometric (FAAS) detection was developed to speed up, facilitate, and improve the accuracy of sequential extraction for metal speciation in solid materials. A flow extraction system with off-line detection was more advantageous than the on line detection. In the proposed system, extraction was performed in a closed extraction chamber where extractants were flowing through sequentially. The extract from each extraction was collected in 4-10 fractions to obtain a total of approximately 180 ml for subsequent FAAS determination. The system is simple, easy to construct and operate. It has less risk of contamination and human error. A widely used three-step sequential extraction scheme was used to evaluate the novel system by analyzing Ca, Fe, Mn, Cu, and Zn in a certified reference material. The extraction time for three-step sequential extraction of soil sample was 4 h. The analytical results for Ca, Fe, Mn, Cu, and Zn of a soil certified reference material using the proposed system were compared with those of the conventional batch extraction. PMID- 11392756 TI - Acidification of lead/zinc mine tailings and its effect on heavy metal mobility. AB - The acid-forming potential of lead/zinc (Pb/Zn) mine tailings at Lechang City of Guangdong Province was studied using both net acid generation (NAG) and acid-base accounting (ABA) methods. The pyritic and total sulfur contents of the tailings were 12.6% and 18.7%, respectively. The mean acid neutralization capacity (ANC) was 63.5 kg H2SO4/t while three oxidized tailings samples had an ANC less than zero. The NAG and net acid production potential (NAPP) values were 220 and 326 kg H2SO4/t, and both the NAG and NAPP results indicated that the tailings had a high acid-forming potential. NAG was more accurate than NAPP in predicting acid forming potential of the tailings due to uncompleted oxidization of pyritic sulfur. Analysis of samples from two profile tests indicated that acidification mainly occurred at the surface (0-20 cm) and had little effects at deep layer of the tailings. Total concentrations of Pb, Zn, Cu, and Cd were increased greatly with depth at the acidified tailings profile, while heavy metal concentrations at different depths of nonacidified tailings profile were similar. The results indicated that depletion of heavy metals at the acidified surface was due to acidification. The diethylenetetramine pentaacetic acid (DTPA)-extractable Pb, Zn, Cu, and Cd concentrations of acidified tailings surface (0-20 cm) were significantly higher than those of nonacidified tailings, which further revealed that acidification enhanced the mobility of heavy metals in the tailings. PMID- 11392757 TI - Removal of copper from water using limestone filtration technique. Determination of mechanism of removal. AB - This paper discusses heavy metal removal from wastewater by batch study and filtration technique through low-cost coarse media. Batch study has indicated that more than 90% copper (Cu) with concentration up to 50 mg/l could be removed from the solution with limestone quantity above 20 ml (equivalent to 56 g), which indicates the importance of limestone media in the removal process. This indicates that the removal of Cu is influenced by the media and not solely by the pH. Batch experiments using limestone and activated carbon indicate that both limestone and activated carbon had similar metal-removal efficiency (about 95%). Results of the laboratory-scale filtration technique using limestone particles indicated that above 90% removal of Cu was achieved at retention time of 2.31 h, surface-loading rate of 4.07 m3/m2 per day and Cu loading of 0.02 kg/m3 per day. Analyses of the limestone media after filtration indicated that adsorption and absorption processes were among the mechanisms involved in the removal processes. This study indicated that limestone can be used as an alternative to replace activated carbon. PMID- 11392758 TI - Adsorption of phenol on inorganic-organic pillared montmorillonite in polluted water. AB - Both inorganic- and organic-pillared montmorillonites (PMts) were used to adsorb phenol to study suitable conditions for adsorption and adsorption isotherms. The adsorbing capacity of modified clays depends not only surface area, but mainly on micropore structure and surface components. After incandescing at 500 degrees C, the pillar structure and the basal interlayer spacing (1.83 nm) remained stable. Using modified PMt with surfactant can improve adsorbing capacity greatly. The PMt can be recycled, and it is a potential substance for adsorption of environmental pollutants. PMID- 11392759 TI - Bioremediation of oil sludge-contaminated soil. AB - Bioremediation has become an important method for the restoration of oil-polluted environments by the use of indigenous or selected microbial flora. Several factors such as aeration, use of inorganic nutrients or fertilizers and the type of microbial species play a major role in the remediation of oil-contaminated sites. Experiments were undertaken for bioremediation of oil sludge-contaminated soil in the presence of a bacterial consortium, inorganic nutrients, compost and a bulking agent (wheat bran). Experiments were conducted in glass troughs for the 90-day period. Bulked soil showed more rapid degradation of oil compared to all other amendments. During the experimental period, wheat bran-amended soil showed 76% hydrocarbon removal compared to 66% in the case of inorganic nutrients amended soil. A corresponding increase in the number of bacterial populations was also noticed. Addition of the bacterial consortium in different amendments significantly enhanced the removal of oil from the petroleum sludge from different treatment units. PMID- 11392760 TI - Utilization of petroleum hydrocarbons by Pseudomonas fluorescens isolated from a petroleum-contaminated soil. AB - A strain of Pseudomonas fluorescens, isolated from petroleum hydrocarbon contaminated soil was examined for its ability to utilize a variety of hydrocarbon substrates. Surface tension measurements indicated the production of biosurfactant during the microbial degradation of hydrocarbon. The organism utilized both short and long chain n-alkanes. It emulsified a number of aliphatic and aromatic hydrocarbons. PMID- 11392761 TI - Relationships between chromium biomagnification ratio, accumulation factor, and mycorrhizae in plants growing on tannery effluent-polluted soil. AB - Heavy metal-contaminated land is increasingly becoming an important environmental, health, economic, and planning issue in Pakistan. The unplanned disposal of industrial effluent from tannery, for example, has resulted in a many fold increase in chromium (Cr) in the land near a tannery. This study was undertaken to compare the total and the DTPA-available Cr contents in the soil and the roots and leaves of tree species growing on it with those on the nearby noncontaminated reference site at Kala Shah Kakoo, Panjab, Pakistan. A very reduced plant cover on the tannery effluent-contaminated site was noted and there was a sharp boundary between the polluted and nonpolluted reference sites, suggesting a strong selection pressure. Polluted soil contained considerable higher amounts of Cr as compared to the reference soil but no correlation was found between Cr contents in the dried plant tissue and the total DTPA extractable Cr. Roots of all the three tree species, i.e. Dalbergia sissoo, Acacia arabica, and Populus euroamericana, growing on both the contaminated as well reference site possessed arbuscular mycorrhizal fungal (AMF) infection in their roots and AMF propagules in the associated rhizospheres. D. sissoo and A. arabica roots were also studded with nitrogen-fixing rhizobial root nodules, while those of P. euroamericana possessed AMF as well as ectomycorrhizal infections. The dual infection would encourage mineral nutrition, including Cr. AMF community varied, i.e. trees growing on the reference site were exposed to a wide variety of AMF such as Glomus, Scutellospora, and Acaulospora, whereas those on the contaminated site contained only Gigaspora spp. in their mycorrhizospheres, suggesting a selection pressure. Typical Glomus infection patterns in the roots of D. sissoo growing on the contaminated soil but absence of spores of Glomus spp. in the associated rhizospheres indicate the potential error of using AMF spores to extrapolate the root infection. High Cr contents adversely affected the size, diversity, and species richness of AMF as measured by Shannon-Weiner index. The potential of mycorrhizae in protecting the host plant against the harmful effect of heavy metals and in phytoremediation of the Cr-polluted soil is discussed. PMID- 11392762 TI - Oxygen demand, nitrogen and copper removal by free-water-surface and subsurface flow constructed wetlands under tropical conditions. AB - This study was conducted to: (1) assess the role of wetland vegetation in the removal of oxygen demand and nitrogen under tropical conditions, (2) estimate the uptake of nitrogen and copper by wetland plants and (3) investigate the speciation of Cu in wetland media among four operationally defined host fractions, namely exchangeable, carbonate, reducible and organically bound. Four laboratory-scale wetland units, two free-water-surface (FWS) and two subsurface flow (SF) with one of each planted with cattails (Typha augustifolia), were fed with primary-treated sewage and operated at nominal retention times of 0.6-7 days. The influent and effluent BOD/COD and nitrogen concentrations were monitored to assess the performance of the wetland units for various mass loading rates. At the end of the study, all cattail plants were harvested and analyzed for total Kjeldahl nitrogen (TKN). Four other wetland units, which were identical to the first four, were fed with domestic wastewater spiked with copper in increasing concentrations. Copper speciation patterns in the sand layer were determined at the end of the study. The results showed that wetland vegetation did not play an important role in oxygen demand removal but were capable of removing about 22% and 26% of the nitrogen input in the FWS and SF wetland units, respectively. Mass balance analysis indicated that less than 1% of copper introduced was taken up by the cattails. Copper speciation patterns in the sand media showed that the exchangeable fraction contributed 30-57% and 63-80% of the nonresidual copper in the planted and unplanted FWS wetlands, respectively. For SF units, the percentages were 52-62% and 59-67%, respectively. This indicates that large amount of copper in the media were potentially remobilizable. PMID- 11392763 TI - An assessment of the impact of motorway runoff on a pond, wetland and stream. AB - The impact of soil filtered runoff from a section of the M25 outer London motorway (constructed in 1981) on a pond, wetland and stream in a nature reserve was investigated by monitoring water, sediment. The tissues of the emergent plants Typha latifolia and Glyceria maxima collected from the pond were analysed for the heavy metals, Cd, Pb, Cu and Zn. Macroinvertebrates were monitored in the stream and biotic indices applied to the data. The plant tissue concentrations for Typha and Glyceria show decreasing metal concentrations from root to rhizome to leaf. This trend has previously been reported for Typha exposed to runoff although the tissue concentrations are lower in this study with the exception of Cd in root tissue. The Biological Monitoring Working Party (BMWP) score and Average Score Per Taxon (ASPT) for the stream at sites above and below the pond outlet are lower than the scores recorded by the Environment Agency for England and Wales at an upstream site above the Pond/Wetland. The sites have an Overall Quality Index of 'moderate water quality', and there is no evidence of a deterioration of biologically assessed water quality between them. The results of the study show the long-term impact on sediment of filtered road runoff discharges to a natural wetland and pond located in a nature reserve. The use of natural wetlands for the discharge of road runoff is inadvisable. Constructed wetlands in combination with other structures including settlement trenches and ponds should be considered as an alternative treatment option. PMID- 11392764 TI - Artificial wetlands and water quality improvement. AB - This paper illustrates the role of plants to assist the treatment of water pollution in man-made wetlands in tropical and temperate climates. It also considers the potential for environmental education of these wetland systems. The management and natural treatment of pollution is described in the Mai Po Marshes, Hong Kong and a wetland in London which is also an important site for birds. The design of the Putrajaya Lake and Wetland system in Malaysia is compared with a constructed wetland and lake for the treatment of urban surface runoff in a new residential development in the United Kingdom. The benefits of these natural systems are discussed in the context of the global trend for introducing sustainable methods of environmental management and low cost pollution treatment systems. PMID- 11392765 TI - Growth response of Sesbania rostrata and S. cannabina to sludge-amended lead/zinc mine tailings. A greenhouse study. AB - Legumes are ideal for revegetation of metal-mined wastelands which lack nitrogen (N). A greenhouse study was conducted to investigate the feasibility of using Sesbania rostrata and S. cannabina for the reclamation of lead/zinc (Pb/Zn) mine tailings and to evaluate the effects of organic amendment using sewage sludge (0%, 25%, 50%, and 75%, v/v). The results showed that both species could continue to grow on the highly toxic tailings substrata for at least 80 days, although their growth suffered from adverse effects. That S. rostrata with stem and root nodules had better growth (biomass, growth rates, and biomass of nodules) than S. cannabina suggested that S. rostrata is a better choice as a pioneer species for revegetation of the mine tailings. Stem nodules had less obvious adverse effects imposed by tailings than root nodules. Application of sewage sludge increased contents of total carbon (C), N, phosphorus (P), and potassium (K), and reduced total Zn, Pb, Cd, and DTPA-extractable Pb and Cd in tailings substrata. These, in turn, reduced metal (Zn, Pb, and Cd) uptake and accumulation in plant tissues, and improved plant growth performance, including biomass, growth rates, stem nodulation. Fifty percent (v/v) of sludge application rate was the best loading rate for plant growth. PMID- 11392766 TI - [Optimization of the process of culturing yeast in the presence of the dispersed mineral palygorskite]. AB - A positive effect from introduction of the highly dispersed mineral palygorskite in the nutrient medium for aerobic batch cultivation of the yeast Saccharomyces cerevisiae, the Kyivsky strain, used in production of champagne has been established experimentally. An adequate mathematical model describing the process of growth has been constructed. Optimal cultivation conditions have been determined which provided for the achievement of such yeast cell concentration in the culture fluid as 116 mln/ml, palygorskite concentration in the nutrient medium being 0.37 g/l and sucrose 2.0 +/- 0.2%. The yeast obtained possess higher fermentation activity. PMID- 11392767 TI - [Genetic transformation of Streptomyces globisporus strain 1912: restriction barrier and plasmid compatibility]. AB - Low efficiency of genetic transformation of protoplasts of different strains of Streptomyces globisporus 1912 by means of DNA preparations of three vectors pIJ487, pGM160 and pWHM4 is explained by presence of the restriction barrier in the recipients. This obstacle can be overcome by the use of modified DNA of the same vectors, isolated from not numerous transformants. The frequency of transformation by such modified vector DNA was increased by two-three orders in comparison with initial DNA, isolated from Streptomyces lividans TK24, loosing restriction-modification system. The vector pCNB4001, containing the replicon of endogeneous plasmid pSG1912-1, effectively transformed the protoplasts of all S. globisporus 1912 strains. Compatibility of pIJ487 and pSG1912-1 plasmids and incompatibility of the latter and pWHM4 was shown. PMID- 11392768 TI - [Adsorbed receptors for Erwinia carotovor subsp. carotovora macromolecular bacteriocins]. AB - Study of nature of receptors for macromolecular bacteriocins Erwinia carotovora subsp. carotovora has shown that lipopolysaccharide (LPS) of cell membrane is an attaching structure for them. It has been established that enzymic treatment of LPS preparation with its further deproteinization by phenol is necessary for isolation of biologically active lipopolysaccharide. The process of absorption by LPS has been studied quantitatively and it has been shown that it is a low efficient receptor as compared with LPS included in the native cell membranes. An approach has been proposed for the first time to the estimation of monosaccharide composition of LPS-receptor based on relations between bacteriocin sensitivity and content of monosaccharides. Study of six strains of E. carotovora subsp. carotovora from different sources has shown that the structure of LPS-receptors includes mannose, fucose, xylose, ramnose and two lipophylic monosaccharides of unknown nature. A conclusion has been made that S-LPS (0-chain) is that part which contains the sites of attachment of macromolecular carotovoricins. PMID- 11392769 TI - [Amino acid and lipid composition of streptomycetes biomass, isolated from Moldovian soil]. AB - The aminoacid and lipid composition of the biomass of streptomycetes isolated from soils of Moldova was studied. The strains tested being cultivated on complex medium, the biomass of streptomycetes contained 41-64.2% of irreplaceable aminoacids from the total amount of the found aminoacids. The amount of phospholipid and sterene fractions in the lipids of some strains was twice as high as in the already studied collection cultures. A wide spectrum of fatty acids (from C15:0 to C22:n) with prevalence of unsaturated ones was also observed. PMID- 11392770 TI - [Serologic characteristics of Pseudomonas fluorescens strain 7769, isolated from rye]. AB - Serological properties of Pseudomonas fluorescens 7769, isolated from the damaged tissues of rye have been studied. It has been shown that P. fluorescens 7769 was serologically related but not identical as to antigenic composition with the type strain P. fluorescens 4125 of biovar 1, to which it belonged and closely related to the strain P. fluorescens 1371, isolated from wheat. Availability of weakly expressed common antigens of P. fluorescens 7769 with the agent of the disease of grain cultures P. syringae pv. atrofaciens has been established. PMID- 11392771 TI - [Invasion of microorganisms in bronchial mucosa of liquidators of the Chernobyl accident consequences]. AB - Bronchial bioptates of 97 liquidators of the Chernobyl accident consequences with chronic bronchitis and 23 patients of control nosological group as well as sputum (174 persons) and BAL (22 persons) of liquidators with chronic obstructive lung disease (COLD) were studied to define pathogenic role of automicroflora in the development of lung diseases. Such methods as electron microscopy, immunohistochemistry and microbiology were used. The revealed invasion of microorganisms occurred against the background of pathology of superficial bronchial epithelium with a decrease of HLA-DR and CD23 lymphocytes and increase of CD1c lymphocytes in lamina propria of bronchial mucosa of the liquidators. Verification of microorganisms characteristic of the upper respiratory tracts and atypical presence of Escherichia coli were found in the contents of the lower parts of broncho-pulmonary system of the liquidators. The obtained results testify to the activation of automicroflora and appearance of pathogenic microorganisms were caused by deterioration of specific and non-specific immune protection in liquidators with COLD. PMID- 11392772 TI - [Biotechnology for purifying surface storm waters]. AB - A possibility of purification of the surface storm drains from petroleum has been shown. The developed extensive biotechnology is based on the use of the preparation "Desna"--an active destructor of hydrocarbons. The application of the biotechnology at the plants for treatment of the surface storm waters from the industrial zone Telychka of the city of Kyiv has allowed the content of petroleum in water dropped to the Dnieper to be constantly reduced 50-100 times. PMID- 11392773 TI - [Deposition of microorganisms at patenting the invention and results for the inventor]. AB - The standards of the patent legislation of Ukraine and other countries concerning the deposition of microorganisms at patenting of the invention and conditions for the availability of the deposited samples have been considered. The questions connected to practice of strains deposition in Ukraine which require the normative decision or specification in view of the tendency of development of patent legislation in the countries advanced in the sphere of the industrial property have been outlined. PMID- 11392774 TI - [Conceptual basis for defining viral insecticide harmlessness for human health and the environment]. AB - The qualitative distinction of the new class of virus-based biopesticides from chemicals is the intracellular parasitism of the active agent in the organisms of the living biological host, its capacity to join natural microbiocenoses and to restrain the number of the destructive species over a long period of time. Thus, the program of the study of harmlessness of entomopathogenic viruses must be aimed at preventing the possibility of their adaptation to non-target animals, reproduction and the ability to provoke the non-specific infection. Other parameters such as toxicity and allergenicity of the preparations and their standardization will be investigated under the absence of such ability. The appropriate schemes and methods of investigations have been developed. PMID- 11392775 TI - [Bacteria of Bacillus species--prospective source for biologically active substances]. AB - The present data from literature on bio-active substances produced by aerobic sporulating bacteria of Bacillus genus are presented. Problems of antibiotics synthesis by bacilli are considered. The antibiotics under consideration are both well-known and new ones--biosurfactants characterized by the surface-active properties. Enzymic activity of Bacillus genus bacteria has been characterized. Special attention was given to the analysis of enzymes, which could play a positive part in the digestion processes. Data are presented which evidence that the bacilli produce the substances, which affect essentially the immune reactivity of the macroorganism. PMID- 11392776 TI - [Metabolic disturbances and correction of them in alcoholic and substance abusing people]. PMID- 11392777 TI - [Research of the academician I. N. Bulankin on problems of protein chemistry]. AB - The article presents an information about scientific researches on the problem of chemical properties and structure of proteins, which were carried out intensively by the biochemists of Kharkiv State University in 30-50 years of XX cent., headed by I. N. Bulankin, academician of Ukrainian Academy of Sciences. This researches dealt with the processes of proteolysis and protein denaturation, the peculiarities of globular and fibrilliar proteins behavior in solution, the properties of complexes with proteins and nucleic acids, etc. The historic analysis of origin and development of this scientific problem at the biochemistry chair of the university and its gradual replacement by researches with other biochemical problems has been carried out. The data concerned with the general situation in the field protein structure of estimation at the world and home biochemistry are cited too. In connection with the above presented the authors give the grounded conclusions about the importance of the scientific researches carried out by I. N. Bulankin as well as about. PMID- 11392778 TI - [Polypeptide serine proteinase inhibitors isolated from venom from several reptilian species]. AB - The survey encompasses literature data on the polypeptide inhibitors of some reptiles serine proteinases and their separation from adder Viperidae and cobra Elapidae species. The evolutionary comparison of physico-chemical and biological properties of them are also given and discussed within this work. Considerable homology (about 50%) in amino acid composition of adder, bee, mammal and others of different phylogenetic origin is being emphasized and high homology in structure of their functionally important inhibitors sites is observed. In the most cases the investigated peptide inhibitors of adder and cobra were observed to have an extremely high antitryptic activity with Ki ranging from 7.6 x 10(-10) M to 3.5 x 10(-12) M. The majority of polypeptide inhibitors are suggested by Laskowsky et al to interact with the proteinases in a standard way. The biological reactivity of the above preparations is a result of arginine and lysine presence in the substrate-binding sites of P2' and P3' or P4' centres. PMID- 11392779 TI - [Role of formic acid in transformation of acetic and succinic acids in animal tissues]. AB - Acetic and succinate acids KoA acyl derivatives interacting with formate were displayed to produce alpha-ketoacids--pyruvate and alpha-ketoglutarate. These acids also interact with formate and make pyruvic and malate acids, while alpha ketoglutarate, evidently, tricarboxy acids. Interaction of formate with acetic and succinate acids inspite of occurring out of the tricarbone cycle increases the latter metabolic functions. PMID- 11392780 TI - [Effect of primary aliphatic amines on oxidative phosphorylation in rat liver mitochondria]. AB - The dynamics of primary aliphatic amines (ethylamine, propylamine) effects on the processes of oxidative phosphorylation in rat liver mitochondria was estimated. The inhibiting action of ethylamine and propylamine on the oxidative phosphorylation processes in the rat liver mitochondria was revealed. PMID- 11392781 TI - [Effect of calmodulin on myometrial cell membrane Ca2+,Mg2+-ATPase activity]. AB - Myometrium cell plasma membrane Ca2+, Mg(2+)-ATPase purified by an affinity chromatography on calmodulin-sepharose 4B is calmodulin-dependent enzyme. Concentration of calmodulin required for half-maximal activation of enzyme was about 26 nM. By unlike to the enzymes originated from other tissues sensitivity to the calmodulin of the myometrial sarcolemma Ca(2+)-transporting ATPase was lower: calmodulin increased Vmax of ATPase about 1.25-fold, the apparent constant of the activation of enzyme by Ca2+ failed to alter independently on the phospholipid presenting at the enzyme isolation. PMID- 11392782 TI - [Electronic structure and energetics of transitions in absorption spectra of hypericin]. AB - In half-empirical sigma, pi-valent approximation (PM3) the energetics of tautomer transitions of hypericin and nature of electronic transitions in experimental and calculation absorption spectra is investigated. On the basis of quantum-chemical researches results concerning five tautomer condition of hypericin is established that the most stable is its known structure of a quinone I (7,14-dion 1,3,4,6,8,13-hexahydroxydianthrone). Is determined that longwave characteristic bands in hypericin experimental absorption electronic spectra are connected with its electron- and proton-donor, and acceptor properties, and are caused by modifications of its quinone structure by formation of alcohols, cation- and anion-radicals. PMID- 11392783 TI - [Concentration of transferrin iron and transferrin saturation in blood]. AB - Transferrin iron, transferrin protein concentrations, and transferrin saturation have been determined for the first time in the whole blood. Microsamples were taken from healthy adults and patients with occupational secondary haemochromatosis using quantitative electron spin resonance technique. At elevated transferrin saturation, transferrin saturation values determined in the plasma and serum samples were shown to be less than respective values determined in the whole blood of the same patients. At increased transferrin iron concentration the difference between experimental and reference data sets determined in the blood and plasma was statistically significant in contrast to data sets determined in serum. Therefore, the analysis of the blood microsamples ensured an adequate estimation of transferrin iron concentration, especially at high transferrin saturation. A new index--transferrin iron concentration in the formed blood elements--was introduced. The values of the index were determined in the groups of healthy adults, patients with secondary occupational hemochromatosis and healthy newborns. PMID- 11392784 TI - [Structure and function of high molecular weight proteinases in human and animal cells]. AB - The new data that concern structure and functions of high-molecular-mass proteases in animal and human cells are presented in the review. The participation of ATP in mechanisms of proteolysis is considered. Considerable attention is paid to the role of high-molecular-mass multicatalytic proteinase in the regulation of intracellular metabolic processes, cell proliferation, transformation and apoptosis, its involvement in haemopoiesis and immune responses. PMID- 11392785 TI - [Characteristics of bacterial lipopolysaccharides depending on extraction method]. AB - Carbohydrate-containing biopolymers have been isolated from Erwinia carotovora subsp. atroseptica 549 by two methods--the aqueous-phenol and with using physiological solution--with addition and without addition of ethylenediaminetetraacetate (EDTA). The biopolymers yield from the cells of bacteria are shown to depend on the extraction method. Lipopolysaccharide-protein complex have been isolated by the sparing method. The purest lipopolysaccharide have been isolated by the aqueousphenol method. The preliminary treatment of cells by EDTA increased the biopolymers output. Carbohydrate containing biopolymers isolated from the cells of bacteria by different methods possess the similar qualitative composition of monosaccharides but they differ in the quantitative content of monosaccharides, spectrum of fatty acids of lipid components as well as in the protein content. PMID- 11392786 TI - Chernobyl clean-up workers erythrocyte membrane lipid content at the remote period. AB - Chernobyl Nuclear Power Plant accident clean-up workers were exposed to different radiation doses. Realization of remote effects depends on cellular membrane structure and function. We studied lipid composition of erythrocyte ghost and blood plasma in clean-up workers on the 12-th year after the accident. Groups of Chernobyl accident clean-up workers, non-irradiated patients with the same diseases and healthy controls were studied. Phospholipids were analyzed by two dimentional thin layer chromatography on silica gel. To analyse fatty acids and cholesterol gas chromatography was used. Total cholesterol and total phospholipids amounts in clean-up workers erythrocyte ghost were increased. The level of sphingomyelin and phosphatidylethanolamine raised. In some of the studied irradiated patients lysophosphatidylethanolamine appeared. The distribution of (diacyl)- and plasmalogen forms of phosphatidylcholine and phosphatidylethanolamine was not affected. The fatty acid composition changed slightly except 22:5 n-3, which level significantly increased and 22:5 n-6, which quantity lowered. The unsaturation index remains unchanged. No change in lipid composition of blood plasma was detected. All the changes in lipid composition of erythrocyte ghost of irradiated and unexposed patients were very similar. The obtained results confirm the suggestion about non-specific remote effects after exposure to low doses of ionizing irradiation. PMID- 11392787 TI - [Effect of factors from the Chernobyl exclusion zone on the level of cytochrome P 450 in mouse liver microsomes]. AB - The data concerning the influence of Chernobyl exclusive zone factors on cytochrome P-450 relative contents in liver microsome fractions of Balb/c, CC57W/Mv and C57 Black mice are submitted in this work Reliable diminishing of liver P-450 relative concentration in experimental animals, compared with respective isogenic control groups is shown to be varying depending on the mouse line and sex. PMID- 11392788 TI - [Laboratory diagnosis of the status of the hemostasis system]. AB - Activators of fibrinogen, prothrombin and protein C isolated from venoms of Agkistrodon halys halys and Echis multisquamatus may be used as a tool both for thrombosis investigations in the model systems and for diagnostic in the clinical practice. The complex of diagnostic tests developed on the base of these activators allows to characterize the haemostatic system at different pathologies and as well as to determine the unbalance between separate components of haemostasis. The tests are approved on plasma of patients with heart diseases, ulcers, nephrites, hestoses etc. The tests are sensitive, informative, easy to use and do not require an additional equipment. They have no analogues in Ukraine. PMID- 11392789 TI - [Change in indicators of peroxide resistance in erythrocytes and lipid metabolism indicators in blood plasma during development of ischemic insult]. AB - The authors have displayed that ischemic insults are accompanied by the significant intensification of free radical peroxidation processes both in erytrocyte membranes and in blood plasma. The changes mentioned are characterized by the simultaneous decrease of erytrocites resistance to peroxide hemolysis, as well as by the parallel lowering of alpha-tocopherol concentration in blood plasma. PMID- 11392790 TI - [Provision of residents of the city of Kiev with vitamins C and B1. Effectiveness of a complex vitamin-mineral preparation in preventing spring vitamin deficiencies]. AB - The investigation was conducted on provision with vitamins C and B1 of Kyiv inhabitants limited contingent at spring period and efficiency of complex vitamin mineral preparations in normalisation of the vitamins level in blood of human subjects. The deficit of vitamin C in 54% and vitamin B1 in 18.5% of the investigated people was revealed. Supradin gave the best results among rest preparations ("Centrum", "Vitrum", "Duovit", "Multi-Tabs") in the normalization of the level of vitamin C and biologically-active form of thiamine (thiamindiphosphate) in the blood of investigated people. PMID- 11392791 TI - [Proteolytic system in lungs upon inhalation exposure to low doses of lead salts]. AB - Changes in proteinase--antiproteinase activities in lungs as a result of short term (for 1 week) inhalation of rats with 0.01% (CH3COO)2 Pb have wholly compensative patterns, but it have been found increased cathepsine B activity as a negative prognostic factor. Chronic (for 2 month) toxicant inhalation caused considerable activation of both trypsine and cathepsine B under decreasing alpha 1-antitrypsine and alpha 2-macroglobuline activities. Cathepsine L activity was not affected. Disorders in the proteolysis system were evaluated as desadaptation situation in lung tissue under chronic toxic influence. PMID- 11392792 TI - [Enzymatic sensor for determing triglycerides]. AB - An enzymatic sensor based on ion selective field effect transistors was developed for the determination of triacylglyceroles in the range of concentrations from 0.1 to 2 mM. It was show that the sensitivity of the sensor was equal to 0.1 mM and time of one measurement was 2 min. The optimal environment for the triacylglyceroles registration is 1 mM sodium phosphate buffer (pH 7.3). PMID- 11392793 TI - Deprotonated carboxylic group of amino acids transforms adenine into its rare prototropic tautomers. AB - By UV spectroscopic data for anhydrous DMSO solutions and ab initio HF/6-31G** calculations in vacuum it was shown for the first time that deprotonated amino acid carboxylic group is able to change tautomeric state of a nucleotide base, exactly to convert the N9H ground-state prototropic tautomer of adenine into the N7H and N1H rare ones. PMID- 11392794 TI - [Prions and proteinaceous proteinase inhibitors: structural analogs and their consequences. II. Dynamics of prion diseases]. AB - The assumption about pathogenic prions as the proteins supplying the extracellular proteinases transport into intracellular space permits to bring the pathogenesis of prion diseases to order of the known and partially proved process regarding the case of prion diseases. We present the mathematical model of the dynamics of prion pathogenesis explaining the existence of the minimal infectious dose and small influence of its exceeding on the duration of long-term latent period of the disease. According to the model proposed the transformation of the neuronal cell into PrPSc breeder is the result of proteolytic damage of shaperoning system caused by accumulation in the cell of some crucial amount of proteinase-transporting prions. Such an accumulation is considered as the result of successive and centripheral lay-by-lay transformation of compact cellular locus from higher affinity to prions to normal one. The formation in the moveable frontier lays of the wave with high prion consisting and its closing into the locus center leads to dramatic splash of prion concentration even at moderate difference between higher and normal affinity levels. The final concentration of prions depends mainly on the correlation between these affinities whilst on exceeding of some value the dimension of the locus is of no importance. PMID- 11392795 TI - [Assessment of nitric oxide level in aqueous humor under physiological conditions and after lens extraction with PMMA during experimental work in rabbits]. AB - PURPOSE: To evaluate the presence of nitric oxide and measure its level in the aqueous humor of the rabbit's eye, in physiological conditions and after extracapsular lens extraction and PMMA artificial lens implantation. We also investigated nitric oxide maintenance during early postoperative period (between 1-5 day after surgery). MATERIAL AND METHODS: We used 30 rabbits (weighing 3.0 3.5 kg) Just before surgery samples of aqueous humor were aspirated by anterior chamber puncture. Lens was extracted with extracapsular (envelope) technique. In 15 eyes PMMA IOL was implanted in the bag and 15 eyes were left aphakic. The aqueous samples were collected on 1st, 3rd, 5th days after surgery. Nitric oxide in each sample was measured with respect to fluorometric assay. RESULTS: In aqueous humor in physiological conditions we detected nitric oxide. Its level was estimated on the value of 26.52 nM/dl. After extracapsular lens extraction in both groups the level of nitric oxide was significantly higher than in control group. The day and value of NO level was different among examined groups. Nitric oxide level diminished significantly on 5th postoperative day. CONCLUSION: We came to conclusion that after ECCE and PMMA IOL implantation NO level was significantly higher as compared with control. This higher NO level after lens extraction can be responsible for the blood aqueous breakdown. PMID- 11392796 TI - [Assessment of nitric oxide level in aqueous humor under physiological conditions and after lens extraction and artificial foldable acrylic lens implantation during experimental work in rabbits]. AB - PURPOSE: To evaluate the presence of nitric oxide (NO) and its level in the aqueous humor of rabbit's eye physiological conditions and after phacoemulsification and acrylic foldable artificial lens implantation. We also investigated nitric oxide maintenance during early postoperative period (between 1-5 days after surgery). MATERIAL AND METHODS: We examined 30 rabbits (weighing 3.0-3.5 kg) Just before surgery samples of aqueous humor were aspirated. Lens was extracted with phacoemulsification technique. In 15 eyes acrylic foldable IOL (group III) was implanted and 15 eyes were left aphakic (group IV). The aqueous samples were collected on 1, 3, 5 days after surgery. Nitric oxide in each sample was determined with fluorometric assay. RESULTS: The level of NO in aqueous humor in physiological conditions was estimated to 26.52 nM/dl. After phacoemulsification in both groups the level of nitric oxide was higher than in control group. The day and value of the highest NO level was different among examined groups. The highest level of NO was released during 1st day in group III and on the 3rd day in the group IV. CONCLUSION: We came to conclusion that the level of nitric oxide in aqueous humor after phacoemulsification in both groups is higher than in control group but significantly lower than in previously examined groups in which the surgery was made with extracapsular technique. PMID- 11392797 TI - [Isolation and culture of bovine choriocapillary endothelial cells using paramagnetic beads coated with Lycopersicon esculentum]. AB - PURPOSE: To establish a pure culture of choriocapillary endothelial cells as a model of angiogenesis in vitro. MATERIAL AND METHODS: Bovine choriocapillary endothelial cells (BCEC) were obtained by the method described by Hoffmann et al. (6) using the polystyrene paramagnetic beads coated with Lycopersicon esculentum, which attach specifically to the rest of fucose on the surface of microvascular endothelial cells. The endothelial characteristic of the cultured cells was evaluated by immunocytochemistry using anti von Willebrand factor and anti-CD 31 antibodies. Proliferation and survival of BCEC were tested using haemacytometer of Mallasez. RESULTS: The purity of obtained BCEC culture was confirmed by positive immunocytochemical staining with anti von Willebrand and anti factor CD 31 antibodies in more than 95% of cells. The proliferation of cells in Endothelial Cell Medium resulted in twofold increase of number of cells during 4 day observation period. After reaching the confluence, the cells continued to proliferate with increase of the cell number by 60% during 4-day observation. CONCLUSION: The use of paramagnetic beads coated with specific lectine provide a pure isolation of BCEC, which can be maintained in culture with preservation of their characteristic. PMID- 11392798 TI - [The effect of dopamine injected into the lateral brain ventricle on flash visual evoked potential (FVEP) after prenatal intoxication with cadmium]. AB - PURPOSE: Earlier studies demonstrated that exposure, especially prenatal, of mammalians to cadmium (Cd) results in disorders of visual evoked potentials (FVEP). The purpose of this study is to find out any influence of cadmium intoxications on the effect of dopamine (DA) on FVEP. METHOD: 18 Wistar albino rats in 3-6 months of age were divided into 3 groups: control (drinking tap water only), prenatal exposure of 5 ppm, and 50 ppm Cd. Flash visual evoked potentials (FVEP) were recorded before and after injections of dopamine (DA) 100 and 200 nmols into the lateral brain ventricle. The amplitude and latency of N1 and P2 were statistically analysed by the test of t-Student. p < 0.05 was used to indicate significant difference. RESULTS: There were no statistical differences of mean latency of P2 waves between initial records of FVEP and after both doses of DA in all observed groups. The amplitudes of both waves increased after DA injections compared to initial values in all groups. The differences were statistically significant. The mean latencies of N1 peak were prolonged after both DA doses (106-109%) compared to the initial records (100%) in the control group; however, there were no changes of it in the Cd-treated groups before and after DA injections. CONCLUSION: Cadmium blocked the dopamine effect on the latencies of N1 and P2 of FVEP (because it prolonged them alone); however, cadmium increased the stimulative effect of dopamine on their amplitudes. PMID- 11392799 TI - [Secondary glaucoma in patients with lens subluxation or luxation]. AB - AIM: To estimate the frequency of secondary glaucoma appearance and the influence of surgical treatment on intraocular pressure (IOP) in patients with posterior lens dislocation. MATERIAL AND METHODS: The study concerned 152 patients (57 females and 95 males, age range: 19 to 91 years, mean--58 years). Secondary glaucoma was found in 62 patients (41%). Trauma was the cause of lens dislocation in 81% of these cases, while it was the cause in only 69% of cases in the whole material. Pars plana vitrectomy with limbal intracapsular lens extraction or lentectomy were performed in all cases. In 24 cases perfluorocarbon liquid was used. Scleral fixation PC IOLs were implanted in 85 eyes (group A), AC IOLs in 27 eyes (group B), and 40 eyes remained aphakic (group C). The follow-up time ranged from 6 months to 5 years, mean 22 months. RESULTS: Raised IOP was found preoperatively in 46 cases (30.3%). It was raised in 21 (24.7%) eyes in group A (24-60 mm Hg, mean 37.1 mm Hg), and it has normalized postoperatively in 14 eyes, while topical medication was necessary in 7 remaining cases. In group B raised IOP (26-60 mm Hg, mean 41.4 mm Hg) was found preoperatively in 8 (29.6%) eyes. It has normalized postoperatively in 5 eyes, while topical medication was necessary in 3 eyes. The respective values in group C were: 17 (42.5%), 24-80 mm Hg, mean 43.2 mm Hg. In 7 eyes IOP was normal after surgery, 10 patients required topical treatment, and in 3 of these cases trabeculectomy had to be performed. In 16 eyes with normal preoperative IOP it has raised postoperatively in different periods of time (from 1 week to 6 months). CONCLUSIONS: Secondary glaucoma is a frequent complication of the posterior lens dislocation, and it is more often related with traumatic cases. The dislocated lens removal with the use of vitrectomy causes IOP normalization in most of the cases. The periodical examination of IOP is necessary after the dislocated lens removal also in cases with normal preoperative IOP, because secondary glaucoma may appear in different time after surgery. PMID- 11392800 TI - [Intraocular correction of aphakia after cataract extraction for uveitis in adolescents]. AB - PURPOSE: To demonstrate 3 cases with pediatric pseudophakia after cataract extraction as a complication of unilateral uveitis. MATERIAL AND METHODS: The boys were at the age of 8, 11 and 15 years at the moment of surgery. Uveitis was monocular, the etiology unknown and clinically significant cataract was diagnosed after several months from the beginning of inflammation. The last recurrence of uveitis was observed 1 to 6 months before operation. There were intermediate uveitis (2 cases) and anterior uveitis (1 cases). In 2 eyes PMMA implant was placed in-the-bag and in one eye in the ciliary sulcus. During surgery we did not observe any complications except greater tendency to bleeding. Follow-up after operation ranged from 1.5 to 2.5 years. RESULTS: Visual acuity before cataract extraction was 0.01; 0.04 and 0.02 and during last visit after surgery it was 0.9, 0.6 and 0.4 respectively. Postoperative near distance visual acuity was 0.5, 0.75 and 1.25. We observed cystoid macular oedema in the latter case. Two months after PC-IOL implantation the symptoms of mild inflammation in the eye of 15-year old boy with anterior uveitis occurred. Intraocular pressure before and after operation ranged from 12 to 17 mm Hg. Corneal central endothelial cell density was from 2850/mm2 to 3100/mm2 before and from 2469/mm2 to 2979/mm2 during last visit. CONCLUSION: Cases of posterior pseudophakia during uveitis in children showing good functional and anatomical state of eyes in the long term observation after IOL implantation recommend the intraocular correction in carefully selected uveitic pediatric patients. PMID- 11392801 TI - [Analysis of glaucoma therapy costs in Poland. I--population of the Mazovian District]. AB - AIM OF THE STUDY: The purpose of this study was to estimate and evaluate costs of glaucoma therapy in Poland in 1998-2000 years. MATERIAL AND METHODS: 315 patients (591 eyes) including 364 POAG eyes and 227 PACG eyes were evaluated. Progression of the disease was measured in two modes: by changes in the optical nerve disc (c/d) and by IOP level. Duration of the study was 12 months. Total costs (e.g. number of visits, amount of medications used, surgical and laser procedures performed) were calculated on 100 eyes/12 months. RESULTS: Pharmacological therapy constitutes 40-50% of the total costs in both glaucoma types. Timolol was the mostly prescribed drug in POAG, while pilocarpine led in PACG group. Dorzolamide was on the third place, but it generated the biggest part of total costs. Patients with advanced forms of the disease more frequently saw doctor on visits (average 6 visits per year); patients with elevated IOP (> 21 mm Hg) underwent surgical procedures more commonly than these with optical nerve disc changes (c/d > 0.6). The total costs of the therapy of non-advanced cases amounted 70.243 PLN on the average, while in advanced cases it reached the average of 155.230 PLN. CONCLUSION: 1. The total cost of glaucoma therapy depends on progression of the disease. 2. The most cost-generating part is the pharmacological therapy of glaucoma. PMID- 11392802 TI - [Methods for in vivo measurement of light transparency in the human crystalline lens]. AB - A method for the in vivo measurement of light transparency of the human crystalline lens, based on the measurement of the blue and red light threshold radiance, in presented. The threshold limit values for radiances of 477 nm (blue) and 620 nm (red) were measured for people at age from 17 to 66 years old without cataract. The people recognised the geometric figures by left and right eye separately, after being adapted to darkness. The radiance of the figures was changed smoothly by ADM adaptometer. The threshold limit values were measured several times for each eye. For each eye the average of the threshold limit values for radiances of 620 nm and 477 nm and their quotient were calculated. Values of these quotients practically did not change for young eyes (to 32 years old) and they decreased for older eyes. These results indicate the increase of the threshold radiance of 477 nm in relation to threshold radiance of 620 nm for older people, and this means the reduction of the crystalline lens transparency for blue light in relation to red one as they grow old. It seems that the reduction should precede the development of cataractous changes and this fact may be used in the diagnostics of precataractous states. PMID- 11392803 TI - [Cataract surgery in adults and children with chronic uveitis]. AB - PURPOSE: To present preoperative strategies and results of cataract surgery in patients with different types of uveitis. PATIENTS AND METHODS: The results of 18 cases of extracapsular cataract extraction with or without intraocular lens implantation were retrospectively analysed. Age of patients ranged from 10 to 71 years (mean age: 35 years). Etiology of uveitis was determined in 10 cases (63%). Preoperative management was determined by the diagnosis of uveitis. The postoperative observation time ranged from 3 years to 6 months (mean: 25 months). RESULTS: Visual acuity improved postoperatively in 17 eyes (96%), and was better than 0.5 in 10 eyes (63%). The reasons for lower visual acuity were: cystoid macular edema in 6 cases (33%), band keratopathy, amblyopia and floaters in the vitreous. CONCLUSIONS: Establishing the diagnosis and controlling the inflammation pre- and postoperatively are the basic conditions for the surgical outcomes in cataract surgery in patients with history of uveitis. PMID- 11392804 TI - [Observation of the cornea with confocal microscopy after Nd:YAG laser capsulotomy]. AB - PURPOSE: The purpose of the study was to evaluate the influence of Nd:YAG capsulotomy on the cornea structure assessed by scanning slit confocal microscopy. MATERIAL AND METHODS: 14 eyes in 14 patients after Nd:YAG laser capsulotomy performed 6-34 months after extracapsular cataract extraction with PCIOL implantation were observed. Structure of cornea layers was evaluated before the procedure, then 3 hours and 7 days after the laser exposition using confocal microscope. RESULTS: Initial evaluation showed degenerative changes in subepithelial plexus; 3 hours after the procedure excessive exfoliation of superficial epithelial cells was observed. No significant changes were registered 7 days after the procedure. CONCLUSION: No significant changes were found in the confocal microscopy images of cornea after Nd:YAG capsulotomy. PMID- 11392805 TI - [Causes of tilt in the posterior chamber lens around the axis of the suture following its attachment to the sclera]. AB - PURPOSE: The aim of the study is to find out how the way of tying the suture around the haptics creates the tilt of the lens around the axis of the suture. MATERIAL AND METHODS: The study was carried out using a transparent polyethylene cylinder 30 cm long 12 mm in diameter similar to the one of the eyeball in the area of sulcus ciliaris. The lenses were sutured about 5 mm from one of the ends of the cylinder using 9-0 suture. The deviation of the lens was evaluated be measuring the distance of the circumference of the haptic from the edge of the cylinder. The study was carried out using lenses without fixation holes in loops with the holes in one loop and holes in both loops. Various ways of tying the suture in the holes of the haptic were applied. RESULTS: The lenses without holes in loops were tilting around the axis of the suture. Stabilisation of these lenses was received after using two sutures on one of the loops. Similarly, lenses with hole in one loop were not tilting just after using two sutures in loop without hole. The lenses with two holes in loops were not tilting when fixation sutures were tying in a way which balances the tilt forces of sutures on each loop. CONCLUSIONS: A kind of tying the suture around the haptic has significant importance on tilt of the lens around the axis of the suture. In case of necessity to suture lenses without holes in the haptics three or four sutures (two or one on each haptic) should be applied and they must be pierced in the sclera at the distance of 5 mm from each other. Lenses with hole on one loop should have two fixation sutures on loop without hole. Lenses with holes in two loops should have sutures tying in a way which balances the tilt forces of sutures on each loop. PMID- 11392806 TI - [Eight year results of excimer laser correction for irregular astigmatism by linear excimer laser photoablations of the cornea in cases of keratoconus]. AB - PURPOSE: The purpose of this study was to evaluate changes of the corneal curvature as a result of linear incisions with excimer laser and also to evaluate durability of the achieved effect, including the changes in visual acuity, topographic maps of the corneal curvature and keratometric changes at the location of keratoconus 2, 4, 8, 16, 24 weeks postoperatively. MATERIAL AND METHODS: At the First Department and Clinic of Ophthalmology, Silesian Medical School in Katowice, 294 patients went through ophthalmological examinations. 164 patients (187 eyes) with the III degree of keratoconus according to the Amsler scale, including 69 women (77 eyes) and 95 men (110 eyes), were classified for the excimer laser surgery with the method of linear, corneal incisions with MEL 60 (Aesculap-Meditec). The eyes did not tolerate contact lenses at all or because of peripheral location of the keratoconus. The patients were from 16 to 60 years of age (the mean age was 27.9 years). RESULTS: According to the statistical results, there were no difference between the effects of treatment between men and women. Correlation between the results of the surgery and the patients age showed that the latter did not have any effect on the final result. Lack of correlation between the height of the absolute corneal curvature at the keratoconus and the achieved change after the surgery was reported, thus changes of the corneal curvature parameters do not depend on its parameters before the surgical procedure. Absolute visual acuity of patients was improving and reached maximally 0.252 of the unit. As a result of the single surgery, the maximum change of keratometric values was 4.949 D, and the minimum was 3.611 D. In the case of the secondary surgery, the maximum keratometric value at the axis of the incision was 8.55 D and the minimum one was 7.101 D. CONCLUSION: To sum up, we can say that the favourable effect of decreasing the irregular corneal astigmatism with the method of linear laser fotoablations was maintained throughout the whole observation period, which allowed to shift in time planned corneal transplantation. PMID- 11392807 TI - [Correction of irregular astigmatism using excimer laser MEL 70 G-Scan with the TOSCA program--introductory report]. AB - PURPOSE: The purpose of this study was to evaluate changes of the corneal curvature as a result of excimer laser ablation using the TOSCA and TSA system. MATERIAL AND METHODS: At the First Department and Clinic of Ophthalmology, Silesian Medical School in Katowice, 8 patients (8 eyes) with the II and III degree of keratoconus according to the Amsler scale, including 4 women (4 eyes) and 4 men (4 eyes), were classified for the excimer laser surgery with the method of PRK with TOSCA program and TSA system with MEL 70 G-Scan (Asclepion-Meditec). The eyes did not tolerate contact lenses at all or because of peripheral location of the keratoconus. The patients were from 18 to 42 years of age (the mean age was 25.8 years). The observation period was 4 weeks. RESULTS: According to the statistical results lack of correlation between the height of the absolute corneal curvature at the keratoconus and the achieved change after the surgery was reported. The improvement of patients absolute visual acuity was improving and reached 0.2 of the unit. CONCLUSION: To sum up, we can say that the favourable effect of decreasing the irregular corneal astigmatism with the method of laser fotoablations with topography-controlled system was maintained throughout the whole observation period, which probably allowed to shift in time planned corneal transplantation. PMID- 11392808 TI - [Risk factors of 3rd stage retinopathy of prematurity progression]. AB - Retinopathy of prematurity is a vasoproliferative, multifactorial disease of retina affecting the preterm neonates. Purpose of the study was to assess the association between a range of clinical features and the development of the stage 3 ROP. The studies comprised 79 preterm infants with diagnosed 3rd stage of ROP and 1117 preterm neonates without ROP as a control group. The analysis included the clinical data of patients--gestational age, birth weight, presence of hypotrophy, Apgar score, duration of oxygen therapy, ventilation and phototherapy. Among the potential risk factors, hypotrophy and duration of phototherapy had no significant influence on the development of 3rd stage ROP. High risk of development of 3rd stage ROP was observed in neonates born before 28th week of gestation, weighing at birth less than 1000 g. The significant risk factors were also the asphyxia, duration of ventillation exceeding 30 days and oxygen therapy longer than 40 days. PMID- 11392809 TI - [Grave's and Basedow's thyroid associated ophthalmopathy--pathogenesis, symptoms and selected aspects of treatment]. AB - The author presents the review of new literature concerning advances in therapy, especially immunosuppressive, orbital radiotherapy, as well as pathogenesis, epidemiology, symptoms and natural history of thyroid associated Graves' ophthalmopathy. PMID- 11392810 TI - The treatment of corneal ulceration. PMID- 11392811 TI - [Interzonal microtubules and meiotic restitution in the dycots]. AB - Dynamics of the microtubule cytoskeleton at mobile stages of female meiosis in the dycots was studied. A new stage of MT rearrangement was revealed referred to as centrifugal movement of interzone MTs at telophase 1. A disruption of this process leads to the displacement of daughter nuclei to the equatorial region and a common (fused) spindle formation. This is the cause of dyad (instead of tetrads) formation in mutant ps of Beta vulgaris. PMID- 11392812 TI - [Morphogenesis and pattering in hydra. II. Molecular mechanisms]. AB - Molecular mechanisms of morphogenesis and pattern formation have been extensively studied during the last decades. Recent data suggest that many of the signalling systems as well as transcription factors governing embryonic development in the higher animals have been already established in the lower Metazoa. This review summarizes the information on the roles of peptide systems, signal transduction cascades, transcriptional activators and the extracellular matrix-processing enzymes in the developmental processes in Hydra. PMID- 11392813 TI - [Stability of spatial interactions between chromocenters and pre-kinetochores in the interphase murine cells]. AB - It is known that in mice the centromeric heterochromatin remains compact during the whole cell cycle and at interphase is referred to as "chromocentres". In the current study, by the use of antibodies against prekinetochores and DNA polymerase (a PCNA antigen), we showed that in murine L929 cells chromocentres remain spatially associated with prekinetochores during the entire interphase, including the late S-period, when DNA chromocentres replicate. Augmentation of prekinetochore fluorescence increases concomitantly with the heterochromation replication, but the prekinetochore duplication occurs only in G2 period. A conclusion has been made that murine interphase cells can be used for biochemical fractionation of chromocentres associated with prekinetochore proteins. PMID- 11392814 TI - [Mechanisms of Ca2+ signalling in cells]. AB - The review summarizes recent data and current opinions of the Ca2+ signal formation in cells. Mechanisms of Ca2+ mobilization from the intracellular Ca2+ stores are discussed along with the pathways of Ca2+ entry from the external medium. PMID- 11392815 TI - [The murine nuclear matrix protein specificcaly binds to centromeric satellite DNA]. AB - The nuclear matrix (NM) of mouse contains a protein (miSat BP) that can specifically bind to mouse centromeric minor satellite DNA as shown by gel shift assay. The ion-exchange chromatography on DEAE-Sepharose was used as the first miSat BP purification. MiSat BT was eluted by 0.2 M NaCl. Antibodies against p70, a human NM protein of 70 kDa described earlier as a protein recognizing human alphoid DNA, produce hypershift effect when added to the retardation incubation mix. Immunoblotting of NM and an active NM fraction (0.2 M NaCl) with these antibodies revealed a protein with 70 kDa in both preparations. This antigen retained in NM in situ being associated with residual DNA as shown by indirect immunofluorescent staining. In the untreated interphase nucleus most of miSat BP granules were shown to be colocalized with prekinetochores. We suggest that miSat BP is capable of recognizing the minor satellite DNA due to its structural features, but it does not belong to a group of constitutive centromeric proteins. PMID- 11392817 TI - [Stress-induced DNA fragmentation in cardiomyocytes of MDX and C57Bl mice]. AB - The study deals with apoptosis of cardiomyocytes of adult mdx mice. Our previous investigation of the total DNA from mdx mice myocardium in 0.5% agarose and 5% PAAG revealed middle sized DNA fragments about 64 kb only. The DNA electrophoresis in 5% PAAG after alpha-32P-dATP incorporation into DNA, fragments mediated by terminal transferase, showed no presence of smaller DNA fragments. The low size DNA fragments (0.2, 0.4 and 11.0 kb) were observed only following stress (a 5 min swimming in water bath) by means of alpha-32P-dATP incorporation mediated by TdT reaction. DNA fragmentation lasted for 48 h after the stress. A study of the total DNA extracts of C57B1 mice also demonstrated middle and low molecular sized DNA fragmentation 2 h after the stress under the same condition of the experiment. But during the next 48 h the signs of DNA fragmentation of the total DNA disappeared. The authors conclude that the first stage of apoptosis is a permanent property of cardiomyocytes of mdx mice. PMID- 11392816 TI - [The role of cytoskeleton structures in regulation of Ca2+ responses in macrophages]. AB - The role of the cytoskeleton in regulation of purinergic agonist- and endoplasmic Ca(2+)-ATPase inhibitors-induced Ca2+ signals in rat peritoneal macrophages was investigated. It has been shown that in cells pretreated with agents that disrupt microtubules (vinblastine, colchicine, colcemid) or actin microfilaments (cytochalasins, phalloidin), the ability of thapsigargin or cyclopiazonic acid to empty Ca2+ stores and activate store-dependent Ca2+ influx was significantly attenuated. On the contrary, microfilaments and microtubule disrupters did not affect ATP- or UTP-induced Ca2+ mobilization, indicating that release of Ca2+ from intracellular stores through the inositol phosphate pathway was intact. The results suggested that an intact cytoskeleton is required for capacitative Ca2+ entry but not for agonist-induced Ca2+ mobilization. PMID- 11392819 TI - [The effect of metabotropic glutamate receptors on longitude of posttetanic reaction in spinal motoneurons of frog]. AB - Effects of metabotropic glutamate receptors of the duration of posttetanic changes in monosynaptic excitatory postsynaptic potentials (mEPSP), evoked by afferent and reticulospinal input stimulation, were investigated in lumbar motoneurons of the frog isolated spinal cord. It was found that application of MAP4 (25 microM), a selective antagonist of group III of these receptors, prolonged posttetanic potentiation and depression of synaptic transmission, whereas activation of this group of metabotropic glutamate receptors by L-AP4 (1 mM), a selective agonist of these receptors, suppressed the amplitude of synaptic responses, but did not affect the dynamics of development of posttetanic changes. The NMDA receptor antagonist AP5 (50 microM), added to the perfusing solution, blocked completely the effects produced by MAP4. Neither selective antagonist MCCG (400 microM), nor agonist tACPD (50 microM) of group II metabotropic glutamate receptors affected the terms of mEPSP posttetanic potentiation and depression, although the latter, in contrast to the antagonist, in most cases increased the synaptic potential amplitude. The data obtained permit to suggest that group III metabotropic receptors may control the duration of posttetanic changes of synaptic transmission in the frog spinal motoneurons. The long-term changes in the investigated synapses seem to be mediated by activation of postsynaptic metabotropic glutamate receptors (most likely, of group I receptors), which is normally masked with activation of group III presynaptic autoreceptors. The mechanism of such an induction essentially depends on activation of NMDA type of inotropic glutamate receptors. PMID- 11392818 TI - [Combination of thymidine and dexamethasone increases the K562 tumor cells sensitivity to human leukocytes cytotoxicity]. AB - The influence of the number of differentiating agents on sensitivity of human erythroleukemic cells K562 to human leukocyte-mediated non-MHC-restricted lysis was studied. It has been shown that a 4-day treatment of cells K562 by dexamethasone (1 microM) or phorbol-12-myristate-13-acetate (100 nM) leads to a significant decrease in sensitivity of the treated cells to non-specific lysis mediated by human leukocytes. On the contrary, the treatment of cells K562 by a combination of dexamethasone and thymidine (2 mM) leads to an increased sensitivity of the treated cancer cells to non-specific lysis mediated by the above effector cells, compared with the situation when these cells were treated by dexamethasone only. The treatment of cells K562 by a combination of thymidine and phorbol-12-myristate-13-acetate demonstrates a tendency (P < 0.1) to increase the sensitivity to non-specific lysis mediated by human leukocytes, as compared with the cases, when these cells were treated by phorbol ester only. It has been shown that the changes in K562 cell sensitivity to lytic action of leukocytes, under the chosen incubation time and doses of the used agents, well compare with the changes of erythroid differentiation of the cancer cells in the same conditions. PMID- 11392820 TI - [Immunohistochemical study of cell phenotypes in organ culture of the rat embryo liver]. AB - The organ culture of the liver of a 14 day old rat embryo was used for immunohistochemical recognition of the available cell types. This culture contains the tissue explant, confluent monolayer cells and a zone of single cells (ZSC). At the start of cultivation a monolayer is formed at the expense of cells slipping down together from the explant, and later these cells were seen to migrate actively from the explant and to proliferate. ZSC is formed by cells migrating from the monolayer. Desmin-containing cells (hepatoblasts, Ito cells and myofibroblasts) migrate from the explant to the monolayer and ZSC. In the monolayer hepatoblasts lose gradually desmin and are converted into cytokeratin 18 (CK-18) expressing hepatocytes. In ZSC, hepatoblasts lose desmin, but no CK-18 synthesis occurs in them. The Ito cells (hypothetical progenitor cells) are spreading over the whole culture, and desmin expression in them does not change. The embryonal Ito cells may transform presumably into myofibrils. Myofibroblasts lie flat on the periphery of ZSC. Besides, desmin myofibroblasts express alpha actin from smooth muscles. Expression of CK-18 in cells depends on the pattern of intercellular interactions. In the monolayer, CK-18 expression extends successively via the adjacent cells towards the explant. In hepatocytes, migrating into ZSC, CK-18 expression stops. In ZSC, CK-18 expression reappears in compact clusters of stopped cells. It supposed that during formation of close contacts in the monolayer the cells-predecessors may be differentiated into hepatocytes, whereas in the case of disturbance of intercellular connections in ZSC the cells-progenitors may be converted into myofibroblasts. However, on the reconstruction of cell contacts in motionless dense clusters the cells in ZSC are differentiated into hepatocytes. PMID- 11392821 TI - A descriptive analysis of the use and cost of new-generation antihistamines in the treatment of allergic rhinitis: a retrospective database analysis. AB - OBJECTIVE: This retrospective database analysis was conducted to evaluate the use and cost of new-generation antihistamines (i.e., those that are nonsedating) in the treatment of allergic rhinitis in a managed care population. STUDY DESIGN: The study is a retrospective database review of medical and pharmacy-related claims linked by episodes of care. METHODS: Patients who had been diagnosed as having allergic rhinitis and had at least 1 prescription claim were identified from a database containing patient-level medical and pharmacy-related claims. The treatment patterns of patients with allergic rhinitis who met the study criteria were documented for a 12-month period in which the use of nonsedating antihistamines was described and the associated costs of various medications were assessed. Subanalyses of patients categorized by comorbidity status were also performed. RESULTS: A total of 202,426 patients participated in the study. Nonsedating antihistamines were used by 71% of the patients; the most commonly prescribed drugs were loratadine and fexofenadine. The mean annual charges per patient for the treatment of allergic rhinitis in the study population were $465.21 (standard deviation [SD], 548). The greatest departmental cost was that of pharmacy-related charges (mean, $236.02; SD, 233); the next highest cost was that of outpatient charges (mean, $216.31; SD, 396). Comparisons of departmental charges indicated the use of loratadine was associated with significantly higher treatment costs than that of fexofenadine in a number of patient subgroups. CONCLUSION: In this analysis, loratadine was associated with significantly higher treatment charges than was fexofenadine. This result was observed consistently across different stratifications of patients, including the presence of comorbid respiratory infection, concomitant use of nasal steroids, and the presence of asthma and/or sinusitis. These results provided useful insights into the differential costs associated with the use of nonsedating antihistamines in the treatment of rhinitis. PMID- 11392822 TI - [Exophthalmia]. AB - The authors present the association of signs and symptoms in exophthalmos syndrome, which occurs in many diseases. Although exophthalmos was considered to be of ophthalmological interest, according as researches were made on its pathogeny and according as investigation methods were developed, exophthalmos is no more considered a symptom, but a syndrome. After defining the syndrome, the present study shows its classifications, clinical and complementary investigations, and its treatments. PMID- 11392823 TI - [Open angle glaucoma. Identification of vascular risk factors]. AB - Although the clinical picture of primary open-angle glaucoma (POAG) is well described, the exact mechanism leading to this specific type of damage is still under investigation. Among the vascular risk factors, hypotension and vasospasm seem to have an important role in the pathogenesis of this disease. This review presents the importance of hypotension and vasospasm in the pathogenesis of POAG and an inventary of the principal methods for their detection. PMID- 11392824 TI - [Etiopathogenic consideration in the development of retinal detachment in aphakic and pseudoaphakic eye]. AB - PURPOSE: Is to evaluate the risk factors implicated in the development of retinal detachment in aphakic and pseudophakic eyes. MATERIAL AND METHOD: We studied 46 cases operated for cataract by planned extracapsular cataract extraction with or without intraocular lens, wich developed after the operation retinal detachments. RESULTS: Retinal detachment appeared in 2.98% cases of EEC with AC-IOL and in 0.56% in EEC with PC-IOL. We performed Yag laser capsulotomy in 4 cases. Retinal detachment appeared in 12 cases between 6-12 months. The most frequent breaks causing retinal detachment where "horse shoe tears" in 23.4% of cases and tears in 14.89% cases. CONCLUSIONS: 1. High myopia and peripheral retinal degenerations are risk factors in the development of retinal detachment after the cataract operation. 2. Vitreous loss facilitates the appearing of vitreo-retinal tractions followed by retinal detachment. 3. PC-IOL reduces the frequency of retinal detachment by the stabilisation of the eye and limiting the ophthalmodonesis. PMID- 11392825 TI - [About residents and residency]. PMID- 11392826 TI - [The influence of meteorological factors for frequency of Herpes simplex keratitis]. AB - HYPOTHESIS: The variation of the meteorological factors could be a stress for the organism, which could determine the start of some diseases. MATERIAL AND METHOD: We studied 252 cases of herpes simplex keratitis which had been hospitalized in Eye Clinic of Cluj between 1983-1991. The selection criteria were: known diagnosis; day of the rise could be exactly established; disease was taken place in Cluj or in region not longer than 30 km on Somes Valley. The meteorological parameters were collected by specialist from the Meteorological Station of the city and from the Cluj's Airport. The results were verified statistical and were guaranted by p = 0.05. RESULTS: The herpes simplex keratitis are more frequently in days with a temperature higher and an atmospheric pressure lower than the average value of the season. DISCUSSIONS: We drew a possible pathogenic mechanism to explained the reactivation of the virus under the influence of warm weather. PMID- 11392827 TI - [Association of cataract and glaucoma. Therapeutical attitude]. PMID- 11392828 TI - [Ocular refraction and branch of retinal vein occlusion]. AB - PURPOSE: To evaluate the significance of refractive errors in cases of branch retinal vein occlusion (BRVO). METHODS: We included in this study 179 patients with BRVO who attended our department between 1996-2000. We compared this group with an equal number of age, sex and associated diseases matched controls. The spherical equivalents of the refractive errors of patients in both groups were compared using the chi-square test and Student's t test. RESULTS: Hypermetropia was present in 114 patients wit BRVO (63.68%) and in 82 controls (45.8%, p = 0.00048). Myopia was present in 16 patient with BRVO (8.9%) and in 39 controls (21.8%, p = 0.0037). Emetropia was found in 49 patients (27.37%) and in 58 controls (32.4%, p = 0.299). CONCLUSIONS: Hypermetropia is significantly more common in patients with BRVO than in general population, but myopia is significantly less common in these patients than in controls. PMID- 11392829 TI - [Immunology and immunopathology of corneal graft]. AB - The article shows the immune reaction against an allograft, specially against a corneal graft. Then are shown the mechanism of graft rejection and the posibilities of immunologic treatment of this complication of keratoplasty. PMID- 11392830 TI - [Postoperative induced astigmatism]. AB - PURPOSE: Is to evaluate the astigmatism induced by cataract surgery, ethiological factors and ways of prophilaxies and treatment. MATERIAL AND METHOD: We followed a number of 211 eyes operated for cataract by planned extracapsular extraction with intraocular lens implantation, in Ophthalmological Clinic from Cluj-Napoca. All the pacients had limbal incision and the suture was performed with 9.0 suture, after the operation the treatment consisted in local drops with steroids. REZULTS: The astigmatism induced by the surgery was direct in 53.08% of cases and indirect in 46.92%. The position of the axes was 0-900 in 72.51% and oblique in 27.48%. The optical correction of astigmatism was with combined spherocylinder glases in 57.34% and only with cilinder in 42.65% of cases. CONCLUSIONS: 1. The induced astigmatism by cataract surgery is in relation with the incision, suture and IOL. 2. Correction of astigmatism is possible with optical correction, repearing the suture or with refractive surgery. 3. The prevention of astigmatism over 1.5 D is by using Troutmann keratometer and to evidence the congenital astigmatism. PMID- 11392831 TI - [Actualities in retinoblastoma's treatment]. AB - The management of retinoblastoma should be undertaken by multidisciplinary health care team. Ophthalmopediatrics and ocular oncologists have sought effective alternative methods for treating retinoblastoma. The management of retinoblastoma has been changed significant past few years. Recently, advances in understanding of retinoblastoma have led to trials of new treatment modalities aimed at decreasing morbidity and continuing excellent survival. This review tries to present the new and modern aspects in retinoblastoma therapy. PMID- 11392832 TI - [Combined surgical procedures in strabismus]. AB - PURPOSE: This is a study about the simultaneous horizon-to-vertical surgery and about the myopexia associated with the horizontal rectus muscles surgery. The aim of this article is to demonstrate necessity of these simultaneous operations into different etiopathogenetical strabismus forms. METHODS: A number of 30 patients were included in the first lot. In this cases we used a simultaneous horizontal and vertical rectus muscles surgery interventions. The second lot--163 patients- has been operated through classical methods on rectus associated with myopexia. RESULTS AND CONCLUSIONS: The action on the etiopathogenetical factors of strabismus explains this favorable results (more than 80% after 6 months) obtained through combinated surgery intervetions. That is why this operations are indicated in any strabismus forms. PMID- 11392833 TI - [Original method for IOL sulcus fixation (a quasi-closed technique)]. PMID- 11392834 TI - [Phacoemulsification and foldable IOL implantation; posterior luxation of the nucleus; delayed phacophagia]. AB - In the beginning, the paper presents some general words about the main intraoperative complications of the phacoemulsification and some practical attitudes patient in front of the rupture of the posterior capsule. The second part presents the case of a 63 years old patient who was intended to be operated for cataract by phacoemulsification and who suffered a complication during this procedure--the rupture of the posterior capsule with the luxation of the majority of the nucleus into the vitreous body. PMID- 11392835 TI - [Endo-ocular surgery in uveitis. Case report]. PMID- 11392836 TI - [Total rhegmatogenous retinal detachment - late postoperative complication]. AB - This paper presents the case of a pacient operated for organized hemoftalmus after central retinal vein thrombosis (total vitrectomy) and whom five days after surgery a macular hole was observed and internal indendation with gas was performed. Postoperative evolution was good. Two months after surgery the pacient returns with cvasitotal regmatogen retinal detachment. The possible causes in bringing about of the retinal detachment in this case will be discussed as well as the factors that condition the success in posterior pole surgery. PMID- 11392837 TI - [Difficulties of diagnosis in a case of optic nerve drusen]. AB - This paper presents a particullary case of optic nervue drusen. Ophthalmoscopic features (superficial drusen in one eye and deep drusen in another), clinical association with anisometropia and ambliopia, wrong interpretation of PEV and tomodensitometry mod to diagnostic confusion, supplementary investigations, prolonged and expensive treatment. PMID- 11392838 TI - [Succesive metastasis in the right orbit with starting point a skin malignant melanoma located under the right clavicle]. AB - It's presented a case of the successive metastases in the right orbit with starting point a malignant melanoma skin located under the right clavicle. It's discussed the raruty situation of the malignant melanomas skin metastases in the orbit. PMID- 11392839 TI - Coverage and access. Insurance makes a heartland comeback. PMID- 11392840 TI - Real estate. Sticker shock for some California docs. PMID- 11392841 TI - Nursing. Overtime at issue. PMID- 11392842 TI - Diagnostic imaging. Scan on demand. PMID- 11392843 TI - Telecommunications. Everywhere at once. PMID- 11392844 TI - The leapfrog effect. AB - Some of the nation's largest health care purchasers, including such corporations as General Motors, UPS, General Mills and Xerox, are trying to exert more influence over patient safety programs. Hospitals and other providers are wary about the impact that the Leapfrog Group and similar employer organizations will have on health care. PMID- 11392845 TI - No easy answers. AB - Illegal immigration is on the rise and Congress has restricted access to health coverage for legal immigrants. Those factors combine to create a large pool of uninsured patients, and hospitals once again are left to foot the bill--to the tune of millions of dollars a year. And the problem is no longer restricted to border states and big cities. PMID- 11392846 TI - [Receptor occupancy and antipsychotic drug action measured by PET and SPECT]. AB - Positron emission tomography (PET) and single photon emission computed tomography (SPECT) studies have demonstrated consistent findings of high dopamine D2 receptor occupancy (> 65-70%) in patients treated with antipsychotic drugs. Further, the risk of extrapyramidal side effects has been shown high in patients with occupancy above 80%. On the basis of these findings, an optimal interval for D2 receptor occupancy between 70% and 80% has been suggested. It has also been shown that several atypical antipsychotics induce marked occupancy of central 5 HT2 and D2 receptors in vivo. However, a low D2 occupancy has been observed in patients with clinical dose of clozapine or quetiapine. The antipsychotic effect of these atypical drugs with a low D2 receptor occupancy has been widely discussed with respect to actions on other receptor systems, limbic selectivity of antipsychotic action and episodic transient occupancy. The recent advances in PET/SPECT and developments of new radioligands have made it possible to evaluate antipsychotic drug actions directly in humans. The empirical data from occupancy measurements will enable us to open future directions of investigation of antipsychotic action and improvement of antipsychotic treatment. PMID- 11392847 TI - [Investigation of verdicts of civil suits brought against hospitals involving crimes committed by mental hospital inpatients with psychotic disorders]. AB - The author investigated 11 verdicts of Japanese civil suits brought against hospitals involving crimes committed by mental hospital inpatients with psychotic disorders, principally by victims of injuries or by surviving relatives of homicide victims. About the half of the verdicts are based on the same logic. First, a detailed, case-specific investigation was performed. In some cases it was proven that the accidents could not have been predicted, and the suits were dismissed. In others the plaintiffs won the suits, because it was found that the violent acts were predictable and that the hospital staff had the means to prevent them. These verdicts require the hospital staff to predict the patient's actions according to the psychiatric history and present condition and then to make an appropriate response. This process is similar to the clinical decision making process, although there have been some dubious findings in some of the court cases. However, the author also found that some verdicts were inappropriate and insufficient in that there was no individual investigation of the offender. Examining all 11 verdicts, the author found that the type of admission procedure adopted for the offender, whether they had a past history of violence, and whether the hospital was public or private did not seem to be significant to the judges in making their decisions. PMID- 11392848 TI - [Some questions to Dr. Takigawa's article (Part II)]. PMID- 11392849 TI - [Answer to Dr. Kato's article (Part II)]. PMID- 11392850 TI - Physicians fight expansion of scope of nursing practice. PMID- 11392851 TI - Cyberspace and medical records? PMID- 11392852 TI - Intranasal preparations. PMID- 11392853 TI - House outlines tasks for 2000. PMID- 11392854 TI - KePRO priming for adult vaccination campaign. PMID- 11392855 TI - Emotional states and physical health. AB - Positive emotional states may promote healthy perceptions, beliefs, and physical well-being itself. To explore potential mechanisms linking pleasant feelings and good health, the authors consider several lines of research, including (a) direct effects of positive affect on physiology, especially the immune system, (b) the information value of emotional experiences, (c) the psychological resources engendered by positive feeling states, (d) the ways in which mood can motivate health-relevant behaviors, and (e) the elicitation of social support. As anticipated by the Greek physician Hippocrates, positive emotions and healthy outcomes may be linked through multiple pathways. PMID- 11392856 TI - Wisdom. A metaheuristic (pragmatic) to orchestrate mind and virtue toward excellence. AB - The primary focus of this article is on the presentation of wisdom research conducted under the heading of the Berlin wisdom paradigm. Informed by a cultural historical analysis, wisdom in this paradigm is defined as an expert knowledge system concerning the fundamental pragmatics of life. These include knowledge and judgment about the meaning and conduct of life and the orchestration of human development toward excellence while attending conjointly to personal and collective well-being. Measurement includes think-aloud protocols concerning various problems of life associated with life planning, life management, and life review. Responses are evaluated with reference to a family of 5 criteria: rich factual and procedural knowledge, lifespan contextualism, relativism of values and life priorities, and recognition and management of uncertainty. A series of studies is reported that aim to describe, explain, and optimize wisdom. The authors conclude with a new theoretical perspective that characterizes wisdom as a cognitive and motivational metaheuristic (pragmatic) that organizes and orchestrates knowledge toward human excellence in mind and virtue, both individually and collectively. PMID- 11392857 TI - States of excellence. AB - Research from the individual-differences tradition pertinent to the optimal development of exceptional talent is reviewed, using the theory of work adjustment (TWA) to organize fundings. The authors show how TWA concepts and psychometric methods, when used together, can facilitate positive development among talented youth by aligning learning opportunities with salient aspects of each student's individuality. Longitudinal research and more general theoretical models of (adult) academic and intellectual development support this approach. This analysis also uncovers common threads running through several positive psychological concepts (e.g., effectance motivation, flow, and peak experiences). The authors conclude by underscoring some important ideals from counseling psychology for fostering intellectual development and psychological well-being. These include conducting a multifaceted assessment, focusing on strength, helping people make choices, and providing a developmental context for bridging educational and industrial psychology to facilitate positive psychological growth throughout the life span. PMID- 11392858 TI - The evolution of happiness. AB - An evolutionary perspective offers novel insights into some major obstacles to achieving happiness. Impediments include large discrepancies between modern and ancestral environments, the existence of evolved mechanisms "designed" to produce subjective distress, and the fact that evolution by selection has produced competitive mechanisms that function to benefit one person at the expense of others. On the positive side, people also possess evolved mechanisms that produce deep sources of happiness: those for mating bonds, deep friendship, close kinship, and cooperative coalitions. Understanding these psychological mechanisms -the selective processes that designed them, their evolved functions, and the contexts governing their activation--offers the best hope for holding some evolved mechanisms in check and selectively activating others to produce an overall increment in human happiness. PMID- 11392859 TI - Creativity. Cognitive, personal, developmental, and social aspects. AB - Although many psychologists have expressed an interest in the phenomenon of creativity, psychological research on this topic did not rapidly expand until after J. P. Guilford claimed, in his 1950 APA presidential address, that this topic deserved far more attention than it was then receiving. This article reviews the progress psychologists have made in understanding creativity since Guilford's call to arms. Research progress has taken place on 4 fronts: the cognitive processes involved in the creative act, the distinctive characteristics of the creative person, the development and manifestation of creativity across the individual life span, and the social environments most strongly associated with creative activity. Although some important questions remain unanswered, psychologists now know more than ever before about how individuals achieve this special and significant form of optimal human functioning. PMID- 11392860 TI - The origins and ends of giftedness. AB - Five issues about giftedness are discussed. First, the origins of giftedness are explored. The view that giftedness is entirely a product of training is critiqued. There is indirect evidence for atypical brain organization and innate talent in gifted children: Many gifted children and savants have enhanced right hemisphere development, language-related difficulties, and autoimmune disorders. Second, the intense motivation of gifted children is discussed. Third, it is argued that gifted children have social and emotional difficulties that set them apart. Fourth, evidence for the often uneven cognitive profiles of such children is presented. Finally, the relationship between childhood giftedness and "domain" creativity in adulthood is discussed. Few gifted children go on to become adult creators because the skills and personality factors required to be a creator are very different from those typical of even the most highly gifted children. PMID- 11392861 TI - Toward a psychology of positive youth development. AB - This article analyzes the development of initiative as an exemplar of one of many learning experiences that should be studied as part of positive youth development. The capacity for initiative is essential for adults in our society and will become more important in the 21st century, yet adolescents have few opportunities to learn it. Their typical experiences during schoolwork and unstructured leisure do not reflect conditions for learning initiative. The context best suited to the development of initiative appears to be that of structured voluntary activities, such as sports, arts, and participation in organizations, in which youths experience the rare combination of intrinsic motivation in combination with deep attention. An incomplete body of outcome research suggests that such activities are associated with positive development, but the developmental processes involved are only beginning to be understood. One promising approach has recorded language use and has found that adolescents participating in effective organizations acquire a new operating language that appears to correspond to the development of initiative. PMID- 11392862 TI - Individual development in a bio-cultural perspective. AB - Biological and cultural inheritance deeply influence daily human behavior. However, individuals actively interact with bio-cultural information. Throughout their lives, they preferentially cultivate a limited subset of activities, values, and personal interests. This process, defined as psychological selection, is strictly related to the quality of subjective experience. Specifically, cross cultural studies have highlighted the central role played by optimal experience or flow, the most positive and complex daily experience reported by the participants. It is characterized by high involvement, deep concentration, intrinsic motivation, and the perception of high challenges matched by adequate personal skills. The associated activities represent the basic units of psychological selection. Flow can therefore influence the selective transmission of bio-cultural information and the process of bio-cultural evolution. PMID- 11392863 TI - Subjective well-being. The science of happiness and a proposal for a national index. AB - One area of positive psychology analyzes subjective well-being (SWB), people's cognitive and affective evaluations of their lives. Progress has been made in understanding the components of SWB, the importance of adaptation and goals to feelings of well-being, the temperament underpinnings of SWB, and the cultural influences on well-being. Representative selection of respondents, naturalistic experience sampling measures, and other methodological refinements are now used to study SWB and could be used to produce national indicators of happiness. PMID- 11392864 TI - The future of optimism. AB - Recent theoretical discussions of optimism as an inherent aspect of human nature converge with empirical investigations of optimism as an individual difference to show that optimism can be a highly beneficial psychological characteristic linked to good mood, perseverance, achievement, and physical health. Questions remain about optimism as a research topic and more generally as a societal value. Is the meaning of optimism richer than its current conceptualization in cognitive terms? Are optimism and pessimism mutually exclusive? What is the relationship between optimism and reality, and what are the costs of optimistic beliefs that prove to be wrong? How can optimism be cultivated? How does optimism play itself out across different cultures? Optimism promises to be one of the important topics of interest to positive social science, as long as it is approached in an even handed way. PMID- 11392865 TI - Positive psychology. An introduction. AB - A science of positive subjective experience, positive individual traits, and positive institutions promises to improve quality of life and prevent the pathologies that arise when life is barren and meaningless. The exclusive focus on pathology that has dominated so much of our discipline results in a model of the human being lacking the positive features that make life worth living. Hope, wisdom, creativity, future mindedness, courage, spirituality, responsibility, and perseverance are ignored or explained as transformations of more authentic negative impulses. The 15 articles in this millennial issue of the American Psychologist discuss such issues as what enables happiness, the effects of autonomy and self-regulation, how optimism and hope affect health, what constitutes wisdom, and how talent and creativity come to fruition. The authors outline a framework for a science of positive psychology, point to gaps in our knowledge, and predict that the next century will see a science and profession that will come to understand and build the factors that allow individuals, communities, and societies to flourish. PMID- 11392866 TI - The funds, friends, and faith of happy people. AB - New studies are revealing predictors of subjective well-being, often assessed as self-reported happiness and life satisfaction. Worldwide, most people report being at least moderately happy, regardless of age and gender. As part of their scientific pursuit of happiness, researchers have examined possible associations between happiness and (a) economic growth and personal income, (b) close relationships, and (c) religious faith. PMID- 11392867 TI - Self-determination theory and the facilitation of intrinsic motivation, social development, and well-being. AB - Human beings can be proactive and engaged or, alternatively, passive and alienated, largely as a function of the social conditions in which they develop and function. Accordingly, research guided by self-determination theory has focused on the social-contextual conditions that facilitate versus forestall the natural processes of self-motivation and healthy psychological development. Specifically, factors have been examined that enhance versus undermine intrinsic motivation, self-regulation, and well-being. The findings have led to the postulate of three innate psychological needs--competence, autonomy, and relatedness--which when satisfied yield enhanced self-motivation and mental health and when thwarted lead to diminished motivation and well-being. Also considered is the significance of these psychological needs and processes within domains such as health care, education, work, sport, religion, and psychotherapy. PMID- 11392868 TI - Self-determination. The tyranny of freedom. AB - Americans now live in a time and a place in which freedom and autonomy are valued above all else and in which expanded opportunities for self-determination are regarded as a sign of the psychological well-being of individuals and the moral well-being of the culture. This article argues that freedom, autonomy, and self determination can become excessive, and that when that happens, freedom can be experienced as a kind of tyranny. The article further argues that unduly influenced by the ideology of economics and rational-choice theory, modern American society has created an excess of freedom, with resulting increases in people's dissatisfaction with their lives and in clinical depression. One significant task for a future psychology of optimal functioning is to deemphasize individual freedom and to determine which cultural constraints are necessary for people to live meaningful and satisfying lives. PMID- 11392869 TI - Adaptive mental mechanisms. Their role in a positive psychology. AB - Psychology needs a metric for positive mental health that would be analogous to the IQ tests that measure above-average intelligence. The Defensive Function Scale of the DSM-IV offers a possible metric. In the present article the author links the transformational qualities of defenses at the mature end of the Defensive Function Scale--altruism, suppression, humor, anticipation, and sublimation--to positive psychology. First, the methodological problems involved in the reliable assessment of defenses are acknowledged. Next, the use of prospective longitudinal study to overcome such difficulties and to provide more reliable definition and measurement of defenses is outlined. Evidence is also offered that, unlike many psychological measures, the maturity of defenses is quite independent of social class, education, and IQ. Last, evidence is offered to illustrate the validity of mature defenses and their contribution to positive psychology. PMID- 11392870 TI - Psychological resources, positive illusions, and health. AB - Psychological beliefs such as optimism, personal control, and a sense of meaning are known to be protective of mental health. Are they protective of physical health as well? The authors present a program of research that has tested the implications of cognitive adaptation theory and research on positive illusions for the relation of positive beliefs to disease progression among men infected with HIV. The investigations have revealed that even unrealistically optimistic beliefs about the future may be health protective. The ability to find meaning in the experience is also associated with a less rapid course of illness. Taken together, the research suggests that psychological beliefs such as meaning, control, and optimism act as resources, which may not only preserve mental health in the context of traumatic or life-threatening events but be protective of physical health as well. PMID- 11392871 TI - The case for getting your MBA. PMID- 11392873 TI - The roadmap to significance (Part 2). PMID- 11392872 TI - The horror of suicide and child killing. PMID- 11392874 TI - Recommendations for processing and reporting of lymph node specimens submitted for evaluation of metastatic disease. PMID- 11392875 TI - DNA ploidy and cyclin D1 expression in basal cell carcinoma of the head and neck. AB - Basal cell carcinomas (BCCs) may be subdivided into primary with a favorable biologic course (BCC1) and recurrent and/or metastatic (BCC2). No clear association between primary tumor location, histologic subtype, or other clinicopathologic variables and predisposition for BCC2 has been found. Histopathologic criteria are limited for prognostication. To identify prognostic factors useful for planning therapy, we studied cyclin D1 immunohistochemical expression, DNA ploidy, and epiluminescence light microscopic (ELM) patterns in 60 cases of BCC (30 BCC1 and 30 BCC2) in the head and neck region, half of which were hyperpigmented. Cyclin D1 was absent in 27 cases, expressed at low level in 4 cases, and overexpressed in 30 cases. Seven BCCs were euploid, 28 exhibited a mixed cellular population, and 25 were aneuploid. Among aneuploid tumors, hypodiploidy was found in 12. Among the 30 pigmented carcinomas, only 15 showed a typical ELM pattern. No association between pigmentation and more aggressive biologic behavior of BCC was found. These results and follow-up data seem to indicate that an unfavorable outcome can be predicted by hyperexpression of cyclin D1, aneuploidy, and an atypical ELM pattern for pigmented cases. A definite hypodiploid peak was associated with worse prognosis. The analysis of cyclin D1 expression and DNA ploidy may help identify BCC with an aggressive phenotype and a poor clinical outcome. PMID- 11392876 TI - Quantitative evaluation of HER-2/neu status in breast cancer by fluorescence in situ hybridization and by immunohistochemistry with image analysis. AB - We correlated quantitative results obtained in 40 invasive breast cancer cases for HER-2 gene amplification by fluorescence in situ hybridization with protein expression by immunohistochemical studies with computer-assisted image analysis. Fluorescence in situ hybridization (FISH) results were quantified as the mean number of fluorescent signals per nucleus, and immunohistochemical slides were read by semiquantitatively assessing membranous immunostaining intensity in tumor cells vs nonneoplastic breast tissue or quantitatively evaluated by image analysis. We found high correlation between immunohistochemical results by semiquantitative scoring and by image analysis. FISH results correlated with immunohistochemical results moderately when the staining intensity of only tumor cells was assessed and significantly better when the difference in staining intensity between tumor cells and nonneoplastic breast tissue was assessed. The correlation with FISH results was further improved when immunohistochemical study was combined with heat-induced epitope retrieval (HIER). Although FISH and immunohistochemical studies assess different aspects of the HER-2/neu gene (amplification vs overexpression), we found good correlation between the diagnostic techniques. The correlation was best when immunohistochemical studies were combined with HIER and assessed as the difference between tumor cells and nonneoplastic breast tissue. PMID- 11392877 TI - Microsatellite instability is infrequent in medullary breast cancer. AB - Microsatellite instability (MSI), characterized by contraction or expansion in microsatellite length or short tandem repeats compared with germline lengths, is found in 85% to 90% of colon cancer arising in hereditary nonpolyposis colorectal cancer families. These cancers commonly have characteristic histologic appearances, including medullary features with intense lymphoid infiltrates. In pancreatic cancer, a rare medullary histologic subtype more often demonstrates MSI than the more common adenocarcinoma subtype. We hypothesized that the medullary histologic pattern might correlate with MSI in additional tumor types and analyzed 8 cases of typical and atypical medullary carcinoma of the breast. Tumor and normal DNA was extracted from paraffinized tissue blocks of tumor and histologically uninvolved axillary lymph nodes, respectively. We analyzed the tumors for instability in 5 primary (BAT25, BAT26, D17S250, D5S346, D2S123) and 3 alternative (BAT40, D18S55, D18S58) microsatellites recommended at the National Cancer Institute--sponsored conference for diagnosis of MSI in colorectal cancer. All 8 tumors were microsatellite stable at the 8 loci, suggesting that MSI is not commonly associated with medullary or atypical medullary breast carcinoma, in contrast with the reported association with medullary tumors of the colon and pancreas. PMID- 11392878 TI - Chromosomal imbalances in gastric cancer. Correlation with histologic subtypes and tumor progression. AB - DNA copy number changes were analyzed by comparative genomic hybridization (CGH) in 38 gastric carcinomas and correlated with tumor histologic type and progression. Gains of copy numbers were observed in all tumors, affecting all chromosomes except chromosome 16. The average number of copy number gains was 7 (range, 1-13), most frequently located on chromosomes 11, 12, 15, 17, and 20 in 45% to 97% of tumors. High-level amplifications were found on chromosomes 12, 15, 17, and 20; the latter was affected most frequently (66%). Loss of DNA copy numbers was detected in 14 tumors affecting 7 chromosomes. No statistically significant differences in the frequency and pattern of chromosomal imbalances were observed in tumor histologic type (Lauren classification) and grade of differentiation, as well as the prognostic parameters depth of invasion (pT) and lymph node involvement (pN). Our results indicate that in gastric cancer there is no specific recurrent pattern of DNA aberrations to be correlated with tumor histologic type or stage. However, CGH analysis could reveal new, recurrent genetic changes in gastric cancer affecting chromosomes sites that harbor genes known to participate in tumorigenesis and progression of several human malignant neoplasms. PMID- 11392879 TI - Frequent c-myc amplification in high-grade dysplasia and adenocarcinoma in Barrett esophagus. AB - Barrett esophagus (BE) is a condition in which the normal squamous epithelium of the esophagus is replaced by a metaplastic columnar epithelium. BE is a premalignant lesion that represents the initial step in a metaplasia-dysplasia carcinoma sequence. In the present study, amplification of the proto-oncogene c myc was determined by means of differential polymerase chain reaction analysis of metaplastic specialized epithelium, low-grade dysplasia, high-grade dysplasia, and invasive adenocarcinoma obtained by microscopic dissection of 43 esophagectomy specimens. Amplification of c-myc was found in none of 29 specialized epithelial specimens, none of 23 low-grade dysplasia specimens, 6 of 24 high-grade dysplasia specimens, and 17 of 39 adenocarcinoma specimens. Our data indicate that amplification of c-myc is a late event in the metaplasia dysplasia-carcinoma sequence in BE. Furthermore, determination of c-myc amplification may help identify high-risk patients who would benefit from intensified endoscopic surveillance or from immediate treatment. PMID- 11392881 TI - Endocervical status is not predictive of the incidence of cervical cancer in the years after negative smears. AB - The clinical relevance of the lack of endocervical cells was never well established in a longitudinal study with histologically proven cervical cancer as an end point. From the Dutch Network and National Database for Pathology, results for all negative smears obtained in 1990 and 1991 in the Netherlands were retrieved, as were data for all cytologic and histologic examinations performed after the negative smears before April 1998. There were no significant differences between the proportion of preinvasive lesions (cervical intraepithelial neoplasia 1, 2, and 3) detected after negative smears without endocervical cells compared with negative smears with endocervical cells. The proportion of women in whom invasive cancer developed was the same in both groups. These data suggest there is no reason to advise women with negative smears without endocervical cells to undergo an additional smear. PMID- 11392880 TI - Lymphoepithelioma-like carcinoma of the lung with a better prognosis. A clinicopathologic study of 32 cases. AB - The purpose of our study was to clarify the prognosis of lymphoepithelioma-like carcinoma (LELC) of the lung, which is rare. We analyzed the clinicopathologic features of 32 cases of pulmonary LELC and compared the cases with 84 cases of pulmonary non-LELC with available long-term follow-up information. The results show that LELC of the lung as a distinct entity has a better prognosis than non LELC. We found a significant difference in the survival rates between patients with LELC and patients with non-LELC in stage II and stages III and IV, respectively. Tumor recurrence and necrosis (5% or more of tumor) are associated with a poor prognosis. It seems that the histologic typing (Regaud type and Schmincke type) of pulmonary LELC is of no clinical value. PMID- 11392882 TI - Histologic classification of tumor-infiltrating lymphocytes in primary cutaneous malignant melanoma. A study of interobserver agreement. AB - The density and distribution of lymphocytes infiltrating the vertical growth phase of primary cutaneous melanomas has been suggested by several studies to be of prognostic significance. However, few pathologists comment on tumor infiltrating lymphocytes (TILs), and there is the perception that the assessment of TILs is subject to great interobserver variability. We studied interobserver agreement on the categorization of TILs; 20 cases of primary cutaneous malignant melanoma with a vertical growth phase component were circulated among 3 pathologists and 3 dermatologists. For each case, TILs were classified as brisk, nonbrisk, or absent according to Clark. Only 1 pathologist (a dermatopathologist) was familiar with the classification of TILs. Observers were given written guidelines and a brief tutorial before their examination of the slides. Our results show that with little instruction, overall agreement among observers was good (kappa values, 0.6 or more), especially among pathologists (kappa values, > 0.7). Three observers had excellent agreement among each other (kappa values, > 0.75). These findings suggest that the categorization of TILs can be easily taught and can be applied with an acceptable level of reproducibility in routine diagnostic practice. PMID- 11392883 TI - Decreased CD10 expression in grade III and in interfollicular infiltrates of follicular lymphomas. AB - CD10 expression in various grades and interfollicular infiltrates of follicular lymphoma (FL) has not been well documented. Immunohistochemical staining for CD10 (clone 56C6) was performed on paraffin-embedded tissue from 26 cases of classic FL. Negative or weak expression of CD10 was more frequent in grade III (5/6 [83%]) than in grade I FLs (3/15 [20%]). CD10+ interfollicular infiltrates were present in 16 cases. Six (38%) of 16 cases showed that CD10 expression was strong or moderate in follicular areas but weak or negative in interfollicular infiltrates. Our results suggest that CD10 expression is frequently weak to negative in grade III and in interfollicular infiltrates of FLs. Therefore, lack of CD10 expression on small specimens, such as from needle core biopsy or fine needle aspiration, does not preclude the possibility of a diagnosis of FL. Furthermore, lack of CD10 expression in diffuse large B-cell lymphoma does not exclude the possibility that the neoplastic lymphocytes are of follicle center cell origin. PMID- 11392884 TI - Precursor B-cell lymphoblastic lymphoma. A study of nine cases lacking blood and bone marrow involvement and review of the literature. AB - We describe 9 cases of precursor B-cell lymphoblastic lymphoma (LYL) without evidence of marrow or blood involvement. Four patients had superficial nodal disease, 2 cutaneous involvement, and 1 each ovarian, retroperitoneal, or tonsillar primary tumor. Six patients had limited disease; 3 patients were stage III. Immunophenotyping revealed a terminal deoxynucleotidyl transferase (TdT) positive, immature B-cell population with variable expression of CD10, CD20, and CD45. All patients are in complete clinical remission (median follow-up, 14 months). A literature review yielded 105 patients with a diagnosis of precursor B cell LYL based on less than 25% marrow involvement. Of these, 64% were younger than 18 years. Skin, lymph nodes, and bone were the most common sites of disease. Mediastinal involvement was uncommon. TdT, CD19, CD79a, CD10, and HLA-DR were the most frequently expressed antigens, while CD45 and CD20 were expressed in only two thirds of the cases. Cytogenetic analysis showed additional 21q material as a recurring karyotypic abnormality. At a median follow-up of 26 months, 74% of patients were alive; the median survival was 19 months for patients dying of disease. Comparison with precursor B-cell acute lymphoblastic leukemia showed several overlapping features, although distinct differences were identified. PMID- 11392885 TI - Diagnostic criteria for minimally differentiated acute myeloid leukemia (AML-M0). Evaluation and a proposal. AB - We studied immunophenotypic features of 30 cases of minimally differentiated acute myeloid leukemia (AML-M0) using multiparameter flow cytometry and immunohistochemistry and evaluated the immunophenotypic features of previously reported cases to facilitate correct identification of myeloid lineage. All but 1 of our 30 cases expressed CD13 and/or CD33; 2 expressed CD19; 1 expressed CD10; none expressed both CD10 and CD19. Eleven of 30 cases expressed T-cell-associated antigens. All but 2 cases expressed CD34 and/or HLA-DR. Twelve of 27 cases expressed terminal deoxynucleotidyl transferase. Myeloperoxidase (MPO) expression was seen in 22 of 22 cases by immunohistochemistry and 1 of 4 by flow cytometry. None of 27 cases expressed cyCD3 and cyCD79a. We propose following modified criteria for AML-M0: (1) standard criteria for acute leukemia; (2) undetectable or less than 3% MPO or Sudan black B staining in blasts; (3) lack of expression of lymphoid-specific antigens, cyCD3 for T lineage and cyCD79 and cyCD22 for B lineage; and (4) positivity for any of the myelomonocytic lineage antigens known not to be expressed on normal T or B lymphocytes or positivity for MPO as detected by ultrastructural cytochemistry, immunohistochemistry, or flow cytometry. PMID- 11392886 TI - Absence of CD26 expression is a useful marker for diagnosis of T-cell lymphoma in peripheral blood. AB - We report flow cytometric characterization of surface CD26 expression in 271 peripheral blood samples from 154 patients evaluated for the presence of a T-cell lymphoproliferative disorder, primarily mycosis fungoides/Sezary syndrome (MF/SS). The presence of morphologically identifiable tumor cells on peripheral blood smears was the criterion for lymphomatous involvement. In 66 of 69 samples from 28 patients, we identified an abnormal CD26-/dim T-cell population that was distinct from the variable CD26 expression seen in normal peripheral blood T cells. This population was CD26- in 23 patients and weakly CD26+ in 5 patients. CD7 was more variably expressed in MF/SS tumor cells, allowing recognition of a distinct, quantifiable abnormal T-cell population in only 34 of 69 involved samples. An increased CD4/CD8 ratio and lower surface expression of CD4 in tumor cells also helped separate the CD26-/dim atypical population for quantification. In 35 blood samples from other types of T-cell tumors, tumor cells in 10 of 11 morphologically involved cases showed absent/dim CD26. Although capable of detecting abnormalities in most cases of MF/SS, CD7 expression does not provide as clear a separation of the neoplastic population and can be replaced by CD26 staining in routine peripheral blood flow cytometric screening of MF/SS patients. PMID- 11392887 TI - Reduction of the platelet review rate using the two-dimensional platelet method. AB - Accurate platelet enumeration is critical for optimal treatment of patients with platelet and bleeding disorders, leukemias, and other neoplasias. The majority of automated hematology analyzers count platelets by size differentiation alone, which may result in falsely elevated platelet counts for samples containing interfering particles such as RBC fragments, microcytes, and cell debris. Most analyzers flag questionable platelet counts, necessitating review of results with confirmation by an alternative method, thus increasing the cost of performing platelet counts and delaying results. We studied the effect of a new platelet analysis method, based on measurement of size and refractive index, on the laboratory review rate for platelet counting. We demonstrated that this method yields higher accuracy for platelet counts in samples with interferences, especially for platelet counts less than 50 x 10(3)/microL (< 50 x 10(9)/L). As a result of the 2-dimensional analysis, the review rate for platelet counts was reduced by 65% in our institution, resulting in substantial savings. PMID- 11392888 TI - DNA amplification for the diagnosis of cat-scratch disease in small-quantity clinical specimens. AB - Diagnosis of cat-scratch disease (CSD) by polymerase chain reaction (PCR) of lymph node fineneedle aspiration (FNA) and primary lesion specimens can be difficult owing to the minute amount of available material. A PCR assay specifically suited to test these specimens was developed. First, small-quantity (10 microL) samples were prepared from 17 CSD-positive and 16 CSD-negative specimens, and DNA extraction and amplification from these samples were compared using 3 methods. Sensitivity and specificity of PCR were 100% using material collected on glass microscope slides and by using Qiagen (Hilden, Germany) columns for DNA extraction. Then, this method was used to test 11 archival glass microscope slides of FNA (7 malignant neoplasms, 4 undiagnosed lymphadenitis) and 2 primary lesion specimens. Two of the 4 lymphadenitis samples and the 2 primary lesion specimens were PCR positive. The technique presented could facilitate CSD diagnosis from a wider range of clinical samples. PMID- 11392889 TI - Ewing sarcoma vs lymphoblastic leukemia. PMID- 11392890 TI - Mortality risks, costs, and decision making in transfusion medicine. PMID- 11392891 TI - Promotion and tenure policies. PMID- 11392892 TI - [Comparison of quantitative coronary arteriography using cinefilm and digital images]. AB - OBJECTIVES: Quantitative coronary arteriography using both analog data (cinefilm) and digital data was evaluated with a stenotic vessel phantom and clinical data. METHODS: A stenotic vessel phantom using contrast medium with diameters of 2, 3, 4 and 6 mm was imaged using an X-ray image intensifier. Quantitative coronary arteriography was performed to measure reference and stenosis diameters in the phantom and under clinical conditions to measure coronary artery diameters using cinefilm and digital images. RESULTS: Both cinefilm and digital images were distorted at the peripheral image area, so errors were caused when the reference and stenosis were not in the same area. Measurements of diameters in the horizontal direction were larger than in the vertical and diagonal directions (p < 0.05). Reference diameters measured on digital images were larger than those on analog images (p < 0.0001). The difference in the real and measured reference diameter (2 and 3 mm) was less than 1% on digital images. The mean error in measurement of stenotic diameter measured on cinefilm images was 0.4-6.3% smaller for 0.5-4.5 mm diameter, but on digital images was 13% larger for 1 mm diameter, 18% larger for 0.75 mm diameter and 28% larger for 0.5 mm diameter. Under clinical conditions, the reference vessel diameter measured at the stenosis and the percentage stenosis were well correlated between the analog and digital images. CONCLUSIONS: Reference diameters of stenotic vessels are almost the same on cinefilm and digital images, but stenotic diameters under 1 mm are larger than actual size on digital images and on cinefilm images. PMID- 11392893 TI - [Analysis of factors influencing left ventricular mass and diastolic function in normotensive men]. AB - OBJECTIVES: Increase in left ventricular weight is an important risk factor for the incidence of cardiovascular diseases, and reduction in diastolic function of the left ventricle is an early marker for cardiac dysfunction. Factors related to the left ventricular mass and diastolic function were analyzed in middle-aged normotensive men. METHODS: The subjects were 126 normotensive men aged 49 +/- 1 years who were hospitalized for health-checkup. In addition to physical examination and routine laboratory tests, echocardiography including the pulse Doppler method was performed and urinary electrolyte excretions, plasma angiotensin II, plasma noradrenaline and the angiotensin converting enzyme genotype were examined. RESULTS: Left ventricular mass index was positively correlated with mean blood pressure (r = 0.249, p < 0.006) and body mass index (r = 0.279, p < 0.002). With regard to the index of left ventricular diastolic dysfunction, the late to early peak transmitral flow velocity ratio (A/E) was positively correlated with age (r = 0.465, p < 0.001) and urinary sodium excretion (r = 0.240, p < 0.007). Neither left ventricular mass index or A/E was affected by the angiotensin converting enzyme genotype and was not significantly correlated with plasma angiotensin II or noradrenaline. CONCLUSIONS: Increase in left ventricular mass is influenced by blood pressure and obesity, whereas reduction in left ventricular diastolic function is affected by greater age and salt intake. PMID- 11392894 TI - [Accuracy and usefulness of ultraportable hand-carried echocardiography system]. AB - OBJECTIVES: The diagnostic accuracy and usefulness of an ultraportable hand carried echocardiography system were investigated for assessing ventricular systolic function and severity of mitral valvular regurgitation. METHODS: The study population consisted of 77 consecutive patients (47 men, 30 women, mean age 63 +/- 15 years). Left ventricular end-diastolic dimension, left ventricular end systolic dimension and left ventricular ejection fraction were measured using the hand-carried echo system and the data were compared with measurements by the conventional echocardiography system using simple linear regression analysis. Left ventricular wall motion was compared between the systems using a 16-segment model recommended by the American Society of Echocardiography. Severity of mitral regurgitation was assessed by the distance of the regurgitant signal in the left atrium. RESULTS: Left ventricular end-diastolic dimension, left ventricular end systolic dimension and left ventricular ejection fraction showed good correlations between hand-carried and conventional echo systems (r = 0.94, 0.91 and 0.81, respectively; each p < 0.0001). The accuracy for assessing left ventricular wall motion was 94% (449 of 480 segments). The echo systems also showed the same degree of diagnostic accuracy for severity of mitral regurgitation. CONCLUSIONS: The hand-carried echo system provides accurate assessment of left ventricular function and mitral regurgitation simular to conventional echo machines. PMID- 11392895 TI - [Early outcome with the Alfieri mitral valve repair]. AB - OBJECTIVES: Mitral regurgitation in cases of prolapse of the anterior leaflet, posterior leaflet with calcified annulus, or prolapse of both leaflets is thought to be difficult to repair. The Alfieri repair has been developed to address these conditions. METHODS: Seven patients (four men and three women, mean age 71 +/- 9 years) underwent the Alfieri repair for mitral regurgitation at Austin and Repatriation Medical Centre between January 1999 and December 1999. The mechanism of mitral regurgitation was prolapse of the posterior leaflet with calcified annulus in one patient, prolapse of the anterior leaflet in two, and prolapse of both leaflets in four. Mitral regurgitation before operation was severe in all patients. The Cosgrove ring was used in all patients. Four patients underwent combined operation, coronary artery bypass surgery in three and tricuspid annuloplasty in one. RESULTS: There was no hospital death. Two patients had postoperative complications, transient ischemic attack in one patient and rapid atrial fibrillation in one. The mean hospital stay was 11.3 +/- 8.7 days. Mitral regurgitation after operation was mild in five patients and trivial in two. Mean pressure gradient of the transmitral valve was 4.0 +/- 1.4 mmHg. CONCLUSIONS: The Alfieri mitral valve repair is a simple and satisfactory technique to repair mitral regurgitation in selected patients. Long-term follow-up is required to evaluate the durability of this technique. PMID- 11392896 TI - [Impact of high remnant-like lipoproteins on the function of peripheral arterial endothelium in patients with hyperlipidemia and normolipidemia: estimation using Doppler flow analysis]. AB - OBJECTIVES: Cholesterol levels of remnant-like lipoprotein (RLP-cholesterol: RLP C), which reflect remnant lipoproteins, are associated with coronary arterial endothelial functions and cardiovascular events. The influence of RLP-C on peripheral vascular endothelial functions in patients with hyperlipidemia and normolipidemia was evaluated based on the reactivity of the antecubital artery to acetylcholine (Ach) and isosorbide dinitrate (ISDN) using a Doppler guidewire. METHODS: Protocol 1: Five patients were selected, and the dose-response of the antecubital artery was evaluated by administering Ach (0.5, 5, and 50 micrograms/30 sec) or ISDN (0.025, 0.25, and 2.5 mg/30 sec). An index of vascular reactivity (brachial artery response: BR) was determined by dividing the maximal blood flow velocity after the administration of Ach or ISDN by resting blood flow velocity. Protocol 2: BR was evaluated in 48 patients after administering Ach 50 micrograms or ISDN 2.5 mg. Subsequently, these patients were divided into the following three groups based on early morning RLP-C levels: Group L (n = 11), RLP C < 2.0 mg/dl (minimal detectable level); Group M (n = 21), 2.0 mg/dl < or = RLP C < 5.0 mg/dl; and Group H (n = 16), 5.0 mg/dl < or = RLP-C. The factors that regulate BR-Ach 50 micrograms were also evaluated in 34 normolipidemic patients. RESULTS: Protocol 1: BR dose-dependently increased after the administration of Ach and ISDN. Protocol 2: BR-Ach 50 micrograms was significantly less in Group H (3.1 +/- 0.8) than in Groups M (3.8 +/- 0.9, p < 0.03) and L (4.2 +/- 0.9, p < 0.01). However, there were no significant differences in BR-ISDN 2.5 mg between the three groups. Univariate analysis in normolipidemic patients revealed that BR Ach 50 micrograms was correlated with age (r = -0.355, p < 0.05), RLP-C (r = 0.488, p < 0.01), low-density lipoprotein cholesterol (r = -0.382, p < 0.03), systolic blood pressure (r = -0.354, p < 0.05), and diastolic blood pressure (r = -0.406, p < 0.02). Multivariate analysis using these five factors as independent variables revealed that age, RLP-C, and low-density lipoprotein cholesterol regulated BR-Ach 50 micrograms. CONCLUSIONS: Peripheral vascular endothelial dysfunction may occur in patients with high levels of RLP-C. RLP-C is an independent lipid factor that regulates peripheral vascular endothelial functions even in normolipidemic patients. PMID- 11392897 TI - [Pulmonary hypertension associated with refractory hyperthyroidism: a case report]. AB - A 25-year-old woman was admitted to our hospital with goiter. The diagnosis was Grave's disease. Diagnostic transthoracic echocardiography revealed a hyperdynamic stage of the heart with right ventricular dilation. Doppler echocardiography showed mild to moderate tricuspid regurgitation and elevated systolic right ventricular pressure. Right heart catheterization revealed high cardiac output (9.49 l/min) and pulmonary hypertension (57 mmHg) with increased pulmonary vascular resistance and total pulmonary resistance. No intracardiac shunts were detected. Since neither thiomazole nor propylthiouracil was effective and both caused side effects, she underwent subtotal thyroidectomy. After the surgery, pulmonary hypertension improved and cardiac output normalized, but without normalization of pulmonary vascular resistance and total pulmonary resistance. Reversible pulmonary hypertension may occur in patients with hyperthyroidism. Increased pulmonary blood flow and sustained high pulmonary artery resistance were suspected as the causes of pulmonary hypertension. In addition, pulmonary endothelial dysfunction as a result of sustained increased pulmonary blood flow could be another cause of pulmonary hypertension. PMID- 11392898 TI - [A 61-year-old woman with vascular bruit and severely impaired left ventricular function. Takayasu's arteritis (acquired atypical coarctation) with hypertensive heart disease]. PMID- 11392899 TI - It's the culture, stupid: using the values priority index. PMID- 11392900 TI - Intermediate sanctions: IRS issues new temporary regulations. PMID- 11392902 TI - Quality through leadership. PMID- 11392901 TI - Innovation and change: can you afford not to? PMID- 11392903 TI - The health divide--how the sexes differ. PMID- 11392904 TI - Racial intermarriage pairings. AB - Most studies of racial intermarriage rely on the prevalence of intermarriage to measure the strength of group boundaries, without scrutinizing the nature of intermarriage pairings. Examination of intermarried couples' characteristics reveals (1) that intermarriages and endogamous marriages follow different patterns, and (2) that intermarriage pairings for some groups reflect a generalized racial status hierarchy. According to evidence from the 1990 U.S. Census PUMS, patterns in blacks' and Mexican Americans' marriages with whites suggest that a generalized racial status hierarchy disadvantages members of these minority groups. For marriages between Japanese Americans and whites, however, crossing the group boundary does not affect couples' characteristics. PMID- 11392905 TI - The salience of pan-national Hispanic and Asian identities in U.S. marriage markets. AB - In this paper I test whether "Hispanic" and "Asian" identities are salient in the U.S. marriage market. That is, I determine whether the different Asian and Hispanic national groups intermarry often enough to suggest that Asian and Hispanic pan-national identities are important. Analysis of census data from 1980 and 1990 suggests that both Hispanic and Asian pan-national identities are significant forces. Variations in the strength of pan-national Hispanic and Asian associations by region, education, and nativity are discussed. PMID- 11392906 TI - Black-white unions: West Indians and African Americans compared. AB - In this research we use 1990 PUMS data to compare the propensity for unions between African Americans and native whites with the propensity for unions between British West Indians and native whites. In addition, we distinguish women and men. Descriptive statistics indicate that West Indians, with the exception of men who arrived as adults, are more likely than African Americans to have white partners. After the introduction of controls for several correlates of intermarriage, however, West Indian men of any generation have lower exogamy rates than African American men, while exogamy rates are higher among West Indian women who arrived as children or who were born in the United States than among African American women. Thus we find no consistent evidence of greater exogamy for British West Indians than for African Americans. PMID- 11392907 TI - On the auspices of female migration from Mexico to the United States. AB - In this paper we examine the circumstances and determinants of female migration between Mexico and the United States. Using data from the Mexican Migration Project, we considered the relative timing of males' and females' moves northward. We then estimated logit and probit models to study the determinants of male and female out-migration; among women we also estimated a multinomial logit model to uncover differences in the process of migration for work versus not for work. We found that women almost always followed other family members, either the husband or a parent; only a tiny minority initiated migration independently. Although males also are quite likely to be introduced to migration by a parent, nearly half of all male migrants left for the United States before or without a wife or a parent. Estimates of the determinants of migration suggested that males move for employment, whereas wives generally are motivated by family reasons. Daughters, however, display a greater propensity to move for work, and the determinants of their work-related moves closely resemble those of sons and fathers. PMID- 11392908 TI - A cross-national comparison of the impact of family migration on women's employment status. AB - In this paper we consider the effects of family migration on women's employment status, using census microdata from Great Britain and the United States. We test a simple hypothesis that families tend to move long distances in favor of the male's career and that this can have a detrimental effect on women's employment status. Unlike many previous studies of this question, our work emphasizes the importance of identifying couples that have migrated together, rather than simply comparing long-distance (fe)male migrants with nonmigrant (fe)males individually. We demonstrate that women's employment status is harmed by family migration; the results we present are surprisingly consistent for Great Britain and the United States, despite differing economic situations and cultural norms regarding gender and migration. We also demonstrate that studies that fail to identify linked migrant couples are likely to underestimate the negative effects of family migration on women's employment status. PMID- 11392909 TI - Increasing longevity and Medicare expenditures. AB - Official Medicare projections forecast that the elderly population will be less healthy and more costly over the next century. This prediction stems from the use of age as an indicator of health status: increases in longevity are assumed to increase demand for health care as individuals survive to older and higher-use ages. In this paper I suggest an alternative approach, in which time until death replaces age as the demographic indicator of health status. Increases in longevity are assumed to postpone the higher Medicare use and costs associated with the final decade of life. I contrast the two approaches, using mortality forecasts consistent with recent projections from the U.S. Census Bureau and the Social Security Administration. The time-until-death method yields significantly lower-cost forecasts. The hypothetical cost savings from improved health care small, however, relative to the size of the Medicare solvency problem caused by population aging. PMID- 11392911 TI - Women's health and pregnancy outcomes: do services make a difference? AB - We use data from the Indonesia Family Life Survey to investigate the impact of a major expansion in access to midwifery services on health and pregnancy outcomes for women of reproductive age. Between 1990 and 1998 Indonesia trained some 50,000 midwives. Between 1993 and 1997 these midwives tended to be placed in relatively poor communities that were relatively distant from health centers. We show that additions of village midwives to communities between 1993 and 1997 are associated with a significant increase in body mass index in 1997 relative to 1993 for women of reproductive age, but not for men or for older women. The presence of a village midwife during pregnancy is also associated with increased birthweight. Both results are robust to the inclusion of community-level fixed effects, a strategy that addresses many of the concerns about biases because of nonrandom program placement. PMID- 11392910 TI - Inequality in life expectancy, functional status, and active life expectancy across selected black and white populations in the United States. AB - We calculated population-level estimates of mortality, functional health, and active life expectancy for black and white adults living in a diverse set of 23 local areas in 1990, and nationwide. At age 16, life expectancy and active life expectancy vary across the local populations by as much as 28 and 25 years respectively. The relationship between population infirmity and longevity also varies. Rural residents outlive urban residents, but their additional years are primarily inactive. Among urban residents, those in more affluent areas outlive those in high-poverty areas. For both whites and blacks, these gains represent increases in active years. For whites alone they also reflect reductions in years spent in poor health. PMID- 11392912 TI - Savings/credit group formation and change in contraception. AB - We examine the characteristics of women who chose to join a women's savings or credit group organized by Save the Children USA in a rural area of Bangladesh, and the impact of participation on contraceptive use. The data are taken from a panel survey conducted in 1993, shortly before the groups were formed, and in 1995 after interventions began. Our findings show that although demographic and socioeconomic characteristics have only a weak relationship to the decision to join a program, the treatment that a woman receives from her husband is associated with participation. We also find evidence that the credit program tends to attract women who are already using contraception. The analysis of program impact on the use of modern contraceptives reveals a positive effect of the credit program, after we adjust for this selectivity; we see no evidence of an effect of participation in a savings group. PMID- 11392913 TI - The influence of remarriage on the racial difference in mother-only families in 1910. AB - Historical demography documents that mother-only families were more common among African Americans than among Euro-Americans early in the twentieth century. We find direct evidence that African American males in both first and higher-order marriages were more likely to have (re)married previously married women and were more likely to have (re)married women with children. This racial difference in (re)marital partner choice reduced the racial difference in the prevalence of mother-only families such that, in the absence of such remarriage choices, the prevalence of mother-only families in the turn-of-the-century African American population would have been even higher than has been reported. Remarriage in this period countered the various demographic, economic, cultural, and social institutional forces that disproportionately destabilized African American marriages; it must be taken into account more fully by analysts concerned with racial differences in family structure. PMID- 11392914 TI - Availability of child care in the United States: a description and analysis of data sources. AB - Lack of high-quality, affordable, and accessible child care is an often-cited impediment to a manageable balance between work and family. Researchers, however, have been restricted by a scarcity of data on the availability of child care across all U.S. communities. In this paper we describe and evaluate several indicators of child care availability that have been released by the U.S. Census Bureau over the last 15 years. Using community- and individual-level analyses, we find that these data sources are useful for indicating child care availability within communities, even though they were collected for other purposes. Furthermore, our results generally suggest that the data on child care availability are equally valid across communities of different urbanicity and average income levels, although it appears that larger geographic areas more accurately capture the child care market of centers than that of family day care providers. Our analyses indicate that center child care is least available in nonmetropolitan, poor communities, and that family day care is most available in nonmetropolitan, mixed-income communities. We discuss the benefits and limitations of the data sources, and point to directions for future data developments and research. PMID- 11392915 TI - Couples' adjustment to breast disease during the first year following diagnosis. AB - The objectives were to determine (a) the extent to which psychosocial, demographic, and medical variables predict women's and husbands' adjustment to breast disease during the first year following diagnosis; (b) the degree of autocorrelation among and intercorrelation between partners' adjustment scores; (c) the extent to which baseline levels of adjustment predict adjustment 1 year later; and (d) the extent to which one partner's adjustment affects the other partner's adjustment. A stress-coping framework guided this study. The sample consisted of 131 couples, 58 couples received a cancer diagnosis and 73 received a benign diagnosis. Couples were interviewed at 1 week, 2 months, and 1 year postdiagnosis. Structural equation modeling was used to analyze the data. The strongest predictors of adjustment for women were severity of the illness and hopelessness and for husbands, their own baseline level of adjustment. Husbands' and wives' levels of adjustment at 1 year had a significant direct effect on each other's adjustment. PMID- 11392916 TI - The effects of stress, anxiety, and outdoor temperature on the frequency and severity of Raynaud's attacks: the Raynaud's Treatment Study. AB - It was expected that stress and anxiety would be related to Raynaud's phenomenon (RP) attack characteristics when mild outdoor temperatures produced partial or no digital vasoconstriction. Hypotheses were that in warmer temperature categories, compared to those below 40 degrees F, higher stress or anxiety would be associated with more frequent, severe, and painful attacks. The Raynaud's Treatment Study recruited 313 participants with primary RP. Outcomes were attack rate, severity, and pain. Predictors were average daily outdoor temperature, stress, anxiety, age, gender, and a stress-by-temperature or an anxiety-by temperature interaction. Outcomes were tested separately in multiple linear regression models. Stress and anxiety were tested in separate models. Stress was not a significant predictor of RP attack characteristics. Higher anxiety was related to more frequent attacks above 60 degrees F. It was also related to greater attack severity at all temperatures, and to greater pain above 60 degrees F and between 40 degrees and 49.9 degrees F. PMID- 11392917 TI - Differential fear of cardiopulmonary sensations in emergency room noncardiac chest pain patients. AB - Fear of bodily sensations has received extensive attention in relation to panic disorder, and more recently, other types of anxiety pathology and chronic pain problems. Extending this work, the present study examined fear of bodily sensations and its underlying dimensions in emergency room patients with Noncardiac Chest Pain (NCCP; n = 63). We posited a "differential specificity" hypothesis, expecting that specific cardiopulmonary fears would be more strongly associated with NCCP symptoms relative to other bodily fears. As hypothesized, participants reported cardiopulmonary sensations as significantly more fear provoking than numbness, dissociation, and gastrointestinal sensations. Additionally, regression analysis indicated that after accounting for theoretically relevant demographic variables and health status, cardiopulmonary fear was the best predictor of a composite index of cardiac complaints intensity, even after removing variance related to the absolute number of cardiac complaints. We discuss these findings in relation to the specific role for the fear of cardiopulmonary sensations in chest pain complaints, with implications for better understanding the underlying psychological processes involved in NCCP. PMID- 11392918 TI - Cognitive avoidance as a method of coping with a provocative smoking cue: the moderating effect of nicotine dependence. AB - Both nicotine dependence and coping are important determinants of smoking cessation, yet little is understood about mechanisms that link the two. This laboratory study investigated how nicotine dependence moderates execution of an avoidance coping strategy. High and low dependent smokers were exposed to a provocative smoking cue (in vivo) under two instructional sets: cognitive avoidance coping and no coping. Contrary to hypotheses, high dependent smokers reported greater increases in perceived self-efficacy to not smoke and also demonstrated greater facility in processing coping/nonsmoking-related information on a reaction time task, compared with low dependent smokers. These counterintuitive findings are discussed in terms of how nicotine dependence may affect the cognitive process of coping. PMID- 11392919 TI - Adolescent illegal drug use: the impact of personality, family, and environmental factors. AB - This study examined the relationship between the domains of environmental factors, family illegal drug use, parental child-rearing practices, maternal and adolescent personality attributes, and adolescent illegal drug use. A nonclinical sample of 2,837 Colombian youths and their mothers were interviewed about intrapersonal, interpersonal, and environmental factors in their lives. Results indicated that certain environmental factors (e.g., violence, drug availability, and machismo), family drug use, a distant parent-child relationship, and unconventional behavior are risk factors for adolescent illegal drug use. As hypothesized, results showed that the adverse effects of family illegal drug use on adolescent drug use can be buffered by protective parental child-rearing practices and environmental factors, leading to less adolescent illegal drug use. Prevention and treatment efforts should incorporate protective environmental, familial, and intrapersonal components in order to reduce adolescent illegal drug use. PMID- 11392921 TI - Why lesbian, gay, bisexual, and transgender public health? PMID- 11392920 TI - "I'm sorry to tell you ..." physicians' reports of breaking bad news. AB - In this investigation the authors assessed what physicians do when planning for and delivering bad news to patients. Seventy-three physicians responded to a series of statements about the behaviors, thoughts, and feelings they might have had while preparing for and delivering bad medically-related news. Data were also obtained about how well they thought the transaction had gone, how much stress they had experienced, and what they thought the experience was like from the patient's perspective. Physicians reported that these transactions were only moderately stressful, with 18.1% and 18.7% indicating that preparation stress or delivery stress, respectively, were above the midpoint on the scale. Slightly over 42% of the sample indicated that the stress they experienced lasted from several hours to three or more days. Thirty-six delivery-related statements were typical (with endorsement rates of at least 80% in a given direction) for at least one of the two recall groups. PMID- 11392922 TI - Uneasy promises: sexuality, health, and human rights. AB - Although attention to the links between health and human rights is growing globally, the full potential of a progressive human rights approach to health has not yet been explored, and it is even more faintly understood in the United States than in the rest of the world. At the same time, global claims for sexual rights, particularly for those identifying as gay, lesbian, transsexual, or bisexual, are increasingly being made as human rights claims. All of these approaches to rights advocacy risk limiting their own transformative impact unless advocates critique their own strategies. Paradoxically, using health as a way to bring attention to nonheteronormative sexualities can be both helpful and potentially dangerous, especially when coupled with human rights. Recognizing sexuality as a critical element of humanity, and establishing a fundamental human right to health, can play a role in broader social justice claims, but the tendency of both public health and human rights advocacy to "normalize" and regulate must be scrutinized and challenged. PMID- 11392924 TI - Enhancing transgender health care. AB - As awareness of transgender men and women grows among health care educators, researchers, policymakers, and clinicians of all types, the need to create more inclusive settings also grows. Greater sensitivity and relevant information and services are required in dealing with transgender men and women. These individuals need their identities to be recognized as authentic, they need better access to health care resources, and they need education and prevention material appropriate to their experience. In addition, a need exists for activities designed to enhance understanding of transgender health issues and to spur innovation. PMID- 11392923 TI - Ethics that exclude: the role of ethics committees in lesbian and gay health research in South Africa. AB - Prevailing state and institutional ideologies regarding race/ethnicity, gender, and sexuality help to shape, and are influenced by, research priorities. Research ethics committees perform a gatekeeper role in this process. In this commentary, we describe efforts to obtain approval from the ethics committee of a large medical institution for research into the treatment of homosexual persons by health professionals in the South African military during the apartheid era. The committee questioned the "scientific validity" of the study, viewing it as having a "political" rather than a "scientific" purpose. They objected to the framing of the research topic within a human rights discourse and appeared to be concerned that the research might lead to action against health professionals who committed human rights abuses against lesbians and gay men during apartheid. The process illustrates the ways in which heterosexism, and concerns to protect the practice of health professionals from scrutiny, may influence the decisions of ethics committees. Ethics that exclude research on lesbian and gay health cannot be in the public interest. Ethics committees must be challenged to examine the ways in which institutionalized ideologies influence their decision making. PMID- 11392925 TI - Undercounts and overstatements: will the IOM report on lesbian health improve research? AB - In January 1999, the Institute of Medicine (IOM) released a report on lesbian health research that fulfills 3 goals: it provides an extensive review of much of the research that has been done on the health of women who have sex with other women, it addresses the methodological and ethical issues inherent in conducting research on this population, and it suggests avenues for further research. This report will likely help lesbian health researchers gain funding, publish further research in medical journals, and receive support and validation from medical and research institutions. To ensure that such research is useful, benefits the lesbian community, and expands the understanding of lesbian health conditions, particular attention needs to be paid to the methods and definitions used and to the involvement of the lesbian community in designing, implementing, and analyzing the research itself. PMID- 11392926 TI - Sexual orientation data collection and progress toward Healthy People 2010. AB - Without scientifically obtained data and published reports, it is difficult to raise awareness and acquire adequate resources to address the health concerns of lesbian, gay, and bisexual Americans. The Department of Health and Human Services must recognize gaps in its information systems regarding sexual orientation data and take immediate steps to monitor and eliminate health disparities as delineated in Healthy People 2010. A paper supported by funding from the Office of the Assistant Secretary for Planning and Evaluation explores these concerns and suggests that the department (1) create work groups to examine the collection of sexual orientation data; (2) create a set of guiding principles to govern the process of selecting standard definitions and measures; (3) recognize that racial/ethnic, immigrant-status, age, socioeconomic, and geographic differences must be taken into account when standard measures of sexual orientation are selected; (4) select a minimum set of standard sexual orientation measures; and (5) develop a long-range strategic plan for the collection of sexual orientation data. PMID- 11392928 TI - Addressing lesbian, gay, bisexual, and transgender issues from the inside: one federal agency's approach. AB - The mission of the Substance Abuse and Mental Health Services Administration (SAMHSA) is to protect and serve underserved and vulnerable populations. Congress established SAMHSA under Public Law 102-321 on October 1, 1992, to strengthen the nation's health care capacity to provide prevention, diagnosis, and treatment services for substance abuse and mental illnesses. SAMHSA works in partnership with states, communities, and private organizations to address the needs of people with substance abuse and mental illnesses as well as the community risk factors that contribute to these illnesses. As part of its efforts to address the unique needs of special populations, SAMHSA has reached out to the lesbian, gay, bisexual, and transgender (LGBT) community. SAMHSA and its centers (Center for Substance Abuse Treatment, Center for Substance Abuse Prevention, and Center for Mental Health Services) have made a concerted effort, through both policy and programs, to develop services responsive to this community. PMID- 11392927 TI - Are we headed for a resurgence of the HIV epidemic among men who have sex with men? AB - HIV remains a critical health issue for men who have sex with men (MSM). In the United States, an estimated 365,000 to 535,000 MSM are living with HIV, and 42% of new HIV infections occur in this population. Recent data on sexually transmitted diseases and on sexual behavior indicate the potential for a resurgence in HIV infections among MSM. Outbreaks of gonorrhea and syphilis have been reported in a growing number of cities, and several studies have observed an increase in unprotected anal intercourse among MSM. These increases in HIV risk behavior may be attributed to several factors that have affected the sexual practices of MSM, including changes in beliefs regarding the severity of HIV disease. These emerging data have implications for surveillance and intervention research activities and indicate a need to reevaluate, refocus, and reinvigorate HIV prevention efforts for MSM. Our recommendations for addressing the HIV prevention needs of MSM include the need to consider HIV-related issues within the broader context of the physical, mental, and sexual health of MSM. PMID- 11392929 TI - The evolution of the Fenway Community Health model. AB - Fenway Community Health was founded by community activists in 1971 in the Fenway neighborhood of Boston, Mass, and within a decade had rapidly expanded its medical services for gay men in response to the AIDS epidemic. Increased expertise and cultural competence in lesbian, gay, bisexual, and transgender (LGBT) care led to expansion of medical services to address broader community concerns, ranging from substance use to parenting issues to domestic and homophobic violence, as well as specialized programs for lesbians, bisexuals, and transgendered individuals. Fenway began as a grassroots neighborhood clinic. In 1975, the center recorded 5000 patient care visits; in 2000, Fenway's clinical departments recorded 50,850 visits by 8361 individuals, including more than 1100 individuals receiving HIV-associated care. The center now has more than 170 staff people responsible for clinical programs, community education, research, administration, planning, and development. Over the past few years, Fenway's annual budget has exceeded $10 million. Fenway has established standards for improved cultural competence about LGBT health issues for other health providers and has developed programs to educate the general community about specific LGBT health concerns. This health center may provide a model of comprehensive LGBT health services that have a local impact. PMID- 11392930 TI - The GLBT Health Access Project: a state-funded effort to improve access to care. AB - The Gay, Lesbian, Bisexual, and Transgender (GLBT) Health Access Project is a unique public-private collaboration working to eliminate barriers to health care for the GLBT community, foster development of comprehensive, culturally appropriate health promotion policies and health care services for GLBT people and their families, and expand appropriate data collection on GLBT health. Funded by the Massachusetts Department of Public Health, the project developed community standards of practice for provision of quality health care services to GLBT clients. A health access training curriculum was developed and technical assistance was offered to health care providers implementing the standards, which cover personnel, clients' rights, intake and assessment, service delivery and planning, confidentiality, and community outreach and health promotion. Training participants (324 individuals from 89 agencies) reported positive though not statistically significant changes in attitude. PMID- 11392931 TI - Trans health crisis: for us it's life or death. PMID- 11392932 TI - Same-sex romantic attraction and experiences of violence in adolescence. AB - OBJECTIVES: Recent national attention to hate crimes committed against lesbian, gay, and bisexual youths has highlighted the need to understand this group's experiences of violence. Using nationally representative data, we examine the associations between romantic attraction and experiences of violence, as well as the risk of witnessing violence and perpetrating violence against others. METHODS: Data from the National Longitudinal Study of Adolescent Health were examined. Youths reporting same-sex and both-sex romantic attractions were compared with those reporting other-sex attractions. Survey logistic regression was used to control for sample design effects. RESULTS: Youths who report same sex or both-sex romantic attraction are more likely to experience extreme forms of violence than youths who report other-sex attraction. Youths reporting same sex and both-sex romantic attractions are also more likely to witness violence. The higher incidence of violence perpetrated by youths attracted to the same sex is explained by their experiences of violence. CONCLUSIONS: These findings provide strong evidence that youths reporting same-sex or both-sex romantic attraction are at greater risk for experiencing, witnessing, and perpetrating violence. PMID- 11392933 TI - The continuing HIV epidemic among men who have sex with men. AB - OBJECTIVES: This study characterized the AIDS epidemic among urban men who have sex with men (MSM). METHODS: A probability sample of MSM was obtained in 1997 (n = 2881; 18 years and older) from New York, Los Angeles, Chicago, and San Francisco, and HIV status was determined through self-report and biological measures. RESULTS: HIV prevalence was 17% (95% confidence interval = 15%, 19%) overall, with extremely high levels in African Americans (29%), MSM who used injection drugs (40%), "ultraheavy" noninjection drug users (32%), and less educated men (< high school, 37%). City-level HIV differences were non significant once these other factors were controlled for. In comparing the present findings with historical data based on public records and modeling, HIV prevalence appears to have declined as a result of high mortality (69%) and stable, but high, incidence rates (1%-2%). CONCLUSIONS: Although the findings suggest that HIV prevalence has declined significantly from the mid-1980s, current levels among urban MSM in the United States approximate those of sub Saharan countries (e.g., 14%-25%) and are extremely high in many population subsegments. Despite years of progress, the AIDS epidemic continues unabated among subsegments of the MSM community. PMID- 11392934 TI - HIV prevalence, risk behaviors, health care use, and mental health status of transgender persons: implications for public health intervention. AB - OBJECTIVES: This study described HIV prevalence, risk behaviors, health care use, and mental health status of male-to-female and female-to-male transgender persons and determined factors associated with HIV. METHODS: We recruited transgender persons through targeted sampling, respondent-driven sampling, and agency referrals; 392 male-to-female and 123 female-to-male transgender persons were interviewed and tested for HIV. RESULTS: HIV prevalence among male-to-female transgender persons was 35%. African American race (adjusted odds ratio [OR] = 5.81; 95% confidence interval [CI] = 2.82, 11.96), a history of injection drug use (OR = 2.69; 95% CI = 1.56, 4.62), multiple sex partners (adjusted OR = 2.64; 95% CI = 1.50, 4.62), and low education (adjusted OR = 2.08; 95% CI = 1.17, 3.68) were independently associated with HIV. Among female-to-male transgender persons, HIV prevalence (2%) and risk behaviors were much lower. Most male-to-female (78%) and female-to-male (83%) transgender persons had seen a medical provider in the past 6 months. Sixty-two percent of the male-to-female and 55% of the female-to male transgender persons were depressed; 32% of each population had attempted suicide. CONCLUSIONS: High HIV prevalence suggests an urgent need for risk reduction interventions for male-to-female transgender persons. Recent contact with medical providers was observed, suggesting that medical providers could provide an important link to needed prevention, health, and social services. PMID- 11392935 TI - Time-space sampling in minority communities: results with young Latino men who have sex with men. AB - OBJECTIVES: This study addressed methodological issues influencing the feasibility of time-space sampling in HIV prevention studies targeting hard-to reach populations of minority young men who have sex with men (MSM). METHODS: We conducted interviews with 400 men in 32 venues where young Latino MSM congregate in New York City. Response rates and demographic and sexual risk profiles are compared by venue type. RESULTS: More than 90% of the men approached were screened. Among eligible men, participation rates exceeded 82%. Participation was higher at special events and gay venues compared with nongay venues (P < .05). Young MSM in nongay venues were less likely to self-identify as gay (P < .01) or to report recent anal sex with a male (P < .10). Condom use did not vary by venue type but was lower with women than with men. If surveys had been limited to gay venues, about half of the young MSM surveyed in nongay venues would have been missed. CONCLUSIONS: Time-space sampling of a relatively "hidden" minority young MSM population can be successful across a range of venues. However, the benefits of greater outreach must be weighed against the costs incurred recruiting participants in nongay venues. PMID- 11392936 TI - The impact of homophobia, poverty, and racism on the mental health of gay and bisexual Latino men: findings from 3 US cities. AB - OBJECTIVES: This study assessed the relation between experiences of social discrimination (homophobia, racism, and financial hardship) and symptoms of psychologic distress (anxiety, depression, and suicidal ideation) among self identified gay and bisexual Latino men in the United States. METHODS: Data were collected from a probability sample of 912 men (self-identified as both Latino and nonheterosexual) recruited from the venues and public social spaces identified as both Latino and gay in the cities of Miami, Los Angeles, and New York. RESULTS: The study showed high prevalence rates of psychologic symptoms of distress in the population of gay Latino men during the 6 months before the interview, including suicidal ideation (17% prevalence), anxiety (44%), and depressed mood (80%). In both univariate and multivariate analyses, experiences of social discrimination were strong predictors of psychologic symptoms. CONCLUSIONS: The mental health difficulties experienced by many gay and bisexual Latino men in the United States are directly related to a social context of oppression that leads to social alienation, low self-esteem, and symptoms of psychologic distress. PMID- 11392937 TI - Risk of psychiatric disorders among individuals reporting same-sex sexual partners in the National Comorbidity Survey. AB - OBJECTIVES: This study examined the risk of psychiatric disorders among individuals with same-sex sexual partners. METHODS: Data are from the National Comorbidity Survey, a nationally representative household survey. Respondents were asked the number of women and men with whom they had sexual intercourse in the past 5 years. Psychiatric disorders according to Diagnostic and Statistical Manual of Mental Disorders, Revised Third Edition (DSM-III-R) criteria were assessed with a modified version of the Composite International Diagnostic Interview. RESULTS: A total of 2.1% of men and 1.5% of women reported 1 or more same-sex sexual partners in the past 5 years. These respondents had higher 12 month prevalences of anxiety, mood, and substance use disorders and of suicidal thoughts and plans than did respondents with opposite-sex partners only. Decomposition showed that the elevated same-sex 12-month prevalences were largely due to higher lifetime prevalences. Ages at onset and persistence of disorders did not differ between the same-sex and opposite-sex subsamples. CONCLUSIONS: Homosexual orientation, defined as having same-sex sexual partners, is associated with a general elevation of risk for anxiety, mood, and substance use disorders and for suicidal thoughts and plans. Further research is needed to replicate and explore the causal mechanisms underlying this association. PMID- 11392938 TI - Preventing sexual risk behaviors among gay, lesbian, and bisexual adolescents: the benefits of gay-sensitive HIV instruction in schools. AB - OBJECTIVES: This study compared sexual risk behaviors of gay, lesbian, and bisexual (GLB) and heterosexual adolescents and evaluated associations between gay-sensitive HIV instruction and risk behaviors of GLB youths. METHODS: A random sample of high school students and HIV education teachers completed surveys. Self reported risk behaviors of heterosexual and GLB adolescents were compared, with control for student and community demographic characteristics. Sexual risk behaviors of GLB youths in schools with and without gay-sensitive instruction were compared. RESULTS: GLB youths reported more substance use, high-risk sexual behaviors, suicidal thoughts or attempts, and personal safety issues than did heterosexual youths (P < .001). Among those who were sexually active, GLB youths reported more lifetime and recent sexual partners than did heterosexuals (P < .001), and more of them reported alcohol use before last sex (P < .01) and a history of pregnancy (P < .001). GLB youths in schools with gay-sensitive instruction reported fewer sexual partners, less recent sex, and less substance use before last sex than did GLB youths in other schools (P < .05). CONCLUSIONS: The findings document increased risk behaviors among GLB youths and demonstrate the potential benefits of providing gay-sensitive HIV instruction in schools. PMID- 11392939 TI - Papanicolaou test screening and prevalence of genital human papillomavirus among women who have sex with women. AB - OBJECTIVES: The purpose of this study was to examine frequency of and attitudes toward Papanicolaou (Pap) test screening in women who have sex with women (WSW) and to determine prevalence of genital human papillomavirus (HPV). METHODS: Women were eligible if they reported having engaged in sex with another woman in the preceding year Medical and sexual histories were obtained. Cervical specimens for Pap tests and cervical and vaginal specimens for HPV DNA testing were collected. RESULTS: HPV DNA was detected in 31 of 248 WSW (13%). Women who had never had sex with men were less likely to have undergone pelvic examinations and had fewer recent Pap tests. Reasons for not undergoing Pap tests included lack of insurance, previous adverse experiences, and belief that Pap tests were unnecessary. CONCLUSIONS: Despite the occurrence of genital HPV, WSW do not receive adequate Pap test screening. Pap test screening recommendations should not differ for WSW, regardless of sexual history with men. PMID- 11392940 TI - The Circuit Party Men's Health Survey: findings and implications for gay and bisexual men. AB - OBJECTIVES: This study examined characteristics of gay and bisexual men who attend circuit parties, frequency of and motivations for attending parties, drug use and sexual behavior during circuit party weekends, and use of risk reduction materials available at parties. METHODS: A cross-sectional survey was conducted among 295 gay and bisexual men from the San Francisco Bay Area who had attended a circuit party in the previous year. RESULTS: One fourth of the men reported a drug "overuse" incident in the previous year. Nearly all respondents reported use of drugs during circuit party weekends, including ecstasy (75%), ketamine (58%), crystal methamphetamine (36%), gamma hydroxybutyrate or gamma butyrolactone (25%), and Viagra (12%). Two thirds of the men reported having sex (oral or anal), 49% reported having anal sex, and 28% reported having unprotected anal sex during the 3-day period. An association was found between use of drugs and sexual risk behavior. Prevention materials were observed at party events by some men; however, relatively few men used the materials. Common motivations for attending the parties were "to listen to music and dance" and "to be with friends." CONCLUSIONS: Intensive, targeted health promotion efforts are needed for gay and bisexual men who attend circuit parties. PMID- 11392941 TI - Gonorrhea in the HIV era: a reversal in trends among men who have sex with men. AB - OBJECTIVES: Gonorrhea cases among men who have sex with men (MSM) declined in the early years of the HIV epidemic. We evaluated more recent trends in gonorrhea among MSM through the Centers for Disease Control and Prevention's Gonococcal Isolate Surveillance Project. METHODS: Isolates and case information were collected from 29 US sexually transmitted disease (STD) clinics. Gonococcal urethritis cases among MSM were compared with those among heterosexual men, and cases among MSM in 1995 to 1999 were compared with earlier MSM cases. RESULTS: Of 34,942 cases, the proportion represented by MSM increased from 4.5% in 1992 to 13.2% in 1999 (P < .001). Compared with heterosexuals, MSM were older, more often White, and more often had had gonorrhea previously, although fewer had had gonorrhea in the past year. MSM with gonorrhea in 1995 to 1999 were slightly older than those with gonorrhea in 1992 to 1994, and a higher proportion had had gonorrhea in the past year. CONCLUSIONS: MSM account for an increasing proportion of gonococcal urethritis cases in STD clinics. Given recent evidence that gonorrhea may facilitate HIV transmission, these trends demand increased attention to safe sexual behaviors and reducing STDs among MSM. PMID- 11392942 TI - Two decades after vaccine license: hepatitis B immunization and infection among young men who have sex with men. AB - OBJECTIVES: This study investigated hepatitis B immunization coverage and the extent of hepatitis B virus (HBV) infection among young men who have sex with men (MSM), a group for whom hepatitis B vaccine has been recommended since 1982. METHODS: We analyzed data from 3432 MSM, aged 15 to 22 years, randomly sampled at 194 gay-identified venues in 7 US metropolitan areas from 1994 through 1998. Participants were interviewed, counseled, and tested for serologic markers of HBV infection. RESULTS: Immunization coverage was 9% and the prevalence of markers of HBV infection was 11%. HBV infection ranged from 2% among 15-year-olds to 17% among 22-year-olds. Among participants susceptible to HBV infection, 96% used a regular source of health care or accessed the health care system for HIV or sexually transmitted disease testing. CONCLUSIONS: Despite the availability of an effective vaccine for nearly 2 decades, our findings suggest that few adolescent and young adult MSM in the United States are vaccinated against hepatitis B. Health care providers should intensify their efforts to identify and vaccinate young MSM who are susceptible to HBV. PMID- 11392943 TI - Behavioral risk factors for disease and preventive health practices among lesbians. AB - OBJECTIVES: This study compared the prevalence of health behaviors among lesbians and in the general population of women. METHODS: We used a cross-sectional community-based survey of 1010 self-identified lesbians 18 years or older. RESULTS: Compared with the general population of women, lesbians were more likely to report cigarette use, alcohol use, and heavy alcohol use. A higher percentage of lesbians were categorized as overweight, and lesbians were more likely to participate in vigorous physical activity. They were less likely to report having had a Papanicolaou test within the past 2 years but more likely to report ever having had a mammogram. CONCLUSIONS: While there may be differences in health behaviors between lesbians and the general population of women, how these differences influence the risk of subsequent disease is unknown. PMID- 11392944 TI - Patterns of cigarette smoking and alcohol use among lesbians and bisexual women enrolled in a large health maintenance organization. AB - OBJECTIVES: This study compared the prevalence of cigarette smoking and alcohol use among lesbians and bisexual women with that among heterosexual women. METHODS: Logistic regression models were created with data from an extensive member health survey at a large health maintenance organization. Sexual orientation was the primary predictor, and alcohol consumption and cigarette smoking were outcomes. RESULTS: Lesbians and bisexual women younger than 50 years were more likely than heterosexual women to smoke cigarettes and drink heavily. Lesbians and bisexual women aged 20 to 34 reported higher weekly alcohol consumption and less abstinence compared with heterosexual women and older lesbians and bisexual women. CONCLUSIONS: Lesbians and bisexual women aged 20 to 34 years are at risk for alcohol use and cigarette smoking. PMID- 11392945 TI - Health-related characteristics of men who have sex with men: a comparison of those living in "gay ghettos" with those living elsewhere. AB - OBJECTIVES: This study investigated the limitations of probability samples of men who have sex with men (MSM), limited to single cities and to the areas of highest concentrations of MSM ("gay ghettos"). METHODS: A probability sample of 2881 MSM in 4 American cities completed interviews by telephone. RESULTS: MSM who resided in ghettos differed from other MSM, although in different ways in each city. Non ghetto-dwelling MSM were less involved in the gay and lesbian community. They were also less likely to have only male sexual partners, to identify as gay, and to have been tested for HIV. CONCLUSIONS: These differences between MSM who live in gay ghettos and those who live elsewhere have clear implications for HIV prevention efforts and health care planning. PMID- 11392947 TI - HIV and sexually transmitted infection risk behaviors among men seeking sex with men on-line. PMID- 11392946 TI - Predictors of accidental fatal drug overdose among a cohort of injection drug users. AB - OBJECTIVES: This study evaluated factors associated with accidental fatal drug overdose among a cohort of injection drug users (IDUs). METHODS: In a prospective cohort study of 2849 IDUs in King County, Washington, deaths were identified by electronically merging subject identifiers with death certificate records. Univariate and multivariate Cox regression analyses were performed to identify predictors of overdose mortality. RESULTS: Thirty-two overdoses were observed. Independent predictors of overdose mortality were bisexual sexual orientation (relative risk [RR] = 4.86; 95% confidence interval [CI] = 2.30, 13.2), homelessness (RR = 2.30; 95% CI = 1.06, 5.01), infrequent injection of speedballs (RR = 5.36; 95% CI = 1.58, 18.1), daily use of powdered cocaine (RR = 4.84; 95% CI = 1.13, 20.8), and daily use of poppers (RR = 22.0; 95% CI = 1.74, 278). CONCLUSIONS: Sexual orientation, homelessness, and drug use identify IDUs who may benefit from targeted interventions. PMID- 11392948 TI - Removing the barriers: improving practitioners' skills in providing health care to lesbians and women who partner with women. PMID- 11392950 TI - Evolving medicine. PMID- 11392949 TI - Satisfaction with mental health services among sexual minorities with major mental illness. PMID- 11392951 TI - Rhesus isoimmunization. AB - Rh isoimmunization is a potentially preventable condition that occasionally is associated with significant perinatal morbidity or mortality. Disease severity may be assessed using the modalities described above and frequently, invasive techniques are required to determine the risk of severe disease. Doppler flow studies appear to offer accurate, noninvasive means of evaluating fetal risk, which may allow for a decrease in invasive diagnostic procedures. The Rh isoimmunized patient, managed by an experienced team, can anticipate a favorable pregnancy outcome. PMID- 11392952 TI - Alpha-thalassemia major: antenatal diagnosis and management. PMID- 11392953 TI - Fetal obstructive uropathy: diagnosis and management. PMID- 11392954 TI - The fetus with an abdominal wall defect. PMID- 11392955 TI - Fetal cardiac arrhythmias: diagnosis and management. AB - The diagnosis and management of fetal cardiac arrhythmias requires complex skills and knowledge, and has had a great impact on the care of infants with congenital heart disease and their families. Optimal benefits will be derived from a thoughtful team approach, with skillful internal communication, and especially when parental involvement is encouraged in the decision making process. PMID- 11392956 TI - Contemporary evaluation and management of twin-twin transfusion syndrome. PMID- 11392957 TI - Fetal surgery. PMID- 11392958 TI - Images in Medicine. Subclavian steal syndrome diagnosed by MR angiography. PMID- 11392959 TI - Prevention of pneumococcal disease among adults. PMID- 11392960 TI - Trends in inpatient cholecystectomies, 1991-2000. PMID- 11392961 TI - Identifying bacterial agents of bioterrorism: the pivotal role of the laboratory response network .2. PMID- 11392962 TI - Perspectives. Will HCFA remain a four-letter word? PMID- 11392963 TI - [Interaction of tick-borne encephalitis virus RNA with proteins]. AB - Binding of tick-borne encephalitis (TBE) virus RNA to proteins was studied by 3 methods: gel retardation, RNA-protein cross-linking under UV light, and RNA protein blotting. Two cellular proteins with molecular weights about 30 and 22 kDa were detected in the nuclear fraction of two cell strains (PS and RH) and in blood mononuclear cells of patients with TBE. Weak interaction between the studied RNA and viral proteins can result from trace amounts of TBE virus proteins in infected cells and by lack of additional binding factors. Bacterial super-products and affinity chromatography for isolation of native glycoproteins were used to increase the amount of viral proteins. Purified E and NS1 proteins did not react with viral RNA, while incubation with recombinant nonstructural TBE virus NS3 protein led to a shift in the mobility of TBE virus RNA in gel in the absence of cross-linking under conditions of UV exposure after addition of heparin. Poor binding of nonstructural TBE virus NS3 protein is based on electrostatic binding. Oligoribonucleotide 14 nucleotide residues long corresponding to 5'-terminal of TBE virus genome did not react with NS3 protein, probably because of its small size or absence of certain sequences. PMID- 11392964 TI - [Persistence of type 1 human immunodeficiency virus (HIV-1) in MT-4 cells in presence of alpha-interferon]. AB - The development of HIV infection during serial passages of HIV-1 strains with different replication capacity was studied in the presence of natural human alpha interferon (IFN-alpha). The virus activity was evaluated by accumulation of virus specific proteins in cells and culture fluid, determined by Western blot method and analysis of viral DNA in the cells by polymerase chain reaction. IFN-alpha suppressed replication of HIV-1/IIIB and 1974. The replication of strain 1974 (with a 10-fold lower replicative activity than strain IIIB) was inhibited during the first passage and of strain IIIb during the second passage. After the fourth passage IFN-2 alpha completely suppressed the replication of strain 1974, and the virus activity did not manifest after 4 consecutive co-culturings of cells from fourth viral passage with intact cells without IFN-alpha. A population of viral particles whose replicative activity was virtually undetected in the presence of IFN-alpha seemed to persist during serial passages of strain IIIB in MT-4 cells. On the other hand, the virus replication was restored during the first co culturing of cells from the virus subculture with intact cells in the absence of IFN-alpha. Presumably, an increase in the level of IFN-alpha in the blood of HIV infected patients at late stages of AIDS is not accidental and is caused by appearance of more virulent variants of the virus. PMID- 11392965 TI - [Detection of antibodies to the nucleocapsid protein of Hepatitis C virus by immunoenzyme analysis using synthetic peptides of nucleocapsid N-terminal part]. AB - Solid-phase synthesis of 7 linear peptides overlapping the immunodominant N terminal region of hepatitis C virus (HCV) core protein (aa 7-75) was carried out. Their antigen reactivity was studied by non-competitive ELISA with sera from patients with chronic HCV infection. Three B-epitopes located within aa 7-19, 20 34, and 39-75 appeared to be the most immunoreactive. Use of the set of synthetic peptides in ELISA detected anti-HCV core antibodies with 93% efficiency in comparison with ELISA and commercial anti-HCV Recomblot based on the use of commercial recombinant HCV core protein. PMID- 11392966 TI - [Dynamic of immune response in primary cytomegalovirus infection and after reactivation of cytomegalovirus in patients with organ allotransplantation]. AB - A total of 44 serum specimens from 7 patients with kidneys or liver transplanted from donors who had antibodies (Ab) to human cytomegalovirus (CMV) were studied. In 4 recipients anti-CMV Ab were found before transplantation and in 3 others they were not detected. It was shown by EIA that IgM and IgG anti-CMV appeared in the sera of primarily infected patients after 1-2 weeks and their titers were 5 10 times lower than in patients with reactivated CMV infection. Immunoblotting of Ab to individual CMV proteins showed a narrower spectrum of Ab during the initial period of primary CMV infection in comparison with the same period of reactivation and delayed production of AB to conformation-dependent determinants. Hence, analysis of anti-CMV Ab during the first 4-6 weeks after organ transplantation by EIA and immunoblotting differentiates primary CMV infection from its reactivation by Ab titers and spectrum. These parameters vary in some patients. PMID- 11392967 TI - [Dynamic of DNAse activity induction in serum of normal animals after heterogeneous DNA injection and in hereditary pathology]. AB - Studies of reactions in animals with hereditary diseases (Sapphire minks highly sensitive to Aleutian disease virus, ADV; CBA mice with 60-70% incidence of tumors; AKR mice with 90% incidence of leukemia) showed that serum DNAse activity in these animals dropped after injection of a foreign heterogeneous DNA and remained decreased during 72 h. By contrast, serum DNAse activity considerably and persistently increased after injection of DNA in Standard minks resistant to ADV, C57BI/6J mice with 1% tumor incidence, and random-bred albino mice. Presumably the capacity of standard minks to react to a foreign heterogeneous DNA by increase of DNAse activity ensures their resistance to DNA-containing ADV, while incapacity of Sapphire minks to respond to DNA by DNAse activity induction makes them sensitive to ADV. A similar relationship between the capacity to react to DNA by changes in serum DNAse activity and capacity to inherit a disease was detected in mouse strains. PMID- 11392968 TI - [Anti-Coxsakie B viruses humoral immunity status in patients with pancreatic diseases]. AB - Similar titers and incidence of antibodies to group-specific Coxsackie B virus antigen were detected in 137 patients with chronic pancreatitis and 127 patients with insulin-dependent diabetes mellitus (IDDM). Characteristic clinical features of chronic pancreatitis associated with the presence of antibodies to Coxsackie B virus in high titers were detected. HLA Cw2 antigen is significantly more incident in the phenotype of patients with chronic pancreatitis and IDDM and high titers of antibodies to Coxsackie B virus. These data confirm the possible contribution of Coxsackie B virus infection to the development of IDDM and suggest its contribution to development of chronic pancreatitis. PMID- 11392969 TI - [Status of interferon in genital infections]. AB - The interferon status test characterizes the interferon (IFN) system function and the functional activity of IFN-producing cytokine cells. In contrast to the routine method, we used fetal calf (FCS) and autologous sera for evaluating the patients' leukocyte capacity to produce IFN-alpha and IFN-gamma. Blood samples from 30 women with genital infections caused by herpes simplex virus, cytomegalovirus, Chlamydia, and Ureaplasma were tested. Autologous sera of 40% patients inhibited and of 6.6% patients stimulated the production of IFN-gamma in comparison with FCS. All the data are statistically reliable. 20.3% autologous sera contained acid-labile IFN-alpha. These results can be used for more effective immunocorrective therapy. PMID- 11392970 TI - [Phylogenetic analysis of swine fever virus strains]. AB - Amplification of H-gene fragment in combination with cDNA nucleotide sequencing can be used for indication and strain differentiation of classical swine fever virus. PMID- 11392971 TI - [Use of liposomes for vaccines design]. AB - Reviews reports on the creation of liposomal vaccines published during recent 25 years. Describes methods of antigen and immunomodulator incorporation in liposomes and their immunogenic properties. Presents data on development of influenza vaccines and prospects for their utilization as universal carriers of viral and bacterial antigens. PMID- 11392973 TI - [Genetic analysis of Japanese encephalitis virus strains and variants from Russian collection ]. AB - Genetic analysis of strains and variants of Japanese encephalitis virus from the collection of D. I. Ivanovsky Institute of Virology revealed marked differences in the complex of genetic markers, which makes possible more precise selection of the most interesting clones. PMID- 11392972 TI - [Immunological and functional characteristics of epitopes and regions of gB glycoprotein of Aujeszky's disease virus]. AB - The role of different gB epitopes and regions at some stages of virus replication in cell cultures and in the formation of immunity to Aujeszky's disease virus (ADV) was studied using a panel of 13 monoclonal antibodies (MAB) that recognize glycoprotein gB (gB) of ADV and antisera against fusion recombinant proteins expressing gB fragments. Productive infection following virion attachment was prevented by antibodies to the N-terminal domain of gB. Three MABs against the N terminal domain of gB and 5 MABs directed against the immunodominant region located in the gBc-subunit of gB inhibited the cell-to-cell spread of viral infection. After immunization with recombinant proteins expressing the N-terminal fragments of gB 80% mice were protected from lethal ADV challenge. After passive immunization the majority of MABs protected 20-80% mice from lethal ADV challenge. Hence, the N-terminal domain of ADV gB is associated with the virus penetration into the cell and is important for anti-ADV immunity. PMID- 11392974 TI - [Rabies and its prevalence in the world]. AB - Analyzes the latest reports on epidemiological situation with regard to rabies in the world and the probable causes of increase in rabies epizooties in the populations of wild and domestic animals during recent years. PMID- 11392975 TI - Cholesterol check. PMID- 11392976 TI - Personal fortune. Spa wars. PMID- 11392977 TI - They call it La-Z-Boy for a reason. PMID- 11392978 TI - U.S. faces protein deficiency. PMID- 11392979 TI - Implicit association measurement with the IAT: evidence for effects of executive control processes. AB - It is argued that a model of goal-independent spreading activation in a social or semantic knowledge structure is insufficient to explain implicit association effects in the IAT (Greenwald, McGhee, & Schwartz, 1998). An alternative account is proposed, which attributes IAT effects to differential costs for switching between task sets. Two experiments were conduced to test this account. In Experiment 1, specific task-set switching cost was a function of IAT condition: switching between tasks was associated with significantly more cost in the incompatible IAT phase. In a second experiment the magnitude of the IAT effect was reduced when task-set reconfiguration was possible in advance of or simultaneously with the upcoming stimulus. The results are discussed with respect to recently suggested accounts of the effect. PMID- 11392980 TI - Items' cross-category associations as a confounding factor in the Implicit Association Test. AB - The introduction of the Implicit Association Test (IAT; Greenwald, McGhee, & Schwartz, 1998) has stimulated numerous research activities. The IAT is supposed to measure the degree of association between concepts. Instances have to be assigned to these concepts by pressing appropriate keys as quickly as possible. The reaction time difference between certain conditions, termed the IAT effect, is used as an indicator of the degree of the concepts' association. We tested the hypothesis that the degree of association between one concept (or category) and the instances of the other presented concept also influences reaction times. In our experiment, the instances in the target categories, male and female names, were kept constant. The adjectives in the evaluative categories were manipulated: Either the pleasant adjectives were female-associated and the unpleasant adjectives were male-associated, or vice versa. These stereotypic associations were indeed found to exert a substantial influence on the size of the IAT effect. This finding casts doubt on the assumption that the IAT effect may be interpreted as a pure measure of the degree of association between concepts. PMID- 11392982 TI - Implicit attitudes towards homosexuality: reliability, validity, and controllability of the IAT. AB - Two experiments were conducted to investigate the psychometric properties of an Implicit Association Test (IAT; Greenwald, McGhee, & Schwartz, 1998) that was adapted to measure implicit attitudes towards homosexuality. In a first experiment, the validity of the Homosexuality-IAT was tested using a known group approach. Implicit and explicit attitudes were assessed in heterosexual and homosexual men and women (N = 101). The results provided compelling evidence for the convergent and discriminant validity of the Homosexuality-IAT as a measure of implicit attitudes. No evidence was found for two alternative explanations of IAT effects (familiarity with stimulus material and stereotype knowledge). The internal consistency of IAT scores was satisfactory (alpha s > .80), but retest correlations were lower. In a second experiment (N = 79) it was shown that uninformed participants were able to fake positive explicit but not implicit attitudes. Discrepancies between implicit and explicit attitudes towards homosexuality could be partially accounted for by individual differences in the motivation to control prejudiced behavior, thus providing independent evidence for the validity of the implicit attitude measure. Neither explicit nor implicit attitudes could be changed by persuasive messages. The results of both experiments are interpreted as evidence for a single construct account of implicit and explicit attitudes towards homosexuality. PMID- 11392981 TI - How robust is the IAT? Measuring and manipulating implicit attitudes of East- and West-Germans. AB - We investigated consequences of priming East-West-German related self-knowledge for the strength of implicit, ingroup-directed positive evaluations among East- and West-Germans. Based on previous studies we predicted opposite effects of self knowledge priming for East- and West-Germans. Since in general the East-German stereotype is regarded as more negative than the West-German one, bringing to mind East-West-related self-knowledge (relative to neutral priming) was expected to attenuate ingroup favoritism for East-Germans, but to increase it for West Germans. After having fulfilled the priming tasks, participants worked on an IAT version in which the to be classified stimuli were East- or West-German city names (dimension 1) and positive or negative adjectives (dimension 2). Results of Experiment 1 showed (a) that East- and West-German students implicitly evaluated their ingroups as more positive than the outgroups and (b) confirmed the predictions of the priming influence. Experiment 2 replicated these findings with more representative samples from East- and West-Germany. The results are discussed with regard to underlying processes of implicit attitudes in intergroup contexts. PMID- 11392983 TI - Heart versus reason in condom use: implicit versus explicit attitudinal predictors of sexual behavior. AB - We test the hypothesis that explicit and implicit measures of attitudes would differentially predict deliberate versus spontaneous behavior in the domain of condom use. Students completed explicit attitudinal and thought-listing measures about using condoms and implicit measures using attitude priming and Implicit Association Test (IAT) procedures. An attitude IAT measured the association between condom images and affective images; a self-identity IAT measured association of condoms with the self. We predicted and found that condom use with main partners was predicted by explicit measures but not implicit measures; the opposite was true for condom use with casual partners. Although the attitude priming measure was not positively correlated with casual condom use, the IATs were. The patterns of relations, however, were unexpectedly complex, due to a strong decrease in IAT effects over time, and different IATs assessing unique attitudinal dimensions. PMID- 11392984 TI - Attitude measurement using the Implicit Association Test (IAT). AB - Three years ago, Greenwald, McGhee, and Schwartz (1998) presented a new method to measure differential evaluative association of two target concepts: the Implicit Association Test (IAT). It has been asserted that the IAT allows for the assessment of implicit attitudes by comparing response times in two combined discrimination tasks. Although the distribution and employment of this method has been quite successful, the processes underlying IAT-effects, as well as the psychometric properties of specific IAT-variants, have received relatively little attention up to now. The articles included in the present special issue especially focus on these aspects. PMID- 11392985 TI - Health of the Implicit Association Test at age 3. AB - Since its first publication in 1998, the Implicit Association Test (IAT) has been used repeatedly to measure implicit attitudes and other automatic associations. Although there have also been a few studies critical of the IAT, there now exists substantial evidence for the IAT's convergent and discriminant validity, including new evidence reported in several of the articles in this special issue. IAT attitude measures have often correlated only weakly with explicit (self report) measures of the same associations. It therefore seems appropriate to conclude that the IAT assesses constructs that are often (but not always) distinct from the corresponding constructs measured by self-report. PMID- 11392986 TI - Figure-ground asymmetries in the Implicit Association Test (IAT). AB - Based on the assumption that binary classification tasks are often processed asymmetrically (figure-ground asymmetries), two experiments showed that association alone cannot account for effects observed in the Implicit Association Test (IAT). Experiment 1 (N = 16) replicated a standard version of the IAT effect using old vs. young names as target categories and good and bad words as attribute categories. However, reliable compatibility effects were also found for a modified version of the task in which neutral words vs. nonwords instead of good vs. bad words were used as attribute categories. In Experiment 2 (N = 8), a reversed IAT effect was observed after the figure-ground asymmetry in the target dimension had been inverted by a previous go/nogo detection task in which participants searched for exemplars of the category "young." The experiments support the hypothesis that figure-ground asymmetries produce compatibility effects in the IAT and suggest that IAT effects do not rely exclusively on evaluative associations between the target and attribute categories. PMID- 11392987 TI - [Integrated disease control in dairy herds. A case study from the veterinarians' viewpoint]. AB - Integrated control of bovine virus diarrhoea virus, bovine herpesvirus-1, Leptospira interrogans serovar hardjo subtype hardjobovis, Mycobacterium avium subsp. paratuberculosis, and Salmonella dublin in dairy herds may provide economic benefits superior to those obtained by sequential disease control, because, among other things, it allows optimization of voluntary culling. However, in practice there are no adequate instruments to establish priorities in voluntary culling. Therefore, in this study the priorities in decision-making for voluntary culling of infected cattle, as indicated by more than 300 cattle veterinarians, were analysed. Based on our results and supplementary considerations, the priorities for voluntary culling in the Netherlands can be ranked as: 1st. cull S. dublin carriers, 2nd. cull persistently infected BVDV carriers, 3rd. cull paratuberculosis faecal culture positive cattle and their last offspring, 4th. cull, in paratuberculosis infected herds, paratuberculosis ELISA positive cattle and their last offspring and cull, in low prevalence herds, BHV1 gE-positive cattle, and 5th. cull leptospirosis seropositive cattle. Since this ranking was based on one case study only, other priorities may prevail in other herds. PMID- 11392988 TI - [Equine protozoal myeloencephalitis in the Netherlands? An overview]. AB - Equine protozoal myeloencephalitis (EPM) was diagnosed in a Dutch Warmblood gelding a few months after its export to the United States. The horse came back and was treated here. Additionally, an overview of the disease complex 'EPM' is given. Mode of infection, diagnosis of disease and its differential diagnoses, and general therapeutic options are presented. Although EPM due to infection with Sarcocystis neurona in Europe seems restricted to those horses that return or are imported from North America, the possibility of future cases of EPM caused by an infection with Neospora spp. is briefly discussed. PMID- 11392989 TI - [Who is the bitten dog? Legal liability for damage done by animals. Part 3]. PMID- 11392990 TI - [Delayed issue of 'Highlights 2000' more newsworthy than ever. Dr. H.S. Frenkel, a pioneer from the early days of foot-and-mouth disease control]. PMID- 11392991 TI - [Petition from the Royal Dutch Society for Veterinary Medicine. Presented to the Minister of Agriculture, Mr. L.J. Brinkhorst, on April 17, 2001]. PMID- 11392992 TI - [Also for companion animal veterinarians: foot and mouth disease]. PMID- 11392993 TI - [A veterinarian stays a veterinarian]. PMID- 11392994 TI - [Modern communication, what use is it to veterinarians?]. PMID- 11392995 TI - [Social skills of the veterinarian in the foot and mouth disease crisis]. PMID- 11392996 TI - [Vaccine policy requirements in the Netherlands]. PMID- 11392997 TI - [Scientifically and/or economically justified?]. PMID- 11392998 TI - [Lack of civilization concerning etiology of foot and mouth disease]. PMID- 11392999 TI - [Foot and mouth disease outbreak compels one to reconsider]. PMID- 11393000 TI - [Television news]. PMID- 11393001 TI - [Quantitative magnetic resonance tomography in diagnosis of Alzheimer dementia]. AB - Alzheimer's disease (AD) is a degenerative dementing disorder which is characterised by a progressive atrophy of several brain regions. This process may be visualised in vivo by the use of magnetic resonance imaging (MRI) in combination with appropriate volumetric post-processing techniques. Recent volumetric MRI studies in AD consistently found an extensive volume loss of the medial temporal lobe structures including amygdala and hippocampus which appeared already in the early clinical stages of the disorder. This finding is progressive during the clinical course of AD and is associated with other biological markers of the disease such as cerebrospinal fluid beta A 4 levels and apolipoprotein E genotype. With respect to the extent and the distribution of the structural changes, AD may be differentiated from other neuropsychiatric disorders which could facilitate the differential diagnosis in vivo. PMID- 11393002 TI - [Dementia in advanced age: estimating incidence and health care costs]. AB - Based on results from large-scale epidemiological field studies in the western industrial countries, 930,000 elderly people in Germany were estimated to suffer from a dementing disorder at the end of 1996. Following the most recent population projection, a population increase of the number of elderly people (65 yrs. and above) from 12.9 million (mio.) in 1996 to more than 20 mio. in 2030 is anticipated. Based on the assumption that age-specific prevalence rates of dementia will remain stable, a steep rise in patient numbers by an average of 20,000 per year can thus be expected, reaching 1.56 mio. in 2030 and more than 2 mio. in 2050. Studies on the cost of illness point to an enormous economic burden caused by dementia. The unpaid informal care provided by relatives and the high expenses for long-term institutional care can be considered as the most significant components of total costs. Currently, the medical costs associated with diagnosis and treatment, however, appear as an almost negligible fraction of the total costs. PMID- 11393003 TI - [Non-medicamentous therapy and prevention of Alzheimer disease]. AB - An overview of prevalence data and empirically documented risk factors of a dementia is followed by a short description of intervention strategies. After this, prevalence data from the SIMA Study are presented and results of the multivariate analyses of early indicators and risk factors are discussed. The last section is dedicated to the risk decreasing effects of a combined memory and psychomotor training program which was developed within the study's scope. PMID- 11393004 TI - [Prolonging life at any cost? Ethical attempts at new personal freedom regarding and and the end of life]. AB - The prolongation of life to its extremes "at any cost" has been criticized for decades. The courts have endorsed patient autonomy and patient rights. The number of advance directives has increased considerably. The medical community did after all react appropriately, e.g., by codes of ethics or guidelines. The situation is now in danger of overreaction. The problems of lopsided self-determination are at stake. A new culture of negotiated medical constraint needs both doctor and patient as partners ready to learn how to walk the narrow ridge between overtreatment and just individual choice. PMID- 11393005 TI - [Gerontology and nursing care]. AB - This paper focuses on questions of the philosophy of science and the scientific development of gerontology and nursing science. First, some aspects of the scientific development in gerontology and nursing science (autonomy, inter- and multidisciplinarity as well as the theory debates) are summarized. In gerontology, problems of the philosophy of science are often neglected. The main focus is on empirical research and the establishment of an scientific infrastructure. In nursing science are questions of the philosophy of science, especially debates about the discipline, are significant. In Germany nursing research and the establishment of a scientific infrastructure are still in the initial stages. Second, on the basis of a content analysis the relevance and status of "nursing/need of care/frailty" in two leading journals ("Zeitschrift fur Gerontologie und Geriatrie" and the "Pflege") is shown. Main result is that in nursing science publications regarding the status of the discipline, studies of the motivation, attitudes and behavior of nurses as well as investigations of the professionalization are dominant. In gerontology, the main emphasis is on studies of the changing health service structure, geriatric assessment and qualification. Finally chances of an interdisciplinary exchange between gerontology and nursing science are discussed. PMID- 11393006 TI - [Consumer perspective and quality development in ambulatory nursing care]. AB - In the 1990s, the debate surrounding the issue of quality in outpatient nursing care was given a remarkable boost. Also the conviction steadily grew that quality assurance should be more adjusted to values and expectations of people in need of nursing care, the addressees of such services. But a critical analysis is required to determine to what extent the role of "customer" or "consumer", a role often ascribed to them in this context, can actually be reconciled with their possibilities, expectations and interests. Based on the results of empirical studies, the following article shows that the patient's view, his expectations and evaluations largely depend on some specific consequences of being in need of care and using outpatient nursing care. The article goes into the divergent perspectives that, on the one hand, are characteristic of patients and, on the other, of the nursing staff and other bodies concerned with quality of care. PMID- 11393007 TI - [Classical cardiovascular risk factors: predictive value and treatment of the elderly. The rocky road to evidence-based medicine]. AB - Coronary heart disease and cerebrovascular disease are still the most common causes of death in Western countries. A number of risk factors have been identified in young and middle-aged adults, such as dyslipidemia, hypertension and diabetes. Their prevalence and importance, however, are less clear in the elderly. In terms of dyslipedemia it is questionable whether hypercholesterolemia is a definite risk factor. On the other hand, mortality can be reduced by lowering LDL cholesterol, but the benefit in the oldest old is not yet known. Systolic blood pressure rises with age and is discussed controversely as a potential risk factor in the elderly. Some large trials could show a clear relationship between high blood pressure while others did not see any association. Similar to the treatment of hypercholesterolemia, antihypertensive drugs showed beneficial effects in elderly people until the age of 80. But the treatment of the oldest old cannot be recommended in general. Diabetes and impaired glucose tolerance are some of the most common diseases in elderly people. They are considered to be an important risk factor until the age of 75. Their role in the oldest old is still under debate. Until now, we do not know anything about possible treatment effects because of the lack of controlled trials. Elderly people seem to have a risk profile different from younger people; especially in extreme ages the predictive role of classical risk factors is unclear. On the other hand, drug treatment could reduce mortality and morbidity in patients with hypercholesterolemia or hypertension. There are no studies which investigated the effects of blood glucose control in the elderly. The collection of sufficient data is a geriatric challange in order to decide whether treatment is useful or not. PMID- 11393008 TI - Management of urinary tract infections in the elderly. AB - Urinary tract infection (UTI) is the most common infection and the first cause of bacteremia in the elderly. With increasing age the female to male ratio decreases and UTI becomes almost half as frequent in men compared to women. Significant bacteriuria exists in about 40% of institutionalized women. But asymptomatic bacteriuria is neither the cause of morbidity nor associated with a higher mortality rate and thus should not be treated. Symptomatic infection in women without complicating factors is most often caused by E. coli and may be treated with 3 or 7 day regimens of trimethoprim-sulfamethoxazole or fluoroquinolones (FQ). In the presence of symptoms of upper tract infection or complicating factors, urine culture is mandatory and will detect multiple and/or resistant microorganisms in most cases. Empirical treatment has to be adapted according to the sensitivity once established and should be administered for at least 10 days. Most of the patients above 65 and virtually all patients above 80 present either with general debility or diabetes or other factors such as bladder outflow obstruction or abnormal bladder function and have to be considered as presenting with complicated UTI. Indwelling catheters should be removed if possible, otherwise be changed. PMID- 11393009 TI - [5th Congress of the German Society of Gerontology and Geriatrics]. PMID- 11393010 TI - [Socio- and psychotherapy in patients with Alzheimer disease]. AB - Symptoms presented by patients with Alzheimer-type dementia do not only reflect organic disturbances only but require a holistic and person-oriented view. Affective and behavioral disturbances are not necessarily secondary to cognitive impairment. Guidelines are presented for a multidimensional treatment involving the significant other. Socio- and psychotherapy are essential for this treatment. Their approaches have greatly increased in number and diversity in the past few years. Sociotherapy is based on milieu therapy and includes different training- and group activities. Several psychosocial treatment modalities are available, including validation, dementia care mapping, reminiscence therapy, cognitive training and psychoeducational group work. Psychotherapeutic approaches include relaxation techniques, and psychodynamic oriented- and behavioral modalities. The indication for a specific modality is based on an assessment of the disturbances present and available resources. Of special importance are also services to family carers, including counseling, psychotherapy, as well as support and modification of the care-setting. Even though there are only limited empirical data available on the effects of socio- and psychotherapy for patients with Alzheimer-type dementia, the available evidence is indicative of a positive influence on symptoms of this illness. Diversity of symptoms and individualized, variable course of the illness may point to the importance of psychological and social factors in this illness, by far larger than presently recognized. PMID- 11393011 TI - Automatic utilities auditing. PMID- 11393012 TI - Correct design can deliver double payoff. PMID- 11393013 TI - Evaluating the costs of pollution from construction. PMID- 11393014 TI - Service comprehensively upgraded. PMID- 11393015 TI - Waiting lists. Bone of contention. AB - The number of orthopaedic consultants has increased by 78 per cent since 1984, but average waiting time has hardly changed. Orthopaedics is one of the greatest problem areas for waiting lists although workloads have fallen by about a fifth in the past five years. An extra 85,000 orthopaedic patients will have to be admitted over the next five years if the government is to eliminate waits of more than six months. Consultants oppose the expansion of the staff grades. Specialist centres may have to be established to deal with the backlog. PMID- 11393016 TI - Helplines. Getting through. AB - A specialised cancer telephone advice line deals with 1,000 calls a week. But many callers don't get through first time. The helpline operates Monday to Friday, 9am-7pm. Peaks in demand follow television and newspaper coverage of cancer. The launch of a website has not led to a decrease in calls. Callers show a high degree of persistence. PMID- 11393017 TI - [About education of specialists in clinical laboratory diagnosis in the Voronezh State University]. PMID- 11393018 TI - [Organization of a course of clinical laboratory diagnosis (clinical biochemistry) for laboratory physicians]. PMID- 11393019 TI - [New forms of optimizing the laboratory process (cooperation of theoretical and clinical departments)]. PMID- 11393020 TI - [Continuing education of specialists in clinical laboratory diagnosis]. PMID- 11393021 TI - [Some problems in specialists training for clinico-diagnostic laboratories ]. PMID- 11393022 TI - [Quality control of continuing education of specialists in clinical laboratory diagnosis]. PMID- 11393023 TI - [The new method of continuing education of students and physicians in biochemistry, clinical biochemistry and clinical laboratory diagnosis]. PMID- 11393024 TI - [Training of instructors for departments of clinical laboratory diagnosis]. PMID- 11393025 TI - [New potentials for laboratory analytical methods: microfluid chip analyzers (lecture)]. PMID- 11393026 TI - [Training of specialists for clinical laboratories: traditions and trends (comments of the problem coordinator in the Research Committee of the Congress)]. PMID- 11393027 TI - [Immunological aspects of laboratory diagnosis of chronic urticaria]. AB - High prevalence of allergic diseases necessitates search for new methods of laboratory diagnosis thereof. We studied the diagnostic significance of the count of cells expressing low-affinity receptors to IgE (CD23+ cells) and compared this test with skin tests with non-infectious allergens and measurement of total serum IgE. 104 patients with various forms of chronic relapsing urticaria were examined. The count of CD23+ cells was markedly increased in atopic urticaria. The increase in the count of these cells and correlation with the results of skin test were less expressed in infectious allergic urticaria. In other forms of chronic urticaria characterized by negative results of skin tests the count of CD23+ cells was normal. The level of total serum IgE was low virtually in all patients. Hence, the count of cells carrying low-affinity receptors to IgE is highly informative, detecting IgE-mediated reactions in the patients, though this parameter does not allow identification of the allergen. This test can be used in complex with other methods of allergodiagnosis, particularly in cases when skin tests are for this or that reason impossible. PMID- 11393028 TI - [Education of specialists in clinical laboratory diagnosis in the Russian Federation]. PMID- 11393029 TI - [The effective method for detection of IgG-antibodies of AB0 system and its rapid micromodification ]. AB - Presents a method for measuring AB0 antibodies (IgG) in the serum and/or colostrum with 2-mercaptoethanol pretreatment to reduce disulfide bonds in the IgM molecule. Offers a highly sensitive, rapid, economic, and effective variant of the test in polystyrene plates for immunological tests, which can be used in screening studies. PMID- 11393030 TI - [Surface relief of peripheral blood erythrocytes in patient with schizophrenia (scanning electron microscopy data)]. AB - Surface relief and configuration of peripheral blood erythrocytes were studied under a scanning electron microscope in 42 patients with paranoid schizophrenia during the active phase and remission. Control group consisted of 30 mentally and somatically healthy donors. The count of functionally intact biconcave discocytes was appreciably (p < 0.001) decreased and the incidence of transitory, prehemolytical, and degenerative forms of erythrocytes increased in the patients. Morphological restructuring of the erythrocyte population was universal in the patients with different duration of the schizophrenic process. PMID- 11393031 TI - [A case of congenital dyserythropoietic anemia in newborn]. PMID- 11393032 TI - [Study of leukocytic component of nonspecific resistance in patients with bronchopulmonary diseases living in the Aral region]. PMID- 11393033 TI - [Free radicals lipid peroxidation and level of R-proteins in non-specific aortoarteritis and hypertension]. AB - Parameters characterizing free-radical oxidation of lipids (malonic dialdehyde, total serum antioxidative activity, ceruloplasmin, and superoxide dismutase) and the level of R proteins were studied in patients with nonspecific aortoarteritis and essential hypertension. Lipid peroxidation and the content of regulatory proteins were increased in the patients, the shifts being the most expressed in nonspecific aortoarteritis of the maximum activity. PMID- 11393034 TI - [The method for detection of urinary lysozyme]. AB - A method for lysozyme measurement in the urine is developed, based on adsorption of lysozyme molecules on the surface of chitin. The urine of healthy children, men, and women contains 1.76, 1.51, and 1.69 mg/liter lysozyme, respectively. Urinary lysozyme values in some diseases are presented. PMID- 11393035 TI - [Fibronectin content in the urine of patients with chronic glomerulonephritis as a test for the efficiency of treatment]. AB - Renal fibronectin synthesis is impaired in patients with chronic glomerulonephritis. We measured urinary fibronectin for evaluating the efficiency of various methods of treatment. Traditional therapy of patients with the nephrotic syndrome at the stage of renal failure leads to decrease of fibronectinuria, which can be indicative of the progress of nephrosclerotic process in the renal parenchyma; monotherapy with wobenzyme during the azothemic stage of disease in patients with the urinary and nephrotic syndrome does not cause statistically significant changes in the level of urinary fibronectin, which can be regarded as inhibition of nephrosclerosis process. Hence, wobenzyme is the drug of choice, decreasing the velocity of nephrosclerotic processes, when pathogenetic therapy is largely limited or precluded. Combination of wobenzyme with pathogenetic drugs in patients with the nephrotic syndrome and intact renal function suppresses fibronectinuria due to mutual potentiation of the antiinflammatory effect. Decrease of fibronectin concentration in the urine after wobenzyme monotherapy in patients with the urinary syndrome without signs of chronic renal insufficiency confirms the antiinflammatory effect of the drug. PMID- 11393036 TI - [The present and future of laboratory medicine in Samara. Training problems]. PMID- 11393037 TI - A setback for medipot. PMID- 11393038 TI - New hope for cancer. PMID- 11393039 TI - Cholesterol alert. PMID- 11393040 TI - The single life. PMID- 11393041 TI - Heartsick America. PMID- 11393042 TI - Unmarried with children. PMID- 11393043 TI - Why I think I'm still right. PMID- 11393044 TI - Is it healthy for the kids? PMID- 11393045 TI - [Incidence of non-compliance in asthma patients: current research status and methodological problems]. PMID- 11393046 TI - [Therapy participation in ambulatory asthma patients: empirical comparison of compliance rates using different operationalization methods for drug compliance]. PMID- 11393047 TI - [Ethical problems of electronic compliance measurement in asthma patients]. PMID- 11393048 TI - [A new look at lung abscess. Clinical treatment-related classification]. PMID- 11393049 TI - [Diagnosis of reading and spelling disorder]. AB - The ICD-10 calls for the use of tables that account for the correlation between intelligence and spelling or reading, respectively (regression model) in the diagnosis of dyslexia. In this paper we discuss the consequences that arise from this recommendation with respect to the interpretation of psychometric tests. In addition, a table is presented that contains the data required to make diagnostic decisions based on the regression model. Furthermore, an expected prevalence rate was calculated using randomized computer data. PMID- 11393050 TI - [Differences between adolescent patients with anorexia and bulimia nervosa with reference to psychological and psychosocial markers]. AB - OBJECTIVES: We present a retrospective study of 140 patients (110 with anorexia nervosa, 30 with bulimia nervosa), hospitalized between 1982-1992 at the Department of Child and Adolescent Psychiatry at the University of Wuerzburg, Germany. METHODS: All patients met the ICD-10 criteria for anorexia nervosa or bulimia nervosa. We collected data from basic documentation and the multiaxial classification (MAS), using a variety of standard instruments such as Anis 32, MMPI, BDI, HAWIK-R and HAWIE. RESULTS: Our findings show significant differences between the two populations. At the time of their first admission anorectic patients were somewhat younger than their bulimic counterparts (14.5 years vs. 16.5 years, respectively). With regard to the typical symptoms of either disorder, the two populations differ in their eating behavior on the factor bulimia described by Anis-32. A comparison of personality features reveals that anorectic patients scored lower than bulimic patients on the MMPI scales, especially on psychopathology, but higher on depression in the BDI. Other characteristics of anorectic patients include a higher-than-average IQ, more enmeshment and overprotectiveness in family relations, more separation anxiety and insufficient communication skills at school. By contrast, bulimic patients demonstrated poorer scholastic performance and more discipline problems at school, while communication among family members was impaired. PMID- 11393051 TI - ["Nothing could have stopped me, neither God nor the world!"--case report of a 14 year-old suicide patient]. AB - We present the case of a 14-yearold girl who imitated the railway suicide of her boyfriend as a problem-solving strategy. Theories about the etiology of suicide behavior, particularly among vulnerable adolescents, are discussed. PMID- 11393053 TI - [Developmental psychopharmacology]. PMID- 11393052 TI - [Agranulocytosis in a child with schizophrenia treated with clozapine--clinical findings and therapy, a case report]. AB - OBJECTIVES: The indication of clozapine therapy is governed by special guidelines due to a 1%-3% risk of clozapine-induced agranulocytosis. Up to now there has never been a report of such a case in a child with schizophrenia. The case report presented here is concerned with the clinical features and the treatment of clozapine-induced agranulocytosis in childhood schizophrenia. METHODS: It deals with the treatment of a 12-yearold boy with a schizophrenic psychosis. The psychotic symptoms before treatment and during inpatient treatment are described. The procedures for the diagnosis and treatment of the clozapine-induced agranulocytosis are presented. RESULTS: Clozapine medication may induce agranulocytosis in the treatment of a child with a schizophrenic psychosis. The highly specific guidelines governing its use must be followed as well in the treatment of very early onset schizophrenia. An agranulocytosis may result following 15 weeks of clozapine medication. The treatment with granulocyte colony stimulating factor seems to support normalization of the blood picture. PMID- 11393054 TI - [Can differences in stress perception and coping explain sex differences in depressive symptoms in adolescents?]. AB - A four-year longitudinal study explored the different contribution of low self esteem, different types of stressors, conflict in close relationships and avoidant coping to the explanation of depressive symptomatology in adolescents. One hundred and ninety adolescents, 101 females and 89 males, participated in four annual assessments using diverse instruments. ANOVAs repeated measurements revealed a higher stress level, more conflicts with mothers and more avoidant coping in females as compared to males at the age of 14 years. Males showed fewer depressive symptoms and higher positive self-esteem at all times. Multiple regression analysis revealed that stress and avoidant coping in early and mid adolescence explained a significant proportion of depressive symptoms among females in late adolescence. Among males, only the level of conflicts with friends in early adolescence contributed to their level of depressive outcome in late adolescence. PMID- 11393055 TI - [Victimization by peers as a developmental risk factor]. AB - The phenomenon of victimization in the peer group has received considerable attention in recent years. This review examines the current state of research in this area. About 10% of all students are affected by peer victimization and are repeatedly verbally or physically attacked by one or more of their fellow students. The causes are to be sought in the social context as well as in individual deficits in social skills, but also in the social-cognitive attributional patterns of the children. The parent-child relationship is another important risk factor. The stability of victimization is relatively high. Its short-term consequences consist of a negative mood and low self-esteem. In the long-term children feel lonely, helpless and sad. Recent research underscores a connection between victimization and the development of depressive disorders. Protective factors are composure in conflict situations, and especially the availability of at least one good friend. Therapeutic interventions are to be considered at the individual level as well as at the level of the classroom unit. Training in certain areas of social competence would be helpful for the children, but consultation for their parents and teachers is important in order to interrupt certain group dynamic processes in the classroom. Future research should address the topic of peer victimization in clinical samples. PMID- 11393056 TI - Variation in the incidence of AGUS between different patient populations. AB - OBJECTIVE: To determine the frequency of atypical glandular cells of undetermined significance (AGUS) for three consecutive calendar years from three different referral sources. STUDY DESIGN: Cervicovaginal smears with a diagnosis of AGUS were identified from January 1995 through December 1997. The smears were submitted from three different sources: two were city government hospital clinics, one with predominantly African American and Hispanic patients and the other with predominantly Asian and Hispanic patients. The third referral source was private practitioners' offices with predominantly Caucasian patients. RESULTS: A diagnosis of AGUS was made in 707 cases, accounting for 0.56% of all smears examined. This was in contrast to 6,872 smears reported as atypical squamous cells of undetermined significance (ASCUS) (5.4%) and 3,347 reported as squamous intraepithelial lesions (SIL) or above (2.7%). The incidence of AGUS ranged from 0.16% to 1.00% among different patient populations. This difference was also noted in the rate of ASCUS and SIL in the same patient population. There was a steady increase in the rate of AGUS for each referral source during the study period. The overall rate of patients who underwent histologic evaluation and the incidence of biopsy-proven preinvasive and invasive lesions were 62.4% and 23%, respectively. There was no significant difference in the rate of significant lesions after a diagnosis of AGUS during the study period or between the three referral sources. CONCLUSION: The AGUS rate in our laboratory was low and within the range (0.17-1.83%) reported in the literature. The AGUS rate varies with different patient populations, particularly with the incidence of SIL and age distribution. PMID- 11393057 TI - Significance of AGUS Pap smears in pregnant and postpartum women. AB - OBJECTIVE: To study the clinical significance of atypical glandular cells of undertermined significance (AGUS) in pregnant and postpartum women. STUDY DESIGN: We evaluated 35 women who were pregnant (30) or within three months postpartum (5) and had a cytologic diagnosis of AGUS. Twenty-seven (77%) patients had follow up: 17 (63%) patients underwent colposcopic examination and biopsy, and 10 (37%) had repeat Pap smears. Eight patients were lost to follow-up. RESULTS: Five (29.4%) patients had a squamous intraepithelial lesion (SIL), including three high grade and two low grade, on subsequent biopsy. The remaining (70.6%) patients had benign pathology, which included 5 chronic cervicitis, 4 endocervical and/or endometrial polyps, 2 Arias-Stella reaction and 1 microglandular hyperplasia. Among the patients with repeat Pap smears, two had persistent AGUS/atypical squamous cells of undetermined significance, the remaining cases were within normal limits. CONCLUSION: Pregnancy-related changes may present with glandular atypia. In addition, about one-third of pregnant and postpartum women with a diagnosis of AGUS had SIL on subsequent biopsy; that rate is similar to that in nonpregnant women. Therefore, pregnant women with a cytologic diagnosis of AGUS should be followed closely. PMID- 11393058 TI - Value of fine needle aspiration cytology in the initial diagnosis of Hodgkin's disease. Analysis of 188 cases with an emphasis on diagnostic pitfalls. AB - OBJECTIVE: To evaluate the diagnostic accuracy and pitfalls of fine needle aspiration (FNA) cytology in the initial evaluation of Hodgkin's disease (HD) and to assess the influence of the pathologist's experience by comparing the results during two periods. STUDY DESIGN: A total of 170 cytodiagnoses of HD were reviewed and compared with those on the final histopathologic report. Thirty three cases of HD with a previous, different cytologic diagnosis were also selected. In all the cases under study, FNA was performed as part of the initial diagnostic approach. From a practical perspective, diagnostic errors were divided into major or minor according to the consequences on patient management. RESULTS: Fifteen cytologic diagnoses of HD were followed by a different histologic diagnosis after lymph node biopsy. In 33 cases of HD an erroneous cytologic diagnosis was given prior to biopsy. The sensitivity of the series was 82.4% (86.1% excluding nonrepresentative cases). The positive predictive value reached 91.2%. Sensitivity varied from 79.3% in the first period (1982-1990) to 84.9% in the second (1991-1999) (83.3% and 88.2%, respectively, excluding nonrepresentative cases). Similarly, the positive predictive value increased from 89% to 92.8%. Diagnostic errors with important consequences for patient management diminished from 14 in the first period to 5 in the second. CONCLUSION: Cytology offers a rapid and accurate approach not only for the diagnosis of recurrent HD but also for its initial recognition. These results increase the capacity of FNA as a first-level diagnostic technique in the screening of lymphadenopathies. PMID- 11393059 TI - Combined cytomorphologic and immunophenotypic analysis in the diagnostic workup of lymphomatous effusions. AB - OBJECTIVE: To analyze the results of cytomorphology and immunophenotyping in 54 patients with lymphomatous effusions. STUDY DESIGN: We report the results of cytomorphology and immunophenotyping in 54 patients with lymphomatous effusions. Twenty-three of the 54 had a previous diagnosis of NHL. In the remaining 31 patients, lymphomatous involvement was clinically suspected. RESULTS: Thirty three lymphomatous effusions were positive for involvement by NHL. Twenty-one of these 33 patients (64%) had a previous diagnosis of NHL. Of the remaining 12 patients with newly diagnosed NHL, 11 had high grade lymphoma, and one had follicular center lymphoma. Twenty effusions were considered to be reactive; only two of these patients had NHL. One effusion revealed involvement by a previously unknown carcinoma. We observed seven false negative results if only one of both methods was considered. A high grade NHL was not diagnosed by immunophenotyping in one case, and six cases of low grade NHL could not be detected by cytomorphology. The combined strategy of cytomorphology and immunophenotyping had a sensitivity of 100% and specificity of 100% in our study, confirmed by follow up studies. CONCLUSION: Both methods have shown difficulties in the examination of lymphomatous effusions. Cytomorphology has problems distinguishing reactive effusions from low grade NHL. The detection of high grade NHL by immunophenotyping is difficult. However, both methods together offer the advantage of dual staining ability and are most helpful in distinguishing clonal lymphomatous from reactive effusions. PMID- 11393060 TI - Opportunistic testing of medically underserved women for cervical cancer in South Africa. AB - OBJECTIVE: To determine the yield of opportunistic Pap smears taken in an unscreened and medically underserved population in the Transkei Region of South Africa. STUDY DESIGN: Cross-sectional study of 22,160 cervical cytology specimens from an unscreened population attending gynecologic outpatient clinics between January 1990 and December 1996. RESULTS: The overall prevalence of atypical squamous cells of uncertain significance (ASCUS), low grade squamous intraepithelial lesions (LSIL) and high grade squamous intraepithelial lesions (HSIL) was, respectively, 34.7%, 8.3% and 2.4%. The ASCUS: SIL ratio was 3:1. The prevalence of invasive squamous cell carcinoma was 1.6%. The yield of opportunistic Pap smears was 10.7% including only LSIL and HSIL. CONCLUSION: The pathologic process of precursor lesions of cervical cancer appears to start at an early age since > 20% of cases are diagnosed before the age of 30 years. In the absence of a national screening program, opportunistic testing of medically underserved women needs to be maintained and encouraged. PMID- 11393061 TI - Bronchoalveolar lavage fluid cytology in patients with Pneumocystis carinii pneumonia. AB - OBJECTIVE: To evaluate bronchoalveolar lavage (BAL) cytology and organism burden in patients with Pneumocystis carinii pneumonia (PCP) who were infected with the human immunodeficiency virus (HIV) and in those with other immunodeficiencies. STUDY DESIGN: BAL fluid samples from patients with PCP were selected (HIV infected patients, n = 15; patients with other immunodeficiencies, n = 11). May Grunwald-Giemsa-stained cytocentrifuge preparations were evaluated. Foamy alveolar casts (FACs) and P carinii clusters were counted. RESULTS: The numbers of FACs and P carinii clusters in BAL fluid samples of HIV-infected patients were significantly higher as compared to those in samples from patients with other immunodeficiencies. Striking cytologic findings observed in half the samples from both patient groups included the presence of foamy alveolar macrophages, activated lymphocytes, plasma cells and reactive type II pneumocytes. Furthermore, a peculiar cell type, "nonidentified cell" (NIC), was observed almost exclusively in BAL fluid samples from HIV-infected patients. CONCLUSION: BAL fluid samples from HIV-infected patients with PCP displayed higher organism burdens as compared to those from patients with other immunodeficiencies. Moreover, cytologic findings suggestive of noninfectious lung conditions were common in BAL fluid samples obtained from patients with PCP. Further study is required to elucidate the identity of the NIC cell type. PMID- 11393062 TI - Langerhans cell histiocytosis in lymph nodes. Cytomorphologic diagnosis and pitfalls. AB - OBJECTIVE: To delineate the cytomorphology of Langerhans cell histiocytosis (LCH) in lymph nodes. STUDY DESIGN: Nine histologically documented LCH cases with a prior lymph node aspirate and five more cases in which a cytologic diagnosis of LCH was rendered in a background of corroborative clinical and radiologic findings were included in a retrospective study over a 12-year period (January 1988-January 2000). Papanicolaou- and May-Grunwald-Geimsa-stained smears were reviewed by two independent observers. Staining for S-100 protein was available in four cases. RESULTS: Nine cases had multisystem involvement, while in five cases only lymph nodes were involved. The ages ranged from 5 months to 27 years, with 11 males and 3 females. An initial cytologic diagnosis of LCH had been rendered in six, suspected in four and missed in four. On review, all were reclassified as LCH except two cases, which were still thought to be reactive and necrotizing lymphadenitis. The pathognomic feature of LCH, the "LCH cell," was identified in 12 of 14 cases along with varying numbers of eosinophils, polymorphs and lymphocytes. Giant cells were seen in six cases, and plasma cells were rarely seen. CONCLUSION: Lymph node involvement by LCH can be identified by fine needle aspiration in 85% of cases. The presence of the LCH cell is a must. The differentials to be considered are dermatopathic lymphadenitis, sinus histiocytosis with massive lymph-adenopathy, Hodgkin's lymphoma and malignant histiocytosis. PMID- 11393063 TI - Polymerase chain reaction vs. conventional diagnosis in fine needle aspirates of tuberculous lymph nodes. AB - OBJECTIVE: To compare four conventional methods of diagnosing tuberculous lymphadenophathy (TL)--namely fine needle aspiration cytology (FNAC), Zeihl Neelsen staining of smears for acid-fast bacilli (AFB), culture for Mycobacterium tuberculosis (MTB) and lymph node biopsies--with the polymerase chain reaction (PCR) in order to assess the practicability and advantage of its use in routine diagnosis in a developing country. STUDY DESIGN: Fine needle aspirates from 142 consecutive patients presenting with lymphadenopathy (mainly cervical) without any known systemic involvement underwent cytomorphologic diagnosis, AFB smears, culture for MTB, confirmatory biopsy and PCR for MTB. The aspirates from cases other than TL served as controls for PCR. RESULTS: Correct diagnosis of tuberculosis could be made in 94.87% of cases by a combination of the four methods. PCR was done in 52 cases, 39 confirmed TL and 13 controls. Sensitivity, specificity, positive predictive value (PPV) and negative predictive value of PCR were 94.44%, 38.23%, 44.73% and 92.85%, respectively, when culture alone was considered the gold standard. However, specificity (38.23-92.30%) and PPV (44.73 97.36%) of PCR increased remarkably when response to treatment was taken as the final arbiter. CONCLUSION: The four conventional tests were found to be the methods of choice for the diagnosis of TL in developing countries. PCR should be reserved for problem cases. PMID- 11393064 TI - Thyroid papillary microcarcinoma. Is it really a pitfall of fine needle aspiration cytology? AB - OBJECTIVE: To assess the role of fine needle aspiration (FNA) of the thyroid in the diagnosis of papillary microcarcinoma. STUDY DESIGN: Eight cases of papillary microcarcinoma were diagnosed by fine needle aspiration. On histologic examination they were found to be adjacent to larger nodules of interest. The microcarcinomas were inadvertently sampled when sampling the larger, dominant nodules. RESULTS: None of the eight dominant nodules were papillary carcinoma; seven were benign lesions, and one was an angioinvasive Hurthle cell carcinoma. In three cases the microcarcinomas were situated within the capsule of a hyperplastic nodule. On histologic examination, five cases had multifocal microcarcinomas, with one case having multiple lymph node metastases. Based on the clinical findings and morphologic features, there were no definitive cytologic findings that could distinguish between "incidental" microcarcinoma and clinically significant papillary carcinoma. CONCLUSION: The detection of microcarcinoma by FNA should not be considered a false positive finding since the exact nature of the lesion cannot be determined until complete histologic evaluation reveals it to be truly incidental and clinically insignificant. PMID- 11393065 TI - Association of different pathologic processes of the thyroid gland in fine needle aspiration samples. AB - OBJECTIVE: To evaluate the possible significant association between different pathologic processes of the thyroid gland. STUDY DESIGN: From a series of 10,039 fine needle aspiration biopsies of the thyroid gland, a total of 1,330 cases were aspirated involving two or more palpable nodules. In 103 (1%) cases, two different pathologic processes were cytologically diagnosed. Statistical analysis was performed through four two-tailed chi 2 tests to evaluate the following events: (1) mononodularity and multinodularity vs. neoplasms, (2) colloid goiter and neoplasms, and (3) all neoplasms vs. colloid goiter and lymphocytic thyroiditis. All tests were performed using < .05 as the probability level. RESULTS: Simultaneous pathologic processes observed were: goiter and chronic lymphocytic thyroiditis (32), goiter and Hashimoto's thyroiditis (21), goiter and subacute granulomatous thyroiditis (8), goiter and follicular neoplasm (9), and goiter and papillary carcinoma (14). In 12 cases we found goiter and Hurthle cell tumor, goiter and medullary thyroid carcinoma (2), papillary thyroid carcinoma and Hashimoto's thyroiditis (2), Graves' disease and lymphocytic thyroiditis (2), and follicular neoplasm and lymphocytic thyroiditis (1). Statistical analysis showed significant association between multinodularity and neoplasms (P < .001), while the association between goiter and any type of neoplasia was not statistically significant. All the neoplasms taken together were associated with lymphocytic thyroiditis and goiter (P < .005). CONCLUSION: While there may be no statistically significant associations between the individual pathologies, it seems that having one pathology increases the risk of developing another. All the palpable nodules in the same gland should be investigated by fine needle aspiration in order to improve diagnostic sensitivity and to identify occult neoplasms. PMID- 11393066 TI - Fine needle aspiration cytology of sclerosing adenosis of the breast. AB - OBJECTIVE: To analyze the cytologic findings of sclerosing adenosis of the breast. STUDY DESIGN: We reviewed the fine needle aspiration (FNA) cytologic slides of 7 cases of sclerosing adenosis of the breast and compared the cytologic findings with those of 10 cases of fibroadenoma and 7 cases of fibrocystic change. RESULTS: The smears of sclerosing adenosis were moderately to markedly cellular, consisting of small to large groups of benign epithelial cells arranged with variable architecture. Acinar sheets, scattered individual epithelial cells and small, dense, hyalinized stroma were found in all cases of sclerosing adenosis. Epithelial cells in sclerosing adenosis appeared more frequently as acinar sheets and discohesive individual cells than did those of fibroadenoma (P < .05). However, the branching pattern of epithelial sheets, large sheets and bipolar, naked nuclei were commonly found in fibroadenoma (P < .05). Fibroadenoma had large, hypocellular, fibromyxoid stroma, whereas sclerosing adenosis had small, dense, hyalinized stroma occasionally attached to the epithelial sheets. As compared with fibrocystic change, sclerosing adenosis had similar findings but showed more abundant cellularity, acinar sheets and individual epithelial cells, and the presence of stroma (P < .05). CONCLUSION: Findings of frequent acinar sheets and small, dense, hyalinized stroma attached to epithelial sheets can aid the FNA cytologic diagnosis of sclerosing adenosis. Awareness of the presence of scattered individual epithelial cells in cytologic smears of sclerosing adenosis can help prevent a misdiagnosis of malignancy. PMID- 11393067 TI - Oral exfoliative cytology in the diagnosis of paracoccidioidomycosis. AB - OBJECTIVE: To assess the efficacy of conventional oral exfoliative cytology as a diagnostic tool in paracoccidioidomycosis. STUDY DESIGN: Cytologic smears and incisional biopsies were obtained from 10 patients with a clinical suspicion and oral manifestations of paracoccidioidomycosis. Cytologic smears and sections of the incisional biopsy underwent methenamine silver staining for fungi according to the Gomori-Grocott method. The dry glass slides were examined at 400 or 1,000 x magnification, and the presence and shape of yeasts of Paracoccidioides brasiliensis were investigated. RESULTS: Yeasts of the fungus P brasiliensis were clearly identified in cytologic smears and sections from incisional biopsies in all cases analyzed (100.0%). CONCLUSION: Cytology of oral samples proved an effective diagnostic method for the detection of paracoccidioidomycosis in humans. PMID- 11393068 TI - Fixation techniques for fine needle aspiration biopsy smears prepared off site. AB - OBJECTIVE: To identify a simple, cost-effective, reliable fixation method for fine needle aspiration biopsy (FNAB) yielding a specimen suitable for mail transport. STUDY DESIGN: Smears prepared from 59 FNABs of surgical specimens were fixed by continuous fixation in 95% ethanol, spray fixation, air drying, ethanol fixation for either 5 minutes or 4 hours followed by spray fixation, or fixation in 95% ethanol for either 30 minutes or 4 hours followed by air drying. Fixation was graded as unsatisfactory, suboptimal, average, good or excellent. RESULTS: Of smears continuously fixed in ethanol, 96.6% were graded as excellent. Of smears fixed in ethanol followed by spray fixation, 93.2% were excellent irrespective of fixation time; 64.4% of spray-fixed smears were excellent and 27.1% good. Of air dried smears, 93.2% were unsatisfactory or suboptimal; 83.0% of smears fixed in ethanol for 30 minutes and 74.6% of smears fixed for 4 hours prior to air drying were unsatisfactory or suboptimal. CONCLUSION: Fixation of smears in 95% ethanol followed by spray fixation produces excellent results, comparable to those with continuous fixation in ethanol. Spray fixation is generally good but not consistently excellent. Air drying or fixation in ethanol followed by air drying yields unsatisfactory or suboptimal results in most cases. PMID- 11393069 TI - Scratch cytologic findings on surgically resected solitary fibrous tumors of the pleura. AB - OBJECTIVE: To ascertain the cytologic characteristics of solitary fibrous tumors of the pleura (SFTPs) on smear preparations. STUDY DESIGN: Fine needle aspiration cytology (FNAC) was initially attempted preoperatively in five cases, but the specimens were inappropriate for interpretation because only a few tumor cells were obtained. Therefore, scratch smears made at the time of operation were used. Papanicolaou and immunocytochemical staining was performed in all 10 cases, 2 of which were malignant. RESULTS: As expected, cellular tumors yielded more cells. The cytologic appearance was variable, showing spindle/bipolar, dendritic/stellate and intermediate cells. Atypical cells reminiscent of sarcoma were also present in cellular, benign tumors. Highly atypical epithelioid cells were obtained in two malignant cases. Immunocytochemically, the tumor cells were positive for CD34 and vimentin and negative for cytokeratin, regardless of histologic differences and cell shape. CONCLUSION: It seems difficult to diagnose SFTPs with certainty by FNAC, partly because the cell morphology of SFTPs resembles a wide variety of heterogeneous groups of spindle cell tumors and partly because only a few tumor cells were available in the FNAC specimens in the present study. However, a cytologic diagnosis of SFTP is possible if cytologic preparations yield CD34-positive cells with spindle/bipolar or dendritic/stellate morphology. PMID- 11393070 TI - Fine needle aspiration of breast myofibroblastoma. A case report. AB - BACKGROUND: The use of fine needle aspiration cytology (FNAC) for the diagnosis of breast diseases in men has received little attention. We report the cytologic and histologic findings of myofibroblastoma of the breast in a 52-year-old man. CASE: Smears disclosed irregular and cohesive sheets of cells, with ill-defined cytoplasm and oval nuclei containing single nucleoli. The nuclear membrane was frequently grooved, and occasional intranuclear cytoplasmic inclusions (pseudoinclusions) were also found. The background was clean and contained scarce collagenous stroma and fragments of myxoid material. To the best of our knowledge, there have been only seven previous reports of breast myofibroblastoma in which the cytologic features are well documented, and none of them mention the presence of pseudoinclusions. CONCLUSION: FNAC could suggest the diagnosis of this distinctly uncommon tumor if evaluated together with the clinical and radiologic findings. PMID- 11393071 TI - Cytologic diagnosis of peripheral T-cell lymphoma manifesting as ascites. A case report. AB - BACKGROUND: Patients with malignant lymphoma seldom present with effusions without a known history of malignancy. Because of this, initial diagnosis of malignant lymphoma by effusion cytology is uncommon, with few reported cases. CASE: A 75-year-old male presented with fatigue, decreased appetite and progressively increasing abdominal girth over five weeks. Cytologic examination of ascitic fluid obtained by paracentesis revealed non-Hodgkin's lymphoma with a T-cell phenotype, confirmed by immunophenotypic and molecular studies. Approximately one week later, histologic examination of liver and bone marrow revealed involvement by lymphoma, demonstrating immunophenotypic and molecular profiles identical to those obtained from neoplastic lymphocytes recovered from the ascites fluid. CONCLUSION: This case demonstrates a rare presentation of peripheral T-cell lymphoma, clinically manifesting as ascites. In cases such as ours, where the effusion consists predominantly of small to intermediate-sized lymphocytes, distinguishing lymphoma from reactive lymphocytosis may be difficult. This case not only demonstrates the value of effusion cytology for lymphoma diagnosis but also illustrates how the use of various immunophenotypic and molecular techniques may assist the pathologist in properly diagnosing these difficult cases. PMID- 11393072 TI - Fine needle aspiration cytology of a skeletal metastasis of adult Wilms' tumor. A case report. AB - BACKGROUND: Nephroblastoma (Wilms' tumor) is the most common malignant tumor of the kidney in children but is rare in adults. The stage and histopathology of the tumor are the most important prognostic indicators. The common sites of metastasis are lung, liver and lymph nodes. Skeletal metastasis is exceedingly rare in both pediatric and adult nephroblastoma. We report an unusual case of a skeletal metastasis of adult nephroblastoma that developed nine years after the diagnosis of a typical nephroblastoma of favorable histology and that was diagnosed by computed tomography (CT)-guided fine needle aspiration cytology. CASE: Following a right radical nephrectomy for adult nephroblastoma and two local recurrences two and three years later, a 74-year-old woman presented with low back pain. CT and magnetic resonance imaging revealed lytic lesions in the 10th and 12th thoracic vertebrae. Smears prepared from specimens obtained through CT-guided fine needle aspiration biopsy were moderately cellular, with small, round cells arranged singly and in loosely cohesive clusters. These cells had inconspicuous nucleoli and scanty to moderate amounts of cytoplasm. The cells were also positive for cytokeratin and vimentin and appeared similar to areas of blastema in the original tumor. CONCLUSION: A definitive diagnosis of metastatic adult nephroblastoma in thoracic vertebrae was made possible by CT-guided fine needle aspiration cytology in conjunction with clinical and radiologic findings and by using ancillary modalities, such as immunohistochemical studies. PMID- 11393073 TI - Aspiration biopsy findings in amyloid tumor of the cervical vertebra. A case report. AB - BACKGROUND: The differential diagnosis of destructive lytic lesions of the spine includes amyloid tumors. The diagnosis of amyloid tumor with fine needle aspiration biopsy (FNA) is challenging. Previous reports of FNA of osseous amyloid tumors have detailed the cytologic appearance of amyloid along with lymphocytes, plasma cells and histiocytes, occasionally multinucleate or forming granulomatous lesions. CASE: An 84-year-old man presented with neck pain. Radiologic studies showed a destructive, lytic lesion of C-6, with a large, soft tissue mass. FNA yielded many acellular smears containing abundant amyloid that was confirmed with special stains of corresponding tissue cores and subsequent surgical biopsies. CONCLUSION: Osseous amyloid tumors are destructive, lytic lesions that mimic other processes. Amyloid can be distinguished from other substances in FNA samples and amyloid tumor identified, even when amyloid is present without typical cellular components. PMID- 11393074 TI - Malakoplakia of bone. A case report. AB - BACKGROUND: Malakoplakia is an uncommon but distinctive granulomatous disease, characterized by an accumulation of histiocytes or Von Hansemann cells containing intracytoplasmic, laminated Michaelis-Gutmann bodies. CASE: A 3-year-old male presented with a tender swelling in the left gluteal region that had been present for one month. Smears made from a fine needle aspirate showed large histiocytic cells containing intracytoplasmic, basophilic, laminated, targetoid Michaelis Gutmann bodies resembling Von Hansemann cells in malakoplakia. Histopathology confirmed the diagnosis of malakoplakia of bone. CONCLUSION: This case, histologically proven to be malakoplakia, demonstrated regression of the lesion following therapy. The characteristic cytologic features and presence of Von Hansemann cells may in themselves be diagnostic and obviate the need for biopsy. PMID- 11393075 TI - Glassy cell carcinoma of the uterine cervix. Report of a case with cytohistologic and immunohistochemical study. AB - BACKGROUND: Glassy cell carcinomas of the uterine cervix are poorly differentiated carcinomas composed of cells with a large, round to oval nucleus containing one or multiple prominent nucleoli, finely vacuolated eosinophilic to amphophilic cytoplasm and distinct cell borders. These cells occur in sheets and chords, with fibrovascular septae presenting a mixed inflammatory infiltrate. This neoplasm has a poor response to radiotherapy and a worse prognosis than the usual types of adenocarcinoma and squamous cell carcinoma. There are few reports on the cytologic and histopathologic features of this neoplasm. CASE: A 56-year old woman presented with a large, exophytic cervical tumor. Exfoliative cytology showed clusters of cells and single cells with large, round to oval nuclei, with one or multiple nucleoli and moderate to large, finely granulated cytoplasm with distinct cell borders. The background of the smears had a polymorphous inflammatory infiltrate, necrotic debris and proteinaceous material. A high mitotic rate was observed, as were rare bizarre and atypical multinucleated cells. There was no evidence of koilocytes. These findings were highly suggestive of glassy cell carcinoma and were confirmed by the histologic and immunocytochemical findings, with positivity for cytokeratin (MNF116), vimentin and carcinoembryonic antigen and negativity for HMB-45. CONCLUSION: Glassy cell carcinoma of the cervix presents a cytologic picture that can be highly suggestive of the diagnosis in typical cases; however, in difficult cases ancillary techniques, such as immunocytochemistry, as well as histologic findings might confirm the diagnosis. PMID- 11393076 TI - Pulmonary mucormycosis diagnosed by fine needle aspiration cytology. A case report. AB - BACKGROUND: The usefulness of fine needle aspiration cytology (FNAC) in the diagnosis of lung lesions is well documented. Fungal lesions are among nonneoplastic lesions of the lung in which FNAC has proven a useful technique in both immunocompromised and immunocompetent patients. These include cryptococcosis, aspergillosis, histoplasmosis and coccidiodomycosis. Pulmonary mucormycosis, an aggressive fungal infection, is rarely diagnosed on FNAC. We report a case of isolated pulmonary mucormycosis diagnosed on FNAC. CASE: A 62 year-old renal transplant recipient with diabetes mellitus and hypertension, asymptomatic for four months, presented with tachypnea, generalized malaise and weakness. Radiologic studies showed an enlarging, cavitating lesion in the right lung. Computed tomography-guided fine needle aspiration performed on the lung lesion showed fungal profiles with broad, ribbonlike, aseptate hyphae with right angled branching consistent with the Zygomycetes class of fungi, which includes Rhizopus and Mucor species. Fungal cultures confirmed the presence of Rhizopus. The patient underwent right pneumonectomy, was placed on liposomal amphotericin B therapy and discharged with good pulmonary status and stable kidney function. CONCLUSION: FNAC is a useful technique in the diagnosis of pulmonary mucormycosis. PMID- 11393077 TI - Fine needle aspiration cytology of a pancreatic cyst in a patient with autosomal dominant polycystic kidney disease. A case report. AB - BACKGROUND: Solitary cysts occur in approximately 10% of patients with autosomal dominant polycystic kidney disease (ADPKD), and fine needle aspiration cytology (FNAC) can be beneficial in evaluating complications related to the cysts and in excluding malignancies and other cystic lesions that can occur in these patients. CASE: FNAC was performed on a benign epithelial cyst in a symptomatic, 25-year old, white female with ADPKD. The aspirate consisted of scattered small, flat groups of uniform epithelial cells arranged in a honey-comb fashion. CONCLUSION: To the best of our knowledge, this is the first FNAC report of a pancreatic cyst in ADPKD. PMID- 11393078 TI - AIDS-related body cavity-based lymphoma. A case report. AB - BACKGROUND: Body cavity-based lymphomas are rare malignancies in human immunodeficiency virus (HIV)-infected patients, but because of their unusual clinical, morphologic and immunophenotypic features, they are recognized as a distinct subgroup of lymphomas connected to human herpesvirus 8 (HHV-8) infection. CASE: A 39-year-old, HIV-positive, homosexual man was admitted to the hospital because of a left-sided pleural effusion that contained malignant lymphoid cells. He responded partially to a low-dose cyclophosphamide/doxorubycin/vincristine/prednisone regimen and died five months after the diagnosis of lymphoma. On cytology, the sediments contained exclusively large, round, neoplastic, lymphoid cells with abundant basophilic cytoplasm and large, round nuclei with prominent nucleoli. Many cells had immunoblastic features, and some had plasmocytoid differentiation. Mitotic figures were numerous. On flow cytometry, the homogeneous population of large cells expressed CD45, CD38, HLA-DR and CD7 positivity. Other specific T-, B- and NK-cell markers tested negative. Polymerase chain reaction demonstrated Epstein-Barr virus (EBV) and HHV-8 in the malignant effusion. CONCLUSION: Primary effusion from lymphoma with molecular evidence of HHV-8 and EBV coinfection represents a distinct clinical and morphologic entity in AIDS patients. However, immunophenotypic markers of malignant clones can be diverse in different cases. PMID- 11393079 TI - Pelvic abscess from enterobius vermicularis. Report of a case with cytologic detection of eggs and worms. AB - BACKGROUND: Enterobius vermicularis is known to produce perianal and ischioanal abscesses and invade the peritoneal cavity via the female reproductive system, causing pelvic peritonitis. However, there are only rare case reports on the cytodiagnosis of these parasitic lesions. CASE: A 28-year-old woman was admitted with a tender left iliac fossa mass and greenish vaginal discharge. Ultrasonogram and computed tomography scan confirmed the presence of a mass lesion suggestive of a tuboovarian abscess. Cytologic examination of the pus obtained during left salpingo-oophorectomy revealed the presence of ova of E vermicularis and fragments of the adult worm in an inflammatory exudate consisting predominantly of neutrophils, eosinophils and occasional epithelioid cell granulomas. Paraffin sections of the tuboovarian mass showed necrotizing epithelioid cell granulomas, but neither ova nor any worm section was identified. Although the possibility of tuberculosis was considered histologically, Ziehl-Neelsen (Z-N) stain for acid fast bacilli was negative. Z-N staining of the smear and mycobacterial culture of the pus also did not yield positive results. CONCLUSION: E vermicularis may cause tuboovarian abscess with necrotizing epithelioid granulomas mimicking tuberculosis. Cytologic examination of the pus is helpful in the diagnosis. PMID- 11393080 TI - Imprint cytologic features of intracytoplasmic lumina in ependymoma. A report of two cases. AB - BACKGROUND: Intracytoplasmic lumina have been recently recognized as a characteristic histologic feature of ependymoma. However, the cytologic diagnostic usefulness has not been discussed. We encountered two imprint cytology cases of spinal cord ependymomas in which there were intracytoplasmic lumina in the tumor cells. CASES: Two women had spinal cord tumors on magnetic resonance imaging. Imprint cytology study was carried out on the resected tumors. The cytologic specimen of the first case, aged 52, showed tumor clusters consisting of elongated epithelioid cells, a few of which also had intracytoplasmic lumina. Histologically, tumor cells formed ependymal rosettes and pseudoperivascular rosettes. There were a few tumor cells with intracytoplasmic lumina. The cytologic specimen of the second patient, aged 37, had scattered and isolated tumor cells with intracytoplasmic lumina resembling signet-ring cells and paired tumor cells forming small, glandlike structures. Histologically, the tumor was composed mainly of signet-ring-like cells containing intracytoplasmic lumina. CONCLUSION: Intracytoplasmic lumina were observed in the imprint cytologic specimens of spinal cord ependymoma. The diagnosis of ependymomas can be made cytologically when intracytoplasmic lumina are found since no other primary neuroepithelial tumors of the central nervous system possess such a characteristic feature. PMID- 11393081 TI - Trichoblastoma of the skin occurring in the breast. A case report. AB - BACKGROUND: Trichoblastoma is a rare benign skin appendage tumor constituted mostly of follicular germinative cells. It can arise on any part of the body except the palms, soles, nail units and mucosal membranes. No case of it in breast skin has been reported before. Furthermore, fine needle aspiration cytology findings on this lesion have not been described before. CASE: A 76-year old female presented with a firm nodule in her left breast. The tumor was well demarcated, about 1.5 cm in diameter. Fine needle aspiration cytology revealed clusters composed of relatively uniform cells with a high nuclear/cytoplasmic ratio. In the midst of some clusters, the tumor cells had a more abundant cytoplasm. Fibrocellular interstitium or dense cyanophilic acellular material occasionally was attached to them. The tumor cells had oval or fusiform nuclei that had fine, evenly dispersed chromatin. To exclude a diagnosis of breast cancer, it is important to recognize that the clusters are composed of basaloid cells with focal squamous eddies and that there is at least focally peripheral palisading. The histopathologic diagnosis was trichoblastoma. CONCLUSION: Fine needle aspiration cytology can distinguish trichoblastoma from malignant diseases of the breast and may be used to diagnose the lesion in conjunction with clinical findings. PMID- 11393082 TI - Curschmann's spirals in cyst fluid associated with a teratoma of the ovary. A case report. AB - BACKGROUND: Curschmann's spirals, first described more than 100 years ago, are common in cytologic specimens from the respiratory tract and have also been observed in cervical smears, urine, and peritoneal and pleural effusions. No simple theory can explain the exact mode of spiral formation, which is presumed to be a complex physical and biochemical phenomenon. CASE: A 29-year-old woman, gravida 2, para 2, underwent right ovarian cyst excision after an abdominal ultrasound examination revealed a cystic mass of the right ovary. Histologically, the cyst was an immature cystic teratoma containing respiratory epithelium with mucous glands in the submucosa. Cytologic examination of the cyst fluid showed the presence of Curschmann's spirals. CONCLUSION: To the best of our knowledge, the formation of Curschmann's spirals associated with an immature cystic teratoma has not been previously reported. In this situation, the presence of spirals could be explained by formation from mucus normally found in that environment. PMID- 11393083 TI - Fine needle aspiration cytology of late-stage callus in stress fracture. A case report. AB - BACKGROUND: Fine needle aspiration cytology (FNAC) is effective in the diagnosis of bone lesions when combined with careful radiologic and clinical evaluation. The cytologic features of callus have not been described before in the English language literature. CASE: An 18-year-old female presented with a pain in the right lower leg that had been present for two months. Clinical and radiologic findings suggested either stress fracture or periosteal osteosarcoma. The aspiration specimen showed individually scattered, oval cells with moderate amounts of pale pink cytoplasm. The cells contained a single eccentrically located nucleus with evenly distributed, fine chromatin. Osteoclastic giant cells were scattered in the smears. A cytologic diagnosis of benign bone-forming lesion, compatible with callus in fracture, was made. The diagnosis of late-stage callus was confirmed by subsequent histologic examination. CONCLUSION: Typical cases of stress fracture do not need histologic examination, but some cases may be confused with benign and malignant bone tumors. The typical and unique cytologic features of late-stage callus combined with clinical and radiologic findings may prevent the use of more invasive diagnostic procedures and can be a choice for management. PMID- 11393084 TI - Extramedullary plasmacytoma of the parotid gland. Report of a case with extensive amyloid deposition masking the cytologic and histopathologic picture. AB - BACKGROUND: Extramedullary plasmacytomas are uncommon. Although approximately 90% occur in the head and neck region, parotid gland localization is extremely rare. CASE: This report describes fine needle aspiration and histopathologic findings in an extramedullary plasmacytoma arising in the left parotid gland of a 62-year old man. Aspiration smears showed multiple amorphous clumps of material admixed with ductal epithelial cells, multinucleated giant cells and inflammatory cells rich in plasma cells, suggestive of pleomorphic adenoma. In surgical material, excessive amyloid deposition was observed. Six months later the tumor recurred, and in the second surgical specimen clusters of atypical plasma cells among amyloid deposits was noted. Clinical and laboratory examination excluded multiple myeloma. After local recurrence, radiotherapy was applied. Ten months later the patient was well and without systemic involvement. CONCLUSION: Two points are important. First, in the salivary gland region, because of the focal metachromasia of amyloid with Giemsa stain, amyloid can be confused with the chondromyxoid matrix of pleomorphic adenoma. Second, although localized forms of amyloid tumor exist, one should keep in mind that amyloid may be so abundant that it may mask an underlying plasma cell neoplasm, as in our case. PMID- 11393085 TI - Macrophage-rich reaction in lymph nodes as a mimic of metastatic renal cell carcinoma on fine needle aspiration. A case report. AB - BACKGROUND: Renal cell carcinomas have a high metastatic potential. Many of them are occult at initial presentation and mimic a primary neoplasm of the metastatic site. However, not all lymph node enlargements in a patient with a history of renal cell carcinoma are due to metastasis. Foamy macrophages can mimic metastatic renal cell carcinoma cells. CASE: A 60-year-old male with a known diagnosis of renal cell carcinoma of clear cell type developed enlarged neck nodes 44 months after the diagnosis. These were aspirated to yield cystic fluid that, on smears, showed numerous clear cells with low nuclear grade. Immunohistochemical stains revealed these cells to be foamy macrophages (CD68 immunoreactive) and not metastatic renal cell carcinoma, as had been suspected on initial examination of Diff-Quik and Papanicolaou-stained smears. CONCLUSION: Immunohistochemistry is a valuable adjunct in avoiding a false diagnosis of metastatic carcinoma in macrophage-rich nodal reactions in patients with a history of renal cell carcinoma. PMID- 11393086 TI - Fine needle aspiration cytology of primary pulmonary paraganglioma. A case report. AB - BACKGROUND: Primary pulmonary paragangliomas are rare tumors. To our knowledge, there is no prior report on fine needle aspiration cytology (FNAC) in pulmonary paraganglioma. CASE: A 34-year-old man presented with an incidentally found solitary pulmonary mass. FNAC showed papillarylike clusters of epithelioid cells with round to oval nuclei, evenly dispersed chromatin, micronucleoli and occasional anisonucleosis. These cytologic features were suggestive of a sclerosing hemangioma or bronchioloalveolar carcinoma. A right lower lobectomy revealed a primary pulmonary paraganglioma. CONCLUSION: The possibility of pulmonary paraganglioma should be considered in the differential diagnosis of FNAC showing pseudopapillary clusters of epithelioid cells. PMID- 11393087 TI - Cytologic diagnosis of vaginal papillary squamotransitional cell carcinoma. A case report. AB - BACKGROUND: Papillary squamous and squamotransitional cell carcinomas of the cervix and vagina are infrequent morphologic variants of squamous cell carcinoma that may be underdiagnosed due to a bland histologic appearance. To our knowledge, this entity has not been previously detected by Pap smear evaluation. CASE: Vaginal wall pap smears were collected from a patient with a previous hysterectomy for microinvasive cervicovaginal squamous cell carcinoma and extensive carcinoma in situ. The smears were characterized by: (1) large, darkly staining, three-dimensional, branching, papillary epithelial fragments with prominent fibrovascular cores and lined with loosely cohesive epithelial cells; (2) a highly cellular background population of dissociated single epithelial cells with features of severe dysplasia, including hyperchromatic, coarse chromatin; scant, delicate, frayed cytoplasm and karyorrhectic debris; (3) syncytial aggregates of severely dysplastic epithelial cells morphologically similar to the single cells; and (4) lack of a recognizable, morphologically distinct "transitional cell" population. CONCLUSION: Papillary squamotransitional cell carcinoma of the vagina is a rare morphologic variant of squamous cell carcinoma that should be distinguished from benign vaginal squamous papillomas, condylomatous lesions and verrucous carcinoma. However, this lesion is also related to human papillomavirus infection, particularly the high-risk types. Papillary squamotransitional cell carcinoma can be suspected on Pap smear when high grade squamous intraepithelial lesion features are found in combination with three-dimensional papillary tissue fragments with prominent fibrovascular cores. PMID- 11393088 TI - Role of cytomorphology in the evaluation of cutaneous metastases. PMID- 11393089 TI - Fine needle aspiration cytology of soft tissue calcinosis presenting as an enlarging mass in the neck. PMID- 11393090 TI - Fine needle aspiration cytology of well-differentiated fetal adenocarcinoma (endodermal tumor) of the lung. PMID- 11393091 TI - Soft tissue plasmacytoma diagnosed by fine needle aspiration cytology. PMID- 11393092 TI - Fine needle aspiration cytology of macrofollicular variant of thyroid papillary carcinoma in a male. PMID- 11393093 TI - Targeting cytolytic samples for quality control. PMID- 11393094 TI - Interobserver reproducibility: a new approach to quality control by using digital images (D.I. Test). PMID- 11393095 TI - Cytologic diagnosis of an ovarian adenocarcinoma metastatic to the cervix in cervicovaginal smears. PMID- 11393096 TI - Influence of gamma irradiation and storage on apheresis platelets. AB - BACKGROUND AND PURPOSE: Gamma irradiation of platelet concentrates to prevent graft-versus-host disease may inactivate contaminated lymphocytes and subsequently inhibit the synthesis of cytokines in the apheresis platelets during storage. We investigated the influence of irradiation and storage on apheresis platelets collected with the COBE Spectra or Fenwal CS-3000 Plus systems. METHODS: Eleven units of apheresis platelets were collected with a COBE Spectra cell separator and another 11 units with a Fenwal CS-3000 Plus system. Each unit of apheresis platelets was divided into two equal parts: one was irradiated with 3000 cGy directly after blood donation, and the other served as a control. Cell counts, platelet activation marker CD62 antigen, blood gas values, and supernatant concentrations of K+, Na+, lactate, glucose, interleukin-1 beta (IL-1 beta), IL-8, and tumor necrosis factor-alpha (TNF-alpha) were determined in paired samples on the day of collection (day 0) and after 5 days of storage (day 5). RESULTS: No significant differences in white cell counts or TNF-alpha concentrations were noted between the irradiated and control platelets on day 0 or day 5, whereas the mean proportion of platelets expressing CD62P (22.65% vs 25%, p = 0.014) and the mean IL-1 beta (45.55 pg/mL vs 52.75 pg/mL, p = 0.004) and IL-8 concentrations (10.68 pg/mL vs 13.07 pg/mL, p = 0.015) were significantly lower in irradiated than control platelets on day 5. The 5-day storage significantly increased the mean proportion of platelets expressing CD62P (25.00% vs 15.02%, p = 0.008), mean PO2 (116.34 mm Hg vs 98.07 mm Hg, p = 0.002), and mean concentrations of K+ (3.30 mmol/L vs 3.06 mmol/L, p < 0.001), lactate (15.12 mmol/L vs 3.23 mmol/L, p < 0.001), IL-1 beta (52.75 pg/mL vs 29.73 pg/mL, p = 0.001), and IL-8 (13.07 pg/mL vs 3.62 pg/mL, p < 0.001). Five-day storage also significantly decreased white cell count (0.18 x 10(8) vs 0.74 x 10(8), p < 0.001), PCO2 (19.38 mm Hg vs 50.51 mm Hg, p < 0.001), and concentrations of HCO3- (10.36 mmol/L vs 21.34 mmol/L, p < 0.001) and glucose (193.37 mg/dL vs 309.18 mg/dL, p < 0.001). Platelet counts and concentrations of IL-1 beta, IL-8, and TNF alpha on day 0 did not differ significantly between control apheresis platelets collected with the Fenwal CS-3000 Plus and those collected with COBE Spectra. The mean white cell count (1.29 x 10(8) vs 0.19 x 10(8), p = 0.002) and the proportion of platelets expressing CD62P (24.71% vs 7.09%, p < 0.001) on day 0, however, were significantly higher in the platelets collected with the Fenwal CS3000-Plus than in those collected with the COBE Spectra. CONCLUSIONS: Gamma irradiation of apheresis platelets inhibits the expression of platelet CD62P and the secretion of IL-1 beta and IL-8 after 5 days' storage. PMID- 11393097 TI - Ultrasonographic and cytologic findings of metastatic cancer in the thyroid gland. AB - BACKGROUND AND PURPOSE: Cancer metastasizing to the thyroid is not uncommon. Prompt diagnosis and aggressive treatment appear to contribute to better prognosis in some patients. The aim of this study was to determine the clinical manifestations, ultrasonographic and cytologic findings, and outcomes in patients with metastatic thyroid cancer. METHODS: We retrospectively reviewed the medical records, including clinical courses, ultrasonographic and cytologic findings, and outcomes, of 14 patients with metastatic thyroid cancer. RESULTS: There were various primary sites of cancer in our study population, with adenocarcinoma the most common. The sonographic images in five of 12 patients who underwent sonography showed diffusely heterogeneous hypoechoic abnormalities, while the remaining seven patients had nodular lesions. The cytologic appearances of metastatic lesions in the thyroid were characteristic of the primary malignancies. CONCLUSIONS: Most cancer metastatic to the thyroid presented as advanced metastases of primary malignancies. The sonographic appearance alone was not the ideal discriminator, and fine needle aspiration cytology helped define the diagnosis and management strategy. PMID- 11393098 TI - Effect of percent free prostate-specific antigen measurement on improving the specificity of serum prostate-specific antigen testing in Taiwanese patients. AB - BACKGROUND AND PURPOSE: Measurement of percent free prostate-specific antigen (fPSA) has been shown to enhance the diagnostic performance of traditional serum PSA determination. We sought to determine whether measurement of percent fPSA could improve the specificity of serum total PSA testing in the detection of prostate cancer in Taiwanese patients with intermediate serum PSA concentrations. METHODS: The medical records of 211 patients examined from March 1998 through March 2000 were analyzed retrospectively; all had a serum total PSA concentration of between 4.1 and 20.0 ng/mL and negative digital rectal examination (DRE) findings, and had undergone a prostate biopsy. Biopsy results were correlated with the serum total PSA concentration and percent fPSA, which were determined using a microparticle enzyme immunoassay. Percent fPSA was calculated as the ratio of fPSA to total PSA multiple by 100. The sensitivity and specificity were calculated and the receiver operating characteristic (ROC) curves were generated from different cutoff values of percent fPSA and total PSA. RESULTS: Thirty-four patients (16.1%) had positive biopsy results (prostate cancer). Patients with positive biopsy results had significantly lower percent fPSA values than those with negative results (11.7% vs 16.0%, p < 0.001). Patients with a lower percent fPSA (< 10%) had a higher probability of a positive biopsy result than those with a high percent fPSA (> 20%) (positive biopsy rate, 29% vs 10%, p < 0.05). Using a cutoff value of 25% fPSA or below, the sensitivity, specificity, and positive predictive value in differentiating patients with positive biopsy results from those with negative results were 97%, 13%, and 18%, respectively. Consequently, 27 unnecessary biopsies could have been avoided at the cost of missing one cancer. The area under the ROC curve was 0.68 for percent fPSA and 0.63 for total PSA (p > 0.05). CONCLUSIONS: For Taiwanese patients with a serum PSA concentration of between 4.1 and 20.0 ng/mL, the incidence of prostate cancer is relatively low; measurement of percent fPSA only weakly enhances the specificity of serum PSA testing in detecting prostate cancer. PMID- 11393099 TI - Effectiveness of a multidisciplinary rehabilitation program in elderly patients with hip fractures. AB - BACKGROUND AND PURPOSE: Hip fracture is a significant health challenge to the elderly, with a high risk of complications, long hospital stay, and functional deterioration. The purpose of this study was to evaluate the effectiveness of a multidisciplinary rehabilitation program (MRP) in older patients with hip fractures. METHODS: A before-and-after quasi-experimental design was used. Data were collected at a large teaching hospital in Taipei. The first 105 consecutive patients admitted received conventional care (control group). The next 50 consecutive patients received physical therapy, nurse-supervised exercise, and discharge planning (intervention group). Subjects were assessed upon admission, on the fourth postoperative day, and on the discharge day. The 155 patients recruited met the following criteria: 60 years of age or older, and hospitalized to receive either closed reduction and internal fixation, or hip arthroplasty. RESULTS: Compared with the control group, the intervention group had a shorter average hospital stay (9.9 vs 11.6 days, p = 0.01), earlier ambulation (5.7 vs 6.5 days postoperatively), and a lower rate of bowel incontinence (4.0% vs 23.8%) and pressure sores (14.0% vs 24.8%) during hospitalization. The intervention group experienced less decline in mobility (relative to pre-fracture status) than the control group. No significant difference was found in other outcome variables between the control and intervention groups. CONCLUSIONS: The findings of this study indicate that an MRP involving physical therapy, nurse-supervised exercise, and discharge planning positively affects health outcomes in older patients with hip fractures. PMID- 11393100 TI - Langerhans' cell histiocytosis presenting with a para-aortic lesion and heart failure. AB - Langerhans' cell histiocytosis (LCH) is an uncommon disease with variable manifestations. We report a case of LCH with the unusual initial presentations of chest pain and progressive heart failure in a 5-year-old boy. Chest radiography revealed a wide mediastinum with cardiomegaly. Electrocardiography showed first degree atrioventricular block and an inverted T wave over V4-V6. Echocardiography, computed tomography, and magnetic resonance imaging of the chest all showed an infiltrating lesion that enveloped the entire heart, great vessels, and coronary arteries. Pathologic examination of the biopsy specimen revealed LCH. Chemotherapy, which included prednisolone, vincristine, methotrexate, and 6-mercaptopurine, had only a minimal effect on the tumor. After the addition of etoposide, the lesion decreased in size, and the symptoms and signs of heart failure and chest pain were ameliorated. PMID- 11393101 TI - Perforated peptic ulcer in an infant. AB - We describe a case of perforated peptic ulcer (PPU) in a 9-month-old boy. Abdominal distension was the first clinical sign of PPU. Before he developed abdominal distension, the patient had suffered from an upper respiratory tract infection with fever for about 2 weeks, which was treated intermittently with ibuprofen. A plain abdominal radiograph revealed pneumoperitoneum with a football sign. At laparotomy, a 0.8-cm perforated hole was found over the prepyloric area. Simple closure with omental patching was performed after debridement of the perforation. Pathologic examination showed chronic peptic ulcer with Helicobacter pylori infection. The postoperative course and outcome were satisfactory. The stress of underlying disease, use of ibuprofen, blood type (A), and H. pylori infection might have contributed to the development of PPU in this patient. PPU in infancy is rare and has a high mortality rate; early recognition and prompt surgical intervention are key to successful management. PMID- 11393102 TI - Sigmoid volvulus in children: report of two cases. AB - Volvulus of the sigmoid colon is rare in children. An early, accurate diagnosis can avoid unnecessary surgery and reduce the risk of complications. This condition is mainly due to a redundant sigmoid colon with a narrow mesosigmoid attachment. We describe two cases of sigmoid volvulus, which showed different clinical severities and were treated with different methods. Patient 1, a 9-year old boy, presented with acute abdominal pain and vomiting. Patient 2, an 11-year old boy, presented with abdominal pain, abdominal distention, and bloody mucoid stool. Plain abdominal radiographs revealed a distended colonic loop extending upward from the pelvis in patient 1 and a typical "coffee bean" sign in patient 2. Barium enema examination was used to confirm the diagnosis in both cases. The volvulus was reduced by insertion of a rectal tube in patient 1 and surgically in patient 2. Sigmoid colon volvulus should be included in the differential diagnosis of childhood abdominal pain or distention. This report suggests that nonsurgical reduction should be attempted first for uncompromised sigmoid volvulus in children, unless bowel ischemia or perforation develops. PMID- 11393103 TI - Paget's disease of the bone: a case report. AB - Paget's disease of bone is rare in Asia. We report a case of Paget's disease in a 58-year-old Taiwanese man who was admitted with a 3-month history of bilateral numbness in the buttock region. Laboratory data disclosed an elevated serum alkaline phosphatase level (510 U/L). Plain radiographs of the lumbar spine showed generalized increased density at the third lumbar vertebra, associated with cortical thickening, loss of cortico-cancellous definition, and increased anteroposterior diameter. The T1-weighted magnetic resonance image of the lumbar spine showed diffuse, heterogeneous low signal intensity at the third lumbar vertebral body, pedicle, laminae, and spinal process; these areas showed mixed high and low signal intensity on the T2-weighted image. Technetium-99m bone scan revealed abnormal uptake in the involved vertebra. Histologic examination of the third lumbar spinal process confirmed the diagnosis of Paget's disease of bone. The patient remained well during a follow-up period of 6 months. PMID- 11393104 TI - Pneumocephalus secondary to septic thrombosis of the superior sagittal sinus: report of a case. AB - Pneumocephalus secondary to septic superior sagittal sinus thrombosis (SSSST) is extremely rare. We report computed tomography (CT) findings in a 63-year-old man with SSSST caused by the gas-forming organism Klebsiella pneumoniae. The patient presented with fever, chills, general weakness, and spontaneous progressive swelling of the right frontoparietal scalp. CT revealed a gas-containing abscess over the right frontoparietal subgaleal region and in the superior sagittal sinus. Surgical drainage of the subgaleal abscess was performed and blood and pus cultures grew Klebsiella pneumoniae. The patient died of sepsis on the 6th day of hospitalization. PMID- 11393105 TI - Modified arterial switch operation by sharing the common wall between the great arteries. AB - BACKGROUND: Suprapulmonary stenosis and coronary arterial obstruction still remain as problems after an arterial switch operation (ASO). We used a modified ASO applying the common wall and in situ transfer techniques to improve the current procedure. METHODS: From October 1996 to December 1997, 11 babies aged 6 days to 3 months with transposition of the great arteries underwent a modified ASO which included sharing the common wall between the great arteries until above the anterior neoaortic suture-line for coronary and pulmonary artery reconstruction. Coronary arteries were of usual type in three cases, juxtacommissural origin in five, and a high takeoff in one; all were redirected almost in situ. RESULTS: There was no early death (< 30 d), coronary or bleeding problems. One late death occurred after a repeat surgery for suprapulmonary stenosis. This was caused by upward stretching of the left pulmonary artery, which was placed above the high left-sided neoaortic anastomosis for in situ transfer of the high takeoff coronary arteries. Intraluminal growth of the adventitia also contributed to suprapulmonary stenosis, which decreased significantly when the common wall adventitia was cleaned in the last two cases we operated on. Ten patients were doing well at follow-up (30.9 +/- 5.2 mo). CONCLUSIONS: This modified ASO by common wall and in situ transfer might avoid coronary kinking and lessen the chance of postoperative bleeding. To avoid suprapulmonary stenosis, common wall adventitia inside the pulmonary pathway should be cleaned, and the left and right pulmonary arteries should also be kept in situ as possible as in coronary redirection. PMID- 11393106 TI - Comparison of ischemic patterns in myocardial bridge and syndrome X: evaluation by dobutamine stress echocardiography and stress thallium-201 SPECT. AB - BACKGROUND AND PURPOSE: Ischemic patterns in patients with syndrome X are thought to differ from those in patients with myocardial bridge, because the mechanisms of coronary flow reduction in these two diseases are different. The aim of this study was to compare the ischemic patterns in patients with syndrome X and those with myocardial bridge through the use of dobutamine stress echocardiography (DSE) and stress thallium-201 single-photon emission computed tomography (SPECT). METHODS: Twenty-six patients with typical angina and stress-induced ST-segment depression were enrolled. All patients underwent coronary angiography, DSE, and stress thallium-201 SPECT within 7 days after enrollment. RESULTS: Of the 26 patients enrolled, 10 had myocardial bridge of the left anterior descending artery and 16 had syndrome X. Among patients with myocardial bridge, myocardial dyssynergy was found by DSE in five patients and reversible or fixed thallium-201 perfusion defects were found in four. Seven patients with myocardial bridge had reverse redistribution patterns on thallium-201 scintigraphy. In the 16 patients with syndrome X, myocardial dyssynergy was found by DSE in only one patient (p = 0.018 vs myocardial bridge group) and reversible or fixed thallium-201 perfusion defects were found in nine (p > 0.05 vs myocardial bridge group). Four patients with syndrome X had reverse redistribution patterns on thallium-201 scintigraphy. The resting left ventricular end-diastolic pressure was higher in patients with myocardial bridge than in those with syndrome X (17 +/- 4 vs 12 +/- 5 mm Hg, p = 0.02). CONCLUSIONS: The most common ischemic patterns in patients with syndrome X were chest pain and stress-induced ST-segment depression, followed by myocardial perfusion defects. Dobutamine-induced dyssynergy was rare. Left ventricular end diastolic pressure elevation and dobutamine-induced wall motion abnormalities were more common in patients with myocardial bridge than in those with syndrome X. PMID- 11393107 TI - Subtyping HIV-1 infections in Taiwan using peptide-enzyme immunoassay, reverse transcription-polymerase chain reaction, and sequencing. AB - BACKGROUND AND PURPOSE: There are important questions about epidemiologic transmission patterns as well as the possibility that genetic and phenotypic differences in human immunodeficiency virus type 1 (HIV-1) affect transmissibility, infectivity, pathogenicity, and response to therapy and vaccines. To delinate the genetic heterogeneity of HIV-1 and the association of subtypes with risk factors and location of residence in Taiwan, subtypes of HIV-1 in Taiwanese patients were identified and a phylogenetic study was performed. In addition, the accuracy of peptide-enzyme immunoassay (EIA) using serum samples from Taiwanese patients infected with HIV-1 was investigated. METHODS: Peptide EIA was used to give a preliminary subtype of HIV-1-positive serum samples collected from different areas of Taiwan. Reverse transcription (RT)-polymerase chain reaction (PCR) and genetic sequencing were used to confirm the peptide-EIA results and to construct a phylogenetic tree. RESULTS: Among the 149 serum samples, 98 were subtype B (66%), 38 subtype E (25%), two subtype Thai-B (1.3%), one subtype G (0.7%), and one subtype C (0.7%). Comparison of risk factors for HIV-1 infection and subtype revealed that most B subtype infections (59/98) occurred in homosexual or heterosexual patients, whereas 28 of 38 E subtype infections occurred in heterosexual patients. The B/E ratio was significantly different (p < 0.05) in Taipei than in other areas of Taiwan. CONCLUSIONS: These results suggest that the predominant subtype of HIV-1 infection in Taiwan is B, followed by E, and that the distribution of HIV-1 subtypes in Taiwan is similar to that of Thailand, although the genetic sequences are distinct. Homosexuality, heterosexuality, bisexuality, and intravenous drug use behaviors affect the distribution of different subtypes of HIV-1 infection. Peptide-EIA in conjunction with RT-PCR and sequencing can provide accurate subtyping of HIV-1 infection. PMID- 11393108 TI - Antibiotic use in public hospitals in Taiwan after the implementation of National Health Insurance. AB - BACKGROUND AND PURPOSE: Only limited data is available on the patterns of antibiotic use in hospitals in Taiwan before and after the implementation of national health insurance. This study aimed to determine the patterns of use of antibiotics in public hospitals in Taiwan after the implementation of the National Health Insurance program and to compare these with patterns prior to the implementation. METHODS: Data on the annual use of all antibiotics in public hospitals in Taiwan during the period from fiscal year (FY) 1994-1995 to 1997 1998 were collected and analyzed. Hospitals included seven medical centers, 19 regional hospitals, 53 district hospitals, and eight specialty service hospitals. RESULTS: The annual cost of antibiotics made up 17.4% of the annual cost of all medication used in these hospitals in FY 1994-1995, and decreased year by year to 12.6% in FY 1997-1998. During the study period, 57.4% of the total cost of antibiotics were incurred by medical centers, 24.6% by regional hospitals, 16.2% by district hospitals, and 1.8% by specialty service hospitals. The most commonly used class of antibiotic was cephalosporins, accounting for 48.0% to 54.3% of total antibiotic costs. The second most commonly used class of antibiotic was penicillins, accounting for 15.9% to 17.4% of total antibiotic costs. In FY 1994 1995, the next most commonly used classes of antibiotics were aminoglycosides, fluoroquinolones, and other beta-lactams, respectively, but by FY 1997-1998 these had changed to fluoroquinolones, glycopeptides, and aminoglycosides, respectively. Among the various cephalosporins used, first-generation cephalosporins accounted for 76.1% of all cephalosporins used in FY 1994-1995, which increased year by year to 84.0% in FY 1997-1998. Second-generation cephalosporins accounted for 20.7% of all cephalosporins used in FY 1994-1995, decreasing to 13.2% in FY 1997-1998. CONCLUSION: The introduction of the National Health Insurance program in Taiwan brought about a major change in antibiotic use patterns in public hospitals. PMID- 11393109 TI - Valveless outflow reconstruction using autologous tissue as a posterior wall for pulmonary atresia with ventricular septal defect. AB - BACKGROUND AND PURPOSE: Reoperation is inevitable for some patients with pulmonary atresia who receive a heterograft or homograft in a primary Rastelli operation. Nonetheless, the need for reoperation in patients with classic Fallot's tetralogy who have undergone total correction with a transannular patch is unusual. We sought to change pulmonary atresia into Fallot's tetralogy and used a transannular patch instead of the conventional Rastelli operation. PATIENTS AND METHODS: Valveless outflow direct reconstruction was performed on 10 consecutive patients with pulmonary atresia and ventricular septal defect between August 1997 and 1999. Patient ages ranged from 1.3 to 11.5 years. A Blalock Taussig shunt was previously constructed in four of these patients and a central shunt was constructed in five. The major aortopulmonary collateral arteries were occluded in one patient by repeated coil embolization after the central shunt. The strategy was to connect the right ventriculotomy with the pulmonary arteries directly, even if there was a gap with a long atretic cord. In patients with a previous central shunt covered with a Gore-Tex membrane, the reactive visceral pericardium over the in situ tissue (the left atrium, right ventricle, or aorta) was used as the autologous posterior wall. Thus, only autologous, fresh pericardium without a valve was used to cover the anterior part of the right ventricular outflow tract, as in the repair of classic Fallot's tetralogy with a transannular patch. RESULTS: There was no mortality, and the postoperative central venous pressure was low in all patients. No gradient was noted across the right ventricular outflow tract. Follow-up echocardiography revealed a competent tricuspid valve with mild pulmonary regurgitation in all patients. CONCLUSIONS: The results of this study suggest that valveless outflow direct reconstruction provides adequate pulmonary circulation without hypertension in pulmonary atresia patients with a ventricular septal defect if the tricuspid valve is competent. PMID- 11393110 TI - Risk factors for low birth weight among first-time mothers in southern Taiwan. AB - BACKGROUND AND PURPOSE: Low birth weight (LBW) greatly affects infant development, the family, and health care financial systems. Nonetheless, the risk factors for LBW in Taiwan have not been investigated. The purpose of this study was to determine the risk factors for LBW in Kaohsiung County. METHODS: In this cross-sectional study, we recruited 1,147 first-time mothers, including all adolescent mothers (< 20 years old) who gave birth during the period from June 1994 through May 1995 and all adult mothers (> or = 20 years old) who gave birth in January and February 1995. The subjects were interviewed during home visits or by telephone by public health nurses in each township. Logistic regression analysis was used to identify the risk factors for LBW in adolescent and adult mothers. RESULTS: The prevalence of LBW in the study population was 6.2%, ranging from 7.6% in adolescent mothers to 4.9% in adult mothers. Univariate analysis showed that mothers who had low pregravid weight (< 45 kg), infrequent prenatal visits (< 10), anemia, low gestational weight gain (< 10 kg), or habits of alcohol consumption or cigarette smoking were more likely to give birth to LBW infants. In logistic regression analysis, after controlling for covariates, the significant risk factors for LBW were low gestational weight gain (< 10 kg) and low pregravid weight (< 45 kg) for adolescent mothers. Infrequent prenatal visits (< 10) was the only significant risk factor for adult mothers. CONCLUSIONS: In this study, the risk factors for LBW among adolescent and adult mothers were different. This suggests that programs for prevention of LBW should be tailored according to maternal age, begin before conception, and continue with enhanced surveillance during prenatal visits. PMID- 11393111 TI - Prognostic indicators of survival in infants with congenital diaphragmatic hernia. AB - PURPOSE: This study sought to identify the factors predictive of the short-term outcome in infants with congenital diaphragmatic hernia (CDH). METHODS: We retrospectively reviewed medical records from September 1985 to December 1998 for all infants born with CDH and managed at National Taiwan University Hospital (NTUH). Coexisting pathology and measures of respiratory function were analyzed to determine the prognostic factors. RESULTS: A total of 32 infants with CDH were managed at NTUH over the past 13 years. The investigated factors associated with poor prognosis in CDH included congenital heart disease, chromosomal abnormality, pneumothorax, and preoperative parameters including arterial partial carbon dioxide pressure greater than 40 mmHg, arterial partial oxygen pressure less than 100 mmHg, alveolo-arterial partial oxygen pressure greater than 610 mmHg, and oxygen index (OI) greater than 0.4. Multiple logistic regression analyses indicated that only an OI greater than 0.4 and pneumothorax were significant indicators of poor prognosis. When the incidence of pneumothorax was compared between patients who received ventilation using a conventional mechanical ventilator or high-frequency oscillatory ventilator (HFOV), a trend toward a lower incidence of pneumothorax in those using HFOV for initial stabilization was found (p = 0.08). CONCLUSIONS: An OI greater than 0.4 before surgery and pneumothorax are poor prognostic indicators in infants with CDH. A high OI is reflective of the severity of pulmonary dysfunction in infants. Pneumothorax further compromises the dysfunction of a hypoplastic lung in infants with CDH. The results of this study show the importance of avoiding iatrogenic pneumothorax during management of infants with CDH. PMID- 11393112 TI - Comparison of the metabolic effects of metformin and troglitazone on fructose induced insulin resistance in male Sprague-Dawley rats. AB - BACKGROUND AND PURPOSE: Insulin resistance is a hallmark of the development of type 2 diabetes. Metformin and troglitazone are oral antidiabetic agents used to reduce insulin resistance. The aim of this study was to compare the metabolic effects of these two drugs in fructose-induced insulin-resistant rodents. METHODS: Male Sprague-Dawley rats were allocated to receive one of the following four treatments for 6 weeks: normal rat chow (control group, n = 7), high fructose diet (fructose group, n = 7), high-fructose diet plus metformin (metformin group, n = 8), or high-fructose diet plus troglitazone (troglitazone group, n = 8). Systolic blood pressure (SBP), insulin, free fatty acid (FFA), and triglyceride concentrations were measured as parameters of insulin resistance. Leptin concentration was also measured in the four groups. RESULTS: The fructose group developed significantly elevated SBP, hyperinsulinemia, and hypertriglyceridemia without significant change in body weight or leptin concentration compared with the control group. The metformin group had significantly reduced body weight (397.9 +/- 40.9 vs 470.1 +/- 59.6 g, p < 0.05), insulin concentration (14.8 +/- 10.5 vs 48.4 +/- 15.2 microU/mL, p < 0.05), triglyceride concentration (75.3 +/- 65.5 vs 250.1 +/- 95.7 mg/dL, p < 0.05), and leptin concentration (3.1 +/- 1.5 vs 6.9 +/- 2.0 ng/mL, p < 0.05) without significant change in SBP (147.8 +/- 5.8 vs 152.4 +/- 13.0 mm Hg, p > 0.05) compared with the fructose group. The troglitazone group had significantly reduced SBP (137.8 +/- 9.2 vs 152.4 +/- 13.0 mm Hg, p < 0.05), insulin concentration (15.0 +/- 13.6 vs 48.4 +/- 15.2 microU/mL, p < 0.05), FFA concentration (38.9 +/- 22.7 vs 78.7 +/- 24.6 mg/dL, p < 0.05), triglyceride concentration (67.6 +/- 32.4 vs 250.1 +/- 95.7 mg/dL, p < 0.05), and leptin concentration (4.4 +/- 2.0 vs 6.9 +/- 2.0 ng/mL, p < 0.05) without significant change in body weight (452.5 +/- 32.8 vs 470.1 +/- 59.6 g, p > 0.05) compared with the fructose group. The metabolic effects of metformin and troglitazone on insulin, FFA, triglyceride, and leptin concentrations were not significantly different. However, metformin treatment resulted in significantly lower body weight (397.9 +/- 40.9 vs 452.5 +/- 32.8 g) and troglitazone treatment in significantly lower SBP (137.8 +/- 9.2 vs 147.8 +/- 5.8 mm Hg) compared to the fructose group, after adjusting for basal values (p < 0.05). CONCLUSIONS: Both metformin and troglitazone were comparably effective in reducing insulin resistance. Metformin treatment caused body weight reduction but was not effective in reducing SBP. Troglitazone treatment lowered SBP but did not reduce body weight. PMID- 11393113 TI - Synergistic effect of Nd:YAG laser combined with fluoride varnish on inhibition of caries formation in dental pits and fissures in vitro. AB - BACKGROUND AND PURPOSE: Although the effectiveness of neodymium yttrium aluminum garnet (Nd:YAG) laser and fluoride anticaries treatment has been established, most previous studies focused on smooth tooth surfaces. We evaluated the anticaries effects of Nd:YAG laser combined with fluoride varnish (Duraphat) on caries-susceptible pit and fissure areas. METHODS: A total of 36 noncarious molars were treated with either a Nd:YAG laser (2.5 W, 6 sec) followed by fluoride varnish, Nd:YAG laser only, fluoride varnish only, or no treatment (control). Artificial carious lesions were created to assess the acid resistance of enamel after treatment. Undecalcified successive tooth slices were made. Percentage lesion formation, lesion length, and lesion depth were evaluated using polarized light microscopy. RESULTS: The Nd:YAG laser enhanced the resistance of dental enamel to acid challenge. However, Nd:YAG laser alone was not as effective as the Nd:YAG laser combined with fluoride varnish, especially for the treatment of pits and fissures. Nd:YAG laser treatment combined with fluoride varnish inhibited 43% of lesions at pits and fissures and 80% of lesions on smooth surfaces compared to no treatment. Carious lesions had shallower depth and shorter length. No carious lesion extended beyond the dentinoenamel junction in either laser-treatment group. CONCLUSIONS: A synergistic effect on dental caries prevention in pit and fissure areas and on the smooth surfaces of the tooth can be achieved by applying Nd:YAG laser followed by fluoride varnish. PMID- 11393114 TI - Effectiveness of transperitoneal and trans-retroperitoneal laparoscopic adrenalectomy versus open adrenalectomy. AB - PURPOSE: This study compared the effectiveness of laparoscopic adrenalectomy, using either a transperitoneal or trans-retroperitoneal approach, with that of open adrenalectomy in patients with benign adrenal tumors. METHODS: From February 1995 to April 2000, laparoscopic adrenalectomy was performed on 31 patients with adrenal tumors, including 16 aldosteronomas, 10 Cushing's adenomas, three nonfunctioning tumors, and two pheochromocytomas. A lateral trans-retroperitoneal approach was used for the first 16 patients and a lateral transperitoneal approach was used for the last 15 patients. Twenty-one patients who received open adrenalectomy during the same period served as a control group. Comparisons were made between laparoscopy and open groups, and between transperitoneal and trans retroperitoneal groups. RESULTS: Conversion to open adrenalectomy was necessary in two cases--both in the trans-retroperitoneal group during the first 2 years of the study period. No other intraoperative complications occurred and blood transfusion was not used. Compared with the open group, the laparoscopic group had less blood loss (71 vs 124 mL), resumed oral feeding earlier (28 vs 60 hr), required less postoperative narcotics (45 vs 120 mg meperidine), and had shorter postoperative hospital stays (4.9 vs 7.6 days) (all p < 0.05). The mean operative time was longer in the laparoscopic group (203 vs 123 min, p < 0.001). There were no significant differences between the transperitoneal and trans-retroperitoneal laparoscopy groups in any of the studied parameters, except that the operative time was longer in the trans-retroperitoneal laparoscopy group (244 vs 166 minutes, p < 0.01). CONCLUSIONS: Decreased blood loss, less postoperative pain, earlier resumption of oral feeding, and shorter hospital stays were achieved in patients undergoing laparoscopic adrenalectomy. These findings indicate that laparoscopic adrenalectomy is the treatment of choice for benign adrenal tumors. The transperitoneal approach yielded shorter operative time than the trans retroperitoneal approach, because it offered a clearer view and familiar landmarks. PMID- 11393115 TI - Cadaveric study of blood supply to the lower intraorbital fat: etiologic relevance to the complication of anaerobic cellulitis in orbital floor fracture. AB - BACKGROUND AND PURPOSE: Although orbital fractures are common, orbital cellulitis rarely develops following orbital fracture. We hypothesized that compromise of the blood supply to the intraorbital fat during orbital floor fracture is responsible for this condition. The purpose of this study was to determine whether or not the lower intraorbital fat is supplied by a branch of the infraorbital artery along the orbital groove or canal on the orbital floor. MATERIALS AND METHODS: We dissected 14 orbits from seven fixed human cadavers and 12 orbits from six fresh cadaver heads following dye injection into the maxillary artery. The sites of dye-filled vessels branching from the infraorbital artery supplying the lower intraorbital fat were measured and plotted on a two dimensional orbital floor graph. RESULTS: A main branch of the infraorbital artery rose through the medial orbital floor to supply the lower intraorbital fat in all of the cadaver orbits. The sites of the branching point of the vessel ranged from 0 to 5 mm (mean, 2.2 mm; n = 14) medial to the line connecting the infraorbital foramen and the infraorbital groove. The shortest distance measured from the branching point to the orbital rim ranged from 3 to 20 mm (mean, 14.1 mm; n = 14). This suggests that if orbital fracture were to occur around the infraorbital groove or canal, this vascular pedicle would be in danger of being incarcerated by bone fragments. CONCLUSION: Our cadaveric investigation revealed that the lower intraorbital fat is supplied by a branch of the infraorbital artery along the infraorbital groove or canal on the orbital floor. This finding suggests that compromised blood supply to the intraorbital fat may cause anaerobic cellulitis or enophthalmos. PMID- 11393116 TI - Thickened pituitary stalk with central diabetes insipidus: report of three cases. AB - Diabetes insipidus of central origin usually results from lesions in the hypothalamic neurohypophyseal system. Lymphocytic infundibuloneurohypophysitis is an uncommon cause. Cases of lymphocytic infundibuloneurohypophysitis with thickening of the pituitary stalk and enlargement of the neurohypophysis with no hyperintense signal in the posterior pituitary have been reported. Reported cases presenting with isolated thickening of the pituitary stalk are very rare. We report three such cases, one in a nulliparous woman and the other two in men. Magnetic resonance (MR) imaging in these patients revealed isolated thickening of the pituitary stalk, loss of the hyperintense signal of the posterior pituitary, and an adenohypophysis of normal size. All cases had abnormal nodular infundibular enlargement. One male patient had hypogonadism; the other patients showed no sign of adenohypophyseal deficiency on stimulation test. Serial follow up MR imaging revealed that all three patients had persistent thickening of the pituitary stalk. Diabetes insipidus was controlled by the administration of desmopressin acetate in all patients. PMID- 11393117 TI - Acute paraplegia in a patient with spinal tophi: a case report. AB - A 28-year-old man with a 5-year history of gouty arthritis suffered from an acute episode of lower back pain. He visited a rehabilitative clinic and received physical therapy following his examination. Weakness and numbness of both lower legs developed rapidly after physical therapy. He was sent to our hospital with complete paralysis of both lower limbs and complete sensory loss below the umbilicus 3 hours after the physical therapy. No peripheral tophi were found. Myelography showed an extrinsic compression of the dura sac at T10. Emergency decompressive laminectomy of T9 to T11 was performed. During the surgery, caseous material was found deposited in the ligamentum flavum and the left T9 to T10 facet joint, with indentation of the dura sac. The pathologic diagnosis was spinal tophi. After surgery, the patient's neurologic function recovered rapidly. It was suspected that inappropriate physical therapy might have aggravated acute inflammation of spinal gout and resulted in a rapid deterioration of neurologic function. Though gout is a chronic medical disease, an acute attack of spinal gout may be disastrous and requires emergency neurosurgical intervention. PMID- 11393118 TI - Paraganglioma manifesting as shock: a case report. AB - Paraganglioma is a rare neuroendocrine tumor in children that rarely manifests as shock. We describe the case of a 12-year-old girl with paraganglioma who developed impaired cardiac function, pulmonary edema, and shock at the time of admission. Her blood pressure stabilized after intravenous normal saline rescue and dopamine treatment. However, hypertension was noted thereafter. After a series of examinations, paraganglioma was diagnosed and excision of the tumor was performed. After surgery, blood pressure stabilized and her cardiac function had fully recovered at 4 months' follow-up. PMID- 11393119 TI - Video-assisted thoracoscopic surgery for the diagnosis of patients with hilar and mediastinal lymphadenopathy. AB - In areas where tuberculosis (TB) is rare, cases of hilar and mediastinal lymphadenopathy are often attributed to the diagnosis of sarcoidosis or a malignant process. However, these manifestations have been only sparsely reported in countries with high rates of TB. The role of simultaneous lung biopsy in the differential diagnosis of these patients using a thoracoscopic approach is also undetermined. In this prospective study, 15 adult patients with hilar and mediastinal lymphadenopathy were evaluated using video-assisted thoracoscopy during the period from May 1995 through September 1999. Biopsy of the hilar and mediastinal lymph nodes was undertaken in all 15 patients, and a wedge biopsy of the lungs was performed whenever frozen section of the nodes disclosed granulomatous inflammation. The final diagnoses included sarcoidosis (10 patients), TB (2), metastatic small cell carcinoma (2), and reactive lymphoid hyperplasia (1). No morbidity or mortality was associated with the operation. In patients with sarcoidosis and TB, most of the lymph node biopsy specimens disclosed extensive hyaline fibrosis. Lung biopsy specimens presented small non necrotizing granulomas with multinucleated giant cells even in the absence of demonstrable parenchymal lesions. In the two patients with TB, identification of acid-fast bacilli and growth of Mycobacterium tuberculosis occurred only in lung specimens and not in specimens from lymph nodes. Video-assisted thoracoscopic surgery is a safe, simple, and effective procedure for the diagnosis of patients with hilar and mediastinal lymphadenopathy. Our results suggest that for a better differentiation between TB and sarcoidosis, an additional lung biopsy could be undertaken to provide specimens for microscopic examination and culture. PMID- 11393120 TI - Erythrocyte Lewis antigen phenotypes of dyspeptic patients in Taiwan- correlattion of host factor with Helicobacter pylori infection. AB - BACKGROUND AND AIMS: Because of the molecular mimicry between Helicobacter pylori lipopolysaccharide and host Lewis blood-group antigens, Lewis antigen may mediate specific H. pylori binding to surface epithelial cells in gastric mucosa. We, therefore, tested whether different Lewis antigen phenotypes have different prevalence rates of H. pylori infection, and determined the specific H. pylori related disease entities or histologic features. METHOD: A total of 342 dyspeptic patients without previous anti-H. pylori therapy were enrolled after endoscopy. The Lewis phenotypes, defined as Le(a-b-), Le(a-b+), Le(a+b-), and Le(a+b+) subtypes, were determined from the expression or absence of Lewis antigens (Le(a) and Le(b)) on erythrocytes in each patient using monoclonal antibodies. The H. pylori-specific gastric histology was evaluated using the updated Sydney's system. RESULTS: Of 342 patients, 233 (68.1%) had H. pylori infection. The H. pylori infection rates were significantly higher in patients with Lewis phenotypes Le(a+b-) and Le(a+b+) (p < 0.05). Patients expressing the Le(a) antigen had a higher H. pylori infection rate than those without the Le(a) antigen (80.8 vs 64%, p < 0.005). In H. pylori-infected patients, patients expressing Le(b) antigen had a lower rate of gastroduodenal ulcers than those without Le(b) antigen (46.9 vs 61.4%, p < 0.05). H. pylori-positive patients who expressed the Le(b) antigen had higher bacterial density and inflammation severity in the gastric cardia than those who did not. Patients who expressed the Le(a) antigen had lower bacterial density, less chronic inflammation severity, and lower frequency of lymphoid follicles in the gastric cardia than those who did not (p < 0.05). CONCLUSION: The erythrocyte Lewis phenotype can be a significant host factor related to susceptibility, different histologic patterns, and clinical outcomes of H. pylori infection in Taiwan. PMID- 11393121 TI - Comparison of effectiveness of intracellular and extracellular preservation solution on attenuation in ischemic-reperfusion lung injury in rats. AB - BACKGROUND: Human lung allografts can only be preserved for 6 hours. Experimental interventions that reduce ischemia-reperfusion (I/R) lung injury can be used to improve the properties of the preservation solution. The best solution for lung preservation is still a matter of controversy. The purpose of this study was to compare the protective effects of various solutions on I/R lung injury in Sprague Dawley rats. METHODS: The following solutions were compared: a physiological salt solution; an intracellular preservation solution (the University of Wisconsin Solution, UW); an extracellular preservation solution (EP3); and the extracellular preservation solution with the addition of various protective agents--EP3 plus dexamethasone (Dex) (EP3-a), plus glutathione (GLU) and allopurinol (ALL) (EP3-b), and EP3 plus GLU, ALL, lactobionate (LACT), and raffinose (RAF) (EP3-c). I/R lung injury was induced by ischemia for either 45 or 60 minutes, followed by reperfusion for 60 minutes. Hemodynamic changes, lung weight gain (LWG), and capillary filtration coefficients were measured. RESULTS: Both EP3 and UW preservation solutions had partial attenuation effects on I/R lung injury, but UW produced a better attenuation effect than EP3. Use of modified EP3 solutions containing either protective agents (GLU, ALL, or Dex) or impermeants (LACT and RAF) improved the ability of EP3 to reduce I/R lung injury. The LWG using the modified EP3-c solution was the lowest among all groups. UW induced pulmonary hypertension. After I/R challenge, pulmonary arterial pressure with EP3-c was lower than with UW. Based on a lower LWG and the changes in hemodynamics, EP3-c is a better lung preservation solution than UW and EP3. CONCLUSIONS: Based on the attenuation of I/R injury, we conclude that there is no significant difference between intracellular UW and extracellular (EP3-a, EP3-b) preservation solutions in this rat model, but the addition of protective agents and impermeants to the solution are important. The findings suggest that EP3-c might be a better lung preservation solution than UW. PMID- 11393122 TI - Enhanced prostaglandin E2 secretion in sputum from asthmatic patients after zafirlukast therapy. AB - BACKGROUND AND PURPOSE: Leukotrienes are important inflammatory mediators of bronchial asthma that cause bronchoconstriction, mucous secretion, and increased vascular permeability. Current guidelines recommend anti-leukotriene agents as alternative treatments for asthma; however, data on their anti-inflammatory effect is lacking. METHODS: The purpose of this study was to determine the anti inflammatory effect of zafirlukast, a leukotriene antagonist, in patients with bronchial asthma. A total of 30 adult patients with mild persistent asthma received 6 weeks of zafirlukast treatment. Peak expiratory flow rate (PEFR) was determined before and after therapy to assess clinical efficacy. Both serum and sputum samples were collected before and after therapy and concentrations of eosinophil cationic protein (ECP), prostaglandin E2 (PGE2), and leukotriene E4 (LTE4) were measured. RESULTS: A significant improvement in PEFR was found after zafirlukast therapy (p = 0.017). There was also a significant reduction in serum ECP concentration (13.6 +/- 2.4 micrograms/L vs 10.3 +/- 2.1 micrograms/L, p < 0.025) and a significant increase in sputum PGE2 concentration (112.7 +/- 14.0 pg/mL vs 176.8 +/- 32.1 pg/mL, p < 0.01). The percentage eosinophil count and the concentrations of ECP and LTE4 in the sputum were not significantly different after therapy. CONCLUSION: This study found a significant reduction in serum ECP and a significant increase in sputum PGE2 concentrations in asthmatic patients after zafirlukast treatment, both of which were significantly associated with improvement in PEFR. The modulation of PGE2 and ECP production might occur through the anti-inflammatory effect of zafirlukast. PMID- 11393123 TI - Erythema induratum: a clinicopathologic and polymerase chain reaction study. AB - BACKGROUND: In Taiwan, cutaneous lesions with granulomatous lobular panniculitis, with or without vasculitis, are usually diagnosed as erythema induratum (EI), a common form of tuberculid associated with tuberculosis. However, there has been no study to elucidate the extent of this association in Taiwan. The aim of this study was to document the spectrum of the pathologic findings in EI and its association with Mycobacterium tuberculosis. METHODS: The diagnostic/inclusion criteria for EI were recurrent tender subcutaneous nodules on the legs, histopathologic findings of granulomatous lobular or septolobular panniculitis plus necrosis or vasculitis, and positive response to antituberculosis therapy. The clinicopathologic findings in the 12 cases that fulfilled these criteria were analyzed, and nested polymerase chain reaction (PCR) was used to identify M. tuberculosis complex DNA from formalin-fixed, paraffin-embedded sections. RESULTS: Eleven women and one man were included in the study, ranging from 18 to 70 years old (mean, 40.6 yr). The duration of the disease ranged from 10 days to 10 years (mean, 2.1 yr). Histopathology revealed granulomatous panniculitis; a diffuse lobular pattern was observed in nine cases and a focal lobular/septolobular pattern in three. Vasculitis was found in nine cases, four affecting an artery or vein, with three occurring in the patients with focal panniculitis. Ten cases showed various degrees of caseous necrosis. Eosinophils or focal eosinophilia were fairly common (10 patients). From PCR, nine patients were positive for M. tuberculosis complex DNA. CONCLUSIONS: Type III and IV hypersensitivity reactions to M. tuberculosis complex were involved in the pathogenesis of EI. Our results suggest that approximately half of the cases with pathologic findings consistent with EI or nodular vasculitis in Taiwan are associated with M. tuberculosis. PMID- 11393124 TI - Surgical treatment of subluxation and dislocation of the hips in cerebral palsy patients. AB - BACKGROUND AND PURPOSE: Progressive subluxation and dislocation of the hip are major complications in patients with cerebral palsy (CP), causing functional deterioration and difficulties in personal hygiene. Treatment of these problems is difficult and complicated. The purpose of this study was to describe the surgical results and long-term follow-up in a group of CP patients. METHODS: Twenty-three CP patients with subluxated (15 hips) or dislocated hips (12 hips) underwent corrective surgery between 1985 and 1993. This included 11 quadriplegic, eight diplegic, and four hemiplegic patients. Before surgery, four patients were bed-ridden, eight were sitters, six were house-ambulators, and five were community-ambulators. The average age at surgery was 8 years and 5 months. The surgical procedures consisted of femoral varus derotational osteotomy in 21 patients (25 hips), selected soft tissue release in 18 patients (22 hips), and pelvic osteotomy in 18 patients (20 hips). The center-edge angle, acetabular index, and neck-shaft angle were used as parameters to evaluate preoperative and postoperative radiographic changes. RESULTS: After an average follow-up of 4.8 years, 19 patients (22 hips) had gained hip stability, and also had improved functional status. The four bed-ridden patients all became sitters; six of the eight sitters became house-ambulators and one became a community-ambulator; all six house-ambulators became community-ambulators, and the five community ambulators had functional improvement. Complications included nonunion at the femoral osteotomy site in one hip, redislocation in two hips, and resubluxation in one hip. CONCLUSIONS: We conclude that subluxated or dislocated hips in patients with CP can be effectively treated with aggressive correction, which may include soft tissue release, femoral derotational osteotomy, and pelvic osteotomy for improvement of hip range of motion and functional status. PMID- 11393125 TI - First-trimester Down's syndrome screening by fetal nuchal translucency measurement in Taiwan. AB - BACKGROUND: Fetal nuchal translucency (NT) measurement is now widely used in many Western countries as a screening tool for Down's syndrome during the first trimester. However, at present there is no data on its use in Taiwan. The purpose of the present study was to evaluate the efficacy of NT measurement in first trimester Down's syndrome screening in Taiwan. METHODS: We conducted a prospective study from October 1997 to May 1999. Sonographic measurement of fetal NT was performed in 1,249 fetuses at 9-14 weeks of gestation. Transabdominal ultrasound scanning was performed to obtain a sagittal section of the fetus for measuring the crown-rump length (CRL) and the maximum thickness of the subcutaneous translucency between the skin and the soft tissue overlying the cervical spine. Two different cut-off points were used for screening: a fixed cut off point of at least 2.5 mm and a CRL-related cut-off point. In the latter method, fetuses with an NT measurement in the 95th percentile were considered at high risk for Down's syndrome. RESULTS: Three fetuses had Down's syndrome, with NT measurements of 2.1 mm, 2.7 mm, and 4.0 mm. The false positive rates for the fixed cut-off point and CRL-related cut-off point were 6.3% and 4.6%, respectively. Both methods had a sensitivity of 66.7%. However, the screening program using the CRL-related cut-off point had two advantages: a higher specificity (95.5% vs 93.8%) and a more reasonable distribution pattern for screening. CONCLUSION: This study showed that NT measurement is a potential screening tool for Down's syndrome during the first trimester in Taiwan. Using CRL-related cut-off points for screening is more reasonable than using a fixed cut-off point. PMID- 11393126 TI - Comparative study of daily activities of pregnant and non-pregnant women after in vitro fertilization and embryo transfer. AB - BACKGROUND AND PURPOSE: The purposes of this study were to explore daily activity following embryo transfer (ET) in women who had undergone in vitro fertilization (IVF), and to compare the differences between pregnant and non-pregnant cycles. METHODS: This prospective survey was conducted in a medical center in northern Taiwan. Subjects were women who had completed one IVF cycle. Using a structured questionnaire, the subjects were asked whether they had changed their normal daily activities during the 2-week waiting period following ET. Data were collected on the day of ET and on the day of pregnancy testing before the outcome of the treatment was known. After the results of the pregnancy test were known, subjects were divided into pregnant and non-pregnant groups, with 30 subjects in each group. RESULTS: Of the 60 participants, 56 reported that they had tried to rest in bed for more than 2 hours following ET, and 54 reported that they limited their self-care activity on the day of ET. During the 2-week waiting period following ET, 55 walked at a slow pace, 51 limited their social activity, 47 avoided the use of stairs, 45 limited their self-care activity, and 40 adjusted their work load. However, a comparison of daily activity variables between the two groups yielded no significant differences. CONCLUSIONS: Most subjects reported that they restricted their daily activities following IVF/ET, even though their doctors suggested that they return to their normal routines. Bed rest was not correlated with successful IVF. PMID- 11393127 TI - Hereditary nonpolyposis colorectal cancer with gynecologic malignancies: report of two families in Taiwan. AB - Hereditary nonpolyposis colon cancer (HNPCC), also known as Lynch syndrome, is characterized by germline and somatic mutations of DNA mismatch repair genes with dominant inheritance of site-specific colorectal cancer or colorectal cancer plus cancers of extracolonic sites. We describe two Taiwanese HNPCC families with members who had predominantly gynecologic malignancies. In one family, the 53 year-old proband was found to have five synchronous and metachronous tumors of the genitourinary system, which included endometrial adenocarcinoma, cervical squamous cell carcinoma, ureteral and bladder transitional cell carcinoma, and ovarian teratoma. Fourteen of her first- and second-degree relatives were victims of genitourinary and gastrointestinal malignancies. The other family was characterized by four sisters who developed endometrial adenocarcinomas at young ages (36-42 yr). Their father died of both stomach cancer and colon cancer at age 47. The diagnosis of HNPCC was confirmed in this family by genetic analysis. A heterozygous germline mutation (G5 to G6 frame-shift at 183-187) of the hMSH2 (human MutS homolog 2) gene was identified in white blood cells of all the affected family members. The frequent presentation of genitourinary cancers in HNPCC highlights the importance of family-history taking in patients with gynecologic cancers and a genetic diagnosis of HNPCC. PMID- 11393128 TI - Prophylactic total thyroidectomy in an 8-year-old girl with multiple endocrine neoplasia type 2A. AB - We report a genetic carrier of multiple endocrine neoplasia type 2A (MEN 2A) who underwent prophylactic total thyroidectomy. The asymptomatic carrier of MEN 2A, an 8-year-old Taiwanese girl, was admitted for early thyroidectomy. Preoperative basal plasma concentrations of calcitonin and intact parathyroid hormone, and urine vanillylmandelic acid excretion, were normal. Ultrasonography of the thyroid was also normal. Pathology did not reveal any gross lesion but C-cell hyperplasia of the thyroid gland was found microscopically. Prophylactic total thyroidectomy is encouraged and is justified during the first decade of life for MEN 2A carriers. PMID- 11393129 TI - Multiple myeloma associated with extramedullary plasmacytoma causing nerve root compression: a case report. AB - Multiple myeloma associated with extramedullary plasmacytoma at initial presentation is rare. We describe a 45-year-old female patient with an initial presentation of low back pain and right side L5, S1 radiculopathy. There was no evidence of vertebral involvement but an epidural tumor was found later during neurosurgical intervention. The final diagnosis was immunoglobulin G, kappa multiple myeloma complicated with spinal root compression by an extramedullary plasmacytoma. No osteolytic lesion was noted over the length of the spine. Pathology revealed high-grade plasmablastic myeloma. During the clinical course, the patient was refractory to induction chemotherapy, and there was progressive deterioration of renal function. Urinary tract infection by Morganella morganii and pulmonary infection of unknown cause developed 5 months later, and the patient died. PMID- 11393130 TI - Chronic granulomatous disease: a case report. AB - Chronic granulomatous disease is one form of the phagocyte function disorder. Unlike most patients with chronic granulomatous disease who develop signs and symptoms of chronic and recurrent pyogenic infections during the first 2 years of life, patients with mild forms of the disease may not present until the teenage years or even adulthood. Thus, the diagnosis in these mild-form patients is often delayed. This paper describes a patient with the mild form of chronic granulomatous disease. A 7-year-old boy was admitted to our ward with intermittent high fever and a left neck mass present for about 1 week. He had a history of persistent infection in the bilateral lower face lasting for about 1 year during his fourth year of life. Family history was unremarkable except that the patient's elder sister had a history of persistent oral mucosal wound infection for about 1 year during the fifth year of life. On physical examination, there were scars over the patient's bilateral lower face. Bacterial culture of pus drained from the neck mass revealed Burkholderia cepacia, a rare species in patients without immunodeficiency. A series of antibiotics, including oxacillin, clindamycin, and piperacillin, was given, and two incision operations for drainage and debridement were performed. The neck mass resolved completely about 1.5 months later. This history indicated that the patient might have chronic granulomatous disease. A definite absence of superoxide activity in the patient's granulocytes detected by chemiluminescence and nitroblue tetrazolium dye test confirmed this diagnosis. PMID- 11393131 TI - Pseudocyst formation after intertrochanteric fracture fixation: a case report. AB - We describe a 62-year-old woman who developed two pseudocysts, 25 x 15 cm and 20 x 12 cm, in the left proximal thigh as a complication 19 years after internal fixation of an intertrochanteric fracture. She received a 135 degrees dynamic hip screw and side plate in May 1979. She continued to live at home without major discomfort until May 1997. Two huge pseudocysts were noted in the left proximal thigh without trauma history. Angiography was normal. Computerized tomography scan revealed two voluminous cystic lesions without septa in the left proximal thigh, with accumulated fluid. During surgery, two huge cysts were found in the left proximal thigh, and their orifices were found slightly proximal to the curvature of the side plate. The pathology showed that the cysts consisted of a nonepithelialized wall of granulation tissue compatible with a pseudocyst. The patient had no further problems 2 years after surgery. We found no reports in the literature of this rare complication. The development of the pseudocysts may have been the result of chronic low-grade trauma due to irritation between the soft tissue and the implant. Orthopedic surgeons should be aware of the possible development of this rare complication following internal fixation of an intertrochanteric fracture. PMID- 11393132 TI - Characteristics of coronary artery blood flow waveforms and the mechanism of myocardial ischemia in aortic valve disease. PMID- 11393133 TI - Left ventricular function and autonomic nervous system balance during two different stages of the menstrual cycle. PMID- 11393134 TI - Eccentric mitral regurgitation can imitate a diagnosis of pneumonia. PMID- 11393135 TI - Cardiac dysautonomia in patients with superior vena cava syndrome due to compression by lung cancer. PMID- 11393136 TI - Double right coronary artery. PMID- 11393137 TI - A concealed complication of primary angioplasty: coronary fistula. PMID- 11393138 TI - Ventricular repolarization changes during thyrotropin releasing hormone test. PMID- 11393139 TI - Measurement of potassium concentration in salivary and sweat fluids as a screening test for the long QT1 syndrome. PMID- 11393140 TI - Tachycardia cardiomyopathy. PMID- 11393141 TI - Photo quiz. Round pneumonia due to Streptococcus pneumoniae. PMID- 11393142 TI - Bibliography. Current world literature. Brain and nervous system. PMID- 11393143 TI - Bibliography. Current world literature. Head and neck. PMID- 11393144 TI - Bibliography. Current world literature. Genitourinary system. PMID- 11393145 TI - A normative reference database study for Pronosco X-posure System. AB - Cortical width from radiographs has been used for more than 40 yr as a means of estimating bone strength. In the last 5-10 yr, increased availability of computers and the development of automated algorithms for image assessment have led to an increased interest in radiogrammetry. In this study, we examined a new radiogrammetry device, the Pronosco X-posure System, which estimates bone mineral density (BMD) from forearm/hand radiographs. We obtained hand and forearm radiographs and performed dual X-ray absorptiometry (DXA) at the wrist and hip on 832 women ages 20-79 at four clinical centers across the United States. We then used the X-posure System to estimate BMD (DXR-BMD). The goal of the study was to establish reference ranges for the method and to compare the measurement to DXA measurements of BMD at the wrist and hip. Using statistical models, we estimated that the peak value for DXR-BMD occurred at age 38 (mean = 0.598 g/cm2, standard deviation = 0.034 g/cm2). The correlation between DXR-BMD and DXA was 0.90 at the wrist and 0.61 at the hip. The relationship of DXR-BMD to reported history of fracture was of similar magnitude to that for DXA at the wrist and hip. The strong correlation of DXR-BMD from the X-posure System with DXA at the wrist from the Hologic machine suggests that the X-posure System may be an alternative to DXA at the wrist for the assessment of osteoporosis. PMID- 11393146 TI - Body composition of professional football (soccer) players determined by dual X ray absorptiometry. AB - A three-compartment body composition analysis of 42 professional football (soccer) players and 33 age- and body mass index-matched control subjects was determined by dual X-ray absorptiometry (DXA). The equipment provided a direct measurement of fat, lean, and bone mass. Fat mass was significantly higher in the controls subjects whereas lean mass and bone mass were markedly higher in the players. The percentage of body weight fat varied from 6.1 to 19.5% in the football players and from 9.1 to 29.9% in the control subjects. The respective averages were 12.0 +/- 3.1 and 19.2 +/- 5.6% (p < 0.001). The midfielders had a significantly higher percentage of fat (13.6 +/- 3.3%) than backs or forwards (11.1 +/- 2.8 and 11.0 +/- 2.3%, p < 0.05 and p < 0.06, respectively). In the football players, the correlation between age and fat mass was significant (r = 0.53, p < 0.001), whereas there was no correlation between fat and age in the control subjects (r = 0.13 p > 0.1). This article provides, for the first time, DXA analysis of body composition of football players in relation to their age and function. The results should be of interest to coaches because they will help improve athletes' performance. PMID- 11393147 TI - Iatrogenic keratectasia after laser in situ keratomileusis. PMID- 11393148 TI - Convertible mini-nuc technique. PMID- 11393150 TI - Diffuse lamellar keratitis: are meibomian secretions responsible? PMID- 11393149 TI - Removal of an ophthalmic viscoelastic device. PMID- 11393151 TI - History of Harvard/MGH Oral and Maxillofacial Surgery. PMID- 11393152 TI - TDA directory and membership roster. 2000-2001. PMID- 11393153 TI - Dealing the CARDs between life and death. AB - Exciting evidence presented at a recent meeting shows that proteins containing CARD, or related DD or DED, motifs are centrally involved in assembling protein complexes that drive activation of either IkappaB kinase or caspases by facilitating intermolecular juxtapositioning. Thus, CARD-family proteins occupy crucial positions in divergent stress-associated signalling pathways that culminate in inflammatory responses or apoptosis. PMID- 11393154 TI - Disease nomenclature. PMID- 11393155 TI - Veterinary parasitology in the 21st Century. PMID- 11393156 TI - Control of trypanosomes in cattle,. PMID- 11393157 TI - Two different types of apoptosis in thymocytes. AB - Thymocyte cell death was investigated after UVA-irradiation (365 nm) of the primary thymocyte culture in vitro. To determine the mode of cell death two fluorescent DNA-binding dyes (Acridine Orange and Ethidium Bromide) were used along with flow cytometry. Thymocytes undergo apoptosis spontaneously, but a percentage of apoptotic cells was seen to increase in a dose dependent manner after thymocytes exposition to 1-100 J/m2 UVA. These doses are lower than those commonly used for apoptosis induction in other lymphoid cells. Using flow cytometry, we demonstrated that UVA-irradiation induced two different types of apoptosis. The one referred to as the "fast" apoptosis was recorded within 1-4 h following irradiation, whereas the other one, called the "delayed" apoptosis occurred within 3-24 h after irradiation. After UVA-irradiation, the activation of the former prevented the development of the latter; whereas the inhibition of delayed apoptosis brought about the induction of the fast apoptosis in thymocytes. The interrelation between the fast and delayed types of apoptosis in thymocytes can be modulated by cycloheximide, an inhibitor of protein synthesis. PMID- 11393158 TI - Checking compatibility of pulmonary artery catherers and percutaneous sheath introducers. PMID- 11393159 TI - Emergence delirium: statistically significant or not? PMID- 11393160 TI - National Nutrition Summit, May 30, 2000: results of the obesity discussion groups. PMID- 11393161 TI - Backseat driving? Accessing phosphate beyond the rhizosphere-depletion zone. PMID- 11393162 TI - Do plants have more genes than humans? PMID- 11393163 TI - Chronomics complement, among many other fields, genomics and proteomics. PMID- 11393164 TI - An overview of molecular diagnosis of steroid 21-hydroxylase deficiency. PMID- 11393165 TI - The immune response to ocular herpes simplex virus type 1 infection. AB - Herpes simplex virus type 1 (HSV-1) is a prevalent microbial pathogen infecting 60% to 90% of the adult world population. The co-evolution of the virus with humans is due, in part, to adaptations that the virus has evolved to aid it in escaping immune surveillance, including the establishment of a latent infection in its human host. A latent infection allows the virus to remain in the host without inducing tissue pathology or eliciting an immune response. During the acute infection or reactivation of latent virus, the immune response is significant, which can ultimately result in corneal blindness or fatal sporadic encephalitis. In fact, HSV-1 is one of the leading causes of infectious corneal blindness in the world as a result of chronic episodes of viral reactivation leading to stromal keratitis and scarring. Significant inroads have been made in identifying key immune mediators that control ocular HSV-1 infection and potentially viral reactivation. Likewise, viral mechanisms associated with immune evasion have also been identified and will be discussed. Lastly, novel therapeutic strategies that are currently under development show promise and will be included in this review. Most investigators have taken full advantage of the murine host as a viable working in vivo model of HSV-1 due to the sensitivity and susceptibility to viral infection, ease of manipulation, and a multitude of developed probes to study changes at the cellular and molecular levels. Therefore, comments in this review will primarily be restricted to those observations pertaining to the mouse model and the assumption (however great) that similar events occur in the human condition. PMID- 11393166 TI - New concepts in characterization of ischemically injured myocardium by MRI. AB - New concepts regarding the assessment of ischemic myocardial injuries have been addressed in this Minireview using magnetic resonance imaging (MRI). MRI, with its different techniques, brings not only anatomic, but also physiologic, information on ischemic heart disease. It has the ability to measure identical parameters in preclinical and clinical studies. MRI techniques provide the ideal package for repeated and noninvasive assessment of myocardial anatomy, viability, perfusion, and function. MR contrast agents can be applied in a variety of ways to improve MRI sensitivity for detecting and assessing ischemically injured myocardium. With MR contrast agents protocol, it becomes possible to identify ischemic, acutely infarcted, and peri-infarcted myocardium in occlusive and reperfused infarctions. Necrosis specific and nonspecific extracellular contrast enhanced MRI has been used to assess myocardial viability. Contrast-enhanced perfusion MRI can explore the disturbances in large (angiography) and small coronary arteries (myocardial perfusion) as the underlying cause of myocardial dysfunction. Perfusion MRI has been used to measure myocardial perfusion (ml/min/g) and to demonstrate the difference in transmural myocardial blood flow. Information on no-reflow phenomenon is derived from dynamic changes in regional signal intensity after bolus injection of MR contrast agents. Another development is the near future availability of blood pool MR contrast agents. These agents are able to assess microvascular permeability and integrity and are advantageous in MR angiography (MRA) due to their persistence in the blood. Noncontrast enhanced MRI such as cine MRI at rest/stress, sodium MRI, and MR spectroscopy also have the potential to noninvasively assess myocardial viability in patients. Futuristic applications for MRI in the heart will focus on identifying coronary artery disease at an early stage and the beneficial effects of new therapeutic agents such as intra-arterial gene therapy. MR techniques will have great future in the drug discovery process and in testing the effects of drugs on myocardial biochemistry, physiology, and morphology. Molecular imaging is going to bloom in this decade. PMID- 11393167 TI - Alpha-fetoprotein structure and function: relevance to isoforms, epitopes, and conformational variants. AB - Mammalian alpha-fetoprotein (AFP) is classified as a member of the albuminoid gene superfamily consisting of albumin, AFP, vitamin D (Gc) protein, and alpha albumin. Molecular variants of AFP have long been reported in the biomedical literature. Early studies identified isoelectric pH isoforms and lectin-binding variants of AFP, which differed in their physicochemical properties, but not in amino acid composition. Genetic variants of AFP, differing in mRNA kilobase length, were later extensively described in rodent models during fetal/perinatal stages, carcinogenesis, and organ regeneration. With the advent of monoclonal antibodies in the early 1980s, multiple antigenic epitopes on native AFP were detected and categorized, culminating in the identification of six to seven major epitopes. During this period, various AFP-binding proteins and receptors were reported to inhibit certain AFP immunoreactions. Concomittantly, human and rodent AFP were cloned and the amino acid sequences of the translated proteins were divulged. Once the amino acid composition of the AFP molecule was known, enzymatic fragments could be identified and synthetic peptide segments synthesized. Following discovery of the molten globule form in 1981, the existence of transitory, intermediate forms of AFP were acknowledged and their physiological significance was realized. In the present review, the various isoforms and variants of AFP are discussed in light of their potential biological relevance. PMID- 11393168 TI - Genetics of arterial prothrombotic risk states. AB - Coronary artery disease is a leading cause of death worldwide and the largest killer of men and women in the United States. The pathophysiology of myocardial infarction is multifactorial, and numerous physiologic systems converge to dictate the formation of the two fundamental lesions, thrombosis and atherosclerosis. In this review we address genetic aspects of arterial thrombosis and the key thrombotic factors that have been associated with the increased risk for its development. Specifically, we consider components of coagulation, fibrinolysis, and platelet adhesive receptors, and we review the genetic epidemiology and in vitro laboratory data regarding their risk for the acute coronary syndromes. In combination with traditional risk factor assessment, in the near future these inherited markers can be used to manage patients with vascular disease through a better utilization of invasive or expensive diagnostic testing, as well as pharmacologic intervention. PMID- 11393169 TI - Cholangiohepatitis and inflammatory bowel disease induced by a novel urease negative Helicobacter species in A/J and Tac:ICR:HascidfRF mice. AB - Helicobacter bilis and H. hepaticus, both urease-positive intestinal helicobacters of mice, have been shown experimentally to induce proliferative typhlocolitis in scid mice. We recently isolated a urease-negative Helicobacter sp. (H. sp.) that also induced proliferative typhlocolitis in pilot studies in scid mice. To determine the pathogenic potential of H. sp. in immunocompromised and immunocompetent mice, 5-week old male A/J or Tac:Icr:Ha(ICR)-scidfRF mice were inoculated by intraperitoneal (IP) injection with approximately 3 x 10(7) colony-forming units (CFU) of H. sp. Mice were necropsied at various time points postinoculation (PI). Sham-inoculated mice had no clinical, gross, or histopathological lesions. In contrast, scid mice inoculated IP with H. sp. had severe hemorrhagic diarrhea and decreased weight gain at 2, 7, and 18 weeks postinoculation (PI), with severe proliferative typhlocolitis, phlebothrombosis, and hepatitis. A/J mice had no clinical signs, but had mild to moderate proliferative typhlocolitis and moderate to marked cholangiohepatitis at 7 and 24 weeks PI. A/J mice infected with H. sp. developed robust immune responses of a predominant Th1 type. This report demonstrates that infection with a urease negative helicobacter can cause inflammatory bowel disease (IBD) and hepatitis in scid and immunocompetent A/J mice. These results provide a new model of IBD and cholangio-hepatitis associated with a specific urease-negative, novel H. species. PMID- 11393170 TI - Suppression of the development of experimentally induced uterine adenomyosis by a novel matrix metalloproteinase inhibitor, ONO-4817, in mice. AB - The inhibitory effects of a novel, orally active matrix metalloproteinase (MMP) inhibitor, ONO-4817, on the development of uterine adenomyosis induced experimentally by pituitary grafting were examined in mice. Mice were given transplants of isologous anterior pituitary glands (PGs) into the right uterine lumen at 7 weeks of age and were fed chow containing 0.1% to 1.0% ONO-4817 from 8 to 14 weeks of age. Mice treated with 0.3% or 1.0% ONO-4817 showed a significantly lower incidence of the development of adenomyosis than vehicle treated mice. To evaluate the inhibitory effects of ONO-4817 on the progression of the invasion of the adenomyotic tissues, mice receiving PG grafts at 7 weeks of age were treated with 1.0% ONO-4817 from 13 to 17 weeks of age. The degree of pathological progression of adenomyosis was graded from 1 to 5 in increments of 1. The degree of the progression of the lesion was less in the uteri exposed to ONO-4817 (2.71 +/- 0.93) than in the uteri not exposed to the inhibitor (4.33 +/- 0.75). Finally, the invasiveness of endometrial stromal cells obtained from adenomyotic uteri into Matrigel consisting mainly of type IV collagen and laminin was examined using an invasion assay. The assay showed that the treatment with ONO-4817 markedly suppressed the invasion of the stromal cells of the adenomyotic uteri into the gel. These results indicate that ONO-4817 may be an effective inhibitor of the development of adenomyosis. PMID- 11393171 TI - Impaired release of ATP from red blood cells of humans with primary pulmonary hypertension. AB - Previously, we reported that in the isolated perfused rabbit lung, red blood cells (RBCs) obtained from either rabbits or healthy humans were a required component of the perfusate to unmask evidence of nitric oxide (NO) participation in regulation of the pulmonary circulation. In addition, we found that mechanical deformation of rabbit and healthy human RBCs released ATP, a known agonist for enhanced NO synthesis. In contrast, RBCs obtained from patients with cystic fibrosis (CF) did not release ATP in response to mechanical deformation. The coexistence of airway disease and alveolar hypoxia in patients with CF precluded the drawing of conclusions relating a defect in RBC ATP release with the pulmonary hypertension associated with CF. Airway disease and alveolar hypoxia are not, however, features of primary pulmonary hypertension (PPH), a human condition of unknown etiology. We postulated that a defect in NO generation might contribute to the increased pulmonary vascular resistance in PPH, and as a first step, we hypothesized that RBCs obtained from patients with PPH would not release ATP. In contrast to RBCs of healthy humans, when RBCs of PPH patients were passed through filters (average pore size 12, 8, or 5 microm), ATP was not released and the RBCs exhibited reduced deformability. Moreover, when incubated with the active cAMP analogue, Sp-cAMP (100 microM), an activator of the CF transmembrane conductance regulator, ATP was not released. These results demonstrate that RBCs obtained from patients with PPH fail to release ATP whether the stimulus is mechanical or pharmacological. Thus, failure of RBCs to release ATP in patients with PPH might be a major pathogenetic factor that accounts for the heretofore unknown etiology of their pulmonary hypertension. PMID- 11393172 TI - VLCD-induced weight loss improves heart rate variability in moderately obese Japanese. AB - To evaluate the effects of weight reduction on the autonomic nervous system in obese patients, we investigated heart rate variability (HRV) based on 24-hr ambulatory electrocardiogram (ECG) recordings before and after weight reduction. To aim for weight reduction, 16 obese patients were treated with the very-low calorie conventional Japanese diet (VLCD-CJ) therapy combined with behavior therapy. Percent weight reduction was 17.8% +/- 1.5% (means +/- SEM), but mean blood pressure did not change significantly after VLCD-CJ therapy. The mean normal R-R interval (mNN) of the 24-hr ECG and all other five time-domain indices increased after weight reduction. Spectral analysis revealed that weight reduction increased the high frequency (HF) component, but decreased the ratio of low to high (LF/HF) components. Rate of change in mNN or HF correlated positively with reduction rate of body mass index, but not that in LF/HF. Analysis of daily fluctuations in each HRV parameter showed that significant improvement after weight loss occurred mainly during the nocturnal period, but an HF component was improved throughout the day and night periods. These findings indicate that functional impairment of the autonomic nervous system in obese subjects, particularly in the nocturnal period, is improved by effective weight reduction after VLCD-CJ therapy. PMID- 11393173 TI - Plasma drug levels compared with DNA incorporation of 3'-azido-3'-deoxythymidine (AZT) in adult cynomolgus (Macaca fascicularis) monkeys. AB - Zidovudine (3'-azido-3'-deoxythymidine, AZT), widely used for the therapy of the Human Immunodeficiency Virus-1 (HIV-1), is a nucleoside analog of thymidine that becomes phosphorylated and incorporated into nuclear and mitochondrial DNA. Levels of AZT incorporation into DNA of humans, monkeys, and mice are highly variable and suggest interindividual variability in phosphorylation pathways. In addition, studies in rhesus monkeys (1) have shown a lack of correlation between levels of unbound AZT in plasma and tissue AZT-DNA. However, the correlation between plasma AZT and tissue AZT-DNA has not been previously examined in the same primate. Here we examine the relationship between AZT-DNA incorporation in leukocytes and multiple organs, and levels of the drug circulating in plasma of adult female cynomolgus (Macaca fascicularis) monkeys. Three monkeys were dosed with 40.0 mg of AZT/day for 30 days by naso-gastric intubation. The average daily dose of 9.9 mg of AZT/kg/body wt was similar to the approximately 8.6 mg of AZT/kg/body wt (600 mg/day) given to adult HIV-1-infected patients. In all three monkeys, at the time of sampling, values for AZT concentrations in plasma were similar and values for AZT incorporation into leukocyte DNA (86.1, 100.0, and 114.1 molecules of AZT/10(6) nucleotides) were also similar. AZT-DNA incorporation was detected in liver, uterus, spleen, and kidney from the three AZT-exposed animals, with values for positive samples ranging from 5.8 to 97.4 molecules of AZT/10(6) nucleotides. In brain cortex and lung DNA from AZT-exposed animals, AZT incorporation was undetectable. The data suggest that organ-specific differences in AZT uptake and/or metabolism may contribute to AZT phosphorylation and subsequent drug incorporation into DNA. In addition, AZT-DNA levels in monkey organs were similar to or lower than values observed in peripheral leukocytes of adult AIDS patients. PMID- 11393174 TI - High glucose concentrations induce oxidative damage to mitochondrial DNA in explanted vascular smooth muscle cells. AB - Oxidative stress is considered to be one of the mechanisms leading to atherosclerosis. It occurs in response to injury or to altered metabolic state. Alterations in cell growth (proliferation or apoptosis) can also contribute to the pathogenesis of atherosclerosis and is influenced by oxidative stress. Smooth muscle cells (SMC) from aortic explants of JCR:LA-cp homozygous cp/cp corpulent rats who are genetically predisposed to develop atherosclerosis exhibit increased SMC proliferation, which can be attenuated by exercise and food restriction. This study was conducted to characterize the effects fo oxidative stress and high glucose media on cell growth and its relationship to mitochondrial DNA integrity and gene expression in explanted aortic SMC from corpulent and lean JCR:LA-cp rats. The results show that SMC from the cp/cp rat appear to be resistant to oxidant-induced cell death and that they accumulate mitochondrial DNA mutations, probably as a result of a reduction in apoptosis. These data suggest that susceptibility to age- and glucose-related atherosclerosis may be related to alterations in redox signaling. PMID- 11393175 TI - Genomic growth hormone gene polymorphisms in native Chinese chickens. AB - Chicken growth hormone (cGH), a polypeptide hormone synthesized in and secreted by the pituitary gland, is involved in a wide variety of physiological functions such as growth, body composition, egg production, aging, and reproduction. Chicken growth hormone polymorphisms have been reported to be associated with certain phenotypes. Our objective is to investigate the GH gene polymorphism in selected strains of native Chinese chickens. Yellow Wai Chow GH gene was characterized by sequencing and was found to have one silent substitution, 31 insertions, and other substitutions spread among the introns. In addition, a novel Mspl site has been identified and characterized in the first intron. Allele frequencies of the intron 1 polymorphism were characterized among 28 populations of native Chinese chickens. Thus, polymorphism of the cGH gene may be useful in phylogenetic analysis, as well as in the design of breeding programs. PMID- 11393176 TI - Immunocytochemical localization of aquaporin-1 in bovine corneal endothelial cells and keratocytes. AB - For immunocytochemistry, cultured bovine corneal endothelial cells (CBCEC) and bovine corneal cryosections were utilized. Preparations were fixed, permeabilized, and incubated with primary rabbit anti-rat aquaporin 1 (AQP1) antibody followed by rhodamine-conjugated secondary antibody, and were counter stained with Sytox nuclear acid stain. Confocal microscopy of CBCEC in the x, y, and z planes showed rhodamine fluorescence, indicating the presence of AQP1 antibody localized to the apical and basolateral domains of the plasma membrane, but not to the membranes of intracellular compartments or other subcellular locations. Preabsorption with control antigenic peptide yielded no positive staining. Similar results were obtained using freshly dissected bovine corneas; in addition, these images showed AQP1 distributed to the plasma membranes of keratocytes. No AQP1 staining was seen in corneal epithelium, and no staining was observed in CBCEC layers exposed to AQP3, AQP4, and AQP5 antibodies. PMID- 11393177 TI - Orexin-A regulates body temperature in coordination with arousal status. AB - Orexins, hypothalamic neuropeptides, are involved in modulation of food intake and arousal status. To further examine their physiological roles in brain function, the effect of centrally administered orexin-A on body temperature was investigated in rats. Assessed by a telemetry sensor system implanted into the abdominal cavity, infusion of orexin-A into the third cerebroventricle (i3vt) increased body temperature in a dose-responsive manner. Expression of uncoupling protein 1 (UCP1) mRNA in brown adipose tissue (BAT), as a marker for peripheral thermogenesis, failed to increase after the infusion. Expression of UCP3 mRNA in skeletal muscle was up-regulated, whereas UCP2 in white adipose tissue was unchanged after the infusion. The resulting information indicates that orexin neurons regulate body temperature in coordination with arousal status independently of peripheral thermogenesis, which is regulated by BAT UCP1. PMID- 11393178 TI - Effects of the xenoestrogen bisphenol A on expression of vascular endothelial growth factor (VEGF) in the rat. AB - Bisphenol-A (BPA) is used to produce polymers for production of polycarbonate and epoxy resins that are used in food containers and dental appliances. BPA binds to estrogen receptors and induces estrogenic activity in a number of biological systems. We recently reported that although Fisher 344 (F344) and Sprague-Dawley (S-D) rat strains exhibit different sensitivities to BPA at the level of vaginal epithelial cell proliferation, there was no difference in immediate early proto oncogene expression between the two animal strains. In the present study we investigated the effects of BPA on expression of another estrogen-target gene, vascular endothelial growth factor (VEGF), in the uterus, vagina, and pituitary of F344 and S-D rats. Adult rats were ovariectomized and treated with BPA by intraperitoneal injection at concentrations of 0.02 to 150 mg/kg body wt. Expression of VEGF was monitored by RNase protection assay at 2 hr after treatment. There was a significant effect of dose of BPA on the type of VEGF isoform expressed in the uterus, vagina, and pituitary. BPA induced greater (P < 0.01) levels of VEGF164 and VEGF120+188 than VEGF110 levels. The lowest BPA dose that had a significant (P< 0.05) effect on VEGF expression compared with vehicle treatment was 37.5 mg/kg body wt.; dose-response curves did not differ between strains. This is the first report that the primary response of the uterus, vagina, and pituitary to BPA includes rapid induction of VEGF expression. Due to the capacity of VEGF to engage pleiotropic signaling pathways in other cellular systems, we suggest that modulation of VEFG may play a role in establishing the response of estrogen-target organs to estrogenic xenobiotics. PMID- 11393179 TI - Pathogen population dynamics: the age of the strain. PMID- 11393180 TI - Multicellular resistance: biofilms. PMID- 11393181 TI - Chemotaxis genes, fibrils and PE taxis in Myxococcus xanthus. PMID- 11393182 TI - [Activity of aminopeptidase and inter-alpha-trypsin inhibitor in serum of women in physiological labor]. AB - Aminopeptidase activity was measured towards L-alanine and L-leucine naphtylamides in the blood serum obtained from the non-pregnant women and from women during the stages of the physiological delivery. In the same samples the concentration of inter-alpha-trypsin inhibitor (alpha ITI) was measured. The highest aminopeptidase activity was observed in the stage II, and it was statistically significant comparing to the stage I, whereas the statistically significant increase in the alpha ITI concentration was noticed in stage I in primiparae and in stage I in multiparae. PMID- 11393183 TI - [Bisphosphonates in osteoporosis. Significantly reduces risk of fracture]. PMID- 11393184 TI - [Marketing potential of the medical practice]. AB - Since the 1970s, market forces and cost containment have stimulated a range of economic changes for health care providers. There is increasing pressure for medical practices and other health care organizations to provide simultaneously excellent clinical, financial and satisfaction outcomes. To fulfill these expectations, two central aspects have been identified: customer focus and involvement of all members of a medical organization. Therefore, it is essential to understand the patient's perspective and, where necessary, increase quality of care from the customer's viewpoint. Generally, quality as defined by the customer and by the medical professionals differ substantially. In this article, we present a check-list to evaluate the essential aspects of a medical practice from a patient's view which, in turn, will help to determine which actions have to be taken. Some of the aspects covered by the checklist include satisfaction with care delivered, service performed, warmth of the employees, office's facilities, and physician's behavior. PMID- 11393185 TI - Impact of stereotactic large-core needle biopsy on diagnosis and surgical treatment of nonpalpable breast cancer. AB - INTRODUCTION: Stereotactic large-core needle biopsy is increasingly replacing needle-localized breast biopsy for the diagnosis of nonpalpable breast disease. In this prospective study, the impact of the introduction of this technique on diagnosis and surgical treatment of nonpalpable breast cancer was assessed in two hospitals in The Netherlands. PATIENTS AND METHODS: A total of 84 patients with nonpalpable breast cancer, diagnosed by means of stereotactic large-core needle biopsy (needle biopsy group) were compared with 80 patients diagnosed with nonpalpable breast cancer before the introduction of large-core needle biopsy. These patients were diagnosed by means of needle-localized open breast biopsy (control group). Clinical outcome measures evaluated included: duration of diagnostic and therapeutic intervals and number of surgical procedures required for complete surgical treatment. Subgroup analysis was performed for the category of microcalcifications without tissue distortion. RESULTS: For the needle biopsy group, the median interval between initial referral to the surgeon and the availability of histological diagnosis was 9 days and the interval between initial referral and complete surgical treatment was 31 days. These intervals were significantly longer for the control group (19 days and 44 days respectively); 75% of patients in the needle biopsy group were treated in a single step surgical procedure compared to 16% of the patients in the control group (67 vs 25% respectively for the subgroup). The mean number of surgical procedures required to complete surgical treatment was 1.31 for needle biopsy group vs 1.91 for the open biopsy group (1.46 vs 1.84 for the subgroup). CONCLUSION: Introduction of stereotactic large-core needle biopsy leads to a reduction of the time to diagnosis and the time to complete surgical treatment of nonpalpable breast cancer. It also reduces the number of surgical procedures required for complete surgical treatment of nonpalpable breast cancer. The benefits of large-core needle biopsy may also be anticipated for patients with microcalcifications without tissue distortion. PMID- 11393186 TI - How I do it--Replacement of the oesophagus with colon interposition graft based on the inferior mesenteric vascular system. PMID- 11393188 TI - Sourcebook 2000-2001. PMID- 11393187 TI - Parathyroid carcinoma: diagnosis and management. AB - Parathyroid carcinoma is a rare and difficult diagnosis to make based on the histological features alone. We review five cases of parathyroid carcinoma in the past 30 years and the clinical and biochemical features that facilitate the making of the diagnosis. A favourable outcome can be expected with adequate surgical treatment. PMID- 11393189 TI - Practical ethics. How much to tell a recruit. PMID- 11393190 TI - Pathologic and physiologic interactions of bacteria with the gastrointestinal epithelium. AB - Communication between microorganisms and the gastrointestinal epithelium, ie, bacterial-epithelial "crosstalk," is examined. Because most basic research on the molecular interaction of bacteria with the gut epithelium relates to pathogen enterocyte interaction, crosstalk with pathologic bacterial is considered in detail. Through their interactions with the intestinal epithelium, pathogens can modify epithelium function to enhance their penetration across the epithelial barrier and to exploit mucosal host defenses for their own benefit. Three representative pathogens are used to illustrate the various adaptive techniques used to colonize and penetrate the mucosal barrier. Salmonella enterica typhimurium interacts with the physiologic receptor for epidermal growth factor to co-opt the receptor's signal transduction mechanisms. Enteropathic Escherichia coli secretes a receptor (type III secretion) into the microvillus surface of enterocytes that disrupts the microvillus and alters its actin structure to form a dome-like anchoring site. Shigella flexneri is used to illustrate how pathogens use the follicular epithelial cell (M cell), the physiologic conduit for antigens to reach gut associated-lymphoid tissues, for penetration of the epithelial barrier. Shigella organisms attached to M cells use their endocytotic properties to enter the cell. Once inside the cell, the organism lyses the endocytic vacuole and co-opts actin and myosin to form a propelling tail for further penetration of the epithelium through the basolateral surface. Probiotics can protect the intestine by competing with pathogens for attachment, strengthening tight junctions between enterocytes, and enhancing the mucosal immune response to pathogens. However, additional molecular studies are needed to define more precisely the mechanism of probiotic-epithelial crosstalk. PMID- 11393191 TI - Microbial modulation of innate defense: goblet cells and the intestinal mucus layer. AB - The gastrointestinal epithelium is covered by a protective mucus gel composed predominantly of mucin glycoproteins that are synthesized and secreted by goblet cells. Changes in goblet cell functions and in the chemical composition of intestinal mucus are detected in response to a broad range of luminal insults, including alterations of the normal microbiota. However, the regulatory networks that mediate goblet cell responses to intestinal insults are poorly defined. The present review summarizes the results of developmental, gnotobiotic, and in vitro studies that showed alterations in mucin gene expression, mucus composition, or mucus secretion in response to intestinal microbes or host-derived inflammatory mediators. The dynamic nature of the mucus layer is shown. Available data indicate that intestinal microbes may affect goblet cell dynamics and the mucus layer directly via the local release of bioactive factors or indirectly via activation of host immune cells. A precise definition of the regulatory networks that interface with goblet cells may have broad biomedical applications because mucus alterations appear to characterize most diseases of mucosal tissues. PMID- 11393192 TI - Probiotics in human disease. AB - Western civilization is facing a progressive increase in immune-mediated, gut related health problems, such as allergies and autoimmune and inflammatory diseases, and genetic factors are an unlikely explanation for these rapid increases in disease incidence. Two environmental factors that relate to the modern lifestyle in Western societies are hygiene and nutrition. There has been a decline in the incidence of microbial stimulation by infectious diseases as a result of improved hygiene, vaccination, and antimicrobial medication. In the past, methods of food preservation involved either the natural fermentation or drying of foods; thus, the human diet once contained several thousand times more bacteria than it does today. The development of probiotic, functional foods aims to "kill two birds with one stone," which is accomplished by providing a microbial stimulus to the host immune system by means of beneficial live microorganism cultures that are characteristic of the healthy, human gut microflora, ie, probiotics. Probiotic bacteria were shown to reinforce the different lines of gut defense, which are immune exclusion, immune elimination, and immune regulation. They were also shown to stimulate nonspecific host resistance to microbial pathogens, thereby aiding in pathogen eradication. Consequently, the best documented clinical application of probiotics is in the treatment of acute diarrhea. In humans, documented effects were reported for the alleviation of intestinal inflammation, normalization of gut mucosal dysfunction, and down-regulation of hypersensitivity reactions. These data show that probiotics promote endogenous host defense mechanisms. Thus, modification of gut microflora by probiotic therapy may offer a therapeutic potential in clinical conditions associated with gut-barrier dysfunction and inflammatory response. PMID- 11393193 TI - Clinical applications of probiotic agents. AB - In the past century the beneficial roles of nonpathogenic bacteria in the intestinal lumen were described. In the past decade there has been a dramatic increase in scientific work supporting the concept that there are clinical benefits to ingesting specific nonpathogenic organisms (probiotics). The potential benefits of modifying the intestinal flora composition of certain high risk groups, eg, premature infants, travelers, and children receiving antibiotics, are emerging in the literature. Studies documenting prophylactic and therapeutic benefits in acute viral gastroenteritis and in atopic disease point not only to the potential applications, but also to the fact that the mechanisms of action of these agents may be due to their interaction with the gut as an immunologic organ. The benefits documented thus far are of varying degree and are most likely dependent on the number of agents, the dose, the dosing patterns, and the characteristics of the host and its underlying luminal microbial environment. Consequently, the safety and specification of a particular probiotic agent and methods of delivery to a particular population for a particular purpose should be carefully documented before making broad recommendations. The cost-benefit assessment of adding probiotics to our diet for prophylactic or therapeutic purposes, as well as better regulation of these agents as commercial products, is also needed. PMID- 11393194 TI - Probiotics: future directions. AB - Clinical studies have shown that certain probiotics may be useful in treating a variety of diarrheal disorders, including rotavirus diarrhea, antibiotic associated diarrhea, Clostridium difficile diarrhea, and traveler's diarrhea. New data suggest that probiotics might be useful in controlling inflammatory diseases, treating and preventing allergic diseases, preventing cancer, and stimulating the immune system, which may reduce the incidence of respiratory disease. Different modes of administering probiotics are currently being investigated, which may ultimately lead to the widespread use of probiotics in functional foods. It is important that such practices be directed by carefully controlled clinical studies published in peer-reviewed journals. PMID- 11393195 TI - Leak-proof puncture of ovarian cysts: instant mounting of plastic bag using cyanoacrylate adhesive. AB - BACKGROUND: Reliable leak-proof aspiration of cyst contents is required for treatment of large ovarian cysts by minilaparotomy. TECHNIQUE: Through a small abdominal wound a transparent plastic bag was instantly mounted onto the cyst surface using an ethyl-2-cyanoacrylate adhesive. A 1-2-cm-wide cut was made in the consolidated cyst wall through the inside of the bag and the contents directly aspirated. The fluid was trapped inside the bag without leaking into the abdominal cavity. This method can also be applied to relatively small cysts by holding the cyst just beneath the wound. EXPERIENCE: We used this method in 30 patients with unilateral ovarian cysts and in one patient with an ovarian cyst associated with an ipsilateral paraovarian cyst. All patients were successfully treated without spillage, although in one case a large mucinous ovarian cyst ruptured before surgery. CONCLUSION: Minilaparotomy using the instant adhesive is cost-effective, safe, reliable, and easily implemented. This procedure is also applicable to relatively small cysts and is a viable alternative to laparoscopic surgery for treatment of dermoid cysts showing considerable calcification. PMID- 11393196 TI - Laparoscopic suture hysteropexy for uterine prolapse. AB - OBJECTIVE: Vaginal hysterectomy remains the accepted surgical treatment for women with uterine prolapse. The Manchester repair is favored in women wishing uterine preservation. Vaginal hysterectomy alone fails to address the pathologic cause of the uterine prolapse. The Manchester repair has a high failure rate and may cause difficulty sampling the cervix and uterus in the future. The laparoscopic suture hysteropexy offers physiologic repair of uterine prolapse. METHOD: At the laparoscopic suture hysteropexy, the pouch of Douglas is closed and the uterosacral ligaments are plicated and reattached to the cervix. RESULTS: Forty three women with symptomatic uterine prolapse were prospectively evaluated and underwent laparoscopic suture hysteropexy with a mean follow-up of 12 +/- 7 months (range 6-32). The mean operating time for the laparoscopic suture hysteropexy alone was 42 +/- 15 minutes (range 22-121), and the mean blood loss was less than 50 mL. On review, 35 women (81%) had no symptoms of prolapse and 34 (79%) had no objective evidence of uterine prolapse. Two women subsequently completed term pregnancies and were without prolapse. Both underwent elective cesarean delivery. CONCLUSION: The laparoscopic suture hysteropexy is effective and safe in the management of symptomatic uterine prolapse. The result is physiologically correct, without disfiguring the cervix. This may be an appropriate procedure for women with uterine prolapse wishing uterine preservation. PMID- 11393197 TI - Umbilical artery N-terminal peptide of proatrial natriuretic peptide in hypertensive pregnancies and fetal acidemia during labor. PMID- 11393198 TI - Sexual activity during late pregnancy and risk of preterm delivery. PMID- 11393199 TI - Mode of delivery and risk od respiratory diseases in newborns. PMID- 11393200 TI - Unusual and memorable. Rheumatoid discitis. PMID- 11393201 TI - A comparison of somatotopic organization in sensory neocortex of newborn kittens and adult cats. PMID- 11393202 TI - Fluorescein dermofluorometry for the assessment of diabetic microvascular disease. AB - BACKGROUND/AIMS: Fluorescein dermofluorometry can be used to relate the uptake of fluorescein in the skin to blood flow. We have characterized the uptake of the dye by a wash-in time constant that is inversely proportional to the local blood flow. The purpose of this study was to explore the use of dermofluorometry in the assessment of patients with diabetic microvascular disease. METHODS: Fluorescein dermofluorometry was performed in four groups of patients: non-diabetic control patients, diabetic control patients, diabetic patients with chronic foot ulcers, and diabetic patients with acute foot ulcers. The outcomes of the patients with foot ulcers were documented 4-14 months after participation. Following an intravenous injection of sodium fluorescein, the change in the fluorescein signal with time was continuously measured at the plantar surface of the foot. Both the initial slope of the signal and the wash-in time constant were calculated in each subject. RESULTS: Significant differences in the wash-in time constant were found between diabetic and non-diabetic subjects and between diabetic subjects with and without foot ulcers. Of the eight patients with foot ulcers, two of them did not display an early wash-out in the dermofluorometer signal and later both required amputations. CONCLUSION: The fluorescein wash-in time constant demonstrated better correlation with the presence of diabetic microvascular disease than did the initial slope of the signal. Differences in the wash-in time constants of non diabetic and diabetic subjects support the hemodynamic hypothesis for the development of microvascular disease. The indication of early wash-out of the fluorescein signal may also be useful in the prediction of ulcer healing. PMID- 11393203 TI - Skin topography measurement by interference fringe projection: a technical validation. AB - BACKGROUND/AIMS: The quantitative analysis of skin topography is frequently used in cosmetology to evaluate the efficacy of hydrating or anti-wrinkle creams (micro-topography studies) or creams for slimming or to alleviate stretch marks (macro-topography studies). Numerous methods involving the three-dimensional (3 D) reconstruction of the topography from silicone replicas have been developed. Some of the main techniques applied include optical profilometry based on the reflection of oblique lighting or on transparency, and profilometry by laser focusing or triangulation. METHODS: We chose to test the Dermatop system developed by Eotech Co., which is based on interference fringe projection profilometry associated with the Toposurf surface processing software. Owing to the system's principle of operation, we first ran three experiments to evaluate the influence of ambient lighting on the 3-D reconstruction and the surface roughness parameters, the influence of the calibration device built into the acquisition system and the influence of calibration in terms of ambient lighting correction. We then evaluated the accuracy, the repeatability and the reproducibility of the measurements provided by the system from known metal topographic standards and skin replicas. RESULTS: It was seen that external lighting can cause variations in the calculation of parameters SPa, SPt and SPtm which can reach error levels of about 5% if the operator does not almost systematically calibrate the apparatus between each measurement. These three experiments enabled us to standardise the protocol for the acquisition of 3-D information, in order to minimise the lighting problems and yet respect the requirements for routine studies. The correlation coefficient between the theoretical values and the mean of experimental values was 0.9955 for parameter SPa and 0.9983 for parameter SPtm. Repeatability presented variations of under 4%, irrespective of the parameters, and reproducibility of under 6%. CONCLUSIONS: The technique provides very satisfactory results from the point of view of accuracy, repeatability and reproducibility. It is an excellent compromise between cost, accuracy and the time required. PMID- 11393204 TI - Measurement of the mechanical properties of skin with ballistometer and suction cup. AB - BACKGROUND: There is no consensus on the methodology for describing the mechanical properties of human skin in vivo. Current descriptions are generally method dependent. METHOD: The mechanical properties of palmar skin of the hypothenar as well as dorsal and ventral forearm skin were studied in 17 healthy volunteers. Two methods were used: ballistometry [Dia-stron Torsional Ballistometer (Diastron Ltd., Andover, UK)] and suction cup [The Dermaflex machine (Cortex Technology, Hadsund, Denmark)]. RESULTS: A moderate degree of correlation was found between the methods (rs=0.315-0.540), while internal correlation between different measures obtained with one method was higher (rs=0.375-0.967). The suction cup method parameters (distensibility and elasticity) correlated significantly with the ballistometry parameters (indentation, alpha, area and coefficient of restitution), while the hysteresis did not correlate to ballistometry parameters. The coefficient of variation of both methods (CV=0.02-0.35) was within the range obtained with other non-invasive methods, e.g., TEWL. Regional differences were identified with both methods, while only the suction cup method identified age-related changes. CONCLUSIONS: The results suggest that while both methods may be useful, they describe related but not identical aspects of skin mechanics. The differences in measuring principle suggest that the suction cup method predominantly measures elasticity, while the ballistometer predominantly appears to measure stiffness. Hysteresis may be a unique measure of skin viscosity. Additional studies, however, are needed to specify the clinical significance of the various measures of skin mechanics. PMID- 11393205 TI - Mechanical properties of the skin under suction. PMID- 11393206 TI - Measuring skin mechanics. The soft touch in a hard environment? PMID- 11393207 TI - Corticosteroid-induced atrophy and barrier impairment measured by non-invasive methods in human skin. AB - BACKGROUND/AIMS: Atrophy is a distressing side effect of potent corticosteroids. After open application of a high potency steroid, we monitored atrophogenicity by a variety of non-invasive methods. METHODS: Volar forearms were treated twice daily for 3 or 4 weeks, with clobetasol propionate cream (Temovate). The following methods were used: 1) confocal microscopy, 2) transepidermal water loss (TEWL), 3) dimethyl sulfoxide whealing, 4) sodium hydroxide erosions, 5) analysis of stratum corneum lipids, and 6) B-scan ultrasound. RESULTS: Confocal microscopy revealed thinning of the epidermis, decreased microvasculature and decreased size of keratinocytes. Evaporimetry demonstrated transepidermal water loss. Whealing to dimethyl sulfoxide was enhanced. Sodium hydroxide erosions formed more quickly. The amount of ceramides, cholesterol, and free fatty acids was reduced. Ultrasound showed thinning of the dermis. CONCLUSION: Non-invasive methods are very useful for quantifying the atrophogenicity of topical corticosteroids. PMID- 11393208 TI - Hand and forearm skin: comparison of their respective responsiveness to surfactants. AB - BACKGROUND/AIMS: Skin compatibility of detergent products is usually evaluated using predictive tests where products are applied on the back or the forearm of the volunteers, even if those products come more readily into contact with consumers' hands. The current study aimed at comparing the skin responsiveness of hands and volar and dorsal forearms to a surfactant solution. METHOD: Volunteers soaked simultaneously their forearms and hands in a solution of anionic surfactants, at 40 degrees C for 20 min, thrice a day and for 2 consecutive days. Skin condition on the hands, and the volar and dorsal forearms was evaluated by clinical examination and instrumental measurements. RESULTS: Significant differences were observed at baseline for various skin parameters, especially those related to skin dryness. After repetitive contacts with the surfactant solution, a parallel evolution of most skin parameters was observed--i.e., an absence of change in instrumentally measured erythema and transepidermal water loss, a similar decrease in corneometry values, and a similar increase in staining of the stratum corneum strippings and in clinical dryness. However, an opposite effect of the soakings on the cohesiveness between corneocytes harvested by tape strippings from hand and forearm was observed. CONCLUSION: For most parameters, skin responsiveness to surfactants appears to be quite similar on the hands and forearms. It is inferred that forearms can be conveniently used to predict interaction of surfactants with hands. PMID- 11393209 TI - Comparison of two pH meters used for skin surface pH measurement: the pH meter 'pH900' from Courage & Khazaka versus the pH meter '1140' from Mettler Toledo. AB - BACKGROUND/AIMS: Measurement of skin surface pH is used in clinical research to evaluate hazardous shifts in pH following external exposures and to evaluate the state of diseased skin with acute or chronic changes. It is therefore important to measure skin surface pH as precisely as possible. The aim of this study was to compare two commercially available pH meters used for skin surface pH measurement, to reveal differences between them in measured skin pH on the forearm. METHODS: The first pH meter (pH900) had a pointed electrode and a stabilisation period of 3 s. The second pH meter (pH meter 1140) had a circular electrode and no fixed stabilisation period. Twelve healthy subjects (6 male and 6 female Caucasians) entered the study. The pH measurements were performed once an hour from 8 a.m. to 3 p.m. on both forearms in five areas from the elbow to the "wristwatch" zone. In each area, three measurements were performed next to each other with both pH meters (15 measurements per arm per hour per pH meter). RESULTS: The pH900 has a higher measuring level and a higher variation than the pH meter 1140. CONCLUSION: A skin surface pH meter with a circular electrode and with no fixed stabilisation period is preferable. It is recommended that the pH meter be allowed to stabilise for at least 7 s before the result is read. PMID- 11393210 TI - Females have lower skin surface pH than men. A study on the surface of gender, forearm site variation, right/left difference and time of the day on the skin surface pH. AB - BACKGROUND/AIMS: It is of great interest to describe the many functions and properties of the skin in order to better understand reactions that result in different skin abnormalities, as a prerequisite in the development of skin products and topical medicines. Skin surface pH is considered a critical parameter of skin wellbeing and is typically studied on the forearm skin. Despite many previous investigations, this is an extensive field that still needs a great deal of research. The aim of our study was to investigate gender related differences, differences between right and left arms, anatomical variation and daytime variation in skin surface pH. METHODS: Skin surface pH was measured on the flexor surface of the forearm on eleven healthy volunteers (6 men and 5 women). A Mettler Toledo pH meter (pH meter 1140) was used. The subjects were measured once every hour from 8 a.m. to 3 p.m. in five areas from the elbow to the wrist. In each of the five areas, three measurements were performed next to each other, and the mean of these was used in subsequent calculations. RESULTS: A statistically significant difference in skin pH between men (mean pH=5.80) and women (mean pH=5.54) was found, with women being more acidic than men (P<0.01). No difference between right and left arm was found. In men, the area closest to the wrist had significantly lower pH values compared with the proximal sites. This was not the case in women. Skin surface pH decreased during normal working hours in both genders. CONCLUSION: Spontaneous skin surface pH was found to be significantly lower in women, as compared to men--albeit, the difference was small and of unknown relevance. Nevertheless, comparative studies on skin surface pH should be balanced with respect to gender. There appeared to be no right/left difference and no systematic change during the working day. Also, measurements should not be conducted close to the wrist. PMID- 11393211 TI - The use of high-frequency diagnostic ultrasound to investigate the effect of hormone replacement therapy on skin thickness. AB - BACKGROUND/AIMS: Previous investigations have suggested that hormone replacement therapy (HRT) could have a positive effect on the maintenance of skin thickness post-menopause. Previous skin measurement devices have proved variable in their accuracy and ease of use. This investigation assessed the effect of HRT on the skin in a noninvasive way, using high-frequency diagnostic ultrasound. METHOD: The study was a cross-sectional observational study, carried out at a menopause and gynaecology outpatient's clinic. A total of 84 women (comprising 34 HRT users, 25 post-menopausal controls, and 25 premenopausal controls) took part in the study. Each volunteer was scanned using diagnostic ultrasound on the arm. Skin thickness measurements were made from each scan using computerised image analysis. RESULTS: Skin thickness was shown to be greater in the HRT group than in the post-menopausal controls (P<0.01). CONCLUSIONS: High-frequency diagnostic ultrasound proved to be a useful clinical tool and showed that HRT appears to help maintain skin thickness in menopause. PMID- 11393212 TI - A comparative study of the effects on the skin of a classical bar soap and a syndet cleansing bar in normal use conditions and in the soap chamber test. AB - BACKGROUND/AIMS: The skin irritation potential of a body cleansing product is often compared under exaggerated test conditions, although the product is intended to be used at home with repetitive and brief contact with the skin. The aim of this study was to determine how much patch testing is predictive of the clinical, sub-clinical and subjective cutaneous effects of products used at home by consumers for their normal hygienic cleansing. METHODS: A double-blind comparative study of the normal use of an alkaline soap bar and a syndet at home during 10 consecutive weeks was performed on two identical groups of 25 healthy female subjects. The eventual skin changes observed at different anatomical skin sites were evaluated by clinical visual examination and by bioengineering measurements before the start of the study and then every 2 weeks. The objective measurements were compared with the subject's perceptions of dryness, tightness and product irritancy during the testing. RESULTS: The bioengineering measurements did not show any significant changes on all the anatomical skin sites, except for a small increase in skin pH with the classical soap bar. However, a trend appeared, showing that the alkaline soap bar is perceived by the subjects themselves as more of an irritant than the syndet bar. In the soap chamber test, the bar soap showed a significantly higher irritancy than the syndet bar. CONCLUSION: This study showed that cutaneous irritation induced by cleansing products in patch testing is not necessarily predictive of the irritation likely to occur in normal use conditions. Finally, a clear relationship could be demonstrated between the results of the soap chamber test and the consumer perception of both cleansing bars. PMID- 11393213 TI - Effects of holding time during cooling and of type of package on plasma membrane integrity, motility and in vitro oocyte penetration ability of frozen-thawed boar spermatozoa. AB - The effect of a prolonged holding time (HT) during cooling on plasma membrane integrity (PMI), motility and in vitro oocyte penetration ability of boar spermatozoa frozen-thawed in different types of package was investigated. Boar semen was frozen in a split-sample design using 3 different HTs (3, 10 and 20 h) during cooling and three different types of freezing package: Maxi-straws, Medium straws and FlatPacks. Assessment of PMI (SYBR-14 and propidium iodide, fluorescence microscopy) and sperm motility (visually and with CASA) was done during cooling (at 32 degrees C, 15 degrees C, 5 degrees C) and post-thaw (PT). The in vitro oocyte penetration ability of the spermatozoa was tested only PT, using a homologous in vitro penetration assay (hIVP). During cooling the HTs used had no significant (p<0.05) effect on either PMI or percentage of motile spermatozoa Post-thaw PMI was significantly higher (p<0.05) for 10 h and 20 h HT compared with 3 h, and the percentage of motile spermatozoa decreased significantly with 20 h HT as opposed to 3 h and 10 h. Regarding the freezing packages, the FlatPacks and Maxi-straws yielded significantly more PMI than did the Medium-straws (p<0.05). Post-thaw motility was significantly higher for FlatPacks than for straws, in terms of both percentage motile spermatozoa, and sperm velocity and lateral head displacement (LHD). The hIVP did not show any significant differences among the HTs, although FlatPacks yielded a significantly higher penetration rate and more spermatozoa per penetrated oocyte (p<0.05) than did the straws. Changes in motility patterns, toward a more circular motility during cooling and PT, could be noticed where individual spermatozoa showed a capacitation-like motility pattern. The changes were more obvious with 10-h and 20-h HTs than with 3-h HT. PMID- 11393214 TI - Effects of storage time and temperature on atresia of goat ovarian preantral follicles held in M199 with or without indole-3-acetic acid supplementation. AB - Maintenance of follicular quality after removal and during transport of ovaries is necessary for studies on development of preantral follicles in vitro. The present work investigated the effectiveness of M199 and M199IAA for preservation of goat preantral follicles in ovarian tissue. At the slaughterhouse, the ovarian pair of each animal was divided into 19 fragments. One ovarian fragment was immediately fixed (control--Time 0). The other 18 fragments were randomly distributed in M199 or M1991AA at 4, 20 or 39 degrees C and stored for 4, 12 or 24 h. Histological analysis showed that storage of ovarian fragments in either solution at 20 or 39 degrees C significantly reduced the percentage of normal preantral follicles when compared with the control, in all cases except after preservation in M199IAA at 20 degrees C for 4 h. In contrast, preservation at 4 degrees C, in either solution, kept the percentage of normal preantral follicles at control values. Reduced cellular metabolism may explain why the best preservation of preantral follicles was at 4 degrees C. The addition of IAA to the TCM 199 was effective for goat preantral follicle preservation at 20 degrees C for 4 h. PMID- 11393215 TI - The effect of the gonadotropin-releasing hormone analog, buserelin, on pregnancy rates in horse and pony mares. AB - We conducted a series of trials over a four-year period on a total of 2,346 mares, to determine the effect of a single dose of the GnRH analog buserelin (20 to 40 microg i.m. or s.c.) on pregnancy rates when given between 8 and 12 days after service. Although there were some statistically significant improvements in pregnancy rates in individual trials, meta-analysis of the data overall showed significant improvements at all times examined, i.e. 13 to 16, 19 to 23, 28 to 31 and 38 to 42 days after service. These results indicate that treatment of mares with 20 to 40 microg buserelin between Days 8 and 12 significantly increases pregnancy rates by approximately 10 percentage points. PMID- 11393216 TI - Photosensitive chemical and laser light treatments decrease epizootic hemorrhagic disease virus associated with in vitro produced bovine embryos. AB - Photoinactivation was employed to eliminate EHDV-2 from in vitro produced bovine embryos experimentally exposed to this virus. Immature oocytes were matured, fertilized, and cultured in chemically defined conditions. All treatments were performed on zygotes. Developmental potential of zygotes and cell numbers of resulting hatched blastocysts were assessed after exposure to a 1 mW helium neon laser (633 nm, red) for 1, 5, 10, and 15 min; the photosensitive chemicals hematoporphyrin (15 microM) and hypericin (1 and 10 microM) for 15 min; a combination of 10 microM hypericin and laser light for 1, 3, or 5 min; and a combination of 15 microM hematoporphyrin and laser light for 1, 2, or 3 min. There were no significant differences among proportions of embryos developing or cell numbers after treatment with or without exposure to laser light alone for up to 10 min. No differences were observed after exposure of zygotes to photosensitive chemicals alone. Exposure to 10 microM hypericin and 5 min of laser light or 15 microM hematoporphyrin and 2 min of laser light compromised zygote developmental potential. After exposure to 10(6) TCID50/mL EHDV-2 for 90 min groups of 10 zygotes were exposed to 10 microM hypericin or 15 microM hematoporphyrin and laser light to inactivate the virus. Hematoporphyrin was effective with 3 min light exposure at reducing the percentage of EHDV-2 contaminated zygote pools (16.7%) as compared to EHDV-2 exposed pools without treatment (88.9%) but hematoporphyrin + 1 min light was ineffective. Hypericin + 3 min light provided an intermediate effect (55.6%). PMID- 11393217 TI - Fertile estrus induced in bitches by bromocryptine, a dopamine agonist: a clinical trial. AB - The dopamine agonist bromocryptine, probably through amplifying gonadotroph (mainly FSH) secretion, was found to be suitable for provoking fertile estrus during the anestrous phase in bitches without functional cycles and/or ovarian activity. We studied estrus induction in 48 bitches after treatment with semisynthetic ergot alkaloid bromocryptine. For habituation a fractional dose of 0.3 mg/bitch was administered for three days followed by larger doses within the range of 0.6 to 2.5 mg/bitch by selecting dose rates on the basis of individual responsiveness and body weight. The long-term daily bromocryptine dose did not exceed 0.6 mg/bitch and 2.5 mg/bitch in small and large sized bitches, respectively. Gradual habituation and individual dose rates have almost completely eliminated the unwanted side effect of emesis. The period between treatment and onset of estrus varied but the average was 19 days. After the onset of estrus bromocryptine administration was usually continued for another 3 to 6 days. Occurrences of estrus, ovulation and pregnancy were monitored by cytological evaluation of vaginal epithelium, rapid ELISA for plasma progesterone and ultrasonography, respectively. Samples for progesterone were taken on Days 7, 9, 12 and 15 and sonograms of ovarian follicles and of fetuses were taken on Days 0, 22 and 35. The bitches involved in the study either regular or irregular cycles. Bromocryptine treatment induced estrus in all of the bitches including 40 of 48 (83%) with ovulation within a regular estrus and 6 of 48 (12.5%) that showed estrus but did not ovulate. Mating or artificial insemination of bitches in their fertile periods twice at two day intervals resulted in an 83% pregnancy rate (40 cases) and 39 (97.5%) of them gave birth to puppies. However, the average litter size was small with 4.8 +/- 1.6 pups. PMID- 11393218 TI - Effects of tumor necrosis factor-alpha on secretion of prostaglandins E2 and F2alpha in bovine endometrium throughout the estrous cycle. AB - To determine the physiological significance of tumor necrosis factor-alpha (TNFalpha) in the regulation of endometrial prostaglandin (PG) release in cattle, we investigated the effects of TNFalpha on the secretion of PGE2 and PGF2alpha by bovine endometrium during the estrous cycle. Bovine uteri were classified into six stages (estrus: Day 0, early luteal 1: Days 2 to 3, early luteal 11: Days 5 to 6, mid-luteal: Days 8 to 12, late luteal: Days 15 to 17 and follicular: Days 19 to 21). After 1 h of pre-incubation, endometrial tissues (20 to 30 mg) were exposed to 0 or 0.6 nM TNFalpha for 4 h. The PGE2 concentrations in the medium were higher in the luteal stages than in the follicular stage and in estrus. In contrast, PGF2alpha concentrations were higher in the follicular stage and in estrus than in the luteal stages. The ratio of the basal concentrations of PGE2 and PGF2alpha (PGE2/PGF2alpha ratio) was higher in the luteal stages than in the follicular stage and in estrus. Although TNFalpha stimulated both PGE2 and PGF2alpha secretion during the entire period of the estrous cycle, the level of stimulation of TNFalpha on PGE2 output by the bovine endometrium does not show the same cyclical changes as that shown on PGF2alpha output. The stimulation of TNFalpha resulted in a decrease in the PGE2/PGF2alpha ratio only in the late luteal stage. Furthermore, TNFalpha stimulated PGE2 secretion in stromal, but not epithelial cells. The overall results suggest that TNFalpha is a potent regulator of endometrial PGE2 secretion as well as PGF2alpha secretion during the entire period of estrous cycle, and that TNFalpha plays different roles in the regulation of secretory function of bovine endometrium at different phases of the estrous cycle. PMID- 11393220 TI - A separate procedure of fusion and activation in an ear fibroblast nuclear transfer program improves preimplantation development of bovine reconstituted oocytes. AB - This study was conducted to examine whether preimplantation development of bovine ("HanWoo," Bos Taurus corcanae) oocytes reconstituted with ear fibroblasts could be improved by a modified procedure of fusion and activation. In Experiment 1, enucleated oocytes were reconstituted with ear fibroblast by a combined procedure of electric fusion and activation at either 24 or 28 hours after IVM. The 28 hours reconstitution yielded more blastocysts (4% vs. 21%, P = 0.0025) and higher ratio of blastocysts per 2-cell embryos (0.05 vs. 0.25, P = 0.003) than the 24 hours. In Experiment 2, enucleated oocytes were reconstituted by one of the three fusion and activation protocols; 1) a combined procedure of electric fusion and activation at 28 hours after IVM, 2) a combined procedure of electric fusion and chemical activation at 28 hours, and 3) a separate procedure of electric fusion at 24 hours and chemical activation at 28 hours. When compared two combined procedures, chemical activation with 5 microM ionomycin for 4 minutes did not promote embryo development and significantly reduced the fusion rate (42% vs. 53%, P = 0.0395). However, significant (P < 0.0113) increases in the development to 2-cell (90% vs. 70 to 74%) and blastocyst (47% vs. 7 to 13%) stages and in the ratio of blastocysts per 2-cell embryos (0.52 vs. 0.11 to 0.18) were obtained by a separate procedure of electric fusion and chemical activation than by the combined procedures. This separate protocol did not reduce the fusion rate compared with the combined procedures (58%). In conclusion, improved development of oocytes reconstituted with ear fibroblasts was achieved by undergoing a separate procedure of electric fusion and chemical activation 4 hours apart. PMID- 11393219 TI - Use of buserelin to induce ovulation in the cyclic mare. AB - Inducing ovulation in a cyclic mare is often necessary. For this purpose, hCG has been used commonly, but the response can be reduced after successive administrations. The aims of this study were to test the effectiveness of buserelin in hastening ovulation in estrus mares, and its influence on fertility; and to investigate the effect of treatment on LH secretion. Five crossover trials were designed to compare the effect of two treatments: buserelin (40 microg in 4 doses i.v. at 12 h intervals) vs placebo (Experiments 1 and 2); buserelin 40 microg (in 4 doses i.v.) vs 20 microg (Experiment 3); buserelin (4 doses of 20 microg i.v.) vs hCG (1 dose of 2,500 IU i.v.) (Experiment 4); or buserelin (3 doses of 13.3 microg at 6 h interval) vs hCG (Experiment 5). In Experiment 2, blood samples were taken hourly until ovulation, for LH measurements. In Experiment 1, buserelin treatment significantly hastened ovulation. Reduction of the dose by half (Experiment 3) did not alter the effectiveness. In Experiments 4 and 5, buserelin was as effective as hCG in inducing ovulation between 24 and 48 h after initiation of treatment. Buserelin treatment induced a rise in LH concentration during the 48 h period of the experiment, and LH concentrations before ovulation were significantly higher in buserelin treated cycles than in placebo cycles. These experiments demonstrated the usefulness of two new protocols of administration of buserelin, as an alternative to hCG for induction of ovulation. One hypothesis explaining the mechanism of action is that the persistant rise in LH concentration could modify the ratio of biological/immunological LH, as it occurs physiologically, thereby hastening ovulation. PMID- 11393221 TI - Immunolocalization of zona pellucida antigens in the ovarian follicle of dogs, cats, horses and elephants. AB - A comparative evaluation of the location of immunoreactive porcine zona pellucida (pZP) glycoproteins was performed with polyclonal rabbit anti-pZP antibodies on ovarian sections of the dog, cat, horse, and elephant. For this, formalin (light microscopy) and glutaraldehyde (transmission electron microscopy [TEM]) fixed ovarian sections were incubated with antibodies raised against highly purified pZP. Staining patterns were determined with diaminobenzidine (DAB) at the light level. The dog ZP had a distinct staining distribution that is characterized by intense staining around the periphery of the ZP and the oolemma and less dense staining throughout the width of the ZP. In dog follicles that contained multiple oocytes, there were oocytes of identical and dissimilar stages. Cat ovarian sections showed uniform staining of the ZP. Horse results showed uniform staining of ZP and ooplasm, and granulosa cells (GC). Elephant sections showed staining of the ZP with dense staining at the oolemma, as well as staining of the ooplasm. In all species the staining of the ZP was not evident until GC differentiation. In all cases there was no staining of ovarian tissue with control normal rabbit serum. Specific staining patterns of ZP were evaluated by TEM and immunogold staining. The immunogold-linked anti-pZP antibodies stained the ZP matrix in all species. There was staining of ooplasm organelles suggesting that ZP secretion originates from the oocyte of the dog and cat. In addition, follicular and ZP measurements were taken that allowed accurate characterization of follicle stage. These findings suggest that in all four species the ZP is recognized by anti-pZP antibodies and there is also evidence to suggest the possible origins of ZP glycoproteins. PMID- 11393222 TI - Effect of cooling rate and partial removal of yolk on the chilling injury in zebrafish (Danio rerio) embryos. AB - High chilling sensitivity is one of the main obstacles to successful cryopreservation of zebrafish embryos. So far the nature of the chilling injury in fish embryos has not been clear. The aim of this study is to investigate the effect of cooling rate and partial removal of yolk on chilling injury in zebrafish embryos. Zebrafish embryos at 64-cell, 50%-epiboly, 6-somite and prim-6 stages were cooled to either 0 degrees C or -5 degrees C at three different cooling rates: slow (0.3 degrees C/min or 1 degree C/min), moderate (30 degrees C/min), and rapid (approximately 300 degrees C/min). After chilling, embryos were warmed in a 26 degrees C water bath, followed by 3-day culturing in EM at 26 +/- 1 degrees C for survival assessment. When embryos were cooled to 0 degrees C for up to 30 min, 64-cell embryos had higher survival after rapid cooling than when they were cooled at a slower rate. When 64-cell embryos were held at -5 degrees C for 1 min, their survival decreased greatly after both slow and rapid cooling. The effect of cooling rate on the survival of 50%-epiboly and 6-somite embryos was not significant after 1 h exposure at 0 degrees C and 1 min exposure at -5 degrees C. However, rapid cooling resulted in significantly lower embryo survival than a cooling rate of 30 degrees C/min or 1 degree C/min after 1 h exposure to 0 degrees C for prim-6 stage or 1 h exposure to -5 degrees C for all stages. Chilling injury in 64-cell embryos appears to be a consequence of exposure time at low temperatures rather than a consequence of rapid cooling. Results also indicate that chilling injury in later stage embryos (50%-epiboly, 6-somite and prim-6) is a consequence of the combination of rapid cooling and exposure time at low temperatures. Dechorionated prim-6 embryos were punctured and about half of yolk was removed. After 24 h culture at 26 +/- 1 degrees C after removal of yolk, the yolk-reduced embryos showed higher embryo survival than did control embryos after rapid cooling to -5 degrees C for 10 to 60 min. Results suggest that cold shock injury after rapid cooling can be mitigated after partial removal of yolk at the prim-6 stage. These findings help us to understand the nature of chilling sensitivity of fish embryos and to develop protocols for their cryopreservation. PMID- 11393223 TI - The influence of cAMP before or during bovine oocyte maturation on embryonic developmental competence. AB - This study was designed to evaluate the effects of pretreatment with various forms of cAMP before or during bovine oocyte maturation on the acquisition of embryonic developmental competence. The objective of the 4 experiments was to induce differentiation of the early maturing oocyte in conditions of maintained meiotic arrest or normal maturation. To promote differentiation, different forms of cyclic AMP-dependent protein kinase (PKA) pathways were investigated. The factors studied included follicular fluid, invasive adenylate cyclase (iAC), dibutyryl-cAMP (dbcAMP) and 3-isobutyl-1-methyl-xanthine (IBMX) with or without cycloheximide (CHX). High concentrations of iAC pretreatment were beneficial to the oocyte competence in BSA-iAC maturation while harmful in normal maturation. Also, after 2 to 3 h IBMX-iAC pretreatment, another 6 h of CHX treatment with or without iAC was harmful to the embryonic developmental competence of fertilized oocytes even though it did not have any effect on cleavage rate. Experiment 4 was to assess the role of cAMP in acquisition of oocyte developmental competence before meiotic resumption. Results supported that the intracellular cAMP concentration during the interval between oocyte isolation from the follicle and the beginning of in vitro maturation is critical for requiring optimal developmental competence. PMID- 11393224 TI - Association of Fusarium mycotoxicosis with failure in applying an induction of parturition program with PGF2alpha and oxytocin in sows. AB - This trial was conducted in a farrow-to-finish pig unit from November 1999 to February 2000. Since November 1998 an induction-of-parturition program was applied in gilts and sows with PGF2alpha (2 mL Dinolytic, i.m.) 113 d post service, followed by oxytocin (1 mL Intertocine-S, i.m.) 24 h later. This program resulted in a high proportion of animals farrowing within the working hours of the day. At mid December 1999 splay-legs and edematous swelling and reddening of the vulva started to be observed in newborn piglets. A concurrent decline of parameters related to parturition also was noticed. Mycotoxicological analyses of the feeds revealed a co-occurring contamination with deoxynivalenol and zearalenone. For a 4-week period, sows were divided into two groups: (a) an induction-of-parturition and (b) a non-induction-of-parturition group. Significant differences were found between the two groups relating to prevalence of dystocia (<.05) and pregnancy duration (<.05). Moreover, it was found that prevalence of splay-legs and swelling of the vulva were highly correlated (<.05) with reduction of percentage of sows farrowing within the working day and increase of pre-weaning mortality. It was concluded that such an induction-of parturition program should be avoided during a Fusarium mycotoxicosis. PMID- 11393225 TI - Short-term nonfrozen storage of mouse epididymal spermatozoa. AB - Six simple methods for short-term (up to 8 d), nonfrozen (5 to 20 degrees C) storage of mouse epididymides were compared with respect to the motility and fertility of spermatozoa. A high percentage of progressively motile spermatozoa was obtained from epididymis stored for 8 d at 5 degrees C in mineral oil (78.3%), covered with body fat (80.0%), or stored in the intact body of the euthanized donor animal (77.5%). Fertilized eggs (6.4% fertilization rate) were obtained by IVF using spermatozoa that had been stored in mineral oil at 5 degrees C for at least 8 d, and offspring were obtained from 77.5% of transferred eggs that were fertilized by spermatozoa stored for 2 d. These methods inhibited moisture loss from the preserved epididymal spermatozoa, thereby allowing spermatozoa to be stored for a few days without loss of either motility or fertility. These methods make possible such wide-ranging applications as the long distance transport of epididymis spermatozoa. While in storage at 5 degrees C, the tail of each recovered spermatozoon was bent midway along the tail, possibly owing to damage to the plasma membranes and due to the spermatozoa's hardening in the phospholipid by exposure to the low temperature. PMID- 11393226 TI - Cyclodextrins as electrode modifiers. AB - Complexation abilities and analytical applications of working electrodes with attached cyclodextrins (CDs) are reviewed. For the immobilization of CDs, their adsorption and formation of self-assembled monolayers, preparation of polymer films, as well as incorporation within plasticized membranes and composites such as carbon paste and gels are considered. Some electrochemical investigations of the CD interactions in solution are also reported with respect to their use at CD modified electrodes. PMID- 11393227 TI - Voltammetric determination of cadmium(II) using a chemically modified electrode. AB - An 1-(pyridylazo)-2-naphthol modified glassy carbon electrode has been investigated as sensor for the measurement of trace levels of Cd2+. Cd2+ is deposited on the surface of a PAN modified glassy carbon electrode at -1.10 V (vs. SCE) via forming Cd2+-PAN and subsequent reduction at the electrode. In the following step, Cd-PAN is oxidized, and voltammograms are recorded by scanning the potential in a positive direction. Calibration plots were found to be linear in the range 2 x 10(-8) mol/L to 8 x 10(-7) mol/L. The detection limit was 5 x 10(-10) mol/L, and the coefficient of variation, determined on one single electrode at a concentration of 5 x 10(-7) mol/L, was calculated to be 3.2% (n = 5). Using this new kind of modified electrode, trace levels of Cd(II) in water samples were determined; the average recovery was calculated to be 98.78%. PMID- 11393228 TI - A simple and rapid in situ preconcentration method for the determination of phosphate in environmental waters by use of solid-phase extraction, and its applications to brackish lake waters. AB - A simple and rapid in situ preconcentration method for the determination of phosphate in environmental waters has been developed for field analysis. This method is based on solid-phase extraction on a zirconium-loaded Sep-Pack Accell CM cartridge (Zr-SP) and is applicable to studies in which sampling is performed by use of a graduated syringe to prevent contamination and to ensure easy operation at sampling sites. The Zr-SP cartridge was prepared by passing 0.1 mol L(-1) zirconium solution through a Sep-Pak Accell CM cartridge, packed with cation exchange sorbent based on a silica matrix. The adsorption of phosphate and its desorption depend only on the pH of the solution. A water sample containing phosphate was adjusted to pH 2 and passed through the Zr-SP cartridge to collect it. The retained phosphate was quantitatively eluted with 0.5 mol L(-1) sodium hydroxide solution. The phosphate retained in the Zr-SP cartridge was stable for at least one month. The established preconcentration method was successfully applied to brackish lake waters to investigate seasonal changes in the distribution and behavior of phosphate in a brackish lake. PMID- 11393229 TI - Critical outlook and trends for environmental reference materials at the Measurements & Testing Generic Activity (European Commission). AB - It is now well recognised that the quality control (QC) of all types of analyses, including environmental analyses depends on the appropriate use of reference materials. One of the ways to check the accuracy of methods is based on the use of Certified Reference Materials (CRMs), whereas other types of (not certified) Reference Materials (RMs) are used for routine quality control (establishment of control charts) and interlaboratory testing (e.g. proficiency testing). The perception of these materials, in particular with respect to their production and use, differs widely according to various perspectives (e.g. RM producers, routine laboratories, researchers). This review discusses some critical aspects of RM use and production for the QC of environmental analyses and describes the new approach followed by the Measurements & Testing Generic Activity (European Commission) to tackle new research and production needs. PMID- 11393230 TI - A two-dimensional immune algorithm for resolution of overlapping two-way chromatograms. AB - A two-dimensional immune algorithm is proposed for resolving the multicomponent overlapping two-way data matrices. The method is a development of the one dimensional immune algorithm proposed elsewhere. When the inner product of vectors is expanded to the similar operation on matrices, the 1D immune algorithm can be expanded to the 2D algorithm which is suitable for the analysis of two-way data matrices. Both simulated and experimental two-way data sets were investigated by the method, and the results prove that the 2D immune algorithm is an effective tool for resolving the overlapping two-way signals. The effect of noise on the recoveries is also discussed. PMID- 11393231 TI - Flow-injection spectrophotometric multidetermination of metallic ions with a single reagent exploiting multicommutation and multidetection. AB - A flow system designed by computer-controlled discrete commutation devices is proposed for the multidetermination of metallic ions with a single chromogenic reagent. Intermittent addition of masking agents and changes of the reaction pH were exploited to implement selective determination of four species. Multidetection with an optical-fiber CCD-array spectrophotometer enabled simultaneous measurements at different wavelengths for multidetermination. The proposed system was applied to the determination of iron, copper, nickel and zinc in alloys and pharmaceutical preparations, yielding results in agreement with those obtained by flame atomic absorption spectrophotometry at the 95% confidence level. The sampling rate was estimated to be 80 determinations per hour. Coefficients of variation lower than 2% (n = 20) were achieved for all species. PMID- 11393232 TI - Example of a technique for evaluation of interferences caused by complicated sample matrix elements in ICP-AES determination. AB - An example of a useful and rapid procedure for the evaluation of interferences caused by complicated sample matrices in inductively coupled plasma atomic emission spectrometry (ICP-AES) is described. Using simple acid-base standards, all the elements investigated were determined separately in complicated matrices with satisfactory results. Multiple linear regression was used to calculate the linear correction coefficients for each matrix element analyzed. Good analytical results improved still further when this correction method was used. PMID- 11393233 TI - Maltodextrins as new chiral selectors in the design of potentiometric, enantioselective membrane electrodes. AB - Maltodextrins (dextrose equivalent (DE) 4.0-7.0, 13.0-17.0, and 16.5-19.5) are proposed as novel chiral selectors for the construction of potentiometric, enantioselective membrane electrodes. The potentiometric, enantioselective membrane electrodes can be used reliably for the assay of S-captopril as raw material and in pharmaceutical formulations such as Novocaptopril tablets, by use of direct potentiometry. The best response was obtained when maltodextrin with higher DE was used for construction of the electrode. The best enantioselectivity and time-stability was achieved for the lower DE maltodextrin. L-proline was found to be the main interferent for all the proposed electrodes. The surface of the electrodes can be regenerated by simply polishing; this furnishes a fresh surface ready for use in a new assay. PMID- 11393234 TI - The validation of Kayzero-assisted NAA in Budapest, Rez, and Ljubljana via the analysis of three BCR certified reference materials. AB - After installation and calibration of k0-assisted NAA in three Central European research institutes (AEKI-Budapest, NPI-Rez, and IJS, Ljubljana), its validation was established via the analysis of three BCR certified reference materials. The matrices of choice were: CRM 277 estuarine sediment, CRM 038 coal fly ash from pulverized coal, and CRM 101 spruce needles. For some elements, e.g. Zn, Cd, and Hg, the analyses were not only performed instrumentally (INAA), but also in the radiochemical mode (RNAA). The work was performed in the framework of a European Copernicus Project. PMID- 11393235 TI - The certification of SRM 1546--Meat Homogenate, a new reference material for nutrients in a high protein, high fat matrix. AB - In response to reference material needs expressed by the food industry and government regulators, the National Institute of Standards and Technology (NIST) has developed a new Standard Reference Material (SRM) consisting of a canned meat product with certified and reference values for a large number of constituents. SRM 1546 Meat Homogenate consists of a mixture of finely ground pork and chicken prepared and canned by a commercial process. NIST determined the concentration levels of cholesterol, sodium, calcium, iron, and seven fatty acids in this SRM using well defined methods and procedures. These analytes as well as 34 other constituents or properties were determined in an interlaboratory comparison exercise involving 21 laboratories, most of which are associated with the National Food Processors Association (NFPA) Food Industry Analytical Chemists Subcommittee (FIACS). From statistical analysis of the data, NIST assigned certified concentrations for the eleven analytes measured at NIST and reference concentrations for the proximates, six additional fatty acids, seven minerals, and seven water-soluble vitamins. Information values without uncertainties are provided for the concentrations of six additional constituents for which the uncertainties could not adequately be assessed. SRM 1546 will provide laboratories with a means to evaluate the accuracy of the methods they use to assign nutrient levels to processed meats and similar products. PMID- 11393236 TI - Determination of hydrogen ion by ion chromatography (IC) with sulfonated cation exchange resin as the stationary phase and aqueous EDTA (ethylenediamine N,N,N',N'-tetraacetic acid) solution as the mobile phase. AB - An ion chromatographic (IC) method has been developed for determination of hydrogen ion (H+). It is based on the use of sulfonated cation-exchange resin as stationary phase, aqueous ethylenediamine-N,N,N',N'-tetraacetic acid (dipotassium salt, EDTA-2K, written as K2H2Y) solution as mobile phase, and conductivity for detection. H+ was separated mainly by cation-exchange, but its elution was accelerated by the presence of EDTA. The order of elution for the model cations was H+ > Li+ > Na+ > NH4+ > Ca2+ > > Mg2+. A sharp and highly symmetrical peak was obtained for H+ and this was attributed to the capacity of H2Y2(2-) to receive and bind H+. H+ was detected conductiometrically and detector response (reduction in conductivity as a result of H+ +H2Y2- --> H3Y-) was linearly proportional to the concentration of H+ in the sample. The detection limit for H+ with this IC system was better than 4.7 micromol L(-1). A significant advantage of this method was the ability to separate and determine, in one step, H+ and other cations. The successful determination of H+ and other cation species in real acid-rain samples demonstrated the usefulness of this method. PMID- 11393237 TI - Application of high-surface-area ZrO2 in preconcentration and determination of 18 elements by on-line flow injection with inductively coupled plasma atomic emission spectrometry. AB - A flow-injection analysis (FIA) system incorporating a micro-column of ZrO2 has been used for the development of an on-line multi-element method for the simultaneous preconcentration and determination of Al, Bi, Cd, Co, Cr, Cu, Fe, Ga, In, Mn, Mo, Ni, Pb, Tl, V, Sb, Sn, and Zn by inductively coupled plasma atomic emission spectrometry (ICP-AES). The conditions for quantitative and reproducible preconcentration, elution, and subsequent on-line ICP-AES determination were established. A sample (pH 8) is pumped through the column at 3 mL min(-1) and sequentially eluted directly into the ICP-AES with 3 mol L(-1) HNO3. With a sample volume of 100 mL and an elution volume of 1 mL signal enhancement 100 times better than for conventional continuous aspirating systems was obtained for the elements studied. The reproducibility (RSD %) of the method at the 10 ng mL(-1) level in the eluate is acceptable - less than 8% for five replicates. Recoveries between 95.4% and 99.9% were obtained for the elements analysed. ZrO2, with a specific surface area of 57 m2 g(-1) and a capacity of approximately 5 mg g(-1) for the elements studied, was synthesized by hydrolysis of ZrCl4. The preconcentration system was evaluated for several simple synthetic matrices, standard water samples and synthetic seawater. The effect of foreign ions on the efficiency of preconcentration of the elements studied was investigated. The application of a micro-column filled with high-surface-area ZrO2 and flow injection inductively coupled plasma atomic emission spectrometry enables preconcentration and simultaneous determination of 18 elements at low concentrations (ng L(-1)) in different water samples. PMID- 11393238 TI - Determination of elemental sulfur in coal by gas chromatography-mass spectrometry. AB - A method for the determination of S0 in coal based on the extraction with cyclohexane with subsequent quantitative analysis of elemental sulfur in the extract by GC/MS is described. The quantity of elemental sulfur was determined in four coal samples with different distribution of sulfur forms. The effect of solvent and extraction time on the efficiency of sulfur removal was studied. The elemental sulfur extracted from coal occurred in the form of S6, S7 and S8. Calibration solutions were prepared from freshly recrystalized elemental sulfur. It was found that the injection temperature has a crucial influence on the m/z 64 ion chromatogram. PMID- 11393239 TI - Determination of trace amounts of lead in mussels by flow-injection flame atomic absorption spectrometry coupled with on-line minicolumn preconcentration. AB - A minicolumn packed with poly(aminophosphonic acid) chelating resin incorporated in an on-line preconcentration system for flame atomic-absorption spectrometry was used to determine ultratrace amounts of lead in mussel samples at microg L( 1) level. The preconcentrated lead was eluted with hydrochloric acid and injected directly into the nebulizer for atomization in an air-acetylene flame for measurement. The performance characteristics of the determination of lead were: preconcentration factor 26.8 for 1 min preconcentration time, detection limit (3sigma) in the sample digest was 0.25 microg g(-1) (dry weight) for a sample volume of 3.5 mL and 0.2 g sample (preconcentration time 1 min), precision (RSD) 2.3% for 25 microg L(-1) and 2.0% for 50 microg L(-1). The sampling frequency was 45 h(-1). The method was highly tolerant of interferences, and the results obtained for the determination of lead in a reference material testify to the applicability of the proposed procedure to the determination of lead at ultratrace level in biological materials such as mussel samples. PMID- 11393240 TI - Direct mass-quantification of particulate deposits of polycyclic aromatic hydrocarbons by Fourier transform imaging microscopy. AB - Successful coupling of imaging microscopy with Fourier transform spectrometry provides a new methodological approach. This was applied to the direct analysis of particulate deposits of polycyclic aromatic hydrocarbons (PAHs). The fluorescence signals of single microscopic particulates were found to be proportional to their mass, obtained from the corresponding HPLC results. Special fluorescence characteristics related to individual particle analysis were studied. The sensitivity of the proposed method to PAHs in soil is in the sub-ppb range, similar to HPLC results. Analysis of inhomogeneous PAH-particulates was demonstrated, as well as the resolution of photochemical products of PAHs. The effects of particulate orientation relative to the detector upon quantification are discussed. PMID- 11393241 TI - An evaluation of analytical techniques for determination of lead, cadmium, chromium, and mercury in food-packaging materials. AB - Closed microwave digestion and a high-pressure asher have been evaluated for wet oxidation and extraction of lead, cadmium, chromium, and mercury from a range of typical packaging materials used for food products. For the high-pressure asher a combination of nitric and sulfuric acids was efficient for destruction of a range of packaging materials; for polystyrene, however, nitric acid alone was more efficient. For microwave digestion, a reagent containing nitric acid, sulfuric acid, and hydrogen peroxide was used for all materials except polystyrene. Use of the high-pressure asher resulted in the highest recoveries of spiked lead (median 92%), cadmium (median 92%), chromium (median 97%), and mercury (median 83%). All samples were spiked before digestion with 40 microg L(-1) Cd, Cr, and Pb and 8 microg L(-1) Hg in solution. The use of indium as internal standard improved the accuracy of results from both ICP-MS and ICP-AES. Average recovery of the four elements from spiked packaging materials was 92 +/- 14% by ICP-MS and 87 +/- 15% (except for mercury) by ICP-AES. For mercury analysis by CVAAS, use of tin(II) chloride as reducing agent resulted in considerably better accuracy than use of sodium borohydride reagent. PMID- 11393242 TI - Analytical performance of luminescent immunoassays of different format for serum daidzein analysis. AB - Two sensitive competitive-type solid-phase immunoassays for serum daidzein analysis have been developed and optimized. The first is a chemiluminescent enzyme immunoassay that uses black polystyrene microtiter wells in which daidzein specific antibodies raised in rabbits are immobilized and a daidzein derivative is coupled to horseradish peroxidase (HRP) as a label. The HRP activity of the antibody-bound tracer is measured with an enhanced chemiluminescent system (luminol/H2O2/enhancer). The second immunoassay is based on the use of bovine serum albumin-daidzein derivative immobilized on microtiter plates and a secondary anti-rabbit IgG-Fc fragment conjugated with 4,7-bis(chlorosulfophenyl) 1,10-phenanthroline-2,9-dicarboxylic acid (BCPDA). Formation of the complex Eu3+ BCPDA enables time-resolved fluorescence-mode detection of the amount of antibody bound to the immobilized antigen. Both methods fulfilled all the requirements of accuracy and precision. The detection limit was the same for each method, 10 pg/well; this is better than that of other immunoassays. The specificity of the two methods was different, because of their competitive-type mechanisms. The performance of the chemiluminescence method is better, because the cross reactivity of the main interfering compound (genistein) was 5%, compared with 25% for the time-resolved fluoroimmunoassay. PMID- 11393243 TI - Fluorescence immunoassay of alpha-fetoprotein with iron(III) tetrasulfonatophthalocyanine as a mimetic enzyme labeling reagent. AB - A new fluorimetric immunoassay for alpha-fetoprotein (AFP) has been developed using a novel promising mimetic peroxidase, iron(III) tetrasulfonatophthalocyanine (FeTSPc), as a labeling reagent to catalyze the fluorescence reaction of P- hydroxyphenylacetic acid (P-HPA) and hydrogen peroxide (H2O2). In the competitive immunoassay, anti-AFP antibody was coated on a 96-well plate (polystyrene) and a constant amount of FeTSPc-labeled AFP and a known amount of test solution were added. Non-labeled and FeTSPc-labeled AFP compete for binding to the plate-bound antibody. After the immunoreaction, the immunochemically adsorbed FeTSPc-AFP conjugate moiety was determined by measuring the fluorescence produced in a solution containing P-HPA and H2O2. AFP can be determined in the concentration range of 1-300 ng mL(-1) with a detection limit of 0.5 ng mL(-1). PMID- 11393244 TI - HPLC-based method using sample pre-column clean-up for the determination of methanethiol and ethanethiol in parenteral amino acid solutions. AB - A method has been developed for the chromatographic determination of methanethiol (MT) and ethanethiol (ET) as contaminants in amino acid parenteral nutrition (PN) solutions. The clean-up of the samples before chromatographic analysis was investigated by solid-phase extraction (SPE) on pre-columns filled with polyethylene powder (PE), aluminium oxide (AlOx), silica (SiOx), or polyurethane foam (PUF) as adsorbents. The thiols were more efficiently separated from the matrices by SPE on PUF pre-columns. Simultaneous derivatization and elution with DTNB (5,5'-dithiobis(2-nitrobenzoic acid)) enabled further discrimination between MT and ET by reversed-phase HPLC with spectrophotometric detection. The retention times for the derivatized MT and ET species were 12.5 and 23.0 min, respectively. Recoveries from spiked PN samples were calculated to be approximately 90%, and the MT and ET content of commercial PN solutions was determined using the methodology described. Detection limits of 15 and 10 microg L(-1) were calculated for MT and ET, respectively. PMID- 11393245 TI - 2-Chloroacetaldehyde induces epsilondA DNA adducts in DNA of Raji cells as demonstrated by an improved HPLC-fluorimetry method. AB - Etheno adducts in DNA are promutagenic lesions and are formed from vinyl chloride, urethane, and lipid peroxidation-derived products such as 4 hydroxynonenal. Such adducts were induced in a B-lymphoid cell line (Raji) by a metabolite of vinyl chloride, 2-chloroacetaldehyde. By modification of a method of Bedell et al., etheno dA was determined in 20 microg DNA by HPLC-fluorimetry. Our method has a detection limit of 5 fmol which is one two-hundredth that of the original method. By use of this method initial evidence was found that cigarette smoke can also induce etheno dA adducts in Raji cell DNA. PMID- 11393246 TI - Developing a new model for cross-cultural research: synthesizing the Health Belief Model and the Theory of Reasoned Action. AB - This article discusses the development of a new model representing the synthesis of two models that are often used to study health behaviors: the Health Belief Model and the Theory of Reasoned Action. The new model was developed as the theoretic framework for an investigation of the factors affecting participation by Mexican migrant workers in tuberculosis screening. Development of the synthesized model evolved from the concern that models used to investigate health seeking behaviors of mainstream Anglo groups in the United States might not be appropriate for studying migrant workers or persons from other cultural backgrounds. PMID- 11393247 TI - Meaningfulness as an epistemologic concept for explicating the researcher's constitutive part in phenomenologic research. AB - In his philosophy, Husserl gave us the epistemologic concept of intentionality. In phenomenologic research, intentionality can be understood in a practical way as our connections to the phenomena that we are studying. When phenomenologic data are the transcripts of interviews, our connections can be seen during data analysis as we identify passages in transcripts that stand out as personally meaningful. Tracing back through our past experiences to the origins of that meaningfulness provides us with a picture of the preunderstanding, assumptions, and beliefs that contribute to our unique perception of the phenomena that we are studying. Concrete guidelines are given for initiating the process of discovering one's constitutive part in phenomenologic studies. PMID- 11393248 TI - Unitary appreciative inquiry. AB - Unitary appreciative inquiry is described as an orientation, process, and approach for illuminating the wholeness, uniqueness, and essence that are the pattern of human life. It was designed to bring the concepts, assumptions, and perspectives of the science of unitary human beings into reality as a mode of inquiry. Unitary appreciative inquiry provides a way of giving fullest attention to important facets of human life that often are not fully accounted for in current methods that have a heavier emphasis on diagnostic representations. The participatory, synoptic, and transformative qualities of the unitary appreciative process are explicated. The critical dimensions of nursing knowledge development expressed in dialectics of the general and the particular, action and theory, stories and numbers, sense and soul, aesthetics and empirics, and interpretation and emancipation are considered in the context of the unitary appreciative stance. Issues of legitimacy of knowledge and credibility of research are posed and examined in the context of four quality standards that are deemed important to evaluate the worthiness of unitary appreciative inquiry for the advancement of nursing science and practice. PMID- 11393249 TI - Methodology for social accountability: multiple methods and feminist, poststructural, psychoanalytic discourse analysis. AB - Bridging the gap between the individual and social context, methodology that aims to surface and explore the regulatory function of discourse on subjectivity production moves nursing research beyond the individual level in order to theorize social context and its influence on health and well-being. This article describes the feminist, poststructural, psychoanalytic discourse analysis and multiple methods used in a recent study exploring links between cultural discourses of masculinity, performativity of masculinity, and practices of male violence. PMID- 11393250 TI - A feminist critique on the use of the Internet in nursing research. AB - With the increasing use of the Internet, the importance of incorporating this new technology in nursing research increases. Yet nursing has been slow in adopting this new technology as a research method, and the use of the Internet in nursing research rarely has been discussed and critiqued. In this article, use of the Internet in nursing research is analyzed and critiqued from a feminist perspective. The analysis indicates that use of the Internet in research frequently brings about selection biases because of a limited pool of potential participants, it usually does not provide contextual data on research encounters, it might not uncover women's subjective experiences under marginalized situations, and it tends to raise power issues related to the relationships between researchers and participants. Despite the limitations, use of the Internet in research provides better communication channels for research participants, more flexibility in time and place of data collection, and less expense for data collection. Based on the critique, feminist challenges for future use of the Internet in nursing research are proposed. PMID- 11393251 TI - Markers for cell-mediated immune response are elevated in cerebrospinal fluid and serum after severe traumatic brain injury in humans. AB - The brain is believed to be an immunologically privileged organ, sheltered from the systemic immunological defense by the blood-brain barrier (BBB). However, there is increasing evidence for a marked inflammatory response in the brain after traumatic brain injury (TBI). Markers for cellular immune activation, neopterin, beta2-microglobulin (beta2M), and soluble interleukin-2 receptor (sIL 2R), were measured for up to 3 weeks in cerebrospinal fluid (CSF) and serum of 41 patients with severe TBI in order to elucidate the time course and the origin of the cellular immune response following TBI. Neopterin gradually increased during the first posttraumatic week in both CSF and serum. Concentrations in CSF were generally higher than in serum, suggesting intrathecal release of this marker. beta2M showed similar kinetics but with higher serum than CSF concentrations. Nonetheless, intrathecal release as assessed by the beta2M index could be postulated for most of the patients. The mean levels of sIL-2R in both CSF and serum were elevated during the whole study period, serum concentrations being up to 2 x 10(4) times higher than in CSF. No significant intrathecal production of sIL-2R could be detected. The present data shows that severe TBI leads to a marked cell-mediated immune response within the brain and in the systemic circulation. In the intrathecal compartment the activated cells appear to be predominantly of the macrophage/microglia lineage, while the immune activation in the systemic circulation seems to involve mainly T-lymphocytes. PMID- 11393252 TI - S-100 beta reflects the extent of injury and outcome, whereas neuronal specific enolase is a better indicator of neuroinflammation in patients with severe traumatic brain injury. AB - It has been hypothesized that immunoactivation may contribute to brain damage and affect outcome after traumatic brain injury (TBI). In order to determine the role of inflammation after TBI, we studied the interrelationship of the immune mediators sICAM-1 and IL-6 with the levels of S-100beta and neuronal specific enolase (NSE), both recognized markers of brain damage. In addition, the extent and type of cerebral injury and the neurological outcome were related to these measured markers of injury. An evident elevation of S-100beta (range of means: 2.7-81.4 ng/mL) and NSE (range of means: 2.0-81.3 ng/mL) was observed in CSF of all 13 patients during the first 3 posttraumatic days and decreased over 2 weeks. In parallel, the production of sICAM-1 (range of means: 0.7-11.9 ng/mL) and IL-6 (range of means: 0.1-8.2 ng/mL) was also markedly enhanced in CSF. The CSF means of S-100beta and NSE per patient correlated with IL-6 (r = 0.60, p < 0.05; and r = 0.64, p < 0.05, respectively), whereas the corresponding means in serum showed a significant correlation only between NSE and IL-6 (r = 0.56, p < 0.05). Maximal CSF values of NSE and sICAM-1 correlated with each other (r = 0.57, p < 0.05). The contusion sizes assessed on the CT scans correlated with the means of S 100beta (r = 0.63, p < 0.05) and NSE (r = 0.71, p < 0.05) in CSF and with the mean of S-100beta in serum, although not statistically significant (r = 0.52, p = 0.06), but not with serum NSE. Interestingly, linear regression analysis demonstrated that means of S-100beta in CSF (r = 0.78, p = 0.002) and serum (r = 0.82, p < 0.001) correlated with the GOS. These results indicate that the elevation of these parameters in CSF depends on the extent of injury and that S 100beta may be a predictor of outcome after TBI, whereas NSE reflects better the inflammatory response. PMID- 11393253 TI - Dynamic stretch correlates to both morphological abnormalities and electrophysiological impairment in a model of traumatic axonal injury. AB - In this investigation, the relationships between stretch and both morphological and electrophysiological signs of axonal injury were examined in the guinea pig optic nerve stretch model. Additionally, the relationship between axonal morphology and electrophysiological impairment was assessed. Axonal injury was produced in vivo by elongating the guinea pig optic nerve between 0 and 8 mm (Ntotal = 70). Morphological damage was detected using neurofilament immunohistochemistry (SMI 32). Electrophysiological impairment was determined using changes in visual evoked potentials (VEPs) measured prior to injury, every 5 min for 40 min following injury, and at sacrifice (72 h). All nerves subjected to ocular displacements greater than 6 mm demonstrated axonal swellings and retraction bulbs, while nerves subjected to displacements below 4 mm did not show any signs of morphological injury. Planned comparisons of latency shifts of the N35 peak in the VEPs showed that ocular displacements greater than 5 mm produced electrophysiological impairment that was significantly different from sham animals. Logit analysis demonstrated that less stretch was required to elicit electrophysiological changes (5.5 mm) than morphological signs of damage (6.8 mm). Moreover, Student t tests indicated that the mean latency shift measured in animals exhibiting morphological injury was significantly greater than that calculated from animals lacking morphological injury (p < 0.01). These data show that distinct mechanical thresholds exist for both morphological and electrophysiological damage to the white matter. In a larger context, the distinct injury thresholds presented in the report will aid in the biomechanical assessment of animate models of head injury, as well as assist in extending these findings to predict the conditions that cause white matter injury in humans. PMID- 11393254 TI - Cyclosporin A treatment following spinal cord injury to the rat: behavioral effects and stereological assessment of tissue sparing. AB - The immunosuppressant drug cyclosporin A (CsA) has significant neuroprotective properties following CNS injury. In the present study, we assessed the efficacy of CsA therapy following a moderate spinal cord injury (SCI). Adult female rats were injured with the NYU impactor from a height of 12.5 mm, and CsA or vehicle therapy was initiated 15 min after the injury. All animals were behaviorally tested with the BBB locomotor rating scale prior to morphological assessment of changes in the spinal cord. CsA therapy failed to significantly improve the behavioral recovery following the injury. Using a unique stereological approach to assess tissue damage, it was determined that CsA did not alter the amount of spared tissue. The possible neuroprotective effects of CsA, observed in other models of CNS injury, do not appear to influence SCI pathology, perhaps reflecting both anatomical and physiological differences between these distinct regions of the CNS. PMID- 11393255 TI - iNOS and nitrotyrosine expression after spinal cord injury. AB - Secondary tissue damage after spinal cord injury (SCI) may be due to inflammatory mediators. After SCI, the nuclear factor-kappaB (NF-kappaB) transcription factor can activate many pro-inflammatory genes, one of which is inducible nitric oxide synthase (iNOS). iNOS catalyzes the synthesis of nitric oxide (NO), a key inflammatory mediator, which in turn reacts with superoxide to generate peroxynitrite. Peroxynitrite is a strong oxidant that can damage cellular enzymes, membranes, and subcellular organelles through the nitration of tyrosine residues on proteins. The presence of nitrotyrosine (NT) is an indirect chemical indicator of toxic NO and peroxynitrite-induced cellular damage. Using a New York University (NYU) impactor to induce SCI in adult rats, we examined the temporal and cellular expression of iNOS and NT. We observed a progressive increase in iNOS expression in the injured cord starting at day 1 with maximal expression occurring at day 7, as determined by Western blot analysis. iNOS expression corresponded temporally to an increase in iNOS enzyme activity after SCI. In parallel with the progressive increase in iNOS activity, NT expression also increased with time after SCI. The iNOS and NT immunoreactivity was localized in neurons, astrocytes, endothelial cells and ependymal cells at the epicenter and adjacent to the region of spinal cord impact and injury. Results from the present study suggest that increased iNOS and peroxynitrite anion, as reflected by the progressive accumulation of NT in the injured impacted spinal cord, may contribute to the secondary injury process after SCI. PMID- 11393256 TI - Methylprednisolone reduces spinal cord injury in rats without affecting tumor necrosis factor-alpha production. AB - Methylprednisolone (MPS) is the only therapeutic agent currently available for traumatic spinal cord injury (SCI). However, little is known about its therapeutic mechanisms. We have demonstrated that tumor necrosis factor-alpha (TNF-alpha) plays a critical role in posttraumatic SCI in rats. Since MPS has been shown to inhibit TNF-alpha production in vitro, it is possible that MPS can reduce SCI by inhibiting TNF-alpha production. To examine this possibility, we investigated the effect of MPS on TNF-alpha production in injured segments of rat spinal cord. Leukocytopenia and high-dose intravenous administration of MPS markedly reduced the motor disturbances observed following spinal cord trauma. Both treatments also reduced the intramedullary hemorrhages observed histologically 24 hr posttrauma. Leukocytopenia significantly reduced tissue levels of both TNF-alpha mRNA and TNF-alpha, 1 and 4 hr posttrauma, respectively, and it also inhibited the accumulation of leukocytes in the injured segments 3 hr posttrauma, while MPS had no effects. Lipid peroxidation and vascular permeability at the site of spinal cord lesion were both significantly increased over time after the induction of SCI, peaking 3 hr posttrauma. These events were significantly reduced in animals with leukocytopenia and in those given anti-P selectin monoclonal antibody compared to sham-operated animals. Administration of MPS significantly inhibited both the increase in lipid peroxidation and the vascular permeability. These findings suggested that MPS reduces the severity of SCI, not by inhibiting the production of TNF-alpha at the site of spinal cord trauma, but by inhibiting activated leukocyte induced lipid peroxidation of the endothelial cell membrane. This suggests that MPS may attenuate spinal cord ischemia by inhibiting the increase in endothelial permeability at the site of spinal cord injury. PMID- 11393257 TI - Studies on the mechanisms responsible for the formation of focal swellings on neuronal processes using a novel in vitro model of axonal injury. AB - A novel in vitro model of axonal injury using PC12 cells was designed to introduce traumatic alterations on neuronal processes and to identify mechanisms responsible for the formation of focal swellings by observation with phase contrast and transmission electron microscopes. The injury on the processes was produced by one-dimensional, horizontal oscillation. The fluid shear stress applied by the oscillation did not exceed 380 dyne/cm2. The injured processes showed two forms. One involved an increase in the terminal diameter of the processes and the other entailed beading along the injured portions. Long-term observation of cellular responses to the mechanical insult disclosed that the terminal swelling coincided with the detachment of growth cones from the culture plate. The finding suggests that the detachment of the growth cone destroys the cytoskeletal network, which determines and maintains the cell shape, resulting in spherical deformation of the processes. When the cytoskeletal destruction occurred at non-terminal sites along the processes, spherical deformations developed slowly, and these appeared as beads. The beading also caused the detachment of the growth cones. As the most proximal bead grew, they absorbed the distal segment and their growth cones were pulled proximally with the spreading cytoskeletal destruction. The processes with terminal swellings as well as the bead segments showed regeneration with time evidence of and growth cone formation. PMID- 11393258 TI - Effects of the immunomodulator linomide on macrophage migration and myelin phagocytic activity in peripheral nerve trauma: an experimental study. AB - Wallerian degeneration after peripheral nerve transection leads to the phagocytosis of degenerated myelin and axon components by macrophages. These phagocytes are recruited from the systemic circulation and Wallerian degeneration may therefore be used as a model for myelin removal by hematogenous macrophages, a feature that is also a hallmark of demyelinating diseases of the central and peripheral nervous system. The immunomodulator linomide has been shown to be effective in the treatment of experimental demyelinating diseases although the exact mode of its action is not yet defined. The present study investigated the effect of linomide on monocyte invasion and myelin phagocytosis after sciatic nerve transection. Linomide had a dual effect in Wallerian degeneration. Monocyte migration from the circulation to the damaged nervous system was significantly reduced. Additionally, the myelin phagocytic capacity of macrophages was impaired, finally resulting in a significant delay in the removal of myelin. The present experiments may provide an explanation for the effects of linomide during the course of demyelinating diseases of the nervous system. PMID- 11393259 TI - Cellular localization of tumor necrosis factor-alpha following acute spinal cord injury in adult rats. AB - Posttraumatic inflammatory reaction may contribute to secondary injury after traumatic spinal cord injury (SCI). Expression of tumor necrosis factor-alpha (TNF-alpha), a key inflammatory mediator, has been demonstrated in the injured cord. However, the specific cell types that are responsible for TNF-alpha expression after SCI remain to be identified. In the present study, cellular sources of TNF-alpha were examined in rats that received a spinal cord impact injury at the 9th thoracic (T9) level. Here we demonstrate that, within hours after SCI, increased TNF-alpha immunoreactivity was localized in neurons, glial cells (including astrocytes, oligodendrocytes, and microglia), and endothelial cells in areas of the spinal cord adjacent to the lesion site. Myelin breakdown was noted in oligodendrocytes that are immunopositive for TNF-alpha. In sham operated controls, a low level of TNF-alpha immunoreactivity was detected. In antigen-absorption experiments, no TNF-alpha immunoreactivity was detected, indicating the specificity of TNF-alpha immunocytochemistry in the present study. Results suggest that various cell types, including neurons, glial cells, and vascular endothelial cells, contribute to TNF-alpha production in the injured cord. PMID- 11393260 TI - Neuron-specific enolase serum levels after controlled cortical impact injury in the rat. AB - The aim of this study was to investigate the time course and the correlation of neuron-specific enolase (NSE) serum levels to the severity of traumatic brain injury in rats. Sixty-five male Wistar rats were subjected to severe cortical impact injury (100 PSI, 2 mm deformation). Blood samples were drawn directly after trauma and after 1, 6, 12, 24, and 48 h in the trauma group. In the sham operated levels animals samples were drawn directly after craniotomy and after 6 and 48 h. Additionally, NSE serum levels after controlled cortical impact at different levels of severity samples (45 PSI, 75 PSI; 2 mm deformity) were compared to sham-operated animals. The severity of the injury was not validated histopathologically. NSE serum levels were estimated with a commercially available enzyme immunoassay (LIA mat Sangtec). The control animals showed a NSE serum level of 8.82 microg/L (mean, n = 10) and the injured animals demonstrated a time-dependent release of NSE into the serum. The highest NSE serum values were detected 6 h after trauma (31.5 microg/L mean, n = 10). In addition, we found a close relationship between NSE serum levels and the severity of traumatic brain injury in the cortical impact model. NSE serum levels reflect in a time-dependent manner the severity of brain trauma induced by cortical impact model in rat. PMID- 11393261 TI - The role of chemotherapy and prophylactic bilateral oophorectomy in a case of colorectal adenocarcinoma with ovarian metastases. AB - A 66-year-old female presented with a large abdominal mass and accompanying systemic complaints of abdominal pain, constipation. and fever. On exploratory laparotomy, the mass was found to be a moderately differentiated adenocarcinoma of the sigmoid colon with metastasis to the left ovary. A primary colorectal carcinoma that has metastasized to the ovaries can be difficult to distinguish clinically from an advanced primary ovarian tumor. Histology and tumor markers are currently the most useful tools available in making an accurate diagnosis. If the nature of the primary tumor is uncertain and the initial response to chemotherapy is poor, the patient's prognosis will also he poor. Though controversy exists regarding the role of prophylactic bilateral oophorectomy during resection for primary colorectal cancer, later confusion can be avoided by performing this procedure when the colorectal carcinoma is first diagnosed. However the possibility of a concurrent primary ovarian tumor must not be overlooked. PMID- 11393262 TI - The role of laparoscopy in the diagnosis and treatment of peritoneal carcinomatosis: a case report. AB - A patient presented with deep venous thrombosis and an elevated CA-125 level, but normal pelvic ultrasound and abdominal and pelvic CT scans. Laparoscopy revealed diffuse carcinomatosis and a diagnosis of stage IIIc, poorly differentiated epithelial ovarian carcinoma was made. Laparoscopy may provide an alternative means of diagnosis when conventional imaging fails, and may facilitate the placement of catheters for subsequent intraperitoneal therapy. PMID- 11393263 TI - Surgery at Yale in 1843: Professor Jonathan Knight's lectures. PMID- 11393264 TI - Activities of daily living and cardiovascular complications following elective, noncardiac surgery. AB - BACKGROUND: Algorithms for preoperative cardiac evaluation prior to noncardiac surgery use indices of the metabolic equivalent of activities of daily living (METs). We evaluated METs as a predictor of cardiac complications following elective, noncardiac surgery. METHODS: A study was performed in an outpatient university preadmission center METs were estimated prospectively for 5,939 inpatients admitted for elective, noncardiac surgery who underwent a preanesthetic assessment within two months prior to surgery. Cardiac outcomes were retrieved retrospectively from relational databases. Outcomes included death, myocardial infarction, acute congestive failure, arrhythmias, cardiac arrest, acute ischemia, acute renalfailure, stroke, respiratory failure, severe hypertension, peripheral vascular occlusion, and pericardial effusion. Adverse outcomes were correlated with age, gender, surgical procedure, activities, and the American Society of Anesthesiologist's Physical Status (ASA-PS) using receiver operator characteristic curve analysis. RESULTS: 94 of 5,939 (1.6 percent) patients had cardiac complications; 16 died, six from their cardiac complication. 38.3 percent of complications occurred following vascular surgery. Using a multinomial logistic regression analysis, both age and physical status were highly significant predictors (p < 0.001) but METs was not (p = 0. 793). Receiver operator characteristic (ROC) curves were usedfor predictive value of variables. Area of the curves for age versus cardiac complications and death were 0.814 and 0.782; for physical status, 0.744 and 0.803; for METs, 0.664 and 0.524. CONCLUSIONS: METs are not a reliable index for the prediction of adverse cardiac events following elective, noncardiac surgery. Age and physical status are more predictive. Adverse cardiac outcomes are most frequent following vascular surgery. PMID- 11393265 TI - Hypertension and its related factors in Taiwanese elderly people. AB - BACKGROUND: Our study used data collected in Chung-Hsing Village in May 1998 to evaluate the prevalence of hypertension and its correlates in Taiwanese elderly people. METHODS: All of individuals aged 65 and over were recruited as study subjects. A total of 1,093 persons, out of 1,774 registered residents, were contacted by face-to-face interview The response rate was 61.6 percent. However, only 586 respondents had blood tests and completed questionnaires. Analysis in this study was based on these 586 subjects. In order to study the significant correlates of hypertension, the t-test, chi-square analysis, and multivariate logistic regression were used. RESULTS: Our results showed that 66 percent were men and 34 percent were women. The mean age was 73.1 +/- 5.3 years. The proportions of hypertension were 53.09 percent in men and 56.06 percent in women (p > 0.05). After controlling the other covariates, the multivariate logistic regression analysis showed that the significant related factors of hypertension were obesity (OR = 1.88, 95 percent CI = 1.06-3.34, p < 0.05) and renalfunction impairment (OR = 1.69, 95 percent CI = 1.02-2.80, p < 0.05). CONCLUSIONS: The prevalence of hypertension was high in elderly people. Hypertension is significantly associated with obesity and renalfunction impairment in elderly people. PMID- 11393266 TI - Topical hyperbaric oxygen and low energy laser therapy for chronic diabetic foot ulcers resistant to conventional treatment. AB - Chronic foot ulcers are common in long-standing diabetes, may herald severe complications and are often resistant to therapy. To evaluate the effects of adjunctive topical hyperbaric oxygen treatment (THBO) and low energy laser (LEL) irradiation on ulcer healing, a 100 consecutive patients with chronic diabetic foot ulcers (DFU) refractory to 4.5 +/- 1.2 months of comprehensive treatment, were enrolled in a prospective open study. While conventional treatment was continued as necessary, THBO was administered by pumping 100 percent oxygen into a disposable sealed polythylene hyperbaric chamber (150 min x 2 to 3/wk at up to 1.04 atm). Helium-neon LEL irradiation was given concurrently using a Unilaser Scan Unit at 4 J/cm2 for 20 min. Some patients continued THBO at home or their treatment was confined to THBO at home. Patients were monitored every two weeks revealing 81 percent cure after 25 +/- 13 treatments over 3.2 +/- 1.7 months. On follow-up (median 18 months), only 3/81 (4 percent) had reulceration, which responded to THBO/LEL retreatment. Nonresponders had significantly lower ankle brachial indices (ABI) than patients whose ulcers were healed (0.55 vs. 0.78, p < 0.01) and ultimately required amputation. Patient compliance was full and no adverse events occurred. In conclusion, although the study was open and uncontrolled, an 81 percent healing of DFU in patients who previously did not respond to a comprehensive treatment program, constitutes an intriguing preliminary result. Thus, THBO/LEL therapy may be a safe, simple, and inexpensive early adjunctive treatment for patients with chronic diabetic foot ulcers. Our findings should prompt its evaluation by large randomized controlled trials. PMID- 11393267 TI - Neuroparalysis and oxime efficacy in organophosphate poisoning: a study of butyrylcholinesterase. AB - The temporal profile of butyrylcholinesterase (BuChE) and in vitro pralidoxime reactivated BuChE was studied in a cohort of 25 organophosphate-poisoned patients to examine their relationship to the development of intermediate syndrome and to understand reasons for lack of efficacy of oxime treatment. The clinical severity of poisoning (assessed by the Namba Scale) correlated significantly with the severity of intermediate syndrome. BuChE activity increased significantly over time and showed significant relationship to muscle power. The temporal profile of the enzyme was correlated to the clinical severity of poisoning. Reactivation potentials of BuChE (the difference between oxime-reactivated and -unreactivated enzyme activity) declined significantly with time after organophosphate ingestion. The reactivation potential of the enzyme at admission decreased significantly with increasing severity of poisoning and was lower in patients who developed intermediate syndrome. Patients who received oxime prior to hospitalization had a higher rate of intermediate syndrome and lower levels of BuChE at admission than those who had not. The study suggests that (i) BuChE reflects the clinical course of poisoning, confirming earlier studies; (ii) intermediate syndrome may be associated with a persistent inhibition of BuChE; and (iii) the lack of oxime efficacy in our patients maybe due to their severity of poisoning and the timing of oxime treatment. PMID- 11393268 TI - Carbamazepine poisoning: treatment with plasma exchange. AB - Lethal cases due to carbamazepine overdose have been reported. There are contradicting reports about the efficiency of hemodialysis, hemoperfusion and plasmapheresis for the treatment of carbamazepine poisoning. We present a case of carbamazepine intoxication successfully managed with plasma exchange. The patient was a 15-year-old girl. On admission there was no evidence of trauma, Glascow Coma Scale scored 6. Further questioning of the parents revealed the patient had taken at least 23 tablets of Tegretol (4.6 g) 6 h before the admission. The carbamazepine level was 190 micromol/l. Orogastric lavage was followed by activated charcoal. Within 20 h after admission there was no improvement in her neurological status. It was thus decided to perform plasmapheresis. At the end of the procedure she started to respond to verbal stimuli. Carbamazepine level immediately after the procedure was 101 micromol/l, and at the 36th, 60th and 84th hours were 72, 33 and 20 micromol/l, respectively. The patient was discharged on the fourth day. We have not observed any rebound in our patient. Thus we suggest that simple plasma exchange by plasma replacement is a cheaper and effective method for the treatment of intoxication with carbamazepine or similar drugs. PMID- 11393269 TI - Determination of coumarin metabolism in Turkish population. AB - Cytochrome P450 2A6 is an important human hepatic P450 which activates precarcinogens and oxidizes some drug constituents such as coumarin, halothane, and the major nicotine C-oxidase. Genetic polymorphism exists in the CYP2A6 gene. CYP2A6*1 (wild type) is responsible for the 7-hydroxylation of coumarin. The point mutation (T to A) in codon 160 leads to a single amino acid substitution (Leu to His) and the resulting protein, CYP2A*2 is unable to 7-hydroxylate coumarin. Gene conversion in exons 3, 6, and 8 between the CYP2A6 and the CYP2A7 genes creates another variant, CYP2A6*3. In this study, healthy male and female Turkish volunteers (n = 50) were administered 2 mg coumarin, and urine samples were analyzed for their content of the coumarin metabolite, 7-hydroxycoumarin (7OHC), by high-performance liquid chromatography (HPLC). Genetic polymorphism for CYP2A6 was detected by using two-step polymerase chain reaction (PCR) to identify CYP2A6*1, CYP2A6*2, and CYP2A6*3 in 13 of these subjects. The percentage of the dose excreted of total 7OHC in relation to CYP2A6 genotype and excretion of nicotine/cotinine was also evaluated to demonstrate the role of CYP2A6 in nicotine metabolism. The majority of Turkish subjects (68%) excreted less than 60% of the 2-mg dose as coumarin metabolite. The allelic frequencies were detected as 0.88 for CYP2A6*1 allele; 0.12 for CYP2A6*3 allele in 13 individuals. No heterozygous and homozygous individuals were identified for the CYP2A6*2 allelic variant. Phenotyping and genotyping for drug metabolizing enzymes are of great importance in studies correlating precarcinogen activation or drug metabolism to the CYP2A6 genotype in smoking behavior when populations are investigated. PMID- 11393270 TI - Analysis of single hair by XRF discloses mercury intake. AB - Utilizing information obtained from the X-ray fluorescence linear scanning along a single strand of hair in a recent fatal exposure to dimethylmercury, it was possible to determine the circumstances leading to the fatal result. When the Dartmouth chemistry professor displayed symptoms of mercury toxicity, samples of her urine and blood were found to have considerable amounts of mercury by a commercial laboratory. The Dartmouth medical people immediately started treatment with a chelator on January 29, 1997 to remove the mercury, and sent the first samples of blood, urine and hair taken on January 31, 1997 to our laboratory. Our X-ray fluorescence analysis of a single strand of hair shown in Figure 1 shows a single large peak of intake of mercury confirming the information revealed by examination of her laboratory notebook that she had spilled some dimethylmercury about 5 months previous to the date the hair sample was taken. Figure 2 shows two peaks, the peak closest to the scalp end of the hair shows the effect of the large amount of mercury released by the chelator, part of which appeared in the blood. Utilizing the start of the first increase in the blood level as August 14, 1996, and the second as caused by the chelator on January 29, 1997, a count of the number of 2-mm points measured between the two dates gives an accurate growth of the hair during that time. A close examination of Figure 1 indicates that it required five points, each of 2 mm, along the hair to reach the maximum. This indicates a time period of 10 mm of growth equal to 23 days during which a large concentration of methylmercury was entering the blood as evidenced by the hair concentration. The professor died on June 11, 1997. PMID- 11393271 TI - The management of Elephant's Ear poisoning. AB - The Araceae family of plants is the major cause of symptomatic plant ingestions in some developed countries (Dieffenbachia and Philodendron) and in Zimbabwe (Elephant's Ear), especially in children. A retrospective case series was carried out to evaluate the management of poisoning due to Elephant's Ear at the largest referral hospital in Zimbabwe for the period January 1995-December 1999. The study revealed inappropriate use of antibiotics, atropine, and antihistamines in the treatment of Elephant's Ear poisoning. This article also reviews the management of poisoning due to the Araceae family of plants as exemplified by Elephant's Ear. There is a need to educate health care workers on the clinical management of Elephant's Ear poisoning especially in developing countries where there are limited resources. PMID- 11393272 TI - Study of the toxicological effect of mitomycin C in mice: alteration on the biodistribution of radiopharmaceuticals used for renal evaluations. AB - Mitomycin C (MMC) has been used as a component of many chemotherapeutic regimens and some toxic effects of this substance have been reported. As it has been reported that the toxicological effect of a drug can alter the biodistribution of radiopharmaceuticals and because patients on chemotherapeutic treatment can be submitted to a nuclear medicine procedure, we investigated whether MMC could affect the uptake of various technetium-99m (99mTc) radiopharmaceuticals used for renal evaluations. The purpose of this study was to suggest a model to evaluate the toxic effect of substances in specific organs. Three doses of MMC (0.45 mg) were administered to mice (N=15). One hour after the last dose, 99mTc radiopharmaceuticals, 99mTc-diethylenetriaminepentaacetic acid (99mTc-DTPA), 99mTc-dimercaptosuccinic acid (99mTc-DMSA) or 99mTc-glucoheptonic acid (99mTc GHA), with activity of 7.4 MBq, were also administered in the treated group and in the control group (N=15). After another 0.5 h, the animals were sacrificed. The organs were isolated, the 99mTc radiopharmaceutical uptake in the organs quantified in a well counter and the percentages of radioactivity (%ATI) calculated. The results have shown that: (i) with 99mTc-DTPA, the %ATI increased in the pancreas, ovary, uterus, stomach, kidney, spleen, thymus, heart, lung, liver, thyroid and bone; (ii) with 99mTc-DMSA, the %ATI decreased in all the organs except for the brain; and (iii) with 99mTc-GHA, the %ATI increased in the liver and decreased in the stomach, thymus, heart and thyroid. The effects of this chemotherapeutic drug on the biodistribution of these radiopharmaceuticals were statistically significant (Wilcoxon test, p<0.05) and could be explained by the metabolization and/or therapeutic action of MMC. Studies with other radiopharmaceuticals are in progress. PMID- 11393273 TI - Effect of dimethyl sulfoxide on gentamicin-induced nephrotoxicity in rats. AB - Nephrotoxicity of gentamicin (GM) has been suggested to be mediated by the generation of reduced oxygen metabolites. The present study investigated the possible protective role of the free radical scavenger dimethyl sulfoxide (DMSO) on some indices of GM nephrotoxicity in rats. The antibiotic was injected intramuscularly (i.m.) at a dose of 100 mg/kg for six consecutive days, either with or without treatment with DMSO (12.5%, 25% or 50% in saline) at an intraperitoneal (i.p.) dose of 2 ml/kg 4 days before GM, and concomitantly with GM treatment thereafter. DMSO (25% in saline) was also given as above to rats treated with GM at i.m. doses of 25, 50 or 100 mg/kg for six consecutive days. GM caused dose-dependent significant increases in the concentrations of urea and creatinine in plasma, and in thiobarbituric acid reactive substances (TBARS) level in the kidney cortex and also caused significant decreases in the concentrations of reduced glutathione (GSH) and superoxide dismutase (SOD) activity. In GM-treated rats, DMSO dose-dependently lowered the elevated plasma urea and creatinine concentrations, and the rise in cortical TBARS. It also restored the levels of GSH and SOD activity to near normal. DMSO (25%) was effective in completely preventing the development of signs of nephrotoxicity of G (50 mg/kg). Treatment of the rats with DMSO alone, at any of the above doses, did not alter significantly any of the renal or hepatic function tests studied, and did not appear to adversely affect the kidney or liver histology. However, the efficacy and safety of DMSO require further studies. It is suggested that DMSO has potential protective effect against GM nephrotoxicity in rats. PMID- 11393274 TI - Changes in zinc and copper homeostasis in human livers and kidneys associated with exposure to environmental cadmium. AB - This study was undertaken to assess changes in zinc and copper homeostasis in human tissues that could be attributed to human exposure to environmental cadmium, using samples of lung, liver and kidney cortex of 61 Queensland residents, aged 2 to 89 years, who had died of accidental causes. None of the subjects were exposed to cadmium in the workplace. Levels of zinc in liver and kidney cortex samples showed inverse associations with donor age whereas zinc in lung only showed inverse association with gender. Lung zinc levels in females were 14% lower than in males. Zinc in liver and kidney cortex samples were found to exist in at least two pools; one was associated with cadmium that bound to metallothionein (MT) and the other was associated with non-MT bound copper. In liver, the amounts of zinc in the MT pool were smaller compared to those in non MT pool given that only 7% of zinc variations were explained by cadmium whereas 22% of the liver zinc variations were accounted for by non-MT bound copper. In sharp contrast, larger amounts of zinc in kidney cortex samples were in the MT pool, compared to those in the non-MT pool given that cadmium was found to explain 69% of total zinc variation whereas copper explained only 17% of kidney zinc variations. The levels of copper in liver were found to be increased by 45 50% in subjects with high cadmium exposure level, compared to subjects of similar ages with medium exposure level. The levels of zinc and copper in kidney cortex samples in the subjects with high cadmium exposure were both found to be significantly elevated compared to those found in the medium-exposure group whereas copper contents were about 19-23% greater than in medium- as well as low exposure groups. Taken together these results indicate increased sequestration of zinc and copper in liver and kidney cortex samples. The increases in metal sequestrations were observed in liver samples having cadmium contents of greater than 1 microg/g wet weight and in kidney cortex having cadmium contents of greater than 26 microg/g wet weight. Zinc and copper contents in lung of this sample group, however, were not associated with cadmium due probably to lower exposure levels compared to those of liver and kidney. PMID- 11393275 TI - Asymptomatic QTc prolongation associated with quetiapine fumarate overdose in a patient being treated with risperidone. AB - We report a patient who ingested a 2000-mg overdose of quetiapine fumarate (Seroquel). Her maintenance medications also included risperidone, venlafaxine, topiramate, and clonazepam. On presentation, she was drowsy, but had no other significant CNS signs and no cardiac symptoms or abnormal physical signs. Approximately 2 h after the quetiapine ingestion, an electrocardiogram (ECG) showed normal sinus rhythm at 95 beats/min with a corrected QT (QTc) interval of 537 ms (upper limit of normal = 440 ms). Plasma quetiapine concentration at that time was 1800 ng/ml. Continuous ECG monitoring for the subsequent 18 h did not reveal any episode of ventricular tachycardia. A 12-lead ECG 18 h post-overdose was normal with a QTc interval of 401 ms and the corresponding plasma quetiapine concentration was 160 ng/ml. She made an uneventful medical recovery from the toxic ingestion. This case suggests that when patients overdose on quetiapine while taking therapeutic doses of risperidone, such overdoses, even if not massive, can cause considerable QTc interval prolongation. We recommend that quetiapine overdose patients undergo continuous ECG monitoring for 12-18 h post ingestion. PMID- 11393276 TI - Additive effect of adenovirus-mediated E2F-1 gene transfer and topoisomerase II inhibitors on apoptosis in human osteosarcoma cells. AB - Recently, it has been demonstrated that Etoposide, a topoisomerase II inhibitor, can induce apoptosis in MDM2-overexpressing tumor cells by inhibition of MDM2 synthesis. We have previously shown that E2F-1 overexpression induces apoptosis of MDM2-overexpressing sarcoma cells, which is related to the inhibition of MDM2 expression. Therefore, the present study was designed to investigate the in vitro and in vivo effect of combined treatment of adenovirus-mediated E2F-1 and topoisomerase II inhibitors on the growth inhibition and apoptosis in human sarcoma cells. Two human sarcoma cell lines, OsACL and U2OS, were treated with topoisomerase II inhibitors (Etoposide and Adriamycin), alone or in combination with adenoviral vectors expressing beta-galactosidase (Ad-LacZ) or E2F-1 (Ad-E2F 1). E2F-1 expression was confirmed by Western blot analysis. Ad-E2F-1 gene transfer at a low dose (multiplicity of infection, 2) markedly increased the sensitivity of human sarcoma cells to topoisomerase II inhibitor treatment. This cooperative effect of E2F-1 and topoisomerase II inhibitors was less marked in SAOS-2 cells (p53 and pRb null). Topoisomerase II inhibitors also cooperated with E2F-1 overexpression to enhance tumor cell killing in an in vivo model using xenografts in nude mice. When combined with Adriamycin or Etoposide, E2F-1 adenovirus therapy resulted in approximately 95% and 85% decrease in tumor size, respectively, compared to controls (P<.05). These results suggest a new chemosensitization strategy that is effective in MDM2-overexpressing tumors and may have clinical utility. PMID- 11393277 TI - Preclinical and therapeutic utility of HVJ liposomes as a gene transfer vector for hepatocellular carcinoma using herpes simplex virus thymidine kinase. AB - Although gene therapy has been suggested to be a novel strategy to treat hepatocellular carcinoma (HCC), no study showing the clinical feasibility of vectors to treat HCC has been reported. In this preclinical study, we show evidence indicating that hemagglutinating virus of Japan (HVJ) liposomes are a feasible vector to treat HCC in a clinical setting using ganciclovir (GCV) and herpes simplex virus thymidine kinase (HSV-tk), which is driven by the cytomegalovirus immediate early enhancer/promoter (plasmid pcDNA3/HSV-tk). In in vitro experiments, almost complete tumor cell regression was achieved with the optimal GCV concentration (100 microg/mL) and more than 1/3 regression was seen even with a 20% transduction ratio using HuH7 HCC cells stably transformed by HSV tk. HVJ liposomes showed a 19.7% (mean) transduction rate of the lacZ gene in a relatively large mass of more than 300 mm3 in vivo, which is a clinically detectable size, implanted into SCID mice. Moreover, a single HSV-tk injection of HVJ liposomes followed by GCV treatment inhibited tumor growth at least within a week, and repeat administration was more effective. Furthermore, subcutaneous injection of an HVJ liposomes vehicle induced no apparent inflammatory response in C3H/HeN mice, whereas lacZ gene transfection resulted in inflammatory pathology, suggesting a lower immunogenicity of the HVJ envelope protein than those of bacteria-derived plasmid DNA or the beta-galactosidase gene product. From these findings, we conclude that HVJ liposomes are a clinically safe and effective gene transfer vector to treat HCC. PMID- 11393278 TI - DNA vaccination against neu reduces breast cancer incidence and metastasis in mice. AB - The gene for HER2/neu is overexpressed in 30-40% of breast and ovarian cancers, and this overexpression correlates with increased metastasis and poor prognosis. The HER2/neu gene product, a transmembrane protein kinase member of the EGF receptor family, has significant potential as a tumor antigen for vaccination. We inserted the sequence for neu into a novel plasmid called ELVIS containing a Sindbis virus replicon that reproduces multiple copies of mRNA. Mice vaccinated one time intramuscularly demonstrated a strong antibody response against A2L2, a murine breast cancer cell line transfected to express neu. Vaccinated mice challenged in the mammary fatpad with A2L2 had reduced tumor incidence and reduced tumor mass compared to mice challenged with tumor cells lacking the neu insert. Intradermal vaccination was also protective and required 80% less plasmid for a similar level of protection. Vaccination reduced the incidence of lung metastasis from mammary fatpad tumors and reduced the number of lung metastases resulting from intravenous injection of A2L2 cells. Cytotoxic T lymphocytes cultures of immune spleen cells with P815-neu cells produced high levels of interferon-gamma indicating an antigen-specific Th1-type immune response resulting from the vaccination. In a spontaneous breast tumor model using neu transgenic mice, vaccination with ELVIS-neu protected against development of spontaneous breast tumors. Our preclinical data indicate that therapeutic vaccination of patients with ELVIS-neu may reduce metastasis from HER2/neu expressing breast and ovarian tumors. PMID- 11393279 TI - A herpes simplex virus type 1 mutant deleted for gamma34.5 and LAT kills glioma cells in vitro and is inhibited for in vivo reactivation. AB - To create an oncolytic herpes simplex virus type 1 (HSV-1) that is inhibited for reactivation, we constructed a novel herpes recombinant virus with deletions in the gamma34.5 and LAT genes. The LAT gene was replaced by the gene for green fluorescent protein, thereby allowing viral infection to be followed. This virus, designated DM33, is effective in killing primary and established human glioma cell lines in culture. DM33 is considerably less virulent following intracerebral inoculation of HSV-susceptible BALB/c mice than the wild-type HSV-1 strain McKrae. The safety of this virus is further supported by the retention of its sensitivity to ganciclovir and its relatively limited toxicity against cultured human neuronal cells, astrocytes, and endothelial cells. The ability of DM33 to spontaneously reactivate was tested in a rabbit ocular infection model that accurately depicts human herpes infection and reactivation. Following ocular infection of rabbits, spontaneous reactivation was detected in 83% (15/18) of the eyes infected with wild-type McKrae. In contrast, none of the eyes infected with DM33 had detectable reactivation. The efficacy of this virus in cultured human glioma cell lines, its safety, confirmed by its inability to reactivate, and its attenuated neurovirulence make DM33 a promising oncolytic agent for tumor therapy. PMID- 11393280 TI - Antitumor effect of B16 melanoma cells genetically modified with the angiogenesis inhibitor rnasin. AB - The growth of new blood vessels is an essential condition for the development of tumors with a diameter greater than 1-2 mm and also for their metastatic dissemination. RNasin, the placental ribonuclease inhibitor, is known to have antiangiogenic activity through the inhibition of angiogenin and basic fibroblast growth factor. Nevertheless, the administration of the recombinant form of a protein poses several limitations; as a result, we have studied the antitumor effect of RNasin in a murine gene therapy model. RNasin cDNA was subcloned into the pcDNA3 expression vector, and the resulting recombinant plasmid was used to transfect the B16 murine melanoma cell line. An RNasin inverted construction was used as control. Mice intravenously injected with clones expressing RNasin showed a significant inhibition of tumor metastatic progression with respect to control groups (P<.001) and survived longer (P<.001). Tissue sections from RNasin expressing cell tumors showed a lower number of blood vessels when compared to tissue sections from mice lungs that had been inoculated with control cell lines. The results of these experiments show that the genetic modification of tumor cells with RNasin cDNA yields a significant antitumor effect, and suggest that this effect is at least partially the result of angiogenesis inhibition. PMID- 11393281 TI - Suppression of oncogenic viral interferon regulatory factor (vIRF) of Kaposi's sarcoma-associated herpesvirus by ribozyme-mediated cleavage. AB - Kaposi's sarcoma-associated herpesvirus/human herpesvirus 8 (KSHV/HHV8) has been etiologically associated with several malignancies including Kaposi's sarcoma and primary effusion lymphoma. Oncogenic viral interferon regulatory factor (vIRF) encoded by KSHV ORF-K9 is a homologue of cellular interferon regulatory factor (IRF), and has been demonstrated to inhibit type I/II interferon signal transduction and transform NIH3T3 cells through the interactions with IRF-1, IRF 3, and CBP/p300 proteins. To counteract vIRF's pathogenic role, we have developed five ribozymes targeting ORF-K9 mRNA to suppress vIRF expression. The vIRF RNA substrates were cleaved up to 80% in a substrate-specific manner in transcript cleavage assays in vitro. In a transient transfection assay, two of the ribozymes efficiently suppressed the expression of vIRF protein measured by dual-color immunofluorescence assay that simultaneously detects the expression of both vIRF protein and ribozyme. Flow cytometry analysis showed that these ribozymes reduced vIRF expression up to 76%. A mutant ribozyme had no cleavage activity in vitro, but exhibited antisense effect in vivo. These results suggest that the ribozymes may provide a new approach for functional knockout of vIRF gene, and are potential candidates of antiviral therapy for KSHV-related malignancies. PMID- 11393282 TI - Specific targeting of cytosine deaminase to solid tumors by engineered Clostridium acetobutylicum. AB - The presence of severe hypoxia and necrosis in solid tumors offers the potential to apply an anaerobic bacterial enzyme/prodrug approach in cancer treatment. In this context the apathogenic C. acetobutylicum was genetically engineered to express and secrete E. coli cytosine deaminase (CDase). Considerable levels of functional cytosine deaminase were detected in lysates and supernatants of recombinant C acetobutylicum cultures. After administration of the recombinant Clostridium to rhabdomyosarcoma bearing rats used as a model, cytosine deaminase could be detected at the tumor site. Moreover, following administration of the vascular targeting agent combretastatin A-4 phosphate significantly increased levels of cytosine deaminase were detected at the tumor site as a consequence of enlarged tumor necrosis and subsequently improved growth of C. acetobutylicum. The results provide evidence for the potential application of Clostrisdium-based therapeutic protein transfer to tumors in anticancer therapy. PMID- 11393283 TI - Evaluation of a new dual-specificity promoter for selective induction of apoptosis in breast cancer cells. AB - The conditional expression of lethal genes in tumor cells is a promising gene therapy approach for the treatment of cancer. The identification of promoters that are preferentially active in cancer cells is the starting point for this strategy. The combination of tissue-specific and tumor-specific elements offers the possibility to artificially develop such promoters. We describe the construction and characterization of a hybrid promoter for transcriptional targeting of breast cancer. In many cases, breast cancer cells retain the expression of estrogen receptors, and most solid tumors suffer from hypoxia as a consequence of their aberrant vascularization. Estrogen response elements and hypoxia-responsive elements were combined to activate transcription in cells that present at least one of these characteristics. When a promoter containing these elements is used to control the expression of the pro-apoptotic gene harakiri, the induction of cell death can be activated by estrogens and hypoxia, and inhibited by antiestrogens such as tamoxifen. Finally, we show evidence that these properties are maintained in the context of an adenoviral vector (AdEHhrk). Therefore, infection with this virus preferentially kills estrogen receptor positive breast cancer cells, or cells growing under hypoxic conditions. We propose the use of this promoter for transcriptional targeting of breast cancer. PMID- 11393284 TI - Herpes simplex virus thymidine kinase/ganciclovir-induced cell death is enhanced by co-expression of caspase-3 in ovarian carcinoma cells. AB - There is a need to enhance the efficacy of genetic prodrug activation therapy using herpes simplex virus thymidine kinase (tk) and ganciclovir (GCV) following disappointing results in early clinical trials. tk/GCV has been shown to lead to the activation of caspase-3, a potent executor of apoptosis. We demonstrate that co-expression of pro-caspase-3 with tk/GCV leads to enhanced cell death in ovarian carcinoma cells in vitro. Following transfection with recombinant adenoviral vectors encoding tk, GCV treatment leads to greater cell death in pro caspase-3-expressing clones of SKOV3 and IGROV1 than control cells, as well as more rapid activation of caspase-3 and more rapid cleavage of PARP. Flow cytometry suggests that there is a greater degree of S-phase block in the pro caspase-3-expressing clones than in control cells following treatment with tk/GCV. None of these effects is seen following transfection with a control adenovirus that does not encode tk. The increased cell death, early caspase-3 activation and PARP cleavage, and flow cytometric changes seen in pro-caspase-3 expressing cells can be partially inhibited by treatment with benzyloxycarbonyl val-ala-asp fluoromethylketone, a synthetic caspase inhibitor. Our data suggest that co-expression of pro-caspase-3 may lead to a significant enhancement of the efficacy of tk/GCV therapy. PMID- 11393285 TI - Helicobacter pylori: towards new therapeutic targets. PMID- 11393286 TI - Immunogenicity and protective role of an IgA reactive 31-kDa antigen of Vibrio cholerae O139. AB - Immunoglobulin A (IgA) is important in protective immunity against infection by Vibrio cholerae. In this study, the immune response to and protective role of a 31-kDa antigen of V. cholerae O139, reacting with IgA antibodies present in the sera of cholera patients and common to V. cholerae strains O139 and O1 was evaluated in BALB/c mice. From the various antigens of V. cholerae O139 and V. cholerae O1 which reacted with IgA antibodies in sera of a cholera patient, a 31 kDa common antigen was selected and purified by DEAE-Sepharose CL 6B column chromatography. Oral administration of live V. cholerae O139 in BALB/c mice elicited an IgA response to the 31-kDa antigen in serum and intestinal fluid, and a proliferation of the splenic lymphocytes on stimulation with the same antigen. The cytokine profile of these splenic lymphocytes revealed a shift from a mixed Th1 and Th2 response--interleukin-10 (IL-10) and interferon-gamma--in the first week after infection to a Th2 type of response--IL-10--in the third week. In passive protection studies, hyperimmune serum to the 31-kDa antigen was able to protect infant mice against challenge with O139 and O1 strains. These results demonstrate the ability of the 31-kDa antigen of V. cholerae O139 to induce humoral and cellular immune responses in mice, and its immunoprotective nature. PMID- 11393287 TI - Immunochemical characterisation of Vibrio cholerae O139 O antigens and production of a diagnostic antiserum without absorption. AB - Rabbits and mice immunised with chemically extracted O-antigens (O-Ags) of Vibrio cholerae O139 (O-AgB and O-AgD) developed antibodies (Abs) which appeared to be highly specific in ELISA for the relevant antigens and V. cholerae O139 strains without absorption, in contrast to the Abs against the heated O-Ag (O-AgH). An ELISA test based on the use of these Abs was shown to detect V. cholerae O139 strains down to concentrations of (9.4 x 10(4))-(7.5 x 10(5)) vibrios/ml and demonstrated no cross-reaction with other vibrios including representatives of serogroup O22. Native and proteinase K-treated O-AgB, O-AgD, O-AgH, as well as whole-cell lysates of V. cholerae O139 strains of different origin were used in immunoblotting with these Abs. Clear differences in the patterns of zones of specific reaction between chemically extracted and heated O-Ags and between lipopolysaccharide profiles of the V. cholerae O139 strains of different origin were observed. Serogroup-specific protein bands in the native O-AgB and O-AgD preparations were defined. The approach described for obtaining serogroup specific Abs against vibrios and other bacteria seems to be promising for the development of specific diagnostic tests and further investigation of bacterial antigenic structure. PMID- 11393288 TI - Detection of Legionella pneumophila antigen in urine samples by the BinaxNOW immunochromatographic assay and comparison with both Binax Legionella Urinary Enzyme Immunoassay (EIA) and Biotest Legionella Urin Antigen EIA. AB - The new BinaxNOW Immunochromatographic (ICT) Assay for the detection of Legionella pneumophila antigens was used to test 535 urine specimens from patients with and without Legionnaires' disease. The specificity, calculated by testing 112 samples from patients with pneumonia of aetiologies other than Legionella infection, and 167 urine specimens from urinary tract infections, was found to be 97.1% if the manufacturer's guidelines were followed. However, it was determined that the 'false positive' results characterised by very weak bands could be discounted by re-examination of the results at 60 min, yielding a specificity of 100%. With this minor modification of the procedure applied to examination of urine samples from 117 patients with legionellosis confirmed by isolation of L. pneumophila and 70 patients who had seroconverted to L. pneumophila serogroup 1, sensitivity was calculated to be 79.7%. In comparison, the sensitivities of the Binax Urinary Antigen Enzyme Immunoassay (EIA) and Biotest Urin Antigen EIA were estimated to be 79.1 and 83.4%, respectively. Eleven cases (5.9%) were positive by BinaxNOW assay but negative by Binax or Biotest EIA, or both. The sensitivities of all assays increased to c. 94% if only diagnosis of cases confirmed by isolation of serogroup 1 L. pneumophila was considered, although the sensitivity for infections caused by L. pneumophila serogroup 1 monoclonal antibody (MAb) subgroup Bellingham was significantly lower than for other MAb subgroups. The Biotest EIA recognised 10 (45%) of the 22 cases not caused by L. pneumophila serogroup 1, whereas the two Binax kits detected only three each. The ICT assay BinaxNOW can be recommended as a rapid specific test for the diagnosis of Legionnaires' diseases caused by L. pneumophila serogroup 1, although very weak bands should be interpreted cautiously. PMID- 11393289 TI - Inhibition of oxidative burst and chemotaxis in human phagocytes by Legionella pneumophila zinc metalloprotease. AB - Legionella pneumophila produces several extracellular proteins, but their role in the pathogenesis of Legionnaires' disease is unclear. This study examined the effects of the L. pneumophila major secretory protein (Msp), a zinc metalloprotease, on the oxidative burst and chemotaxis of human phagocytes. Polymorphonuclear leucocytes (PMNLs) and adherent monocytes treated with sublethal amounts of Msp protease were stimulated with formyl-leucyl-methionyl phenylalanine (fMLP) and opsonised zymosan particles (ZAP). A dose-dependent inhibition in superoxide anion production in response to both stimuli was seen, and complete inhibition was achieved in PMNLs and monocytes treated with Msp at concentrations of 1500 and 1000 U/ml, respectively. ZAP-induced chemiluminescence by PMNLs and mononuclear cells and fMLP-induced PMNL nitroblue tetrazolium dye reduction were both significantly inhibited. The chemotactic response of PMNLs to fMLP was inhibited in a dose-dependent manner and substantial inhibition (11% of control) was achieved with Msp 1200 U/ml. These results suggest that the L. pneumophila Msp protease alters human phagocyte functional responses significantly and may contribute to the pathogenesis of Legionnaires' disease. PMID- 11393290 TI - Flow cytometric analysis of Clostridium difficile adherence to human intestinal epithelial cells. AB - Clostridium difficile is the most common cause of diarrhoea in hospitalised patients. Bacterial adherence to gut epithelial cells is a likely prerequisite to infection and toxin production. A novel flow cytometric method was developed for detecting adherence of C. difficile to human colonic and small intestinal epithelial cells (EC) and human intestinal cell lines. Small intestinal and colonic EC were isolated from biopsy specimens with mucolytic and chelating agents. Adherence of fluorochrome-labelled C. difficile to EC was measured by flow cytometry and was calculated as increase in median fluorescent intensity (deltaMFI). Cells with bacteria attached could be distinguished easily from cells alone or cells with unlabelled bacteria attached. Toxin-positive C. difficile adhered to colonic and small intestinal EC (deltaMFI mean 21.2 SD 16.7, n = 33 and 16.5 SD 20.7, n = 19 respectively). The toxin-negative strain also adhered to both epithelial cell types (deltaMFI 26.1 SD 32.5, n = 17 and 18.3 SD 31.3, n = 16). Adherence of toxin-positive C. difficile to the intestinal cell lines Caco-2 (deltaMFI 9.4 SD 4.4, n = 14) and HT29 (deltaMFI 8.1 SD 3.1, n = 12) was quantifiable, although at a significantly lower level than with primary colonic epithelial cells. Adherence of the toxin-negative strain was slightly lower, deltaMFI 6.5 SD 1.8, n = 9 with Caco-2 cells and deltaMFI 6.0 SD 2.0, n = 10 with HT29 cells. Adherence of C. difficile to epithelial cell lines was blocked with C. difficile antiserum, confirming specificity of adherence. In conclusion, flow cytometry is a useful approach to quantifying adherence of C. difficile to human colonic and small intestinal epithelial cells. Binding of toxin-negative as well as toxin-positive bacteria was detectable by this approach. Analysis of C. difficile adherence to target cells may have important implications for the understanding of the pathogenesis of C. difficile-related disease. PMID- 11393291 TI - Immunolocalisation of Burkholderia cepacia in the lungs of cystic fibrosis patients. AB - Infection by Burkholderia cepacia is sometimes fatal in patients with cystic fibrosis (CF), as the organism can cause necrotising pneumonia and septicaemia (the cepacia syndrome), and is resistant to antibiotics. To increase knowledge of the pathogenesis of lung infection, the present study investigated the distribution of B. cepacia in lung explants from nine CF recipients of double lung transplants, of which six were colonised with both B. cepacia and Pseudomonas aeruginosa and the other three with P. aeruginosa only. In one case, explants of the donor lung (allograft) became available after the patient succumbed post-operatively to the cepacia syndrome. Further autopsy sections were examined from two patients who had chronic and then acute infection with B. cepacia. A specific antibody reactive with all five genomovars of the B. cepacia complex and another antibody specific for the 22-kDa adhesin of cable pili, were used to localise bacteria in situ. In chronic infection, the organisms were diffusely distributed, but most concentrated in hyperplastic bronchiolar epithelium, inflamed peribronchial and perivascular areas, between adjacent airway epithelial cells and in pathologically thickened alveolar septae and luminal macrophages. In acute infections the distribution was more focal, with B. cepacia on injured airway surfaces and in sites of pneumonia and abscess formation. In autopsy sections from one of the patients with chronic, then acute infection, B. cepacia was also observed in the lumen of blood capillaries. These results suggest that B. cepacia has the capacity to be highly invasive, migrating from the airways across the epithelial barrier to invade the lung parenchyma and capillaries, thereby initiating septicaemia. PMID- 11393292 TI - Isolation and characterisation of a 17-kDa staphylococcal heparin-binding protein with broad specificity. AB - A previous study reported the ability of staphylococci to bind heparin and heparin-dependent host growth factors. The present study isolated and identified heparin- and basic fibroblast growth factor (bFGF)-binding surface components of S. epidermidis strain RP12 and S. haemolyticus strain SM 131. The staphylococcal heparin-binding component(s) were purified by affinity chromatography on heparin Sepharose and a major heparin-binding protein, here designated HBP, was identified by immunoblot in these two coagulase-negative staphylococcal (CNS) species. The HBP was shown to be acidic with an approximate pI of 4.6 and a molecular mass around 17 kDa. The binding of heparin to HBP was inhibited by heparin, fucoidan, pentosan polysulphate and various other sulphated polysaccharides, but not by non-sulphated compounds. However, the purified HBP from both S. epidermidis and S. haemolyticus revealed broad specificity, and also bound bFGF, thrombospondin, von Willebrand factor and, weakly, fibrinogen. The N terminal sequences of the 17-kDa HBP from S. epidermidis and S. haemolyticus showed only limited identity. Comparison of the first 15 amino acid residues derived from either strain with known sequences in the protein databases revealed no close similarities. Taken together, these results suggest that the adhesion of at least some CNS to host sulphated glycosaminoglycans may be mediated by a previously uncharacterised group of surface proteins. PMID- 11393293 TI - Persistence of a clone of methicillin-resistant Staphylococcus aureus in a burns unit. AB - A total of 128 MRSA isolates from a burns unit in 1992 and 1997 was studied by resistotyping, plasmid analysis and pulsed-field gel electrophoresis (PFGE) of SmaI-digested chromosomal DNA to ascertain whether a clone of MRSA had persisted in the unit or whether different clones had been introduced at different times. All the MRSA isolates produced beta-lactamase and had high MICs to methicillin (>256 mg/L). All were resistant to tetracycline, kanamycin, cadmium acetate and mercuric chloride. Most were resistant to gentamicin, neomycin, erythromycin, chloramphenicol, trimethoprim, ciprofloxacin, propamidine isethionate and ethidium bromide, and were susceptible to minocycline, vancomycin and teicoplanin. None of the 1992 isolates was resistant to mupirocin, but 56% and 19% of the 1997 isolates expressed high- and low-level mupirocin resistance, respectively. Many of the 1997 isolates had acquired a 38-kb plasmid encoding high-level mupirocin resistance. The 1992 isolates had two main PFGE patterns; 82% of them belonged to PFGE pattern 1. The 1997 isolates had PFGE pattern 1, the same as the majority of the 1992 isolates. All MRSA isolates from both years carried the mecA gene in the same SmaI fragment. These findings demonstrated that a clone of MRSA that was prevalentin the burns unit in 1992 had persisted and became the predominant clone in 1997. PMID- 11393294 TI - A comparison of the bactericidal activity of quinolone antibiotics in a Mycobacterium fortuitum model. AB - New agents are urgently needed to meet the threat of multiple drug-resistant tuberculosis and to manage infection with the naturally resistant non tuberculosis mycobacteria. Earlier fluoroquinolones have been shown to have promising in-vitro activity, although mouse infection and clinical studies suggested that they lack sufficient bactericidal activity. Methods were evaluated to measure the bactericidal activity of fluoroquinolones and to compare the new agent moxifloxacin with other fluoroquinolones with M. fortuitum as a model system. The optimum bactericidal concentrations (OBC) for the fluoroquinolones were: moxifloxacin, 0.5 mg/L; ciprofloxacin and sparfloxacin, 2 mg/L and ofloxacin, 8 mg/L. The bactericidal indices (BI) for moxifloxacin, ciprofloxacin, sparfloxacin and ofloxacin were 1.8, 0.5, 0.2 and 0.2, respectively. Similar ranking was obtained when the time taken to produce one log10 reduction in viable count was calculated. These data indicate that moxifloxacin was the most bactericidal of the fluoroquinolones tested. Such methods provide a simple in vitro measure that correlates with in-vivo models. PMID- 11393295 TI - A modified rapid method of nucleic acid isolation from suspension of matured virus: applied in restriction analysis of DNA from an adenovirus prototype strain and a patient isolate. AB - This report describes a method for the isolation of nucleic acid from a suspension of matured virus. Nucleic acid (DNA) was isolated from a prototype strain of adenovirus type 7 and a clinical isolate of adenovirus type 7. Instead of the usual method of ultracentifugation, a filtration method was applied to concentrate the virus rapidly and nucleic acid was then isolated by a standard phenol/chloroform/isoamyl-alcohol extraction procedure. The DNA was found to be sufficiently purified to generate a reproducible restriction endonuclease digestion pattern. The clinical isolate of adenovirus type 7 revealed loss of restriction site for the endonuclease HindIII when compared with the prototype strain. PMID- 11393296 TI - Plasmids of mycobacteria. PMID- 11393297 TI - Competitive inhibition ELISA for the detection of Borrelia burgdorferi antigens- failure to detect antigen in the cerebrospinal fluid from patients with neuroborreliosis. PMID- 11393298 TI - Testing hypotheses on specific environmental causal effects on behavior. AB - There have been strong critiques of the notion that environmental influences can have an important effect on psychological functioning. The substance of these criticisms is considered in order to infer the methodological challenges that have to be met. Concepts of cause and of the testing of causal effects are discussed with a particular focus on the need to consider sample selection and the value (and limitations) of longitudinal data. The designs that may be used to test hypotheses on specific environmental risk mechanisms for psychopathology are discussed in relation to a range of adoption strategies, twin designs, various types of "natural experiments," migration designs, the study of secular change, and intervention designs. In each case, consideration is given to the need for samples that "pull-apart" variables that ordinarily go together, specific hypotheses on possible causal processes, and the specification and testing of key assumptions. It is concluded that environmental risk hypotheses can be (and have been) put to the test but that it is usually necessary to use a combination of research strategies. PMID- 11393299 TI - Dietary variety, energy regulation, and obesity. AB - Increased variety in the food supply may contribute to the development and maintenance of obesity. Thirty-nine studies examining dietary variety, energy intake, and body composition are reviewed. Animal and human studies show that food consumption increases when there is more variety in a meal or diet and that greater dietary variety is associated with increased body weight and fat. A hypothesized mechanism for these findings is sensory-specific satiety, a phenomenon demonstrating greater reductions in hedonic ratings or intake of foods consumed compared with foods not consumed. Nineteen studies documenting change in preference, intake, and hedonic ratings of food after a food has been eaten to satiation in animals and humans are reviewed, and the theory of sensory-specific satiety is examined. The review concludes with the relevance of oral habituation theory as a unifying construct for the effects of variety and sensory-specific satiety, clinical implications of dietary variety and sensory-specific satiety on energy regulation, and suggestions for future research. PMID- 11393300 TI - A century later: Woodworth's (1899) two-component model of goal-directed aiming. AB - In 1899, R. S. Woodworth published a seminal monograph, "The Accuracy of Voluntary Movement." As well as making a number of important empirical contributions, Woodworth presented a model of speed-accuracy relations in the control of upper limb movements. The model has come to be known as the two component model because the control of speeded limb movements was hypothesized to entail both a central and a feedback-based component. Woodworth's (1899) ideas about the control of rapid aiming movements are evaluated in the context of current empirical and theoretical contributions. PMID- 11393301 TI - Brain-computer communication: unlocking the locked in. AB - With the increasing efficiency of life-support systems and better intensive care, more patients survive severe injuries of the brain and spinal cord. Many of these patients experience locked-in syndrome: The active mind is locked in a paralyzed body. Consequently, communication is extremely restricted or impossible. A muscle independent communication channel overcomes this problem and is realized through a brain-computer interface, a direct connection between brain and computer. The number of technically elaborated brain-computer interfaces is in contrast with the number of systems used in the daily life of locked-in patients. It is hypothesized that a profound knowledge and consideration of psychological principles are necessary to make brain-computer interfaces feasible for locked-in patients. PMID- 11393302 TI - The job satisfaction-job performance relationship: a qualitative and quantitative review. AB - A qualitative and quantitative review of the relationship between job satisfaction and job performance is provided. The qualitative review is organized around 7 models that characterize past research on the relationship between job satisfaction and job performance. Although some models have received more support than have others, research has not provided conclusive confirmation or disconfirmation of any model, partly because of a lack of assimilation and integration in the literature. Research devoted to testing these models waned following 2 meta-analyses of the job satisfaction-job performance relationship. Because of limitations in these prior analyses and the misinterpretation of their findings, a new meta-analysis was conducted on 312 samples with a combined N of 54,417. The mean true correlation between overall job satisfaction and job performance was estimated to be .30. In light of these results and the qualitative review, an agenda for future research on the satisfaction-performance relationship is provided. PMID- 11393303 TI - The error of accepting the "theoretical" null hypothesis: the rise, fall, and resurrection of commonsense hypotheses in psychology. AB - When psychologists test a commonsense (CS) hypothesis and obtain no support, they tend to erroneously conclude that the CS belief is wrong. In many such cases it appears, after many years, that the CS hypothesis was valid after all. It is argued that this error of accepting the "theoretical" null hypothesis reflects confusion between the operationalized hypothesis and the theory or generalization that it is designed to test. That is, on the basis of reliable null data one can accept the operationalized null hypothesis (e.g., "A measure of attitude x is not correlated with a measure of behavior y"). In contrast, one cannot generalize from the findings and accept the abstract or theoretical null (e.g., "We know that attitudes do not predict behavior"). The practice of accepting the theoretical null hypothesis hampers research and reduces the trust of the public in psychological research. PMID- 11393304 TI - Does psi exist? Comments on Milton and Wiseman's (1999) meta-analysis of ganzfeld research. AB - J. Milton and R. Wiseman (1999) attempted to replicate D. Bem and C. Honorton's (1994) meta-analysis, which yielded evidence that the ganzfeld is a suitable method for demonstrating anomalous communication. Using a database of 30 ganzfeld and autoganzfeld studies, Milton and Wiseman's meta-analysis yielded an effect size (ES) of only 0.013 (Stouffer Z = 0.70, p = .24, one-tailed). Thus they failed to replicate Bem and Honorton's finding (ES = 0.162, Stouffer Z = 2.52, p = 5.90 x 10(-3), one-tailed). The authors conducted stepwise performance comparisons between all available databases of ganzfeld research, which were argued not to be lacking in quality. Larger aggregates of such studies were formed, including a database comprising 79 ganzfeld-autoganzfeld studies (ES = 0.138, Stouffer Z = 5.66, p = 7.78 x 10(-9)). Thus Bem and Honorton's positive conclusion was confirmed. More accurate population parameters for the ganzfeld and autoganzfeld domains were calculated. Significant bidirectional psi effects were also found in all databases. The ganzfeld appears to be a replicable technique for producing psi effects in the laboratory. PMID- 11393305 TI - Copper, iron, and zinc contents in human milk during the first three months of lactation: a longitudinal study. AB - The aim of the study has been to analyze the evolution of copper, iron, and zinc contents in human milk, from colostrum to the third postpartum month, following a longitudinal design, under specific conditions of sample collection and to apply an analytical procedure previously optimized to reduce any variation outside physiological lactation. The copper, iron, and zinc concentrations in 144 milk samples from 39 healthy puerpera women, were analyzed in five stages by flame atomic absorption spectrometry, following a standardized protocol. Copper presented a gradual decrease from 0.38 mg/L to 0.19 mg/L by the 90th day; the particular analysis from colostrum to transitional milk manifested the following two tendencies. Whereas an increase from 0.19 to 0.42 mg/L was observed in some women, a decrease from 0.53 to 0.45 mg/L was detected in others; therefore, copper presented two significant behaviors in the evolution from colostrum to transitional milk. In both cases, the evaluated changes were significant. The iron content varied from 0.56 to 0.40 mg/L by the 30th day, remaining constant until the first trimester concluded. The average zinc concentration decreased sharply from 7.99 to 3.3 mg/L on d 15; the rate of decrease slowed down gradually until 1.05 mg/L. PMID- 11393306 TI - Increases of calcium and magnesium and decreases of phosphorus and iron with aging in human uterine tubes. AB - To elucidate compositional changes of the uterine tube by aging, the authors studied age-related changes of elements in human uterine tubes by inductively coupled plasma-atomic emission spectrometry. The uterine tubes were resected postmortem or surgically removed from patients with uterine myoma. It was found that the contents of calcium and magnesium increased progressively with aging in uterine tubes, whereas the contents of phosphorus and iron decreased gradually with aging. The sulfur content of uterine tubes remained constant and independent of aging. Regarding relationships between elements, significant relationships were found between calcium and magnesium contents, between phosphorus and iron contents, between phosphorus and sulfur contents, and between phosphorus and sodium contents in human uterine tubes. PMID- 11393307 TI - Host selenium status selectively influences susceptibility to experimental viral myocarditis. AB - The purpose of the present work was to determine whether dietary selenium (Se) deficiency could influence the injurious effect of human viruses other than Coxsackie virus B3 (CVB3) on mouse heart. Weanling C3H/HeN mice were fed a Se deficient or Se-adequate diet for 4 wk and then were inoculated intraperitoneally with one of the following viruses: Coxsackie virus B1 (CVB1), echovirus 9 (EV9), Coxsackie virus A9 (CVA9), or herpes simplex 1 (HSV1). Polio virus 1 (PV1) was employed as a negative control. Prior to inoculation, mean serum Se levels were 430 versus 61 ng/mL in adequate versus deficient mice, respectively. Ten days later, hearts were removed and processed by routine histological procedures. Cardiac lesions were scored according to the number and size of myocarditic foci. Significantly greater heart damage resulting from CVB1 and EV9 was observed in Se deficient than in Se-adequate mice, and the Se status had no influence on CVA9 induced myocarditis. In contrast, heart damage caused by HSV1 was significantly milder in Se-deficient than in Se-adequate mice. Therefore, it may be concluded that the Se status of the murine host selectively influences the degree of viral induced myocarditic lesions. PMID- 11393308 TI - Serum magnesium, copper, and zinc alterations following spinal fusion. AB - Changes in serum magnesium, copper and zinc values were evaluated in spinal fusion patients at four monitorings. For magnesium and copper individually, a significant difference was found between the mean values at each monitoring (p < 0.05), whereas the changes of zinc values between four monitorings were insignificant. There was no statistical difference between the changes of each trace element values and gender, age, operative time, intraoperative blood loss, blood replacement, number of the vertebral levels fused, and antibiotic type used. As a result, magnesium can be suggested to be more important than the other two elements in the postoperative period. Alterations of serum magnesium, copper, and zinc values do not have any correlation with the fusion of the spinal column, either with the width of the fused area or operative time and blood loss. This study cannot confirm the exact reason for this entity and the etiology remains speculative. There is no need for magnesium, copper, or zinc supplementation during the surgical period for the patients. It will be worthy to evaluate the patients who were sent to the intensive care unit after spinal surgery and compare their results with the other intensive care patients. PMID- 11393309 TI - Serum manganese concentrations in a representative sample of the Canarian population. AB - Serum manganese (Mn) concentrations of 368 individuals 6-75 yr of age (179 males and 189 females) living in the Canary Islands were determined by graphite furnace atomic absorption spectrometry. The mean manganese concentration was 1.06+/-0.62 microg/L, ranging between 0.19 and 3.33 microg/L. Most of the analyzed samples (63.3% of the samples) fall within the reference interval (0.54-1.76 microg/L) for apparently healthy people. Individuals from Fuerteventura presented with mean Mn concentrations significantly higher than individuals from the rest of the islands. This could be attributed to differences in the Mn content of soil and/or differences in dietary habits of the population. Serum Mn concentration did not vary with gender, and individuals younger than 18 yr old had the highest mean Mn concentration, compared to the rest of the age groups considered. No relation to socioeconomic status, educational level, and tobacco or alcohol consumption was found. However, the serum Mn concentration tended to decrease when increasing the consumption of wine or beer. Sportsmen presented significantly higher serum Mn concentrations than the rest. PMID- 11393310 TI - The effect of aging on the mineral status of female SAMP1 and SAMR1. AB - The effect of aging on the status of macrominerals and trace elements in tissues was studied using two strains (SAMP1 and SAMR1) of senescence accelerated mouse. Two-month-old, 6-mo-old, and 10-mo-old female SAMP1 and SAMR1 mice were fed a commercial diet. Iron, zinc, copper, calcium, magnesium, phosphorus, sulfur, sodium, and potassium concentrations in blood, liver, kidney, brain, and tibia of the mice were determined. The copper concentration in the brain was significantly increased with age in SAMP1 and SAMR1. In addition, the brain copper levels in SAMP1 were significantly higher than that in SAMR1 at respective ages. The calcium concentration in the kidney was significantly increased with age, but the copper and phosphorus concentrations significantly decreased with age in SAMP1 and SAMR1. In the liver of SAMR1, all minerals measured in this study except for sodium and potassium were significantly decreased with age. In addition, all mineral concentrations in the liver of 2-mo-old mice in SAMR1 except for copper and sodium were markedly higher than those in SAMP1 of the same age. These results suggest that the genetic factor is related to the age-associated mineral changes in tissues. PMID- 11393311 TI - Changes in Ca, Cu, Fe, Mg, and Zn contents in mouse brain tissues after prolonged oral ingestion of brick tea liquor containing a high level of Al. AB - A study was conducted to analyze the regional distribution of Ca, Cu, Fe, Mg, and Zn contents in brain tissues after animals were given liquor of brick tea that contained a high Al content. In 25 normal adult male mice given either water or 0.9% NaCl for 1 mo or 2 mo, the metal concentrations in the serum, liver, frontal cortex, hippocampus, cerebellum, and brain stem were comparable (p > 0.05). When the drinking water was replaced by a 1% brick tea liquor, which contained a high Al content, serum Al concentration was increased significantly 1 mo after the onset of the experiment and remained high at the end of the second month. The level of Al was also elevated in both the cortex and hippocampus at 1 mo after replacing tea for drinking water. In addition to Al, there were a significant increase in hippocampal Zn and a decrease in Cu contents. There was no change in tissue Mg or Fe contents, but there was a significant increase in Ca content in every brain region studied. It was suggested that the increase in Ca might be the result of the effect of other components in tea. Unlike the brain, there was no change in the concentration of any of the metals, including Al, in the liver, which further demonstrated that the changes observed in the brain was specific. The results of the present study confirmed that Al, when given orally in the form of tea, could be absorbed into the bloodstream. The absorbed Al could accumulate in selected brain regions. The presence of Al might also change the tissue content of endogenous trace metals. PMID- 11393312 TI - Simultaneous measurement of the trace elements Al, As, B, Be, Cd, Co, Cu, Fe, Li, Mn, Mo, Ni, Rb, Se, Sr, and Zn in human serum and their reference ranges by ICP MS. AB - The goal of this article was to establish reference ranges of the concentration of trace elements in human serum and to compare these results with those reported by other authors. We describe the sample preparation and measurement conditions that allow the rapid, precise, and accurate determination of Al, As, B, Be, Cd, Co, Cu, Fe, Li, Mn, Mo, Ni, Rb, Se, Sr, and Zn in human serum samples (n = 110) by inductively coupled plasma-mass spectrometry (ICP-MS). Accuracy and precision were determined by analyzing three reconstituted reference serum samples by comparison with other methods and by the standard addition procedure. The advantages of the ICP-MS method include short time of analysis of the elements mentioned, low detection limit, high precision, and high accuracy. Disadventages include a high risk of contamination due to the presence of some of the elements of interest in the environment, the relatively delicate sample handling, and the high cost of the equipment. PMID- 11393313 TI - A paradigm of international environmental law: the case for controlling the transboundary movements of hazardous wastes. AB - The production of large quantities of wastes globally has created a commercial activity involving the transfrontier shipments of hazardous wastes, intended to be managed at economically attractive waste-handling facilities located elsewhere. In fact, huge quantities of hazardous wastes apparently travel the world in search of "acceptable" waste management facilities. For instance, within the industrialized countries alone, millions of tonnes of potentially hazardous waste cross national frontiers each year on their way for recycling or to treatment, storage, and disposal facilities (TSDFs) because there is no local disposal capacity for these wastes, or because legal disposal or reuse in a foreign country may be more environmentally sound, or managing the wastes in the foreign country may be less expensive than at home. The cross-boundary traffic in hazardous wastes has lately been under close public scrutiny, however, resulting in the accession of several international agreements and laws to regulate such activities. This paper discusses and analyzes the most significant control measures and major agreements in this new commercial activity involving hazardous wastes. In particular, the discussion recognizes the difficulties with trying to implement the relevant international agreements among countries of vastly different socioeconomic backgrounds. Nonetheless, it is also noted that global environmental agreements will generally be a necessary component of ensuring adequate environmental protection for the world community-and thus a need for the careful implementation of such agreements and regulations. PMID- 11393314 TI - An ecological perspective on in-stream temperature: natural heat dynamics and mechanisms of human-caused thermal degradation. AB - While external factors (drivers) determine the net heat energy and water delivered to a stream, the internal structure of a stream determines how heat and water will be distributed within and exchanged among a stream's components (channel, alluvial aquifer, and riparian zone/floodplain). Therefore, the interaction between external drivers of stream temperature and the internal structure of integrated stream systems ultimately determines channel water temperature. This paper presents a synoptic, ecologically based discussion of the external drivers of stream temperature, the internal structures and processes that insulate and buffer stream temperatures, and the mechanisms of human influence on stream temperature. It provides a holistic perspective on the diversity of natural dynamics and human activities that influence stream temperature, including discussions of the role of the hyporheic zone. Key management implications include: (1) Protecting or reestablishing in-stream flow is critical for restoring desirable thermal regimes in streams. (2) Modified riparian vegetation, groundwater dynamics, and channel morphology are all important pathways of human influence on channel-water temperature and each pathway should be addressed in management plans. (3) Stream temperature research and monitoring programs will be jeopardized by an inaccurate or incomplete conceptual understanding of complex temporal and spatial stream temperature response patterns to anthropogenic influences. (4) Analyses of land-use history and the historical vs contemporary structure of the stream channel, riparian zone, and alluvial aquifer are important prerequisites for applying mechanistic temperature models to develop management prescriptions to meet in-channel temperature goals. PMID- 11393315 TI - Undamming rivers: a review of the ecological impacts of dam removal. AB - Dam removal continues to garner attention as a potential river restoration tool. The increasing possibility of dam removal through the FERC relicensing process, as well as through federal and state agency actions, makes a critical examination of the ecological benefits and costs essential. This paper reviews the possible ecological impacts of dam removal using various case studies. Restoration of an unregulated flow regime has resulted in increased biotic diversity through the enhancement of preferred spawning grounds or other habitat. By returning riverine conditions and sediment transport to formerly impounded areas, riffle/pool sequences, gravel, and cobble have reappeared, along with increases in biotic diversity. Fish passage has been another benefit of dam removal. However, the disappearance of the reservoir may also affect certain publicly desirable fisheries. Short-term ecological impacts of dam removal include an increased sediment load that may cause suffocation and abrasion to various biota and habitats. However, several recorded dam removals have suggested that the increased sediment load caused by removal should be a short-term effect. Pre removal studies for contaminated sediment may be effective at controlling toxic release problems. Although monitoring and dam removal studies are limited, a continued examination of the possible ecological impacts is important for quantifying the resistance and resilience of aquatic ecosystems. Dam removal, although controversial, is an important alternative for river restoration. PMID- 11393316 TI - Land degradation: a challenge to Ethiopia. AB - Land degradation is a great threat for the future and it requires great effort and resources to ameliorate. The major causes of land degradation in Ethiopia are the rapid population increase, severe soil loss, deforestation, low vegetative cover and unbalanced crop and livestock production. Inappropriate land-use systems and land-tenure policies enhance desertification and loss of agrobiodiversity. Utilization of dung and crop residues for fuel and other uses disturbs the sustainability of land resources. The supply of inputs such as fertilizer, farm machinery and credits are very low. The balance between crop, livestock, and forest production is disturbed, and the farmer is forced to put more land into crop production. For environmentally and socially sustainable development, there is an urgent need to promote awareness and understanding of the interdependence of natural, socioeconomic, and political systems at local and national levels. Understanding the current status and causes of land degradation is very important. This paper reveals the important elements of land degradation in Ethiopia and suggests possible solutions that may help to ameliorate the situation. PMID- 11393317 TI - Evaluating nonpoint pollution policy using a tightly coupled spatial decision support system. AB - Policy makers often must rely on the cumulative impact of independent actions taken by local landowners to achieve environmental goals. The connection between policy, regulation, and local action, however, is often not well understood and, thus, the impact of proposed policies may be difficult to predict. In this study we evaluate the effectiveness of alternative policy scenarios for agricultural set aside programs (e.g., the conservation reserve program administered by the United States Department of Agriculture) in reducing nonpoint pollution. Two alternative policy scenarios are developed and analyzed; one based on the erodibility index (detachment), the other sediment yield (transport). An estimate of the cumulative impact of associated land use change on nonpoint pollution is made using the AGNPS distributed parameter watershed model. This work is completed within the Cypress Creek watershed in southern Illinois. An analysis of the resulting data suggests that the most efficacious regulatory strategy for achieving nonpoint water pollution goals depends, in part, on place-specific land use patterns. This conclusion provides a solid argument for place-based regulatory strategies. PMID- 11393318 TI - Environmental management of sulfur trioxide emission: impact of SO3 on human health. AB - The major contributors to global acidification are sulfur oxides and nitrogen oxides emitted mostly by the burning of fossil fuels. From the scientific point of view, it is necessary to make a clear distinction between sulfur dioxide and sulfur trioxide when referring to sulfur oxides. These two air pollutants have different properties. This paper reports the following aspects: the strong effect of sulfur trioxide on local human health (a case study of asthma in Yokkaichi), the problem of corrosion caused by sulfur trioxide, the difference in analytical methods for determining sulfur dioxide concentrations and sulfur trioxide concentrations, and the difference in removal methods for sulfur dioxide and sulfur trioxide. An important initiative at the third European conference of environment ministers was that the issue of human health related to local air pollution should be given priority over that of global pollution. The declines in the emissions of sulfur dioxide and nitrogen oxides have mainly been effective in reducing acidification due to long-range transport. The reduction in sulfur trioxide may be more effective in improving local human health mentioned in the initiative. PMID- 11393319 TI - Integration of visual quality considerations in development of Israeli vegetation management policy. AB - This article deals with the visual quality of Mediterranean vegetation groups in northern Israel, the public's preference of these groups as a visual resource, and the policy options for their management. The study is based on a sample of 44 Mediterranean vegetation groups and three population groups of local residents, who were interviewed using a questionnaire and photographs of the vegetation groups. The results of the research showed that plant classification methods based on flora composition, habitat, and external appearance were found to be suitable for visual plant classification and for the evaluation of visual preference of vegetation groups by the interviewed public. The vegetation groups of planted pine forests and olive groves, characterizing a cultured vegetation landscape, were preferred over typical Mediterranean landscapes such as scrub and grassed scrub. The researchers noted a marked difference between the two products of vegetation management policy, one that proposes the conservation and restoration of the variety of native Mediterranean vegetation landscape, and a second that advanced the development of the cultured landscape of planted olive groves and pines forests, which were highly preferred by the public. The authors suggested the development of an integrated vegetation management policy that would combine both needs and thus reduce the gap between the policy proposed by planners and the local population's visual preference. PMID- 11393320 TI - Identifying determinants of nations' wetland management programs using structural equation modeling: an exploratory analysis. AB - Integrated management and policy models suggest that solutions to environmental issues may be linked to the socioeconomic and political characteristics of a nation. In this study, we empirically explore these suggestions by applying them to the wetland management activities of nations. Structural equation modeling was used to evaluate a model of national wetland management effort and one of national wetland protection. Using five predictor variables of social capital, economic capital, environmental and political characteristics, and land-use pressure, the multivariate models were able to explain 60% of the variation in nations' wetland protection efforts based on data from 90 nations, as defined by level of participation in the international wetland convention. Social capital had the largest direct effect on wetland protection efforts, suggesting that increased social development may eventually lead to better wetland protection. In contrast, increasing economic development had a negative linear relationship with wetland protection efforts, suggesting the need for explicit wetland protection programs as nations continue to focus on economic development. Government, environmental characteristics, and land-use pressure also had a positive direct effect on wetland protection, and mediated the effect of social capital on wetland protection. Explicit wetland protection policies, combined with a focus on social development, would lead to better wetland protection at the national level. PMID- 11393321 TI - Home range size and choice of management strategy for lynx in Scandinavia. AB - Annual and seasonal home ranges were calculated for 47 Eurasian lynx in four Scandinavian study sites (two in Sweden and two in Norway). The observed home ranges were the largest reported for the species, with study site averages ranging from 600 to 1,400 km2 for resident males and from 300 to 800 km2 for resident females. When home range sizes were compared to the size of protected areas (national parks and nature reserves) in Scandinavia, it was concluded that very few protected areas contained sufficient forest to provide space for more than a few individuals. As a direct consequence of this, most lynx need to be conserved in the multiuse semi-natural forest habitats that cover large areas in Scandinavia. This conservation strategy leads to a number of conflicts with some land uses (sheep and semidomestic reindeer herding, and roe deer hunters), but not all (forestry and moose harvest). Accordingly research must be aimed at understanding the ecology of these conflicts, and finding solutions. PMID- 11393322 TI - Scoring corporate environmental reports for comprehensiveness: a comparison of three systems. AB - Corporate annual environmental reports can be judged by the comprehensiveness of their coverage; this paper uses three published comprehensiveness scoring systems to judge 28 such reports (mostly dated 1998) from large US corporations. Earlier (nominally 1996) reports from the same companies were previously scored using one or two of the same systems, and the published results are compared with scores from the current round of reports. The scores of all three scoring systems are significantly correlated for the 1998 environmental reports, indicating that to some degree they are scoring the same features, but with quite a lot of scatter between the scores of one of the systems and those of the other two. There is, however, no correlation between the scores on the 1996 reports and the 1998 reports using the same systems on both. Evidently, most of the 1998 reports were not prepared with the intent of maximizing scores from any of the three scoring systems. Although the three systems achieve similar ranking of the 1998 reports for environmental comprehensiveness, the average normalized scores of one of the systems is significantly higher than those of the other two, reflecting a shorter list of topics and one more in keeping with the practices of the report writers. Because the scoring systems measure the number of topics covered and the depth of discussion rather than the quality of environmental performance, maximum scores could be obtained even with poor performance. PMID- 11393323 TI - An investigation of environmental racism claims: testing environmental management approaches with a geographic information system. AB - The purpose of this research was to explore the concept of an environmental racism claim through the use of several environmental management tools. The EPAs Toxics Release Inventory, Cumulative Exposure Project, and the Los Angeles County Department of Health Services' Hot Zone Census Tract Assessment were combined with racial and socioeconomic data to test claims that minorities in South Central Los Angeles are disproportionately exposed to environmental lead. Multivariate analysis indicated that race is strongly associated with the number of cases of elevated blood lead levels in South Central, irrespective of poverty status. Proximity to point sources, a common focal point for studies of environmental racism, was not a contributing factor to health outcomes. Proximity to transportation corridors was consistently the strongest indicator of environmental lead exposure, while median home values were significantly and positively related to elevated blood lead levels. Implications for environmental justice advocates and social and environmental scientists are discussed. PMID- 11393324 TI - Economic and environmental analysis of retrofitting a large office building with energy-efficient lighting systems. AB - An evaluation of the economic and environmental costs and benefits that would result if the Zorinsky Federal Building, located in Omaha, Nebraska, USA, converted its current lighting system to a more energy-efficient system (i.e., joined the EPA's Green Lights Program) was conducted. Lighting accounts for 20-25 percent of all electricity sold in the United States. Costs considered in the study included the cost of retrofitting the building's existing lighting system and the cost of disposal of the current lamps and ballast fixtures. Benefits included a reduction of electric utility costs and a reduction of emissions of SO2, NOx, CO2, and CO from electric utility power plants. Environmental and health issues for air pollutant emissions were also addressed. The results showed that significant reductions in utility bills as well as reductions in air emissions would result from a major building converting to a more energy efficient lighting system. The results showed that conversion of this large building would reduce SO2 emissions by 14.6 tons/yr and NOx emissions by 6.3 tons/yr. In addition, the conversion would reduce annual energy costs by approximately $114,000. PMID- 11393325 TI - Role of rural school nurses in asthma management. AB - Environmental, socioeconomic, psychological and familial factors in rural communities predispose children to asthma. This is not only the case in the US but also in the UK, New Zealand and other Western countries. Asthma prevalence ranges from 2.2 to 15%. Because children spend at least 6 hours of their day in school, school health personnel must be attentive to, and skilled in managing the needs and issues faced by children with asthma while at school. Rural school nurses or their deputies need to advise children with asthma about avoiding aeroallergens from hay, smoke, dust, grain in silos and animal dander from cattle and sheep. In the case of children with asthma in rural areas, symptoms may be accepted as long as the child can go to school and play. Parents in rural areas may not believe in routine preventive care for asthma as part of public health practice. Rural nurses need to be aware of current asthma guidelines and apply the concepts to prevention. They need to be proactive and engage in primary, secondary and tertiary prevention. Rural school nurses can begin by using existing resources and adapting these resources for use in rural school environments. Worldwide asthma education is fundamental to asthma patient management. PMID- 11393326 TI - Multiple sclerosis in childhood and adolescence: clinical features and management. AB - The presentation of multiple sclerosis (MS) in childhood has traditionally been thought to be rare. However, more paediatric cases are now being reported, as a result of progress in diagnostic techniques with the use of sensitive imaging modalities of the brain and spinal cord. Management from an early age and the availability of new treatment options have changed the outcome of paediatric MS. Drugs currently available for treatment, such as beta-interferons, copolymer-1 and intravenous immunoglobulin G, have been found to reduce relapse rate, disease severity and progression to disability in adults, but have not been investigated in children and adolescents. The overall outcome of MS in children is apparently no worse than in adults and the disease may even be less aggressive in children. In juvenile MS, disease progression does not appear to be related to age of onset, severity of neurological involvement or mono/polysymptomatic involvement at presentation. The potential to treat MS has significantly changed the prognosis. Early diagnosis is important, as early treatment can prevent or delay the development of disability. PMID- 11393327 TI - Pharmacological management of pain and anxiety during emergency procedures in children. AB - Painful procedures are frequently required during treatment of children in the emergency department and are very stressful for the children, their parents and healthcare providers. Pharmacological methods to safely provide almost painless local anaesthesia, analgesia and anxiolysis have been increasingly studied in children. With knowledge of these methods, and patience, the emergency care provider can greatly reduce the distress often associated with emergency care of children. Topical local anaesthetics such as LET [lidocaine (lignocaine), epinephrine (adrenaline), tetracaine] or buffered lidocaine injected through the wound with fine needles can almost painlessly anaesthetise lacerations for suturing. Topical creams such as lidocaine/ prilocaine (EMLA) or tetracaine, iontophoresed lidocaine, or buffered lidocaine subcutaneously injected with fine needles can make intravenous catheter placement virtually 'painless'. When anxiety is significant, and mild to moderate analgesia/ anxiolysis/amnesia is needed, nitrous oxide can be administered if the proper delivery devices are available. Alternatively, when intensely painful fracture reduction, burn debridement, or abscess drainage is necessary, well tolerated and effective deep sedation can be achieved with careful use of midazolam and either ketamine or fentanyl. PMID- 11393328 TI - Tourette syndrome: clinical characteristics and current management strategies. AB - Tourette Syndrome (TS) is a disorder comprised of involuntary motor and phonic tics often associated with psychiatric conditions. The etiology for TS is unclear, with both genetic and immunological theories being studied to date. When pharmacotherapy is considered by the patient and physician to be required, owing to either functional impairment from tics or comorbid psychiatric illness, dopamine receptor antagonists are commonly used. Our first-line agents for tic suppression include clonidine, guanfacine, clonazepam and baclofen. Should these agents be ineffective, we would recommend pimozide, fluphenazine, risperidone or haloperidol. The potential benefit of other agents, such as olanzapine, ziprasidone, pergolide and botulinum toxin, is encouraging. Despite many years of concern, we have found little exacerbation of tics with stimulant medications for attention deficit hyperactivity disorder, but clearly clonidine and guanfacine can ameliorate both comorbid conditions. Obsessive compulsive disorder, when associated with TS, may be treated with either a selective serotonin reuptake inhibitor in association with a dopamine receptor antagonist or risperidone alone. New therapies for all aspects of TS and its comorbid conditions are in active clinical trials. PMID- 11393329 TI - Fluoroquinolones in paediatrics. AB - The fluoroquinolones are an important group of antibiotics, which are widely used in adult patients because of their high penetration in tissues and bactericidal activity. However, they are not licensed for paediatric use (except the limited indication of Pseudomonas infection in cystic fibrosis) because of their potential to cause joint toxicity (observed in experiments using juvenile animal models). In recent years, there has been a change in the susceptibility of pathogens to widely used antibiotics; however, many of these pathogens remain sensitive to the fluoroquinolones (agents which can often be administered orally to treat severe infections). Fluoroquinolones have a number of potential indications in children: cystic fibrosis, intestinal infections due to resistant strains of Salmonella spp. and Shigella spp., severe infections due to Enterobacteriaceae (including the neonatal period), complicated urinary tract infections, the immunocompromised host, and some mycobacterial infections. The third generation fluoroquinolones have improved activity against Gram-positive bacteria and could be useful in respiratory tract, and ear, nose and throat infections in adult patients. Their potential role in routine use for paediatric patients will remain limited because of potential joint complications and the availability of other treatment options. However, available clinical data does indicate that the incidence of arthrotoxicity in children treated with ciprofloxacin appears to be the same as that in adult patients. The use of other fluoroquinolones is too rare to obtain meaningful information on their toxicity in children. For future fluoroquinolones, pneumococcal meningitis will probably be a potential indication. Despite their important activity, fluoroquinolones remain a second-line treatment in children, for use following the failure of a well established antibiotic treatment, to avoid potential adverse effects and the emergence of resistant strains. PMID- 11393331 TI - The evolution of menopause and human life span. AB - Noteworthy data is emerging to support the existence of longevity-enabling genes. Our observations of the relationship between reproductive fitness and longevity among centenarians support theories that posit strong selective forces in the determination of how fast humans age and their susceptibility to diseases associated with ageing. Current data support the idea that there is no selective advantage for humans to have a lifespan of approximately 100 years. Rather, getting to such a very old age may be a by-product of longevity-enabling genes that maximize the length of time during which women can bear children, and during which they can increase the survival probabilities of their children and grandchildren. We thus review the literature pertaining to the relationship between reproductive fitness and longevity. PMID- 11393332 TI - Secular trend in birthweight among the Purari delta population, Papua New Guinea. AB - The aim of this analysis was to examine the extent and possible seasonal nature of the secular trend in mean birthweight in the Purari delta, Papua New Guinea. This is a country undergoing rapid modernization, and with this has come a secular trend toward increased adult body size in some parts of the country but not others. Birthweight data, collected by month of delivery at Kapuna Hospital in the Purari delta between the years 1969 and 1996, was analysed by year of birth and by season, using one-way analysis of variance and post hoc Scheffe tests with the statistical software SPSS-PC+. A total of 927 birthweights for the years 1969, 1972, 1977, 1994 and 1996 were included in the analysis. Twin births were excluded from analysis, as were births below 1.5 kg. There is clear evidence of a secular trend in increasing mean birthweight between the period 1969 and 1996, with the largest significant difference being between 1977 and 1994, from 2.70 to 2.92 kg. There were no significant differences in mean birthweight between the sexes. The rate of birthweight increase between 1977 and 1994 was 130 g per decade, lower than the gain of 200 g per decade in the period 1994-1996. The decline in birthweight of 90 g per decade during the period 1969-1977 is not statistically significant. The proportion of infants born with low birth weight (< 2.5 kg) shows an increase between 1969 and 1972, and a decline thereafter. While seasonal differences in birthweight during any of the years examined is non significant, significantly greater mean birthweight across the period 1969-1996 was found for births during the wet season (April to July), and the drier season (August to November), respectively. The secular increase in mean birthweight is likely to be a consequence of the improvements in maternal diet and increased maternal body size that have come with economic modernization. The secular trend of seasonality in mean birthweight among the Purari delta population may be a function of seasonally varied displacement of traditional diet by non-local bought foods, as well as reduced seasonality of maternal workload associated with the processing of the traditional staple food. PMID- 11393330 TI - Treatment of typical absence seizures and related epileptic syndromes. AB - Typical absences are brief (seconds) generalised seizures of sudden onset and termination. They have 2 essential components: clinically, the impairment of consciousness (absence) and, generalised 3 to 4Hz spike/polyspike and slow wave discharges on electroencephalogram (EEG). They differ fundamentally from other seizures and are pharmacologically unique. Their clinical and EEG manifestations are syndrome-related. Impairment of consciousness may be severe, moderate, mild or inconspicuous. This is often associated with motor manifestations, automatisms and autonomic disturbances. Clonic, tonic and atonic components alone or in combination are motor symptoms; myoclonia, mainly of facial muscles, is the most common. The ictal EEG discharge may be consistently brief (2 to 5 seconds) or long (15 to 30 seconds), continuous or fragmented, with single or multiple spikes associated with the slow wave. The intradischarge frequency may be constant or may vary (2.5 to 5Hz). Typical absences are easily precipitated by hyperventilation in about 90% of untreated patients. They are usually spontaneous, but can be triggered by photic, pattern, video games stimuli, and mental or emotional factors. Typical absences usually start in childhood or adolescence. They occur in around 10 to 15% of adults with epilepsies, often combined with other generalised seizures. They may remit with age or be lifelong. Syndromic diagnosis is important for treatment strategies and prognosis. Absences may be severe and the only seizure type, as in childhood absence epilepsy. They may predominate in other syndromes or be mild and nonpredominant in syndromes such as juvenile myoclonic epilepsy where myoclonic jerks and generalised tonic clonic seizures are the main concern. Typical absence status epilepticus occurs in about 30% of patients and is more common in certain syndromes, e.g. idiopathic generalised epilepsy with perioral myoclonia or phantom absences. Typical absence seizures are often easy to diagnose and treat. Valproic acid, ethosuximide and lamotrigine, alone or in combination, are first-line therapy. Valproic acid controls absences in 75% of patients and also GTCS (70%) and myoclonic jerks (75%); however, it may be undesirable for some women. Similarly, lamotrigine may control absences and GTCS in possibly 50 to 60% of patients, but may worsen myoclonic jerks; skin rashes are common. Ethosuximide controls 70% of absences, but it is unsuitable as monotherapy if other generalised seizures coexist. A combination of any of these 3 drugs may be needed for resistant cases. Low dosages of lamotrigine added to valproic acid may have a dramatic beneficial effect. Clonazepam, particularly in absences with myoclonic components, and acetazolamide may be useful adjunctive drugs. PMID- 11393333 TI - Growth of permanent mandibular teeth of British children aged 4 to 9 years. AB - PRIMARY OBJECTIVE: The aim of this study was to investigate ethnic differences and describe tooth formation of mandibular permanent teeth in a group of London children. RESEARCH DESIGN: The design was cross-sectional retrospective study. SAMPLE AND METHOD: The sample was a non-random group of healthy British children (n = 521) attending a dental hospital. The children aged between 4 and 9 years were of Bangladeshi or white Caucasian origin. Developing permanent mandibular teeth were staged from radiographs according to criteria described by Demirjian, Goldstein and Tanner (1973, Human Biology, 45, 211-227). Data were grouped in 6 month intervals and analysed using probit analysis. Formation was also expressed relative to stages of the first permanent molar (M1) and the distribution of stages tested between the groups and sexes using Mann Whitney U-test. RESULTS: Tooth formation was not significantly different between the two ethnic groups. Girls attained almost all stages of tooth formation earlier than boys; in addition, the canine showed significant advancement relative to M1 formation in girls (p < 0.05). CONCLUSIONS: These findings failed to demonstrate an ethnic difference in tooth formation in these children. PMID- 11393334 TI - Spatial patterns of child growth in Papua New Guinea and their relation to environment, diet, socio-economic status and subsistence activities. AB - Anthropometric data from the 1982/83 Papua New Guinea (PNG) National Nutrition Survey were analysed to identify geographical patterns of child growth and investigate their relation to a wide range of environmental, dietary and socio economic variables. Standardized growth scores for length-for-age (LAZ), weight for-age (WAZ) and weight-for-length (WLZ) were calculated based on an internal PNG growth reference. Hierarchical Bayesian spatial models based on conditional autoregressive (CAR) priors were subsequently used to model spatial patterns in scores and their relation to different sets of covariates. The geographical differences were bigger for linear growth than for increases in weight. Growth was most reduced in parts of Milne Bay Province, Madang Province, the Torricelli/Prinz Alexander Ranges, and in the area occupied by Angan people. Socio-economic status was the most important factor determining variation in growth within populations. Differences in diet and, to a lesser extent, the physical environment were the main determining factor of differences among populations. Covariate adjustment accounted for more spatially structured variation in LAZ and WAZ than in WLZ. All variables indicating higher socio economic status were correlated with better growth, as was a high consumption of imported and local high quality foods such as cereals, legumes, tinned fish/meat or fresh fish. This indicates that nutritional interventions in PNG should aim at promoting the consumption of such high energy and high protein foods as well as strengthening the general economic base of rural populations. PMID- 11393335 TI - Short tandem repeat (STR) variation in eight village populations of the island of Korcula (Croatia). AB - The aim of this study was to analyse short tandem repeat (STR) variation using the data on nine loci (D3S1358, vWA, FGA, THO1, TPOX, CSF1PO, D5S818, D13S317, D7S820) in the populations from eight villages on the island of Korcula, Croatia, in order to analyse its genetic and population structure. The analysis of STR data in this study indicated an appreciable degree of genetic homogeneity among the studied village populations on the island, even though a so-called 'east-west dichotomy' and differentiation between the inhabitants of the most recent settlement and the remaining ones was indicated with respect to the loci CSF1PO and TPOX, respectively. The validity of STR markers in assessing genetic structure of small populations and especially in determining the relationships among geographically closely related but reproductively isolated groups remains to be further evaluated, especially in terms of a larger number of studied loci in order to possibly find specific markers useful for resolving genetic patterns of variability at regional levels. PMID- 11393336 TI - Human mitochondrial DNA sequence variation in the Moroccan population of the Souss area. AB - BACKGROUND: Various populations have contributed to the present-day gene pool of Morocco, including the autochthonous Berber population, Phoenicians, Sephardic Jews, Bedouin Arabs and sub-Saharan Africans. OBJECTIVE: The primary objective of the study was to complete a genetic description of the Berber-speaking population in the Souss region of southern Morocco, based on mitochondrial DNA (mtDNA) sequence analysis. SUBJECTS AND METHODS: The first hypervariable segment of the mtDNA control region was sequenced in a sample of 50 individuals from the Souss Valley, and the results compared with the extensive body of data available on mtDNA sequence variation in Europe and sub-Saharan Africa. RESULTS: Thirty-four different sequences were found: an estimated 68% of the sequences occurred throughout Europe, West Asia and North Africa, 26% originated in sub-Saharan Africa, and 6% belonged to the North African specific haplogroup U6. The Souss Valley mtDNA sequences indicated the presence of two populations which expanded at different times: the West Eurasian sequences in the Souss sample had a smaller average number of pairwise differences than pairs of sub-Saharan sequences. CONCLUSION: Detailed knowledge of the possible geographic origin of each sequence facilitated an interpretation of both internal diversity parameters and between population relationships. The sub-Saharan admixture in the Souss Valley matched the south-north cline of sub-Saharan influence in North Africa, also evident in the genetic distances of North African populations to Europeans and sub-Saharan Africans. PMID- 11393337 TI - D1S80 distribution in world populations with new data from the UK and the Indian sub-continent. AB - BACKGROUND: Highly polymorphic genetic markers including variable number of tandem repeats (VNTRs), amplified fragment length polymorphisms (AMP-FLPs) and short tandem repeats (STRs) have been used successfully in disease diagnostics, forensics, paternity analysis and population diversity studies. The D1S80 locus has been extensively investigated in many populations but studies on the UK and Indian subcontinent populations are limited. AIM: This study aims to enlarge our understanding of genetic variation at the D1S80 locus in the populations of the UK and the Indian subcontinent. Also, the spectrum of genetic variation at this locus in world populations is analysed. SUBJECTS AND METHODS: Six geographically and ethnically diverse populations were genotyped for the D1S80 locus using the polymerase chain reaction (PCR) technique. Two UK populations were from the East Midlands and North East England, while Brahmins, Parsis, Sinhalese and Moors represented the Indian subcontinent populations. In addition, allele frequency data of the present study were compared with 78 world populations using different methods of multivariate analyses to document level and extent of genetic diversity. RESULTS: All study populations were in Hardy-Weinberg equilibrium. A trimodal distribution (alleles 18, 24 and 31) was observed in four populations (North East England, East Midlands, Brahmins and Parsis). The Sinhalese and the Moors had different trimodal distributions. The overall heterozygosity and the level of variation are comparable to many Caucasian populations. Multivariate analyses (correspondence analysis and multidimensional scaling analysis) provided similar results in differentiation of major ethnic population groups. CONCLUSIONS: Since D1S80 variation shows considerable homogeneity within a given ethnic group, but marked variation among them, it is a useful anthropological marker for the differentiation of these populations. PMID- 11393338 TI - Growth processes leading to a large or small adult size. AB - BACKGROUND: The way in which a large size in anthropometric variables is achieved is a longstanding problem, since the pubertal spurt shows statistically and clinically little association with adult size (mostly studied for height). By analysing longitudinal growth of groups of subjects with a large or a small adult size separately for height, leg and sitting height, and bihumeral and biiliac width, we studied this problem in some detail. Of interest are growth patterns specific for these variables and for boys or girls. METHODS: The data consist of 120 boys and 112 girls followed longitudinally from 4 weeks until adulthood. Statistically, structural average velocity curves were computed for each variable and each subgroup separately for comparison. This velocity curve represents the average intensity and the average tempo of growth. Since the area under the velocity curve is adult size, differences in the growth process can be visualized. RESULTS: Both sexes show similar patterns in reaching a small or large adult size. The different variables, however, show marked differences. Only for legs is the pubertal spurt delayed for the large groups (with additional gains in prepubertal years). For sitting height and biiliac width, a slightly elevated velocity all along development (after 2 years) leads to a larger size and for bihumeral width the size of the pubertal peak is decisive. CONCLUSIONS: The steering of growth to a certain target size is qualitatively similar for boys and girls, but quite different for different anthropometric variables. This leads to questions about endocrinological control for various parts of the body and differential bone growth in development. PMID- 11393339 TI - Growth of early and late maturers. AB - BACKGROUND: This is a study on the growth of subgroups of normal children, maturing early or late, in the variables height, leg and sitting height, arm length, biiliac and bihumeral width. While a longer growth period affects adult height only marginally, less is known about the other variables. It is also of interest to see in what way a shorter growth period is compensated by a higher velocity. METHODS: Out of 120 boys and 112 girls followed from 4 weeks until adulthood, subgroups of 40 boys and 37 girls were formed with respect to the average timing (across variables) of the pubertal spurt as an indicator of maturity. RESULTS: Only leg height shows a smaller adult size for early maturers. The shorter growth period is compensated by a higher prepubertal velocity and a higher level in pubertal years. The pubertal peak is a little larger for early maturing boys but not for girls. CONCLUSIONS: There is an inherent pacemaker for growth that leads to the same adult size for a shorter growth period via a higher basic intensity. Legs are an exception since late maturers have, on average, longer legs as adults. PMID- 11393340 TI - Infants' growth charts for southern Iran. AB - Currently there are no growth charts based on local norms available for infants in Iran, and their growth is assessed by the National Centre for Health Statistics (NCHS) reference data, which is misleading. Growth charts for a cohort of 317 infants (164 girls and 153 boys) born in Shiraz (Southern Iran) in 1996 and followed for 2 years from birth are presented. All the centiles of length and weight charts are slightly above those of the NCHS charts under the age of 6 months and fall substantially below those over the age of 6 months. However, the spread is similar, so there is no suggestion that the difference is due to the prevalence of gross malnutrition. The difference shows that the use of locally based growth charts are essential for assessing the growth of children in Iran. The representativeness of our data leads us to conclude that the charts presented here are likely to be applied to the urban infant population of Iran. PMID- 11393341 TI - Weight, height, body mass index and prevalence of obesity among the adult population in Bahrain. AB - PRIMARY OBJECTIVES: To determine anthropometric measurements (weight, height, body mass index, BMI), and the prevalence of overweight and obesity based on BMI. METHODS AND PROCEDURES: A cross-sectional survey of 514 Bahraini native adults aged 30-79 years was selected from households using clustering sampling technique. OUTCOMES AND RESULTS: Findings indicate that Bahraini adults were shorter but heavier, and have higher mean BMI than their Western counterparts, suggesting a trend to obesity. The overall prevalence of overweight and obesity was 35.2% and 21.2% for men, while that for women was 31% and 48.7%, respectively. CONCLUSIONS: The results of this study confirmed the data in other Arabian Gulf countries, as obesity is a major public health problem among the adult population. Anthropometrics provided in this study can be used as baseline data for the adult population of Bahrain. PMID- 11393342 TI - A 24-week dose-titration study of the angiotensin-converting enzyme inhibitor imidapril in the treatment of mild-to-moderate essential hypertension in the elderly. AB - This multicentre, randomized, double-blind study compared the anti-hypertensive efficacy and safety of oral once-daily imidapril 5-20 mg and hydrochlorothiazide 12.5-50 mg in elderly patients with mild-to-moderate essential hypertension. After 24 weeks of treatment, there was a significant reduction in mean sitting diastolic blood pressure from 102.5 mmHg to 87.2 mmHg in the imidapril group (n = 226) and from 102.7 mmHg to 87.4 mmHg in the hydrochlorothiazide group (n = 123) (intent-to-treat population). There were corresponding reductions in sitting systolic blood pressure and standing blood pressure. At least one adverse event was reported by 46% of patients in the imidapril group and 53% of patients in the hydrochlorothiazide group. Imidapril 5-20 mg is as effective and well tolerated as hydrochlorothiazide in the treatment of mild-to-moderate hypertension in elderly patients. PMID- 11393343 TI - The significance of measuring plasma leptin in acute myocardial infarction. AB - We measured leptin concentrations in patients with acute myocardial infarction (AMI, n = 21) and in 15 age-matched controls, and compared leptin concentrations with levels of other myocardial enzymes and indicators of AMI. Blood was sampled immediately after hospital admission and at 1 h, 2 h, 3 h, 6 h and 9 h, then every 12 h until 5 days post-admission. Patients were stratified into three groups according to peak leptin concentrations: hypoleptinaemia (< 3 ng/ml); normoleptinaemia (> or = 3 - < 15 ng/ml) and hyperleptinaemia (> or = 15 ng/ml). Hypoleptinaemic AMI patients had significantly increased concentrations of plasma lactate dehydrogenase compared with normoleptinaemic patients. No significant differences in other serum markers were noted between hyperleptinaemic and normoleptinaemic AMI patients. A significant negative correlation was found between the peak concentrations of leptin and interleukin 6. Leptin may play a role in the regulation of the development of cardiac damage in patients with AMI. PMID- 11393344 TI - Effect of inhibitors of protein phosphatase 1 and 2A on erythroid colony formation: an investigation of the specificities of inhibitors. AB - To investigate the possible involvement of protein phosphatase (PP)1 and PP2A in the process of erythropoiesis, we assessed the effect of PP1 and PP2A inhibitors on erythroid colony formation using an in vitro colony formation assay. Okadaic acid (OKA), calyculin A (Cal-A) and tautomycin suppressed colony formation but 1 nor-okadaone did not. These results suggest that PP1 and PP2A both play an important role in erythropoiesis. Furthermore, higher concentrations of tautomycin were needed to suppress colony formation compared to concentrations of OKA and Cal-A. The target enzyme of inhibitors in erythropoiesis may be PP2A. PMID- 11393345 TI - Donor dendritic cells and recipient Kupffer cells in the induction of donor specific immune hyporesponsiveness. AB - The aim of this study was to investigate the ability of portovenously administered donor antigens to induce immune hyporesponsiveness. Lewis (LEW, RT 1l) rats received Brown Norway (BN, RT-1n) rat donor splenocytes, via either the portal vein (PV group) or the peripheral vein (IV group). The immune responses of LEW rats, treated with either donor BN or third party Wistar King A (WKA, RT-1k) splenocytes were established by the persistence of donor dendritic cells (DCs) in the host liver measured using fluorescence microscopy and flow cytometry and by the mixed lymphocyte reaction (MLR). The effect of intravenous gadolinium chloride (GDCl3) on the blockade of Kupffer cell function prior to portovenous administration of splenocytes was also assessed. The MLR response was strongly inhibited in a BN-restricted manner after portovenous administration of donor BN splenocytes, but not by venous nor by portovenous administration of WKA splenocytes. Immunosuppression was blocked by pretreatment with GDCl3. The percentage of donor DCs in hepatic non-parenchymal cells (NPCs) was significantly higher in the PV group compared with the IV group. Treatment with GDCl3 decreased the percentage of donor DCs. In addition, cytotoxic T lymphocyte antigen 4 (CTLA4/CD152), which may function as an immune attenuator, was strongly stained, and B7 was weakly stained in recipient liver in the PV group compared with the IV group. These results suggest that both donor DCs and recipient Kupffer cells (self DCs) are involved in the induction of immune hyporesponsiveness by donor cells. This occurs via portovenous administration, in which a signal of the CTLA4 B7 pathway played an important part in inhibiting the interaction of CD28 and its B7 ligands. PMID- 11393346 TI - Effects of telmisartan on renal excretory function in conscious dogs. AB - The present study investigated the effects of telmisartan, a selective AT1 receptor antagonist, on renal function in dogs. Conscious female dogs were treated with (i) vehicle (controls) and three doses of telmisartan (0.03 mg/kg, 0.1 mg/kg and 0.3 mg/kg) administered intravenously; (ii) vehicle and three doses of telmisartan (0.3 mg/kg, 1.0 mg/kg and 3.0 mg/kg) administered orally; or (iii) 1.0 mg/kg per day telmisartan orally for 12 days. Eight dogs were used for each experiment. Each of the four treatments in (i) and (ii) was administered 7 days apart. During the 6 h after intravenous administration, urine volume was significantly higher in dogs treated with telmisartan 0.1 mg/kg (8.5 +/- 1.6 ml/kg) and 0.3 mg/kg (7.0 +/- 0.9 ml/kg) than controls (2.7 +/- 0.3 ml/kg; P < 0.05), and renal sodium excretion was increased significantly with telmisartan 0.03 mg/kg (803 +/- 124 micromol/kg), 0.1 mg/kg (1039 +/- 213 micromol/kg) and 0.3 mg/kg (966 +/- 161 micromol/kg) versus controls (159 +/- 21 micromol/kg; P < 0.05). Oral telmisartan at doses of 1.0 mg/kg and 3.0 mg/kg also produced significant increases in urine volume (7.2 +/- 1.1 ml/kg and 6.6 +/- 1.2 ml/kg, respectively) and renal sodium excretion (599 +/- 146 micromol/kg and 555 +/- 131 micromol/kg, respectively) compared with controls (2.8 +/- 0.5 ml/kg and 80 +/- 33 mciromol/kg; P < 0.05) over the 6-h post-dose period. Telmisartan at all intravenous doses and at 3.0 mg/kg orally increased the urinary excretion of chloride significantly over the 6-h post-dose period compared with vehicle alone. The excretion of potassium and creatinine were unchanged by any treatment. Telmisartan 1.0 mg/kg administered orally for 12 days produced similar results. In conclusion, acute intravenous or oral as well as subchronic oral administration of telmisartan to conscious dogs promotes diuresis and natriuresis without affecting potassium or creatinine excretion. PMID- 11393347 TI - Treatment of acute myeloblastic leukaemia in a patient with Bombay blood type: a case report. AB - A 62-year-old female was admitted to our hospital with suspected acute leukaemia and after investigation we diagnosed acute myeloblastic leukaemia (AML-M1). The patient's blood type was found to be the very rare Bombay type and surveillance of her relatives showed the same blood type in her male cousin on her mother's side. Alongside chemotherapy the patient received 4000 ml of frozen Bombay-type red cells, 1400 ml of concentrated red cells in manitol adenine phosphate solutions and 360 units of type O concentrated platelets without marked effects. The anti-H antibody was initially at 128 dilution but for unknown reasons increased to 2048 dilution after remission of AML-M1. About 3 months after hospitalization the patient died of Cryptococcus neoformans pneumonia despite strict precautions against infection. Although AML-M1 is a common adult leukaemia and is chemosensitive to anti-leukaemic drugs, neither AML-M1 in a patient with Bombay-type red cells nor its treatment with chemotherapy and transfusion with type Oh frozen red cells have previously been reported. PMID- 11393348 TI - Chronic obstructive pulmonary disease. PMID- 11393349 TI - Clinical experience in Germany of treating community-acquired respiratory infections with the new 8-methoxyfluoroquinolone, moxifloxacin. AB - Moxifloxacin, a new 8-methoxyfluoroquinolone, was evaluated in a large community based study involving 16,007 patients over a 9-month period. This study was designed as a large post-marketing observational study of the speed, efficacy and tolerability of moxifloxacin when used in clinical practice for the treatment of community-acquired bacterial pneumonia, or acute exacerbations of chronic bronchitis. Physicians and patients were specifically questioned about overall efficacy and safety as well as symptom relief. According to physicians' assessments 96.3% of patients were cured or improved after moxifloxacin treatment. Symptom relief ('feeling better') occurred in almost 70% of patients by day 3 and only 2.3% reported an adverse drug reaction. No individual adverse drug reaction was reported at a frequency above 1%. Among the 209 events considered as serious, only 34 were regarded as possibly or probably related to therapy. There were no moxifloxacin-related clinically relevant cases of phototoxicity, hepatotoxicity or cardiotoxicity. Overall, 92.1% of patients considered moxifloxacin to have been beneficial. PMID- 11393350 TI - Moxifloxacin in acute exacerbations of chronic bronchitis: clinical evaluation and assessment by patients. AB - The clinical success of a 5-day course of oral moxifloxacin (administered once daily at a dose of 400 mg) was evaluated in 328 patients with acute exacerbations of chronic bronchitis (Anthonisen type 1) in a non-comparative study conducted by chest physicians in private practice. Results were assessed on the basis of clinical parameters and, for the first time in a trial involving oral moxifloxacin, by the surrogate marker of patient satisfaction. Improvement in (and severity of) cough, dyspnoea, chest pain and sputum were scored daily by patients. Cough, chest pain and purulent sputum production improved rapidly within the first 5 days of treatment. At least 90% of patients were satisfied with the antibiotic. The clinical success rate (cure and improvement) for all patients involved (intent-to-treat analysis) was 90.5%. The most commonly experienced adverse events were gastrointestinal related, with diarrhoea the most frequent of these (2.7% of all patients). PMID- 11393352 TI - Clinical practice and future needs in recombinant human granulocyte colony stimulating factor treatment: a review of randomized trials in clinical haemato oncology. AB - Recombinant human granulocyte colony-stimulating factor (rHuG-CSF) may have a significant impact on preventing infections associated with chemotherapy-induced neutropenia, as well as shortening time to tree lineage engraftment following high-dose chemotherapy and progenitor transplantation. However, the scientific literature documenting evidence-based practice is insufficient and often misinterpreted. This review presents data and discusses the evidence for actual clinical practice in the use of rHuG-CSF in conventional cyclic chemotherapy, either prophylactic or therapeutic, and high-dose therapy, either in priming for mobilization or post-transplantation. In the past decade, many reports have based their conclusions on surrogate markers, and it is time to move towards evaluation of clinically relevant factors. Data must be generated prospectively based on current clinical practice, and several issues must be considered and evaluated to define the true clinical benefit of rHuG-CSF with or without stem-cell support. Evaluation should include complications and needs for resources as well as impact on toxicity and efficacy of conventional or high-dose chemotherapy. PMID- 11393351 TI - Treatment outcomes in acute exacerbations of chronic bronchitis: comparison of macrolides and moxifloxacin from the patient perspective. AB - Moxifloxacin, a new respiratory quinolone, was compared with the macrolides azithromycin, clarithromycin and roxithromycin in a cohort study to assess clinical, safety and health-related outcomes of these antimicrobials in general practice settings. In total 332 patients with acute exacerbations of chronic bronchitis (AECB) each received one of the antimicrobial agents for a standard short course of therapy. Random allocation of therapeutic agents occurred by centre, not individuals, and the drugs were prescribed in an open manner. In addition to clinical evaluation by their physicians, all patients kept daily diaries to assess AECB symptoms over the study period, therapy received and quality of life. The overall clinical response rate was 96% and all four regimens were well tolerated. After 14 days there were no significant differences between the study groups, but analyses of patients' daily evaluations of certain AECB specific symptoms showed a faster response rate in the moxifloxacin group. PMID- 11393353 TI - Oxidative lime pretreatment of high-lignin biomass: poplar wood and newspaper. AB - Lime (Ca[OH]2) and oxygen (O2) were used to enhance the enzymatic digestibility of two kinds of high-lignin biomass: poplar wood and newspaper. The recommended pretreatment conditions for poplar wood are 150 degrees C, 6 h, 0.1 g of Ca(OH)2/g of dry biomass, 9 mL of water/g of dry biomass, 14.0 bar absolute oxygen, and a particle size of -10 mesh. Under these conditions, the 3-d reducing sugar yield of poplar wood using a cellulase loading of 5 filter paper units (FPU)/g of raw dry biomass increased from 62 to 565 mg of eq. glucose/g of raw dry biomass, and the 3-d total sugar (glucose + xylose) conversion increased from 6 to 77% of raw total sugars. At high cellulase loadings (e.g., 75 FPU/g of raw dry biomass), the 3-d total sugar conversion reached 97%. In a trial run with newspaper, using conditions of 140 degrees C, 3 h, 0.3 g of Ca(OH)2/g of dry biomass, 16 mL of water/g of dry biomass, and 7.1 bar absolute oxygen, the 3-d reducing sugar yield using a cellulase loading of 5 FPU/g of raw dry biomass increased from 240 to 565 mg of eq. glucose/g of raw dry biomass. A material balance study on poplar wood shows that oxidative lime pretreatment solubilized 38% of total biomass, including 78% of lignin and 49% of xylan; no glucan was removed. Ash increased because calcium was incorporated into biomass during the pretreatment. After oxidative lime pretreatment, about 21% of added lime could be recovered by CO2 carbonation. PMID- 11393354 TI - Adsorption of Thermomonospora fusca E5 and Trichoderma reesei cellobiohydrolase I cellulases on synthetic surfaces. AB - The interfacial behavior of Thermomonosporafusca E5 and Trichoderma reesei cellobiohydrolase I (CBHI) cellulases were studied at synthetic surfaces. For this purpose, colloidal silica and polystyrene particles were used to prepare cellulase-particle suspensions that could be analyzed by solution-phase techniques. Circular dichroism spectroscopy of each cellulase, alone as well as in suspension with silica, was used to determine whether structural changes occurred on adsorption. Changes in spectra were observed for CBHI, but not for E5. Gel-permeation chromatography of the cellulase-particle suspensions showed that neither cellulase binds to silica, suggesting that changes in spectra for CBHI were a result of solution-phase phenomena. Microfiltration of cellulase polystyrene suspensions showed that both cellulases bind to polystyrene. However, circular dichroism experiments with polysterene proved unworkable, owing to excessive light absorption by the polystyrene. Adsorption kinetics of each cellulase were recorded, in situ, at hydrophilic and silanized, hydrophobic silica surfaces using ellipsometry. Ellipsometric data recorded for each cellulase at hydrophilic silica showed insignificant adsorption. Binding did occur between each cellulase and silanized silica, most likely mediated through hydrophobic associations. Adsorption in this case was irreversible to dilution. PMID- 11393355 TI - Fermentation and costs of fuel ethanol from corn with Quick-Germ process. AB - The Quick-Germ process developed at the University of Illinois at Urbana Champaign is a way to obtain corn oil, but with lower capital costs than the traditional wet-milling process. Quick-Germ has the potential to increase the coproduct credits and profitability of the existing dry-grind fuel ethanol process, but the fermentability of the corn remaining after oil recovery has not been tested. Therefore, a series of pilot scale (50 L) fermentations was carefully controlled and monitored with unique methods for standard inoculation and automatic sampling. It was found that the concentration of suspended solids was significantly reduced in the Quick-Germ fermentations. When compared at the same concentration of fermentable sugars, the fermentation rate and yield were not statistically different from controls. When Quick-Germ was integrated into a state-of-the-art dry-grind fuel ethanol process, computer simulation and cost models indicated savings of approx $0.01/L of ethanol ($0.04/gal) with the Quick Germ process. Additional savings associated with the lower suspended solids could not be quantified and were not included. However, the savings are sensitive to the price of corn oil. PMID- 11393356 TI - Metabolic flux analysis of Clostridium thermosuccinogenes: effects of pH and culture redox potential. AB - Clostridium thermosuccinogenes are anaerobic thermophilic bacteria that ferment various carbohydrates to succinate and acetate as major products and formate, lactate, and ethanol as minor products. Metabolic carbon flux analysis was used to evaluate the effect of pH and redox potential on the batch fermentation of C. thermosuccinogenes. In a first study, the effects of four pH values (6.50, 6.75, 7.00, and 7.25) on intracellular carbon flux at a constant redox potential of 275 mV were compared. The flux of carbon toward succinate and formate increased whereas the flux to lactate decreased significantly with a pH increase from 6.50 to 7.25. Both specific growth rate and specific rate of glucose consumption were unaffected by changes in pH. The fraction of carbon flux at the phosphoenolpyruvate (PEP) node flowing to oxaloacetate increased with an increase in pH. At the pyruvate node, the fraction of flux to formate increased with increasing pH. At the acetyl CoA node, the fraction of flux to acetate increased significantly with an increase in pH. A second study elucidated the effect of four controlled culture redox potentials (-225, -250, -275, and -310 mV) on metabolic carbon flux at a constant pH of 7.25. Lower values of culture redox potential were correlated with increased succinate, acetate, and formate fluxes and decreased ethanol and hydrogen fluxes in C. thermosuccinogenes. Lactate formation was not significantly influenced by redox potential. At the PEP node, the fraction of carbon to oxaloacetate increased with a decrease in redox potential. At the pyruvate node, the fraction of carbon to formate increased, while at the acetyl CoA node, the fraction of carbon flux to acetate increased with reduced redox potential. The presence of hydrogen in the headspace or the addition of nicotinic acid to the growth media resulted in increased hydrogen and ethanol fluxes and decreased succinate, acetate, formate, and lactate fluxes. PMID- 11393357 TI - Sugar-mediated crosslinking of alpha-biotinylated-Lys to cysteamine-agarose support: a method to isolate Maillard Lys-Lys-like crosslinks. AB - Advanced glycation end products (AGEs) and, specifically, protein-protein AGE crosslinks have long been studied for their potential role in aging, diabetic complications and Alzheimer disease. With few exceptions, the chemical nature of these structures remains unknown. We report here a simple approach that allows the preparation and isolation of milligram quantities of sugar-mediated AGE Lys Lys-like crosslinks from glycation mixtures. The method is based on a sugar dependent incorporation of N(alpha)-biotinyl-L-Lys into cysteaminyldisulfide Sepharose 6B (AE-S-S-Sepharose 6B). Glycation mixtures with six different sugars showed a time- and sugar-dependent decrease in the concentration of the support bound primary amino groups and accounted for almost 90% loss of cysteaminyl amino groups at the end of the various incubation periods. 4-Hydroxyazobenzene-2 carboxylic acid-avidin assays indicated the incorporation of N(alpha)-biotinyl-L Lys equal to 8% of the total support amino groups with methylglyoxal after 7 d and 1% with fructose and glucose after 1 mo of incubation. Treatment of the washed, sugar-modified supports with 2-mercaptoethanol released the bulk of the bound AGE modifications and the crosslinks. Subsequent fractionation of these preparations over a monomeric avidin column afforded a complete separation of sugar-mediated AGE modifications and the crosslinks. Depending on the sugar employed, micromolar amounts of biotinylated Lys-Lys-like crosslinks were generated by this two-step procedure from 8 mL of the original AE-S-S-Sepharose 6B. PMID- 11393358 TI - Veterinarians join emergency eradication effort. PMID- 11393359 TI - Differing opinions of raw food diet research. PMID- 11393360 TI - Differing opinions of raw food diet research. PMID- 11393361 TI - Differing opinions of raw food diet research. PMID- 11393363 TI - New perspectives on transmission of foodborne pathogens and antimicrobial resistance. PMID- 11393362 TI - Renal failure associated with ingestion of grapes or raisins in dogs. PMID- 11393364 TI - ECG of the month. Atrial fibrillation. PMID- 11393365 TI - What is the most likely cause for the poor pregnancy rate and the distribution for stage of pregnancy in the histogram? PMID- 11393366 TI - Vision and hearing in horses. PMID- 11393367 TI - West Nile virus encephalomyelitis in eight horses. PMID- 11393368 TI - Effects of hydrochlorothiazide and diet in dogs with calcium oxalate urolithiasis. AB - OBJECTIVE: To determine whether hydrochlorothiazide (HCTZ) reduces urinary calcium excretion in dogs with calcium oxalate urolithiasis. DESIGN: Original study. ANIMALS: 8 dogs with calcium oxalate urolithiasis. PROCEDURE: 4 treatment protocols were evaluated in each dog (a low calcium, low protein diet designed to prevent calcium oxalate urolith formation with and without administration of HCTZ [2 mg/kg (0.9 mg/lb) of body weight, PO, q 12 h] and a maintenance diet with higher quantities of protein and calcium with and without administration of HCTZ). At the end of each 2-week treatment period, 24-hour urine samples were collected. Blood samples were collected during the midpoint of each urine collection period. Analysis of variance was performed to evaluate the effects of HCTZ and diet on urine and serum analytes. RESULTS: Hydrochlorothiazide significantly decreased urine calcium and potassium concentration and excretion. Hydrochlorothiazide also significantly decreased serum potassium concentration. Compared with the maintenance diet, the urolith prevention diet significantly decreased urine calcium and oxalic acid concentration and excretion. Dogs consuming the urolith prevention diet had significantly lower serum concentrations of albumin and urea nitrogen. CONCLUSIONS AND CLINICAL RELEVANCE: Administration of HCTZ decreased urine calcium excretion in dogs with a history of calcium oxalate urolith formation. The greatest reduction in urine calcium concentration and excretion was achieved when dogs received HCTZ and the urolith prevention diet. Results of this study suggest that the hypocalciuric effect of HCTZ will minimize recurrence of calcium oxalate urolith formation in dogs; however, long-term controlled clinical trials are needed to confirm the safety and effectiveness of HCTZ. PMID- 11393369 TI - Hyperactivity and alopecia associated with ingestion of valproic acid in a cat. AB - A 1-year-old castrated male cat was evaluated because of alopecia of approximately 4 to 5 months' duration as well as hyperactive behavior. It was later determined that the cat was ingesting valproic acid by eating food to which it had been added for daily administration to a child in the household who had cerebral palsy. The clinical signs slowly resolved after the source of valproic acid was removed. This emphasizes the sensitivity of cats to drugs that are commonly used in humans. It was not determined whether the clinical signs that developed in this cat were caused by an adverse reaction or from toxicosis as a result of prolonged hepatic elimination of valproic acid, which requires glucuronide metabolism for disposition. However, the cat recovered completely following removal of the drug and prevention of further exposure. This report emphasizes the importance of obtaining a careful and complete history from the owner regarding an animal and its environment. In the cat of this report, the owner had not considered the impact of the presence of the drug in the child's food. PMID- 11393370 TI - Hyponatremia and hyperkalemia associated with peritoneal effusion in four cats. AB - Four cats with considerable peritoneal effusion and corresponding hyponatremia and hyperkalemia were evaluated. The Na:K ratio in all cats was < 25, which is suggestive of adrenal insufficiency. An ACTH stimulation test was performed on 3 cats for evaluation of adrenal gland function. Serum cortisol and aldosterone concentrations did not support a diagnosis of adrenal gland insufficiency. In 1 cat, histologic evaluation of the adrenal glands at necropsy also failed to support a diagnosis of hypoadrenocorticism. On the basis of these findings, and because hyponatremia and hyperkalemia could not be readily explained by another cause, the electrolyte abnormalities were presumed to be secondary to peritoneal effusion. PMID- 11393371 TI - Iatrogenic copper deficiency associated with long-term copper chelation for treatment of copper storage disease in a Bedlington Terrier. AB - A 9-year-old Bedlington Terrier was evaluated because of weight loss, inappetence, and hematemesis. Copper storage disease had been diagnosed previously on the basis of high hepatic copper concentration. Treatment had included dietary copper restriction and administration of trientine for chelation of copper. A CBC revealed microcytic hypochromic anemia. High serum activities of liver enzymes, high bile acid concentrations, and low BUN and albumin concentrations were detected. Vomiting resolved temporarily with treatment, but the clinicopathologic abnormalities persisted. Results of transcolonic portal scintigraphy suggested an abnormal shunt fraction. Results of liver biopsy and copper quantification revealed glycogen accumulation and extremely low hepatic copper concentration. Serum and hair copper concentrations were also low. Chelation and dietary copper restriction were tapered and discontinued. Clinical signs and all clinicopathologic abnormalities improved during a period of several months. PMID- 11393372 TI - Ventilatory failure, ventilator management, and outcome in dogs with cervical spinal disorders: 14 cases (1991-1999). AB - OBJECTIVE: To determine the incidence of and risk factors for ventilatory failure in dogs undergoing surgery for treatment of cervical spinal disorders and to document ventilator management, clinical course, and long-term outcome of dogs that experienced ventilatory failure in association with cervical spinal disorders or their management. DESIGN: Retrospective study. ANIMALS: 14 dogs. PROCEDURE: Dogs with cervical spinal disorders that required positive-pressure ventilation (PPV) were identified, and signalment, concurrent diseases, neurologic status at initial examination, clinical course, pulmonary function before, during, and after PPV, management techniques, complications, and outcome were recorded. Dogs that underwent surgery and required PPV were compared with dogs that underwent cervical spinal surgery during the same period that did not require PPV. RESULTS: 14 dogs with cervical spinal disorders required PPV to treat hypoventilation, including 13 of 263 (4.9%) dogs that underwent surgery for cervical spinal disorders. Lesions between the second and fourth cervical vertebrae and treatment by means of a dorsal decompressive laminectomy were associated with a significantly increased risk of perioperative hypoventilation. Pulmonary gas exchange function was normal or nearly normal throughout the course of PPV in dogs that survived. Ten dogs survived, and 9 of the 10 regained neurologic function. All 9 dogs that regained neurologic function had deep pain perception on initial examination at the veterinary teaching hospital. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that a small percentage of dogs with cervical spinal disorders may require perioperative ventilatory support. With prolonged PPV and aggressive management, a good outcome may be achieved in dogs similar to those described in the present study. PMID- 11393373 TI - Survival rates for horses undergoing stapled small intestinal anastomosis: 84 cases (1988-1997). AB - OBJECTIVE: To determine whether location and type of small intestinal anastomosis and other variables were associated with short- and long-term survival rates in horses undergoing stapled small intestinal anastomosis. DESIGN: Retrospective study. ANIMALS: 84 horses that underwent small intestinal anastomosis. PROCEDURE: Medical records from 1988 to 1997 were examined for horses that underwent stapled small intestinal anastomosis. Horses were allotted into 4 groups: jejunojejunostomy (n = 27), jejunoileostomy (11), jejunoileocecostomy with small intestinal resection (20), and jejunoileocecostomy without small intestinal resection (26). Survival rates and other variables were determined. RESULTS: Horses that underwent jejunoileocecostomy without resection had a significantly higher survival rate at all intervals than did horses in the other 3 groups. At 180 and 365 days after surgery, horses that underwent jejunojejunostomy had a significantly lower survival rate than those that underwent jejunoileocecostomy with intestinal resection. Horses that underwent jejunoileocecostomy with intestinal resection had a significantly higher survival rate at 1 year, compared with the combined jejunojejunostomy and jejunoileostomy groups. Preoperative heart rate was inversely associated with survival rate. Overall survival rates at discharge and 1 year were 81 and 56%, respectively. For horses that underwent small intestinal resection, survival rates at discharge and 1 year were 65 and 47%, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: Horses that underwent anastomosis of the small intestine to the cecum without resection had the highest survival rate, compared with horses that required intestinal resection. In some instances, resection and anastomosis involving the cecum had better prognosis than resections involving 2 segments of the small intestine. PMID- 11393374 TI - Bacterial isolates from blood and their susceptibility patterns in critically ill foals: 543 cases (1991-1998). AB - OBJECTIVE: To assess microorganisms isolated from blood specimens obtained from critically ill neonatal foals and to evaluate their antimicrobial susceptibility patterns. DESIGN: Retrospective study. ANIMALS: 543 neonatal foals. PROCEDURE: Medical records of foals that were < 1 month old and were admitted to a referral neonatal intensive care unit were reviewed for results of bacteriologic culture of blood and antimicrobial susceptibility patterns. RESULTS: At least 1 microorganism was isolated from 155 of 543 (28.5%) foals. Escherichia coli was the most commonly isolated bacterium. A single gram-positive organism was detected in 49 foals. Although 90% of the E coli isolates were susceptible to amikacin, some gram-negative and gram-positive organisms had resistance against multiple antimicrobials. CONCLUSIONS AND CLINICAL RELEVANCE: Gram-negative bacteria remain the most common isolates from blood of neonatal foals; however, gram-positive organisms were also found, and with greater prevalence than reported elsewhere. Susceptibility patterns may vary, and resistance to multiple antimicrobials may develop. This is especially true for organisms such as Enterobacter spp and Enterococcus spp. Prudent empirical treatment for neonatal sepsis should include broad-spectrum antimicrobials. PMID- 11393375 TI - Effect of lameness on the calving-to-conception interval in dairy cows. AB - OBJECTIVE: To examine the relationship between lameness and the duration of the interval from calving to subsequent conception in lactating dairy cows. DESIGN: Cohort study. ANIMALS: 837 dairy cows. PROCEDURE: Cows affected with lameness were classified into 1 of 4 groups on the basis of types of disease or lesions observed, including foot rot, papillomatous digital dermatitis, claw lesions, or multiple lesions. Cows not affected with lameness were classified as healthy. Time from calving to conception was compared between lame cows and healthy cows. RESULTS: 254 (30%) cows were affected with lameness during lactation. Most lame cows (59%) had claw lesions. Lame cows with claw lesions were 0.52 times as likely to conceive as healthy cows. Median time to conception was 40 days longer in lame cows with claw lesions, compared with healthy cows. Number of breedings per conception for lame cows with claw lesions was significantly higher than that for healthy cows. CONCLUSION AND CLINICAL RELEVANCE: Claw lesions were the most important cause of lameness, impairing reproductive performance in dairy cows, as indicated by a higher incidence of affected cows and a greater time from calving to conception and a higher number of breedings required per conception, compared with healthy cows. PMID- 11393376 TI - Management strategies to decrease the prevalence of mastitis caused by one strain of Staphylococcus aureus in a dairy herd. AB - The dairy herd at Washington State University had an outbreak of mastitis caused by a single strain of Staphylococcus aureus. The outbreak strain, termed novel, could not be controlled with routine contagious mastitis pathogen control procedures (incidence, 3.4 infections/100 cow months; peak prevalence > 22%). Our objective was to implement mastitis control measures that would decrease the incidence and prevalence of intramammary infection (IMI) caused by S aureus in the herd. The following intervention strategies were successfully implemented: strict segregation of cattle with IMI caused by S aureus, intensified culling of cattle with multiple-quarter IMI caused by S aureus, and inducing cessation of lactation of infected quarters in single-mammary-quarter infected cattle. One year after implementation of these control measures, incidence of IMI caused by S aureus was 0.35 infections/100 cow months, and prevalence had decreased from 20 to 8%. PMID- 11393377 TI - Otitis media in a llama. AB - A 7-month-old female llama was examined because of chronic otitis media and externa of 7 months' duration. Radiographically, the tympanic bullae appeared thicker than normal, and the ventral borders were poorly defined; the left external acoustic meatus (ear canal) appeared to be narrower than the right. The llama was treated with penicillin, and the ear canals were lavaged daily. Contrast radiography was performed on day 15 to determine the shape and size of the ear canals and evaluate the integrity of the tympanic membranes. Contrast medium was visible radiographically in the left tympanic bulla, indicating that the left tympanic membrane was ruptured, but the right tympanic membrane appeared to be intact. The left ear canal was narrower than the right, and the bony ear canals had a well-defined sigmoid shape. The right ear improved with medial treatment alone, but the left ear did not. Therefore, lateral ear canal resection was performed. After surgery, however, exudate was still evident in the left ear canal, and the llama became more lethargic and more reluctant to eat. Lateral bulla osteotomy was attempted, but no purulent material was obtained, and curettage of the bulla resulted in hemorrhage. Because of this and because of the llama's poor physical condition, a decision was made to euthanatize the llama. The sigmoid shape of the bony ear canal and the multicompartmental nature of the tympanic bulla make surgical treatment of otitis media and externa in llamas difficult. Further study of surgical treatments for otitis media in llamas is needed. PMID- 11393378 TI - Alternative treatments for focal and segmental glomerular sclerosis. AB - Corticosteroids represent the first therapeutic approach for patients with focal and segmantal glomerulosclerosis. What to do in patients who do not respond to corticostetoids is still uncertain. Cytotoxic agents have been tried. The results are usually poor when these drugs are given for short periods but almost half of patients may enter remission ir treatment is prolonged. Some decrease in proteinuria may be obtained with mycophenolav, mofetil but the available reports are still scanty and the follow ups are short. Cyclosporine has been largely used both in controlled and non controlled trials with favorable results. However, being the drug nephrotoxic caution should be recommended with its use. Plasma exchange and lipopheresis may attempted in resistant cases. PMID- 11393379 TI - Benign nephrosclerosis: incidence, morphology and prognosis. AB - AIMS: Study of benign nephrosclerosis (BNS) is often mixed up with IgA nephritis (IgAN) associated with hypertension or thin basement membrane disease (TBMD). Here we examined the clinicopathological features, incidences and prognosis of decompensated BNS. MATERIALS AND METHODS: BNS was identified in 590 (8.3%) adult cases among 7,108 renal biopsies of a mean age of 56.5 years (male: female ratio = 2.5:1). The post-biopsy follow-up period ranged from 3 to 22 years (10.1 +/- 4.6 years). RESULTS: Patients with progressive BNS were more likely to develop end-stage renal disease within 5 years of biopsy. Poor prognostic factors included poor or no control of arterial blood pressure by anti-hypertensive drugs, global glomerulosclerosis (GS) (> or = 41%) at biopsy, presence of collapsed glomeruli and/or segmented or semi-global GS. Tubulointerstitial damage, glomerular hypertrophy and loop dilatation were secondary to GS. Gender, duration of HT and onset of HT to biopsy were not significant factors. CONCLUSION: GS in BNS is due to ischemia induced by luminal narrowing or obstruction of preglomerular vessels, and glomerular HT due to loss of autoregulation in preglomerular vessels (irregularly shaped atrophic or segmented medial smooth muscle cells, with expansion of extracellular matrix with or without fibrous intimal thickening). GS resulted in luminal dilatation. Both pathological changes affecting the glomerulus may occur in the same kidney and different nephron units. PMID- 11393380 TI - Urinary type IV collagen excretion reflects renal morphological alterations and type IV collagen expression in patients with type 2 diabetes mellitus. AB - AIM: We tried to establish the significance of quantifying urinary type IV collagen (IV-C) excretion for the evaluation of renal involvement of type 2 diabetic patients. METHODS: Twenty patients (13 males and 7 females; age range 31 to 69 years) with type 2 diabetes mellitus had undergone renal biopsy and relationship between the severity of morphological alteration, IV-C expression and urinary IV-C excretion were examined. RESULTS: Urinary IV-C excretion significantly correlated with mesangial expansion score (p = 0.49, p < 0.05) and tubulointerstitial injury score (p = 0.56, p < 0.05). Furthermore, urinary IV-C excretion significantly correlated with both glomerular (r = 0.56, p < 0.01) and tubulointerstitial IV-C expression areas. Urinary protein excretion also correlated with mesangial expansion score and tubulointerstitial injury score. However, it did not correlate with the expression of IV-C in the kidney. CONCLUSIONS: These results suggest that urinary IV-C excretion reflects the pathogenetic process of diabetic nephropathy, which urinary protein excretion alone cannot do sufficiently. It can be concluded that urinary IV-C excretion could be a more useful marker for the evaluation of renal involvement of type 2 diabetic patients. PMID- 11393381 TI - C-reactive protein frequently localizes in the kidney in glomerular diseases. AB - AIM: In recent reports, C-reactive protein (CRP) has emerged as a component that may play important roles in atherogenesis. Based on the analogies set out in a previous report between focal-segmental sclerosis and atherosclerosis, we hypothesized that CRP contributes to the pathogenesis of glomerular diseases. To our knowledge, no immunohistochemical study of CRP localization in the kidneys has been previously reported. PATIENTS AND METHODS: In the present study, we investigated 106 kidney biopsy specimens from children with various types of glomerular diseases and minor glomerular abnormalities. Of the 106 cases, 74 were proliferative diseases, 17 were non-proliferative diseases, and 15 were minimal change nephrotic syndrome (MCNS). Immunohistochemical staining was performed using monoclonal antibody to CRP. RESULTS: CRP immunoreactivity was found in 48 of 106 (45.3%) specimens. CRP deposition was encountered more often in patients with proliferative diseases (56.8%) than in those with non-proliferative diseases (23.5%) (p < 0.01). CRP deposition, most frequently observed along the capillary walls of glomeruli, was found in 33 of 46 (71.7%) cases with positive expression of CRP. CRP was also located in the peritubular capillary walls and small vessels in the interstitium in 13 of 46 cases (28.3%). CRP deposition was also found in 2 of 15 cases of MCNS. The two MCNS specimens showing positive CRP immunoreactivity were both from patients who had undergone cyclosporin therapy. CRP deposition was not shown in any cases treated with steroids or cyclophosphamide. The cases of patients who had undergone renal biopsies within 6 months after onset revealed a tendency toward positive CRP deposition. The clinical outcomes at the latest follow-up were quite similar between the groups of patients with and without CRP deposition. CONCLUSIONS: We surmise that circulating CRP may deposit at the site of endothelial injury, and may not be relevant to the progression of renal lesions. PMID- 11393382 TI - Clinical characterization of Dobrava hantavirus infections in Germany. AB - There is increasing evidence that Dobrava (DOBV) but not Hantaan (HTNV) hantavirus is a hemorrhagic fever with renal syndrome (HFRS) causing agent in Central Europe. However, only single clinical cases of HFRS due to acute DOBV infection have been described so far. We report on three male patients from a non endemic hantavirus focus in Northern Germany who suffered from mild to moderate HFRS strongly resembling nephropathia epidemica. Serotyping by detection of hantavirus species-specific neutralizing antibodies revealed acute infections by the HTNV-related hantavirus DOBV in all three cases. Since DOBV infections in the Balkans frequently present as severe HFRS, our cases suggest that Central European DOBV infections have a different, less severe clinical outcome. These differences in DOBV virulence towards humans might be due to the existence of different genetic lineages of DOBV. PMID- 11393383 TI - A randomized and double-blind comparison of isradipine and spirapril as monotherapy and in combination on the decline in renal function in patients with chronic renal failure and hypertension. AB - AIMS: Treatment of hypertension in patients with chronic renal failure has been shown to postpone the decline in renal function. Treatment with an ACE inhibitor has been shown to be superior to conventional antihypertensive treatment, but it is not known how an ACE inhibitor compares to treatment with a calcium channel blocker or to treatment with a combination of these drugs. The aim of the study was to evaluate the rate of decline in GFR in patients with chronic renal failure and hypertension treated with isradipine and spirapril as monotherapy and in combination. METHODS: Sixty patients with chronic renal failure and hypertension were enrolled in the study. After enrollment, patients were followed prospectively for 6 months in the outpatient clinic on their usual antihypertensive medication, and then randomized to a double-blinded comparison of either spirapril 6 mg daily, isradipine 5 mg daily or spirapril 3 mg and isradipine 2.5 mg daily. After randomization, patients were followed for 21 months or until the need for dialysis. Every 3 months before and 3.5 months after randomization the glomerular filtration rate was measured by 51Cr-EDTA clearance and the effective renal plasma flow evaluated using the renal clearance of paraaminohippuric acid. RESULTS: Blood pressure and the decline in glomerular filtration rate did not differ between the groups before randomization. After randomization, the mean decline in the glomerular filtration rate was -0.32 ml/(min x month x 1.73 m2) in the spirapril group, -0.58 ml/(min x month x 1.73 m2) in the isradipine group and -0.14 ml/(min x month x 1.73 m2) in the combination group (p = 0.38). Twelve patients, 4 in each group, reached end-stage renal failure. No significant difference was found with respect to diastolic (p = 0.10) or systolic blood pressure (p = 0.08) during the treatment period, but a trend towards a better blood pressure control in the combination group was present. During treatment, the rate of decline in renal plasma flow did not differ significantly between the groups (p = 0.09), neither did the changes in filtration fraction (FF) (p = 0.58) nor the mean FF (p = 0.22) during the treatment. CONCLUSIONS: Our study indicated differences between the 3 treatment modalities in favor of combined therapy with respect to both the rate of decline in GFR and blood pressure control, but the differences where insignificant. Thus, the treatments might differ, but we were unable to confirm this because of large variation in GFR and small sample size. PMID- 11393384 TI - Urine analysis performed by flow cytometry: reference range determination and comparison to morphological findings, dipstick chemistry and bacterial culture results--a multicenter study. AB - AIM: To validate whether quantitative flow-cytometric analysis of particulate matter in urine would allow for accurate and rapid enumeration of red blood cells (RBC), leukocytes (WBC), squamous epithelial cells (EC), casts, and bacteria, a Sysmex UF-100 analyzer was tested in a multicenter study. MATERIAL AND METHODS: At first, reference values were established and found to be < 14 for RBC, < 16 for WBC, < 9 for EC, < 2 for casts and < 173 for bacteria, respectively (counts per microl; 97.5 percentile). Due to the wide use of dipstick and microscopic sediment analysis in routine urine diagnostics, comparative studies on 950 random urine samples were performed. Bacterial counting combined with WBC enumeration was further compared in 266 routine urinary microbiologic cultures. RESULTS: Good correlations were found comparing UF-100 results of RBC (r = 0.89), WBC (r = 0.94), and EC (r = 0.74) with Fuchs-Rosenthal Chamber (FRC) counts. However, some misclassification of casts (r = 0.32) could be observed. Correlations of UF-100 with dipstick and sediment testing was significant (p < 0.001), but the scatter of the latter two methods is too wide to consider them as quantitative methods. Promising results further revealed that the analyzer has a good negative predictive value (NPV) for microbiologically negative cultures, especially for cultures with bacterial counts of 10(5)/l (NPV = 95%). CONCLUSION: The analyzer is capable of providing rapid and reliable urine analysis of cellular particles avoiding the known imprecision of dipstick and sediment methodology. Thus, when used in an algorithm, combined with dipstick or quantitative urine chemistry analysis (for hemoglobin, esterase, protein, glucose, etc.), this analyzer might serve as a rapid and accurate screening tool in routine urine analysis, thereby reducing manual reviewing rate as well as the number of missed samples, compared to screening with dipstick alone. PMID- 11393385 TI - Urinary excretion of vasoactive substances in chronic renal failure. AB - To investigate the pathophysiological role of vasoactive substances in the progression of chronic renal disease, we measured the 24-hour urinary excretion of prostaglandin 6-keto F1alpha, thromboxane B2, NOx, cGMP and ET-1 in 26 patients with chronic renal failure under conservative treatment and in 40 control subjects. Urinary 6-keto PgF1alpha, TxB2 and cyclic GMP were evaluated by RIA, and ET-1 was assayed by EIA. NOx were evaluated using a colorimetric assay as nitrate/nitrite. Urinary excretion of prostaglandin 6-keto F1alpha averaged 18.1 +/- 20.9 ng/g Ucreat in patients vs. 240.9 +/- 257.3 in controls (p < 0.0001), thromboxane B2 422 +/- 374 ng/g Ucreat in patients vs. 967 +/- 589 in controls (p < 2x 10(-5)), NOx 7.07 +/- 5.54 mg/g Ucreat in patients vs. 9.79 +/- 3.77 in controls (p < 0.01), cGMP 310 +/- 200 pg/g Ucreat in patients vs. 488 +/- 241 in controls (p < 0.001). In contrast, ET-1 urinary excretion was almost doubled in patients (13.45 +/- 5.84 ng/g of Ucreat) in comparison with controls (6.84 +/- 2.81 p < 1x10(-5)). While in control subjects significant correlations between urinary excretions of prostaglandin 6-keto F1alpha and thromboxane B2 (r = 0.69, p < 0.001) or NOx and ET-1 (r = 0.54, p < 0.001) were present, in patients only the relationship between urinary excretions of prostaglandin 6-keto F1alpha and thromboxane B2 (r = 0.53, p < 0.01) was retained. Our data suggest that in the normal kidney a balance between prostaglandin I2 and thromboxane A2, or nitric oxide and endothelin-1 is present, which contributes to hemodynamic regulation and protects this organ from ischemic damage. This balance is abolished in CRF, where a large increment of vasopressor agent endothelin is present, which, joined to a prevalent decrease of prostaglandin I2 synthesis, could contribute to the ischemic and fibrogenetic damage of the kidney, leading to progression of renal disease. PMID- 11393386 TI - Retroperitoneal fibrosis and immune-complex glomerulonephritis. AB - A 63-year-old man with rapidly progressive glomerulonephritis of the immune complex type showed typical findings of idiopathic retroperitoneal fibrosis involving the left ureter resulting in hydronephrosis. Treatment with steroid improved both conditions. Our case and previously reported cases showing the same association of conditions support the hypothesis that the association is not fortuitous but reflects a common immunological mechanism. PMID- 11393387 TI - Nephrotic syndrome due to thrombotic microangiopathy (TMA) as the first manifestation of human immunodeficiency virus infection: recovery before antiretroviral therapy without specific treatment against TMA. AB - BACKGROUND: Among the possible renal complications that can develop a human immunodeficiency virus- (HIV) infected patient, thrombotic microangiopathy (TMA) is one of them. This is a type of vascular lesion more common in HIV patients than in normal population, and sometimes it can be the first manifestation of the HIV infection. METHODS: We present a patient with TMA in whom the subsequent investigation to find the cause of TMA revealed HIV infection and Giardia Lamblia in stool. RESULTS: Before antiretroviral therapy was started the patient began to show recovery of the hemolytic anemia, recovery of the nephrotic syndrome and partial remission of the proteinuria, so that he did not receive specific therapy for TMA. CONCLUSIONS: HIV infection should be suspected in patients presenting with TMA, and a HIV test should be routinely performed as part of the initial clinical evaluation of TMA. If the patients have not developed acquired immunodeficiency syndrome, the prognosis of TMA is equal to non-infected ones. PMID- 11393388 TI - Hydroxyethyl starch-induced renal insufficiency after plasma exchange in a patient with polymyositis and liver cirrhosis. AB - Hydroxyethyl starch (HES) is a macromolecular preparation that has been used as a volume expander since 1991. Renal toxicity of high-dose HES is now well recognized but potential renal toxicity of low-dose HES is poorly documented. Acute renal toxicity of cyclosporin A (CyA) may be responsible for osmotic nephrosis-like lesions. We report here the case of a 30-year-old male who developed cirrhosis due to hepatitis B and delta viruses and polymyositis. Polymyositis was treated with CyA, prednisone and plasma exchanges using low-dose HES as the replacement fluid. Renal insufficiency occurred with biopsy-proven osmotic nephrosis-like lesions, considered to be secondary to HES infusions and/or CyA. PMID- 11393389 TI - A case of bronchiolitis obliterans organizing pneumonia with renal dysfunction after renal transplantation. AB - A 35-year-old male renal transplant patient was admitted with 3-week history of cough and myalgia after a 3-month post-transplantation. The patient's posttransplant course was uncomplicated and had no rejection. But on admission, the serum creatinine was 3.0 mg/dl and high-resolution computed tomography (HRCT) showed bilateral patchy densities and peripheral consolidations. Renal biopsy revealed mononuclear cell infiltration and non-specific tubular injury without evidence of rejection. An open lung biopsy was performed which showed multiple fibroblast plugs filling airspaces that were typical pathologic features of bronchiolitis obliterans with organizing pneumonia (BOOP). After prednisone therapy, the patient's pulmonary symptoms and radiographic findings rapidly recovered and serum creatinine also decreased from 3.0 to 2.1 mg/dl. Although the cause of renal dysfunction was not elucidated, we herein report a rare case of BOOP with renal dysfunction after renal transplantation. PMID- 11393390 TI - T cell extranodal lymphoma case involving pleura and pericardium in a renal transplant patient. AB - Application of immunosuppressive treatment for a long period after organ transplantations suppresses immune system in organ receivers and increases the risk of development of neoplastic diseases along with infections. Among the complications developing after transplantation, post-transplant lymphoproliferative diseases are not rare. Although the disease is generally of B cell origin, cases of rare post-transplant lymphoproliferative disease of T cell origin have been reported. Post-transplant lymphoproliferative disease often occurs after Epstein-Barr virus (EBV) infection. In our case an extranodal T cell lymphoma originating from the pleura and pericardium in a renal transplant patient has been diagnosed with cytology. PMID- 11393391 TI - Brown tumor of the thoracic spine in a patient on long-term hemodialysis. AB - A 39-year-old woman on long-term hemodialysis presented with a history of rapidly progressive paraplegia. Radiological examination showed a compression fracture of seventh thoracic vertebra and expansive mass lesion in the posterior elements of the fourth thoracic vertebra. Laboratory tests on admission showed serum calcium of 11.9 mg/dl, phosphate 6.0 mg/dl, and the high-sensitive parathyroid hormone level of 139,191 pg/ml measured by radioimmunoassay. Percutaneous biopsy of the expansive mass showed a large number of multinucleated giant cells in a fibroblastic stroma containing abundant hemosiderin. Tumor resection and anterior interbody fusion with artificial bone graft was performed on 14th hospital day. Paraplegia gradually improved postoperatively. Total parathyroidectomy and autotransplantation of parathyroid gland were subsequently performed. Nodular hyperplasia was evident in the parathyroid glands by light microscopy. Brown tumor is rarely found in vertebral bone and this is the sixth case of such tumor in secondary hyperparathyroidism. PMID- 11393392 TI - Effectiveness of a second course of cyclophosphamide therapy in children with frequently relapsing nephrotic syndrome after cyclosporin A therapy. PMID- 11393394 TI - RPA position on the role of non-physician medical personnel in delivering nephrologic care. Part 3 of 3. PMID- 11393393 TI - Antiribosomal P antibodies in patients with lupus glomerulonephritis. PMID- 11393395 TI - NMR solution structure and conformational analysis of the calcium release agent cyclic adenosine 5'-diphosphate ribose (cADPR). AB - A high-resolution NMR study of the solution structure of the calcium release agent cADPR has been performed. Pseudorotationals analysis reveals that in solution both sugar rings in cADPR adopt predominantly (approximately 75%) South conformations, with the A and N rings adopting approximately 2T3 (C2'-endo(major) C3'-exo(minor) and 4(3)T (C3'-exo-C4'-endo) conformations, respectively. The backbone torsion angles beta and gamma have also been determined. While the minor North conformers were not observed in the crystal structure of cADPR, the solution values of the major South conformers compare well to those found in crystal structure. PMID- 11393396 TI - Photochemical synthesis of novel dideoxynucleosides. AB - A series of 2',3'-dideoxynucleosides based on the apiose family was prepared from photochemical ring-expansion of a common cyclobutanone precursor. The starting ketone, (+/-) 3-[2'-(benzoyloxy)ethyl]-2,2-dimethylcyclobutanone (12) was prepared from commercially available (+/-)alpha-pinene. Since the optically pure antipodes of alpha-pinene are also commercially available, these nucleosides can be prepared optically pure using the identical procedure. PMID- 11393397 TI - An improved synthesis of the dinucleotides pdCpA and pdCpdA. AB - An improved route was developed for the preparation of the dinucleotide hybrid 5' O-phosphoryl-2'-deoxycytidylyl-(3'--> 5')adenosine (pdCpA) 7. This simple and concise synthesis involves the successive coupling of 2-cyanoethyl N, N, N', N' tetra- isopropylphosphorodiamidite with 4-N-benzoyl-5'-O-(4, 4'-dimethoxytrityl) 2'-deoxy-cytidine 1 and 6-N,6-N,2'-O,3'-O-tetrabenzoyladenosine 2 as the key step. Some dinucleotide derivatives bearing different protecting groups were also synthesized and the selective deprotection conditions were studied in detail. The utility and efficiency of this approach has been further demonstrated by its application to the synthesis of total DNA dinucleotide pdCpdA 17 and total RNA dinucleotide 21. PMID- 11393398 TI - A facile synthesis of 5'-end solid-anchored, 3'-end free oligodeoxyribonucleotides via the (5'-->3')-elongated phosphoramidite strategy. AB - It is demonstrated that not only N2- but also O6-protection of the guanine base is necessary for obtaining the oligodeoxyribonucleotides in high yields and at a high purity in the solid-phase synthesis via the (5'--> 3')-chain elongated phosphoramidite approach. PMID- 11393399 TI - Efficient synthesis of fused isothiazolo C-nucleosides. III. Synthesis of substituted isothiazol. AB - The Divakar-Reese procedure has been successfully applied for transforming 7-oxo isothiazolo[4,5-d]pyrimidine C-nucleosides (4a,b, 5a,b, 6a) via 1,2,4-triazol-1 yl intermediates (7a,b, 8a,b) into various 7-substituted C-nucleosides 15a,b, 16a,b, 17a, 18a, 19a,b, 20a,b; their subsequent deprotection provides novel types of unusual C-glycosides 22b, 23a, 24a,b, 25b, 26b. C-Nucleosides, possessing on its heterocyclic base other than naturally occuring oxo- or amino substituents, are important model compounds for biological or medicinal studies (2,3). We want to report on the synthesis of novel 7-substituted isothiazolo = [4,5-d]pyrimidine C-nucleosides. As we could show in previous papers (1,4), there exists a simple approach to the protected C-glycosides 4-6. PMID- 11393400 TI - The production and evaluation of antibodies for enzyme immunoassay of AZTTP. AB - We describe the development of the first enzyme immunoassay for quantifying AZTTP that does not use of radioactive labeling. Anti-AZTTP antibodies were raised in rabbits by immunizing with an AZTTP-kelhoyle limpet hemocyanin (KLH) conjugate. Competitive immunoassays indicated a nanomolar sensitivity to AZTTP. One of the antisera produced was specific for AZTTP. PMID- 11393401 TI - Synthesis and properties of oligonucleotides containing 8-bromo-2' deoxyguanosine. AB - The preparation of oligonucleotides containing 8-bromo-2'-deoxyguanosine is described. Substitution of G by 8-bromoguanine on an alternating CG decamer stabilizes the Z-form in such a way that the B-form was not observed. Melting temperatures showed that duplexes in which 8-bromo-2'-deoxyguanosine paired with natural bases were much less stable. PMID- 11393402 TI - A neural network to predict melting temperature (Tm) of RNA duplex. AB - A straightforward method to predict RNA duplex stability by neural network is described. The best network consists of three layers in which the input layer units are 12 (frequencies of 10 nearest-neighbors and 2 terminals), the hidden layer units are 3 and the output layer unit is 1 (measured Tm). This method to predict Tm has the advantage that the determinations of thermodynamic parameters is not needed. PMID- 11393403 TI - Glomerulonephritis associated with Staphylococcus aureus infection. PMID- 11393404 TI - The natural history of alcoholic chronic pancreatitis. AB - An improved knowledge of the natural history is the indispensible basis for a rational concept in regard to the diagnosis, classification, understanding and management of pain in chronic pancreatitis. Unfortunately, data on the natural history of CP are scarce and conflicting. Some relevant observations of our prospective long-term study of a mixed medical-surgical cohort comprising 207 patients with alcoholic CP (mean follow-up 17 years from onset) are summarized. In early-stage CP, episodes of recurrent pancreatitis were predominant. Severe persistent pain was typically associated with local complications (mainly postnecrotic cysts in 54%; symptomatic cholestasis in 24%) relieved definitely by a drainage procedure. Lasting pain remission was documented in >80% of the whole cohort within 10 years from onset in association with marked pancreatic dysfunction. From our experience, the relief of "chronic" pain regularly follows selective surgery tailored to the presumptive pain cause or it occurs spontaneously in uncomplicated advanced CP (excluding narcotic addiction). PMID- 11393405 TI - Semiquantitative measurement of acetylcholine receptor at the motor end-plate in myasthenia gravis. AB - OBJECTIVE: The purpose of this study was to investigate whether this semiquantitative measurement of the motor end-plate acetylcholine receptors (AChRs) can be used to confirm the diagnosis of myasthenia gravis (MG), and in particular ocular MG. METHODS: Motor point biopsies were performed from the biceps brachii muscles. Measurement of AChRs was made in peroxidase-labeled alpha bungarotoxin stained muscle specimens. PATIENTS: Twenty patients with ocular MG, 37 with generalized MG, 5 with Lambert-Eaton myasthenic syndrome, 3 with botulism, 8 with amyotrophic lateral sclerosis, and 8 controls were included in this study. RESULTS: AChRs were decreased in all patients with generalized MG and in 80% of ocular MG including patients without detectable circulating anti-AChR antibodies, as compared with the control subjects. CONCLUSION: This method is useful to confirm the diagnosis of MG, in particular ocular MG without detectable anti-AChR antibodies. PMID- 11393406 TI - Cholangiocarcinoma with a background of hepatitis B virus-associated cirrhosis. AB - Recently, hepatitis virus-associated chronic hepatitis or cirrhosis has been suggested to be involved in the pathogenesis of cholangiocarcinoma (CC). A 52 year-old man was diagnosed as CC with a background of hepatitis B virus (HBV) dependent cirrhosis. A minute hepatic tumor was found during the follow-up, and was diagnosed as CC on percutaneous biopsy. The patient died of hepatic failure and an autopsy revealed the tumor to be a well to moderately differentiated adenocarcinoma. An immunohistological analysis of HBV X gene-encoded protein (HBX) was neither detected in the cancerous nor in the noncancerous tissue. No oncogenic role of the virus was verified in this case. PMID- 11393407 TI - Insulinoma with subsequent association of Zollinger-Ellison syndrome. AB - We report a patient with insulinoma associated with Zollinger-Ellison syndrome. A 67-year-old woman was first admitted to our hospital for an abdominal mass. Abdominal computed tomography (CT) revealed a large pancreatic tumor, which was then diagnosed as an unresectable pancreatic adenocarcinoma. At the age of 71, she presented symptoms of hypoglycemia. Fasting blood glucose was 21 mg/dl and plasma immunoreactive insulin level was 846 microU/ ml. Plasma gastrin, glucagon, vasoactive intestinal polypeptide and somatostatin levels were all normal. At the age of 73, hypoglycemic attacks occurred more frequently and she was admitted to our hospital. Abdominal CT scan showed multiple liver metastases. Chemotherapy with 5-fluorouracil and doxorubicin was performed. Three months later, she had an emergency laparotomy because of a perforated duodenal ulcer. Plasma gastrin level was 1,960 pg/ml at that time. Gastric hypersecretion was well controlled with a proton pump inhibitor (lansoprazole) but she died of widespread cancer dissemination 8 years after her first admission. On autopsy, histologic examination revealed a mixed acinar-endocrine carcinoma of the pancreas. Immunohistochemical stains were positive for insulin, gastrin, and alpha1 antitrypsin. PMID- 11393408 TI - Primary cardiac angiosarcoma detected by magnetic resonance imaging but not by computed tomography. AB - A 51-year-old man with a primary angiosarcoma of the right atrium is reported. The angiosarcoma was not detected by transthoracic echocardiography or computed tomography, but magnetic resonance imaging and transesophageal echocardiography did show the tumor of the right atrial free wall. We performed a transvenous endomyocardial biopsy of the tumor under the guidance of transesophageal echocardiography and made the pathological diagnosis. This case demonstrates the advantage of magnetic resonance imaging and transesophageal echocardiography for tumor detection over transthoracic echocardiography and computed tomography and the usefulness of transesophageal echocardiography for guiding the right atrial endomyocardial biopsy procedure. PMID- 11393409 TI - Serious arrhythmias in patients with apical hypertrophic cardiomyopathy. AB - We report cases of serious arrhythmias associated with apical hypertrophic cardiomyopathy (AHCM). Thirty-one patients were referred to our institute to undergo further assessment of their AHCM from 1988 to 1999. Three patients with nonsustained ventricular tachycardia demonstrated an 123I-MIBG regional reduction in the tracer uptake. In two patients with ventricular fibrillation (VF), the findings from 123I-MIBG imaging revealed regional sympathetic denervation in the inferior and lateral regions. Electrophysiologic study demonstrated reproducible induction of VF in aborted sudden death and presyncopal patients, resulting in the need for an implantable defibrillator device and amiodarone in each patient. Patients with refractory atrial fibrillation with a rapid ventricular response suffered from serious congestive heart failure. A prudent assessment and strategy in patients with this disease would be indispensable in avoiding a disastrous outcome. PMID- 11393410 TI - Isolated tricuspid valve endocarditis due to Candida parapsilosis associated with long-term central venous catheter implantation. AB - A 72-year-old man was treated for fungal tricuspid valve endocarditis (TVE) with significant tricuspid valvular regurgitation and severe congestive heart failure caused by Candida parapsilosis. The patient had received hyperalimentation and antibiotic therapy for three months through a central venous catheter after the surgical treatment of ileus. The patient was treated medically with amphotericin B and fluconazole because of high surgical risk due to severe pulmonary emphysema, and he responded well. Although TVE caused by C. parapsilosis is rare, we should consider this possibility in patients receiving long-term hyperalimentation and antibiotic therapy using a central venous catheter. PMID- 11393411 TI - An 80-year-old mitochondrial disease patient with A3243G tRNA(Leu(UUR)) gene presenting cardiac dysfunction as the main symptom. AB - MELAS is characterized by mitochondrial myopathy, encephalopathy, lactic acidosis and stroke-like episodes, but cardiac involvement also frequently occurs. An 80 year-old female patient had been suffering from insulin-dependent diabetes mellitus and neurosensory hearing loss. At the age of 79 she suffered metabolic acidosis with persistent drowsiness and was subsequently found to have severe cardiac dysfunction. Muscle biopsy disclosed the presence of abnormal mitochondria, and the MELAS gene mutation (A3243G of the tRNA(Leu(UUR))) was demonstrated. It is noteworthy that this mitochondrial disease patient has survived until a great age, which shows the wide clinical spectrum of MELAS, especially in the age of onset. PMID- 11393412 TI - Adult T-cell leukemia with hypercalcemia-induced metastatic calcification in the lungs due to production of parathyroid hormone-related protein. AB - A 60-year-old man was diagnosed as adult T-cell leukemia with severe hypercalcemia because of production of parathyroid hormone-related protein. After admission, the patient had respiratory insufficiency with an infiltrative shadow in his lungs suggestive of pneumonia. However, neither improvement in respiratory function nor disappearance of the abnormal chest shadow was observed with administration of various antibiotics. An autopsy demonstrated the chest shadow had been caused by metastatic calcification associated with hypercalcemia due to production of parathyroid hormone-related protein. The possibility of metastatic calcification should be considered in patients with adult T-cell leukemia and hypercalcemia who have an abnormal chest shadow. PMID- 11393413 TI - Hypopituitarism due to pituitary metastasis of lung cancer: case of a 21-year-old man. AB - A 21-year-old man presented persistent dry cough, general malaise, loss of appetite, decreased sexual desire and double vision. Chest radiograph revealed a mass shadow in the left upper lobe. Histopathological diagnosis of the tumor was squamous cell carcinoma. Brain computed tomography and magnetic resonance imaging revealed a metastasis to the pituitary gland. Hypopituitarism was diagnosed by pituitary function tests. Diabetes insipidus was absent and the function of the posterior lobe of the pituitary gland was preserved. Hypopituitarism due to pituitary metastasis is a rare complication of lung cancer, and has never been reported in a patient as young as 21 years old. PMID- 11393414 TI - Diabetic ketoacidosis associated with recurrent pulmonary edema and rhabdomyolysis in a patient with Turner's syndrome. AB - Turner's syndrome is a condition involving total or partial absence of one X chromosome and has been associated with a number of diseases including non insulin dependent diabetes mellitus, abnormalities of glucose metabolism and hypothreosis. There have been many case reports in which Turner's syndrome is associated with type 2 diabetes, but the association with type 1 diabetes and/or life threatening complications is very rare. We present an unusual case of a patient with Turner's syndrome who has type 1 diabetes and is complicated with ketoacidosis, severe acute and recurrent pulmonary edema and rhabdomyolysis. PMID- 11393415 TI - Visual disturbance due to carotid artery thrombosis in a patient with familial hypercholesterolemia; response to surgical thrombotectomy. AB - A 48 years-old Japanese man suffered from marked xanthomas on ankles, knees, hand fingers, and foot joints due to insufficient control of serum hypercholesterolemia despite low density lipoprotein (LDL-C) absorptive therapy followed by treatment with potent anti-hypercholesterolemic agents. He had undergone surgical resection of xanthoma on the knee, foot and hand finger joints. Treatment with simvastatin returned the serum total cholesterol levels to nearly normal levels, followed by marked fluctuations. He subsequently experienced transient right-visual disturbance, and roentogenographic examination was performed. The patient was diagnosed as right-common carotid artery thrombosis. After the thrombotectomy of the right-common carotid artery, his visual power was markedly improved. PMID- 11393417 TI - Spontaneous rupture of the amyloid spleen in a case of usual interstitial pneumonia. AB - We describe a rare case of secondary amyloidosis associated with usual interstitial pneumonia (UIP), who died from spontaneous rupture of the amyloid spleen. A 68-year-old man was admitted to evaluate to his interstital lung disease. Chest radiography showed reticular shadow in bilateral lower lung fields. Two years later, he suddenly felt severe abdominal pain. In spite of maximum therapy, he died from hypovolemic shock. Postmortem examination revealed massive intraabdominal hemorrhage. The diagnosis of lung disease was UIP and amyloid A type deposits were observed in various organs including the ruptured spleen. PMID- 11393416 TI - Glomerulonephritis after methicillin-resistant Staphylococcus aureus infection resulting in end-stage renal failure. AB - A 58-year-old man developed proteinuria and renal dysfunction following pneumonia caused by methicillin-resistant Staphylococcus aureus (MRSA). Vancomycin was administered, and prednisolone pulse therapy and plasmapheresis were performed. Subsequently, serum creatinine was decreased. Eight months later, creatinine and CRP were again elevated, and MRSA was detected. Vancomycin was again administered and plasmapheresis was performed. However, renal function was not improved and continuous hemodialysis was initiated. This case indicates that complete eradication of MRSA is necessary to treat MRSA-associated glomerulonephritis, and if this is not attained, a permanent loss of renal function occurs. PMID- 11393418 TI - Churg-Strauss syndrome after reduction of inhaled corticosteroid in a patient treated with pranlukast for asthma. AB - Recently, various forms of Churg-Strauss syndrome (CSS) have been reported in association with the use of leukotriene receptor antagonists. A 53-year-old woman with a 5-year history of asthma associated with chronic sinusitis presented mononeuropathy, hypereosinophilia, and positive P-ANCA in October 1999. She had been treated with pranlukast (450 mg/day) and beclomethasone dipropionate (BDP) at a dose of 1,200 microg/day which had gradually been tapered to 800 microg/day over the previous 17 months. She was found to have CSS, and 60 mg/day of prednisolone was administered instead of pranlukast, resulting in an improvement of her symptoms and eosinophilia. Later, we confirmed that serum P-ANCA had been positive before the pranlukast treatment, but CSS vasculitis had not appeared at that time. We speculated that an underlying incomplete form of CSS was being masked in this case and that the reduction of inhaled corticosteroid might have been responsible for the unmasking of CSS. PMID- 11393419 TI - Malignant lymphoma of the bone associated with systemic sarcoidosis. AB - A 57-year-old woman was hospitalized with malignant lymphoma of the right talus. After treatment, complete remission was obtained. Gallium-67 scintigraphy to confirm the remission demonstrated increased uptake in the whole body skeletal muscle, especially in her thighs. Biopsy of right gastrocnemius muscle showed epithelioid granuloma. Serum angiotensin-converting enzyme activity (ACE) and lysozyme had increased to several times the normal range. We diagnosed her disease as bone-associated sarcoidosis-lymphoma syndrome. Human herpes virus 8 (HHV-8) genome was examined in the bone marrow specimen, and the relationship between sarcoidosis-lymphoma syndrome and HHV-8 was discussed. PMID- 11393420 TI - Neurosyphilis showing transient global amnesia-like attacks and magnetic resonance imaging abnormalities mainly in the limbic system. AB - We report a case of neurosyphilis with transient global amnesia (TGA)-like attacks on the first presentation. MRI abnormalities in bilateral limbic systems, including a few lesions in the basal ganglia and thalamus, were identified. Depression and dementia became apparent, accompanied by a high treponemal antibody titer and mild cortical atrophy. Antisyphilitic therapy brought about mild improvement, and the MRI abnormalities decreased. PMID- 11393421 TI - Bilateral paramedian thalamo-midbrain infarction showing electroencephalographic alpha activity. AB - A 57-year-old man became unresponsive and mute with bilateral ophthalmoplegia and quadriplegia. Magnetic resonance imaging (MRI) showed bilateral infarctions at the ventral midbrain and the dorsomedial nucleus of the thalamus. Serial studies with MR and vertebral angiography disclosed hypoperfusion and spontaneous reperfusion of the bilateral posterior cerebral arteries at their origin from the basilar artery. Electroencephalographically, a posteriorly distributed alpha rhythm was clearly recorded and it was reactive in response to external stimuli. The findings seen in the present patient suggest that the ventral midbrain and medial dorsal thalamus are not necessary to produce posterior electroencephalographic alpha activity. PMID- 11393422 TI - Protein-losing enteropathy exacerbated with the appearance of symptoms of systemic lupus erythematosus. AB - A 38-year-old woman visited our hospital with edema on her face and conjunctivae. The underlying disease was not clarified, and she did not visit the hospital afterwards. She suffered from diarrhea, polyarthralgia, Raynaud's phenomenon, malar rash and hair loss in the subsequent two years, and was hospitalized because of hypoproteinemia. Her urine, liver and heart test results did not account for her hypoproteinemia. She was diagnosed as having protein-losing enteropathy (PLE) associated with SLE based on the 99mtechnetium-labeled human serum albumin scintigraphy findings, clinical findings and laboratory results of antinuclear and anti-Sm antibodies. This case report demonstrates a strong association between PLE and SLE because PLE was aggravated along with the appearance of SLE symptoms and PLE subsided with prednisolone treatment along with improvement of SLE. PMID- 11393423 TI - Two cases of long lasting bacteremia due to Mycobacterium avium complex despite new macrolides-containing regimens in patients with acquired immunodeficiency syndrome. AB - The prognosis of Mycobacterium avium complex (MAC) infection has been improved by new macrolides-containing regimens and the use of highly active antiretroviral therapy (HAART) in the treatment of acquired immunodeficiency syndrome (AIDS). We report on two AIDS cases with long lasting bacteremia due to MAC under this regimen. Both patients experienced problems due to side effects from the anti-MAC regimen and from an immune-reconstitution syndrome related to HAART. MAC infection persisted despite treatment, however, no anti-MAC drug-resistant isolates emerged throughout the clinical course in either case. These cases demonstrate that therapy for disseminated MAC infection is sometimes difficult even with HAART and macrolides-containing regimens. PMID- 11393424 TI - Severe hyponatremia in a patient treated with levomepromazine and carbamazepine. PMID- 11393425 TI - Using insurance to enhance nitrogen fertilizer application to reduce nitrogen losses to the environment. AB - The advantage of using insurance to help a farmer adopt a best nitrogen management plan (BNMP) that reduces the impact of agricultural production on the environment is analytically and empirically demonstrated. Using an expected value analysis, it is shown that an insurance program can be structured so as to reduce a farmer's cost of bearing the adoption risk associated with changing production practices and, thus, to improve the farmer's certainty equivalent net return thereby promoting the adoption of a BNMP. Using the adoption of growing-season only N fertilizer application in Iowa as a case study, it is illustrated how insurance may be used to promote the adoption of this practice to reduce N fertilizer use. It is shown that it is possible for a farmer and an insurance company both to have an incentive to develop an insurance adoption program that will benefit both the farmer and the insurance company, increasing net social welfare and improving environmental quality in Iowa. PMID- 11393426 TI - Adsorption behaviour of some aromatic amines on pyrolusite and activated carbon and recovery of beta napthylamine from water sample. AB - The adsorption behaviour of Diphenylamine (DPAM), beta napthylamine (beta NAM), alpha napthylamine (alpha NAM) and aniline on pyrolusite and activated carbon has been studied. Pyrolusite shows remarkable sorption capacity for DPAM and beta NAM as compared to aniline; (the adsorption followed the order: Activated Carbon:--> DPAM = beta NAM > alpha Aniline; Pyrolusite: DPAM: --> beta NAM > alpha NAM > Aniline) The maximum adsorption of beta NAM occurred in the concentration range 4 20 microg mL(-1) on pyrolusite (95%) and 4-50 microg mL(-1) on activated carbon (100%). The effect of various doses of activated carbon on the adsorption of beta NAM confirm Langmuir and Freundlich isotherms where as Freundlich isotherm is obeyed by pyrolusite. The adsorption of beta NAM on both the absorbents is not affected in presence of DPAM over a wide range of their initial concentrations (20-60 microg mL(-1)). The desorption studies of beta NAM on pyrolusite was carried out by batch as well as column processes. Excellent results were obtained when a mixture of n-hexane and isopropanol (91:1) was used as eluent. PMID- 11393427 TI - A multivariate analysis of biophysical parameters of tallgrass prairie among land management practices and years. AB - Six treatments of eastern Kansas tallgrass prairie--native prairie, hayed, mowed, grazed, burned and untreated--were studied to examine the biophysical effects of land management practices on grasslands. On each treatment, measurements of plant biomass, leaf area index, plant cover, leaf moisture and soil moisture were collected. In addition, measurements were taken of the Normalized Difference Vegetation Index (NDVI), which is derived from spectral reflectance measurements. Measurements were taken in mid-June, mid-July and late summer of 1990 and 1991. Multivariate analysis of variance was used to determine whether there were differences in the set of variables among treatments and years. Follow-up tests included univariate t-tests to determine which variables were contributing to any significant difference. Results showed a significant difference (p < 0.0005) among treatments in the composite of parameters during each of the months sampled. In most treatment types, there was a significant difference between years within each month. The univariate tests showed, however, that only some variables, primarily soil moisture, were contributing to this difference. We conclude that biomass and % plant cover show the best potential to serve as longterm indicators of grassland condition as they generally were sensitive to effects of different land management practices but not to yearly change in weather conditions. NDVI was insensitive to precipitation differences between years in July for most treatments, but was not in the native prairie. Choice of sampling time is important for these parameters to serve effectively as indicators. PMID- 11393429 TI - Marine eutrophication: a proposed data analysis procedure for assessing spatial trends. AB - A methodology for the discrimination of the different trophic levels at a spatial scale in the marine environment is proposed using spatial analysis methods and non-parametric statistics. Phytoplankton cell number, being a representative parameter to express trophic trends in the marine ecosystem is selected for the methodology development; Saronicos Gulf, Greece is used for the case study. The proposed stepwise methodology includes interpolation for assessing the spatial distribution of phytoplankton cell number, division of the Gulf into a number of quadrates, development of a scale characterising trophic levels and finally characterisation of the trophic state of each quadrate using non-parametric statistics. The advantages of this methodology and the potential applications in coastal management studies are also discussed. PMID- 11393428 TI - Exploratory analysis and a stochastic model for humusdisintegration. AB - Ulrich (1981) supposes in the hypothesis of humusdisintegration that the balance between polymerisation and breakdown of organic material may be disturbed in chemically well buffered European forest soils. This new aspect of aluminium toxicity may cause nitrogen exceedance in forest ecosystems and subsequent seasonal nitrate output (Eichhorn and Hutterman, 1999). In a research program the substances in the seepage water are monitored in a small woodland in central Germany. We explore these multivariate data for examining possible influences on the process of humusdisintegration and its temporal evolution. As a result, a regression model for carbon is developed, which includes covariables, i.e., other substances, as well as spatial and temporal terms describing systematic variability. Especially iron and aluminium turn out to be very influential in the model. So far our work is a basic step for monitoring the seepage water data by means of stochastic modelling. PMID- 11393430 TI - Environmental monitoring in the The Netherlands: past developments and future challenges. AB - During the past three decades environmental monitoring systems covering an immense variety of environmental problems emerged rapidly throughout the world. In The Netherlands the entire spectrum of monitoring systems is present. Their development is especially interesting because of the Dutch complex environmental landscape: a result of the combination of a natural setting with a high small scale variation and an intense use of the available space by both industry and agriculture. This necessitates a well balanced environmental policy wherein monitoring of both environmental compartments and environmental policy itself plays an important role. In this article we focus on abiotic systems, whereby the development of monitoring will be related to that of environmental policy in general. Some general lines emerge: from quantity towards quality, from single pollutants towards environmentally harmful processes as a whole, and from a focus on human health and safety towards integrated environmental management. Because The Netherlands is a highly organized and highly educated society, reacting fast and flexible to new problems, this progression has happened quickly. However, still much remains to be desired and flexibility not necessarily guarantees optimal routes of development. Paramount is the development of sophisticated and robust monitoring systems that truly support integrated environmental management. This asks for clear objectives based on solid scientific insights. PMID- 11393431 TI - Is all sexual harassment viewed the same? Mock juror decisions in same- and cross gender cases. AB - Given recent court decisions, there is a need to investigate less common forms of sexual harassment, including women harassing men and same-gender harassment. The present study was a 2 (harasser gender) x 2 (target gender) x 2 (participant gender) factorial design in which 408 mock jurors made decisions in a hostile work environment case. Women harassing men were more likely to be found guilty than were men harassing women, and harassers in same-gender cases were more likely to be found guilty and were perceived more negatively than harassers in cross-gender cases. Participant gender differences were found in cross-gender, but not same-gender, conditions. Results suggest that the gender composition of the harasser and target may be an extralegal factor influencing managerial and juror decision making. PMID- 11393432 TI - Assuming preferential selection when the admissions policy is unknown: the effects of gender rarity. AB - One hundred thirty-five undergraduates indicated the degree to which they believed gender played a role in the selection of an applicant for a graduate degree program. Both the gender composition of the cohort and the selection policy (explicitly merit-based, explicitly affirmative action, or ambiguous) were varied. Results indicated that preferential selection on the basis of gender was assumed when women were solos and explicit information about the selection policy was not provided and that these assumptions were as strong as when an affirmative action policy was explicitly stated. This did not occur when the female selectee was not a solo or when a male selectee was a solo. Evaluations of qualifications and prediction of success paralleled the preferential selection assumptions. PMID- 11393433 TI - Organizational attractiveness of firms in the People's Republic of China: a person-organization fit perspective. AB - The authors investigated factors related to firm attractiveness as an employer in the People's Republic of China. The organizational attributes of type of ownership, nationality of the supervisor, and firm familiarity in organizational descriptions were manipulated and their effects were measured on firm attractiveness. In addition, the authors adopted a person-organization fit perspective to investigate how individual difference characteristics moderated the effects of these organizational attributes on attractiveness. Although, in general, participants were more attracted to foreign than state-owned firms and to familiar than unfamiliar firms, results provided support for the person organization fit perspective in that the individual differences moderated the effects of the organizational attributes on firm attractiveness. For example, participants were more attracted to state-owned versus foreign firms when they were more risk averse and had a lower need for pay. Thus, the results provide initial support for the generalizability of the person-organization fit perspective to a non-Western setting. PMID- 11393434 TI - Aging and lineup performance at long retention intervals: effects of metamemory and context reinstatement. AB - Young (18-30 years) and older (62-79 years) adults (N = 96) engaged in a 20-min live interaction with the future target in a lineup task. One month later, participants were interviewed about the events in the prior encounter (with or without context reinstatement), and then they saw a target-present (TP) or target absent (TA) lineup. The lineup was followed by the Benton Face Recognition Test (A. Benton, A. Sivan, K. Hamsher, N. Varney, & O. Spreen, 1994), which correlated positively with accuracy in TP, especially for young adults. False identification in TA was associated with (a) higher scores on a memory self-efficacy scale and (b) higher recall of information about the initial event, although only for seniors. Results suggested that age-related increases in false identification generalize to ecologically valid conditions and that seniors' performance on lineups is negatively related to verbal recall as well as to self-reports of satisfactory experiences with memory in life. PMID- 11393435 TI - Are performance appraisal ratings from different rating sources comparable? AB - The purpose of this study was to test whether a multisource performance appraisal instrument exhibited measurement invariance across different groups of raters. Multiple-groups confirmatory factor analysis as well as item response theory (IRT) techniques were used to test for invariance of the rating instrument across self, peer, supervisor, and subordinate raters. The results of the confirmatory factor analysis indicated that the rating instrument was invariant across these rater groups. The IRT analysis yielded some evidence of differential item and test functioning, but it was limited to the effects of just 3 items and was trivial in magnitude. Taken together, the results suggest that the rating instrument could be regarded as invariant across the rater groups, thus supporting the practice of directly comparing their ratings. Implications for research and practice are discussed, as well as for understanding the meaning of between-source rating discrepancies. PMID- 11393436 TI - The effects of defendant conduct on jury damage awards. AB - Laws of negligence dictate that jurors' decisions about damages be influenced by the severity of plaintiffs' injuries and not by the reprehensibility of defendants' conduct. The authors simulated an automobile negligence trial to assess whether jurors' decisions are in accord with those expectations. Conduct of the defendant and severity of the plaintiff's injuries were manipulated. Jurors listened to the evidence, completed predeliberation questionnaires, deliberated as a jury, and completed postdeliberation questionnaires. Severity of the plaintiff's injury had a strong impact on damage awards, but evidence related to the defendant's conduct was also influential, particularly when the plaintiffs injuries were mild. Here, jurors with any conduct-related evidence gave larger damage awards than jurors with no conduct-related evidence. Findings suggest an effect of defendant conduct on damage awards that may be mediated by judgments that the defendant was negligent. PMID- 11393437 TI - Forgotten partners of international assignments: development and test of a model of spouse adjustment. AB - The authors developed and tested a model of spouse adjustment to international assignments in a sequence of qualitative and quantitative investigations. From in depth interviews with expatriate spouses, half of whom had either positive or negative experiences living abroad, the authors identified several potentially important sources of adjustment. These constructs were integrated with others from identity theory and earlier research on expatriates to develop a conceptual model of spouse adjustment. The model was tested by collecting multisource questionnaire data from 221 international assignee couples working in 37 countries. Correlation and regression analyses revealed several important relationships between individual, interpersonal relationship, and environmental sources of identity, and the adjustment of expatriate spouses. PMID- 11393438 TI - Assessor training strategies and their effects on accuracy, interrater reliability, and discriminant validity. AB - This study compares the effects of data-driven assessor training with schema driven assessor training and control training. The sample consisted of 229 industrial and organizational psychology students and 161 managers who were randomly assigned to 1 of these training strategies. Participants observed and rated candidates in an assessment center exercise. The data-driven and schema driven assessor training approaches outperformed the control training on all 3 dependent variables. The schema-driven assessor training resulted in the largest values of interrater reliability, dimension differentiation, and accuracy. Managers provided significantly more accurate ratings than students but distinguished less between the dimensions. Practical implications regarding the design of assessor trainings and the composition of assessor teams are proposed. PMID- 11393439 TI - Individual differences in utilizing control to cope with job demands: effects on susceptibility to infectious disease. AB - This study examined the interactive effects of job demands, control, and individual characteristics on upper respiratory illnesses and immune function. Having high job control appeared to lessen the linkage between job demands and poor health among individuals with high self-efficacy and those who perceived that they were not often responsible for negative job outcomes. Conversely, having high job control exacerbated the association between job demands and poor health among inefficacious individuals. Implications for promoting more healthful work environments and facilitating employee coping are discussed. PMID- 11393440 TI - To transfer or not to transfer? Investigating the combined effects of trainee characteristics, team leader support, and team climate. AB - Eighty pilots participated in a study of variables influencing the transfer process. Posttraining performance was assessed in a flight simulation under 1 of 2 conditions. Those in the maximum performance condition were made aware of the skill to be assessed and the fact that their teammates were confederates, whereas those in the typical performance condition were not. The results indicated that (a) simulator ratings correlated with a measure of transfer to the cockpit for those in the typical condition only; (b) team leader support, manipulated in a pretask brief, moderated the disparity between maximum and typical performance; (c) team climate mediated the impact of support on performance in the typical condition; (d) those with a stronger predisposition toward the trained skill viewed their climate as more supportive; and (e) perceptions of team climate were better predictors of performance for those with a more external locus of control. PMID- 11393441 TI - Labeling sexual harassment in the military: an extension and replication. AB - Research conducted by V. Magley, C. Hulin, L. F. Fitzgerald, and M. DeNardo (1999) has suggested that women who experience sexual harassment report worse outcomes independent of the labeling process. This study replicates and extends that work. Discriminant analyses were conducted on a sample of approximately 28,000 men and women from the military. The authors included variables similar to those used by V. Magley et al., as well as a variety of antecedent variables. Two significant functions were obtained from the discriminant analysis. The 1st function ordered groups according to the frequency of harassment and accounted for substantially more variance than did the 2nd function, which ordered groups according to whether they labeled their experiences as sexual harassment. The overall results from these analyses demonstrate that labeling incidents as sexual harassment is of marginal meaningfulness in terms of job outcomes and antecedents of harassment. PMID- 11393442 TI - Personnel selection and the five-factor model: reexamining the effects of appplicant's frame of reference. AB - Recently, 2 separate yet related criticisms have been levied against the adequacy of the five-factor model (or Big Five) as a descriptive taxonomy of job applicant personality: frame of reference effects (M. J. Schmit & A. M. Ryan, 1993) and socially desirable responding (A. F. Snell & M. A. McDaniel, 1998). Of interest, although both criticisms suggest that the five-factor model is inadequate, the frame of reference effects criticism suggests that the factor structure should be more complex, whereas socially desirable responding suggests that it should be less complex in job applicant contexts. The current research reports the results of a new study demonstrating the adequacy of the five-factor model as a descriptor of job applicant, job incumbent, and student personality. Implications for personality assessment and concurrent validation designs using personality measures are also discussed. PMID- 11393443 TI - The role of justice in team member satisfaction with the leader and attachment to the team. AB - This study examined the effects of team decision accuracy, team member decision influence, leader consideration behaviors, and justice perceptions on staff members' satisfaction with the leader and attachment to the team in hierarchical decision-making teams. The authors proposed that staff members' justice perceptions would mediate the relationship between (a) team decision accuracy, (b) the amount of influence a staff member has in the team leader's decision, and (c) the leader's consideration behaviors and staff attachment to the team and satisfaction with the leader. The results of an experiment involving 128 participants in a total of 64 teams, who made recommendations to a confederate acting as the team leader, generally support the proposed model. PMID- 11393444 TI - Voice and cooperative behavior as contrasting forms of contextual performance: evidence of differential relationships with big five personality characteristics and cognitive ability. AB - The results of a laboratory study of 276 individuals replicate past findings for cooperative behavior as a form of contextual performance and extend past research by providing evidence that voice (constructive change-oriented communication) may be another form of contextual performance. Conscientiousness, extraversion, and agreeableness related more strongly to voice behavior and cooperative behavior than to task performance. Cognitive ability related more strongly to task performance than to voice behavior or cooperative behavior. Results also demonstrate contrasting relationships for agreeableness (positive with cooperative behavior and negative with voice behavior). This supports recent research suggesting the possibility of bidirectional relationships with personality characteristics across different dimensions of job performance. PMID- 11393445 TI - Prospective relationships between drug problems and work adjustment in a community sample of adults. AB - The prospective relationships between drug problems and work adjustment (e.g., job instability, job satisfaction) were examined in a community sample of 470 adults. Polydrug problems (alcohol, marijuana, and cocaine problem drug use) were both predictors and consequences of work adjustment. In partial support of the impaired functioning theory, polydrug problems predicted reduced job satisfaction 4 years later. Supporting the work-related strain theory, early job instability predicted polydrug problems 4 years later. In support of the theory of general deviance, low social conformity predicted later job instability. Finally, supporting social support theory, early support for drug problems reduced polydrug problems and increased job satisfaction 4 years later. PMID- 11393446 TI - Achieving desired images while avoiding undesired images: exploring the role of self-monitoring in impression management. AB - A study was conducted to test the hypothesis that high self-monitors more effectively manage impressions than low self-monitors do. Students in work groups indicated the extent to which they used 5 impression-management tactics over the course of a semester-long project. At the project's conclusion, students provided their perceptions of the other members of their group. The relationship between impression management and image favorability was then examined across 339 student student dyads. The results generally suggest that high self-monitors can use impression-management tactics more effectively than can low self-monitors. In particular, high self-monitors appear to be more adept than low self-monitors at using ingratiation, self-promotion, and exemplification to achieve favorable images among their colleagues. PMID- 11393447 TI - Method of linearizing the 3d L3/L2 white line ratio as a function of magnetic moment. AB - We have developed a parameterization method which linearizes the relationship between local magnetic moment and the 3d L3/L2 "white line" ratio as observed in electron energy loss spectroscopy or X-ray near edge absorption spectroscopy. We first establish that the parameterization linearizes an existing theoretical result for ratio versus moment. We then test our method on data sets for which a white line ratio has been previously published by other authors, who have studied a series of compounds using a consistent deconvolution procedure. Finally, we apply our linearization method to the observed ratios of a series of 3d transition metals, and to the Cr L edges for a Au(x)Cr(1 - x) alloy. In addition we obtain, for the first time, experimental results on the Au L3 and L2 edge white lines of this alloy system. These results are consistent with a model in which the large local moment in this system is not limited to Cr dopants, but extends into the gold matrix. PMID- 11393448 TI - Quantitative high-resolution microscopy on a suspended chain of gold atoms. AB - High-resolution electron microscope images of a suspended gold chain were analyzed quantitatively. The images of the suspended gold chain were recorded on videotapes, during in situ observation in an ultra-high-vacuum electron microscope. The chain consisted of four gold atoms free from any substrate. Image simulation verified that the gold chain has a center-top geometry. The simulation also showed that the chain has no glue atoms spinning around the axis of the chain. A spinning gold atom was shown to give 50% of the stationary atom in gray scale. By comparing the noise level of the observed image and the gray scale of single atoms such as C, Si, S, Cu, and Au, we verified that the Si and S atoms have almost twice as large contrast as the noise (peak-to-peak), and C atoms have almost the same contrast as the noise level. The analysis showed it critical to detect a single Si and S atoms. PMID- 11393449 TI - High-angle annular dark-field STEM observation of Xe nanocrystals embedded in Al. AB - High-angle annular dark-field scanning transmission electron microscope (HAADF STEM) observation of Xe precipitates embedded in crystalline membranes has been made using electron probes of atomic dimensions and HAADF-STEM images of Xe precipitates qualitatively different from conventional TEM observation results have been obtained. Multislice-based HAADF-STEM simulation has been made and it has been revealed that the intensity of images of Xe atoms at positions displaced from Al matrix columns decreases rapidly as the thickness increases. Even in a thin specimen, the off-site Xe atoms of the precipitate at deep locations, were not observable. Therefore, different images are expected for specimens of different thicknesses or depths of these precipitates. These results indicate that the observation of precipitates in crystalline membranes requires some care. PMID- 11393450 TI - Optimization of scanning transmission X-ray microscopy for the identification and quantitation of reinforcing particles in polyurethanes. AB - The morphology, size distributions, spatial distributions, and quantitative chemical compositions of co-polymer polyol-reinforcing particles in a polyurethane have been investigated with scanning transmission X-ray microscopy (STXM). A detailed discussion of microscope operating procedures is presented and ways to avoid potential artifacts are discussed. Images at selected photon energies in the C 1s, N 1s and O 1s regions allow unambiguous identification of styrene-acrylonitrile-based (SAN) copolymer and polyisocyanate polyaddition product-based (PIPA) reinforcing particles down to particle sizes at the limit of the spatial resolution (50 nm). Quantitative analysis of the chemical composition of individual reinforcing particles is achieved by fitting C 1s spectra to linear combinations of reference spectra. Regression analyses of sequences of images recorded through the chemically sensitive ranges of the C 1s, N 1s and O 1s spectra are used to generate quantitative compositional maps, which provide a fast and effective means of investigating compositional distributions over a large number of reinforcing particles. The size distribution of all particles determined by STXM is shown to be similar to that determined by TEM. The size distributions of each type of reinforcing particle, which differ considerably, were obtained by analysis of STXM images at chemically selective energies. PMID- 11393451 TI - Compositional analysis based on electron holography and a chemically sensitive reflection. AB - A method for compositional analysis of low-dimensional heterostructures is presented. The suggested procedure is based on electron holography and the exploitation of the chemically sensitive (0 0 2) reflection. We apply an off-axis imaging condition with the (0 0 2) beam strongly excited and centered on the optic axis. The first side band of the hologram is centered using an "empty" reference hologram obtained for a hole of the specimen. From the centered side band we use the phase of the central (0 0 0) and the amplitude of the (0 0 2) reflections to evaluate the local composition and the local specimen thickness in an iterative and self-consistent way. Delocalization effects that lead to a shift of the spatial information of (0 0 0) and (0 0 2) reflections are taken into account. The application of the procedure is demonstrated with an AlAs/GaAs(0 0 1) superlattice with a period of 5 nm. The concentration profiles obtained are discussed in relation to segregation. The measured segregation efficiency is R = 0.51 +/- 0.02. PMID- 11393452 TI - Structure determination and structure refinement of Al2CuMg precipitates by quantitative high-resolution electron microscopy. AB - The structure of the S-phase (Al2CuMg) precipitate in an Al matrix has been determined by using a combination of image processing, quantitative image comparison, and automatic refinement of imaging and structural parameters. A method for comparing images with unknown origin relationship while quickly estimating a possible translation of the origin is outlined. The optimization algorithm used in the structure determination utilizes space group symmetry, which is deduced from the crystal-zone axis and reduces the number of free parameters. PMID- 11393453 TI - The percept first, last, and always: the importance of microfacts in Rorschach analysis of the human movement response. PMID- 11393454 TI - Nostalgia, integration, validity issues, and generative power: some thoughts on Zygmunt A. Piotrowski. PMID- 11393455 TI - Honoring Zygmunt A. Piotrowski: memories and influence. PMID- 11393456 TI - A life fully lived, told in brief by his partner. PMID- 11393457 TI - The relationship of the Rorschach Schizophrenia Index to psychotic features in a child psychiatric sample. AB - In this study we investigated the reliability and validity of the Rorschach Schizophrenia Index (SCZI) from Exner's (1978, 1993) Comprehensive System for a sample of 413 child psychiatric inpatients by examining relationships with the Personality Inventory for Children-Revised (PIC-R) and chart diagnoses. Interscorer reliability and internal consistency were acceptable. Multivariate analyses of variance results revealed significantly different PIC-R profiles for those with and without elevated SCZI scores, with significant differences emerging on the PIC-R Psychosis (PSY) scale and 2 cognitive triad scales (Intellectual Screening and Development), which have been reported to be more frequently elevated in PIC-R profiles of children with psychotic disorders. Significant differences were found across SCZI groups for the PSY scale, Reality Distortion scale, reality testing critical items and chart diagnoses of psychotic disorder. Implications for clinical interpretation of the SCZI with children and issues for further research with this population are discussed. PMID- 11393458 TI - The impact of modified instructions on ego-level scores: a psychometric hazard or indication of optimal ego level? AB - Various modified instructions have different effects on the Washington University Sentence Completion Test for Ego Development (Loevinger, 1985). These effects were reviewed and 2 alternative explanations were explored: The "measurement unreliability explanation" versus the "optimal level explanation." Both explanations were systematically studied in 2 test-retest experiments with the Sentence Completion Test for Children and Youths (Westenberg, Treffers, & Drewes, 1998). The modified instructions were to make a favorable impression on the tester ("fake good" condition) or to complete the sentences in as adult and mature a manner as one can ("be mature" condition). Both experiments were conducted with 9- to 15-year-old children and adolescents (N = 127, 128). As was anticipated, neither the fake good nor the be mature condition yielded higher (or lower) reliability indexes as compared to the standard instructions, hence discounting the measurement unreliability explanation. Also as expected, the fake good condition did not yield significantly higher ego-level scores, whereas the be mature condition did yield significantly higher ego-level scores. The former instructions did not convey information relevant to the construct or measure of ego development, whereas the latter instructions did convey information relevant for raising ego-level scores. It is argued that the higher scores under the be mature instructions might reflect the respondents' "optimal" ego level (best functioning), whereas the ego-level score under the standard instructions might reflect their "functional" level (normal functioning). PMID- 11393459 TI - The Personality Inventory for Youth: validity and comparability of English and Spanish versions for regular education and juvenile justice samples. AB - In this study we examined the comparability of the Personality Inventory for Youth (PIY; Lachar & Gruber, 1993, 1995)--Spanish version to its English counterpart among bilingual Mexican American male and female high school students (n = 120), and among incarcerated bilingual Mexican American male adolescents (n = 52). Results indicated that language (English vs. Spanish) was not associated with a significant effect on the PIY scales or subscales for male adolescents in the juvenile justice facility, but was associated with a marginal effect for regular education adolescents; more specifically, regular education students completing the PIY in Spanish reported somewhat lower levels of symptoms than those completing the English version. Also, incarcerated males obtained significantly higher scores than regular education males on all 9 clinical and 19 out of 24 clinical subscales. Overall, the results of this study suggest that both language versions of the PIY may be applied usefully in the study and treatment of a variety of problems relevant to juvenile populations. Additional findings are discussed. PMID- 11393460 TI - The effects of MMPI-A T-score elevation on classification accuracy for normal and clinical adolescent samples. AB - In this investigation we examined the ability of the Minnesota Multiphasic Personality Inventory-Adolescent (MMPI-A; Butcher et al., 1992) to classify accurately both clinical and normal adolescents using 2 different T-score elevation ranges, T > or = 60 and T > or = 65, and using 2 different clinical base rates for the occurrence of psychopathology. A clinical base rate of 50% and 20%, respectively, were created by comparing a clinical sample of 203 adolescent inpatients with cooccurring substance abuse and psychiatric disorders with 2 subsamples from the MMPI-A normative group. These subsamples consisted of 203 adolescents matched for sex and age, and a larger subsample of 1,015 adolescents proportionately matched for sex and age, with the clinical group. Classification accuracy analyses revealed that although clinical base rate did affect the accurate classification of cases, a T-score cutoff of 65 resulted in higher levels of accurate classification overall while minimizing the misclassification of both clinical and normal cases. Implications of these findings for the recommended use of the MMPI-A "gray zone" are presented, and the relative areas of strength and weakness of the MMPI-A are reviewed in the identification and description of psychopathology. PMID- 11393461 TI - An evaluation of the usefulness of the MMPI-2 F(p) scale. AB - The Minnesota Multiphasic Personality Inventory-2 (MMPI-2) F(p) scale was developed by Arbisi and Ben-Porath (1995) by identification of 27 items endorsed by fewer than 20% of individuals in both normal and psychiatric samples. The F(p) scale was designed for applications in settings characterized by high base rates of serious psychopathology, such as psychiatric inpatient units, and is proposed as a useful scale in discriminating overreported protocols from those produced by patients with serious psychopathology. In this study we investigated the characteristics of this scale in a sample of 617 psychiatric inpatients who responded to the MMPI-2 under standard conditions, and 203 overreported protocols derived in research studies conducted with normal adult participants instructed to simulate various forms of serious psychopathology. Results of this study are consistent with prior reports of a relatively low frequency of item endorsement for F(p) scale items in psychiatric samples, and intercorrelations between the F(p) scale and the MMPI-2 basic clinical scales in clinical samples that are generally lower than those produced between either F or Fb and the basic clinical scales. However, this intercorrelational pattern between F(p) and the MMPI-2 basic scales was not as consistent for the overreported sample. Additionally, the F(p) scale appears to be effective in discriminating overreported from accurate MMPI-2 protocols, with some evidence that the optimal cutting scores for this and other MMPI-2 infrequency scales may differ as a function of gender. Finally, these findings do not show clear evidence of improved group prediction derived from the use of the F(p) scale in contrast to results obtainable through the use of the MMPI-2 F scale. PMID- 11393462 TI - MMPI-2 fake-bad scales: an attempted cross-validation of proposed cutting scores for outpatients. AB - Minnesota Multiphasic Personality Inventory-2 (MMPI-2) results were compared in 118 psychiatric outpatients given standard instructions, instructions to exaggerate their problems, instructions to feign a disorder they did not have, or instructions to feign global psychological disturbance. The groups were comparable on demographic, occupational and diagnostic characteristics as well as intake MMPI-2 results. Experimental MMPI-2 results showed that clinical scales were generally elevated in the feigning groups, with only modest differences across dissimulating instruction sets. The feigning groups had reliably higher scores than controls on all overreporting indexes examined, although no significant differences between feigning groups were present for overreporting indexes. Classification rates using previously proposed cutting scores for outpatients on individual feigning indexes showed near perfect specificity, but low to at best moderate sensitivity. Multiple regression analyses indicated that Gough's (1954) Dissimulation Scale (Ds2) was most strongly related to feigning status, and no other feigning scale contributed a significant increment in predictive power once Ds2 was entered. PMID- 11393463 TI - Semantic response consistency and protocol validity in structured personality assessment: the case of the NEO-PI-R. AB - In this study we tested the hypothesis that groups of NEO Personality Inventory Revised (NEO-PI-R; Costa & McCrae, 1992a) protocols identified as potentially invalid by an inconsistency scale (INC; Schinka, Kinder, & Kremer, 1997) would show reduced reliability and validity according to a series of psychometric tests. Data were obtained from 2 undergraduate student samples, a self-report group (n = 132) who provided NEO-PI-R self-ratings on 2 occasions separated by a 7- to 14-day interval and an informant group (n = 109) who provided ratings of well-known friends or relatives on 2 occasions separated by a 6 month interval. INC scores from the Time 1 protocols were used to divide these samples into low, moderate, and elevated inconsistency groups. In both samples, these 3 groups showed equivalent levels of reliability and validity as measured by: contingency coefficients for the 20 INC item responses across occasions; test-retest intraclass correlations of NEO-PI-R domain scores; convergent correlations with Goldberg's (1992) Bipolar Adjective Scale scores; and discriminant correlations between the 5 NEO-PI-R domain scores. The similarity of results across self report and informant assessment contexts provides additional evidence that semantic consistency approaches to assessing protocol validity may overestimate the prevalence of random or careless response behavior in standard administration conditions. Several theories are discussed that accommodate the existence of valid inconsistency in structured personality assessment. PMID- 11393464 TI - The Rorschach Suicide Constellation: assessing various degrees of lethality. AB - In this article we examine the relation between the Rorschach Comprehensive System's Suicide Constellation (S-CON; Exner, 1993; Exner & Wiley, 1977) and lethality of suicide attempts during the course of patients' hospitalization at the Austen Riggs Center (Stockbridge, MA). Patient records were rated as nonsuicidal (n = 37), parasuicidal (n = 37), or near-lethal (n = 30) based on the presence and lethality of self-destructive acts. Diagnostic efficiency statistics utilizing a cutoff score of 7 or more positive indicators successfully predicted which patients would engage in near-lethal suicidal activity relative to parasuicidal patients (overall correct classification rate [OCC] = .79), nonsuicidal inpatients (OCC = .79), and college students (OCC = .89). Although these predictions were influenced by relatively high base rates in the hospital population (14.5%), base rate estimates were calculated for other hypothetical populations revealing different prediction estimates that should be considered when judging the relative efficacy of the S-CON. Logistic regression analysis revealed that an S-CON score of 7 or more was the sole predictor of near-lethal suicide attempts among 9 psychiatric and demographic variables. PMID- 11393465 TI - Projective techniques at midcentury: a retrospective review of An introduction to projective techniques by Harold H. Anderson and Gladys L. Anderson. PMID- 11393466 TI - Medical physics education programs should include an in-depth course in statistics and mathematical modeling of human anatomy and physiology. PMID- 11393467 TI - Clinical use of electronic portal imaging: report of AAPM Radiation Therapy Committee Task Group 58. AB - AAPM Task Group 58 was created to provide materials to help the medical physicist and colleagues succeed in the clinical implementation of electronic portal imaging devices (EPIDs) in radiation oncology. This complex technology has matured over the past decade and is capable of being integrated into routine practice. However, the difficulties encountered during the specification, installation, and implementation process can be overwhelming. TG58 was charged with providing sufficient information to allow the users to overcome these difficulties and put EPIDs into routine clinical practice. In answering the charge, this report provides; comprehensive information about the physics and technology of currently available EPID systems; a detailed discussion of the steps required for successful clinical implementation, based on accumulated experience; a review of software tools available and clinical use protocols to enhance EPID utilization; and specific quality assurance requirements for initial and continuing clinical use of the systems. Specific recommendations are summarized to assist the reader with successful implementation and continuing use of an EPID. PMID- 11393468 TI - Detector line spread functions determined analytically by transport of Compton recoil electrons. AB - To achieve the maximum benefit of conformal radiation therapy it is necessary to obtain accurate knowledge of radiation beam penumbras based on high-resolution relative dosimetry of beam profiles. For this purpose there is a need to perform high-resolution dosimetry with well-established routine dosimeters, such as ionization chambers or diodes. Profiles measured with these detectors must be corrected for the dosimeter's nonideal response, caused by finite dimensions and, in the case of an ionization chamber, the alteration of electron transport and a contribution of electrons recoiled in the chamber wall and the central electrode. For this purpose the line spread function (LSF) of the detector is needed. The experimental determination of LSFs is cumbersome and restricted to the specific detector and beam energy spectrum used. Therefore, a previously reported analytical model [Med. Phys. 27, 923-934 (2000)] has been extended to determine response profiles of routine dosimeters: shielded diodes and, in particular, ionization chambers, in primary dose slit beams. The model combines Compton scattering of incident photons, the transport of recoiled electrons by Fermi Eyges small-angle multiple scattering theory, and functions to limit electron transport. It yields the traveling direction and the energy of electrons upon incidence on the detector surface. In the case of ionization chambers, geometrical considerations are then sufficient to calculate the relative amount of ionization in chamber air, i.e., the detector response, as a function of the detector location in the slit beam. In combination with the previously reported slit beam dose profiles, the LSF can then readily be derived by reconstruction techniques. Since the spectral contributions are preserved, the LSF of a dosimeter is defined for any beam for which the effective spectrum is known. The detector response profiles calculated in this study have been verified in a telescopic slit beam geometry, and were found to correspond to experimental profiles within 0.2 and 0.3 mm (full width at half-maximum) for a Wellhoefer IC15 chamber in a 6 and 25 MV-X x-ray beam, respectively. For a shielded diode these figures were found to be 0.2 and 0.1 mm, respectively. It is shown that a shielded diode in a primary beam needs only a small size-based correction of measured profiles. The effect of the LSF of an IC15 chamber on penumbra width has been determined for a set of model penumbras. The LSFs calculated by the application of the analytical model yield a broadening by 2 mm of a 3 mm wide penumbra (20%-80%). This is 0.5 mm (6 MV-X) to 1 mm (25 MV-X) smaller than found with the experimental LSFs. With a spatial correction based on the LSFs that were determined in this study, this broadening of up to 2 mm is eliminated, so that ionization chambers like the IC15 can be used for high-resolution relative dosimetry on a routine basis. PMID- 11393469 TI - Characterization of a commercial multileaf collimator used for intensity modulated radiation therapy. AB - The characteristics of a commercial multileaf collimator (MLC) to deliver static and dynamic multileaf collimation (SMLC and DMLC, respectively) were investigated to determine their influence on intensity modulated radiation therapy (IMRT) treatment planning and quality assurance. The influence of MLC leaf positioning accuracy on sequentially abutted SMLC fields was measured by creating abutting fields with selected gaps and overlaps. These data were also used to measure static leaf positioning precision. The characteristics of high leaf-velocity DMLC delivery were measured with constant velocity leaf sequences starting with an open field and closing a single leaf bank. A range of 1-72 monitor units (MU) was used providing a range of leaf velocities. The field abutment measurements yielded dose errors (as a percentage of the open field max dose) of 16.7+/-0.7% mm(-1) and 12.8+/-0.7% mm(-1) for 6 MV and 18 MV photon beams, respectively. The MLC leaf positioning precision was 0.080+/-0.018 mm (single standard deviation) highlighting the excellent delivery hardware tolerances for the tested beam delivery geometry. The high leaf-velocity DMLC measurements showed delivery artifacts when the leaf sequence and selected monitor units caused the linear accelerator to move the leaves at their maximum velocity while modulating the accelerator dose rate to deliver the desired leaf and MU sequence (termed leaf velocity limited delivery). According to the vendor, a unique feature to their linear accelerator and MLC is that the dose rate is reduced to provide the correct cm MU(-1) leaf velocity when the delivery is leaf-velocity limited. However, it was found that the system delivered roughly 1 MU per pulse when the delivery was leaf-velocity limited causing dose profiles to exhibit discrete steps rather than a smooth dose gradient. The root mean square difference between the steps and desired linear gradient was less than 3% when more than 4 MU were used. The average dose per MU was greater and less than desired for closing and opening leaf patterns, respectively, when the delivery was leaf-velocity limited. The results indicated that the dose delivery artifacts should be minor for most clinical cases, but limit the assumption of dose linearity when significantly reducing the delivered dose for dosimeter characterization studies or QA measurements. PMID- 11393470 TI - Dosimetric effects of contrast media for catheter-based intravascular brachytherapy. AB - The purpose of this work is to calculate the dose perturbations due to the presence of iodine contrast media in catheter-based intravascular brachytherapy (IVBT). The EGSnrc Monte Carlo system was used to perform the calculation for the two most commonly used beta and gamma sources (90Sr and 192Ir as well as for a variety of monoenergetic photon and electron sources. Various contrast media with cylindrical-shell shapes were considered. The dose perturbations due to the presence of iodine contrast media were found to be significant for IVBT. In the region outside of the contrast media (usually the treatment target region for IVBT), the dose is generally increased for the gamma source, while it is decreased for the beta source. In the presence of a 1.0-mm-thick common clinical contrast medium, the dose enhancement can be as high as 30% for the 192Ir source, while the dose can be reduced by up to 25% for the 90Sr source. The dose enhancement region for the gamma source extends approximately 1.0 mm away from the contrast medium, while the dose reduction for the beta source affects the entire target region. The dose perturbation depends on (1) the volume (thickness) of the contrast medium, (2) the iodine concentration in contrast media, and (3) the type and energy of radioactive source. The dose effects due to the contrast medium as well as the differences between beta and gamma sources, revealed in this study, should be recognized and may be considered in the dose prescription for intravascular brachytherapy. These effects should also be taken into account in analyzing the treatment outcome for those IVBT clinical trials involving a contrast medium. PMID- 11393471 TI - Monte Carlo-aided dosimetry of the Source Tech Medical Model STM1251 I-125 interstitial brachytherapy source. AB - We have used Monte Carlo photon transport simulations to calculate the dosimetric parameters of a new 125I seed, the Source Tech Medical Model STM125I source for interstitial brachytherapy. We followed the recommendations of the AAPM Task Group 43 and determined the following parameters: dose-rate constant, radial dose function, anisotropy function, anisotropy factor, and anisotropy constant. The recently (January 1999) revised National Institute of Standards and Technology I 125 standard for air-kerma strength calibration was taken into account as well as updated interaction cross-section data. The calculated dose-rate constant, when normalized to the simulated wide-angle, free-air chamber measurement of air-kerma strength, is 0.980 cGy h(-1) U(-1). The calculated radial dose function for the Model STM 1251 source is more penetrating than that of the model 6711 seed (by 18% at 5 cm distance), but agrees closely (within statistical errors) with that of the model 6702 seed up to distances of 10 cm. The STM125I source anisotropy functions indicate that its dose distribution is somewhat more anisotropic than that of the model 6702 and 6711 seeds at 1 cm distance but is comparable at larger distances. The Model STM125I anisotropy constant is very similar to that of the model 6711, 6702, and MED363I A/M seeds. PMID- 11393472 TI - Inverse planning anatomy-based dose optimization for HDR-brachytherapy of the prostate using fast simulated annealing algorithm and dedicated objective function. AB - An anatomy-based dose optimization algorithm is developed to automatically and rapidly produce a highly conformal dose coverage of the target volume while minimizing urethra, bladder, and rectal doses in the delivery of an high dose rate (HDR) brachytherapy boost for the treatment of prostate cancer. The dwell times are optimized using an inverse planning simulated annealing algorithm (IPSA) governed entirely from the anatomy extracted from a CT and by a dedicated objective function (cost function) reflecting clinical prescription and constraints. With this inverse planning approach, the focus is on the physician's prescription and constraint instead of on the technical limitations. Consequently, the physician's control on the treatment is improved. The capacity of this algorithm to represent the physician's prescription is presented for a clinical prostate case. The computation time (CPU) for IPSA optimization is less than 1 min (41 s for 142915 iterations) for a typical clinical case, allowing fast and practical dose optimization. The achievement of highly conformal dose coverage to the target volume opens the possibility to deliver a higher dose to the prostate without inducing overdosage of urethra and normal tissues surrounding the prostate. Moreover, using the same concept, it will be possible to deliver a boost dose to a delimited tumor volume within the prostate. Finally, this method can be easily extended to other anatomical sites. PMID- 11393473 TI - Absorbed dose estimates to structures of the brain and head using a high resolution voxel-based head phantom. AB - The purpose of this article is to demonstrate the viability of using a high resolution 3-D head phantom in Monte Carlo N-Particle (MCNP) for boron neutron capture therapy (BNCT) structure dosimetry. This work describes a high-resolution voxel-based model of a human head and its use for calculating absorbed doses to the structures of the brain. The Zubal head phantom is a 3-D model of a human head that can be displayed and manipulated on a computer. Several changes were made to the original head phantom which now contains over 29 critical structures of the brain and head. The modified phantom is a 85 x 109 x 120 lattice of voxels, where each voxel is 2.2 x 2.2 x 1.4 mm3. This model was translated into MCNP lattice format. As a proof of principle study, two MCNP absorbed dose calculations were made (left and right lateral irradiations) using a uniformly distributed neutron disk source with an 1/E energy spectrum. Additionally, the results of these two calculations were combined to estimate the absorbed doses from a bilateral irradiation. Radiobiologically equivalent (RBE) doses were calculated for all structures and were normalized to 12.8 Gy-Eq. For a left lateral irradiation, the left motor cortex receives the limiting RBE dose. For a bilateral irradiation, the insula cortices receive the limiting dose. Among the nonencephalic structures, the parotid glands receive RBE doses that were within 15% of the limiting dose. PMID- 11393474 TI - Toward clinical application of prompt gamma spectroscopy for in vivo monitoring of boron uptake in boron neutron capture therapy. AB - In boron neutron capture therapy (BNCT) the absorbed dose to the tumor cells and healthy tissues depends critically on the boron uptake. Pronounced individual variations in the uptake patterns have been observed for two boron compounds currently used in clinical trials. This implies a high uncertainty in the determination of the boron dose component. In the present work a technique known as prompt gamma spectroscopy (PGS) is studied that potentially can be used for in vivo and noninvasive boron concentration determination at the time of the treatment. The technique is based upon measurement of gamma rays promptly emitted in the 10B(n,alpha)7Li and 1H(n,gamma)2D reactions. The aim of this work is to prepare the present setup for clinical application as a monitor of boron uptake in BNCT patients. Therefore, a full calibration and a set of phantom experiments were performed in a clinical setting. Specifically, a nonuniform boron distribution was studied; a skin/ dura, a larger blood vessel, and tumor within a head phantom was simulated. The results show that it is possible to determine a homogeneous boron concentration of 5 microg/g within +/-3% (1 standard deviation). In the nonuniform case, this work shows that the boron concentration can be determined through a multistep measurement procedure, however, with a somewhat higher uncertainty (approximately 10%). The present work forms the basis for a subsequent clinical application of the PGS setup aimed at in vivo monitoring of boron uptake. PMID- 11393475 TI - In-phantom dosimetry for the 13C(d,n)14N reaction as a source for accelerator based BNCT. AB - The use of the 13C(d,n) 14N reaction at Ed=1.5 MeV for accelerator-based boron neutron capture therapy (AB-BNCT) is investigated. Among the deuteron-induced reactions at low incident energy, the 3C(d,n)14N reaction turns out to be one of the best for AB-BNCT because of beneficial materials properties inherent to carbon and its relatively large neutron production cross section. The deuteron beam was produced by a tandem accelerator at MIT's Laboratory for Accelerator Beam Applications (LABA) and the neutron beam shaping assembly included a heavy water moderator and a lead reflector. The resulting neutron spectrum was dosimetrically evaluated at different depths inside a water-filled brain phantom using the dual ionization chamber technique for fast neutrons and photons and bare and cadmium-covered gold foils for the thermal neutron flux. The RBE doses in tumor and healthy tissue were calculated from experimental data assuming a tumor 10B concentration of 40 ppm and a healthy tissue 10B concentration of 11.4 ppm (corresponding to a reported ratio of 3.5:1). All results were simulated using the code MCNP, a general Monte Carlo radiation transport code capable of simulating electron, photon, and neutron transport. Experimental and simulated results are presented at 1, 2, 3, 4, 6, 8, and 10 cm depths along the brain phantom centerline. An advantage depth of 5.6 cm was obtained for a treatment time of 56 min assuming a 4 mA deuteron current and a maximum healthy tissue dose of 12.5 RBE Gy. PMID- 11393476 TI - A neural network approach to breast cancer diagnosis as a constraint satisfaction problem. AB - A constraint satisfaction neural network (CSNN) approach is proposed for breast cancer diagnosis using mammographic and patient history findings. Initially, the diagnostic decision to biopsy was formulated as a constraint satisfaction problem. Then, an associative memory type neural network was applied to solve the problem. The proposed network has a flexible, nonhierarchical architecture that allows it to operate not only as a predictive tool but also as an analysis tool for knowledge discovery of association rules. The CSNN was developed and evaluated using a database of 500 nonpalpable breast lesions with definitive histopathological diagnosis. The CSNN diagnostic performance was evaluated using receiver operating characteristic analysis (ROC). The results of the study showed that the CSNN ROC area index was 0.84+/-0.02. The CSNN predictive performance is competitive with that achieved by experienced radiologists and backpropagation artificial neural networks (BP-ANNs) presented before. Furthermore, the study illustrates how CSNN can be used as a knowledge discovery tool overcoming some of the well-known limitations of BP-ANNs. PMID- 11393477 TI - Cone beam volume CT image artifacts caused by defective cells in x-ray flat panel imagers and the artifact removal using a wavelet-analysis-based algorithm. AB - The application of x-ray flat panel imagers (FPIs) in cone beam volume CT (CBVCT) has attracted increasing attention. However, due to a deficient semiconductor array manufacturing process, defective cells unavoidably exist in x-ray FPIs. These defective cells cause their corresponding image pixels in a projection image to behave abnormally in signal gray level, and result in severe streak and ring artifacts in a CBVCT image reconstructed from the projection images. Since a three-dimensional (3-D) back-projection is involved in CBVCT, the formation of the streak and ring artifacts is different from that in the two-dimensional (2-D) fan beam CT. In this paper, a geometric analysis of the abnormality propagation in the 3D back-projection is presented, and the morphology of the streak and ring artifacts caused by the abnormality propagation is investigated through both computer simulation and phantom studies. In order to calibrate those artifacts, a 2D wavelet-analysis-based statistical approach to correct the abnormal pixels is proposed. The approach consists of three steps: (1) the location-invariant defective cells in an x-ray FPI are recognized by applying 2-D wavelet analysis on flat-field images, and a comprehensive defective cell template is acquired; (2) based upon the template, the abnormal signal gray level of the projection image pixels corresponding to the location-invariant defective cells is replaced with the interpolation of that of their normal neighbor pixels; (3) that corresponding to the isolated location-variant defective cells are corrected using a narrow-windowed median filter. The CBVCT images of a CT low-contrast phantom are employed to evaluate this proposed approach, showing that the streak and ring artifacts can be reliably eliminated. The novelty and merit of the approach are the incorporation of the wavelet analysis whose intrinsic multi resolution analysis and localizability make the recognition algorithm robust under variable x-ray exposure levels between 30% and 70% of the dynamic range of an x-ray FPI. PMID- 11393479 TI - High-resolution polymer gel dosimetry by parameter selective MR-microimaging on a whole body scanner at 3T. AB - High dose variations across small spatial distances, as present in brachytherapeutic applications or radiosurgery and especially gamma-knife therapy, are difficult to quantify by standard dosimetry. We demonstrate the possibility to obtain planar spatial resolutions for dose imaging at pixel sizes below 200 microm within multislice parameter selective MR imaging on polymer gels. The sensitivity of the transversal and longitudinal relaxation time as well as diffusivity on dose is shown. High spatial resolution is achieved by parameter selective microimaging of polymer gels on a high-field (3 T) whole-body MR system equipped with a dedicated strong gradient system and a small probe head matched to the sample size. In addition to the spin-spin relaxation rate R2 = 1/T2 we investigate the sensitivity of the longitudinal relaxation rate R1 = 1/T1 and the diffusivity Dapp in acrylic polymer gels on irradiation up to dose levels of about 20 Gy. Dose images are obtained after calibration of the corresponding MR parameters by known dose levels of gamma irradiation. Also the MR-parameter T1 may be used for dose imaging. The impact of all of the three parameters T1, T2, and diffusivity on obtained signal intensities in irradiated regions has to be taken into account in nonoptimized pulse sequences. Further, very high spatial resolution imposes several restrictions on the evaluation of R2, which have to be considered for quantitative dosimetry. These restrictions are discussed in detail. We also demonstrate the importance of such a high spatial resolution in case of a set of differently sized gamma-knife stereotactic irradiation schemes. Gel dosimetry based on MR parameter selective microimaging represents a potent alternative for the detection of dose distributions characterized by steep dose gradients, typical in brachytherapeutic and radiosurgical applications. PMID- 11393478 TI - A Green's function approach to local rf heating in interventional MRI. AB - Current safety regulations for local radiofrequency (rf) heating, developed for externally positioned rf coils, may not be suitable for internal rf coils that are being increasingly used in interventional MRI. This work presents a two-step model for rf heating in an interventional MRI setting: (1) the spatial distribution of power in the sample from the rf pulse (Maxwell's equations); and (2) the transformation of that power to temperature change according to thermal conduction and tissue perfusion (tissue bioheat equation). The tissue bioheat equation is approximated as a linear, shift-invariant system in the case of local rf heating and is fully characterized by its Green's function. Expected temperature distributions are calculated by convolving (averaging) transmit coil specific absorption rate (SAR) distributions with the Green's function. When the input SAR distribution is relatively slowly varying in space, as is the case with excitation by external rf coils, the choice of averaging methods makes virtually no difference on the expected heating as measured by temperature change (deltaT). However, for highly localized SAR distributions, such as those encountered with internal coils in interventional MRI, the Green's function method predicts heating that is significantly different from the averaging method in current regulations. In our opinion, the Green's function method is a better predictor since it is based on a physiological model. The Green's function also elicits a time constant and scaling factor between SAR and deltaT that are both functions of the tissue perfusion rate. This emphasizes the critical importance of perfusion in the heating model. The assumptions made in this model are only valid for local rf heating and should not be applied to whole body heating. PMID- 11393480 TI - The thermal characteristics of different diodes on in vivo patient dosimetry. AB - Diode sensitivity variations with temperature (SVWT) have been reported to vary from small negative values up to 0.6% per degrees C. Thus it is possible for diode calibration factors established at room temperature (approximately 20 degrees C) to yield errors in the range of -1% to +9% when diodes are placed on a patient's skin (approximately 30 degrees C) for in vivo entrance dose measurements. In this study we simulated several skin temperatures using a temperature-controlled aluminum surface in contact with a section of Bolus. The internal temperatures of several diodes with different buildup thickness were monitored as a function of time when placed in contact with the heated bolus. Our results indicate that for different combinations of room temperature (18 degrees C-23 degrees C) and patient skin temperature (28 degrees C-34 degrees C) diodes reached 90% of their equilibrium temperature within 3-5 min. In addition, the range of typical skin temperatures was determined by measurements performed on a number of actual patients under clinical conditions. Based on the results of our experiments a protocol was developed to minimize the temperature based errors for in vivo dosimetry. PMID- 11393481 TI - Determination of guidance levels of dose for diagnostic radiography in Taiwan. AB - The International Atomic Energy Agency has recommended guidance levels of dose for diagnostic radiography for a typical adult patient. These levels were intended to act as thresholds to trigger investigations or corrective actions in ensuring optimized protection of patients and maintaining appropriate levels of good practice. Since guidance levels should be derived from wide scale surveys of exposure factors performed in individual hospitals, a national survey was conducted recently in Taiwan to collect these factors for the most frequent radiographic procedures. A total of 276 completed questionnaires were received and analyzed. In the questionnaire, respondents were asked to check those projections that were routinely performed in their department and to report machine data, patient data, output measurements, and technical factors including kVp, mAs, focus-to-film distance, table-to-film distance, aluminum filtration, and focal spot size. Based on the survey data, entrance skin exposures in air, i.e., free air exposures at the point of intersection of the x-ray central beam with the entrance surface of the patient, were estimated using the RADCOMP program. Entrance surface doses to air and tissue with backscatter were then evaluated by the application of the exposure-dose conversion factor and the backscatter factor obtained from TLD measurements and Monte Carlo simulations. Guidance levels were determined from survey results on the entrance surface dose based on optimization considerations involving the cost-effectiveness analysis. Except for chest PA and LAT and skull LAT procedures, all guidance levels derived in this work are less than those recommended by the International Atomic Energy Agency. Survey data and guidance levels were provided to the national authorities to help them develop quality control and radiation protection programs for medical exposures. PMID- 11393482 TI - Response to 'comment on "Functional fitting of interstitial brachytherapy dosimetry data recommended by the AAPM Radiation Therapy Committee Task Group 43" [Med. Phys., 26, 153-160 (1999)] and "Fitting and benchmarking of dosimetry data for new brachytherapy sources". PMID- 11393483 TI - Influenza B virus outbreak on a cruise ship--Northern Europe, 2000. AB - During June 23-July 5, 2000, an outbreak of respiratory illnesses occurred on the MS Rotterdam (Holland America Line & Windstar Cruises) during a 12-day Baltic cruise from the United Kingdom to Germany via Russia. The ship carried 1311 passengers, primarily from the United States, and 506 crew members from many countries. Although results of rapid viral testing for influenza A and B viruses were negative, immunofluorescence staining and viral culture results implicated influenza B virus infection as the cause of the outbreak. This report summarizes the findings of the outbreak investigation conducted by the ship's medical department and describes the measures taken to control the outbreak. Travelers at high risk for complications of influenza who were not vaccinated with influenza vaccine during the preceding fall or winter should consider receiving influenza vaccine before travel with large tourist groups at any time of year or to certain regions of the world. PMID- 11393484 TI - Blood and hair mercury levels in young children and women of childbearing age- United States, 1999. AB - Mercury (Hg), a heavy metal, is widespread and persistent in the environment. Exposure to hazardous Hg levels can cause permanent neurologic and kidney impairment. Elemental or inorganic Hg released into the air or water becomes methylated in the environment where it accumulates in animal tissues and increases in concentration through the food chain. The U.S. population primarily is exposed to methylmercury by eating fish. Methylmercury exposures to women of childbearing age are of great concern because a fetus is highly susceptible to adverse effects. This report presents preliminary estimates of blood and hair Hg levels from the 1999 National Health and Nutrition Examination Survey (NHANES 1999) and compares them with a recent toxicologic review by the National Research Council (NRC). The findings suggest that Hg levels in young children and women of childbearing age generally are below those considered hazardous. These preliminary estimates show that approximately 10% of women have Hg levels within one tenth of potentially hazardous levels indicating a narrow margin of safety for some women and supporting efforts to reduce methylmercury exposure. PMID- 11393485 TI - Progress toward poliomyelitis eradication--Afghanistan, 1999-2000. AB - In 1988, the World Health Assembly of the World Health Organization (WHO) resolved to eradicate poliomyelitis globally by 2000. During the same year, the Eastern Mediterranean Region (EMR) of WHO passed a resolution to join the global initiative. Since then, substantial progress has been made worldwide and in EMR member countries. Afghanistan, with ongoing civil conflict, initiated polio eradication activities in 1994. Since then, a countrywide surveillance system for acute flaccid paralysis (AFP) was established and National Immunization Days (NIDs) were implemented. This report summarizes the achievements toward polio eradication in Afghanistan during 1999-2000. PMID- 11393486 TI - Trends in screening for colorectal cancer--United States, 1997 and 1999. AB - Colorectal cancer is the second leading cause of cancer-related death in the United States. An estimated 135,400 new cases and 56,700 deaths from colorectal cancer are expected during 2001. Since the mid-1990s, national guidelines have recommended that persons aged >50 years at average risk for colorectal cancer should have screening tests regularly. To estimate rates for the use of colorectal cancer screening tests and to evaluate trends in test use, CDC analyzed data from the 1999 Behavioral Risk Factor Surveillance System (BRFSS) on the use of a home administered fecal occult blood test (FOBT) and sigmoidoscopy/colonoscopy, and then compared them with similar data from 1997. The findings in this report indicate that the proportion of the U.S. population that has been screened remains low. In 1999, 44% of BRFSS respondents reported receiving FOBT and/or sigmoidoscopy/colonoscopy within the recommended period compared with approximately 41% reporting FOBT and/or sigmoidoscopy/proctoscopy within the recommended period in 1997. Efforts to address barriers and to promote the use of colorectal cancer screening should be intensified. PMID- 11393487 TI - Physical activity trends--United States, 1990-1998. AB - Physical activity is associated with numerous health benefits, and increased participation in various types of leisure-time physical activity had been encouraged during the 1990s. To determine national estimates of leisure-time physical activity during 1990-1998, data were obtained from the Behavioral Risk Factor Surveillance System (BRFSS). This report summarizes the results of that analysis, which indicate that leisure-time physical activity trends have remained unchanged. PMID- 11393488 TI - Sudden death in a traveler following halofantrine administration--Togo, 2000. AB - On July 17, 2000, a previously healthy 22-year-old U.S. student collapsed and died suddenly while leading a teenage exchange group in West Africa. This report summarizes the results of the investigations of this incident, which implicate use of halofantrine for treatment of malaria as the cause of death. Travelers should be warned that halofantrine treatment may be dangerous in persons with cardiac abnormalities or in those taking mefloquine for malaria prophylaxis. PMID- 11393489 TI - Primary and secondary syphilis--United States, 1999. AB - In October 1999, CDC, in collaboration with other federal partners, launched the National Plan to Eliminate Syphilis in the United States. In 1998, Congress initiated funding for the syphilis elimination effort. Syphilis elimination is defined as the absence of sustained transmission (i.e., no transmission after 90 days of the report of an imported index case). The national goal for syphilis elimination is to reduce primary and secondary (P&S) syphilis to <1000 cases (rate: 0.4 per 100,000 population) and to increase the number of syphilis-free counties to 90% by 2005. To describe the epidemiology of syphilis in the United States, CDC analyzed notifiable disease surveillance data for 1999. This report summarizes the results of that analysis, which indicate that, in 1999, P&S syphilis declined to a rate of 2.5 cases per 100,000 population, the lowest rate ever reported, and that syphilis transmission increasingly is concentrated in a few geographic areas. PMID- 11393490 TI - Outbreak of syphilis among men who have sex with men--Southern California, 2000. AB - Syphilis is a sexually transmitted disease (STD) caused by infection with the spirochete Treponema pallidum, and like other genital ulcer diseases, syphilis enhances the transmission of human immunodeficiency virus (HIV). During the 1990s, syphilis occurred predominantly among heterosexual blacks in the South and in large cities. However, recent outbreaks of syphilis have occurred among men who have sex with men (MSM). A large syphilis outbreak occurred among MSM during January-July 2000 in southern California. During the outbreak period, the proportion of primary and secondary (P&S) syphilis cases among MSM increased to 51% from 26% for the same period in 1999. This report summarizes the findings of an investigation of this syphilis outbreak, which indicate a substantial increase in the number of syphilis cases among MSM, many of whom are HIV-positive. These data suggest that concern about HIV infection may be declining among MSM and emphasize the importance of strengthening efforts to prevent HIV infection in this population in the United States. PMID- 11393491 TI - Prevalence of disabilities and associated health conditions among adults--United States, 1999. AB - In the United States, the number of persons reporting disabling conditions increased from 49 million during 1991-1992 to 54 million during 1994-1995. During 1996, direct medical costs for persons with disability were $260 billion. Surveillance of disability prevalence and associated health conditions is useful in setting policy, anticipating the service needs of health systems, assisting state programs, directing health promotion and disease prevention efforts, and monitoring national health objectives. The U.S. Bureau of the Census and CDC analyzed data from the Survey of Income and Program Participation (SIPP) to determine national prevalence estimates of adults with disabilities and associated health conditions. This report summarizes findings of that analysis, which indicate that disability continues to be an important public health problem, even among working adults, and arthritis or rheumatism, back or spine problems, and heart trouble/hardening of the arteries remain the leading causes. Better health promotion and disease prevention may reduce the prevalence of disability-associated health conditions. PMID- 11393492 TI - Hartmut Michel--Nobel laureate in chemistry. PMID- 11393493 TI - The utility of PSA doubling time to monitor prostate cancer recurrence. PMID- 11393494 TI - TNF-alpha inhibition: the need for a tumor necrosis factor thermostat. PMID- 11393495 TI - PSA doubling time as a predictor of clinical progression after biochemical failure following radical prostatectomy for prostate cancer. AB - OBJECTIVES: To characterize the clinical progression of disease in men who have undergone prostatectomy for clinically localized prostate cancer and have postoperative biochemical failure (elevated prostate-specific antigen [PSA] level) and to identify predictors of clinical disease progression, including the possible effect of PSA doubling time (PSADT). PATIENTS AND METHODS: Between 1987 and 1993, 2809 patients underwent radical retropubic prostatectomy for clinically localized (< or =T2) disease. In our database, all patients with postoperative biochemical failure (PSA level > or =0.4 ng/mL) were identified. The PSADT was estimated using log linear regression on all PSA values (excluding those values determined after administration of hormonal therapy) within 15 months after biochemical failure. All patients had regular PSA measurements from the time of surgery through the follow-up period. Systemic progression (SP) was defined as evidence of metastatic disease on a bone scan. Local recurrence (LR) was defined on the basis of digital rectal examination, transrectal ultrasonography, and biopsy. The SP-free survival and LR/SP-free survival (survival free of both LR and SP) after biochemical failure was estimated with use of the Kaplan-Meier method. Patients with prostate cancer treatment after biochemical failure had their follow-up censored from this study at the time of treatment. RESULTS: Postoperative biochemical failure occurred in 879 men (31%). The mean follow-up from time of biochemical failure was 4.7 years (range, 0.5-11 years). The mean time to biochemical failure was 2.9 years (median, 2.4 years). The overall mean SP-free survival from time of biochemical failure was 94% and 91% at 5 and 10 years, respectively. The mean LR/SP-free survival was 64% and 53% at 5 and 10 years, respectively. By using univariate analysis on the 587 patients with PSADT data, significant risk factors for SP were PSADT (P<.001) and pathologic Gleason score (P=.005); for LR/SP, significant risk factors included PSADT (P<.001) and pathologic Gleason score (P<.001). In multivariate Cox models analysis, only PSADT remained a significant risk factor for both SP and LR/SP (P<.001). Mean 5 year SP-free survival was 99%, 95%, 93%, and 64% for patients with PSADT of 10 years or longer, 1.0 to 9.9 years, 0.5 to 0.9 year, and less than 0.5 year, respectively; the respective mean LR/SP-free survivals were 87%, 62%, 46%, and 38%. The percentage of patients with PSADT of less than 0.5 year was considerably higher if the type of first clinical event was SP (48%) compared with LR (18%) (P<.001). CONCLUSIONS: For patients who have undergone radical prostatectomy, a rising PSA level suggests evidence of residual or recurrent prostate cancer. Many men remain free of clinical disease for an extended time after biochemical failure following radical prostatectomy for clinically localized prostate cancer. The PSADT appears to be an important predictor of SP and also of any clinical progression (local or systemic). These data may be useful when counseling men regarding the timing of adjuvant therapies. PMID- 11393496 TI - Changes in testosterone, cortisol, and estradiol levels in men becoming fathers. AB - OBJECTIVE: To quantify longitudinally steroid hormone (testosterone, cortisol, and estradiol) concentrations in men becoming fathers for the first time ("dads"). SUBJECTS AND METHODS: Volunteer study subjects were recruited from first-trimester prenatal classes in Kingston, Ontario, in February 1999. Twenty three dads provided saliva samples from recruitment through 3 months after the birth of their children. Fourteen men who were not fathers were recruited from the general population to serve as age-matched controls for season and time of day. Estradiol, testosterone, and cortisol levels were quantified. RESULTS: After controlling for effects of time of day and season, dads had lower mean +/- SE testosterone (6.5+/-0.7 vs 10.0+/-0.9 ng/dL; P<.005) and cortisol (morning values, 0.30+/-0.05 vs 0.53+/-0.05 microg/dL; P<.005) concentrations, a higher proportion of samples with detectable estradiol concentrations (68% [308/454] vs 57% [87/154]; P=.01), and higher estradiol concentrations in those detectable samples (3.81+/-0.09 pg/mL [13 dads] vs 3.26+/-0.11 pg/mL [9 controls]; P<.002) than did control men. Within 10 individual dads with frequent samples before and after the birth, the percentage of samples with detectable estradiol was lower during the month before the birth than during the month after (51% vs 71%; P=.02), and cortisol concentration was increased in the week before the birth (to a mean of 0.16 microg/dL). In each of 13 dads providing frequent samples, testosterone concentration and variance were low immediately after the birth (no change from previous levels in 5, decrease after prebirth increase in 3, and decrease relative to all other times in 5). CONCLUSIONS: In this population of Canadian volunteers attending prenatal classes, expectant fathers had lower testosterone and cortisol levels and a higher proportion of samples with detectable estradiol concentrations than control subjects. Individual patterns of testosterone variance relative to the birth and estradiol and cortisol concentrations immediately before the birth may be worthy of further investigation. The physiologic importance of these hormonal changes, if any, is not known. However, they are hormones known to influence maternal behavior. PMID- 11393497 TI - Incidence of physician-diagnosed primary Sjogren syndrome in residents of Olmsted County, Minnesota. AB - OBJECTIVES: To estimate the incidence of physician-diagnosed primary Sjogren syndrome (SS) among residents of Olmsted County, Minnesota, in the setting of usual medical care and to determine how often objective criteria are available in the medical records of such patients. PATIENTS AND METHODS: We reviewed all medical records of residents in Olmsted County with physician-diagnosed SS from 1976 to 1992 to determine whether they had undergone objective tests for keratoconjunctivitis sicca, salivary dysfunction, or serologic abnormality. Confounding illnesses were excluded. To identify misclassified cases, all records from patients with xerostomia or keratoconjunctivitis sicca were also reviewed. The average annual SS incidence rates were calculated by considering the entire population to be at risk. RESULTS: Of 75 patients with onset of SS during the study period, 53 had primary SS. All patients were white, 51 (96.2%) were women, and the mean +/- SD age was 59+/-15.8 years. The age- and sex-adjusted annual incidence was 3.9 per 100,000 population (95% confidence interval, 2.8-4.9) for patients with primary SS. Eleven patients (20.8%) with physician-diagnosed SS had no documentation of objective eye, mouth, or laboratory abnormalities. Objective evaluations performed most frequently were laboratory and ocular tests and least often were investigations of xerostomia. CONCLUSIONS: The average annual incidence rate for physician-diagnosed primary SS in Olmsted County is about 4 cases per 100,000 population. These data probably underestimate the true incidence because they are based on usual medical care of patients with SS in a community setting, rather than on a case-detection survey. In the future, a true incidence may be possible with a higher index of suspicion, greater attention to objective tests, and increased awareness of new classification criteria for SS. For epidemiological studies based on existing data, application of current criteria may not be feasible, and consensus on criteria for such studies would be useful. PMID- 11393498 TI - Use of a transurethral microwave thermotherapeutic device with permanent pacemakers and implantable defibrillators. AB - OBJECTIVE: To determine whether a device (Urologix Targis system) used for transurethral microwave treatment interferes with sensing, pacing, and arrhythmia detection by permanent pacemakers and implantable cardioverter-defibrillators (ICDs). METHODS: We tested 13 pacemakers in both bipolar and unipolar sensing configurations and 8 ICDs in vitro. Pacemakers and ICDs were programmed to their most sensitive settings. Energy outputs of the microwave device were typical of those used clinically. The probe of the microwave device was anchored 1.2 cm from the pacemaker or ICD being tested. RESULTS: No sensing, pacing, or arrhythmic interactions were noted with any ICD or any pacemaker programmed to the bipolar configuration. One pacemaker (Guidant Vigor 1230) showed intermittent tracking when programmed to the unipolar configuration. CONCLUSIONS: Most patients with permanent pacemakers or ICDs can safely undergo transurethral microwave therapy using the device tested. Pacemakers and ICDs should be programmed to the bipolar configuration (if available) during therapy. The pacemaker or ICD should be interrogated before and after therapy to determine whether programming changes occurred as a result of treatment. However, our findings suggest that this is unlikely. PMID- 11393499 TI - Patient satisfaction associated with correct identification of physician's photographs. AB - OBJECTIVE: To determine whether placement of photographs of physicians in hospital rooms improves patients' satisfaction with their medical care. PATIENTS AND METHODS: This is a prospective, controlled study of 224 patients admitted to general internal medicine services in a teaching hospital. The intervention consisted of photographs (8 x 10 in) of attending and resident physicians displayed in the patients' rooms. Before dismissal, patients completed a survey that required them to match names with photographs of physician caregivers and included patient satisfaction questions. The primary outcome was whether patients who had photographs in their hospital room would correctly identify more physicians than those with no photographs in their room. RESULTS: The presence of photographs on the hospital wall was associated with a significant improvement in the number of physicians identified correctly (odds ratio [OR], 1.83; 95% confidence interval [CI], 1.47-2.27; P<.001). The percentage of physicians that patients identified by correctly matching their physicians' names to their photographs was significantly associated with satisfaction with physician responsiveness (OR, 1.19; 95% CI, 1.01-1.40; P=.03) and with the way in which physicians addressed questions regarding medical care (OR, 1.23; 95% CI, 1.05 1.44; P=.05). CONCLUSIONS: Patients who had photographs of their physicians on the wall of their hospital room were able to identify correctly a larger number of physicians on their team compared with patients who had no photographs. Patient satisfaction was related to the number of physicians' photographs that patients could identify correctly. PMID- 11393500 TI - Gycemic control, mealtime glucose excursions, and diabetic complications in type 2 diabetes mellitus. AB - Type 2 diabetes mellitus is a heterogeneous disorder characterized by 2 pathogenic defects, impaired insulin secretion and insulin resistance. The resultant hyperglycemia causes microvascular and macrovascular complications that increase morbidity and mortality in patients with diabetes mellitus. Optimum glycemic control in patients with type 1 and type 2 diabetes mellitus prevents the development of microvascular disease and, to a lesser extent, macrovascular disease. Prandial hyperglycemia may be an independent risk factor for the development of diabetic complications. This article reviews the pathophysiologic mechanisms of glucose metabolism and describes the results of epidemiological and interventional studies that have demonstrated the association of acute and chronic hyperglycemia with the development of diabetic complications. The American Diabetes Association has defined diagnostic and treatment goals for diabetes mellitus, striving to achieve near-normal glycemic control to delay or prevent the development of diabetic complications. A number of oral antidiabetic agents and insulins are currently available for the treatment of type 2 diabetes mellitus in the United States. These agents target fasting and postmeal plasma glucose levels to improve glycemic control. Alone or in combination, these agents have enhanced the clinical approaches to treating diabetes mellitus. PMID- 11393501 TI - Involvement of oxidation-sensitive mechanisms in the cardiovascular effects of hypercholesterolemia. AB - Hypercholesterolemia is a common clinical metabolic and/or genetic disorder that promotes functional and structural vascular wall injury. The underlying mechanisms for these deleterious effects involve a local inflammatory response and release of cytokines and growth factors. Consequent activation of oxidation sensitive mechanisms in the arterial wall, modulation of intracellular signaling pathways, increased oxidation of low-density lipoprotein cholesterol, and quenching of nitric oxide can all impair the functions controlled by the vascular wall and lead to the development of atherosclerosis. This cascade represents a common pathological mechanism activated by various cardiovascular risk factors and may partly underlie synergism among them as well as the early pathogenesis of atherosclerosis. Antioxidant intervention and restoration of the bioavailability of nitric oxide have been shown to mitigate functional and structural arterial alterations and improve cardiovascular outcomes. Elucidation of the precise nature and role of early transductional signaling pathways and transcriptional events activated in hypercholesterolemia in children and adults, including mothers during pregnancy, and understanding their downstream effects responsible for atherogenesis may help in directing preventive and interventional measures against atherogenesis and vascular dysfunction. PMID- 11393502 TI - Images and reflections from Mayo Clinic heritage. PMID- 11393503 TI - A guide to the diagnosis and treatment of occupational asthma. AB - Occupational asthma is the most prevalent form of occupational lung disease in industrialized nations. As increasing numbers of new chemicals are produced and new manufacturing processes are introduced, the variety of environments in which individuals may become exposed to respiratory sensitizers and irritants makes diagnosing and treating this illness even more challenging. In addition to adverse pulmonary effects, the diagnosis of occupational asthma may bring with it negative social and financial implications that may ultimately affect the patient's quality of life. For this reason, it is important for clinicians to recognize work-related respiratory symptoms early on in their course, maintain a high clinical suspicion for an occupational cause in the diagnostic work-up of asthma, and have a high degree of certainty in the diagnosis. While a number of classification schemes have been proposed to simplify the diagnostic approach to occupational asthma, the inciting factors typically involve sensitization (often by an IgE mechanism), direct airway inflammation, various pharmacologic responses, or irritant reflex pathways. Clinicians must first document the presence of asthma, then establish a relationship between asthma and the work place. The occupational history is the key diagnostic tool, and clinical suspicions may be evaluated further by serial peak expiratory flow measurements, nonspecific hypersensitivity challenges with histamine or methacholine, collaboration with industrial/occupational hygienists to obtain workplace exposure measurements, and specific challenge testing at tertiary referral centers providing specialized laboratories. Removal from the inciting exposure is the mainstay of therapy, and pharmacologic treatment of patients with occupational asthma is similar to the treatment of patients with other forms of asthma. PMID- 11393504 TI - Detection and evaluation of a palpable breast mass. AB - The discovery of a breast mass, either self-detected or identified by a clinician, is a common, often distressing occurrence for many women. Although most detected breast masses are benign, every woman presenting with a breast mass should be evaluated to exclude or establish a diagnosis of cancer. This article provides a succinct overview of normal breast anatomy and reviews common causes of breast masses. The role of the clinical breast examination is discussed, and an algorithm is provided for optimal utilization of available tools in the diagnostic evaluation of a breast mass. The evaluation should be performed expeditiously and the results communicated promptly to the patient. Regardless of the age of the woman, a clinically suspicious mass must be evaluated even if findings on a mammogram are normal. PMID- 11393505 TI - 49-year-old woman with acute abdominal pain and nausea. PMID- 11393506 TI - Fatal sepsis in a patient with rheumatoid arthritis treated with etanercept. AB - Patients with long-standing, severe, erosive rheumatoid arthritis who have extra articular manifestations and have undergone joint replacement surgery are at increased risk for serious infection and premature mortality. New therapies, including cytokine antagonists, hold great promise for improving the course of rheumatoid arthritis. However, they have powerful anti-inflammatory effects that may mask symptoms of serious infection. We report a case of fatal pneumococcal sepsis occurring in a 37-year-old woman with rheumatoid arthritis treated with the tumor necrosis factor antagonist etanercept and suggest management strategies for early detection and management of this complication. PMID- 11393507 TI - Extension of a pulmonary blastoma into the left atrium. AB - Pulmonary tumors of embryonic origin are rare, and pulmonary blastomas are probably the most uncommon. A thorough literature search disclosed no previous reports of extension of this type of tumor into the heart. We describe a patient whose initial clinical presentation suggested an obstructive left atrial mass; however, clinical and histologic findings indicated the mass was a tumor that originated from a pulmonary blastoma that extended into the left atrium through a pulmonary vein. The unique aspect of this case is that the patient's symptoms were related to the obstructive effects of the atrial mass, not to the primary pulmonary tumor. PMID- 11393508 TI - Iliac arteriovenous fistula with renal insufficiency, ascites, hepatomegaly, and abnormal liver test results. AB - Arteriovenous fistula presents rarely with liver involvement. A 73-year-old man had new-onset ascites, hepatomegaly, and abnormal liver and renal function test results. An abdominal computed tomogram revealed a 7.6-cm internal iliac aneurysm but no other abnormality to account for his ascites. An aortogram demonstrated a 1.5-cm internal iliac arteriovenous fistula that subsequently was repaired, leading to resolution of his symptoms and laboratory abnormalities. High-output cardiac failure should be considered in the differential diagnosis of patients with new-onset massive ascites, hepatomegaly, and liver test abnormalities. PMID- 11393510 TI - The latest hype on Hyp-O-glycosylation codes. AB - Contemporary glycobiology reflects the intense interest in glycoproteins and their biological roles. Addition of saccharides by N- or O-glycosylation is precise rather than random and forms a uniquely interactive molecular surface. We designate these well conserved glycomotifs as glycomodules to emphasize their functional significance. Thus, elucidation of the glycosylation codes that determine saccharide addition is a significant goal. The focus here is on the Hyp O-glycosylation of cell wall proteins. This involves two consecutive posttranslational modifications, proline hydroxylation and glycosylation. Peptide sequence rather than conformation seems to determine these modifications. Hyp glycosylation occurs in two distinct modes: Hyp arabinosylation and Hyp galactosylation. The Hyp contiguity hypothesis predicts arabinosylation of contiguous Hyp residues and galactosylation of clustered non-contiguous Hyp. Elucidation of Hyp glycosylation codes involves the design and expression of putative glycomotifs as simple repetitive peptides. Thus, repetitive (Ser-Hyp), directed Hyp galactosylation resulting in the exclusive addition of arabinogalactan polysaccharide to all the non-contiguous Hyp residues. and a new AGP. Another repetitive peptide from gum arabic glycoprotein, containing both contiguous and non-contiguous Hyp, directed both modes of Hyp glycosylation. Furthermore, expression of the (Ser-Hypx)n series confirmed the arabinosylation of contiguous Hyp. Thus, the Hyp contiguity hypothesis is a useful predictive tool in the functional genomics toolbox. PMID- 11393509 TI - Bupropion-induced erythema multiforme. AB - The high rate of dermatologic adverse effects associated with bupropion use may extend to its sustained-release preparation, currently prescribed extensively for smoking cessation as well as for treatment of depressive conditions. We report what we believe to be the first case, in a 31-year-old woman, of erythema multiforme after administration of sustained-release bupropion (Wellbutrin SR) for treatment of depression. This report emphasizes that prescribers must aggressively follow up their patients who have rashes or urticaria, discontinuing the medication as soon as erythema multiforme is suspected and watching closely for the emergence of potentially life-threatening dermatologic conditions. PMID- 11393511 TI - Phytoecdysteroids: biological aspects. AB - Phytoecdysteroids are a family of about 200 plant steroids related in structure to the invertebrate steroid hormone 20-hydroxyecdysone. Typically, they are C27, C28 or C29 compounds possessing a 14alpha-hydroxy-7-en-6-one chromophore and A/B cis ring fusion (5beta-H). In the present review, the distribution, biosynthesis, biological significance and potential applications of phytoecdysteroids are summarised. PMID- 11393512 TI - Enzymatic halogenation of flavanones and flavones. AB - The whole cells and the chloroperoxidase enzyme of Caldariomyces fumago were capable of halogenating the flavanones, naringenin and hesperetin, at C-6 and C-8 in the presence of either Cl- or Br-. However, they did not act on other test flavones. The biohalogenated products of naringenin and hesperetin were isolated and found to be identical to those obtained from chemical reactions using molecular halogen and hypohalous acid. PMID- 11393513 TI - Purification and characterization of isoforms of beta-galactosidases in mung bean seedlings. AB - Five isoforms of beta-galactosidase (EC 3.2.1.23), designated as beta galactosidases I-V, were isolated from five-day-old mung bean (Vigna radiata) seedlings. Beta-galactosidases II and III were purified to electrophoretic homogeneity by a procedure involving acid precipitation, ammonium sulfate fractionation, chromatography on diethylaminoethyl-cellulose (DEAE-Cellulose) and con A-Sepharose. and chromatofocusing. Beta-galactosidases I, II and III have the same molecular mass of 87 kDa. comprising two nonidentical subunits with molecular masses of 38 and 48 kDa, while beta-galactosidases IV and V have molecular masses of 45 and 73 kDa, respectively. All the enzymes were active against p-nitrophenyl-beta-D-galactoside, and to a lesser extent, p-nitrophenyl alpha-L-arabinoside and p-nitrophenyl-beta-D-fucoside. The enzymes were inhibited by D-galactono-1,4-lactone, D-galactose, Hg2+, Ag+ and sodium dodecyl sulfate (SDS). Beta-galactosidases I, II and III were shown to be competitively inhibited by either D-galactono-1, 4-lactone or D-galactose. Isoforms I, II and III have a common optimal pH of 3.6, while isoforms IV and V have pH optima at 3.8 and 4.0, respectively. Isoelectric points of isoforms I, II and III were 7.7, 7.5 and 7.3, respectively. Double immunodiffusion analysis indicated that beta-galactosidases I, II, III and V are immunologically similar to each other, while beta galactosidase IV shares partially identical antigenic determinants with the other four isoforms. The purified beta-galactosidases II and III were capable of releasing D-galactose residue from the hemicellulose fraction isolated from mung bean seeds. PMID- 11393514 TI - Biosynthesis of 2-aminobenzaldehyde in flowers of Robinia pseudoacacia and Philadelphus coronarius. AB - Feeding experiments with 13C- and fluorine-labelled precursors were performed to reveal the biosynthesis of 2-aminobenzaldehyde in flowers of Robinia pseudoacacia and Philadelphus coronarius. The results are in agreement with the transformation of anthranilic acid to indole followed by oxidative ring opening and hydrolysis of the resulting N-formyl-2-aminobenzaldehyde. This route differs from that observed in Hebeloma sacchariolens (Basidiomycetes) in which anthranilic acid is directly reduced to 2-aminobenzaldehyde. PMID- 11393515 TI - Ginsenoside production in different phenotypes of Panax ginseng transformed roots. AB - Transformed roots were obtained after the inoculation of sterile root discs of Panax ginseng C.A. Meyer with Agrobacterium rhizogenes A4. The established hairy root lines displayed three morphological phenotypes when cultured on hormone-free liquid Schenk and Hildebrandt medium. Most of the cultures showed the characteristic traits of hairy roots (HR-M), while others were either callus-like (C-M) or thin (T-M) without branching. The growth rate of the transformed root lines was always higher than that of untransformed roots, showing that the genetic changes caused by the A. rhizogenes transformation conditioned a higher biomass formation. When considering the different transformed root phenotypes, we can observe that the highest ginsenoside production was achieved by HR-M root lines, closely followed by C-M ones, whereas the lowest yield was reached by T-M root phenotype. The study of the integration of the TL-DNA and TR-DNA fragments of the pRiA4 in the root genome showed that the aux1 gene was always detected in HR-M and C-M root phenotypes which presented the highest biomass and ginsenoside productions. This fact suggests a significant role of aux genes in the morphology of Panax ginseng transformed roots. The ginsenoside pattern of transformed roots varied according to their morphology, although the ginsenoside contents of the Rg group was always higher than that of the Rb group. From our results, we can infer the potential of some root phenotypes of Panax ginseng hairy root cultures for an improved ginsenoside production. PMID- 11393516 TI - Studies on the biosynthesis of 2-hydroxy-1,4-benzoxazin-3-one (HBOA) from 3 hydroxyindolin-2-one in Zea mays. AB - The ring expansion of 3-hydroxyindolin-2-one to 2-hydroxy-1,4-benzoxazin-3-one (HBOA) was investigated by labelling experiments. Action of the cytochrome P450 enzyme BX4 from maize on 3-hydroxyindolin-2-one under an 18O2 atmosphere induced production of 2-hydroxy-1,4-benzoxazin-3-one in which the ring oxygen--but not the 2-hydroxy group of HBOA--is labelled. A mechanism for this transformation is proposed. PMID- 11393517 TI - Biotransformation of squamulosone by Curvularia lunata ATCC 12017. AB - Squamulosone (aromadendr-1(10)-en-9-one), isolated in large quantity from the plant Hyptis verticillata Jacq. (Labiatae), was incubated with the fungus Curvularia lunata ATCC 12017 in two different growth media. Six metabolites were isolated from each medium. with five of the products being common to both fermentations. All seven metabolites are novel. The insecticidal activity of these aromadendranes was evaluated against the sweet potato weevil Cylas formicarius elegantus. PMID- 11393518 TI - Blend effects in the toxicity of the essential oil constituents of Ocimum kilimandscharicum and Ocimum kenyense (Labiateae) on two post-harvest insect pests. AB - The lethal toxicity of major components of the essential oils of Ocimium kilimandscharicum and O. kenyense and of selected blends of these against Sitophilus zeamais and Rhyzopertha dominica were compared with those of the full blends of the essential oils. The compounds were assayed in amounts and proportions present in the minimum 100% lethal dose of the oils. Whereas a major component of O. kilimandscharicum was found to be largely responsible for the toxic action of its essential oil against R. dominica, the results with the other treatments indicated that the toxic action of the essential oils were due to the combined effects of different components, either with or without significant individual toxic action of their own against the insects. The significance of the results and their implication in screening and using plants and their phytochemicals for pest and microbial control are highlighted. PMID- 11393519 TI - Identification and ecdysteroid antagonist activity of three oligostilbenes from the seeds of Carex pendula (Cyperaceae). AB - Methanolic extracts of seeds of several (Carex species were found to antagonise the action of 20-hydroxyecdysone in the Drosophila melanogaster microplate-based B(II) cell bioassay. Bioassay-guided HPLC analysis of seeds of Carex pendula (drooping sedge) provided one previously unknown tetrastilbene (cis-miyabenol A) and two known oligostilbenes (kobophenol B and cis-miyabenol C) as the biologically active compounds (EC50 values were 31, 37 and 19 microM, respectively, vs. 5 x 10(-8) M 20-hydroxyecdysone). The structures and relative stereochemistries of these compounds were deduced by comprehensive ID- and 2D-NMR experiments. These compounds are isolated from Carex pendula for the first time. In vitro experiments with dipteran and lepidopteran ecdysteroid receptor proteins demonstrate that the oligostilbenes are able to compete with radiolabelled ecdysteroid ([3H]ponasterone A) for occupancy of the ligand binding site. IC50/Ki values are similar to the EC50 values obtained in the B(II) bioassay. PMID- 11393520 TI - Ecdysteroids and bufadienolides from Helleborus torquatus (Ranunculaceae). AB - Three bufadienolides, hellebortin A (5-[beta-D-glucopyranosyloxy]-10,14,16 trihydroxy-19-nor-[5beta,10beta,14beta,16beta]-bufa-3,20,22-trienolide [1]), hellebortin B (5-[beta-D-glucopyranosyloxy]-3,4-epoxy-14-hydroxy-19-oxo-bufa 20,22-dienolide [2]) and hellebortin C (5-[beta-D-glucopyranosyloxy]-3,4-epoxy 10,14-dihydroxy-19-nor-bufa-20,22-dienolide [3]), together with 20 hydroxyecdysone 3-O-beta-D-glucoside (4) and 20-hydroxyecdysone (5) have been isolated by bioassay- and RIA-directed HPLC analyses of a methanol extract of the seeds of Helleborus torquatus. The structure and relative stereochemistry of the novel bufadienolide hellebortin A (1) and the structures of hellebortin B (2) and hellebortin C (3) were determined unambiguously by comprehensive analyses of their 1D and 2D NMR data. These five compounds are isolated from Hellborus torquatus for the first time. The biological activities of compound 1, 4 and 5 as ecdysteroid agonists and antagonists have been assessed. PMID- 11393521 TI - Induced emissions of apple fruit volatiles by the codling moth: changing patterns with different time periods after infestation and different larval instars. AB - The changes in the emission of volatiles from mature apple fruits in response to larval feeding by the codling moth (Cydia pomonella) under laboratory conditions are reported. A time course experiment investigated the emission of volatiles throughout the period of larval development following infestation. The volatiles consisted mainly of esters, a few aldehydes, and the terpene alpha-farnesene. Infested apples emitted the same compounds as healthy apples. The quantities of volatiles released were much higher for infested as compared to healthy fruits for an initial three day period. Following this period there was a decrease in volatile emissions (days 6-9), eventually declining back to the levels emitted from healthy apples or below by 9-21 days after infestation. In a separate experiment, the volatile emissions from healthy and artificially damaged fruits were compared to those from herbivore damaged fruits for each of the five larval instars of C. pommonella. The results from the discriminant analysis indicate that the most effective induction of volatiles occurred when fruits were infested with first instar larvae. Induction by first instar larvae was generally higher than after infestation by later instars, and for most compounds it also exceeded the emission from artificially damaged fruits. PMID- 11393522 TI - Methyl jasmonate increases reported alkamides and ketoalkene/ynes in Echinacea pallida (Asteraceae). AB - Methyl jasmonate (MJ), a naturally-occurring plant cellular signal molecule, was found to induce production of lipophilic secondary metabolites in Echinacea pallida seedlings. Seedling aerial parts were sprayed with 100 ppm MJ, and roots were harvested and extracted 24 h later. Lipophilic root extracts, separated by HPLC, revealed significant increases (P< 0.05) in six alkamides or related ketoalkene/ynes produced by 34 day-old plants and in seven compounds produced by 58 day-old plants. PMID- 11393523 TI - Antiplatelet activity of N-methoxycarbonyl aporphines from Rollinia mucosa. AB - Bioassay-directed fractionation of the stems of Rollinia mucosa led to the isolation of new N-methoxycarbonyl aporphine alkaloids, romucosine A (1), romucosine B (2), romucosine C (3), and romucosine D (4), along with the known alkaloid, N-methoxylcarbonyl-nornuciferine (5). Alkaloids 1 and 4 exhibited significant inhibition of collagen, arachidonic acid, and platelet activating factor-induced platelet aggregation, and alkaloid 3 also showed an inhibitory effect on arachidonic acid induced platelet aggregation. PMID- 11393524 TI - Benzyl isothiocyanate is the chief or sole anthelmintic in papaya seed extracts. AB - Papaya (Carica papaya) seeds were extracted in an aqueous buffer or in organic solvents, fractionated by chromatography on silica and aliquots tested for anthelmintic activity by viability assays using Caenorhabditis elegans. For all preparations and fractions tested, anthelmintic activity and benzyl isothiocyanate content correlated positively. Aqueous extracts prepared from heat treated seeds had no anthelmintic activity or benzyl isothiocyanate content although both appeared when these extracts were incubated with a myrosinase containing fraction prepared from papaya seeds. A 10 h incubation of crude seed extracts at room temperature led to a decrease in anthelmintic activity and fractionated samples showed a lower benzyl isothiocyanate content relative to non incubated controls. Benzyl thiocyanate, benzyl cyanide, and benzonitrile were not detected in any preparations and cyanogenic glucosides. which were present, could not account for the anthelmintic activity detected. Thus, our results are best explained if benzyl isothiocyanate is the predominant or sole anthelmintic agent in papaya seed extracts regardless of how seeds are extracted. PMID- 11393525 TI - Lipophyllic antioxidants from Iryanthera juruensis fruits. AB - The phytochemical investigation of the hexane extract of Iryanthera juruensis (Myristicaceae) fruits led to the isolation of two tocotrienols and four lignans which exhibited antioxidant activity towards beta-carotene on TLC autographic assay. Two inactive quinones and three omega-arylalkanoic acids were also isolated. The isolates were investigated for their redox properties using cyclic voltammetry. The structure elucidation of the new compounds (one tocotrienol. one quinone and three omega-arylalkanoic acids) was based on analysis of spectroscopic data. PMID- 11393526 TI - Volatile constituents in mosses (Musci). AB - The essential oils of mosses of the genera Mnium, Plagiomnium, Homalia, Plagiothecium and Taxiphyllum (Musci) have been investigated by gas chromatography and mass spectrometry. The new sesquiterpenes (+)-10-epi-muurola 4,11-diene and 10,11-dihxdro-alpha-cuparenone were isolated by preparative gas chromatography and identified as major constituents of the hydrodistillation products of Mnium hornmum (Hedw.) using NMR and mass spectrometry. In addition, (+)-dauca-8,11-diene and two new butenolides, 3,4,5-trimethyl-5-pentyl-5H-furan-2 one and 3.4-dimethyl-5-pentyl-5H-furan-2-one were identified as constituents in Plagiomnium undulatum (Hedw.) T. Kop. Although the amounts of volatiles present in the investigated mosses are generally smaller than in liverworts, the spectrum of terpenoid compounds is similar. The investigated mosses also generate aliphatic compounds of greater abundance and structural variety. PMID- 11393527 TI - High molecular weight lipids from the trilaminar outer wall (TLS)-containing microalgae Chlorella emersonii, Scenedesmus conmmunis and Tetraedron minimum. AB - High molecular weight lipids were isolated from Chlorella emersonii, Scenedesmus communis and Tetraedron minimum, thin trilaminar outer wall (TLS)-containing freshwater microalgae producing an insoluble non-hydrolysable biopolymer (i.e. algaenan). Molecular weight determination by gel permeation chromatography indicated that their molecular weights range from ca. 400 to 2000 Da. Flash pyrolysis with in situ methylation using tetramethylammonium hydroxide (TMAH) and alkaline hydrolysis showed that the high molecular weight lipids isolated from C. emersonii and S. communis are mainly composed of saturated n-C26 and n-C28 fatty acids and alcohols and of saturated n-C30 and n-C32 alpha,omega-diols and omega hydroxy acids. In contrast the high molecular weight lipids isolated from T. minimum are predominantly composed of long-chain fatty acids and omega-hydroxy acids. Aromatic moieties were also identified in small amounts in the thermochemolysate and in the hydrolysate. Chemical structural models containing long-chain mono- and polyesters were proposed for the high molecular weight lipids isolated from the three microalgae in agreement with analytical and spectroscopic data. Structural similarity between the outer cell wall of these microalgae and the cuticular membrane of higher plants is suggested. PMID- 11393528 TI - Triterpenoid saponins from the stem bark of Elattostachys apetala. AB - Six new triterpenoid saponins have been isolated from the stem bark of Elattostachys apetala together with four known triterpenoid saponins. Three of these new compounds are glycosides of a newly described genin, 29 hydroxyhederagenin (1). On the basis of spectral evidence, the structures of the new saponins were concluded to be alpha-hederin 28-O-[alpha-L-rhamnopyranosyl-(1- >2)-beta-D-glucopyranosyl] ester (2), sapindoside B 28-O-[alpha-L-rhamnopyranosyl (1-->2)-beta-D-glucopyranosyl] ester (3), 3-O-beta-D-xylopyranosyl astrantiasaponin VII (4), 3-O-[alpha-L-rhamnopyranosyl-(1-->2)-alpha-L arabinopyranosyl]-28-O-[beta-D-glucopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1- >6)]-beta-D-glucopyranosyl]-29-hydroxyhederagenin (5), 3-O-[alpha-L arabinopyranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L arabinopyranosyl]-28-O-[beta-D-glucopyranosyl(1-->2)-[beta-D-glucopyranosyl(1- >6)]-beta-D-glucopyranosyl]-29-hydroxyhederagenin (6), and 3-O-[beta-D xylopyranosyl-(1-->3)-alpha-L-rhamnopyranosyl-(1-->2)-alpha-L-arabinopyranosyl] 28-O-[beta-D-glucopyranosyl-(1-->2)-beta-D-gluco pyranosyl-(1-->6)]-beta-D glucopyranosyl]-29-hydroxyhederagenin (7). PMID- 11393529 TI - Hydroquinone derivatives and monoterpenoids from the Neotropical liverwort Plagiochila rutilans. AB - The structure of a prenylbenzene derivative isolated previously from a Cuban specimen of the liverwort Plagiochila rutilans is revised to 2-methoxy-6 prenylhydroquinone. The hydroquinone was observed as a prominent component of the NMR and GC-MS fingerprints of five recent specimens of the liverwort from Bolivia, Brazil and Costa Rica. The corresponding quinone was observed as a minor component. Two new methylated derivatives of the hydroquinone were observed as prominent components in one specimen from Bolivia; these were isolated, characterized, and their structures elucidated as 2-methoxy-1-O-methyl-6 prenylhydroquinone and 2-methoxy-4-O-methyl-6-prenylhydroquinone using 1H NMR spectroscopy. The liverwort has a strong peppermint-like odour that is caused by the presence of several menthane monoterpenoids, including notably pulegone, menthone, isomenthone. terpinolene and limonene. One of the Costa Rican specimens contained considerable amounts of the new lactone 3,7-dimethyl-2,6-octadien-1,6 olide as the principal monoterpenoid in place of pulegone. Two Costa Rican specimens distinguished morphologically as Plagiochila standleyi (a taxon closely related to P. rutilans and reduced elsewhere to a variety of that species) are characterized by large amounts of 3-hydroxy-4'-methoxybibenzyl. P. standleyi was also reported to have a peppermint-like odour in the field. Menthane monoterpenoids were again responsible but in this case the major components were limonene, beta-phellandrene, alpha-terpinene and the endoperoxide ascaridole. PMID- 11393530 TI - Application of MS and NMR to the structure elucidation of complex sugar moieties of natural products: exemplified by the steroidal saponin from Yucca filamentosa L. AB - An approach, using well characterized procedures, is presented that should be of general applicability for the structural elucidation of complex sugar moieties of natural products. The methods used are exemplified by the structure elucidation of a new gitogenin-based steroidal saponin that has a strong leishmanicidal activity similar to preparations used in clinical practice and has been isolated by bioactivity-guided fractionation of the ethanolic extract of Yucca filamentosa L. leaves. The saponin has been characterized as 3-O-((beta-D-glucopyranosyl-(1- >3)- beta-D-glucopyranosy-(1-->2))(alpha-L-rhamnopyranosyl-(1-->4)-beta-D glucopyranosyl-(1-->3))-beta-D-glucopyranosyl-(1-->4)-beta-D-galactopyranosyl) 25R,5alpha-spirostan-2alpha,3beta-diol. PMID- 11393531 TI - Development of genetic hypotheses in essential hypertension. AB - Essential hypertension illustrates the formidable task presented by the identification of genetic determinants of common disease. Making an initial genetic inference may prove difficult enough; the subsequent demonstration of functional significance at various levels of biological integration may be even more challenging. We review three instances in which an initial genetic inference has led to the development of testable hypotheses pursued at increasingly higher levels of biological organization. These include the adducin, the G protein beta3 subunit, and the angiotensinogen hypotheses. PMID- 11393532 TI - Phenotypes of X-linked Charcot-Marie-Tooth disease and altered trafficking of mutant connexin 32 (GJB1). AB - To clarify the pathomechanism in three patients with X-linked Charcot-Marie-Tooth disease (CMTX) and unique clinical features, we studied three connexin (Cx) 32 (GJB1) mutants with respect to cellular localization in cultured cells. Wild-type Cx32 and three Cx32 mutants (Va163Ile and Glu186Lys, obtained from CMTX patients with hearing impairment; and Arg22Gln, obtained from a CMTX patient with a fair number of onion-bulb formations) were transfected to rat pheochromocytoma cells (PC12). We investigated the expression of Cx32 protein in each clone by immunoblotting and immunohistochemical staining. While Cx32 protein with the Arg22Gln mutation was detectable immunohistochemically only in the cytoplasm, Cx32 protein with the Va163Ile or Glu186Lys mutation was detected in both the plasma membrane and the cytoplasm. Cx32 protein with the wild-type sequence was detected mostly in the plasma membrane, with plaques indicating the existence of active gap junction formation. These three Cx32 mutations associated with CMTX patients with unique clinical and pathological findings caused altered trafficking of the Cx32 protein. These altered expressions indicated loss of active gap junction formation with different expression abnormalities in these CMTX patients. PMID- 11393533 TI - Identification of single-nucleotide polymorphisms (SNPs) of human N acetyltransferase genes NAT1, NAT2, AANAT, ARD1 and L1CAM in the Japanese population. AB - By direct sequencing of regions of the human genome containing five genes belonging to the acetyltransferase family, arylamine N-acetyltransferase (NAT1), arylamine N-acetyltransferase (NAT2), arylalkylamine N-acetyltransferase (AANAT), L1 cell adhesion molecule (L1CAM), and the human homolog of Saccharomyces cerevisiae N-acetyltransferase ARD1, we identified 53 single-nucleotide polymorphisms (SNPs) and two insertion/ deletion polymorphisms in 48 healthy Japanese volunteers. NAT1 and NAT2 are so-called drug-metabolizing enzymes. In the NAT1 gene we found two SNPs and a 3-bp insertion/ deletion polymorphism that corresponded to the NAT1*3, *10, and *18A/*18B alleles reported in other populations. The frequencies of NAT1* alleles in our Japanese subjects were 52.6% for NAT1*4, 1.0% for NAT1*3, 40.6% for NAT1*10, 2.6% for NAT1*18A and 3.1% for NAT1*18B. In the NAT2 gene we found 32 SNPs and a 1-bp insertion/ deletion polymorphism; 6 SNPs within the coding region were reported previously and belonged to the slow acetylator group (NAT2*5, NAT2*6 and NAT2*7), and 2 of the 8 SNPs in the 5' flanking region were reported in the dbSNP of GenBank, but the remaining 24 SNPs and the insertion/deletion polymorphism were novel. The frequencies of NAT2* alleles in Japanese (51.3% for NAT2*4, 1.6% for *5B, 26.1% for *6A, 2.2% for *6B, 1.2% for *7A, 10.1% for *7B, 7.4% for *12A, and 1.1% for *13) were significantly different from those reported in Caucasian populations. In the AANAT gene we found 4 novel SNPs: 2 in the 5' flanking region, 1 in exon 4, and 1 in intron 3. In the two genes belonging to the N-terminal N acetyltransferase family, we identified 9 SNPs, 7 of them novel, for ARD1, and six novel SNPs for L1CAM. Variations at these loci may contribute to an understanding of the way in which different genotypes may affect the activities of human N-acetyltransferases, especially as regards the therapeutic efficacy of certain drugs and antibiotics. PMID- 11393534 TI - Polymorphisms in PNLIP, encoding pancreatic lipase, and associations with metabolic traits. AB - Pancreatic lipase (EC 3.1.1.3) is an exocrine secretion that hydrolyzes dietary triglycerides in the small intestine. We developed genomic amplification primers to sequence the 13 exons of PNLIP, which encodes pancreatic lipase, in order to screen for possible mutations in cell lines of four children with pancreatic lipase deficiency (OMIM 246600). We found no missense or nonsense mutations in these samples, but we found three silent single-nucleotide polymorphisms (SNPs), namely, 96A/C in exon 3, 486C/T in exon 6, and 1359C/T in exon 13. In 50 normolipidemic Caucasians, the PNLIP 96C and 486T alleles had frequencies of 0.083 and 0.150, respectively. The PNLIP 1359T allele was absent from Caucasian, Chinese, South Asian, and North American aboriginal samples, but had a frequency of 0.085 in an African sample, suggesting that it is a population-specific variant. In an association analysis of 185 African neonates, the PNLIP 1359C/T SNP genotype was significantly associated with concentrations of plasma lipoproteins. These associations were most likely due to linkage disequilibrium with another functional variant at or near PNLIP. Thus, we report three new SNPs for the PNLIP, which may serve as markers for association analyses and for pharmacogenetic studies of pancreatic lipase inhibitors. PMID- 11393535 TI - Seventy genetic variations in human microsomal and soluble epoxide hydrolase genes (EPHX1 and EPHX2) in the Japanese population. AB - Human microsomal and soluble epoxide hydrolases (mEH and sEH) are enzymes that metabolize xenobiotic molecules. We screened DNA from 48 Japanese individuals for single-nucleotide polymorphisms (SNPs) in both genes by direct sequencing of the entire genomic regions containing EPHX1 and EPHX2, except for repetitive elements. This approach identified 33 SNPs in the EPHX1 gene; 6 of them were located in the 5' flanking region, 17 in introns, 8 in exons, and 2 in the 3' flanking region. In the EPHX2 gene, we identified 36 SNPs, including 4 in the 5' flanking region, 24 in introns, 5 in exons, and 3 in the 3' flanking region, as well as one insertion/deletion polymorphism in the 5' flanking region. These variants may contribute to a more precise understanding of the nature of correlations between genotypes and disease-susceptibility phenotypes that have been postulated in regard to human microsomal and soluble epoxide hydrolases. PMID- 11393536 TI - Prevalence of A-to-G mutation at nucleotide 3243 of the mitochondrial tRNA(Leu(UUR)) gene in Japanese patients with diabetes mellitus and end stage renal disease. AB - The A-to-G mutation at nucleotide 3243 of the mitochondrial tRNA(Leu(UUR)) gene (mt.3243A>G) is associated with both diabetes mellitus and myopathy, encephalopathy, lactic acidosis, and stroke-like episodes (MELAS). Recently, this mutation was found in three diabetic subjects with progressive kidney disease, suggesting that it may be a contributing factor in the development of kidney disease in patients with diabetes. The aim of this study was to evaluate the contribution of this mutation to the development of end stage renal disease (ESRD) in patients with diabetes. The study group consisted of 135 patients with diabetes and ESRD. The control group consisted of 92 non-diabetic subjects with ESRD who were receiving hemodialysis. The mt.3243A>G mutation was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). We found the mt.3243A>G mutation in eight patients (8/135; 5.9%), all of whom were initially diagnosed with type II diabetes. Five of the eight patients were subsequently also diagnosed with MELAS. We did not find the mutation in any of the 92 nondiabetic subjects with ESRD. The prevalence of this mutation was 6.5 fold higher in patients with diabetes and ESRD than in those with diabetes alone (8/135 vs 5/550, respectively; chi2 = 13.704; P = 0.0002). The mt.3243A>G mutation may be a contributing genetic factor in the development of ESRD in Japanese patients with diabetes. PMID- 11393537 TI - Frequent loss of heterozygosity at 3p25-p26 is associated with invasive oral squamous cell carcinoma. AB - Recent molecular evidence suggests that allelic deletions of chromosomes are involved in the carcinogenesis of various neoplasms, including oral squamous cell carcinoma (OSCC). To determine the role of 3p deletions in Japanese OSCC and to define the localization of putative tumor suppressor genes, we initially examined loss of heterozygosity (LOH), using nine microsatellite markers in 36 OSCCs and 28 oral epithelial dysplastic lesions (OEDLs). LOH on chromosome 3p was observed at one or more loci in 72% of OSCCs and 18% of OEDLs. Fourteen (61%) of 23 OSCC patients informative at D3S2450 (3pter-p24.2) showed LOH most frequently, in contrast to OEDL, where LOH was never seen at this locus. Interestingly, we found a significant association between an allelic deletion at this locus and the histologic grade of mode of tumor invasion. Therefore, we also examined allelic deletion on chromosome 3p telomeric to where D3S2450 was located. A common deletion region was identified between D3S2450 and D3S3591. Our results provide evidence for the presence of a tumor suppressor gene in a 0.8-cM region bordered by D3S2450 and D3S3591 at 3p25-p26, which may play a role in carcinogenesis and invasion of OSCC. PMID- 11393538 TI - Human arylhydrocarbon receptor repressor (AHRR) gene: genomic structure and analysis of polymorphism in endometriosis. AB - The diversity of biological effects resulting from exposure to dioxin may reflect the ability of this environmental pollutant to alter gene expression by binding to the arylhydrocarbon receptor (AHR) gene and related genes. AHR function may be regulated by structural variations in AHR itself, in the AHR repressor (AHRR), in the AHR nuclear translocator (ARNT), or in AHR target molecules such as cytochrome P-4501A1 (CYP1A1) and glutathione S-transferase. Analysis of the genomic organization of AHRR revealed an open reading frame consisting of a 2094 bp mRNA encoded by ten exons. We found one novel polymorphism, a substitution of Ala by Pro at codon 185 (GCC to CCC), in exon 5 of the AHRR gene; among 108 healthy unrelated Japanese women, genotypes Ala/Ala, Ala/Pro, and Pro/Pro were represented, respectively, by 20 (18.5%), 49 (45.4%), and 39 (36.1%) individuals. We did not detect previously published polymorphisms of ARNT (D511N) or the CYP1A1 promoter (G-469A and C-459T) in our subjects, suggesting that these polymorphisms are rare in the Japanese population. No association was found between uterine endometriosis and any polymorphisms in the AHRR, AHR, ARNT, or CYP1A1 genes analyzed in the present study. PMID- 11393539 TI - Parental age and the origin of extra chromosome 21 in Down syndrome. AB - We present a report of the parental ages (n = 865) and parental origin of meiotic nondisjunction (n = 236) that are likely to show a predisposition in the etiology of Down syndrome (DS). Chromosomal analysis, performed over a 20-year period, on 1,001 Down syndrome subjects, revealed pure trisomy 21 karyotype in 880 subjects (87.92%), mosaic trisomy karyotype in 77 (7.69%), and translocation karyotype in 44 (4.39%). The mean maternal age was found to be 30.34 years, and mean paternal age was 31.04 years. Nondisjunctional error was 79.24% maternal and 20.76% paternal. The findings of the study revealed the significant contribution of advanced parental age and increased maternal meiotic nondisjunctional error to the origin of trisomy 21 Down syndrome. PMID- 11393540 TI - Single-nucleotide polymorphisms of the nuclear lamina proteome. AB - Familial partial lipodystrophy (FPLD) has been shown to be due to mutations in the LMNA gene encoding nuclear lamins A and C, indicating that defective structure of the nuclear envelope can produce this unique phenotype. Some patients with inherited partial lipodystrophy have normal LMNA coding, promoter, and 3'-untranslated region sequences. This suggests that the FPLD phenotype is genetically heterogeneous. Among the candidate genes to consider for the non-LMNA associated forms of FPLD are other components of the inner nuclear membrane, such as lamin B1 and B2 and the lamin B receptor. We developed amplification primers for the coding regions of LMNB1, LMNB2, and LBR, which encode lamin B1, lamin B2, and the lamin B receptor, respectively. We found no putative disease mutations in any of these proteins in subjects with non-LMNA FPLD, but, through the screening of diseased and normal subjects, we identified several single-nucleotide polymorphisms (SNPs); specifically, five SNPs in LMNB1 and four SNPs in LBR. The LMNB2 gene was monomorphic in screening experiments. We conclude that mutations in other constituent proteins of the nuclear envelope are not present in subjects with non-LMNA-associated FPLD. However, the identification of amplification primers and SNPs provides tools to investigate these proteins for their association with other phenotypes. PMID- 11393541 TI - PM2.5 and PM10 concentrations from the Qalabotjha low-smoke fuels macro-scale experiment in South Africa. AB - This article presents results from the particulate monitoring campaign conducted at Qalabotjha in South Africa during the winter of 1997. Combustion of D-grade domestic coal and low-smoke fuels were compared in a residential neighborhood to evaluate the extent of air quality improvement by switching household cooking and heating fuels. Comparisons are drawn between the gravimetric results from the two types of filter substrates (Teflon-membrane and quartz-fiber) as well as between the integrated and continuous samplers. It is demonstrated that the quartz-fiber filters reported 5 to 10% greater particulate mass than the Teflon-membrane filters, mainly due to the adsorption of organic gases onto the quartz-fiber filters. Due to heating of sampling stream to 50 degrees C in the TEOM continuous sampler and the high volatile content of the samples, approximately 15% of the particulate mass was lost during sampling. The USEPA 24-hr PM2.5 and PM10 National Ambient Air Quality Standards (NAAQS) of 65 microg m(-3) and 150 microg m(-3), respectively, were exceeded on several occasions during the 30-day field campaign. Average PM concentrations are highest when D-grade domestic coal was used, and lowest between day 11 and day 20 of the experiment when a majority of the low-smoke fuels were phased in. Source impacts from residential coal combustion are also found to be influenced by changes in meteorology, especially wind velocity. PM2.5 and PM10 mass, elements, water-soluble cations (sodium, potassium, and ammonium), anions (chloride, nitrate, and sulfate), as well as organic and elemental carbon were measured on 15 selected days during the field campaign. PM2.5 constituted more than 85% of PM10 at three Qalabotjha residential sites, and more than 70% of PM10 at the gradient site in the adjacent community of Villiers. Carbonaceous aerosol is by far the most abundant component, accounting for more than half of PM mass at the three Qalabotjha sites, and for more than a third of PM mass at the gradient site. Secondary aerosols such as sulfate, nitrate, and ammonium are also significant, constituting 8 to 12% of PM mass at the three Qalabotjha sites and 15 to 20% at the Villiers gradient site. PMID- 11393542 TI - Water quality assessment of Osun River: studies on inorganic nutrients. AB - The present investigation provides data of some ions, namely Na+, Ca2+, NH4+, Cl , NO3-, CN- and PO4(3-) on water samples of river Osun, selected rivers in the region and groundwaters. The pH, temperature, electrical conductivity (EC), total dissolved solids (TDS), total hardness (TH) and total carbon (IV) oxide (TCO2) have also been determined to asses the chemical status and pollution levels of these water sources. The higher values of certain parameters with respect to the acceptable standard limits for drinking water indicate the pollution in both groundwater and river water samples of the study area, and make the waters unsuitable for various applications. The high pollution river water source showed higher levels of phosphate, nitrate and ammonium ions (P < 0.05). There is no significant difference (P < 0.05) between the mean concentrations of other inorganic nutrients in the high and low pollution water source types. The correlation coefficient between quality parameter pairs of river water and groundwater samples are determined and the significance of these parameters in both types of water sources are discussed. PMID- 11393543 TI - Duck populations as indicators of landscape condition in the Prairie Pothole Region. AB - The Prairie Pothole Region of the northern Great Plains is an important region for waterfowl production because of the abundance of shallow wetlands. The ecological significance of the region and impacts from intensive agriculture prompted the U.S. Environmental Protection Agency to select it as one of the first areas for developing and evaluating ecological indicators of wetland condition. We examined hypothesized relations between indicators of landscape and wetland conditions and waterfowl abundance on 45 40 km2 study sites in North Dakota for 1995-1996. Landscape condition was defined a priori as the ratio of cropland area to total upland area surrounding wetlands. Measures of waterfowl abundance included estimated numbers of breeding pairs (by species and total numbers) and gamma, a species-specific correction factor which effectively adjusts breeding pair estimates for annual or area-related differences in pond size. Landscape indicators and waterfowl measures varied among regions. Results indicated that most areas in the Coteau region are of much higher quality for ducks than those in the Drift Plain, and areas in the Red River Valley are of the poorest quality for ducks. Regression models demonstrated the impact of agricultural development on breeding duck populations in the Prairie Pothole Region. The most consistent landscape indicators of waterfowl abundance were percent of cropland and grassland. Models were inconsistent among years and species. The potential biotic indicators of landscape and wetland condition examined here would be appropriate for temporal trend analyses, but because of inherent geographic variability would not be appropriate for single-year geographic trend analyses without more extensive evaluations to improve explanatory models. PMID- 11393544 TI - Beryllium concentrations in ambient air and its source identification. A case study. AB - Beryllium concentrations in atmospheric particulate and soil samples in and around a Beryllium Processing Facility (BPF) have been measured. The mean air concentration level of beryllium in and around the fence line of the BPF is 0.48 +/- 0.43 ng m(-3) (n = 397) and is mostly influenced by diurnal and seasonal changes. The observed air concentration levels were well below the prescribed ambient air quality (AAQ) standard of 10 ng m(-3). The soil concentration levels of beryllium in the study area were found to be in the range of 1.42-2.75 microg g(-1). The mass median aerodynamic diameter (MMAD) of beryllium aerosols in ambient air was found to be 6.9 microm. Source identification using the Enrichment Factor (EF) approach indicates soil as the predominant contributory source for air concentrations at the site. PMID- 11393545 TI - Stream phosphorus transport in the Lake Tahoe basin, 1989-1996. AB - Lake Tahoe is undergoing the initial stages of cultural eutrophication due to human alteration of the airshed and watershed. The lake's switch from nitrogen (N) to phosphorus (P) limitation has been attributed primarily to atmospheric N loading. This places an increased importance on controlling watershed movement of P to the lake. A stream water quality monitoring data set consisting of nine streams in the Lake Tahoe basin has been analyzed to characterize the spatiotemporal variation of P delivery to the lake. This data is from the Lake Tahoe Interagency Monitoring Program (LTIMP), which provides scientific data for planning and regulatory agencies to address environmental problems in the Lake Tahoe basin. Results indicate that P delivery (concentrations, loads) varies greatly at interannual, seasonal, and spatial scales. Annual and seasonal total P (TP) concentrations can vary up to three orders of magnitude in a given stream and are strongly associated with suspended sediment. Particulate P is the major form of P transported by Tahoe streams and was strongly correlated with percent surficial geologic deposits, which are primarily located near streams. Tahoe streams with the highest annual P concentrations often had the lowest annual P loads, and visa versa. P loading is greatest during the spring snowmelt (75% of annual average). Potential watershed parameters influencing P delivery to Lake Tahoe have been identified as precipitation, basin area, basin steepness, and road and human development coverage. Results also suggest that human development impacts on stream P loads are most prevalent during high precipitation years. Identification and quantification of stream sediment and P sources such as streambanks and impervious surface is necessary to aid in watershed restoration efforts. PMID- 11393546 TI - Statistical analysis of spatial pattern: a comparison of grid and hierarchical sampling approaches. AB - Previous studies have combined random-site hierarchical sampling designs with analysis of variance techniques, and grid sampling with spatial autocorrelation analysis. We illustrate that analysis techniques and sampling designs are interchangeable using densities of an infaunal bivalve from a study in Poverty Bay, New Zealand. Hierarchical designs allow the estimation of variances associated with each level, but high-level factors are imprecisely estimated, and they are inefficient for describing spatial pattern. Grid designs are efficient for describing spatial pattern, and are amenable to conventional analysis. Our example deals with a continuous spatial habitat, but our conclusions also apply in disjunct or patchy habitats. The influence of errors in positioning is also assessed. The advantages of systematic sampling are reviewed, and more efficient hierarchical approaches are identified. The distinction between biological and statistical significance in all analyses is emphasised. PMID- 11393547 TI - The history and impact of the International Beilinson Symposia. PMID- 11393548 TI - Disease-associated autoantibodies as predictive markers of type 1 diabetes mellitus in siblings of affected children. AB - The long latent preclinical period of type 1 diabetes mellitus (DM) makes it possible to identify individuals at increased risk for clinical DM before the beta-cell destructive process has reached the point of no return. A series of reports on the predictive value of DM-associated autoantibodies are available in first-degree relatives of patients with type 1 DM, but only a few of these studies target exclusively siblings and young siblings of affected children. When planning screening of siblings for DM risk, their age needs to be considered, as predictive characteristics of autoantibodies seem to vary in different age groups. Autoimmunity may be initiated early in life and therefore early screening for signs of beta-cell autoimmunity is crucial to avoid missing young children en route to overt DM and to be able to start intervention, when clinically applicable preventive modalities become available, before the disease process has advanced too far. Young age, positivity for at least two autoantibodies, high levels of autoantibodies and low first phase insulin response are highly predictive for progression to clinical disease in initially unaffected siblings of children with type 1 DM. PMID- 11393549 TI - Familial predisposition of type 1 diabetes mellitus in Japan, a country with low incidence. Japan Diabetes Society Data Committee for Childhood Diabetes. AB - The study group of the Japan Diabetes Society (JDS) carried out nationwide hospital-based and population-based surveys of childhood type 1 diabetes mellitus (DM) in Japan. According to the nationwide population-based survey, the incidence of childhood type 1 DM in Japan was 1.5 (1.4-1.6)/10(5), which did not differ for the 5 years from 1986-1990. Predisposition for DM and autoimmunity were studied in the first-degree relatives of the patients, including older and later cohorts. The prevalence of type 1 DM was 3.3% (12/369) among siblings and 2.2% (8/369) among parents, while the prevalence of type 2 DM was 0% among siblings and 4.3% (16/369) among parents. The risk of type 1 DM among siblings of the patients was 330 times higher than that among the general population in the Japanese population. The rate of positivity for autoantibodies, including ICA, IAA, GAD and IA-2, was 1.4-2.9% in parents (n=140) and 2.0-3.9% in siblings (n=203). The genetic susceptibility for type 1 DM is far lower in Japanese children than in Caucasian children, but predisposition to the disease and positive autoimmunity are almost the same in Japanese families of patients as in Caucasian families. The quality of life of Japanese parents of children with type 1 DM was less satisfactory that that of the Caucasian parents previously reported, which might be a result of the low incidence of type 1 DM in Japan. PMID- 11393550 TI - Celiac disease and its link to type 1 diabetes mellitus. AB - Celiac disease (CD) is a small intestinal disorder with overt malabsorption in the minority and with subclinical or atypical symptoms in the majority of patients. It is triggered by gluten and related cereal proteins in a unique genetic background (HLA-DQ2 or DQ8 and other unmapped genes). CD is characterized by a highly specific mucosal autoantibody response to tissue transglutaminase. In the intestine this enzyme creates antigenic neoepitopes in gluten peptides which are more efficiently presented to the immune system in the context of HLA-DQ2 or DQ8. Between 3% and 6% of patients with type 1 diabetes mellitus (DM) have (atypical) CD, and the prevalence of a variety of autoimmune diseases in patients with CD correlates with the time of gluten exposure, reaching 35% after 20 years. It is still unknown whether oligosymptomatic CD favors the development of type 1 DM and whether a gluten-free diet modifies the progression of DM in general. Apart from shared or adjacent HLA loci in both diseases, post-translational modification of potential autoantigens by enzymes such as tissue transglutaminase could play a role in the autoimmunity of type 1 DM. PMID- 11393551 TI - Type 1 diabetes mellitus, celiac disease and their association--lessons from antibodies. PMID- 11393552 TI - Diagnosis of MODY in the offspring of parents with insulin-dependent and non insulin-dependent diabetes mellitus. AB - Maturity Onset Diabetes of the Young (MODY) is an autosomal dominant monogenic form of type 2 diabetes mellitus (DM) representing 5% of youth-onset DM in the Caucasian population. In young adults the disease can be present as either non insulin dependent or insulin-dependent DM. The diagnosis of this genetic disorder in children and adolescents is rare because of the mild glucose metabolism disorder at this time. We performed a metabolic, autoimmune and genetic study in 40 offspring of young parents affected by insulin-dependent DM (Group A) and in 35 offspring of young parents affected by early-onset non-insulin-dependent DM (Group B). Two children of Group A (5%) were found to be affected by fasting hyperglycemia and carry a GCK gene mutation that in one case was present also in the diabetic father. Eighteen offspring of Group B (51%) were positive for GCK or HNF-1alpha gene mutations present in the affected parents. All but two of these young patients had impaired fasting glucose (IFG) or impaired glucose tolerance (IGT). Eleven of them were younger than 16 years. We conclude that screening for DM in youth should be extended to MODY in young families with both non-insulin dependent and insulin-dependent DM. The sensitivity of the metabolic tests will precede the genetic diagnosis. PMID- 11393553 TI - Update on major trials for the prevention of type 1 diabetes mellitus: the American Diabetes Prevention Trial (DPT-1) and the European Nicotinamide Diabetes Intervention Trial (ENDIT). AB - The general population risk of developing type 1 diabetes mellitus (DM), 1/300, is magnified 15-20 fold in first-degree relatives of affected individuals. Because a combination of immunologic, metabolic, and genetic markers can be used to predict the disease, multicenter prevention trials in the US (DPT-1) and Europe (ENDIT) were initiated in relatives. In the DPT-1 over 80,000 relatives under 45 years of age will be screened for ICA and then 'staged' to assess risk. High-risk subjects (>50% over 5 yr) are randomized either to 4 days intravenous insulin infusion annually followed by b.i.d. low doses of subcutaneous ultralente insulin, or to close observation. To date (September 2000), 331/340 (97%) high risk subjects have been enrolled with the intention of detecting a 35% decrease in disease over 5 years (80% power). 280/490 (57%) of intermediate risk subjects (25-50% over 5 yr) have been randomized to oral insulin or placebo. A 50% treatment difference is sought. Anticipated enrolment for the high-risk arm will be completed by year 2001, and by 2003 for the oral arm. The ENDIT study will prospectively address whether nicotinamide will reduce the rate of progression to DM in relatives. 40,000 first-degree relatives (5-40 yr) have been screened with 552 subjects (ICA titers > or = 20 JDF U) randomized to nicotinamide or placebo. This study is designed with 90% power to detect a 35% reduction in disease (placebo group estimated at 40% risk over 5 years). Analysis of data is expected in 2003. PMID- 11393554 TI - Intervention to delay the onset of diabetes mellitus in very young siblings of children with type 1 diabetes mellitus. PMID- 11393555 TI - Changes in food habits in families with a newly diagnosed child with type 1 diabetes mellitus. AB - Dietary recommendations for children with type 1 diabetes mellitus (DM) are in line with the recommendations for the general population and applicable to the whole family. We review what is known about the food habits of non-diabetic family members and present original data on dietary changes in families with a child with DM. Some studies suggest that family members eat mostly the same food as the affected child. In a Finnish study of siblings of children with DM, favourable changes were observed after diagnosis in the type of milk and fat used. In a study of young children with DM, family members increased their consumption of skim milk, low-fat cheese and low-fat cold meat cuts. The consumption frequencies of fruit and vegetables increased. In conclusion, family members of a child with DM are willing to change their food habits towards the recommended diet. Dietary advice should be directed to the whole family from the very beginning. PMID- 11393556 TI - Psychological aspects of motherhood with type 1 diabetes mellitus--a personal experience. PMID- 11393557 TI - Fathers of children with diabetes mellitus and their role in coping strategies in the family. AB - Family structure and social disadvantage are thought to have adverse effects on the outcome of diabetes mellitus (DM). While there are data on family functioning and maternal coping in relation to some measures of metabolic control, little is known about the role of fathers in respect to coping and outcome of DM. We are presently conducting prospective studies to assess the role of fathers in families with children with DM. In addition, family functioning and psychosocial factors are being investigated in relation to putative effects on metabolic control. Structured questionnaire studies are being performed in 182 children and adolescents with type 1 DM. Similar questions are also put to the families. Mean age of the patients is 12.9 years, range 1.8-18 years, with an equal distribution between the sexes. Metabolic control as assessed by the mean of the last four HbA1c values (HPLC method; intra-assay coefficient of variation [CV] 2.1-3.3%, interassay CV 2.6-4.3%) is 7.4%; range 5.0-14.8%. The patients attend outpatient clinics at an average interval of six weeks. Structured educational in-patient programs are attended by less than one-third of the patients. There is a large prevalence of unemployment in the families. In the majority of cases mothers accompany their children to the clinic while fathers are absent. It is the mothers who adjust daily insulin doses, take care of food preparation and ensure the children's self-assessment. In conclusion, our preliminary data suggest that the role of fathers in diabetes management is rather passive and emotionally labile. Special educational programs targeted towards the fathers of children with DM and towards improving parental interactive strategies are urgently needed to better use parental resources of coping and support. PMID- 11393558 TI - Children with diabetes mellitus and family functioning: translating research into practice. AB - The specific challenges of living with diabetes mellitus (DM) which the child and parent must confront at each developmental stage are identified from infancy through adolescence. This discussion is divided into four developmental stages: 1) DM in infancy (0-2 years of age); 2) DM in toddlers and preschoolers (2-5 years); 3) DM in the school-age child (6-11 years); and 4) DM during early- to mid-adolescence (12-16 years). The central milestones of normal psychosocial development are reviewed and a summary is provided of recent research at each developmental stage with an emphasis on clinical implications for pediatric diabetes care teams. PMID- 11393559 TI - Screening programs to identify children at risk for diabetes mellitus: psychological impact on children and parents. AB - Screening programs to identify persons atrisk for diabetes mellitus (DM), before disease onset, are considered essential to understanding the natural history of the disease and for prevention program development. However, screening programs are complicated by imprecise markers of disease risk, the absence of a known effective prevention method, the use of children, and a wide variety of psychological, social, and educational challenges. Research relevant to four issues is presented: (1) parent and child anxiety in response to at-risk notification as well as how participants cope with this news; (2) accuracy of mothers' understanding of their babies' risk status; (3) predictors of participant recruitment and retention in longitudinal studies of this type; and (4) protocol compliance in prevention trials for type 1 DM. Integration of behavioral research into screening and prevention trials would help address the ethical concerns raised by such trials and improve their scientific quality. PMID- 11393560 TI - Early life stress and disease among offspring and siblings of individuals with type 1 diabetes mellitus. AB - Major life events, recent life stressors, and childhood diseases were examined among children and adolescents who were offspring, siblings, or other relatives of persons with type 1 diabetes mellitus (DM). All youth were recruited as part of a multi-site nationwide trial on the prevention of type 1 DM; parents of 347 children (4 to 18 yr) completed measures that asked about children's life events, recent stressors, and childhood illnesses. Analyses compared age groups (young child, preadolescent, adolescent) and relative type (offspring, sibling, other relative). Findings revealed offspring and siblings did not differ from "other relatives" in terms of life events, recent life stress, and disease/illness variables. However, siblings were reported to have fewer major life events and fewer life stressors in the past 12 months than offspring; siblings also had fewer infectious diseases during the first two years of life compared to offspring. Few age-related differences were found. Overall, results suggest that offspring and siblings of persons with type 1 DM are not at a disadvantage in terms of early life stress or disease in comparison to youth who have other family members with diabetes. However, siblings may have some advantages relative to children who are offspring. The mechanisms underlying these relationships require further elucidation and study. PMID- 11393561 TI - Psychological and ethical aspects of prevention trials. AB - Advances in immunological and genetic tests and in therapeutics may soon make it possible to predict, prevent or delay the development of type 1 diabetes mellitus (DM). Psychological support should be available at every stage of the screening and prevention process for high-risk individuals. Each step in this process results in a threat to the affected individual and his/her family, followed by anxiety, decision processes, uncertainty, conflicts and depression, and also attempts to cope with these. At the first step, information about the screening procedure is provided, and an attempt is made to motivate individuals and families to take part. At the second step, there is notification of risk and, for the small group of marker-positive individuals, an invitation to participate in a prevention trial. At the third step, the individuals included in a prevention protocol have to deal with anxiety over receiving placebo, maintain cooperation with therapy, and cope with potential failures of the prevention. There are only a few psychological data gained in DM screening or intervention studies, showing high clinical anxiety while screening, both before and after notification of risk. After a few months of anxiety, it usually drops to normal levels, but family functioning seems to be changed. Data are explained within the framework of an extended Health Belief Model. Data regarding the psychological impact on family members and identification of variables related to continued study participation are needed for studies seeking to recruit and retain subjects in longitudinal protocols for prediction and prevention of type 1 DM. PMID- 11393562 TI - Psychological impact of islet cell antibody screening. AB - The purpose of this study was to evaluate the psychological impact of autoantibody screening and its results on at-risk individuals and family members. Individuals who were antibody positive (AP) were identified through a large-scale screening program conducted at our institute. The sample consisted of nine families in whom 10 AP youngsters (7 M, 3 F) were identified, ranging in age from 6-18 years (mean 11.8, median 10 yr). Seventeen parents and eight diabetic youngsters (mean age 15.2, median 16 yr) participated in the study. Reaction to autoantibody positivity was assessed with the Impact of Event scale (IES). The IES was answered twice: within a week from the disclosure of the AP status, and 3 months later. Parents scored higher than their diabetic children and AP children on both measures of the IES, Intrusion and Avoidance. Three months later both scores were significantly reduced in both the parents and the AP children; however, parents still scored significantly higher on both scores than the AP children. The results suggest that learning one's AP status induces significant anxiety, especially in parents of AP youngsters. Although this initial anxiety dissipates over time it still remains quite high after 3 months. The results highlight the importance of psychosocial counseling for all members of diabetes mellitus screening and prevention trials. PMID- 11393563 TI - School-age children born to diabetic mothers and to mothers with gestational diabetes exhibit a high rate of inattention and fine and gross motor impairment. AB - OBJECTIVES: To study the neurobehavioral effects that pregestational and gestational diabetes might have on offspring at school age. STUDY DESIGN: We studied neurobehavioral function at school age of 57 children born to 48, well controlled, diabetic mothers and of 32 children born to 32 women with gestational diabetes. Their development was compared with 57 control children matched by age, birth order and parental socio-economic status, using a number of cognitive, behavioral, sensory and motor neurological tests. RESULTS: The IQ scores on the WISC-R and Bender tests of the children born to diabetic mothers were similar to those of control children, but the Bender scores of the children born to mothers with gestational diabetes were slightly lower. No differences were found between the study groups in various sensory-motor functions in comparison to controls. However, both study group children performed less well than controls in fine and gross motor functions as observed on the Bruininks-Oseretzki test of motor proficiency. The scores of children born to the mothers with pregestational diabetes were higher than controls on the Touwen & Prechtl neurological examination (worse function). The children from both research groups also performed worse on the Pollack taper test that is designed to detect inattention and hyperactivity. The children of the two study groups also had slightly higher scores on the Conners abbreviated parents-teachers questionnaire measuring hyperactivity and inattention, but the differences from controls were not statistically significant. We found a negative correlation between the performance of the children born to mothers with pregestational diabetes on various neurodevelopmental and behavioral tests and the severity of maternal hyperglycemia as assessed by blood glycosylated hemoglobin levels and acetonuria. CONCLUSIONS: Pregestational or gestational diabetes was found to adversely affect attention span and motor functions of offspring at school age, but not their cognitive ability. These effects were negatively correlated with the degree of maternal glycemic control, and were more pronounced in younger children. PMID- 11393564 TI - Health status of the children of diabetic mothers. PMID- 11393565 TI - Children of mothers with diabetes mellitus--a comment. PMID- 11393566 TI - Epilogue: what have we learned to improve the treatment of children with diabetes mellitus and their families? PMID- 11393567 TI - Update on endocrine disturbances in anorexia nervosa. AB - The marked endocrine changes that occur in anorexia nervosa have aroused a great deal of interest, and over the last decade much research has been conducted in this field. The endocrine disturbances are not specific to this disorder, as they also occur in starvation states secondary to other causes, and they return to normal upon weight restoration. However, emaciation may have profound effects on psychological processes, establishing an intricate circular interaction whereby somatic and psychological manifestations of starvation may continue to act. The purpose of this paper is to provide an overview of the large body of literature concerning endocrine aspects of anorexia nervosa with the main focus on the latest results, which provide leads for potential etiological theories. PMID- 11393568 TI - On longitudinal bone growth, short stature, and related matters: insights about cartilage physiology from the Utah paradigm. AB - Precursor cell division in growing cartilage determines human height, the lengths of the spine and limb bones, the alignment of joints, spines and limbs, and the ratio of spinal length to limb length. That division also helps to determine the sizes and shapes of joints, apophyses and epiphyses. Ideas about what controls those facts are changing. To former views, in which mainly genetic and humoral factors controlled them, the Utah paradigm of skeletal physiology adds long overlooked biomechanical including muscular factors. These three kinds of factors would collaborate in controlling the precursor cell division that determines the above skeletal features. Problems with that control clearly cause or help to cause many clinical disorders. Examples include short stature, gigantism, premature and delayed skeletal maturation, some changes in fracture patterns associated with puberty, joint malalignments, congenital hip dysplasia, scoliosis, limb torsions, the ball-and-socket ankle, and some skeletal abnormalities in Marfan's syndrome and the osteochondrodystrophies. The physiology such things depend on has matured sufficiently to justify a review for pediatricians, endocrinologists and other clinical specialists, and many basic scientists. PMID- 11393569 TI - A history of growth hormone injection devices. AB - In the early 1960s, growth hormone (GH) deficiency was treated by intramuscular injection of GH extracted from human pituitary glands. Since then, there have been many advances in treatment encompassing the route of administration, the injection product and the injection device. This review considers the advances in injection device that have already taken place and how they have benefited the patient, particularly in terms of reduced pain and improved convenience. In the future, needle-free injection techniques and depot formulations of GH are likely to offer alternatives to daily subcutaneous injections. PMID- 11393570 TI - Occurrence of thyroid papillary carcinoma in young patients. A Chernobyl connection? AB - The risk of thyroid papillary carcinoma is increased by external radiation particularly in children under 15 years of age as shown by a marked increase in those exposed to radiation after Chernobyl. We were recently confronted in Belgium over a short period with four patients (3 F, 1 M) with papillary thyroid carcinoma who were aged 10 years, 2 months, 2 years and 6 years when the Chernobyl accident occurred. We thus raise the question of a possible relationship. The patients were aged 17, 11, 10, 19 years at presentation. They all presented fortuitously over 3 years which was a very unusual increase in our extensive experience in thyroid surgery (62 cases of thyroid cancer among 1014 thyroidectomies in adults vs 4 cases in 18 children since the Chernobyl accident in 1986). Two out of the four patients had psammoma bodies (identifiable on CT scanning and ultrasound) and thyroglobulin autoantibodies (TgAb). The first patient had positive lymph nodes at the time of surgery. The incidence of thyroid cancers in Belarus and Ukraine rose just 4 years after the Chernobyl disaster; because radioactive clouds passed over Belgium, we wonder whether the occurrence of thyroid cancer in our patients could be related to this irradiation. The mechanism of increased incidence of radiation-induced thyroid cancer is thought to be due to rearrangement of the tyrosine kinase domains of the RET and TTK genes. The other important similarities in our patients are the presence of psammoma bodies that can be visualized on radiological examination and the presence of TgAb that are more frequent in differentiated thyroid cancers. Whether or not these cases reflect an increased incidence in the population as a whole, clinicians must remain vigilant for this rare but curable cancer in young patients, especially if suggestive radiological features or TgAb are present. PMID- 11393571 TI - Diabetic retinopathy in childhood: long-term follow-up by fluorescein angiography beginning in the first months of disease. AB - BACKGROUND: Little is known about minimal retinal lesions occurring in the first months of disease in children with type 1 diabetes mellitus (DM). OBJECTIVE: To detect any early retinal change and to evaluate its progression in children diagnosed with type 1 DM. PATIENTS: From 1979 to 1997 we examined by fluorescein angiography at diagnosis or within 15 months from the onset of DM 130 young patients with type 1 DM (mean age at diagnosis 10.08 +/- 2.62 yr). In 112 patients follow-up by fluorescein angiography was performed every 1.26 years with a mean of 5.41 fluorescein angiographies/patient. METHODS: The stage of retinopathy was graded to detect minimal lesions. We also considered sex, pubertal stage, HLA, family history of DM, disease duration and HbA1c levels. RESULTS: At first examination, 14 out of 127 (11%) readable angiographies showed minimal retinal changes. There was no statistically significant difference between the patients with or without lesions for all parameters considered. The 112 patients examined during follow-up were divided as follows: Group A: no retinopathy at first examination; Group A1: no retinopathy during follow-up; Group A2: retinal changes during follow-up; Group B: retinal changes at the first examination. Mean HbA1c value evaluated during the whole follow-up was lower in group A1 than in group A2. HbA1c levels at onset of the disease were significantly different in the three groups: in group A1 it was lower than in group A2 and in group B. CONCLUSIONS: The presence of early lesions in the first year of disease in 11% of patients is probably due to the method of examination, which may detect even minimal retinal changes. This may be correlated to the acute metabolic failure present at the onset of disease. The prolonged follow-up seems to demonstrate that the early changes are not necessarily a negative prognostic factor in the evolution of diabetic retinopathy. We confirm that duration of DM and metabolic control are the main factors influencing the course of retinopathy in these young patients. Early fluorescein angiography is not particularly useful in the management of children with DM. PMID- 11393572 TI - Diagnostic value of pituitary MRI in differentiation of children with normal growth hormone secretion, isolated growth hormone deficiency and multiple pituitary hormone deficiency. AB - Pituitary height, volume and morphology were investigated by MRI in patients aged 3.5-24.9 years with growth hormone deficiency (GHD) in relation to birth history and hormonal findings. Three groups with comparable age, sex and pubertal stage were studied: group I (n=42)--patients with isolated growth hormone deficiency (IGHD); group II (n=22)-- patients with multiple pituitary hormone deficiency (MPHD); and group III (n=30)--healthy controls. Pituitary height and volume differed significantly between the three groups, with the smallest in group II and largest in group III (p <0.001 for both). Both variables correlated significantly with peak GH value in the patient groups (p <0.001). The specificity of pituitary dysmorphology in the determination of GHD was 100% and its sensitivity in differentiation of IGHD and MPHD was 95%. Ectopic neurohypophysis was present in 75% of breech births and 27% of head-presenting patients (p <0.01). This study emphasizes the differential diagnostic value of pituitary MRI and its contribution to the understanding of the pathogenesis and prognosis in GHD. PMID- 11393573 TI - Decreased bone mineral density and bone formation markers shortly after diagnosis of clinical type 1 diabetes mellitus. AB - We recently demonstrated that children with type 1 diabetes mellitus (DM) have decreased lumbar spine bone mineral density (BMD) as early as four years after clinical diagnosis of the disease. In order to determine whether osteopenia is already present in patients very early on after diagnosis of clinical DM, we evaluated the bone mineral status of a group of newly diagnosed children (5.8 +/- 1.5 mo after diagnosis). We studied 23 prepubertal children (7 M, 16 F) with a mean chronological age of 9.5 +/- 2.2 yr and a mean glycosylated hemoglobin of 8.9 +/- 2.4%. Lumbar spine and femoral neck BMD were measured by dual X-ray absorptiometry, while bone turnover was assessed by the determination of the serum concentration of the carboxy-terminal propeptide of type I collagen (PICP) and the carboxy-terminal cross-linked telopeptide of type I collagen (N telopeptide). Results were compared to those of age, height, and pubertal status matched controls. Lumbar spine BMD Z-scores were decreased in patients compared to controls (Z-scores of -0.89 +/- 1.2, with 10 of 22 patients showing values >1 SD below the mean). When lumbar spine Z-scores were analyzed in those patients with <3 months or > or =3 months since diagnosis of DM a significant difference was noticed between groups (-0.648 +/- 1.12 vs -1.267 +/- 1.17; p <0.02). No significant differences were noted in femoral neck BMD and total BMD between groups. Serum PICP levels were decreased when compared to controls (233.6 +/- 39.3 vs 375.9 +/- 50.7 microg/l; p <0.002), while serum N-telopeptide concentrations, although increased, were not significantly different (9.3 +/- 1.3 vs 5.7 +/- 1.5 microg/l). In summary, early on after the diagnosis of type 1 DM, children present with decreased lumbar spine BMD and decreased bone formation markers. PMID- 11393574 TI - Efficacy of low dose schedule pamidronate infusion in children with osteogenesis imperfecta. AB - BACKGROUND: Osteogenesis imperfecta (OI) is a rare condition in which bones are abnormally brittle with frequent fractures. A variety of therapeutic agents has been used with low efficacy. In this study, we present three patients treated for 4 years with i.v. pamidronate. PATIENTS AND METHODS: Three prepubertal patients, aged 9 (M), 9 (F) and 11 (F) years old, with OI, were treated with 30-60 mg i.v. pamidronate every 6 months over four years. Determinations were made of plasma 1,25-dihydroxycholecalciferol, 25-hydroxycholecalciferol, insulin-like growth factor-I (IGF-I) and its transport protein (IGFBP3), osteocalcin, total alkaline phosphatase and its osseous fraction, and parathormone (PTH) at baseline and after every pamidronate infusion, Densitometry and X-ray of the vertebral column were performed at the same intervals. RESULTS: Significant reductions of number of bone fractures and pain were observed in all patients, despite lack of any modification in biochemical parameters. Lumbar X-ray and densitometry showed a striking improvement by the end of the treatment period. CONCLUSION: Pamidronate seems to be useful in the treatment of patients with osteogenesis imperfecta. PMID- 11393575 TI - The epidemiology of childhood-onset type 1 diabetes mellitus in Romania. ONROCAD Study Group. National Romanian Organisation for the Care of Diabetic Children and Adolescents. AB - The ONROCAD Study group was established to provide descriptive epidemiological information on type 1 diabetes mellitus (DM) in Romania. Data on all new patients with type 1 DM with onset before age 15 years during the four-year period 1992 1995 in Romania were submitted from all members of the ONROCAD Study Group, representing all doctors and clinics that manage children with DM in Romania. Age and sex-specific incidence rates were estimated, using available demographic data. A total of 706 new patients with type 1 DM and onset 0-14 years was registered. The overall completeness of ascertainment was estimated at 93.5%. For all Romania, the incidence of childhood-onset type 1 DM was estimated at 3.57/100,000/year. The incidence in the three regions was statistically significantly different (p = 0.002), with high rates in Transylvania and low rates in Muntenia. Girls had a significantly higher incidence than boys for each of the age groups 0-4, 5-9 and 10-14 years. Between these age groups, the incidence increased significantly with age. The incidence of childhood-onset type 1 DM is among the lowest recorded in Europe. Significant, but unexplained, geographical variation exists within Romania. Searches for factors to explain this heterogeneity may provide clues to the etiology of childhood-onset type 1 DM. PMID- 11393576 TI - Evaluation of the correlation between serum tumor necrosis factor-alpha and relative body mass index (RBMI) in childhood. AB - OBJECTIVE: Obesity and malnutrition are the main nutritional problems of childhood. Tumor necrosis factor-alpha (TNFalpha) is a well-known cytokine with effects on adipose tissue. In this study relative body mass index (RBMI) and serum TNFalpha levels were compared in obese and malnourished children, and hyperinsulinism was evaluated in the obese children. SUBJECTS AND METHODS: Thirty four children of whom 14 (9 M, 5 F) were obese, 10 (5 M, 5 F) were malnourished and 10 (4 M, 6 F) constituted the control group were evaluated. RBMI was used to define the groups according to the formula: patient's BMI/predicted BMI x 100. Mean RBMI values were 148.7 +/- 26, 77.6 +/- 4.63, and 100 +/- 8.2 in the obese, malnourished and control group, respectively. Mean ages of the obese, malnourished and control children were 9.75 +/- 2.59, 12.3 +/- 1.86 and 12.4 +/- 1.3 years, respectively. RESULTS: Higher serum TNFalpha levels were found in the obese children (20.94 +/- 0.38 pg/ml) in comparison with the control group (2.97 +/- 0.05 pg/ml) (p <0.001). Mean TNFalpha level in the malnourished group was 2.77 +/- 0.04 pg/ml, not statistically different from the control group (p >0.05). TNFalpha concentrations of the hyperinsulinemic and normoinsulinemic obese patients were not significantly different (p >0.05). CONCLUSION: We showed higher levels of serum TNFalpha in obese children. Increased TNFalpha levels do not always reflect hyperinsulinism. There was no difference in TNFalpha levels between the controls and the malnourished group. PMID- 11393577 TI - Helicobacter pylori infection in children with constitutional delay of growth and puberty. AB - Helicobacter pylori is a gastroduodenal pathogen strongly associated with chronic gastritis and duodenal ulceration. It is thought that H. pylori infection might be one of the causes of growth retardation in children. The aim of this study was to evaluate the seroprevalence of H. pylori in children with constitutional delay of growth and puberty (CDGP). H. pylori seropositivity was studied in 24 children with CDGP (22 M, 2 F) and 32 healthy age-matched children with normal pubertal development. Mean age of the children with CDGP was 14.53 +/- 1.12 yr and all of them had been diagnosed as CDGP after physical and laboratory assessment. H. pylori IgG positivity was detected in 16 of the 24 children with CDGP (66.6%) and 12 of 32 controls (37.5%) (p <0.05). This finding is consistent with the hypothesis that H. pylori infection could be one of the environmental factors causing CDGP. PMID- 11393578 TI - Clinical, endocrinological and radiography features in a child with McCune Albright syndrome and pituitary adenoma. AB - McCune-Albright syndrome is a rare syndrome presenting with polyostotic dysplasia, cafe-au-lait spots and multiple endocrinopathies that is very often combined with precocious puberty. We examined the clinical, endocrinological and radiological features in a boy with McCune-Albright syndrome and pituitary adenoma. X-rays, magnetic resonance (MRI) scan, whole body scintigraphy, single photon emission computer tomography (SPECT) and 3D-reconstruction from bone SPECT was performed to evaluate clinical improvement after treatment with sandostatin and pamidronic acid. After a six-month period of treatment with sandostatin and pamidronate, bone scintigraphy revealed significantly reduced activity. Treatment with bromocriptine and methimazole led to normalization of prolactin and thyroid hormone levels. Mobility of the patient improved. A significant improvement as a result of treatment with sandostatin and pamidronic acid was found in this patient with generalized fibrous dysplasia. So far, this condition has been treated with pamidronate only in adults, but severely affected children also benefit from this treatment regimen. PMID- 11393579 TI - Association of partial gonadal dysgenesis, nephropathy and WT1 gene mutation without Wilms' tumor: incomplete Denys-Drash syndrome. AB - The concurrence of ambiguous genitalia, nephropathy and predisposition to Wilms' tumor are characteristics of Denys-Drash syndrome. Some of the reported patients do not express the full spectrum of the syndrome, while the occurrence of nephropathy has become a generally accepted common feature of this syndrome. We report an infant with male pseudohermaphroditism due to partial gonadal dysgenesis and nephropathy without Wilms' tumor but with a Wilms' tumor suppressor gene (WT1) mutation. The high risk of Wilms' tumor mandates regular surveillance and the use of prophylactic bilateral nephrectomy as a treatment is not yet clear. PMID- 11393580 TI - Coexistence of common variable immunodeficiency and autoimmune polyglandular syndrome type 2. AB - A six year-old boy with common variable immunodeficiency developed insulin dependent diabetes mellitus, autoimmune thyroiditis, and total alopecia leading to the diagnosis of autoimmune polyglandular syndrome type 2. Previously unreported co-occurence of these two entities may be explained by strong autoimmunity and HLA association of both conditions. PMID- 11393581 TI - Cyproterone acetate: a genotoxic carcinogen? AB - Cyproterone acetate, a widely used synthetic progestagen with antiandrogenic activity, is known for years to produce liver tumours in rats, with a higher incidence in females. This effect was attributed to a rodent-specific tumour promoting mode of action based on the detection of a strong hepato-mitogenic activity of cyproterone acetate. However, more recent studies have demonstrated that cyproterone acetate is sex-specifically activated to (a) DNA-damaging intermediate(s) in the liver of female rats which result in the formation of DNA adducts, induction of DNA repair, and increased levels of micronuclei and gene mutations. Consistent with a sex-specific genotoxicity, cyproterone acetate showed a tumour-initiating potential in a liver foci assay with female rats but not with male rats. Most important, cyproterone acetate was found to induce formation of DNA adducts in primary cultures of human hepatocytes indicating that human liver cells have the capacity to activate cyproterone acetate to genotoxic intermediates. However, the overall assessment of the preclinical data presented in this review suggests that induction of liver tumours in female rats most probably depends on both, genotoxic and mitogenic effects which would suggest a non-linear mode of action with regard to tumour formation. With the exception of DNA adduct formation all other adverse effects induced by cyproterone acetate in rat liver, including gene mutations and liver tumours, can be detected at very high dose levels only. Hence, a cancer risk estimate based on a simple linear extrapolation from high dose to low exposure conditions of recommended clinical use would be questionable. Human data from pharmacoepidemiological studies that specifically addressed the question of possible liver cancer risk in patients treated with cyproterone acetate do in principle support this interpretation. In agreement with these considerations the regulatory authorities of the European Union came to the common conclusion that a possible cancer risk associated with the clinical use of cyproterone acetate, if any, appears to be low and the risk benefit ratios for the currently authorised indications remain favourable. PMID- 11393582 TI - Protective effect of berberine on cyclophosphamide-induced haemorrhagic cystitis in rats. AB - The urotoxicity of cyclophosphamide and the protective effect of the herb berberine were investigated in this study. Administration of 150 mg/kg cyclophosphamide intraperitoneally caused a serious haemorrhagic cystitis in rats after 12 hr, including bladder oedema, haemorrhage, and dramatic elevation of nitric oxide metabolites (nitrite+nitrate) in urine and in plasma. To explore whether cyclophosphamide-induced cystitis could be prevented by berberine, rats were pretreated with a single dose or two doses of berberine at 50, 100, or 200 mg/kg intraperitoneally then challenged with cyclophosphamide (150 mg/kg, intraperitoneally). The results indicated that pretreatment of rats with berberine could reduce cyclophosphamide-induced cystitis in a dose-dependent manner. Furthermore, we found that two doses of berberine showed greater protection against cyclophosphamide urotoxicity than when given a single dose. In addition, our data shows that a single dose of 200 mg/kg berberine, or two doses of 100, and 200 mg/kg berberine could completely block cyclophosphamide-induced bladder oedema and haemorrhage, as well as nitric oxide metabolites increase in rat urine and plasma. In conclusion, our findings suggest that berberine could be a potential effective drug in the treatment of cyclophosphamide-induced cystitis, and provides us with the bright hope in the prevention and treatment of cyclophosphamide urotoxicity. PMID- 11393584 TI - Diverging effects of 5-HT3 receptor antagonists ondansetron and granisetron on estramustine-inhibited cellular potassium transport. AB - We used 86Rb+ (K+ analogue) to study potassium influx during the interaction of highly specific 5-HT3-receptor antagonists, ondansetron and granisetron, with the effects of the anticancer drug, estramustine phosphate, on P31 mesothelioma cells. Estramustine phosphate (80 mg/l, 142 micromol/l) for 120 min. reduced 86Rb+ influx by 18.7%. The reduction was inhibited by ondansetron (0.1 micromol/l), but augmented by granisetron (0.1 micromol/l). Serotonin (1.0 micromol/l) antagonized ondansetron inhibition and restored granisetron-augmented reduction of estramustine phosphate-induced 86Rb+ influx to the level of the drug itself. Estramustine phosphate inhibited cellular Na+, K+, 2Cl- -cotransport activity whereas Na+, K+, ATPase activity was unaffected. Ondansetron blockade of estramustine phosphate-induced reduction of 86Rb+ influx was due to increased Na+, K+, ATPase and Na+, K+, 2Cl- -cotransport whereas augmentation of estramustine phosphate-induced reduction of 86Rb+ influx by granisetron, or combination of 5-HT3 receptor antagonists with serotonin was due mainly to inhibition of cellular Na+, K+, ATPase activity Thus, ondansetron possesses a distinct ability to reverse K+ influx of tumour cells exposed to estramustine phosphate whereas granisetron does not, due to different effect on cellular Na+, K+, ATPase and Na+, K+, 2Cl- -cotransport activity. Highly 5-HT3 receptor specific antiemetic agents may have different effects on ion transport of tumour cells during treatment with cytotoxic drugs. PMID- 11393583 TI - Central bromocriptine-induced tachycardia is reversed to bradycardia in conscious, deoxycorticosterone acetate-salt hypertensive rats. AB - A central dopaminergic origin has been demonstrated for the bromocriptine-induced tachycardia in conscious, normotensive rats. The present study investigated the effect of bromocriptine on heart rate and the principal site of action of this agonist in conscious, deoxycorticosterone acetate-salt hypertensive rats, in which altered central dopaminergic activity has been previously reported. Intravenous administration of bromocriptine (150 microg/kg) increased heart rate (49+/-5 beats/min.) in uninephrectomized control rats, while it induced a significant bradycardia (50+/-6 beats/min.) in deoxycorticosterone acetate-salt hypertensive rats. In the latter animals, intravenous (500 microg/kg) or intrathecal (40 microg/rat at T9-T10) pretreatment with domperidone, a selective dopamine D2 receptor antagonist that does not cross the blood-brain barrier, reduced partially, but significantly, the bradycardiac responses to bromocriptine (reduction of about 44% and 48% of the maximal effect, respectively). In contrast, the bromocriptine-induced bradycardia was fully abolished by intravenous pretreatment with metoclopramide (300 microg/kg), a dopamine D2 receptor antagonist that crosses the blood-brain barrier, or by combined pretreatment with intravenous and intrathecal domperidone. These results indicate that, in deoxycorticosterone acetate-salt hypertensive rats, bromocriptine decreases rather than increases heart rate, an effect that is mediated partly through a peripheral D2 dopaminergic mechanism and partly through stimulation of spinal dopamine D2 receptors. They further support the concept that, in normotensive, conscious rats, the central tachycardia of bromocriptine appears to predominate and to mask the bradycardia of this agonist at both peripheral and spinal dopamine D2 receptors. PMID- 11393585 TI - Role of daunorubicinol in daunorubicin-induced cardiotoxicity as evaluated with the model of isolated perfused rat heart. AB - Cardiotoxicity is the major side-effect and limits the clinical use of the anthracyclines, doxorubicin and daunorubicin. A special problem is raised by the metabolites of these drugs, the amount of which may vary according to drug combinations and formulations. Doxorubicinol, the 13-dihydroderivative of doxorubicin, has been shown to be more cardiotoxic than unchanged doxorubicin. Daunorubicinol has been assumed also to be more cardiotoxic than unchanged daunorubicin but its direct effect on cardiac function has never been evaluated in preclinical models. We have compared the cardiac effects (developed pressure, contractility and relaxation of the left ventricle) induced by daunorubicinol to those induced by daunorubicin, using the model of the isolated perfused rat heart. After treatment of rats with 6 doses of 3 mg/kg intraperitoneally, daunorubicin strongly decreased the cardiac functional parameters, while daunorubicinol did not induce cardiotoxicity. Both treatments induced a similar accumulation of daunorubicinol in the myocardium, while daunorubicin was only found in the hearts of rats treated with this drug. Direct perfusion of the hearts of untreated rats with both drugs at 10 microM induced a depression in heart function, but daunorubicin induced a progressive increase in diastolic pressure, reflecting the difficulties encountered by the heart to maintain its activity in the presence of this drug, while daunorubicinol did not. We conclude that daunorubicinol is not responsible for the important cardiac toxicity of daunorubicin. PMID- 11393586 TI - Microtiter-based assay for evaluating the biological activity of ribosome inactivating proteins. AB - A microtiter assay was developed to quickly measure the biological activity of ricin or other ribosome-inactivating proteins. Nuclease-treated rabbit reticulocyte lysate containing luciferase mRNA was used to measure toxin activity via inhibition of protein synthesis. The relative biological activity was determined by comparing luminescence levels in treated samples versus those of untreated controls. The amount of luciferase translated, as measured by luminescence, was inversely proportional to the toxin concentration. Linear dose response curves were generated for both class I and class II toxins. When compared to normal serum controls, specific antibody against ricin effectively inhibited ricin activity. The assay is suitable for efficient in vitro screening of therapeutics, as well as for identifying samples containing ribosome inactivating proteins. PMID- 11393587 TI - Butein, a plant polyphenol, induces apoptosis concomitant with increased caspase 3 activity, decreased Bcl-2 expression and increased Bax expression in HL-60 cells. AB - In the present study we have investigated whether butein could induce apoptosis in human leukaemic HL-60 cells. The treatment of HL-60 cells with butein induced apoptotic cell death as determined by morphological and biochemical changes. Apoptotic DNA fragments in the butein-treated HL-60 cells were increased gradually as determined by flow cytometric analysis. The caspase-3 activity was increased during butein-induced apoptosis. However, caspase-3 inhibitor abrogated the butein-induced DNA fragmentation. Furthermore, the treatment of HL-60 cells with butein decreased the expression of Bcl-2 protein, but increased the expression of Bax protein. These results suggest that butein-induced apoptosis is mediated through the activation of caspase-3 and it is associated with changed expression of Bcl-2 and Bax proteins. PMID- 11393589 TI - Nimesulide aggravates kainic acid-induced seizures in the rat. AB - Treatment of rats with kainic acid (10 mg/kg, intraperitoneally) triggers limbic seizures. Cyclooxygenase-2 mRNA is expressed in the hippocampus and cortex after 8 hr and marked cell loss occurs after 72 hr in the CA1-CA3 areas of the hippocampus. We examined the effect of the cyclooxygenase-2 inhibitor, nimesulide (N-(4-nitro-2-phenoxyphenyl)-methanesulfonamide), on kainate-induced seizures and delayed neurotoxicity. Nimesulide (10 mg/kg, intraperitoneally) was well tolerated given alone or 6-8 hr after kainate. However, pretreatment with nimesulide augmented seizures and increased the mortality rate from approximately 10% to 69%. We examined the effect of nimesulide on delayed cell loss after 72 hr in the surviving animals with histological staining. Cell loss did not seem to be reduced in animals treated with nimesulide 6-8 hr after kainate, but in the surviving animals pretreated with nimesulide less cell loss occurred. We conclude that nimesulide should be used with caution as an antiinflammatory drug in patients with convulsive disorders. PMID- 11393588 TI - Effects of genetic polymorphism of CYP1A2 inducibility on the steady-state plasma concentrations of trazodone and its active metabolite m-chlorophenylpiperazine in depressed Japanese patients. AB - The effects of a genetic polymorphism of inducibility of cytochrome P450 (CYP) 1A2 on the steady-state plasma concentrations of trazodone and its active metabolite, m-chlorophenylpiperazine, were studied in order to clarify if these steady-state plasma concentrations are dependent on the CYP1A2 polymorphism. Fifty-eight Japanese depressed patients received trazodone 150 mg/day at bedtime. The steady-state plasma concentrations of trazodone and m-chlorophenylpiperazine were measured in duplicate using high performance liquid chromatographic method, and were corrected to the mean body weight for analyses. A point mutation from guanine (wild type) to adenine (mutated type) at position -2964 in the 5' flanking region of CYP1A2 gene was identified by polymerase chain reaction fragment length polymorphism method. The mean steady-state plasma concentration of trazodone, but not m-chlorophenylpiperazine was significantly (P<0.05) lower in smokers than in non-smokers. Twenty-two smokers had 16 homozygotes of the wild type allele, 5 heterozygotes of the wild type and mutated alleles, and one homozygote of the mutated allele. There was no significant difference in the mean steady-state plasma concentration of trazodone or m-chlorophenylpiperazine between smokers with no mutation and those with mutation, although one homozygote of the mutated allele had the highest steady-state plasma concentration of trazodone in smokers. The present study thus suggests that CYP1A2 polymorphism does not necessarily have predictive value of the steady-state plasma concentration of trazodone or m-chlorophenylpiperazine in most of the smokers treated with trazodone. PMID- 11393590 TI - Beta-lactam antibiotic-mediated changes in platelet reactivity and vascular endothelial functions. AB - To evaluate vascular and platelet compatibility of intravenous administration of beta-lactam antibiotics, we assessed the effects of therapeutic concentrations of ceftriaxone, aztreonam, and ceftazidime on platelet reactivity to different agonists (sodium arachidonate, collagen and adenosine diphosphate) and on selected vascular endothelial functions (adenosine diphosphatase activity, prostacyclin production and t-PA release). Ceftriaxone and, to a lesser degree, aztreonam, enhanced platelet reactivity, evaluated as onset of platelet aggregating response, and increased thromboxane production to subthreshold concentrations of arachidonate. There was no modification in platelet reactivity after ceftazidime treatment. Ceftriaxone and ceftazidime, but not aztreonam, inhibited endothelial adenosine diphosphatase activity. Prostacyclin production and t-PA release were inhibited only by ceftriaxone at high concentrations. While it is difficult to establish which marker (platelet or endothelial functions) has more clinical reference in human vascular compatibility, it seems feasible to consider aztreonam the most compatible of the beta-lactams studied. PMID- 11393591 TI - Escitalopram (S-enantiomer of citalopram): clinical efficacy and onset of action predicted from a rat model. AB - Escitalopram is the active S-enantiomer of citalopram. In a chronic mild stress model of depression in rats, treatments with both escitalopram and citalopram were effective; however, a faster time to onset of efficacy compared to vehicle treatment was observed for escitalopram-treated (5 mg/kg/day) than for citalopram treated (10 mg/kg/day) rats at Week 1. To study the predictability of this observation in the clinic, we analysed 4-week data from an 8-week, double-blind, randomised, placebo-controlled, flexible-dose study that compared escitalopram and citalopram to placebo in primary care patients with major depressive disorder (baseline Montgomery and Asberg Depression Rating Scale (MADRS) scores > or =22 and < or =40). Since the flexible dosing started after Week 4, analysis of 4-week data ensured that the patients received fixed doses of 10 mg/day escitalopram (155 patients), 20 mg/day citalopram (160 patients), or placebo (154 patients). The efficacy analysis showed a significantly superior therapeutic effect for escitalopram versus placebo from Week 1 onwards (observed cases) with an adjusted mean change in MADRS at Week 4 (last observation carried forward) of 2.7 points (P=0.002). By comparison, 20 mg/day citalopram did not demonstrate a statistically significant effect compared to placebo. Escitalopram was well tolerated with an adverse event profile similar to that of citalopram. The preclinical observation that escitalopram possesses a faster time to onset of efficacy than citalopram was also seen in primary care patients with major depressive disorder. Thus, escitalopram is efficacious in depression and the effect occurs earlier than for citalopram. PMID- 11393592 TI - Psychotherapy outcome and quality improvement: introduction to the special section on patient-focused research. AB - This article introduces the special section of the Journal of Consulting and Clinical Psychology on patient-focused research for improving the outcome of psychological therapies. After a discussion of the context, promise, and problems associated with this research domain, an overview of the contributions is offered. PMID- 11393593 TI - Assessing treatment progress of individual patients using expected treatment response models. AB - The dosage model provides a normative estimate of the overall pattern of patient improvement in psychotherapy. The phase model further specifies patterns of change in the domains of subjective well-being, symptom remediation, and functioning. The expected treatment response (ETR) approach uses patient characteristics to predict an expected path of progress for each patient. With repeated measures of mental health status, the treatment progress of an individual patient can be assessed against the patient's ETR to support decisions that would enhance the quality of a clinical service while it is being delivered. PMID- 11393594 TI - Patient-focused research: using patient outcome data to enhance treatment effects. AB - A program of research aimed at improving the quality of psychological interventions is described. Data from over 10,000 patients were analyzed to understand the association between number of treatment sessions and clinically significant improvement. In addition to a potential dose-response relationship, typical recovery curves were generated for patients at varying levels of disturbance and were used to identify patients whose progress was less than expected ("signal" cases). The consequences of passing this information along to therapists were reported. Analyses of dose-response data showed that 50% of patients required 21 sessions of treatment before they met criteria for clinically significant improvement. Seventy-five percent of patients were predicted to improve only after receiving more than 40 treatment sessions in conjunction with other routine contacts, including medication in some cases. Identification of signal cases (potential treatment failures) shows promise as a decision support tool, although further research is needed to elucidate the nature of helpful feedback. Outgrowths of this research include its possible contribution to social policy decisions, reductions in the need for case management, use in supervision, and possible effects on theories of change. PMID- 11393595 TI - Computer-assisted feedback-driven quality management for psychotherapy: the Stuttgart-Heidelberg model. AB - The authors present the Stuttgart-Heidelberg Model for quality management of psychotherapy. The system is characterized as an active internal approach with external support from researchers. Problem-solving activities are initiated and maintained by computer-assisted feedback on possible treatment shortcomings. The system provides support for the 3 key tasks of quality management: (a) monitoring of relevant quality criteria in the clinical routine, (b) standardized assessment that allows comparison with established standards at various levels of service provision, and (c) transfer of information on achieved quality to those whom it concerns. The central function of the system is an alarm function, signaling a possible deficit of the provided psychotherapy on the basis of the standard evaluation of treatment outcome for individual patients. In this article, the main system components are described and data on its feasibility and validity are presented. PMID- 11393596 TI - Service profiling and outcomes benchmarking using the CORE-OM: toward practice based evidence in the psychological therapies. Clinical Outcomes in Routine Evaluation-Outcome Measures. AB - To complement the evidence-based practice paradigm, the authors argued for a core outcome measure to provide practice-based evidence for the psychological therapies. Utility requires instruments that are acceptable scientifically, as well as to service users, and a coordinated implementation of the measure at a national level. The development of the Clinical Outcomes in Routine Evaluation Outcome Measure (CORE-OM) is summarized. Data are presented across 39 secondary care services (n = 2,710) and within an intensively evaluated single service (n = 1,455). Results suggest that the CORE-OM is a valid and reliable measure for multiple settings and is acceptable to users and clinicians as well as policy makers. Baseline data levels of patient presenting problem severity, including risk, are reported in addition to outcome benchmarks that use the concept of reliable and clinically significant change. Basic quality improvement in outcomes for a single service is considered. PMID- 11393597 TI - Comparisons among quality assurance systems: from outcome assessment to clinical utility. AB - This special section describes contemporary systems for assessing the quality and effectiveness of service delivery. These systems have in common their commitment to the belief that by continuously monitoring treatment-related change, identifying problem cases, and providing feedback to clinicians or agencies regarding patient progress the benefits of treatment may be increased. Aside from their commonalities, much is to be learned from the varying ways with which these systems gather information and provide feedback to clinicians or health care managers. The methods vary both as a function of the sociopolitical climate of the country in which they were developed and of the personal preferences and assumptions of the developers. An articulation of these differences can be of interest to health care managers and to psychotherapy researchers. PMID- 11393598 TI - Cognitive versus behavior therapy in the group treatment of obsessive-compulsive disorder. AB - This study examined the effects of cognitive-behavior therapy (CBT) compared with traditional behavior therapy (exposure and response prevention [ERP]) in the group treatment of obsessive-compulsive disorder. Of the 76 participants who started treatment, 38 were wait-listed for 3 months before treatment to assess possible course effects. Both treatments were superior to the control condition in symptom reduction, with ERP being marginally more effective than CBT by end of treatment and again at 3-month follow-up. In terms of clinically significant improvement, treatment groups were equivalent on the conclusion of treatment, but 3 months later significantly more ERP participants met criteria for recovered status. Only 1 of 7 belief measures changed with treatment improvement, and the extent of this cognitive change was similar between CBT and ERP groups. Discussion includes consideration of optimal formats for the delivery of different types of treatment. PMID- 11393599 TI - Depressive symptoms and cigarette smoking among middle adolescents: prospective associations and intrapersonal and interpersonal influences. AB - Using data from a 4-wave longitudinal study with a school-based sample of 1,218 middle adolescents, the authors investigated the directionality (e.g., unidirectionality and bidirectionality) of the prospective relationship between depressive symptoms and cigarette use within the context of potential confounding variables and common and unique intrapersonal and interpersonal predictors. Findings indicated that serious and persistent depressive symptoms were prospective predictors of increased cigarette use across time, after controlling for baseline levels of smoking. Similarly, heavy and persistent smoking prospectively predicted increases in depressive symptoms. Intrapersonal and interpersonal predictors of cross-temporal changes in depressive symptoms and cigarette use were more unique than common. Latent growth curve modeling indicated a quadratic trend in adolescent cigarette smoking across time with an initial acceleration followed by a deceleration, though there was substantial intraindividual variation in individual trajectories. PMID- 11393600 TI - Relaxation and sleep compression for late-life insomnia: a placebo-controlled trial. AB - Older adults with insomnia were recruited from the community and randomized to treatments: relaxation, sleep compression, and placebo desensitization. Questionnaire data collected at baseline, posttreatment, and 1-year follow-up and polysomnography data collected at baseline and follow-up yielded the following conclusions: All treatments improved self-reported sleep, but objective sleep was unchanged. Clinical significance analyses yielded the strongest findings supporting the active treatments and suggested that sleep compression was most effective. Results partially supported the conclusion that individuals with high daytime impairment (i.e., fatigue) respond best to treatments that extend sleep, as in relaxation, and individuals with low daytime impairment respond best to treatments that consolidate sleep, as in sleep compression. Strong methodological features including a placebo condition and a treatment implementation scheme elevate the confidence due these findings. PMID- 11393601 TI - Specificity of relations between children's control-related beliefs and internalizing and externalizing psychopathology. AB - The authors examined the specificity of the relation between 3 types of control related beliefs and internalizing and externalizing psychopathology in a sample of 290 clinic-referred children aged 7 to 17 years. Self-reported beliefs about control (the capacity to cause an intended outcome), contingency (the degree to which a desired outcome can be controlled by a relevant behavior), and competence (an individual's ability to produce the relevant behavior) across 3 domains (academic, behavioral, and social) showed more specific relations with psychopathology than have been previously reported. Among children with externalizing psychopathology, internalizing psychopathology may be specifically associated with increased self-critical awareness about their conduct; externalizing psychopathology may attenuate the specific negative relation between internalizing psychopathology and control-related beliefs in the social domain. PMID- 11393602 TI - Childhood abuse and premilitary sexual assault in male Navy recruits. AB - Three samples of male U.S. Navy recruits (N = 7,850) were surveyed to determine whether a history of childhood physical abuse (CPA) or childhood sexual abuse (CSA) was predictive of premilitary rape of women and whether these relationships were mediated by alcohol problems and number of sex partners. In the 3 samples, 11.3%, 11.6%, and 9.9% of men reported committing premilitary rape. When demographic factors were controlled for, both CPA and CSA were independently and additively predictive of rape in each sample, with men who experienced both forms of abuse showing the highest risk of committing rape. Additional analyses revealed that alcohol problems and number of sex partners significantly mediated the relationship between childhood abuse (CPA and CSA) and rape perpetration. PMID- 11393603 TI - Predicting adherence to recommendations by parents of clinic-referred children. AB - The authors examined predictors of parents' adherence to recommendations made by psychologists after the evaluation of clinic-referred children. Parents or legal guardians of 93 children aged 4 to 12 years participated. The major findings were that (a) child behavior problem severity, parent recall of recommendations, parent satisfaction with the child's psychological evaluation, and locus of control were not significantly associated with adherence; (b) number of perceived barriers was the most salient predictor of adherence to recommendations, regardless of recommendation type; and (c) adherence rates to psychological services recommendations were significantly lower, compared with those for school based or professional nonpsychological recommendations. Implications for more research on predictors of adherence to recommendations and clinical strategies for overcoming barriers to adherence are discussed. PMID- 11393604 TI - Academic task persistence of normally achieving ADHD and control boys: performance, self-evaluations, and attributions. AB - The authors examined academic task persistence, pretask expectancies, self evaluations, and attributions of boys with attention-deficit/hyperactivity disorder (ADHD) as compared with control boys. Participants were 83 ADHD boys and 66 control boys, all normally achieving. Prior to the task, performance expectancies were assessed. After a success-failure manipulation with find-a-word puzzles, performance on subsequent trials, self-evaluations, and attributions were evaluated. Compared with controls, ADHD boys solved fewer test puzzles, quit working more often, and found fewer words on a generalization task. Consistent with these behavioral findings, research assistants rated ADHD boys as less effortful and less cooperative than control boys. Although ADHD boys did not differ significantly from controls in their posttask self-evaluations, they did differ significantly from controls in some aspects of their attributions. Attributional data indicated that ADHD boys endorsed luck as a reason for success more strongly and lack of effort as a reason for failure less strongly than controls. PMID- 11393605 TI - Depressed adolescents of depressed and nondepressed mothers: tests of an interpersonal impairment hypothesis. AB - Two groups of depressed youngsters were compared. From an interpersonal perspective, it was hypothesized that depressed adolescents of depressed mothers would have significantly more interpersonal dysfunction than depressed youngsters of nondepressed mothers. In a large community sample of youth and their families, 65 depressed offspring of women with histories of a major depressive episode or dysthymia were compared with 45 depressed offspring of never-depressed women. As predicted, after controlling for current symptoms and family social status variables, depressed offspring of depressed mothers displayed significantly more negative interpersonal behaviors and cognitions compared with depressed offspring of nondepressed mothers, but they did not differ on academic performance. Implications concerning mechanisms, course, and consequences of different forms of adolescent depression are presented. PMID- 11393606 TI - A controlled smoking cessation trial for substance-dependent inpatients. AB - Smoking treatment for newly recovering drug and alcohol-dependent smokers in a residential rehabilitation program was examined. The randomly assigned conditions (n = 50 each) were multicomponent smoking treatment (MST), MST plus generalization training of smoking cessation to drug and alcohol cessation (MST+G), or usual care (UC). Fifty participants who declined smoking treatment (treatment refusers) also were studied. Both treatment conditions achieved continuous smoking abstinence rates (MST: 12%, MST+G: 10%, at 12-month follow-up) that were significantly higher than in the UC condition (0%). The MST condition had a continuous drug and alcohol abstinence rate that was significantly higher than that of the MST+G condition (40% vs. 20% at 12-month follow-up) although neither differed significantly from that of the UC condition (33%). These results support the feasibility of smoking treatment for this population and provide information regarding appropriate treatment components. PMID- 11393607 TI - Eye movement desensitization and reprocessing (EMDR): a meta-analysis. AB - Eye movement desensitization and reprocessing (EMDR), a controversial treatment suggested for posttraumatic stress disorder (PTSD) and other conditions, was evaluated in a meta-analysis of 34 studies that examined EMDR with a variety of populations and measures. Process and outcome measures were examined separately. and EMDR showed an effect on both when compared with no treatment and with therapies not using exposure to anxiety-provoking stimuli and in pre post EMDR comparisons. However, no significant effect was found when EMDR was compared with other exposure techniques. No incremental effect of eye movements was noted when EMDR was compared with the same procedure without them. R. J. DeRubeis and P. Crits-Christoph (1998) noted that EMDR is a potentially effective treatment for noncombat PTSD. but studies that examined such patient groups did not give clear support to this. In sum, EMDR appears to be no more effective than other exposure techniques, and evidence suggests that the eye movements integral to the treatment, and to its name, are unnecessary. PMID- 11393609 TI - Child custody mediation and litigation: custody, contact, and coparenting 12 years after initial dispute resolution. AB - Long-term follow-up data were obtained on families who had been randomly assigned to mediate or litigate their child custody disputes. In comparison with families who litigated custody, nonresidential parents who mediated were more involved in multiple areas of their children's lives, maintained more contact with their children, and had a greater influence in coparenting 12 years after the resolution of their custody disputes. The increased involvement of nonresidential parents who mediated did not lead to an associated increase in coparenting conflict. Parents who mediated also made more changes in their children's living arrangements over the years. For the most part, the changes apparently reflect increased cooperation and flexibility. Satisfaction declined for parents (especially fathers) in both groups over time, but fathers remained much more satisfied if they mediated rather than litigated custody. Few differences in satisfaction were found between mothers in the 2 groups. The 12-year follow-up data indicate that, even in contested cases. mediation encourages both parents to remain involved in their children's lives after divorce without increasing coparenting conflict. PMID- 11393608 TI - A comparison of different methods for assessing the features of eating disorders in patients with binge eating disorder. AB - The authors compared 3 methods for assessing the features of eating disorders in patients with binge eating disorder (BED). Participants were administered the Eating Disorder Examination (EDE) interview and completed the EDE Questionnaire (EDE-Q) at baseline. Participants prospectively self-monitored their eating behaviors daily for 4 weeks and then completed another EDE-Q. The EDE and the EDE Q were significantly correlated on frequencies of objective bulimic episodes (binge eating) and on the Dietary Restraint, Eating Concern, Weight Concern, and Shape Concern subscales. Mean differences in the EDE and EDE-Q frequencies of objective bulimic episodes were not significant, but scores on the 4 subscales differed significantly, with the EDE-Q yielding higher scores. At 4 weeks, the EDE-Q retrospective 28-day assessment was significantly correlated with the prospective daily self-monitoring records for frequency of objective bulimic episodes, and the mean difference between methods was not significant. The EDE-Q and self-monitoring findings for subjective bulimic episodes and objective overeating differed significantly. Thus, in patients with BED, the 3 assessment methods showed some acceptable convergence, most notably for objective bulimic episodes. PMID- 11393610 TI - Structure of anxiety symptoms in urban children: competing factor models of the Revised Children's Manifest Anxiety Scale. AB - The Revised Children's Manifest Anxiety Scale (RCMAS; C. R. Reynolds & B. O. Richmond, 1985) is among the most widely used self-report measures of children's anxiety. The authors compared its current empirically derived factor structure with theory-driven models derived from 8 experts on child anxiety using concept mapping. Confirmatory factor analyses compared models using data from 898 seventh graders in an urban public school system serving a high percentage of African Americans. The most parsimonious best-fitting model was an expert-derived model with factors reflecting anxious arousal, social evaluation-oversensitivity, worry, and a higher order factor. This model was theoretically meaningful, excluded items less relevant to anxiety, and was invariant across gender. Future research with the RCMAS should consider use of these dimensions. The combination of qualitative and quantitative methodology used in this study appeared to have considerable utility for refining measures. PMID- 11393611 TI - Screening and treatment of distress. PMID- 11393612 TI - Influence of Stiles-Crawford effect apodization on spatial visual performance with decentered pupils. AB - Using theoretical estimates of the optical-transfer function and line-spread function as image-quality criteria, we predicted the influence of the Stiles Crawford effect (SCE) on both optical performance of the eye and subjective measurements of transverse aberrations when pupils are decentered. The SCE was modeled as a pupil apodization. The SCE appears to improve image quality by providing compensation for aberrations induced by pupil decentration, but this improvement is usually small. When a criterion of the placement of the image is used as the centroid of the line-spread function, an average SCE reduces the influence of pupil decentration on subjective transverse chromatic aberrations (TCA's) for 5-mm-diameter pupils by 30%. This reduction is much less than that obtained by previous experimental studies of TCA, and possible reasons for this discrepancy are discussed. Decentering the SCE produces an appreciable shift in subjective TCA for 5-mm-diameter pupils of 1.4 arc min per 1-mm decentration (at wavelengths 433 and 622 nm). PMID- 11393613 TI - Macular pigment density measured by autofluorescence spectrometry: comparison with reflectometry and heterochromatic flicker photometry. AB - We present a technique for estimating the density of the human macular pigment noninvasively that takes advantage of the autofluorescence of lipofuscin, which is normally present in the human retinal pigment epithelium. By measuring the intensity of fluorescence at 710 nm, where macular pigment has essentially zero absorption, and stimulating the fluorescence with two wavelengths, one well absorbed by macular pigment and the other minimally absorbed by macular pigment, we can make accurate single-pass measurements of the macular pigment density. We used the technique to measure macular pigment density in a group of 159 subjects with normal retinal status ranging in age between 15 and 80 years. Average macular pigment density was 0.48 +/- 0.16 density unit (D.U.) for a 2 degrees diameter test field. We show that these estimates are highly correlated with reflectometric (mean: 0.23 +/- 0.07 D.U.) and psychophysical (mean: 0.37 +/- 0.26 D.U.; obtained by heterochromatic flicker photometry) estimates of macular pigment in the same subjects, despite the fact that systematic differences in the estimated density exist between techniques. Repeat measurements over both short- and long-time intervals indicate that the autofluorescence technique is reproducible: The mean absolute difference between estimates was less than 0.05 D.U., superior to the reproducibility obtained by reflectometry and flicker photometry. To understand the systematic differences between density estimates obtained from the different methods, we analyzed the underlying assumptions of each technique. Specifically, we looked at the effect of self-screening by visual pigment, the effect of changes in optical property of the deeper retinal layers, including the role of retinal pigmented epithelium melanin, and the role of secondary fluorophores and reflectors in the anterior layers of the retina. PMID- 11393614 TI - Blur tolerance for luminance and chromatic stimuli. AB - We investigated the blur tolerance of human observers for stimuli modulated along the isoluminant red-green, the isoluminant yellow-blue, and the luminance (black white) direction in color space. We report the following results: (i) Blur difference thresholds for red-green and luminance stimuli (of equal cone contrast) are very similar and as low as 0.5 min of visual angle; for yellow-blue the lowest blur thresholds are much higher (1.5 min of visual angle). (ii) The smallest blur thresholds are found for slightly blurred square waves (reference blur of 1 arc min) and not for sharp edges. (iii) Blur thresholds for red-green and black-white follow a Weber law for reference (pedestal) blurs greater than the optimum blur. (iv) Using the model proposed by Watt and Morgan [Vision Res. 24, 1387 (1984)] we estimated the internal blur of the visual system for the black-white and the red-green color directions and arrived at the following estimates: 1.2 arc min for black-white stimuli at 10% contrast and 0.9 arc min for red-green stimuli at 10% cone contrast. Blur tolerance for yellow-blue is independent of external blur and cannot be predicted by the model. (v) The contrast dependence of blur sensitivity is similar for red-green and luminance modulations (slopes of -0.15 and -0.16 in log-log coordinates, respectively) and slightly stronger for yellow-blue (slope = -0.75). Blur discrimination thresholds are not predicted by the contrast sensitivity function of the visual system. Our findings are useful for predicting blur tolerance for complex images and provide a spatial frequency cutoff point when Gaussian low-pass filters are used for noise removal in colored images. They are also useful as a baseline for the study of visual disorders such as amblyopia. PMID- 11393615 TI - Importance of color in the segmentation of variegated surfaces. AB - We examined how variations in color and brightness are used by the visual system in distinguishing textured surfaces that differed in their first- or second-order statistics. Observers viewed a 32 x 32 array containing two types of square elements differing in chromaticity or luminance or both. The spatial distributions of the two kinds of elements were varied within the array until observers could distinguish two juxtaposed regions. At low but not at high contrast, observers are better able to distinguish regions when the elements differ only in chromaticity than when they differ only in luminance. The advantage of color at low contrasts results from the greater visibility of the arrays defined by color variation. An observer's capacity to distinguish textures defined by variations in first-order chromatic statistics is little affected by the addition of achromatic noise but is more affected by the addition of chromatic noise. The relative robustness of chromatic cues in the face of achromatic noise leaves the visual system well equipped to exploit color variations in segmenting complex scenes, even in the presence of variations in brightness. This capacity seems to depend on mechanisms that sum over large regions: When surfaces differ in their second-order statistics and cannot be distinguished by mechanisms that sum over large regions, the advantage of color is much diminished. PMID- 11393616 TI - Fractional Talbot field and of finite gratings: compact analytical formulation. AB - We present a compact analytical formulation for the fractional Talbot effect at the paraxial domain of a finite grating. Our results show that laterally shifted distorted images of the grating basic cell form the Fresnel field at a fractional Talbot plane of the grating. Our formulas give the positions of those images and show that they are given by the convolution of the nondistorted cells (modulated by a quadratic phase factor) with the Fourier transform of the finite-grating pupil. PMID- 11393617 TI - Effects of abrupt surface-profile transitions in nonparaxial diffractive optics. AB - It is shown that the failure of the thin-element approximation of diffractive optics may, in the first approximation, be attributed to local diffraction effects caused by the abrupt vertical transitions in binary surface-relief profiles. We determine the field disturbance caused by a single-step transition (of given height) by rigorous diffraction theory. Associating such a disturbance with each individual transition point in the profile, we obtain a computationally efficient refinement of the thin-element approximation for the analysis and design of diffractive elements in the nonparaxial domain. The results agree well with those obtained by global application of rigorous diffraction theory, provided that the smallest features in the binary profile are larger than approximately one optical wavelength. PMID- 11393618 TI - Nonpolarizing guided-mode resonant grating filter for oblique incidence. AB - A new type of guided-mode resonant grating filter is described. The filter is independent of polarization state for oblique incidence. The filter has a crossed grating structure, and the plane of incidence on the filter contains the symmetric axis of the grating structure. Theoretical considerations and numerical calculations using two-dimensional rigorous coupled-wave analysis show that a rhombic lattice structure is suitable to such filters. In this configuration an incident light wave is diffracted into the waveguide and is divided into two propagation modes whose directions are symmetric with respect to the plane of incidence. In particular, when the propagation directions of the two modes are perpendicular to each other, the fill factor of grating structure can be approximately 50%. The filter was designed for an incident angle of 45 degrees. Tolerances of setting errors and fabrication errors for this filter were estimated by numerical calculations. PMID- 11393619 TI - Statistical interferometry based on a fully developed speckle field: an experimental demonstration with noise analysis. AB - A novel interferometric method named statistical interferometry is proposed and studied. In the method, in contrast to the conventional deterministic interferometry, the complete randomness of the two interfering light fields, i.e., the random interference of the fully developed speckle fields, plays an essential role and is used as a standard of phase in a statistical sense. Preliminary experiments were conducted to verify the validity of the method, followed by a computer simulation. As an experimental result, the accuracy of the measurements of an out-of-plane displacement was confirmed up to lambda/800 by comparison with the heterodyne interferometer. The method has the advantage of simplicity of the optical system required, while at the same time providing high accuracy. PMID- 11393620 TI - Mie-scattering formalism for spherical particles embedded in an absorbing medium. AB - Most of the Mie-scattering calculations have been done for a particle embedded in a nonabsorbing host medium. Generalization to an absorbing host medium can be achieved (a) by modifying the calculation of the spherical Bessel functions to account for a complex argument and (b) by accounting properly for the net rate of incident, scattered, and absorbed energy. We present an extended formalism of Mie scattering for the case of an absorbing host medium. Numerical calculations show that for a large spherical particle embedded in an absorbing host medium the extinction efficiency approaches 1 compared with 2 for a nonabsorbing host medium. We conjecture that this difference is due to the suppression of diffraction when the radius of the sphere is large. PMID- 11393621 TI - Polarized light scattering by microroughness and small defects in dielectric layers. AB - The polarization of light scattered by the surface of a material contains information that can be used to identify the sources of that scatter. Theories for light scattering from interfacial roughness of a dielectric layer and from defects in that dielectric layer are reviewed. Methods for calculating the Mueller matrix or the Stokes vector for scatter from multiple sources and for decomposing a Stokes vector into contributions from two nondepolarizing scattering sources are derived. The theories are evaluated for a specific sample and geometry. Results show that some incident polarizations are more effective than others at discriminating among scattering sources, with s-polarized light being least effective. The polarization of light scattered from interfacial roughness depends upon the relative roughness of the two interfaces and the degree of correlation between the two interfaces. The scattering from defects in the film depends on the depth of the defect and differs from that from any one of the cases of interfacial roughness. The scattering from defects randomly distributed in the film and for small dielectric permittivity variations in the film is also calculated. Experimental results are presented for a 52-nm SiO2 film thermally grown on microrough silicon. PMID- 11393622 TI - Adaptive optics with advanced phase-contrast techniques. I. High-resolution wave front sensing. AB - High-resolution phase-contrast wave-front sensors based on phase spatial light modulators and micromirror/ liquid-crystal arrays are introduced. Wave-front sensor performance is analyzed for atmospheric-turbulence-induced phase distortions described by the Kolmogorov and the Andrews models. A high-resolution phase-contrast wave-front sensor (nonlinear Zernike filter) based on an optically controlled liquid-crystal phase spatial light modulator is experimentally demonstrated. The results demonstrate high-resolution visualization of dynamically changing phase distortions within the sensor time response of approximately 10 ms. PMID- 11393624 TI - Anisoplanatic imaging through turbulent media: image recovery by local information fusion from a set of short-exposure images. AB - A wide-field-of-view white-light imaging experiment with artificially generated turbulence layers located between the extended object and the imaging system is described. Relocation of the turbulence sources along the imaging path allowed the creation of controllable anisoplanatic effects. We demonstrate that the recently proposed synthetic imaging technique [J. Opt. Soc. Am. A 16, 1623 (1999)] may result in substantial improvement in image quality for highly anisoplanatic conditions. It is shown that for multisource objects located at different distances the processing of turbulence-degraded short-exposure images may lead to a synthetic image that has an image quality superior to that of the undistorted image obtained in the absence of turbulence (turbulence-induced image quality enhancement). PMID- 11393623 TI - Adaptive optics with advanced phase-contrast techniques. II. High-resolution wave front control. AB - A wave-front control paradigm based on gradient-flow optimization is analyzed. In adaptive systems with gradient-flow dynamics, the output of the wave-front sensor is used to directly control high-resolution wavefront correctors without the need for wave-front phase reconstruction (direct-control systems). Here, adaptive direct-control systems with advanced phase-contrast wave-front sensors are analyzed theoretically, through numerical simulations, and experimentally. Adaptive system performance is studied for atmospheric-turbulence-induced phase distortions in the presence of input field intensity scintillations. The results demonstrate the effectiveness of this approach for high-resolution adaptive optics. PMID- 11393625 TI - Color and spectral analysis of daylight in southern Europe. AB - We have analyzed the colorimetric and spectral characteristics of 2600 daylight spectra (global spectral irradiances on a horizontal surface) measured for all sky states during a 2-year period at Granada, Spain. We describe in detail the chromaticity coordinates, correlated color temperatures (CCT), luminous efficacies, and relative UV and IR contents of Granada daylight. The chromaticity coordinates of Granada daylight lie far above the CIE locus at high CCTs (>9,000 K), and a CCT of 5,700 K best typifies this daylight. Our principal-components analysis shows that Granada daylight spectra can be adequately represented by using six-dimensional linear models in the visible, whereas seven-dimensional models are required if we include the UV or near-IR. Yet on average only three dimensional models are needed to reconstruct spectra that are colorimetrically indistinguishable from the original spectra. PMID- 11393626 TI - Inverse problem in optical diffusion tomography. I. Fourier-Laplace inversion formulas. AB - We consider the inverse problem of reconstructing the absorption and diffusion coefficients of an inhomogeneous highly scattering medium probed by diffuse light. Inversion formulas based on the Fourier-Laplace transform are used to establish the existence and uniqueness of solutions to this problem in planar, cylindrical, and spherical geometries. PMID- 11393627 TI - Propagation of polychromatic Gaussian beams through thin lenses. AB - The transformation by a lens of a polychromatic laser beam composed of on-axis superposed monochromatic TEM00 Gaussian modes in the paraxial approximation is studied. The chromatic aberrations are described by allowing the waist position on the z axis and the Rayleigh range to depend on wavelength. The beam radius, the far-field divergence, the Rayleigh range, the beam product, the beam propagation factor, and the kurtosis parameter are calculated. The relationship between the fourth-order and the second-order moments of Hermite-Gaussian and Laguerre-Gaussian modes is obtained and is used for calculating kurtosis parameter. The results are generalized to polychromatic modes of higher orders. It is shown that the on-axis superposition of monochromatic TEM00 modes with no chromatic aberration is leptokurtic. PMID- 11393628 TI - Using rays better. III. Error estimates and illustrative applications in smooth media. AB - A new method for computing ray-based approximations to optical wave fields is demonstrated through simple examples involving wave propagation in free space and in a gradient-index waveguide. The analytic solutions that exist for these cases make it easy to compare the new estimates with exact results. A particularly simple RMS error estimate is developed here, and corrections to the basic field estimate are also discussed and tested. A key step for any ray-based method is the choice of a family of rays to be associated with the initial wave field. We show that, for maximal accuracy, not only must the initial field be considered in choosing the rays, but so too must the medium that is to carry the wave. PMID- 11393629 TI - Variational approach to moire pattern synthesis. AB - Moire phenomena occur when two or more images are nonlinearly combined to create a new superposition image. Moire patterns are patterns that do not exist in any of the original images but appear in the superposition image, for example as the result of a multiplicative superposition rule. The topic of moire pattern synthesis deals with creating images that when superimposed will reveal certain desired moire patterns. Conditions that ensure that a desired moire pattern will be present in the superposition of two images are known; however, they do not specify these images uniquely. The freedom in choosing the superimposed images can be exploited to produce various degrees of visibility and ensure desired properties. Performance criteria for the images that measure when one superposition is better than another are introduced. These criteria are based on the visibility of the moire patterns to the human visual system and on the digitization that takes place when the images are presented on discrete displays. We propose to resolve the freedom in moire synthesis by choosing the images that optimize the chosen criteria. PMID- 11393630 TI - Analysis of integrated optical waveguides: variational method and effective-index method with built-in perturbation correction. AB - We show analytically that for the modal analysis of rectangular waveguides, the recently developed effective-index method with built-in perturbation correction is identical to the first-iteration cycle of the variational method that we had developed earlier [Opt. Quantum Electron. 12, 517 (1989)]. We also show that in a number of cases the accuracy improves considerably through iteration beyond the first cycle. PMID- 11393631 TI - Optical properties of an isotrophic optically active medium at oblique incidence. AB - The optical properties of an isotropic optically active medium at oblique incidence have been investigated. It was found that the amount of transmitted light converted from p polarization to s polarization and vice versa, through an isotropic optically active medium, is independent of the state of incident polarization. Though the optical rotation through the optically active medium is same for p and s polarization at normal incidence, it becomes different at oblique incidences. PMID- 11393632 TI - Effect of beam divergence from the optic axis in an electro-optic experiment to measure an induced Jones birefringence. AB - Certain optical properties can be described in terms of two linear birefringences existing in separate Jones platelets of a medium. One of these, known as Jones birefringence, although occurring naturally in some crystals is too small to be measurable. However, the two birefringences can be induced by an electric field in 4 and 6 crystals for propagation along the optic axis. For an even slightly divergent light beam, natural birefringence may affect accuracy of measurement. Calculations show that in an experiment with a static field the error depends critically on beam divergence, whereas with a modulated field this is not so. PMID- 11393633 TI - Gaussian Schell-model beams propagating through polarization gratings. AB - The effects of polarization gratings on partially coherent beams are investigated by studying a Gaussian Schell-model beam impinging on a linear polarizer whose transmission axis varies periodically along one transverse direction. Analytical expressions for the beam polarization-coherence matrix after the grating are obtained. In particular, the evolution of the degree of polarization upon propagation is analyzed. Different behaviors of the output beam, depending on the beam parameters and on the period of the grating, are exhibited. In particular, it is shown that, by suitably choosing the latter quantities, it is possible to obtain not only any desirable value of the degree of polarization of the output beam but also particular distributions of such parameters across the transverse sections of the beam. PMID- 11393634 TI - Reflectance enhancement with sub-quarterwave multilayers of highly absorbing materials. AB - A new theory of multilayers with enhanced normal reflectance has been developed based on the superposition of a few layers of various different radiation absorbing materials. Every layer in the multilayer had a subquarterwave optical thickness. The theory was developed for materials with small refractive-index differences, although it is also valid in some cases for materials with large refractive-index differences. Reflectance enhancements were obtained in a very broad band and over a wide range of incidence angles. The theory is particularly suited to designing multilayers with enhanced reflectance in the extreme ultraviolet for wavelengths above 50 nm. In this spectral region the reflectance of single layers of all materials is relatively low, and standard multilayers are not possible because of the high absorption of materials. PMID- 11393635 TI - Dyspnea and the affective response during exercise training in obstructive pulmonary disease. AB - BACKGROUND: Dyspnea (SOB), dyspnea-related anxiety (DA), and exercise performance have been shown to improve after exercise training in patients with Chronic Obstructive Pulmonary Disease (COPD). However, there are no published descriptions of the changes in dyspnea intensity or dyspnea-related anxiety during or across the exercise training sessions. OBJECTIVES: To describe and compare the differences in the patterns of change in SOB, DA, and exercise performance during 12 exercise training sessions with and without nurse coaching. METHODS: Forty-five dyspnea-limited patients with COPD were randomly assigned to nurse-monitored (ME) or nurse-coached exercise (CE). SOB and DA were rated on a 200 mm VAS every 2 minutes during each of 12 treadmill training sessions. RESULTS: Warm-up, peak, cool-down, mean SOB, and peak SOB/stage remained constant over the exercise sessions, with increasing exercise performance for both groups over the 12 sessions (p < .001). There was a significant difference in the pattern of mean SOB over time between the ME and CE group (p < . 05). Mean, peak DA, and peak DA/stage showed a rapid decrease within the first 4 sessions (p < . 05) with no significant differences between the groups. Warm-up and cool-down DA remained constant. There were large intra- and inter-subject variations in the rating of dyspnea and dyspnea-related anxiety within and across sessions. CONCLUSIONS: As theoretically proposed, both groups significantly decreased their DA over the training sessions. This decrease was early in the sessions and was not accompanied by a decrease in the SOB. In contrast, subjects maintained a nearly constant mean and peak SOB with increasing exercise performance, suggesting that people may have a dyspnea threshold above which they are unable to tolerate greater dyspnea. Description of the changes in dyspnea and the affective response during training need to be expanded, while studying the type and timing of strategies to enhance the improvement in dyspnea and dyspnea related anxiety. PMID- 11393636 TI - Emotional distress and activities of daily living functioning in persons with multiple sclerosis. AB - BACKGROUND: Emotional distress is higher in persons with multiple sclerosis (MS) than in other chronic illnesses. Not known is whether personal attributes of the person with MS and/or the presence of social support will function as mediating and/or moderating variables between emotional distress and adaptation to the illness. OBJECTIVES: Determine if personal attributes and social support function as mediating and/or moderating variables between emotional distress and ADL functioning in persons with MS. METHODS: Secondary analyses of data obtained from 686 persons with MS through self-report measures of emotional distress, personal attributes, social support, and ADL functioning was conducted. Separate mediation and moderation models were tested using stepwise and hierarchical multiple regression. Demographic variables of education, age, and length of MS illness, were controlled in all analyses. RESULTS: Personal attributes and social support functioned as mediator variables between emotional distress and ADL functioning. Additionally, personal attributes and not social support functioned as a moderator. Significant main effects were shown for social support and emotional distress in the moderator model. CONCLUSION: Personal attributes and social support mediated the effects of emotional distress by decreasing its impact on ADL functioning. Personal attributes, as a moderator variable, demonstrated that higher levels were associated with low levels of emotional stress and moderate or lower levels of personal attributes were associated with increased emotional distress suggesting that personal attributes may intervene between emotional distress and ADL functioning by attenuating or preventing a stress appraisal response. PMID- 11393637 TI - Further validation of the Postpartum Depression Screening Scale. AB - BACKGROUND: Up to 50% of all cases of postpartum depression go undetected. The Edinburgh Postnatal Depression Scale (EPDS) has been the only instrument available that was specifically designed to screen for this mood disorder. None of the items on the EPDS, however, are written in the context of new motherhood. OBJECTIVE: The purpose of this study was to further assess the construct validity of the newly designed Postpartum Depression Screening Scale (PDSS) along with its sensitivity, specificity, and predictive values. METHOD: A total sample of 150 mothers within 12 weeks postpartum participated in the study. Each mother completed in random order three questionnaires: The PDSS, EPDS, and The Beck Depression Inventory-II (BDI-II). Immediately after completing these three questionnaires, each woman was interviewed by a nurse psychotherapist using the Structural Clinical Interview for DSM-IV Axis 1 Disorders. RESULTS: Twelve percent (n = 18) of the mothers were diagnosed with major postpartum depression, 19% (n = 28) with minor postpartum depression, and 69% (n = 104) with no depression. The PDSS was strongly correlated with both the BDI-II (r = 0.81) and the EPDS (r = 0.79). The ability of the PDSS to explain variance in diagnostic classification of postpartum depression above that explained by the BDI-II and EPDS (i.e., incremental validity) was assessed using hierarchical regression. After explaining variance in group classification by the other two depression instruments, the PDSS explained an additional 9% of the variance in depression diagnosis. Using receiver operating characteristic (ROC) curves, a PDSS cut-off score of 80 (sensitivity 94% and specificity 98%) is recommended for major postpartum depression and a cut-off score of 60 (sensitivity 91% and specificity 72%) for major or minor depression. CONCLUSION: Based on the results of this psychometric testing, the PDSS is considered ready for use in routine screening of mothers. PMID- 11393638 TI - Reliability of homeless women's reports: concordance between hair assay and self report of cocaine use. AB - BACKGROUND: To assess the concordance of homeless women's self-reported drug use with objective data. OBJECTIVE: To determine whether objective data (e.g., hair assays) are necessary supplements to self reports in assessing homeless women's cocaine use. METHOD: Self reports of cocaine use by 1,037 homeless women were compared to objective data based on radioimmunoassay of hair; independent correlates of cocaine use and underreporting were assessed using logistic regression. RESULTS: Forty-two percent of the women self-reported cocaine use in the past 6 months, whereas 49% had positive hair assays. Over 25% underreported cocaine use; however, underreporting decreased as hair cocaine levels increased. Predictors of underreporting included being Latino, younger and living primarily in shelters. Nevertheless, independent predictors of self-reported cocaine use and positive hair assays were identical. CONCLUSION: Homeless women's self reports of cocaine use are fairly accurate. Objective data are particularly critical for assessing cocaine use among subgroups who are fearful of sanctions or use cocaine relatively infrequently or in smaller amounts. PMID- 11393639 TI - Differences in plasma and nipple aspirate carotenoid by lactation status. AB - BACKGROUND: Dietary antioxidants, such as provitamin A carotenoid, have a protective effect against breast cancer. The transport of carotenoid from the blood into the breast microenvironment may be enhanced by lactation. OBJECTIVE: To examine the association between plasma and nipple aspirate carotenoid levels by lactation and post-wean status. METHODS: The sample consisted of 43 women, ages 18-45, who were at least 12 months postpartum. Women who had breastfed their last infant were at least 3 months post-wean. Women collected breast fluid every other day for 17 days and had a venipuncture for total nipple aspirate and plasma carotenoid, and completed a written health assessment. RESULTS: The association between plasma and nipple aspirate carotenoid levels was significant for breastfeeding women (r =.39, p=.03), but not for non-breastfeeding women (r =.31, p =.27). However, while the association between plasma and nipple aspirate carotenoid levels was significant for women at or less than 9 months post-wean (r =.65, p = .01), the effect for women after 9 months post-wean (r = .21, p =.45) was not significant. CONCLUSION: Lactation may be protective by enhancing the delivery of chemopreventive substances available in the blood to the cell level of the breast, even after breast involution has occurred post lactation. PMID- 11393640 TI - Selenium and glutathione peroxidase with beta-thalassemia major. AB - BACKGROUND: Chronic iron-overload is a major cause of organ failure and mortality worldwide, but its pathogenesis remains to be elucidated. OBJECTIVES: To examine the relationship between various measures of body iron burden, selenium concentrations and glutathione peroxidase (GPx) activity in patients with beta thalassemia major. METHODS: An age- and gender-matched case control study was conducted to examine the relationship between various measures of body iron burden (serum ferritin, transferrin saturation, total serum iron), plasma concentrations of selenium and glutathione peroxidase (GPx) activity in patients with homozygous beta-thalassemia major (N = 20) and healthy controls (N = 10). Ten patients received the experimental oral chelator L1 and ten received chelation therapy with subcutaneous desferal. RESULTS: Significantly decreased plasma concentrations of selenium (microg/L) were observed in patients chelated with L1 (1.4 +/- 0.2) or desferal (1.4 +/- 0.1), in comparison to healthy controls (1.8 +/- 0.1, p < 0.01). Significantly decreased plasma activity of GPx (microg/L) was observed in patients chelated with L1 (166 +/- 43) or desferal (178 +/- 46), in comparison to healthy controls (296 +/- 22, p < 0.001). Significantly increased concentrations of all measures of body iron burden were observed in beta-thalassemia patients, in comparison to healthy controls (p < 0.001). CONCLUSION: Patients with beta-thalassemia major and chronic iron overload have decreased concentrations of the essential element selenium and the protective selenium-dependent antioxidant enzyme GPx. Additional research examining the effects of dietary antioxidant supplementation with selenium on these aforementioned parameters in patients with beta-thalassemia major and iron overload is warranted. PMID- 11393641 TI - Methodological considerations in children's focus groups. AB - Focus groups are a well-known qualitative approach to gathering data in health science research. The literature on focus groups, however, primarily discusses adults as subjects. Unfortunately, the scant reports of studies using children as participants in focus groups have not described their methods in detail. This article discusses the use of children (age 6-12) in focus groups, and highlights methodological considerations in this approach, with particular attention to the integration of developmental principles. Focus groups with children can capture their perspectives, original ideas, and insights, which are often neglected in more traditional pediatric research. Focus groups can also serve as an innovative approach to understanding children's experiences from a developmental perspective. Further, focus groups free children and investigator from the data gathering limitations placed by literacy/reading levels that plague quantitative methods using self-report. By using relatively homogeneous groups, common cultural, emotional, and cognitive processes and responses are revealed that normally would not come to light in structured data collection. Focus groups offer a rich, interactive and developmentally effective approach to planning, content and evaluation in research with children. PMID- 11393642 TI - Methodological considerations for grounded theory research in critical care settings. AB - BACKGROUND: The context of critical care and human phenomena involved in critical illness offer rich opportunity for nursing research. Naturalistic investigation with grounded theory methods can uncover previously unexamined elements and interactions in the critical care setting. This article presents methodological considerations for conducting grounded theory research in fast-paced physiologically and technologically complex critical care settings. APPROACH: Critique and recommendations are based in review and analysis of grounded theory research in adult critical care settings and on the literature describing grounded theory methods. The authors' experiences in medical and surgical intensive care units provide added practical context for this article. RESULTS: Barriers to achieving grounded theory in critical care settings, such as communication impairments, participant attrition, and observational difficulties, are explored. Methodological strategies and data sources particular to critical care settings are also discussed. Critical care settings offer a variety of data sources that should be rigorously pursued in grounded theory research. DISCUSSION: Given current trends in healthcare treatments and demographics, future research must examine physiological and technological data as integral components of basic social psychological or social structural processes in critical care interactions and should include technology as a component of nurse patient interaction. PMID- 11393643 TI - Vagal tone, development, and Gray's motivational theory: toward an integrated model of autonomic nervous system functioning in psychopathology. AB - In the last decade, cardiac vagal tone has emerged as a psychophysiological marker of many aspects of behavioral functioning in both children and adults. Research efforts during this time have produced an extensive list of vagal tone correlates that includes temperamental variables as well as both anxious/internalizing and disruptive/externalizing behaviors. This potentially confusing state of affairs is compounded by developmental shifts in vagal tone behavior relations that to date have not been elucidated. In this paper, the vagal tone literature is reviewed, and discrepancies, including the lack of specificity of vagal tone as a psychophysiological marker, are clarified. Such clarification requires that we (a) view vagal tone-behavior relations in developmental context, (b) juxtapose vagal tone-behavior relations in typical and atypical samples, and (c) consider the parasympathetic underpinnings of vagal tone as but one component in a broader model of autonomic nervous system functioning. Such a model is provided by combining Gray's motivational theory with Porges's polyvagal theory. Together these models account for behavioral and emotional differences in a diverse range of psychological disorders that are not differentiated by either model alone. Moreover, use of the integrated model offers a theory-driven approach to the study of autonomic nervous system-behavior relations. PMID- 11393644 TI - Attachment, parenting, and marital dissatisfaction as predictors of disruptive behavior in preschoolers. AB - The aim of this study was to examine if an insecure coercive attachment pattern is associated with disruptive behavior in preschoolers, as well as to examine the concurrent and joint effects of attachment pattern, marital dissatisfaction, and ineffective parenting practices on disruptive behavior. Participants included 60 preschoolers and their mothers, recruited from three sites to ensure an adequate range of disruptive behavior. The Preschool Assessment of Attachment (Crittenden, 1992) was used to measure attachment pattern. Results of an analysis of variance revealed that children in the coercively attached dyads scored significantly higher on the measure of disruptive behavior than either the defended or secure children. Results of a hierarchical regression analysis indicated that the combination of a coercive pattern of attachment, marital dissatisfaction, and permissive parenting practices accounted for a significant proportion of the variance in disruptive behavior in preschoolers. These data suggest that a specific type of insecure attachment, a coercive pattern, is associated with disruptive behavior in preschoolers. Also, the data are consistent with previous findings of associations among marital dissatisfaction, ineffective parenting practices, and disruptive behavior. PMID- 11393645 TI - The development of dissociation in maltreated preschool-aged children. AB - Dissociation reflects disruptions in the integration of memories, perception, and identity into a coherent sense of self, and may develop following childhood maltreatment. The preschool years were identified as an important period for the development of dissociation. However, prior research has not examined the development of dissociation during this time. In order to address this gap, evidence of dissociation in 45 maltreated children, assessed for sexual abuse, physical abuse, and neglect, was compared with dissociation in 33 nonmaltreated children. Rather than depend on adult observer reports of behavior, the study sought to gain an understanding of dissociation from the child's own point of view. Because self-reports have limitations with such young children, a measure of dissociation evidenced in children's narrative story-stem completions was utilized. Maltreated children, especially physically abused children and sexually abused children, demonstrated more dissociation than did nonmaltreated children. Moreover, during the preschool period maltreated and nonmaltreated children followed different trajectories such that dissociation increased for maltreated children but did not do so for nonmaltreated children. Findings suggest that although the self is normatively integrated during the preschool period, it becomes increasingly fragmented for some maltreated children. Results are discussed in terms of cascading effects of maltreatment throughout development, and the importance of developmentally sensitive interventions. PMID- 11393646 TI - Correlates of clinic referral for early conduct problems: variable- and person oriented approaches. AB - The current study utilized both variable- and person-oriented analyses to examine correlates of early disruptive behavior problems. Participants included 80 preschool boys referred to a child psychiatry clinic and diagnosed with oppositional defiant disorder (with or without attention-deficit hyperactivity disorder) and 80 case-matched normal comparison boys. The study examined four domains of correlates: vulnerable child characteristics, poor parenting practices, insecure attachment, and adverse family ecology. Results indicated that the combination of these factors provided relatively high sensitivity (81%) and specificity (85%), clearly differentiating referred from comparison boys. A dramatic increase in clinic status occurred when three or more factors were present, and specific combinations of factors were differentially predictive of conduct problems. However, no correlates were found to be either necessary or sufficient for clinic status. By maintaining the integrity of individual cases, person-oriented analyses were able to answer different questions than more traditional variable-oriented analyses. Discussion focuses on the value of person oriented analyses for understanding heterogeneous clinical groups. PMID- 11393647 TI - Structure and variability in the developmental trajectory of children's externalizing problems: impact of infant attachment, maternal depressive symptomatology, and child sex. AB - This study explores the developmental trajectory of externalizing problems in a sample of 101 children of adolescent mothers from preschool through third grade using hierarchical linear models (HLM). First, a detailed assessment of the structure of the developmental trajectory of externalizing problems is provided. Second, the impact of three risk factors (infant attachment, maternal depressive symptomatology, and child sex) on the developmental course of externalizing problems is assessed. Both avoidant and disorganized attachment and higher levels of maternal depressive symptomatology were associated with higher levels of externalizing problems at 9 years of age. Girls also showed higher externalizing problems relative to their same-sex peers than did boys. In addition, maternal depressive symptomatology related to the rate of change in these problems over time: the greater the mother's depression, the faster externalizing problems tended to increase. Although the overall level of maternal depressive symptomatology was related to children's externalizing problems for secure, avoidant, and disorganized groups, changes in maternal depressive symptomatology over time predicted levels of externalizing problems only for children with avoidant insecure attachments. PMID- 11393648 TI - Effects of grade retention on academic performance and behavioral development. AB - This study examined the controversial practice of grade retention and children's academic and behavioral adjustment using data from the Quebec Longitudinal Study of Kindergarten Children. We employed an autoregressive modeling technique to detect the impact of being held back during primary school on subsequent academic performance and behavioral development until age 12 years. The results indicate both a short- and long-term negative influence on academic performance for boys and girls. Children's anxious, inattentive, and disruptive behaviors persisted and, in some cases, worsened after grade retention. These prospective associations were long lasting and more pronounced when grade retention occurred early in primary school. Boys were more vulnerable to the negative influence of grade retention on academic performance and classroom disruptiveness. Disruptive behavior in girls was comparatively less associated with long-term consequences than boys. Nevertheless, girls experienced both short- and long-term academic performance problems in the aftermath of grade retention. Children's prosocial behavior appeared unaffected by grade retention. These results are independent of what would have been expected by the natural course of academic and behavioral development. PMID- 11393649 TI - Socialization of children's emotion regulation in mother-child dyads: a developmental psychopathology perspective. AB - This study investigated the socialization of children's emotion regulation in 25 physically maltreating and 25 nonmaltreating mother-child dyads. Maltreating mothers and their 6- to 12-year-old children were recruited from two parenting programs affiliated with Children's Protective Services with a control group matched on race, SES, child gender, and child age. Children and their mothers were interviewed individually about their (a) management of emotional expression. (b) strategies for coping with emotional arousal, and (c) anticipated consequences following emotional displays. Compared to controls, maltreated children expected less maternal support in response to their emotional displays, reported being less likely to display emotions to their mothers, and generated fewer effective coping strategies for anger. Maltreating mothers indicated less understanding of children's emotional displays and fewer effective strategies for helping children to cope with emotionally arousing situations than nonmaltreating mothers. Further, findings indicated that maternal socialization practices (e.g., providing support in response to children's emotional display, generating effective coping strategies for their child) mediate the relation between child maltreatment and children's regulation of emotional expression and emotional arousal. These findings suggest that children's emotion regulation strategies are influenced by their relationship with their social environment (e.g.. physically maltreating, nonmaltreating) and that the experience of a physically maltreating relationship may interfere with children's emotional development. PMID- 11393650 TI - Peer rejection in childhood, involvement with antisocial peers in early adolescence, and the development of externalizing behavior problems. AB - A longitudinal, prospective design was used to examine the roles of peer rejection in middle childhood and antisocial peer involvement in early adolescence in the development of adolescent externalizing behavior problems. Both early starter and late starter pathways were considered. Classroom sociometric interviews from ages 6 through 9 years, adolescent reports of peers' behavior at age 13 years, and parent, teacher, and adolescent self-reports of externalizing behavior problems from age 5 through 14 years were available for 400 adolescents. Results indicate that experiencing peer rejection in elementary school and greater involvement with antisocial peers in early adolescence are correlated but that these peer relationship experiences may represent two different pathways to adolescent externalizing behavior problems. Peer rejection experiences, but not involvement with antisocial peers. predict later externalizing behavior problems when controlling for stability in externalizing behavior. Externalizing problems were most common when rejection was experienced repeatedly. Early externalizing problems did not appear to moderate the relation between peer rejection and later problem behavior. Discussion highlights multiple pathways connecting externalizing behavior problems from early childhood through adolescence with peer relationship experiences in middle childhood and early adolescence. PMID- 11393651 TI - Childhood predictors differentiate life-course persistent and adolescence-limited antisocial pathways among males and females. AB - This article reports a comparison on childhood risk factors of males and females exhibiting childhood-onset and adolescent-onset antisocial behavior, using data from the Dunedin longitudinal study. Childhood-onset delinquents had childhoods of inadequate parenting, neurocognitive problems, and temperament and behavior problems, whereas adolescent-onset delinquents did not have these pathological backgrounds. Sex comparisons showed a male-to-female ratio of 10:1 for childhood onset delinquency but a sex ratio of only 1.5:1 for adolescence-onset delinquency. Showing the same pattern as males, childhood-onset females had high risk backgrounds but adolescent-onset females did not. These findings are consistent with core predictions from the taxonomic theory of life-course persistent and adolescence-limited antisocial behavior. PMID- 11393653 TI - Object relations as a predictor of adjustment in conjugal bereavement. AB - The impact of object relations on adjustment in conjugal bereavement was examined. At approximately 6 months postbereavement, 46 midlife bereaved participants engaged in a narrative interview in which they were asked to discuss their past relationship with their deceased spouse. The Westen et al. object relations scoring system was applied to these narratives. Participants also completed depression and grief-specific symptom measures at 6 months and again at 14 and 25 months postbereavement. Object relations correlated differently with grief-specific symptoms and depression: it was more strongly negatively associated with 6 month postloss grief-specific symptoms while more strongly negatively correlated with depression at 25 month postloss. In a growth curve analysis, more mature object relations was also predictive of a faster rate of decrease in depression over time. The results were discussed in terms of current theoretical perspectives on what is required in successful adaptation to conjugal bereavement. PMID- 11393652 TI - Why are children born to teen mothers at risk for adverse outcomes in young adulthood? Results from a 20-year longitudinal study. AB - This 20-year longitudinal study showed that the young adult offspring of teen mothers are at risk for a range of adverse outcomes including early school leaving, unemployment, early parenthood, and violent offending. We tested how much the effect of teen childbearing on offspring outcomes could be accounted for by social selection (in which a woman's characteristics that make her an inadequate parent also make her likely to bear children in her teens) versus social influence (in which the consequences of becoming a teen mother also bring harm to her children, apart from any characteristics of her own). The results provided support for both mechanisms. Across outcomes, maternal characteristics and family circumstances together accounted for approximately 39% of the effect of teen childbearing on offspring outcomes. Consistent with a social-selection hypothesis, maternal characteristics accounted for approximately 18% of the effect of teen childbearing on offspring outcomes; consistent with a social influence hypothesis, family circumstances accounted for 21% of the teen childbearing effect after controlling for maternal characteristics. These results suggest that public policy initiatives should be targeted not only at delaying childbearing in the population but at supporting individual at-risk mothers and their children. PMID- 11393654 TI - Allele-specific binding of the ubiquitous transcription factor OCT-1 to the functional single nucleotide polymorphism (SNP) sites in the tumor necrosis factor-alpha gene (TNFA) promoter. AB - The tumor necrosis factor-alpha (TNFA) is characterized by several single nucleotide polymorphisms (SNPs) in its promoter region. Interestingly some of these SNPs appear to influence TNFA expression and susceptibility to various human diseases, but the molecular mechanisms by which such possibly functional SNPs modulate TNFA expression are poorly understood. In this study we show allele specific binding of the ubiquitous transcription factor OCT-1 to the SNP sites at positions -863 and -857 in the promoter, which appear to affect TNFA expression: the protein was associated with variant allele possessing either -863A or -857T, but rarely with the common allele (-863C and -857C). The evidence presented here, therefore, suggests the possibility that OCT-1 could contribute to the modulation of TNFA expression by means of its allele-specific binding manner. PMID- 11393655 TI - Organization of the chemokine genes in the human and mouse major clusters of CC and CXC chemokines: diversification between the two species. AB - Chemokines are a family of small cytokines that play essential roles in the directed migration of various types of leukocytes. Based on the arrangement of the conserved cysteine residues, they are classified into two major subfamilies, CXC and CC, and two minor subfamilies, C and CX3C. So far, more than 40 members of this family have been identified in humans. Strikingly, the majority of CXC chemokine genes and that of CC chemokine genes are closely clustered at chromosomes 4q12-21 and 17q11.2, respectively. Similarly, the mouse major CXC and CC chemokine gene clusters are located on chromosomes 5 and 11, respectively. In order to understand the evolutionary processes that generated large numbers of CXC and CC chemokine genes in the respective chromosomal sites, we have constructed BAC and YAC contigs covering the human and mouse major clusters of CXC and CC chemokine genes. The results reveal that the organizations of CXC and CC chemokine genes in the major clusters are quite diverged between the two species most probably due to very recent gene duplications and rearrangements. Our results provide an important insight into the evolutionary processes that generated the major chemokine gene clusters and also valuable information in assigning the orthologues between human and mouse major cluster chemokines. PMID- 11393656 TI - The CC chemokine receptor 5 delta32 mutation is not associated with inflammatory bowel disease (IBD) in NE England. AB - Inflammatory bowel disease (IBD) is a group of chronic inflammatory diseases of the gastrointestinal tract of unknown aetiology. Evidence of abnormalities in immune regulation and cytokine production in patients with IBD has led to investigations of various immuno-regulatory genes as potential candidate susceptibility loci. Studies using whole genome scanning have highlighted chromosomes 3, 7, 12 and 16. A 32 base-pair deletion in the CC-chemokine receptor 5 gene (CCR5-A32, chromosome 3p21.3) has been associated with susceptibility to IBD. We have investigated CCR5 as a candidate susceptibility gene in 350 patients (251 with ulcerative colitis and 99 with Crohn's disease) and 103 controls using polymerase chain reaction. There were no significant differences in the distribution of CCR5 genotypes or frequencies comparing patients and controls, or associations with extent of colitis. In contrast to preliminary data, these findings suggest no evidence for involvement of this mutation in susceptibility/resistance or disease progression in IBD. PMID- 11393657 TI - Two novel single-nucleotide polymorphisms of the Caspase-9 (CASP9) gene in the Japanese population. AB - We identified two single-nucleotide polymorphisms (SNPs) in the human Caspase-9 (CASP9) gene (1p36.3), which encodes an apoptosis-related cysteine protease, by screening all exons and exon-intron boundaries. These SNPs were present in coding regions. One of coding SNPs reflected an amino-acid substitution; an A to G transition at codon 221 in exon 5 would encode arginine instead of glutamine. As the gene is implicated in apoptotic cascade, the polymorphic sites will serve as useful markers for genetic studying of disorders affecting immune response and cancer susceptibility in humans. PMID- 11393658 TI - Cytokine gene polymorphism in human disease: on-line databases, supplement 1. PMID- 11393659 TI - A full genome screening in a large Tunisian family affected with thyroid autoimmune disorders. AB - The autoimmune thyroid diseases (AITDs) including Graves' disease (GD) and Hashimoto's thyroiditis (HT) are inherited as complex traits. We initiated a whole genome linkage study of patients with AITD, in order to identify the susceptibility genes involved in their pathogenesis. We studied 39 patients affected with GD or HT and 68 related controls, who belonged to a large consanguinous family composed of more than 200 members. Linkage analysis was performed using the lod score method under two arbitrary models, one dominant and one recessive. A positive lod score was found for D2S171, assuming a recessive mode of inheritance and 50% penetrance, which suggests the presence of a major AITD susceptibility gene on chromosome 2p21. However, no linkage was found with microsatellite markers spanning the HLA system. This locus localised outside MHC will be of interest for investigation of other autoimmune disorders. PMID- 11393660 TI - Class II HLA associations with autoantibodies in scleroderma: a highly significant role for HLA-DP. AB - Scleroderma is a condition of variable phenotype characterised by fibrosis of the skin and internal organs. There is a range of disease-specific autoantibodies found in the sera of patients. The aims of this study were to: (1) investigate the role of the MHC and particularly HLA-DP in the production of autoantibodies; (2) investigate clinical associations with autoantibodies. We have performed HLA class II typing using PCR with sequence-specific primers on DNA samples from 202 scleroderma patients and 307 UK control subjects. All patients had well defined clinical phenotypes. Sera from patients were examined for the presence of disease specific autoantibodies in particular the anti-topoisomerase autoantibody (ATA), the anti-centromere autoantibody (ACA) and the anti-RNA polymerase autoantibody (ARA). There was a striking association between HLA-DPB1*1301 and ATA (Pcorr = 0.0001). In addition, ATA was associated with HLA-DRB1*11 and the anticentromere autoantibody (ACA) with HLA-DRB1*04, HLA-DRB1*08 (P = 0.001) and HLA-DQB1 alleles with a glycine residue at position 26. Very strong associations were detected between clinical phenotypes and autoantibodies. ATA was associated with pulmonary fibrosis (P = 0.00002), anti-RNA polymerase autoantibody (ARA) with renal involvement (P = 0.0000006) and diffuse skin disease (P = 0.00001), and ACA with limited skin involvement (P = 0.00002) and protection against pulmonary fibrosis (P = 0.0000003). We have identified a significant association between the ATA and HLA-DPB1*1301 which may provide an insight into how this autoantibody is formed. Patient clinical characteristics depend on the autoantibodies they carry. PMID- 11393661 TI - Tumour necrosis factor 5' promoter single nucleotide polymorphisms influence susceptibility to rheumatoid arthritis (RA) in immunogenetically defined multiplex RA families. AB - Tumour necrosis factor (TNF) plays a pivotal role in the pathogenesis of rheumatoid arthritis (RA) and it has been shown that the TNF-lymphotoxin (TNF-LT) region influences susceptibility to RA. To investigate the role of the TNF-LT locus further, inheritance of TNF 5' promoter alleles was determined in multiplex RA families. Six previously defined TNF promoter single nucleotide polymorphisms (SNPs) (-238, -308, -376, -857, -863, -1031) were observed in these families and in addition, a heretofore undocumented adenine (A) to cytosine (C) substitution at position -572 relative to the transcription start site was defined. TNF 5' promoter SNPs were found to co-segregate with specific TNF microsatellite haplotypes. In particular, the SNP -308A allele was found to be inherited with the TNF a2, b3, c1, d1, e3 (H2) microsatellite haplotype (P < 0.001) which had previously been found to be associated with RA in individuals heterozygous for the HLA-DR 'shared epitope' (SE). When the data were stratified by the presence of the SE with further stratification according to SE DR subtypes and analysed by transmission disequilibrium test (TDT) for which offspring were assumed independent, the -308A and -857T alleles were found to be associated with RA in patients carrying the SE (P = 0.0076 and 0.0063 respectively). The data were further stratified to analyse for association in individuals homozygous or heterozygous for SE alleles. Results showed that the -308A allele was significantly associated with RA susceptibility in individuals heterozygous for the SE (P < 0.001) with the significance only occurring in patients carrying HLA DR4 (P < 0.001), while the -857T allele was significant in individuals homozygous for the SE (P = 0.0039). Further analysis using the pedigree disequilibrium test (PDT) which conservatively adjusts for all sources of familial correlation except that conferred by linkage disequilibrium still indicated a significant role for the -308A and -857T alleles. These data provide evidence that TNF promoter SNPs may play an independent role in RA susceptibility in specific immunogenetically defined groups of RA patients. PMID- 11393662 TI - Profiling of differential gene expression in activated, allergen-specific human Th2 cells. AB - Th2 cells play a pivotal role in the pathogenesis of allergic diseases, including asthma, but the molecular basis of the Th1/Th2 dichotomy and the precise molecular pathways leading to Th2-dominant immune responses are still unclear. To this end, we have combined suppression subtractive hybridization (SSH) and high throughput analysis of cDNA arrays spotted with IMAGE clones to determine the profile of differential gene expression in human allergen-specific Th2 cells. Allergen-stimulated Th2 cells were used as the tester, and either resting Th2 cells or stimulated Th1 cells were used as the driver. SSH was used to equalize different mRNA levels and remove common sequences between the tester and the driver. Comparison of cDNA arrays probed with subtracted tester and non subtracted driver provided a profile of Th2-selective gene expression. Analysis of 77 sequence-confirmed and differentially expressed genes in Th2 cells showed predominant EST sequences, representing 80% of sequences analyzed. The pattern of gene expression in 19 selected sequences was further analyzed in additional Th1 and Th2 clones. A total of 15 sequences showed predominant expression in Th2 cells, while the remaining four EST sequences showed no detectable amplification signal. The database containing Th2-selective genes will further our understanding of Th2 cell function and the genetic basis of allergic diseases. PMID- 11393663 TI - Mannose-binding lectin gene: polymorphisms in Japanese patients with systemic lupus erythematosus, rheumatoid arthritis and Sjogren's syndrome. AB - Mannose-binding lectin (MBL) is a key element of the innate immunity, with a structure similar to complement C1q. Serum MBL levels are greatly affected by the polymorphisms of the MBL gene. In particular, codon 54 mutation of the MBL gene results in a significant reduction of serum MBL. To determine whether polymorphism of the MBL gene is associated with occurrence of systemic lupus erythematosus (SLE), rheumatoid arthritis and Sjogren's syndrome in the Japanese population, we analyzed the MBL gene polymophisms of these patients and controls, by polymerase chain reaction-restriction fragment length polymorphism methods. We found that patients studied had a significantly higher frequency of having homozygous codon 54 mutation compared to controls. In particular, patients with SLE or Sjogren's syndrome showed higher probabilities of being homozygous for this mutation. Among subjects with the same genotype, SLE patients tended to have higher serum MBL concentration than controls. Analysis of the promotor region suggested that SLE patients heterozygous for the codon 54 mutation have a higher probability of having a low producing haplotype for the gene without the codon 54 mutation. We conclude that persons homozygous for codon 54 mutation of the MBL gene may be prone to occurrence of autoimmune disorders including SLE, in the Japanese. MBL may have protective effects on occurrence and progression of SLE. PMID- 11393664 TI - Aldosterone and the hypertensive kidney: its emerging role as a mediator of progressive renal dysfunction: a paradigm shift. AB - End-stage renal disease (ESRD) comprises an enormous public health burden, with an increasing incidence and prevalence. Hypertension is a major risk factor for progressive renal disease. This escalating prevalence suggests that newer therapeutic interventions and strategies are needed to complement current antihypertensive approaches. Although much evidence demonstrates that angiotensin II mediates progressive renal disease, recent evidence also implicates aldosterone as an important pathogenetic factor in progressive renal disease. Several lines of experimental evidence demonstrate that selective blockade of aldosterone, independent of renin-angiotensin blockade, reduces proteinuria and nephrosclerosis in the spontaneously hypertensive stroke-prone rat model and reduces proteinuria and glomerulosclerosis in the subtotally nephrectomized rat model (i.e. remnant kidney). Whereas pharmacological blockade with angiotensin II receptor blockers and angiotensin-converting enzyme inhibitors reduces proteinuria and nephrosclerosis/ glomerulosclerosis, selective reinfusion of aldosterone restores these abnormalities despite continued renin-angiotensin blockade. Aldosterone may promote fibrosis by several mechanisms, including plasminogen activator inhibitor-1 expression and consequent alterations of vascular fibrinolysis, by stimulation of transforming growth factor-beta 1, and by stimulation of reactive oxygen species. Based on this theoretical construct, randomized clinical studies will be initiated to delineate the potential renal protective effects of antihypertensive therapy utilizing aldosterone receptor blockade. PMID- 11393665 TI - Impact of the Gubbio population study on community control of blood pressure and hypertension. Gubbio Study Research Group. AB - BACKGROUND: Awareness and treatment of hypertension have markedly increased in the last 30 years in most parts of the world, but a satisfactory control of blood pressure is still infrequent OBJECTIVES: To describe trends in community control of hypertension and blood pressure levels in the small town of Gubbio, Italy. METHODS: Large samples of the populations (aged 30-79 years) were examined 6 years apart for measurement of blood pressure, other cardiovascular risk factors and knowledge, attitude and practice towards control of hypertension. Data were available from a total of 1125 men and 1445 women with two examinations and 1566 men and 1658 women with at least one examination. Two different definitions of hypertension were used (old definition: systolic blood pressure (SBP) > or = 160 mmHg or diastolic blood pressure (DBP) > or = 95 mmHg or use of antihypertensive drugs; recent definition: SBP > or = 140 mmHg or DBP > or =90 mmHg or use of anti hypertensive drugs). RESULTS: Awareness, treatment and control of hypertension increased from one survey to the other, whatever definition of hypertension was used. Control rose from 41 to 63% (old definition) and from 12 to 24% (recent definition). In a 6-year period, the average population SBP declined 2-8 mmHg and DBP declined 2-3 mmHg depending on type of analysis, against an expected rise of 9 mmHg for SBP and 2 mmHg for DBP. These trends are partly explained by a marked decline in alcohol consumption and by more common and intensive anti-hypertensive treatment, while change in body mass index, which showed a slight but systematic increase, cannot be considered as a contributor to this trend. CONCLUSIONS: An epidemiological study has motivated a population group and its medical profession towards a better control of hypertension. PMID- 11393666 TI - Alcohol consumption and risk for hypertension in middle-aged Japanese men. AB - OBJECTIVE: To investigate the association of alcohol intake with development of hypertension. DESIGN: Longitudinal study (followed from 1990 to 1999). SETTING: Work site in Osaka, Japan. PARTICIPANTS: Japanese male office workers (n = 1,310) hypertension-free, with systolic blood pressure (SBP) less than 140 mmHg, diastolic blood pressure (DBP) less than 90 mmHg, no medication for hypertension, and no past history of hypertension, 30 to 59 years of age at study entry. MAIN OUTCOME MEASURE: Incidence of hypertension. RESULTS: After controlling for potential predictors of hypertension, the relative risk for hypertension (SBP 140 mmHg or greater and/or DBP 90 mmHg or greater or receipt of antihypertensive medication) compared with non-drinkers was 1.52 [95% confidence interval (CI), 1.05 to 2.19] for persons who drank 0.1 to 22.9 g/day of ethanol, 1.81 (95% CI, 1.29 to 2.54) for those who drank 23.0 to 45.9 g/day of ethanol, 2.12 (95% CI, 1.53 to 2.94) for those who drank 46.0 to 68.9 g/day of ethanol, and 2.48 (95% CI, 1.75 to 3.52) for those who drank > or = 69.0 g/day of ethanol (for trend, P < 0.001). The relative risk for hypertension in current drinkers versus non drinkers was stronger among men with a body mass index (BMI) less than 24.2 kg/m2 than among men with a BMI 24.2 kg/m2 or more, although the absolute risk was greater in more obese men. CONCLUSIONS: The risk for hypertension increased in a dose-dependent manner as alcohol intake increased in middle-aged Japanese men. The increased relative risk for hypertension associated with alcohol was more pronounced in leaner men. PMID- 11393667 TI - Is C-reactive protein an independent risk factor for essential hypertension? AB - CONTEXT: C-reactive protein (CRP), predicts coronary heart disease incidence in healthy subjects and has been associated with decreased endothelium-dependent relaxation, a potential risk factor for hypertension. However, the relationship between CRP and hypertension has not been studied. OBJECTIVE: To assess whether circulating levels of CRP are independently related to essential hypertension. DESIGN: Cross-sectional population survey. We measured circulating levels of CRP, blood pressure and cardiovascular risk factors among participants. Binomial regression was used to calculate the adjusted effect of CRP on the prevalence of hypertension. SETTING: General community of Bucaramanga, Colombia. PARTICIPANTS: A random sample of 300 subjects > or = 30 years old. MAIN OUTCOME MEASURE: Arterial blood pressure. RESULTS: Overall hypertension prevalence was 46.0%. The unadjusted prevalence of hypertension was 58.7% in the highest quartile of CRP, but only 34.7% in the lowest quartile. After adjustment for age, sex, body mass index, family history of hypertension, fasting glycemia, sedentary behaviour, and alcohol consumption, the prevalence of hypertension was 1.14 [95% confidence interval (CI), 0.82, 1.58; P= 0.442], 1.36 (95% CI, 0.99, 1.87; P= 0.057) and 1.56 (95% CI, 1.14, 2.13; P = 0.005) times higher in subjects in the second, third and fourth quartiles of CRP, as compared to subjects in the first quartile. CONCLUSIONS: Our results suggest, for the first time, that CRP level may be an independent risk factor for the development of hypertension. However, because of the cross-sectional nature of our study, this finding should be confirmed in prospective cohort studies, aimed at elucidating the role of CRP in the prediction, diagnosis and management of hypertension. PMID- 11393668 TI - Combined effects of heart rate and pulse pressure on cardiovascular mortality according to age. AB - OBJECTIVES: The aim of the study was to assess the combined effects of pulse pressure (PP) and heart rate (HR) on cardiovascular mortality in a large French population. DESIGN: The study population was composed of 125,513 men and 96,301 women aged 16-95 years who had a health check-up at the IPC Center between January 1978 and December 1988. Subjects taking antihypertensive treatment were excluded. Mortality was assessed for an 8-year period. HR and PP were classified into three groups. HR groups were: < 60, 60-79 and > or = 80 beats per minute (bpm). PP groups were: < 50, 50-64 and > or = 65 mmHg. RESULTS: In men, PP and HR were both positively associated with cardiovascular mortality risk. In women, mean arterial pressure (MAP) but not PP or HR was associated with cardiovascular mortality. In men, a combined elevation of PP and HR was associated with an important increase of cardiovascular mortality risk. The group with the highest PP and the highest HR had a 4.8-fold increase in cardiovascular mortality risk as compared to the reference group (PP < 50 mmHg and HR < 60 bpm). This effect was more pronounced in younger men (5.4-fold increase) than in older men (3.7-fold increase), as compared to the reference groups of the same age. In women, the combined effects of PP and HR on cardiovascular mortality were not significant. CONCLUSION: A combined elevation of the two components of pulsatile arterial stress is associated with an important increase in cardiovascular mortality in men, especially in younger men. In women, steady-state stress (evaluated primarily by MAP), but not pulsatile stress, is an important determinant of cardiovascular mortality. PMID- 11393669 TI - Aortic pulse wave velocity as a marker of cardiovascular disease in subjects over 70 years old. AB - BACKGROUND: Aortic pulse wave velocity (PWV) is a significant and independent predictor of cardiovascular diseases (CVD) in hypertensive subjects and in patients with end-stage renal disease, but its contribution to cardiovascular risk in subjects between 70 and 100 years old has never been tested. PATIENTS: A cohort of 124 subjects (mean age: 87 +/- 7 years) was studied in two geriatric departments in a Paris suburb. Together with sphygmomanometric blood pressure measurements, aortic PWV was measured using a validated automatic device. RESULTS: Blood pressure, heart rate and body mass index, but not age, explained 48% of the PWV variability in this cohort. Furthermore, PWV was the major factor predicting the presence of CVD. The adjusted odds ratio was 17.44 (95% confidence intervals: 2.52-120.55). Antihypertensive drug therapy and low plasma albumin level had only an additive role. Blood pressure, particularly pulse pressure, had no predictive value. CONCLUSION: In 70-100-year-old subjects, aortic PWV is a strong independent marker of CVD, a finding that remains to be to confirmed by long-term longitudinal studies. PMID- 11393670 TI - Homozygosity for angiotensinogen 235T variant increases the risk of myocardial infarction in patients with multi-vessel coronary artery disease. AB - OBJECTIVE: Molecular variants of the angiotensinogen (AGT) and the angiotensin II type 1 receptor (ATR) genes have been associated with the risk of coronary artery disease (CAD) and myocardial infarction (MI), but data so far available are conflicting. The primary object of the paper is to verify this possible association by a rigorous, angiographically controlled study in a large sample of patients with or without multi-vessel CAD. DESIGN: We designed a large case control study in Italian patients candidates for coronary artery bypass grafting, with angiographically documented multi-vessel CAD, compared to subjects with angiographically documented normal coronary arteries. METHODS AND RESULTS: AGT M235T and ATR A1166C gene polymorphisms were analysed in 699 subjects; 454 patients were candidates for coronary artery bypass grafting, having angiographically documented (mainly multi-vessel) CAD. An appropriate documentation of previous MI was obtained from 404/454 (89%, 247 with and 157 without MI). Subjects (n = 245) with angiographically documented normal coronary arteries, were included as control group (CAD-free group). CAD patients had a substantial burden of conventional risk factors as compared with controls free of coronary atherosclerosis. Age, gender, smoking habit and number of stenosed vessels were the only differences between patients with or without previous myocardial infarction, who were similarly exposed to the other conventional risk factors (including hypertension). AGT M235T and ATR A1166C allele and genotype frequencies were similar between CAD and CAD-free patients. In the CAD group, AGT 235T allele was found more frequently in subjects with a previous myocardial infarction (0.494 versus 0.414; P < or = 0.05). By logistic regression, homozygosity for AGT 235T variant appeared to confer 1.9-fold increased risk for MI in both the univariate and the multivariate (adjusted for age, gender, smoking habit and number of stenosed vessels) model. CONCLUSIONS: AGT 235 T homozygous patients with multivessel CAD have an increased risk of myocardial infarction as compared with subjects with clinically similar phenotype but different genotype. PMID- 11393672 TI - Cigarette smoke-induced endothelium dysfunction: role of superoxide anion. AB - OBJECTIVES: Cigarette smoking is strongly associated with coronary artery disease and atherosclerosis. While smoking has been shown to impair endothelium-dependent vasorelaxation, the mechanisms involved are not completely understood. We investigated the role of superoxide anion and vasoconstricting prostanoids in cigarette smoke induced endothelial dysfunction. METHODS: Endothelial function was assessed in rat aortic rings exposed to cigarette smoke-treated Krebs buffer, by measuring agonist stimulated endothelium-dependent vasorelaxation. Treatment with superoxide dismutase (SOD) as well as ifetroban, thromboxane A2/prostaglandin endoperoxide H2 (TxA2/PGH2) receptor blocker and indomethacin (cyclooxygenase inhibitor) was used to investigate the role of superoxide anion and vasoconstricting eicosanoids on cigarette smoke-induced endothelial dysfunction. The effect of cigarette smoke on endothelial nitric oxide synthase (eNOS) catalytic activity was measured by conversion of L-arginine to L citrulline in rat aortas and rat endothelial cell homogenates supplemented with eNOS cofactors. RESULTS: Relaxations to receptor-dependent agonists, acetylcholine and adenosine diphosphate (ADP), as well as to a receptor independent agonist, A23187 (Ca2+ ionophore) were significantly impaired by cigarette smoke. Cigarette smoke did not impair relaxations to sodium nitroprusside, indicating preserved guanylate cyclase activity. Further, cigarette smoke did not affect eNOS catalytic activity in homogenates from either endothelial cells or aortas previously exposed to cigarette-smoketreated Krebs buffer. Treatment with SOD or ifetroban and in a lesser degree by indomethacin prevented cigarette-smoke-induced endothelial dysfunction. CONCLUSIONS: Taken together, our results suggest that cigarette smoking causes an increase in vascular superoxide production which results in decreased nitric oxide (NO) bioactivity and concomitantly increases production of cyclooxygenase dependent and independent vasoconstricting eicosanoids. PMID- 11393671 TI - Diagnosis of Liddle syndrome by genetic analysis of beta and gamma subunits of epithelial sodium channel--a report of five affected family members. AB - OBJECTIVE: To screen the gene mutation in beta and gamma subunits of the epithelial sodium channel (ENaC) of a Chinese family, some of whose members are clinically diagnosed as suffering from Liddle syndrome. METHODS: Twelve family members were recruited to the study. Among them, two brothers had been clinically diagnosed as suffering from Liddle syndrome. Peripheral blood samples were collected from all members of the family and total genomic DNA was prepared for genetic analysis. Polymerase chain reaction (PCR) was used for amplifying the last exon of beta (codon 513-673) and gamma (codon 503-632) subunits of the ENaC gene. PCR products were purified and subjected to a direct DNA sequence analysis. RESULTS: Genetic analysis of the beta ENaC gene revealed a missense mutation of CCC to CTC at codon 616 in four middle-aged men of the second generation and one young woman of the third generation. There was no mutation of the gamma ENaC gene in any of the individuals examined. CONCLUSION: Through direct DNA sequencing analysis, we diagnosed the disease present in five members of a Chinese family as Liddle syndrome, and excluded it in some other young offspring suffering from the monogenic disease. Our results provide further evidence that Pro616 is a critical amino acid that has a key role in the inhibition of sodium channel activity. PMID- 11393673 TI - Isradipine improves endothelium-dependent vasodilation in normotensive coronary artery disease patients with hypercholesterolemia. AB - OBJECTIVE: The dihydropyridine calcium antagonist isradipine has anti atherosclerotic effects in animals and improves endothelium-mediated nitric oxide (NO)-dependent vasodilation in vitro. As improved endothelial function may be beneficial we investigated its effects in patients with a high likelihood of endothelial dysfunction. DESIGN: Thirty patients (two female, age 55.4 +/- 10.5 years) with known coronary artery disease and elevated (> 6 mmol/l) total cholesterol (cholesterol: mean 6.7 +/- 0.78 mmol/l) or a cholesterol/high density lipoproteins (HDL) ratio of > 5 not on lipid lowering therapy, participated in the study. Endothelial vasodilator function was assessed before and after double blind, randomized administration of isradipine 5 mg/day or placebo for 3 months. METHODS: Endothelial function was assessed as forearm blood flow (FBF, venous occlusion plethysmography) responses to graded brachial artery infusions of acetylcholine (Ach), to the NO-synthase blocker NG-monomethyl-L-arginine (L-NMMA) and to the endothelium-independent vasodilator sodium nitroprusside (SNP). Blood pressure was measured either directly from the brachial arterial or by sphygmomanometer during clinic visits. RESULTS: Blood pressure was unchanged in both groups after 3 months (isradipine: 88.8 versus 92.1 mmHg; placebo: 81.0 versus 82.5 mmHg; NS) but cholesterol levels decreased similarly in both groups (isradipine: 6.7 versus 6.1 mmol/l, NS; placebo: 6.6 versus 5.9 mmol/l, P< 0.05). The vasodilator response to SNP and the decrease in FBF in response to blockade of NO synthesis by L-NMMA were unchanged in both groups. However, isradipine, but not placebo, enhanced the NO-dependent vasodilator response to Ach (P < 0.05). CONCLUSION: Isradipine improves acetylcholine-mediated vasodilation in hypercholesterolemic patients independent of changes in lipids or blood pressure. PMID- 11393674 TI - The nitric oxide synthase inhibitor L-NMMA potentiates noradrenaline-induced vasoconstriction: effects of the alpha2-receptor antagonist yohimbine. AB - OBJECTIVE: Alpha2-adrenoceptors can be found both on vascular smooth muscle cells and on the endothelium, where they exert opposing effects on vascular tone. In vitro, the stimulation of alpha2-adrenoceptors on endothelial cells leads to the release of vasodilating substances like nitric oxide (NO) and prostanoids. Little is known of this mechanism in vivo. DESIGN AND METHODS: We investigated the effects of the NO-synthase inhibitor L-NMMA (10(-6) mol) and the alpha2 adrenoceptor antagonist yohimbine (YO, 10(-10)-10(-6) mol) on noradrenaline (NA, 10(-12)-10(-8) mol)-induced vasoconstriction in the forearm skin microcirculation of 16 healthy volunteers using double injection technique and laser Doppler flowmetry. Results are expressed in perfusion units (PU) as differences from baseline and control in mean +/- SEM; the area under the time-response-curve was calculated (AUC). RESULTS: NA (10(-8)- 10(-12) mol) caused a marked, dose dependent reduction in blood flow (mean effect -745 +/- 84 AUC PU; P< 0.001 versus saline). NA-induced vasoconstriction was enhanced by L-NMMA (mean effect 916 +/- 72 AUC PU; P< 0.001 versus NA). YO (10(-6)-10(-10) mol) induced a significant, dose-dependent vasodilation (mean effect +/- 446 +/- 110 AUC PU; P < 0.05 versus control); high doses of YO (10(-6) mol) inhibited NA constriction (P < 0.001 versus NA), whereas lower doses of YO (10(-8)/10(-10) mol) had no effect or even increased NA-induced constriction. In the presence of L-NMMA, YO (10(-8) and 10(-10) mol) further potentiated NA-induced vasoconstriction (mean effect 1165 +/- 108 AUC PU; NS versus NA). CONCLUSION: These data demonstrate, that in humans in vivo, endogenous NO attenuates noradrenergic constriction. The effects of YO suggest that endothelial alpha2-adrenoceptors are involved in the release of NO and other vasodilating substances. Furthermore, there is an additive NO independent vasodilation, which can be unmasked by L-NMMA. PMID- 11393675 TI - Endothelial dysfunction in small resistance arteries of patients with non-insulin dependent diabetes mellitus. AB - OBJECTIVE: Arterial hypertension is frequently associated with the presence of endothelial dysfunction in human subcutaneous small resistance arteries, as evaluated by responses to acetylcholine or bradykinin; however it is not known whether patients with diabetes mellitus show similar alterations. Therefore, we have investigated endothelial function in subcutaneous arteries of normotensive subjects (NT), of patients with essential hypertension (EH), of patients with non insulin-dependent diabetes mellitus (NIDDM), as well as of patients with both essential hypertension and non-insulin-dependent diabetes mellitus (NIDDM+EH). PATIENTS AND METHODS: All subjects were submitted to a biopsy of the subcutaneous fat Small arteries were dissected and mounted on a micromyograph. The media to lumen ratio (M/L) was calculated. A concentration-response curve to acetylcholine, to bradykinin as well as to the endothelium-independent vasodilator sodium nitroprusside were performed. We also evaluated the contractile response to endothelin-1. Intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule 1 (VCAM-1) plasma levels were also measured. RESULTS: The vasodilatation to acetylcholine and bradykinin (but not to sodium nitroprusside) was significantly and similarly reduced in EH, in NIDDM, and in NIDDM+EH compared with NT. The contractile response to endothelin-1 was similarly reduced in EH, in NIDDM and in NIDDM+EH. Plasma ICAM-1 and VCAM-1 concentrations were higher in EH, NIDDM and NIDDM+EH than in NT. CONCLUSIONS: An evident endothelial dysfunction was detected in patients with NIDDM, and the simultaneous presence of EH did not seem to exert an additive effect. The contractile responses to endothelin-1 were reduced possibly as a consequence of ET(A) receptor down-regulation. PMID- 11393676 TI - Small artery remodeling is the most prevalent (earliest?) form of target organ damage in mild essential hypertension. AB - BACKGROUND: The heart and blood vessels are exposed to elevated blood pressure (BP) in hypertensive patients, but their changes in response to BP or non hemodynamic stimuli may be different, and occur with different time-courses. To evaluate this, we studied the prevalence of structural and functional alterations of resistance arteries and cardiac hypertrophy in patients with mild essential hypertension. METHODS: Resistance arteries were dissected from gluteal subcutaneous tissue from 38 hypertensive patients (47 +/- 1 years; 71% male; BP 148 +/- 2/99 +/- mmHg), studied on a pressurized myograph, and compared to those from 10 normotensives (44 +/- 3 years; 40% male; BP 113 +/- 4/76 +/- 2 mmHg). RESULTS: The prevalence of abnormal structure (media-to-lumen ratio, M/L) and impaired endothelial function (maximal acetylcholine response) was 97 and 58% (abnormal was defined as greater than mean + 1 SD of normotensives), or 63 and 34% (abnormal defined as greater than mean +/- 2SD). Thirty four percent of hypertensive patients exhibited left ventricular hypertrophy by echocardiography. When grouped into tertiles according to increasing ambulatory systolic BP (SBP), the highest BP tertile showed increased M/L (P< 0.01) and left ventricular mass index (LVMI, P < 0.05) and marginally decreased endothelial function (P= 0.07). LVMI was greatest in the tertile of patients with highest M/L (P< 0.05). Endothelial function was decreased in the tertile with greatest vascular stiffness (P< 0.01). By multivariate analysis, M/L correlated with ambulatory SBP (beta = 0.40, P= 0.02), and LVMI correlated with ambulatory SBP (beta = 0.41, P = 0.001) and body mass index (beta = 0.30, P< 0.05). Female sex influenced endothelial function negatively (beta = -0.63, P< 0.01). CONCLUSION: Structural alterations of resistance arteries were demonstrated in most hypertensive patients, followed by endothelial dysfunction and cardiac hypertrophy in a smaller number of hypertensives. Small artery structural remodeling may precede most clinically relevant manifestations of target organ damage in mild essential hypertension. PMID- 11393677 TI - Exercise training causes skeletal muscle venular growth and alters hemodynamic responses in spontaneously hypertensive rats. AB - OBJECTIVE: To investigate whether training changes skeletal muscle venular profile and hemodynamic responses to exercise we studied spontanesouly hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats submitted to training programme (T = 50-60% of VO2max). DESIGN: Training (T) was performed on a treadmill over a period of 13 weeks. Age-matched control groups were kept sedentary (S). T and S rats were chronically instrumented for hindlimb flow (HLF) and arterial pressure (AP) measurements at rest, during dynamic exercise and recovery in two different situations: control and after extensive intravenous blockade (hexamethonium + losartan + Nomega-nitro-L-arginine methyl ester + hydralazine). For morphometric analysis, skeletal muscle samples (gracilis) were obtained after transcardiac perfusion with fixative. RESULTS: T caused a significant reduction of resting mean arterial pressure (MAP) (-11%) only in the SHR group without changing basal HLF. In the sedentary SHR (SHRs), basal relative hindlimb resistance was increased by 45%, but was significantly reduced after T (P < 0.05). During dynamic exercise, MAP increased similarly (10-20 mmHg) in all groups. HLF increases were similar for the four groups up to 0.8 km/h; at higher workloads, HLF was higher in trained SHR (SHRT) versus trained WKY (WKYT) (3.9- versus 2.9-fold increase over basal HLF, respectively). After blockade (and pressure correction with IV phenylephrine infusion), steady-state exercise was performed with similar hindlimb vasodilation in all groups and was accompanied by MAP reduction (-17 +/- 8 mmHg) only in SHRT group. Skeletal muscle venular profile (density, diameter and lumen cross-sectional area) was similar in WKY(T), WKY(S) and SHR(S), but significantly increased in SHR(T). In this group the two fold increase in venule density was correlated with both the reduction in baseline MAP and the increase in HLF during dynamic exercise. CONCLUSIONS: The results suggest that increased venule density is a specific adaptation of SHR skeletal muscle to training. Venular growth may contribute to both the pressure lowering effect and the large HLF at high exercise intensities observed in the trained SHR. PMID- 11393678 TI - In-vitro and in-vivo inhibition of rat neutral endopeptidase and angiotensin converting enzyme with the vasopeptidase inhibitor gemopatrilat. AB - OBJECTIVES: Vasopeptidase inhibitors are single molecules that simultaneously inhibit neutral endopeptidase (NEP) and angiotensin converting enzyme (ACE). The aim of this study was to characterize in-vitro and in-vivo inhibition of NEP and ACE in the rat with the vasopeptidase inhibitor gemopatrilat. DESIGN AND METHODS: In-vitro NEP and ACE inhibition was studied by radioinhibitory binding assay using rat renal membranes and the specific NEP inhibitor radioligand 125I-RB104 and the specific ACE inhibitor radioligand 125I-MK351A, respectively (n = 3 per curve). In-vivo NEP and ACE inhibition was studied using in-vitro autoradiography in rats that received oral gemopatrilat (1, 3, 10 mg/kg; n = 4 per dose) and were killed 1 h later, or received oral gemopatrilat (3, 10 mg/kg) and were killed at time points 1, 2, 4, 8, 18, 24 and 48 h (n = 4 per time point). RESULTS: Gemopatrilat caused a concentration-dependent displacement of specific radioligands from renal membrane NEP (IC50 305 +/- 5.4 nmol/I) and ACE (IC50 3.6 +/- 0.02 nmol/). In the dose-response study gemopatrilat (1, 3 and 10 mg/kg) caused significant inhibition of plasma ACE (P< 0.01), and renal ACE and NEP (3, 10 mg/kg, P < 0.01). In the time course experiment, gemopatrilat (10 mg/kg) increased plasma renin activity for 8 h (P< 0.01) and inhibited plasma ACE (P< 0.05), renal NEP (P< 0.01) and renal ACE (P< 0.05) for 48 h. CONCLUSIONS: Gemopatrilat is a potent in-vitro vasopeptidase inhibitor that also causes prolonged inhibition of circulating and renal ACE and renal NEP after a single oral dose. The data suggest that gemopatrilat may be a useful addition to existing vasopeptidase inhibitors in the treatment of cardiovascular disease. PMID- 11393679 TI - Breathing patterns and cardiovascular autonomic modulation during hypoxia induced by simulated altitude. AB - OBJECTIVE: To assess the influence of different breathing patterns on autonomic cardiovascular modulation during acute exposure to altitude-induced hypoxia. DESIGN: We measured relative changes in minute ventilation (VE), oxygen saturation (%SaO2), spectral analysis of RR interval and blood pressure, and response to stimulation of carotid baroreceptors (neck suction) at baseline and after acute (1 h) hypobaric hypoxia (equivalent to 5,000 m, in a hypobaric chamber). METHODS: We studied 19 human subjects: nine controls and 10 Western yoga trainees of similar age, while breathing spontaneously, at 15 breaths/min (controlled breathing) and during 'complete yogic breathing' (slow diaphragmatic + thoracic breathing, approximately 5 breaths/min) in yoga trainees, or simple slow breathing in controls. RESULTS: At baseline %SaO2, VE and autonomic pattern were similar in both groups; simulated altitude increased VE in controls but not in yoga trainees; %SaO2 decreased in all subjects (P< 0.0001), but more in controls than in yoga trainees (17 versus 12%, 14 versus 9%, 14 versus 8%, all P< 0.05 or better, during spontaneous breathing, controlled breathing and yogic or slow breathing, respectively). Simulated altitude decreased RR interval (from 879 +/- 45 to 770 +/- 39, P < 0.01) and increased indices deducted from spectral analysis of heart rate variability (low frequency/high frequency (LF/HF) ratio from 1.6 +/- 0.5 to 3.2 +/- 1.1, P < 0.05) and systolic blood pressure (low frequency fluctuations from 2.30 +/- 0.31 to 3.07 +/- 0.24 In-mmHg2, P< 0.05) in controls, indicating sympathetic activation; these changes were blunted in yoga trainees, and in both groups during slow or yogic breathing. No effect of altitude was seen on stimulation of carotid baroreceptors in both groups. CONCLUSIONS: Well-performed slow yogic breathing maintains better blood oxygenation without increasing VE (i.e. seems to be a more efficient breathing) and reduces sympathetic activation during altitude-induced hypoxia. PMID- 11393680 TI - Angiotensin converting enzyme is the main contributor to angiotensin I-II conversion in the interstitium of the isolated perfused rat heart. AB - OBJECTIVES: Recent studies in homogenized hearts suggest that chymase rather than angiotensin converting enzyme (ACE) is responsible for cardiac angiotensin I to angiotensin II conversion. We investigated in intact rat hearts whether (i) enzymes other than ACE contribute to angiotensin I to angiotensin II conversion and (ii) the localization (endothelial/extra-endothelial) of converting enzymes. DESIGN AND METHODS: We used a modified version of the rat Langendorff heart, allowing separate collection of coronary effluent and interstitial fluid. Hearts were perfused with angiotensin I (arterial concentration 5-10 pmol/ml) under control conditions, in the presence of captopril (1 micromol/l) or after endothelium removal with 0.2% triton X-100. Endothelium removal was verified as the absence of a coronary vasodilator response to 10 nmol bradykinin. Angiotensin I and angiotensin II were measured in coronary effluent and interstitial fluid with sensitive radioimmunoassays. RESULTS: In control hearts, 45% of arterial angiotensin I was metabolized during coronary passage, partly through conversion to angiotensin II. At steady-state, the angiotensin I concentration in interstitial fluid was three to four-fold lower than in coronary effluent, while the angiotensin II concentrations in both fluids were similar. Captopril and endothelium removal did not affect coronary angiotensin I extraction, but increased the interstitial fluid levels of angiotensin I two- and three-fold, respectively, thereby demonstrating that metabolism (by ACE) as well as the physical presence of the endothelium normally prevent arterial angiotensin I from reaching similar levels in coronary effluent and interstitial fluid. Captopril, but not endothelium removal, greatly reduced the angiotensin II levels in coronary effluent and interstitial fluid. With the ACE inhibitor, the angiotensin II/I ratios in coronary effluent and interstitial fluid were 83 and 93% lower, while after endothelium removal, the ratios were 33 and 71% lower. CONCLUSIONS: In the intact rat heart, ACE is the main contributor to angiotensin I to angiotensin II conversion, both in the coronary vascular bed and the interstitium. Cardiac ACE is not limited to the coronary vascular endothelium. PMID- 11393681 TI - Left ventricular function in mice lacking the AT2 receptor. AB - OBJECTIVES: The role of the AT2 receptor in the heart is incompletely understood. We investigated left ventricular performance in AT2 receptor knockout mice, with and without deoxycorticosterone acetate (DOCA)-salt treatment. Given the putative opposing functions of the AT1 and AT2 receptor, we also analysed AT1 receptor expression in the left ventricle. METHODS: We used a miniaturized conductance manometer system to measure pressure-volume loops for analysing left ventricular performance under baseline conditions and after increasing peripheral vascular resistance. We determined left ventricular AT1-receptor expression by RNase protection assays. RESULTS: In AT2 receptor knockout mice, end-systolic and end diastolic volumes were lower than in wild-type mice, so that pressure-volume loops were shifted leftward. Left ventricular systolic and diastolic kinetics were not different between the groups. AT2 receptor knockout mice and wild-type mice both stabilized their reduced stroke volume after laparatomy as peripheral resistance was increased. DOCA-salt treatment increased elastance in AT2 receptor knockout mice, compared to controls. Furthermore, AT2 receptor knockout mice had a steeper increase in dP/dtmax. Left ventricular AT1 receptor gene expression was increased in AT2 receptor knockout mice and was not down-regulated in response to DOCA-salt treatment. Finally, the hearts of AT2 receptor knockout mice were smaller than controls, but increased in size in response to DOCA-salt treatment. CONCLUSIONS: AT2 receptor knockout mice displayed no major changes in left ventricular function at baseline or in response to DOCA-salt treatment, compared to wild-type mice. The AT2 receptor may be important to AT1 receptor expression in response to DOCA-salt challenge and may have some influence on cardiac growth responses. PMID- 11393682 TI - Heart rate-lowering calcium antagonists in hypertensive post-myocardial infarction patients. AB - OBJECTIVES: To analyse effects of a heart rate-lowering calcium antagonist in hypertensive post-myocardial infarction patients. DESIGN AND METHODS: From three large, randomized, placebo-controlled, secondary prevention trials investigating verapamil or diltiazem (the first and second Danish Verapamil Infarction Trials and the Multicentre Diltiazem Post-Infarction Trial) data from a total of 1,325 hypertensive post-myocardial infarction patients (drugs = 667, placebo = 658) were pooled to assess effect of blinded therapy on mortality and event rates. RESULTS: Treatment with heart rate-lowering calcium antagonists was associated with significant reduction in event rates [21.4 versus 27.4%; risk ratio (RR) = 0.76, confidence interval (CI) = 0.61 -0.95, P= 0.013]. Mortality rates in the treatment group were 15.1 versus 17.5% in the control group (RR = 0.87, CI = 0.66 1.13, P= 0.296). Among the subset of 964 hypertensive patients without pulmonary congestion, there was some reduction in mortality rate (11.3 versus 15.3% in the control group; RR = 0.72, P= 0.066) and significant reduction in event rates (18 versus 24.4% for control group; RR = 0.70, P= 0.011). In patients with pulmonary congestion and hypertension, however, calcium antagonists were associated with a 25% increase in mortality (RR = 1.25, P= 0.339), while event rate RR was 1.00. After an adjustment for significant covariates, RR for mortality in treatment versus control groups was 0.76 (P= 0.159). For event rates, RR was 0.74 (P= 0.057). CONCLUSIONS: Heart rate-lowering calcium antagonists decrease event rates in hypertensive post-myocardial infarction patients, but only in those without pulmonary congestion. PMID- 11393683 TI - Enhanced renal microvascular reactivity to angiotensin II in hypertension is ameliorated by the sulfonimide analog of 11,12-epoxyeicosatrienoic acid. AB - OBJECTIVES: Epoxygenase metabolites produced by the kidney affect renal blood flow and tubular transport function and 11,12-epoxyeicosatrienoic acid (11,12 EET) has been putatively identified as an endothelium-derived hyperpolarizing factor. The current studies were performed to determine the influence of 11,12 EET on the regulation of afferent arteriolar diameter in angiotensin II-infused hypertensive rats. MATERIALS AND METHODS: Male Sprague-Dawley rats received angiotensin II (60 ng/min) or vehicle via an osmotic minipump. Angiotensin II infused hypertensive and vehicle-infused normotensive rats were studied for 2 weeks following implantation of the minipump. Renal microvascular responses to the sulfonimide analog of 11,12-EET (11,12-EET-SI) and angiotensin II were observed utilizing the in-vitro juxtamedullary nephron preparation. Renal cortical epoxygenase enzyme protein levels were quantified by Western blot analysis. Renal microvessels were also isolated and epoxygenase metabolite levels measured by negative ion chemical ionization (NICI)/gas chromatography-mass spectroscopy. RESULTS: Systolic blood pressure averaged 118 +/- 2 mmHg prior to pump implantation and increased to 185 +/- 7 mmHg in rats infused with angiotensin II for 2 weeks. Afferent arteriolar diameters of 2-week normotensive animals averaged 22 +/- 1 microm. Diameters of the afferent arterioles were 17% smaller in hypertensive rats (P< 0.05); however, arterioles from both groups responded to 11,12-EET-SI (100 nmol) with similar 15-17% increases in diameter. As we previously demonstrated, the afferent arteriolar reactivity to angiotensin II was enhanced in angiotensin II-infused animals. Interestingly, elevation of 11,12-EET-SI levels to 100 nmol reversed the enhanced vascular reactivity to angiotensin II associated with angiotensin II hypertension. Renal microvascular EET levels were not different between groups and averaged 81 +/- 9 and 87 +/- 13 pg/mg per 30 min in normotensive and hypertensive animals, respectively. Renal cortical microsomal levels of the epoxygenase CYP2C23 and CYP2C11 proteins were also similar in normotensive and angiotensin II hypertensive rats. CONCLUSIONS: Taken together, these data support the concept that renal microvascular 11,12-EET activity and levels may not properly offset the enhanced angiotensin II renal vasoconstriction during angiotensin II hypertension. PMID- 11393685 TI - Genetic identification of Saccharomyces bayanus var. uvarum, a cider-fermenting yeast. AB - Twenty-one Saccharomyces strains isolated from a cider process were analysed in terms of karyotypes, Y' S. cerevisiae sequence occurrence, rDNA structure and cross-fertility with species tester strains. A strong predominance of S. bayanus var. uvarum G. Naumov was found (18 strains vs. three S. cerevisiae). Among the S. bayanus var. uvarum, only three strains proved to contain species-specific Y' S. cerevisiae sequences. PMID- 11393684 TI - AT oligonucleotides inducing B lymphocyte activation exist in probiotic Lactobacillus gasseri. AB - This study determined oligonucleotide sequences of mitogenic DNA derived from lactic acid bacteria (LAB). The chromosomal DNA, which was purified from 12 out of 16 strains of Lactobacillus acidophilus group LAB, induced proliferation of splenic lymphocytes. When DNA from L. gasseri JCM1131T was cloned and amplified using PCR, the mitogenic activities of B lymphocytes were significantly increased by 108 of 321 DNA clones. Ten high homologous nucleotide sequences were found as possible DNA sequences of mitogens, and were then chemically synthesized (sOL-LG1 to sOL-LG10). Two nucleotide sequences (sOL-LG7 and sOL-LG10) that consist of only A and T nucleotides (AT oligonucleotides) were characterized as B lymphocyte specific mitogens because they resulted in proliferation of B lymphocytes but not of T lymphocytes. sOL-LG7 preferentially bound to large B lymphocytes and enhanced the expression of the CD86 antigen more than the CD69 antigen on B lymphocytes. The findings show that mitogenic AT oligonucleotides are likely to restrict pre-activated subsets of B lymphocytes. This study demonstrated that novel AT oligonucleotides triggering B lymphocyte mitogenic responses exist in the nucleoids of L. gasseri and proposed that they have potential as applicants for the production of new functional foods, "Bio-Defense Foods". PMID- 11393686 TI - Effect of freeze drying and protectants on viability of the biocontrol yeast Candida sake. AB - The effects of freezing method, freeze drying process, and the use of protective agents on the viability of the biocontrol yeast Candida sake were studied. Freezing at -20 degrees C was the best method to preserve the viability of C. sake cells after freeze drying using 10% skim milk as a protectant (28.9% survival). Liquid nitrogen freezing caused the highest level of damage to the cells with viability < 10%. Different concentrations of exogenous substances including sugars, polyols, polymers and nitrogen compounds were tested either alone or in combination with skim milk. There was little or no effect when additives were used at 1% concentration. Galactose, raffinose and sodium glutamate at 10% were the best protective agents tested alone but the viability of freeze-dried C. sake cells was always < 20%. Survival of yeast cells was increased from 0.2% to 30-40% by using appropriate protective media containing combinations of skim milk and other protectants such as 5% or 10% lactose or glucose, and 10% fructose or sucrose. PMID- 11393687 TI - Characterization of Penicillium roqueforti strains used as cheese starter cultures by RAPD typing. AB - Seventy-six strains of Penicillium roqueforti used as starter cultures for mould ripened blue cheeses have been analysed for their RAPD genotype by using three different primers. A comparison of the RAPD patterns within each primer group revealed that the genetic constitution of the strains was similar, as most of the strains showed very similar overall patterns. Despite these similarities with each primer, distinct RAPD genotype groups could be identified. With one of the primers, it was possible to detect two heteropolymorphic DNA regions resulting in 13 different groups. With the other two primers, three or four groups could be identified. Between the groups of the different primers marked correspondence with respect to strain distribution could be observed, indicating that the polymorphisms detected by the primers were not independent. The RAPD patterns were compared to the production of secondary metabolites. A correlation was observed between the RAPD patterns of all primers and the production of mycophenolic acid. In addition, one of the primer (ari1) was able to distinguish between P. roqueforti strains producing larger or smaller numbers of metabolites. PMID- 11393688 TI - Escherichia coli O157:H7 in faeces from cattle, sheep and pigs in the southwest part of Norway during 1998 and 1999. AB - During a 2-year period from January 1998 to December 1999, intestinal content from 1541 cattle, 665 sheep and 1976 pigs were analysed for Escherichia coli O157:H7 using the immunomagnetic separation procedure. The animals originated from 848, 605 and 832 herds from the southwest part of Norway, respectively. E. coli O157:H7 was present in three samples from cattle from different herds, giving a herd prevalence of 0.35% and an animal prevalence of 0.19%. From pigs, E. coli O157:H7 was isolated from two pigs from different herds, giving a herd prevalence of 0.24% and an animal prevalence of 0.1%. A follow-up study revealed another positive testing pig from one of these herds. E. coli O157:H7 was not found from any of the 665 investigated sheep. By PCR analysis, all six E. coli O157:H7 isolates were shown to contain the genes encoding Shiga toxin 2 (stx2), the intimin protein (eae) and the H7 flagellum (fliC-H7). One of the cattle isolates also harboured the Shiga toxin 1 encoding (stx1) gene. The six isolates were differentiated into three pulse-field gel electrophoresis profiles. The results indicate that the occurrence of E. coli O157:H7 in cattle, sheep and pigs in the southwest part of Norway is low compared to other European countries. PMID- 11393689 TI - Can food-related environmental factors induce different behaviour in two key serovars, 4b and 1/2a, of Listeria monocytogenes? AB - Listeria monocytogenes isolates (81 in total; 42 isolated from cases of human listeriosis: 39 isolated from food), belonging to serovars 1/2a or 4b, were studied for any group differences between serovars to selected factors associated with foods (two bacteriocins and mild heat treatment), growth kinetics at 37 degrees C and pathogenicity for chick embryos. The isolates were tested for sensitivity to two bacteriocins at 4 degrees C and 37 degrees C, and were tested for the remaining parameters both before and after exposure to cold storage (4 degrees C) with starvation. In addition, the isolates were typed using pulsed field gel electrophoresis (PFGE), multilocus enzyme electrophoresis (MEE) and phage typing to find any correlation between the types and group differences in the chosen parameters. Considerable strain diversity within each L. monocytogenes serovar with respect to the chosen parameters was observed, especially after exposure to cold storage. Nevertheless, the serovar 1/2a isolates, as a group, tended to be more resistant to the two antilisterial bacteriocins at 4 degrees C than the group of serovar 4b isolates. In contrast, after cold storage at 4 degrees C, L. monocytogenes serovar 4b isolates, as a group, tended to be more resistant to heat treatment at 60 degrees C than the group of 1/2a isolates. In addition, the serovar 4b group tended to have shorter lag phases and higher pathogenicity, when transferred from cold storage to body temperature (37 degrees C), than the group of serovar 1/2a isolates. No correlation between PFGE-, MEE- and phage-types and the tested parameters was found. Although the above serovar related differences were observed only when mean values of the groups were compared (not all isolates within each group followed the group pattern), the results indicate interesting directions for further research. PMID- 11393690 TI - Characteristics and significance of yeasts' involvement in cassava fermentation for 'fufu' production. AB - Six different strains of yeast, namely Candida krusei, C. tropicalis, Pichia saitoi, Saccharomyces cerevisiae, P. anomala and Zygosaccharomyces bailii were found present in cassava-fermenting water in the early part of the fermentation. The latter part of the fermentation was dominated in all cases by three strains of yeast namely C. krusei, C. tropicalis and Z. bailii. All the yeast strains exhibited amylolytic capabilities while none was able to produce cellulase. All the strains except Zygosaccharomyces spp. exhibited polygalacturonase activity, but only C. krusei was able to produce linamarase. In a study on the inter relationships between C. krusei and Lactobacillus plantarum, the growth of the yeast strain was not enhanced in cassava by the presence of the lactic acid bacteria, but the growth of the L. plantarum strain was significantly enhanced when co-inoculated with C. krusei. PMID- 11393691 TI - Bibliography of food microbiology. PMID- 11393692 TI - Determination of minocycline in human plasma by high-performance liquid chromatography coupled to tandem mass spectrometry: application to bioequivalence study. AB - Minocycline was determined in human plasma by HPLC-MS-MS using clarithromycin as an internal standard. The method is fast (single liquid extraction and run time of <3 min) and sensitive (5 ng/ml) and it was employed in a bioequivalence study of two 100 mg tablet formulations in 24 healthy volunteers. The 90% confidence interval of the individual ratio geometric mean for both AUC(0-96 h) and Cmax were 99.2-111.1% and 95.6-117.5%, respectively. Thus, Minoderm was considered bioequivalent to Minomax according to both the rate and extent of absorption. No food interaction was observed with either formulation. PMID- 11393693 TI - High-performance liquid chromatographic determination of bradykinin in saliva: a critical review and a new method. AB - Because of difficulties or dubious results with previously published methodologies, a new semi-automated HPLC method with UV absorbance detection was developed and applied to the determination of bradykinin (BK) in human saliva. The new method consisted of an uncomplicated sample preparation involving the addition to saliva of an equal volume of 0.1 M orthophosphoric acid to stabilize BK, vortex-mixing, centrifugation, and separation, followed by chromatography of the supernatant phase on a C8, 150x3.9-mm (I.D.) stainless steel column. The mobile phase was composed of 19% acetonitrile/0.1% trifluoroacetic acid at flow rate of 0.4 ml/min. Using UV detection at 220 nm, the detection limit was 1 ng/ml for the BK standard, and 7 ng/ml for the assay of endogenous salivary BK. The orthophosphoric acid initially added to the saliva allowed BK to be stabilized from enzymic degradation at 20 degrees C for 5 days and at 4 degrees C for 60 days. Assignment made to the peak with the chromatographic properties of salivary BK was confirmed by HPLC-MS with an electrospray interface. This paper presents a new method that is reproducible, reliable and allows kinetic studies of salivary BK to be performed for clinical investigations. PMID- 11393695 TI - Lamotrigine analysis in blood and brain by high-performance liquid chromatography. AB - A reversed-phase high-performance liquid chromatography assay was developed and validated to determine plasma and brain lamotrigine concentrations allowing pharmacokinetic-pharmacodynamic studies of this new antiepileptic drug in patients and laboratory animals. Lamotrigine and its internal standard were extracted, under alkaline conditions, from plasma and brain homogenate, into ethyl acetate; brain proteins were previously precipitated with trichloroacetic acid. The method was linear between 0.1 and 15.0 mg/l for plasma, with a detection limit of 0.008 mg/l, and between 0.1 and 5.0 mg/l for brain homogenate, with a detection limit of 0.023 mg/l. The method proved to be simple, useful and appropriate, not only for clinical and experimental research, but also for routine monitoring of lamotrigine concentrations in patients. PMID- 11393694 TI - Separation of carvedilol enantiomers in very small volumes of human plasma by capillary electrophoresis with laser-induced fluorescence. AB - A sensitive capillary electrophoretic method for the determination of carvedilol enantiomers in 100 microl of human plasma has been developed and validated. Carvedilol and the internal standard carazolol are isolated from plasma samples by liquid-liquid extraction using diethylether. A sensitive and selective detection is provided by helium-cadmium laser-induced fluorescence. The total analysis time is 17.5 min, about 30 min are needed for the sample preparation. The linearity of the assay ranges from 1.56 to 50 ng/ml per carvedilol enantiomer. The limits of quantification (LOQ) for the carvedilol enantiomers in 100 microl of human plasma are 1.56 ng/ml. The inter-day accuracy for R carvedilol is between 95.8 and 103% (104% at LOQ) and for S-carvedilol between 97.1 and 103% (107% at LOQ); the inter-day precision values are between 3.81 and 8.64% (10.9% at LOQ) and between 5.47 and 7.86% (7.91% at LOQ) for R- and S carvedilol, respectively. The small sample volume needed is especially advantageous for the application in clinical studies in pediatric patients. As an application of the assay concentration/time profiles of the carvedilol enantiomers in a 5-year-old patient receiving a test dose of 0.09 mg/kg carvedilol are reported. PMID- 11393696 TI - Rapid and sensitive high-performance liquid chromatographic method for simultaneous determination of retinol, alpha-tocopherol, 25-hydroxyvitamin D3 and 25-hydroxyvitamin D2 in human plasma with photodiode-array ultraviolet detection. AB - A new rapid and sensitive high-performance liquid chromatographic method using 0.5 ml of plasma has been developed for the simultaneous determination of retinol (vitamin A), alpha-tocopherol (vitamin E), 25-hydroxyvitamin D2 and 25 hydroxyvitamin D3. The eluate was monitored with a photodiode-array detector with two fixed wavelengths (267 nm for vitamin D, 292 nm for alpha-tocopherol and retinol). For all compounds, including internal standards, the method provides extraction recoveries greater than 81%. Detection limits were equal to or lower than 1.5 microg/l for the 4 vitamins. Linearity of standards was excellent (r>0.999 in all cases). Intra-day and inter-day precision were generally acceptable; the intra-dayassay C.V. was 3/4 7.7 for all compounds and the inter day-assay C.V. was <9.2% except for the lower concentrations of 25-hydroxyvitamin D3, 25-hydroxyvitamin D2 and alpha-tocopherol (10.8, 11.8 and 11.9, respectively). The important properties of the present method are its ease of use, its rapidity, since sample preparation was achieved in 15 min and all the compounds were eluted in less than 15 min, and its small sample volume required (=0.5 ml), which enables it to be used in pediatric practice. PMID- 11393697 TI - Stir bar sorptive extraction-thermal desorption-capillary gas chromatography-mass spectrometry applied to the analysis of polychlorinated biphenyls in human sperm. AB - Stir bar sorptive extraction (SBSE) on polydimethylsiloxane (PDMS) was applied to the enrichment of polychlorinated biphenyls (PCBs) from human sperm. The seven Ballschmiter PCBs were used as model compounds. The extracted PCBs were then thermally desorbed from the stir bar and analysed on-line by capillary gas chromatography (CGC) with mass spectrometric detection (MS). Method development started with the analysis of PCBs spiked in water. Methanol had to be added to the samples in order to reduce the influence of glass adsorption on recovery and reproducibility. Recoveries in water for all PCBs varied around 50-60% and were limited for low molecular mass (MM) PCBs by polarity changes in the sample due to methanol addition and for high MM PCBs by non-equilibrium conditions. Matrix suppression by the lipophilic medium lowered the recoveries in the sperm samples proportional with PCB polarity. The method was validated and although limits of detection (LOD) for the individual congeners were in the sub-ppt level ( or = 0.996) with accuracy < or =9.8% and precision < or = 13.2%. Total imprecision at quantification limits was 15.2% at 10 ng/ml and 16.3% at 1,500 ng/ml for MMF and 15.1% at 21.0 ng/ml and 17.5% at 1,300 ng/ml for MPA. This HPLC-MS method will be applicable to the profiling of MMF amounts in skin and its conversion to MPA after application of different formulations. PMID- 11393726 TI - Multi-residue analysis of avermectins in swine liver by immunoaffinity extraction and liquid chromatography-mass spectrometry. AB - A multi-residue LC-MS method was developed to determine avermectin residues in swine liver. Abamectin and ivermectin (22,23-dihydroabamectin) were extracted, and cleaned up by immunoaffinty columns with immobilized anti-avermectin polyclonal antibodies. The cleaned samples were separated by high-performance liquid chromatography (HPLC) with a C8 column and determined by negative-ion atmospheric pressure chemical ionization (APCI) mass spectrometry (MS) using selective ion monitoring (SIM) of [M-H]-. Recoveries of abamectin and ivermectin from fortified samples at 5-100 microg kg(-1) levels ranged from 74 to 94% and from 65 to 87%, respectively. The limits of detection were 5 microg of abamectin or ivermectin in 1 kg sample. PMID- 11393727 TI - Purification of factor X by hydrophobic interaction chromatography. AB - Human factor X has been purified to homogeneity by hydrophobic interaction chromatography on phenyl-sepharose. The coagulation protein did not interact with the resin in the presence of 2-3 M NaCl whereas contaminants were retained. This single purification step, in conjunction with classical purification strategies, is a powerful tool in generating high purity factor X and is based on resins which are readily available. PMID- 11393728 TI - A dual-mode approach to the selective separation of antibodies and their fragments. AB - A novel chromatography method for the separation of antibodies is described. The adsorption of antibodies on the solid phase involves interaction with a ligand that combines mild hydrophobic characteristics and some degree of molecular recognition with a derivative of pyridine. This combined effect results in the adsorption of antibodies in the absence of lyotropic salts. When environmental pH is changed, the ligand becomes ionically charged, allowing the desorption of antibodies. The mechanism of adsorption, involving hydrophobic associations and ionic related interaction, is here qualified as dual-mode. Studies on the determination of the apparent dissociation constant for immunoglobulins G are presented. Adsorption of antibodies from crude feedstocks typically occurs without adjustment of pH or ionic strength. The sorbent is then washed with a buffer to eliminate protein impurities and, when lowering the environmental pH, antibodies are desorbed. The solid-phase material is used for the separation of antibodies from an ascites fluid and from a cell culture supernatant, followed by a polishing step on an hydroxyapatite column. Preliminary studies, related to the ability of the solid phase to separate antibody fragments, are also reported. In these studies, it has been demonstrated that both Fab and Fc fragments from polyclonal IgG are adsorbed to the solid phase under typical binding conditions. Under other defined physico-chemical conditions (ionic strength and pH), separation of both fragments in a single step has been achieved. PMID- 11393729 TI - A new and rapid method for monitoring the new oxazolidinone antibiotic linezolid in serum and urine by high performance liquid chromatography-integrated sample preparation. AB - A sensitive and rapid HPLC-assay for determining the new oxazolidinone antibiotic linezolid in serum and urine is described. HPLC-integrated sample preparation permits the direct injection of serum and urine samples without any pre treatment. The in-line extraction technique is realized by switching automatically from the extraction column to the analytical column. After the matrix has passed the extraction column the retained analyte will be quantitatively transferred to the analytical column where separation by isocratic HPLC will be performed. Linezolid is detected according to its absorption maximum at 260 nm. The quantification limits are estimated to be 0.3 and 0.5 microg/ml in serum and urine samples, respectively. The described procedure allows sample clean-up and determination of the antibiotic within 20 min, thereby facilitating drug-monitoring in clinical routine. PMID- 11393730 TI - Extraction and quantification of epibatidine in plasma. AB - Epibatidine was extracted from human and mouse plasma into a hexane-isopropanol mixture and back-extracted into a phosphate buffer, pH 2.5, then identified by HPLC isocratically using a CN column and quantified with ultraviolet detection at a fixed wavelength of 214 nm. The percent recovery of epibatidine from spiked plasma samples was 83.6% and the percent extraction was linear between 10 and 1,000 ng/ml. Desipramine was used as the internal standard. For spiked control samples containing 50 and 750 ng/ml, between-day precisions were 20.8 and 7.2% (RSD%), respectively; accuracy was 87.0 and 99.1%, respectively. The limit of detection was 2 ng/ml. Using this method, an intraperitoneal dose of 0.1 mg/kg of epibatidine produced mean levels of 7.3 and 37.1 ng/ml in pooled male and female plasma samples from C57BL/10 J mice, respectively. This is a simple and straightforward procedure by which plasma samples may be analyzed for epibatidine. PMID- 11393731 TI - Determination of ketorolac in human plasma by reversed-phase high-performance liquid chromatography using solid-phase extraction and ultraviolet detection. AB - An improved high-performance liquid chromatographic method has been developed to measure human plasma concentrations of the analgesic nonsteroidal anti inflammatory drug ketorolac for use in pharmacokinetic studies. Samples were prepared for analysis by solid-phase extraction using Bond-Elut PH columns, with nearly complete recovery of both ketorolac and the internal standard tolmetin. The two compounds were separated on a Radial-Pak C18 column using a mobile phase consisting of water-acetonitrile-1.0 mol/l dibutylamine phosphate (pH 2.5) (30:20:1) and detected at a UV wavelength of 313 nm. Using only 250 microl of plasma, the standard curve was linear from 0.05 to 10.0 microg/ml. PMID- 11393732 TI - Liquid chromatography-tandem mass spectrometry identification of metabolites of two 5-HT1A antagonists, N-[2-[4-(2-methoxylphenyl)piperazino]ethyl]-N-(2-pyridyl) trans- and cis-4-fluorocyclohexanecarboxamide, produced by human and rat hepatocytes. AB - Two 5-HT1A antagonists, t-FCWAY and c-FCWAY, were developed as imaging agents for positron emission tomography (PET). In order to evaluate these compounds, hepatocytes from both human and rat were utilized to produce metabolites and LC MS-MS was used to identify metabolites. These in vitro metabolism studies indicate that hydrolysis of the amide linkage is the major metabolism pathway for humans, whereas aromatic ring-oxidation is the major metabolism pathway for rat. The rat hepatocyte results correlate well with in vivo rat metabolism studies. Based on the structures of the metabolites, we have developed an extraction procedure to determine the concentration of the parent compound in plasma. PMID- 11393733 TI - Study on the electrochemical detection of the macrolide antibiotics clarithromycin and roxithromycin in reversed-phase high-performance liquid chromatography. AB - The optimal conditions of the amperometric detection of the macrolide antibiotics clarithromycin and roxithromycin were found by cyclic voltammetric studies and HPLC-electrochemical detection responses obtained in different temperatures (25.5 60 degrees C) and different but almost isoelutropic binary, ternary and quaternary mixtures of aqueous buffer (pH 7), methanol, acetonitrile and isopropanol. These conditions were also proved to be applicable for the quantitative detection of clarithromycin in human plasma using roxithromycin as an internal standard and vice versa. It was demonstrated that increased attention has to be paid to eluent composition and column temperature to ensure sensitive and reproducible electrochemical responses as well as regularly shaped peaks for both macrolides tested. PMID- 11393734 TI - Quantitation of mitomycin C in human ocular tissues by high-performance liquid chromatography-photo-diode array detection. AB - A chromatographic method, which can quantitate mitomycin C (MMC) along with two antiglaucoma drugs, is described. The separation of MMC, alphagan and timolol was performed on a reversed-phase C18 column with water-methanol-trifluoroacetic acid (65:35:0.01, v/v) as the mobile phase. By monitoring at 360, 248 and 296 nm, the lower limits of detection for MMC, alphagan and timolol are, respectively, 1.0, 2.0 and 5.0 ng (injection amount) at three-time S/N ratio. The dynamic ranges of quantitation for the three drugs are, respectively, 1.0 ng-10.0 microg, 2.0 ng 10.0 microg and 5.0 ng-10.0 microg with linearity being larger than 0.9960. This method was applied to the determination of MMC levels in Tenon's and trabeculum tissues of 10 glaucoma patients. MMC levels in these tissues, which were obtained from glaucoma filtering surgery, were determined following a multiple extraction with methanol. The recovery of MMC for a two-batch extraction was better than 91.2%. The reproducibility of measurement for the MMC levels in these tissues is 2.5-6.0% RSD for triplicate injections. The intra-day variation of retention times for the MMC peaks was less than 1.6% RSD (n=3). The inter-day variation of retention times for the MMC peaks was less than 4.8% RSD (n=3). MMC was detectable in three trabeculum tissues out of 10 cases (ranging from 0.8 to 25.5 ng/mg specimen), while MMC was detected in nine Tenon's tissues out of 10 cases (ranging from 0.3 to 21.1 ng/mg specimen). The results obtained show that the method is sensitive and selective for the quantitation of MMC. PMID- 11393735 TI - Extractionless method for the simultaneous high-performance liquid chromatographic determination of urinary caffeine metabolites for N acetyltransferase 2, cytochrome P450 1A2 and xanthine oxidase activity assessment. AB - Urinary metabolic ratios of caffeine are used in humans to assess the enzymatic activities of cytochrome P450 isoenzyme 1A2 (CYP1A2), xanthine oxidase (XO) and for phenotyping individuals for the bimodal N-acetyltransferase 2 (NAT2), all of them involved in the activation or detoxification of various xenobiotic compounds. Most reported analytical procedures for the measurement of the urinary metabolites of caffeine include a liquid-liquid extraction of urine samples prior to their analysis by reversed-phase HPLC. At neutral to basic pH however, 5 acetylamino-6-formylamino-3-methyluracil (AFMU), a metabolite of caffeine, spontaneously decomposes to 5-acetylamino-6-amino-3-methyluracil (AAMU). Since AAMU is not extracted in most organic solvents, the extent of AFMU decomposition cannot be precisely assessed. Although the decomposition reaction can be minimized by immediate acidification of the urine, accurate results can only be obtained when both AAMU and AFMU are monitored, or alternatively, if AAMU is measured after complete transformation of AFMU into AAMU in basic conditions. We report a liquid chromatographic method for the simultaneous quantitative analysis of the five urinary metabolites of caffeine used for the CYP1A2, XO and NAT2 phenotyping studies: AAMU, AFMU, 1-methylxanthine, 1-methyluric acid and 1,7 dimethyluric acid. These metabolites are satisfactory separated from all other known caffeine metabolites as well as endogenous urinary constituents. Sample treatment does not require any liquid-liquid extraction procedure. Urine samples are diluted and centrifuged before being injected (10 microl) onto a YMC-Pack Polyamine II (250x4.6 mm) column. A step-wise gradient elution program is applied using acetonitrile-0.75% (v/v) formic acid: (91:9) at 0 min-->(75:25) at 25 min- >(65:35) at 35 min-->(65:35) at 45 min, followed by a re-equilibration step to the initial solvent composition. The flow-rate is 1.0 ml/min and the separations are monitored by UV absorbance at 260 and 280 nm. The procedure described here represents a substantial improvement over previous methods: a single analysis and a minimal urine sample treatment enables the simultaneous quantitation of five caffeine metabolites, notably AFMU and AAMU, used for the determination of CYP450 1A2, XO and NAT2 enzyme activity. Importantly enough, phenotyping individuals for the bimodal NAT2 is made possible without the uncertainty associated with the deformylation of AFMU, which is likely to happen at all steps prior to the analysis, during sample storage and even in the bladder of the subjects. PMID- 11393736 TI - Simultaneous determination of the HIV protease inhibitors indinavir, amprenavir, saquinavir, ritonavir and nelfinavir in human plasma by high-performance liquid chromatography. AB - A reversed-phase high-performance liquid chromatographic method for the simultaneous quantitative determination of five HIV protease inhibitors (i.e. indinavir, amprenavir, saquinavir, ritonavir and nelfinavir) in human plasma is described. An aliquot of 500 microl plasma was extracted with 0.5 ml of 0.1 M NH4OH and 5 ml of methyl tert.-butyl ether. After evaporating, the residue was dissolved in eluent mixture of acetonitrile and 50 mM KH2PO4 adjusted to pH 5.6 with 50 mM Na2HPO4 (43:57, v/v). Subsequently, the eluent was washed with hexane. Chromatography was performed using a C18 reversed-phase column. Ultraviolet detection at 215 nm was used. Linearity of the method was obtained in the concentration range of 0.05-20 microg ml(-1) for all five protease inhibitors. Our method is now in use to analyse plasma samples from patients treated with co administration of HIV protease inhibitors. PMID- 11393737 TI - Determination of bisphenol A in blood using high-performance liquid chromatography-electrochemical detection with solid-phase extraction. AB - A method for the determination of bisphenol A (BPA) in blood was investigated using high-performance liquid chromatography-electrochemical detection (HPLC-ED) with solid-phase extraction. When BPA at the concentrations of 25-100 ng/ml were added to whole blood, BPA recoveries were 26-48%. When BPA was added to water, plasma or hemolyzed red blood cells (H-RBC), BPA recoveries in water and plasma were almost similar (94%). However, the recovery in H-RBC was very low (36-46%). When BPA and plasma were added to H-RBC, the recovery was 70-85%. In authentic bovine metHb solution, BPA decreased depending on the metHb concentration, however, BPA recovery in the solution added with more than 17% plasma was higher than that in metHb only. These suggest that metHb influences the BPA recovery in whole blood. However, an accurate determination of BPA using HPLC was easily made possible by separating RBC from plasma. PMID- 11393738 TI - Use of non-covalent labeling in illustrating ligand binding to human serum albumin via affinity capillary electrophoresis with near-infrared laser induced fluorescence detection. AB - This paper demonstrates the use of a near-infrared (NIR) dye as a non-covalent label for human serum albumin (HSA). The dye is a water soluble, heptamethine cyanine dye. The utility of the dye as a tracer illustrating the binding of various drugs to HSA is demonstrated via affinity capillary electrophoresis with near-infrared laser-induced fluorescence detection (ACE-NIR-LIF). Additionally, the factors affecting the separation of relevant species were investigated. The change in quantum yield of the dye upon complexation with HSA was calculated. Spectrophotometric measurements were conducted to study the stoichiometry of the dye albumin complex. PMID- 11393739 TI - Alternative methods to examine hospital efficiency: data envelopment analysis and stochastic frontier analysis. AB - There has been increasing interest in the ability of different methods to rank efficient hospitals over their inefficient counterparts. The UK Department of Health has used three cost indices to benchmark NHS hospitals (Trusts). This study uses the same dataset and compares the efficiency rankings from the cost indices with those obtained using Data Envelopment Analysis (DEA) and Stochastic Frontier Analysis (SFA). The paper concludes that the methods each have particular strengths and weaknesses and potentially measure different aspects of efficiency. Several specifications should be used to develop ranges of inefficiency to act as signalling devices rather than point estimates. It is argued that differences in efficiency scores across different methods may be due to random "noise" and reflect data deficiencies. The conclusions concur with previous findings that there are not truly large efficiency differences between Trusts and savings from bringing up poorer performers would in fact be quite modest. PMID- 11393740 TI - A simulation modelling approach to evaluating alternative policies for the management of the waiting list for liver transplantation. AB - A shortage of donor liver grafts unfortunately results in approximately 10% of patients dying whilst listed for a liver transplant in Europe and the United States. Thus it is imperative that all available organs are used as efficiently as possible. This paper reports upon the application of a simulation modelling approach to assess the impact of several alternative allocation policies upon the cost effectiveness of this technology at one liver transplant centre in the UK. The impact of changes in allocation criteria on the estimated net life expectancy, average net costs and overall cost effectiveness of the transplantation programme were evaluated. The incremental cost effectiveness ratio (ICER) for the base case allocation policy, based upon the time spent on the waiting list (i.e., longest wait first) was 11,557 pounds sterling at 1999 prices. The ICERs associated with an allocation policy based upon age (lowest age first), and an allocation policy based upon the severity of the pre-transplant condition of the patient (with most severely ill patients given a lower priority) were lower than the base case at 10,424 pounds sterling and 9,077 pounds sterling, respectively. The results of this modelling study suggest that the overall cost effectiveness of the liver transplantation programme could be improved if the current allocation policy were modified to give more weight to the age of the patient and the reduced chances of success of the most severely ill patients. PMID- 11393741 TI - Modeling and analysis of multistate access to elective surgery. AB - In this paper, we attempt to determine whether delays in scheduling operation affect waiting time in a queue for elective surgery. We analyze the waiting-list management system in a Canadian hospital. We estimate the impact of scheduling delays by modeling access to treatment as a multistate process. We found that patients with any delay in scheduling surgery had longer waiting times than patients without delays. For certain sources of delays, the admission rate was 50 60% lower compared with the rate for admissions without a delay independent of urgency of surgical intervention. Our findings support a concern that waiting time for elective surgery is not simply determined by how many patients are on the waiting list, or by how urgently they need treatment, but also by the waiting list management practice. PMID- 11393742 TI - Towards decision support for waiting lists: an operations management view. AB - This paper considers the phenomenon of waiting lists in a healthcare setting, which is characterised by limitations on the national expenditure, to explore the potentials of an operations management perspective. A reference framework for waiting list management is described, distinguishing different levels of planning in healthcare--national, regional, hospital and process--that each contributes to the existence of waiting lists through managerial decision making. In addition, different underlying mechanisms in demand and supply are distinguished, which together explain the development of waiting lists. It is our contention that within this framework a series of situation specific models should be designed to support communication and decision making. This is illustrated by the modelling of the demand for cataract treatment in a regional setting in the south-eastern part of the Netherlands. An input-output model was developed to support decisions regarding waiting lists. The model projects the demand for treatment at a regional level and makes it possible to evaluate waiting list impacts for different scenarios to meet this demand. PMID- 11393743 TI - Explaining inequality in the use of public health care services: evidence from Spain. AB - The aim of this article is to identify those factors which produce differences in the use of public health care services among individuals whose medical needs are similar. The empirical analysis employs Spanish data for three separate years and obtains results for four types of medical attention: visits to a GP, visits to a specialist, visits to the emergency services and hospital inpatient days. The results show that the influence of demand factors upon the probability of consuming more medical attention than expected depends on the type of health care considered.In addition, the minimal impact of supply factors is demonstrated. PMID- 11393744 TI - Cost inefficiency in Washington hospitals: a stochastic frontier approach using panel data. AB - We analyze a sample of Washington State hospitals with a stochastic frontier panel data model, specifying the cost function as a generalized Leontief function which, according to a Hausman test, performs better in this case than the translog form. A one-stage FGLS estimation procedure which directly models the inefficiency effects improves the efficiency of our estimates. We find that hospitals with higher casemix indices or more beds are less efficient while for profit hospitals and those with higher proportion of Medicare patient days are more efficient. Relative to the most efficient hospital, the average hospital is only about 67% efficient. PMID- 11393745 TI - Comparing teaching and non-teaching hospitals: a frontier approach (teaching vs. non-teaching hospitals). AB - This paper compares teaching and non-teaching hospitals in terms of their provision of patient services. We proceed by comparing the frontiers of the teaching and non-teaching hospitals using a data envelopment (DEA) type approach, which we apply to a sample of 236 teaching hospitals and 556 non-teaching hospitals operating in the US in 1994. Our results suggest that only about 10% of the teaching hospitals can effectively "compete" with non-teaching hospitals based on the provision of patient services. PMID- 11393746 TI - The technical efficiency of hospitals under a single payer system: the case of Ontario community hospitals. AB - Using DEA, we investigated the impact of ownership, size, and location on the relative technical efficiency of community hospitals in Ontario, which has a single payer system. Consistent with Hansmann's theory of non-profit organizations and contrary to US-based research, we find no significant differences in efficiency across ownership types (government, religious or secular non-profit). Nor do we find significant differences in efficiency by size or location. Our findings suggest that model formulation and differences in payer mix across types of hospitals in the US have a strong influence on the measurement of the hospital ownership-efficiency relationship. PMID- 11393747 TI - Effect of the raw extracts of Arthrinium strains (Hyphomycetes, Dematiaceae) on the growth of some deleterious fungi in poultry feed. AB - In previous work the authors have shown that some species of the Arthrinium genus are characterized by being able to produce secondary metabolites with antibiotic activity. The aim of this study was to evaluate the effect of raw extracts of the growth of three different Arthrinium strains against Aspergillus flavus, Aspergillus nidulans, Fusarium moniliforme and Penicillium purpurogenum when they were present in poultry feed. The results showed that the extracts reduced the growth of Aspergillus flavus and Fusarium moniliforme but could not inhibit the development of Aspergillus nidulans. Only the raw extract of A. aureum inhibited the growth of Penicillium purpurogenum. PMID- 11393748 TI - Detection of herpes simplex and varicella zoster viruses in clinical specimens using direct immunofluorescence and cell culture assays. AB - Patients (33 in toto) with a clinical diagnosis of herpes infections (simplex, zoster or chickenpox) were investigated for the presence of herpes simplex virus (HSV) and varicella zoster virus (VZV) in skin samples, using direct immunofluorescence and cell culture assays. Five patients with nonherpetic vesiculobullous disorders were included as negative controls. Of the 33 patients, nineteen (57.6%) were positive for HSV or VZV and fourteen (42.4%) were negative. Five controls were all negative for HSV or VZV. Of the nineteen positive patients, HSV was isolated from eight (42.1%) patients, by both direct immunofluorescence and cell culture assays. VZV was isolated from eleven (57.9%) patients, eleven (100%) by direct immunofluorescence assay, and six (54.5%) by cell culture assays. HSV was isolated from one patient clinically diagnosed as chickenpox (VZV), but otherwise the positive laboratory results were concordant with the clinical diagnosis. For epidemiological studies, atypical cases and immunocompromised patients the clinical diagnosis should be confirmed in the laboratory. PMID- 11393749 TI - Proflavine-mediated inactivation of Salmonella dublin exposed to visible sunlight in natural fresh water. AB - The survival of Salmonella dublin exposed to visible sunlight, and heterotrophic bacteria in freshwater microcosms in the presence and absence of the photosensitizer proflavine, was studied. Enumeration of S. dublin and the heterotrophic bacteria showed that in both illuminated and nonilluminated systems (without proflavine) the bacteria remained viable and culturable for at least 6 days. The optimal proflavine concentration (no effect in the dark and a maximal photoinactivation of salmonellae after irradiation) was 2 mg l(-1). In contrast to S. dublin, the heterotrophic bacteria overcame the initial inhibitory effect of proflavine. The possible use of photosterilization against contamination with pathogenic bacteria in water model ecosystems, is discussed. PMID- 11393750 TI - Effect of nitrite and propolis preservative on volatile basic nitrogen changes in meat products. AB - Pig meat was treated with 0.3% ethanol extracted propolis (EEP), 0.3% water extracted propolis (WEP), 0.4% dried residue of ethanol extracted propolis (DREEP) and 0.2% potassium sorbate (PS). The samples were stored for 8 weeks at 4, 10 and 20 degrees C, respectively. Volatile basic nitrogen (VBN) tests were carried out to measure the influence of temperature after 0, 2, 4 and 8 weeks. The VBN value in 0.3% EEP pork sausages was the lowest for all treatments after 8 weeks of storage at 4 degrees C. The 0.3% EEP treatment was superior to the 0.2% PS treatment. This was due to the lower VBN of the former than that of the latter. Hence, EEP, WEP and DREEP can serve as good chemical preservatives of pork meat products and can contribute to promote human health because they are naturally produced. PMID- 11393751 TI - Lactobacilli in human dental caries and saliva. AB - Samples (98 plaque and 72 saliva) from 93 patients with dental caries were investigated for Lactobacillus species which comprised 65 (62.5%) of 104 isolates. Yeasts (20.1%), Streptococcus spp. (8.7%), Staphylococcus spp. (2.9%) and a few unidentified species (5.8%), were also found. The Lactobacillus isolates were L. brevis (24.6%) L. fermentum (18.5%) L. casei (16.9%), L. delbrueckii (15.4%), L. plantarum (9.23%), L. acidophilus (7.69%), L. jensenii (4.62%), L. salivarius (1.54%) and L. gasseri (1.54%). The most common species was L. brevis (24.6%). The strains tested for beta-lactamase production showed 75.4% positive. All the Lactobacillus strains were tested for bacteriocin production against Escherichia coli, Salmonella spp., Shigella dysenteriae, S. sonnei, Klebsiella spp. and Campylobacter sp. All the lactobacilli except L. jensenii produced bacteriocin against at least one of the indicator organisms. The involvement of Lactobacillus in dental caries was established, although its role and mechanism is not well understood. The ability of Lactobacillus spp. to protect their host against certain diseases by inhibiting the growth of potential pathogens was evident. PMID- 11393752 TI - Candida albicans cell wall lytic enzyme produced by Streptomyces thermodiastaticus. AB - The production of lytic enzyme by Streptomyces thermodiastaticus was found to be affected by some growth conditions and nutritional factors. The highest enzyme production was obtained after 18 h of incubation at pH 5.5 and at 50 degrees C. The carbon source influenced the lytic enzyme production. A higher enzyme yield was obtained when Candida albicans cell wall (1 g/100 ml) was used as the sole carbon source. NaNO3 at 0.1 g/100 ml was the best nitrogen source for enzyme production. From all phosphorous sources, microelements, and growth factors tested, KH2PO4 (1 g/l), ZnSO4 (1 mg/I) and Tween 80 (0.1%), respectively, were found to favour the highest production of lytic enzymes by S. thermodiastaticus. The lytic enzymes mainly produced chitinolytic and proteolytic activities. PMID- 11393753 TI - Excitation functions of 125Te(p, xn)-reactions from their respective thresholds up to 100 MeV with special reference to the production of 124I. AB - Excitation functions of the nuclear reactions 125Te(p, xn) (119,120m, 120g, 121,122,123,124,125)I were measured for the first time from their respective thresholds up to 100 MeV using the stacked-foil technique. Thin samples were prepared by electrolytic deposition of 98.3% enriched 125Te on Ti-backing. In addition to experimental studies, excitation functions were calculated by the modified hybrid model code ALICE-IPPE. The experimental and theoretical data generally showed good agreement. From the measured cross section data, integral yields of (123,124,125)I were calculated. The energy range Ep 21 --> 15 MeV appears to be very suitable for the production of the medically interesting radionuclide 124I (T(1/2) = 4.18 d; I(beta)+ = 25%). The thick target yield of 124I amounts to 81 MBq/microA h and the level of 125I-impurity to 0.9%. The 125Te(p,2n)124I reaction gives 124I yield about four times higher than the commonly used 124Te(p,n)124I and 124Te(d,2n)124I reactions. The proposed production energy range is too high for small cyclotrons but large quantities of 124I can be produced with medium-sized commercial machines. PMID- 11393754 TI - Dry-distillation of astatine-211 from irradiated bismuth targets: a time-saving procedure with high recovery yields. AB - Astatine-211 was produced via the 209Bi(alpha,2n) 211At reaction. The radionuclide was isolated with a novel procedure employing dry-distillation of the irradiated target material. The astatine was condensed as a dry residue in a PEEK-capillary cryotrap. Distillation was completed within 1-2 min with isolation yields of 92 +/- 3%. Subsequent work-up of the nuclide resulted in final recovery yields of 79 +/- 3%. PMID- 11393755 TI - Determination of 241Am in sediments by isotope dilution high resolution inductively coupled plasma mass spectrometry (ID HR ICP-MS). AB - Trace levels (pg kg(-1)) of 241Am in sediments were determined by isotope dilution high resolution inductively coupled plasma mass spectrometry (ID HR ICP MS) using a microconcentric nebulizer. 241Am was isolated from major elements like Ca and Fe by different selective precipitations. In further steps. Am was first separated from other transuranic elements and purified by anion exchange and extraction chromatography prior to the mass spectrometric measurements. The ID HR ICP-MS results are compared with isotope dilution alpha spectrometry. PMID- 11393756 TI - Preparation of alpha-sources of U(VI) and Th(IV) by the electrodeposition technique in the presence of Ca2+ and some trivalent metals. AB - A new method for the electrodeposition of uranium and thorium from an aqueous/ethanol media in the presence of Ca(II) and some trivalent metals is described. The deposition is carried out at a constant weak voltage of 30 V and starting current intensity of 3 A. The deposition yields of uranium is investigated as a function of the amounts of various carriers and impurities such as Fe, Al, Bi, La and Ca as well as acetic, nitric and hydrochloric acids. All solutions containing up to 900 ppm of Ca2+ and 100-150 microg of the foreign ions namely: Fe, Bi or La can be used with no chemical separation. Uranium and thorium recoveries between 80 and 96% are obtained and the resolution is typically 25-40 keV. Procedure without chemical purification is proposed for a selective electrodeposition of U(VI) and Th(IV) from one another and from calcium. PMID- 11393757 TI - Geometrical effects on thermal neutron reflection of hydrogenous moderators using 241Am-Be source. AB - The variation of thermal reflection parameter with thicknesses of water and paraffin moderators was measured using an instrument consisting of 1 Ci 241Am-Be isotopic neutron source and 3He neutron detector. Based on experimental data obtained from two different source-detector-sample geometries, analytical expressions were derived for calculation of reflection parameters for various moderator thicknesses. The measured data for the two geometries exhibit saturation curves with saturation achieved at about 8 and 16 cm when sample container was placed at top and side of the source holder, respectively. By locating the source holder along the vertical axis of the sample container, measurements showed that the reflection parameter was very sensitive with the axial location and was strongly dependent on the moderator and its thickness. PMID- 11393758 TI - Technetium-99m labelling of glycosylated somatostatin-14. AB - This study presents a technetium-99m labelling method based on organometallic chemistry. It describes the simple mixing of a 99mTc(I)-carbonyl compound [99mTc(OH2)3(CO)3]+ with a histidine-tagged somatostatin-dextran (SMS-Dx-His) conjugate. Somatostatin and histidine was coupled to periodate activated dextran. The linkage was stabilised by reductive amination. The conjugate was then radiolabelled with 99mTc by using the 99mTc(CO)3 core. The labelling efficiency was 65-80% and the radiochemical purity > 95%. In the in vitro cysteine challenge, the result showed that 25% of the radiolabel was released after 1 h incubation at 37 degrees C (cysteine-conjugate at 1000:1 molar ratio). The radiolabelled SMS-Dx-His showed similar HPLC profile as the unlabelled conjugate. This labelling method, employing non reducing conditions, is useful for the labelling of peptides containing disulphide bonds. It should be possible to be used also for labelling with rhenium-188 for therapeutic applications. PMID- 11393759 TI - Dosimetric characteristics of 192Ir sources used in interstitial brachytherapy. AB - Dosimetric quantities of 192Ir seed (5 mm length) and wire (10 mm length) brachytherapy sources have been determined. The quantities were measured based on the protocol introduced by the Radiation Therapy Committee of the American Association of Physicists in Medicine (AAPM) Task Group 43. Quantities such as dose rate constant, (lambda), radial dose function, g(r), and anisotropy function, F(r, theta) were experimentally determined and the geometry function, G(r, theta), was calculated. TLD measurements were made in a polymethyl methacrylate (PMMA) phantom of dimensions 25 cm x 20 cm x 5 cm by means of LiF:Mg,Ti (TLD-100) dosimeters for distances of 1-10 cm for g(r), and the same distances at angles of 0-180 degrees for F(r, theta). Dose rate constant for 192Ir seed and wire were found to be 1.196+/-5 and 1.082+/-5% cGy h(-1) U(-1), respectively (1 U = unit of air Kerma strength = 1 microGy m2 h(-1) = 1 cGy cm2 h(-1)). The obtained results for g(r), G(r, theta) and F(r, theta) are also presented and discussed. PMID- 11393760 TI - Minimum detectable activity in in situ gamma-ray spectrometry. AB - The algorithm used to calculate a minimum detectable activity (A(D)) of an environmental volume sample measured by gamma-ray spectrometry was modified and adapted for in situ measurements performed at the standard 1 m height above ground. AD values of target radionuclides were determined from the in situ spectra collected at two disparate sites. A linear relationship between the two sets of A(D)s was found and the ratio between these two sets is equal to the square root of the ratio of the two respective absorbed dose rates measured at the sites at 1 m height. Absorbed dose rates were calculated using the concentration of potassium, thorium and uranium in the soils at the sites and found to agree well with the measured values. A(D) values can be predicted easily in in situ gamma spectrometry applying a simple experimental procedure that is based on the linear relationship. PMID- 11393761 TI - A new method for evaluating radon and thoron alpha-activities per unit volume inside and outside various natural material samples by calculating SSNTD detection efficiencies for the emitted alpha-particles and measuring the resulting track densities. AB - A Monte Carlo computer code for determining detection efficiencies of the CR-39 and LR-115 II solid-state nuclear track detectors (SSNTD) for alpha-particles emitted by the uranium and thorium series inside different natural material samples was developed. The influence of the alpha-particle initial energy on the SSNTD detection efficiencies was investigated. Radon (222Rn) and thoron (220Rn) alpha-activities per unit volume were evaluated inside and outside the natural material samples by exploiting data obtained for the detection efficiencies of the SSNTD utilized for the emitted alpha-particles, and measuring the resulting track densities. Results obtained were compared to those obtained by other methods. Radon emanation coefficients have been determined for some of the considered material samples. PMID- 11393762 TI - Contributions to the radiation chemistry of octasilsesquioxanes: unique traps for atomic hydrogen and free radicals at ambient temperature. AB - Evidence was given for trapping of H-atoms in fluid solutions of octasilsesquioxanes. Use was made of the positive effect of scavengers on the yield of trapped H-atoms. Carbon and silicon-centered radicals formed simultaneously through radiation chemical processes were detected. Most of the radical species were identified by the simulation of the corresponding ESR spectra. A wide range of thermal stability is covered by the radicals depending on the chemical nature of the substitutents of the cages and the precursors. PMID- 11393763 TI - A convenient method for discriminating between natural and depleted uranium by gamma-ray spectrometry. AB - A convenient method for discriminating between natural and depleted uranium reagent was developed by measuring and analyzing the gamma-ray spectra of some reagents with no standard source. The counting rates (R) of photoelectric peaks of gamma-rays from nuclides with the same radioactivity divided by their emission probability (B) are expressed as a function of gamma-ray energy. The radioactivities of 234Th and 234mPa and 21.72 times that of 235U are equal to the radioactivity of 235U in natural uranium. Therefore, the plot of 21.72-fold R/B for 235U should be on a curve fitted to the points for 234Th and 234mPa in natural uranium. Depleted uranium with a 235U isotopic composition of less than 0.68% could be discriminated from natural uranium in the case of a reagent containing 4.0 g of uranium. PMID- 11393764 TI - Sensitive measurement of positron emitters eluted from HPLC. AB - For sensitive analysis of the radioactive-metabolite in human PET, a radio-HPLC system coupled to a newly designed positron detector was constructed. The detector had the advantages of low noise level (1.7 +/- 1.0 cpm) and high sensitivity (32 +/- 1%) due to coincidence counting and large BGO crystals. Furthermore, the detector was easy to move, since a pair of the BGO housings coupled to photomultipliers was effectively arranged in parallel and a HPLC cell with different volume could be inserted between the BGO housing. This radio-HPLC system was useful for analyzing samples with low radioactivity. It was applied to the measurement of [11C]FLB457 in plasma, having high affinity and high selectivity with dopamine D2 receptors. Extremely low radioactivity of [11C]FLB457 (2500 dpm) could be analyzed by using the radio-HPLC system. The performance of this detector was compared with those of commercially available systems that had been used as sensitive detectors for HPLC. PMID- 11393765 TI - Determination of absorbed dose for photon and electron beams from a linear accelerator by comparing various protocols on different phantoms. AB - Protocols developed for high-energy dosimetry IAEA (Technical Reports Series No. 277, 1997), AAPM (Med. Phys. 10 (1983) 741: Med. Phys. 18 (1991) 73: Med. Phys. 21 (1994) 1251), IPEMB (Phys. Med. Biol. 41 (1996) 2557), and HPA (Phys. Med. Biol. 28 (1983) 1097) have continued to enhance precision in dose measurements and the optimization of radiotherapy procedures. While recent dosimetry protocols, including those due to the IAEA and IPEMB, have made a number of improvements compared with previous protocols, it is further desirable to develop absolute dosimetry methods of dose measurements. Measurements based on careful implementation of procedures contained within the various protocols have been carried out in an effort to determine the extent to which discrepancies exist among the protocols. Dose in water at dmax was measured using cylindrical and parallel-plate ionization chambers for 6 MV photon beams and 5 and 12 MeV electron beams. Results obtained from the use of the AAPM and HPA protocols for 6 MV photon beams were found to be 0.9% larger and 0.1% smaller, respectively, than those measured following the IAEA protocol. Calibration dose measurements for 5 and 12 MeV electron beams in water phantoms were found to agree to within 1%, this being well within recommendations from the ICRU and other sources regarding the accuracy of dose delivery. PMID- 11393766 TI - Analytical formulae for well-type NaI (Tl) and HPGe detectors efficiency computation. AB - A straightforward analytical formulae for the computation of total and full energy peak efficiencies of NaI (Tl) and HPGe well-type detectors are deduced. In addition, the attenuation of photons by the source container and the detector end cap materials is presented in a direct mathematical expression. Results are compared with previous treatments. PMID- 11393767 TI - ESR dosimetry using inorganic materials: a case study of Li2CO3 and CaSO4:Dy as prospective dosimeters. AB - The CO2- radical ion, detected by ESR technique in bones and teeth enamel, was proved to be invaluable in high level and retrospective dosimetry. In these matrices, impurity carbonate (at phosphate sites) was the precursor to CO2-. With a view to investigate the possibility of using inorganic materials such as lithium carbonate as ESR dosimeters, studies were carried out on gamma-irradiated Li2CO3. The intensity of radiation-induced ESR signals of Li2CO3 at g = 2.0036 (CO3-) and g = 2.0006 (CO2-) was followed as a function of gamma dose in the low dose range of 1-1350 Gy. It was observed that the intensity of the ESR signal at g = 2.0036 (CO3-) was in a linear relation with the radiation dose in the dose range 10-800 Gy and the signal at g = 2.0006 (CO2-) showed linear response in the dose range 5-800 Gy. The lowest dose that could be detected in the present studies using the signal of CO2- in Li2CO3 powder samples (approximately 50 mg) is 3.2 Gy. ESR studies were also carried out on the widely used TL dosimetric material CaSO4:Dy and in pure CaSO4 after gamma irradiation. The TL materials were used in powder as well as pellet forms. The linearity of ESR response with dose for powder and pellet forms of CaSO4: Dy was also studied using the signals at g = 2.0030 (SO3-) and at g = 2.0139 (SO4-). It was observed that the range of linearity of dose response extended between 20 and 1200 Gy, for SO3- signals. The results of dosimetric study indicate that the ESR-Li2CO3 system could be used in dosimetric applications in radiotherapy. However, for the actual applications further advancement is needed to lower the detection limit. The TL phosphor, CaSO4:Dy in powder and pellet forms, could be used as ESR dosimeter in the dose range 20-600 Gy. PMID- 11393768 TI - Gamma activity and geochemical features of building materials: estimation of gamma dose rate and indoor radon levels in Sicily. AB - A high-purity germanium detector has been used to measure the abundance of radium (Ra), thorium (Th) and potassium (K) in building materials used in Sicilian dwellings. The measurements were performed to evaluate which material was suitable for the construction of an enclosure, which would have a low background emission. The materials examined in this work showed concentrations of 226Ra, 232Th and 40K dramatically variable depending on the lithologies, particularly in the case of blocks, sands and aggregates commonly used in building materials in Sicily. The results are discussed and a criterion is indicated to reduce the radiation dose to humans. Since radon inlet is a major health problem, all the radiometric data and the geochemical features have been used to determine the radon exhalation, which arises from the disintegration of 226Ra in soils and walls of houses. From our experimental data it can be seen out that one of the geochemical parameters, Total Alkali (TA), may be an appropriate index to select materials of low radiological risk. PMID- 11393769 TI - Temporal variations of radon in soil related to earthquakes. AB - A radon detector with LR-115 nuclear track film was constructed for radon concentration measurements in soil. Temporal radon variations, as well as the barometric pressure, precipitation and temperature were measured for two years. Negative correlation between radon concentration in soil and barometric pressure was found. For some of the recorded earthquakes that occurred during the observation period, soil radon anomalies may be noticed one month before the quakes. PMID- 11393770 TI - Radon concentration in soil gas: a comparison of methods. AB - Various methods to determine 222Rn concentration in soil gas were tested at two sites with different soil types in a depth of about 1 m. They included instantaneous (spot), continuous (real-time) and time-averaging procedures. The first test site had a porous soil with high gravel content. At this site, radon concentrations obtained with the different methods agreed within the error limits of the procedures. The other site exhibited a largely impermeable clay soil and at this site, the measured radon concentrations differed up to a factor of two depending on the method. The applicability of the various procedures in the field is discussed. PMID- 11393771 TI - The application of thin layer activation for on-line erosion monitoring. AB - The washout of volcanic ash into the feedwater of a hydroelectric power station resulted in enhanced erosion of generator components. The damage prompted two novel applications of the thin layer activation (TLA) technique. TLA was used to select more resistant materials, and also as the basis of an automated, on-line alarm system to warn station operators on the likelihood of further damage. The latter application demonstrated that plant protection against irregular events is practical using TLA. PMID- 11393772 TI - Production and properties of beta-mannanase by free and immobilized cells of Aspergillus oryzae NRRL 3488. AB - Seven fungi were tested for production of mannanases. The highest mannanase activities were produced by Aspergillus oryzae NRRL 3488 after 7 days in static cultures. Mannanases were induced by gum locust bean (1.0%). The highest mannanase activity was produced when a mixture of peptone, urea and ammonium sulphate was used as nitrogen source. Zn2+ or Co2+ favoured enzyme production. The immobilized cells on Ca-alginate and agar were able to produce beta-mannanase for four runs with a slight decrease in the activity. The optimum temperature for enzyme reaction was 50-55 degrees C at pH 6.0. In the absence of substrate the enzyme was thermostable retaining 75% activity for 1 h at 50 degrees C, and 68% activity for 1 h at 60 degrees C. PMID- 11393773 TI - In vitro propagation and rhizome formation in Curcuma longa Linn. AB - Turmeric (Curcuma longa Linn.) which is cultivated by underground rhizomes is a slow propagating species. Multiplication and callus induction starting from the rhizome buds and shoot tips of C. longa in MS medium was carried out. A combination of naphthalene acetic acid (NAA; 1.0 mg/l) with kinetin (Kn; 1.0 mg/l) or NAA (1.0 mg/l) with 6-benzylaminopurine (BAP; 2.0 mg/l) was optimum for rapid clonal propagation of turmeric. A concentration of 2.5-3.0 mg/l of 2,4 dichlorophenoxy-acetic acid (2,4-D) was found to be optimum for callus induction. Regeneration of plantlets from a callus was successfully conducted in MS medium supplemented with standard growth hormones for multiplication at 25 +/- 2 degrees C under a 16 h photoperiod. These plantlets were successfully transferred to the field. Plantlets (4-month-old) were incubated in a medium containing different concentrations of sucrose supplemented with NAA (0.1 mg/l) and Kn (1.0 mg/l) at 27 +/- 2 degrees C under an 8 h photoperiod for induction of rhizomes. In vitro rhizome formation was observed in media containing 6 and 8% sucrose. PMID- 11393774 TI - Novel nitroimidazoles with trypanocidal and cell growth inhibition activities. AB - Chagas' disease, caused by the protozoan parasite Trypanosoma cruzi, affects 18 million people in Latin America and is an important cause of heart disease. Although transmission has been reduced, an effective therapy for the infected population is lacking. New nitroimidazoles were designed and synthesized aimed at the development of a trypanocidal drug. The coupling of nitroimidazoles with heterocyclic N-trifluoromethyltriazolyl and pyrazolyl groups, 5-[N-(3-(5 trifluoromethyl)-1H-1,2,4-triazolyl)]amino-1-methyl-4-nitroimidazole (compound 4) and 5-N-(1-pyrazolyl)-1-methyl-4-nitroimidazole (compound 5). The in vitro trypanocidal effects of compounds 4 and 5 were evaluated. The results demonstrated that compound 5 was the most active compound, killing about 100% and 64% of the parasites in 0.3 mg/ml and 0.003 mg/ml concentrations, respectively. Interestingly, compound 4 also inhibited myeloma cell growth in a dose-dependent manner. PMID- 11393775 TI - Cytogenetic studies on Choeroniscus minor (Chiroptera, Phyllostomidae) from the Amazon region. AB - The Choeroniscus genus (Glossophaginae, Phyllostomidae) has five monotypic species: C. minor, C. godmani, C. intermedius, C. inca and C. periosus. This paper analyses the karyotype of a female C. minor, collected close to the Guama river (Belem, Para, Brazil). G-, C-banding and NOR-staining were performed. This species has 2n = 20 chromosomes, where there are two bi-armed pairs (numbers 1 and 9) and seven subtelocentric pairs (2, 3, 4, 5, 6, 7 and 8). The probable X chromosome is a submetacentric. The constitutive heterochromatin can be found in the short arm of five subtelocentric pairs (4, 5, 6, 7 and 8) and is centromeric in the bi-armed pairs numbers 1 and 9, and the X chromosome. The heterochromatic bands are heteromorphic in three pairs (1, 2 and 3). Active NOR were observed in the short arms of eight subtelocentric chromosomes, suggesting that at least four pairs are nucleolar organizers. This paper describes for the first time the karyotype of C. minor from the Amazon region. PMID- 11393776 TI - Cytogenetic characteristics of species of the Chironomus plumosus group (Chironomidae, Diptera) in Finland. AB - The cytogenetic characteristics of species of the Chironomus plumosus group in Finland were examined. The species included C. balatonicus Devai, Wulker, Scholl, C. entis Shobanov, C. plumosus L., C. muratensis Ryser, Scholl, Wulker, and two karyotypes of unknown species. All belonged to the thummi complex having 2n = 8 chromosomes, with arm combinations of AB, CD, EF, G. In C. balatonicus arms A to G the band sequences corresponded with those of other populations. A new band sequence was found in arm B. In C. entis an arm A had a band sequence similar to those in C. agilis. A large pericentric inversion was observed in chromosome AB. The arms B to G had band sequences typical for C. entis. The chromosome arms A and B in C. plumosus from Lake Marsjon had band sequences corresponding to those of C. agilis and C. entis, respectively. C. plumosus from Helsinki may be a more divergent population with a large amount of centromere heterochromatin. C. muratensis was not distinguishable by band sequences from those of other Palaearctic populations. Two new karyotypes similar to those of species of the plumosus group have been described. PMID- 11393777 TI - Expression of osteoprotegerin, receptor activator of NF-kappaB ligand (osteoprotegerin ligand) and related proinflammatory cytokines during fracture healing. AB - Fracture healing is a unique biological process regulated by a complex array of signaling molecules and proinflammatory cytokines. Recent evidence for the role of tumor necrosis family members in the coupling of cellular functions during skeletal homeostasis suggests that they also may be involved in the regulation of skeletal repair. The expression of a number of cytokines and receptors that are of functional importance to bone remodeling (osteoprotegerin [OPG], macrophage colony-stimulating factor [M-CSF], and osteoprotegerin ligand [receptor activator of NF-kappaB ligand (RANKL)]), as well as inflammation (tumor necrosis factor alpha [TNF-alpha] and its receptors, and interleukin-1alpha [IL-1alpha] and -beta and their receptors) were analyzed over a 28-day period after the generation of simple transverse fractures in mouse tibias. OPG was expressed constitutively in unfractured bones and elevated levels of expression were detected throughout the repair process. It showed two distinct peaks of expression: the first occurring within 24 h after fracture and the second at the time of peak cartilage formation on day 7. In contrast, the expression of RANKL was nearly undetectable in unfractured bones but strongly induced throughout the period of fracture healing. The peak in expression of RANKL did not correlate with that of OPG, because maximal levels of expression were seen on day 3 and day 14, when OPG levels were decreasing. M-CSF expression followed the temporal profile of RANKL but was expressed at relatively high basal levels in unfractured bones. TNF-alpha, lymphotoxin-beta (LT-beta), IL-1alpha, and IL-1beta showed peaks in expression within the first 24 h after fracture, depressed levels during the period of cartilage formation, and increased levels of expression on day 21 and day 28 when bone remodeling was initiated. Both TNF-alpha receptors (p55 and p75) and the IL 1RII receptor showed identical patterns of expression to their ligands, while the IL-1R1 was expressed only during the initial period of inflammation on day 1 and day 3 postfracture. Both TNF-alpha and IL-1alpha expression were localized primarily in macrophages and inflammatory cells during the early periods of inflammation and seen in mesenchymal and osteoblastic cells later during healing. TNF-alpha expression also was detected at very high levels in hypertrophic chondrocytes. These data imply that the expression profiles for OPG, RANKL, and M CSF are tightly coupled during fracture healing and involved in the regulation of both endochondral resorption and bone remodeling. TNF-alpha and IL-1 are expressed at both very early and late phases in the repair process, which suggests that these cytokines are important in the initiation of the repair process and play important functional roles in intramembraneous bone formation and trabecular bone remodeling. PMID- 11393778 TI - The ratio of messenger RNA levels of receptor activator of nuclear factor kappaB ligand to osteoprotegerin correlates with bone remodeling indices in normal human cancellous bone but not in osteoarthritis. AB - The determinants of cancellous bone turnover and trabecular structure are not understood in normal bone or skeletal disease. Bone remodeling is initiated by osteoclastic resorption followed by osteoblastic formation of new bone. Receptor activator of nuclear factor kappaB ligand (RANKL) is a newly described regulator of osteoclast formation and function, the activity of which appears to be a balance between interaction with its receptor RANK and with an antagonist binding protein osteoprotegerin (OPG). Therefore, we have examined the relationship between the expression of RANKL, RANK, and OPG and indices of bone structure and turnover in human cancellous bone from the proximal femur. Bone samples were obtained from individuals with osteoarthritis (OA) at joint replacement surgery and from autopsy controls. Histomorphometric analysis of these samples showed that eroded surface (ES/BS) and osteoid surface (OS/BS) were positively associated in both control (p < 0.001) and OA (p < 0.02), indicating that the processes of bone resorption and bone formation remain coupled in OA, as they are in controls. RANKL, OPG, and RANK messenger RNA (mRNA) were abundant in human cancellous bone, with significant differences between control and OA individuals. In coplotting the molecular and histomorphometric data, strong associations were found between the ratio of RANKL/OPG mRNA and the indices of bone turnover (RANKL/OPG vs. ES/BS: r = 0.93, p < 0.001; RANKL/OPG vs. OS/BS: r = 0.80, p < 0.001). These relationships were not evident in trabecular bone from severe OA, suggesting that bone turnover may be regulated differently in this disease. We propose that the effective concentration of RANKL is related causally to bone turnover. PMID- 11393779 TI - The Src signaling pathway regulates osteoclast lysosomal enzyme secretion and is rapidly modulated by estrogen. AB - To investigate the role of the pp60src signaling pathway in osteoclast activity, we have used dominant negative pp60src, c-ras, and c-raf expression vectors to individually disrupt their functions in osteoclasts. Osteoclasts were transiently transfected and secretions of cathepsin B/K and tartrate-resistant acid phosphatase (TRAP) were monitored. Expression of these constructs increased secretion of lysosomal enzymes. In contrast, constitutively active pp60src expression caused decreased lysosomal enzyme secretion. Osteoclasts respond to 17 beta estradiol (17betaE2) treatment with decreased lysosomal enzyme secretion. Therefore, we investigated the effects of E2 on pp60src kinase activity and observed an E2 time- and dose-dependent decrease in cytoskeletal membrane associated pp60src tyrosine kinase activity. We have shown that estrogen decreases lysosomal enzyme gene expression and secretion; so we have examined the effects of the expression constructs on estrogen regulation of enzyme secretion. Constitutively active pp60src blocked E2 effects on secretion whereas expression of dominant negative pp60src, c-Ras, or c-Raf enhanced E2 effects. These data support that the kinase domain of cytoskeletal-associated pp60src is likely to be involved in the regulation of lysosomal enzyme secretion. PMID- 11393780 TI - Modulation of 11beta-hydroxysteroid dehydrogenase isozymes by proinflammatory cytokines in osteoblasts: an autocrine switch from glucocorticoid inactivation to activation. AB - Tissue damage by proinflammatory cytokines is attenuated at both systemic and cellular levels by counter anti-inflammatory factors such as corticosteroids. Target cell responses to corticosteroids are dependent on several factors including prereceptor regulation via local steroidogenic enzymes. In particular, two isozymes of 11beta-hydroxysteroid dehydrogenase (11beta-HSD), by interconverting hormonally active cortisol (F) to inactive cortisone (E), regulate the peripheral action of corticosteroids 11beta-HSD1 by converting E to F and 11beta-HSD2 by inactivating F to E. In different in vitro and in vivo systems both 11beta-HSD isozymes have been shown to be expressed in osteoblasts (OBs). Using the MG-63 human osteosarcoma cell-line and primary cultures of human OBs, we have studied the regulation of osteoblastic 11beta-HSD isozyme expression and activity by cytokines and hormones with established roles in bone physiology. In MG-63 cells, interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF alpha) potently inhibited 11beta-HSD2 activity (cortisol-cortisone conversion) and messenger RNA (mRNA) levels in a dose-dependent manner while stimulating reciprocal expression of 11beta-HSD1 mRNA and activity (cortisone-cortisol conversion). A similar rise in 11beta-HSD1 reductase activity also was observed in primary cultures of OBs treated with 10 ng/ml TNF-alpha. Pretreatment of MG-63 cells with 0.1 ng/ml IL-1beta resulted in increased cellular sensitivity to physiological glucocorticoids as shown by induction of serum and glucocorticoid inducible kinase (SGK; relative increase with 50 nM F but no IL-1beta pretreatment 1.12 +/- 0.34; with pretreatment 2.63 +/- 0.50; p < 0.01). These results highlight a novel mechanism within bone cells whereby inflammatory cytokines cause an autocrine switch in intracellular corticosteroid metabolism by disabling glucocorticoid inactivation (11beta-HSD2) while inducing glucocorticoid activation (11beta-HSD1). Therefore, it can be postulated that some of the effects of proinflammatory cytokines within bone (e.g., periarticular erosions in inflammatory arthritis) are mediated by this mechanism. PMID- 11393781 TI - Mechanical strain and estrogen activate estrogen receptor alpha in bone cells. AB - Bone cells' early responses to estrogen and mechanical strain were investigated in the ROS 17/2.8 cell line. Immunoblotting with antiphosphorylated estrogen receptor a (ER-alpha) antibody showed that when these cells were exposed for 10 minutes to estrogen (10(-8) M) or a single period of cyclic dynamic strain (peak 3400 microepsilon, 1 Hz, 600 cycles), there was an increase in the intensity of a 66-kDa band, indicating phosphorylation of ser122 in the amino terminus of ER alpha. Increased phosphorylation was detected within 5 minutes of exposure to estrogen and 5 minutes after the end of the period of strain. Estrogen and strain also activated the mitogen-activated protein kinase (MAPK) family member extracellular regulated kinase-1 (ERK-1). Increases in ERK activation coincided with increased ER-alpha phosphorylation. Activation of ERK-1 and the phosphorylation of ER-alpha, by both estrogen and strain, were prevented by the MAP kinase kinase (MEK) inhibitor U0126 and the protein kinase A (PKA) inhibitor (PKI). These data support previous suggestions that resident bone cells' early responses to strain and estrogen share a common pathway, which involves ER-alpha. This pathway also appears to involve PKA and ERK-mediated phosphorylation of ser122 within the amino terminus of ER-alpha. Reduced availability of this pathway when estrogen levels are reduced could explain diminished effectiveness of mechanically related control of bone architecture after the menopause. PMID- 11393782 TI - Monitoring of periprosthetic BMD after uncemented total hip arthroplasty with dual-energy X-ray absorptiometry--a 3-year follow-up study. AB - Insertion of a metallic implant into the femur changes bone loading conditions and results in remodeling of femoral bone. To quantify changes in bone mass after uncemented total hip arthroplasty (THA), we monitored femoral bone with dual energy X-ray absorptiometry (DXA). The periprosthetic bone mineral density (BMD) was measured with Lunar DPX densitometry in seven Gruen zones and the total periprosthetic area at scheduled time intervals in 22 patients during a 3-year follow-up. BMD decreased significantly almost in all Gruen zones during the first 3 months, ranging from 3.4% to 14.4% (p < 0.05 top < 0.001). At the end of the first year, the most remarkable decrease in BMD was found in the calcar (zone 7; 22.9%). During the second postoperative year, a slight restoration of periprosthetic bone mass was recorded. During the third year, no significant changes in BMD were found. The preoperative BMD was the only factor that was significantly related to the periprosthetic bone loss. Clearly, the early periprosthetic bone loss noticed during the 3 months after THA is caused by mainly limited weight bearing to the operated hip and stress shielding. We suggest that the restoration of bone mass is a sign of successful osteointegration between bone and metallic implant. DXA is a suitable tool to follow the bone response to prosthetization and will increase our knowledge on the behavior of bone after THA. PMID- 11393783 TI - Role of D1 and E cyclins in cell cycle progression of human fibroblasts adhering to cementum attachment protein. AB - Cementum attachment protein (CAP) is a collagenous protein present in the matrix of tooth cementum that mediates preferential attachment of some mesenchymal cell types, and CAP binding capacity is related to mineralizing tissue-forming capacity in culture. We have examined if adhesion to surfaces containing CAP as the only attachment protein permits human fibroblasts to escape G1 arrest and synthesize DNA, and if adhesion to CAP modulates the levels of cyclins D1 and E. Human gingival fibroblasts (HGFs) were serum-starved, trypsinized, and added to plates coated with CAP or bovine serum albumin (BSA). Cells were then exposed to either 10% fetal bovine serum (FBS) or to cementum-derived growth factor (CGF), an insulin-like growth factor I (IGF-I)-like molecule sequestered in tooth cementum, plus epidermal growth factor (EGF). DNA synthesis was measured as [3H]thymidine uptake, and cyclin D1 and E levels were determined by Western analysis. Cyclin E-dependent kinase (Cdk) activity was assessed in terms of H1 kinase activity in immunoprecipitates of cyclin E. Cells adhering to CAP synthesized DNA, whereas on BSA they remained unattached and did not synthesize DNA. Protein levels of cyclin D1 were higher in cells adhering to CAP in the absence and presence of growth factors. Cyclin E levels were not affected by adhesion alone, but they increased in the presence of growth factors. Cyclin E associated kinase activity was higher in cells adherent on CAP, and it increased further in the presence of growth factors. Our results indicate that adhesion to CAP increases cyclin D1 levels and cyclin E-associated Cdk activity, and that these increases contribute to cell cycle progression. We previously observed that the signaling reactions induced during adhesion are characteristic of the CAP; together these observations indicate that specific matrix components present in the local environment can contribute to recruitment and differentiation of specific cell types for normal homeostasis and wound healing. PMID- 11393784 TI - Growth hormone induces bone morphogenetic proteins and bone-related proteins in the developing rat periodontium. AB - The hypothesis that growth hormone (GH) up-regulates the expression of enzymes, matrix proteins, and differentiation markers involved in mineralization of tooth and bone matrices was tested by the treatment of Lewis dwarf rats with GH over 5 days. The molar teeth and associated alveolar bone were processed for immunohistochemical demonstration of bone morphogenetic proteins 2 and 4 (BMP-2 and -4), bone morphogenetic protein type IA receptor (BMPR-IA), bone alkaline phosphatase (ALP), osteocalcin (OC), osteopontin (OPN), bone sialoprotein (BSP), and E11 protein (E11). The cementoblasts, osteoblasts, and periodontal ligament (PDL) cells responded to GH by expressing BMP-2 and -4, BMPR-IA, ALP, OC, and OPN and increasing the numbers of these cells. No changes were found in patterns of expression of the late differentiation markers BSP and E11 in response to GH. Thus, GH evokes expression of bone markers of early differentiation in cementoblasts, PDL cells, and osteoblasts of the periodontium. We propose that the induction of BMP-2 and -4 and their receptor by GH compliments the role of GH induced insulin-like growth factor 1 (IGF-1) in promoting bone and tooth root formation. PMID- 11393785 TI - Monitoring metastatic behavior of human tumor cells in mice with species-specific polymerase chain reaction: elevated expression of angiogenesis and bone resorption stimulators by breast cancer in bone metastases. AB - Tumor-stroma interactions are of primary importance in determining the pathogenesis of metastasis. Here, we describe the application of sensitive competitive polymerase chain reaction (PCR) techniques for detection and quantitation of human breast cancer cells (MDA-MB-231) in an in vivo mouse model of experimental metastasis. Human-specific oligonucleotide primers in competitive PCR reactions were used to quantify the amount of MDA-MB-231 cells per tissue per organ. Using this species-specific (semi)quantitative PCR approach, gene expression patterns of (human) tumor cells or (mouse) stromal cells in metastatic lesions in the skeleton or soft tissues were investigated and compared. In all metastatic lesions, MDA-MB-231 cells express angiogenic factors (vascular endothelial growth factors [VEGFs]; VEGF-A, -B, and -C) and bone-acting cytokines (parathyroid hormone-related protein [PTHrP] and macrophage colony-stimulating factor [M-CSF]). In these metastases, PECAM-1-positive blood vessels and stromal cells of mouse origin are detected. The latter express angiogenic factors and markers of sprouting vessels (VEGF receptors flt-1/flk - 1/flk-4 and CD31/PECAM 1). Strikingly, steady-state messenger RNA (mRNA) levels of VEGF-A and -B and the major bone resorption stimulators PTHrP and M-CSF by tumor cells were elevated significantly in bone versus soft tissues (p < or = 0.05, p < or = 0.0001, p < or = 0.001, and p < or = 0.05, respectively), indicating tissue-specific expression of these tumor progression factors. In conclusion, MDA-MB-231 breast cancer cells express a variety of factors in vivo that have been implicated in metastatic bone disease and that correlate with poor survival of patients with breast cancer. We hypothesize that the observed up-regulated expression of angiogenic and bone resorbing factors by the breast cancer cells in the skeleton underlie the clinically observed osteotropism of breast cancer cells and pathogenesis of osteolytic bone metastases. The application of the species-specific competitive PCR-based assay in vivo can provide new information concerning the involvement of gene families in tumor progression and metastatic disease and greatly facilitates the study of tumor-stroma interactions in cancer invasion and metastasis. PMID- 11393786 TI - Histomorphometry of the embryonic avian growth plate by proton nuclear magnetic resonance microscopy. AB - Quantitative nuclear magnetic resonance (NMR) microscopy was used to characterize the biochemical and morphological properties of the different zones within the growth plate of an embryonic chick femur. For precalcified tissue, water proton transverse relaxation times (T2) and magnetization transfer values (MT) were directly and inversely dependent, respectively, on tissue cellularity, defined as the intracellular area per unit area on histological sections. T2 values extrapolated for intra- and extracellular water were 96 ms and 46 ms, respectively. The extracellular T2 was comparable with that measured for mature cartilage. The MT values extrapolated for intra- and extracellular compartments were 0.32 and 0.85, respectively. These values were comparable with those values reported in the literature for cell pellets and for mature cartilage tissue. Thus, cellularity dominated the NMR properties of this immature cartilage tissue. Mineral deposits within calcified cartilage and periosteal bone invoked NMR relaxation processes that were dependent on the inorganic mineral phase. Additionally, collagen molecules present in mineralized zones gave rise to a significant MT effect. These results show the utility of water proton NMR microscopy for assessing both the organic and inorganic phases within mineralized tissues. PMID- 11393787 TI - The contribution of reduced peak accrual of bone and age-related bone loss to osteoporosis at the spine and hip: insights from the daughters of women with vertebral or hip fractures. AB - The genetic hypothesis states that a daughter will resemble her mother by about 50% in a given trait because she shares, on average, half her genes. We used this trait resemblance in mothers and daughters to determine whether abnormalities in volumetric bone mineral density (vBMD) or bone size in women with fractures originate in growth or aging. vBMD and volume of the third lumbar vertebra and femoral neck were estimated using posteroanterior (PA) scanning by dual-energy X ray absorptiometry (DXA). Vertebral volume was estimated as (scan area)(3/2) and femoral neck volume was pi* (width/2)(2)* height. vBMD was bone mineral content (BMC)/volume. The data were expressed as age-specific SD or Z scores (mean +/- SEM). Vertebral vBMD was reduced by -0.98 +/- 0.14 SD (p < 0.001) in 34 women with vertebral fractures, and by -0.36 +/- 0.13 SD (p < 0.05) in their 44 premenopausal daughters. The vBMD deficit in the daughters (relative to age matched controls) was no different from one-half their mothers' deficit (relative to their age-matched controls). Vertebral volume was reduced in the women with vertebral fractures relative to age-matched controls (-0.77 +/- 0.15 SD; p < 0.001), but not in their daughters (-0.17 +/- 0.13 SD, NS). The 31 women with hip fractures and their 41 premenopausal daughters had no deficits in vertebral volume or vBMD. Femoral neck vBMD was reduced in the women with hip fractures ( 1.24 +/- 0.12 SD; p < 0.001) but not in their daughters (-0.17 +/- 0.13 SD, NS). Femoral neck volume was increased by 0.98 +/- 0.30 SD (p < 0.05) in women with hip fractures (relative to age-matched controls) and by 0.54 +/- 0.14 SD (p < 0.001) in their daughters (relative to age-matched controls); that is, about one half that of their mothers. We propose that women with vertebral fractures have reduced vertebral vBMD because of, in large part, reduced accrual of bone during growth (because the deficit in their daughters was almost one-half their mothers' deficit); reduced vertebral volume in women with vertebral fractures is caused by reduced periosteal apposition during aging (because their daughters have no deficit in vertebral volume). Women with hip fractures have reduced vBMD because of age-related bone loss (because their daughters have no deficit in vBMD) but the increased femoral neck volume is growth related (because their daughters' femoral neck size is increased by one-half as much). The pathogenesis of bone fragility at the axial and appendicular skeleton is heterogeneous and has its origins in growth and aging. PMID- 11393788 TI - Structural adaptation to changing skeletal load in the progression toward hip fragility: the study of osteoporotic fractures. AB - Longitudinal, dual-energy X-ray absorptiometry (DXA) hip data from 4187 mostly white, elderly women from the Study of Osteoporotic Fractures were studied with a structural analysis program. Cross-sectional geometry and bone mineral density (BMD) were measured in narrow regions across the femoral neck and proximal shaft We hypothesized that altered skeletal load should stimulate adaptive increases or decreases in the section modulus (bending strength index) and that dimensional details would provide insight into hip fragility. Weight change in the approximately 35 years between scan time points was used as the primary indicator of altered skeletal load. "Static" weight was defined as within 5% of baseline weight, whereas "gain" and 'loss" were those who gained or lost >5%, respectively. In addition, we used a frailty index to better identify those subjects undergoing changing in skeletal loading. Subjects were classified as frail if unable to rise from a chair five times without using arm support. Subjects who were both frail and lost weight (reduced loading) were compared with those who were not frail and either maintained weight (unchanged loading) or gained weight (increased loading). Sixty percent of subjects (n = 2,559) with unchanged loads lost BMD at the neck but not at the shaft, while section moduli increased slightly at both regions. Subjects with increasing load (n = 580) lost neck BMD but gained shaft BMD; section moduli increased markedly at both locations. Those with declining skeletal loads (n = 105) showed the greatest loss of BMD at both neck and shaft; loss at the neck was caused by both increased loss of bone mass and greater subperiosteal expansion; loss in shaft BMD decline was only caused by greater loss of bone mass. This group also showed significant declines in section modulus at both sites. These results support the contention that mechanical homeostasis in the hip is evident in section moduli but not in bone mass or density. The adaptive response to declining skeletal loads, with greater rates of subperiosteal expansion and cortical thinning, may increase fragility beyond that expected from the reduction in section modulus or bone mass alone. PMID- 11393790 TI - Bone mineral density and bone loss measured at the radius to predict the risk of nonspinal osteoporotic fracture. AB - Low bone mineral density (BMD) and, probably, the rate of bone loss (RBL) are associated with the risk of osteoporotic fractures. To estimate the risk of nonspinal fracture in osteoporotic women, we measured BMD and RBL in a prospective study (average follow-up, 5.38 years) in 656 postmenopausal women. The women were considered in three groups: group A (whole population), group B (women under the age of 65 years) and group C (women over the age of 65 years). At the beginning of the study, BMD was measured at the distal radius (DR) and at the proximal radius (PR) using a single-energy densitometer. BMD measurements made 2 years previously in the same patients were used to calculate RBL. Then patients were checked annually for nonspine fracture due to minor trauma. During follow-up, 121 nonspinal fractures were detected. Women with fractures were older and had lower BMD. With the Cox regression, age-corrected BMD at both DR and PR predicts fracture risk in groups A and B but not in group C. After correction for potential confounders, DR still predicts fractures in groups A and B whereas PR predicts fractures only in group B. In group C, only the RBL at the PR was predictive of the fracture risk as well as in the other two groups. Specific types of fractures are predictable in the whole population at the wrist. In conclusion, radial BMD predicts the risk of nonspine fractures except in women over the age of 65 years. The RBL at the PR is an effective predictor of fracture risk also in women over the age of 65 years. PMID- 11393789 TI - Number and proliferative capacity of osteogenic stem cells are maintained during aging and in patients with osteoporosis. AB - Decreased bone formation is an important pathophysiological mechanism responsible for bone loss associated with aging and osteoporosis. Osteoblasts (OBs), originate from mesenchymal stem cells (MSCs) that are present in the bone marrow and form colonies (termed colony-forming units-fibroblastic [CFU-Fs]) when cultured in vitro. To examine the effect of aging and osteoporosis on the MSC population, we quantified the number of MSCs and their proliferative capacity in vitro. Fifty-one individuals were studied: 38 normal volunteers (23 young individuals [age, 22-44 years] and 15 old individuals [age, 66-74 years]) and 13 patients with osteoporosis (age, 58-83 years). Bone marrow was aspirated from iliac crest; mononuclear cells were enriched in MSCs by magnetic activated cell sorting (MACS) using STRO-1 antibody. Total CFU-F number, size distribution, cell density per CFU-F, number of alkaline phosphatase positive (ALP+) CFU-Fs, and the total ALP+ cells were determined. In addition, matrix mineralization as estimated by alizarin red S (AR-S) staining was quantified. No significant difference in colony-forming efficiency between young individuals (mean +/- SEM; 87 +/- 12 CFU Fs/culture), old individuals (99 +/- 19 CFU-Fs/culture), and patients with osteoporosis (129 +/- 13 CFU-Fs/culture; p = 0.20) was found. Average CFU-F size and cell density per colony were similar in the three groups. Neither the percentage of ALP+ CFU-Fs (66 +/- 6%, 65 +/- 7%, and 72 +/- 4% for young individuals, old individuals, and patients with osteoporosis, respectively) nor the percentage of ALP+ cells per culture (34 +/- 5%, 40 +/- 6%, and 41 +/- 4%) differed between groups. Finally, mineralized matrix formation was similar in young individuals, old individuals, and patients with osteoporosis. Our study shows that the number and proliferative capacity of osteoprogenitor cells are maintained during aging and in patients with osteoporosis and that other mechanisms must be responsible for the defective osteoblast (OB) functions observed in these conditions. PMID- 11393791 TI - Characterization of mouse Atp6i gene, the gene promoter, and the gene expression. AB - Solubilization of bone mineral by osteoclasts depends on the formation of an acidic extracellular compartment through the action of a V-type ATPase. We previously cloned a gene encoding a putative osteoclast-specific proton pump subunit, termed OC-116 kDa, approved mouse Atp6i (ATPase, H+ transporting, [vacuolar proton pump] member I). The function of Atp6i as osteoclast-specific proton pump subunit was confirmed in our mouse knockout study. However, the transcription regulation of Atp6i remains largely unknown. In this study, the gene encoding mouse Atp6i and the promoter have been isolated and completely sequenced. In addition, the temporal and spatial expressions of Atp6i have been characterized. Intrachromosomal mapping studies revealed that the gene contains 20 exons and 19 introns spanning approximately 11 kilobases (kb) of genomic DNA. Alignment of the mouse Atp6i gene exon sequence and predicted amino acid sequence to that of the human reveals a strong homology at both the nucleotide (82%) and the amino acid (80%) levels. Primer extension assay indicates that there is one transcription start site at 48 base pairs (bp) upstream of the initiator Met codon. Analysis of 4 kb of the putative promoter region indicates that this gene lacks canonical TATA and CAAT boxes and contains multiple putative transcription regulatory elements. Northern blot analysis of RNAs from a number of mouse tissues reveals that Atp6i is expressed predominantly in osteoclasts, and this predominant expression was confirmed by reverse-transcription polymerase chain reaction (RT-PCR) assay and immunohistochemical analysis. Whole-mount in situ hybridization shows that Atp6i expression is detected initially in the headfold region and posterior region in the somite stage of mouse embryonic development (E8.5) and becomes progressively restricted to anterior regions and the limb bud by E9.5. The expression level of Atp6i is largely reduced after E10.5. This is the first report of the characterizations of Atp6i gene, its promoter, and its gene expression patterns during mouse development. This study may provide valuable insights into the function of Atp6i, its osteoclast-selective expression, regulation, and the molecular mechanisms responsible for osteoclast activation. PMID- 11393792 TI - Canine COL1A2 mutation resulting in C-terminal truncation of pro-alpha2(I) and severe osteogenesis imperfecta. AB - RNA and type I collagen were analyzed from cultured skin fibroblasts of a Beagle puppy with fractures consistent with type III osteogenesis imperfecta (OI). In a nonisotopic RNAse cleavage assay (NIRCA), the proband's RNA had a unique cleavage pattern in the region of COL1A2 encoding the C-propeptide. DNA sequence analyses identified a mutation in which nucleotides 3991-3994 ("CTAG") were replaced with "TGTCATTGG." The first seven bases of the inserted sequence were identical to nucleotides 4002-4008 of the normal canine COL1A2 sequence. The resulting frameshift changed 30 amino acids and introduced a premature stop codon. Reverse transcription polymerase chain reaction (RT-PCR) with primers flanking the mutation site amplified two complementary DNA (cDNA) fragments for the proband and a single product for the control. Restriction enzyme digestions also were consistent with a heterozygous mutation in the proband. Type I procollagen labeled with [3H]proline was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Increased density of pC-alpha2(I) suggested comigration with the similarly sized pro-alpha2(I) derived from the mutant allele. Furthermore, a-chains were overhydroxylated and the ratio of alpha1(I):alpha2(I) was 3.2:1, consistent with the presence of alpha1(I) homotrimers. Analyses of COL1A2 and type I collagen were both consistent with the described heterozygous mutation affecting the pro-alpha2(I) C-propeptide and confirmed a diagnosis of OI. PMID- 11393793 TI - Intraosseous meningioma--a mimicry of Paget's disease? PMID- 11393794 TI - Osteoblast apoptosis and bone turnover. AB - With the discoveries of different death mechanisms, an emerging definition of apoptosis is the process of cell death associated with caspase activation or caspase-mediated cell death. This definition accepts that caspases represent the final common mechanistic pathway in apoptosis. Apoptosis may be triggered either by activation events that target mitochondria or endoplasmic reticulum or by activation of cell surface "death receptors," for example, those in the tumor necrosis factor (TNF) superfamily. In the postnatal and adult skeleton, apoptosis is integral to physiological bone turnover, repair, and regeneration. The balance of osteoblast proliferation, differentiation, and apoptosis determines the size of the osteoblast population at any given time. Although apoptosis has been recorded in many studies of bone, the selective mechanisms invoked in the different models studied rarely have been identified. This review offers a broad overview of the current general concepts and controversies in apoptosis research and then considers specific examples of osteoblast apoptosis pertinent to skeletal development and to the regulation of bone turnover. In reviewing selected work on interdigital apoptosis in the developing skeleton, we discuss the putative roles of the bone morphogenetic proteins (BMPs), Msx2, RAR-gamma, and death inducer obliterator 1 (DIO-1). In reviewing factors regulating apoptosis in the postnatal skeleton, we discuss roles of cytokines, growth factors, members of the TNF pathway, and the extracellular matrix (ECM). Finally, the paradoxical effects of parathyroid hormone (PTH) on osteoblast apoptosis in vivo are considered in the perspective of a recent hypothesis speculating that this may be a key mechanism to explain the anabolic effects of the hormone. An improved understanding of the apoptotic pathways and their functional outcomes in bone turnover and fracture healing may facilitate development of more targeted therapeutics to control bone balance in patients with osteoporosis and other skeletal diseases. PMID- 11393795 TI - Genome screen for quantitative trait loci underlying normal variation in femoral structure. AB - Femoral structure contributes to bone strength at the proximal femur and predicts hip fracture risk independently of bone mass. Quantitative components of femoral structure are highly heritable traits. To identify genetic loci underlying variation in these structural phenotypes, we conducted an autosomal genome screen in 309 white sister pairs. Seven structural variables were measured from femoral radiographs and used in multipoint sib-pair linkage analyses. Three chromosomal regions were identified with significant evidence of linkage (log10 of the odds ratio [LOD] > 3.6) to at least one femoral structure phenotype. The maximum LOD score of 4.3 was obtained for femur neck axis length on chromosome 5q. Evidence of linkage to chromosome 4q was found with both femur neck axis length (LOD = 3.9) and midfemur width (LOD = 3.5). Significant evidence of linkage also was found to chromosome 17q, with a LOD score of 3.6 for femur head width. Two additional chromosomal regions 3q and 19p gave suggestive (LOD > 2.2) evidence of linkage with at least two of the structure phenotypes. Chromosome 3 showed evidence of linkage with pelvic axis length (LOD = 3.1), midfemur width (LOD = 2.8), and femur head width (LOD = 2.3), spanning a broad (60 cm) region of chromosome 3q. Linkage to chromosome 19 was supported by two phenotypes, femur neck axis length (LOD = 2.8) and femur head width (LOD = 2.8). This study is the first genome screen for loci underlying variation in femoral structure and represents an important step toward identifying genes contributing to the risk of osteoporotic hip fracture in the general population. PMID- 11393796 TI - Bone strength and related traits in HcB/Dem recombinant congenic mice. AB - Fracture susceptibility depends jointly on bone mineral content (BMC), gross bone anatomy, and bone microarchitecture and quality. Overall, it has been estimated that 50-70% of bone strength is determined genetically. Because of the difficulty of performing studies of the genetics of bone strength in humans, we have used the HcB/Dem series of recombinant congenic (RC) mice to investigate this phenotype. We performed a comprehensive phenotypic analysis of the HcB/Dem strains including morphological analysis of long bones, measurement of ash percentage, and biomechanical testing. Body mass, ash percentage, and moment of inertia each correlated moderately but imperfectly with biomechanical performance. Several chromosome regions, on chromosomes 1, 2, 8, 10, 11, and 12, show sufficient evidence of linkage to warrant closer examination in further crosses. These studies support the view that mineral content, diaphyseal diameter, and additional nonmineral material properties contributing to overall bone strength are controlled by distinct sets of genes. Moreover, the mapping data are consistent with the existence of pleiotropic loci for bone strength related phenotypes. These findings show the importance of factors other than mineral content in determining skeletal performance and that these factors can be dissected genetically. PMID- 11393797 TI - Inhibition of calcium-stimulated ATPase in the hen brain P2 synaptosomal fraction by organophosphorus esters: relevance to delayed neuropathy. AB - Organophosphorus (OP) compounds have been reported to inhibit Ca/Mg-ATPase, but the relevance of this inhibition to organophosphate-induced delayed neuropathy (OPIDN) has not been explored. To determine if inhibition of this enzyme was related to the development of OPIDN, neuropathic and nonneuropathic OP compounds were sted for their ability to inhibit Ca-stimulated ATPase activity in the P2 synaptosomal fraction from hen brain. Following in vitro exposure to 10(-3) to 10(-5) M OP compounds, Ca-stimulated ATPase activity was inhibited by chlorpyrifos, chlorpyrifos-oxon, phenyl saligenin phosphate (PSP), and tri-o tolyl phosphate (TOTP), but not by parathion, paraoxon, or diisopropyl fluorophosphate (DFP). Further investigation of inhibition induced by chlorpyrifos determined that inhibition was noncompetitive with respect to calcium and ATP. OP compound hydrophobicity was well correlated with in vitro inhibition of Ca-stimulated ATPase, suggesting that OP compounds interact with membrane lipids, and this interaction may contribute to the noncompetitive inhibition of Ca-stimulated ATPase that was observed. Subsequent to in vivo exposure, DFP, but not PSP, produced inhibition of Ca-stimulated ATPase activity in the hen brain P2 synaptosomal fraction. These data indicate that inhibition of Ca-stimulated ATPase activity is not correlated to neuropathic potential and demonstrate that inhibition of Ca/Mg-ATPase is not responsible for OPIDN. PMID- 11393798 TI - Diesel exhaust (DE) affects the regulation of testicular function in male Fischer 344 rats. AB - To investigate the effects of diesel exhaust (DE) particles on the reproductive system, male Fischer 344 rats at 13 mo of age were exposed to clean air or DE at particle concentrations of 0.3, 1, or 3 mg/m3 for 8 mo. DE did not markedly affect testicular and body weights. However, DE at 0.3 mg/m3 significantly decreased prostate and coagulating gland weights, accompanied by a reduction in thymus and adrenal gland weight. In contrast, there was a significant rise in the weights of prostate, seminal vesicles, and coagulating glands in the 3 mg/m3 DE group. In rats exposed to 0.3 or 1 mg/m3 DE, serum luteinizing hormone (LH) and testosterone increased significantly, while a rise in testicular testosterone was noted with 3 mg/m3 DE. The concentrations of follicle-stimulating hormone (FSH) and inhibin as well as the sperm head counts were not markedly altered in any treatment group. Positive staining with inhibin-alpha subunit and 3beta hydroxysteroid dehydrogenase (3beta-HSD) were observed in Sertoli cells and Leydig cells, respectively. Immunolocalization of inhibin-alpha subunit and 3beta HSD was not changed by exposure to DE. In conclusion, DE appears to exert greater effects on accessory glands than on testes in Fischer 344 rats, and the responsiveness of rats is less than that found in mice. PMID- 11393799 TI - Two-generation reproductive toxicity study of tributyltin chloride in female rats. AB - A two-generation reproductive toxicity study of the effects of tributyltin chloride (TBTCl) was conducted in female rats using dietary concentrations of 5, 25, and 125 ppm TBTCl. Reproductive outcomes of dams (number and body weight of pups and the percentage of live pups) and the growth of female pups (the day of eye opening and body weight gain) were significantly decreased in the 125 ppm TBTCl group. A delay in vaginal opening and impaired estrous cyclicity were also observed in the 125 ppm TBTCl group. However, an increase in anogenital distance was found in all TBTCl groups on postnatal d 1. A dose-effect relationship was observed in TBTCl-induced changes in anogenital distance. These results indicate that the whole-life exposure to TBTCl affects the sexual development and reproductive function of female rats. In addition, the TBTCl-induced increase in anogenital distance seems to suggest it may exert a masculinizing effect on female neonates. However, the concentrations of TBTCl used in this study are not environmentally relevant. PMID- 11393800 TI - Immunohistochemical localization of trichloroacylated protein adducts in tetrachloroethene-treated mice. AB - Tetrachloroethene (PCE), a common industrial solvent and environmental contaminant, is primarily used in the dry-cleaning industry. The toxicity of PCE has been linked to vision disorders, renal and hepatic cancer, and autoimmune diseases. Although the mechanism of toxicity is not fully understood, PCE forms trichloroacylated protein adducts in tissues where toxicity is known to occur. These adducts may be responsible for toxicity by altering the function of cellular proteins. Using Western blot analysis, formation of trichloroacylated protein adducts has been reported. To determine the localization of the adducts in a specific zone of a tissue, immunohistochemical staining was used in the study. An antiserum to trichloroacylated proteins was raised in rabbits and its specificity was established by enzyme-linked immunosorbent assay (ELISA). Female MRL-lpr/lpr and MRL +/+ mice were treated with PCE using a single 5-mmol/kg dose over 24 h or on every fourth day for 6 wk (total 20 doses). Formation of trichloroacylated protein adducts was observed in the liver, and localized to the centrilobular zones. Intensity and circumference of the staining around the central vein were much greater in subchronically treated mice than in acutely treated mice. No immunochemical reactivity was observed in any of the other tissues examined. This study shows that hepatic trichloroacylated protein adducts are localized in a region of the liver where PCE-mediated toxicity is known to occur. Immunohistochemical localization of these adducts and its association with PCE-induced toxicity support the contention that adducts may contribute to toxicity. PMID- 11393802 TI - Unusual leg malformations in screech owls from a South Carolina Superfund site. AB - In 1995, the discovery of leg malformations in several screech owl (Otis asio) nestlings and in their female parent at a Department of Energy (DOE) Superfund site in South Carolina prompted an investigation into the nature of the observed abnormalities. Surviving nestlings and the female parent were transferred to a captive screech owl breeding colony at the USGS Patuxent Wildlife Research Center, Laurel, MD. The malformed female parent and her offspring were each mated with normal owls from the colony for 3 yr. Matings of the malfored female produced five malformed and six normal owls; all owls produced by matings of normal offspring were normal. Malformed offspring were euthanized when it became apparent that their physical distress precluded survival under normal conditions of colony care. Euthanized owls were necropsied and examined for skeletal development. Detailed descriptions of eight malformed owls are presented. Results of the matings indicated that the leg mafformations were caused by a genetic trait in the female parent that was heterozygous dominant. The characteristic was lethal except in occasional mild manifestations and resembled an extreme form of a dominant abnormality previously described for domestic fowl called duplicate polydactyly. Other reports of skeletal abnormalities in wild birds and potential environmental causes of genetic mutations at the DOE Superfund site are presented. Other studies performed at the DOE Superfund site do not implicate elevated (above background) ionizing radiation from 137Cs, the dominant radionuclide where the owls were captured, as the cause of the mutation. The cause of this genetic abnormality remains unknown. PMID- 11393801 TI - Evaluation of benzene exposure in children living in Campania (Italy) by urinary trans,trans-muconic acid assay. AB - The urinary benzene metabolite trans,trans-muconic acid (MA) was determined in 144 children living in Campania (Italy): 92 from Naples (1,300,000 inhabitants), designated as an urban source, and compared to 52 from Pollica (300 inhabitants), considered a rural, background exposure for benzene. The children participating in the study were tested by an anonymous questionnaire about the possible sources of exposure to benzene. Quantifiable levels of MA were found in 63% of the urine samples analyzed. Setting the value of nondetectable urinary samples at 7 microg/L MA, a value that is one-half of the instrument detection limit of 14 microg/L, the mean urinary concentration levels were 98.7+/-81.0 microg/L and 48.4+/-71.7 microg/L in Naples and Pollica, respectively; adjustment of these values to creatinine clearance resulted in MA levels of 141.2+/-145.4 microg/L in Naples and 109.8+/-133.2 microg/L in Pollica. Passive smoke exposure did not significantly affect urinary MA levels, but proximity of the home to traffic increased urine MA content. Data show that MA can be utilized as a biomarker for exposure; however, a clear-cut association to benzene requires personal monitoring and control of dietary sorbic acid. PMID- 11393803 TI - Stem cells and the promise of eternal youth: embryonic versus adult stem cells. PMID- 11393804 TI - Hematopoietic stem cells. AB - The adequate production of blood cells is maintained by a set of immature hematopoietic stem cells (HSC) located in the bone marrow after birth. HSC are able to reconstitute the hematopoietic system in disease-related bone marrow failure and bone marrow aplasia. Nowadays, HSC cells can be mobilized from the bone marrow into the peripheral blood using hematopoietic cytokines, allowing a convenient harvest of these cells for clinical transplantation. This review outlines the development of the hematopoietic system in the embryo and in adults and the characterization, enumeration, purification and ex vivo expansion of HSC for clinical use. Future directions include the genetic manipulation of HSC and the identification/expansion of bone marrow-derived stem cells capable of generating non-hematopoietic tissues. PMID- 11393805 TI - Interest of cord blood stem cells. AB - Interest in cord blood stem cells was raised because of the possibility, now realised, of their use in clinical transplantation. The availability of only limited numbers of stem cells in cord blood compared to bone marrow or peripheral blood apheresis after cell mobilisation, led to experimental approaches that first aimed to characterise and then manipulate the stem cells present in cord blood. Their phenotypical and functional characteristics are not identical to those of stem cells in the bone marrow or those cells mobilised into the circulation. The cells selected for phenotype plus Go status show the higher capacity to generate progenitor cells in vitro and will offer the opportunity for mechanistic studies of stem cell self-renewal and proliferation. Another important field of exploration is to investigate the capacity of stem cells in cord blood for differentiation to tissues other than haemopoietic and to establish whether haemopoietic and non-haemopoietic lineages originate in truly multipotential cells or in cells coexisting in cord blood, which have already been limited to differentiation into specific tissue. PMID- 11393806 TI - The neural stem cells and their transdifferentiation capacity. AB - Stem cells play a critical role during embryo and tissue formation throughout development. Thanks to their multipotentiality - i.e., the ability to give rise to different lineages of mature cells - and to their extensive capacity for self renewal and expansive growth, stem cells can also contribute to the maintenance of tissue integrity in adulthood. Historically, it has been held that fetal and adult (somatic) stem cells are tissue-specific 'entities' whose differentiation potential is limited to the generation of mature cell types of the tissue/organ in which they reside. Yet, recent years have seen the publication of an impressive sequence of reports dealing with what is now emerging as one of the most striking functional attributes of somatic stem cells, that is, their capacity to undergo transdifferentiation. Thanks to this peculiar characteristic adult stem cells display an unexpected ability to give rise to differentiated cells of tissues and organs different from those in which they reside. This commentary briefly illustrates the characteristics of the neural stem cell and its capacity as a neuroectodermal derivative to undergo transdifferentiation, thus giving rise to differentiated cells that normally originate from the mesoderm, like blood or skeletal muscle cells. PMID- 11393807 TI - Stem cells and diabetes. AB - Diabetes mellitus is a metabolic disorder affecting 2-5% of the population. Transplantation of isolated islets of Langerhans from donor pancreata could be a cure for diabetes; however, such an approach is limited by the scarcity of the transplantation material and the long-term side effects of immunosuppressive therapy. These problems may be overcome by using a renewable source of cells, such as islet cells derived from stem cells. Stem cells are defined as clonogenic cells capable of both self-renewal and multilineage differentiation. This mean that these cells can be expanded in vivo or in vitro and differentiated to produce the desired cell type. There exist several sources of stem cells that have been demonstrated to give rise to pluripotent cell lines: 1) embryonic stem cells; 2) embryonic germ cells; 3) embryonic carcinoma cells; and 4) adult stem cells. By using in vitro differentiation and selection protocols, embryonic stem cells can be guided into specific cell lineages and selected by applying genetic selection when a marker gene is expressed. Recently, differentiation and cell selection protocols have been used to generate embryonic stem cell-derived insulin-secreting cells that normalise blood glucose when transplanted into diabetic animals. Some recent reports suggest that functional plasticity of adult stem cells may be greater than expected. The use of adult stem cells will circumvent the ethical dilemma surrounding embryonic stem cells and will allow autotransplantation. These investigations have increased the expectations that cell therapy could be one of the solutions to diabetes. PMID- 11393808 TI - Effect of L-lysine and L-arginine on primary osteoblast cultures from normal and osteopenic rats. AB - A therapeutic role of amino acids L-lysine (Lys) and L-arginine (Arg) in osteoporosis and fracture healing was demonstrated previously by in vivo studies. In the present study, primary cultures of osteoblasts were used to investigate the effect of amino acids on gene expression (alkaline phosphatase activity, ALP; osteocalcin, OC; type I collagen), nitric oxide production (NO) and proliferation (MTT) of cells. Cells were isolated from the distal femurs of normal and osteopenic rats. Normal and osteopenic bone-derived cells were divided into four groups: control, Lys (0.587 mg/mL/d), Arg (0.625 mg/mL/d), and Lys + Arg (0.587 + 0.625 mg/mL/d). No evidence of differences between normal and osteopenic bone derived cultures in basal conditions was observed. A significant (P = 0.002) increase of 10.4% in NO production was observed in normal bone-derived osteoblasts treated with Lys + Arg when compared to the control group at 7 days. At the same time, normal bone-derived osteoblasts treated with Arg and Lys + Arg showed significant increases in type I collagen synthesis of 25.3% and 28.4%, respectively, when compared to the control group. Osteopenic bone-derived osteoblasts showed significant (P = 0.002) increases of 27.6% in MTT and 28.7% in cell count at 48 hours when treated with Lys + Arg in comparison with the control group. At 7 days, NO production and type I collagen synthesis increased significantly (P< 0.005) both in osteopenic bone-derived osteoblasts treated with Arg (NO: 18.5%; type I collagen: 34.4%) and Lys + Arg (NO: 23.7%; type I collagen: 20.9%) compared to the control group. Finally, a significant (P = 0.025) decrease of 5.8% in OC level was observed in osteopenic bone-derived osteoblasts treated with Arg. Results suggest that the potential therapeutic effect of Lys and Arg on bone could be related, at least in part, to an improvement of NO production and type I collagen synthesis by osteoblasts both in normal and in osteopenic bone. In osteopenic bone-derived osteoblasts this synthetic phase is preceded by an initial increase of cell proliferation. PMID- 11393809 TI - [3H]Flunitrazepam binding to recombinant alpha1beta2gamma2S GABAA receptors stably expressed in HEK 293 cells. AB - The interaction of selected compounds with the binding of the benzodiazepine [3H]flunitrazepam to membranes isolated from human embryonic kidney (HEK) 293 cells, stably transfected with the aI( 2 2S subtype of GABAA receptors, was studied. This subtype of GABAA receptors is the most common type of GABAA receptor found in the brain, and benzodiazepines are drugs known to enhance the effects of the inhibitory neurotransmitter gamma-amino butyric acid (GABA) by binding to the benzodiazepine binding sites which are part of the GABAA receptor complex. Scatchard analysis of binding data revealed the existence of a single type of binding site for [3H]flunitrazepam. GABA and thiopental enhanced, while the antagonist of central benzodiazepine binding sites--flumazenil, benzodiazepines such as clonazepam, flunitrazepam and diazepam, and the triazolopyridazine CI 218,872--displaced with nanomolar potency the binding of [3H]flunitrazepam. A partial displacement was obtained with the antagonist of the peripheral benzodiazepine binding sites--PK 11195--and with the neurosteroid dehydroepiandrosterone sulfate. The potency of drugs to enhance or inhibit [3H]flunitrazepam binding mainly corresponded to that observed for the modulation of the binding of [3H]flunitrazepam to the native type 1 benzodiazepine binding sites. This, as well as a high density of expressed binding sites, makes the cell line under study a very reliable and economical model for the testing of effects of different compounds at the GABAA receptor. PMID- 11393810 TI - Occult metastasis. AB - The most important factor affecting the outcome of patients with invasive cancers is whether the tumor has spread, either regionally (to regional lymph nodes) or systemically. However, a proportion of patients with no evidence of systemic dissemination will develop recurrent disease after primary 'curative' therapy. Clearly, these patients had occult systemic spread of disease that was undetectable by methods routinely employed (careful pathological, clinical, biochemical and radiological evaluation). In addition, the success of adjuvant therapy is assumed to stem from its ability to eradicate occult metastases before they become clinically evident [1]. Therefore, methods for the detection of occult metastases in patients with the earliest stage of cancer, i.e., prior to detection of metastases by any other clinical or pathological analysis, have received a great deal of attention. PMID- 11393811 TI - The use of Ag85 complex as antigen in ELISA for the diagnosis of bovine tuberculosis in dairy cows in Brazil. AB - Several enzyme-linked immunosorbent assay (ELISA) methods have been investigated to evaluate their performance in the diagnosis of tuberculosis in cattle. Increased production of antibodies to the proteins of antigen 85 complex (Ag85) after experimental and natural infection in cattle shows that they are strongly immunogenic in vivo. The purpose of this study was to evaluate the use of Ag85 as an antigen in ELISA for the diagnosis of bovine tuberculosis in dairy cows in Brazil. The test groups consisted of 46 serum samples from intradermal tuberculin test (ITT)-positive animals (Group A) and 46 samples from ITT-negative animals (Group B). Group C comprised 12 samples from a tuberculosis-free herd and was the control group of the study. Samples were tested in an ELISA using Ag85 as antigen. Differences between the mean ODs of groups A and B and A and C were significant (P < 0.01), but no significant difference (P > 0.05) was observed between groups B and C. The sensitivity of the ELISA using Ag85 was 91.3% (42/46) and its specificity was 94.8% (55/58). These results were not significantly different (P > 0.05) from those observed in a previous study of an ELISA using purified protein derivative (PPD). We concluded that, although Ag85 can be used as antigen for ELISA tests in the diagnosis of bovine tuberculosis with good sensitivity and specificity rates, no significant advantages were observed in relation to the ELISA using PPD that could justify the purification and utilization of Ag85 as a single antigen in routine methods of diagnosis. PMID- 11393812 TI - Pretreatment of guinea-pigs with iron or desferal influences the course of Yersinia enterocolitica infections. AB - The effects of iron excess and desferrioxamine in pretreated guinea-pigs on the immune response (production of Yops) and on the histological changes in infections with Yersinia enterocolitica 0:3 and Y. enterocolitica 0:8 were investigated. The prior overload of the guinea pigs with Dextrofer or treatment with Desferal increased the pathogenic activity of Y. entercolitica 0:3 and led to a generalized infection. Immunoblot analysis showed that in conditions of iron overload the expression of outer membrane proteins (Yops) of Y. enterocolitica 0:8 was blocked. This was accompanied by weak changes in the tissues. The iron limited conditions stimulated production of a low molecular weight protein (17 kDa) on day 6 and easier proliferation of the bacterium. This in vivo study intends to show that in Y. enterocolitica infections a leading role is played not only by iron itself but also by the bacterial strain. PMID- 11393813 TI - Immunogenicity and efficacy of the oral rabies vaccine SAD B19 in foxes. AB - Studies on the immunogenicity and efficacy of SAD B19 attenuated rabies virus vaccine in foxes under laboratory conditions were conducted. Twenty-seven foxes (Vulpes vulpes) were offered a vaccine bait containing 10(6.3) FFU/ml SAD B19. Blood samples were collected 60, 110 and 190 days post-vaccination. On day 190 post vaccination the animals and 14 controls were challenged with a canid street rabies virus. Twenty-four of the 26 vaccinated foxes (92.3%) survived the challenge, whereas all the controls died from rabies. The two vaccinated foxes that did not survive the challenge did not show any detectable level of rabies neutralizing antibodies at any time after vaccination. The geometric mean titres (GMT) of foxes that seroconverted after vaccination were 43.5, 33.9 and 43.5 IU/ml 60, 110 and 190 days post-vaccination, respectively. Furthermore, to test the vaccine virus under sub-optimal conditions five naive and nine previously vaccinated vixens received 2 ml SAD B19 (10(6.7) FFU/ml) by direct administration of the vaccine virus into the oral cavity shortly before or during pregnancy. All vixens seroconverted above the threshold of 0.5 IU/ml. No booster effect was observed in the immune response of the previously vaccinated animals. PMID- 11393814 TI - Immune reactions in cattle after immunization with a Mycobacterium paratuberculosis vaccine and implications for the diagnosis of M. paratuberculosis and M. bovis infections. AB - After immunization of four calves with a live modified Mycobacterium paratuberculosis vaccine the course of the humoral and cell mediated immune reactions was studied during a 2-year clinical investigation. Furthermore, the possibility of shedding of the vaccine strain and the influence of the vaccination on the tuberculin skin test was determined. In addition to standard procedures recently developed diagnostic methods (antibody enzyme-linked immunosorbent assay, interferon-gamma test, polymerase chain reaction) were used. A cell-mediated immune reaction, reflected in an increased, specifically induced, interferon-gamma production developed much earlier (1-2 weeks post-immunization) than humoral immunity (8-16 weeks post-gamma immunization). While the increase in antibody titres was transient, declining to extremely low levels 48-60 weeks post immunization, cell-mediated immunity remained detectable until the end of the investigation. Spread of the vaccine strain into the body and shedding were never detected during the whole course of the study except for one colon site in one calf. As late as 2 years after vaccine application positive or doubtful skin reactions against M. bovis purified protein derivative were measured, reflecting possible interference of the immunization with the diagnosis of bovine tuberculosis. At the end of the investigation, a positive cell-mediated immune reaction was detected the control animal although clinical, pathological and bacteriological examinations gave no indication for a mycobacterial infection. PMID- 11393815 TI - Species of flea (siphonaptera) infesting pets and hedgehogs in Germany. AB - The species of flea infesting pets and hedgehogs in Germany were investigated through a survey of small animal practitioners throughout the country who were asked to collect specimens at their veterinary practices. A total of 625 veterinarians/veterinary practices responded and provided 2445 intact anti identifiable flea specimens. These fleas originated from 294 dogs (795 fleas), 334 cats (1152 fleas), 76 hedgehogs (481 fleas), five domestic rabbits (10 fleas), one golden hamster (four fleas) and one ferret (three fleas). Dogs were found to be infested with Archaeopsylla erinacei, Chaetopsylla globiceps, Ctenocephalides canis, Ctenocephalides felis, Hystrichopsylla talpae, Nosopsyllus fasciatus, Paraceras melis and Pulex irritans. From cats, Archaeopsylla erinacei, Ceratophyllus gallinae, Ceratophyllus garei, Ctenocephalides felis, Ctenophthalmus assimilis, Hystrichopsylla talpae, Monopsyllus sciurorum, Nosopsyllus fasciatus, Spilopsyllus cuniculi and Typhloceras poppei were collected. In both dogs and cats the most prevalent species were Ctenocephalides felis (78.9% and 91.6%, respectively) and Archaeopsylla erinacei (21.1% and 12.6%, respectively) followed by Ctenocephalides canis in dogs (5.8%) and Hystrichopsylla talpae in cats (1.2%). The fleas isolated from rabbits were Ctenocephalides felis, Hystrichopsylla talpae and Spilopsyllus cuniculi. Nosopsyllus fasciatus and Ctenocephalides felis were recovered from the golden hamster and the ferret, respectively. The hedgehogs were found to be infested with Archaeopsylla erinacei, Ceratophyllus gallinae and Ctenocephalides felis. PMID- 11393816 TI - Isolation of pathogenic yersiniae from wild animals in Bulgaria. AB - Pathogenic Yersinia strains were isolated between December 1998 and April 1999 from 37 wild animals: rabbit (Lepus europeus), boar (Sus scrofa scrofa), asiatic jackal (Canis aureus), red fox (Vulpes vulpes), mouflon (Ovis musimon), european river otter (Lutra lutra), beech marten (Martes foina), polecat (Musleta putorius) and wild cat (Felis silvestris). It was established that among the wild animals Y. enterocolitica strains of serotype 0:3 predominated, accompanied by Y. pseudotuberculosis strains of serotype 0:3. In one sample from asiatic jackal and one sample from rabbit, Y. enterocolitica serotype 0:8 was isolated. Yersinia enterocolitica and Y. pseudotuberculosis strains were isolated from tonsils and tongues as well as from the viscera--lung, liver, heart, spleen, kidney and lymph nodes, mainly in young animals (1-2 years of age). The results showed that wild animals are a possible natural reservoir for pathogenic Y. enterocolitica and Y. pseudotuberculosis and are included in the epidemiological chain of yersinioses. PMID- 11393817 TI - Molecular and antigenic characterization of Bangladeshi isolates of infectious bursal disease virus demonstrate their similarities with recent European, Asian and African very virulent strains. AB - Three isolates of infectious bursal disease virus (IBDV), obtained from chickens in Bangladesh in 1999 and designated as BD 1/99, BD 2/99 and BD 3/99, were characterized. In an antigen capture enzyme-linked immunosorbent assay using a panel of VP2-specific neutralizing monoclonal antibodies (mAb), all three isolates showed a mAb-binding profile similar to that of very virulent IBDV (vvIBDV) strains. In contrast to the classical virulent strains, they did not react with mAb 3 and mAb 4. Molecular characterization was performed by direct sequencing of a 677-base pair cDNA corresponding to the VP2 variable domain of the polyprotein gene, synthesized by a reverse transcription-polymerase chain reaction. In comparison to the classical virulent strains, the Bangladeshi isolates were found to have five amino acid substitutions in this region. Four of these changes, Pro222Ala, Val256Ile, Leu294Ile and Asn299Ser, were also observed in other vvIBDV strains. The fifth substitution, Glu300Ala, was similar to that in some African strains of IBDV. The results support the observation that antigenically and genetically similar vvIBDV strains, first observed in Europe in the late 1980s, have spread to most parts of the world in a short period of time. PMID- 11393818 TI - Prevalence of sarcocysts in one-humped camel (Camelus dromedarius) from southern Ethiopia. AB - A survey of sarcocystis was made in camels from southern Ethiopia during a part of 1998-99. A total of 605 haematoxylin and eosin-stained tissue samples from cardiac, diaphragm, shoulder, masseter and oesophagus muscles of 121 adult camels and 20 tissue samples from four foetuses were examined for sarcocysts. Sarcocysts were detected in 55 of 121 (45.45%) camels examined. The infestation rate of oesophagus, diaphragm, shoulder, masseter and cardiac musculatures were 19.83, 11.57, 12.4, 8.26 and 9.17%, respectively. There was no significant (P > 0.05) variation between males (48.6%) and females (40.82%), nor between the two sites studied (Dollo Addo, 40.00% versus Neghelle Borana 47.25%). None of the 20 tissue samples from the four foetuses examined harboured sarcocysts. The possible impact of sarcocysts on camel production is indicated. This is the first report of the presence of sarcocysts in camels from Ethiopia. PMID- 11393819 TI - Bifidobacteria are obligate inhabitants of the crop of adult laying hens. AB - Modified TPY agar (MTPY; with addition of glacial acetic acid and mupirocin) was used in the enumeration of bifidobacteria from hen crop and faeces. The colonies on MTPY medium inoculated with crop and faeces samples were Gram-stained, screened for presence of fructose-6-phosphate phosphoketolase activity and tested for fermentation patterns using ANAEROtest kits. It was revealed that bifidobacteria are obligate inhabitants of the hen crop, reaching counts of 10(7)/g of crop content. The occurrence of bifidobacteria in the hen crop was evidently not the consequence of ingestion of faeces as all the bifidobacteria strains isolated from the hen crop fermented glucose and fructose, while most strains isolated from hen faeces did not ferment either glucose or fructose. The results suggest that bifidobacteria are common and may even be one of the predominant parts of the hen crop flora. PMID- 11393820 TI - Capillaria hepatica infection in wild brown rats (Rattus norvegicus) from the urban area of Milan, Italy. AB - Forty-seven wild brown rats (Rattus norvegicus) collected from the urban area of Milan (Italy) were screened for Capillaria hepatica liver infection. The liver of each rat was grossly and histologically examined for the presence of C. hepatica adults, eggs and typical C. hepatica induced lesions. In 17 rats (36%) liver lesions consistent with C. hepatica infection were detected. Grossly, white yellow nodules of 1-5 mm in diameter were present, either scattered on the liver surface or localized in a single lobe. Histologically, granulomatous liver lesions associated with eggs and/or worms were observed. The degree of gross liver involvement was moderate in most of the positive cases (71%). About 30 cases of C. hepatica infection in humans have been documented world-wide, most of which are reported in children from 1 to 5 years of age. Our results suggest that the potential transmission of C. hepatica to children in the study area should be considered an important health issue. PMID- 11393821 TI - The influence of host haematocrit on the blood feeding success of Anopheles stephensi: implications for enhanced malaria transmission. AB - Two studies were carried out to determine the effect of the rodent malaria Plasmodium yoelii nigeriensis on the blood feeding success of Anopheles stephensi. Initially, pairs of mice with similar packed cell volume (PCV) (measured by haematocrit) were selected. Following infection of one of the pair its PCV gradually fell. At various times post-infection, a comparison was made of the bloodmeal size (haemoglobin content) of mosquitoes feeding on these mice. The bloodmeal sizes increased with parasite-induced fall in PCV down to a haematocrit of 43-44%, which occurred approximately 48 h post-infection. Bloodmeals were significantly reduced, however, when mosquitoes fed on mice with higher parasitaemias and a haematocrit of 15-35%. Thus, at early stages of infection, mosquitoes ingested a bloodmeal significantly greater than did the mosquitoes feeding on the control mice. However, mosquitoes were not able to compensate for severe infection-associated anaemia. To compensate for variation due to innate differences in the mice, a second experiment was performed. Mosquitoes were fed on the same mice before (control) and after infection. Again, the bloodmeal size increased with decreasing PCV down to haematocrits of 42-45%, but declined thereafter. In this host-parasite-vector system, haematocrits that maximized erythrocyte intake were produced when gametocytes, capable of exflagellation, were present. PMID- 11393822 TI - Mutation rates in the dihydrofolate reductase gene of Plasmodium falciparum. AB - A new method has been established to define the limits on a spontaneous mutation rate for a gene in Plasmodium falciparum. The method combines mathematical modelling and large-scale in vitro culturing and calculates the difference in mutant frequencies at 2 separate time-points. We measured the mutation rate at 2 positions in the dihydrofolate reductase (DHFR) gene of 3D7, a pyrimethamine sensitive line of P. falciparum. This line was re-cloned and an effectively large population was treated with a selective pyrimethamine concentration of 40 nM. We detected point mutations at codon-46 (TTA to TCA) and codon-108 (AGC to AAC), resulting in serine replacing leucine and asparagine replacing serine respectively in the corresponding gene product. The substitutions caused a decrease in pyrimethamine sensitivity. By mathematical modelling we determined that the mutation rate at a given position in DHFR was low and occurred at less than 2.5 x 10(-9) mutations/DHFR gene/replication. This result has important implications for Plasmodium genetic diversity and antimalarial drug therapy by demonstrating that even with low mutation rates anti-malarial resistance will inevitably arise when mutant alleles are selected under drug pressure. PMID- 11393823 TI - Landscape features associated with infection by a malaria parasite (Plasmodium mexicanum) and the importance of multiple scale studies. AB - In a 3-year study, we examined landscape features (aspect, slope, sun exposure, canopy cover, type of ground cover, and nearest water source) that were potentially related to prevalence of infection with Plasmodium mexicanum in fence lizards (Sceloporus occidentalis) within a 4.5 ha study area in northern California, USA. Logistic regression analysis showed that ground cover type was the primary mediator of the probability of P. mexicanum infection. Infected lizards were captured more often in rock and/or leaf litter locations than in grassy ones. In another experiment, the study area was divided into 9 sites (0.07 0.33 ha), and infection prevalence was calculated for each. Three sites with high (> 30%) infection prevalence had significantly more rocky outcrops and leaf litter than those with low (< 20%) or moderate (20-30%) infection prevalence (N = 3 sites each). We conclude that lizard site selection may influence the probability of exposure to infected vectors and thus the likelihood of P. mexicanum infection. We also demonstrate that studies at different spatial scales may be required to understand fully the relationship between landscape features and parasite distribution. PMID- 11393824 TI - Decrease of psychomotor performance in subjects with latent 'asymptomatic' toxoplasmosis. AB - Toxoplasma gondii is known to induce specific behavioural changes in its intermediate hosts. This is usually considered to be an evolutionary adaptation aimed to increase the probability of transmission of the parasite into its definitive host, the cat, by predation. In rodents an increase of reaction time as well as many other specific behavioural patterns have been observed. Here we report the results of our double blind study showing the significantly longer reaction times of 60 subjects with latent toxoplasmosis in comparison with those of 56 controls. Moreover, the existence of a positive correlation between length of infection and mean reaction time suggested that slow and cumulative effects of latent toxoplasmosis rather than a one-step (and possibly transient) effect of acute toxoplasmosis disease are responsible for the decrease of psychomotor performance of infected subjects. To our knowledge, this is the first study confirming the existence of such parasite-induced changes in human behaviour that could be considered in evolutionary history of the human species as adaptive from the point of view of parasite transmission. PMID- 11393825 TI - Transformation of Leishmania mexicana metacyclic promastigotes to amastigote-like forms mediated by binding of human C-reactive protein. AB - Infective metacyclic promastigote forms of Leishmania mexicana are introduced by the bite of sandfly vectors into their human hosts where they transform into the amastigote form. The kinetics of this process was examined in vitro in response to different combinations of temperature (26 degrees C or 32 degrees C), pH (7.2 or 5.5), and exposure to human serum. Little transformation occurred at 26 degrees C/pH 7.2, intermediate levels at 26 degrees C/pH 5.5 and 32 degrees C/pH 7.2, and the greatest response at 32 degrees C/pH 5.5. Transformation was stimulated by exposure to normal human serum, but was markedly reduced when serum previously incubated at 56 degrees C for 1 h was used (complement heat inactivated). This stimulatory effect was reproduced by exposure to a single purified component of human serum, C-reactive protein (CRP). Binding of CRP to the whole surface of L. mexicana metacyclic promastigotes, including the flagella, was demonstrated by an indirect fluorescent antibody test. The effect of purified CRP was dose dependent and occurred using normal serum concentrations. The stimulatory effect of whole serum was oblated by CRP depletion and restored by addition of purified CRP. The effects of cAMP analogues indicated that transformation could be mediated via an adenylate cyclase cascade. PMID- 11393826 TI - The Egyptian mongoose, Herpestes ichneumon, is a possible reservoir host of visceral leishmaniasis in eastern Sudan. AB - Investigations were made on possible reservoir hosts of Leishmania donovani in 2 zoonotic foci of visceral leishmaniasis (VL) in Dinder National Park (DNP) and the peri-domestic habitats of adjacent villages of eastern Sudan. Animals were captured, in November 1997-1998 and April-May 1999 and examined for L. donovani infection using light microscopy and 2 sensitive Polymerase Chain Reaction (PCR) systems. Microscopy and PCR investigations were also used to determine the infection rates of L. donovani in Phlebotomus orientalis captured from the uninhabited site of DNP. Infections of L. donovani were detected in 2 out of 14 Egyptian mongooses (Herpestes ichneumon), 1 out of 168 Arviconthus niloticus and 1 out of 8 Mastomys natalensis. Samples from 68 other animals captured from the study area were all negative for the infection. Active zoonotic transmission of L. donovani at the time of animal sampling in the uninhabited site of DNP was demonstrated by finding the parasite in 3.4% (7 out of 184) and 3.2% (5 out of 157) of flies collected in March 1998 and May 1999, respectively. We suggest that the Egyptian mongoose is a possible reservoir host of L. donovani. The importance of other animals in maintaining the infection is also discussed. PMID- 11393827 TI - Relative contribution of day-to-day and intra-specimen variation in faecal egg counts of Schistosoma mansoni before and after treatment with praziquantel. AB - There is evidence that faecal egg counts of Schistosoma mansoni vary considerably from day to day, which results in poor sensitivity of single stool readings. Intra-specimen variation of S. mansoni egg counts may also be considerable, but has previously been considered as the less important component. We quantified the relative contribution of these two sources of variation among 96 schoolchildren from an area in Cjte d'Ivoire highly endemic for S. mansoni. Stool specimens were collected over 5 consecutive days, and 5 egg-counts were made in each specimen by the Kato-Katz technique. The point prevalence of the first sample was 42.7% and the cumulative prevalence after the maximum sampling effort was 88.5%. Using generalized linear mixed models we found that the presence of S. mansoni eggs in a stool sample varied much more between days than within specimens, indicating that stool sample examination over multiple days is required for accurate prevalence estimates. However, using the same approach, we found that among infected children intra-specimen variation in egg counts was 4.3 times higher than day-to-day variation. After praziquantel administration, day-to-day variation was more important than before, since most infections were very light and thus likely to be missed altogether by stool examination on a single day. We conclude that diagnostic sensitivity in high transmission areas is maximized by making several stool readings on several days, but examining 1 stool specimen several times can make reasonable estimates of infection intensity. PMID- 11393828 TI - Parasite (Schistosoma mansoni) and host (Biomphalaria glabrata) genetic diversity: population structure in a fragmented landscape. AB - Random amplified polymorphic DNA (RAPD) markers were used to quantify genetic diversity within and between 5 populations of Schistosoma mansoni within its definitive host (Rattus rattus) and the 5 corresponding populations of the snail intermediate host (Biomphalaria glabrata) from a limited endemic area of murine schistosomiasis on the island of Guadeloupe. Analysis of molecular variance (AMOVA) and canonical correspondence analysis (CCA) were used to test the significance of genetic differentiation between populations. Both methods gave similar results. Of total gene diversity, 15.1% (AMOVA) and 18.8% (CCA) was partitioned between localities for S. mansoni with an absence of association between genetic and geographical distances. Geographical localities accounted for 20.5% (CCA) of the total diversity for B. glabrata populations. The genetic distances between pairs of parasite populations were not correlated with the genetic distances between the corresponding pairs of snail host populations. Such strong patterns of local differentiation of both parasite and snail populations are consistent with predictions based on metapopulation dynamics and may have implications on host-parasite susceptibility relationship through local adaptation processes. PMID- 11393829 TI - Paramyosin from the parasitic mite Sarcoptes scabiei: cDNA cloning and heterologous expression. AB - The burrowing mite Sarcoptes scabiei is the causative agent of the highly contagious disease sarcoptic mange or scabies. So far, there is no in vitro propagation system for S. scabiei available, and mites used for various purposes must be isolated from infected hosts. Lack of parasite-derived material has limited the possibilities to study several aspects of scabies, including pathogenesis and immunity. It has also hampered the development of high performance serological assays. We have now constructed an S. scabiei cDNA expression library with mRNA purified from mites isolated from red foxes. Immunoscreening of the library enabled us to clone a full-length cDNA coding for a 102.5 kDa protein. Sequence similarity searches identified the protein as a paramyosin. Recombinant S. scabiei paramyosin expressed in Escherichia coli was recognized by sera from dogs and swine infected with S. scabiei. We also designed a small paramyosin construct of about 17 kDa that included the N-terminal part, an evolutionary variable part of the helical core, and the C-terminal part of the molecule. The miniaturized protein was efficiently expressed in E. coli and was recognized by sera from immunized rabbits. These data demonstrate that the cDNA library can assist in the isolation of important S. scabiei antigens and that recombinant proteins can be useful for the study of scabies. PMID- 11393830 TI - Analysis of aggregation, a worked example: numbers of ticks on red grouse chicks. AB - The statistical aggregation of parasites among hosts is often described empirically by the negative binomial (Poisson-gamma) distribution. Alternatively, the Poisson-lognormal model can be used. This has the advantage that it can be fitted as a generalized linear mixed model, thereby quantifying the sources of aggregation in terms of both fixed and random effects. We give a worked example, assigning aggregation in the distribution of sheep ticks Ixodes ricinus on red grouse Lagopus lagopus scoticus chicks to temporal (year), spatial (altitude and location), brood and individual effects. Apparent aggregation among random individuals in random broods fell 8-fold when spatial and temporal effects had been accounted for. PMID- 11393831 TI - A microsatellite polymorphism in the gamma interferon gene is associated with resistance to gastrointestinal nematodes in a naturally-parasitized population of Soay sheep. AB - Free-living Soay sheep (Ovis aries) on the island of Hirta, St Kilda, Scotland, are naturally parasitized by gastrointestinal nematodes, predominantly Teladorsagia circumcincta. In this paper we show that reduced faecal egg counts (FEC) are associated with an allele at a microsatellite locus located in the first intron of the interferon gamma gene (o(IFN)-gamma) in Soay sheep lambs and yearlings, measured at approximately 4 and 16 months of age, respectively. The same allele was also associated with increased T. circumcincta-specific antibody (IgA) in lambs, but not associated significantly in yearlings. Flanking control markers failed to show a significant association with either FEC or IgA. These results suggest that a polymorphic gene conferring increased resistance to gastrointestinal nematode parasites is located at or near the interferon gamma gene, and support previous reports which have mapped a quantitative trait locus (QTL) for resistance to this region in domestic sheep. Our data are consistent with the idea that a functional polymorphism leading to reduced expression or efficacy of (IFN)-gamma could enhance the immune response to gastrointestinal nematodes by favouring the activity of the Th2 cell subset and antibody associated immune mechanisms. PMID- 11393832 TI - Heat shock and stress response of Taenia solium and T. crassiceps (Cestoda). AB - Heat shock and stress responses are documented for the first time in larval stages of the cestodes Taenia solium and Taenia crassiceps. Radioactive metabolic labelling after in vitro incubation of cysts at 43 degrees C, revealed the induction of heat shock proteins. In T. crassiceps, the major heat shock proteins were 80, 70 and 60 kDa. After prolonged incubation, a set of low molecular weight heat shock proteins (27, 31, 33 and 38 kDa), were also induced. In vitro incubation of cysts at 4 degrees C, induced the synthesis of stress proteins ranging from 31 to 80 kDa, indicating the parasite is also able to respond to cold shock. T. solium cysts exposure to temperature stress also resulted in an increased synthesis of 2 major heat shock proteins of 80 and 70 kDa. Western blots using the excretory-secretory products of T. solium showed that 2 heat shock proteins were recognized by antibodies in the sera of cysticercotic patients: one of 66 kDa and another migrating close to the run front. The T. solium 66 kDa protein was also recognized by specific antibodies directed to a 60 kDa bacterial heat shock protein, suggesting that it belongs to this family of proteins. PMID- 11393833 TI - Effect of iron deficiency on Trichuris suis and Ascaris suum infections in pigs. AB - The objective of this experiment was to detect a possible interaction between iron deficiency and intestinal nematode infections. We report on a 2 x 2 study where thirty-one 10-week-old pigs fed a low or a normal iron diet were infected with both Trichuris suis (4500 eggs) and Ascaris suum (1200 eggs). No significant difference was detected between diet groups with respect to parasitological parameters for A. suum or the total number of adult T. suis recovered at necropsy 10 weeks p.i. However, in the low iron group T. suis were located more proximally and the worms were increased in length. A higher proportion of pigs with initial faecal egg excretion at 6 weeks p.i. was observed in the low iron group, indicating a shortened pre-patency period. Worm fecundity and total faecal egg excretion were also highest in the low iron group. A significant correlation was found between female worm length and fecundity. The peripheral eosinophil counts were diminished in the low iron host groups. The infected low iron group experienced more severe pathophysiological changes in terms of hypoalbuminaemia and decreases in erythrocyte volumes. A significant inverse correlation existed between iron content in the bone-marrow and liver (body) store. In conclusion, iron deficiency increased the severity of T. suis infection in pigs. PMID- 11393834 TI - Scanning optical microscopy with an electrogenerated chemiluminescent light source at a nanometer tip. AB - Electrogenerated chemiluminescence at electrodes with effective diameters down to 155 nm was used as a stable light source for near-field scanning optical microscopy imaging of an interdigitated array and a submicrometer size test substrate. Light was generated in a thin (approximately 500 microm) layer of an aqueous solution of 15 mM Ru(bpy)3(2+) and 100 mM tri-n-propylamine in a pH 7.5 buffer. The resolution obtained was compared to that found with a micrometer size electrode. The shear force from the tip attached to a quartz tuning fork was used to monitor and control the tip-to-substrate separation within the near field regime. PMID- 11393835 TI - Analysis of free drug fractions by ultrafast immunoaffinity chromatography. AB - A chromatographic method was developed for measuring the nonbound (or free) fraction of drugs by using millisecond-scale extractions on small immunoaffinity columns. The design of this system was developed by considering the dissociation rates of (R)- and (S)-warfarin from the binding protein human serum albumin (HSA) and by performing computer simulations of the immunoaffinity extraction of these drugs. The final system was tested by using it to measure the free fractions of (R)- or (S)-warfarin in samples with known concentrations of these agents and HSA. The free warfarin fraction was extracted in 180 ms by a 2.1-mm-i.d. sandwich microcolumn that contained a 1.1-mm layer of an anti-warfarin antibody support. The nonretained peaks from this immunoaffinity column were passed through a series internal surface reversed-phase columns and a fluorescence detector for the analysis of any protein-bound warfarin that remained in the sample. The experimental results were found to have good agreement with those predicted from the known equilibrium constants for the binding of (R)- and (S)-warfarin with HSA. This approach can be modified for other analytes by changing the types of antibodies that are used in the immunoaffinity column and by using an appropriate detector for the nonretained drug fraction. PMID- 11393836 TI - A fluorogenic reagent, 4-mercapto-7-methylthio-2,1,3-benzoxadiazole for carboxylic acids, designed by prediction of the fluorescence intensity. AB - During the course of our studies of the development of fluorogenic reagents having a 4,7-disubstituted benzofurazan structure, we previously proposed 7 acetylamino-4-mercapto-2,1,3-benzoxadiazole (AABD-SH) as a fluorogenic reagent for carboxylic acids. Since then, progress has made it possible to estimate the fluorescence quantum yields of the 4,7-disubstituted benzofurazan compounds on the basis of the PM3 calculation of their S1-T2 energies. Subsequently, a new fluorogenic reagent, 4-mercapto-7-methylthio-2,1,3-benzoxadiazole (MTBDSH) was designed and synthesized. In the presence of condensation reagents, triphenylphosphine (TPP) and 2,2'-dipyridyl disulfide (DPDS), MTBD-SH readily reacted with n-caprylic acid within 1 min at room temperature. The derivatives of five carboxylic acids (n-caprylic acid, n-capric acid, lauric acid, myristic acid, and palmitic acid) were well-separated on a reversed-phase column and were fluorimetrically detected at 519 nm with excitation at 391 nm. The detection limits (S/N = 3) were 2.4-5.0 fmol. Thus, MTBD-SH had properties that were considered to be superior. For carboxylic acids, itwas superior not only to AABD SH, but also to many other conventional reagents. The superiority was examined in terms of its reactivity and sensitivity and the avoidance of interfering peaks that were derived from the reagent itself or degradation products in the chromatogram. PMID- 11393837 TI - Determination of clenbuterol in bovine liver by combining matrix solid-phase dispersion and molecular imprinted solid-phase extraction followed by liquid chromatography/electrospray ion trap multiple-stage mass spectrometry. AB - Matrix solid-phase dispersion (MSPD) is a new sample pretreatment for solid samples. This technique greatly simplifies sample pretreatment but, nonetheless, the extracts often still require an extra cleanup step that is both laborious and time-consuming. The potential of combining MSPD with molecularly imprinted solid phase extraction (MISPE) was investigated in this study. Liver samples were ground in a mortar with C18 sorbent and the homogenized mixture packed into an SPE cartridge and placed on top of a MISPE cartridge. Subsequently, clenbuterol was eluted from the MSPD cartridge onto the MISPE cartridge using acetonitrile containing 1% acetic acid. The ability of the molecularly imprinted polymer to selectively adsorb analyte in acetonitrile was exploited for re-extracting clenbuterol directly from this acetonitrile extract via the double cartridge tandem system. The analyte was eluted from the MISPE cartridge using acidified methanol. A clear eluate was obtained, which was subsequently evaporated, redissolved, and analyzed by HPLC electrochemical detection (ECD) or ion trap mass spectrometry (LC/IT-MS). The MISPE cartridge used in this study was imprinted using bromoclenbuterol, a structural analogue of clenbuterol, as the template. These MISPE cartridges showed excellent stability. The complete extraction procedure was rapid, and recoveries exceeded 90% for the target analyte. The method detection limit for the LC/IT-MS procedure was < 0.1 microg/kg. This method, therefore, satisfies the stringent requirements of European Union regulation EEC 2377/90. PMID- 11393838 TI - Simultaneous quantification of etoposide and etoposide phosphate in human plasma by capillary electrophoresis using laser-induced native fluorescence detection. AB - A method using capillary electrophoresis with UV laser-induced native fluorescence detection was developed as a sensitive and selective assay for the simultaneous determination of etoposide and etoposide phosphate in human plasma. Laser-induced native fluorescence detection with a frequency-doubled argon ion laser at an excitation wavelength of 257 nm was used for the simultaneous assay of etoposide and etoposide phosphate in plasma to improve the sensitivity compared to that obtained with UV absorption. The detection system consists of an imaging spectrograph and an intensified CCD camera which views an illuminated 1.5 mm section of the capillary. This setup is able to record the whole emission spectra of the analytes to achieve additional wavelength-resolved electropherograms. In the concentration range of 200 microg/L-50 mg/L in plasma for etoposide and 100 microg/L-20 mg/L for etoposide phosphate, coefficients of correlation were better than 0.998. Within-day variation determined with three different concentrations showed accuracies ranging from 91.0 to 109.3% for etoposide and from 91.2 to 109.9% for etoposide phosphate (n = 6) with a precision of about 8%. Day-to-day variation presented accuracies ranging from 91.8 to 107.9% for etoposide and from 94.4 to 109.3% for etoposide phosphate with a relative standard deviation less than 6% (n = 5). To our knowledge, this is the first method for the simultaneous quantification of etoposide and etoposide phosphate in plasma samples. PMID- 11393839 TI - Miniaturized electrospraying as a technique for the production of microarrays of reproducible micrometer-sized protein spots. AB - Electrospraying in a stable cone-jet mode at <400 microm above a substrate is shown to be a powerful technique to produce arrays of identical micrometer-sized spots consisting of biologically active substances. Aqueous solutions with a surface tension of 0.04 N m(-1) and conductivities ranging from 0.04 to 2.2 S m( 1) were sprayed at ultralow flow rates ranging from 100 to 300 pL s(-1). The charged jet that emanates from the cone tip breaks up into a spray of charged droplets that are deposited in the form of a uniform spot of 130-350 microm in diameter by spraying during 0.5-3 s at 220-400 microm above a substrate, respectively. After a spot was deposited, spraying was stopped instantaneously by increasing the distance between the capillary tip and the substrate by an additional 100 microm using a computer-controlled x-y-z table. This was immediately followed by a rapid shift of the substrate 400 microm sideways and 100 microm upward, thus causing spraying to resume instantaneously because of the increased electric field strength, which resulted in the deposition of the next spot. It is shown here that spraying of lactate dehydrogenase (LDH), glucose-6 phosphate dehydrogenase (G6P-DH), and pyruvate kinase (PK) on a liquid layer resulted in the complete preservation of their activities despite the high solution conductivity of 3.3 S m(-1) and high currents ranging from 300 to 500 nA. LDH and PK activities were fully preserved after spraying onto dry aluminum by adding 0.05 M buffer and 0.5 and 1 wt % of trehalose, respectively, to the spray solutions. Electrospraying allows for accurate dispensing of liquid volumes as small as 50 pL. Enzymatic activities of LDH and PK are fully preserved after spraying. PMID- 11393840 TI - Development of isotope dilution cold vapor inductively coupled plasma mass spectrometry and its application to the certification of mercury in NIST standard reference materials. AB - An isotope dilution cold vapor inductively coupled plasma mass spectrometry (ID CV-ICPMS) method featuring gaseous introduction of mercury via tin chloride reduction has been developed and applied to the quantification and certification of mercury in various NIST standard reference materials: SRM 966 Toxic Metals in Bovine Blood (30 ng x mL(-1)); SRM 1641d Mercury in Water (1.6 microg x mL(-1)); and SRM 1946 Lake Superior Fish Tissue (436 ng x g(-1)). Complementary mercury data were generated for SRMs and NIST quality control standards using cold vapor atomic absorption spectroscopy (CVAAS). Certification results for the determination of mercury in SRM 1641d using two independent methods (ID-CV-ICPMS and CVAAS) showed a degree of agreement of 0.3% between the methods. Gaseous introduction of mercury into the ICPMS resulted in a single isotope sensitivity of 2 x 10(6) counts x s(-1)/ng x g(-1) for 201Hg and significantly reduced the memory and washout effects traditionally encountered in solution nebulization ICPMS. Figures of merit for isotope ratio accuracy and precision were evaluated at dwell times of 10, 20, 40, 80, and 160 ms using SRM 3133 Mercury Spectrometric Solution. The optimum dwell time of 80 ms yielded a measured 201Hg/202Hg isotope ratio within 0.13% of the theoretical natural value and a measurement precision of 0.34%, on the basis of three replicate injections of SRM 3133. PMID- 11393841 TI - Determination of perfluorinated surfactants in surface water samples by two independent analytical techniques: liquid chromatography/tandem mass spectrometry and 19F NMR. AB - Perfluorinated surfactants are an important class of specialty chemicals that have received recent attention as a result of their persistence in the environment. Two analytical methods for the determination of perfluorinated surfactants in aqueous samples were developed in order to investigate a spill of 22000 L of fire retardant foam containing perfluorinated surfactants into Etobicoke Creek (Toronto, Ontario). With the first method, aliquots of surface water (0.2-200 mL) were preconcentrated using solid-phase extraction. Liquid chromatography/tandem mass spectrometry was employed for identification and quantification of each perfluorinated surfactant. Total perfluorinated surfactant concentrations in surface water samples ranged from 0.011 to 2270 microg/L, and perfluorooctanesulfonate was the predominant surfactant observed. Interestingly, perfluorooctanoate was detected in surface water sampled upstream of the spill. A second method employing 19F NMR was developed for the determination of total perfluorinated surfactant concentrations in aqueous samples (2-100 mL). By 19F NMR, the surface water concentrations ranged from nondetect (method detection limit, 10 microg/L for a 100-mL sample) to 17000 microg/L. These methods permit comprehensive evaluation of aqueous samples for the presence of perfluorinated surfactants and have applicability to other sample matrixes. PMID- 11393842 TI - Monitoring DNA immobilization and hybridization on surfaces by atomic force microscopy force measurements. AB - DNA immobilization and hybridization was carried out on Au substrates that were modified with mercaptopropanoic acid and then treated with aluminum(III) solution. The positively charged AI(III) film can be used to immobilize both ds DNA and ss-DNA. Atomic force microscopy (AFM) was used to monitor the process by force measurements between a negatively charged silica tip and the substrates while immersed in dilute electrolyte. Surface hybridization of ss-DNA produces an increase in the surface charge and surface potential of the substrates, which is reflected by the increasing repulsive force as determined from AFM force separation curves. A single-base mismatch was detectable in surface hybridization. The AFM force measuring technique was also employed to investigate the interaction of Ru(phen)3(2+) with ss-DNA and ds-DNA. The force measurement results showed that there is a small interaction between Ru(phen)3(2+) and ss DNA, which was ascribed to the electrostatic binding of Ru(phen)3(2+) to the ss DNA surface. For ds-DNA, there is a strong interaction which is believed to be due to the association or intercalation of Ru(phen)3(2+) with ds-DNA. PMID- 11393843 TI - Chemical image fusion. The synergy of FT-NIR and Raman mapping microscopy to enable a more complete visualization of pharmaceutical formulations. AB - The pharmaceutical industry uses successfully both FT-NIR and Raman microscopy to produce chemical images of solid dosage forms, typically in troubleshooting roles. However, due to the chemical composition of the formulations, it is not always possible to describe the entire chemical formulation by using a single spectroscopic method. As Raman and NIR spectroscopies are complementary in nature, their combined usage offers the opportunity to describe heterogeneous mixtures in more detail. A novel sample referencing approach has been developed that allows data to be acquired from exactly the same area of the sample using both Raman and FT-NIR microscopies. The optimum images for the components are then overlaid, which gives rise to a combined chemical image that visually describes the entire formulation. We have named this approach chemical image fusion (CIF). CIF has been applied to two examples. The first shows how a simple formulation was used to validate the CIF approach. In the second, CIF allowed an entire formulation to be visualized and the cause of tabletting problems determined. CIF provides increased confidence in the results generated by each individual technique and offers a more powerful method for the evaluation of pharmaceutical formulations. PMID- 11393844 TI - Fluorescent nanosensors for intracellular chemical analysis: decyl methacrylate liquid polymer matrix and ion-exchange-based potassium PEBBLE sensors with real time application to viable rat C6 glioma cells. AB - Fluorescent spherical nanosensors, or PEBBLEs (probes encapsulated by biologically localized embedding), in the 500 nm-1 microm size range have been developed using decyl methacrylate as a matrix. A general scheme for the polymerization and introduction of sensing components creates a matrix that allows for the utilization of the highly selective ionophores used in poly(vinyl chloride) and decyl methacrylate ion-selective electrodes. We have applied these optically silent ionophores to fluorescence-based sensing by using ion-exchange and highly selective pH chromoionophores. This allows the tailoring of selective submicrometer sensors for use in intracellular measurements of important analytes for which selective enough fluorescent probes do not exist. The protocol for sensor development has been worked out for potassium sensing. It is based on the BME-44 ionophore (2-dodecyl-2-methyl-1,3-propanediylbis[N-[5'nitro(benzo-15-crown 5)-4'-yl]carbamate]). The general scheme should work for any available ionophore used in PVC or decyl methacrylate ion-selective electrodes, with minor adjustments to account for differences in ionophore charge and analyte binding constant. The reversible and highly selective sensors developed have a subsecond response time and an adjustable dynamic range. Applications to live C6 glioma cells demonstrate their utility; the intracellular potassium activity is followed in real time upon extracellular administration of kainic acid. PMID- 11393845 TI - Detection of abasic sites on individual DNA molecules using atomic force microscopy. AB - We have developed an atomic force microscopy-based method for detecting abasic sites (AP sites) on individual DNA molecules. By using uracil and uracil DNA glycosylase, we first prepared a 250-bp DNA template consisting of two AP sites at specific locations. We then detected the AP sites by marking them with biotinylated aldehyde-reactive probes and monomeric avidin. We demonstrate here that (i) the location of monomeric avidin bound on a single DNA molecule was detectable by atomic force microscopy; (ii) the observed location of avidin was in good agreement to the predicted AP sites at a few nanometer resolution; and (iii) by end-labeling the 5'-terminus of one DNA strand, the AP sites were determined without directional ambiguity. The technique described here will provide a sensitive way of locating AP sites and contribute to screen DNA damages from individual molecules. PMID- 11393846 TI - Surface-induced dissociation on a MALDI-ion mobility-orthogonal time-of-flight mass spectrometer: sequencing peptides from an "in-solution" protein digest. AB - Peptide sequencing by surface-induced dissociation (SID) on a MALDI-ion mobility orthogonal TOF mass spectrometer is demonstrated. SID of approximately 100-fmol amounts of model peptides HLGLAR (m/z 666.8), gramicidin S (m/z 1142.5), and bovine insulin b chain (m/z 3495.5) was accomplished using hydrocarbon-coated gold grids and approximately 20-eV collision energies. The current version of the instrument achieves a mobility resolution of approximately 20 and TOF mass resolution better than 200. Peptide sequences of four peptides from a tryptic digest of cytochrome c (approximately 1 pmol deposited) were obtained. The advantage of IM-SID-o-TOF-MS is that a single experiment can be used to simultaneously measure the molecular weights of the tryptic peptide fragments (e.g., peptide mass mapping) and partial sequence analysis, (e.g., real-time tandem mass spectrometry.) PMID- 11393847 TI - A method for chemometric classification of unknown vapors from the responses of an array of volume-transducing sensors. AB - A method for the characterization and classification of unknown vapors based on the responses on an array of polymer-based volume-transducing vapor sensors is presented. Unlike conventional pattern recognition methods, the sensor array pattern vector is converted into another vector containing vapor descriptors. Equations are developed to show how this approach can be applied to arrays of sensors where each sensor responds to the fractional volume increase of the polymer upon vapor sorption. The vapor sorption step of the response is modeled with linear solvation energy relationships using solvation parameters as vapor descriptors. The response model also includes the vapor concentration, the sensitivity to fractional volume increases, and the specific volume of the vapor as a liquid. The response model can be solved for the vapor descriptors given the array responses and sensitivity factors, following an approach described previously for purely gravimetric sensors. The vapors can then be classified from a database of candidate vapor descriptors. Chemiresistor vapor sensors coated with composite polymer films containing conducting particles represent a volume transducing sensor technology to which this new classification method should apply. Preliminary equations are also presented for sensors that respond on the basis of both the mass and the volume of a sorbed vapor. Surface acoustic wave sensors with acoustically thin polymer films that respond to both mass and modulus effects may fit this classification approach. PMID- 11393848 TI - Analysis of dark spots growing in organic EL devices by time-of-flight secondary ion mass spectrometry. AB - Chemical structural analysis of tape-stripped surfaces at dark spots growing in organic electroluminescent (EL) devices during exposure to the atmosphere was done by time-of-flight secondary ion mass spectrometry (OF-SIMS). The EL devices consist of indium-tin-oxide, triphenylamine-tetramer, tris(8 hydroxyquinoline)aluminum (Alq3), and a Mg-Ag cathode deposited in order under vacuum on a glass substrate. It was found that the interface between the Alq3 layer and the Mg-Ag cathode was exposed as a result of tape-stripping, where a large number of dark spots were observed on both sides. Secondary ion images of O , Mg+, and Alq2+ were observed from the dark spots on the cathode side. On the other hand, Mg+ and O- images with a nucleus in the center were observed from the Alq3 side. It is concluded from the results that the constituent element Mg of the cathode was oxidized at the interface adjacent to the Alq3 layer during exposure to the atmosphere, forming a dark spot with a nucleus in the center. Finally, it was confirmed that the TOF-SIMS analysis of the tape-stripped surface is useful for the analysis of the mechanism of dark spot formation. PMID- 11393849 TI - Utilization of kinetically enhanced monovalent binding affinity by immunoassays based on multivalent nanoparticle-antibody bioconjugates. AB - The monovalent binding affinity of high binding site density nanoparticle antibody bioconjugates is shown to exceed the intrinsic affinity of the original, monoclonal antibody. The nanoparticle-antibody bioconjugates were prepared by covalent coupling of antibodies to long-lifetime fluorescent, europium(III) chelate nanoparticles, 107 nm in diameter. Experiments were carried out in standard microtitration wells to determine solid-phase association and dissociation rate constants, nonspecific binding, and affinity constants of the various binding site density nanoparticle-antibody bioconjugates and the conventionally labeled monoclonal antibody. The affinity constant for monovalent binding of a high binding site density bioconjugate (5.4 x 10(10) M(-1)) was 8 fold higher than the intrinsic affinity of the antibody (6.6 x 10(9) M(-1)). The separately measured association (2.5 x 10(6) M(-1) s(-1)) and dissociation (3.7 x 10(-5) s(-1)) rate constants of the bioconjugate were 2-fold higher and 4-fold lower, respectively, compared to the antibody. The dependence of the association rate constant of the density of the binding sites enhanced the kinetics and the affinity of the high binding site density bioconjugates. The nanoparticle labels with high specific activity, low nonspecific binding, and enhanced binding affinity of the nanoparticle-antibody bioconjugates contribute to the design of the next generation immunoassays with extreme sensitivity. PMID- 11393850 TI - Scanning electrochemical microscopy. 41. Theory and characterization of ring electrodes. AB - Ring ultramicroelectrodes, which are of particular interest as probes for scanning electrochemical microscopy (SECM), combined with near-field scanning optical microscopy, were investigated. Theoretical SECM tip current-distance (approach) curves for a ring electrode were calculated by numerical (finite element) analysis. The SECM curves obtained were a function of the geometry of the tips including the thickness of the ring and the insulating sheath. Theoretical approach curves over conductive substrates showed a strong dependence on the ratio of inner to outer radii of ring microelectrodes (a/b) and were relatively insensitive to the thickness of the insulating sheath (r(g)). For insulating substrates, however, the approach curves varied significantly with r(g), but much less with the a/b ratio. Comparison of experimental and theoretical SECM curves provided a good method of evaluating the size and shape of ring electrodes. Good agreement of the experimental and theoretical curves was found with a ring microelectrode with a nominal 200-nm ring thickness, yielding values of 1.7, 1.9, and 5.7 microm for the inner (a) and outer (b) radii of a ring and the outermost radius of insulating sheath (r(g)), respectively. PMID- 11393851 TI - Simultaneous multianalyte detection with a nanometer-scale pore. AB - It was recently shown that naturally occurring, genetically engineered or chemically modified channels can be used to detect analytes in solution. We demonstrate here that the overall range of analytes that can be detected by single nanometer-scale pores is expanded using a potentially simpler system. Instead of attaching recognition elements to a channel, they are covalently linked to polymers that otherwise thread through a nanometer-scale pore. Because the rate of unbound polymer entering the pore is proportional to its concentration in the bulk, the binding of analyte to the polymer alters the latter's ability to thread through the pore, and the signal that results from individual polymer translocation is unique to the polymer type; the method permits multianalyte detection and quantitation. We demonstrate here that two different proteins can be simultaneously detected with this technique. PMID- 11393852 TI - Individually addressable gel-integrated voltammetric microelectrode array for high-resolution measurement of concentration profiles at interfaces. AB - The application of a novel voltammetric probe, based on an individually addressable gel-integrated microelectrode array (IA-GIME), for real-time, high spatial resolution concentration profile measurements at interfaces is described. Reliability and validity of steep metal concentration gradients obtained with this novel system have been demonstrated by performing systematic tests at well controlled liquid-liquid and liquid-solid interfaces. The liquid-liquid interface was formed by two layers of aqueous solutions with different components; only one layer contained trace metal ions (Pb(II) and Cd(II)); the individually addressable microelectrode array was placed at the interface of the liquid-liquid system; the concentration profiles were recorded as function of time; and the effective diffusion coefficients were calculated. The liquid-"solid" interface was formed from an aqueous solution layer overlying a bed of silica particles saturated with an aqueous solution. The sensor array has been used to monitor the diffusion processes of Tl(I) or Pb(II) from the liquid phase to the "solid" phase. The influences of porosity, geometry of the porous media, and complexation between metal ion and silica, on the diffusion processes, have been studied. All these results show that correct diffusion profiles of metal ions at interfaces can be obtained with 200-microm resolution with the IA-GIME. They also demonstrate that, for measurements in "solid" phase, the aforementioned factors must be considered carefully for correct calibration of any electrodes and the gel-integrated microelectrodes are unique tools to enable calibration of the sensors with synthetic solutions. PMID- 11393853 TI - Hydrodynamic potential-modulated reflectance spectroscopy: theory and experiment. AB - This article describes the development and application of a new electrochemical methodology based on potential-modulated UV-vis reflectance spectroscopy (PMRS). The device configuration is based upon a thin-layer flow-through channel cell incorporating a platinum working electrode. Reagent solutions are pumped through the cell under well-defined hydrodynamic conditions and electrolyzed at the platinum working electrode. Measurements are presented for linear sweep and fixed dc potentials with a superimposed small amplitude sinusoidal potential perturbation. A UV-vis source is employed to irradiate the electrode region, and the resulting reflected signal is analyzed using a phase sensitive detector. Experimental studies using tris(4-bromophenyl) amine (TBPA) in acetonitrile are presented which quantify the relationship between the absorption spectrum and reflected light intensity as a function of the transport rate, electrolysis reactions, and the modulation frequency of the incident irradiation. The experimental results are analyzed using numerical simulations based on a finite difference strategy. These permit the quantitative prediction of the concentration distribution of reagents within the cell. A fast Fourier transform (FFT) routine was used to analyze the frequency response of the numerically predicted reflectance signal. Excellent agreement was observed between the numerical predictions and experimental observations. PMID- 11393854 TI - Existence of phase explosion during laser ablation and its effects on inductively coupled plasma-mass spectroscopy. AB - A sudden increase in crater depth was observed during high irradiance (> 10(10) W/cm2) laser ablation of silicon, and it is attributed to the phenomenon of phase explosion. The threshold irradiance for phase explosion showed a dependence on two laser parameters: laser beam spot size and wavelength. For a larger beam size and longer incident wavelength, a higher laser irradiance was required to generate phase explosion. The rapid increase of crater depth above the phase explosion threshold irradiance correlated with a significant increase in the ICPMS signal intensity. The ratio of crater volume to ICPMS intensity, which represents entrainment efficiency, remained the lowest at laser irradiances slightly above the phase explosion threshold. However, this ratio increased at irradiances well above the threshold (> 10(11) W/cm2). Chemical analysis using laser ablation at irradiance above 10(11) W/cm2 provides increased sensitivity via improved entrainment and transport efficiency and increased ablation rate. PMID- 11393855 TI - Statistical two-dimensional correlation spectroscopy: its theory and applications to sets of vibrational spectra. AB - The present study aims at developing a new form of two-dimensional (2D) correlation spectroscopy, statistical 2D correlation spectroscopy. Statistical 2D spectroscopy differs from the widely used generalized 2D correlation spectroscopy in that the former abstracts spectral features by pretreatment and by 2D maps that are limited by the correlation coefficients in the range from 1 to -1. In this paper, the theory of the new 2D method is briefly described, and then its applications are discussed to reveal spectral and concentration features of artificial model spectra, infrared spectra of polycondensation of bis(hydroxyethyl terephthalate) measured on-line, and short-wave near-infrared spectra of raw milk. The results are analyzed thoroughly and compared with those from generalized 2D correlation spectroscopy and partial least-squares loadings and scores. The most significant advantage of statistical 2D correlation spectroscopy is that the 2D correlation spectra are easy to calculate and are purely mathematical in nature, thereby eliminating any subjective involvement of an experimenter, while the inherent weakness of the method lies in its sensitivity to the noise. PMID- 11393856 TI - Two-dimensional fluorescence correlation spectroscopy with modulated excitation. AB - Overlap of multiple states or multiple species in a chemical system often creates a congested fluorescence spectrum that is difficult to interpret. The resolution of component spectra is essential for the understanding of the structure and dynamics of such multicomponent systems. In this paper, two-dimensional fluorescence correlation spectroscopy (2D FCS) is presented for the dissection of component spectra using the time correlation function. In 2D FCS, the time response of fluorescence intensity is collected at various wavelengths upon an external perturbation. The time correlation function is evaluated between wavelengths. A two-dimensional fluorescence correlation spectrum, or a plot of the correlation intensity as a function of two wavelength axes, resolves the overall spectrum into component spectra. The characteristics of the two dimensional time correlation function are demonstrated in the frequency domain fluorescence spectroscopy in which the sinusoidally modulated excitation provides the external perturbation. Using 2D FCS, fine vibronic structures of the component fluorescence emission spectra were completely resolved from a strongly overlapped one-dimensional mixture spectrum. The existence of multiple microenvironments of a probe molecule in a biological system is evidenced by nonzero asynchronous correlation intensities. The corresponding spectra are retrieved from correlation analysis. Unlike traditional resolution methods in fluorescence spectroscopy based on statistical fitting of fluorescence decays, 2D FCS can resolve species whose fluorescence decays are linked by the rate constants in chemical reactions and species displaying multiexponential decay kinetics. PMID- 11393857 TI - Determination of the primary structures of heparin- and heparan sulfate-derived oligosaccharides using band-selective homonuclear-decoupled two-dimensional 1H NMR experiments. AB - Band-selective homonuclear-decoupled (BASHD) two-dimensional NMR experiments are applied to the assignment of 1H NMR spectra of oligosaccharides, using as an example a heparin-derived hexasaccharide. The anomeric (H1) region of the 1H NMR spectrum is band-selected in the F1 dimension. With the increased resolution that results from less truncation of interferograms in the t1 dimension, finer digital resolution in the F1 dimension, and collapse of multiplets to singlets in the F1 dimension, cross-peaks to the anomeric protons of the two iduronic acid residues, which overlap in normal two-dimensional total correlation spectroscopy (TOCSY) and rotating frame Overhauser enhancement spectroscopy (ROESY) spectra of the hexasaccharide, are resolved in BASHD-TOCSY and BASHD-ROESY spectra, leading to an unequivocal assignment of the 1H NMR spectrum of the hexasaccharide. Incorporation of the water attenuation by transverse relaxation method for the complete and selective elimination of the water resonance into two-dimensional BASHD experiments makes it possible to observe oligosaccharide resonances at the frequency of the water resonance, as demonstrated with the observation of cross peaks to resonances at the frequency of the water resonance in BASHD-TOCSY spectra of a second heparin-derived hexasaccharide. PMID- 11393858 TI - Quantitative detection of aromatic compounds in single aerosol particle mass spectrometry. AB - Most laser-based aerosol mass spectrometers rely on a single ultraviolet laser to both ablate and ionize the aerosol particle. This technique produces complex and fragmented mass spectra, especially for organic compounds. The approach presented here achieves a more robust and quantitative analysis using a CO2 laser to evaporate the aerosol particle and a vacuum ultraviolet laser to ionize the vapor plume. Vacuum ultraviolet laser ionization produces little fragmentation in the mass spectra, making the identification of an aerosol particle's constituents more straightforward. An analysis of simple, three-component mixtures of aniline, benzyl alcohol, and m-nitrotoluene shows that the technique also provides a quantitative analysis for all the components of the mixture. Furthermore, the detection of predominantly parent ion signal from anthracene particles demonstrates the utility of the technique in the analysis of lower vapor pressure, solid-phase aerosols. Finally, we discuss the potential and limitations of this technique in analyzing organic atmospheric aerosols. PMID- 11393859 TI - Structural characterization of Mycobacterium tuberculosis lipoarabinomannans by the combination of capillary electrophoresis and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) associated with capillary electrophoresis (CE) has been used for structural characterization of mannooligosaccharide caps from Mycobacterium tuberculosis H37rv mannosylated lipoarabinomannans (ManLAMs). The mannooligosaccharide caps were released by mild acid hydrolysis, labeled with 1 aminopyrene-3,6,8-trisulfonate (APTS) prior to being separated by CE, collected, and analyzed by MALDI-TOF-MS and post-source decay experiments. This approach was optimized using standard APTS-labeled oligosaccharides. With the selected (9:1) mixture of 2,5-dihydroxybenzoic acid (DHB) and 5-methoxysalicylic acid (MSA) as matrix and the on-probe sample cleanup procedure with cation-exchange resin, standard APTS-maltotriose was successfully detected down to 50 fmol using linear mode negative MALDI-TOF-MS. Moreover, using extraction delay time, only 100 and 500 fmol of this standard were required, respectively, to obtain accurate reflectron mass measurements and sequence determination through post-source decay experiments. Applied to only 5 microg (294 pmol) of M. tuberculosis ManLAMs, this analytical approach allowed successful mass characterization of the mannooligosaccharide cap structures from the deprotonated molecular ions [M - H]- and the y-type ion fragments obtained in post-source decay experiments. This powerful analytical approach opens new insights into both the characterization of oligosaccharides and the capping motifs displayed by ManLAMs purified from mycobacteria isolated from tubercular patients without in vitro culturing. PMID- 11393860 TI - Discrimination between bacterial spore types using time-of-flight mass spectrometry and matrix-free infrared laser desorption and ionization. AB - We demonstrate that molecular ions with mass-to-charge ratios (m/z) ranging from a few hundred to 19 050 can be desorbed from whole bacterial spores using infrared laser desorption and no chemical matrix. We have measured the mass of these ions using time-of-flight mass spectrometry and we observe that different ions are desorbed from spores of Bacillus cereus, Bacillus thuringiensis, Bacillus subtilis, and Bacillus niger. Our results raise the possibility of identifying microorganisms using mass spectrometry without conventional sample preparation techniques such as the addition of a matrix. We have measured the dependence of the ion yield from B. subtilis on desorption wavelength over the range 3.05-3.8 microm, and we observe the best results at 3.05 microm. We have also generated mass spectra from whole spores using 337-nm ultraviolet laser desorption, and we find that these spectra are inferior to spectra generated with infrared desorption. Since aerosol analysis is a natural application for matrix free desorption, we have measured mass spectra from materials such as ragweed pollen and road dust that are likely to form a background to microbial aerosols. We find that these materials are readily differentiated from bacterial spores. PMID- 11393861 TI - Application of the ART-2a algorithm to laser ablation aerosol mass spectrometry of particle standards. AB - Single-particle mass spectrometers are now commonly used to analyze atmospheric particles and generate tens of thousands of spectra from typical measurement campaigns. The ART-2a spectrum algorithm has been used to classify these spectra. In this work, we generate a range of particles that are models of those that are common in the atmosphere. A single-particle mass spectrometer is used to analyze these known particles, and the spectra are classified using ART-2a. The optimum vigilance parameter is approximately 0.5 while the optimum learning rate is approximately 0.05. The classifications elucidate limitations in generation of test particles, their analysis by single-particle techniques, and their classification by ART-2a. PMID- 11393863 TI - Electroosmotic flows in microchannels with finite inertial and pressure forces. AB - Emerging microfluidic systems have spurred an interest in the study of electrokinetic flow phenomena in complex geometries and a variety of flow conditions. This paper presents an analysis of the effects of fluid inertia and pressure on the velocity and vorticity field of electroosmotic flows. In typical on-chip electrokinetics applications, the flow field can be separated into an inner flow region dominated by viscous and electrostatic forces and an outer flow region dominated by inertial and pressure forces. These two regions are separated by a slip velocity condition determined by the Helmholtz-Smoulochowski equation. The validity of this assumption is investigated by analyzing the velocity field in a pressure-driven, two-dimensional flow channel with an impulsively started electric field. The regime for which the inner/outer flow model is valid is described in terms of nondimensional parameters derived from this example problem. Next, the inertial forces, surface conditions, and pressure-gradient conditions for a full-field similarity between the electric and velocity fields in electroosmotic flows are discussed. A sufficient set of conditions for this similarity to hold in arbitrarily shaped, insulating wall microchannels is the following: uniform surface charge, low Reynolds number, low Reynolds and Strouhal number product, uniform fluid properties, and zero pressure differences between inlets and outlets. Last, simple relations describing the generation of vorticity in electroosmotic flow are derived using a wall-local, streamline coordinate system. PMID- 11393862 TI - Geometric scaling effects in electrical field flow fractionation. 1. Theoretical analysis. AB - This work outlines the fundamental scaling laws associated with electrical field flow fractionation channels. Although general FFF theory indicates few advantages from miniaturization, EFFF theory indicates clear advantages to miniaturization of the EFFF channel. Retention, plate heights, resolution, equilibration times, and time constants are examined. The outlined theory predicts scaling advantages in each of these areas after miniaturization. Potential applications, such as the use of these systems for sample preparation in microscale total analysis systems, and improvements associated with these theoretical predictions are also discussed. PMID- 11393864 TI - A high-performance and simplified quasi-elastic laser scattering method using homodyne detection in beam divergence. AB - We devise the new principle of the quasi-elastic laser scattering (QELS) method using a homodyne detection technique in a beam divergence and successfully facilitate the equipment The QELS method is a unique technique for the noncontact and time-resolved study of surface tension at liquid surfaces and liquid/liquid interfaces. The conventional QELS method requires a precise optical alignment using a local oscillator such as a diffraction grating, and the determination of the surface tension accompanies much difficulty because of the low S/N ratio of the power spectra. Our new principle allows high-performance QELS measurements by only a simple alignment of a downsized experimental setup. The power spectra are obtained with 50-100 times higher S/N ratios than the conventional ones. The power spectra are analyzed by a new theory, and the calculated surface tensions agree with the literature values. The accuracy of the surface tension measurements using the QELS method is substantially improved. PMID- 11393865 TI - Ets Gene Family. PMID- 11393866 TI - Are basic assumptions correct--is endometriosis a progressive, self-destructive disease? PMID- 11393867 TI - Design and Analysis of Cluster Randomized Trials. Workshop proceedings. University of Sheffield, 5-7 July 1999. PMID- 11393868 TI - Control of a mucosal challenge and prevention of AIDS by a multiprotein DNA/MVA vaccine. AB - Heterologous prime/boost regimens have the potential for raising high levels of immune responses. Here we report that DNA priming followed by a recombinant modified vaccinia Ankara (rMVA) booster controlled a highly pathogenic immunodeficiency virus challenge in a rhesus macaque model. Both the DNA and rMVA components of the vaccine expressed multiple immunodeficiency virus proteins. Two DNA inoculations at 0 and 8 weeks and a single rMVA booster at 24 weeks effectively controlled an intrarectal challenge administered 7 months after the booster. These findings provide hope that a relatively simple multiprotein DNA/MVA vaccine can help to control the acquired immune deficiency syndrome epidemic. PMID- 11393869 TI - Salt and blood pressure. Conventional wisdom reconsidered. AB - The salt hypothesis is that higher levels of salt in the diet lead to higher levels of blood pressure, increasing the risk of cardiovascular disease. Intersalt, a cross-sectional study of salt levels and blood pressures in 52 populations, is often cited to support the salt hypothesis, but the data are somewhat contradictory. Four of the populations (Kenya, Papua, and 2 Indian tribes in Brazil) do have low levels of salt and blood pressure. Across the other 48 populations, however, blood pressures go down as salt levels go up, contradicting the hypothesis. Experimental evidence suggests that the effect of a large reduction in salt intake on blood pressure is modest, and health consequences remain to be determined. Funding agencies and medical journals have taken a stronger position favoring the salt hypothesis than is warranted, raising questions about the interaction between the policy process and science. PMID- 11393870 TI - Gatekeepers and sentinels. Their consolidated effects on inpatient medical care. AB - Evaluations assessing precertification by nurse gatekeepers and onsite monitoring by nurse sentinels report inconclusive unique effects of these programs on the utilization, expense, and appropriateness of inpatient medical care. By applying the fixed- and random-effects paradigm of meta-analysis, this article consolidates the results of all relevant quasi-experiments conducted by an evaluation group of a large private insurer from 1986 to 1990. It determines the difference in effect between the target and comparison groups, reports this effect and its statistical range, and determines the pooled effect and its range. The random effects indicate that precertification will reduce admissions, and onsite, concurrent review will reduce length of stay, bed days, and inpatient ancillary expense. The precertification and onsite programs may reduce negative iatrogenic effects, thereby enhancing the patients' well-being. If applied to privately insured populations who are still served on a fee-for-service basis, the gatekeeper and sentinel effects of these programs may reduce utilization and expense; however, inference of these results to Medicare fee-for-service care remains problematical. PMID- 11393872 TI - Changing work ethic and welfare dependence through welfare reform. The 100-hour rule waiver experiment for AFDC-U. AB - Assumptions about welfare dependency and work are examined in a randomized experiment that tested the impact of the 100-hour rule waiver for two-parent welfare families: Aid to Families with Dependent Children-Unemployed (AFDC-U). The 100-hour rule was waived for the experiment group, allowing the primary wage earners in these families to work more than 100 hours a month without losing welfare eligibility. The analysis uses county administrative data, unemployment insurance records, and Medicaid data, and compares regression adjusted least squares means for the control and experiment group. The results from the impact analysis indicate that waiving the 100-hour rule has no effect on primary wage earners' work activity and earnings. The 100-hour rule waiver also has little effect on time on aid and AFDC-U payments, and does not have an effect in reducing marital dissolution. The results cast doubts about the validity of the assumptions underlying some of the recent welfare reform initiatives. PMID- 11393871 TI - Alternative solutions to the problem of selection bias in an analysis of federal residential drug treatment programs. AB - In an evaluation of prison-based residential drug treatment programs, the authors use three different regression-based approaches to estimating treatment effects. Two of the approaches, the instrumental variable and the Heckman approach, attempt to minimize selection bias as an explanation for treatment outcomes. Estimates from these approaches are compared with estimates from a regression in which treatment is represented by a dummy variable. The article discusses the advantage of using more than one method to increase confidence in findings when possible selection bias is a concern. Three-year outcome data for 2,315 federal inmates are used in analyses where the authors separately examine criminal recidivism and relapse to drug use for men and women. Statistical tests lead the authors to conclude that treatment reduces criminal recidivism and relapse to drug use. The treatment effect was largest when the inference was based on the Heckman approach, somewhat smaller when based on the instrumental variable approach, and smallest when based on the traditional dummy variable approach. Treatment effects for females were not statistically significant. PMID- 11393873 TI - Rasch calibration and optimal categorization of an instrument measuring women's exercise perseverance and barriers. AB - The purposes of this study were: (a) to determine the optimal categorization of an instrument measuring women's exercise perseverance and barriers using the Rasch analysis and (b) to examine urban women's exercise perseverance and barriers. A 23-item barrier instrument with five response categories was administered to 479 women from a metropolitan area. The data analysis started from collapsing the original five adjacent categories into three and four categories and 11 sets of original and collapsed data were analyzed using the Rasch rating scale model. The model-data fit, category and separation statistics, and parameter estimates provided by Rasch analyses were used to determine the optimal categorization of the instrument. Instead of the original five-category construct, which had a disordered internal construct, a collapsed three-category construct (i.e., Very Often/Often, Sometimes/Rarely, and Never) was found to have the optimal categorization. The time barrier domain was found to be the most severe barrier domain, but the barrier "lack of self-discipline" was the most severe individual barrier. Rasch calibration provides a new way to construct an instrument with optimal categorization, to describe the nature of barrier items and the respondents' attribute being measured, and to develop a practical and informative scoring sheet. PMID- 11393874 TI - Psychometric properties of child- and teacher-reported curl-up scores in children ages 10-12 years. AB - The purpose of this study was to examine the psychometric properties of child- and teacher-reported curl-up (CU) scores in children ages 10-12 years in both a norm-referenced (NR) and criterion-referenced (CR) framework. Eighty-four children, 36 boys and 48 girls, performed the FITNESSGRAM (Cooper Institute for Aerobics Research, 1992) CU test on 2 days separated by 48-72 hr. Two video cameras were used to record students' CU performances. Two students performed the CU at the same time, with each child's performance recorded by one camera. The test was terminated when the child stopped due to fatigue or after two form errors occurred. Teacher-reported scores were the average of two independent ratings of each video performance, while child-reported scores came from data collected and recorded by the children. Single trial norm-referenced reliability was R = .75 for girls and R = .80 for boys for teacher-reported CU and R = .69 and R = .70 for child-reported CU for girls and boys, respectively. CR reliability was examined using P, proportion of students who consistently passed or failed the test across 2 days, and km, defined as reliability with chance removed. For teacher-reported scores, P = .89 and km = .78 for boys and P = .81 and km = .62 for girls. For child-reported scores, P = .86 and km = .72 for boys, while P = .79 and km = .58 for girls. For teacher-reported data, 39% of boys passed and 50% failed the test on both days, while for girls the percentages were 27% pass and 54% fail. For child-reported data, 64% of boys passed and 22% failed on both days, while 54% of girls passed and 25% failed. NR validity was examined by correlating teacher and child-reported scores. The resultant coefficient was r = .42 (95% CI = .11-.66) for boys and r = .67 (95% CI = .58-.74) for girls. Additionally, child-reported scores were significantly higher than teacher reported scores. CR validity was examined with a contingency coefficient, and results indicated C = .55 with 44% false master errors for boys and C = .65 with 29% false master errors for girls. The findings of this study suggest that while NR reliability estimates were moderate for teacher-reported scores, single trial estimates suggest that child-reported CU should be viewed with caution. In regard to CR reliability, both teacher-reported and child-reported reliability were moderate. However, there were marked differences between teacher- and child reported scores, with children reporting higher percentages of students passing and lower percentage of student failing the test when compared with scores reported by teachers. Validity was rather moderate when viewed in either a NR and CR framework. It is suggested that problems with child-reported scores may be due to the need for additional practice or simplification of the testing protocol. PMID- 11393875 TI - The most important points in grand slam singles tennis. AB - A computerized data management system was used to enter details of points played in 252 tennis matches from the men's and women's singles events of all four Grand Slam tournaments over a 2-year period. A supplementary data analysis system was developed to determine the proportion of points won by each player on serve at each game score from love all to deuce as well as the proportion of games the player went on to win from each score. Analysis of the 43 matches in which both players served at each score from love all to deuce revealed that the proportion of points won by the server was not significantly influenced by score, F(15, 495) = 0.8, p > .05. A further analysis of the 175 matches consisting of at least 100 points revealed that the proportion of points won by the superior player was not significantly influenced by gender, F(1, 165) = 0.1, p > .05, or surface, F(3, 165) = 0.1, p > .05. However, the proportion of points won when serving was significantly greater in men's singles than women's singles, F(1, 165) = 69.7, p < .001, R2 = .30. Surface also had a significant influence on the proportion of points won when serving, F(3, 165) = 8.1, p < .001, R2 = .13, with a significantly greater proportion of points won when serving by both winning and losing players at Wimbledon than at the Australian and French Opens, p < .05. This suggests that gender and surface should be accounted for when determining the importance of points in Grand Slam tennis. PMID- 11393876 TI - Learning a coordination skill: interactive effects of instruction and feedback. AB - Information prior to and during the acquisition of a continuous bimanual task was manipulated. Participants practiced a difficult coordination pattern, which produced circular shapes on the computer, when they moved their arms correctly. Four groups were examined, which differed in the type and amount of information provided. Either limb or circle feedback was provided in the presence or absence of instructions detailing how to move the limbs. Circle feedback facilitated learning relative to the limb feedback in which the explicit displacements of the limbs was displayed. Under circle feedback conditions, instructions hindered acquisition. Little instructional effects were observed under limb feedback conditions, despite the prediction that instructions would benefit learning when the feedback was more compatible. Findings are discussed in relation to the complexity of the feedback and processing demands. PMID- 11393877 TI - On the tactical significance of game situations in anticipating ball trajectories in tennis. AB - It is hypothesized that live playing situations preceding an opponent's strokes in tennis have sufficient significance to provide expert players with anticipative cues to estimate accurately the spatiotemporal characteristics of oncoming ball trajectories. Seven participants (all expert tennis players) had to watch two players opposed in high, moderate, and low tactical significance situations terminated by a stroke delivered by one of the two players in the direction of the participants. The participants' vision was occluded 100 ms after the stroke, and they had to indicate the zone reached by the ball at the moment of its rebound. Results showed that the essential anticipative information is contained in the view of the opponent's stroke movements, whatever the tactical significance of the situation. PMID- 11393878 TI - Situational interest in physical education: a function of learning task design. AB - Situational interest is the appealing effect of unique characteristics students recognize in a learning task during interaction with the task. It occurs when a learning task gives the learner a sense of novelty and challenge, demands high attention and exploration intention, and generates instant enjoyment during the person-task interaction. In this study, a repeated measure research design was used to examine the effects of task design on situational interest and the extent to which the effects were mediated by gender, grade, personal interest, and skill levels. Middle school students (N = 242) evaluated situational interest of four learning tasks with different cognitive and physical demands after having experienced the tasks in their physical education classes. Analyzed data showed that cognitive demand of a learning task played a critical role in generating situational interest. Grade levels, gender, and personal interest mediated the effects of task design on situational interest. But these mediation effects seemed rather limited. Physical skill levels had little influence on the effects of task design on situational interest. The findings seem to suggest that to enhance interestingness of a physical activity task, an option for physical educators may be to increase cognitive demand rather than reduce physical demand. PMID- 11393879 TI - Students' perceptions of the motivational climate, achievement beliefs, and satisfaction in physical education. AB - The present study examined the relationship between students' perceptions of the motivational climate and beliefs about the causes of success, preference for challenging tasks, and satisfaction in physical education. Responses of 50 female and 46 male students (M age = 12.08 years; SD = .72) showed that perceptions of a mastery-oriented motivational climate were related to the belief that motivation or effort caused success and satisfaction. In contrast, perceptions of a performance climate were related to the belief that deception caused success and related negatively to the students' preference for challenging tasks. Results of hierarchical regression analyses revealed that perceptions of the motivational climate explained a significant amount of unique variance in the students' responses after controlling for dispositional goal orientations. The results suggest that the teacher can influence the salience of a mastery-oriented climate and, in so doing, optimize a child's motivation in physical education. PMID- 11393880 TI - Energy expenditure relative to perceived exertion: stationary cycling versus treadmill walking. PMID- 11393881 TI - Relationship of blood cholesterol to body composition, physical fitness, and dietary intake measures in third-grade children and their parents. PMID- 11393882 TI - The effects of the environment on physical activity patterns of children with mental retardation. PMID- 11393883 TI - Perceived physical ability as a function of race and racial comparison. PMID- 11393884 TI - Predicting children's overarm throw ball velocities from their developmental levels in throwing. AB - This study examined the movement process-product relationship from a developmental perspective. The authors used multiple regression to investigate the changing relationship between qualitative movement descriptions of the overarm throw and the throwing outcome, horizontal ball velocity. Seventeen girls and 22 boys were filmed longitudinally at ages 6, 7, 8, and 13 years. Their movements were assessed using Roberton's (Roberton & Halverson, 1984) developmental sequences for action of the humerus, forearm, trunk, stepping, and stride length. The sequences accounted for 69-85% (adjusted) of the total velocity variance each year. The components that best predicted ball velocity changed over time, although humerus or forearm action always accounted for considerable variance. Gender was a good predictor of ball velocity, but if the developmental descriptions were entered first in a stepwise regression, gender then explained no more than 2% additional variance. PMID- 11393885 TI - Involvement of p53-dependent apoptosis in radiation teratogenesis and in the radioadaptive response in the late organogenesis of mice. AB - The irradiation of fetuses at the late period of organogenesis has been known to induce a dramatic increase in malformations. The mechanisms involved, however, have remained unclear for a long time. Using the mouse limb bud system, we first found that radiation-induced apoptosis is involved in the malformation, namely, radiation-induced apoptosis in the predigital regions of embryonic limb buds is responsible for digital defects in mice. An examination of embryonic C57BL/6J mice with different p53 (trp53) status enabled us to further find that susceptibility to radiation-induced apoptosis in the predigital regions and digital defects depend on both the p53 status and the radiation dose; p53 wild type mice appeared to be the most sensitive, while p53 knockout mice were the most resistant. These results indicate that p53-dependent apoptosis mediates radiation-induced digital defects in the later organogenesis period. The existence of a radioadaptive response in embryonic mice, which has not been reported so far, was found by irradiating embryos with either 5 cGy or 30 cGy on embryonic day 11 prior to a challenging irradiation at 3 Gy on embryonic day 12. p53-heterozygous embryos did not show the radioadaptive response, indicating the involvement of p53 in the radioadaptive response in embryogenesis. PMID- 11393886 TI - Role of the Escherichia coli and human DNA glycosylases that remove 5 formyluracil from DNA in the prevention of mutations. AB - Ionizing radiation induces a wide variety of modifications to purine and pyrimidine residues. The exocyclic methyl group of thymine does not escape oxidative damage. Any 5-hydroperoxymethyluracil produced is spontaneously decomposed to form 5-formyluracil (5-foU) and 5-hydroxymethyluracil. The yield of 5-foU by ionizing radiation is roughly the same as that of 8-oxoguanine. 5-foU is a potential mutagenic damage in vitro and in vivo. Mammalian cells have an activity that removes 5-foU from X-irradiated DNA. Furthermore, the Nth, Nei and MutM proteins of E. coli have DNA glycosylase/AP lyase activities that recognize and remove 5-foU in DNA. The mutation frequency of 5-foU-containing plasmid increases when replicated in E. coli nthneimutMalkA. Single mutations in the nth, nei or mutM gene do not affect the mutation frequency. Therefore, these gene products are likely backup enzymes used to repair 5-foU in DNA. Furthermore, the human hNTH1 enzyme, a homologue of E. coli Nth, is found to have similar DNA glycosylase activity to that of the Nth protein. PMID- 11393887 TI - Radioprotectors in radiotherapy. PMID- 11393888 TI - Treatment planning for heavy-ion radiotherapy: biological optimization of multiple beam ports. AB - A crucial task in radiotherapy is dose conformation to the prescribed target volume whilst sparing the surrounding healthy tissue around as much as possible. One of the best approaches so far is active dose shaping in three dimensions using scanned beams of charged particles, like carbon ions. Besides their inverse dose profile and minimal lateral scattering, carbon ions have the advantage that their RBEs increase towards the end of their range. An active beam-delivery system for intensity-modulated carbon-ion beams has been operational at GSI since December, 1997. In order to ensure dose conformation, inverse treatment planning with respect to the biologically effective dose distribution must be applied. A typical patient irradiation comprises two singly optimized opposing fields. This paper discusses the superposition of biologically effective dose distributions for radiotherapy with 12C ions, which is non-trivial due to the nonlinear nature of the dose response of biological systems. Sum rules for the nonlinear addition of singly optimized fields are derived. This method is being used clinically, and has been successfully applied to more than 50 patients. PMID- 11393889 TI - Directional distribution of radiation around an accident at a uranium fuel factory in Tokai-mura, 1999. AB - A beta-ray survey was carried out on concrete walls of the boundary and buildings after a criticality accident at a factory of JCO Co. Ltd. at Tokai-mura. A remarkable distribution of beta counts was observed on the walls depending on the complex internal and external structures of buildings surrounding a precipitation vessel containing uranium 23 days after the accident. The directional distribution function, based on the beta counts on the walls, was consistent with data concerning the neutron dose rate measured in several directions during the accident, suggesting an anisotropic neutron distribution to the residential area. PMID- 11393890 TI - Cosmic radiation protection dosimetry using an Electronic personal Dosemeter (Siemens EPD) on selected international flights. AB - The effectiveness of an Electronic Personal Dosemeter (Siemens EPD) for cosmic radiation dosimetry at aviation altitudes was examined on eight international flights between March and September, 1998. The EPD values (Hepd) of the dose equivalent from penetrating radiation, Hp(10), were assumed to be almost the same as the electron absorbed doses during those flights. Based on the compositions of cosmic radiation in the atmosphere and the 1977 ICRP recommendation, an empirical equation to conservatively estimate the personal dose equivalent (Hp77) at a depth of 5 cm was derived as Hp77 = 3.1 x Hepd. The personal dose equivalent (Hp90) based on the 1990 ICRP recommendation was given by Hp90 = 4.6 x Hepd; the conservative feature of Hp90 was confirmed in a comparison with the calculated effective doses by means of the CARI-6 code. It is thus expected that the EPD will be effectively used for radiation protection dosimetry on selected international flights. PMID- 11393891 TI - Glow-peak stability in 6LiF:Mg,Ti (TLD-600) exposed to a Fe-ion beam. AB - The stability of glow peaks in 6LiF:Mg,Ti (TLD-600) exposed to a high-energy Fe ion beam was examined in comparison to 137Cs gamma-ray irradiation under changing annealing conditions. The peak areas induced by the Fe ions were much smaller than those by gamma-rays. The sizes and positions of peaks 3-5 in Fe-ion irradiated samples were hardly changed after post-annealing at 100 degrees C x 30 min, regardless of the pre-annealing conditions (fast quenching or subsequent pre annealing at 100 degrees C x 2 h). Whereas, the peaks in gamma-ray irradiated samples were notably affected by post-annealing; the peak positions and peak-area sizes changed in different ways depending on the pre-annealing conditions. The effects of post-annealing on peak 6 were identical for Fe ions and gamma-rays. These facts suggest that peaks 3-5 in TLD-600 comprised both stable and unstable luminescent centers, and that the latter part would be easily depleted in highly dense ionization. PMID- 11393892 TI - Relative biological effectiveness of the 235 MeV proton beams at the National Cancer Center Hospital East. AB - A therapy-dedicated cyclotron was installed in the National Cancer Center Hospital East (NCCHE) at Kashiwa in 1997. Prior to the start of clinical use, we investigated the biological effectiveness of therapeutic proton beams for cell lethality. The proton beams accelerated up to 235 MeV were horizontally extracted from the cyclotron, and scattered by a bar-ridge filter to produce a Spread-Out Bragg-Peak (SOBP) of 10-cm width. The biological systems used here were mouse intestinal crypt cells and three in vitro cell lines, including SCC61 human squamous cell carcinoma, NB1RGB human fibroblasts and V79 Chinese hamster cells. The dose responses after irradiation at either the entrance plateau or the middle portion of SOBP were compared with those after linac 6 MV X-ray irradiation. The fit of a linear quadratic model to survival curves showed that proton irradiation increased the alpha value of SCC61 and the beta value of V79 cells with a least change for alpha/beta ratio of NB1RGB cells. The isoeffect dose that reduces either cell survivals to 10% or mouse jejunum crypts to 10 per circumference was termed D10. The relative biological effectiveness (RBE) of protons obtained by comparing the D10 values between protons and X-rays ranged from 0.9 to 1.2. The depth distribution of cell lethality was measured by replating V79 cells after irradiation from a "cell stack chamber" that received a single dose of 7 Gy at the middle position of SOBP. The thus-obtained cell survivals at various depths coincided well with the estimated survivals, but tended to decrease at the distal end of SOBP. We conclude that an RBE of 1.1 would be appropriate for 235 MeV proton beams at the NCCHE. PMID- 11393893 TI - Neutron-induced adaptive response studied in go human lymphocytes using the comet assay. AB - This study demonstrates that cells adapted to ionizing radiation developed reduced initial DNA damage when compared to non-adapted cells. The results were obtained by subjecting in vitro irradiated whole blood from 10 healthy volunteers (including 2 A-bomb survivors carrying 1.5-2 Gy in vivo exposure) in an unstimulated condition (G0) using the comet assay. The intensity of DNA damage was assessed by computing the 'tail moment'. Adaptive response (AR) was noticed in only donor 3, as indicated by reduced tail moment when the blood samples received priming + challenging doses over a 4 h interval. The priming dose was either 0.01 Gy 137Cs gamma-rays or 0.0025 Gy 252Cf neutrons. The delivered challenging dose was either 1 Gy 60Co g-rays or 0.25 Gy 252Cf neutrons. The irradiation was conducted using the HIRRAC facility. A prior exposure to 0.0025 Gy 252Cf neutrons nullified the excess tail moment caused by 0.25 Gy neutrons given during a 4 h gap. In a similar way, 0.01 Gy 137Cs gamma-rays offered a cross-adaptive response to the neutron challenging dose. The tail moment of A bomb survivors after in vitro irradiation was less than that of the age-matched control and, at the same time, was not influenced by the priming dose. An altered subset and the immunological status of blood after A-bomb exposure were cited as possible factors. Because AR can affect the outcome of RBE, its individual variability only emphasizes the need to have individual biodosimetry for better risk assessment, especially in planning for a long space voyage. PMID- 11393894 TI - The analysis of repeated measures designs: a review. AB - Repeated measures ANOVA can refer to many different types of analysis. Specifically, this vague term can refer to conventional tests of significance, one of three univariate solutions with adjusted degrees of freedom, two different types of multivariate statistic, or approaches that combine univariate and multivariate tests. Accordingly, it is argued that, by only reporting probability values and referring to statistical analyses as repeated measures ANOVA, authors convey neither the type of analysis that was used nor the validity of the reported probability value, since each of these approaches has its own strengths and weaknesses. The various approaches are presented with a discussion of their strengths and weaknesses, and recommendations are made regarding the 'best' choice of analysis. Additional topics discussed include analyses for missing data and tests of linear contrasts. PMID- 11393895 TI - Modelling dissimilarity: generalizing ultrametric and additive tree representations. AB - Methods for the hierarchical clustering of an object set produce a sequence of nested partitions such that object classes within each successive partition are constructed from the union of object classes present at the previous level. Any such sequence of nested partitions can in turn be characterized by an ultrametric. An approach to generalizing an (ultrametric) representation is proposed in which the nested character of the partition sequence is relaxed and replaced by the weaker requirement that the classes within each partition contain objects consecutive with respect to a fixed ordering of the objects. A method for fitting such a structure to a given proximity matrix is discussed, along with several alternative strategies for graphical representation. Using this same ultrametric extension, additive tree representations can also be generalized by replacing the ultrametric component in the decomposition of an additive tree (into an ultrametric and a centroid metric). A common numerical illustration is developed and maintained throughout the paper. PMID- 11393896 TI - A computationally efficient method for obtaining standard error estimates for the promax and related solutions. AB - A computationally efficient algorithm for computing the asymptotic standard errors for the promax factor solution is proposed. The algorithm covers promax rotation with or without row normalization of the pre-rotated factor matrix. It also covers situations with either even- or odd-powered promax targets. With some modifications, the algorithm applies to Procrustean rotations with fixed or random independent targets. Simulation results show that the algorithm provides reasonable approximate standard errors for the promax solution with N = 200. In a real data example, the numerical results of the standard error computation using the proposed algorithm match those of an existing method based largely on the augmented information approach. The reasons why the proposed algorithm is more efficient than the augmented information approach are discussed. PMID- 11393897 TI - A minimax procedure in the context of sequential testing problems in psychodiagnostics. AB - The purpose of this paper is to derive optimal rules for sequential testing problems in psychodiagnostics. In sequential psychodiagnostic testing, each time a patient is exposed to a new treatment, the decision then is to declare this new treatment effective, ineffective, or to continue testing and exposing the new treatment to another random patient suffering from the same mental health problem. The framework of minimax sequential decision theory is proposed for solving such testing problems; that is, optimal rules are obtained by minimizing the maximum expected losses associated with all possible decision rules at each stage of testing. The main advantage of this approach is that costs of testing can be explicitly taken into account. The sequential testing procedure is applied to an empirical example for determining the effectiveness of a cognitive-analytic therapy for patients suffering from anorexia nervosa. For a given maximum number of patients to be tested, the appropriate action is indicated at each stage of testing for different numbers of positive reactions to the cognitive-analytic therapy. The paper concludes with a simulation study, in which the minimax sequential strategy is compared for the anorexia nervosa example with other procedures that exist for similar classification decision problems in the literature in terms of average number of patients to be tested, classification accuracy and average loss. PMID- 11393898 TI - Effect of outliers on estimators and tests in covariance structure analysis. AB - A small proportion of outliers can distort the results based on classical procedures in covariance structure analysis. We look at the quantitative effect of outliers on estimators and test statistics based on normal theory maximum likelihood and the asymptotically distribution-free procedures. Even if a proposed structure is correct for the majority of the data in a sample, a small proportion of outliers leads to biased estimators and significant test statistics. An especially unfortunate consequence is that the power to reject a model can be made arbitrarily--but misleadingly--large by inclusion of outliers in an analysis. PMID- 11393899 TI - Conditional local influence in case-weights linear regression. AB - The local influence approach proposed by Cook (1986) makes use of the normal curvature and the direction achieving the maximum curvature to assess the local influence of minor perturbation of statistical models. When the approach is applied to the linear regression model, the result provides information concerning the data structure different from that contributed by Cook's distance. One of the main advantages of the local influence approach is its ability to handle the simultaneous effect of several cases, namely, the ability to address the problem of 'masking'. However, Lawrance (1995) points out that there are two notions of 'masking' effects, the joint influence and the conditional influence, which are distinct in nature. The normal curvature and the direction of maximum curvature are capable of addressing effects under the category of joint influences but not conditional influences. We construct a new measure to define and detect conditional local influences and use the linear regression model for illustration. Several reported data sets are used to demonstrate that new information can be revealed by this proposed measure. PMID- 11393900 TI - Some Mantel-Haenszel tests of Rasch model assumptions. AB - A class of Rasch model tests is proposed, all of them based on the Mantel Haenszel chi-squared statistic. All tests make use of the 'sufficient statistics' property the Rasch model possesses. One element of our general class, the test for item bias developed by Holland and Thayer, has been discussed extensively in the psychometric literature. Three applications of the general procedure are presented, two on unidimensionality and one on item dependence in educational testing. In each case, simulation results are reported. Our procedure is also applied to real data. PMID- 11393901 TI - Inferences about correlations when there is heteroscedasticity. AB - Let (Yb Xi), i = 1, ..., n, be a random sample from some bivariate distribution, and let rho be the (Pearson) population correlation between X and Y. The usual Student's t test of H0: rho = 0 is valid when X and Y are independent, so in particular the conditional variance of Y, given X, does not vary with X. But when the conditional variance does vary with X, Student's t uses an incorrect estimate of the standard error. In effect, when rejecting H0, this might be due to rho not equal to 0, but perhaps the main reason for rejecting is that there is heteroscedasticity. This note compares two heteroscedastic methods for testing H0 and finds that in terms of Type I errors, the nested bootstrap performed best in simulations when using rho. When using one of two robust analogues of rho (Spearman's rho and the percentage bend correlation), little or no advantage was found, in terms of Type I error probabilities, when using a nested bootstrap versus the basic percentile method. As for power, generally an adjusted percentile bootstrap, used in conjunction with r, performed better than the nested bootstrap, even in situations where, for the null case, the estimated probability of a Type I error was lower when using the adjusted percentile method. As for computing a confidence interval when correlations are positive, situations are found where all methods perform in an unsatisfactory manner. PMID- 11393902 TI - Three-mode models for multivariate longitudinal data. AB - Multivariate longitudinal data are characterized by three modes: variables, occasions and subjects. Three-mode models are described as special cases of a linear latent variable model. The assumption of measurement invariance across occasions yields three-mode models that are suited for the analysis of multivariate longitudinal data. These so-called longitudinal three-mode models include autoregressive models and latent curve models as special cases. Empirical data from the field of industrial psychology are used in an example of how to test substantive hypotheses with the longitudinal, autoregressive and latent curve three-mode models. PMID- 11393903 TI - Using Johnson's transformation and robust estimators with heteroscedastic test statistics: an examination of the effects of non-normality and heterogeneity in the non-orthogonal two-way ANOVA design. AB - The present study proposes a procedure that combines Johnson's transformation and the trimmed means method to deal with the problem of non-normality. An approximate test such as the Alexander-Govern test or Welch-James type test is then employed to deal with the heterogeneity of cell variance in the non orthogonal two-way fixed effects completely randomized design. Both unweighted and weighted means analyses are considered. The empirical Type I error rates and the statistical power for comparing population means are investigated by Monte Carlo simulation. The simulated results show that Johnson's transformation with trimmed mean and the approximate test is valid in terms of Type I error rate control, and that the magnitude of the statistical power for non-normal distributions is better than that of conventional methods. PMID- 11393904 TI - A note on covariance structure analysis methods for testing invariance of reliability and stability in multi-wave, multi-indicator models. AB - Covariance structure analysis methods for testing invariance in reliability and stability coefficients in multi-wave, multi-indicator models are discussed. Alternative approaches are outlined, which are straightforward and simpler than the reparameterization recently proposed by Alanen, Leskinen and Kuusinen. PMID- 11393905 TI - Congenital malformation: abnormal position of the malleus. PMID- 11393906 TI - Endoscopic view of a concha bullosa of the middle turbinate. PMID- 11393907 TI - Laryngeal aspergillosis. PMID- 11393908 TI - Branchial cleft cyst. PMID- 11393909 TI - Electronystagmography: patient with positional and neck torsion nystagmus. PMID- 11393910 TI - Nasopharyngeal abscess and facial paralysis as complications of petrous apicitis: a case report. AB - An 83-year-old man with a lifelong history of intermittent otorrhea and hearing loss was referred for management of a facial paralysis of 3 weeks' duration. Computed tomography (CT) of the head detected a neoplasm of the nasopharynx along with chronic otomastoiditis. A followup CT suggested the development of a nasopharyngeal abscess, which was confirmed by needle aspiration. A later coronal projection CT showed definite bone destruction in the anterior petrous apex, confirming suspicions that a petrous apicitis was responsible for the facial paralysis and abscess. This article describes the management of this patient and reviews the historical, medical, and surgical aspects of petrous apicitis. PMID- 11393911 TI - The safety and effectiveness of the Le Fort I approach to removing central skull base lesions. AB - The difficulty of gaining access to the central skull base has led to the development of many surgical approaches to this area during the past decade. Yet we believe that the Le Fort I technique, which has been used for almost 140 years in orthognathic surgery, is still an excellent approach to treating anterior skull base lesions. This procedure, which entails the horizontal sectioning of the dentoalveolar maxillary segment, seemed to fall out of favor with otolaryngologists after a few reports of complications surfaced during the past 10 to 15 years. In this article, we report a series of seven patients whom we treated with a Le Fort I approach during a 3-year period for a variety of benign and malignant anterior skull base lesions. We have encountered no significant complications of surgery or recurrence of disease at a maximum postoperative followup of 3 years. PMID- 11393912 TI - Hereditary angioedema: report of a case. AB - Hereditary angioedema is caused by an absolute deficiency or the functional inactivity of C1 esterase inhibitor in plasma. A precise diagnosis is important because, unlike allergic forms of mucocutaneous edema, this condition does not respond to epinephrine, antihistamines, or corticosteroids. We report the case of a 24-year-old man who experienced an acute attack after he had stopped taking his prophylactic medication. PMID- 11393913 TI - Structural and ultrastructural study of the anterior portion of the nasal septum and inferior nasal concha. AB - We used scanning electron microscopy to study and compare the histologic and ultrastructural aspects of the mucosa of the anterior portion of the septum (APS) and the anterior portion of the inferior concha (APINC) in 10 healthy adults and 10 cadavers. We found that (1) in most cases, the types of epithelium were generally the same in both areas--pseudostratified cylindrical ciliated epithelium with goblet cells (respiratory epithelium), stratified cuboidal epithelium, and stratified squamous nonkeratinized epithelium; (2) the APINC had more respiratory epithelium than the APS; (3) the APS had more squamous epithelium than the APINC; (4) the basement membrane of the APINC was thicker than that of the APS; (5) moderate chronic inflammatory infiltrate was more common in the APINC, and mild infiltrate was more common in the APS; and (6) the APINC had more mucous glands and the APS had more serous glands. PMID- 11393914 TI - Diagnosis and treatment of lipomas of the internal auditory canal. AB - A Chinese woman came to the otolaryngology department with a complaint of a longstanding progressive left hearing loss and intractable tinnitus. A finding of asymmetry on sensorineural hearing loss and auditory brainstem response testing provided evidence of a retrocochlear lesion on the left side. Computed tomography and magnetic resonance imaging detected a tumor in the left internal auditory canal (IAC). In addition to these radiologic features, our suspicion of the possibility of an IAC lipoma was raised by the observation of multiple lipomas over the patient's trunk and limbs. The patient underwent a complete tumor resection via the translabyrinthine approach, Pathology confirmed the diagnosis of an IAC lipoma. Although we were not able to preserve the hearing in her left ear, the patient was satisfied that we had relieved her constant tinnitus. In this article, we review the particulars of this case and discuss the clinical, radiologic, and pathologic features of IAC lipomas. PMID- 11393915 TI - Advancing efforts to address youth violence involvement. AB - Discusses the increased public attention on violence-related problems among youth and the concomitant increased diversity in research. Youth violence involvement is a complex construct that includes violence experienced in multiple settings (home, school, neighborhood) and in multiple forms (as victims, witnesses, perpetrators, and through family members, friends, and the media). Potential impacts of such violence involvement are considerable, including increased internalizing and externalizing behaviors among youth and future problems in school adjustment and life-course development. This introductory article reviews key dimensions of youth-related violence, describes an American Psychological Association Task Force (Division 12) developed to advance relevant research, and presents examples of national resources and efforts that attempt to address this critical public health issue. PMID- 11393916 TI - Assessing the status of research on violence-related problems among youth. AB - Presents results of a systematic review of abstracts on studies related to violence and youth in an effort to identify areas that have received little attention in the psychological literature and to present recommendations for future research. A total of 1,168 empirical articles on violence-related problems in youth were identified by a PsycINFO (American Psychological Association, 1980 1999) search. These articles were then classified in a multidimensional grid, allowing for comparisons among different types of articles. A review of abstracts from these articles indicated that most of the research activity has been descriptive (e.g., reviewing correlates or predictors of violence involvement) or assessment related (e.g., evaluating specific measures). Fewer articles examined the treatment or prevention of violence-related problems among youth. Further, the majority of studies pertained to direct exposure to violence (as a victim or perpetrator), with very few studies looking at the effects of witnessing violence, knowing individuals exposed to violence, or being exposed to violence through the media. Comparing treatment and prevention articles, we found that the least empirical attention was paid to the prevention of violence-related problems in youth, and not a single study was identified through this search that sought to examine the prevention of youth witnessing violence. Implications for future research agendas are discussed. PMID- 11393917 TI - A qualitative investigation of perceptions of violence risk factors in low-income African American children. AB - Conducted a qualitative investigation to identify the perceptions of risk factors for violence in a sample of inner-city African American youth. Using ethnographic analyses, themes emerging from these data included concerns about the reciprocity between drugs and violence, familial quality of life issues, gender differences in the experience of violence and risk for violence, community safety concerns, and fears about managing peer relationships specific to violence. These data are interpreted relative to the risk factors the violence prevention literature has identified among youth residing in urban environments. Findings are discussed in terms of their potential contribution to generating hypotheses for the development of theory and effective violence prevention practice. PMID- 11393918 TI - Longitudinal family and peer group effects on violence and nonviolent delinquency. AB - Explored the longitudinal relations between family relationships and parenting characteristics, violence and nonviolent delinquency of peers, and individual delinquency and violence using data from a sample of 246 adolescent male participants in the Chicago Youth Development Study. Family and parenting characteristics were measured when participants averaged 12 years of age, peer group offending when participants averaged 14 years of age, and individual offending when participants averaged 17 years of age. Family characteristics and parenting were represented by an ordinal variable ranging from exceptional families characterized by emotional closeness, strong beliefs about family, and good parenting skills, to struggling families characterized by a lack of emotional cohesion, deviant beliefs, and poor parenting. Peers' violence but not nonviolent delinquency predicted individual violence and nonviolent delinquency, and family types predicted peer deviance as well as individual violence and delinquency. Effects varied somewhat due to gang membership and ethnicity. PMID- 11393919 TI - Predictors of violence exposure among inner-city youth. AB - Examined the ability of demographic variables and risk factors (parental substance use, number of people in the home, out-of-home placements, grades repeated, arrest history, and total life stress) to predict exposure to community violence among 342 high school students from inner-city Baltimore referred for mental health care in community centers or in the schools. Over 90% of the sample knew at least 1 victim of a violent act, 77% reported witnessing a violent act, and 47% reported past victimization by violence. Risk variables were more powerful regression predictors of violence exposure than demographic characteristics such as race, sex, or clinical setting. Even after controlling for demographic differences in violence exposure, risk factors as a group accounted for another 10% to 15% of variance. Life stress was the most consistent predictor of violence exposure for this sample, and life stress was the only variable to make a significant unique contribution to the prediction of all 4 violence criteria. PMID- 11393920 TI - Emotional and behavioral impact of exposure to community violence in inner-city adolescents. AB - Used multiple methods and measures (i.e., youth report, psychiatric interviews, psychophysiological assessment) to investigate the emotional and behavioral impacts of exposure to community violence. Participants were 185 inner-city high school students (M age = 15.4 years; 42% female; 90% African American). Youth with high levels of community violence exposure reported more fears, anxiety, internalizing behavior, and negative life experiences than those with low exposure. No depression or externalizing behavior differences were observed. In a psychophysiological assessment in which adolescents watched a montage of media violence, youth exposed to high levels of community violence had lower baseline heart rates than those with low exposure. There were no between-group differences in physiologic reactivity. Regression analyses revealed that community violence exposure predicted posttraumatic stress and separation anxiety symptoms. The results suggest a significant link between community violence exposure and anxiety symptomatology. Clinical implications are discussed. PMID- 11393921 TI - Development and evaluation of school-based violence prevention programs. AB - Discusses key issues in developing and evaluating school-based violence prevention interventions. Schools provide a natural setting for implementing programs directed at teaching youth attitudes, knowledge, and skills to reduce their involvement in violence. Although multitudes of these programs exist, few have been rigorously evaluated. Developers of violence prevention programs need to pay particular attention to the type of violence being addressed, the target population, relevant risk and protective factors, and the target of the intervention. Conducting sound evaluations of such programs requires careful attention to the unit of randomization, treatment conditions, outcome measures, timing of data collection, and potential moderator variables. Efforts to develop effective prevention programs can be greatly facilitated by adopting an action research strategy in which evaluation findings provide a basis for continual program refinement. PMID- 11393922 TI - Implementation of an empirically based drug and violence prevention and intervention program in public school settings. AB - Describes the implementation of a collaborative preventive intervention project (Healthy Schools) designed to reduce levels of bullying and related antisocial behaviors in children attending two urban middle schools serving primarily African American students. These schools have high rates of juvenile violence, as reflected by suspensions and expulsions for behavioral problems. Using a quasi experimental design, empirically based drug and violence prevention programs, Bullying Prevention and Project ALERT, are being implemented at each middle school. In addition, an intensive evidence-based intervention, multisystemic therapy, is being used to target students at high risk of expulsion and court referral. Hence, the proposed project integrates both universal approaches to prevention and a model that focuses on indicated cases. Targeted outcomes, by which the effectiveness of this comprehensive school-based program will be measured, are reduced youth violence, reduced drug use, and improved psychosocial functioning of participating youth. PMID- 11393923 TI - Emerging themes and challenges in understanding youth violence involvement. AB - Identifies emerging key themes in the articles in this special section regarding child and adolescent involvement in violence. Using a developmental-ecological framework and drawing on consistencies across other studies and reviews, this article is intended to summarize the pertinent findings for clinical child psychology. Current knowledge status, needed further research, clinical implications (preventive and treatment), and relevant policy issues are noted. PMID- 11393924 TI - Children's responses to overt and relational aggression. AB - Investigated children's responses for coping with overt and relational aggression. Children in Grades 3 through 6 (N = 491) in a rural Midwestern public school district completed a survey designed to assess how students cope when they are the targets of peer aggression. Children endorsed greater use of internalizing and distancing strategies for coping with relational aggression and greater use of externalizing strategies for coping with overt aggression. In addition, older children reported greater use of externalizing and less use of internalizing and distancing strategies than younger children. Significant differences were also found between boys and girls. Regardless of type of aggression, girls endorsed greater use of problem-solving and support strategies and less use of externalizing strategies than boys. Coping of high target children and of children who frequently received prosocial treatment from peers were also examined. PMID- 11393925 TI - Revisiting the accuracy hypothesis in families of young children with conduct problems. AB - Revisited the accuracy hypothesis in an examination of the relation between maternal depressive symptomatology and child conduct problems. All data were gathered as part of the pretreatment assessment in an outcome study of families with clinic-referred children with conduct problems (age 3 to 6). The mothers varied in their depressive symptomatology, from not at all symptomatic to severely symptomatic. Correlations indicated that with increasing depressive symptomatology, mothers (N = 97) displayed a higher rate of physical negative behaviors towards their child and reported more child conduct problems. Regression analyses revealed that at the lowest levels of maternal depressive symptomatology there was a discrepancy between mothers' reports of child behavior problems and child deviant behaviors observed during mother-child interaction. In contrast, at higher levels of depression, mothers' reports of child behavior were consistent with laboratory observations of their child's behavior. These findings provide evidence to support the accuracy hypothesis in reference to mothers who display a high degree of depressive symptomatology, but the results also call into question the validity of maternal report in families with children with conduct problems. PMID- 11393926 TI - Clinical significance and correlates of disruptive behavior in environmentally at risk preschoolers. AB - Examines the clinical significance and correlates of disruptive behavior disorder symptoms (DBDSX) in preschoolers. Participants were 129 predominantly minority preschoolers (2 1/2 to 5 1/2) residing in low-income environments, half of whom were clinic-referred for disruptive behavior. Children with higher levels of DBDSX were more impaired in parent-child, preschool and clinic contexts. Correlates of DBDSX included both prenatal and infancy risks (low soothability as infants, prenatal exposure to cigarettes) and concurrent parenting factors (harshness, low levels of behavioral responsiveness, and parenting stress). In general, the clinical and risk profile of DBDSX in preschoolers at environmental risk appears to be similar to that of older children. Based on the results of this study, etiologic and prevention research on disruptive behavior disorders should begin in the first few years of life. PMID- 11393927 TI - The health and education leadership project: a school initiative for children and adolescents with chronic health conditions. AB - An estimated 40% of children and adolescents with chronic health conditions such as cancer, diabetes, and hemophilia, experience school-related problems. Many children living with a chronic illness do not need or may not qualify for traditional special education programs, yet they may have unmet learning needs associated with chronic absenteeism, disease symptoms, or side effects of medical treatments. Using principles of family-centered schools, a coalition of schools, families, pediatricians, and nurses can work cooperatively to help students with chronic conditions succeed. This article describes the second phase of an ongoing initiative designed to support public schools in their efforts to meet the health and education needs of students with chronic health conditions. PMID- 11393928 TI - School-based telehealth: an empirical analysis of teacher, nurse, and administrator perceptions. AB - In 1997, the University of Kansas Medical Center (KUMC) and the United School District 500 (USD 500) in Kansas City, Kansas, launched an innovative telehealth project that brought health care directly into elementary schools by interactive video technology and peripheral devices. This paper focuses on the attitudes and opinions of key players--teachers, nurses, and administrators--involved in providing the service. Research methodology involved analyzing archival data and conducting interviews with key players on the project from USD 500 and KUMC. The study highlights the difficulty in delivering health care, especially telehealth care, to underserved, urban children. The data also reveal that initial challenges and negative attitudes can be overcome to effectively deliver telehealth care in the school setting. PMID- 11393929 TI - Safety makes sense: a program to prevent unintentional injuries in New York City public schools. AB - Estimates suggesting that 10% to 25% of unintentional injuries to school-aged children occur in and around American schools are alarming. Information from the National Health Information Survey indicates as many as 3.7 million children are injured in schools annually. To address these concerns, The New York Academy of Medicine and the Board of Education of the City of New York in 1998 launched Safety Makes Sense, a multidimensional school-based, injury prevention initiative. The Safety Makes Sense program integrates unintentional injury prevention into comprehensive school health education in elementary and middle school classrooms and is implemented by interdisciplinary teams within a coordinated school health program. The multifaceted program also integrates safety and music education, and includes development of a handbook for teachers and staff, student booklets, staff development training workshops, and evaluation. PMID- 11393930 TI - Factors associated with student participation in a school-based hepatitis B immunization program. AB - This study examined the relationship between participation in a school-based hepatitis B immunization program and teacher attitudes toward school-based health care and student socioeconomic factors. A survey addressing teachers' attitudes was administered to all teachers participating in the program. Information regarding student participation in school lunch programs and scores on national standardized tests were collected. Of the 4,874 fifth-grade students targeted for the program, 3,483 (72%) consented to be vaccinated and 3,232 (93% of 3,483) received all three doses of vaccine. Socioeconomic factors were the most important predictors of student participation in this school-based immunization program. Participation was significantly lower among students in schools with a high proportion of students receiving free or reduced-price school lunch and with low test scores. The only teacher factor associated with student participation was whether the teacher had returned the questionnaire. Strategies to increase immunization coverage in school-based programs should target children of low socioeconomic status. PMID- 11393931 TI - Effectiveness of the "Baby Think It Over" teen pregnancy prevention program. AB - In an effort to reduce teen pregnancy, schools are purchasing a program called "Baby Think It Over," a computerized infant simulator intended to provide a realistic infant care experience. However, little empirical, especially experimental, program effectiveness data exist. This study determined if the program changed participants' attitudes toward parenting, as well as sexual and contraceptive behaviors linked to avoidance of teen pregnancy. Development of measurement tools was a part of the purpose. The study also asked teen-agers, through narrative questions, about their conscious perceptions of the baby's utility and impact. Participants included experimental (n = 151) and control (n = 62) groups of primarily White, middle class, suburban high school' students (mean age = 16.2). The quasi-experimental portion of the study failed to reveal a statistically significant effect. Narrative data revealed several positive and notable program effects. PMID- 11393932 TI - Impact of a school-based health center on emergency department use by elementary school students. AB - While school-based health centers (SBHC) improve student access to health services, it remains unclear if use of the centers can reduce hospital emergency department visits. This study evaluated the impact of an elementary school SBHC on emergency department visits by children enrolled in the center. Major reasons for visits included trauma (32%), otitis media (15%), upper respiratory infections (9%), and gastroenteritis (6%). Implementation of an elementary SBHC resulted in a significant decrease (p < 0.03) in non-urgent emergency department visits. No difference existed in urgent emergency department visits. Medicaid insured children were more likely to use the emergency department than privately insured or uninsured children. Reducing emergency department visits can decrease medical costs and support the cost effectiveness of SBHCs. PMID- 11393933 TI - Utilizing undergraduate nursing students to provide health education in elementary schools. PMID- 11393934 TI - Contextual control of appetitive conditioning: influence of a contextual stimulus generated by a partial reinforcement procedure. AB - Three experiments with rat subjects examined the effects of a context switch after conditioning treatments in which the conditioned stimulus (CS) was either paired with food on every presentation (continuous reinforcement) or on some of its presentations (partial reinforcement). In each experiment, a target CS was given one of these treatments in Context A, and another CS was given a treatment during sessions that were intermixed in Context B. Final tests of the target CS in Context A and Context B often revealed no loss of responding with the switch to B. However, a loss was observed when partial reinforcement had been associated with Context A and continuous reinforcement had been associated with Context B. Those conditions caused equal decrements in responding to partially reinforced and continuously reinforced targets. The results suggest that under the present conditions partial reinforcement can generate a contextual stimulus that becomes associated with the physical context and controls responding to the CS. PMID- 11393935 TI - The role of nonreinforcement in the learning of honeybees. AB - Two series of experiments with honeybees were designed to test the assumption that inhibition is generated by nonreinforcement as a function of the excitatory value of the context. In the first series (Experiments 1-3), summation tests with B were made after A+/C-/AB- as compared to A+/C-/CB- training, with precautions taken to minimize the possibility of a masking effect of excitatory within compound conditioning on AB trials; responding to B did not vary with training procedure. In the second series (Experiments 4-5), retardation tests rather than summation tests were used, in the belief that they might be more sensitive; after A+/AB-/CD- training, acquisition in a B+/D- problem was found to be no less rapid than in a D+/B- problem. A third series of experiments (Experiments 6-9) was designed to test the more general assumption that the effectiveness of nonreinforcement increases with the excitatory value of the context; response to B was found to be no different after A+/B+/C- training followed by A+/AB- training than after A+/B+/C- training followed by A+/CB- training. The results are compatible with the view that the role of nonreinforcement in honeybees is not to generate inhibition, but only to reduce excitation in a manner independent of the excitatory value of the context. PMID- 11393936 TI - Mediated conditioning and retrospective revaluation with LiCl then flavour pairings. AB - It is becoming accepted that the associative strength of a cue can change in its absence, despite this being difficult to explain using existing theories of Pavlovian conditioning. To investigate the influence of timing on learning about the representation of an absent cue, lithium chloride (LiCl) or a flavour previously paired with LiCl was presented in a distinctive context that had previously been paired with a neutral target flavour. The former treatment produced an aversion to the target flavour whether the LiCl was presented 10 min before, or immediately after, exposure to the context. However, presenting the flavour associate of LiCl created an aversion to the target flavour only if it had been presented 10 min after LiCl during initial training. This pattern of results cannot be explained in the terms of a simple timing account, and it is proposed that an explanation will require different associative rules operating in simultaneous and successive training schedules. PMID- 11393937 TI - Interference with judgements of control and learning as a result of prior exposure to controllable and uncontrollable feedback during concept-learning tasks. AB - Two experiments examined whether exposure to an uncontrollable relationship between an action and its outcome during a nonaversive pretreatment phase would attenuate subsequent ratings of control given to actions emitted by subjects. In Experiment 1, such an interference effect was demonstrated relative to a group that received prior training with a controllable action-outcome relationship, and relative to a group not exposed to any prior relationship. In Experiment 2, these effects were replicated, and interference was also found to occur when learning a maze task. Thus, the effects of helplessness were shown to be quite general, to be produced by a nonaversive induction procedure, and to occur most readily when the current contingency between action and outcome was weakest. PMID- 11393938 TI - Evidence for inhibitory associations between the unique elements of two compound flavours. AB - In each of two experiments, rats were pre-exposed to two flavoured solutions, saline-lemon and sucrose-lemon. For group ALT, trials with one solution alternated with trials with the other. Group BLK received all trials with one solution in a block, before any trials with the other. An associative theory suggests that the alternating, but not the blocked, schedule would establish an inhibitory association between sucrose and saline. To provide a retardation test of this inhibition, some animals in each group were then given a single pairing of saline and sucrose, experienced sodium depletion, and were finally tested for their consumption of sucrose. Sodium depletion increased consumption of sucrose more in group BLK than in group ALT. In groups given no saline-sucrose pairing, sodium depletion had only a small effect on sucrose consumption, which was the same in both groups. After multiple pairings of saline and sucrose, sodium depletion had an equally large effect on sucrose consumption in both ALT and BLK groups. These results imply that alternating pre-exposure to two compound solutions does establish an inhibitory association between their unique elements, and thus provide support for an associative theory of perceptual learning. PMID- 11393940 TI - End-position nystagmus as an indicator of ethanol intoxication. AB - The Horizontal Gaze Nystagmus test is used by law enforcement agencies in the United States to determine whether drivers are intoxicated. It has a high baseline error and a dose/response relationship that varies greatly according to whether the subject's blood alcohol concentration is rising or falling. Confusion exists among practitioners of the test about whether it quantifies alcohol concentration or evaluates impairment. Fatigue exacerbates one component of the HGN test, end-position nystagmus. Video tapes recorded by cameras in police vehicles revealed that police officers rarely comply with the minimum requirements of the nystagmus examination procedures for which they were trained and certified. PMID- 11393941 TI - Screening for canine spermatozoa. AB - A case is presented in which possible canine spermatozoa were encountered in the routine examination of sexual assault evidence. Some observations and difficulties concerning the detection of canine spermatozoa are discussed. A possible visual screening method for canine sperm cells is offered, involving conventional forensic/histological staining techniques and the use of light microscopy. PMID- 11393942 TI - Science and justice from a prosecutor's perspective. PMID- 11393943 TI - "Best value" in forensic pathology. PMID- 11393945 TI - The fallacy of independence testing and the use of the product rule. AB - Use of the product rule which implies the assumption of within and between locus independence, is still common, particularly in the United States of America. Whilst it may be considered by some to be an acceptable approximation it is not logical to suggest that independence testing somehow "validates" its use. This paper discusses the nature of this fallacy. PMID- 11393946 TI - The scientific basis for human bitemark analyses--a critical review. AB - This article presents a discussion of the scientific basis for human bitemark analyses. Using a review of the literature, the major areas of contention within the field are assessed: including the accuracy of bitemarks on skin, the uniqueness of the human dentition, and analytical techniques. The review revealed a lack of valid evidence to support many of the assumptions made by forensic dentists during bitemark comparisons. The new level of judicial scrutiny of such scientific evidence is likely to emphasise this lack of knowledge upon which bitemark analysis relies. The authors call for a more scientific and evidence based approach to forensic dental research. PMID- 11393948 TI - [The thickness change and clinical significance of tunica intima and smooth muscle in artery after hydrolic dilation]. AB - OBJECTIVE: To study the proliferation change of tunica intima and smooth muscle in artery after hydrolic dilation for potential clinical use. METHODS: Sixten adult New Zealand rabbits were randomly divided into 4 groups, named group A, B, C and D. Right carotid arteries of rabbits of those 4 groups were dilated by hydrolic dilation with different pressures with 0 kPa, 40 kPa, 80 kPa, and 120 kPa respectively. The arterial calibers, thickness of tunica intima and smooth muscle were analyzed by automatic medical photograph analyzer immediately, 1 week and 2 weeks later respectively. RESULTS: The arterial calibers in the experimental group were larger than those in control group after immediate hydrolic dilation and 1 week later (P < 0.01). At 2 weeks, the arterial calibers in group B and D has no significant difference compared to group A (P > 0.05), and those in group C were larger than that of group A (P < 0.01). There were no significant difference in thickness of tunica intima and smooth muscle between the experimental group and control group (P > 0.05) after immediate hydrolic dilation. At 1 and 2 weeks after dilation, there were no significant difference between group A and group B (P > 0.05), and those in group C and D were all larger than those in group A (P < 0.01). No obvious proliferation of tunica intima were observed in group B at 2 weeks after hydrolic dialation, but the proliferation of tunica intima could be observed in group C and D, especially in group D. CONCLUSION: Caliber of artery can be expanded by hydrolic dilation with higher pressure, but the proliferation of tunica intima and smooth muscle may be occurred in hydrolic dilation with higher pressure over 80 kPa, therefore it is safe to use hydrolic dilation with pressure no more than 40 kPa. PMID- 11393949 TI - [Clinical effects of revascularization in lower limb ischemia]. AB - OBJECTIVE: To investigate the clinical effects of revascularization in lower extremity for severe ischemia. METHODS: Fifty-six lower limbs with severe ischemia in 49 patients were evaluated retrospectively, who underwent surgical intervention from January of 1995 to December of 2000. By arteriography, the actual anatomic distributions of occlusive disease included infrarenal aorta bicommon iliac arteries, abdominal aorta-bicommon iliac arteries, iliac artery, and femoral artery or femoropopliteal artery. The indication for surgery was disabling claudication, rest pain and gangrene. Fourteen limbs in 12 cases received arterialization of femoral venous system by artificial venous-arterial fistula. Artificial vascular grafts were implanted in 33 limbs of 28 cases, endarterectomy and patch profundaplasty were performed in 5 limbs of 5 cases, and primary amputation was carried out in 4 cases. RESULTS: During 38 months follow up in average, 4 limbs were amputated within 52 revascularizated limbs, and accumulated amputation rate was 14.3%. Patency rate was 68.4% in arterial revascularization limbs (26/38 limbs), and limb survival rate was 94.7%(36/38 limbs) by procedure of artificial vascular grafts, endarterectomy and patch profundaplasty. Limb survival rate in procedure of artificial venous-arterial fistula was 85.7%(12/14 limbs). CONCLUSION: In treatment of severe lower extremity ischemia, the effective revascularization can be achieved by artificial vascular bypass, endarterectomy and patch profundaplasty, or arterialization of femoral venous system. Options in the surgical management should depend on individual. Arteriography is essential for revascularization and properly planning a practicable surgical approach. PMID- 11393950 TI - [Anatomical study of reconstruction of vertebral artery with neighboring non trunk arteries]. AB - OBJECTIVE: To study the anatomical basis for reconstruction of vertebral artery with neighboring non-trunk arteries. METHODS: Twenty preserved adult cadavers were used in this study to observe the morphology of superior thyroid artery, inferior thyroid artery, transverse cervical artery, thyrocervical trunk and extracerebral portion of vertebral artery, and reconstruction of vertebral artery with these arteries was simulated in two preserved cadavers. RESULTS: The calibers of superior or inferior thyroid artery, or transverse cervical artery were more than 2 mm in diameter, and the arteries had suitable free length for end-to-side anastomosis with vertebral artery. Thyrocervical artery had similar caliber to vertebral artery so that end-to-end anastomosis could be carried out between them, but only 38.5% of this artery had adequate artery trunk (more than 10 mm). It was proved from the simulated procedures that the reconstruction of vertebral artery with these neighboring non-trunk arteries was possible. CONCLUSION: Reconstruction of vertebral artery with neighboring non-trunk arteries has anatomical basis and can be used clinically for treatment of the lesion affecting the first or second portion of vertebral artery. PMID- 11393951 TI - [Experimental study on the hemostatic properties of collagen sponge]. AB - OBJECTIVE: To validate the hemostatic properties of collagen sponge made in China. METHODS: The experimental model of superficial cut of liver was established in 20 Sprague-Dawley adult rats, which were divided into two groups randomly. Collagen sponge or gelatin sponge was used to cover the cut respectively. Hemostatic result was observed. Afterwards, standard liver trauma model by resection left front liver lobe was made, wound was treated with collagen sponge or gelatin sponge respectively. Hemostatic result was observed. Concurrent hemostatic time and bleeding amount were noted. At 7, 14 and 20 days after operation, intra-abdominal adhension, infection and healing state of liver were observed by exploratory laparotomy. The histological changes of regenerate liver tissue were observed by microscopy. RESULTS: Collagen sponge adhered to wound well. Concurrent hemostatic time and bleeding amount in collagen sponge group were superior to those of gelatin sponge (P < 0.05). The histological examination showed that collagen sponge was absorbed and degraded rapidly, regenerative hepatocytes could be induced. CONCLUSION: Collagen sponge has fine hemostatic properties and can induce regeneration of hepatocytes effectively. It is worth popularizing for its convenience in clinical application and its properties of rapid degradation and absorption. PMID- 11393952 TI - [Reconstruction of nail folds by double pulp flap in congenital complete syndactyly release]. AB - OBJECTIVE: To introduce a surgical approach for reconstruction of nail folds in congenital complete syndactyly release. METHODS: A narrow flap and a broad flap were raised on the common distal phalanx to cover the denuded nail-edge in 30 fingers of 15 cases whose webs were separated. RESULTS: All of the flaps were successfully transferred and survived. The reconstructed nail folds had satisfied figure in 21 out of 30 fingers. The nail folds in the other 9 fingers, covered by a broad flap in 2 fingers and by a narrow flap in 7 fingers, were a little smaller than normal. All of the 30 fingers had normal fullness of pulp and no twisty nails. CONCLUSION: The reconstruction of nail folds by double pulp flap can be performed with a one-stage technique, and the outcome is satisfactory, which make it as a good surgical approach to reconstruct nail folds in congenital complete syndactyly release. PMID- 11393953 TI - [Clinical application of the medial multiplex flap pedicled with the posterior tibial vessel]. AB - OBJECTIVE: To investigate the clinical results of the medial multiplex flap pedicled with the posterior tibial vessel. METHODS: Twelve cases with soft tissue defects and bone defects of limbs were treated with the medial multiplex flap pedicled with the posterior tibial vessel from September 1992 to May 1999. Among them, bone and soft tissue defects following opened fracture in 7 cases, chronic ulcer following chronic osteomyelitis in 2 cases, melanoepithelioma in 2 cases, bone and soft tissue defects following osteoma resection in 1 case. The bone defect area was from 2.5 cm x 5.0 cm to 4.5 cm x 11.0 cm. Free graft was performed in 5 cases, bridged transposition in 3 cases and reversal transposition in 4 cases, among them, periosteal myocutaneous flap with autogenous or allogeneic bone grafting in 8 cases, myocutaneous flap in 4 cases. The area of the flaps from 6 cm x 8 cm to 12 cm x 25 cm. RESULTS: All flaps were healed by first intention, but in the distal fragments of bigger flaps were partially necrosed in 2 cases. In 10 cases bone healing were obtained after 16 weeks of operation according to the X-ray photos. All cases were followed up from 6 to 18 months. All cases achieved satisfactory result but 1 case died because of lung metastasis of osteoma. CONCLUSION: The multiplex graft pedicled with the posterior tibial vessel is an ideal graft for repairing the large soft tissue defects and bone defects, because it has such advantages as adequate blood supply, big vascular diameter, long pedicle and big dermatomic area. PMID- 11393954 TI - [The design of axial pattern flap with color Doppler flow imaging technique]. AB - OBJECTIVE: To study the effect of color doppler flow imaging(CDFI) technique in the design of axial pattern flap. METHODS: From April 1996 to June 1999, 10 patients with residual wound were adopted in this study. Among them, there were seven males and three females, the area of wounds ranged from 6 cm x 8 cm to 15 cm x 20 cm. Before operation, the axial pattern flaps were designed by traditional method, then CDFI technique with high frequency(5.0-7.5 MHz) was used for examining the major supply artery of the flap. At last, the modified flaps were transferred to cover the wounds. RESULTS: All the patients except one case completed the operation successfully. The cosmetic and function of the flaps were excellent. CONCLUSION: CDFI is a simple, direct and accurate method for detecting the supply artery of axial pattern flap. This technique should be popularized to avoid the blindness of flap design. PMID- 11393955 TI - [Repair of medial collateral ligament defect of knee joint with transposition of great adductor muscular tendon pedicled vessels]. AB - OBJECTIVE: To study the effect of transposition of great adductor muscular tendon pedicled vessels in repairing the medial collateral ligament defect of knee joint. METHODS: From September 1991 to September 1999, on the basis study of applied anatomy, 30 patients with the medial collateral ligament defect were repaired with great adductor muscular tendon transposition pedicled vessels. Among them, there were 28 males and 2 females, aged 26 years in average. RESULTS: Followed up for 17 to 60 months, 93.3% patients reached excellent or good grades. No case fell into the poor grade. CONCLUSION: Because the great adductor muscular tendon is adjacent to the knee joint and similar to the knee ligament, it is appropriate to repair knee ligament. Transposition of the great adductor muscular tendon pedicled vessels is effective in the reconstruction of the medial collateral ligament defect of knee joint. PMID- 11393956 TI - [Construction of human bone morphogenetic protein-2 expressing eukaryotic vector]. AB - OBJECTIVE: To construct human bone morphogenetic protein-2(hBMP-2) expressing eukaryotic vector and observe whether it can be expressed in eukaryotic cells. METHODS: pUC19 was digested with Sal I and Xba I. The resulting Sal I-Xba I fragment (1.24 kb) which contains the full length of human BMP-2 cDNA was separated on agarose gel and ligated into eukaryotic expression vector pcDNA3 digested with XhoI and XbaI. The recombinant pcDNA3-hBMP-2 plasmid was transferred into fibroblasts cell line NIH3T3. The stable expression of hBMP-2 in the positive cells G418 selected was determined by in situ hybridization and immunohistochemical analysis. RESULTS: The two fragments digested from recombinant pcDNA3-hBMP-2 plasmid by EcoR I and Xba I represented 1.3 kb and 5.38 kb respectively by agarose electrophoresis, meanwhile the Xho I site was disappeared in pcDNA3-hBMP-2 indicating the successful construction of recombinant pcDNA3-hBMP-2 plasmid. Stable expression of hBMP-2 in pcDNA3-hBMP-2 transfected cells was confirmed by in situ hybridization and immunohistochemical analysis. CONCLUSION: hBMP-2 expressing eukaryotic vector is successfully constructed and can be expressed in eukaryonic cells. PMID- 11393957 TI - [The influence of hyaluronic acid and basic fibroblast growth factor on the proliferation of ligamentous cells]. AB - OBJECTIVE: To observe the effects of hyaluronic acid (HA) and basic fibroblast growth factor (bFGF) on the proliferation of the cells from medial collateral ligament (MCL) and anterior cruciate ligament (ACL) cells. METHODS: The MCL cells and ACL cells of mature New Zealand white rabbit were cultured, while HA, bFGF or HA and bFGF were added to the cell culture media, the cellular proliferation was assayed by MTT method. RESULTS: HA only had no effect on the preoliferation of ACL cells, but had a small stimulatory effect on the proliferation of MCL cells. The addition of 1 ng/ml bFGF enhanced the proliferation of both MCL and ACL cells significantly, and this enhancement was maximal in the concentration of 50 ng/ml. However, the enhancement of proliferation of MCL and ACL cells could be achieved when the combination of HA in concentration of 100 micrograms/ml and bFGF in concentration of 100 ng/ml. CONCLUSION: It is evident that bFGF can enhance the proliferation of the ligament cells. HA can maintain the normal growth of ACL cells with no effect on the proliferation of the cells, while HA has a small stimulatory effect on the proliferation of MCL cells. However, when bFGF is coordinated with HA, more improvement of cellular proliferation can be achieved. HA can be used as a potential carrier for bFGF to enhance the healing of ligamentous tissue injuries. PMID- 11393958 TI - [Effect of improved topical agents on healing time of deep second-degree burn wound]. AB - OBJECTIVE: With the recognization of the mechanism of wound healing, some topical agents are created and applied in trauma to improve the healing rate of wounds. The main purpose of this study is to investigate the effect of some topical agents on the healing rate of deep second-degree burn wounds. METHODS: One thousand five hundred and sixty-three patients with deep second-degree burn wounds(total burn surface area < or = 10%) were involved in this study from January 1982 to December 1999. According to the application time of different treating measures including supplement of Zn, application of growth factors and collagenase, the patients were divided into 3 groups, wound healing rates were compared. RESULTS: Before 1991, none of special topical agents were used, and the healing time of deep second-degree burn wounds was(23.8 +/- 3.5) days. From 1991 to 1996, with the topical application of SD-Ag-Zn, which can provide Zn for cells taking part in wound healing, the healing time of deep second-degree burn wounds was (20.6 +/- 3.2) days, earlier than no special topical agents (P < 0.05). From 1997 to 1999, growth factors such as basic fibroblast growth factor(bFGF) and epithelial growth factor (EGF) and collagenases were applied in wound treatment combining with SD-Ag-Zn, wound healing time was (16.2 +/- 2.8) days, earlier than no special topical agents (P < 0.01) and simple SD-Ag-Zn application (P < 0.05). CONCLUSION: It indicates that the improvement of topical agents can accelerate wound healing speed. PMID- 11393959 TI - [Biliary tract reconstruction after cystectomy of congenital choledochal cyst]. AB - OBJECTIVE: To evaluate the operative methods of biliary tract reconstruction after cystectomy of congenital choledochal cyst(CCC). METHODS: One hundred and six cases with CCC underwent cystectomy and biliary tract reconstruction in our hospital from July 1984 to December 1999 were followed up. Among them, there were three kinds of procedures in biliary tract reconstruction: with single Roux-Y hepaticojejunostomy in 48 cases, with intussusceptive valve to the line of Roux-Y hepaticojejunostomy in 37 cases, with rectangular valve to the line of Roux-Y hepaticojejunostomy in 21 cases. RESULTS: Fifty nine cases were followed up for 4.68 years in average. There were 3 cases with ascending cholangitis after single Roux-Y hepaticojejunostomy. And the symptom disappeared in 2 cases underwent reoperation with an intussueceptive valve plasty to the line of Roux-Y hepaticojejunostomy. No patients suffered from ascending cholangitis in the precautionary valve plasty group. CONCLUSION: It suggests that the postoperative ascending cholangitis can be prevented effectively if standard cystectomy and prophylactic intussusceptive valve added to the line of Roux-Y hepaticojejunostomy are carried out. The procedure should be performed as soon as possible providing the child is tolerable. PMID- 11393960 TI - [Progress of treatment of wrist disorder by limited intercarpal arthrodesis]. AB - OBJECTIVE: To review the recent progress in the treatment of wrist disorder by limited intercarpal arthrodesis and the related experimental study. METHODS: Recent original articles related to limited intercarpal arthrodesis, including clinical practice and experimental study, were extensively retrieved and carefully analyzed. RESULTS: Limited intercarpal arthrodesis could relieve pain and stabilize the wrist joint with partial motion. CONCLUSION: With suitable indication and well selected operation approach, the limited intercarpal arthrodesis should be the optimal surgical intervention than total carpal athrodesis in the treatment of wrist disorder. PMID- 11393961 TI - [Proliferation properties and telomerase activity of human embryonic tendon cells transformed by ptsA58H plasmid]. AB - OBJECTIVE: To analysis the proliferation properties and telomerase activity of human embryonic tendon cells transformed by ptsA58H plasmid cultured in vitro continuously. METHODS: The 40th, 70th, and 75th passages of transformed human embryonic tendon cells (THETC) were adopted. The collagen secretion of THETC was detected by immunohistochemical methods, the growth curve of different passages of THETC was compared, and chromosome karyotype was analyzed. Total RNA of THETC were extracted to detect human telomerase reverse transcriptase (hTERT) mRNA expression by RT-PCR technique. RESULTS: When THETC were subcultured to 70 passages, the morphological characteristics of cells changed and began replicative senescence. THETC still could secret type I collagen normally. The chromosome of THETC was heteroploid (2n = 94). There were no hTERT mRNA expression. CONCLUSION: SV40 transfection can not make human embryonic tendon cells immortalization, on the other hand, human embryonic tendon cells transformed by ptsA58H plasmid has no tendency of malignant transformation. PMID- 11393962 TI - [Neocartilage of predetermined shapes]. AB - OBJECTIVE: To study chondrogenesis of calcium alginate-chondrocytes predetermined shapes. METHODS: Chondrocytes isolated from ears of rabbit by type II collagenase digestion, and then were mixed with 1.5% solidium alginate solution. The suspension was gelled to create three spatial shapes as triangle, circle and quadrilateral by immersed into 2.5% CaCl2 for 90 minutes, and then was implanted into the subcutaneous pocket on the dorsum of the rabbit. Samples were harvested at 6 and 12 weeks after implantation. RESULTS: Gross examination of excised specimens at 6 and 12 weeks after implantation revealed the presence of new cartilage of approximately the same dimensions as the original construct. Histologic evaluation using hematoxylin and eosin stains confirmed the presence of cartilage nodules at 6 weeks after implantation. After 12 weeks, mature cartilage was observed and histologic analysis confirmed the presence of well formed cartilaginous matrix. CONCLUSION: Predetermined shapes neocartilage can be regenerated using calcium alginate as a carrier of chondrocytes in the bodies of immune animals. PMID- 11393963 TI - [Expression of endogenic bone morphogenetic protein in repairing rabbit skull with tissue engineering technique]. AB - OBJECTIVE: To explore the distribution and effect of endogenic bone morphogenetic protein (BMP) in repairing rabbit skull with tissue engineered bone. METHODS: The autologous osteoblast-like cells were instantly implanted onto polyglycolic acid (PGA) matrix coated with collagen. The rabbit skull defect models were established by resection of bilateral 1.5 cm x 1.0 cm full-thickness parietal bone in 18 New Zealand rabbits, which were randomly divided into two groups. In one group, the composite of osteoblast- like cells and PGA matrix were grafted into the defect on one side of the skull as experimental group I, leaving the same defect area on the other side as control group without any graft implanted. In the other group, simply PGA was done in the same way as experimental group II. The tissue samples were harvested at 3, 8 and 14 days postoperatively and examined by histological and immunohistochemistry methods. The concentrations of BMP in different regions of the samples were measured using computer image analysis system. RESULTS: After 3 days of operation, the BMP positive cells were found in the matrix of experimental group I. At 8 days postoperatively, the formation of new bone on experimental group I was prior to that of experimental group II and control group. On the 14th day, bone trabecula was formed on the experimental group I, but there was only fibrous tissue on control group. The concentration of BMP on the experimental group I and II were higher than that of corresponding region on control side. CONCLUSION: The osteoblast-like cells instantly implanted onto PGA matrix can synthesize and secrete BMP. It may be one of the reasons of tissue engineered bone inducing new bone regeneration that localizing endogenic BMP in bone defect area, increasing the concentration of endogenic BMP and improving its distribution by tissue engineering technique. PMID- 11393964 TI - [Experimental study of bone repair induced by cryopreserved allograft periosteum and fetal bone composition in bone defect]. AB - OBJECTIVE: To investigate the repairing effect of transplantation of allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft to bone defect. METHODS: Twenty Long-eared white male rabbits were chosen as experimental model of bilateral 12 mm combined bony and periosteal radial defect. Cryopreserved allograft periosteum with allogeneic fetal bone were implanted in the left defect as experimental side and fetal bone was simply transplanted in the right defect as control side. Bone repair process in the two groups were compared by macroscopy, microscopy, roentgenograms and the contents of calcium and phosphate in the defect area at 2, 4, 8 and 12 weeks after transplantation. RESULTS: There was significant statistic difference in the contents of calcium and phosphate between the experimental and control sides at 4, 8 and 12 weeks after transplantation (P < 0.05). With time passing by, the contents of calcium and phosphate have the increasing trends. In the experimental group, lamella bone was seen and medullary canal recanalized at 8 weeks postoperatively. The histological section showed the bone lacuna and lamella bone were formed. CONCLUSION: It suggests that allogeneic fetal bone in combination with a covering cryopreserved periosteal allograft can promote bone repair, and allogeneic fetal bone is excellent bone substitute. PMID- 11393965 TI - [Trace elements and extracellular matrix]. AB - OBJECTIVE: To investigate the effects of trace elements on the metabolism of extracellular matrix and explore the physiological and pathological mechanism of trauma. METHODS: Based on the experimental and clinical data, it was studied that the action of trace elements in the metabolism of extracellular matrix in trauma repairing. RESULTS: During wound healing, the trace elements were the components of many kinds of enzymes, carriers and proteins. They took part in the synthesis of hormones and vitamins as well as the transmission of information system. They activated many different kinds of enzymes and regulate the levels of free radicals. The trace elements had the complicated effects on the synthesis, decompose, deposition and reconstruction of collagen and other extracellular matrix. CONCLUSION: The trace elements play an important role in regulating the metabolism of extracellular matrix. PMID- 11393966 TI - Environmental implications on the oxygenation of gasoline with ethanol in the metropolitan area of Mexico City. AB - Motor vehicle emission tests were performed on 12 in-use light duty vehicles, made up of the most representative emission control technologies in Mexico City: no catalyst, oxidative catalyst, and three way catalyst. Exhaust regulated (CO, NOx, and hydrocarbons) and toxic (benzene, formaldehyde, acetaldehyde, and 1,3 butadiene) emissions were evaluated for MTBE (5 vol %)- and ethanol (3, 6, and 10 vol %)-gasoline blends. The most significant overall emissions variations derived from the use of 6 vol % ethanol (relative to a 5% MTBE base gasoline) were 16% decrease in CO, 28% reduction in formaldehyde, and 80% increase in acetaldehyde emissions. A 26% reduction in CO emissions from the oldest fleet (< MY 1991, without catalytic converter), which represents about 44% of the in-use light duty vehicles in Mexico city, can be attained when using 6 vol% ethanol-gasoline, without significant variation in hydrocarbons and NOx emissions, when compared with a 5% vol MTBE-gasoline. On the basis of the emissions results, an estimation of the change in the motor vehicle emissions of the metropolitan area of Mexico city was calculated for the year 2010 if ethanol were to be used instead of MTBE, and the outcome was a considerable decrease in all regulated and toxic emissions, despite the growing motor vehicle population. PMID- 11393967 TI - Thiols in coastal waters of the western North Sea and English Channel. AB - Thiols were determined in coastal waters of the western North Sea and English Channel. Detection was carried out on-board ship on-line by flow-analysis with detection by cathodic stripping voltammetry and calibration with thiourea. The thiol concentrations ranged from 0.70 to 3.60 nM (thiourea equivalents) and were found to vary over a relatively short distance perpendicular to the coast. Low concentrations in the area of greatest estuarine input (the Humber-Wash area) indicated that the thiols did not originate from low-salinity waters. Instead, variations in the thiol concentration were found to parallel those of chlorophyll. This correlation was confirmed by depth profiles which showed a trend for the thiols similar to that of chlorophyll. The data demonstrates that thiols are more widespread than anticipated, and that marine phytoplankton is an important source of the thiols. In view of their known ability to bind with metals, these data indicate that the thiols could be an important candidate to act as such ligands in the marine system. The measurements gave no evidence for the presence of sulfide in these waters which means that it must be present at less than 20% of the detected thiol levels. PMID- 11393968 TI - Highly polar organic compounds present in wood smoke and in the ambient atmosphere. AB - Fine particulate matter emitted during wood combustion is known to contribute a significant fraction of the total fine aerosol concentration in the atmosphere of both urban and rural areas. In the present study, additional organic compounds that may act as wood smoke tracers in the atmosphere are sought. Polar organic compounds in wood smoke fine particulate matter are converted to their trimethylsilyl derivatives and analyzed by gas chromatography/mass spectrometry. Silylation enables the detection of n-alkanols, plant sterols, and a number of compounds derived from wood lignin that have not previously been reported in wood smoke samples, as well as levoglucosan and related sugar anhydrides formed during the combustion of cellulose. The concentrations of these compounds measured in source emissions are compared to the concentrations in atmospheric fine particle samples collected at a rural background site and at two urban sites in California's San Joaquin Valley. On the basis of this analysis, the sugar anhydrides galactosan and mannosan can be listed along with levoglucosan as being among the most abundant organic compounds detected in all samples. PMID- 11393969 TI - Hexachlorocyclohexane (HCH) isomers and chiral signatures of alpha-HCH in the Arctic marine food web of the Northwater Polynya. AB - Concentrations of hexachlorocyclohexane (HCH) isomers (alpha, beta, and gamma) and enantiomer fractions (EFs) of alpha-HCH were determined in the Northwater Polynya Arctic marine food web. Relative food web structure was established using trophic level models based on organic delta 15N values. Concentrations of HCH in the samples collected, including water, sediment, benthic invertebrates (four species), pelagic zooplankton (six species), Arctic cod, seabirds (seven species), and ringed seal, were in the range previously reported for the Canadian Arctic. The relative proportion of the HCH isomers varied across the food web and appeared to be related to the biotransformation capacity of each species. For invertebrates and fish the biomagnification factors (BMFs) of the three isomers were > 1 and the proportion of each isomer and the EFs of alpha-HCH were similar to water, suggesting minimal biotransformation. Seabirds appear to readily metabolize gamma- and alpha-HCH based on low BMFs for these isomers, high proportions of beta-HCH (62-96%), and high EFs (0.65-0.97) for alpha-HCH. The alpha- and beta-HCH isomers appear to be recalcitrant in ringed seals based on BMFs > 1 and near racemic EFs for alpha-HCH. The beta isomer appears to be recalcitrant in all species examined and had an overall food web magnification factor of 3.9. EFs of alpha-HCH and the proportion of beta-HCH in sigma-HCH in the food web were highly correlated (r2 = 0.92) suggesting that EFs were a good indicator of a species capability to biotransform alpha-HCH. PMID- 11393970 TI - New structural information on a humic acid from two-dimensional 1H-13C correlation solid-state nuclear magnetic resonance. AB - New information on the chemical structure of a peat humic acid has been obtained using a series of two-dimensional 1H-13C heteronuclear correlation solid-state NMR (HETCOR) experiments with different contact times and with spectral editing by dipolar dephasing and 13C transverse relaxation filtering. Carbon-bonded methyl groups (C-CH3) are found to be near both aliphatic and O-alkyl but not aromatic groups. The spectra prove that most OCH3 groups are connected directly with the aromatic rings, as is typical in lignin. As a result, about one-third of the aromatic C-O groups is not phenolic C-OH but C-OCH3. Both protonated and unprotonated anomeric O-C-O carbons are identified in the one- and two dimensional spectra. COO groups are found predominantly in OCHn-COO environments, but some are also bonded to aromatic rings and aliphatic groups. All models of humic acids in the literature lack at least some of the features observed here. Compositional heterogeneity was studied by introducing 1H spin diffusion into the HETCOR experiment. Comparison with data for a synthetic polymer, polycarbonate, indicates that the separation between O-alkyl and aromatic groups in the humic acid is less than 1.5 nm. However, transverse 13C relaxation filtering under 1H decoupling reveals heterogeneity on a nanometer scale, with the slow-relaxing component being rich in lignin-like aromatic-C-O-CH3 moieties and poor in COO groups. PMID- 11393971 TI - Time-resolved characterization of diesel particulate emissions. 2. Instruments for elemental and organic carbon measurements. AB - The measurement of elemental carbon (EC) and organic carbon (OC) mass for particles emitted by diesel vehicles is currently accomplished using particle collection on filters, followed by analysis using the thermal/optical reflectance carbon analysis method (TOR) or one of its variations. Such filter methods limit time resolution to a minimum of several minutes, making it impossible to study emissions during transient operating conditions. Testing of five different measurement methods has demonstrated that fast response measurement of diesel exhaust particulate EC and OC concentrations, consistent with TOR filter measurements, is feasible using existing technology. EC mass concentrations are best measured through determination of particulate light absorption with a photoacoustic instrument or determination of light extinction with a smoke meter. The photoacoustic instrument has the better dynamic range and sensitivity, whereas the smoke meter is a simpler instrument. Fast response OC measurements cannot be made with any single instrument tested. However, a combination of real time weighing as implemented in the tapered element oscillating microbalance with the photoacoustic instrument has been shown to be capable of determining OC concentrations with good time response. The addition of a nephelometer to the OC measurement could potentially improve time resolution, freedom from interferences, and sensitivity. PMID- 11393972 TI - Polycyclic aromatic hydrocarbons identified in soot extracts from domestic coal burning stoves of Henan Province, China. AB - Using high-pressure liquid chromatography with ultraviolet-visible diode-array detection, we have analyzed polycyclic aromatic hydrocarbons (PAH) in the dichloromethane extracts of soot deposits from coal-burning stoves in several homes of Henan Province, China--including Linxian County, where esophageal cancer rates are some of the highest in the world. Thirty-two individual polycyclic aromatic compounds, ranging in size from three to eight fused aromatic rings, have been unequivocally identified among the soot extract components--including 20 benzenoid PAH, 6 fluoranthene benzologues, 1 cyclopentafused PAH, 1 indene benzologue, 3 oxygenated PAH, and 1 ring-sulfur-containing aromatic. Most of the identified compounds have been observed before among the products of laboratory coal pyrolysis experiments, but two of the components, the six-ring C24H14 napthol[1,2-b]fluoranthene and the eight-ring C30H16 tribenzo[e,ghi,k]perylene, have never before been documented as products of coal in any system. All of the Henan coal soot extracts are remarkably similar qualitatively in that they contain the same set of identified PAH, but absolute levels of individual species vary by up to 5 orders of magnitude, from sample to sample. The bulk of the identified component mass in all of these soot extracts lies in the five- and six ring PAH--the largest single class being the family of five-ring C20H12 isomers, to which the samples' most abundant components, benzo[b]fluoranthene and benzo[e]pyrene, belong. The five- and six-ring PAH also account for the majority of the samples' known mutagens. The three strong mutagens identified in these soot samples are the C20H12 benzo[a]-pyrene and two C24H14 PAH, dibenzo[a,e]pyrene and naphtho-[2,1-a]pyrene. Seven moderate mutagens are found among the C20H12, C22H12, C22H14, and C24H14 PAH. A major class of mutagens, the cyclopenta-fused PAH, appears to be absent from these samples, but our detection of an oxidation product of the major mutagen cyclopenta[cd]- pyrene--itself mutagenic--suggests that these soot deposits may contain additional mutagenic cyclopentafused PAH oxidation products as well. PMID- 11393973 TI - Determination of silver speciation in natural waters. 1. Laboratory tests of Chelex-100 chelating resin as a competing ligand. AB - Batch and column experiments were performed to investigate the suitability and chemical characteristics of Chelex-100 for use as a competing ligand in ionic silver (Ag(I)) speciation determinations in natural waters. A conditional stability constant (Kcond) for Ag+ chelation by iminodiacetate groups on the surface of Chelex resin was determined by fitting results of batch and column experiments with an equilibrium speciation model. Results of experiments in which Chelex competed with cyanide ion and thiosulfate ion for aqueous Ag+ were fitted well by a model in which log Kcond(Ag-Chelex) was set to 7.2. This value is similar to literature equilibrium constants for a 1:1 Ag(+)-EDTA chelate. In batch experiments with Chelex, equilibration times of 24 h were found to be sufficient to bring samples close to equilibrium. Effects of resin counterion and total Ag(I) concentration on extent of Ag(I) chelation were found to be minor. Effect of pH on Ag(I) chelation was minor over a range of 6-10. Column experiments (detention time = 6 s, empty-column basis) in which thiosulfate competed with Chelex for Ag(I) gave similar results to batch experiments with thiosulfate. This implies that batch and column experiments could be compared to explore ligands in natural water systems with different rates of dissociation. PMID- 11393974 TI - Determination of silver speciation in natural waters. 2. Binding strength of silver ligands in surface freshwaters. AB - Three competing ligand methods were compared to determine characteristics of Ag(I) complexation by dissolved and colloidal ligands present in three rivers and one sewage treatment plant effluent. Iminodiacetate groups on Chelex-100 resin (used in batch and column experiments) and diethyldithiocarbamate (DDC) were used as competing ligands. Results of batch Chelex and DDC competition experiments show good agreement with regard to relative extent of Ag binding by natural ligands among the three river systems. Results of both methods also show a possible correlation between extent of Ag(I) complexation and organic matter concentration and/or Fe concentration. Fraction of Ag(I) associated with Chelex in both batch and column Chelex experiments was similar in each of the four systems tested, indicating that lability of Ag complexes does not change significantly on time scales ranging from seconds to 24 h. Results of Chelex and DDC competition were compared using a model based on a hypothetical single natural ligand. Under the experimental conditions used, this model quantified Ag(I) complexes with log Kcond values from approximately 12 to 14. For the three rivers studied, ligands with silver-association characteristics similar to those of reduced sulfur groups (log K = 14-16) present at subnanomolar concentrations likely dominate Ag(I) speciation in these systems. A weaker ligand (e.g., log Kcond < 12) at concentrations > 0.7 nM dominated Ag(I) speciation in the treatment plant effluent. This may result from elevated concentration of metals that compete for reduced sulfur groups rather than from a lower total concentration of these groups. PMID- 11393975 TI - Identifying sources and mass balance of dioxin pollution in Lake Shinji Basin, Japan. AB - On the basis of congener-specific analysis of dioxins in a dated sediment core, the sources and behavior of dioxins in Lake Shinji Basin, Japan, were estimated. The dioxins in the core showed that their deposition in the lake increased rapidly during the 1960s, peaked in the early 1970s, and then decreased gradually. Principal component analysis of the congener-specific data showed that three major sources existed: pentachlorophenol (PCP), chloronitrophen (CNP), and combustion. PCP and CNP are paddy field herbicides used extensively in the basin. The time trends of source contributions were estimated by multiple regression analysis using the source profiles. The results revealed that dioxin emission from PCP and CNP herbicides was high in the 1960s and the early 1970s, respectively. The contributions from PCP, CNP, and combustion in recent surface sediment were about 68, 16, and 16% in terms of total amount of dioxins. From the decreasing trend of dioxin deposition in the lake after extensive herbicide use, the amount of dioxins that accumulated in the agricultural soil in the basin was estimated to have decreased by about 2%/yr or a half-life of about 35 yr, indicating that dioxin runoff from agricultural fields would continue for a long time. PMID- 11393976 TI - Further evidence for the existence of PCDD/Fs in the environment prior to 1900. AB - PCDD/Fs and PCBs have been analyzed in a series of archived soil samples collected from various depths during the 1800s and early 1900s. PCBs were not found in soil samples collected before 1900, whereas PCDD/Fs were present in concentrations between 43 and 110 pg/g in surface soils, and between 9 and 150 pg/g in soils collected from below the surface. The PCDD/F homologue patterns of all surface soils were consistent with each other. The homologue pattern of deeper soils altered with depth to one that was dominated by highly chlorinated PCDDs. The highest sigma(4-8)PCDD/F concentration (150 pg/g) was found in the deepest soil analyzed (230-250 cm below the surface). The cork from one of the storage bottles contained considerable quantities of both PCBs and PCDD/Fs. However, contamination of the soils, either by diffusion through the cork or by cork particles, was discounted on the basis that no PCBs were evident in the soil, and that the PCDD/F homologue pattern in the cork was very different to that found in the soil. Similar arguments were used to discount contamination of the soil by dust. A sample of ashed vegetation from the archive, that had no cork stopper, contained high concentrations of PCBs (78 ng/g), concentrations of mono- to tri-CDFs that were higher than in any of the soils (190 pg/g), but very low concentrations of sigma(4-8)PCDD/F (12 pg/g). This pattern of analytes was considered to be representative of contamination from store room air and was completely different from the pattern observed in the soils. Taken together these observations indicate that contamination during storage, or subsequent handling, is unlikely to have occurred in archived soil samples that were stored with cork and wax seal intact. The results imply surface soil sigma(4-8)PCDD/F concentrations of around 60 pg/g at Rothamsted (southeast England) in the late 1800s, compared with approximately 300 pg/g reported for rural UK soils in the 1990s. PMID- 11393977 TI - Design of an inventory system for the volatile organic compounds emitted by various activities. AB - In this study an inventory system was designed which fulfills the reporting requirements of the current European Union legislation for air emissions. Given the reporting requirements deriving from the EU Directives, International Conventions, and Inventory Methodologies developed at the EU level, there are significant differences in the required data in terms of industrial categories, definitions of point and area sources, whether TVOC or individual species must be determined, and expression of VOCs emissions. On the basis of this concept, an inventory system was structured that aims to comply with the framework set by the European Union for the evaluation of VOCs emissions from industrial activities and other facilities in Greece. This inventory can be very helpful to all the EU members states, as well as the Eastern European countries that are in pre accession phase, who must implement the European legislation concerning VOCs emissions. PMID- 11393978 TI - Organochlorine pesticide residues in archived UK soil. AB - Archived background soils ("Broadbalk', 1944-1986) and sludge-amended soils ("Luddington", 1968-1990), collected from long-term agricultural experiments in the UK, were analyzed for a range of organochlorine (OC) pesticides to establish trends over time. Concentrations typically ranged from 0.1 to 10 ng/g of soil (dry weight), with gamma-hexachlorocyclohexane (gamma-HCH), dieldrin, and p,p' DDE consistently having the highest concentrations. The trends in the Broadbalk background soils are largely consistent with usage patterns, with peak concentrations occurring in the 1960s for DDTs and between the 1960s and the 1980s for the other OCs. In the Luddington control and sludge-amended soils, several of the OCs show a significant decline in concentrations from the late 1960s to 1990, with half-lives ranging from approximately 7 years (alpha-HCH) to approximately 25 years (dieldrin). The sludge-amended plot received 125 tonnes of sludge per ha in 1968, which was mixed in to a depth of 15 cm. It appears that the sludge treatment had little effect on concentrations in the soil, with no significant difference between control soil and sludge-amended soil for most compounds, except for HCB, p,p'-DDE, and dieldrin. Enantiomeric fractions (EFs) of some chiral pesticides (alpha-HCH, cis- and trans-chlordane, and o,p'-DDT) were determined in the Luddington soils. Results reveal that enantioselective degradation of OC pesticides is occurring in these soils for trans-chlordane (TC) and cis-chlordane (CC). However, the depletion over time is not statistically significant, and there is no statistically significant difference between EFs in the control soil and sludge-amended soil. This indicates that enantioselective microbial degradation was not consistent over time and that the addition of sludge to soil did not significantly alter the enantiomeric preference of the microbial community. PMID- 11393979 TI - Platinum and rhodium concentrations in airborne particulate matter in Germany from 1988 to 1998. AB - Increases in platinum group element (PGE) concentrations in ambient air and dust since the introduction of automotive catalytic converters in 1988 is a cause of concern. Until now, data derived from engine-test bench experiments have provided the basis for the assessment of human health risks associated with PGE exposure. Such experiments have provided valuable information regarding emission data that has been used to estimate ambient exposure concentrations. However, these data are not necessarily representative of typical environmental PGE exposure levels and conditions. Data on measured environmental concentrations is needed to provide a more adequate basis for the assessment of exposure and related risks. Twenty air and airborne-dust samples were provided by the Umweltbundesamt (Federal Environmental Agency, Germany) in the years 1988, 1989, 1992, 1997, and 1998. The samples were collected in Frankfurt/Main and the adjacent city of Offenbach. For this, 11 to 80 m3 of air were filtered over a 24-72 h period using a vacuum. Glass-fiber filters were used to collect samples. Sample platinum and rhodium concentrations were determined using adsorptive voltammetry. Although the number of samples collected in different years is limited, the results indicate a trend toward continuous increases in ambient concentrations of these metals between 1988 and 1998. Specifically, there were 46- and 27-fold increases in Pt and Rh concentrations, respectively. Despite these observed increases, the Pt concentrations measured (i.e., 147 pg/m3 on average, with a maximum of 246 pg/m3 in 1998) fell far below 15,000 pg/m3, which has been suggested as a guidance value (i.e., exposure at this level would be expected to be without appreciable health risk). The results of a particle-size distribution analysis of one sample (8-step impactor) that was collected 150 m away from a street show that approximately 75% of Pt and 95% of Rh occurs in association with large particulate matter of > 2 microns, with concentrations reaching a maximum in particles of 4.7 to 5.8 microns. The remaining 25% of Pt and 5% of Rh is present in fine particulate matter of < 2 microns. An approximate 10% of Pt and < 38% of Rh in airborne particles was found to be soluble in 0.1 molar HCl. Further, the results indicate that most of the emitted PGE particles from automotive catalytic converters, particularly those bound to fine particulate matter, are capable of being airborne. As a result, PGEs are not only present in areas close to emissions (e.g., roads), but can be transported over longer distances. PMID- 11393980 TI - Release of hydrogen chloride from combustibles in municipal solid waste. AB - Study of the inorganic chlorides in municipal solid waste (MSW) shows that the main source of inorganic chlorides in MSW is food. The main organic source of HCl emission from MSW is plastic. But wood, textiles, and food also produce a large amount of HCl when they are combusted. Each combustible shows a different HCl releasing temperature range. At 973 K, there are 30-70% of the total chlorine left in the char of each combustibles in MSW. PMID- 11393981 TI - Adsorption of hydroxy pyridines and quinolines at the metal oxide-water interface: role of tautomeric equilibrium. AB - We have identified compound physical-chemical properties and structural criteria required to facilitate the adsorption of hydroxypyridines, quinolines, and pyrimidines onto aluminum(III), iron(III), and titanium(IV) oxides. We achieved this by measuring the adsorption behavior of a number of structurally related compounds, each with a nearly isolated variation in molecular structure, and systematically evaluating the effect of structural change on the nature and extent of adsorption. We find that the adsorption at the metal oxide-water interface is significant only when the oxo-hydroxy (keto-enol) tautomeric equilibrium in aqueous solution favors the hydroxy tautomer. Lack of adsorption of the oxo tautomer can be attributed to the absence of favorable electrostatic interactions between the compound and the surface, the absence of ligand groups capable of surface complexation, and the presence of strong intermolecular hydrogen bonding between ligand groups (carbonyl and amide) and water molecules. We find that adsorption of hydroxy tautomers with ortho-substituted cyclic N and OH groups (such as trichloropyridinol) likely occurs via nonspecific electrostatic interactions, while non-ortho-substituted hydroxy tautomers (such as 8-hydroxyquinoline) and compounds such as (2-hydroxy-methylpyridine) likely adsorb via surface complexation. This study demonstrates that for compounds classified as tautomeric heterocycles it is imperative to accurately represent the predominant tautomer in aqueous solution so that the appropriate physical chemical properties and reactivities are utilized in the evaluation of environmental fate. PMID- 11393982 TI - Biologically-enhanced removal of PCE from NAPL source zones. AB - The influence of dechlorinating microorganisms on PCE and its reduction products in a residual nonaqueous phase liquid (NAPL) source zone was investigated. Experiments were conducted in upflow columns containing glass beads (diameters 500-750 microns) contaminated with a residual NAPL consisting of tridecane and labeled 14C-PCE. Three columns were inoculated with a mixed PCE-dechlorinating culture, that was fed electron donor (pyruvate) at concentrations of 25, 100, and 250 mM. Pyruvate was fermented in all columns with essentially no methanogenic activity. Comparisons between actively dechlorinating columns and abiotic-PCE columns demonstrated that dechlorination resulted in an increase in total PCE removal, up to a factor of 16 over dissolution. PCE was sequentially reduced to trichloroethene, cis-dichloroethene, and vinyl chloride without ethene formation over the experimental period in the two columns operated at the lower electron donor levels. Total chlorinated ethenes removal for the columns that retained dechlorinating populations was enhanced from 5.0 to 6.5 times over the removal that would have resulted from dissolution alone. The system fed the highest pyruvate levels, interestingly, lost dechlorinating activity early in the experiment. PMID- 11393983 TI - Spectrophotometric assay of POE nonionic surfactants and its application to surfactant sorption isotherms. AB - The operational range and suitability toward environmental samples of an iodine iodide (I-I) assay for nonionic surfactants were assessed. The I-I assay provides a rapid and repeatable method for determining aqueous nonionic surfactant concentrations. Through a systematic examination of surfactant structure, the operational range of the assay was shown to be on the order of 10(-6) to 10(-3) MEO, where the concentration unit MEO is defined as the molar surfactant concentration multiplied by the number of ethylene oxide units in the surfactant molecule. For environmental samples, it was shown that the I-I assay can be applied to measurement of surfactant sorption isotherms to aquifer sands and bacteria cultures. A potential limitation of the I-I assay is interference with humic acids, with the magnitude of the interference dependent on the concentration of humic acids present. The main benefit of the I-I assay is that its high accuracy and ease of application allows measurement of low levels of surfactant sorption. Surfactant sorption to aquifer sand could be measured down to the range of 10(-9) mol/g. PMID- 11393984 TI - Electrochemical and spectroscopic study of arsenate removal from water using zero valent iron media. AB - This study investigated the mechanisms involved in removing arsenate from drinking water supplies using zero-valent iron media. Batch experiments utilizing iron wires suspended in anaerobic arsenate solutions were performed to determine arsenate removal rates as a function of the arsenate solution concentration. Corrosion rates of the iron wires were determined as a function of elapsed time using Tafel analysis. The removal kinetics in the batch reactors were best described by a dual-rate model in which arsenate removal was pseudo-first-order at low concentrations and approached zero-order in the limit of high arsenate concentrations. The presence of arsenate decreased iron corrosion rates as compared to those in blank 3 mM CaSO4 background electrolyte solutions. However, constant corrosion rates were attained after approximately 10 days elapsed, indicating that the passivation processes had reached steady state. The cathodic Tafel slopes were the same in the arsenate and the blank electrolyte solutions. This indicates that water was the primary oxidant for iron corrosion and that arsenate did not directly oxidize the iron wires. The anodic Tafel slopes were greater in the arsenate solutions, indicating that arsenate formed complexes with iron corrosion products released at anodic sites on the iron surfaces. Ion chromatography analyses indicated that there was no measurable reduction of As(V) to As(III). X-ray absorption spectroscopy analyses indicated that all arsenic associated with the zero-valent iron surfaces was in the oxidation state. Interatomic arsenic-iron distances determined from EXAFS analyses were consistent with bidentate corner-sharing among arsenate tetrahedra and iron octahedra. Results from this study show that under conditions applicable to drinking water treatment, arsenate removal by zero-valent iron media involves surface complexation only and does not involve reduction to metallic arsenic. PMID- 11393985 TI - Influence of the nonionic surfactant Brij 35 on the bioavailability of solid and sorbed dibenzofuran. AB - The effect of the nonionic surfactant Brij 35 on the bioavailability of solid and Teflon-sorbed dibenzofuran for Sphingomonas sp. strain HH19k was studied in simple model systems. Growth with dibenzofuran and dibenzofuran-specific oxygen uptake in surfactant-free media and with Brij 35 above the critical micelle concentration (cmc) were compared with dissolution and desorption in the absence of bacteria. Brij 35 accelerated dissolution and biodegradation of solid dibenzofuran by a factor of 2. It also enhanced the initial desorption rate of dibenzofuran from Teflon by this factor. Continuously decreasing desorption rates were attributed to slow diffusion of dibenzofuran inside Teflon, leading to depletion of dibenzofuran in the exterior of the Teflon particles. Surprisingly, Brij 35 slowed the initial biodegradation of desorbing dibenzofuran. We propose two processes that led to low bioavailability of sorbed dibenzofuran in the presence of surfactant. First, desorbing dibenzofuran rapidly accumulated in surfactant micelles, leading to reduced truly water-dissolved dibenzofuran concentration as the factor controlling the biodegradation rate. Second, Brij 35 suppressed the contact between bacteria and Teflon. This increased the average diffusion distance of dibenzofuran to the bacteria, which in turn flattened the gradient of the dissolved dibenzofuran concentration between the sorbent and the cells as the driving force for desorption. PMID- 11393986 TI - Fate of linear alkylbenzenes released to the coastal environment near Boston Harbor. AB - Linear alkylbenzenes (LABs) were used to assess the fates of hydrophobic organic compounds (HOCs) released to a large urban harbor and the adjoining offshore waters. We found that particulate concentrations of the individual C12 LAB isomers in 1996 summertime surface waters decreased from 1 pM in Boston Harbor to 20-200 fM in coastal Massachusetts and Cape Cod Bays. Levels fell to only a few fM in offshore Gulf of Maine locations. These observations were consistent with municipal wastewater in Boston Harbor as the predominant input followed by dispersal via known circulation patterns in this region. Phase-dependent removal rate coefficients for flushing, vertical scavenging, volatilization, photodegradation, and biodegradation of individual LAB isomers were constrained from literature, field observations, and laboratory experiments and combined with estimates of wastewater release rates into a predictive 3-box model. Vertical scavenging, biodegradation, and flushing were predicted to be the most important fate processes for C12 LABs in the Boston Harbor-MA Bay-Cape Cod Bay flow system with about 1% of the harbor releases "surviving" passage. For HOCs such as the relatively bio-recalcitrant LAB, 6-phenyldodecane, it appears that we are at present able to predict the coastal fate of harbor-introduced HOCs in this system within a factor of 2. Contrary to expectations from biodegradation experiments, the ratio of internal-to-external (I/E) LAB isomers decreased offshore in both water and sediment samples, suggesting we are "missing" an important process affecting LAB fates. PMID- 11393987 TI - Generic NICA-Donnan model parameters for proton binding by humic substances. AB - Forty-nine datasets consisting of literature and experimental data for proton binding by fulvic and humic acids have been analyzed using the NICA-Donnan model. The model successfully described the behavior of the individual datasets with a high degree of accuracy and highlighted the differences in site density and binding affinity between fulvic acids (FA) and humic acids (HA) while demonstrating their strong similarities. The data have also been used to derive generic model descriptions of proton binding by FA and HA that can be used for modeling in the absence of specific parameter sets for the particular humic substance of interest. These generic parameters can provide estimates of the amount of proton binding by a wide variety of humic substances to within approximately +/- 20% under any given conditions. The maximum site density for protons was 7.74 and 5.70 equiv kg-1 for a generic FA and HA, respectively. The recommended generic NICA-Donnan parameter values for FA are b = 0.57, Qmax1,H = 5.88, log KH1 = 2.34, mH1 = 0.38, Qmax2,H = 1.86, log KH2 = 8.6, and mH2 = 0.53; for HA the values are b = 0.49, Qmax1,H = 3.15, log KH1 = 2.93, mH1 = 0.50, Qmax2,H = 2.55, log KH2 = 8.0, and mH2 = 0.26. PMID- 11393988 TI - Source apportionment of fine particulate matter by clustering single-particle data: tests of receptor model accuracy. AB - The source apportionment accuracy of a neural network algorithm (ART-2a) is tested on the basis of its application to synthetic single-particle data generated by a source-oriented aerosol processes trajectory model that simulates particle emission, transport, and chemical reactions in the atmosphere. ART-2a successfully groups particles from the majority of sources actually present, when given complete data on ambient particle composition at monitoring sites located near the emission sources. As particles age in the atmosphere, accumulation of gas-to-particle conversion products can act to disguise the source of the primary core of the particles. When ART-2a is applied to synthetic single-particle data that are modified to simulate the biases in aerosol time-of-flight mass spectrometry (ATOFMS) measurements, best results are obtained using the ATOFMS dual ion operating mode that simultaneously yields both positive and negative ion mass spectra. The results of this study suggest that the use of continuous single particle measurements coupled with neural network algorithms can significantly improve the time resolution of particulate matter source apportionment. PMID- 11393989 TI - Modeling of fugitive dust emission for construction sand and gravel processing plant. AB - Due to rapid economic development in Taiwan, a large quantity of construction sand and gravel is needed to support domestic civil construction projects. However, a construction sand and gravel processing plant is often a major source of air pollution, due to its associated fugitive dust emission. To predict the amount of fugitive dust emitted from this kind of processing plant, a semiempirical model was developed in this study. This model was developed on the basis of the actual dust emission data (i.e., total suspended particulate, TSP) and four on-site operating parameters (i.e., wind speed (u), soil moisture (M), soil silt content (s), and number (N) of trucks) measured at a construction sand and gravel processing plant. On the basis of the on-site measured data and an SAS nonlinear regression program, the expression of this model is E = 0.011.u2.653.M 1.875.s0.060.N0.896, where E is the amount (kg/ton) of dust emitted during the production of each ton of gravel and sand. This model can serve as a facile tool for predicting the fugitive dust emission from a construction sand and gravel processing plant. PMID- 11393990 TI - Comparison of two floor mat lead dust collection methods and their application in pre-1950 and new urban houses. AB - This study investigated commercial floor mats as an alternative method to assess lead in residential dust in inner-city houses. Mats were placed for 3 weeks in interior entry-ways of 34 row houses built before 1950 and 17 new row houses in Baltimore City. A high volume sampler (an HVS3 floor model cyclone-based vacuum) and a hand-held portable cyclone sampler were used in the laboratory to collect side-by-side samples of mat dust. Both devices yielded comparable estimates of lead dust deposition, dust lead concentration, and dust deposition on field mat samples and had similar sampling efficiencies on mats spiked with various types of standard reference materials. The older houses had significantly higher daily lead dust deposition (mean = 130 micrograms/ft2/day by HVS3) than the newer houses (mean = 9 micrograms/ft2/day by HVS3), due to higher dust lead concentrations (mean = 1149 ppm vs mean = 107 ppm by HVS3) and not to differences in daily dust deposition (mean = 118 mg/ft2/day vs mean = 87 mg/ft2/day by HVS3) [corrected]. Mats were found to be a feasible method for the collection of dust that has accumulated for a known amount of time. Current wipe and vacuum methods do not allow for the estimation of dust deposition rates. Further research is needed to understand the role of floor mats as a risk assessment tool. PMID- 11393991 TI - Selective trace level analysis of phenolic compounds in water by flow injection analysis--membrane introduction mass spectrometry. AB - Flow injection analysis coupled with membrane introduction mass spectrometry (FIA MIMS) with on-line derivatization is shown to allow fast, accurate, nearly interference-free, and sensitive (low microgram/L) quantitation of phenolic compounds in water. On-line FIA derivatization of the phenolic compounds is performed by acetic anhydride acetylation in a K2CO3-buffered alkaline medium. The phenol acetates so formed efficiently permeate a silicone membrane and are directly transferred to the mass spectrometer, in which they are analyzed with selectivity and high sensitivity via selected ion monitoring. FIA-MIMS analysis was performed for aqueous solutions of phenol, 2-methylphenol, 4-chlorophenol, 4 chloro-3-methylphenol, 2,4-dichlorophenol, and 2,4,6-trichlorophenol, and detection limits in the 0.5-20 micrograms/L (ppb) range were observed for an analytical frequency of six samples/h. FIA-MIMS for phenolic compound analysis is considerably less time-consuming and labor intensive than most chromatographic methods based on liquid-liquid extraction and preconcentration procedures and is therefore applicable for on-line and in-situ monitoring of phenols in wastewaters and in the environment. FIA-MIMS employing acetic anhydride derivatization is also virtually free of interferences since it combines chemical, membrane, and enhanced MS selectivity; hence quantitation of phenolic compounds can be performed in the presence of congeners. PMID- 11393992 TI - Using time- and size-resolved particulate data to quantify indoor penetration and deposition behavior. AB - Because people spend approximately 85-90% of their time indoors, it is widely recognized that a significant portion of total personal exposures to ambient particles occurs in indoor environments. Although penetration efficiencies and deposition rates regulate indoor exposures to ambient particles, few data exist on the levels or variability of these infiltration parameters, in particular for time- and size-resolved data. To investigate ambient particle infiltration, a comprehensive particle characterization study was conducted in nine nonsmoking homes in the metropolitan Boston area. Continuous indoor and outdoor PM2.5 and size distribution measurements were made in each of the study homes over weeklong periods. Data for nighttime, nonsource periods were used to quantify infiltration factors for PM2.5 as well as for 17 discrete particle size intervals between 0.02 and 10 microns. Infiltration factors for PM2.5 exhibited large intra- and interhome variability, which was attributed to seasonal effects and home dynamics. As expected, minimum infiltration factors were observed for ultrafine and coarse particles. A physical-statistical model was used to estimate size specific penetration efficiencies and deposition rates for these study homes. Our data show that the penetration efficiency depends on particle size as well as home characteristics. These results provide new insight on the protective role of the building shell in reducing indoor exposures to ambient particles, especially for tighter (e.g., winterized) homes and for particles with diameters greater than 1 micron. PMID- 11393993 TI - Emission factors of carbon monoxide and size-resolved aerosols from biofuel combustion. AB - This study reports emission factors of carbon monoxide and size-resolved aerosols from combustion of wood, dung cake, and biofuel briquette in traditional and improved stoves in India. Wood was the cleanest burning fuel, with higher emissions of CO from dung cake and particulate matter from both dung cake and briquette fuels. Combustion of dung cake, especially in an improved metal stove, resulted in extremely high pollutant emissions. Instead, biogas from anaerobic dung digestion should be promoted as a cooking fuel for public health protection. Pollutant emissions increased with increasing stove thermal efficiency, implying that thermal efficiency enhancement in the improved stoves was mainly from design features leading to increased heat transfer but not combustion efficiency. Compared to the traditional stove, the improved stoves resulted in the lower pollutant emissions on a kW h-1 basis from wood combustion but in similar emissions from briquette and dung cake. Stove designs are needed with good emissions performance across multiple fuels. Unimodal aerosol size distributions were measured from biofuel combustion with mass median aerodynamic diameters of 0.5-0.8 micron, about a factor of 10 larger than those from fossil fuel combustion (e.g. diesel), with potential implications for lung deposition and health risk. PMID- 11393994 TI - Phytochelatins are bioindicators of atmospheric metal exposure via direct foliar uptake in trees near Sudbury, Ontario, Canada. AB - Plants produce phytochelatins in response to copper and nickel, the primary metal pollutants emitted by the dominant smelting operation in Sudbury. Copper and nickel concentrations in soils decline sharply with distance from this facility, primarily as a result of early smelting practices. Phytochelatin concentrations in Sudbury-area trees, however, do not correlate with metal levels in soils. Rather, phytochelatin production in tree leaves is driven by metals currently released to the atmosphere through the 381 m emissions stack. Phytochelatin concentrations in the foliage of three tree species growing in situ are highest 20-30 km from the stack, correlated with maximum acid-leachable concentrations of deposited copper and nickel. Similar results observed in potted trees placed adjacent to indigenous trees confirm that aerially deposited metals are the source of current metal stress patterns. The addition of peat moss "filters" to potted soils did not alter this response, indicating that direct foliar metal uptake is responsible. PMID- 11393995 TI - Solar oxidation and removal of arsenic at circumneutral pH in iron containing waters. AB - An estimated 30-50 million people in Bangladesh consume groundwater with arsenic contents far above accepted limits. A better understanding of arsenic redox kinetics and simple water treatment procedures are urgently needed. We have studied thermal and photochemical As(III) oxidation in the laboratory, on a time scale of hours, in water containing 500 micrograms/L As(III), 0.06-5 mg/L Fe(II,III), and 4-6 mM bicarbonate at pH 6.5-8.0. As(V) was measured colorimetrically, and As(III) and As(tot) were measured by As(III)/As(tot) specific hydride-generation AAS. Dissolved oxygen and micromolar hydrogen peroxide did not oxidize As(III) on a time scale of hours. As(III) was partly oxidized in the dark by addition of Fe(II) to aerated water, presumably by reactive intermediates formed in the reduction of oxygen by Fe(II). In solutions containing 0.06-5 mg/L Fe(II,III), over 90% of As(III) could be oxidized photochemically within 2-3 h by illumination with 90 W/m2 UV-A light. Citrate, by forming Fe(III) citrate complexes that are photolyzed with high quantum yields, strongly accelerated As(III) oxidation. The photoproduct of citrate (3 oxoglutaric acid) induced rapid flocculation and precipitation of Fe(III). In laboratory tests, 80-90% of total arsenic was removed after addition of 50 microM citrate or 100-200 microL (4-8 drops) of lemon juice/L, illumination for 2-3 h, and precipitation. The same procedure was able to remove 45-78% of total arsenic in first field trials in Bangladesh. PMID- 11393996 TI - Radiolytic and thermal dechlorination of organic chlorides adsorbed on molecular sieve 13X. AB - Reductive dechlorination of chlorobenzene (PhCl), trichloroethylene (TCE), tetrachloroethylene (PCE), 1- and 2-chlorobutanes, chloroform, carbon tetrachloride, and 1,1,1- and 1,1,2-trichloroethanes adsorbed on molecular sieve 13X was investigated. The molecular sieve adsorbing the organic chlorides was irradiated with gamma-rays, heated, or allowed to stand at room temperature in a sealed ampule and was then soaked in water. The dechlorination yields were determined from the Cl- concentrations of the supernatant aqueous solutions. It was found that the chlorinated alkanes adsorbed on the molecular sieve are readily dechlorinated on standing at room temperature. The dechlorination at room temperature was limited for TCE and PCE. PhCl was quite stable even at 200 degrees C. gamma-Radiolysis was examined for PhCl, TCE, and PCE at room temperature. The radiation chemical yields of the dechlorination, G(Cl-), were 1.9, 40, and 30 for PhCl, TCE, and PCE, respectively. After 5 h of heating at 200 degrees C, the dechlorination yields for TCE and PCE were 24.5 and 4.3%, respectively. TCE is much more reactive than PCE in the thermal dechlorination, whereas their radiolytic dechlorination yields are comparable. The pH of the supernatant solutions decreased along with the dechlorination. PMID- 11393997 TI - [Strategy of treatment for liver metastasis from colorectal cancer]. PMID- 11393998 TI - [Genetic changes in liver metastasis of colorectal cancer and their clinical application]. AB - It has been confirmed that several genes are involved in each step of liver metastasis of colorectal cancer. This article reviews recent highlights in this field. The expression of c-erbB2 or Rho protein in colorectal cancer tissues correlates closely with liver metastasis. Suppressor genes such as nm23, DCC, and DPC4 may play a role in the suppression of liver metastasis. On the other hand, the E-cadherin-catenin system, carbohydrate chains, selectin, and variant CD44 are known to play an important role in cells migration from the primary lesion, the adhesion of tumor cells to endothelial cells, and cell motility. These adhesion molecules may be a biological marker of liver metastasis. In addition, treatment targeting these genes will be a potent therapy for liver metastasis in the future. PMID- 11393999 TI - [Extracellular matrix degradation enzymes: important factors in liver metastasis of colorectal cancer and good targets for anticancer metastatic therapy]. AB - Extracellular matrix (ECM) degradation enzymes, such as matrix metalloproteinases (MMPs) and plasminogen activators, are important factors in cancer invasion and metastasis, because invasion and metastasis of cancer cells require destruction of mesenchymal collagen or the endothelial basement membrane. Moreover, recent studies have shown that ECM degradation enzymes play important roles in cancer cell proliferation, cancer escape from the immune system, and tumor angiogenesis. ECM degradation enzymes, especially some MMPs, are good targets for anticancer metastatic therapy. Numerous anti-MMP agents have been developed and phase III clinical trials in advanced cancers ongoing. Successful control of MMPs induced by cancer cells will prevent liver metastasis of colorectal cancer. PMID- 11394000 TI - [Significance of angiogenesis and clinical application of anti-angiogenesis]. AB - Angiogenesis is essential for tumor growth and metastasis and depends upon the production of angiogenic factors by host and/or tumor cells. Increased vascularity may allow not only an increase in tumor growth but also a greater chance for hematogenous metastasis. We have already reported that vessel density and vascular endothelial growth factor (VEGF) expression are higher in metastatic tumors than in nonmetastatic tumors and that VEGF and its receptor, the KDR ligand/receptor system, also correlate with metastasis. Therefore the anti-VEGF antibody and VEGF receptor antagonist are potential targets for antiangiogenesis therapy in colon cancer. Clinical trials of such agents are continuing to phase II/III in the USA and Europe. In this paper, we introduce data on antiangiogenesis agents in the treatment of metastatic colorectal cancer and point out that the strategy for antiangiogenesis is not tumor shrinkage but tumor dormancy. PMID- 11394001 TI - [Surgical treatment of liver metastases from colorectal cancer patient selection and oncological outcome]. AB - The indications for surgical resection of liver metastases from colorectal cancer remain controversial. Clinical, pathological, and outcome data for 418 consecutive patients undergoing hepatectomy between 1984 and 1998 were examined. The over-all 5-year survival rate was 42%, and the 10-year survival rate was 31%. The 5-year survival rate of patients with four or more nodules was 24%, with 20 patients surviving for more than 5 years. Five factors were found to be significant and independent predictors of poor long-term outcome by multivariate analysis. The preoperative scoring system reported by Fong et al was double checked in our 418 patients and was found to be useful to predict long-term outcome after hepatectomy. It is clear that liver resection alone has limitations. Therefore clinical risk scoring (CRS) and effective intravenous systemic chemotherapy to prevent recurrence both in the remaining liver and lung should be established to improve survival outcome in patients with poor prognostic factors after liver resection. PMID- 11394002 TI - [Efficacy of local ablation therapy for liver metastasis from colorectal cancer- radiofrequency ablation and microwave coagulation therapy]. AB - We evaluated the efficacy of local ablation therapy in 40 patients with liver metastases from colorectal cancer. Radiofrequency ablation (RFA) and/or microwave coagulation therapy (MCT) were used. Ablation therapies were performed in percutaneous, endoscopic, and operative procedures. The regional recurrence rate at the therapeutic sites was 15% (median follow-up period of 2.5 years). The average surgical margin in the operative ablation group was 11 mm. The cumulative 5-year survival rates were 37% in the local ablation, 41% in the hepatic resection, and 5% in the regional chemotherapy groups. Major complications occurred in only two patients (one biliary fistula and one liver abscess). Together these observations indicate that local ablation therapy is a radical and safe locoregional therapy that provides adequate local control and contributes to long survival. PMID- 11394003 TI - [Results and limitations of arterial infusion therapy for liver metastases from colorectal cancer]. AB - In our institution, patients with multiple unresectable liver metastases from colorectal cancer have received 24-h continuous arterial infusion therapy of 5 fluorouracil (5-FU) 250 mg/body/day every 2 weeks for the past 11 years. Although the 5-year survival rate of those who underwent surgical resection was 35%, that of unresectable patients who received 5-FU arterial infusion was 24% at 2 years, 9% at 3 years, and 50% survived a median of 391 days. Furthermore, we were able to perform surgical resection of metastatic residual lesions in patients who responded to 5-FU arterial infusion. This therapy has proven so effective that the 3-year survival rate of these patients is now 38%, and complete cures are being achieved. PMID- 11394004 TI - [Multidisciplinary treatment for liver metastasis using cytokines]. AB - The liver is an immunologic organ with liver-associated macrophages, so-called Kupffer cells, and natural killer-like primitive T cells. These cells may play an important role in resistance to liver metastasis. T cells are activated by the T cell growth factor, interleukin-2 (IL-2). Based on the theoretical rationale, a pilot study was conducted in 20 patients with liver metastases from primary colorectal cancer who underwent potentially curative liver resection, followed by adjuvant immunochemotherapy. The regimen consisted of a weekly hepatic arterial infusion of IL-2 1.4-2.1 x 10(6) units, and 5-fluorouracil 250 mg, and a bolus of mitomycin C 2-4 mg for 6 months. Of the 20 patients, 14 are still alive with a median postoperative survival of 69.7 months (September, 2000). The 5-year overall survival rate is 78%. Although recurrent cancer developed in 6 of the 20 patients, no patients had recurrence in the residual liver. We conclude that IL-2 based immunochemotherapy is useful in combination with liver resection for the prevention of liver recurrence in colorectal cancer patients with liver metastases. A multicenter, randomized trial is recommended. PMID- 11394005 TI - [Liver transplantation for liver metastases of colorectal cancer]. AB - Because of the critical shortage of donor organs, liver transplantation for metastatic tumors should be reserved for patients in whom long-term survival is possible. Transplantation in patients with metastatic tumors from colorectal cancer has resulted only in short disease-free intervals, and palliation was achieved in only a few patients. In patients with symptomatic neuroendocrine tumors unresponsive to conventional therapy, on the other hand, reasonably good disease-free intervals and prolonged survival may justify liver transplantation. PMID- 11394006 TI - [Future scope for gene therapy for liver metastasis of colon cancer]. AB - Liver metastasis is the most serious event for physicians and surgeons treating patients with colorectal cancer. Gene therapy is expected to become a novel strategy to prevent liver metastasis. Four types of clinical studies are currently underway: 1) suicide-gene therapy with the cytosine deaminase gene; 2) immune gene therapy with cytokine (inter leukin-2) or major histocompatibility complex class I gene HLA-B7; 3) tumor suppressor gene p53 therapy; and 4) lysis of p53 mutant cancer cells with E1B55k-deleted adenovirus (Onyx-015). Basic research provided several candidates for the liver metastasis-associated genes, including MMP7, DCC, CDC25B, E-cadherin, CD44, vascular endothelial growth factor, etc. There is an alternative approach to liver metastasis, which attempts to introduce a specific gene such as cytosine deaminase and TIMP-2 into the hepatocytes but not into the tumor itself. This concept is based on results showing that hepatocytes can incorporate genes more readily than cancer cells can. Recently, mutant virus therapy has been developed, which includes Onyx-015, adenovirus dl922-947, and mutant-type herpes simplex virus. These mutant types of virus specifically proliferate in the cancer cells and result in their lysis. In the future, development of gene delivery systems that are powerful and specific to cancer type is essential. PMID- 11394007 TI - [Extremely high level of serum thyroglobulin after surgery in a patient with follicular thyroid carcinoma. Recurrence, distant metastasis, or other etiology?]. AB - A 67-year-old female patient with a tumor of the thyroid underwent right lobectomy of the thyroid. The tumor was histologically diagnosed as a follicular carcinoma of the thyroid. Serum studies revealed very high (> 8,000 ng/ml) levels of thyroglobulin after surgery. We suspected distant metastases from follicular carcinoma or recurrence in the left lobe of the thyroid. She therefore underwent left lobectomy of the thyroid. No recurrence of follicular carcinoma was recognized in the resected thyroid tissue, but severe autoimmune thyroiditis was diagnosed histologically. Total-body 131I scintigraphy did not show abnormal distant uptake. Serum studies revealed very low levels of thyroglobulin (< 2.0 ng/ml) after the second surgery. We speculate that silent thyroiditis might have occurred after the first surgery, resulting in the high levels of serum thyroglobulin. Silent thyroiditis should be considered as a possible cause of high serum thyroglobulin levels. PMID- 11394008 TI - [KRN7000 inhibit hepatic metastasis of pancreatic cancer]. PMID- 11394009 TI - Vertical profiles of temperature and contaminant concentration in rooms ventilated by displacement with heat loss through room envelopes. AB - The purpose of this study is to examine the effect of heat loss through walls upon the gradients of temperature and contaminant concentration in room with displacement ventilation. It is known that conduction heat loss is governed by outside temperature, heat load inside the room, supply air temperature and overall heat transfer coefficient of walls. Experiments were conducted to measure the temperature gradient and the ventilation efficiency in the room ventilated by displacement ventilation with various combinations of heat load and temperature difference between supply air and outside air. In order to simulate the change of seasons, the supply air temperature was changed instead of the outside air temperature. The effect of supply air temperature and heat generation inside the room on the temperature gradient and the concentration of tracer gas were investigated through the experiments. As a result, it turned out that the higher the heat generation rate and the lower the supply temperature, the stronger the temperature stratification and the lower the concentration in the lower zone. Additionally, ventilation heat loss turned out to be a good index for assessing the concentration in the lower zone. Temperature differences of around 3 degrees C between supply air temperature and exhaust temperature are at least needed for displacement ventilation under the conditions of the experiment presented in this paper. PMID- 11394010 TI - Experimental validation of a computational fluid dynamics model for IAQ applications in ice rink arenas. AB - Many ice rink arenas have ice resurfacing equipment that uses fossil fuel as power. The combustion byproducts are a major source of contamination. Ventilation along with other pollution source control measures is the most widely applied strategy to lower the contaminant level below the threshold limit and maintain acceptable indoor air quality (IAQ). A computational fluid dynamics (CFD) model has been developed and used to predict the contaminant concentrations, air velocity, and air temperature distributions in ice rinks. The numerical results agree reasonably with the corresponding experimental data for both steady-state and transient conditions. The CFD model is a useful and inexpensive tool to investigate ventilation parameters, such as air distribution methods, ventilation effectiveness, air exchange rates, and various ventilation control strategies. PMID- 11394011 TI - Irritants and allergens at school in relation to furnishings and cleaning. AB - In order to study the influence of furnishings and cleaning on the indoor air quality at school, 181 randomly chosen classrooms were investigated. The amounts of open shelves, textiles and other fittings were noted, data were gathered on cleaning routines, and a number of pollutants were measured in the classrooms. In classrooms with more fabrics there was more settled dust and the concentration of formaldehyde was higher. Classrooms with more open shelves had more formaldehyde, and more pet allergens in settled dust, and classrooms with a white board, instead of a chalk board, were less dusty. Classrooms mainly cleaned through wet mopping had more airborne viable bacteria but less settled dust than classrooms mainly cleaned by dry methods. In rooms where the desks and curtains were more often cleaned, the concentrations of cat and dog allergen in settled dust were lower. It is concluded that furnishings and textiles in the classroom act as significant reservoirs of irritants and allergens and have an impact on the indoor air quality at school. PMID- 11394012 TI - Evaluation of a procedure to isolate culturable microorganisms from carpet dust. AB - Details of a method to isolate culturable bacteria and fungi from carpet dust were evaluated to isolate the greatest numbers of these agents. Four broad groups were evaluated: mesophilic and thermophilic bacteria and moderately hydrophilic and xerophilic fungi. Features studied included: 1) mixing time; 2) addition of glass beads; 3) length of time dust settled before suspensions were plated; 4) relative concentrations of microorganisms in the fibrous and fine dust fractions; and 5) storage temperature and period. The findings are preliminary because of the small number of samples, but a votexing time of at least 2 min with glass beads gave the best results in terms of the highest estimate of the concentration of culturable bacteria or fungi. Microorganisms were retrievable-from the upper portion of a suspending liquid for at least 30 min with little detectable change in concentration. Both bacteria and fungi were more abundant in the fine than the fibrous dust fraction. No significant losses were observed for samples stored up to 25 days at 5 degrees C or 25 degrees C. This procedure optimized measurement of total concentration, but may not identify the range of genera and species in dust if microorganisms present in small numbers and as single units are underrepresented relative to those in clusters and aggregates. PMID- 11394013 TI - Issues identical or closely related to what you read in Indoor Air. PMID- 11394014 TI - Dampness in buildings and health. Nordic interdisciplinary review of the scientific evidence on associations between exposure to "dampness" in buildings and health effects (NORDDAMP). AB - Several epidemiological investigations concerning indoor environments have indicated that "dampness" in buildings is associated to health effects such as respiratory symptoms, asthma and allergy. The aim of the present interdisciplinary review is to evaluate this association as shown in the epidemiological literature. A literature search identified 590 peer-reviewed articles of which 61 have been the foundation for this review. The review shows that "dampness" in buildings appears to increase the risk for health effects in the airways, such as cough, wheeze and asthma. Relative risks are in the range of OR 1.4-2.2. There also seems to be an association between "dampness" and other symptoms such as tiredness, headache and airways infections. It is concluded that the evidence for a causal association between "dampness" and health effects is strong. However, the mechanisms are unknown. Several definitions of dampness have been used in the studies, but all seems to be associated with health problems. Sensitisation to mites may be one but obviously not the only mechanism. Even if the mechanisms are unknown, there is sufficient evidence to take preventive measures against dampness in buildings. PMID- 11394015 TI - A review: fungal exposure assessment in indoor environments. AB - While the fungal exposure assessment was based on the determination of fungal propagules for a long time, recent progress has led to the development of methodology for other fungal agents, e.g. the fungal cell wall components, metabolites, and allergens, that may be responsible for health effects caused by fungal exposure. This review includes a summary of the sampling techniques and analytical methods that are currently used or are in progress for the fungal exposure assessment. Prospects for the future trends are also discussed. In the future, the development will focus on sampling techniques that allow longer sampling times, a higher sampling efficiency for relevant particle sizes, and better possibilities for a wide range of analyses. In addition, new or modified methodology based on chemical, immunochemical, and molecular biological techniques to measure fungal agents related to health effects will improve the understanding of biological responses caused by fungal exposure. PMID- 11394016 TI - Review of methods to collect settled dust and isolate culturable microorganisms. AB - Examination of settled dust is often included in investigations of indoor environments to identify the types and concentrations of particles to which building occupants may be exposed. Fungi and bacteria are among the many components in dust that have been studied. Isolation by culture is an established method that is used widely to quantify and identify microorganisms in environmental samples. However, no standard procedures for culturing fungi or bacteria from dust have been adopted widely to ensure the validity of comparing findings from different studies. This paper reviews methods various researchers have used to study surface particles and to isolate culturable microorganisms from dust. Factors that were found to differ included the method of sample collection, the ways dust was prepared for inoculation onto growth media, and the culture media chosen for specific categories of agents. The need for reference methods in environmental microbiology for use in the assessment of indoor environmental quality is discussed. PMID- 11394017 TI - [Amperometric detection in the determination of drugs by non-separating flow methods--flow injection analysis and sequential injection analysis]. AB - The present review dealing with the use of various electrochemical detection systems in flow methods of analysis of drugs (such as FIA and SIA techniques). The review covers the period of 1988 to 1998 and involves 78 references. The drugs determined are arranged according to the functional groups undergoing electrochemical transformation; for all the analytes data on the detection conditions, detection limits and ranges of quantitation are included. Advantages and drawbacks of amperometric detection of drugs in flow systems are discussed. PMID- 11394018 TI - [Sorbents for solid phase extraction and their use]. AB - Extraction on the solid phase is a rapid, simple, effective, and flexible method of preparation of samples for the analysis proper and it can be used in many areas of analysis (pharmacy, biochemistry, food industry, environmental examination, etc.). Possible use in on-line and automatic systems is also advantageous. A very wide scale of proposed sorbents and the use of their combinations thus make extraction on the solid phase an effective method for the isolation and preconcentration of the analyte from the complex matrix of the sample. PMID- 11394019 TI - [The Yemen health service and possibilities of cooperation with the Czech Republic in the area of pharmacy]. AB - The pharmaceutical market in Yemen represents a potential target for Czech pharmaceutical firms. It requires a knowledge of specific conditions of Yemeni health service and pharmacy. With regard to different cultures and languages, Yemeni pharmaceutical market is practically unknown to us. On the basis of an analysis of the materials of the Ministry of Statistics and the Ministry of Public Health, the paper presents the data about the growth in the number of inhabitants, the number of health service establishments, the structure and number of workers in health services, morbidity of the population, and a list of typical diseases. The hitherto unpublished data about inland pharmaceutical industry and import of drugs, conditions for registration of pharmaceutical firms and registration of drugs, and fees for the registration make it possible for Czech pharmaceutical firms to carry out their own marketing analyses and improve their possible activities in Yemen. PMID- 11394020 TI - [Effect of n-decane and n-octadecane on autoperoxidation of egg yolk phosphatidylcholine in multilamellar liposomes: correlation with lipid bilayer thickness and its stability]. AB - Conjugated dienes (CD) and thiobarbituric acid reactive substances (TBARS) have been estimated during the autoperoxidation of chromatographically pure phosphatidylcholine from hen eggs (EYPC) in multilamellar liposomes by UV-VIS spectrophotometry. During the propagation phase of autoperoxidation reaction, n decane (C10) and n-octadecane (C18) gradually inhibit EYPC peroxidation up to 0:2:1 and 1:1 alkane:EYPC molar ratios, respectively. At higher molar ratios, the yield of estimated autoperoxidation products increases. At the highest molar ratio studied (alkane:EYPC = 2:1), the CD and TBARS concentrations exceed their levels in control sample without alkane added. The changes in the lipid bilayer thickness estimated from the small-angle neutron scattering (SANS) curves of unilamellar dioleoylphosphatidylcholine (DOPC) liposomes have indicated that C10 is located in the bilayer hydrophobic region parallel to the DOPC acyl chains at low molar ratios (C10:DOPC < or = 0.4). At higher molar ratios (0.6 < or = C10:DOPC < or = 1.0), the alkane changes its location into the center of the bilayer between the apposing monolayers. The alkane location parallel to polyunsaturated lipid fatty acyl chains RH separates these chains resulting in a decreased frequency of their encounters, in decreased yields of ROO. + RH-->ROOH + R., 2 ROOH-->RO. + ROO. + H2O and RO. + RH-->R. + ROH free radical reactions, and consequently, in decreased autoperoxidation. Autoperoxidation returns to the control values due to alkane redistribution into the bilayer center. 1H decoupled 31P NMR spectra of EYPC + C10 aqueous dispersions have shown that the lipid bilayer transforms into nonbilayer phases at C10:EYPC > 1:1 molar ratios. The inverted hexagonal HII phase with C10 (or C18) preferentially located in the interstitial regions of the HII unit cell displays higher autoperoxidation than the control bilayer sample due to greater motional freedom of RH chains in the HII phase. PMID- 11394021 TI - [Use of over-the-counter drugs containing ibuprofen in self-medication]. AB - Ibuprofen (MO1AE01) is a suitable means for self-medication with regard to its relatively wide spectrum of indication, good tolerance, and safety. In the Czech Republic, OTC preparations containing ibuprofen represent frequently used medicaments. The paper examines solid divided dosage forms for oral administration. A survey based on questionnaires reveals in what indications and according to what dosing schemes the purchased preparations are used, and the paper evaluates possible risks. Strengthening of the role of the pharmacist as the provider of information on drugs, keeping patients' drug records in pharmacies, and deepening of the mutual cooperation of the physician and pharmacist would contribute to increase the safety of the use of the preparations under study. PMID- 11394022 TI - [Opinions and attitudes of patients visiting pharmacies]. AB - The paper is devoted to the problems concerning the opinions and attitudes of customers of pharmacies, i.e. their opinions on and attitudes to the question of information needs and satisfaction with information and services provided in pharmacies in the Czech Republic. The article presents the results of a questionnaire survey answering the following questions: why the respondents come to pharmacies and what the reason of the present visit is, whom they prefer as the dispenser of the drug and why, how often they went to a pharmacy in the recent period, from where they get information concerning drugs, whether it is advantageous for them to obtain information in a pharmacy and what information they received during the present visit, who provided the information or advice, how many pieces of information they got and how they are satisfied with them, whether they required information in a pharmacy, or whether they came to a pharmacy for this purpose, why the respondents have purposefully selected a particular pharmacy, whether they take drugs daily and which ones, what their opinion on the amount the supplementary payment is, what OTC drugs they buy, what drugs they prefer, whether they concentrate on a certain preparation, or possibly which one, what is their opinion about the scale of the assortment of OTC drugs, whether they use homeopathic medicines, and what they miss in pharmacies. PMID- 11394023 TI - [Antimicrobial salicylanilides and 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-diones]. AB - Salicylanilides and 3-phenyl-2H-1,3-benzoxazine-2,4(3H)-diones are strong antimycobacterial substances which can be considered to be potential antituberculotics. In order to be able to verify the prognostics of the relationships between the structure and antimycobacterial activity, the series of previously evaluated substances was extended to include 4' ethoxycarbonylsalicylanilide, 4'-trifluoromethylsalicylanilide, 4' cyanidosalicylanilide, 4'-thiocarbamoylsalicylanilide, 3-(4-ethoxycarbonylphenyl) 2H-1,3-benzoxazine-2,4(3H)-dione, 3-(4-trifluoromethylphenyl)-2H-1,3-benzoxazine 2,4(3H)-dione, and 3-(4-cyanidophenyl)-2H-1,3-benzoxazine-2,4-(3H)-dione. The substances were evaluated against Mycobacterium tuberculosis, M. kansasii, and M. avium. In harmony with the previous study (see ref. 1), antimycobacterial activity increased with increasing lipophilicity and electron-acceptor properties of substituents. As the values of regression coefficients were not substantially changed after the complementation of the group, the present authors consider the problem under study to be solved. PMID- 11394024 TI - [Local anesthetics. Part 154: study of micellization of homologs of heptacain chloride]. AB - We have studied the critical micellar concentration (c.m.c.) of the homologs of the local anesthetic heptacaine in the medium 0.1 mol/l NaCl, employing spectrophotometry in the UV region of the spectrum at T = 21 degrees C and pH = 4.5-5.0. On the basis of our experimental results, it is assumed that in the homologous series of amphiphilic compounds the quasi-parabolic dependence of biological potencies on the length of their hydrophobic substituent ("cut-off" effect) could be caused by c.m.c. PMID- 11394025 TI - Anorectal anatomy and physiology. AB - Knowledge of anorectal anatomy is essential for understanding the normal function of the anorectum. Its physiology is more complex, however. The maintenance of continence depends on several factors, and today clinicians have a better understanding of the usual sequence of events that leads to defecation. The extensive number of investigative techniques that have evolved have permitted better understanding of the disorders of the anal sphincters, rectum, and pelvic floor. PMID- 11394026 TI - Fecal incontinence. AB - Fecal incontinence is common and socially disabling. Only a few patients with fecal incontinence present to medical practitioners. Investigative techniques have improved, and it is possible now to define accurately functional or anatomic deficits. Careful planning of treatment with the possibility of using a variety of treatment modalities is essential. Novel conservative and surgical techniques have the potential to improve the outcome for patients with fecal incontinence. PMID- 11394027 TI - Surgical treatment of constipation and fecal incontinence. AB - Fecal incontinence is a disabling and distressing condition. Many patients are reluctant to discuss the condition with a physician. A thorough history, good physical examination, and detailed anorectal physiologic investigations can help in the therapeutic decision-making algorithm. Patients with isolated anterior sphincter defects are candidates for overlapping repair. In the presence of unilateral or bilateral pudendal neuropathy, the patient should be counseled preoperatively regarding a [table: see text] lower anticipation of success. If the injury occurred shortly before the planned surgery and neuropathy is present, it may be prudent to wait because neuropathy sometimes can resolve within 6 to 24 months of the injury. Pudendal nerve study may help determine surgical timing. An anterior sphincter defect combined with a rectovaginal fistula can be approached by overlapping sphincter repair and a concomitant transanal advancement flap. Patients who had undergone multiple such procedures may benefit from concomitant fecal diversion at the time of repeat sphincter repair. Patients with global or multifocal sphincter injury may be candidates for a neosphincter procedure. The stimulated graciloplasty and artificial bowel sphincter are reasonable options. In the absence of the availability of these techniques or because of financial constraints, consideration could be given to bilateral gluteoplasty or unilateral or bilateral nonstimulated graciloplasty. The postanal repair still serves a role in patients with isolated decreased resting pressures with or without neuropathy or external sphincter injury with minimal degrees of incontinence. Biofeedback and the Procon device may play a role in these patients. Lastly, fecal diversion must be considered as a means of improving the quality of life because the patient can participate in the activities of daily living without the fear of fecal incontinence. PMID- 11394028 TI - Manometric assessment of anorectal function. AB - A diagnostic test is useful if it can provide information regarding the underlying pathophysiology, confirm a clinical suspicion, or guide clinical management. In a prospective study, anorectal manometry was shown not only to confirm a clinical impression, but also to provide new information that was not detected clinically. The information obtained from these studies influenced the management and outcome of patients with defecation disorders (Table 1). These findings have been confirmed further by another study that showed colorectal physiologic tests provided a definitive diagnosis in 75% of patients with constipation, 66% of patients with incontinence, and 42% of patients with intractable anorectal pain. A systematic and careful appraisal of anorectal function can provide invaluable information that can guide treatment of patients with anorectal disorders. A more uniform method of performing these tests and interpreting the results is needed to facilitate a wider use of this technology for the assessment of patients with anorectal disorders. PMID- 11394029 TI - Anal Fissure. AB - Anal fissure is a common condition, and although most are short-lived and heal spontaneously, those that persist and require intervention cause considerable morbidity in an otherwise healthy young population. Traditionally, lateral internal sphincterotomy was the gold standard treatment for chronic fissures, but this procedure is associated with a risk of incontinence to some degree in 30% of patients. The discovery of pharmacologic agents that effectively cause a chemical sphincterotomy and heal most fissures has led to approximately two thirds of patients avoiding surgery. Topical 0.2% GTN ointment probably is the most widely used first-line treatment. Other drugs currently under investigation may offer effective treatment with fewer side effects. Another advantage of these novel treatments is that by acting through different pathways, they may be effective in the 30% of cases in which GTN fails, the risks associated with surgery may be avoided. Studies of botulinum toxin injection into the anal sphincter have reported excellent healing rates, although the procedure is more invasive, and patients may find it uncomfortable and less tolerable. Chemical sphincterotomy is particularly suitable in patients with associated inflammatory bowel disease, in whom sphincterotomy for anal fissure generally is contraindicated. When pharmacologic therapy fails or fissures recur frequently and patients have raised resting anal pressure, lateral internal sphincterotomy is the surgical treatment of choice. The results are satisfactory when patients are selected carefully and the incision is limited to the length of the fissure. When chemical sphincterotomy fails and resting anal pressures are not elevated, as is commonly the case with patients developing fissures postpartum, an advancement flap should be considered. PMID- 11394030 TI - Hemorrhoids. AB - Understanding of hemorrhoidal pathology and treatment has come a long way. The theory of a sliding anal canal lining and the knowledge that hemorrhoidal cushions are a normal part of the anal anatomy should encourage symptom control rather than radical removal of tissue. Techniques that fix the cushions back in position can be performed in outpatients with reasonable success rates. When required, surgery should be aimed at symptomatic hemorrhoids. It is hoped that new developments such as circular stapling and better pain management will promote increased day surgery, better pain control, and less time off work for patients. PMID- 11394031 TI - Rectal prolapse, rectal intussusception, rectocele, and solitary rectal ulcer syndrome. AB - Rectal prolapse can be diagnosed easily by having the patient strain as if to defecate. A laparoscopic rectopexy should be recommended. Intussusception is more an epiphenomenon than a cause of defecatory disorder and should be managed conservatively. Solitary rectal ulcer syndrome is a consequence of chronic straining, and therapy should include restoring a normal defecation habit. Rectocele should be left alone; an operation may be considered if it is larger than 3 cm and is causing profound symptoms despite maximizing medical therapy for the associated defecation disorder. PMID- 11394032 TI - Pouch and pouchitis. AB - Pouchitis is an inflammation of unknown origin occurring in the ileal pouch after IPAA. It is considered by many to be a form of ulcerative colitis that recurs in the pouch and rarely, if ever, occurs in patients with FAP. Most patients respond to a short course of antibiotics. When remission cannot be maintained or the disease is nonresponsive to prolonged treatment with antibiotics, anti inflammatory agents or steroids may be useful. A variety of alternative drugs have been tried with mixed success and should be considered as experimental. Rarely, when pouchitis is refractory to medical management, excision of the pouch may be required. PMID- 11394033 TI - Functional anorectal and pelvic pain. AB - Functional anorectal and pelvic pain syndromes represent a diverse group of disorders that affect the quality of life and about which many physicians possess little understanding. Nongynecologic causes include levator ani syndrome, proctalgia fugax, and coccygodnia, which can often be distinguished by careful history and physical examination. In women, chronic pelvic pain may arise from the uterus, cervix, ovaries, or from endometriosis and pelvic adhesions. This article reviews these diverse disorders and the approach to diagnosis and management. PMID- 11394034 TI - Neurologic disorders affecting the anorectum. AB - Neurologic disorders that affect the brain, spinal cord, or extrinsic innervation may present with similar symptoms and share common pathophysiology, such as rectal impaction, loss of an urge to defecate, inability to trigger a defecation sequence, obstructive defecation, or incontinence. If these symptoms are persistent or bothersome, they require treatment. The management of a patient with neurologic anorectal dysfunction depends on the underlying pathophysiologic mechanisms. Dietary advice, bowel training, pharmacotherapy, and rehabilitative treatment may be used alone or in combination. PMID- 11394035 TI - Pediatric anorectal disorders. AB - Anorectal disorders are common in children. It has been estimated that 10% of children are brought to medical attention because of a defecation disorder, and constipation is the chief complaint in 3% to 5% of all visits to pediatricians. Most of these children suffer from functional constipation. There are substantial differences in the approach to children and adults with anorectal disorders. This article discusses the commonest pediatric anorectal disorders, emphasizing the differences between children and adults in terms of clinical presentations, pathophysiology, diagnosis, and treatment. PMID- 11394036 TI - Clinical neurophysiology and electrodiagnostic testing of the pelvic floor. AB - This article summarizes our current understanding of the neuroanatomy and neurophysiology of the pelvic floor. The electrodiagnostic evaluation of the pelvic floor muscles and external anal sphincter, including pudendal nerve conduction studies, sacral reflexes, and needs EMG is presented. The discussion reviews the test methodology, the strengths and limitations of each test, and their clinical utility. The authors have tried to critically review the objective evidence to support the use of electrodiagnostic tests in the evaluation and management of pelvic floor disorders. The reader will have a better understanding of the rationale, methodology, clinical utility, and potential pitfalls for each of the commonly used neurophysiological tests of the pelvic floor. PMID- 11394037 TI - Radiologic evaluation of anorectal disorders. AB - Defecation is a dynamic event, and although evacuation proctography does not simulate physiologic defecation exactly, it does provide maximal stress to the pelvic floor and image rectal emptying, both of which are required for the diagnosis of certain conditions: MR imaging studies are attractive in that no ionizing radiation is involved, but unless an evacuation study is performed, the features of anismus, trapping in a rectocele, and intussusception cannot be diagnosed. Because these are the main reasons for investigating difficult defecation, the fluoroscopic examination is the simplest and most reliable method. Endoanal ultrasound is an ideal screening examination for incontinence to show internal sphincter degeneration and tears of the internal or external sphincters. The diagnosis of external sphincter atrophy on ultrasound is not yet resolved, and this remains an important indication for endoanal MR imaging. PMID- 11394038 TI - Slow transit constipation. AB - Slow transit constipation is a clinical syndrome predominantly affecting young women, characterized by constipation and delayed colonic transit, occasionally associated with pelvic floor dysfunction. The disorder spans a spectrum of variable severity, ranging from patients who have relatively mild delays in transit but who are otherwise indistinguishable from irritable bowel syndrome patients at one extreme, to patients with colonic inertia or chronic megacolon at the other extreme. Potential mechanisms for impaired colonic propulsion include fewer colonic HAPCs or a reduced colonic contractile response to a meal. The cause of the syndrome is unclear. The treatment is primarily medical; surgery is reserved for patients with severe disease or colonic inertia. Recognition and treatment of pelvic floor dysfunction is crucial for patients treated medically or surgically. Collaborative studies are necessary to determine the pathophysiology of this disorder and to ascertain the efficacy of novel prokinetic agents. PMID- 11394039 TI - Dyssynergic defecation. AB - Dyssynergic defecation is a common clinical problem that affects half of patients with chronic constipation. In many patients, there is a significant overlap with slow transit constipation. The chief underlying pathophysiologic mechanism is a failure of rectoanal coordination. By using a combination of history, prospective stool diaries, detailed clinical evaluation, and anorectal physiologic tests, it is possible to diagnose this problem. Controlled trials are under way to evaluate the efficacy of biofeedback therapy. Meanwhile, it is possible to treat most patients by using neuromuscular conditioning and biofeedback therapy. Further refinements in diagnostic criteria and in rehabilitation therapy programs should facilitate better diagnosis and treatment of patients with dyssynergic defecation. PMID- 11394040 TI - Growth factor regulation of kinases. PMID- 11394041 TI - The Berlin "protein structure factory" initiative: a technology-oriented approach to structural genomics. PMID- 11394042 TI - Structure-based approaches in modern drug discovery research. PMID- 11394043 TI - DNA recognition by NF kappa B and STAT transcription factors. PMID- 11394044 TI - Molecular determinants for agonist and antagonist binding to steroid nuclear receptors. PMID- 11394045 TI - Estrogen receptor-cofactor interactions as targets for novel drug discovery. PMID- 11394046 TI - Cell cycle regulation by protein kinases and phosphatases. PMID- 11394047 TI - Structural basis for substrate recognition and control in protein kinases. PMID- 11394048 TI - The selectivity of small molecules towards protein tyrosine kinases. PMID- 11394049 TI - Elements of signal transduction in drug discovery with special reference to inhibitors of protein kinase C. PMID- 11394050 TI - Does negative priming reflect inhibitory mechanisms? A review and integration of conflicting views. AB - Negative priming has traditionally been viewed as a reflection of an inhibitory mechanism of attention. However, recent accounts have suggested that negative priming does not reflect inhibitory mechanisms. Rather, slowed reaction times on negative priming trials are either due to retrieval of incompatible response tags or of mismatching perceptual information, or due to extra processes needed to distinguish past from present information. In contrast, it is proposed that there is no firm evidence to discount inhibition models. In fact, although retrieval processes can be implicated in negative priming effects, understanding of these requires consideration of the inhibitory processes involved in selecting information for goal-directed behaviour. PMID- 11394051 TI - Implicit sequence learning with competing explicit cues. AB - Previous research has shown that the expression of implicit sequence learning is eliminated in a choice reaction time task when an explicit cue allows participants to accurately predict the next stimulus (Cleeremans, 1997), but that two contingencies predicting the same outcome can be learned and expressed simultaneously when both of them remain implicit (Jimenez & Mendez, 1999). Two experiments tested the hypothesis that it is the deliberate use of explicit knowledge that produces the inhibitory effects over the expression of implicit sequence learning. However, the results of these experiments do not support this hypothesis, rather showing that implicit learning is acquired and expressed regardless of the influence of explicit knowledge. These results are interpreted as reinforcing the thesis about the automatic nature of both the acquisition and the expression of implicit sequence learning. The contradictory results reported by Cleeremans are attributed to a floor effect derived from the use of a special type of explicit cue. PMID- 11394052 TI - Priming effects from phonologically related distractors in picture-word interference. AB - In the cross-modal picture-word interference task, distractors phonologically related to a to-be-named picture facilitate the naming response as compared to unrelated distractors. Our experiment shows that this phonological priming effect can be obtained with as early an SOA as -300 ms. The experiment also demonstrates that this priming effect cannot be attributed to strategic behaviour of the participants as opposed to automatic preactivation processes in the lexical conceptual system. The implications for studies using the picture-word interference task as a tool for investigating lexicalization processes in speech production are discussed. PMID- 11394053 TI - Best face forward: similarity effects in repetition priming of face recognition. AB - Two experiments examined the graded similarity effect in the repetition priming of familiar face recognition. From the model of repetition priming proposed by Burton, Bruce, and Johnston (1990) it was predicted that similarity effects may be a confound of stimulus preparation. Experiment 1 was used to discount this hypothesis, but failed to replicate a pattern of graded priming related to the similarity of prime and target faces. Experiment 2 attempted a more extensive investigation using two different measures of prime-target similarity. The results replicated Ellis, Young, Flude, and Hay's (1987) finding that similar primes confer more priming than dissimilar ones, but found no correlation between amount of priming and the degree of prime-target resemblance for either similarity metric used. In view of these findings the mechanism of repetition priming in familiar face recognition is discussed. PMID- 11394054 TI - Development of memory for pattern and path: further evidence for the fractionation of visuo-spatial memory. AB - Evidence from a number of sources now suggests that the visuo-spatial sketchpad (VSSP) of working memory may be composed of two subsystems: one for maintaining visual information and the other for spatial information. In this paper we present three experiments that examine this fractionation using a developmental approach. In Experiment 1, 5-, 8-, and 10-year old children were presented with a visuo-spatial working memory task (the matrices task) with two presentation formats (static and dynamic). A developmental dissociation in performance was found for the static and dynamic conditions of both tasks, suggesting that the activation of separable subsystems of the VSSP is dependent upon a static/dynamic distinction in information content rather than a visual/spatial one. A highly similar pattern of performance was found for a mazes task with static and dynamic formats. However, one strategic activity, the use of simple verbal recoding, may also have been responsible for the observed pattern of performance in the matrices task. In Experiments 2 and 3 this was investigated using concurrent articulatory suppression. No evidence to support this notion was found, and it is therefore proposed that static and dynamic visuo-spatial information is maintained in working memory by separable subcomponents of the VSSP. PMID- 11394055 TI - Syllabic organization and deafness: orthographic structure or letter frequency in reading? AB - For hearing people, structure given to orthographic information may be influenced by phonological structures that develop with experience of spoken language. In this study we examine whether profoundly deaf individuals structure orthographic representation differently. We ask "Would deaf students who are advanced readers show effects of syllable structure despite their altered experience of spoken language, or would they, because of reduced influence from speech, organize their orthographic knowledge according to groupings defined by letter frequency?" We used a task introduced by Prinzmetal (Prinzmetal, Treiman, & Rho, 1986) in which participants were asked to judge the colour of letters in briefly presented words. As with hearing participants, the number of errors made by deaf participants was influenced by syllable structure (Prinzmetal et al., 1986; Rapp, 1992). This effect could not be accounted for by letter frequency. Furthermore, there was no correlation between the strength of syllable effects and residual speech or hearing. Our results support the view that the syllable is a unit of linguistic organization that is abstract enough to apply to both spoken and written language. PMID- 11394056 TI - Neuropsychological evidence for case-specific reading: multi-letter units in visual word recognition. AB - We describe a patient (GK) who shows symptoms associated with Balint's syndrome and attentional dyslexia. GK was able to read words, but not nonwords. He also made many misidentification and mislocation errors when reporting letters in words, suggesting that his word-naming ability did not depend upon preserved position-coded, letter identification. We show that GK was able to read lower case words better than upper-case words, but upper-case abbreviations better than lower-case abbreviations. Spacing the letters in abbreviations disrupted identification, as did mixing the case of letters within words. These data cannot be explained in terms of letter-based reading or preserved holistic word recognition. We propose that GK was sensitive to the visual familiarity of adjacent letter forms. PMID- 11394057 TI - Age of acquisition and word frequency in written picture naming. AB - This study investigates age of acquisition (AoA) and word frequency effects in both spoken and written picture naming. In the first two experiments, reliable AoA effects on object naming speed, with objective word frequency controlled for, were found in both spoken (Experiment 1) and written picture naming (Experiment 2). In contrast, no reliable objective word frequency effects were observed on naming speed, with AoA controlled for, in either spoken (Experiment 3) or written (Experiment 4) picture naming. The implications of the findings for written picture naming are briefly discussed. PMID- 11394058 TI - Task-switching costs, Stroop-costs, and executive control: a correlational study. AB - Three correlational studies investigated the relationship between the time costs associated with Stroop stimuli (Stroop-costs) with the time costs associated with task-switching (switch-costs) obtained from colour-word stimuli and digit stimuli. In all studies, large and significant positive correlations were found between different measures of switch-costs. However, only small (and sometimes non-significant) correlations were obtained between the different measures of Stroop-costs and between measures of Stroop-costs and measures of switch-costs. The results are taken as evidence for the existence of some common or shared specialized mechanisms involved in task-switching, which are different from those used to overcome Stroop interference. PMID- 11394060 TI - Relative importance of perceptual and mnemonic variance in human temporal bisection. AB - We investigated the relative contributions of perception and reference memory to behavioural variability in a temporal discrimination with human subjects. We used two temporal bisection tasks. In both tasks each trial consisted of a sequential presentation of three intervals, two standards, and a probe, and subjects were asked to judge the similarity of each probe against the two standards. In a "single bisection", the standards' duration was constant across trials. In a "roving bisection", the two standards were trial unique. We compared our results with the predictions from a model related to Scalar Expectancy Theory, with the added assumption that the decision process minimizes the expected number of errors given the information available. The model shows that if errors in reference memory were dominant the psychometric function should be identical for single and roving tasks, and if perceptual errors were dominant the psychometric function should be steeper for the single than for the roving bisection. As we found that psychometric functions were steeper for the single than for the roving tasks, we concluded that perceptual errors are dominant. PMID- 11394059 TI - The reproduction of vertical and oblique orientations in the visual, haptic, and somato-vestibular systems. AB - This study investigates whether the vertical orientation may be predominantly used as an amodal reference norm by the visual, haptic, and somato-vestibular perceptual systems to define oblique orientations. We examined this question by asking the same sighted adult subjects to reproduce, in the frontal (roll) plane, the vertical (0 degree) and six oblique orientations in three tasks involving different perceptual systems. In the visual task, the subjects adjusted a moveable rod so that it reproduced the orientation of a visual rod seen previously in a dark room. In the haptic task, the blindfolded sighted subjects scanned an oriented rod with one hand and reproduced its orientation, with the same hand, on a moveable response rod. In the somato-vestibular task, the blind folded sighted subjects, sitting in a rotating chair, adjusted this chair in order to reproduce the tested orientation of their own body. The results showed that similar oblique effects (unsigned angular error difference between six oblique orientations and vertical orientation) were observed across the three tasks. However, there were no positive correlations between the visual, haptic, and somato-vestibular oblique effects. Moreover, in some oblique orientations, there was a tendency to overestimate the angle between the oblique orientation and the vertical orientation. This effect varied according to the orientation value and the modality. Taken together, these findings suggest that although vertical orientation is used as a reference norm in the visual, haptic, and somato-vestibular systems to define oblique orientations, specific processing mechanisms seem to be at work in each perceptual system. PMID- 11394061 TI - Concurrent and retroactive interference effects in memory of movement patterns. AB - Three experiments examined memory for enacted and observed movement patterns. The effect of visuo-spatial tracing tasks requiring overt motor movements, comparable to those required by the to-be-recalled movement pattern task when enacted, were compared with the effects of non-motor interference tasks, such as articulatory suppression and counting backwards. None of the experiments was able to detect indices of motor mechanisms in memory of movement patterns. The results are discussed in terms of theories of action memory and working memory and of a problem-solving account. PMID- 11394062 TI - The acquisition of odour qualities. AB - Odours can evoke a large range of qualities. Some of these qualities (e.g., sweetness) appear to be acquired through simultaneously experiencing odours mixed with tastes. Acquisition might also occur when two odours are experienced as a mixture. In this case mixture components might acquire each other's qualities. This was tested in two experiments. In the first, subjects repeatedly sniffed two odour mixtures (either AX, BY or AY, BX). Odours mixed with A acquired A's properties and were judged more similar to A than to B. Odours mixed with B were not clearly discriminable. The second experiment used a similar approach except that Odour B was replaced. Subjects now smelled either AX, CY or AY, CX. All components were discriminable. Odours mixed with A acquired A's properties and were judged more similar to A than to C. Although odours mixed with C did not acquire C's qualities due to a confusion of quality terms, odours previously mixed with C were judged as more similar to C than to A. Evidence of other quality exchanges were also obtained. These results suggest that pure odour qualities can be learnt and lend support to William James's (1890) notion of the acquired equivalence/distinctiveness of cues. PMID- 11394063 TI - Mental planning and the Tower of London task. AB - The Tower of London (TOL) task has been used extensively as a test of planning ability in neuropsychological patients and normal populations. Participants are asked to preplan mentally a sequence of moves to match a start set of discs to a goal, and then to execute the moves one by one. The mental preplanning stage has been identified as critical to efficient performance. The current experiments examined whether manipulations of mental preplanning influence performance on the TOL. In Experiment 1, the effect of different planning instructions was examined. Those told to make full mental plans spent considerably longer in planning than participants given no specific planning instructions, yet there was no effect of instruction condition on the efficiency of executing plans. Experiment 2 investigated whether people were able to plan mentally, by looking at their ability to identify intermediate states of an optimum mental plan. Results indicated that most individuals could make accurate preplans up to two subgoals ahead, but not three. However, making an efficient preplan did not result in better subsequent execution of moves to solve the TOL trial. It is concluded that people can make effective mental plans for a limited number of moves. However, on the TOL task, mental preplanning does not offer benefits in terms of quicker performance, or more accurate solution. The nature of planning in the TOL task is therefore questioned. PMID- 11394064 TI - Algorithmic solution of arithmetic problems and operands-answer associations in long-term memory. AB - Many developmental models of arithmetic problem solving assume that any algorithmic solution of a given problem results in an association of the two operands and the answer in memory (Logan & Klapp, 1991; Siegler, 1996). In this experiment, adults had to perform either an operation or a comparison on the same pairs of two-digit numbers and then a recognition task. It is shown that unlike comparisons, the algorithmic solution of operations impairs the recognition of operands in adults. Thus, the postulate of a necessary and automatic storage of operands-answer associations in memory when young children solve additions by algorithmic strategies needs to be qualified. PMID- 11394065 TI - Are conjunctive inferences easier than disjunctive inferences? A comparison of rules and models. AB - We report four experiments investigating conjunctive inferences (from a conjunction and two conditional premises) and disjunctive inferences (from a disjunction and the same two conditionals). The mental model theory predicts that the conjunctive inferences, which require one model, should be easier than the disjunctive inferences, which require multiple models. Formal rule theories predict either the opposite result or no difference between the inferences. The experiments showed that the inferences were equally easy when the participants evaluated given conclusions, but that the conjunctive inferences were easier than the disjunctive inferences (1) when the participants drew their own conclusions, (2) when the conjunction and disjunction came last in the premises, (3) in the time the participants spent reading the premises and in responding to given conclusions, and (4) in their ratings of the difficulty of the inferences. The results support the model theory and demonstrate the importance of reasoners' inferential strategies. PMID- 11394066 TI - [Socioprofessional future in adulthood of children with end-stage kidney failure]. PMID- 11394067 TI - [Future in adulthood of children with diabetes]. PMID- 11394068 TI - [Outcome in adulthood of children and adolescents with chronic intestinal inflammatory diseases]. PMID- 11394069 TI - [Modalities of adulthood passage in children with mucoviscidosis]. PMID- 11394070 TI - [Adolescents in the year 2001]. PMID- 11394071 TI - [Emergencies in pediatric rheumatology]. PMID- 11394073 TI - [Role of the surgeon in the diagnosis and management of rheumatic diseases]. PMID- 11394072 TI - [Long-term corticotherapy in children: how to reduce its side effects?]. PMID- 11394074 TI - [Pediatric rheumatology: a multidisciplinary overspecialty]. PMID- 11394075 TI - [Pharmacology of antiretroviral agents in children]. PMID- 11394076 TI - [Varicella and acyclovir]. PMID- 11394077 TI - [Antiviral agents in the treatment of influenza]. PMID- 11394078 TI - [Diagnosis and management of cytomegalovirus infection in the fetus and the newborn]. PMID- 11394079 TI - [Physiopathology of acute severe asthma]. PMID- 11394080 TI - [Hospitalization criteria in emergency asthmatic+ crises]. PMID- 11394081 TI - [Viral and atypical germ infections in asthmatic crisis in children]. PMID- 11394082 TI - [Drug therapy of acute severe asthma]. PMID- 11394083 TI - [Pediatric malaria imported in France]. PMID- 11394084 TI - [Halofantrine in pediatrics: survey in Ile-de-France]. PMID- 11394085 TI - [Treatment of malaria]. PMID- 11394086 TI - [Prevention of malaria]. PMID- 11394087 TI - [Diagnostic imaging of neonatal tumors: certainties and uncertainties]. PMID- 11394088 TI - [Surgical management of malignant tumors in newborns]. PMID- 11394089 TI - [Indications for medical treatment for tumors in newborns]. PMID- 11394090 TI - [Epidemiologic bases of obesity]. PMID- 11394091 TI - [Management of beginning weight overloads. Physiopathologic bases and practical modalities]. PMID- 11394092 TI - [Management of severe obesity in adolescents]. PMID- 11394093 TI - [Ionic channels and epilepsy]. PMID- 11394094 TI - [Hyperinsulinism]. PMID- 11394095 TI - [Hereditary renal tubule diseases]. PMID- 11394096 TI - [Cystic kidney in children. What is new in 2001?]. PMID- 11394097 TI - ["New" explorations of renal function]. PMID- 11394098 TI - [Update on nephrotic syndrome]. PMID- 11394099 TI - [Dialysis in children: new findings]. PMID- 11394100 TI - ["New renal insufficiencies"]. PMID- 11394101 TI - [Contribution of molecular biology in the diagnosis of monogenic hereditary nephropathy]. PMID- 11394102 TI - [Neurotoxicity of cytostatic treatments in pediatrics]. PMID- 11394103 TI - [Hepatitis B vaccination and neurologic manifestations in children]. PMID- 11394104 TI - [Antiviral agents and mitochondrial diseases]. PMID- 11394105 TI - [Neurologic complications associated with antalgic and anti-inflammatory agents]. PMID- 11394106 TI - [Approach at the time of birth in suspected congenital adrenal hyperplasia]. PMID- 11394107 TI - [Emergence of type II diabetes in children: diagnostic problems]. PMID- 11394108 TI - [Complications of type 1 diabetes in children: state of affairs]. PMID- 11394109 TI - [Neonatal diabetes]. PMID- 11394110 TI - [Pharmacogenetics in pediatrics]. PMID- 11394111 TI - [Implication of maturation of enzymes of drug metabolism in development]. PMID- 11394112 TI - [Genetic control of maturation of drug metabolism]. PMID- 11394113 TI - [Pharmacogenetics of cytochromes P450: practical implications]. PMID- 11394114 TI - [Molecular genetics of Williams' syndrome]. PMID- 11394115 TI - [Physiopathology of Williams' syndrome arteriopathy]. PMID- 11394116 TI - [Cognitive, personality and behavioral characteristics -- social incidence]. PMID- 11394117 TI - [New cytogenetic study technique in hematology]. PMID- 11394118 TI - [Role of genetic markers in the diagnosis and prognosis of tumors in children]. PMID- 11394119 TI - [New prognostic factors of neuroblastoma]. PMID- 11394120 TI - [Children's health in developing countries]. PMID- 11394121 TI - [Respiratory diseases and sleep during development]. PMID- 11394122 TI - [Explorations of sleep in infants and children]. PMID- 11394123 TI - [Sleep apnea syndrome, from diagnosis to treatment]. PMID- 11394124 TI - [Ondine's syndrome]. PMID- 11394125 TI - [Clinical approach to the dysmorphic child]. PMID- 11394126 TI - [Genes and craniofacial dysmorphisms]. PMID- 11394127 TI - [Animal models of craniofacial dysmorphisms]. PMID- 11394128 TI - [Craniofacial development: morphogenesis and determinism]. PMID- 11394129 TI - [Looking at the dysmorphic child; the look of the dysmorphic child]. PMID- 11394130 TI - [Diagnostic tests and treatment of Helicobacter pylori gastritis in children]. PMID- 11394131 TI - [Severe intractable diarrhea in infants]. PMID- 11394132 TI - [New diagnostic strategies of celiac disease]. PMID- 11394133 TI - [New therapeutic approaches in Crohn's disease]. PMID- 11394134 TI - [Physiopathology of pancreatic lesions in mucoviscidosis]. PMID- 11394135 TI - [Neonatal cholestasis: diagnosis and etiology]. PMID- 11394136 TI - [Resuscitation in the delivery room: techniques of manual ventilation]. PMID- 11394137 TI - [Heart massage and adrenaline in the newborn at the delivery room]. PMID- 11394138 TI - [Management of a newborn in the case of meconial amniotic fluid]. PMID- 11394140 TI - [Munchausen syndrome by proxy]. PMID- 11394139 TI - [Surgical emergencies at the delivery room]. PMID- 11394141 TI - [The shaken baby syndrome]. PMID- 11394142 TI - [How to notify the authorities about child abuse]. PMID- 11394143 TI - [Management of ventricular dilatation]. PMID- 11394144 TI - [Immobility fetal syndrome. Warning signs to the etiopathogenic diagnosis]. PMID- 11394145 TI - [Role of neuropediatrics in prenatal diagnosis]. PMID- 11394146 TI - [Understanding somatic expressions of psychological distress]. PMID- 11394147 TI - [Gastroesophageal reflux: is it a somatic expression of psychologic distress in infants?]. PMID- 11394148 TI - [Gastroesophageal reflux: the psychotherapeutic approach]. PMID- 11394149 TI - [Clinical signs of psychological distress]. PMID- 11394150 TI - [Taking into account psychologic distress in general pediatrics]. PMID- 11394151 TI - [Depression in babies]. PMID- 11394152 TI - [Skin and soft tissue bacterial infections in newborns]. PMID- 11394153 TI - [Cellulitis in children]. PMID- 11394154 TI - [Severe microbial skin infections]. PMID- 11394155 TI - [Antibiotic treatment of skin infections in children]. PMID- 11394156 TI - [Do non-steroidal anti-inflammatory agents favor the occurrence of necrotizing fasciitis?]. PMID- 11394157 TI - [Adolescents in hospital emergencies: use and messages]. PMID- 11394158 TI - [Adolescents seen at the pediatrician's office in emergencies]. PMID- 11394159 TI - [Contravening adolescents: the example of the school institution]. PMID- 11394160 TI - [Sexual violence against adolescents: beyond the emergency of the medico-legal evidence]. PMID- 11394161 TI - [Reduction of the length of stay of mothers at the maternity unit: why does the pediatrician remain careful?]. PMID- 11394162 TI - [Management before delivery, screening of risk situations]. PMID- 11394163 TI - [Role of the maternity pediatrician]. PMID- 11394164 TI - [Early maternity discharge: role of the liberal pediatrician]. PMID- 11394165 TI - [The situation in the other countries of the European Community]. PMID- 11394166 TI - [Universal medical coverage: how to use it]. PMID- 11394167 TI - [Epidemiology of childhood saturnism in France]. PMID- 11394168 TI - [Saturnine fetal disease]. PMID- 11394169 TI - [Saturnism: the law and its application]. PMID- 11394170 TI - [Hereditary hypercholesterolemia: genetic causes and risks in adulthood]. PMID- 11394171 TI - [Arterial function in hypercholesterolemic children]. PMID- 11394172 TI - [Management of hypercholesterolemia]. PMID- 11394173 TI - [Smoking and lung growth]. PMID- 11394174 TI - [Passive smoking and respiratory disease]. PMID- 11394175 TI - [Why do adolescents smoke?]. PMID- 11394176 TI - [Smoking prevention]. PMID- 11394177 TI - How much has ORT reduced child mortality? PMID- 11394179 TI - Characteristics of children hospitalized with severe dehydration and persistent diarrhoea in Bangladesh. AB - The study analyzed data from a systematic sample of children, aged less than five years, who presented with persistent diarrhoea (diarrhoea of more than 14 days duration). It aims to differentiate (a) non-severe persistent diarrhoea (with no or mild dehydration) and (b) severe persistent diarrhoea (with moderate or severe dehydration), and to identify individual characteristics associated with severe persistent diarrhoea. In total, 7,505 patients, who represented a 4% systematic sample of the patient population, were seen during January 1993-December 1995. Of them, 297 (4%) presented with persistent diarrhoea. The male:female ratio was 2:1. Eighty-three percent of them had mild or no dehydration, and 17% had moderate or severe dehydration. Severe malnutrition of the study patients defined as weight-for-age z-score < -3, weight-for-length z-score < -3 and length-for-age z-score < -3 were 33.9%, 9.7%, and 22.7% respectively. Only 3% had oedematous malnutrition, and 11% had xerophthalmia. Factors independently associated with severe persistent diarrhoea by logistic regression analyses were: number of watery stool > 10 times during the last 24 hours prior to admission (OR, 10.0; CI, 1.2-87, p = 0.03), lower respiratory tract infection (OR, 111; CI, 4.2-2955, p = 0.004), and lack of mothers' education (OR, 7.8; CI, 1.4-41.9, p = 0.016) after controlling for confounders. Awareness and health education of mothers or caregivers and better case management during acute diarrhoeal episode might prevent the development of severe persistent diarrhoea in young children. In addition, children with severe persistent diarrhoea might need special attention to have adequate rehydration and control of extraintestinal infections, including respiratory tract infection. PMID- 11394178 TI - Clinical characteristics of non-insulin-dependent diabetes mellitus among southwestern American Indian youths. AB - The clinical characteristics and presentation of non-insulin-dependent diabetes mellitus (NIDDM) among 22 youths, aged less than 20 years, of an American Indian tribe Tohono O'odham Nation in the southwestern United States were studied. Ten males and 12 females (7-20 years old) were identified with a 13.7-year mean age of onset of diabetes. Over 80% (18/22) of the patients were obese at diagnosis having a body mass index greater than the 95th percentile for their age and sex, and there was a strong family history of NIDDM; eight patients were born to mothers who had gestational diabetes, and 19 patients had at least one parent with NIDDM. At the time of diagnosis, plasma glucose levels ranged from 10.3 mmol/L to 33 mmol/L, with nearly 60% (13/22) of the patients having a glucose reading greater than 16.8 mmol/L. C-peptide levels were done on 10 patients, and these were in the normal to elevated range. Clinical management of the 22 patients varied. To control hyperglycaemia and symptoms, such as nocturia and polyuria, 14 patients were on oral hypoglycaemic medication, and five were on insulin therapy. Compliance with dietary management was very difficult for these patients as evidenced by the fact that only three patients were on dietary control for their diabetes. The cases described in this series demonstrate NIDDM in childhood and illustrate the importance of accurate classification of diabetes during childhood, particularly in children from populations at high risk for NIDDM. PMID- 11394180 TI - Prevalence of viral, bacterial and parasitic enteropathogens among young children with acute diarrhoea in Jeddah, Saudi Arabia. AB - The prevalence of viral, bacterial and parasitic pathogens among children of Jeddah, Saudi Arabia, was investigated. During December 1995-October 1996, 576 faecal samples were collected from children (0-5 year(s) old) suffering from acute diarrhoea and attending hospitals and outpatient clinics in Jeddah. One or more enteropathogen(s) were identified in 45.6% of the stool specimens. Mixed infections were detected in 12.2% of the diarrhoeal cases. Rotavirus was detected in 34.6% of the specimens of the hospitalized patients and in 5.9% of the specimens of the outpatients. Fifty-one percent of the rotavirus-positive specimens were long electropherotype, 26% were short electropherotype, and 23% could not be electropherotyped specifically. Among those of the long electropherotype, there were six patterns; and of the short electropherotypes, there were four patterns. Serotyping of these specimens revealed a distribution of 39.6%, 4.2%, 6.3%, and 15.6% for rotavirus serotype 1, 2, 3, and 4 respectively. Mixed serotypes were found in 3.1%, and 31.3% of the specimens were untypeable. Other aetiologic agents recognized included Escherichia coli (13%), of which 3.8% were enteropathogenic E. coli (EPEC) and 1.9% enterohaemorrhagic E. coli. Among the E. coli (EPEC) serotypes, O111:K58:B4, O55:K59:B11, and 0127:K63:B8 were found in 31.8%, 18.2%, and 13.6% of the cases respectively. Serotype 026:K60:B6, 0124:K72:B17, and 0112:K66:B11 each was found in 9.1% of the EPEC cases. 0128:K67:B12 and 0125:K70:B13 each was found in one case only. Other detected pathogens were: Klebsiella pneumoniae (4%), Giardia lamblia (3.1%), Salmonella sp. (3%), Shigella flexneri (2.6%), Entamoeba histolytica (2.2%), Trichuris trichiura, Hymenolepis nana, and Ascaris lumbricoides (0.7% each), and Candida albicans (0.5%). Based on the results of this study, it is concluded that the high prevalence of the various enteropathogens among young children is a significant public health problem. PMID- 11394181 TI - Molecular epidemiology of Escherichia coli isolated from young South African children with diarrhoeal diseases. AB - Molecular techniques were used for studying the epidemiology of diarrhoeal infections due to Escherichia coli in the Gauteng region in South Africa. In total, 151 E. coli strains isolated from stools of patients with diarrhoea and 30 strains isolated from stools of healthy individuals were collected between March 1996 and May 1997. The E. coli isolates were characterized by serotyping, antimicrobial susceptibility testing, and adherence patterns. Polymerase chain reaction (PCR) was performed to determine the presence of the genes-encoding virulence factors. PCR showed that 59 (32.6%) of the E. coli isolates carried eaeA genes, 6 (3.3%) possessed bfpA genes, 4 (2.2%) CNF1, and 2 (1.1%) carried labile toxin and Stx2 genes. The eae genes were more prevalent in strains isolated from patients than in those from the control group (p < 0.001). Forty eight (26.5%) strains belonged to enteropathogenic E. coli (EPEC) O serogroups and 14 (7.7%) to Shiga toxin-producing E. coli (STEC) O157 serotype. A high percentage (28.2%) of atypical EPEC strains possessing the eaeA but not the bfpA genes was isolated. Most isolates were susceptible to commonly-used antimicrobial agents. The adherence of the E. coli strains to HeLa cells was identified more in patients (69.4%) than in the control group (60%) and was more dominant in infants than in adults. PCR and tissue culture assays were shown to be useful techniques for the epidemiological study of E. coli where this organism is a major cause of diarrhoea. PMID- 11394182 TI - Vibrio cholerae resistant to 2,4-diamino-6,7-diisopropylpteridine (O/129) isolated from patients with enteritis in Ceara, Brazil. AB - This paper reports the characterization of clinical Vibrio cholerae resistant to vibriostatic agent O/129, using classical and plasmid analysis. In a study conducted during December 1991-September 1993, two of 7,058 V. cholerae strains, obtained from patients suspected to have cholera in the State of Ceara, northeast Brazil, were resistant to 150 micrograms of the vibriostatic agent O/129 (2,4 diamino-6,7-diisopropylpteridine). One strain was identified as V. cholerae O1 El Tor Inaba and the other one as serogroup O22. Only one O1 strain harboured a plasmid of 147 kb transferable to Escherichia coli K12, and five strains of V. cholerae O1 and non-O1 were sensitive to O/129 and plasmid-negative at a frequency between 8 x 10(-2) and 3.6 x 10(-5). Additionally, O/129-resistant strains of V. cholerae O1 and O22 were resistant to trimethoprim/sulphamethoxazole. PMID- 11394183 TI - Developing bioethics in developing countries. PMID- 11394184 TI - Colonization of Vibrio cholerae among persons in contact with cholera patients. PMID- 11394185 TI - Health ethics in Pakistan: a literature review of its present state. AB - National literature on ethics provides an insight into the nature and development of a dialogue on health issues within a population. This study investigated the health ethics discourse in Pakistan. The purpose was to critically reflect on the nature and level of such discussions with the aim of stimulating an interest in the ethical implications of health and medicine in developing countries. The study evaluated the literature on biomedical and health ethics published in Pakistan during 1988-1999. Overall, there is a dearth of published discourse on healthcare ethics in Pakistan. Values that are considered to stem from religious teachings predominate in discussions relating to medical ethics. A lack of effective policy and legislation concerning the ethical practice of medicine is reported to have negative effects on the profession. Research ethics has not been captured in the published papers in Pakistan. Consideration of ethical issues in health is at an early stage in the country and may reflect the situation in a large part of the developing world. PMID- 11394186 TI - Metastasis of cancer: a conceptual history from antiquity to the 1990s. PMID- 11394187 TI - Dietary fat intake and the brain: a developing frontier in biological psychiatry. PMID- 11394188 TI - Unmasking social anxiety disorder. PMID- 11394189 TI - Toward an integrative understanding of social phobia. AB - Our objective was to examine the neurobiology of social phobia from the perspectives of basic sciences, genetics, immunology, neuroendocrinology, neurotransmission and neuroimaging and to provide an integrated understanding of social phobia in the framework of a hypothetical neural circuit. Family and twin studies provide evidence that social phobia is heritable with significant genetic influence, and molecular genetics offers possibilities in understanding the nature of the trait that is transmitted. The biologic distinctiveness of social phobia from anxiety disorders and physiological validation of differences between generalized and discrete social phobia subtypes have been implicated in genetic, naturalistic and chemical challenge studies. Evidence of specific dysfunction of dopaminergic, serotonergic, noradrenergic and GABAergic (gamma-aminobutyric acid) neurotransmitter systems has been presented in animal models, challenge studies and treatment investigations. Preliminary neuroimaging research supports previous studies suggesting striatal dopaminergic dysfunction in social phobia and suggests the importance of functional circuits. A neural circuit involving the striatum, thalamus, amygdala and cortical structures may provide a framework for integrating much of the current knowledge on the neurobiology of social phobia. PMID- 11394190 TI - Apomorphine and the dopamine hypothesis of schizophrenia: a dilemma? AB - The dopamine (DA) hypothesis of schizophrenia implicates an enhancement of DA function in the pathophysiology of the disorder, at least in the genesis of positive symptoms. Accordingly, apomorphine, a directly acting DA receptor agonist, should display psychotomimetic properties. A review of the literature shows little or no evidence that apomorphine, in doses that stimulate postsynaptic DA receptors, induces psychosis in non-schizophrenic subjects or a relapse or exacerbation of psychotic symptoms in patients with schizophrenia. After a detailed review of the literature reporting psychotogenic effects of apomorphine in patients with Parkinson's disease, an interpretation of these data is difficult, in part because of several confounding factors, such as the concomitant use of drugs known to induce psychosis and the advanced state of the progressive neurological disorder. In the context of the DA hypothesis of schizophrenia, the limited ability of apomorphine to induce psychosis, in contrast to indirectly acting DA agonists that increase synaptic DA, may be explained by the relatively weak affinity of apomorphine for the D3 receptor compared with DA. Alternatively, enhancement of DA function, though necessary, may be insufficient by itself to induce psychosis. PMID- 11394191 TI - Effect of the herbal extract combination Panax quinquefolium and Ginkgo biloba on attention-deficit hyperactivity disorder: a pilot study. AB - OBJECTIVE: A combination herbal product containing American ginseng extract, Panax quinquefolium, (200 mg) and Ginkgo biloba extract (50 mg) (AD-FX; CV Technologies, Edmonton, Alta.) was tested for its ability to improve the symptoms of attention-deficit hyperactivity disorder (ADHD). DESIGN: Open study. PATIENTS: 36 children ranging in age from 3 to 17 years who fit the diagnostic criteria for ADHD. INTERVENTIONS: AD-FX capsules were taken twice a day on an empty stomach for 4 weeks. Patients were instructed not to change any other medications during the study. OUTCOME MEASURES: At the beginning of the study, after 2 weeks, and then at the end of the 4-week trial, parents completed the Conners' Parent Rating Scale--revised, long version, a questionnaire that assesses a broad range of problem behaviours (and was used as an indication of ADHD symptom severity). RESULTS: After 2 weeks of treatment, the proportion of the subjects exhibiting improvement (i.e., decrease in T-score of at least 5 points) ranged from 31% for the anxious-shy attribute to 67% for the psychosomatic attribute. After 4 weeks of treatment, the proportion of subjects exhibiting improvement ranged from 44% for the social problems attribute to 74% for the Conners' ADHD index and the DSM IV hyperactive-impulsive attribute. Five (14%) of 36 subjects reported adverse events, only 2 of which were considered related to the study medication. CONCLUSIONS: These preliminary results suggest AD-FX treatment may improve symptoms of ADHD and should encourage further research on the use of ginseng and Ginkgo biloba extracts to treat ADHD symptoms. PMID- 11394192 TI - New-onset delusions and hallucinations in patients infected with HIV. AB - OBJECTIVE: To assess the relationship between HIV-associated psychotic symptoms (i.e., delusions, hallucinations) and demographic, psychopathological and medical variables by comparing patients with and without cerebral opportunistic infections or metabolic encephalopathy. DESIGN: Cross-sectional study. PATIENTS: 26 patients admitted to hospital with HIV and new-onset psychotic symptoms, defined according to DSM-III-R criteria. OUTCOME MEASURES: A semistructured psychiatric interview using the Psychopathology Assessment Scale (AMDP-4) of the Association for Methodology and Documentation in Psychiatry system. Comprehensive medical assessments, including laboratory tests and computed tomographic scans, were also performed. RESULTS: Patients with cerebral opportunistic infections or metabolic encephalopathy (i.e., "secondary" psychosis, n = 13) were more likely to show disorders of consciousness, disorders of orientation and disturbances of attention and memory than those with no evidence of HIV-related cerebral disease (i.e., "primary" psychosis, n = 13); 10 patients (77%) with cerebral opportunistic infections or metabolic encephalopathy and only 1 (8%) patient without (p < 0.001) were diagnosed with delirium. These associations were stronger for the "secondary" patients with no focal brain lesions than for those with lesions. CONCLUSIONS: These findings suggest that "organic" symptoms of psychosis in those infected with HIV are related to the systemic and cerebral complications of HIV infection rather than to the psychotic process itself. PMID- 11394193 TI - Novelty-elicited mismatch negativity in patients with schizophrenia on admission and discharge. AB - OBJECTIVE: Given recent reports of differences between mismatch negativity (MMN) elicited by always novel sounds (novelty-elicited MMN) and that elicited by repeated rare deviants (conventional MMN), we investigated novelty-elicited MMN and P3a in patients with schizophrenia before and after a nonstandardized inpatient treatment. DESIGN: Electrophysiological and clinical assessment of patients on admission and discharge from hospital. Assessment of control subjects on 2 sessions. SETTING: Inpatient treatment in a psychiatric university hospital. SUBJECTS: 20 patients with schizophrenia and 21 healthy control subjects of similar age and sex. Selection of patients with first- to third-episode schizophrenia. OUTCOME MEASURES: Early and late component MMN amplitudes and latencies, P3a amplitudes and latencies, Positive and Negative Syndrome Scale (PANSS), Global Assessment of Functioning (GAF), Extrapyramidal Symptom Scale (EPS), Abnormal Involuntary Movement Scale (AIMS) and chlorpromazine equivalents. RESULTS: In patients with schizophrenia, novelty-elicited MMN was unimpaired on admission, and there was a statistically significant reduction of the late MMN component with treatment. Improvements in symptom expression were associated with increased latencies of the early MMN component. CONCLUSION: Results indicate differences in information processing between conventional and novelty-elicited MMN. Some components of the novelty-elicited MMN might be more state dependent than those of the conventional MMN. PMID- 11394195 TI - Bipolar disorder after mefloquine treatment. PMID- 11394194 TI - Response to pentagastrin after acute phenylalanine and tyrosine depletion in healthy men: a pilot study. AB - OBJECTIVE: To assess the effects of the acute depletion of the catecholamine precursors phenylalanine and tyrosine on mood and pentagastrin-induced anxiety. DESIGN: Randomized, double-blind controlled multiple crossover study. SETTING: University department of psychiatry. PARTICIPANTS: 6 healthy male volunteers. INTERVENTIONS: 3 treatments were compared: pretreatment with a nutritionally balanced amino acid mixture, followed 5 hours later by a bolus injection of normal saline placebo; pretreatment with a balanced amino acid mixture, followed by a bolus injection of pentagastrin (0.6 microgram/kg); and pretreatment with an amino acid mixture without the catecholamine precursors phenylalanine or tyrosine, followed by pentagastrin (0.6 microgram/kg). OUTCOME MEASURES: Scores on the panic symptom scale, a visual analogue scale for anxiety, the Borg scale of respiratory exertion and the Profile of Mood States Elation-Depression Scale. RESULTS: Pentagastrin produced the expected increases in anxiety symptoms, but there was no significant or discernible influence of acute phenylalanine and tyrosine depletion on anxiety or mood. CONCLUSIONS: These pilot data do not support further study using the same design in healthy men. Under these study conditions, phenylalanine and tyrosine depletion may have larger effects on dopamine than noradrenaline. Alternative protocols to assess the role of catecholamines in mood and anxiety are proposed. PMID- 11394196 TI - Clozapine and sialorrhea: update. PMID- 11394197 TI - A short form of the Wisconsin Card Sorting Test. PMID- 11394198 TI - Neural actions of luteinizing hormone and human chorionic gonadotropin. AB - Luteinizing hormone (LH) and its homologue, human chorionic gonadotropin (hCG), are able to elicit multiple effects in the central nervous system (CNS) through binding to their receptors. Specific receptors for LH/hCG have been identified in the hippocampus, dentate gyrus, hypothalamus, cortex, brain stem, area postrema, cerebellum, choroid plexus, ependymal cells, glial cells, neural retina, pituitary gland, and neuron processes of the spinal cord. Neurotropic effects of LH and hCG have been demonstrated in fetal rat brain, where the expression of LH/hCG receptors is developmentally regulated. Administration of hCG has been found to be beneficial in restoration of transected spinal cord function in rats. In adult rat brain, LH and hCG are involved in the feedback regulation of synthesis and secretion of gonadotropin-releasing hormone (GnRH) in the hypothalamus and LH in the pituitary gland. LH and hCG also induce several behavioral and other changes that are associated with the hippocampus, which contains the highest density of LH/hCG receptors. Many of the behavioral changes induced by hCG in rats parallel those in pregnant women. Some of these behavioral effects are correlated with changes of eicosanoid metabolism induced by LH and hCG in the brain. The LH/hCG receptors present in the choroid plexus, brain vessels, and perihypophyseal vascular complex may be involved in the modulation of transport of LH, hCG, and GnRH into the CNS. Thus, the CNS is one of the specific target tissues for LH and hCG, by which LH/hCG act as pleiotropic hormones that regulate several reproduction-related as well as reproduction nonrelated functions in the CNS. PMID- 11394199 TI - Rat as model for studying behavior effects of hCG. AB - The demonstration of receptors for luteinizing hormone (LH)/human chorionic gonadotropin (hCG) in several parts of rat brain suggested their novel functional role. Subsequent studies tested the effect of hCG (intraperitoneally or intracerebroventricularly) on brain arousal and different types of stress situations in an intact female rat model on the day of proestrus. Treatment resulted in changes of activity and several other behavioral patterns associated with sites of brain hCG/LH receptors. hCG given peripherally caused a longer sleeping time and a decreased activity level. Whereas administration of indomethacin alone had no effect, coadministration inhibited the effects of hCG. hCG increased immunoreactive prostaglandin D2 (PGD2) and decreased PGE2 in brain areas controlling activity and sleep. hCG effects were probably mediated via prostaglandin pathways. After central hCG treatment, animals were less active and showed less exploratory behavior in an open-field box than the control animals. Taste and odor neophobias were dramatically decreased following central injection of hCG. hCG-treated rats were less anxious and exhibited a higher level of maternal interest than the controls. hCG treatment also had a beneficial effect against stress ulcer, which was prevented by pretreatment with antisense receptor oligonucleotide, suggesting a direct hCG receptor-mediated effect. In summary, because hCG can cross the blood-brain barrier, both peripheral administration and central administration affect several behavioral patterns. This effect is similar to treatment with anxiolytics and suggests the functional relevance of brain LH/hCG receptors. Some observed behavioral changes have parallels in pregnant women. PMID- 11394200 TI - Nongonadal LH/hCG receptors in pig: functional importance and parallels to human. AB - Luteinizing hormone (LH) and human chorionic gonadotropin (hCG) share a common receptor in gonadal cells; however, the presence of these receptors has also been detected in several nongonadal but reproduction-associated tissues of pig, human, and other species. There are no data about the ontogeny of the human LH/hCG receptor. The expression of the porcine LH receptor gene in the uterus starts about 10 days after the appearance of this gene in gonads. LH/hCG receptors were found in uterus (myometrium, endometrium), oviduct, cervix, fetal membranes, and umbilical cord in humans and pigs. The main role of LH/hCG receptors in myometrium is stimulation of growth and hyperplasia and relaxation of uterine motility. hCG also increases blood flow in the uterine artery. LH and hCG can increase production of prostaglandins in endometrium, oviduct, and blood vessels. It is suggested that the preovulatory surge of LH plays an important role in controlling oviductal contractions. Human and pig mammary glands also possess LH/hCG receptors through which gonadotropins can affect the metabolism of steroid hormones in this tissue and may play an inhibitory role in mammary carcinogenesis and in the growth of breast tumors. PMID- 11394201 TI - Human chorionic gonadotropin: does it affect human endometrial morphology in vivo? AB - Human chorionic gonadotropin (hCG) is widely used as a surrogate to luteinizing hormone (LH) to trigger ovulation in controlled ovarian hyperstimulation (COH). Yet this molecule may exert direct effects on the endometrium. These effects, not mediated by ovarian hormones, are probably a consequence of stimulation of endometrial hCG/LH receptors. Because the half-life of hCG differs markedly from that of LH, possible pharmacological effects of hCG on the endometrium could alter endometrial receptivity in COH. Arguments supporting a clinical action of gonadotropins, and hCG in particular, on the endometrium abound. Notably, evidence has been reported of decidualization of stromal cells of the human endometrium in vitro as a result of exposure to gonadotropins, including hCG. The present article discusses, from a clinical standpoint, the main basis supporting the hypothesis that hCG administration as commonly used in infertility treatments may exert direct effects in vivo on endometrial histology and partake in endometrial transformations of the luteal phase. Preliminary results suggest that endometrial effects of hCG exist in vivo and should be taken into account when assessing the endometrial effects of hormones. PMID- 11394202 TI - On the role of human chorionic gonadotropin (hCG) in the embryo-endometrial microenvironment: implications for differentiation and implantation. AB - Recent evidence suggests that human chorionic gonadotropin (hCG), in addition to its well-known endocrine effects on the corpus luteum, may act as a growth and differentiation factor during pregnancy. According to experimental results, its mode of action may be divided into three sequential phases. During the first phase, which begins at the blastocyst stage and lasts until the occurrence in the serum, hCG acts preferentially in a juxtacrine manner. We have used an intrauterine microdialysis system developed in our laboratory to administer low concentrations of hCG to the endometrium of women in the luteal phase of the menstrual cycle. HCG administration provoked profound effects on paracrine parameters of differentiation (IGFBP-1, prolactin) and implantation (LIF, M-CSF). VEGF, a cytokine important for neoangiogenesis, was significantly stimulated by hCG (P < .01), suggesting a role for hCG in the control of endometrial vascularization and placentation. The investigation of endometrial parameters of tissue remodeling revealed a significant increase of MMP-9 (P < .05) but not of TIMP-1 following hCG infusion. The second, endocrine, phase of hCG action is marked by the appearance of hCG in the maternal serum. Rising systemic hCG levels cause a very rapid elevation of serum progesterone reflecting the rescue of the corpus luteum. Other endocrine functions of hCG include its intrinsic thyrotropic activity as well as modulation of fetal testicular, ovarian, and adrenal function. The third phase may be characterized by the expression of full-length hCG/LH receptors on the trophoblasts themselves. Before the ninth week of gestation, human villous trophoblasts express a truncated hCG/LH receptor isoform (50 kDa) and are probably not responsive to hCG. Later, the expression pattern is switched to the full-length receptor (80 kDa), allowing hCG also to modulate the differentiation of the trophoblasts themselves. A special feature is the self regulation of hCG biosynthesis that may in part explain the unique secretion profile of the hormone with peak levels during the first trimester followed by a rapid decline after the tenth week of gestation. In summary, hCG seems to have a variety of local and systemic functions in and outside the embryo-endometrial microenvironment. PMID- 11394203 TI - Effects of human chorionic gonadotropin on trophoblast invasion. AB - Recent studies have shown that different populations of trophoblastic cells of first-trimester human placenta express human chorionic gonadotropin/luteinizing hormone (hCG/LH) receptors. Among these trophoblastic cells, extravillous cytotrophoblasts (CTBs) not only express hCG/LH receptors but also present an invasive phenotype. CTB invasion is instrumental in blastocyst implantation and later in placentation and is tightly regulated in both time and space. This article describes some of the regulators of CTB invasion and summarizes and discusses the potential autocrine/paracrine role of hCG in trophoblast invasion. PMID- 11394205 TI - A proposed role for hCG in clinical obstetrics. AB - The role of human chorionic gonadatropin (hCG) in the maintenance of early pregnancy is well known. Recent data suggests that hCG may play a role in the maintenance of the later stages of pregnancy as well, by directly and indirectly promoting uterine quiescence. If hCG acts as an endogenous tocolytic in normal pregnancy, then it may be an ideal candidate for therapy of preterm labor as well. We present compelling in vitro as well as in vivo data, which support the role of hCG in the maintenance of normal uterine quiescence. Additionally, we will present in vivo and in vitro data that confirms the ability of hCG to directly promote relaxation of uterine contractions. This review provides a basis for future study of the use of hCG in clinical obstetrics. Given the limited effectiveness of tocolytic therapies available at the time, hCG may provide a promising pharmacological approach to the pervasive problem of preterm labor in human pregnancy. While further work is needed, initial data strongly support this novel use of hCG in clinical obstetrics. PMID- 11394204 TI - Clinical data supporting the importance of vascular LH/hCG receptors of uterine blood vessels. AB - Recent studies revealed functional extragonadal gonadotropin receptors at several sites of the human body. The human chorionic gonadotropin/luteinizing hormone (hCG/LH) receptor messenger RNA and protein were found in the endothelial and vascular smooth muscle layers of the human uterine arteries. In vivo administration of hCG decreased the blood flow resistance in the human uterus and in vitro increased vasodilating eicosanoids in the vascular wall. These findings initiated a further study investigating whether the administration of hCG to patients with signs of threatened abortion has any beneficial effect. The patients were treated with either magnesium or progesterone and/or hCG. The results showed that the rate of patients reaching second trimester was higher when hCG was included in the treatment protocol, and a parallel significant decrease in uterine blood flow resistance was also found. Analyzing the long-term results, the rate of preterm and growth-retarded deliveries was lower when hCG was administered in the first trimester. As a conclusion, the uterine vascular LH/hCG receptors play a significant role in the peri-implantation period. The hCG might also participate in angiogenesis, enhancing long-term clinical results. PMID- 11394206 TI - Uterine receptivity in the baboon: modulation by chorionic gonadotropin. AB - Embryonic signals not only rescue the corpus luteum but also modulate the uterine environment in preparation for implantation. Chorionic gonadotropin (CG), an early embryonic signal, is one such molecule. In vivo studies in the baboon show that CG alters the morphology and biochemical activity of the uterus, especially with respect to the three major cell types: luminal epithelium, glandular epithelium, and stromal fibroblasts. Further, CG and progesterone have a synergistic effect on the receptive endometrium. CG action on the endometrial cells is transduced via a seven transmembrane G protein-coupled receptor. Possible signaling pathways involved are discussed. PMID- 11394207 TI - An overview of the past, present, and future of nongonadal LH/hCG actions in reproductive biology and medicine. AB - Even though there were hints, it was not until 1986 that a number of laboratories worldwide began to demonstrate unequivocally the presence and functions of luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptors in various female and male nongonadal tissues. There was no species specificity but there was tissue specificity in the nongonadal LH/hCG receptor distribution. Nongonadal receptor levels were lower but they were regulated and processed and used signaling mechanisms similarly to gonadal receptors. Although still greater understanding is needed, gains made in the last decade demonstrate that nongonadal actions of LH/hCG are physiologically important and may have relevance to better understanding several diseases and their treatment. A recently developed LH receptor knockout model has begun to reaffirm and extend the importance of nongonadal LH signaling in the body. PMID- 11394208 TI - Use of the rat model to study hCG/LH effects on uterine blood flow. AB - Luteinizing hormone (LH) and human chorionic gonadotropin (hCG) share a common receptor. LH/hCG receptors, located in the vascular smooth muscle and endothelial cells of uterine blood vessels, are most numerous in smaller intramyometrial vessels and are cyclic in nature. There is a correlation between hCG levels and decreased uterine vascular resistance in humans, and in pseudopregnant rats, hCG decreases uterine blood flow. We found that systemic administration of hCG to cycling rats reduced uterine blood flow within 20 minutes on all days of the estrous cycle when flow was measured via the radioactive microsphere method. This effect was absent in ovariectomized rats. To determine the response to hCG at the microvascular level, we measured uterine arteriolar diameters in vivo via videomicroscopy after direct application or injection of hCG in rats on diestrus 1, diestrus-2, and proestrus. When hCG was suffused over the uterus (20 IU/60 mL), the uterine arterioles in diestrus (1 and 2) rats dilated but in proestrus rats were constricted. Neither response was altered in animals whose ovaries were removed 3 hours previously. An intraperitoneal injection of hCG (50 IU) caused uterine arteriolar constriction in diestrus-2 and proestrus animals but no change in diestrus-1. In ovariectomized rats, uterine arteriolar constriction following hCG injection was absent. Thus, the response of the uterine vasculature is not only cycle day dependent but also dependent on the route of administration. The effect of hCG on uterine blood flow in the rat may be a direct action of hCG, or it could be secondary to the release of other vasoactive substances known to be released by hCG. hCG might also alter blood flow by inducing angiogenesis. PMID- 11394209 TI - Functional importance of bovine myometrial and vascular LH receptors and cervical FSH receptors. AB - Bovine myometrium and cervix contain luteinizing hormone/human chorionic gonadotropin (LH/hCG) binding sites, LH receptor (LH-R) messenger RNA (mRNA), and LH-R protein. Expression of LH-R is dependent on the stage of the cycle. LH-R expression is high during the luteal phase but weak during the follicular phase. In both myometrium and cervix, LH activates both the adenylate cyclase and phospholipase C pathways, and the effect of LH on both pathways at each stage of the cycle is correlated with the amount of LH-R present in the tissue. Because activation of cyclic AMP (cAMP) is associated with myometrial quiescence, we suggest that LH activation of uterine cAMP could serve to keep the uterus quiescent during the luteal phase. On the other hand, in the uterine vein LH-R mRNA and LH-R are maximal during preestrus/estrus as opposed to the luteal phase. In the uterine vein, LH increases the expression of cyclooxygenase and production of both prostaglandin E2 (PGE2) and PGF2 alpha. Because PGF2 alpha is the physiological luteolytic signal in the cow, we suggest that this increase in prostaglandin production by the uterine vein is part of the physiological events leading to luteolysis. In addition to uterine LH-R, the bovine cervix at preestrus/estrus has high levels of follicle-stimulating hormone receptor (FSH-R) and its corresponding mRNA. As with LH-R, activation of FSH-R by FSH is associated with activation of a G protein-coupled receptor family that mediates the cAMP and inositol phosphate signaling pathways. Activation of these signaling pathways is associated with an increase in the expression of cyclooxygenase and production of PGE2. Because expression of the FSH receptor was maximal at the time of the FSH peak in the blood, we suggest a physiological role for FSH in the cervix relaxation and opening at estrus. PMID- 11394210 TI - A new function for the LH/CG receptor: transcytosis of hormone across the endothelial barrier in target organs. AB - Gonadotropins cross the vascular endothelial barrier to reach their target organs. Hormone concentrations in blood are very low and fluctuate rapidly to give sharp biological effects. Nonspecific transendothelial transport mechanisms are slow and markedly concentration dependent. There is thus a need for specific transendothelial transport mechanisms for these hormones. We will describe a specific transcytosis mechanism involving endothelial lutropin/chorionic gonadotropin (LH/CG) receptors identical to those observed in the target Leydig cells. A new role for the LH/CG receptor as a transcytotic carrier of its ligands in endothelial cells is suggested. PMID- 11394211 TI - Natural history and incidence of premature thelarche in Puerto Rican girls aged 6 months to 8 years diagnosed between 1990 and 1995. AB - OBJECTIVE: To describe the natural history and estimate the incidence of premature thelarche in girls aged 6 months to 8 years living in Puerto Rico and diagnosed between 1990 and 1995. BACKGROUND: In the 1970s and 1980s, several pediatric endocrinologists, based on their clinical experience, acknowledged a dramatic increase in the number of cases of precocious sexual development in Puerto Rico. In 1987, the Puerto Rico Health Department developed the Registry of Premature Thelarche and Precocious Sexual Development, which began to operate in 1989. Data regarding the long-term outcomes of girls diagnosed with premature thelarche are insufficient. Knowledge about the natural history of this condition is relevant for predicting the long-term prognosis and therapeutic management of the affected population. METHODS: Of 2,716 cases of precocious sexual development reported to the Premature Thelarche and Precocious Sexual Development Registry, 1,916 (70.5%) were premature thelarche. The clinical characteristics and evolution during follow-up of premature thelarche cases were described and compared by age group at diagnosis. RESULTS: Incidences were 6.2 and 1.62 per 1,000 live births for girls aged < 2 years and 2 to 8 years, respectively. These estimates were 10 and 15 times higher than those reported in Olmsted, MN. When the average change in mammary tissue diameter during follow-up was evaluated, a slight reduction in girls aged < 2 years was observed; however, it remained constant for girls aged 2 to 8 years. CONCLUSIONS: The results of this study underscore the need to continue an active search of premature thelarche cases and to perform analytical investigations of precocious sexual development to expand the understanding of the etiology of this important public health problem. PMID- 11394212 TI - Trends in diabetes mellitus mortality in Puerto Rico: 1980-1997. AB - OBJECTIVE: To determine the characteristics and trends of diabetes mortality among the Puerto Rican population from 1980 through 1997. METHODS: Death certificates for Puerto Rican residents whose underlying cause of death was diabetes mellitus (ICD-9-250.0) were reviewed, and sociodemographic information was abstracted. The proportion mortality ratio (PMR) and 95% confidence intervals were calculated by gender, age group, educational level and period of time. Trend analysis in mortality was performed using a Poisson regression model. RESULTS: A total of 26,193 deaths (5.8%) were primarily attributed to diabetes mellitus in the study period. Females accounted for 55.8% of all diabetes related deaths. Diabetes accounted for a higher proportion of deaths among persons aged 60-64 years (8.14%), persons aged 65-74 (8.12%), females (7.73%) and those with 1-6 years of education (7.08%). The PMR steadily increased from 4.55% in the 1980-85 period to 6.91% in the 1992-97 period. There was a higher mortality in male diabetic subjects aged < or = 64 than in females during the 18 year period. Between 1980 and 1991, females aged 65-74 had a higher mortality than males, however, mortality increased in males of the same age group during 1992-97. When the oldest age group (> or = 75) was examined, males had a higher mortality between 1986 and 1997, whereas females had a slightly higher rate between 1980 and 1985. CONCLUSIONS: Our results indicate that diabetes mortality has been markedly increasing in the Puerto Rican population, primarily in persons aged 65 years or more. Further analysis is needed to evaluate the determinants of mortality in diabetes. PMID- 11394213 TI - Preliminary evaluation of glipizide spheres and compacts from spheres prepared by cross-linking technique. AB - The objective of this research was to use the natural polymer Carrageenan to obtain controlled release spheres loaded with glipizide using the cross-linking technique. The effect of polymer level and drug load were investigated. The drug was dispersed in Carrageenan solution and the dispersion was dropped by a device containing 3 disposable syringes into cross-linking solution containing 3% calcium chloride. After 15 minutes residence time, the spheres were collected by decantation and dried in hot air oven at 38 degrees C +/- 2 degrees C for 24 hours. The dried spheres were successfully compacted into tablets using rotary Manesty B-3B machine equipped with 12/32 inches round flat face punches, target tablet weight was 400 mg +/- 5%. As the polymer level was increased in the sphere formulation, the drug release rate was increased. However, as the drug level was increased in the sphere formulation, the release rate was decreased. This trend was also true for tablets compacted from spheres. The scanning electron microscope photographs supported the dissolution data. More cracks and rough surface were observed in tablets compacted from spheres containing high polymer level and low drug level. PMID- 11394214 TI - Protein binding of glipizide using equilibrium dialysis technique: effects of hydrogen ion concentration, drug concentration and ionic strength. AB - The objective of this research was to investigate the effects of hydrogen ion concentration, drug concentration and ionic strength on the binding affinity of glipizide to albumin protein. Different buffer solutions of different pH values (pH 6.7, 7.5 and 8.5), different drug concentrations (2.45 mg, 4.82 mg and 9.42 mg), and phosphate buffer solutions pH 7.5 of different ionic strength (0.1, 0.4 and 1.0) were prepared. The effects of pH, drug concentration and ionic strength on the amount of glipizide bounded to 0.25 g bovine albumin was investigated. As the pH of the solution was increased from pH 6.4 to pH 8.5, milligrams drug bounded to gram protein (r value) decreased from 8.2 mg to 3.84 mg/g protein. Also as the ionic strength of the solution was increased from 0.1 to 1.0, the r value decreased from 10.76 mg to 3.96 mg/g protein. However, the r value did not change significantly with increasing of drug from 2.45 mg to 9.42 mg/25 ml. The r value was 7.36 mg/g protein when concentration of the drug was 2.45 mg/25 ml and 7.4 mg/g protein when the concentration of the drug was 9.42 mg/25 ml. This study demonstrated that factors such as high pH and high ionic strength can alter drug protein binding and consequently increase free drug in plasma and increase bioavailability of slightly water insoluble drug such as antidiabetic drugs. PMID- 11394215 TI - Screening, monitoring, and educating patients with diabetes in an independent community pharmacy in Puerto Rico. AB - OBJECTIVE: Increase the awareness about the importance of Diabetes mellitus (DM) management and assess the educational and monitoring needs of patients visiting a community pharmacy in Puerto Rico. METHODS: A community service activity focusing on DM was held in a community pharmacy. The educational and monitoring needs of the participants were assessed using a questionnaire. Glucose tests were conducted in the pharmacy by medical technologists. Educational activities consisted of presentations and printed materials. RESULTS: Two-thirds of the fasting people had blood glucose levels higher than 140 mg/dl. Seventy-nine percent of the patients with diabetes were not aware of the glycosilated hemoglobin test. Most of the patients were interested in learning more about how to manage their condition. CONCLUSION: A greater understanding is needed among patients with DM that blood glucose control decreases diabetes related complications. Community pharmacists are in an excellent position to collaborate with other health professionals in screening, monitoring and educating patients with DM to prevent long-term complications. PMID- 11394216 TI - [Analysis on the availability of programs and services for elderly people in Puerto Rico]. AB - This research was conducted with the purpose of analyzing the programs and services available to the aged population in the municipalities of Puerto Rico during 1999 and 2000. The analysis was performed using an evaluation instrument which collected data of the programs and services directed exclusively to the aged population, such as: senior centers, foster homes, institutions, independent living, employed and/or volunteer people, geriatric services, adult education (literacy), recreation, discounts, corporations and/or associations that benefit the aged population, aging offices, support groups, social services, home meal services, and financial aid. Data was also collected from programs and services that benefit diverse aged populations in areas such as: health, housing, employment and social welfare. The sources for the collection of data were directories available to government and private agencies, telephone directories, telephone interviews, and personal visits. The municipalities were divided into two zones, the Northern zone, constituted by 40 municipalities and the Southern zone, constituted by 38 municipalities, for a total of 78 evaluated municipalities. In the Northern region, which accounted to approximately 295,938 aged persons, 18 types of programs or services were found to serve the aged population directly and 50 types were aged-related programs or services. In the Southern region, which accounted for approximately 169,798 aged persons, 13 types of programs or services served the aged exclusively and 43 were aged-related services. The following were among the principal programs or services available: senior centers, foster homes and/or institutions, employment programs, home care services and hospices, independent living, and agricultural extension services. Among the least available programs or services were private home meal services, literacy and retraining, arts and travel services, corporations or associations for the benefit of the aged, geriatric evaluation programs, home repair services, home-assistance programs, support and respite programs, adequate or specialized transportation services, diverse housing models, different types of senior centers, and programs for psychological or psychiatric assistance. In conclusion, the availability of the programs and services in this study and the evaluation of the services needed for a community of aged persons indicate the need for increasing and developing services considered essential in accordance to the population demands. PMID- 11394217 TI - Predictors of adverse events after percutaneous transluminal coronary angioplasty in a group of Hispanic patients. AB - OBJECTIVE: To identify predictors of adverse events after PTCA during hospitalization and after hospital discharge in a private hospital in Puerto Rico. BACKGROUND: A review of the literature shows limited information about predictors of adverse events associated to percutaneous transluminal coronary angioplasty (PTCA) in Hispanic patients. METHODS: This is a non-concurrent prospective study. Baseline variables were analyzed using multivariate logistic regression to identify predictors of adverse events. Data were collected from medical charts and telephone reports from referring physicians. RESULTS: Data from 197 subjects undergoing PTCA were analyzed for this study. Median age of patients was 65 years, and 62.9% of patients were male. Angiographic success rate was 81.6%. A total of 8.1% of patients had at least one in-hospital adverse event, and 39.8% had at least one adverse event after hospital discharge. After multivariate analysis, a statistically significant association was found between the presence of at least one lesion with residual stenosis of 50% or greater and the risk of developing adverse events in-hospital (RO 11.75; 95% CI 4.32-31.97). A marginally significant association was found between family history of heart disease (RO 2.75; 95% CI 0.93-8.11) and the risk of adverse events during hospitalization. Family history of heart disease (RO 1.41; 95% CI 0.98-2.04) and the presence of at least one lesion with residual stenosis of 50% or greater (RO 2.87; 95% CI 0.82-10.01) showed marginally significant associations with increased risk for adverse events after discharge. CONCLUSIONS: These findings suggest that the presence of at least one lesion with residual stenosis of 50% or greater and family history of heart disease may be risk factors for adverse events after PTCA during hospitalization and after discharge. PMID- 11394218 TI - Youth violence: understanding and prevention: strategies of intervention. Part II. AB - Youth violence is a complex problem. The recent youth related violent incidents are of great impact taking in consideration the emotional costs to victims, their families and to the health and safety of citizens; as well as the economic cost to society. It is a major public health concern that requires participation, collaboration and integration of efforts from parents, citizens and professionals from different disciplines. PMID- 11394219 TI - Puerto Rican health teachers: attitudes towards breastfeeding. AB - OBJECTIVE: The purpose of the study was to identify attitudes towards breastfeeding and support for breastfeeding in public in a group of health teachers in the Department of Education. METHOD: The study design was correlational descriptive. A self-administered questionnaire was used (Cronbach's alpha = 0.83) for 125 health teachers. Descriptive and inferential statistics (chi square and t test) were used for data analysis. RESULTS: 89.6% were women, 47.1% had 39 years of age or less, 76.4% were married, the median of years in the profession was 12.5. Only 8.8% had breastfed exclusively, 46.1% used artificial feedings exclusively, and 45.1% combined artificial milk and breast milk. A moderate/negative attitude towards breastfeeding was shown by 53.1%. 60.3% stated they agreed or totally agreed that in order to breastfeed the mother must follow a specific diet, 36.0% agreed or totally agreed that breast milk should alternate with artificial milk, and 100% of participants do not support breastfeeding in public. No significant difference was found in the attitude scale towards breastfeeding and the gender, the age, years in the profession, and the type of milk given their children. CONCLUSIONS: We must train teachers in the Department of Education, on a priority basis, in the field of human lactation in view of their importance for health promotion. PMID- 11394220 TI - [Reflections on the human genome]. PMID- 11394221 TI - [Testimonials of brave women: the forgotten voices]. AB - This paper examines the narrative testimony of women survivors of two atrocious events that took place in the XX century: the Holocaust in the 40's and the HIV/AIDS pandemic in the 80's. The author recognizes similarities in thoughts, feelings, experiences and meaning regarding several issues of human suffering that emerge from these testimonies. It expounds the perceptions on death, motherhood, family separation, intimacy and sexuality. For the author approach of these issues from a female perspective can provide new meanings arrived at the development of a new discourse and new social practices. Promotes to think about reactions of indifference before human suffering. Concludes by questioning why technological, scientific advances and advances in social development have not been able to provide human responses to human problems. PMID- 11394222 TI - [Historical notes on the foundation of the medical school of the University of Puerto Rico]. PMID- 11394223 TI - [Historical recollection Medical School of the University of Puerto Rico 1950 2000]. AB - The School of Medicine of the University of Puerto Rico was founded in 1950 with the mission of educating the much needed physician workforce in order to improve the health of a large portion of the population in Puerto Rico. The main events in its first fifty years of existence are summarized. Emphasis is given to the unique, mutually dependent association between the School of Medicine and the Department of Health of Puerto Rico. Soon after its organization, the school became a principal protagonist in the delivery of specialized medical care to the medically indigent population within the existing Regionalization Program of Health Care services in the island. With the creation and development of various other academic and health services institutions in the island, and the advent of a new system of health care in 1993, based on managed care; the School's interdependence with the Department of Health and its role in the direct care of the medically indigent have waned drastically. The School now faces its greatest challenges as it begins to insert itself into the economically competitive arena of the new health care system; and in redefining its commitments, while searching for new resources, alliances, teaching faculty, hospitals and clinics, enabling it to maintain its leadership in medical education, specialty training and scientific research in Puerto Rico. PMID- 11394224 TI - [A historical sketch of the Medical School of the University of Puerto Rico]. PMID- 11394225 TI - [Occurrence of congenital defects in the Czech Republic from 1996 to 1999 in the offspring of mothers taking medications during the 1st trimester of pregnancy]. AB - OBJECTIVE OF STUDY: Elaborate the results of recorded inborn defects in mothers taking drugs during the first trimester of pregnancy. TYPE OF STUDY: Retrospective analysis of data from register of inborn defects in the Czech Republic. METHOD: Elaboration of the incidence of selected types of inborn defects assessed in children born to mothers taking drugs during the first trimester of pregnancy. Data from the nationwide register of inborn defects kept in the Institute for Health Information and Statistics of the Czech Republic in 1996-1999 were used. From this total number 519 cases were recorded in born infants and 31 in foetuses. In born infants there were 215 affected girls and 304 affected boys. Some of the inborn defects were recorded more frequently in our material than in the remaining population. RESULTS: In 1996-1999 in our retrospective analysis a total of 550 cases of inborn defects were recorded in women taking medication during the first trimester of pregnancy. CONCLUSION: The paper presents recent results of the incidence of nationwide registration of inborn defects during a four-year period. PMID- 11394226 TI - [Analysis of stillbirths in the Czech Republic in 1998]. AB - OBJECTIVE: To identify the reasons for stagnation of stillbirth rate in the Czech Republic and to evaluate possibilities how to avoid some cases of stillbirth. SETTING: Institute for the Care of Mother and Child, Prague 4. DESIGN: Retrospective analysis of individual cases of stillbirth. METHODS: Retrospective analysis of all cases of stillbirth without congenital malformations in the Czech Republic during 1998 was performed by regional perinatologists according to unified guidelines. The analysis targeted the evaluation of perinatal management with respect to possible flaws in care and with respect to avoidability of stillbirth. We prepared a questionnaire to collect data with special attention to birthweight and type of the facility (level of care) where death occurred. The data collected were analyzed in the Institute for the Care of Mother and Child. RESULTS: Out of 252 cases of stillbirth (contribution to total perinatal mortality of 2.8 per 1000) only 0.2 per 1000 were among multiple pregnancies; 0.13 per 1000 were deaths during labor. We found mistakes in the content of care among 40% of cases, problems in organization of care were in every second case of stillbirth, and combination of both was frequent. Every sixth case of death had insufficient prenatal care due to pregnant women. Probable avoidability was higher among foetuses with birthweight of 2000 grams and higher. Cases of mistakes in care and avoidability was higher in terrain (out of hospital) in absolute terms, however, related to number of cared population, the mistakes and avoidability were higher in hospitals. This is related to higher concentration of complicated pregnancies in these facilities. CONCLUSIONS: Based on the final evaluation of possible avoidability of some cases of stillbirth we can conclude that even in the case of ultimate success of prenatal care the contribution of stillbirth to decrease of total perinatal mortality would not exceed 0.6 per 1000; the cost of such care would increase substantially. PMID- 11394227 TI - [Perinatal outcome of twin pregnancies after fertilization in vitro and after spontaneous conception]. AB - OBJECTIVE: To compare the obstetric outcome of twin pregnancies after IVF with spontaneously conceived twins and thus determine whether IVF twins require greater care. TYPE OF STUDY: A retrospective study. SETTING: The Department of Obstetrics and Gynaecology, 1st Faculty of Medicine of Charles University, Prague. METHODS: Statistical evaluation of obstetric outcome between the group of 46 twin pregnancies after IVF and 85 spontaneously conceived twins. We evaluated the following parameters: signs of abortion and premature delivery, bleeding in pregnancy, premature rupture of membranes, performed cerclage, incidence of praeclampsia, gestational diabetes mellitus and hepatopathy of the mother. Pathological ultrasound findings were also evaluated--disturbances in foetal growth and the amount of amniotic fluid. In these parameters we also evaluated the time of the first signs of their development. Further, we evaluated the type of delivery, the gestational age at delivery, birth weight, birth weight discordance and perinatal mortality. RESULTS: We found statistically significant differences in the method of delivery--more cesarean sections in the IVF group (71.7%) than in spontaneous ones (44.7%). Another difference was the gestational age at which cerclage were performed--earlier in the IVF group (average 22.8 weeks) than in spontaneous ones (average 25.7 weeks). In the other evaluated parameters we did not find any statistically significant differences between the two groups. CONCLUSION: In our retrospective analysis of twin pregnancies, deliveries and neonatal outcome, except for the method of delivery and time of performing cerclage, we did not find any important significant differences between the IVF and spontaneously conceived twins. However, it is necessary to remember the generally worse obstetric outcome of twin compared to singleton pregnancies. PMID- 11394228 TI - [Interleukin-6, procalcitonin, C-reactive protein and the immature to total neutrophil ratio (I/T) in the diagnosis of early-onset sepsis in low birth weight neonates]. AB - OBJECTIVE: To determine the influence of early-onset neonatal sepsis on interleukin-6 (IL-6) and procalcitonin (PCT) levels in cord blood. To evaluate the significance of usually used infection markers--C-reactive protein (CRP) and immature to total neutrophil ratio (I/T)--and new markers (PCT, IL-6) for the diagnosis of early-onset neonatal sepsis. DESIGN: Prospective clinical study. SETTING: Institute for the Care of Mother and Child, Prague. METHODS: The serum levels of IL-6 and PCT were measured in cord blood in 37 low birht weight infants less than 35 week of gestation born in our institute. IL-6 and PCT levels were further evaluated together with CRP and I/T in neonatal blood within 2 hours after delivery. Neonatal sepsis within the first 72 hours of life was monitored. RESULTS: Differences in mean values of CRP, I/T, IL-6, and PCT between "sepsis proven" and "sepsis not proven" groups were not statistically significant. Only the difference between groups in cord blood PCT was of borderline significance (p = 0.06, higher in "sepsis proven" group). Fisher test showed significant dependence on sepsis in cord blood PCT only (cut-off point 0.4 ng/ml, p < or = 0.05). Other parameters did not show significant dependence on sepsis. Sensitivity for early onset sepsis above 50% was found in cord blood PCT only (sensitivity 60%, specificity 85.2%). PCT predictive accuracy for sepsis expressed as AUC value was 0.74 +/- 0.06. CONCLUSION: The only relatively sensitive marker and moderate predictor of early-onset sepsis in premature low birthweight infant was in our study cord blood PCT. PMID- 11394229 TI - [Embryo-fetoscopy--present possibilities of endoscopy in obstetrics]. AB - OBJECTIVE: The introduction of the embryofetoscopy--a new method of prenatal diagnosis used in the 1st trimester of the pregnancy. SETTING: Institute for the Care of Mother and Child, Prague. DESIGN: Pilot clinical study. METHODS: The embryofetoscopy was carried out in pregnant women in the 7th or 8th conceptional week who asked for a legal abortion of the pregnancy. We used the embryofetoscope with diameter of 0.8 mm (made by K. Storz, Tuttlingen, Germany). The trocar of the embryofetoscope was inserted into the animal cavity transabdominally always guided by ultrasound. It is possible to perform the examination in local anaesthesia. A further trocar with two canals--one for insertion of the embryofetoscope, the second of the needle with diameter of 0.6 mm--is designated for genetic indicated sampling of embryonal tissues or for incidental application of gene or stem cells therapy in the future. RESULTS: The videocamera documented our embryoscopical findings till this time in 7 cases. On the pictures 1-8 are the face, scull, eye, ear, extremities, genitals and umbilical cord of the embryo in the 7th conceptional week. CONCLUSIONS: The applications of the embryofetoscopy are very wide. It allows us to obtain not only new knowledge about the early development of the normal and affected embryo in vivo, but it simultaneously opens new ways for the therapy of the embryo in the future and for fetal surgery guided by embryoscope in the 2nd and 3rd trimester of the pregnancy. PMID- 11394230 TI - [Validity of cardiotocography in acute fetal hypoxia--neonatal status after cesarean section]. AB - OBJECTIVE: Judging the validity of CTG monitoring as a method of acute foetal hypoxia diagnosis (AFH), quantifying the number of cesarean section (SC), performed from the indication of acute fetal hypoxia and point out a number of surgeries based on false positive CTG results. DESIGN: Retrospective study. SETTING: 1st Department of Obsterics and Gynaecology Faculty of Medicine MU, Brno. METHODS: 100 women, patients of the 1st Department of Obstetrics and Gynaecology in Obilni trh, Brno, who had cesarean section due to foetal hypoxia indicated on the base of pathological or suspect CTG, were involved in the collection. In the next stage newborns were divided into 2 clinical groups: "Acidosis" and "Normal" depending on the pH value from a. umbilicalis and statistically analysed. RESULTS: From 100% suspect or pathological CTG, only 36.19% were valid (newborns showed depression after birth). Remaining 63.18% of newborns, although showed CTG signs of hypoxia were born normal and probably were not necessary to be born by cesarean section. CONCLUSION: Although CTG investigation is in most places the only "objective" method for diagnosis of AFH, this investigation cannot precisely determinate whether hypoxia is present or it is not. For true objectivity of foetal intrauterine condition and correct indication of SC is suitable to use further diagnostic methods of AFH in future, for example intrapartal foetal pulse oximetry or blood analysis from foetal scalp. PMID- 11394231 TI - [Effect of natural surfactant on mortality and morbidity in neonates with a birth weight below 1500 grams]. AB - OBJECTIVE OF STUDY: Evaluation of the natural surfactant in treatment of the respiratory distress syndrome (RDS) on the mortality and morbidity of neonates with a birth weight lower than 1500 g. TYPE OF STUDY: Retrospective analysis of data. SETTING: Department:perinatological centre, Institute for Mother and Child, Prague. METHOD: A total of 442 neonates with birth weights under 1500 g were evaluated. During period I/1994-XII/1998 271 neonates were analyzed (Group A). The results of the total neonatal mortality and morbidity were compared with results of care provided to 171 patients during I/1989-XII/1993 before introduction of the surfactant. Surfactant therapy was used in 134 patients of group A (9%) with RDS who met the standard indication criteria. The initial surfactant dose (Alveofact R Boehringer Ingelheim, GFR) depended on the ventilation parameters and was 50 mg/kg and in severe forms of RDS 100 mg/kg. The total cumulative dose did not exceed 200 mg/kg. The data were assembled in a database used in the Institute for the Care of Mother and Child and evaluated using the statistical programme EpiInfo (CDC, USA). RESULTS: In the group of neonates with a birth weight lower than 1000 g 110 neonates in group A were evaluated and 61 in group B. Surfactant was administered to 77 infants of the treated group (70%). Statistically significant differences were recorded: in the total mortality (A: 29 vs. B: 79%, p < 0.0001, the number of neonates who died during UPV (A: 32% vs. B: 80%, p < 0.0001), incidence of PVH-IVH grade I-IV (A: 29% vs. B: 67%, p < 0.0001). In the group of neonates with a birth weight of 1000 to 1499 g 161 children were evaluated in group A and 110 in group B. Alveofact was administered to 57 neonates of the treated group (35%). Statistically significant differences were recorded: in the total mortality (A: 4%, vs. B: 31%, p < 0.0001), the number of neonates who died during UBPV (A: 7% vs. B: 45%, p < 0.0001), incidence of perintraventricular grade I-IV (A: 11% vs. B: 35%, p < 0.0001), incidence of patent ductus arteriosus (A: 6% vs. B: 15%, p < 0.02), incidence of pneumothorax (A: 7% vs. B: 18%, p < 0.01) and in the incidence of posthaemorrhagic hydrocephalus (A: 2% vs. B: 11%, p < 0.01). There was a statistically significant difference in the incidence of BPD to the disadvantage of group A (A: 16% vs. B: 8%, p < 0.01). CONCLUSION: The study confirmed the favourable effect of the introduction of surfactant in the treatment of RDS and on the general mortality and morbidity of neonates with a birth weight below 1500 g. PMID- 11394232 TI - [Neonatal mortality in the Czech Republic 1998-1999]. AB - OBJECTIVE: To evaluate neonatal mortality rate (NMR) in 1998 and 1999 years in the Czech Republic. DESIGN: Retrospective epidemiological study of all alive newborns born in 1998 and 1999 in the Czech Republic. SETTINGS: 12 perinatological centers of nine regions of Bohemia and Moravia. METHODS: All alive, died, died with congenital defects newborns were registered and results of neonatal mortality rate and specific neonatal mortality rate were calculated. The main causes of death were divided into four groups (intraventricular hemorrhage grade III-IV, infection, acute respiratory failure and others) and evaluated comparatively. In 1999 the NMR of newborns with birth weight below 500 g and their survival were introduced for the first time in the Czech Republic. RESULTS: The fluent decrease of NMR during nineties was stopped in 1999. Increase of NMR from 2.8@1000 in 1998 to 3.0@1000 in 1999 was mainly caused by arise of specific neonatal mortality rate in newborns weighing > or = 2000 g. Comparing 1998 and 1999 years, two times more these newborns without serious congenital defects died in 1999 (28 vs. 56). Specific neonatal mortality rate of extremely low birth weight newborns further decreased (359@1000 vs. 279@1000) especially in the newborns with birth weight between 500-749 g (543@1000 vs. 373@1000). The most frequent main causes of death still has been intraventicular haemorrhage grade III-IV and infection in very low birth weight newborns, and serious congenital defects and infection in newborns weighing > or = 1500 g. The concentration of very low birth weight newborns to perinatological centers by transfer in uterus was 81% in 1998 and 83% in 1999. The differences in neonatal mortality rates between nine regions of Bohemia and Moravia has been getting equal but has been still great in specific neonatal mortality rate of extremely low birth weight newborns between the best and worst regions (147@1000 vs. 458@1000). There were registered 19 newborns weighing < or = 500 g surviving more than 24 hours after delivery in the Czech republic. Specific neonatal mortality rate of these newborns was 316@1000 and 527@1000 survived. CONCLUSION: Reserves for further lowering of NMR are improving the care after extremely low birth weight newborns in the regions with below average results and decreasing the mortality of newborns with birth weight > or = 2000 g by introducing of group B streptococcus prophylaxis, improving prenatal diagnostics of serious congenital defects and early and more quality postnatal transport of newborns suffered from acute respiratory failure to centers disposing of the latest methods of treatment. PMID- 11394233 TI - [Extracorporeal membrane oxygenation in the treatment of acute respiratory failure in full-term neonates]. AB - OBJECTIVE OF STUDY: Evaluation of success of extracorporeal membrane oxygenation, EMCO) in the treatment of acute respiratory failure in mature neonates. TYPE OF STUDY: Clinical pilot study of the EMCO centre. SETTING: Institute for the Care of Mother and Child, Prague. MATERIAL AND METHOD: In 12 consecutive patients with severe acute respiratory distress syndrome and/or circulatory failure of different etiology who met the indication criteria the method of venoarterial EMCO was used. RESULTS: Venoarterial extracorporeal membrane oxygenation was successful in 75% patients who survived. Four patients died (1x syndrome of cerebral death associated with severe hypoxic-ischaemic encephalopathy, 2x severe irreversible haemorrhage, into the abdominal and thoracic cavity, 1x periventricular intraventricular haemorrhage grade III.). The mean period of EMCO was 71.4 +/- 31.7 hours (range 25-130 hours). On comparison of the surviving group (S) and the non-surviving group (NS) there was a significant difference in the necessity of continuous inotropic adrenaline support during EMCO. In patients who died necropsy confirmed irreversible multiorgan failure. CONCLUSION: In indicated cases extracorporeal membrane oxygenation remains the method of choice in critically ill mature neonates. A prognostically adverse factor is the necessity of inotropic support and haemodialysis during EMCO. PMID- 11394234 TI - [Problems in inducing pulmonary maturation in relation to premature delivery]. AB - At presents efforts are made to find valid early markers of premature delivery and if it is inevitable to use an optimal therapeutic protocol for induction of pulmonary maturity which may lead to reduction of early neonatal mortality and morbidity (intraventricular-periventricular haemorrhage, necrotizing enterocolitis, bronchopulmonary dysplasia, air-leak syndrome, respiratory distress syndrome--RDS). Hitherto used corticoid monotherapy reduced the incidence of RDS in neonates with a birth weight lower than 1500 g but the incidence is still relatively high and RDS is still one of the main causes of death of immature neonates. Even the use of thyrotropic hormones did not reduce its incidence. Another possible therapeutic approach is ambroxol administration in combination with corticoids. In the presented review the authors discuss the problem of optimal induction of pulmonary maturity in relation to premature delivery. PMID- 11394235 TI - [Occurrence of cleft defects of the central nervous system in the Czech Republic 1961-1999. Incidence, prenatal diagnosis and prevalence based on maternal age]. AB - OBJECTIVE OF STUDY: Presentation of the incidence of neural tube defects (anencephaly, spina bifida, encephalocele) in the Czech Republic in 1961 to 1999. Analysis of the prevalence of these defects in different groups by maternal age. TYPE OF STUDY: Retrospective demographic-epidemiological study. METHOD: Epidemiological analysis of the incidence of neural tube defects diagnosed prenatally and postnatally in the Czech Republic in 1961-1999. Mathematical and statistical analysis of these defects by maternal age, the method of 95% confidence probability interval was used. Data from the nationwide register of inborn defects were used assembled in the Institute of Health Information and Statistics of the Czech Republic and data on the prenatal diagnosis from different departments of medical genetics. RESULTS: In the Czech Republic in 1961 1999 4629 cases of neural tube defects were recorded. This number comprised 1812 cases of anencephaly, 2420 cases of spina bifida and 397 cases of encephalocele. Of this total number of notified defects 386 cases of anencephaly were diagnosed prenatally and pregnancy was terminated. In spina bifida thus 261 pregnancies were terminated prematurely, in encephalocele 67 cases. CONCLUSION: A significant reduction of the incidence of anencephaly, spina bifida and encephalocele in the Czech Republic was found during the last decade of the investigation period (1961 1999). This decline is most probably due to advances of primary and secondary prevention. When evaluating maternal age as one of the risk factors for the development of neural tube defects, this risk was neither unequivocally confirmed nor ruled out. PMID- 11394236 TI - Strenuous endurance training in humans reduces oxidative stress following exhausting exercise. AB - The aim of this study was to evaluate whether high-intensity endurance training would alleviate exercise-induced oxidative stress. Nine untrained male subjects (aged 19-21 years) participated in a 12-week training programme, and performed an acute period of exhausting exercise on a cycle ergometer before and after training. The training programme consisted of running at 80% maximal exercise heart rate for 60 min.day-1, 5 days.week-1 for 12 weeks. Blood samples were collected at rest and immediately after exhausting exercise for measurements of indices of oxidative stress, and antioxidant enzyme activities [superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT)] in the erythrocytes. Maximal oxygen uptake (VO2max) increased significantly (P < 0.001) after training, indicating an improvement in aerobic capacity. A period of exhausting exercise caused an increase (P < 0.01) in the ability to produce neutrophil superoxide anion (O2.-) both before and after endurance training, but the magnitude of the increase was smaller after training (P < 0.05). There was a significant increase in lipid peroxidation in the erythrocyte membrane, but not in oxidative protein, after exhausting exercise, however training attenuated this effect. At rest, SOD and GPX activities were increased after training. However, there was no evidence that exhausting exercise enhanced the levels of any antioxidant enzyme activity. The CAT activity was unchanged either by training or by exhausting exercise. These results indicate that high-intensity endurance training can elevate antioxidant enzyme activities in erythrocytes, and decrease neutrophil O2.- production in response to exhausting exercise. Furthermore, this up-regulation in antioxidant defences was accompanied by a reduction in exercise induced lipid peroxidation in erythrocyte membrane. PMID- 11394237 TI - Measurement and prediction of peak shivering intensity in humans. AB - Prediction equations of shivering metabolism are critical to the development of models of thermoregulation during cold exposure. Although the intensity of maximal shivering has not yet been predicted, a peak shivering metabolic rate (Shivpeak) of five times the resting metabolic rate has been reported. A group of 15 subjects (including 4 women) [mean age 24.7 (SD 6) years, mean body mass 72.1 (SD 12) kg, mean height 1.76 (SD 0.1) m, mean body fat 22.3 (SD 7)% and mean maximal oxygen uptake (VO2max) 53.2 (SD 9) ml O2.kg-1.min-1] participated in the present study to measure and predict Shivpeak. The subjects were initially immersed in water at 8 degrees C for up to 70 min. Water temperature was then gradually increased at 0.8 degree C.min-1 to a value of 20 degrees C, which it was expected would increase shivering heat production based on the knowledge that peripheral cold receptors fire maximally at approximately this temperature. This, in combination with the relatively low core temperature at the time this water temperature was reached, was hypothesized would stimulate Shivpeak. Prior to warming the water from 8 to 20 degrees C, the oxygen consumption was 15.1 (SD 5.5) ml.kg-1.min-1 at core temperatures of approximately 35 degrees C. After the water temperature had risen to 20 degrees C, the observed Shivpeak was 22.1 (SD 4.2) ml O2.kg-1.min-1 at core and mean skin temperatures of 35.2 (SD 0.9) and 22.1 (SD 2.2) degrees C, respectively. The Shivpeak corresponded to 4.9 (SD 0.8) times the resting metabolism and 41.7 (SD 5.1)% of VO2max. The best fit equation predicting Shivpeak was Shivpeak (ml O2.kg-1.min-1) = 30.5 + 0.348 x VO2max (ml O2.kg-1.min-1) - 0.909 x body mass index (kg.m-2) - 0.233 x age (years); (P = 0.0001; r2 = 0.872). PMID- 11394238 TI - Blood lactate response to overtraining in male endurance athletes. AB - Many physiological markers vary similarly during training and overtraining. This is the case for the blood lactate concentration ([La-]b), since a right shift of the lactate curve is to be expected in both conditions. We examined the possibility of separating the changes in training from those of overtraining by dividing [La-]b by the rating of perceived exertion ([La-]b/RPE) or by converting [La-]b into a percentage of the peak blood lactate concentration ([La-]b,peak). Ten experienced endurance athletes increased their usual amount of training by 100% within 4 weeks. An incremental test and a time trial were performed before (baseline) and after this period of overtraining, and after 2 weeks of recovery (REC). The [La-]b and RPE were measured during the recovery of each stage of the incremental test. We diagnosed overtraining in seven athletes, using both physiological and psychological criteria. We found a decrease in mean [La-]b,peak from baseline to REC [9.64 (SD 1.17), 8.16 (SD 1.31) and 7.69 (SD 1.84) mmol.l-1, for the three tests, respectively; P < 0.05] and a right shift of the lactate curve. Above 90% of maximal aerobic speed (MAS) there was a decrease of mean [La ]b/RPE from baseline to REC [at 100% of MAS of 105.41 (SD 17.48), 84.61 (SD 12.56) and 81.03 (SD 22.64) arbitrary units, in the three tests, respectively; P < 0.05), but no difference in RPE, its variability accounting for less than 25% of the variability of [La-]b/RPE (r = 0.49). Consequently, [La-]b/RPE provides little additional information compared to [La-]b alone. Expressing [La-]b as a %[La-]b,peak resulted in a suppression of the right shift of the lactate curve, suggesting it was primarily the consequence of a decreased production of lactate by the muscle. Since the right shift of the curve induced by optimal training is a result of improved lactate utilization, the main difference between the two conditions is the decrease of [La-]b,peak during overtraining. We propose retaining it as a marker of overtraining for long duration events, and repeating its measurement after a sufficient period of rest to make the distinction with overreaching. PMID- 11394239 TI - Evidence for neuromuscular fatigue during high-intensity cycling in warm, humid conditions. AB - The purpose of this study was to examine and describe the neuromuscular changes associated with fatigue using a self-paced cycling protocol of 60-min duration, under warm, humid conditions. Eleven subjects [mean (SE) age 21.8 (0.8) years; height 174.9 (3.0) cm; body mass 74.8 (2.7) kg; maximum oxygen consumption 50.3 (1.8) ml.kg.min-1] performed one 60-min self-paced cycling time trial punctuated with six 1-min "all out" sprints at 10-min intervals, while 4 subjects repeated the trial for the purpose of determining reproducibility. Power output, integrated electromyographic signal (IEMG), and mean percentile frequency shifts (MPFS) were recorded at the mid-point of each sprint. There were no differences between trials for EMG variables, distance cycled, mean heart rate, and subjective rating of perceived exertion for the subjects who repeated the trial (n = 4). The results from the repeated trials suggest that neuromuscular responses to self-paced cycling are reproducible between trials. The mean heart rate for the 11 subjects was 163.6 (0.71) beats.min-1. Values for power output and IEMG expressed as a percentage of that recorded for the initial sprint decreased during sprints 2-5, with normalised values being 94%, 91%, 87% and 87%, respectively, and 71%, 71%, 73%, and 77%, respectively. However, during the final sprint normalised power output and IEMG increased to 94% and 90% of initial values, respectively. MPFS displayed an increase with time; however, this was not significant (P = 0.06). The main finding of this investigation is the ability of subjects to return power output to near initial values during the final of six maximal effort sprints that were included as part of a self-paced cycling protocol. This appears to be due to a combination of changes in neuromuscular recruitment, central or peripheral control systems, or the EMG signal itself. Further investigations in which changes in multiple physiological systems are assessed systematically are required so that the underlying mechanisms related to the development of fatigue during normal dynamic movements such as cycling can be more clearly delineated. PMID- 11394240 TI - Force recovery after eccentric exercise in males and females. AB - In this study we investigated force loss and recovery after eccentric exercise, and further characterized profound losses in muscle function (n = 192 subjects- 98 males, 94 females; population A). Maximal voluntary contractile force (MVC) was assessed before, immediately after, and at 36 and 132 h after eccentric exercise. Two groups were then established (A1 and A2). Group A1 demonstrated a > 70% reduction in MVC immediately after exercise, but were recovering at 132 h after exercise. These subjects performed a follow-up MVC 26 days later (n = 32). Group A2 demonstrated a > 70% reduction in MVC immediately post-exercise, but still exhibited a > 65% reduction in force at 132 h post-exercise; these subjects also performed a follow-up MVC every 7 days until full recovery was established (n = 9). In population A, there was a 57% reduction in MVC immediately post exercise and a 67% recovery by 132 h post-exercise (P < 0.01), with no significant gender differences (P > 0.05). In group A1, although more females (two-thirds) showed large force losses after exercise, these females demonstrated greater %MVC recovery at 132 h post-exercise (59% vs 44%) and at 26 days post exercise (93% vs 81%) compared to the males. In group A2, MVC recovery occurred between 33 and 47 days post-exercise. In conclusion, 21% of all subjects showed a delayed recovery in MVC after high-force eccentric exercise. Although there were no significant gender differences in force loss, a disproportionately larger number of females demonstrated force reductions of > 70%. However, their recovery of force was more rapid than that observed for the males who also demonstrated a > 70% force loss. PMID- 11394241 TI - Changes in myosin heavy chain composition with heavy resistance training in 60- to 75-year-old men and women. AB - The purpose of this investigation was to assess the myosin heavy chain (MHC) expression in the vastus lateralis muscle from elderly men and women, and to determine whether heavy resistance training influences its expression. Twenty healthy, mildly physically active subjects gave their informed consent to participate in the study. The experimental group consisted of seven men and seven women [mean (SD) age 65.5 (4.1) years] and the control group consisted of three men and three women [mean (SD) age 62.3 (3.6) years]. The 6-month resistance training program was divided into two phases with weeks 1-12 consisting of high intensity resistance training, and weeks 13-24 involving power training. Muscle biopsy samples were taken from the vastus lateralis muscle at week 0 and week 24 using the needle biopsy technique. The male and female experimental groups both exhibited a significant decrease (P < or = 0.05) in the percentage of MHC IIb, while the experimental female group also demonstrated a significant increase (P < or = 0.05) in the expression of MHC IIa, after 24 weeks of heavy resistance training. There was no change in MHC expression within the control group. The male [130.4 (25.3) kg vs 171.1 (30.5) kg] and female [58.2 (8.3) kg vs 77.9 (11.1) kg] experimental groups exhibited a significant increase (P < or = 0.05) in the maximal strength values for the 1 repetition maximum (1RM) squat exercise. The control group showed no change in strength for the 1RM squat exercise for either the male [115.8 (35.10 kg vs 123.8 (47.2) kg] or female [57.5 (99.0) kg vs 58.3 (2.9) kg] groups. The results clearly show that elderly subjects undergoing heavy resistance training have the ability to produce a similar shift in the expression of MHC isoforms from MHC IIb to MHC IIa, as has been shown to occur in younger subjects. This highlights the plasticity of human skeletal muscle in response to heavy resistance training, even at older ages. PMID- 11394242 TI - The effect of heavy resistance exercise on the circadian rhythm of salivary testosterone in men. AB - Circadian rhythms of serum testosterone concentrations in men have been shown, in general, to be highest in the morning and lowest in the evening. Thus, the purpose of this investigation was to determine the effects of acute resistance exercise upon the waking circadian rhythm of salivary testosterone over 2 days (with or without resistance exercise). The subjects included ten resistance trained men (with at least 1 year of lifting experience) with the following characteristics [mean (SD)]: age 21.6 (1.1) years; height 177.8 (9.5) cm; body mass 80.5 (11.5) kg; percent body fat 7.9 (1.7)%. A matched, randomized, crossover study design was used such that each subject was tested under both the resistance exercise and control (no exercise) conditions. The resistance exercise protocol consisted of ten exercises performed for three sets of ten repetitions maximum with 2 min of rest between sets. Saliva sample 1 was collected at 0615 hours and resistance exercise began immediately afterwards at approximately 0620 hours, and sample 2 was collected at 0700 hours, which corresponded approximately to a mid-exercise (or control) time point. Saliva samples were then obtained every hour on the hour from 0800 hours until 2200 hours. No significant differences were observed between the exercise and resting conditions for salivary testosterone, with the exception of a significant decrease at 0700 hours during the resistance exercise protocol. The results of this investigation indicate that resistance exercise does not affect the circadian pattern of salivary testosterone secretion over a 16-h waking period in resistance-trained men. PMID- 11394243 TI - The human force:velocity relationship; activity in the knee flexor and extensor muscles before and after eccentric practice. AB - The human voluntary force:velocity relationship frequently fails to demonstrate the expected high eccentric forces. Possible explanations include unique activation strategies which might be affected by neural learning mechanisms. We investigated the effect of practicing eccentric contractions on (1) the force:velocity relationship of the human knee extensor muscles and (2) the extent of agonist and antagonist muscle activity. Eight healthy adults [seven women, group mean age 31 (SEM 5) years +/-] practiced twice a week for 4 weeks using their non-dominant legs. Each session comprised three isokinetic concentric and eccentric maximal voluntary contractions (MVC) at randomised angular velocities of 100, 200 and 300 degrees.s-1. Before and after, the force:velocity relationship was determined bilaterally (angular velocities 0-300 degrees.s-1). There were no significant differences in the forces generated or relative electromyogram (EMG) activity after practice, although there was a trend for dynamic forces to increase. Beforehand, the bilateral eccentric MVC forces were lower than isometric (P < 0.0025); afterwards they were broadly similar. The agonist EMG was similar during isometric and eccentric contractions, but lower during concentric (P < 0.03). Antagonist EMG activity showed considerable individual variation, was similar during all contraction types and tended to be greater during dynamic contractions. These data indicate that neither central learning mechanisms nor total muscle activation strategies underlie the human failure to produce the expected high eccentric voluntary forces in humans. PMID- 11394244 TI - Exercise-induced oxidative stress and muscle performance in healthy women: role of vitamin E supplementation and endogenous oestradiol. AB - The purpose of this study was to investigate the individual and combined antioxidant effects of menstrual cycle phase-related alterations in blood serum oestradiol concentrations and of dietary vitamin E supplementation on exercise induced oxidative stress and muscle performance. A group of 18 sedentary women, aged 19-35 years, were given supplements of 300 mg alpha-tocopherol (n = 10) or placebo (n = 8) daily during the course of two menstrual cycles. The subjects exercised the knee isokinetically to exhaustion after cycling submaximally at 50% maximal oxygen uptake during the menstrual and preovulatory phases of their menstrual cycles. Blood samples were taken before and after the exercise, to evaluate haematocrit, plasma lactic acid and malondialdehyde concentrations, erythrocyte antioxidant enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities and apolipoprotein B containing lipoprotein (non-high density lipoprotein, HDL, fraction) oxidation. Serum vitamin E, follicle stimulating hormone, luteinizing hormone and oestradiol concentrations were measured in pre-exercise blood samples. Neither vitamin E supplementation nor oestradiol concentrations influenced SOD and GPx activities or the susceptibility of the non-HDL fraction to oxidation while at rest. Plasma malondialdehyde concentration was unaffected by exercise, however significant reductions in erythrocyte SOD and GPx activities and increased susceptibility of the non-HDL fraction to oxidation were noted after exercise. Exercise-induced changes were reduced when oestradiol concentration was high in the preovulatory phase, independent of the serum vitamin E concentrations. In addition, both pre- (r = 0.58, P < 0.05) and post-exercise (r = 0.73, P < 0.001) GPx activities in placebo administered subjects were positively correlated with oestradiol concentrations. In conclusion, these findings suggest a better protective role of oestradiol against oxidative injury, compared to vitamin E. Exhausting muscle performance was, however, not influenced by vitamin E supplementation and/or cycle-phase related changes in oestradiol concentrations. PMID- 11394245 TI - Effect of strenuous strength training on the Na-K pump concentration in skeletal muscle of well-trained men. AB - This study examined how strenuous strength training affected the Na-K pump concentration in the knee extensor muscle of well-trained men and whether leg muscle strength and endurance was related to the pump concentration. First, the pump concentration, taken as 3H-ouabain binding, was measured in top alpine skiers since strength training is important to them. Second, well-trained subjects carried out strenuous eccentric resistance training either 1, 2, or 3 times.week-1 for 3 months. The Na-K pump concentration, the maximal muscle strength in a full squat lift (one repetition maximum, 1 RM), and the muscle endurance, taken as the number of full squat lifts of a mass of 70% of the 1 RM load, were measured before and after the training period. The mean pump concentration of the alpine skiers was 425 (SEM 11) nmol.kg-1 wet muscle mass. The subjects in part two increased their maximal strength in a dose-dependent manner. The muscle endurance increased for all subjects but independently of the training programme. From a mean starting value of 356 (SEM 6) nmol.kg-1 the mean Na-K pump concentration increased by 54 (SEM 15) nmol.kg-1 (+15%, P < 0.001) when the results for all subjects were pooled. The effect was larger for those who had trained twice a week than for those who had trained only once a week (P = 0.025), suggesting that the effect of strength training depended on the amount of training carried out. The muscle strength and endurance were not related to the pump concentration, suggesting that the pumping power of this enzyme did not limit the performance during heavy lifting. However, the individual improvements in the endurance test during the training period correlated with the individual changes in the pump concentration (rSpearman = 0.5; P = 0.01) which could mean that a common factor both increases the pump concentration and makes the muscles more adapted to repeated heavy lifting. PMID- 11394246 TI - Lack of IL-6 production during exercise in patients with mitochondrial myopathy. AB - In the present study we investigated the possibility that exercise-induced increases in plasma levels of interleukin (IL)-6 are associated with plasma lactate levels. Patients with mitochondrial myopathy (MM) are characterised by high levels of plasma lactate. In this study, seven patients with MM underwent an ergometer cycle test for 25 min without treatment. They were then treated with dichloroacetate (DCA) for 15 days. DCA inhibits pyruvate-dehydrogenase-kinase, thereby increasing the activity of the pyruvate-dehydrogenase complex. The same exercise test was repeated on the last day of treatment. DCA lowered the plasma lactate and increased plasma IL-6 concentrations at rest. IL-6 increased in response to exercise only during DCA treatment. Furthermore, plasma IL-6 was negatively correlated to plasma lactate at rest (r = -0.786, P = 0.05). Given that IL-6 is a cytokine with growth-promoting potential, the results of this study suggest that high lactate production contributes to the decreased muscle function observed in MM patients by inhibiting the production of IL-6. PMID- 11394247 TI - Do the kinetics of peripheral muscle oxygenation reflect systemic oxygen intake? AB - To examine whether the kinetics of local muscle oxygenation reflect systemic oxygen intake, we measured the kinetics of local muscle oxygenation and systemic oxygen consumption (VO2). This study included 16 healthy males who performed an exercise tolerance test on a bicycle ergometer. During the exercise test, expiratory gas analysis was performed with an expiratory gas analyzer, and the kinetics of vastus lateralis muscle oxygenation were determined by near-infrared spectroscopy (NIRS). Oxygenated hemoglobin (OxyHb) and tissue blood oxygen saturation (StO2) gradually decreased during the exercise test, while deoxygenated hemoglobin (DeoxyHb) gradually increased. We examined correlations between the mean values of these parameters, which were calculated by time integrating the values obtained using NIRS and dividing them by the integral time, and VO2. There was a marked positive correlation between DeoxyHb and VO2 (r = 0.893-0.986), and a marked negative correlation between StO2 and VO2 (r = 0.859 0.995). There was a negative correlation between VO2 and OxyHb (r = 0.726-0.978), and no correlation between TotalHb and VO2. These results suggest that the kinetics of peripheral muscle oxygenation reflect systemic VO2. PMID- 11394248 TI - Effects of repeated muscle contractions on the tendon structures in humans. AB - The purpose of this study was to investigate the changes in the elastic properties of tendons in humans in relation to fatigue of knee extensor muscles. The muscle fatigue test (MFT) consisted of maximal isometric contractions performed 50 times. The decline in peak moment was 43.6 (SD 19.5)%. After MFT, the muscle thickness and pennation angle of the vastus lateralis muscle (VL) significantly increased 1.5 (SD 0.7) mm (5%) and 1.7 (SD 1.8) degrees (11%), respectively. Before and after MFT, the elongation (l) of the tendon and aponeurosis of VL was directly measured by ultrasonography, while the subjects performed ramp isometric knee extensions up to maximal voluntary contraction. The l tended to be greater after MFT than before MFT. This difference in the l was statistically significant (P < 0.05) at force developments beyond 220 N. Furthermore, the compliance increased significantly from 2.0 (SD 0.6).10(-2) mm.N 1 before MFT to 2.6 (SD 0.7).10(-2) mm.N-1 after MFT (22.7%). In addition, the electromechanical delay was significantly increased from 60.6 (SD 5.9) ms before to 70.0 (SD 4.4) ms after MFT. These results suggested that the repeated muscle contractions made the tendon structures more compliant. PMID- 11394249 TI - Mechanomyogram and force relationship during voluntary isometric ramp contractions of the biceps brachii muscle. AB - The aim of the present study was to examine the non-stationary mechanomyogram (MMG) during voluntary isometric ramp contractions of the biceps brachii muscles using the short-time Fourier transform, and to obtain more detailed information on the motor unit (MU) activation strategy underlying in the continuous MMG/force relationship. The subjects were asked to exert ramp contractions from 5% to 80% of the maximal voluntary contraction (MVC) at a constant rate of 10% MVC/s. The root mean squared (RMS) amplitude of the MMG began to increase slowly at low levels of force, then there was a slight reduction between 12% and 20% MVC. After that, a progressive increase was followed by a decrease beyond 60% MVC. As to the mean power frequency (MPF), a relatively rapid increase up to 30% MVC was followed by a period of slow increment between 30% and 50% MVC. Then temporary reduction at around 50% MVC and a further rapid increase above 60% MVC was observed. The interaction between amplitude and MPF of the MMG in relation to the MU activation strategy is discussed for five force regions defined on the basis of their inflection points in the RMS-amplitude/force and MPF/force relationships. It was found that the MMG during ramp contractions enables deeper insights into the MU activation strategy than those determined during traditional separate contractions. In addition, this contraction protocol is useful not only to ensure higher force resolution in the MMG/force relationship, but also to markedly shorten the time taken for data acquisition and to reduce the risk of fatigue. PMID- 11394250 TI - Adaptation of the short latency component of the stretch reflex plays only a minor role in compensating for muscle fatigue induced by spontaneous hopping in humans. AB - The influence of fatigue on the stretch reflex evoked in ankle extensor muscles by hopping was investigated in six healthy men. The men hopped on a force platform, at spontaneous frequency and amplitude, until they were unable to maintain the initial frequency or amplitude of the jumps. This task was done with the knees flexing normally during ground contact or under instructions to straighten the knees. Surface electromyograms (EMG) of soleus (SO), gastrocnemius medialis (GM), and tibialis anterior (TA) muscles were recorded simultaneously with the vertical component of the ground reaction force. Spectrum analysis of the EMG recorded during isometric tests performed immediately before and after the fatiguing hopping task demonstrated the existence of fatigue in SO and GM and often in TA. The stretch reflex was studied during the first and last ten jumps of every hopping series. The long-latency components of the reflex were too variable to be analysed. Whatever the hopping condition, latency and amplitude of the short latency component were not significantly modified by fatigue. Fatigue enhanced the occurrence of this reflex component in SO only. These data suggest that in fatiguing submaximal hopping, the neuromuscular system does not fundamentally change its stiffness regulation before the endurance time has been reached. PMID- 11394251 TI - Effects of knee joint angles and fatigue on the neuromuscular control of vastus medialis oblique and vastus lateralis muscle in humans. AB - The effects of different knee joint angles and fatigue on the neuromuscular control of the vastus medialis oblique (VMO) and vastus lateralis (VL) muscles were investigated in 17 (11 men, 6 women) young subjects. The electromyogram (EMG) activities and the force generation capacities were monitored before and after a fatigue protocol at three different knee joint angles, 90 degrees, 150 degrees, 175 degrees of knee extension, on three occasions. In response to randomly triggered light signals, the subjects performed three isometric maximal voluntary contraction (IMVC) that lasted for 4 to 8 s. This was then followed by the fatigue protocol which consisted of six bursts of contractions fixed at 30 s on and 10 s off. Immediately after the exercise to fatigue, the subjects performed another three IMVC in response to the light signals. Repeated measures ANOVA were performed to examine the effects of fatigue at these three positions on the electromechanical delay (EMD), median frequency (fmed), peak force (Fpeak) and root mean square (rms)-EMG:Fpeak quotient of VMO and VL. The results revealed a significant effect of the three knee joint angles on the EMD before the fatigue (P < 0.05). The fatigue protocol induced a significant decrease in Fpeak at all the three positions (P < 0.01). However, the fatigue induced a significant decrease of fmed at only 90 degrees and 150 degrees of knee extension (P < 0.01). This occurred in parallel with the lengthening of EMD at these two joint angles (P < 0.01 and P < 0.05). The effects of fatigue on the fmed and EMD were not significant between VMO and VL at all three angles. The insignificant difference in fmed and EMD between VMO and VL at the three knee positions before and after fatigue indicated that no preferential onset activation between VMO and VL had occurred. PMID- 11394252 TI - Cortisol and testosterone concentrations in wheelchair athletes during submaximal wheelchair ergometry. AB - It is yet unknown how upper body exercise combined with high ambient temperatures affects plasma testosterone and cortisol concentrations and furthermore, how these hormones respond to exercise in people suffering spinal cord injuries. The purpose of this study was to characterize plasma testosterone and cortisol responses to upper body exercise in wheelchair athletes (WA) compared to able bodied individuals (AB) at two ambient temperatures. Four WA [mean age 36 (SEM 13) years, mean body mass 66.9 (SEM 11.8) kg, injury level T7-T11], matched with five AB [mean age 33.4 (SEM 8.9) years, mean body mass 72.5 (SEM 13.1) kg] exercised (cross-over design) for 20 min on a wheelchair ergometer (0.03 kg resistance.kg-1 body mass) at 25 degrees C and 32 degrees C. Blood samples were obtained before (PRE), at min 10 (MID), and min 20 (END) of exercise. No differences were found between results obtained at 25 degrees C and 32 degrees C for any physiological variable studied and therefore these data were combined. Pre-exercise testosterone concentration was lower (P < 0.05) in WA [18.3 (SEM 0.9) nmol.l-1] compared to AB [21.9 (SEM 3.6) nmol.l-1], and increased PRE to END only in WA. Cortisol concentrations were similar between groups before and during exercise, despite higher rectal temperatures in WA compared to AB, at MID [37.21 (SEM 0.14) and 37.02 (SEM 0.08) degrees C, respectively] and END [37.36 (SEM 0.16) and 37.19 (SEM 0.10) degrees C, respectively]. Plasma norepinephrine responses were similar between groups. In conclusion, there were no differences in plasma cortisol concentrations, which may have been due to the low relative exercise intensities employed. The greater exercise response in WA for plasma testosterone should be confirmed on a larger population. It could have been the result of the lower plasma testosterone concentrations at rest in our group. PMID- 11394253 TI - Are the blood pressure and endocrine responses of healthy subjects exposed to cold stress altered by an acutely increased sodium intake? AB - In the study reported here, we examined blood pressure and endocrine responses in cold conditions during salt load in young healthy subjects who had previously shown increased resting blood pressure during acutely increased sodium intake. Subjects (n = 53) added 121 mmol sodium into their normal diet for 1 week. If their mean arterial pressure had increased by a minimum of 5 mmHg compared to the previous measure they were selected for subsequent experiments. The subjects (n = 8) were given 121 mmol supplemental sodium.day-1 for 14 days. They were then put into a wind tunnel for 15 min (temperature--15 degrees C, wind speed 3.5.ms-1). Their blood pressure increased (P < 0.05) during the cold exposure, independent of the sodium intake. Their mean (SEM) plasma noradrenaline increased from 3.58 (0.62) nmol.l-1 to 5.61 (0.79) nmol.l-1 (P < 0.05) when the subjects were given a normal diet, and from 2.45 (0.57) nmol.l-1 to 5.06 (0.56) nmol.l-1 (P < 0.05) when the subjects were given an elevated sodium diet. The starting concentrations and the endpoint concentrations were statistically similar. The plasma levels of the N-terminal fragment of pro-atrial natriuretic peptide decreased during the whole-body cold exposure: with the sodium load the change was from 256.6 (25.5) nmol.l-1 to 208.0 (25.3) nmol.l-1, and with the normal diet, from 205.8 (16.4) nmol.l-1 to 175.1 (16.1) nmol.l-1. The haematocrit and red blood cell count increased (P < 0.05) with normal and elevated sodium diet in cold conditions, but haemoglobin increased (P < 0.05) only with high salt in cold conditions. To conclude, acutely increased sodium intake does not change the blood pressure response or hormonal responses to exposure to acute cold stress in healthy subjects. PMID- 11394254 TI - Changes in human skeletal muscle contractility and hormone status during 2 weeks of heavy strength training. AB - To examine neuromuscular and hormone changes during 2 weeks of heavy strength training, 18 weight-trained male students were recruited either into a heavy training group (HT, n = 11) or into a control group (Ctr, n = 7). The heavy training protocol consisted of leg-extensor workouts performed daily, while workouts were performed twice a week in the Ctr group. A test of one repetition maximum (1 RM) was performed before heavy training and on the 2nd day after heavy training. Isokinetic knee extensions, electrical stimulation, and squat jumps were performed before, on the 8th day of heavy training, and on the 4th day after heavy training. Morning blood samples (0800 hours) were drawn before, on the 8th day of heavy training, and on the 4th day after heavy training. Before, and on the 5th day after heavy training, 24 h urine samples were collected. The 1 RM leg press increased by 6 (SEM 2)% in the HT group. Testosterone and insulin-like growth factor-1 concentrations were respectively 12 (SEM 5)% and 11 (SEM 3)% lower than baseline on the 8th day of heavy training; however, hormone levels were back to baseline on the 4th day after heavy training. A significant correlation between individual changes in 1 RM leg press and changes in testosterone concentrations was observed in the HT group (r = 0.69). In the HT group, 24 h urinary catecholamine excretion increased by 26 (SEM 12)%, 3 methylhistidine excretion increased by 21 (SEM 6)% and creatinine excretion increased by 11 (SEM 5)%. There were no significant changes in the Ctr group. This work addresses the role of changes in basal hormone status (morning samples) for skeletal muscle adaptation to heavy strength training. PMID- 11394255 TI - Heart rate during aerobics classes in women with different previous experience of aerobics. AB - This study measured heart rate during floor and step aerobic classes at three intensity levels. A group of 20 female occasional exercisers [mean age 33 (SD 8) years, mean body mass index 21 (SD 2) kg.m-2 volunteered to participate in six aerobic classes (three floor classes, three step classes) and in a laboratory test as members of one of two groups according to their prestudy regular participation in aerobics classes. Subjects in group A had participated four or more times a week and those of group B less than twice a week. The characteristics of the groups were as follows: group A, n = 10, mean maximal oxygen uptake (VO2max) 38.7 (SD 3.6) ml.kg-1.min-1, mean maximal heart rate (HRmax) 183 (SD 8) beats.min-1; group B, n = 10, VO2max 36.1 (SD 3.6) ml.kg-1.min 1, HRmax 178 (SD 7) beats.min-1. Each class consisted of a warm-up, a 20 min period of structured aerobic exercise (cardiophase) and a cool-down. The cardiophase was planned and guided as light, (rate of perceived exertion, RPE 11 12), moderate (RPE 13-14) or heavy (RPE 15-17) by an experienced instructor. The mean heart rates during the light classes were 72 (step) and 74 (floor) %HRmax in group A and 75 (step) and 79 (floor) %HRmax in group B; during the moderate classes, 84 (step) and 80 (floor) %HRmax in group A and 82 (step) and 83 (floor) %HRmax in group B, and during the heavy classes 89 (step and floor) %HRmax in group A and 88 (step) and 92 (floor) %HRmax in group B. Differences in heart rate and %HRmax were not statistically significant between the groups. However, differences in heart rate and %HRmax between the intensities (light vs moderate, moderate vs heavy and light vs heavy) were significant within both groups (all, P < 0.01). Based on the results, we conclude that intensity management during the aerobics classes was generally successful regardless of the participants' prior participation in aerobics. However, some individuals who were older and/or had less prior participation tended to exceed the targeted heart rate. PMID- 11394256 TI - Heat of sorption induced by sweating affects thermoregulatory responses during heat load. AB - To investigate the effects of heat of sorption per se induced by sweating on thermoregulatory responses in clothed subjects, two experiments were carried out. In experiment 1 [ambient temperature (Ta) 27.2 degrees C, 50% relative humidity (r.h.)], seven female subjects immersed their lower-legs in a water bath at a temperature raised between 35-41 degrees C during 70 min wearing garments made for the experiment of either 100% cotton (C) or 100% polyester (P). Skin blood flow at the forearm was significantly greater in C than in P (P < 0.05). The increase of mean skin temperature (Tsk) and clothing surface and clothing microclimate temperatures were significantly higher in C than that of P after the onset of sweating (P < 0.05). Furthermore, these responses were accompanied by warmer and more uncomfortable sensations in C than in P despite a lower rectal temperature in C compared with P and identical mean body temperature in both sets of garments. In experiment 2, to simulate the clothing microclimate after the onset of sweating, C and P garments were exposed in the climate chamber to r.h. raised from 50% to 95% at a constant Ta of 27.2 degrees C. The clothing surface temperature rose by 2.2 degrees C for C and by 0.5 degree C for P after the increase of r.h. These results clearly showed a more marked increase in heat of sorption in C than in P. These results indicated that the heat of sorption per se after the onset of an increasing clothing microclimate vapour pressure, mimicking the onset of sweating, enhanced thermoregulatory responses such as skin blood flow, (Tsk) and subjective voting in C. PMID- 11394257 TI - Changes in muscle size, architecture, and neural activation after 20 days of bed rest with and without resistance exercise. AB - Nine healthy men carried out head-down bed rest (BR) for 20 days. five subjects (TR) performed isometric, bilateral leg extension exercise every day, while the other four (NT) did not. Before and after BR, maximal isometric knee extension force was measured. Neural activation was assessed using a supramaximal twitch interpolated over voluntary contraction. From a series cross-sectional magnetic resonance imaging scans of the thigh, physiological cross-sectional areas (PCSA) of the quadriceps muscles were estimated (uncorrected PCSA, volume/estimated fibre length). Decrease in mean muscle force after BR was greater in NT [-10.9 (SD 6.9)%, P < 0.05] than in TR [0.5 (SD 7.9)%, not significant]. Neural activation did not differ between the two groups before BR, but after BR NT showed smaller activation levels. Pennation angles of the vastus lateralis muscle, determined by ultrasonography, showed no significant changes in either group. The PCSA decreased in NT by -7.8 (SD 0.8)% (P < 0.05) while in TR PCSA showed only an insignificant tendency to decrease [-3.8 (SD 3.8)%]. Changes in force were related more to changes in neural activation levels than to those in PCSA. The results suggest that reduction of muscle strength by BR is affected by a decreased ability to activate motor units, and that the exercise used in the present experiment is effective as a countermeasure. PMID- 11394258 TI - Extracellular pH defense against lactic acid in normoxia and hypoxia before and after a Himalayan expedition. AB - The extracellular pH defense against the lactic acidosis resulting from exercise can be estimated from the ratios -delta[La].delta pH-1 (where delta[La] is change in lactic acid concentration and delta pH is change in pH) and delta[HCO3-].delta pH-1 (where delta[HCO3-] is change in bicarbonate concentration) in blood plasma. The difference between -delta[La].delta pH-1 and delta[HCO3-].delta pH-1 yields the capacity of available non-bicarbonate buffers (mainly hemoglobin). In turn, delta[HCO3-].delta pH-1 can be separated into a pure bicarbonate buffering (as calculated at constant carbon dioxide tension) and a hyperventilation effect. These quantities were measured in 12 mountaineers during incremental exercise tests before, and 7-8 days (group 1) or 11-12 days (group 2) after their return from a Himalayan expedition (2800-7600 m altitude) under conditions of normoxia and acute hypoxia. In normoxia -delta[La].delta pH-1 amounted to [mean (SEM)] 92 (6) mmol.l-1 before altitude, of which 19 (4), 48 (1) and 25 (3) mmol.l-1 were due to hyperventilation, bicarbonate and non-bicarbonate buffering, respectively. After altitude -delta[La].delta pH-1 was increased to 128 (12) mmol.l-1 (P < 0.01) in group 1 and decreased to 72 (5) mmol.l-1 in group 2 (P < 0.05), resulting mainly from apparent large changes of non-bicarbonate buffer capacity, which amounted to 49 (14) mmol.l-1 in group 1 and to 10 (2) mmol.l-1 in group 2. In acute hypoxia the apparent increase in non-bicarbonate buffers of group 1 was even larger [140 (18) mmol.l-1]. Since the hemoglobin mass was only modestly elevated after descent, other factors must play a role. It is proposed here that the transport of La- and H+ across cell membranes is differently influenced by high-altitude acclimatization. PMID- 11394259 TI - Combined effect of heat stress, dehydration and exercise on neuromuscular function in humans. AB - This study examined the combined effect of exercise induced hyperthermia and dehydration on neuromuscular function in human subjects. Six trained male runners ran for 40 min on a treadmill at 65% of their maximal aerobic velocity while wearing a tracksuit covered with an impermeable jacket and pants to impair the evaporation of sweat. These stressful experimental running conditions led the runners to a physiological status close to exhaustion. On average, the 40 min run ended at a heart rate of 196 (SD 8) beats.min-1, a tympanic temperature of 40 (SD 0.3) degrees C and with a loss of body mass of 2 (SD 0.5)%. Pre- and post-running strength tests included measurements of maximal knee extension and flexion torques in both isometric and isokinetic (at 60 and 240 degrees.s-1) conditions. A 20 s endurance test at 240 degrees.s-1 was also performed. Surface electromyographic (EMG) activity was recorded from six knee extensor and flexor muscles during the entire protocol. The treadmill run led to clear decrements in maximal extension torque and EMG activity both in isometric and at the slowest isokinetic velocity (60 degrees.s-1). However, no differences in these parameters were observed at 240 degrees.s-1. Furthermore, the EMG patterns of the major knee extensor and flexor muscles remained remarkably stable during the treadmill run. These results demonstrate that the exercise-induced hyperthermia and dehydration in the present experiments had only minor effects on the neuromuscular performance. However, it is also suggested that high internal body temperature per se could limit the production of high force levels. PMID- 11394260 TI - The VO2 slow component in swimming. AB - All studies on the oxygen uptake (VO2) slow component have been carried out for the sporting disciplines of cycling or running, but never for swimming. Considering that front crawl swimming is a sport discipline that is fundamentally different from both running and cycling, the aim of this study was to verify whether this slow component also appears in swimming. Six elite pentathletes were tested in a swimming flume while front crawl swimming to exhaustion. Swimming velocity for the slow component test was determined as v50% delta = CV + [vVO2peak - CV)/2], where CV is the critical velocity and vVO2peak the lowest velocity at which peak VO2 occurred. To set the subject's CV, expressed as the slope of a straight line that describes the correlation between swimming distance and time, the record times over three swimming distances were recorded in a 50 m swimming pool. The vVO2peak was measured by means of an incremental test in the swimming flume. Gas exchange was measured by means of a telemetric metabolimeter (K4 RQ, Cosmed, Italy) that was connected to a snorkel. The slow component was found in all subjects, with a mean (SD) value of 239 (194) mlO2.min-1. Therefore, although front crawl swimming is fundamentally different from both running and cycling, it appears that it also incurs a VO2 slow component. The origin of this phenomenon, however, is even more uncertain than for the other sport disciplines. PMID- 11394261 TI - Development of dissolved air flotation technology from the first generation to the newest (third) one (DAF in turbulent flow conditions). AB - This paper gives a brief description of the development of dissolved air flotation DAF (or so-called high pressure flotation) as an unit operation for removal of solids in water and wastewater treatment during the last 80 years up to this time. The first DAF-systems used in the water industry were the ADKA and Sveen-Pedersen ones from the 1920s. Some of these are still in use. The tanks in which the flotation phenomenon takes place in these systems are very shallow and narrow as well as rather long. The flow rate of water is some 2-3 m/h (at most less than 5 m/h only) and there is a very thin micro-bubble blanket below the water surface between the dry sludge blanket on that and the clarified water which flows almost horizontally below the bubble blanket toward the end of the tanks to be taken out there from near the bottom. The second generation of DAF was introduced in the 1960s and these units are widely in use today. Their tanks are almost square ones having usually a little bit more length than breadth. They are rather deep, too. There is an under-flow wall in front of the back wall of the units having a narrow horizontal gap on the bottom of the tanks for letting out the clarified water from the flotation space. The flow rate of water is usually 5-7 m/h or at most less than 10 m/h. The direction of flow is 30-45 degrees below the horizontal. There is a rather thick micro-bubble bed at the beginning of the tank below the dry sludge blanket. This bubble-bed becomes clearly thinner, when going toward the end of the tank. There are also round DAF tanks which are based on the same hydraulic principles as the rectangular ones presented above. A special application of DAF called the flotation filter was invented at the very end of the 1960s. It is a combination of flotation and rapid sand filtration, both of those being placed in the same tank. Flotation takes place in the upper part of the tank and the filter has been placed in the lower part of it. The direction of water flow is now vertically down from the free surface of water in the tank toward the deep-bed filter. This controls the direction of flow in the flotation space of the tank above the filter bed. The flow rate of water in flotation filters may be 10-15 m/h, but the flow conditions are still laminar. It is the threat that the head-loss of filters would grow too rapidly which in practice is limiting the hydraulic flow rate of flotation filters in this area. The third generation of DAF has been developed at the end of the 1990s. The operational idea is based on that of the flotation filter. The filter bed on the bottom of the tank has been replaced by a thin stiff plate with plenty of round orifices throughout the plate. This plate, having a very much lower flowing resistance than a sand filter can have, controls the vertical flow of water in the flotation space above the plate and distributes it evenly throughout the horizontal cross-section of the tank. The flotation tank is almost square seen from above and its depth is clearly more than the length and breadth of it. This kind of flotation unit can be operated with flow rates of water in the range 25-40 m/l. Even a flow rate of more than 60 m/h has been reported from this kind of DAF-units. There is no risk of clogging of the plate by suspended solids which could limit the flow rate. This is to say that it is possible to operate DAF also in turbulent flow conditions. The depth of the micro-bubble bed below the surface of water can be 1.5-2.5 m. There actually is a continuously regenerated micro-bubble bed in the tank filtering water which is going through this bed. The lower surface of the micro-bubble bed is really a horizontal one a little bit above the plate controlling the flow in the flotation space. The clarified water below the micro-bubble bed is totally clear. It can be said that in this case the removal of suspended solids takes place much more by filtering water by a deep-bed micro-bubble filter than by attaching micro-bubbles onto solids, when both of these are mixed with each other in the inlet shaft of the flotation unit, because the retention time of water in the inlet shaft is very short indeed. PMID- 11394262 TI - Removal of hydrocarbons from petrochemical wastewater by dissolved air flotation. AB - The dissolved air flotation (DAF) method has an important role in the removal of hydrocarbons, as well as in the protection of the biological treatment, which usually follows the DAF. The aims of this study were to evaluate the removal efficiencies of suspended solids, general organic matter, hydrocarbons and phenols by DAF, as influenced by the flocculant type, aluminum sulfate (alum) or a cationic polyelectrolyte. Laboratory batch experiments included chemical flocculation followed by DAF, controlling the flocculant dose and the air to solids ratio. The characterization of the influent and effluent was based on general analysis of organic matter (COD), suspended solids, hydrocarbons and phenols. The influent to all experiments was supplied daily from the outlet of a full scale oil-water gravitational separation unit at a petrochemical complex in Haifa, Israel. The influent contained hydrocarbons in the range of 20 to 77 mg/L. Usually less than 10% were found in "free" form, 70 to 80% were emulsified and 10 to 20% were dissolved. The DAF process enabled us to reduce the general hydrocarbon content by 50 to 90%. The effluent was characterized by stable and uniform levels of suspended solids, and oil, almost without depending on the influent concentrations. The results indicate that the chemical flocculation followed by DAF removed efficiently the emulsified phase, which could be aggregated and separated to the surface. However, it was found that the process could also remove substantial amounts of dissolved organic matter. This mechanism could be explained by the hydrophobic characteristics of some of the substances, which could bind to the solid surfaces. It was found that aggregates created by the flocculation with the cationic polyelectrolite (C-577) could remove up to 40% from the dissolved hydrocarbon. Alum flocs also indicated removal of soluble materials, mainly phenols. The results obtained in this study indicated the possibility to improve the protection of the biological treatment process by preliminary removal of hydrophobic compounds, usually considered as either inhibitory or toxic. This removal can be based on sorption onto aggregates created by chemical flocculation, which can be efficiently removed by dissolved air flotation. PMID- 11394263 TI - Dissolved air flotation treatment of concentrated fish farming wastewaters. AB - Fish farming wastewaters contain nutrients, phosphorus and nitrogen, which promote eutrophication in the typically shallow farming sites in Finland. Fish farming wastewater treatment is problematic because of large quantities of very dilute wastewater (200-600 m3/kg fish produced). In practice wastewater treatment is concentrated on suspended solids removal. Treatment can be done in two steps: concentration of the very dilute wastewater and subsequent treatment of the concentrated wastewater. Dissolved air flotation pilot trials were conducted using two types of concentrated wastewaters: settled solids from a sludge hopper of a cultivation basin and swirl separator concentrate. Two different pilot plants were used and performances compared. Both mechanical treatment and precipitation by ferric salts were applied. Depending on the influent quality, 70 to 90% phosphorus reductions were achieved without chemicals. Chemical precipitation and flotation produced 90% phosphorus reductions and effluent concentrations at the level of 0.05 mgP/l when 13 m3/(m2h) hydraulic loading was used. PMID- 11394265 TI - Pretreatment of bakery wastewater by coagulation-flocculation and dissolved air flotation. AB - The pretreatment of wastewater from a large-scale bakery was studied. In the coagulation-flocculation reaction, it was found that both alum and FeCl3 were effective in the jar tests. When at coagulant dosage of 90 to 100 mg/l, 55% of COD and 95 to 100% of SS could be removed. The optimum pH was at 6.0. In addition, the removal of SS was affected by pH more significantly, while the removal of COD was not affected in the pH range of 6.0 to 8.0. In the DAF experiments, 48.6% of COD and 69.8% of SS were removed in 10 min at a pressure of 4 kg/cm2, recycle ratio of 0.3 l/min, and pH of 6.0. Upon the addition of 100 mg/l of alum, the removal efficiency of COD did not increase while SS removal increased to 82.1%. It was found that 5-min flocculation time did improve the COD removal while it had little effect on SS removal. Flocculation for longer than 5 min did not enhance the flotation performance. Similar phenomena were observed when FeCl3 was used as the coagulant, except that flocculation had an insignificant effect on COD and SS removal. It was also found that FeCl3 was relatively more effective than alum. In summary, both coagulation-flocculation and DAF were efficient for the pretreatment. The advantages and disadvantages were discussed. PMID- 11394264 TI - Biosorptive flotation for metal ions recovery. AB - The solid/liquid separation of suspended, metals-loaded biomass was studied extensively by successfully applying flotation. Industrial non-living biomass samples of different origin have been tried in the laboratory as sorbents of metal ions, with the main stress on Streptomyces rimosus, an effective actinomyces. Flotation was used as the harvesting technique downstream, following the biosorption stage. Important parameters of the investigation were the solution pH, the surfactant type and concentration, the applied modification of biomass, etc. The reuse and recycling of biosorbent following elution was proved possible. As a result cleaned water was produced as underflow of the flotation process. PMID- 11394267 TI - Optimizing dissolved air flotation design and saturation. AB - Dissolved air flotation (DAF) of iron hydroxide precipitates at working pressures lower than 3 atm, using modified flotation units to improve the collection of fragile coagula, was studied. Conventional DAF flotation was studied as a function of saturation pressure in the absence and presence of surfactants in the saturator. Without surfactants, the minimum saturation pressure required for DAF to occur was found to be 3 atm. But, by lowering the air/water surface tension in the saturator, DAF was possible at a saturation pressure of 2 atm. This behavior was found to occur in both batch and pilot DAF operation tests and almost complete recovery of the precipitates was attained. Results are explained in terms of the minimum "energy" which has to be transferred to the liquid phase to form bubbles by a cavity phenomenon. Further, studies were conducted changing equipment design and feed bubbles size distribution (mixing micro and "mid-sized" bubbles). Thus, bubbles entrance position in the collision-adhesion zone ("capture" zone) was compared to bubble entrance position in the water flow inlet below the floating bed. A "mushroom" type diffuser was used for the "capture zone" experiment and better performance was obtained. Results are explained in terms of different mass transfer phenomena in the collection zone and in the separation zone. Finally, results obtained with the use of a column flotation cell working as normal DAF and with a wide bubble size range are presented. Results indicate good performance and some gains in process kinetics with middle size bubbles. PMID- 11394266 TI - Collision efficiency factor of bubble and particle (alpha bp) in DAF: theory and experimental verification. AB - The collision efficiency factor of bubble and particle (alpha bp) in dissolved air flotation (DAF) can be calculated theoretically by trajectory analysis, which takes into account both hydrodynamics and interparticle forces. To determine the theoretically optimum particle size for any given bubble size, a collision efficiency diagram for DAF was developed where collision efficiency is contoured on a plane of particle and bubble sizes for different conditions of particle zeta potential. A set of experiments tested the validity of the suggested collision efficiency diagram, and examined whether pretreatment is important and why slight coagulant overdosing and shorter flocculation times are generally preferred in DAF, both current accepted practice. Batch DAF reactors were used and kaolin samples were prepared from jar tests using different alum dosages and flocculation times. The particle size distribution, particle zeta potential, and turbidity removal in each experiment were measured, as were bubble size and zeta potential. The results agreed well with the predictions of the collision efficiency diagram and explained current practices. A collision efficiency diagram identifies the pretreatment goal, i.e., tailoring of the optimum characteristics required of particles (zeta potential and size) under existing operational bubble characteristic. PMID- 11394268 TI - A multiphase CFD model of DAF process. AB - A Eulerian-Eulerian multiphase CFD model is employed for the air/water flow. A 3D structure grid is used to incorporate the air nozzle and tank geometry. The fixed frictionless wall boundary approximating the free surface acts as a sink to allow the air bubbles to escape. The air/water volume fraction in the flotation tank is evaluated to determine the effective air/water fluid density. The floc particle is then introduced and is tracked in the air/water fluid using a disperse Lagrangean model. Fate of these flocs depends on their sizes and density. Flocs therefore can either escape through the top water surface, settles in the main tank or breakthrough under the outlet weir. The CFD model is developed for a full scale DAF tank to predict the flow dynamic, particle removal and settled solid profile. The general flow pattern is compared with flow visualisation using the underwater camera. Comparison of average fluid velocities is carried out using acoustic Doppler velocimetry ADV measurement. PMID- 11394269 TI - Modelling the global efficiency of dissolved air flotation. AB - The purpose of this paper is to examine how the efficiency of dissolved air flotation is affected by the size of bubbles and particles. The rise speed of bubble/particle agglomerates is modelled as a function of bubble and particle size, while the kinematics of the bubble attachment process is modelled using the population balance approach adopted by Matsui, Fukushi and Tambo. It is found that flotation, in general, is enhanced by the use of larger particles and larger bubbles. In particular, it is concluded that for the ultra-high surface loading rates of 25 m/hr or more planned for future flotation tanks, bubble size will have to be increased by a factor of two over the size currently employed in many facilities during dissolved air flotation. PMID- 11394270 TI - Small scale model for CFD validation in DAF application. AB - A laboratory model is used to measure the generic flow patterns in dissolved air flotation (DAF). The Perspex model used in this study allows the use of laser Doppler velocimetry (LDV), a non-invasive, high-resolution (+/- 2 mm s-1) laser technique of flow velocity measurement. Measurement of flow velocity in the single-phase situation was first carried out. Air-saturated water was then supplied to the tank and measurements of bubble velocity in the two-phase system were made. Vertical flow re-circulation was observed in the flotation zone. In the bottom of the flotation zone (near the riser) secondary flow re-circulation was observed, but only in the two-phase system. Another phenomenon was the apparent movement of flow across the tank width, which may be due to lateral dispersion of the bubble cloud. Data from preliminary computational fluid dynamics (CFD) models were compared against this measured data in the case of the single-phase system. The CFD model incorporating a k-epsilon model of turbulence was found to give closer agreement with the measured data than the corresponding laminar flow model. The measured velocity data will be used to verify two-phase computational fluid dynamics (CFD) models of DAF. PMID- 11394271 TI - Modelling of floc-bubble aggregate rise rates in dissolved air flotation. AB - The use of relatively simple, but conceptually sound mathematical models, is a powerful tool to identify and understand parameters that are critical to a process. In this paper, a model is presented which addresses the rise rate of floc-bubble aggregates in the DAF separation zone. The model uses Stoke's Law as a point of departure, which is then progressively extended to incorporate the non sphericity of the aggregates, the non-laminar nature of their movement, the fractal nature of the flocs, the physical constraints of attaching bubbles to a floc, and the limit on bubble numbers imposed by a typical air dosing system. The main findings are: There are two distinctly different DAF domains, namely a small floc domain and a large floc domain. In the small floc domain, the bubble size, the air volume, chemical dosing and the degree of flocculation have to be optimized and accurately controlled, while the large floc domain is less sensitive to these parameters. The calculated rise rates are significantly lower than some of the latest pilot testing results reported from the USA; a surprising finding at first. There are, however, numerous modelling simplifications that may explain this discrepancy. The most probable reasons are the further agglomeration of the aggregates in the separation zone due to differential rise rates or the recirculating flow within the white-water blanket, and the complex and poorly understood flow patterns within the separation zone. PMID- 11394272 TI - The flow structure in the separation zone of a DAF pilot plant and the relation with bubble concentration. AB - Fluid dynamics is believed to be of crucial importance for the removal mechanism in Dissolved Air Flotation. The paper is a presentation of experimental studies on velocity and air-content profiles in a DAF pilot plant. The correlation between air content and flow patterns is discussed. Two flow structures are detected. A stratified flow structure is defined by a horizontal transport of the water in a upper, less-dense layer, and a lower, downwards-vertical transport in the lower, dense layer. Between the layers a return flow is found. The short circuiting flow is defined as an immediate downward deviation towards the outlet arrangement at the bottom of the tank, probably causing decreasing effluent quality. Density gradients, caused by differences in air content in the tank, are believed to generate a stratification of the water body, thus causing the water to be transported in layers. Measurements of air-content profiles correlate to the measured flow structures, also indicating a separation of the water body in two layers with a distinct boundary. PMID- 11394273 TI - An integrated approach to dissolved air flotation. AB - DAFRapide techniques utilising reduced flocculation times, of the order of 5 minutes, together with flotation loading rates of up to 40 m/h, can be incorporated with conventional filtration systems operating at up to 20 m/h. Product quality of typically < 1 NTU and < 0.1 NTU after flotation and filtration respectively can be achieved. PMID- 11394274 TI - Experimental and modeling evaluation of upward and downward velocities in the coaxial flotation column. AB - This study was conducted to show the influence of upward velocity in the inner column and downward velocity in the outer column of the coaxial cylinder-type flotation column on the solids removal efficiency, solids concentration in the treated water, and so on. The SIMPLE (Semi-Implicit Method for Pressure Linked Equation) solution was applied to the coaxial flotation column to simulate the velocity vectors of the elements of water flowing in the column. The effects of solids loading and residence time in the agglomerate separation zone on the solids removal efficiency were also tested. In the pilot scale coaxial DAF column experiments with solids concentration of 1,000-2,000 mg of SS per liter and solids loading less than 350 kg/m2/day, approximately 90% of the solids removal efficiencies were obtained using the upward velocity of up to 110 cm/min in the contact zone of the inner column and the downward velocity of up to 30 cm/min in the outer column. In the simulation, similar results were observed as in the experiments. The solids loading in the excess of 350 kg/m2/day caused the instability of the sludge float layer and aggravated the quality of the treated water. PMID- 11394275 TI - Characteristics of alumino-humic flocs in relation to DAF performance. AB - This paper examines the implications of pretreatment in the context of the likely behaviour of flocs in DAF units. Evidence indicates that the maximum floc size (dmax) of alumino-humic is scaled by the Kolmogorov length (eta) with dmax/eta approximately 1. A larger ratio exists when polymer is present. Data from the literature suggests that velocity gradient in the inlet chamber may exceed 1000 s 1, suggesting eta approximately 40 microns. This view of the response of floc to shear is supported by shear tests carried out on samples of floc withdrawn from flocculators at full scale water treatment works. Using an underwater CCTV camera, coupled with image analysis, it is shown that floc sizes in conventionally designed flocculators are much bigger than the sizes which are likely to exist in DAF units. Circumstantial evidence gained from water treatment works suggests that DAF units perform satisfactorily whatever the size of the incoming floc. From such evidence, the authors question the conventional design of the flocculators in association with DAF units. PMID- 11394276 TI - A decade of large scale experience in dissolved air flotation. AB - In April 1990 Antwerpse Waterwerken brought a new DAF-plant into operation at the production centre Notmeir-Walem. The flotation unit, which has a capacity of 200,000 m3/d, was integrated in one of the two existing treatment lines. Its main objective was to eliminate the important raw water quality fluctuations caused by increasing algal growths. During the past ten years several optimisation studies were carried out which resulted in a further simplification of operation and mechanical design. This paper will focus mainly on the integration of dissolved air flotation in the entire treatment line, the consequences on the dual layer filtration and the operational experience and information gathered during the past decade. PMID- 11394277 TI - Using lessons learned and advanced methods to design a 1,500 Ml/day DAF water treatment plant. AB - The paper describes the method that led to the design of the 1,500 Ml/day dissolved air flotation (DAF) water treatment plant for Boston's water supply. In particular, the topics of flocculation techniques, floated solids removal and DAF recycle as they relate to very large capacity plant design are covered in detail. The use of mathematical models, including computational fluid dynamics (CFD) software, to refine the design is described. PMID- 11394278 TI - High-rate dissolved air flotation for water treatment. AB - This paper presents the results of an experimental investigation about the performance of a horizontal flow high-rate pilot scale Dissolved Air Flotation (HRDAF) unit containing inclined parallel plates for treating a coloured and low turbidity raw water. Experiments were performed with the DAF unit in order to verify the influence on flotation of: (i) the water velocity (Vh) between the plates, in the range 18 to 96.5 cm.min-1 with corresponding Reynolds numbers between 240 and 1060; (ii) the supplied air (S*) value ranging from 2.2 to 8.5 g of air/m3 of water; (iii) the angle of the plates (60 degrees or 70 degrees). The best pilot plant operational condition was obtained applying only 4.0 g/m3 (S*) with Vh around 18 cm.min-1 for treatment of water coagulated with a Al2(SO4)3 dosage of 40 mg.l-1. In these conditions, the unit presented very good removal efficiencies of colour (90%, residual of 10 uC), turbidity (88%, residual of 0.8 NTU) and TSS (94%, residual of 1.8 mg.l-1). Furthermore, the unit could operate at higher Vh values up to 76 cm.min-1 and still present good results. The DAF unit thus behaved as a high rate unit presenting good performance with low air requirement. PMID- 11394279 TI - Removal and fate of Cryptosporidium in dissolved air drinking water treatment plants. AB - In the first part of the paper, data from pilot plant studies are used to evaluate Cryptosporidium removal by dissolved air flotation (DAF) clarification and dual media filters under challenge conditions. Oocyst removals were investigated for design detention times and hydraulic loadings for winter and spring seasons. Coagulation was optimized for turbidity and removal of natural organic matter. DAF performance was better for spring water temperatures achieving 2.5 +/- 0.3 log removal of oocysts compared to 1.7 +/- 0.3 log removal in the winter. Cumulative log removal across DAF and filtration exceeded 5.4, and was not affected by water temperature. Low turbidities and particle counts are indicators of good treatment and good removals of Cryptosporidium. The second part of the paper uses a mathematical model to predict the fate of Cryptosporidium through a DAF plant and the impact of filter backwash recycle on oocyst build-up in the plant influent. Model predictions show that the fate of Cryptosporidium and the build-up of oocysts in the plant influent depend on: DAF performance, the percent of filtered water production used for backwashing, and the percent of filter backwash recycle flow. A DAF plant with 2.5% filtered water production for backwashing and that achieves 1.6 log removal or greater of oocysts by DAF clarification will not have a build-up of oocysts in the plant influent regardless of the recycle rate. This is because the oocysts are concentrated in the DAF floated sludge and not within granular filters. PMID- 11394280 TI - Treatment of spent filter backwash water using dissolved air flotation. AB - There is increasing interest in treating recovered spent filter backwash water in the drinking water industry. In the USA the Filter Backwash Recycling Rule will come into effect in the near future. The purpose of the Rule is to prevent the concentrated pathogenic agents, potentially in the filter backwash water, from being returned to the head of the water treatment works without some form of treatment or dilution. By treating this flow both public health and financial liability can be better managed by the operating utility. Dissolved Air Flotation (DAF) was investigated as a possible technology alternative to simple or advanced sedimentation techniques. This application is not widespread but sits somewhere in between the two normal applications of DAF as a high solids sludge thickener and a low turbidity clarification system. Given this a pilot plant program, supported by jar testing, was undertaken to determine the process capability and the design parameters for this application. DAF proved to be very suitable for backwash water recovery. DAF effluent turbidities of < 1.0 NTU could be easily obtained, when raw water turbidities were in excess of 50 NTU. Chemical requirements were low with only a single low dose of polymer required to bind the floc particles to form a solids matrix suitable for flotation. Flocculation contact times ranged from 0-10 minutes depending on the nature of the raw water. Recycle rates as low as 5% performed satisfactorily with no significant improvement when increased to 20%. Sludge solids of 3.5-9.6% dry solids were found and very low volumes of sludge, < 0.1% of the incoming flow make the DAF solids handling system very compact. PMID- 11394281 TI - Evaluation of the performance of full-scale packed saturators. AB - An evaluation of the performance of full-scale packed saturators was made. Measurements of saturator effluent air concentrations, saturator efficiency and precipitated are reported for both cold water and warm water operating conditions. Packed saturators were analyzed over a full range of operating pressures and loading rates. Measured results compared to predicted results based on a comprehensive model for the performance of packed saturators prove the model to be a useful tool in designing efficient and cost effective saturation systems. PMID- 11394282 TI - The use of dissolved air flotation in municipal wastewater treatment. AB - Flotation can be used in municipal wastewater treatment plants in different ways. Since the pollutants in wastewater to such a large extent are associated with particles, a very substantial treatment efficiency can be reached at a very small space, by using flotation in a chemical (or enhanced primary) treatment scheme. This is demonstrated in this paper with reference to results from small, prefabricated chemical plants based on flotation, which are frequently used in Norway. If used in connection with biological plants (for instance for nitrogen removal), the combination of biofilm reactors and flotation is especially advantageous because coagulation/flocculation/flotation can be placed directly after the bioreactor. Results from two such plants in Norway are presented. Recommendations with respect to design and operation of flotation plants in wastewater treatment are given. PMID- 11394283 TI - Evaluation of flocculation and dissolved air flotation as an advanced wastewater treatment. AB - A bench scale study was carried out in order to evaluate the applicability of dissolved air flotation (DAF) as an advanced treatment for effluents from three different domestic wastewater treatment processes, namely: (i) a tertiary activated sludge plant; (ii) an upflow sludge blanket anaerobic reactor (UASB); and (iii) a high-rate stabilization pond. PMID- 11394284 TI - Dissolved air flotation in drinking water production. AB - Dissolved Air Flotation (DAF) has become increasingly important in the field of potable water treatment, as a preferred option for treating upland and stored lowland waters. This paper outlines the development of dissolved air flotation (DAF) in potable water treatment, the benefits and disadvantages and the recent advances that has taken the process technology from an art to a science. PMID- 11394285 TI - Sewage treatment by anaerobic biological process associated with dissolved air flotation. AB - This paper presents the results of a study performed with a lab-scale batch DAF unit fed with previously coagulated (with FeCl3 or cationic polymer) effluent from a pilot scale up-flow anaerobic sludge blanket (UASB) reactor treating domestic sewage. The adequate coagulation/flocculation conditions--chemical dosage, time (Tf) and mean velocity gradient (Gf) in the flocculation step--and air requirements for flotation process were investigated. Best results were achieved for 65 mg.l-1 of FeCl3 at Tf around 15 min and Gf of 80 s-1. In the assays where only polymer was applied, 7 mg.l-1 of cationic polymer dosage gave optimum removals with Tf around 15 min and Gf of 30 s-1. Air requirements ranged from 9.5 to 19.0 g of air.m-3 wastewater. Best TSS (95% and residual of 2 mg.l 1), COD (85% and residual of 20 mg.l-1) and total phosphate (95% and residual of 0.6 mg.l-1) removals were obtained when applying FeCl3, although the use of cationic polymer also produced good level of TSS (74% and residual of 14 mg.l-1) and COD (75% and residual of 45 mg.l-1) removals. For the UASB-DAF (batch) system and FeCl3, global efficiencies would be 97.2% for COD, 97.9% for phosphate and 98.9% for TSS. PMID- 11394286 TI - Ozonation followed by coagulation/flocculation and flotation as post-treatment of the effluent from an anaerobic baffled reactor treating domestic sewage. AB - This paper reports on the use of ozonation and dissolved air flotation as a post treatment of the effluent from an anaerobic baffled reactor treating domestic sewage. After preliminary essays, the present experiment was performed fixing coagulant doses and, to all of them, some ozone doses were investigated. Later, the pH value and the ozone dose which provided the best removal efficiencies of all the parameters involved were tested, changing the coagulant dose and varying, for each of them, two ozone doses: the zero one and the optimum. Considering the best conditions of coagulation/flotation (ferric chloride dose of 65 mg.L-1 and pH around 5.5), the ozone dose application of 6 mg.L-1 led to a significant level in the removal efficiency of COD (80.4%), BOD (79.0%), total phosphate (93.4%), apparent color (91.9%) and turbidity (97.0%), demonstrating that the system seems to be efficient and capable of promoting a high degree of sewage post-treatment, reducing the coagulant dose until 30%, with a consequent reduction in the sludge generation. PMID- 11394287 TI - Bar coding objects with DNA. AB - A bioanalytical scheme is described for bar coding objects with DNA. Several DNA templates of different lengths are mixed together and the mixture used to label the object of interest. To disclose the bar code, the DNA mixture is eluted from the object, amplified by the polymerase chain reaction with a universal pair of primers, and then the amplified templates resolved by agarose gel electrophoresis. In its present state, the bar coding scheme allows for over a million different labels. The reliability of the scheme has been demonstrated in a typical situation by labelling a letter with a template mixture (approximately 1.5 fmol of DNA), sending the letter through the post and successfully reading the bar code on receipt of the letter. PMID- 11394288 TI - Investigation of water environments in a C18 bonded silica phase using 1H magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectroscopy. AB - High resolution 1H magic angle spinning (MAS) nuclear magnetic resonance (NMR) spectra have been obtained on typical C18 bonded silicas used in chromatographic solid-phase extraction separations. It has been shown for the first time that water molecules distributed in distinct physico-chemical environments within the chromatographic system can be detected directly using a simple 1H MAS NMR measurement. The resonances assigned to water protons in differing physico chemical environments have distinct chemical shifts, line widths, relaxation times (T1 and T2) and also exhibit temperature dependent coalescence behaviour. This novel MAS approach may lead to a better understanding of the environments of other analytes in mixtures during such separations. PMID- 11394289 TI - UV-detection capillary electrophoresis for benzo[a]pyrene and pyrene following a two-step concentration system using homogeneous liquid-liquid extraction and a sweeping method. AB - 10(-9) mol l-1 levels of polycyclic aromatic hydrocarbons (PAHs) suspected for certain noxious materials can be determined and separated by UV-detection capillary electrophoresis following a two-step concentration system. When the conditions of the homogeneous liquid-liquid extraction were [THF]T = 5 vol%, [HCl]T = 0.66 mol l-1, and [PFOA]T = 2 x 10(-3) mol l-1 (i.e., the volume of sample solution; 50 ml-->sedimented phase; 30 microliters), the extraction percentages of benzo[a]pyrene (Bap) and pyrene (Py) at 10(-6) mol l-1 were 102 and 97.5%, respectively. Also, when the total concentration factor(volume ratio), which includes that of the sweeping method, was 8335-fold, the determination range of Bap was 6.4 x 10(-9)-8.0 x 10(-7) mol l-1 and for Py, 8.0 x 10(-9)-7.0 x 10(-7) mol l-1. The detection limits (3 sigma) of Bap and Py were 1.6 x 10(-9) and 4.8 x 10(-9) mol l-1, respectively. PMID- 11394290 TI - Chemiluminescence of Ce(IV) and surfactant Tween 20. AB - It was found that cerium(IV) reacted with surfactant Tween 20 in acidic medium to generate green chemiluminescence. The maximum emission wavelength was about 478 nm. The enhancement and inhibition of the CL reaction by some polyphenols and anilines was studied and the possibility of analytical application was explored. PMID- 11394291 TI - A novel ion chromatographic method based on cation-exchange and acid-base interactions for the simultaneous determination of total alkalinity and monovalent cations in samples of microliter volume. AB - An ion chromatographic (IC) method based on the use of titrant (strong acid) as the stationary phase was developed for simultaneous determination of total alkalinity (TA) and monovalent cations. The titrant used in this study was obtained by initially loading lithium dodecylsulfate (Li-DS) onto a reversed phase material and then conditioning the column with a slightly acidified aqueous LiCl solution (a mixture of 50.0 mM LiCl and 0.1 mM H2SO4). When a small amount of a basic sample was injected onto a column prepared in this way, the basic species (Bn-) reacted predominantly with H+ on the stationary phase and the reaction with the eluent phase was negligible due to the very low concentration of eluent H+ (in the eluent, a molar ratio of [Li+]/[H+] = 250:1 applied). The stationary phase H+ consumed in the acid-base reaction was then re-supplied by H+ from the eluent. By monitoring the conductance of the eluent using conductivity, an induced peak resulting from the basic species was observed. Calibration graphs of peak areas vs. molar concentration of the basic species for OH-, HCO3- and H2PO4- were found to be identical. CO3(2-), HPO4(2-), and B4O7(2-) also gave identical calibration curves but their slope values were twice those for HCO3-. The detection limit for HCO3- was less than 3.2 microM and the calibration curve was linear up to 12.3 mM (injection volume, 100 microL). Seawater was directly analyzed and its total alkalinity was found to be 2.87 mM (RSD 0.53%, n = 5), which was in good agreement with the result of 2.88 mM (RSD 3.2%, n = 5) obtained using auto-potentiometric titration. Na+ and K+ were determined simultaneously and the concentrations were 481.6 and 10.6 mM, respectively. PMID- 11394292 TI - Nonlinear electrophoretic focusing of DNA macromolecules. AB - A new approach to focusing DNA molecules in a nonuniform electric field based on nonlinear mobility (L. L. Frumin, S. E. Peltek, S. Bukshpan, V. V. Chasovskikh and G. V. Zilberstein, PhysChemComm, 2000, 11) is proposed. The focusing is carried out in an alternating nonuniform electric field, created by using a wedge gel with hyperbolic boundaries. Methods of the theory of analytical functions were used to demonstrate that the fractions separated electrophoretically in such a wedge retain their rectilinear shape. Solutions for the focusing points for the case of mere velocity cubic nonlinearity were obtained as well as for the square velocity nonlinearity, suggested in a number of modern approaches. Gel electrophoresis experiments supported the possibility of a pronounced nonlinear focusing of DNA molecules. This nonlinear separation technique presents encouraging prospects for preparative isolation of long DNA fragments and development of new separation methods for bacterial fingerprinting. PMID- 11394293 TI - A fluorescent probe for monitoring nitric oxide production using a novel detection concept. AB - A fluorescent probe using a novel 'spin exchange' concept was developed for monitoring nitric oxide (NO) production. The probe is composed of 2,2,6,6 tetramethylpiperidine-N-oxyl (TEMPO) labeled with acridine and N dithiocarboxysarcosine (DTCS)-Fe(II) complex. When the non-fluorescent acridine TEMPO was incubated with DTCS-Fe(II) complex in buffer solution, the nitroxide radical in the acridine-TEMPO interacted with the Fe(II) through a redox interaction. This interaction recovered the fluorescence based on the acridine moiety. The addition of an NO-releasing reagent caused a fluorescent decrease of the probe due to the irreversible binding of NO to the Fe(II), and the amount of the fluorescent decrease strictly corresponded to that of released NO. Using this probe, less than 100 nM of NO can be detected. This probe system is not only useful for monitoring direct production of NO in an aqueous solution, but is also interesting as a basic concept by which to construct new types of NO fluorescent probes. PMID- 11394294 TI - Simultaneous ion-exclusion chromatography and cation-exchange chromatography of anions and cations in environmental water samples on a weakly acidic cation exchange resin by elution with pyridine-2,6-dicarboxylic acid. AB - A non-suppressed conductivity detection ion chromatographic method using a weakly acidic cation-exchange column (Tosoh TSKgel OApak-A) was developed for the simultaneous separation and determination of common inorganic anions (Cl-, NO3- and SO4(2-)) and cations (Na+, NH4+, K+, Mg2+ and Ca2+). A satisfactory separation of these anions and cations on the weakly acidic cation-exchange column was achieved in 25 min by elution with a mixture of 1.6 mmol L-1 pyridine 2,6-dicarboxylic acid and 8.0 mmol L-1 18-crown-6 at flow rate of 1.0 mL min-1. On this weakly acidic cation-exchange resin, anions were retained by an ion exclusion mechanism and cations by a cation-exchange mechanism. The linear range of the peak area calibration curves for all analytes were up to two orders of magnitude. The detection limits calculated at S/N = 3 ranged from 0.25 to 1.9 mumol L-1 for anions and cations. The ion-exclusion chromatography-cation exchange chromatography method developed in this work was successfully applied to the simultaneous determination of major inorganic anions and cations in rainwater, tap water and snow water samples. PMID- 11394295 TI - Copper fractionation by SEC-HPLC and ETAAS: study of breast milk and infant formulae whey used in lactation of full-term newborn infants. AB - This method will allow the determination of bound copper to low relative molecular mass compounds in milk. The milk whey obtained by ultracentrifugation was submitted to fractionation by size exclusion chromatography (SEC) on a TSK Gel2000 (Toso Haas) column with a mobile phase of 0.2 M NH4NO3 + NH3, pH 6.7. Fractions of effluent corresponding to the protein peaks were collected and the copper content was determined by ETAAS. The method was sensitive (LOD 0.4 microgram l-1 and LOQ 1.5 micrograms l-1 in the fraction; LOQ 7.5-22.5 micrograms l-1 referred to the milk sample and depended on fraction volume) and precise (RSD +/- 10%). Media sample recoveries from the column were 101.2%. Cu was predominantly present in fractions corresponding to relative molecular mass 76 and 15 kDa of breast milk while copper was mostly found in fractions corresponding to 14 and 38 kDa of cow's milk-based infant formulae; moreover, copper was eluted in the relative molecular mass region < 6 kDa. PMID- 11394296 TI - Development and optimisation of an improved derivatisation procedure for the determination of avermectins and milbemycins in bovine liver. AB - A robust procedure has been developed to overcome the instability problems experienced with the fluorescent derivative of eprinomectin. The procedure involves addition of acetic acid, together with the typical reagents methylimidazole and trifluoroacetic anhydride, to produce a fluorescent molecule that can be determined by high performance liquid chromatography (HPLC) with fluorescence detection. Derivatisation is completed in 30 min at 65 degrees C. This derivatisation procedure was shown to be suitable, also, for the related compounds, moxidectin, abamectin, doramectin and ivermectin. A multi-residue method for these compounds in bovine liver has been developed using the derivatisation procedure. Samples are extracted with acetonitrile; followed by clean-up on deactivated alumina and C18 solid phase extraction (SPE) cartridges. The method was validated using bovine liver fortified at levels of 4 and 20 micrograms kg-1 with the drugs. The mean recovery ranged between 73 and 97%. The intra- and inter-assay variations showed relative standard deviations typically of < 6% and < 14%, respectively. The limit of quantitation of the method is 2 micrograms kg-1 (ppb). PMID- 11394297 TI - Sensitive determination of methylenedioxylated amphetamines by liquid chromatography. AB - Different strategies for the liquid chromatographic determination of methylenedioxylated amphetamines were evaluated: separation and detection of underivatized analytes by (i) UV or (ii) fluorescence, (iii) derivatization with 3,5-dinitrobenzoyl chloride followed by separation and UV detection of the derivatives formed and (iv) derivatization with 9-fluorenylmethyl chloroformate (FMOC) and subsequent separation and fluorimetric detection of the derivatives. The compounds tested were 3,4-methylenedioxyamphetamine (MDA), 3,4 methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyethylamphetamine (MDE). On the basis of these studies, a new procedure for the chromatographic determination of MDA, MDMA and MDE is proposed, based on derivatization with FMOC. The described procedure allows the quantification of the tested compounds with adequate linearity, reproducibility and accuracy in the concentration interval 0.5-20.0 micrograms mL-1. The limits of detection were 0.01 microgram mL 1 for MDA and 0.025 microgram mL-1 for MDMA and MDE. The utility of the described assay was tested by determining methylenedioxylated amphetamines in plasma and urine. PMID- 11394298 TI - Automated measurement system for H2O2 in the atmosphere by diffusion scrubber sampling and HPLC analysis of Ti(IV)-PAR-H2O2 complex. AB - An automated selective measurement system for monitoring hydrogen peroxide (H2O2) in the atmosphere was developed (using a diffusion scrubber coupled to a high performance liquid chromatography) due to the importance of H2O2 in understanding tropospheric chemistry and its harmful effects on vegetation. H2O2 in the atmosphere was effectively collected by a diffusion scrubber, which consisted of a hydrophobic porous polytetrafluorethylene (PTFE) tube positioned concentrically within a Pyrex glass tube. Titanium(IV)-4-(2-pyridylazo)resorcinol (Ti(IV)-PAR) solution acidified at pH 2.2 was used as the scrubbing solution for the diffusion scrubber. After the collection of the air sample by the diffusion scrubber, the pH value of the Ti(IV)-PAR scrubbing solution was adjusted to pH 11.9 to form a stable complex of Ti(IV)-PAR-H2O2. An aliquot of the sample solution was injected into a high performance liquid chromatograph equipped with a semi-micro-reversed phase column and a spectrophotometric detector set at 508 nm for separating and determining the Ti(IV)-PAR-H2O2 complex. The automated measurement could be performed at 60 min intervals. The collection efficiency of H2O2 was higher than 98% at an air flow rate of 1.0 l min-1. The detection limit (3 sigma of the blank value) of H2O2 was 9 parts per trillion by volume (pptv) for an air sampling volume of 55.1. The interference from coexisting O3 or SO2 in the atmosphere was negligible during the collection of H2O2 by the diffusion scrubber. The developed automated measurement system was suitable for monitoring H2O2 in the atmosphere. PMID- 11394299 TI - Application of central composite designs for optimisation of the chromatographic separation of monomethylarsonate and dimethylarsinate and of selenomethionine and selenite by ion-pair chromatography coupled with plasma mass spectrometric detection. AB - Central composite designs (CCDs) were used in the study of the ion-pair chromatographic separation of arsenic and selenium species in tap water: monomethylarsonate, dimethylarsinate, selenomethionine and selenite. The ternary eluent consisted of tetrabutylammonium phosphate (TBA), sodium hydrogenphosphate (Na2HPO4) and 1% acetonitrile. CCD allowed the investigation of the influence of the eluent parameters, which varied from 0.5 to 4.2 mmol l-1 Na2HPO4, 0.5 to 4.2 mmol l-1 TBA and pH 4.9 to 8.2, on the capacity factors (k') of arsenic and selenium compounds. Furthermore, another mathematical model that permitted the variation of the chromatographic selectivity of species, computed from their retention data to be followed, was investigated. This showed the ability to locate the optimum conditions within the experimental design, so that arsenic and selenium species could be simultaneously quantified with good efficiency and resolution. A comparison between the predicted and the experimental response values was made in order to assess the prediction quality of the model. Response surfaces and isoresponse curves obtained from the mathematical models allowed the determination of the optimum chromatographic conditions and the robustness of the method. The predicted optimum zone allowing satisfactory determination of both arsenic and selenium compounds was pH 5.5-6.5, 2.5 mmol l-1 Na2HPO4 and 3.0-4.0 mmol l-1 TBA. PMID- 11394300 TI - Automated on-line in-tube solid-phase microextraction followed by liquid chromatography/electrospray ionization-mass spectrometry for the determination of chlorinated phenoxy acid herbicides in environmental waters. AB - A method for the determination of six chlorinated phenoxy acid herbicides in river water was developed using in-tube solid-phase microextraction (SPME) followed by liquid chromatography/electrospray ionization-mass spectrometry (LC/ESI-MS). In-tube SPME is an extraction technique for organic compounds in aqueous samples, in which analytes are extracted from a sample directly into an open tubular capillary by repeated draw/eject cycles of the sample solution. Simple mass spectra with strong signals corresponding to [M-H]- and [M-RCOOH]- were observed for all herbicides tested in this study. The best separation of these compounds was obtained with a C18 column using linear gradient elution with a mobile phase of acetonitrile-water containing 5 mmol l-1 dibutylamine acetate (DBA). To optimize the extraction of herbicides, several in-tube SPME parameters were examined. The optimum extraction conditions were 25 draw/eject cycles of 30 microliters of sample in 0.2% formic acid (pH 2) at a flow rate of 200 microliters min-1 using a DB-WAX capillary. The herbicides extracted by the capillary were easily desorbed by 10 microliters acetonitrile. Using in-tube SPME LC/ESI-MS with time-scheduled selected ion monitoring, the calibration curves of herbicides were linear in the range 0.05-50 ng ml-1 with correlation coefficients above 0.999. This method was successfully applied to the analysis of river water samples without interference peaks. The limit of quantification was in the range 0.02-0.06 ng ml-1 and the limit of detection (S/N = 3) was in the range 0.005 0.03 ng ml-1. The repeatability and reproducibility were in the range 2.5-4.1% and 6.2-9.1%, respectively. PMID- 11394301 TI - Determination of linezolid in human plasma by LC-MS-MS. AB - A rapid, sensitive and selective LC-atmospheric pressure-chemical ionization-MS MS method for the determination of the new antimicrobial agent, linezolid, in human plasma using selected reaction monitoring (SRM) was developed. Linezolid and the internal standard were extracted from the biological samples by solid phase extraction (SPE) and analyzed on a reversed-phase Shim Pack CLC-CN, C18 column with the mobile phase of acetonitrile and 20 mM ammonium acetate solution (4 + 1 v/v). Detection was accomplished using an LCQ mass spectrometer (Finnigan), which was programmed in positive MS-MS mode to permit measurement of the fragment ions of linezolid and internal standard at m/z 296.2 and 223.2, respectively. The assay run-time was less than 3.5 min. Quantitative analysis was based on peak area ratio of linezolid to the internal standard. Calibration plots were established over the concentration range of 0.1-20 micrograms ml-1 of linezolid with the lowest detection limit of 0.05 microgram ml-1 using 10 microliters sample volume. The SPE technique quantitatively recovered linezolid and the internal standard from the plasma samples at a percentage range of 89.1 93.7%. Determination of control samples of linezolid in plasma validated the LC MS-MS-SRM method. Intra-assay and inter-assay precision were in the range of 5.1 11.4% relative standard deviation, whereas the intra- and inter-accuracy were in the range of 97.5-114.0% of the nominal concentrations of linezolid added. The data confirmed that the plasma samples of linezolid were stable at room temperature and when stored at -20 degrees C for at least 10 d. The developed LC MS-MS-SRM method is recommended for the determination of linezolid in human plasma. PMID- 11394302 TI - Analysis of badger urine volatiles using gas chromatography-mass spectrometry and pattern recognition techniques. AB - The potential for badger urine to signal olfactory information relating to sex, age class and seasonality was investigated by performing GCMS headspace analysis followed by pattern recognition statistical analysis on 84 urine samples collected from different categories of animal. Approximately 300 compounds were identified using library searching, and GCMS peak areas were recorded for the 33 most common. PCA was performed on the normalised and standardised data from all badgers, through which significant seasonal trends and groupings of homologous series of compounds were detected. PCA was also performed on the three subgroups of adults in the spring, summer and autumn, and a level of sexual discrimination was possible during the latter two seasons. Malanobis distances on the scores of the first five principal components provided good discrimination for these three subgroups, but discrimination was poor when all samples were analysed together. This, combined with the initial results of the PCA, confirms that a strong seasonal trend is imposed upon the sexual trend in this dataset. Our initial analysis indicates that badger urine potentially contains olfactory cues relating to sex and season. The relevance of these findings to understanding olfactory communication in mammals is discussed. PMID- 11394303 TI - Determination of phytochelatins by capillary zone electrophoresis with electrospray tandem mass spectrometry detection (CZE-ES MS/MS). AB - The coupling of capillary zone electrophoresis with electrospray mass spectrometry was optimized for the direct determination of phytochelatins (PCs) in extracts obtained from cells and plants that had been exposed to metal stress. Gluthathione and phytochelatins belonging to the different families (gamma Glu Cys)nGly (n-PC), (gamma Glu-Cys)nSer, (gamma Glu-Cys)n beta Ala and (gamma Glu Cys)n were separated in an uncoated capillary at pH 4 using a 5 mM ammonium acetate buffer, and detected by electrospray (ES) MS in the full scan mode (300 1100 u). The use of on-line tandem MS detection in the product ion scan mode of putative protonated molecules of PCs allowed the unambiguous confirmation of the identity of the compounds detected by ES MS. The operational conditions were optimized and the figures of merit were evaluated using n-PC2, n-PC3 and n-PC4 standards purified from a mixture obtained after the reaction of glutathione in the presence of Cd2+ and the enzyme PC-synthase. The method was applied to the characterization of bioinduced ligands in cell cultures of soybeans (Glycine max) and in rice (Oryza sativa) roots without the need for a preliminary sample cleanup by size-exclusion and/or reversed phase chromatography. PMID- 11394304 TI - Determination of U-236 in sediment samples by accelerator mass spectrometry. AB - 236U is produced only by neutron irradiation of uranium and therefore is potentially useful as a marker for anthropogenic uranium in the environment. Accelerator mass spectrometry (AMS) provides a technique for the determination of very low concentrations of actinide nuclides, and has now been applied to the determination of 236U:235U ratios in an intertidal sediment core collected from the North Irish Sea. Combining measurements of the 238U mass concentrations calculated from alpha spectrometry with 238U:235U ratios from ICP-MS and 236U:235U ratios from AMS has allowed the estimation of the mass concentrations of 236U in the sediments. 236U mass concentrations are in the range 10(-8) to 10( 9) g kg-1, and 236U:238U atom ratios in the range from 10(-5) to 10(-6), well above natural baseline levels. Uncertainties based on propagation of measurement errors were less than +/- 10% although +/- 15% is perhaps a more realistic estimate of overall uncertainty. PMID- 11394305 TI - A method for the measurement of catechol-O-methyltransferase activity using norepinephrine, an endogenous substrate. AB - We propose a highly sensitive method for the measurement of catechol-O methyltransferase (COMT) activity with norepinephrine (NE), an endogenous native substrate. The product, normetanephrine, was determined by high-performance liquid chromatography (HPLC)-peroxyoxalate chemiluminescence reaction detection or, if required, less sensitive fluorescence detection. For the measurement of membrane-bound (MB)-COMT activity in the rat erythrocyte, the HPLC-peroxyoxalate chemiluminescence reaction detection was employed. Soluble (S)- and MB-COMT activities in the rat erythrocyte were 22.9 +/- 2.5 and 4.62 +/- 1.23 pmol min-1 (mg protein)-1, respectively (n = 5). The Km values obtained for S- and MB-COMT were 366 +/- 31 mumol l-1 and 12.0 +/- 1.1 mumol l-1, respectively (n = 5), suggesting that the use of NE as a substrate would give more precise information on the role of both isoenzymes. However, with dihydroxybenzoic acid as an artificial substrate, the Km values for S- and MB-COMT were similar, with values of 69.2 +/- 11.4 mumol l-1 and 72.2 +/- 9.2 mumol l-1, respectively. The proposed method is thought to be useful for the measurement of both S-COMT and MB-COMT activities, and would give us critical information on the role of metabolism of catecholamines in rat tissues. PMID- 11394306 TI - A microbiological six-plate method for the identification of certain antibiotic groups in incurred kidney and muscle samples. AB - A microbiological method was developed for group level identification of antibiotics in incurred kidney and muscle samples from cattle and pigs. The method was composed of six test bacterium-plate growth medium combinations and the result was recorded as a profile of growth inhibition zones. The sample profiles were compared to two sets of references: one constructed with standard antibiotic solution profiles, and the other with these combined with profiles of microbiologically and chemically identified residues from incurred samples. The algorithm employed in profile comparison located the minimal sum of absolute pairwise differences over the tests, with the addition of a number of experimentally observed intra-test criteria. Chemical identification and quantitation of incurred residues was based on liquid chromatography. The method identified penicillin G as a penicillinase sensitive penicillin, enrofloxacin and ciprofloxacin belonging to fluoroquinolone group, and oxytetracycline belonging to tetracycline group. Each of these residues was microbiologically identified below the Maximum Residue Limit (MRL) for kidney tissue. Combining sample profiles with the standard reference data set did not enhance the resolution. Microbiological and chemical identification test results were in good agreement. The results of this study show that a microbiological identification method is a useful tool in preliminary characterisation of antibiotic residues in animal tissues. PMID- 11394308 TI - An integrated relay/nitrate reductase field-effect transistor for the sensing of nitrate (NO3-). AB - An integrated enzyme-functionalized field-effect transistor (ENFET) device for the sensing of nitrate ions is described. An aminosiloxane-functionalized gate interface is modified with N-methyl-N'-(carboxyalkyl)-4,4'-bipyridinium relay units. The complex formed between nitrate reductase and the bipyridinium units on the gate surface is crosslinked with glutaric dialdehyde to yield a stable relay enzyme layer on the gate interface. In the presence of sodium dithionite as electron donor, the biocatalyzed reduction of nitrate to nitrite ion is stimulated. The ratio between the oxidized and reduced states of the bipyridinium sites regulates the gate potential, and is controlled by the concentration of NO3 ions in the system. The effect of the chain length tethering the N-methyl-N' (carboxyalkyl)-4,4'-bipyridinium units to the gate surface on the biocatalyzed reduction of NO3- ions, and on the NO3- FET sensor performance is discussed. The devices that include the bipyridinium units tethered to the gate interface with methylene chain length, -(CH2)n, where n > or = 7, reveal a detection limit of 7 x 10(-5) M for nitrate and a sensitivity of 52 +/- 2 mV dec-1. The response time of the device is as low as 50 s, and the operational time of the system is ca. 85 s. We estimate the surface coverage of nitrate reductase on the gate surface to be ca. 1.2 x 10(-12) mol cm-2. PMID- 11394307 TI - FTIR sensing of inhomogeneous systems using immobilized organometalcarbonyl probe groups. AB - Tricarbonyl(eta 5-cyclohexadienyl)iron(0) and dicarbonyl(triphenylphosphine)(eta 5-cyclo-hexadienyl)iron(0) were derivatized by attachment of an aminopropylsilyl link and covalently attached to fumed silica particles. The fumed silica was coated onto the ZnSe element of an attenuated total reflection (ATR) cell for Fourier transform infrared (FTIR) spectroscopic analysis. The immobilized organometalcarbonyl probe groups are shown to retain their capacity to function as a key element of a molecular sensor assembly and the nu(CO) bands of the two probe groups were interrogated to calibrate the responses for 0-5% levels of dodecane in cyclohexanol to within +/- 0.1%. The potential for dual sensing is described and the simultaneous monitoring of two discrete regions of a dynamically varying inhomogeneous system is reported for the determination of dodecane in cyclohexanol as solutions mix across a permeable barrier in the ATR cell. PMID- 11394309 TI - Integration of microfabricated needle-type glucose sensor devices with a novel thin-film Ag/AgCl electrode and plasma-polymerized thin film: mass production techniques. AB - We developed an integrated array of needle-type biosensors employing a novel process of fabrication, comprising conventional semiconductor fabrication and micromachining technology. Amperometric sensing electrodes with plasma polymerized films and a thin-film Ag/AgCl reference electrode were directly integrated on a glass substrate with thin-film process, e.g., sputtering. An enzyme was immobilized on the electrode via the plasma-polymerized film, which was deposited directly on the substrate using a dry process. The novel thin-film Ag/AgCl reference electrode showed stable potentials in concentrated chloride solutions for a long period. The plasma-polymerized film is considered to play an important role as an interfacial design between the sensing electrode and the immobilized enzyme considering that the film is extremely thin, adheres well to the substrate (electrode) and has a highly cross-linked network structure and functional groups, such as amino groups. The results showed increments of the sensor signal, probably because the plasma-polymerized film allowed a large amount of enzyme to be immobilized. The greatest advantage is that the process can permit the mass production of high-quality biosensors at a low cost. PMID- 11394310 TI - A piezoelectric biomimetic sensor for aminopyrine with a molecularly imprinted polymer coating. AB - A molecularly imprinted polymer for aminopyrine was synthesized using methacrylic acid as functional monomer. The polymer was employed as the recognition element of a piezoelectric bulk acoustic wave biomimetic sensor for aminopyrine. Influencing factors were investigated in detail and optimized. This sensor exhibited high selectivity and sensitivity to aminopyrine. The response range of the sensor was between 5.0 x 10(-8) and 1.0 x 10(-4) M with a detection limit of 2.5 x 10(-8) M in the aqueous system. Scatchard analysis with UV spectrophotometry showed that the same class of binding sites was formed in the molecularly imprinted polymer in the studied concentration range, and the dissociation constant and the apparent maximum number of these binding sites were estimated to be 2.29 mM and 165.0 mumol g-1 dry polymer, respectively. Impedance analysis was employed to verify the imprinting effect and lack of variation in the viscoelasticity of the polymer coating during detection. PMID- 11394311 TI - Determination of sulfur dioxide in vitriol plant wastewater by using a polyNiMe4TAA electrochemically modified Pt microelectrode. AB - A Pt microelectrode modified with nickel(II) polytetramethyldibenzo[b,i]tetraaza[14]annulene was prepared by electropolymerization of nickel(II) tetramethyldibenzo[b,i]tetraaza[14]annulene monomers and applied to determine sulfur dioxide in vitriol plant wastewater. For determination of SO2 with this electrochemically modified Pt microelectrode, the linear range was from 9.6 x 10(-6) to 2.4 x 10(-4) mol L-1, the sensitivity was 1.8 x 10(-4) A L mol-1, the detection limit was calculated to be 4.8 x 10(-6) mol L-1 (S/N = 3), the response time was less than 20 s and the relative standard deviation was found to be 2.1% on analyzing 4.8 x 10(-5) mol L-1 SO2 solution repeatedly (n = 7). These results demonstrated good accuracy compared with those obtained by the conventional iodimetric method. PMID- 11394312 TI - Capillary-scale polarimetry for flowing streams. AB - A micro-polarimeter with a 40 nL probe volume was configured so that it is compatible with capillary-scale flowing stream analysis. The optical configuration consists of two polarizing optics, a capillary, a laser source and a photodetector which is very simple to configure with low cost components. This unique polarimeter is based upon the interaction of a linearly polarized laser beam and a capillary tube, in this case one with an inner diameter of 250 microns. Side illumination of the tube results in a 360 degrees fan of scattered light, which contains a set of interference fringes that change in response to optically active solutes. Solutes that exhibit optical activity are quantifiable and are detected by analyzing the polarization state of the backscattered light. The ability of the instrument to make extremely sensitive optical activity measurements in flowing streams is shown by the determination of (R)-mandelic acid, with a detection limit of 66 x 10(-6) M (507 x 10(-12) g), and the non optically active control, glycerol. Additionally, the detector was configured to minimize refractive index perturbations. PMID- 11394313 TI - Differential-pulse voltammetric determination of trace formaldehyde using magnetic microspheres and magnetic electrode. AB - A new type of magnetic polymer microsphere containing acylhydrazine groups on the surface was synthesized. They can be reacted with formaldehyde to produce an electroactive adduct. Reduction of these derivatives following aggregation on a magnetic electrode is possible and is effective in the indirect determination of formaldehyde. The experimental conditions and electrode structure are discussed. Under the optimum conditions, it was found that the peak potential (Ep) of formaldehyde is -1.01 V (vs. Ag/AgCl). Formaldehyde in the range 1-1000 micrograms l-1 can be determined. The detection limit for formaldehyde is 0.3 microgram l-1 and the relative standard deviation for the determination of 100 micrograms l-1 formaldehyde was 2.26%. The method was applied to the determination of formaldehyde in environmental samples with satisfactory results. PMID- 11394314 TI - Investigation of heroin profiling using trace organic impurities. AB - The acidic and neutral impurities in heroin samples (46 Purified South-East Asian (PSEA) and 8 non-PSEA samples) were analysed using gas chromatography. Mass spectral detection allowed the construction of a comprehensive library of over 649 impurities. A variety of statistical procedures were used to select 70 viable profiling parameters. Cluster analyses were used to investigate the similarities between samples using organic parameters. A blended PSEA profile was constructed for comparison with samples of unknown origins. Resolution between samples of PSEA/non-PSEA origins was demonstrated with a selection of 25 (18 continuous and 7 dichotomised) discriminating factors. Several predictive logistic models were created using up to 18 parameters, explaining 85-100% of the variation in the experimental data. PMID- 11394315 TI - Error-compensated kinetic determinations by detecting the reagent in successive reactions. AB - An error-compensated algorithm based on detecting the reagent in successive reactions is proposed. The algorithm was first studied by using simulated data and it was found that it performed well in compensating for the system error and eliminating random noise. Then the algorithm was examined by applying it to the determination of epinephrine and norepinephrine under different experimental conditions. The results revealed that the algorithm compensated errors produced not only by the changes of rate constants, but also by the variation of equilibrium signals. PMID- 11394316 TI - Sensitive detection of tetracyclines using europium-sensitized fluorescence with EDTA as co-ligand and cetyltrimethylammonium chloride as surfactant. AB - The determination of tetracyclines (TC) in aqueous solutions, based on europium sensitized fluorescence, has been improved using EDTA as co-ligand and cetyltrimethylammonium chloride as surfactant. The method involves working in slightly alkaline solutions with the formation of a new chelate where the lanthanide ion is bound to the beta-diketone group. The method is about 6 times more sensitive than that with the Eu-TC-Triton system and LODs are 2.5 x 10(-10), 5 x 10(-10), 1.5 x 10(-9) and 2 x 10(-9) mol l-1 for TC, oxytetracycline, chlortetracycline and doxycycline, respectively. The method has been applied to the determination of TC in calf serum without sample pretreatment. The mean recovery was close to 102% and the lowest concentration attainable in serum samples was better than 0.1 microgram ml-1. PMID- 11394317 TI - Improved fluorimetric determination of dissolved aluminium by micelle-enhanced lumogallion complex in natural waters. AB - A modification of the aluminium-lumogallion fluorescence measurement in the presence of the non-ionic surfactant Triton X-100 is presented. The detection limit for dissolved Al is 0.7 nM, with a relative standard deviation of 3.6% at an Al level of 5.0 nM. Compared with previously reported methods in the literature, the method described here is free from matrix effects and can be used for the determination of aluminium in fresh, estuarine and saline waters. The interferences from iron and fluoride were minimized by the addition of o phenanthroline and Be2+, respectively. The analysis of NIST SRM 1643C and PRC standard 2430101 by the proposed method provides results consistent with the certified values. A successful inter-laboratory calibration exercise also demonstrates the merit of the proposed method for the determination of Al in environmental and marine sciences. PMID- 11394318 TI - Homogeneous liquid-liquid extraction method for the selective spectrofluorimetric determination of trace amounts of tryptophan. AB - Twenty-one amino acids were derivatized with fluorescamine (FLA) under basic conditions (pH 9) and the extraction of the amino acid-FLA derivatives was investigated using a homogeneous liquid-liquid extraction with perfluorooctanoic acid (HPFOA) based on phase separation under strongly acidic conditions. Under the optimum concentration conditions for the reagents ([PFOA]T = 3 x 10(-3) mol dm-3, [acetone]T = 3 vol.%, [HCl]T = 1.8 mol dm-3), the concentration factor was approximately 1000-fold (i.e., 30 microliters of the sedimented liquid phase was produced from 33 ml of the homogeneous aqueous solution). The percentage extraction (E) was determined for the 21 amino acid-FLA derivatives; the value for the tryptophan (Trp)-FLA derivative was 80.9%, whereas the other derivatives were not almost extracted (E < 0.4%). The Trp-FLA derivative was selective for the extraction using the homogeneous liquid-liquid extraction method with HPFOA. After the sedimented liquid phase containing Trp-FLA has been placed on a polytetrafluoroethylene filter-paper, the fluorescence intensity was determined using a spectrofluorimeter with filter-paper as the solid-sample holder. The calibration graph of Trp was linear over the range 1.0 x 10(-8)-1.5 x 10(-6) mol dm-3. The relative standard deviation for the central value of the calibration graph was 4.5% (five determinations) and the detection limit (S/N = 3) was 8.9 x 10(-9) mol dm-3. When the proposed method was applied to the highly sensitive spectrofluorimetric determination of Trp in animalin-L syrup, the results were satisfactory. PMID- 11394319 TI - Study of the reaction of proteins with Beryllon II-AlIII by the Rayleigh light scattering technique and its application. AB - The determination of proteins with tetrasodium 2-(3,6-disulfo-8 hydroxynaphthylazo)-1,8-dihydroxynaphthalene-3,6-disulfonate (Beryllon II) by Rayleigh light scattering (RLS) was studied. The weak RLS of the Beryllon II bovine serum albumin (BSA) complex can be greatly enhanced by the addition of Al3+ in the pH range 5.6-7.2; there was a maximum RLS platform at 400-420 nm. Based on the reaction between Beryllon II, Al3+ and proteins, a new method for the determination of proteins was developed. This method is very sensitive [0.20 41.42 micrograms ml-1 for BSA and 0.18-48.15 micrograms ml-1 for human serum albumen (HSA)], rapid (< 2 min), simple (one step) and tolerant towards most interfering substances. The effects of different surfactants were also examined. Four samples of protein in human serum were determined; the maximum relative error was no more than 5% and the recovery was 96-105%. PMID- 11394320 TI - The influence of chlorine on the intensity of metal atomic lines emitted by an electrolyte cathode atmospheric glow discharge. AB - The effect of different matrix anions in the solution on the intensity of metal atomic lines was investigated. A significant increase in intensity was found for chloride anions compared with nitrate and sulfate anions. This effect was even greater when the appropriate acids were applied. A further enhancement of the metal line intensities could be observed when HCl was used in the solution phase and simultaneously elemental chlorine was mixed with atmospheric air at levels up to 6-10 vol.%. This double effect was especially high for the Cu, Ni and Pb resonant atomic lines at higher chlorine-to-air ratios in the gas phase, and the W-anode tip was destroyed by chemical burning. The application of volatile organic chlorine compounds (carbon tetrachloride and chloroform) in the gas phase, even without any acidification, also caused an enhancement of the metal line intensities. The experimental results can be attributed to the different rates of the ion-ion (positive metal ion-negative chloride ion) and the positive metal ion-electron recombination processes taking place in the cathode dark space of the discharge plasma, yielding neutral metal atoms for excitation. This study is important for the on-line measurement of heavy metals in liquids. PMID- 11394321 TI - Speciation and preconcentration of vanadium(V) and vanadium(IV) in water samples by flow injection-inductively coupled plasma optical emission spectrometry and ultrasonic nebulization. AB - An on-line separation, preconcentration and determination system for vanadium(IV) and vanadium(V) comprising inductively coupled plasma optical emission spectrometry (ICP-OES) coupled to a flow injection (FI) method with an ultrasonic nebulization (USN) system was studied. The vanadium species were retained on an Amberlite XAD-7 resin as a vanadium-2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (V-5-Br-PADAP) complex at pH 3.7. Enhanced selectivity was obtained with the combined use of the formation on-line of the complexes and 1,2 cyclohexanediaminetetraacetic acid (CDTA) as masking agent. The vanadium complexes were removed from the microcolumn with 25% v/v nitric acid. A sensitivity enhancement factor of 225 was obtained with respect to ICP-OES using pneumatic nebulization (15-fold for USN and 15-fold for the microcolumn). The detection limit for the preconcentration of 10 mL of aqueous solution was 19 ng L 1. The precision for 10 replicate determinations at the 5 micrograms L-1 V level was 2.3% relative standard deviation (RSD), calculated from the peak heights obtained. The calibration graph using the separation and preconcentration system for vanadium species was linear with a correlation coefficient of 0.9992 at levels from near the detection limits up to at least 100 micrograms L-1. The method was successfully applied to the speciation of vanadium in river water samples. PMID- 11394322 TI - Determination of airborne formaldehyde by active sampling on 3-methyl-2 benzothiazolinone hydrazone hydrochloride-coated glass fibre filters. AB - Formaldehyde was sampled with the use of a standard miniature glass fibre filter coated with 3-methyl-2-benzothiazolinone hydrazone hydrochloride (MBTH). The formaldehyde hydrazone formed [i.e., the adduct of formaldehyde (HCHO) and MBTH] was desorbed from the filter with water and then oxidised by an iron(III) chloride-sulfamic acid solution to form a blue cationic dye in acidic medium which was subsequently determined by visible absorption at 628 nm. The recovery of HCHO as the cationic dye from MBTH-coated filters is 87-102% in the range 0.065-2.9 micrograms of HCHO. This corresponds to 4.3-193.3 micrograms m-3 in a 15 L air sample. The collection efficiency of the MBTH-coated filter is higher than 90%. When the filter sampling system is used in active mode, air can be sampled at a rate of up to 1 L min-1, affording an overall sensitivity of about 3 micrograms m-3, corresponding to about 2 ppb v/v HCHO at 1 atm and 273 K. The method was successfully applied to the determination of HCHO in samples of indoor and outdoor air with satisfactory results. PMID- 11394324 TI - ECG of the month. Reality check. Complicated right bundle branch block. PMID- 11394323 TI - Analysis of radioactive particles from the Kola Bay area. AB - Two types of radioactive particle were identified in marine sediment and lichen samples collected from the Kola Bay, NW Russia. The particles were identified by means of gamma-ray spectrometry and autoradiography, separated and subjected to various analysis techniques. Several complementary techniques are needed to characterise particle properties thoroughly. 137Cs was present in the sediment matrix in large (approximately 100 microns) greenish particles that were most probably pieces of paint. Although their element composition was heterogeneous, 137Cs was found to be evenly distributed. 60Co in the lichen matrix was present in small (approximately 1 micron) particles. No U or transuranium elements were detected in either type of particle. PMID- 11394325 TI - Laryngoceles and saccular cyst. AB - Laryngoceles and saccular cysts, while relatively rare, occur more often than the literature would suggest. In the past, most were diagnosed clinically in a symptomatic patient. Today, however, more patients suspected of head and neck cancer are being diagnosed with diagnostic radiographic procedures. Of 2,068 patients studied at M.D. Anderson Hospital and Tumor Institute for suspected laryngeal or hypopharyngeal cancer, 87 (4.2%) had laryngoceles. There are three types of laryngoceles currently recognized: internal, external, and mixed/combined. Saccular cysts are divided into two categories: anterior and lateral. Both may be congenital, or acquired, and may occur at any age. This article reviews current evidence regarding classification, anatomy, clinical symptoms, etiology, pathogenesis, diagnostic techniques, and treatment of these two disorders. PMID- 11394326 TI - Radiology case of the month. Facial mass in a patient with end stage renal disease. Brown tumor of secondary hyperparathyroidism. PMID- 11394327 TI - Transmission of yellow fever by the culex mosquito: a look back. PMID- 11394328 TI - The Louisiana Cancer and Lung Trust Fund Board, 2000-2001. AB - The Louisiana Cancer and Lung Trust Fund Board, founded in 1980, continues to move forward in its mission in cancer control, cancer research, and tobacco control. This article summarizes the Board's activities of the past year, including revising the Cancer Control Strategic Plan and participating in a multi state development project funded by NCI to Bridge the Digital Divide. Listed at the end of this article is a summary of the Board's legislative mandate and its current membership. PMID- 11394329 TI - Racial variation in prostate specific antigen in a large cohort of men without prostate cancer. AB - Several studies have reported racial variation in serum prostate specific antigen (PSA) levels. Many of these studies, however, have included a significant number of men without a documented digital rectal examination (DRE) result or without prostate biopsies if abnormalities in PSA or DRE were detected. Thus, it is not clear that men with prostate cancer have been excluded in these analyses. In this report, data from 9,162 men (3,786 African-American men and 5,376 white men) are reviewed. All men had both serum PSA and DRE testing. Every man in this study had either a documented normal DRE and PSA (< 4 ng/mL) (3,422 African-American men and 4,795 white men) or a negative prostate biopsy (364 African-American men and 581 white men). Data were analyzed in age-matched decades. African-American men and white men had no difference in serum PSA levels between 30 and 39 years of age. At 40-49, 50-59, 60-69 and 70-79 years of age, African-American men had a statistically higher serum PSA level than white men. From these data, we conclude that racial variation in serum PSA is present in all decades above 40 years of age. Our data are unique in that this cohort included a substantial number of men between 30 and 39 years of age. In this group of young men, no racial differences in serum PSA were detected. These studies indicate, for the first time, that the onset of racial variation in PSA occurs after the fourth decade of life. PMID- 11394330 TI - Current practice and future trends in malignant melanoma. AB - The incidence of malignant melanoma is rising at a rapid pace. Since sun exposure is felt to be a causative factor for melanoma, it is necessary for Louisiana residents and physicians to have a heightened awareness about this tumor. Therefore, this article reviews the standard approach to prevention, diagnosis, staging, and treatment of melanoma. At the same time, new concepts in melanoma care are discussed, including sentinel lymph node mapping, immunotherapy, and melanoma vaccines. PMID- 11394331 TI - Breast cancer: incidence, mortality, and early detection in Louisiana, 1988-1997. AB - Breast cancer is the most commonly diagnosed cancer and the second leading cause of cancer death among Louisiana women. The incidence data from Louisiana Tumor Registry were used to calculate breast cancer incidence rates, which were compared with the combined rates from the National Cancer Institute's Surveillance, Epidemiology and End Results (SEER) program. Breast cancer mortality rates for Louisiana were compared with the US death rates from the National Center for Health Statistics (NCHS). Our data revealed that Louisiana women were not at a higher risk for developing breast cancer than women in the SEER areas, but that mortality rates in Louisiana were not correspondingly low. Although the percentage of cases diagnosed at an early stage (in situ and localized) increased in Louisiana from 1988 through 1997, the average in Louisiana was still below the level for the SEER areas (65.9% and 71.6%) in 1993 1997. The rates of in situ breast cancer significantly increased (on average 5.3% for whites per year and 7.1% for blacks), and localized breast cancer also significantly increased (2.6% for whites and 2.5% for blacks), while the incidence of distant stage breast cancer significantly decreased (3.4% for whites and 2.0% for blacks). Compared with white women, black women still were less likely to be diagnosed with early stage breast cancer in 1993-1997 (56.4% and 68.9%). Women residing in the parishes with high percentages of persons in poverty were less likely to be diagnosed with early stage of disease. PMID- 11394333 TI - Levers for change in medicine. PMID- 11394332 TI - Implementation of current Louisiana mammography legislation: a time for review. AB - Breast cancer mortality rates in the state of Louisiana are higher than national averages despite a lower than average incidence rate. Given the importance of early diagnosis in determining breast cancer outcomes, one potential explanation for relatively high mortality rates is an under-utilization of effective techniques for early detection. Comparison of mammography rates in Medicare recipients indicates that Louisiana mammography rates are among the lowest in the nation. To better assess barriers to mammography utilization, all certified mammography sites in Louisiana were contacted by telephone by a surveyor posing as a family member attempting to schedule a mammogram for a 50-year-old woman. Of the 151 sites in Louisiana, successful scheduling of a mammogram by phone was accomplished nine times. The vast majority of sites (94%) did not schedule a mammogram. Furthermore, 75.9% of the sites contacted provided no assistance to the consumer seeking a mammogram. Current Louisiana law requires that mammography be ordered by a licensed health care provider. Though compliance with current law is high, this law may provide a barrier to interested consumers attempting to schedule mammography. A revision of current law and policy may be indicated in an effort to increase mammography utilization rates. Such an effort may potentially help to decrease mortality from the second leading cause of cancer death in women. PMID- 11394334 TI - "The psychiatrist will be with you in a day or two". Unnecessary delays in assessing the mentally ill in police custody in Scotland. PMID- 11394335 TI - Policing and the mentally disordered. AB - I hope this article has given at least a flavour of the variety and complexity of challenges faced at the immediate operational level by police officers who have very little training in recognising the signs or symptoms of mental disorder and who very often have to deal with violent or irrational people who are quite simply criminal rather than ill. There is no question that in Dumfries Division police officers rely heavily upon the personal imput of on-call police doctors. Whilst at an operational level it is believed that incidents involvidng people with mental disorders are on the increase, it is particularly difficult to quantify exact extent of this perceived increase or indeed the impact which increasing abuse of alcohol and cotrolled drugs is having on the perception of mentally disordered offenders in the community. The vast majority of prisoners who are arrested and held within cells at Dumfries Police Office are suffering from some form of drug dependency and are in need of some form of assistance. Many display a variety of irrational behaviours when suffering the effects of drug withdrawal and increasingly it is difficult to keep any prisoner within the cells at Dumfries unless they have been examined by the on call police surgeon. Many are depressed, confused and increasingly providing at least verbal indicators that they are suicidal. It is more often the case that prisoners who are held within cells at Dumfries are in some form of constant observation whilst in our care. Given the increasing presence of mentally vulnerable people within our communities, it is vital that operational police officers play their part in responding appropriately to the needs of these people and the dangers they may pose to themselves or others. Dealing sensitively with increasing public concerns through appropriate community representative channels will also pose a considerable challenge. A challenge which can only be met through increasing awareness of the real situation and a commitment to enhance relationships across the caring and policing professions. The best chance of success in this area undoubtedly lies in the strength of these local relaitonships as progressed through honest and open inter-agency working. PMID- 11394336 TI - Police officers' knowledge of, and attitudes towards, mental illness in southwest Scotland. AB - Questionnaires were sent to all operational officers in a regional police service regarding their level of knowledge about and attitudes towards, mental illness. It was widely accepted by police officers that mental illness occurred commonly, can be effectively treated in the community and that the main risk of harm is to patients themselves. There was a good knowledge of relevant legislation, but most officers felt they did not have sufficient training in mental illness, and were keen for more. Improved liaison between sector psychiatrists and local police may be of value in the earlier identification and treatment of the mentally ill. PMID- 11394337 TI - Sudden unexpected infant deaths in Dundee, 1882-1891: overlying or SIDS? AB - Using a cohort study of all deaths in infants under 12 months in Dundee born between 1882-91 we set out to compare the aetiology of sudden unexpected infant deaths in Dundee at the end of the 19th Century with the aetiology of present day Sudden Infant Death Syndrome (SIDS). During 1882-1891, 361 infants died suddenly and unexpectedly and without obvious cause while in bed with their parents. The sex ratio of deaths was even (0.51 male) whereas the typical male fraction of SIDS today is 0.61. The mean age at death was almost two and one-half weeks younger in the Dundee cohort than for SIDS in modern Scotland. The infants in the Dundee cohort were discovered more frequently early in the morning than is typical. Their social class distribution was different in that no overlying cases were found in the higher classes whereas SIDS affects all classes. The overlying rate for illegitimate infants was lower than that reported for SIDS today. The epidemiological characteristics of the Dundee cohort and of those dying from present day SIDS differ considerably. The Dundee cohort apparently died from overlying rather than from SIDS as it is classified today. Present day advice that co-sleeping is safe should be given more cautiously until the safety of co sleeping is resolved. It might be prudent to inform parents that co-sleeping is a risk factor for SIDS and that it should therefore be avoided. PMID- 11394338 TI - Separating elective and emergency surgical care (the emergency team). AB - The purpose of this study was to evaluate the influence on general surgical activity following the separation of elective from emergency surgical care in one large teaching hospital. A prospective audit of elective and emergency general surgical activity between 1994 and 1999 inclusive was carried out. Elective and emergency surgical activity was separated in January 1996, with a dedicated 'Emergency Team' of one consultant for one week, two registrars, two senior house officers and four house officers for two weeks, in addition to a 20 bed acute admission ward and a 24 hour emergency theatre. The consultant cancelled the majority of his/her elective work during the on-call week. A prospective collection was made of all elective and emergency operations carried out between 1994 and 1999 using the Lothian Surgical Audit system. Out of hours operative activity was analysed retrospectively from data collected using the Operating Room Schedule of Surgery (ORSOS) and outpatient clinic and day case activity collected from the Hospital Administration System. Comparisons were made between years 1994/1995 and 1996/7/8/9. Emergency surgical admissions rose by 86% from 1973 patients in 1994 to 3675 in 1999. During the same period, elective in patient activity remained fairly steady, but there was an increase in day surgery from 469 to 2089 cases per annum. Despite the on-call consultant cancelling his/her outpatient clinics, overall outpatient activity also increased from 9911 to 12,335. However a proportion of this reflects the appointment of two new consultants in April 1998. Emergency operations increased from 941 in 1994 to 1351 in 1999, with a two-fold reduction in operations carried out between 0000 0800 hours from 16% in 1994 to 7.9% in 1999. A separate and dedicated 'Emergency Team' is an efficient method of managing acute general surgical admissions. It permits elective work to carry on uninterrupted, reduces the number of operations performed after midnight, and provides a better environment for teaching and training. This scenario might also be applicable to other medical specialties who have a large emergency commitment. PMID- 11394339 TI - Not the usual cause of superior vena cava obstruction. AB - Superior vena cava obstruction has a classical association with bronchial carcinoma. In the presence of cortisol excess, the diagnosis of small-cell carcinoma and ectopic ACTH production seems likely. We present a case where the clinical features did not accurately reflect the underlying pathology. PMID- 11394340 TI - Embolisation of renal arteriovenous malformation (AVM) in pregnancy. AB - A case of congenital renal arteriovenous malformation (AVM), presenting with profuse haematuria in the second trimester of pregnancy is reported. Selective embolisation with polyvinyl alcohol particles and absolute alcohol successfully ablated the lesion and a healthy infant was delivered at term five months later. Renal angiography at three years showed no evidence of the lesion. PMID- 11394342 TI - Tamoxifen therapy in steroid resistant Reidel's thyroiditis. AB - Riedel's thyroiditis is a rare chronic inflammatory disorder characterised by extensive fibrosis of the thyroid gland and sometimes the surrounding tissues. We report a case of Riedel's Thyroiditis in a middle aged female presenting with goitre, stridor and dyspnoea. She initially responded to corticosteroid treatment and subsequently to Tamoxifen. The rationale for these treatments are discussed. PMID- 11394341 TI - Rediscovering the joy of food: the need for long-term review of swallowing ability in stroke patients. AB - Stroke is commonly associated with dysphagia, which may necessitate the use of enteral feeding best provided by a percutaneous endoscopic gastrostomy (PEG) tube. Resolution of dysphagia complicating stroke is observed in the days, weeks and months following the event. This may allow the resumption of a normal diet, and cessation of enteral feeding. However at the time dysphagia resolves many patients--with enteral feeding still in place--will have been discharged from facilities where expert speech and language therapy is available to assess recovery of swallow. We report three cases of patients with dysphagia following stroke who benefited from long term review of swallowing ability. We then discuss the benefits of such follow-up, and how it might best be achieved in the community. PMID- 11394343 TI - Fractures in medieval Scotland. AB - The prevalence of fractures in medieval Scotland is assessed, particular attention being given to excavations of cemeteries beside three Carmelite cemeteries, at Aberdeen, Perth and Linlithgow, and another one at Whithorn Abbey. In the friaries the prevalence of fractures was 7.6% and in Whithorn it was 5.0%. These figures are comparable with an estimated prevalence of 7.2% for individuals between 0 and 65 years in present day Scotland. Males were more at risk of fractures than females, but a small group from both genders had been struck on the head by weapons. A study from a rural cemetery in England indicates that both male and female peasants had a much higher risk of fractures than their urban counterparts. PMID- 11394344 TI - The 54th Annual Meeting and Symposium of the Society of General Physiologists: structures and mechanisms of channels and transport proteins. PMID- 11394345 TI - Of filariasis, mice and men. PMID- 11394346 TI - Parasitology nomenclature--a recommendation. PMID- 11394347 TI - Seeking new targets for antiparasitic agents. PMID- 11394348 TI - Implications for neonatal HIV/AIDS and TB of sensitization in utero to helminths. PMID- 11394349 TI - DDT debate considering costs. PMID- 11394350 TI - Drugs and the immune system: the emerging era of immunopharmacology. PMID- 11394351 TI - Immunity, atherosclerosis and cardiovascular disease. PMID- 11394352 TI - Photo quiz: cerebral S. mansoni infection. PMID- 11394353 TI - The ten most commonly asked questions about transcatheter radiofrequency ablation. PMID- 11394354 TI - Surgery for chronic mitral valve regurgitation: determining the optimal time for intervention. PMID- 11394355 TI - How can we obtain maximal benefits from partial left ventriculectomy? PMID- 11394356 TI - TAR and the Y-graft: the golden myth, the tragic reality. PMID- 11394357 TI - Monitoring of palmer skin temperature in thoracoscopic sympathectomy. PMID- 11394358 TI - Stepping outside of the 'comfort zone' to communicate effectively. PMID- 11394359 TI - Why doesn't anyone ever listen to me? PMID- 11394360 TI - Information for patient. An overview of asthma treatment. PMID- 11394361 TI - Cardiovascular effects caused by rapid administration of gadoversetamide injection in anesthetized dogs. AB - RATIONALE AND OBJECTIVES: This study assessed the cardiovascular effects of gadoversetamide and other gadolinium chelates administered at high rates of injection. METHODS: Anesthetized beagles were instrumented to record the electrocardiogram and to measure arterial blood pressure. In part 1, each animal was injected with gadoversetamide at rates of 1.0, 3.0, and 10 mL/s. In part 2, each animal was injected with gadoversetamide, gadopentetate dimeglumine, gadodiamide, and gadoteridol at a dose of 0.6 mmol/kg delivered at a rate of 3.0 mL/s. RESULTS: Intravenous administration of gadoversetamide caused transient decreases in both heart rate and blood pressure. The rate of injection did not affect the magnitude of the heart rate or blood pressure changes. Administration of gadoversetamide, gadopentetate dimeglumine, and gadodiamide elicited equivalent changes in cardiovascular function. Injection of gadoteridol caused a similar degree of hypotension, but the changes lasted longer. CONCLUSIONS: Rapid administration of gadoversetamide caused no potentiation in cardiovascular changes. Our data support the initiation of a clinical trial to demonstrate the safety of rapidly administering gadoversetamide with the use of a power injector. PMID- 11394362 TI - Nonphysician groups push to expand scope of practice. PMID- 11394363 TI - Rejuvenation recipes. PMID- 11394364 TI - In the company of killers. PMID- 11394365 TI - Bibliography. Current world literature. Clinical therapeutics. PMID- 11394366 TI - Bibliography. Current world literature. Rheumatoid arthritis. PMID- 11394367 TI - Bibliography. Current world literature. Crystal deposition diseases. PMID- 11394368 TI - Bibliography. Current world literature. PMID- 11394369 TI - Value of the Doppler index of myocardial performance in the early phase of acute myocardial infarction. PMID- 11394370 TI - The influence of simvastatin on lipase and cholesterol esterase activity in the serum of men with coronary heart disease. AB - Several studies have demonstrated that any beneficial effects of 3-hydroxy-3 methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors (statins), of which simvastatin (Merck Sharp & Dohme) is an example, on coronary events are linked to their hypocholesterolemic properties. The in vivo effects of simvastatin treatment on lipase (GEH = glycerol ester hydrolase) and cholesterol esterase (CEase) activity in the serum of men with coronary heart disease (CHD) were examined. GEH and CEase activity in the serum of men with CHD, before simvastatin treatment, was lower than in the control subjects. In our study we have provided evidence that simvastatin increases GEH activity in a time-dependent manner, but has no effect on CEase activity. This suggests that simvastatin can directly affect acylglycerol metabolism by an increase in GEH activity and may therefore be suitable for the treatment of combined lipoprotein disorders characterized by elevation of triacylglycerols. PMID- 11394371 TI - Detection of the mandibular canal via shaded surface display and multiplanar reconstruction of CT data. AB - The purpose of this study was to evaluate the sensitivity of conventional two dimensional (2D) multisection images (multiplanar rendering, MPR) and registered three-dimensional (3D) shaded surface images (shaded surface display, SSD) of standard axial computed tomography (CT) data for detecting the mandibular canal (MC) in the lower jaw of 136 patients. The patients, who had different indications for mandibular CT, were examined using standard axial CT scanning. Two post-processing programs were used for 3D visualization of the data sets. The cross-sectional rendered images and the shaded surface 3D images were graded for detection of the MC, the presence of artefacts, overall quality and clinical relevance. A 3D display of the MC was achieved using the MPR technique in 100% with high image quality. The surface rendered display depicted the MC in 80%. Artefacts markedly degraded the 3D displays obtained using the surface rendering technique; thus, SSD is an inappropriate technique for imaging the entire MC without manual segmentation. MPR-CT improves the sensitivity of CT imaging in the detection of the MC with very little time needed for post-processing compared with the SSD method. This post-processing modality should, therefore, be considered for serial studies of patients undergoing dental CT. The sensitivity of the MPR method is even superior to the standard axial CT slices. PMID- 11394372 TI - Evaluation of biomechanical properties of Expanded-Polytetrafluoroethylene Soft Tissue Patch after dorsal implantation in the rat to mimic TMJ lateral reconstruction. AB - Clinically, Gore-Tex Expanded-Polytetrafluoroethylene (E-PTFE) has been used to reconstruct the lateral temporomandibular joint (TMJ) ligament. The purpose of this study was to assess changes in the biomechanical properties of implanted E PTFE over time with respect to tissue infiltration. Ninety-six specimens of implants were divided into four groups. Group A was the experimental group. Thirty-six autoclave-sterilized specimens were subcutaneously implanted into the backs of 36 rats. The rats were randomly sacrificed at 2 (n = 12), 7 (n = 12) and 12 (n = 12) weeks. The implants were tested for mechanical properties including maximal stress, strain and Young's modulus of elasticity (E) using the servo hydraulic material testing system (MTS). Group B was the in vitro control group. Thirty-six specimens were placed in tissue culture media at 37 degrees C for a time period equivalent to the experimental group to simulate the effect of a moist, warm environment on biomechanical properties. Group C was the temperature and pressure control group. Twelve specimens were autoclave-sterilized to determine the changes of tensile strength under high temperature and pressure. Control group D (no treatment) was tested to determine the initial tensile strength. The results showed significantly larger maximal stress as well as an increase in E and smaller maximal strain in experimental group A than in control groups B, C and D. There was no significant difference among control groups B, C and D. Histological examination of implants at 12 weeks demonstrated that 0.2-0.3 mm of 1-mm thick implants were occupied by connective tissue from each side. It may be concluded that E-PTFE implants become stronger and less flexible after implantation in vivo. PMID- 11394373 TI - Position of teeth on the edentulous atrophic maxilla. AB - For prosthetic treatment of strongly atrophic alveolar wall, some biometric methods have been developed. The measurements taken from plaster cast models of 230 edentulous, average 29.9 years, and 125 dentulous post-menopausal women were correlated. In the edentulous maxilla the sagittal position of canine teeth can be determined by the oral edge of incisive papilla. The transverse position of canine teeth was on the outer edge of the alveolar wall because of the extensive loss of buccal alveolar bone. On the incisor area the facial surfaces of the central incisors were determined by the oral edge of incisive papilla and the distance was about twice the length of the papilla. The sagittal position of the first premolars was one-third and the first molars two-thirds the length of the palate from the plane of the labial edge of incisive papilla. The transverse position of the premolars and molars was determined by the scar-line, which is a cord-like elevation or track on the alveolar mucosa after extractions of the teeth. According to the comparative method, the position of the scar-line differed from the lingual gingival margin line and was situated about half breadth of the tooth in a buccal direction from it. The transverse position of premolar and molar in the edentulous maxilla is about the middle of the scar-line in a facio-buccal direction. In the setting of the artificial teeth, the facial surfaces of these teeth should be on average 5.0-6.0 mm sideways from the scar line, whilst the total bilateral breadth of the alveolar wall in the sulcus area was on average 1.0-2.0 mm larger. PMID- 11394375 TI - Prevalence of temporomandibular dysfunction in Turkish children with mixed and permanent dentition. AB - This study was carried out to determine the prevalence of temporomandibular joint (TMJ) dysfunction in mixed and permanent dentition and to evaluate the sex distribution in Turkish children. One hundred and eighty-two children with mixed dentition and 212 with permanent dentition were selected for the study, which used a questionnaire and clinical examination. Children with one or more signs (TMJ sounds, TMJ tenderness, muscle tenderness, restricted mouth opening) and/or symptoms (TMJ pain during mastication and mouth opening, restriction of the jaw opening and TMJ sounds) met the criterion of TMJ dysfunction. The total prevalence of signs and symptoms of TMJ dysfunction in the studied population was 68% (68% in girls and 68% in boys) in mixed dentition and 58% (61% in girls and 56% in boys) in permanent dentition. The Z-test and analysis of variance (ANOVA) were used for statistical analysis of the difference between the results. The total prevalence of signs and symptoms of TMJ dysfunction in mixed dentition was found to be higher than in permanent dentition (P < 0.05). No statistically significant difference was found in the total prevalence of TMJ dysfunction between girls and boys. PMID- 11394374 TI - Mechanical properties of human articular disk and its influence on TMJ loading studied with the finite element method. AB - The present study was designed to investigate the elastic modulus of human temporomandibular joint (TMJ) disk under tension and its influences on TMJ loading. Seven human TMJ disks served as specimens. Continuous tensile stress was applied to each specimen, and the elastic moduli of human TMJ disks were calculated at 2% strain. Furthermore, using a three-dimensional finite element model of the mandible including the TMJ, changes in the TMJ stresses during clenching were evaluated in association of varying elastic moduli of the articular disk determined by the tensile tests. The elastic moduli at 2% strain varied from 27.1 to 65.2 MPa with a mean of 47.1 MPa. A significant correlation was found between the elastic moduli and age (P < 0.01). On the surface of condyle, compressive stress in the anterior area and tensile stress in the posterior area increased when the elastic moduli of the TMJ disk was varied from 25 to 65 MPa. In the TMJ disk, shear stresses in all the areas became larger with greater stiffness. In conclusion, it is shown that the elastic modulus of human TMJ disk is increased with age and that higher stiffness of the disk exerts substantial influences on mechanical loading for the TMJ structures. PMID- 11394376 TI - Application of principal component analysis to the elucidation of perioral soft tissue movements during mastication. AB - We considered that elucidation of the movements of perioral soft tissue during mastication would be useful in evaluating masticatory movements. However, the evaluation of these movements is difficult because multivariable analysis is needed. In this study, we considered whether principal component analysis (PCA), a form of the multivariate analysis, can reduce the number of degrees of freedom (d.f.) of perioral skin movements (i.e. the time-series data with 45 d.f.). The subjects were 15 healthy persons with complete natural dentition. The chosen experimental food for this study was sufficiently softened chewing gum. Over 95% of the perioral soft tissue movements of healthy subjects with complete natural dentition during mastication could be expressed by PCA using the first three principal components (PCs). Therefore, perioral soft tissue movements in these subjects during mastication were considered to be spatially smooth. Moreover, time analysis of these movements was made possible by the application of proportion diagrams. The results of this study showed that the spatiostructural and temporal analyses of the movements of perioral soft tissue during mastication made possible by the application of PCA. PMID- 11394377 TI - Dextromethorphan is an effective cough suppressant. PMID- 11394378 TI - 16q subtelomeric deletion in proband with congenital malformations and mental retardation. AB - We present a female child with mild mental retardation and congenital malformations. After fluorescence in situ hybridization (FISH) we found only abnormal karyotype in all cells. We used rapid FISH and original DNA probes- PAC62.10.1 and PAC20.19.N, specific for segments of chromosome 16q24. Karyotype of proband 46,XX.ish del(16)(q24.2:) (PAC20.19.N,PAC62.10.1-). Parent karyotypes are normal. This case may suggest the presence of clinical picture 16q- with defined clinical polymorphism at small telomeric loss, and also its necessary of the use of molecular-cytogenetic techniques in genetic departments. PMID- 11394379 TI - Flat and depressed colonic neoplasms: a prospective study of 1000 colonoscopies in the United Kingdom. PMID- 11394380 TI - Value of endoscopic ultrasound guided fine needle aspiration biopsy in the diagnosis of solid pancreatic masses. PMID- 11394381 TI - A new variable stiffness colonoscope makes colonoscopy easier: a randomized controlled trial. PMID- 11394382 TI - Surgery of the penis: reconstruction and prostheses. AB - Implantation of penile prostheses was a very popular topic in the published literature on reconstructive urological surgery in the year 2000. Monocomponent, multicomponent, semirigid and inflatable prosthesis techniques were investigated. The best results were obtained with multicomponent inflatable prostheses. For penile reconstruction, various techniques were described. All authors used myocutaneous sensitive forearm flaps. Good results were reported. PMID- 11394383 TI - [The 74th general meeting of the Japanese Leprosy Association. Tottori, Japan. May 10-12, 2001. Abstracts]. PMID- 11394384 TI - Photo quiz. Diagnosis: apical fibrocavitary disease associated with ankylosing spondylitis. PMID- 11394385 TI - Survival for colorectal cancer. PMID- 11394386 TI - Helicobacter pyroli, Lewis antigens, and inflammation. PMID- 11394387 TI - Relation of H pyroli to gastric mucins and gastric surface mucous gel layer. PMID- 11394388 TI - Helicobacter pyroli infection and acid secretion in patients with duodenal ulcer in Japan. PMID- 11394389 TI - Angiography. PMID- 11394390 TI - Frequent association between trisomy 15 and t(8;21)(q22;q22) in acute myeloid leukemia. PMID- 11394391 TI - Predictable multiple site root coverage using an acellular dermal matrix allograft. AB - BACKGROUND: The primary aim of this randomized, controlled, blinded clinical investigation was to determine if orientation of an acellular dermal matrix (ADM) allograft, basement membrane side against the tooth or connective tissue side against the tooth, affected the percent root coverage. Additional aims were to: 1) compare results of this study with results obtained from other root coverage studies; 2) determine if multiple additional sites could be successfully covered with the same surgery; 3) determine the effect of the procedure on keratinized tissue; and 4) evaluate the amount of creeping attachment obtained. METHODS: Ten patients with 2 Miller Class I or II buccal recession defects > or =3 mm were treated with a coronally positioned flap plus ADM and followed for 12 months. Test sites received ADM with the basement membrane side against the root (AB), while the control sites received the connective tissue side against the root (AC). Multiple additional recession sites were treated with the same flap procedure. RESULTS: Mean baseline recession for the AB sites was 4.2 mm and for the AC sites, 3.7 mm. Mean root coverage of 95% was obtained for both AB and AC sites. Sixty-eight additional Class I or II AB and AC sites obtained about 93% root coverage. The mean increase in keratinized tissue for both treatments was 0.80 mm. No additional root coverage was gained due to creeping attachment between 2 and 12 months. CONCLUSIONS: Treatment with ADM was an effective and predictable procedure for root coverage. The orientation of the material did not affect the treatment outcome for any of the parameters tested. PMID- 11394392 TI - Combination use of bovine porous bone mineral, enamel matrix proteins, and a bioabsorbable membrane in intrabony periodontal defects in humans. AB - BACKGROUND: The purpose of this study was to evaluate the effectiveness of a combination of enamel matrix proteins (EMP), bovine porous bone mineral (BPBM), and a bioabsorbable membrane for guided tissue regeneration (GTR) as regenerative therapy for intrabony defects in humans and compare it to an open flap debridement (OFD) technique. METHODS: Using a split-mouth design, 18 pairs of intrabony defects were treated and surgically reentered 6 months after the initial surgery. Experimental sites were treated with EMP, grafted with BPBM, and received a collagen/polylactic acid membrane for GTR. Control sites were treated with OFD. The primary outcomes evaluated in the study included probing depth resolution, clinical attachment gain, and bony defect fill. RESULTS: Preoperative probing depths, attachment levels, and intraoperative bone measurements were similar for the experimental and control groups. Postsurgical measurements taken at 6 months revealed a significantly greater reduction in probing depth in the experimental group (4.95+/-1.52 mm on buccal sites and 4.74+/-1.47 mm on lingual sites) when compared to the control group (2.83+/-0.83 mm on buccal sites and 2.90+/-0.91 mm on lingual sites). The experimental sites also presented with significantly more attachment gain (3.89+/-1.16 mm on buccal sites and 3.78+/ 1.14 mm on lingual sites) than the control sites (1.52+/-0.83 mm on buccal sites and 1.48+/-0.78 mm on lingual sites). Surgical reentry of the treated defects revealed a significantly greater amount of defect fill in favor of the experimental group (4.76+/-1.36 mm on buccal sites and 4.81+/-1.37 mm on lingual sites) as compared to the control group (1.78+/-0.92 mm on buccal sites and 1.67+/-0.90 mm on lingual sites). CONCLUSIONS: The results of this study indicate that a combination technique including BPBM, EMP, and GTR results in better clinical resolution of intrabony defects than treatment with OFD. Differences observed were both statistically and clinically significant. The exact role of each of the 3 technique components in achieving the clinical improvement observed in this study remains to be determined. PMID- 11394393 TI - Expression of cytokines and inducible nitric oxide synthase in inflamed gingival tissue. AB - BACKGROUND: Periodontopathic bacteria induce inflammation of periodontal tissues. The cytokines and nitric oxide released in periodontal lesions have been reported to play a protective role in bacterial infection and to relate to the process of inflammation. To clarify the relationship between colonization of periodontopathic bacteria and cytokines, we evaluated profiles of inflammatory cytokines, chemokine, anti-inflammatory cytokines, and inducible nitric oxide synthase (iNOS) and colonization by Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans, which are major pathogens of periodontitis. METHODS: mRNA expression of cytokines and iNOS in inflamed and healthy gingival tissue was determined using reverse transcription-polymerase chain reaction (RT-PCR), and the relationship between their profiles and the detection of specific bacteria was analyzed. RESULTS: The relative expression of interleukin (IL)-6 and iNOS mRNAs in periodontal lesions was significantly higher than those in healthy individuals. IL-6 mRNA expression was also significantly higher at bleeding on probing (BOP)-positive sites than at BOP-negative sites. The expressions of IL 1alpha and IL-8 increased, but IL-10 expression decreased at sites where A. actinomycetemcomitans was detected. We found no correlation between the expression of cytokine and iNOS mRNA and infection by P. gingivalis. CONCLUSIONS: The expression of IL-6 may reflect inflammation in gingival tissue, and iNOS may be involved in the inflammatory process in periodontitis. The presence of A. actinomycetemcomitans or P. gingivalis might relate to the different cytokine profiles of IL-1alpha, IL-8, and IL-10. PMID- 11394394 TI - Human fibroblasts ubiquitously express glutamic acid decarboxylase 65 (GAD 65): possible effects of connective tissue inflammation on GAD antibody titer. AB - BACKGROUND: Type 1 diabetes is caused by a destruction of pancreatic beta cells due to autoimmunity. Autoantibody against glutamic acid decarboxylase (GAD) 65 expressed in pancreatic beta cells is widely used as a predictive marker for pancreatic destruction. In this study, we hypothesized that if certain cells in periodontal tissues could express GAD, then it may influence GAD antibody titer. METHODS: We used: 1) reverse transcription-polymerase chain reaction (PCR) analysis to detect GAD 65 mRNA in various cells; 2) nucleotide sequencing analysis to confirm that amplified PCR product is the gene encoding GAD; and 3) Western blotting to determine the expression of GAD 65 protein in human gingival fibroblasts. Immunohistochemical staining of GAD 65 protein in normal and inflamed gingiva was performed to examine the potential influence of periodontal inflammation on GAD 65 expression. GAD antibody titer in sera of periodontal patients as well as healthy subjects was measured to determine if periodontal patients could develop autoantibody against GAD 65. RESULTS: Cultured human gingival, periodontal, and dermal fibroblasts and mesangial cells expressed GAD mRNA. Nucleotide sequencing analyses confirmed the amplified PCR product as GAD 65. Western immunoblotting analyses and immunohistochemical staining revealed that the GAD 65 protein was expressed in vitro and in vivo. The expression of GAD 65 in inflamed tissue was higher than that in normal tissues. Two of 62 periodontal patients without diabetes showed an increased antibody titer against GAD 65, while none of the systemically healthy subjects showed an increased antibody titer against this antigen. CONCLUSIONS: We concluded that periodontal inflammation may result in higher levels of GAD and influence GAD antibody titer, and, hence, affect diabetic diagnosis based upon GAD antibody production. PMID- 11394395 TI - Prevalence and risk of gingival enlargement in patients treated with nifedipine. AB - BACKGROUND: Gingival enlargement is a known side effect of nifedipine use. This study was conducted to determine the prevalence and risk factors for gingival enlargement in nifedipine-treated patients. METHODS: A cross-sectional study was conducted in a primary care center. Data from 65 patients taking nifedipine were compared with 147 controls who had never received the drug. All patients were examined for the presence of gingival enlargement using 2 different indices: vertical gingival overgrowth index (GO) in 6 points around each tooth, and horizontal MB index in the interdental area. Gingival index, plaque index, and probing depth were also evaluated. RESULTS: The prevalence of gingival enlargement was significantly higher in nifedipine-treated cases than in controls (GO index, 33.8% versus 4.1%; MB index, 50.8% versus 7.5%, respectively). Higher gingival and plaque indices were observed in patients taking nifedipine. Among the possible risk factors, only the gingival index showed a significant association with gingival enlargement. The risk (odds ratio [OR]) of gingival enlargement associated with nifedipine therapy was 10.6 (3.8-29.1) for the GO index and 14.4 (6-34.6) for the MB index. Gingival index-adjusted ORs were 9.6 (3.3-28.1) and 9.7 (3.9-23.3), respectively. In the subset of high nifedipine exposure patients, the odds ratio for gingival enlargement increased to 17.4 (5.3 56.3) for the GO index and 23.6 (7.7-72.3) for the MB index. The concordance between GO and MB indices showed a kappa value of 0.689 in controls and 0.642 in patients treated with nifedipine. CONCLUSIONS: Patients taking nifedipine are at high risk for gingival enlargement, and gingivitis acts as a predisposing factor. PMID- 11394396 TI - Root trunk concavities as a risk factor for regenerative procedures of class II furcation lesions in dogs. AB - BACKGROUND: The purpose of this study was to evaluate the influence of root trunk concavities on guided tissue regeneration. METHODS: The second, third, and fourth mandibular bicuspids of 4 healthy mongrel dogs were used. Full flaps were elevated and furcation lesions (5 mm in height and 2 mm in depth) were surgically created. One mm deep concavities were prepared on the root trunks and part of the crown of all teeth. The second bicuspids remained as controls (C); the left third and fourth bicuspids received normal expanded polytetrafluoroethylene (ePTFE) membranes (NM) and the right bicuspids received modified ePTFE membranes (MM). The MM were made by removing the collar from an NM, cutting it into 2 mm segments, and suturing each of the segments to the collar of each MM. Normal and modified membranes were sutured to their corresponding teeth and the flaps sutured. After the healing period, the mandibles were removed and the teeth processed for histomorphometry. RESULTS: The results showed that the junctional epithelium measured a mean of 0.71 mm for C, 0.64 mm for NM, and 0.14 mm for MM. The differences between C and MM and between NM and MM were both statistically significant at the 5% level (Kruskal-Wallis test). Mean bone height measurements for the buccal surfaces from the furcation roofs were 2.79 mm for C, 2.60 mm for NM, and 1.06 mm for MM and for the mid-portion 1.10 mm for C, 1.23 mm for NM, and 0.30 mm for MM. Differences were statistically significant at the 1% level between C and MM and between NM and MM for the buccal measurements, and at the 5% level between NM and MM for the mid-portion measurements. CONCLUSIONS: Root trunk concavities are important risk factors for regenerative procedures. The collars of the membranes should be modified to improve results when concavities are present. PMID- 11394397 TI - Naturally occurring periodontal bone loss in the wild deer mouse, genus Peromyscus. AB - BACKGROUND: Historically, animal models for the study of periodontal diseases have incorporated surgically created defects, plaque retentive ligatures, as well as soft and high-sucrose diets which may not accurately reflect progression of the natural disease. Spontaneous periodontal disease is seen in a few animal species, but these are often expensive to maintain and are unsuitable for manipulation using advanced molecular biology techniques. Mice are inexpensive, easy to maintain, and are routinely used for transgenic experiments and are therefore an optimal animal for research purposes. However, it is commonly accepted that mice do not spontaneously develop periodontal disease. The purpose of this study was to determine if a mouse population that exhibits periodontal breakdown in the wild could be found, allowing for genetic manipulation of naturally occurring periodontal disease. METHODS: We examined over 2,500 dry skulls of several Peromyscus species from various locations and habitats on the west coast of North America for periodontal bone loss in the molars, using furcation involvement as an indicator of disease severity. Alveolar bone loss was classified as Grade I) horizontal component of bone loss in the furcations; II) through-and-through furcations; and III) through-and-through furcations with alveolar bone loss into the apical third of the root. RESULTS: The proportions of individual mice experiencing bone loss were 3.8% for Class I-III involvement, 1.3% for Class II-III involvement, and 0.5% for Class III alone. Three subspecies of P. keeni and one subspecies of P. maniculatus had periodontal disease prevalences in 7% to 13.5% of their samples. Mice from isolated islands had 1.8- to 4.7-fold higher disease prevalence than those located on the mainland, with even greater prevalence on small islands. No statistically significant differences between genders were found. CONCLUSIONS: It appears that periodontal disease is far more common in this mouse genus than previously believed. Some of the subspecies demonstrated severe periodontal disease at a prevalence comparable to that found in humans. PMID- 11394398 TI - Adhesion of Porphyromonas gingivalis strains to cultured epithelial cells from patients with a history of chronic adult periodontitis or from patients less susceptible to periodontitis. AB - BACKGROUND: The present study aimed to explain the interindividual variation in periodontitis susceptibility by differences in the initial adhesion rate of Porphyromonas gingivalis to the pocket epithelium of these individuals, and/or by inter-P. gingivalis strain differences in association capacity (adhesion and internalization). METHODS: Adhesion assays were performed on epithelial monolayers (cultured in vitro from pocket epithelium belonging to patients who were less or more susceptible to chronic adult periodontitis) using 11 genetically different clinical strains of P. gingivalis. RESULTS: Both the disease category (less susceptible versus susceptible) and the interstrain variation were found to have a significant effect (both P <0.05) on the initial bacterial association. The chronic adult periodontitis group showed significantly more association of P. gingivalis when compared to less susceptible patients (4.2 x 10(6) versus 3.5 x 10(6)). Also, the interstrain variation was significant, with strains Pg 4 and 5 representing the least and best associating bacteria (1.8 x 10(6) colony forming units for Pg 4, 9 x 10(6) for Pg 5). CONCLUSIONS: These results indicate that periodontitis susceptibility is influenced by both the interindividual differences in pocket epithelium (allowing more adhesion of P. gingivalis) or by the strain type by which the patient is infected (intra-species differences in adhesion capacity). PMID- 11394399 TI - Soluble CD14 levels in gingival crevicular fluid of subjects with untreated adult periodontitis. AB - BACKGROUND: This study determined soluble CD14 (sCD14) levels in gingival crevicular fluid (GCF) and their potential relationship to periodontal conditions in adult periodontitis. METHODS: GCF was collected from 15 patients with untreated adult periodontitis. sCD14 levels were determined by ELISA and presented as total amount (ng/site) and concentration (microg/ml). The periodontal examination consisted of plaque index (PI), bleeding index (BI), probing depth (PD), and clinical attachment level (CAL). PD and CAL were measured with an electronic probe. RESULTS: sCD14 was detected in all 15 subjects and was found in 59% (62/105) of the sampled sites. The percentage of sites with sCD14 varied greatly, ranging from 14% to 100%. The mean total amount of sCD14 was 1.71+/-0.40, range 0.03 to 5.41 ng/site; the concentration of sCD14 was 14.04+/ 4.15, range 0.16 to 51.74 microg/ml. No significant difference in clinical data was found between the sites with and without detectable levels of sCD14. However, on the basis of the individual profile of sCD14 levels, i.e., those individuals with >50% of the sites containing sCD14 and mean levels of sCD14 >5.0 microg/ml, the 15 subjects were divided into a high sCD14 group (9 subjects) and a low sCD14 group (6 subjects). Compared to the high group, the low group showed greater mean PD and a higher percentage of sites with PD > or = 5.0 mm (P <0.05). Consistent with this, sCD14 concentrations showed a negative correlation with PD (r(s) = 0.636, P = 0.0174). CONCLUSIONS: The present study shows that sCD14 levels in GCF varied greatly among subjects with untreated adult periodontitis. Individuals with higher levels of sCD14 in GCF and more sites containing sCD14 had fewer deep pockets. The negative correlation between GCF sCD14 levels and probing depth implies a crucial role of sCD14 in bacterially induced periodontal destruction. The relationship between GCF sCD14 levels and probing depth warrants further investigations. PMID- 11394400 TI - Protease-active extracellular protein preparations from Porphyromonas gingivalis W83 induce N-cadherin proteolysis, loss of cell adhesion, and apoptosis in human epithelial cells. AB - BACKGROUND: The protease-induced cytotoxicity of P. gingivalis may partly result from alteration of the extracellular matrix and/or surface receptors that mediate interaction between the host cells and their matrix. While P. gingivalis-induced degradation of E-cadherin has been documented, there is no information on the effects of P. gingivalis proteases on other members of this family of cell adhesion proteins. METHODS: Human epithelial KB cells were exposed to protease active extracellular protein preparations from isogenic mutants of P. gingivalis. Quantification of apoptosis was performed by visualization of nuclei stained with 4,6'-diamidino-2-phenylindole. Alteration of cell adhesion proteins was examined by immunoblotting of cell lysates using monoclonal antibodies to those proteins. RESULTS: Treated cells exhibited loss of cell adhesion properties with apoptotic cell death subsequently observed. These effects correlated with the different levels of cysteine-dependent proteolytic activities of the isogenic mutants tested. Cleavage of N-cadherin was observed in immunoblots of lysates from detached cells. There was a direct correlation between the kinetics of N-cadherin cleavage and loss of cell adhesion properties. Loss of cell adhesion, as well as N-cadherin cleavage, could be inhibited by preincubation of P. gingivalis protease active extracellular protein preparations with the cysteine protease inhibitor TLCK. In control experiments, the cleavage of N-cadherin was detected after treatment of KB cells with trypsin but not after cell dissociation by a non enzymatic method. CONCLUSIONS: These results suggest that extracellular proteases from P. gingivalis can induce degradation of N-cadherin, which could have implications for the pathogenicity of this bacterium. PMID- 11394401 TI - Hyaluronan supports recombinant human bone morphogenetic protein-2 induced bone reconstruction of advanced alveolar ridge defects in dogs. A pilot study. AB - BACKGROUND: Prosthetic-driven implant dentistry requires predictable procedures for alveolar ridge augmentation. The objective of this pilot study was to evaluate bone regeneration in mandibular, full-thickness, alveolar ridge, saddle type defects following surgical implantation of recombinant human bone morphogenetic protein-2 (rhBMP-2) in a novel hyaluronan (HY) sponge carrier. This sponge was fabricated from auto-crosslinked HY. METHODS: Alveolar ridge defects (approximately 15 x 10 x 10 mm), 2 per jaw quadrant, were surgically prepared in each of 3 young adult American fox hounds. Four defects were immediately implanted with rhBMP-2/HY. Three defects were implanted with rhBMP-2 in an absorbable collagen sponge (ACS) carrier (positive control). The rhBMP-2 solution (1.5 ml at 0.2 mg/ml) was soak-loaded onto the HY and ACS sponges. Three defects were implanted with HY sponges soak-loaded with buffer without rhBMP-2 (negative control), while 2 defects served as surgical controls. The animals were euthanized at 12 weeks postsurgery for histometric analysis. RESULTS: Clinically, alveolar ridge defects receiving rhBMP-2/ACS exhibited a slight supracrestal expansion, while defects receiving rhBMP-2/HY were filled to contour. In contrast, the HY and surgical controls exhibited ridge collapse. rhBMP-2/HY treated defects exhibited a dense bone quality without radiolucent regions observed in defects treated with rhBMP-2/ACS. The histometric analysis showed 100% bone fill for the rhBMP-2/ACS defects and 94%, 58%, and 65% bone fill for the rhBMP-2/HY, HY, and surgical control defects, respectively. CONCLUSIONS: The conclusions are based on data from 2 of 3 animals in the study. In one animal, no response to rhBMP-2 was observed with either carrier, and the animal may have been a non-responder of unknown nature. With this limitation, the observations herein suggest that: 1) HY supports significant bone induction by rhBMP-2; 2) the rhBMP-2-induced bone assumes qualities of the immediate resident bone; 3) HY alone exhibits no apparent osteoconductive potential; and 4) HY appears to resorb within a 12-week healing interval in the absence or presence of rhBMP-2. Thus, HY appears to be a suitable candidate carrier for rhBMP-2. PMID- 11394402 TI - Effects of cyclosporin A on dental alveolar bone: a histomorphometric study in rats. AB - BACKGROUND: We have previously reported cyclosporin A (CA)-induced osteopenia around the dental alveoli of the mandibular incisors of rats. The drug-induced tooth displacement and the regional anatomical complexity around the mandibular incisors might complicate the local effects of CA. Therefore, the purpose of this study was to evaluate the dental alveolar bone histomorphology around maxillary secondary molars in CA-treated rats and to further elucidate the effects of CA on the dental alveolus. METHODS: Twenty Sprague-Dawley rats were assigned to a CA and a control group. Animals in the CA group received CA (15 mg/kg) daily and the control rats received only mineral oil. At the end of weeks 2 and 4, five animals in each group were sacrificed. Dental alveoli around the maxillary second molar region were frontally sectioned and stained with toluidine blue by undecalcified histological processing. Ten serial tissue sections, 80 microm apart, were selected for histometric evaluation. Bone volume, bone-specific surface, and osteoid formation were measured at buccal, apical, and palatal locations in dental alveolus. RESULTS: Overall bone mass in dental alveolus decreased more in the CA group than in the control group at both observation intervals. All histometric measurements, except the bone-specific surface, were significantly affected by the alveolar location (palatal, apical, and buccal) and CA therapy (P= 0.004 and <0.001, 0.001 and <0.001, 0.004 and <0.001 for drug therapy and location of the dental alveolus in bone volume, marrow volume, and the ratio of bone surface to volume, respectively). Decreased bone volume, but increased marrow volume, were noted in the CA group compared to the control group. Although the alveolar bone surface area did not differ between the CA group and the control group, greater alveolar surface-to-volume ratio was noted in the CA group. For osteoid, more decreased volume, seam width, and fractional formation surface were observed in the CA group compared to the control group (P <0.001, <0.001, and = 0.046 in osteoid volume, seam width volume, and formation surface, respectively). CONCLUSIONS: Because the bone mass and the osteoid formation in the dental alveolus around the maxillary molar region showed a decrease after CA exposure, we conclude that this drug has inhibitory effects on the dental alveoli. PMID- 11394403 TI - Smoking affects the subgingival microflora in periodontitis. AB - BACKGROUND: Tobacco smoking has been identified as one major risk factor for destructive periodontal disease. Scaling and root planing have been shown to be less effective in smokers with periodontitis. The aim of the present study was to compare the subgingival microbial flora of treated and untreated smokers and non smokers. METHODS: Four independent adult patient groups with periodontitis were included in this investigation: 88 untreated smokers (U-S); 90 untreated non smokers (U-NS); 119 treated non-smokers (T-NS); and 171 treated smokers (T-S). Clinical variables included cumulative plaque index (CPI), probing depth (PD), clinical attachment level (CAL), cumulative bleeding index (CBI), and cumulative suppuration index (CSI). Paper point samples from the deepest bleeding pocket in each quadrant of the dentition were analyzed for the presence and levels of 6 periodontal bacterial pathogens using anaerobic culture techniques. RESULTS: U-S showed a higher mean cumulative plaque index than U-NS (3.5 versus 2.7). Mean PD and mean CAL were higher in the T-S in comparison to the T-NS group (7.0 versus 6.6 mm and 5.6 versus 4.7 mm, respectively). Microbiological characteristics of U S were a higher prevalence of Prevotella intermedia/nigrescens and higher mean levels of Peptostreptococcus micros (Pm) and Fusobacterium nucleatum (Fn). T-S patients were characterized by higher prevalence of Bacteroides forsythus (Bf), Pm, and Campylobacter rectus (Cr) and higher mean levels of Pm and Fn. The mean percentage of B. forsythus tended to be higher in the T-S group than in the T-NS group (6.9% versus 5.6%). The relative risk to be infected with Bf, Pm, and Cr was statistically higher in smokers (odds ratios: 1.9, 1.9, and 1.6, respectively). The chance to find > or =10% of Bf, Pm, and/or Fn was 3.3 higher in smokers when A. actinomycetemcomitans and P gingivalis were absent. Detection of > or =20% Pm/Fn in treated patients was strongly associated with smoking (odds ratio 13.8, P= 0.002). CONCLUSIONS: Smoking is a determining factor for the composition of the subgingival microflora in adult patients with periodontitis and may select for a specific cluster of periodontal pathogens, notably Bf, Pm, Fn, and Cr. On the basis of these observations, smoking, among other criteria, may be one parameter to use in deciding to treat refractory periodontitis in smokers with a systemic antibiotic therapy directed against smoking-associated periodontal bacteria. PMID- 11394404 TI - Gliding capacity of different dental flosses. AB - BACKGROUND: One of the major concerns with the use of dental floss is the passage of the proximal contact, which might be influenced by the material characteristics of the floss. The aim of this study was to compare the gliding capacities of different flosses with major differences in structure and experimental behavior in vivo. METHODS: In a clinical, single-blind, crossover study of 27 subjects, the forces necessary for passing all 14 proximal contacts between the first premolars were measured using 2 polytetrafluoroethylene (PTFE) based flosses, 2 nylon-based flosses, and 1 nylon-based tape. The flosses were fixed in a special holder with strain gauges, which allowed registration of the maximum forces at insertion into and removal from the interdental area. RESULTS: In most contacts, the PTFE flosses needed less force compared to the nylon flosses. The range between the flosses was higher in the mandible compared to the maxilla and increased from the mesially located contacts to the more distally located contacts. The nylon tape required higher forces at insertion for all maxillary contacts. All other flosses proved to be equivalent at these contacts. CONCLUSIONS: PTFE flosses are superior to nylon flosses in terms of passing stronger proximal contacts in both directions. Due to gliding differences between different types of floss, the selection of a floss has the potential of compensating intra- and interindividual variation in contact strengths. PMID- 11394405 TI - The effect of enamel matrix protein derivative on follicle cells in vitro. AB - BACKGROUND: It is thought that during development of the periodontium, dental follicle cells, when appropriately triggered, have the ability to differentiate into periodontal ligament fibroblasts, cementoblasts, and osteoblasts. However, the exact mechanisms/factors responsible for initiating cell differentiation are not defined. The purpose of this in vitro study was to further characterize follicle cells and to determine the effects of an enamel matrix-derived protein (EMD) on these cells. METHODS: Murine follicle cells, transformed with simian virus 40 (SV 40) T antigen-containing virus (SVF cells), were used. SVF cells were cultured in Dulbecco's modified Eagle's medium (DMEM) plus 2% fetal bovine serum (FBS) or 2% FBS plus EMD (100 microg/ml), with and without ascorbic acid (50 microg/ml). For proliferation assays, cells were plated at 500 cells/cm2 in 24-well plates and counted on days 3, 4, and 5. For Northern analysis, total RNA was isolated on days 8, 12, and 18. Induction of mineral nodules by SVF cells was determined by von Kossa staining. RESULTS: EMD had a significant proliferative effect on SVF cells, when compared with 2% FBS control. Based on investigations in situ, follicle cells at the time point used here do not express key mineral associated markers, e.g., osteocalcin (OCN) or bone sialoprotein (BSP). Significantly, by day 12 in culture, Northern analysis indicated that the follicle cells expressed transcripts for BSP, OCN, and osteopontin (OPN). EMD increased OPN mRNA and decreased OCN mRNA expression. SVF cells were capable of inducing mineralization on day 18, but EMD blocked this activity. CONCLUSIONS: These results suggest the follicle cells have the capacity to act as cementoblasts or osteoblasts. Furthermore, EMD can regulate follicle cell activity, thus suggesting that epithelial-mesenchymal interactions may be important during development of periodontal tissues. PMID- 11394406 TI - Metastatic transitional cell carcinoma of the urinary bladder presenting as a mandibular gingival swelling. AB - Oral cavity metastases mostly originate from the breasts, lungs, or kidneys. Transitional cell carcinoma (TCC), the most frequent malignant tumor of the urinary bladder, rarely metastasizes to the jaws. To the best of our knowledge, only 8 cases of bladder carcinoma have been reported in the English literature to metastasize to the jawbones. A new case of mandibular metastasis of urinary bladder TCC with extension to the gingiva is presented in a 64-year-old white man. The patient was referred for a periodontal infection of the upper right first molar. The clinical examination also showed a gingival swelling located in the lower left premolar region with a hypoasthesia of the left side of the lower lip. The gingival mass was biopsied, and the microscopy showed a mandibular metastatic TCC of the urinary bladder extending to the gingiva. Periodontists should be aware that, although gingival metastases are rare, when they occur they may mimic other local benign pathological conditions. PMID- 11394407 TI - Sucralfate: a help during oral management in patients with epidermolysis bullosa. AB - BACKGROUND: Epidermolysis bullosa (EB) is a group of genetic disorders that lead to blister formation at variable depths in skin and mucosa. Vesicles may arise spontaneously or be caused by friction or trauma. Oral tissue fragility and blistering is common in all EB types. The majority of patients with mild forms of EB are able to receive dental treatment. The prevention of dental caries is most challenging in subjects with severe mucosal involvement, as they are least able to routinely undergo correct preventive procedures. The aim of this study was to evaluate the effectiveness of a sucralfate suspension in reducing both pain and the number of blisters in patients with EB, and to obtain improved oral hygiene and a lower incidence of caries. METHODS: Five patients with dystrophic EB were treated with sucralfate suspension for the prevention and management of oral blisters. Oral blisters were assessed using a quantitative scale, while pain was assessed using visual analogue scale (VAS), and hygiene was evaluated through plaque and gingival indexes. RESULTS: The number of blisters, oral pain, and plaque decreased in all cases. CONCLUSIONS: Oral prophylaxis with sucralfate prevented oral blisters and oral discomfort. The procedure proved to be cost effective and easy to administer. It did not show significant side effects and may be used routinely in patients with EB. PMID- 11394408 TI - Clinical application of a new compact computed tomography system for evaluating the outcome of regenerative therapy: a case report. AB - BACKGROUND: The limitations of conventional 2-dimensional (2-D) radiographs have been emphasized in many studies. Because the image is a 2-D map of the 3 dimensional (3-D) tooth and periodontal tissue, these structures may be superimposed on each other. Recently, 3-D image analysis by computed tomography (CT) was introduced to dentistry, but it has been used only rarely in the periodontal field. METHODS: The purpose of this case report was to introduce the clinical application of a newly developed, compact computed tomography system for evaluating the outcome of regenerative therapy in an adult patient with periodontitis (male, aged 55 years). To evaluate the bone defects including furcation involvement, we took an intraoral radiograph and performed CT scanning on tooth #19. We recorded a number of clinical parameters at the time of surgery. We compared the images of the bone defect obtained by CT images and the conventional radiograph with the true bone defect morphology at the surgical site. RESULTS: One year after the periodontal surgery, we again took CT scans and a conventional radiograph, and the new images of the bone defect were compared and evaluated. The new CT images were reconstructed for 3-D evaluation and compared with those taken before surgery. The CT provides 3-D images of excellent quality for evaluating the morphology of the periodontal bone defect and furcation involvement. CONCLUSIONS: We consider that CT is a useful aid in periodontal examination and diagnosis, and in the prediction and evaluation of periodontal treatment outcomes. PMID- 11394409 TI - [The 71st annual meeting of the Japanese Society for Hygiene. Fukushima, Japan. April 27-30, 2001. Abstracts]. PMID- 11394410 TI - Proceedings of the 10th Hamburg Symposium on Tumor Markers. 5-7 December 1999, Hamburg, Germany. PMID- 11394411 TI - Resolution of four pectate lyase structural genes of Erwinia chrysanthemi (EC16) and characterization of the enzymes produced in Escherichia coli. PMID- 11394412 TI - Strange murder of William of Norwich, 1144. PMID- 11394413 TI - Roles of line stimulation-induced virtual electrodes and action potential prolongation in arrhythmic propagation. PMID- 11394414 TI - Designing a questionnaire. Send a personal covering letter. PMID- 11394415 TI - Vesicovaginal fistula curettage. PMID- 11394416 TI - Influence of hormones on urinary tract infections. PMID- 11394417 TI - Pelviureteric varices. PMID- 11394418 TI - Cervical lymphadenopathy and urinary tract infection. PMID- 11394419 TI - Blind-ending bifid ureter. PMID- 11394420 TI - Hand-assisted laparoscopic surgery--HALS. PMID- 11394421 TI - Current concepts of pelvic congestion and chronic pelvic pain. AB - Chronic pelvic pain in women is a common and disabling illness caused by numerous organic pathologies usually accompanied by varying psychological dysfunctions. Many patients may receive misdiagnosis, misdirected therapies, or do not seek help at all. Pelvic congestion may be responsible for pain in patients without more common diseases, such as endometriosis and pelvic adhesions, among others. Our view of this condition is evolving. In the United States, this medical condition remains controversial. More recent research from the United Kingdom has caused a fresh look at the diagnosis and treatment of chronic pelvic pain produced by pelvic congestion. Potentially, many patients may benefit from a reconsideration of this approach. PMID- 11394422 TI - Long-term follow-up of a controlled trial of laser laparoscopy for pelvic pain. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to assess the long-term efficacy of laparoscopic laser surgery in the treatment of painful pelvic endometriosis. METHODS: We conducted a long-term follow-up of 56 patients who had participated in a randomized, double-blind controlled study at a tertiary referral center for the laparoscopic treatment of endometriosis. The patients had pelvic pain, minimal-to-moderate endometriosis, and underwent laser laparoscopy. We asked patients whether they had now achieved satisfactory symptom relief or whether they had received any further medical intervention for their endometriosis. The main outcome measure was continued symptom relief after treatment and subsequent medical history. RESULTS: Of the original 56 patients, we were able to contact 38 (67.9%). The mean (range) time since operation was 73 months. Painful symptoms had recurred in 28/38 (73.7%) patients at some point since their operation. The median (range) time for recurrence was 19.7 (5-60) months. At the time of follow-up, satisfactory symptom relief was reported in 21/38 (55.3%) patients. The remaining 17/38 (44.7%) patients continued to experience painful symptoms, and eight eventually had a hysterectomy. CONCLUSIONS: This study suggests that operative laparoscopy can have long-term benefits for the majority of women with pelvic pain due to endometriosis, but because of the small numbers, this study lacks the power to demonstrate this conclusively. PMID- 11394423 TI - Hospital resources used for ectopic pregnancy treatment by laparoscopy and methotrexate. AB - OBJECTIVE: To compare resources used in the medical and laparoscopic treatment of unruptured ectopic pregnancy. METHODS: We prospectively recorded all the medical resources required in the treatment of unruptured ectopic pregnancy. The study period ranged from January 1, 1995 to June 30, 1998. Single-dose intramuscular methotrexate injections were administered in 55 women (group I). This therapeutic option was provided on an outpatient basis in small EP (beta-HCG level < 5000 IU/L and hematosalpinx diameter < 3 cm). Serial clinical controls and biologic tests were performed until bHCG became negative. Forty women underwent a laparoscopic salpingostomy because they refused the methotrexate regimen or had "social" contra-indications (ie, predictable difficulties in the follow-up) (group II). Twenty-one patients underwent conservative laparoscopic treatment because of "medical" contraindications to methotrexate (group III). We recorded the resources used with the outpatient and inpatient treatment in each group (methotrexate consumption, operating room acts, length of hospital stay, clinical examinations, biological tests, and sonograms during the follow-up). RESULTS: We observed similar cure rates in each group, but it took significantly longer for beta-HCG to become negative in group 1. However, hospitalization was significantly less often required, and the length of hospital stay was shorter in this group. But length of follow-up, number of office visits, biological tests, sonograms, and subsequent readmissions were significantly more frequent after methotrexate. Despite more severe clinical presentations for patients in group III, we didn't find any significant differences in the hospital resources used in this group in comparison with those used in group II. CONCLUSIONS: The outpatient methotrexate option may result in low consumption of resources for a hospital because most of the follow-up can be performed by city practitioners and laboratories. For the laparoscopic option, efforts should be made to reduce the postoperative hospital stay. PMID- 11394424 TI - Alternative operative techniques in laparoscopic Roux-en-Y gastric bypass for morbid obesity. AB - The only effective treatment for patients with morbid obesity is surgery. Laparoscopic bariatric surgery has become quite popular in attempts to decrease the morbidity associated with laparotomy. In this article, we describe the technical details of laparoscopic Roux-en-Y gastric bypass with three different techniques for creating the 15-cc gastric pouch. These techniques avoid upper endoscopy for the transoral introduction of the 21-mm circular stapler anvil down to the gastric pouch. PMID- 11394425 TI - The role of diagnostic tests in therapeutic choices for gastroesophageal reflux disease. AB - Gastroesophageal reflux disease has different clinical presentations that require different diagnostic and therapeutic approaches. This paper describes the appropriate use of diagnostic tests before and after treatment. Each diagnostic tool is examined from a practical point of view to determine the information it can provide and its possible pitfalls, and to comment on how it can influence therapeutic choices. Performing a preoperative diagnostic evaluation is especially stressed, so as not to select the wrong patient or the wrong procedure. Finally, failures of surgery are examined to understand their causes and to prevent them. The value of the most relevant examinations for diagnosing the causes of failures and choosing the appropriate solution are discussed. PMID- 11394426 TI - Ondansetron versus dehydrobenzoperidol and metoclopramide for management of postoperative nausea in laparoscopic surgery patients. AB - BACKGROUND: In this prospective, randomized, double-blind study, we compared the efficacy of ondansetron versus dehydrobenzoperidol (droperidol) or metoclopramide in the treatment of established postoperative nausea and vomiting in 200 adult patients undergoing laparoscopic surgery under general anesthesia. METHODS: One hundred seventy-three American Society of Anesthesiologists (ASA) I and II patients satisfied inclusion criteria. Fifty-seven patients received ondansetron 4 mg (group O), 57 patients were given droperidol 1.25 mg (group D), and 59 patients received metoclopramide 10 mg (group M). Antiemetic efficacy was compared at 10 minutes and 30 minutes after the administration of the study drug. RESULTS: At 10 minutes, nausea scores in group O dropped from 8.3 to 3.7, in group D from 8.5 to 5, and in group M from 8.4 to 6.7; (P < 0.05 between the three groups). At 30 minutes, nausea scores were 1.3 in group O, 1.7 in group D, and 5 in group M; (P < 0.05 between group M and the other two groups). In the droperidol group, 25% of patients developed sedation. Patient satisfaction was best with ondansetron. CONCLUSIONS: Both ondansetron and droperidol were more effective in the treatment of established postoperative nausea and vomiting than was metoclopramide. However, patients were satisfied best with ondansetron, which acts faster and causes less sedation than droperidol. PMID- 11394427 TI - Laparoscopic management of adnexal masses. AB - BACKGROUND AND OBJECTIVE: Although laparoscopic surgery for removal of adnexal masses is common, controversy exists about the safety and efficacy of this procedure for patients with malignancies. The aim of this study was to evaluate the effectiveness and safety of laparoscopic surgical treatment for patients with adnexal masses. METHODS: This was a retrospective chart review of one surgeon's experience in managing patients diagnosed with adnexal masses at 2 urban referral teaching hospitals in New York City. We reviewed the charts for 100 consecutive patients who underwent operative laparoscopy for management of adnexal masses between March 4, 1996 and November 9, 1998. Conversion to laparotomy, malignancy rate, complications, length of stay, and blood loss were recorded for each patient. RESULTS: Laparoscopic management was successfully completed for 81 of the 100 patients in this study; however, 19 required conversion to laparotomy. All 81 patients managed laparoscopically had a benign diagnosis, whereas 7 of the 19 patients who underwent laparotomy were diagnosed with malignancy. The median length of stay, estimated blood loss, and operating room time were significantly lower for those treated by laparoscopy alone compared with those converted to laparotomy (2 vs. 7 days; 100 vs. 500 ccs; 130 vs. 235 minutes, respectively; P < 0.05). Though few patients were in the laparotomy group, that data are presented for completeness. A total of 10 complications occurred, 4 in the group of patients managed laparoscopically (2 enterotomies, 1 pneumothorax, and 1 vaginal cuff cellulitis). Six complications occurred in those managed with laparotomy (2 enterotomies, 2 wound infections, 1 pneumonia, and 1 postoperative fever). The indications for conversion to laparotomy were: 7 malignancies (5 ovarian cancers and 2 uterine cancers), 7 dense adhesions, 2 small bowel enterotomies, 1 intraoperative bleeding, 1 secondary to a large uterus (880 grams), and 1 secondary to a large myoma (13 cm x 14.5 cm x 6 cm). CONCLUSIONS: The laparoscopic approach is effective and safe for managing patients with adnexal masses of unknown pathology. Malignancies can be diagnosed accurately, converted to laparotomy, and staged appropriately. Adequate surgical skills along with timely use of frozen sections are required for successful operative management. PMID- 11394428 TI - Formidable challenges to teaching advanced laparoscopic skills. AB - Despite the acceptance of laparoscopy for performing routine operations, a need still exists for experienced surgeons and surgical residents to maintain and refine essential surgical skills. Unless used on a frequent basis, laparoscopic skills are not easily maintained. In addition, when new laparoscopic instruments are introduced, surgeons need a way to practice using them that does not involve immediate patient contact. Novice surgeons need the most training of all and ideally would be best served using a standardized teaching curriculum that would cover as many of the basic laparoscopic parameters as possible. This article discusses how best to set up a laparoscopic simulation training program that covers as much ground as necessary, while respecting the restraints of time limitations and monetary concerns. PMID- 11394430 TI - Hand-assisted laparoscopy for the removal of an esophageal leiomyoma. AB - The patient is a 39-year-old male who presented with noncardiac chest pain. His evaluation disclosed an esophageal leiomyoma. In this paper we will demonstrate the pre-operative findings and technique for removal of a benign esophageal tumor using hand-assisted laparoscopy. The patient was discharged home 2 days postoperative and returned to work 2 weeks postoperative with complete resolution of his symptoms. Hand-assisted laparoscopy provides a postoperative course that parallels the recovery from conventional laparoscopy. Additionally, the tactile sense that a surgeon looses from conventional laparoscopy is regained by this technology. PMID- 11394429 TI - Acute cholecystitis: video-laparoscopic versus traditional treatment. AB - It has been shown that a video-laparoscopic approach is the preferred method for treatment of cholecystitis. However, when we consider acute cholecystitis, many questions must be answered. The aim of this study is to compare video laparoscopic and conventional surgery in the management of acute cholecystitis. PMID- 11394431 TI - Rare complications of endoscopic retrograde cholangiopancreatography: two case reports. AB - BACKGROUND: Endoscopic retrograde cholangiopancreatography (ERCP) is a diagnostic procedure with several known risks. We present two rarely reported complications of ERCP and sphincterotomy: transverse mesocolon disruption with ischemic colitis and splenic rupture. RESULTS: The first patient, a 54-year-old female, presented one day following ERCP and stent revision for pancreas divisum. She presented with hypotension and abdominal distention. An abdominal computed tomography (CT) showed a ruptured spleen, which was confirmed on laparotomy. She had a complicated postoperative course and died of multiple organ failure. The second patient is a 56-year-old female who presented five days after ERCP and sphincterotomy with abdominal pain, abdominal wall ecchymosis, and decreasing hematocrit. Her evaluation included hospital admission and abdominal CT scan, which showed free fluid and a large hematoma in the transverse mesocolon. These findings were confirmed on laparotomy and a devascularized segment of bowel was resected. CONCLUSION: Only 6 cases of ERCP-related splenic injury have been reported in the literature. One additional report is available of a fatal splenic artery injury. No previous reports exist of a mesenteric hematoma resulting in bowel devascularization. Prompt evaluation and awareness of potential complications should help capture potentially life-threatening sequelae of ERCP. PMID- 11394432 TI - Laparoscopic hernia repair and bladder injury. AB - BACKGROUND: Bladder injury is a complication of laparoscopic surgery with a reported incidence in the general surgery literature of 0.5% and in the gynecology literature of 2%. We describe how to recognize and treat the injury and how to avoid the problem. CASE REPORTS: We report two cases of bladder injury repaired with a General Surgical Interventions (GSI) trocar and a balloon device used for laparoscopic extraperitoneal inguinal hernia repair. One patient had a prior appendectomy; the other had a prior midline incision from a suprapubic prostatectomy. We repaired the bladder injury, and the patients made a good recovery. CONCLUSION: When using the obturator and balloon device, it is important to stay anterior to the preperitoneal space and bladder. Prior lower abdominal surgery can be considered a relative contraindication to extraperitoneal laparoscopic hernia repair. Signs of gas in the Foley bag or hematuria should alert the surgeon to a bladder injury. A one- or two-layer repair of the bladder injury can be performed either laparoscopically or openly and is recommended for a visible injury. Mesh repair of the hernia can be completed provided no evidence exists of urinary tract infection. A Foley catheter is placed until healing occurs. PMID- 11394433 TI - The Filshie clip for laparoscopic adnexal surgery. AB - BACKGROUND: Gynecologic endoscopic procedures are increasingly common and require the ability to control large vascular structures. METHOD: The Filshie clip is a silicone-lined, titanium occlusive device, originally designed and Food and Drug Administration (FDA) approved for surgical contraception. This device also has the potential for occluding vascular structures during laparoscopic surgery. EXPERIENCE AND RESULTS: We describe a salpingectomy, an excision of bilateral hydrosalpinges, and a salpingo-oopherectomy. We performed all procedures laparoscopically using this device as the primary modality for assuring hemostasis. CONCLUSION: The Filshie clip is a useful and economical device for assuring hemostasis during gynecologic endoscopic surgery. PMID- 11394434 TI - Acalculous cholecystitis in a two year old. AB - The patient is a 2-year-old Caucasian boy with acute acalculous cholecystitis (AC) but none of the predisposing factors that are typically found in patients with this disease. The presentation and clinical course of the disease was typical of AC. Nonsurgical intervention resulted in resolution of the child's initial symptoms. After recurrent bouts of biliary colic over the ensuing ten weeks, further evaluations were completed. Persistent inflammation of the gallbladder was seen on computerized tomographic scans and a nonfunction of the gallbladder was demonstrated through radio-nucleotide scanning. After discussing the findings with the parents, we performed a routine laparoscopic cholecystectomy on the child. The typical presentation, diagnosis, and pathogenesis of AC are discussed. PMID- 11394435 TI - Extrapleural thoracoscopic anterior spinal fusion: a modified video-assisted thoracoscopic surgery approach to the pediatric spine. AB - Video assisted thoracoscopic surgery (VATS) has recently been developed as an alternative to thoracotomy for anterior spinal surgery. We report a case in which an extrapleural dissection was combined with VATS to further improve this approach. PMID- 11394436 TI - The laparoscopic retrieval of an orthopedic fixation pin from the liver with repair of an associated diaphragmatic laceration. AB - We report the successful removal of a shoulder fixation pin from the right lobe of the liver with intracorporeal repair of a diaphragmatic laceration. An expeditious workup and urgent operative intervention were required. We adhered to the principles of room setup, optical correctness, establishment of the triangle of success, appropriate instrument entry and convergence angles, two-handed surgical skills, and competence in intracorporeal suturing techniques that were all required for successful completion of the case. We also present a review of the significant literature. PMID- 11394438 TI - Transcriptional regulation of c-fms gene expression. PMID- 11394437 TI - Video-assisted parathyroidectomy by a skin-lifting method for primary hyperparathyroidism. AB - OBJECTIVE: The use of endoscopic surgical procedures has rapidly spread to abdominal and thoracic surgeries and subsequently to surgeries of the neck region. Several surgeons initiated endoscopic parathyroidectomy using CO2 insufflation to create the working space; however, they reported various complications. We describe here a skin-lifting method that may have few complications. METHODS: A 65-year-old man was diagnosed with primary hyperparathyroidism due to a solitary adenoma of the left inferior parathyroid gland. A 3-cm oblique incision was made below the left clavicle, and a 5-mm incision was made on the lateral neck. After the skin was lifted up, we performed video-assisted parathyroidectomy. RESULTS: Parathyroid extirpation took 2 hours and blood loss was minimal. The patient had minimal pain and no complications postoperatively. Serum concentrations of calcium and intact parathyroid hormone were normalized on the next day. CONCLUSION: Using the skin-lifting method, we obtained a sufficient operative view and encountered no complications. This procedure is cosmetically desirable, and we consider it a feasible alternative for the treatment of parathyroid adenoma. PMID- 11394439 TI - The quartz crystal microbalance as a novel means to study cell-substrate interactions in situ. AB - The quartz crystal microbalance (QCM) was first introduced as a mass sensor in gas phase and in vacuum. Since oscillator circuits capable of exciting shear vibrations of quartz resonators under liquid loading have been developed, the QCM became accepted as a new, powerful technique to follow adsorption processes at solid-liquid interfaces in chemical and biological research. Lately, the QCM technique has attracted considerable interest as a novel means to monitor cell substrate interactions of mammalian cells in vitro. Because the establishment and modulation of cell-substrate contacts is important for many physiological processes, and potent techniques to measure these interactions noninvasively are rare, the present review highlights applications of the QCM technique in this field. The suitability of the QCM device to monitor attachment and spreading of mammalian cells in real time has been well established. The QCM response is dependent on the individual cell type that is examined. In order to identify the sources for these cell-type-specific results of QCM readings, and to understand the information content of the signal, attempts have been made to decompose the overall QCM response into subcellular contributions. The aforementioned subjects, together with a condensed introduction into the QCM technology, are included in this article. PMID- 11394440 TI - Negative regulation of Janus kinases. AB - The precise regulation of both the magnitude and the duration of Janus kinase (JAK) catalytic activity is essential for the cytokine orchestration of many biological processes, and the dysregulation of JAK activity has pathological implications. Immunosuppressive disease states, such as X-linked severe combined immunodeficiency, arise from inappropriate JAK inhibition. In contrast, a limited number of cancers, primarily leukemias, result from constitutive or enhanced activation of JAK activity. JAKs are no longer implicated only in classic cytokine receptor-mediated signaling pathways, but are now also known to integrate indirectly into other receptor-mediated signal transduction processes. Therefore, an increasing number of therapeutic applications exist for biological response modifiers that can restore aberrant JAK activity to normal levels. Exciting breakthroughs in both physiological and pharmacological methods of selective inhibition of cytokine-JAK-signal transducers and activators of transcription pathways have recently emerged in the form of suppressors of cytokine signaling (also known as cytokine-inducible SH2 protein, JAK-binding protein, or STAT-induced STAT inhibitor) proteins and novel dimethoxyquinazoline derivatives, respectively. The basis of these and other mechanisms of negative regulation of JAK activity, including the suppression of jak expression levels caused by tumor- or pathogen-derived agents, the complex interactions of JAKs with phosphatases, and the redox regulation of JAK catalytic activity, is the focus of this review. PMID- 11394441 TI - Tumor suppressor gene regulation of cell growth: recent insights into neurofibromatosis 1 and 2 gene function. AB - The development of cancer involves a myriad of genetic changes that impact on multiple processes important for the orderly regulation of cell growth and differentiation. Genes whose protein products are disrupted during neoplastic transformation are termed "tumor suppressor genes" (TSGs). Many of these TSGs are associated with familial cancer predisposition syndromes, in which affected individuals have an increased risk of certain malignancies. Studies on the mechanism of action for known TSGs have revealed three intracellular loci of critical importance: environmental sensing and signal initiation, signal propagation and transduction, and cell cycle control. The neurofibromatosis 1 and neurofibromatosis 2 genes are discussed as illustrative examples of tumor suppressors that function at the levels of signal transduction and environmental sensing, respectively. PMID- 11394442 TI - Determinants of G protein inhibition of presynaptic calcium channels. AB - The modulation of presynaptic calcium (Ca) channels by heterotrimeric G proteins is a key factor for the regulation of neurotransmission. Over the past 20 yr, a significant understanding of the molecular events underlying this regulation has been acquired. It is now widely accepted that binding of G protein betagamma dimers directly to the cytoplasmic region linking domains I and II of the Ca channel alpha1 subunit results in a stabilization of the closed conformation of the channel, thereby inhibiting current activity. The extent of the inhibition is dependent on the Gbeta subunit isoform, and is antagonized by both strong membrane depolarizations and protein kinase C-dependent phosphorylation of the channel. Finally, the inhibition is critically modulated by regulator of G protein signaling proteins, and by proteins forming the presynaptic vesicle release complex. Thus, the regulation of the activities of presynaptic Ca channels is becoming increasingly complex, a feature that may contribute to the overall fine-tuning of Ca entry into presynaptic nerve termini, and thus, neurotransmission. PMID- 11394443 TI - The effect of secondary structure on cleavage of the phosphodiester bonds of RNA. AB - This review discusses the effects the secondary structure of an RNA molecule has on the inherent reactivity of its phosphodiester bonds, and on the catalytic activity of metal ion-based cleaving agents. The basic principles of the intramolecular transesterification of RNA phosphodiester bonds, particularly cleavage, are first briefly described. Studies of the structural effects on the cleavage, in the absence and in the presence of metal ion catalysts, are then reviewed, and the sources of the reactivity differences observed in different structures are discussed. PMID- 11394444 TI - Laparoscopic approach to adrenal carcinoma. AB - There is general agreement on the suitability of the laparoscopic approach for benign adrenal lesions but controversy about using laparoscopy for suspected adrenal malignancy. This article reviews the literature on laparoscopic adrenalectomy for cancer: the operative techniques and indications and contraindications. PMID- 11394445 TI - Laparoscopic radical/total nephrectomy: a decade of progress. AB - The first laparoscopic radical/total nephrectomy for a renal tumor was performed in June 1990. Since that time, the procedure has evolved as numerous surgeons have contributed novel strategies and technical advances. The state of the art is reviewed, including transperitoneal laparoscopic and hand-assisted techniques, as well as the retroperitoneal approach. Operative and postoperative data are reviewed with the goal of determining four factors: the efficacy, efficiency, morbidity, and cost of the procedure. Within the limits of available follow-up for this novel procedure, it appears to be as effective as open surgery in rendering the patient tumor free. Although it clearly is a less painful and less disabling procedure than open surgery, our understanding of the efficiency of the laparoscopic procedure remains in flux. The operative times for laparoscopic radical/total nephrectomy are approaching those of traditional open radical nephrectomy, although intraoperative costs remain higher and thus must be balanced against decreased hospitalization and convalescence. PMID- 11394446 TI - Results of retroperitoneal laparoscopic radical nephrectomy. AB - PURPOSE: To analyze the retroperitoneal approach to laparoscopic radical nephrectomy in terms of feasibility, safety, morbidity, and cancer control. PATIENTS AND METHODS: We reviewed the records of 50 consecutive patients with renal cancer underwent radical nephrectomy via the retroperitoneal laparoscopic approach from 1995 through 1999. RESULTS: The mean operative time was 139 minutes (range 60-330 minutes) with a mean of 149.78-mL operative blood loss (0-1500 mL). The mean renal size was 100 mm (70-150 mm) with a mean tumor size of 38.6 mm (20 90 mm). The postoperative hospital was 6 days (2-13 days). Three open conversions were necessary: one for laparoscopically uncontrolled bleeding and two because obesity interfered with surgery. We noted two major complication and two minor complications. Two disease progression have been noted to date. One patient with a pT3 grade 2 renal-cell carcinoma had a local recurrence with liver metastasis 9 months after the procedure and died 19.7 months after radical nephrectomy. Another patient with a pT3aN+M+ cancer died 23.1 months after the procedure. CONCLUSION: Retroperitoneal laparoscopic nephrectomy for kidney cancer requires further assessment. It seems to have several advantages over open radical nephrectomy and to be effective and safe for small (<50-mm) renal tumors. PMID- 11394447 TI - Cryosurgery and needle ablation of renal lesions. AB - Laparoscopic renal cryoablation is a minimally invasive alternative for treating renal tumors utilizing narrow probes cooled with a compressed gas such as argon or carbon dioxide. At this time, cryotherapy has shown the most promise as an alternative to partial nephrectomy as a nephron-sparing treatment for renal tumors. Radiofrequency ablation employs needle electrodes placed percutaneously directly into renal lesions to deliver energy, creating high temperatures leading to cell death. High-intensity focused ultrasound is a noninvasive technique in which focused ultrasound energy is applied to cause cell death within the focal zone. Microwave thermotherapy uses small applicators to deliver microwave energy to tissues, resulting in the generation of heat. Although RF, HIFU, and microwave thermotherapy show promise as energy sources for tumor ablation, they are in the early stages of development. Little is known about their acute and chronic histologic effects and long-term efficacy as a treatment for malignant disease. Further work is needed to develop cryosurgery and needle ablation in order to delineate what role these techniques will ultimately play in the management of RCC. PMID- 11394448 TI - Laparoscopic partial nephrectomy and wedge resection for the treatment of renal malignancy. AB - The widespread use of abdominal ultrasonography, CT, and MRI has led to an increase in the number of incidentally detected renal masses, some of which are malignant. Numerous studies suggest that partial nephrectomy or wedge resection of these lesions yield cure rates similar to those obtained with radical surgery. Laparoscopic nephron-sparing surgery is one of the more challenging minimally invasive surgical techniques, and its use is largely restricted to specialized medical centers. The techniques and available results are described. PMID- 11394449 TI - Transitional-cell carcinoma of the renal pelvis: ureteroscopic and percutaneous approach. AB - There are a variety of publications advocating the ureteroscopic or the percutaneous approach for the treatment of transitional cell carcinoma of the renal pelvis. The diagnostic tool of choice for the upper urinary tract and collecting system is the flexible ureteroscope. One of the major concerns about ureteroscopic management of renal disease initially was the lack of flexibility of the instruments and therefore the inability to deal with demanding sites. The advent of new ureteroscopic techniques, as well as the continuous evolution of the technology, have paved the way for safe and effective access to the upper urinary tract. In the hands of an experienced urologist, such procedures can provide reliable treatment options for small upper urinary tract lesions. Coupling minimal morbidity with ever-improving optics and flexibility, the ureteroscope of today leaves no area of the urinary tract unseen. In patients with bulky tumors or in whom easy access and resection is not possible ureteroscopically, the percutaneous approach to the renal pelvis, although more invasive, provides a better working environment. Clearly, the most difficult aspect of ureteroscopic access to the lower pole is not just visibility but the loss of deflection caused by passage of various instruments through the working channel. Direct access via percutaneous approach with a large resectoscope avoids these problems. PMID- 11394450 TI - Retroperitoneal laparoscopic nephroureterectomy and management options for the distal ureter. AB - Radical nephroureterectomy, including en bloc excision of the ureter with a bladder cuff, is the standard treatment for upper tract transitional-cell carcinoma (TCC). This procedure requires either a flank and lower abdominal incision or an extended flank incision. Laparoscopic surgery for TCC has been used at several medical centers; the most challenging and controversial aspect is the oncologically correct management of the distal ureter. We believe that the Cleveland Clinic technique of securing the distal bladder cuff intravesically while simultaneously occluding the distal ureter prevents tumor spillage and allows accurate and complete resection of the targeted ureter in a manner mirroring the open procedure. PMID- 11394451 TI - Hand-assisted laparoscopic nephroureterectomy versus open nephroureterectomy for the treatment of transitional-cell carcinoma of the upper urinary tract. AB - BACKGROUND AND PURPOSE: For patients with upper tract transitional-cell carcinoma (TCC), nephroureterectomy with removal of a bladder cuff is the standard of care. Historically, it has been performed using two incisions or one large incision extending from the lateral flank to the symphysis pubis. We describe an alternative using endoscopic management of the bladder cuff combined with hand assisted laparoscopic (HAL) nephroureterectomy. We compared our results using these minimally invasive advances with those of a contemporary open nephroureterectomy series. PATIENTS AND METHODS: Between May 1998 and June 1999, we performed 11 HAL nephroureterectomies with endoscopic management of the bladder cuff for the treatment of upper tract TCC. The results were compared with those in a contemporary series of 11 patients undergoing the traditional open operation at our institution. The patient age, male:female ratio, and ASA classification were similar in the two groups. Intraoperative measures considered were operative time, estimated blood loss, need for transfusion, complications, specimen weight and volume, pathologic stage and grade of the tumor, and the status of the surgical margins. Postoperative endpoints were time to sustained fluid intake; epidural, parenteral, and oral narcotic requirements; length of stay; and complications. Follow-up, specifically disease recurrence and overall survival, was recorded. RESULTS: The mean operative time was 291 minutes for HAL v 232 minutes for the open operation (P = NS). The average blood loss was 144 v 311 mL (P = 0.04), the mean specimen weight 368 v 392 g (P = NS), and the mean specimen volume was 630 v 693 cc (P = NS). No patient in the HAL group had a positive surgical margin, but one patient in the open surgery group did. The time to sustained fluid intake postoperatively averaged 1.4 v 2.3 days for the HAL and open groups, respectively (P = NS). The epidural narcotic requirement was 0 v 2.7 days (P < 0.001), the mean parenteral narcotic requirement was 45 v 44 mg of morphine sulfate equivalent (P = NS), and the oral narcotic requirement was 5.8 v 16 tablets (P < 0.04). The average length of stay was 4.6 days for the HAL group v 6.1 days for the open group (P = 0.04). In both groups, 7 of the 11 patients (63%) were without evidence of disease with a mean follow-up of 13 (HAL) and 17 (open) months. CONCLUSIONS: Hand-assisted laparoscopic nephroureterectomy with endoscopic management of the bladder cuff is an efficacious alternative to open surgery. The operative time, specimen weight and size, and risk of recurrence for the two procedures are similar. However, convalescence, as measured by pain medication requirements and length of stay, is significantly better with laparoscopy. Longer follow-up with larger numbers of patients is in progress. PMID- 11394452 TI - Ureteroscopic surgery for upper tract transitional-cell carcinoma: complications and management. AB - In some patients with small low-grade transitional-cell carcinoma of the ureter, the lesion can be treated ureteroscopically with the laser or electrocoagulation. Patients must be compliant with lifelong frequent surveillance. Surgical complications such as ureteral perforation and stricture are uncommon. The authors provide advice on technique and on preventing and managing complications. PMID- 11394453 TI - When is partial ureterectomy acceptable for transitional-cell carcinoma of the ureter? AB - Because the disease is so often multifocal and has a high likelihood of recurrence, transitional-cell carcinoma of the ureter traditionally has been treated by nephroureterectomy. As reviewed in this article, partial ureterectomy may be a feasible option for some patients, particularly those with a solitary kidney, bilateral tumors, or renal insufficiency. This conservative procedure also may be appropriate for patients with grade 1 and possibly grade 2 tumors in stage Ta/T1. PMID- 11394454 TI - Videoendoscopically assisted transvaginal radical cystectomy. AB - We describe our experience of anterior pelvic exenteration for bladder cancer in women using a combined transvaginal and laparoscopic approach. The feasibility of videoendoscopically assisted transvaginal cystectomy was demonstrated several years ago, but the indications have been greatly reduced by the spreading use of orthotopic neobladders in women. Moreover, nulliparous patients or patients with vaginal atresia are not suitable for this technique, even if the specimen can also be retrieved through the minilaparotomy used for performing urinary diversion. In the last 3 years, we have performed only four additional cases. Nevetheless, the results are satisfactory. We did not have any major intraoperative complication. Patients were discharged after 7 to 11 days (average 8.6 days) without any postoperative complications. The minimum survival was 13 months, and four patients are still alive. The operation itself is not easy and therefore can be offered only by centers where videoendosurgery has already entered common clinical practice. PMID- 11394455 TI - Laser treatment for invasive and noninvasive carcinoma of the bladder. AB - Lasers are widely used in urologic surgery and have many well-developed applications. The use of lasers for the treatment of bladder cancer has been proven to be safe and minimally invasive. The neodymium:YAG laser is the most widely used instrument, although the holmium:YAG laser has also gained recent popularity. Noninvasive, small lesions are especially amenable to management with laser energy and have been treated with success rates that are at least as good as those of standard electrocautery resection. Complication rates are low. Laser therapy of invasive bladder cancer has significant limitations and should probably be restricted to stage T2 lesions. PMID- 11394456 TI - Radical transurethral resection in the management of muscle-invasive bladder cancer. AB - The morbidity of radical cystectomy and early reports of good results have stimulated interest in radical transurethral resection of bladder tumors (TURBT) for muscle-invasive transitional-cell carcinoma of the bladder. Various investigators have used surgery alone or with adjuvant or neoadjuvant chemotherapy or radiation. Further research is necessary to define the indications, but at present, radical TURBT for muscle-invasive cancer appears to be appropriate for patients too ill to undergo radical cystectomy, those who decline the open operation, and those enrolled in clinical trials of this approach to bladder cancer. PMID- 11394457 TI - Indications, technique, and results of laparoscopic pelvic lymphadenectomy. AB - Despite considerable clinical research, there is still controversy about the optimal management of the pelvic lymph nodes in men with prostate cancer. This article reviews the creation and application of selection criteria for laparoscopic pelvic lymphadenectomy and describes the various techniques. PMID- 11394458 TI - Results and side effects of high-intensity focused ultrasound in localized prostate cancer. AB - At the time of diagnosis, prostate cancer is organ confined in 70% of the cases. A quarter of these patients undergo local therapy (surgery/radiation); 75% risk disease progression by "watchful waiting" or systemic side effects through hormonal ablation. Local high-intensity focused ultrasound (HIFU), as minimal invasive tissue coagulation (85 degrees C), ablates prostatic tissue with high precision. Since April 1996, 184 patients have undergone 232 sessions of transrectal HIFU therapy (average 90 min) under spinal anesthesia at 2.25/3.0 MHz, 50 W, and a penetration depth of 25 mm. The follow-up serum prostate specific antigen (PSA) concentration, sextant biopsies, International Prostate Symptom Score (IPSS), quality of life measures (QoL), and complaint registration provide the foundation for this clinical evaluation. Follow-up sextant biopsies (an average of 1.9) showed 80% of the patients to be cancer free. In men with residual cancer, the tumor mass was reduced more than 90%. The PSA nadir in 97% was <4 ng/mL, including 61% with values <0.5 ng/mL. After primary HIFU, no severe side effects (fistula, second or third grade incontinence, rectal mucosal burns) occurred. All patients had a suprapubic tube (average 29 days), and 33% needed a transurethral debris resection averaging 7 g. They were discharged within 23 hours. According to the short-term follow-up transrectal HIFU enables minimal invasive local prostate tissue ablation with high rates of negative biopsies, low PSA nadir, and low complication rate. PMID- 11394459 TI - Laparoscopic radical prostatectomy: the lessons learned. AB - PURPOSE: To evaluate the operative, oncologic, and functional results of laparoscopic radical prostatectomy based on an initial series of 350 patients. PATIENTS AND METHODS: Between January 1998 and May 2000, 350 consecutive patients underwent laparoscopic radical prostatectomy according to our technique. The study of operative morbidity was based on all intraoperative and postoperative complications. The oncologic assessment was based on clinical, laboratory, and intraoperative and postoperative pathological data. Postoperative functional results were assessed by the ICS-male self-administered questionnaire. RESULTS: No deaths were observed in this series. Conversion was required in seven cases, exclusively among the first 70 patients. The mean operating time was 217 +/- 59 minutes, including the lymphadenectomy phase that was considered necessary in 21.4% of patients, and 195 +/- 56 minutes for the most recent 200 patients. The mean intraoperative blood loss was 354 +/- 250 mL. The overall transfusion rate was 5.7% and 2.8% in the last 250 patients. Intraoperative complications were reported in 14 patients (4%), and the reoperation rate was 3.7%. The mean postoperative bladder catheterization time was 5.8 +/- 3.3 days, and the catheter could be removed before the 5th day in 41% of patients. The mean hospital stay was 6 +/- 3.9 postoperative days (range 2-33 days). By pathologic stage, the positive surgical margin rate was 3.6% for pT2a specimens (3 patients), 14% for pT2b specimens (29 patients), 33% for pT3a specimens (12 patients), and 43.5% for pT3b specimens (10 patients). In the first 75 patients with pT2N0/Nx negative margin specimens and a follow-up of >12 months, the PSA concentrations was <0.2 ng/mL in 92% of patients. The continence rate (no protection necessary either during the day or at night) among the first 133 patients was 85.5% and the postoperative erection rate was 59% among 22 selected consecutive patients. CONCLUSIONS: This study confirms the value, in our experience, of the laparoscopic approach to radical prostatectomy, which allows satisfactory cancer control associated with low perioperative morbidity and encouraging functional results in terms both of continence and erectile function. PMID- 11394460 TI - Laparoscopic retroperitoneal lymph node dissection. AB - PURPOSE: Retroperitoneal lymph node dissection is the most sensitive and specific diagnostic modality for detecting occult lymph node metastases in clinical stage I testicular tumor. In stage II disease, residual tumors after chemotherapy have to be removed surgically. To reduce the morbidity of these procedures we have replaced open surgery by laparoscopy. PATIENTS AND METHODS: Between August 1992 and December 1999 125 patients underwent laparoscopic RPLND (stage I: 76 pts., stage II: 49 pts.) RESULTS: Laparoscopic RPLND could be completed as planned in all but two patients in whom bleeding required conversion to open surgery. Once the learning curve had been overcome, mean operative time decreased significantly from 476 to 219 min for stage I and averaged 226 min in stage IIB disease. Only minor postoperative complications occurred such as asymptomatic lymphoceles (7 pts.) and chylous ascites (6 pts.). Mean post-op hospital stay was 3.3 and 3.5 days, respectively (stages I and II). Mean followup is currently 46 months for stage I and 35 months for stage II tumors. Over this period a single retroperitoneal recurrence was observed (stage I), which, however, was not due to surgical failure, but to false negative histologic findings. All other patients have remained free of relapse. CONCLUSIONS: Laparoscopic RPLND is a demanding procedure with a long and steep learning curve. It has proved feasible also after chemotherapy. The diagnostic accuracy of laparoscopic RPLND was as good as that of the open procedure, while the morbidity is significantly lower. Tumor control was not compromised by the laparoscopic approach. PMID- 11394461 TI - Biomechanical analysis of peak and cumulative spinal loads during simulated patient-handling activities: a substudy of a randomized controlled trial to prevent lift and transfer injury of health care workers. AB - Back injuries are a serious problem for nursing personnel who perform frequent patient-handling activities. Common prevention strategies include body mechanics education, technique training, and ergonomic interventions such as the introduction of assistive equipment. This investigation assessed and compared the effectiveness of two patient-handling approaches to reducing injury risk. One strategy involved using improved patient-handling technique with existing equipment, and the other approach aimed at eliminating manual patient handling through the use of additional mechanical and other assistive equipment. Both intervention arms received training in back care, patient assessment, and use of the equipment available on their particular wards. An analysis of compliance with interventions and the effects of patient-handling methods on both peak and cumulative spinal compression and shear during various tasks was conducted. Results showed greater compliance with interventions that incorporated new assistive patient-handling equipment, as opposed to those consisting of education and technique training alone. In several tasks, subjects who were untrained or non-compliant with interventions experienced significantly higher peak spinal loading. However, patient-handling tasks conducted with the aid of assistive equipment took substantially longer than those performed manually. This, along with variations in techniques, led to increases in cumulative spinal loading with the use of patient-handling equipment on some tasks. Thus, the use of mechanical assistive devices may not always be the best approach to reducing back injuries in all situations. No single intervention can be recommended; instead all patient handling tasks should be examined separately to determine which methods maximize reductions in both peak and cumulative lumbar forces during a manoeuver. PMID- 11394462 TI - Validity of fixed-interval observations for postural assessment in construction work. AB - While observing six simulated construction tasks in the field, trained analysts recorded arm, trunk and leg postures categorically with two fixed-interval observational protocols. Observations were compared to measurements obtained with an electronic postural assessment system coupled with video analysis. The electronic postural assessment system consisted of electronic inclinometers to measure upper arm posture, knee flexion and trunk flexion, coveralls to house the inclinometer wiring, and an eletrogoniometric system to measure trunk lateral bending and twisting. Video analysis included frozen-frame analysis that corresponded to the moment of observation and simulated real-time analysis. Measurements were made on five male participants who each performed three tasks representative of construction laborers' work. Agreement among the observational and reference methods was generally high, although significant differences in measured frequency of exposure existed for knee flexion, trunk lateral bending and trunk twisting. The results suggest that, under appropriate conditions, discrete observations can be used to obtain reasonably accurate estimates of exposure frequency for broad categories of certain body postures. PMID- 11394464 TI - Anthropometry of norwegian light industry and office workers. AB - Several of the Directives and Standards developed by CEN, e.g. those dealing with Machine Safety and Personal Protective Equipment, depend on anthropometric information. For Norway, anthropometric data relevant for these standards have either been lacking or are very old. Two hundred males and 199 female employees of light industry and office work, between 20 and 39 years of age were studied. Body mass plus 29 different body dimensions were measured. Mean age of the total male and female populations were 30.0+/-5.6 and 30.0+/-5.4 years, respectively. Mean stature for males was 1796+/-66 mm, while the corresponding value for females was 1661+/-61 mm. Body weights for the two populations averaged 78.2+/ 11.1 and 63.4+/-9.3 kg, respectively. There were no significant differences in stature between the two age groups, neither in males nor in females. In body mass, males 30-39 years of age had significantly higher body mass than the 20-29 years old, while this was not the case in females. PMID- 11394463 TI - Computer terminal work and the benefit of microbreaks. AB - Microbreaks are scheduled rest breaks taken to prevent the onset or progression of cumulative trauma disorders in the computerized workstation environment. The authors examined the benefit of microbreaks by investigating myoelectric signal (MES) behavior, perceived discomfort, and worker productivity while individuals performed their usual keying work. Participants were randomly assigned to one of three experimental groups. Each participant provided data from working sessions where they took no breaks, and from working sessions where they took breaks according to their group assignment: microbreaks at their own discretion (control), microbreaks at 20 min intervals, and microbreaks at 40 min intervals. Four main muscle areas were studied: the cervical extensors, the lumbar erector spinae, the upper trapezius/supraspinatus, and the wrist and finger extensors. The authors have previously shown that when computer workers remained seated at their workstation, the muscles performing sustained postural contractions displayed a cyclic trend in the mean frequency (MNF) of the MES (McLean et al., J. Electrophysiol. Kinesiol. 10 (1) (2000) 33). The data provided evidence (p < 0.05) that all microbreak protocols were associated with a higher frequency of MNF cycling at the wrist extensors, at the neck when microbreaks were taken by the control and 40 min protocol groups, and at the back when breaks were taken by the 20 and 40 min protocol groups. No significant change in the frequency of MNF cycling was noted at the shoulder. It was determined (p < 0.05) that microbreaks had a positive effect on reducing discomfort in all areas studied during computer terminal work, particularly when breaks were taken at 20 min intervals. Finally, microbreaks showed no evidence of a detrimental effect on worker productivity. The underlying cause of MNF cycling, and its relationship to the development of discomfort or cumulative trauma disorders remains to be determined. PMID- 11394465 TI - The ergonomics of flight management systems: fixing holes in the cockpit certification net. AB - Recent air traffic control regulations mandate the installation of computer-based flight management systems in airliners across Europe. Integrating and certifying add-on cockpit systems is a long and costly process, which in its current form cannot meaningfully address ergonomics aspects. Two levels of problems occur: add on systems carry many "classic" HCI failures, which could easily be addressed with modified certification requirements. Further, adding new technology changes practice, creates new skill and knowledge demands and produces new forms of error, which are more difficult to assess in advance. However, one innovative certification approach for add-on cockpit systems, based on the use of a representative population of user pilots, was found to be promising. This method minimizes the subjective bias of individual pilots in addition to defining pass/fail criteria in an operational environment. PMID- 11394466 TI - Electromyographic and discomfort analysis of confined-space shipyard welding processes. AB - This study examined muscle fatigue and discomfort in a confined-space welding operation at a shipyard. Surface electromyography (SEMG) was recorded from seven upper extremity and torso muscles of welders welding in a mock-up of the work environment. Following spectral transform of the SEMG data the percentage of the total signal power in the 10-30 Hz frequency band was compared over time during welding. For the conventional stick electrode welding process (SMAW) several muscles exhibited an increase in the percent of the total signal power in the low frequency band. Fewer muscles exhibited this fatigue-related spectral density shift with a wire welding process (FCAW) the shipyard has considered adopting. This finding suggests that localized muscle fatigue may be reduced by a change to the wire welding process. Subjectively reported discomfort was generally low for both processes, but confirmed the finding that discomfort in the low back and shoulder regions is experienced in this welding operation. PMID- 11394467 TI - A system for the measurement of grip forces and applied moments during hand tool use. AB - Quantification of the forces applied with or by hand tools can be a difficult but important component of an ergonomic evaluation. This paper describes a device for measuring gripping forces and the moments generated by a hand tool. Laboratory characterization indicated that the device had good linearity (r2 = 0.999) with minimal hysteresis or creep. The working range exceeds 700N for gripping forces, and 28 and 16Nm for the two applied moment axes. The device, configured as a boning knife, was sensitive to differences in grip forces and applied moments in a simulated meat cutting task requiring distinct levels of precision. Significant individual variation in the "efficiency" of grip force was also observed. The system design is flexible, allowing for additional tool configurations. PMID- 11394468 TI - An investigation of how acute muscle pain modulates performance during computer work with digitizer and puck. AB - The purpose was to investigate the influence of muscle pain on work performance during computer work with digitizer and puck. Muscle pain was induced by infusion of hypertonic saline in the trapezius and the extensor carpi ulnaris (ECU) muscles on two separate days. Twelve healthy subjects participated. A computer task was performed in three 6 min sessions: baseline, pain, after pain. The computer task comprised production of drawings at maximal work pace. One drawing was defined as a work cycle. Work cycle time, number of puck button clicks, and screen pixels the cursor had moved per cycle were assessed. Shoulder pain did not influence these variables. Cycle time decreased from 13.8 (SD 2.2) to 13.0 s (SD 1.9) compared to baseline (p < 0.05) during ECU muscle pain. The increased or unchanged performance suggests that acute moderate muscle pain has minor influence on performance during computer work with digitizer and puck. PMID- 11394469 TI - The composition of a graph on the decline of total body strength with age based on pushing, pulling, twisting and gripping force. AB - This study is part of a large project on design-relevant characteristics of ageing users. A total of 750 young and elderly, male and female subjects were tested. All subjects were able to live independently. Differences between the sexes were mapped as well as changes in force with age. The relationships between five different isometric forces (pushing, pulling, twisting and left and right gripping) were investigated. The possibilities of extending information on these forces to other forces and other subject groups were also studied. The results indicate that the different types of force are closely related. A strength score was devised. We conclude that calculation of unmeasured forces by means of a general strength score could be possible. Furthermore, we found that the percentage decrease in strength with age is similar for men and women. PMID- 11394470 TI - A compatible chord code for inputting elements of Chinese characters. AB - A compatible chord code for inputting elements of Chinese characters (ECC) to computer was proposed. It capitalized on the graphic compatibility between ECC and chord combination of keys (CCK) on a single-handed chord keyboard with five keys. Experimental results showed that the proposed compatible chord code was better than a code that randomly mapped ECC onto CCK with respect to learning time and response time. Explicit indication of the graphic compatibility between ECC and CCK did not enhance memorizing the compatible code. PMID- 11394471 TI - Comparison of laboratory studies with predictions of the required sweat rate index (ISO 7933) for climates with moderate to high thermal radiation. AB - For International Standard ISO 7933 (Hot environments-analytical determination and interpretation of thermal stress using calculation of required sweat rate. International Organization for Standardization, Geneva, 1989) it has been questioned whether the heat stress index "required sweat rate" is applicable in environments where mean radiant temperature (t(r)) exceeds dry air temperature (t(a)). Based on a re-analysis of 556 climatic chamber experiments on 16 subjects the observed sweat rates were compared with required sweat rates predicted according to the ISO standard. Under climatic conditions with t(r) > t(a) the predictions overestimate the observed sweat rates on average by 86% in clothed subjects and by 25% in nude subjects. In climates with t(r) = t(a) the predictions highly agreed with the observed sweat rates in nude subjects and were slightly overestimated (13%) in clothed subjects. The misjudgement of the present ISO standard is obviously caused by an inappropriate calculation of radiative heat exchange. Corrections are proposed to improve the validity concerning climates with t(r) > t(a). PMID- 11394472 TI - Prospects of gene therapy for ocular diseases. PMID- 11394473 TI - Is refractive surgery justified? AB - Refractive surgery may be defined as any kind of eye surgery in which attempt is made to produce a level of spectacle error that is accepted to the patient. Cataract surgery with lens implantation is the most often done refractive surgery on the eye. The following kinds of refractive surgery are being practised: With normal crystalline lens, phacic intra-ocular lens, surgery with cornea, using excimer laser, other newer techniques. Radial keratotomy is one procedure of refractive surgery with cornea. In this operation, radial cuts are made on the cornea starting near the edge of the pupil and reaching up to the limbus. Number of cuts vary from 4 to 24 depending upon the seventy of myopia. Its advantage is its cost effectiveness and can be performed in a remote area. There are two ways of excimer laser: photorefractive keratectomy (PRK) and laser assisted in-situ keratomileusis (LASIK). The processes are elaborated in the article. Before any refractive surgery is performed, it is imperative that the whole of the eye receives full and detailed attention. The patient needs to be explained of the procedure. Utmost care should be taken to observe sterilisation of the instruments and the environment. PMID- 11394475 TI - Primary glaucomas: current concepts and management. AB - Glaucoma is a leading cause of blindness. Delay in diagnosis of open angle glaucoma (OAG) results from lack of symptoms. In angle closure glaucoma (ACG) there is often neglect of symptoms. Creating awareness about the disease and screening the high risk groups would reduce the burden of irreversible blindness due to glaucoma. Adequate therapy in the form of drops, lasers and surgery is available. Close interaction with the physician is important as many topical antiglaucoma medications have adverse systemic side-effects and many drugs used for systemic diseases raise the intra-ocular pressure (IOP). All drugs currently available for glaucoma lower IOP but new drugs for neuroprotection may change the future management strategies. While drugs and surgery are the mainstay for OAG, laser iridotomy is the definitive treatment for ACG. PMID- 11394474 TI - Recent developments in vitreoretinal surgery. AB - In the past three decades, a great improvement has occurred in microsurgical techniques used in the management of various eye diseases involving retina and vitreous. Advances in instrumentation has made the surgery easier and refined. The instruments used are narrated widely in this article. Various vitreous substitutes have been developed and are required in vitreoretinal surgery. These are gases, silicone oil, perfluorocarbon liquids and fluorosilicone oil. Anterior segment indications for vitreous surgery are: Vitreous loss during cataract surgery, thick after cataracts that can't be managed with Nd: YAG capsulotomy, vitreocorneal touch, updrawn pupils, incarcerated vitreous in the wound causing cystoid macular oedema, malignant glaucoma, penetrating keratoplasty in aphakic patients, congenital cataracts and filtering procedures in aphakic eyes. In posterior segment indications, vitrectomy is useful in penetrating trauma, haemorrhage, retinal deetachment, intra-ocular foreign bodies and infection endophthalmitis Macular surgery involves peeling of epimacular membrane or proliferation, treating vitreomacular traction syndrome, idiopathic macular holes, retinal detachment associated with optic pit, evacuation of submacular haemorrhage and excision of choroidal neovascular membranes. Available options to treat retinal detachments are pneumatic retinopexy, scleral buckling and vitreous surgery. Proliferative vitreoretinopathy remains the important cause of failure and occurs in about 8-10% cases after retinal detachment. Vitreous surgery for ocular trauma, vitrectomy for proliferative diabetic retinopathy, macular hole surgery, submacular surgery are also discussed in detail. PMID- 11394476 TI - Revolutions in ophthalmology. AB - Cataract is the major cause of blindness worldwide. The transition of couching of cataract to most modem phaco-emulsification gained popularity with more scientific capsulorhexis. Ongoing research is taking place with laser technologies for removal of cataract. Surgery has an important role in advanced corneal diseases. Corneal transplantation is the most successful form. The refractive errors can be corrected by laser-assisted in-situ keratomileusis, intrastromal corneal rings and phacic intra-ocular lenses. Current research is going on to identify the gene responsible for glaucoma and once this is identified, medical therapy can be developed to control the disease. Thermocautery or photocoagulation and indenting the globe so as to bring the choroid nearer the break by scleral buckling forms the basis of modern surgery of retinal detachment. Vitreoretinal diseases, macular degeneration, diabetic retinopathy are no longer considered problematic. We are optimistic that more clinical breakthroughs are expected in the 21st century with new advances of ophthalmic researches. PMID- 11394477 TI - Diabetic blindness: a major challenge. AB - Diabetes mellitus, once believed to a disease of the West, is becoming an endemic to modernising and urbanising population in our country. In the last 20 years, dramatic improvement has taken place in the diabetic eye care due to availability of new information and laser technology. The diabetic blindness is now preventable provided both the physicians and patients are well informed about the need and the timing for eye check-up. The present review attempts to provide an update to physicians taking care of diabetics. PMID- 11394478 TI - Training in the United Kingdom: paradise lost? PMID- 11394479 TI - Erectile dysfunction. PMID- 11394480 TI - Sildenafil citrate. PMID- 11394481 TI - Evaluation of safety and efficacy of ketoconazole 2% and zinc pyrithione 1% shampoo in patients with moderate to severe dandruff--a postmarketing study. AB - A postmarketing study was conducted on 236 patients from 23 centres suffering from moderate to severe dandruff with a combination of ketoconazole and zinc pyrithione (1%) for a duration of 4 weeks with 2 weeks further follow-up. Scoring of dandruff was done on a 0-10 scale for each of the 6 regions of scalp at each week up to 6 weeks. The results indicate that there was a consistent improvement in dandruff scores over the treatment period and a reduction of > 90% was seen for all areas of scalp individually as well as collectively as compared to baseline. The treatment also showed significant improvement in other signs and symptoms such as erythema and itching, with a highly favourable adverse event profile. The overall assessment for global improvement by investigators showed good-excellent results with high acceptability amongst the patient population for the treatment. A combination shampoo of ketoconazole (2%) and zinc pyrithione (1%) offers a safe and effective option in the treatment of dandruff. PMID- 11394482 TI - Men Make a Difference--2000 World AIDS Day Campaign. PMID- 11394483 TI - The principal components of response strength. AB - As Skinner (1938) described it, response strength is the "state of the reflex with respect to all its static properties" (p. 15), which include response rate, latency, probability, and persistence. The relations of those measures to one another was analyzed by probabilistically reinforcing, satiating, and extinguishing pigeons' key pecking in a trials paradigm. Reinforcement was scheduled according to variable-interval, variable-ratio, and fixed-interval contingencies. Principal components analysis permitted description in terms of a single latent variable, strength, and this was validated with confirmatory factor analyses. Overall response rate was an excellent predictor of this state variable. PMID- 11394484 TI - Situational descriptions of behavioral procedures: the in situ testbed. AB - We demonstrate the In Situ testbed, a system that aids in evaluating computational models of learning, including artificial neural networks. The testbed models contingencies of reinforcement rising an extension of Mechner's (1959) notational system for the description of behavioral procedures. These contingencies are input to the model under test. The model's output is displayed as cumulative records. The cumulative record can then be compared to one produced by a pigeon exposed to the same contingencies. The testbed is tried with three published models of learning. Each model is exposed to up to three reinforcement schedules (testing ends when the model does not produce acceptable cumulative records): continuous reinforcement and extinction, fixed ratio, and fixed interval. The In Sitt testbed appears to be a reliable and valid testing procedure for comparing models of learning. PMID- 11394485 TI - Effects of signaled versus unsignaled delay of reinforcement on choice. AB - Pigeons chose between 5-s and 15-s delay-of-reinforcement alternatives. The first key peck to satisfy the choice schedule began a delay timer, and food was delivered at the end of the interval. Key pecks during the delay interval were measured, but had no scheduled effect. In Experiment 1, signal conditions and choice schedules were varied across conditions. During unsignaled conditions, no stimulus change signaled the beginning of a delay interval. During differential and nondifferential signal conditions, offset of the choice stimuli and onset of a delay stimulus signaled the beginning of a delay interval. During differential signal conditions, different stimuli were correlated with the 5-s and 15-s delays, whereas the same stimulus appeared during both delay durations during nondifferential signal conditions. Pigeons showed similar, extreme levels of preference for the 5-s delay alternative during unsignaled and differentially signaled conditions. Preference levels were reliably lower with nondifferential signals. Experiment 2 assessed preference with two pairs of unsignaled delays in which the ratio of delays was held constant but the absolute duration was increased fourfold. No effect of absolute duration was found. The results highlight the importance of delayed primary reinforcement effects and challenge models of choice that focus solely on conditioned reinforcement. PMID- 11394486 TI - Effects of response disparity on stimulus and reinforcer control in human detection tasks. AB - In two detection experiments, university students reported whether the second of two sequentially presented tones was longer or shorter than the first by responding to stimuli presented on a touch screen. Stimulus disparity and response disparity were manipulated to compare their effects on measures of discrimination and response bias when the reinforcement ratio for correct responses was asymmetric. Choice stimuli consisted of squares filled with different pixel densities. Response disparity was manipulated by varying the difference in density between the two choice stimuli. In both experiments, decreasing stimulus disparity reduced discrimination but had no consistent effect on bias. Decreasing response disparity also reduced discrimination in both experiments, and often reduced estimates of bias. The effects of response disparity on bias were most clear in Experiment 2, in which a greater overall level of response disparity was arranged. The data show that, like corresponding research with pigeons, detection performance of human subjects can be conceptualized as discriminated operants. PMID- 11394487 TI - Reinforcer-ratio variation and its effects on rate of adaptation. AB - Six pigeons were trained in sessions that consisted of six or seven concurrent schedule components, each of which could have a different reinforcer ratio arranged in it. The components were unsignaled and occurred in a random order separated by 10-s blackouts. The overall reinforcer rate arranged in each component was 2.22 reinforcers per minute. In Experiment 1, the range of reinforcer ratios in the seven components was varied from a condition in which the ratios were always 1:1, to a condition in which the ratios varied between concurrent variable-interval 27 s extinction (EXT) and concurrent extinction variable-interval 27 s (ratios of 1:EXT, 9:1, 3:1, 1:1, 1:3, 1:9, EXT:1). In Experiment 2, the range of reinforcer ratios was always 27:1 to 1:27, and the presence and absence of the intermediate reinforcer ratios used in Experiment 1 (9:1, 3:1, 1:1, 1:3, 1:9) were investigated. Log response-allocation ratios in components changed rapidly with increasing numbers of reinforcers in components, and Experiment 1 showed that sensitivity to reinforcement was usually higher when the range of reinforcer ratios was greater. When the range of reinforcer ratios was kept constant in Experiment 2, the presence or absence of less extreme reinforcer ratios had no clear effect on sensitivity. At a local level, individual reinforcers had predictable quantitative effects on response ratios: Successive same-alternative reinforcers in a component had rapidly diminishing effects in both experiments. Reinforcers obtained on the opposite alternative to one or more prior reinforcers always had large effects on preference, and these changes were greater when the range of reinforcer ratios was greater. The effects of such reinforcers in changing preference were enhanced, and produced clear preference reversals, when intermediate reinforcer ratios were absent in Experiment 2. Two processes, one local to reinforcers and one with a longer time course, may be necessary to account for these results. PMID- 11394488 TI - Changing behavior within session: cyclicity and perseverance produced by varying the minimum ratio of a variable-ratio schedule. AB - Four pigeons repeatedly chose between a fixed-ratio (FR) 20 and a variable-ratio (VR) 40 schedule of reinforcement, in which the minimum ratio of the VR cycled within each session. The minimum ratio ascended and descended (ASCDESC), descended and ascended (DESCASC), or remained constant (unchanging). In Phase 1, 2 birds (Group 1) were exposed to ASCDESC series and 2 birds (Group 2) were exposed to the DESCASC series. Choice proportions changed with the cycling minimum ratio for Group 2 but not for Group 1. In Phase 2, Group 1 subjects were exposed to the DESCASC series and Group 2 subjects were exposed to the unchanging condition. Although Group 1's choice proportions appeared to be undifferentiated in Phase 2, Group 2's choice proportions continued to cycle for more than 100 sessions. Group 2 subjects were then moved to the ASCDESC series in the third phase, and choice proportions cycled with the minimum ratio as in the first phase. The descending portion of the series was the more powerful determinant of cyclicity. Response rates also changed with the minimum component ratio, a finding that goes against the claim of universality of a rise-and-fall within session pattern of responding. That preference varied despite the constancy of the average ratio requirement suggests nonlinear averaging in quantitatively representing a variable schedule's value. The strong perseverance observed also lends support to a growing body of literature on history effects. PMID- 11394489 TI - Effects of electroacupuncture on blood-brain barrier after cerebral ischemia reperfusion in rat. AB - The effect of electroacupuncture (EA) on the blood-brain barrier (BBB) after cerebral ischemia-reperfusion (I-R) was investigated in rats. The dye Evans Blue (EB) was used as a tracer for assessing the disruption of BBB. Fluorescence quantification of EB was performed to explore the temporal pattern of permeability of BBB after the cerebral I-R with a fluorescence spectrophotometer (HITACHI 650-60). Furthermore, the morphology of BBB opening was detected under confocal laser scanning microscopy system. It was found that the BBB opening after cerebral I-R was biphasic. A rare scattered extravasation of EB was detected 2 hours after cerebral I-R. The EB extravasation reached its first peak at 6 h then decreased at 24 h and increased again at the time-point of 48h after cerebral I-R. EA can attenuate the disruption of BBB after cerebral I-R. EA could not only limit the area of extravasation of EB, but also reduce the concentration of extravasation of EB in the rat brain after cerebral I-R. The results indicated that one of the mechanisms of curative effect of EA on the cerebral ischemia might be due to its function of protecting the integrity of BBB. PMID- 11394490 TI - Thermovisual evaluation of acupuncture points. AB - The aim of our study is to evaluate the temperature distributions around the acupuncture points and channels by visualizing the infrared emissions emanated from the human skin. Our thermal imaging system that is the most sensitive to 10um long infrared wavelength allows us to recognize the actual location of acupuncture point and analyze its functional condition by examining the temperature gradient between the point and its surrounding skin area. The temperature perception of acupuncture point on extremities is especially important since its location and functional condition have a tendency to be altered if one contract disease such as immunodeficiency, arterial hypertension, and dyskinesia of biliary ducts. Also, thermovisual examinations over the acupuncture points allow us to perform the objective medical treatments by observing the transition of temperature gradients. PMID- 11394491 TI - Transcutaneous electrical nerve stimulation (TENS): the effect of electrode placement upon cutaneous blood flow and skin temperature. AB - In the current study the effect of electrode placement on cutaneous blood flow and skin temperature were assessed using laser Doppler flowmetry. Following approval from the University's ethical committee, 30 subjects were recruited (15 male: 15 female) and randomly assigned to a control or one of two treatment groups (n=10 all groups). TENS was applied either over the median nerve or to the Hegyu acupuncture point (L.I.4) for 15 minutes. Blood flow and skin temperature data were recorded during TENS and for 15 minutes post TENS. Analysis of results showed significant differences between groups for cutaneous blood flow (p=0.0001; repeated measures ANOVA). There was a significant increase in blood flow in the TENS median nerve group compared with the other two groups during TENS application. No significant changes in skin temperature were observed between groups. This study demonstrates that the effect of TENS upon cutaneous blood flow is dependent upon electrode placement sites. PMID- 11394492 TI - Melatonin might be one possible medium of electroacupuncture anti-seizures. AB - To explore the alteration of melatonin (MT) levels in pineal, hippocampus and serum during seizure crises and electroacupuncture (EA) anti-seizures, we established a rat seizure model by microinjecting benzylpenicillin into hippocampus. EA was performed on "Fengu" (DU 16) and "Jinsuo" (DU 8) acupoints in rats. Electroencephalogram (EEG) of rats was recorded and the relative power (RP) of 1 approximately 30 HZ band of EEG was analyzed. A capillary electrophoresis electrochemical detection method was used to determine MT contents. Our results indicated that MT level was elevated in pineal and hippocampus, and first had no change then significantly evaluated in serum during seizure crisis. The elevation of MT level was greatly potentiated with 30 min EA treatment (P<0.05). Meanwhile, the degree of seizures and the increases of EEG RP induced by seizures were significantly reduced (P<0.05). Because MT was considered as an antistressor and a natural downregulator of epileptiform activity, we postulate that the elevation in MT level during seizures may be one endogenous mechanism that counteracts convulsions and seizure-induced stress. A further elevation of MT levels with EA treatment suggests that MT might be one of the possible mediums of EA anti seizures. PMID- 11394493 TI - Alteration of orphanin FQ immunoreactivity and ppOFQ mRNA by combination of melatonin with electroacupuncture. AB - The aim of the present study was to observe the alternation of central orphanin FQ (OFQ, also known as nociceptin) system while electroacupuncture (EA) combined with melatonin (MEL). The experiments were carried out to investigate the changes of OFQ-like immunoreactivity and prepro-orphanin FQ (ppOFQ) mRNA in some certain nuclei of the rat brain. Using immunohistochemical technique we found that the level of OFQ-like immunoreactivity was increased significantly in some pain modulation-related nuclei, such as ventromedial hypothalamic nucleus, raphe magnus nucleus, dorsal raphe nucleus and periaqueductal gray (PAG) after intraperitoneal (i.p.) injection of MEL 60 mg/kg, and it was further enhanced while MEL combined with EA. By using in situ hybridization, we found that ppOFQ mRNA expression was decreased in the same nuclei after the administration of MEL, and further decreased following the combination of EA and MEL. The results suggested that attenuating the release and synthesis of OFQ in the brain is one of the mechanisms that melatonin promotes acupuncture analgesia. PMID- 11394494 TI - Antinociceptive effects of bee venom acupuncture (apipuncture) in rodent animal models: a comparative study of acupoint versus non-acupoint stimulation. AB - From a clinical perspective, the alternative forms of acupoint stimulation including electroacupuncture, moxibustion and acupressure appear to have more potent analgesic effects than manual needle acupuncture. Bee venom (BV) injection has also been reported to produce persistent nociceptive stimulation and to cause neuronal activation in the spinal cord. In previous study, we observed that BV stimulation into acupoint, namely BV acupuncture or Apipuncture, produced more potent anti-inflammatory and antinociceptive potency in rodent arthritis model as comparing with that of non-acupoint injection. Based on previous report, we decided to further investigate that BV injection into an acupoint produces antinociception as a result of its potent chemical stimulatory effect in both abdominal stretch assay and formalin test. Different doses of BV were injected into an acupoint or a non-acupoint 30 min prior to intraplantar formalin injection or intraperitoneal acetic acid injection. Using the abdominal stretch assay, we found that the high dose of BV (1:100 diluted in 20microl saline) produced a potent antinociceptive effect irrespective of the site of BV injection. In contrast the antinociceptive effect observed in both the writhing and formalin tests following administration of a low dose of BV (1:1000 diluted in 20microl saline) was significantly different between acupoint and non-acupoint sites. BV injection into an acupoint (Zhongwan, Cv. 12) was found to produce significantly greater antinociception than non-acupoint injection (10 mm from Zhongwan, Cv. 12) in the abdominal stretch assay. Similarly, in the formalin test, acupoint (Zusanli, St. 36) injection of BV produced more potent antinociception than non-acupoint injection (gluteal muscle). In contrast, BV injection into an arbitrary non-acupoint site on the back did not produce antinociception in either the writhing or formalin test. These results indicate that BV injection directly into an acupoint can produce a potent antinociceptive effect and suggest that this alternative form of acupoint stimulation (Apipuncture) may be a promising method for the relief of pain. PMID- 11394496 TI - MHC class II-restricted tumor antigens recognized by CD4+ T cells: new strategies for cancer vaccine design. AB - The adoptive transfer of tumor-infiltrating lymphocytes (TIL) can mediate tumor regression in patients with melanoma. This finding has led to the identification and characterization of tumor-associated antigens recognized by CD8+ TIL. Several clinical trials based on the genes recognized by these CD8+ T cells have been attempted, but with only limited success. Meanwhile, increasing evidence has demonstrated that CD4+ T cells play important roles in generating and maintaining antitumor immune responses in animal models. These data suggest that it may be necessary to engage both CD4+ and CD8+ T cells for more effective antitumor immunotherapy. In this report, we review emerging molecular approaches in cloning major histocompatibility complex (MHC) class II restricted tumor antigens recognized by CD4+ T cells as well as approaches to identify new MHC class II restricted epitopes from known tumor antigens recognized by CD8+ cytotoxic T lymphocytes and/or antibodies. Progress made in this field has shed light on the roles of tumor antigen-specific CD4+ T cells in humans; it has also provided new insights into the understanding of tumor genesis and the interaction between tumor and the immune system. More importantly, the discovery of MHC class II restricted tumor antigens has provided opportunities for developing a new generation of cancer vaccines aimed at eliciting both CD4+ and CD8+ T-cell responses against tumor. PMID- 11394495 TI - Hierarchy, tolerance, and dominance in the antitumor T-cell response. PMID- 11394497 TI - Antitumor effect on murine renal cell carcinoma by autologous tumor vaccines genetically modified with granulocyte-macrophage colony-stimulating factor and interleukin-6 cells. AB - The authors evaluted the efficacy of vaccination with murine renal cell carcinoma (Renca) secreting the granulocyte-macrophage colony-stimulating factor (GM-CSF) gene and interleukin-6 (IL-6) gene for the treatment of Renca tumor. Murine GM CSF and murine IL-6 genes were introduced and expressed in Renca cells (Renca-GM CSF and Renca-IL-6). For a prevaccination study, wild-type Renca cells were injected subcutaneously into Balb/c mice that had been vaccinated three times with inactivated wild-type Renca, Renca-GM-CSF, Renca-IL-6, or a mixture of Renca GM-CSF and Renca-IL-6 cells 7, 14, and 21 days before this tumor inoculation. For vaccination experiments, Renca tumor-bearing (8 to 10 mm) mice were injected subcutaneously weekly for 3 weeks with inactivated wild-type Renca cells, or either one or a combination of Renca-GM-CSF and Renca-IL-6. A nonvaccinated control was included in all experiments. The animals were monitored for survival and tumor development for 8 weeks. Mice inoculated with wild-type Renca alone died from the tumor within 35 days. Renca-IL-6 grew slower than wild-type Renca (p < 0.05). No tumor was produced by Renca-GM-CSF. Prevaccination with the combination of Renca-GM-CSF and Renca-IL-6 prevented subsequently inoculated wild type Renca from forming tumors, and prevaccination with either one of them, compared with prevaccination with wild-type Renca, retarded tumor growth and prolonged survival time. Tumor-bearing mice vaccinated with wild-type Renca died within 42 days. Vaccination with Renca-GM-CSF or Renca-IL-6 alone prolonged the survival time, but only Renca-GM-CSF drastically reduced the tumor size. Vaccination with the combination of them achieved complete remission. Neither of the cytokine-secreting cells enhanced the expression of MHC class I or II molecules. Autologous tumor cell vaccine secreting GM-CSF is effective in preventing and treating established tumors. Its efficacy is enhanced by the cosecretion of IL-6. PMID- 11394498 TI - Identification of a new shared HLA-A2.1 restricted epitope from the melanoma antigen tyrosinase. AB - Tyrosinase has many advantages as a target antigen for the immunotherapy of patients with melanoma because it is expressed in nearly all melanoma specimens with a high degree of cellular homogeneity, and its distribution in normal tissues is limited to melanocytes. To broaden our ability to direct cellular immune responses against this protein, we pursued an investigation to identify new shared human leukocyte antigen (HLA)-A2.1 restricted epitopes from tyrosinase. Peptides were synthesized that fit a permissive HLA-A2.1 binding motif and did not span common sites of polymorphism. The binding affinity of each peptide to HLA-A2.1 relative to a standard peptide with intermediate binding affinity was evaluated in a competitive inhibition assay. Twelve peptides were selected that had binding affinities within 80% of that of the standard peptide, and these were used to stimulate peripheral blood mononuclear cells (PBMC) in vitro from three HLA-A2.1+ patients with metastatic melanoma. Cytotoxic T lymphocytes that specifically recognized peptide-pulsed target cells as well as HLA-A2.1+ tyrosinase+ melanoma cells were raised from one patient with tyrosinase:8-17 (CLLWSFQTSA). To evaluate further the immunogenicity of this peptide, PBMC from 23 HLA-A2.1+ patients were stimulated in vitro with tyrosinase:8-17. Eleven bulk T-cell cultures demonstrated specific peptide recognition, and six of these also recognized HLA-A2.1+ tyrosinase+ melanoma cells. These data suggest that tyrosinase:8-17 may be clinically useful for the treatment of patients with melanoma. PMID- 11394499 TI - Polyclonal antibodies against gp185HER2 peptides: their putative role in the identification of a particular HER2 status in patients with breast cancer. AB - The HER2 oncogene and its relative oncoprotein, gp185HER2, a transmembrane glycoprotein belonging to the epidermal growth factor receptor family, are overexpressed in a wide range of solid tumors including breast and ovarian cancer. In patients with breast cancer, both humoral and cell-mediated HER2 immune responses have been found as well as in some patients with gp185HER2 nonoverexpressing tumors. To establish whether peptide sequences identified as HLA-A2-restricted T-cell epitopes are expressed in breast tumor cell lines and tissues, we produced and characterized by different methodologic approaches polyclonal antibodies raised against four gp185HER2 peptides. Two of the antibodies recognized peptides eluted from the HLA-A2 groove of the mDAmB231 breast cancer cell line expressing a basal level of gp185HER2. Paraffin-embedded primary and metastatic breast tumors were specifically immunostained by all four reagents, thereby showing an overlapping reactivity. When this immunoreactivity was compared with that obtained using two different monoclonal antibodies, in 105 breast primary tumors and 36 corresponding lymph node metastases, we identified a subset of tumors that were negative with anti-gp185HER2 monoclonal antibodies and positive with the four antipeptide antibodies. Our novel observations provide in vivo evidence of the complexity involved in evaluating HER2 expression, and open a new path for understanding the biologic significance of HER2 status in breast tumors. PMID- 11394500 TI - Equipotent generation of protective antitumor immunity by various methods of dendritic cell loading with whole cell tumor antigens. AB - Multiple clinically applicable methods have been used to induce dendritic cells (DCs) to express whole cell tumor antigens, including pulsing DCs with tumor lysate, and mixing DCs with apoptotic or live tumor cells. Herein we demonstrate, using two different tumor systems, that these methods are equipotent inducers of systemic antitumor immunity. Furthermore, tumor lysate pulsed DC vaccines generate more potent antitumor immunity than immunization with irradiated tumor cells plus the classic adjuvant, Corynebacterium parvum. PMID- 11394501 TI - Avoiding tolerance against prostatic antigens with subdominant peptide epitopes. AB - A potential novel therapy for prostate cancer is the induction of immune responses to normal prostate-associated antigens (PAA). One approach is to use synthetic peptides from PAA to educate T cells as a means of developing a defined and specific immunotherapy for prostate cancer. A likely major hurdle when using normal PAA for this type of therapy is the tolerance that the immune system may already have for PAA. To evaluate mechanisms for overcoming tolerance, the authors assessed the level of tolerance to SV40T antigen in a transgenic mouse. The SV40T antigen is selectively expressed in the prostates of mice from the transgenic adenocarcinoma mouse prostate (TRAMP) model. The authors have shown that TRAMP mice are tolerant to a dominant cytotoxic T-lymphocyte (CTL) epitope from the SV40T antigen compared with nontransgenic littermates. The tolerance was exhibited as early as 4 weeks and as late as 24 weeks. The use of multiple injections of an oligonucleotide that contains an unmethylated CpG induced high levels of hematopoiesis but did not overcome the tolerance. Injection of an antibody to activate CD40 increased the CTL response in normal mice but also did not overcome tolerance. However, tolerance in the TRAMP mice was avoided when an epitope that had previously been characterized as a subdominant epitope was administered. The authors are investigating the potential of subdominant epitopes to induce prostatitis and antitumor responses. The results of this work should facilitate the development of immune-based therapies for prostate cancer. PMID- 11394502 TI - Generation of cytotoxic T lymphocytes specific for human cytomegalovirus using dendritic cells in vitro. AB - For the adoptive immunotherapy in immunodeficient bone marrow transplant recipients to prevent and treat human cytomegalovirus (HCMV)-associated diseases, HCMV-pulsed dendritic cells (DCs) were used as antigen-presenting cells for the induction of cytotoxic T lymphocytes (CTLs) specific to HCMV antigens in vitro. The antiviral CTL responses induced by HCMV-pulsed DCs were as highly efficient as those induced by HCMV-infected dermal fibroblasts, and endogenous viral gene expression was not required to induce virus-specific T-cell lines. The strong cytotoxic activity against HCMV-pp65, known as HCMV major antigen, was identified using autologous B lymphoblastoid cell line expressing pp65 antigen. The cytotoxic activity toward HCMV-infected target cells was found to be mediated primarily by CD8+ T cells, although both CD8+ cells and CD4+ cells were able to lyse autologous virus-infected target cells. The CTLs contained a mixture of effector cells that recognized virus peptides in the context of major histocompatibility complex. This system may be useful for defining the cellular immune response to HCMV and for the treatment of HCMV infection in immunocompromised patients. PMID- 11394503 TI - Meta-analysis of adjuvant immunochemotherapy using OK-432 in patients with resected non-small-cell lung cancer. AB - The benefits of immunochemotherapy with a penicillin-treated, lyophilized preparation of Streptococcus pyogenes, OK-432 (Picibanil), were reassessed in patients with resected non-small-cell lung cancer through a meta-analysis based on data from 1,520 patients enrolled in 11 randomized clinical trials. All 11 trials were started before 1991, and the subjects had been followed up for at least 5 years after surgery and randomization. In these trials, standard chemotherapy was compared with the same therapy plus OK-432. The endpoint of interest was overall survival, and analysis was based on intent-to-treat population without patient exclusion. Data were analyzed using the Mantel Haenszel method. The 5-year survival rate for all eligible patients in the 11 trials was 51.2% in the immunochemotherapy group versus 43.7% in the chemotherapy group. The odds ratio (OR) for overall survival was 0.70 (95% CI = 0.56-0.87, p = 0.0010). Analysis of four trials in which central randomization was performed also reconfirmed a significantly longer survival time for the immunochemotherapy group (OR = 0.66, 95% CI = 0.44-1.00, p = 0.049). Based on these results of meta analysis, it is postulated that postoperative adjuvant immunochemotherapy using OK-432 might improve the survival of patients after resection of non-small-cell lung cancer. PMID- 11394504 TI - Sequential administration of interferon-gamma, GM-CSF, and interleukin-2 in patients with metastatic renal cell carcinoma: results of a phase II trial. AB - Various cytokine combinations have been tested for efficacy in the treatment of metastatic renal cell carcinoma (MRCC). Because several immunologic synergisms between granulocyte-macrophage colony-stimulating-factor (GM-CSF) and interleukin 2 (IL-2) have been demonstrated, this phase II trial was conducted on the efficacy and toxicity of subcutaneous, sequentially administered, interferon gamma (IFNgamma), GM-CSF, and IL-2. Fifty-five consecutive patients with MRCC were treated with 100 microg recombinant IFNgamma1b administered thrice weekly during weeks 1 and 4, followed by 400 microg GM-CSF on 5 consecutive days during weeks 2 and 5. In weeks 3 and 6, patients received 4.5 MU recombinant IL-2 from days 1 to 4. The treatment was repeated every 8 weeks. Five (10%) of patients experienced an objective response (complete response [CR]: 2%, partial response [PR]: 8%). Fourteen (26%) patients had stable disease with a median duration of 19 months (6-47+). The median overall survival was 12 months (range: 0.3-44 months). No toxicity greater than World Health Organization grade II was observed, with fever (43%) and erythema (43%) being the most frequent side effects. Compared with other phase II trials with IFN-gamma and IL-2 alone, the addition of GM-CSF failed to improve response or survival in patients with MRCC. PMID- 11394505 TI - Tumor cell expression of CD59 is associated with resistance to CD20 serotherapy in patients with B-cell malignancies. AB - The anti-CD20 chimeric monoclonal antibody rituximab (Rituxan) is used to treat patients with various B-cell tumors, including patients with plasma cell dyscrasias who have CD20+ disease. Many patients with CD20+ disease have either primary unresponsive disease or progress after initially responding to rituximab; therefore, understanding how tumor cells are, or become, resistant to rituximab is of clinical relevance. In this report, we determined whether tumor cells express antigens that block complement-mediated lysis or antibody-dependent cell mediated cytotoxicity (ADCC) and thereby contribute to rituximab resistance. We demonstrate that expression of the complement regulator CD59 is associated with resistance to rituximab-mediated complement lysis of multiple myeloma (MM) and non-Hodgkin's lymphoma (NHL) cell lines. Moreover, neutralization of CD59 using a blocking monoclonal antibody reversed resistance to rituximab-mediated complement lysis of CD20++ CD59++ ARH-77 MM cells. In addition, we demonstrate the presence of CD59 and rituximab binding on viable tumor cells from patients with MM and Waldenstrom's macroglobulinemia with progressive disease despite rituximab therapy. Last, we also examined MM and NHL B-cell lines, as well as patient tumor cells, for the expression of other antigens that may have a role in blocking ADCC activity, such as Fas ligand (FasL), MUCI, or TRAIL. FasL, MUC1, and/or TRAIL were coexpressed with complement regulators on many of these cells. These studies therefore show that complement regulators, particularly CD59 and antigens that may block ADCC, are present on various B-cell tumors and associated with rituximab resistance in patients. A prospective, clinical study is assessing the role of these antigens in mediating rituximab resistance. PMID- 11394506 TI - CD20-directed antibody-mediated immunotherapy induces responses and facilitates hematologic recovery in patients with Waldenstrom's macroglobulinemia. AB - Waldenstrom's macroglobulinemia (WM, lymphoplasmacytic lymphoma) is a B-cell lymphoproliferative disorder in which CD20 is expressed on tumor cells from most patients. Several small studies have suggested a benefit from the anti-CD20 monoclonal antibody rituximab (Rituxan, MabThera) in patients with WM. In this retrospective study, we examined the outcome of 30 previously unreported patients with WM who received treatment with single-agent rituximab (median age 60; range 32-83 years old). The median number of prior treatments for these patients was 1 (range 0-6), and 14 patients (47%) received a nucleoside analogue before rituximab therapy. Patients received a median of 4.0 (1-11.3) infusions of rituximab (375 mg/m2). Three patients received steroids with their infusions for prophylaxis of rituximab-related infusion syndrome. Overall, treatment was well tolerated. Median immunoglobulin M (IgM) levels for all patients declined from 2,403 mg/dL (range 720-7639 mg/dL) to 1,525 mg/dL (range 177-5,063 mg/dL) after rituximab therapy (p = 0.001), with 8 of 30 (27%) and 18 of 30 (60%) patients demonstrating >50% and >25% decline in IgM, respectively. Median bone marrow lymphoplasmacytic (BM LPC) cell involvement declined from 60% (range 5-90%) to 15% (range 0-80%) for 17 patients for whom pre- and post-BM biopsies were performed (p < 0.001). Moreover, 19 of 30 (63%) and 15 of 30 (50%) patients had an increase in their hematocrit (HCT) and platelet (PLT) counts, respectively. Before rituximab therapy, 7 of 30 (23.3%) patients were either transfusion or erythropoietin dependent, whereas only 1/30 (3.3%) patients required transfusions (no erythropoietin) after rituximab. Overall responses after treatment with rituximab were as follows: 8 (27%) and 10 (33%) of the patients achieved a partial (PR) and a minor (MR) response, respectively, and an additional 9 (30%) of patients demonstrated stable disease (SD). No patients attained a complete response. The median time to treatment failure for responding (PR and MR) patients was 8.0 months (mean 8.4: range 3-20+ months), and 5.0 months (mean 6.1; range 3-12+ months) for patients with SD. These studies therefore demonstrate that rituximab is an active agent in WM. Marked increases in HCT and PLT counts were noted for most patients, including patients with WM who had MR or SD. A prospective clinical trial to more completely define the benefit of single-agent rituximab in patients with WM has been initiated by many of our centers. PMID- 11394507 TI - Compound A: does it matter? PMID- 11394508 TI - Propofol-benzodiazepine interactions: insights from a "bench to bedside" approach. PMID- 11394509 TI - Production of compound A under low-flow anesthesia is affected by type of anesthetic machine. AB - PURPOSE: The purpose was to compare the concentrations of compound A in inspired gas breathed by patients produced by different types of anesthetic machines under prolonged sevoflurane low-flow anesthesia. METHODS: The anesthetic machines tested were Excel 210 SE (Datex-Ohmeda, Louisville, CO), Cicero (Drager, Lubeck, Germany), and AS/3 ADU (Datex-Ohmeda, Louisville, CO). Anesthesia expected to last more than four hours was maintained with 2.0% sevoflurane and nitrous oxide (0.5 L x min(-1))/oxygen (0.5 L x min(-1)). The concentrations of compound A, obtained from the inspiratory limb of the circle system, were measured using a gas chromatograph. RESULTS: When Excel and Cicero were used, concentrations of compound A increased steadily from the baseline values to 28 and 29 (mean) ppm, respectively, at two hours after exposure to sevoflurane and became constant. There was no significant difference between the concentrations of compound A produced by these anesthetic machines. In contrast, the new anesthetic machine AS/3 was associated with lower concentrations of compound A (6 ppm at one hour, P <0.05 compared with Excel and Cicero), and the concentration did not change significantly thereafter. CONCLUSION: In spite of the use of a conventional carbon dioxide (CO2) absorbent with strong bases, the anesthetic machine AS/3 with a small volume of canister/soda lime (900 ml/700 ml) produced lower concentrations of compound A than those produced by the other machines. PMID- 11394510 TI - Midazolam premedication reduces propofol dose requirements for multiple anesthetic endpoints. AB - PURPOSE: This study investigates the interactions between midazolam premedication and propofol infusion induction of anesthesia for multiple anesthetic endpoints including: loss of verbal contact (LVC; hypnotic), dropping an infusion flex (DF; motor), loss of reaction to painful stimulation (LRP; antinociceptive) and attainment of electroencephalographic burst suppression (BUR; EEG). METHODS: In a double blind, controlled, randomized and prospective study, 24 ASA I-II patients received either midazolam 0.05 mg x kg(-1) (PM; n = 13) or saline placebo (PO; n = 11) i.v. as premedication. Twenty minutes later, anesthesia was induced by propofol infusion at 30 mg x kg(-1) x hr(-1). ED50, ED95 and group medians for times and doses were determined and compared at multiple anesthetic endpoints. RESULTS: At the hypnotic, motor and EEG endpoints, midazolam premedication significantly and similarly reduced propofol ED50 (reduction: 18%, 13% and 20% respectively; P <0.05 vs unpremedicated patients) and ED95 (reduction: 20%, 11% and 20% respectively; P <0.05 vs unpremedicated patients). For antinociception (LRP), dose reduction by premedication was greater for propofol ED95 (reduction: 41%; P <0.05 vs unpremedicated patients) than ED50 (reduction: 18%; P <0.05 vs unpremedicated patients). Hemodynamic values were similar in both groups at the various endpoints. CONCLUSIONS: Midazolam premedication 20 min prior to induction of anesthesia reduces the propofol doses necessary to attain the multiple anesthetic endpoints studied without affecting hemodynamics in this otherwise healthy population. The interaction differs for different anesthetic endpoints (e.g., antinociception vs hypnosis) and propofol doses (e.g., ED50 vs ED95). PMID- 11394511 TI - Perioperative medical management and outcome following thymectomy for myasthenia gravis. AB - PURPOSE: To describe the evolution of the perioperative management of myasthenia gravis (MG) patients undergoing thymectomy and to question the need for systematic postoperative ventilation. CLINICAL FEATURES: We collected data retrospectively from 36 consecutive MG patients who underwent thymectomy over a 21-yr period, via transthoracic, -cervical or -sternal incisions (n=5, n=7, n=24, respectively). From 1980 to 1993, a balanced anesthetic technique (n=24) included various inhalational agents with opiates and myorelaxants (in eight cases); 22 patients were admitted to the intensive care unit (ICU). Since 1994, i.v. propofol was combined with epidural bupivacaine and sufentanil (n=12); all patients were admitted to the postanesthesia care unit. Short-term postoperative ventilation (median time four hours, range from three to 48 hr) was required in eight patients who had longer hospital stay (median stay=12 days, range (8-28) vs five days (4-15) for patients with early extubation, P <0.05) but similar clinical improvement six months after thymectomy. Postoperative ventilatory support was required more frequently when a balanced anesthetic technique was used (odds ratio=4.2 (1.1-9.7), P=0.03) and particularly when myorelaxants were given (odds ratio=13.9 (2.1-89.8), P=0.009). Leventhal's scoring system had low sensitivity (22.2%) and positive predictive values (25%). CONCLUSIONS: Our data show that the severity of MG failed to predict the need for postoperative ventilation. A combined anesthetic technique was a safe and cost-effective alternative to balanced anesthesia as it provided optimal operating conditions and resulted in fewer admissions in ICU and shorter hospital stays. PMID- 11394512 TI - An inventory of Canadian anesthesiology. Human research from 1995 through 1999. AB - PURPOSE: The year 2000 provides a symbolic opportunity to assess the past initiatives in anesthesia research. As in many other fields, medical research has benefited from utilizing computerized data bases to facilitate enumerating areas of interest. We have created a baseline survey of past research in the fields of anesthesia, anesthetics, analgesia, and analgesics to highlight Canadian studies. METHODS: The survey was undertaken using the Medical Literature Analysis and Retrieval System (MEDLARS) medical literature archive for the years 1995 through 1999. The principal categories and sub-categories of MEDLARS' anesthesia classifications were counted for 70 countries contributing to the archive. RESULTS: Canadian contributions ranged from 141 (1992) to 185 (1999) and represented annually 3% of the world total in the anesthesia categories. The greatest number of studies (30-38%) were about adults aged 19 to 44 yr, and there were between 4% and 14% more studies of females than males. "Pharmacology" and "therapeutic use" were the most frequent topics, lidocaine, fentanyl, and propofol were the most studied anesthetics, and non-steroidal anti inflammatories, opium, morphine, and fentanyl were the most studied analgesics. Among the types of studies, those classified as "quality of health care" occurred most frequently (16%). Canadian trends closely follow world trends. CONCLUSION: The collected counts provide a comprehensive overview of research trends for the past five years. PMID- 11394513 TI - Hypnotic and cardiovascular effects of proprietary and generic propofol formulations do not differ. AB - PURPOSE: The aim of the study was to compare the potency of two different propofol formulations: proprietary and generic propofol using the bispectral index (BIS) monitoring. METHODS: Forty female patients undergoing breast surgery received propofol 3 mg x kg(-1) followed by propofol infusion adjusted to maintain a 40% BIS value, supplemented by 50% nitrous oxide. Proprietary or generic propofol was administered in a randomized double-blind manner. RESULTS: The propofol mg/BIS% ratio obtained after the bolus dose, the cumulative infused propofol mg/BIS% ratio at the end of each five-minute interval and the total dose of propofol administered as bolus + infusion were similar between the two groups. The two groups did not differ with regard to systolic and diastolic blood pressure, heart rate, end-tidal carbon dioxide and arterial oxygen saturation. CONCLUSION: The two propofol formulations are equipotent when hypnotic effect is assessed by BIS monitoring. PMID- 11394514 TI - Increased noradrenaline release from rat preoptic area during and after sevoflurane and isoflurane anesthesia. AB - PURPOSE: To study the effects of sevoflurane and isoflurane on noradrenaline release from the rat preoptic area (POA). METHOD: Sixteen male Wistar rats were studied. A microdialysis probe with a 2 mm long semipermeable membrane was implanted in the POA. Dialysates were collected at intervals often minutes. After obtaining five control samples for 50 min, 30 min inhalation of 3% sevoflurane or 1.8% isoflurane was performed. After cessation of the inhalation, five more samples were obtained for 50 min as recovery phase. Noradrenaline (NA) concentration in the dialysates was measured by high pressure liquid chromatography with an electrochemical detector. RESULTS: Both sevoflurane and isoflurane caused marked increases in NA release from the rat POA (sevoflurane 233% at 20 min, isoflurane 357% at ten minutes after the start of inhalation). The marked NA releases were also observed during the emergence from sevoflurane and isoflurane anesthesia (sevoflurane 269% at 20 min, isoflurane 368% at ten minutes in the recovery phase). CONCLUSION: This study suggests that enhanced release of NA in the POA during sevoflurane and isoflurane may explain the excitatory phase observed during the peri-anesthetic period with these agents. PMID- 11394515 TI - Intrathecal fentanyl does not modify the duration of spinal procaine block. AB - PURPOSE: To document the clinical characteristics of spinal procaine with or without the addition of fentanyl in light of the failure rate observed previously with procaine 10%. METHODS: In a randomized, prospective, double-blind study, 52 patients received spinal anesthesia with 100 mg procaine and either saline 0.9% (0.4 ml) (CONTROL group) or 20 microg fentanyl (0.4 ml) (FENTANYL group). Sensory anesthesia to needle prick was evaluated each minute for ten minutes, every three minutes for 33 minutes and every five minutes until regression to T10. Motor block was assessed with the Bromage scale. Patients were questioned by telephone for pain suggesting transient radicular irritation (TRI) 48 hr later. RESULTS: Mean time to reach highest sensory level, maximum number of segments blocked and mean time for regression of the sensory level to T10 showed no difference. Time to recuperate to full flexion of knees and feet (Bromage 4) showed no difference. Nine patients had nausea (five in CONTROL group and four in FENTANYL group) and nine had pruritus (three in CONTROL group and six in FENTANYL group). No patient reported pain suggesting TRI. CONCLUSION: Spinal procaine is appropriate for short-duration surgery. Fentanyl does not change the characteristics of the block or the incidence of side effects associated with spinal procaine. PMID- 11394516 TI - Spinal anesthesia: a comparison of procaine and lidocaine. AB - PURPOSE: To compare spinal procaine to spinal lidocaine with regard to their main clinical characteristics and incidence of transient radicular irritation (TRI). METHODS: In this randomized, double-blind, prospective study, patients (two groups, n=30 each) received either 100 mg of lidocaine 5% in 7.5% glucose (Group L) or 100 mg of procaine 10% diluted with 1 ml cerebrospinal fluid (Group P). After spinal anesthesia, segmental level of sensory block was assessed by pinprick. Blood pressure and the height of the block were noted each minute for the first ten minutes, then every three minutes for the next 35 min and finally every five minutes until regression of the block to L4. Motor blockade was evaluated using the Bromage scale. To evaluate the presence of TRI, each patient was questioned 48 hr after surgery. RESULTS: Time to highest sensory level and to maximum number of segments blocked showed no difference between groups. Mean time for sensory regression to T10 and for regression of the motor block were shorter in Group P. Eighty minutes following injection, sensory levels were lower in Group P. Five patients had inadequate surgical anesthesia in Group P and only one in Group L. No patient in Group P had TRI (95% CI 10-12%) while eight (27%) in Group L did (95% CI 12-46%). CONCLUSIONS: Procaine 10% was associated with a clinical failure rate of 14.2%. This characteristic must be balanced against an absence of TRI, which occurs more frequently with the use of lidocaine 5%. PMID- 11394517 TI - Cesarean section in a patient with syringomyelia. AB - PURPOSE: To describe the anesthetic management of Cesarean section in a patient with syringomyelia. CLINICAL FEATURES: A 27-yr-old pregnant woman with syringomyelia was scheduled to undergo elective Cesarean section. At the age of 25 yr, she had begun to experience headaches, and at the age of 26 yr, a diagnosis of syringomyelia of the upper spinal cord was made on the basis of magnetic resonance imaging findings. No symptoms other than headache were noted preoperatively. General anesthesia was used for the Cesarean section. After the administration of 1 mg vecuronium as a priming dose, 5 mg vecuronium were injected. At the onset of clinical muscle weakness, 225 mg thiamylal were promptly administered as the induction agent and the patient was intubated (timing principle with priming method) and pressure on the cricoid cartilage applied to prevent regurgitation of stomach contents. Anesthesia was maintained with oxygen, nitrous oxide and isoflurane at a low concentration. Mild hyperventilation was used throughout the procedure. Anesthesia and surgery proceeded without any problem, response to vecuronium was clinically normal and recovery was uneventful. Neurological status remained normal. CONCLUSION: We report the safe use of general anesthesia for Cesarean section in a patient with syringomyelia. Precautions were taken to avoid increases in intracranial pressure and our patient experienced no untoward neurologic event. PMID- 11394518 TI - Separation of omphalopagus conjoined twins using combined caudal epidural-general anesthesia. AB - PURPOSE: To describe the anesthetic management of newborn omphalopagus conjoined twins undergoing a series of diagnostic and surgical procedures which culminated in successful separation at one month of age. CLINICAL FEATURES: Evaluations of the extent of shared organ systems were carried out without the need for anesthesia. The twins were anesthetized twice, once for insertion of skin expanders, and later for surgical separation. Various airway management techniques were utilized. To facilitate surgical separation, caudal epidural catheters were inserted in an effort to provide both operative and post-operative analgesia. In addition, prior to the induction of anesthesia, the extent of cross circulation between twins was assessed. CONCLUSION: Caudal epidural catheters can be used to provide both operative and post-operative analgesia. Early extubation, another benefit of regional analgesia, was not achieved because both twins developed respiratory failure in the immediate postoperative period. Testing for the extent of cross-circulation between twins proved valuable, allowing for detailed scripting of the complex induction sequence and airway management. PMID- 11394519 TI - Poor man's LMA: achieving adequate ventilation with a poor mask seal. AB - PURPOSE: This report describes a technique of ventilation prior to laryngoscopy and intubation that proved to be simple, inexpensive and effective for a patient whose airway evaluation suggested difficult mask ventilation. The technique is called Poor Man's LMA. CLINICAL FEATURES: A 60-yr-old male, measuring 170 cm, weighing 117 kg, edentulous and with a full beard, was to undergo uvulopalatopharyngoplasty. After induction of general anesthesia with a hypnotic, analgesic and non-depolarizing muscle relaxant, it was soon determined that mask bag ventilation was difficult due to an inadequate seal between the mask and the patient's full beard. To improve ventilation, an endotracheal tube was placed into the oropharynx, the lips and nose compressed by a colleague in order to prevent gas egress, and effective manual ventilation established by connecting the circle system to the endotracheal tube. Subsequent direct laryngoscopy and intubation were accomplished without incident. CONCLUSION: The patient's clinical features made conventional mask bag ventilation difficult and inadequate. The Poor Man's LMA technique improved oxygenation and ventilation in preparation for intubation. Further investigations on the usefulness of this technique are warranted. PMID- 11394520 TI - Combined phosphodiesterase inhibition and beta-blockade in the GI104313, decreases ischemia-induced arrhythmias in the rat. AB - PURPOSE: The purpose of the present study was to evaluate the effects of GI104313, a chimeric molecule containing a phosphodiesterase inhibiting pyradazinone and a blocking phenoxpropanolamine, on ischemia-induced arrhythmias in anesthetized rats. METHOD: The coronary artery was occluded 15 min after commencing drug administration and myocardial ischemia was maintained for 30 min during which the heart rate and mean blood pressure were recorded. Cyclic AMP and GMP were determined by radio-immunoassay. RESULTS: GI104313 (0.1 micromol x kg( 1) plus 0.01 micromol x kg(-1) x min(-1) or 1 micromol x kg(-1) plus 0.1 micromol x kg(-1) x min(-1)) decreased the incidence of ventricular tachycardia (86% and 75%), ventricular fibrillation (28%, P <0.01 and 12%, P <0.001) and premature ventricular beats (164 +/- 27.0 and 114 +/- 28.5, P <0.05) following coronary artery ligation, resulting in a decrease in mortality (29% and 12%, P <0.05). Changes in cyclic nucleotide concentrations have been implicated in the genesis of ischemia-induced arrhythmias. However, in the present study GI104313 did not change the concentrations of adenosine 3':5'-cyclic monophosphate (cyclic AMP) (1.0 +/- 0.07 pmol x mg(-1), 1.0 +/- 0.05 pmol x mg(-1)) or guanosine 3':5' cyclic monophosphate (cyclic GMP) (0.025 +/- 0.008 pmol x mg(-1) protein, 0.017 +/- 0.004 pmol x mg(-1) protein) in the left ventricle during ischemia-induced arrhythmias in anesthetized rats compared to saline (0.9 +/- 0.1 pmol x mg(-1) and 0.013 +/- 0.002 pmol x m(-1), respectively). CONCLUSION: Our results demonstrate that, in rats, GI104313 induced a decrease in both incidence of arrhythmias and mortality which was not associated with changes in ventricular cyclic nucleotide content. PMID- 11394521 TI - Life-threatening pneumothorax of the ventilated lung during thoracoscopic pleurectomy. AB - PURPOSE: To report the case of a patient who underwent right thoracoscopic pleurectomy with lung exclusion and developed contralateral (left) pneumothorax with resulting life-threatening alteration of the respiratory and cardiovascular functions. CLINICAL FEATURES: A 28-yr-old male was admitted to the intensive care unit for a well tolerated, second episode of spontaneous right pneumothorax and scheduled for right thoracoscopic pleurectomy. Anesthesia was induced and maintained with sufentanil and propofol. A double lumen endotracheal tube (ETT) was inserted, its correct positioning checked clinically and by fiberoptic bronchoscopy and the patient was placed in the left decubitus position. Approximately one hour into the procedure, during the second period of right pulmonary exclusion, SpO2 values decreased within two minutes to 78%. End tidal capnography (EtCO2) values decreased to 6-8 mmHg within seconds and peak airway pressure increased to values between 50 and 60 cm H2O. Severe cyanosis, sinus bradycardia and arterial hypotension developed. The surgical procedure was stopped, propofol administration discontinued, bipulmonary ventilation reinstituted and the patient placed in the supine position which restored hemodynamic and respiratory function. Inspection and auscultation were consistent with tension left pneumothorax which was evacuated. CONCLUSION: Pneumothorax of the ventilated lung during one lung ventilation for thoracoscopic procedures must be diagnosed quickly. Reinstitution of bipulmonary ventilation should probably be the first therapeutic attitude. PMID- 11394522 TI - Spatially resolved spectroscopy (NIRO-300) does not agree with jugular bulb oxygen saturation in patients undergoing warm bypass surgery. AB - PURPOSE: Near infrared spectroscopy (NIRS) is a promising non-invasive method for continuous monitoring of cerebral oxygenation during cardiac surgery with cardiopulmonary bypass (CPB). This study was designed to study the agreement between tissue oxygen index (TOI) measured by spatially resolved spectroscopy (NIRO-300) and jugular bulb oxygen saturation (SjO2) in patients undergoing warm coronary bypass surgery. METHODS: Seventeen patients undergoing warm coronary artery bypass surgery were studied. NIRS was continuously monitored and was averaged before CPB, five, 20, 40, 60 min on CPB, five minutes before end of CPB and ten minutes after CPB to coincide with SjO2 measurements. Bypass temperature was maintained at 34-37 degrees C. RESULTS: Bland and Altman analysis showed a bias (TOI-SjO2) of -6.7%, and wide limits of agreement (from 16% to -28%) between the two methods. In addition, mean TOI was lower than mean SjO2 during and after CPB. We observed a statistically significant correlation between arterial carbon dioxide and SjO2 measurements (r2=0.33; P=0.0003), but the former did not correlate with TOI values (r2=0.001; P=0.7). CONCLUSION: Our results demonstrate a lack of agreement between SjO2 and TOI for monitoring cerebral oxygenation during cardiac surgery. We conclude that the two methods are not interchangeable. PMID- 11394523 TI - Multiple organ dysfunction syndrome: a narrative review. AB - PURPOSE: To review multiple organ dysfunction syndrome with respect to: 1) clinical measurement systems; 2) molecular mechanisms; and 3) therapeutic directions based upon molecular mechanisms. METHODS: The Medline, Cochrane, and Best Evidence databases (1996 to 2000), conference proceedings, bibliographies of review articles were searched for relevant articles. Key index words were multiple organ failure, multiple system organ dysfunction, sepsis, septic shock, shock, systemic inflammatory response syndrome. Outcomes prospectively defined were death and physiological reversal of end organ failure. RESULTS: Multiple organ dysfunction/failure (MODS) is the most common cause for death in intensive care units. The recognition of this syndrome in the last 30 yr may be due to advances in early resuscitation unmasking these delayed sequelae in those that would have died previously. Multiple organ dysfunction occurs after shock of varied etiologies and may be the result of unbridled systemic inflammation. As yet, therapy directed to prevent or improve MODS has not dramatically altered outcomes. CONCLUSION: Multiple organ dysfunction may serve as useful measure of disease severity for risk adjustment and outcome marker for quality of care and therapy provided. Anesthesiologists treating shock patients will note the subsequent development of MODS in the critical care unit and may be required to provide anesthetic support to these patients. PMID- 11394524 TI - New media. PMID- 11394525 TI - Use of the laryngeal mask in a patient with pemphigus vulgaris. PMID- 11394526 TI - Local anesthetic toxicity and milrinone. PMID- 11394527 TI - The "toothpaste method" for anesthesia of the difficult airway. PMID- 11394528 TI - Characterization of monoclonal antibodies specific for feline serum amyloid (SAA) protein. AB - Serum amyloid A (SAA) has been characterized as an inflammatory marker in many species. In this study, we have developed and characterized monoclonal antibodies (MAbs) against feline SAA (fSAA) derived from culture hybridomas. These hybridomas were produced from the fusion of Balb/c-derived myeloma s/p20-Ag14 and splenocytes from mice immunized with purified recombinant feline SAA (rfSAA). Six hybridomas secreting MAbs, M2, M5, M7, M8, M13, and M15, were selected and subcloned on the basis of their specificity to rfSAA by enzyme-linked immunoabsorbent assay (ELISA), and confirmed based on their specificity to rf-SAA by immunoblot analysis. Out of six clones, two clones (M5 and M7) showed higher reactivity with rf-SAA, and were selected for further analysis of ELISA additivity and Western blot cross-reactivity tests. As a result, M5 and M7 clones recognized the same or excessively near epitopes on rfSAA and reacted with rfSAA, fSAA and equine recombinant SAA, but showed no reaction with human recombinant SAA. Because of their specificity, these MAbs may be usefully applied in studying the measurement of SAA concentration in cat serum. PMID- 11394529 TI - Monoclonal antibody to rat galanin: production, characterization, and in vivo immunoneutralization activity. AB - A monoclonal antibody (MAb) to galanin was prepared by cell fusion of myeloma Fox NY and spleen cells from Robertsonian mice immunized with rat galanin. Hybridomas producing high-affinity antibodies were cloned in pristine-primed Balb/c mice. The antibody was purified by affinity chromatography and concentrated to 12 mg IgG/mL by dialysis. Immunoreactivity of the antibody was screened by radioimmunoassay. Ascites fluid contained approximately 10 mg/mL IgG that belong to the subclass of IgG2a as determined by enzyme-linked immunoadsorbent assay (ELISA). The titer of this IgG2a antibody entitled #G65G was 1:10,000 and the ID50 for rat galanin was 1000 fmol/mL as determined by liquid phase radioimmunoassay. Immunohistochemistry showed that this galanin MAb stains densed, beaded processes distributed to the enteric plexuses, where they appear to encircle neuronal cell bodies, to the muscle layer, where they are particularly abundant in the circular muscle layer and in the deep muscular layer, and to the mucosa. In vivo capacity of immunoneutralization by this antibody was tested in male Sprague-Dawley rats fasted for 24 h and anesthetized with urethane. Systemic injection of protein A purified galanin antibody (6 mg/rat) decreased by 70% of the inhibitory effect of intravenous galanin (2 nmol/kg/h i.v.) on gastric acid secretion induced by intracisternal TRH analog. These results show that galanin antibody #G65G is useful for in vivo immunoneutralization of galanin effects and is a valuable tool for immunohistochemical localization of galanin in gastrointestinal tissues. PMID- 11394530 TI - Characterization of a monoclonal antibody against neopterin using an enzyme linked immunosorbent assay with penicillinase as label. AB - An active ester derivative of neopterin was prepared using 4-(N-maleimidomethyl) cyclohexan 4-carboxilic acid N-hydroxy succinimide ester (MCH-NHS), conjugated to bovine serum albumin (BSA) and injected for antibody production (for both monoclonal and polyclonal antibodies). High titer antibody producing spleen cells were removed and fused with myeloma cells of Sp2/0 origin. Neopterin was conjugated to the enzyme penicillinase by a one-step glutaraldehyde method, which was used as tracer. A novel enzyme immunoassay was developed using this conjugate to screen and characterize the monoclonal antibody (MAb) produced in these experiments. After limiting dilutions, it was found that antibody produced by one clone with a Ka value of 7.6 x 10-7 mol/L was specific for a number of structurally related molecules. This clone was found to be of IgG class and IgG2a subclass. The standard curvewas constructed with a sensitivity of 10 pg/well (100 pg/mL) covering up to 1 ng/mL. PMID- 11394531 TI - Immunological characterization of a mucin-associated protein from hamster tracheal epithelial cell culture. AB - Airway mucins are high molecular mass (>10(6) dalton) glycoproteins with various types of associated molecules including glycoproteins, lipoproteins, and lipids. The study of mucin-associated proteins is limited largely due to the lack of specific probes. In this study, we produced a monoclonal antibody, MAbHT10, against a 190-kDa mucin associated-protein by immunizing mice with hamster airway mucin purified in nondissociative condition. Using HT10, the 190-kDa mucin associated protein was characterized immunologically. The 190-kDa mucin associated protein is glycoprotein and HT10 recognized carbohydrate containing portion of the protein. The association of 190-kDa protein with mucin is strong enough that heat and detergent treatment is required to dissociate it from mucin as evidenced by gel filtration chromatography, Western blot, enzyme-linked immunoadsorbent assay (ELISA), and co-immunoprecipitation. The expression of the 190-kDa protein is increased with the development of hamster tracheal epithelial cells in culture, but showed differences with the pattern of the regulation of mucin expression. Adenosine triphosphate (ATP), a known strong mucin secretagogue, dose-dependently increased mucin release but caused only marginal increase in the release of the 190-kDa protein. The MAb should be useful in the structural and functional analysis of the 190-kDa mucin-associated proteins in physiological and pathological situations such as chronic airway diseases. PMID- 11394532 TI - Phase I clinical evaluation of a neutralizing monoclonal antibody against epidermal growth factor receptor in advanced brain tumor patients: preliminary study. AB - High levels of growth factors and their receptors have been demonstrated in human tumors. Gliomas and meningiomas are characterized by overexpression of epidermal growth factor receptor (EGF-R). Ior egf/r3, is a neutralizing murine monoclonal antibody (MAb) against EGF-R, and was generated at the Cuban Institute of Oncology. The antibody recognizes EGF-R with high affinity, inhibiting tyrosine kinase activation. A clinical trial was conducted in brain tumor patients to evaluate toxicity, immunogenicity, and clinical benefit of escalating doses of the antibody. Nine patients with histologically confirmed gliomas or meningiomas, who had active or recurrent disease after receiving conventional treatment, received four intravenous doses of ior egf/r3. Total dosages ranged from 160 to 480 mg. As inclusion criteria, radioimmunoscintigraphy with the same MAb labeled with 99mTechnetium (99mTc) was performed. Immune response against the murine antibody was also evaluated. After four doses of ior egf/r3 MAb, no significant toxicity was found, except in one patient who developed a grade 4 allergic adverse event. This reaction was probably related with previous sensitization to the same MAb and the development of human anti-mouse antibodies (HAMA) response. Despite no major objective antitumor responses, eight patients had stable disease on the 6-month evaluation, and two patients remain alive after four years of MAb therapy. PMID- 11394533 TI - Differential expression and regulation of CD6 on T-cell subsets revealed by monoclonal antibody (MAb) CH11. AB - A monoclonal antibody (MAb), CH11, was developed by immunizing mice with CD4+ gammadelta T-cell receptor (TCR)+ cells. It recognized an antigen expressed in the surface membrane of T-cell lines, but not of U937, lymphoblastoid B cells (LBC), K562, Raji or Daudi cells, indicating selectivity for the T-cell lineage. In addition, it labelled 70-80% of normal peripheral blood mononuclear cells (PBMC), with high expression on the erythrocyte rosetting (E+) fraction, and low/absent expression on E- cells. However, CD4+ T cells expressed higher levels of reactivity than CD8+ or gammadelta+ T-cell receptor (TCR)+ lymphocytes in PB. Furthermore, in 7 of 10 individuals tested, 7.34+/-3.88% of unselected PBMC were CH11- CD3+ and were relatively enriched in CD8+ and in gammadelta TCR+-cells. In addition, thymic gammadelta T cells, and gammadelta lymphoproliferations from two patients were nonreactive or weakly reactive with the MAb. Activation of E+ cells with phorbol-12-myristate-13-acetate (PMA) enhanced CH11 expression uniformly, whereas activation with phytohemagglutinin (PHA) selectively down-regulated expression of the antigen on the CD8+ subset. In Western blots performed in nonreducing (NR) conditions, MAb CH11 detected a 100 kDa molecule in PBMC and Jurkat T-cell lysates. Preincubation of T cells with MAb CH11 specifically abrogated their subsequent reactivity with MAb to CD6, suggesting that MAb CH11 is recognizing an epitope of CD6. Given its function as a receptor for ligands on thymic epithelium, activated leukocytes and synoviocytes, this newly defined heterogeneity of expression and regulation of the CD6 molecule on subsets of T cells may help determine their functional repertoire in vivo. PMID- 11394534 TI - Characterization of monoclonal antibodies with specificity for the core oligosaccharide of Shigella lipopolysaccharide. AB - Monoclonal antibodies (MAbs) were prepared against different strains of Shigella, following immunization of BALB/c mice with a heat-killed preparation of Shigella. Antibody-producing hybridomas were screened in an indirect enzyme-linked immunoadsorbent assay (ELISA) and epitope specificity determined using chemically defined lipopolysaccharide, lipid, and KDO fragments. Five MAbs were characterized and the following specificities identified: 2C32E6 and 4D64B9 (reactive to S. flexneri and S. boydii), 5E45D8 (reactive with S. flexneri), 4B33D10 and 1B52F10 (all species of Shigella). The properties of 1B52F10 revealed its potential importance in immunological detection of Shigella from unknown samples, as it was able to bind to all strains of Shigella. PMID- 11394535 TI - Characterization of four new monoclonal antibodies that recognize mouse natural killer activation receptors. AB - With the aim of identifying natural killer (NK) activation receptors, we immunized BALB/c mice with (BALB/cxB6)F1 NK LAK cells and made B-cell hybridomas. These were screened for monoclonal antibody (MAb) reacting with an NK activation receptor by using an antibody-induced redirected lysis (AIRL) assay against FcR bearing P815 targets. Four hybridomas, clones 1C10, 1F10, 2D10 and 4G4, were selected for further characterization. Protein G-purified MAbs from these clones activated both resting and IL-2 activated B6 or F1 NK cells in the AIRL assay. 1F10 MAb, but not the other three MAbs, could compete for the binding of anti NK1.1 (PK136) MAb to F1 NK cells. The four MAbs were screened for their ability to bind to or activate NK cells from the mouse strains SJL/J, DBA/2, 129/J, C3H/J, and BALB.K. None showed activity except IC10, which could bind to and activate SJL/J NK cells. When members of the NKR-P1 family from both B6 mice (A, B, and C genes expressed) and SJL mice (only A and B genes expressed) were expressed in Jurkat cells and tested for their antibody reactivity, PK136 MAb was found to recognize B6 NKR-P1C and SJL/J NKR-P1B; IC10 MAb was found to recognize NKR-P1-A, -B and -C from B6, but not NKR-P1A or -B from SJL/J; and 1F10 MAb was found to react only with B6 NKR-P1C. PMID- 11394536 TI - Academic radiology..."red in tooth and claw"? PMID- 11394537 TI - Digital mammography: observer performance study of the effects of pixel size on the characterization of malignant and benign microcalcifications. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate the effects of pixel size on the characterization of mammographic microcalcifications by radiologists. MATERIALS AND METHODS: Two-view mammograms of 112 microcalcification clusters were digitized with a laser scanner at a pixel size of 35 microm. Images with pixel sizes of 70, 105, and 140 microm were derived from the 35-microm-pixel size images by averaging neighboring pixels. The malignancy or benignity of the microcalcifications had been determined with findings at biopsy or 2-year follow-up. Region-of-interest images containing the microcalcifications were printed with a laser imager. Seven radiologists participated in a receiver operating characteristic (ROC) study to estimate the likelihood of malignancy. The classification accuracy was quantified with the area under the ROC curve (Az). The statistical significance of the differences in the Az values for different pixel sizes was estimated with the Dorfman-Berbaum Metz method and the Student paired t test. The variance components were analyzed with a bootstrap method. RESULTS: The higher-resolution images did not result in better classification; the average Az with a pixel size of 35 microm was lower than that with pixel sizes of 70 and 105 microm. The differences in Az between different pixel sizes did not achieve statistical significance. CONCLUSION: Pixel sizes in the range studied do not have a strong effect on radiologists' accuracy in the characterization of microcalcifications. The low specificity of the image features of microcalcifications and the large interobserver and intraobserver variabilities may have prevented small advantages in image resolution from being observed. PMID- 11394538 TI - Percutaneous core biopsy of the breast: correlates of anxiety. AB - RATIONALE AND OBJECTIVES: The authors performed this study to investigate the level of patient anxiety immediately preceding breast biopsy and examined potential clinical and demographic correlates of anxiety. MATERIALS AND METHODS: The authors evaluated 102 women who were referred to a radiology breast clinic to undergo breast biopsy. The women were assessed with a self-report of demographic and medical items and the State Trait Anxiety Inventory (STAI) immediately before their biopsy. The STAI also was administered at 1 and 5 days after biopsy. RESULTS: The participants' mean state anxiety T score as measured with the State Trait Anxiety Inventory was 71.1 (standard deviation, 7.2). Multiple regression analysis was performed to determine the correlates of state anxiety. The variables that showed the strongest correlation with state anxiety were trait anxiety, being concerned about the results of biopsy, education (less education was associated with more anxiety), age (an older age was associated with more anxiety), and number of relatives with breast cancer. Given the expected overlap (r = 0.55) between state and trait anxiety, a second regression analysis was performed that controlled for trait anxiety. The results of this analysis also identified age, being concerned about the results of the biopsy. and number of relatives with breast cancer as relevant correlates of state anxiety. CONCLUSION: Overall, the results give some indication of the characteristics of women likely to be most anxious before biopsy. Future research should assess the effectiveness of different strategies for addressing situational anxiety. PMID- 11394539 TI - Suprarenal intraarterial infusion of alloxan and streptozotocin during balloon occlusion of the juxtarenal abdominal aorta: a simple technique for inducing diabetes mellitus in canines with reduced mortality. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate the mortality and morbidity associated with a simple technique for inducing diabetes in dogs--suprarenal intraarterial infusion of alloxan and streptozotocin during balloon occlusion of the juxtarenal abdominal aorta. MATERIALS AND METHODS: The authors attempted to induce diabetes in six purpose-bred dogs. After the dogs were fasted for 12 hours, the abdominal aorta at the level of the origin of the renal arteries was occluded with an angioplasty balloon introduced by means of a femoral approach. A 3-F microcatheter (n = 1) or infusion wire (n = 5) was introduced via the percutaneous transluminal angioplasty catheter and positioned at the level of the celiac axis, and a mixture of streptozotocin (20-25 mg/kg) and alloxan (20-25 mg/kg) was infused. Diabetes was considered to have been induced if the dogs experienced sustained hyperglycemia. RESULTS: There were no deaths during the follow-up period (range, 7 months to 2 1/2 years). A diabetes like state was induced in five of the six dogs, and no nephrotoxicity was seen. Diabetes was not induced in one dog owing to caudal migration of an undersized balloon during the infusion; this also resulted in reversible renal damage. CONCLUSION: This simple technique is effective for inducing diabetes in dogs, and morbidity and mortality rates are lower than those reported in the literature with other described techniques. PMID- 11394540 TI - Correction of helical CT attenuation values with wide beam collimation: in vitro test with urinary calculi. AB - RATIONALE AND OBJECTIVES: Urinary calculi are now commonly detected with helical computed tomography (CT), and it has been proposed that stone composition can be determined from CT attenuation values. However, typical scans are made with a beam collimation of 5 mm or more, resulting in volume averaging and reduction in accuracy of attenuation measurement. The authors tested a model for correction of errors in attenuation values, even at section widths larger than the width of the object. MATERIALS AND METHODS: Human urinary stones were scanned with helical CT at different beam collimation widths. A computer model was used to predict the effect of beam width and stone size on accuracy of measured attenuation. RESULTS: At 3-mm collimation, the model corrected the attenuation readings with an underestimation of 12% +/- 1 (compared with values at 1-mm collimation; 127 stones; diameters of 1.7-11.3 mm). With attenuation measured at 10-mm collimation, the model underestimated the true value by 34% +/- 3 (103 stones), with a significant negative correlation with stone diameter on magnitude of error (diameters of 3.0-11.3 mm). Correlation of data from patient scans with subsequent in vitro scanning of the same stones confirmed the validity of the model, but corrected in vivo scans consistently yielded lower values for the stones than in vitro. CONCLUSION: Volume averaging effects on attenuation in helical CT are predictable in vitro for urinary calculi--and presumably for other roughly spherical structures--as long as section width does not excessively exceed the diameter of the structure. PMID- 11394541 TI - Evaluation of a spiral nitinol temporary inferior vena caval filter. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate the (a) ability of a prototype temporary inferior vena caval (IVC) filter to trap and retain emboli in an ex vivo flow circuit, (b) feasibility of filter placement and removal via a superficial vein in sheep, and (c) intermediate-term effects of the filter on the insertion vein and at the filter site. MATERIALS AND METHODS: In an iliocaval circuit, embolus capture with the prototype filter was compared to that with a Greenfield filter. In addition, prototype filters were placed into the infrarenal IVC in six sheep. Placement via a superficial venous route was initially attempted. Inferior vena cavography was performed weekly, and filters were removed after 2, 3, or 4 weeks (n = 2 each). Two weeks after the filters were removed, vena cavograms were obtained, the animals were sacrificed, and the IVC was evaluated at pathologic examination. RESULTS: The prototype filter captured all emboli, and the Greenfield filter captured 70%-100% of emboli. Successful placement via a superficial venous route was accomplished in only two sheep owing to small vein caliber; four filters were placed via a deep vein. Adverse events included perifilter thrombus, insertion site infection, and caudal migration. Two sheep died before filter removal owing to sepsis and anesthetic complications. The filters in the remaining four sheep were easily and successfully removed. Five sheep had stenosis at the filter site, and fibrosis with acute and chronic inflammation was seen at microscopic examination. CONCLUSION: The prototype filter trapped emboli as well as the Greenfield filter. Insertion via a superficial route, however, is possible only if the access vein is of an adequate size. PMID- 11394542 TI - Impact of unilateral common iliac vein occlusion on trapping efficacy of the Greenfield filter: an in vitro study. AB - RATIONALE AND OBJECTIVES: The purpose of this study was to assess the effect of unilateral common iliac vein occlusion on the capturing efficacy of the Greenfield filter in vitro. MATERIALS AND METHODS: A stainless steel over-the wire Greenfield filter was placed in the Silastic inferior vena cava module of a pulsatile circuit. Three 30-mm blood clots in sets of five were injected through the module's right iliac limb with the circuit in four experimental conditions: vertical position, both iliac limbs patent (VP); vertical position, left iliac limb occluded (VOC); horizontal position, both iliac limbs patent (HP); and horizontal position, left iliac limb occluded (HOC). Each experiment was repeated 15 times, resulting in 75 clots per condition and a total of 300 clot introductions. RESULTS: Clot trapping efficacy was 36 of 75 (48%) for VP, 41 of 75 (55%) for VOC, 32 of 75 (43%) for HP, and 26 of 75 (35%) for HOC. Cross comparisons of the four conditions revealed a marginally significant difference (P = .0138 with a corrected test-wise alpha = .0125) only between horizontal and vertical positions with unilateral common iliac limb occlusion. CONCLUSION: Unilateral common iliac vein occlusion decreases the capturing efficacy of the Greenfield filter in the horizontal position in vitro. In patients with unilateral common iliac vein occlusion, use of inferior vena cava filters with higher capturing efficacy may be considered. PMID- 11394543 TI - Displacement of coils into the lung during embolotherapy: clinical importance and follow-up with helical CT. AB - RATIONALE AND OBJECTIVES: The authors performed this study to evaluate the clinical importance and computed tomographic (CT) appearance of coils displaced into the lung during embolotherapy. MATERIALS AND METHODS: The authors retrospectively studied clinical charts and serial chest images from 25 consecutive patients after coil embolization. Chest radiography was performed in all patients, whereas helical chest CT was performed only in patients in whom dislocated coils were visible on chest radiographs. Coils were applied for the treatment of peripheral arteriovenous (AV) malformations and fistulas (n = 9), renal AV malformations or fistulas (n = 8), and primary or secondary tumors (n = 8). Clinical charts were analyzed for short- and long-term symptoms; chest radiographs and CT scans were reviewed for signs indicative of pulmonary infarction. RESULTS: None of the patients had clinical symptoms suggestive of pulmonary infarction. In two of the 25 patients (8%), displaced coils were seen in the pulmonary vasculature at chest radiography; these patients had been treated for renal AV fistula and peripheral AV fistula, respectively. One patient had two coils in the left hemithorax (upper and lower lobe), and the other patient had two coils in the right hemithorax (middle lobe). Neither of the patients had abnormalities suggestive of pulmonary infarction at helical CT. CONCLUSION: Chest radiography can help confirm the presence of coils displaced to the pulmonary vasculature during embolotherapy. Helical CT can also help rule out the presence of coil-associated pulmonary infarction. PMID- 11394544 TI - Academic radiology and doctor discontent: the good news and the bad news. PMID- 11394545 TI - Academic radiology: the real good news. PMID- 11394546 TI - Evaluating medical students on radiology clerkships in a filmless environment: use of an electronic test prepared from PACS and digital teaching collection images. AB - RATIONALE AND OBJECTIVES: The competency of medical students in radiology clerkships is traditionally evaluated with film images, projected slides of photographed films, or printed reproductions of films. As radiology departments switch to filmless imaging, it seemed appropriate to determine the feasibility of an electronic evaluation prepared directly from digital images. MATERIALS AND METHODS: The image-based portion of a multiple-choice final examination was prepared as a PowerPoint presentation that included images downloaded from the departmental picture archiving and communication system (PACS) or digital teaching collections. The images were downloaded as bitmap files, imported to Adobe Photoshop for image editing, converted to tagged image file format, and finally imported to PowerPoint, where they were combined with text to create 50 questions. A liquid crystal diode projector displayed the questions, with a timer set to advance them automatically. RESULTS: The examination was easy and inexpensive to prepare (no photography costs). In an initial survey of 25 students, 17 (71%) of 24 students rated the resolution of images as excellent and five (21%) as good. No student gave an image a poor rating. Students preferred that images cover at least 40%-50% of the slides, and most approved of a blue background. An original allowance of 30 seconds per slide was reported to be too fast; the interval was increased to 45 seconds. CONCLUSION: An electronic final examination for medical students, prepared with images downloaded from PACS or digital teaching collections, is feasible, easy to prepare, and cost-effective, and it provides an excellent display of test images. PMID- 11394547 TI - A detailed audit of reimbursement for abdominal CT in an academic practice. AB - RATIONALE AND OBJECTIVES: Declining fee schedules, decreasing operating margins, and increasingly stringent compliance regulations create a need for intense scrutiny and optimization of a radiology organization's billing and collection procedures. The authors' goal was to analyze the effectiveness of departmental professional billing procedures, identify controllable factors, and intervene when they could be improved. MATERIALS AND METHODS: A detailed audit of professional claims and payments was performed for all patients who underwent abdominal-pelvic computed tomography (CT) during July 1999 (n = 717). The adequacy of indication for the CT examination as given by the referring physician and modified by the radiology staff, the time required for claim generation, and the status of reimbursement within 120 days were assessed by an interdisciplinary team. After an intervention was performed to improve adequacy of the available clinical indication, the audit was repeated in December 1999 (n = 710). RESULTS: Despite a significant (P < .05) improvement in wording of clinical indications for billing purposes between July (68%) and December (85%), there was no significant change in reimbursement against gross charges. The vast majority of claims (97% in July, 99% in December) were generated in less than 30 days. At 120 days after the date of service, payments had been received that amounted to only 66% and 54% of discounted professional fees for July and December, respectively. For examinations performed in December, payment was delayed beyond contracted time periods in 138 cases (19%). CONCLUSION: Optimum billing and collection for imaging studies is an increasingly complex task. Even when substantial efforts are devoted to eliciting the proper indication for the study, reimbursement remains low primarily because of payer delays. PMID- 11394548 TI - Measuring the academic radiologist's clinical productivity: survey results for subspecialty sections. AB - RATIONALE AND OBJECTIVES: The purpose of this project was to understand better the academic radiologist's clinical workload in order to determine faculty staffing requirements more accurately. MATERIALS AND METHODS: Surveys performed by the Society of Chairmen of Academic Radiology Departments (SCARD) collected data for radiologists in 20 departments in 1996 and 1998; the data included work relative value units (RVUs) per full-time equivalent (FTE). Radiologists in each subspecialty were compared with their counterparts in other departments. The data were collected for each radiologist. Summary statistics showing averages, medians, and quartiles were used to describe workload (in RVUs per FTE) for each department and each subspecialty. RESULTS: Overall, the average clinical workload was 4,458 RVU/FTE, with 0.62 RVU per procedure. In those sections for which the faculty performed similar types of procedures across departments, the results were useful. The workload data, however, proved inadequate to compare across subspecialty sections. Between 1996 and 1998, the workload increased from 3,790 to 4,458 RVU/FTE. CONCLUSION: The SCARD survey provided very useful clinical workload data, measured in work RVUs per FTE for specific subspecialty sections. At practically all surveyed institutions, increasing clinical workload is competing with academic activities. PMID- 11394549 TI - Measuring the academic radiologist's clinical productivity: applying RVU adjustment factors. AB - RATIONALE AND OBJECTIVES: To improve understanding of academic radiologists' clinical workloads, the Society of Chairmen of Academic Radiology Departments (SCARD) performed surveys to collect workload data for radiologists in 20 departments; workload was measured in relative value units (RVUs) per full-time equivalent (FTE). Although they were useful for comparisons within some subspecialties, the workload data proved inadequate for comparisons across sections, and adjustment factors were needed for each Current Procedure Terminology (CPT) code. MATERIALS AND METHODS: All CPT codes for examinations were divided into groups with similar radiologist work effort. Focusing on radiologists who worked almost exclusively in each group, the authors created adjustment factors by using data from the individual radiologists at each institution. RESULTS: The adjustment factors are 0.50 for angiography, 0.58 for computed tomography and magnetic resonance imaging, and 1.0 for nuclear medicine, plain radiography, and special procedures (no adjustment needed for these groups). These factors are multiplied by the work RVUs for each examination to create the adjusted workload RVUs. CONCLUSION: The SCARD survey provided very useful clinical workload data, with workload measured in work RVUs per FTE for specific subspecialty sections. The new adjusted workload RVUs allow comparison of radiologists' workload across subspecialties. PMID- 11394550 TI - Organizing the National Institute of Biomedical Imaging and Bioengineering. Part 1. PMID- 11394551 TI - Cognitive appraisal biases: an approach to understanding the relation between socioeconomic status and cardiovascular reactivity in children. AB - We tested the hypothesis that lower socioeconomic status (SES) children display heightened cardiovascular reactivity during stressful situations because they are more likely to appraise a wide variety of social situations, including ambiguous ones, as threatening. A sample of 201 children and adolescents, half White and half African American, were assessed initially. Ninety of these children were retested an average of 3 years later. At both time points, children were interviewed about appraisals of hostile intent and feelings of anger in response to scenarios with negative or ambiguous outcomes. Cardiovascular reactivity to 3 laboratory stress tasks was measured. Initially, lower SES was associated with greater hostile intent appraisal and anger during ambiguous scenarios across all participants. In addition, responses to ambiguous scenalios partially mediated the relation between SES and vascular reactivity. Longitudinally, low SES African American participants showed higher mean intensity of hostile intent appraisals during ambiguous scenarios, and these appraisals mediated the SES-reactivity relationship. These findings suggest that the way in which children appraise ambiguous social situations plays an important role in the relation between SES and cardiovascular reactivity. PMID- 11394552 TI - A prospective study of the impact of stress on quality of life: an investigation of low-income individuals with hypertension. AB - The role that major and minor life events play in the quality of life in low income hypertensives was examined. Participants were randomly recruited from 2 primary care clinics at a public medical center. The study utilized a prospective design. Participants were determined to have hypertension and were being treated with antihypertensive medication prior to and throughout the duration of the study. Participants were administered the Life Experiences Survey and the Weekly Stress Inventory repeatedly during Year 1 to assess major and minor stress, respectively. Participants were repeatedly administered the RAND 36-Item Health Survey during Year 2 to assess quality of life. Usable data were obtained from 183 patients. Analyses revealed that major and minor stress were significant predictors of all measured domains of quality of life, even after age and number of chronic illnesses were statistically controlled. Minor stress contributed uniquely to the prediction of each dimension of quality of life even when age, number of chronic illnesses, and major life events were accounted for. Findings suggest that stress has a significant, persistent impact on the quality of life of low-income patients with established hypertension. These findings extend prior research that has examined the impact of medications on quality of life and suggest that stress needs to be accounted for as well. PMID- 11394553 TI - Public and private self-consciousness and smoking behavior in head and neck cancer patients. AB - Patients who continue to use tobacco following treatment for head and neck cancers are at a greater risk for cancer recurrence and earlier mortality. This study examined the unique effects of public and private self-consciousness and negative affect on smoking behavior in a sample of 40 patients with cancers of the head and neck. Measures of public and private self-consciousness and negative affect were administered and assessments of past and current smoking behavior were obtained. Only public self-consciousness was a significant predictor of continued smoking following oncologic treatment. Specifically, individuals with low levels of public self-consciousness were nearly 13 times more likely to continue smoking compared to those with relatively higher levels of public self consciousness. This pattern is interpreted in the context of previous theorizing that suggests individuals high in public self-consciousness are more likely to discontinue habitual behavior that is perceived as socially undesirable or incorrect. PMID- 11394554 TI - Injectable medication for the treatment of multiple sclerosis: the influence of self-efficacy expectations and injection anxiety on adherence and ability to self inject. AB - The management of many chronic illnesses involves medications that must be injected on a frequent basis. With fewer support resources available, patients are increasingly being obliged to manage injectable medications themselves. Interferon beta-1a (IFNbeta-1a), recommended for the treatment of multiple sclerosis (MS), must be injected intramuscularly on a weekly basis. Patients are generally advised and taught to self-inject, if possible. This longitudinal study examined cognitive and affective contributions to the ability to self-inject and adherence to IFNbeta-1a over 6 months following initiation of medication. Participants were 101 patients with a relapsing form of MS. Injection self efficacy expectations, injection anxiety, adherence expectations, method of injection administration, and 6-month adherence to IFNbeta-1a were fitted to a path analytic model. Pretreatment injection self-efficacy expectations were significantly related to 6-month adherence. This relation was mediated by the patient's ability to self-inject. Patients 'experienced level of injection anxiety was related to adherence but not to method of injection. PMID- 11394555 TI - Distress and disease status among patients with rheumatoid arthritis: roles of coping styles and perceived responses from support providers. AB - Previous research has shown that social support can have a beneficial impact on coping processes and psychological adjustment in patients with rheumatoid arthritis (RA). The association of individual coping styles and perceived responses from others to one's pain episodes with patients 'distress and disease status over time was investigated. The sample consisted of 42 middle-aged patients with RA who were predominantly White (98%), female (64%), and married (88%). Participants completed surveys and their rheumatologist completed clinical assessments of patient disease status at 2 time points over a 9-month period. Although punishing responses from others (e.g., getting irritated or angry when the patient is in pain) were perceived as relatively infrequent, they were associated with a patient coping style of focusing on and venting of negative emotion as well as elevated negative affect (NA). Findings also indicated that those who perceived punishing responses from close others and coped by venting negative emotions reported increased NA over time and were rated by their rheumatologist as having more severe RA disease status over time. Implications for psychosocial intervention and directions for future research are discussed. PMID- 11394556 TI - An evaluation of the effects of Tai Chi exercise on physical function among older persons: a randomized contolled trial. AB - This study was designed to determine whether a 6-month Tai Chi exercise program can improve self-reported physical functioning limitations among healthy, physically inactive older individuals. Ninety-four community residents ages 65 to 96 (Mage = 72.8 years, SD = 5.1) volunteered to participate in the study. Participants were randomly assigned to either a 6-month experimental (Tai Chi) group (n = 49), which exercised twice per week for 60 min, or a wait-list control group (n = 45). A 6-item self-report physical functioning scale, assessing the extent of behavioral dysfunction caused by health problems, was used to evaluate change in physical functioning limitations as a result of Tai Chi intervention. Results indicated that compared to the control group, participants in the Tai Chi group experienced significant improvements in all aspects of physical functioning over the course of the 6-month intervention. Overall, the experimental group had 65% improvement across all 6 functional status measures ranging from daily activities such as walking and lifting to moderate-vigorous activities such as running. It was concluded that the 6-month Tai Chi exercise program was effective for improving functional status in healthy, physically inactive older adults. A self-paced and self-controlled activity such as Tai Chi has thepotential to be an effective, low-cost means of improving functional status in older persons. PMID- 11394557 TI - Progress in behavioral research on physical activity. PMID- 11394558 TI - Maintenance of physical activity following an individualized motivationally tailored intervention. AB - This study examined predictors of exercise maintenance following completion of a physical activity intervention. Sedentary adults recruited through newspaper advertisements were randomly assigned to receive either (a) a motivation-matched intervention with feedback reports that were individually tailored (IT) to psychological variables from social cognitive theory and the Transtheoretical Model via computer expert system, or (b) a standard, print-based intervention (ST). The intervention phase of the study included mailed assessments and intervention materials at baseline, 1, 3, and 6 months. An assessment-only follow up was conducted 6 months after the end of the intervention (Month 12). Participants were assessed for current physical activity participation, motivational readiness for physical activity, a number of psychological constructs posited to influence participation in physical activity (e.g., self efficacy), and current affect. Significantly more participants in the IT condition met or exceeded exercise participation goals at the end of the intervention period and maintained this level of physical activity through the Month 12 follow-up compared to ST participants. Prospective analyses revealed significant differences in several psychological constructs both at program entry (baseline) and the end of the intervention period between individuals who maintained their physical activity participation through Month 12 and those who did not. Results suggest that the maintenance of physical activity following the end of an active intervention program may be influenced by attitudes and behaviors acquired along with increased participation in physical activity, as well as by preexisting characteristics that individuals bring into treatment. PMID- 11394559 TI - A computerized social cognitive intervention for nutrition behavior: direct and mediated effects on fat, fiber, fruits, and vegetables, self-efficacy, and outcome expectations among food shoppers. AB - This study examined the direct and mediated impact of a self-administered, computer-based intervention on nutrition behavior self-efficacy, and outcome expectations among supermarket food shoppers. The intervention, housed in kiosks in supermarkets and based on social cognitive theory, used tailored information and self-regulation strategies delivered in 15 brief weekly segments. The study sample (N = 277), stratified and randomly assigned to treatment or control, was 96% female, was 92% White, had a median annual income of about $35,000, and had a mean education of 14. 78 +/- 2.11 years. About 12% of the sample reported incomes of $20,000 or less, and about 20% reported 12 years or fewer of education. Analysis of covariance immediately after intervention and at a 4- to 6-month follow-up found that treatment led to improved levels of fat, fiber, and fruits and vegetables. Treatment also led to higher levels of nutrition-related self efficacy, physical outcome expectations, and social outcome expectations. Logistic regression analysis determined that the treatment group was more likely than the control group to attain goals for fat, fiber, and fruits and vegetables at posttest and to attain goals for fat at follow-up. Latent variable structural equation analysis revealed self-efficacy and physical outcome expectations mediated treatment effects on nutrition. In addition, physical outcome expectations mediated the effect of self-efficacy on nutrition outcomes. Implications for future computer-based health promotion interventions are discussed. PMID- 11394560 TI - Assessing patients with possible heart disease using scores. AB - Multivariable analysis of clinical and exercise test data has the potential to become a useful tool for assisting in the diagnosis of coronary artery disease, assessing prognosis, and reducing the cost of evaluating patients with suspected coronary disease. Since general practitioners are functioning as gatekeepers and decide which patients must be referred to the cardiologist, they need to use the basic tools they have available (i.e. history, physical examination and the exercise test), in an optimal fashion. Scores derived from multivariable statistical techniques considering clinical and exercise data have demonstrated superior discriminating power compared with simple classification of the ST response. In addition, by stratifying patients as to probability of disease and prognosis, they provide a management strategy. While computers, as part of information management systems, can run complicated equations and derive these scores, physicians are reluctant to trust them. Thus, these scores have been represented as nomograms or simple additive tables so physicians are comfortable with their application. Their results have also been compared with physician judgment and found to estimate the presence of coronary disease and prognosis as well as expert cardiologists and often better than nonspecialists. However, the discriminating power of specific variables from the medical history and exercise test remains unclear because of inadequate study design and differences in study populations. Should expired gases be substituted for estimated metabolic equivalents (METs)? Should ST/heart rate (HR) index be used instead of putting these measurements separately into the models? Should right-sided chest leads and HR in recovery be considered? There is a need for further evaluation of these routinely obtained variables to improve the accuracy of prediction algorithms especially in women. The portability and reliability of these equations must be demonstrated since access to specialised care must be safe-guarded. Hopefully, sequential assessment of the clinical and exercise test data and application of the newer generation of multivariable equations can empower the clinician to assure the cardiac patient access to appropriate and cost-effective cardiological care. PMID- 11394561 TI - Neural influences on sprint running: training adaptations and acute responses. AB - Performance in sprint exercise is determined by the ability to accelerate, the magnitude of maximal velocity and the ability to maintain velocity against the onset of fatigue. These factors are strongly influenced by metabolic and anthropometric components. Improved temporal sequencing of muscle activation and/or improved fast twitch fibre recruitment may contribute to superior sprint performance. Speed of impulse transmission along the motor axon may also have implications on sprint performance. Nerve conduction velocity (NCV) has been shown to increase in response to a period of sprint training. However, it is difficult to determine if increased NCV is likely to contribute to improved sprint performance. An increase in motoneuron excitability, as measured by the Hoffman reflex (H-reflex), has been reported to produce a more powerful muscular contraction, hence maximising motoneuron excitability would be expected to benefit sprint performance. Motoneuron excitability can be raised acutely by an appropriate stimulus with obvious implications for sprint performance. However, at rest H-reflex has been reported to be lower in athletes trained for explosive events compared with endurance-trained athletes. This may be caused by the relatively high, fast twitch fibre percentage and the consequent high activation thresholds of such motor units in power-trained populations. In contrast, stretch reflexes appear to be enhanced in sprint athletes possibly because of increased muscle spindle sensitivity as a result of sprint training. With muscle in a contracted state, however, there is evidence to suggest greater reflex potentiation among both sprint and resistance-trained populations compared with controls. Again this may be indicative of the predominant types of motor units in these populations, but may also mean an enhanced reflex contribution to force production during running in sprint-trained athletes. Fatigue of neural origin both during and following sprint exercise has implications with respect to optimising training frequency and volume. Research suggests athletes are unable to maintain maximal firing frequencies for the full duration of, for example, a 100m sprint. Fatigue after a single training session may also have a neural manifestation with some athletes unable to voluntarily fully activate muscle or experiencing stretch reflex inhibition after heavy training. This may occur in conjunction with muscle damage. Research investigating the neural influences on sprint performance is limited. Further longitudinal research is necessary to improve our understanding of neural factors that contribute to training-induced improvements in sprint performance. PMID- 11394564 TI - High speed immuno-affinity chromatography on supports with gigapores and porous glass. AB - Immuno-affinity chromatography exploiting the Ca2+ dependent interaction of the anti-Flag antibody and Flag-tagged proteins has been investigated. The antibody has been immobilized on porous glass beads (Prosep) containing gigapores and on a monolith, the polymethacrylate based Convective Interactive Media (CIM) column at a ligand density of 2 mg/g and 10 mg/ml respectively. The performance of the columns was assessed by applying clarified yeast culture supernatant containing overexpressed Flag-human serum albumin. Dynamic binding capacity and purity was checked at various flow rates ranging from 100 cm/h to 800 cm/h. 95% purity could be obtained. Anti Flag-CIM columns showed a higher unspecific adsorption, requiring a longer wash cycle to obtain the same purity compared to the Prosep column. Anti Flag-CIM columns showed a flow independent performance, which is explained by its monolithic structure. A decreasing dynamic binding capacity with flow was observed with anti-Flag-Prosep columns. Both columns are suited to purify milligrams of protein out of a yeast culture supernatant within a few minutes. We considered them as promising candidates for high throughput screening, where fast purification is a necessity. PMID- 11394562 TI - Exercise in the prevention of falls in older people: a systematic literature review examining the rationale and the evidence. AB - Falls are a major source of death and injury in elderly people. For example, they cause 90% of hip fractures and the current cost of hip fractures in the US is estimated to be about 10 billion dollars. Age-related changes in the physiological systems (somatosensory, vestibular and visual) which contribute to the maintenance of balance are well documented in older adults. These changes coupled with age-related changes in muscle and bone are likely to contribute to an increased risk of falls in this population. The integrated rehabilitation based model of fall risk factors reveals multiple sites for interventions that may reverse fall risk factors. Regular exercise may be one way of preventing falls and fall-related fractures. The evidence for this contention comes from a variety of sources. On the basis of 9 randomised controlled studies conducted since 1996, exercise appears to be a useful tool in fall prevention in older adults, significantly reducing the incidence of falls compared with control groups. However, current limitations such as inconsistencies in the measurement of key dependent and independent variables do not, at present, permit a meta analysis of intervention trials. Further investigation, using trials designed with the current limitations in mind, is necessary to establish the optimum exercise programme to maximise fall prevention in older adults. PMID- 11394563 TI - Physical activity assessment in children and adolescents. AB - Chronic disease risk factors, including a sedentary lifestyle, may be present even in young children, suggesting that early prevention programmes may be critical to reducing the rates of chronic disease. Accurate assessment of physical activity in children is necessary to identify current levels of activity and to assess the effectiveness of intervention programmes designed to increase physical activity. This article summarises the strengths and limitations of the methods used to evaluate physical activity in children and adolescents. MEDLINE searches and journal article citations were used to locate 59 articles that validated physical activity measurement methods in children and adolescents. Only those methods that were validated against a more stringent measure were included in the review. Based on the definition of physical activity as any bodily movement resulting in energy expenditure (EE), direct observation of the individual's movement should be used as the gold standard for physical activity research. The doubly labelled water technique and indirect calorimetry can also be considered criterion measures for physical activity research, because they measure EE, a physiologic consequence closely associated with physical activity. Devices such as heart rate monitors, pedometers and accelerometers have become increasingly popular as measurement tools for physical activity. These devices reduce the subjectivity inherent in survey methods and can be used with large groups of individuals. Heart rate monitoring is sufficiently valid to use in creating broad physical activity categories (e.g. highly active, somewhat active, sedentary) but lacks the specificity needed to estimate physical activity in individuals. Laboratory and field validations of pedometers and accelerometers yield relatively high correlations using oxygen consumption (r = 0.62 to 0.93) or direct observation (r = 0.80 to 0.97) as criterion measures, although, they may not be able to capture all physical activity. Physical activity has traditionally been measured with surveys and recall instruments. These techniques must be used cautiously in a paediatric population that has difficulty recalling such information. Still, some studies have reported 73.4% to 86.3% agreement between these instruments and direct observation. Future investigations of physical activity instruments should validate the novel instrument against a higher standard. Additional studies are needed to investigate the possibility of improving the accuracy of measurement by combining 2 or more techniques. The accurate measurement of physical activity is critical for determining current levels of physical activity, monitoring compliance with physical activity guidelines, understanding the dose-response relationship between physical activity and health and determining the effectiveness of intervention programmes designed to improve physical activity. PMID- 11394565 TI - Extraction of peptide tagged cutinase in detergent-based aqueous two-phase systems. AB - Detergent-based aqueous two-phase systems have the advantage to require only one auxiliary chemical to induce phase separation above the cloud point. In a systematic study the efficiency of tryptophan-rich peptide tags was investigated to enhance the partitioning of an enzyme to the detergent-rich phase using cutinase as an example. Up to 90% enzyme activity could be extracted in a single step from whole broth of recombinant Saccharomyces cerevisiae expressing cutinase variants carrying a (WP)4 tag. In contrast, the extraction yield of wild type cutinase was 2-3% only. The detergent concentration and the temperature are the main parameters to optimize the extraction yield. Considering availability, extraction yields, and price the detergent Agrimul NRE 1205 served best for enzyme recovery. PMID- 11394566 TI - Optimization of bovine serum albumin partition coefficient in aqueous two-phase systems. AB - Partitioning of proteins in aqueous two-phase systems has been shown to provide a powerful method for separating and purifying mixtures of biomolecules by extraction. These systems are composed of aqueous solutions of either two water soluble polymers, usually polyethylene glycol (PEG) and dextran (Dx), or a polymer and a salt, usually PEG and phosphate or sulfate. There are many factors which influence the partition coefficient K, the ratio of biomolecule concentration in the top phase to that in the bottom phase, in aqueous two-phase systems. The value of the partition coefficient relies on the physico-chemical properties of the target biomolecule and other molecules and their interactions with those of the chosen system. In this work, the partition behavior of pure bovine serum albumin in aqueous two-phase systems was investigated in order to see the effects of changes in phase properties on the partition coefficient K. The concentration of NaCl and pH were considered to be the factors having influence on K. Optimal conditions of these factors were obtained using the Box Wilson experimental design. The optimum value of K was found as 0.0126 when NaCl concentration and pH were 0.14 M and 9.8, respectively, for a phase system composed of 8% (w/w) polyethylene glycol 3,350-9 (% w/w) dextran 37,500-0.05 M phosphate at 20 degrees C. PMID- 11394567 TI - Modeling of direct recovery of lactic acid from whole broths by ion exchange adsorption. AB - Lactic acid fermentation process with L. casei CRL 686 was performed. The static adsorption isotherm over a strong anionic exchange resin, Amberlite IRA-400 was measured, and the static binding capacity parameters were quantified. Early recovery of lactic acid from this lactate producer from unclarified culture broth was performed in a liquid solid fluidized bed, with the resin as the solid adsorbent, and the dynamic adsorption capacity was calculated. Good agreement was found between static and dynamic binding capacity values. The fluidized bed height was twice the settled bed height and the overall process was controlled by the liquid solid mass transfer. This operation was also simulated by continuously well stirred tanks arranged in series and superficial solid deactivation as in a gas solid catalytic reactor. The deactivation process takes into account liquid channeling and agglomerations of solid induced by the viscosity of the broth and also by the cells during the adsorption. These patterns were also verified by experimental observations, and are in agreement with the results found in the literature. The breakthrough data together with others from previous works were satisfactorily fitted until the 90% dimensionless concentration was reached for both culture broths. The model could be used in future studies on predictions about the liquid solid fluidized bed behavior and other different operating conditions. PMID- 11394568 TI - Recovery of a monoclonal antibody from hybridoma culture supernatant by affinity precipitation with Eudragit S-100. AB - An IgG1 monoclonal antibody (MAB) was isolated from hybridoma culture supernatant by affinity precipitation with an Eudragit S-100-based heterobifunctional ligand. Affinity binding was performed in a homogeneous aqueous phase at pH 7.5 followed by precipitation of the bound affinity complex by lowering the pH to 4.8. After two washing steps, elution of specifically bound MAB was achieved by incubating the precipitate with 0.1 M glycine.HCl pH 2.5. The influence of elution volume and time on the recovery of active MAB and the overall purification factor were studied. The best conditions enabled the recovery of 50.2% of active MAB with a purification factor of 6.2. A further dialysis against 50 mM Tris.HCl pH 8.0 increased the activity yield and the purification factor to 68.4% and 8.3, respectively. This result showed that part of the antibody activity loss during affinity precipitation was due to a reversible inactivation process, being easily recovered after a refining dialysis step. PMID- 11394569 TI - Eicosapentaenoic and arachidonic acids purification from the red microalga Porphyridium cruentum. AB - The polyunsaturated fatty acids (PUFA) eicosapentaenoic and arachidonic acids (EPA and AA), which have several pharmaceutical properties, have been purified from the red microalga Porphyridium cruentum. The process consists of only four main steps: (i) simultaneous extraction and saponification of the microalgal biomass; (ii) urea inclusion method (iii) PUFA esterification (iv) argentated silica gel column chromatography of the urea concentrate. Total AA and EPA recoveries reached 39.5% and 50.8% respectively for a purity approximately 97% for both fatty acids. Therefore, recovery of highly pure PUFA could be improved in organisms that are rich in two or more fatty acids of interest. The results of several procedures for AA and EPA recovery from several authors by using this microalga were compared. PMID- 11394570 TI - A simple and effective separation and purification procedure for DNA fragments using dodecyltrimethylammonium bromide. AB - In this work we describe a simple two step separation procedure for the separation and purification of short DNA fragments. The first step involves precipitating the DNA using the cationic surfactant dodecyltrimethylammonium bromide. Dodecyltrimethylammonium bromide, unlike cetyltrimethylammonium bromide will not precipitate DNA before complexation is complete thus providing a high purity DNA. The second step involves dissolution of the DNA dodecyltrimethylammonium complex in 75% ethanol, followed by precipitation of the Sodium-DNA salt, by titrating in a salt solution. This method is particularly suited to purification of short fragments as it does not require high salt concentrations in the ethanol precipitation step, which can be damaging for short DNA. The ability of dodecyltrimethylammonium bromide to remove ethidium bromide from intercalation sites on the DNA is also discussed PMID- 11394571 TI - Affinity chromatography of neoglycoproteins. AB - Glycoproteins, as a class of biomolecules, exhibit much more heterogeneous structures than non-glycosylated proteins. They present a challenging area of research. Model glycoproteins with well-defined protein and carbohydrate structures are helpful in the search for high-resolution methods for the separation of glycoproteins. Neoglycoproteins, maltose-modified chymotrypsin and lactose-modified chymotrypsin, were synthesised by modifying chymotrypsin with maltose and lactose, respectively, using the reductive amination method. Boronate chromatography was applied to isolate the neoglycoproteins from non-glycosylated substances. The use of Tris-HCl as a shielding reagent during the boronate chromatography proved to be efficient in eliminating unwanted interactions between the boronate ligand and the peptide backbone of chymotrypsin. The retention time of neoglycoproteins on the boronate column was increased with increasing the degree of modification. PMID- 11394572 TI - Predominance of antibodies to hepatitis C virus envelope proteins in various disease statuses of hepatitis C. AB - The antibody profile to various proteins of hepatitis C virus (HCV) was studied in 113 patients positive for HCV RNA in various disease statuses of hepatitis C (HC). A single peptide (E2/NS1, aa 413-436 of HCV polyprotein) chosen from a conserved region at the C-terminus of the hypervariable region (HVR) HVR1 of HCV was found to be sufficient for reliable diagnosis of the infection, even in the acute phase. Six hundred and one suspected HC cases and 200 voluntary blood donors were tested by this peptide. The sensitivity of detection of HCV antibodies by this peptide did not increase with addition of peptides from other HCV proteins. Our results clearly demonstrate that antibodies to HCV envelope proteins occur in a higher percentage of the infected population than those to other proteins. This emphasizes the necessity of using representative sequences from HCV envelope proteins in diagnostic immunoassays of this viral infection. PMID- 11394573 TI - Genetic and antigenic analysis of type A foot-and-mouth disease viruses isolated in India during 1987-1996. AB - Twenty-three foot-and-mouth disease virus (FMDV) type A field isolates, recovered from different outbreaks during 1987-1996 in India, were subjected to antigenic and genetic analysis. The isolates showed a close antigenic relationship to the current vaccine strain (IND 17/77) in micro-neutralization test conducted using a vaccine strain (IND 17/77) antiserum and a peptide (aa 136-151 of VP1 protein of the A22/Azerbaijan/65 strain) antiserum. However, the isolates revealed minor antigenic differences in their reactivity with three neutralizing monoclonal antibodies (MAbs) recognizing trypsin-sensitive conformation-independent epitopes of the vaccine virus strains. Phylogenetic relationship between the isolates was carried out employing a part of the 1D gene (168 nucleotides at the 3'-end). Additional seven type A Indian field isolates reported earlier were included in the analysis. The percent similarity among the Indian isolates varied from 82.7% to 99.4% at nucleotide level, and from 83.9% to 100% at amino acid level. These observations clearly demonstrate genetic heterogeneity of the field isolates. The current vaccine strain IND 17/77 showed divergence of 9.7% at nucleotide level and 5.6% at amino acid level from the A22 Iraq 24/64 isolate. The field strains were divergent from the vaccine strain IND 17/77 by 5.6%-14.6% and 3.7% 13.7% at nucleotide and amino acid level, respectively. In the phylogenetic tree, the isolates were distributed into 21 genetic groups. The clustering pattern of the isolates in the phylogenetic tree revealed no specific distribution pattern of the foot-and-mouth disease (FMD) outbreaks in relation to their geographical locations, caused by unrestricted animal movement and endemic nature of the disease. PMID- 11394574 TI - Oral immunization of pigs against classical swine fever. Course of the disease and virus transmission after simultaneous vaccination and infection. AB - The efficacy of simultaneous vaccination of pigs against classical swine fever (CSF) and challenge was evaluated. In this study, domestic weanling pigs were vaccinated orally with a conventional live virus vaccine based on CSF virus (CSFV) C strain and were challenged simultaneously with CSFV of different virulence. All the animals vaccinated and challenged with a high dose of highly virulent Koslov strain died while three of five animals challenged with a low dose of highly virulent Alfort 187 strain survived, shed the virus in nasal secretions, developed antibodies, and four of them showed a transient viremia. All the animals vaccinated and challenged with the low virulent field isolate MV 140/Riems survived, showed a short viremia and developed antibodies. No CSFV or CSFV RNA could be detected in the animals surviving the infection. This study demonstrates that oral vaccination of wild boars in an infected area bears no risk for the development of a persistent CSF infection. PMID- 11394575 TI - Nucleocapsid protein gene sequence analysis reveals close genomic relationship between turkey coronavirus and avian infectious bronchitis virus. AB - Antibodies to infectious bronchitis virus (IBV) cross-react with turkey coronavirus (TCV) in immunofluorescence assay (IFA) indicating that IBV and TCV may share an amino acid sequence similarity. To determine its extent, the gene encoding the nucleocapsid (N) protein of TCV was amplified by reverse transcription-PCR (RT-PCR) from RNA purified from intestines of embryos of turkeys infected with various TCV isolates and from allantoic fluid of chicken embryos infected with IBV M41 strain, the obtained N genes were cloned, sequenced and compared with known sequences of N genes of five IBV strains. The primers for amplification were designed from the genome of IBV PCR products were obtained only from two of eight TCV isolates tested. It was found that the two TCV isolates were identical with five IBV strains by 90.1-94.1% at the N gene level. It was also observed that the N gene of eight TCV isolates originating from various regions of the USA could not be amplified by the primers designed from the N gene of bovine coronavirus (BCV). PMID- 11394576 TI - Influenza vaccination of human immunodeficiency virus 1-infected patients receiving antiretroviral therapy. AB - In 13 human immunodeficiency virus 1 (HIV-1) infected patients receiving a highly active antiretroviral therapy (HAART) annual influenza vaccination was conducted. It was hoped that HAART would prevent a post-vaccination increase in HIV-1 load and potential adverse effects. Only two patients had an increased viral load on day 14 post vaccination (p.v.). At 6 months p.v., the majority of the patients had a significantly increased CD4 cell count and a significantly decreased viral load. This indicates that HAART can protect patients from adverse consequences of influenza vaccination. The production of antibodies to the influenza A and B viruses in the HIV-infected patients was substantially lower than that in healthy persons. We propose that HIV-positive patients receiving HAART should be subjected to annual influenza vaccination. PMID- 11394577 TI - Molecular characterization of a novel defective DNA isolated from tobacco tissues infected with tobacco leaf curl virus. AB - Defective DNA of tobacco leaf curl virus (TLCV) was identified in TLCV-infected tobacco plants. The defective DNA was cloned and sequenced. The sequence showed it was about half the size of the TLCV DNA-A, and was derived from TLCV DNA-A by a large deletion. The defective DNA contained the intergenic region and part of the AC1 (Rep) gene of TLCV, and also novel open reading frames (ORFs). The immunotrapping tests showed the defective DNA was associated with geminate particles, suggesting it could be encapsidated in virus particles. It was transmitted, along with full-length DNA-A, to tobacco plants by grafting and whitefly (Bemisia tabaci). PMID- 11394578 TI - Glycyrrhizin increases survival of mice with herpes simplex encephalitis. AB - We examined the effect of glycyrrhizin (GR), a component of licorice root extract, on herpetic encephalitis that was inflicted on mice by inoculation of herpes simplex virus 1 (HSV-1) onto their cornea. Intraperitoneal (i.p.) administration of GR to mice suffering from herpetic encephalitis increased their survival rate in average about 2.5 times (from 37.5-29.0% to 81.8-83.3%; mean values from 2 experiments) while it reduced HSV-1 replication in the brain to 45.6% of the control. These results demonstrate a stimulative effect of GR on the mouse defense system(s) against HSV-1 infection. PMID- 11394579 TI - Nucleotide sequences of 5'-terminal parts of coat protein genes of various isolates of NTN strain of potato virus Y. AB - Sequences of the first 300 nucleotides of coat protein (CP) genes of 7 isolates of NTN strain of potato virus Y (PVY, PVY(NTN)) were determined and compared with analogous published sequences of various isolates and strains of PVY. The sequence identity among the sequenced isolates ranged from 96 to 100%. The differences were found at different positions. The nucleotide sequence of this part of CP gene seems to be very conservative among the isolates tested that means that PVY(NTN) is the evolutionary youngest among all PVY strains. PMID- 11394580 TI - Preparation of recombinant coat protein of Prunus necrotic ringspot virus. AB - The coat protein (CP) gene of Prunus necrotic ringspot virus (PNRSV) was cloned into pET 16b vector and expressed in Escherichia coli. CP-enriched fractions were prepared from whole cell lysate by differential centrifugation. The fraction sedimenting at 20,000 x g for 30 mins was used for preparation of a rabbit antiserum to CP. This antiserum had a titer of 1:2048 and reacted in a double antibody sandwich ELISA (DAS-ELISA). PMID- 11394581 TI - The presence or absence of ribonucleotide reductase activity has no affect on avirulent bovine herpesvirus 2. PMID- 11394582 TI - Autoantibodies to asialoglycoprotein receptor in chronic hepatitis C patients. AB - The aim of this study was to determine the incidence of autoantibodies to asialoglycoprotein receptor (ASGPR, anti-ASGPR) in chronic hepatitis C patients and to characterize the anti-ASGPR-positive and anti-ASGPR-negative patients in more detail. A total of 79 chronic hepatitis C patients were screened for the presence anti-ASGPR by ELISA. Anti-ASGPR were detected in 11 (13.9%) patients. No significant differences were found between the anti-ASGPR-positive and anti-ASGPR negative patients in age, alanine transaminase (ALT) activity, histological findings and response and tolerance to alpha-interferon (alpha-IFN) therapy. The male predominance in the anti-ASGPR positive group was statistically significant. It was surprising that other tested autoantibodies (antinuclear autoantibodies [ANA], smooth muscle autoantibodies [SMA], type 1 liver-kidney microsome autoantibodies [LKM-1], anti-thyroglobulin and thyroid microsome autoantibodies) and increased levels of immunoglobulins A, G and/or M were observed significantly more frequently in the anti-ASGPR-negative group. PMID- 11394583 TI - Weighted health status in the Medicare population: development of the Weighted Health Index for the Medicare Current Beneficiary Survey (WHIMCBS). AB - We present an approach to constructing an aggregate index of health at the population level with data from Medicare beneficiaries using the 1991 (N = 12,667), 1995 (N = 15,590), and 1997 (N=17,058) Medicare Current Beneficiary Survey (MCBS). Similar to other work with survey data, we develop a weighted health status index from which one can calculate a point in time health status score for any beneficiary. Scores range from 1.0, representing "excellent health and no activity limitation", to 0.0, representing deceased. Sequences of numerically weighted health states experienced over time can be summed to calculate years of healthy life for beneficiaries. We test both the stability of the scoring system when developed on independent samples, as well as the sensitivity of years of healthy life calculations to changes in scoring assumptions. Findings suggest that, in addition to mortality, morbidity appears to play a significant role in the years of healthy life accrued by Medicare beneficiaries since entry into the Medicare program. Further, the index scoring system is highly stable when derived on independent samples. Finally, calculations of years of healthy life are robust to changes in scoring assumptions. The weighted health index for Medicare current beneficiaries (WHIMCBS) is a stable overall index of health and may be a useful ongoing indicator of health within the Medicare population. PMID- 11394584 TI - ECRHS screening questionnaire scoring: a methodological suggestion for asthma assessment. European Community Health Survey. AB - Univocal definition and classification of Asthma have always been a matter of discussion, and that is reflected in the difficulty of constructing a measure of pathology severity. The European Community Health Survey is a multinational survey designed to compare the prevalence of asthma in subjects, aged 20 to 44 years, in several European areas. In each participating center a sample of 3000 adults filled a self-administered screening questionnaire composed by 9 dichotomous items. Aim of the present study is to investigate unidimensionality of the ECRHS screening questionnaire and to determine and validate a scoring of asthma-like symptoms seriousness. Dimensionality and scoring was determined through a Homogeneity Analysis by Alternating Least Square; while scoring validation was assessed by a cross validation technique. This study found the existence of a sole dimension underlying the screening questionnaire; furthermore a scoring of asthma-like symptoms seriousness was determined with the indication of a cut-off in order to distinguish between asthma symptomatic and non symptomatic subjects. PMID- 11394585 TI - Measurement Properties of the Symptom Impact Inventory. AB - The purpose of this study is to evaluate the measurement properties of the Symptom Impact Inventory using both psychometric and Rasch analyses. This inventory is designed for generally healthy midlife women. The sample included 340 midlife women aged 45-65 representing two studies. The first study involved Black and White employed sedentary women (n = 161) who volunteered for a walking intervention. The second study of migration and health included women who were recent immigrants from the former Soviet Union (n = 179). The women reported experiencing an average of 13.44 symptoms (S.D.=7.88) with a range of 1 to 32. Principal components analysis identified 5 components in this sample. Rasch measurement analysis found excellent model fit for the Symptom Impact Inventory with only 2 symptoms, Decreased appetite and Decreased sexual desire or interest, unstable in scale dimensionality analyses. Person and item parameters were reliable, and comparisons with groups known to differ on symptom reporting provided substantial validity. Although the two sample groups differed significantly on most demographic characteristics, a cross-cultural comparison found the scale structure remarkably robust. PMID- 11394586 TI - Prevention of secondary health conditions in adults with developmental disabilities: a review of the literature. AB - PURPOSE: To compile empirical findings regarding prevention strategies for secondary conditions experienced by adults with developmental disabilities. METHOD: The PsycLit and PubMed databases were searched for articles addressing the 20 most pervasive secondary conditions and the prevention of secondary conditions in general. RESULTS: Of more than 2000 articles examined, 25 met criteria for inclusion. None could be categorized as a primary approach to prevention, 19 involved secondary and six involved tertiary approaches. The majority included between one and ten participants. Nine studies involved the administration of treatment, three providing certain experiences, two chart review, and 11 a training approach. Injuries due to self-abuse, communication problems, dental hygiene problems, and problems with memory, persistence and mobility were addressed. CONCLUSION: Little is known regarding the prevention of secondary conditions within this population. The authors stress the necessity to focus research efforts on greater understanding of the linkage between disability, rehabilitation and public health models. PMID- 11394587 TI - The impact of traumatic brain injury on family members living with patients: a preliminary study in Japan and the UK. AB - PURPOSE: To ascertain the views of families living with TBI patients about the nature of the problems experienced as a result of TBI, and to compare the views of Japanese family members (J-FM) and British family members (B-FM) in order to find out whether there were cultural differences in family response to TBI. METHODS: Family members involved in providing care were identified by the patients. Face to face interviews were conducted with all 18 carers in B-FM and four carers in J-FM. The remaining eight carers in J-FM participated in the postal questionnaire. Questionnaires were developed to explore the nature of problems and the involvement of family such as social embarrassment. RESULTS: Problems arising in families were almost the same reported from both groups. However families in B-FM were likely to know more about how to cope with these problems. Family members in J-FM reported more statistically significant increases in social embarrassment than those in B-FM. CONCLUSION: The preliminary results showed that family members living with TBI patients in both groups had experienced problems. Appropriate rehabilitation services should be developed to help families as well as TBI patients in Japan. PMID- 11394588 TI - Knee muscle isometric strength, voluntary activation and antagonist co contraction in the first six months after stroke. AB - PURPOSE: Muscle weakness may contribute to functional problems after stroke, but is rarely addressed during rehabilitation. Functional problems are commonly thought to be caused by abnormal movement patterns or possibly disuse atrophy. We investigated voluntary isometric strength, activation and the extent of co contraction in the knee muscles during the first six months during stroke. METHODS: Twelve stroke patients (58 +/- 3 years, mean+/- SEM, 7 female) were studied bilaterally on admission for rehabilitation (21 +/- 1 days after stroke) and then at 1, 2, 3, and 6 months. Twenty healthy controls (61 +/- 5 years, 17 female) were tested once on their preferred leg. Subjects performed maximal voluntary contractions of the quadriceps and hamstring muscles. Simultaneous measurements were made of agonist force and surface EMG from agonist and antagonist muscles. Voluntary activation was estimated using the twitch superimposition technique. RESULTS: Both paretic muscles showed lower (p = 0.01 0.0005) voluntary strength than both non-paretic and control muscles until three months after stroke. Co-contraction of antagonists was similar in all groups and greater during knee extension than flexion. Stroke patients showed considerable bilateral voluntary activation failure (25-40%, p = 0.01-0.001) throughout the study while most control subjects did not (group mean 7%). CONCLUSIONS: The muscle weakness and bilateral activation failure in the stroke patients was not explained by either excessive antagonist activity or disuse atrophy. They had potential for increased voluntary strength and if this were addressed during rehabilitation, then the rate and extent of functional recovery might be enhanced. PMID- 11394589 TI - Measuring subjective quality of life following spinal cord injury: a validation study of the assistive technology device predisposition assessment. AB - PURPOSE: Assesses the validity of a subset of items of the Assistive Technology Device Predisposition Assessment (ATD PA) as a measure of quality of life (QOL) for persons with new spinal cord injury. METHOD: Subjects completed the ATD PA QOL subset, Satisfaction with Life Scale (SWLS), and Brief Symptom Inventory (BSI) while in acute rehabilitation. The internal reliability of the QOL subset of the ATD PA was assessed. Concurrent validity of the ATD PA's QOL subset with the SWLS and construct validity with the BSI depression subscale was assessed using Spearman correlations. Subjects were recruited while acute rehabilitation inpatients in a general hospital and consisted of twenty persons with newly acquired SCI (10 males and 10 females). RESULTS: Significant positive correlations between the ATD PA's QOL subset and SWLS (and significant negative correlations with the BSI depression subscale) suggest the QOL subset has concurrent and construct validity. CONCLUSIONS: The ATD PA's QOL subset appears to be a valid measure and, thus, it can be useful both in identifying subjective quality of life and predispositions to AT use early in rehabilitation. PMID- 11394590 TI - Effectiveness of two different sensory-integration programmes for children with spastic diplegic cerebral palsy. AB - PURPOSE: This study was planned to investigate the effects of individual and group approaches to sensory-perceptual-motor (SPM) training on children with cerebral palsy. METHOD: This study was carried out at the School of Physical Therapy and Rehabilitation of Hacettepe University, Occupational Therapy Unit. Forty-one children who were diagnosed as having spastic diplegic cerebral palsy by the Department of Paediatric Neurology of Hacettepe University were assessed. Forty-one children with cerebral palsy were randomly divided into three groups. Individual and group SPM training were given to first group (IND) (n = 16) and second group (GRP) (n = 16) respectively. The third group was determined as a control group (n = 9) and only the home programme was given. All children were evaluated with Ayres Southern California Sensory Integration Test and Physical Ability Test before and after training. The SPM training programmes were applied for 1.5 hours, 3 days per week for 3 months. RESULTS: Estimates of effect sizes were calculated for group, individual and control treatments. Results indicated that both group and individual treatments had a measurable effect that was consistently greater than that of controls. CONCLUSIONS: This study shows that programmes of SPM training in individuals and group treatments affect children with cerebral palsy. It was concluded that SPM training in children with cerebral palsy will be applied to combined programmes and the relationship with individual and group treatments developed. PMID- 11394591 TI - The stability of impact of assistive devices. AB - PURPOSE: Abandonment of an assistive device (AD) is a frustrating issue yet is very common. An important reason for abandonment is that the device fails to improve the quality of life of the adopter. METHOD: By means of a new questionnaire, the Psychosocial Impact of Assistive Devices Scale (PIADS) the impact on quality of life of any AD can be measured. This study tested the hypothesis that the impact of the adoption of an AD would wane with time, as the user becomes more adapted to the AD. The study attempted to do a one year follow up on all patients at a leading Canadian clinic who had received first-time prescriptions for eyeglasses. RESULTS: The results showed that the positive impact, present on adoption, did not diminish significantly for those who retained the use of the device. CONCLUSION: The PIADS provides clinicians with a reliable and economical method for assessing the role of psychosocial factors in the retention or abandonment of an AD. PMID- 11394592 TI - Tsunami. PMID- 11394593 TI - Anterior cruciate ligament reconstruction. A prospective randomized study of three surgical methods. AB - A prospective randomized study was performed to determine the differences in results between three methods of anterior cruciate ligament reconstruction: autogenous bone-patellar tendon-bone graft (group 1), semitendinosus and gracilis tendon graft reconstruction combined with an extraarticular procedure (group 2), and semitendinosus and gracilis tendon graft reconstruction alone (group 3). Preoperatively, there were no significant differences between groups. At a mean of 35.4 +/- 11.6 months postoperatively, 102 patients returned for evaluation. International Knee Documentation Committee knee evaluation revealed no significant differences in symptoms, function, return to pre-injury activity, harvest site abnormalities, or limitation of motion between groups 1 and 3. Patients in group 2 had a higher incidence of patellofemoral crepitation and loss of motion than did patients in group 3. The mean manual maximum KT-1000 arthrometer side-to-side difference was 2.1 +/- 2.0 mm in group 1, which was statistically significantly better than the difference in group 3 (3.1 +/- 2.3 mm). Final knee rating showed that 34 of 35 patients in group 1, 23 of 34 patients in group 2, and 26 of 33 patients in group 3 had a normal or nearly normal overall knee rating. Anterior cruciate ligament reconstruction with a semitendinosus and gracilis or a patellar tendon autograft may yield similar subjective results; however, the patellar tendon autograft may provide better objective stability in the long term. In addition, there appears to be no benefit to combining an intraarticular anterior cruciate ligament reconstruction with an extraarticular procedure. PMID- 11394594 TI - Fixed tibial subluxation after successful anterior cruciate ligament reconstruction. AB - Anterior cruciate ligament reconstruction results in improved function and stability in many patients. However, it is not known whether the improved stability is associated with an improved tibiofemoral relationship. We used stress radiographs to determine not only stability but also the tibiofemoral relationship in 15 patients who had a clinically successful anterior cruciate ligament reconstruction. Their results were compared with those of 14 volunteers with normal knees. The average Lysholm score for the patients was 94. None of the patients had more than 3 mm of side-to-side difference on KT-1000 arthrometer testing. Maximal anteroposterior tibial translation as measured by stress radiography was slightly increased in the reconstructed knees but was not statistically significantly different (6.6 +/- 3.2 mm versus 5.0 +/- 3.3 mm). However, with a posteriorly directed stress the tibia in the reconstructed knees did not translate posteriorly to the same extent as did the control knees, resulting in a significant difference in tibial position (-1.2 +/- 3.0 mm versus 4.0 +/- 3.3 mm). Surgical anterior cruciate ligament reconstruction may result in reduced anteroposterior tibial translation, accomplished, in part, through restraining posterior translation, leaving the tibia with persistent subluxation. Fibrosis and contracture of the posterior structures may explain this phenomenon. PMID- 11394595 TI - The axial view to measure posterior laxity of the knee. A mathematical analysis. AB - Posterior laxity of the knee can be assessed clinically, but interpretation of the amount of displacement is highly subjective. Mechanical methods are more efficacious for measuring anterior laxity. Radiologic techniques are available for measurement in a lateral projection, but some variables may interfere with their accuracy. We undertook a trigonometric analysis of the axial view to confirm that it can be used to reliably measure posterior displacement. The ideal radiologic conditions consist of 80 degrees of knee flexion in both knees maintained with a knee support and a 26 degrees x-ray beam incidence with respect to the tibia. Although such accuracy is rarely obtained in routine clinical practice, even with a variability of +/- 10 degrees in the x-ray beam incidence the error factor will be less than 10%. Such a degree of error is in the same range (+/- 2 mm) as noted by investigators using the lateral radiograph to measure anteroposterior displacement. We believe that our study validates the use of a single axial radiograph of both knees to assess the state of the posterior cruciate ligament of an injured knee as compared with a normal knee. PMID- 11394596 TI - Cryotherapy: an effective modality for decreasing intraarticular temperature after knee arthroscopy. AB - Cryotherapy is a modality commonly used after arthroscopic procedures. We divided 17 patients into two groups after routine knee arthroscopy: 12 patients were immediately treated with ice and 5 control patients were treated without ice for the first hour. In all patients, thermocouple probes were placed intraarticularly into the lateral gutter of the knee. Ice was placed on the operative knees of the treatment group for 2 hours. The control group had no intervention for the 1st hour and then had ice applied for the 2nd hour. Temperatures were continually recorded every minute for 2 hours. The temperature in the treatment group declined significantly, by 2.2 degrees C (95% confidence interval [-3.6 degrees C, -0.72 degrees C]) over the 1st hour and by 0.79 degrees C (95% CI [-1.8 degrees C, 0.18 degrees C]) over the 2nd hour (P = 0.008). The temperature in the control group increased significantly, by 5.0 degrees C (95% CI [2.4 degrees C, 7.5 degrees C]) over the 1st hour (P = 0.006). After ice was applied, the temperature fell significantly, by 4.0 degrees C (95% CI [-8.3 degrees C, 0.26 degrees C]) (P = 0.06). The difference between the temperature decrease in the treatment group and the increase in the control group at 60 minutes was 7.1 degrees C. This is the first rigorously conducted study in human patients that documents a statistically significant decline in intraarticular knee temperature with the application of ice and compression to the skin. The mechanism by which cryotherapy acts must therefore include the cooling effect on the intraarticular environment and synovium. PMID- 11394597 TI - Diagnostic performance of clinical examination and selective magnetic resonance imaging in the evaluation of intraarticular knee disorders in children and adolescents. AB - To determine the diagnostic performances of clinical examination and selective magnetic resonance imaging in the evaluation of intraarticular knee disorders in children and adolescents we compared them with arthroscopic findings in a consecutive series of pediatric patients (< or = 16 years old). Stratification effects by patient age and magnetic resonance imaging center were examined. There were 139 lesions diagnosed clinically, 128 diagnosed by magnetic resonance imaging, and 135 diagnosed arthroscopically. There was no significant difference between clinical examination and magnetic resonance imaging with respect to agreement with arthroscopic findings (clinical examination, 70.3%; magnetic resonance imaging, 73.7%), overall sensitivity (clinical examination, 71.2%; magnetic resonance imaging, 72.0%), and overall specificity (clinical examination, 91.5%; magnetic resonance imaging, 93.5%). Stratified analysis by diagnosis revealed significant differences only for sensitivity of lateral discoid meniscus (clinical examination, 88.9%; magnetic resonance imaging, 38.9%) and specificity of medial meniscal tears (clinical examination, 80.7%; magnetic resonance imaging, 92.0%). For magnetic resonance imaging, children younger than 12 years old had significantly lower overall sensitivity (61.7% versus 78.2%) and lower specificity (90.2% versus 95.5%) compared with children 12 to 16 years old. There was no significant effect of magnetic resonance imaging center. In conclusion, selective magnetic resonance imaging does not provide enhanced diagnostic utility over clinical examination, particularly in children, and should be used judiciously in cases where the clinical diagnosis is uncertain and magnetic resonance imaging input will alter the treatment plan. PMID- 11394598 TI - A comparison of ultrasonic suture welding and traditional knot tying. AB - The slippage of knots and the technical challenge of tying them securely are potential impediments to certain arthroscopic procedures. Ultrasonic energy delivered at 70 kHz can be used to weld No. 2 polypropylene suture. This method was compared with a traditional knot (surgeon's knot with four alternating half hitches) tied with an open technique to determine whether welding of sutures is comparable, in mechanical properties, to hand-tied knots. Both loops were fashioned around a 0.25-inch mandrel and then tested. The load to reach 3-mm elongation (point of likely biologic failure of a repair) was significantly greater for welded sutures than for knots. The elongation at ultimate failure was significantly less for welded sutures than for knots. The number of cycles to failure and the creep after initial displacement were similar for both welded and knotted suture loops. The ultimate load to failure was significantly greater for the knotted than for the welded suture. The welding of suture for the repair of musculoskeletal soft tissue presents an attractive alternative to traditional knot tying, particularly for arthroscopic applications. PMID- 11394599 TI - Intrinsic and extrinsic risk factors for muscle strains in Australian football. AB - Muscle strains are common injuries in Australian football and other sports involving sprinting. Between 1992 and 1999, 83,503 player-matches in the Australian Football League were analyzed for risk of muscle strain injuries using logistic regression analysis. There were 672 hamstring, 163 quadriceps, and 140 calf muscle strain injuries. All three types of muscle strains were associated with significant risk factors. For all injuries, the strongest risk factor was a recent history of that same injury and the next strongest risk factor was a past history of the same injury. History of one type of muscle strain increased the risk for certain types of other muscle strains. Age was a risk factor for hamstring and calf muscle strains (even when adjusted for injury history) but was not a risk factor for quadriceps muscle strains. Quadriceps muscle injuries were more common in shorter players and were more likely when there had been less rainfall at the match venue in the previous week. Quadriceps muscle injuries were significantly more common in the dominant kicking leg, whereas hamstring and calf muscle injuries showed no difference in frequency between the dominant and nondominant legs. PMID- 11394600 TI - Risk factors for recurrent stress fractures in athletes. AB - Our aim was to identify factors predisposing athletes to multiple stress fractures, with the emphasis on biomechanical factors. Our hypothesis was that certain anatomic factors of the ankle are associated with risk of multiple stress fractures of the lower extremities in athletes. Thirty-one athletes (19 men and 12 women) with at least three separate stress fractures each, and a control group of 15 athletes without fractures completed a questionnaire focusing on putative risk factors for stress fractures, such as nutrition, training history, and hormonal history in women. Bone mineral density was measured by dual-energy x-ray absorptiometry in the lumbar spine and proximal femur. Biomechanical features such as foot structure, pronation and supination of the ankle, dorsiflexion of the ankle, forefoot varus and valgus, leg-length inequality, range of hip rotation, simple and choice reaction times, and balance in standing were measured. There was an average of 3.7 (range, 3 to 6) fractures in each athlete, totaling 114 fractures. The fracture site was the tibia or fibula in 70% of the fractures in men and the foot and ankle in 50% of the fractures in women. Most of the patients were runners (61%); the mean weekly running mileage was 117 km. Biomechanical factors associated with multiple stress fractures were high longitudinal arch of the foot, leg-length inequality, and excessive forefoot varus. Nearly half of the female patients (40%) reported menstrual irregularities. Runners with high weekly training mileage were found to be at risk of recurrent stress fractures of the lower extremities. PMID- 11394601 TI - The effect of exercise on anterior-posterior translation of the normal knee and knees with deficient or reconstructed anterior cruciate ligaments. AB - Exercise may result in increased laxity in the knee. Anterior translation in 40 normal knees, 33 consecutive anterior cruciate ligament-deficient knees, and 30 randomly chosen anterior cruciate ligament-reconstructed knees was measured using the KT-1000 arthrometer before and after the participants ran for 15 minutes on a neutral-incline treadmill. A single observer blinded to the status of each knee tested all participants. There was a significant increase in anterior translation in the normal (mean, 0.75 mm), anterior cruciate ligament-deficient (mean, 0.62 mm), and anterior cruciate ligament-reconstructed knees (mean, 0.25 mm) after exercise. In addition, the amount of anterior translation after exercise was significantly different when these groups were compared with each other. Post hoc analysis using Tukey's procedure indicated that anterior translation in the anterior cruciate ligament-reconstructed knee was significantly less than in the normal and anterior cruciate ligament-deficient knees. Therefore, repetitive loading exercise contributes to an increase in anterior translation in normal, anterior cruciate ligament-deficient, and anterior cruciate ligament reconstructed knees, and the anterior cruciate ligament-reconstructed knee does not respond to repetitive loading in the same manner as a normal knee. PMID- 11394602 TI - Outcome of surgery for chronic Achilles tendinopathy. A critical review. AB - Achilles tendinopathy is often treated surgically after failure of nonoperative management, but results are not uniformly excellent. We critically assessed the methods of 26 studies that reported surgical outcomes of patients with this condition. Using 10 previously published criteria, and blinded to study outcomes, we derived a "methodology score" (0 to 100) for each study. This score was highly reproducible (r = 0.99, P < 0.01). Scores were generally low concerning the type of study, subject selection process, and outcome measures, which indicates methods deficiency in the way the study was designed, performed, and analyzed. We found a negative correlation between reported success rate and overall methods scores (r = -0.53, P < 0.01), and a positive correlation between year of publication and overall methods score (r = 0.70, P < 0.01). Study methods may influence reported surgical outcome, and we suggest guidelines for improving study design in this area of clinical research. We acknowledge that study methods have improved over the course of the past 20 years. PMID- 11394603 TI - Biomechanical behavior of the plantar flexor muscle-tendon unit after an Achilles tendon rupture. AB - The purpose of this study was to examine the biomechanical behavior of the plantar flexor muscle-tendon unit in subjects who had ruptured their Achilles tendon. Twenty-six men and 14 women volunteered for the study. Eighteen subjects had been treated operatively and 22, nonoperatively. All subjects had ruptured their Achilles tendon more than 1 year before the study, and 28 of the 40 ruptures occurred 5 years or less before the day of testing. A KinCom dynamometer was used to measure ankle joint angle, passive torque, and maximal isometric plantar flexor torque. During a 2-minute passive calf stretch, stiffness and torque relaxation were calculated. Isometric torque and peak passive torque were 17% and 10% greater for the uninvolved versus the involved limb, whereas stiffness and torque relaxation were not different between limbs. The time since injury did not influence the results, nor did the mode of initial treatment, that is, whether the subjects were treated operatively or nonoperatively. These findings suggest that changes in strength and peak passive torque may be chronic adaptations associated with Achilles tendon rupture. PMID- 11394604 TI - Does intraarticular morphine improve pain control with femoral nerve block after anterior cruciate ligament reconstruction? AB - In a prospective, randomized, double-blinded manner, we compared the effects of a preoperative intraarticular injection of morphine (5 mg) or a placebo, combined with a postoperative femoral nerve block, on postoperative pain. Sixty-two patients underwent an arthroscopically assisted anterior cruciate ligament reconstruction using patellar tendon autograft under general anesthesia. No statistical difference between the two groups was evident in terms of age, sex, weight, operative time, volume of bupivacaine received with the femoral nerve block, or tourniquet use or tourniquet time. Comparison of visual analog scale pain scores revealed no statistical difference between the groups at any point after the operation. Both groups had a significant decrease in visual analog scale scores after the femoral nerve block, with the lowest mean values 4 hours after the operation (morphine group, 1.7; placebo group, 1.4), and continuing to be significantly less through 24 hours (morphine, 2.6; placebo, 2.9). No significant difference in postoperative narcotic medication use was evident in the recovery room or at home. A post hoc power analysis revealed that the study power reached 87%, with a significance level of 5%. The postoperative femoral nerve block was effective, and intraarticular morphine provided no additional benefit. PMID- 11394605 TI - Snowboarder's talus fracture. Mechanism of injury. AB - Fracture of the lateral process of the talus is an injury unique to snowboarders and is of particular clinical relevance because it masquerades as an anterolateral ankle sprain and is difficult to detect on standard radiographic views. Misdiagnosis can lead to long-term morbidity in a young and active population, with ensuing severe degeneration of the subtalar joint. To date, the precise mechanism of injury has not been established, making it difficult to identify potential preventive strategies in equipment design or snowboarding technique. Fracture of the lateral process of the talus in snowboarders has been thought to result from pure dorsiflexion and inversion combined with axial loading. We hypothesized, however, that external rotation is a key component of the mechanism of injury. Ten cadaveric ankles were mounted on a materials testing machine in a position of fixed dorsiflexion and inversion. All ankles were loaded to failure axially, with or without combined external rotation. No fractures occurred after axial loading in dorsiflexion and inversion, but six of eight specimens sustained fractures of the lateral process of the talus when similarly loaded with external rotation added, supporting our hypothesis. Further study is needed to evaluate the relationship between various types of snowboarding equipment and fracture mechanism. PMID- 11394606 TI - Anterior cruciate ligament function after tibial eminence fracture in skeletally mature patients. AB - We compared anterior cruciate ligament function in skeletally mature patients after treatment of tibial eminence fractures with that of patients in two other groups: patients who had anterior cruciate ligament deficiency and patients who had undergone anterior cruciate ligament reconstruction using bone-patellar tendon-bone autografts. The Lysholm questionnaire was used to evaluate symptoms and KT-1000 arthrometry was used to determine objective knee laxity at an average follow-up of 5.2 years. Knee joint proprioception was assessed with a new method designed to test joint position sense. The Lysholm score for the tibial eminence group was 94 +/- 7. Only the patients in the anterior cruciate ligament-deficient group demonstrated statistically significantly increased laxity and inferior proprioception when the injured leg was compared with the uninjured leg. Both laxity and proprioception were statistically inferior for the anterior cruciate ligament-deficient group when compared with both the treated tibial eminence fracture group and the anterior cruciate ligament-reconstructed group. No statistically significant difference was observed between the anterior cruciate ligament-reconstructed and treated tibial eminence fracture groups. Correlation was observed between laxity and proprioception when all patients were analyzed. The results demonstrate that appropriate treatment of tibial eminence fractures restores stability and proprioception to the knee. PMID- 11394607 TI - Medially based anterior capsular shift of the glenohumeral joint. Passive range of motion and posterior capsular strain. AB - The effect of a medially based anterior capsular shift on translational and rotational range of motion and posterior capsular strain was investigated in an in vitro model. Six cadaveric shoulders were tested in a robot-assisted shoulder simulator. Translational and rotational range of motion were reduced by the capsular shift, particularly with the shoulder at higher elevation angles. At 60 degrees of elevation, anterior translation was decreased 1.9 +/- 2.9 mm, and posterior translation was decreased 2.3 +/- 6.2 mm. External rotation was decreased 11.5 degrees +/- 10.2 degrees, and internal rotation was decreased 8.9 degrees + 5.7 degrees. Posterior capsular strain change was dependent on elevation angle. At 30 degrees of elevation, strain tended to increase 5.0% +/- 7.3% in the inferior aspect and 2.9% +/- 2.6% in the superior aspect, with no change detected in the medial aspect. At 60 degrees of elevation, strain increased 6.6% +/- 8.0%, 3.2% +/- 2.6%, and 4.4% +/- 3.5% in the inferior, middle, and superior aspects, respectively. Our results support the use of the medially based anterior shift for shoulders with anterior-inferior instability or multidirectional instability with posterior involvement. PMID- 11394608 TI - Relationships between throwing mechanics and shoulder distraction in professional baseball pitchers. AB - The extreme forces and torques and the high speeds and excessive ranges of motion of baseball pitching place tremendous stress on the soft tissues of the throwing shoulder. Little is known about the relationship between pitching mechanics and shoulder joint stress, especially in professional athletes. The purpose of this study was to quantify joint loads and kinematic parameters of pitching mechanics at the major league level and to study their relationships. Three-dimensional, high-speed video data were collected on 40 professional pitchers during the 1998 Cactus League spring training. A clinically significant distraction force was calculated at the shoulder joint, which reached an average peak value of 947 +/- 162 N (108% +/- 16% body weight). Descriptive statistics and a multiple linear regression analysis were used to relate shoulder distraction to kinematic and kinetic parameters of pitching mechanics. This study was undertaken not only to investigate the peak forces and torques on the shoulder, but also to identify potential areas of intervention that might prevent throwing injuries. Knowledge of joint ranges of motion, angular velocities, and joint-reaction forces can provide a scientific basis for improved preventive and rehabilitative protocols for baseball pitchers. PMID- 11394609 TI - Postural control after anterior cruciate ligament reconstruction and functional rehabilitation. AB - Total sagittal knee laxity and postural control in the sagittal and frontal planes were measured in 25 patients at a mean of 36 months (range, 27 to 44) after anterior cruciate ligament reconstruction and in a control group consisting of 20 uninjured age- and activity-matched subjects. Body sway was measured in the sagittal plane on a stable and on a sway-referenced force plate in single-legged stance, double-legged stance, or both, with the eyes open and closed. Postural reactions to perturbations in the sagittal and frontal planes were recorded in the single-legged stance with the eyes open. Total sagittal plane laxity was significantly greater in the anterior cruciate ligament-reconstructed knee (11.2 mm; range, 6 to 15) than in the uninjured knee (8.9 mm; range, 6 to 12) or in the control group (6.0 mm; range, 5 to 8). In spite of this, the patients, in comparison with the controls, exhibited normal postural control except in two variables-the reaction time and the latency between the start of force movement to maximal sway in the sagittal plane perturbations. This supports the hypothesis that rehabilitation, with proprioceptive and agility training, is an important component in restoring the functional stability in the anterior cruciate ligament reconstructed knee. PMID- 11394610 TI - Synovial sarcoma diagnosed after a sports injury. PMID- 11394611 TI - Stress fracture of the clavicle in a female lightweight rower. A case report and review of the literature. PMID- 11394612 TI - Peroneus quartus muscle. A rare cause of chronic lateral ankle pain. PMID- 11394613 TI - Athletic activity after joint replacement. AB - The first decade of the 21st century has been declared the "Bone and Joint Decade" by 35 nations and 44 states in the United States as of March 2001. It is not surprising that Americans are interested in musculoskeletal disease and the treatment of bone and joint disorders because our population is aging, the prevalence of arthritic joints is increasing, and senior Americans are demonstrating a strong desire to stay active in activities of daily living and athletics. One of the most successful treatments for painful arthritic joints, which limit activity, is total joint replacement, which predictably relieves pain and improves function. Much has been written about the technical aspects of total joint arthroplasty. Less has been written about safe and appropriate activities for patients who have had joint replacement operations. This article evaluates athletic activity after joint replacement by reviewing the orthopaedic literature and surveying members of The Hip Society, The Knee Society, and The American Shoulder and Elbow Surgeons Society. The authors have developed consensus recommendations for appropriate athletic activity for patients who have had joint replacement operations. This article is intended to serve as a guide for orthopaedic surgeons and primary care physicians who give patients recommendations for athletic activity after joint replacement. This article is also intended to stimulate further research in the area of athletic activity after total joint arthroplasty. PMID- 11394614 TI - Injuries to the shoulder in the throwing athlete. Part two: evaluation/treatment. PMID- 11394615 TI - Immediate surgical repair of the medial patellar stabilizers for acute patellar dislocation. A review of eight cases. PMID- 11394616 TI - Voluntarily evoked positive Lachman test produced by gastrocnemius muscle contraction. A report of three cases. PMID- 11394617 TI - Men with osteoporosis: are they being treated fairly? PMID- 11394618 TI - Do osteoporotic vertebral deformities deserve medical attention? PMID- 11394619 TI - Surgical treatment of the rheumatoid elbow. AB - In rheumatoid arthritis, the elbow is involved in 20 to 50% of the cases. Surgical treatment for rheumatoid arthritis is proposed to patients in whom an appropriate and adequate attempt at medical management has failed. Improvements in surgical technique and prosthetic design have led to more predictable results in the surgical treatment of the rheumatoid elbow. Surgical treatment of the patient with rheumatoid arthritis continues to evolve. Synovectomy continues to be an effective palliative procedure, preferred in the early stages of disease (I, II, IIIA) with or without radial head resection. Further investigation into the use of arthroscopic techniques may result in decreased morbidity and a quicker recovery. In more advanced stages (IIIA, IIIB, IV), total elbow replacements by experienced surgeons employing contemporary designs, unconstrained or semiconstrained, and surgical techniques are associated with a high degree of success with long-term follow-up, approaching that fortotal hip and knee replacement. Finally, interposition arthroplasty can be proposed for young adults with stage II or IIIA rheumatoid arthritis in whom the elbow is mainly stiff and painful. PMID- 11394620 TI - Phosphate, the renal tubule, and the musculoskeletal system. AB - A component of ATP, phosphate is at the hub of the energy-related mechanisms operative in muscle cells. Together with calcium, phosphate is involved in bone tissue mineralization: thus, a chronic alteration in the metabolism of phosphate can induce bone and joint disorders. Diagnosis of chronic hypophosphatemia. Serum phosphate, calcium, and creatinine should be assayed simultaneously. Serum calcium is increased in hypophosphatemia caused by hyperparathyroidism and decreased in osteomalacia. Urinary phosphate excretion should be measured in patients with a normal serum calcium level and a serum phosphate level lower than 0.80 mmol/L. A decrease in urinary phosphate excretion to less than 10 mmol/24 h strongly suggests a gastrointestinal disorder, such as malabsorption, antacid use, or chronic alcohol abuse. In patients with a urinary phosphate excretion greater than 20 mmol/24 h, the maximal rate of tubular reabsorption of phosphate (TmPO4) and the ratio of TmPO4 over glomerular filtration rate (GFR) should be determined to look for phosphate diabetes. Manifestations and causes of phosphate diabetes in adults. Moderately severe phosphate diabetes in adults manifests as chronic fatigue, depression, spinal pain, and polyarthralgia, with osteoporosis ascribable to increased bone resorption. Although many cases are idiopathic, investigations should be done to look for X-linked vitamin D-resistant rickets missed during childhood, a mesenchymatous tumor, or Fanconi's syndrome with renal wasting of phosphate, glucose, and amino acids. Management of phosphate diabetes. Phosphate supplementation and, in patients with normal urinary calcium excretion, calcitriol produce some improvement in the symptoms and increase the bone mineral density. Whether dipyramidole is clinically effective remains unclear. PMID- 11394621 TI - Should percutaneous vertebroplasty be used to treat osteoporotic fractures? An update. AB - Acrylic cement vertebroplasty is being increasingly used to treat osteoporotic vertebral compression fractures (VCFs), although no controlled studies supporting this trend have been published. Vertebroplasty remains controversial as a treatment for osteoporotic fractures because it is a local response to a systemic disease and because the pain caused by osteoporotic fractures usually subsides within a few days or weeks. Current data suggest that pain severity may decrease by half, on average, in 90-100% of patients. Although vertebroplasty is usually well tolerated, serious neurological complications have been reported in a few patients. The most common adverse event is nerve root pain, usually caused by leakage of the cement into the intervertebral foramen. Whether vertebroplasty is followed by an increased risk of osteoporotic fractures in the adjacent vertebras remains unclear. Resorbable cements are being developed and may provide better results than the acrylic cements used today. At present, acrylic cement vertebroplasty to treat osteoporotic VCFs is appropriate in only a minority of patients selected carefully by a multidisciplinary team including a rheumatologist. PMID- 11394622 TI - Ultrasound versus MRI in the evaluation of juvenile idiopathic arthritis of the knee. AB - OBJECTIVE: To examine the role of ultrasound versus magnetic resonance imaging (MRI) in assessing joint inflammation in patients with juvenile idiopathic arthritis (JIA) of the knee. METHODS: This study was conducted on 38 patients with juvenile idiopathic arthritis (25 girls and 13 boys), whose ages ranged between 2-17 years (mean 8 years), presenting with joint swelling, tenderness, pain on motion and/or limitation of movement. Plain radiography, high-resolution ultrasound and MRI examinations of the knee (before and after contrast administration) were made on all patients. A control group of ten subjects was also examined. RESULTS: Compared to the control group, sonographic examination was found to be of great value as regards the joint effusion, popliteal cysts, lymph nodes and to a lesser extent, the degree of affection of the articular cartilage. MRI was superior in evaluating the extent of synovial proliferation (pannus), thinning out and erosions of articular cartilage, loculated effusions as well as hypoplastic menisci and ligaments, especially after contrast enhancement. CONCLUSION: Ultrasound is a simple, inexpensive and valuable tool in evaluating the initial stages of JIA. In more advanced stages of JIA and also for monitoring the progression of the disease process and response to therapy, MRI examination following gadolinium proved to be superior in evaluation of the joint affection. PMID- 11394623 TI - EULAR recommendations for the management of knee osteoarthritis. Report of a task force of the Standing Committee for International Clinical Studies Including Therapeutic Trials. AB - A task force for the EULAR standing committee for clinical trials determined the methodological and logistical approach required for the development of evidence based guidelines for treatment of knee osteoarthirits (OA). METHODS: The first stage was the selection of treatment modalities to be considered. The second stage comprised a search of the databases of all European-language publications. All of the relevant studies were quality scored. The third stage involved determination of key clinical propositions by expert consensus employing a Delphi approach. The final stage involved ranking of these propositions according to the available evidence. A second set of propositions relating to a future research agenda was determined by expert consensus. RESULTS: Seven hundred and forty-four studies presented outcome data of the effects of specific treatments on knee OA. Quantitative analysis of treatment effect was possible in only 61 studies. Recommendations for the management of knee OA based on currently available data and expert opinion are presented. Proposals for a future research agenda are highlighted. CONCLUSIONS: These are the first clinical guidelines on knee OA to combine an evidence-based approach and a consensus approach across a wide range of treatment modalities. It is apparent that only certain clinical propositions are supported by substantial research-based evidence. There is thus an urgent need for future well-designed trials to address key clinical questions. PMID- 11394624 TI - Shockwave therapy under ultrasonographic guidance in rotator cuff calcific tendinitis. AB - OBJECTIVES: This is the first efficacy evaluation in patients with rotator cuff calcific tendinitis treated with shockwave therapy delivered using a dedicated extracorporeal lithotripter and ultrasonographic guidance. PATIENTS AND METHODS: Thirty-two patients with rotator cuff calcific tendinitis received shockwave therapy delivered using a lithotripter specifically designed for the musculoskeletal system (DORNIER EPOS ULTRA), under ultrasonography guidance (which is not part of the conventional procedure). The cumulative dose per patient was 1300 mJ/mm2 divided in two or three sessions. Three, six, 12, and 24 weeks after treatment, the patients were evaluated using a visual analog scale for pain, the ASES self-questionnaire, Constant's score, and plain radiographs. RESULTS: Improvements in the pain and self-questionnaire scores were noted in 36.6% of patients after 12 weeks and 55.1% after 24 weeks. Plain radiographs were changed in 26.6% of patients after 12 weeks (with complete clearance of the calcific deposits in 6.6%) and in 24.1% of patients after 24 weeks (complete clearance in 17.2%). CONCLUSION: These results seem less favorable than those reported with dedicated lithotripters and computed tomography guidance. PMID- 11394625 TI - Chronic recurrent multifocal osteomyelitis: five-year outcomes in 14 pediatric cases. AB - OBJECTIVE: To determine the clinical presentation and outcomes of chronic recurrent multifocal osteomyelitis (CRMO) in pediatric patients. PATIENTS AND METHODS: Ten girls and four boys were followed up between 1993 and 1999 for CRMO diagnosed on the basis of radiographic bone lesions with, at the same sites, increased radionuclide uptake, negative microbiological specimens, and histological evidence of nonspecific osteomyelitis. RESULTS: Mean age at CRMO was 9.6+/-3.4 years, mean disease duration was 5.3+/-2.5 years, and mean number of flares per patient was 5.9+/-3.7. Thirty-four percent of lesions were in the metaphyses of the lower limb bones, 14% in the pelvis, and 13% in the chest wall (with clavicular lesions in four patients). Three patients had skin lesions (psoriasis in two and palmoplantar pustulosis in one). Eight patients received antibiotic therapy, for 2 months at the most, to no advantage in the short term. Nonsteroidal anti-inflammatory drugs were used in all 14 patients and glucocorticoid therapy in four. Sulfasalazine was used in five patients, to good effect in four. Mean follow-up was 5.3+/-2.5 years. At last follow-up, eight patients had active disease, including one with synovitis and one with Takayashu's disease. CONCLUSION: As compared to SAPHO syndrome, skin lesions and chest wall involvement are less common in CRMO. The long-term prognosis is guarded: in our study only six of 14 patients were in remission at last follow up. PMID- 11394626 TI - Management of male osteoporosis. AB - The objective of this study was to evaluate the efficacy of treatments for male osteoporosis selected based on the cause of the disease. METHODS: Sixty-three men with osteoporosis (T-score at the lumbar spine and/or femoral neck lower than 2.5) with a mean age of 53+/-11 years were studied. Forty-three (68.3%) had a history of fracturing without trauma (vertebral fractures, 37 patients, 57%). Treatments were as follows: idiopathic osteoporosis: calcium and vitamin D supplements (N = 10) or cyclical etidronate for 2 weeks followed by calcium and vitamin D supplements for 76 days (N = 29); moderate idiopathic phosphate diabetes: calcitriol and phosphate (N = 15); idiopathic hypercalciuria: hydrochlorothiazide (N = 6); and hypogonadism: testosterone (N = 3). RESULTS: Percentage change in bone mineral density (mean +/- standard error of the mean) after 18 months: calcium and vitamin D (lumbar spine: 0.6+/-2; femoral neck: 2.2+/-2.2); etidronate (lumbar spine: 3.6+/-1.4*; femoral neck: 0.5+/-1); calcitriol (lumbar spine: 7.0+/-3.5*; femoral neck: 0.0+/-1.4); thiazide diuretic (lumbar spine: 1+/-3.2; femoral neck: -2.3+/-3.7); and testosterone (lumbar spine: 6.8+/-6.4; femoral neck: 2.5+/-2.7), where *P < 0.05 versus baseline. Gastrointestinal side effects occurred in three patients (4.8%), including two on calcitriol-phosphate therapy and one on etidronate therapy. Of the six (9.5%) patients who experienced incident fractures, four were on etidronate, one on calcitriol-phosphate, and one on calcium-vitamin D. No patients discontinued their treatment because of side effects. CONCLUSION: Etidronate and the combination of calcitriol-phosphate produce a significant increase in lumbar spine bone mass in men with idiopathic osteoporosis or moderate idiopathic phosphate diabetes. PMID- 11394627 TI - Atypical familial Paget's disease of bone. AB - We report unusual clinical and radiological features of Paget's disease of bone in three family members. All three patients had satisfactory biochemical and symptomatic response to treatment with an intravenous bisphosphonate (clodronate). PMID- 11394628 TI - Relapse of Wegener's granulomatosis. Concerning a case after 20 years of remission. AB - Immunosuppressive drugs have transformed the prognosis of systemic Wegener's granulomatosis. Nowadays, the main residual problem is illness relapses, for which management is largely undefined. We describe the case of a patient, aged 47 in 1977. The diagnosis of Wegener's granulomatosis was made when faced with polyarthralgias, cutaneous vasculitis, rhinitis, dyspnea, hemoptysis and global decline of her physical condition. The treatment associated high-dose corticotherapy and intramuscular cyclophosphamide for 1 year. This treatment led to a complete remission. Twenty years later, the patient was hospitalized for reoccurrence of rhinitis, dyspnea and right knee effusion associated with biological inflammatory syndrome, renal insufficiency and antibodies against polymorphonuclear neutrophil cytoplasm, type c-ANCA. Chest CT-scan disclosed parenchymal infiltrates. Wegener relapse was diagnosed and the combination of three methylprednisolone perfusions followed by oral prednisone (1 mg/kg/d) and a monthly bolus of cyclophosphamide led to a new remission. Nevertheless, 4 months after beginning the treatment the patient died from an infectious complication (Pneumocystis carinii and aspergillosis). Relapses of Wegener's granulomatosis are frequent and difficult to predict. Moreover, some cases occur very early. The remarkable efficiency of cyclophosphamide to induce remission is however shaded by the high rate of relapse. Other drugs are studied to identify more efficient therapy, able to both induce remission and prevent relapses, but reliable data are still missing to determine the best therapeutic regimen. PMID- 11394629 TI - Cholesterol crystal embolization simulating focal myositis. AB - Cholesterol crystal embolization usually produces characteristic skin lesions. We report a case responsible for myositis of the calf without suggestive skin lesions. The outcome in this 58-year-old patient was spontaneously favorable. Cholesterol crystal embolization can produce a range of clinical symptoms, with the skin, kidneys, and eyes being the most common targets. Generalized forms can result in systemic disease. The diagnosis rests on histological findings, and the treatment is symptomatic. Anticoagulants have been shown to worsen the manifestations, whereas antiplatelet therapy may be useful. PMID- 11394630 TI - Role of chaperones in nuclear translocation and transactivation of steroid receptors. AB - Molecular chaperones assist proteins to reach their mature and functional conformation. It has become apparent in recent years that chaperones function as part of a multiprotein heterocomplex that is potentially involved not only in protein folding, but also in intracellular trafficking and in targeting proteins for degradation. In the case of steroid receptors, the activity of the chaperone heterocomplex, as well as the proteins comprising the heterocomplex, has an effect on the observed ligand-dependent transcriptional activity of the receptor. The direct interaction between chaperones and steroid receptors makes them potential therapeutic targets in a number of pathologic conditions. In the case of cancers with steroid receptor involvement, such as breast and prostate cancer, the inhibition of chaperone activity may inhibit tumor cell growth. Conversely, enhancement of chaperone activity may be beneficial in disorders of protein misfolding, as in the case of androgen receptor aggregates found in Spinal and Bulbar Muscular Atrophy. PMID- 11394632 TI - Prolonged, symptomatic hypocalcemia with pamidronate administration and subclinical hypoparathyroidism. AB - A 62-yr-old woman with thyroid carcinoma metastatic to bone, and a history of subclinical hypoparathyroidism was admitted to the hospital in hypocalcemic crisis 5 wk after receiving iv pamidronate. The patient had tetany and laryngospasm. An electrocardiogram showed junctional rhythm with QT segment prolongation. The patient had previously maintained a low-normal serum calcium on 500-750 mg of calcium carbonate and 600 IU of vitamin D daily. One week after pamidronate administration the patient's calcium and vitamin D supplementation were inadvertently discontinued. She continued to take daily intranasal calcitonin. At the time of her hospitalization for hypocalcemia, the patient's serum calcium was 4.3 mg/dL. The patient received aggressive calcium and vitamin D supplementation. However, her serum calcium remained below 6 mg/dL for a 2-wk period, and took another week to return to the normal range. In this article, we discuss the counterregulatory responses necessary to maintain calcium homeostasis following osteoclast inhibition by bisphosphonates. We also review the risk factors for hypocalcemia following bisphosphonate administration. Pamidronate and other bisphosphonates are becoming an integral part of the management of normocalcemic patients with malignant bone disease. Therefore, awareness of risk factors for hypocalcemia and familiarity with avenues available for protection from potentially catastrophic hypocalcemia are both crucial. PMID- 11394631 TI - Endocrine paraneoplastic syndromes with special reference to the elderly. PMID- 11394633 TI - Receptor phosphorylation mediates estradiol reduction of alpha2-adrenoceptor coupling to G protein in the hypothalamus of female rats. AB - Estrogen increases evoked norepinephrine release in the hypothalamus of female rodents, in part by reducing the ability of alpha2-adrenoceptors to act as negative feed-back inhibitors of norepinephrine release. Estrogen enhancement of norepinephrine release in the hypothalamus correlates with decreased coupling of the alpha2-adrenoceptor to G protein. To determine the mechanism by which estrogen uncouples alpha2-adrenoceptors from G protein, we tested the hypothesis that estrogen increases alpha2-adrenoceptor phosphorylation. Short-term activation of endogenous serine/threonine phosphatases with protamine or treatment with exogenous phosphatase restored alpha2-adrenoceptor coupling to G protein to control levels in hypothalami from estrogen-exposed female rats. Additional experiments examined whether estrogen alters G protein-coupled receptor kinase expression or activity or serine/threonine phosphatase activity. These proteins are involved in G protein-coupled receptor phosphorylation, internalization, and recycling. Estrogen exposure reduced G protein-coupled receptor kinase mRNA, protein, and activity in the hypothalamus. Furthermore, estrogen treatment reduced serine/threonine phosphatase activity in the hypothalamus. Analysis of ligand binding in subcellular fractions demonstrated that estrogen decreases the fraction of internalized alpha2-adrenoceptors in the hypothalamus.Therefore, estrogen promotes norepinephrine release in the hypothalamus by stabilizing alpha2-adrenoceptor phosphorylation, uncoupling the receptor from G protein. Estrogen may stabilize alpha2-adrenoceptor phosphorylation by inhibiting receptor internalization and dephosphorylation. PMID- 11394634 TI - Novel demonstration of a physiologic/pharmacologic role of insulin-like growth factor-1 in ovulation in rats and action on cumulus oophorus. AB - Gonadotropins are well known to be the most important stimulus for ovarian follicular development. More recently, there is indirect evidence that insulin like growth factor-1 (IGF-1) is also a very important autocrine/paracrine factor in folliculogenesis. We had access to an analog of IGF-1, LR3IGF-1, which binds very poorly to IGF-1 binding proteins and therefore was shown by previous investigators to have biologic effects. We studied rats that were superovulated with the use of gonadotropins. We showed that the addition of LR3IGF-1 by infusion further increased the ovulation rate (statistically significant) and increased the ovarian weight in two of three strains of rats. We demonstrated that infusion of LR3IGF-1 or injection of equine chorionic gonadotropin or a combination of these two were associated with oocytes with reduced number of cumulus cells (statistically significant). We conducted an experiment to determine whether in vitro culturing with varying dosage of IGF-1 may stimulate cumulus cell replication to improve the quality of oocyte cumulus complex. IGF-1 did not show any change in this respect. Through these physiologic (pharmacologic) studies, we have shown that IGF-1 (analog) can further increase the ovulation rate induced by gonadotropins. PMID- 11394635 TI - Persistent hormonal effects of stress are not due to reduced food intake or exposure to stressed rats. AB - Exposure to inescapable stress elicits persistent effects on the physiology and behavior of rats. Elevated basal plasma corticosterone concentrations have been observed for several days after cessation of stress. In this study, we measured hormonal concentrations in multiple axes at multiple levels, 24 h after one or three consecutive exposures to the same stress paradigm. The data indicated persistent activation of plasma corticosterone and prolactin concentrations, whereas plasma triiodothyronine, thyroxine, luteinizing hormone, and growth hormone concentrations were inhibited after either one or three stress sessions. In addition, we isolated the effects of restraint/tail shock per se from the effects of being moved and exposed to other stressed rats, and from the effects of reduced feeding produced by our stress protocol. The data clearly indicated that the stress paradigm, rather than exposure to stressed rats or decreased nutrient intake, is necessary to induce the persistent physiologic changes we observe after stressor exposures. PMID- 11394636 TI - GABA inhibition of immortalized gonadotropin-releasing hormone neuronal excitability involves GABA(A) receptors negatively coupled to cyclic adenosine monophosphate formation. AB - Gamma-aminobutyric acid (GABA) has been implicated in the regulation of reproduction, particularly in the developmental modulation of gonadotropin releasing hormone (GnRH) secretion. GnRH neurons are innervated by GABA containing processes, and the administration of GABA stimulates and inhibits GnRH secretion in vivo and in vitro. We have previously shown that GABA can exert both of these actions in sequence, by acting directly on immortalized GnRH neurons. While the stimulation is the result of a GABA(A) receptor-mediated depolarization of the plasma membrane, the mechanism involved in the delayed inhibition is the subject of the present investigation. GABA (1 nM-10 microM) decreased the intracellular concentration of cyclic adenosine monophosphate (cAMP) in a dose- and time-dependent fashion. This effect was blocked by bicuculline and mimicked by muscimol but not by baclofen. To analyze the effect of GABA on cellular excitability, we used fura-2 loaded GT1-7 cells. Activation of voltage-sensitive calcium channels by high K+-induced depolarization (35 mM) increased [Ca2+]i. GABA (10 microM) and muscimol (10 microM) reduced the amplitude of K+-induced [Ca2+]i transients. This inhibition was blocked by forskolin (20 microM) or 8-Br cAMP (1 mM). Altogether, these results show that GABA(A) receptors mediate a sustained inhibitory effect of GABA on GnRH neurons, and suggest the involvement of the cAMP pathway decreasing cellular excitability. PMID- 11394637 TI - Tissue kallikrein in human placenta in early and late gestation. AB - This study was addressed to identify kallikrein mRNA and protein in early, preterm, and term human placenta and to evaluate their temporospatial pattern. Kallikrein mRNA was expressed in syncytio/cytotrophoblasts and in the endothelial cells of the floating villi, with a greater intensity in early samples (isolated spontaneous abortions and ectopic pregnancies). Cytotrophoblasts at the base of the anchoring villi, maternal decidua and decidual arteries, endothelial cells of chorionic and basal plate blood vessels, and the amniotic epithelium presented a positive signal. Tissue kallikrein was predominantly observed in syncytiotrophoblasts and had a greater immunoreactivity in first-trimester samples. Intraarterial trophoblasts, blood vessels of the floating villi, basal and chorionic plates, and the amniotic epithelium showed positive immunoreactivity. The sites and variations of the tissue kallikrein mRNA and protein in the human placenta, in different stages of pregnancy, support the hypothesis that this enzyme may participate in the establishment and maintenance of placental blood flow through vasodilation, platelet antiaggregation, cell proliferation, and trophoblast invasion. PMID- 11394638 TI - High-level bacterial expression of a natively folded, soluble extracellular domain fusion protein of the human luteinizing hormone/chorionic gonadotropin receptor in the cytoplasm of Escherichia coli. AB - We have expressed the extracellular domain of the human luteinizing hormone/chorionic gonadotropin (hLH/CG) receptor as a fusion protein with thioredoxin in the cytoplasm of an Escherichia coli strain that contains mutations in both the thioredoxin reductase and glutathione reductase genes. The chimeric protein isolated following induction of expression is purified in a soluble form and binds hCG with an affinity approximating that of native receptor. This truncated form of the receptor displays the same specificity as intact hLH/CG receptor and does not bind human follicle stimulating hormone. This cytoplasmically produced protein is expressed at levels that exceed 10 mg/L. Expression of properly folded extracellular domain of the hLH/CG receptor in the cytoplasm of E. coli allows the facile and economic purification of large quantities of material. This will facilitate the determination of the structure of the hormone-binding domain of the glycoprotein receptor as well as the production of epitope-specific antibodies. PMID- 11394640 TI - Effects of endomorphin-1 on open-field behavior and on the hypothalamic-pituitary adrenal system. AB - The effects of endomorphin-1 (EM1) on behavioral responses and on the hypothalamic-pituitary-adrenal system were investigated in mice. Locomotor activity was measured in an "open-field" apparatus, with parallel recording of the numbers of rearings and groomings. Different doses of the peptide (250 ng to 5 microg) were administered to the animals intracerebroventricularly 30 min before the tests. EM1 caused significant increases in the locomotor activity and the number of rearings. The effect of EM1 on the basal corticosterone secretion was also investigated. At a dose of 5 microg, the peptide significantly increased plasma corticosterone level. The corticotropin-releasing hormone (CRH) antagonist alpha-helical CRH9-41, applied 30 min prior to EM1 administration, completely abolished the increases in both locomotion and the number of rearings and attenuated the corticosterone release evoked by EM1. These results suggest that the EM1 -induced increases in locomotion and rearing activity as well as the pituitary-adrenal activation are mediated by CRH. PMID- 11394639 TI - Peripubertal paternal EtOH exposure. AB - Fetal alcohol syndrome usually implies effects on the offspring of maternal EtOH consumption during gestation, with fewer reports addressing the impact of paternal exposure on the progeny. One previous report has dealt with the impact of EtOH exposure on peripubertal male rats as a model of teenage drinking and the deleterious effects on the offspring. We report here findings examining the effect of 2 mo of EtOH feeding on male animals as they progressed through puberty on their ability to impregnate EtOH-naive female rats and characteristics of the subsequent litters. The EtOH-imbibing fathers weighed significantly less than pairfed controls and animals ingesting a non-EtOH liquid diet ad libitum. Nevertheless, they were able to mate successfully, although fecundity was significantly reduced. The number of successful pregnancies, defined as carried to term, was diminished from 92% in controls to 75% in EtOH-fed animals (p < 0.05). There was increased paternal testicular oxidative injury demonstrated by enhanced lipid peroxidation, protein oxidation, and decreased ratio of reduced to oxidized glutathione. The litter size of the EtOH-exposed males was reduced by 46%. The average litter size was 12.4+/-1.5 pups/litter in ad libitum animals, virtually identical to the 12.5+/-0.6 pups/litter in the pair fed controls. This is in sharp contrast to the 6.7+/-0.1 pups/litter from the paternal EtOH matings (p < 0.001). There was an increase in the average individual weight of pup offspring of paternally EtOH-exposed animals (p < 0.01 vs pair-fed controls and p < 0.05 vs ad libitum). Curiously, the male-to-female pup ratio was altered with a higher preponderance of male offspring from EtOH-fed fathers. There were no gross malformations noted among the pups. Insulin-like growth factor-1 levels in the pups at 10 d of age were unaltered between the groups. However, leptin was significantly elevated in the EtOH offspring. It appears that chronic EtOH exposure in the peripubertal fathers subsequently decreases fecundity and that this may be mediated by testicular oxidative injury, perhaps leading to accelerated germ cell apoptosis. PMID- 11394641 TI - Hyperleptinemia in pregnant bats is characterized by increased placental leptin secretion in vitro. AB - Hyperleptinemia is a common feature of pregnancy in mammals. The source of increased plasma leptin is uncertain. We examined leptin secretory rates in vitro to test the hypothesis that leptin secretion is upregulated during pregnancy. Two species of insectivorous bats were examined, Myotis lucifugus and Eptesicus fuscus, because of their unique reproductive cycle. Body mass and plasma leptin significantly increased with gestation and decreased during lactation. Adiposity increased in midgestation, then decreased in late gestation and lactation and was not significantly correlated with plasma leptin in pregnant or early lactating individuals. Leptin secretion in vitro per gram of adipose tissue tended to increase with gestation but was not significantly correlated with plasma leptin in the same individuals. Leptin secretion from placentae, however, increased with gestation and was significantly correlated with plasma leptin from the same individuals. In suckling pups, plasma leptin was high shortly after birth, then decreased to low levels that were not correlated with adiposity thereafter. We conclude that in bats, the placenta is a major source of circulating leptin during pregnancy, and that adiposity and plasma leptin levels are decoupled during three different periods of intense metabolic demand (pregnancy, early lactation, and neonatal growth). PMID- 11394643 TI - 4-Hydroxytamoxifen differentially exerts estrogenic and antiestrogenic effects on discrete subpopulations of human breast cancer cells. AB - Functional heterogeneity within populations of breast cancer cells contribute to the seemingly paradoxical effects of antiestrogens and the development of antiestrogen "resistance." Our objectives were to determine the degree to which T 47D cells may respond inappropriately (positively) to the antiestrogen 4 hydroxytamoxifen (HOT) alone, and whether all cells that respond to the stimulatory effects of estradiol-17beta (E2) are inhibited by the addition of HOT. Single, living T-47D cells were transfected by microinjection with an estrogen response element (ERE)-driven luciferase reporter plasmid. Transfected cells were then treated with medium alone, HOT, E2 or a combination thereof on consecutive days, exposed to the substrate luciferin and subjected to quantification of photonic emissions reflective of ERE-stimulated activity. This analysis revealed a subpopulation of cells that exhibited increased ERE-driven photonic activity in response to HOT. In companion studies, E2-stimulated ERE activity was reversed (on average) with HOT addition. However, analysis of individual cells revealed that although HOT reduced photonic activity in the majority (89.2%) of E2-responsive cells, there was a small subset (10.8% of the population) that was stimulated by E2 + HOT cotreatment. Our data support the hypothesis that these cells possess an intrinsic "resistance" to antiestrogenic agents, and that this could contribute to the remodeling of tumor cell populations toward a "resistant" phenotype. PMID- 11394644 TI - Characterization of a steroidogenic factor-1-binding site found in promoter of sterol carrier protein-2 gene. AB - Sterol carrier protein-2 (SCP2) is thought to mediate intracellular cholesterol transport in steroidogenic tissues. To elucidate the mechanism underlying the expression of this gene, a 300-bp fragment of the SCP2 promoter was cloned and analyzed for regulatory motifs. This promoter region contained a SF-1 binding motif, three activator protein-1 elements, an insulin response element, and a peroxisomal proliferator response element. The putative SF-1 binding region reacted with recombinant SF-1 DNA-binding domain in a mobility shift assay. The SCP2 promoter fragment was linked to a luciferase reporter gene and cotransfected in the presence or absence of SF-1 into HTB-9 cells. The results indicated that SF-1 was able to increase SCP2 promoter activity, an effect that was enhanced by cAMP. Similar results were obtained when the SCP2 promoter construct was cotransfected into Y1 cells. Cotransfection studies carried out in Kin 8 cells, a Y1 cell line with a mutation that prevents cAMP activation of PKA, revealed that a functional PKA is required for cAMP induction of SCP2 gene transcription. These results demonstrated that SF-1 confers cAMP responsiveness to the SCP2 promoter suggesting that SF-1 activation may be critical in regulation of this cholesterol transport protein. PMID- 11394642 TI - Environmental toxicant effects on neuroendocrine function. AB - Exposure to environmental toxicants can have profound effects on normal growth and development. However, the mechanisms by which these toxicants exert these effects are not well understood. Many environmental toxicants alter reproductive function and have effects on the central nervous system and behavior, yet the link between these reproductive and neurologic phenomena has not been systematically investigated. The neuroendocrine (hypothalamic-pituitary-gonadal) axis, which integrates inputs to and outputs from the nervous and reproductive systems, is functionally and anatomically situated to mediate effects of environmental toxicants, particularly those that are endocrine-disrupting chemicals (EDCs), on developmental processes. This article reviews the current literature on EDC effects on the neuroendocrine system, particularly at the level of hypothalamic gonadotropin-releasing hormone (GnRH) neurons, the key cells involved in the regulation of reproductive function. The focus of this article is on two polychlorinated biphenyl mixtures (Aroclor 1221, Aroclor 1254) and two organochlorine pesticides (methoxychlor and chlorpyrifos). Some experimental data are presented for each of the four urban environmental toxicants on GnRH cells in vitro and in vivo. The results of in vitro experiments indicate that all four of the toxicants profoundly affect hypothalamic GnRH gene expression, cell survival, and neurite outgrowth, demonstrating direct effects of EDCs on a GnRH cell line. In in vivo experiments, three of the toxicants (Aroclor 1221, methoxychlor, and chlorpyrifos) caused significant alterations in GnRH mRNA levels in female rats. Both the in vitro and in vivo findings support the novel concept of chlorpyrifos as an EDC. The results, taken together with the literature, support the hypothesis that the neuroendocrine axis, and specifically GnRH neurons, are sensitive to urban environmental toxicants, and that reproductive and neurologic effects of EDCs may be mediated at this level of the hypothalamic-pituitary gonadal axis. PMID- 11394645 TI - Endothelin-like immunoreactivity in lactotrophs, gonadotrophs, and somatotrophs of rat anterior pituitary gland are affected differentially by ovarian steroid hormones. AB - It has been previously found that all hormone-producing phenotypes of the anterior lobe of the pituitary gland are capable of producing endothelin (ET) like substances. The aim of this study was to determine whether the expression of ET-1-like peptides in lactotrophs, gonadotrophs, and somatotrophs is influenced by different in vivo ovarian hormonal conditions. Anterior lobes of the pituitary gland were harvested from ovariectomized and ovarian steroid-replaced adult female rats 10-12 d after surgery. Quantitative immunocytochemistry was performed on enzymatically dispersed pituitary cells. The presence of ET-1-like immunoreactivity in prolactin-, luteinizing hormone-, or growth hormone-producing cells was demonstrated by double-label immunocytochemistry. The incidence of ET-1 immunopositive pituitary cells was unaffected by progesterone treatment alone. Estradiol replacement caused a modest decrease in the number of lactotrophs and somatotrophs expressing ET-1 but increased the incidence of ET-1 immunopositive cells among gonadotrophs. Combined treatment with estradiol and progesterone robustly increased the incidence of ET-1 immunopositive lactotrophs and gonadotrophs but had no effect on somatotrophs. These data reveal that the synthesis of ET-1-like peptides in lactotrophs and gonadotrophs (and, to a lesser extent, in somatotrophs) is sensitive to ovarian steroids. Furthermore, these findings predict that ovarian steroids modulate ET-1 biosynthesis during the estrous cycle, suggesting a possible mechanism by which the ovarian steroid milieu may regulate the responsiveness of lactotrophs and gonadotrophs to their hypothalamic secretagogues. PMID- 11394646 TI - Neuropeptide Y in central control of feeding and interactions with orexin and leptin. AB - Neuropeptide (NPY) increases feeding when injected into the brain. In this study, we tested the hypothesis that its action might be related to feeding regulation of the orexin and leptin systems in rats. Intracerebroventricular administration of NPY (1 nmol/5 microL) stimulated feeding in rats. Injection of an antibody to orexin-A inhibited feeding, suggesting that endogenous orexin exerts a stimulatory tone on feeding. Intracerebroventricular injection of orexin antiserum before injection of NPY significantly attenuated the feeding response to NPY. On the other hand, ip pretreatment with leptin (2 mg/kg) significantly decreased food intake and inhibited NPY-induced feeding. We then examined whether orexin-containing neurons are activated under the stimulation of feeding in response to intracerebroventricular NPY or suppression of feeding in response to ip leptin, using Fos-like immunoreactivity (FLI) as a marker of neural activation. We observed that FLI was induced in the paraventricular, supraoptic, and dorsomedial nuclei as well as the lateral hypothalamic area (LHA) following administration of NPY. Double staining with anti-Fos and antiorexin antibodies revealed that 23.4% of the orexin-containing neurons in the LHA expressed FLI after NPY injection. Approximately 7.8% of the orexin-positive neurons in the LHA coexpressed Fos after leptin plus NPY. Our data indicate that a functional interaction among NPY, orexin, and leptin exists that may contribute to the central regulation of appetite. PMID- 11394647 TI - Hydrophilic bile salts enhance differential distribution of sphingomyelin and phosphatidylcholine between micellar and vesicular phases: potential implications for their effects in vivo. AB - BACKGROUND/AIMS: The hepatocyte canalicular membrane outer leaflet contains both phosphatidylcholine (PC) and sphingomyelin (SM). Normally, PC is the exclusive phospholipid in bile. We examined effects of bile salt hydrophobicity on cytotoxicity and on differential SM and PC distribution between detergent resistant aggregated vesicles (model for detergent-resistant canalicular membrane) and mixed micelles or small unilamellar vesicles (representing lipid phases in bile). METHODS: Aggregated vesicles were obtained by ultracentrifugation of cholesterol-supersaturated model systems containing SM, PC and various bile salts, micelles by ultrafiltration and unilamellar vesicles by dialysis of the supernatant. Erythrocyte hemolysis and lactate dehydrogenase release from CaCo-2 cells upon incubation with various micelles were quantified. RESULTS: Preferential SM distribution and lipid solubilization in aggregated vesicles increased in rank order taurodeoxycholate < taurocholate < tauroursodeoxycholate < taurohyodeoxycholate, with reciprocal PC enrichment in micelles and small unilamellar vesicles. Including small amounts of PC within taurohyodeoxycholate micelles increased cytotoxicity with more erythrocyte hemolysis and LDH release from CaCo-2 cells upon incubation, but decreased cytotoxicity in case of tauroursodeoxycholate micelles. CONCLUSIONS: Hydrophilic but not hydrophobic bile salts preserve integrity of pathophysiologically relevant phosphatidylcholine plus sphingomyelin-containing bilayers. Enhanced biliary phospholipid secretion during taurohyodeoxycholate but not during tauroursodeoxycholate therapy (Hepatology 25 (1997) 1306) may relate to different interactions of these bile salts with phospholipids. PMID- 11394648 TI - Increased expression of WAF1 in intrahepatic bile ducts in primary biliary cirrhosis relates to apoptosis. AB - BACKGROUND/AIMS: In primary biliary cirrhosis (PBC), the intrahepatic small bile ducts are selectively damaged by immune attacks, followed by progressive loss mainly due to apoptosis. Compared to the intercellular signaling such as the CD95/CD95 ligand interaction, little is known about alterations in intracellular cell cycle regulatory proteins and genotoxic damage in this apoptotic process. WAF1 is a potent and reversible inhibitor of cell cycle progression at both the G1 and G2 checkpoint and upregulated WAF1 induces irreversible G1 arrest and apoptosis. Transcriptional activation of the WAF1 gene is induced by the upregulated p53 in response to DNA damage. In this study, the cell cycle regulatory process of apoptosis in PBC was examined with respect to expression of WAF1. METHODS: Immunostaining for WAF1 and p53 was performed using 11 liver sections of PBC and 26 control livers. In addition, Ki67, apoptosis (TUNEL positive), and human telomerase RNA (hTR) were also detected. RESULTS: WAF1 was expressed in the nuclei of several epithelial cells in most damaged bile ducts in PBC but infrequently or rarely in controls. Some of these cells were also positive for p53, while the remainder were not. Ki67 immunostaining and TUNEL disclosed that the bile ducts in PBC showed increased cell division as well as enhanced apoptosis. Immunostaining of Ki67 and TUNEL staining showed that WAF1 positive cells were not proliferating, while some WAF1-positive cells were undergoing apoptosis. Moreover, the bile ducts lacked hTR expression, implying progressive shortening of telomeres during increased cell divisions. CONCLUSIONS: It seems possible that in PBC, expression of WAF1 on biliary epithelial cells relates to the apoptosis. p53 may be involved in this upregulation. This may be due to physiological upregulation of WAF1 and p53 in response to genotoxic damage such as oxidative stress associated with cholangitis, suggesting other processes than CD95/CD95 ligand interaction in biliary epithelial apoptosis in PBC. PMID- 11394649 TI - Acidosis modifies metabolic functions but does not affect vascular resistances in perfused rat livers. AB - BACKGROUND: Few data exist concerning the consequences of acidosis on intrahepatic vascular resistances and hepatic functions. METHODS: The consequences of pH and PCO2 changes on the intrahepatic vascular reactivity to norepinephrine (NE, 10(-9) to 3 x 10(-5) M) have been investigated in isolated rat livers perfused with solutions bubbled with 5, 10, or 15% CO2 and in solutions in which pH was decreased by replacing HCO3- with NaCl while maintaining a normal PCO2. Hepatic O2 consumption (VO2) and urea release were also measured during these experiments. RESULTS: The NE-induced increase of portal pressure did not change during hypercarbic and normocarbic acidosis. In contrast, the NE-induced increase of urea release was higher when the solution of perfusion was bubbled with 10 and 15% CO2, while during normocarbic acidosis the NE-induced increase of urea release did not change with pH. In the absence of NE, acidosis decreased hepatic VO2 and urea release but portal pressure was not modified by changing % CO2 or pH in the Krebs-Henseleit-bicarbonate solution. CONCLUSIONS: This study clearly shows that, in the liver, the consequences of acidosis are far more important on the metabolism (VO2 and urea release) than on the intrahepatic vascular resistance. PMID- 11394650 TI - Expression and cellular localization of fibrillin-1 in normal and pathological human liver. AB - BACKGROUND: The expression and the distribution of fibrillin-1 and elastin were studied in normal and pathological human liver samples. METHODS: As controls, histologically normal/subnormal liver samples (n = 24) were used. Pathological samples corresponded to seven cirrhosis and eight hepatocellular carcinomas (HCC) developed on cirrhotic (four) or noncirrhotic (four) liver. RESULTS: In normal liver, fibrillin-1 and elastin co-localized in vessel walls and portal tract connective tissue. Fibrillin-1 alone was detected along sinusoids and in portal spaces at the interface with the limiting hepatocytic plates and close to the basement membrane of bile ducts. By transmission electron microscopy, typical bundles of microfibrils were detected both in Disse space and in portal zones. Cirrhotic nodules were usually rich in fibrillin-1 along sinusoids; fibrillin-1 and elastin were co-localized in fibrotic septa surrounding nodules. In HCC, fibrillin-1 was present between tumoral hepatocytes; stromal reaction around the tumors contained both fibrillin-1 and elastin. CONCLUSIONS: Fibrillin-1 was associated with elastin in portal mesenchyme and vessel walls of normal liver, in fibrotic septa around cirrhotic nodules and stromal reaction around HCC, but was expressed alone in the perisinusoidal space. The functional roles for fibrillin-1 in non-elastic tissues, such as the liver, remain to be elucidated. PMID- 11394651 TI - Frequency and biochemical expression of C282Y/H63D hemochromatosis (HFE) gene mutations in the healthy adult population in Italy. AB - BACKGROUND/AIMS: The actual prevalence of the main hemochromatosis (HFE) mutations in the Italian adult population and their phenotypic expression have not yet been established. This information is key to advocate a mass-screening program. METHODS: Two thousand one hundred adults were tested for the C282Y/H63D HFE gene mutations by an automated genotyping assay as well as transferrin saturation (TS) and serum ferritin levels. RESULTS: No homozygotes for the C282Y mutation were found. Heterozygosity for the C282Y mutation was 3.1%, while heterozygosity and homozygosity for the H63D mutation were 21.5% and 2.5%, respectively. TS was significantly higher in C282Y heterozygotes and H63D homozygotes as compared to wild-type individuals (P < 0.01). Interestingly, of the HFE wild-type subjects 5.9% had a TS value above the 45% threshold. CONCLUSIONS: This study shows that (i) the predicted prevalence for C282Y homozygosity in Italy is 1:3900; (ii) the C282Y/H63D wild-type population has an increased baseline of iron parameters possibly due to genetic factors not linked to the C282Y/H63D mutations; (iii) since in the latter population the actual tissue iron burden cannot be assessed, phenotypic (TS) screening in Italy is not recommended until the true prevalence of all mutations in the HFE gene and in other hemochromatosis genes will be established. PMID- 11394652 TI - A selective ROCK inhibitor, Y27632, prevents dimethylnitrosamine-induced hepatic fibrosis in rats. AB - BACKGROUND: p160ROCK is a direct Rho target which mediates Rho-induced assembly of focal adhesions and stress fibers. We previously reported that Rho signaling pathways are involved in the activation of hepatic stellate cells (HSC) in vitro. The aim of the present study was to test the hypothesis that an inhibitor specific for p160ROCK (Y27632) could prevent experimental hepatic fibrosis induced by dimethylnitrosamine (DMN) in rats. METHODS: Y27632 was given orally at 30 mg/kg daily for 4 weeks after the first injection of DMN. The degree of fibrosis was evaluated by image analysis and also by measurements of collagen and hydroxyproline content in the liver. The expression of alpha-smooth muscle actin (alpha-SMA) in the liver and in the primary cultured HSC was also evaluated. Semi quantitative RT-PCR was performed to evaluate the expression of type I collagen mRNA in the liver. RESULTS: Y27632 treatment significantly decreased the occurrence of DMN-induced hepatic fibrosis and reduced the collagen and hydroxyproline content and alpha-SMA expression in the liver. The expression of alpha-SMA in HSC was also suppressed in vitro. CONCLUSIONS: These findings indicate that inhibitors of the Rho-ROCK pathway might be useful therapeutically in hepatic fibrosis. PMID- 11394653 TI - Eotaxin expression and eosinophil infiltrate in the liver of patients with drug induced liver disease. AB - BACKGROUND: Drug-induced liver disease is due to intrinsic or idiosyncratic hepatotoxins. Liver parenchyma is then infiltrated by immunocompetent cells. Eosinophils are primarily tissue leukocytes which are attracted into tissues by various chemoattractants, including chemokines. The aim of this study was to study eosinophils in the livers of patients with drug-induced liver disease. METHODS: Immunohistochemical studies with antibodies against eosinophil cationic proteins (major basic protein, eosinophil-derived neurotoxin and eosinophil cationic protein), cytokines (interleukin 5 (IL-5), interleukin 3 (IL-3) and granulocyte-macrophage colony stimulating factor (GM-CSF)) and chemokines (eotaxin and RANTES (Regulated upon Activation Normal T cell Expressed and Secreted)) were performed to assess the in situ activation of the liver infiltrating eosinophils of 14 patients with drug-induced liver disease and 19 controls. RESULTS: Eosinophils were only observed in patients with drug-induced liver disease. Eosinophils were morphologically normal when hepatitis was due to paracetamol whereas eosinophils had granular changes when hepatitis was due to an idiosyncratic hepatotoxin. Eotaxin was detected in all patients with drug-induced liver disease, whereas RANTES was detected in three of them. IL-5, IL-3 or GM-CSF were not detected. CONCLUSIONS: In patients with drug-induced liver disease, the recruitment of eosinophils in the liver may depend on eotaxin expression. Eosinophil changes may vary according to the type of drug. PMID- 11394654 TI - Cerebral hyperemia and nitric oxide synthase in rats with ammonia-induced brain edema. AB - BACKGROUND/AIM: Brain edema is a common fatal complication in acute liver failure. It is related to an acute change in brain osmolarity secondary to the glial accumulation of glutamine. Since high cerebral blood flow (CBF) precedes cerebral herniation in fulminant hepatic failure we first determined if an increase in brain water and glutamine are prerequisite to a rise in CBF in a model of ammonia-induced brain edema. Secondly, we determined if such a cerebral hyperperfusion is mediated by nitric oxide synthase (NOS). METHODS: Male rats received an end-to-side portacaval anastomosis (PCA). At 24 h, they were anesthetized with ketamine and infused with ammonium acetate (55 microM/kg per min). Studies were performed at 60, 90, 120, 150 and 180 min after starting the ammonia infusion and once the intracranial pressure had risen three-fold (mean 210'). Brain water (BW) was measured using the gravimetry method and CBF with the radioactive microsphere technique. Glutamine (GLN) in the CSF was sampled via a cisterna magna catheter. The neuronal NOS was specifically inhibited by 1-2 trifluoromethylphenyl imidazole (TRIM, 50 mg/kg intraperitoneally) and in separate studies nonspecifically by N-omega-nitro-L-arginine (L-NNA, 2 microg/kg per min intravenously) RESULTS: At 90', brain water was significantly increased (P < 0.015) as compared to the 60' group while CBF was significantly different at 150'. A significant correlation was observed between values of CBF and brain water (r = 0.88, n = 36, P < 0.001). Administration of either TRIM or L-NNA did not prevent the development of cerebral hyperperfu. sion and edema. CONCLUSION: We observed that cerebral hyperemia follows an initial rise in brain water content, rather than in the cerebrospinal fluid concentration of glutamine. The rise in CBF further correlated with brain water accumulation and was of critical importance for the development of intracranial hypertension. The unique mechanism for the rise in CBF in hyperammonemia was not prevented by NOS inhibition indicating that NO is not the mediator of high CBF and intracranial hypertension. PMID- 11394655 TI - Differential expression of CD44 isoforms during liver regeneration in rats. AB - BACKGROUND: CD44 is a transmembrane glycoprotein known to bind hyaluronic acid (HA). This molecule is a multifunctional cell surface glycoprotein involved in lymphocyte homing and activation, tumor growth and metastasis. We have investigated the qualitative modification of CD44 in the regenerating liver as a model for studying cellular proliferation in vivo. Molecules involved in cell adhesion and the extracellular matrix (ECM), which influence differentiation, growth, cell-cell interactions and cellular polarity, play an important role in the liver regeneration. We studied the modulation of CD44 gene expression and its post-transcriptional modifications, analyzing the expression of different isoforms containing exon v6 in the regenerating liver, in sham operated liver and in the hepatoma cells H-35. METHODS: The expression of CD44 and CD44v6 were analyzed in RNA extracted from regenerating liver at different times after partial hepatectomy (PH), and H-35 hepatoma cells by Northern blot, RT-PCR and Southern blot, and in protein extracts from regenerating liver by Western blot. H 35 hepatoma cells were assayed with the antibody cross-linked technique with CD44 antibodies. RESULTS: The standard CD44 form is expressed in regenerating liver and its levels were not modified following PH. However, our analysis revealed CD44 isoforms containing v6 in the first hours after PH as well as in the H-35 hepatoma cell line. H-35 cells treated with cross-linked anti-CD44 antibodies or HA show an increased rate of incorporation of [3H]thymidine (30 and 25%, respectively) with respect to the control. CONCLUSION: These findings suggest that CD44 may play a role in the proliferation of residual hepatocytes following PH. PMID- 11394656 TI - Preliminary results of interferon-alpha therapy on woodchuck hepatitis virus induced hepatocarcinogenesis: possible benefit in female transgenic mice. AB - BACKGROUND: C-myc activation is a potent oncogenic event in hepatocarcinogenesis. The aim of this study was to test the preventive effect of interferon-alpha (IFN alpha) on the development of dysplasia and subsequent hepatocellular carcinoma (HCC) in transgenic (Tg) mice overexpressing c-myc in the liver. METHODS: The WHV/c-myc Tg mice recapitulating woodchuck hepatitis virus-induced hepatocarcinogenesis were treated with IFN-alpha, starting early in life until sacrifice at pre-neoplastic or neoplastic stages. Transgene expression was assessed by reverse transcription-polymerase chain reaction (RT-PCR), hepatocyte proliferation was assessed by bromodeoxyuridine incorporation and RT-PCR for proliferating cell nuclear antigen, and apoptosis was assessed by in situ nick end-labeling of DNA. RESULTS: C-myc expression and hepatocyte proliferation were significantly reduced in treated female mice, without modification of apoptosis, correlating with a lower severity of dysplasia in 9/12 treated animals at pre neoplastic stages. At the neoplastic stage, 2/3 treated females neither exhibited carcinoma nor dysplasia, while all 6/6 untreated mice and 3/3 treated males developed carcinomas. CONCLUSIONS: Inhibition of c-myc and hepatocyte proliferation by long-term administration of IFN-alpha was associated with a decrease, or a delay, of oncogenesis in the mouse Tg HCC model. Whether c-myc and hepatocyte proliferation down-regulation could be relevant parameters of IFN alpha efficiency for hepatocarcinogenesis prevention in cirrhotic patients should be established. PMID- 11394657 TI - Serum alpha-fetoprotein for diagnosis of hepatocellular carcinoma in patients with chronic liver disease: influence of HBsAg and anti-HCV status. AB - BACKGROUND: It is not established whether virological status affects the efficiency of alpha-fetoprotein (AFP) as a hepatocellular carcinoma (HCC) marker among patients with chronic liver disease (CLD). METHODS: We enrolled in a case control study 170 HCC and 170 CLD patients, matched for age, sex, CLD and HBsAg/anti-HCV status. The AFP sensitivity, specificity, positive (PPV) and negative (NPV) predictive values were calculated. PPV and NPV were evaluated for three additional HCC prevalences (5, 10, and 20%). RESULTS: The best discriminating AFP value was 16 ng/ml. A value of 20 ng/ml (above which investigations for HCC are recommended) had equivalent sensitivity (60.0 vs. 62.4%) and specificity (90.6 vs. 89.4%). PPV of 20 ng/ml was 84.6% but decreased to 25.1% at 5% tumor prevalence. NPV was 69.4% and rose to 97.7% at 5% prevalence. In the different groups of infected patients PPV ranged from 80.0 to 90.9%, falling to 17.4-34.5% at 5% prevalence. In noninfected patients PPV was 100% at any HCC prevalence. NPV ranged from 59.0 to 73.0%, reaching 96.5-98.1% at 5% prevalence. CONCLUSIONS: In CLD patients, AFP monitoring misses many HCCs and inappropriately arouses suspicion of malignancy in many patients. Its usefulness is barely affected by the infection responsible for CLD. An AFP elevation could be more indicative of HCC in non-infected patients. PMID- 11394658 TI - Characterization of serum activin-A and follistatin and their relation to virological and histological determinants in chronic viral hepatitis. AB - BACKGROUND/METHODS: Hepatocyte proliferation in viral hepatitis is regulated by a number of growth factors. Activin-A inhibits hepatocyte DNA synthesis while follistatin, a potent activin-A antagonist, promotes liver regeneration. We report the first study of activin-A and follistatin in human viral hepatitis. Sera from 15 normal subjects, 22 hepatitis B and 47 hepatitis C patients were analysed for activin-A and follistatin and correlated with serological and histological markers of liver injury and with specific immunohistochemistry. RESULTS: All groups showed immunoreactivity for activin with hepatocyte localisation. Serum activin-A was significantly increased in viral hepatitis patients compared to controls, was greater in hepatitis B compared to hepatitis C, and correlated with serum aminotransferase and hepatitis B viral replication. A concurrent rise in serum follistatin was not observed in either group, but serum follistatin correlated inversely with hepatitis B DNA levels. Although hepatocyte apoptosis in hepatitis C and proliferation in both groups was significantly elevated compared to controls, there was no correlation with serum activin-A or follistatin. CONCLUSIONS: Activin-A and follistatin are constitutively expressed in human liver and serum concentrations are increased in viral hepatitis. Dysregulation of the activin/follistatin axis may be linked to hepatitis B replication but does not correlate with hepatocyte apoptosis. PMID- 11394659 TI - Detection of YMDD motif mutations in some lamivudine-untreated asymptomatic hepatitis B virus carriers. AB - BACKGROUND/AIM: It is well documented that long-term lamivudine treatment induces emergence of lamivudine-resistant hepatitis B virus (HBV), namely, YMDD motif mutation in some patients chronically infected with HBV. We previously reported that there were no YMDD mutant viruses in patients with chronic hepatitis B who were not treated with lamivudine. In this series, we examined mutations in the YMDD motif gene in asymptomatic carriers who maintained normal ALT values for 1 year or more. METHODS: Serum samples obtained from 18 patients chronically infected with HBV who consulted our university were used. None of these patients had any experience of using antiviral agents. For detection of mutant viruses, a kit developed in our laboratory was used. RESULTS: Mutations were detected in five of 18 samples: YMDD+YIDD in three samples and YMDD+YVDD+YIDD in two samples. All of these five samples were positive for anit-HBe. In five samples in which mutations were observed, sequencing was carried out following subcloning. CONCLUSION: The present study demonstrates that YMDD mutant viruses are present in lamivudine-untreated asymptomatic hepatitis B virus carriers as well. PMID- 11394660 TI - DNA oxidative damage in leukocytes correlates with the severity of HCV-related liver disease: validation in an open population study. AB - BACKGROUND/AIMS: Oxidative DNA damage, identifiable in the formation of 8 hydroxydeoxyguanosine (8-OHdG), is relevant in the mutagenesis/carcinogenesis process. The aim of this study was to assess 8-OHdG levels in patients with hepatitis C virus (HCV) infection in relation to extent of liver damage and HCV genotype. METHODS: 8-OHdG levels were measured in DNA from circulating leukocytes of 110 anti-HCV positive subjects belonging to the population of the Dionysos study, subgrouped in: 50 anti-HCV+ with persistently normal ALT, 48 with chronic hepatitis and 12 with cirrhosis. Twenty normal subjects served as Controls. 8 OHdG levels were assayed by HPLC/electrochemical detector. RESULTS: 8-OHdG levels rose (P < 0.00001) from Controls to HCV+; chronic hepatitis and cirrhosis were associated with a further increase (P < 0.02 versus HCV+). Genotype 1 was associated with higher levels of 8-OHdG (P < 0.04). Multiple logistic regression analysis showed that, after correction for potential confoundings, 8-OHdG levels correlated (P < 0.02) with presence and extent of liver damage. CONCLUSIONS: An accumulation of 8-OHdG in circulating leukocytes is a reliable marker of the extent of liver damage in HCV+ patients and is present in particular in genotype 1 infection. This genomic damage may contribute to liver carcinogenesis by causing persistent DNA changes. PMID- 11394661 TI - Interferon and prevention of hepatocellular carcinoma in viral cirrhosis: an evidence-based approach. AB - BACKGROUND/AIMS: To evaluate by meta-analysis of available literature whether interferon (IFN) reduces the incidence of hepatocellular carcinoma (HCC) in patients with hepatitis B virus (HBV) or hepatitis C virus (HCV)-related Child A cirrhosis. METHODS: Three randomized controlled trials and 15 nonrandomized controlled trials, including 4614 patients and comparing IFN to no treatment, were selected. Data on the incidence of HCC in IFN treated and untreated patients were extracted from each study. Meta-analysis by the DerSimonian and Laird risk difference (RD) method was used to pool observations. RESULTS: A different incidence of HCC between treated and untreated cirrhotic patients was observed for HCV (overall RD -12.8%; 95% CI -8.3 to -17.2%, P < 0.0001) and HBV (overall RD -6.4%; 95% CI -2.8 to -10%, P < 0.001). In HCV-related cirrhosis, the rate of HCC development was lower in sustained responders to IFN than in untreated patients (overall RD -19.1%; 95% CI -13.1 to -25.2%, P < 0.00001), with low heterogeneity among trials (P=0.053), and also in nonresponders vs. untreated patients (overall RD -11.8%; 95% CI -6.4 to -19.1%, P < 0.0001), although with significant heterogeneity. Inconsistency among the studies was a major problem, both for HCV (chi2 = 58.16 with 13 DF; P < 0.0001) and HBV (chi2 = 26.4 with 6 DF; P = 0.0001) related cirrhosis, and also when follow-up was shorter than 60 months. Consistent results were only observed when assessing data from European reports: in this subgroup no preventive effect of HCC was shown for HBV (overall RD -4.8%; 95% CI -11.1-1.5%, P, not significant), and only a weak effect for HCV (overall RD -10%; 95% CI -5.9 to -14.2%; P < 0.0001). CONCLUSIONS: Literature data pooling suggests a slight preventive effect of IFN on HCC development in patients with HCV-related cirrhosis. The magnitude of this effect is low and the observed benefit might be due to spurious associations. The preventive effect is more evident among sustained responders to IFN. IFN does not seem to affect the rate of HCC in HBV-related cirrhosis. PMID- 11394662 TI - Alphafetoprotein: an obituary. PMID- 11394663 TI - Chemoprevention of hepatocellular carcinoma in hepatitis C virus-related cirrhosis: first, eliminate the virus. PMID- 11394664 TI - Hepatic encephalopathy and nitric oxide. PMID- 11394665 TI - Live donor liver transplantation. PMID- 11394666 TI - Allogenic bone marrow transplantation with a donor presenting with an acute hepatitis A. AB - We report the unique occurrence of an allogenic bone marrow transplantation performed as the donor was suffering from an acute hepatitis A. The bone marrow was contaminated at the time of collection, as demonstrated by hepatitis A virus (HAV) RNA detected by RT-PCR. Hepatitis A virus infection in such a situation could have resulted in a severe liver disease in the recipient. However, although we could demonstrate that the recipient had been infected, he did not develop a symptomatic hepatitis A but only minor disturbances of liver function tests between days 35 and 55. Both the postponement of the transplantation and the use of intravenous polyvalent immunoglobulins have probably played a key role in decreasing the viral load and allowing a rapid clearance of the virus. A possible role of the grafted immune system might also be envisaged, as suggested by the de novo synthesis of IgM in the recipient. PMID- 11394667 TI - Primary sclerosing cholangitis associated with rheumatoid arthritis and HLA DR4: is the association a marker of patients with progressive liver disease? AB - In four cases we describe the unique association of primary sclerosing cholangitis (PSC) and rheumatoid arthritis (RA). In three of the cases the liver disease was unusually progressive, proceeding to cirrhosis in 14, 18 and 48 months from diagnosis. The three cases with progressive liver disease and ulcerative colitis were all HLA type DR4. The fourth patient also suffered from coeliac disease in addition to PSC and RA and has remained asymptomatic over 7 years of follow-up. RA in association with PSC may serve as a clinical marker of patients at high risk of progression to cirrhosis who need to be kept under particularly close observation. In addition, PSC needs to be considered in the differential diagnosis of all patients with RA and cholestatic liver function tests. This is especially important given the link between PSC and an increased risk of colonic carcinoma, and thus the need for surveillance colonoscopy. PMID- 11394668 TI - Variation in UGT1A1 activity in Gilbert's syndrome. PMID- 11394669 TI - Learning of event sequences is based on response-effect learning: further evidence from a serial reaction task. AB - Four experiments provide converging evidence that serial learning in a serial reaction task is based on response-effect learning, mediated by the learning of the relations between a response and the stimulus that follows it. In Experiment 1, the authors varied the stimulus sequence and the response-stimulus relations while holding the response sequence constant. Learning effects depended on the complexity of the response-stimulus relations but not on the stimulus-stimulus relations. In Experiment 2, transfer of serial learning from 1 stimulus sequence to another was only found when both sequences had identical response-stimulus relations. In Experiment 3, a variation of the stimulus sequence alone had no effect on serial learning, whereas in Experiment 4 learning effects increased when the response-stimulus relations but not the stimulus-stimulus relations were simplified. These findings suggest that serial learning is based on mechanisms of voluntary action control. PMID- 11394670 TI - Two modes of transfer in artificial grammar learning. AB - Participants can transfer grammatical knowledge acquired implicitly in 1 vocabulary to new sequences instantiated in both the same and a novel vocabulary. Two principal theories have been advanced to account for these effects. One suggests that sequential dependencies form the basis for cross-domain transfer (e.g., Z. Dienes, G. T. M. Altmann, & S. J. Gao, 1999). Another argues that a form of episodic memory known as abstract analogy is sufficient (e.g., L. R. Brooks & J. R. Vokey, 1991). Three experiments reveal the contributions of the 2. In Experiment 1 sequential dependencies form the only basis for transfer. Experiment 2 demonstrates that this process is impaired by a change in the distributional properties of the language. Experiment 3 demonstrates that abstract analogy of repetition structure is relatively immune to such a change. These findings inform theories of artificial grammar learning and the transfer of grammatical knowledge. PMID- 11394671 TI - On attentional control as a source of residual shift costs: evidence from two component task shifts. AB - It is widely assumed that supervisory or attentional control plays a role only in the preparatory reconfiguration of the mental system in task shifting. The well known fact that residual shift costs are still present even after extensive preparation is usually attributed to passive mechanisms such as cross talk. The authors question this view and suggest that attentional control is also responsible for residual shift costs. The authors hypothesize that, under shift conditions, tasks are executed in a controlled mode to guarantee reliable performance. Consequently, the control of 2 task components should require more resources than the control of only 1. A series of 4 experiments with 2-component tasks was conducted to test this hypothesis. As expected, more residual shift costs were observed when 2 components rather than 1 varied across trials. Interference effects and sequential effects could not account for these results. PMID- 11394672 TI - Habituation to auditory distractors in a cross-modal, color-word interference task. AB - The speed of naming the color of a colored square was examined with acoustic distraction to study the effects of the formation of a mental representation (neural model) of distractors. In Experiment 1, practice with the distractors (color words, noncolor words, and tones) was examined, and in Experiments 2 and 3 each color-naming trial was preceded by preexposures to sounds that could be dissimilar, similar, or identical to the upcoming auditory distractor. Consistency in the identity of ignored sounds, whether during color-naming practice or between preexposures and test, reduced interference with color naming. Consistency in voice played no role, and attended preexposures were ineffective in reducing interference. Given these results, the authors propose that mental representations of distractors include information about their task relevance, which modulates disruption of the primary task. PMID- 11394673 TI - Parallel memory retrieval in dual-task situations: II. Episodic memory. AB - Three experiments asked whether subjects could retrieve information from a 2nd stimulus while they retrieved information from a 1st stimulus. Subjects performed recognition judgments on each of 2 words that followed each other by 0, 250, and 1,000 ms (Experiment 1) or 0 and 300 ms (Experiments 2 and 3). In each experiment, reaction time to both stimuli was faster when the 2 stimuli were both targets (on the study list) or both lures (not on the study list) than when 1 was a target and the other was a lure. Each experiment found priming from the 2nd stimulus to the 1st when both stimuli were targets. Reaction time to the 1st stimulus was faster when the 2 targets came from the same memory structure at study (columns in Experiment 1; pairs in Experiment 2; sentences in Experiment 3) than when they came from different structures. This priming is inconsistent with discrete serial retrieval and consistent with parallel retrieval. PMID- 11394674 TI - Proactive interference, accessibility bias, and process dissociations: valid subjective reports of memory. AB - Proactive interference was assessed with a variant of the process-dissociation procedure, which separates effects of habit (accessibility bias) and recollection (discriminability). In three cued-recall experiments, proactive interference was shown to be an effect of bias rather than an effect on actual remembering. Divided attention, age, and study duration selectively influenced the recollection parameter, whereas training probability selectively influenced the habit parameter. Furthermore, in Experiments 2 and 3, subjective reports of remembering were highly correlated with, and nearly identical to, objective estimates of recollection gained from the process-dissociation procedure. The authors discuss the relevance of the results to theories of proactive interference and argue that older adults' greater susceptibility to interference effects is sometimes caused by an inability to recollect rather than by an inability to inhibit a preponderant response. PMID- 11394675 TI - On the nature of associative information in recognition memory. AB - In a typical associative-recognition task, participants must distinguish between intact word pairs (both words previously studied together) and rearranged word pairs (both words previously studied but as part of different pairs). The familiarity of the individual items on this task is uninformative because all of the items were seen before, so the only way to solve the task is to rely on associative information. Prior research suggests that associative information is recall-like in nature and may therefore be an all-or-none variable. The present research reports several experiments in which some pairs were strengthened during list presentation. The resulting hit rates and false alarm rates, and an analysis of the corresponding receiver operating characteristic plots, suggest that participants rely heavily on item information when making an associative recognition decision (to no avail) and that associative information may be best thought of as a some-or-none variable. PMID- 11394677 TI - Interference from related items in object identification. AB - Interference between related items in the identification of objects was examined using a postcue procedure. Pairs of objects were presented as differently colored line drawings followed by a color cue to indicate which object to name. Naming latencies were longer when both objects were from the same superordinate category than when they were unrelated. This interference effect was replicated when subjects were cued to report the color of a drawing rather than its name. Interference was greatly reduced when more distinctive attributes were used to distinguish members of a pair, both when the task required naming an object and when it required report of an object's attribute. These results challenge accounts of interference in the postcue paradigm that are based on competitive interactions in the activation of phonological representations by semantics and instead implicate object-attribute integration in memory as the source of interference. PMID- 11394676 TI - The role of familiarity in item recognition, associative recognition, and plurality recognition on self-paced and speeded tests. AB - Four experiments compare the effect of familiarity on item, associative, and plurality recognition on self-paced and speeded tests. The familiarity of test items was enhanced by presenting a prime that matched the subsequent test item. On item and plurality recognition tests, participants were more likely to respond "old" to primed than to unprimed test items. In associative recognition, priming increased the proportion of old responses on a speeded test, but not on a self paced test. This suggests that familiarity plays a larger role in item and plurality recognition than in associative recognition on self-paced tests. On speeded tests, priming has a similar effect on item, associative, and plurality recognition. Results suggest that item and associative recognition rely differentially on familiarity and recollection. They are also consistent with recent evidence suggesting that different processes underlie plurality and associative recognition. PMID- 11394678 TI - A rose by any other name is not the same: the role of orthographic knowledge in homophone confusion errors. AB - Homophone confusion errors were examined in a series of 6 experiments. Across a variety of tasks, readers consistently made more errors on homophone trials than on control trials. These effects were established in Experiment 1 using a semantic-decision task in which participants judged whether pairs of words were related or unrelated. For both related and unrelated trials, error rates were higher for homophones as compared with controls. Results such as these have previously been taken as evidence for the role of phonology in lexical access and reading. However, differences in orthographic knowledge (more specifically, knowledge of spelling-to-meaning correspondences) across participants and homophone items significantly predicted homophone errors across all tasks. In addition, spelling tasks and multiple-choice questionnaires revealed differences in orthographic knowledge across participants and homophone items. Although these results do not rule out a role for phonology in lexical access, they indicate that homophone confusion errors may also be due to factors other than phonology. PMID- 11394679 TI - Are individual or consecutive letters the unit affected by repetition blindness? AB - When 2 similar words (e.g., react reach) are briefly sequentially displayed, the 2nd word may be omitted from the report, a phenomenon known as repetition blindness (RB). Previous researchers have suggested that consecutive letters are the unit affected by RB. Six experiments provided new data on orthographic RB. Two letters at the beginning or end of words resulted in RB, as did alternating interior letters (tactile earthly) and 3 letters with different relative positions (arid bird). However, no RB was found with a single final letter (show view). Observed RB may reflect pattern completion because RB for pairs like throat theory was reduced when the nonrepeated letters (eory) were consistent with only a single word. The experiments point to a model of orthographic RB in which both individual letters and letter sequences of length 2 or more play a role. PMID- 11394680 TI - Prototypes in category learning: the effects of category size, category structure, and stimulus complexity. AB - Although research in categorization has sometimes been motivated by prototype theory, recent studies have favored exemplar theory. However, some of these studies focused on small, poorly differentiated categories composed of simple, 4 dimensional stimuli. Some analyzed the aggregate data of entire groups. Some compared powerful multiplicative exemplar models to less powerful additive prototype models. Here, comparable prototype and exemplar models were fit to individual-participant data in 4 experiments that sampled category sets varying in size, level of category structure, and stimulus complexity (dimensionality). The prototype model always fit the observed data better than the exemplar model did. Prototype-based processes seemed especially relevant when participants learned categories that were larger or contained more complex stimuli. PMID- 11394681 TI - Categorization processes in mental comparisons. AB - Five experiments explore categorization and category-based congruity effects in mental comparisons. The first 4 experiments concentrate on categorization of infinite-set small items. The experiments vary the additional items presented and whether those items appear once (Experiments 1-2) or repeatedly (Experiments 3 4). Additional items include other small items (Experiment 1), relatively large items (Experiments 2-4), and items involving nonsize dimensions (Experiment 4). The critical small items show a complete congruity effect only in Experiments 1 and 3. Results suggest that categorization of infinite-set items may be based on range information alone (Experiment 1) but that multiple categorizations based on multiple ranges (Experiment 2) may require attentional effort. Results implicate categorization as a central process in mental comparison, despite differences in ease of categorization across paradigm. PMID- 11394683 TI - Distributing working memory resources during problem solving. AB - This study examines how problem solvers distribute working memory demands over internal and external resources. Participants recorded notes while performing an arithmetic task. They recorded a majority of intermediate results and labeled many of those results (e.g., "C = 10"). When more effort was required to take notes, participants recorded fewer results. Participants with a consistent goal structure recorded fewer results and with practice labeled fewer recorded results than those with varied goal structures. When notes were displayed in a consistent spatial arrangement participants labeled fewer recorded results than when notes appeared in varied locations. These findings indicate that individuals use explicit and implicit strategies for indexing intermediate results. The data support the view that individuals flexibly distribute working memory over internal and external resources in response to situational cost-benefit considerations. PMID- 11394682 TI - Working memory and focal attention. AB - Measures of retrieval speed indicated that only a small subset of representations in working memory falls within the focus of attention. An n-back task, which required tracking an item 1, 2, or 3 back in a sequentially presented list, was used to examine the representation and retrieval of recent events and how control processes can be used to maintain an item in focal attention while concurrently processing new information. A speed-accuracy trade-off procedure was used to derive measures of the availability and the speed with which recent events can be accessed. Results converge with other time course studies in demonstrating that attention can be concurrently allocated only to a small number of memory representations, perhaps just 1 item. Measures of retrieval speed further demonstrate that order information is retrieved by a slow search process when an item is not maintained within focal attention. PMID- 11394684 TI - High-level reasoning and base-rate use: do we need cue-competition to explain the inverse base-rate effect? AB - Previous accounts of the inverse base-rate effect (D. L. Medin & S. M. Edelson, 1988) have revolved around the concept of cue-competition. In this article, the authors propose that high-level reasoning in the form of an eliminative inference mechanism may contribute to the effect. A quantitative implementation of this idea demonstrates that it has the power by itself to produce the pattern of base rate effects in the Medin and Edelson (1988) design. Four predictions are derived that contradict the predictions by attention to distinctive input (ADIT; J. K. Kruschke, 1996), up to date the most successful account of the inverse base-rate effect. Results from 3 experiments disconfirm the predictions by ADIT and demonstrate the importance of high-level reasoning in designs of the Medin and Edelson kind. Implications for the interpretation of the inverse base-rate effect and the attention-shifting mechanisms presumed by ADIT are discussed. PMID- 11394685 TI - Proactive interference and complexity. AB - C. L. Isaac and A. R. Mayes (1999a, 1999b) compared forgetting rates in amnesic patients and normal participants across a range of memory tasks. Although the results are complex, many of them appear to be replicable and there are several commendable features to the design and analysis. Nevertheless, the authors largely ignored 2 relevant literatures: the traditional literature on proactive inhibition/interference and the formal analyses of the complexity of the bindings (associations) required for memory tasks. It is shown how the empirical results and conceptual analyses in these literatures are needed to guide the choice of task, the design of experiments, and the interpretation of results for amnesic patients and normal participants. PMID- 11394686 TI - Processes underlying long-term repetition priming in digit data entry. AB - Two experiments examined long-term repetition priming in data entry. In each experiment, participants entered 4-digit numbers displayed as either words or numerals, and responded with digits (Experiment 1), or either digits or initial letters (Experiment 2). At test 1 week later, they entered old and new numbers, with the format changed for half of the old stimuli. Implicit memory was evidenced at test by faster entry of the old than the new numbers, regardless of whether the numbers were in the same or different format, suggesting that the abstract numerical meaning, not the surface form, contributes to repetition priming. Numbers presented as words in training had an advantage over numbers presented as numerals, regardless of response format, implying that type of processing also contributes to the effect and ruling out an explanation based on time spent processing numbers in word format. PMID- 11394687 TI - Effects of dividing attention on the memory-block effect. AB - S. M. Smith and D. R. Tindell (1997) reported that prior study of words that are orthographically similar to the solutions of test word fragments (e.g., studying ANALOGY and completing the fragment A_L _ _GY, whose solution is ALLERGY) reduced the fragment completion rate relative to a baseline condition in which unrelated words were studied. They called this effect the memory-block effect. In the present experiment, the authors replicated this effect using a larger set of materials than that used by S. M. Smith and D. R. Tindell. The authors also found that dividing attention at study eliminated the memory-block effect. This pattern mimicked the effect of dividing attention on recognition memory but differed from the effect on repetition priming effects. The authors suggest that the memory block effect is driven by a mechanism different from that responsible for producing repetition priming effects in an implicit fragment completion test. PMID- 11394688 TI - Antigenic comparison of a truncated form of VP2 of infectious pancreatic necrosis (IPN) virus expressed in four different cell types. AB - A truncated form of the structural protein VP2 (truncVP2) of infectious pancreatic necrosis (IPN) virus encompassing amino acids 147-307 was expressed in bacterial, yeast, piscine and mammalian cells. All four recombinant antigens were recognised by a VP2-specific monoclonal antibody by ELISA and immunoblot analysis. However, the minimum amount of r-truncVP2 needed for detection by these methods varies depending on the cell type used for expression. Furthermore, all four recombinant preparations, when used to immunise Atlantic salmon, were capable of inducing antibodies reactive with whole IPNV in ELISA. PMID- 11394689 TI - Cloning and characterisation of tandem-repeat type galectin in rainbow trout (Oncorhynchus mykiss). AB - Fish beta-galactoside binding lectin (galectin) cDNA was cloned from the cDNA library of rainbow trout (Oncorhynchus mykiss) head kidney. The clone contained a single open reading frame encoding 341 amino acids (aa) (38 kDa protein), including the initiator methionine. Significant sequence homology to mammalian galectin-9 (40-55% identity) was observed. Its amino acid sequence showed two distinct N- and C-terminal domains (148 and 130 aa, respectively) connected by a peptide linker (63 aa). The galectin contains two consensus WG-E-R/K motifs thought to play an essential role in sugar-binding, indicating that this lectin is a member of the tandem-repeat type galectins which have not been identified in fish. The 1.6 kDa mRNA of the lectin was found by Northern blot analyses to be widely expressed in the spleen, head kidney, thymus, peritoneal exudate cells, ovary, gills and heart. Southern blot analyses with the probe for C-terminal of the lectin showed the existence of two hybridising genes. These results suggest that rainbow trout has at least one tandem-repeat type galectin as well as proto type galectin. PMID- 11394690 TI - Cloning and expression of a putative common cytokine receptor gamma chain (gammaC) gene in rainbow trout (Oncorhynchus mykiss). AB - A full length cDNA of a putative common cytokine receptor gamma chain (gammaC) gene of rainbow trout (Oncorhynchus mykiss) has been cloned and sequenced. The contiguous cDNA contained 2291 nucleotides, consisting of an ORF of 1029 bp, with a 72 bp 5' UTR and a 1190 bp 3' UTR. The coding region showed 44-46% identity to mammalian gammaC genes. The ORF translated into a 343 amino acid protein, with some 28-30% amino acid identity to the coding region of mammalian sequences. A predicted signal peptide and transmembrane domain were identified, giving a 206 amino acid extracellular domain and a 98 amino acid intracellular domain in the trout molecule. Five potential glycosylation sites were present in the extracellular domain, as were six conserved cysteine residues and the W-S-X-W-S motif typical of haemopoietin receptors. One of the most interesting differences between the trout and mammalian sequences was the lack of tyrosines in the trout intracellular domain. RT-PCR studies revealed a wide tissue distribution of gammaC expression, with detectable transcript in blood, spleen, gill, kidney, brain and liver. Low levels of gammaC transcript were detectable in unstimulated macrophage cultures and expression was increased by stimulation of the cells with recombinant trout interleukin-1beta (IL-1beta) or LPS. Similarly, in the RTG cell line which exhibited even lower level constitutive expression, stimulation with IL-1beta increased gammaC transcript levels but LPS had no effect. PMID- 11394691 TI - Tumor necrosis factor alpha (TNFalpha)-like factor produced by macrophages in rainbow trout, Oncorhynchus mykiss. AB - The presence of a Tumor Necrosis Factor alpha (TNFalpha)-like molecule has been suggested in fish by biological assays and biological and antigenic cross reactivities with human TNFalpha. In the present study, whether rainbow trout macrophages produce TNFalpha was examined. Murine recombinant TNFalpha (m rTNFalpha) was used as the standard mammalian TNFalpha. The supernatants were harvested from trout macrophage culture stimulated with lipopolysaccharide (LPS) and then passed through a Polymyxin B column to remove LPS. Results show that trout macrophage culture supernatants exhibit TNF-like activities. The supernatants significantly enhanced neutrophil migration and macrophage respiratory burst activity as assessed by NBT reduction test. The supernatants were also highly cytotoxic to murine L929 cells, which are known to be sensitive to mammalian TNFalpha. The biological activities of TNF in the trout macrophage culture supernatant was determined as 2.6 U ml(-1) in the presence of actinomycin D. This indicates biological cross-reactivity of trout TNFalpha-like factor on mammalian cells. Moreover, these activities were inhibited by a rabbit anti mTNFalpha antibody. These results suggest that rainbow trout macrophages produce a TNFalpha-like factor that is similar to the mammalian TNFalpha in functions. PMID- 11394692 TI - Presence of IgM in cutaneous mucus, but not in gut mucus of Atlantic salmon, Salmo salar. Serum IgM is rapidly degraded when added to gut mucus. AB - In the first part of this study, cutaneous mucus of Atlantic salmon (Salmo salar) was shown to contain IgM, i.e. molecules composed of approximately 72 and 27 kDa subunits and reactive with polyclonal antisera and monoclonal antibodies made against serum IgM. Attempts to detect IgM-like molecules in gut mucus were negative, indicating the IgM is present, at best, in very small amounts. The degradation of serum IgM in mucosal secretions was examined in the second part of this study. Purified IgM from serum was rapidly digested in gut mucus at 4 degrees C. Intermediate 58, 52, 38, 35, 33 and 18 kDa breakdown fragments appeared when analysed in immunoblots. The transient fragments were further degraded to small fragments. HPLC analysis showed that only half of the added serum IgM was intact after 30 min of digestion, and after 4 h intact IgM could not be detected. Serum IgM was not degraded in cutaneous mucus, even after 17 h of incubation. PMID- 11394693 TI - Flow cytometric analysis of European flat oyster, Ostrea edulis, haemocytes using a monoclonal antibody specific for granulocytes. AB - The importance of haemocytes in mollusc defence mechanisms can be inferred from their functions. They participate in pathogen elimination by phagocytosis (Cheng, 1981; Fisher, 1986). Hydrolytic enzymes and cytotoxic molecules produced by haemocytes contribute to the destruction of pathogenic organisms (Cheng, 1983; Leippe & Renwrantz, 1988; Charlet et al., 1996; Hubert et al., 1996; Roch et al., 1996). Haemocytes may also be involved in immunity modulation by the production of cytokines and neuropeptides (Hughes et al., 1990; Stefano et al., 1991; Ottaviani et al., 1996). As a result, the literature dealing with bivalve haemocyte studies has increased during the last two decades. Most of these publications use microscopy for morphological analysis (Seiler & Morse, 1988; Auffret, 1989; Hine & Wesney, 1994; Giamberini et al., 1996; Carballal et al., 1997; Lopez et al., 1997; Nakayama et al., 1997), and functional analysis (e.g. phagocytosis) (Hinsch & Hunte, 1990; Tripp, 1992; Mourton et al., 1992; Fryer & Bayne, 1996; Mortensen & Glette, 1996). Flow cytometry represents a rapid technique applicable to both morphological and functional studies of cells in suspension. While the measurements based on autofluorescence provide information on cell morphology, the analyses with fluorescent markers including labelled antibodies, offer data on phenotyping and cell functions. As a result, its application has greatly contributed to the investigation of immunocyte functions and differentiation in vertebrates (Stewart et al., 1986; Rothe & Valet, 1988; Ashmore et al., 1989; Koumans-van Diepen et al., 1994; Rombout et al., 1996; Caruso et al., 1997). Some authors studied oyster haemocyte populations by flow cytometry based on cellular autofluorescence (Friedl et al., 1988; Fisher & Ford, 1988; Ford et al., 1994). However, no analysis using specific monoclonal antibodies has been reported to date. In this study, a protocol for studying European flat oyster, Ostrea edulis, haemocytes by flow cytometry using a monoclonal antibody specific for granulocytes and an indirect immunofluorescence technique have been developed. European flat oysters, Ostrea edulis, 7-9 cm in shell length were obtained from shellfish farms in Marenne Oleron bay (Charente Maritime, France) on the French Atlantic coast. All individuals were purchased just before each experiment and processed without any previous treatment. PMID- 11394695 TI - The role of guidewire exchange in the treatment of hemodialysis catheter related bloodstream infection. PMID- 11394694 TI - Molecular cloning of carp (Cyprinus carpio) C-type lectin and pentraxin by use of suppression subtractive hybridisation. PMID- 11394696 TI - The role of nandrolone decanoate in patients with end stage renal disease in the erythropoietin era. PMID- 11394697 TI - Phosphatemic control during acute renal failure: intermittent hemodialysis versus continuous hemodiafiltration. AB - BACKGROUND: Achieving "adequacy of dialysis" includes the maintenance of normal serum phosphate concentrations and is an important therapeutic goal in the treatment of acute renal failure (ARF). It is unknown whether this goal is best achieved with intermittent or continuous renal replacement therapy. METHODS: We compared the effects of continuous veno-venous hemodiafiltration (CVVHDF) and intermittent hemodialysis (IHD) on serum phosphate concentrations using daily morning blood tests in 88 consecutive intensive care patients half of which were treated with IHD and half with CRRT RESULTS: Mean patient age was 54+/-14 years for IHD and 60+/-14 years for CVVHDF (NS). However, patients who received CVVHDF were more critically ill (mean APACHE II scores: 24.4+/-5.1 for IHD vs. 29.2+/ 5.7 for CVVHDF, p<0.003). Before treatment, the serum phosphate concentration was 2.04+/-0.16 mmoll L for IHD and 1.96+/-0.17 mmoll L for CVVHDF (NS), with abnormal values in 79.4% of IHD patients and in 64.8% of CVVHDF patients (NS). During treatment, CVVHDF induced a greater reduction in serum phosphate (p=0.02) during the first 48 hours and conferred superior subsequent control of hyperphosphatemia (achieved in 64.6% of observations during CVVHDF vs. 41.8% during IHD; p<0.0001). The serum phosphate concentration was also more likely to be within the normal range during CVVHDF (55.3% vs.36.2%; p<0.0001). There was a trend toward more frequent hypophosphatemia (9.3% vs. 5.6%; P<0.1) during CVVHDF CONCLUSIONS: Abnormal serum phosphate concentrations are frequent in ARF patients before and during renal replacement, however, normalization of phosphatemia is achieved more frequently with CVVHDF. PMID- 11394698 TI - Pre and interdialytic acid-base balance in hemodialysis patients. AB - In an observational study, nine hemodialysis patients using 35 mmol/l bicarbonate dialysate were studied over a 44-hour interdialytic interval. Serum bicarbonate was measured at regular intervals at home and the mean time averaged concentration was 27.0+/-1.2 mmol/l. Seven of the nine patients showed a slow linear decline in bicarbonate whilst in two patients levels were unchanged. In 8 of 9 patients the average of the post and predialysis bicarbonate accurately predicted the time-averaged (area under the curve) bicarbonate concentration. In addition, predialysis serum bicarbonate was measured in 46 patients after both a 2 and 3-day interdialytic interval. The serum bicarbonate was significantly lower after the 3-day interdialytic interval (3-day, 22.1+/-0.6 vs 2-day, 23.0+/-0.5 mmol/l, p<0.05). The results from this study emphasize the importance of standardization of bicarbonate measurement in order to avoid spurious acidosis. PMID- 11394699 TI - Excessive weight gain during peritoneal dialysis. AB - The authors carried out a retrospective chart review in 114 patients treated for at least two years at the Toronto Western Hospital Peritoneal Dialysis Unit and identified eight, who gained an "excessive" amount of weight equal to or greater than 10 kg of their initial weight. These patients had gained an average of 13.1 kg over the preceding two years. They are mostly males and their average age is 51 years. They are well-nourished normotenseive nondiabetics with mostly normal cardiac function. They are adequately dialyzed (per KT/V urea), have little residual renal function and typically have peritoneal membranes characterized by high average transport. According to BIA analysis, this weight gain was likely due to an increase in fat mass accompanied by a trend toward decreasing body-cell mass. This weight gain may be due to increased caloric intake secondary to dialysate glucose absorption in the setting of high average (peritoneal membrane) transport. Such excessive weight gain also may occur if these patients have polymorphism of the UCP-2 gene, which can alter metabolic rate. PMID- 11394700 TI - Renal clearances in continuous ambulatory peritoneal dialysis: differences between diabetic and non-diabetic subjects. AB - We analyzed the effect of diabetes on the decline of residual renal function during the course of CAPD in a cross-sectional study including 105 diabetic subjects (41 women) who had 207 clearance studies and 125 non-diabetic subjects (50 women, 265 clearance studies). CAPD duration was 11.5+/-10.5 months in the diabetic group (DG) and 16.8+/-18.6 months in the non-diabetic group (NDG, P < 0.001). The DG had lower urine volume than the NDG (0.52+/-0.46 vs 0.61+/-0.50 L/24-h, P < 0.05), while urine-to-plasma concentration ratio was higher in the DG for creatinine (13.5+/-9.4 vs 11.5+/-11.0, P <0.05) and did not differ for urea. Weekly renal Kt/V urea (DG 0.51+/-0.57, NDG 0.53+/-0.49) and Ccr (DG 31.0+/-28.7 NDG 29.3+/-26.5 L/1.73 m2) did not differ. The slopes of the regressions of CAPD duration on renal clearances did not differ. These regressions allowed estimates of the time, from the onset of CAPD, at which renal clearances become negligible. These estimates differed for both urea clearance (DG 35.3, NDG 50.5 months) and creatinine clearance (DG 43.2, NDG 57.6 months). The slope of the regression of renal urea clearance on renal creatinine clearance was steeper in the DG, suggesting a higher renal creatinine clearance in the DG than in the NDG when renal urea clearance is the same in the two groups. Subtle differences in the rate of decline of renal function can be detected between diabetic and non diabetic subjects on CAPD by detailed statistical analysis. These findings are supportive of the studies which have identified diabetes mellitus as a predictor of loss of residual renal function during the course of CAPD. In addition, the relationship between the renal urea and creatinine clearances differs between diabetic and nondiabetic subjects on CAPD. Therefore, the dose of CAPD required for adequate total clearances may differ between diabetic and non-diabetic subjects. PMID- 11394701 TI - A comparative prospective study on the use of low concentrate citrate lock versus heparin lock in permanent dialysis catheters. AB - We prospectively evaluated the efficacy and safety of a low concentrate citrate lock versus heparin lock in permanent single lumen hemodialysis catheters. The frequency of clot formation, complete catheter occlusion, flow problems and the use of urokinase as well as catheter infection episodes were monitored during 1370 dialysis sessions in 19 patients, randomised in two study groups. There was a significantly higher number of dialysis sessions with clot formation in the citrate group but regarding the need for urokinase bolus or infusion, complete obstruction of the catheter or local infections, there were no statistically significant differences between groups. The higher incidence of clotting in the citrate locked catheters had no repercussion on dialysis efficiency, effective blood flow or on the use of thrombolytic therapy. We found that low concentrate citrate is as safe as heparin for long-term interdialytic anticoagulation of permanent single lumen hemodialysis catheters but is more efficient from a pharmaco-economic viewpoint. PMID- 11394702 TI - Genes of major histocompatibility complex class II influence chronic C hepatitis treatment with interferon in hemodialysis patients. AB - The prevalence of anti-HCV among patients on hemodialysis is consistently higher than in the general population, indicating that patients on hemodialysis programs are at risk of acquiring HCV infection. The response to interferon alpha 2b (IFN alpha 2b) therapy in chronic C hepatitis depends on viral and host factors. We treated 22 chronic C hepatitis uremic patients with IFN -alpha 2b (3 MU three times a week) and compared interferon responsive and unresponsive patients with regard to HLA II genes. HLA II genes were investigated by PCR-SSP low resolution, anti-HCV with ELISA II and HCV-RNA with reverse transcriptase "nested" PCR. FINDINGS: HLA DRB1*13 is 50% positive in the non-responder group (four women, four men, mean age; 28.8+/-11.9 years) and 7% in the responder group (five women, nine men, mean age; 32.2+/-7.8 years) (p<0.05). There was no difference with respect to HLA genes between controls (six women, eight men, mean age; 29.5+/ 12.8 years) and patients (nine women, 13 men, mean age; 31.0+/-9.3 years) (HLA DRB1*13 is 28% and 22% positive, respectively). We conclude that major histocompatibility complex class II genes influence the outcome of chronic C hepatitis treatment with IFN -alpha 2b. PMID- 11394703 TI - Isolated hemoperfused heart model of slaughterhouse pigs. AB - A model of hemoperfused slaughterhouse pighearts is described providing a wide range of applications which leads to a reduction in animal experiments. The size of a pigheart, heart rate, coronary perfusion, metabolism, etc. are more comparable to conditions in patients than those in hearts of small laboratory animals. Global heart function can be assessed either by measuring stroke volume, ejection fraction, Emax etc. in the working model or by measuring intraventricular pressure with balloon catheters in the isovolumetric model. Regional cardiac function can be measured by sonomicrometry and ischemic and non ischemic areas can be compared. Local metabolic changes are measurable as well with microdialysis. Cardiac function can be kept on any given functional level by infusion of norepinephrine in spite of the fact that functional parameters are lower without adrenergic drive in vitro than in vivo. Stable heart function can be maintained for several hours with only 500 to 1000 ml of blood because the blood is permanently regenerated by a special dialysis system. This model can be applied in many research projects dealing with reperfusion injuries, inotropic, antiarrhythmic or arrhythmogenic effects of certain drugs, immunological rejection, evaluation of imaging systems (NMR, echocardiography etc.) or cardiac assist devices. PMID- 11394704 TI - Lactate dehydrogenase (LD) pattern in serum of calves with a total artificial heart (TAH). AB - Serum lactate dehydrogenase (LD) patterns of 16 calves with total artificial heart (TAH) were studied to identify a mean LD pattern in their serum (survival 51-293 days). Evaluations were made by the sum of vectors method (SV). The mean LD pattern was determined from serum LD patterns, interval days of 33 - 49 with TAH. Serum LD patterns of control and mean LD pattern differed significantly (n = 12, P < 0.05) in single isoenzymes and resultant vectors as the representatives of LD patterns. The different tissues with an increased risk of being affected were identified by evaluation of LD patterns in the serum of calves within term without perceptible acute damage and in the terminal stage of experiments. Such identified organs suggest that the organism under study is out of balance and specific treatment is needed. PMID- 11394705 TI - Standard 19mm St Jude aortic valves in patients with body surface less than 1.7m2. AB - Although new models of bileaflet valves with improved orifice have been devised, aortic valve replacement with 19mm prostheses still raises concerns about long term effects of residual transprosthetic gradient. We reviewed our experience with 19 mm standard model St Jude prostheses in 68 patients operated on between January 1983 and December 1995. Clinical late assessment was performed to evaluate the incidence of valve related complications. Postoperative echocardiography was performed to evaluate hemodynamic performance of the prostheses. Mean body surface area was 1.66+/-0.14 m2. Late postoperative peak gradient was 53.85+/-7.16 mmHg; mean gradient was 34.80+/-5.55 mmHg; effective orifice area was 1.93+/-0.05 cm2. Thirteen-year actuarial survival has been 90.89+/-0.6%; thirteen-year freedom from embolism 89.41+/-0.7% and freedom from hemorrhage 98.25+/-0.02%. No case of prosthetic endocarditis, thrombosis, or reoperation was observed during follow-up. PMID- 11394706 TI - Endothelialization of PTFE vascular grafts under flow induces significant cell changes. AB - BACKGROUND: To minimize thrombogeneity of small diameter PTFE grafts they are usually coated in vitro with endothelial cells under static culture conditions. The disadvantage of this technique is that a cell layer is formed that fails to withstand shear stress typical in normal blood flow. METHOD: Since the in vivo functional and structural status of endothelial cells correlates with the applied shear stress, we developed a computer-controlled perfusion system to seed and culture cells on PTFE-grafts up to a confluent monolayer under the influence of increasing shear stress. The confluence of endothelial coating was defined by immunohistological staining of cross sections, and by upper light microscopy of flattened graft samples. In addition, the expression of fibronectin as an important adhesion molecule was estimated. RESULTS AND DISCUSSION: The application of pulsatile shear stress (6.6 dyn/cm2, 5 min) to grafts endothelialized under perfusion (n = 7) did not lead to a disruption of the confluent cell layer. In contrast, a 5 min long shear stress of 3 dyn/cm2 was sufficient to wash more than 50% of cells off the PTFE-graft cultured under static conditions (n = 6). The perfusion cultures showed a significantly higher proliferation rate in comparison with static cultures. This effect was reproducibile in both serum-containing and serum-free culture media. The expression of fibronectin by endothelial cells was significantly higher in the perfused graft compared to the static one. These results suggest the practicability of endothelialized PTFE vascular grafts, preconditioned to shear rates similar to the in vivo situation, as an alternative bypass material in cardiac surgery. PMID- 11394707 TI - A new semi-automatic endothelial cell seeding technique for biological prosthetic heart valves. AB - BACKGROUND: Until today, tissue heart valve prostheses have been made with biological dead porcine or bovine tissue. However, the durability of this tissue is limited due to degeneration and calcification. Surface seeding with vital human endothelial cells (EC) could improve valve durability and bio compatibility. A new seeding technique that includes a newly developed special seeding device is presented here. METHODS: The aortic valve, including a cylinder of the aortic root, was prepared from a fresh porcine heart taken from the slaughterhouse. Porcine endothelial cells were removed by surface treatment with chemical detergent solutions. A new seeding device with an integrated CO2 incubator was designed. The device is composed of: the seeding chamber (SC), the rotation unit (RU), and the Control Unit (CU). The porcine aortic root cylinder with the valve leaflets is placed into the SC. A matrix of fibronectin is applied to the acellular valve. The SC is then filled with the endothelial cells suspended in modified Dulbecco's eagle medium (DMEM). Under cell culture conditions, the endothelial cell seeding of the tissue valve is established by rotating the valve around two orthogonal axes simultaneously and independently. This is done following the software controlled preset parameters. RESULTS: Using initial endothelial cell seeding concentrations of 6x10(6) endothelial cells/ml DMEM, it was possible to achieve a seeding efficiency of 80-85% within 3-4 hrs. Cell viability tests proved that 90-95% of the seeded endothelial cells are vital after the seeding procedure. CONCLUSIONS: This new seeding technique allows the complex warped surface of a tissue heart valve to be covered with vital endothelial cells to form a confluent endothelial cell monolayer. PMID- 11394708 TI - Optical visualization of microsomal degranulation by a carcinogen. AB - We report here for the first time the optical visualization of microsomal degranulation induced by a carcinogen (p-dimethylaminoazobenzene), employing analytical ultracentrifugation. Our observations demonstrate that electrophiles of a carcinogen can disrupt ribosome-membrane interaction in rough microsomes by their attacks on nucleophilic components of the reticular membrane-ribosome complex involved in protein synthesis for export from the cytosol. Lack of exported proteins can adversely affect signal transduction across plasma membrane, possibly leading to carcinogenesis. PMID- 11394709 TI - Prevalence of human papillomavirus infection among prostitutes in Calcutta. AB - We investigated the presence of human papilloma virus (HPV) 16/18 and herpes simplex virus-1,-2 (HSV-1,-2) infections in buccal mucosal cells of prostitutes of Calcutta, India, by in situ DNA hybridization and immunocytochemical technique. In some of them, we also examined the prevalence of viral infections in uterine cervical cells. The women were also tested for venereal disease research laboratory antigen (VDRL) positivity. Oral infections with HSV-1, -2 and HPV 16/18 were detected in 24.6%, 11.6%, and 29%, respectively. Similar cervical infections were found in 0%, 44%, and 62.9% of the cases studied. HPV coinfection was significantly higher (p < 0.01) in both the oral and cervical cells that had HSV infection. The cervical HPV infections were not markedly higher among the VDRL-positive prostitutes (p > 0.05). The HPV or HSV infection rates showed no remarkable variation between the two age groups of the prostitutes studied (p > 0.05). Cytological abnormalities were more pronounced in the cervix of women having concurrent infection with HPV and HSV-2 rather than in the presence of infection with either of the two. This preliminary investigation, the first of its kind in India, requires a larger study to understand the significance of the association of different sexually transmitted agents including HPV (apart from HIV) as risk factors for pathologic lesions. PMID- 11394710 TI - Sister chromatid exchanges and micronuclei in lymphocytes of operating room personnel occupationally exposed to enfluorane and nitrous oxide. AB - The objective of this article is to assess whether occupational exposure to anesthetics increases genotoxic risk. We investigated two cytogenetic biomarkers, sister chromatid exchanges (SCE) and micronuclei (MN), in the peripheral blood lymphocytes of 46 anesthesiologists (24 men), working in operating rooms and mostly exposed to enfluorane and nitrous oxide, and 66 controls (35 men), not exposed to chemicals and living in the same area. Contrary to what was expected, a lower frequency of SCE was found in male anesthesiologists than in controls. Smoking status was found to be positively associated with SCE frequency in each group, while no relation to age was evident. On the contrary, MN frequency was significantly higher in female, but not male, anesthesiologists than in controls. Age and smoking status did not modify the association. No relationship between MN frequency and duration of employment was found in anesthesiologists. Smoking status and mean number of cigarettes smoked per day in smokers were not associated with MN frequency in either anesthesiologists or in controls. MN analysis seems to be a sensitive index of possible genotoxic effects of occupational exposure to anesthesiologists, and women appear to be more susceptible to these effects than men. PMID- 11394711 TI - Transplacental carcinogenic potential of the carbamate fungicide mancozeb. AB - We evaluated the effects of mancozeb (Dithane M4-5), a protective carbamate fungicide, on transplacental carcinogenesis in Swiss albino mice. Mancozeb, a polymeric complex of ethylene bis (dithiocarbamate) manganese with zinc salt, is reported to possess carcinogenic and cocarcinogenic activity in various tumor models. In the present study, pregnant Swiss albino mice were administered mancozeb intraperitoneally on the 14th day of gestation. The first filial generation (F1 progeny) was promoted with a well-known tumor promoter 12-o tetradecanoyl phorbol-13-acetate (TPA). The results revealed a significantly high tumor incidence (72%) in the F1 progeny of the animals initiated with mancozeb or a well known carcinogen 7,12-dimethyl benzanthracene (DMBA) and promoted with TPA in comparison to animals that were either from mothers given only the vehicle (DMSO) and promoted with TPA in F1 progeny or not promoted with TPA in F1 progeny. No significantly higher tumor incidence was observed in any other experimental groups. These results suggest that mancozeb or its metabolites are capable of crossing the placental barrier and can exert DNA damage and tumor initiating consequences in the fetal cells that, after promotion with TPA, get converted into neoplastic cells. PMID- 11394712 TI - The effect of coenzyme Q10 on blood ascorbic acid, vitamin E, an lipid peroxide in chronic cadmium intoxication. AB - The aim of our study was to investigate the possible protective role of coenzyme Q10 (CoQ10) administration on ascorbic acid (AsA), vitamin E (vit E), and lipid peroxide (LP) concentrations in the blood of rats chronically treated with cadmium. Results were compared to those obtained in control animals, as well as to those obtained in animals treated with olive oil. Compared to that of the control animals, the AsA concentration was significantly increased in rats treated with CoQ10 and olive oil, whereas vit E concentration was significantly increased in animals treated with cadmium, CoQ10, or cadmium + CoQ10. A significant decrease in LP concentration was noted in animals treated with cadmium or with cadmium + CoQ10o, whereas a significant increase was seen in animals treated with olive oil. Compared to that of the animals treated with olive oil, the ascorbic acid concentration was significantly decreased in rats treated with cadmium or with cadmium + CoQ10, whereas vit E concentration was significantly increased in animals treated with cadmium, CoQ10, or cadmium + CoQ10. LP concentration was significantly decreased in rats treated with cadmium, CoQ10, or cadmium + CoQ10. Our study showed that CoQ10 administration in rats chronically exposed to exogenous cadmium exerts beneficial effects on the nonenzymatic components of the antioxidant defense system, such as AsA and vit E, resulting in a decreased concentration of LP in the blood. PMID- 11394713 TI - Influence of caffeine on allyl alcohol-induced hepatotoxicity in rats. I. In vivo study. AB - Cotreatment of rats with a low hepatotoxic dose (30.7 mg/kg, i.p.) of allyl alcohol (AA) and a higher, but nontoxic, dose (150 mg/kg, oral) of caffeine (CF) potentiated the hepatotoxicity of AA. This was verified by significantly higher levels of plasma alanine aminotransferase (ALT) activity and histopathologically greater severity of lesions in the periportal hepatocytes than those due to AA alone. Treatment of rats with 4-methylpyrazole (4-MP) (0.5 mmol/kg, i.p.) (an inhibitor liver alcohol dehydrogenase) for 30 minutes, followed by similar cotreatment with AA and CF, completely prevented the elevation of plasma levels of ALT and histological damage induced by cotreatment with CF and AA 24 hours following their administration. Severe liver damage induced by cotreatment with CF and AA was further, markedly enhanced by phenobarbital pretreatment (80 mg/kg, i.p., 3 days). Thus, extensive necrosis of periportal hepatocytes was noted, as well as edema and accumulation of inflammatory cells in the necrotic foci caused by such pretreatment. The depression of hepatic nonprotein sulfhydryls resulting from CF plus AA was much more severe than that caused by AA or CF alone and appeared as early as 30 minutes after administration. However, much less marked depletion of protein thiols was observed following similar treatments. Significant increase in lipid peroxidation (as measured by melondialdehyde [MDA] formation) was also observed in rat liver but only 24 hours after administration. The production ofMDA in the rat liver was significantly higher after administration of AA plus CF than after administration of AA alone. Pretreatment of rats with phenobarbital further significantly enhanced the formation of 2,4 dinitrophenylhydrazine (DNP)-reactive metabolite(s) (measured as DNP-acrolein adduct equivalents) in rat liver induced by AA (30.7 mg/kg) plus CF (150 mg/kg) within 1 hour following such treatment. Cotreatment with AA and a higher dose of CF resulted in significantly higher excretion of urinary thioethers or mercapturic acids than in rats treated with AA alone. Thus, these data suggest that an increased bioactivation pathway of acrolein involving a P450 mixed function oxidase system caused by CF may be involved in such potentiating effects of CF on AA-induced hepatotoxicity in rats. PMID- 11394714 TI - Caffeine potentiation of allyl alcohol-induced hepatotoxicity. II. In vitro study. AB - We examined the effects of caffeine (C) on allyl alcohol (AA)- and acrolein (A) induced hepatotoxicity on freshly-isolated, rat hepatocytes obtained from livers of adult, male, Sprague-Dawley rats. Isolated rat hepatocytes in suspension were incubated in each test with one of the following: 0, 1.0, or 2.5 mM of AA alone; or with 0, 2.5, or 5 mM of C alone; or a combination of AA and C at the same range of concentrations as used alone, for 15, 30, 60, 90, and 120 minutes at 37 degrees C. A dose- and time-dependent potentiation of cytotoxicity as measured by cellular viability (using trypan blue exclusion) were observed. The AA (2.5 mM) induced lactate dehydrogenase (LDH) leakage observed after 60 minutes incubation was completely prevented when pretreated for 15 minutes with 4-methylpyrazole (MP) (0.5 mM). Such pretreatment, even with a double dose of 4-MP, only partially, and not significantly, prevented LDH leakage when the hepatocytes were incubated with a mixture of 2.5 mM AA and 5 mM C. The depletion of hepatocyte nonprotein sulfhydryl (NPSH) content caused by AA was further enhanced in the presence of C, as early as 15 minutes after their exposure. The AA-induced increase in lipid peroxidation was also potentiated by C; however, potentiation started later, and only after sufficient depletion of NPSH (mostly glutathione) occurred resulting from the presence of AA plus C. A significant loss of protein sulfhydryls in rat hepatocytes could be noted following a 60-minute incubation period with either AA (1 mM) or AA (1 mM) plus C (5 mM). Similarly, C produced a dose-and time-dependent potentiation of A-induced liver cytotoxicity, which was preceded by severe loss of NPSH content within 15 minutes of exposure, whereas the potentiation of lipid peroxidation (LPO) resulting from A plus C was found to be a relatively late event, as with AA plus C. Furthermore, combined treatment with AA and C produced a significantly higher cytotoxicity (as measured by cellular viability) than that due to the combined treatment with A plus C based on equimolar concentration. These results suggest that two increased bioactivation pathways of AA involving the P-450 mixed-function oxidase system resulting from C may be involved in the potentiation of AA hepatotoxicity. PMID- 11394715 TI - Oxidative mechanisms in the toxicity of chromium and cadmium ions. AB - Chromium and cadmium are widely used industrial chemicals. The toxicities associated with both metal ions are well known. However, less information is available concerning the mechanisms of toxicity. The results of in vitro and in vivo studies demonstrate that both cations induce an oxidative stress that results in oxidative deterioration of biological macromolecules. However, different mechanisms are involved in the production of oxidative stress by chromium and cadmium. Chromium undergoes redox cycling, while cadmium depletes glutathione and protein-bound sulfhydryl groups, resulting in enhanced production of reactive oxygen species such as superoxide ion, hydroxyl radicals, and hydrogen peroxide. These reactive oxygen species result in increased lipid peroxidation, enhanced excretion of urinary lipid metabolites, modulation of intracellular oxidized states, DNA damage, membrane damage, altered gene expression, and apoptosis. Enhanced production of nuclear factor-kappaB and activation of protein kinase C occur. Furthermore, the p53 tumor suppressor gene is involved in the cascade of events associated with the toxicities of these cations. In summary, the results clearly indicate that although different mechanisms lead to the production of reactive oxygen species by chromium and cadmium, similar subsequent mechanisms and types of oxidative tissue damage are involved in the overall toxicities. PMID- 11394716 TI - Bioactive anthocyanins detected in human urine after ingestion of blackcurrant juice. AB - Anthocyanins are a group of very efficient bioactive compounds that are widely distributed in plant food. Several fruits (blackcurrant, blackberry, red grape) and some vegetables (eggplant, onion, red radish) are rich sources of these natural pigments. Extracts of some of them are used as food colorants as well as components of pharmaceutical preparations and functional foods. Anthocyanins, through their ability to inhibit radical reactions, are considered to exert several protective effects in the human body. Until now there has been only a small amount of data available on their capability, in intact or metabolized form, to reach the systemic circulation of humans. The present study was designed to determine the potential bioavailability in humans of the most important anthocyanins of blackcurrants: delphinidine-3-glucoside, delphinidine-3 rutinoside, cyanidine-3-glucoside, and cyanidine-3-rutinoside. Urinary samples from 4 healthy volunteers (2 women and 2 men) were collected before (baseline) and over a period of 5 hours with intervals of 30 minutes after the ingestion of 200 mL of blackcurrant juice (containing 153 mg of anthocyanins). Using high performance liquid chromatography (HPLC), it was possible to quantify the 4 main anthocyanins of blackcurrants, excreted unchanged in the urine (0.020-0.050% of the oral doses). We present data on the bioavailability in humans of blackcurrant anthocyanins, which are dietary antioxidants with possible biological effects. PMID- 11394718 TI - Naming rotated pictures and the riddle of object-centred neglect. AB - In this study eight patients with left neglect were asked to name chimerical pictures of objects and animals with different spatial orientation: standard upright position, rotated 180 degrees, rotated 90 degrees to the right, and rotated 90 degrees to the left. All patients showed the typical pattern of egocentric neglect. They omitted the left part of the normally upright pictures and the right part of the inverted stimuli, now falling in the left space. When the pictures were tilted 90 degrees to the right, they reported the two component objects with the same level of accuracy. However, at variance with egocentric neglect, when the chimerical pictures were rotated 90 degrees to the left, the patients omitted the left half of the stimulus more often than the right half. We propose that since in the latter condition the less informative lower part of the pictures was available in the non-neglected space, the patients mentally rotated the perceived stimulus and aligned it with its upright orientation before naming its component parts. In our interpretation, the mental orientation and normalisation of rotated stimuli might underlie all the reported evidence of object-centred neglect for non-orthographic stimuli. PMID- 11394717 TI - Increased TGF-beta1 in the lungs of asbestos-exposed rats and mice: reduced expression in TNF-alpha receptor knockout mice. AB - Inhalation of numerous fibrogenic agents causes interstitial pulmonary fibrosis (IPF) in humans and in a number of animal models. Several of these models provide evidence that certain peptide growth factors (GF) are playing a role in the disease process. Transforming growth factor beta 1 (TGF-beta1) is a potent inducer of extracellular matrix production by mesenchymal cells, and we have shown that this peptide is produced in the lung after asbestos exposure. We used in situ hybridization to demonstrate that the mRNA for TGF-beta1 is rapidly expressed post-exposure at sites of initial asbestos-induced lung injury in both rats and mice. The TGF-beta1 is expressed by bronchiolar-alveolar epithelial cells as well as by mesenchymal cells and lung macrophages in exposed animals. Normal rats and mice express little TGF-beta1, as we have demonstrated previously for PDGF-A and -B, TGF-alpha, and TNF-alpha. TGF-beta1 expression is accompanied by collagen and fibronectin production in asbestos-exposed animals. Most interesting, TGF-beta1 expression is largely absent in the lungs of TNF-alpha receptor knockout mice that fail to develop asbestos-induced IPE We have shown previously that the mRNAs and cognate peptides of PDGF-A and -B and TGF-alpha, but not TNF-alpha, are reduced in the fibrosis-resistant knockout mice. In this article, we show that TGF-beta1 is included in this group of cytokines, supporting the postulate that TNF-alpha is necessary for the expression of other, more downstream growth factors, and the consequent development of idiopathic pulmonary fibrosis (IPF). PMID- 11394719 TI - The influence of misnaming on object recognition: a case of multimodal agnosia. AB - We present a case of multimodal agnosia in the visual and tactile modality due to an infarction in the territory of the left posterior cerebral artery. The patient's ability to recognize objects fluctuated depending on his verbal activity. When he misnamed presented objects, he tended to use them and to draw them in keeping with the wrong name. We submit that the mechanism causing associative agnosia is more dynamic than it was hitherto considered. It originates from the rivalry between top-down central regulation and bottom-up peripheral flow. PMID- 11394720 TI - Pure Kana agraphia as a manifestation of graphemic buffer impairment. AB - We report a left-handed man who demonstrated a pure agraphia limited to words written in Kana characters (syllabograms) following a right putaminal hemorrhage. Writing words in Kanji characters (logograms) was well preserved. His performance in Kana writing was characterized by intact ability to write single syllables, error increase in the second half of words directly proportional to the word length and correct but slow writing of words using kana blocks. Errors were more prominent in Hiragana words than Katakana words which are usually used to transcribe foreign words. Acoustic-grapheme sequencing per se was not impaired as shown by his correct performance in arranging character blocks. These findings suggest selective damage to the graphemic buffer, a module that temporarily maintains the graphemic representation elaborated in previous stages before it is sent to the peripheral systems for its motor realization. PMID- 11394721 TI - Abnormal semantic network for "animals" but not "tools" in patients with Alzheimer's disease. AB - The effects of Alzheimer's disease (AD) on the organization of semantic knowledge (i.e., the semantic network) for living (e.g., animals) and non-living (e.g., tools) categories was examined. Multidimensional scaling and Pathfinder analyses of data from triadic comparison tasks showed that the semantic network for "animals", but not the network for "tools", was abnormal in patients with AD. Specifically, patients with AD tended to use a different primary dimension than control subjects for categorizing animals and their network was characterized by atypical associations between concepts. The differences in the integrity of the AD patients' networks for "animals" and "tools" was not likely to be an artifact of differences in the difficulty in identifying the stimuli in the two categories as all stimuli were identified on simple naming or matching tasks. These findings support the results of previous studies that have shown the presence of category specific semantic deficits in patients with AD. PMID- 11394722 TI - Praxis lateralization: errors in right and left hemisphere stroke. AB - Whereas the representations of skilled movements in most right handers are stored in the left hemisphere, the right hemisphere's contribution to action planning remains unclear. We investigated error patterns in left (LHD) and right hemisphere damaged (RHD) subjects as well as normal control subjects (C) to determine if specific components of action programs may be processed by the right hemisphere or bilaterally represented. We had these subjects perform gestures to verbal command with the ipsilesional limb. Although the LHD group made significantly more qualitative errors than the C and RHD groups, the RHD subjects produced a number of apraxic errors. Specifically, the LHD group produced a wide range of spatiotemporal and conceptual errors for both transitive and intransitive gestures, while the RHD group made specific spatial and temporal errors primarily when performing transitive gestures. These findings support the postulate that the left hemisphere stores the spatiotemporal and conceptual representations of learned skilled movements, while several specific components of action programs, such as external configuration (limb orientation) and timing, may have bihemispheric representations. PMID- 11394723 TI - Lateral preferences among indian school children. AB - In the first study, 718 children from India, aged 4 to 11 years, were observed for their hand preference for ten common unimanual activities. The prevalence of left-handedness was found to be only 3.2 percent, i.e. one-third of that (9.6%) observed in a French study using a similar procedure. The degree (weak to strong) but not the direction of hand preference was found to be related to the children's age, with stronger preference among older children. The factor structure of handedness items was similar in France and India. In the second study, 400 schoolchildren from India, aged 6 to 18 years, were examined for handedness, footedness, eyedness, use of hand in space, and absolute and relative hand skill assessed by a peg-moving task and a dot-filling task. Prevalence of left-handedness was 4.2%. A sex difference was observed for handedness, footedness, use of hand in space. and relative hand skill, with higher proportions of right preferences and higher degree of lateralization (i.e., relative between hands asymmetry) in females. The degree of hand skill asymmetry increased with age. These results are discussed in relation to findings from previous studies in other countries using similar procedures. PMID- 11394724 TI - A pet study of human skill learning: changes in brain activity related to learning an orientation discrimination task. AB - Using 15O-water 3D positron emission tomography we investigated the effect of training in orientation discrimination upon cerebral activity in healthy human adults. When subjects are trained in this discrimination task, they learn the visuo-motor stimulus-response association required by the task and they increase their perceptual abilities in orientation discrimination. The present study was designed to investigate the rCBF modifications related to both these learning processes induced by training in orientation discrimination. PET data were acquired on two separate days (before and after training). Comparing the activation pattern related to orientation discrimination before and after the training period we observed activity decreases located in the left cerebellar cortex, in the right precentral gyrus and bilaterally in the fusiform gyri. The only region showing an activity increase was located in the body of the right caudate nucleus. These findings confirm the role of the neostriatum in skill learning and highlight the importance of mechanisms resulting in cortical and cerebellar neuronal activity decreases in this type of learning. PMID- 11394725 TI - Door but not four and 4 a category specific transcoding deficit in a pure acalculic patient. AB - We report a left parietal damaged, acalculic, non aphasic patient who showed a specific deficit in reading Arabic and spelled-out numerals. Word reading was flawless, while he showed a severe impairment in reading meaningless strings of phonemes (phonological alexia). He also showed a dissociation between the preserved ability to retrieve knowledge about general facts and the impairment in retrieving cardinal, personal and non-personal numerical facts. These findings point to a separate organization in the brain of the numerical domain. PMID- 11394726 TI - Matching treatment to pathophysiology in type 2 diabetes. AB - BACKGROUND: Because type 2 diabetes is a progressive condition, >50% of all patients whose disease is initially controlled with diet and exercise will eventually need single or multiple pharmacologic agents to maintain adequate glycemic control. Although current treatment standards require that combination therapy be instituted only after the failure of monotherapy, the results of the Diabetes Control and Complications Trial and the United Kingdom Prospective Diabetes Study suggest that aggressive initial treatment is crucial to slowing the evolution of long-term complications associated with this disease. OBJECTIVES: This article reviews the diagnosis and classification of type 2 diabetes, describes the multiple defects in glucose metabolism associated with the disease, and discusses the various pharmacologic options for achieving glycemic control in these patients. METHODS: The information in this review was compiled through a search of MEDLINE. Search terms included but were not limited to type 2 diabetes and antihyperglycemic agents. In addition, abstracts were identified using the Web sites of diabetes-related professional organizations. RESULTS: Two pathophysiologic mechanisms, insulin resistance and impaired insulin secretion, are usually present at diagnosis in type 2 diabetes. Several studies have shown that combination therapy with antihyperglycemic agents having different mechanisms of action provides greater efficacy than treatment with single agents. CONCLUSIONS: Current research suggests that early aggressive treatment with combination therapy achieves glycemic control at lower doses and with fewer side effects than monotherapy with either component. PMID- 11394727 TI - A review of omeprazole use in the treatment of acid-related disorders in children. AB - BACKGROUND: Acid peptic disease is a common problem, with a similar prevalence of gastroesophageal reflux disease (GERD) in adults and children. The presentation of GERD in infants and children varies from crying, irritability, or sleep disturbance to feeding difficulties, vomiting, or rumination. Helicobacter pylori (HP)-related diseases and gastric and duodenal ulcers are much more common in adults than in children, who are more likely to have gastritis or duodenitis. However, because HP infection is most likely acquired in childhood, treatment of children with endoscopically documented active HP disease may minimize the potential risk for peptic ulcer or gastric cancer in adulthood, although this is yet to be proved. OBJECTIVE: Omeprazole has been shown to be effective in the treatment of acid-related diseases. This paper reviews the literature on the use and administration of omeprazole for the treatment of GERD, peptic ulcer disease, HP infection, and other acid-related conditions in children. METHODS: Studies were identified through searches of MEDLINE and Science Citation Index for the period 1986 to November 2000, and from the reference lists of identified articles. The search terms used included omeprazole, proton pump inhibitor (PPI), children, pediatrics, routes of administration, GERD, HP infection, esophagitis, and administration. In addition, the manufacturer of omeprazole was asked for relevant unpublished information. RESULTS: Marketed and extemporaneous formulations of omeprazole have been administered to children aged 2 months to 18 years for the treatment of erosive esophagitis, gastric ulcer, duodenal ulcer, HP infection, and related conditions at dosages of 5 to 80 mg/d (0.2-3.5 mg/kg/d) for periods ranging from 14 days to 36 months with a low incidence of adverse effects. The initial dose most consistently reported to heal esophagitis and provide relief of symptoms of GERD appears to be 1 mg/kg per day. CONCLUSIONS: In uncontrolled clinical trials and case reports to date, omeprazole has been effective and well tolerated for the acute and chronic treatment of esophageal and peptic ulcer disease in children, particularly those who had failed to respond to previous treatment with histamine2-receptor antagonists. Should future long-term, controlled clinical trials in children demonstrate safety and efficacy, this PPI is likely to find a place in the armamentarium of pediatric pharmacotherapy. PMID- 11394728 TI - Oxcarbazepine, an antiepileptic agent. AB - BACKGROUND: Epilepsy is a common neurologic condition. Many of the currently approved pharmacologic agents for its treatment are associated with numerous adverse drug reactions and drug interactions. OBJECTIVE: This review describes the pharmacology and therapeutic use of oxcarbazepine, an analogue of the well known antiepileptic agent carbamazepine. METHODS: Articles for review were identified through a search of MEDLINE, International Pharmaceutical Abstracts, and EMBASE for the years 1980 through 2000. The terms used individually and in combination were oxcarbazepine, carbamazepine, epilepsy, and seizures. RESULTS: Oxcarbazepine and its primary metabolite have been effective in animal models of epilepsy that generally predict efficacy in generalized tonic-clonic seizures and partial seizures in humans. The exact mechanism of action of oxcarbazepine is unknown, although as with carbamazepine, it is believed to involve blockade of voltage-gated sodium channels. The pharmacokinetic profile of oxcarbazepine is less complicated than that of carbamazepine, with less metabolism by the cytochrome P450 system, no production of an epoxide metabolite, and lower plasma protein binding. The clinical efficacy and tolerability of oxcarbazepine have been demonstrated in trials in adults, children, and the elderly. In a double blind, randomized, crossover trial in adults, oxcarbazepine 300 mg was associated with a decrease in the mean frequency of tonic seizures (21.4 vs 30.5 seizures during steady-state periods) and tonic-clonic seizures (8.2 vs 10.4) compared with carbamazepine 200 mg (P = 0.05). A multinational, multicenter, double-blind, placebo-controlled, randomized, 28-week trial assessed the efficacy and tolerability of oxcarbazepine at doses of 600, 1200, and 2400 mg as adjunctive therapy in patients with uncontrolled partial seizures. All 3 oxcarbazepine groups demonstrated a reduction in seizure frequency per 28-day period compared with placebo (600 mg, 26% reduction; 1200 mg, 40% reduction; 2400 mg, 50% reduction; placebo, 7.6% reduction; all, P < 0.001). A trial in children assessed the efficacy and toxicity of oxcarbazepine (median dose, 31.4 mg/kg/d) as adjunctive therapy for partial seizures. Patients receiving oxcarbazepine experienced a 35% reduction in seizure frequency, compared with a 9% reduction in the placebo group (P < 0.001). The most common adverse effects associated with oxcarbazepine are related to the central nervous system (eg, dizziness, headache, diplopia, and ataxia) and the gastrointestinal system (eg, nausea and vomiting). Compared with carbamazepine, there is an increased risk of hyponatremia with oxcarbazepine. The frequency and severity of drug interactions are less with oxcarbazepine than with carbamazepine or other antiepileptic agents. CONCLUSIONS: Oxcarbazepine may be considered an appropriate alternative to carbamazepine for the treatment of partial seizures in patients who are unable to tolerate carbamazepine. Its use in nonseizure disorders remains to be examined in large scale clinical trials, and pharmacoeconomic comparisons of oxcarbazepine with other antiepileptic agents, particularly carbamazepine, are needed. PMID- 11394729 TI - Use of changes in symptoms to predict changes in lung function in assessing the response to asthma therapy. AB - BACKGROUND: The majority of adult patients with asthma are managed by primary care providers. Although there is no generally accepted gold standard for the assessment of asthma severity in general practice, treatment decisions and modifications to therapy are strongly influenced by patients' symptoms and history of asthma medication use. OBJECTIVES: The primary goal of this study was to determine whether there is a correlation between changes in asthma symptoms during treatment and changes in lung function, as measured by peak expiratory flow (PEF). A secondary goal was to compare the relative efficacy (in terms of improvement in asthma symptoms and lung function) of 3 commonly used asthma treatments: inhaled fluticasone propionate, inhaled salmeterol xinafoate, and oral zafirlukast. METHODS: This was a retrospective comparison employing regression analyses of asthma symptom and lung function data from 2890 male and female adolescent and adult patients with persistent asthma who were enrolled in 8 randomized, double-blind, double-dummy, parallel-group studies. Data on patients' self-rated symptoms, PEF, supplemental albuterol use, nighttime awakenings, and frequency of asthma exacerbations were used to ascertain whether there was a correlation between changes in asthma symptoms and changes in pulmonary function, and to compare treatment effects. RESULTS: Changes in patients' ratings of asthma symptoms after treatment with study medications showed a strong correlation with changes in lung function. Similarly, changes in lung function were strongly correlated with changes in supplemental beta-agonist use and quality of life. In addition, fluticasone or salmeterol treatment resulted in significantly greater increases in mean morning PEF (P < 0.001), significantly greater decreases in symptom scores (P < or = 0.004), significantly fewer nights with awakenings due to symptoms (P < or = 0.017), and significantly greater reductions in supplemental beta-agonist use (P < 0.001) compared with zafirlukast treatment or placebo. Patients treated with fluticasone or salmeterol also experienced significantly lower rates of asthma exacerbation (3%) during treatment than did those receiving zafirlukast (7%) or placebo (12%) (P < 0.001 and P = 0.015, fluticasone and salmeterol, respectively). CONCLUSION: These findings support the validity of primary care practitioners' basing asthma management decisions on patients' symptoms. PMID- 11394730 TI - Tolerability of antihypertensive drugs in a community-based setting. AB - BACKGROUND: Outside the experimental environment of clinical trials, the tolerability of angiotensin-converting enzyme inhibitors (ACEIs), calcium channel blockers (CCBs), and the angiotensin II antagonist losartan has not been compared. OBJECTIVES: The purpose of this study was to estimate, in current clinical practice, the 3-month cumulative incidence of side effects among first time users of losartan, ACEIs, and CCBs for hypertension. METHODS: We conducted a prospective cohort study through a network of 173 pharmacies across Canada to identify patients with hypertension who were newly prescribed monotherapy with losartan, an ACEI, or a CCB. Individuals were interviewed by telephone 3 times over a 3-month period to determine perceived side effects of the antihypertensive medication prescribed. Data were analyzed using a multivariate logistic regression model. RESULTS: Among the 663 eligible individuals, the 3-month cumulative incidence of perceived side effects was 52.5% (42/80), 60.2% (222/369), and 69.6% (149/214) for those treated with losartan, an ACEI, and a CCB, respectively. After adjustment for sex, age, level of education, number of symptoms perceived the week before entering the study, prior use of antihypertensive drugs, current use of any other drug, drug insurance coverage, and duration of hypertension, the odds of reporting a side effect were significantly higher among patients treated with an ACEI (odds ratio [OR] = 1.78: 95% CI, 1.02-3.12) or a CCB (OR = 2.65; 95% CI, 1.47-4.78) compared with patients treated with losartan. CONCLUSIONS: In a community-based setting, we observed that losartan is better tolerated than ACEIs and CCBs. Given that the occurrence of side effects may contribute to lower adherence to drug treatment, the low incidence of side effects associated with losartan makes it an attractive antihypertensive drug choice. PMID- 11394731 TI - Initiation of nonselective alpha1-antagonist therapy and occurrence of hypotension-related adverse events among men with benign prostatic hyperplasia: a retrospective cohort study. AB - BACKGROUND: Treatment of benign prostatic hyperplasia (BPH) with nonselective alpha1 antagonists such as terazosin, doxazosin, and prazosin results in blood pressure reduction due to vasodilation. OBJECTIVE: Using claims data from a large Medigap plan, we examined the effect of initiating nonselective alpha1-antagonist therapy on the incidence of hypotension-related adverse events likely to be associated with vascular alpha-adrenoreceptor antagonism in patients with BPH. METHODS: Medical and prescription claims data were obtained from the MEDSTAT Group for 53,824 men with a diagnosis code for BPH during the study period (January 1995-December 1997). We examined the rate of possible hypotension related adverse events (diagnosis codes for hypotension, syncope, dizziness, fractures, and other injuries) per 10,000 person-days for men who began therapy with alpha1 antagonists and for a random sample of nonusers, stratified by prior use of other antihypertensive agents. RESULTS: After adjusting for baseline differences in event rates, those who initiated alpha1-antagonist therapy (n = 1564) had a significantly greater increase in hypotension-related adverse-event rates in the 4 months after initiation (vs the 4 months before initiation) than randomly selected nonusers (n = 8641) (increase of 1.82 vs decrease of 0.02 events per 10,000 person-days among those not taking antihypertensive agents; increase of 0.94 vs 0.69 events per 10,000 person-days among those taking other antihypertensive agents; P < 0.01). This increase began earlier and lasted longer among patients taking other antihypertensive agents. Those who discontinued their alpha1 antagonist had a higher rate of hypotensive events at baseline than those who did not (5.09 vs 3.19 events per 10,000 person-days among those using other antihypertensive agents; 3.62 vs 2.27 events per 10,000 person-days among those not using other antihypertensive agents; P < 0.05). CONCLUSIONS: Initiation of nonselective alpha1-antagonist therapy for the treatment of BPH increases the risk of a cluster of clinical events consistent with vascular alpha adrenoreceptor antagonism. This effect is seen during a 4-month period around the initiation date. Prior initiation of other antihypertensive medication increases this effect. Urologists should consult with a patient's primary care physician about use of other antihypertensive agents before initiating nonselective alpha1 antagonist therapy for BPH. PMID- 11394732 TI - Late-term smoking cessation despite initial failure: an evaluation of bupropion sustained release, nicotine patch, combination therapy, and placebo. AB - OBJECTIVE: The purpose of this study was to evaluate the efficacy of long-term use of bupropion sustained release (SR), the nicotine patch, and the combination of these 2 treatments in patients who initially failed treatment. METHODS: This was a post hoc analysis of a multicenter, double-blind, randomized, placebo controlled clinical trial in 893 smokers. Patients were randomly assigned to 9 weeks of treatment with placebo (n = 160), bupropion SR (n = 244), nicotine patch (n = 244), or a combination of nicotine patch and bupropion SR (n = 245). The study was originally designed with a follow-up period of 52 weeks. In this analysis, short-term success was defined as smoking cessation after 14 or 21 days of therapy and long-term success was defined as smoking cessation after >21 days of therapy. Patients who did not achieve short-term success were evaluated for long-term success at week 9 (end of treatment), 6 months, and 1 year after the start of the study. RESULTS: The mean age of the smokers was 44 years. The majority (93%) of patients were white, and 52% were female. The study subjects smoked an average of 27 cigarettes per day. Among the 467 patients who initially failed treatment in the first 3 weeks, treatment with bupropion SR alone and in combination with the nicotine patch produced significant increases in successful smoking cessation rates from weeks 4 to 9 (19% bupropion SR or combination, 7% nicotine patch, 7% placebo), at month 6 (11% bupropion SR, 13% combination, 2% nicotine patch, 3% placebo), and at month 12 (10% bupropion SR, 7% combination, 2% nicotine patch, 1% placebo) (P < 0.05 for bupropion SR and combination vs nicotine patch or placebo). CONCLUSION: Among patients who initially failed treatment, continued therapy with bupropion SR, either alone or in combination with the nicotine patch, resulted in significantly higher short- and long-term smoking cessation rates than treatment with the nicotine patch alone or placebo. PMID- 11394733 TI - A comparison of the effects on saliva output of oxybutynin chloride and tolterodine tartrate. AB - BACKGROUND: Oxybutynin chloride and tolterodine tartrate are anticholinergic agents used to suppress involuntary bladder contractions in urinary incontinence. They act by inhibiting binding of acetylcholine to the muscarinic receptors in the detrusor muscle of the bladder. The same types of muscarinic receptors are found in the salivary glands; thus anticholinergic agents may decrease saliva production and cause dry mouth, a commonly cited reason for discontinuation of therapy. OBJECTIVE: The primary objective of this study was to compare saliva output, which is an objective measure of dry mouth, in subjects taking immediate- or extended-release oxybutynin, tolterodine, or placebo. METHODS: This was a single-site, single-dose, randomized, double-blind, 4-treatment, 4-period crossover study. Subjects were randomly assigned to 1 of 4 treatment sequences that included extended-release oxybutynin 10 mg, tolterodine 2 mg, immediate release oxybutynin 5 mg, and placebo. Saliva output was measured objectively before dosing with each treatment and at 0.5, 1, 2, 3, 4, 6, 8, 10, and 12 hours after dosing. RESULTS: Thirty-six healthy adult volunteers (22 women and 14 men) participated in the study. They ranged in age from 19 to 42 years (mean, 27 years). Thirty-one were white, 3 Asian, and 2 black. There were no significant differences in predose saliva output between the 4 study groups. With placebo, saliva output increased throughout the day. Saliva output was maintained at predose levels throughout the day with extended-release oxybutynin. Two hours after dosing with tolterodine and immediate-release oxybutynin, saliva output decreased nearly 0.5 g in specimens collected over 2 minutes. All 3 active treatments were associated with lower saliva output compared with placebo. Extended-release oxybutynin and tolterodine were similar with respect to area under the saliva concentration-time curve but were associated with significantly greater saliva output than was immediate-release oxybutynin (P < 0.01). There were no serious adverse events (AEs) in this study. AEs were similar between treatments, although the incidence of headache was higher in the active-treatment groups than with placebo. CONCLUSIONS: Objective assessment of saliva output in healthy adult volunteers indicated that extended-release oxybutynin and tolterodine had less impact on saliva output than did conventional immediate release oxybutynin, suggesting that they may yield lower levels of dry mouth. PMID- 11394734 TI - Lansoprazole-based triple therapy versus ranitidine bismuth citrate-based dual therapy in the eradication of Helicobacter pylori in patients with duodenal ulcer: a multicenter, randomized, double-dummy study. AB - BACKGROUND: The optimal treatment regimen for eradication of Helicobacter pylori in patients with duodenal ulcer has yet to be determined. Based on a search of MEDLINE, no studies have been performed comparing a proton pump inhibitor-based triple therapy regimen with a ranitidine bismuth citrate (RBC)-based dual therapy regimen, both containing clarithromycin. OBJECTIVE: This study was undertaken to compare the efficacy of lansoprazole (LAN)-based triple therapy with that of RBC based dual therapy in H pylori-infected patients with duodenal ulcer. METHODS: Patients were randomized to receive either 1 week of triple therapy with LAN 30 mg BID, clarithromycin 500 mg BID, and tinidazole 500 mg BID, followed by 3 weeks of LAN 30 mg BID, or 2 weeks of dual therapy with RBC 400 mg BID plus clarithromycin 500 mg BID, followed by 2 weeks of RBC 400 mg BID. Eradication of H pylori was defined as negative results on both the urease quick test and histologic examination > or =4 weeks after the end of treatment. Duodenal healing and recurrence rates were assessed endoscopically at 8 weeks and 6 months. A per protocol (PP) analysis was conducted for each efficacy end point. Also conducted were an intent-to-treat (ITT) analysis in which patients with missing data were considered failures, and an observed analysis (OBS), which included patients with an evaluable result after treatment, regardless of compliance. RESULTS: One hundred eighty-five patients (126 men, 59 women; age range, 18-76 years; mean age, 43 years) were enrolled and randomized to treatment. In the LAN and RBC groups, respectively, H. pylori eradication rates were 92.6%, 93.1%, and 72.8% versus 78.6%, 77.9%, and 64.5% in the PP (P = 0.02), OBS (P = 0.01), and ITT analyses. The corresponding duodenal ulcer healing rates were 98.6%, 98.7%, and 83.7% versus 90.8%, 91.5%, and 81.7%; these differences were not statistically significant. Side effects were mild, occurring in 20.7% of LAN patients and 17.2% of RBC patients. Ulcer recurred in 2 RBC patients. No difference was observed between treatments in terms of the occurrence of gastritis or improvement of symptoms. CONCLUSION: Based on the results of the PP and OBS analyses, LAN-based triple therapy was superior to RBC-based dual therapy for the eradication of H. pylori in patients with duodenal ulcer. PMID- 11394735 TI - Structure-based 3D QSAR and design of novel acetylcholinesterase inhibitors. AB - The paper describes the construction, validation and application of a structure based 3D QSAR model of novel acetylcholinesterase (AChE) inhibitors. Initial use was made of four X-ray structures of AChE complexed with small, non-specific inhibitors to create a model of the binding of recently developed aminopyridazine derivatives. Combined automated and manual docking methods were applied to dock the co-crystallized inhibitors into the binding pocket. Validation of the modelling process was achieved by comparing the predicted enzyme-bound conformation with the known conformation in the X-ray structure. The successful prediction of the binding conformation of the known inhibitors gave confidence that we could use our model to evaluate the binding conformation of the aminopyridazine compounds. The alignment of 42 aminopyridazine compounds derived by the docking procedure was taken as the basis for a 3D QSAR analysis applying the GRID/GOLPE method. A model of high quality was obtained using the GRID water probe, as confirmed by the cross-validation method (q2LOO = 0.937, q2L50%O = 0.910). The validated model, together with the information obtained from the calculated AChE-inhibitor complexes, were considered for the design of novel compounds. Seven designed inhibitors which were synthesized and tested were shown to be highly active. After performing our modelling study the X-ray structure of AChE complexed with donepezil, an inhibitor structurally related to the developed aminopyirdazines, has been made available. The good agreement found between the predicted binding conformation of the aminopyridazines and the one observed for donepezil in the crystal structure further supports our developed model. PMID- 11394736 TI - DOCK 4.0: search strategies for automated molecular docking of flexible molecule databases. AB - In this paper we describe the search strategies developed for docking flexible molecules to macomolecular sites that are incorporated into the widely distributed DOCK software, version 4.0. The search strategies include incremental construction and random conformation search and utilize the existing Coulombic and Lennard-Jones grid-based scoring function. The incremental construction strategy is tested with a panel of 15 crystallographic testcases, created from 12 unique complexes whose ligands vary in size and flexibility. For all testcases, at least one docked position is generated within 2 A of the crystallographic position. For 7 of 15 testcases, the top scoring position is also within 2 A of the crystallographic position. The algorithm is fast enough to successfully dock a few testcases within seconds and most within 100 s. The incremental construction and the random search strategy are evaluated as database docking techniques with a database of 51 molecules docked to two of the crystallographic testcases. Incremental construction outperforms random search and is fast enough to reliably rank the database of compounds within 15 s per molecule on an SGI R10000 cpu. PMID- 11394737 TI - Protein ligand docking based on empirical method for binding affinity estimation. AB - An empirical protein-ligand binding affinity estimation method, SCORE, was incorporated into a popular docking program, DOCK4. The combined program, ScoreDock, was used to reconstruct the 200 protein-ligand complex structures and found to give good results for the complexes with high binding affinities. A quality assessment method for docking results from ScoreDock was developed based on the whole test set and tested by additionally selected complexes. The method significantly improves the docking accuracy and was shown to be reliable in docking quality assessment. As a docking tool in structural based drug design, ScoreDock can screen out final hits directly based on the predicted negative logarithms of dissociation equilibrium constants of protein-ligand complexes, and can explicitly deal with structure water molecules, as well as metal atoms. PMID- 11394738 TI - Design of new selective inhibitors of cyclooxygenase-2 by dynamic assembly of molecular building blocks. AB - A method of dynamically assembling molecular building blocks - DycoBlock - has been proposed and tested by Liu et al. This method is based on multiple-copy stochastic dynamics simulation in the presence of a receptor molecule. In this method, a novel algorithm was used to dynamically assemble the molecular building blocks to form candidate compounds. Currently, some new improvements have been incorporated into DycoBlock to make it more efficient. In the new version of DycoBlock, the binding energy and solvent accessible surface area (SASA) can be used to screen the resulting compounds. A simple clustering algorithm based on molecular similarity was developed and used to classify the remaining compounds. The revised DycoBlock was tested by breaking SC-558 - a selective inhibitor of cyclooxygenase-2 (COX-2) - into building blocks and reassembling them in the active site of the enzyme. The accuracy of recovery grew to 58.8% while it was only 16.7% in the previous version. Then, thirty-three kinds of molecular building blocks were used in the design of novel inhibitors and the investigation of diversity. As a result, a total of 1441 compounds was generated with high diversity. After the first screening procedure, there remained 864 reasonable compounds. The results from clustering indicate that the structural motifs in the diarylheterocycle class of COX-2-selective inhibitors have been generated using the revised DycoBlock, and their binding modes were investigated. PMID- 11394739 TI - Selective inhibition of 6-phosphogluconate dehydrogenase from Trypanosoma brucei. AB - A number of triphenylmethane derivatives have been screened against 6 phosphogluconate dehydrogenase from Trypanosoma brucei and sheep liver. Some of these compounds show good inhibition of the enzymes and also selectivity towards the parasite enzyme. Modelling was undertaken to dock the compounds into the active sites of both enzymes. Using a combination of DOCK 3.5 and FLEXIDOCK a correlation was obtained between docking score and both activity for the enzymes and selectivity. Visualisation of the docked structures of the inhibitors in the active sites of the enzymes yielded a possible explanation of the selectivity for the parasite enzyme. PMID- 11394740 TI - Classification of protein disulphide-bridge topologies. AB - The preferential occurrence of certain disulphide-bridge topologies in proteins has prompted us to design a method and a program, KNOT-MATCH, for their classification. The program has been applied to a database of proteins with less than 65% homology and more than two disulphide bridges. We have investigated whether there are topological preferences that can be used to group proteins and if these can be applied to gain insight into the structural or functional relationships among them. The classification has been performed by Density Search and Hierarchical Clustering Techniques, yielding thirteen main protein classes from the superimposition and clustering process. It is noteworthy that besides the disulphide bridges, regular secondary structures and loops frequently become correctly aligned. Although the lack of significant sequence similarity among some clustered proteins precludes the easy establishment of evolutionary relationships, the program permits us to find out important structural or functional residues upon the superimposition of two protein structures apparently unrelated. The derived classification can be very useful for finding relationships among proteins which would escape detection by current sequence or topology-based analytical algorithms. PMID- 11394742 TI - Molecular autopsy of sudden unexplained death in the young. AB - Sudden unexplained death (SUD) claims over 4,000 persons between the age of 1 and 22 each year in the United States. Nearly half of all pediatric SUD cases have a normal structural autopsy evaluation and are dismissed without a diagnosis. With the discovery of the genetic basis for potentially fatal arrhythmias associated with the inherited long QT syndrome (LQTS), postmortem molecular diagnosis of this disorder is possible. The authors describe the results of a molecular autopsy performed on a 17-year-old boy found dead in bed. A novel clinical test involving an epinephrine challenge in the decedent's mother implicated a potential defect in the phase 3 potassium current encoded by the gene KVLQT1. Exon-specific amplification by polymerase chain reaction and direct DNA sequencing of KVLQT1 revealed a 5-base pair deletion in the genetic material recovered from the decedent's paraffin-embedded heart tissue. The ability to perform molecular autopsies on archived necropsy material undoubtedly will transform the forensic evaluation of SUD. The combination of catecholamine provocation testing in survivors and a postmortem LQTS gene analysis may unmask families with "concealed" LQTS and establish the cause and manner of death in SUDS. PMID- 11394741 TI - Identification of cylin-dependent kinase 1 inhibitors of a new chemical type by structure-based design and database searching. AB - We have selected cyclin-dependent kinase 1 (CDK1), an enzyme participating in the regulation of the cell cycle, as a target in our efforts to discover new antitumor agents. By exploiting available structural information, we designed an ATP-site directed ligand scaffold that allowed us to identify 4-(3-methyl-1,4 dioxo-1,4-dihydro-naphthalen-2-ylamino)-benzenesulfonamide as a new potent inhibitor of CDK1 in a subsequent database search. The synthesis and testing of some analogues confirmed the interest of this class of compounds as novel CDK1 inhibitors. PMID- 11394743 TI - Position paper on fatal abusive head injuries in infants and young children. AB - This article represents the work of the National Association of Medical Examiners Ad Hoc Committee on shaken baby syndrome. Abusive head injuries include injuries caused by shaking as well as impact to the head, either by directly striking the head or by causing the head to strike another object or surface. Because of anatomic and developmental differences in the brain and skull of the young child, the mechanisms and types of injuries that affect the head differ from those that affect the older child or adult. The mechanism of injury produced by inflicted head injuries in these children is most often rotational movement of the brain within the cranial cavity. Rotational movement of the brain damages the nervous system by creating shearing forces, which cause diffuse axonal injury with disruption of axons and tearing of bridging veins, which causes subdural and subarachnoid hemorrhages, and is very commonly associated with retinal schisis and hemorrhages. Recognition of this mechanism of injury may be helpful in severe acute rotational brain injuries because it facilitates understanding of such clinical features as the decrease in the level of consciousness and respiratory distress seen in these injured children. The pathologic findings of subdural hemorrhage, subarachnoid hemorrhage, and retinal hemorrhages are offered as "markers" to assist in the recognition of the presence of shearing brain injury in young children. PMID- 11394744 TI - Survey of forensic pathologists. AB - A survey of board-certified forensic pathologists (from a list of board-certified forensic pathologists supplied by the American Board of Pathology) was conducted to determine their current practice situations. The purpose of the survey was to provide information useful to the public in general and to those organizations that represent forensic pathologists in particular to better understand and meet the needs of the forensic pathology community. A total of 773 surveys were mailed, and 18 were returned as undeliverable. There were 337 replies (45%). Responses were as follows: In terms of age, 7% were in their 30s; 32% were in their 40s; 29% were in their 50s; 19% were in their 60s; and 13% were 70 years of age or older. In terms of gender, 79% were male and 21% were female. For pathology practice status, 72% worked more than 30 hours per week; 11% between 5 and 30 hours per week; 5% less than 5 hours per week; and 12% no longer practice pathology. For percentage of pathology practice that is forensic pathology, 69% of respondents were more than 80%; 6% were 50%-80%; 5% were 25%-50%; 8% were 5% 25%; and 12% were less than 5%. In terms of forensic pathology practice setting, 54% were employed by medical examiner or coroner systems; 13% worked under contract to medical examiner or coroner systems; 27% worked in private practice or consultation; and 6% responded as "other." PMID- 11394745 TI - Asphyxial homicide in two Scandinavian capitals. AB - In the Oslo and Copenhagen capital areas, 94 asphyxial homicides were committed in the 10-year period 1985-1994, accounting for 22% of all homicides in that period. Sixty-nine (73%) of the asphyxia victims were female. The most common method of asphyxiation was manual strangulation. Seventeen (18%) of the victims were below the age of 10, accounting for 59% of all homicides in that age group. Whereas 38% of the female victims were killed by their spouse, this was the case for only one male victim. The motive was not known in a great proportion of cases. Fifty-seven percent of the victims had been subjected to additional violence, and in this respect there was no difference between the sexes. In 12 of the cases the offender was female; in 9 such cases the victim was her offspring. More than half of the victims had no blood alcohol. When disregarding the victims less than 10 years of age, 33% of the male and 49% of the female victims had no blood alcohol. The crime scene was the victim's domicile among 72% of female and 52% of male victims. Forty-two percent of the female and 11% of the male victims above the age of 10 years were married or cohabitant. PMID- 11394746 TI - Characteristic features of suicidal drownings: a 20-year study. AB - A retrospective study of cases of drowning suicide was undertaken at the Forensic Science Centre in Adelaide, South Australia for the period April 1980 to March 2000. A total of 123 cases were found, with 76 males (age, 16-88 years; average, 50.5 years; standard deviation [SD], 20.1 years) and 47 females (age, 34-88 years; average, 60.6 years; SD, 13.9 years). There were 66 fresh water drownings and 57 saltwater drownings. Female victims were significantly older than male victims for both fresh water and saltwater drownings (P < .05 and P < .01, respectively). Deaths in young women were rare. No temporal trend in drowning suicides could be shown, with annual numbers varying from 0 to 12 cases (average, 6.15; median, 7). Women preferentially chose the ocean or bath to drown themselves in, whereas males chose rivers, ditches, and lakes. Swimming pools were rarely used for suicide in this population; alcohol use was not usual; and there was often a significant history of mental illness. PMID- 11394747 TI - Morphologic determinants of asphyxia in lungs: a semiquantitative study in forensic autopsies. AB - Asphyxia is a name given to different kinds of lesions that can produce similar histologic findings. Thus, because of the varied nature of the different kinds of lesions, as well as the incidence of similar qualitative histologic findings with different causes, the aim of this work was to study special kinds of injuries with particular subsequent impairment. These include some diagnostic problems of sudden death of natural causes, including aspiration, suffocation, drowning, and strangulation. Ranking was made of 167 victims based on the diagnosis as having: aspiration (n = 35), suffocation (n = 88), drowning (n = 27), and strangulation (n = 17). Stepwise discriminant analysis of the resulting data showed that lung necropsies from victims of these four events could be distinguished from one another. Statistical differences among the four groups were observed for eight morphologic parameters. A robust discriminant function permitted an adequate classification of the four groups of disease in 85.03% of the cases. Lung autopsies with congestion, septal hemorrhage, and foreign body showed a specificity of 100% for victims of aspiration, whereas ductal overinsufflation, interstitial edema, and bronchiolar constriction showed a specificity of 81.8% in victims of suffocation. Intraalveolar edema and dilatation of the alveolar spaces with secondary compression of the septal capillaries characterized drowning. Victims of strangulation showed a strong alveolar hemorrhage, with alveolar collapse and overinsufflation, associated with bronchiolar dilatation. It is concluded that semiquantitative analysis of lung autopsies might be a useful supplementary histologic criterion to support the diagnosis of asphyxia. PMID- 11394748 TI - The forensic pathology of nonagenarians and centenarians: do they die of old age? (The Auckland experience). AB - The aim of this study is to provide awareness of the common causes of death and their associated trends in the very aged. Forensic autopsies on patients aged >90 years were reviewed. The study lasted from January 1, 1988 to December 11, 1998 and was done in Auckland, New Zealand, the population of which is 1.3 million. Cases were divided into natural or unnatural deaths. Of the total of 319 cases, 272 (85%) deaths were natural. Of those, only 13 (5%) were "written off" as being attributed to old age or senile debility. The most common causes of death were ischemic heart disease (IHD), 74 cases (23%); bronchopneumonia, 37 cases (12%); fractures, 28 cases (9%); acute myocardial infarction, 25 cases (8%); cerebrovascular accident, 19 cases (6%); and ruptured aneurysm, 17 cases (5%); 61 (19%) deaths were multifactorial. Fractures, either as the primary cause of death or as a complicating factor, accounted for 29 cases, third only to IHD and bronchopneumonia. Forty-seven deaths (15%) were unnatural; of those, 43 were accidents, 3 were suicides, and 1 was a homicide. From these results it is clear that the very elderly succumb to disease; they do not often die of old age. PMID- 11394749 TI - Death by overlaying and wedging: a 15-year retrospective study. AB - Overlaying, the accidental death by smothering caused by a larger individual sleeping on top of an infant, is a cause of death that has been documented for centuries. The hazard of death has been reported to be greater in infants less than 5 months of age but may occur in children up to the age of 2 years. When an adult or older child rolls on top of an infant, mechanical asphyxia results. The face may be pressed into the mattress or into the body of the sleeping adult or older child. The infant's air may be expressed, and he or she is unable to cry due to pressure on the thorax and the inability to inhale. Some pathologists and investigators believe that the victims of overlaying have no pertinent physical findings at autopsy and that any injury is indicative of inflicted trauma. Others believe that one may see contusions and abrasions from overlaying in and of itself. Wedging is another form of accidental mechanical asphyxia that may have negative autopsy findings. The prevalence of bruising, contusions, or facial and ocular petechiae is not clear. The author reviewed all pediatric forensic cases referred for autopsy to the Forensic Section of the Medical University of South Carolina/Medical Examiners' Office over the past 15 years, from 1985 to 1999. Of these, all cases of overlaying, cases listed as undetermined sudden infant death syndrome versus overlaying, and wedging were included. The cases were analyzed as to victims' age, sex, race, location/bedding, bed-sharer, and whether the bed sharer was known to have ingested drugs or alcohol before sleep. Postmortem physical findings were also reviewed, particularly for documentation of contusions, abrasions, or facial or ocular petechiae. By clarifying not only the victim, bed-sharer, and scenario but also the presence or absence of physical findings in cases of overlaying, wedging, and other accidental asphyxia, we can better categorize these cases. PMID- 11394750 TI - Heterotopic ossification in unidentified skeletal remains. AB - Heterotopic ossification is a benign, ectopic bone growth that develops in muscle and other soft tissue. The exact cause is poorly understood, but it is a rarely serious complication of soft tissue trauma. Its most common form, myositis ossifications traumatica, occurs as a secondary complication of direct muscle injury. However, other forms are less common and can result from specific pathologic conditions, such as spinal cord trauma and metabolic disorders. In patients who have had spinal cord injury and subsequent paraplegia, heterotopic ossification often results in ankylosis of the hip and a loss in range of motion. Ectopic ossification occurs below the injury site, and, although the specific muscle groups can vary, it usually involves those for which the origin and insertion involve the anterior pelvis and proximal femur. In dried bone, heterotopic ossification can appear as a smooth, irregularly shaped benign tumor of mature bone, extending from the surface but not invading the cortical bone. These tumors range in size from a few millimeters to several centimeters. Because heterotopic ossification is often associated with specific types of injuries, it has a unique anthropological use in forensic cases. PMID- 11394751 TI - Echinococcal hydatid cyst at the right ventricle outlet with leakage to the pulmonary artery outflow causing follicular airway disease and sudden death. AB - Postmortem examination of a 35-year-old parous woman who died suddenly revealed a hydatid cyst mass located at the right ventricular outlet, with a grossly discernible defect opening to the pulmonary outflow tract. Pulmonary hemorrhage, follicular bronchitis, and bronchiolitis also were present, with severe acute purulent exudation in the airways. Hydatid cyst complications must be kept in mind when dealing with sudden deterioration and death in patients who are residents of regions where echinococcosis is endemic. PMID- 11394752 TI - Elderly suicide: a 10-year retrospective study. AB - The suicide rate in the 65-year and older age group has been increasing since 1980. The elderly attempt suicide less often than younger people but are successful more often. The authors retrospectively reviewed all cases referred to the Forensic Pathology Section of the Medical Examiners' Office at the Medical University of South Carolina (Charleston, SC) from January 1988 through December 1997. The cases of suicide in victims 65 years and older totaled 78, accounting for 11.5% of all suicides reviewed. Of these 78 cases, 41% were autopsied and 59% were externally examined. All of the cases were initially analyzed as to age, race, sex, and method of suicide. Files also were reviewed to determine time of year of the suicide, toxicology results, psychiatric history, social history, medical history, and whether a suicide note was left. The ages of these suicide victims ranged from 65 to 94 years; men comprised 85% of the victims, and whites, 94%. The male-to-female and white-to-black ratios were 6:1 and 15: 1, respectively. The average age of the victim was 73 years. Gunshot wound was the most common method of suicide, accounting for 80.7% of the cases. Other methods included overdose (6.4%), hanging (3.8%), fall from height (2.6%), incised wounds (2.6%), drowning (2.6%), and carbon monoxide poisoning (1.3%). PMID- 11394753 TI - A rapid postmortem cardiac troponin T assay: laboratory evidence of sudden cardiac death. AB - Postmortem examination may be useful in establishing the cause of sudden unexpected death. In many instances, however, limitations of staffing, budget, and time may force the pathologist to triage cases to external examination rather than autopsy. A rapid assay for cardiac troponin T (cTnT) to document suspected cardiac-related deaths may optimize the use of the time and resources of the autopsy pathologist. Peripheral blood was sampled percutaneously before each of 40 autopsies and placed in the well of the Cardiac T Rapid Assay unit in accordance with the included instructions, and the results were read after 15 minutes. The assay result, decedent age, postmortem interval, and evidence of cardiopulmonary resuscitation were tabulated and subsequently correlated with the cause of death. On final sign-out of each of the autopsies, the cause of death was determined to be cardiac-related (n = 20) versus the cause in non-cardiac control subjects (n = 20). This determination was made while the investigators were blinded to the cTnT assay result. Of the 20 cardiac deaths, 17 (85%) showed positive results for cTnT compared with 6 (30%) false-positive results among the 20 control cases; this result was statistically significant according to the chi square test. In the over-50 age group, the sensitivity of this assay in detecting cardiac-related death was 91%, with a specificity of 86%. Perimortem cardiopulmonary resuscitation did not appear to result in false-positive results. In the appropriate setting, this rapid assay for cTnT can provide valuable data supportive of a cardiac-related death. This inexpensive test may best be used in triaging sudden deaths in persons over 50 to external examination versus complete autopsy. PMID- 11394754 TI - Acute necrotizing bacterial tonsillitis with Clostridium perfringens. AB - Bacterial infection with Clostridium perfringens in children less than 2 years of age is frequently associated with meningitis, necrotizing gastrointestinal infection, and postoperative infections. However, a review of the literature reveals no reports of these bacteria infecting the tonsils. A 9-month old black female was found unresponsive at the baby-sitter's and was rushed to the hospital. Shortly after admission to the emergency department death was pronounced. An autopsy performed on this otherwise healthy infant revealed shock and acute necrotizing bacterial tonsillitis. The initial report of this infant's death was questionable sudden infant death syndrome and questionable smothering. Postmortem cerebrospinal fluid, blood, and lung cultures grew pure colonies of C. perfringens. The necrotizing tonsil revealed no significant gross lesions. Microscopically, large numbers of gram-positive rods were easily recognized and were compatible with C. perfringens. Because the oropharynx is a common portal of entry for infectious agents, it is essential to sample tissues of Waldeyer's ring and especially the tonsils to find infectious diseases that may become systemic. PMID- 11394755 TI - Complete autoerotic asphyxiation: suicide or accident? AB - A case of a typical form of sexual asphyxiation is described, involving a young man hanging, in a context of autoerotic asphyxia. Multiple and different parameters are included in the definition. PMID- 11394756 TI - Pediatric toxicologic deaths: a 10-year retrospective study. AB - A 10-year retrospective study of pediatric toxicologic deaths was performed at the Medical University of South Carolina (Charleston, SC) from January 1989 to December 1998. During this time, 709 pediatric forensic autopsies were performed on children younger than 18 years of age. Eleven deaths were determined to be secondary to toxic exposures (excluding carbon monoxide poisonings secondary to fires). The remaining deaths were reviewed for the presence of alcohol or illicit drugs. The 11 toxicologic deaths were analyzed for age, sex, race, type of toxic exposure, cause and manner of death, location of incident, witness, and, in the younger age group, the primary caregiver at the time of exposure. The deaths had a bimodal age distribution (6 deaths in victims ages 15 to 17 and 5 deaths in victims ages 4 or younger), involving a wide range of toxins. The teenage group was composed of five males and one female, all white. The preschool group had three females and three males, all black. The manner of death ranged from accidental to suicidal to homicidal. In addition, in eight neonatal and fetal deaths, the victims tested positive for maternal cocaine use, and five of these victims tested positive for cocaine or benzoylecgonine. However, the cause of death was not stated to be cocaine in any of these neonatal and fetal cases. PMID- 11394757 TI - Pulmonary hemorrhage in deceased infants: baseline data for further study of infant mortality. AB - Infant lung samples were obtained at autopsy by medical examiner pathologists in five areas of the United States regardless of the cause of death. Lung sections were stained with hematoxylin and eosin. Sixty cases were evaluated for the study. The four sections examined for each case were taken from the anterior and posterior aspects of the right and left upper lung lobes. Histologic sections were scored for the extent of alveolar hemorrhage using the following scoring system: 0, no hemorrhage; 1, focal hemorrhage but less than score 2; 2, patchy, focal hemorrhage not present throughout the section; 3, focal hemorrhage more extensive than score 2 but not meeting the criteria for score 4; 4, patchy focal hemorrhage distributed throughout the section; 5, more extensive hemorrhage than score 4 but not meeting the criteria for score 6; 6, diffuse hemorrhage throughout the section. Total possible scores ranged from 0 to 24. Intraalveolar hemorrhage was observed in 40 cases. Overall, the mean score for the 60 cases was 5 (range, 0-24); for the 40 cases with hemorrhage, 7 (range, 1-24). Scores were compared with other descriptive variables like cause of death; interval between onset of fatal events and death; whether resuscitation was attempted; and pulmonary macrophage counts and hemosiderin scores reported in earlier studies of the same cases. In none of the 60 cases was death attributed to pulmonary hemorrhage or hemosiderosis. Pulmonary hemorrhage tends to be common among deceased infants; more prominent when there is medical treatment or resuscitation during the agonal period; infant position may partially explain distribution of hemorrhage in lungs; postmortem interval may exacerbate pulmonary hemorrhage; and infant deaths caused by acute idiopathic pulmonary hemorrhage (AIPH) or pulmonary hemorrhage/hemosiderosis (PHH) probably are rare. Specific case definitions for AIPH and PHH are needed, along with further study of these conditions. PMID- 11394758 TI - Asphyxiation from shoulder seat belts: an unusual motor vehicle injury. AB - Two cases of accidental asphyxia caused by lap-shoulder seat belts are reported. Lethal neck injuries from shoulder seat belts are uncommon, and episodes of asphyxiation are even rarer. Positioning of the shoulder belt over the neck, short stature, and incapacitation from other injuries were predisposing factors to upper airway compromise in these cases. PMID- 11394759 TI - An analysis of factors contributing to a series of deaths caused by exposure to high environmental temperatures. AB - Autopsy reports at the Forensic Science Centre, Adelaide, South Australia, were reviewed for the 8 years from January 1991 to December 1998 for cases with unusual features in which deaths had been attributed to exposure to high environmental temperatures. Amphetamine-related hyperpyrexial deaths, anesthetic deaths caused by malignant hyperpyrexia, deaths of elderly incapacitated individuals during heat waves, and deaths of children trapped in the back of cars were excluded from the study. In 9 cases, where heat-related deaths had occurred (age range 21 to 77 years; M:F = 8:1). Predisposing factors included lack of familiarity with Australian environmental conditions, excessive clothing, prolonged sun exposure, acute alcohol intoxication, obesity, benztropine and trifluoperazine medication, and underlying dementia, alcoholic liver disease, and possibly epilepsy. PMID- 11394760 TI - Frequency of bronchopneumonia in children with survival interval before death. AB - Many children do not survive after presentation in extremis. Some survive varying intervals and are found to have bronchopneumonia at death. The question is raised whether bronchopneumonia is a consequence of survival rather than the initiating disease leading to collapse. A prospective study of the deaths of 156 children divided them into two groups: 80 children with head injury and 76 with causes of death other than sudden infant death syndrome. In 43 of the total group of children, bronchopneumonia was found. In the total group, 76 survived more than a day. Of these 39 had bronchopneumonia, 32 died of head injury, and 7 had other causes of death. Of the children surviving less than a day, 4 had bronchopneumonia at death--only 1 with head injury. If bronchopneumonia is present, it is more likely to have developed after the collapse than to have caused it in this population. PMID- 11394761 TI - Colchicine poisoning: case report of two homicides. AB - Deaths resulting from the oral ingestion of colchicine are occasionally associated with suicides and therapeutic toxicity. However, homicidal deaths in which this drug is used are extremely rare. Two homicides in which colchicine was used are reported. PMID- 11394762 TI - Unexpected childhood death due to a rare complication of ventriculoperitoneal shunting. AB - A 10-year-old boy with Arnold-Chiari malformation, spina bifida, and a ventriculoperitoneal shunt for hydrocephalus died unexpectedly, having appeared to be only mildly unwell with fever on the night before death. At autopsy, the shunt was partially obstructed with an associated enterococcal meningitis. The tip of the shunt was located within the transverse colon, which was embedded in a mass of fibrous adhesions resulting from previous abdominal surgery. Blood cultures were sterile. Intestinal perforation is a rare complication of ventriculoperitoneal shunting that may be associated with the development of meningitis and unexpected death. The autopsy assessment of children with such indwelling devices requires examination of the functional state of the shunt, full septic workup, and determination of the precise location of the tip of the catheter within the peritoneal cavity. PMID- 11394763 TI - SIDS doesn't exist. PMID- 11394764 TI - Suicidal incised wound of a fistula for hemodialysis access in an elderly woman. PMID- 11394765 TI - Relationship between arousal reaction and autonomic nervous system in the sudden infant death syndrome. PMID- 11394766 TI - Rapid removal of dilute lead from water by pyroaurite-like compound. AB - Rapid removal of dissolved dilute lead (Pb) by pyroaurite-like compound Mg(x)Fe(y)(OH)2(x+y) (CO3(2-))y/2 x mH2O; anionic clay: one of layered double hydroxides) from water was studied through batch and column experiments. The Pb removal property of the compound was evaluated as a function of concentration of Pb in the treated solution, space velocity (Sv) of the solution in the column packed with the compound, pH of the solution and contaminated humic substance. The compound showed a highly effective Pb-removal property that was comparable to the other conventional adsorbents. The Pb was removed rapidly by the column packed with the compound over the wide range of the Sv. The buffering pH function of the compound contributed to the Pb removal producing weak-alkali atmosphere. The contamination of humic substance in the treated solution reduced the Pb removal largely depending on its amount. The effectiveness of the compound for the rapid Pb removal was confirmed through a series of experiments. PMID- 11394767 TI - Survival and precopulatory behaviour of Gammarus pulex (L.) exposed to two xenoestrogens. AB - The effects of the xenoestrogens ethinylestradiol (EE) and bisphenol A (BPA) on the survival and reproductive behaviour of the amphipod Gammarus pulex were determined in a series of bioassays. Acute toxicity of the test chemicals was determined under static conditions, revealing 10 d median lethal concentrations (LC50s) of 0.84 and 1.49 mg/L for EE and BPA, respectively. Several aspects of the reproductive behaviour including the ability of males and females to detect each other, form precopulatory guarding pairs and to continue the guarding behaviour, were examined during a 24 h exposure period over a wide range of concentrations. However, reproductive behaviour was only disrupted at relatively high concentrations where it would be unrealistic to attribute the effects to an endocrine mediated process. Consequently, changes in precopulatory guarding resulting from acute exposure. do not seem to be suitable for detecting the presence in water of xenoestrogens known to cause endocrine disruption in vertebrates. PMID- 11394768 TI - Characteristics of nitrogen and phosphorus loads in peat mining wastewater. AB - Runoff water quality from a peat mine was studied during 1995 and 1996 in Central Finland. Water samples from three drained sub-catchments and groundwater were analysed for all the standard physio-chemical parametres including different forms of nitrogen and phosphorus. The annual leaching of phosphorus, nitrogen and suspended solids was estimated to be 16-38 kgkm(-2), 1,073-1,500 kgkm(-2), and 2 8 tkm(-2), respectively. The variation in nutrient concentrations could be best explained by the relative volumes of new water (5-day sum of rainfall), ditchwater temperature and conductivity. Heavy rainfall caused new water to infiltrate washing nitrate out of the unsaturated peat layer resulting in high concentrations in ground- and ditchwater. Ditchwater phosphorus concentrations always decreased with increased runoff and peaked, as did COD and colour, after dry spells when old groundwater dominated runoff. A large part of the suspended solids load occurred during snowmelt, whereas dissolved solids and nitrogen loads peaked during summer flows. PMID- 11394769 TI - Effectiveness of an anaerobic granular activated carbon fluidized-bed bioreactor to treat soil wash fluids: a proposed strategy for remediating PCP/PAH contaminated soils. AB - An integrated system has been developed to remediate soils contaminated with pentachlorophenol (PCP) and polycyclic aromatic hydrocarbons (PAHs). This system involves the coupling of two treatment technologies, soil-solvent washing and anaerobic biotreatment of the extract. Specifically, this study evaluated the effectiveness of a granular activated carbon (GAC) fluidized-bed reactor to treat a synthetic-waste stream of PCP and four PAHs (naphthalene, acenaphthene, pyrene, and benzo(b)fluoranthene) under anaerobic conditions. This waste stream was intended to simulate the wash fluids from a soil washing process treating soils from a wood-preserving site. The reactor achieved a removal efficiency of greater than 99.8% for PCP with conversion to its dechlorination intermediates averaging 46.5%. Effluent, carbon extraction, and isotherm data also indicate that naphthalene and acenaphthene were removed from the liquid phase with efficiencies of 86 and 93%, respectively. Effluent levels of pyrene and benzo(b)fluoranthene were extremely low due to the high-adsorptive capacity of GAC for these compounds. Experimental evidence does not suggest that the latter two compounds were biochemically transformed within the reactor. PMID- 11394770 TI - Water quality of the odzi river in the Eastern Highlands of Zimbabwe. AB - For Odzi. the main river originating in the Eastern Highlands of Zimbabwe, the nature, sources and extent of pollution are characterised. Over a span of 9 months, the river water samples collected at six selected sites were analysed for various physical and chemical parameters namely, temperature, conductance, pH, total suspended and dissolved solids, BOD. total phosphate and nitrate levels. The Mutare river carried the seepage from abandoned mine dumps to the Odzi River. Water quality of the Odzi River at different points assessed by the water quality indices (WQI) is compared. According to the quality indices during the study period, water quality in the upper reaches of the river was medium to good. It dwindled in the plains, due to the seepage from abandoned mine dumps and discharges from the farm lands. PMID- 11394771 TI - Experimental analysis of centrifugal dewatering process of polyelectrolyte flocculated waste activated sludge. AB - The study experimentally investigated the centrifugal separation of moisture from activated sludge subject to cationic polyelectrolyte flocculation. An arm suspended centrifuge was employed which allowed in-situ detection on all positions of interfaces of centrifuged sludge as functions of time. Experimental results revealed that, sludge flocculation would yield a significant sedimentation effect at the first phase of centrifugation. Therefore, not as suggested in conventional centrifugal-filtration models, the most significant moisture-removal stages included filtrate to flow through a wet cake. Moreover, an optimal rotational speed exists at which the moisture-removal rate reaches a maximum value. New theories/correlations are required to describe centrifugation dewatering of polyelectrolyte flocculated sludge. PMID- 11394772 TI - Equilibrium and kinetics of metal ion adsorption onto a commercial H-type granular activated carbon: experimental and modeling studies. AB - Systematic studies on metal ion adsorption equilibrium and kinetics by a commercial H-type granular activated carbon were carried out. Titration of the carbon showed that the surface charge density decreased with an increasing pH. Higher copper adsorption was obtained with increasing solution pH and ionic strength. Metal removal was in the descending order: Cu2+ > Zn2+ approximately Co2+. Copper removal was not affected by addition of zinc or cobalt, while copper can reduce both zinc and cobalt removal. Kinetic experiments demonstrated that the copper adsorption rapidly occurred in the first 30-60 min and reached the complete removal in 3-5 h. Removal of zinc and cobalt was slightly slower than that of copper. It was found that the mass transfer is important in the metal adsorption rate. The surface complex formation model was used successfully to describe the surface change density, as well as the single- and multi-species metal adsorption equilibrium. The copper removal was due to adsorption of Cu2+, CuOH+, and CuCl+, while the zinc and cobalt uptake was due to the formation of surface metal complexes of SOM2+ and SOMOH+ (M = Zn and Co). It was found that the diffusion-control model well described the adsorption kinetics with various metal ions and pH values. Finally sensitivity analysis on the kinetic model's parameters was carried out. PMID- 11394773 TI - Effect of functional groups of humic substances on uf performance. AB - The role of different functional groups present in humic substances on the membrane flux is unclear. This study is undertaken to (1) separate the carboxyl and phenolic groups from a humic solution, and (2) evaluate the effect of each fractionated humic substances on the ultrafiltration (UF) performance. A weak base amine resin was used for the adsorption (pH 7) and the subsequent desorption (pH 13) of the phenolic groups from a commercial humic solution. These fractions were evaluated qualitatively (via Fourier transform infrared spectroscopy) and quantitatively (titration); they were further subjected to the analyses of the trihalomethane formation potential (THMFP) and ultrafiltration performance. Although, a complete separation of the phenolic and carboxyl groups is not possible, the results nevertheless provide useful information about their effects on UF performance. The fraction with a higher content of the phenolic OH group exhibits the highest THMFP (190 microg/mg C), whereas the fraction with a higher content of the carboxyl groups exhibits more flux decline. The UF system evaluated is unable to remove a significant portion of THM precursors, resulting in high THMs in permeate. The use of powdered activated carbon for the pretreatment of these fractions fails to improve membrane fouling. The pore size of UF membrane does not appear to affect the membrane flux, and the switch from the hydrophobic to hydrophilic membrane only slight improves the permeate flux. PMID- 11394774 TI - Simultaneous organic and nitrogen removal from municipal landfill leachate using an anaerobic-aerobic system. AB - An anaerobic-aerobic system including simultaneous methanogenesis and denitrification was introduced to treat organic and nitrogen compounds in immature leachate from a landfill site. Denitrification and methanogenesis were successfully carried out in the anaerobic reactor while the organic removal and nitrification of NH4+,-N were carried out in the aerobic reactor when rich organic substrate was supplied with appropriate hydraulic retention time. The maximum organic removal rate was 15.2 kg COD/m3 d in the anaerobic reactor while the maximum NH4+-N removal rate and maximum nitrification rate were 0.84kg NH4+ N/m3/d and 0.50kg NO3--N/m3/d, respectively, in the aerobic reactor. The pH range for proper nitrification was 6-8.8 in the aerobic reactor. The organic compounds inhibited nitrification so that the organic removal in the anaerobic reactor could enhance the nitrification rate in the following aerobic reactor. The gas production rate was 0.33 m3/kg COD and the biogas compositions of CH4, CO2, and N2 were kept relatively constant, 66-75, 22-32, and 2-3%, respectively. PMID- 11394775 TI - Toxicity characterisation of organic contaminants in stormwaters from an agricultural headwater stream in south east England. AB - The transient movement of pesticides at biologically active concentrations during storm events is considered to be a cause of biological impoverishment in some headwater streams. The programme of work described sought to identify compounds that are the cause of toxic effects during such events. Along with targeted pesticide analysis, toxicity identification evaluation (TIE) procedures were used to identify compounds with a demonstrated toxic effect. These procedures were specifically directed towards isolating and attributing toxicity to classes of organic contaminants in samples collected from an English headwater stream during a storm event. The organic load was isolated by means of solid-phase extraction (SPE). Bioassay of the SPE extract at x 100 whole water concentrations confirmed that the samples contained substances toxic to Daphnia magna, although the raw samples were not toxic. Targeted pesticide analysis identified simazine and diuron as the major pesticides present and, using a toxicity unit (TU) approach. were shown to be responsible for a significant amount of the observed concentrate toxicity during a runoff event. However, they were not present in sufficient quantities to be totally responsible for a more toxic later event. By simplification of the SPE isolate using reverse-phase HPLC, fractions from which were tested for toxicity, the cause of concentrate toxicity in the later event was isolated to two discrete fractions. GC-MS analysis of these fractions identified nonylphenol. endosulfan sulphate and pendimethalin as present, with the majority of toxicity attributed to nonylphenol (NP). The main advantage of the TIE approach is that it allows biological active compounds with a demonstrated effect to be identified that may not be selected by more traditional techniques. PMID- 11394777 TI - Aggregation rates of natural particle populations. AB - In this paper an experimental approach of aggregation in natural suspensions is presented. The suspensions are organic-matter-rich waters sampled in a brook which drains peat areas. The aggregation was conducted on raw samples in three different experiments lasting from 2 to 8 days. The particle size distribution (PSD) in the 0.5-10 microm size range was followed with a laser sizer and appeared to be almost constant along the whole experiment duration. Nevertheless, the volume of particles larger than 10 microm increased steadily, showing that aggregation occured. This appeared to be the consequence of a steady-state aggregation which allowed the removal of the whole particle set within a day. The use of an aggregation model adapted to calculations on PSD allowed estimation of the aggregation efficiency for such suspensions. PMID- 11394776 TI - Correlation analyses on binding behavior of heavy metals with sediment matrices. AB - This article presents the amounts of heavy metals bound to the sediment matrices (carbonates, Fe-oxides, Mn-oxides, and organic matter), the correlations between any two heavy-metal binding fractions, and the correlations between sediment matrices and their heavy-metal binding fractions. Data consisted of 313 sets obtained from five main rivers (located in southern Taiwan) were analyzed by statistical methods. Among six heavy metals analyzed (Zn, Cu. Pb, Ni, Cr, and Co), the statistical results show that Zn is primarily bound to organic matter, and Cr is primarily bound to Fe-oxides. Principal component analysis (PCA) and correlation analysis (CA) result in significant correlations between carbonates bound Ni and carbonates bound Cr, Fe-oxides bound Ni and Fe-oxides bound Cr, and Mn-oxides bound Cu and Mn-oxides bound Cr. From linear regression results, the levels of the six heavy metals bound to either organic matter or Fe-oxides is moderately dependent on the contents of organic matter or Fe-oxides, especially true for Cr and Pb. According to slope values of linear regression, Cu and Cr have the highest specific binding amounts (SBA) to organic matter and Fe-oxides, respectively. A significant correlation between organic matter and organically bound heavy metals implied that organic matter contained in the sediments of the Potzu river and the Yenshui river can be adequately used as a normalizing agent. However, the six heavy metals bound to either carbonates or Mn-oxides do not correlate with carbonates or Mn-oxides. The obtained results also imply that competitions of various sediment phases in association with heavy metals occur, and organic matter and Fe-oxides are more accessible to heavy metals than other sediment phases. PMID- 11394778 TI - Comparison of the filtration characteristics between attached and suspended growth microorganisms in submerged membrane bioreactor. AB - An attached growth bioreactor was designed to minimize the effect of suspended microorganisms on membrane fouling in submerged membrane bioreactor. Comparison of mixed liquor from attached and suspended growth systems was made to elucidate major factors giving rise to different filtration characteristics. Unexpectedly, the rate of membrane fouling of the attached growth system was about 7 times higher than that of the suspended growth system despite similar characteristics of soluble fraction from the two reactors. Filtration performance proved to depend on the concentration of mixed liquor suspended solids (MLSS). Better filtration performance with suspended growth was explained by the formation of dynamic membranes with suspended solids. A series of analyses such as hydraulic resistance, specific cake resistance, scanning electron microscope, and atomic force microscope were carried out to elucidate the different filtration characteristics of the two systems. PMID- 11394779 TI - Effect of temperature increase from 55 to 65 degrees C on performance and microbial population dynamics of an anaerobic reactor treating cattle manure. AB - The effect of a temperature increase from 55 to 65 degrees C on process performance and microbial population dynamics were investigated in thermophilic, lab-scale, continuously stirred tank reactors. The reactors had a working volume of 31 and were fed with cattle manure at an organic loading rate of 3 g VS/l reactor volume/d. The hydraulic retention time in the reactors was 15 days. A stable reactor performance was obtained for periods of three retention times both at 55 degrees C and 65 degrees C. At 65 degrees C methane yield stabilized at approximately 165ml/g VS/d compared to 200 ml/g VS/d at 55 degrees C. Simultaneously, the level of total volatile fatty acids, VFA, increased from being below 0.3 g/l to 1.8-2.4 g acetate/l. The specific methanogenic activities (SMA) of biomass from the reactors were measured with acetate, propionate, butyrate, hydrogen, formate and glucose. At 65 degrees C. a decreased activity was found for glucose-, acetate-, butyrate- and formate-utilizers and no significant activity was measured with propionate. Only the hydrogen-consuming methanogens showed an enhanced activity at 65 degrees C. Numbers of cultivable methanogens, estimated by the most probable number (MPN) method, were significantly lower on glucose, acetate and butyrate at the increased operational temperature, while the numbers of hydrogenotrophic methanogens remained unchanged. No viable propionate-degrading bacteria were enriched at 65 C. Use of ribosomal oligonucleotide probes showed that an increase in temperature resulted in a decreased contribution of the rRNA of the domain bacteria from 74-79 to 57 62% of the universal probe, while the rRNA of the domain archaea, increased from 18-23 to 34-36%. PMID- 11394780 TI - Photodegradation mechanism and rate improvement of chlorinated aromatic dye in non-ionic surfactant solutions. AB - A typical insoluble chlorinated aromatic dye (CAD), disperse red (DR), was used to explore the reaction mechanism and kinetics of photodegradation in non-ionic surfactant solutions. The use of an additional hydrogen source and photosensitizer is also studied to improve the decay rates. The decay rate of dye in surfactants depends on the Km of surfactants and their ability to offering an effective hydrogen source. The photodegradation of CAD can be divided into three stages: the initial lag stage. the fast degradation stage and the final retardation stage. The lag stage will vanish and the decay rates of dye can be greatly improved by 2.5-3.6 times after adding an additional hydrogen source (NaBH4) or photosensitizer (acetone) to the surfactant micellar solution. However, the use of an additional hydrogen source or photosensitizer has dosage limitations in such applications. The photoreduction of DR is the main reaction mechanism, in which photodechlorination is observed first with the generation of HCI as the final product, then followed by photodecolorization by breaking the azo bond of the chromophore. PMID- 11394781 TI - Simplified analysis of contaminant rejection during ground- and surface water nanofiltration under the information collection rule. AB - A simple, closed-form analytical expression based on the homogenous solution diffusion model is derived for contaminant removal during nanofiltration (NF) of ground and surface water. Solute permeation and back-diffusion coefficients were used as fitting parameters to model rejection characteristics of four thin-film composite NF membranes under conditions typical of drinking water NF. Nonlinear fits of the model to experimental data suggests that the United States Environmental Protection Agency's (USEPA)'s Information Collection Rule protocol for bench-scale studies could be improved to obtain greater precision of the mass transfer coefficients. The model was found to fit rejection data for several water treatment contaminants including total organic carbon, precursors to total organic halide, four trihalomethanes and nine haloacetic acids containing chlorine and bromine, calcium and total hardness, alkalinity and conductivity. The simplified approach to mass transfer calculations from multisolute systems suggests that feed water recovery has a stronger influence on contaminant rejection than permeate flux. Evidence for coupled transport of divalent inorganic ions is also presented. Even though the model developed does not account for ion coupling and cannot be applied in a purely predictive mode, it can assist in the better design and interpretation of data obtained from site specific pilot-scale water treatment NF studies conducted in support of plant design. PMID- 11394782 TI - The electronic nose as a rapid sensor for volatile compounds in treated domestic wastewater. AB - An electronic nose consisting of 12 metal oxide sensors was used to monitor volatile compounds in effluent of a domestic wastewater treatment plant. Effluent and reference (deionized water) samples were heated to 60 and 90 degrees C to promote the volatilization and to increase the sensitivity. An effluent measuring campaign of 12 weeks was conducted and the repeatability and reproducibility of the procedure and the apparatus were determined. Processing the obtained fingerprints with principal component analysis (PCA) allowed interpretation and differentiation of the samples in terms of origin and quality, relative to the reference. To minimize the variance due to sensitivity fluctuations of the apparatus and to detect effluents with deviating qualities, two new concepts were defined, i.e. the relative sensorial odour perception (in short: rSOP) and the relative fingerprint. Correlations between the relative overall electronic nose output, expressed as rSOP, and selected routine parameters were weak except for the parameter "volatile suspended solids" (VSS), indicating adsorption of volatile organic compounds (VOCs) onto the organic particles. The results clearly demonstrate the possibility to use the electronic nose as a rapid alarm generator towards volatile compounds, e.g. in specific advanced treatment processes to produce reclaimed water from effluent of the domestic wastewater treatment plant under scrutiny. PMID- 11394783 TI - Extracellular enzyme activities during slow sand filtration in a water recharge plant. AB - Activities of the extracellular enzymes beta-glucosidase and phosphatase and bacterial densities were investigated during the filtration process at several sites in a groundwater recharge plant at the Ruhr river (Hengsen recharge plant in Schwerte. Germany). Low numbers of microorganisms and low levels of activity in this type of habitat, compared to most surface waters, caused methodological problems when determining microbial activity. In this study, fluorigenic model substrates, which enable hydrolytic rates as low as 1 nmol (L x h)(-1) to be measured, were used to determine extracellular enzyme activities. Highest activities were determined in surface water (107 nmol (L x h)(-1) for beta glucosidase and 252 nmol (L x h)(-1) for phosphatase). which decreased during the filtration process in the gravel prefilter and the main sand filter until the end of subsurface flow (1.6 nmol (L x h)(-1) and 6.8 nmol (L x h)(-1), respectively). Similarly, bacterial numbers decreased from 3.4 x 10(6) to 0.29 x 10(6) cells mL( 1). These data showed that microbial activity within the prefilter and the shallow layers of the sand filter had the greatest impact on water quality. In addition to its involvement in the continuous purification of surface water, the microbial community in the sand filter probably acts as a biological buffer against ephemeral increases in the loads of organic matter and nutrients in the recharge plant. PMID- 11394784 TI - Physicochemical speciation of molybdenum in rain water. AB - A combination of a sensitive catalytic determination method with filtration and ultrafiltration has been used for the physicochemical speciation of molybdenum in natural and synthetic rain water samples. The concentration (CL) of labile molybdenum was evaluated by a direct catalytic determination. The total concentration (CT ) of molybdenum was determined after the acid decomposition of the sample to calculate the non-labile concentration (CT-CL). From the speciation results, molybdenum species in the successive rainfall sample were found in a fraction with smaller molecular weights < 10(3) Da and characterized as labile forms, i.e. simple molybdate ions. Non-labile molybdenum existed in particulate matter (> or = 0.45 microm in particle size) and distributed predominantly in the initial rainfall sample. The coprecipitation with Fe(III) hydroxide contributed to the formation of the non-labile molybdenum. In the initial rainfall sample, a small part of molybdenum was labile in the particle fraction (> or = 0.45 microm). This type of molybdenum was associated with the formation of humic iron aggregates. PMID- 11394785 TI - In situ polychlorophenol bioremediation potential of the indigenous bacterial community of boreal groundwater. AB - The composition and chlorophenol-degrading potential of groundwater bacterial community in a permanently cold, oxygen-deficient chlorophenol contaminated aquifer at Karkola, Finland was studied with the aim of evaluating in situ bioremediation potential. The groundwater contained from 10(4) to 10(7) microscopically counted cells/ml and up to 10(5) CFU/ml heterotrophic bacteria cultivable at 8 and 20 degrees C. Of the 102 pure cultures, of which 86% Gram negative, from the plume area (10,000 microg of chlorophenols/l), 57% degraded 2, 3, 4, 6-tetrachlorophenol (TeCP), the main component of the wood preservative which was the source of contamination: 17% also degraded pentachlorophenol (PCP). The degraders were scattered among 16 different clusters of Gram-negatives mainly proteobacteria and members of Cytophaga/Flexibacter/Bacteroides phylum judged by the composition of whole-cell fatty acids. Only one Gram-positive degrading cluster was found containing seven actinobacteria closest to Nocardioides. Of the 88 pure cultures isolated from outside the plume (< 10 microg of chlorophenols/l) 67% were Gram-negative. Seven percent of the isolates degraded 2, 3. 4, 6-TeCP and/or PCP. Five of the Gram-positive isolates from outside the plume were Mycobacterium/Rhodococcus-related actinobacteria and O-methylated 2, 3, 4, 6-TeCP and PCP. The results show that chlorophenol degrading bacterial flora had been enriched as a result of contamination of the aquifer. This suggests significant in situ bioremediation potential of the site. PMID- 11394786 TI - Measurement of biodegradable dissolved organic carbon (BDOC) in lake water with a bioreactor. AB - Once-through, plug-flow bioreactors were colonised and maintained with a microbial community from a mesotrophic lake and used to measure the concentration of biodegradable dissolved organic carbon (BDOC). A BDOC measurement can be done within 3-4h by this method. Glucose was used to test whether oxygen consumption (BOD) could substitute for measurements of dissolved organic carbon (DOC). All added glucose was utilised, however, without a concomitant increase in oxygen demand. Oxygen consumption should not be used in bioreactor measurements. The site-specificity was tested by comparing DOC utilisation in bioreactors with batch cultures inoculated with indigenous bacteria and incubated for 28 days. The bioreactors were not site-specific and required no acclimation to measure BDOC from three different systems. However, humic substances were adsorbed in the reactors and about two days were needed to equilibrate the reactors. The BDOC concentrations in two lakes varied 2-fold over diurnal cycles and 3-fold during the period February-June. No significant relations to the light, dark cycle, chlorophyll, and DOC were found. The absolute BDOC concentrations ranged from 20 to almost 200 microM and averaged 13% of the DOC in the lakes. It is concluded that BDOC in lakes and other fresh waters can be measured quickly and reliably with a bioreactor. PMID- 11394787 TI - The effects of media size on the performance of biological aerated filters. AB - Biological aerated filters (BAFs) are an attractive process option, particularly when low land usage is required. They can combine BOD, solids and ammoniacal nitrogen removal and can be utilised at both secondary and tertiary stages of wastewater treatment. Media selection is critical in the design and operation of BAFs to achieve effluent quality requirements. Two size ranges, 1.5-3.5 and 2.5 4.5 mm, of a foamed clay called StarLight C were used in pilot-scale reactors. Both performed well as BAF media, with reactor loads up to 12 kg COD m(-3) d and 4 kg suspended solids m(-3) d (based on working volumes). The most consistent effluent was obtained using the smaller medium since, at flow rates above 0.41 min(-1), the BAF using the larger medium produced an effluent containing more than 20 mg l(-1) of suspended solids for over 30 min after backwashing. Up to 70% longer run times, as determined by reaching a set head loss, were recorded for the BAF containing the larger rather than the smaller medium. Additionally, the development of pressure above the smaller medium filter bed tended to be logarithmic rather than linear. Reactor profiles indicated that suspended solids removal did not occur over the full 2.3 m depth of the columns. The BAF containing the smaller medium utilised a mean depth of 1.7 +/- 0.3 m, whereas a mean depth of 2.1 +/- 0.3 m was used by the larger medium BAF. Both the head loss development data and the suspended solids removal profiles indicated that the smaller medium BAF was underperforming as a filter. PMID- 11394788 TI - The fate of xenobiotic organic compounds in wastewater treatment plants. AB - The effective operation of wastewater treatment plants plays an important role in minimising the release of xenobiotic compounds into the aquatic environment. Considerable effort has been expended in developing models to quantify the overall removal and fate of these compounds in biological treatment plants. A synthesis and modification of these approaches has been made and a generalised fate model for organic compounds in an activated sludge plant is presented. The influence of the different removal mechanisms, such as sorption, volatilisation and advection for chemicals with different physico-chemical properties is investigated and the important role of biotransformation is discussed. The effect of some operating parameters has been found to have an important influence upon the concentration of xenobiotic released in the sludges and final effluent. This may have significance for a wide range of ecotoxic compounds and in particular the class of compounds increasingly recognised as having the potential to disrupt endocrine activity in some aquatic vertebrates. PMID- 11394789 TI - Dynamics of diffuse pollution from US southern watersheds. AB - To understand the effects of diffuse pollution information on the source of pollutants, quantities in transport, mode of transport, transient nature of the pollution event, and most importantly, a consideration of remediation efforts need to be known. For example, water quality research in the Yazoo Basin uplands in Mississippi has shown sediment loads from a conventional-till upland soybean watershed to be about 19,000kg/ha/yr, and responsible for 77-96% of P and N in transport. In contrast, sediment loads from a comparable no-till soybean watershed were only 500 kg/ha/yr. transporting about 31% of P and N in transport. Sediment loads from a nearby forested area were low, about 200 kg/ha/yr, but responsible for about 47-76% of P and N in transport. Transient pollution events are responsible for the transport of large quantities of sediment, nutrients, and pesticides; in some storm events nearly the annual load. Best management practices (BMPs) must be designed to remediate diffuse pollution and the transient nature of pollution events which can have a profound effect on the ecological health of steams and reservoirs. PMID- 11394790 TI - A simple system to rapidly monitor activated sludge health and performance. AB - A set of four assays designed to rapidly measure the health and biodegradative performance of pulp and paper mill activated sludges was developed. Three of the assays are specific oxygen uptake rates (SOURs) that measure the normal "working" aeration tank BOD (biochemical oxygen demand) removal rate (SOURAT), a near maximum BOD removal rate (SOURNMAX), and a rate (SOURTOX) used in combination with the SOURNMAX to indicate the presence of toxic or inhibitory substances. The fourth assay is the specific adenosine triphosphate (SATP) content of the sludge, used as a measure of its viable cell content. Fresh biomass (sludge) samples from one laboratory reactor and four mill biotreatment systems were fed raw mill effluents and used to evaluate the four-assay set. The SOURAT values of all systems were 10-40% of their SOURNMAX values: thus the SOURAT:SOURNMAX ratios indicate that each system's free biodegradative capacity was far greater than its operating rate. It was demonstrated using phenol that the SOURNMAX:SOURTOX ratio can indicate the presence of substances toxic or inhibitory to the biomass. The results also indicated that the SOURNMAX is a much better indicator of improving or worsening sludge performance and capacity than the SOURAT. SATP was shown to be a useful monitor of the proportion of viable cells in an activated sludge and a toxicity indicator complementary to the SOURNMAX:SOURTOX ratio and similar in principle to the commercial Microtox toxicity test. This four-assay set was also applied to three practical situations: (a) at-mill monitoring of a biotreatment system; (b) effects of cold storage on biomass; and (c) effects of decreased BOD loading on biomass. PMID- 11394791 TI - Removal of easily biodegradable organic compounds by drinking water biofilms: analysis of kinetics and mass transfer. AB - This paper evaluates the rate of utilization of easily biodegradable organic compounds by drinking water biofilms. Tap water, which had been filtered through biologically active granular activated carbon, was used as an innoculum for biofilm growth in annular reactors (ARs). Synthetic cocktails of easily biodegradable material in the concentration range of 50-2,000 mgC/m3 were used as substrate for biofilm growth. Influent and effluent aggregate concentrations of biodegradable organic matter (BOM) were calculated by adding the measurable BOM components on a mass carbon basis. The aggregate BOM values were used for calculating the observed Damkohler number and Theile modulus (based on a reaction rate per unit surface area), which were used to determine whether external or internal mass transfer limited BOM removal. For all of the experimental trials, it was shown that neither external nor internal mass transfer limited BOM removal. Because the biofilms in this research are thin and the fact that mass transfer is not limiting, it was assumed that the bulk BOM concentration was approximately equal to the average BOM concentration in the biofilm. A linear model was obtained for the aggregate BOM flux and the product of the effluent BOM concentration and the biofilm density. The slope or the areal biodegradation rate (ka) for the aggregate BOM was 0.033 m/h, as determined through a linear regression. PMID- 11394792 TI - Dissolved organic carbon losses from grazed grasslands under different management regimes. AB - Dissolved organic matter (DOM) is fundamental to many biogeochemical processes in soils and natural waters. Despite the large number of studies reporting on DOM losses from forest soils and in surface waters there is little published data on exports from managed grasslands. The objective of our study was to determine the extent of short-term exports of dissolved organic carbon (DOC) from managed grazed grasslands and to evaluate the influence of fertilizer management and drainage regime. DOC discharged from grazed grassland plots, with a range of management strategies. was determined over 2 months. Total export varied from 42 to 118 kgCha(-1), and was greater from some plots than literature estimates for annual losses from all catchment types. There was a significant (P = 0.048) positive correlation between DOC export and rates of nitrogen application for treatments with no artificial drainage. Increased dry matter production arising from increased fertilizer-N inputs is suggested as an important factor in this relationship. DOC export was significantly (P = 0.032) reduced by artificial drainage and adsorption of DOC to soil surfaces and the restriction of decomposition due to waterlogging are suggested as two possible explanations. PMID- 11394793 TI - Technical note the pattern of ClO2 stabilized by Na2CO3/H2O2. AB - In this paper, the existing patterns of carbonate and ClO2 in the so-called "stabilized chlorine dioxide" solution which is stabilized by stabilizers Na2CO3 and H2O2 are analyzed. Meanwhile, thermostability, UV absorption spectrum, specificity of the paper chromatogram, microstructure and ionic chromatogram of this solution were studied, and contrasted with pure NaClO2 and ClO2. The results show that ClO2 in "stabilized chlorine dioxide" solution exists in the form of chlorite ClO2-, carbonate exists in the form of bicarbonate. Therefore, it is considered that "stabilized chlorine dioxide" solution is mixed solution of ClO2- and HCO3-, its pH being 8.5. PMID- 11394794 TI - Independent review group calls for greater transparency in the dispensing of medicines. PMID- 11394795 TI - FMD results in 70 per cent fall in submissions to VLA regional laboratories. PMID- 11394796 TI - Jugular venous emboli of brain tissue induced in sheep by the use of captive bolt guns. AB - Emboli of central nervous tissue were detected in the jugular venous blood of two of 15 sheep stunned with a conventional cartridge-operated captive bolt gun and in two of 15 sheep stunned with a pneumatically activated gun. No emboli were detected in arterial blood from these sheep or in venous blood from sheep stunned electrically. Emboli from an animal with BSE could transmit the disease to people. PMID- 11394798 TI - Postmortem findings in captive cane rats (Thryonomys swinderianus) in Gabon. AB - The causes of morbidity and mortality in a population of cane rats (Thryonomys swinderianus) on an experimental farm in Gabon were monitored for 21 months; 94 of 546 animals (17.2 per cent) died and were examined postmortem, and complementary laboratory examinations were carried out on 23 of the cases together with samples from 13 other animals kept elsewhere in similar conditions. Twenty-six (28 per cent) of the deaths occurred in preweaned kits, 40 (42 per cent) in subadults and 28 (30 per cent) in adults. The average monthly mortality was 2.5 per cent. Trauma was responsible for 29 of the deaths, 12 were due to septicaemia, 10 to primary respiratory lesions, five to digestive disorders, four to urinary lesions, three to reproductive problems and three to other causes; no diagnosis could be reached in 28 cases. PMID- 11394797 TI - Value of the pinnal-pedal reflex in the diagnosis of canine scabies. AB - The potential value of the pinnal-pedal scratch reflex as an aid to diagnosing canine scabies was assessed in 588 dogs with skin disease. The reflex was assessed by vigorously rubbing the tip of one earflap on to the base of the ear for five seconds, and it was considered positive if the ipsilateral hind leg made a scratching movement. A diagnosis of scabies was based on the dog's history, a physical examination and either positive skin scrapings or the complete resolution of pruritus and dermatitis after treatment with ivermectin or milbemycin, with no relapse for at least 12 months. Scabies was diagnosed in 55 of the dogs, allergic skin disease in 463, and 70 had other miscellaneous skin diseases. There was a positive pinnal-pedal scratch reflex in 45 (82 per cent) of the 55 dogs with scabies. Forty (73 per cent) of the dogs with scabies had pinnal dermatitis, and 36 (90 per cent) of these had a positive pinnal-pedal scratch reflex. There was a positive pinnal-pedal scratch reflex in 33 (6.2 per cent) of the other 533 dogs. On the basis of these results, the specificity of testing for scabies by the pinnal-pedal scratch reflex was 93.8 per cent, and the sensitivity was 81.8 per cent The test's positive predictive value was 0.57 and its negative predictive value was 0.98. PMID- 11394800 TI - Experimental infection of six-month-old sheep with Escherichia coli O157:H7. PMID- 11394799 TI - Plasma atrial natriuretic peptide concentrations in hypocalcaemic cows. PMID- 11394801 TI - Use of acupuncture for the treatment of idiopathic facial nerve paralysis in a dog. PMID- 11394802 TI - Occurrence of Escherichia coli O157 in reindeer (Rangifer tarandus). PMID- 11394803 TI - 50,XY gonadal dysgenesis (Swyer's syndrome) in a female river buffalo (Bubalus bubalis). PMID- 11394804 TI - Isolation of Candida krusei from a case of bovine bronchopneumonia in a one-year old heifer. PMID- 11394805 TI - FMD control strategies. PMID- 11394806 TI - FMD control strategies. PMID- 11394807 TI - FMD control strategies. PMID- 11394808 TI - FMD diagnosis. PMID- 11394809 TI - Working together to fight emerging diseases. PMID- 11394810 TI - Executive summary: AVMA comments on the AAVSB draft model state veterinary practice act. American Association of Veterinary State Boards. PMID- 11394811 TI - Updated information for treatment of fungal infections in cats and dogs. PMID- 11394812 TI - Believes decreased dietary intake would reduce flatulence in dogs. PMID- 11394813 TI - Thinks grading of mast cell tumors is inconsistent. PMID- 11394814 TI - Another view on educational models. PMID- 11394815 TI - Another reader opposing thoracic compression for avian euthanasia. PMID- 11394817 TI - The presence of mind: on reunifying the animal mind and body. PMID- 11394816 TI - Questions whether clients were properly informed about treatment protocol. PMID- 11394818 TI - An insight into the AVMA Guidelines for Complementary and Alternative Veterinary Medicine. PMID- 11394819 TI - AVMA Guidelines for Complementary and Alternative Veterinary Medicine. PMID- 11394820 TI - A community approach to dog bite prevention. PMID- 11394821 TI - What is your diagnosis? Severe soft tissue swelling over the brachium and antebrachium and an avulsion-type fracture of the olecranon. PMID- 11394822 TI - Generation and gender--a critical mix. PMID- 11394823 TI - Evaluation of nutraceuticals, dietary supplements, and functional food ingredients for companion animals. PMID- 11394824 TI - Savvy buying and selling of veterinary practices. PMID- 11394825 TI - Noncompetition agreements--some recent wrinkles. PMID- 11394826 TI - Practitioner report involving intravenous use of vitamin K1 prompts label review and revision. PMID- 11394827 TI - Development of a complete electronic medical record in an academic institution. PMID- 11394828 TI - The paperless practice. PMID- 11394829 TI - Serologic evidence of Rickettsia akari infection among dogs in a metropolitan city. AB - OBJECTIVE: To determine whether dogs in New York, NY are naturally infected with Rickettsia akari, the causative agent of rickettsialpox in humans. DESIGN: Serologic survey. ANIMALS: 311 dogs. PROCEDURE: Serum samples were obtained from dogs as a part of a study on Rocky Mountain spotted fever and borreliosis or when dogs were examined at area veterinary clinics for routine care. Dog owners were asked to complete a questionnaire inquiring about possible risk factors at the time serum samples were obtained. Samples were tested for reactivity to spotted fever group rickettsiae by use of an enzyme immunoassay (EIA). Twenty-two samples for which results were positive were tested by use of an indirect immunofluorescence antibody (IFA) assay followed by confirmatory cross-absorption testing. RESULTS: Results of the EIA were positive for 24 (7.7%) dogs. A history of tick infestation and increasing age were significantly associated with whether dogs were seropositive. Distribution of seropositive dogs was focal. Seventeen of the 22 samples submitted for IFA testing had titers to R rickettsii and R akari; for 11 of these, titers to R akari were higher than titers to R rickettsii. Cross absorption testing indicated that in 6 of 7 samples, infection was caused by R akari. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that dogs can be naturally infected with R akari. Further studies are needed to determine the incidence of R akari infection in dogs, whether infection is associated with clinical illness, and whether dogs can serve as sentinels for human disease. PMID- 11394831 TI - Prevalence of behavioral changes associated with age-related cognitive impairment in dogs. AB - OBJECTIVE: To determine the prevalence of age-related behavioral changes, namely impairment, in a randomly chosen population of dogs. DESIGN: Age-stratified cohort study. ANIMALS: 97 spayed female and 83 castrated male dogs that were 11 to 16 years old. PROCEDURE: Data on possible impairment in 4 behavioral categories (ie, orientation in the home and yard, social interaction, house training, and sleep-wake cycle) linked to cognitive dysfunction were obtained from dog owners, using a structured telephone interview. Hospital records of dogs had been screened to exclude dogs with dysfunction in organ systems that may cause behavioral changes. Dogs with behavioral impairment were those with > or = 2 signs of dysfunction within a category. Dogs with impairment in 1 category were considered mildly impaired and those with impairment in > or =2 categories were considered severely impaired. RESULTS: Age by sex interactions for dogs with impairment in any category were not significant, and, therefore, data on castrated males and spayed females were pooled for analyses across ages. The prevalence of age-related progressive impairment was significant in all categories. The percentage of 11- to 12-year-old dogs with impairment in > or = 1 category was 28% (22/80), of which 10% (8/80) had impairment in > or = 2 behavioral categories. Of 15- to 16-year-old dogs, 68% (23/34) had impairment in > or =1 category, of which 35% (12/34) had impairments in > or = 2 categories. There were no significant effects of body weight on the prevalence of signs of dysfunction in the behavioral categories. CONCLUSIONS AND CLINICAL RELEVANCE: Data collected provide estimates of the prevalence of various degrees of age related behavioral changes associated with cognitive dysfunction in dogs. Age related behavioral changes may be useful indicators for medical intervention for dogs with signs of cognitive impairment. PMID- 11394830 TI - Evaluation of piroxicam for the treatment of oral squamous cell carcinoma in dogs. AB - OBJECTIVE: To evaluate the use of piroxicam for the treatment of oral squamous cell carcinoma in dogs. DESIGN: Prospective case series. ANIMALS: 17 dogs with measurable oral squamous cell carcinoma. PROCEDURE: Dogs were treated with piroxicam at a dosage of 0.3 mg/kg (0.14 mg/lb) of body weight, PO, every 24 hours until progressive disease or unacceptable signs of toxicosis developed or the dog died. RESULTS: One dog had a complete remission (maxillary tumor), and 2 dogs had partial remissions (lingual tumor and tonsillar tumor). An additional 5 dogs had stable disease, including 1 with a maxillary tumor, 2 with mandibular tumors, and 2 with tonsillar tumors. Variables associated with tumor response were not identified. Median and mean times to failure for the 3 dogs that had a remission were 180 and 223 days, respectively. Median and mean times to failure for the 5 dogs with stable disease were 102 and 223 days, respectively. Time to failure was positively associated with tumor response and negatively associated with tumor size. One dog had mild adverse gastrointestinal tract effects that resolved with the addition of misoprostol to the treatment regimen. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that piroxicam may be useful in the treatment of dogs with oral squamous cell carcinoma; response rate was similar to that reported for other cytotoxic treatments. Larger-scale studies are warranted to determine what role piroxicam may have, alone or in combination with other treatments, for the treatment of dogs with oral squamous cell carcinoma. PMID- 11394832 TI - Predicting behavioral changes associated with age-related cognitive impairment in dogs. AB - OBJECTIVE: To monitor the progression of age-related behavioral changes in dogs during a period of 6 to 18 months and to determine whether signs of dysfunction in any of 4 behavioral categories can be used to predict further impairment. DESIGN: Age-stratified cohort study. ANIMALS: 63 spayed female and 47 castrated male dogs 11 to 14 years of age. PROCEDURE: Data were collected from randomly selected dog owners who were interviewed by telephone twice at a 12- to 18-month interval; data were included if the dog had lived > or = 6 months between interviews. The interview focused on signs of impairment in the following behavioral categories: orientation in the home and yard, social interactions with human family members, house training, and the sleep-wake cycle. Dogs were determined to have impairment in 0 behavioral categories (on the basis of < or = 1 sign for each category), impairment in 1 category (> or = 2 signs of dysfunction in that category), or impairment in > or = 2 categories. RESULTS: Between interviews, 22% (16/73) of dogs that did not have impairment in a category at the time of the first interview developed impairment in that category by the time of the second interview. Forty-eight percent (13/27) of dogs that had impairment in 1 category at the time of the first interview developed impairment in > or = 2 categories by the time of the second interview and were significantly more likely to develop impairment in > or = 2 categories, compared with dogs that initially had impairment in 0 categories. Dogs with 1 sign of dysfunction in orientation were significantly more likely to develop impairment in that category, compared with dogs that had 0 signs of dysfunction in orientation. CONCLUSIONS AND CLINICAL RELEVANCE: Age-related behavioral changes in dogs are progressive. Clinicians should consider trying to predict which dogs are most likely to become progressively impaired during the subsequent 6 to 18 months. PMID- 11394833 TI - Ultrasound-guided percutaneous drainage for treatment of pyonephrosis in two dogs. AB - Pyonephrosis refers to suppurative destruction of the parenchyma of the kidney with complete or nearly complete loss of renal function. In dogs, nephrectomy is still the most common treatment for pyonephrosis; however, in the present report, a method for percutaneous ultrasound-guided drainage of the renal pelvis in dogs with pyonephrosis that does not require local or general anesthesia was described, and results of the procedure in 2 dogs were reported. Briefly, dogs were positioned in lateral recumbency with the affected side up, and skin overlying the affected kidney was aseptically prepared. The dilated renal pelvis was punctured percutaneously, under ultrasound guidance, with a 22-gauge needle, and a sample of material was obtained for analysis. The needle was then replaced with an IV catheter, and as much pus as possible was removed from the renal collecting system. A povidone iodine solution was then used to lavage the renal pelvis. Ultrasound-guided drainage and lavage of the renal pelvis was repeated daily until the renal pelvis was so small that it could no longer be punctured. Both dogs recovered and were reported by the owners to be healthy after the procedure. PMID- 11394834 TI - Scedosporium prolificans infection associated with arthritis and osteomyelitis in a horse. AB - Scedosporium prolificans was associated with arthritis and degenerative osteomyelitis in a 6-year-old Thoroughbred racehorse. The horse was suspected to have an inflammatory lesion of the interosseous tendon, but treatment had resulted in only a minimal response. Shortly after diagnostic arthrocentesis of the left metacarpophalangeal joint was performed, the joint became severely swollen, and radiography of the area revealed lysis of the distal end of the third metacarpal bone, the proximal sesamoid bones, and the proximal end of the proximal phalanx. The horse did not respond to treatment and was euthanatized. At necropsy, severe erosive arthritis and degenerative osteomyelitis of the left metacarpophalangeal joint were seen. Swab specimens of the ulcerated lesions of the articular cartilage were submitted for microbial culture, and Scedosporium prolificans was isolated. Scedosporium prolificans is a newly recognized opportunistic fungal pathogen of humans and animals. In humans, S prolificans typically causes focal locally invasive infections that primarily involve musculoskeletal tissues; most often, infection is a result of penetrating trauma or surgical incision. In immunocompromised patients, fatal disseminated infection can occur. The fungus is resistant to almost all currently available antimycotic agents. PMID- 11394836 TI - "Be prepared". PMID- 11394835 TI - Evaluation of the embryo transfer procedure proposed by the International Embryo Transfer Society as a method of controlling vertical transmission of Neospora caninum in cattle. AB - OBJECTIVE: To evaluate efficacy of embryo transfer into seronegative recipients, using the procedure proposed by the International Embryo Transfer Society (IETS), for preventing vertical transmission of Neospora caninum in cattle. DESIGN: Prospective clinical trial. ANIMALS: 87 recipient cows and heifers and their embryo transfer calves from 22 donors originating from 9 dairy herds. PROCEDURE: Neospora caninum serologic status of donors and recipients was determined before collection and transfer of embryos. Viable embryos were washed and treated with trypsin. Recipients in experimental groups A (n = 50) and B (29) were seronegative and received embryos from seropositive and seronegative donors, respectively. Recipients in group C (n = 8) were seropositive and received embryos from seronegative or seropositive donors. Antibody titers against N caninum were determined monthly during pregnancy in recipients and in calf blood samples collected at birth. Tissues collected from stillborn calves and aborted fetuses were analyzed histologically and by immunohistochemical (IHC) methods. RESULTS: 76 calves and 11 fetuses and stillborn calves were examined. All calves from groups A and B were seronegative (n = 70) or lacked evidence of infection by use of tissue analysis (9). In group C, 5 of 6 calves were seropositive at birth, and IHC results were positive for 1 of 2 calves. Vertical transmission rate was significantly lower in groups A and B (0%) than in group C (75%). CONCLUSION AND CLINICAL RELEVANCE: Embryo transfer into seronegative recipients, using the procedure proposed by IETS, is an effective way to prevent vertical transmission of N caninum. Results provide support for pretransfer testing of all embryo transfer recipients. PMID- 11394837 TI - Patient communication--the hidden procedure in the management of glaucoma. PMID- 11394838 TI - Myopia in Singapore kindergarten children. AB - PURPOSE: The purpose of this study was to examine whether close-up work was related to myopia in Singapore kindergarten children. METHODS: One hundred twenty eight children, ages 3 to 7 years, from a kindergarten in Singapore were examined by cycloplegic autorefraction. The parents also completed a questionnaire on the different types of close-up work activities each child was engaged in, socioeconomic status, and parental history of myopia. RESULTS: Myopic children spent 3.0 hours per day (median) on close-up work activity, while nonmyopic children spent 2.0 hours per day (median) on close-up work activity. The prevalence of myopia in the sample was 8.6%. CONCLUSION: Close-up work activity was not related to myopia in pre-school children. PMID- 11394839 TI - Prescribing patterns of optometrists newly licensed in therapeutic drug use in New York State. AB - PURPOSE: As part of the New York State law authorizing optometrists to use Therapeutic Pharmaceutical Agents (TPAs), the New York State Education Department commissioned a study to monitor and assess the prescribing patterns and competence of optometrists newly licensed to use TPAs. This report summarizes the findings of that study. METHODS: The database was obtained from more than 11,000 Patient Data Reporting Forms (PDRFs) returned by 535 optometrists, listing diagnosis, type(s) of medication(s) prescribed, treatment outcome, and possible adverse effects. RESULTS: Of the more than 11,000 forms returned by 535 optometrists, 8,936 were first visits, with 5,828 follow-up visits scheduled. For all reported initial visits, only 89 patients required ophthalmological referral for secondary or tertiary care. No adverse drug reactions requiring intervention were reported. The most-frequent diagnoses treated by optometrists were related to allergies, superficial trauma, conjunctivitis (unspecified), and then blepharitis. The most commonly prescribed ophthalmic preparations as actually reported on the PDRFs were Polytrim (trimethoprim/ polymixin B), tobramycin (brand name and generic), TobraDex (tobramycin/dexamethasone), and Livostin (levocabastine). CONCLUSIONS: When analyzed according to diagnosis, the data indicate that the prescribing patterns of the prescribing optometrists adhered to currently accepted clinical guidelines. Based on the available data and the lack of observed side effects, the use of topical medications to treat anterior segment disease by optometrists in New York State appears to be safe, and should be of benefit to patients who seek care from optometrists. PMID- 11394840 TI - Suprasellar squamous papillary craniopharyngioma: a case report. AB - BACKGROUND: Craniopharyngiomas are benign neoplasms that are usually suprasellar in origin. They are considered a type of pituitary tumor that can cause hyposecretion of anterior pituitary hormones or hypopituitarism. Clinically, they may cause visual and endocrine deficiencies similar to pituitary adenoma. Histopathologically, they are distinct from pituitary adenoma and are generally classified as either adamantinomatous or squamous papillary. Adamantinomatous tumors are more common in children, whereas the squamous papillary type is found almost exclusively in adults. CASE REPORT: A 45-year-old white man came to our clinic with a history of decreased vision in his left eye for the previous five months that fluctuated. Magnetic resonance imaging of the brain revealed a suprasellar multilobular mass, which was distinguishable from the pituitary gland. The patient underwent a right frontotemporal craniotomy for resection of the tumor. Histopathological analysis showed a benign squamous epithelial-lined mass consistent with squamous papillary craniopharyngioma. CONCLUSION: Craniopharyngiomas are generally considered to be adamantinomatous or squamous papillary in origin. Recently, magnetic resonance imaging has been used to distinguish between these two types of tumors, although clinically no significant difference has been found with respect to resectability. Clinical presentation, treatment options, recurrence, and differential diagnosis of craniopharyngiomas are discussed. PMID- 11394841 TI - Retinal arteriovenous communication. AB - BACKGROUND: Disturbances in development of the embryonic vascular system, though uncommon, may cause formation of retinal arteriovenous communications (RAC). Such anomalies may also involve the intracranial, orbital, and maxillofacial blood vessels. Field loss, neovascular glaucoma, optic atrophy, Parinaud syndrome, hemiparesis, hemiplegia, and proptosis may be associated with RAC. CASE REPORT: A 27-year-old woman was referred for photography of a vascular anomaly in her left eye. I noted a large arteriovenous malformation joining major temporal branches of the left central retinal artery and vein. External and internal examination of both eyes was otherwise unremarkable. Visual-field testing revealed nasal field disturbances for the left eye, but none for the right eye. CT scan showed no obvious intracranial abnormalities. CONCLUSIONS: Visual-field testing may show scotomas associated with retinal arteriovenous communications. It is possible for patients with RAC to have no associated vascular abnormalities elsewhere in the circulation of the head. PMID- 11394842 TI - Documented acquired asteroid hyalosis in a case of early diagnosed diabetes mellitus. AB - BACKGROUND: There have been many reports in the literature of the possible linkage of asteroid hyalosis (AH) to diabetes mellitus (DM). The controversy regarding an association between AH and DM has been one of the longest disputes in the ophthalmic literature. Here we present a case in which AH developed in a patient followed for 9 years after being diagnosed with DM. CASE REPORT: The patient had been examined on nine occasions (since his initial visit in June 1989) and asteroid hyalosis was not discovered until July 1996, when he came in with newly diagnosed diabetes mellitus. This suggests there may well be an association of asteroid hyalosis and diabetes mellitus. DISCUSSION: The patient in this case had a number of risk factors for the development of this ocular condition. He had a long history of systemic arterial hypertension, which has been reported to be linked to the formation of AH. He also had a chronic case of cystoid macular edema, which indicated a vascular compromise to the retinal vessels in the posterior pole, and this leakage may be responsible for serous constituents leaking into the vitreous, which may have caused AH. CONCLUSIONS: This may be the first time in the reported literature that AH was found to occur in a previously normal-appearing vitreous, which was documented over a 9-year period. We would suggest that asteroid hyalosis may be secondary to some form of vasculopathy in many incidences and that diabetes mellitus is one of the conditions that may be associated with the formation of AH. PMID- 11394843 TI - An atypical case of choroidal osteomas. AB - BACKGROUND: A choroidal osteoma is a benign choroidal tumor It usually occurs in young, healthy women and is typically unilateral and located in the juxtapapillary region. CASE REPORT: A case of bilateral choroidal osteomas occurring in a 32-year-old woman is reported. The visual symptoms coincided with her third pregnancy. The choroidal osteoma in her right eye was elevated with blurred margins and had signs of subretinal fluid. The lesion in her left eye was flat and dry. Both were located superior temporal to the macula. Fluorescein angiography and indocyanine green angiography showed evidence of subretinal fluid in her right eye. CONCLUSIONS: The atypical features of the osteoma in her right eye include its height, evidence of diffuse leakage of subretinal fluid with no definable neovascular membrane, and its possible growth associated with pregnancy. In addition, the location, indistinct borders, and bilateral presentation are not characteristic clinical features of choroidal osteomas. PMID- 11394844 TI - An alternative source of referrals. PMID- 11394845 TI - Optometry on the road. PMID- 11394846 TI - Target your market. PMID- 11394847 TI - Sesquiterpene hydrocarbons with trifarane backbone in the liverwort Trocholejeunea sandvicensis. AB - The hydrocarbon fraction of the liverwort Trocholejeunea sandvicensis was investigated. Four new sesquiterpene hydrocarbons including the trifaranes (-) trifara-9,14-diene (6) and (-)-3,7-di-epi-3,7-trifara-9,14-diene (7), as well as (+)-sandvicene (8), a sesquiterpene with uncommon skeleton, and the rearranged trifarane sesquiterpene (+)-neotrifaradiene (9) were isolated and their structure elucidated. A biogenetic pathway for the four compounds is proposed, with nerolidol as their common precursor. PMID- 11394848 TI - Pregnene derivatives from Solenostemma argel leaves. AB - Two new pregnene derivatives 14beta-15alpha-dihydroxy-delta4pregnene-3,20 dione and 3beta-14beta,15alpha-16alpha hydroxy-20-oxo-delta5pregnene-tetra-ol, in addition to alpha- and beta-amyrin and beta-sitosterol, were isolated from Solenostemma argel leaves. The structures were established by extensive spectral analysis as well as comparison with reference materials. PMID- 11394850 TI - Furanoditerpenoid lactones from the seeds of Caesalpinia minax Hance. AB - Two furanoditerpenoid lactones, were isolated from the seeds of Caesalpinia minax Hance. Their structures and stereochemistry have been established by spectral data and single crystal X-ray analysis. PMID- 11394849 TI - An eudesmanolide and a carotane from Ferula sinaica. AB - Re-examination of the chemical constituents of the leaves of Ferula sinaica afforded a new eudesmanolide and a new carotane. The structures were elucidated by spectroscopic methods. PMID- 11394851 TI - Guaianolides from Cichorium intybus and structure revision of Cichorium sesquiterpene lactones. AB - The isolation and structure elucidation of a new lactucopicrin derivative from Cichorium intybus is described, together with the revised structures of several sesquiterpene lactones previously isolated from Cichorium species. The known eudesmanolide magnolialide and the known guaianolide ixerisoside D, reported for the first time from the plant material, along with the previously known chicory sesquiterpene lactones have been also isolated and identified. PMID- 11394852 TI - (S)-12-hydroxygeranylgeraniol-derived diterpenes from the brown alga Bifurcaria bifurcata. AB - Four novel diterpenes were isolated from the brown alga Bifurcaria bifurcata collected off the Atlantic coast from Morocco, and their structures established by spectral and chemical methods. These compounds are acyclic diterpenes derived from (S)-12-hydroxygeranylgeraniol. One of them is its dehydration product at C 12, while the others are its oxidation derivatives: the methyl ester of the acid at C-1 and two stereoisomers (Z and E) of the aldehyde at C-1. These results are discussed from a chemotaxonomic point of view. PMID- 11394853 TI - Diterpenes from the bark and seeds of Colophospermum mopane. AB - The CHCl3 extract of the bark and seeds of Colophospermum mopane gave three new diterpenes, dihydrogrindelic acid and dihydrogrindelaldehyde and methyl labd-13E en-15-oate. The structures were assigned on the basis of detailed spectroscopic data analyses. The aldehyde showed significant cytotoxic activity against a human breast cancer cell line. PMID- 11394854 TI - Flavonoids from Aconitum napellus subsp. neomontanum. AB - Three flavonol glycosides quercetin 7-O-(6-trans-caffeoyl)-beta-glucopyranosyl-(1 ->3)-alpha-rhamnopyranoside-3-O-beta-glucopyranoside (1), kaempferol 7-O-(6-trans caffeoyl)-beta-glucopyranosyl-(1-->3)-alpha-rhamnopyranoside-3-O-beta glucopyranoside (2), and kaempferol 7-O-(6-trans-p-coumaroyl)-beta-glucopyranosyl (1-->3)-alpha-rhamnopyranoside-3-O-beta-glucopyranoside (3), together with the known beta-3,4-dihydroxyphenethyl beta-glucopyranoside, were isolated from the flowers of Aconitum napellus subsp. neomontanum. Their structures were elucidated by spectroscopic methods, including 2D NMR spectral techniques. PMID- 11394855 TI - Quercetin 3,3',4'-tri-O-beta-D-glucopyranosides from leaves of Eruca sativa (Mill.). AB - Three new quercetin 3,3',4'-tri-O-beta-D-glucopyranosides isolated from leaves of Eruca sativa (Mill.) were identified as quercetin 3,3',4'-tri-O-beta-D glucopyranoside, quercetin 3'-(6-sinapoyl-O-beta-D-glucopyranosyl)-3,4'-di-O-beta D-glucopyranoside and quercetin 3-(2-sinapoyl-O-beta-D-glucopyranosyl)-3'-(6 sinapoyl-O-beta-D-glucopyranosyl)-4'-O-beta-D-glucopyranoside. The structures were established by one- and two-dimensional 1H and 13C NMR spectra as well as b PMID- 11394856 TI - Natural and synthetic benzophenones: interaction with the cytosolic binding domain of P-glycoprotein. AB - A benzophenone glycoside has been isolated from Davallia solida. Its structure was elucidated by chemical and spectral means as 4-O-beta-D-glucopyranosyl-2,6,4' trihydroxybenzophenone. It bound with moderate affinity to the purified C terminal cytosolic domain of P-glycoprotein, but the binding affinity was 6- to 10-fold increased for its aglycone derivative and other related benzophenones. PMID- 11394857 TI - Two flavonoids and other compounds from the aerial parts of Centaurea bracteata from Italy. AB - The flowering aerial parts of Centaurea bracteata Scop. (Asteraceae) have been studied for the first time. Nineteen compounds were isolated and identified, namely a sterol glucoside, two phenolic acids, three quinic acid derivatives, and 13 flavonoids, two of which, are new natural products. Structural elucidation was performed mainly by mean of FABMS, 1D and 2D NMR spectroscopy. PMID- 11394858 TI - The flavonoids of leek, Allium porrum. AB - A phytochemical investigation of the extracts obtained from bulbs of leek. Allium porrum L. has led to the isolation of five flavonoid glycosides based on the kaempferol aglycone. Two of them are new compounds and have been identified as kaempferol 3-O-[2-O-(trans-3-methoxy-4-hydroxycinnamoyl)-beta-D-galactopyranosyl] (1-->4)-O-beta-D-glucopyranoside, and kaempferol 3-O-[2-O-(trans-3-methoxy-4 hydroxycinnamoyl)-beta-D-glucopyranosyl]-(1-->6)-O-beta-D-glucopyranoside, on the basis of spectroscopic methods, including 2D NMR. The isolated compounds have been evaluated for their human platelet anti-aggregation activity. PMID- 11394859 TI - Flavones and phenylpropenoids in the surface exudate of Psiadia punctulata. AB - Three flavones, 5,7-dihydroxy-2',3',4',5'-tetramethoxyflavone, 5,4'-dihydroxy 7,2',3',5'-tetramethoxyflavone, and 5,7,4'-trihydroxy-2',3',5'-trimethoxyflavone were isolated from the leaf exudate of Psiadia punctulata, together with the previously reported 5-hydroxy-7,2',3',4',5'-pentamethoxyflavone and 5,7,3' trihydroxy-2',4',5'-trimethoxyflavone. The two phenylpropenoids, Z-docosyl-p coumarate and E-docosyl-p-coumarate were also isolated. The structures were determined on the basis of spectroscopic evidence. PMID- 11394860 TI - Kaempferol triosides from Reseda muricata. AB - A flavonoid trioside and its coumaryl ester together with seven known flavonoids and five phenolic acids were isolated from the leaves of Reseda muricata. Their structures were elucidated by spectroscopic methods including UV, FAB MS, 1H, 13C and 2D-NMR, DEPT, HMBC and HMQC experiments. The two compounds were identified as kaempferol 3-O-beta-D-glucopyranosyl-(1''' --> 2'')-O-alpha-L-rhamnopyranoside 7 O-beta-D-glucopyranoside and kaempferol 3-O-beta-D-glucopyranosyl-(1''' --> 2'') O-alpha-L rhamnopyranoside 7-O-beta-D-(6''''-O-E-coumarylglucopyranoside), respectively. PMID- 11394861 TI - Biflavonoids from Ochna afzelii. AB - The stem bark of Ochna afzelii furnished three biflavonoids among which two are new. Their structures were established from spectroscopic and chemical evidences. PMID- 11394862 TI - Oxanthrone esters from the aerial parts of Cassia kleinii. AB - From the aerial parts of Cassia kleinii two new oxanthrone esters, kleinioxanthrone-1 and kleinioxanthrone-2 have been isolated. Their structures were established as 1,8-dihydroxy-3-methyl-6-methoxy-9(10H)-anthracenone-10 oxydecanoate 1 and 1,8-dihydroxy-3-methyl-9(10H)-anthracenone-10 oxytetradecanoate 2 respectively based on degradative and spectroscopic evidence. PMID- 11394863 TI - Ellagic acid rhamnosides from the stem bark of Eucalyptus globulus. AB - Four ellagic acid rhamnosides were isolated from the stem bark of Eucalyptus globulus. Their structures have been established on the basis of the analysis of their 1H NMR, 13C NMR, HMBC, IR and MS spectral data. The HMBC data of these compounds were most useful for their structure determinations, with these bring determined to be 3-O-methylellagic acid 3'-O-alpha-rhamnopyranoside, 3-O methylellagic acid 3'-O-alpha-3''-O-acetylrhamnopyranoside, 3-O-methylellagic acid 3'-O-alpha-2''-O-acetylrhamnopyranoside, 3-O-methylellagic acid 3'-O-alpha 4''-O-acetylrhamnopyranoside, respectively. Their antioxidant activities were evaluated by measuring the inhibition of lipid peroxidation using rat liver microsomes, with IC50 values of 10.0-14.0 microg/ml. PMID- 11394864 TI - Furomegistines I and II, two furanopyridine alkaloids from the bark of Sarcomelicope megistophylla. AB - Two alkaloids, furomegistine I (1) and furomegistine II (2), were isolated from the bark of Sarcomelicope megistophylla. Their structures have been elucidated on the basis of MS and NMR data. Both belong to the category of furanopyridine alkaloids and should be considered as oxidation products of a furo[2,3 b]quinoline precursor. The two alkaloids exhibited moderate cytotoxic activity. PMID- 11394865 TI - Alkaloids from Haplophyllum tuberculatum. AB - Two new alkaloids, haplotubinone (3) and haplotubine (4), were isolated from the aerial parts of Haplophyllum tuberculatum together with the known lignan diphyllin. The structures of the new alkaloids were established by spectroscopic methods in conjunction with X-ray crystallographic analysis of 3. In addition, the amide N-(2-phenylethyl)-benzamide has been identified in this source for the first time. PMID- 11394866 TI - Methyl chanofruticosinates from leaves of Kopsia flavida Blume. AB - Three new indole alkaloids with methyl chanofruticosinates skeletal system, viz., methyl 12-methoxy-N1-decarbomethoxychanofruticosinate, methyl 12 methoxychanofruticosinate and methyl 11,12-dimethoxychanofruticosinate, in addition to methyl 11,12-methylenedioxy-N1-decarbomethoxychanofruticosinate, have been isolated from the leaves of Kopsia flavida Blume. The structures of these three new indole alkaloids were assigned by NMR spectral data using various 2D techniques. PMID- 11394867 TI - Polyhalogenated homosesquiterpenic fatty acids from Plocamium cartilagineum. AB - We describe the composition of novel polyhalogenated homosesquiterpenic acids from the red alga Plocamium collected during summer on Maltese Islands and Corsica. The compounds, predominantly derivatives with unique groups (bromine, chlorine, and diene), were identified by means of 1H and 13C NMR, MS, IR and UV spectra. PMID- 11394868 TI - Nostocyclamide M: a cyanobacterial cyclic peptide with allelopathic activity from Nostoc 31. AB - A cyclic peptide containing thiazole and oxazole moieties was isolated from Nostoc 31 and its structure determined by chemical degradation detailed NMR and mass spectroscopic analyses. The compound is stereochemically pure and closely related to nostocyclamide in which D-valine is replaced by D-methionine. Therefore, it differs from tenuecyclamide C reported to contain L-methionine. It shows allelopathic activity against Anabaena 7120, but is inactive against grazers. PMID- 11394869 TI - Biochemical analysis of yeast G(alpha) mutants that enhance adaptation to pheromone. AB - The mating-specific heterotrimeric G(alpha) protein of Saccharomyces cerevisiae, Gpa1, negatively regulates activation of the pheromone response pathway both by sequestering G(beta)gamma and by triggering an adaptive response through an as yet unknown mechanism. Previous genetic studies identified mutant alleles of GPA1 that downregulate the pheromone response independently of the pheromone receptor (GPA1E364K), or through a receptor-dependent mechanism (GPA1N388D). To further our understanding of the mechanism of action of these mutant alleles, their corresponding proteins were purified and subjected to biochemical analysis. The receptor-dependent activity of Gpa1N388D was further analyzed using yeast strains expressing constitutively active receptor (Ste2) mutants, and C-terminal truncation mutant forms of Gpa1. A combination of G(alpha) affinity chromatography, GTP binding/hydrolysis studies, and genetic analysis allowed us to assign a distinct mechanism of action to each of these mutant proteins. PMID- 11394870 TI - Homozygosity mapping to chromosome 5p15 of a gene responsible for Hartnup disorder. AB - Hartnup disorder is an autosomal recessive phenotype involving a transporter for monoamino-monocarboxylic acids. Genetic analysis of the mouse model mapped its locus to human chromosome 11q13 (8). We report here the results of linkage analysis in two Japanese first cousin-marriage families. In the first family, the proband had Hartnup disorder and his deceased older brother was reported to have had typical Hartnup symptoms. The younger brother of the proband was shown to have decreased tryptophan absorption by oral loading test. In the second family, a 6-year-old girl, the proband, had specific hyperaminoaciduria. DNA was isolated from either blood samples or umbilical cord stumps. Genome-wide screening by homozygosity mapping was conducted. Taking into account that the older brother was affected and the younger brother was a carrier in the first family, homozygosity mapping (LOD score = 3.55) and GENEHUNTER (LOD score = 3.28) locates the locus of the Hartnup disorder on 5p15. PMID- 11394871 TI - The protective role of HSP90 against 3-hydroxykynurenine-induced neuronal apoptosis. AB - 3-hydroxykynurenine (3HK), an endogenous metabolite of tryptophan in the kynurenine pathway, is a potential neurotoxin in several neurodegenerative disorders. Stabilizing protein structure, heat shock proteins (HSPs) have diverse roles as molecular chaperones to mediate stress tolerance. In the present study, we investigated the possible protective role of HSPs against 3HK induced neuronal cell death. Here we report that 3HK induced in a dose- and time-dependent manner neuronal cell death in neuroblastoma SK-N-SH cells. The cell death showed characteristic apoptotic features such as cell shrinkage, plasma membrane blebbing, chromatin condensation, and nuclear condensation and fragmentation. Furthermore, SK-N-SN cells were protected from 3HK induced cytotoxicity by prior elevation of HSPs expression. Our results show that the protective effect was abolished by HSP90 anti-sense oligonucleotides while not by HSP27 and HSP70 anti sense oligonucleotides. Also, our result shows that HSP90 effectively inhibits caspases activities leading to the apoptosis. These results suggest that 3HK induces apoptosis in neuroblastoma SK-N-SN cells and that HSP90 is major contributing protein component of protection against 3HK induced apoptosis. PMID- 11394872 TI - Photoaffinity labeling of human IRBP with all-trans-retinoic acid. AB - Interphotoreceptor retinoid-binding protein (IRBP), found only in photosensitive tissues, is a large approximately 135-kDa glycoprotein that contains a fourfold repeat structure. IRBP may function as a buffer and prevent retinoid toxicity and retinoid degeneration. Here we asked (i) whether each repeat of IRBP possesses the capability of photo-crosslinking all-trans-retinoic acid (RA), (ii) within Repeat 1 whether a single retinoic acid-binding domain exists, and (iii) whether protease and CNBr digestion of Repeat 1 bound RA indicate the exact location of the binding site. 3H-RA cross-linked to all four repeats, consistent with the current model of multiple binding sites in IRBP. Acetone precipitation was effective in removing unbound 3H-RA. LysC and tryptic digestion of the RA-Repeat 1 detected 18- and 5-kDa bands, respectively. CNBr digestion showed two bands about 9 and 11 kDa in size. Our data suggests a single binding site near positions 151-160 in the center of Repeat 1. PMID- 11394874 TI - Alterations of MAPK activities associated with intestinal cell differentiation. AB - Three distinct groups of mitogen-activated protein kinases (MAPKs) have been identified in mammalian cells (i.e., ERK, JNK, and p38) which play an important role in the differentiation and apoptosis of various cells. The purpose of our present study was to determine MAPK activity and levels associated with sodium butyrate (NaBT)-mediated differentiation and apoptosis in the human colon cancer cell lines Caco-2 and HT29. Intestinal alkaline phosphatase (IAP) activity, a marker of intestinal differentiation, was increased at 48 h after NaBT treatment followed by cell death at 72 h. ERK activity was decreased in differentiated Caco 2 cells either induced with NaBT or allowed to differentiate spontaneously and in HT29 cells treated with NaBT. The combination of the MEK inhibitor, PD98059, with NaBT further increased IAP activity and cell death compared with NaBT alone. In contrast to ERK, JNK1 activity and c-Jun phosphorylation was increased 8 h after NaBT treatment suggesting a role for the JNK pathway in intestinal cell differentiation and apoptosis. p38 activity was increased at 24 and 48 h after NaBT treatment. Taken together, our results suggest that alterations in MAPKs (i.e., ERK inhibition and JNK induction) contribute to the differentiation and apoptotic pathways in intestinal cells. PMID- 11394873 TI - Adaptor protein Lad relays PDGF signal to Grb2 in lung cells: a tissue-specific PDGF signal transduction. AB - Lad was previously identified as an adaptor protein binding to the SH2 domain of Lck (1). Specific detection of Lad mRNA in lung cells, as well as, in T cells led us to investigate the signaling pathways regulating Lad in lung cells. We found that (i) upon PDGF stimulation, Lad expression is induced in lung cells, especially in the bronchial epithelial cells; (ii) Lad is tyrosine phosphorylated upon PDGF stimulation and is associated with PDGF receptor; (iii) upon PDGF stimulation, Grb2 is recruited to Lad in human embryonic lung cells; (iv) overexpression of Lad elevated AP-1 promoter activity by two- to threefold, whereas dominant negative Lad abrogated PDGF-dependent activation of AP-1 promoter. These results provide a novel mechanism of PDGF-dependent signaling, in which Lad acts as an adaptor in a tissue-specific manner, linking PDGF signal to Grb2 and subsequent activation of AP-1. PMID- 11394875 TI - Expression of attractin and its differential enzyme activity in glioma cells. AB - Attractin/mahogany protein was previously shown to be involved in a number of physiological and pathological events, including immune system regulation, body weight control, pigmentation, myelinization, and tumor susceptibility. Human attractin has an enzymatic activity resembling dipeptidyl peptidase IV (DPP-IV). In the central nervous system, attractin has been detected in neurons but not in glial cells up to now. We show the expression of attractin mRNA and protein in glioma cell lines at different degree of transformation. In human U373 and U87 glioma cells (Grades III and IV), membrane-bound attractin displays hydrolytic activity amounting to 5 and 25% of total cellular DPP-IV-like enzyme activity, respectively. Such activity has not been observed in the rat C6 glioma cells (Grade I). Attractin presence in glioma, but not in normal glial cells, together with its differential enzymatic activity, suggests its role in growth properties of tumors of glial cell origin. PMID- 11394876 TI - An optimized PCR-based procedure for production of 13C/15N-labeled DNA. AB - We have substantially improved a procedure that we previously described for producing 13C/15N-labeled DNA (Chen et al., FEBS Lett. 436, 372-376, 1998) to provide an economical and straightforward approach to the preparation of labeled DNA. The conditions for the PCR reactions have been optimized to permit the use of low concentrations of the costly labeled dNTPs (50 microM for each). In addition, a rapid and high-yield purification procedure has been developed that allows us to obtain a high yield of very pure labeled DNA. These modifications to our original procedure permit us to obtain 1.9 mg of an 18 bp DNA oligomer from 20 mg of dNTPs (ca. 10% yield from the starting dNTPs). This is sufficient material for the preparation of 0.4 mM sample in a volume of 400 microl. In summary, this procedure is a cost-effective, time-efficient procedure for the production of labeled DNA for NMR studies. PMID- 11394877 TI - Structure and expression of the human histamine H4-receptor gene. AB - We report the characterization by genomics-based approach of the human H4 receptor gene structure. The H4-receptor gene have been mapped by radiation hybrid experiments (Gene Bridge 4) on chromosome 18q11.2, between the AFMBB11WH5 and CHLC.GATA85D10 markers. The H4-receptor gene spans more than 21 kbp and contains three exons separated by two large introns (>7 kbp). RT-PCR analysis showed that the H4-receptor gene encoded a 3.7 kb mRNA which did not seem to be alternatively spliced within its coding region. The H4-receptor transcripts were found to be highly expressed in peripheral tissues implicated in inflammatory responses such as leukocytes, spleen, lung, and liver. In addition, low expression level of the H4-receptor mRNA was also detected in several human brain regions. Analysis of the 5'-flanking region of the H4-receptor gene did not reveal the existence of canonical TATA or CAAT-box. However, several putative regulatory elements mediating TNFalpha or IL-6-stimulated transcriptional activation were detected. The uteroglobin promoter binding factor, known to mediate anti-inflammatory response of uteroglobin, in the lung, was also found in this region. Thus, the description of the H4-receptor gene promoter region will facilitate the elucidation of its transcriptional control by factors secreted during inflammatory responses. PMID- 11394878 TI - Up-regulation of calcineurin Abeta mRNA in the Alzheimer's disease brain: assessment by cDNA microarray. AB - Recent advances in cDNA microarray technology have made it possible to analyze expression of more than 8000 genes. Using this technology, gene expression in the hippocampus containing neurofibrillary tangle-associated lesions from an Alzheimer's disease (AD) patient was compared with expression in the parietal cortex from the same patient that lacked these lesions. We also compared gene expression using a control brain. The top 20 named genes significantly up regulated or down-regulated only in the AD brain were determined. The most up regulated gene proved to be calcineurin Abeta mRNA (CAbeta). In situ hybridization histochemistry revealed that CAbeta was significantly up-regulated in pyramidal neurons of the hippocampus in the AD brain. RT-PCR analysis revealed that CAbeta was up-regulated in the hippocampus from two out of three AD brains while there were no changes in three control brains. Our study suggests that CAbeta may play a crucial role in the pathophysiological mechanisms in AD. PMID- 11394879 TI - Molecular evidence that human ocular ciliary epithelium expresses components involved in phototransduction. AB - Here we report the expression, in the human ocular ciliary epithelium and in a human nonpigmented (NPE) ciliary epithelial cell line, of genes usually restricted to cone and rod photoreceptor cells of the retina. By RT-PCR and DNA sequencing we identified the expression of rhodopsin and components linked to its deactivation, including rhodopsin kinase, recoverin, and visual arrestin. We also detected the expression of transducin (T-alpha), phosphodiesterase (PDE-alpha), and cGMP-gated channel alpha-subunits. Cultured NPE cells responded to treatment with phorbol ester by enhancing the expression of rhodopsin mRNA three- to fourfold. Indirect immunofluorescence of the intact ciliary epithelium with monoclonal antibodies (MAbs) against rhodopsin, rhodopsin kinase, and visual arrestin revealed labeling preferentially restricted to the NPE cells. Furthermore, Western blot analysis of whole lysates from the pars plicata region of the human ciliary epithelium with MAbs demonstrated immunochemical cross reactivity with proteins of molecular mass similar to rhodopsin (36 kDa), rhodopsin kinase (64 to 66 kDa), and arrestin (48-52 kDa) from the human retina. These results provide the first molecular evidence that components of a non visual phototransduction pathway are expressed in the human ocular NPE ciliary epithelium, which may be linked to circadian entrainment tasks. PMID- 11394880 TI - Cholesterol signaling at the endoplasmic reticulum occurs in npc1(-/-) but not in npc1(-/-), LDLR(-/-) mice. AB - It remains controversial whether deficiency of the Niemann-Pick C1 (npc1) protein results in altered cholesterol signaling at the endoplasmic reticulum (ER). In this report, we have measured the processed, nuclear form of sterol regulatory element binding protein (SREBP)-1 in livers of npc1 wild-type, heterozygous, and homozygous deficient mice, alone, and in combination with deficiencies of the low density lipoprotein receptor (LDLR) or the multiple drug resistant (mdr)1a, P glycoprotein. Cleavage of SREBPs to activated forms normally occurs when the ER is deficient in cholesterol. A large decrease in processed SREBP-1 was evident in fasted npc1(-/-) mice and npc1(-/-), mdr1a(-/-) mice, with no decrease evident in npc1(-/-), LDLR(-/-) mice. These results suggest that the increase in cellular cholesterol which occurs in npc1(-/-) and in npc1(-/-), mdr1a(-/-) mice includes the sites responsible for cholesterol signaling, while the similar increase in cholesterol found in npc1(-/-), LDLR(-/-) mice does not. PMID- 11394881 TI - Colonocyte basolateral membranes contain Escherichia coli heat-stable enterotoxin receptors. AB - Heat-stable enterotoxin (ST(a)) elaborated by E. coli is a major cause of diarrhea. The transmembrane protein guanylyl cyclase C (GC-C) is the acknowledged receptor for ST(a) and for the mammalian peptides guanylin and uroguanylin. Binding to GC-C results in generation of cGMP, activation of type II cGMP dependent protein kinase, phosphorylation of CFTR and increased chloride and bicarbonate secretion. We had previously shown that ST(a) receptors (GC-C) are found on the brush border membranes of small intestinal enterocytes and of colonocytes. However, since it has subsequently been shown that the endogenous ligands for these receptors, guanylin and uroguanylin, circulate in blood, we proposed the existence of ST(a) binding sites on the basolateral membranes (BLM) of colonocytes. Specific binding of 125I-ST(a) to rat colonocyte BLM was seen. The kinetics of binding to the BLM were similar to binding to BBM. The nature of the BLM receptor is unknown. This suggests that circulating guanylin and uroguanylin, analogues of ST(a), may also function via the basolateral surface. PMID- 11394883 TI - Cloning and characterization of a tumor-associated antigen, beta-casein-like protein. AB - Monoclonal antibody (MAb) 1C5 reacts with 87% of uterine cervical adenocarcinoma and bovine beta -casein, but not with squamous cell carcinoma. To clarify the characteristics of the antigen (beta-casein-like protein; BCLP) recognized by MAb 1C5, molecular cloning was performed using 5' rapid amplification of the cDNA ends (5' RACE) and the oligo-capping method. The protein predicted from the cDNA consisting of 937 nucleotides comprises 222 amino acids. The BCLP gene and deduced amino acid sequences were novel and showed no similarity to known cancer associated genes in the database. Northern blot analysis showed that a 1.1 kb transcript was ubiquitously expressed in cancer cell lines and was predominantly expressed in uterine cervical adenocarcinoma. To clarify the function of BCLP, BCLP cDNA was transfected into L929 cells, resulting in a significant increase in cell area, a downregulation of cell growth rate and a decrease in cell attachment. We conclude that BCLP might be associated with cell morphology and a regulation of growth pattern of tumor. PMID- 11394882 TI - Chaperone-mediated regulation of hepatic protein secretion by caloric restriction. AB - Calorie restriction (CR) delays age-related physiological changes, reduces cancer incidence, and increases maximum life span in mammals. Here we show that CR decreased the expression of many hepatic molecular chaperones and concomitantly increased the rate and efficiency of serum protein secretion. Hepatocytes from calorie-restricted mice secreted twice as much albumin, 63% more alpha1 antitrypsin, and 250% more of the 31.5-kDa protein 2 h after their synthesis. A number of trivial explanations for these results, such as differential rates of protein synthesis and cell leakage during the assay, were eliminated. These novel results suggest that CR may promote the secretion of serum proteins, thereby promoting serum protein turnover. This may reduce the circulating level of damaging, glycoxidated serum proteins. PMID- 11394884 TI - Amino-terminal fragment of urokinase-type plasminogen activator inhibits HIV-1 replication. AB - CD8+ T lymphocytes have been shown to produce unidentified soluble factors active in suppressing HIV-1 replication. In this study, we purified an HIV-1 suppressing activity from the culture supernatant of an immortalized CD8+ T cell clone, derived from an HIV-1 infected long-term nonprogressor, and identified this activity as the amino-terminal fragment (ATF) of urokinase-type plasminogen activator (uPA). ATF is catalytically inactive, but suppresses the release of viral particles from the HIV-1 infected cell lines via binding to its receptor CD87. In contrast, cell proliferation and the secretion of an HIV-1 LTR driven reporter gene product were not affected by ATF. These findings suggest that ATF may inhibit the assembly and budding of HIV-1, which provides a novel therapeutic strategy for AIDS. PMID- 11394885 TI - Hypoxia-inducible factor 1alpha and 1beta proteins share common signaling pathways in human prostate cancer cells. AB - Hypoxia-inducible factor 1 (HIF-1) is a heterodimeric transcription factor consisting alpha and beta subunits. It is critically involved in cancer cell hypoxia adaptation, glycolysis, and angiogenesis. HIF-1beta is associated with HIF-1 functions as a dimerization partner of HIF-1alpha, and is on the other hand associated with carcinogenesis via dioxin signaling. Regulation of HIF-1beta protein expression was investigated in human prostate cancer (PCA) cells. HIF 1beta protein was expressed constitutively under nonhypoxic conditions in all human PCA cells tested, and was up-regulated by hypoxia, CoCl2, EGF, serum, or PMA in moderate levels. Compared to that of HIF-1alpha, the constitutive, serum-, EGF-, and PMA-increased HIF-1beta protein expression were also inhibited by selective PI3K or FRAP/TOR inhibitors but in higher doses. Hypoxia partially reversed the dose dependent inhibition of HIF-1beta. These results suggest that HIF-1alpha and beta share common signaling pathways for nuclear protein accumulation. PMID- 11394886 TI - An eukaryotic-type serine/threonine protein kinase involved in the carbon source dependent pigment biosynthesis in Amycolatopsis mediterranei U32. AB - The structural gene, pkmA, was cloned and sequenced from a rifamycin SV-producing Amycolatopsis mediterranei U32 strain. The N-terminal portion of the deduced amino acid sequence of pkmA showed significant similarity to the family of serine/threonine protein kinases. It contains all the structural features which are highly conserved in protein kinases, including the Gly-X-Gly-X-X-Gly motif of ATP binding and the essential amino acids known to be important for the recognition of the correct hydroxyamino acid in serine/threonine protein specific kinases. The protein possesses a region rich in Ala and Pro residues around the middle of pkmA open reading frame, which might be involved in the transmembrane function, as suggested by PhoA fusion protein analysis. The pkmA gene was expressed in Escherichia coli as a glutathione S-transferase (GST) fusion protein, and the protein was found to have the activity of autophosphorylation. A double crossover gene replacement was achieved by inserting an aparmycin resistance gene into pkmA in A. mediterranei chromosomal DNA. The phenotypic analysis of the mutant suggested that pkmA gene is involved in carbon source dependent pigment formation in A. mediterranei U32. PMID- 11394887 TI - The central cannabinoid receptor inactivation suppresses endocrine reproductive functions. AB - The function of central cannabinoid (CB1) receptor was investigated in the regulation of the pituitary-gonad axis in CB1 receptor knockout male mouse. Serum luteinizing hormone (LH) and testosterone (T) levels and basal T secretion in vitro of testes were significantly decreased in mutant (CB1-/-) mice. The receptor agonist, anandamide (ANA), suppressed LH and T secretion in wild type (CB1+/+) mice but had no effect in receptor inactivated animals. The results are the first descriptions indicating the direct action of CB1 receptors on LH and T secretion and the immunohistological demonstration of CB1 receptors in the Leydig cells. The results also indicate that CB1 receptors are responsible for the effects of exogenous cannabinoids on reproductive functions. PMID- 11394888 TI - Hepatic expression of SV40 small-T antigen blocks the in vivo CD95-mediated apoptosis. AB - We have previously demonstrated that CD95-mediated apoptosis of hepatocytes is blocked in a murine model of hepatocarcinogenesis due to the expression of SV40 early sequences encoding the large-T and small-t antigens. In this study, we set out to pinpoint the sequences involved in this apoptosis-resistant phenotype, and tested several mutants of the SV40 early region for their ability to confer protection against CD95-induced apoptosis in transgenic mice. We show that resistance to apoptosis is independent of the transforming character of the mutants and demonstrate that the expression of the small-t antigen alone in transgenic mice is sufficient to confer this resistance. Our data also reveal an increased level of activated Akt kinase in these transgenic mice, and this could account for this hitherto unknown function of the SV40 small-t antigen. PMID- 11394889 TI - Identification of a cis-acting element and a novel trans-acting factor of the glutamine synthetase gene in liver cells. AB - In the mammalian liver the expression of the enzyme glutamine synthetase (GS) is restricted to a small population of hepatocytes. In cells expressing the enzyme up to 3.5% of total cellular protein is GS. In order to identify enhancer elements contributing to this extraordinarily high level of expression we focused on a region roughly 2.5 kbp upstream of the GS promoter. Gel mobility shift assays revealed binding of an unknown protein within the most distal part of this region and reportergene assays demonstrated that roughly 60 bp downstream from position -2503 are indispensable for protein binding and the full effect of the enhancer. In UV cross-link analysis a 38 kDa nuclear protein that binds to the sequence was identified in rat hepatocytes. This nuclear protein, designated as upstream binding factor of the GS gene (UFGS) seems to play an important role in high-level expression of GS in liver. PMID- 11394890 TI - Cellular specificity for the activation of fibroblast growth factor-2 by heparan sulfate proteoglycan. AB - Heparan sulfate proteoglycans (HSPGs) promote cellular proliferation through interaction with FGF-2. To examine the role of cellular specificity of HSPG in FGF-2 function, a recombinant soluble isoform of CD44 (rsCD44v3,8-10) was expressed in various cell types; 293 T fibroblasts, the epithelial carcinoma cell lines A431 and HOTZ, the myelomonocytic cell line THP-1, and the Ig-secreting B lymphoblast IM9. The capacity of the recombinant HSPGs expressed in these cell lines to bind and present FGF-2 to the high-affinity receptor FGFR1 was addressed. This novel approach showed a minor difference in the binding and in the FGF-2 stimulating activity of rsCD44v3,8-10 HSPGs from fibroblasts and epithelial cells. However, FGF-2 binding of rsCD44v3,8-10 from IM9 and THP-1 cells was significantly lower, and stimulation of FGF-2 by rsCD44v3,8-10 from these two cell types could not be detected. We tested the possibility that the differences among cell types were related to the functional profile of endogenous HSPGs. The initial survey of a wider panel of cell types revealed high levels of HSPGs synthesis on the surface of 293 T, epithelial and IM9 cells, but low levels on the surface of other cells of hematopoietic origin. Surprisingly, native HSPGs from fibroblasts and epithelial cell lines promoted FGF-2 biological activity to vastly different extents, and cell surface HSPGs from IM9 cells induced an FGF-2 response. Altogether, the results suggested a role for cell-specific HS modification in addition to synthesis as regulatory mechanisms for the cellular specificity of proteoglycan function. PMID- 11394891 TI - Plasminogen binding is increased with adipocyte differentiation. AB - The purpose of this study was to examine the role of the plasminogen system in the development of adipose tissue. Plasminogen binding capacity was determined in differentiated and undifferentiated cells from adipose tissue of plasminogen deficient mice and 3T3 cells, a well-characterized tissue culture model. In 3T3 cells, plasminogen binding was fivefold higher in differentiated cells compared to the undifferentiated cells. Inhibition of binding by carboxyl-terminal lysine analogs was similar for the differentiated and undifferentiated cells with tranexamic acid > EACA > lysine. The binding of plasminogen was concentration dependent and approaches saturation in the both cell types. The number of plasminogen binding sites was tenfold higher in the differentiated compared to the undifferentiated cells. In isolated mature fat cells and stromal cell cultures from mouse adipose tissue, plasminogen binding was also higher in the differentiated mature fat cells and differentiated stromal cells compared to undifferentiated stromal cells. Plasminogen binding was elevated in the differentiated cells from the Plg-/- mice compared to cells from the WT mice. These results suggest that the plasminogen system plays an important role in adipose tissue development. PMID- 11394892 TI - Neuroendocrine cells along the digestive tract express neuropilin-2. AB - Neuropilin-2 (np-2) is a receptor for semaphorin-3F (sema-3F) and semaphorin-3C (sema-3C). These semaphorins repel tips of growing axons that express np-2. In addition, np-2 functions as a receptor for heparin binding forms of the angiogenic factor vascular endothelial growth factor (VEGF) such as VEGF145 and VEGF165. We report that np-2 is strongly expressed in neuroendocrine cells located all along the human digestive tract. Confocal fluorescent microscopy revealed that np-2 is concentrated in vesicle-like structures located near the nucleus at the basolateral side of these cells. In the colon, the np-2-expressing subpopulation of neuroendocrine cell is almost identical with the serotonin producing subpopulation of neuroendocrine cells. Gastrointestinal carcinoid tumors are digestive tract tumors that develop from neuroendocrine cells. Interestingly, most of the carcinoid tumors derived from the colon and the appendix did not contain np-2-producing cells. However, some carcinoid tumors derived from the small intestine and stomach did express low levels of np-2 in isolated foci of cells. By contrast, strong serotonin and chromogranin-A expression was observed in all of the carcinoid tumors that were examined. These results suggest that loss of np-2 expression may accompany tumor progression in carcinoid tumors. PMID- 11394893 TI - Superoxide potently induces ceramide formation in glomerular endothelial cells. AB - Recent evidence suggests that the sphingolipid-derived second messenger ceramide and oxidative stress are intimately involved in apoptosis induction. Here we report that exposure of microcapillary glomerular endothelial cells to superoxide generating substances, including hypoxanthine/xanthine oxidase and the redox cyclers DMNQ and menadione results in a dose-dependent and delayed increase in the lipid signaling molecule ceramide. Long-term incubation of endothelial cells for 2-30 h with either DMNQ or hypoxanthine/xanthine oxidase leads to a continuous increase in ceramide levels. In contrast, short-term stimulation for 1 min up to 1 h had no effect on ceramide formation. The DMNQ-induced delayed ceramide formation is dose-dependently inhibited by reduced glutathione, whereas oxidized glutathione was without effect. Furthermore, N-acetylcysteine completely blocks DMNQ-induced ceramide formation. All superoxide-generating substances were found to dose-dependently trigger endothelial cell apoptosis. In addition, glutathione and N-acetylcysteine also prevented superoxide-induced apoptosis and implied that ceramide represents an important mediator of superoxide-triggered cell responses like apoptosis. PMID- 11394894 TI - BMP-6 enhances chondrogenesis in a subpopulation of human marrow stromal cells. AB - Marrow stromal cells (MSCs) can differentiate into several mesenchymal lineages. MSCs were recently shown to form cartilage in micromass cultures with serum-free medium containing TGF-beta and dexamethasone. Here we found that addition of BMP 6 increased the weight of the pellets about 10-fold and they stained more extensively for proteoglycans. mRNAs for type II procollagen and type X collagen were detected at 1 week and the levels were increased at 3 weeks. We also compared two subpopulation of cultures of MSCs: Small and rapidly self-renewing cells (RS cells) and the large, more mature and slowly replicating cells (mMSCs). The cartilage pellets prepared from cultures enriched for RS cells were about 2.5 fold larger, stained more extensively for proteoglycans, and had levels of mRNA for type II procollagen that were 1.6-fold higher. Also, RS cells retained more of their chondrogenic potential as the cells were passaged. PMID- 11394895 TI - Calcipotriol inhibits autocrine phosphorylation of EGF receptor in a calcium dependent manner, a possible mechanism for its inhibition of cell proliferation and stimulation of cell differentiation. AB - We report in this study that proliferation inhibition of SCC13 cells by calcipotriol was possibly mediated by its inhibitory effect on autocrine activation of EGF receptor. Based on MTT assay, PCNA staining, DAPI staining, and involucrin immunocytochemical staining, we showed that calcipotriol inhibited cell growth and stimulated differentiation but did not induce apoptosis. Western blot analysis of concanavalin-A-bound fraction demonstrated that calcipotriol specifically dephosphorylated 170- and 66-kDa polypeptides from 8 h posttreatment and complete dephosphorylation was observed at 12 h posttreatment. The 170- and 66-kDa polypeptides were confirmed as EGF receptor and Shc, respectively. Calcipotriol-mediated EGF receptor dephosphorylation required the presence of extracellular calcium. Similar kinetics of the dephosphorylation was also observed in HaCaT cells cultured in medium of high calcium concentration. By BrdU labeling, we also showed calcium dependency of calcipotriol for the inhibition of cell proliferation. Therefore, EGF receptor deactivation by calcipotriol might be a mechanism of action for the inhibition of cell proliferation and the stimulation of differentiation in SCC13 cell and HaCaT cells. PMID- 11394896 TI - Association of the Glu298Asp polymorphism in the endothelial nitric oxide synthase gene with essential hypertension resistant to conventional therapy. AB - Endothelial nitric oxide synthase (eNOS) produces nitric oxide (NO) which, after diffusing into vascular smooth muscle cells, activates guanylate cyclase leading to vasodilatation. A polymorphism (894G to T) in exon 7 of the eNOS gene causes the conversion of Glu to Asp in position 298. The recently described crystal structure of the heme domain of eNOS protein shows that Glu298 is fully solvent accessible and distant from regions integral to enzyme function. Searching for phenotypic expression of eNOS gene variants, we genotyped a group of patients with essential hypertension (H, n = 119) for the Glu298Asp polymorphism and compared them with age- and sex-matched healthy normals (N, n = 85). To specify phenotypic expression further, the hypertensive patients were subdivided into one group that responded well to regular antihypertensive therapy (CH, n = 45) and one group that was resistant to the therapy (RH, n = 74). Patients with BP higher than 140/90 mmHg when on adequate lifestyle modification and triple-combination therapy (including diuretics) were considered resistant. In RH and H groups, a significantly higher frequency of T alleles (P = 0.022 and P = 0.046, respectively) was found compared to normotonics (N). In well-controlled hypertonics, the same tendency was found, but did not reach statistical significance. The Glu298Asp polymorphism may contribute to the complex pathogenesis of essential hypertension and may be a factor in the resistance of these patients to conventional antihypertensive therapy. The presence of this allele may thus be predictive of the patients' therapeutic response. PMID- 11394898 TI - Expression of ID family genes in the synovia from patients with rheumatoid arthritis. AB - Rheumatoid arthritis (RA) is a chronic inflammatory disease characterized by aggressive proliferation of synovial tissue leading to destruction of cartilage and bone. To identify molecules which play a crucial role for the pathogenesis, we compared mRNA expression pattern of RA synovium with that of osteoarthritis (OA), using the differential display. From the panel of differentially expressed genes, ID1 (inhibitor of differentiation 1) was considered to be particularly relevant to the pathogenesis of RA, because Id family genes have been shown to play a role in cell proliferation and angiogenesis. To examine whether the up regulation of these genes is consistently observed in the patients with RA, mRNA levels of ID1 and ID3 in the synovial tissues from 13 patients with RA and 6 patients with OA were semi-quantitatively analyzed by RT-PCR. Mean mRNA levels of ID1 and ID3 were significantly elevated in RA synovia compared with OA by 8.6 fold (P = 0.0044) and 3.3-fold (P = 0.0085), respectively. Immunohistochemistry revealed striking staining of Id1 and Id3 in the endothelial cells, suggesting a possible role of Id in severe angiogenesis observed in RA. The expression of Id family genes in the synovium constitutes a new finding of particular interest. Their functional role as well as their contribution to the genetic susceptibility to RA requires further investigation. PMID- 11394897 TI - Morphological and physiological restorations of hereditary form of dilated cardiomyopathy by somatic gene therapy. AB - TO-2 strain hamsters with dilated cardiomyopathy, gene deletion of delta sarcoglycan (SG) and no expression of alpha-, beta-, gamma-, and delta-SG proteins are useful for developing the potential gene therapy of intractable heart failure. We prepared recombinant adeno-associated virus vector including normal delta-SG gene driven by CMV promoter and intramurally administered in vivo. The transfected myocardium induced robust expression of both transcript and transgene for 2/3 period of the animal's life expectancy. Immunostaining demonstrated reexpression of not only delta-SG but also other three SGs in 40% cells in the transfected region and normalization of the diameter of transduced cardiomyocytes. Hemodynamic study revealed preferential amelioration of the diastolic indices (LVEDP, the dP/dt(min) and CVP). These results provide the first evidence that supplementation of a specific gene with efficient and sustained transfection capability restores the genetic, morphological, and functional deteriorations. PMID- 11394899 TI - cDNA microarray analysis of Helicobacter pylori-mediated alteration of gene expression in gastric cancer cells. AB - Helicobacter pylori infection stimulates several intracellular signaling pathways and is accompanied by increased gene expression in gastric epithelial cells. High density cDNA microarray was used to characterize the mRNA expression profile of genes in human gastric cancer cells (MKN45, AGS) cocultured with H. pylori. Coculture with cag pathogenicity island (PAI)-positive H. pylori (wild-type) significantly up-regulated mRNA expression in 8 of 2304 genes tested. In 6 (interleukin-8, I(kappaB)alpha, A20, ERF-1, keratin K7, glutathione peroxidase) of the 8 genes, up-regulation was confirmed by RT-PCR. In coculture with isogenic cagE-negative mutant ((Delta)cagE), which encodes a type IV secretion system with other genes in the cag PAI, no significant up-regulation was found. We further analyzed the role of A20. Transfection of expression vector encoding A20 resulted in an inhibition of H. pylori-mediated NF-kappaB activation, indicating that H. pylori-mediated A20 expression could be a negative regulator of NF-kappaB activation. Taken together, these results indicate the importance of microarray technology as a tool for analyzing the complex interplay between H. pylori and the host. PMID- 11394900 TI - Transcription factor Yin Yang 1 stimulates poly(ADP-ribosyl)ation and DNA repair. AB - Poly(ADP-ribose) polymerase 1 (PARP-1) is a nuclear enzyme that catalyzes the synthesis of ADP-ribose polymers from NAD(+). The function of PARP-1 is related to important nuclear processes including DNA repair and transcription. Previous studies demonstrated a specific physical interaction between PARP-1 and the transcription factor Yin Yang 1 (YY1) in vitro. In this study, a functional relationship between both proteins in response to genotoxic treatment of cells is presented. The interaction of YY1 with PARP-1 greatly stimulates the enzymatic activity of PARP-1. Consistent with this, the overexpression of YY1 in HeLa cells resulted in an enhanced synthesis of poly(ADP-ribose) and an acceleration of DNA repair in response to a treatment with methyl-N'-nitro-N'-nitrosoguanidine. PMID- 11394901 TI - Identification of a single nucleotide polymorphism in the TATA box of the CYP2A6 gene: impairment of its promoter activity. AB - Human cytochrome P450 2A6 (CYP2A6) constitutes the major nicotine oxidase, and large interindividual differences are seen in the levels of this enzyme, to a great extent caused by the distribution of several different polymorphic gene variants mainly located in the open reading frame (ORF). In the present study, we report a common polymorphism located in the 5' flanking region of CYP2A6 affecting its expression. DHPLC analysis and complete sequence of the open reading frame of the gene from a Turkish individual revealed a -48T > G substitution disrupting the TATA box. Using dynamic allele-specific hybridization (DASH), genotyping of this novel variant (named CYP2A6*9) was carried out in 116 Swedish, 132 Turkish, and 102 Chinese subjects, and the allele frequencies were found to be 5.2, 7.2, and 15.7%, respectively. The significance of the polymorphism was investigated by the construction of luciferase reporter plasmids containing 135 or 500 bp of the 5'-upstream region of the gene transfected into human hepatoma B16A2 cells. The constructs carrying the -48T > G mutation were only expressed at about 50% of the wild-type alleles. It is concluded that the CYP2A6*9 allele might be one of the most common CYP2A6 variants in Caucasians that alters the levels of enzyme expression. PMID- 11394902 TI - Inhibition of volume-regulated anion channels by dominant-negative caveolin-1. AB - Caveolae are flask-shaped invaginations of the plasma membrane formed by the association of caveolin proteins with lipid rafts. In endothelial cells, caveolae function as signal transduction centers controlling NO synthesis and mechanotransduction. We now provide evidence that the endothelial volume regulated anion channel (VRAC) is also under the control of the caveolar system. When calf pulmonary artery endothelial (CPAE) cells were transfected with caveolin-1 Delta1-81 (deletion of amino acids 1 to 81), activation of VRAC by hypotonic cell swelling was strongly impaired. Concomitantly, caveolin-1 Delta1 81 disturbed the formation of caveolin-1 containing lipid rafts as evidenced by sucrose density gradient centrifugation. In nontransfected cells, endogenous caveolin-1 typically associated with low-density, detergent-resistant lipid rafts. However, transient expression of caveolin-1 Delta1-81 caused a redistribution of endogenous caveolin-1 to high-density, detergent-soluble membrane fractions. We therefore conclude that the interaction between caveolin-1 and detergent-resistant lipid rafts is an important prerequisite for endothelial VRAC activity. PMID- 11394903 TI - Soyasaponin I, a potent and specific sialyltransferase inhibitor. AB - A growing number of reports demonstrate that hypersialylation, which is observed in certain pathological processes, such as oncogenic transformation, tumor metastasis, and invasion, is associated with enhanced sialyltransferase (ST) activity. There is therefore a need for the development of ST inhibitors to modulate ST activity and thus alleviate the disease processes caused by STs. In the present study, soyasaponin I had been discovered to be a potent and specific ST inhibitor by screening strategy from 7500 samples including micribial extracts and natural products. Kinetic analysis shows that it is a CMP-Neu5Ac competitive inhibitor with for ST3Gal I with an inhibition constant (K(i)) of 2.1 microM. In addition, it is only active against ST, but not against the other tested glycosyltransferases and glycosidases. Our study is the first report to discover ST inhibitor by screening method and also to provide the new chemical structure information that should be useful in the development of other novel ST inhibitors. PMID- 11394904 TI - Epidermal growth factor stimulation of the ACK1/Dbl pathway in a Cdc42 and Grb2 dependent manner. AB - The tyrosine kinase ACK1 phosphorylates and activates the guanine nucleotide exchange factor Dbl, which in turn directs the Rho family GTP-binding proteins. However, the regulatory mechanism of ACK1/Dbl signaling in response to extracellular stimuli remains poorly understood. Here we describe that epidermal growth factor stimulates the ACK1/Dbl pathway, leading to actin cytoskeletal rearrangements. The role of the two ACK1-binding proteins Cdc42 and Grb2 was assessed by overexpression of the Cdc42/Rac interactive binding domain and a dominant-negative Grb2 mutant, respectively. Specific inhibition of the interaction of ACK1 with Cdc42 or Grb2 by the use of these constructs diminished tyrosine phosphorylation of both ACK1 and Dbl in response to EGF. Therefore, the activation of ACK1 and subsequent downstream signaling require both Cdc42 dependent and Grb2-dependent processes within the cell. In addition, we show that EGF transiently induces formation of the focal complex and stress fibers when ACK1 was ectopically expressed. The induction of these structures was totally sensitive to the action of botulinum toxin C from Clostridium botulinum, suggesting a pivotal role of Rho. These results provide evidence that ACK1 acts as a mediator of EGF signals to Rho family GTP-binding proteins through phosphorylation and activation of GEFs such as Dbl. PMID- 11394905 TI - Compactin enhances osteogenesis in murine embryonic stem cells. AB - Embryonic stem (ES) cells have the capacity to differentiate into various cell types in vitro. In this study, we show that retinoic acid is important for the commitment of ES cells into osteoblasts. Culturing retinoic acid treated ES cells in the presence of the osteogenic supplements ascorbic acid and beta glycerophosphate resulted in the expression of several osteoblast marker genes, osteocalcin, alkaline phosphatase, and osteopontin. However, there was only a slight amount of mineralized matrix secretion. Addition of bone morphogenic protein-2 or compactin, a drug of the statin family of HMG-CoA reductase inhibitors, resulted in a greatly enhanced formation of bone nodules. Compactin did not modify the expression of osteogenic markers, but at the late stage of differentiation promoted an increase in BMP-2 expression. These results establish ES-cell derived osteogenesis as an effective model system to study the molecular mechanisms by which the statin compactin promotes osteoblastic differentiation and bone nodule formation. PMID- 11394906 TI - Glycogen synthase kinase 3beta is tyrosine phosphorylated by PYK2. AB - Glycogen synthase kinase 3beta (GSK3beta) is a Ser/Thr kinase that is involved in numerous cellular activities. GSK3beta is activated by tyrosine phosphorylation. However, very little is known about the tyrosine kinases that are responsible for phosphorylating GSK3beta. In this report, we investigated the ability of the calcium-dependent tyrosine kinase, proline-rich tyrosine kinase 2 (PYK2) to tyrosine phosphorylate GSK3beta. In transfected CHO cells, it was demonstrated that PYK2 tyrosine phosphorylates GSK3beta in situ. The two kinases also coimmunoprecipitated. Furthermore, GSK3beta was tyrosine phosphorylated in vitro by an active, wild type PYK2, but not by the inactive, kinase dead form of PYK2. Therefore, this study is the first to demonstrate that GSK3beta is a substrate of PYK2 both in vitro and in situ. PMID- 11394907 TI - The insulin-sensitive GLUT4 storage compartment is a postendocytic and heterogeneous population recruited by acute exercise. AB - Insulin and acute exercise stimulate glucose transport in skeletal muscle by translocating GLUT4 glucose transporters to the cell surface. GLUT4 is distributed in skeletal muscle in two intracellular membrane populations, an endosomal pool that remains unaltered after insulin treatment and an storage population that is markedly GLUT4 depleted in response to insulin. Here we have further characterized the storage GLUT4 compartment in regard to protein composition and sensitivity to acute exercise. This GLUT4 compartment contained IRAP (insulin-regulated aminopeptidase), transferrin receptors or mannose-6 phosphate/IGF-II receptors, indicating a postendocytic origin. Insulin administration caused a depletion of GLUT4 and IRAP but no changes in transferrin receptors, which suggests that this pool is heterogeneous. In addition, acute exercise caused a marked GLUT4 depletion in the storage compartment, whereas no changes were detected in the endosomal population. In all, our data indicate that the GLUT4 storage population represents a postendocytic and heterogeneous compartment; the storage compartment represents the recruitment site that triggers GLUT4 translocation to the cell surface in response to both insulin and acute exercise. PMID- 11394908 TI - Distinct characteristic of Galpha(h) (transglutaminase II) by compartment: GTPase and transglutaminase activities. AB - Galpha(h) (transglutaminase II) is a bifunctional enzyme possessing transglutaminase and GTPase activities. To better understand the factors affecting these two functions of Galpha(h), we have examined the characteristics of purified Galpha(h) from membrane and cytosol. GTP binding activity of mouse heart Galpha(h) was higher in membrane than that from cytosol. Furthermore, phospholipase C-delta1 (PLC-delta1) activity and coimmunoprecipitation of Galpha(h)-coupled PLC-delta1 in the alpha(1)-adrenoceptor-Galpha(h)-PLC-delta1 complex preparations were increased by phenylephrine in the presence of membranous Galpha(h). On the other hand, transglutaminase activity of cytosolic Galpha(h) was higher than that from membrane Galpha(h). These results demonstrate that bifunctions of Galpha(h) are regulated by its localization that can reflect the cellular functions of Galpha(h). PMID- 11394909 TI - Improved glucose tolerance via enhanced glucose-dependent insulin secretion in dipeptidyl peptidase IV-deficient Fischer rats. AB - Glucagon-like peptide-1 (GLP-1) is an incretin, which induces glucose-dependent insulin secretion. GLP-1 is rapidly degraded by dipeptidyl peptidase IV (DPPIV) after its release. We investigated whether DPPIV-deficient F344/DuCrj rats show improved glucose tolerance when compared with DPPIV-positive F344/Jcl rats. Oral glucose tolerance test indicated improved glucose tolerance in F344/DuCrj rats, but blood glucose levels of the two strains were almost the same 120 min after the glucose bolus. Valine-pyrrolidide, a DPPIV inhibitor, had no effect on the glucose tolerance of F344/DuCrj rats, but improved that of F344/Jcl rats. Enhanced insulin secretion and high plasma active GLP-1 levels were detected in an intraduodenal glucose tolerance test. Glucose tolerance is improved in DPPIV deficient F344/DuCrj rats via enhanced insulin release mediated by high active GLP-1 levels. Our results suggest that DPPIV inhibition is a rational strategy to treat diabetic patients by improving glucose tolerance with low risk of hypoglycemia. PMID- 11394910 TI - Rb-associated protein 46 (RbAp46) inhibits transcriptional transactivation mediated by BRCA1. AB - The retinoblastoma suppressor (Rb)-associated protein 46 (RbAp46) is a member of the WD-repeat protein family and a component of the histone modifying and remodeling complexes. Previously, we demonstrated that RbAp46 is a potent growth inhibitor that can suppress the transformed phenotype of tumor cells. To explore the molecular mechanisms of RbAp46 function, we used RbAp46 as a bait in a yeast two-hybrid screening and found that RbAp46 interacts specifically with the C terminal region of BRCA1 (the BRCT domain), a domain involved in the t transactivation activity of BRCA1. Coimmunoprecipitation assays demonstrated that the interaction of RbAp46 with BRCA1 requires the first two of the four Trp-Asp (WD)-repeats of RbAp46. We also showed that expression of RbAp46 represses the transactivation activity mediated by the BRCT/Gal4 fusion protein and inhibits the transactivation of the p21 promoter mediated by the full-length BRCA1. Interestingly, the association of BRCA1 and RbAp46 is disrupted in cells treated with DNA-damaging agents. These results suggest that RbAp46 may specifically interact with BRCA1 and modulate its transactivation activity in response to DNA damage. PMID- 11394911 TI - Regions of FtsZ important for self-interaction in Staphylococcus aureus. AB - Since FtsZ is structurally similar to eukaryotic tubulin, selective regions of S. aureus FtsZ have been swapped with equivalent regions from tubulin. These mutant FtsZ proteins were analyzed for their interaction with both wildtype FtsZ and FtsA using the yeast two-hybrid assay. Specific regions were identified which abrogated FtsZ-FtsZ binding but unaffected the FtsA-FtsZ interaction. This work will further our understanding of the regions of FtsZ critical for its physiological role. PMID- 11394912 TI - GLUT4 ablation in mice results in redistribution of IRAP to the plasma membrane. AB - Glucose transporter (GLUT) 4 is the insulin responsive glucose transporter in adipose tissue, skeletal muscle, and heart. Insulin elicits increased glucose uptake by recruiting GLUT4 from a specialized intracellular storage site to the cell surface. Expression of various proteins that colocalize with GLUT4 and/or are involved in insulin-stimulated GLUT4 translocation was examined in adipocytes as well as skeletal and cardiac muscles from GLUT4 null mice. Our data demonstrate that expression of insulin-regulated aminopeptidase (IRAP) is divergently regulated in GLUT4 null tissues, e.g., upregulated 1.6-fold in GLUT4 null adipocytes and downregulated in GLUT4 null skeletal muscle (40%) and heart (60%). IRAP exhibited abnormal subcellular distribution and impaired insulin stimulated translocation in GLUT4-deficient tissues. We propose the compartment containing IRAP and proteins normally associated with GLUT4 vesicle traffics constitutively to the cell surface in GLUT4 null adipocytes and skeletal muscle. PMID- 11394913 TI - Ca2+-independent activity of nitric oxide synthase. AB - Ca2+-independent forms of nitric-oxide synthase have significant activity when the endogenous calmodulin subunit is Ca2+ free. Further activation is seen when Ca2+ is added. We have examined the activation of a Ca2+-independent nitric-oxide synthase variant and its two point mutants that are more dependent on Ca2+ for activation using mutant calmodulins containing non-functional Ca2+-binding sites. These studies provide evidence that the Ca2+-independent activity of these enzymes can be exerted through specific adapted interactions between the enzyme and the Ca2+-binding site 2 of calmodulin. Further, the results suggest that EGTA sensitive metals other than Ca2+ complexed to calmodulin may be involved in maximal activation of these nitric-oxide synthase variants. PMID- 11394914 TI - Time-resolved sperm responses to an egg peptide measured by stopped-flow fluorometry. AB - Speract, a decapeptide from sea urchin egg jelly, induces various sperm responses. Stopped-flow fluorometry was used to examine the binding of labeled speract and the intracellular changes in pH (pH(i)) and Ca2+ ([Ca2+]i) it induces in sperm. We observed significant time delays for the increase in pH(i) and [Ca2+]i induced by 200 nM speract (69 and 190 ms, respectively). Also, we found that the receptor undergoes a pH(i)-dependent affinity change at around 129 ms. These time delays probably reflect biochemical processes underlying each sperm response to speract that circumscribe the time sequence of the signaling events. PMID- 11394915 TI - Structural and functional study of reconstituted peripheral benzodiazepine receptor. AB - Recombinant mouse 18 kDa peripheral-type benzodiazepine receptor (PBR) protein was overexpressed in Escherichia coli and isolated using a His. Bind metal chelation resin. Recombinant PBR protein was purified with sodium dodecyl sulfate and reincorporated into liposomes using Bio-Beads SM2 as a detergent removing agent. Negative staining of the reconstituted PBR samples, examined by electron microscopy, showed the formation of proteoliposomes. Freeze-fracture of these proteoliposomes revealed the presence of transmembranous particles of an average size of 3.5 +/- 0.25 nm, consistent with the presence of a monomeric form of the recombinant PBR protein. The reconstituted protein exhibited the ability to bind both the PBR drug ligand isoquinoline carboxamide PK 11195 and cholesterol with nanomolar affinities. These data suggest that a PBR monomer is the minimal functional unit, binding drug ligands and cholesterol. PMID- 11394916 TI - Resuscitation of dormant Mycobacterium tuberculosis by phospholipids or specific peptides. AB - The presence of dormant tubercle bacilli presents a major problem for tuberculosis treatment. The culture supernatant of Mycobacterium tuberculosis was previously shown to resuscitate dormant bacilli in vitro. Here we report identification of active components as phospholipids and a tuberculosis protein Rv1174c. Remarkably, dormant bacilli from a one year old culture which failed to form any colonies could be resuscitated with peptides derived from Rv1174c and formed 10(5-7) colonies/ml. This finding represents the first unambiguous demonstration of resuscitation of dormant tubercle bacilli in vitro and may have implication for the study of mycobacterial dormancy and the design of novel strategies for improved treatment of tuberculosis. PMID- 11394917 TI - Effects of intrathecal morphine, baclofen, clonidine and R-PIA on the acute allodynia-like behaviours after spinal cord ischaemia in rats. AB - The present study assessed the efficacy and potency of intrathecal (i.t.) administration of the opiate morphine, the gamma-aminobutyric acid-B (GABA(B)) receptor agonist baclofen, the alpha2-adrenoceptor agonist clonidine and the adenosine A1-receptor agonist R-phenylisopropyl-adenosine (R-PIA) on the acute allodynia-like behaviour after photochemically induced spinal cord injury (SCI) in rats. Rats displaying allodynia-like behaviours to brushing, von Frey hairs and cold stimulation 1-2 days after photochemically induced SCI were studied. In a cumulative dose regime, morphine (0.1-10 micrcog), baclofen (0.1-1 microg), clonidine (0.1-10 microg) and R-PIA (0.01-10 nmol) were administered i.t. through an implanted catheter at the lumbar spinal cord. All tested drugs dose dependently reduced the brushing, von Frey hairs and cold stimulation-induced allodynia-like behaviour. No increase in adverse effects such as motor deficits was found for morphine, clonidine and R-PIA. There was a slight increase in motor impairments at the highest dose of baclofen. For the mechanical allodynia, morphine appeared to be most effective, whereas baclofen, clonidine and R-PIA only provided a partial alleviation. For the cold allodynia, morphine and baclofen were more effective than clonidine and R-PIA. In relieving acute mechanical and cold allodynia-like behaviours in rats 1-2 days after SCI, i.t. morphine and baclofen were superior to clonidine and R-PIA. PMID- 11394918 TI - Investigating the effects of anxiety sensitivity and coping on the perception of cold pressor pain in healthy women. AB - Research indicates that anxiety sensitivity may be related to the negative experience of pain, especially amongst women. Further evidence with chronic pain patients indicates that anxiety sensitivity may result in avoidance pain-coping strategies. However, this effect has not yet been experimentally investigated in healthy groups. Therefore, the current study sought to investigate the effect of anxiety sensitivity and coping on women's responses to pain. Thirty women who were classified as high in anxiety sensitivity and 30 women classified as low in anxiety sensitivity participated. Within each anxiety sensitivity group, half the participants (n = 15) were randomly instructed to either focus on or avoid cold pressor pain sensations. As expected, women high in anxiety sensitivity were found to report higher levels of sensory and affective pain. Also, and consistent with previous research into anxiety sensitivity, no differences were found between anxiety sensitivity groups for measures of pain threshold or pain tolerance. The pain coping instruction manipulation was found to moderate pain experience, in that the avoidance strategy resulted in higher pain ratings compared to when instructed to focus. Finally, high anxiety sensitive women reported greater pain when instructed to avoid rather than focus on cold pressor pain. These results are discussed in light of previous research and future directions for pain management. PMID- 11394919 TI - Commentary: Anxiety, sensitivity and the pain experience. PMID- 11394920 TI - Long-term fluctuations of pressure pain thresholds in healthy men, normally menstruating women and oral contraceptive users. AB - The aim of this investigation was to evaluate whether the pressure pain threshold (PPT) in masticatory muscles of symptom-free subjects was influenced by fluctuations of the sex hormones. The PPT was measured with an electronic algometer for at least 10 consecutive menstrual cycles in 10 women using oral contraceptives and 10 women not using oral contraceptives, with a regular menstrual cycle (26-31 days). In addition, 10 men were measured in a regular pattern over a period of 1 year. All subjects were symptom-free with an age range between 18 and 39 years. Measurement sessions were held during three different cycle phases (follicular, luteal, perimenstrual) and each session consisted of four consecutive PPT measurements. By means of a linear mixed model (SAS), the PPTs of the masster, temporalis and thumb muscles were compared between: (1) groups, (2) sex-hormonal phases, (3) the four consecutive measurements of each muscle per session and (4) time. The PPTs of the masseter (p = 0.8419) and temporalis muscles (p = 0.2786) did not change significantly over time. There was no significant difference in variance for the masseter (p = 0.6250), temporalis (p = 0.9705) and thumb (p = 0.7446) between the three groups. The PPTs of all muscles were significantly lower during the perimenstrual phases in the two female groups. The present data showed similar patterns of PPTs for the three muscle groups. Moreover, the results have shown a very good consistency of the PPTs over a long time period, both in males and females. PMID- 11394921 TI - Outcome of surgery for cervical radiculopathy evaluated by determination of trapezius muscle microcirculation and electromyography. AB - Surgery for cervical radiculopathy was evaluated in 27 patients after anterior Cloward procedure (19 patients) or posterior decompression (eight patients). In addition, we examined 10 conservatively treated patients. Each patient was studied prospectively with regards to the effects on microcirculation in the local trapezius muscle during a fatiguing series of stepwise increased contractions. The right and left muscles were simultaneously examined pre operatively and postoperatively after 9 months using laser-Doppler flowmetry and simultaneous surface electromyography (EMG). Preoperatively, a reduced microcirculation was found in the most painful side compared with the opposite side. This is in accordance with earlier reports on patients with chronic neurogenic neck pain, who also show reduced muscle tension on EMG. Postoperatively, the muscle blood flow became increased, but only in patients operated on via a posterior approach. A tendency at increased EMG-amplitude and reduced mean power frequency of the EMG was noted. These EMG signs of muscle fatigue suggest increased ability to exhaust the trapezius muscle postoperatively. The observed postoperative changes were consistently more frequent in the less painful side. We conclude from these objective measurements showing only a tendency at increased microcirculation and muscle tension postoperatively, that the effect on the trapezius muscle is limited. PMID- 11394922 TI - Nociceptor activation and protein extravasation induced by inflammatory mediators in human skin. AB - Protein extravasation (PE) is known to play an important role in inflammatory conditions. In this study we used dermal microdialysis to apply inflammatory mediators (histamine, bradykinin, serotonin) to human skin. Locally induced PE was compared to pain ratings and axon reflex erythema measured simultaneously. Linear microdialysis capillaries (outer diameter 0.4 mm; cut-off 3000 kDa) were inserted intracutaneously at a length of 1.5 cm in the volar forearm of healthy volunteers. The capillaries were perfused with Ringer's solution at a constant flow rate of 4 microl/min. The perfusate was sampled at 15-min intervals and was analysed for total protein concentration. After a baseline of 60 min, the perfusion was switched to inflammatory mediators for 30 min and then back to vehicle again. Sensations evoked by the stimulation were assessed on a visual analogue scale and visible axon reflex erythema was measured planimetrically.Dose dependent increases in PE could be assessed for all inflammatory mediators tested. Bradykinin (10(-7)M) induced a significant PE, whereas serotonin was effective only at a concentration of 10(-3)M. While serotonin in lower concentrations induced moderate burning pain and an axon reflex flare but no PE, bradykinin provoked PE without pain or axon reflex flare at a concentration of 10(-7)M. Application of histamine similarly evoked PE at lower concentrations as compared to the induction of itch sensation and axon reflex flare. It is concluded that there is no link between nociceptor activation and protein extravasation induced by inflammatory mediators in healthy human skin. PMID- 11394923 TI - The effect of sympathetic activity on thermal hyperalgesia in capsaicin-treated skin during body cooling and warming. AB - An adrenergic mechanism is thought to contribute to pain in conditions that sometimes develop during chronic inflammation and after nerve or tissue injury. There is some doubt, however, about whether adrenergic activity influences nociception in acute inflammation. To investigate this issue, the noncompetitive alpha-(alpha) adrenergic antagonist phenoxybenzamine was introduced by iontophoresis into the skin of 16 healthy volunteers either before or after the topical application of capsaicin. When applied before capsaicin, phenoxybenzamine increased thermal hyperalgesia at normal ambient temperatures and during body warming. These findings suggest that phenoxybenzamine blocked an analgesic mechanism when applied before the onset of inflammation. However, this effect disappeared during body cooling. When applied after capsaicin, phenoxybenzamine inhibited thermal hyperalgesia at normal ambient temperatures, and during body warming and cooling. Thus, phenoxybenzamine blocked a hyperalgesic mechanism when applied after the onset of inflammation. It was concluded that the presence of inflammation influences the nociceptive effect of alpha-adrenergic blockage, possibly by increasing access to excitatory adrenergic receptors on nociceptive afferents. An excitatory adrenergic influence on nociception may overcome an inhibitory adrenergic influence during inflammation. PMID- 11394924 TI - The neurokinin-1 receptor antagonist RP 67580 reduces the sensitization of primary afferents by substance P in the rat. AB - The inflammatory mediator substance P (SP) produces a variety of biological effects in several tissues by binding to the tachykinin receptor neurokinin 1 (NK1) and, to a lesser extent, by binding to the neurokinin 2 receptor (NK2). To assess the sensitizing effect of SP on articular afferent fibres the NK1receptor antagonist RP 67580 was applied in normal and acutely inflamed rat knee joints. Altogether 38 fine afferent nerve fibres from the rat knee with conduction velocities of 0.71-13.5 m/s were recorded as single units, during non-noxious and noxious joint rotations. SP, injected i.a. as a bolus close to the knee joint, was able to sensitize 45.5% (10 of 22) of the units recorded from normal joints and 33.3% (five of 15) of afferents from inflamed joints. The following i.a. application of RP 67580 in a range of 20-200 nmol antagonized in a dose-dependent manner the sensitizing effect of SP in a large proportion of slowly conducting articular afferents from normal (66.7%) and inflamed (46.2%) knee joints. Subsequent SP application enhanced the afferent sensitivity further. The electrophysiological results presented here further support the suggestion that the sensitization of afferents by SP in the rat knee joint is mediated mainly by the NK1 receptor, which is probably located on the primary afferents. PMID- 11394925 TI - Knee joint mobilization reduces secondary mechanical hyperalgesia induced by capsaicin injection into the ankle joint. AB - Joint mobilization is a treatment approach commonly used by physical therapists for the management of a variety of painful conditions. However, the clinical effectiveness when compared to placebo and the neurophysiological mechanism of action are not known. The purpose of this study was to establish that application of a manual therapy technique will produce antihyperalgesia in an animal model of joint inflammation and that the antihyperalgesia produced by joint mobilization depends on the time of treatment application. Capsaicin (0.2%, 50 microl) was injected into the lateral aspect of the left ankle joint and mechanical withdrawal threshold assessed before and after capsaicin injection in Sprague Dawley rats. Joint mobilization of the ipsilateral knee joint was performed 2 h after capsaicin injection for a total of 3 min, 9 min or 15 min under halothane anaesthesia. Control groups included animals that received halothane for the same time as the group that received joint mobilization and those whose limbs were held for the same duration as the mobilization (no halothane). Capsaicin resulted in a decreased mechanical withdrawal threshold by 2 h after injection that was maintained through 4 h. Both 9 and 15 min of mobilization, but not 3 min of mobilization, increased the withdrawal threshold to mechanical stimuli to baseline values when compared with control groups. The antihyperalgesic effect of joint mobilization lasted 30 min. Thus, joint mobilization (9 or 15 min duration) produces a significant reversal of secondary mechanical hyperalgesia induced by intra-articular injection of capsaicin. PMID- 11394926 TI - The morbidity, time course and predictive factors for persistent post-thoracotomy pain. AB - After thoracotomy, patients often suffer from a persistent pain syndrome called post-thoracotomy pain. To elucidate morbidity, time course, and predictive factors for this syndrome, we analyzed follow-up data for 85 post-thoracotomy patients. We used a four-point scale to assess pain: none, slight, moderate and severe. Of 85 patients, 50 reported pain (39 slight, 11 moderate) one day after surgery. A year after surgery, the patients were polled using a simple questionnaire received by the mail. Sixty patients reported persistent pain (34 slight, 14 moderate, 12 severe) a month after surgery, and 35 patients reported persistent pain (33 slight, two moderate) around the time of the poll (1 year after surgery). Although pain deterioration was observed in 40% (34/85) of patients during month 1 after surgery, pain alleviation was seen in 48% (41/85) of patients during months 2-12. Stepwise regression analysis revealed that female gender and pain at postoperative day 1 were predictive for persistent pain both 1 month and 1 year after thoracotomy. Among 35 patients with persistent pain 1 year after surgery, 24 cases reported paresthesia-dysesthesia, and 14 cases reported hypoesthesia. The present data thus suggests that persistent pain is common and often severe 1 month after surgery but is alleviated after 1 year. Clinical time course and symptoms indicate that nerve impairment rather than simple nociceptive impact may be involved in this syndrome. PMID- 11394927 TI - Interactions of histamine and bradykinin on polymodal C-fibres in isolated rat skin. AB - Patients suffering from pruritus due to atopic dermatitis show, in asymptomatic skin, reduced itch and flare responses to histamine, the major pruritogenic mediator. We hypothesized that this apparent loss in histamine sensitivity may be overcompensated in inflamed skin and investigated the interactions between histamine and bradykinin, the major inflammatory mediator. The studies were performed using the isolated rat skin-nerve preparation. Forty-two fibres were tested following four different experimental protocols. After characterization of the sensory properties, six fibres were treated repetitively with histamine (HIS1, HIS2) to exclude the possibility that the responses (spikes/min) increase simply by repetition. In 12 other units, histamine (HIS1) was followed by a wash out period prior to bradykinin (BK) stimulation; in another 12 units, BK followed immediately after HIS1. A further 12 fibres were examined without preceding heat stimulation in order to avoid possible sensitization. If BK was administered after a wash-out period following HIS1, the BK responses were significantly higher than the HIS1 response. The BK response showed a peak discharge which was absent if BK followed directly upon HIS1. If HIS2 was applied directly following BK, the induced discharge was significantly larger than the first histamine response and not different from the BK response, whereas a washout period before HIS2 abolished the sensitizing effect of previous BK.A unidirectional sensitization by previous bradykinin or heat stimulation on the histamine responsiveness of polymodal nociceptors has been demonstrated. If 'itch fibres' in humans were subject to similar interactions of histamine with inflammatory mediators, this may compensate for a down-regulation of histamine receptors in eczematic skin and possibly account for the pruritus. PMID- 11394928 TI - Mediated imitation in 6-month-olds: remembering by association. AB - In two experiments with 72 6-month-olds, we examined whether associating an imitation task with an operant task affects infants' memory for either task. In Experiment 1, infants who imitated target actions that were modeled for 60 s on a hand puppet remembered them for only 1 day. We hypothesized that if infants associated the puppet imitation task with a longer-remembered operant task, then they might remember it longer too. In Experiment 2, infants learned to press a lever to activate a miniature train-a task 6-month-olds remember for 2 weeks-and saw the target actions modeled immediately afterward. These infants successfully imitated for up to 2 weeks, but only if the train memory was retrieved first. A follow-up experiment revealed that the learned association was bidirectional. This is the first demonstration of mediated imitation in 6-month-olds across two very different paradigms and reveals that associations are an important means of protracting memories. PMID- 11394929 TI - Increasing specificity in perceptual development: infants' detection of nested levels of multimodal stimulation. AB - This research assessed the development of infants' sensitivity to two nested amodal temporal relations in audible and visible events. Their detection of global temporal synchrony between visible and audible impacts and internal temporal structure nested within each impact specifying object composition (single versus compound objects) was assessed. Infants of 4, 7, and 11 weeks of age were habituated to a single and a compound object striking a surface and then received test trials depicting a change in synchrony or object composition. Results indicated an interaction between age and condition where sensitivity to synchrony was present by 4 weeks and remained stable across age, whereas sensitivity to composition emerged later, by 7 weeks, and increased dramatically with age. These findings converge with other recent findings to illustrate a pattern of increasing specificity in the development of perception, where infants first detect global and later detect embedded relations. The early sensitivity to global relations may provide an organizational framework for development by focusing infant attention on unitary events, guiding and constraining further exploration, and buffering infants from learning incongruent relations. PMID- 11394930 TI - Long-term retention in 3.5-month-olds: familiarization time and individual differences in attentional style. AB - Using a paired-comparison procedure, we examined the effect of familiarization variables on 3.5-month-old infants' (n = 120) retention of dynamic visual stimuli after 1-min, 1-day, and 1-month delays. The proportion of total looking time to the novel stimulus revealed novelty, null, and familiarity preferences after 1 min, 1-day, and 1-month delays, respectively, for infants who were permitted 30 s of familiarization time. Twenty seconds of familiarization time was insufficient to produce novelty preferences. These results support models of infant retention in which the direction of attentional preferences (novel, familiar, or null) depends on memory accessibility. To examine the impact of individual differences in familiarization or attentional style on memory, infants were identified as long or short lookers according to their peak-look duration on pretest and familiarization trial measures. Compared to long lookers, short lookers showed better retention over time indicating that much of the variability in the infant group data could be accounted for by these individual differences. PMID- 11394931 TI - Mathematical problem solving and working memory in children with learning disabilities: both executive and phonological processes are important. AB - The purpose of this investigation was to explore the relationship between working memory (WM) and mathematical problem solving in children with learning disabilities (LD). Children with LD (age 11.5 years) were compared to chronologically age-matched (CA-M) and younger comprehension/computation achievement-matched children (age 8.9 years) on measures of verbal and visual spatial WM, phonological processing, components of problem solving, and word problem solving accuracy. The results showed that (1) children with LD were inferior on measures of word solution accuracy, components of problem solving, phonological processing, domain-general WM, and verbal WM when compared to children who were CA-M, (2) children with LD were comparable to younger children on all processing measures, except measures of domain-general WM, visual-spatial WM, phonemic deletion, and identifying problem goals, (3) measures of verbal and visual-spatial WM contributed significant variance to solution accuracy independent of phonological processing, and (4) the influence of WM on solution accuracy was mediated by long-term memory (LTM) processes related to the knowledge of algorithms. The results support the notion that information activated from LTM, rather than phonological processing, mediates the relationship between executive processing and solution accuracy in children with LD. PMID- 11394932 TI - Current perspectives on the development of non-biogenic amine-based antidepressants. AB - Compounds that inhibit the re-uptake and/or metabolism of biogenic amines (i.e. serotonin, norepinephrine, and dopamine) have been used to treat depression for more than 40 years. Selective re-uptake inhibitors, currently the most widely prescribed class of biogenic amine-based agents, are certainly safe and relatively easy to use, but do not exhibit either a faster onset of action or greater efficacy than their predecessors. An approach to overcome the limitations that may be inherent to these 'conventional' therapies is to circumvent the monoaminergic synapse. In this review, two potential antidepressant strategies are discussed that may converge with intracellular pathways impacted by chronic treatment with biogenic amine-based agents. Drugs emerging from these strategies may offer significant advantages over currently used antidepressants. PMID- 11394933 TI - Monitoring of L-dopa concentrations in Parkinson's disease. AB - Although it is the treatment of choice in the management of idiopathic Parkinson's disease, l-DOPA (LD) shows a decline in its efficacy after years of prolonged use, with the appearance of severe motor disturbances. These complications have been interpreted on pharmacokinetic and pharmacodynamic grounds. The main pharmacokinetic reason is considered to be the decreased capacity in LD activation to dopamine along with reduction of its storage ability in the nigrostriatal terminals, as a result of disease progression. At this stage the LD action in the extrapyramidal system is thought to be closely dependent on its synaptic cleft concentrations, being directly related to those in the systemic circulation and to the events possibly perturbing them. Therapeutic drug monitoring might be useful to explain these modifications in relation to the clinical effect and even to possible problems in transport competition and so to define the LD dosage regimen. Recently LD threshold concentrations have been suggested by means of sophisticated pharmacokinetic-pharmacodynamic approaches. Unfortunately they do not correspond to real therapeutic ranges but to levels in a hypothetical effect compartment in no steady-state conditions, due to remarkable LD fluctuations. However they are considered helpful to the functional state interpretation of the nigrostriatal system. PMID- 11394934 TI - Anti-inflammatory drugs: new multitarget compounds to face an old problem. The dual inhibition concept. AB - In this short review we have tried to focus on some new relevant aspects of the pharmacological control of inflammation. The clinical availability of new drugs able to produce a selective inhibition of type 2 cyclooxygenase (COX-2), the enzyme thought to be mainly responsible for generating arachidonic-acid-derived inflammatory mediators, has been the origin of much hope. However, expectations of having an effective and completely safe non-steroidal anti-inflammatory drug (NSAID) have been only partially fulfilled. Emerging information has challenged some aspects of the original hypothesis indicating COX-2 as devoid of 'housekeeping' physiological functions. Moreover, the recently available clinical studies have indicated only a relatively small improvement in the tolerability of the newer 'selective' COX-2 inhibitors over the classical COX-1/COX-2 mixed type NSAIDs. The new appreciation of the role of other arachidonic acid derivatives, the leukotrienes (LTS), in producing and maintaining inflammation has generated considerable interest in drugs able to block LTS receptors or to produce a selective inhibition of 5-lipoxygenase (5-LO), the initial key enzyme of the leukotriene pathway. These drugs are now included among the effective therapies of asthma but appear, in the few clinical studies performed, to be an insufficient single therapeutic approach in other inflammatory diseases. Drugs able to block equally well both COX and 5-LO metabolic pathways (dual inhibitors) have been developed and experimentally evaluated in the last few years, but none are available on the market yet. The pharmacological rationale at the basis of their development is strong, and animal studies are indicative of a wide range of anti-inflammatory activity. What appears most impressive from the available studies on dual inhibitors is their almost complete lack of gastric toxicity, the most troublesome side effect of NSAIDs. The mechanism of the gastric-sparing properties of these drugs is not yet completely understood; however, it appears that leukotrienes significantly contribute to gastric epithelial injury particularly when these compounds represent the major arachidonic acid derivatives present in the gastric mucosa after inhibiton of prostanoid production. PMID- 11394935 TI - How to improve the readability of the patient package leaflet: a survey on the use of colour, print size and layout. AB - This paper displays the results of the second part of a survey about patient information and the use of the patient package leaflet. The aim of this research is to investigate the consumers' attitude towards written information. As the formal aspects of the written message are very important in communication, we prepared a questionnaire in order to evaluate the attitude of patients towards some typographical modifications. Patients were invited to give indications about which colours could be used in the different paragraphs of the package leaflet and which print size could be easily read. All people interviewed were asked to choose a colour, from six proposed by us, to be used for 'therapeutic indications', 'side effects', 'how to use', 'paediatric use', 'contraindications', 'use in pregnancy' and 'warnings'. Clear suggestions for the choice of colours for therapeutic indications, side effects and contraindications arose from the survey. In the other cases there was no uniformity of answers. All people complained that the print size used in the package leaflet is too small and suggested 10 and 11 points Didot. Finally, from the survey it emerged that people would appreciate a more detailed package leaflet but information should be given in a schematic and concise way. PMID- 11394936 TI - 1,5-Benzodiazepine tricyclic derivatives exerting anti-inflammatory effects in mice by inhibiting interleukin-6 and prostaglandinE(2)production. AB - The 1,4- and the 1,5-benzodiazepines (BDZ) are commonly used as anxiolytic and anticonvulsive drugs. It has been suggested that they influence, particularly through stimulation of peripheral BDZ receptors, some immune cell properties such as pro-inflammatory cytokine production. The availability of a new class of [1,2,4]triazolo[4,3-a][1,5]benzodiazepine derivatives (compounds IV), endowed with anti-inflammatory and/or analgesic properties but no anti-pentylenetetrazole activity, prompted us to investigate in more detail the anti-inflammatory properties of three selected compounds IV (N,N-dimethyl-1-phenyl-4H [1,2,4]triazolo[4,3-a][1,5]benz- odiazepin-5-amine; N,N-dibutyl-4H [1,2,4]triazolo[4,3-a][1,5]benzodiazepin-5-amine; 1-methyl-N,N-dimethyl-4H [1,2,4]triazolo[4,3-a][1,5]benzodiazepin-5-amine) and one structurally related compound (1-phenyl-4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepin-5(6H)-one). These BDZ derivatives have lost their affinity for the central and peripheral BDZ receptors. The in vivo effect on leukocyte migration of these compounds was investigated by using the mouse air-pouch model of local inflammation. Compounds A and B, significantly inhibited the carrageenan-induced leukocyte recruitment in a dose-dependent manner starting from the dose of 50 mgkg(-1), whereas compound C was effective only at the higher dose of 100 mgkg(-1). Compound D did not exert such effects at any of the doses considered. The effect of compounds A, B and C on leukocyte recruitment was paralleled by a significant inhibition of interleukin-6 and prostaglandin E(2)production in the exudate, similarly to indomethacin, and by a partial reduction of vascular permeability. These features may be relevant for the design and development of innovative anti-inflammatory molecules among the 4H-[1,2,4]triazolo[4,3-a][1,5]benzodiazepin-5-amine derivatives. PMID- 11394937 TI - Platelet inhibition by sertraline and N-desmethylsertraline: a possible missing link between depression, coronary events, and mortality benefits of selective serotonin reuptake inhibitors. AB - Recently, clinical depression has been identified as an independent risk factor for increased mortality in patients following acute coronary events. Although the underlying mechanisms of this link remain uncertain, increased platelet activity has been suggested but never proven as the mechanism responsible for this association. Sertraline hydrochloride is a selective serotonin reuptake inhibitor (SSRI), and is an effective antidepressant agent. Its major liver metabolite, N desmethylsertraline (NDMS), is known to be neurologically inactive. We assessed the in vitro effects of escalating concentrations of sertraline and NDMS on human platelets by aggregometry in plasma and whole blood, by expression of major surface receptors with flow cytometry in washed cells and in the whole blood, and quantitatively by various platelet function analysers in healthy volunteers and patients with coronary artery disease. Pretreatment of blood samples with sertraline and NDMS resulted in a dose-dependent inhibition of platelet-rich plasma aggregation induced by 5 microM ADP (P =, 0.002), by 10 microM ADP (P = 0.0017), by collagen (P = 0.008), and by thrombin (P = 0.026). Whole blood platelet aggregability was also significantly reduced when induced by 20 microM ADP (P = 0.006), and by collagen (P = 0.01). Surface expression of CD9 (P = 0.004), GP Ib (P = 0.0001), GP IIb/IIIa (P = 0.007), VLA-2 (P = 0.01), P-selectin (P = 0.02), and PECAM-1 (P = 0.01), but not the vitronectin receptor, was also reduced in sertraline and NDMS pretreated washed platelets. Whole blood flow cytometry revealed significant inhibition of GP IIb/IIIa (P = 0.008), and P selectin expression (P = 0.0001) in NDMS treated samples. Closure time was delayed for the collagen-ADP cartridge (P = 0.009), and for the collagen epinephrin cartridge (P = 0.01), indicating platelet inhibition in whole blood under high shear conditions. Rapid platelet-function assay revealed a decreased (P = 0.002) ability of platelets to agglutinate fibrinogen-coated beads, suggesting GP IIb/IIIa inhibition. Both sertraline, and its neurologically inactive metabolite NDMS, exhibited significant dose-dependent inhibition of human platelets. The documented anti-platelet effects of sertraline and NDMS may be directly related to the mortality benefits of SSRIs after ischemic events including myocardial infarction and stroke. PMID- 11394938 TI - Protective role of nitric oxide in indomethacin-induced gastric ulceration by a mechanism independent of gastric acid secretion. AB - Gastric ulceration was induced in rats by i.p. injection of the non-steroidal anti-inflammatory drug (NSAID), indomethacin (IND) (30 mg kg(-1)). Pyloric ligation was carried out in each animal before injection to enable collection of the gastric juice. Three hours later, the animals were killed and their stomachs were removed. In the gastric juice, the amounts of mucin, pepsin and HCl were assessed. Gastric mucosa were scrapped for the determination of nitric oxide (NO) (as nitrite) after evaluation of the gastric ulcer index. The influence of arginine (ARG) (300 mg kg(-1)), a NO precursor, N(G)-nitro- l -arginine methyl ester (l -NAME) (50 mg kg(-1)), a non-selective constitutive nitric oxide synthase/inducible nitric oxide synthase (cNOS/iNOS) inhibitor, and the selective iNOS inhibitor aminoguanidine (AMG) (50 mg kg(-1)) were studied. Each NO modulator was injected i.p. 30 min before IND administration. Results indicated that IND elevated gastric acidity by 80% of the normal group, decreased non significantly mucosal nitrite by 22% and exhibited a remarkably high ulcer index (chi = 17). Neither mucin nor pepsin levels were significantly altered. In comparison with the IND group, pretreatment with l -NAME caused a significant decrease in gastric HCl, further decrease in mucosal nitrite (50% of normal) and a two-fold increase in the ulcer index score (chi = 34), despite the decrease in HCl. AMG did not alter gastric acidity, decreased mucosal nitrite by 38% of the normal value and failed to alter significantly the ulcer index of IND. On the other hand, pretreatment with ARG did not alter the gastric acidity and raised mucosal nitrite by 10% above normal. Surprisingly, ARG improved the gastric ulcer score (chi = 1) almost similar to the normal score (chi = zero). Therefore, this study creates a new pathway for the potential treatment of NSAID gastric ulceration through modulation of NO synthesis, regardless of the effect on gastric acidity. PMID- 11394939 TI - Sex-dependent changes in blood-brain barrier permeability in epileptic rats following acute hyperosmotic exposure. AB - Osmotic and ionic balance in the central nervous system is especially regulated by solutes and water transport across the blood-brain barrier. The aim of this study was to investigate the effect of systemic hyperosmolarity on the blood brain barrier permeability in both sexes after experimentally induced seizures. Eight groups of rats were studied: Group I: female control; Group II: male control; Group III: hyperosmotic female; Group IV: hyperosmotic male; Group V: non-hyperosmotic female + seizure; Group VI: non-hyperosmotic male + seizure; Group VII: hyperosmotic female + seizure; Group VIII: hyperosmotic male + seizure. In female rats with pentylenetetrazol-induced seizures, the extravasation of Evans-blue was greater in the brains of hyperosmotic animals than that in normal rats (P < 0.02). However, in male rats, the extravasation of Evans-blue was similar in the brains of hyperosmotic male rats and normal rats after seizure. Our results concluded that hyperosmotic female rats were shown to have a large increase in blood-brain barrier permeability in comparison to hyperosmotic male rats after pentylenetetrazol-induced seizures. PMID- 11394940 TI - Changes of [3H]MK-801, [3H]muscimol and [3H]flunitrazepam binding in rat brain by the prolonged ventricular infusion of ginsenoside Rc and Rg1. AB - In the present study, we have investigated the effects of centrally administered ginsenoside Rc and Rg1 on the modulation of the NMDA receptor and GABA(A)receptor binding in rat brain. The NMDA receptor binding was analysed by quantitative autoradiography using [(3)H]MK-801 binding, and the GABA(A)receptor bindings were analysed by using [(3)H]muscimol and [(3)H]flunitrazepam binding in rat brain slices. Rats were infused with ginsenoside Rc or Rg1 (10 microg/10 microl h(-1), i.c.v.) for 7 days, through pre-implanted cannula using osmotic minipumps (Alzet, model 2ML). The levels of [(3)H]MK-801 binding were highly decreased in part of the parietal layers of the cortex and cingulated by ginsenoside Rc and Rg1. The levels of [(3)H]muscimol binding were strongly elevated in almost all regions of the frontal cortex by the treatment of ginsenoside Rc but decreased by ginsenoside Rg1. However, the [(3)H]flunitrazepam binding was not modulated by ginsenoside Rc or Rg1 infusion. These results suggest that prolonged infusion of ginsenosides could differentially modulate [(3)H]MK-801 and [(3)H]muscimol binding in a region-specific manner. PMID- 11394941 TI - Protective effects of Saiko-ka-ryukotsu-borei-to (Chai-Hu-Jia-Long-Gu-Mu-Li-Tang) against atherosclerosis in Kurosawa and Kusanagi-hypercholesterolemic (KHC) rabbits. AB - We investigated the protective effects of the traditional Japanese herbal medicine Saiko-ka-ryukotsu-borei-to (Chai-Hu-Jia-Long-Gu-Mu-Li-Tang in Chinese) (SRBT) against hypercholesterolemia and atherosclerotic lesions. We focused on atherosclerosis using female heterozygous Kurosawa and Kusanagi hypercholesterolemic (KHC) rabbits. The total plasma cholesterol levels increased for up to 12 weeks after beginning a diet containing 0.1% cholesterol and then reached a plateau of about 600 mg dl(-1). When SRBT was administered at a dose of 1.0 g kg(-1)per day for 24 weeks, total plasma cholesterol levels were significantly decreased after 20-24 weeks. On the other hand, pravastatin at a dose of 10 mg kg(-1)per day produced a significant decrease in total plasma cholesterol levels from 4 to 24 weeks (about 105-130 mg dl(-1)). Moreover, 1.0 g kg(-1)per day of SRBT significantly decreased plasma low density lipoprotein (LDL) cholesterol levels but did not change either very low density lipoprotein (VLDL), or high density lipoprotein (HDL) cholesterol levels. Animals that received pravastatin had significantly decreased LDL cholesterol levels and VLDL cholesterol levels after 8 weeks and at 24 weeks. We also examined the expression of apoB, E and LDL receptor mRNA levels in the liver at 24 weeks after beginning the administration of 1.0 g kg(-1)per day of SRBT. Both apoE and LDL receptor mRNA levels were significantly increased compared with those in rabbits receiving the 0.1% cholesterol diet. SRBT at a dose of 1.0 g kg(-1)per day significantly depressed the intimal surface area of the thoracic aortae involved with atheromatous plaques. The present results suggest that SRBT may protect against hypercholesterolemia and atheromatous lesions by affecting apoE and LDL receptor mRNA gene expression in the liver. PMID- 11394942 TI - Rebamipide prevents periarterial blood-induced vasospasm in the rat femoral artery model. AB - We examined the effect of rebamipide on the vasospasm induced by periarterial blood. In the in vitro study, the significant production of superoxide anion that was identified 3 hours after application of 10% whole blood to the rat aortic segments was inhibited by rebamipide (100 and 300 microM) and these results were correlated with the in vitro intercellular adhesion molecule-1 (ICAM-1) expression on the femoral artery. In the in vivo study, there was an increased mobilization of granulocytes in parallel with a large expression of ICAM-1 in the vessels 24 hours after periarterial blood was applied to the femoral artery which then declined. Subsequently, infiltration of macrophages progressively increased at all layers 7-12 days after application. Pretreatment with rebamipide (100 and 300 mg kg(-1) day(-1), p.o.) significantly inhibited the morphological changes as well as the expression of ICAM-1 with inhibition of granulocyte/macrophage mobilization. In association with these findings, increased wall thickness and decreased lumen area were significantly inhibited by pretreatment with rebamipide. These results provide valuable information for the therapeutic use of rebamipide to relieve vascular remodeling induced by periarterial blood. PMID- 11394943 TI - Transmucosal transport of tobramycin incorporated in solid lipid nanoparticles (SLN) after duodenal administration to rats. Part II--tissue distribution. AB - Tobramycin-loaded solid lipid nanoparticles (SLN) were prepared and administered by duodenal and intravenous (i.v.) routes to rats and the tissue distributions were determined successively at fixed times (30 min, 4 h and 24 h) and compared to those of the tobramycin solution after i.v. administration. The tissue distribution between tobramycin-loaded SLN administered duodenally and i.v. was different. A marked difference between tobramycin-loaded SLN administered duodenally and tobramycin solution administered i.v. was also evidenced. In particular, the amounts of tobramycin in the kidneys after tobramycin-loaded SLN administration either duodenally or i.v. were lower than after administration of i.v. solution. Tobramycin-loaded SLN were able to pass across the blood-brain barrier in rats to a greater extent after i.v. injection than after duodenal administration. PMID- 11394944 TI - Effect of fluoxetine on intracellular Ca2+ levels in bladder female transitional carcinoma (BFTC) cells. AB - The effect of the antidepressant fluoxetine on Ca2+ signaling in cultured cells was largely unknown. The effect of various concentrations of fluoxetine on [Ca 2+] i in populations of bladder female transitional cancer (BFTC) cells was evaluated by using fura-2 as a Ca2+ probe. Fluoxetine increased [Ca 2+] i concentration dependently (20-100 microM) with an EC50 value of 30 microM. The response was inhibited by 50-60% on extracellular Ca2+ removal. In Ca2+ -free medium, pretreatment with 1 microM thapsigargin (an inhibitor of the endoplasmic reticulum Ca2+ pump) abolished 50 microM fluoxetine-induced Ca2+ release; whereas pretreatment with fluoxetine did not alter the thapsigargin-induced Ca2+ response. Addition of 3 mM Ca2+ increased [Ca 2+] i after pretreatment with 50 microM fluoxetine in Ca2+ -free medium, suggestive of fluoxetine-induced capacitative Ca2+ entry. Suppression of inositol 1,4,5-trisphosphate formation by 2 microM U73122 (a phospholipase C inhibitor) did not affect 50 microM fluoxetine induced Ca2+ release. Collectively, this study shows that fluoxetine increased [Ca 2+] i in bladder cancer cells in a concentration-dependent fashion, by releasing Ca2+ from thapsigargin-sensitive Ca2+ stores in an IP3-independent manner, and by inducing Ca2+ influx from extracellular medium. PMID- 11394945 TI - Prevention of ethanol-induced gastric lesions in rats by natural honey and glucose-fructose-sucrose-maltose mixture. PMID- 11394946 TI - Does a glucose-fructose-sucrose-maltose mixture (as present in honey) really possess gastric protection effects? PMID- 11394947 TI - The link between smoking and impotence: two decades of evidence. AB - BACKGROUND: Anti-tobacco advertisements now feature the risk of impotence as a reason to avoid or cease tobacco use. The scientific evidence, however, is mixed. To shed light on the controversy, we compiled 2 decades of published data on the link between smoking and impotence. METHODS: We searched MEDLINE from 1980 to the present for studies that reported smoking prevalence among impotent male subjects. For each study we recorded the age of subjects, their state of residence, the percentage that were current smokers, and the time period over which study data were collected. For comparison purposes, we estimated age-, state-, and year-specific smoking rates in the general male population using the Behavioral Risk Factors Surveillance System. We performed a meta-analysis using a random effects model. RESULTS: Among the 1008 journal articles examined, we identified 19 studies that reported the smoking habits of 3819 impotent men. Of these 19 studies, 16 indicated a smoking prevalence exceeding that of the general population. The 6 largest studies all revealed a higher prevalence of smoking among impotent men. Meta-analysis reveals that 40% of impotent men were current smokers compared with 28% of men in the general population. CONCLUSIONSBased on almost 2 decades of evidence, tobacco use is an important risk factor for impotence. Anti-tobacco advertisements featuring impotence as a reason to avoid or cease tobacco use are well grounded in scientific fact. PMID- 11394948 TI - Page for patients. Lyme disease. PMID- 11394949 TI - Factors associated with physical activity among Spanish youth through the National Health Survey. AB - BACKGROUND: Physical inactivity is emerging as one of the largest public health problems throughout Europe. This study updates descriptive data on physical activity and the factors associated with it among the 6- to 15-year-old Spanish population during leisure time through the National Health Survey. METHODS: This is a cross-sectional study from a representative sample of Spanish children's physical activity during their leisure time. Sample size was 1,358 children between the ages of 6 and 15 years. Univariate analyses were carried out first, and then significant variables were entered into a logistical regression nonconditional model. RESULTS: Less than 30% of children were active and the percentage of active boys was higher than that of active girls in all age categories. From the age of 11 the percentage of total active children was stabilized. Children from the two highest social statuses (SS) were significantly more active than children from the lowest SS group. Children from large towns were more active than children from small towns. Active children reported eating more meat or fish than the less active ones. CONCLUSIONS: The results suggest that characteristics of age, sex, town size, and social status should be considered when designing physical activity strategies for Spanish children. PMID- 11394950 TI - Effects of worksite physical exercise intervention on physical fitness, perceived health status, and work ability among home care workers: five-year follow-up. AB - OBJECTIVE: The effects and constancy of a worksite physical exercise intervention were examined in relation to the physical fitness, perceived health status, and work ability of female service workers during periods of 1 and 5 years. METHODS: The subjects comprised female home care workers divided into an intervention group (n = 50, mean age 41.8 (SD 10.4) years) and a control group (n = 37, mean age 43.3 (SD 8.8) years). The intervention group participated in 9 months of supervised exercise intervention twice a week during the workday. Functional capacity, perceived health, and work ability were assessed at the beginning of the study and after a 1- and a 5-year period of follow-up. RESULTS: In the 1-year follow-up measurements, body fat had decreased (4%) and dynamic muscle performance and maximal oxygen consumption in relation to body mass (30-38 and 7%, respectively) had increased in the intervention group. The differences in outcome variables between the intervention and the control groups were significant (from P = 0.014 to P < 0.001). These positive effects of worksite exercise were observed despite the age of the subjects, and the changes were consistent during a 5-year period. In the control group the decline of the work ability index (L smean) was about three times faster than in the intervention group during the 5-year period. CONCLUSIONS: Physical exercise executed in work units can be used to improve the physical capacity of female home care aides and prevent the early decline of their work ability. In jobs that are physically demanding, such as home care work, early prevention must start before the age related deterioration of health and physical capacity. PMID- 11394951 TI - Ecological and socioeconomic correlates of fruit, juice, and vegetable consumption among African-American boys. AB - BACKGROUND: Investigators have reported that the availability of foods in local grocery stores correlated with consumption when using large geopolitical units of analysis, e.g., zip codes. Associations across smaller geopolitical units, e.g., census tracts, have not been tested, nor has this work focused on restaurant availability, child consumption, or specific ethnic groups. METHODS: This study examined whether median family income and fruit, juice, and vegetable (FJV) availability in grocery stores, restaurants, and homes in 11 census tracts correlated with FJV consumption among 11- to 14-year-old African-American Boy Scouts. FJV consumption was measured in 90 scouts using two 24-h food recalls. Instruments were developed to measure the availability of FJV at area grocery stores, restaurants, and homes where troop members resided. RESULTS: Median household income (from 1990 census) was significantly correlated with restaurant fruit availability. Significant correlations were found between restaurant juice and vegetable availability and Boy Scout reported consumption of juice and vegetables. CONCLUSION: Census tract may be a useful unit when studying restaurant, but not grocery store, FJV availability. Within a census tract, restaurant FJV availability may be a significant target for community intervention and process evaluation. PMID- 11394952 TI - Reasons women do not attend screening for cervical cancer: a population-based study in Sweden. AB - BACKGROUND: With an improved compliance with screening a larger reduction of cervical cancer incidence would be within reach. We aimed at investigating why certain women do not attend Pap smear screening and at validating the reliability of self-reported screening. METHODS: In 1998 in the county of Uppsala, Sweden, information was collected through telephone interviews with 430 nonattendees and 514 attendees to Pap smear screening, who were all sampled from a population based database. The women's recall of attendance was validated against the database. The main outcome measures used were odds ratios (OR) and 95% confidence intervals (CI). RESULTS: Non-attendance was positively associated with nonuse of oral contraceptives (OR = 3.56, 95% CI 2.18-5.83), seeing different gynecologists (OR = 1.90, 95% CI 1.34-2.70), and seeing a physician very often (OR = 3.12, 95% CI 1.45-6.70) or not at all (OR = 1.78, 95% CI 1.09-2.90). Frequent condom use (OR = 1.88, 95% CI 1.02-3.47), living in rural/semirural areas (OR = 1.55, 95% CI 1.07-2.21), and not knowing the recommended screening interval (OR = 2.16, 95% CI 1.20-3.89) were all associated with nonattendance, whereas socioeconomic status was not, when tested in a multivariate model. Among the nonattendees, 57% underestimated the time lapse since last smear. CONCLUSIONS: Seeing a gynecologist on a regular basis and information guiding women to have a Pap smear on their own initiative are important factors for recurrent screening. Therefore, information should be given to all women about the purpose and benefits of Pap smear testing. Self-reports on screening should be treated with caution. PMID- 11394953 TI - Assessing the association of walking with health services use and costs among socioeconomically disadvantaged older adults. AB - BACKGROUND: The costs of physical inactivity are beginning to be recognized. Research to pinpoint these costs will provide needed information for researchers and policy-makers to develop cost-effective physical activity promotion programs. We present the association of walking with health services use and costs within a sample of 695 older, urban primary care patients. METHODS: A survey provided most data, but utilization and cost data were obtained from a medical records system. Multivariate models were developed to assess the association of walking with health services use and costs, adjusting for sociodemographic characteristics, chronic disease, health status, and previous utilization. RESULTS: Thirty-eight percent of respondents reported walking 0 minutes per week, 49% walked 1 to 119 minutes, and 13% walked 120 minutes or more. In the multivariate analyses, a report of walking 120 or more minutes was associated with a lower risk of emergency room visit and hospital stay in the subsequent year. No association was found between walking and primary care visits and total cost. CONCLUSION: These analyses suggest an association of walking 120 minutes or more with decreased emergency room visits (OR = 0.5, P = 0.046) and hospital stays (OR = 0.6, P = 0.034). This suggests that physical activity promotion among socioeconomically disadvantaged older adults has the potential to provide cost savings. This will not be known, however, until physical activity can be promoted and maintained among these adults. PMID- 11394954 TI - Risk and reluctance: understanding impediments to colorectal cancer screening. AB - PURPOSE: Screening to detect and prevent colorectal cancer (CRC) is well below optimal, contributing to needless CRC-related morbidity and mortality. Little detailed information exists explaining why screening technologies are underutilized and why screening adherence rates are low. Prior to the design of an intervention study, we assessed knowledge about CRC among adult women and men with access to health care. We also investigated patterns of perceived risk for CRC, barriers and facilitators to screening, and experience and intentions with regard to both fecal occult blood testing and flexible sigmoidoscopy. METHODS: We analyzed data from semistructured focus group interviews with a small, nonrepresentative sample (n = 39) of community-dwelling adult men and women ages 50 to 64 and 65 plus. RESULTS: CRC-related knowledge is low, and misperceptions are common. Provider practices reinforce low levels of perceived risk. Multiple barriers to screening exist, of which many are remediable. CONCLUSIONS: We are at an early stage in the diffusion of information about CRC. Screening utilization may be improved through development of appropriate public health awareness campaigns and by addressing service factors. Recommendations are provided. PMID- 11394955 TI - Project Towards No Drug Abuse: generalizability to a general high school sample. AB - OBJECTIVE: We examined the generalizability of a successful classroom-based prevention program developed for youth at alternative high schools (high risk) to youth at general high schools. METHOD: A replication of a previously tested prevention program in a general high school population was conducted with 1-year follow-up data. Classrooms within each of three schools were randomly assigned to two conditions, classroom education or standard care control. RESULTS: Statistically significant effects on alcohol and illicit drug use were achieved in this population through a 1-year period following the program, although effects were not achieved on cigarette smoking and marijuana use. CONCLUSIONS: These results suggest that this program (Project Towards No Drug Abuse) has applicability to a wide range of older teens. PMID- 11394956 TI - Dual Tobacco use among Native American adults in southeastern North Carolina. AB - OBJECTIVE: While patterns of smokeless tobacco (ST) use and cigarette smoking are well documented, the epidemiology of simultaneous use of both tobacco products is less well studied, particularly among Native American populations. This study examines correlates of dual tobacco use among Lumbee Indian adults in southeastern North Carolina. METHODS: A telephone survey among 400 adult Lumbee Indians in Pembroke, North Carolina, collected information on demographics, current tobacco use, amounts of tobacco used, and tobacco related attitudes. RESULTS: Total of 241 (60.3%) individuals did not currently use tobacco, 104 (26%) currently smoked, 74 (18.5%) currently used ST, and 19 (4.8%) used both products. Thus, 19 of 104 (18.3%) current smokers and 19 of 74 (25.7%) current ST users reported dual tobacco use. Compared to exclusive users of either tobacco product, dual tobacco users were intermediate in age and frequency of church attendance, had lower levels of education, and were the highest proportion of subjects reporting no friends and few close relatives. There was no difference by gender or marital status by tobacco use categories. While exclusive cigarette smokers reported smoking more cigarettes per day than dual tobacco users, overall, dual tobacco users had higher estimated daily nicotine exposure levels. Logistic regression analysis showed that younger age and infrequent church attendance predicted exclusive cigarette smoking, while older age and less education predicted exclusive ST use. Dual tobacco use was predicted only by less education. CONCLUSIONS: Simultaneous use of ST and cigarettes is comparatively more common among Lumbee Indian adults than the general population and has an epidemiology distinct from either exclusive cigarette smoking or ST use. These data are the first to explore social support as well as tobacco-related attitudes among dual tobacco users in a Native American population. Recognition of these patterns of dual tobacco use would be important in any future tobacco intervention among Lumbee Indian adults. PMID- 11394957 TI - Identifying and characterizing one-time and never users of mammography screening services among medicare eligible women. AB - BACKGROUND: Despite eligibility for a screening mammogram once every 2 years from 1991 to 1997, only a small percentage of Medicare women utilized this benefit. We examined mammography use among 388,707 North Carolina Medicare women from 1994 to 1997 to identify characteristics of one-time and never users of mammography. METHODS: Data were obtained from North Carolina Medicare mammography claims and enrollment files from 1994 to 1997. Women ages 65+ as of 01/01/1994, continuously enrolled in Medicare from 1994 to 1997, and alive as of 12/31/1997 were included in the sample (n = 388,707). Mammogram use was categorized as never, once, or at least twice during 1994/1995 and 1996/1997. Women with at least one mammography claim during 1994/1995 and at least one mammography claim during 1996/1997 were called repeat users, women with one mammography claim during the 4 years were labeled one-time users, and women with zero mammography claims during the 4 years were termed never users. Multivariate logistic regression analyses were conducted to determine associations between characteristics and mammography frequency. RESULTS: Biennial mammography claims data rates were 35.3% in 1994/1995 and 41.8% in 1996/1997. Compared with all other users, one-time users (n = 108,899) were more likely to be ages 65-74 (vs 75-84 and 85+), live in an urban versus rural county, and be eligible for Medicare only versus Medicare and Medicaid. Never users (n = 184,545) were more likely to be ages 85+, be non-Caucasian, live in a rural county, and be eligible for both Medicare and Medicaid versus Medicare. CONCLUSIONS: These results demonstrate different demographic characteristics for one-time and never mammography users. This approach of using multiple years of claims data to segment the targeted population provides the opportunity to tailor interventions to subgroups. PMID- 11394958 TI - The reaction of the immune system to pathogens but not food antigens and commensal bacteria. PMID- 11394959 TI - The role of epithelial cells in immune regulation in the gut. AB - A variety of mechanisms contribute to the ability of the gut to either react or remain tolerant to antigens present in the intestinal lumen. Intestinal epithelial cells can control the uptake, transmission and presentation of luminal antigens through an astonishingly diverse set of pathways. Antigens can cross the epithelial barrier via non-specific pinocytotic, specific receptor mediated, or intracellular/paracellular bypass pathways. The differential processing and presentation by a variety of restriction elements may result in the activation of functionally distinct target cell populations which have the capacity to regulate the predominant trend of immune unresponsiveness within the gut. PMID- 11394960 TI - Oral tolerance. AB - The ability of the mucosal immune system to distinguish between harmful and harmless antigens is essential for mounting protective immune responses and preventing the induction of mucosal pathology yet the basis for this remains unclear. As fed antigen can also exert systemic effects understanding oral tolerance and priming will also have important consequences for therapy and vaccination. Here we will not only review the increasing amount of information about the potential mechanisms of oral tolerance and priming but also attempt to shed some light on how differences in the uptake and handling i.e. 'the journey' of orally administered antigen may promote these mechanisms. PMID- 11394961 TI - Immuno-bacterial homeostasis in the gut: new insights into an old enigma. AB - The intestinal mucosa is the interface between the immune system and the massive antigenic load represented by the commensal enteric bacteria. These commensal bacteria drive the development of the mucosal immune system, and in turn most of the lymphocytes in the intestinal mucosa appear to be specific for enteric bacteria antigens. Proper regulation of the responses of these anti-bacterial lymphocytes are extremely important because T cell effectors reactive to enteric bacterial antigens have been shown to cause chronic intestinal inflammation in an adoptive transfer system. The cells and molecules important in regulating mucosal immune response are now being identified. Insights into the mechanisms of mucosal regulation have come from a number of genetically manipulated mouse strains which develop inflammatory bowel disease in response to the enteric bacterial flora. CD4(+)T cells with regulatory function in the mucosa are being identified; other cell types such as CD8(+)T cells. NK cells, and B cells may also have a role in mucosal immune regulation. A model for T cell-immune homeostasis in the intestinal mucosa is presented. PMID- 11394962 TI - Regulation of life and death in lamina propria T cells. AB - T cells are essential to initiation, amplification, and regulation of an immune response. This response is terminated when T cells undergo apoptosis, a physiological process of cell death triggered by various mechanisms and regulated by signaling pathways leading to enzymatic degradation of chromatin. An effective immune response depends on the proper balance between proliferation and death of activated T cells. This is particularly important in the intestine, where mucosal T cells are subjected to the high antigenic pressure of lumenal antigens and apoptosis is required to induce tolerance and maintain a state of 'physiological' inflammation. Insufficient apoptosis may result in excessive T cell retention and chronic intestinal inflammation, as seen in conditions associated with defective apoptosis of lamina propria T cells. PMID- 11394963 TI - Understanding mucosal responsiveness: lessons from enteric bacterial pathogens. AB - Mucosal immune responses must discriminate between commensal flora within the lumen and potential pathogens. These responses are highly adapted to induce protection without excessive inflammation. The balances that regulate mucosal immune and inflammatory responses have to be understood if effective mucosal immunity is to be induced through local immunization. This review will summarize some of the lessons learnt from studies of antigens derived from enteric bacterial pathogens and discuss how the gastrointestinal epithelia can 'fight back' when it encounters pathogens. PMID- 11394965 TI - Site-specific mutations of FtsZ--effects on GTPase and in vitro assembly. AB - BACKGROUND: FtsZ, the major cytoskeletal protein in bacterial cytokinesis, assembles in vitro into protofilaments, which can further associate into sheets, bundles or tubes. We have constructed 16 site-directed mutants of E. coli ftsZ, and tested them for GTP hydrolysis and assembly in vitro, and for their ability to complement the temperature sensitive ftsZ84 mutation in E. coli. RESULTS: The mutants were grouped into three classes. Benign mutants, which mapped mostly to the front and back surface of the protofilament, were able to complement ftsZ84 in vivo and showed normal assembly in vitro. GTP contact mutations had less than 10% of wild type GTPase activity. They could all assemble in vitro, and several of these mutants could complement ftsZ84. A third, and newly discovered, class of mutations mapped to the sides of the protofilaments. These lateral mutants had mostly normal GTPase and assembly in vitro, but none of them complemented ftsZ84. The non-complementing mutants showed greatly reduced expression from the pBS58 vector, suggesting possible dominant negative effects. CONCLUSIONS: Several mutants with greatly reduced GTPase could still complement ftsZ84, suggesting that the high level of GTPase observed in vitro is not essential for in vivo function. All of the lateral mutants failed to complement ftsZ84, which suggests that these surfaces of the protofilaments are important for function in cell division. These lateral surfaces may mediate association of FtsZ protofilaments into pairs or small sheets, although their structure is apparently different from the sheets assembled in DEAE dextran or calcium. PMID- 11394966 TI - Intestinal parasitic infections in Thai HIV-infected patients with different immunity status. AB - BACKGROUND: One of the major health problems among HIV seropositive patients is superimposed infection due to the defect of immunity. Furthermore, intestinal parasite infection, which is also one of the basic health problems in tropical region, is common in these patients. In this study, a cross sectional study to document the prevalence of intestinal parasitic infection in Thai HIV-infected patients with different immune status was performed. METHODS: A study of stool samples from 60 Thai HIV-infected patients with different immune status was performed at King Chulalongkorn Memorial Hospital, Thailand. Each patient was examined for CD4 count and screened for diarrheal symptoms. RESULTS: The prevalence of intestinal parasitic infection among the HIV-infected patients in this study was 50 %. Non- opportunistic intestinal parasite infections such as hookworms, Opisthorchis viverrini and Ascaris lumbricoides were commonly found in HIV-infected people regardless of immune status with or without diarrheal symptoms. Opportunistic intestinal parasites such as Cryptosporidium, Isospora belli, Microsporidia and Strongyloides stercoralis infection were significantly more frequent in the low immunity group with diarrhea. CONCLUSION: Therefore, opportunistic intestinal parasite infection should be suspected in any HIV infected patient with advanced disease presenting with diarrhea. The importance of tropical epidemic non-opportunistic intestinal parasite infections among HIV infected patients should not be neglected. PMID- 11394964 TI - Rhythmic expression of Nocturnin mRNA in multiple tissues of the mouse. AB - BACKGROUND: Nocturnin was originally identified by differential display as a circadian clock regulated gene with high expression at night in photoreceptors of the African clawed frog, Xenopus laevis. Although encoding a novel protein, the nocturnin cDNA had strong sequence similarity with a C-terminal domain of the yeast transcription factor CCR4, and with mouse and human ESTs. Since its original identification others have cloned mouse and human homologues of nocturnin/CCR4, and we have cloned a full-length cDNA from mouse retina, along with partial cDNAs from human, cow and chicken. The goal of this study was to determine the temporal pattern of nocturnin mRNA expression in multiple tissues of the mouse. RESULTS: cDNA sequence analysis revealed a high degree of conservation among vertebrate nocturnin/CCR4 homologues along with a possible homologue in Drosophila. Northern analysis of mRNA in C3H/He and C57/Bl6 mice revealed that the mNoc gene is expressed in a broad range of tissues, with greatest abundance in liver, kidney and testis. mNoc is also expressed in multiple brain regions including suprachiasmatic nucleus and pineal gland. Furthermore, mNoc exhibits circadian rhythmicity of mRNA abundance with peak levels at the time of light offset in the retina, spleen, heart, kidney and liver. CONCLUSION: The widespread expression and rhythmicity of mNoc mRNA parallels the widespread expression of other circadian clock genes in mammalian tissues, and suggests that nocturnin plays an important role in clock function or as a circadian clock effector. PMID- 11394967 TI - Retrospective confidence judgements made by adults with traumatic brain injury: relative and absolute accuracy. AB - Eighteen adult survivors of moderate or severe traumatic brain injury (TBI) and 18 controls studied and were tested on two lists of noun-pairs, and made item-by item retrospective confidence judgements (RCJs) of their answers using a Likert scale. For both groups, correlations between RCJ ratings and recall were extremely high, indicating that survivors of TBI were as accurate as non-injured controls when comparing their confidence in one answer to another. However, as groups, survivors of TBI were over-confident and control participants were under confident, but only when uncertain as to the accuracy of their answer. Recall errors of 'interference' were analysed post-hoc. Survivors of TBI displayed higher confidence in interference errors than in non-interference errors, while control participants did not. These results are discussed within the context of self-monitoring by clinical and non-clinical populations. The importance of both relative and absolute measures of metamemory accuracy is highlighted. PMID- 11394968 TI - Prediction of neuropsychiatric outcome following mild trauma brain injury: an examination of the Glasgow Coma Scale. AB - The relationship between the Glasgow Coma Scale (GCS) and neuropsychiatric outcome was examined in 57 consecutive subjects with mild traumatic brain injury (TBI) attending a follow-up clinic. Subjects were grouped according to initial GCS score (15 versus 13-14) and contrasted at an average of 5-6 months post injury. As expected, those with GCS 13-14 had longer PTA (p = 0.001) and a higher rate of abnormal brain CT scans (p = 0.005). However, no significant differences emerged for indices of neuropsychiatric status, including measures of neurobehavioural symptoms/signs, overall psychological distress, psychiatric 'caseness', functional and psychosocial outcome, frequency of common somatic complaints, and rate of return to work. Subsidiary analyses based upon the presence/absence of CT abnormalities and the duration of PTA (<1 hour versus 1-24 hours) also failed to predict outcome, although a trend associating longer PTA with lower functional outcome was observed. Thus, despite early neurosurgical differences, the results suggest that initial GCS scores do not clearly translate into neuropsychiatric sequelae at follow-up within the rubric of GCS 13-15. PMID- 11394969 TI - The utility of the Substance Abuse Subtle Screening Inventory-3 for use with individuals with brain injury. AB - The utility of the Substance Abuse Subtle Screening Inventory (SASSI-3) for the use with individuals with traumatic brain injuries (TBI) was investigated. The SASSI-3 was administered to 78 subjects prior to discharge from inpatient rehabilitation. The SASSI-3 diagnosis of chemical dependency was compared with the diagnosis of staff psychologists. Lower accuracy, sensitivity, and specificity were found in SASSI-3 diagnosis for the subjects with brain injury, as compared to a normative sample of persons with disabilities participating in a vocational rehabilitation programme. When a diagnosis of 'dependence or abuse' was used, accuracy of the SASSI-3 increased. Comparison of diagnosis based on SASSI-3 versus Blood Alcohol Level (BAL) at the time of injury showed comparable accuracy; however, BAL was found to have higher specificity. For dependence or abuse, BAL continued to be more specific than the SASSI-3; however, the SASSI-3 was more sensitive. Based on these findings, a clinical approach to screening using both BAL and the SASSI-3 is discussed. PMID- 11394970 TI - Circadian rhythm of temperature in head injury. AB - It has been shown in a previous study that head injured patients appear to have a circadian rhythm of their body functions. This needed to be confirmed using additional data, better collection methods and analysis. Additional goals were to develop a method of detrending of physiological time series in order to improve rhythm detection when it may be hidden behind a low frequency trend and the creation of a computer system for data acquisition and analysis. The temperature data of 10 head injured patients was studied using the Iterative Cosinor method. In one case, prior to the Cosinor method, detrending of the data was used using a specially designed polynomial fitting technique. The Iterative Cosinor method showed circadian rhythms in nine out of 10 patients. After detrending, a rhythm was found in the data of the 10th patient as well. The periods of the rhythm were around, but were not equal to, 24 hours. The results show that comatose head injured patients have a circadian rhythm of their core temperature. The detection of a circadian rhythm may, in some cases, be improved by using a detrending technique. The deviation of the rhythm period from 24 hours suggests that the rhythms found in these patients are free-running, meaning that head injured patients are not synchronized with their surroundings. PMID- 11394971 TI - Mathematical models of cognitive recovery. AB - Longitudinal psychological test results are used as dependent variables to explore the complex relationship between length of coma, time of testing on the recovery curve, and corresponding cognitive status after traumatic brain injury (TBI). A database containing 319 TBI patients with a broad spectrum of coma duration was used. Statistical analysis of mixed effects modelling was applied to longitudinal WAIS-R (Wechsler Adult Intelligence Scale-Revised) scores to construct two mathematical models (verbal IQ and performance IQ). The models predict the course of recovery (initial cognitive level post-coma, eventual recovery level, and level of cognitive functioning at any point on the recovery curve) when the duration of coma is known. Performance IQ was found to recover at a rate that is almost four times slower than verbal IQ. The results have important clinical rehabilitation implications. This statistical modelling technique also enables the medical researcher to investigate disease progression or recovery using structured assessments, which would normally be part of the routine medical monitoring. PMID- 11394972 TI - Tracheotomy in severe TBI patients: sequelae and relation to vocational outcome. AB - The aim of the present study was to find the influence of performing tracheotomy on outcome of severe TBI patients. TBI patients, many of them intubated during the very acute phase post-injury, who remain unconscious for more than a few days, undergo tracheotomy to provide a reliable long term artificial airway. Tracheotomy, although being a simple elective surgical procedure, may have a negative influence on the rehabilitation process. Tracheotomy was performed in 25% (n=69) of 277 consecutive severe TBI patients admitted to the rehabilitation department during 3 years. In seven patients (10%), tracheotomy was performed immediately after injury due to cranio-facial trauma, in 18 patients (29%) due to respiratory disturbances, in 42 patients (55%) after prolonged intubation, and in two patients (6%) due to combined problems. Forty-five per cent of patients with tracheotomy suffered from respiratory disturbances and 8.6% from complications of tracheotomy. Longer periods of unconsciousness and mechanical ventilation were significantly associated with respiratory complications (p<0.0001 and p<0.001, respectively). However, presence of tracheotomies per se, did not affect vocational rehabilitation. PMID- 11394973 TI - Head injuries in men's and women's lacrosse: a 10 year analysis of the NEISS database. National Electronic Injury Surveillance System. AB - INTRODUCTION: Although protective headgear is required in men's lacrosse, women's lacrosse is viewed as non-contact, and use of helmets and faceguards is prohibited. Yet, women remain at risk for injury to the head and face region from contact with the ball and stick. This study was designed to examine differences in lacrosse-related injuries between genders and amongst various age groups. METHODS: Data on lacrosse-related injuries maintained through the Consumer Product Safety Commission's National Electronic Injury Surveillance System, In depth Investigation File, and Injuries/Potential Injuries File were analysed over a 10-year period (January 1990-April 2000). RESULTS: A total of 1727 cases of lacrosse-related trauma, mean age 16.9 years, range 4-59 years, were recorded. Males accounted for 80.5% of cases. The head and face region was the most common area injured (20.4%). Injuries to the head and face were significantly more prevalent among females (30.1% of all injuries) than males (18.0% of all injuries), p < 0.001, and often resulted from contact with the ball (33.6% of incidents). Children aged 4-11 years experienced the highest percentage of injuries to the head and face. Closed head injuries represented 5.6% of all lacrosse-related injuries and were slightly more prevalent among females. CONCLUSIONS: Women and children lacrosse players are at risk of serious injury to the head and face region. The use of protective head/face gear should be encouraged. PMID- 11394974 TI - Effects of TENS and methylphenidate in tuberculous meningo-encephalitis. AB - PRIMARY OBJECTIVE: Beneficial effects of transcutaneous electrical nerve stimulation (TENS) on cognition and behaviour were observed in a child with probable Herpes Simplex Encephalitis. Based on these positive findings, it was examined in the present case study whether a child who had been diagnosed to suffer from tuberculous meningitis would benefit from TENS. Furthermore, as aggression and overactive behaviour were also prominent clinical symptoms, the effects of methylphenidate were investigated. METHODS AND PROCEDURES: Neuropsychological tests were used to assess attention/concentration and visuospatial and visuoconstructive memory. Behaviour, including the level of activity during 24 hours, was assessed by one observation scale and actigraphy. EXPERIMENTAL INTERVENTIONS: TENS and methylphenidate. MAIN OUTCOMES AND RESULTS: TENS particularly improved overall affective behaviour. Methylphenidate appeared to have the opposite effect on cognition and hardly any effect on patient's behaviour. CONCLUSIONS: TENS might improve the patient's behavioural functioning. Pros and cons for treatment effects are discussed. PMID- 11394975 TI - Vaccine candidates in STD. AB - Sexually transmitted diseases (STDs) are caused by organisms that infect the mucosal surfaces of the genitourinary tract. In spite of its public health importance, particular scientific problems have delayed the development of an STD vaccine, such as incomplete attenuation (human herpes simplex virus type 2), accentuated immunopathology (Chlamydia trachomatis), poor immunogenicity (Treponema pallidum), and broad antigenic heterogeneity (Neisseria gonorrhoeae). Nevertheless, efforts continue with the use of protein antigens: for example, the haemolysin toxoid of Haemophilus ducreyi; the major outer membrane protein(s) of N. gonorrhoeae and C. trachomatis; the glycoprotein D of human herpes simplex virus type 2; and the proteins E6 and E7 of the human papillomavirus. It could be predicted that eventual STD vaccines (administered either for prophylaxis or for therapy) will use approaches that will include (1) live-attenuated viruses, (2) subunit proteins or inactivated whole organisms given with mucosal adjuvants or with cellular immune response adjuvants, or (3) DNA plasmids expressing the vaccine antigen. PMID- 11394976 TI - Donovanosis: an update. AB - Donovanosis has been ignored for many years until recently. The condition still has a limited geographical distribution. A significant epidemic of donovanosis has been identified in KwaZulu/Natal, South Africa where it may be a risk factor for acquiring HIV in men. After a gap of more than 30 years, the organism was cultured by researchers in Durban, South Africa and Darwin, Australia. Polymerase chain reaction (PCR) techniques for donovanosis were developed soon after, most recently using a colorimetric detection system. Similarities between the causative organism, Calymmatobacterium granulomatis and Klebsiella spp. were confirmed. A proposal that the organism be reclassified under the genus Klebsiella has been put forward. Azithromycin has been confirmed as the drug of choice but is yet to be accepted universally because of cost issues. Treatment in patients with significant HIV induced immune deficiency may need to be prolonged. A donovanosis eradication programme is underway amongst the aboriginal community in Australia. Elsewhere, management through current syndromic guidelines for genital ulcers are yet to be validated in areas where donovanosis is endemic. PCR testing should enable further recognition of donovanosis and lead to more concerted efforts in disease control and possible eradication. PMID- 11394977 TI - Temperature stability of vaginal specimens for Chlamydia trachomatis detection by Amplicor polymerase chain reaction assay. AB - Vaginal introital specimens were collected from 17 women - 10 positive and 7 negative for Chlamydia trachomatis, and kept in Amplicor collection medium at ambient temperature. Aliquots were removed at intervals for up to 34 days and frozen at -80 degrees C. Samples were thawed and assayed for C. trachomatis by polymerase chain reaction (PCR). The results of all specimens remained unchanged over this time interval. PMID- 11394978 TI - Screening for Chlamydia trachomatis infection using the BDProbeTec ET Chlamydia trachomatis amplified DNA assay on urine in a GUM clinic setting: a simple, fast and cost-effective alternative. AB - This study compared the BDProbeTec ET Chlamydia trachomatis amplified DNA assay on urine specimens with culture of genital swabs for the detection of C. trachomatis in patients attending the Department of Genitourinary Medicine (GUM), Cardiff Royal Infirmary. Almost twice as many patients tested positive by BDProbeTec ET than by culture. A similar difference was found for both males and females. The case notes of those patients positive by BDProbeTec ET alone were analysed and a significantly greater number were found to have risk indicators for C. trachomatis infection when compared with age and sex comparable controls, providing clinical validation of our findings. The BDProbeTec ET assay was easy to use, more importantly, the test format features an internal control integral with every sample. The cost per true positive was calculated as comparable with culture. We conclude that the BDProbeTec ET assay is a superior alternative to culture for identifying patients infected with C. trachomatis in the GUM clinic setting. PMID- 11394979 TI - Characteristics of homosexually-active men with gonorrhoea during an epidemic in Sydney, Australia. AB - Our objective was to investigate an epidemic of gonorrhoea among homosexually active men in Sydney. Demographic and behavioural data on all homosexually-active men diagnosed with gonorrhoea (any site) at the Sydney Sexual Health Centre (SSHC) from 1992 through 1998 were reviewed. The men diagnosed with anal gonorrhoea were then compared with all homosexually-active men who tested negative for anal gonorrhoea or who were not tested for anal gonorrhoea at the SSHC between 1996 and 1998. Data on HIV status and country of birth of men diagnosed with anal gonorrhoea during 1998 at the Taylor Square Private Clinic were also reviewed. Over the period 1992 to 1998, homosexually-active men diagnosed with gonorrhoea at SSHC tended to become older at the time of diagnosis (median age 26.5 years in 1992 up to 31.0 years in 1998), indicating a cohort effect in the clinic population due to service reductions. When compared with men who tested negative for anal gonorrhoea at SSHC between 1996 and 1998, those with anal gonorrhoea were more likely to have anogenital symptoms (adjusted odds ratio [OR] 2.3), to have had a past history of gonorrhoea (OR 3.1), to present as a contact of gonorrhoea (OR 8.6), to have used condoms less consistently (OR 2.3), to be HIV positive or of unknown HIV status (OR 3.2), and to have been born in an English-speaking country other than Australia (OR 2.9). The last feature was not observed at the private clinic. In conclusion, the gonorrhoea epidemic was linked to public health service reductions, though it seems unlikely to be the only factor. Homosexually-active men with anal gonorrhoea had well established behavioural risk factors while men with concurrent HIV infection were over represented. Given the role of gonorrhoea in promoting the spread of HIV infection, a National Sexual Health Strategy--closely linked to the National HIV/AIDS Strategy--is due. PMID- 11394980 TI - Management of sexually transmitted diseases in urban pharmacies in The Gambia. AB - This study aimed to describe the quality and costs of sexually transmitted disease (STD) case management in urban pharmacies in The Gambia, and explore pharmacy workers' (PWs) willingness to improve the STD care they provide. PWs from 24 registered pharmacies were interviewed in order to collect information on their knowledge and practices regarding management of STDs. The same pharmacies were visited by a male 'simulated client' (SC) to ascertain how urethral discharge syndrome (UDS) cases were managed in practice. Fifteen (63%) pharmacies were equipped for treatment of UDS, pelvic inflammatory disease (PID) and genital ulcer syndrome (GUS), according to national guidelines. Appropriate syndromic management for UDS was mentioned by 11% of PWs but actually given to 4.4% of the SC visits. None of the PID or GUS cases would be treated correctly. Forty-two per cent of PWs advised on partner notification, 38% on safe sex and 29% on treatment compliance in the SC visits. The reported costs for treatment of UDS, PID and GUS ranged from $2.5-$15.0. The cost of treatment actually purchased by the SC averaged $3.5 (range $1.5-$9.6) for UDS. Excluding the pharmacy sector from interventions will limit the impact of STD control measures. Regular training in syndromic management and rational drug use, with a concise manual for reference are recommended. Strategies to lower the cost of drugs should be explored. PMID- 11394981 TI - Changing patterns of presentations of patients with HIV-related disease at a tertiary referral centre and its implications for physician training. AB - Highly active antiretroviral therapy (HAART) has been shown to be highly effective in controlling HIV-related disease progression. Our objective was to determine whether HAART had altered the spectrum of HIV-related disease presentations at a tertiary medical referral centre and if a change in the clinical presentations of HIV-infected individuals to the hospital had impacted on physicians' training. A retrospective study which examined all admissions of HIV-infected patients identified between 1 October 1996 to 30 September 1998 using a hospital-designed computer database was undertaken at the Beth Israel Deaconess Medical Center (BIDMC) tertiary medical referral centre. All medical residents were surveyed in order to assess their knowledge of HIV-associated admissions and their confidence treating HIV-infected patients. There were significant changes in the admitting diagnosis for HIV-related illness between 1996 and 1998. Admissions for opportunistic infections (OIs) declined whereas admissions with bacterial infections increased significantly. Use of HAART remained stable between the 2 years of the study. Physicians' overestimated the use of HAART and only 8% of residents felt very comfortable taking care of an HIV infected patient. In conclusion, the spectrum of presentations with HIV-related disease to a tertiary referral centre continues to change in the HAART era and impacts on physicians' experience of the management of HIV disease. PMID- 11394982 TI - Stabilization of HIV infection rates in urban Burkina Faso, 1995-1999. AB - The objectives of this study were to monitor the trends of the HIV epidemic between 1995 and 1999 among pregnant women in Bobo-Dioulasso, the second largest town of Burkina Faso, and to discuss the possible effect of preventive interventions (condom availability) on sexual transmission of HIV in this context. Age-specific trends in HIV prevalence obtained from sentinel surveillance programme were analysed. Among antenatal clinic attendees, HIV prevalence was 7.5% (n=401) in 1995, 10% (n=200) in 1996, 7.6% (n=448) in 1997, 8.4% (n=642) in 1998 and 5.3% (n=716) in 1999 without demonstrated temporal trend (P=0.12). The average number of condoms available per person (aged 15-49 years) per year increased from 0.6 in 1992 to 5.7 in 1995 and 6.0 in 1999. Anonymous surveys are less subject to selection bias and suggest a stabilization of the HIV prevalence around 7.3% in Bobo-Dioulasso. Distribution of condoms could explain at least, partly, this stabilization of the HIV epidemic. PMID- 11394983 TI - Dyslipidaemia in HIV-infected patients: association with adherence to potent antiretroviral therapy. AB - Metabolic complications are being increasingly recognized among HIV-infected patients treated with potent combination antiretroviral therapies. We sought to assess the association of dyslipidaemia with adherence to protease inhibitor (PI) therapy and with the markers of clinical response to antiretroviral therapy (CD4 count, HIV RNA viral level) through a prospective, cross-sectional cohort study. Fifty-six HIV-infected patients who were already on, or who were started on PI containing antiretroviral therapy were monitored for the development of dyslipidaemias. Therapy with PI-containing antiretroviral therapy was significantly associated with elevated serum triglyceride level (>250 mg/dl) (52% vs 8%, P=0.001). Patients with an adherence rate of at least 80% to a PI containing regimen were significantly more likely to have elevated low density lipoprotein (LDL) cholesterol level as compared to patients with an adherence rate of <80% (79% vs 26%, P=0.03). Patients with an adherence rate of at least 80% to a PI-containing regimen were also significantly more likely to have severe hypertriglyceridaemia (>800 mg/dl) as compared to patients with an adherence rate of <80% (21% vs 4%, P=0.04). Viral load at the last study visit did not correlate with total cholesterol (r=-0.39, P=0.30), LDL cholesterol (r=0.57, P=0.30), or triglyceride level (r=0.55, P=0.20). However, there was a significant correlation between the last viral load and high density lipoprotein (HDL) cholesterol (r=0.79, P=0.035), i.e. lower viral load was associated with higher HDL cholesterol level. In conclusion, dyslipidaemia in patients with HIV infection was significantly associated with adherence to PI-containing antiretroviral therapy. Patients who are adherent to PI-containing regimens at least 80% of the time warrant close monitoring for the development of dyslipidaemia. PMID- 11394984 TI - Condom use during a community intervention trial in Kenya. AB - We conducted a cluster-randomized community intervention trial at Kenyan agricultural sites to measure the impact of female condom introduction on sexually transmitted infection (STI) prevalence. We present male and female condom use data here. Six Intervention sites received a community risk-reduction campaign and distribution of female condoms and male condoms, while 6 Control sites received the same campaign with male condoms only. Male and female condom distribution increased throughout follow-up. Self-reported male condom use increased substantially during follow-up to over 60% of the participants. The proportion of consistent male condom users at Control sites was higher than at Intervention sites, 23% vs 14% at 6 months and 24% vs 22% at 12 months. At Intervention sites, 11% and 7% of women used the female condoms all the time at 6 and 12 months, respectively, while the percentage of female condom non-users grew. Male and female condom use was hindered by male partner objections; suspicion of the study and the devices among residents; and bias against condoms by clinic service providers. A large proportion of coital acts remained unprotected during the trial. Our female condom intervention did not reduce STI prevalence, compared with male condom promotion only. PMID- 11394985 TI - 'Condom club': an interface between teenage sex and genitourinary medicine. AB - In response to demand from young teenagers, a nurse-led 'condom club' was established in 1996. This paper reports the use of the service from 1996 to 1999 by retrospective case note review. Two hundred and eighty-six individual males and 327 individual females made 655 and 824 visits, respectively. Condoms were accessed by 495 individuals, but girls also requested emergency contraception and pregnancy testing. Young teenagers were reluctant to have screening for sexually transmitted infections (STIs), but in those that did, 50% were found to have an STI. It is concluded that young teenagers will access a nurse-led sexual health advice service. The use of nursing staff allows greater flexibility of access. Geographical accessibility was also shown to be important. PMID- 11394986 TI - Penile fracture. AB - A wide range of patients present to genitourinary medicine (GUM) clinics with diverse conditions, many of which may not be infective. One such condition, albeit rare, is penile fracture. This is a traumatic rupture of the corpus cavernosum usually associated with blunt trauma to the erect penis. Prognosis and management issues are discussed. PMID- 11394987 TI - Gynaecomastia in a male patient during stavudine and didanosine treatment for HIV infection. AB - We report a case of gynaecomastia developed in a HIV-seropositive man, associated with a severe lipodystrophy. We hypothesize the responsibility of stavudine and didanosine in the development of these 2 complications. If many reports suggest that the protease inhibitors may promote gynaecomastia, long-term nucleoside analogue therapy may also cause this side effect. PMID- 11394989 TI - Chlamydia trachomatis detection: is it doctor dependent? PMID- 11394990 TI - Lim homeobox genes and the CNS: a close relationship. PMID- 11394991 TI - Is the retina going digital? PMID- 11394992 TI - Unlocking the DOR code. PMID- 11394993 TI - Smelling well with a code in the nodes. PMID- 11394994 TI - Challenging the neurocentric view of neuromuscular synapse formation. PMID- 11394995 TI - The cortical distribution of sensory memories. PMID- 11394996 TI - The prefrontal cortex--an update: time is of the essence. PMID- 11394997 TI - Organizing principles of the axoglial apparatus. AB - On axonal surfaces that flank the node of Ranvier and in overlying glial paranodal loops, proteins are arranged within circumscribed microdomains that defy explanation by conventional biosynthetic mechanisms. We postulate that the constraint of proteins to these loci is accomplished in part by discriminative membrane-embedded molecular sieves and diffusion barriers, which serve to organize and redistribute proteins after delivery by vesicular transport to neural cell plasma membranes. One sieve likely comprises a moveable, macromolecular scaffold of axonal and glial cell-derived transmembrane adhesion molecules and their associated cytoplasmic binding partners, located at the ends of each elongating myelin internode; this sieve contributes to restricting the sodium channel complexes to the node. We also anticipate the existence of a passive paranodal diffusion barrier at the myelin/noncompact membrane border, which prohibits protein diffusion out of contiguous paranodal membranes. PMID- 11394998 TI - Genetic ablation of orexin neurons in mice results in narcolepsy, hypophagia, and obesity. AB - Orexins (hypocretins) are a pair of neuropeptides implicated in energy homeostasis and arousal. Recent reports suggest that loss of orexin-containing neurons occurs in human patients with narcolepsy. We generated transgenic mice in which orexin-containing neurons are ablated by orexinergic-specific expression of a truncated Machado-Joseph disease gene product (ataxin-3) with an expanded polyglutamine stretch. These mice showed a phenotype strikingly similar to human narcolepsy, including behavioral arrests, premature entry into rapid eye movement (REM) sleep, poorly consolidated sleep patterns, and a late-onset obesity, despite eating less than nontransgenic littermates. These results provide evidence that orexin-containing neurons play important roles in regulating vigilance states and energy homeostasis. Orexin/ataxin-3 mice provide a valuable model for studying the pathophysiology and treatment of narcolepsy. PMID- 11394999 TI - EGR2 mutations in inherited neuropathies dominant-negatively inhibit myelin gene expression. AB - The identification of EGR2 mutations in patients with neuropathies and the phenotype Egr2/Krox20(-/-) have demonstrated that the Egr2 transcription factor is critical for peripheral nerve myelination. However, the mechanism by which these mutations cause disease remains unclear, as most patients present with disease in the heterozygous state, whereas Egr2(+/-) mice are phenotypically normal. To understand the effect of aberrant Egr2 activity on Schwann cell gene expression, we performed microarray expression profiling to identify genes regulated by Egr2 in Schwann cells. These include genes encoding myelin proteins and enzymes required for synthesis of normal myelin lipids. Using these newly identified targets, we have shown that neuropathy-associated EGR2 mutants dominant-negatively inhibit wild-type Egr2-mediated expression of essential myelin genes to levels sufficiently low to result in the abnormal myelination observed in these patients. PMID- 11395000 TI - Axon-glia interactions and the domain organization of myelinated axons requires neurexin IV/Caspr/Paranodin. AB - Myelinated fibers are organized into distinct domains that are necessary for saltatory conduction. These domains include the nodes of Ranvier and the flanking paranodal regions where glial cells closely appose and form specialized septate like junctions with axons. These junctions contain a Drosophila Neurexin IV related protein, Caspr/Paranodin (NCP1). Mice that lack NCP1 exhibit tremor, ataxia, and significant motor paresis. In the absence of NCP1, normal paranodal junctions fail to form, and the organization of the paranodal loops is disrupted. Contactin is undetectable in the paranodes, and K(+) channels are displaced from the juxtaparanodal into the paranodal domains. Loss of NCP1 also results in a severe decrease in peripheral nerve conduction velocity. These results show a critical role for NCP1 in the delineation of specific axonal domains and the axon glia interactions required for normal saltatory conduction. PMID- 11395001 TI - Contactin orchestrates assembly of the septate-like junctions at the paranode in myelinated peripheral nerve. AB - Rapid nerve impulse conduction depends on specialized membrane domains in myelinated nerve, the node of Ranvier, the paranode, and the myelinated internodal region. We report that GPI-linked contactin enables the formation of the paranodal septate-like axo-glial junctions in myelinated peripheral nerve. Contactin clusters at the paranodal axolemma during Schwann cell myelination. Ablation of contactin in mutant mice disrupts junctional attachment at the paranode and reduces nerve conduction velocity 3-fold. The mutation impedes intracellular transport and surface expression of Caspr and leaves NF155 on apposing paranodal myelin disengaged. The contactin mutation does not affect sodium channel clustering at the nodes of Ranvier but alters the location of the Shaker-type Kv1.1 and Kv1.2 potassium channels. Thus, contactin is a crucial part in the machinery that controls junctional attachment at the paranode and ultimately the physiology of myelinated nerve. PMID- 11395002 TI - Patterning of muscle acetylcholine receptor gene expression in the absence of motor innervation. AB - The patterning of skeletal muscle is thought to depend upon signals provided by motor neurons. We show that AChR gene expression and AChR clusters are concentrated in the central region of embryonic skeletal muscle in the absence of innervation. Neurally derived Agrin is dispensable for this early phase of AChR expression, but MuSK, a receptor tyrosine kinase activated by Agrin, is required to establish this AChR prepattern. The zone of AChR expression in muscle lacking motor axons is wider than normal, indicating that neural signals refine this muscle-autonomous prepattern. Neuronal Neuregulin-1, however, is not involved in this refinement process, nor indeed in synapse-specific AChR gene expression. Our results demonstrate that AChR expression is patterned in the absence of innervation, raising the possibility that similarly prepatterned muscle-derived cues restrict axon growth and initiate synapse formation. PMID- 11395003 TI - Proprioceptor pathway development is dependent on Math1. AB - The proprioceptive system provides continuous positional information on the limbs and body to the thalamus, cortex, pontine nucleus, and cerebellum. We showed previously that the basic helix-loop-helix transcription factor Math1 is essential for the development of certain components of the proprioceptive pathway, including inner-ear hair cells, cerebellar granule neurons, and the pontine nuclei. Here, we demonstrate that Math1 null embryos lack the D1 interneurons and that these interneurons give rise to a subset of proprioceptor interneurons and the spinocerebellar and cuneocerebellar tracts. We also identify three downstream genes of Math1 (Lh2A, Lh2B, and Barhl1) and establish that Math1 governs the development of multiple components of the proprioceptive pathway. PMID- 11395004 TI - Functional repression of Islet-2 by disruption of complex with Ldb impairs peripheral axonal outgrowth in embryonic zebrafish. AB - Islet-2 is a LIM/homeodomain-type transcription factor of the Islet-1 family expressed in embryonic zebrafish. Two Islet-2 molecules bind to the LIM domain binding protein (Ldb) dimers. Overexpression of the LIM domains of Islet-2 or the LIM-interacting domain of Ldb proteins prevented binding of Islet-2 to Ldb proteins in vitro and caused similar in vivo defects in positioning, peripheral axonal outgrowth, and neurotransmitter expression by the Islet-2-positive primary sensory and motor neurons as the defects induced by injection of Islet-2-specific antisense morpholino oligonucleotide. These and other experiments, i.e., mosaic analysis, coexpression of full-length Islet-2, and overexpression of the chimeric LIM domains derived from two different Islet-1 family members, demonstrated that Islet-2 regulates neuronal differentiation by forming a complex with Ldb dimers and possibly with some other Islet-2-specific cofactors. PMID- 11395005 TI - N-cadherin regulates target specificity in the Drosophila visual system. AB - Using visual behavioral screens in Drosophila, we identified multiple alleles of N-cadherin. Removal of N-cadherin selectively from photoreceptor neurons (R cells) causes deficits in specific visual behaviors that correlate with disruptions in R cell connectivity. These defects include disruptions in the pattern of neuronal connections made by all three classes of R cells (R1-R6, R7, and R8). N-cadherin is expressed in both R cell axons and their targets. By inducing mitotic recombination in a subclass of eye progenitors, we generated mutant R7 axons surrounded by largely wild-type R cell axons and a wild-type target. R7 axons lacking N-cadherin mistarget to the R8 recipient layer. We consider the implications of these findings in the context of the proposed role for cadherins in target specificity. PMID- 11395006 TI - Na(+) action potentials in human photoreceptors. AB - Mammalian photoreceptors are hyperpolarized by a light stimulus and are commonly thought to be nonspiking neurons. We used the whole-cell patch-clamp technique on surgically excised human retina to examine whether human photoreceptors can elicit action potentials. We discovered that human rod photoreceptors express voltage-gated Na(+) channels, and generate Na(+) action potentials, in response to membrane depolarization from membrane potentials of -60 or -70 mV. Na(+) spikes in human rods were elicited at the termination of a light response that hyperpolarized the potential well below -50 mV. This served to amplify the release of a neurotransmitter when a bright light is turned off, and thus selectively amplify the off response to the light signal. PMID- 11395007 TI - Synaptotagmin VII as a plasma membrane Ca(2+) sensor in exocytosis. AB - Synaptotagmins I and II are Ca(2+) binding proteins of synaptic vesicles essential for fast Ca(2+)-triggered neurotransmitter release. However, central synapses and neuroendocrine cells lacking these synaptotagmins still exhibit Ca(2+)-evoked exocytosis. We now propose that synaptotagmin VII functions as a plasma membrane Ca(2+) sensor in synaptic exocytosis complementary to vesicular synaptotagmins. We show that alternatively spliced forms of synaptotagmin VII are expressed in a developmentally regulated pattern in brain and are concentrated in presynaptic active zones of central synapses. In neuroendocrine PC12 cells, the C(2)A and C(2)B domains of synaptotagmin VII are potent inhibitors of Ca(2+) dependent exocytosis, but only when they bind Ca(2+). Our data suggest that in synaptic vesicle exocytosis, distinct synaptotagmins function as independent Ca(2+) sensors on the two fusion partners, the plasma membrane (synaptotagmin VII) versus synaptic vesicles (synaptotagmins I and II). PMID- 11395008 TI - Drosophila Hsc70-4 is critical for neurotransmitter exocytosis in vivo. AB - Previous in vitro studies of cysteine-string protein (CSP) imply a potential role for the clathrin-uncoating ATPase Hsc70 in exocytosis. We show that hypomorphic mutations in Drosophila Hsc70-4 (Hsc4) impair nerve-evoked neurotransmitter release, but not synaptic vesicle recycling in vivo. The loss of release can be restored by increasing external or internal Ca(2+) and is caused by a reduced Ca(2+) sensitivity of exocytosis downstream of Ca(2+) entry. Hsc4 and CSP are likely to act in common pathways, as indicated by their in vitro protein interaction, the similar loss of evoked release in individual and double mutants, and genetic interactions causing a loss of release in trans-heterozygous hsc4-csp double mutants. We suggest that Hsc4 and CSP cooperatively augment the probability of release by increasing the Ca(2+) sensitivity of vesicle fusion. PMID- 11395009 TI - Dynamic visualization of local protein synthesis in hippocampal neurons. AB - Using pharmacological approaches, several recent studies suggest that local protein synthesis is required for synaptic plasticity. Convincing demonstrations of bona fide dendritic protein synthesis in mammalian neurons are rare, however. We developed a protein synthesis reporter in which the coding sequence of green fluorescent protein is flanked by the 5' and 3' untranslated regions from CAMKII alpha, conferring both dendritic mRNA localization and translational regulation. In cultured hippocampal neurons, we show that BDNF, a growth factor involved in synaptic plasticity, stimulates protein synthesis of the reporter in intact, mechanically, or "optically" isolated dendrites. The stimulation of protein synthesis is blocked by anisomycin and not observed in untreated neurons. In addition, dendrites appear to possess translational hot spots, regions near synapses where protein synthesis consistently occurs over time. PMID- 11395010 TI - A kainate receptor increases the efficacy of GABAergic synapses. AB - Brain functions are based on the dynamic interaction of excitatory and inhibitory inputs. Spillover of glutamate from excitatory synapses may diffuse to and modulate nearby inhibitory synapses. By recording unitary inhibitory postsynaptic currents (uIPSCs) from cell pairs in CA1 of the hippocampus, we demonstrated that low concentrations of Kainate receptor (KAR) agonists increased the success rate (P(s)) of uIPSCs, whereas high concentrations of KAR agonists depressed GABAergic synapses. Ambient glutamate released by basal activities or stimulation of the stratum radiatum increases the efficacy of GABAergic synapses by activating presynaptic KARs, which facilitate Ca(2+)-dependent GABA release. The results suggest that glutamate released from excitatory synapses may also function as an intermediary between excitatory and inhibitory synapses to protect overexcitation of local circuits. PMID- 11395011 TI - Disruption of KCC2 reveals an essential role of K-Cl cotransport already in early synaptic inhibition. AB - Synaptic inhibition by GABA(A) and glycine receptors, which are ligand-gated anion channels, depends on the electrochemical potential for chloride. Several potassium-chloride cotransporters can lower the intracellular chloride concentration [Cl(-)](i), including the neuronal isoform KCC2. We show that KCC2 knockout mice died immediately after birth due to severe motor deficits that also abolished respiration. Sciatic nerve recordings revealed abnormal spontaneous electrical activity and altered spinal cord responses to peripheral electrical stimuli. In the spinal cord of wild-type animals, the KCC2 protein was found at inhibitory synapses. Patch-clamp measurements of embryonic day 18.5 spinal cord motoneurons demonstrated an excitatory GABA and glycine action in the absence, but not in the presence, of KCC2, revealing a crucial role of KCC2 for synaptic inhibition. PMID- 11395012 TI - Differential functions of mPer1, mPer2, and mPer3 in the SCN circadian clock. AB - The role of mPer1 and mPer2 in regulating circadian rhythms was assessed by disrupting these genes. Mice homozygous for the targeted allele of either mPer1 or mPer2 had severely disrupted locomotor activity rhythms during extended exposure to constant darkness. Clock gene RNA rhythms were blunted in the suprachiasmatic nucleus of mPer2 mutant mice, but not of mPER1-deficient mice. Peak mPER and mCRY1 protein levels were reduced in both lines. Behavioral rhythms of mPer1/mPer3 and mPer2/mPer3 double-mutant mice resembled rhythms of mice with disruption of mPer1 or mPer2 alone, respectively, confirming the placement of mPer3 outside the core circadian clockwork. In contrast, mPer1/mPer2 double mutant mice were immediately arrhythmic. Thus, mPER1 influences rhythmicity primarily through interaction with other clock proteins, while mPER2 positively regulates rhythmic gene expression, and there is partial compensation between products of these two genes. PMID- 11395013 TI - Odor coding in the Drosophila antenna. AB - Odor coding in the Drosophila antenna is examined by a functional analysis of individual olfactory receptor neurons (ORNs) in vivo. Sixteen distinct classes of ORNs, each with a unique response spectrum to a panel of 47 diverse odors, are identified by extracellular recordings. ORNs exhibit multiple modes of response dynamics: an individual neuron can show either excitatory or inhibitory responses, and can exhibit different modes of termination kinetics, when stimulated with different odors. The 16 ORN classes are combined in stereotyped configurations within seven functional types of basiconic sensilla. One sensillum type contains four ORNs and the others contain two neurons, combined according to a strict pairing rule. We provide a functional map of ORNs, showing that each ORN class is restricted to a particular spatial domain on the antennal surface. PMID- 11395014 TI - Model of transient oscillatory synchronization in the locust antennal lobe. AB - Transient pairwise synchronization of locust antennal lobe (AL) projection neurons (PNs) occurs during odor responses. In a Hodgkin-Huxley-type model of the AL, interactions between excitatory PNs and inhibitory local neurons (LNs) created coherent network oscillations during odor stimulation. GABAergic interconnections between LNs led to competition among them such that different groups of LNs oscillated with periodic Ca(2+) spikes during different 50-250 ms temporal epochs, similar to those recorded in vivo. During these epochs, LN evoked IPSPs caused phase-locked, population oscillations in sets of postsynaptic PNs. The model shows how alternations of the inhibitory drive can temporally encode sensory information in networks of neurons without precisely tuned intrinsic oscillatory properties. PMID- 11395015 TI - Model of cellular and network mechanisms for odor-evoked temporal patterning in the locust antennal lobe. AB - Locust antennal lobe (AL) projection neurons (PNs) respond to olfactory stimuli with sequences of depolarizing and hyperpolarizing epochs, each lasting hundreds of milliseconds. A computer simulation of an AL network was used to test the hypothesis that slow inhibitory connections between local neurons (LNs) and PNs are responsible for temporal patterning. Activation of slow inhibitory receptors on PNs by the same GABAergic synapses that underlie fast oscillatory synchronization of PNs was sufficient to shape slow response modulations. This slow stimulus- and neuron-specific patterning of AL activity was resistant to blockade of fast inhibition. Fast and slow inhibitory mechanisms at synapses between LNs and PNs can thus form dynamical PN assemblies whose elements synchronize transiently and oscillate collectively, as observed not only in the locust AL, but also in the vertebrate olfactory bulb. PMID- 11395016 TI - Search efficiency but not response interference affects visual selection in frontal eye field. AB - Two manipulations of a visual search task were used to test the hypothesis that the discrimination of a target from distractors by visually responsive neurons in the frontal eye field (FEF) marks the outcome and conclusion of visual processing instead of saccade preparation. First, search efficiency was reduced by increasing the similarity of the distractors to the target. Second, response interference was introduced by infrequently changing the location of the target in the array. Both manipulations increased reaction time, but only the change in search efficiency affected the time needed to select the target by visually responsive neurons. This result indicates that visually responsive neurons in FEF form an explicit representation of the location of the target in the image. PMID- 11395017 TI - Neuronal correlates of motor performance and motor learning in the primary motor cortex of monkeys adapting to an external force field. AB - The primary motor cortex (M1) is known to control motor performance. Recent findings have also implicated M1 in motor learning, as neurons in this area show learning-related plasticity. In the present study, we analyzed the neuronal activity recorded in M1 in a force field adaptation task. Our goal was to investigate the neuronal reorganization across behavioral epochs (before, during, and after adaptation). Here we report two main findings. First, memory cells were present in two classes. With respect to the changes of preferred direction (Pd), these two classes complemented each other after readaptation. Second, for the entire neuronal population, the shift of Pd matched the shift observed for muscles. These results provide a framework whereby the activity of distinct neuronal subpopulations combines to subserve both functions of motor performance and motor learning. PMID- 11395018 TI - Functional interactions of the inferior frontal cortex during the processing of words and word-like stimuli. AB - The hypothesis that ventral/anterior left inferior frontal gyrus (LIFG) subserves semantic processing and dorsal/posterior LIFG subserves phonological processing was tested by determining the pattern of functional connectivity of these regions with regions in left occipital and temporal cortex during the processing of words and word-like stimuli. In accordance with the hypothesis, we found strong functional connectivity between activity in ventral LIFG and activity in occipital and temporal cortex only for words, and strong functional connectivity between activity in dorsal LIFG and activity in occipital and temporal cortex for words, pseudowords, and letter strings, but not for false font strings. These results demonstrate a task-dependent functional fractionation of the LIFG in terms of its functional links with posterior brain areas. PMID- 11395019 TI - Functional imaging of neural responses to expectancy and experience of monetary gains and losses. AB - Neural responses accompanying anticipation and experience of monetary gains and losses were monitored by functional magnetic resonance imaging. Trials comprised an initial "prospect" (expectancy) phase, when a set of three monetary amounts was displayed, and a subsequent "outcome" phase, when one of these amounts was awarded. Hemodynamic responses in the sublenticular extended amygdala (SLEA) and orbital gyrus tracked the expected values of the prospects, and responses to the highest value set of outcomes increased monotonically with monetary value in the nucleus accumbens, SLEA, and hypothalamus. Responses to prospects and outcomes were generally, but not always, seen in the same regions. The overlap of the observed activations with those seen previously in response to tactile stimuli, gustatory stimuli, and euphoria-inducing drugs is consistent with a contribution of common circuitry to the processing of diverse rewards. PMID- 11395020 TI - Insulin resistance in the St. Thomas' mixed hyperlipidaemic (SMHL) rabbit, a model for familial combined hyperlipidaemia. AB - The St. Thomas mixed hyperlipidaemic (SMHL) rabbit exhibits an inherited hyperlipidaemia similar to that seen in familial combined hyperlipidaemia (FCHL). In this study, we investigated whether the SMHL rabbit is insulin resistant, a condition often associated with FCHL. Six young and six mature combined hyperlipidaemic SMHL rabbits, age/sex matched New Zealand White (NZW) control rabbits and six young hypercholesterolaemic Watanabe heritable hyperlipidaemic (WHHL) control rabbits were fed a 0.08% (w/w) cholesterol-enriched diet for at least 1 month prior to the start of the experiment. We performed an oral glucose tolerance test after an overnight fast by dosing the rabbits with a solution of 1 g of glucose per kg body weight. Blood was withdrawn just before and 15, 30, 45, 60 and 120 min after administration of the oral glucose dose. Plasma glucose levels were similar in SMHL, WHHL and NZW rabbits throughout the oral glucose tolerance test. Fasting glucose levels were slightly increased in WHHL rabbits but not in young and adult SMHL rabbits as compared to NZW rabbits. The area under the curve (AUC) for the insulin response was significantly increased for both young (P<0.05) and mature (P<0.05) SMHL rabbits, and in WHHL rabbits, compared with NZW rabbits. The AUC for the ratio of glucose:insulin response to the glucose dose was decreased in young and mature SMHL rabbits (P<0.05 and P<0.01, respectively) and in young WHHL rabbits (P<0.05), compared with NZW rabbits. Only WHHL rabbits showed an increased AUC for the non-esterified fatty acid response compared to NZW rabbits. Log-transformed plasma triglycerides values were significantly correlated with the log-transformed AUC for the insulin response in young SMHL rabbits (r=0.81; P<0.05) and with the AUC for the insulin response in mature SMHL rabbits (r=0.84; P<0.05). WHHL rabbits showed no significant correlation. In conclusion, SMHL rabbits are insulin resistant, the severity of which appears to increase with age. Therefore, the SMHL rabbit offers a valuable animal model in which to study the relation between hypertriglyceridaemia and insulin resistance. PMID- 11395021 TI - Effects of TCV-116 on expression of NOS and adrenomedullin in failing heart of Dahl salt-sensitive rats. AB - We examined the effects of TCV-116, an angiotensin II type 1 receptor antagonist, on endothelial-cell nitric oxide synthase (eNOS), inducible NOS (iNOS), and adrenomedullin (ADM) expression in the left ventricle (LV) and evaluated these relation to myocardial remodeling in failing heart of Dahl salt-sensitive hypertensive rats (DS) fed a high-salt diet. TCV-116 (DSHF-T, 5 mg/kg/day, subdepressor dose) or vehicle (DSHF-V) were given from left ventricular hypertrophy to heart failure stage for 7 weeks. Markedly increased left ventricular end-diastolic diameter and reduced fractional shortening in DSHF-V was significantly ameliorated in DSHF-T. The eNOS mRNA and protein in the LV was significantly suppressed in DSHF-V compared with control rats (DR-C), and significantly increased in DSHF-T compared with DSHF-V. The iNOS mRNA and protein, ADM mRNA and immunoreactive ADM contents, and type I collagen mRNA in the LV were significantly increased in DSHF-V compared with DR-C, and significantly decreased in DSHF-T compared with DSHF-V. DSHF-V showed a significant increase of the wall-to-lumen ratio, perivascular fibrosis, and myocardial fibrosis, with all these parameters being significantly improved by TCV-116. In conclusion, myocardial remodeling and heart failure in DS rats fed a high-salt diet were significantly ameliorated by a subdepressor dose of TCV-116, which may be due to a increased in eNOS and a decreased in iNOS mRNA and protein expression in the LV. Moreover, the ADM mRNA and immunoreactive ADM contents are upregulated in failing heart of DS rats fed a high-salt diet, and increased ADM expression may have a role in the defense mechanism against further cardiac dysfunction and impaired myocardial remodeling. PMID- 11395022 TI - Expression of PDGF receptors and ligand-induced migration of partially differentiated human monocyte-derived macrophages. Influence of IFN-gamma and TGF beta. AB - In the early atherosclerotic lesion, monocytes accumulate at sites of inflammation and endothelial injury. Platelet-derived growth factor (PDGF), produced for example by macrophages, is a chemoattractant for smooth muscle cells and possibly also for macrophages. During early differentiation into macrophages, human monocytes (early hMDM) showed lower expression of PDGF alpha-receptor (PDGF Ralpha) than beta-receptor (PDGF-Rbeta) mRNA. Early hMDM showed increased random motility (chemokinesis) in the presence of PDGF of the long (BB(L)) but not short (BB(S)) B-chain homodimer. Neither PDGF-AA(S) nor PDGF-AA(L) affected early hMDM motility. Since increased cytokine levels accompany inflammation, the influence of interferon-gamma (IFN-gamma) and transforming growth factor-beta (TGF-beta) on PDGF-R expression and migratory response were studied. Only PDGF-Ralpha mRNA was highly upregulated by IFN-gamma. TGF-beta only had minor effects on receptor mRNAs. Upregulation of PDGF-Ralpha levels by IFN-gamma was accompanied by significantly increased migration (chemotaxis) towards PDGF-AA(L) only. Consequently, IFN-gamma modulates PDGF-Rs expression in early hMDM and, subsequently, the chemotactic activity of PDGF-AA(L) on IFN-gamma-stimulated early hMDM. This suggests that PDGF-AA(L) may be involved in attracting activated monocytes to sites of inflammation and injury. PMID- 11395023 TI - Development of a lipid-rich, soft plaque in rabbits, monitored by histology and intravascular ultrasound. AB - Lipid rich, soft plaques in the clinic are a common forerunner to occlusive thrombus formation, even with modest arterial stenosis. Animal models of atherosclerosis, obtained by various methods, do not generally allow direct in vivo evaluation of the lesion and, furthermore, cannot be examined more than once. The aim of the study was the generation of a rabbit model of atherosclerosis, with morphological characteristics similar to human lipid-rich, soft atheromatous plaques, and the evaluation of the reliability of intravascular ultrasound (IVUS) technology in the study of the development of atherosclerotic lesions in this model. Briefly, New Zealand white rabbits undergo perivascular electrical injury at both common carotid arteries, together with a 1.5% cholesterol diet for up to 90 days. The lesioned arterial segments show progressive changes, from diffuse cellular mortality, to macrophage infiltration in the media, up to the final migration of macrophages to the neointima, resulting in bulky, eccentric, macrophage and lipid-rich lesions. At IVUS, the produced lesions clearly resemble those described as 'soft plaques' in the clinical setting, with minimal calcification and reduced echo-reflectivity versus the adventitial layer. Quantitative and morphometric analysis of plaques shows a significant correlation between histological and IVUS measurements at each time point. In conclusion, vascular injury in the common carotids of rabbits generates atherosclerotic lipid-rich, soft plaques, that can be properly assessed by the IVUS methodology. The easy accessibility of the arterial lesion allows serial IVUS investigations and the direct evaluation of a number of locally or generally delivered therapeutic agents. PMID- 11395024 TI - Downregulation of the ERK 1 and 2 mitogen activated protein kinases using antisense oligonucleotides inhibits proliferation of porcine vascular smooth muscle cells. AB - The current model of the arterial response to injury suggests that proliferation of vascular smooth muscle cells is a central event. Mitogen activated protein kinases are part of the final common pathway of intracellular signalling involved in cell division and thus constitute an attractive target in attempting to inhibit this proliferation. We hypothesised that antisense oligonucleotides to mitogen activated protein kinase would inhibit serum induced smooth muscle cell proliferation by downregulating the protein. Porcine vascular smooth muscle cells were cultured and an antisense oligonucleotide sequence against the ERK family of mitogen activated protein kinases (AMK1) was introduced by liposomal transfection. Sense oligonucleotides and a random sequence were used as controls. Proliferation was inhibited by AMK1 versus the sense controls, as assessed by tritiated thymidine incorporation (P<0.01). Immunoblots revealed downregulation of the target protein by AMK1 by 63% versus the sense control (P<0.05). In conclusion, antisense oligonucleotides specifically inhibited proliferation and downregulated the target protein. This is consistent with a central role for mitogen activated protein kinases in vascular smooth muscle cell proliferation in the porcine model. In addition, the data suggest a possible role for antisense oligonucleotides in the modulation of the arterial injury response. PMID- 11395025 TI - Localization of CD36 and scavenger receptor class A in human coronary arteries--a possible difference in the contribution of both receptors to plaque formation. AB - CD36 and scavenger receptor class A types I and II (SR-AI/II) are major receptors for oxidized low density lipoproteins (OxLDL) expressed on macrophages. To elucidate the role of these two macrophage scavenger receptors in the development of coronary atherosclerosis, we examined the localization of CD36 and SR-AI/II in human coronary atherosclerotic lesions. Serial cryostat sections of 49 coronary arteries obtained from 43 autopsied cases were examined immunohistochemically. Regarding the relationship between the severity of atherosclerosis and immunoreactivities to CD36, there was almost no immunoreactivity to CD36 in regions with diffuse intimal thickening, while the expression of CD36 was accelerated in parallel with the progression of atherosclerosis. In contrast, SR AI/II was expressed persistently from regions with diffuse intimal thickening to atherosclerotic plaques. We also clarified the differential distribution of CD36 and SR-AI/II in atheromatous plaques. Close to the luminal surface of the intima, macrophages were relatively small in size, contained lesser lipids, and expressed SR-AI/II more abundantly than CD36. In contrast, macrophages in the core region were larger in size, contained more lipids, were strongly positive for CD36 and showed a weaker immunoreactivity to SR-AI/II than those in the luminal surface of the intima. In conclusion, the expression of CD36 and SR-AI/II on macrophages may be regulated differently in the process of coronary atherogenesis. PMID- 11395026 TI - Effect of atherogenic diet on reverse cholesterol transport in vivo in atherosclerosis susceptible (C57BL/6) and resistant (C3H) mice. AB - Mice susceptible (C57BL/6) or resistant (C3H) to atherosclerosis induced by a high cholesterol-cholate containing diet (A-diet) were used to study reverse cholesterol transport (RCT) in vivo as measured by loss of cholesterol from a depot created by injection of cationized LDL into the rectus femoris muscle. Plasma total and HDL-cholesterol (HDL-C), total and HDL phospholipid (HDL-PL) levels in chow fed C3H male and female mice were higher than in C57BL/6 mice. After one month on A-diet, plasma cholesterol more than doubled in both strains and genders. The decrease in HDL-C and HDL-PL was twice as great in C57BL/6 as in C3H female mice, while in male C3H mice there was no decrease. The loss of exogenous cholesterol mass (ECM) after injection of cationized LDL was more rapid in C3H than in C57BL/6 mice. In chow fed mice, ECM retained in muscle on day 12 was 37% in C57BL/6 and 20% in C3H females; in males it was 39% and 18% in C57BL/6 and C3H, respectively. On A-diet, 76% were retained in C57BL/6 and 28% in C3H females; these values were 59% and 28% in C57BL/6 and C3H males. Thus, the slow clearance of ECM (which represents RCT) in C57BL/6 mice on A-diet, that could be related to a marked decrease of HDL-PL, might contribute towards their susceptibility to atherosclerosis. PMID- 11395027 TI - Involvement of interleukin-6 in atherosclerosis but not in the prevention of fatty streak formation by 17beta-estradiol in apolipoprotein E-deficient mice. AB - Interleukin-6 (IL-6) gene expressed in bone marrow-derived stromal cells and osteoblasts contributes to the state of mineralization and its control by estradiol may be involved in the development of post-menopausal osteoporosis. Since IL-6 is also expressed in the different cell populations of the arterial wall, the purpose of this study was to gain more insight into its involvement in the atherosclerotic process and the atheroprotective effect of estradiol by studying double deficient mice at the apolipoprotein E and IL-6 loci (IL-6(-/ )/E(-/-)). At 1 year of age, IL-6(-/-)/E(-/-) mice showed similar hypercholesterolemia to IL-6(+/+)/E(-/-) mice but presented significantly larger and more calcified lesions. In younger mice (sixteen weeks of age), no significant difference in fatty streaks could be detected in IL-6(+/+)/E(-/-), IL 6(+/-)/E(-/-) and IL-6(-/-)/E(-/-) mice on a normal chow diet. Estrogen supplementation at this age induced a decrease of fatty streak formation in all three genotypes. The combined data indicate that IL-6 expression is involved at the fibrous plaque stage of the atherosclerotic process but does not constitute a direct target for estradiol to prevent fatty streak formation. PMID- 11395028 TI - Interaction of transmural pressure and shear stress in the transport of albumin across the rabbit aortic wall. AB - In this study the synergistic role of the two haemodynamic parameters, pressure and wall shear stress, in macromolecular transport has been examined across the wall of the rabbit thoracic aorta. Arteries were subjected to 70 and 150 cm water pressure in the presence of fluid flow imposed shear stress. The flux of FITC labelled bovine serum albumin was found to be 3.36+/-1.34 x 10(-6) and 1.99+/ 0.77 x 10(-6) cm/s (mean+/-S.D.) after 90 min incubation at 70 and 150 cm water, respectively. The mean relative tissue concentrations were 0.0039+/-0.0025 and 0.012+/-0.007 at 70 and 150 cm water, respectively. Under low values of steady wall shear stress, efflux of BSA is retarded at 150 cm since its tissue concentration is found to be higher than at 70 cm. The net outcome arises as a result of the interaction of increased permeability of endothelial cells exposed to shear stress, the pressure induced distension of the wall matrix and the differential effect of EDRF/NO at the two pressures on medial hydraulic conductivity. In the presence of the EDRF/NO inhibitor L-NAME, reduction in flux of albumin was observed at both the pressures, the decrease being greater at 150 cm water. In the absence of EDRF, the NO synthase independent vasodilator EDHF may be released, which maintains the tone of the medial smooth muscle low. Action of EDHF may be more marked at 150 cm water because NO synthesis is attenuated by higher transmural pressure and the presence of L-NAME eliminates shear stress stimulated NO release. Consequently the dilated vessel will have decreased porosity and less albumin space. The BSA flux across the aorta is, therefore, influenced by both endothelial permeability and permeability of the medial matrix, which are in turn modulated by an interplay of transmural pressure and fluid flow generated shear stress. PMID- 11395029 TI - Randomised controlled trial of use by hypercholesterolaemic patients of a vegetable oil sterol-enriched fat spread. AB - Plant sterols may be a useful additive therapy in the treatment of hypercholesterolaemic patients. The purpose of this study was to determine the effect of a fat spread enriched with vegetable oil sterols on plasma lipid, lipoprotein and apolipoprotein concentrations. A randomised double blind placebo controlled crossover trial with two consecutive periods of 8 weeks was conducted. 30 patients with heterozygous familial hypercholesterolaemia treated concurrently with an HMG-CoA reductase inhibitor (statin) and 32 patients with type IIa primary hypercholesterolaemia with a total cholesterol concentration >6.5 mmol/l not taking lipid-lowering drug therapy were recruited from a hospital lipid clinic. The active treatment was a fortified fat spread (25 g/day) providing 2.5 g of plant sterols. The control spread was indistinguishable in taste and appearance. Comparison at the end of the two 8-week trial periods showed a statistically significant reduction in total and LDL-cholesterol with use of the fortified spread but the results were confounded by a carry-over effect, which was partly explained by changes in the background diet. Because a carry-over effect was present, further analyses were restricted to the parallel arms of the first treatment period and were conducted on an intention to treat basis. After 4 weeks, LDL-cholesterol had decreased by 0.04 mmol/l ([0.8%] 95% confidence interval -0.44-0.37 NS) in the placebo group and decreased by -0.76 mmol/l ([15.0%] 95% CI -1.03--0.48, P<0.0001) in the active treatment group. After 8 weeks, the corresponding results were 0.0 mmol/l ([0.0%] 95% CI -0.26-0.24 NS) and -0.51 mmol/l ([10.0%] 95% CI -0.73--0.29 P<0.0001). There were no significant changes in apolipoprotein AI or B concentrations in the placebo group, but there was a small but statistically significant increase in apolipoprotein AI and a decrease in apolipoprotein B in the active treatment group. HDL cholesterol and triglyceride concentrations were unchanged. There was no difference in response between patients with statin-treated familial hypercholesterolaemia and patients with type IIa hyperlipoproteinaemia. We conclude that a fortified fat spread enriched with vegetable oil sterols reduces LDL-cholesterol by 10-15% with no difference in response between hypercholesterolaemic patients prescribed statins and those not taking lipid-lowering drug therapy. PMID- 11395030 TI - Transcriptional regulation of the human sterol 27-hydroxylase gene (CYP27) and promoter mapping. AB - Recent evidence suggests that sterol 27-hydroxylase may play a role in cholesterol homeostasis and affect atherogenesis. The major objective of the study was to map and characterize the sterol 27-hydroxylase (CYP27) promoter region. Here we show that CYP27 gene has a TATA-less promoter and transcription initiates at a cluster of sites. The basic promoter is located between -166 and 187 bp from the translation initiation site. Possible positive transcription regulation sites are located at position -187 to -320 and -857 to -1087 bp. A negative transcription regulator site is located in position -320 to -413 bp. An enhancer sequence is located upstream to position -1087. CYP27 is upregulated by dexamethasone and downregulated by cyclosporin A and cholic acid. The dexamethasone responsive element is located between 1087 and 678 bp upstream to the putative ATG. Cyclosporin A affects bile acid metabolism by repressing CYP27 at the transcriptional level. The cyclosporin A- responsive element is mapped to between 1087 and 4000 bp upstream of the ATG. Cholic acid represses sterol 27 hydroxylase mRNA level by affecting the stability of its mRNA. The results obtained here imply that CYP27 has a potentially important role in cholesterol homeostasis in human cells, and is regulated by several substances that were previously shown to affect bile acid metabolism. PMID- 11395031 TI - Endothelium-dependent vasodilation is related to the fatty acid composition of serum lipids in healthy subjects. AB - The fatty acid (FA) composition of the serum lipids has been associated with cardiovascular disease (CVD). As an attenuated endothelium-dependent vasodilation (EDV) has been suggested as an early marker of atherosclerosis, we investigated the relationships between the proportion of FA in serum lipids (cholesterol esters and phospholipids) together with the levels of serum LDL- and HDL cholesterol and triglycerides and EDV, as well as endothelium-independent vasodilation (EIDV). Fifty-six healthy subjects (31 men and 25 women), aged between 20 and 69 years, underwent measurements of forearm blood flow (FBF) at rest and during local infusion of 2 and 4 microg/min of metacholine (Mch, evaluating EDV), 5 and 10 microg/min of sodium nitroprusside (SNP, evaluating endothelium-independent vasodilation, EIDV) using venous occlusion plethysmography. An index of endothelial function was calculated as the ratio between EDV and EIDV. The proportion of palmitic (16:0) and palmitoleic (16:1) acids were inversely related (r=-0.35 and -0.35, P<0.01 for both), while linoleic acid (18:2 n6) and the HDL-cholesterol concentration were positively related (r=0.35 and 0.36, P<0.01 for both) to the endothelial function index. In multiple regression analysis also including age and gender, palmitoleic acid and HDL cholesterol were significant independent predictors of endothelial function. Alfa linolenic acid (18:3 n3) was positively correlated to both EDV and EIDV (r=0.40 and 0.43, P<0.01 for both), indicating a protective effect of this essential FA on vasodilation in general. It is concluded that the FA composition of serum lipids, partly reflecting the composition of dietary fat and previously associated with the development of CVD, was associated with endothelial function in apparently healthy subjects. PMID- 11395032 TI - Urinary mevalonate excretion rate in type 2 diabetes: role of metabolic control. AB - An increased cholesterogenesis has been described in obese dyslipidemic type 2 diabetic patients and in a small number of patients with poor glucose control. So far, it is not clear if increased cholesterogenesis in type 2 diabetes is related to the degree of glycemic control or depends on the commonly associated dyslipidemia or both. Therefore, the aim of the present study was to investigate the relationships among cholesterogenesis and degree of metabolic control in a group of non-obese normolipidemic type 2 diabetic patients. Fifty four (25 men and 29 postmenopausal women) non-obese type 2 diabetic patients with cholesterol and triglyceride plasma levels, respectively, below 6.40 and 2.85 mmol/l and 20 normal subjects matched for age and sex were studied. Endogenous cholesterol synthesis was evaluated by the determination of 24-h urinary mevalonate excretion (MVA). In the diabetic group the mean glycated hemoglobin was 8.47+/-2.2% (range 4.6-14.6%), the mean total cholesterol, triglycerides, HDL and LDL cholesterol were, respectively, 4.86+/-0.7, 1.64+/-0.5, 1.19+/-0.3 and 2.87+/-0.7 mmol/l. The mean 24-h MVA urine excretion rates were 1.41+/-0.3 micromol/24 h in control subjects and 1.66+/-0.7 micromol/24 h in diabetics (P=0.05). In diabetics, urinary mevalonate excretion was significantly correlated with glycated hemoglobin concentrations (HbA(1c)) (r=0.65; P=0.0001) and body mass index (BMI) (r=0.33; P=0.009). In the multivariate analysis both HbA(1c) and BMI were independent predictors of urinary mevalonate. These data demonstrate that lower the degree of blood glucose control, higher is the whole body cholesterol production even in the absence of overt dyslipidemia. In conclusion, the relationship between mevalonate excretion rate and glycated hemoglobin gives further weight to the importance of intensive blood-glucose control in diabetic disease and adds a new element to the list of potentially atherogenic factors strictly related to hyperglycemia in type 2 diabetic patients. PMID- 11395033 TI - Human apolipoprotein A-IV metabolism within triglyceride-rich lipoproteins and plasma. AB - In order to investigate the metabolism of apo A-IV within TRL and plasma, we assessed TRL and plasma apo A-IV kinetics in 19 and 4 subjects, respectively, consuming an average US diet for a 6-week period. At the end of this diet study, each subject received a primed-constant infusion of deuterated leucine over a 15 h time period with hourly feeding, and blood samples were drawn at 10 time points. TRL was separated by ultracentrifugation. Apo A-IV was isolated by immunoprecipitation and/or SDS-PAGE. Apo A-IV concentrations were determined by immunoelectrophoresis. Stable isotope tracer/tracee ratios were measured by gas chromatography/mass spectrometry, and the data were analyzed by multicompartmental modeling. The mean concentrations of plasma and TRL apo A-IV during the isotope infusion period were 21.0+/-3.2 and 0.66+/-0.25 mg/dl, respectively, and these values were 11.5 and 30.5% higher than those of fasting samples. The mean TRL and plasma apo A-IV residence times (RT) were 1.97+/-0.57 and 2.71+/-0.65 days, and transport rates (TR) were 0.17+/-0.19 and 3.90+/-1.24 mg/kg per day, respectively. There were significant correlations between TRL apo A-IV concentrations and TR (r(2)=0.79, P<0.001), and between TRL apo A-IV pool size and TRL cholesterol levels (r(2)=0.29, P=0.02). Our data indicated that; (1) TRL apo A-IV has a RT of 1.97 days which is similar to that earlier reported for HDL apo A-IV; (2) Apo A-IV recirculates between TRL and other slowly turning over pools; (3) the primary determinant of TRL apo A-IV levels is its TR; and (4) there is no correlation between TRL apo A-IV and apo B48 fractional catabolism in TRL. PMID- 11395034 TI - Smoking is associated with increased hepatic lipase activity, insulin resistance, dyslipidaemia and early atherosclerosis in Type 2 diabetes. AB - We have studied the relationships between hepatic lipase activity, smoking, dyslipidaemia insulin resistance, and early atherosclerosis in 67 Type 2 diabetic subjects, 47 non-smokers and 20 smokers. Insulin resistance was measured using an insulin modified frequently sampled intravenous glucose tolerance test. Early atherosclerosis was assessed using high-resolution ultrasound to measure carotid intima media thickness (IMT) and an arterial ultrasonic score (AUS). Smokers had higher serum cholesterol and triglyceride, lower HDL and HDL2 cholesterol as well as increased hepatic lipase activity. They were also more insulin resistant than non-smokers. Smokers also had higher patient AUS scores. On multiple regression analysis, hepatic lipase activity emerged as the most significant variable affecting patient AUS. We suggest that smoking accentuates the dyslipidaemia of Type 2 diabetic subjects and this is associated with increased hepatic lipase activity. This may be one mechanism whereby smoking further increases the risk of cardiovascular disease in Type 2 diabetes. PMID- 11395035 TI - Carotid and femoral ultrasound morphology screening and cardiovascular events in low risk subjects: a 10-year follow-up study (the CAFES-CAVE study(1)). AB - BACKGROUND: Subclinical arteriosclerotic lesions at the carotid and femoral bifurcations may be related to the occurrence of future cardiovascular events and of occult arteriosclerotic coronary disease. B-mode ultrasound of carotid and femoral arteriosclerotic bifurcation lesions may provide a simple screening method to select asymptomatic subjects at risk of future events. METHODS AND RESULTS: 13221 low-risk, healthy, asymptomatic individuals were included in a 10 year, prospective, follow-up based on carotid and femoral bifurcation morphology defined by B-mode ultrasound. Four classes were considered at inclusion (I: normal wall, II: wall thickening, III: non-stenosing plaques, IV: stenosing plaques). When 10000 subjects (75.6% of included subjects; 6055 males, 3945 females) completed the 10-year follow-up the study was concluded. At 10 years there were 10 events (out of 7989 subjects) in class I and 81 events in II (930 subjects; incidence=8.6%); 239 events were observed in class III (611 subjects; 39.28%) and 381 events (470 subjects; 81.06%) in IV; 61 deaths occurred in classes III+IV (1081 subjects) producing a death rate within these two classes of 5.5% (51 out of 61=81.5% in class IV). The increased event rates in classes III and IV were significant (log rank; P<0.02) in comparison with I and II. CONCLUSIONS: Carotid and femoral morphology identified 2011 subjects (20.1% of the population) in classes II,III,IV including 98.6% of cardiovascular events and deaths in the following 10 years. A higher (P<0.05) rate of progression in classes III and IV in comparison with I and II was also observed. The ultrasound carotid and femoral classification was useful in selecting subjects at very low risk of cardiovascular events (class I), those at limited risk (class II) and a group at moderate risk (class III). A subpopulation at high risk of cardiovascular events (IV) was identified. PMID- 11395036 TI - Histopathological modifications of early atherosclerotic lesions by risk factors- findings in PDAY subjects. AB - To investigate whether histopathological modifications on early atherosclerotic lesions differ according to risk factors, we compared the histological findings of arteries obtained from a multicenter study in the USA (Pathobiological Determinants of Atherosclerosis in Youth, PDAY) with the antemortem risk factors. The materials comprised aortas and left anterior descending (LAD) coronary arteries of 140 male subjects. Measurements of intimal thickness, classification of intimal lesions, and density of foam cells and intimal fibrosis at the determined sites of LAD and aorta were evaluated. In both arteries, intimal thickness of hypertensives was greater than the normotensives with no definite proliferation of foam cells. In aortas, hypercholesterolemia was associated with an increase in foam cells, but not with an increase in intimal thickness. HDL-C value correlated inversely with number of foam cells in both the arteries, and the degree of intimal thickness in LADs, where early appearance of advanced lesion such as preatheroma and atheroma, was also indicated in the low HDL-C group. Smokers had less number of foam cells in both the arteries and more intensive intimal fibrosis in LAD than non-smokers. Our study suggests that there are several ways to advanced atherosclerotic lesions by risk factors. PMID- 11395037 TI - Common mutations in the lipoprotein lipase gene (LPL): effects on HDL-cholesterol levels in a Chinese Canadian population. AB - BACKGROUND: favorable lipid profiles including low total serum cholesterol (TC), TC/HDL-cholesterol (HDL-C) ratio and elevated HDL-C levels have been previously reported in Chinese living in China. More recent data, however, suggests a changing trend toward decreased HDL-C and increased TC and LDL cholesterol (LDL C) in Chinese populations. Environmental factors likely contribute, in part, to these findings. However, genetic factors contributing to lipoprotein metabolism may also play a role in determining the lipid/lipoprotein phenotype observed in Chinese populations. Lipoprotein lipase (LPL) mutations have been associated with altered HDL-C concentrations in Caucasians but have not yet been studied in a large population of Chinese descent. METHODS: 1577 Chinese Canadians of Cantonese descent were recruited for a cardiovascular risk factor study. The frequency and effect of three LPL gene polymorphisms [Asp9Asn (D9N, n=374), Asn291Ser (N291S, n=321) and Ser447-Ter (S447X, n=403)] on serum HDL-C concentrations was assessed. All the three polymorphisms have been shown to alter HDL-C levels in different Caucasian populations. RESULTS: lower TC, LDL-C, and TG and higher HDL-C were observed in both male and female Chinese Canadian subjects compared to other population samples. The D9N and N291S LPL polymorphisms were identified in 1/374 (0.3%) and 5/321 (1.6%) subjects, respectively. Carrier frequency of the S447X mutation was (102/403) 25.3%. This S447X polymorphism was observed with higher frequency in males with HDL-C levels in the highest tertile compared with those in the lowest HDL-C tertile (carrier frequencies 37.3 vs. 19.4%) (P=0.046). CONCLUSION: in this cohort of Chinese Canadians, the serum lipid profiles were more favorable than what has been reported for Caucasian Canadians. A favorable spectrum of polymorphisms in the LPL gene may mitigate the adverse effects of western lifestyle on plasma lipoproteins in this cohort of Cantonese Canadians. PMID- 11395038 TI - The 1298A-->C polymorphism in methylenetetrahydrofolate reductase (MTHFR): in vitro expression and association with homocysteine. AB - A common mutation in methylenetetrahydrofolate reductase (MTHFR), 677C-->T, is associated with reduced enzyme activity, a thermolabile enzyme and mild hyperhomocysteinemia, a risk factor for vascular disease. Recently, a second common mutation (1298A-->C; glutamate to alanine) was reported, but this mutation was suggested to increase homocysteine only in individuals who carried the bp677 variant. To evaluate the functional consequences of this mutation, we performed site-directed mutagenesis and in vitro expression. For in vivo assessment of clinical impact, we examined the 1298A-->C genotypes and plasma homocysteine in 198 individuals from the NHLBI Family Heart Study that had previously been assessed for the 677 substitution. Site-directed mutagenesis of the human cDNA was performed to generate enzymes containing each of the two mutations, as well as an enzyme containing both substitutions. Enzyme activity and thermolability were assessed in bacterial extracts. The activity of the wild-type cDNA was designated as 100%; mutant enzymes containing the 1298 and 677 mutations separately had 68% (+/-5.0) and 45% (+/-10.8), respectively, of control activity while the enzyme containing both mutations had 41% (+/-12.8) of control activity. The 1298 mutation was not associated with a thermolabile enzyme. In the Family Heart Study, fasting homocysteine was significantly higher (P<0.05) in individuals heterozygous for both substitutions, compared to individuals who carried only the 677C-->T variant. This study suggests that two variants in MTHFR should be assessed as genetic risk factors for hyperhomocysteinemia. PMID- 11395039 TI - Soluble adhesion molecules and unstable coronary artery disease. AB - Leukocyte adhesion and transendothelial migration, prerequisites in the development of atherosclerosis, are largely mediated by adhesion molecules. In addition, unstable coronary syndromes usually involve platelet activation and thrombus formation at the site of atherosclerotic plaque. Therefore, we compared plasma levels of soluble P-selectin, a measurement of platelet activation, as well as E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) in patients with atherosclerosis undergoing coronary angiography (n=76). Soluble P-selectin levels, as measured by ELISA, were significantly elevated in patients with unstable (n=44) vs stable (n=32) atherosclerotic disease (73.0 +/- 2.5 ng/ml vs 52.3 +/- 3.0 ng/ml, respectively, P<0.01). By logistic regression analysis, plasma level of soluble P-selectin was an independent predictor of an unstable coronary syndrome (OR 4.2, CI 1.4-12.9, P<0.01). Soluble E-selectin level, a marker of endothelial activation, was associated with extent of atherosclerosis but did not correlate with disease stability. Interestingly, soluble P-selectin was inversely correlated with plasma levels of the antioxidant alpha-tocopherol (R=-0.443, P<0.001), a known inhibitor of platelet function. In summary, amongst the soluble adhesion molecules, only P selectin is significantly increased in patients with unstable coronary syndromes. This study suggests that platelet activation persists in patients with unstable coronary syndromes despite concurrent aspirin therapy. In addition, the beneficial effects of alpha-tocopherol in patients with cardiovascular disease may be related to inhibition of platelet function. PMID- 11395040 TI - Atherogenic lipoprotein changes in diabetic nephropathy. AB - Cardiovascular risk is increased in patients with diabetic nephropathy. The aim of this study was to examine the relative impacts of albuminuria and renal failure, the two important features of diabetic nephropathy, on potentially atherogenic lipoprotein changes in this condition. The subjects were 160 non diabetic healthy controls and a total of 200 type 2 diabetes patients with various degrees of nephropathy. The diabetic patients were divided into four groups by urinary albumin/creatinine ratio (U-ACR) and serum creatinine (S-Cr) levels: DM-1 (U-ACR< 30 mg/g, N=85), DM-2 (U-ACR=30-300 mg/g, N=48), DM-3 (U-ACR > 300 mg/g, N=29) and DM-4 (S-Cr>177 micromol/l or 2.0mg/dl, N=38). Lipids in very low (VLDL), intermediate (IDL), low (LDL), and high density (HDL) lipoproteins were measured following ultracentrifugation. VLDL-cholesterol (VLDL C) was elevated (by 73-100%) in diabetic patients and it did not differ among the stages of nephropathy. IDL-C was higher as the nephropathy stage was advanced, and the elevation was significant in the DM-3 (by 75%) and DM-4 (by 131%) groups. LDL-C was not elevated in diabetic patients and was not different among the stages of nephropathy. Reduction of HDL-C was significant in DM-1, DM-2 and DM-3 (by 12-16%) and it was more exaggerated in DM-4 (by 35%). Multiple regression analyses indicated that elevated S-Cr, but not U-ACR, was an independent factor associated with raised IDL-C and lowered HDL-C in diabetic patients. These results indicate that diabetic patients with nephropathy show multiple lipoprotein changes, and that renal failure has greater impact than albuminuria on abnormalities in IDL and HDL. These lipoprotein alterations may contribute to an increased cardiovascular risk in diabetic nephropathy, especially in diabetic renal failure. PMID- 11395041 TI - Measures of oxidative stress in heterozygous familial hypercholesterolaemia. AB - Familial hypercholesterolaemia (FH) may be associated with increased oxidative stress which may contribute to atherogenesis. Plasma lipid hydroperoxides (ROOHs), 8-epi PGF(2alpha) and alpha-tocopherol were measured in normal subjects and in newly referred heterozygous FH patients and used as indices of oxidative stress. ROOH levels were higher (+16%), albeit non-significantly, in FH patients than in controls subjects (4.4+/-0.3 vs. 3.8+/-0.3 micromol/l; n=51 and 40, respectively). 8-epi PGF(2alpha) levels were significantly greater (+56%) in the FH patients than in controls (0.43+/-0.06 vs. 0.27+/-0.05 nmol/l; P<0.05; n=14 and 16, respectively). FH patients with vascular disease had significantly higher (+32%) levels of ROOH compared with patients without vascular disease (4.9+/-0.40 vs. 3.7+/-0.33 micromol/l; P<0.05; n=27 and 24, respectively). Similarly, 8-epi PGF(2alpha) concentrations were higher (+100%) in the FH patients with vascular disease than in those without it (0.6+/-0.08 vs. 0.3+/-0.10 nmol/l; P<0.05; n=6 and 8, respectively). Absolute alpha-tocopherol levels in FH patients were similar to those in controls (21.0+/-0.70 vs. 23.8+/-1.30 micromol/l). When alpha tocopherol levels were expressed relative to cholesterol, however, the concentrations were found to be significantly lower (-43%) in FH patients than in controls (2.9+/-0.10 vs. 5.1+/-0.40 micromol/mmol, P<0.0005). There were no differences in absolute or cholesterol standardised alpha-tocopherol levels in patients with and without vascular disease. These data suggest that oxidative stress is increased in FH-patients and is particularly pronounced in those patients with vascular disease. It is possible that increased oxidative stress may precede the development of vascular disease. PMID- 11395042 TI - Relationship of age-related myocardial infarction risk and Gln/Arg 192 variants of the human paraoxonase1 gene: the REGICOR study. AB - Paraoxonase1 (PON1) seems to exert a major antioxidant effect by removing lipid peroxidation products. A common polymorphism of the PON1 gene, the PON1-192 genetic polymorphism, modulates PON1 activity and has been related in some studies to coronary heart disease. Oxidized LDL is believed to play a crucial role in the pathogenesis of atherosclerosis and there are studies providing support for the oxidative stress theory of aging. We have conducted a case control study to determine whether PON1 activity and PON1-192 genetic variants have a different impact on myocardial infarction (MI) risk among individuals stratified by tertiles of age distribution. PON1-192 genotypes and PON1 activity were determined in 280 consecutive MI patients and 396 control subjects. Serum PON1 activity levels were significantly higher in controls than in MI patients [226 U/l (159-351) vs. 216 U/l (146-298), median (interquartile range), P=0.005]. A decline of PON1 activity levels with advancing age in subjects carrying the low activity QQ genotype was observed, particularly in MI patients. PON1 activity and age negatively correlated in MI patients but not in controls. In the entire population, middle-aged and older subjects showed MI risks of 1.89 (P=0.012) and 2.69 (P<0.001) respectively, compared with young subjects. These risks increased to 2.41 (P=0.016) and 4.39 (P<0.001), respectively, in QQ homozygotes in comparison with younger QQ homozygotes, decreased to 1.53 (P=0.314) and 2.08 (P=0.112), respectively, in QR heterozygotes, and also lowered to 1.95 (P=0.410) and 0.51 (P=0.508) in RR homozygotes who were middle-aged and older, respectively, compared with younger RR carriers. The effect of PON1-192 genotypes on the association of the older age-category and MI risk was gene-dosage related. PON1 activity decreases as a function of age in subjects homozygous for the Q allele. Age may also act on MI risk as a function of PON1-192 alleles. The risk of MI increases with advancing age, principally among subjects carrying the low activity QQ genotype. PMID- 11395043 TI - The association of c-reactive protein, serum amyloid a and fibrinogen with prevalent coronary heart disease--baseline findings of the PAIS project. AB - Recent data suggest that infections, inflammation and the immune system are involved in the process of atherosclerosis. The aim of the present study was to analyze the association of coronary heart disease (CHD) with three inflammation markers, C-reactive protein (CRP), serum amyloid-A (SAA) and plasma fibrinogen. The cross-sectional study included 1400 men aged 45-74 years, who participated in a cardiovascular risk factor survey in Finland in 1997. Participants with prevalent CHD had markedly higher CRP, SAA and fibrinogen levels than participants without CHD. In logistic regression models, the age, smoking, serum cholesterol and systolic blood pressure adjusted odds ratios (2nd, 3rd and 4th quartile as compared with the 1st quartile) of CHD increased gradually with increasing quartile of CRP (1.90, 2.27, 2.64), SAA (1.68, 1.83, 2.41), and fibrinogen (1.60, 1.95, 2.14). The associations weakened somewhat after further adjustment for indicators of obesity, particularly waist hip-ratio. CRP, SAA and fibrinogen levels were markedly lower among CHD patients using cholesterol lowering medication as compared to non-users. In conclusion, CRP, SAA and fibrinogen, which are markers of inflammation, were positively and significantly associated with prevalent CHD. Central obesity needs to be considered as a confounding factor in the observed associations. These findings support the hypothesis that cholesterol-lowering drugs have an anti-inflammatory effect. PMID- 11395044 TI - Impaired fibrinolysis and increased fibrinogen levels in South Asian subjects. AB - The potential role of haemostatic risk markers is largely unexplored in South Asians, who have increased morbidity and mortality from cardiovascular disease and an increased prevalence of insulin resistance. To investigate differences in thrombotic risk markers between South Asian and White populations, 42 Asian and 50 White males and 96 Asian and 80 White females, clinically free from vascular disease, were recruited. Venous blood samples were taken for measures of haemostasis and determination of blood lipids. South Asian females showed lower fasting blood glucose than White females (4.6 vs. 4.8 mmol/l, P<0.008). In the South Asian population, total cholesterol was lower in females, with a similar trend in males (females 5.0 vs. 5.5 mmol/l, P<0.001; males 5.1 vs. WM 5.5 mmol/l, P=0.09), but no difference in triglyceride levels. South Asian subjects of both genders had markedly higher levels of fibrinogen (females 3.3 vs. 2.8 mg/dl, P<0.0005; males 3.0 vs. 2.5 mg/dl P<0.002) and PAI-1 activity (females 14.6 vs. 8.7 ng/ml, P<0.0005, males 21.3 vs. 12.2 ng/ml, ) P<0.0005). Factor VII:C was lower in both South Asian groups (females 110.9 vs. 122.4%, P<0.005; males 103.3 vs. 125%, P<0.0005). Factor XII was lower in South Asian females and there were no differences in Factor XII levels in male populations. These results suggest that elevated PAI-1 and fibrinogen in Asians of both genders may contribute to the increased vascular risk experienced in this population; however, the role of dyslipidaemia and Factor VII are not clear in these processes. PMID- 11395045 TI - Glycoprotein Ia C807T polymorphism and risk of restenosis following coronary stenting. AB - Platelets are thought to contribute to development of restenosis following percutaneous coronary interventions. The glycoprotein Ia/IIa complex is a major platelet collagen receptor, its surface expression being influenced by two, linked single nucleotide polymorphisms (C807T and G873A) in the glycoprotein Ia gene. T807 is associated with increased expression of this integrin receptor. We assessed whether T807 is associated with an increased risk of restenosis in 1769 consecutive patients treated with coronary stenting. 6-month follow-up angiograms were available in 82.4% of the patients. C807T genotype distribution was CC in 35.8%, CT in 47.6% and TT in 16.6% of the patients. Restenosis (diameter stenosis > or =50% at follow-up angiography) occurred in 32.9% of CC, 31.5% of CT and 32.1% of TT patients (P=0.87). The rate of major adverse cardiac events (death, myocardial infarction or need of reintervention) within 1 yr was 21.6% for CC, 21.7% for CT and 21.2% for TT patients (P=0.98). Thus, carriage of the GP Ia T807 allele is not associated with an increased risk of restenosis or unfavorable late outcome following coronary artery stenting. PMID- 11395046 TI - Reduced myocardial flow reserve relates to increased carotid intima-media thickness in healthy young men. AB - Increased carotid artery wall thickness and lipoprotein oxidation are key early events in atherosclerosis. To test the hypothesis that reduced myocardial flow reserve is a marker of subclinical atherosclerosis, we examined the relationships between flow reserve and carotid artery intima-media thickness (IMT) in young men free from coronary heart disease. Basal and dipyridamole stimulated coronary blood flow was measured using positron emission tomography (PET) in 55 healthy men aged 36+/-4 years. Myocardial flow reserve was calculated as the ratio of stimulated flow to basal flow. The mean carotid artery IMT was measured using high-resolution ultrasound. Oxidised LDL was measured as baseline LDL diene conjugation. Myocardial flow reserve decreased across the quartiles of increasing IMT (P=0.006), and was 5.2+/-1.9 in the lowest quartile for IMT and 3.7+/-1.2 in the highest (P=0.04, I vs. IV quartile). In univariate analysis, oxidised LDL correlated inversely with flow reserve (r=-0.35, P=0.01) and directly with IMT (r=0.51, P<0.001). The association between flow reserve and IMT remained significant (P< or =0.01) in multivariate regression model including age, blood pressure, left ventricular mass, ox-LDL, total cholesterol, HDL-cholesterol and triglycerides as covariates. These data support the concept that reduced myocardial flow reserve reflects subclinical atherosclerosis in asymptomatic subjects, and suggest that increased lipoprotein oxidation is directly related to early structural and functional atherosclerotic vascular changes. PMID- 11395047 TI - Modeling cancer detection: tumor size as a source of information on unobservable stages of carcinogenesis. AB - This paper is concerned with modern approaches to mechanistic modeling of the process of cancer detection. Measurements of tumor size at diagnosis represent a valuable source of information to enrich statistical inference on the processes underlying tumor latency. One possible way of utilizing this information is to model cancer detection as a quantal response variable. In doing so, one relates the chance of detecting a tumor to its current size. We present various theoretical results emerging from this approach and illustrate their usefulness with numerical examples and analyses of epidemiological data. An alternative approach based on a threshold type mechanism of tumor detection is briefly described. PMID- 11395048 TI - Models for transmission of disease with immigration of infectives. AB - Simple models for disease transmission that include immigration of infective individuals and variable population size are constructed and analyzed. A model with a general contact rate for a disease that confers no immunity admits a unique endemic equilibrium that is globally stable. A model with mass action incidence for a disease in which infectives either die or recover with permanent immunity has the same qualitative behavior. This latter result is proved by reducing the system to an integro-differential equation. If mass action incidence is replaced by a general contact rate, then the same result is proved locally for a disease that causes fatalities. Threshold-like results are given, but in the presence of immigration of infectives there is no disease-free equilibrium. A considerable reduction of infectives is suggested by the incorporation of screening and quarantining of infectives in a model for HIV transmission in a prison system. PMID- 11395049 TI - A mathematical algorithm that computes breast cancer sizes and doubling times detected by screening. AB - This paper presents a mathematical algorithm that computes the sizes and growth rates of breast cancer detected in a hypothetical population that is screened for the disease. The algorithm works by simulating the outcomes of the hypothetical population twice, first without screening and then with screening. The simulation without screening relies on an underlying model of the natural history of the disease. The simulation with screening uses this natural history model to track the growth of breast tumors backwards in the time starting from the time they would have been detected without screening. The method of tracking tumor growth backward in time is different from methods that track tumor growth forward in time by starting from an estimated time of tumor onset. The screening algorithm combines the natural history model, the method tracking of tumor growth backward in time, the age group, the interval between screening exams, and the detection threshold of the screening exam to compute the joint distribution of tumor size and growth rate among screen-detected and interval patients. The algorithm also computes the sensitivity and leadtime distribution. It allows for arbitrary age groups, detection thresholds and screening intervals and may contribute to the design of future screening trials. PMID- 11395124 TI - Importance of hyperhomocysteinemia as a risk factor for venous thromboembolism in a Taiwanese population. A case-control study. AB - OBJECTIVE: To determine the current status of hyperhomocysteinemia, which is a known risk for venous thrombosis (DVT), in Taiwan. SUBJECTS: 101 unselected patients with a minimum of one episode of deep leg DVT, either initial inpatients or current compliant outpatients in a teaching hospital. METHODS: Various thrombophilic risks, gene polymorphism and clinical predisposition were evaluated. RESULTS AND CONCLUSIONS: Patients presented higher fast total plasma homocysteine (hcy) levels than age- and sex-matched controls did (14.1 vs. 9.94 microM). Based on the 95th percentile of control values, hyperhomocysteinemia had a four- to nine-fold risk for DVT, irrespective of clinical predisposition, as well as other thrombophilic risks surveyed. Polymorphism of a metabolizing enzyme, methylenetetrahydrofolate reductase (MTHFR), was not associated with DVT, although homozygous thermolabile mutation tended to have higher plasma hcy levels. Factor V Leiden was absent in analysis of 80 patients. In complete evaluation (hcy, antithrombin (AT), protein S (PS), protein C (PC), lupus anticoagulant (LA), anticardiolipin antibody) of a subset of 83 patients hyperhomocysteinemia was the most prevalent risk (33.7%), with PC or PS deficiencies following (22.9%). Thus, hyperhomocysteinemia is a prominent risk for DVT in Taiwan. PMID- 11395125 TI - Similar PAI-1 expression in visceral and subcutaneous fat of postmenopausal women. AB - Recent studies showed that intraabdominal visceral fat located in the mesenterium and omentum may significantly influence the circulating plasminogen activator inhibitor type 1 (PAI-1). To substantiate this link, we performed analysis of PAI 1 expression in human visceral and subcutaneous adipose tissues in peri- and postmenopausal women. The samples of both visceral and subcutaneous fat from 28 generally healthy women (aged 45-69 years) with a wide range of body mass index (BMI; 22.30-38.67 kg/m2), who underwent surgical operation due to benign ovary and uterine tumours, were obtained. In these samples, expression of mRNAs for PAI 1, tumour necrosis factor alpha (TNFalpha), acyl-CoA synthetase (ACS), and glucose transporter (GLUT-4) was analysed by relative quantitative RT PCR and correlated with plasma PAI-1 antigen. In addition, visceral fat area was measured with computer tomography. Both types of fat tissues contained similar quantities of PAI-1 mRNA, and there was no correlation between plasma PAI-1, measured both by antigen and activity, with either visceral or subcutaneous fat PAI-1 mRNA. Furthermore, there was no significant association between the expression of PAI-1 mRNA and TNFalpha mRNA in tested fat samples. PAI-1 mRNA in both fat tissues was significantly correlated with plasma levels of estradiol (positive correlation) and follicle-stimulating hormone (FSH; negative correlation). Finally, the expression of PAI-1 mRNA was negatively correlated with mRNA of ACS present in both fat tissues. In summary, this study directly indicates that PAI-1 mRNA is similarly expressed in both subcutaneous and visceral fat of peri- and postmenopausal women and its expression strongly depends upon lipid metabolism. PMID- 11395126 TI - Acquired von Willebrand disease in an infant. Resolution by interventional occlusion of patent ductus arteriosus. PMID- 11395127 TI - Differential effects of c7E3 Fab on thrombus formation and rt-PA-Mediated thrombolysis under flow conditions. AB - Although the Fab fragment of the mouse-human chimeric anti-alphaIIbbeta3 (GP IIb/IIIa) monoclonal antibody (MoAb) c7E3 facilitates recombinant tissue-type plasminogen activator (rt-PA)-mediated thrombolysis, it is not clear whether this is due to inhibition of new clot formation and/or a direct effect on the lysis rate. We employed an in vitro flow (re)circulation model to investigate how c7E3 Fab affected (a) platelet adhesion to clotted fibrin substrates under laminar flow at wall shear rates of 100 or 500 s(-1) and (b) rt-PA-induced lysis of preformed mural platelet-fibrin substrates at 500 s(-1). c7E3 Fab dose dependently (0.5-5 microg/ml) inhibited platelet adhesion from flowing whole blood onto fibrin substrates ( approximately 14 microm thick) at each wall shear rate. When at 5 min after the onset of flow, c7E3 Fab (0.1-10 microg/ml) and rt PA (1 microg/ml) were coinjected in flowing blood, it was found that modest fibrinolysis caused major platelet release from fibrin substrates and there was no difference in the lysis rate in the presence of rt-PA + c7E3 Fab compared to rt-PA alone. Platelet pretreatment with c7E3 Fab (10 microg/ml) had no effect on the lysis rate of thin ( approximately 40 microm), and slightly delayed the lysis rate of thick (< 250 microm), platelet-fibrin substrates containing evenly dispersed platelets (10(9)/ml). When the platelets within thick platelet-fibrin substrates were organized in platelet-rich regions ("residual thrombi"), these substrates followed a nonuniform lysis pattern, where fibrin between the thrombi lysed first and the residual thrombi lysed at a slower rate. Platelet pretreatment with c7E3 Fab (10 microg/ml) abolished the formation of the lytic resistant residual thrombi and the associated platelet-protected fibrin zones. Hence, treatment with c7E3 Fab has no direct effect on the rate of rt-PA-mediated lysis, but is expected to block platelet-fibrin interactions that lead to clot retraction, thus maintaining a fibrin architecture that is more susceptible to lysis. PMID- 11395128 TI - A prothrombin activator serine protease from the Lonomia obliqua caterpillar venom (Lopap) biochemical characterization. AB - Lonomia obliqua venom causes a severe consumptive coagulopathy, which can lead to a hemorrhagic syndrome. The crude bristles extract displays a procoagulant activity due to a Factor X and to a prothrombin activating activity. Here, we describe a 69 kDa prothrombin activator serine protease purified from L. obliqua caterpillar bristle extract using gel filtration (Sephadex G 75) and HPLC (C(4) column). The purified protein was able to activate prothrombin in a dose dependent manner, and calcium ions increased this activity. The prothrombin derived fluorogenic peptide (Abz-YQTFFNPRTGSQ-EDDnp) had its main cleavage site at the Arg-Thr bond. The kinetic parameters obtained for this substrate were Kmapp of 4.5 microM, kcat of 5.32 s(-1), and a kcat/Kmapp of 1.2 x 10(6) M(-1) s( 1). The prothrombin fragments generated by the purified enzyme corresponded to the molecular masses of prethrombin 2, fragment 1, fragment 2, and thrombin as seen in SDS-PAGE. The thrombin generated was able to clot purified fibrinogen. The partial amino acid sequence of the purified protein, named Lopap (L. obliqua prothrombin activator protease), showed no similarity to any known prothrombin activator. PMID- 11395129 TI - In vivo characterization of Lopap, a prothrombin activator serine protease from the Lonomia obliqua caterpillar venom. AB - Increasing occurrence of hemorrhagic syndrome in man, caused by contact with Lonomia obliqua caterpillars, has been reported in Southern Brazil in the past 10 years. The L. obliqua venom causes a severe consumptive coagulopathy, which can lead to hemorrhagic syndrome. L. obliqua prothrombin activator protease (Lopap) is a 69-kDa prothrombin activator serine protease isolated from L. obliqua caterpillar bristle extract, which is able to evoke thrombus formation, unclottable blood, and fibrinogen depletion when injected into the blood stream of rats. The purified protein generated thrombin from prothrombin, able to clot purified human fibrinogen and plasma. A decrease in platelet count and inhibition of collagen-induced platelet aggregation were observed, as well as leukocyte infiltration in the lungs. In addition, we observed congestion and hemorrhage in renal glomeruli and necrosis in renal distal tubules. These data support the hypothesis that Lopap contributes to the clinical syndrome caused by human contact with L. obliqua, most probably through prothrombin activation, resulting in a consumption coagulopathy. PMID- 11395130 TI - Comparison of antithrombin activity of the polysulphate chitosan derivatives in in vivo and in vitro system. AB - In order to choose the proper method for evaluating the antithrombin activity in samples of chitosan polysulphate (CP) with different polymerization degrees and sulphation degrees, we estimated the ability of direct anticoagulants to depress the coagulability of recalcified sheep blood using the third international heparin standard (A1 - in vitro system) and determined such activity on pharmacodynamic curve (A2 - in vivo system). The curve admits the kinetics of CP elimination to be nonlinear in case of intravenous injection to rabbits, as it is observed in heparin: Ct = C(o)exp(-K(e)lt), where Ct is the CP concentration at the time moment t; C(o) is the CP concentration at the injection moment; Kel is the elimination constant. Besides, it is assumed that there is a linear approximation of the anticoagulant effect on the dose, which finally makes it possible to calculate the specific activity A2: T = KTCt+T(in), where T is the time of clot formation at different time intervals after CP injection; T(in) is the time of clot formation prior to CP injection. T value was assessed in two tests: blood coagulation time (BCT) and activated partial thromboplastin time (APTT). No correlation was observed between A1 and A2. At the same time, the values of Kel and the period of semi-elimination, with the use of the biospecific cetylpyridinium chloride electrophores for the quantitative determination of CP in rabbit's blood taken at different time intervals after injection, showed a close correlation (r = .94, P < .05) between the same parameters, obtained with the help of the rectilinear pharmacodynamic plot in BCT test. Thus, experimentally, it was proven that the assumption of the CP nonlinear elimination and the CP effect-dose dependence was true, which is necessary for A2 calculation. Relatively low molecular weights (MW 61-82 kDa, polymerization degree 188-252 ) and high sulphation patterns (sulphur amounts 15.6-16.9%, sulphation degree 1.58-1.86) were slowly cleared and there was more antithrombin activity (30-52 IU/mg). We recommend the use of in vivo system for evaluating the antithrombin activity of the CP derivatives. PMID- 11395131 TI - Antithrombin activity of an algal polysaccharide. AB - In an effort to reduce the risks of a possible iatrogenic transmission of bovine spongiform encephalitis (BSE) through the use of bovine-derived medicinal products, we patented in the USA in 1999 a polysaccharide from brown algae, endowed with interesting pharmacological activities: (a) concentration-dependent inhibition of thromboplastin or cephalin-kaolin-induced thrombin generation from platelets, (b) concentration-dependent inhibition of thrombin-induced platelet aggregation, (c) thrombin has hypotensive effect, which was blunted and zeroed by our fucansulfate in a dose-dependent way, (d) when aortae are stimulated with thrombin, they become stickier for polymorphonucleated leukocytes (PMNs); our fucansulfate decreased concentration-dependently, PMNs sticking to autologous rabbit aortae, (e) dose-dependent inhibition of thrombin-induced thrombosis. All the above data suggest that our fucansulfate could be a heparin substitute endowed with antithrombotic and anti-inflammatory activities, devoid or the problems caused to heparin by its animal origin, i.e., possible prion protein contamination. PMID- 11395132 TI - Proteolysis of von Willebrand factor is increased during cardiopulmonary bypass. PMID- 11395133 TI - Fine specificity analysis of an HLA-A2.1-restricted immunodominant T cell epitope derived from human alpha-fetoprotein. AB - Human alpha-fetoprotein (AFP) is a potentially important target for the immunotherapy of hepatocellular carcinoma (HCC). AFP(542-550) (GVALQTMKQ) is one of several HLA-A2.1-restricted immunodominant AFP peptides that consistently generate AFP-specific T cell responses in human T cell cultures and in HLA A2.1/K(b) transgenic (A2.1 tg) mice. We performed a fine specificity analysis of this nonamer to determine which amino acid side chains were critical for T cell priming and recognition. Using peptide-pulsed dendritic cells (DC) as an immunization strategy, we characterized the effects of AFP(542-550) amino acid substitutions on priming and recognition in A2.1 tg mice. Replacing the glutamine at anchor position 9 with a leucine enhanced MHC binding and AFP-specific T cell responses. Substitution of leucine at non-anchor position 4 with an alanine did not alter binding but greatly diminished T cell recognition. Computer-generated three-dimensional models provided the structural rationale for these observed effects in MHC binding and T cell responses resulted from the modifications in the AFP(542-550) sequence. PMID- 11395134 TI - Prevention of experimental autoimmune encephalomyelitis by encephalitogenic epitope sequence simplified derivatives. AB - The encephalitogenic epitope P81-100 from mouse myelin basic protein was used to generate two simplified derivatives with glycine substitutions in alternating positions which were tested for their biological activity in a murine model of multiple sclerosis, experimental autoimmune encephalomyelitis. While both derivatives were unable to induce in mice the disease at the same parent peptide P81-100 dosage, T cell proliferation assays demonstrated their ability to compete with the parental peptide in a dose related manner. Experiments of cell surface binding and T cell tolerance revealed a different behavior of the two derivatives, suggesting different roles in the MHC blockade or T cell tolerance. On induction of encephalomyelitis in animals by P81-100 treatment, one variant proved in vivo to be very effective in protecting from the disease. PMID- 11395135 TI - Identification and characterization of two CD40-inducible enhancers in the mouse TRAF1 gene locus. AB - We have shown that CD40 engagement induces TRAF1 gene expression in B lymphocytes. Here we report that CD40-dependent TRAF1 gene transcription in murine B cells is controlled by two enhancer regions. One region is located approximately 2 kb upstream of the transcription start site and the other lies in the intron between exons 5 and 6. The upstream enhancer contains a single NF kappaB site in addition to sites that bind constitutive transcription factors. Mutation of this NF-kappaB site completely abrogates CD40-driven TRAFl transcription. The intronic enhancer contains two sites that strongly bind the CD40-inducible factors NF-kappaB and AP-1. Simultaneous mutation of the AP-1 site and of the NF-kappaB site abolishes transcription driven by this enhancer. When cloned together into reporter constructs, the two TRAF1 enhancers do not synergize, suggesting that each enhancer may separately participate in the induction of TRAF1 transcription in B cells following CD40 activation. PMID- 11395136 TI - Molecular analysis of peptides from the GH loop of foot-and-mouth disease virus C S30 using surface plasmon resonance: a role for kinetic rate constants. AB - A foot-and-mouth disease virus (FMDV) field variant, isolate C-S30 (also named C(1)-Barcelona), is known to contain four changes within the main antigenic site A (GH loop of capsid protein VP1, residues 136-150), at least one of which (Leu147-->Val) involves a highly conserved position, critical for antibody recognition in the reference strain C-S8c1. However, immunoenzymatic analysis of FMDV C-S30 showed it was recognised by 4C4, a monoclonal antibody that specifically targets site A. This remarkable behaviour has led us to analyse the individual and combined contributions of the four mutations to the antigenicity of C-S30, by surface plasmon resonance (SPR) and enzyme-linked immunosorbent assay (ELISA) studies of pentadecapeptides displaying all possible combinations of the four replacements. Analysis of this family of C-S30-derived analogues shows a certain level of antibody recognition by SPR. In addition, SPR data suggest that kinetic rate constants provide an indirect measure, on the one hand, of paratope accessibility (association rate constant) and, on the other hand, of peptide fitness to the same paratope (dissociation rate constant). PMID- 11395137 TI - Detection of viral superantigen-class II MHC interactions at the cell surface. AB - Mouse mammary tumor virus superantigens (vSAgs) must bind to class II MHC proteins to activate T cells. Although direct interaction of vSAgs with class II proteins has been demonstrated biochemically, the details of this interaction are largely unresolved. To facilitate the study of class II-vSAg interactions, a sensitive assay has been developed that can detect binding of vSAgs to class II proteins on the cell surface. The assay measures changes in vSAg surface expression upon enzymatic removal of a co-expressed glycan-phosphatidyl inositol anchored form of the class II molecule IE(k). Because the vSAgs are synthesized as integral membrane proteins that undergo proteolytic processing, an event that is likely required to eliminate membrane tethering, the data provide further evidence that a proteolytic fragment of vSAg is bound to class II proteins on the cell surface. The assay was utilized to identify mutant vSAgs that either did not associate with IE(k) molecules, or did not undergo furin-dependent proteolytic processing. Class II protein binding was detected using vSAg7 mutants that lacked furin endoprotease recognition sites, and after expression of vSAg in furin deficient cells. The data demonstrate that furin-mediated processing is not necessary for association of vSAg7 with class II proteins, supporting previous studies that have indicated a role for alternative endoproteases in vSAg activation. However, because class II interactions were also noted in the apparent absence of proteases that are known to activate vSAgs, the data suggest that yet other proteases may process vSAgs in a fashion that does not necessarily lead to activation of T cells. PMID- 11395138 TI - Lysine 322 in the human IgG3 C(H)2 domain is crucial for antibody dependent complement activation. AB - The classical complement activation cascade of the immune system is initiated by multivalent binding of its first component, C1q, to the Fc region of immunoglobulins in immune complexes. The C1q binding site on mouse IgG2b has been shown to contain the amino acids Glu 318, Lys 320 and Lys 322 in the C(H)2 domain (Duncan, A.R., Winter, G.,1988. The binding site for C1q on IgG. Nature 322 738 740). Identical or closely related motifs are found on all IgGs in all species, and the binding site has therefore been thought to be universal. However, the results from another study indicate that the site is different in human IgG1 molecules (Morgan, A., Jones, N.D., Nesbitt, A.M., et al., 1995. The N-terminal end of the C(H)2 domain of chimeric human IgG1 anti-HLA-DR is necessary for C1q, Fc gamma RI and Fc gamma RIII binding. Immunology 86 319-324). To determine the site(s) responsible for complement activation in anti-NIP-mouse/human IgG3 antibodies, we have mutated amino acids Lys 276, Tyr 278, Asp 280, Glu 318, Lys 320 and Lys 322 in two beta-strands in the C(H)2 domains of human IgG3. In addition, we mutated the Glu 333, which resides in close proximity to the postulated C1q-binding site of mouse IgG2b, as well as Leu 235 in the lower hinge region. All mutants were tested in Antibody Dependent Complement Mediated Lysis (ADCML)(4) assays, where the antigen concentration on target cells was varied and human serum was complement source. Only the mutants that lacked the positively charged side chain of lysine in position 322 showed strong reduction in ADCML, particularly at low antigen density on target cells. Alanine scanning of positions 318 and 320 did not affect ADCML, contrary to what was observed for mouse IgG2b. Neither did a leucine to glutamic acid mutation in position 235 have the effect that has been reported for human IgG1. These results suggest that the complement binding site on human IgG3 molecules is different from that found on mouse IgG2b, and possibly on human IgG1 as well. Thus the contact site may not be conserved. PMID- 11395139 TI - Pairing of Vbeta6 with certain Valpha2 family members prevents T cell deletion by Mtv-7 superantigen. AB - Superantigens (SAg) are proteins of bacterial or viral origin able to activate T cells by forming a trimolecular complex with both MHC class II molecules and the T cell receptor (TCR), leading to clonal deletion of reactive T cells in the thymus. SAg interact with the TCR through the beta chain variable region (Vbeta), but the TCR alpha chain has been shown to have an influence on the T cell reactivity. We have investigated here the role of the TCR alpha chain in the modulation of T cell reactivity to Mtv-7 SAg by comparing the peripheral usage of Valpha2 in Vbeta6(+) (SAg-reactive) and Vbeta8.2(+) (SAg non-reactive) T cells, in either BALB/D2 (Mtv-7(+)) or BALB/c (Mtv-7(-)) mice. The results show, first, that pairing of Vbeta6 with certain Valpha2 family members prevents T cell deletion by Mtv-7 SAg. Second, there is a strikingly different distribution of the Valpha2 family members in CD4 and CD8 populations of Vbeta6 but not of Vbeta8.2 T cells, irrespective of the presence of Mtv-7 SAg. Third, the alpha chain may play a role in the overall stability of the TCR/SAg/MHC complex. Taken together, these results suggest that the Valpha domain contributes to the selective process by its role in the TCR reactivity to SAg/MHC class II complexes, most likely by influencing the orientation of the Vbeta domain in the TCR alphabeta heterodimer. PMID- 11395141 TI - Measurement of allergenic potency using the local lymph node assay. AB - Chemicals that can act as contact allergens have been identified successfully using guinea-pig models. However, contact allergy is still common, probably because of, at least in part, failures of risk assessment. A new method, the local lymph node assay, replaces the guinea-pig as a tool for hazard identification and offers the real prospect of accurate prediction of allergen potency, the missing link in skin sensitization risk assessment. PMID- 11395153 TI - Is paradoxical pharmacology a strategy worth pursuing? AB - The first rule of medicine is, 'do no harm'. Perhaps accepting this precept has produced a logic for disease treatment where our primary purpose is limited to only assisting or helping to correct a malfunctioning system. Can we use drugs that, according to traditional views, would be considered to increase stress on the system in the short term, to actually treat and cure disease in the long term? Is it possible to exacerbate disease for a longer-term gain? Although there are several examples of where this strategy has appeared to work, a systematic testing of the hypothesis has not occurred and, for the majority of diseases, this hypothesis has never been tested. PMID- 11395154 TI - The role of toxicology in evaluating priorities in environmental health. AB - It is easy to assume that the straightforward explanation of a rational standpoint, based on sound scientific information, will inform public debate in areas of hazard evaluation. However, although toxicology generally provides the basic data that give reassurance, it is also used as a methodology for hazard identification. Once a hazard has been identified, pressure groups will present data that suit their case and ignore those that do not. Any pretence at analysis is abandoned, as shown by the recent MMR (measles--mumps--rubella) vaccine debate in the UK. Toxicologists should not suppose that comparative evaluations of risk, based on data, will result in a rational choice about environmental interventions -- data are few and opinions are a matter of faith. Thus, we might have to be protagonists in a propaganda war for science if irrational misuse of resources is to be avoided. PMID- 11395155 TI - Key challenges for toxicologists in the 21st century. AB - The application of genomic technology to toxicology (toxicogenomics) has allowed the simultaneous identification of modified gene expression in response to a toxicant to be established for thousands of mammalian genes. This, together with the development of proteomics, metabonomics and our increasing understanding of individual human polymorphisms, will enable toxicologists in the next century to identify those individuals at particular risk from specific toxins, pesticides and pharmaceutical drugs. However, these new opportunities will prove fruitless unless toxicologists address some of the major issues that presently confront their discipline. If anything, the new technologies impose a greater demand on toxicologists to exercise expert judgement on the meaning of their data, and to apply 'common sense' when balancing risks and benefits. PMID- 11395156 TI - Hormesis: U-shaped dose responses and their centrality in toxicology. AB - The fundamental nature of the dose response is neither linear or threshold, but rather U-shaped. When studies are properly designed to evaluate biological activity below the traditional toxicological threshold, low-dose stimulatory responses are observed with high frequency and display specific quantitative features. With a few exceptions, the low-dose stimulatory response is usually not more than twofold greater than the control response, with a stimulatory zone that is more variable, ranging from less than tenfold to more than several orders of magnitude of the dose. Considerable mechanistic evidence indicates that hormetic effects represent overcompensation in response to disruptions in homeostasis that are mediated by agonist concentration gradients with different affinities for stimulatory and inhibitory regulatory pathways. PMID- 11395157 TI - St John's wort: Prozac from the plant kingdom. AB - Conventional antidepressants are associated with a range of adverse drug reactions. The herb Hypericum perforatum (St John's wort) might offer another approach to the treatment of depression. Biochemical and animal studies suggest that the phloroglucinol derivative hyperforin is the main active ingredient of St John's wort, and inhibits the synaptosomal uptake of 5-HT, noradrenaline, dopamine, glutamate and GABA. St John's wort has been shown to alleviate symptoms of mild to moderate depression, and seems to offer significant advantages over conventional antidepressants because it is associated with fewer adverse reactions. However, important herb--drug interactions have been described. In view of its efficacy and safety records, St John's wort should be considered for the first-line treatment of mild to moderate depression. PMID- 11395158 TI - Genetic susceptibility to adverse drug reactions. AB - Adverse drug reactions (ADRs) are a major clinical problem. Genetic factors can determine individual susceptibility to both dose-dependent and dose-independent ADRs. Determinants of susceptibility include kinetic factors, such as gene polymorphisms in cytochrome P450 enzymes, and dynamic factors, such as polymorphisms in drug targets. The relative importance of these factors will depend on the nature of the ADR; however, it is likely that more than one gene will be involved in most instances. In the future, whole genome single nucleotide polymorphism (SNP) profiling might allow an unbiased method of determining genetic predisposing factors for ADRs, but might be limited by the lack of adequate numbers of patient samples. The overall clinical utility of genotyping in preventing ADRs needs to be proven by the use of prospective randomized controlled clinical trials. PMID- 11395159 TI - Apoptotic death sensor: an organelle's alter ego? AB - Caspases are intracellular cysteine proteases that are primarily responsible for the stereotypic morphological and biochemical changes that are associated with apoptosis. Caspases are often activated by the apoptotic protease-activating factor 1 (APAF-1) apoptosome, a complex that is formed following mitochondrial release of cytochrome c in response to many death-inducing stimuli. Both pro- and anti-apoptotic BCL-2 family members regulate apoptosis, primarily by their effects on mitochondria, whereas many inhibitor of apoptosis proteins (IAPs) regulate apoptosis by directly inhibiting distinct caspases. Exposure of cells to chemicals and radiation, as well as loss of trophic stimuli, perturb cellular homeostasis and, depending on the type of cellular stress, particular or multiple organelles appear to 'sense' the damage and signal the cell to undergo apoptosis by stimulating the formation of unique and/or common caspase-activating complexes. PMID- 11395160 TI - Toxicology of simple and complex mixtures. AB - Humans are exposed to mixtures of chemicals, rather than to individual chemicals. From a public health point of view, it is most relevant to answer the question of whether or not the components in a mixture interact in a way that results in an increase in their overall effect compared with the sum of the effects of the individual components. In this article, options for the hazard identification and risk assessment of simple and complex chemical mixtures will be discussed. In addition, key research needed to continue the development of hazard characterization of chemical mixtures will be described. Clearly, more collaboration among toxicologists, model developers and pharmacologists will be necessary. PMID- 11395161 TI - The science of pharmacology: a plea for joining forces. PMID- 11395164 TI - Editorial. PMID- 11395162 TI - Toxicologists -- come out and educate! AB - Toxicologists must be aware of the poor level of understanding among the non scientific press and media, and presumably also within governments, of the quantitative aspects of environmental (including dietary) chemical hazards. The ignorance of the basic principles of dose response, and of the differences between hazard and risk and between a chance association and a causal relationship results in huge amounts of public money being misspent by focusing on the reduction or elimination of many trivial or even imaginary hazards. Toxicologists as a profession should give more effort to the promulgation of these concepts to journalists, the general public and, particularly, those responsible for risk management. PMID- 11395165 TI - Laudatio to Professor Dr Dieter Platt on the occasion of his 65th birthday/May 2001. PMID- 11395166 TI - Metabolism and aging in the filamentous fungus Podospora anserina. AB - In Podospora anserina, lifespan is under the control of environmental and genetic factors. Both suggest an important impact of metabolism on lifespan and aging. Environmental changes of temperature, of the carbon source in the growth medium, or the addition of specific inhibitors to the growth medium are some of the investigated factors. Genetic approaches underscore the significance of metabolism. In particular, the mitochondrial electron transport plays a major role. As a by-product of a cytochrome oxidase (COX) dependent energy transduction, reactive oxygen species (ROS) are generated and lead to damage of cellular biomolecules. Damaged mitochondria, compromised at complex IV (COX) of the respiratory chain, signal to the nucleus and induce a nuclear gene, PaAox, encoding an alternative oxidase (AOX). This pathway resembles the retrograde response that, at least in yeast, is induced by dysfunctional mitochondria. ROS generation is lowered when electrons are transferred via an alternative pathway utilizing the AOX. As a consequence, lifespan of the corresponding strains is increased. Cellular copper levels were found to play a significant role not only in the generation of ROS but also have an impact on the cytoplasmic and the mitochondrial superoxide dismutase (SOD). In addition, copper is involved in the control of mitochondrial DNA rearrangements and affects the ability of the system to remodel damaged mitochondria. All these different components and pathways are part of the complex molecular network involved in lifespan control of this aging model. PMID- 11395167 TI - Dynamics of gene expression for immediate early- and late genes after seizure activity in aged rats. AB - The ability of the rodent brain to support plasticity-related phenomena declines with increasing age. A decreased coordination of genes implicated in brain plasticity may be one factor contributing to this decline. Synaptic rearrangement that occurs after seizure activity is regarded as a model of brain plasticity. In a rat model of seizure-related brain plasticity, we found that the induction of immediate-early genes, as exemplified by c-fos and tissue plasminogen activator ( tPA), is not impaired in the aged rat brain. However, the aged rat brain responded more slowly to chemically induced seizure, and the levels of c-fos and tPA mRNAs induction are decreased in the cortex and in the hippocampus of 30 month old rats, as compared to the levels expressed by 3 month old rats. In addition, at the peak induction, the TPA transcripts were restricted to certain cortical layers of the older rats. Surprisingly, in applying the same experimental paradigm to late genes, we found that there was a shift toward earlier times in the maximum expression of growth-related molecules, the microtubule-associated protein 1B (MAP1B) mRNA, which was very evident in 18 month old rats. Aberrant immunolabeling of MAP1B occurred in cortical layer VI of the aged rats where, unlike in young rats, there was heavy staining of neuronal somata. These results suggest that (1) one consequence of aging, besides decreases in the levels of mRNA, is a progressive loss of coordination in gene activity following the administration of a stimulus; (2) since c-fos, TPA and MAP1B have been implicated in neuronal plasticity, these findings could explain, in part, the limited plasticity of the aging brain. PMID- 11395168 TI - Energetic stress induces premature aging of diploid human fibroblasts (Wi-38) in vitro. AB - Oxidative phosphorylation is the main endogenous source for the generation of reactive oxygen species (ROS). In order to investigate the influence of enhanced ROS production on the in vitro senescence of Wi-38 fibroblasts, cells were cultivated in medium with elevated (hypertonic) NaCl concentrations. The number of active Na(+)/K(+)-ATPase molecules per cell was found to be increased. A rise in both respiration and glycolysis as evidenced by the increases in oxygen and glucose consumption and lactate production was revealed. Cells stayed alive in medium with NaCl concentrations of up to 0.30 M and could be adapted to growth under these hypertonic conditions (high-NaCl tolerant cells). These cells exhibited an increased cell size and protein content. A growing number of cells showed stress fibers and granulation. The proliferation rate and the maximum number of cumulative population doublings of these high-NaCl tolerant cultures were reduced and saturation density was decreased. Thus, these cells under energetic stress due to increased energy requirements for active ion transport expressed features typical for aging in vitro. We conclude therefore that energetic stress induces premature aging in human diploid fibroblasts. PMID- 11395169 TI - Age-related changes in cells and tissues due to advanced glycation end products (AGEs). AB - Advanced glycation end products (AGEs) formed by nonenzymatic glycation and oxidation of proteins accumulate during normal aging and at accelerated rate during the course of diabetes. They play a role in the pathogenesis of several other chronic diseases such as Alzheimer's disease, arthritis, atherosclerosis, pulmonary fibrosis and renal failure. AGE-formation changes the chemical and biological properties of proteins inside and outside of the cell. Binding to specific cell surface receptors induces activation of cellular signaling pathways leading to cellular dysfunction and cell death. AGEs are inducible by oxidative stress and induce oxidative stress. Subject of current studies of cell biologists is the intracellular processing of AGEs, which is accompanied by changes of the endolysosomal compartment. PMID- 11395170 TI - The effect of L-carnitine on T-maze learning ability in aged rats. AB - L-carnitine is of considerable interest because of its capacity to counteract several physiological and pathological phenomena typical of brain aging processes. We examined the effects of L-carnitine on the learning ability of old rats. 100 mg/kg per body weight per day L-carnitine was administered orally to old (21 months) male Sprague-Dawley rats (OLD-CAR) for a period of 2 months. Old (21 months, OLD-CO) and young (7 months, YG-CO) control animals received tap water exclusively. Performance of the OLD-CAR and OLD-CO was compared with that of YG-CO in a multiple T-maze. The mean run time values showed a significant (P=0.01) difference of the OLD-CAR rats to the OLD-CO but no significant differences between OLD-CAR and YG-CO. For the T-maze parameter mean correct responses we were able to demonstrate that L-carnitine treated old rats made significantly (P=0.03) less errors and significantly (P=0.01) more animals reached the T-maze goal compared with OLD-CO but no significant differences were observed between OLD-CAR and YG-CO. The results of the present study clearly demonstrate that carnitine treatment improves the learning ability of old rats and seems to be able to reduce the loss of cognitive functions that occur with aging. PMID- 11395171 TI - The aging liver: consequences for drug treatment in old age. AB - Numerous age-related changes in hepatic structure and function have been described although liver function seems to be quite well maintained in old age. Few consistent and reproducible observations and a lack of correlation between structural and functional data characterize the present state of our knowledge. However, a decline in liver volume and blood flow has been shown in older subjects, which is the physiologic basis of reduced hepatic drug clearance in this age group. In contrast to renal clearance, no reliable method exists to estimate hepatic drug clearance. The contribution of age to altered drug clearance in the elderly is difficult to assess as drug interactions, numbers and types of drugs taken at a time, underlying disease and increased interindividual variability are superimposed to the aging process. Therefore, after decades of research into the matter, the old and well known aphorism 'start lower - go slower' is valid more than ever in the field of geriatric prescribing. Not only renally excreted drugs but also substances metabolized and excreted by the liver should be used in a starting dose which is 30-40% smaller than the average dose used in middle aged adults. PMID- 11395172 TI - The influence of age on the pharmacokinetics and pharmacodynamics of bemetizide and triamterene: a single and multiple dose study. AB - Diuretics are a frequent cause of adverse drug effects in the elderly, many times involving drug-drug interactions. In addition, multiple chronic diseases, age dependent pharmacokinetic and pharmacodynamic changes, and a decreased homeostatic capacity often complicate diuretic therapy in the elderly. The pharmacokinetics (area under the plasma concentration-time curve: AUC; peak concentration in plasma: c(max); time to reach peak concentration: t(max); terminal half-life: t(1/2)) and pharmacodynamics (urine flow rates and renal excretion rates of Na(+) at 1, 3, and 6 h after oral administration) of a fixed combination of 25 mg bemetizide and 50 mg triamterene were investigated in 15 elderly patients (age 70-84 years) and 10 young volunteers (age 18-30 years) after a single dose (day 1) and after multiple doses (at steady state, day 8). Compared with the young volunteers, mean plasma concentrations of bemetizide, triamterene, and the active triamterene metabolite were significantly higher in the elderly volunteers. These elevated plasma levels occurred after single dose and were even more pronounced after multiple dose in the elderly. While plasma concentrations and AUC of bemetizide, triamterene, and the active metabolite of triamterene were increasing in correlation to age of subjects and duration of therapy, urine flow and renal Na(+) excretion rates were decreasing at the same degree. At steady state conditions, practically no effect on urine flow and Na(+) excretion rates could be observed in the elderly patients (in contrast to the young volunteers) for the first 8 h after administration of bemetizide and triamterene. The lower the measured (endogenous) creatinine clearance was in all subjects, the higher were the plasma concentrations of bemetizide and triamterene, and the lower was the effect on pharmacodynamics (i.e. urine flow and renal Na(+) excretion rates). The glomerular filtration rate, known to be lower in the elderly (a priori), was apparently decreased at higher levels of bemetizide and triamterene in the elderly, which may explain why there was no diuretic and saluretic effect after multiple dose in the elderly patients. PMID- 11395173 TI - Alpha-lipoic acid as a new treatment option for Alzheimer [corrected] type dementia. AB - Oxidative stress and energy depletion are characteristic biochemical hallmarks of Alzheimer's disease (AD), thus antioxidants with positive effects on glucose metabolism such as thioctic (alpha-lipoic) acid should exert positive effects in these patients. Therefore, 600 mg alpha-lipoic acid was given daily to nine patients with AD and related dementias (receiving a standard treatment with acetylcholinesterase inhibitors) in an open study over an observation period of, on avarage, 337+/-80 days. The treatment led to a stabilization of cognitive functions in the study group, demonstrated by constant scores in two neuropsychological tests (mini-mental state examination: MMSE and AD assessment scale, cognitive subscale: ADAScog). Despite the fact that this study was small and not randomized, this is the first indication that treatment with alpha-lipoic acid might be a successful 'neuroprotective' therapy option for AD and related dementias. PMID- 11395174 TI - Plasmalogen levels in serum from patients with impaired carbohydrate or lipid metabolism and in elderly subjects with normal metabolic values. AB - The precise role played by plasmalogen phospholipids (PL) of lipoproteins and cell membranes is not well understood. However, they might act as endogenous antioxidants in defending cell membranes and lipoproteins from reactive oxygen species. A decline of plasmalogen concentrations has been observed in some tissues in normal aging and in some pathologic conditions. For healthy adults, we had reported negative correlations of age with serum plasmalogen PL derived heaxadecanal dimethylacetal (16:0DMA) or octadecanal dimethylacetal (18:0DMA) values. To mark off these age associated changes from disturbances in glucose or lipid metabolism, this study compares the 16:0DMA and 18:0DMA data of serum PL from 118 elderly subjects, aged 57-94 years, and grouped according to the disturbance of glucose or lipid metabolism. Using a new synthetic test mixture of 16:0DMA with oleic acid butylester as a quality control in gas chromatography, the highest 16:0DMA values were found in hypercholesterolemic subjects. However, related to the bulk of serum PL, were the plasmalogens possibly act as antioxidants, the highest values of 16:0DMA/PL were found in controls. A negative correlation of serum triglycerides (TG) with 16:0DMA was detected (n=118). The data suggest a closer association of low DMA values with elevated TG levels as compared to elevated plasma glucose or other serum lipid levels. PMID- 11395175 TI - Feasibility and risks of heart surgery in very elderly: analysis of 200 consecutive patients of 80 years and above. AB - A continuing increase in the number of very elderly patients with symptomatic heart disease referred for cardiac surgery has been observed in the past. Since 1990 all patients of 80 years and above have been included in a prospective study. We report on the results of the first 200 consecutive patients (mean age: 82.2+/-2.1 years). Operative procedures ranged from isolated myocardial revascularization and valve replacement to very complex operations. In a majority of patients, the operations had to be performed as urgent or emergency cases because of advanced heart disease. Overall 30 day mortality was 9.5%. More than two thirds of patients needed treatment of perioperative complications. During long-term follow-up, cardiac surgery in octogenarians has proved to be very effective with excellent functional status and quality of life. Because of favorable results elderly patients should not be denied the benefits of cardiac surgery requiring utilization of significant medical resources. PMID- 11395176 TI - The public economics of tuberculosis control. AB - This paper identifies specific sources of the failure of private markets to allocate TB control resources efficiently. These market failures, as well as the concentration of the disease amongst the poor, suggest a number of roles for public intervention. Government intervention should aim to either increase, add to, or substitute for private supply, depending on the type of market failure and the institutional capacity of the public sector. PMID- 11395177 TI - The costs of radon mitigation in domestic properties. AB - The paper examines the costs of mitigating radon levels in domestic properties using data from the radon-affected county of Northamptonshire in the United Kingdom. Cost-effectiveness of mitigation is measured in terms of the total costs per lung cancers saved per year by mitigation expenditures. The results obtained match those reported in published theoretical estimates for proposed national mitigation programmes. Mitigation in domestic properties is shown to be more effective in reducing lung cancers than a mitigation programme designed for National Health Service workplaces in Northamptonshire, when all householders discovering radon levels above 300 Bequerels per cubic metre (Bq m(-3)) are assumed to implement mitigation strategies. In the United Kingdom, however, as elsewhere, only a small percentage of those finding raised radon levels proceed to mitigation. This reduces the effectiveness of mitigation programmes in domestic properties to levels matching those in the National Health Service workplaces. The paper confirms findings in studies from Europe and the United States that a significant proportion of householders with radon levels between 200 and 300 Bq m(-3) do not implement remedial work. The paper concludes with a range of policy proposals based on the results obtained. PMID- 11395178 TI - The potential demand for an AIDS vaccine in Thailand. AB - The recent ongoing phase III clinical trial of a preventive vaccine in Thailand has prompted studies on potential demand for the vaccine among public, employers and households. This study aims to demonstrate the impact of HIV/AIDS, estimate the AIDS vaccine budget required and design the vaccination strategies for different population groups. The analysis is based on available secondary data and several assumptions on levels of secondary infections among various risk groups. Among 15 groups, we identified eight groups as potential vaccinees: Direct CSW, IDU in treatment, IDU out of treatment, male STD, transport workers, CSW indirect, conscripts and prisoners. The vaccine budget, excluding other operating expenditure, was estimated based on a single dose regimen ranging from 100 Baht (3 US dollars) to 1000 Baht (29 US dollars) per dose. A total of 1.8 17.7 million US dollars is required for non-infected catch-up population and 0.2 1.9 million US dollars for the maintenance population in the subsequent year. We foresee a relative inefficient and inequitable consumption of AIDS vaccine, which requires proper policy analysis and government interventions. Before vaccine adoption, strong preventive measures must be in place. AIDS vaccine could play an additional, not a substituting, role. A thorough understanding, a wide consultation with stakeholders and public debates are crucial steps for sound policy formulation. PMID- 11395179 TI - Scope and scale insensitivities in a contingent valuation study of risk reductions. AB - A contingent valuation study asking willingness to pay (WTP) for reducing the overall death risk as well as the risk for fatal and non-fatal injuries in road traffic accidents was performed in Sweden 1998. Different sub-samples were used to test for scale (different risk reductions) and scope (different outcomes) effects, existence of which implies that a respondent is capable of differentiating a WTP-answer accordingly. The results indicated that respondents needed some reference point for their valuation. For instance, dependent samples showed, contrary to independent ones, a significant difference between WTP of dying from any cause and in a traffic accident for the same relative but different absolute risk reduction. Regarding non-fatal traffic accidents, tests were performed comparing valuation of risk reductions for injuries with different outcomes but identical baseline risks and relative risk reductions. Similar to the case above, dependent samples differentiated their WTP and were willing to pay significantly more for a severe injury than for a slight one, which was not the case for independent samples. PMID- 11395180 TI - Determinants of customer satisfaction with the health care system, with the possibility to choose a personal physician and with a family doctor in a transition country. AB - Many Eastern and Central European counties are reforming their health care systems. The aim of this study was to determine customer satisfaction with a reformed health care system, with the possibility of free choice of a family physician and patient satisfaction with the family physician in Slovenia and their major determinants. We used a postal survey of the patients who attended their family physician's offices during the study period. We obtained an 84% response rate. Some 72.9% of the respondents were satisfied with the current organisation of health care services, 95.5% of the respondents were satisfied with the possibility of choosing their own family physician and 58% of participants were very satisfied with the level of care received from their personal family practitioners. It was shown that higher patient satisfaction with the family physician was the most powerful predictor of patients' satisfaction with the health care system. The results show that health care reform in Slovenia has a positive impact on the consumers' perceptions of health care quality, measured in terms of consumer satisfaction with the health care system, the possibility to choose a family physician and the overall satisfaction with the family physician. PMID- 11395181 TI - Apoptosis and cell death channels in prostate cancer. AB - Apoptosis, a type of programmed cell death, is a decisive mechanism in cell processes such as homeostasis, development, and many diseases including cancer. In mammals, the mechanisms that trigger and control the process of apoptosis are complex, because it has been observed that many molecules might be involved, acting in distinct ways and depending on the cellular type. The process of apoptosis is characterized by specific biochemical and morphologic changes. However, important specific messengers such as Ca(2)+ act in active proliferation as well as in apoptosis. At present, there is convincing evidence that a sustained increase in intracellular Ca(2)+ can activate cytotoxic mechanisms in various cells and tissues. Several ionic channels located in the cytoplasmic membrane might participate in the entry of calcium into the cytosol during apoptosis. Among these ionic channels, the purinoreceptors P2X and the channels of capacitative entry of calcium have been described. Pro- and anti-apoptotic molecules such as bax and bcl-2, respectively, have also been shown to participate in the process. We have recently found the activation of a Ca(2)+ permeable, nonselective cation channel of 23 pS conductance in prostatic cancer (LNCaP) exclusively in cells previously induced to apoptosis. Our findings are discussed taking into account the different ion channels that might participate in programmed cell death in prostate cancer. PMID- 11395182 TI - Neuroprotective effect of melatonin on brain damage induced by acute global cerebral ischemia in cats. AB - BACKGROUND: Melatonin has been proposed as a neuroprotective agent on the basis of its ability to function as a free radical scavenger, provided that lipoperoxidation and other free radical damage induced by reactive oxygen species resulting from cerebral ischemia are relevant pathophysiologic processes of ischemic neuronal damage. METHODS: The effects of melatonin or vehicle on neurologic deficit scores (Todd scale; maximal deficit score = 100, daily during 7 days after the ischemic episode) and neuronal population of hippocampal CA1-CA4 fields (cresyl violet stain, at the eighth day after the ischemic episode) were evaluated in adult male cats subjected to a 15-min period of acute global cerebral ischemia induced by cardiorespiratory arrest, and in cats subjected to a sham procedure. Continuous intravenous (iv) administration of either melatonin 10 mg/kg/h in 10% ethanol in saline or the vehicle alone (3 mL/kg/h) for 6 h starting 30 min after the end of the period of global cerebral ischemia was used as treatment. RESULTS: Global cerebral ischemia resulted in a severe loss of neurons in hippocampal CA1-CA4 fields (9, 13, 30 and 28% remaining neurons, respectively) of ischemic vehicle-treated cats in comparison with sham cats (100%). By contrast, remaining neurons in these regions were between 81 and 100% in the ischemic melatonin-treated cats, values that are nonsignificantly different as compared with sham cats. Values of remaining neurons in CA1-CA4 fields in ischemic melatonin-treated cats were significantly higher than those in ischemic vehicle-treated cats. Neurologic deficit scores in ischemic vehicle treated cats (42-77 at day 1, 6-39 at day 7) were significantly higher than those in ischemic melatonin-treated cats (16-38 at day 1, 0-6 at day 7) on the days after the ischemic episode. CONCLUSIONS: Overall, the results support the neuroprotective effect of melatonin against the neuronal cerebral damage induced by acute global cerebral ischemia. PMID- 11395183 TI - Additive effect of clonidine and fluoxetine on apomorphine-induced aggressive behavior in adult male Wistar rats. AB - BACKGROUND: Fluoxetine, a selective serotonin reuptake inhibitor, has been reported to be superior in its antiaggressive properties as compared with other antidepressants. The objective of the study was to investigate whether the effect of a minimal effective dose of fluoxetine could be potentiated by the alpha2 adrenergic agonist clonidine. METHODS: Vehicle (1.0 mL/kg), clonidine (0.1 mg/kg), fluoxetine (10 mg/kg), and their combination [clonidine (0.1 mg/kg) plus fluoxetine (10 mg/kg)] were injected into apomorphine-pretreated (1.0 mg/kg, once daily during 12 days) aggressive adult male Wistar rats. RESULTS: Repeated apomorphine treatment induced a gradual development of aggressive behavior. Combined clonidine and fluoxetine treatment attenuated the intensity of aggressive behavior, while these drugs alone had only a weak tendency toward reduction of aggression. Latency before the first attack was unchanged. CONCLUSIONS: Our experiments confirm that combined clonidine and fluoxetine treatment elicits an additive antiaggressive effect on apomorphine-induced aggressive behavior in rats. PMID- 11395184 TI - Peritoneal protein loss in patients with high peritoneal permeability: comparison between continuous ambulatory peritoneal dialysis and daytime intermittent peritoneal dialysis. AB - BACKGROUND: Dialysate protein loss is involved in the etiology of hypoalbuminemia and malnutrition on continuous ambulatory peritoneal dialysis (CAPD). Patients with high peritoneal membrane permeability had the lowest serum albumin (Alb) and highest dialysate protein concentrations and achieved higher small solute dialysis/plasma equilibration in a shorter time than patients with low peritoneal transport. The aim of this prospective crossover study was to evaluate whether protein loss might be decreased in patients with high peritoneal permeability on short dwell-time (DT) peritoneal dialysis. METHODS: Five high and nine high average peritoneal transport patients were subjected to the following sequential dialysis schemes (four exchanges/day, glucose 1.5%): scheme A, three daytime exchanges (4-6 h DT) and one nightly (8-12 h DT) for 2-3 days, scheme B, 3-h DT each and dry peritoneum at night during 5 days, a wash-out period similar to scheme A, and scheme C, 2-h DT each and dry peritoneum the remainder of day and night during 5 days. Dialysate Alb, IgG, IgA, and IgM losses and adequacy of dialysis were evaluated at the end of each scheme. RESULTS: Dialysate IgM was not detected. All protein losses were reduced with the short DT dialysis schemes; however, dialysis CCl and KT/V(urea) were also decreased. In patients with high peritoneal transport type, the 3-h DT dialysis scheme achieved a reduction in Alb loss without significant reduction of adequacy of dialysis. CONCLUSIONS: Peritoneal Alb, IgG, and IgA losses are significantly reduced in patients with high peritoneal permeability on short dwell-time dialysis and extended dry periods. However, a reduction of dialysis contribution to small solute clearances was also observed, Three-hour dwell-time dialysis may be particularly useful in patients with high peritoneal transport type, as it tends to reduce peritoneal protein loss without notably reducing adequacy of dialysis. PMID- 11395185 TI - Cytogenetics in acute lymphoblastic leukemia in Mexican children: an institutional experience. AB - BACKGROUND: Cytogenetic studies in acute lymphoblastic leukemia (ALL) have identified numerical and structural chromosomal abnormalities related to the disease's pathophysiologic characteristics. These findings correlate with prognosis and response to treatment in ALL patients. The purpose of this study was to define the frequency of chromosomal abnormalities in a group of Mexican children with ALL and to compare these data with those reported in the literature. METHODS: Bone marrow chromosome studies with GTG bands were performed in 150 pediatric patients with ALL who were naive to antileukemic treatment and aged from 5 months to 16 years; the majority was diagnosed as L1. RESULTS: Among 131 patients, 30 (22.9%) karyotypes were normal and the remaining 101 (77.1%) had abnormal karyotypes with numerical and/or structural abnormalities. Among patients with numerical abnormalities, the most frequent karyotypes were hyperdiploidy with 51-65 chromosomes (30 patients) and hyperdiploidy with 47-50 chromosomes (18 patients). Among recurrent, non-random, and primary structural abnormalities, the most frequent was t(9;22), followed by t(1;19). Aberrations involving band 11q23 were not detected, and only one of two patients with L3 had the t(8;14). Of the secondary non-random abnormalities, dup(1q), del(6q), and i(7)(q10) were found. CONCLUSIONS: The frequency and type of chromosomal abnormalities found was comparable to those reported in the literature with similar methodology and pediatric populations; however, the number of cases analyzed should be increased to create a database of Mexican children with ALL, and several patients require molecular analysis to identify chromosomal abnormalities not detected through conventional cytogenetic studies. PMID- 11395186 TI - Weight, physical activity, and smoking as determinants of insulinemia in adolescents. AB - BACKGROUND: It is known that insulinemia is the result of the interaction among several factors, such as weight, body fat pattern distribution, and physical activity as well as ethnicity. There is little information regarding this question among Mexican adolescents. The association among fasting insulin levels, weight, fat distribution, physical activity, and cigarette smoking was studied in Mexican adolescents. METHODS: Cross-sectional data were collected from 352 Mexican adolescents aged 14-19 years (response rate 41.5%). Fasting insulin levels were measured by microparticle enzyme immunoassay; body mass index (BMI), waist-to-hip ratio (WHR), and waist circumference (WC) were determined using standardized techniques, while physical activity was determined by 7-day Stanford physical activity inventory. Cigarette smoking was defined as number of cigarettes/year. RESULTS: Increased BMI and waist circumference, low physical activity, younger age, and non-smoking were associated with high insulin levels. Non-smokers had higher fasting insulin levels compared to smokers (57.8 pmol +/- 1.84 vs. 49.7 pmol/L +/- 2.8; p = 0.034). However, adjusted odds ratio (OR) between insulin and smoking status was not significant. Multivariate analysis showed the following: insulin increased 1.06 pmol/L for each unit of change in BMI; increased 1.02 pmol/L for each unit of change in waist circumference; increased 1.16 pmol/L for non-smoking, and decreased 1.07 pmol/L for each 250 kcal/day of energy expenditure. CONCLUSIONS: Our results support the concept that Mexican adolescents who are overweight have abdominal fat distribution and physical inactivity, and significant hyperinsulinemia. The relationship between smoking and lower insulinemia found in this research warrants further study. PMID- 11395187 TI - Event-related brain potentials during a continuous performance test (CPT) task in normal children. AB - BACKGROUND: Continuous Performance Test (CPT) is a commonly used paradigm to assess attention disorders that could involve working memory processes. METHODS: Event-related potentials (ERPs) during a CPT (X-AX) test were obtained in 16 healthy male students, with ages ranging from 9 to 11 years (X = 10.3). In grouped averaged ERPs, an endogenous slow positive potential was recorded in the first task (infrequent letter detection); maximum was at 460 msec with a slight lateralization tendency toward left parietal area. In the second task (target detection with an A as warning signal), an early (maximum at 330 msec) and more acute peak was detected without evidence of any lateralization. RESULTS: Reaction times were significantly shorter for the second task. Electrophysiologic differences between both target conditions showed an early, remarkable, and statistically significant component located at the parietal area at 340 msec. CONCLUSIONS: These ERPs findings could be interpreted as correlates with working memory processes in children. PMID- 11395188 TI - Naltrexone-induced augmentation of sexual response in men. AB - BACKGROUND: To ascertain the role of endogenous opioids in sexual response, naltrexone, an opiate receptor antagonist, was administered to men, and its effect on selected self-report measures of sexual response to masturbation was recorded. METHODS: The data are based on results from 20 healthy, sexually active (alone or with a partner) men, aged 20-29 years, who ingested naltrexone (25 mg/day x 3) or placebo in a randomized, double-blind crossover design. There was at least a 14-day interval between drug and placebo treatment. Between 18 and 22 h after the most recent dose of drug or placebo, subjects viewed sexually explicit videos in privacy for 2 h. They were instructed to masturbate and have as many orgasms as desired. The following three different self-report measures of their responses were recorded: number of orgasms; intensity of sexual arousal, and orgasmic intensity. RESULTS: Under the naltrexone condition, the volunteers experienced a significantly greater mean number of orgasms (3.4 +/- 0.2 SEM) than under the placebo condition (2.6 +/- 0.3). The total number of orgasms was 67 under the naltrexone condition and 51 under the placebo condition. At the first orgasm, the measure of intensity of arousal was significantly greater in the naltrexone (3.9 +/- 0.2) than placebo (3.4 +/- 0.2) condition, and the measure of orgasmic intensity was significantly greater in the naltrexone (3.7 +/- 0.2) than in the placebo (3.0 +/- 0.3) condition. CONCLUSIONS: The present study provides evidence that endogenous opioids modulate orgasmic response and the perceived intensity of sexual arousal and orgasm in men. The findings suggest that naltrexone could be clinically useful in cases of inhibited sexual desire and erectile dysfunction. PMID- 11395189 TI - Mild perioperative hypothermia and the risk of wound infection. AB - BACKGROUND: Bacterial destruction caused by free radicals, which are synthesized by neutrophils in the presence of oxygen, depends on adequate tissue perfusion. Mild perioperative hypothermia causes vasoconstriction, reducing nutrient and oxygen supply to wounds and increasing frequency of surgical wound infection. However, the causal role of hypothermia in surgical wound infection is the subject of controversy. The present work proposes the hypothesis that mild perioperative hypothermia is associated with infection of the surgical wound. METHODS: A prospective cohort of 290 surgical patients was studied in a second level hospital; 261 (90%) of the patients concluded the follow-up. The relationship of hypothermia and of other confounding factors, such as diabetes mellitus, antibiotic treatment, and wound drains with infection outcome was evaluated. One physician, blinded to patient hypothermia, gathered the data. Surgical wound infection was defined as the surgeon's diagnosis with positive culture. RESULTS: Twenty subjects (7.6%) showed infection of surgical wound; 18 (11.5%) of 156 hypothermics and two (2%) 105 normothermics (p = 0.004). Hypothermia proved to be a significant independent risk of infection with relative risk of 6.3 (p = 0.01). CONCLUSIONS: Mild perioperative hypothermia is associated with infection of the surgical wound and its prevention is therefore justified. PMID- 11395190 TI - Y-chromosome haplotypes for six short tandem repeats (STRs) in a Mexican population. AB - BACKGROUND: Short tandem repeats (STRs) on the non-pseudoautosomal region of the Y-chromosome are DNA polymorphic markers able to solve special cases in legal medicine, for instance in paternity testing where the alleged father is not available, and in forensic situations, such as rape cases, where mixtures of male/female DNA are present. METHODS: Six STR polymorphisms from the Y-chromosome (DYS19, DYS385, DYS389/I, DYS390, DYS391, and DYS393) were PCR-typed in 120 males from the northwest region of Mexico by means of native polyacrylamide gel electrophoresis and silver staining. RESULTS: Allele frequencies were estimated for each STR. Their gene diversity ranged from 51.4% for DYS393 to 92.5% for DYS385. Mexican Y-STR allele distributions displayed similarity (p >0.05) with previously reported U.S. Hispanics for DYS19, DYS389/I, DYS390, DYS391, and DYS393. Although Mexicans showed the same modal allele for DYS385 (11/14; 24.4%) with regard to most European populations, differences in allele distributions were observed (p <0.01). The haplotype diversity and the male discriminatory capacity of this six-locus system were 99.3 and 84.1%, respectively. CONCLUSIONS: This knowledge permits the effective use of these six Y-chromosome markers in legal medicine casework in the studied population. This STR-system offers a great potential to identify males and male-lineages, and can be used confidentially in paternity testing and forensic analysis in the Mexican population. PMID- 11395191 TI - Epidemiologic Study of Pseudomonas aeruginosa in critical patients and reservoirs. AB - BACKGROUND: Pseudomonas aeruginosa is a common cause of nosocomial infections, particularly in intensive care units (ICUs). The aim of this study was to characterize P. aeruginosa clinical isolates by comparing antimicrobial susceptibility patterns with the presence of plasmids and to establish the clonal relatedness by pulsed-field gel electrophoresis (PFGE) typing. METHODS: The patients included those with isolation of P. aeruginosa hospitalized for more than 48 h in the ICU from April to May 1998. Environmental and staff cultures were obtained simultaneously. Minimal inhibitory concentrations, plasmid DNA profiles, and PFGE genomic patterns of enzyme restriction chromosomal DNA were compared. RESULTS: Sixty P. aeruginosa isolates were obtained from 197 clinical specimens, 178 environmental samples, and 47 hand cultures of personnel. Antimicrobial resistance was as follows: tobramycin 100%; ticarcillin, cefotaxime, ceftriaxone, ceftazidime, and gentamicin 80%; cefepime 60%; amikacin, ticarcillin/clavulanate, imipenem, and meropenem 40%; piperacillin and norfloxacin 20%; carbenicillin 12%, and ciprofloxacin 0%. Plasmids were detected in 11 isolates (18%). PFGE typing showed that 23 isolates belonged to a common clone (pattern A), identified from five patients, two nurses, and 10 environmental samples. Ten isolates were grouped in four clusters and 27 isolates had unrelated genomic patterns. There was no relationship among DNA genomic patterns, plasmid profiles, and susceptibility patterns. CONCLUSIONS: PFGE demonstrated the existence of a common clone in a critical care area. Reinforcement of infection control measures is needed to avoid horizontal transmission and severe infections. PMID- 11395192 TI - Acceptability of a human papillomavirus (HPV) trial vaccine among mothers of adolescents in Cuernavaca, Mexico. AB - BACKGROUND: A scenario that must be considered when testing prophylactic human papillomavirus (HPV) vaccines in teenagers is the parents' acceptability of their daughters' participation in the study. METHODS: A survey was carried out in a random sample of 880 women between the ages of 15 and 49 years in the metropolitan area of Cuernavaca, Mexico. These women were interviewed to obtain information concerning their knowledge of risk factors for cervical cancer and their perception of the usefulness of vaccines. Afterward, they were provided with information on the main risk factors for cervical cancer and the future availability of a human papillomavirus (HPV) vaccine to prevent cervical cancer. Finally, we explored, with parents, the possible acceptability of an HPV vaccine for their teenaged daughters. The degree of acceptability and its association with a series of sociodemographic and reproductive factors were assessed. RESULTS: The respondents had little knowledge regarding the etiology of cervical cancer. Only 1.9% said that the principal risk factor was infection with HPV; however, 84.2% were aware of the usefulness of vaccines and 83.6% of the women indicated that they would allow their daughters to participate in a trial to evaluate the effectiveness of an HPV vaccine that helps prevent cervical cancer. The main factor associated with the acceptance of a possible vaccine against HPV was the knowledge of the usefulness of vaccines [odds ratio (OR) = 6.5, 95% confidence interval (CI) 5.2-8.2]. Likewise, a history of two or more sexual partners (OR = 2.2, 95% CI 1.3-3.6) increased acceptability. Acceptance was not associated with the number of live births (never vs. ever OR = 0.9, 95% CI 0.3 2.1). There were 525 women with children over the age of 10 years (59.6%); prevalence of acceptability among these women was 80.1%, not statistically different from the remainder of the sample (p >0.05). CONCLUSIONS: Acceptance of a potential HPV vaccine was high in this sample of Mexican women. Initiation of HPV vaccine clinical trials and immunization campaigns that target school children and/or teenagers who are not sexually active should include educational programs aimed at mothers of these individuals. Knowledge of the benefits of a preventive vaccine as well as the etiology and risk factors of cervical cancer should be emphasized. PMID- 11395193 TI - Development of Th1-mediated CD8+ effector T cells by vaccination with epitope peptides encapsulated in pH-sensitive liposomes. AB - There have been many studies for tumor therapy mediated by cytotoxic T lymphocytes (CTL) that recognize tumor-associated antigen. It is generally accepted that CTL responses are induced when antigen is delivered into the cytosol. The pH-sensitive liposomes as vehicles are well known for their capacity to deliver the antigen into the cytosol. In this work, immunization of mice with CTL epitope peptides from Hantaan nucleocapsid protein (M6) or human papilloma virus E7 encapsulated in pH-sensitive liposomes induced effective antigen specific CTL responses. The CTL responses induced by M6 peptide encapsulated in pH-sensitive liposomes blocked the formation of tumor mass from Hantaan NP transfected B16 melanoma cells in C57BL/6 mice and delayed the growth of preinoculated melanoma cells. During the blockade of the tumor growth, the CTL response was maintained for at least approximately 6 weeks, and the mice secreted Th1 type cytokines such as IL-2 and IFN-gamma. These results suggested that the pH-sensitive liposomes might provide an effective peptide delivery system for CTL mediated tumor therapy. PMID- 11395194 TI - Hepatitis A control in a refugee camp by active immunization. AB - An outbreak of hepatitis A occurred among children of a refugee camp in Croatia. In order to disrupt the outbreak, we decided to vaccinate children from 1 to 15 years of age in the camp, in addition to intensified general preventive measures. Assuming high prevalence of hepatitis A virus antibodies within this population, we conducted anti-HAV testing of the children eligible for vaccination. Of 108 children tested, 74 (68.5%) were anti-HAV positive. We vaccinated 34 children. One month after vaccination 31 previously negative children were tested for anti HAV and 30 of them were found positive, suggesting a seroconversion rate of 96.8%. One child fell ill 5 days after vaccination, after whom no new cases of hepatitis A occurred. Thus we conclude that active immunization is a successful means of stopping an outbreak of hepatitis A. PMID- 11395195 TI - Construction of a live-attenuated bivalent vaccine virus against human parainfluenza virus (PIV) types 1 and 2 using a recombinant PIV3 backbone. AB - PIV1 and PIV2 are important agents of pediatric respiratory tract disease. We are developing live-attenuated vaccines against these viruses. We earlier constructed a PIV3/PIV1 antigenic chimeric virus, designated rPIV3-1, in which the hemagglutinin-neuraminidase (HN) and fusion (F) proteins of wild type rPIV3 were replaced by their PIV1 counterparts. In the present study, rPIV3-1 was used as a vector to express the HN protein of PIV2 to generate a single virus capable of inducing immunity to both PIV1 and PIV2. The PIV2 HN open reading frame was expressed from an extra gene cassette, under the control of PIV3 cis-acting transcription signals, inserted between the F and HN genes of rPIV3-1. The recombinant derivative, designated rPIV3-1.2HN, was readily recovered and exhibited a level of temperature sensitivity and in vitro growth similar to that of its parental virus. The rPIV3-1.2HN virus was restricted in replication in both the upper and lower respiratory tracts of hamsters compared with rPIV3-1, identifying an attenuating effect of the PIV2 HN insert in hamsters. rPIV3-1.2HN elicited serum antibodies to both PIV1 and PIV2 and induced resistance against challenge with wild type PIV1 or PIV2. Thus, rPIV3-1.2HN, a virus attenuated solely by the insertion of the PIV2 HN gene, functioned as a live attenuated bivalent vaccine candidate against both PIV1 and PIV2. PMID- 11395196 TI - MMR and autism: further evidence against a causal association. AB - The hypothesis that MMR vaccines cause autism was first raised by reports of cases in which developmental regression occurred soon after MMR vaccination. A previous study found no evidence to support this hypothesis. It has recently been suggested that MMR vaccine might cause autism, but that the induction interval need not be short. The data from the earlier study were reanalysed to test this second hypothesis. Our results do not support this hypothesis, and provide further evidence against a causal association between MMR vaccination and autism. PMID- 11395197 TI - A quantitative assessment of the effectiveness of PRRSV vaccination in pigs under experimental conditions. AB - This paper presents a quantitative approach to evaluate effectiveness of vaccination under experimental conditions. We used two consecutive experimental designs to investigate whether PRRSV transmission among vaccinated pigs was reduced compared to control pigs and to estimate the reproduction parameter R. Based upon data analysis and power calculations the series of small-scale vaccination-challenge experiments ended with multiple one-to-one experiments. This new experimental design has considerable power to detect the effect of vaccination on transmission if R is close to but still above one in vaccinated pigs. The last experiment showed that transmission was not significantly reduced and the R for vaccinated pigs was estimated to be larger than 4.9. This is remarkable because duration and level of viremia were significantly reduced by vaccination. PMID- 11395198 TI - Impact of osmolality on burning sensations during and immediately after intramuscular injection of 0.5 ml of vaccine suspensions in healthy adults. AB - A randomised placebo controlled double-blind cross-over trial was performed on twenty healthy adults to assess the effect of osmolality (300,600,850 and 1100 mOsm) on local tolerance of an intramuscular injection (0.5 ml) of five suspensions containing the same components as the excipients of a combined Diphtheria-Tetanus-acellular Pertussis-inactivated Poliomyelitis-Haemophilus influenzae type b paediatric vaccine (DtacP-IPV-Hib, PENTAVAC). The results did not show any dose-effect relationship between burning or pain sensations and the different osmolalities tested. Although mild and not clinically relevant, these sensations seemed to occur more frequently following injection of an isotonic saline solution (P<0.05). Thus, the osmolality of vaccine like suspensions does not appear to be a potential cause of local pain or burning sensation after their administration. PMID- 11395199 TI - Pre-clinical safety and efficacy of TA-CIN, a recombinant HPV16 L2E6E7 fusion protein vaccine, in homologous and heterologous prime-boost regimens. AB - Human papillomavirus (HPV) E6 and E7 oncoproteins are attractive targets for T cell-based immunotherapy of cervical intraepithelial neoplasia (CIN) and cancer. A newly designed vaccine, comprising the HPV16 L2, E6 and E7 as a single fusion protein (TA-CIN), was shown to elicit HPV16-specific CTL, T-helper cells and antibodies in a pre-clinical mouse model. These immune responses effectively prevented outgrowth of HPV16-positive tumour cells in a prophylactic setting as well as in a minimal residual disease setting. CTL immunity was optimally induced when TA-CIN was employed in heterologous prime-boost regimens in combination with TA-HPV, a clinical grade vaccinia-based vaccine. These data provide a scientific basis for the use of TA-CIN, alone or in combination with TA-HPV in future human trials. PMID- 11395200 TI - Inactivated recombinant plant virus protects dogs from a lethal challenge with canine parvovirus. AB - A vaccine based upon a recombinant plant virus (CPMV-PARVO1), displaying a peptide derived from the VP2 capsid protein of canine parvovirus (CPV), has previously been described. To date, studies with the vaccine have utilized viable plant chimaeric particles (CVPs). In this study, CPMV-PARVO1 was inactivated by UV treatment to remove the possibility of replication of the recombinant plant virus in a plant host after manufacture of the vaccine. We show that the inactivated CVP is able to protect dogs from a lethal challenge with CPV following parenteral immunization with the vaccine. Dogs immunized with the inactivated CPMV-PARVO1 in adjuvant displayed no clinical signs of disease and shedding of CPV in faeces was limited following CPV challenge. All immunized dogs elicited high titres of peptide-specific antibody, which neutralized CPV in vitro. Levels of protection, virus shedding and VP2-specific antibody were comparable to those seen in dogs immunized with the same VP2- peptide coupled to keyhole limpet hemocyanin (KLH). Since plant virus-derived vaccines have the potential for cost-effective manufacture and are not known to replicate in mammalian cells, they represent a viable alternative to current replicating vaccine vectors for development of both human and veterinary vaccines. PMID- 11395201 TI - Anti-HBs after hepatitis B immunization with plasma-derived and recombinant DNA derived vaccines: binding to mutant HBsAg. AB - The G145R mutant of the small S-protein is a major escape mutant of hepatitis B virus observed in natural infection, after immunization and HBIG therapy. In a previous study we found that plasma-derived and recombinant DNA-derived vaccine HBsAg reacted differently with monoclonal antibodies sensitive for the G145R change. In the present study we investigated the binding of polyclonal anti-HBs obtained after immunization with plasma vaccine and recombinant DNA vaccine to synthetic peptides (adw(2), adr) and rHBsAg (HepG2) (ayw(3); wild type and a 145R mutant). Anti-HBs binding to synthetic peptids (25-mers, 7aa overlap) from the "a"-loop was significantly reduced by the G145R substitution and by changing the amino acid sequence from adw(2) into adr. With mutant G145R rHBsAg the inhibitory activity of vaccine anti-HBs was decreased compared to rHBsAg wild type. In general only minor differences were observed between plasma vaccine and recombinant DNA vaccine related antibody responses. However, the individual heterogeneity in epitope specific reactivity with its possible consequences for protection (against escape mutants) is not reflected in an anti-HBs titer by standard anti-HBs assays. The presented differentiation in anti-HBs response after immunization may deliver new tools for evaluation of future vaccines. PMID- 11395202 TI - Protective immunity against influenza A virus induced by immunization with DNA plasmid containing influenza M gene. AB - DNA vaccination is characterized by its preferential induction of the cytotoxic T cell lymphocyte (CTL) response and is expected to be a useful means of protection against viral infection. We examined the protective effect of an expression plasmid (pME18S-M) containing M1 and M2 genes of influenza A/PR/8/34. We detected the CTL activity by introducing these plasmids into BALB/c mice by either the intramuscular or the intranasal route. The influenza-specific antibody response was also induced, although its neutralizing effect against influenza virus was not observed. From 70 to 80% protection was observed in the mice immunized with the pME18S-M plasmid followed by lethal infection with influenza viruses of the A/WSN/33 and A/PR/8/34 strains, whereas all mice without the plasmid vaccination failed to survive. This protective activity was significantly weakened when the CD8(+) cells of these immunized mice were eliminated by several injections of anti-CD8 antibody. The protective activity was also weakened when anti-CD4 antibody was injected in the early phase of DNA vaccination. These data suggest that the pME18S-M plasmid is useful as a DNA vaccine for overcoming highly mutational influenza viruses. PMID- 11395203 TI - Plasmid DNA-recombinant Opc protein complexes for nasal DNA immunization. AB - The nasal mucosa may provide a simple, non-invasive route to deliver DNA encoding genes that stimulate a specific immune response. Based on this, a new approach using pCMVbeta-gal plasmid DNA complexed to the Opc meningococcal outer membrane protein was assayed for. Optimal conditions of interaction were established between recombinant Opc protein and pCMVbeta-gal plasmid DNA. Complexes were fully characterized by electrophoresis analysis, DNAse resistance assay and transmission electron microscopy. DNA-protein complexes were also evaluated in in vitro transfection experiments. After the characterisation of complexes, Balb/c mice were intranasal (i.n.) and intramuscularly (i.m.) immunized. The humoral immune response against beta-galactosidase was measured by ELISA. The proliferative response in the spleen lymph nodes was also measured. Complexes administered by i.n. route induced both systemic and mucosal antibody responses. This behavior was not observed with the naked DNA. Finally, a lymphoproliferative response specific to beta-galactosidase induced by DNA-protein complexes was also detected. PMID- 11395204 TI - Safety of modified vaccinia virus Ankara (MVA) in immune-suppressed macaques. AB - Modified vaccinia virus Ankara (MVA)-based recombinant viruses have been shown to be potent vaccine candidates for several infectious and neoplastic diseases. Since a major application of these live, replication-deficient vectors would be their use in immunocompromised or potentially immunocompromised individuals, a preclinical safety study was carried out. Macaques were inoculated with high doses of MVA (10(9)) via various routes, after immune-suppression by total-body irradiation, anti-thymocyte globulin treatment, or measles virus (MV) infection. No clinical, haematological or pathological abnormalities related to MVA inoculation were observed during a 13-day follow-up period. The presence of MVA genomes was demonstrated by nested PCR during the course of the experiment in all macaques, but from none of these animals replication competent MVA could be reisolated. These data suggest that MVA can safely be used as a basis for recombinant human vaccines, and that it is also safe for use in immunocompromised individuals. PMID- 11395205 TI - Dendritic cells generated in vivo by a chimeric hematopoietic growth factor, progenipoietin-4, demonstrate potent immunological function. AB - Recently, a dual receptor agonist for human Flt3 and G-CSF receptors, progenipoietin-4 (ProGP-4), was shown to be highly effective in expanding DC in vivo. In this study, we examined the immunological activity of ProGP-4-generated dendritic cell (DC) in an HLA-A2.1 transgenic mouse system. ProGP-4 DC were found to be approximately equivalent in presenting a cytotoxic T lymphocyte (CTL) peptide to a CTL line in vitro compared with bone marrow (BM)-derived DC and >20 fold more efficient than macrophages or B cells, and >100-fold better than BM-DC, macrophages, or B cells at presenting PADRE, a universal helper T cell epitope, to a T cell clone. The heightened epitope presentation by ProGP-4 DC was paralleled in vivo inasmuch as a >6-fold increase in CTL induction was observed compared with other APC populations following ex vivo loading with peptide. The in vitro and in vivo CTL responses stimulated by ProGP-4 DC could be further augmented by either culturing with tumor necrosis factor-alpha (TNF-alpha) or co loading with PADRE. Collectively, our results indicate that peptide-loaded ProGP 4-generated DC demonstrate potent antigenicity and immunogenicity for CTL, making them an attractive component of epitope-based vaccines. PMID- 11395206 TI - Cost of immunization with a locally produced, oral cholera vaccine in Viet Nam. AB - Policy decisions regarding whether to incorporate new vaccines into routine public health practice in developing countries will depend in part on the costs of vaccine purchase and of vaccine delivery. In March, 1997, a large-scale effectiveness trial of a locally produced, orally administered bivalent vaccine against Vibrio cholerae 01 and 0139 began in Viet Nam. Empirical data obtained from the trial was used to determine the costs of the immunization campaign from the government perspective. The study population, including the children less than one year of age and pregnant women who were ineligible for immunization, was 353926. A total of 289041 persons received two doses of vaccine, and 13340 persons received one dose of vaccine. Two-dose vaccine coverage was 83.4%. The total cost of vaccine delivery during the immunization campaign was $66527. The cost of each dose of vaccine was $0.31. Therefore, the total cost of the immunization campaign was $0.44 per dose administered, and $0.91 per fully immunized person. Attempts to reduce the cost per dose of vaccine (e.g. the use of a monovalent vaccine against serogroup 01) are likely to have a large impact on the cost of future similar immunization campaigns. PMID- 11395207 TI - A bacterially expressed peptide prevents experimental infection of primates by the hepatitis E virus. AB - A 23 kDa peptide of the major structural protein of the hepatitis E virus (HEV) expressed in E. coli was found to naturally interact with one another to form homodimers and the peptide was recognized strongly in its dimeric form by HEV reactive human sera. To determine if the peptide may confer protection against HEV infection, three monkeys were immunized with the purified peptide and three were given placebo. Both groups of animals were challenged with 10(5) genome equivalent dose of the homologous strain of HEV. All control animals excreted the virus for 10-12 days beginning 5 days after the infection. The viral genome was also present in the peripheral blood monocyte (PBMC) samples from two animals, but it was not detected in the plasma samples from any of the animals. The infection in two control animals was accompanied by HEV seroconversion. Immunization was found to abrogate HEV stool excretion in two animals and reduced the viral excretion to one day in the third. None of the immunized animals showed detectable HEV in plasma or PBMC samples nor did the animals showed evidence of HEV seroconversion. These results suggested that immunization with the bacterially expressed peptide may prevent experimental infection of primates with the homologous strain of HEV. PMID- 11395208 TI - Antibody, cytokine and cytotoxic T lymphocyte responses in chimpanzees immunized with human papillomavirus virus-like particles. AB - We evaluated antibody, cytokine (IFN-gamma, IL-5, TNF-alpha), and cytotoxic T lymphocyte (CTL) responses in chimpanzees immunized with monovalent or quadrivalent (HPV-6, -11, -16, -18) L1 virus-like particle (VLP) vaccines administered i.m. on aluminum hydroxyphosphate (alum) at weeks 0, 8 and 24. Maximum serum antibody titers to type-specific, neutralizing, conformational epitopes on HPV-11 or -16 L1 VLPs were detected by radioimmunoassay (RIA) four weeks after the second and third immunizations. HPV-11 and -16 neutralizing antibodies were also detected at similar time points with an Human papillomaviruses (HPV) neutralization assay using pseudovirions. Depending on the VLP type used for immunization, HPV type-specific cytokine responses were most frequently seen four weeks after the second or third immunizations and between weeks 44-52. Transient HPV-16 L1-specific CTL activity was observed only between weeks 16-24 in 3 of 22 (13.6%) chimpanzees immunized with HPV-16 L1 VLPs. These findings provide evidence that immunization with multivalent L1 VLPs on alum can evoke both neutralizing antibodies and Th1 and Th2 cytokine responses to several HPV types; however, induction of CTLs is infrequent. PMID- 11395209 TI - Granzyme B: a marker of risk for influenza in institutionalized older adults. AB - Risk for influenza increases with age while cellular immune responses decline. This was a prospective study to determine the relationship between cytokine and granzyme B levels in peripheral blood mononuclear cells stimulated with live influenza virus, and subsequent influenza illness. Granzyme B levels were lower in the group who later developed symptomatic laboratory-confirmed influenza (n=10) compared to the group who did not (n=90) (ANOVA, P=0.024). In contrast, none of the cytokine levels were related to the development of influenza. Thus, granzyme B is a potential marker of influenza risk in older adults. PMID- 11395210 TI - Vaccination of pigs with a recombinant porcine adenovirus expressing the gD gene from pseudorabies virus. AB - Five week old, commercially available large white pigs were vaccinated with either a single dose or two doses of a recombinant porcine adenovirus expressing the glycoprotein D gene from pseudorabies virus (PRV). Pigs were monitored for the development of serum neutralizing antibodies to PRV and challenged 3 weeks after final vaccination. Prior to challenge, pigs given 2 doses of the vaccine demonstrated boosted levels of antibody compared with those given a single dose, and all surviving pigs had increased neutralization titres over pre-challenge levels. Following challenge, pigs were monitored for clinical signs of disease, with blood and nasal swabs collected for virus isolation. All control animals became sick with elevated temperatures for 6 days post challenge, whereas; vaccinated animals displayed an increase in body temperature for only 2-3 days. Control pigs and those given a single dose all lost condition, but the group given 2 doses remained healthy. At postmortem, gross lesions of pneumonia only occurred in control animals and those given a single dose of vaccine. Histology carried out on the brains of all animals demonstrated a difference in severity of infection and frequency of immunohistochemical antigen detection between test animals, with control and single dose groups being most severely affected and pigs given 2 doses the least. Virus isolation studies demonstrated that no viraemia could be detected, but virus was found in nasal swabs from some animals in both groups of vaccinates following challenge. PMID- 11395211 TI - Mucosal immunization of mice using CpG DNA and/or mutants of the heat-labile enterotoxin of Escherichia coli as adjuvants. AB - Cholera toxin (CT) and the Escherichia coli heat-labile enterotoxin (LT) are potent mucosal adjuvants in animals associated, at least in part, with their ability to induce cAMP. While toxicity generally precludes their use in humans, a number of different subunit or genetically detoxified mutants of CT and LT have been developed. Another type of adjuvant that has been shown to be effective at mucosal surfaces comprises synthetic oligodeoxynucleotides (ODN) containing immunostimulatory CpG motifs (CpG ODN). We have previously demonstrated a synergy between CpG ODN and native toxins after intranasal (IN) administration to mice, and herein have examined whether this synergy is linked to the cAMP activity. The adjuvanticity of CpG ODN was evaluated with IN and oral delivery of tetanus toxoid or the hepatitis B surface antigen, relative to and in combination with native LT holotoxin (LTh), three active site mutants (LTS61F, LTA69G, LTE112K), a protease site mutant (LTR192G), and the B subunit of LT (LTB). At an equivalent dose, the adjuvants could generally be divided into two groups: one that included CpG ODN, LTh, LTR192G, and LTA69G which acted as strong adjuvants; and the second which comprised LTB, LTS61F, and LTE112K, which produced significantly weaker immune responses. When CpG ODN was co-administered with bacterial toxin derivatives, in most cases, no synergy between CpG and the LT derivatives was found for strength of the humoral response. Nevertheless, for both routes and antigens, CpG ODN combined with any LT derivative induced a more Type 1-like response than LT derivative alone. These results suggest that while the synergy seen previously with native toxins may have been due in part to inherent cAMP activity, it may have also depended on the particular antigen used and the route of immunization. PMID- 11395212 TI - Contrasting Epstein-Barr virus-specific cytotoxic T cell responses to HLA A2 restricted epitopes in humans and HLA transgenic mice: implications for vaccine design. AB - This study investigates the hierarchy of cytotoxic T cell (CTL) responses to twelve HLA A2-restricted epitopes from the latent, lytic and structural proteins of Epstein-Barr virus (EBV) in acute infectious mononucleosis and in healthy seropositive donors and the relative immunogenecity of these epitopes in transgenic mice. Responses to the lytic epitope were uniformly strong in all healthy seropositive individuals and acute infectious mononucleosis donors while moderate or low responses were observed to the latent and structural epitopes, respectively in both groups studied. In contrast, when HLA A2/Kb transgenic mice were immunised with these peptide epitopes, CTL responses were observed to all epitopes with a maximal response to the epitopes within the structural proteins and low to moderate responses to the latent epitopes. This hierarchy of CTL responses in mice was also reflected in an MHC stabilisation analysis. These contrasting CTL responses in humans following natural infection compared to the immunogenicity of these epitopes and their ability to stabilise MHC may need to be considered when designing an EBV vaccine. PMID- 11395213 TI - Vaxfectin enhances antigen specific antibody titers and maintains Th1 type immune responses to plasmid DNA immunization. AB - Antigen specific immune responses were characterized after intramuscular immunization of BALB/c mice with 5 antigen encoding plasmid DNAs (pDNAs) complexed with Vaxfectin, a cationic lipid formulation. Vaxfectin increased IgG titers for all of the antigens with no effect on the CTL responses to the 2 antigens for which CTL assays were performed. Both antigen specific IgG1 and IgG2a were increased, although IgG2a remained greater than IgG1. Furthermore, Vaxfectin had no effect on IFN-gamma or IL-4 production by splenocytes re stimulated with antigen, suggesting that the Th1 type responses typical of intramuscular pDNA immunization were not altered. Studies with IL-6 -/- mice suggest that the antibody enhancement is IL-6 dependent and results in a correlative increase in antigen specific antibody secreting cells. PMID- 11395214 TI - Intranasal immunogenicity of a Delta cya Delta crp-pabA mutant of Salmonella enterica serotype Typhimurium for the horse. AB - The aim of this study was to investigate the intranasal immunogenicity for the horse of a Deltacya Deltacrp-pabA mutant (MGN-707) of Salmonella enterica serotype Typhimurium (S. typhimurium). MGN-707 caused no sign of disease, was not detected in feces and a single administration induced strong Salmonella-specific serum and nasal mucosal antibody responses. All ponies had made strong salmonella specific serum IgGa, IgGb, IgA and IgM antibody responses by day 25 after the first immunization. IgM responses to salmonella lipopolysaccharide (LPS) were short lived whereas salmonella specific serum IgGa and IgGb persisted at high levels in all ponies until 83 and 140 days, respectively. Specific nasal mucosal antibody responses dominated by IgA and IgM were evident by day 25 in all ponies except one in which only specific IgGa and IgGb were evident. Specific nasal mucosal IgA persisted in most ponies until day 69. A second immunization on day 140 boosted antibody responses, and stimulated a strong nasal mucosal IgA response in the pony that failed to make an IgA response after primary immunization. At the termination of the experiment, IgA and IgGb dominated jejunal antibody responses whereas vaginal responses were mainly IgA. The latter response unequivocally confirms the existence of a common mucosal immune system in equids. The results indicate that a S. typhimurium Deltacya Deltacrp-pabA mutant has potential as an intranasal vaccine against salmonellosis in the horse. PMID- 11395217 TI - MVA: a cuckoo in the vaccine nest? PMID- 11395218 TI - Experts' opinions on the profile of optimal care for patients with diabetes mellitus type 2 in the Netherlands. AB - BACKGROUND: The St. Vincent Declaration has resulted in discussions and initiatives on optimal diabetes care during recent years. Both are based on two sources of knowledge: evidence and experience. We wanted to reveal the experience based knowledge in the Netherlands to identify essential elements or prerequisites for high quality type 2 diabetes care. METHODS: A group of 56 experts on diabetes care were invited to fill in a questionnaire. This included a ranking of 18 elements on the organization of diabetes care and 9 on patient education. RESULTS: The response rate was 87.5%. With regard to the organization of care 'active patient participation', 'protocolized care' and 'patient education' were evaluated as the most important. The integration in daily diabetes care was seen as the most important aspect of patient education. Optimal diabetes patient education would include five sessions (range: 1-10) of 1 h (range: 0.25-3) with active follow-up. The most appropriate disciplines for patient education are the diabetes nurse (chosen by 93% of the experts) and the dietician (77%). CONCLUSIONS: Optimal care for diabetes mellitus type 2 consists of structured care with integrated patient education. The majority of the experts indicated that this is not optimally organized within the Netherlands. PMID- 11395219 TI - A bleeding disorder caused by a cardiac tumor: case report. AB - A 22-year-old female developed symptomatic thrombocytopenia. On physical examination, apart from ecchymoses, a loud holosystolic murmur was heard. Echocardiography revealed a cardiac tumor. The thrombocytopenia did not respond to corticosteroids, but after surgical removal of the intracardiac tumor, a papillary fibroelastoma, the platelet count normalised. There are no similar case reports in the literature. Our case report illustrates that thrombocytopenia may be associated with a cardiac tumor and that complete physical examination is essential in every patient presenting with easy bruising. PMID- 11395220 TI - Unusual clinical presentation of a patient with multiple endocrine neoplasia type 2A. AB - The present report describes an unusual presentation of a female with MEN-2A. During 14 years she only had moderate symptoms. Occasionally she had slightly elevated basal calcitonin levels and abnormal pentagastrin tests, whereas thyroid scanning and echography were normal. At the age of 70 she developed bilateral pheochromocytoma. DNA-analysis demonstrated a germline Cys 611 Tyr mutation in the RET proto-oncogen on chromosome 10q11.2. One year after bilateral adrenalectomy again she developed overt symptoms of pheochromocytoma. The differential diagnosis and the importance of routine screening for RET mutations are discussed. PMID- 11395221 TI - Contribution of pulmonary vasoconstriction to haemodynamic instability after acute pulmonary embolism. Implications for treatment? AB - Acute pulmonary embolism with haemodynamic instability has a high mortality rate. Death results from an acute increase in right ventricular afterload, and the commonly held view is that mechanical obstruction of the pulmonary vascular bed is largely responsible for this increase. In accordance, recent treatment guidelines for severe pulmonary embolism focus exclusively on interventions aimed at relieving this mechanical obstruction, either by thrombolysis or (catheter) embolectomy. However, there is evidence to indicate that vasoconstriction is a very important contributor to the initial increase in pulmonary vascular resistance after pulmonary embolism. This is consistent with the observation that the degree of mechanical obstruction correlates at best poorly with haemodynamic manifestations. Thromboxane A(2) and serotonin are probably mainly responsible for pulmonary vasoconstriction. Cyclooxygenase inhibitors and serotonin antagonists have been shown in animal experiments to attenuate the haemodynamic response to acute pulmonary embolism and to reduce mortality. In addition, reports of a favourable response to pulmonary vasodilators in animals and in humans with acute severe pulmonary embolism have been published. In this paper, it is argued that we may need to reconsider our current therapeutic approach to patients with acute severe pulmonary embolism. Antagonising pulmonary vasoconstrictive mediators or administering pulmonary vasodilators may prove to be life-saving interventions in these patients. PMID- 11395222 TI - Benefits of antihypertensive drug treatment in elderly patients with isolated systolic hypertension. AB - Isolated systolic hypertension affects over 15% of all people older than 60 years. In the elderly, systolic hypertension is a major modifiable cardiovascular risk factor. Systolic blood pressure is associated with higher risk of an adverse outcome, whereas diastolic blood pressure is inversely correlated with total mortality, independent of systolic blood pressure, highlighting the role of pulse pressure as risk factor. Three placebo-controlled outcome trials on antihypertensive drug treatment in older patients with isolated systolic hypertension have been published: the Systolic Hypertension in the Elderly Program (SHEP), the Systolic Hypertension in Europe (Syst-Eur) Trial and the Systolic Hypertension in China (Syst-China) Trial. These three trials demonstrated the benefit of antihypertensive drug treatment. A meta-analysis was performed by pooling the patients from these three trials with a subset of patients with isolated systolic hypertension from five other trials in the elderly. Antihypertensive treatment based on a calcium-channel blocker may provide additional benefits in diabetic patients and in the prevention of dementia and renal dysfunction. The pooled results of 15693 older patients with isolated systolic hypertension prove that antihypertensive drug treatment is justified if on repeated clinic measurements systolic blood pressure is 160 mmHg or higher. PMID- 11395224 TI - Does hormonal therapy influence sexual function in men receiving 3D conformal radiation therapy for prostate cancer? AB - PURPOSE: We evaluated the effect of three-dimensional conformal radiation therapy (3D-CRT) with or without hormonal therapy (HT) on sexual function (SF) in prostate cancer patients whose SF was known before all treatment. METHODS AND MATERIALS: Between March 1996 and March 1999, 144 patients received 3D-CRT (median dose = 70.2 Gy, range 66.6-79.2 Gy) for prostate cancer and had pre- and post-therapy SF data. All SF data were obtained with the O'Leary Brief SF Inventory, a self-administered, multidimensional, validated instrument. We defined total sexual potency as erections firm enough for penetration during intercourse. Mean follow-up time was 21 months (SD +/- 11 months). The Wilcoxon signed-rank test was used to test for significance of the change from baseline. RESULTS: Before 3D-CRT, 87 (60%) of 144 men were totally potent as compared to only 47 (47%) of 101 at 1-year follow-up. Of the 60 men totally potent at baseline and followed for at least 1 year, 35 (58%) remained totally potent. These changes corresponded to a significant reduction in SF (p < 0.05). Patients who had 3D-CRT alone were more likely to be totally potent at 1 year than those receiving 3D-CRT with HT (56% vs. 31%, p = 0.012); however, they were also more likely to be potent at baseline (71% vs. 44%, p = 0.001). Although these two groups had a significant reduction in SF from baseline, their change was not significantly different from each other. CONCLUSION: These data indicate that 3D CRT causes a significant reduction in total sexual potency as compared to pretreatment baseline. The addition of HT does not appear to increase the risk of sexual dysfunction. PMID- 11395226 TI - A comprehensive review of CT-based dosimetry parameters and biochemical control in patients treated with permanent prostate brachytherapy. AB - PURPOSE: The American Brachytherapy Society recommends that postprostate implant dosimetry be performed on all patients undergoing transperineal interstitial permanent prostate brachytherapy (TIPPB) utilizing CT scan clinical target volume reconstructions. This study was undertaken to assess the recommended dosimetry parameters from a large cohort of patients undergoing TIPPB that would predict for PSA relapse-free survival (PSA-RFS). METHODS AND MATERIALS: Seven hundred nineteen consecutive patients with clinical stage T1/T2 adenocarcinoma of the prostate underwent TIPPB using either I-125 or Pd-103. Postimplant dosimetry was performed at 2 to 3 weeks with CT scan 3-dimensional reconstructions obtained on all patients. The D90 and D100 doses (defined as the minimum dose covering 90% and 100% of the prostate volume, respectively) and the V100 (defined as the percent of the prostate receiving 100% of the prescribed dose) were obtained for each patient. Regression analysis was performed on the D90 dose, D100 dose, and V100 to test for cutoff points that would predict for PSA-RFS, defined by a modification of the American Society for Therapeutic Radiology and Oncology consensus panel statement. A cutoff value was found and was subjected to subset analysis to assess for its robustness. Treatment-related factors were tested for their ability to achieve dosimetry at or above the cutoff dose. RESULTS: The median follow-up from this cohort is 30 months (7-71 months) with a 48-month PSA RFS of 89.5%. A D90 dose-response cutoff value > or =90% of the prescribed dose was identified. Prostate implants with a D90 dose <90% of the prescribed dose had an 80.4% 4-year PSA-RFS, while those with a D90 dose > or =90% of the prescribed dose had a 92.4% 4-year PSA-RFS (p = 0.001). No cutoff value was found for the V100 and D100 dose that predicted for PSA-RFS. Using the cutoff value, the D90 dose at 90% of the prescribed dose, a difference in 4-year PSA-RFS survival was identified for patients treated with I-125 (p = 0.04), Pd-103 (p = 0.01), TIPPB as monotherapy (p = 0.001), the addition of hormone therapy (p = 0.005), and TIPPB without hormone therapy (p = 0.001). The D90 dose was not significant for the group of patients treated with external beam radiotherapy and TIPPB (p = 0.15). The only significant finding from Cox regression analysis to predict for a poor D90 dose (<90% of the prescribed dose) was a CT/TRUS volume ratio >1.5 (p = 0.02). CONCLUSIONS: The American Brachytherapy Society recommends that postimplant CT-based dosimetry be performed for all patients treated with TIPPB. This prospective study identified that the D90 dose > or =90% of the prescribed dose can be used as a factor for predicting PSA-RFS in patients treated with brachytherapy. A dose-response using the D90 dose was observed for several typical clinical treatment variations used in the practice of TIPPB. Using the D90 dose appears to be a satisfactory parameter for predicting outcome in patients treated with TIPPB. PMID- 11395225 TI - A comparison of radiation dose to the bulb of the penis in men with and without prostate brachytherapy-induced erectile dysfunction. AB - PURPOSE: To retrospectively evaluate the relationship between the radiation dose to the bulb of the penis and the development of erectile dysfunction (ED) in patients undergoing permanent prostate brachytherapy without external beam radiation therapy. METHODS AND MATERIALS: Twenty-three men who developed ED after transperineal ultrasound-guided permanent prostate brachytherapy for clinical T1/T2 adenocarcinoma of the prostate gland were paired with 23 similar men who maintained potency after implantation. Potency was defined as an erection sufficient for vaginal penetration. The mean and median follow-up for the entire group was 34.6 +/- 13.7 months and 32.8 months, respectively. Patients were implanted with either (125)I (145 Gy TG-43) or (103)Pd (115 Gy, pre-NIST-99). No patient received external beam radiation therapy either before or after brachytherapy. The bulb of the penis was outlined at 0.5-cm intervals on the Day 0 postimplant CT scan. The radiation dose distribution to the bulb of the penis was defined in terms of the minimal dose delivered to 25%, 50%, 70%, 75%, 90%, and 95% of the bulb (D(25), D(50), D(70), D(75), D(90), and D(95)). RESULTS: The radiation dose delivered to the bulb of the penis in men with postbrachytherapy induced ED was statistically greater for all evaluated dosimetric parameters (D(25), D(50), D(70), D(75), D(90), and D(95)). Multivariate analysis indicated that dose to the bulb of the penis and patient age at the time of implant were predictive of postimplant ED, whereas choice of isotope had no effect. Among potent patients, 19/23 had D(50) < or = 40% of prescribed minimal peripheral dose, whereas for the impotent patients, 19/23 had D(50) >40% of the minimal peripheral dose. Of the impotent patients, 17 utilized sildenafil, with 15 experiencing a favorable response (88%). CONCLUSION: Our data suggest that prostate brachytherapy-induced impotence is highly correlated with the radiation dose delivered to the bulb of the penis. With Day 0 dosimetric evaluation, the radiation dose delivered to 50% of the bulb of the penis should be maintained at 50 Gy or less to maximize post-treatment potency. Fortunately, the majority of the brachytherapy-induced ED population responds favorably to sildenafil. PMID- 11395227 TI - PSA doubling time of prostate carcinoma managed with watchful observation alone. AB - PURPOSE: To study prostate-specific antigen (PSA) doubling time of untreated, favorable grade, prostate carcinoma. METHODS AND MATERIALS: A prospective single arm cohort study has been in progress to assess the feasibility of a watchful observation protocol with selective delayed intervention using clinical, histologic, or PSA progression as treatment indication in untreated, localized, favorable grade prostate adenocarcinoma (T1b-T2bN0 M0, Gleason Score < or = 7, and PSA < or = 15 ng/mL). Patients are conservatively managed with watchful observation alone, as long as they do not meet the arbitrarily defined disease progression criteria. Patients are followed regularly and undergo blood tests including PSA at each visit. PSA doubling time (Td) is estimated from a linear regression of ln(PSA) on time, assuming a simple exponential growth model. RESULTS: As of March 2000, 134 patients have been on the study for a minimum of 12 months (median, 24; range, 12-52) and have a median frequency of PSA measurement of 7 times (range, 3-15). Median age is 70 years. Median PSA at enrollment is 6.3 (range, 0.5-14.6). The distribution of Td is as follows: <2 years, 19 patients; 2-5 years, 46; 5-10 years, 25; 10-20 years, 11; 20-50 years, 6; > 50 years, 27. The median Td is 5.1 years. In 44 patients (33%), Td is greater than 10 years. There was no correlation between Td and patient age, clinical T stage, Gleason score, or initial PSA level. CONCLUSION: Td of untreated prostate cancer varies widely. In our cohort, 33% have Td > 10 years. Td may be a useful tool to guide treatment intervention for patients managed conservatively with watchful observation alone. PMID- 11395228 TI - Radiation dose response in patients with favorable localized prostate cancer (Stage T1-T2, biopsy Gleason < or = 6, and pretreatment prostate-specific antigen < or = 10). AB - PURPOSE: To study the radiation dose response as determined by biochemical relapse-free survival in patients with favorable localized prostate cancers, i.e., Stage T1-T2, biopsy Gleason score (bGS) < or = 6, and pretreatment prostate specific antigen (iPSA) < or = 10 ng/mL. METHODS AND MATERIALS: A total of 292 patients with favorable localized prostate cancer were treated with radiotherapy alone between 1986 and 1999. The median age was 69 years. Sixteen percent of cases (n = 46) were African-American. The distribution by clinical T stage was as follows: T1/T2A, 243 (83%); and T2B/T2C, 49 (17%). The distribution by iPSA was as follows: < or = 4 ng/mL, 49 (17%); and > 4 ng/mL, 243 (83%). The mean iPSA level was 6.2 (median, 6.4). The distribution by bGS was as follows: or = 5 in 89 cases (30%) and 6 in 203 cases (70%). The median radiation dose was 70.0 Gy (range, 63.0-78.0 Gy). Doses of < or = 70.0 Gy were delivered in 175 cases, 70.2 72.0 Gy in 24 cases, 74 Gy in 30 cases, and 78 Gy in 63 cases. For patients receiving < 72 Gy, the median dose was 68 Gy, vs. 78 Gy for patients receiving > or = 72 Gy. A conformal technique was used in 129 (44%) of cases. The median follow-up was 43 months (range, 3-153). RESULTS: For the entire cohort, the projected 5- and 8-year biochemical relapse-free survival (bRFS) rates were both 81%. For patients receiving > or = 72 Gy, the 5- and 8-year bRFS rates were both 95% vs. only 77% for patients receiving < 72 Gy, p = 0.010. For patients receiving 74 Gy, the 4-year bRFS rate was 94% vs. 96% for patients receiving 78 Gy, p = 0.90. A multivariate analysis for factors affecting bRFS rates using Cox proportional hazards was performed for all cases using the following variables: age (continuous variable), race (black vs. white), iPSA (continuous variable), bGS (< or = 5 vs. 6), Stage (T1-2A vs. T2B-C), radiation dose (continuous variable), and radiation technique (conformal vs. standard). From the multivariate analysis, only iPSA (p = 0.017, chi(2) = 5.7), and radiation dose (p = 0.021, chi(2) = 5.3) were independent predictors of outcome. Age (p = 0.94), race (p = 0.89), stage (p = 0.45), biopsy GS (p = 0.40), and radiation technique (p = 0.45) were not. CONCLUSION: There is a clear radiation dose response in patients with favorable localized prostate cancers (i.e., Stage T1-T2, biopsy Gleason score < or = 6, and iPSA < or = 10 ng/mL). At least 74 Gy should be delivered to the prostate and periprostatic tissues. With our cohort of patients, longer follow-up will be needed to assess the importance of doses exceeding 74 Gy. PMID- 11395229 TI - Can dose-response models predict reliable normal tissue complication probabilities in radical radiotherapy of urinary bladder cancer? The impact of alternative radiation tolerance models and parameters. AB - PURPOSE: To analyze the consequences of selecting alternative normal tissue complication probability (NTCP) models and parameters for evaluation of radiotherapy of urinary bladder cancer. METHODS AND MATERIALS: Treatment plans of 24 bladder cancer patients referred to radical 4-field conformal radiotherapy were analyzed. Small intestinal and rectal NTCPs were determined using both the probit and relative seriality models with several sets of published radiation tolerance parameters. Various combinations of NTCP models and parameters were applied to find the prescription dose in individual patients as well as to estimate the benefit of the conformal radiotherapy setup. RESULTS: Different risk estimates were predicted from the two NTCP models, even when the same clinical radiation tolerance doses were fitted into the two models. The demonstrated variability translated into significant deviations (7-10 Gy) in the recommended prescription doses. Even if it was possible to discriminate between a 2-field plan and the 4-field conformal setup using a given complication model and set of tolerance parameters, the estimated benefit of the conformal treatment in terms of permitted dose escalation varied with as much as 10-12 Gy between the different NTCP models/parameters used. CONCLUSION: Different NTCP models and tolerance parameters might propose different answers to important clinical questions in radiotherapy treatment of bladder cancer, such as dose prescription and scoring of rival treatment plans. We therefore recommend that the variability caused by tolerance parameter uncertainty and model selection should be taken into account in dose-response modeling of radiotherapy treatment. PMID- 11395230 TI - Radiation therapy for superficial esophageal cancer: a comparison of radiotherapy methods. AB - PURPOSE: A comparison of treatment outcomes in response to various methods of radiotherapy for superficial esophageal cancer (SEC) was carried out for a large series of patients. METHODS AND MATERIALS: During the period from March 1987 to November 1998, 147 patients with superficial esophageal cancer received definitive radiation therapy at nine radiotherapy institutions in Japan. Fifty five patients were treated with external radiation therapy alone, 69 with high dose-rate intracavitary radiation therapy with or without external radiation therapy, and 23 with low-dose-rate intracavitary radiation therapy and external radiation therapy. RESULTS: The 5-year survival rates for mucosal and submucosal cancer patients were 62% and 42%, respectively. The 5-year cause-specific survival rates for mucosal and submucosal cancer patients were 81% and 64%, respectively (p = 0.013). There was no statistically significant difference in the survival rates for either mucosal or submucosal cancer patients between treatment groups. Metastasis was observed only in submucosal cancer patients. Esophageal ulcers developed only in patients who received intracavitary radiation therapy, and were especially common in patients treated with a fraction size of 5 Gy or more. CONCLUSIONS: The use of intracavitary radiation therapy does not influence the survival or local control rate of SEC. Optimal radiotherapy methods for SEC should be determined by a randomized clinical trial. PMID- 11395231 TI - Adjuvant radiochemotherapy in the treatment of completely resected, locally advanced gastric cancer. AB - PURPOSE: To analyze the efficacy and toxicity of adjuvant whole abdomen irradiation (WAI) and concomitant chemotherapy in the treatment of completely resected, high-risk gastric cancer. METHODS AND MATERIALS: Between October 1990 and September 1997, 52 patients with completely resected gastric cancer, with lymph node and/or serosal involvement, were treated. Ages were 16-78 (median, 53.5) years. Treatment was either total- or sub-total gastrectomy, followed by WAI, 2100 cGy/21 fractions plus a 2400 cGy/16 fractions boost to the tumor bed. Chemotherapy consisted of either 5-fluorouracil (5-FU) 450-500 mg/m(2) i.v. for 5 days first and 5th week or 200-300 mg/m(2) continuous infusion during irradiation. No further chemotherapy was given. RESULTS: With a minimum follow-up of 30 months and a median follow-up of 43.5 months, 25 of the 52 patients have died. Overall 5-year survival rate is 54%. Three patients sustained Grade 3-5 complications. Two patients with Grade 5 complications (malabsorption syndrome) died 31 and 56 months after the beginning of the treatment, respectively, with no evidence of recurrent tumor. For patients with involvement of the lymph nodes alone (n = 19) the 5-year survival was 69%, which was significantly better than the 36% 5-year survival observed for those patients with both serosal and lymph node involvement (n = 26, p = 0.004). CONCLUSION: Adjuvant radiochemotherapy, WAI, and concomitant 5-FU, is a feasible and a fairly well-tolerated treatment for patients with locally advanced (involvement of the lymph nodes or serosa) gastric carcinoma who undergo complete resection. The 54% overall 5-year survival compares favorably with the survival reported after surgery alone for those patients. PMID- 11395232 TI - Effect on local control and survival of electron beam intraoperative irradiation for resectable pancreatic adenocarcinoma. AB - PURPOSE: To assess the impact on local control and survival of intraoperative radiotherapy (IORT) in resectable pancreatic adenocarcinoma. METHODS AND MATERIALS: The outcome of 127 patients surgically treated with curative intent combined with IORT was compared with the therapeutic results of 76 patients treated with surgery as exclusive treatment. RESULTS: Operative mortality and morbidity were similar in IORT and no-IORT patients. In 49 patients with locally limited disease (Stage I-II; LLD), IORT (n = 30) reduced the local failure rate and significantly prolonged time to local failure (TTLF), time to failure (TTF), and overall survival (OS) with respect to surgery alone (n = 19). The multivariate analyses, stratifying patients by age, tumor grade, resection margins, chemotherapy, and external-beam radiotherapy use, confirmed the independent impact of IORT on outcome. In patients with locally advanced disease (Stage III-IVA; LAD), IORT had an impact on local failure rate and on TTLF when combined with beam energies of greater than 6 MeV, whereas no effect on TTF and OS was observed. CONCLUSION: IORT did not increase operative mortality and morbidity and achieved a significant improvement in local control and outcome in patients with LLD. In patients with LAD, beam energies greater than 6 MeV prolonged TTLF. PMID- 11395233 TI - Decreased tumor cell proliferation as an indicator of the effect of preoperative radiotherapy of rectal cancer. AB - BACKGROUND: Rectal cancer is a common malignancy, with significant local recurrence and death rates. Preoperative radiotherapy and refined surgical technique can improve local control rates and disease-free survival. PURPOSE: To investigate the relationship between the tumor growth fraction in rectal cancer measured with Ki-67 and the outcome, with and without short-term preoperative radiotherapy. METHOD: Ki-67 (MIB-1) immunohistochemistry was used to measure tumor cell proliferation in the preoperative biopsy and the surgical specimen. MATERIALS: Specimens from 152 patients from the Southeast Swedish Health Care region were included in the Swedish rectal cancer trial 1987-1990. RESULTS: Tumors with low proliferation treated with preoperative radiotherapy had a significantly reduced recurrence rate. The influence on death from rectal cancer was shown only in the univariate analysis. Preoperative radiotherapy of tumors with high proliferation did not significantly improve local control and disease free survival. The interaction between Ki-67 status and the benefit of radiotherapy was significant for the reduced recurrence rate (p = 0.03), with a trend toward improved disease-free survival (p = 0.08). In the surgery-alone group, Ki-67 staining did not significantly correlate with local recurrence or survival rates. CONCLUSION: Many Ki-67 stained tumor cells in the preoperative biopsy predicts an increased treatment failure rate after preoperative radiotherapy of rectal cancer. PMID- 11395234 TI - Pretreatment clinical findings predict outcome for patients receiving preoperative radiation for rectal cancer. AB - BACKGROUND: As a sole modality, preoperative radiation for rectal carcinoma achieves a local control comparable to that of postoperative radiation plus chemotherapy. Although the addition of chemotherapy to preoperative treatment improves the pathologic complete response rate, there is also a substantial increase in acute and perioperative morbidity. Identification of subsets of patients who are at low or high risk for recurrence can help to optimize treatment. METHODS: During the period 1977-95, 384 patients received preoperative radiation therapy for localized adenocarcinoma of the rectum. Ages ranged from 19 to 97 years (mean 64.4), and there were 171 females. Preoperative treatment consisted of conventionally fractionated radiation to 3600-5040 cGy (median 4500 cGy) 6-8 weeks before surgery in 293 cases or low doses of <3000 cGy (median 2000 cGy) immediately before surgery in 91 cases. Concurrent preoperative chemotherapy was given to only 14 cases in this study period. Postoperative chemotherapy was delivered to 55 cases. RESULTS: Overall 93 patients have experienced recurrence (including 36 local failures). Local failures were scored if they occurred at any time, not just as first site of failure. For the group as a whole, the actuarial (Kaplan-Meier) freedom from relapse (FFR) and local control (LC) were 74% and 90% respectively at 5 years. Univariate analysis of clinical characteristics demonstrated a significant (p < 0.05) adverse effect on both LC and FFR for the following four clinical factors: (1) location <5 cm from the verge, (2) circumferential lesion, (3) near obstruction, (4) tethered or fixed tumor. Size, grade, age, gender, ultrasound stage, CEA, radiation dose, and the use of chemotherapy were not associated with outcome. Background of the surgeon was significantly associated with outcome, colorectal specialists achieving better results than nonspecialist surgeons. We assigned a clinical score of 0 to 2 on the basis of how many of the above four adverse clinical factors were present: 0 for none, 1 for one or two, 2 for three or four. This sorted outcome highly significantly (p < or = 0.002, Tarone Ware), with 5-year LC/FFR of 98%/85% (score 0), 90%/72% (score 1), and 74%/58% (score 2). The scoring system sorts the data for both subgroups of surgeons; however, there are substantial differences in LC on the basis of the surgeon's experience. For colorectal specialists (251 cases), the 5-year LC is 100%, 94%, and 78% for scores of 0, 1, and 2, respectively (p = 0.004). For the more mixed group of nonspecialist surgeons (133 cases), LC is 98%, 80%, and 65% for scores of 0, 1, and 2 (p = 0.008). In multivariate analysis, the clinical score and surgeon's background retained independent predictive value, even when pathologic stage was included. CONCLUSIONS: For many patients with rectal cancer, adjuvant treatment can be administered in a well tolerated sequential fashion-moderate doses of preoperative radiation followed by surgery followed by postoperative chemotherapy to address the risk of occult metastatic disease. A clinical scoring system has been presented here that would suggest that the local control is excellent for lesions with a score of 0 or (if the surgeon is experienced) 1, and therefore sequential treatment could be considered. Cases with a clinical score of 2 should be strongly considered for protocols evaluating more aggressive preoperative treatment, such as combined modality preoperative treatment. PMID- 11395235 TI - The impact of gap duration on local control in anal canal carcinoma treated by split-course radiotherapy and concomitant chemotherapy. AB - PURPOSE: To investigate the potential benefit of reducing the intersequence gap in patients with anal cancer treated with split-course chemoradiotherapy. METHODS: The study group consisted of 90 patients with anal squamous carcinoma treated between 1981 and 1998, using concomitant chemotherapy (CT) and radiation (RT). Median age was 65 years (range 41-87). RT was delivered in a split course, with a median gap of 37.5 days (range 4-97) between sequences. First (pelvic) sequence delivered a median dose of 40 Gy (range 36-50.4), using AP/PA megavoltage photon beams. Boost treatment (median dose 20 Gy, range 13-26) consisted of either Iridium-192 implantation (49 patients) or external beam RT (41 patients). CT consisted of 1-2 cycles of a 5-day continuous infusion of 5 fluorouracil and bolus mitomycin C, usually administered during the first week of each RT course. Median follow-up was 76.2 months. Univariate and multivariate analyses were performed to determine the factors associated with locoregional control (LRC). RESULTS: Five-year actuarial LRC was 72.5%. Factors associated with poorer LRC (univariate) were: age < or = 65, male gender, and gap > 37.5 days. Number of CT cycles (1 vs. 2 or more), boost technique (brachytherapy vs. external), and T-stage were not significantly associated with LRC. In multivariate analysis, only age (p = 0.01), and gap (p = 0.02) retained their significance. In patients older than 65 years, LRC was 92.3% and 75% for shorter and longer gaps, respectively. In younger patients, the corresponding values for LRC were 73.7% and 50%. CONCLUSION: In anal cancers, split-course RT with > 50 Gy dose delivery is difficult to avoid because of acute toxicity. The present analysis suggests that shortening the gap contributes to optimizing LRC. Gaps longer than 5 weeks correlated with poorer LRC, with especially unsatisfactory results observed in younger patients. PMID- 11395236 TI - Elective nodal irradiation in the treatment of non-small-cell lung cancer with three-dimensional conformal radiation therapy. AB - PURPOSE: Dose escalation using three-dimensional conformal radiation therapy (3D CRT) has been investigated as a means to improve local control. However, with higher doses, the risk of toxicity increases. Early in our experience, we ceased treating elective nodal areas (lymph node stations without evidence of tumor involvement) in an effort to decrease toxicity while treating the gross tumor to higher doses. This report measures the rate of regional failure without elective radiation therapy to uninvolved lymph nodes. METHODS AND MATERIALS: A total of 171 patients with non-small-cell lung cancer treated with 3D-CRT at Memorial Sloan-Kettering Cancer Center between 1991 and 1998 were reviewed. Only lymph node regions initially involved with tumor either by biopsy (55%) or radiographic criteria (node > or =15 mm in the short axis on CT) were included in the clinical target volume. Elective nodal failure was defined as a recurrence in an initially uninvolved lymph node in the absence of local failure. RESULTS: Only 11 patients (6.4%) with elective nodal failure were identified. With a median follow-up of 21 months in survivors, the 2-year actuarial rates of elective nodal control and primary tumor control were 91% and 38%, respectively. In patients who were locally controlled, the 2-year rate of elective nodal control was 85%. The median time to elective nodal failure was 4 months (range, 1-19 months). Most patients failed in multiple lymph node regions simultaneously. CONCLUSION: Local control remains one of the biggest challenges in the treatment of non-small-cell lung cancer. Most patients in our series developed local failure within 2 years of radiation therapy. The omission of elective nodal treatment did not cause a significant amount of failure in lymph node regions not included in the clinical target volume. Therefore, we will continue our policy of treating mediastinal lymph node regions only if they are clinically involved with tumor. PMID- 11395237 TI - Radical surgical resection and high-dose intraoperative radiation therapy (HDR IORT) in patients with recurrent gynecologic cancers. AB - OBJECTIVE: To determine the outcome for patients with recurrent gynecologic tumors treated with radical resection and combined high-dose intraoperative radiation therapy (HDR-IORT). METHODS AND MATERIALS: Between November 1993 and June 1998, 17 patients with recurrent gynecologic malignancies underwent radical surgical resection and high-dose-rate brachytherapy. The mean age of the study group was 49 years (range 28-72 years). The site of the primary tumor was the cervix in 9 (53%) patients, the uterus in 7 (41%) patients, and the vagina in 1 (6%) patient. The treatment for the primary disease was surgery with or without adjuvant radiation in 14 (82%) patients and definitive radiation in 3 (18%) patients. The current surgery consisted of exenterative surgery in 10 (59%) patients and tumor resection in 7 (41%) patients. Complete gross resection was achieved in 13 (76%) patients. The mean HDR-IORT dose was 14 Gy (range 12-15). Additional radiation in the form of permanent Iodine-125 implant was given to 3 of 4 patients with gross residual disease. The median peripheral dose was 140 Gy. RESULTS: With a median follow-up of 20 months (range 3-65 months), the 3-year actuarial local control (LC) rate was 67%. In patients with complete gross resection, the 3-year LC rate was 83%, compared to 25% in patients with gross residual disease, p < 0.01. The 3-year distant metastasis disease-free and overall survival rates were 54% and 54%, respectively. The complications were as follows: gastrointestinal obstruction, 4 (24%); wound complications, 4 (24%); abscesses, 3 (18%); peripheral neuropathy, 3 (18%); rectovaginal fistula, 2 (12%); and ureteral obstruction, 2 (12%). CONCLUSION: Radical surgical resection and combined IORT for patients with recurrent gynecologic tumors seems to provide a reasonable local-control rate in patients who have failed prior surgery and/or definitive radiation. Patient selection is very important, however, as only those patients with complete gross resection at completion of surgery appear to benefit most from this radical approach in the salvage setting. PMID- 11395238 TI - Xerostomia and its predictors following parotid-sparing irradiation of head-and neck cancer. AB - PURPOSE: To assess long-term xerostomia in patients receiving parotid-sparing radiation therapy (RT) for head-and-neck cancer, and to find the patient and therapy-related factors that affect its severity. PATIENTS AND METHODS: From March 1994 through January 2000, 84 patients received comprehensive bilateral neck RT using conformal and multisegmental intensity-modulated RT (IMRT) aiming to spare the major salivary glands. Before RT and periodically through 2 years after the completion of RT, salivary flow rates from each of the major salivary glands were selectively measured. At the same time intervals, each patient completed an 8-item self-reported xerostomia-specific questionnaire (XQ). To gain a relative measure of the effect of RT on the minor salivary glands, whose output could not be measured, the surfaces of the oral cavity (extending to include the surface of the base of tongue) were outlined in the planning CT scans. The mean doses to the new organ ("oral cavity") were recorded. Forty-eight patients receiving unilateral neck RT were similarly studied and served as a benchmark for comparison. Factors predicting the XQ scores were analyzed using a random-effects model. RESULTS: The XQ was found to be reliable and valid in measuring patient reported xerostomia. The spared salivary glands which had received moderate doses in the bilateral RT group recovered to their baseline salivary flow rates during the second year after RT, and the spared glands in the unilateral RT group, which had received very low doses, demonstrated increased salivary production beyond their pre-RT levels. The increase in the salivary flow rates during the second year after RT paralleled an improvement in xerostomia in both patient groups. The improvement in xerostomia was faster in the unilateral compared with the bilateral RT group, but the difference narrowed at 2 years. The major salivary gland flow rates had only a weak correlation with the xerostomia scores. Factors found to be independently associated with the xerostomia scores were the pre-RT baseline scores, the time since RT, and the mean doses to the major salivary glands (notably to the submandibular glands) and to the oral cavity. CONCLUSION: An improvement over time in xerostomia, occurring in tandem with rising salivary production from the spared major salivary glands, suggests a long-term clinical benefit from their sparing. The oral cavity mean dose, representing RT effect on the minor salivary glands, was found to be a significant, independent predictor of xerostomia. Thus, in addition to the major salivary glands, sparing the uninvolved oral cavity should be considered as a planning objective to further reduce xerostomia. PMID- 11395239 TI - Impact of hemoglobin level and use of recombinant erythropoietin on efficacy of preoperative chemoradiation therapy for squamous cell carcinoma of the oral cavity and oropharynx. AB - PURPOSE: We assessed the influence of hemoglobin level and r-HuEPO administration on response to chemoradiotherapy, locoregional tumor control, and overall survival in patients treated with neoadjuvant chemoradiotherapy and surgery for a squamous cell carcinoma of the oral cavity or oropharynx. METHODS AND MATERIALS: The 191 study patients were treated with mitomycin C (15 mg/m(2) day 1), 5 fluorouracil (750 mg/m(2)/day, days 1-5), and radiotherapy (50 Gy in 25 fractions weeks 1-5), followed by resection of the primary tumor bed and neck dissection at the General Hospital Vienna, Austria, between November 1989 and October 1998 for a T2-4, N0-3, M0 SCC of the oral cavity or oropharynx. Starting in May 1996, patients with a low hemoglobin (Hgb) before or during chemoradiotherapy received r-HuEPO 10,000 IU/kg s.c. 3-6 times/week until the week of surgery. RESULTS: On multivariate analysis, Hgb level and use of r-HuEPO were independent prognostic factors for response to chemoradiotherapy and locoregional tumor control (p < 0.01). Pathologic response to neoadjuvant therapy was also predictive of locoregional control (p < 0.001). Patients with a pretreatment Hgb > or = 14.5 g/dL had significantly higher complete response, locoregional control, and survival rates than the patients with a pretreatment Hgb < 14.5 g/dL who did not receive r-HuEPO (p < 0.05). The response, control, and survival rates in patients with a pretreatment Hgb < 14.5 g/dL given r-HuEPO were significantly higher than in low Hgb patients not given r-HuEPO (p < or = 0.001) and equivalent to patients with a pretreatment Hgb > 14.5 g/dL (p > or = 0.3). CONCLUSION: Low pretreatment Hgb is a negative prognostic factor for oral cavity and oropharyngeal SCCA patients, but was completely abrogated by r-HuEpo administration during neoadjuvant chemoradiotherapy. Randomized trials of radiation and/or chemotherapy with or without r-HuEPO for patients whose Hgb level is either low at the start of therapy or is anticipated to become low during therapy are indicated. PMID- 11395240 TI - Examining prognostic factors and patterns of failure in nasopharyngeal carcinoma following concomitant radiotherapy and chemotherapy: impact on future clinical trials. AB - PURPOSE: Concomitant chemotherapy and radiotherapy (CCRT), followed by adjuvant chemotherapy, has improved the outcome of nasopharyngeal carcinoma (NPC). However, the prognosis and patterns of failure after this combined-modality treatment are not yet clear. In this report, the prognostic factors and failure patterns we observed with CCRT may shed new light in the design of future trials. METHODS AND PATIENTS: One hundred forty-nine (149) patients with newly diagnosed and histologically proven NPC were prospectively treated with CCRT followed by adjuvant chemotherapy between April 1990 and December 1997. One hundred and thirty-three (89.3%) patients had MRI of head and neck for primary evaluation before treatment. Radiotherapy was delivered either at 2 Gy per fraction per day up to 70 Gy or 1.2 Gy per fraction, 2 fractions per day, up to 74.4 Gy. Chemotherapy consisted of cisplatin and 5-fluorouracil. According to the AJCC 1997 staging system, 32 patients were in Stage II, 53 in Stage III, and 64 in Stage IV (M0). RESULTS: Univariate analysis revealed that WHO (World Health Organization) Type II histology, T4 classification, and parapharyngeal extension were poor prognostic factors for locoregional control. Multivariate analysis revealed that T4 disease was the most important adverse factor that affects locoregional control, the risk ratio being 5.965 (p = 0.02). Univariate analysis for distant metastasis revealed that T4 and N3 classifications, serum LDH level > 410 U/L (normal range, 180-460), parapharyngeal extension, and infiltration of the clivus were significantly associated with poor prognosis. Multivariate analysis, however, revealed that T4 classification and N3 category were the only two factors that predicted distant metastasis; the risk ratios were 3.994 (p = 0.02) and 3.390 (p = 0.01), respectively. Therefore, based on the risk factor analysis, we were able to identify low-, intermediate-, and high-risk patients. Low-risk patients were those without the risk factors mentioned above. They consisted of Stage II patients with T2aN0, T1N1, and T2aN1 categories and of Stage III patients with T1N2 and T2aN2 categories. Their risk of recurrence is low (4%). Intermediate-risk patients were those with at least one univariate risk factor. They are Stage II patients with T2bN0 and T2bN1 categories and Stage III patients with T2bN2 and T3N0-2 categories. The risk of recurrence is modest (18%). High-risk patients have risk factors by multivariate analysis. They are stage T4 or N3 patients. Their risk of recurrence is high (36%). CONCLUSION: Low risk patients have an excellent outcome. Future trials should focus on reducing treatment-associated toxicities and complications and reevaluate the benefit of sequential adjuvant chemotherapy. The recurrence in treatment of intermediate risk patients is modest; CCRT and adjuvant chemotherapy may be the best standard for them. Patients with T4 and N3 disease have poorer prognosis. Hyperfractionated radiotherapy may be considered for the T4 patients. Future study in these high-risk patients should also address the problem of distant spread of the disease. PMID- 11395241 TI - Cervical lymph node metastases from occult squamous cell carcinoma: cut down a tree to get an apple? AB - PURPOSE: To review the value of extended diagnostic work-up procedures and to compare the results of comprehensive or volume-restricted radiotherapy in patients presenting with cervical lymph node metastases from clinically undetectable squamous cell carcinoma. METHODS AND MATERIALS: A systematic review was undertaken of published papers up to May 2000. RESULTS: Positron emission tomography (PET) has an overall staging accuracy of 69%, with a positive predictive value of 56% and negative predictive value of 86%. With negative routine clinical examination and computerized tomography (CT) or magnetic resonance imaging (MRI), PET detected primary tumors in 5-25% of patients, whereas ipsilateral tonsillectomy discovered carcinoma in about 25% of patients. Laser-induced fluorescence imaging with panendoscopy and directed biopsies showed some encouraging preliminary results and warrants further study. All together, the reported mucosal carcinoma emergence rates were 2-13% (median, 9.5%) after comprehensive radiotherapy and 5-44% (median, 8%) after unilateral neck irradiation. The corresponding nodal relapse rates were 8-45% (median, 19%) and 31-63% (median, 51.5%), and 5-year survival rates were 34-63% (median, 50%) and 22-41% (median, 36.5%), respectively. Retrospective single-institution comparisons between comprehensive and unilateral neck radiotherapy did not show apparent differences in outcome. Prognostic determinants for survival are the N stage, number of nodes, extracapsular extension, and histologic grade. No data were found to support the benefit of chemotherapy in this disease. CONCLUSION: Physical examination, CT or MRI, and panendoscopy with biopsies remain the standard work-up for these patients. Routine use of PET or laser-induced fluorescence imaging cannot be firmly advocated based on presently available data. Although combination of nodal dissection with comprehensive radiotherapy yielded most favorable results, its impact on the quality of life should be recognized, and the confounding effects of patient selection for various treatment modalities on therapeutic outcome cannot be ruled out. A randomized trial comparing the therapeutic value of comprehensive vs. volume-limited radiotherapy is being considered. PMID- 11395242 TI - The influence of pathologic tumor characteristics on locoregional recurrence rates following mastectomy. AB - PURPOSE: The objective of this study was to evaluate the influence of pathologic factors other than tumor size and number of involved axillary nodes on the risk of locoregional recurrence (LRR) following mastectomy. PATIENTS AND METHODS: We reviewed the medical records of 1031 patients treated with mastectomy and doxorubicin-based chemotherapy without radiation on 5 prospective clinical trials. Median follow-up was 116 months (range, 6-262 months). RESULTS: Patients with gross multicentric disease were at increased risk of LRR (37% at 10 years). However, patients with multifocal disease and those with microscopic multicentric disease did not experience higher rates of LRR than those with single lesions (17% at 10 years). Patients with lymph-vascular space invasion (LVSI) or involvement of the skin or nipple also experienced high rates of LRR (25%, 32%, and 50%, respectively). The presence of close (<5 mm) or positive margins was associated with an increased risk of LRR (45%). The increased risk of LRR observed for patients with pectoral fascial invasion (33%) was not reduced when negative deep margins were obtained. On multivariate analysis, the presence of 4 or more involved axillary nodes, tumor size of greater than 5 cm, close or positive surgical margins, and gross multicentric disease were found to be independent predictors of LRR (all, p < 0.01). In a separate analysis including only patients with 1-3 involved axillary nodes, microscopic invasion of the skin or nipple, pectoral fascial invasion, and the presence of close or positive margins were significant predictors of LRR. CONCLUSION: In addition to the extent of primary and nodal disease, other factors that predict for high rates of LRR include the presence of LVSI, involvement of the skin, nipple or pectoral fascia, close or positive margins, or gross multicentric disease. These factors predict for high LRR rates regardless of the number of involved axillary nodes. PMID- 11395243 TI - Radiation therapy after a partial response to CHOP chemotherapy for aggressive lymphomas. AB - PURPOSE: To analyze the results with involved-field radiotherapy after aggressive lymphomas had decreased in size by 50-99% in response to cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP)-based chemotherapy. METHODS AND MATERIALS: From 1988 through 1996, 294 previously untreated patients with Working Formulation intermediate-grade or large-cell immunoblastic lymphomas underwent CHOP-based chemotherapy on 2 consecutive protocols at the M. D. Anderson Cancer Center. Forty-four (15%) of these patients achieved, based on international working group guidelines, a partial (50-75%) response (n = 25), or unconfirmed complete (76-99%) response (n = 19) to a median of 6 cycles of chemotherapy. These patients were treated with salvage involved-field radiotherapy (n = 32) or chemotherapy (n = 12), e.g., MINE-ESHAP, without autologous stem-cell rescue (ASCR). RESULTS: Median follow-up was 43 months. Partial responders experienced similar outcomes to unconfirmed complete responders. Local control (4-year rates: 86% vs. 53%, p = 0.009) and progression-free survival (4-year rates: 67% vs. 8%, p < 0.0001), but not overall survival (4-year rates: 70% vs. 50%, p = 0.067) were significantly better in those who received salvage radiotherapy, which was well tolerated. CONCLUSION: Progression-free and overall survival in aggressive lymphoma patients who underwent salvage radiotherapy were similar to results reported for high-dose chemotherapy with ASCR. The role of salvage radiotherapy in partial and unconfirmed complete responders to CHOP chemotherapy justifies examination in a large, cooperative group trial. PMID- 11395244 TI - Intensity-modulated stereotactic radiosurgery using dynamic micro-multileaf collimation. AB - PURPOSE: The implementation of dynamic leaf motion on a micro-multileaf collimator system provides the capability for intensity-modulated stereotactic radiosurgery (IMSRS), and the consequent potential for improved dose distributions for irregularly shaped tumor volumes adjacent to critical organs. This study explores the use of IMSRS to provide improved tumor coverage and normal tissue sparing for small cranial tumors relative to plans based on multiple fixed uniform-intensity beams or traditional circular collimator arc based stereotactic techniques. METHODS AND MATERIALS: Four patient cases involving small brain lesions are presented and analyzed. The cases were chosen to include a representative selection of target shapes, number of targets, and adjacent critical areas. Patient plans generated for these comparisons include standard arcs with multiple circular collimators, and fixed noncoplanar static fields with uniform-intensity beams and IMSRS. Parameters used for evaluation of the plans include the percentage of irradiated volume to tumor volume (PITV), normal tissue dose-volume histograms, and dose-homogeneity ratios. All IMSRS plans were computed using previously established IMRT techniques adapted for use with the BrainLAB M3 micro-multileaf collimator. The algorithms comprising the IMRT system for optimization of intensity distributions and conversion into leaf trajectories of the BrainLab M3 were developed at our institution. The ADAC Pinnacle(3) radiation treatment-planning system was used for dose calculations and for input of contours for target volumes and normal critical structures. RESULTS: For all cases, the IMSRS plans showed a high degree of conformity of the dose distribution with the target shape. The IMSRS plans provided either (1) a smaller volume of normal tissue irradiated to significant dose levels, generally taken as doses greater than 50% of the prescription, or (2) a lower dose to an important adjacent critical organ. The reduction in volume of normal tissue irradiated in the IMSRS plans ranged from 10% to 50% relative to the other arc and uniform fixed-field plans. CONCLUSION: The case studies presented for IMSRS demonstrate significant dosimetric improvements for small, irregularly shaped lesions of the brain when compared to treatments using multiple static fields or standard SRS arc techniques with circular collimators. For all cases, the IMSRS plan yielded a smaller volume of normal tissue irradiated, and/or a reduction in the volume of an adjacent critical organ (i.e., brainstem) irradiated to significant dose levels. PMID- 11395245 TI - Long-term cerebral metabolite changes on proton magnetic resonance spectroscopy in patients cured of acute lymphoblastic leukemia with previous intrathecal methotrexate and cranial irradiation prophylaxis. AB - PURPOSE: To evaluate the long-term brain metabolite changes on (1)H-MRS in acute lymphoblastic leukemia (ALL) patients who had intrathecal methotrexate (ITMTX) and cranial irradiation (CRT) for central nervous system (CNS) prophylaxis against CNS relapse. METHODS AND MATERIALS: Thirty-seven ALL patients (12 females, 25 males) with history of ITMTX and CRT for CNS prophylaxis were studied. Age ranges at the time of diagnosis and at magnetic resonance examination were 0.8-13 years and 12-27 years, respectively. The interval since diagnosis was 5.6-19 years. T2-weighted and gradient-recalled echo (GRE) magnetic resonance imaging (MRI) and proton magnetic resonance spectroscopy ((1)H-MRS) were performed to assess brain injury. RESULTS: On MRI, 3 leukoencephalopathy (LEP) and 1 infarct were detected. Twenty-two patients had evidence of hemosiderin. On (1)H-MRS no statistically significant difference in choline (Cho)/creatine (Cr) and N-acetylaspartate (NAA)/Cr was associated with LEP. A lower Cho/Cr (p = 0.006) and NAA/Cr (p = 0.078) was observed in brains with hemosiderin. Linear-regression analysis showed no statistically significant relationship between NAA/Cr or Cho/Cr with age at diagnosis, but there was a statistically significant decreasing trend of NAA/Cr and Cho/Cr with the interval since diagnosis. CONCLUSION: Long-term brain injury in ALL survivors after CNS prophylaxis with ITMTX and CRT was reflected by decreasing NAA/Cr and Cho/Cr with the interval since diagnosis. The lower Cho/Cr associated with hemosiderin but not LEP suggested a different pathophysiology for these brain lesions. PMID- 11395246 TI - Fractionated half-body irradiation (HBI) for the rapid palliation of widespread, symptomatic, metastatic bone disease: a randomized Phase III trial of the International Atomic Energy Agency (IAEA). AB - PURPOSE: To find the fastest and most effective/efficient method to economically deliver fractionated half-body irradiation (HBI) for widespread (WS), symptomatic, metastatic bone cancer. METHODS AND MATERIALS: A Phase III trial with 3 HBI arms: (Arm A) Control (15 Gy/5 fractions/5 days); (Arm B) Hyperfractionation (HF) (8 Gy/2 fractions/1 day); (Arm C) Accelerated HF (12 Gy/4 fractions/2 days). Six countries randomized 156 patients (all with WS bone metastases): 51, 56, and 49 patients to Arms A, B, and C, respectively. There were 72 (46%) breast, 50 (32%) prostate, 9 (6%) lung, and 25 (16%) miscellaneous primary tumors. Initial performance status (PS) was 1-2 in 101 (65%) and PS 3-4 in 55 (35%). The lower, upper, and middle halves of the body were treated 79, 68, and 9 times. RESULTS: Pain relief was seen in 91% of patients (45% complete [CR] and 46% partial [PR]) within 3-8 days. Overall (OS), median (MST), and pain-free (PFS) survival was 174, 150, and 122 days. Breast tumors had a higher OS (279 days) than that of other primary tumors, but when analyzed by treatment, was not significantly different than prostate tumors in Arm A. No survival differences were found in patients with PS 1-2 vs. 3-4, CR vs. PR, bone with/without visceral metastases, or by the number of metastases (< or > 15 bone lesions). Quality of life (QOL) assessed by the percent of the remaining life free of pain was 71%; furthermore significant improvements in PS, pain, and narcotic scores were seen after HBI. Toxicity was very acceptable (41% none, 50% mild/moderate, 12% severe but transitory); more was seen with upper HBI. CONCLUSION: In terms of response, time to response, OS, MST, PFS, QOL, and toxicity, schedules for Arms A and C were similar for all but prostate primaries. Schedule for Arm B, which delivered the lowest biologic dose in the shortest time, had significantly worse results in pain relief, OS, MST, PFS, and QOL. Results indicate that, for most primary tumor types (except prostate), delivering two HBI daily doses of 3 Gy in 2 consecutive days is as effective as delivering a daily dose of 3 Gy for 5 consecutive days. Thus, this is a faster and much more convenient HBI schedule for the palliation of pain in widespread cancer. PMID- 11395247 TI - Topotecan selectively enhances the radioresponse of human small-cell lung carcinoma and glioblastoma multiforme xenografts in nude mice. AB - PURPOSE: To evaluate the therapeutic efficacy of different combinations of the DNA topoisomerase I-targeting drug, topotecan (TPT), with radiation for treatment of two human tumor xenografts. METHODS AND MATERIALS: The small cell lung carcinoma 54A and glioblastoma multiforme U87 were transplanted into nude mice. Equal i.p. injections of TPT and/or equal fractions of tumor irradiation were administered daily, for 5 consecutive days. When combined, TPT was injected at different constant time intervals prior to or after each radiation fraction. The tumor growth delay and changes in skin radiation reaction by TPT were evaluated. Tumor oxygenation was measured using the Eppendorf pO(2) histography. RESULTS: The tumor growth delay induced by such chemoradiotherapy was independent of interval and sequencing of the agents for either tumor model. The efficacy of TPT alone or in combination with radiation was always dose-dependent, although of different magnitude in the two xenografts. In 54A xenografts, TPT alone induced longer growth delay, but its combined effect with radiation was not more than additive. In contrast, U87 responded less to TPT alone, however the drug and radiation interacted synergisticly in this tumor model. Using both a radiobiological approach (tumor irradiation under normoxia vs. clamp hypoxia conditions) and the polarographic electrode measurements, it was shown that TPT did not modify tumor oxygenation and, thus, unlikely modulated oxygen-related tumor radiosensitivity. In contrast to tumors, TPT virtually unchanged skin radiation reaction. CONCLUSIONS: Our data suggest that TPT, when combined with radiation treatment of tumors, provides a therapeutic gain without substantial local and systemic adverse effects. PMID- 11395248 TI - A moderate elevation of blood glucose level increases the effectiveness of thermoradiotherapy in a rat tumor model. I. Relative contributions of glucose and heating to tumor acidification. AB - PURPOSE: To establish dose-effect relationships for tumor acidification induced by heat and glucose as a basis for testing the value of adding glucose administration to combined heat and x-ray treatment at clinically achievable glucose and temperature levels. METHODS AND MATERIALS: Rhabdomyosarcoma BA1112 was grown s.c. in the upper leg of 16-20-week-old Wag/Rij rats. Animals were given 2 consecutive 100-min periods of saline (S) or glucose (G) infusion, while keeping tumor temperature at 37 degrees, 42 degrees, or 43 degrees C for 1 or 2 periods, in various combinations, each involving 6 animals. Glucose was infused i.v. as a 20% solution at 2.4-3 g/kg/h. Tumors were heated using 2,450-MHz electromagnetic radiation, and tumor pH was measured using a 0.7 mm fiberoptic probe. RESULTS: Mean overall baseline pH was 7.00 (SD 0.10). The change induced by G37G43 (i.e., glucose infusion for a full 200 min, first 100 min at 37 degrees C, final 100 min at 43 degrees C) was -0.48 +/- 0.03 (SEM) pH units, and -0.17 +/ 0.03 for S37S43. The effect of G37G42 was -0.37 +/- 0.03 pH units, compared with -0.08 +/- 0.02 for S37S42 and -0.28 +/- 0.04 for glucose alone (G37G37). Glucose was less effective when given after or fully parallel to heating: -0.21 +/- 0.02 pH units for S43G37 and -0.37 +/- 0.02 for G43G43. CONCLUSION: The glucose induced tumor pH drop is much more pronounced than that induced by heat, both of which are dose dependent. The effects of glucose and heat seem additive if heating is started when glucose-induced acidification has reached its plateau level, but the overall effect is diminished if administration is fully simultaneous or in reversed order. Schedule G37G43 is optimal with respect to tumor acidification. Its predicted superiority in thermoradiotherapy as compared with S37S42, S37S43, and G37G42 treatment regimens was confirmed in a subsequent experimental tumor control study. PMID- 11395249 TI - A moderate elevation of blood glucose level increases the effectiveness of thermoradiotherapy in a rat tumor model II. Improved tumor control at clinically achievable temperatures. AB - PURPOSE: To assess the therapeutic gain (at the TCD(50) level) that can be obtained by boosting thermoradiotherapy with intravenous glucose infusion at different temperatures. This completes our series of studies to determine the optimal conditions and the effectiveness of glucose administration at clinically achievable glucose levels and treatment temperatures. METHODS AND MATERIALS: Subcutaneous rat rhabdomyosarcoma BA1112 was irradiated with graded single doses of 300-kV X-rays (dose range 0-60 Gy). Fifteen minutes after irradiation, a 100 min intravenous infusion was started, consisting of either glucose (20% solution, 2.4-3 g/kg/h) or saline as a control. Then heat was applied to the tumors at 42 degrees C or 43 degrees C (water bath) during a subsequent 100-min period of infusion. Tumor control was scored as the absence of palpable growth at 100 days after treatment. RESULTS: Glucose infusion enhanced tumor control independent of temperature in the range 42-43 degrees C. At 42 degrees C, the TCD(50) for X irradiation decreased by 5.9 Gy (SEM 1.8 Gy), from 41.6 (1.6) to 35.7 (1.5) Gy, and at 43 degrees C from 33.3 (1.6) to 27.3 (1.5) Gy, representing a glucose enhancement ratio of approximately 1.2. At doses corresponding to the TCD(50) at either 42 or 43 degrees C, the addition of glucose increased tumor control from 50% to 70%. An enhancement ratio of 2.1 was found for the combination of irradiation, glucose infusion, and heating at 43 degrees C, with respect to irradiation alone (TCD(50) 56.3 Gy, reanalyzed earlier data). The contribution of combined heat and glucose to tumor control represented an additive effect, probably on the hypoxic cell population. CONCLUSION: Moderate glucose administration (blood concentration 300 mg/100 mL) sizably improves experimental tumor control after combined X-irradiation and hyperthermia under clinically feasible conditions. Clinical treatment should benefit from this additional modality, in particular if unsatisfactory local control rates are due to insufficient heating. The therapeutic gain has to be evaluated further in clinical studies. PMID- 11395250 TI - High linear energy transfer carbon radiation effectively kills cultured glioma cells with either mutant or wild-type p53. AB - PURPOSE: A mutation in the p53 gene is believed to play an important role in the radioresistance of many cancer cell lines. We studied cytotoxic effects of high linear energy transfer (LET) carbon beams on glioma cell lines with either mutant or wild-type p53. METHODS AND MATERIALS: Cell lines U-87 and U-138 expressing wild-type p53 and U-251 and U-373 expressing mutant p53 were used. These cells were irradiated with 290 MeV/u carbon beams generated by the Heavy Ion Medical Accelerator in the National Institute of Radiologic Science or X-rays. A standard colony-forming assay and flow cytometric detection of apoptosis were performed. Cell cycle progression and the expression of p53, p21, and bax proteins were examined. RESULTS: High LET carbon radiation was more cytotoxic than low LET X ray treatment against glioma cells. The effects of the carbon beams were not dependent on the p53 gene status but were reduced by G(1) arrest, which was independent of p21 expression. The expression of bax remained unchanged in all four cell lines. CONCLUSION: These results indicate that high LET charged particle radiation can induce cell death in glioma cells more effectively than X rays and that cell death other than p53-dependent apoptosis may participate in the cytotoxicity of heavy charged particles. Thus, it might prove to be an effective alternative radiotherapy for patients with gliomas harboring mutated p53 gene. PMID- 11395251 TI - The effect of irradiation on the biodistribution of radiolabeled pegylated liposomes. AB - PURPOSE: The effect of total-body irradiation (TBI) on the biodistribution and pharmacokinetics of (111)In-DTPA-labeled pegylated liposomes (IDLPL) was evaluated in tumor-bearing nude mice as part of an ongoing effort to develop liposome-targeted radiosensitizers. METHODS AND MATERIALS: Mice received TBI (2 Gy or 5 Gy) according to two protocols: (1) to test the effect of radiation delivered 30 min before liposome injection on the time course of IDLPL biodistribution to tumor and normal tissues over 96 h; (2) to test the effect of radiation at times ranging from 72 h to 1 h before liposome injection on tumor and normal tissue uptake of IDLPL at 24 h. Tumor and tissue/organ levels of liposome uptake were measured by dissection and quantitation in a gamma counter. RESULTS: For most tissues (tumor, liver, kidney, lung, skin, heart, and central nervous system), irradiation did not alter IDLPL biodistribution. Splenic uptake appeared to be increased by TBI, but further analysis revealed that this effect was due to reduced splenic weight in irradiated mice. IDLPL uptake was increased in the small intestine, stomach, musculoskeletal system, female reproductive tract, and adrenal glands in irradiated mice. CONCLUSION: These findings suggest that concomitant administration of liposomal radiosensitizers during radical radiotherapy is likely to be safe. However, caution should be exercised in situations in which significant volumes of small intestine or hemopoietic tissue will be irradiated. PMID- 11395252 TI - Magnetic resonance imaging system for three-dimensional conformal radiotherapy and its impact on gross tumor volume delineation of central nervous system tumors. AB - PURPOSE: We developed an MRI system for three-dimensional planning in radiotherapy. Its contribution on gross tumor volume (GTV) delineation of central nervous system (CNS) diseases was evaluated. METHODS AND MATERIALS: The MRI system, with corrected distortion, was registered on computed tomography (CT) by means of fiducial/anatomic landmarks. In 41 consecutive patients with various CNS diseases, GTVs determined by MRI/CT registration (MR/CT-GTV) and CT alone (CT GTV) were compared. Hard copies of diagnostic MRI were shown to doctors when CT GTV was determined to simulate a conventional planning situation. Multi-observer volumetric analysis was conducted, assessing interobserver deviations among four radiation oncologists and intermethodological deviations between MR/CT-GTV and CT GTV. RESULTS: Overall, the mean of geometric distortion was significantly reduced from 1.08 mm to 0.3 mm by distortion correction (p < 0.0001). The contribution of the correction was apparent at >12.0 cm radius from the center of the magnetic field. Interobserver deviation was significantly reduced by MR/CT registration (p = 0.005). The improvement was significant for acoustic neurinoma (p = 0.038), astrocytomas (p = 0.043), and lesions at the cerebellum/brainstem (p = 0.008). The regression coefficient between MR/CT-GTV and CT-GTV was <0.9 for cerebellum/brainstem lesions, suggesting that MRI/CT-GTV was smaller than CT-GTV. CONCLUSIONS: This system is feasible for three-dimensional planning and was shown to reduce interobserver deviations in GTV delineation for CNS diseases. PMID- 11395253 TI - Technique charts for EC film: direct optical measurements to account for the effects of X-ray scatter. AB - PURPOSE: To develop a method of measuring technique charts for enhanced contrast (EC) film, to demonstrate how X-ray scatter changes the response of EC film, and to generate technique charts for general use. METHODS AND MATERIALS: We have developed a "digital cassette"-consisting of a metal plate/phosphor screen, a light guide, a photodiode sensor, and an electrometer-that can be used to measure the light generated in the phosphor screen of the film cassette. In turn, these measurements can be used to generate technique charts for EC film. The digital cassette has been used to measure technique charts for 4-MV and 6-MV X-ray beams for a variety of different phantom thicknesses, field sizes, and phantom-to cassette air gaps. RESULTS AND DISCUSSION: We have observed that the signals generated in an ionization chamber located 9.4 cm behind a 30-cm-thick water equivalent phantom increase by a factor of 1.9 when the field size is increased from 4 x 4 cm(2) to 40 x 40 cm(2) when irradiated by a 6-MV X-ray beam. However, the change in EC film response is a factor of 3.5 under the same conditions. Irradiations to optimally expose the EC film predicted by the digital cassette differ by up to 82% compared to those predicted by ion chamber measurements. Nevertheless, the technique charts measured using the digital cassette predict the response of the EC film to +/- 0.2 optical density. The overresponse of the EC film is most likely due to low-energy scattered photons, which interact with the high atomic number (Z = 64) phosphor screen of the enhanced contrast localization cassette. Therefore, simple solutions, such as placing a high atomic number material above the enhanced contrast localization cassette, can reduce this contribution by scattered photons to the signal generated in the cassettes. CONCLUSIONS: We have developed a digital cassette that can make more accurate measurements of the technique charts for EC films. Our measurements show that under some conditions, X-ray scatter can generate a large fraction of the signals recorded by the EC film. Technique charts have been generated at 4 MV and 6 MV, and these charts should have universal applicability. PMID- 11395255 TI - Turkish folk medicinal plants, Part IV: Gonen (Balikesir). AB - Eighty-four folk medicinal plants from Gonen (Turkey) are reported. Among them 73 species are wild and 11 species are cultivated plants. The folk medicinal plants are mostly used for the treatment of hemorrhoids, rheumatism, stomach and kidney ailments. PMID- 11395256 TI - Antinociceptive activity of Zizyphus spina-christi root bark extract. AB - The antinociceptive effect of the aqueous extract of Zizyphus spina-christi root bark was investigated in mice and rats. Acetic acid-induced writhing, formalin and thermal (hot plate) tests were used. The extract (50 and 100 mg/kg, i.p.) showed a dose-dependent analgesic effect in all the tests used. Its i.p. LD50 in mice was 2236.07 mg/kg. PMID- 11395257 TI - Contribution to the inventory of medicinal plants from the Bushi area, South Kivu Province, Democratic Republic of Congo. AB - The traditional phytotherapy of the lay population from the Bushi area has been surveyed from 1980 to 1990. A new record of 170 medicinal plants species representing 139 genera and 68 families has been made. Details regarding the preparation and administration of plant drugs are given. PMID- 11395258 TI - Anti-inflammatory activity of Syzygium cumini bark. AB - The ethanolic extract of the bark of Syzygium cumini was investigated for its anti-inflammatory activity in animal models. The extract did not show any sign of toxicity up to a dose of 10.125 g/kg, p.o. in mice. Significant anti-inflammatory activity was observed in carrageenin (acute), kaolin-carrageenin (subacute), formaldehyde (subacute)-induced paw oedema and cotton pellet granuloma (chronic) tests in rats. The extract did not induce any gastric lesion in both acute and chronic ulcerogenic tests in rats. Thus, the present study demonstrated that S. cumini bark extract has a potent anti-inflammatory action against different phases of inflammation without any side effect on gastric mucosa. PMID- 11395260 TI - Ursene type nortriterpene from Debregeasia salicifolia. AB - A new triterpene, 3beta-19alpha-dihydroxy-30-norurs-12-ene (1), has been isolated from the methanolic extract of Debregeasia salicifolia stems. Its structure has been elucidated with the help of one- and two-dimensional NMR and NOE studies. Lupeol, beta-sitosterol, stigmasterol and oleanolic acid are also reported for the first time from this species. PMID- 11395259 TI - Furanocoumarins from the aerial parts of Dorstenia contrajerva. AB - A new glycosylated furanocoumarin, alpha-L-rhamnopyranosyl-(1-->6)-beta-D glucopyranosyl-bergaptol (1), has been isolated from Dorstenia contrajerva together with three known furanocoumarins, catechin and epicatechin. Their structures were established using high field 2D NMR techniques. PMID- 11395261 TI - Diterpenoid and limonoids from the stem of Pterorhachis zenkeri. AB - One new diterpenoid, methyl 3alpha-hydroxy-7-oxo-dehydroabietate (1), two new limonoids, 3alpha-deacetyl-amoorastatin (2) and 9beta-amoorastatin (3), and the known limonoid amoorastatin (4) were isolated from the stem of Pterorhachis zenkeri. PMID- 11395262 TI - Shikonin production by p-fluorophenylalanine resistant cells of Lithospermum erythrorhizon. AB - Studies were conducted with a BK-39 callus culture of Lithospermum erythrorhizon, which produced seven shikonin derivatives (acetylshikonin, propionylshikonin, isobutyrylshikonin, beta,beta-dimethylacrylshikonin, isovalerylshikonin, beta hydroxyisovalerylshikonin and alpha-methyl-n-butyrylshikonin). A selection of cell aggregates of BK-39 culture on a medium containing p-fluorophenylalanine (PFP) yields a cell line possessing a higher resistance to the inhibitor than the initial culture. Selected BK-39F cultures produced almost the same profile of shikonin naphthoquinones as the initial culture. The shikonin derivative content of PFP-resistant culture was approximately two times higher than that of the control, reaching 12.6% of DW cell biomass. PMID- 11395263 TI - Cardioprotective effects of Picrorrhiza kurroa against isoproterenol-induced myocardial stress in rats. AB - The cardioprotective effect of the ethanol extract of Picrorrhiza kurroa rhizomes and roots (PK) on isoproterenol-induced myocardial infarction in rats with respect to lipid metabolism in serum and heart tissue has been investigated. Oral pre-treatment with PK (80 mg kg(-1) day(-1) for 15 days) significantly prevented the isoproterenol-induced myocardial infarction and maintained the rats at near normal status. PMID- 11395264 TI - Antibacterial activity of Picrasma javanica. AB - The methanol extracts of Picrasma javanica, leaves, seeds, stem and root barks were partitioned (petrol, dichloromethane, ethyl acetate, butanol). All obtained extracts and fractions showed a broad spectrum of antibacterial activity, while none was active against the tested moulds. PMID- 11395266 TI - Antibacterial activity and cytotoxicity of Nyctanthes arbor-tristis flowers. AB - The flowers of Nyctanthes arbor-tristis showed interesting antibacterial activity against some gram-positive and gram-negative microorganisms (chloroform and ethyl acetate extracts) and significant cytotoxic activity (petroleum ether, chloroform and ethyl acetate extracts). PMID- 11395265 TI - Antibacterial activity of luteoforol from Bridelia crenulata. AB - The methanol extract of Bridelia crenulata stem bark (50-1.5625 mg/ml) and its isolated luteoforol (2-0.25 mg/ml) showed concentration-dependent inhibiting activity against all 10 tested bacteria. PMID- 11395267 TI - Antimicrobial activity of Newtonia hildebrandtii. AB - Successive petroleum ether, dichloromethane and methanol extracts of Newtonia hildebrandtii stem bark were tested in vitro for their antifungal and antibacterial activity. The methanol extract was found to be the most effective against the tested pathogens. PMID- 11395268 TI - Antibacterial activity of the resinous exudates from Haplopappus uncinatus and Haplopappus foliosus. AB - The antibacterial activities of the resinous exudates from Haplopappus uncinatus and H. foliosus are reported. The results provide a justification to the traditional use of the resinous exudates of Haplopappus spp as an antiseptic. PMID- 11395269 TI - Anthelmintic activity of Butea monosperma. AB - The methanol extract of Butea monosperma seeds, tested in vitro, showed significant anthelmintic activity. PMID- 11395270 TI - Antimicrobial activity of Horsfieldia helwigii and Melia azedarach. AB - The methanol extracts of leaves, root and stem barks of Horsfieldia helwigii and Melia azedarach showed a broad spectrum of antibacterial activity. The activity was increased on fractionation (petrol, dichloromethane, ethyl acetate), particularly in the petrol fraction of the leaves of H. helwigii and dichlorometane fraction of the stem bark of M. azedarach. No activity was shown against tested moulds. PMID- 11395271 TI - Antimicrobial activity and cytotoxicity of Zanthoxylum budrunga. AB - The petroleum ether, chloroform and methanol extracts of the leaves and barks of Zanthoxylum budrunga have been evaluated for their antibacterial, antifungal and cytotoxic properties. PMID- 11395272 TI - Antimicrobial activity of Plicosepalus acaciae. AB - The antimicrobial activities of chloroform, methanol and aqueous extracts of Plicosepalus acaciae leaves and stem are reported. In particular, the leaf methanol extract, showing the highest level of activity against the tested standard microorganisms, was effective also against a range of gram-positive and gram-negative clinical isolates from Sudanese patients. PMID- 11395273 TI - Triterpenoid acids from Schisandra propinqua with cytotoxic effect on rat luteal cells and human decidual cells in vitro. AB - Three triterpenoid acids, nigranoic acid (1), manwuweizic acid (2), schisandronic acid (3), and other four compounds were isolated from the stems of Schisandra propinqua. Compounds 1 and 2 showed significant cytotoxic effect against human decidual cells and rat luteal cells in vitro. PMID- 11395274 TI - Cycloabyssinone, a new cycloterpene from Harrisonia abyssinica. AB - A new cycloterpene, 4beta,14-dimethyl-9,19-cyclo-5alpha,9beta-ergost-24(28)-en-3 one (cycloabyssinone) (1), and a known terpene, 3-friedelanone, were isolated from stem bark of Harrisonia abyssinica and identified by spectroscopic methods. PMID- 11395275 TI - 3,4-seco-Lupanes and other constituents from Platypodium elegans. AB - The isolation and NMR data of seco-lupane triterpenes and coumarins from Platypodium elegans are reported. PMID- 11395276 TI - Alkaloids from Cyrtanthus elatus. AB - The alcoholic extract of the fresh bulbs of Cyrtanthus elatus yielded zephyranthine (1) and 1,2-O-diacetylzephyranthine (2), together with three other known alkaloids. Complete assignment of 1H and 13C NMR spectra of compounds 1 and 2 was done by employment of two-dimensional NMR techniques. PMID- 11395277 TI - Constituents from the leaves of Rhododendron latoucheae. AB - Twelve constituents from Rhododendron latoucheae were isolated. Among them, compounds 1 and 2, named rhodolatouside A and B, respectively, are new iridoids. PMID- 11395278 TI - Biflavonoids from Ouratea multiflora. AB - A new flavone dimer, 3-hydroxy-4',5,7-trimethoxyflavone-(6-->8")-3"-hydroxy 3"',4"', 5",7"-tetramethoxyflavone, together with amenthoflavone, have been isolated from the leaves of Ouratea multiflora. Its structure was established by spectroscopic methods, including two-dimensional NMR spectroscopy. PMID- 11395279 TI - Papyriflavonol A, a new prenylated flavonol from Broussonetia papyrifera. AB - A new prenylated flavonol, papyriflavonol A, was isolated from the root barks of Broussonetia papyrifera. The structure of this compound was elucidated as 5,7,3',4'-tetrahydroxy-6,5'-di-(gamma,gamma-dimethylallyl)-flavonol (1) by spectroscopic analysis. PMID- 11395280 TI - A new cardenolide from the roots of Terminalia arjuna. AB - A new cardenolide, 16,17-dihydroneridienone 3-O-beta-D-glucopyranosyl-(1-->6)-O beta-D-galactopyranoside (1), was isolated from the roots of Terminalia arjuna. PMID- 11395281 TI - A new phenolic compound from Nigella damascena seeds. AB - A new phenolic ester has been isolated from the seeds of Nigella damascena and the structure was established as 1-O-(2,4-dihydroxy)phenylacetyl glycerol (1) by 1H and 13C-NMR spectral data and EI-MS analysis. PMID- 11395282 TI - Editorial note. 'Nocturnal paroxysmal dystonia related to a prerolandic dysplasia', Epilepsy Research 2001;43:1-9. PMID- 11395283 TI - Nitric oxide metabolites, nitrates and nitrites in the cerebrospinal fluid in children with west syndrome. AB - Nitric oxide (NO) has been implicated in the mediation of the neuronal excitotoxic cascade. In order to estimate brain NO production, cerebrospinal fluid (CSF) levels of NO metabolites, nitrates and nitrites (NN(x)) were measured in 31 children with west syndrome (WS) and in 12 controls. There was no age related change in the NN(x) levels during the first year of life. The mean of the NN(x) levels was significantly higher in patients with WS than in controls (8.43 vs. 5.27 microM; P=0.01). Analysis of the etiological subgroups showed that the patients with a symptomatic etiology of WS had significantly higher NN(x) levels than controls (P<0.005) or than the patients with a cryptogenic etiology. The cryptogenic cases, in turn, did not differ from the controls (P=0.48). Levels of NN(x) were also significantly higher in children with focal brain abnormalities (infarction, atrophy or previous infection) than in those with other abnormalities or with normal neuroradiological findings (P<0.005). No correlation was found between the NN(x) levels and the duration of the symptoms, while paired samples obtained from eight children with WS showed that the NN(x) levels rose significantly (P=0.02) within the first 40 days of symptoms. The levels of NN(x) did not correlate with the CSF levels of neuronal growth factor or with the later decline in mental performance. This study demonstrates that the production of NO can be measured in human epileptic conditions and supports the idea gained from experimental studies that NO is involved in the pathophysiology of epilepsy. However, normal levels of NN(x) in patients with cryptogenic infantile spasms suggest that an increase in NO production be due to the concomitant neuronal damage rather than seizure activity per se. The findings suggest that there are no age-related changes in the NN(x) levels during the first year of life, and that children with symptomatic WS have elevated levels of NN(x), which rise during the first 40 days of symptoms. Although the NN(x) levels cannot be used to estimate the duration of symptoms or to predict the prognosis of mental development, they may support the differentiation of symptomatic from cryptogenic etiologies of WS. PMID- 11395284 TI - Involvement of cAMP- and Ca(2+)/calmodulin-dependent neuronal protein phosphorylation in mechanisms underlying genetic predisposition to audiogenic seizures in rats. AB - It was shown that increased excitability in neurons underlying epilepsies would be maintained by abnormalities in protein phosphorylation systems. This study was initiated to compare the functioning of Ca(2+)/calmodulin- and cAMP-dependent systems of protein phosphorylation in homogenates of neocortex and hippocampus in three animal groups: genetically prone to audiogenic seizures (GPAS) rats, GPAS rats exposed to daily repeated audiogenic seizures (AGPAS rats) and nonepileptic Wistar ones. We found significant differences in phosphorylation of 270, 58, 54 and 42 kDa proteins in neocortex and hippocampus of GPAS rats in comparison with Wistar ones. Daily repeated seizures induced further modifications of phosphorylation of these proteins in only hippocampus of AGPAS rats as compared with GPAS ones. Ca(2+)-independent, functional CAMKII activity was considerably increased in hippocampus but decreased in neocortex of GPAS rats in comparison with Wistar ones. The activity of PKA was increased both in neocortex and hippocampus of GPAS rats. Daily repeated audiogenic seizures induced the decrease of Ca(2+)-independent CAMKII activity in hippocampus and the increase of PKA activity in neocortex of AGPAS rats in comparison with GPAS ones. The present results indicate that modification of 270, 58, 54, and 42 kDa proteins phosphorylation as well as altered CAMKII and PKA activities might be involved in mechanisms of genetic predisposition to audiogenic seizures. PMID- 11395285 TI - Malignant migrating partial seizures in infancy. AB - A previously unreported epileptic condition characterised by onset before 6 months of age, nearly continuous electroencephalographic seizures involving multiple independent areas originating in both hemispheres, no identifiable cause, and poor outcome has been described by Coppola et al. We report three cases presenting the same clinical and EEG pictures. They show a peculiar epileptic condition unlike the other early epileptogenic encephalopathies, so they may represent a new infantile epileptic syndrome. PMID- 11395286 TI - Neurosurgical treatment of medically intractable status epilepticus. AB - Medically intractable status epilepticus can be defined as status epilepticus (SE) that persists or recurs despite medical treatment with intravenous agents that suppress cortical activity. We describe the successful neurosurgical treatment of three patients with medically intractable status epilepticus who responded either to focal resection, multiple subpial transection, or callosal section. The duration of medically intractable status epilepticus before surgery ranged between 23 and 42 days, and multiple medical complications occurred during the failed medical therapy. We suggest that patients with medically intractable status epilepticus who fail to respond to three courses of cerebral suppressant therapy for approximately 2 weeks be considered for surgical treatment in the absence of any known remitting etiology. Focal resection and/or subpial transection is preferred for intractable partial SE with focal electrographic changes or a focal lesion demonstrated by structural or functional neuroimaging. Corpus callosotomy may be used for patients with generalized or non-localizable intractable status epilepticus. PMID- 11395287 TI - Differential effects of NMDA antagonists microinjections into the nucleus reticularis pontis caudalis on seizures induced by pentylenetetrazol in the rat. AB - It has been shown that NMDA antagonists block the tonic but not the clonic component of seizures when they are injected in the oral region of the rat pontine reticular formation (PRF). The participation of the caudal PRF in the effects of NMDA antagonists upon the tonic and the clonic components of generalized seizures induced by pentylenetetrazol (PTZ) is unknown. The aim of the present study was to evaluate the effects of unilateral microinjections of competitive and non-competitive NMDA antagonists, 2-amino-7-phosphonoheptanoic acid (AP-7) and dizocilpine (MK-801), respectively, into the nucleus reticularis pontis caudalis of the rat PRF upon seizures induced by PTZ (70 mg/kg i.p.). MK 801 induced a dose-related decrease both in the incidence of generalized tonic clonic seizures (GTCS) and in the presence of spikes in the EEG. MK-801 also increased GTCS latency. On the contrary, AP-7 did not have effects on GTCS. Interestingly, it induced ipsilateral circling behavior. These results suggest that in the caudal region of the rat PRF only non-competitive NMDA antagonists should block the generation of tonic and clonic components of generalized seizures. PMID- 11395288 TI - Effects of chronic estradiol benzoate treatment on amygdala kindled seizures in male rats. AB - In the female species, effect of estrogens on seizure activity is well documented, but not much is known on the effect of this ubiquitous steroid hormone on the seizure activity of the male species. In the present study, fully kindled male rats were treated with various doses (10, 30 and 50 microg/kg, i.p.) of estradiol benzoate (EB) daily, and kindled seizure parameters such as seizure stage (SS), after discharge duration (ADD) and stage 5 duration (S(5)D) were recorded at various times (0.25, 3 h and every 24 h for 96 h) after the first of daily EB treatments. While the 10-microg/kg dose of EB failed to produce any significant effect, the 30-microg/kg dose induced a triphasic effect on seizure parameters. An initially rapid increment of ADD (after 0.25 h), followed by significant decrease of all parameters at 48 h and later a significant increase in S(5)D was observed 96 h after the first of daily EB treatments. The 50 microg/kg dose of EB produced almost a similar but less marked pattern of effects. Pre-treatment with a 3-mg/kg dose of tamoxifen citrate (TAM), not only blocked the EB (30 microg/kg) effects till 72 h but also reduced the ADD and S(5)D significantly after 0.25 h, when compared to its control group. While pre treatment with the 10-mg/kg dose of TAM only blocked the inhibitory effects of EB 48 h after the first of daily EB treatments. Administration of the latter dose of TAM alone induced a profile similar to EB treatment. These results may suggest that in male rats, estradiol treatment can both potentiate and attenuate kindled seizure parameters in a time dependent manner, and the stimulatory effects can not be blocked by TAM pre-treatment. PMID- 11395289 TI - The impact of affectivity dispositions, self-efficacy and locus of control on psychosocial adjustment in patients with epilepsy. AB - The main hypothesis of this study was that negative and positive affectivity, self-efficacy and health-related locus of control are important for psychosocial adjustment in patients with epilepsy. These dimensions are rarely examined directly in relation to the psychosocial adjustment in these patients. Correlations between measures of these constructs and measures of psychosocial adjustment in epilepsy were investigated. One hundred and one patients answered the Washington psychosocial seizure inventory (WPSI), the positive and negative affect schedule (PANAS-X), the multidimensional health locus of control scales (MHLC), the generalized self-efficacy scale and a scale measuring self-efficacy in epilepsy. Reliability analyses, correlational analyses and multiple stepwise regression analyses were performed. Negative affectivity (NA), positive affectivity (PA) and generalized self-efficacy showed high correlations with the WPSI scales emotional adjustment, overall psychosocial adjustment and quality of life. The epilepsy self-efficacy measures showed high, but lower correlations with the same WPSI scales. The MHLC scales showed low correlations with the WPSI scales. Multiple regression analyses showed that PA, NA and measures of self efficacy explained more than 50% of the variances on emotional adjustment, overall psychosocial functioning and quality of life. In conclusion, positive and negative affectivity and self-efficacy are important predictors of perceived emotional adjustment, psychosocial adjustment and quality of life in patients with epilepsy. NA is the best predictor, but PA and self-efficacy measures give unique predictions independent of NA. PMID- 11395290 TI - Normal growth during lamotrigine monotherapy in pediatric epilepsy patients -- a prospective evaluation of 103 children and adolescents. AB - Physical maturation is one of the essential developmental processes during childhood and adolescence, which can be adversely affected by a number of internal and external factors. An important side effect associated with the long term use of some antiepileptic drugs is change in body weight, followed by an increased risk for subsequent maturational problems in pediatric epilepsy patients. To evaluate the effect of lamotrigine on body growth in children and adolescents with epilepsy, weight, height and body mass index (BMI) values of 103 pediatric epilepsy patients (m/f ratio: 53/50) treated with lamotrigine monotherapy were prospectively evaluated for a period of 18.7+/-11.8 (range 6- 71) months. Age at therapy introduction was 6.7+/-2.7 (range 1.6-16.4) years and daily lamotrigine dose was 7.4+/-2.2 (range 3.5--14.2) mg/kg body weight (BW). Standard deviation scores (S.D.S.) at therapy initiation versus follow-up were height -- S.D.S.: 0.07+/-0.42 versus 0.08+/-0.42 (P=n.s.); weight -- S.D.S.: 0.01+/-0.44 versus -0.01+/-0.43 (P=n.s.) and BMI -- S.D.S.: -0.24+/-0.47 versus 0.25+/-0.37 (P=n.s.). Lamotrigine long-term monotherapy was associated with normal body growth in pediatric and adolescent patients with epilepsy, regardless of patient age, gender or duration of therapy. PMID- 11395291 TI - Hippocampal and amygdaloid damage in partial epilepsy: a cross-sectional MRI study of 241 patients. AB - Patients with drug-refractory temporal lobe epilepsy (TLE) often have hippocampal and amygdaloid damage. The present study investigated the factors associated with the occurrence and severity of damage in patients with partial epilepsy. Magnetic resonance imaging was used to measure the volumes of the hippocampus and the amygdala in 241 patients with different durations of epilepsy. We also investigated the association of damage with the location of seizure focus and clinical factors (age at onset of seizures, lifetime seizure number and medical history of complex febrile convulsions, intracranial infection or status epilepticus) with regression analysis. We found that high lifetime seizure number (P<0.05), history of complex febrile convulsions (P<0.01), and age < or = 5 years at the time of the first seizure (P<0.01) were significant risk factors for reduced hippocampal volume in TLE patients. The severity of amygdaloid damage did not differ between TLE patients with different durations of epilepsy or seizure frequency, but complex febrile convulsions (P<0.05) and intracranial infection (P<0.05) were associated with amygdaloid damage. In patients with extratemporal or unclassified partial epilepsy, the hippocampal and amygdaloid volumes did not differ when patients with different durations of epilepsy were compared with controls. The present findings indicate that a high seizure number, the occurrence of complex febrile convulsions, and an early onset of seizures contribute to hippocampal volume reduction in patients with TLE. The data provided have important implications with regard to early and effective management and seizure control in vulnerable patients. PMID- 11395293 TI - Modeling the severity of illness of ICU patients. PMID- 11395294 TI - Recent developments in the treatment of HIV disease. PMID- 11395292 TI - Anticonvulsant activity of omeprazole in rats. AB - Omeprazole has long been used as an effective agent to treat peptic ulcer. Recent studies have shown that in addition to inhibiting the H(+)-K(+)ATPase, it also inhibits carbonic anhydrase (CA) types I, II and IV. This led us to investigate its anticonvulsant effect in a rat model of electroconvulsion. Since other carbonic anhydrase inhibitors like acetazolamide induce tolerance upon repeated use, we tested the tolerance potential of omeprazole upon repeated administration of up to 1 week. The animals were divided into four groups receiving normal saline, omeprazole 0.5, 1 or 2 mg/kg intraperitoneally. CC(50), i.e. the threshold current inducing tonic hind limb extension in 50% of the rats was established using a technoconvulsometer which delivers currents of varying intensity via ear clip electrodes. The CC(50) was established 30 min after injection of omeprazole. In another group of rats, omeprazole 2 mg/kg was given for 6 days and the CC(50) determined on days 0, 1, 3 and 6. Also the concentration of omeprazole in the brain was determined using high performance liquid chromatography. The CC(50) in vehicle-treated rats was 98 mA, which increased to 126, 135 and 162 mA with 0.5, 1 and 2 mg/kg of omeprazole, respectively. On repeat-dose studies the CC(50) on day 0 was 96 mA, on day 1 166 mA, on day 3 129 mA and on day 6 102 mA. The average brain concentration of omeprazole was 53.2+/-6.9 ng/g of brain tissue. In conclusion, this study has shown omeprazole to be an effective anticonvulsant, but rapidly develops tolerance to its anticonvulsant action. This study can stimulate interest in the development of agents with dual function -- inhibition of CA as well as the accompanying Na(+)-K(+) ATPase -- and such agents may prove to be effective anticonvulsants without exhibiting tolerance. PMID- 11395295 TI - Renal function and renal disease in the elderly: Part II. AB - In all industrialized countries, life expectancy has risen in the past 100 years. The incidence of elderly patients reaching end-stage renal disease (ESRD) and requiring renal replacement therapy has also increased. During the past few decades, the pattern of ESRD has changed significantly with the emerging predominance of elderly patients. The causes of this phenomenon are manifold and include an increasing number of chronic diseases typical of the 'third age', such as type 2 diabetes mellitus and vascular disease. In many species, a consequence of aging includes deterioration of renal function, partly due to structural alterations, and partly as the result of a diminishing blood flow. In humans, the aging kidney is characterized by modifications resulting from organic and functional disturbances. In particular, type 2 diabetes mellitus has emerged as an important condition, the microvascular and macrovascular complications of which are a common cause of morbidity and mortality in older patients. In Part II of this review, the specific aspects of renal replacement therapy in the elderly will be discussed. PMID- 11395296 TI - Prognostic factors in intensive care. AB - Predicting the outcome of critical illness remains an evolving art despite many recent advances. This review article describes the tools currently employed, appraising each in turn. The subject is viewed from the perspective that physiological reserve and inflammatory response are the essential elements in assessing prognosis in patients with multi-organ dysfunction/failure, the most commonly encountered syndrome in intensive care practice. PMID- 11395297 TI - Diagnostic approach to hypercalcemia: relevance of parathyroid hormone and parathyroid hormone-related protein measurements. AB - Background: Parathyroid hormone (PTH) and parathyroid hormone-related protein (PTH-rP) are two potent hypercalcemic hormones that act on the same targets. Autonomous secretion of the former is involved in primary hyperparathyroidism (PHPT), whereas the latter is responsible for humoral hypercalcemia of malignancy (HHM). Methods: From 250 consecutive, hypercalcemic serum samples sent to our laboratory for assessment of intact PTH, we were able to obtain clinical information, as well as an additional plasma sample for PTH-rP measurement, in 134 patients. At the time of sampling, patients could be classified into seven groups: cancer without known bone metastases (CaNoMeta, n=36), cancer with bone metastases (CaMeta, n=9), no evidence of cancer (noEvCa, n=71), sarcoidosis (Sarc, n=3), end-stage renal disease (ESRD, n=12), vitamin D overdose (VIT-D, n=2), and hyperthyroidism (Thyr, n=1). Results: In the CaNoMeta group, 29/36 patients had elevated PTH-rP levels, 9/36 patients had inappropriately elevated PTH levels, and 5/36 had elevated levels of both hormones. In the CaMeta group, three of the nine patients had inappropriately elevated PTH levels, two of them with concomitantly elevated PTH-rP levels. In the NoEvCa group, 63/71 patients had an inappropriate elevation of PTH levels and were diagnosed as having PHPT. Four of the 71 patients had elevated levels of both PTH and PTH-rP; three of them were in poor health and died within a short period of time. All of the ESRD patients had very high PTH and normal PTH-rP levels, except for one woman with high PTH-rP and undetectable PTH levels; she died from what later turned out to be a recurrent bladder carcinoma. In the Sarc, Vit-D, and Thyr groups, both PTH and PTH-rP levels were normal. Conclusions: (1) Elevated PTH-rP levels are a common finding in cancer patients without bone metastases. Intact PTH, however, should always be measured in hypercalcemic patients with malignancy because concurrent primary hyperparathyroidism is not rare. (2) Primary hyperparathyroidism accounts for hypercalcemia in 90% of patients without evidence of cancer whose PTH-rP levels may also be found to be elevated in a few cases, even some with surgically demonstrated parathyroid adenoma. PMID- 11395298 TI - Plasma levels of soluble cellular adhesion molecules in patients with arterial hypertension. Correlations with plasma endothelin-1. AB - Background: Several reports have shown that circulating, soluble cellular adhesion molecules and endothelin-1 (ET-1) are implicated in the pathophysiological events of atherosclerosis and may reflect the endothelial dysfunction characterizing this disorder. Methods: To evaluate the expression of these factors in arterial hypertension (AH), we measured plasma levels of soluble intercellular adhesion molecule-1 (sICAM-1), soluble vascular cell adhesion molecule-1 (sVCAM-1), soluble P-selectin (sP-selectin), and ET-1 in 60 untreated patients with mild to moderate AH (hypercholesterolemic: n=31, normocholesterolemic: n=29) and 30 sex- and age-matched normocholesterolemic normotensive controls. Results: Hypertensive patients exhibited significantly higher levels of sICAM-1 (234+/-21 vs. 187+/-12 ng/ml, P<0.005), sVCAM-1 (681+/ 42 vs. 589+/-23 ng/ml, P<0.005), sP-selectin (89+/-17 vs. 55+/-11 ng/ml, P<0.01) and ET-1 (6.2+/-0.7 vs. 2.4+/-0.3 pg/ml, P<0.01) than did normotensive controls. The normocholesterolemic hypertensives had lower levels of sICAM-1, sVCAM-1 (P<0.01), sP-selectin and ET-1 (P<0.05) than hypercholesterolemic hypertensives, but higher levels than normotensive controls (P<0.05). In hypertensives, plasma ET-1 was significantly correlated with mean arterial pressure (r=0.51, P<0.03) and sICAM-1 levels (r=0.64, P<0.01). In hypercholesterolemic hypertensives, LDL cholesterol was also significantly correlated with plasma levels of sICAM-1 (r=0.53, P<0.04) and sP-selectin (r=0.41, P<0.05). Conclusions: Plasma levels of soluble cellular adhesion molecules are elevated in hypertensive patients in comparison to normotensive controls and may be related to plasma ET-1 activity. The coexistence of hypercholesterolemia may enhance the plasma soluble adhesion molecule activity induced by AH. PMID- 11395299 TI - Pleural fluid myeloperoxidase as a marker of infectious pleural effusions. AB - Background: The aim of this study was to establish the diagnostic accuracy of neutrophil markers (elastase, lysozyme, myeloperoxidase) found in pleural fluid in differentiating between infectious and non-infectious pleural effusions (PE). Methods: We studied 184 patients over 18 years of age with PE, classified as either infectious (34 complicated parapneumonic, 32 non-complicated parapneumonic, 45 tuberculous) or non-infectious (31 neoplasms and 42 undiagnosed exudates). Polymorphonuclear elastase (PMN-E) was determined using an immunoactivation method and lysozyme using a turbidimetric method. Myeloperoxidase (MPO) was measured by double antibody competitive radioimmunoassay. Receiver operating characteristic (ROC) curves were used to evaluate diagnostic accuracy. Results: Pleural fluid MPO was the biochemical marker that best differentiated between infectious and non-infectious PE. The ROC area under the curve (AUC) for myeloperoxidase was 0.86. MPO values over 550 &mgr;g/l diagnosed infectious PE with a specificity of 90.4% and a sensitivity of 77.4%. After excluding purulent parapneumonic PE, the sensitivity of a pleural MPO value >/=550 &mgr;g/l was 72.6%. Conclusions: Pleural fluid MPO was the marker that best differentiated between infectious and non-infectious PE. PMID- 11395300 TI - Trimethoprim-sulphamethoxazole as primary Pneumocystis carinii prophylaxis does not increase serum homocysteine levels in HIV-positive subjects. AB - Background: We recently observed that a short course of trimethoprim 300 mg b.i.d. in healthy volunteers can cause a substantial increase in fasting plasma homocysteine levels, up to concentrations reportedly associated with atherothrombotic complications. The purpose of this study was to determine whether primary Pneumocystis carinii prophylaxis (PCP) with trimethoprim sulphamethoxazole (TMP-SMX) adversely affects serum homocysteine levels in HIV positive patients. Methods: We studied 34 subjects [29 male, 5 female, mean age 36.8+/-7.9 (S.D.) years] with no prior AIDS-defining disease who required primary PCP prophylaxis (CD4+ T-cell count <200/mm(3)). The common dose of TMP-SMX was 80/400 mg (80 mg trimethoprim and 400 mg sulphamethoxazole) once daily. Serum total homocysteine levels were determined in four samples: two collected prior to the start of TMP-SMX and two collected on average 2.6+/-2.2 and 5.3+/-3.5 months into the first year of prophylactic therapy. Results: Mean serum homocysteine was 13.9+/-3.7 &mgr;mol/l pre-treatment and 14.4+/-5.0 &mgr;mol/l during treatment with TMP-SMX, a non-significant increase of 0.5 &mgr;mol/l (95% CI: -0.5 to +1.4, P=0.34). Folate levels were equally unaffected by TMP-SMX (13.1+/-6.5 nmol/l versus 13.3+/-5.3 nmol/l, before and during therapy, respectively). Baseline folate levels did not predict the response of homocysteine to TMP-SMX, and neither did age, gender, or serum creatinine. Conclusion: Long-term therapy with 80/400 mg TMP-SMX does not adversely affect homocysteine levels. PMID- 11395301 TI - Ambulatory electrogastrography in patients with sclerodermia, delayed gastric emptying, dyspepsia, and irritable bowel syndrome. Is there any clinical relevance? AB - Background: Changes in electrogastrographic parameters are described in patients with irritable bowel syndrome, sclerodermia, dyspepsia, and delayed gastric emptying in static measurements. However, no information is available about changes in ambulatory measurements. The objective of this study was to find parameters that discriminate between these diseases using cutaneous 24-h electrogastrography. Methods: Cutaneous 24-h electrogastrography (EGG) measurements were taken from 20 patients with dyspepsia, 10 patients with systemic sclerosis (sclerodermia, SSc), 7 patients with irritable bowel syndrome (IBS), 7 patients with delayed gastric emptying, and 10 healthy volunteers. Measurements were made using a DIGITRAPPER EGG (Synectics Medical Inc., Stockholm, Sweden) and the accompanying computerized data analysis package (ElectroGastroGram Version 6.30, Gastrosoft Inc., Synectics Medical Inc., Stockholm, Sweden). Frequency and power were compared pre- and postprandially, as well as during the entire day of measurement. Results: The 24-h measurements in healthy volunteers revealed 45.00%+/-12.12% normal values (2.4-3.7 cpm), 30.10%+/ 7.15% bradygastric values (<2.4 cpm), and 24.20%+/-7.76% tachygastric values (>3.7 cpm). There was no significant change in frequency between rest and motion, but there was a significant increase in power (P<0.05). There was significantly more bradygastria in patients with dyspepsia periprandially as well as after 24 h (P<0.01) than in healthy volunteers. The mean power of patients with dyspepsia was significantly higher than that of patients with IBS (P<0.05). Conclusion: Cutaneous 24-h-EGG may be used as an additional means of differentiating between dyspepsia and IBS. PMID- 11395302 TI - Pulmonary mass and multiple lung nodules mimicking a lung neoplasm as amiodarone induced pulmonary toxicity. AB - Amiodarone is an effective anti-arrhythmic agent. However, during long-term therapy, patients can develop severe adverse pulmonary reactions that are potentially life-threatening. A case of amiodarone-induced pulmonary toxicity is presented in a 78-year-old woman. She developed dyspnea and a pulmonary mass with associated multiple lung nodules mimicking a lung cancer following 5 years of treatment with amiodarone for atrial fibrillation. After drug withdrawal, and without any additional treatment, clinical and radiological improvement was observed, and radiological findings resolved completely within 6 months. PMID- 11395303 TI - Isolated pericardial empyema secondary to Staphylococcus aureus. AB - Pericardial empyema, or purulent pericarditis, is a condition uncommonly encountered in routine medical practice but one which is rapidly fatal if left untreated. Early diagnosis is paramount if the patient is to survive. This case report highlights the covert nature of presentation which may occur, hindering early diagnosis, and reviews the current literature. PMID- 11395304 TI - Sweet's syndrome associated with monosomy 7 myelodysplastic syndrome. AB - Acute febrile neutrophilic dermatosis (Sweet's syndrome) is a reactive skin process frequently associated with inflammatory and neoplastic diseases, but particularly with hematologic malignancies. It usually precedes the underlying disorders for months or even years. Much of the evidence for this is based on a small series of case reports and reviews of the literature. Recently, immunological theories have suggested that helper T cell type 1 is involved in the pathogenesis of Sweet's syndrome. This process causes stimulation of the cytokine cascade, which may be responsible for the local and systemic activation of neutrophils and histiocytes. Clinically, Sweet's syndrome is characterized by an acute eruption of painful erythematous or violaceous plaques or nodules with fever, malaise, neutrophilic leukocytosis, and an elevated erythrocyte sedimentation rate. Peripheral blood neutrophilia is frequent and is one of the diagnostic criteria. However, 53% of patients with Sweet's syndrome linked to hematologic malignancies do not present any neutrophilia but rather granulocytopenia. Abnormal functioning of neutrophils is possible in many diseases. We report a case of a middle-aged male patient presenting Sweet's syndrome and granulocytopenia due to myelodysplasia and an anomalous chromosome seven (7-) with poor prognosis. PMID- 11395305 TI - Internal medicine in Belgium. PMID- 11395306 TI - Corrigendum to 'Pseudo-iron deficiency in a French population living near high voltage transmission lines: a dilemma for clinicians'. PMID- 11395307 TI - Characterization of r-[11C]rolipram for PET imaging of phosphodieterase-4: in vivo binding, metabolism, and dosimetry studies in rats. AB - The high-affinity phosphodiesterase-4 (PDE4) inhibitor R-rolipram and the less potent S-enantiomer, both labeled with (11)C, were evaluated in vivo in rats. Regional brain uptake of R-[(11)C]rolipram was higher than R/S-[(11)C]rolipram, whereas S-[(11)C]rolipram retention subsided rapidly to levels below blood. Binding of R-[(11)C]rolipram was selective for PDE4 over PDE1, since treatment with PDE4 competitors Ro 20-1724, or R-, S- or R/S-rolipram, but not with the PDE1 inhibitor vinpocetine, significantly reduced radioactivity uptake to non specific levels. R-Rolipram (ED(50) congruent with 0.04 mg/Kg) was approximately 13 fold more potent than S-rolipram at inhibiting R-[(11)C]rolipram binding in all brain regions. Nevertheless, S-[(11)C]rolipram appears to be unsuitable for measuring the non-specific binding of R-[(11)C]rolipram. Only unchanged R [(11)C]rolipram was detected in rat brain homogenates. Additionally, the estimated absorbed radiation dose extrapolated to humans was low. These results support further investigation of R-[(11)C]rolipram in studying PDE4 in vivo by positron emission tomography imaging. PMID- 11395308 TI - Potential (18)F-labeled biomarkers for epidermal growth factor receptor tyrosine kinase. AB - As PET candidate tracers for EGFr-TK, five 4-(anilino)quinazoline derivatives, each fluorinated in the aniline moiety, were prepared. Each was tested in vitro for inhibition of EGFr autophosphorylation in A431 cell line. The leading compounds were then radiolabeled with (18)F and cell binding experiments, biodistribution and PET studies in A431 tumor-bearing mice were performed. Metabolic studies were carried out in a mice control group. From our results, we concluded that while in vitro experiments indicates efficacy of 4 (anilino)quinazoline compounds, kinetic factors and rapid blood clearance make them unsuitable as tracers for nuclear medicine imaging of EGFr-TK. PMID- 11395309 TI - Radiosynthesis and in vivo evaluation of the pseudopeptide delta-opioid antagonist [(125)I]ITIPP(psi). AB - The radioiodinated tetrapeptide delta-opioid antagonist [(125)I]ITIPP(psi) [H Tyr(3'I)-Ticpsi[CH2NH]Phe-Phe-OH] (Ki(delta) = 2.08 nM; Ki(micro)/Ki(delta) = 1280) has been synthesized and evaluated as a potential lung tumour imaging agent. [(125)I]ITIPP(psi) was obtained, via electrophilic iodination, in 46% yield (>44,000 MBq/micromol) from the parent TIPP(psi). The biodistribution of [(125)I]ITIPP(psi) in nu/nu mice bearing SCLC-SW210.5 xenographs revealed good uptake and prolonged retention of radioactivity in organs known to possess delta opioid receptors. Metabolite analysis showed that [(125)I]ITIPP(psi) was largely unmetabolized at 25 min PI and blocking studies showed significant reduction of uptake of the tracer in the brain, liver, intestine and tumor indicating that the iodinated tetrapeptide binds to delta opioid receptors in vivo. PMID- 11395310 TI - Comparison of in vivo dopamine D2 receptor binding of [(123)I]AIBZM and [(123)I]IBZM in rat brain. AB - [(123)I]AIBZM, (S)-5-[(123)I]-Iodo-N-[(1-ethyl-2-pyrrolidinyl)]methyl-4-amine-2 methoxybenzamide is a derivative with high affinity for the D2 receptor. Labeling was achieved by the Iodogen method. The in vivo affinity for the D2 receptor and the biological characteristics were performed in rats. The brain uptake of [(123)I]AIBZM was significantly lower, however the striatum/cerebellum ratio (2h p.i.) was higher than that of [(123)I]IBZM. Because of the high affinity and its possibly lower unspecific binding compared to [(123)I]IBZM, [(123)I]AIBZM may be a potential imaging agent for the D2 dopamine receptor. PMID- 11395311 TI - A simple and efficient in vitro method for metabolism studies of radiotracers. AB - In vitro metabolism of acetylcholinesterase inhibitors containing 3 [(18)F]fluoromethylbenzyl- ([(18)F]1) and 4-[(18)F]fluorobenzyl-piperidine moieties ([(18)F]2) was studied and compared with the in vivo metabolism. Defluorination of the [(18)F]1 mainly occurred to generate [(18)F]fluoride ion both in vitro and in vivo. In contrast, the [(18)F]2 was converted into an unknown polar metabolite in both metabolism methods and another metabolite, 4 [(18)F]fluorobenzoic acid in vitro. These results demonstrated that the in vitro method can be used to predict the in vivo metabolism of both radiotracers. PMID- 11395312 TI - Effect of administration route on FES uptake into MCF-7 tumors. AB - We have observed that intraperitoneal administration of [(18)F]fluoroestradiol (FES), a radiolabeled estrogen receptor ligand, results in higher abdominal organ uptake and slower blood clearance than intravenous administration in female mice. In SCID mice bearing MCF-7 human tumors SC, IP administration resulted in tumor uptake that was only about one third that obtained with IV administration. Thus, the route of administration of a radiopharmaceutical for imaging or radiotherapy of a tumor in the abdomen, an ovarian tumor, for example, could have a profound effect on the efficiency and selectivity of delivery of the agent to the tumor. PMID- 11395313 TI - The effect of molecular weight on nonspecific accumulation of (99m)T-labeled proteins in inflammatory foci. AB - Although several proteins have been proposed and tested for scintigraphic detection of infection, the most optimal characteristics of a protein for this application have not yet been determined. Molecular weight (MW) of the protein, its charge, shape, carbohydrate content, characteristics of the radionuclide and receptor interactions are factors that could affect the in vivo behavior of the infection imaging agent. The effect of molecular weight on nonspecific accumulation of (99m)Tc-labeled proteins in inflammatory foci was studied in a rat model. METHODS: Eleven proteins whose MWs ranged from 2.5 kDa up to 800 kDa were labeled with (99m)Tc using the hydrazinonicotinamide (HYNIC) chelator. Rats with S. aureus infection were injected i.v. with 15 MBq (99m)Tc-labeled protein. Gamma camera images were acquired and biodistribution of the radiolabel was determined ex vivo. RESULTS: From biodistribution data no significant correlation was found between abscess uptake and molecular size of the (99m)Tc-labeled proteins that were studied. Fast blood clearance with predominant uptake in liver and spleen was found for the largest proteins (MW 669 kDa-800 kDA). For proteins of intermediate size (MW 66 kDa -206 kDa) we found relatively slow blood clearance with relatively moderate uptake in liver and spleen. For smaller proteins (MW 2.5 kDa -29 kDa) rapid blood clearance with predominant kidney uptake was observed. The abscess uptake of the (99m)Tc-labeled proteins (%ID/g, 24 h p.i.) was highest for serum proteins IgG and BSA. Abscess uptake correlated well with blood levels: r = 0.95 and 0.84 at 4 and 24 h respectively (P < 0.005). The abscess-to-muscle ratios varied from 2.1 to 17.8 at 24 h p.i. with highest values for alpha-2 macroglobulin (MW 725 kDa) and the intermediate sized proteins (MW 66-206 kDa). Gamma camera imaging showed localization of all radiotracers at the site of infection with abscess-to-background ratios (A/B) ranging from 1.4 to 7.0 (IgG) at 20 h p.i. The serum proteins IgG and BSA showed highest blood levels and best infection imaging characteristics. CONCLUSION: Not molecular weight but blood residence time is the principal factor that determines localization of a nonspecific tracer protein in infectious foci. The ideal nonspecific infection imaging agent is a protein with a long circulatory half-life. From the proteins tested here IgG and albumin showed the best characteristics for an infection imaging agent. PMID- 11395314 TI - Synthesis and evaluation of a macrocyclic bifunctional chelating agent for use with bismuth radionuclides. AB - The detailed synthesis of the bifunctional chelating agent 2-(p isothiocyanatobenzyl)-1,4,7,10,13-pentaazacyclopentadecane-N,N',N",N"',N"" pentaacetic acid (BF_PEPA) is reported. This ligand was conjugated to monoclonal antibody B3 and the resultant immunoconjugate radiolabeled with (205,206)Bi. The in vivo stability of the radiolabeled immunoconjugate, and targeting characteristics were determined by biodistribution studies in A431 xenograft tumor-bearing mice sacrificed at 0.5, 1, 2, 4, and 24 hr. Results indicate that BF_PEPA appears to not be a suitable bifunctional chelating agent for sequestering isotopes of Bi(III) for radioimmunotherapy applications. PMID- 11395315 TI - Synthesis of a new bisphosphonic acid ligand (SEDP) and preparation of a (188)Re (Sn)SEDP bone seeking radiotracer. AB - The new bisphosphonate ligand SEDP (2-sulfonato-1,1-ethylidene bisphosphonic acid) has been synthesized and characterized, including the determination of the protonation constants, and used to form (188)Re-(Sn)SEDP from Na(188)ReO4. The title compound (188)Re-(Sn)SEDP shows slightly greater bone uptake and less kidney uptake than (188)Re-(Sn)HEDP in rat biodistribution studies. PMID- 11395316 TI - Synthesis, purification and biodistribution of (205)Bi-DOTMP, visualizing bone deposition patterns with autoradiography. AB - A HPLC system has been developed for carrier free and rapid delivery in a physiological buffer of the alpha-particle emitting bone-seeking radiopharmaceutical (212)Bi-DOTMP. (205)Bi-DOTMP was synthesized and HPLC purified to mimic and visualize the deposition pattern in bony tissues of (212)Bi DOTMP by autoradiography. Inhomogeneous bone deposits were found with the highest concentration in the bone matrix, the endosteum and in the growth zones of young mice. Analysis of urine samples showed that (205)Bi-DOTMP was cleared as an intact complex. PMID- 11395317 TI - Evaluation of [(99m)Tc] liposomes as lymphoscintigraphic agents: comparison with [(99m)Tc] sulfur colloid and [(99m)Tc] human serum albumin. AB - This study investigates the use of [(99m)Tc] liposomes for the detection of sentinel lymph nodes. A variety of [(99m)Tc] liposome formulations were compared with common lymphoscintigraphic agents including [(99m)Tc] regular-sulfur colloid (SC), [(99m)Tc] 0.22 microfiltered-SC, [(99m)Tc] reduced heating time 0.22 microfiltered-SC, and [(99m)Tc] human serum albumin (HSA) in rabbits. Images were acquired for the first 60 minutes and at 24 hours, followed by tissue biodistribution study. All agents except [(99m)Tc] regular SC demonstrated good migration from the injection site. Agents were retained in the popliteal node at 24 hours to varying degrees as follows: both [(99m)Tc] filtered SC preparations > [(99m)Tc] regular SC > [(99m)Tc] liposomes > [(99m)Tc] HSA. [(99m)Tc] liposome imaging can be used to develop novel liposome compositions with improved lymph node diagnostic and drug delivery characteristics. PMID- 11395318 TI - 99mTc labeled VIP analog: evaluation for imaging colorectal cancer. AB - Early and reliable diagnosis of colorectal cancer continues to be demanding and challenging. Colorectal cancer cells express Vasoactive Intestinal Peptide (VIP) receptors in high density. We have prepared a VIP analog (TP3654), labeled it with (99m)Tc, and evaluated it in experimental animals as an agent for imaging colorectal cancer. The tissue distribution of (99m)Tc-TP3654 has been compared with that of (111)In-DTPA-Octreotide and (99m)Tc-anti-CEA scan in nude mice bearing human colorectal cancer LS174T. Finally, pharmacokinetic and tissue distribution studies of (99m)Tc-TP3654 have been performed in four normal human volunteers. Data suggest that (99m)Tc-TP3654 can be prepared efficiently without loss of its receptor specificity and biological activity. Although the 24 hr tumor uptake of (99m)Tc-TP3654 in the animal model used was modest (0.21 +/- 0.07% I.D./g), the tissue distribution profile was more favorable than that of (111)In-DTPA-Octreotide or (99m)Tc-anti-CEA scan. Human studies indicated that (99m)Tc-TP3654 had no adverse effect in any subject. Within 24 hours, approximately 70% of the injected dose cleared through the kidneys, and approximately 20% through the hepatobiliary system. In these non-fasting volunteers hepatobiliary clearance was slow and in cancer patients tumor uptake was rapid. Data suggest that (99m)Tc-TP3654 is a promising agent for imaging colorectal cancer. PMID- 11395319 TI - [[(18)F]N-(4'-fluorobenzyl)-4-(3-bromophenyl) acetamide for imaging the sigma receptor status of tumors: comparison with [(18)F]FDG, and [(125)I]IUDR. AB - A series of biodistribution studies were conducted with the radiotracer, [(18)F]N (4'-fluorobenzyl)-4-(3-bromophenyl)acetamide, [(18)F]1 in nude mice bearing tumor xenografts of the mouse mammary adenocarcinoma, line 66. This radiotracer has a high affinity for both sigma(1) and sigma(2) receptors. In vivo studies were also conducted in order to assess the effect of blocking sigma(1) receptors on tumor uptake and the tumor:background ratio of this radiotracer. The results of these studies revealed that blocking the sigma(1) receptor so that only the sigma(2) receptors are labeled in vivo, results in a higher tumor:background ratio with only a small reduction in the tumor uptake of the radiotracer relative to the no carrier-added (i.e., nonselective) conditions. Comparative in vivo studies were also conducted with the anatomic and metabolic imaging agent, [(18)F]FDG, and a radiolabeled DNA precursor, [(125)I]IUdR. Both of these radiolabeled compounds represent classes of agents that have been proposed for imaging the proliferative status of solid tumors. The results of these studies indicated that a sigma(2) selective imaging agent may be, 1) a better anatomic imaging agent for breast cancer than [(18)F]FDG, and 2) a better functional imaging agent than the radiolabeled DNA precursors, [(123/124)I]IUdR and [(11)C]thymidine, for measuring the proliferative status of breast tumors with PET and SPECT. However, additional studies will be needed to compare sigma(2)-selective imaging agents with [(18)F]FLT in order to determine which is the more appropriate imaging agent for measuring the proliferative status of breast tumors with PET. PMID- 11395320 TI - Significance of (111)In-DTPA chelate in renal radioactivity levels of (111)In DTPA-conjugated peptides. AB - Metabolic studies of (111)In-DTPA-labeled polypeptides and peptides showed that the radiolabeled (poly)peptides generated (111)In-DTPA-adducts of amino acid that possess long residence times in the lysosomal compartment of the tissues where (poly)peptides accumulated. However, a recent study suggested that metal-chelate methionine (Met) might possess in vivo behaviors different from metal-chelate adducts of other amino acids. In this study, to elucidate whether some biological characteristics of Met may accelerate the renal elimination rate of (111)In-DTPA adduct of Met into urine, (111)In-DTPA-Met(1)-octreotide was synthesized and the renal handling of (111)In-DTPA-Met was investigated using (111)In-DTPA-L-Phe(1) octreotide (Phe represents phenylalanine), which was reported previously, as a reference. Both (111)In-DTPA-conjugated octreotide analogs were stable against 3 h incubation in murine serum at 37 degrees C. Both (111)In-DTPA-octreotide analogs also showed rapid clearance of the radioactivity from the blood and similar accumulation of the radioactivity in the kidney. No significant differences were observed in the renal radioactivity levels from 10 min to 24 h postinjection between the two. Metabolic studies indicated that (111)In-DTPA Met(1)-octreotide and (111)In-DTPA-L-Phe(1)-octreotide generated (111)In-DTPA adducts of Met and Phe, respectively, as the final radiometabolites at similar rates. These findings suggested that the long residence times of the radioactivity in tissues after administration of (111)In-DTPA-labeled peptides and polypeptides would be attributed to inherent characteristics of (111)In-DTPA chelate. PMID- 11395321 TI - Analysis of residual solvents in 2-[(18)F]FDG by GC. AB - A gas chromatography method has been developed for the measurement of the residual acetone, ethanol and acetonitrile in 2-deoxy-2-[(18)F] fluoro-D-glucose (2-[(18)F]FDG), in accordance with the pending FDA revision on the drug. The detections limits were 0.1 ppm for all three solvents. Good precision and linearity were obtained over ranges spanning the allowable concentration levels proposed by FDA. The amounts of the three solvents in our routine 2-[(18)F]FDG products have been found well below the maximum permissible levels. The method is very amenable to quality control testing for the radiopharmaceutical. PMID- 11395322 TI - An automated method for registering and quantifying scratching activity in mice: use for drug evaluation. AB - A method, which allows the automated registration of the scratching activity of the hind legs of a mouse for periods longer than 24 h, has been developed and validated. Aluminium rings are placed around the hind limbs of mice just above the ankle. Mice are housed individually in cages placed on the scratch detection unit (SDU), which contains ferrite rods fitted with copper coils. The movement of the metal rings in the field generated by the coils elicits a signal that can be transformed into peaks of different frequencies. Scratching was defined as a regular waveform with a frequency of > 15 Hz and with an amplitude of < 200 mV. Quantification of spontaneous scratching by chronic proliferative dermatitis mice or histamine and Compound 48/80-treated C57BL/Ka mice was performed visually (using a video recording showing both the mice and the computer-generated signal) and using the pruritus detection system (PDS = SDU plus computer program). The correlation between visual and automatic quantification of spontaneous cpdm mouse scratching was 81.7+/-2.5% (mean +/- S.E.M.). For histamine, the correlation was 85.6+/-3.6% (mean +/- S.E.M.) and for Compound 48/80, 85.8+/-3.1% (mean +/- S.E.M.). The PDS routinely detected less than 5% false-positive signals. PMID- 11395323 TI - A rapid electron-capture gas chromatographic procedure for the analysis of p hydroxymephenytoin. AB - An electron-capture gas chromatographic procedure was developed for detection and quantification of p-hydroxymephenytoin (OHMEP), a metabolite of S-mephenytoin, in human liver microsomal preparations. OHMEP was derivatized with pentafluorobenzoyl chloride (PFBC) under basic aqueous conditions prior to analysis on a gas chromatograph equipped with a capillary column and an electron capture detector. Dextrorophan was carried through the procedure as internal standard. The structure of the PFB derivative was confirmed using combined gas chromatography-mass spectrometry (GC-MS). The procedure is rapid and reproducible and produces a stable derivative that has excellent chromatographic properties. The limit of detection was less than 5 ng/ml, and the method was applied to extracts of human liver microsomes, which had been incubated with S-mephenytoin [a probe substrate for cytochrome P450 (CYP) 2C19]. PMID- 11395324 TI - Central Effects Index--a semiquantitative method for the assessment of CNS toxicity of local anaesthetic agents in sheep. AB - Local anaesthetic agents can cause central nervous system (CNS) and cardiovascular system toxicity. Whereas most previous work has described their behavioural CNS effects qualitatively (i.e., absence or presence of convulsions), we wished to describe their CNS effects more quantitatively. We hypothesised that early CNS excitatory or disinhibitory effects leading to convulsions represent a prodrome to the onset of chaotic dynamics in the form of a bifurcation in the chaotic map. We therefore used a chaotic map with a continuous scale to model their CNS effects. A Central Effects Index (CEI) was developed from our observation of behaviours before and after intravenous (iv) administration of local anaesthetic agents in graded doses to conscious sheep. These behaviours were ranked in severity, and modelled according to a logistic population growth equation using the onset of convulsive behaviour and death as point attractors. The behaviours, scaled to the maximum and area under the curve (AUC) CEI units, were then compared for intravenous doses of (+/-)-RS-bupivacaine and (-)-(S) bupivacaine (or levobupivacaine), which is being evaluated as a substitute for (+/-)-RS-bupivacaine. (-)-(S)-bupivacaine produced smaller maximum and AUC CEI values at 75 and 100 mg doses, but equivalent values at 150 and 200 mg when the doses exceeded the convulsant threshold. It was concluded that the CEI provides a useful quantitative tool for evaluating these agents in subconvulsant doses, and that the CNS stimulatory potency of (-)-(S)-bupivacaine is less than that of (+/ )-RS-bupivacaine. PMID- 11395325 TI - A standardized procedure for using human corpus cavernosum strips to evaluate drug activity. AB - The main problem of using human corpus cavernosum (HCC) tissue to perform bioassay is linked to its limited availability further complicated by the heterogeneous source of the tissues used. Here, we show that gender reassignment is a reliable source of human tissue without major ethical problems. Indeed, the entire corpus cavernosum is obtained from the surgery procedure, which allows creating a standardized procedure to prepare HCC strip. In addition, human tissue, if kept in the fridge in the condition described, does not loose its ability to contract to phenylephrine (PE; alpha agonist), angiotensin II (AG II) and KCl up to 4 days. Furthermore, once contracted with PE, HCC relaxes to acetylcholine (endothelium-dependent mechanism); sodium nitroprusside (endothelium-independent mechanism); cromakalim (CRK), a K(ATP) channel opener; or alprostadil, a synthetic PGE2 (ALPR). In conclusion, we have standardized a procedure that allows the use of HCC strips to evaluate drug activity and/or to study pathophysiological mechanisms with an intact functional human tissue up to 4 days from the surgery procedure. PMID- 11395326 TI - A new approach to evaluate toxicity of gases on mobile cells in culture. AB - A novel technique is described that measures the degree of toxicity of short-term exposure to gaseous pollutants or other chemical compounds on cultured cells, in 30 min. This technique, based on the study of the mobility properties of activated macrophages, consists of an image analysis procedure incorporating a specific exposure chamber (EC). The EC, which is developed from commercial culture flasks (50 ml, 25 cm(2) of culture surface), was first used to maintain cells in culture conditions, overnight, prior to the assay. In order to measure toxicity, it was then connected to the gaseous pollutant or chemical source. After exposing the culture medium and cells to the gas stream for 10 min, fMLP, a chemotactic factor, was added and the mobility of the macrophages measured by superimposing sequential analogue images captured by a CCD camera that were digitised and analysed using a software developed for this purpose. For example, the effect of ozone on macrophage-like cell (THP-1) was investigated. After exposure to 0.1 and 0.5 ppm, cells lost, respectively 79% and 90% of their mobility, compared to the control sample. PMID- 11395327 TI - A novel procedure for daily measurements of hemodynamical, hematological, and biochemical parameters in conscious unrestrained rats. AB - Accurate and chronic measurements of various parameters in conscious animals are fundamental for depicting pathological chronic conditions and their etiology in many experimental models, but they are often difficult to achieve. The aim of the present work was to develop and describe step-by-step a reproducible surgical procedure and daily manipulations for continuous, chronic use of conscious rats as models towards a better understanding of various cardiovascular and renal diseases and the testing of novel pharmacological drugs. The complete apparatus involved the use of a series of specialized devices (harness, rotating swivel, revolving arm) supporting a flexible, permanently implanted vascular catheter into the left femoral artery up to the abdominal aorta connected to a miniaturized individual peristaltic pump for delivering fluid at a constant rate. Such a set-up also enabled easy, quick, and reproducible daily blood sampling for the evaluation of more than 20 parameters, including the monitoring of heart rate (HR) and blood pressure in freely moving conscious rats. The overall success and survival rate reached 98% over 14 days and could be extended further. This model represents a much needed and valuable advance in surgical research techniques to evaluate the hemodynamic, hematological, biochemical, pharmacokinetic, and toxicological profile of any new drugs over time in conscious animal models such as rats. What makes this procedure satisfactory is the long-term reliable arterial access and reproducibility of the methodological approach for accurate and continuous measurements, minimizing the stress or invasiveness associated with the use of currently employed systems. PMID- 11395328 TI - Evaluation of scavenging activity assessed by Co(II)/EDTA-induced luminol chemiluminescence and DPPH* (2,2-diphenyl-1-picrylhydrazyl) free radical assay. AB - The scavenging activities of three standard antioxidants, quercetin, ascorbic acid, and trolox, were evaluated by Co(II)/ethylenediamine-tetraacetic acid (EDTA)-induced luminol chemiluminescence and the 2,2-diphenyl-1-picrylhydrazyl (DPPH*) free radical assay. Therefore, the aim of this study was to characterise an enzyme-free and time-independent chemiluminescence method for the assessment of the scavenging profile of compounds in a cell-free system using the Co(II)/EDTA-luminol-peroxide system. These results showed that the three standards were efficient and effective in inhibiting both Co(II)/EDTA-induced luminol chemiluminescence and the free radical DPPH*. For all the data obtained in this work, the scavenging activity for the standards tested decreased in the following order: quercetin > trolox > ascorbic acid. The present study has applied a simple and precise procedure for the study of hydroxyl radical scavenging activity by Co(II)/EDTA-induced luminol chemiluminescence, and this was assessed by DPPH* free radical scavenging. PMID- 11395329 TI - Development of a real-time RT-PCR assay for plasma membrane calcium ATPase isoform 1 (PMCA1) mRNA levels in a human breast epithelial cell line. AB - The plasma membrane Ca(2+) pump is a key regulator of cytosolic free Ca(2+). Recent studies have demonstrated the dynamic expression of the plasma membrane Ca(2+) pump in a variety of cell types. Furthermore, alterations in plasma membrane calcium pump activity have now been implicated in human disease. In this study, the development of a technique to quantitatively assess mRNA expression of the human plasma membrane Ca(2+) ATPase (PMCA1) isoform of the plasma membrane Ca(2+) pump, using a real-time reverse transcriptase-polymerase chain reaction (real-time RT-PCR) assay in a human breast epithelial cell line (MCF-7) is described. The sequences of the PMCA1 primers and probe for real-time RT-PCR are presented. The results also indicate that PMCA1 mRNA can be normalized to both 18S ribosomal RNA (18S rRNA) and human glyceraldehyde-3-phosphate dehydrogenase (hGAPDH) in MCF-7 cells. Real-time RT-PCR will be most useful in assessing PMCA1 mRNA expression in cases where only low amounts of RNA are available and/or when numerous samples must be assessed simultaneously. PMID- 11395330 TI - Assessment of in vitro and in vivo recovery of gallamine using microdialysis. AB - The application of microdialysis technique for the investigation of pharmacokinetics and pharmacodynamics of drugs requires careful assessment of probe performance to ensure validity of the data obtained using this technique. The aim of this study was to establish and validate the microdialysis technique for investigation of the pharmacokinetics and pharmacodynamics of the neuromuscular blocker, gallamine. In vitro recovery of gallamine from the microdialysis probe when different perfusion flow rates were employed was evaluated leading to selection of a flow rate of 2 microl/min with 15-min sampling intervals for the subsequent studies. In vitro recovery of gallamine from the microdialysis probe was independent of concentration, stable over an 8-h period and reproducible. Comparable in vitro recoveries were obtained by different established approaches including recovery estimation by gain, loss and the zero-net flux (ZNF) method. Recovery by loss was used to study the in vivo recovery of gallamine from rat muscle tissue. The in vivo recovery was stable over a 5.5-h sampling period. In vitro performance of the probe subsequent to the in vivo study remained stable supporting reusage of the probe. These data highlight the importance of a systematic examination of microdialysis probe validation. PMID- 11395331 TI - Smokeless tobacco extracts modulate exogenous gene expression in early passage cultured human oral epithelial cells: an in vitro system to study chemical and viral enhancer/promoter interaction. AB - The increased risk for cancers of the oral cavity from smokeless tobacco use may reflect the interaction of tobacco with genetic factors, such as oncogenes, and other exogenous factors, such as viruses. An in vitro system was developed based on expression of the chloramphenicol acetyltransferase (CAT) reporter gene to study interactions of chemical treatments with viral enhancer/promoters in early passage cell cultures of oral cavity-derived epithelial cells. Expression of CAT in transfected cells was significantly greater with CAT under the control of the cytomegalovirus immediate early enhancer/promoter (pCEP4/CAT) compared to the Rous sarcoma virus long terminal repeat enhancer/promoter (pRSV-cat) and the simian virus 40 (SV40) early promoter (pSV2-cat). No CAT expression was detected using corresponding control plasmids without the CAT reporter gene. Using this system, smokeless tobacco extracts prepared from either dry snuff or moist snuff delayed maximum CAT expression from Day 4 to Day 5, with sustained, significantly increased CAT expression at 12 days compared to the declining CAT expression observed in untreated control cells. Smokeless tobacco extracts can modulate intracellular gene expression. This system provides an in vitro model to test specificity of toxic agents on enhancer/promoter activity and the interaction on exogenous gene expression. PMID- 11395332 TI - Phenotypic stability of chick cardiomyocytes in serum-free media. Preservation of muscarinic receptor expression. AB - Chick cardiomyocytes cultured in fetal bovine serum (FBS)-supplemented media are phenotypically unstable, becoming noncontractile and unresponsive to stimuli after several days. We report a culturing protocol that preserves the differentiated cardiomyocyte phenotype for at least 9 days in culture. Cardiomyocytes isolated from 11-day chicken embryos, and cultured in either Dulbecco's Modified Earle's Medium (DMEM)/Ham's F12 medium with N-2 supplement or Medium 199 (M199) with 10% FBS continued to beat spontaneously for 4-5 days; only cells cultured in N-2-supplemented medium exhibited spontaneous beating beyond 5 days. Immunostaining for alpha-actinin after 9 days in culture revealed that myofibrils persisted in N-2-supplemented cells, while no myofibrils were observed in the FBS-supplemented cells. For cells in FBS-supplemented media, [3H]thymidine incorporation rates were 7.5 and 3 times greater than that of cells in N-2 supplemented media at Days 4 and 9 in culture, respectively. The effect of growth media on the binding parameters of the muscarinic antagonist, [3H]N-methyl scopolamine (NMS), was also compared. While B(max) decreased 34% between Days 4 and 9 for cells maintained in N-2-supplemented media, a 77% decrease was observed for cells cultured in FBS-supplemented media. The phenotypic stability of this preparation makes it feasible for the first time to use these cells in experiments that require more than 4 days to complete. PMID- 11395333 TI - Comparison of the glucose oxidase method for glucose determination by manual assay and automated analyzer. AB - In experimental models of diabetes, glucose levels in plasma and blood are commonly determined by colorimetric assay and by automated analyzers based on the glucose oxidase conversion of glucose and O2 to gluconate and H2O2. We have compared the glucose levels obtained by these two methods in control Wistar rats, streptozotocin diabetic Wistar rats, Zucker fa/fa fatty rats and Zucker Diabetic Fatty rats. We found that the manual glucose assay and the glucose analyzer produced comparable values up to concentrations of about 25 mM. Above this level, samples should be diluted. PMID- 11395334 TI - Is sticky blood a treatable determinant of cognitive decline and of dementia? PMID- 11395335 TI - Behavioural symptoms in Alzheimer's disease: validity of targets and present treatments. PMID- 11395336 TI - Preventive home visits for older people: defining criteria for success. PMID- 11395337 TI - Breathlessness and quality of life in old age. PMID- 11395338 TI - Stroke education: can we rise to the challenge? PMID- 11395339 TI - Nutritional assessment: a simple step forward. PMID- 11395340 TI - Appraising clinical trial evidence for elderly people: special considerations. PMID- 11395341 TI - Sexuality in the older person. PMID- 11395342 TI - Inadequacies in the provision of information to stroke patients and their families. PMID- 11395343 TI - Cognitive function and blood rheology: results from the Caerphilly cohort of older men. AB - OBJECTIVES: Vascular lesions are important contributors to cerebral disease, yet relatively little work has been done on relationships between haematological factors and cognitive function. We have explored these relationships in the Caerphilly cohort of older men. DESIGN: The development of vascular disease and the decline in cognitive function are being studied in a cohort of older men in Caerphilly, South Wales, UK. We have recorded a range of life-style, dietary, lipid, haematological and rheological factors which are, or may be, predictive of vascular disease. We have examined the relationships between these and both incident myocardial infarction and stroke, and identified factors relevant to cognitive function. SETTING: A community-based study based upon a representative population sample of older men. We collected data for the study from around 90% of the survivors of the original cohort. SUBJECTS: 2154 men who were aged 55-69 years at the time blood was taken for the haematological tests and cognitive function was tested. MAIN OUTCOME MEASURES: We present data from the AH4 test of general cognitive performance and from choice reaction time, a test of vigilance. RESULTS: There is a U-shaped relationship between haematocrit and cognitive function, the best cognitive function scores occurring at a mean haematocrit of 0.46. The relationship is significant for the choice reaction time (P < 0.05). The relationship between haematocrit and the AH4 test score is also curvilinear, but it is not significant. Plasma viscosity showed strong and robust relationships, with significantly better cognitive performance and faster reaction times at lower levels of viscosity (both relationships P < 0.001). Plasma fibrinogen concentration did not show any significant relationships with cognitive function. All these relationships are present, but are much weaker, with rheological tests that had been carried out 5 years before the cognitive testing. CONCLUSIONS: Blood rheology, as estimated by both haematocrit and plasma viscosity, is a significant determinant of cognitive function in older men. On the other hand, the thrombotic potential of blood, as indicated by fibrinogen level, shows no significant relationships. The relationships with rheology seem to be direct, presumably through blood flow at the time of testing, rather than through underlying long-term disease processes. PMID- 11395344 TI - Behavioural management of aggression in dementia: a randomized controlled trial. AB - OBJECTIVE: To evaluate the efficacy of a brief behaviour management training programme for family carers of patients with dementia and aggressive behaviour. DESIGN: A randomized controlled trial of a four-session, community-based behaviour management programme. The primary outcome measure was the Rating Scale for Aggressive Behaviour in the Elderly. PARTICIPANTS: 62 patients with dementia, and their co-resident carers. RESULTS: There were no significant differences in aggression scores between behaviour management and control groups at follow-up. After adjusting for baseline differences in Rating Scale for Aggressive Behaviour in the Elderly scores, there was a trend towards a reduction in aggressive behaviour in the patients in the behaviour management group compared with those in the control group (F = 3.37, P = 0.071). CONCLUSIONS: This study adds to the small evidence base for the effectiveness of behavioural management strategies in dementia. PMID- 11395345 TI - Efficacy of a nurse-led multidimensional preventive programme for older people at risk of functional decline. A randomized controlled trial. AB - OBJECTIVE: To verify the efficacy of a multidimensional preventive programme on functional decline of older people. DESIGN: Randomized controlled trial. SETTING: Community of Sherbrooke City, Quebec, Canada. SUBJECTS: A representative sample of individuals aged over 75 living at home and identified to be at risk of functional decline by postal questionnaire (n = 503). INTERVENTION: Subjects randomized to the study group (n = 250) were assessed at home by a nurse on 12 dimensions (including medication, depressive mood, risk of falls, hearing). A report of the assessment was sent to the general practitioner with recommendations for interventions. A monthly telephone contact was carried out by the nurse for surveillance and to verify if the recommendations had been applied. METHODS: The primary outcome--functional decline--was defined as either death, admission to an institution or increase of > or = 5 points on the disability score of the Functional Autonomy Measurement System (SMAF) scale during the reference year. Secondary outcomes were functional autonomy (on the SMAF), well being (General Well-being Schedule), perceived social support (Social Provisions Scale) and use of health care services. RESULTS: Of the 494 subjects who completed the study, 48 (19.6%) of 245 in the study group and 49 (19.7%) of 249 in the control group had functional decline (relative risk 1.00; 95% confidence interval 0.82-1.23). There were no differences between the groups in all secondary outcomes. CONCLUSIONS: This study confirms the inefficacy of multidimensional programmes for preventing functional decline in the older population. More effort should be devoted to improving the efficacy of specific interventions for conditions causing functional decline. PMID- 11395346 TI - Dyspnoea and quality of life in older people at home. AB - OBJECTIVES: To determine the prevalence of dyspnoea in older people at home, measure its impact on function and quality of life, and identify associated cardio-respiratory diseases. DESIGN: Cross-sectional population-based study. METHODS: We sent a modified Medical Research Council (MRC) dyspnoea questionnaire to identify breathlessness in 1404 randomly selected subjects from general practitioner lists of 5002 subjects aged 70 years and over living at home. We visited a further random sample of 500 of these subjects at home and at a study centre. SETTING: Community-based study in South Wales. MAIN OUTCOME MEASURES: Prevalence of dyspnoea (MRC grades 3-5) and its effect on psychological and functional status, and quality of life as measured by Hospital Anxiety and Depression, Nottingham Extended Activities of Daily Living and SF-36 questionnaires. RESULTS: The prevalence of dyspnoea as defined was 32.3% (95% confidence intervals: 30.3, 34.3). Breathless subjects had poorer functional status than non-breathless subjects. They also had poorer physical and mental health and were more likely to be anxious and depressed. The prevalence of left ventricular systolic dysfunction, reversible airways disease and obesity were all higher in those with dyspnoea. CONCLUSIONS: Dyspnoea is common in older people. Given its profound adverse effect on people's lives, dyspnoea is an important public health issue. PMID- 11395347 TI - Validation of the Resident Assessment Instrument triggers in the detection of under-nutrition. AB - OBJECTIVE: To test the validity of Resident Assessment Instrument triggers for people in Danish nursing homes and subjects receiving home care by exploring their ability to identify older people with insufficient energy and protein intake. DESIGN: Regression analysis of insufficient energy or protein intake on each of the Resident Assessment Instrument triggers alone or in combination. SUBJECTS: 38 people receiving home care and 41 living in nursing homes. METHODS: A 4 day estimated dietary record, assessment of energy and protein intake, calculation of body mass index. RESULTS: Food remaining on the plate, weight loss and any trigger present significantly influenced the energy or protein intake of the nursinghome subjects. No triggers reflected the energy or protein intake of the subjects at home. CONCLUSION: Observation of food intake seems to be the best way to detect which older Danish people living in nursing homes or receiving home care have insufficient energy or protein intake. PMID- 11395348 TI - Giant cell arteritis presenting with arm claudication. AB - CASE REPORT: A 72-year-old woman presented after a series of falls with a history of malaise, lethargy, cold hands and arm weakness for several months. She had undergone extensive investigation for what remained an unexplained anaemia during this period. Examination revealed cold hands with absent radial pulses. Axillary Doppler studies confirmed grossly diminished systolic pressures. The erythrocyte sedimentation rate was markedly elevated. An arch aortogram showed obliterative changes selectively affecting the subclavian and axillary arteries. In spite of clinically normal temporal arteries, and the absence of headache, temporal artery biopsy revealed giant cell arteritis. DISCUSSION: Aortic arch syndrome is an important but under-appreciated complication of giant cell arteritis and may be the presenting feature. In this case, it resulted in falls consequent upon arm claudication when using a walking frame. PMID- 11395349 TI - Patient and carer perception of the management of Parkinson's disease after surgery. PMID- 11395350 TI - Markedly increased levels of IL-6 and CA125 in pleural fluid of an elderly person with overlap syndrome of systemic sclerosis and systemic lupus erythematosus. PMID- 11395351 TI - A case of chronic subdural haematoma presenting as a translent ischaemic attack. PMID- 11395352 TI - Hospital-at-home and community care: are they the same? PMID- 11395353 TI - Vitamin D deficiency. PMID- 11395354 TI - Nonconvulsive status epilepticus causing acute confusion. PMID- 11395355 TI - Adherence to recommendations of community-based comprehensive geriatric assessment programmes. PMID- 11395356 TI - Spontaneous rupture of oesophagus (Boerhaave's syndrome) related to rivastigmine. PMID- 11395357 TI - Management of blood pressure after acute stroke: a European perspective. PMID- 11395358 TI - Vertebral osteomyelitis. PMID- 11395359 TI - Parkinson's Disease and Parkinsonism in the Elderly. PMID- 11395360 TI - Endocrinology of Aging. PMID- 11395362 TI - Pten, a protean tumor suppressor. PMID- 11395363 TI - Mechanism of apoptotic regulation of follicular regression: toward understanding the molecular basis for tissue remodeling and regeneration. PMID- 11395364 TI - Putative tumor suppressor loci at 6q22 and 6q23-q24 are involved in the malignant progression of sporadic endocrine pancreatic tumors. AB - Our previous comparative genomic hybridization study on sporadic endocrine pancreatic tumors (EPTs) revealed frequent losses on chromosomes 11q, 3p, and 6q. The aim of this study was to evaluate the importance of 6q losses in the oncogenesis of sporadic EPTs and to narrow down the smallest regions of allelic deletion. A multimodal approach combining polymerase chain reaction-based allelotyping, double-target fluorescence in situ hybridization, and comparative genomic hybridization was used in a collection of 109 sporadic EPTs from 93 patients. Nine polymorphic microsatellite markers (6q13 to 6q25-q27) were investigated, demonstrating a loss of heterozygosity (LOH) in 62.2% of the patients. A LOH was significantly more common in tumors >2 cm in diameter than below this threshold as well as in malignant than in benign tumors. We were able to narrow down the smallest regions of allelic deletion at 6q22.1 (D6S262) and 6q23-q24 (D6S310-UTRN) with LOH-frequencies of 50.0% and 41.2 to 56.3%, respectively. Several promising tumor suppressor candidates are located in these regions. Additional fluorescence in situ hybridization analysis on 46 EPTs using three locus-specific probes (6q21, 6q22, and 6q27) as well as a centromere 6 specific probe revealed complete loss of chromosome 6 especially in metastatic disease. We conclude that the two hot spots found on 6q may harbor putative tumor suppressor genes involved not only in the oncogenesis but maybe also in the malignant and metastatic progression of sporadic EPTs. PMID- 11395365 TI - p53 Involvement in the control of murine hair follicle regression. AB - p53 is a transcription factor mediating a variety of biological responses including apoptotic cell death. p53 was recently shown to control apoptosis in the hair follicle induced by ionizing radiation and chemotherapy, but its role in the apoptosis-driven physiological hair follicle regression (catagen) remains to be elucidated. Here, we show that p53 protein is strongly expressed and co localized with apoptotic markers in the regressing hair follicle compartments during catagen. In contrast to wild-type mice, p53 knockout mice show significant retardation of catagen accompanied by significant decrease in the number of apoptotic cells in the hair matrix. Furthermore, p53 null hair follicles are characterized by alterations in the expression of markers that are encoded by p53 target genes and are implicated in the control of catagen (Bax, Bcl-2, insulin like growth factor binding protein-3). These data suggest that p53 is involved in the control of apoptosis in the hair follicle during physiological regression and imply that p53 antagonists may be useful for the management of hair growth disorders characterized by premature entry into catagen, such as androgenetic alopecia, alopecia areata, and telogen effluvium. PMID- 11395366 TI - Tumorigenic potential of extracellular matrix metalloproteinase inducer. AB - Extracellular matrix metalloproteinase inducer (EMMPRIN), a glycoprotein present on the cancer cell plasma membrane, enhances fibroblast synthesis of matrix metalloproteinases (MMPs). The demonstration that peritumoral fibroblasts synthesize most of the MMPs in human tumors rather than the cancer cells themselves has ignited interest in the role of EMMPRIN in tumor dissemination. In this report we have demonstrated a role for EMMPRIN in cancer progression. Human MDA-MB-436 breast cancer cells, which are tumorigenic but slow growing in vivo, were transfected with EMMPRIN cDNA and injected orthotopically into mammary tissue of female NCr nu/nu mice. Green fluorescent protein was used to visualize metastases. In three experiments, breast cancer cell clones transfected with EMMPRIN cDNA were considerably more tumorigenic and invasive than plasmid transfected cancer cells. Increased gelatinase A and gelatinase B expression (demonstrated by in situ hybridization and gelatin substrate zymography) was demonstrated in EMMPRIN-enhanced tumors. In contrast to de novo breast cancers in humans, human tumors transplanted into mice elicited minimal stromal or inflammatory cell reactions. Based on these experimental studies and our previous demonstration that EMMPRIN is prominently displayed in human cancer tissue, we propose that EMMPRIN plays an important role in cancer progression by increasing synthesis of MMPs. PMID- 11395368 TI - Differential loss of chromosome 11q in familial and sporadic parasympathetic paragangliomas detected by comparative genomic hybridization. AB - Parasympathetic paragangliomas (PGLs) represent neuroendocrine tumors arising from chief cells in branchiomeric and intravagal paraganglia, which share several histological features with their sympathetic counterpart sympathoadrenal paragangliomas. In recent years, genetic analyses of the familial form of PGL have attracted considerable interest. However, the majority of paragangliomas occurs sporadically and it remains to be determined whether the pathogenesis of sporadic paraganglioma resembles that of the familial form. Furthermore, data on comparative genetic aberrations are scarce. To provide fundamental cytogenetic data on sporadic and hereditary PGLs, we performed comparative genomic hybridization using directly fluorochrome-conjugated DNA extracted from 12 frozen and 4 paraffin-embedded tumors. The comparative genomic hybridization data were extended by loss of heterozygosity analysis of chromosome 11q. DNA copy number changes were found in 10 (63%) of 16 tumors. The most frequent chromosomal imbalance involved loss of chromosome 11. Six of seven familial tumors and two of nine sporadic tumors showed loss of 11q (86% versus 22%, P = 0.012). Deletions of 11p and 5p were found in two of nine sporadic tumors. We conclude that overall DNA copy number changes are infrequent in PGLs compared to sympathetic paragangliomas and that loss of chromosome 11 may be an important event in their tumorigenesis, particularly in familial paragangliomas. PMID- 11395369 TI - Vascular smooth muscle cells of recipient origin mediate intimal expansion after aortic allotransplantation in mice. AB - Intimal expansion by vascular smooth muscle cells (SMCs) is a characteristic feature of graft vascular disease. Whether graft intimal SMCs arise from donor or recipient tissue is not well established but has important pathogenetic implications. We examined for the presence of male cells in the expanded intima of sex-mismatched mouse aortic allografts (C57BL/6-to-BALB/c) at 30 or 60 days after transplant by in situ hybridization using a Y-chromosome probe. Study groups included male-to-female allografts, female-to-male allografts, and female to-female allografts in recipients previously engrafted with male bone marrow. Although intimal expansion developed in all allografts, male-to-female allografts lacked Y-chromosome-positive intimal cells. In contrast, such cells were abundant in female-to-male allografts and most of these cells co-labeled for smooth muscle alpha-actin by immunostain. Female-to-female allografts in recipients with male bone marrow showed a limited number of intimal Y-chromosome-positive cells. However, none of these clearly co-labeled for smooth muscle alpha-actin and their numbers declined throughout time, consistent with graft-infiltrating inflammatory cells. We conclude that intimal expansion of mouse aortic allografts is mediated by SMCs that originated from the recipient. There was little evidence of their derivation from the bone marrow, suggesting instead the adjacent host aorta as the primary source of intimal SMCs. PMID- 11395367 TI - Adult-derived stem cells from the liver become myocytes in the heart in vivo. AB - Recent evidence suggests that adult-derived stem cells, like their embryonic counterparts, are pluripotent. These simple, undifferentiated and uncommitted cells are able to respond to signals from their host tissue microenvironment and differentiate, producing progeny that display a phenotype characteristic of the mature cells of that tissue. We used a clonal stem cell line (termed WB-F344) that was derived from an adult male rat liver to investigate the possibility that uncommitted stem cells from a nonmyogenic tissue source would respond to the tissue microenvironment of the heart in vivo and differentiate into cardiac myocytes. Male WB-F344 cells that carry the Escherichia coli beta-galactosidase gene were identified in the left ventricular myocardium of adult female nude mice 6 weeks after transplantation. We confirmed the presence of a rat Y-chromosome specific repetitive DNA sequence exclusively in the beta-galactosidase-positive myocytes by polymerase chain reaction and fluorescence in situ hybridization. Immunohistochemistry, using a cardiac troponin T-specific monoclonal antibody, and ultrastructural analysis confirmed a cardiac myocyte phenotype of the stem cell-derived myocytes. The beta-galactosidase-positive myocytes ranged from < 20 microm to 110 microm in length. The longer of these cells contained well organized sarcomeres and myofibrils, and formed intercalated disks and gap junctions with endogenous (host-derived) myocytes, suggesting that WB-F344 derived myocytes participate in the function of the cardiac syncytium. These results demonstrate that adult liver-derived stem cells respond to the tissue microenvironment of the adult heart in vivo and differentiate into mature cardiac myocytes. PMID- 11395370 TI - Regulated on activation, normal T-cell-expressed and -secreted mRNA expression in normal endometrium and endometriotic implants: assessment of autocrine/paracrine regulation by in situ hybridization. AB - Chemoattraction of macrophages and T cells into the normal endometrium and inflammatory sites within endometriotic foci is mediated by chemokine gene expression. mRNA transcripts encoding regulated on activation, normal T-cell expressed and -secreted (RANTES), a monocyte and T-cell chemokine, were demonstrated in the stroma of normal endometrium and endometriotic implants using in situ mRNA hybridization. Epithelial glands failed to express RANTES mRNA. In histological serial sections, we observed CD68-positive macrophages in the stroma of endometriotic implants adjacent to regions with prominent RANTES mRNA hybridization. In adjacent sections, monoclonal antibodies against tumor necrosis factor (TNF)-alpha showed this cytokine to be localized to stromal and epithelial compartments of the endometriotic implant with weak staining in unaffected ovarian tissue. Subconfluent monolayers of endometriotic stromal cells were tested for RANTES gene expression in situ, but we could only detect RANTES mRNA in isolated stromal cells after treatment with TNF-alpha. No RANTES mRNA was observed in unstimulated stromal cells or TNF-alpha stimulated or unstimulated epithelial cells. The data are consistent with a model in which proinflammatory cytokines (eg, TNF-alpha) induce RANTES gene expression limited to specific cells within endometrial and endometriotic stroma. Production of this chemokine, in turn, stimulates recruitment of CD68-positive macrophages into these tissues. PMID- 11395371 TI - Frequent FGFR3 mutations in papillary non-invasive bladder (pTa) tumors. AB - We recently identified activating mutations of fibroblast growth factor receptor 3 (FGFR3) in bladder carcinoma. In this study we assessed the incidence of FGFR3 mutations in a series of 132 bladder carcinomas: 20 carcinoma in situ (CIS), 50 pTa, 19 pT1, and 43 pT2-4. All 48 mutations identified were identical to the germinal activating mutations that cause thanatophoric dysplasia, a lethal form of dwarfism. The S249C mutation, found in 33 of the 48 mutated tumors, was the most common. The frequency of mutations was higher in pTa tumors (37 of 50, 74%) than in CIS (0 of 20, 0%; P < 0.0001), pT1 (4 of 19, 21%; P < 0.0001) and pT2-4 tumors (7 of 43, 16%; P < 0.0001). FGFR3 mutations were detected in 27 of 32 (84%) G1, 16 of 29 (55%) G2, and 5 of 71 (7%) G3 tumors. This association between FGFR3 mutations and low grade was highly significant (P < 0.0001). FGFR3 is the first gene found to be mutated at a high frequency in pTa tumors. The absence of FGFR3 mutations in CIS and the low frequency of FGFR3 mutations in pT1 and pT2-4 tumors are consistent with the model of bladder tumor progression in which the most common precursor of pT1 and pT2-4 tumors is CIS. PMID- 11395372 TI - Molecular cytogenetic evaluation of gastric cardia adenocarcinoma and precursor lesions. AB - Analyses of cancer incidence data in the United States and Western Europe revealed steadily rising rates over the past decades of adenocarcinomas of the esophagus and gastric cardia. Genetic information on gastric cardia adenocarcinoma and its preneoplasias is sparse. We have used comparative genomic hybridization to obtain a genome-wide overview of 20 archival gastric cardia adenocarcinomas and 10 adjacent preneoplastic lesions (4 metaplasias, 1 low-grade dysplasia, 5 high-grade dysplasias). Multiple genetic alterations were discriminated in all adenocarcinomas. Frequent loss (> or =25% of all tumors) was detected, in decreasing order of frequency, on 5q, 18q, 4q, 3p, 9p, 2q, 11q, 14q, 21q, 4p, 9q, 16q, 1p, and 8p. Frequent gain (> or =25% of all tumors) was disclosed, in decreasing order of frequency, on 20q, 7p, 8q, 1q, 7q, 20p, 17q, 13q, Xp, 6q, 8p, 19q, 5p, 6p, and Xq. Loss of the Y chromosome was found in 60% of male cases. High level amplification was frequently (>10% of all tumors) detected on 7q21, 8p22, 12p11.2, 17q12-q21, and 19q13.1-q13.2. The precursor lesions showed multiple aberrations in all high-grade dysplasias, whereas few genetic changes were discerned in LGD and metaplasias. High level amplifications were also found in high-grade dysplasias, ie, on 7q21, 8p22, and 17q12-q21. Moreover, the percentage of aberrations was not significantly different for invasive carcinomas or high-grade dysplasias. Approximately 70% of the precursor aberrations were also present in the adjacent carcinoma. Minimal overlapping regions in the preneoplasias included loss on 18q12-q21 and gains on 8q23 and 17q12-q21, suggesting involvement of genes residing in these regions. In conclusion, we have (i) created a map of genetic alterations in gastric cardia adenocarcinomas and (ii) provided evidence for the presence of a metaplasia dysplasia-carcinoma sequence in this poorly understood type of cancer. PMID- 11395373 TI - Involvement of the receptor for formylated peptides in the in vivo anti-migratory actions of annexin 1 and its mimetics. AB - An innovative avenue for anti-inflammatory therapy is inhibition of neutrophil extravasation by potentiating the action of endogenous anti-inflammatory mediators. The glucocorticoid-inducible protein annexin 1 and derived peptides are effective in inhibiting neutrophil extravasation. Here we tested the hypothesis that an interaction with the receptor for formylated peptide (FPR), so far reported only in vitro, could be the mechanism for this in vivo action. In a model of mouse peritonitis, FPR antagonists abrogated the anti-migratory effects of peptides Ac2-26 and Ac2-12, with a partial reduction in annexin 1 effects. A similar result was obtained in FPR (knock-out) KO mice. Binding of annexin 1 to circulating leukocytes was reduced (>50%) in FPR KO mice. In vitro, annexin binding to peritoneal macrophages was also markedly reduced in FPR KO mice. Finally, evidence of direct annexin 1 binding to murine FPR was obtained with HEK 293 cells transfected with the receptor. Overall, these results indicate a functional role for FPR in the anti-migratory effect of annexin 1 and derived peptides. PMID- 11395374 TI - In situ characterization of genetically targeted (green fluorescent) single cells and their microenvironment in an adoptive host. AB - Stable expression of transgene-encoded enhanced green fluorescence protein (eGFP) was used as a sensitive and specific marker to detect in situ donor cells engrafted into different tissues of adoptive hosts. eGFP(+) lymphoid or myeloid cells (eg, CD4(+) T cells or bone marrow-derived dendritic cells) from eGFP transgenic C57BL/6 donor mice were injected into congenic, immunodeficient RAG1( /-) C57/BL6 hosts. eGFP(+) cells were detected in the adoptive host from 2 days to 4 weeks after transfer using an optimized method of fixed cryopreservation to process the tissue. This allowed the simple, sensitive, and specific detection of eGFP(+) donor cells in histological sections of transplanted hosts. We further demonstrate that this technique can be combined with other established labeling methods such as 1) immunofluorescent labeling to characterize the host cells interacting with engrafted cells and to determine the phenotype of the engrafted cells in situ; 2) terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling staining to detect apoptotic death of engrafted and autochthonous cell populations; and 3) fluorescent antibody labeling of incorporated bromodeoxyuridine to measure the fraction of proliferating cells in the graft. PMID- 11395375 TI - A new method for histological microdissection utilizing an ultrasonically oscillating needle: demonstrated by differential mRNA expression in human lung carcinoma tissue. AB - Molecular analysis of microdissected tissue samples is used for analyzing tissue heterogeneity of histological specimens. We have developed a rapid one-step microdissection technique, which was applied for the selective procurement of tissue areas down to a minimum of 10 cell profiles. The special features of our microdissection system consist of an ultrasonically oscillating needle and a piezo-driven micropipette. The validity of this technique is demonstrated in human lung large-cell carcinoma by real-time quantitative reverse transcriptase polymerase chain reaction assays of vimentin, cyclin D1, and carcinoembryonic antigen after linear RNA amplification. mRNA expression values of microdissected samples scattered around those of bulk tumor tissue and showed differential mRNA expression between samples of tumor parenchyma and supportive stromal cells for vimentin and carcinoembryonic antigen as confirmed by immunohistochemistry. In conclusion, this procedure requires simple equipment, is easily performed, and delivers microdissected tissue samples of oligocellular clusters suitable for further molecular analysis. PMID- 11395376 TI - Immunohistochemical detection of cell growth fraction in formalin-fixed and paraffin-embedded murine tissue. AB - Monoclonal antibody MIB-1 is a reliable tool for determining proliferating cells in human tissues, but does not react with the homologous mouse antigen and is therefore useless in experimental pathology using mice as model systems. Standard method for assessment of cellular proliferation in formalin-fixed, paraffin embedded murine tissues is immunohistochemical detection of DNA synthesis using antibodies against exogenously injected 5-bromodeoxyuridine (BrdU), which is a tedious procedure and not useful for routine investigations. We tested monoclonal antibody MIB-5 and monoclonal and polyclonal anti-MCM3 antibodies as immunohistochemical proliferation markers for paraffin-embedded nonneoplastic and neoplastic tissues of wild-type and transgenic mice, compared to anti-BrdU immunostaining. Percentage of proliferating cells was determined with continuously decreasing antibody dilutions. Percentages of MIB-5 and anti-BrdU immunostained cells correlated strongly, as well as percentage of MIB-5-decorated cells and frequency of mitotic figures. Anti-MCM3 antibodies labeled significantly higher percentages of cells than anti-BrdU or MIB-5, and showed a linear decrease with increasing antibody dilutions. We conclude that MIB-5 detects reliably the cell growth fraction in formalin fixed, paraffin-embedded murine tissues, bypassing methodological drawbacks of BrdU. Anti-MCM3 antibodies are less useful for determination of proliferating cells although they might detect the fraction of cells remaining competent for proliferation. PMID- 11395377 TI - A precise and efficient stereological method for determining murine lung metastasis volumes. AB - We have developed a computer-assisted stereological method based on unbiased principles for estimating metastasis volumes in mouse lungs. We evaluated this method using the transplantable Lewis lung carcinoma. Twenty-one days after subcutaneous inoculation of 10(6) Lewis lung cells into C57BL/6J mice, the mice had primary tumors with an average volume of 2300 mm(3). After perfusion fixation, the lungs were removed, embedded in OCT compound, snap-frozen, and processed for stereology. The metastasis volumes were estimated by application of the Cavalieri principle after evaluation of single sections from several evenly distributed tissue levels. The metastasis volume in a group of nine mice varied between 0.01 and 14.4 mm(3), with an average of 6.1 mm(3). The coefficient of variation was 0.9. The coefficient of error of the volume estimation was determined in five cases and varied from 0.08 to 0.23. Thus, the variation on the metastasis volumes that is achieved by this method contributes very little, 2.5%, to the total variance within the group of mice. In conclusion, we have developed an efficient and unbiased method to determine the metastasis burden in mouse lungs. PMID- 11395378 TI - Expression profiling of ductal carcinoma in situ by laser capture microdissection and high-density oligonucleotide arrays. AB - Gene expression profiling through the use of nucleic acid arrays is a powerful method for the molecular classification of human neoplasms. Laser capture microdissection is an equally useful technique to selectively isolate defined cell populations from heterogeneous histological tissue sections. In this report, we demonstrate how a modest use of laser capture microdissection is sufficient to isolate nanogram quantities of high-quality RNA. Together with the use of several internal standards and microcapillary electrophoresis of input RNA, two rounds of linear molecular amplification have been used to generate sufficient quantities of labeled target for hybridization to high-density oligonucleotide expression arrays. Results demonstrate that the technique is reproducible, generates only modest biasing of the original transcript population, and is comparable to the sensitivity achieved with standard methodology. Using this approach, we have compared the expression profiles of nonmalignant human breast epithelium and adjacent ductal carcinoma in situ lesions from breast cancer patients. Several genes, previously implicated in human breast cancer progression, demonstrate differential expression among the microdissected cell populations. PMID- 11395379 TI - Potential role of modifier genes influencing transforming growth factor-beta1 levels in the development of vascular defects in endoglin heterozygous mice with hereditary hemorrhagic telangiectasia. AB - Hereditary hemorrhagic telangiectasia (HHT) is an autosomal dominant disorder because of mutations in the genes coding for endoglin (HHT1) or ALK-1 (HHT2). The disease is associated with haploinsufficiency and a murine model was obtained by engineering mice that express a single Endoglin allele. Of a total of 171 mice that were observed for 1 year, 50 developed clinical signs of HHT. Disease prevalence was high in 129/Ola strain (72%), intermediate in the intercrosses (36%), and low in C57BL/6 backcrosses (7%). Most mice first presented with an ear telangiectasia and/or recurrent external hemorrhage. One-third of mice with HHT showed severe vascular abnormalities such as dilated vessels, hemorrhages, liver and lung congestion, and/or brain and heart ischemia. Disease sequelae included stroke, hydrocephalus, fatal hemorrhage, and congestive heart failure. Thus the murine model reproduces the multiorgan manifestations of the human disease. Levels of circulating latent transforming growth factor (TGF)-beta1 were significantly lower in the 129/Ola than in the C57BL/6 strain. Intercrosses and 129/Ola mice expressing reduced endoglin also showed lower plasma TGF-beta1 levels than control. These data suggest that modifier genes involved in the regulation of TGF-beta1 expression act in combination with a single functional copy of endoglin in the development of HHT. PMID- 11395380 TI - Dimerization co-factor of hepatocyte nuclear factor 1/pterin-4alpha-carbinolamine dehydratase is necessary for pigmentation in Xenopus and overexpressed in primary human melanoma lesions. AB - Dimerization co-factor of hepatocyte nuclear factor 1 (HNF1)/pterin-4alpha carbinolamine dehydratase (DCoH/PCD) is both a positive co-factor of the HNF1 homeobox transcription factors and thus involved in gene regulation as well as an enzyme catalyzing the regeneration of tetrahydrobiopterin. Dysfunction of DCoH/PCD is associated with the human disorders hyperphenylalaninemia and vitiligo. In Xenopus, overexpression of the protein during development induces ectopic pigmentation. In this study loss of function experiments using DCoH/PCD specific antibodies demonstrated that the protein is also absolutely necessary for pigment cell formation in Xenopus. In normal human skin DCoH/PCD protein is weakly expressed in the basal layer of the epidermis that consists of keratinocytes and melanocytes. Whereas only 4 of 25 benign nevi reacted with DCoH/PCD-specific antibodies, high protein levels were detectable in melanoma cell lines and 13 of 15 primary malignant melanoma lesions. The comparison with the commonly used melanoma markers S100 and HMB45 demonstrated that DCoH/PCD has an overlapping but distinct expression pattern in melanoma lesions. In addition to human colon cancer, this is the second report about the overexpression of DCoH/PCD in human tumor cells indicating that the protein might be involved in cancerogenesis. PMID- 11395381 TI - Differential expression of protease-activated receptors-1 and -2 in stromal fibroblasts of normal, benign, and malignant human tissues. AB - The serine proteases thrombin and trypsin are among many factors that malignant cells secrete into the extracellular space to mediate metastatic processes such as cellular invasion, extracellular matrix degradation, angiogenesis, and tissue remodeling. The degree of protease secretion from malignant cells has been correlated to their metastatic potential. Protease activated receptors (PAR)-1 and -2, which are activated by thrombin and trypsin respectively, have not been extensively characterized in human tumors in situ. We investigated the presence of PAR-1 and PAR-2 in human normal, benign and malignant tissues using immunohistochemistry and in situ hybridization. Our results demonstrate PAR-1 and PAR-2 expression in the tumor cells, mast cells, macrophages, endothelial cells, and vascular smooth muscle cells of the metastatic tumor microenvironment. Most notably, an up-regulation of PAR-1 and PAR-2 observed in proliferating, smooth muscle actin (SMA)-positive stromal fibroblasts surrounding the carcinoma cells was not observed in normal or benign conditions. Furthermore, in vitro studies using proliferating, SMA-positive, human dermal fibroblasts, and scrape-wounded human dermal fibroblasts demonstrated the presence of PAR-1 and PAR-2 not detected in quiescent, SMA-negative cultures. PAR-1 and PAR-2 in the cells forming the tumor microenvironment suggest that these receptors mediate the signaling of secreted thrombin and trypsin in the processes of cellular metastasis. PMID- 11395382 TI - Time course of endothelial cell proliferation and microvascular remodeling in chronic inflammation. AB - Angiogenesis and vascular remodeling are features of many chronic inflammatory diseases. When diseases evolve slowly, the accompanying changes in the microvasculature would seem to be similarly gradual. Here we report that the rate of endothelial cell proliferation and the size of blood vessels increases rapidly after the onset of an infection that leads to chronic inflammatory airway disease. In C3H mice inoculated with Mycoplasma pulmonis, the tracheal microvasculature, made visible by perfusion of Lycopersicon esculentum lectin, rapidly enlarged from 4 to 7 days after infection and then plateaued. Diameters of arterioles, capillaries, and venules increased on average 148, 214, and 74%, respectively. Endothelial cell proliferation, measured by bromodeoxyuridine (BrdU) labeling, peaked at 5 days (18 times the pathogen-free value), declined sharply until day 9, but remained at approximately 3 times the pathogen-free value for at least 28 days. Remodeled capillaries and venules were sites of focal plasma leakage and extensive leukocyte adherence. Most systemic manifestations of the infection occurred well after the peak of endothelial proliferation, and the humoral immune response to M. pulmonis was among the latest, increasing after 14 days. These data show that endothelial cell proliferation and microvascular remodeling occur at an early stage of chronic airway disease and suggest that the vascular changes precede widespread tissue remodeling. PMID- 11395383 TI - Expression of inducible nitric oxide synthase and nitrotyrosine in multiple sclerosis lesions. AB - Nitric oxide generated by the inducible form of nitric oxide synthase (iNOS) may contribute to the pathogenesis of multiple sclerosis (MS). In this report, we studied postmortem tissues of MS patients for the expression of iNOS by in situ hybridization and immunocytochemistry. Immunocytochemistry for nitrotyrosine, a putative footprint for peroxynitrite formation was also performed. In acute MS lesions, intense reactivity for iNOS mRNA and protein was detected in reactive astrocytes throughout the lesion and in adjacent normal appearing white matter. Staining of macrophages, inflammatory cell infiltrates, and endothelial cells was variable from case to case, but generally detected only in acute lesions. In chronic MS lesions reactive astrocytes at the lesion edge were positive for iNOS whereas the lesion center was nonreactive. Normal appearing white matter demonstrated little reactivity, as did tissues from noninflamed control brains. Staining for nitrotyrosine was also detected in acute but not chronic MS lesions, and displayed a diffuse parenchymal, membranous, and perivascular pattern of immunoreactivity. These results support the conclusion that iNOS is induced in multiple cell types in MS lesions and that astrocyte-derived nitric oxide could be important in orchestrating inflammatory responses in MS, particularly at the blood-brain barrier. PMID- 11395384 TI - p53 cellular localization and function in neuroblastoma: evidence for defective G(1) arrest despite WAF1 induction in MYCN-amplified cells. AB - This study investigated the hypothesis that p53 accumulation in neuroblastoma, in the absence of mutation, is associated with functional inactivation, which interferes with downstream mediators of p53 function. To test this hypothesis, p53 expression, location, and functional integrity was examined in neuroblastoma by irradiating 6 neuroblastoma cell lines and studying the effects on p53 transcriptional function, cell cycle arrest, and induction of apoptosis, together with the transcriptional function of p53 after irradiation in three ex vivo primary, untreated neuroblastoma tumors. p53 sequencing showed five neuroblastoma cell lines, two of which were MYCN-amplified, and that all of the tumors were wild-type for p53. p53 was found to be predominantly nuclear before and after irradiation and to up-regulate the p53 responsive genes WAF1 and MDM2 in wild type p53 cell lines and a poorly-differentiated neuroblastoma, but not a differentiating neuroblastoma or the ganglioneuroblastoma part of a nodular ganglioneuroblastoma in short term culture. This suggests intact p53 transcriptional activity in proliferating neuroblastoma. Irradiation of wild-type p53 neuroblastoma cell lines led to G(1) cell cycle arrest in cell lines without MYCN amplification, but not in those with MYCN amplification, despite induction of WAF1. This suggests MYCN amplification may alter downstream mediators of p53 function in neuroblastoma. PMID- 11395385 TI - Quantitative assessment of the rat intrahepatic biliary system by three dimensional reconstruction. AB - The anatomical details of the biliary tree architecture of normal rats and rats in whom selective proliferation was induced by feeding alpha naphthylisothiocyanate (ANIT) were reconstructed in three dimension using a microscopic-computed tomography scanner. The intrahepatic biliary tree was filled with a silicone polymer through the common bile duct and each liver lobe embedded in Bioplastic; specimens were then scanned by a microscopic-computed tomography scanner and modified Feldkamp cone beam backprojection algorithm applied to generate three-dimensional images. Quantitative analysis of bile duct geometry was performed using a customized software program. The diameter of the bile duct segments of normal and ANIT-fed rats progressively decreased with increasing length of the biliary tree. Diameter of bile ducts from ANIT-fed rats (range, 21 to 264 microm) was similar to that of normal rats (22 to 279 microm). In contrast, the number of bile duct segments along the major branch reproducibly doubled, the length of the bile duct segments decreased twofold, and the length of the biliary tree remained unchanged after ANIT feeding. Moreover, the total volume of the biliary tree of ANIT-fed rats was significantly greater (855 microl) than in normal rats (47 microl). Compared with normal rats, the total surface area of the biliary tree increased 26 times after ANIT-induced bile duct proliferation. Taken together, these observations quantitate the anatomical remodeling after selective cholangiocyte proliferation and strongly suggest that the proliferative process involves sprouting of new side branches. Our results may be relevant to the mechanisms by which ducts proliferate in response to hepatic injury and to the hypercholeresis that occurs after experimentally induced bile duct proliferation. PMID- 11395386 TI - A distinctive pediatric renal neoplasm characterized by epithelioid morphology, basement membrane production, focal HMB45 immunoreactivity, and t(6;11)(p21.1;q12) chromosome translocation. AB - We report two cases of a hitherto undescribed pediatric renal neoplasm that is distinctive at the morphological, immunohistochemical, ultrastructural, and cytogenetic levels. On light microscopy, the tumors are composed of nests of polygonal, clear to eosinophilic cells associated with a subpopulation of smaller cells that surround hyaline material. Despite their epithelioid morphology, these tumors do not label immunohistochemically for epithelial markers but instead label focally for melanocytic markers HMB45 and Melan A. The hyaline material is positive with periodic acid-Schiff and methenamine-silver histochemical stains, and labels immunohistochemically for type 4 collagen. Ultrastructural examination confirms that it represents basement membrane material. Cytogenetic analysis reveals the identical t(6;11)(p21.1;q12) chromosome translocation as the sole abnormality in these two tumors, confirming their identity and distinctive nature. PMID- 11395388 TI - Expression of microtubule-associated protein 2 in benign and malignant melanocytes: implications for differentiation and progression of cutaneous melanoma. AB - Cutaneous melanocytic neoplasms are known to acquire variable characteristics of neural crest differentiation. Melanocytic nevus cells in the dermis and desmoplastic melanomas often display characteristics of nerve sheath differentiation. The extent and nature of neuronal differentiation characteristics displayed by primary and metastatic melanoma cells are not well understood. Here, we describe induction of a juvenile isoform of microtubule associated protein 2 (MAP-2c) in cultured metastatic melanoma cells by the differentiation inducer hexamethylene bisacetamide. Up-regulation of this MAP-2 isoform, a marker for immature neurons, is accompanied by extended dendritic morphology and down-regulation of tyrosinase-related protein 1 (TYRP1/gp75), a melanocyte differentiation marker. In a panel of cell lines that represent melanoma tumor progression, MAP-2c mRNA and the corresponding approximately 70-kd protein could be detected predominantly in primary melanomas. Immunohistochemical analysis of 61 benign and malignant melanocytic lesions showed abundant expression of MAP-2 protein in melanocytic nevi and in the in situ and invasive components of primary melanoma, but only focal heterogeneous expression in a few metastatic melanomas. In contrast, MAP-2-positive dermal nevus cells and the invasive cells of primary melanomas were TYRP1-negative. This reciprocal staining pattern in vivo is similar to the in vitro observation that induction of the neuronal marker MAP-2 in metastatic melanoma cells is accompanied by selective extinction of the melanocytic marker TYRP1. Our data show that neoplastic melanocytes, particularly at early stages, retain the plasticity to express the neuron-specific marker MAP-2. These observations are consistent with the premise that both benign and malignant melanocytes in the dermis can express markers of neuronal differentiation. PMID- 11395387 TI - Frequent loss of PTEN expression is linked to elevated phosphorylated Akt levels, but not associated with p27 and cyclin D1 expression, in primary epithelial ovarian carcinomas. AB - PTEN (MMAC1/TEP1), a tumor suppressor gene on chromosome subband 10q23.3, is variably mutated and/or deleted in a variety of human cancers. Germline mutations in PTEN, which encode a dual-specificity phosphatase, have been implicated in at least two hamartoma tumor syndromes that exhibit some clinical overlap, Cowden syndrome and Bannayan-Riley-Ruvalcaba syndrome. Among several series of ovarian cancers, the frequency of loss of heterozygosity (LOH) of markers flanking and within PTEN, is approximately 30 to 50%, and the somatic intragenic PTEN mutation frequency is <10%. In this study, we screened primary adenocarcinomas of the ovary for LOH of polymorphic markers within and flanking the PTEN gene and for intragenic mutations of the PTEN gene and compared them to PTEN expression using immunohistochemistry. Furthermore, we sought to detect the expression of the presumed downstream targets of PTEN, such as P-Akt, p27, and cyclin D1 by immunohistochemistry. LOH at 10q23 was observed in 29 of 64 (45%) cases. Of the 117 samples, 6 somatic intragenic PTEN mutations, 1 germline mutation, and 1 novel polymorphism were found in 7 (6%) patients. Immunostaining of 49 ovarian cancer samples revealed that 13 (27%) were PTEN immunostain-negative, 25 (51%) had reduced staining, and the rest (22%) were PTEN expression-positive. Among the 44 informative tumors assessed for 10q23 LOH and PTEN immunostaining, there was an association between 10q23 LOH and decreased or absent staining (P = 0.0317). Of note, there were five (11%) tumors with neither mutation nor deletion that exhibited no PTEN expression and 10 (25%) others without mutation or deletion but had decreased PTEN expression. Among the 49 tumors available for immunohistochemistry, 28 (57%) showed P-Akt-positive staining, 24 (49%) had decreased p27 staining, and cyclin D1 was overexpressed in 35 (79%) cases. In general, P-Akt expression was inversely correlated with PTEN expression (P = 0.0083). These data suggest that disruption of PTEN by several mechanisms, allelic loss, intragenic mutation, or epigenetic silencing, all contribute to epithelial ovarian carcinogenesis, and that epigenetic silencing is a significant mechanism. The Akt pathway is prominently involved, but clearly not in all cases. Surprisingly, despite in vitro demonstration that p27 and cyclin D1 lies downstream of PTEN and Akt, there was no correlation between p27 and cyclin D1 expression and PTEN or P-Akt status. Thus, in vivo, although PTEN and Akt play a prominent role in ovarian carcinogenesis, p27 and cyclin D1 might not be the primary downstream targets. Alternatively, these observations could also suggest that pathways involving other than Akt, p27 and cyclin D1 that lie downstream of PTEN play roles in ovarian carcinogenesis. PMID- 11395389 TI - Murine gammaherpesvirus-68 infection causes multi-organ fibrosis and alters leukocyte trafficking in interferon-gamma receptor knockout mice. AB - Murine gammaherpesvirus-68 (MHV-68) infection in interferon-gamma receptor knockout mice (IFN-gammaR(-)/(-)) results in splenic fibrosis and excessive loss of splenocytes. In our present study we found that MHV-68 infection in IFN gammaR(-)/(-) mice also resulted in fibrosis and atrophy of the mediastinal lymph nodes, interstitial pulmonary fibrosis and fibrotic changes in the liver. Atrophy and cellular depletion of the spleen in IFN-gammaR(-)/(-) was not the result of increased cell death. The loss of splenocytes in IFN-gammaR(-)/(-) mice, which was most evident on day 23 after infection, correlated with an increase in the number of leukocytes in peripheral blood. At the peak of leukocytosis, on day 23 after infection, peripheral blood cells from infected IFN-gammaR(-)/(-) mice were unable to traffic through the fibrosed spleens of IFN-gammaR(-)/(-) mice but were able to enter the spleens of wild-type mice. This indicates that leukocytosis was in part the result of emigration of cells from the spleen and their subsequent exclusion of re-entry at the height of fibrosis. Significant cytokine and chemokine changes were observed in spleens of IFN-gammaR(-)/(-) mice. IFN-gamma, tumor necrosis factor-alpha (TNF-alpha ), TNF-beta, interleukin-1beta (IL-1beta), transforming growth factor-beta1 (TGF-beta1), lymphotactin, and MIP-1beta were elevated on day 14 after infection whereas chemokines IP-10 and MIG were significantly reduced. These changes suggest a role for dysregulated cytokines and chemokines in severe organ-specific fibrosis with implications for immune mediated fibrotic disorders. PMID- 11395390 TI - Myelin/axonal pathology in interleukin-12 induced serial relapses of experimental allergic encephalomyelitis in the Lewis rat. AB - Lewis rats, on recovery from monophasic clinical experimental allergic encephalomyelitis (EAE), can be induced to develop repeated paralytic relapses with a graded reduction in clinical severity following intraperitoneal administration of IL-12. By the time of the third relapse, the number and size of inflammatory cuffs in the spinal cord were reduced with the makeup of the cellular infiltrate shifting to a significantly increased number of B cells. Serum levels of myelin basic protein (MBP)-specific IgG1 and IgG2b were found to rise over time while MBP and MBP peptide-positive macrophages and microglia became evident in perivascular cuffs and in spinal cord parenchyma, indicative of myelin phagocytosis. Axonal death was observed in semithin and EM sections of spinal cord in third relapse animals in association with iNOS and tPA immunostaining throughout gray and white matter. These neurotoxic or excitotoxic agents may contribute to axonal damage directly or indirectly by activated microglia and macrophages, leading to limited damage of the axonal-myelin unit. PMID- 11395391 TI - The role of metalloelastase in immune complex-induced acute lung injury. AB - Matrix metalloproteases (MMPs) are a group of zinc-dependent endopeptidases that can degrade every component of the extracellular matrix. Under normal circumstances, the levels of MMPs are tightly regulated at both transcriptional and posttranscriptional levels. However, they are up-regulated in pathological states such as inflammation. Previous investigations have suggested that MMP-12 (metalloelastase) may be an important mediator in the pathogenesis of chronic lung injury. In this study we investigated the role of metalloelastase in the pathogenesis of acute lung injury using mice containing a targeted disruption of the metalloelastase gene. Neutrophil influx into the alveolar space in metalloelastase-deficient animals was reduced to approximately 50% of that observed in parent strain mice following the induction of injury by immune complexes. In addition, lung permeability in metalloelastase-deficient mice was approximately 50% of that of injured parent strain animals with normal levels of metalloelastase and this was correlated with histological evidence of less lung injury in the metalloelastase-deficient animals. Collectively, the data suggest that metalloelastase is necessary for the full development of acute alveolitis in this model of lung injury. Further, the data suggest that reduced injury in metalloelastase-deficient mice is due in part to decreased neutrophil influx into the alveolar space. PMID- 11395392 TI - Hibernation, a model of neuroprotection. AB - Hibernation, a natural model of tolerance to cerebral ischemia, represents a state of pronounced fluctuation in cerebral blood flow where no brain damage occurs. Numerous neuroprotective aspects may contribute in concert to such tolerance. The purpose of this study was to determine whether hibernating brain tissue is tolerant to penetrating brain injury modeled by insertion of microdialysis probes. Guide cannulae were surgically implanted in striatum of Arctic ground squirrels before any of the animals began to hibernate. Microdialysis probes were then inserted in some animals after they entered hibernation and in others while they remained euthermic. The brain tissue from hibernating and euthermic animals was examined 3 days after implantation of microdialysis probes. Tissue response, indicated by examination of hematoxylin and eosin-stained tissue sections and immunocytochemical identification of activated microglia, astrocytes, and hemeoxygenase-1 immunoreactivity, was dramatically attenuated around probe tracks in hibernating animals compared to euthermic controls. No difference in tissue response around guide cannulae was observed between groups. Further study of the mechanisms underlying neuroprotective aspects of hibernation may lead to novel therapeutic strategies for stroke and traumatic brain injury. PMID- 11395393 TI - Energetic determinants of tyrosine phosphorylation of focal adhesion proteins during hypoxia/reoxygenation of kidney proximal tubules. AB - Anaerobic mitochondrial metabolism of alpha-ketoglutarate and aspartate or alpha ketoglutarate and malate can prevent and reverse severe mitochondrial dysfunction during reoxygenation after 60 minutes of hypoxia in kidney proximal tubules.(34) The present studies demonstrate that, during hypoxia, paxillin, focal adhesion kinase, and p130(cas) migrated faster by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, their phosphotyrosine (pY) content decreased to approximately 5% of that in oxygenated tubules without changes in total protein, and the normally basal immunostaining of beta1 and alpha6 integrin subunits, pY, and paxillin was lost or markedly decreased. During reoxygenation without supplemental substrates, recovery of pY and basal localization of the focal adhesion proteins was poor. alpha-Ketoglutarate and aspartate, which maintained slightly higher levels of ATP during hypoxia, also maintained 2.5-fold higher levels of pY during this period, and promoted full recovery of pY content and basal localization of focal adhesion proteins during subsequent reoxygenation. Similarly complete recovery was made possible by provision of alpha-ketoglutarate and aspartate or alpha-ketoglutarate and malate only during reoxygenation. These data emphasize the importance of very low energy thresholds for maintaining the integrity of key structural and biochemical components required for cellular survival and reaffirm the value of approaches aimed at conserving or generating energy in cells injured by hypoxia or ischemia. PMID- 11395394 TI - Amyloid angiopathy and variability in amyloid beta deposition is determined by mutation position in presenilin-1-linked Alzheimer's disease. AB - The presenilins (PSs) are components of large molecular complexes that contain beta-catenin and function as gamma-secretase. We report here a striking correlation between amyloid angiopathy and the location of mutation in PS-1 linked Alzheimer's disease. The amount of amyloid beta protein, Abeta(42(43)), but not Abeta(40,) deposited in the frontal cortex of the brain is increased in 54 cases of early-onset familial Alzheimer's disease, encompassing 25 mutations in the presenilin-1 (PS-1) gene, compared to sporadic Alzheimer's disease. The amount of Abeta(40) in PS-1 Alzheimer's disease varied according to the copy number of epsilon4 alleles of the Apolipoprotein E gene. Although the amounts of Abeta(40) and Abeta(42(43)) deposited did not correlate with the genetic location of the mutation in a strict linear sense, the histological profile did so vary. Cases with mutations between codon 1 and 200 showed, in frontal cortex, many diffuse plaques, few cored plaques, and mild or moderate amyloid angiopathy. Cases with mutations occurring after codon 200 also showed many diffuse plaques, but the number and size of cored plaques were increased (even when epsilon4 allele was not present) and these were often clustered around blood vessels severely affected by amyloid angiopathy. Similarly, diverging histological profiles, mainly according to the degree of amyloid angiopathy, were seen in the cerebellum. Mutations in the PS-1 gene may therefore alter the topology of the PS 1 protein so as to favor Abeta formation and deposition, generally, but also to facilitate amyloid angiopathy particularly in cases in which the mutation lies beyond codon 200. Finally we report that the amount of Abeta(42(43)) deposited in the brain correlated with the amount of this produced in culture by cells bearing the equivalent mutations. PMID- 11395396 TI - The NPM-ALK and the ATIC-ALK fusion genes can be detected in non-neoplastic cells. AB - Anaplastic large cell lymphoma (ALCL) is frequently associated with the t(2;5)(p23;q35) translocation. It creates a NPM-ALK fusion gene, fusing the anaplastic lymphoma kinase (ALK) gene (2p23) and the nucleophosmin (NPM) gene (5q35). Other rearrangements involving the ALK gene have recently been shown to be associated with ALCL, among which the ATIC-ALK rearrangement resulting from the inv(2)(p23q35) translocation is probably the most recurrent. The aims of the present study were to investigate the presence of NPM-ALK and ATIC-ALK fusion genes in ALCL, using a real-time 5' exonuclease-based reverse-transcription polymerase chain reaction (RT-PCR). This sensitive technique was also applied to investigate whether both fusion genes might be detected in Hodgkin's disease cases and in reactive lymphoid tissue. Results of the RT-PCR were compared to ALK immunostaining, cytogenetics, and fluorescence in situ hybridization (FISH) results. RT-PCR detected the NPM-ALK and ATIC-ALK fusions at high levels in 8 and 3 of a total of 13 ALK-positive ALCL cases. One ALK-positive ALCL case was negative for both fusion genes analyzed but revealed a new ALK-related translocation t(2;17)(p23;q25) by cytogenetic and FISH analysis. In addition, of the eight ALK-positive ALCL cases that were strongly positive for the NPM-ALK fusion, three cases also showed the presence of the ATIC-ALK fusion, although at much lower levels. Similarly, out of the three strongly positive ATIC-ALK cases, one case was positive for the NPM-ALK fusion, at low levels. Finally, the NPM-ALK and the ATIC-ALK fusions were detected, at equally low levels, respectively in 13 and 5 ALK-negative ALCL cases, in 11 and 5 Hodgkin's disease cases and in 20 and 1 non-neoplastic lymphoid tissues. The distinction between the high- and low level detection was confirmed by relative quantitative RT-PCR for a representative number of cases. Of interest is the fact that the high-level detection coincided with the presence of ALK gene rearrangement detected by cytogenetics and FISH and may reflect a central role of the transcript in the oncogenic mechanism of ALK-positive ALCL. Low-level detection is not supported by cytogenetics and FISH, presumably due to the presence of the transcripts in only a small minority of normal cells not detectable by these techniques. Our findings demonstrate that NPM-ALK and ATIC-ALK fusion transcripts may be detected in conditions other than ALK-positive ALCL including reactive lymphoid tissues, although at low levels, suggesting the presence of the transcripts in normal (bystander) cells. Moreover, they suggest that the ALK gene rearrangement by itself might be insufficient to induce tumor formation. They further question the validity of quantitative real-time RT-PCR for monitoring minimal residual disease in ALCL. Finally, the newly identified translocation t(2;17)(p23;q25) can be added to the list of ALK gene rearrangements occurring in ALK-positive ALCL. PMID- 11395395 TI - Unbalanced expression of 11p15 imprinted genes in focal forms of congenital hyperinsulinism: association with a reduction to homozygosity of a mutation in ABCC8 or KCNJ11. AB - Congenital hyperinsulinism (CHI), previously named persistent hyperinsulinemic hypoglycemia of infancy, is characterized by profound hypoglycemia because of excessive insulin secretion. CHI presents as two different morphological forms: a diffuse form with functional abnormality of islets throughout the pancreas and a focal form with focal islet cell adenomatous hyperplasia, which can be cured by partial pancreatectomy. Recently, we have shown that focal adenomatous hyperplasia involves the specific loss of the maternal 11p15 region and a constitutional mutation of a paternally inherited allele of the gene encoding the regulating subunit of the K(+)(ATP) channel, the sulfonylurea receptor (ABCC8 or SUR1). In the present study on a large series of 31 patients, describing both morphological features and molecular data, we report that 61% of cases (19 out of 31) carried a paternally inherited mutation not only in the ABCC8 gene as previously described but also in the second gene encoding the K(+)(ATP) channel, the inward rectifying potassium channel (KCNJ11 or KIR6.2), in 15 cases and 4 cases, respectively. Moreover our results are consistent with the presence of a duplicated paternal 11p15 allele probably because of mitotic recombination or reduplication of the paternal chromosome after somatic loss of the maternal chromosome. In agreement with the loss of the maternal chromosome, the level of expression of a maternally expressed tumor suppressor gene, H19, was greatly reduced compared to the level of expression of the paternally expressed growth promoter gene, IGF2. The expression of IGF2 was on average only moderately increased. Thus, focal forms of CHI can be considered to be a recessive somatic disease, associating an imbalance in the expression of imprinted genes in the 11p15.5 region to a somatic reduction to homozygosity of an ABCC8- or KCNJ11 recessive mutation. The former is responsible for the abnormal growth rate, as in embryonic tumors, whereas the latter leads to unregulated secretion of insulin. PMID- 11395397 TI - A quantitative model for the dynamics of serum prostate-specific antigen as a marker for cancerous growth: an explanation for a medical anomaly. AB - Prostate-specific antigen (PSA) is an enzyme produced by both normal and cancerous prostate epithelial cells. Although PSA is the most widely used serum marker to detect and follow patients with prostatic adenocarcinoma, there are certain anomalies in the values of serum levels of PSA that are not understood. We developed a mathematical model for the dynamics of serum levels of PSA as a function of the tumor volume. Our model results show good agreement with experimental observations and provide an explanation for the existence of significant prostatic tumor mass despite a low-serum PSA. This result can be very useful in enhancing the use of serum PSA levels as a marker for cancer growth. PMID- 11395398 TI - Prion proteins with different conformations accumulate in Gerstmann-Straussler Scheinker disease caused by A117V and F198S mutations. AB - Gerstmann-Straussler-Scheinker disease (GSS) is characterized by the accumulation of proteinase K (PK)-resistant prion protein fragments (PrP(sc)) of approximately 7 to 15 kd in the brain. Purified GSS amyloid is composed primarily of approximately 7-kd PrP peptides, whose N terminus corresponds to residues W(81) and G(88) to G(90) in patients with the A117V mutation and to residue W(81) in patients with the F198S mutation. The aim of this study was to characterize PrP in brain extracts, microsomal preparations, and purified fractions from A117V patients and to determine the N terminus of PrP(sc) species in both GSS A117V and F198S. In all GSS A117V patients, the approximately 7-kd PrP(sc) fragment isolated from nondigested and PK-digested samples had the major N terminus at residue G(88) and G(90), respectively. Conversely, in all patients with GSS F198S, an approximately 8-kd PrP(sc) fragment was isolated having the major N terminus start at residue G(74). It is possible that a further degradation of this fragment generates the amyloid subunit starting at W(81). The finding that patients with GSS A117V and F198S accumulate PrP(sc) fragments of different size and N-terminal sequence, suggests that these mutations generate two distinct PrP conformers. PMID- 11395399 TI - Accumulation of amyloid beta-protein in the low-density membrane domain accurately reflects the extent of beta-amyloid deposition in the brain. AB - To learn more about the process of amyloid beta-protein (Abeta) deposition in the brain, human prefrontal cortices were fractionated by sucrose density gradient centrifugation, and the Abeta content in each fraction was quantified by a two site enzyme-linked immunosorbent assay. The fractionation protocol revealed two pools of insoluble Abeta. One corresponded to a low-density membrane domain; the other was primarily composed of extracellular Abeta deposits in those cases in which Abeta accumulated to significant levels. Abeta42 levels in the low-density membrane domain were proportional to the extent of total Abeta42 accumulation, which is known to correlate well with overall amyloid burden. In PDAPP mice that form senile plaques and accumulate Abeta in a similar manner to aging humans, Abeta42 accumulation in the low-density membrane domain also increased as Abeta deposition progressed with aging. These observations indicate that the Abeta42 associated with low-density membrane domains is tightly coupled with the process of extracellular Abeta deposition. PMID- 11395400 TI - Humoral hypercalcemia of malignancy: severe combined immunodeficient/beige mouse model of adult T-cell lymphoma independent of human T-cell lymphotropic virus type-1 tax expression. AB - The majority of patients with adult T-cell leukemia/lymphoma (ATL) resulting from human T-cell lymphotropic virus type-1 (HTLV-1) infection develop humoral hypercalcemia of malignancy (HHM). We used an animal model using severe combined immunodeficient (SCID)/beige mice to study the pathogenesis of HHM. SCID/beige mice were inoculated intraperitoneally with a human ATL line (RV-ATL) and were euthanized 20 to 32 days after inoculation. SCID/beige mice with engrafted RV-ATL cells developed lymphoma in the mesentery, liver, thymus, lungs, and spleen. The lymphomas stained positively for human CD45RO surface receptor and normal mouse lymphocytes stained negatively confirming the human origin of the tumors. The ATL cells were immunohistochemically positive for parathyroid hormone-related protein (PTHrP). In addition, PTHrP mRNA was highly expressed in lymphomas when compared to MT-2 cells (HTLV-1-positive cell line). Mice with lymphoma developed severe hypercalcemia. Plasma PTHrP concentrations were markedly increased in mice with hypercalcemia, and correlated with the increase in plasma calcium concentrations. Bone densitometry and histomorphometry in lymphoma-bearing mice revealed significant bone loss because of a marked increase in osteoclastic bone resorption. RV-ATL cells contained 1.5 HTLV-1 proviral copies of the tax gene as determined by quantitative real-time polymerase chain reaction (PCR). However, tax expression was not detected by Western blot or reverse transcriptase (RT)-PCR in RV-ATL cells, which suggests that factors other than Tax are modulators of PTHrP gene expression. The SCID/beige mouse model mimics HHM as it occurs in ATL patients, and will be useful to investigate the regulation of PTHrP expression by ATL cells in vivo. PMID- 11395401 TI - Advancing our knowledge in biochemistry, genetics, and microbiology through studies on tryptophan metabolism. AB - I was fortunate to practice science during the last half of the previous century, when many basic biological and biochemical concepts could be experimentally addressed for the first time. My introduction to research involved isolating and identifying intermediates in the niacin biosynthetic pathway. These studies were followed by investigations focused on determining the properties of genes and enzymes essential to metabolism and examining how they were alterable by mutation. The most challenging problem I initially attacked was establishing the colinear relationship between gene and protein. Subsequent research emphasized identification and characterization of regulatory mechanisms that microorganisms use to control gene expression. An elaborate regulatory strategy, transcription attenuation, was discovered that is often based on selection between alternative RNA structures. Throughout my career I enjoyed the excitement of solving basic scientific problems. Most rewarding, however, was the feeling that I was helping young scientists experience the pleasure of performing creative research. PMID- 11395402 TI - DNA primases. AB - DNA primases are enzymes whose continual activity is required at the DNA replication fork. They catalyze the synthesis of short RNA molecules used as primers for DNA polymerases. Primers are synthesized from ribonucleoside triphosphates and are four to fifteen nucleotides long. Most DNA primases can be divided into two classes. The first class contains bacterial and bacteriophage enzymes found associated with replicative DNA helicases. These prokaryotic primases contain three distinct domains: an amino terminal domain with a zinc ribbon motif involved in binding template DNA, a middle RNA polymerase domain, and a carboxyl-terminal region that either is itself a DNA helicase or interacts with a DNA helicase. The second major primase class comprises heterodimeric eukaryotic primases that form a complex with DNA polymerase alpha and its accessory B subunit. The small eukaryotic primase subunit contains the active site for RNA synthesis, and its activity correlates with DNA replication during the cell cycle. PMID- 11395403 TI - Histone acetyltransferases. AB - Transcriptional regulation in eukaryotes occurs within a chromatin setting and is strongly influenced by nucleosomal barriers imposed by histone proteins. Among the well-known covalent modifications of histones, the reversible acetylation of internal lysine residues in histone amino-terminal domains has long been positively linked to transcriptional activation. Recent biochemical and genetic studies have identified several large, multisubunit enzyme complexes responsible for bringing about the targeted acetylation of histones and other factors. This review discusses our current understanding of histone acetyltransferases (HATs) or acetyltransferases (ATs): their discovery, substrate specificity, catalytic mechanism, regulation, and functional links to transcription, as well as to other chromatin-modifying activities. Recent studies underscore unexpected connections to both cellular regulatory processes underlying normal development and differentiation, as well as abnormal processes that lead to oncogenesis. Although the functions of HATs and the mechanisms by which they are regulated are only beginning to be understood, these fundamental processes are likely to have far reaching implications for human biology and disease. PMID- 11395404 TI - Radical mechanisms of enzymatic catalysis. AB - Two classes of enzymatic mechanisms that proceed by free radical chemistry initiated by the 5'-deoxyadenosyl radical are discussed. In the first class, the mechanism of the interconversion of L-lysine and L-beta-lysine catalyzed by lysine 2,3-aminomutase (LAM) involves four radicals, three of which have been spectroscopically characterized. The reversible formation of the 5'-deoxyadenosyl radical takes place by the chemical cleavage of S-adenosylmethionine (SAM) reacting with the [4Fe-4S]+ center in LAM. In other reactions of SAM with iron sulfur proteins, SAM is irreversibly consumed to generate the 5'-deoxyadenosyl radical, which activates an enzyme by abstracting a hydrogen atom from an enzymatic glycyl residue to form a glycyl radical. The glycyl radical enzymes include pyruvate formate-lyase, anaerobic ribonucleotide reductase from Escherichia coli, and benzylsuccinate synthase. Biotin synthase and lipoate synthase are SAM-dependent [4Fe-4S] proteins that catalyze the insertion of sulfur into unactivated C-H bonds, which are cleaved by the 5'-deoxyadenosyl radical from SAM. In the second class of enzymatic mechanisms using free radicals, adenosylcobalamin-dependent reactions, the 5'-deoxyadenosyl radical arises from homolytic cleavage of the cobalt-carbon bond, and it initiates radical reactions by abstracting hydrogen atoms from substrates. Three examples are described of suicide inactivation through the formation of exceptionally stable free radicals at enzymatic active sites. PMID- 11395405 TI - Channeling of substrates and intermediates in enzyme-catalyzed reactions. AB - The three-dimensional structures of tryptophan synthase, carbamoyl phosphate synthetase, glutamine phosphoribosylpyrophosphate amidotransferase, and asparagine synthetase have revealed the relative locations of multiple active sites within these proteins. In all of these polyfunctional enzymes, a product formed from the catalytic reaction at one active site is a substrate for an enzymatic reaction at a distal active site. Reaction intermediates are translocated from one active site to the next through the participation of an intermolecular tunnel. The tunnel in tryptophan synthase is approximately 25 A in length, whereas the tunnel in carbamoyl phosphate synthetase is nearly 100 A long. Kinetic studies have demonstrated that the individual reactions are coordinated through allosteric coupling of one active site with another. The participation of these molecular tunnels is thought to protect reactive intermediates from coming in contact with the external medium. PMID- 11395406 TI - Replisome-mediated DNA replication. AB - The elaborate process of genomic replication requires a large collection of proteins properly assembled at a DNA replication fork. Several decades of research on the bacterium Escherichia coli and its bacteriophages T4 and T7 have defined the roles of many proteins central to DNA replication. These three different prokaryotic replication systems use the same fundamental components for synthesis at a moving DNA replication fork even though the number and nature of some individual proteins are different and many lack extensive sequence homology. The components of the replication complex can be grouped into functional categories as follows: DNA polymerase, helix destabilizing protein, polymerase accessory factors, and primosome (DNA helicase and DNA primase activities). The replication of DNA derives from a multistep enzymatic pathway that features the assembly of accessory factors and polymerases into a functional holoenzyme; the separation of the double-stranded template DNA by helicase activity and its coupling to the primase synthesis of RNA primers to initiate Okazaki fragment synthesis; and the continuous and discontinuous synthesis of the leading and lagging daughter strands by the polymerases. This review summarizes and compares and contrasts for these three systems the types, timing, and mechanism of reactions and of protein-protein interactions required to initiate, control, and coordinate the synthesis of the leading and lagging strands at a DNA replication fork and comments on their generality. PMID- 11395407 TI - Divergent evolution of enzymatic function: mechanistically diverse superfamilies and functionally distinct suprafamilies. AB - The protein sequence and structure databases are now sufficiently representative that strategies nature uses to evolve new catalytic functions can be identified. Groups of divergently related enzymes whose members catalyze different reactions but share a common partial reaction, intermediate, or transition state (mechanistically diverse superfamilies) have been discovered, including the enolase, amidohydrolase, thiyl radical, crotonase, vicinal-oxygen-chelate, and Fe dependent oxidase superfamilies. Other groups of divergently related enzymes whose members catalyze different overall reactions that do not share a common mechanistic strategy (functionally distinct suprafamilies) have also been identified: (a) functionally distinct suprafamilies whose members catalyze successive transformations in the tryptophan and histidine biosynthetic pathways and (b) functionally distinct suprafamilies whose members catalyze different reactions in different metabolic pathways. An understanding of the structural bases for the catalytic diversity observed in super- and suprafamilies may provide the basis for discovering the functions of proteins and enzymes in new genomes as well as provide guidance for in vitro evolution/engineering of new enzymes. PMID- 11395408 TI - PTEN and myotubularin: novel phosphoinositide phosphatases. AB - Protein tyrosine phosphatases (PTPs) are a diverse group of enzymes that contain a highly conserved active site motif, Cys-x5-Arg (Cx5R). The PTP superfamily enzymes, which include tyrosine-specific, dual specificity, low-molecular-weight, and Cdc25 phosphatases, are key mediators of a wide variety of cellular processes, including growth, metabolism, differentiation, motility, and programmed cell death. The PTEN/MMAC1/TEP1 gene was originally identified as a candidate tumor suppressor gene located on human chromosome 10q23; it encodes a protein with sequence similarity to PTPs and tensin. Recent studies have demonstrated that PTEN plays an essential role in regulating signaling pathways involved in cell growth and apoptosis, and mutations in the PTEN gene are now known to cause tumorigenesis in a number of human tissues. In addition, germ line mutations in the PTEN gene also play a major role in the development of Cowden and Bannayan-Zonana syndromes, in which patients often suffer from increased risk of breast and thyroid cancers. Biochemical studies of the PTEN phosphatase have revealed a molecular mechanism by which tumorigenesis may be caused in individuals with PTEN mutations. Unlike most members of the PTP superfamily, PTEN utilizes the phosphoinositide second messenger, phosphatidylinositol 3,4,5 trisphosphate (PIP3), as its physiologic substrate. This inositol lipid is an important regulator of cell growth and survival signaling through the Ser/Thr protein kinases PDK1 and Akt. By specifically dephosphorylating the D3 position of PIP3, the PTEN tumor suppressor functions as a negative regulator of signaling processes downstream of this lipid second messenger. Mutations that impair PTEN function result in a marked increase in cellular levels of PIP3 and constitutive activation of Akt survival signaling pathways, leading to inhibition of apoptosis, hyperplasia, and tumor formation. Certain structural features of PTEN contribute to its specificity for PIP3, as well as its role(s) in regulating cellular proliferation and apoptosis. Recently, myotubularin, a second PTP superfamily enzyme associated with human disease, has also been shown to utilize a phosphoinositide as its physiologic substrate. PMID- 11395409 TI - Regulation of phosphoinositide-specific phospholipase C. AB - Eleven distinct isoforms of phosphoinositide-specific phospholipase C (PLC), which are grouped into four subfamilies (beta, gamma, delta, and epsilon), have been identified in mammals. These isozymes catalyze the hydrolysis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] to inositol 1,4,5 trisphosphate and diacylglycerol in response to the activation of more than 100 different cell surface receptors. All PLC isoforms contain X and Y domains, which form the catalytic core, as well as various combinations of regulatory domains that are common to many other signaling proteins. These regulatory domains serve to target PLC isozymes to the vicinity of their substrate or activators through protein-protein or protein-lipid interactions. These domains (with their binding partners in parentheses or brackets) include the pleckstrin homology (PH) domain [PtdIns(3)P, beta gamma subunits of G proteins] and the COOH-terminal region including the C2 domain (GTP-bound alpha subunit of Gq) of PLC-beta; the PH domain [PtdIns(3,4,5)P3] and Src homology 2 domain [tyrosine-phosphorylated proteins, PtdIns(3,4,5)P3] of PLC-gamma; the PH domain [PtdIns(4,5)P2] and C2 domain (Ca2+) of PLC-delta; and the Ras binding domain (GTP-bound Ras) of PLC epsilon. The presence of distinct regulatory domains in PLC isoforms renders them susceptible to different modes of activation. Given that the partners that interact with these regulatory domains of PLC isozymes are generated or eliminated in specific regions of the cell in response to changes in receptor status, the activation and deactivation of each PLC isoform are likely highly regulated processes. PMID- 11395410 TI - Design and selection of novel Cys2His2 zinc finger proteins. AB - Cys2His2 zinc finger proteins offer a stable and versatile framework for the design of proteins that recognize desired target sites on double-stranded DNA. Individual fingers from these proteins have a simple beta beta alpha structure that folds around a central zinc ion, and tandem sets of fingers can contact neighboring subsites of 3-4 base pairs along the major groove of the DNA. Although there is no simple, general code for zinc finger-DNA recognition, selection strategies have been developed that allow these proteins to be targeted to almost any desired site on double-stranded DNA. The affinity and specificity of these new proteins can also be improved by linking more fingers together or by designing proteins that bind as dimers and thus recognize an extended site. These new proteins can then be modified by adding other domains--for activation or repression of transcription, for DNA cleavage, or for other activities. Such designer transcription factors and other new proteins will have important applications in biomedical research and in gene therapy. PMID- 11395411 TI - Peroxisome proliferator-activated receptor gamma and metabolic disease. AB - The nuclear peroxisome proliferator-activated receptor gamma (PPAR gamma) is a transcription factor that is activated by polyunsaturated fatty acids and their metabolites and is essential for fat cell formation. Although obesity is a strong risk factor for type 2 diabetes mellitus and other metabolic diseases, potent PPAR gamma activators such as the glitazone drugs lower glucose and lipid levels in patients with type 2 diabetes and also have antiatherosclerotic and antihypertensive effects. We review recent studies providing insight into the paradoxical relationship between PPAR gamma and metabolic disease. We also review recent advances in understanding the structural basis for PPAR gamma activation by ligands. The unusual ligand-binding properties of PPAR gamma suggest that it will be possible to discover new chemical classes of receptor "modulators" with distinct pharmacological activities for the treatment of type 2 diabetes and other metabolic diseases. PMID- 11395412 TI - DNA topoisomerases: structure, function, and mechanism. AB - DNA topoisomerases solve the topological problems associated with DNA replication, transcription, recombination, and chromatin remodeling by introducing temporary single- or double-strand breaks in the DNA. In addition, these enzymes fine-tune the steady-state level of DNA supercoiling both to facilitate protein interactions with the DNA and to prevent excessive supercoiling that is deleterious. In recent years, the crystal structures of a number of topoisomerase fragments, representing nearly all the known classes of enzymes, have been solved. These structures provide remarkable insights into the mechanisms of these enzymes and complement previous conclusions based on biochemical analyses. Surprisingly, despite little or no sequence homology, both type IA and type IIA topoisomerases from prokaryotes and the type IIA enzymes from eukaryotes share structural folds that appear to reflect functional motifs within critical regions of the enzymes. The type IB enzymes are structurally distinct from all other known topoisomerases but are similar to a class of enzymes referred to as tyrosine recombinases. The structural themes common to all topoisomerases include hinged clamps that open and close to bind DNA, the presence of DNA binding cavities for temporary storage of DNA segments, and the coupling of protein conformational changes to DNA rotation or DNA movement. For the type II topoisomerases, the binding and hydrolysis of ATP further modulate conformational changes in the enzymes to effect changes in DNA topology. PMID- 11395413 TI - Fidelity of aminoacyl-tRNA selection on the ribosome: kinetic and structural mechanisms. AB - The ribosome discriminates between correct and incorrect aminoacyl-tRNAs (aa tRNAs), or their complexes with elongation factor Tu (EF-Tu) and GTP, according to the match between anticodon and mRNA codon in the A site. Selection takes place at two stages, prior to GTP hydrolysis (initial selection) and after GTP hydrolysis but before peptide bond formation (proofreading). In part, discrimination results from different rejection rates that are due to different stabilities of the respective codon-anticodon complexes. An important additional contribution is provided by induced fit, in that only correct codon recognition leads to acceleration of rate-limiting rearrangements that precede chemical steps. Recent elucidation of ribosome structures and mutational analyses suggest which residues of the decoding center may be involved in signaling formation of the correct codon-anticodon duplex to the functional centers of the ribosome. In utilizing induced fit for substrate discrimination, the ribosome resembles other nucleic acid-programmed polymerases. PMID- 11395414 TI - Analysis of proteins and proteomes by mass spectrometry. AB - A decade after the discovery of electrospray and matrix-assisted laser desorption ionization (MALDI), methods that finally allowed gentle ionization of large biomolecules, mass spectrometry has become a powerful tool in protein analysis and the key technology in the emerging field of proteomics. The success of mass spectrometry is driven both by innovative instrumentation designs, especially those operating on the time-of-flight or ion-trapping principles, and by large scale biochemical strategies, which use mass spectrometry to detect the isolated proteins. Any human protein can now be identified directly from genome databases on the basis of minimal data derived by mass spectrometry. As has already happened in genomics, increased automation of sample handling, analysis, and the interpretation of results will generate an avalanche of qualitative and quantitative proteomic data. Protein-protein interactions can be analyzed directly by precipitation of a tagged bait followed by mass spectrometric identification of its binding partners. By these and similar strategies, entire protein complexes, signaling pathways, and whole organelles are being characterized. Posttranslational modifications remain difficult to analyze but are starting to yield to generic strategies. PMID- 11395415 TI - Transcriptional coactivator complexes. AB - The last two decades have witnessed a tremendous expansion in our knowledge of the mechanisms employed by eukaryotic cells to control gene activity. A critical insight to transcriptional control mechanisms was provided by the discovery of coactivators, a diverse array of cellular factors that connect sequence-specific DNA binding activators to the general transcriptional machinery, or that help activators and the transcriptional apparatus to navigate through the constraints of chromatin. A number of coactivators have been isolated as large multifunctional complexes, and biochemical, genetic, molecular, and cellular strategies have all contributed to uncovering many of their components, activities, and modes of action. Coactivator functions can be broadly divide into two classes: (a) adaptors that direct activator recruitment of the transcriptional apparatus, (b) chromatin-remodeling or -modifying enzymes. Strikingly, several distinct coactivator complexes nonetheless share many subunits and appear to be assembled in a modular fashion. Such structural and functional modularity could provide the cell with building blocks from which to construct a versatile array of coactivator complexes according to its needs. The extent of functional interplay between these different activities in gene specific transcriptional regulation is only now becoming apparent, and will remain an active area of research for years to come. PMID- 11395416 TI - Mechanisms underlying ubiquitination. AB - The conjugation of ubiquitin to other cellular proteins regulates a broad range of eukaryotic cell functions. The high efficiency and exquisite selectivity of ubiquitination reactions reflect the properties of enzymes known as ubiquitin protein ligases or E3s. An E3 recognizes its substrates based on the presence of a specific ubiquitination signal, and catalyzes the formation of an isopeptide bond between a substrate (or ubiquitin) lysine residue and the C terminus of ubiquitin. Although a great deal is known about the molecular basis of E3 specificity, much less is known about molecular mechanisms of catalysis by E3s. Recent findings reveal that all known E3s utilize one of just two catalytic domains--a HECT domain or a RING finger--and crystal structures have provided the first detailed views of an active site of each type. The new findings shed light on many aspects of E3 structure, function, and mechanism, but also emphasize that key features of E3 catalysis remain to be elucidated. PMID- 11395417 TI - Synthesis and function of 3-phosphorylated inositol lipids. AB - The 3-phosphorylated inositol lipids fulfill roles as second messengers by interacting with the lipid binding domains of a variety of cellular proteins. Such interactions can affect the subcellular localization and aggregation of target proteins, and through allosteric effects, their activity. Generation of 3 phosphoinositides has been documented to influence diverse cellular pathways and hence alter a spectrum of fundamental cellular activities. This review is focused on the 3-phosphoinositide lipids, the synthesis of which is acutely triggered by extracellular stimuli, the enzymes responsible for their synthesis and metabolism, and their cell biological roles. Much knowledge has recently been gained through structural insights into the lipid kinases, their interaction with inhibitors, and the way their 3-phosphoinositide products interact with protein targets. This field is now moving toward a genetic dissection of 3 phosphoinositide action in a variety of model organisms. Such approaches will reveal the true role of the 3-phosphoinositides at the organismal level in health and disease. PMID- 11395418 TI - Folding of newly translated proteins in vivo: the role of molecular chaperones. AB - Recent years have witnessed dramatic advances in our understanding of how newly translated proteins fold in the cell and the contribution of molecular chaperones to this process. Folding in the cell must be achieved in a highly crowded macromolecular environment, in which release of nonnative polypeptides into the cytosolic solution might lead to formation of potentially toxic aggregates. Here I review the cellular mechanisms that ensure efficient folding of newly translated proteins in vivo. De novo protein folding appears to occur in a protected environment created by a highly processive chaperone machinery that is directly coupled to translation. Genetic and biochemical analysis shows that several distinct chaperone systems, including Hsp70 and the cylindrical chaperonins, assist the folding of proteins upon translation in the cytosol of both prokaryotic and eukaryotic cells. The cellular chaperone machinery is specifically recruited to bind to ribosomes and protects nascent chains and folding intermediates from nonproductive interactions. In addition, initiation of folding during translation appears to be important for efficient folding of multidomain proteins. PMID- 11395419 TI - Regulation of actin filament network formation through ARP2/3 complex: activation by a diverse array of proteins. AB - Actin filament assembly and turnover drive many forms of cellular motility, particularly extension of the leading edge of locomoting cells and rocketing of pathogenic microorganisms through host cell cytoplasm. De novo nucleation of actin filaments appears to be required for these movements. A complex of seven proteins called Arp2/3 complex is the best characterized cellular initiator of actin filament nucleation. Arp2/3 complex is intrinsically inactive, relying on nucleation promoting factors for activation. WASp/Scar family proteins are prominent cellular nucleation promoting factors. They bring together an actin monomer and Arp2/3 complex in solution or on the side of an existing actin filament to initiate a new filament that grows in the barbed end direction. WASp and N-WASP are intrinsically autoinhibited, and their activity is regulated by Rho-family GTPases such as Cdc42, membrane polyphosphoinositides, WIP/verprolin, and SH3 domain proteins. These interactions provide a final common pathway for many signaling inputs to regulate actin polymerization. Microorganisms either activate Arp2/3 complex directly or usurp N-WASP to initiate actin polymerization. PMID- 11395420 TI - Function, structure, and mechanism of intracellular copper trafficking proteins. AB - Genetic, biochemical, and spectroscopic studies have established a new function for an intracellular protein, i.e., guiding and inserting a copper cofactor into the active site of a target enzyme. Studies of these new proteins have revealed a fundamental aspect of copper physiology, namely the vast overcapacity of the cytoplasm for copper sequestration. This finding framed the mechanistic, energetic, and structural aspects of intracellular copper trafficking proteins. One hallmark of the copper chaperones is the similarity of the protein fold between the chaperone and its target enzyme. The surface residues presented by each partner, however, are quite different, and some initial findings concerning the complementarity of these interfaces have led to mechanistic insights. The copper chaperones appear to lower the activation barrier for metal transfer into specific protein-binding sites. The manner in which they facilitate metal insertion appears to involve a docking of the metal donor and acceptor sites in close proximity to one another. Although the intimate mechanism is still open, it appears that a low activation barrier for metal transfer is achieved by a network of coordinate-covalent, electrostatic, and hydrogen bonding interactions in the vicinity of the metal-binding site itself. PMID- 11395421 TI - Regulation of G protein-initiated signal transduction in yeast: paradigms and principles. AB - All cells have the capacity to evoke appropriate and measured responses to signal molecules (such as peptide hormones), environmental changes, and other external stimuli. Tremendous progress has been made in identifying the proteins that mediate cellular response to such signals and in elucidating how events at the cell surface are linked to subsequent biochemical changes in the cytoplasm and nucleus. An emerging area of investigation concerns how signaling components are assembled and regulated (both spatially and temporally), so as to control properly the specificity and intensity of a given signaling pathway. A related question under intensive study is how the action of an individual signaling pathway is integrated with (or insulated from) other pathways to constitute larger networks that control overall cell behavior appropriately. This review describes the signal transduction pathway used by budding yeast (Saccharomyces cerevisiae) to respond to its peptide mating pheromones. This pathway is comprised by receptors, a heterotrimeric G protein, and a protein kinase cascade all remarkably similar to counterparts in multicellular organisms. The primary focus of this review, however, is recent advances that have been made, using primarily genetic methods, in identifying molecules responsible for regulation of the action of the components of this signaling pathway. Just as many of the constituent proteins of this pathway and their interrelationships were first identified in yeast, the functions of some of these regulators have clearly been conserved in metazoans, and others will likely serve as additional models for molecules that carry out analogous roles in higher organisms. PMID- 11395422 TI - The signal recognition particle. AB - The signal recognition particle (SRP) and its membrane-associated receptor (SR) catalyze targeting of nascent secretory and membrane proteins to the protein translocation apparatus of the cell. Components of the SRP pathway and salient features of the molecular mechanism of SRP-dependent protein targeting are conserved in all three kingdoms of life. Recent advances in the structure determination of a number of key components in the eukaryotic and prokaryotic SRP pathway provide new insight into the molecular basis of SRP function, and they set the stage for future work toward an integrated picture that takes into account the dynamic and contextual properties of this remarkable cellular machine. PMID- 11395424 TI - ExScript: an 'ex'-centric approach to the description of transcript diversity. PMID- 11395423 TI - Mechanisms of viral membrane fusion and its inhibition. AB - Viral envelope glycoproteins promote viral infection by mediating the fusion of the viral membrane with the host-cell membrane. Structural and biochemical studies of two viral glycoproteins, influenza hemagglutinin and HIV-1 envelope protein, have led to a common model for viral entry. The fusion mechanism involves a transient conformational species that can be targeted by therapeutic strategies. This mechanism of infectivity is likely utilized by a wide variety of enveloped viruses for which similar therapeutic interventions should be possible. PMID- 11395425 TI - Uniform integration of genome mapping data using intersection graphs. AB - MOTIVATION: The methods for analyzing overlap data are distinct from those for analyzing probe data, making integration of the two forms awkward. Conversion of overlap data to probe-like data elements would facilitate comparison and uniform integration of overlap data and probe data using software developed for analysis of STS data. RESULTS: We show that overlap data can be effectively converted to probe-like data elements by extracting maximal sets of mutually overlapping clones. We call these sets virtual probes, since each set determines a site in the genome corresponding to the region which is common among the clones of the set. Finding the virtual probes is equivalent to finding the maximal cliques of a graph. We modify a known maximal-clique algorithm such that it finds all virtual probes in a large dataset within minutes. We illustrate the algorithm by converting fingerprint and Alu-PCR overlap data to virtual probes. The virtual probes are then analyzed using double-linkage intersection graphs and structure graphs to show that methods designed for STS data are also applicable to overlap data represented as virtual probes. Next we show that virtual probes can produce a uniform integration of different kinds of mapping data, in particular STS probe data and fingerprint and Alu-PCR overlap data. The integrated virtual probes produce longer double-linkage contigs than STS probes alone, and in conjunction with structure graphs they facilitate the identification and elimination of anomalies. Thus, the virtual-probe technique provides: (i) a new way to examine overlap data; (ii) a basis on which to compare overlap data and probe data using the same systems and standards; and (iii) a unique and useful way to uniformly integrate overlap data with probe data. PMID- 11395426 TI - Aligning gene expression time series with time warping algorithms. AB - motivation: Increasingly, biological processes are being studied through time series of RNA expression data collected for large numbers of genes. Because common processes may unfold at varying rates in different experiments or individuals, methods are needed that will allow corresponding expression states in different time series to be mapped to one another. RESULTS: We present implementations of time warping algorithms applicable to RNA and protein expression data and demonstrate their application to published yeast RNA expression time series. Programs executing two warping algorithms are described, a simple warping algorithm and an interpolative algorithm, along with programs that generate graphics that visually present alignment information. We show time warping to be superior to simple clustering at mapping corresponding time states. We document the impact of statistical measurement noise and sample size on the quality of time alignments, and present issues related to statistical assessment of alignment quality through alignment scores. We also discuss directions for algorithm improvement including development of multiple time series alignments and possible applications to causality searches and non-temporal processes ('concentration warping'). PMID- 11395427 TI - A Bayesian framework for the analysis of microarray expression data: regularized t -test and statistical inferences of gene changes. AB - MOTIVATION: DNA microarrays are now capable of providing genome-wide patterns of gene expression across many different conditions. The first level of analysis of these patterns requires determining whether observed differences in expression are significant or not. Current methods are unsatisfactory due to the lack of a systematic framework that can accommodate noise, variability, and low replication often typical of microarray data. RESULTS: We develop a Bayesian probabilistic framework for microarray data analysis. At the simplest level, we model log expression values by independent normal distributions, parameterized by corresponding means and variances with hierarchical prior distributions. We derive point estimates for both parameters and hyperparameters, and regularized expressions for the variance of each gene by combining the empirical variance with a local background variance associated with neighboring genes. An additional hyperparameter, inversely related to the number of empirical observations, determines the strength of the background variance. Simulations show that these point estimates, combined with a t -test, provide a systematic inference approach that compares favorably with simple t -test or fold methods, and partly compensate for the lack of replication. PMID- 11395428 TI - Missing value estimation methods for DNA microarrays. AB - MOTIVATION: Gene expression microarray experiments can generate data sets with multiple missing expression values. Unfortunately, many algorithms for gene expression analysis require a complete matrix of gene array values as input. For example, methods such as hierarchical clustering and K-means clustering are not robust to missing data, and may lose effectiveness even with a few missing values. Methods for imputing missing data are needed, therefore, to minimize the effect of incomplete data sets on analyses, and to increase the range of data sets to which these algorithms can be applied. In this report, we investigate automated methods for estimating missing data. RESULTS: We present a comparative study of several methods for the estimation of missing values in gene microarray data. We implemented and evaluated three methods: a Singular Value Decomposition (SVD) based method (SVDimpute), weighted K-nearest neighbors (KNNimpute), and row average. We evaluated the methods using a variety of parameter settings and over different real data sets, and assessed the robustness of the imputation methods to the amount of missing data over the range of 1--20% missing values. We show that KNNimpute appears to provide a more robust and sensitive method for missing value estimation than SVDimpute, and both SVDimpute and KNNimpute surpass the commonly used row average method (as well as filling missing values with zeros). We report results of the comparative experiments and provide recommendations and tools for accurate estimation of missing microarray data under a variety of conditions. PMID- 11395429 TI - Database verification studies of SWISS-PROT and GenBank. AB - PROBLEM STATEMENT: We have studied the relationships among SWISS-PROT, TrEMBL, and GenBank with two goals. First is to determine whether users can reliably identify those proteins in SWISS-PROT whose functions were determined experimentally, as opposed to proteins whose functions were predicted computationally. If this information was present in reasonable quantities, it would allow researchers to decrease the propagation of incorrect function predictions during sequence annotation, and to assemble training sets for developing the next generation of sequence-analysis algorithms. Second is to assess the consistency between translated GenBank sequences and sequences in SWISS-PROT and TrEMBL. RESULTS: (1) Contrary to claims by the SWISS-PROT authors, we conclude that SWISS-PROT does not identify a significant number of experimentally characterized proteins. (2) SWISS-PROT is more incomplete than we expected in that version 38.0 from July 1999 lacks many proteins from the full genomes of important organisms that were sequenced years earlier. (3) Even if we combine SWISS-PROT and TrEMBL, some sequences from the full genomes are missing from the combined dataset. (4) In many cases, translated GenBank genes do not exactly match the corresponding SWISS-PROT sequences, for reasons that include missing or removed methionines, differing translation start positions, individual amino-acid differences, and inclusion of sequence data from multiple sequencing projects. For example, results show that for Escherichia coli, 80.6% of the proteins in the GenBank entry for the complete genome have identical sequence matches with SWISS-PROT/TrEMBL sequences, 13.4% have exact substring matches, and matches for 4.1% can be found using BLAST search; the remaining 2.0% of E.coli protein sequences (most of which are ORFs) have no clear matches to SWISS PROT/TrEMBL. Although many of these differences can be explained by the complexity of the DB, and by the curation processes used to create it, the scale of the differences is notable. PMID- 11395431 TI - TreeGeneBrowser: phylogenetic data mining of gene sequences from public databases. AB - MOTIVATION: Sequence databases represent an enormous resource of phylogenetic information, but there is a lack of tools for accessing that information in order to assess the amount of evolutionary information in these databases that may be suitable for phylogenetic reconstruction and for identifying areas of the taxonomy that are under-represented for specific gene sequences. RESULTS: We have developed TreeGeneBrowser which allows inspection and evaluation of gene sequence data for phylogenetic reconstruction. This program improves the efficiency of identification of genes that may be useful for particular phylogenetic studies and identifies taxa and taxonomic branches that are under-represented in sequence databases. PMID- 11395432 TI - SCORE: predicting the core of protein models. AB - MOTIVATION: The prediction of the regions of homology models that can be 'restrained by' or 'copied from' the basis structures is a vital step in correct model generation, because these regions are the models most accurate part. However, there is no ideal method for the identification of their limits. In most algorithms their length depends on the number of family members and definitions of secondary structure. RESULTS: The algorithm SCORE steps away from the conventional definitions of the core to identify from large numbers of basis structures those regions that can be considered structurally related to a target sequence. The use of phi, psi constraints to accurately pinpoint the regions that are conserved across a family and environmentally constrained substitution tables to extend these regions allows SCORE to rapidly (generally in under 1 s, an order of magnitude faster than methods such as MODELLER) identify and build the core of homology models from the alignments of the target sequence to the basis structures. The SCORE algorithm was used to build 114 model cores. In only two cases was the core size less than 50% of the structure and all the cores built had an RMSD of 3.7 A or less to the target structure. PMID- 11395433 TI - Prediction of quaternary structure from primary structure. AB - MOTIVATION: The 'sequence implies conformation' principle has been the motivation for the construction of numerous systems of secondary and tertiary structure prediction. Computational experiments have shown that this principle can now be extended to quaternary structure prediction. This work appears to be the first effort to predict quaternary structure properties from sequence. RESULTS: The software developed to conduct these experiments was the Quaternary Structure Explorer (QSE). Successful rule-based classifiers have been found that can discriminate between the primary sequences of homodimers and non-homodimers. PMID- 11395434 TI - Compositional symmetries in complete genomes. PMID- 11395435 TI - Identification of novel families of membrane proteins from the model plant Arabidopsis thaliana. AB - MOTIVATION: The completion of the Arabidopsis genome offers the first opportunity to analyze all of the membrane protein sequences of a plant. The majority of integral membrane proteins including transporters, channels, and pumps contain hydrophobic alpha-helices and can be selected based on TransMembrane Spanning (TMS) domain prediction. By clustering the predicted membrane proteins based on sequence, it is possible to sort the membrane proteins into families of known function, based on experimental evidence or homology, or unknown function. This provides a way to identify target sequences for future functional analysis. RESULTS: An automated approach was used to select potential membrane protein sequences from the set of all predicted proteins and cluster the sequences into related families. The recently completed sequence of Arabidopsis thaliana, a model plant, was analyzed. Of the 25,470 predicted protein sequences 4589 (18%) were identified as containing two or more membrane spanning domains. The membrane protein sequences clustered into 628 distinct families containing 3208 sequences. Of these, 211 families (1764 sequences) either contained proteins of known function or showed homology to proteins of known function in other species. However, 417 families (1444 sequences) contained only sequences with no known function and no homology to proteins of known function. In addition, 1381 sequences did not cluster with any family and no function could be assigned to 1337 of these. PMID- 11395436 TI - MAPS: a microarray project system for gene expression experiment information and data validation. AB - SUMMARY: MAPS is a MicroArray Project System for management and interpretation of microarray gene expression experiment information and data. Microarray project information is organized to track experiments and results that are: (1) validated by performing analysis on stored replicate gene expression data; and (2) queried according to the biological classifications of genes deposited on microarray chips. PMID- 11395437 TI - SVDMAN--singular value decomposition analysis of microarray data. AB - SUMMARY: We have developed two novel methods for Singular Value Decomposition analysis (SVD) of microarray data. The first is a threshold-based method for obtaining gene groups, and the second is a method for obtaining a measure of confidence in SVD analysis. Gene groups are obtained by identifying elements of the left singular vectors, or gene coefficient vectors, that are greater in magnitude than the threshold W N(-1/2), where N is the number of genes, and W is a weight factor whose default value is 3. The groups are non-exclusive and may contain genes of opposite (i.e. inversely correlated) regulatory response. The confidence measure is obtained by systematically deleting assays from the data set, interpolating the SVD of the reduced data set to reconstruct the missing assay, and calculating the Pearson correlation between the reconstructed assay and the original data. This confidence measure is applicable when each experimental assay corresponds to a value of parameter that can be interpolated, such as time, dose or concentration. Algorithms for the grouping method and the confidence measure are available in a software application called SVD Microarray ANalysis (SVDMAN). In addition to calculating the SVD for generic analysis, SVDMAN provides a new means for using microarray data to develop hypotheses for gene associations and provides a measure of confidence in the hypotheses, thus extending current SVD research in the area of global gene expression analysis. PMID- 11395438 TI - AMADA: analysis of microarray data. AB - SUMMARY: AMADA is a Windows program for identifying co-expressed genes from microarray data. It performs data transformation, principal component analysis, a variety of cluster analyses and extensive graphic functions for visualizing expression profiles. PMID- 11395439 TI - The HIB database of annotated UniGene clusters. AB - SUMMARY: The HumanInfoBase (HIB) is a database of putative human gene transcripts. UniGene clusters are assembled, and the resulting consensus sequences are submitted to the PEDANT software system (Frishman,D., Albermann,K., Hani,J., Heumann,K., Metanomski,A., Zollner,A. and Mewes,H.-W., 2001, Bioinformatics, 17, 44--57) for fully automatic sequence analysis and annotation. Predicted transcripts are classified using a variety of functional and structural categories, and hyperlinks to various databases are provided for additional information. A WWW-based graphical user interface represents the assembly process as well as functionally important sites in the putative transcripts. PMID- 11395440 TI - SOAP, cleaning multiple alignments from unstable blocks. AB - SUMMARY: SOAP is a stand-alone, multi-platform program to test the stability of a multiple alignment of molecular sequences. PMID- 11395441 TI - STOCHSIM: modelling of stochastic biomolecular processes. AB - SUMMARY: STOCHSIM is a stochastic simulator for chemical reactions. Molecules are represented as individual software objects that react according to probabilities derived from concentrations and rate constants. Version 1.2 of STOCHSIM provides a novel cross-platform graphical interface written in Perl/Tk. A simple two dimensional spatial structure has also been implemented, in which nearest neighbour interactions of molecules in a 2-D lattice can be simulated. PMID- 11395442 TI - SeWeR: a customizable and integrated dynamic HTML interface to bioinformatics services. AB - SUMMARY: Sequence analysis using Web Resources (SeWeR) is an integrated, Dynamic HTML (DHTML) interface to commonly used bioinformatics services available on the World Wide Web. It is highly customizable, extendable, platform neutral, completely server-independent and can be hosted as a web page as well as being used as stand-alone software running within a web browser. PMID- 11395443 TI - Molecular physiology of sugar catabolism in Lactococcus lactis IL1403. AB - The metabolic characteristics of Lactococcus lactis IL1403 were examined on two different growth media with respect to the physiological response to two sugars, glucose and galactose. Analysis of specific metabolic rates indicated that despite significant variations in the rates of both growth and sugar consumption, homolactic fermentation was maintained for all cultures due to the low concentration of either pyruvate-formate lyase or alcohol dehydrogenase. When the ionophore monensin was added to the medium, flux through glycolysis was not increased, suggesting a catabolic flux limitation, which, with the low intracellular concentrations of glycolytic intermediates and high in vivo glycolytic enzyme capacities, may be at the level of sugar transport. To assess transcription, a novel DNA macroarray technology employed RNA labeled in vitro with digoxigenin and detection of hybrids with an alkaline phosphatase antidigoxigenin conjugate. This method showed that several genes of glycolysis were expressed to higher levels on glucose and that the genes of the mixed-acid pathway were expressed to higher levels on galactose. When rates of enzyme synthesis are compared to transcript concentrations, it can be deduced that some translational regulation occurs with threefold-higher translational efficiency in cells grown on glucose. PMID- 11395444 TI - Analysis of the PilQ secretin from Neisseria meningitidis by transmission electron microscopy reveals a dodecameric quaternary structure. AB - PilQ is a member of the secretin family of outer membrane proteins and is specifically involved in secretion of type IV pili in Neisseria meningitidis, Neisseria gonorrhoeae, and Pseudomonas aeruginosa. The quaternary structure of PilQ from N. meningitidis was analyzed by transmission electron microscopy by using a negative stain. Single particle averaging was carried out with a total data set of 650 individual particles, which produced a projection map generated from 296 particles at an estimated resolution of 2.6 nm. Oligomeric PilQ adopts a donut-like structure with an external ring that is 16.5 nm in diameter surrounding a central cavity that is 6.5 nm in diameter. Self-rotation and power spectrum analysis demonstrated the presence of 12-fold rotational symmetry, showing that PilQ is organized as a ring of 12 identical subunits. A model of the type IV meningococcal pilus fiber, based on the X-ray crystal structure of the N. gonorrhoeae pilin subunit, fitted neatly into the cavity, demonstrating how PilQ could serve as a channel for the growing pilus fiber. PMID- 11395445 TI - Autoregulation of the dnaA-dnaN operon and effects of DnaA protein levels on replication initiation in Bacillus subtilis. AB - In Escherichia coli, the DnaA protein level appears to play a pivotal role in determining the timing of replication initiation. To examine the effects on replication initiation in B. subtilis, we constructed a strain in which a copy of the dnaA gene was integrated at the purA locus on the chromosome under the control of an isopropyl-beta-D-thiogalactopyranoside (IPTG)-inducible promoter. However, increasing the DnaA level resulted in cell elongation and inhibition of cell growth by induction of the SOS response. Transcription of the native dnaA dnaN operon was greatly reduced at high DnaA levels, but it was increased in a dnaA-null mutant, indicating autoregulation of the operon by DnaA. When a copy of the dnaN gene was added downstream of the additional dnaA gene at purA, the cells grew at high DnaA levels, suggesting that depletion of DnaN (beta subunit of DNA polymerase III) within the cell by repression of the native dnaA-dnaN operon at high DnaA levels was the cause of the SOS induction. Flow cytometry of the cells revealed that the cell mass at initiation of replication increased at a lower DnaA level and decreased at DnaA levels higher than those of the wild type. Proper timing of replication initiation was observed at DnaA levels nearly comparable to the wild-type level. These results suggest that if the DnaA level increases with progression of the replication cycle, it could act as a rate limiting factor of replication initiation in B. subtilis. PMID- 11395446 TI - Identification of a dedicated recycling pathway for anhydro-N-acetylmuramic acid and N-acetylglucosamine derived from Escherichia coli cell wall murein. AB - Turnover and recycling of the cell wall murein represent a major metabolic pathway of Escherichia coli. It is known that E. coli efficiently reuses, i.e., recycles, its murein tripeptide, L-alanyl-gamma-D-glutamyl-meso-diaminopimelate, to form new murein. However, the question of whether the cells also recycle the amino sugar moieties of cell wall murein has remained unanswered. It is demonstrated here that E. coli recycles the N-acetylglucosamine present in cell wall murein degradation products for de novo murein and lipopolysaccharide synthesis. Furthermore, E. coli also recycles the anhydro-N-acetylmuramic acid moiety by first converting it into N-acetylglucosamine. Based on the results obtained by studying mutants unable to recycle amino sugars, the pathway for recycling is revealed. PMID- 11395447 TI - Increased expression of Escherichia coli polynucleotide phosphorylase at low temperatures is linked to a decrease in the efficiency of autocontrol. AB - Polynucleotide phosphorylase (PNPase) synthesis is translationally autocontrolled via an RNase III-dependent mechanism, which results in a tight correlation between protein level and messenger stability. In cells grown at 18 degrees C, the amount of PNPase is twice that found in cells grown at 30 degrees C. To investigate whether this effect was transcriptional or posttranscriptional, the expression of a set of pnp-lacZ transcriptional and translational fusions was analyzed in cells grown at different temperatures. In the absence of PNPase, there was no increase in pnp-lacZ expression, indicating that the increase in pnp expression occurs at a posttranscriptional level. Other experiments clearly show that increased pnp expression at low temperature is only observed under conditions in which the autocontrol mechanism of PNPase is functional. At low temperature, the destabilizing effect of PNPase on its own mRNA is less efficient, leading to a decrease in repression and an increase in the expression level. PMID- 11395448 TI - Activities of virE1 and the VirE1 secretion chaperone in export of the multifunctional VirE2 effector via an Agrobacterium type IV secretion pathway. AB - Agrobacterium tumefaciens uses a type IV secretion system to deliver oncogenic nucleoprotein particles and effector proteins, such as the multifunctional VirE2 protein, to plant cells. In this study, we examined the function of virE1 and its product, the VirE1 secretion chaperone, in mediating VirE2 export. A nonpolar virE1 null mutant accumulated low levels of VirE2, and trans expression of virE1 in this mutant only partially restored VirE2 abundance. Deletion of virE1 did not affect transcription but decreased translation of virE2, as shown by analysis of lacZ transcriptional and translational fusions. VirE2 was stable for a prolonged period, more than 6 h, when it was expressed in cis with virE1, and it exhibited half-lives of about 2 h when it was expressed in trans with virE1 and less than 10 min when it was expressed in the absence of virE1, as shown by pulse-chase experiments. VirE1 stabilized VirE2 via an interaction with a domain near the N terminus of VirE2, as shown by analyses of VirE2 truncation and insertion mutants synthesized in A. tumefaciens. VirE1 self-association was demonstrated by using bacteriophage lambda cI repressor fusion and pull-down assays, and evidence of VirE1 homomultimerization in vivo was obtained by native polyacrylamide gel electrophoresis and gel filtration chromatography. A putative VirE1-VirE2 complex with a molecular mass of about 70 to 80 kDa was detected by gel filtration chromatography of extracts from wild-type cells, whereas higher-order VirE2 complexes or aggregates were detected in extracts from a virE1 mutant. Taken together, our findings show that virE1 contributes in several ways to VirE2 export:(i) virE1 regulates efficient virE2 translation in the context of expression from the native P(virE) promoter; (ii) the VirE1 secretion chaperone stabilizes VirE2, most probably via an interaction with an N-terminal domain; and (iii) VirE1 forms a VirE1-VirE2 complex with a predicted 2:1 stoichiometry that inhibits assembly of higher-order VirE2 complexes or aggregates. PMID- 11395449 TI - In vitro processing of the 16S rRNA of the thermophilic archaeon Sulfolobus solfataricus. AB - In this paper we have analyzed the processing in vitro of the 16S rRNA of the thermophilic archaeon Sulfolobus solfataricus, using pre-rRNA substrates transcribed in vitro and different protein preparations as the source of processing enzymes. We show that the 5' external transcribed spacer of the S. solfataricus pre-rRNA transcript contains a target site for a specific endonuclease, which recognizes a conserved sequence also existing in the early A0 and 0 processing sites of Saccharomyces cerevisiae and vertebrates. This site is present in other members of the kingdom Crenarchaeota but apparently not in the Euryarchaeota. Furthermore, S. solfataricus pre-16S RNA is processed within the double-helical stem formed by the inverted repeats flanking the 16S RNA sequence, in correspondence with a bulge-helix-bulge motif. The endonuclease responsible for this cleavage is present in both the Crenarchaeota and the Euryarchaeota. The processing pattern remained the same when the substrate was a 30S ribonucleoprotein particle instead of the naked RNA. Maturation of either the 5' or the 3' end of the 16S RNA molecule was not observed, suggesting either that maturation requires conditions not easily reproducible in vitro or that the responsible endonucleases are scarcely represented in cell extracts. PMID- 11395450 TI - Promoters of the CATG-specific methyltransferase gene hpyIM differ between iceA1 and iceA2 Helicobacter pylori strains. AB - Helicobacter pylori strains can be divided into two groups, based on the presence of two unrelated genes, iceA1 and iceA2, that occupy the same genomic locus. hpyIM, located immediately downstream of either gene, encodes a functional CATG specific methyltransferase. Despite the strong conservation of the hpyIM open reading frame (ORF) among all H. pylori strains, the sequences upstream of the ORF in iceA1 and iceA2 strains are substantially different. To explore the roles of these upstream sequences in hpyIM regulation, promoter analysis of hpyIM was performed. Both deletion mutation and primer extension analyses demonstrate that the hpyIM promoters differ between H. pylori strains 60190 (iceA1) and J188 (iceA2). In strain 60190, hpyIM has two promoters, P(a) or P(I), which may function independently, whereas only one hpyIM promoter, P(c), was found in strain J188. The XylE assay showed that the hpyIM transcription level was much higher in strain 60190 than in strain J188, indicating that regulation of hpyIM transcription differs between the H. pylori iceA1 strain (60190) and iceA2 strains (J188). Since the iceA1 and iceA2 sequences are highly conserved within iceA1 or iceA2 strains, we conclude that promoters of the CATG-specific methylase gene hpyIM differ between iceA1 and iceA2 strains, which leads to differences in regulation of hpyIM transcription. PMID- 11395451 TI - SsrA-mediated tagging in Bacillus subtilis. AB - A general mechanism in bacteria to rescue stalled ribosomes involves a stable RNA encoded by the ssrA gene. This RNA, termed tmRNA, encodes a proteolytic peptide tag which is cotranslationally added to truncated polypeptides, thereby targeting them for rapid proteolysis. To study this ssrA-mediated mechanism in Bacillus subtilis, a bipartite detection system was constructed that was composed of the HrcA transcriptional repressor and the bgaB reporter gene coding for a heat stable beta-galactosidase fused to an HrcA-controlled promoter. After the predicted proteolysis tag was fused to HrcA, the reporter beta-galactosidase was expressed constitutively at a high level due to the instability of the tagged HrcA. Replacement of the two C-terminal alanine residues of the tag by aspartate rendered the repressor stable. Replacement of the hrcA stop codon by a transcriptional terminator sequence rendered the protein unstable; this was caused by trans translational addition of the proteolytic tag. Inactivating the B. subtilis ssrA or smpB (yvaI) gene prevented the trans translational tagging reaction. Various protease-deficient strains of B. subtilis were tested for proteolysis of tagged HrcA. HrcA remained stable only in clpX or clpP knockouts, which suggests that this ATP-dependent protease is primarily responsible for the degradation of SsrA-tagged proteins in B. subtilis. PMID- 11395452 TI - Genome-wide transcriptional profiling of the Escherichia coli responses to superoxide stress and sodium salicylate. AB - Escherichia coli responds to oxidative stress by activating sets of coregulated genes that help the cell to maintain homeostasis. Identified previously by genetic and biochemical approaches, the soxRS system mediates the induction of 18 of these redox-inducible genes (including the soxS gene itself). An overlapping set of genes is activated by an assortment of structurally unrelated molecules with antibiotic activities; many genes in this response are controlled by the marRAB system. The activation of either the soxRS or the marRAB system results in enhanced resistance to both superoxide-generating agents and multiple antibiotics. In order to probe the extent of these regulatory networks, we have measured whole-genome transcriptional profiles of the E. coli response to the superoxide-generating agent paraquat (PQ), an inducer of the soxRS system, and to the weak acid salt sodium salicylate (NaSal), an inducer of the marRA system. A total of 112 genes was modulated in response to PQ, while 134 genes were modulated in response to NaSal. We have also obtained transcriptional profiles of the SoxS and MarA regulons in the absence of global stress, in order to establish the regulatory hierarchies within the global responses. Several previously unrelated genes were shown to be under SoxS or MarA control. The genetic responses to both environmental insults revealed several common themes, including the activation of genes coding for functions that replenish reducing potential; regulate iron transport and storage; and participate in sugar and amino acid transport, detoxification, protein modification, osmotic protection, and peptidoglycan synthesis. A large number of PQ- and NaSal-responsive genes have no known function, suggesting that many adaptive metabolic changes that ensue after stress remain uncharacterized. PMID- 11395454 TI - Activation from a distance: roles of Lrp and integration host factor in transcriptional activation of gltBDF. AB - The leucine-responsive regulatory protein (Lrp) binds to three sites centered 252, 216, and 152 bp upstream of the transcription start site of the Escherichia coli glutamate synthase operon (gltBDF) and activates transcription. Activators of sigma(70)-dependent promoters usually bind closer to the -35 hexamer of the core promoter sequence. To study the mechanism by which Lrp-dependent activation occurs over this relatively large distance, the gltBDF upstream region was sequentially replaced with corresponding portions from the well-characterized sigma(70)-dependent promoter lacZYAp. The glt-lac promoter hybrids were placed upstream of lacZ, allowing transcriptional activity to be monitored via beta galactosidase assays. Even replacing all gltBDF sequences downstream of and including the -35 hexamer did not eliminate Lrp-dependent activation of transcription. When a 91-bp region between the -35 hexamer and the proximal Lrp binding site (-48 to -128) was replaced with heterologous DNA of the same length, transcription was reduced nearly 40-fold. Based on the presence of a consensus binding sequence, this region seemed likely to be a binding site for integration host factor (IHF). Experiments to study the effects of a himD mutant on expression of a gltB::lacZ transcriptional fusion, gel mobility shift analyses, and DNA footprinting assays were used to confirm the direct participation of IHF in gltBDF promoter regulation. Based on these results, we suggest that IHF plays a crucial architectural role, bringing the distant Lrp complex in close proximity to the promoter-bound RNA polymerase. PMID- 11395453 TI - Signaling system in Porphyromonas gingivalis based on a LuxS protein. AB - The luxS gene of quorum-sensing Vibrio harveyi is required for type 2 autoinducer production. We identified a Porphyromonas gingivalis open reading frame encoding a predicted peptide of 161 aa that shares 29% identity with the amino acid sequence of the LuxS protein of V. harveyi. Conditioned medium from a late-log phase P. gingivalis culture induced the luciferase operon of V. harveyi, but that from a luxS insertional mutant did not. In P. gingivalis, the expression of luxS mRNA was environmentally controlled and varied according to the cell density and the osmolarity of the culture medium. In addition, differential display PCR showed that the inactivation of P. gingivalis luxS resulted in up-regulation of a hemin acquisition protein and an arginine-specific protease and reduced expression of a hemin-regulated protein, a TonB homologue, and an excinuclease. The data suggest that the luxS gene in P. gingivalis may function to control the expression of genes involved in the acquisition of hemin. PMID- 11395455 TI - The Agrobacterium tumefaciens rnd homolog is required for TraR-mediated quorum dependent activation of Ti plasmid tra gene expression. AB - Conjugal transfer of Agrobacterium tumefaciens Ti plasmids is regulated by quorum sensing via TraR and its cognate autoinducer, N-(3-oxo-octanoyl)-L-homoserine lactone. We isolated four Tn5-induced mutants of A. tumefaciens C58 deficient in TraR-mediated activation of tra genes on pTiC58DeltaaccR. These mutations also affected the growth of the bacterium but had no detectable influence on the expression of two tester gene systems that are not regulated by quorum sensing. In all four mutants Tn5 was inserted in a chromosomal open reading frame (ORF) coding for a product showing high similarity to RNase D, coded for by rnd of Escherichia coli, an RNase known to be involved in tRNA processing. The wild-type allele of the rnd homolog cloned from C58 restored the two phenotypes to each mutant. Several ORFs, including a homolog of cya2, surround A. tumefaciens rnd, but none of these genes exerted a detectable effect on the expression of the tra reporter. In the mutant, traR was expressed from the Ti plasmid at a level about twofold lower than that in NT1. The expression of tra, but not the growth rate, was partially restored by increasing the copy number of traR or by disrupting traM, a Ti plasmid gene coding for an antiactivator specific for TraR. The mutation in rnd also slightly reduced expression of two tested vir genes but had no detectable effect on tumor induction by this mutant. Our data suggest that the defect in tra gene induction in the mutants results from lowered levels of TraR. In turn, production of sufficient amounts of TraR apparently is sensitive to a cellular function requiring RNase D. PMID- 11395456 TI - Intra- and interspecies signaling between Streptococcus salivarius and Streptococcus pyogenes mediated by SalA and SalA1 lantibiotic peptides. AB - Streptococcus salivarius 20P3 produces a 22-amino-acid residue lantibiotic, designated salivaricin A (SalA), that inhibits the growth of a range of streptococci, including all strains of Streptococcus pyogenes. Lantibiotic production is associated with the sal genetic locus comprising salA, the lantibiotic structural gene; salBCTX genes encoding peptide modification and export machinery proteins; and salYKR genes encoding a putative immunity protein and two-component sensor-regulator system. Insertional inactivation of salB in S. salivarius 20P3 resulted in abrogation of SalA peptide production, of immunity to SalA, and of salA transcription. Addition of exogenous SalA peptide to salB mutant cultures induced dose-dependent expression of salA mRNA (0.2 kb), demonstrating that SalA production was normally autoregulated. Inactivation of salR encoding the response regulator of the SalKR two-component system led to reduced production of, and immunity to, SalA. The sal genetic locus was also present in S. pyogenes SF370 (M type 1), but because of a deletion across the salBCT genes, the corresponding lantibiotic peptide, designated SalA1, was not produced. However, in S. pyogenes T11 (M type 4) the sal locus gene complement was apparently complete, and active SalA1 peptide was synthesized. Exogenously added SalA1 peptide from S. pyogenes T11 induced salA1 transcription in S. pyogenes SF370 and in an isogenic S. pyogenes T11 salB mutant and salA transcription in S. salivarius 20P3 salB. Thus, SalA and SalA1 are examples of streptococcal lantibiotics whose production is autoregulated. These peptides act as intra- and interspecies signaling molecules, modulating lantibiotic production and possibly influencing streptococcal population ecology in the oral cavity. PMID- 11395458 TI - Genetic organization of plasmid ColJs, encoding colicin Js activity, immunity, and release genes. AB - The 5.2-kb ColJs plasmid of a colicinogenic strain of Shigella sonnei (colicin type 7) was isolated and sequenced. pColJs was partly homologous to pColE1 and to pesticin-encoding plasmid pPCP1, mainly in the rep, mob, and cer regions. A 1.2 kb unique region of pColJs showed significantly different G+C content (34%) compared to the rest of pColJs (53%). Within the unique region, seven open reading frames (ORFs) were identified. ORF94 was shown to code for colicin Js activity (cja), a 94-amino-acid polypeptide (molecular mass, 10.4 kDa); ORF129 (cji) was shown to code for the 129-amino-acid colicin Js immunity protein (molecular mass, 14.3 kDa); and ORF65 was shown to be involved in colicin Js release by producer bacteria (cjl) coding for a 65-amino-acid polypeptide (molecular mass, 7.5 kDa). In contrast to the gene order in other colicin operons, the cjl gene was found upstream from cja. Moreover, the promoter upstream from cjl was similar to promoters described upstream from several colicin activity genes. The cji gene was found to be located downstream from cja with a transcription polarity opposite to that of the cjl and cja genes. The cja, cji, and cjl genes were not similar to other known colicin genes. Colicin Js was purified as an inactive fusion protein with an N-terminal histidine tag. Activity of the purified fusion form of colicin Js was restored after cleavage of the amino acids fused to the colicin Js N terminus. PMID- 11395457 TI - Activity of the Kluyveromyces lactis Pdr5 multidrug transporter is modulated by the Sit4 protein phosphatase. AB - A possible role for posttranslational modifications in regulating the activity of ATP-binding cassette (ABC) transporters has not been well established. In this study, the drug efflux ABC transporter gene KlPDR5 was isolated from the budding yeast Kluyveromyces lactis, and it was found that the encoded KlPdr5 drug pump is posttranslationally regulated by the type 2A-related Ser/Thr protein phosphatase, Sit4p. The KlPdr5 transporter is a protein of 1,525 amino acids sharing 63.8% sequence identity with its Saccharomyces cerevisiae counterpart, ScPdr5p. Overexpression of the KlPDR5 gene confers resistance to oligomycin, antimycin, econazole, and ketoconazole, whereas cells with a disrupted allele of KlPDR5 are hypersensitive to the drugs and have a decreased capacity to carry out efflux of the anionic fluorescent dye rhodamine 123. It was found that a chromosomal disruption of KlPDR5 abolishes the drug-resistant phenotype associated with sit4 mutations and that a synergistic hyperresistance to the drugs can be created by overexpressing KlPDR5 in sit4 mutants. These data strongly indicate that the multidrug-resistant phenotype of sit4 mutants is mediated by negatively modulating the activity of KlPdr5p. As the transcriptional level of KlPDR5 and the steady-state level of KlPdr5p are not significantly affected by mutations in SIT4, the regulation by Sit4p appears to be a posttranslational process. PMID- 11395459 TI - The iron- and temperature-regulated cjrBC genes of Shigella and enteroinvasive Escherichia coli strains code for colicin Js uptake. AB - A cosmid library of DNA from colicin Js-sensitive enteroinvasive Escherichia coli (EIEC) strain O164 was made in colicin Js-resistant strain E. coli VCS257, and colicin Js-sensitive clones were identified. Sensitivity to colicin Js was associated with the carriage of a three-gene operon upstream of and partially overlapping senB. The open reading frames were designated cjrABC (for colicin Js receptor), coding for proteins of 291, 258, and 753 amino acids, respectively. Tn7 insertions in any of them led to complete resistance to colicin Js. A near consensus Fur box was found upstream of cjrA, suggesting regulation of the cjr operon by iron levels. CjrA protein was homologous to iron-regulated Pseudomonas aeruginosa protein PhuW, whose function is unknown; CjrB was homologous to the TonB protein from Pseudomonas putida; and CjrC was homologous to a putative outer membrane siderophore receptor from Campylobacter jejuni. Cloning experiments showed that the cjrB and cjrC genes are sufficient for colicin Js sensitivity. Uptake of colicin Js into sensitive bacteria was dependent on the ExbB protein but not on the E. coli K-12 TonB and TolA, -B, and -Q proteins. Sensitivity to colicin Js is positively regulated by temperature via the VirB protein and negatively controlled by the iron source through the Fur protein. Among EIEC strains, two types of colicin Js-sensitive phenotypes were identified that differed in sensitivity to colicin Js by 1 order of magnitude. The difference in sensitivity to colicin Js is not due to differences between the sequences of the CjrB and CjrC proteins. PMID- 11395460 TI - Three efflux pumps are required to provide efficient tolerance to toluene in Pseudomonas putida DOT-T1E. AB - In Pseudomonas putida DOT-T1E multidrug efflux pumps of the resistance-nodulation division family make a major contribution to solvent resistance. Two pumps have been identified: TtgABC, expressed constitutively, and TtgDEF, induced by aromatic hydrocarbons. A double mutant lacking both efflux pumps was able to survive a sudden toluene shock if and only if preinduced with small amounts of toluene supplied via the gas phase. In this article we report the identification and characterization in this strain of a third efflux pump, named TtgGHI. The ttgGHI genes form an operon that is expressed constitutively at high levels from a single promoter. In the presence of toluene the operon is expressed at an even higher level from two promoters, the constitutive one and a previously unreported one that is inducible and that partially overlaps the constitutive promoter. By site-directed mutagenesis we constructed a single ttgH mutant which was shown to be unable to survive sudden 0.3% (vol/vol) toluene shocks regardless of the preculture conditions. The mutation was transferred to single and double mutants to construct mutant strains in which two or all three pumps are knocked out. Survival analysis of induced and noninduced cells revealed that the TtgABC and TtgGHI pumps extruded toluene, styrene, m-xylene, ethylbenzene, and propylbenzene, whereas the TtgDEF pump removed only toluene and styrene. The triple mutant was hypersensitive to toluene, as shown by its inability to grow with toluene supplied via the vapor phase. PMID- 11395461 TI - NadN and e (P4) are essential for utilization of NAD and nicotinamide mononucleotide but not nicotinamide riboside in Haemophilus influenzae. AB - Haemophilus influenzae has an absolute requirement for NAD (factor V) because it lacks almost all the biosynthetic enzymes necessary for the de novo synthesis of that cofactor. Factor V can be provided as either nicotinamide adenosine dinucleotide (NAD), nicotinamide mononucleotide (NMN), or nicotinamide riboside (NR) in vitro, but little is known about the source or the mechanism of uptake of these substrates in vivo. As shown by us earlier, at least two gene products are involved in the uptake of NAD, the outer membrane lipoprotein e (P4), which has phosphatase activity and is encoded by hel, and a periplasmic NAD nucleotidase, encoded by nadN. It has also been observed that the latter gene product is essential for H. influenzae growth on media supplemented with NAD. In this report, we describe the functions and substrates of these two proteins as they act together in an NAD utilization pathway. Data are provided which indicate that NadN harbors not only NAD pyrophosphatase but also NMN 5'-nucleotidase activity. The e (P4) protein is also shown to have NMN 5'-nucleotidase activity, recognizing NMN as a substrate and releasing NR as its product. Insertion mutants of nadN or deletion and site-directed mutants of hel had attenuated growth and a reduced uptake phenotype when NMN served as substrate. A hel and nadN double mutant was only able to grow in the presence of NR, whereas no uptake of NMN was observed. PMID- 11395462 TI - Localization of a germinant receptor protein (GerBA) to the inner membrane of Bacillus subtilis spores. AB - Dormant Bacillus subtilis spores germinate in response to specific nutrients called germinants, which are recognized by multisubunit receptor complexes encoded by members of the gerA family of operons, of which the gerB operon is a member. The germinant receptors are expected to be membrane associated, but there is some debate about whether they are located in the inner or outer spore membrane. In this study we have used Western blot analysis to determine the precise location of GerBA, a gerB-encoded receptor protein, in various spore fractions. GerBA was not extracted from spores by a decoating treatment that removes the coat and outer membrane but was present in lysates from decoated spores and in the insoluble fraction (termed P100) from such lysates that contained inner-membrane vesicles. GerBA was also solubilized from the P100 fraction with detergent but not with high salt. These findings suggest that GerBA is an integral membrane protein located in the spore's inner membrane. Consistent with this idea, GerBA was present in the cell membrane of the outgrowing spore, a membrane that is derived from the dormant spore's inner membrane. Based on these observations we propose that GerBA and probably the entire GerB germinant receptor are located in the inner membrane of the dormant spore. We also estimated that there are only 24 to 40 molecules of GerBA per spore, a number that is consistent with the previously reported low level of gerB operon expression and with the putative receptor function of the proteins encoded by the gerB operon. PMID- 11395463 TI - Cell wall core galactofuran synthesis is essential for growth of mycobacteria. AB - The mycobacterial cell wall core consists of an outer lipid (mycolic acid) layer attached to peptidoglycan via a galactofuranosyl-containing polysaccharide, arabinogalactan. This structural arrangement strongly suggests that galactofuranosyl residues are essential for the growth and viability of mycobacteria. Galactofuranosyl residues are formed in nature by a ring contraction of UDP-galactopyranose to UDP-galactofuranose catalyzed by the enzyme UDP-galactopyranose mutase (Glf). In Mycobacterium tuberculosis the glf gene overlaps, by 1 nucleotide, a gene, Rv3808c, that has been shown to encode a galactofuranosyl transferase. We demonstrate here that glf can be knocked out in Mycobacterium smegmatis by allelic replacement only in the presence of two rescue plasmids carrying functional copies of glf and Rv3808c. The glf rescue plasmid was designed with a temperature-sensitive origin of replication and the M. smegmatis glf knockout mutant is unable to grow at the higher temperature at which the glf-containing rescue plasmid is lost. In a separate experiment, the Rv3808c rescue plasmid was designed with a temperature-sensitive origin of replication and the glf-bearing plasmid was designed with a normal original of replication; this strain was also unable to grow at the nonpermissive temperature. Thus, both glf and Rv3808c are essential for growth. These findings and the fact that galactofuranosyl residues are not found in humans supports the development of UDP-galactopyranose mutase and galactofuranosyl transferase as important targets for the development of new antituberculosis drugs. PMID- 11395464 TI - Rebinding of extracellular adherence protein Eap to Staphylococcus aureus can occur through a surface-bound neutral phosphatase. AB - Extracellular adherence protein Eap secreted from Staphylococcus aureus was previously found to enhance the adherence of S. aureus to eukaryotic cells. This enhancement effect is due to the ability of Eap to rebind to S. aureus and to bind to eukaryotic cells and several plasma and matrix proteins. In this study we defined one potential binding target for Eap on the surface of S. aureus, a surface-located neutral phosphatase. This phosphatase lacks an LPXTG region, but around 80% is retained on the cell surface. The soluble phosphatase can form a complex with Eap at a nonrandom molar ratio, and phosphatase activity is retained. The phosphatase can also bind to fibronectin. The cell surface-located portion presumably contributes to adherence of S. aureus to fibronectin. PMID- 11395465 TI - Global versus local regulatory roles for Lrp-related proteins: Haemophilus influenzae as a case study. AB - Lrp (leucine-responsive regulatory protein) plays a global regulatory role in Escherichia coli, affecting expression of dozens of operons. Numerous lrp-related genes have been identified in different bacteria and archaea, including asnC, an E. coli gene that was the first reported member of this family. Pairwise comparisons of amino acid sequences of the corresponding proteins shows an average sequence identity of only 29% for the vast majority of comparisons. By contrast, Lrp-related proteins from enteric bacteria show more than 97% amino acid identity. Is the global regulatory role associated with E. coli Lrp limited to enteric bacteria? To probe this question we investigated LrfB, an Lrp-related protein from Haemophilus influenzae that shares 75% sequence identity with E. coli Lrp (highest sequence identity among 42 sequences compared). A strain of H. influenzae having an lrfB null allele grew at the wild-type growth rate but with a filamentous morphology. A comparison of two-dimensional (2D) electrophoretic patterns of proteins from parent and mutant strains showed only two differences (comparable studies with lrp(+) and lrp E. coli strains by others showed 20 differences). The abundance of LrfB in H. influenzae, estimated by Western blotting experiments, was about 130 dimers per cell (compared to 3,000 dimers per E. coli cell). LrfB expressed in E. coli replaced Lrp as a repressor of the lrp gene but acted only to a limited extent as an activator of the ilvIH operon. Thus, although LrfB resembles Lrp sufficiently to perform some of its functions, its low abundance is consonant with a more local role in regulating but a few genes, a view consistent with the results of the 2D electrophoretic analysis. We speculate that an Lrp having a global regulatory role evolved to help enteric bacteria adapt to their ecological niches and that it is unlikely that Lrp related proteins in other organisms have a broad regulatory function. PMID- 11395467 TI - Biological and biochemical characterization of variant A subunits of cholera toxin constructed by site-directed mutagenesis. AB - Cholera toxin (CT) is the prototype for the Vibrio cholerae-Escherichia coli family of heat-labile enterotoxins having an AB5 structure. By substituting amino acids in the enzymatic A subunit that are highly conserved in all members of this family, we constructed 23 variants of CT that exhibited decreased or undetectable toxicity and we characterized their biological and biochemical properties. Many variants exhibited previously undescribed temperature-sensitive assembly of holotoxin and/or increased sensitivity to proteolysis, which in all cases correlated with exposure of epitopes of CT-A that are normally hidden in native CT holotoxin. Substitutions within and deletion of the entire active-site occluding loop demonstrated a prominent role for His-44 and this loop in the structure and activity of CT. Several novel variants with wild-type assembly and stability showed significantly decreased toxicity and enzymatic activity (e.g., variants at positions R11, I16, R25, E29, and S68+V72). In most variants the reduction in toxicity was proportional to the decrease in enzymatic activity. For substitutions or insertions at E29 and Y30 the decrease in toxicity was 10- and 5 fold more than the reduction in enzymatic activity, but for variants with R25G, E110D, or E112D substitutions the decrease in enzymatic activity was 12- to 50 fold more than the reduction in toxicity. These variants may be useful as tools for additional studies on the cell biology of toxin action and/or as attenuated toxins for adjuvant or vaccine use. PMID- 11395466 TI - Signal transduction cascade for regulation of RpoS: temperature regulation of DsrA. AB - Many environmental parameters modulate the amount of the RpoS sigma factor in Escherichia coli. Temperature control of RpoS depends on the untranslated RNA DsrA. DsrA activates RpoS translation by pairing with the leader of the mRNA. We find that temperature affects both the rate of transcription initiation of the dsrA gene and the stability of DsrA RNA. Both are increased at low temperature (25 degrees C) compared to 37 or 42 degrees C. The combination of these results is 25-fold-less DsrA at 37 degrees C and 30-fold less at 42 degrees C than at 25 degrees C. Using an adapted lacZ-based reporter system, we show that temperature control of transcription initiation of dsrA requires only the minimal promoter of 36 bp. Overall, transcription responses to temperature lead to a sixfold increase in DsrA synthesis at 25 degrees C over that at 42 degrees C. Furthermore, two activating regions and a site for LeuO negative regulation were identified in the dsrA promoter. The activating regions also activate transcription in vitro. DsrA decays with a half-life of 23 min at 25 degrees C and 4 min at 37 and 42 degrees C. These results demonstrate that the dsrA promoter and the stability of DsrA RNA are the thermometers for RpoS temperature sensing. Multiple inputs to DsrA accumulation allow sensitive modulation of changes in the synthesis of the downstream targets of DsrA such as RpoS. PMID- 11395468 TI - Mapping of Mycobacterium tuberculosis katG promoters and their differential expression in infected macrophages. AB - Intracellular pathogenic bacteria, including Mycobacterium tuberculosis, frequently have multitiered defense mechanisms ensuring their survival in host phagocytic cells. One such defense determinant in M. tuberculosis is the katG gene, which encodes an enzyme with catalase, peroxidase, and peroxynitritase activities. KatG is considered to be important for protection against reactive oxygen and nitrogen intermediates produced by phagocytic cells. However, KatG also activates the front-line antituberculosis drug isoniazid, hence rendering M. tuberculosis exquisitely sensitive to this compound. In this context, katG expression represents a double-edged sword, as it is an important virulence determinant but at the same time its activity levels determine sensitivity to INH. Thus, it is important to delineate the regulation and expression of katG, as this not only can aid understanding of how M. tuberculosis survives and persists in the host but also may provide information of relevance for better management of INH therapy. Here, we report the first extensive analysis of the katG promoter activity examined both in vitro and in vivo. Using S1 nuclease protection analysis, we mapped the katG mRNA 5' ends and demonstrated that two promoters, P(1)furA and P(1)katG, control transcription of katG. The furA and katG genes are cotranscribed from P(1)furA. Both P(1)furA and P(1)katG promoters show induction upon challenge with hydrogen peroxide and cumene hydroperoxide. Studies carried out using the transcriptional fusions P(1)furA-gfp, P(1)katG-gfp, and P(1)furA P(1)katG-gfp confirmed the existence of two katG promoters. In addition, we showed that both promoters are expressed in vivo during intracellular growth of virulent M. tuberculosis H37Rv. P(1)furA is induced early upon infection, and P(1)katG becomes active only upon extended growth in macrophages. These studies delineate the transcriptional organization of the furA-katG region and indicate differential regulation in vivo of the two katG promoters. These phenomena most likely reflect the differing demands at sequential stages of the infection cycle and may provide information for improved understanding of host-pathogen interactions in tuberculosis and for further optimization of INH chemotherapy. PMID- 11395469 TI - Characterization of the Ustilago maydis sid2 gene, encoding a multidomain peptide synthetase in the ferrichrome biosynthetic gene cluster. AB - Ustilago maydis, the causal agent of corn smut disease, acquires and transports ferric ion by producing the extracellular, cyclic peptide, hydroxamate siderophores ferrichrome and ferrichrome A. Ferrichrome biosynthesis likely proceeds by hydroxylation and acetylation of L-ornithine, and later steps likely involve covalently bound thioester intermediates on a multimodular, nonribosomal peptide synthetase. sid1 encodes L-ornithine N(5)-oxygenase, which catalyzes hydroxylation of L-ornithine, the first committed step of ferrichrome and ferrichrome A biosynthesis in U. maydis. In this report we characterize sid2, another biosynthetic gene in the pathway, by gene complementation, gene replacement, DNA sequence, and Northern hybridization analysis. Nucleotide sequencing has revealed that sid2 is located 3.7 kb upstream of sid1 and encodes an intronless polypeptide of 3,947 amino acids with three iterated modules of an approximate length of 1,000 amino acids each. Multiple motifs characteristic of the nonribosomal peptide synthetase protein family were identified in each module. A corresponding iron-regulated sid2 transcript of 11 kb was detected by Northern hybridization analysis. By contrast, constitutive accumulation of this large transcript was observed in a mutant carrying a disruption of urbs1, a zinc finger, GATA family transcription factor previously shown to regulate siderophore biosynthesis in Ustilago. Multiple GATA motifs are present in the intergenic region between sid1 and sid2, suggesting bidirectional transcription regulation by urbs1 of this pathway. Indeed, mutation of two of these motifs, known to be important to regulation of sid1, altered the differential regulation of sid2 by iron. PMID- 11395470 TI - Coupling of asymmetric division to polar placement of replication origin regions in Bacillus subtilis. AB - Entry into sporulation in Bacillus subtilis is characterized by the formation of a polar septum, which asymmetrically divides the developing cell into forespore (the smaller cell) and mother cell compartments, and by migration of replication origin regions to extreme opposite poles of the cell. Here we show that polar septation is closely correlated with movement of replication origins to the extreme poles of the cell. Replication origin regions were visualized by the use of a cassette of tandem copies of lacO that had been inserted in the chromosome near the origin of replication and decorated with green fluorescent protein-LacI. The results showed that extreme polar placement of replication origin regions is not under sporulation control and occurred in stationary phase under conditions under which entry into sporulation was prevented. On the other hand, the formation of a polar septum, which is under sporulation control, was almost invariably associated with the presence of a replication origin region in the forespore. Moreover, cells in which the polar placement of origin regions was perturbed by deletion of the gene (smc) for the structural maintenance of chromosomes (SMC) protein were impaired in polar division. A small proportion ( approximately 1%) of the mutant cells were able to undergo asymmetric division, but the forespore compartment of these exceptional cells was generally observed to contain a replication origin region. Immunofluorescence microscopy experiments indicated that the block in polar division caused by the absence of SMC occurred at or prior to the step of bipolar Z-ring formation by the cell division protein FtsZ. A model is discussed in which polar division is under the dual control of sporulation and an event associated with the placement of a replication origin at the cell pole. PMID- 11395471 TI - Microbial origin of plant-type 2-keto-3-deoxy-D-arabino-heptulosonate 7-phosphate synthases, exemplified by the chorismate- and tryptophan-regulated enzyme from Xanthomonas campestris. AB - Enzymes performing the initial reaction of aromatic amino acid biosynthesis, 2 keto-3-deoxy-D-arabino-heptulosonate 7-phosphate (DAHP) synthases, exist as two distinct homology classes. The three classic Escherichia coli paralogs are AroA(I) proteins, but many members of the Bacteria possess the AroA(II) class of enzyme, sometimes in combination with AroA(I) proteins. AroA(II) DAHP synthases until now have been shown to be specifically dedicated to secondary metabolism (e.g., formation of ansamycin antibiotics or phenazine pigment). In contrast, here we show that the Xanthomonas campestris AroA(II) protein functions as the sole DAHP synthase supporting aromatic amino acid biosynthesis. X. campestris AroA(II) was cloned in E. coli by functional complementation, and genes corresponding to two possible translation starts were expressed. We developed a 1 day partial purification method (>99%) for the unstable protein. The recombinant AroA(II) protein was found to be subject to an allosteric pattern of sequential feedback inhibition in which chorismate is the prime allosteric effector. L Tryptophan was found to be a minor feedback inhibitor. An N-terminal region of 111 amino acids may be located in the periplasm since a probable inner membrane spanning region is predicted. Unlike chloroplast-localized AroA(II) of higher plants, X. campestris AroA(II) was not hysteretically activated by dithiols. Compared to plant AroA(II) proteins, differences in divalent metal activation were also observed. Phylogenetic tree analysis shows that AroA(II) originated within the Bacteria domain, and it seems probable that higher-plant plastids acquired AroA(II) from a gram-negative bacterium via endosymbiosis. The X. campestris AroA(II) protein is suggested to exemplify a case of analog displacement whereby an ancestral aroA(I) species was discarded, with the aroA(II) replacement providing an alternative pattern of allosteric control. Three subgroups of AroA(II) proteins can be recognized: a large, central group containing the plant enzymes and that from X. campestris, one defined by a three residue deletion near the conserved KPRS motif, and one possessing a larger deletion further downstream. PMID- 11395472 TI - In vivo evidence for two active nuclease motifs in the double-strand break repair enzyme RexAB of Lactococcus lactis. AB - In bacteria, double-strand DNA break (DSB) repair involves an exonuclease/helicase (exo/hel) and a short regulatory DNA sequence (Chi) that attenuates exonuclease activity and stimulates DNA repair. Despite their key role in cell survival, these DSB repair components show surprisingly little conservation. The best-studied exo/hel, RecBCD of Escherichia coli, is composed of three subunits. In contrast, RexAB of Lactococcus lactis and exo/hel enzymes of other low-guanine-plus-cytosine branch gram-positive bacteria contain two subunits. We report that RexAB functions via a novel mechanism compared to that of the RecBCD model. Two potential nuclease motifs are present in RexAB compared with a single nuclease in RecBCD. Site-specific mutagenesis of the RexA nuclease motif abolished all nuclease activity. In contrast, the RexB nuclease motif mutants displayed strongly reduced nuclease activity but maintained Chi recognition and had a Chi-stimulated hyperrecombination phenotype. The distinct phenotypes resulting from RexA or RexB nuclease inactivation lead us to suggest that each of the identified active nuclease sites in RexAB is involved in the degradation of one DNA strand. In RecBCD, the single RecB nuclease degrades both DNA strands and is presumably positioned by RecD. The presence of two nucleases would suggest that this RecD function is dispensable in RexAB. PMID- 11395474 TI - Efg1, a morphogenetic regulator in Candida albicans, is a sequence-specific DNA binding protein. AB - Efg1 is essential for hyphal development in the human pathogen Candida albicans under most conditions. Efg1 is related to basic helix-loop-helix regulators, and therefore most workers presume that Efg1 is a transcription factor. Here we confirm that Efg1 is a DNA binding protein that can interact specifically with the E box. PMID- 11395473 TI - Efficient vir gene induction in Agrobacterium tumefaciens requires virA, virG, and vir box from the same Ti plasmid. AB - The vir genes of octopine, nopaline, and L,L-succinamopine Ti plasmids exhibit structural and functional similarities. However, we observed differences in the interactions between octopine and nopaline vir components. The induction of an octopine virE(A6)::lacZ fusion (pSM358cd) was 2.3-fold higher in an octopine strain (A348) than in a nopaline strain (C58). Supplementation of the octopine virG(A6) in a nopaline strain with pSM358 did not completely restore virE(A6) induction. However, addition of the octopine virA(A6) to the above strain increased virE(A6) induction to a level almost comparable to that in octopine strains. In a reciprocal analysis, the induction of a nopaline virE(C58)::cat fusion (pUCD1553) was two- to threefold higher in nopaline (C58 and T37) strains than in octopine (A348 and Ach5) and L,L-succinamopine (A281) strains. Supplementation of nopaline virA(C58) and virG(C58) in an octopine strain (A348) harboring pUCD1553 increased induction levels of virE(C58)::cat fusion to a level comparable to that in a nopaline strain (C58). Our results suggest that octopine and L,L-succinamopine VirG proteins induce the octopine virE(A6) more efficiently than they do the nopaline virE(C58). Conversely, the nopaline VirG protein induces the nopaline virE(C58) more efficiently than it does the octopine virE(A6). The ability of Bo542 virG to bring about supervirulence in tobacco is observed for an octopine vir helper (LBA4404) but not for a nopaline vir helper (PMP90). Our analyses reveal that quantitative differences exist in the interactions between VirG and vir boxes of different Ti plasmids. Efficient vir gene induction in octopine and nopaline strains requires virA, virG, and vir boxes from the respective Ti plasmids. PMID- 11395475 TI - DNA-binding activity of amino-terminal domains of the Bacillus subtilis AbrB protein. AB - Two truncated variants of AbrB, comprising either its first 53 (AbrBN53) or first 55 (AbrBN55) amino acid residues, were constructed and purified. Noncovalently linked homodimers of the truncated variants exhibited very weak DNA-binding activity. Cross-linking AbrBN55 dimers into tetramers and higher-order multimers (via disulfide bonding between penultimate cysteine residues) resulted in proteins having DNA-binding affinity comparable to and DNA-binding specificity identical to those of intact, wild-type AbrB. These results indicate that the DNA recognition and specificity determinants of AbrB binding lie solely within its N terminal amino acid sequence. PMID- 11395476 TI - Characterization of Escherichia coli type 1 pilus mutants with altered binding specificities. AB - PCR mutagenesis and a unique enrichment scheme were used to obtain two mutants, each with a single lesion in fimH, the chromosomal gene that encodes the adhesin protein (FimH) of Escherichia coli type 1 pili. These mutants were noteworthy in part because both were altered in the normal range of cell types bound by FimH. One mutation altered an amino acid at a site previously shown to be involved in temperature-dependent binding, and the other altered an amino acid lining the predicted FimH binding pocket. PMID- 11395477 TI - The catalytic function of bovine lysyl oxidase in the absence of copper. AB - Bovine lysyl oxidase (BLO) contains two different cofactors, copper (Kagan, H. M. (1986) in Biology of Extracellular Matrix (Mecham, R. P., ed) Vol. 1, pp. 321 398, Academic Press, Orlando, FL) and lysine tyrosyl quinone (LTQ) (Wang, S. X., Mure, M., Medzihradszky, K. F., Burlingame, A. L., Brown, D. E., Dooley, D. M., Smith, A. J., Kagan, H. M., and Klinman, J. P. (1996) Science 273, 1078-1084). By a combination of UV-visible spectroscopy, metal content analysis, and activity measurements, we find that copper-depleted BLO reacts in an irreversible manner with phenylhydrazine, an amine substrate analog, and catalyzes multiple turnovers of the substrate benzylamine. After removal of the majority of enzyme-bound copper, apoBLO exhibits a decrease in the LTQ content, as evidenced by the drop of the 510-520-nm absorbance, suggesting that the copper may play a structural role in stabilizing the LTQ. The remaining intact LTQ in the apoBLO reacted with phenylhydrazine, both in the presence and absence of the chelator, 10 mm 2,2' dipyridyl. When benzylamine was used as the substrate, the apoBLO turned over at a rate of 50-60% of the native BLO (after correction for the residual copper and the change of LTQ content). Copper contamination from the assay buffer was ruled out by comparison of enzyme activity using different apoBLO concentrations. These studies demonstrate that the mature form of lysyl oxidase retains many of its functions in the absence of copper. PMID- 11395478 TI - Nuclear localization and dominant-negative suppression by a mutant SKCa3 N terminal channel fragment identified in a patient with schizophrenia. AB - The small conductance calcium-activated K+ channel gene SKCa3/KCNN3 maps to 1q21, a region strongly linked to schizophrenia. Recently, a 4-base pair deletion in SKCa3 was reported in a patient with schizophrenia, which truncates the protein at the end of the N-terminal cytoplasmic region (SKCa3Delta). We generated a green fluorescent protein-SKCa3 N-terminal construct (SKCa3-1/285) that is identical to SKCa3Delta except for the last two residues. Using confocal microscopy we demonstrate that SKCa3-1/285 localizes rapidly and exclusively to the nucleus of mammalian cells like several other pathogenic polyglutamine containing proteins. This nuclear targeting is mediated in part by two polybasic sequences present at the C-terminal end of SKCa3-1/285. In contrast, full-length SKCa3, SKCa2, and IKCa1 polypeptides are all excluded from the nucleus and express as functional channels. When overexpressed in human Jurkat T cells, SKCa3 1/285 can suppress endogenous SKCa2 currents but not voltage-gated K+ currents. This dominant-negative suppression is most likely mediated through the co assembly of SKCa3-1/285 with native subunits and the formation of non-functional tetramers. The nuclear localization of SKCa3-1/285 may alter neuronal architecture, and its ability to dominantly suppress endogenous small conductance K(Ca) currents may affect patterns of neuronal firing. Together, these two effects may play a part in the pathogenesis of schizophrenia and other neuropsychiatric disorders. PMID- 11395479 TI - SET-related cell division autoantigen-1 (CDA1) arrests cell growth. AB - We used an autoimmune serum from a patient with discoid lupus erythematosus to clone a cDNA of 2808 base pairs. Its open reading frame of 2079 base pairs encodes a predicted polypeptide of 693 amino acids named CDA1 (cell division autoantigen-1). CDA1 has a predicted molecular mass of 79,430 Daltons and a pI of 4.26. The size of the cDNA is consistent with its estimated mRNA size. CDA1 comprises an N-terminal proline-rich domain, a central basic domain, and a C terminal bipartite acidic domain. It has four putative nuclear localization signals and potential sites for phosphorylation by cAMP and cGMP-dependent kinases, protein kinase C, thymidine kinase, casein kinase II, and cyclin dependent kinases (CDKs). CDA1 is phosphorylated in HeLa cells and by cyclin D1/CDK4, cyclin A/CDK2, and cyclin B/CDK1 in vitro. Its basic and acidic domains contain regions homologous to almost the entire human leukemia-associated SET protein. The same basic region is also homologous to nucleosome assembly proteins, testis TSPY protein, and an uncharacterized brain protein. CDA1 is present in the nuclear fraction of HeLa cells and localizes to the nucleus and nucleolus in HeLa cells transfected with CDA1 or its N terminus containing all four nuclear localization signals. Its acidic C terminus localizes mainly to the cytoplasm. CDA1 levels are low in serum-starved cells, increasing dramatically with serum stimulation. Expression of the CDA1 transgene, but not its N terminus, arrests HeLa cell growth, colony numbers, cell density, and bromodeoxyuridine uptake in a dose-dependent manner. The ability of CDA1 to arrest cell growth is abolished by mutation of the two CDK consensus phosphorylation sites. We propose that CDA1 is a negative regulator of cell growth and that its activity is regulated by its expression level and phosphorylation. PMID- 11395482 TI - Human Ca2+/calmodulin-dependent protein kinase kinase beta gene encodes multiple isoforms that display distinct kinase activity. AB - Ca(+2)/calmodulin-dependent protein kinases (CaMKs) are activated upon binding of Ca(+2)/calmodulin. To gain maximal activity, CaMK I and CaMK IV can be further phosphorylated by an upstream kinase, CaMK kinase (CaMKK). We previously isolated cDNA clones encoding human CaMKK beta isoforms that are heterogeneous in their 3' sequences (Hsu, L.-S., Tsou, A.-P., Chi, C.-W., Lee, C.-H., and Chen, J.-Y. (1998) J. Biomed. Sci. 5, 141-149). In the present study, we examined the genomic organization and transcription of the human CaMKK beta gene. The human CaMKK beta locus spans more than 40 kilobase pairs and maps to chromosome 12q24.2. It is organized into 18 exons and 17 introns that are flanked by typical splice donor and acceptor sequences. Two major species of transcripts, namely the beta1 (5.6 kilobase pairs) and beta2 (2.9 kilobase pairs), are generated through differential usage of polyadenylation sites located in the last and penultimate exons. Additional forms of CaMKK beta transcripts were also identified that resulted from alternative splicing of the internal exons 14 and/or 16. These isoforms display differential expression patterns in human tissues and tumor derived cell lines. They also exhibit a distinct ability to undergo autophosphorylation and to phosphorylate the downstream kinases CaMK I and CaMK IV. The differential expression of CaMKK beta isoforms with distinct activity further suggests the complexity of the regulation of the CaMKK/CaMK cascade and an important role for CaMKK in the action of Ca(+2)-mediated cellular responses. PMID- 11395480 TI - Bone morphogenetic protein-2 promotes osteoblast apoptosis through a Smad independent, protein kinase C-dependent signaling pathway. AB - Bone morphogenetic protein-2 (BMP-2), a member of the transforming growth factor beta (TGF-beta) family, regulates osteoblast differentiation and bone formation. Here we show a novel function of BMP-2 in human osteoblasts and identify a signaling pathway involved in this function. BMP-2 promotes apoptosis in primary human calvaria osteoblasts and in immortalized human neonatal calvaria osteoblasts, as shown by terminal deoxynucleotidyl transferase-mediated nick end labeling analysis. In contrast, TGF-beta 2 inhibits apoptosis in human osteoblasts. Studies of the mechanisms of action showed that BMP-2 increases the Bax/Bcl-2 ratio, whereas TG beta-2 has a negative effect. Moreover, BMP-2 increases the release of mitochondrial cytochrome c to the cytosol. Consistent with these results, BMP-2 increases caspase-9 and caspase-3, -6, and -7 activity, and an anti-caspase-9 agent suppresses BMP-2-induced apoptosis. Overexpression of dominant-negative Smad1 effectively blocks BMP-2-induced expression of the osteoblast transcription factor Runx2 but not the activation of caspases or apoptosis induced by BMP-2, indicating that the Smad1 signaling pathway is not involved in the BMP-2-induced apoptosis. The proapoptotic effect of BMP-2 is PKC dependent, because BMP-2 increases PKC activity, and the selective PKC inhibitor calphostin C blocks the BMP-2-induced increased Bax/Bcl-2, caspase activity, and apoptosis. In contrast, the cAMP-dependent protein kinase A inhibitor H89, the p38 MAPK inhibitor SB203580, and the MEK inhibitor PD-98059 have no effect. The results show that BMP-2 uses a Smad-independent, PKC-dependent pathway to promote apoptosis via a Bax/Bcl-2 and cytochrome c-caspase-9-caspase-3, -6, -7 cascade in human osteoblasts. PMID- 11395481 TI - Molecular mass, stoichiometry, and assembly of 20 S particles. AB - N-Ethylmaleimide-sensitive factor (NSF), soluble NSF attachment proteins (SNAPs), and SNAP receptor (neuronal SNARE) complexes form 20 S particles with a mass of 788 +/- 122 kDa as judged by scanning transmission electron microscopy. A single NSF hexamer and three alpha SNAP monomers reside within a 20 S particle as determined by quantitative amino acid analysis. In order to study the binding of alpha SNAP and NSF in solution, to define their binding domains, and to specify the role of oligomerization in their interaction, we fused domains of alpha SNAP and NSF to oligomerization modules derived from thrombospondin-1, a trimer, and cartilage oligomeric matrix protein, a pentamer, respectively. Binding studies with these fusion proteins reproduced the interaction of alpha SNAP and NSF N domains in the absence of the hexamerization domain of NSF (D2). Trimeric alpha SNAP (or its C-terminal half) is sufficient to recruit NSF even in the absence of SNARE complexes. Furthermore, pentameric NSF N domains are able to bind alpha SNAP in complex with SNAREs, whereas monomeric N domains do not. Our results demonstrate that the oligomerization of both NSF N domains and alpha SNAP provides a critical driving force for their interaction and the assembly of 20 S particles. PMID- 11395483 TI - The uptake inhibitors cocaine and benztropine differentially alter the conformation of the human dopamine transporter. AB - The binding affinity of the cocaine analog [(3)H]2 beta-carbomethoxy-3beta-(4 fluorophenyl) tropane (WIN) for the dopamine transporter (DAT) is increased by the reaction of Cys-90, at the extracellular end of the first transmembrane segment, with methanethiosulfonate (MTS) reagents. Cocaine enhances the reaction of Cys-90 with the sulfhydryl reagents, thereby augmenting the increase in binding. In contrast, cocaine decreases the reaction of Cys-135 and Cys-342, endogenous cysteines in cytoplasmic loops, with MTS reagents. Because this reaction inhibits [(3)H]WIN binding, cocaine protects against the loss of binding caused by reaction of these cysteines. In the present work, we compare the abilities of DAT inhibitors and substrates to affect the reaction of Cys-90, Cys 135, and Cys-342 with MTS ethyltrimethylammonium (MTSET). The results indicate that the different abilities of compounds to protect against the MTSET-induced inhibition of binding are attributable to differences in their abilities to attenuate the inhibitory effects of modification of Cys-135 and Cys-342 as well as to enhance the reaction with Cys-90 and the resulting potentiation of binding. The inhibitor benztropine was unique in its inability to protect Cys-135. Moreover, whereas cocaine, WIN, mazindol, and dopamine enhanced the reaction of Cys-90 with MTSET, benztropine had no effect on this reaction. These two features combine to give benztropine its weak potency in protecting ligand binding to wild type DAT from MTSET. These results indicate that different inhibitors of DAT, such as cocaine and benztropine, produce different conformational changes in the transporter. There are differences in the psychomotor stimulant-like effects of these compounds, and it is possible that the different behavioral effects of these DAT inhibitors stem from their different molecular actions on DAT. PMID- 11395484 TI - Opposing effects of molecular volume and charge at the hyperekplexia site alpha 1(P250) govern glycine receptor activation and desensitization. AB - Allelic variants of the glycine receptor alpha1 subunit gene GLRA1 underlie the human neurological disorder hyperekplexia. Among these, the subunit variant alpha1(P250T) is characterized by an amino acid substitution within the cytoplasmic TM1-2 loop. To identify structural elements at position alpha1(250) that govern receptor function, homomeric mutant receptor channels were subjected to electrophysiological analysis after recombinant expression in HEK293 cells. Wild-type alpha1(P250) channels were nondesensitizing with an EC(50) for glycine of 8 microm, whereas bulky hydrophobic side chains of the channel variants alpha1(P250V/I/L/F) showed rapid desensitization (tau(desens), 50-250 ms) and EC(50) values of 400-1800 microm. Small side chains (P250G/A/S) gave rise to wild type-like channels. Effects of volume were counteracted by charge: alpha1(P250E/R) were nondesensitizing; EC(50) was approximately 70 microm. The mutants alpha1(P250C/Y) displayed intermediate channel properties (EC(50), 42/70 microm; tau(desens), 3300/2800 ms, respectively). The isotropic forces volume and hydropathy were sufficient to account for the observed effects of residue alpha1(250) on receptor function. Indeed, channel behavior was best predicted by a combined hydropathy/volume index describing the hydrophobic surface of individual amino acids. These observations characterize the short intracellular TM1-2 loop as a regulatory domain for channel activation and a crucial mediator of glycine receptor desensitization. PMID- 11395485 TI - Coupled electron/proton transfer in complex flavoproteins: solvent kinetic isotope effect studies of electron transfer in xanthine oxidase and trimethylamine dehydrogenase. AB - A solvent kinetic isotope effect study of electron transfer in two complex flavoproteins, xanthine oxidase and trimethylamine dehydrogenase, has been undertaken. With xanthine oxidase, electron transfer from the molybdenum center to the proximal iron-sulfur center of the enzyme occurs with a modest solvent kinetic isotope effect of 2.2, indicating that electron transfer out of the molybdenum center is at least partially coupled to deprotonation of the Mo(V) donor. A Marcus-type analysis yields a decay factor, beta, of 1.4 A(-1), indicating that, although the pyranopterin cofactor of the molybdenum center forms a nearly contiguous covalent bridge from the molybdenum atom to the proximal iron-sulfur center of the enzyme, it affords no exceptionally effective mode of electron transfer between the two centers. For trimethylamine dehydrogenase, rates of electron equilibration between the flavin and iron-sulfur center of the one-electron reduced enzyme have been determined, complementing previous studies of electron transfer in the two-electron reduced form. The results indicate a substantial solvent kinetic isotope effect of 10 +/- 4, consistent with a model for electron transfer that involves discrete protonation/deprotonation and electron transfer steps. This contrasts to the behavior seen with xanthine oxidase, and the basis for this difference is discussed in the context of the structures for the two proteins and the ionization properties of their flavin sites. With xanthine oxidase, a rationale is presented as to why it is desirable in certain cases that the physical layout of redox-active sites not be uniformly increasing in reduction potential in the direction of physiological electron transfer. PMID- 11395486 TI - The LG3 module of laminin-5 harbors a binding site for integrin alpha3beta1 that promotes cell adhesion, spreading, and migration. AB - Laminins are a family of extracellular matrix glycoproteins involved in cell adhesion and migration. A major obstacle to understanding their structure function relationships is the lack of small laminin domains capable of replicating integrin-binding, cell-adhesive, and migratory functions of the intact molecule. Here, we show that the recombinant LG3 (rLG3) module (26 kDa) of laminin-5 (Ln-5) alpha(3) chain replicated key Ln-5 activities. rLG3 but not rLG1 or rLG2 supported cell adhesion and migration of at least two distinct cell lines, in an integrin alpha(3)beta(1)-dependent manner. Cell adhesion to rLG3 was regulated by divalent cations and accompanied by cell spreading and tyrosine phosphorylation of FAK focal adhesion kinase. The integrin binding activity of rLG3 was confirmed by rLG3 affinity chromatography of detergent cell lysates, which resulted in specific purification of integrin alpha(3)beta(1). To our knowledge, this is the first report directly demonstrating that a recombinant laminin LG module is an active domain capable of supporting integrin-dependent cell adhesion and migration. PMID- 11395487 TI - The tip of the coiled-coil rod determines the filament formation of smooth muscle and nonmuscle myosin. AB - Myosin II self-assembles to form thick filaments that are attributed to its long coiled-coil tail domain. The present study has determined a region critical for filament formation of vertebrate smooth muscle and nonmuscle myosin II. A monoclonal antibody recognizing the 28 residues from the C-terminal end of the coiled-coil domain of smooth muscle myosin II completely inhibited filament formation, whereas other antibodies recognizing other parts of the coiled-coil did not. To determine the importance of this region in the filament assembly in vivo, green fluorescent protein (GFP)-tagged smooth muscle myosin was expressed in COS-7 cells, and the filamentous localization of the GFP signal was monitored by fluorescence microscopy. Wild type GFP-tagged smooth muscle myosin colocalized with F-actin during interphase and was also recruited into the contractile ring during cytokinesis. Myosin with the nonhelical tail piece deleted showed similar behavior, whereas deletion of the 28 residues at the C-terminal end of the coiled coil domain abolished this localization. Deletion of the corresponding region of GFP-tagged nonmuscle myosin IIA also abolished this localization. We conclude that the C-terminal end of the coiled-coil domain, but not the nonhelical tail piece, of myosin II is critical for myosin filament formation both in vitro and in vivo. PMID- 11395488 TI - Sucrase-isomaltase gene transcription requires the hepatocyte nuclear factor-1 (HNF-1) regulatory element and is regulated by the ratio of HNF-1 alpha to HNF-1 beta. AB - The mouse sucrase-isomaltase (SI) gene is an enterocyte-specific gene expressed in a complex developmental pattern. We previously reported that a short, evolutionarily conserved gene promoter regulates developmental expression of SI in mouse small intestine. Herein, we investigated the role of a hepatocyte nuclear factor-1 (HNF-1) cis-acting element to regulate SI gene expression in vivo. Transgenic SI gene constructs with a mutated HNF-1 element (SIF3) revealed a strong reduction in promoter activity in comparison with a wild-type construct in mice and during Caco-2 cell differentiation. Nuclear proteins isolated from enterocytes showed increased binding of the HNF-1 alpha complex with a concomitant decrease in the HNF-1 beta-containing complex to the SIF3 element both during the suckling-weaning developmental transition and Caco-2 cell differentiation. These changes coincided with a strong induction of SI gene transcription. In transfection experiments, HNF-1 alpha activated the SI promoter via the SIF3 element, and co-expression of HNF-1 beta impaired this transcriptional activation. These findings demonstrate the essential role of the HNF-1 regulatory element to support SI gene transcription in vivo and suggest that the ratio of HNF-1 alpha to HNF-1 beta plays a role in the transcriptional activity of this gene during intestinal development. PMID- 11395489 TI - Chicken avidin exhibits pseudo-catalytic properties. Biochemical, structural, and electrostatic consequences. AB - Avidin and its bacterial analogue streptavidin exhibit similarly high affinities toward the vitamin biotin. The extremely high affinity of these two proteins has been utilized as a powerful tool in many biotechnological applications. Although avidin and streptavidin have similar tertiary and quaternary structures, they differ in many of their properties. Here we show that avidin enhances the alkaline hydrolysis of biotinyl p-nitrophenyl ester, whereas streptavidin protects this reaction even under extreme alkaline conditions (pH > 12). Unlike normal enzymatic catalysis, the hydrolysis reaction proceeds as a single cycle with no turnover because of the extremely high affinity of the protein for one of the reaction products (i.e. free biotin). The three-dimensional crystal structures of avidin (2 A) and streptavidin (2.4 A) complexed with the amide analogue, biotinyl p-nitroanilide, as a model for the p-nitrophenyl ester, revealed structural insights into the factors that enhance or protect the hydrolysis reaction. The data demonstrate that several molecular features of avidin are responsible for the enhanced hydrolysis of biotinyl p-nitrophenyl ester. These include the nature of a decisive flexible loop, the presence of an obtrusive arginine 114, and a newly formed critical interaction between lysine 111 and the nitro group of the substrate. The open conformation of the loop serves to expose the substrate to the solvent, and the arginine shifts the p nitroanilide moiety toward the interacting lysine, which increases the electron withdrawing characteristics and consequent electrophilicity of the carbonyl group of the substrate. Streptavidin lacked such molecular properties, and analogous interactions with the substrate were consequently absent. The information derived from these structures may provide insight into the action of artificial protein catalysts and the evolution of catalytic sites in general. PMID- 11395490 TI - Characterization of a novel complex from halophilic archaebacteria, which displays chaperone-like activities in vitro. AB - We isolated a protein, P45, from the extreme halophilic archaeon Haloarcula marismortui, which displays molecular chaperone activities in vitro. P45 is a weak ATPase that assembles into a large ring-shaped oligomeric complex comprising about 10 subunits. The protein shows no significant homology to any known protein. P45 forms complexes with halophilic malate dehydrogenase during its salt dependent denaturation/renaturation and decreases the rate of deactivation of the enzyme in an ATP-dependent manner. Compared with other halophilic proteins, the P45 complex appears to be much less dependent on salt for its various activities or stability. In vivo experiments showed that P45 accumulates when cells are exposed to a low salt environment. We suggest, therefore, that P45 could protect halophilic proteins against denaturation under conditions of cellular hyposaline stress. PMID- 11395491 TI - Identification of the minimal tyrosine residues required for linker for activation of T cell function. AB - The linker for activation of T cells (LAT) is essential for signaling through the T cell receptor (TCR). Following TCR stimulation, LAT becomes tyrosine phosphorylated, creating docking sites for other signaling proteins such as phospholipase C-gamma(1) (PLC-gamma(1)), Grb2, and Gads. In this study, we have attempted to identify the critical tyrosine residues in LAT that mediate TCR activation-induced mobilization of intracellular Ca(2+) and activation of the MAP kinase Erk2. By using the LAT-deficient Jurkat derivative, J.CaM2, stable cell lines were established expressing various tyrosine mutants of LAT. We show that three specific tyrosine residues (Tyr(132), Tyr(171), and Tyr(191)) are necessary and sufficient to achieve a Ca(2+) flux following TCR stimulation. These tyrosine residues function by reconstituting PLC-gamma(1) phosphorylation and recruitment to LAT. However, these same tyrosines can only partially reconstitute Erk activation. Full reconstitution of Erk requires two additional tyrosine residues (Tyr(110) and Tyr(226)), both of which have the Grb2-binding motif YXN. This reconstitution of Erk activation requires that the critical tyrosine residues be on the same molecule of LAT, suggesting that a single LAT molecule nucleates multiple protein-protein interactions required for optimal signal transduction. PMID- 11395492 TI - Photosynthetic electron transfer through the cytochrome b6f complex can bypass cytochrome f. AB - The cytochrome b(6)f complex is an obligatory electron transfer and proton translocating enzyme in all oxygenic photosynthesis. Its operation has been described by the "Q-cycle." This model proposes that electrons are transferred from plastoquinol to plastocyanin (the reductant of P700 in Photosystem I) through, obligatorily in series, the iron-sulfur and the cytochrome f redox centers in the cytochrome b(6)f complex. However, here we demonstrate that (a) the iron-sulfur center-dependent reductions of plastocyanin and P700 are much faster than cytochrome f reduction, both in Chlamydomonas reinhardtii cytochrome f mutants and in the wild type, and (b) the steady-state photosynthetic electron transport does not correlate with strongly inhibited cytochrome f reduction kinetics in the mutants. Thus, cytochrome f is not an obligatory intermediate for electrons flowing through the cytochrome b(6)f complex. The oxidation equivalents from Photosystem I are delivered to the high potential chain of the cytochrome b(6)f complex both at the cytochrome f level and, independently, at another site connected to the quinol-oxidizing site, possibly the iron-sulfur center. PMID- 11395493 TI - BRCT domain-containing protein TopBP1 functions in DNA replication and damage response. AB - Topoisomerase IIbeta-binding protein (TopBP1), a human protein with eight BRCT domains, is similar to Saccharomyces cerevisiae Dpb11 and Schizosaccharomyces pombe Cut5 checkpoint proteins and closely related to Drosophila Mus101. We show that human TopBP1 is required for DNA replication and that it interacts with DNA polymerase epsilon. In S phase TopBP1 colocalizes with Brca1 to foci that do not represent sites of ongoing DNA replication. Inhibition of DNA synthesis leads to relocalization of TopBP1 together with Brca1 to replication forks, suggesting a role in rescue of stalled forks. DNA damage induces formation of distinct TopBP1 foci that colocalize with Brca1 in S phase, but not in G(1) phase. We also show that TopBP1 interacts with the checkpoint protein hRad9. Thus, these results implicate TopBP1 in replication and checkpoint functions. PMID- 11395494 TI - Identification of Sam68 arginine glycine-rich sequences capable of conferring nonspecific RNA binding to the GSG domain. AB - Sam68 is an RNA-binding protein that contains a heterogeneous nuclear ribonucleoprotein K homology domain embedded in a larger RNA binding domain called the GSG (GRP33, Sam68, GLD-1) domain. This family of proteins is often referred to as the STAR (signal transduction and activators of RNA metabolism) proteins. It is not known whether Sam68 is a general nonspecific RNA-binding protein or whether it recognizes specific response elements in mRNAs with high affinity. Sam68 has been shown to bind homopolymeric RNA and a synthetic RNA sequence called G8-5 that has a core UAAA motif. Here we performed a structure function analysis of Sam68 and identified two arginine glycine (RG)-rich regions that confer nonspecific RNA binding to the Sam68 GSG domain. In addition, by using chimeric proteins between Sam68 and QKI-7, we demonstrated that one of the Sam68 RG-rich sequences of 26 amino acids was sufficient to confer homopolymeric RNA binding to the GSG domain of QKI-7, another STAR protein. Furthermore, that minimal sequence can also give QKI-7 the ability (as Sam68) to functionally substitute for HIV-1 REV to facilitate the nuclear export of RNAs. Our studies suggest that neighboring RG-rich sequences may impose nonspecific RNA binding to GSG domains. Because the Sam68 RNA binding activity is negatively regulated by tyrosine phosphorylation, our data lead us to propose that Sam68 might be a specific RNA-binding protein when tyrosine phosphorylated. PMID- 11395495 TI - Dynamic sensitivity of ATP-sensitive K(+) channels to ATP. AB - ATP and MgADP regulate K(ATP) channel activity and hence potentially couple cellular metabolism to membrane electrical activity in various cell types. Using recombinant K(ATP) channels that lack sensitivity to MgADP, expressed in COSm6 cells, we demonstrate that similar on-cell activity can be observed with widely varying apparent submembrane [ATP] ([ATP](sub)). Metabolic inhibition leads to a biphasic change in the channel activity; activity first increases, presumably in response to a fast decrease in [ATP](sub), and then declines. The secondary decrease in channel activity reflects a marked increase in ATP sensitivity and is correlated with a fall in polyphosphoinositides (PPIs), including phosphatidylinositol 4,5-bisphosphate, probed using equilibrium labeling of cells with [(3)H]myo-inositol. Both ATP sensitivity and PPIs rapidly recover following removal of metabolic inhibition, and in both cases recovery is blocked by wortmannin. These data are consistent with metabolism having a dual effect on K(ATP) channel activity: rapid activation of channels because of relief of ATP inhibition and much slower reduction of channel activity mediated by a fall in PPIs. These two mechanisms constitute a feedback system that will tend to render K(ATP) channel activity transiently responsive to a change in [ATP](sub) over a wide range of steady state concentrations. PMID- 11395497 TI - Mapping of calmodulin and Gbetagamma binding domains within the C-terminal region of the metabotropic glutamate receptor 7A. AB - Ca(2+)/calmodulin (Ca(2+)/CaM) and the betagamma subunits of heterotrimeric G proteins (Gbetagamma) have recently been shown to interact in a mutually exclusive fashion with the intracellular C terminus of the presynaptic metabotropic glutamate receptor 7 (mGluR 7). Here, we further characterized the core CaM and Gbetagamma binding sequences. In contrast to a previous report, we find that the CaM binding motif localized in the N-terminal region of the cytoplasmic tail domain of mGluR 7 is conserved in the related group III mGluRs 4A and 8 and allows these receptors to also bind Ca(2+)/CaM. Mutational analysis of the Ca(2+)/CaM binding motif is consistent with group III receptors containing a conventional CaM binding site formed by an amphipathic alpha-helix. Substitutions adjacent to the core CaM target sequence selectively prevent Gbetagamma binding, suggesting that the CaM-dependent regulation of signal transduction involves determinants that overlap with but are different from those mediating Gbetagamma recruitment. In addition, we present evidence that Gbetagamma uses distinct nonoverlapping interfaces for interaction with the mGluR 7 C-terminal tail and the effector enzyme adenylyl cyclase II, respectively. Although Gbetagamma-mediated signaling is abolished in receptors lacking the core CaM binding sequence, alpha subunit activation, as assayed by agonist-dependent GTPgammaS binding, was not affected. This suggests that Ca(2+)/CaM may alter the mode of group III mGluR signaling from mono- (alpha) to bidirectional (alpha and betagamma) activation of downstream effector cascades. PMID- 11395496 TI - Activation of matrix metalloproteinases by peroxynitrite-induced protein S glutathiolation via disulfide S-oxide formation. AB - Oxidative stress may cause tissue injury through activation of the precursors of matrix metalloproteinase (proMMPs). In this study, we observed glutathione (GSH) dependent proMMP activation induced by peroxynitrite, a potent oxidizing agent formed during inflammatory processes. Peroxynitrite strongly activated all three types of purified human proMMPs (proMMP-1, -8, and -9) in the presence of similar concentrations of GSH. Of the potential reaction products between peroxynitrite and GSH, only S-nitroglutathione (GSNO(2)) caused proMMP activation. Extensive S glutathiolation of the proMMP protein occurred during activation of proMMP by peroxynitrite and GSH, as shown by radiolabeling studies with [(35)S]GSH or [(3)H]GSH. Evidence of appreciable S-glutathiolation persisted even after dithiothreitol and protein-denaturing treatment, however, suggesting that some S glutathiolation did not occur through formation of simple mixed disulfide. Matrix assisted laser-desorption ionization-time-of-flight mass spectrometry indicated that not only peroxynitrite plus GSH but also synthetic GSNO(2) produced dithiothreitol-resistant S-glutathiolation of the synthetic peptide PRCGVPD, which is a well conserved Cys-containing sequence of the propeptide autoinhibitory domain of proMMPs. PRCGVPD S-glutathiolation is presumed to be formed through glutathione disulfide S-oxide (GS(O)SR), based on the m/z 1064. Our results illustrate a unique mechanism of oxidative proMMP activation and oxidative tissue injury during inflammation. PMID- 11395498 TI - The disordered mobile loop of GroES folds into a defined beta-hairpin upon binding GroEL. AB - The GroES mobile loop is a stretch of approximately 16 amino acids that exhibits a high degree of flexible disorder in the free protein. This loop is responsible for the interaction between GroES and GroEL, and it undergoes a folding transition upon binding to GroEL. Results derived from a combination of transferred nuclear Overhauser effect NMR experiments and molecular dynamics simulations indicate that the mobile loop adopts a beta-hairpin structure with a Type I, G1 Bulge turn. This structure is distinct from the conformation of the loop in the co-crystal of GroES with GroEL-ADP but identical to the conformation of the bacteriophage-panned "strongly binding peptide" in the co-crystal with GroEL. Analysis of sequence conservation suggests that sequences of the mobile loop and strongly binding peptide were selected for the ability to adopt this hairpin conformation. PMID- 11395499 TI - Structural evidence for a functional role of human tissue nonspecific alkaline phosphatase in bone mineralization. AB - The human tissue nonspecific alkaline phosphatase (TNAP) is found in liver, kidney, and bone. Mutations in the TNAP gene can lead to Hypophosphatasia, a rare inborn disease that is characterized by defective bone mineralization. TNAP is 74% homologous to human placental alkaline phosphatase (PLAP) whose crystal structure has been recently determined at atomic resolution (Le Du, M. H., Stigbrand, T., Taussig, M. J., Menez, A., and Stura, E. A. (2001) J. Biol. Chem, 276, 9158-9165). The degree of homology allowed us to build a reliable TNAP model to investigate the relationship between mutations associated with hypophosphatasia and their probable consequences on the activity or the structure of the enzyme. The mutations are clustered within five crucial regions, namely the active site and its vicinity, the active site valley, the homodimer interface, the crown domain, and the metal-binding site. The crown domain and the metal-binding domain are mammalian-specific and were observed for the first time in the PLAP structure. The crown domain contains a collagen binding loop. A synchrotron radiation x-ray fluorescence study confirms that the metal in the metal-binding site is a calcium ion. Several severe mutations in TNAP occur around this calcium site, suggesting that calcium may be of critical importance for the TNAP function. The presence of this extra metal-binding site gives new insights on the controversial role observed for calcium. PMID- 11395500 TI - The death effector domain-associated factor plays distinct regulatory roles in the nucleus and cytoplasm. AB - Homophilic interactions of death effector domains (DEDs) are crucial for the signaling pathways of death receptor-mediated apoptosis. The machinery that regulates proper oligomerization and autoactivation of procaspase-8 and/or procaspase-10 during T lymphocyte activation determines whether the cells will undergo caspase-mediated apoptosis or proliferation. We screened a yeast two hybrid library by using the DEDs contained in the prodomains of procaspase-8 and procaspase-10 and isolated a DED-associated factor (DEDAF) that interacts with several DED-containing proteins but does not itself contain a DED. DEDAF is highly conserved between human and mouse (98% amino acid identity) and is homologous to a nuclear regulatory protein YAF-2. DEDAF is expressed at the highest levels in lymphoid tissues and placenta. DEDAF interacts with FADD, procaspase-8, and procaspase-10 in the cytosol as well as with the DED-containing DNA-binding protein (DEDD) in the nucleus. At the cell membrane, DEDAF augmented the formation of CD95-FADD-caspase-8 complexes and enhanced death receptor- as well as DED-mediated apoptosis. In the nucleus, DEDAF caused the DEDD protein to relocalize from subnuclear structures to a diffuse distribution in the nucleoplasm. Our data therefore suggest that DEDAF may be involved in the regulation of both cytoplasmic and nuclear events of apoptosis. PMID- 11395501 TI - Targeting of zyxin to sites of actin membrane interaction and to the nucleus. AB - The localization of proteins to particular intracellular compartments often regulates their functions. Zyxin is a LIM protein found prominently at sites of cell adhesion, faintly in leading lamellipodia, and transiently in cell nuclei. Here we have performed a domain analysis to identify regions in zyxin that are responsible for targeting it to different subcellular locations. The N-terminal proline-rich region of zyxin, which harbors binding sites for alpha-actinin and members of the Ena/VASP family, concentrates in lamellipodial extensions and weakly in focal adhesions. The LIM region of zyxin displays robust targeting to focal adhesions. When overexpressed in cells, the LIM region of zyxin causes displacement of endogenous zyxin from focal adhesions. Upon mislocalization of full-length zyxin, at least one member of the Ena/VASP family is also displaced, and the organization of the actin cytoskeleton is perturbed. Zyxin also has the capacity to shuttle between the nucleus and focal adhesion sites. When nuclear export is inhibited, zyxin accumulates in cell nuclei. The nuclear accumulation of zyxin occurs asynchronously with approximately half of the cells exhibiting nuclear localization of zyxin within 2.3 h of initiating leptomycin B treatment. Our results provide insight into the functions of different zyxin domains. PMID- 11395502 TI - Architecture of the human origin recognition complex. AB - All the human homologs of the six subunits of Saccharomyces cerevisiae origin recognition complex have been reported so far. However, not much has been reported on the nature and the characteristics of the human origin recognition complex. In an attempt to purify recombinant human ORC from insect cells infected with baculoviruses expressing HsORC subunits, we found that human ORC2, -3, -4, and -5 form a core complex. HsORC1 and HsORC6 subunits did not enter into this core complex, suggesting that the interaction of these two subunits with the core ORC2-5 complex is extremely labile. We found that the C-terminal region of ORC2 interacts directly with the N-terminal region of ORC3. The C-terminal region of ORC3 was, however, necessary to bring ORC4 and ORC5 into the core complex. A fragment containing the N-terminal 200 residues of ORC3 (ORC3N) competitively inhibited the ORC2-ORC3 interaction. Overexpression of this fragment in U2OS cells blocked the cells in G(1), providing the first evidence that a mammalian ORC subunit is important for the G(1)-S transition in mammalian cells. PMID- 11395503 TI - The Y-box binding protein YB-1 suppresses collagen alpha 1(I) gene transcription via an evolutionarily conserved regulatory element in the proximal promoter. AB - Appropriate expression of collagen type I, a major component of connective tissue matrices, is dependent on tight transcriptional control and a number of trans activating and repressing factors have been characterized. Here we identify the Y box binding protein-1 (YB-1) as a novel repressor of the collagen type alpha1(I) (COL1A1) gene. Collagen type I mRNA and protein levels decreased upon overexpression of YB-1 by transfection in NRK fibroblasts. The human, rat, and mouse COL1A1 promoter -220/+115 contains three putative Y-boxes, one of these sites, designated collagen Y-box element (CYE), includes a Y-box plus an adjacent 3' inverted repeat. DNase-I footprinting and Southwestern blotting with fibroblast nuclear extract demonstrated binding of several nuclear proteins across the CYE, one of which was identified as YB-1. Recombinant YB-1 bound the CYE sequence in gel shift assays with a preference for single-stranded templates. The entire sequence (-88/-48) was required for high affinity binding. Complex formation of endogenous YB-1 with the CYE was established by supershift studies. COL1A1 promoter-reporter constructs were suppressed up to 80% by cotransfection with YB-1 in a variety of cell types. In addition, CYE conferred YB-1 responsiveness on two heterologous promoters further demonstrating the importance of this repressor region. Mung bean nuclease sensitivity analysis suggested that repression is most likely exerted through changes in DNA conformation. PMID- 11395504 TI - Characteristics of a store-operated calcium-permeable channel: sarcoendoplasmic reticulum calcium pump function controls channel gating. AB - We examined the single channel properties and regulation of store-operated calcium channels (SOCC). In human submandibular gland cells, carbachol (CCh) induced flickery channel activity while thapsigargin (Tg) induced burst-like activity, with relatively lower open probability (NP(o)) and longer mean open time. Tg- and CCh-activated channels were permeable to Na(+) and Ba(2+), but not to NMDG, in the absence of Ca(2+). The channels exhibited similar Ca(2+), Na(+), and Ba(2+) conductances and were inhibited by 2-aminoethoxydiphenylborate, xestospongin C, Gd(3+), and La(3+). CCh stimulated flickery activity changed to burst-like activity by (i) addition of Tg, (ii) using Na(+) instead of Ca(2+), (iii) using Ca(2+)-free bath solution, or (iv) buffering [Ca(2+)](i) with BAPTA AM. Buffering [Ca(2+)](i) induced a 2-fold increase in NP(o) of Tg-stimulated SOCC. Reducing free [Ca(2+)] in the endoplasmic reticulum with the divalent cation chelator, N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine (TPEN), induced burst-like channel activity similar to that seen with CCh + Tg. Thus, SOCC is activated by stimulation of muscarinic receptors, inhibition of the sarcoendoplasmic Ca(2+) pump, and lowering [Ca(2+)] in the internal store. Importantly, SOCC activity depends on [Ca(2+)](i) and the free [Ca(2+)] in the internal store. These novel findings reveal that SERCA plays a major role in the gating of SOCC by (i) refilling the internal Ca(2+) store(s) and (ii) decreasing the [Ca(2+)](i)-dependent inhibition. PMID- 11395505 TI - Effects of diabetes insipidus mutations on neurophysin folding and function. AB - Mechanisms underlying the pathogenicity of diabetes insipidus mutations were probed by studying their effects on the properties of bovine oxytocin-related neurophysin. The mutations G17V, DeltaE47, G57S, G57R, and C67STOP were each shown to have structural consequences that would diminish the conformational stability and folding efficiency of the precursors in which they were incorporated, and factors contributing to the origins of these property changes were identified. Effects of the mutations on dimerization of the folded proteins were similarly analyzed. The projected relative impact of the above mutations on precursor folding properties qualitatively parallels the reported relative severity of their effects on the biological handling of the human vasopressin precursor, but quantitative differences between thermodynamic effects and biological impact are noted and explored. The sole mutation for which no clear thermodynamic basis was found for its pathogenicity was 87STOP, suggesting that the region of the precursor deleted by this mutation plays a role in targeting independent from effects on folding, or participates in stabilizing interactions unique to the human vasopressin precursor. PMID- 11395506 TI - Role of the CDC25 homology domain of phospholipase Cepsilon in amplification of Rap1-dependent signaling. AB - Phospholipase Cepsilon (PLCepsilon) is a novel class of phosphoinositide-specific PLC characterized by possession of CDC25 homology and Ras/Rap1-associating domains. We and others have shown that human PLCepsilon is translocated from the cytoplasm to the plasma membrane and activated by direct association with Ras at its Ras/Rap1-associating domain. In addition, translocation to the perinuclear region was induced upon association with Rap1.GTP. However, the function of the CDC25 homology domain remains to be clarified. Here we show that the CDC25 homology domain of PLCepsilon functions as a guanine nucleotide exchange factor for Rap1 but not for any other Ras family GTPases examined including Rap2 and Ha Ras. Consistent with this, coexpression of full-length PLCepsilon or its N terminal fragment carrying the CDC25 homology domain causes an increase of the intracellular level of Rap1.GTP. Concurrently, stimulation of the downstream kinases B-Raf and extracellular signal-regulated kinase is observed, whereas the intracellular level of Ras.GTP and Raf-1 kinase activity are unaffected. In wild type Rap1-overexpressing cells, epidermal growth factor induces translocation of PLCepsilon to the perinuclear compartments such as the Golgi apparatus, which is sustained for at least 20 min. In contrast, PLCepsilon lacking the CDC25 domain translocates to the perinuclear compartments only transiently. Further, the formation of Rap1.GTP upon epidermal growth factor stimulation exhibits a prolonged time course in cells expressing full-length PLCepsilon compared with those expressing PLCepsilon lacking the CDC25 homology domain. These results suggest a pivotal role of the CDC25 homology domain in amplifying Rap1-dependent signal transduction, including the activation of PLCepsilon itself, at specific subcellular locations such as the Golgi apparatus. PMID- 11395507 TI - p65-activated histone acetyltransferase activity is repressed by glucocorticoids: mifepristone fails to recruit HDAC2 to the p65-HAT complex. AB - Glucocorticoids acting through their specific receptor can either enhance or repress gene transcription. Dexamethasone represses interleukin-1beta-stimulated histone acetylation and granulocyte-macrophage colony-stimulating factor expression through a combination of direct inhibition of p65-associated histone acetyltransferase (HAT) activity and by recruiting histone deacetylase 2 (HDAC2) to the p65-HAT complex. Here we show that mifepristone, a glucocorticoid receptor partial agonist, has no ability to induce gene expression but represses interleukin-1beta-stimulated histone acetylation and granulocyte-macrophage colony-stimulating factor release by 50% maximally. Mifepristone was able to inhibit p65-associated HAT activity to the same extent as dexamethasone but failed to inhibit the natural promoter to an equal extent due to an inability to recruit HDAC2 to the p65-associated HAT complex. These data suggest that the maximal repressive actions of glucocorticoids require recruitment of HDAC2 to a p65-HAT complex. These data also suggest that pharmacological manipulation of specific histone acetylation status is a potentially useful approach for the treatment of inflammatory diseases. PMID- 11395508 TI - Defining a link between gap junction communication, proteolysis, and cataract formation. AB - Disruption of the connexin alpha 3 (Cx46) gene (alpha 3 (-/-)) in mice results in severe cataracts within the nuclear portion of the lens. These cataracts are associated with proteolytic processing of the abundant lens protein gamma crystallin, leading to its aggregation and subsequent opacification of the lens. The general cysteine protease inhibitor, E-64, blocked cataract formation and gamma-crystallin cleavage in alpha 3 (-/-) lenses. Using a new class of activity based cysteine protease affinity probes, we identified the calcium-dependent proteases, m-calpain and Lp82, as the primary targets of E-64 in the lens. Profiling changes in protease activities throughout cataractogenesis indicated that Lp82 activity was dramatically increased in alpha 3 (-/-) lenses and correlated both spatially and temporally with cataract formation. Increased Lp82 activity was due to calcium accumulation as a result of increased influx and decreased outflux of calcium ions in alpha 3 (-/-) lenses. These data establish a role for alpha 3 gap junctions in maintaining calcium homeostasis that in turn is required to control activity of the calcium-dependent cysteine protease Lp82, shown here to be a key initiator of the process of cataractogenesis. PMID- 11395509 TI - Conformational changes in four regions of the Escherichia coli ArsA ATPase link ATP hydrolysis to ion translocation. AB - Structures of ArsA with ATP, AMP-PNP, or ADP.AlF(3) bound at the A2 nucleotide binding site were determined. Binding of different nucleotides modifies the coordination sphere of Mg(2+). In particular, the changes elicited by ADP.AlF(3) provide insights into the mechanism of ATP hydrolysis. In-line attack by water onto the gamma-phosphate of ATP would be followed first by formation of a trigonal intermediate and then by breaking of the scissile bond between the beta- and gamma-phosphates. Motions of amino acid side chains at the A2 nucleotide binding site during ATP binding and hydrolysis propagate at a distance, producing conformational changes in four different regions of the protein corresponding to helices H4-H5, helices H9-H10, helices H13-H15, and to the S1-H2-S2 region. These elements are extensions of, respectively, the Switch I and Switch II regions, the A-loop (a small loop near the nucleotide adenine moiety), and the P-loop. Based on the observed conformational changes, it is proposed that ArsA functions as a reciprocating engine that hydrolyzes 2 mol of ATP per each cycle of ion translocation across the membrane. PMID- 11395510 TI - Mechanisms of differential activation of target gene promoters by p53 hinge domain mutants with impaired apoptotic function. AB - Suppression of tumor cell growth by p53 results from the activation of both apoptosis and cell cycle arrest functions that have been shown to be separable activities of p53. We report here that some mutants in the p53 hinge domain, a short linker between the DNA binding and tetramerization domains, differentially activated the promoters of p53 target genes and possessed an impaired apoptotic function. Our results indicate that the hinge domain may play an important role in differentially regulating p53 cell cycle arrest and apoptotic functions. However, the mechanisms by which p53 hinge domain mutants differentially activate its target genes, e.g. p21(WAF1/CIP1) and Bax, remain unknown. To investigate the possible mechanisms, recombinant p21(WAF1/CIP1) and Bax promoters were constructed, resulting in rearrangement of the existing p53 binding sites within a given promoter or actually swapping p53 binding sites between the two promoters. Our results suggest that multiple mechanisms of differential transactivation occur, depending on the molecular nature of the relevant hinge domain mutant, such as the possibility that dual separate DNA binding sites in the p21(WAF1/CIP1) promoter are responsible for the selective transactivation activity of p53 hinge domain mutant del300-327, which has a large deletion in the hinge domain. Lack of ideal p53 binding sites in the Bax promoter results in less potent activation than that seen with the p21(WAF1/CIP1) promoter when it is transactivated by hinge domain point mutant mutR306P or short deletion mutant del300-308 proteins. How the single mutation or the short deletion affect the conformation of p53 and consequently the transactivation of the Bax promoter will require further investigation of the relevant p53 protein: DNA-binding domain by NMR and x-ray crystallographic techniques. PMID- 11395511 TI - Insulin stimulates tyrosine phosphorylation and inactivation of protein-tyrosine phosphatase 1B in vivo. AB - Protein-tyrosine phosphatase (PTP) 1B has been implicated in negative regulation of insulin action, although little is known of the ability of insulin to regulate PTP1B itself. The ability of insulin to regulate phosphorylation and activation of PTP1B was probed in vivo. Challenge with insulin in vivo provoked a transient, sharp increase in the phosphotyrosine content of PTP1B in fat and skeletal muscle that peaked within 15 min. Insulin stimulated a decline of 60--70% in PTP1B activity. In mouse adipocytes, the inhibition of PTP1B activity and increased tyrosine phosphorylation of the enzyme were blocked by the insulin receptor tyrosine kinase inhibitor AG1024. Phosphoserine content of PTP1B declined in response to insulin stimulation. Elevation of intracellular cyclic AMP provokes a sharp increase in PTP1B activity and leads to increased phosphorylation of serine residues and decreased tyrosine phosphorylation. Suppression of cyclic AMP levels or inhibition of protein kinase A leads to a sharp decline in PTP1B activity, a decrease in phosphoserine content, and an increase in PTP1B phosphotyrosine content. PTP1B appears to be a critical point for insulin and catecholamine counter-regulation. PMID- 11395512 TI - Nuclear translocation of cytosolic phospholipase A2 is induced by ATP depletion. AB - Phospholipase A(2) (PLA(2)) enzymes may play a role in cellular injury due to ATP depletion. Renal Madin-Darby canine kidney cells were subjected to ATP depletion to assess the effects of cellular energy metabolism on cytosolic PLA(2) (cPLA(2)) regulation. ATP depletion results in a decrease in soluble cPLA(2) activity and an increase in membrane-associated activity, which is reversed upon restoration of ATP levels by addition of dextrose. In ATP-depleted cells cPLA(2) mass shifts from cytosol to nuclear fractions. GFP-cPLA(2) is localized at the nuclear membrane of stably transfected ATP-depleted LLC-PK(1) cells under conditions where [Ca(2+)](i) is known to increase. cPLA(2) translocation does not occur if the increase in [Ca(2+)](i) increase is inhibited. If [Ca(2+)](i) is allowed to increase when ATP is depleted and the cells are then lysed, cPLA(2) remains associated with nuclear fractions even if the homogenate [Ca(2+)] is markedly reduced. In contrast, cPLA(2), which becomes associated with the nucleus when [Ca(2+)](i) is increased using ionophore, readily dissociates from the nuclear fractions of ATP-replete cells upon reduction of homogenate [Ca(2+)]. Okadaic acid inhibits the ATP depletion-induced association of cPLA(2) with nuclear fractions. Thus energy deprivation results in [Ca(2+)]-induced nuclear translocation, which is partially prevented by a phosphatase inhibitor. PMID- 11395514 TI - Response of an integral granule membrane protein to changes in pH. AB - A key feature of the regulated secretory pathway in neuroendocrine cells is lumenal pH, which decreases between trans-Golgi network and mature secretory granules. Because peptidylglycine alpha-amidating monooxygenase (PAM) is one of the few membrane-spanning proteins concentrated in secretory granules and is a known effector of regulated secretion, we examined its sensitivity to pH. Based on antibody binding experiments, the noncatalytic linker regions between the two enzymatic domains of PAM show pH-dependent conformational changes; these changes occur in the presence or absence of a transmembrane domain. Integral membrane PAM 1 solubilized from rat anterior pituitary or from transfected AtT-20 cells aggregates reversibly at pH 5.5 while retaining enzyme activity. Over 35% of the PAM-1 in anterior pituitary extracts aggregates at pH 5.5, whereas only about 5% aggregates at pH 7.5. PAM-1 recovered from secretory granules and endosomes is highly responsive to low pH-induced aggregation, whereas PAM-1 recovered from a light, intracellular recycling compartment is not. Mutagenesis studies indicate that a transmembrane domain is necessary but not sufficient for low pH-induced aggregation and reveal a short lumenal, juxtamembrane segment that also contributes to pH-dependent aggregation. Taken together, these results demonstrate that several properties of membrane PAM serve as indicators of granule pH in neuroendocrine cells. PMID- 11395513 TI - Stromal inhibition of megakaryocytic differentiation correlates with blockade of signaling by protein kinase C-epsilon and ERK/MAPK. AB - Contact with bone marrow stromal cells maintains normal and leukemic hematopoietic progenitors in an undifferentiated state. Recently, stromal contact has been shown to diminish the yield of megakaryocytes in cultures of primary human hematopoietic stem cells. This inhibition may explain the poor megakaryocytic engraftment frequently observed after bone marrow transplantation. In the current study, stromal co-culture is shown to render K562 cells refractory to megakaryocytic induction. This stromal inhibition correlated with the selective down-regulation in K562 cells of protein kinase C-epsilon (PKC epsilon), which has recently been implicated in regulation of megakaryocytic lineage commitment. In addition, the stromal inhibition correlated with inactivation of the ERK/MAPK pathway, which has also been implicated in promoting megakaryocytic development. Forced expression of PKC-epsilon by retroviral transduction was insufficient to reverse the stromal blockade of ERK/MAPK signaling or of megakaryocytic induction. Thus stromal interruption of ERK/MAPK signaling occurred independently of PKC-epsilon levels and correlated more closely with megakaryocytic blockade. These findings provide potential mechanisms for stromal inhibition of hematopoietic differentiation and possibly for the poor megakaryocytic engraftment seen after bone marrow transplantation. PMID- 11395515 TI - Two consecutive zinc fingers in Sp1 and in MAZ are essential for interactions with cis-elements. AB - The zinc finger proteins Sp1 and Myc-associated zinc finger protein (MAZ) are transcription factors that control the expression of various genes. Regulation of transcription by these factors is based on interactions between GC-rich DNA binding sites (GGGCGG for Sp1 and GGGAGGG for MAZ) and the carboxyl-terminal zinc finger motifs of the two proteins. Sp1 and MAZ have three and six zinc fingers, respectively, and the details of their interactions with cis-elements remain to be clarified. We demonstrate here that Sp1 and MAZ interact with the same GC-rich DNA-binding sites, apparently sharing DNA-binding sites with each other. We found that the DNA binding activities of Sp1 and MAZ depended mainly on consecutive zinc fingers, namely the second and third zinc fingers in Sp1 and the third and fourth zinc fingers in MAZ. Furthermore, the interactions of the zinc finger proteins with the same cis-elements appear to play a critical role in the regulation of gene expression. It seems plausible that two consecutive zinc finger motifs in a zinc finger protein might be essential for interaction of the protein with DNA. PMID- 11395516 TI - Intra-subunit and inter-subunit electron transfer in neuronal nitric-oxide synthase: effect of calmodulin on heterodimer catalysis. AB - In neuronal nitric-oxide synthase (nNOS), calmodulin (CaM) binding is thought to trigger electron transfer from the reductase domain to the heme domain, which is essential for O(2) activation and NO formation. To elucidate the electron transfer mechanism, we characterized a series of heterodimers consisting of one full-length nNOS subunit and one oxygenase-domain subunit. The results support an inter-subunit electron-transfer mechanism for the wild type nNOS, in that electrons for catalysis transfer in a Ca(2+)/CaM-dependent way from the reductase domain of one subunit to the heme of the other subunit, as proposed for inducible NOS. This suggests that the two different isoforms form similar dimeric complexes. In a series of heterodimers containing a Ca(2+)/CaM-insensitive mutant (delta40), electrons transferred from the reductase domain to both hemes in a Ca(2+)/CaM-independent way. Thus, in the delta40 mutant electron transfer from the reductase domains to the heme domains can occur via both inter-subunit and intra-subunit mechanisms. However, NO formation activity was exclusively linked to inter-subunit electron transfer and was observed only in the presence of Ca(2+)/CaM. This suggests that the mechanism of activation of nNOS by CaM is not solely dependent on the activation of electron transfer to the nNOS hemes but may involve additional structural factors linked to the catalytic action of the heme domain. PMID- 11395517 TI - Phenoxybenzamine binding reveals the helical orientation of the third transmembrane domain of adrenergic receptors. AB - Phenoxybenzamine (PB), a classical alpha-adrenergic antagonist, binds irreversibly to the alpha-adrenergic receptors (ARs). Amino acid sequence alignments and the predicted helical arrangement of the seven transmembrane (TM) domains suggested an accessible cysteine residue in transmembrane 3 of the alpha(2)-ARs, in position C(3.36) (in subtypes A, B, and C corresponding to amino acid residue numbers 117/96/135, respectively), as a possible site for the PB interaction. Irreversible binding of PB to recombinant human alpha(2)-ARs (90 nm, 30 min) reduced the ligand binding capacity of alpha(2A)-, alpha(2B)-, and alpha(2C)-AR by 81, 96, and 77%. When the TM3 cysteine, Cys(117), of alpha(2A)-AR was mutated to valine (alpha(2A)-C117V), the receptor became resistant to PB (inactivation, 10%). The beta(2)-AR contains a valine in this position (V(3.36); position number 117) and a cysteine in the preceding position (Cys(116)) and was not inactivated by PB (10 microm, 30 min) (inactivation 26%). The helical orientation of TM3 was tested by exchanging the amino acids at positions 116 and 117 of the alpha(2A)-AR and beta(2)-AR. The alpha(2A)-F116C/C117V mutant was resistant to PB (inactivation, 7%), whereas beta(2)-V117C was irreversibly inactivated (inactivation, 93%), confirming that position 3.36 is exposed to receptor ligands, and position 3.35 is not exposed in the binding pocket. PMID- 11395518 TI - Neural model of the genetic network. AB - Many cell control processes consist of networks of interacting elements that affect the state of each other over time. Such an arrangement resembles the principles of artificial neural networks, in which the state of a particular node depends on the combination of the states of other neurons. The lambda bacteriophage lysis/lysogeny decision circuit can be represented by such a network. It is used here as a model for testing the validity of a neural approach to the analysis of genetic networks. The model considers multigenic regulation including positive and negative feedback. It is used to simulate the dynamics of the lambda phage regulatory system; the results are compared with experimental observation. The comparison proves that the neural network model describes behavior of the system in full agreement with experiments; moreover, it predicts its function in experimentally inaccessible situations and explains the experimental observations. The application of the principles of neural networks to the cell control system leads to conclusions about the stability and redundancy of genetic networks and the cell functionality. Reverse engineering of the biochemical pathways from proteomics and DNA micro array data using the suggested neural network model is discussed. PMID- 11395519 TI - Telomerase can inhibit the recombination-based pathway of telomere maintenance in human cells. AB - Telomere length can be maintained by telomerase or by a recombination-based pathway. Because individual telomeres in cells using the recombination-based pathway of telomere maintenance appear to periodically become extremely short, cells using this pathway to maintain telomeres may be faced with a continuous state of crisis. We expressed telomerase in a human cell line that uses the recombination-based pathway of telomere maintenance to test whether telomerase would prevent telomeres from becoming critically short and examine the effects that this might have on the recombination-based pathway of telomere maintenance. In these cells, telomerase maintains the length of the shortest telomeres. In some cases, the long heterogeneous telomeres are completely lost, and the cells now permanently contain short telomeres after only 40 population doublings. This corresponds to a telomere reduction rate of 500 base pairs/population doubling, a rate that is much faster than expected for normal telomere shortening but is consistent with the rapid telomere deletion events observed in cells using the recombination-based pathway of telomere maintenance (Murnane, J. P., Sabatier, L., Marder, B. A., and Morgan, W. F. (1994) EMBO J. 13, 4953-4962). We also observed reductions in the fraction of cells containing alternative lengthening of telomere-associated promyelocytic leukemia bodies and extrachromosomal telomere repeats; however, no alterations in the rate of sister chromatid exchange were observed. Our results demonstrate that human cells using the recombination-based pathway of telomere maintenance retain factors required for telomerase to maintain telomeres and that once the telomerase-based pathway of telomere length regulation is engaged, recombination-based elongation of telomeres can be functionally inhibited. PMID- 11395520 TI - Phosphatidylserine-specific phospholipase A1 stimulates histamine release from rat peritoneal mast cells through production of 2-acyl-1-lysophosphatidylserine. AB - Lysophosphatidylserine (1-acyl-2-lyso-PS) has been shown to stimulate histamine release from rat peritoneal mast cells (RPMC) triggered by FcepsilonRI (high affinity receptor for IgE) cross-linking, although the precise mechanism of lyso PS production has been obscure. In the present study we show that phosphatidylserine-specific phospholipase A(1), PS-PLA(1), stimulates histamine release from RPMC through production of 2-acyl-1-lyso-PS in the presence of FcepsilonRI cross-linker. The potency of 2-acyl-1-lyso-PS was almost equal to that of 1-acyl-2-lyso-PS. A catalytically inactive PS-PLA(1), in which an active serine residue (Ser(166)) was replaced with an alanine residue did not show such activity. sPLA(2)-IIA, another secretory PLA(2) that is capable of producing lyso PS in vitro, was also a poor histamine inducer against RPMC. PS-PLA(1) significantly stimulated histamine release from crude RPMC, indicating that lyso PS is mainly derived from cells other than mast cells. In agreement with this phenomenon, the enzyme stimulated the histamine release more efficiently when RPMC were mixed with apoptotic Jurkat cells. Under these conditions, lyso-PS with unsaturated fatty acid was released from the apoptotic cells treated with PS PLA(1). Finally, heparin, which has affinity for PS-PLA(1), completely blocked the stimulatory effect of the enzyme. In conclusion, PS-PLA(1) may bind to heparan sulfate proteoglycan, efficiently hydrolyze PS appearing on plasma membranes of apoptotic cells, and stimulate mast cell activation mediated by 2 acyl-1-lyso-PS. PMID- 11395521 TI - Binding of G alpha(o) N terminus is responsible for the voltage-resistant inhibition of alpha(1A) (P/Q-type, Ca(v)2.1) Ca(2+) channels. AB - G-protein-mediated inhibition of presynaptic voltage-dependent Ca(2+) channels is comprised of voltage-dependent and -resistant components. The former is caused by a direct interaction of Ca(2+) channel alpha(1) subunits with G beta gamma, whereas the latter has not been characterized well. Here, we show that the N terminus of G alpha(o) is critical for the interaction with the C terminus of the alpha(1A) channel subunit, and that the binding induces the voltage-resistant inhibition. An alpha(1A) C-terminal peptide, an antiserum raised against G alpha(o) N terminus, and a G alpha(o) N-terminal peptide all attenuated the voltage-resistant inhibition of alpha(1A) currents. Furthermore, the N terminus of G alpha(o) bound to the C terminus of alpha(1A) in vitro, which was prevented either by the alpha(1A) channel C-terminal or G alpha(o) N-terminal peptide. Although the C-terminal domain of the alpha(1B) channel showed similar ability in the binding with G alpha(o) N terminus, the above mentioned treatments were ineffective in the alpha(1B) channel current. These findings demonstrate that the voltage-resistant inhibition of the P/Q-type, alpha(1A) channel is caused by the interaction between the C-terminal domain of Ca(2+) channel alpha(1A) subunit and the N-terminal region of G alpha(o). PMID- 11395522 TI - Functional groups required for the stability of yeast RNA triphosphatase in vitro and in vivo. AB - Cet1, the RNA triphosphatase component of the yeast mRNA capping apparatus, catalyzes metal-dependent gamma-phosphate hydrolysis within the hydrophilic interior of an eight-strand beta barrel (the "triphosphate tunnel"), which rests upon a globular protein core (the "pedestal"). We performed a structure-guided alanine scan of 17 residues located in the tunnel (Ser(373), Thr(375), Gln(405), His(411), Ser(429), Glu(488), Thr(490)), on the tunnel's outer surface (Ser(378), Ser(487), Thr(489), His(491)), at the tunnel-pedestal interface (Ile(304), Met(308)) and in the pedestal (Asp(315), Lys(317), Arg(321), Asp(425)). Alanine mutations at 14 positions had no significant effect on Cet1 phosphohydrolase activity in vitro and had no effect on Cet1 function in vivo. Two of the mutations (R321A and D425A) elicited a thermosensitive (ts) yeast growth phenotype. The R321A and D425A proteins had full phosphohydrolase activity in vitro, but were profoundly thermolabile. Arg(321) and Asp(425) interact to form a salt bridge within the pedestal that tethers two of the strands of the tunnel. Mutations R321Q and D411N resulted in ts defects in vivo and in vitro, as did the double-mutant R321A-D435A, whereas the R321K protein was fully stable in vivo and in vitro. These results highlight the critical role of the buried salt bridge in Cet1 stability. Replacement of Ser(429) by alanine or valine elicited a cold sensitive (cs) yeast growth phenotype. The S429A and S429V proteins were fully active when produced in bacteria at 37 degrees C, but were inactive when produced at 17 degrees C. Replacement of Ser(429) by threonine partially suppressed the cold sensitivity of the Cet1 phosphohydrolase, but did not suppress the cs growth defect in yeast. PMID- 11395524 TI - Mandibular advancement using an intra-oral osteogenic distraction technique: a report of three clinical cases. AB - Osteogenic distraction has been used for decades to lengthen limbs and now attention is focused upon its use within the craniofacial skeleton. This paper addresses distraction of the mandible. It is proposed that mandibular osteogenic distraction could be a possible adjunct to the orthodontic treatment of those adult patients with skeletal anomalies, who would benefit from combined orthodontic/orthognathic treatment. Three consecutive cases from one unit are presented, where adult patients with severe Class II division 1 malocclusions have undergone orthodontic treatment combined with mandibular osteogenic distraction, instead of conventional bilateral sagittal split osteotomies. PMID- 11395523 TI - Phosphorylation of the yeast phospholipid synthesis regulatory protein Opi1p by protein kinase C. AB - Opi1p is a negative regulator of expression of phospholipid-synthesizing enzymes in the yeast Saccharomyces cerevisiae. In this work, we examined the phosphorylation of Opi1p by protein kinase C. Using a purified maltose-binding protein-Opi1p fusion protein as a substrate, protein kinase C activity was time- and dose-dependent, and dependent on the concentrations of Opi1p and ATP. Protein kinase C phosphorylated Opi1p on a serine residue. The Opi1p synthetic peptide GVLKQSCRQK, which contained a protein kinase C sequence motif at Ser(26), was a substrate for protein kinase C. Phosphorylation of a purified S26A mutant maltose binding protein-Opi1p fusion protein by the kinase was reduced when compared with the wild-type protein. A major phosphopeptide present in purified wild-type Opi1p was absent from the purified S26A mutant protein. In vivo labeling experiments showed that the phosphorylation of Opi1p was physiologically relevant, and that the extent of phosphorylation of the S26A mutant protein was reduced by 50% when compared with the wild-type protein. The physiological consequence of the phosphorylation of Opi1p at Ser(26) was examined by measuring the effect of the S26A mutation on the expression of the phospholipid synthesis gene INO1. The beta galactosidase activity driven by an INO1-CYC-lacI'Z reporter gene in opi1Delta mutant cells expressing the S26A mutant Opi1p was about 50% lower than that of cells expressing the wild-type Opi1p protein. These data supported the conclusion that phosphorylation of Opi1p at Ser(26) mediated the attenuation of the negative regulatory function of Opi1p on the expression of the INO1 gene. PMID- 11395525 TI - Ameloblastic fibroma of the anterior maxilla presenting as a complication of tooth eruption: a case report. AB - Ameloblastic fibroma is a rare mixed odontogenic tumour, which is extremely uncommon in the anterior maxillary region. A case report is presented where failure of eruption of an upper central incisor was the presenting feature. PMID- 11395526 TI - Morphology of the temporomandibular joint in skeletal class iii symmetrical and asymmetrical cases: a study by cephalometric laminography. AB - The aetiology of asymmetric growth in the mandible is not well understood. Previous studies have indicated that the functional lateral shift of the mandible in the period of prepubertal growth may translate to a true skeletal asymmetry, exclusively in skeletal Class III malocclusion. This asymmetry develops more characteristic features during the pubertal and post-pubertal growth periods. Early correction of a functional lateral shift of the mandible is recommended. The purpose of this study was to examine the relationship between the morphology of the temporomandibular joints and asymmetry in skeletal Class III malocclusion in adult female patients. Cephalometric and laminographic findings in 36 asymmetric skeletal Class III patients with a lateral shift of mandible (group 3) were compared to those of 25 symmetric skeletal Class I patients (group 1) and the same number of symmetric skeletal Class III malocclusions (group 2). All the patients had received no orthodontic treatment. The results showed that the TMJ of the side to which the mandible shifted showed a significantly narrower and shorter shape of the condyle head, smaller superior condylar space, and steeper eminence than those of the unshifted side. PMID- 11395527 TI - Rapid palatal expansion in mixed dentition using a modified expander: a cephalometric investigation. AB - The aims of this investigation were to cephalometrically study the short-term skeletal and dental modifications induced by rapid palatal expansion in a sample of 20 patients (10 male, 10 female), aged 6-10 years (mean age 8 years) in mixed dentition with a uni- or bilateral posterior crossbite, a mild skeletal Class II malocclusion, and an increased vertical dimension (FMA, SN/?GoGn), and to compare them with an untreated matched control group of 20 subjects (10 male and 10 female), mean age 8 years. Cephalometric analysis showed that the maxilla displayed a tendency to rotate downwards and backward, resulting in a statistically significant increase of the SN/?PP angle (T0 = 9*95 degrees, T1 = 11*60 degrees, P < 0*01) and the SN-ANS linear value (T0 = 49*50 mm, T1 = 51*10 mm, P < 0*05). In addition, there was a statistically significant alteration of the anterior total facial height N-Me (T0 = 113*15mm, T1 = 114*15 mm, P < 0*05) and for the dental upper molar measurement U6-PP (T0 = 19*70 mm, T1 = 20*30 mm, P < 0*05). The small alterations found in the anterior total facial height and in the sagittal angles agree with previous studies, and suggest that RPE can be also used in subjects with a tendency to vertical growth and a skeletal Class II malocclusion. PMID- 11395528 TI - A validation of two orthognathic model surgery techniques. AB - In order to create an evidence-based orthognathic surgery planning protocol, an investigation of two popular model surgery techniques, the Lockwood keyspacer and the Eastman anatomically-orientated system was carried. This determined (a) the accuracy of positioning of the maxillary cast according to the prescribed treatment plan and (b) the relocation of the maxilla after a simulated Le Fort I down fracture osteotomy using the intermediate wafer as a guide. Fifteen patients five Class II division 1, five Class II division 2, and five Class III-were included in the study. All the measurements were taken with Erickson's vertically mounted electronic caliper and variations from the treatment plan were analysed. The mean model surgery positioning errors +/- SD (mm) were: (i) vertical plane Lockwood -0*8 +/- 1*6 and Eastman 0*00 +/- 1*0 (P = 0*0001); (ii) anteroposterior plane-Lockwood 1*2 +/- 1*8 and Eastman -0*1 +/- 1*4 (P = 0*05); and (iii) transverse plane-Lockwood 0*9 +/- 0*9 and Eastman 1*0 +/- 0*9 (P = 0*34). After the simulated osteotomy, the mean errors +/- SD were: (i) vertical plane-Lockwood -0*5 +/- 1*5 and Eastman 0*3 +/- 1*1 (P = 0*001); (ii) in anteroposterior plane Lockwood 0*8 +/- 2*0 and Eastman 0*7 +/- 1*0 (P = 0*89); and (iii) transverse plane-Lockwood 0*8 +/- 0*6 and Eastman 0*7 +/- 0*5 (P = 0*83). The Eastman technique was relatively better especially in the vertical plane. The variations from the treatment plan were on the whole anatomically small, but in some cases could be clinically significant. PMID- 11395530 TI - Peer review. PMID- 11395529 TI - An accurate three-dimensional cephalometric system: a solution for the correction of cephalic malpositioning. AB - In recent years, methods have been developed that calculate three-dimensional (3D) co-ordinates of orthodontic landmarks from lateral and frontal cephalograms. However, precise measurement has been impossible with these methods because, although they corrected the magnification of the image, they did not correct 3D cephalic malpositioning that occurs during the measurement of human subjects. In this study, we developed a 3D cephalometric system that corrected not only for magnification of the image, but also 3D cephalic malpositioning during cephalogram exposure. Magnification of the image was corrected for first. Cephalic revolution was then sequentially corrected and divided into elements of x-, y-, and z-axes. The origin was parallelly translated to the mid-point of bilateral porion. In order to examine the accuracy of this system, seven human dry skulls were measured. The accuracy unaffected by the cephalic revolution in any direction and standard errors was within 0.8 mm in any orthodontic landmarks. It was suggested that this measurement system would have sufficient accuracy for clinical application. The results indicated that precise cephalometric measurement was possible with this system and it was suggested that its clinical application would be possible. PMID- 11395531 TI - Quality of life and its importance in orthodontics. AB - Over the last 10-15 years, the terms quality of life (QOL) and health-related quality of life (HRQL) have been seen increasingly in medical literature. Much of the orthodontic treatment that is undertaken is justified on the basis of improving health-related quality of life. With this in mind, studying HRQL in orthodontic patients has the potential to provide information about treatment needs and outcomes, and may also facilitate improved care. Clinicians should therefore be aware of some of the ways in which health-related quality of life may be assessed. The first part of this review article looks at the general concepts of health-related quality of life, whilst the second section focuses on dentistry and orthodontics. PMID- 11395532 TI - Background considerations to facial aesthetics. AB - This paper examines the nature of our objectives in attempting to improve facial appearance. Questions are addressed concerning the basis for any collective agreement on "improvement" and the nature of an agreed "ideal", including an exploration of the reasons for its existence. The discussion focuses on the concept of "averageness", as well as supplementary hypotheses. Finally, the origins and validity of contemporary clinical guidelines are addressed. PMID- 11395533 TI - Statistical applications in orthodontics. Part III. How large a study is needed? PMID- 11395534 TI - Functional occlusion: II. The role of articulators in orthodontics. AB - Opinion is divided on whether there is a place for the use of semi-adjustable dental articulators in orthodontics. In this review we explore the validity and reproducibility of the techniques involved in mounting study models on a semi adjustable dental articulator. We also look at the role of articulated study models in orthodontic diagnosis and treatment planning, in the finishing stages of orthodontics and in planning for orthognathic surgery. We report that each of the many stages involved in mounting study models on a semi-adjustable articulator is a potential source of error and that only if the technique is carried out with a high degree of accuracy is it worth the additional chairside time. PMID- 11395536 TI - Rationalizing retention regimes. PMID- 11395535 TI - Orthodontic hyper knowledge--a review. PMID- 11395540 TI - Challenges of comparative expression profiling studies of complex diseases: mouse models of myocardial hypertrophy. Focus on "Divergent transcriptional responses to independent genetic causes of cardiac hypertrophy". PMID- 11395541 TI - Further chromosomal mapping of a blood pressure QTL in Dahl rats on chromosome 2 using congenic strains. AB - Both linkage and use of congenic strains have shown that a region on rat chromosome 2 (Chr 2) of Dahl salt-sensitive rats (S) contained a quantitative trait locus (QTL) for blood pressure (BP). A congenic strain was made by replacing a segment of the S rat by the homologous region of the Milan normotensive (MNS) rat. Since the region was roughly 80 cM in size, a further reduction is required toward the final identification of the QTL. Currently, three congenic substrains were made by replacing smaller sections within the 80 cM. Each strain contains a specific region of MNS in the S genetic background. Two of the three congenic strains shared a segment in common, and both showed a BP-lowering effect. One of the three congenic strains carried a unique segment and had the same BP as S. Deducing the fragment shared in the two substrains having an effect, the BP QTL has to be present in a region of roughly 15 cM. In contrast to BP, heart rates of all the congenic rats were the same as that of the S rat. Thus BP and the heart rate are under the control of independent genetic determinants. PMID- 11395542 TI - Mapping of microsatellite loci and association of aorta atherosclerosis with LG VI markers in the rabbit. AB - Twenty-three rabbit microsatellites were extracted from the EMBL nucleotide database. Nine of these markers, together with nine earlier published microsatellite markers, were found to be polymorphic between the AX/JU and IIIVO/JU inbred strains. By using an F(2) intercross we could integrate five markers into the rabbit linkage map. One anonymous microsatellite marker could be assigned to chromosome 1, and one microsatellite marker, located within the metallothionein-1 gene, could be added to linkage group VI (LG VI). Three microsatellite markers (one anonymous, one located within the PMP2 gene, and one located within the FABP6 gene) constitute a new linkage group (LG XI). We also measured the degree of dietary cholesterol-induced aorta atherosclerosis in the F(2) animals. A significant cosegregation was found between the degree of aorta atherosclerosis and the allelic variation of the biochemical marker Est-2 on LG VI in male rabbits. This association was not found in female rabbits. PMID- 11395543 TI - Divergent transcriptional responses to independent genetic causes of cardiac hypertrophy. AB - To define molecular mechanisms of cardiac hypertrophy, genes whose expression was perturbed by any of four different transgenic mouse hypertrophy models [protein kinase C-epsilon activation peptide (PsiepsilonRACK), calsequestrin (CSQ), calcineurin (CN), and Galpha(q)] were compared by DNA microarray analyses using the approximately 8,800 genes present on the Incyte mouse GEM1. The total numbers of regulated genes (tens to hundreds) correlated with phenotypic severity of the model (Galpha(q) > CN > CSQ > PsiepsilonRACK), but demonstrated that no single gene was consistently upregulated. Of the three models exhibiting pathological hypertrophy, only atrial natriuretic peptide was consistently upregulated, suggesting that transcriptional alterations are highly specific to individual genetic causes of hypertrophy. However, hierarchical-tree and K-means clustering analyses revealed that subsets of the upregulated genes did exhibit coordinate regulatory patterns that were unique or overlapping across the different hypertrophy models. One striking set consisted of apoptotic genes uniquely regulated in the apoptosis-prone Galpha(q) model. Thus, rather than identifying a single common hypertrophic cardiomyopathy gene program, these data suggest that extensive groups of genes may be useful for the prediction of specific underlying genetic determinants and condition-specific therapeutic approaches. PMID- 11395544 TI - Ureteric bud derivatives express angiotensinogen and AT1 receptors. AB - Inactivation of the renin-angiotensin system interferes with the morphogenesis of the renal medulla. Thus ureteric bud (UB) derivatives may be a target for angiotensin production and action. To begin to test this hypothesis, we examined the cellular expression of angiotensinogen (Ao) and AT(1) receptor proteins during rat metanephrogenesis by immunohistochemistry. In addition, we tested whether UB-derived cells in culture express the Ao and AT(1) proteins. On embryonic day E15, Ao and AT(1) are expressed in the UB branches and stromal mesenchyme. S-shaped bodies, including the vascular cleft, express AT(1) but not Ao. The metanephric mesenchyme and pretubular aggregates are Ao negative and AT(1) negative. Expression of Ao and AT(1) in UB branches and ampullae is also observed on E16. However, UB expression of Ao is transient and is no longer detectable in the developing distal nephron beyond E17. On E17, both Ao and AT(1) are expressed in capillary loop glomeruli and proximal tubules, whereas UB branches express AT(1) only. By E18, the majority of Ao immunoreactivity is clustered in terminally differentiated proximal tubules, whereas AT(1) receptors are expressed in both proximal and distal nephron segments. The specificity of Ao and AT(1) staining was documented by the elimination/attenuation of immunoreactivity after preadsorption of the primary antibodies with their respective antigens. Consistent with the in vivo findings, the AT(1) protein is abundantly expressed in cellular lysates of mouse UB (E11.5) and IMCD3 (adult) cells. Moreover, AT(1) receptors in UB and IMCD3 cells are functional, since angiotensin II treatment elicits the tyrosine phosphorylation of the mitogen activated protein kinases, ERK1/2. To our knowledge, this is the first demonstration of Ao and AT(1) protein expression in the developing distal nephron. Angiotensin II may have a paracrine role in the ontogeny of the collecting system. PMID- 11395545 TI - Contrasting pharmacological ETB receptor blockade with genetic ETB deficiency in renal responses to big ET-1. AB - Renal clearance studies were conducted to determine the role of ET(B) receptors in the renal response to big endothelin-1 (big ET-1). Two series of experiments were conducted on Inactin-anesthetized rats to contrast acute pharmacological blockade of ET(B) receptors vs. genetic ET(B) receptor deficiency. In the first series, Sprague-Dawley rats were given either ET(B)-selective antagonist, A 192621, or vehicle (0.9% NaCl) prior to infusion of big ET-1 (10 pmol.kg(-1).min( 1)) for 60 min. A-192621 significantly increased baseline mean arterial pressure (MAP; 102 +/- 4 vs. 141 +/- 6 mmHg, P < 0.05) and urine flow rate (0.5 +/- 0.1 vs. 1.3 +/- 0.2 microl/min, P < 0.05) without any effect on glomerular filtration rate (GFR) or effective renal plasma flow (ERPF). Big ET-1 significantly increased MAP in both groups but to a higher level in rats given antagonist (120 +/- 6 vs. 169 +/- 6 mmHg, P < 0.05). Big ET-1 increased urine flow in control rats but decreased in rats given antagonist. GFR and ERPF were decreased in rats given big ET-1, an effect that was exaggerated by ET(B) blockade. Another series of experiments examined the response to big ET-1 in rats lacking functional renal ET(B) receptors, known as spotting lethal (sl) rats. Surprisingly, rats heterozygous (sl/+) for ET(B) receptor deficiency had a significantly higher baseline MAP compared with homozygous (sl/sl) rats (134 +/- 6 vs. 112 +/- 7 mmHg, P < 0.05), although other variables were similar. Big ET-1 produced no significant change in MAP in either group. Urine flow, GFR, and ERPF were significantly decreased in both groups, although these changes were much larger in sl/sl rats. These experiments indicate that the ET(B) receptor plays an important role in limiting the renal hemodynamic response to big ET-1. Furthermore, the diuretic actions of big ET-1 require a functional ET(B) receptor. PMID- 11395546 TI - Ren1d and Ren2 cooperate to preserve homeostasis: evidence from mice expressing GFP in place of Ren1d. AB - To distinguish the contributions of Ren1(d) and Ren2 to kidney development and blood pressure homeostasis, we placed green fluorescent protein (GFP) under control of the Ren1(d) renin locus by homologous recombination in mice. Homozygous Ren1(d)-GFP animals make GFP mRNA in place of Ren1(d) mRNA in the kidney and maintain Ren2 synthesis in the juxtaglomerular (JG) cells. GFP expression provides an accurate marker of Ren1(d) expression during development. Kidneys from homozygous animals are histologically normal, although with fewer secretory granules in the JG cells. Blood pressure and circulating renin are reduced in Ren1(d)-GFP homozygotes. Acute administration of losartan decreases blood pressure further, suggesting a role for Ren2 protein in blood pressure homeostasis. These studies demonstrate that, in the absence of Ren1(d), Ren2 preserves normal kidney development and prevents severe hypotension. Chronic losartan treatment results in compensation via recruitment of both Ren1(d)- and Ren2-expressing cells along the preglomerular vessels. This response is achieved by metaplastic transformation of arteriolar smooth muscle cells, a major mechanism to control renin bioavailability and blood pressure homeostasis. PMID- 11395547 TI - Sclerosing epithelioid fibrosarcoma: a study of 16 cases and confirmation of a clinicopathologically distinct tumor. AB - Sclerosing epithelioid fibrosarcoma (SEF) is an uncommon tumor of deep soft tissues, originally described in 1995 by Meis-Kindblom et al. In the current study, the authors identified 16 cases of SEF in the pathology files of their institutions and studied their pathologic features and disease course. The group consisted of six male and 10 female patients (age range, 14-55 years; mean age, 40 years), and the tumors were located in a limb or limb girdle (n = 7), base of the penis (n = 1), back or chest wall (n = 3), and head and neck (n = 5). Tumor size ranged from 3.7 to 22 cm (mean, 8.9 cm). Histologically, the SEFs were composed predominantly of small to moderate-size round to ovoid, relatively uniform cells, often with clear cytoplasm, embedded in a hyalinized fibrous stroma. The only consistent immunohistochemical finding was a strong, diffuse reactivity of tumor cells for vimentin. Ultrastructural analysis performed in eight cases confirmed their fibroblastic nature. Bone invasion and tumor necrosis, features not reported before, were found in six cases each. Treatment consisted of intralesional excision (n = 2), attempted wide local excision (n = 11), and amputation (n = 3), with either adjuvant radiation therapy (n = 9) or chemotherapy (n = 3). Follow-up of at least 1 year in 14 cases revealed persistent disease or local recurrence in seven patients (50%), and distant metastasis in 12 patients (86%). Eight patients (57%) died of disease 16 to 86 months after diagnosis. Five patients were alive with disease as of last follow up. SEF shares some pathologic features with two other fibrosing fibrosarcomas, low-grade fibromyxoid sarcoma and hyalinizing spindle cell tumor with giant rosettes, but in the authors' experience behaves clinically as a fully malignant sarcoma. PMID- 11395548 TI - Eccrine porocarcinoma (malignant eccrine poroma): a clinicopathologic study of 69 cases. AB - The clinicopathologic characteristics of 69 cases of eccrine porocarcinoma (EP) have been studied. Seven cases of purely in situ disease are included. Forty patients were female, 29 male with ages ranging from 29 to 91 years (mean 73 years). The lower extremity represented the single most common site (44%). Other common sites were the trunk (15 cases, 24%) and head (11 cases, 18%). The histologic diagnosis of EP was predicated on the basis of an irregular tumor at least partly formed of characteristic poromatous basaloid epithelial cells displaying ductal differentiation, and significant cytologic atypia. Forty-seven tumors (68%) contained mature well-formed eccrine ducts having an eosinophilic luminal cuticle, with the remaining tumors containing small ill-formed ducts and/or intracytoplasmic lumina. All ducts were discernible via light microscopy and in 49 cases were highlighted with DPAS stain and/or CEA/EMA immunocytochemistry. A variant with a broad pushing tumor margin and marked nuclear pleomorphism showed some resemblance to proliferative bowenoid dysplasia. In 11 cases (18%) the tumors appeared to arise in continuity with a benign preexistent poroma. A variety of histologic patterns were displayed including clear, squamous, and spindle cell differentiation, mucus cell metaplasia, and colonization by melanocytes. Lymphovascular invasion was present in 9 cases (15%). Three cases showed pagetoid extension of malignant cells (epidermotropism) and appeared to be multifocal. Follow-up was available in 54 patients (78%) with 9 (17%) experiencing local recurrence, 10 developing lymph node metastases (19%), and 6 (11%) experiencing distant metastases or death. Mitoses, the presence of lymphovascular invasion, and tumor depth >7 mm were associated with a poorer prognosis. Dividing tumors into those with a "pushing" or "infiltrating" advancing margin was also predictive of outcome with the latter having an increased risk of local recurrence. This report, the largest series of EP to date, suggests that the incidence of aggressive behavior is less than popularly believed. Furthermore, EP can display a wide variety of histologic patterns that may lead to diagnostic error in the unwary. The large number of cases in this series enables a reliable evaluation of prognostic parameters. A more aggressive clinical course may be indicated by more than 14 mitoses per high power field (hazard ratio [HR] for death 17.0, 95% confidence interval [CI] 2.71-107), lymphovascular invasion by tumor (HR 4.41, CI 1.13-17.2), and depth >7 mm (HR 5.49, CI 1.0-30.3). Thus, mitoses, lymphovascular invasion, and tumor depth should be evaluated in these tumors. We also suggest that tumors presenting an "infiltrative" advancing margin are particularly prone to local recurrence and require wide excision with close attention to the surgical margins by the reporting pathologist. PMID- 11395549 TI - Inflammatory pseudotumor-like follicular dendritic cell tumor: a distinctive low grade malignant intra-abdominal neoplasm with consistent Epstein-Barr virus association. AB - Follicular dendritic cell (FDC) tumors are uncommon neoplasms that can involve lymph nodes or extranodal sites. They can exhibit a broad spectrum of histologic appearances and behavior, but the intra-abdominal ones usually pursue an aggressive course. The purpose of this study was to characterize a distinctive variant of FDC tumor morphologically mimicking inflammatory pseudotumor through analysis of the clinicopathologic features of 11 cases. The patients included 10 women and one man (age range, 19-61 years; median age, 40 years) who presented with abdominal discomfort or pain. Six patients had systemic symptoms such as marked weight loss, fever, or malaise. All tumors occurred in intra-abdominal sites: liver (n = 7), spleen (n = 3), and peripancreatic region (n = 1). Of the nine patients with follow-up data, six were alive and well, one developed recurrence at 9 years, and two had repeated recurrences over many years. Grossly, the tumors were usually solitary and fleshy, punctuated by areas of hemorrhage and necrosis. Histologically, in a background of abundant lymphocytes and plasma cells were dispersed spindle or ovoid cells with vesicular nuclei and distinct nucleoli. The degree of nuclear atypia was variable, and some nuclei could be grotesque or resemble Reed-Sternberg cells. Focally, spindle cell fascicles could be formed. The atypical cells were immunoreactive for FDC markers such as CD21/CD35, CD23, and CNA.42. In situ hybridization for Epstein-Barr virus (EBV) encoded RNA was positive in all cases, remarkably highlighting the spindle cells and their atypia. EBV-latent membrane protein-1 was expressed commonly, albeit often focally and weakly. Therefore, inflammatory pseudotumor-like FDC tumor represents a distinctive variant of FDC tumor that differs from conventional FDC tumor in the following aspects: marked female predominance; selective localization in intra-abdominal sites, especially the liver and spleen; frequent presence of systemic symptoms; indolent behavior despite an intra-abdominal location; dispersed distribution of tumor cells and prominent lymphoplasmacytic infiltration; and consistent association with EBV. PMID- 11395550 TI - Cutaneous b-cell lymphomas of follicular and marginal zone types: use of Bcl-6, CD10, Bcl-2, and CD21 in differential diagnosis and classification. AB - Cutaneous follicular lymphomas (FLs) and cutaneous B-cell lymphomas of extranodal marginal zone (MZL)/mucosal-associated lymphoid tissue (MALT) type may have morphologic overlap, despite the fact that they are thought to be of distinct derivation (germinal center vs. postgerminal center). The problem is compounded by the reported absence of bcl-2 expression by many cutaneous FLs, leading to speculation that cutaneous FL may be unrelated to nodal FL. The authors analyzed the expression of the germinal center-associated antigens bcl-6 and CD10 and of bcl-2 in 18 cutaneous B-cell lymphomas (10 FLs and eight MZLs), in relationship to CD21+ follicular structures, to clarify the relationship of nodal to cutaneous FLs and to explore the value of these antigens in differential diagnosis. The authors studied 10 cutaneous FLs (seven primary and three secondary) and eight MZLs (six primary and two secondary). The FLs (found in six men and four women age 45-75 years) involved the trunk (n = 3) and scalp, face and neck (n = 7). The MZLs (found in five women and three men age 34-81 years) involved the trunk (n = 4), face and neck (n = 2), and arm (n = 2). Immunostaining for CD21, bcl-6, CD10, and bcl-2 allowed the delineation of compartments within the tumors and yielded distinct patterns of staining in FL and MZL. In both follicular and interfollicular/diffuse areas of FL the neoplastic cells were bcl-6+ (10 of 10), often CD10+ (seven of 10, four of seven primary), and bcl-2+ (nine of 10, six of seven primary). Only three of seven cases (one of five primary) had bcl-2 rearrangement detectable by polymerase chain reaction. In the MZLs, the neoplastic B-cells were bcl-6-, CD10-, and bcl-2+ (eight of eight). Three patterns of CD21+ follicles were identified in MZL: reactive germinal centers, uniformly bcl-6+, CD10+, and bcl-2- (five of eight MZLs); colonized follicles, both bcl-6-, bcl-2+, and L26+ cells, and bcl-6+ and bcl-2- cells (five of eight MZLs); and expanded/colonized follicular dendritic cell meshworks, bcl-6- and bcl 2+ B cells with rare residual bcl-6+ and bcl-2- cells (four of eight MZLs). The authors conclude that cutaneous FLs express bcl-6 uniformly, usually express CD10 and bcl-2, and have a follicular pattern similar to nodal FL and consistent with a germinal center origin. The immunophenotype of cutaneous FL is distinct from that of cutaneous MZL, which is negative for bcl-6 and CD10. Colonized follicles in MZL, identified by CD21+ follicular dendritic cell meshworks, contained numerous bcl-6- and bcl-2+ B cells, and were readily distinguished from neoplastic follicles in FL. Conversely, CD21- interfollicular and diffuse areas in FLs contained bcl-6+ and CD10+ cells, which were not seen in diffuse areas of MZLs. Thus, the combination of bcl-2, bcl-6, and CD21 staining is useful for the distinction of cutaneous MZL from cutaneous FL. PMID- 11395551 TI - Is lymphoplasmacytic lymphoma/immunocytoma a distinct entity? A clinicopathologic study of 20 cases. AB - Lymphoplasmacytic lymphoma/immunocytoma (LLI) was defined initially as a small B cell lymphoma with plasmacytoid or plasmacytic features. Because other types of small B-cell lymphoma, particularly marginal zone B-cell lymphoma may exhibit plasmacytic differentiation, the revised European-American lymphoma classification and World Health Organization has defined LLI more narrowly to exclude other small B-cell lymphomas. The goal of this study was to reevaluate LLI as a clinicopathologic entity. Twenty cases were selected from 43 previously diagnosed as "small lymphocytic lymphoma, plasmacytoid" or "immunocytoma" from 1985 to 1998. Cases fulfilling the criteria for B-cell small lymphocytic lymphoma, follicular lymphoma, marginal zone B-cell lymphoma, or other types of B cell lymphoma were excluded. The histopathology and immunoreactivity for CD20, CD79a, CD3, CD43, CD23, CD5, kappa, lambda, and immunoglobulins (Ig's) M, G, and A were reviewed, in addition to available clinical findings. There were 13 men and seven women, with a mean age of 69 years. Five patients had documented Waldenstrom's macroglobulinemia (WM). Three architectural patterns were observed. Pattern A (seven of 20) showed open sinuses, small follicles, and hemosiderosis; pattern B (four of 20) showed hyperplastic follicles; and pattern C (nine of 20) showed diffuse effacement. Epithelioid histiocytes were prominent in patterns B and C but absent in A. Cytologically, six of 20 were polymorphous with 10% to 40% transformed cells; 14 of 20 were lymphoplasmacytic. Five cases showed minor foci of monocytoid B cells. One case showed a composite histology of LLI and small lymphocytic lymphoma. Amyloid was present in two cases. All cases were CD20 and/or CD79a immunoreactive, with two of 20 positive for CD43. Twelve cases were kappa monoclonal and eight cases were lambda monoclonal. Twelve of 17 cases that could be evaluated were positive for IgM and five were positive for IgG. All cases were negative for CD5 and CD23 with the exception of the one case with a composite histology. Eleven of 20 patients with available follow-up died of disease (median, 48 months), and eight of 20 are alive with disease at a follow up of 6 months to 2 years. LLI does appear to represent a distinct clinicopathologic entity even though it shows morphologic heterogeneity and overlapping features with marginal zone B-cell lymphoma and small lymphocytic lymphoma. Recognition of LLI is important because the overall prognosis may be worse than for other types of small B-cell lymphomas. PMID- 11395552 TI - Cystic endocrine tumors of the pancreas: clinical, radiologic, and histopathologic features in 13 cases. AB - Cystic endocrine tumors of the pancreas are rare and raise difficult clinical problems. Our aims were to reevaluate the diagnostic and therapeutic strategy and to assess their histopathologic characteristics. Thirteen cystic endocrine tumors diagnosed in 10 patients were included. Clinical, radiologic, and pathologic data were reviewed. There were 6 male and 4 female patients (median age, 46 yrs). Six patients had evidence of multiple endocrine neoplasia type 1 (MEN1) disease. Four had a functional endocrine syndrome. Ten tumors were visible on imaging studies. The most suggestive radiologic features were the existence of a peripheral hypervascular rim (10 cases) and images of cyst into cyst (two cases). On gross and histologic examinations, two distinct types were present. Macrocystic tumors (six cases) were unilocular and limited by a thick wall containing nests of tumor cells. Microcystic tumors (seven cases) were characterized by the presence of multiple cystic spaces directly lined by tumor cells. Surgical resection was performed in all cases. Three patients had lymph node metastases at the time of diagnosis. One patient is dead with metastatic dissemination. The others are alive without recurrence or metastasis. The diagnosis of endocrine tumor must be considered for any pancreatic cyst discovered in a patient with a history of MEN1 syndrome or with clinical features suggestive of this syndrome. Cystic pancreatic endocrine tumors must be treated by surgical resection because of their possible malignant evolution. PMID- 11395553 TI - Anaplastic lymphoma kinase expression in inflammatory pseudotumors. AB - Anaplastic lymphoma kinase (ALK), a hallmark of anaplastic large cell lymphoma, has recently been implicated in the genesis of some inflammatory pseudotumors (inflammatory myofibroblastic tumors) in children and young adults. The aim of this study was to determine the frequency of its expression among inflammatory pseudotumors, and to characterize the clinicopathologic features of the positive cases. Sixty-one cases of inflammatory pseudotumors were retrieved from the surgical pathology archives and consultation files. Paraffin sections were immunostained with the antibody ALK1. The patients ranged in age from 0.5 to 79 years (median age, 50 years), with 10 patients (16.4%) younger than 20 years. Five cases (8.2%) were ALK+, including two of six urogenital inflammatory myofibroblastic tumors, none of eight pulmonary inflammatory pseudotumors, three (one adrenal, one small bowel, one liver) of 31 extrapulmonary inflammatory pseudotumors, none of nine hepatic/splenic inflammatory pseudotumors expressing follicular dendritic cell markers and harboring Epstein-Barr virus, and none of seven inflammatory pseudotumors of the lymph node. When only those patients 40 years or younger were considered, the ALK positivity rate became 21.7% (five of 23). All five ALK+ cases occurred in young patients aged 0.5 to 37 years, who were alive and well at 3.5 to 17 years. The tumors exhibited a spectrum of histologic features typical of inflammatory pseudotumors/myofibroblastic tumors, but there was at least focal nuclear atypia. Immunostaining for ALK produced fibrillary or granular cytoplasmic staining in the neoplastic cells, sometimes with cell membrane accentuation. This study confirms that ALK is implicated in a proportion of inflammatory pseudotumors, and is generally associated with a favorable outcome. The results also support the heterogeneity of inflammatory pseudotumors, with the follicular dendritic cell/Epstein-Barr virus-positive cases and those occurring in lymph nodes representing different biologic entities. PMID- 11395554 TI - Lipomatous meningioma: a clinicopathologic study of 18 cases with special reference to the issue of metaplasia. AB - We report 18 cases of lipomatous meningioma occurring in patients aged 14 to 79, most being females (72%). Sixteen were supratentorial and 2 involved the spinal meninges. Follow-up ranged from 1 to 120 months. Fifteen patients were cured with surgery alone and 3 (17%) experienced a recurrence at 7, 8 and 24 months. Of these, one died with disease 4 years after resection of the primary lesion. Histologically, 12 tumors were meningothelial, 3 transitional, 2 showed myxoid stromal changes and 1 was microcystic. The 2 spinal tumors were atypical. The proportion of fatty cells ranged from 10 to 90%. These resembled mature adipocytes or less commonly lipoblasts. Xanthomatous meningothelial cells were also noted in 6 tumors (30%). Both conventional meningothelial as well as lipid laden cells exhibited epithelial membrane antigen immunoreactivity. In addition, occasional cells resembling mature adipocytes showed reactivity for S-100 protein. Ultrastructurally, lipidization of neoplastic cells varied from intracytoplasmic lipid droplets to a single massive globule. Moreover, lipid laden meningothelial cells featured interdigitating cell membranes and well formed desmosomes. Lipid droplets were not membrane-bound. In that metaplasia denotes differentiation of one mature cell type to another, lipid accumulation in meningiomas cannot be considered true metaplasia since their lipid-laden cells retain the immunophenotype and ultrastructural features of meningothelium. We suggest that this distinctive subset of meningiomas be termed "lipidized meningiomas" rather than being included in the metaplastic category. PMID- 11395555 TI - Primary monophasic synovial sarcoma of the pleura: five cases confirmed by the presence of SYT-SSX fusion transcript. AB - This study reports five cases of primary pleural monophasic synovial sarcomas and assesses the role of the SYT-SSX fusion transcript in the differential diagnosis. Patients had a mean age of 47 years with no gender predilection. Chest pain and pleural-based masses with effusions characterized the clinical presentations. Each patient underwent a complete surgical resection of the mass. The mean follow up was 9 months, available in four patients. They were all alive, with no evidence of disease. Histologically, neoplasms were composed of densely packed fusiform cells focally alternating with less cellular areas. No epithelial differentiation was identified at the hematoxylin and eosin level. Keratin and epithelial membrane antigen reactivity was focal and present in four and two tumors, respectively. There was no immunoreactivity for CD34. RT-PCR studies for the presence of a SYT-SSX1 or SYT-SSX2 fusion transcript were positive in every tumor. In comparison, 10 localized fibrous tumors were immunohistochemically negative for keratin and epithelial membrane antigen and positive for CD34. A SYT SSX fusion transcript was not identified in any of five localized fibrous tumors tested. Identification of the synovial sarcoma-specific chimeric transcript (SYT SSX1 or SYT-SSX2), in conjunction with immunoperoxidase studies, can be extremely helpful in identifying cases of pleural monophasic synovial sarcoma. PMID- 11395557 TI - Dermoid cyst of the testis: a study of five postpubertal cases, including a pilomatrixoma-like variant, with evidence supporting its separate classification from mature testicular teratoma. AB - It is controversial if the rare dermoid cyst of the testis should be classified as a variant of mature teratoma or separately. The spectrum of findings is also ill defined, as is the relationship of dermoid cyst to intratubular germ cell neoplasia of the unclassified type (IGCNU). This study therefore reports the findings in five testicular dermoid cysts that occurred in five patients, 17-42 years of age, who presented with testicular masses. Four lesions consisted of a keratin-filled cyst with a thickened wall, whereas one had islands of "shadow" squamous epithelial cells with superimposed calcification and ossification (pilomatrixoma-like variant). Hair was identified grossly in two cases. On microscopic examination, four tumors had hair follicles with sebaceous glands showing a typical, cutaneous-type orientation to an epidermal surface, although no hair shafts were present in two. In addition, the fibrous wall contained smooth muscle bundles (all tumors) and eccrine or apocrine sweat glands (4 tumors). In some cases there were also glands lined by ciliated epithelium (4 tumors, including the pilomatrixoma-like variant), intestinal mucosa (1 tumor), and bone (2 tumors). There was no cytologic atypia or apparent mitotic activity, and no case had IGCNU in the seminiferous tubules. All patients were clinical stage I and were treated by orchiectomy without adjuvant therapy. All were well on follow-up from 1.5 to 9.5 years later. This study supports that dermoid cyst may have noncutaneous teratomatous elements and that an important criterion for its diagnosis is the absence of IGCNU. It also supports that it should be categorized separately from mature testicular teratoma because of the malignant nature of the latter in postpubertal patients. These observations suggest that there are at least two pathways for testicular teratomas in postpubertal patients: the more common being through IGCNU by differentiation from an invasive malignant germ cell tumor and the less common one, taken by dermoid cyst, by direct transformation from a nonmalignant germ cell. PMID- 11395556 TI - Expression of melanocytic differentiation markers in malignant melanomas of the oral and sinonasal mucosa. AB - Malignant melanomas of the oral and sinonasal mucosa are rare tumors. Amelanotic variants can, on occasion, be difficult to recognize by routine light microscopy. Immunohistochemical studies may be needed for a final diagnosis. A number of new monoclonal antibodies to melanocytic differentiation antigens have been studied recently on primary cutaneous and metastatic melanoma. However, little is known about these antibodies for the diagnosis of mucosal melanomas. In this study the authors analyzed 79 oral and sinonasal mucosal melanomas of 65 patients. A total of 35 tumors originated from the oral mucosa (21 primary tumors, eight local recurrences, and six metastases) and 44 melanomas were from the sinonasal tract (27 primary tumors, nine local recurrences, and eight metastases). Immunohistochemical studies were performed on paraffin-embedded tissues, using the following antibodies: anti-S-100 protein, T311 (anti-tyrosinase), A103 (anti Mart-1/Melan-A), D5 (antimicrophthalmia-associated transcription factor), and HMB 45 (anti-gp100). Of 35 oral mucosal tumors, 34 (97%) were positive with anti-S 100 protein, 33 (94%) with T311, 30 (85%) with A103, 26 (74%) with D5, and 25 (71%) with HMB-45. All five desmoplastic melanomas of the oral mucosa were positive for S-100 protein, four for tyrosinase, and one each for HMB-45 and A103. No desmoplastic melanoma was positive with D5. All 44 sinonasal melanomas were positive for tyrosinase and Mart-1/Melan-A (100%). Forty-three (98%) were positive with HMB-45, 42 (95%) with anti-S-100 protein, and 40 (91%) with D5. These results reveal that T311 is the most sensitive marker for sinonasal melanomas and closely approaches the sensitivity of anti-S-100 protein for oral mucosal melanomas. For desmoplastic mucosal tumors, anti-S-100 protein remains the most sensitive marker. PMID- 11395558 TI - A clinicopathologic analysis of urothelial carcinomas diagnosed on prostate needle biopsy. AB - No data exist on urothelial carcinoma diagnosed on prostatic needle biopsy. We reviewed 21 cases (19 consultations) of urothelial carcinoma diagnosed on prostate needle biopsy from 1991 to 1998. In 13 of 21 (62%) cases, urothelial carcinoma showed in situ urothelial carcinoma involving prostatic ducts and acini (DCIS) only; 6 of 21 (29%) cases showed both DCIS and invasive carcinoma and 2 of 21 (9%) cases showed widespread stromal invasion without DCIS. In contrast to prostatic adenocarcinoma, cases exhibited greater nuclear pleomorphism, variably prominent nucleoli, increased mitoses, and necrosis. Immunostains for PSA and PSAP were negative in all 18 cases studied. CK7 was positive in 14 of 16 cases, CK20 was positive in 13 of 16 cases, and 34betaE12 was positive in 11 of 17 cases. A total of 7 of 17 (41%) men had no prior or subsequent history of urothelial carcinoma outside the prostate, 6 of 17 (35%) had concurrent urothelial cell carcinomas of the bladder (1 with extensive carcinoma in situ [CIS] at cystoprostatectomy), 2 of 17 (12%) had a prior urothelial cell carcinoma, and 2 of 17 (12%) developed urothelial cell carcinomas outside the prostate subsequent to the needle biopsy diagnosis. A total of 14 of 18 (78%) men had an elevated prostate specific antigen (PSA), abnormal digital rectal examination, or abnormal ultrasound suggestive of prostatic adenocarcinoma. Follow-up information was available in 17 cases. Six of nine (67%) patients with DCIS eventually died of disease (DOD) (2 with prior urothelial cell carcinoma, 1 with no prior or subsequent history, 3 without information), and 3 of 9 (33%) patients with DCIS were alive with residual disease (AWD). Of the patients with invasive carcinomas, 4 of 8 (50%) were DOD, 2 of 8 (25%) were AWD, and 2 of 8 (25%) were alive without evidence of disease. All men who are alive were treated aggressively with surgery and often adjuvant chemotherapy-radiation. Overall, 10 of 17 (59%) men were DOD with a mean survival after diagnosis of 23.2 months (2 72 months). The diagnosis of urothelial carcinoma on prostate needle biopsy is difficult because it is rare and clinically can mimic prostatic adenocarcinoma; often there is no history of urothelial carcinoma elsewhere. Although the prognosis is poor even with only apparent DCIS, histologic recognition is essential because the only opportunity for improved outcome is early and aggressive treatment. PMID- 11395559 TI - Nephrogenic adenoma of the prostatic urethra: a mimicker of prostate adenocarcinoma. AB - Nephrogenic adenoma, thought to be a benign metaplastic response of the urothelium to injury, can rarely affect the prostatic urethra. Extension of small tubules of nephrogenic adenoma into the underlying prostatic fibromuscular stroma can lead to the misdiagnosis of prostatic adenocarcinoma in transurethral resection specimens and prostate biopsies. We reviewed 26 cases of nephrogenic adenoma involving the prostatic urethra, seen at The Johns Hopkins Hospital from 1990 to 1998, to evaluate the histologic features, which may better define this lesion. Immunohistochemical results were evaluated for cases where the lesion was present on deeper sections. Histologic patterns included the following: tubules in 96% (25 of 26), structures resembling vessels in 73% (19 of 26), cords and individual cells in 46% (12 of 26), papillary configurations in 19% (5 of 26), and signet ring cell-like tubules in 12% (3 of 26). Other features of nephrogenic adenoma, such as thyroidization, were identified in 38% (10 of 26), and peritubular sheaths were seen in 65% (17 of 26) of cases. Nucleoli were prominent in 54% (14 of 26), and no case had mitoses. In the region of nephrogenic adenoma, urothelium was noted in 69% (18 of 26); in 61% (11 of 18) it showed cuboidal metaplasia and 28% (5 of 18) showed squamous metaplasia. Extension of nephrogenic adenoma into muscle was observed in 77% (20 of 26) of cases, 75% (15 of 20) of which had identifiable urothelium overlying the lesion. Blue-tinged mucinous secretions were observed in 32% (8 of 25) of cases. Inflammation was found in all but one case. Nephrogenic adenomas were diffusely positive for 34betaE12 in 11% (1 of 9) of cases, focally positive in 44% (4 of 9), and negative in 44% (4 of 9). In 100% (9 of 9), cytokeratin 7 stains were positive. Focal prostate specific antigen and PSAP positivity were seen in 36% (4 of 11) and 50% (5 of 10) of nephrogenic adenoma cases, respectively. In conclusion, nephrogenic adenoma of the prostatic urethra can mimic prostate cancer because of: 1) the presence of tubules, cords, and signet ring-like tubules; 2) prominent nucleoli; 3) muscle involvement; 4) blue-tinged mucinous secretions; 5) focal prostate specific antigen and PSAP positivity; and 6) negative staining in some cases for 34betaE12. Features useful in the diagnosis of nephrogenic adenoma include the following: 1) distinctive nephrogenic patterns, such as papillary and "vascular," 2) adjacent urothelium, 3) thyroidization, 4) peritubular sheaths, 5) associated inflammation, and 6) positivity for cytokeratin 7 and, in some cases, 34betaE12. PMID- 11395560 TI - The morphologic spectrum of hibernoma: a clinicopathologic study of 170 cases. AB - Hibernoma, an uncommon tumor of brown fat, has been described only in a few case reports and small series. The authors reviewed 170 cases of hibernoma and evaluated the morphologic features and the behavior of this tumor. The records from the Soft Tissue Registry of the Armed Forces Institute of Pathology from 1970 were searched for cases coded as "hibernoma." Clinical information and available slides from 170 hibernomas were reviewed. Immunohistochemical staining for S-100 and CD34 was performed on select cases. Follow-up information was obtained from the patients' medical records, the patients' physicians, and the patients themselves. Of 170 patients with hibernoma, 99 were men and 71 were women. The tumor occurred most commonly in adults, with a mean age of 38.0 years (age range, 2-75 years). Nine tumors occurred in pediatric patients. The most common anatomic locations included the thigh (n = 50), shoulder (n = 20), back (n = 17), neck (n = 16), chest (n = 11), arm (n = 11), and abdominal cavity/retroperitoneum (n = 10). The average duration of the tumor was 30.6 months. Tumor size ranged from 1 to 24 cm with an average dimension of 9.3 cm. All tumors were composed partly or principally of coarsely multivacuolated fat cells with small, central nuclei and no atypia. Four morphologic variants of hibernoma were identified: typical, myxoid, spindle cell, and lipoma-like. "Typical" hibernoma (n = 140) included eosinophilic cell, pale cell, and mixed cell types based on the tinctorial quality of the hibernoma cells. The myxoid variant (n = 14) contained a loose basophilic matrix. Spindle cell hibernoma (n = 4) had features of spindle cell lipoma and hibernoma; all occurred in the neck or scalp. The lipoma-like variant (n = 12) contained only scattered hibernoma cells. Immunohistochemically, 17 of 20 cases (85%) were positive for S-100 protein. Only one hibernoma of 20, a spindle cell variant, was positive for CD34, whereas other hibernoma variants were negative. Follow-up was obtained for 66 cases (39%) over a mean period of 7.7 years (range, 6 months-28 years). None of the patients with follow-up had a recurrence or metastasis, including eight with intramuscular tumors. No patient died of disease. Hibernoma is a tumor found most often in adults and most commonly in the thigh, with several morphologic variants. It is a benign tumor that does not recur with complete excision. Hibernomas should not be confused with atypical lipomas or well-differentiated liposarcoma. PMID- 11395561 TI - Thyroid transcription factor-1 distinguishes metastatic pulmonary from well differentiated neuroendocrine tumors of other sites. AB - Metastatic neuroendocrine neoplasms can have similar histologic appearances, and without an obvious primary, it may be difficult to determine the site of origin of the metastasis. Thyroid transcription factor-1 (TTF-1) is a nuclear protein expressed during the development of thyroid, lung, and forebrain. The clinical utility of TTF-1 to distinguishing between metastatic pulmonary and nonpulmonary well-differentiated neuroendocrine tumors (WDNET) has not been previously studied. One hundred fifty-eight primary and metastatic WDNET were evaluated for TTF-1 expression. The tumors included 20 pulmonary WDNET, including 17 typical and 3 atypical carcinoid tumors, 10 metastatic pulmonary WDNET, 26 intestinal WDNET, 24 metastatic intestinal WDNET, 3 thymic mediastinal WDNET, 30 thyroid tumors (10 medullary carcinomas, 5 follicular carcinomas, 5 follicular adenomas, 5 papillary carcinomas, and 5 anaplastic carcinomas), 10 parathyroid adenomas, 20 pituitary adenomas, 10 pancreatic WDNET, and 5 pheochromocytomas. TTF-1 expression was found in 19 of 20 (95%) pulmonary WDNET, 8 of 10 (80%) metastatic pulmonary WDNET, and in 0 of 50 (0%) intestinal WDNET. All thyroid tumors were diffusely positive for TTF-1, except for three anaplastic carcinomas. All parathyroid and pituitary adenomas, pancreatic and thymic WDNET, and pheochromocytomas were uniformly negative for TTF-1. These results indicate that TTF-1 is clinically useful in distinguishing metastatic pulmonary from metastatic WDNET of extrapulmonary origin. PMID- 11395562 TI - Transbronchial biopsy: nothing to fear. PMID- 11395563 TI - Solitary cylindroma (dermal analog tumor) of the breast: a previously undescribed neoplasm at this site. AB - The authors report a previously undescribed small, well-demarcated breast tumor similar to a dermal cylindroma in a 63-year-old woman. The tumor was an incidental finding in a lumpectomy specimen for infiltrating lobular carcinoma. The cylindroma was surrounded by normal-appearing breast parenchyma and had the typical "jigsaw" pattern of epithelial basaloid islands. The islands showed focal squamous and myoepithelial differentiation. A notable number of reactive dendritic Langerhans cells permeated the epithelial cell islands, a feature considered to be characteristic of dermal cylindroma. There was also ductal differentiation. Thick bands of hyaline periodic acid-Schiff (PAS) stain and collagen IV-positive basement membrane material bordered the cell islands, and PAS-collagen IV-positive hyaline globules were seen within the cell islands. There was no nuclear pleomorphism or mitotic figures. The cylindroma did not express gross cystic disease fluid protein 15, carcinoembryonic antigen, estrogen and progesterone receptors, or cytokeratin 20 (CK20). There was diffuse and strong immunoreactivity to CK AE1/AE3, and focal reactivity for CK7 and smooth muscle actin. Cylindroma of the breast should be distinguished from adenoid cystic carcinoma and basal cell carcinoma. Although clearly epithelial, the exact histogenesis and cell phenotype of this unusual dermal type cylindroma of the breast are unknown. PMID- 11395564 TI - Isolated regional vasculitis. PMID- 11395565 TI - Nuchal-type fibroma appearance in a desmoid fibromatosis. PMID- 11395566 TI - Pulmonary cytolytic thrombi (PCT). A previously unrecognized complication of bone marrow transplantation (BMT). PMID- 11395567 TI - Small cell carcinoma of the breast--additional immunohistochemical studies. PMID- 11395568 TI - The urokinase plasminogen activator receptor (uPAR) as a target for the diagnosis and therapy of cancer. AB - The identification and characterization of validated molecular targets for cancer drug and diagnostic development is rapidly changing the way that promising new anti-cancer compounds are developed and evaluated. A significant body of in vitro and in vivo data has established the urokinase plasminogen activator (uPA) system as a promising target for cancer drug development. The uPA system has been demonstrated to have pleiotropic activities in the development of tumors, and in tumor progression and angiogenesis. There are multiple ways to target this system, the most straightforward being the development of small molecule active site inhibitors of the serine protease, uPA. However, compounds of this type have not entered into clinical trials, and issues related to selectivity and specificity of this class of inhibitors have yet to be satisfactorily resolved. Recent evidence suggests that in addition to uPA, its specific cell surface receptor (uPAR) may also be a suitable target for the design and development of cancer therapeutic and diagnostic agents. uPAR is central to several pathways implicated in tumor progression and angiogenesis. The binding of the uPA zymogen (scuPA) to uPAR appears to be a pre-requisite for efficient cell-surface activation of scuPA to the active two-chain form (tcuPA) by plasmin, and simple ligand occupancy of uPAR by scuPA initiates various signaling pathways leading to alterations in cell motility and adhesion. One therapeutic rationale that is currently being investigated is the simple displacement of scuPA or tcuPA from suPAR, which may effectively inhibit both the proteolytic and signal-transducing cascades. In addition, other approaches to the modulation of the activity of this system that may also be useful include blocking the interaction of uPAR with integrins and extracellular matrix proteins as well as strategies to down regulate the expression of uPA and uPAR in target cells. This review will summarize these approaches, and also describe the targeting of uPAR for diagnosis and imaging. PMID- 11395569 TI - Quinone isomers of the WS-5995 antibiotics: synthetic antitumor agents that inhibit macromolecule synthesis, block nucleoside transport, induce DNA fragmentation, and decrease the growth and viability of L1210 leukemic cells more effectively than ellagic acid and genistein in vitro. AB - Antibiotic WS-5995A (code name J4) and two of its synthetic analogs, o-quinone J1 and model p-quinone J7, which show some structural similarity with both ellagic acid (EA) and genistein (GEN), were compared for their antileukemic activity in L1210 cells in vitro. Overall, J4 is more cytostatic and cytotoxic than J1 and J7, suggesting that methyl and methoxy substitutions, a p-quinone moiety, and a hydrogen bonding phenolic group may enhance the antitumor potential of these naphthoquinone lactones, which are all more potent than EA and GEN. For instance, the lead compound J4 inhibits tumor cell proliferation and viability at day 4 (IC(50): 0.24--0.65 microM) more effectively than EA (IC(50): 5--6 microM) and GEN (IC(50): 7 microM). Since J4 does not increase but rather decreases the mitotic index of L1210 cells at 24 h, it is not an antitubulin drug but might arrest early stages of cell cycle progression like EA and GEN. A 1.5- to 3-h pretreatment with J4 is sufficient to inhibit the rates of DNA, RNA and protein syntheses (IC(50): 2.0--2.5 microM) determined over 30- to 60-min periods of pulse-labeling in L1210 cells in vitro, whereas EA (IC(50): 20-130 microM) and GEN (IC(50): 40--115 microM) are less effective against macromolecule synthesis. In contrast to 156 microM EA, which is inactive, a 15-min pretreatment with 10- 25 microM J4 has the advantage of also inhibiting the cellular transport of both purine and pyrimidine nucleosides over a 30 s period in vitro, an effect which can be mimicked by 156 microM GEN. Hence, the WS-5995 analogs and GEN may prevent the incorporation of [(3)H]adenosine and [(3)H]thymidine into DNA because they rapidly block the uptake of these nucleosides by the tumor cells. After 24 h, the concentration-dependent induction of DNA cleavage by J4 peaks at 10 microM and declines at 25 microM, whereas EA and GEN are ineffective at 10 microM but maximally stimulate DNA cleavage at 62.5 microM. Like EA and GEN, the mechanism by which J4 induces DNA fragmentation is inhibited by actinomycin D, cycloheximide, benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone, N-tosyl-L phenylalanine chloromethyl ketone and ZnSO(4), suggesting that J4 triggers apoptosis by caspase and endonuclease activation. Because they are more potent than EA and GEN, and affect both nucleoside transport and DNA cleavage, the WS 5995 antitumor antibiotics might be valuable in polychemotherapy to potentiate the action of antimetabolites and sensitize multidrug-resistant tumor cells. PMID- 11395570 TI - Active transepithelial transport of irinotecan (CPT-11) and its metabolites by human intestinal Caco-2 cells. AB - Irinotecan (CPT-11) is a camptothecin analog with low (about 10--20%) and variable oral bioavailability in animal models. Here, Caco-2 cells were used to evaluate the transepithelial transport of CPT-11 and its metabolites. Caco-2 cells demonstrated significant expression of P-glycoprotein (P-gp), multidrug resistance-associated protein and canalicular multispecific organic anion transporter. Both the lactone and carboxylate forms of CPT-11 and SN-38 were actively transported across the cell monolayers, mainly by the apical-localized P gp pump. Cellular permeability of CPT-11 at a concentration of 17 microM converted from active to passive-diffusional transport between the 2 and 6 h exposure time points. Antiproliferative effects of CPT-11 were related to permeability of the lactone form, whereas for SN-38 efficacy was dependent on lactone accumulation. Exposure of CPT-11 with cyclosporin A significantly enhanced its efficacy, whereas this was not observed with verapamil and R101933. In contrast, SN-38 efficacy decreased in the presence of P-gp inhibitors due to active transport toward the basolateral side, thereby reducing drug accumulation. Hence, multiple-active transport systems could be demonstrated to be responsible for not only accumulation profiles but also cytotoxic efficacy of CPT-11 and SN 38 in the intestinal Caco-2 cells. It is suggested that CPT-11 might act in a time-dependent manner and that SN-38-mediated cytotoxicity relates to (dose dependent) lactone kinetics. The results detailed in this report could contribute toward the development of a clinically useful oral formulation of CPT-11 with improved absorption characteristics and suggest that cyclosporin A is a suitable agent for further research of this concept. PMID- 11395571 TI - Antitumor bicyclic hexapeptide RA-VII modulates cyclin D1 protein level. AB - A bicyclic hexapeptide, RA-VII or O-methyl deoxybouvardin, isolated from Rubia cordifolia, is known to inhibit protein biosynthesis in vitro and in vivo. We here demonstrate that the treatment of human colon cancer DLD-1 cells with RA-VII induces cell growth inhibition associated with a partial G1 arrest and a rapid decrease (below 2 h) in the level of cyclin D1 protein. Since cycloheximide, another protein synthesis inhibitor, neither decreased the amount of cyclin D1 in the cells nor arrested cells in G1 phase, it is unlikely that this RA-VII-induced reduction of cyclin D1 was fully dependent on its direct inhibitory effect of protein synthesis. Northern blot analysis revealed that RA-VII did not affect the level of cyclin D1 mRNA. Meanwhile, pre-treatment of cells with lactacystin, a proteasome inhibitor, abolished the RA-VII-induced decrease in cyclin D1. Moreover, RA-VII still decreased cyclin D1 protein in the presence of cycloheximide. These results indicate that the RA-VII-induced cyclin D1 decrease depends on cyclin D1 degradation via the ubiquitin-proteasome pathway and does not require additional protein synthesis. RA-VII might actively proceed the degradation process of cyclin D1 via the ubiquitin-proteasome pathway in DLD-1 cells. PMID- 11395572 TI - Altered DNA-cleavage activity of topoisomerase II from WEHI-3B leukemia cells with specific resistance to ciprofloxacin. AB - In order to investigate the mechanisms of drug resistance arising in tumor cells, we investigated the capacity of fluoroquinolones to inhibit the in vitro growth of WEHI-3B monomyelocytic leukemia cells and then we established a variant of this line (currently maintained in the absence of drug). The line, named WEHI 3B/CPX, expresses a specific resistance to ciprofloxacin (CPX; resistance index=17.3+/-2.2), and does not show cross-resistance with other fluoroquinolones, camptothecin and topoisomerase II inhibitors such as doxorubicin, etoposide and teniposide. Although a little decrease in intracellular accumulation of CPX is observed in WEHI-3B/CPX cells, these cells do not express MDR or LRP markers, and the resistance is not circumvented by verapamil. Purified nuclear extracts from WEHI-3B and WEHI-3B/CPX cells were tested for topoisomerase I catalytic activity and checking in vitro topoisomerase I sensitivity to CPX and camptothecin inhibition, but no difference was observed. As the treatment with CPX showed that the resistant cell line suffers a significantly lower number of breaks in the DNA molecule we also addressed our investigations to the topoisomerase II-dependent DNA cleavage that, in the resistant clone, was found dramatically less susceptible to be enhanced by CPX both in pre-strand and post-strand DNA passage conditions. WEHI-3B/CPX cells do not express any character of multidrug resistance and represent a rare case of specific drug resistance to CPX. The specific resistance to CPX observed in these cells is related to a functional decrease of topoisomerase II cleavage activity. It could be consequent to a decreased binding affinity of CPX for the topoisomerase II--DNA complex or to a decreased affinity or specificity of topoisomerase II for its DNA cleavage sites. PMID- 11395573 TI - (S)-(-)-Bromofosfamide (CBM-11): synthesis and antitumor activity and toxicity in mice. AB - (S)-(-)-Bromofosfamide (CBM-11), an enantiomerically pure bromo analog of ifosfamide, was found to be potent against several model tumors in mice. Therapeutic indices of CBM-11 were more favorable as compared to those received for ifosfamide. PMID- 11395574 TI - The bisphosphonate pamidronate is a potent inhibitor of human osteosarcoma cell growth in vitro. AB - Bisphosphonates (BPs), such as pamidronate and clodronate, are an important class of drugs for the treatment of bone diseases. It is widely recognized that they inhibit bone resorption by suppressing the action of osteoclasts through antagonizing the mevalonate pathway, thereby reducing osteolytic bone metastases derived from different cancers, i.e. breast carcinoma and multiple myeloma. In contrast, the effects of BPs on primary bone tumors is an issue still to be resolved. Therefore, a systematic approach was set up to test the hypothesis that BPs could act directly on osteosarcoma cells. The effects of pamidronate and clodronate on seven osteosarcoma cell lines (HOS, MG-63, OST, SaOS-2, SJSA-1, U(2)OS and ZK-58) were studied. Pamidronate inhibited cell growth in a time- and dose-dependent manner, and decreased proliferation for up to 73% at 50 microM after 72 h, whereas its monophosphonate analog 3-aminopropyl phosphonate did not reduce cell viability at concentrations up to 2 mM. Clodronate showed less inhibitory effects (maximally 38% reduction at 1 mM after 72 h). Importantly, cell growth of fibroblasts was only very weakly affected by treatment with pamidronate. These results suggest that pamidronate may be a useful agent for the treatment of patients with osteosarcoma. PMID- 11395575 TI - Treatment of drug-resistant human neuroblastoma cells with cyclodextrin inclusion complexes of aphidicolin. AB - Treatment failure in most neuroblastoma (NB) patients is related to primary and/or acquired resistance to conventional chemotherapeutic agents. Aphidicolin (APH), a tetracyclic diterpene, exhibits specific cytotoxic action against NB cells. The purpose of this study was to compare antitumoral efficacy of APH in parental NB cell lines and cell subclones that exhibit drug resistance to vincristine (VCR), doxorubicin (DOX) and cisplatin. Due to poor solubility of APH in water, gamma-cyclodextrin (gamma-CD) inclusion complexes of APH were used for systemic treatment of xenotransplanted parental and VCR-resistant UKF-NB-3 tumours. APH and its gamma-CD inclusion complexes inhibited growth of parental and drug-resistant NB cells at equimolar doses in vitro. Growth of VCR-sensitive and -resistant NB tumors was inhibited at equal doses in a dose-dependent fashion in vivo. These results indicate that the specific cytotoxic activity of APH against NB cells in vitro and in vivo is independent of cellular mechanisms facilitating drug resistance to conventional chemotherapeutic drugs. Hence, taking into account our previous findings that APH acts synergistically with VCR and DOX, APH might be an additive tool for the therapy of NB and is suitable for evaluation in clinical studies of NB treatment protocols. PMID- 11395576 TI - Cytotoxicity of digitoxin and related cardiac glycosides in human tumor cells. AB - The saponin digitonin, the aglycone digitoxigenin and five cardiac glycosides were evaluated for cytotoxicity using primary cultures of tumor cells from patients and a human cell line panel (representing different cytotoxic drug resistance patterns). Of these seven compounds, proscillaridin A was the most potent (IC(50): 6.4--76 nM), followed by digitoxin, and then ouabain, digoxin, lanatoside C, digitoxigenin and digitonin. Correlation analysis of the log IC(50) values for the cell lines in the panel showed that compound cytotoxicity was only slightly influenced by resistance mechanisms that involved P-glycoprotein, topoisomerase II, multidrug resistance-associated protein and glutathione mediated drug resistance. Digitoxin and digoxin expressed selective toxicity against solid tumor cells from patients, while proscillaridin A expressed no selective toxicity against either solid or hematological tumor cells. The results revealed marked differences in cytotoxicity between the cardiac glycosides, both in potency and selectivity, and modes of action for cytotoxicity that differ from that of commonly used anticancer drugs. PMID- 11395577 TI - Pharmacokinetics of paclitaxel and cisplatin in a hemodialysis patient with recurrent ovarian cancer. AB - This is the first report that the combination of paclitaxel and cisplatin is feasible in a patient with recurrent ovarian cancer undergoing hemodialysis. Paclitaxel at a dose of 150 mg/m(2) was administered as a 3-h continuous i.v. infusion. Thirty minutes after paclitaxel administration, cisplatin was administered at a dose of 30 mg/m(2) for 30 min. Hemodialysis was started 30 min after completion of the cisplatin infusion and performed for 5 h. The maximum plasma concentrations of paclitaxel, total platinum and free platinum were 3.26, 2.44 and 1.84 microg/ml, respectively. The AUC of paclitaxel and free platinum were 15.3 and 1.76 microg x h/ml, respectively. The pelvic tumor size was reduced by 42% on MRI after the second course of this therapy. Grade IV neutropenia and grade III thrombopenia were observed. We conclude that paclitaxel and cisplatin combination chemotherapy is efficacious and feasible for an ovarian cancer patient under hemodialysis. PMID- 11395583 TI - Impact of a rotating empiric antibiotic schedule on infectious mortality in an intensive care unit. AB - OBJECTIVE: The development of antibiotic-resistant bacteria is associated with significant morbidity and mortality in critically ill patients. We postulated that quarterly rotation of empirical antibiotics could decrease infectious complications from resistant organisms in an intensive care unit (ICU). DESIGN: Prospective cohort study. SETTING: An ICU at a university medical center. SUBJECTS: All patients admitted to the general, transplant, or trauma surgery services who developed pneumonia, peritonitis, or sepsis of unknown origin. INTERVENTIONS: A 2-yr study consisting of 1 yr of nonprotocol-driven antibiotic use and 1 yr of rotating empirical antibiotic assignment. MEASUREMENTS AND MAIN RESULTS: Over 100 variables were recorded for each infectious episode, including patient characteristics (e.g., Acute Physiology and Chronic Health Evaluation [APACHE] II score, age, comorbidities), infection characteristics (e.g., site, organism), treatment characteristics (e.g., antibiotic, treatment duration) and outcome measures (e.g., mortality, length of stay, antibiotic cost). Of 1456 consecutive admissions to the ICU, 540 episodes of infection were treated. No differences were noted in age, APACHE II score, race, overall antibiotic utilization or duration of therapy between the 2 yrs of study. Outcome analysis revealed significant reductions in the incidence of antibiotic-resistant Gram positive coccal infections (7.8 infections/100 admissions vs. 14.6 infections/100 admissions, p <.0001), antibiotic-resistant Gram-negative bacillary infections (2.5 infections/100 admissions vs. 7.7 infections/100 admissions, p <.0001), and mortality associated with infection (2.9 deaths/100 admissions vs. 9.6 deaths/100 admissions, p <.0001) during rotation. Logistic regression identified age (odds ratio [OR], 1.03; 95% confidence interval [CI], 1.01-1.06), APACHE II score (OR, 1.06; 95% CI, 1.01-1.13), solid organ transplantation (OR, 9.50; 95% CI, 2.01 52.21), and malignancy (OR, 10.16; 95% CI, 4.11-26.96) as independent predictors of mortality. Antibiotic rotation was an independent predictor of survival (OR 6.27, 95% CI 2.78-14.16). CONCLUSION: Rotation of empirical antibiotic therapy seems to be a promising method to reduce infectious mortality in an ICU. PMID- 11395584 TI - Experience with a clinical guideline for the treatment of ventilator-associated pneumonia. AB - OBJECTIVE: To evaluate a clinical guideline for the treatment of ventilator associated pneumonia. DESIGN: Prospective before-and-after study design. SETTING: A medical intensive care unit from a university-affiliated, urban teaching hospital. PATIENTS: Between April 1999 and January 2000, 102 patients were prospectively evaluated. INTERVENTIONS: Prospective patient surveillance, data collection, and implementation of an antimicrobial guideline for the treatment of ventilator-associated pneumonia. MEASUREMENTS AND MAIN RESULTS: The main outcome evaluated was the initial administration of adequate antimicrobial treatment as determined by respiratory tract cultures. Secondary outcomes evaluated included the duration of antimicrobial treatment for ventilator-associated pneumonia, hospital mortality, intensive care unit and hospital lengths of stay, and the occurrence of a second episode of ventilator-associated pneumonia. Fifty consecutive patients with ventilator-associated pneumonia were evaluated in the before period and 52 consecutive patients with ventilator-associated pneumonia were evaluated in the after period. Severity of illness using Acute Physiology and Chronic Health Evaluation II (25.8 +/- 5.7 vs. 25.4 +/- 8.1, p =.798) and the clinical pulmonary infection scores (6.6 +/- 1.0 vs. 6.9 +/- 1.2, p =.105) were similar for patients during the two treatment periods. The initial administration of adequate antimicrobial treatment was statistically greater during the after period compared with the before period (94.2% vs. 48.0%, p <.001). The duration of antimicrobial treatment was statistically shorter during the after period compared with the before period (8.6 +/- 5.1 days vs. 14.8 +/- 8.1 days, p <.001). A second episode of ventilator-associated pneumonia occurred statistically less often among patients in the after period (7.7% vs. 24.0%, p =.030). CONCLUSIONS: The application of a clinical guideline for the treatment of ventilator-associated pneumonia can increase the initial administration of adequate antimicrobial treatment and decrease the overall duration of antibiotic treatment. These findings suggest that similar types of guidelines employing local microbiological data can be used to improve overall antibiotic utilization for the treatment of ventilator-associated pneumonia. PMID- 11395585 TI - Secondary insults in severe head injury--do multiply injured patients do worse? AB - OBJECTIVES: To study the occurrence of secondary insults and the influence of extracranial injuries on cerebral oxygenation and outcome in patients with closed severe head injury (Glasgow Coma Scale score < or =8). DESIGN: Two-year prospective, clinical study. SETTING: Two intensive care units in a level III trauma center. PATIENTS: We studied 119 patients. Eighty patients had severe head injury and were divided into two categories: "isolated" severe head injury patients (n = 36, Injury Severity Score <30), and severe head injury patients with associated extracranial injuries (n = 44, Injury Severity Score >29). Thirty nine patients with extracranial injuries and no head injury served as the control group. INTERVENTIONS: After patients were admitted to the intensive care unit, we began continuous multimodal cerebral monitoring of intracranial pressure, mean arterial blood pressure, cerebral perfusion pressure, end-tidal Co2, brain tissue Po2 (Licox), jugular bulb oxyhemoglobin saturation in severe head injury patients, and mean arterial blood pressure in the control group. Targets of management included intracranial pressure <20 mm Hg, cerebral perfusion pressure >60 mm Hg, Paco2 > 30 mm Hg, control of cerebral oxygenation, and delayed surgery for non-life-threatening extracranial lesions. MEASUREMENTS AND MAIN RESULTS: Data were analyzed for critical thresholds. The occurrence of secondary insults (intracranial pressure >20 mm Hg, mean arterial blood pressure <70 mm Hg, cerebral perfusion pressure <60 mm Hg, end-tidal Co2 <30 torr, brain tissue Po2 <10 torr, jugular bulb oxyhemoglobin saturation <50%) was comparable in patients with isolated severe head injury and those with severe head injury with associated extracranial lesions (Abbreviated Injury Scale score < or =5). The duration of intracranial hypertension and arterial hypotension significantly correlated with an unfavorable outcome, independent of the Injury Severity Score. In patients with severe head injury, 1-yr outcome was 29% dead or vegetative, 17% severely disabled, and 54% moderate or good outcome. This was similar to patients with severe head injury and extracranial injuries (31% dead or vegetative, 14% severely disabled, and 56% moderate or good outcome) and was independent of the Injury Severity Score. Patients with no head injury had less secondary insults (mean arterial blood pressure <70 mm Hg, p <.01) and a better outcome compared with both severe head injury groups (p <.044). CONCLUSIONS: In patients with severe head injury who have targeted management including intracranial pressure- and cerebral perfusion pressure-guided therapy and delayed surgery for extracranial lesions, the occurrence of secondary insults in the intensive care unit and long-term neurological outcome were comparable and independent of the presence of extracranial lesions (Abbreviated Injury Severity level < or =5). A severe head injury is still a major contributor predicting an unfavorable outcome in multiply injured patients. PMID- 11395586 TI - Angiographic correlates of a positive troponin T test in patients with unstable angina. AB - OBJECTIVE: To study the angiographic correlates of cardiac troponin T (cTnT) positive and -negative patients with unstable angina pectoris. BACKGROUND: A positive cTnT test identifies a high-risk subgroup of unstable angina pectoris patients. Only the high-risk cTnT-positive patients seem to benefit from a more aggressive antithrombotic treatment regimen. The underlying coronary pathology in cTnT-positive and -negative patients that explains the predictive power of cTnT on prognosis and response to antithrombotic therapy is largely unknown. METHODS: A total of 197 subsequently admitted patients with unstable angina pectoris underwent cTnT testing by a rapid bedside assay and early qualitative and quantitative angiography. Long-term follow-up was 12 months. RESULTS: Patients with cTnT-positive tests revealed more critical stenoses of culprit lesions (p =.041), more severe reductions of thrombolysis in myocardial infarction flow grades (p <.037), a higher prevalence of intracoronary thrombus (p =.079), and a poorer left ventricular function (p =.047). The odds ratio of cTnT was 5.8 (p <.0001) for presence of thrombus, reduced thrombolysis in myocardial infarction flow, and/or critical stenosis (>90%), and was 3.1 (p =.005) for presence of three-vessel disease, left main disease, and/or reduced left ventricular ejection fraction. Coronary bypass grafting was more frequently performed in the cTnT positive group. However, event-free survival was not different in our cohort characterized by a high rate of percutaneous coronary interventions. CONCLUSIONS: A positive cTnT test in patients with unstable angina pectoris indicates presence of more severe coronary artery disease and poorer left ventricular function. This finding could explain the differences in short- and long-term outcome and treatment responses to antithrombotic regimens. PMID- 11395588 TI - Transthoracic monitoring of the impedance of the right lung in patients with cardiogenic pulmonary edema. AB - OBJECTIVE: To evaluate the suitability of the new electrical impedance monitor RS 205 for monitoring of cardiogenic pulmonary edema (CPE). DESIGN: Prospective, controlled study. SETTING: A department of internal medicine in a 1,200-bed university-affiliated, teaching hospital. PATIENTS: Sixty patients, aged 52-80 yrs, 30 without CPE (controls) and 30 with or at high risk for CPE. INTERVENTIONS: Internal thoracic impedance (ITI) was monitored by the RS-205. The RS-205 is approximately three times more sensitive than the Kubicek monitor, and it eliminates the effect of the drift of skin-to-electrode impedance. This is achieved by eliminating skin electrode impedance by a special algorithm, thus allowing measurement of ITI rather than total transthoracic impedance. Measuring ITI, the main component of which is lung impedance, is a noninvasive and safe method. CPE was diagnosed in accordance with well-accepted clinical and roentgenological criteria. MEASUREMENTS AND MAIN RESULTS: The controls' initial ITI was 68.3 +/- 12.38 ohms. During 6 hrs of monitoring, the ITI attained a minimum average value of -1.3 +/- 2.08% and a maximum average value of 4.6 +/- 3.56% relative to baseline. In all patients entering CPE, ITI decreased by 14.4 +/- 5.42% on the average (p <.001) 1 hr before the appearance of clinical symptoms. In patients with evolving CPE, ITI decreased significantly compared with controls (22.25 +/- 9.82%, p <.001). In patients at the peak of pulmonary edema, ITI was 2.1 times lower than in the control group (33.1 +/- 10.90 ohms, p <.001). In the last hour before the resolution of CPE, ITI increased in all patients by 17.7 +/- 19.74% compared with the peak of disease (p <.05). After the resolution of pulmonary edema, ITI increased in all patients by 44.14 +/- 26.90% compared with the peak of disease (p <.001). Importantly, the trend in ITI in all patients changed in accordance with the dynamics of CPE. A mixed general linear model shows that ITI values correlated well with the degree of crepitation, a direct characteristics of CPE. CONCLUSIONS: The RS-205 is suitable for monitoring patients at high risk of CPE development. It enables detection of CPE and the monitoring of patients at all stages of CPE. PMID- 11395590 TI - Association of pulmonary artery catheter use with in-hospital mortality. AB - OBJECTIVE: To determine the association of pulmonary artery catheter (PAC) use with in-hospital mortality. DESIGN: Prospective, observational study. SETTING: The medical intensive care units (MICU) of two teaching hospitals. METHODS: The study included 751 adults who were admitted to the MICU, excluding those who stayed for <24 hrs. Demographics and the worst Acute Physiology and Chronic Health Evaluation (APACHE) II score within the first 24 hrs of MICU admission were obtained. Daily logistic organ dysfunction system (LODS) scores were calculated. The associations of in-hospital mortality with the admission source, admission disease category, APACHE II scores, the worst LODS scores, mechanical ventilation, and PAC use were determined using chi-square, Mann-Whitney U, and multiple logistic regression analysis tests. p Values < 0.05 were considered significant. RESULTS: Mean patient age was 52.6 +/- 17.1 yrs; 425 (57%) were male; 464 (62%) were African-American, 275 (37%) Caucasian, 6 (1%) Asian, and 6 (1%) Hispanic. PAC was used in 119/751 (16%). The median APACHE II and worst LODS scores were 19 and 4, respectively. The in-hospital mortality rate was 159/751 (21%). The median APACHE II score for survivors was 17.5, compared with 28.0 for nonsurvivors (p <.0001). The worst median LODS score was 4 for survivors, compared with 11 for nonsurvivors (p <.0001). Sixty-four (54%) of the 119 patients with PAC died, compared with 95 (15%) of the 632 without PAC (p <.0001). Multiple logistic regression analysis showed that higher APACHE II-predicted mortality rate (p =.0088) and worst daily LODS score (p <.0001) were associated with increased mortality. The admission source, admission disease category, PAC use, and mechanical ventilation were not associated with in-hospital mortality. CONCLUSIONS: This study could not detect an association between PAC use and mortality. The APACHE II-predicted mortality rate and the development of multiple organ dysfunction were the main determinants of poor outcome in critically ill patients admitted to MICU. PMID- 11395591 TI - Amiodarone versus diltiazem for rate control in critically ill patients with atrial tachyarrhythmias. AB - OBJECTIVE: To compare the rate-lowering effect of diltiazem and two amiodarone regimens in critically ill patients with recent-onset atrial tachyarrhythmias. DESIGN: Prospective, randomized, controlled study. SETTING: Medical cardiologic intensive care unit in a university hospital. PATIENTS: Sixty critically ill patients (Acute Physiology and Chronic Health Evaluation [APACHE] III score 70 +/ 30, age 67 +/- 10 yrs). INTERVENTIONS: Patients with atrial fibrillation (n = 57), atrial flutter (n = 2), or atrial tachycardia (n = 1, and a heart rate consistently >120 beats/min over 30 mins were randomly assigned to one of three intravenous treatment regimens. Group 1 received diltiazem in a 25-mg bolus followed by a continuous infusion of 20 mg/hr for 24 hrs, group 2 received amiodarone in a 300-mg bolus, and group 3 received amiodarone in a 300-mg bolus followed by 45 mg/hr for 24 hrs. MEASUREMENTS AND MAIN RESULTS: The primary study end point was a >30% rate reduction within 4 hrs. The secondary study end point was a heart rate <120 beats/min (a patient was considered to have uncontrolled tachycardia if heart rate was >120 beats/min 4 hrs after study drug). The primary study end point was achieved in 14/20 (70%), 11/20 (55%), and 15/20 (75%) of patients in groups 1, 2, and 3, respectively (chi2 = 1.95, p =.38). Uncontrolled tachycardia was more frequently observed in group 2 (0/20, 9/29 [55%], and 1/20 [5%] of patients in groups 1, 2, and 3, respectively; chi2 = 17, p =.00016). In patients achieving tachycardia control, diltiazem showed a significantly better rate reduction (p =.0001 group 1 vs. group 3, p =.0001 over time; p =.0001 group 1 vs. group 2, p =.001 over time) when compared with the amiodarone groups. Premature drug discontinuation due to hypotension was required significantly more often in group 1 (6/20 [30%], 0/20, and 1/20 [5%] for groups 1, 2, and 3, respectively; chi2 = 10, p =.01). CONCLUSION: Sufficient rate control can be achieved in critically ill patients with atrial tachyarrhythmias using either diltiazem or amiodarone. Although diltiazem allowed for significantly better 24 hr heart rate control, this effect was offset by a significantly higher incidence of hypotension requiring discontinuation of the drug. Amiodarone may be an alternative in patients with severe hemodynamic compromise. PMID- 11395592 TI - Positive end-expiratory pressure titration in acute respiratory distress syndrome patients: impact on right ventricular outflow impedance evaluated by pulmonary artery Doppler flow velocity measurements. AB - OBJECTIVE: Positive end-expiratory pressure (PEEP) titration in acute respiratory distress syndrome patients remains debatable. We used two mechanical approaches, calculation of the compliance of the respiratory system and determination of the lower inflexion point of the pressure-volume curve of the respiratory system, to identify specific PEEPs (PEEPS and PEEPA) whose impact on right ventricular (RV) outflow was compared with Doppler analysis of pulmonary artery flow velocity. DESIGN: Prospective, open, clinical study. SETTING: Medical intensive care unit of a university hospital. PATIENTS: Sixteen consecutive ventilator-dependent acute respiratory distress syndrome patients. INTERVENTIONS: Two PEEPs were determined: PEEPS was the highest PEEP associated with the highest value of respiratory compliance, and PEEPA was the coordinate of the lower inflexion point of the inspiratory pressure-volume curve on the pressure axis plus 2 cm H2O. MEASUREMENTS AND MAIN RESULTS: We observed a large difference between the two PEEPs, with PEEPA (13 + 4 cm H2O) > PEEPS (6 + 3 cm H2O). Changes in RV outflow impedance produced by tidal ventilation with zero end-expiratory pressure (ZEEP) and after application of these two PEEPs were assessed by Doppler study of pulmonary artery flow velocity obtained by a transesophageal approach, with particular reference to the end-expiratory and end-inspiratory pulmonary artery velocity-time integral, as reflecting RV stroke output, and mean acceleration as reflecting RV outflow impedance during an unchanged flow period. A significant inspiratory reduction in pulmonary artery velocity-time integral (from 11.8 + 0.3 to 10.0 + 0.3 cm) and mean acceleration (from 11.9 + 0.9 to 8.0 + 0.9 m/sec2) was observed with ZEEP, showing a reduction in RV stroke index (from 29.0 + 0.9 to 26.0 + 0.6 cm3/m2) by a sudden increase in outflow impedance during tidal ventilation. Application of PEEPA, which improved Pao2 (102 + 40 vs. 65 + 18 torr with ZEEP), worsened the inspiratory drop in RV stroke index (21.6 + 0.8 cm3/m2), resulting in a significant reduction in cardiac index compared with ZEEP (from 3.0 + 1.0 to 2.7 + 1.1). Application of PEEPS, which also significantly improved Pao2 (81 + 21 torr), was associated with a lesser impact on RV outflow impedance (inspiratory mean acceleration: 9.5 + 1 m/sec2) and cardiac index (3.2 + 1.0) than PEEPA. CONCLUSION: RV outflow impedance evaluated by the Doppler technique appeared sensitive to PEEP titration. Application of PEEPA worsened RV systolic function impairment produced by tidal ventilation. Conversely, application of PEEPS reduced RV systolic function impairment, suggesting an association with a lower pulmonary vascular resistance. PMID- 11395594 TI - Indocyanine green elimination rate detects hepatocellular dysfunction early in septic shock and correlates with survival. AB - OBJECTIVE: To determine whether indocyanine green clearance is an early indicator of hepatocellular injury in septic shock and to assess its predictive value. DESIGN: Observational study with prospective data collection. SETTING: Traumatology and critical care unit in a city hospital, staffed by traumatology and intensive care clinicians. PATIENTS: Twelve patients in septic shock who survived at least 2 months (group S) and nine patients who died within 2 wks (group N). INTERVENTIONS: Routine resuscitation from septic shock (surgery, fluid loading, and administration of catecholamines and antibiotic drugs). MEASUREMENTS AND MAIN RESULTS: Pulmonary artery occlusion pressure, cardiac index, oxygen delivery index, oxygen consumption index, and the indocyanine green elimination rate constant (KICG; or the slope of the loge [indocyanine green concentration] vs. time curve) 3-9 mins after injection were measured within 12 hrs of the onset of hypotension, then at 24 hrs, and every 24 hrs thereafter. Alanine aminotransferase and total bilirubin were measured on day 0 and day 1. Volume of fluid administered and duration of shock were the same in survivors and nonsurvivors. The oxygen consumption index was higher in survivors at 12 hrs, but no intergroup difference in pulmonary artery occlusion pressure, cardiac index, or oxygen delivery index was significant at any time point. KICG in nonsurvivors was lower than in survivors both initially and after 24 hrs, and it was subnormal in all patients except one survivor (p <.05). The KICG increased between 24 and 120 hrs in 11 survivors but progressively decreased and remained below 0.05 in seven nonsurvivors. The remaining two nonsurvivors died within 24 hrs of the initial measurement of KICG, which was >0.05. Alanine aminotransferase and total bilirubin were less sensitive measures of hepatic dysfunction in the first 24 hrs than the KICG. CONCLUSIONS: The KICG can identify reversible liver injury in septic shock, suggesting good prognosis. Either failure to increase the KICG within 120 hrs or an extremely low KICG is a poor prognostic sign. PMID- 11395595 TI - An enhanced fibrinolysis prevents the development of multiple organ failure in disseminated intravascular coagulation in spite of much activation of blood coagulation. AB - OBJECTIVES: To investigate the relationship between fibrinolytic enhancement and the development of multiple organ failure (MOF) in disseminated intravascular coagulation (DIC). To detect the useful prognostic index for outcome in DIC. DESIGN: Case-control study. SETTING: A department of internal medicine in a university hospital, a clinical division for diagnosis and treatment, mainly of respiratory diseases, hematologic diseases, DIC, and other diseases requiring critical care medicine. PATIENTS: A total of 69 DIC patients, 31 with MOF. INTERVENTIONS: None. MEASUREMENT AND MAIN RESULTS: The DIC patients with MOF had more elevated levels of tissue plasminogen activator antigen (t-PA) and plasminogen activator inhibitor antigen (PAI), and more depressed levels of plasmin-alpha2 plasmin inhibitor complex (PIC) and fibrin/fibrinogen degradation products than those without MOF, although no significant difference in thrombin antithrombin complex (TAT) levels was observed. A fibrinolytic enhancement (shown by PIC) was parallel to an activation of blood coagulation (shown by TAT) in DIC patients without MOF, although no such fibrinolytic enhancement was provoked even by much activation of blood coagulation in DIC patients with MOF. Whereas all the patients without MOF were restored from DIC, 14 of 31 patients with MOF were unable to be restored from DIC and died. A significant increase in plasma levels of t-PA and PAI under the condition of sustained hemostatic activation was observed in the patients who died. CONCLUSION: Enhanced fibrinolysis was considered to be the important defense mechanism in preventing the development of MOF in DIC. The increases in plasma levels of t-PA and PAI were poor prognostic markers in DIC. Further careful study may be useful to clarify whether the fibrinolytic therapy is beneficial in clinical DIC patients with MOF. PMID- 11395597 TI - Inhaled nitric oxide down-regulates intrapulmonary nitric oxide production in lipopolysaccharide-induced acute lung injury. AB - OBJECTIVE: To examine whether inhaled nitric oxide (NO) affected the intrapulmonary production of NO, reactive oxygen species, and nuclear factor kappaB in a lipopolysaccharide (LPS)-induced model of acute lung injury. DESIGN: Prospective, randomized, laboratory study. SETTING: Experimental laboratory at a biomedical institute. SUBJECTS: Twenty male rabbits weighing 2.5-3.5 kg. INTERVENTIONS: Saline or LPS (5 mg/kg of body weight) was administered intravenously with or without NO inhalation (10 ppm) in each group of five rabbits. MEASUREMENTS AND MAIN RESULTS: LPS increased the lung leak index, the neutrophils and NO levels in bronchoalveolar lavage fluid, and NO levels produced by resting and stimulated alveolar macrophages. Inhaled NO decreased the lung leak index, the neutrophils and NO levels as measured by nitrite levels in the lavage fluid, and NO produced by the resting and stimulated alveolar macrophages. Inhaled NO also blocked the activities of reactive oxygen species and nuclear factor-kappaB binding to DNA in lavage cells and in alveolar macrophages. CONCLUSION: Inhaled NO attenuates LPS-induced acute lung injury, possibly by decreasing NO production in the lungs. The mechanism of reducing NO production resulting from inhaled NO may involve, in part, the activities of reactive oxygen species and/or nuclear factor-kappaB. PMID- 11395598 TI - Partial liquid ventilation in severely surfactant-depleted, spontaneously breathing rabbits supported by proportional assist ventilation. AB - OBJECTIVE: We hypothesized that partial liquid ventilation (PLV) would improve oxygenation in nonparalyzed, surfactant-deficient rabbits breathing spontaneously while supported by proportional assist ventilation (PAV). This ventilation mode compensates for low pulmonary compliance and high resistance and thereby facilitates spontaneous breathing. DESIGN: Randomized trial. SETTING: University animal research facility. SUBJECTS: Twenty-six anesthetized New Zealand white rabbits weighing 2592 +/- 237g (mean +/- sd). INTERVENTIONS: After pulmonary lavage (target Pao2 <100 mm Hg on mechanical ventilation with 6 cm H2O of positive end-expiratory pressure [PEEP] and an Fio2 of 1.0), rabbits were randomized to PAV (PEEP of 8 cm H2O) with or without PLV. PLV rabbits received 25 mL/kg of perfluorocarbon by intratracheal infusion (1 mL/kg/min). Pao2, Paco2, tidal volume, respiratory rate, minute ventilation, mean airway pressure, arterial blood pressure, heart rate, pulmonary compliance, and airway resistance were measured. Evaporated perfluorocarbon was refilled every 30 mins in PLV animals. After 5 hrs, animals were killed and lungs were removed. Lung injury was evaluated using a histologic score. MAIN RESULTS: Pao2 and compliance were significantly higher in PLV rabbits compared with controls (p <.05, analysis of variance for repeated measures). All other parameters were similar in both groups. CONCLUSIONS: PLV improved oxygenation and pulmonary compliance in spontaneously breathing, severely surfactant-depleted rabbits supported by PAV. The severity of lung injury by histology was unaffected. PMID- 11395599 TI - Beneficial effect of glycoprotein IIb/IIIa inhibitor (AZ-1) on endothelium in Escherichia coli endotoxin-induced shock. AB - OBJECTIVE: To investigate the effects of AZ-1, a murine monoclonal antiglycoprotein-IIb/IIIa antibody, on endothelium and on hemostasis in a rabbit endotoxic shock model. DESIGN: Prospective laboratory study. SETTING: University laboratory. SUBJECTS: Thirty-five male New-Zealand rabbits. INTERVENTIONS: In vitro vascular reactivity, endothelium CD31-PECAM1 immunohistochemistry, plasma coagulation factors, and monocyte tissue factor determination were performed 1 day and/or 5 days after onset of endotoxic shock (0.5 mg/kg, intravenous bolus,Escherichia coli lipopolysaccharide) with or without treatment by AZ-1 (0.5 mg/kg intravenously) given 1 hr after lipopolysaccharide injection. MEASUREMENTS AND MAIN RESULTS: Metabolic acidosis and coagulation activation confirmed the presence of shock. AZ-1 treatment improved endothelial-dependent relaxation at 1 day (maximal effect = 87.2 +/- 4.0% vs. 60.9 +/- 5.2% in the nontreated group, p <.05) and at 5 days (maximal effect = 84.5 +/- 3.5% vs. 56.6 +/- 8.2% in the nontreated group, p <.05). Endotoxin-induced endothelial injury was decreased significantly by AZ-1 at 1 day (6.4 +/- 1.9% vs. 10.3 +/- 0.8% in the nontreated group, p <.05) and at 5 days (6.3 +/- 2.0% vs. 20.2 +/- 1.2% in the nontreated group, p <.05). Monocyte tissue factor expression was significantly reduced at 5 days. CONCLUSIONS: These data indicate that potent inhibition of platelet function via antiglycoprotein-IIb/IIIa receptor blockade can inhibit coagulation activation and protect against endothelial dysfunction and histologic injury in endotoxin-induced shock. PMID- 11395601 TI - Blockade of central histaminergic H2 receptors aggravates ischemic neuronal damage in gerbil hippocampus. AB - OBJECTIVE: Histaminergic H2 antagonists have been reported to provoke central nervous system dysfunction in humans. They also aggravate ischemic neuronal damage in experimental animals. Because energy failure and glutamate release are crucial factors in ischemic neuronal damage, the effects of ranitidine on energy state and the extracellular concentration of glutamate were investigated in gerbil brain. DESIGN: Prospective, randomized, controlled animal study. SETTING: University animal laboratory. SUBJECTS: Male Mongolian gerbils. INTERVENTIONS: The changes in the direct-current potential shift in the hippocampal CA1 area produced by transient forebrain ischemia for 2.5 mins were compared in gerbils pretreated with saline or ranitidine (10 nmol) intracerebroventricularly. The histologic outcome was evaluated 7 days after ischemia by observing the delayed neuronal death in these animals. In a second study, brain concentrations of adenosine 5'-triphosphate after various durations of decapitation ischemia were determined, and the effect of ranitidine was evaluated. In a third experiment, changes in the extracellular concentrations of excitatory amino acids during forebrain ischemia were examined by a microdialysis procedure. MEASUREMENTS AND MAIN RESULTS: The forebrain ischemia produced a sudden shift in the membrane potential 62 +/- 5 secs (mean +/- sd, n = 6) after the start of ischemia. The preischemic administration of ranitidine facilitated onset of depolarization (38 +/- 8 secs; p <.01). The histologic outcome was aggravated by ranitidine (p <.01). Decapitation ischemia reduced brain adenosine 5'-triphosphate concentration rapidly. Ranitidine facilitated the ischemic reduction in adenosine 5'-triphosphate, and the value after 1 min was 55% of that in the corresponding saline group (p <.01). Ranitidine enhanced the ischemic increase in the glutamate concentration, and the peak value in the ranitidine group was 316% of that in the saline group (p <.05). CONCLUSION: The deleterious effect of ranitidine on ischemic neuronal damage may involve the increase in the extracellular concentration of glutamate and facilitation of energy depletion in an anaerobic state. PMID- 11395602 TI - Nitric oxide inhalation increases alveolar gas exchange by decreasing deadspace volume. AB - OBJECTIVE: To test the hypothesis that nitric oxide inhalation facilitates CO2 elimination by decreasing alveolar deadspace in an ovine model of acute lung injury. DESIGN: Prospective, placebo-controlled, randomized, crossover model. SETTING: University research laboratory. SUBJECTS: Eleven mixed-breed adult sheep. INTERVENTIONS: To induce acute lung injury, hydrochloric acid was instilled into the tracheas of paralyzed sheep receiving controlled mechanical ventilation. Each sheep breathed 0 ppm, 5 ppm, and 20 ppm nitric oxide in random order. MEASUREMENTS AND MAIN RESULTS: Estimates of alveolar deadspace volumes and arterial-to-end tidal CO2 partial pressure differences were used as indicators of CO2 elimination efficiency. At a constant minute ventilation, nitric oxide inhalation caused dose-independent decreases in Paco2 (p <.05), alveolar deadspace (p <.01), and arterial-to-end tidal CO2 partial pressure differences (p <.01). We found that estimates of arterial-to-end tidal CO2 partial pressure differences may be used to predict alveolar deadspace volume (r2 =.86, p <.05). CONCLUSIONS: Estimates of arterial-to-end tidal CO2 partial pressure differences are reliable indicators of alveolar deadspace. Both values decreased during nitric oxide inhalation in our model of acutely injured lungs. This finding supports the idea that nitric oxide inhalation facilitates CO2 elimination in acutely injured lungs. Future studies are needed to determine whether nitric oxide therapy can be used to reduce the work of breathing in selected patients with cardiopulmonary disorders. PMID- 11395604 TI - Glycine attenuates hepatocellular depression during early sepsis and reduces sepsis-induced mortality. AB - OBJECTIVES: To determine whether administration of glycine, a nonessential amino acid, early after the onset of polymicrobial sepsis has any beneficial effects on hepatocellular function and the survivability of septic animals and, if so, whether the beneficial effects of glycine are associated with down-regulation of proinflammatory cytokine tumor necrosis factor-alpha production. DESIGN: Prospective, controlled, and randomized animal study. SETTING: A university research laboratory. SUBJECTS: Male adult rats were subjected to polymicrobial sepsis by cecal ligation and puncture or sham operation followed by the administration of normal saline solution. MEASUREMENTS AND MAIN RESULTS: At 1 hr after cecal ligation and puncture, glycine (0.6 mmol/kg) or vehicle (normal saline solution) was administered intravenously over 15 mins. At 5 hrs after cecal ligation and puncture (i.e., early stage of sepsis), hepatocellular function (i.e., the maximal velocity and efficiency of in vivo indocyanine green clearance) was determined and hepatocyte injury was assessed by measuring plasma concentrations of alpha-gluthathione S-transferase. Serum tumor necrosis factor alpha was measured by enzyme-linked immunosorbent assay. In additional animals, the necrotic cecum was excised at 20 hrs after cecal ligation and puncture, the peritoneal cavity was irrigated with saline, and the midline incision was closed in layers. Mortality was monitored for 10 days thereafter. The results indicate that hepatocellular function was depressed in the early stage of sepsis (i.e., 5 hrs after cecal ligation and puncture) as indicated by significant decreases in both maximal velocity and transport efficiency of in vivo indocyanine green clearance. Plasma concentrations of alpha-gluthathione S-transferase and tumor necrosis factor-alpha were elevated significantly at that interval after cecal ligation and puncture. Administration of glycine 1 hr after cecal ligation and puncture, however, increased maximal velocity and maximal efficiency by 60% and 101% (p <.05), respectively. Glycine administration in septic animals decreased alpha-gluthathione S-transferase and tumor necrosis factor-alpha by 43% and 80% (p <.05). In addition, glycine treatment decreased the mortality rate from 50% to 0% (p <.05) at 10 days after cecal ligation and puncture and cecal excision. CONCLUSIONS: It appears that the beneficial effect of glycine on hepatocyte function and integrity in sepsis may be mediated via down-regulation of tumor necrosis factor-alpha. Because administration of glycine attenuated hepatocellular depression and injury during early sepsis and decreased sepsis induced mortality rates, this amino acid appears to be a useful adjunct for maintaining cellular functions and preventing lethality from polymicrobial sepsis. PMID- 11395606 TI - Effects of the inspiratory pressure waveform during patient-triggered ventilation on pulmonary stretch receptor and phrenic nerve activity in cats. AB - OBJECTIVE: To examine the effects of square wave, sinusoidal, and linear inspiratory pressure waveforms during pressure-controlled assist/control ventilation on the firing pattern of pulmonary stretch receptors and phrenic nerve activity. DESIGN: Experimental, comparative study. SETTING: Research laboratory at a university biomedical center. SUBJECTS: Nine anesthetized, endotracheally intubated young cats (2.5-3.4 kg). INTERVENTION: With interposed periods of continuous positive airway pressure (0.2 kPa), each cat was exposed to periods of assist/control ventilation with three different pressure waveforms, where the peak inspiratory pressure (0.74 +/- 0.13 kPa), end-expiratory pressure (0.2 +/- 0.02 kPa), and tidal volume (14.9 +/- 5.22 mL/kg) were kept constant. Preset controlled ventilator rate was set below the rate of spontaneous breathing, and the mechanical inflation time equaled the inspiratory time during spontaneous breathing on continuous positive airway pressure. MEASUREMENTS AND MAIN RESULTS: Respiratory rate and arterial blood gases did not change between the three pressure waveforms during assist/control ventilation. Peak pulmonary stretch receptor activity was lower and mean phrenic nerve activity higher during continuous positive airway pressure than during assist/control ventilation (p <.05). Peak inspiratory pulmonary stretch receptor activity was the same with all three pressure waveforms (82 +/- 17 impulses.sec-1) but occurred earlier with square wave than with sinusoidal or linear pressure waveforms (p <.05). The total number of impulses in the phrenic nerve activity burst was smaller with square wave than with the other two pressure waveforms (0.21 +/- 0.17 vs. 0.33 +/- 0.27 and 0.42 +/- 0.30 arbitrary units; p <.05), and the phrenic nerve activity burst duration was shorter with square wave (1.10 +/- 0.45 vs. 1.54 +/- 0.36 and 1.64 +/- 0.25 secs; p <.05). CONCLUSION: Square wave pressure waveform during pressure controlled assist/control ventilation strongly inhibits spontaneous inspiratory activity in cats. One mechanism for this inhibition is earlier and sustained peak pulmonary stretch receptor activity during inspiration. These findings show that differences in inspiratory pressure waveforms influence the spontaneous breathing effort during assist/control ventilation in cats. PMID- 11395607 TI - ONO-1714, a new inducible nitric oxide synthase inhibitor, attenuates diaphragmatic dysfunction associated with cerulein-induced pancreatitis in rats. AB - OBJECTIVES: Acute experimental pancreatitis (induced by cerulein) recently has been reported to cause marked diaphragmatic dysfunction, which may contribute to respiratory distress in this setting. In cerulein-induced acute pancreatitis, expression of inducible nitric oxide synthase is induced to produce a large amount of nitric oxide. Nitric oxide excessively produced has been implicated in diaphragmatic dysfunction induced by a variety of etiologies. The aims of the current study were, first, to examine whether nitric oxide overproduced through inducible nitric oxide synthase is involved in cerulein-induced impairment of diaphragmatic function, and second, if demonstrated, to assess effects of ONO 1714, an inducible nitric oxide synthase inhibitor, on diaphragmatic dysfunction associated with cerulein-induced acute pancreatitis. DESIGN: Prospective, randomized animal study. SETTING: University research laboratory. SUBJECTS: Ninety-one male Sprague-Dawley rats, weighing 200-250 g. INTERVENTIONS: Rats were randomly divided into seven groups (n = 8 each): CONT-SAL, CAER-SAL, CONT-ONO, CAER-DEX, CAER-AMI, CAER-ONOhigh, and CAER-ONOlow. Groups labeled CAER received two consecutive intraperitoneal doses (50 microg/kg) of cerulein, whereas groups labeled CONT received two consecutive intraperitoneal injections of saline. Groups labeled SAL received intraperitoneal saline before cerulein or saline. The group labeled DEX received 2 mg/kg intraperitoneal dexamethasone, and the group labeled AMI received 100 mg/kg intraperitoneal aminoguanidine. The groups labeled ONO, ONOhigh, and ONOlow received ONO-1714 at 0.1 mg/kg, 0.1 mg/kg, and 0.03 mg/kg, respectively, before cerulein or saline. MEASUREMENTS AND MAIN RESULTS: Diaphragmatic contractility and fatigability were assessed in vitro by using muscle strips excised from the costal diaphragms 6 hrs after the first dose of cerulein or saline. Expression of inducible nitric oxide synthase protein in the diaphragm was assessed by immunohistochemistry by using anti-inducible nitric oxide synthase antibody. Plasma concentrations of nitrite plus nitrate and diaphragmatic concentrations of malondialdehyde were measured. With another set of rats (n = 5 each group), diaphragmatic inducible nitric oxide synthase activity was determined. Twitch and tetanic tensions and tensions generated during fatigue trial were lower in group CAER-SAL than in group CONT-SAL. Cerulein increased diaphragmatic malondialdehyde and plasma nitrite plus nitrate concentrations. Positive immunostaining for inducible nitric oxide synthase protein was found in group CAER-SAL. Dexamethasone and aminoguanidine attenuated the diaphragmatic mechanical damages. A high dose of ONO-1714 attenuated cerulein induced impairment of diaphragmatic contractility and endurance capacity, although a low dose of the drug failed to do so. CONCLUSIONS: Cerulein-induced diaphragmatic dysfunction was attributable, in part, to nitric oxide overproduced via inducible nitric oxide synthase. Pretreatment with ONO-1714 at a dose of 0.1 mg/kg attenuated diaphragmatic dysfunction associated with cerulein-induced pancreatitis in rats assessed by contractile profiles and endurance capacity. This beneficial effect of ONO-1714 may be attributable, in part, to inhibition of diaphragmatic lipid peroxidation induced by nitric oxide-derived free radicals. PMID- 11395608 TI - Role of central nervous system nitric oxide in the development of neurogenic pulmonary edema in rats. AB - OBJECTIVE: The present study was undertaken to evaluate roles of nitric oxide in the central nervous system in the development of neurogenic pulmonary edema. Nitric oxide donor compounds have been reported to be effective for controlling some kinds of pulmonary edema. DESIGN: Randomized trial. SETTING: Experimental university pharmacology laboratory. SUBJECTS: Wistar rats anesthetized with pentobarbital. INTERVENTIONS: Neurogenic pulmonary edema was induced by injections of fibrinogen and thrombin into the cisterna magna. Physiologic roles of nitric oxide were evaluated by using NG-nitro-l-arginine methyl ester (a nitric oxide synthase inhibitor) or l-arginine (a nitric oxide donor compound). Vagus nerves were either left intact or bilaterally severed 20 mins before the injections of fibrinogen and thrombin. MEASUREMENTS AND MAIN RESULTS: Because enhanced sympathetic nerve activity mediates neurogenic pulmonary edema, the concentration of neuropeptide Y, a neurotransmitter, in edema fluid was measured by using enzyme-linked immunosorbent assay. To evaluate the severity of pulmonary edema and pulmonary vascular permeability, lung water content and protein concentration in edema fluid were analyzed. In rats with intact vagus nerves, injection of NG-nitro-l-arginine methyl ester into the cisterna magna worsened the pulmonary edema, whereas l-arginine had no effect. In contrast, in vagotomized rats, l-arginine abrogated pulmonary edema, whereas NG-nitro-l arginine methyl ester exerted no influence. Likewise, the ratio of edema fluid protein to serum protein and the neuropeptide Y concentration were increased by NG-nitro-l-arginine methyl ester in rats with the vagus nerves intact and were diminished by l-arginine in vagotomized rats. CONCLUSIONS: Neurogenic pulmonary edema is characterized by elevated pulmonary vascular permeability and may be inhibited by nitric oxide production in the medulla oblongata. PMID- 11395610 TI - Role of protein kinase C and phosphatases in the pulmonary vasculature of neonatal piglets. AB - OBJECTIVE: Persistent pulmonary hypertension of the newborn is characterized by the presence of intense vasoconstriction and vascular remodeling. Protein tyrosine phosphorylation has been recognized as a critical regulatory element in signal transduction, because it is dynamically regulated by the opposing actions of protein tyrosine kinases and protein tyrosine phosphatases. The objectives of this study were to investigate the role of protein kinase C and phosphatases in the neonatal pulmonary vasculature of normoxic and chronically hypoxic piglets. DESIGN: Prospective, randomized, unblinded study. SETTING: Hospital research laboratory. SUBJECTS: Newborn Yorkshire-Landrace piglets. INTERVENTIONS: Normoxic animals were 3-6 days old. Hypoxic animals were exposed to hypoxia (Fio2 0.10) between 1 and 15 days of age to induce pulmonary hypertension and then were studied. MEASUREMENTS AND MAIN RESULTS: In isolated perfused lungs from normoxic piglets, we measured the perfusion pressure to assess the vasoconstrictor response to protein kinase C activation with phorbol 12,13-dibutyrate or 1-oleyl 2-acetyl-glycerol. We also assessed the effect of protein kinase C inhibition with staurosporine (2 x 10-6M) and chelerythrine (5 x 10-5M) on endothelin-1 induced pulmonary vasoconstriction. We then examined the effect of chelerythrine and phosphatase inhibition with phenylarsine oxide on the baseline perfusion pressure of normoxic and chronically hypoxic piglets. Phorbol 12,13-dibutyrate and 1-oleyl-2-acetyl-glycerol caused a sustained, dose-dependent increase in perfusion pressure, with relative potencies about 100- and 1000-fold less than endothelin-1, respectively. Protein kinase C inhibitors, chelerythrine and staurosporine, decreased the constrictor response to endothelin-1. Chelerythrine did not affect baseline perfusion pressure in the normoxic animal, whereas it lowered pulmonary vascular tone in chronically hypoxic animals. With respect to phosphatases, phenylarsine oxide significantly increased perfusion pressure in normoxia as well as in hypoxia. CONCLUSIONS: These findings confirm that protein kinase C activation causes sustained vasoconstriction in the neonatal pulmonary vasculature and mediates the vasoconstrictor action of potent peptides, like endothelin-1. These findings also confirm that protein kinase C activation could be induced by hypoxic exposure in the neonatal piglet pulmonary vasculature. Phosphatases appear to modulate pulmonary vascular tone in the normoxic and hypoxic newborn piglet. PMID- 11395611 TI - Growth hormone/insulin-like growth factors axis in children undergoing cardiac surgery. AB - OBJECTIVES: To describe the sequential changes in the growth hormone (GH)/insulin like growth factors axis and their relationship with nitrogen balance in children following cardiac surgery. DESIGN: Prospective, descriptive study. SETTING: Pediatric intensive care unit of a university hospital. PATIENTS: Twenty three postoperative cardiac surgical patients after bypass. INTERVENTIONS: Blood and urine samples were taken on days 1, 2, and 7 of pediatric intensive care unit admission. An intraanesthesia, presurgery sample was also obtained. MEASUREMENTS AND MAIN RESULTS: Serum concentrations of insulin, insulin growth factor-I (IGF I), insulin growth factor binding proteins 1 and 3 (IGFBP-1 and IGFBP-3), growth hormone binding protein (GHBP), and urinary concentrations of GH and free cortisol (UFC) were measured on days 1, 2, and 7 of the study period. C-reactive protein and prealbumin, were also measured in blood samples as conventional markers of inflammatory or nutritional status. Pediatric Risk of Mortality II score and UFC were used as indicators of acute stress. The nitrogen balance and urinary nitrogen urea excretion were used as markers of catabolic state. Urinary concentrations of GH were high from days 1 to 7. Plasma concentrations of IGF-I and GHBP were low and remained low throughout the study period. IGFBP-3 levels were below normal but without reaching statistical significance. The IGFBP-1 levels were initially high but descended progressively toward normal values. Urinary nitrogen urea production was persistently elevated and was associated with a negative nitrogen balance. No relationship was found between nitrogen balance and IGF-I, prealbumin, or C-reactive protein. CONCLUSIONS: A GH-resistant state is observed in postoperative children following cardiac surgery. Stress response is characterized by an elevation of growth hormone secretion that is not followed by the corresponding increment in IGF-I and IGFBP-3 concentrations. These hormonal changes may be permissive for the catabolic state of these patients. IGF-I and IGFBP-1 and -3 are not related to either nitrogen balance or urinary nitrogen urea excretion. PMID- 11395615 TI - Open lung biopsy in children with respiratory failure. AB - OBJECTIVE: To evaluate benefits and risks of open lung biopsy in children with respiratory failure. DESIGN: Retrospective chart review. SETTING: A 36-bed pediatric critical care unit in a tertiary care, university-based hospital. PATIENTS: We studied 31 patients with respiratory failure who underwent 33 open lung biopsies. MEASUREMENTS AND MAIN RESULTS: The charts of all children in the critical care unit with respiratory failure who underwent an open lung biopsy over a 10-yr period (1989-98) were reviewed. Of 33 open lung biopsies performed, 76% (25 of 33) led to a relevant change in medical management. Complications were seen in 45% of patients, predominantly attributable to airleak (33%) without affecting respiratory function. An infectious agent was detected by open lung biopsy in ten patients; bronchoalveolar lavage performed before open lung biopsy failed to isolate the infection in eight of ten patients. CONCLUSIONS: In children with undiagnosed or persisting respiratory failure, open lung biopsy is a useful diagnostic procedure that leads to significant changes in medical management and increases the diagnostic yield for infections. Despite the relatively high complication rate, open lung biopsy should be performed routinely in this group of patients. PMID- 11395613 TI - Can generic scores (Pediatric Risk of Mortality and Pediatric Index of Mortality) replace specific scores in predicting the outcome of presumed meningococcal septic shock in children? AB - OBJECTIVE: To compare, in children with septic shock and purpura, the accuracy in predicting death of two specific scores (the MenOPP bedside clinical [MOC] score of Gedde Dahl and the score of Groupe Francophone de Reanimation Pediatrique [GFRP]), the C-reactive protein (CRP) level, and the two pediatric generic scores (the Pediatric Risk of Mortality [PRISM] and Pediatric Index of Mortality [PIM] scores). DESIGN: Prospective, population-based study with analysis of previous comparative studies. SETTING: A 14-bed pediatric intensive care unit in a university hospital. PATIENTS: All children admitted consecutively to the pediatric intensive care unit with septic shock and purpura (n = 58, with 16 deaths [27.6%]) from January 1993 to May 2000. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The MOC and GFRP scores and the CRP level were prospectively determined at admission. The PRISM score was prospectively calculated within 24 hrs of admission or at the time of death, and the PIM score was calculated retrospectively between 1993 and 1997 and then prospectively from admission data. The nonparametric estimate of the area under the receiver operating characteristic curves (AUC) was calculated from the raw data using the Wilcoxon Mann-Whitney two-sample statistic, and the standard error of the AUCs was calculated with DeLong's method. All the scores had an AUC >0.80, the PRISM probability of death having the best one (0.96 +/- 0.02). The PRISM value, which is easier to calculate, had an AUC of 0.95 +/- 0.02. The PRISM score performed significantly better than the PIM score (AUC, 0.83 +/- 0.06; p <.01) and the CRP level (AUC, 0.80 +/- 0.06; p <.01); however, there was no significant difference between the MOC (AUC, 0.91 +/- 0.04) and GFRP scores (AUC, 0.87 +/- 0.05). Analyzing literature and calculating AUCs from original data of previous studies, we observed that the superiority of the PRISM score had never been demonstrated in meningococcal diseases. CONCLUSIONS: The PRISM score performed better than the PIM score, and was not surpassed by specific scores. Thus, we propose its use for outcome prediction in children with septic shock and purpura. However, if the PRISM score is to be used as inclusion criterion for clinical trials, it should be evaluated within a few hours after admission. PMID- 11395616 TI - Percutaneous dilational tracheostomy: a comparison of single- versus multiple dilator techniques. AB - OBJECTIVE: To compare the safety and efficacy of single- vs. multiple-dilator techniques in the performance of percutaneous dilational tracheostomy. DESIGN: Prospective randomized trial. SETTING: Intensive care units at a level 1 trauma center. PATIENTS: Fifty consecutive patients requiring tracheostomy for airway control or prolonged mechanical ventilatory support. INTERVENTIONS: Patients were randomized to receive a percutaneous dilational tracheostomy by either the single or multiple-dilator technique described by Ciaglia. MEASUREMENTS AND MAIN RESULTS: Percutaneous dilational tracheostomy was performed using the single dilator technique in 6:01 +/- 3:03 mins and by the multiple-dilator technique in 10:01 +/- 4:26 mins (p <.0006). There were no statistically significant differences in complication rates between the two techniques. No major complications occurred with either technique. CONCLUSION: The single-dilator percutaneous tracheostomy technique is a safe, cost-effective, and more rapidly performed method for bedside tracheostomy in the intensive care unit. PMID- 11395617 TI - Mechanistic scheme and effect of "extended sigh" as a recruitment maneuver in patients with acute respiratory distress syndrome: a preliminary study. AB - OBJECTIVE: To devise a new form of sigh ("extended sigh") capable of providing a sufficient recruiting pressure x time, and to test it as a recruitment maneuver in patients with acute respiratory distress syndrome. DESIGN: Prospective uncontrolled clinical trial. SETTING: Medical intensive care unit of a university affiliated hospital. PATIENTS: Twenty consecutive patients diagnosed with acute respiratory distress syndrome (18 men, 2 women, age 59 +/- 10 yrs). INTERVENTIONS: From baseline settings of tidal volume (Vt) 8 mL/kg and positive end-expiratory pressure (PEEP) 10 cm H2O on volume control mode with the high pressure limit at 40 cm H2O, the Vt-PEEP values were changed to 6-15, 4-20, and 2 25, each step being 30 secs (inflation phase). After Vt-PEEP 2-25, the mode was switched to continuous positive airway pressure of 30 cm H2O for a duration of 30 secs (pause), after which the baseline setting was resumed following the reverse sequence of inflation (deflation phase). This extended sigh was performed twice with 1 min of baseline ventilation between. MEASUREMENTS AND RESULTS: Airway pressures and hemodynamic parameters were traced at each step during the extended sigh. Arterial blood gases and physiologic parameters were determined before the extended sigh (pre-extended sigh), at 5 mins after two extended sighs (post extended sigh), and then every 15 mins for 1 hr. In our average patient, the recruiting pressure x time of the inflation phase was estimated to be 32.8-35.4 cm H2O x 90 secs. Compared with the inflation phase, inspiratory pause pressure of the deflation phase was lower at Vt-PEEP 6-15 (28.9 +/- 2.7 cm H2O vs. 27.3 +/ 2.8 cm H2O) and 4-20 (31.8 +/- 2.9 cm H2O vs. 31.1 +/- 2.9 cm H2O; both p <.05). Compared with pre-extended sigh, Pao2 (81.5 +/- 15.3 mm Hg vs. 104.8 +/- 25.0 mm Hg; p <.001) and static respiratory compliance both increased post-extended sigh (27.9 +/- 7.9 mL/cm H2O vs. 30.2 +/- 9.7 mL/cm H2O; p =.009). Improvement in these parameters was sustained above pre-extended sigh for the duration of the study. Major hemodynamic or respiratory complications were not noted during the study. CONCLUSION: We present a new form of sigh (i.e., extended sigh) capable of achieving an augmented recruiting pressure x time through a prolonged inflation on a gradually increased end-expiratory pressure. In view of the sustained effect and absence of major complications in our patients, extended sigh could be a useful recruitment maneuver in acute respiratory distress syndrome. PMID- 11395618 TI - Effects of different plasma substitutes on blood coagulation: a comparative review. AB - OBJECTIVE: To compare the effects of different colloid plasma substitutes on blood coagulation and postoperative blood loss. DATA SOURCES: Relevant studies were obtained from the medical literature. STUDY SELECTION: Articles were selected that provided data on the effects of colloids on hemostasis and postoperative blood loss in humans. Studies comparing different colloids were looked for using MEDLINE and by searching through the references of studies as they were collected. DATA SYNTHESIS: Articles were reviewed and relevant data were extracted and partly presented in comparative tables. CONCLUSIONS: Dextran, gelatin, and hydroxyethyl starch (HES) all can induce a specific decrease of von Willebrand factor and factor VIII:c. Blood coagulation is most impaired by dextran and high molecular weight HES, both associated with increased postoperative blood loss. The effects of HES on blood coagulation have been shown to depend on its molecular weight and rate of elimination. Detrimental effects have been shown for high molecular weight HES. Medium molecular weight (MMW)-HES with a high degree of substitution (HES 200/0.62) and MMW-HES with high C2/C6 hydroxyethylation ratio (HES 200/0.5/13) are slowly degradable and have been shown to impair blood coagulation after repeated administration. Rapidly degradable HES 200/0.5/6 and gelatin-based plasma expanders appear not to impair hemostasis. However, based on the reviewed literature, all artificial colloids could potentially induce increased bleeding tendency after infusion of very large volumes and especially when given to patients with even mild forms of von Willebrand disease. In those circumstances, crystalloid solutions or alternatives such as plasma or albumin, although associated with other serious complications, could be considered. PMID- 11395620 TI - Rational approaches to antibiotic therapy of ventilator-associated pneumonias. PMID- 11395619 TI - Quantifying learning in medical students during a critical care medicine elective: a comparison of three evaluation instruments. AB - OBJECTIVE: To compare three different evaluative instruments and determine which is able to measure different aspects of medical student learning. DESIGN: Student learning was evaluated by using written examinations, objective structured clinical examination, and patient simulator that used two clinical scenarios before and after a structured critical care elective, by using a crossover design. PARTICIPATION: Twenty-four 4th-yr students enrolled in the critical care medicine elective. INTERVENTIONS: All students took a multiple-choice written examination; evaluated a live simulated critically ill patient, requested data from a nurse, and intervened as appropriate at different stations (objective structured clinical examination); and evaluated the computer-controlled patient simulator and intervened as appropriate. MEASUREMENTS AND MAIN RESULTS: Students' knowledge was assessed by using a multiple-choice examination containing the same data incorporated into the other examinations. Student performance on the objective structured clinical examination was evaluated at five stations. Both objective structured clinical examination and simulator tests were videotaped for subsequent scores of responses, quality of responses, and response time. The videotapes were reviewed for specific behaviors by faculty masked to time of examination. Students were expected to perform the following: a) assess airway, breathing, and circulation; b) prepare a mannequin for intubation; c) provide appropriate ventilator settings; d) manage hypotension; and e) request, interpret, and provide appropriate intervention for pulmonary artery catheter data. Students were expected to perform identical behaviors during the simulator examination; however, the entire examination was performed on the whole-body computer-controlled mannequin. The primary outcome measure was the difference in examination scores before and after the rotation. The mean preelective scores were 77 +/- 16%, 47 +/- 15%, and 41 +/- 14% for the written examination, objective structured clinical examination, and simulator, respectively, compared with 89 +/- 11%, 76 +/- 12%, and 62 +/- 15% after the elective (p <.0001). Prerotation scores for the written examination were significantly higher than the objective structured clinical examination or the simulator; postrotation scores were highest for the written examination and lowest for the simulator. CONCLUSION: Written examinations measure acquisition of knowledge but fail to predict if students can apply knowledge to problem solving, whereas both the objective structured clinical examination and the computer-controlled patient simulator can be used as effective performance evaluation tools. PMID- 11395621 TI - Does multiple trauma increase the mortality rate from severe traumatic brain injury by increasing the burden of secondary cerebral ischemic insults? PMID- 11395623 TI - To just say NO or I don't inhale? PMID- 11395622 TI - Correlating coronary angiographic stenosis morphology with clinical presentation and troponin levels in acute coronary syndromes: a contrarian's hypothesis. PMID- 11395624 TI - IIb/IIIa or not IIb/IIIa, that is adhesion. PMID- 11395625 TI - Measurement of the alveolar deadspace: are we there yet? PMID- 11395626 TI - Percutaneous tracheostomy: what technique is it? PMID- 11395627 TI - Assessing medical competence--virtual reality? PMID- 11395628 TI - Improved outcomes of children with malignancy admitted to a pediatric intensive care. PMID- 11395629 TI - A strategy to avoid respiratory motion artifact in mechanically ventilated patients undergoing helical chest computed tomography angiography. PMID- 11395630 TI - High-frequency oscillation and partial liquid ventilation. PMID- 11395631 TI - Gastric emptying in the critically ill. PMID- 11395633 TI - Hierarchy, Tolerance, and Dominance in the Antitumor T-Cell Response. PMID- 11395634 TI - MHC Class II-Restricted Tumor Antigens Recognized by CD4+ T Cells: New Strategies for Cancer Vaccine Design. AB - SUMMARY: The adoptive transfer of tumor-infiltrating lymphocytes (TIL) can mediate tumor regression in patients with melanoma. This finding has led to the identification and characterization of tumor-associated antigens recognized by CD8+ TIL. Several clinical trials based on the genes recognized by these CD8+ T cells have been attempted, but with only limited success. Meanwhile, increasing evidence has demonstrated that CD4+ T cells play important roles in generating and maintaining antitumor immune responses in animal models. These data suggest that it may be necessary to engage both CD4+ and CD8+ T cells for more effective antitumor immunotherapy. In this report, we review emerging molecular approaches in cloning major histocompatibility complex (MHC) class II restricted tumor antigens recognized by CD4+ T cells as well as approaches to identify new MHC class II-restricted epitopes from known tumor antigens recognized by CD8+ cytotoxic T lymphocytes and/or antibodies. Progress made in this field has shed light on the roles of tumor antigen-specific CD4+ T cells in humans; it has also provided new insights into the understanding of tumor genesis and the interaction between tumor and the immune system. More importantly, the discovery of MHC class II-restricted tumor antigens has provided opportunities for developing a new generation of cancer vaccines aimed at eliciting both CD4+ and CD8+ T-cell responses against tumor. PMID- 11395635 TI - Antitumor Effect on Murine Renal Cell Carcinoma by Autologous Tumor Vaccines Genetically Modified with Granulocyte-Macrophage Colony-Stimulating Factor and Interleukin-6 Cells. AB - SUMMARY: The authors evaluted the efficacy of vaccination with murine renal cell carcinoma (Renca) secreting the granulocyte-macrophage colony-stimulating factor (GM-CSF) gene and interleukin-6 (IL-6) gene for the treatment of Renca tumor. Murine GM-CSF and murine IL-6 genes were introduced and expressed in Renca cells (Renca-GM-CSF and Renca-IL-6). For a prevaccination study, wild-type Renca cells were injected subcutaneously into Balb/c mice that had been vaccinated three times with inactivated wild-type Renca, Renca-GM-CSF, Renca-IL-6, or a mixture of Renca-GM-CSF and Renca-IL-6 cells 7, 14, and 21 days before this tumor inoculation. For vaccination experiments, Renca tumor-bearing (8 to 10 mm) mice were injected subcutaneously weekly for 3 weeks with inactivated wild-type Renca cells, or either one or a combination of Renca-GM-CSF and Renca-IL-6. A nonvaccinated control was included in all experiments. The animals were monitored for survival and tumor development for 8 weeks. Mice inoculated with wild-type Renca alone died from the tumor within 35 days. Renca-IL-6 grew slower than wild type Renca (p < 0.05). No tumor was produced by Renca-GM-CSF. Prevaccination with the combination of Renca-GM-CSF and Renca-IL-6 prevented subsequently inoculated wild-type Renca from forming tumors, and prevaccination with either one of them, compared with prevaccination with wild-type Renca, retarded tumor growth and prolonged survival time. Tumor-bearing mice vaccinated with wild-type Renca died within 42 days. Vaccination with Renca-GM-CSF or Renca-IL-6 alone prolonged the survival time, but only Renca-GM-CSF drastically reduced the tumor size. Vaccination with the combination of them achieved complete remission. Neither of the cytokine-secreting cells enhanced the expression of MHC class I or II molecules. Autologous tumor cell vaccine secreting GM-CSF is effective in preventing and treating established tumors. Its efficacy is enhanced by the cosecretion of IL-6. PMID- 11395636 TI - Identification of a New Shared HLA-A2.1 Restricted Epitope From the Melanoma Antigen Tyrosinase. AB - SUMMARY: Tyrosinase has many advantages as a target antigen for the immunotherapy of patients with melanoma because it is expressed in nearly all melanoma specimens with a high degree of cellular homogeneity, and its distribution in normal tissues is limited to melanocytes. To broaden our ability to direct cellular immune responses against this protein, we pursued an investigation to identify new shared human leukocyte antigen (HLA)-A2.1 restricted epitopes from tyrosinase. Peptides were synthesized that fit a permissive HLA-A2.1 binding motif and did not span common sites of polymorphism. The binding affinity of each peptide to HLA-A2.1 relative to a standard peptide with intermediate binding affinity was evaluated in a competitive inhibition assay. Twelve peptides were selected that had binding affinities within 80% of that of the standard peptide, and these were used to stimulate peripheral blood mononuclear cells (PBMC) in vitro from three HLA-A2.1+ patients with metastatic melanoma. Cytotoxic T lymphocytes that specifically recognized peptide-pulsed target cells as well as HLA-A2.1+ tyrosinase+ melanoma cells were raised from one patient with tyrosinase:8-17 (CLLWSFQTSA). To evaluate further the immunogenicity of this peptide, PBMC from 23 HLA-A2.1+ patients were stimulated in vitro with tyrosinase:8-17. Eleven bulk T-cell cultures demonstrated specific peptide recognition, and six of these also recognized HLA-A2.1+ tyrosinase+ melanoma cells. These data suggest that tyrosinase:8-17 may be clinically useful for the treatment of patients with melanoma. PMID- 11395637 TI - Polyclonal Antibodies Against gp185HER2 Peptides: Their Putative Role in the Identification of a Particular HER2 Status in Patients With Breast Cancer. AB - SUMMARY: The HER2 oncogene and its relative oncoprotein, gp185HER2, a transmembrane glycoprotein belonging to the epidermal growth factor receptor family, are overexpressed in a wide range of solid tumors including breast and ovarian cancer. In patients with breast cancer, both humoral and cell-mediated HER2 immune responses have been found as well as in some patients with gp185HER2 nonoverexpressing tumors. To establish whether peptide sequences identified as HLA-A2-restricted T-cell epitopes are expressed in breast tumor cell lines and tissues, we produced and characterized by different methodologic approaches polyclonal antibodies raised against four gp185HER2 peptides. Two of the antibodies recognized peptides eluted from the HLA-A2 groove of the mDAmB231 breast cancer cell line expressing a basal level of gp185HER2. Paraffin-embedded primary and metastatic breast tumors were specifically immunostained by all four reagents, thereby showing an overlapping reactivity. When this immunoreactivity was compared with that obtained using two different monoclonal antibodies, in 105 breast primary tumors and 36 corresponding lymph node metastases, we identified a subset of tumors that were negative with anti-gp185HER2 monoclonal antibodies and positive with the four antipeptide antibodies. Our novel observations provide in vivo evidence of the complexity involved in evaluating HER2 expression, and open a new path for understanding the biologic significance of HER2 status in breast tumors. PMID- 11395638 TI - Equipotent Generation of Protective Antitumor Immunity by Various Methods of Dendritic Cell Loading With Whole Cell Tumor Antigens. AB - Multiple clinically applicable methods have been used to induce dendritic cells (DCs) to express whole cell tumor antigens, including pulsing DCs with tumor lysate, and mixing DCs with apoptotic or live tumor cells. Herein we demonstrate, using two different tumor systems, that these methods are equipotent inducers of systemic antitumor immunity. Furthermore, tumor lysate pulsed DC vaccines generate more potent antitumor immunity than immunization with irradiated tumor cells plus the classic adjuvant, Corynebacterium parvum. PMID- 11395639 TI - Avoiding Tolerance Against Prostatic Antigens With Subdominant Peptide Epitopes. AB - SUMMARY: A potential novel therapy for prostate cancer is the induction of immune responses to normal prostate-associated antigens (PAA). One approach is to use synthetic peptides from PAA to educate T cells as a means of developing a defined and specific immunotherapy for prostate cancer. A likely major hurdle when using normal PAA for this type of therapy is the tolerance that the immune system may already have for PAA. To evaluate mechanisms for overcoming tolerance, the authors assessed the level of tolerance to SV40T antigen in a transgenic mouse. The SV40T antigen is selectively expressed in the prostates of mice from the transgenic adenocarcinoma mouse prostate (TRAMP) model. The authors have shown that TRAMP mice are tolerant to a dominant cytotoxic T-lymphocyte (CTL) epitope from the SV40T antigen compared with nontransgenic littermates. The tolerance was exhibited as early as 4 weeks and as late as 24 weeks. The use of multiple injections of an oligonucleotide that contains an unmethylated CpG induced high levels of hematopoiesis but did not overcome the tolerance. Injection of an antibody to activate CD40 increased the CTL response in normal mice but also did not overcome tolerance. However, tolerance in the TRAMP mice was avoided when an epitope that had previously been characterized as a subdominant epitope was administered. The authors are investigating the potential of subdominant epitopes to induce prostatitis and antitumor responses. The results of this work should facilitate the development of immune-based therapies for prostate cancer. PMID- 11395640 TI - Generation of Cytotoxic T Lymphocytes Specific for Human Cytomegalovirus Using Dendritic Cells In Vitro. AB - SUMMARY: For the adoptive immunotherapy in immunodeficient bone marrow transplant recipients to prevent and treat human cytomegalovirus (HCMV)-associated diseases, HCMV-pulsed dendritic cells (DCs) were used as antigen-presenting cells for the induction of cytotoxic T lymphocytes (CTLs) specific to HCMV antigens in vitro. The antiviral CTL responses induced by HCMV-pulsed DCs were as highly efficient as those induced by HCMV-infected dermal fibroblasts, and endogenous viral gene expression was not required to induce virus-specific T-cell lines. The strong cytotoxic activity against HCMV-pp65, known as HCMV major antigen, was identified using autologous B lymphoblastoid cell line expressing pp65 antigen. The cytotoxic activity toward HCMV-infected target cells was found to be mediated primarily by CD8+ T cells, although both CD8+ cells and CD4+ cells were able to lyse autologous virus-infected target cells. The CTLs contained a mixture of effector cells that recognized virus peptides in the context of major histocompatibility complex. This system may be useful for defining the cellular immune response to HCMV and for the treatment of HCMV infection in immunocompromised patients. PMID- 11395641 TI - Meta-analysis of Adjuvant Immunochemotherapy Using OK-432 in Patients With Resected Non-Small-Cell Lung Cancer. AB - SUMMARY: The benefits of immunochemotherapy with a penicillin-treated, lyophilized preparation of Streptococcus pyogenes, OK-432 (Picibanil), were reassessed in patients with resected non-small-cell lung cancer through a meta analysis based on data from 1,520 patients enrolled in 11 randomized clinical trials. All 11 trials were started before 1991, and the subjects had been followed up for at least 5 years after surgery and randomization. In these trials, standard chemotherapy was compared with the same therapy plus OK-432. The endpoint of interest was overall survival, and analysis was based on intent-to treat population without patient exclusion. Data were analyzed using the Mantel Haenszel method. The 5-year survival rate for all eligible patients in the 11 trials was 51.2% in the immunochemotherapy group versus 43.7% in the chemotherapy group. The odds ratio (OR) for overall survival was 0.70 (95% CI = 0.56-0.87, p = 0.0010). Analysis of four trials in which central randomization was performed also reconfirmed a significantly longer survival time for the immunochemotherapy group (OR = 0.66, 95% CI = 0.44-1.00, p = 0.049). Based on these results of meta analysis, it is postulated that postoperative adjuvant immunochemotherapy using OK-432 might improve the survival of patients after resection of non-small-cell lung cancer. PMID- 11395642 TI - Sequential Administration of Interferon-gamma, GM-CSF, and Interleukin-2 in Patients With Metastatic Renal Cell Carcinoma: Results of a Phase II Trial. AB - SUMMARY: Various cytokine combinations have been tested for efficacy in the treatment of metastatic renal cell carcinoma (MRCC). Because several immunologic synergisms between granulocyte-macrophage colony-stimulating-factor (GM-CSF) and interleukin-2 (IL-2) have been demonstrated, this phase II trial was conducted on the efficacy and toxicity of subcutaneous, sequentially administered, interferon gamma (IFNgamma), GM-CSF, and IL-2. Fifty-five consecutive patients with MRCC were treated with 100 &mgr;g recombinant IFNgamma1b administered thrice weekly during weeks 1 and 4, followed by 400 &mgr;g GM-CSF on 5 consecutive days during weeks 2 and 5. In weeks 3 and 6, patients received 4.5 MU recombinant IL-2 from days 1 to 4. The treatment was repeated every 8 weeks. Five (10%) of patients experienced an objective response (complete response [CR]: 2%, partial response [PR]: 8%). Fourteen (26%) patients had stable disease with a median duration of 19 months (6-47+). The median overall survival was 12 months (range: 0.3-44 months). No toxicity greater than World Health Organization grade II was observed, with fever (43%) and erythema (43%) being the most frequent side effects. Compared with other phase II trials with IFNgamma and IL-2 alone, the addition of GM-CSF failed to improve response or survival in patients with MRCC. PMID- 11395643 TI - Tumor Cell Expression of CD59 Is Associated With Resistance to CD20 Serotherapy in Patients With B-Cell Malignancies. AB - SUMMARY: The anti-CD20 chimeric monoclonal antibody rituximab (Rituxan) is used to treat patients with various B-cell tumors, including patients with plasma cell dyscrasias who have CD20+ disease. Many patients with CD20+ disease have either primary unresponsive disease or progress after initially responding to rituximab; therefore, understanding how tumor cells are, or become, resistant to rituximab is of clinical relevance. In this report, we determined whether tumor cells express antigens that block complement-mediated lysis or antibody-dependent cell mediated cytotoxicity (ADCC) and thereby contribute to rituximab resistance. We demonstrate that expression of the complement regulator CD59 is associated with resistance to rituximab-mediated complement lysis of multiple myeloma (MM) and non-Hodgkin's lymphoma (NHL) cell lines. Moreover, neutralization of CD59 using a blocking monoclonal antibody reversed resistance to rituximab-mediated complement lysis of CD20++ CD59++ ARH-77 MM cells. In addition, we demonstrate the presence of CD59 and rituximab binding on viable tumor cells from patients with MM and Waldenstrom's macroglobulinemia with progressive disease despite rituximab therapy. Last, we also examined MM and NHL B-cell lines, as well as patient tumor cells, for the expression of other antigens that may have a role in blocking ADCC activity, such as Fas ligand (FasL), MUC1, or TRAIL. FasL, MUC1, and/or TRAIL were coexpressed with complement regulators on many of these cells. These studies therefore show that complement regulators, particularly CD59 and antigens that may block ADCC, are present on various B-cell tumors and associated with rituximab resistance in patients. A prospective, clinical study is assessing the role of these antigens in mediating rituximab resistance. PMID- 11395644 TI - CD20-Directed Antibody-Mediated Immunotherapy Induces Responses and Facilitates Hematologic Recovery in Patients With Waldenstrom's Macroglobulinemia. AB - SUMMARY: Waldenstrom's macroglobulinemia (WM, lymphoplasmacytic lymphoma) is a B cell lymphoproliferative disorder in which CD20 is expressed on tumor cells from most patients. Several small studies have suggested a benefit from the anti-CD20 monoclonal antibody rituximab (Rituxan, MabThera) in patients with WM. In this retrospective study, we examined the outcome of 30 previously unreported patients with WM who received treatment with single-agent rituximab (median age 60; range 32-83 years old). The median number of prior treatments for these patients was 1 (range 0-6), and 14 patients (47%) received a nucleoside analogue before rituximab therapy. Patients received a median of 4.0 (1-11.3) infusions of rituximab (375 mg/m2). Three patients received steroids with their infusions for prophylaxis of rituximab-related infusion syndrome. Overall, treatment was well tolerated. Median immunoglobulin M (IgM) levels for all patients declined from 2,403 mg/dL (range 720-7639 mg/dL) to 1,525 mg/dL (range 177-5,063 mg/dL) after rituximab therapy (p = 0.001), with 8 of 30 (27%) and 18 of 30 (60%) patients demonstrating >50% and >25% decline in IgM, respectively. Median bone marrow lymphoplasmacytic (BM LPC) cell involvement declined from 60% (range 5-90%) to 15% (range 0-80%) for 17 patients for whom pre-and post-BM biopsies were performed (p < 0.001). Moreover, 19 of 30 (63%) and 15 of 30 (50%) patients had an increase in their hematocrit (HCT) and platelet (PLT) counts, respectively. Before rituximab therapy, 7 of 30 (23.3%) patients were either transfusion or erythropoietin dependent, whereas only 1/30 (3.3%) patients required transfusions (no erythropoietin) after rituximab. Overall responses after treatment with rituximab were as follows: 8 (27%) and 10 (33%) of the patients achieved a partial (PR) and a minor (MR) response, respectively, and an additional 9 (30%) of patients demonstrated stable disease (SD). No patients attained a complete response. The median time to treatment failure for responding (PR and MR) patients was 8.0 months (mean 8.4; range 3-20+ months), and 5.0 months (mean 6.1; range 3-12+ months) for patients with SD. These studies therefore demonstrate that rituximab is an active agent in WM. Marked increases in HCT and PLT counts were noted for most patients, including patients with WM who had MR or SD. A prospective clinical trial to more completely define the benefit of single-agent rituximab in patients with WM has been initiated by many of our centers. PMID- 11395645 TI - [Chronic arsenicism]. PMID- 11395646 TI - [Herpes zoster: incidence study among "sentinel" general practitioners]. AB - INTRODUCTION: Herpes zoster is a frequent disease but its incidence in France is unknown. METHODS: We conducted a postal survey among the general practitioners of the "Sentinel" network. The incidence of acute herpes zoster was extrapolated from the number of cases diagnosed during the year 1998, by the general practitioners who answered the questionnaire. The general practitioners were also surveyed on their prescriptions and attitude. RESULTS: Among the 1,368 "Sentinel" general practitioners, 744 (54.4 p. 100) participated in the survey. The incidence in 1998 was 3.2 cases for 1,000 inhabitants (95 p. 100 confidence interval: 3.0 - 3.4). For acute herpes zoster, 73 p. 100 of the patients have been given an oral antiviral drug, and 63 p. 100 an antalgic. Among the 605 reported herpes zoster cases, 111 (18.4 p. 100) subsequently had chronic pain. DISCUSSION: The estimated incidence is comparable to the incidence from others developed countries. To be interpreted, this estimation has to be discussed according to the sample of population that was studied and the representativity of the "Sentinelles" general practitioners who participated the survey. PMID- 11395647 TI - [Melanoma in xeroderma pigmentosum: 12 cases]. AB - BACKGROUND: Xeroderma pigmentosum is a rare genodermatosis, with a defect affecting recovery of ultraviolet-induced damages and characterized by a high rate of malignancies of the exposed skin areas. We studied melanoma features of patients with xeroderma pigmentosum. PATIENTS AND METHODS: A retrospective study of xeroderma pigmentosum patients admitted to the Charles Nicolle Hospital of Tunis between 1973 and 1998. RESULTS: Two hundred sixteen patients with xeroderma pigmentosum were registered. Melanoma was present in 12 patients, 7 females and 5 males. Two patients were sisters. Cutaneous melanoma was found in 8 patients. Four patients presented with metastatic melanoma. The median age for development of the first melanoma was 17.5 years. All of the cutaneous melanomas were found on the face. Lentigo malignant melanoma was reported in 3 cases. The tumors were treated with surgical excision. Except for a melanoma affecting the orbit, characterized by a fatal outcome, no metastases were detected at the different investigations. DISCUSSION: Melanoma occurs frequently in patients with xeroderma pigmentosum, it has been reported in 5.5 p. 100 of cases and 11.3 p. 100 of patients with cutaneous carcinoma. The age of onset was low: 17.5 years. It appeared later than the carcinoma. The location of cutaneous melanoma in face in xeroderma pigmentosum patients indicates that they were caused mainly by sunlight exposure. Lentigo malignant melanoma was the most frequent type. Prognosis is difficult to define owing to the large number of other cutaneous malignancies. Apart from one case of rapidly fatal orbital melanoma, we recorded long survivals even in cases of melanoma revealed by metastases. PMID- 11395648 TI - [Cutaneous adverse drug reactions: enhanced imputation score by skin testing]. AB - INTRODUCTION: A careful diagnosis and the identification of the causative drug after a cutaneous adverse reaction can avoid relapses. Skin tests facilitate the identification of the causative molecule by producing a hypersensitivity reaction at the application site. Adverse drug reactions are reported to Pharmacovigilance Centres who determine the imputation score of the suspected drugs. The aim of this study was to assess to what extent skin testing after a suspected allergic drug reaction can be helpful to identify the causative drug and whether an impact on the final imputation score could be evidenced. METHOD: A 18-month prospective study was performed. All patients with a history of cutaneous adverse drug reaction of suspected immunoallergic origin were included provided skin tests could be performed within 6 to 12 weeks after the adverse drug reaction. The imputation score was determined using the French imputation method prior to and after skin testing. RESULTS: Thirthy-nine patients were included in the study. Positive skin tests were observed in 11 of 20 patients with a C2S2 (I2: plausible) initial imputation score and in 6 of 15 patients with a C2S1 (I1: doubtful) initial imputation score. One patient with a C1S1 (I1: doubtful) initial imputation score had positive skin tests. DISCUSSION: The results of skin tests helped change the imputation score of the suspected drug in 18 patients out of 39. In 55 p. 100 of cases, the imputation score was increased from C2S2 (I2) to C2S3 (I3: likely) and from C2S1 (I1) to C2S3 (I3) in 40 p. 100 of cases. The increase in imputation scores was helpful to improve warning signals after an immunoallergic reaction. Skin tests led to a more accurate diagnosis in 50 p. 100 of cases. Thus, more adequate advices for further drug treatment were possible, particularly in avoiding the irrelevant prohibition of innocent drugs. PMID- 11395649 TI - [Dermatological consultation behind bars: an analysis on a three-year period in a French prison]. AB - BACKGROUND: Since 1994, health care in French prisons is managed by the public hospital system. The hospitals created ambulatory consultation units for prisons and detention centers. Skin problems rapidly became a frequent reason for consultation. This is the first systematic assessment of needs concerning dermatological consultation in prisons. MATERIAL AND METHODS: A monthly dermatology consultation was established at the Colmar detention center in May 1996. We reviewed the files of all prisoners who attended between May 1996 and May 1999. Data collected concerned motivation for the consultation, drug use and serology for HIV and hepatitic C virus. RESULTS: There were 280 consultations for 180 prisoners. All were men, mean age 30 years, who represented 16% of the total prisoner population. Most consulted for one or more benign skin conditions. The two most frequent conditions were acne (29%) and mycoses (13%). Drug use was found in 36% of the consulting prisoners (64/180). One was HIV-positive. Hepatitis C virus serology was positive in 19% of those tested (28/149), and in 39% of the drug users and 2% of the non-drug users. Motivation for consultation did not differ between drug users and non users, neither between seropositive and seronegative patients for hepatitis C virus. DISCUSSION: This survey confirms the demand for dermatology consultation in the prison population and points out the frequency of hepatitis C virus in prisoners. A monthly specialized consultation was able to meet the basic demands concerning dermatology. Adapted management of hepatitis C virus in the prison population setting remains a difficult challenge. PMID- 11395650 TI - [Dacarbazine]. PMID- 11395651 TI - [Chronic arsenicism]. AB - BACKGROUND: Arsenic is an ubiquitous natural element. Chronic and low level ingestion or inhalation may result in chronic arsenicism first characterized by skin changes. CASE REPORT: A 75 year old man, non-insulin-dependent diabetic, presented a diffuse hyperpigmentation with scattered white spots on the trunk. He complained of asthenia. Clinical diagnosis of chronic arsenicism was confirmed by arsenic determination in urine, plasma and phaneres. Thorough investigations led to discover very high arsenic levels in the own wine of the patient. This was probably the result of a wrong use of sodium arsenite-based fungicide, for cultivating his vine yard. DISCUSSION: Chronic arsenicism has become rare but it should always be kept in mind. Clinical presentation, with particular cutaneous features and routes of exposure are reviewed. Treatment is symptomatic. As arsenic is known to be a strong carcinogenic agent, patients with chronic arsenicism have to be followed up during a long time. PMID- 11395652 TI - [Dyshidrosis and acral purpura during polymorphic dermatitis in pregnancy: 2 cases]. AB - BACKGROUND: We describe two severe cases of polymorphic eruption of pregnancy associated with dyshidrosis and purpuric lesions. CASE-REPORT: Two patients, primigravida at 35 and 34 weeks of amenorrhea, were admitted in our department for a severe and itchy eruption. In the first case, some papular lesions were particularly concentrated on the abdominal striae, associated with vesicles on the limbs. There were plantar dyshidrosis and purpuric macules on the toes. In the second case, there were a maculopapular eruption on the abdomen and limbs, some vesicles and bullae of palms and soles, and a purpura of heels. In both cases, the skin biopsy showed dermic inflammatory, direct immunofluorescence and Pemphigoide Gestationis Factor research were negatives. The two patients were treated with betamethasone dipropionate (Diprosone(R)), and hydroxyzine (Atarax(R)). They gave birth to healthy babies. DISCUSSION: Dyshidrosis is not exceptional in polymorphic eruption of pregnancy. Indeed, in the several series of polymorphic eruption of pregnancy we checked, a pompholyx distribution was observed in 5 p. 100 of the cases. It is not necessary a sign of pemphigoid gestationis, even if this localization is commonly observed. The purpuric lesions are unusual because, according to our knowledge, there is no description of purpura associated with palmoplantar lesions in polymorphic eruption of pregnancy. The purpuric lesions may be secondary to high capillary permeability, increased by severe dermic inflammation of the polymorphic eruption of pregnancy and, possibly, high level of circulating estrogens. PMID- 11395653 TI - [Subcutaneous inflammatory edema induced by MINE chemotherapy]. AB - BACKGROUND: The MINE regimen (mitoguazone, ifosfamide, vinorelbine and etoposide) is a salvage chemotherapy for relapsed and refractory Hodgkin's disease. CASE REPORTS: We report the cases of a 16-year-old girl and a 17-year-old boy who had Hodgkin's disease and developed painful and massive subcutaneous inflammatory edema after MINE chemotherapy. Morphine was unable to control pain leading to major functional disability of joint movement. One patient had an elevated creatine kinase level, hypoalbuminemia, hypodermic and muscular edema at magnetic resonance imaging and diffuse hemorrhagic hypodermic edema at skin biopsy. The other patient was found to have only hypoalbuminemia. The clinical course was favorable in both cases within a few weeks, but with recurrent episodes of pain and localized areas of fat necrosis five months later in one case. DISCUSSION: This side effect of MINE chemotherapy - subcutaneous inflammatory edema, myalgia and skin pain - has not been described previously for the different components of the regimen. Three clinicopathological hypotheses could be put forward: capillary leak syndrome, panniculitis, toxic fasciitis. The causal drug remains undetermined, but the most likely would be vinorelbine because of the chronology of the eruptions during the first and last days of chemotherapy and because of the known vascular toxicity of vinorelbine which could explain a capillary leak syndrome. PMID- 11395654 TI - [Olmsted syndrome]. AB - BACKGROUND: The Olmsted syndrome is a rare congenital disorder with palmoplantar keratoderma and hyperkeratotic periorifical plaques. Only 19 cases were reported in the literature. Other cutaneous and organic findings have been reported with different frequencies. CASE REPORT: A 17-year-old girl presented with genital, perigenital and perianal hyperkeratotic plaques extending to the thighs, palmoplantar keratoderma and nail dystrophy. A small erythematokeratotic plaque was localized below the lower lip. Improvement of the palmoplantar and genital lesions was noted after one month of treatment with 25 mg acitretine daily. Further substantial improvement of the palmoplantar and ungueal lesions was observed during the next 7 months. However, discrete erythematous perianal and perigenital lesions persisted in these areas 2 years later. DISCUSSION: Efficacy of retinoids in the Olmsted syndrome is controversially discussed in the literature. However, acitretin seems to be the treatment of choice of this syndrome in adults. PMID- 11395655 TI - [Granuloma annulare following BCG vaccination]. AB - BACKGROUND: Granuloma annulare following BCG vaccination or tuberculin skin tests is rarely reported in the literature. We describe three cases occurring in patients under the age of 6 years. CASE REPORTS: In the first two cases, granuloma annulare was initially localized at the vaccinal site, then generalized. In the third case, diagnosis was deep granuloma annulare localized far from the initial vaccination site, with recurrence following tuberculin test. In the three cases, diagnosis was made on the basis of clinical and histological elements. DISCUSSION: The relationship between granuloma annulare and BCG may seem evident when the eruption occurs at the vaccinal site, but remains difficult to prove. Diagnosis essentially relies on the absence of other classical etiologies and a compatible delay. When a vaccination such as BCG appears to be the cause of granuloma annulare, a possible cause would be injection trauma or a cell-mediated-delayed-hypersensitivity reaction to a specific antigen contained in the vaccine, leading to development of skin disorders such as granuloma annulare in predisposed subjects. Reports of granuloma annulare following BCG are uncommon but the incidence may be underestimated. In practice, diagnosis of granuloma annulare following BCG or tuberculin skin test cannot be established until BCG dissemination infection is ruled out, a necessary clinical and paraclinical step required in all cases of eruptions following this vaccination. Granuloma annulare is a benign complication requiring no treatment and does not constitute a contraindication for other usual vaccinations. PMID- 11395656 TI - [Eosinophilic arteritis of the scalp]. AB - BACKGROUND: The occurrence of recurrent peripheral arterial thromboses together with a blood eosinophilia generally suggests an occlusive vascular disease or a systemic vasculitis. CASE REPORT: In a 31-year-old woman with a 15-year history of severe smoking and a blood eosinophilia from 1,200 to 2,500/mm(3), we observed recurrent attacks of pruritus and urticaria and recurrent lesions of eosinophilic thromboangiitis of hypodermal medium-sized elastic arteries of the scalp. In spite of the persistent eosinophilia, the evolution was spontaneously regressive and no other clinical or biological sign occurred within a follow-up time of 3 years. DISCUSSION: In Buerger's disease (thromboangiitis obliterans) and in most systemic vasculitis, especially in Churg-Strauss syndrome, the first lesions may be inflammatory thromboses of the extra-cranial scalp arteries. The diagnosis of an eosinophilic arteritis of the scalp may only be considered if the examination of the other peripheral vessels is normal and if the course of the disease is benign, without any treatment, in spite of a persistent blood eosinophilia. This clinico-pathological presentation should be considered as a distinct entity. PMID- 11395657 TI - [Keratosic knee nodule]. PMID- 11395658 TI - [Poikilodermis]. PMID- 11395659 TI - [Management of Raynaud's phenomenon]. PMID- 11395660 TI - [Chilblain]. PMID- 11395661 TI - [Developments on paraneoplasic pemphigus]. PMID- 11395662 TI - [Neurofibromatosis 1: recommendations for management]. PMID- 11395663 TI - [Hypersensitivity to latex or natural rubber]. PMID- 11395664 TI - [Folliculitis and renewed immunocompetence in HIV-positive subjects after antiretroviral treatment]. PMID- 11395665 TI - [Skin eruptions due to azithromycin (Azadose-Zithromax) and infectious mononucleosis]. PMID- 11395666 TI - [Copahic erythema]. PMID- 11395667 TI - [Digestive toxicity of low-dose aspirin: an underestimated and unresolved problem]. PMID- 11395668 TI - [Upper gastrointestinal bleeding in patients treated by low-dose aspirin]. AB - AIM OF THE STUDY: To estimate the number of people treated by low-dose aspirin (<330 mg daily) in France and to evaluate the risk of upper gastrointestinal bleeding associated with low-dose aspirin treatment. SUBJECTS AND METHODS: One thousand six hundred and two patients with upper gastrointestinal bleeding were included between January and June 1996 in 4 French areas. Data about patients characteristics, drugs recently used, and bleeding lesions were prospectively collected. Five hundred seventy five cases were matched for sex, age and area with control people without previous upper gastrointestinal bleeding. Low-dose aspirin intake in the population was estimated from the control group. Aspirin intake in the previous 7 days in cases and in controls was compared by logistic regression, adjusted for other gastrotoxic drugs intake. RESULTS: Low-dose aspirin is taken by about 1.2 millions adults in France. In 1 602 patients, gastrointestinal bleeding was related to a peptic ulcer in 34%. Aspirin was associated with higher risk of upper gastrointestinal bleeding: OR=1.68 (1.03 2.74) with low-dose, and OR 1.42 (0.91-2.21) with higher doses. CONCLUSION: About 2.8% of the population is exposed to low-dose aspirin in France. This treatment seems to be associated with a high risk of upper gastrointestinal bleeding. PMID- 11395669 TI - [Protective effect of appendectomy on the development of ulcerative colitis. A case-control study]. AB - AIMS: To examine by a case-control study the relationship between appendectomy and subsequent ulcerative colitis development in a French population. METHODS: A total of 150 patients with ulcerative colitis were matched for age (+/- 5 years) and sex, with 150 controls recruited in a preventive medicine center. The following data were collected from medical records and by standardised questionnaire in consultation or by phone: appendectomy and tonsillectomy before the onset of ulcerative colitis, smoking habits and area of residence. RESULTS: The rate of previous appendectomy in patients with ulcerative colitis was 8% (12/150) compared with 30.6% (46/150) in the control group (P=0.001). There was no significant association between ulcerative colitis and tonsillectomy (25.3 and 27.3% in the control and the ulcerative colitis groups, respectively). Smoking was more frequent in the control group (36%) than in the ulcerative colitis group (25.3%) but the difference was not significant (P=0.07). In multivariate analysis, the risk of developing ulcerative colitis was significantly lower after previous appendectomy (odds ratio=0.26; 95% confidence interval: 0.13-0.55; P=7 x 10(-4)). CONCLUSION: Our study confirms the inverse association between appendectomy and subsequent ulcerative colitis, in a French population, after adjusting on smoking. PMID- 11395670 TI - Pantoprazole versus lansoprazole in French patients with reflux esophagitis. AB - OBJECTIVES: The aim of this study was to compare the efficacy of pantoprazole 40 mg and lansoprazole 30 mg given for 4 to 8 weeks on endoscopic healing and symptom relief in grade II-III reflux esophagitis patients (according to Savary Miller classification). METHODS: Four hundred and sixty one patients were included (pantoprazole n=226, lansoprazole n=235) in this prospective, randomized, multicenter double-blind study. Endoscopic control was performed at 4 weeks and at 8 weeks if esophagitis was not healed. RESULTS: In the intention-to treat analysis, the healing rates at 4 weeks were 81 and 80% in the pantoprazole and lansoprazole groups, respectively (NS), 90 and 86% at 8 weeks (NS). In the per-protocol analysis, the healing rates at 4 weeks were 86% in the 2 groups and at 8 weeks 97% in the pantoprazole group and 93% in the lansoprazole group (NS). The heartburn relief rates at day 14 were 88% and 86% in the pantoprazole and lansoprazole groups, respectively. Only esophagitis grade at entry was shown to be a predictive factor for healing at 4 weeks (P<0.0001). CONCLUSION: This study showed that pantoprazole 40 mg once daily is as effective and well tolerated as lansoprazole 30 mg once daily in the treatment of grade II-III acute reflux esophagitis. PMID- 11395671 TI - [Chronic intestinal pseudo-obstruction secondary to connective tissue diseases]. PMID- 11395672 TI - [Tele-surgery: mythe or reality?]. PMID- 11395673 TI - [Medical follow-up of patients with positive serology for hepatitis C virus]. AB - OBJECTIVE: A screening campaign requested by the French Health Insurance Fund was performed in the Lyon area to access the prevalence of anti-hepatitis C virus in patients from 271 general practitioners and to observe the follow-up of the patients with had positive serology. A total of 101 patients had already had hepatitis C virus antibodies and 30 patients had newly detected antibodies against hepatitis C virus. All subject received appropriate medical care. The aim of this study was to analyse medical follow-up. METHODS: Follow-up was performed for one year by the patient's general practitioner or a specialist of liver disease. Clinical, biological, histological and virological data were collected and treatment was proposed when appropriate. Information was gathered anonymously by phone. RESULTS: Follow-up was regular for 28 patients (93%). During this follow-up, ALAT levels remained within the normal limit for 13 patients (43%) and 6 of them had undetectable viral RNA by PCR. In the other patients (50%), a liver biopsy was affered to 11 patients (40%), and performed in 9 (30%). Treatment was started in 8 patients. CONCLUSION: The results of this study on medical follow-up are satisfying. The general practitioner plays a key role in the follow-up of patients infected by hepatitis C virus. PMID- 11395675 TI - [Gene therapy for hepatocellular carcinoma. From the idea to clinical trials]. PMID- 11395674 TI - [Biological factors influencing response to diuretics in patients with cirrhosis and ascites]. AB - PURPOSES: To examine the biological factors influencing response to diuretics in patients with cirrhosis and ascites. METHODS: Sixty-nine patients were evaluated. Patients were classified into 3 groups: group 1: "good responders" (responding to spironolactone 200 mg/day), group 2: "bad responders" (responding to spironolactone doses above 200 mg/day or requiring addition of furosemide), and group 3: "non-responders" (not responding to spironolactone 400 mg/day and furosemide 160 mg/day). RESULTS: There were 30 patients in group 1, 24 in group 2 and 15 in group 3. The degree of activation of the renin-aldosterone and sympathetic system in group 2 was significantly higher than in group 1 and lower than in group 3. Natriuresis in group 2 (11 +/- 0.7 mEq/24h) was significantly below group 1 (20 +/- 2 mEq/24h) and above group 3 (5 +/- 0.6 mEq/24h). CONCLUSIONS: In patients with cirrhosis and ascites, the degree of activation of the renin-aldosterone and sympathetic nervous system influences diuretic response of ascites and is estimated by measured baseline natriuresis. PMID- 11395676 TI - [Treatments for hepatitis virus C infections]. PMID- 11395677 TI - [Atrophic polychondritis associated with ulcerative colitis]. AB - We report the case of a 46-year-old patient in whom ulcerative colitis had been diagnosed three years ago. He was admitted to the hospital for swelling of the nose. Clinical course and complementary exams led us to diagnose atrophic polychondritis. Twelve cases of such an association have been published so far. PMID- 11395678 TI - [Achalasia mimicking psychiatric eating disorders]. AB - Anorexia nervosa and psychogen vomiting are psychiatric eating disorders characterized by unexplained weight loss and induced vomiting. These diagnoses require absence of somatic disease. Achalasia is a primary disorder of the esophagus that can be responsible for the same symptoms. This may occult the real diagnosis, especially as dysphagia is not constant and variable in time. We report four cases of achalasia mistakenly diagnosed and treated as anorexia nervosa or psychogen vomiting. Achalasia was unrecognized because specific symptoms, such as dysphagia, were overlooked or misinterpreted by the patients' physicians and psychiatrists, or by the patients themselves. In patients with such eating disorders considered to be psychiatric, physicians should inquire about signs suggestive of achalasia. The diagnosis of achalasia is suspected by imaging and endoscopy, and confirmed or ruled out by manometry. PMID- 11395679 TI - [Liver transplantation and constrictive pericarditis]. AB - We report the case of a patient with refractory ascitis due to a constrictive pericarditis who underwent a liver transplantation with the initial diagnosis of cryptogenic cirrhosis. The cardiac origin was suspected 5 months post surgery when a liver biopsy showed lesions in favor of a post sinusoidal shunt. The diagnosis was confirmed by the increased values of the right intra-ventricular pressures. We discuss the causes of the delay of the diagnosis and, in particular, the difficulty to interpret vascular liver lesions. Such vascular lesions were present on the needle biopsy performed prior to transplantation but wrongly interpreted as cirrhosis. PMID- 11395680 TI - [Suppurated hepatic metastasis revealing a colonic stromal tumor]. PMID- 11395682 TI - [Post-polypectomy colonic arterial hemorrhage can be treated by hemoclipping]. PMID- 11395681 TI - [Ovarian metastasis of esophagus squamous cell carcinoma]. PMID- 11395683 TI - [Systemic sclerosis and ulcerative colitis]. PMID- 11395684 TI - [Encephalopathy induced by ingestion of ammonium during valproate therapy]. PMID- 11395685 TI - [Transient portal vein thrombosis during pregnancy in a patient with cirrhosis]. PMID- 11395686 TI - [Acute hepatitis following intravenous buprenorphine injection as a substitute drug in a drug-addict]. PMID- 11395687 TI - [Evaluation of oocyte maturity in patients undergoing ICSI and R-FSH treatment]. AB - BACKGROUND: This study compares embryo quality, fertilisation rate and pregnancy rates after ICSI related with the quality of oocytes achieved after r-FSH stimulation. METHODS: We evaluated 955 oocytes from patients following r-FSH and 643 oocytes from patients stimulated with ultrapure urinary FSH. The oocytes were divided into: a) normal oocytes; b) ooctyes with extra cytoplasmatic abnormalities (dark zona pellucida, wide perivitelline space); c) oocytes with cytoplasmatic abnormalities (dark cytoplasm, granular cytoplasm, retractile body), d) oocytes with shape abnormalities; e) oocytes with double abnormalities; f) oocytes with triple abnormalities. The embryos were divided into: A) even and homogeneous blastomeres without fragmentation; B) even and homogeneous blastomeres with <20% fragmentation; C) uneven and non-homogeneous blastomeres with 20-50% fragmentation; D) uneven and non homogeneous blastomere with >50% fragmentation. RESULTS: 40.9% of oocytes from patients treated with r-FSH have a normal morphology vs 35.2% of control groups (p<0.04). Abnormalities have a similar frequency in the two groups, except for the presence of three combined abnormalities (7.7 vs 5.4%; p<0.04). Fertilisation rate, cleavage rate, oocyte quality and pregnancy rate do not appear to be influenced by oocyte morphology and the type of FSH used for stimulation. CONCLUSIONS: The administration of r FSH allows a large number of oocytes to be rescued, with a high incidence of normal morphology. The fertilisation rate and the quality of embryos obtained from oocytes with structural abnormalities are similar to those observed in morphologically normal oocytes. Even the probability of pregnancy is similar in the two groups. PMID- 11395688 TI - [Post-partum vesicovaginal fistula: abdominal muscle strip treatment]. AB - BACKGROUND: The Vesico-Vaginal Fistula (VVF) very often occur in tropical countries, but their treatment is usually not correct. METHODS: A surgical treatment was carried put on 68 patients affected by VVF from March 1986 to December 19997 in the Nazareth Hospital (Nairobi). Their mean age was 22 years old; 27 patients (39.7%) underwent surgery for the first time, while for 41 patients (60.2%) the treatment was repeated. Fourteen patients (20.5%) had also Vesico-Rectum-Vaginal Fistula (VRV). The VVF was cured with a transvaginal treatment using a Martius strip for 32 cases. For 16 cases both vaginal and abdominal treatment was performed in the same time using an abdominal muscle strip, which was inserted in the space between the vagina and bladder. In VRV and VVF combined cases, the VVF was treated in the following way: first of all, during the same session, the VVF was cured by making a colostomy and then, after 2 months VRV was treated. RESULTS: The follow-up took about 7.2 months; 62 patients (91.1%) recovered, for 6 cases treated only with transvaginal operation, it has been necessary a second surgical procedure owing to relapsing, and for 2 of them an abdominal muscle strip was used. CONCLUSIONS: In conclusion, while the transvaginal repair is satisfactory treatment for little fistula never surgically treated before, on the other hand the transabdominal vaginal treatment is the best cure forge large or relapsing fistulas. PMID- 11395689 TI - [Bleedings during the third term: which complications?]. AB - BACKGROUND: The aim of this research was to study the maternal and prenatal complications which may occur in the abruptio placentae and in placenta praevia. METHODS: All the patients who were hospitalized in the Obstetrics and Gynaecology Department of the Policlinico Umberto I, from January 1993 to July 2000 have been studied. Moreover, a comparative study between the onset of complications of the two pathologies considered and the ones found in a control group was carried out. We found 62 cases of placenta previa and 48 cases of abruptio placentae. The total amount of childbirths was 6861, so the incidence of abruptio placentae was 0.007%, while the incidence of placenta previa was 0.009%. RESULTS: The results show that the most frequent maternal complication, in both pathologies, was haemorrhage. We did not find any statistically meaningful difference between the percentage of haemorrhage in the two pathologies ( p=0.0608), but we noted a higher percentage of haemorrhage compared to the control group. The number of hysterectomies was higher in patients with placenta previa compared to patients with abruptio placentae. The most frequent fetal complication was premature birth, significantly more frequent than in the control population (p=0.0210). CONCLUSIONS: As a matter of fact, we can affirm that, in all its clinical manifestations abruptio placentae is a more dangerous disease than the placenta previa, except in the complications of the discharge of the afterbirth and of the postpartum, where haemorrhage and uterus atony lead to a great number of hysterectomies. PMID- 11395690 TI - [Vertical trasmission of human immunodeficiency virus (HIV) and other sexually transmitted infections (STI)]. AB - Infectious agents which are sexually transmitted determine considerable morbidity in women during the gestational period. Connatal and perinatal infection of the newborn, miscarriage, and low birthweight have all been described. Vertical transmission of HIV and other STD may occur via the placenta during gestation (the major mechanism for syphilis) or at birth during the passage through the cervico-vaginal channel (the major mechanism for HIV, HBV, HSV, gonorrhoea and chlamydia). High serum viral loads of HIV significantly increase the likelihood of newborn infection, while the presence of lesions in the genital tract at birth increases the odd for transmission for HSV. Breast feeding is a well described route of transmission for HIV infection, but it is irrelevant to the transmission of HBV. Cutaneous lesions of the breast and nipples carry a risk of transmission of syphilis and HSV through breast-feeding. Treatment of the etiologic agent is considered an effective means for the prevention of vertical transmission and is recommended for all STI agents except for HBV. HIV infected women on antiretroviral therapy should continue the same treatment regimen if they become pregnant (with the exception of indinavir and efavirenz, which should be replaced as soon as possible); women who did not assume antiretroviral drugs at the time they became pregnant, should start treatment as soon as they reach the second trimester of gestation. Delivery should be performed by elective cesarian section in all HIV infected women. Delivery should also be performed by cesarian section in women who develop a primary HSV infection and have cervico-vaginal lesions. Recurrent episodes of genital herpes are associated to a much lower risk of vertical transmission and do not represent a criterium for cesarian section. Women with documented cervical chlamydia infection should receive a full treatment regimen at the 36th week of gestation. Women with chronic HBV infection do not require etiologic treatment; however, their newborns should receive concomitant doses of HBV immunoglobulins and HBV vaccine soon after birth. Standard practices of prevention of vertical transmission of STI agents applies to women regardless their native country. However, the feasibility of implementation of the guidelines in poor resource countries is a matter of great concern: an unresolved debate is ongoing on optimal strategies for the prevention of vertical transmission of HIV in such countries. PMID- 11395691 TI - [Changes of leptin levels in menopause]. AB - One of the main advances in the field of metabolic control of body weight and obesity treatment was the identification of the OB protein or leptin, that plays an important role in controlling body weight, signalling to the CNS the amount of body fat. Indeed, leptin levels are positively correlated to indices of body fat, namely total fat mass, percent body fat and body mass index (BMI). This protein may be also the signal that indicates the nutritional status to the reproductive axis. Whether this signal is exerted directly on the gonads or through the neuroendocrine axis is still to be determined. A sexual dimorphism between male and female in serum leptin levels has been observed, with the latter showing higher serum leptin levels. This evidence has led to the hypothesis that estrogens might have a stimulatory role in leptin secretion. To evaluate this hypothesis, several authors have determined serum leptin levels in postmenopausal women that have estrogen levels comparable to those present in men. The results of these studies are contradictory and the aim of this article has been the revision of data present in the literature regarding serum leptin levels in menopause and to correlate them to body composition changes taking place during menopause. PMID- 11395692 TI - [Ectopic pregnancy after in vitro fertilization and embryo transfer. Analysis of literature]. AB - The incidence of ectopic pregnancy has dramatically increased in the last decade. The triggering factors may be different and among them there is the introduction of medical assisted procreation techniques. Tubal pathologies in 80% of women undergoing artificial insemination techniques, the transfer in the womb of more than one embryo and the method of the same embryo-transfer may influence the onset of an ectopic pregnancy. This study aims to detect the factors that can prevent or reduce thc incidence of this pathology, after a close examination of the literature. PMID- 11395693 TI - [Pregnancy in women with thalassaemia]. AB - Women with transfusion dependent thalassaemia suffer from failure of pubertal growth and delayed onset of menarche with amenorrhea, anovulation and infertility. With improved pediatric and hematological care is now possible, for patients with b thalassaemia, to achieve a pregnancy. Pre-pregnancy assessment included checks for hypothyroidism and diabetes, for hepatitis B and C, human immunodeficiency virus, Rubella, cardiac functions, liver functions by estimating aspartate and alanine aminotransferases, gamma-glutamyl transpeptidase, alkaline phospatase, and total plasma proteins. The frequency of blood transfusion needed to be increased in order to maintain the hemoglobin concentration above 10 g/dl. Desferroxamine must be stopped as soon as pregnancy is diagnosed continuing the administration of the folic acid supplements throughout pregnancy. Desferroxamine will be resumed after delivery. The safety of iron chelation with desferroxamine during the periconceptional period and pregnancy has not yet been established. Some animal studies have shown skeletal anomalies; other published studies report seven women with b thalassaemia major who became pregnant while taking desferroxamine: all the women had normal babies. The mode of delivery is usually vaginal, while Cesarean section is performed in those cases with pre-eclampsia, fetal distress, cephalopelvic dysproportion, slow progression of labor, as in women without thalassaemia. In conclusion, with the advent of regular blood transfusion associated with iron chelation therapy, pregnancy in b thalassaemia can be safe for mothers and their babies with appropriate care. PMID- 11395694 TI - [The EX-utero Intrapartum Technique (EXIT) procedure in Italy]. AB - Aim of the study was to present the first two Italian cases of C-section performed with the EXIT procedure (EX-utero Intrapartum Technique). Deliveries were performed at the Division of Obstetrics and Gynecology of the Hospital of Padua in cooperation with the Pediatric Surgery Department, both tertiary care centers. The first case was a twin with a huge neck mass (cystic hygroma) and the second a fetus with an oropharyngeal mass (epignathus). Airway patency could have been compromised at birth in both of them. EXIT procedure consists in securing the airway of the fetus partially delivered and still connected with the placenta. This technique leaves an intact feto-placental circulation and guarantees a normal fetal oxygenation while fetal airway patency is secured. Both the fetuses were successfully intubated and the C-section ended up in a short period of time without maternal and fetal complications. The EXIT technique, performed for the first time in 1989 and now in many centers abroad, can be considered a safe procedure as long as a multidisciplinary approach is carried out. The EXIT procedure is indicated whenever fetal airways can be compromised at birth, that is when oropharyngeal masses, laryngeal atresia, cystic hygroma and goiter are encountered during prenatal ultrasound. PMID- 11395695 TI - [Spontaneous ectopic contralateral pregnancy with unicornuate uterus. A case report]. AB - A case is reported of spontaneous ectopic pregnancy in a non patent fallopian tube of a patient with a contralateral unicornuate uterus without rudimentary horn. A hemoperitoneum, due to the rupture of the tube, occurred with a HCG level of 48 I.U./l. A laparoscopy was therefore performed and the fallopian tube removed. PMID- 11395696 TI - [Historical observations on shoulder dystocia. 1) From the origin to Jacquemier]. AB - The result of a literature survey on shoulder dystocia from year 1609 to 1860 are presented. Operative maneuvers for the resolution of dystocia were mainly examined, as well as the development of knowledge about the pathogenetic mechanism of this serious complication. PMID- 11395697 TI - Functional therapy. PMID- 11395698 TI - Long-term online search capabilities stymied. PMID- 11395699 TI - Comparison of 2 modifications of the twin-block appliance in matched Class II samples. AB - The purpose of this study was to compare the skeletal and dental changes contributing to Class II correction with 2 modifications of the Twin-block appliance: Twin-block appliances that use a labial bow (TB1) and Twin-block appliances that incorporate high-pull headgear and torquing spurs on the maxillary central incisors (TB2). After pretreatment equivalence was established, a total of 36 consecutively treated patients with the TB1 modification were compared with 27 patients treated with the TB2 modification. Both samples were treated in the same hospital department and the same technician made all the appliances. The cephalostat, digitizing package, and statistical methods were common to both groups. The results demonstrated that the addition of headgear to the appliance resulted in effective vertical and sagittal control of the maxillary complex and thus maximized the Class II skeletal correction in the TB2 sample. Use of the torquing springs resulted in less retroclination of the maxillary incisors in the TB2 sample when compared with the TB1 sample; however, this difference did not reach the level of statistical significance. PMID- 11395700 TI - Cephalometric markers to consider in the treatment of Class II Division 1 malocclusion with the bionator. AB - The principal aims of this study were to find the cephalometric predictors for good treatment results of bionator therapy and to justify bionator therapy as an early phase I treatment for patients with Class II Division 1 malocclusion. Forty subjects who had Class II Division 1 malocclusion and who were treated with the bionator were selected for this study. They were classified into a good treatment result group and a poor treatment result group in reference to the posttreatment molar relationship, posttreatment overbite and overjet, posttreatment profile, and existence of relapse. Thirty-one cephalometric variables were analyzed on the pretreatment lateral cephalograms by t-test to evaluate differences between the 2 groups. The variables that differed significantly were analyzed by discriminant analysis to assess their predictability. The study showed that horizontal growth pattern, close to normal anteroposterior relationship between the maxilla and mandible, upright mandibular incisor, and retrusive lower lip were important predictors for good results. In particular, protrusion of the lower lip was the most important factor for the determination of the treatment results. In addition, the treatment times were much shorter for patients with these good predictors. The present study supports the premise that bionator therapy can produce clinically stable and favorable results if the patients are appropriately selected with the use of these diagnostic criteria. PMID- 11395701 TI - Pseudoelasticity and thermoelasticity of nickel-titanium alloys: a clinically oriented review. Part I: Temperature transitional ranges. AB - The purpose of this review was to organize a systematic reference to help orthodontists evaluate commonly used orthodontic nickel-titanium alloys. Part I of the article reviews the data available in the literature regarding the temperature transitional ranges of the alloys. The thermomechanical behavior of these compounds is, in fact, strictly dependent on the correlation between the temperature transitional range and the oral temperature range. Part II of the article will focus on the mechanical characteristics of the alloys, such as the magnitude of the forces delivered and its correlations with temperature transitional range and oral temperature. PMID- 11395702 TI - Pseudoelasticity and thermoelasticity of nickel-titanium alloys: a clinically oriented review. Part II: Deactivation forces. AB - The purpose of this review was to organize a systematic reference to help orthodontists evaluate commonly used orthodontic nickel-titanium alloys. Part I of the article reviewed the data available in the literature regarding the temperature transitional ranges of the alloys. The thermomechanical behavior of these compounds is, in fact, strictly dependent upon the correlation between the temperature transitional range and the oral temperature range. Part II focuses on the mechanical characteristics of the alloys, such as the magnitude of the forces delivered and correlations with the temperature transitional range and oral temperature. PMID- 11395703 TI - Estimation of force produced by nickel-titanium superelastic archwires at large deflections. AB - A method for estimating the absolute values of forces produced by nickel-titanium orthodontic wires at superelastic deformations is proposed. The method is based on both the elasticity theory and the experimental bending tests for the nickel titanium wires. Forces for a number of nickel-titanium wires are calculated by this method. PMID- 11395704 TI - Rapid curing of bonding composite with a xenon plasma arc light. AB - The use of light-cured orthodontic adhesives is an increasingly popular method for the bonding of orthodontic brackets. However, one of the disadvantages of light-cured adhesives is their long curing times. The xenon plasma arc curing light is purported to dramatically reduce the required curing time. The purpose of this study was to test the efficiency of a xenon plasma arc light versus a conventional tungsten-quartz halogen light in producing effective bond strengths for orthodontic brackets. Standardized brackets were bonded to bovine enamel with 3 different orthodontic bonding materials. The bonding materials were exposed to the tungsten-quartz halogen light for 40 seconds and to the xenon light for 3, 6, and 9 seconds. Bond strength was tested 30 minutes and 24 hours after light curing. The results showed that bond strength with the application of the xenon light was greater with longer exposures. There were no statistically significant differences between the bond strengths of the brackets exposed to the tungsten quartz halogen light for 40 seconds and those exposed to the xenon light for 3, 6, or 9 seconds. However, xenon light exposures of 6 or 9 seconds were required to create bond strengths equal to those produced by the tungsten-quartz halogen light. The xenon light produced equivalent bond strengths at very short light exposures. PMID- 11395705 TI - Tensile bond strength of ceramic orthodontic brackets bonded to porcelain surfaces. AB - The aim of this study was to compare various surface treatment methods to define the procedure that produces adequate bond strength between ceramic brackets and porcelain. The specimens used in this study, 60 porcelain tabs, were produced by duplication of the labial surface of a maxillary first premolar. The 6 different preparation procedures tested were: (1) sandblasting with 50 microm aluminum oxide in a sandblasting device, (2) application of silane to the porcelain and the bracket base, (3) sandblasting followed by application of silane, (4) acid etching with 9.6% hydrofluoric acid, (5) acid etching with 9.6% hydrofluoric acid followed by application of silane, and (6) sandblasting followed by application of 4-Meta adhesive. The ceramic brackets were bonded with no-mix orthodontic bonding material. A bonding force testing machine was used to determine tensile bond strengths at a crosshead speed of 0.5 mm per second. The results of the study showed that porcelain surface preparation with acid etching followed by silane application resulted in a statistically significant higher tensile bond strength (P < .05). Sandblasting the porcelain surface before silane treatment provided similar bond strengths, but sandblasting or acid etching alone were less effective. Silane application was recommended to bond a ceramic bracket to the porcelain surface to achieve bond strengths that are clinically acceptable. PMID- 11395706 TI - Effect of a self-etch primer/adhesive on the shear bond strength of orthodontic brackets. AB - Conventional adhesive systems use 3 different agents (an enamel conditioner, a primer solution, and an adhesive resin) during the bonding of orthodontic brackets to enamel. A unique characteristic of some new bonding systems in operative dentistry is that they combine the conditioning and priming agents into a single product. Combining conditioning and priming saves time and should be more cost-effective to the clinician and, indirectly, to the patient. The purpose of this study was to determine the effects of the use of a self-etch primer on the shear bond strength of orthodontic brackets and on the bracket/adhesive failure mode. Brackets were bonded to extracted human teeth according to 1 of 2 protocols. In the control group, teeth were etched with 37% phosphoric acid. After the sealant was applied, the brackets were bonded with Transbond XT (3M Unitek, Monrovia, Calif) and light cured for 20 seconds. In the experimental group, a self-etch acidic primer (ESPE Dental AG, Seefeld, Germany) was placed on the enamel for 15 seconds and gently evaporated with air, as suggested by the manufacturer. The brackets were then bonded with Transbond XT as in the first group. The present in vitro findings indicate that the use of a self-etch primer to bond orthodontic brackets to the enamel surface resulted in a significantly (P = .004) lower, but clinically acceptable, shear bond force (mean, 7.1 +/- 4.4 MPa) as compared with the control group (mean, 10.4 +/- 2.8 MPa). The comparison of the adhesive remnant index scores indicated that there was significantly (P = .006) more residual adhesive remaining on the teeth that were treated with the new self-etch primer than on those teeth that were bonded with the use of the conventional adhesive system. PMID- 11395707 TI - Enhancing wire-composite bond strength of bonded retainers with wire surface treatment. AB - Bonded orthodontic retainers with wires embedded in composite resin are commonly used for orthodontic retention. The purpose of this study was to test, in vitro, various wire surface treatments to determine the optimal method of enhancing the wire-composite bond strength. Coaxial wires and stainless steel wires with different surface treatments were bonded to bovine enamel and then pulled along their long axes with an Instron universal testing machine. Wire surface treatments included placing a right-angle bend in the wire, microetching the wire, and treating the wire with adhesion promoters; combinations of treatments were also examined. The results demonstrated a 24-fold increase in the wire composite bond strength of wire that was microetched (sandblasted), compared with that of untreated straight wire. The difference between the amount of force required to break the bond produced by microetching alone (246.1 +/- 46.0 MPa) and that required for the bonds produced by the retentive bend (87.8 +/- 16.3 MPa), the adhesion promoters (silane, 11.0 +/- 3.1 MPa; Metal Primer, 28.5 +/- 15.8 MPa), or for any combination of surface treatments, was statistically significant. Microetching a stainless steel wire produced a higher wire-composite bond strength than that obtained from a coaxial wire (113.5 +/- 27.5 MPa). The results of this study indicate that microetching or sandblasting a stainless steel wire significantly increases the strength of the wire-composite bond. PMID- 11395708 TI - The "Tip-Edge" torquing mechanism: a mathematical validation. AB - The Tip-Edge (TP Orthodontics, LaPorte, Ind) bracket is unique among orthodontic brackets because it can be used in combination with rectangular archwire to torque teeth, not through activation of the archwire itself but through the action of auxiliary springs that are active in the tip plane. The purpose of this report is to describe the way in which a tipping couple generates a secondary torquing couple and the resulting path traced by the root apex and to express those actions mathematically. The magnitude of forces applied by the auxiliary spring to the tooth apex is calculated on the basis of an experiment. PMID- 11395709 TI - Cephalometric evaluation of craniofacial pattern of Syrian children with Class III malocclusion. AB - The purpose of this study was to investigate the morphologic characteristics of the craniofacial complex of Syrian children with Class III malocclusion. Lateral cephalometric radiographs of 69 patients with Class III malocclusion (23 male and 46 female; ages 5 to 12 years) were selected on the basis of molar relationship. Cases were analyzed and compared with a Class I control group that was matched for age, sex, and ethnic origin. The children with Class III malocclusion exhibited a distinct craniofacial morphologic characteristic that was manifest in a combination of alterations in angular and linear measurements on the lateral cephalogram. Both the anterior cranial base (SN) and posterior cranial base (SAr) were significantly shorter than normal in the Class III group, and the cranial base angle (NSAr) was slightly smaller than normal. Maxillary length (Co-A) was significantly smaller, and the maxilla was more posteriorly positioned in the patients with Class III malocclusion. The mandible was within the neutral range of protrusion, and there was a slight increase in total mandibular length (Co Gn), accompanied by a more forward positioning of the glenoid fossa in patients with Class III malocclusion. Dental aberrations in the patients with Class III malocclusion were manifested essentially by a significant decrease in the angulation and protrusion of the maxillary incisors relative to the A-Pog line, whereas the mandibular incisors showed only a slight amount of linguoversion. Patients with Class III malocclusion also tended to have a significantly smaller vertical face dimension and shorter lower anterior facial height (ANS-Me). Because of these distinct morphologic features, early orthopedic intervention with protraction face mask therapy may be the method of choice for most of the patients with Class III malocclusion included in this study. PMID- 11395710 TI - Growth hormone receptor gene variant and mandibular height in the normal Japanese population. AB - This study was aimed at quantitatively evaluating the relationship between craniofacial morphology and the Pro561Thr (P56IT) variant in the growth hormone receptor gene (GHR), which is considered to be an important factor in craniofacial and skeletal growth. The subjects were unrelated individuals in a normal Japanese population and consisted of 50 men and 50 women. With the use of genomic DNA extracted from whole blood, the GHR gene P56IT variant was detected by the polymerase chain reaction-restriction fragment length polymorphism method (with the restriction enzyme StuI). The relationships of the genotypes to body height and 5 linear measurements from lateral cephalograms were examined for evaluation of craniofacial morphology. The normal Japanese population without P56IT had a significantly greater mandibular ramus length (condylion-gonion) than did those with P56IT. This suggests that the GHR gene P56IT variant may be associated with mandibular height growth and can be a genetic marker for it. Further studies about such genetic markers may expand our understanding of the genetic control in craniofacial morphological determinants and help in the prediction of craniofacial growth. PMID- 11395711 TI - Treatment of a Class II Division 1 malocclusion with macrodontia of the maxillary central incisors. PMID- 11395712 TI - The use of educational-psychological principles in orthodontic practice. PMID- 11395713 TI - Three-dimensional on-screen virtual models. PMID- 11395714 TI - Litigation, legislation, and ethics: subsequent remedial measures. PMID- 11395716 TI - Vascular nursing in the 21st century. PMID- 11395717 TI - Postoperative anticoagulation in vascular surgery: part 1. A retrospective comparison of clinical outcomes for unfractionated heparin versus low-molecular weight heparin. AB - The use of postoperative anticoagulation is not uncommon for patients undergoing vascular procedures, whether for adjunctive therapy to the surgical procedure or for resumption of preoperative anticoagulation. Longer postoperative length of stay is necessary to achieve an adequate therapeutic international normalized ratio with traditional protocols that call for the use of unfractionated heparin. We retrospectively examined 195 cases to determine whether low-molecular-weight heparin, specifically enoxaparin, was an effective postoperative replacement for intravenous unfractionated heparin, with the impact on postoperative length of stay. There was no difference in the frequency of complications except for increased incidence of return to surgery for graft thrombosis (n = 11) in patients who received traditional intravenous unfractionated heparin with adjusted-dose warfarin daily (n = 139, P <.02). For all 195 vascular procedures combined, the average postoperative length of stay with use of enoxoparin was shortened with use of enoxaparin (P <.0008). There was a 2-day reduction in the average postoperative length of stay for the femoral-distal procedure group (n = 18, P <.004). In the first part of the article, we summarize our findings and offer new applications and ideas for vascular nurses to consider when postoperative anticoagulation is indicated after vascular procedures. Factors contributing to safe and efficacious postoperative anticoagulation with use of low-molecular-weight heparin, specifically enoxaparin, are presented to the vascular nurse. In the second part, we discuss the roles of members of the vascular team, as well as our experience with use of enoxaparin as implemented in the clinical setting. PMID- 11395718 TI - Case study: a unique approach to compliance in a patient with venous ulcers. AB - Lower extremity venous ulcers affect 500,000 to 700,000 Americans, with a 10-week outpatient treatment cost of $1327 to $5289. Recurrence rates are reported as 57% within 10.4 months. Many types of treatments are available, but the most successful treatment continues to be the most basic-elevation and compression. Innovative ideas to increase patient compliance can be the key to successful therapy. PMID- 11395719 TI - Transcutaneous oxygen measurements: implications for nursing. AB - Transcutaneous oxygen measurement is a noninvasive diagnostic technique that records the partial pressure of oxygen at the skin surface. This measurement provides information regarding the supply and delivery of oxygen to the underlying microvascular circulation. It can be used for adults in wound evaluation, hyperbaric therapy, plastic surgery, amputation level determination, and peripheral vascular disease assessment, including the status of limb revascularization procedures. This article presents an overview of transcutaneous oxygen measurement and situations that influence measurement levels, including environmental, dermal, physiologic, and mechanical factors. Nursing considerations and indications for further research are also discussed. PMID- 11395720 TI - Evidence-based practice. PMID- 11395721 TI - What is research? PMID- 11395722 TI - Pulmonary emboli. PMID- 11395723 TI - Towards a 'knowledge nation'. PMID- 11395724 TI - Researchers unnerved by echoes of the past in Russian directive. PMID- 11395727 TI - French unions walk out of talks over 35-hour week. PMID- 11395726 TI - Patent ruling could cut PCR enzyme prices. PMID- 11395728 TI - Review urges US physicists to think globally. PMID- 11395730 TI - Rivals clash over plans to take science to London public. PMID- 11395731 TI - Museum director quits over Smithsonian restructuring. PMID- 11395733 TI - Aurora project set to pave way for human space flight. PMID- 11395732 TI - Mars rock samples condemned to quarantine. PMID- 11395735 TI - Tabletop astrophysics. PMID- 11395736 TI - Biology's name game. PMID- 11395737 TI - Misclassification of pest as 'fungus' puts vital research on wrong track. PMID- 11395738 TI - Who is prepared to pay, and how much? PMID- 11395739 TI - Drug test warning. PMID- 11395745 TI - Physics in the noise. PMID- 11395744 TI - Sulphur and holy water. PMID- 11395746 TI - Fifty years of inactivation. PMID- 11395747 TI - Astronomy. A twisted look at the X-ray sky. PMID- 11395749 TI - Cancer. Telomerase meets its mismatch. PMID- 11395751 TI - Quantum electronics. Nanotubes go ballistic. PMID- 11395752 TI - Statistical physics. Advances in aggregation. PMID- 11395755 TI - Successful invasion of a floral market. PMID- 11395756 TI - Magnetic minerals. Keystone-like crystals in cells of hornet combs. AB - The hexagonal brood-rearing cells inside the nest combs of the hornet Vespa orientalis are uniform in both their architecture and orientation. We have discovered that each cell contains a minute crystal that projects down from the centre of its domed roof and has a composition typical of the magnetic mineral ilmenite. These tiny crystals form a network that may act like a surveyor's spirit-level, helping the hornets to assess the symmetry and balance of the cells and the direction of gravity while they are building the comb. PMID- 11395757 TI - Alzheimer's disease. Molecular consequences of presenilin-1 mutation. AB - Alzheimer's disease is characterized by accumulation in the brain of a family of insoluble amyloid peptides (Abeta peptides), which are produced as a result of the normal processing of beta-amyloid precursor protein (beta-APP). Russo et al. claim that a truncated Abeta peptide that lacks the first ten amino acids accumulates in the brains of patients carrying a mutant form of pre-senilin 1 (PS1), a protein that is involved in cleavage of beta-APP. However, we have found that this same species is also overrepresented in Alzheimer's patients with mutations in beta-APP itself. Our findings do not support the conclusion of Russo et al. that pathogenic PS1 mutations may control cleavage of beta-APP by beta secretase. PMID- 11395759 TI - Nanoindentation. Simulation of defect nucleation in a crystal. AB - Nanoindentation is the penetration of a surface to nanometre depths using an indenting device. It can be simulated using the Bragg bubble-raft model, in which a close-packed array of soap bubbles corresponds to the equilibrium positions of atoms in a crystalline solid. Here we show that homogeneous defect nucleation occurs within a crystal when its surface roughness is comparable to the radius of the indenter tip, and that the depth of the nucleation site below the surface is proportional to the half-width of the contact. Our results may explain the unusually high local stress required for defect nucleation in nano-indented face centred cubic crystals. PMID- 11395760 TI - Potassium channel receptor site for the inactivation gate and quaternary amine inhibitors. AB - Many voltage-dependent K+ channels open when the membrane is depolarized and then rapidly close by a process called inactivation. Neurons use inactivating K+ channels to modulate their firing frequency. In Shaker-type K+ channels, the inactivation gate, which is responsible for the closing of the channel, is formed by the channel's cytoplasmic amino terminus. Here we show that the central cavity and inner pore of the K+ channel form the receptor site for both the inactivation gate and small-molecule inhibitors. We propose that inactivation occurs by a sequential reaction in which the gate binds initially to the cytoplasmic channel surface and then enters the pore as an extended peptide. This mechanism accounts for the functional properties of K+ channel inactivation and indicates that the cavity may be the site of action for certain drugs that alter cation channel function. PMID- 11395761 TI - An efficient photoelectric X-ray polarimeter for the study of black holes and neutron stars. AB - The study of astronomical objects using electromagnetic radiation involves four basic observational approaches: imaging, spectroscopy, photometry (accurate counting of the photons received) and polarimetry (measurement of the polarizations of the observed photons). In contrast to observations at other wavelengths, a lack of sensitivity has prevented X-ray astronomy from making use of polarimetry. Yet such a technique could provide a direct picture of the state of matter in extreme magnetic and gravitational fields, and has the potential to resolve the internal structures of compact sources that would otherwise remain inaccessible, even to X-ray interferometry. In binary pulsars, for example, we could directly 'see' the rotation of the magnetic field and determine if the emission is in the form of a 'fan' or a 'pencil' beam. Also, observation of the characteristic twisting of the polarization angle in other compact sources would reveal the presence of a black hole. Here we report the development of an instrument that makes X-ray polarimetry possible. The factor of 100 improvement in sensitivity that we have achieved will allow direct exploration of the most dramatic objects of the X-ray sky. PMID- 11395762 TI - Fabry - Perot interference in a nanotube electron waveguide. AB - The behaviour of traditional electronic devices can be understood in terms of the classical diffusive motion of electrons. As the size of a device becomes comparable to the electron coherence length, however, quantum interference between electron waves becomes increasingly important, leading to dramatic changes in device properties. This classical-to-quantum transition in device behaviour suggests the possibility for nanometer-sized electronic elements that make use of quantum coherence. Molecular electronic devices are promising candidates for realizing such device elements because the electronic motion in molecules is inherently quantum mechanical and it can be modified by well defined chemistry. Here we describe an example of a coherent molecular electronic device whose behaviour is explicitly dependent on quantum interference between propagating electron waves-a Fabry-Perot electron resonator based on individual single-walled carbon nanotubes with near-perfect ohmic contacts to electrodes. In these devices, the nanotubes act as coherent electron waveguides, with the resonant cavity formed between the two nanotube-electrode interfaces. We use a theoretical model based on the multichannel Landauer-Buttiker formalism to analyse the device characteristics and find that coupling between the two propagating modes of the nanotubes caused by electron scattering at the nanotube electrode interfaces is important. PMID- 11395763 TI - Non-Fermi-liquid behaviour in La4Ru6O19. AB - Understanding the complexities of electronic and magnetic ground states in solids is one of the main goals of solid-state physics. Transition-metal oxides have proved to be particularly fruitful in this regard, especially for those materials with the perovskite structure, where the special characteristics of transition metal-oxygen orbital hybridization determine their properties. Ruthenates have recently emerged as an important family of perovskites because of the unexpected evolution from high-temperature ferromagnetism in SrRuO3 to low-temperature superconductivity in Sr2RuO4 (refs 1, 2). Here we show that a ruthenate in a different structural family, La4Ru6O19, displays a number of highly unusual properties, most notably non-Fermi-liquid behaviour. The properties of La4Ru6O19 have no analogy among the thousands of previously characterized transition-metal oxides. Instead, they resemble those of CeCu6-xAux-a widely studied f-electron based heavy fermion intermetallic compound that is often considered as providing the best example of non-Fermi-liquid behaviour. In the ruthenate, non-Fermi liquid behaviour appears to arise from just the right balance between the interactions of localized electronic states derived from Ru-Ru bonding and delocalized states derived from Ru-O hybridization. PMID- 11395764 TI - Self-assembly of regular hollow icosahedra in salt-free catanionic solutions. AB - Self-assembled structures having a regular hollow icosahedral form (such as those observed for proteins of virus capsids) can occur as a result of biomineralization processes, but are extremely rare in mineral crystallites. Compact icosahedra made from a boron oxide have been reported, but equivalent structures made of synthetic organic components such as surfactants have not hitherto been observed. It is, however, well known that lipids, as well as mixtures of anionic and cationic single chain surfactants, can readily form bilayers that can adopt a variety of distinct geometric forms: they can fold into soft vesicles or random bilayers (the so-called sponge phase) or form ordered stacks of flat or undulating membranes. Here we show that in salt-free mixtures of anionic and cationic surfactants, such bilayers can self-assemble into hollow aggregates with a regular icosahedral shape. These aggregates are stabilized by the presence of pores located at the vertices of the icosahedra. The resulting structures have a size of about one micrometre and mass of about 1010 daltons, making them larger than any known icosahedral protein assembly or virus capsid. We expect the combination of wall rigidity and holes at vertices of these icosahedral aggregates to be of practical value for controlled drug or DNA release. PMID- 11395765 TI - Stability of atmospheric CO2 levels across the Triassic/Jurassic boundary. AB - The Triassic/Jurassic boundary, 208 million years ago, is associated with widespread extinctions in both the marine and terrestrial biota. The cause of these extinctions has been widely attributed to the eruption of flood basalts of the Central Atlantic Magmatic Province. This volcanic event is thought to have released significant amounts of CO2 into the atmosphere, which could have led to catastrophic greenhouse warming, but the evidence for CO2-induced extinction remains equivocal. Here we present the carbon isotope compositions of pedogenic calcite from palaeosol formations, spanning a 20-Myr period across the Triassic/Jurassic boundary. Using a standard diffusion model, we interpret these isotopic data to represent a rise in atmospheric CO2 concentrations of about 250 p.p.m. across the boundary, as compared with previous estimates of a 2,000-4,000 p.p.m. increase. The relative stability of atmospheric CO2 across this boundary suggests that environmental degradation and extinctions during the Early Jurassic were not caused by volcanic outgassing of CO2. Other volcanic effects-such as the release of atmospheric aerosols or tectonically driven sea-level change-may have been responsible for this event. PMID- 11395766 TI - A phenomenological model for precursor volcanic eruptions. AB - Intense explosions of relatively short duration frequently precede large explosive and effusive volcanic eruptions-by as much as weeks to months in the case of very viscous magmas. In some cases, such pre-eruption activity has served as a sufficient warning to those living in the vicinity to evacuate and avoid calamity. Precursor events seem to be related to the formation of a magma pathway to the surface, but their precise interpretation is a long-standing puzzle. It has been inferred from theoretical studies that exsolution of volatiles might create an almost separate gas pocket at the tip of a propagating dyke. Here we explain the role that such a process may have, using a laboratory study of the transient propagation of a liquid-filled crack with a gas pocket at its tip that grows with time. We show that once the gas pocket acquires sufficient buoyancy to overcome the fracture resistance of the host solid the dynamics of the gas pocket, rather than those of the liquid, determine the velocity of the crack tip. Furthermore, we find that the gas can ultimately separate from the liquid. We propose that fast-moving, gas-rich pockets reaching the surface ahead of the main liquid-filled fissure could be the origin of many precursor eruptions. PMID- 11395767 TI - Reykjanes "V"-shaped ridges originating from a pulsing and dehydrating mantle plume. AB - Prominent crustal lineations straddle the Reykjanes ridge, south of Iceland (Fig. 1). These giant V-shaped features are thought to record temporal variations in magma production at the Reykjanes ridge axis, associated with along-axis flow of Icelandic plume material. It has been proposed that this flow is channelled preferentially along the ridge axis, and that temporal variability is induced by fluctuations of the Iceland plume itself or, alternatively, by relocations of the ridge axis on Iceland. Here I present a geodynamic model that predicts the formation of crustal V-shaped ridges from a pulsing and radially flowing mantle plume. In this model, plume pulses produce mantle temperature perturbations that expand away from the plume in all directions beneath the zone of partial melting. The melting zone has a high viscosity owing to mantle dehydration at the onset of partial melting. This high-viscosity region allows for reasonable variations in crustal thickness, produces crustal Vs that extend hundreds of kilometres along the axis, and prevents the plume material from being preferentially channelled along the ridge axis. The angle of the crustal V-shaped features relative to the ridge axis reflects the rate of lateral plume flow, which remains several times greater than the ridge half-spreading rate over the length of a crustal V. Consequently, this radially expanding plume produces lineations in crustal thickness and free-air gravity anomalies that appear to be nearly straight. PMID- 11395768 TI - Earliest evidence for efficient oral processing in a terrestrial herbivore. AB - Herbivores can increase their digestion rate by mechanically reducing particle size through oral trituration. Groups of terrestrial vertebrates with the greatest capacity to reduce tough plant foods orally are also the most abundant and diverse, as exemplified by ornithopod dinosaurs during the Mesozoic and extant artiodactyl and perissodactyl mammals. Thus, the effective oral processing of high-fibre plant material seems to represent an evolutionary innovation of both functional and macroevolutionary significance. However, evidence for oral processing is poorly documented in the fossil record, especially during the initial stages of terrestrial vertebrate diversification. Here we report on the basal anomodont Suminia getmanovi, the only known Palaeozoic vertebrate in which unequivocal specializations in its cranium and teeth for high-fibre herbivory are well preserved. We propose that the capacity to comminute tough plant foods was critical to the diversification of anomodonts, the most diverse, widely dispersed and abundant group of Palaeozoic terrestrial vertebrates, and to the onset of modern terrestrial ecosystems. PMID- 11395769 TI - Effects of macrophyte species richness on wetland ecosystem functioning and services. AB - Wetlands provide many important ecosystem services to human society, which may depend on how plant diversity influences biomass production and nutrient retention. Vascular aquatic plant diversity may not necessarily enhance wetland ecosystem functioning, however, because competition among these plant species can be strong, often resulting in the local dominance of a single species. Here we have manipulated the species richness of rooted, submerged aquatic plant (macrophyte) communities in experimental wetland mesocosms. We found higher algal and total plant (algal plus macrophyte) biomass, as well as lower loss of total phosphorus, in mesocosms with a greater richness of macrophyte species. Greater plant biomass resulted from a sampling effect; that is, the increased chance in species mixtures that algal production would be facilitated by the presence of a less competitive species-in this case, crisped pondweed. Lower losses of total phosphorus resulted from the greater chance in species mixtures of a high algal biomass and the presence of sago pondweed, which physically filter particulate phosphorus from the water. These indirect and direct effects of macrophyte species richness on algal production, total plant biomass and phosphorus loss suggest that management practices that maintain macrophyte diversity may enhance the functioning and associated services of wetland ecosystems. PMID- 11395770 TI - Sexual selection and the maintenance of sex. AB - Sex is expensive. A population of females that reproduce asexually should prima facie have twice the growth rate of an otherwise equivalent anisogamous sexual population lacking paternal care, or a population with modes of paternal care that can be co-opted by parthenogenetic females. The two leading theories for the maintenance of sex require either synergistic interactions between deleterious mutations, or antagonistic epistasis between beneficial mutations. Current evidence is equivocal as to whether the required levels of epistasis exist. Here I show that a third factor, differential male mating success (or, more generally, higher variance in male than in female fitness), can drastically reduce mutational load in sexual populations with or without any form of epistasis. Differential mating success has the further advantage of being ubiquitous, and is likely to have preceded or evolved concurrently with anisogamy. PMID- 11395771 TI - Sexual selection and the maintenance of sexual reproduction. AB - The maintenance of sexual reproduction is a problem in evolutionary theory because, all else being equal, asexual populations have a twofold fitness advantage over their sexual counterparts and should rapidly outnumber a sexual population because every individual has the potential to reproduce. The twofold cost of sex exists because of anisogamy or gamete dimorphism-egg-producing females make a larger contribution to the zygote compared with the small contribution made by the sperm of males, but both males and females contribute 50% of the genes. Anisogamy also generates the conditions for sexual selection, a powerful evolutionary force that does not exist in asexual populations. The continued prevalence of sexual reproduction indicates that the 'all else being equal' assumption is incorrect. Here I show that sexual selection can mitigate or even eliminate the cost of sex. If sexual selection causes deleterious mutations to be more deleterious in males than females, then deleterious mutations are maintained at lower equilibrium frequency in sexual populations relative to asexual populations. The fitness of sexual females is higher than asexuals because there is no difference in the fecundity of sexual females and asexuals of the same genotype, but the equilibrium frequency of deleterious mutations is lower in sexual populations. The results are not altered by synergistic epistasis in males. PMID- 11395772 TI - PCR amplification of the Irish potato famine pathogen from historic specimens. AB - Late blight, caused by the oomycete plant pathogen Phytophthora infestans, is a devastating disease of potato and was responsible for epidemics that led to the Irish potato famine in 1845 (refs 1,2,3,4,5). Before the 1980s, worldwide populations of P. infestans were dominated by a single clonal lineage, the US-1 genotype or Ib mitochondrial DNA (mtDNA) haplotype, and sexual reproduction was not documented outside Mexico, the centre of diversity of the pathogen. Here we describe the amplification and sequencing of 100-base-pair fragments of DNA from the internal transcribed spacer region 2 from 28 historic herbarium samples including Irish and British samples collected between 1845 and 1847, confirming the identity of the pathogen. We amplified a variable region of mtDNA that is present in modern Ib haplotypes of P. infestans, but absent in the other known modern haplotypes (Ia, IIa and IIb). Lesions in samples tested were not caused by the Ib haplotype of P. infestans, and so theories that assume that the Ib haplotype is the ancestral strain need to be re-evaluated. Our data emphasize the importance of using historic specimens when making inferences about historic populations. PMID- 11395773 TI - Retinal ganglion cells act largely as independent encoders. AB - Correlated firing among neurons is widespread in the visual system. Neighbouring neurons, in areas from retina to cortex, tend to fire together more often than would be expected by chance. The importance of this correlated firing for encoding visual information is unclear and controversial. Here we examine its importance in the retina. We present the retina with natural stimuli and record the responses of its output cells, the ganglion cells. We then use information theoretic techniques to measure the amount of information about the stimuli that can be obtained from the cells under two conditions: when their correlated firing is taken into account, and when their correlated firing is ignored. We find that more than 90% of the information about the stimuli can be obtained from the cells when their correlated firing is ignored. This indicates that ganglion cells act largely independently to encode information, which greatly simplifies the problem of decoding their activity. PMID- 11395774 TI - Regulation of Ca2+ channel expression at the cell surface by the small G-protein kir/Gem. AB - Voltage-dependent calcium (Ca2+) channels are involved in many specialized cellular functions, and are controlled by intracellular signals such as heterotrimeric G-proteins, protein kinases and calmodulin (CaM). However, the direct role of small G-proteins in the regulation of Ca2+ channels is unclear. We report here that the GTP-bound form of kir/Gem, identified originally as a Ras related small G-protein that binds CaM, inhibits high-voltage-activated Ca2+ channel activities by interacting directly with the beta-subunit. The reduced channel activities are due to a decrease in alpha1-subunit expression at the plasma membrane. The binding of Ca2+/CaM to kir/Gem is required for this inhibitory effect by promoting the cytoplasmic localization of kir/Gem. Inhibition of L-type Ca2+ channels by kir/Gem prevents Ca2+-triggered exocytosis in hormone-secreting cells. We propose that the small G-protein kir/Gem, interacting with beta-subunits, regulates Ca2+ channel expression at the cell surface. PMID- 11395775 TI - KANADI regulates organ polarity in Arabidopsis. AB - Leaves and floral organs are polarized along their adaxial-abaxial (dorsal ventral) axis. In Arabidopsis, this difference is particularly obvious in the first two rosette leaves, which possess trichomes (leaf hairs) on their adaxial surface but not their abaxial surface. Mutant alleles of KANADI (KAN) were identified in a screen for mutants that produce abaxial trichomes on these first two leaves. kan mutations were originally identified as enhancers of the mutant floral phenotype of crabs claw (crc), a gene that specifies abaxial identity in carpels. Here we show that KAN is required for abaxial identity in both leaves and carpels, and encodes a nuclear-localized protein in the GARP family of putative transcription factors. The expression pattern of KAN messenger RNA and the effect of ectopically expressing KAN under the regulation of the cauliflower mosaic virus (CAMV) 35S promoter indicate that KAN may also specify peripheral identity in the developing embryo. PMID- 11395776 TI - Role of PHABULOSA and PHAVOLUTA in determining radial patterning in shoots. AB - The upper side of the angiosperm leaf is specialized for efficient capture of sunlight whereas the lower side is specialized for gas exchange. In Arabidopsis, the establishment of polarity in the leaf probably requires the generation and perception of positional information along the radial (adaxial versus abaxial or central versus peripheral) dimension of the plant. This is because the future upper (adaxial) side of the leaf develops from cells closer to the centre of the shoot, whereas the future under (abaxial) side develops from cells located more peripherally. Here we implicate the Arabidopsis PHABULOSA and PHAVOLUTA genes in the perception of radial positional information in the leaf primordium. Dominant phabulosa (phb) and phavoluta (phv) mutations cause a dramatic transformation of abaxial leaf fates into adaxial leaf fates. They do so by altering the predicted sterol/lipid-binding domains of ATHB14 and ATHB9, proteins of previously unknown function that also contain DNA-binding motifs. This change probably renders the protein constitutively active, implicating this domain as a central regulator of protein function and the PHB and PHV proteins as receptors for an adaxializing signal. PMID- 11395777 TI - Defects in mismatch repair promote telomerase-independent proliferation. AB - Mismatch repair has a central role in maintaining genomic stability by repairing DNA replication errors and inhibiting recombination between non-identical (homeologous) sequences. Defects in mismatch repair have been linked to certain human cancers, including hereditary non-polyposis colorectal cancer (HNPCC) and sporadic tumours. A crucial requirement for tumour cell proliferation is the maintenance of telomere length, and most tumours achieve this by reactivating telomerase. In both yeast and human cells, however, telomerase-independent telomere maintenance can occur as a result of recombination-dependent exchanges between often imperfectly matched telomeric sequences. Here we show that loss of mismatch-repair function promotes cellular proliferation in the absence of telomerase. Defects in mismatch repair, including mutations that correspond to the same amino-acid changes recovered from HNPCC tumours, enhance telomerase independent survival in both Saccharomyces cerevisiae and a related budding yeast with a degree of telomere sequence homology that is similar to human telomeres. These results indicate that enhanced telomeric recombination in human cells with mismatch-repair defects may contribute to cell immortalization and hence tumorigenesis. PMID- 11395778 TI - A freely diffusible form of Sonic hedgehog mediates long-range signalling. AB - The secreted protein Sonic hedgehog (Shh) exerts many of its patterning effects through a combination of short- and long-range signalling. Three distinct mechanisms, which are not necessarily mutually exclusive, have been proposed to account for the long-range effects of Shh: simple diffusion of Shh, a relay mechanism in which Shh activates secondary signals, and direct delivery of Shh through cytoplasmic extensions, termed cytonemes. Although there is much data (using soluble recombinant Shh (ShhN)) to support the simple diffusion model of long-range Shh signalling, there has been little evidence to date for a native form of Shh that is freely diffusible and not membrane-associated. Here we provide evidence for a freely diffusible form of Shh (s-ShhNp) that is cholesterol modified, multimeric and biologically potent. We further demonstrate that the availability of s-ShhNp is regulated by two functional antagonists of the Shh pathway, Patched (Ptc) and Hedgehog-interacting protein (Hip). Finally, we show a gradient of s-ShhNp across the anterior-posterior axis of the chick limb, demonstrating the physiological relevance of s-ShhNp. PMID- 11395783 TI - New directions in mental health public policies in Brazil. PMID- 11395784 TI - [Determinant factors of postpartum weight retention: a literature review]. AB - This study is a literature review of factors determining postpartum weight gain retention. According to the literature, the main factors include gestational weight gain and intensity of lactation. Various studies show that the greater the weight gain during pregnancy, the greater the postpartum weight retention. Smoking was associated with lower retention, while lactation only demonstrated a protective effect against weight retention in some studies. Various factors, like physical activity, food consumption, age at menarche, number of abortions, and mother's work status have still not been properly studied. The conclusion is that there are few studies on postpartum weight changes, suggesting the need for new research on the theme, especially in developing countries where obesity has been insufficiently researched thus far. PMID- 11395785 TI - [Molecular biomarkers in cancer: implications for epidemiological research and public health]. AB - Identification of molecular biomarkers is a common result of current cancer epidemiological research. Both genetic and molecular epidemiology have enjoyed impressive developments in recent decades, with important repercussions on traditional epidemiological approaches. In this paper we evaluate the new frontiers of cancer epidemiology, incorporating both genetic and molecular biology approaches. We examine the current knowledge of molecular biomarkers for exposure and susceptibility to cancer, the role of gene mutations in carcinogenesis, and their application to epidemiological studies. By exploring the status of relevant biomarkers, these approaches become effective in evaluating exposure and susceptibility and show enormous potential for elucidating mechanisms of carcinogenesis and the effect of risk factors in cancer. However, these approaches are necessarily more invasive and raise several ethical issues for consideration by both researchers in public health and society as a whole. PMID- 11395786 TI - ["Knowing about AIDS" and sexual precautions among low-income women from the southern area of Buenos Aires. Notes for defining prevention policies]. AB - This study is part of a line of research on gender and prevention in a research program on the social construction of HIV/AIDS. We present the results of an ethnographic study among low-income women 15-35 years old in the southern area of Buenos Aires. The area has the highest number of HIV/AIDS cases and high poverty levels, extensive social degradation, and urban violence. According to our results, in the interface between "knowing about" and "behaving" there are complex processes involving stigmatized and gender-biased representations of HIV/AIDS as "other people's problem" and social and sexual relations permeated by gender stereotypes and roles. We believe that planning of prevention should be based on the consideration of overall social practices and specifically the characteristics of gender relations, prioritizing relational strategies between women and men and promoting critical reflection on the main nodes organizing daily life and active participation in the production of social relations and practices of reciprocity and equity. The increasingly precarious conditions in social life intensifies poor women's vulnerability and social interaction contexts that relate to the socioeconomic and symbolic role played by women. PMID- 11395787 TI - Is pertussis actually reemerging? Insights from an individual-based model. AB - In this paper, we introduce a spatially explicit, individual-based model developed to simulate the dynamics of pertussis in a small population. With this simulation approach, complex epidemic systems can be built using information on parasite population structure (strain diversity, virulence diversity, etc.), human population structure (individual risk, age structure, interaction matrices, immune response, etc.), as well as mechanisms of evolution and learning. We parameterized our model to describe pertussis in an age-structured community. Pertussis or whooping cough is an acute infection of the respiratory tract caused by Bordetella pertussis. Despite wide-scale vaccination in many countries, this disease is reemerging throughout the world in both adults and children. Emergence has been explained by many factors: wane of vaccine and natural immunity, increase of asymptomatic carriers, and/or natural selection of non-vaccine strains. Here, we model these hypotheses and analyze their potential impact on the observed increase of pertussis notification. PMID- 11395788 TI - [Use of dust and air as indicators of environmental pollution in areas adjacent to a source of stationary lead emission]. AB - This study measured lead concentrations in both the outdoor air and household dust from houses located around a lead-acid battery repair shop. Such installations are one of the largest sources of lead exposure, since outdated technology is still used, coupled with the lack of strict air-quality control programs. Measurements of the air lead concentration around the repair shop were carried out at 6 points, approximately 25 and 500 m from the shop. Over 50% of the air samples exceeded the limit of 1.5 microg Pb.m-3 (range 0.03 - 183.3 microg Pb.m-3). House dust samples were collected from 6 places in houses located at approximately 25, 50, and 500 m from the repair shop, and the concentration of 1,500 microg Pb.m-2 for lead in house dust was exceeded in 44% of the samples, with results varying from 2.2 to 54,338.9 microg Pb.m-2. PMID- 11395789 TI - [Cesarean sections: an epidemic profile]. AB - This study characterizes cesarean rates among women giving birth in Sao Jose do Rio Preto in 1992. Household interviews were conducted with 553 women selected through records on live births. Estimated incidence of cesarean sections was 80.5%, highlighting the procedure's widespread use in the study sample. Confirming a prior hypothesis, cesarean rates increased with parity and were statistically associated with age and tubal ligation. As for socioeconomic status, cesarean rates increased according to income and were higher among women using private health care (with or without health insurance). Interviews with professors in the local medical school and a related survey of the local media showed an increase in the valorization of cesareans. Confirming this trend, women interviewed in the survey expressed their own active valorization of this procedure. PMID- 11395790 TI - [Reliability of information on the underlying cause of death from external causes in people under 18 years of age in the Municipality of Duque de Caxias, Rio de Janeiro, Brazil]. AB - This study assesses the quality of official data on mortality due to external causes. It examines the agreement in the underlying cause of death by accidents and violence in children and adolescents under 18 years of age in the Municipality of Duque de Caxias, Rio de Janeiro State. Codings based on death certificates (DC) issued by the Institute of Forensic Medicine - IFM (ICDifm) and those complemented by the Municipal Secretariat of Health - MSH (ICDmsh) are compared to the coding directly obtained from autopsy logs and attached to police reports accessible at the IFM (ICDresearch). Levels of agreement (kappa) between ICDmsh/ICDifm, ICDresearch/ ICDifm, and ICDresearch/ICDmsh were 0.33, 0.26, and 0.81, respectively, in 1995 and 0.26, 0.22, and 0.71 in 1996. When analyzed together, all three sources agreed by 45% and 37.6% in the respective years. ICDresearch and ICDmsh agreed among themselves and disagreed with ICDifm by 46.9% in 1995 and 48.2% in 1996. Results show that the IFM fails to transfer available information to the MSH via DCs. Although a satisfactory updating is subsequently carried out at the MSH, the relevance and legality of this procedure should be called into question and further discussed. PMID- 11395791 TI - [Equity in socioeconomic areas with impact on health in countries of European Union]. AB - It may sometimes be more appropriate to correct inequalities in the domains of education, lifestyles, or nutrition than that of health. The current study proposes to measure existing inequalities in European Union member countries in socioeconomic areas with an impact on the population's health. The Gini coefficient was used to quantify inequalities among member countries with regard to education and cultural activities, lifestyles, nutrition, unemployment, use of health care services, and health expenditures. The indicators with the highest Gini coefficients are the percentage of the population with a secondary school education (level of schooling just preceding the university), per capita butter consumption, and per capita consumption of packaging for medications. It is cause for concern to the health sector that education displays such great inequality in the percentage of individuals enrolled in secondary education within the European Union. Existing inequality in per capita butter consumption is equally important due to the impact nutrition has on the incidence of cardiovascular and cerebral vascular diseases, for example, and the weight these diseases have on the mortality profile. PMID- 11395792 TI - [Biological monitoring: concepts and applications in public health]. AB - This study provides an overview of the theoretical discussion on potential uses for biological monitoring of exposure to chemical substances as related to human health, considering different concepts: definitions, uses, and limitations of internal dose and biological effect indicators and their availability for the substances to be quantified; knowledge of reference values, action levels, and limits based on health and negotiated patterns in biological monitoring interpretation and perspectives; and ethical and social problems in practice and within different preventive practices and their use in public health. Biological monitoring is the result of an exposure situation with conclusions based on scientific and consensus values, rules, and legislation. Biological monitoring as a continuous process and related to actually observed cases has helped establish technological exposure reference values and consensus levels as indicators for improving the environment and the workplace. As a step in the decision-making process in risk analysis, biological monitoring needs to be critically assessed as to its ethical aspects in light of the end use of results and values, which are references for application of this methodology. PMID- 11395793 TI - [Infant mortality and living conditions: the reproduction of social inequalities in health during the 1990s]. AB - An ecological study was conducted to determine the infant mortality trend from 1991 to 1997 and to analyze its relationship to living conditions in Salvador, Bahia State, Brazil. Inequality patterns in infant death were analyzed by spatial distribution and a compound socioeconomic index. The data showed a decline in the infant mortality rate, with neonatal deaths and perinatal causes playing a growing role. Despite this overall trend, the infant mortality rate increased in 1992, and it was only in 1997 that it returned to the 1991 level. This fact was interpreted as related to worsening living conditions during that period. Spatial distribution highlights the persistence of health inequalities; education was the variable with the most significant correlation rate. When distributed according to the living conditions index (LCI), both the infant mortality rate and proportional infant mortality showed a linear increase from the intermediate stratum (20.4 per mil ) to the lowest (29.3 per mil ) and from the highest stratum (5.3%) to the lowest (13.3%), respectively. The authors conclude that despite the reduction in the total infant mortality rate, the persistence of social inequalities and a social process that hinders improvement of living conditions are responsible for the inequalities observed in infant mortality. PMID- 11395794 TI - [Integration, exclusion and solidarity in the current social policy debate]. AB - The current global trend involving transformation of work and the crisis in large government economic development and intervention projects in the market are raising new challenges for social policies to combat poverty. This study reconstitutes and analyzes the tensions, paradoxes, and main consequences of this process, focusing on problems concerned with combining solidarity strategies and public welfare systems. PMID- 11395795 TI - [Energy adequacy ratio (intake/requirements) as an indicator of families nutritional assessment: a critical analysis of methods applied to food consumption surveys]. AB - The authors discuss problems associated with the use of the energy adequacy ratio as an indicator of nutritional status in population-based studies derived from household food consumption surveys. They describe the use of food consumption data to estimate family energy intake and to compare it among families and with family requirements to determine the family energy adequacy ratio. The article also discusses the use of international recommendations for human energy requirements and their limitations, based on the lack of consideration of non food conditions, adaptive mechanisms, inadequacy of estimates of the baseline metabolism rate, and the lack of information to evaluate energy expenditure in non-work physical activities in the household surveys consulted. To illustrate the discussion, the authors use data from the National Family Budget Survey (ENDEF) conducted by the Brazilian Census and Statistics Bureau (IBGE) in 1974 1975. The article concludes that the main use of the adequacy ratio is to identify families suffering food restriction. PMID- 11395796 TI - [Community genetics and hemophilia in a Brazilian population]. AB - Hemophilia is an important hemorrhagic disease in Brazil, affecting about 1 out of every 10,000 males. Patient's self-perception of hemophilia and interaction with the community are relevant to the clinical management of this disease. We investigated several social, psychological, and community aspects of hemophilia in a Brazilian population (Campinas, Sao Paulo State), interviewing 30 hemophiliac males, a control sample comprised of 73 non-hemophiliac brothers, and 641 individuals from the community. According to our results, more severe social disability in the hemophiliac patient was related to economic factors, mainly unemployment; however, no difference was found in relation to marital status, reproduction, or education. Self-perception of changes in health and lifestyle by individuals with hemophilia showed frequent self-stigmatization, along with depression, anxiety, and insecurity. The community showed a widespread lack of familiarity with hemophilia (49%), viewing people with hemophilia with the kinds of prejudices often observed in relation to people with infectious diseases, like AIDS. The paper concludes by recommending that a community-based program be implemented to improve the social adjustment status of individuals with hemophilia. PMID- 11395797 TI - [Mental health and work: a discussion on the connection between work and diagnosis, based on daily practice]. AB - Seven cases of workers selected among 150 who had undergone psychiatric treatment at the Workers' Health Reference Centers in Santo Amaro and Andre Gabois from 1994 to 1997 were presented for a detailed analysis characterizing work situations and discussing definition of the diagnosis and causal connections with work. The qualitative case study provides a thematic analysis of the patient history and files. Work situations are characterized by: unhealthy working conditions, problems related to work organization, inadequate human resources management, and urban violence. Symptoms included: fear, anxiety, depression, nervousness, tension, fatigue, malaise, loss of appetite, sleep disorders, and psychosomatic disorders (gastritis, hypertension); in addition, workers were unable to forget about work while off the job. Diagnosis varied. Three cases involved post-traumatic syndrome. Two cases involved organic psychosis linked to accidents or exposure to neurotoxic chemical products. Cases also included neurotic syndromes of fatigue, depression, and paranoia, as well as adaptation and reaction to acute stress. In all of the cases it was possible to relate the clinical picture to the work situation. PMID- 11395798 TI - [Death certificates as a marker for under-recording of AIDS cases]. AB - This study aimed to estimate and evaluate the under-recording of AIDS cases that evolved to death. From 1991 to 1995, the Mortality Information System recorded 9,213 adult deaths due to AIDS, related by name, date of birth, and date of death to 15,505 AIDS recorded by the Disease Data Registration System from 1982 to 1996. This procedure showed that 51.9% of AIDS deaths recorded in the period were not recorded as AIDS cases as they should have been. Univariate and multivariate analysis showed that women had a greater chance of under-recording than men (OR = 1.27). A lower probability of proper recording was observed in individuals with less schooling, and illiterates had a two-fold chance of not being properly recorded, as compared to individuals with college degrees. Deaths that occurred in health facilities classified as private units had a greater probability of not being recorded than those occurring elsewhere (mainly those classified as public reference units) (OR = 2.58). Deaths occurring in the city of Rio de Janeiro had a greater probability of under-recording than those in other cities (OR = 2.20). PMID- 11395799 TI - [Cost variation of hospitalization due to injuries: head trauma and fire arm accidents]. AB - This article proposes two linear regression models on cost variation of hospitalization (based on data from the Brazilian National Unified Health System SUS) for two types of injuries: head trauma and related cases (n = 98,156); fire arm injuries and related cases (n = 8,970). Data were collected from the 1997 standardized Hospital Admittance Forms covering all of Brazil. Explanatory variables were gender, age, hospital administration model (public, private, etc.), region of the country, hospitalization in a capital city, use of ICU, surgery, death, duration of hospitalization, most frequent procedures, special procedures, and interaction among certain variables. The two models adjusted well, with R2 = 0.7264 for the first and 0.7663 for the second. Explanatory variables in the first model were all significant, and only three variables in the second failed to show statistical significance. The two main variables in both models were use of ICU and surgery. Diagnostics for detection of outliers, multicolinearity, model specification error, and homoscedasticity were performed. PMID- 11395800 TI - [Gender and night work: sleep, daily life and the experiences of night shift workers]. AB - This paper deals with the impact of night work from a gender perspective, through a field study at a factory employing men and women on the night shift. It is based on data for hours of sleep over the course of several weeks, socio demographic data, and job information, using a semi-structured interview. The methodology includes chronobiological aspects of sleep (mainly quantitative data) and workers' discourse concerning gender-related experiences in the day-to-night switch. Despite gender issues and differences in daily life, the switch is perceived quite vividly by both men and women, permeating several aspects of life, like health, leisure, studies, and intimate personal relations. Quantitative sleep analysis showed more severe effects of night work on women, especially those with children. Such sleep patterns were associated with different expectations among men and women, revealing gender issues that are essential for understanding the reality of individuals who work odd hours. PMID- 11395801 TI - [The importance of water testing for public health in two regions in Rio de Janeiro: a focus on fecal coliforms, nitrates, and aluminum]. AB - In developing countries, due to poor sanitation conditions and poor quality of drinking water, typical water-borne diseases and more recently diseases caused by drinking water with high concentrations of nitrates and certain metals like aluminum have increased the concern over the health effects of these compounds. Several articles have shown associations between nitrates and methemoglobinemia in children, and aluminum and Alzheimer disease in adults. This study identified water quality with several parameters in non-conformity with Brazilian drinking water standards (Ruling 36/90): more than 50% of all samples from both regions contained fecal coliforms; some 31% of water samples from wells in Duque de Caxias had excessive nitrate concentrations; 100% of all groundwater samples from both regions showed aluminum concentrations not conforming to the norm, with the same result for 100% of samples from the drinking water distribution system in Sao Goncalo and 75% of same in Duque de Caxias. This lack of conformity poses several health risks for the local population. PMID- 11395802 TI - [The Bambui Project: a population-based study of factors associated with regular utilization of dental services in adults]. AB - A cross-sectional study was conducted in Bambui, Minas Gerais, to identify factors associated with regular use of dental services. Participants were interviewed with a structured questionnaire and previously validated questions. 999/1,221 (81.8%) randomly selected individuals aged > 18 years participated in the Bambui dental survey. Of these, 654 out of 656 individuals who had at least one natural tooth and had visited a dentist during their lifetime participated in the study. Results adjusted by multiple logistic regression showed that regular use of dental services was significantly related to having > 8 and 4-7 years of schooling (OR = 9.90; 95% CI = 2.90-33.77 and OR = 3.87; 95% CI = 1.11-13.51, respectively), having a preference for restorative treatment rather than extraction (OR = 4.91; 95% CI = 2.23-10.79), having no present need of dental treatment (OR = 4.87; 95% CI = 3.17-7.49), and belief that visiting the dentist prevents tooth decay and gum disease (OR = 1.73; 95% CI = 1.13-2.65). The results show that regular use of dental services was related to factors distributed in the Andersen and Newman model (1973) explaining use of dentistry services. PMID- 11395803 TI - [Male homicide in the Metropolitan Region of Sao Paulo between 1979 and 1998: a pictorial approach]. AB - Homicide has become a major social problem in Brazil in the last two decades. In Greater Metropolitan Sao Paulo, Southeast Brazil, the proportional incidence of male homicides increased from 35 to 121 (x 10-5) from 1979 to 1998. This study presents the trend in this phenomenon using a set of three-dimensional figures. We show both the absolute number and proportional incidence of male deaths by homicide and the natural logarithm of the male/female relative risk of homicide for Greater Metropolitan Sao Paulo from 1979 to 1998. Seven 3D figures show the homicide trend by age, sex, and year. The figures show the main trends in homicide distribution in the male population during the study period: a clear predominance among adolescents and young adults, a systematic and sharp increase in that same age range, a systematic spread to the lower (below-18) and higher (above-30) age groups, and a non-negligible incidence above the age of 40. PMID- 11395804 TI - [The Helsinki Declaration: relativism and vulnerability]. AB - The Helsinki Declaration is a crucial ethical landmark for clinical research involving human beings. Since the Declaration was issued, a series of revisions and modifications have been introduced into the original text, but they have not altered its humanist approach or its international force for regulating clinical research. A proposal for an extensive revision of the Declaration's underlying ethical principles has been debated for the past four years. If the proposal is approved, international clinical research involving human beings will be modified, further increasing the vulnerability of certain social groups. This article discusses the historical process involved in passing the Helsinki Declaration and the most recent debate on the new draft. The article analyzes the new text's social implications for underdeveloped countries, arguing for a political approach to the vulnerability concept. PMID- 11395805 TI - [Public and occupational health aspects related to municipal solid waste management]. AB - Based on a literature review, the paper calls attention to public and occupational health problems related to poor municipal solid waste management resulting from a development model that views environmental protection, public health, and workers' health as secondary issues. The authors emphasize the need for research to support a new waste management model highlighting the importance of environmental preservation and protection of human health, in addition to suggesting measures to achieve such goals. The discussion aims to incorporate solid waste management into the public health agenda. PMID- 11395806 TI - [Epidemiological research trends in chronic diseases]. AB - Identification of adult risk factors has contributed to a decrease in the occurrence of cardiovascular diseases and some types of cancer, particularly in developed countries. However, the prevailing etiologic model emphasizes risk factors associated with life style and occupational exposures. This emphasis leads to an excessive individualization of risk, weakening the link between epidemiology and public health. This article comments on the opinions of epidemiologists concerning the failures, limitations, and evolution of modern epidemiology that have helped redirect the field's theoretical underpinnings. Alternatives to the current model tend to integrate findings from molecular biology with classic risk factors and early adverse conditions in life, taking into account all these causal influences in hierarchical levels of organization. The author identifies and comments on three important approaches in this evolution: molecular epidemiology, the theory of early life factors, and multilevel models. PMID- 11395807 TI - Considerations on methodology used in the World Health Organization 2000 Report. AB - The article analyzes the World Health Organization Report for 2000, with emphasis placed on the methodology used to analyze the indicators utilized to compare and classify the performance of the health systems of the 191 member countries. The Report's contribution was the compromise of monitoring the performance of the health systems of member countries, but because of the inconsistent way it was elaborated, and the utilization of questionable scientific evaluation methodologies, the Report fails to give a clear picture. A criterion-based methodology revision is imposed. The main problems in evidence are the choice of individual indicators of disparity in health that discount the population profile, the inadequate control of the impact of social disparities over the performance of the systems, the evaluation of the responsibility of systems that are only partially articulated to the right of the citizens, the lack of data for a great number of countries, consequently having inconsistent estimations, and the lack of transparency in the methodological procedures in the calculation of some indicators. The article suggests a wide methodological revision of the Report. PMID- 11395808 TI - [Antimicrobial resistance of coliform isolates from expressed human milk]. AB - The dispersion of potentially pathogenic, antibiotic-resistant microorganisms via expressed human milk can be considered a risk factor. The aim of this study was to contribute to a better understanding of coliform isolates from expressed human milk and their antimicrobial resistance profiles. The sampling scheme followed a totally randomized design, using 837 samples of expressed human milk. Of these, 71 (8.48%) were identified as contaminated with total coliforms, although in none of the samples did the population exceed 1.0x10(3) MPN/ml. Most of the microorganisms isolated (91.6%) belonged to only two species, Enterobacter cloacae and Klebsiella pneumoniae, which when subjected to antibiograms, revealed that several strains showed prior resistance to some of the antimicrobials tested. Coliforms may grow in expressed human milk if it is improperly stored, depleting protection factors and reducing the milk's nutritional value. PMID- 11395809 TI - [Training for diagnosis of intestinal parasitic diseases in the national laboratory system of Cuba]. AB - A national training project in the diagnosis of intestinal parasites was conducted in 1997. An initial national course was followed respectively by courses in the Central, Eastern, and Western Provinces. Our results showed that Cryptosporidium parvum, Cyclospora cayetanensis, and leukocytes showed a significantly lower percentage of errors after the training than before (p < 0.01). The same occurred with Entamoeba histolytica/E. dispar and Chilomastix mesnilii (p < 0.05). Among the helminths, Taenia spp., Fasciola hepatica, and hookworm showed significantly fewer errors after the training (p < 0.01). In the other specimens, few mistakes were found both at the beginning and after the training, and the percentage of errors did not change (p > 0.05). Furthermore, when comparing scores before and after training, a significant increase in median scores appeared in the Central Provinces (p < 0.05), Western Provinces (p < 0.01), and in the entire series (p < 0.01). The results showed the effectiveness of this intervention; these periodic mandatory training courses should be developed together with national programs for quality assessment in Parasitology. PMID- 11395810 TI - [An outbreak of acute schistosomiasis at Porto de Galinhas beach, Pernambuco, Brazil]. AB - We recently confirmed several cases of acute schistosomiasis in Porto de Galinhas beach, Northeast Brazil. A total of 662 patients were diagnosed by parasitological and clinical examinations. The infection likely occurred during the September 7 national holiday, when heavy rainfall flooded the Ipojuca River and people were infected when the water covered their yards. Families were continuously exposed to infection for a period of three weeks until the water had completely dried up. Previous investigation suggests that snail vectors were introduced as a result of landfill in marshy areas. The swamp-flooding of such areas facilitated the emergence of slums surrounded by snail breeding sites. Heavy rainfall caused open-air sewage ditches to overflow, allowing for infection of snails by Schistosoma mansoni. Thus, continuous floods were responsible for the spread of human infection. Clinical and laboratory results identified 62% of acute cases of S. mansoni. Complementary studies are being conducted to define the impact and epidemiological meaning of the acute schistosomiasis outbreak. PMID- 11395811 TI - Scientific electronic publication: An updated report. PMID- 11395812 TI - One world, one people, one surgery. PMID- 11395814 TI - Quality of life before and after laparoscopic Nissen fundoplication. AB - BACKGROUND: Gastroesophageal reflux disease (GERD) is a common disorder in the Western world. The acute disease can usually be managed by medical therapy. To prevent relapse, many patients require lifelong medication. In these patients, laparoscopic antireflux surgery offers a good alternative. The aim of this study was to evaluate the postoperative results and compare pre- and postoperative quality of life after laparoscopic Nissen fundoplication. METHODS: Clinical investigations, including esophageal manometry, pH monitoring, and endoscopy, and a previously validated Quality of Life Index, were performed before and a median of 41 month after antireflux surgery in 75 patients. RESULTS: After laparoscopic Nissen fundoplication, the percentage of total time with pH <4 decreased from 10.4% to 3.2% on 24-h pH monitoring. The mean pressure of the lower esophageal sphincter improved from 8.1 to 12.3 mmHg. Esophagitis healed in 63 of 66 patients in whom it was present prior to surgery. The overall Quality of Life Index improved significantly from 86 +/- 16 to 116 +/- 16. CONCLUSION: Laparoscopic fundoplication provides effective and durable relief of reflux in patients with GERD. The Quality of Life Index showed significant improvement after surgery. PMID- 11395813 TI - Complications of laparoscopic antireflux surgery. AB - Over the last decade, the laparoscopic approach to antireflux surgery has been widely applied, resulting in improved early outcomes and greater patient acceptance of surgery for gastroesophageal reflux disease. However, although short-term outcomes are probably better overall than those following open surgery, it has become apparent that the laparoscopic approach is associated with an increased risk of some complications, and as well as the occurrence of new complications specific to the laparoscopic approach. Significant complications include acute paraesophageal hiatus herniation, severe dysphagia, pneumothorax, vascular injury, and perforation of the gastrointestinal tract. The incidence of some of these complications decreases as surgeons gain experience; others can be minimized by using an appropriate operative technique. In addition, laparoscopic reintervention is usually straightforward in the 1st postoperative week. For this reason, the surgeon should have a low threshold for early laparoscopic reexploration, facilitated by early radiological contrast studies, in order to reduce the likelihood that problems will arise later. PMID- 11395815 TI - The surgical management of congenital liver cysts. AB - BACKGROUND: Most series that report the results of surgical treatment for congenital liver cysts focus more on the technical aspects of the operation than on the late outcome of these patients. In this paper, we emphasize the importance of appropriate patient selection and adequate surgical technique for successful long-term outcome. METHODS: Twenty-four consecutive patients with congenital liver cysts were selected for surgical treatment. According to our own classification, 13 patients had simple liver cysts, nine had multicystic liver disease, and two had type I polycystic liver disease. All of these patients were treated by the fenestration technique. An open approach was used for five patients (group 1) treated between 1984 and 1990. In 19 patients (group 2) treated since 1991, a laparoscopic approach was used. The incidence of complicated liver cysts was 40% in group 1 and 68% in group 2. RESULTS: There were no treatment-related deaths in this series. The mean postoperative hospital stay was significantly shorter for patients who underwent successful laparoscopic fenestration (p < 0.05). In the open group (group 1), there were no postoperative complications, and all patients were alive and free of symptoms during a mean follow-up of 130 months, without any sign of cyst recurrence. In the laparoscopic group (group 2), four patients were converted to open surgery. One of these patients had an inaccessible posterior cyst; another had bile within the cystic cavity. A further two cases had complicated liver cysts with an uncertain diagnosis between congenital and neoplastic cysts. Four patients (21%) developed peri- or postoperative complications. During a mean follow-up time of 38.5 months, none of the patients with simple liver cysts incurred late symptoms or signs of cyst recurrence. In the six patients with multicystic liver disease, one developed disease-related cyst progression (17%) and required reoperation. One of the two patients with type I polycystic liver disease (50%) developed asymptomatic disease-related cyst progression. CONCLUSIONS: When patients are carefully selected and a proper surgical technique is employed, excellent long term results with a low morbidity rate can be achieved in patients with congenital liver cysts. Patients with multicystic liver disease or type I polycystic liver disease are more prone to late cyst recurrence. A tailored approach is thus indicated for patients with congenital liver cystic disease. However, the laparoscopic approach appears to be the gold standard for the treatment of highly symptomatic or complicated simple liver cysts. PMID- 11395816 TI - The role of laparoscopic ultrasound in the minimally invasive management of symptomatic hepatic cysts. AB - BACKGROUND: Now that the laparoscopic treatment of symptomatic liver cystic disease has proven feasible and safe, it is gaining wide acceptance. However, due to diagnostic pitfalls and a relatively high recurrence rate, further improvements and refinement of the procedure are still needed. We have evaluated the contribution of laparoscopic ultrasound in the diagnosis and management of patients with symptomatic liver cysts. METHODS: Twelve patients with single or multiple cysts of the liver and two patients with polycystic liver disease were managed laparoscopically. Laparoscopic ultrasound served as an integral part of the procedure in all patients. RESULTS: Patients underwent either complete cyst excision (two cases) or resection of the extrahepatic cystic component (eight cases). Additionally, in two patients, deep cysts not demonstrated by preoperative imaging studies were detected and treated with a combination of laparoscopy and laparoscopic ultrasound. In one patient with a cystobiliary fistula, conversion to an open cystjejunostomy was necessary. Patients with polycystic liver disease underwent a combination of excision and unroofing of both superficial and deeper cysts using laparoscopic contact ultrasound throughout the procedure. Laparoscopic ultrasonography was found to have a significant impact on the operative strategy in five patients (36%) with multiple cysts or polycystic disease. The postoperative course was uneventful in all cases. Thirteen patients remained asymptomatic throughout the follow-up period of 30 months; one patient with polycystic liver disease developed recurrent symptoms after 5 months and was treated with left hepatectomy. CONCLUSION: Additional use of laparoscopic ultrasound enables the detection, differentiation, and treatment of deep, nonvisualized cystic lesions (two patients, 16.6%) and validation of the adequacy of the laparoscopic procedure. PMID- 11395817 TI - Effects of amniodistention with carbon dioxide on fetal acid-base status during fetoscopic surgery in a sheep model. AB - BACKGROUND: Because the data from previous experiments on the fetal effects of carbon dioxide (CO2) amniodistention in endoscopic fetal surgery are conflicting, we set out to evaluate the fetal acid-base status during CO2 amniodistention, with or without maternal hyperventilation, using a sheep model for endoscopic surgery. METHODS: We assigned 26 pregnant ewes undergoing amniodistention with CO2 (4-5 mmHg intraamniotic pressure) to one of the following three groups: group I had fetal surgery + no maternal hyperventilation (n = 10); group II had fetal surgery + maternal hyperventilation (n = 10); group III had no fetal surgery + maternal hyperventilation (n = 6). Hyperventilation kept CO2 at 29-31 mmHg; in its absence, pCO2 ranged from 38 to 41. Fetal surgery consisted of fetoscopic tracheal clipping. Maternal blood pressure (mean, 98/69 mmHg) and heart rate (mean, 72 bpm) were kept at values comparable to human pregnancy. Fetal and maternal blood gas measurements were taken every 15 min during 1 h of amniodistention. RESULTS: The ranges for baseline mean fetal pCO2 (mmHg) and pH were 51-55 and 7.24-7.25, respectively, in all study groups. After 1 h of amniodistention, mean +/- SEM values of fetal pCO2 and pH were 88 +/- 3 and 7.06 +/- 0.03 in group I, 69 +/- 4 and 7.13 +/- 0.02 in group II, and 71 +/- 5 and 7.14 +/- 0.04 in group III, respectively. Therefore, maternal hyperventilation attenuated but could not prevent significant fetal hypercarbia and acidosis. Fetal surgical manipulation had no effect on these observations. CONCLUSION: CO2 amniodistention should not be considered for clinical practice until ways of preventing its effects on the fetal acid-base status can be demonstrated. PMID- 11395818 TI - The role of laparoscopy in the management of childhood intussusception. AB - BACKGROUND: Some authors have argued that intussusception is best treated via a laparoscopic approach. As we did not have this impression, we reviewed our experience with this condition. METHODS: : We reviewed all patients with intussusception who were treated at our hospital over the past 10 years. The choice of whether to use a laparoscopic or open approach depended on the patient's clinical condition and the availability of surgeons with laparoscopic expertise. RESULTS: A total of 72 patients were identified. Based on age, two subgroups were distinguished-one comprised of patients under the age of 3 years and one of patients over the age of 3 years. Sixty-five patients were under 3 years of age. Thirty-five had surgery, and 19 required resection. Of the 10 patients who were treated with a laparoscopic approach, only three could be reduced laparoscopically. After conversion in the other seven patients, the intussusception was reduced in five whereas a resection was required in two cases. Seven patients were 3 years of age or older. All of them underwent surgery, and all but one required resection. All four children who were laparoscoped subsequently had a bowel resection at open surgery. CONCLUSIONS: Patients 3 years of age or older usually need resection and will not benefit from the laparoscopic approach. Under 3 years of age, little is to be gained from a laparoscopic approach, provided good nonsurgical reduction facilities are available. There is a place for the laparoscopic approach in cases of recurrent intussusception or doubtful reduction. PMID- 11395819 TI - Is laparoscopic appendectomy safe in pregnant women? AB - BACKGROUND: Because of limited laboratory and clinical data, no accepted guidelines concerning the safety of laparoscopic appendectomy (LA) in pregnancy have been established yet. In this prospective study, we evaluated the safety and outcome of LA in pregnant women as compared with the same control group of pregnant women who underwent open appendectomy (OA) during the same period. METHODS: During the years 1996 to 1999, 11 consecutive pregnant women (mean age, 27 years; range 21-39 years; gestation age range, 7-34 weeks) who underwent LA were prospectively evaluated and compared with a matched group of 11 women (mean age, 30 years; range 18-42 years; gestation age range, 11-37 weeks) who underwent OA. The following parameters were analyzed: obstetric and gynecologic risk factors, length of procedure, perioperative complications, length of stay, and outcome of pregnancy. Both groups were well matched in age and risk factors for pregnancy loss. RESULTS: There was no significant difference in the length of procedure (60 vs. 46 min) and the complications rate (one in each group) between the LA and OA groups, respectively. There was no conversion in the LA group. The length of postoperative stay was shorter in the LA group (3.6 vs 5.2 days; p = 0.05). There was no fetal loss or other adverse outcome of pregnancy in either group, and all the women in both groups had normal full-term delivery. The infants' development was normal in both groups for a mean follow-up period of 30 months. CONCLUSIONS: According to this relatively small-scale study laparoscopic appendectomy in pregnant women may be as safe as open appendectomy. This procedure is technically feasible in all trimesters of pregnancy and associated with the same known benefits of laparoscopic surgery that nonpregnant patients experience. PMID- 11395820 TI - The impact of conventional and laparoscopic colon resection (CO2 or helium) on intraperitoneal adhesion formation in a rat peritonitis model. AB - BACKGROUND: Although postoperative intra-abdominal adhesion formation has been shown to be less with laparoscopic procedures than with open surgery, the extent of intraperitoneal adhesion formation after open and laparoscopic colon resection in patients with intra-abdominal infection remains unclear. METHODS: A standardized fecal inoculum was intraperitoneally applied in 72 rats to induce peritonitis. The rats were randomized into four groups. The three experimental groups underwent laparoscopic resection of the cecum with carbon dioxide (n = 20) or helium (n = 20) insufflation at a pressure of 8 mmHg, or conventional open cecum resection (n = 20). In the control group, no further manipulations were performed after induction of the peritonitis (n = 12). Blood samples were taken during the perioperative course to determine the plasma levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10). The number of intraperitoneal adhesions and the bacterial species of peritoneal swabs were determined in each animal. RESULTS: The adhesions were increased in all operation groups as compared with the control group. The number of adhesions was significantly greater after conventional resection than after laparoscopic approaches (p < 0.01). The overall adhesion score was significantly lower in the helium group (p < 0.05) than in the two other operation groups. There was no difference between the laparotomy and carbon dioxide groups. Whereas postoperative TNF-alpha plasma levels were decreased, IL-10 levels were significantly greater in the helium group than in the other three groups. CONCLUSIONS: Intraperitoneal infection is reducing the benefit of laparoscopic surgery regarding postoperative adhesions. Although laparoscopic resection showed an advantage in the number of adhesions with both gases, the total adhesion score was lowest in the helium group. PMID- 11395821 TI - Laparoscopic vs open appendectomy in overweight patients. AB - BACKGROUND: Laparoscopic appendectomy (LA) has been associated with a faster recovery and less postoperative pain than the open technique. However, few data are available on the clinical outcome of LA in overweight patients. METHODS: A group of 106 patients with a body mass index (BMI) > 26.4, representing the upper quintile of 500 prospectively randomized patients, were included in the study. They were randomized to undergo either laparoscopic or open appendectomy (OA). Operating and anesthesia times, postoperative pain, complications, hospital stay, functional index (1 week postoperatively), sick leave, and time to full recovery were documented. RESULTS: In OA, the operating time for overweight patients was significantly longer than that for patients in the normal weight range (40 vs 35 min, p < 0.05). In LA, there was no difference in operating time between the normal and overweight patients. Overweight patients who underwent LA had longer operating and anesthesia times than their OA counterparts (55 vs 40 min, p < 0.001; and 125 vs 100 min, p < 0.001, respectively). Postoperative pain was significantly greater in overweight patients who underwent OA than in those treated with the laparoscopic technique. Postoperative pain was also significantly greater in overweight patients subjected to OA than in patients of normal weight after 4 weeks; the clinical significance may, however, be of less importance since the values are low (0.26 vs 0.09, p < 0.05). There were no significant differences between the two operating techniques in terms of complications. Hospital stay was longer for overweight patients than for normal weight patients undergoing OA (3.0 vs 2.0, p < 0.01). The functional index did not differ between any group of patients. Sick leave was longer for overweight patients who underwent OA than for normal-weight patients treated with the same technique (17 vs 13 days, p < 0.01). In the laparoscopic group, however, there were no differences between the overweight and normal-weight patients. Time to full recovery was greater in overweight patients subjected to OA than in the overweight patients in the LA group (22 vs 15 days, p < 0.001). CONCLUSION: In this study, overweight patients who were submitted to LA had less postoperative pain and a faster postoperative recovery than overweight patients who had OA. LA also abolished some of the negative effects that overweight had on operating time, hospital stay, and sick leave with the open technique. However, anesthesia and operating times were significantly longer in LA for both overweight patients and those with a normal BMI. PMID- 11395822 TI - Laparoscopic gastric bypass as a reoperative bariatric surgery for failed open restrictive procedures. AB - An estimated 2% of men and 6% of women in the United States are morbidly obese. These patients have a mortality rate that is six to twelve times greater than their normal-weight peers. In these extreme cases of overweight, only surgical intervention can produce substantial and sustained weight loss. However, up to 20% of these patients will require reoperation for failure to achieve or maintain an adequate weight loss. Reoperative bariatric surgery can be challenging and has traditionally been performed as an open procedure. We present seven female patients who underwent a laparoscopic Roux-en-Y gastric bypass after failing to achieve their weight-loss goals with prior open restrictive procedures. Their average BMI before surgical revision was 42.2 kg/m2. Mean operative time was 4 h 20 min, and length of hospital stay was 4.1 days. There were three complications in two patients and no deaths. Our initial experience suggests that laparoscopic gastric bypass is a safe and feasible reoperative bariatric procedure. Confirmation awaits a larger series of patients with follow-up data. PMID- 11395823 TI - Treatment of acute cholecystitis. A comparison of open vs laparoscopic cholecystectomy. AB - BACKGROUND: In this study, the clinical results and cost-effectiveness of open vs laparoscopic cholecystectomy in the treatment of acute cholecystitis were compared. METHODS: Over a 5-year period (1994-98), 894 cholecystectomies were performed, 545 (60.96%) of them laparoscopically and 349 (39.04%) by the open method. The study included 209 patients with a clinical diagnosis of acute cholecystitis; 115 (55.02%) of them were operated on by the open method and 94 (44.98%) by the laparoscopic method. RESULTS: A comparison analysis revealed that the mean postoperative treatment period was 8.40 days after open and 4.38 days after laparoscopic cholecystectomy. In the group operated on by the open method, 106 patients received an antibiotic, a mean of 5.09 ampules and 3.2 tablets or suppositories of an analgesic, and 2.91 dressings per patient; whereas in the group submitted to the laparoscopic method, the comparable figures were 43, 3.13, 2.1, and 1.47, respectively. In 31 (26.96%) employed patients operated on by the open method, the mean absenteeism from work was 42 days; whereas in 31 (32.98%) of those operated on by the laparoscopic method, it was 17 days. The mean operating times for the procedures were 89 and 115 min for the open and laparoscopic methods, respectively. Two patients submitted to open cholecystectomy died within 30 days postoperatively. Wound infection was recorded in 10 (8.7%), prolonged biliary secretion in two, and cicatricial hernia in five (4.35%) patients. In the group submitted to laparoscopic cholecystectomy, there were no deaths; nine (9.57%) conversions were required; four patients had to be reoperated on, two of them for bile lobe hemorrhage and two for massive biliary secretion from the open cystic duct; herniation at the site of supraumbilical incision developed in three patients, and infection developed at the same site in two (2.13%) patients. The hospital cost was significantly higher in laparoscopic patients ($1181 vs $873) USD), as was the total cost of treatment for acute cholecystitis ($1430 vs $1316). However, the cost for sick leave and rehabilitation was significantly lower in laparoscopically treated patients ($486 vs $1199). CONCLUSIONS: Our comparison analysis of the results and cost effectiveness of the surgical treatment of acute cholecystitis clearly pointed to the advantages of laparoscopic over open cholecystectomy-i.e., better clinical outcome and a more rapid resumption of daily activities. Hospital and total costs of treatment were on average higher in laparoscopic patients, except for the employed ones, where the lower sick leave cost translated into a significant reduction in total costs. PMID- 11395824 TI - Clinical efficacy of video-assisted gasless transanal endoscopic microsurgery (TEM) for rectal carcinoid tumor. AB - BACKGROUND: We used video-assisted gasless transanal endoscopic microsurgery (TEM) to evaluate 12 consecutive patients for carcinoid tumor with pathologically typical features in the proximal rectum. The study parameters included feasibility, results and final outcome. METHODS: A scope with an optical angle of 25 and a 7-mm diameter was used for TEM. The carcinoid tumor was resected in full thickness. Primary closure was accomplished using a running suture of 3-0 braided absorbable suture. RESULTS: The tumor in these patients was <15 mm in diameter. It was localized within the submucosal layer on the oral side from the peritoneal reflection and unreachable via Park's transanal approach. The mean duration of the operation was 63 min. The postoperative course in all patients was free from significant complications, and all patients were discharged within 1 week. None of the patients had postoperative pain or required sedatives. CONCLUSION: Video assisted gasless TEM is a simple and minimally invasive procedure to treat benign carcinoid tumors in the proximal rectum. PMID- 11395825 TI - Effect of insufflation gas and intraabdominal pressure on portal venous flow during pneumoperitoneum in the rat. AB - BACKGROUND: Carbon dioxide, the primary gas used to establish a pneumoperitoneum, causes numerous systemic effects related to cardiovascular function and acid-base balance. Therefore, the use of other gases, such as helium, has been proposed. Furthermore, the pneumoperitoneum itself, with the concomitant elevation of intraabdominal pressure, causes local and systemic effects that have been only partly elucidated. Portal blood flow, which plays an important role in hepatic function and cell-conveyed immune response, is one of the affected parameters. METHODS: An established animal model (rat) of laparoscopic surgery was extended by implanting a periportal flow probe. Hemodynamics in the portal vein were then measured by transit-time ultrasonic flowmetry during increasing intraabdominal pressure (2-12 mmHg) caused by gas insufflation (carbon dioxide vs helium). RESULTS: The installation of the pneumoperitoneum with increasing intraperitoneal pressure led to a significant linear decrease in portal venous flow for both carbon dioxide and helium. At higher pressure levels (8-12 mmHg), portal blood flow was significantly lower (1.5-2.5-fold) during carbon dioxide pneumoperitoneum. An intraabdominal pressure of 8 mmHg caused a decrease to 38.2% of the initial flow (helium, 59.7%); whereas at 12 mmHg, portal flow was decreased to 16% (helium, 40.5%). CONCLUSION: Elevated intraabdominal pressure generated by the pneumoperitoneum results in a reduction of portal venous flow. This effect is significantly stronger during carbon dioxide insufflation. Portal flow reduction may compromise hepatic function and cell-conveyed immune response during laparoscopic surgery. PMID- 11395826 TI - Minimally invasive video-assisted parathyroidectomy. AB - BACKGROUND: The standard surgical procedure for parathyroidectomy consists of bilateral cervical exploration and the visualization of all four parathyroid glands. However, improved preoperative localization techniques and the availability of intraoperative intact parathyroid hormone (iPTH) monitoring now allow single adenomas to be treated with minimally invasive techniques. METHODS: Patients with primary hyperthyroidism (pHPT), who were found to have one unequivocally enlarged parathyroid gland on preoperative ultrasound and 99mTc SestaMIBI scintigraphy underwent minimally invasive video-assisted parathyroidectomy by an anterior approach. Intraoperatively, rapid electrochemiluminescense immunoassay was used to measure iPTH levels shortly before and 5, 10, and 15 mins after excision of the adenoma. The operation was considered successful when a >50% decrease in preexcision iPTH levels was observed after 5 min. RESULTS: Between November 1999 and May 2000, 10 of 22 patients with pHPT were deemed eligible for the minimally invasive approach. In all cases, the adenoma was removed successfully. However, in two cases, intraoperative iPTH monitoring did not show a sufficient decrease in iPTH values. Subsequent cervical exploration revealed a double adenoma in one case and hyperplasia in the other. CONCLUSIONS: Even when high-resolution ultrasound and 99mTc-SestaMIBI scintigraphy are used, the presence of multiple glandular desease cannot be ruled out entirely. When the minimally invasive approach is contemplated, intraoperative iPTH monitoring is indispensible to ensure operative success. However, in selected cases, minimally invasive parathyroidectomy represents an excellent alternative to the conventional technique. PMID- 11395827 TI - Laparoscopic splenopexy by peritoneal and omental pouch construction for intermittent splenic torsion ("wandering spleen"). AB - Wandering spleen is an extremely rare anatomic variant with potentially serious clinical implications. Usually, splenectomy is advocated for treatment of this disease. Various methods for preserving the wandering spleen by means of splenopexy have been described, including two reports on laparoscopic splenic refixation. We describe the third case in which laparoscopic splenopexy was used to manage chronic intermittent splenic torsion. In a 25-year-old woman, splenopexy was successfully performed by laparoscopic reposition and fixation of the spleen by omental pouch creation. At laparoscopy with a normal operating room setup and four trocars, a free-floating, macroscopically normal spleen attached to an abnormally long vascular pedicle with no gastrosplenic or phrenicosplenic ligaments was detected in the lower right quadrant. The spleen was repositioned and placed in the left phrenorenal angle. Splenopexy was achieved by suturing the left colophrenic ligament to the lateral diaphragm, thus creating a pouch for the inferior part of the spleen, and by suturing the gastrocolic ligament to the anterior diaphragm to create a pouch for the upper splenic pole. The postoperative course was uneventful. At a follow-up examination 3 months after the operation, the patient was well, with no further episode of recurrent abdominal pain. Ultrasonographically, the spleen was seen easily in the left hypochondrium in its normal physiologic position. Laparoscopic splenopexy is a useful option for organ-preserving therapy of the wandering spleen. PMID- 11395828 TI - Acute hemoperitoneum caused by rupture of omentum adhesions after running. AB - The case of a 37-year-old man in whom a massive hemoperitoneum developed a few hours after running is described. The patient disclaimed any trauma and clearly noted that symptoms appeared after running. Findings at laparoscopy showed that the bleeding was caused by the rupture of adhesions between the omentum and left inguinal abdominal wall. These adhesions, which had resulted from a previous laparoscopic transperitoneal bilateral inguinal hernia cure, were resected. Recovery was simple and follow-up assessment was uneventful. Hemoperitoneum secondary to the rupture of intraperitoneal adhesions is very rare in the absence of precipitating trauma. However, the trauma can be trivial. Rupture of intra abdominal adhesions has been described after sexual intercourse or mobilization of the patient under general anesthesia. Disruption of adhesions by insufflation or mobilization of organs under laparoscopy also is reported. The transperitoneal approach to laparoscopic treatment of inguinal hernia can be responsible for late intestinal obstruction caused by intra-abdominal adhesions, but late hemorrhagic complication has not yet been reported. PMID- 11395829 TI - Splenosis in a port site after laparoscopic splenectomy. AB - Splenosis, the autotransplantation of splenic tissue, is most commonly seen after traumatic splenic rupture and splenectomy. It also can occur during embryonic development. Intraperitoneal, intrathoracic, and retroperitoneal sites have been reported. Although the presence of the splenic tissue often is asymptomatic and an incidental finding, it may present with pain or be confused with various pathologies including neoplasia. Because most pediatric splenectomies are performed for hemolytic disorders, parenchymal disruption must be contained to avoid recurrent disease. We present a case in which the devascularized spleen was contained in a bag and fragmented in situ. Splenosis developed in the retrieval port site after laparoscopic splenectomy and cholecystectomy. Port-site splenosis needs to be considered in the differential diagnosis of port-site pain and a palpable nodule postsplenectomy. PMID- 11395830 TI - Stenting...where credit is due. PMID- 11395831 TI - Radially dilating trocars are not cost-effective alternatives except for pediatric laparoscopic operations, especially those for undescended testis and imperforate anus. PMID- 11395832 TI - The "lap-loop" as a substitute for the fourth trocar in laparoscopic cholecystectomy. A critical appraisal. PMID- 11395834 TI - ASPAN's research vision. PMID- 11395835 TI - Is meta-analysis clinically useful for perianesthesia nurses? AB - Meta-analyses can provide clinicians with the evidence-based information necessary to effect change in patient care delivery. Meta-analysis, the application of statistical techniques to condense data from a group of individual studies, is the most sophisticated summary of research evidence. An example meta analysis study of inadvertent surgical hypothermia is used in this article to describe how nurses can locate, evaluate, and apply synthesized research data for perianesthesia clinical practice. Without data to support decisions, nurses will increasingly be faced with decisions made by others based on cost-cutting measures alone. Although meta-analysis may resolve a controversy or solve a clinical problem, it will not provide simple statistical answers for complex problems or obviate the need for sound and compassionate clinical judgment. PMID- 11395836 TI - The effects of acupressure on the incidence of postoperative nausea and vomiting in postsurgical patients. AB - Postoperative nausea and vomiting (PONV) is one of the most common postoperative complications. Aside from pharmacological interventions, other complementary healing modalities have been introduced to assist patients in decreasing PONV and improving postoperative outcomes. This study examined acupressure as a potentially holistic and safe complement to the more traditional approach of using drugs to prevent and/or relieve nausea and vomiting in the postoperative patient. Acupressure involves constant pressure (without puncture of the skin) on the Nei Guan acupuncture points through the use of a British product called Sea Bands (Sea Band UK Ltd, Leics, England). These bands are made of elasticated fabric, with a small round plastic button inside. A quasi-experimental research design was used to examine the effects of unilateral and bilateral application of acupressure on 157 patients who are prone to PONV: postgynecological, postplastic, and posturological surgery patients. The incidence of PONV was determined through retrospective chart reviews. The hypothesis was that there would be a difference in the incidence of PONV between 5 groups: group 1 (Sea Bands with acupressure on both wrists), group 2 (Sea-Bands with acupressure on one wrist), group 3 (wristband without acupressure on both wrists), group 4 (wristband without acupressure on one wrist), and group 5 (no wristband). This hypothesis was examined by using a one-way analysis of variance (ANOVA); it was not supported. Neither unilateral nor bilateral application of acupressure significantly affected the incidence of nausea and vomiting. These findings must be viewed with caution, however, because power analysis showed low effect sizes and an inadequate sample size. Further research is recommended with a larger sample size. This study has made perianesthesia nurses more aware of other complementary modalities to assist patients with nausea and vomiting. PMID- 11395837 TI - A study of perianesthesia nursing practice: the foundation for newly revised CPAN and CAPA certification examinations. AB - The CPAN and CAPA nursing certification programs, sponsored by ABPANC, are designed to promote and enhance the quality of care delivered to patients who have received anesthesia. This article describes ABPANC's most recent Role Delineation Study (RDS), also called the Study of Practice. The work of a think tank that convened to reconceptualize the organizing framework for the certification examination programs is described, including the results of an environmental analysis. A variety of methods were used to conduct the study. As a result of study findings, the CPAN and CAPA programs have been redesigned to focus on the physiological, behavioral and cognitive, safety, and advocacy needs of perianesthesia patients. Those needs and the knowledge base that is required of perianesthesia nurses to meet those needs are defined. The first CPAN and CAPA examinations using the new framework will be given November 3, 2001. PMID- 11395838 TI - Evaluation of ASPAN's preoperative patient teaching videos on general, regional, and minimum alveolar concentration/conscious sedation anesthesia. AB - This descriptive study was undertaken as part of a clinical improvement effort by the ASPAN Research and Education Committees to evaluate adult patients' perception of and satisfaction with the ASPAN Preoperative Patient Teaching videotape on general, regional, and minimum alveolar concentration (MAC)/conscious sedation anesthesia. Research findings on the use of videotapes for preoperative education are mixed. Some studies have reported that the use of videotapes increases knowledge and decreases anxiety, whereas other studies have shown a minimal effect on knowledge and anxiety. A convenience sample of 96 adult patients was chosen from those who were scheduled for surgeries with the above anesthesia techniques in 11 US hospitals and/or surgical centers within 4 ASPAN regional boundaries. Patients viewed the videotape the day(s) before surgery and then completed ASPAN's Preoperative Anesthesia Patient Teaching Questionnaire to measure patient perception and satisfaction. Sixty percent of the patients were women, and 50% had a college degree or higher. The average age of the patients was 51 (+/-17.2). Overall satisfaction scores had a potential range of 10 to 40, with higher scores indicating greater satisfaction. The mean satisfaction score for this study was 35 (+/-6.6). No significant relationships were found between satisfaction with the videotape and age, gender, or educational level. Patients were asked to rank each of 4 teaching methods. Among the choices of individualized instruction, written materials, Internet-based instruction, and videotape, the videotape method was ranked as most preferred. The information obtained from this study will be used to modify and improve the content of the patient education videotape produced by ASPAN. PMID- 11395839 TI - Effects of hypoxia on diaphragm activity in anesthetized rats. AB - Hypoxia is one of the most common conditions observed by PACU nurses after surgery. It may be caused by a reduced functional residual capacity, hypoventilation, and/or ventilation-perfusion mismatch. Hypoxia can also affect diaphragm contractility, making it difficult to wean postoperative patients from mechanical ventilation. Clinically, however, there is no method to directly measure diaphragm contractility; therefore, indicators of intrathoracic pressure such as tidal volume are used. The purpose of this study was to directly measure the effects of diaphragm shortening in 12 anesthetized Sprague-Dawley rats before, during, and after induced hypoxia. A miniaturized ultrasonic sensor was used to measure changes in diaphragm thickness as an index of diaphragm shortening. A stainless steel electrode was attached adjacent to the ultrasonic sensor and used to measure the electromyogram (EMG) of the diaphragm. After normoxic measurements were recorded, hypoxia was initiated by connecting the tracheal cannula to a latex balloon containing 7.4% oxygen in nitrogen. During the first 5 minutes of hypoxia, diaphragm shortening, EMG, and intrathoracic pressure increased. Over the next 30 to 100 minutes, EMG and intrathoracic pressure remained elevated, whereas diaphragm shortening decreased to 50% of control, which was defined as diaphragm fatigue. The mean time for hypoxia induced diaphragm fatigue to occur was 63 minutes. These results indicate that hypoxia-induced decline in diaphragm shortening was not caused by a decrease in muscle excitation as measured by EMG. These data suggest that impairment in mechanical-chemical coupling (diaphragm shortening) could be a result of decreased oxygen availability associated with the lower arterial blood oxygen content. Thus, the increase in intrathoracic pressure throughout hypoxia suggests that intrathoracic pressure is not always a consistent index of the contractile state of the diaphragm. PMID- 11395840 TI - A comparison study on the effects of prewarming patients in the outpatient surgery setting. AB - Maintenance of core body temperature in surgical patients presents a challenge to perioperative nurses. Core temperatures less than 36 degrees C are associated with multiple adverse outcomes postoperatively. Internal redistribution of heat from the body core to the colder periphery results in core temperature decreases of 0.5 degrees C to 1.5 degrees C in the first 30 minutes after induction of anesthesia. The purpose of this study was to determine if there was a difference in arrival temperatures to the PACU between surgical patients who had been warmed preoperatively with a forced warm air blanket and those patients warmed with cotton blankets. One hundred patients were randomly assigned to receive prewarming by using a forced-air warm blanket (n = 50) or a cotton blanket (n = 50). Temperatures were monitored every 15 minutes throughout the preoperative and postoperative periods. Patients in the forced warm air group had significantly higher temperatures on arrival to the PACU from the OR than did patients in the warm blanket group (P =.000). Patients in the forced warm air group exhibited a change in temperature of 0.0067 degrees C (+/-.52) compared with a decrease of 0.22 degrees C (+/-.48) for patients in the control group. PMID- 11395841 TI - Performance assessment and improvement. PMID- 11395842 TI - A successful practice: pre-admitting test center. PMID- 11395843 TI - The American Society of Anesthesiologists addresses herbal medications. PMID- 11395844 TI - Just curious. PMID- 11395845 TI - Biological basis of anemia. AB - Anemia is a frequent complication in cancer, occurring in more than 50% of patients with malignancies. Several factors can cause anemia in these patients, such as blood loss, hemolysis, bone marrow infiltration, hypersplenism, and nutrient deficiencies. However, in a considerable number of patients, no cause other than malignant disease itself can be implicated. This cancer-related anemia is similar to the anemia observed in other chronic diseases, such as rheumatoid arthritis and some chronic infections. The syndrome of anemia of chronic disease is characterized by a hyporegenerative, normocytic, normochromic anemia associated with reduced serum iron and transferrin saturation but elevated (or normal) ferritin levels. Cancer-related anemia results from activation of the immune and inflammatory systems, leading to increased release of tumor necrosis factor, interferon-gamma, and interleukin-1. The cytokine-mediated relative failure of erythropoiesis has been further investigated, and three different mechanisms of action are proposed: (1) impaired iron utilization; (2) suppression of erythroid progenitor cells differentiation; and (3) inadequate erythropoietin production. In addition, the life span of red blood cells is shortened in cancer related anemia and production cannot compensate sufficiently for the shorter survival time. Administration of recombinant human erythropoietin (r-HuEPO, epoetin alfa) can not only correct inadequate endogenous erythropoietin production, but also can overcome the suppression of erythroid progenitor cells and impairment of iron mobilization. PMID- 11395846 TI - Symptomatology of anemia. AB - The symptoms and severity of anemia depend on various factors, including the degree of anemia, the rapidity of its onset, and the age and physiologic status of the patient. Although the human body tries to counterbalance the effects of anemia by various mechanisms, almost every organ system of the human body is eventually affected. The symptoms experienced by patients vary from cold skin, dizziness, and palpitations to pulmonary edema, heart failure, depression, and severe impairment of cognitive function. Anemia substantially impacts patients' quality of life, a fact that has been shown in several clinical trials in patients with renal disease as well as in patients suffering from various malignancies undergoing chemotherapy. These studies evaluated the administration of recombinant human erythropoietin (r-HuEPO, epoetin alfa) to anemic patients, and it was shown that raising hemoglobin levels with epoetin alfa ameliorated the symptoms of anemia and significantly improved the functional status and overall quality of life in cancer patients. Furthermore, preliminary data indicate that the correction of anemia in cancer patients may in addition improve treatment efficacy and possibly overall survival. PMID- 11395847 TI - Fatigue: a main component of anemia symptomatology. AB - Fatigue is a common complaint of patients with cancer or other physical or mental disorders. In cancer patients, the estimated prevalence of fatigue is high (about 70% to 90% in different surveys). However, despite the high prevalence and widely recognized clinical relevance of fatigue, few studies have been performed to evaluate the putative causal factors and therapeutic approaches for this condition. The paucity of studies has been mainly because of the lack of proper instruments to quantify this clinical problem. Moreover, fatigue is multifactorial, which makes evaluation of precise relationships with other medical conditions difficult. In particular, fatigue is considered the cardinal symptom of anemia. The pathogenesis of anemia-related fatigue remains unclear, but some suggest that abnormalities in energy metabolism play a role in inducing fatigue. In cancer patients, this effect may be exacerbated by the increased metabolic needs associated with tumor growth. At the clinical level, the relationship between anemia and fatigue is universally accepted. However, early studies were unable to show a clear association between fatigue and hemoglobin levels. Recently, new insights were afforded by the implementation of innovative survey instruments that assess the effects of fatigue and other (nonfatigue) symptoms of anemia on the patient's well-being and quality of life. The use of these validated instruments has shown a direct effect of hemoglobin levels on fatigue and other quality of life parameters. Thus, amelioration of anemia and fatigue should be considered a primary endpoint of antineoplastic and supportive care treatment of cancer patients. Accordingly, the search for new simplified methods of assessment of fatigue and other anemia-related symptoms and their treatment outcomes should be strongly encouraged. PMID- 11395848 TI - The erythropoietin receptor. AB - Erythropoietin (EPO) is the primary regulator of erythropoiesis, and promotes the survival, proliferation, and differentiation of erythroid progenitor cells. The EPO receptor belongs to the same family of receptors as growth hormone, granulocyte-colony stimulating factor, granulocyte macrophage-colony stimulating factor, and some interleukins. In the erythropoietic process, EPO induces homodimerization of the EPO receptor, which is located on the surface of erythroid progenitor cells. Dimerization activates the receptor-associated Janus kinase 2 via transphosphorylation. Specific tyrosines in the intracellular portion of the receptor are phosphorylated and serve as a docking site for intracellular proteins, including one of the signal transducers and activators of transcription (STAT5). This results in activating various cascades of signal transduction. STAT5 enters the nucleus on phosphorylation, inducing the transcription of erythroid genes. Phosphatases dephosphorylate Janus kinase 2 and downregulate the EPO receptor. Erythropoietin receptor activation seems to exert its effect by inhibiting apoptosis rather than by affecting the commitment of erythroid lineage, although the mechanism by which this occurs is as yet unclear. Anemia in cancer is associated with excessive production of cytokines that inhibit EPO synthesis, thereby interfering with the normal erythropoietic process, which leads to a reduction in red blood cells and the ability to oxygenate tissue. PMID- 11395849 TI - Erythrocytes and the transfer of anticancer drugs and metabolites: a possible relationship with therapeutic outcome. AB - Blood functions as a mobile tissue in an exchange system, with the remaining body tissue as a stationary phase. The equilibrium among plasma water, plasma proteins, and blood cells is described by models, but little consideration has been given to the substance-binding capacity of erythrocytes. There are numerous reasons for this, including bioanalytical limitations (ie, it has been difficult to study erythrocytes in the laboratory in their natural state). Erythrocyte monitoring requires accurate blood sampling and quantitative isolation of erythrocytes without disturbing the equilibrium of substances of interest between erythrocytes and plasma or other blood constituents. This became possible with the advent of the measurement of sediment device. The mass of a given substance available in blood can be described by M(Blood) = M(Plasma) + M(ERY) (+ M(REM)). M(ERY) is the mass of a substance present in erythrocytes and it is shown that for several oxazaphosphorines, such as iphosphoramide mustard, that M(ERY) determines M(Blood) with great superiority over M(Plasma). The impact of erythrocyte monitoring on therapeutic outcome has to be defined, but is an important area of research. PMID- 11395850 TI - Oxygen status of malignant tumors: pathogenesis of hypoxia and significance for tumor therapy. AB - Hypoxic areas are a characteristic property of solid tumors. Hypoxia results from an imbalance between the supply and consumption of oxygen. Major pathogenetic mechanisms for the emergence of hypoxia are (1) structural and functional abnormalities in the tumor microvasculature; (2) an increase in diffusion distances; and (3) tumor- or therapy-associated anemia leading to a reduced O2 transport capacity of the blood. There is pronounced intertumor variability in the extent of hypoxia, which is independent of clinical size, stage, histopathologic type, and grade. Local recurrences have a higher hypoxic fraction than primary tumors. Tumor hypoxia is intensified in anemic patients, especially in tumors with low perfusion rates. Tumor hypoxia is a therapeutic problem, as it makes solid tumors resistant to sparsely ionizing radiation and some forms of chemotherapy. Hypoxia also may modulate the proliferation and cell cycle position of tumor cells and, in turn, the amount of cells destroyed following therapy. Recent clinical studies suggest that hypoxia can enhance malignant progression and increase aggressiveness through clonal selection and genome changes. As a result, loss of differentiation and apoptosis, chaotic angiogenesis, increased locoregional spread, and enhanced metastasis can further increase resistance to therapy and affect long-term prognosis. Hypoxia is a powerful, independent prognostic factor in cervix cancers, carcinomas of the head and neck, and in soft tissue sarcomas. PMID- 11395851 TI - Biological consequences of tumor hypoxia. AB - Growing evidence from experimental and clinical studies points to the fundamental, pathophysiologic role of hypoxia in solid tumors. Intratumoral hypoxia is a consequence of a structurally and functionally disturbed microcirculation, with deterioration of the diffusion geometry and of tumor associated anemia. Hypoxia-induced changes of the proteome in the neoplastic and stroma cells may lead to neoplastic growth impairment through molecular mechanisms, resulting in cellular quiescence, differentiation, and apoptosis. Alternatively, hypoxia-induced proteome changes activating nonspecific stress response, anaerobic metabolism, angiogenesis, tissue remodeling, and change of cell contacts may promote tumor propagation by enabling neoplastic cells to overcome the nutritive deprivation through adaptation or escape from the "hostile" environment. Whether the phenotypic result of hypoxia-induced proteome change is impairment, stasis, or promotion of neoplastic growth is thought to be determined by the genome of the tumor cells and additional microenvironmental factors. Tumor cells with genomic alterations (such as loss of apoptotic potential) allowing their survival under hypoxia will aggravate tumor hypoxia. Sustained hypoxia increases genomic instability, genomic heterogeneity, and the selection pressure of the microenvironment. New variants even better adapted to survive and proliferate under reduced pO2 will be selected through clonal expansion. These variants will further drive the vicious circle of malignant progression which is clinically characterized by an increasing probability of local, perifocal, regional, and distant spread. PMID- 11395852 TI - The use of epoetin alfa to increase and maintain hemoglobin levels during radiotherapy. AB - Acute or chronic anemia, a common complication in cancer patients, is associated with the development of tumor hypoxia. It has been shown in several trials that decreased hemoglobin (Hb) levels inducing tumor hypoxia adversely impact radiotherapy or combined radiochemotherapy outcome. For example, pre- and post treatment Hb levels can be predictive factors for the outcome of radiotherapy, locoregional tumor control, and survival. Two strategies have been established to correct cancer-related anemia and improve radiotherapy outcome: immediate increase of Hb levels with transfusions, or treatment with recombinant human erythropoietin (r-HuEPO, epoetin alfa), which slowly but steadily increases Hb levels to within normal range. As transfusions are associated with severe risks and adverse events, the use of epoetin alfa to treat pre-existing anemia or prevent therapy-induced anemia represents an attractive strategy. The ability of epoetin alfa to maintain or to increase Hb levels in patients undergoing radiotherapy or radiochemotherapy have been shown in several studies in different tumor types. In addition to improving the results of radiotherapy and radiochemotherapy, anemia intervention with epoetin alfa may impact overall survival. PMID- 11395853 TI - The impact of hemoglobin levels on treatment outcomes in patients with cancer. AB - Clinical trials and surveys have shown that a majority of patients with cancer have low hemoglobin levels as a result of the disease and/or treatment. Clinical trials also have shown that the impact of anemia may be more insidious and far reaching than generally appreciated. Specifically, studies have shown that low hemoglobin levels have significant impact on treatment outcomes, including survival. The mechanisms by which treatment efficacy and survival are compromised have not been fully elucidated but may include cellular compromise (eg, impaired tumor oxygenation), or more general patient compromise (eg, decreased quality of life and treatment delivery). Recent studies have suggested that increasing hemoglobin levels with recombinant human erythropoietin (r-HuEPO, epoetin alfa) have resulted in better outcomes following radiotherapy, chemotherapy, and the combined-treatment modality. PMID- 11395854 TI - Prevalence of anemia in cancer patients undergoing radiation therapy. AB - Anemia is associated with reduced local tumor control and impaired quality of life in patients with several types of solid tumors. The prevalence of anemia in patients who present at radiation oncology departments has not been well documented, and the impact of anemia on the outcome of radiation therapy is not widely appreciated in the radiation oncology setting. In an ongoing study, we are retrospectively reviewing the medical charts of patients before and after radiation therapy at our institutions to determine the magnitude of the anemia problem in this population. Preliminary data are available for 574 randomly selected patients (52% female) seen between December 1996 and June 1999. At presentation, 41% of all patients were anemic (hemoglobin < 12 g/dL); by the end of radiation therapy, this percentage increased to 54%. The most common tumor types were prostate (16%), breast (14%), head and neck (12%), colorectal (11%), lung/bronchus (11%), and uterine-cervix (9%). Anemia was most prevalent in patients with uterine-cervical tumors (75%), increasing to 79% by the end of radiation therapy. The prevalence of lung/bronchus and colorectal cancer was 55% and 44%, respectively, at baseline and increased to 77% and 63%, respectively, after radiation therapy. For nearly all tumor types, the majority of patients had or developed mild to moderate anemia (hemoglobin 10.0 to 11.9 g/dL). These data show that anemia is widespread among patients seen in radiation oncology practices. However, the anemia is usually mild and readily correctable. Because anemia and hypoxia possibly associated with anemia are obstacles to local tumor control and maintenance of quality of life, strategies to reverse anemia should receive greater attention. PMID- 11395855 TI - The effect of hemoglobin level on radiotherapy outcomes: the Canadian experience. AB - Studies have shown that hemoglobin (Hb) level is a prognostic factor for outcomes following definitive radiotherapy in patients with a variety of tumors. We conducted a retrospective study on patients with carcinoma of the cervix to evaluate the relationships among Hb level, response to radiation, and effect on local control and survival. Data from 3 years (1989, 1990, 1992) were collected at seven institutions across Canada from women who received definitive radiotherapy, > or = 3,500 cGy, for carcinoma of the uterine cervix. The collected data included Hb levels at presentation and during radiation treatment specific for the time course of anemia and blood transfusion practice, patient demographics, details of the treatment regimen, and outcomes. Of 630 patients reviewed, 605 were eligible and were included in the analyses; median follow-up was 41 months. Twenty-five percent of patients received blood transfusions, most frequently when Hb decreased to below 100 g/L. Baseline Hb > or = 120 g/L was shown on univariate analysis to be significant for higher rates of pelvic disease control and disease-free survival. On multivariate analysis, baseline Hb was not a significant prognostic factor, but average weekly nadir Hb (AWNH) levels during radiotherapy were shown to be significant, and second only in importance to tumor stage. Five-year survival rates were similar if AWNH levels were > or = 12 g/L, regardless of whether baseline Hb was lower than 120 g/L (70%) or > or = 120 g/L (74%); the difference in 5-year survival between patients with lower AWNH (< 120 g/L) and higher AWNH (> or = 120 g/L) was significant. Patients with higher AWNH had significantly lower rates of overall relapse, local recurrence, and distant metastases. A stepwise significant increase was noted in overall survival rate as AWNH levels increased, regardless of transfusion status. Hemoglobin levels > or = 120 g/L during radiotherapy were shown to be a significant prognostic factor for successful radiotherapy and disease-free survival. There was no difference in outcomes relative to transfusion status. PMID- 11395857 TI - At the crossroad of conception and infection: initiatives for immunoglobulin based contraceptive R & D. AB - Better understanding of the immunological mechanisms implying the insemination and the infertility of some men and women is needed and crucial to the development of an effective immunocontraceptive method. To provide good protection against conception or infection, and avoid any possible and unexpected complications which immunocontraceptive "vaccine" may arise of, it seems the right time for scientists to create a virtually new thinking for this extremely urgent and important issue. This conceptual article describes our original thoughts of the future development of immunocontraceptives, preferably, based on immunoglobulins rather than vaccines, against human sperm specific antigens and seminal plasma immunosuppressive factors. Its general correctness, advantages and feasibility for fertility regulation and prevention of infection are discussed. PMID- 11395856 TI - Effects of epoetin alfa on the central nervous system. AB - Erythropoietin (EPO) is a glycoprotein that has been shown to mediate response to hypoxia, and is most notably recognized for its central role in erythropoiesis. In a series of experiments using rodent models, the ability of systemically administered recombinant human erythropoietin (r-HuEPO, epoetin alfa) to cross the blood-brain barrier and affect the outcome of neuronal injury or cognitive function was evaluated. It was shown that EPO and EPO receptors are expressed at capillaries of the brain-periphery interface, and that systemically administered epoetin alfa crossed the blood-brain barrier. Compared with control animals, epoetin alfa significantly reduced tissue damage in an ischemic stroke model when administered 24 hours before inducing stroke, with significant protection still evident when epoetin alfa was administered 6 hours poststroke. Epoetin alfa reduced injury by blunt trauma when administered 24 hours before trauma, with a significantly smaller volume of tissue necrosis noted when compared with controls. The observation that epoetin alfa may reduce nervous system inflammation was confirmed when an experimental autoimmune encephalomyelitis model in which rats were shown to have significantly delayed onset and reduced severity of experimental autoimmune encephalomyelitis symptoms after treatment with epoetin alfa. Epoetin alfa also was shown to ameliorate the latency and severity of seizures, and significantly increase survival versus controls when exposed to kainate. These findings suggest future potential therapeutic uses for epoetin alfa beyond its current use to increase erythropoiesis. PMID- 11395858 TI - Tobacco risk factors: the role of the physician in protecting children from cigarettes. PMID- 11395859 TI - Use of the GFP reporter as a vital marker for Agrobacterium-mediated transformation of sugar beet (Beta vulgaris L.). AB - Molecular approaches to sugar beet improvement will benefit from an efficient transformation procedure that does not rely upon exploitation of selectable marker genes such as those which confer antibiotic or herbicide resistance upon the transgenic plants. The expression of the green fluorescent protein (GFP) signal has been investigated during a program of research that was designed to address the need to increase the speed and efficiency of selection of sugar beet transformants. It was envisaged that the GFP reporter could be used initially as a supplement to current selection regimes in order to help eliminate "escapes" and perhaps eventually as a replacement marker in order to avoid the public disquiet associated with antibiotic/herbicide-resistance genes in field-released crops. The sgfp-S65T gene has been modified to have a plant-compatible codon usage, and a serine to threonine mutation at position 65 for enhanced fluorescence under blue light. This gene, under the control of the CaMV 35S promoter, was introduced into sugar beet via Agrobacterium-mediated transformation. Early gene expression in cocultivated sugar beet cultures was signified by green fluorescence several days after cocultivation. Stably transformed calli, which showed green fluorescence at a range of densities, were obtained at frequencies of 3-11% after transferring the inoculated cultures to selection media. Cocultivated shoot explants or embryogenic calli were regularly monitored under the microscope with blue light when they were transferred to media without selective agents. Green fluorescent shoots were obtained at frequencies of 2-5%. It was concluded that the sgfp-S65T gene can be used as a vital marker for noninvasive screening of cells and shoots for transformation, and that it has potential for the development of selectable marker-free transgenic sugar beet. PMID- 11395860 TI - Purification and characterization of monoclonal antibodies against the free alpha subunit of human chorionic gonadotrophin. AB - Monoclonal antibodies (Mabs) against human chorionic gonadotropin hormone (hCG) were raised by hybridoma technology using Sp2/0 myeloma cells as fusion partner. Sixty-five percent of the total culture wells exhibited hybrid growth and 8% of the total wells (13 culture wells) contained anti-hCG secreting hybrids. A positive hybrid cell line secreting antibodies against the free alpha-subunit of hCG was cloned twice by limiting dilution method and eighty four clones were obtained that secreted monoclonal antibodies anti-alpha hCG. One of these hybridoma clones (1C4) secreting monoclonal antibodies against the free alpha subunit of hCG was selected for purification and characterization purposes. This hybridoma cell line secreted monoclonal antibodies of IgG1 subclass, which were purified by affinity chromatography on Protein A Sepharose CL-4B column with a final relative recovery of antibody activity of 75% and a purification factor of about 12. The purified preparation was analyzed by SDS-PAGE, native PAGE, and IEF. Specificity studies of this Mab revealed that it recognized specifically an epitope on the free alpha-subunits of hCG, FSH, LH, and TSH as determined by enzyme immunoassays. On the other hand, this Mab exhibited crossreactivity with other pituitary hormones either as free subunits or intact molecules as follows: alpha hCG 100%; intact hCG 1.8%; beta hCG 0.14%; alpha FSH 24.5%; intact FSH 0.8%; beta FSH 0.09%; alpha LH 20.5%; intact LH 0.9%; beta LH 0.08%; alpha TSH 50.5%; intact TSH 3.7%; beta TSH 0.07%; The affinity constant (K) of this Mab with respect to free alpha-subunit of hCG was found to be 1.5 x 10(7) I/mol as determined by the simple antibody dilution analysis method. PMID- 11395861 TI - Incorporating the ABI GeneScan analysis to a RACE-based technique for mapping multiple transcription initiation sites. AB - Determination of transcription initiation sites has commonly been performed by primer extension and RNase protection assay using radioactively labeled oligonucleotides. Recently, a protocol based on modified 5' rapid amplification of cDNA ends (RACE) with the use of fluorescently labeled primer was developed. Here, we describe the use of RACE-based technique in conjunction with the GeneScan analysis for the determination of transcription initiation sites of genes of interest. The RACE technique is based on the ligation of an adapter to both ends of the cDNAs. The gene of interest was first amplified by PCR using a gene-specific and a 5' adapter primer. Subsequently, nested PCR was performed using an internal gene-specific primer paired with a fluorescently end-labeled adapter primer. The size of the fluorescently labeled PCR products was directly determined by the ABI PRISM 377 GeneScan Analyzer. This novel approach provides an accurate, sensitive, and convenient method for mapping transcription initiation sites, especially for genes with multiple transcription initiation sites, for genes expressed at low levels, and for splice variants that display alternative splicing farther than a few hundred nucleotides downstream from the transcription initiation site. This article describes the application of this new method in the mapping of transcription initiation sites of two splice variants of rat frizzled related protein transcripts. PMID- 11395862 TI - Two-step cycle sequencing improves base ambiguities and signal dropouts in DNA sequencing reactions using energy-transfer-based fluorescent dye terminators. AB - The use of automated fluorescent DNA sequencer systems and PCR-based DNA sequencing methods plays an important role in the actual effort to improve the efficiency of large-scale DNA analysis. While dideoxy-terminators labeled with energy-transfer dyes (BigDyes) provide the most versatile method of automated DNA sequencing, premature terminations result in a substantially reduced reading length of the DNA sequence. Premature terminations are usually evidenced by base ambiguities and are often accompanied by diminished signal intensity from that point on in the sequence. I studied a two-step protocol for Taq cycle sequencing using the ABI BigDye terminator for reducing premature terminations in DNA sequences. I demonstrate that combining the annealing step with the extension step at one temperature (60 degrees C) reduces premature terminations in DNA sequences that regularly contain premature terminations when the three temperature steps are used. This modification significantly increases the number of accurately read bases in DNA sequences. PMID- 11395863 TI - Assuring the quality, safety, and efficacy of DNA vaccines. AB - Scientists in academia whose research is aimed at the development of a novel vaccine or approach to vaccination may not always be fully aware of the regulatory process by which a candidate vaccine becomes a licensed product. It is useful for such scientists to be aware of these processes as the development of a novel vaccine could be problematic owing to the starting material often being developed in a research laboratory under ill-defined conditions. This paper examines the regulatory process with respect to the development of a DNA vaccine. DNA vaccines present unusual safety considerations that must be addressed during preclinical safety studies, including adverse immunopathology, genotoxicity through integration into a vaccinees chromosomes, and the potential for the formation of anti-DNA antibodies. PMID- 11395864 TI - Production of transgenic rodents by the microinjection of cloned DNA into fertilized one-cell eggs. AB - Transgenic technologies that enable rapid movement between genotype and phenotype through specific loss-of-function, overexpression, or misexpression phenotypes will be crucial in the elucidation of gene sequences emerging from genome projects. This article describes detailed procedures for the generation of transgenic mice and rats by the injection of cloned DNA into the pronuclei of fertilized one-cell eggs. PMID- 11395865 TI - End-labeling of long DNA fragments with biotin and detection of DNA immobilized on magnetic beads. AB - To immobilize DNA fragments onto magnetic beads coated with streptavidin for isolation purpose, it is important to label one biotin molecule at one terminus of DNA fragment. After failure to label long DNA with biotin by PCR and filling in reaction, a 9.2 kb DNA was labeled with biotin by a modified ligation strategy. A simple method is also reported to detect the quantity and integrity of DNA immobilized on the magnetic beads. PMID- 11395867 TI - [Diabetology at the threshold of the 21st century]. AB - Scientific advances in the 19th and 20th centuries led to the discovery of insulin, the fundamental therapeutic means for treatment of diabetes mellitus type 1 at the onset of the twenties, to the introduction of sulfonylurea derivatives and biguanides in the fifties and sixties. The discovery of the principle of radioimmunoassay at the end of the fifties made it possible to investigate insulin secretion and to achieve a more accurate understanding of the pathogenesis of type 1 and 2 diabetes. Understanding of insulin resistance made it possible to introduce an euglycaemic hyperinsulin clamp at the end of the seventies. Insulin resistance was presented in context with metabolic syndrome X. Insulin is administered at the break of the millenium in subcutaneous injections, insulin dispensers and insulin pumps, experimentally also by the intraperitoneal and inhalatory route. In the nineties in the practice of diabetes 1 therapy ultrashort-term and finally als long-term insulin analogues were developed. For type 2 diabetes mellitus inhibitors of alpha-amylase were introduced and as a quite new group of oral antidiabetics thiazolidindiones. The possibility of 24 hour monitoring of the blood sugar level by means of a subcutaneous glucose sensor was introduced. The end of the century is characterized also by attempts to administer growth factors in the treatment of non-patent vascular obstructions in the diabetic foot syndrome. In mice and rats transformation of the ductal cell of the exocrine pancreas to the Langerhans islet cell proved successful. Further progress in diabetology will depend, similarly as in the rest of medicine, in particular on advances in cellular and molecular biology and genetics, as well as advances in microelectronics and new materials. Emphasis on the community understanding of this disease and consequential primary prevention of diabetes and secondary prevention of its complications are important. PMID- 11395866 TI - The rescue by phage display of human Fabs to gp120 HIV-1 glycoprotein using EBV transformed lymphocytes. AB - Human hybridomas secreting monoclonal antibodies in a stable manner are difficult to develop. The main difficulties are the restricted techniques for B-cell immortalization, the low number of sensitized B cells in peripheral blood, and the impossibility, for ethical reasons, to immunize humans with most antigens. Phage display has proved to be a powerful method for the generation of recombinant antibody fragments. This technology relies on the construction of recombinant Fab or scFv libraries and their display on phage M13. In order to rescue unstable B-cell clones secreting human antibodies we set up a method for the selection by phage display of human IgG fragments from Epstein-Barr virus (EBV)-transformed clones and applied it to the selection by phage display of Fabs directed against HIV-1 gp120, using a seropositive blood sample. The approach combines B-cell transformation by EBV of peripheral blood lymphocytes from a seropositive donor, preselection of specific IgG anti-gp120 producing clones, and the construction of a targeted human antibody library. In this library the percentage of heavy and light chain coding sequences expressed in Escherichia coli, amplified by a set of specific 5' primers for different antibody germ lines, was similar to that observed with the original untransformed B-cell sample. One round of panning was sufficient for the rescue of three Fabs specific for HIV-1 gp120 protein, which proves the efficiency of this technique. PMID- 11395868 TI - [Diabetes mellitus--a risk factor for cardiovascular diseases]. AB - Hyperglycemia besides typical changes of lipids, especially of LDL and HDL cholesterol and triglycerides, arterial hypertension and smoking is another factor causing greater morbidity and mortality of diabetic patients with cardiovascular disease. Significant association was found between the insulin resistance and atherosclerosis. Some studies evaluating the tight relationship between diabetes and cardiovascular diseases are selected in this overview and they demonstrate the necessity of the complex therapeutic and preventive approaches. PMID- 11395869 TI - [Current and future aspects of oral antidiabetic agents in type 2 diabetes]. AB - Type 2 diabetes mellitus is characterized by insulin deficiency but in particular by insulin resistance. Patients where it is not possible to achieve positive results within 4-12 weeks by optimalization of the lifestyle are candidates for treatment with oral antidiabetics. At present the following main groups of oral antidiabetics are discussed: insulin secretagogues (SU derivatives and methiglinide derivatives), biguanides (Metformin), alpha-glucosidase inhibitors (acarbose, miglitol) and insulin sensitizers (thiazolindiones). Traditional SU therapy improves the insulin plasma levels by releasing insulin from the pancreas. This implies further stress on the b-cells and the function of these cells declines reversibly. Biguanides, such as metformin, are effective substances reducing the blood sugar level, they are however associated with the problem of tolerability and are contraindicated in some diabetics. A new approach to the treatment of type 2 diabetes are thiasolinediones, insulin-sensitizing substances, the molecular basis of their action being via activation of PPAR gamma-nuclear receptors with subsequent change in expression of genes participating in carbohydrate and lipid metabolism. PMID- 11395871 TI - [Education of diabetics--an essential part of therapy]. PMID- 11395870 TI - [Role of insulin therapy in the treatment of type 2 diabetes]. PMID- 11395872 TI - [The phenomenon of unawareness of hypoglycemia]. AB - The incidence of hypoglycaemic episodes in patients with type 1 diabetes mellitus (DM1) is frequent. They experience 1-2 symptomatic hypoglycaemias per week. 10 20% of the patients suffer from at least one severe hypoglycaemia per year. The incidence of severe hypoglycaemia in a group of intensely treated patients is about three times as high as compared with standard treatment and 55% of all episodes occur during sleep. Prevention of hypoglycaemia, and restoration of euglycaemia resp. include disappearance of metabolic effects of insulin and activation of glucose contraregulating systems among which there is a certain hierarchy. The phenomenon of unaware hypoglycaemia (FNH) is defined as failure to diagnose autonomous warning symptoms, and their non-appearance before the development of neuroglycopenia. The incidence of FNH in the population with DM1 is frequent. Based on a standardized insulin diffusion test 26% of patients with DM1 suffer from it which means that every fourth patient is affected. In the pathogenesis of FNH various factors and mechanisms were suggested as predisposing. Most probably a defect at the level of the central nervous system is involved caused by: 1. a reduced ability to recognize the decline of the blood sugar level by the CNS (altered function of the hypothalamic glucostat and/or glucose transport across the haematoencephalic barrier), 2. reduced secretion of neurotransmitters, 3. reduced tissue response to adequate neurotransmitter secretion. The theory of etiopathogenesis must explain and define its association with the persistence of diabetes mellitus, strict metabolic control, autonomous neuropathy and repeated episodes of hypopglycaemia. The contemporary hypothesis of the pathogenesis of FNH is the mechanism of repeated frequent hypoglycaemia which leads to general adaptative changes at the level of the CNS by increased glucose transport across the haematoencephalic barrier which leads to reduced hormonal responses and reduction of symptoms. Thus a dangerous circulus vitiosus is created where hypoglycaemia induces unawareness of hypoglycaemia. This condition is at least partly reversible. The presence of FNH should influence the decision of the physician before using an intensified insulin regimen in diabetics. PMID- 11395873 TI - [Care of diabetics in an acute state]. PMID- 11395874 TI - [Microalbuminuria--a risk factor for diabetic nephropathy and cardiovascular disease]. AB - In developed countries diabetics patients are the most numerous group with renal replacement therapy (USA 34%). The main and diagnostically irreplaceable criterion of incipient diabetic nephropathy is microalbuminuria which is usually associated with hypertension and poor glycaemic compensation. With advancing microalbuminuria progresses diabetic retinopathy and neuropathy. The increased transcapillary albumin escape rate and changes of some haemocoagulation factors in diabetics patients with microalbuminuria indicate that endothelial dysfunction is involved. In type 1 diabetes microalbuminuria is an indicator of increased mortality in which participate in particular cardiovascular diseases and to a minor extent renal failure. In type 2 diabetes microalbuminuria is an independent risk of generalized vascular disease. Microalbuminuria is also in non-diabetic subjects with hypertension associated with abnormalities such as impaired glucose tolerance and insulin resistance, an unflavourable lipidogram and altered diurnal blood pressure rhythm. The results of a coronarographic investigation revealed that the risk of severe coronary artery disease is more than double in subjects with microalbuminuria. Hypertension and hypercholesterolaemia are causal risk factors of cardiovascular diseases and concurrent microalbuminuria implies a higher expression of already existing microvascular damage in hormonal and metabolic disorders with an atherogenic potential. PMID- 11395875 TI - [The diabetic foot syndrome--one of the most serious complications in diabetics]. AB - The diabetic foot syndrome is defined according to WHO as ulceration of the foot (distally from the ankle and including the ankle) associated with neuropathy and different grades of ischaemia and infection. It is the most frequent cause of hospitalization of diabetic patients. It is one of the economically most demanding complications of diabetes. According to international consensus on the diabetic foot syndrome should be organized at the level of general practitioners as well as at the level of special diabetological centres. The basic organizational unit of multidisciplinary team care of patients with the diabetic foot syndrome is foot clinic. The diabetologist is the main coordinator of the foot clinic and responsible for all stages of treatment of patients with the diabetic foot syndrome. Essential members of the team taking care of patients with the diabetic foot syndrome are podiatric nurses, a specialized general and vascular surgeon, an intervention radiologist and prosthetist. In the diagnosis of the diabetic foot syndrome vascular examination is most important, neurological preliminary examination, assessment of the compensation of diabetes and the presence of infection. In treatment non-weight bearing, better vascularization, treatment of infection, improvement of the metabolic condition and prevention of reulcerations are most important. PMID- 11395876 TI - [Stress-induced hypertension and diabetes mellitus]. AB - Hypertension and diabetes are the basic risk factors of atherosclerosis and its complications. At present new associations are sought which will enable us to describe more satisfactorily the mutual relationship of hypertension, metabolic disorders and cardiovascular disease. One of the systems involved in all substantial physiological processes is the autonomic nervous system. Stimulation of the sympathetic nervous system by chronic stress causes in addition to an elevated pulse rate and cardiac minute output also activation of another important pressor mechanism--the renin-angiotensin-aldosterone system. Increased activity of the sympathetic nervous system plays a part also in the development of impaired glucose and lipid metabolism, which are very frequent in hypertonic subjects. Hyperinsulinaemia, hypertriglyceridaemia and reduced HDL-cholesterol concentration are associated with a decline of the insulin capacity to take up glucose and deposit glycogen and together with a raised blood pressure create the so-called metabolic syndrome of insulin resistance (syndrome X, Reaven's syndrome). PMID- 11395877 TI - [Special aspects of diagnosis and therapy of diabetes in liver diseases]. AB - The core of this review are questions dealing with links between diabetes mellitus and liver diseases. Based on the literature and on their own observations the authors conclude that a liver disease may induce and/or worsen the development of type-2 diabetes mellitus (so called hepatogenic diabetes) and vice versa, a diabetes may result in a liver damage, e.g. simple steatosis or even non-alcoholic-steatohepatitis. The diagnosis of diabetes in hepatopathy is defined according to the same criteria as in patients without any hepatic involvement. The treatment is based on adequate food intake without alcohol and on restricted physical activity. Complementary insulin treatment is recommended whenever the BG-concentrations in a daily profile exceed 10.0 mmol/l. Oral antidiabetic drugs should be used only if they result in a marked improvement of diabetes control and no signs of liver damage appear. PMID- 11395878 TI - [Are ultrasonic images in diabetics different?]. AB - The authors evaluated the results assembled in 5397 patients where between Jan. 1 1999 and Oct. 31 2000 sonographic examinations of the abdominal cavity and retroperitoneum were made with the objective to assess whether there are any statistically significant differences of results in diabetic patients, as compared with a group without this disease. The group of patients was divided into a sub-group of 4287 patients without diabetes and a sub-group of 1100 diabetics. For statistical evaluation of the significance of differences in the incidence of the investigated parameters Fisher's exact test was used. The image of "light liver" was significantly more frequent in diabetics type 1 and 2, as compared with non-diabetics (p < 0.001). The sonographic picture, consistent with the diagnosis of cirrhosis of the liver, was at the same level of significance more frequent in non-diabetics, similarly as the incidence of haemangioma. The finding of cholecystolithiasis and the number of patients with a history of CHCE on account of cholecysolithiasis was significantly higher (p < 0.05) only in type 1 diabetics as compared with non-diabetics. The incidence of sonographic changes consistent with acalculous cholecystitis was statistically higher in both groups of diabetics (p < 0.001), as compared with non-diabetics. On examination of the pancreas only the incidence of changes consistent with acute or chronic pancreatitis was significantly higher (p < 0.05) in the group of type 2 diabetics as compared with non-diabetics. Evaluation of sonographic findings of the kidneys revealed statistically significant differences only in the higher incidence of cysts in the group of type 2 diabetics as compared with type 1 diabetics and as compared with non-diabetics (p < 0.01). The impact of the presented findings and their comparison with data reported in the literature is discussed. PMID- 11395879 TI - [Transcutaneous oximetry in the diagnosis of ischemic disease of the lower extremities in diabetics]. AB - In diabetic patients with ischaemia of the lower extremities and mediocalcinosis the diagnosis of ischaemia is still difficult. One of the methods which give an objective idea of the microcirculation is transcutaneous oximetry. In the submitted paper the authors explain the principle and problems of assessment with recommendations for the practical application of the method, in particular in indication for angiography of the lower extremities in the presence of the diabetic foot syndrome with regard to probable spontaneous healing of defects and application of the method for following up the effect of revascularization operations. PMID- 11395880 TI - [Preoperative and postoperative care in diabetics]. AB - The submitted paper should serve as a basis for recommended procedures of preoperative and postooperative care of diabetics. It contains the algorithm of preoperative preparation of diabetics focused on quantification of the risk of surgery. The diabetologist is the consultant of the surgeon who participates in the preparation of the patient for urgent as well as elective surgery. The procedures differ with regard to the type of diabetes. The authors mention in more detail the problem of preoperative care of operated diabetics and finally also monitoring and therapy of diabetics during the postoperative period. PMID- 11395881 TI - Incidence of acute renal failure in the intensive care unit. PMID- 11395882 TI - Transesophageal echocardiography in the ICU. PMID- 11395883 TI - Antibiotic therapy in acute renal failure. PMID- 11395884 TI - The renal effects of noradrenaline and dopamine. PMID- 11395885 TI - The role of diuretic agents in the management of acute renal failure. PMID- 11395886 TI - Renal replacement therapy (RRT) in the ICU: criteria for initiating RRT. PMID- 11395887 TI - Severity scores and outcomes with acute renal failure in the ICU setting. PMID- 11395888 TI - Answers from the first international course on critical care nephrology questionnaire. PMID- 11395889 TI - Peritoneal dialysis in acute renal failure of adults: the safe, effective, and low-cost modality. PMID- 11395890 TI - Risk factors for acute renal failure in the intensive care unit. PMID- 11395891 TI - Selected practical aspects of intermittent hemodialysis in acute renal failure patients. PMID- 11395892 TI - Continuous renal replacement techniques. PMID- 11395893 TI - Hybrid renal replacement modalities for the critically ill. PMID- 11395894 TI - Acute dialysis quality initiative (ADQI). PMID- 11395895 TI - The kidney in shock. PMID- 11395896 TI - Vascular access for extracorporeal renal replacement therapies in the intensive care unit in clinical practice. PMID- 11395897 TI - Anticoagulation in continuous renal replacement therapy. PMID- 11395898 TI - Factors influencing therapy delivery in acute dialysis. PMID- 11395899 TI - Dialysate and substitution fluids for patients treated by continuous forms of renal replacement therapy. AB - The development of CRRT from simple arteriovenous ultrafiltration with a treatment using 50 liters a day, has necessitated the development of specialized dialysate/replacement solutions. As the volumes used are large, quality control is vitally important not only in terms of electrolyte composition, but also preventing bacterial and endotoxin contamination. Lactate remains the standard anionic base, as this increases fluid storage time and reduces the risk of bacterial contamination. Some centers use citrate anticoagulation, and the citrate load provides an adequate supply of anionic base to control metabolic acidosis. However, patients receiving very high volume exchanges, or those with severe tissue acidosis and/or liver failure, may be unable to convert the lactate, or citrate load effectively, resulting in hyperlactatemia or hypercitratemia and acidosis. In these circumstances, bicarbonate-based fluids are advantageous. The use of bicarbonate-based dialysate/replacement fluid during CRRT does not mask lactate overproduction, and lactate remains a reliable marker of tissue oxygenation in patients treated by CRRT. Bicarbonate CRRT alone does not treat metabolic acidosis, but allows better control of acidosis, thereby allowing time for the institution of other therapies designed to reverse the underlying cause. PMID- 11395900 TI - Machines for continuous renal replacement therapy. PMID- 11395901 TI - Fluid management in CRRT. AB - Fluid management with CRRT requires an understanding of the principles of fluid removal and fluid balance. Although these appear to be similar to intermittent hemodialysis, there are significant differences. In order to utilize these techniques to their full ability, a variety of strategies can be used. No matter which method is used it is imperative that the goals for fluid management be well defined and monitoring for errors be a part of the protocol. PMID- 11395902 TI - Update on drug sieving coefficients and dosing adjustments during continuous renal replacement therapies. PMID- 11395903 TI - Sepsis: a pro- and anti-inflammatory disequilibrium syndrome. AB - Severe sepsis and probably most prolonged critical illnesses reflect a paradox of combined increased activation and depression of the immune apparatus. The increased activation of the inflammatory response is evidenced from the increased levels of circulating proinflammatory cytokines in the blood, increased endothelial activation with increased expression of inducible nitric oxide synthase, and increased de novo CD11b expression on circulating immune effector cells, such as PMNs, monocytes and lymphocytes. However, coexisting with this proinflammatory process is a profound anti-inflammatory state characterized by increased circulating levels of anti-inflammatory species that both directly block the binding of proinflammatory stimuli to their cell surface receptors (IL 1ra, soluble TNF receptors) and also induce an anti-inflammatory state on their own (IL-10, TFG-beta). This humoral anti-inflammatory state is mirrored at the cellular levels by decreased monocyte ability to process antigen, characterized by a reduced HLA-DR expression and impaired PMN upregulation in response to clearly proinflammatory stimuli. Accordingly, severe sepsis reflects a combined pro- and anti-inflammatory state. Both the pro- and anti-inflammatory arms have protective and destructive aspects, making their modulation by treatment less predictable than if their actions were purely beneficial or detrimental. PMID- 11395904 TI - Extracorporeal blood purification therapy for sepsis and systemic inflammation: its biological rationale. AB - EBPTs represent a promising new approach to the adjuvant treatment of severe sepsis, septic shock and MODS. Their technology is rapidly evolving and pilot animal and human studies are now taking place to prepare the territory for the first large randomized controlled trial. The rationale for EBPT is reasonable and the initial data are encouraging. The correct technology and molecular targeting, however, are still being explored. Once the best technology has been determined, it is likely that phase II and phase III trials will be performed to test the hypothesis that these therapies can indeed alter mortality in severe inflammatory multiorgan dysfunction. PMID- 11395905 TI - High-volume hemofiltration. PMID- 11395907 TI - Plasmapheresis in sepsis. PMID- 11395906 TI - Coupled plasma filtration adsorption in the treatment of septic shock. PMID- 11395908 TI - Should we target signal pathways instead of single mediators in the treatment of sepsis? PMID- 11395909 TI - Molecular mechanisms underlying combined kidney-lung dysfunction during acute renal failure. PMID- 11395910 TI - Endotoxin removal by toraymyxin. PMID- 11395911 TI - Thrombotic microangiopathies in the intensive care unit: many questions, some answers. PMID- 11395912 TI - Role of the renal biopsy in acute renal failure. PMID- 11395913 TI - Pathophysiology of ischemic acute renal failure. PMID- 11395914 TI - Imaging techniques in acute renal failure. PMID- 11395915 TI - Hemodynamic monitoring in the ICU. PMID- 11395916 TI - Lymphatic filariasis. PMID- 11395917 TI - Malaria information service, Kenya. PMID- 11395918 TI - Leprosy. Global target attained. PMID- 11395919 TI - Omega-3 polyunsaturated fatty acids and skeletal health. AB - This minireview on skeletal biology describes the actions of prostaglandins and cytokines involved in the local regulation of bone metabolism, it documents the role of lipids in bone biology, and it presents relationships between fatty acids and other factors that impact skeletal metabolism. The data presented herein show consistent and reproducible beneficial effects of omega-3 (n-3) fatty acids on bone metabolism and bone/joint diseases. Polyunsaturated fatty acids modulate eicosanoid biosynthesis in numerous tissues and cell types, alter signal transduction, and influence gene expression. These effects have not been explored in the skeletal system. Future research on n-3 fatty acids in bone biology should focus on the following two aspects. First, the further elucidation of how n-3 fatty acids alter biochemical and molecular processes involved in bone modeling and bone cell differentiation, and second, the evaluation of the potential pharmaceutical applications of these nutraceutical fatty acids in maintaining bone mineral status and controlling inflammatory bone/joint diseases. PMID- 11395920 TI - The role of n-3 fatty acids in gestation and parturition. AB - Preterm birth is the most common cause of low infant birth weight and infant morbidity and mortality. Evidence from human and animal studies indicates that essential fatty acids of both the n-3 and n-6 series, and their eicosanoid metabolites, play important and modifiable roles in gestational duration and parturition, and n-3 fatty acid intake during pregnancy may be inadequate. Prostaglandins (PG) of the 2-series are involved in parturition and connective tissue remodeling associated with cervical maturation and rupture of membranes. In the absence of infections, preterm birth is characterized by lower reproductive tissue PG production and decreased inducible cyclooxygenase expression. Women who deliver prematurely have increased pools of n-6 fatty acid and decreased n-3 fatty acids, despite the lower PG production. Several human pregnancy supplementation trials with n-3 fatty acids have shown a significant reduction in the incidence of premature deliver and increased birth weight associated with increased gestational duration. Supplementation with long chain n 3 fatty acids such as docosahexaenoic acid may be useful in prolonging the duration of gestation in some high-risk pregnancies. Evidence presented in this review is discussed in terms of the roles of dietary n-3 and n-6 fatty acids in gestation and parturition, mechanisms by which they may influence gestational duration and the human trials suggesting that increased dietary long-chain n-3 fatty acids decrease the incidence of premature delivery. PMID- 11395921 TI - Mesenchymal stem cells. AB - Within the bone marrow stroma there exists a subset of nonhematopoietic cells referred to as mesenchymal stem or mesenchymal progenitor cells. These cells can be ex vivo expanded and induced, either in vitro or in vivo, to terminally differentiate into osteoblasts, chondrocytes, adipocytes, tenocytes, myotubes, neural cells, and hematopoietic-supporting stroma. The multipotential of these cells, their easy isolation and culture, as well as their high ex vivo expansive potential make these cells an attractive therapeutic tool. In this work we will review the information dealing with the biology of mesenchymal progenitors as it has been revealed mainly by ex vivo studies performed with bone marrow-derived cells. The discussed topics include, among others, characteristics of mesenchymal progenitors, evidence for the existence of a vast repertoire of uncommitted and committed progenitors both in the bone marrow and in mesenchymal tissues, a diagram for their proliferative hierarchy, and comments on mobilization, microenvironment, and clinical use of mesenchymal progenitors. Despite the enormous data available at molecular and cellular levels, it is evident that a number of fundamental questions still need to be resolved before mesenchymal progenitors can be used for safe and effective clinical applications in the context of both cell and gene therapies. PMID- 11395922 TI - Selected features of nonendocrine pancreatic cancer. AB - We survey some interesting features of gene expression in nonendocrine pancreatic cancer, the response to some less widely known agents as they impact on pancreatic cell proliferation and programmed death, and several developing approaches to therapy. The proliferative and cellular suicide responses of Panc-1 cells to the free radical spin trap, NTBN, and to the 5-lipoxygenease inhibitor, MK 886, the latter assessed with CLONTECH Atlas Human cDNA Array 1, are reviewed. Difficulties in identifying those factors whose suppression or augmentation could result in inhibition of malignantly transformed cell properties are considered. PMID- 11395923 TI - Acute, nongenomic vasodilatory action of estradiol is attenuated by chronic estradiol treatment. AB - Deficiency of estradiol or chronic estrogen treatment may alter the responses to this hormone in many tissues. A possible interaction between the acute nongenomic and the chronic effects of estradiol on microvessels have not been investigated yet. In the present study we have investigated whether acute in vitro vasodilatory action of estradiol on a small artery is altered by chronic estradiol pretreatment. Female rats were surgically ovariectomized and subjected to either estradiol replacement therapy (estradiol propionate, 450 micrograms/kg/week) or vehicle administration for 5 weeks. Cylindrical segments of the saphenous artery were studied using videocomputerized microarteriography in vitro. Estradiol, in concentrations of 10(-6) to 10(-4) M relaxed norepinephrine precontracted vessel segments in a dose-dependent manner. Magnitude of relaxation observed in arteries of estradiol replaced animals was significantly smaller at all concentrations than that of nonreplaced ovariectomized rats; maximal relaxation in the control ovariectomized group was 64.5% +/- 3.6%, while it was 34.3% +/- 4.2% only in the ovariectomized and estradiol replaced group (P < 0.001). Comparison of acute relaxations in response to papaverine and nifedipine failed to prove a reduced activity of the general relaxation machinery in estradiol replaced animals. We conclude that chronic estradiol replacement can downregulate the acute nongenomic vasorelaxation effect of this hormone in small arteries of ovariectomized rats. PMID- 11395924 TI - Na+ effects on mitochondrial respiration and oxidative phosphorylation in diabetic hearts. AB - Intracellular Na+ is approximately two times higher in diabetic cardiomyocytes than in control. We hypothesized that the increase in Na+i activates the mitochondrial membrane Na+/Ca2+ exchanger, which leads to loss of intramitochondrial Ca2+, with a subsequent alteration (generally depression) in bioenergetic function. To further evaluate this hypothesis, mitochondria were isolated from hearts of control and streptozotocin-induced (4 weeks) diabetic rats. Respiratory function and ATP synthesis were studied using routine polarography and 31P-NMR methods, respectively. While addition of Na+ (1-10 mM) decreased State 3 respiration and rate of oxidative phosphorylation in both diabetic and control mitochondria, the decreases were significantly greater for diabetic than for control. The Na+ effect was reversed by providing different levels of extramitochondrial Ca2+ (larger Ca2+ levels were needed to reverse the Na+ depressant effect in diabetes mellitus than in control) and by inhibiting the Na+/Ca2+ exchanger function with diltiazem (a specific blocker of Na+/Ca2+ exchange that prevents Ca2+ from leaving the mitochondrial matrix). On the other hand, the Na+ depressant effect was enhanced by Ruthenium Red (RR, a blocker of mitochondrial Ca2+ uptake, which decreases intramitochondrial Ca2+). The RR effect on Na+ depression of mitochondrial bioenergetic function was larger in diabetic than control. These findings suggest that intramitochondrial Ca2+ levels could be lower in diabetic than control and that the Na+ depressant effect has some relation to lowered intramitochondrial Ca2+. Conjoint experiments with 31P NMR in isolated superfused mitochondria embedded in agarose beads showed that Na+ (3-30 mM) led to significantly decreased ATP levels in diabetic rats, but produced smaller changes in control. These data support our hypothesis that in diabetic cardiomyocytes, increased Na+ leads to abnormalities of oxidative processes and subsequent decrease in ATP levels, and that these changes are related to Na+ induced depletion of intramitochondrial Ca2+. PMID- 11395925 TI - Reduced levels of thyroid hormones, insulin, and glucose, and lower body core temperature in the growth hormone receptor/binding protein knockout mouse. AB - The mechanisms that are responsible for the extension of lifespan in the mouse with targeted disruption (knockout [KO]) of the growth hormone (GH) receptor/binding protein (GHR-KO) are unknown. However, in the long-living Ames dwarf mouse, blood glucose and body core temperature (Tco) are consistently lower than in normal mice. In addition, insulin levels are reduced and corticosterone levels are elevated in male dwarfs. These functional alterations, similar to those seen in animals under caloric restriction, have not been proven to be causally related to the extension of lifespan, but they do provide some insight into what traits may be necessary for long life. Therefore, to investigate which of these parameters are similarly affected in two genetically unrelated, yet similarly long-living mouse models, we measured Tco, thyroid hormones (triiodothyronine [T3] and thyroxine [T4]), and insulin, in addition to morning and afternoon levels of glucose and corticosterone, in young adult male and/or female GHR-KO mice and their normal siblings. Tco in GHR-KO mice was numerically reduced throughout the 24-hr period; however, these differences were only significant 4 hr prior to lights-off (14:00 hr), immediately after lights-off (18:00 hr), and during the 3 hr preceding lights on (03:00 to 06:00 hr). GHR-KO mice had significantly reduced levels of T3 and T4, while the ratio of these hormones was similar to that in normal mice. Insulin levels in GHR-KO mice were lower than in normal mice; levels in male GHR-KO mice were below the detectable limits of the assay used. Glucose levels in GHR-KO mice (male and females) were lower than in normal mice in measurements taken in both morning and afternoon; however, these differences arose from consistent reductions in males, as morning glucose levels in GHR-KO females were similar to those of normal mice. Corticosterone levels measured in blood plasma collected under basal (nonstressed) conditions showed sex-related alterations. Basal corticosterone levels in female GHR-KO mice were similar to normal females, while those in male GHR-KO mice were higher than in normal males in the afternoon. Corticosterone levels in stressed GHR-KO females were similar to those measured in stressed normal females. These data show that the long-living GHR-KO mouse shares a reduction in glucose, insulin, thyroid hormones, and Tco with the Ames dwarf mouse. Reductions in these parameters may be important to the underlying mechanisms of delayed aging in these animals. PMID- 11395926 TI - Liposomes as possible carriers for lactoferrin in the local treatment of inflammatory diseases. AB - Liposomes prepared from naturally occurring biodegradable and nontoxic lipids are good candidates for local delivery of therapeutic agents. Treatment of arthritis by intra-articular administration of anti-inflammatory drugs encapsulated in liposomes prolongs the residence time of the drug in the joint. We have previously shown that intra-articular injection of human lactoferrin (hLf), a glycoprotein that possesses anti-inflammatory and antimicrobial activities, into mice with collagen-induced arthritis reduces inflammation. We have now investigated the possibility of using liposome-entrapped hLf as a delivery system to prolong hLf retention at sites of local inflammation such as the rheumatoid joint. Entrapment of hLf in negatively charged liposomes enhanced its accumulation in cultured human synovial fibroblasts from rheumatoid arthritis (RA) patients, compared with positively charged formulations or free protein. However, in the presence of synovial fluid, positively charged liposomes with entrapped hLf were more stable than the negatively charged formulations. In vivo experiments in mice with collagen-induced arthritis showed that the positive liposomes were more efficient in prolonging the residence time of hLf in the inflamed joint as compared with other liposomes. Thus, the amount of hLf retained in the joint after 2 hr was 60% of the injected dose in the case of positive liposomes and only 16% for negative pH-sensitive liposomes. The results suggest that entrapment of hLf in positively charged liposomes may modify its pharmacodynamic profile and be of therapeutic benefit in the treatment of RA and other local inflammatory conditions. PMID- 11395927 TI - Agouti signaling protein stimulates islet amyloid polypeptide (amylin) secretion in pancreatic beta-cells. AB - Ectopic overexpression of the murine agouti gene results in yellow coat color, obesity, hyperinsulinemia, and type II diabetes. We have shown the human homologue of agouti (agouti signaling protein; ASP) to regulate human adipocyte metabolism and lipid storage via a Ca(2+)-dependent mechanism. We have also demonstrated agouti expression in human pancreas, and that ASP stimulates insulin release via a similar Ca(2+)-dependent mechanism. Plasma amylin is also elevated in agouti mutant mice. Amylin is cosecreted with insulin from beta-cells, and overexpression of human amylin in beta-cells in yellow agouti mutant mice resulted in accelerated pancreatic amyloid deposition, severely impaired beta cell function, and a diabetic phenotype. We report here that ASP stimulates amylin release in both the HIT-T15 beta-cell line and human pancreatic islets in the presence of a wide range of glucose concentrations (0-16.7 mmol/L), similar to its effect on insulin release; this effect was blocked by 30 mumol/L nitrendipine, confirming a Ca(2+)-dependent mechanism. Accordingly, ASP stimulation of amylin release may serve as a compensatory system to regulate blood glucose in yellow agouti mutants. PMID- 11395928 TI - Differing effects of endocrine-disrupting chemicals on basal and FSH-stimulated progesterone production in rat granulosa-luteal cells. AB - Previous studies have shown that the phytoestrogen, genistein, inhibits basal and forskolin-stimulated progesterone synthesis in rat granulosa-luteal cells. Genistein, however, not only binds and activates the estrogen receptor (ER), but is also a potent inhibitor of tyrosine kinase. In these studies we have compared the effects of estradiol, two other phytoestrogens, apigenin and coumarin, the pesticide, [2-(chlorphenyl)-2-(4-chlorphenyl)-1,1,1-trichlorethan] (2,4'DDT), and the industrial chemical, 4-octyl-phenol, on basal and follicle stimulating hormone (FSH)-stimulated progesterone production in the same experimental system. Only a supraphysiological dose of estradiol (10(-5) M) significantly inhibited basal and forskolin-stimulated progesterone production in granulosa-luteal cells, but had no effect on FSH-stimulated production. In contrast, apigenin, DDT, and octyl-phenol stimulated basal progesterone production at doses around 10(-8) to 10(-7) M, but this effect was reversed at higher doses. Coumarin was without effect. Like basal production, the two phytoestrogens had opposing effects on FSH stimulated progesterone production. Genistein at 10(-5) M was inhibitory, while apigenin significantly potentiated the response at 19(-7) M. In contrast, DDT had no effect on the FSH-induced response, though 10(-7) M octyl-phenol nearly doubled the response. While all these chemicals are known to interact with the estrogen receptor to a greater or lesser extent, these studies suggest that like genistein, these different endocrine-disrupting chemicals may have other actions apart from those on the estrogen receptor. PMID- 11395929 TI - Black tea extract, thearubigin fraction, counteracts the effect of tetanus toxin in mice. AB - The aim of this study was to find an inactivating substance for tetanus toxin in natural foodstuff. Tetanus toxin (4 micrograms/ml) abolished indirect twitches in In vitro mouse phrenic nerve-diaphragm preparations within 2.5 hr. Hot water infusion of black tea mixed with tetanus toxin blocked the inhibitory effect of the toxin. Mixing the toxin with thearubigin fraction extracted from black tea infusion produced an identical result. Furthermore, thearubigin fraction mixed with the toxin protected against the in vivo paralytic effect of the toxin. Thearubigin fraction had no protective effect on other toxins, such as tetrodotoxin and saxitoxin. The specific binding of [125I]tetanus toxin to rat cerebrocortical synaptosomes was inhibited by mixing iodinated toxin with thearubigin fraction. These results imply that thearubigin fraction counteracts the effect of tetanus toxin by binding with toxin, and also suggest that this fraction may be able to apply for prophylaxis of tetanus. PMID- 11395930 TI - Partial restoration of lutropin activity by an intersubunit disulfide bond: implications for structure/function studies. AB - Gonadal function is controlled by lutropins and follitropins, heterodimeric cystine knot proteins that have nearly identical alpha-subunits. These heterodimeric proteins are stabilized by a portion of the hormone-specific beta subunit termed the "seatbelt" that is wrapped around alpha-subunit loop 2 (alpha 2). Here we show that replacing human chorionic gonadotropin (hCG) alpha 2 residue Lys51 with cysteine or alanine nearly abolished its lutropin activity, an observation that implies that alpha Lys51 has a key role in hormone activity. The activity of the heterodimer containing alpha K51C, but not that containing alpha K51A, was increased substantially when beta-subunit seatbelt residue beta Asp99 was converted to cysteine. As had been reported by others, heterodimers containing alpha K51C and beta D99C were crosslinked by a disulfide. The finding that an intersubunit disulfide restored some of the activity lost by replacing alpha Lys51 suggests that this residue is not crucial for receptor binding or signaling and also that hCG and related hormones may be particularly sensitive to mutations that alter interactions between their subunits. We propose the unique structures of hCG and related family members may permit some subunit movement in the heterodimer, making it difficult to deduce key residues involved in receptor contacts simply by correlating the activities of hormone analogs with their amino acid sequences. PMID- 11395931 TI - Leptin stimulates gonadotropin releasing hormone release from cultured intact hemihypothalami and enzymatically dispersed neurons. AB - Leptin is a peptide released by adipocytes that has profound effects on central regulation of body metabolism. The present study represents an investigation into leptin effects on hypothalamic control of reproductive function, specifically on GnRH release. Adult male rats (gonadectomized or sham-operated) were used as donors of hypothalamic tissue that was used as intact hemihypothalami or as enzymatically dispersed hemihypothalami in a perifusion culture system. Continuous samples were collected at 10-min intervals for 8 to 10 hr and were assayed to measure temporal changes in GnRH release in response to various doses of leptin infused into the perifusion chambers. Leptin at the highest dose (10( 8) M) resulted in consistent and significant stimulation of GnRH release. There were no effects of treatment for surgical preparation (gonadectomy versus sham) or tissue preparation (intact versus dispersed hemihypothalami). The results of this study support the hypothesis that leptin plays a direct stimulatory role in the regulation of GnRH release. This study describes an important step in our understanding of the mechanism that connects changes in basal metabolism with reproductive function. These results indicate an intact interneuronal network is unnecessary for these leptin effects, but does not exclude a role for interneuronal networks in this regulatory pathway. PMID- 11395932 TI - Anisodamine inhibits shiga toxin type 2-mediated tumor necrosis factor-alpha production in vitro and in vivo. AB - Cytokines, in particular tumor necrosis factor (TNF), appear to be necessary to develop the pathological process of Shiga toxin-producing Escherichia coli (STEC) infection. In this study we examined the effect of anisodamine, a vasoactive drug, on TNF-alpha production in Shiga toxin type 2 (Stx2)-stimulated human monocytic cells in vitro and in Stx2-injected mice sera in vivo. Human monocytes and THP-1 cells were stimulated by Stx2 (1-100 ng/ml) with or without anisodamine addition (1-400 micrograms/ml). For in vivo evaluations, C57BL/6 mice were given a single intraperitoneal injection of anisodamine (6-50 mg/kg) or saline after intraperitoneal injection of Stx2 (50 ng/kg). The results showed that anisodamine suppressed Stx2-induced TNF-alpha production in a dose- and time-dependent manner. Anisodamine also suppressed Stx2-induced TNF-alpha mRNA expression. Further study showed that endogenous prostaglandin E2 may be involved in this inhibitory effect. In contrast to TNF-alpha mRNA, anisodamine at concentrations as high as 400 micrograms/ml did not decrease Stx2-induced IL-1 beta and IL-8 mRNA levels. In addition, anisodamine (> 50 micrograms/ml) increased Stx2 stimulated THP-1 cell viability. Levels of TNF-alpha in anisodamine-treated mice sera were significantly lower than those in the saline-treated group 1.5 and 24 hr after Stx2 injection. Anisodamine induced a lower percentage of death in Stx2 injected mice. Taken together, our results indicate that anisodamine has an important regulatory effect on Stx2-induced TNF-alpha production in vitro and in vivo. The present study suggested that this drug should be further investigated for its effects on Stx2-mediated diseases in humans. PMID- 11395933 TI - Cadmium effects on hypothalamic-pituitary-testicular axis in male rats. AB - This study analyzes cadmium effects at the hypothalamic-pituitary-testicular axis. Male rats were given cadmium during puberty or adulthood. Cadmium exposure through puberty increased norepinephrine content in all hypothalamic areas studied, but not in the median eminence. Metal exposure increased serotonin turnover in median eminence and the anterior hypothalamus, while decreased it in mediobasal hypothalamus. Also, decreased plasma levels of testosterone were found. Cadmium exposure during adulthood increased norepinephrine content in posterior hypothalamus and decreased the neuro-transmitter content in anterior and mediobasal hypothalamus. Decreased circulating levels of luteinizing hormone (LH) and testosterone and increased plasma follicle stimulating hormone (FSH) levels were also observed. Cadmium accumulated in all analyzed tissues. Various parameters showed age-dependent changes. These data suggest that cadmium globally effects hypothalamic-pituitary-testicular axis function by acting at the three levels analyzed and that an interaction between cadmium exposure and age emerge. PMID- 11395934 TI - Changes in substance P content at the hypothalamic-pituitary axis during the Wallerian degeneration of peripheral sympathetic neurons after superior cervical ganglionectomy in male rats: effect of hyperprolactinemia. AB - The effects of Wallerian degeneration of the peripheral sympathetic neurons projecting to the hypothalamus on the mechanism of interaction between prolactin and substance P (SP) were examined. The effects of superior cervical ganglionectomy (SCGx) on SP content in various hypothalamic regions and in the hypophysis were evaluated in control and hyperprolactinemic rats. Male rats that received pituitary transplants at the age of 5 days and age-matched sham-operated controls were used. Pituitary grafting significantly increased circulating values of prolactin, as did SCGx. In hyperprolactinemic rats, SCGx partially decreased plasma prolactin levels. Neonatal hyperprolactinemia decreased SP content in the anterior (AH) and posterior (PH) hypothalamus and in the median eminence (ME), but increased it in the mediobasal hypothalamus (MBH). Acute SCGx significantly increased SP in the MBH, PH, and ME. SCGx in hyperprolactinemic animals further increased SP content in MBH. In the ME and Ah, SCGx in pituitary grafted rats decreased SP content as compared with the controls. In the pituitary gland (PG), SCGx only decreased SP content in hyperprolactinemic, but not in control rats. An interaction between peripheral nor-adrenergic neurons and prolactin to regulate SP within the hypothalamus was positive in the MBH, AH, ME, and PG, but not in the PH. These data indicate the existence of interactive mechanisms between prolactin and the peripheral sympathetic neurons to regulate SP content at the hypothalamic-pituitary axis. Interrelationships between prolactin and SP were also observed. PMID- 11395935 TI - MMR vaccination: how reassuring is reassurance? PMID- 11395936 TI - Henry Atkins M.D. of Great Berkhampstead for thirty two years physician to King James 1 and Charles 1. PMID- 11395937 TI - A response to 'measles, mumps, rubella vaccine: through a glass, darkly' by Drs AJ Wakefield and SM Montgomery and published reviewers' comments. PMID- 11395938 TI - A sixty day war of words: is MMR linked to autism? PMID- 11395939 TI - MMR and toxicology. PMID- 11395940 TI - MMR safety studies. PMID- 11395941 TI - Increasing incidence of autism. PMID- 11395942 TI - The toxicity of styrene monomer. PMID- 11395943 TI - Implementing revised guidelines on euthanasia and preventing farm animal disease transmission. PMID- 11395944 TI - Cataracts and reduced fertility in degus (Octodon degus). Contracts secondary to spontaneous diabetes mellitus. PMID- 11395945 TI - Careers: people who make lab animal research happen. AB - What attracts individuals to laboratory animal research? Why do they stay? What do they enjoy about dealing with rodents and other small animals on a day-to-day basis? And why do they think it's a career others will enjoy, too? Here, we profile several lab animal professionals. PMID- 11395946 TI - Master of laboratory animal science program: teaching the science of laboratory animal science. AB - The administrators of the only program of its kind in the US explain the demand for a Master's degree in Laboratory Animal Science, the value of the degree, and plans to expand the program to make it available for distance learning. PMID- 11395947 TI - Is there a shortage of laboratory animal veterinarians? AB - There is evidence of a shortage of qualified laboratory animal veterinarians. Based on conversations with directors of animal care programs and heads of laboratory animal medicine training program, the author explores the problem of attracting veterinary school graduates to the field. PMID- 11395948 TI - An improved method for evaluating hardwood animal bedding products. AB - Technological improvements have led to the development of higher quality hardwood bedding products with lower dust content. Nevertheless, testing procedures used to evaluate the quality of laboratory animal bedding products have not kept pace, resulting in the continued use of flawed and outdated test methods. The present study was conducted to develop an improved method for evaluating the quality of hardwood animal bedding products. PMID- 11395949 TI - Hypericum perforatum: nature's mood stabilizer. AB - Hypericum perforatum (HP), better known as St. John's Wort, has been used clinically for centuries. Modern usage is still quite diverse and includes kidney and lung ailments, insomnia and depression. Standardised extracts of HP are widely used in the treatment of psychovegetative disorders and especially for mild forms of depression. Several bioactive constituents of this plant may play important role in its well-known antidepressant activity, which are discussed in the present article. Furthermore, emphasis is also given on its botany, chemistry, pharmacology and clinical efficacy. PMID- 11395950 TI - Polyamines and plant alkaloids. AB - Naturally occurring alkaloids are nitrogenous compounds that constitute the pharmacogenically active basic principles of flowering plants. Alkaloids are classified into several biogenically related groups. Tobacco alkaloids are metabolised from polyamines and diamines putrescine and cadaverine. N-methyl transferase is the first enzyme in alkaloid biosynthetic pathway which drives the flow of nitrogen away from polyamine biosynthesis to alkaloid biosynthesis. Arginine decarboxylase has been suggested to be primarily responsible for providing putrescine for nicotine synthesis. Tryptophan is the precursor of indole alkaloids. However, the biosynthetic pathway of tropane and isoquinoline alkaloids are not clear. Genes for several key biosynthetic enzymes like arginine decarboxylase, ornithine decarboxylase, putrescine N-methyl transferase and spermidine synthase, hyoscyamine 6 beta hydroxylase,tryptophan decarboxylase etc have been cloned from different plant species. These genes are regulated by plant hormones, light, different kinds of stress and elicitors like jasmonates and their strong expression is primarily in the cultured roots. In view of this, the axenic hairy root cultures induced by Agrobacterium rhizogenes have been utilised to synthesise secondary metabolites. The current development in the knowledge of alkaloid biosynthesis, particularly molecular analysis, has been discussed in this review that may help to open up new avenues of investigation for the researchers. PMID- 11395951 TI - Immunostimulatory effect of azadirachtin in Oreochromis mossambicus (Peters). AB - The effect of azadirachtin, a triterpenoid derived from Azadirachta indica on the immune response was studied in the freshwater teleost, O. mossambicus. Bovine serum albumin (BSA) and sheep erythrocytes (SRBC) were used as antigens to evoke immune response. The immune responses in fish were measured by quantifying antibodies produced and counting the peripheral blood leucocytes in control and experimental fish. In general, azadirachtin significantly enhanced the antibody response and leucocyte count in a dose dependent manner. An inverse relationship was observed between the dose of azadirachtin and the degree of immunostimulation. Timing of azadirachtin administration in relation to immunization revealed that the maximum enhancement of antibody response was observed when the stimulant was given two days prior to immunization. The observed immunostimulatory property of azadirachtin has an implication in the maintenance of finfish health in freshwater intensive aquaculture practices. PMID- 11395952 TI - Characterization of anionic amino acid transport systems in mouse mammary gland. AB - L-glutamate was transported into mammary tissue via Na(+)-dependent system XAG- that strongly interacted with both D- and L-isomers of aspartate but only with L isomer of glutamate. Replacement of Cl- by gluconate from the extracellular medium did not affect the uptake of L-glutamate. Although neutral amino acids weakly inhibited the uptake of L-glutamate, there was no evidence for the heterogeneity of anionic amino acid transport system. The XAG- system was inhibited by sulfhydryl group blocking reagent N-ethylmalemide. Low pH (6) partially inhibited the uptake by L-glutamate by mammary tissue. Prior loading of mammary tissue with L-glutamate slightly down regulated its uptake. Culturing pregnant mouse mammary tissue explants in vitro in the presence of lactogenic hormones (insulin plus cortisol plus prolactin) did not affect appreciably the uptake of L-glutamate. PMID- 11395953 TI - SDS-PAGE analysis of caput epididymis proteins in rats receiving a zinc deficient diet. AB - Caput epididymis proteins from control, pairfed and zinc deficient (ZD) wistar weanling albino rats after 2-, 4-, 6- and 8-weeks were examined using SDS-PAGE followed by densitometric scanning of the gels. In comparison to the control and pairfed rats, ZD rats displayed new proteins. These included a Mr 42 kDa from 2ZD, Mr 47.5, 27.5, 23.2 and 16.0 kDa from 4ZD and Mr 87 and 14.2 kDa from 6ZD group. The 8ZD group, however, revealed no additional protein bands over controls. Further, several other proteins were missing from ZD rats. These included Mr 93 and 71 kDa from 2ZD; 93, 90, 79, 67, 62, 55 and 15.3 kDa from 4ZD; 60, 45.5, 34, 30 and 24 kDa from 6ZD and 41.5, 33 and 27.5 kDa bands from 8ZD group. The results indicate that the induced Zn-deficient state may be responsible for the altered protein patterns in the caput epididymis. The duration of low Zn uptake period also appears to influence the protein pattern in caput epididymis. PMID- 11395954 TI - Influence of photoperiods on glycemic and adrenal catecholaminergic responses to melatonin administrations in adult male roseringed parakeets, Psittacula krameri Neumann. AB - Effects of daily (one hour prior to onset of darkness) injection of melatonin (25 micrograms/100 g body wt. for 30 days) on concentrations of blood glucose and adrenal catecholamines were studied in adult male roseringed parakeets, P. krameri under both natural (NP; about 12L:12D) and artificial long (LP; 16L:8D; lights were available in between 0600 and 2200 hrs) or short (SP; 8L:16D; lights were available between 0600 and 1400 hrs) photoperiodic conditions. The results indicate that neither LP, nor SP as such exerts any significant effect on blood glucose titre of control (vehicle of hormone administered) birds. Treatment with melatonin, however, induced hyperglycemia in both NP and LP bird groups, but hypoglycemia in SP birds. Unlike glycemic levels, amount of epinephrine (E) and norepinephrine (NE) in adrenals of control birds exhibited significant changes under altered photoperiods. A decrease in E and an increase in NE were noted in adrenals of both LP and SP birds. Exogenous melatonin in NP birds also caused a decrease in E and concomittant rise in NE levels. On the other hand, treatment of melatonin in both LP and SP bird groups resulted in an increase in the quantity of both E and NE compared to respective values in adrenals of melatonin injected NP birds. However, relative to the amount of E and NE in adrenals of placebo treated LP and SP birds, significant effect of melatonin treatment was observed only in SP birds. The results suggest that influences of exogenous melatonin on the levels of both blood glucose and adrenal catecholamines are largely modulated by short rather than long photoperiods. PMID- 11395955 TI - Alterations in collagen metabolism in heart and kidney on dexamethasone administration in rats. AB - Total collagen content in heart decreased significantly till day 8 of dexamethasone (Dex: 2.5 mg/kg week; s.c. for 2 weeks) treatment and increased on withdrawal of Dex. Acid soluble collagen content in heart decreased till day 12 of Dex treatment, reached normal level on day 16 of Dex treatment and exhibited an increase thereafter. Pepsin solubilized fraction in heart also behaved similarly as the acid soluble fraction, but reached normal level on Dex withdrawal. The total collagen content and the acid soluble collagen in kidney decreased significantly throughout treatment as well as on Dex withdrawal whereas, the pepsin solubilized collagen fraction in kidney exhibited a significant increase from day 8 of Dex treatment and the level was maintained throughout the experiment. Incorporation of 14C-proline in both, heart and kidney was found to be reduced. Electrophoretic pattern of pepsin collagen solubilized fraction of heart and kidney revealed alterations in subunit composition and its types on Dex administration and withdrawal. Thus, administration of Dex induced alterations in the metabolism of collagen and on Dex withdrawal, the system slowly tended to attain normalcy. PMID- 11395956 TI - Effect of centrally administered nitric oxide modulators in Brewer's yeast induced nociception in rats. AB - Possible modulation of Brewer's yeast-induced nociception by centrally (icv) administered nitric oxide (NO) modulators, viz., NO synthase (NOS) inhibitors, NO precursor, donors, scavengers and co-administration of NO donor (SIN-1) with NOS inhibitor (L-NAME) and NO scavenger (Hb) was investigated in rats. Administration of NOS inhibitors and NO scavenger Hb increased the pain threshold capacity significantly, whereas NO donors SIN-1, SNP and NO precursor L-arginine were found to be hyperalgesic. D-arginine, the inactive isomer of L-arginine and methylene blue, inhibitor of soluble guanylate cyclase failed to alter the nociceptive behaviour in rats. Co-administration of SIN-1 with L-NAME and Hb found to increase the nociceptive threshold. The results indicate, that centrally administered NO modulators alter the nociceptive transmission induced by Brewer's yeast in rats. PMID- 11395957 TI - In vitro schizontocidal activity of standard antimalarial drugs on chloroquine sensitive and chloroquine-resistant isolates of Plasmodium falciparum. AB - The expanding foci of multiple drug resistant malaria and emergence of different strains requires the reassessment of antimalarial activity with various drugs. In vitro response of a chloroquine sensitive and a chloroquine resistant isolate of P. falciparum to a group of 6 quinine derived and 3 artemisinin derived standard drugs has been screened, to evaluate schizontocidal activity of the drugs. In a conventional test system the IC50s were derived from the log dose response curves and evaluated by a rigorous statistical interpretation. Analysis by Tukey's test was significant for the quinine related drugs (Q < or = 0.01) and excludes the statistical significance of artemisinin related drugs in these isolates. The dose responses of these two isolates vary with quinine derivatives, with some overlap at lower doses for the sensitive isolate than for the resistant one which manifests at higher doses. PMID- 11395958 TI - Production of cephalosporin C by immobilized cells of Cephalosporium acremonium. AB - Cephalosporium acremonium ATCC 48272 cells were immobilized on various adsorbents and in various entrapment matrices. The influence of the incubation period, the best immobilization technique and the optimum concentrations of the selected matrices were investigated. From the results of the repeated batch fermentation in shake flasks, a good level of antibiotic was maintained for a period of about 19 days using 4% calcium alginate and 1% glass wool as entrapment and adsorbent supports, respectively. PMID- 11395959 TI - Paracetamol and conventional antimalarial drugs induced hepatotoxicity and its protection by methionine in rats. AB - Hepatotoxicity, induced in rats, by treatment with high doses of paracetamol and chloroquine was confirmed by estimating blood transaminase levels. Hepatoprotective effect was determined by administering combination of methionine (10% of paracetamol/chloroquine, p.o.) and hepatotoxic drugs quinine. The results were confirmed by histopathological examination of liver. Paracetamol (7 g/kg) and chloroquine (970 mg/kg) administration increased significantly the transaminase levels. Methionine alone did not produced any change. Hepatonecrosis induced by paracetamol, chloroquine alone and their combinations and its protection with methionine was revealed by histopathological study whereas the combination of paracetamol and methionine showed no significant histopathological difference when compared to the normal liver section. The results reveal that, methionine significantly prevented the rise in transaminases levels produced by hepatotoxic doses of paracetamol and chloroquine. But, to prevent occasional cases of paracetamol overdosage, it is not advisable to give methionine concurrently with paracetamol to patients who are taking paracetamol therapeutically. PMID- 11395960 TI - L-asparaginase activity in Aeromonas sp. isolated from freshwater mussel. AB - Aeromonas sp. from Lamellidens marginalis produced L-asparaginase when grown at 37 degrees C. The optimum enzyme activity was at pH 9 when temperature was 45 degrees C. Half-life of partially purified enzyme at 50 degrees C and 55 degrees C was 35 and 20 min, respectively. Activation and deactivation energies of partially purified enzyme were 17.48 and 24.86 kcal mol-1 respectively. The enzyme exhibited a Km (L-asparagine) value of 4.9 x 10(-6) mol l-1 and a Vmax of 9.803 IU ml-1. Three metal ions inhibited the enzyme activity at 10-20 mumol l-1 concentrations. Catalytic activity was also inhibited by EDTA, iodoacetic acid, parachloromercuribenzoic acid and phenylmethylsulphonyl fluoride at 0.1 mumol l 1. PMID- 11395961 TI - Colonization of arbuscular-mycorrhizal fungi on Ri T-DNA transformed roots in synthetic medium. AB - Hairy root culture of tomato (Lycopersicon esculantum L.) was induced with three strains of Agrobacterium rhizogenes namely A4, ATCC 15834 and LBA 9402. The best response in terms of growth of hairy root was observed with A. rhizogenes strain A4 and LBA 9402 followed by ATCC 15834. Hairy roots were maintained on Murashige and Skoog (MS) medium but it could also grow on minimal (M) medium. Spores of Gigaspora margarita were isolated by wet sieving and decanting method and further recovered by sucrose density gradient method. A new method for surface sterilization of spores has been described which is simpler than the methods described earlier. Surface sterilized spores of G. margarita were used for inoculation of transformed roots grown on M medium as it was found more favourable for germination and growth of spores. During co-cultivation, mycorrhizal spore germination and its penetration into root cortex were observed. Inter and intracellular mycelial spread and formation of arbuscules were also observed in the cortical region of transformed roots of this plant. PMID- 11395962 TI - Isolation and characterization of Salmonella Gallinarum cytotoxic factors. AB - Two distinct cytotoxic factors isolated from a Salmonella Gallinarum strain recovered from a bird died during an outbreak of fowl typhoid were purified to homogeneity through ciprofloxacin extraction, salt precipitation, dialysis, gelfiltration, ionexchange chromatography and chromatofocusing. These were designated as Salmonella Gallinarum cytotoxin I (GCT-I) and II (GCT-II). GCT-I was a glycoprotein having mol.wt and pI of Ca 70 kDa and 8.8, respectively. It was lethal to birds (LD50, 150 micrograms) inducing fowl typhoid like lesions. GCT-II, a protein with Ca 55 kDa mol.wt., was not lethal but caused haemorrhagic diarrhoea on intraperitoneal inoculation in birds. Both the cytotoxins induced cytopathic effects (CPE) in Vero and Madin Darby bovine kidney (MDBK) cells, enterotoxicity in rabbit ileal loop, dermatotoxicity in the rabbit skin and specific neutralizing antibodies in rabbits. These were active only between a narrow pH range of 6 to 8.5 and thermostable at 90 degrees C (1 min) but lost their activities on boiling. Trypsin and chymotrypsin enhanced their cytotoxicity, while pepsin, papain, protease, lipase and urea (5 M) had no appreciable effect on their cytotoxicity. Sodium carbonate (0.05 M) and formaldehyde (0.05%) had no effect on antigenicity of both the cytotoxic factors but rendered them nontoxic. Identification and characterization of cytotoxic moieties of S. Gallinarum not only reveals the important virulence factor but also indicates about the use of toxic factors as a candidate for toxoid vaccine and immunodiagnostics. PMID- 11395963 TI - Axenic germination of Scutellospora erythropa and Scutellospora nigra in in vitro conditions. AB - Spores of Scutellospora erythropa and Scu. nigra isolated from neem rhizosphere soils from coastal regions of Chennai were tested for axenic germination in in vitro conditions. They showed positive results in media of different composition using root exudates, soil extract, thiamine HCl and inositol. The combined medium increased the spore germination in Scu. erythropa and in Scu. nigra over water agar control. The germ tube often grew up to 3.8 cm on combined media but no vegetative spores and extramatrical auxillary cells were observed during the experiment. There was significant increase in hyphal growth when the roots were introduced into the medium, 3 days after spore germination. PMID- 11395964 TI - Rapid micropropagation of Geodorum densiflorum (Lam) Schltr. in liquid culture. AB - Technique for rapid mass propagation of Geodorum densiflorum (Lam)Schltr. has been developed by using thin sections of stems of in vitro regenerated plantlets as explant source. Thin sections of stems (1.0-1.5 mm) when cultured in modified liquid and semisolid Knudson C (KnC) medium produced 1.8 and 1.2 protocorm like bodies (PLBs) per explant respectively. Peptone (2 g l-1) was effective in promoting the survival percentage of the explants but had no effect on PLB production. BAP and NAA when used individually enhanced the rate of PLB production. But a significant and manifold increase in PLB production was noted when BAP (3 mg l-1) and NAA (0.5 mg l-1) in combination were added to peptone supplemented liquid and semisolid KnC medium. PLBs thus obtained were subcultured in semisolid KnC medium and obtained well developed plantlets within 10-12 weeks. PMID- 11395965 TI - Biodiversity of Anabaena azollae isolates from different Azolla cultures. AB - The random amplified polymorphic DNA (RAPD) profile of A. azollae strains isolated from four different Azolla cultures was studied by using different primers. The objective of this study was to determine whether polymerase chain reaction (PCR) with different primers could differentiate the isolated A. azollae strains from one another. The primers amplified specific sequences of the isolates and generated fingerprinting pattern characteristic of each isolate. Clear polymorphism was noticed among all the strains which depends on the primer sequence. PMID- 11395966 TI - Role of ATP sensitive potassium channel on 7-hydroxy flavone induced antinociception and possible association with changes in glycaemic status. AB - Opioid type of analgesics open ATP sensitive potassium channel at the cellular level to produce antinociceptive response. These channels have also been shown to modulate insulin secretion by the pancreas. 7-hydroxy flavone, an antinociceptive agent shown to act through opioid pathways was investigated for its effect on glycaemic state and associated algesic state. The involvement of ATP sensitive potassium channel in the action was examined by using glybenclamide. The result reveal that 7-HF per se did not elicit any significant change in the glycaemic state simultaneously eliciting antinociceptive response as tested by acetic acid induced abdominal constriction assay procedure. Glibenclamide treatment attenuated the antinociceptive effect of 7-HF and while maintained its hypoglycaemic response. The present finding suggest that 7-HF induces antinociception like morphine, utilise ATP sensitive potassium channel at the cellular level and do not suggest a cause-effect relationship between the changes in the glycaemic and algesic state. Possibly, insulin which is controlled by ATP sensitive potassium channel at the cellular level might also modulate antinociception exhibiting a cause-effect relationship between them. PMID- 11395967 TI - Enhancing the efficacy and persistency of Spodoptera litura (Fab.) nuclear polyhedrosis virus using UV irradiation protectants. AB - To enhance the field persistency of S. litura nuclear polyhedrosis virus (SLNPV), three chemicals viz. cupric ammonium nitrate, tinopal and cupric sulphate were tried as protectants (0.01 mg/ml) against natural sunlight (UV) irradiation. On exposure for 8 hr and subsequent bioassaying (diet surface treatment), it was found that cupric sulphate protected the polyhedrosis inclusion bodies (PIBs), recording 95.56% mortality which was statistically at par with unexposed PIBs recording 97.78% mortality. PMID- 11395968 TI - Anti-B cell and anti-cytokine therapy for the treatment of post-transplant lymphoproliferative disorder: past, present, and future. AB - Epstein-Barr virus (EBV)-induced post-transplant lymphoproliferative disorder (PTLD) is a rare but severe complication of solid organ and bone-marrow transplantation. When possible, reduction of immunosuppressive treatment or surgery for localized disease may cure PTLD. Therapeutic approaches using chemotherapy or antiviral drugs have limited effects on survival. Adoptive immunotherapy with donor T cell infusions has shown promising results in bone marrow transplantation. The use of anti-B cell monoclonal antibodies is herein reported as a safe and efficient therapy for severe PTLD. Because IL6 has been described as a growth factor for EBV-infected B cells, anti-IL6 monoclonal antibody therapy was also tested in a phase I-II clinical trial, the results of which are summarized. PMID- 11395969 TI - Interferon and cytotoxic chemotherapy for the treatment of post-transplant lymphoproliferative disorder. AB - Interferon-alpha and cytotoxic chemotherapy may be effective treatment modalities for the post-transplant lymphoproliferative disorder. Interferon-alpha may result in a complete response in up to 40% of patients, while chemotherapy may be effective in 75% of those failing local surgical excision, a reduction in immunosuppression, and an antiviral agent. Interferon may be used early after diagnosis in patients with relatively slowly growing tumors. Chemotherapy should be selected for patients with bulky, rapidly growing malignancies. The toxicity of chemotherapy may be minimized by discontinuing maintenance immunosuppression during chemotherapy, administering GCSF, and providing antimicrobial prophylaxis. Rejection is minimized by the reintroduction of maintenance immunosuppression when the patient is no longer neutropenic. PMID- 11395970 TI - Epstein-Barr virus infection and malignancy in solid organ transplant recipients: strategies for prevention and treatment. PMID- 11395971 TI - The Epstein-Barr virus and post-transplant lymphoproliferative disease: interplay of immunosuppression, EBV, and the immune system in disease pathogenesis. AB - Transplant patients are at particular risk for developing post-transplant lymphoproliferative disease (PTLD) following administration of immunosuppressive therapy. In many cases the PTLD lesions express Epstein-Barr virus (EBV) latent and lytic genes as well as elevated levels of host cytokines. An outline of the potential contributions of EBV, host cytokines and T cells, and the immunosuppressive cyclosporine A, tacrolimus, and anti-CD3 antibody in the mechanism and pathogenesis of this disease is presented and discussed. PMID- 11395972 TI - Identifying the patient at risk for post-transplant lymphoproliferative disorder. AB - Post-transplant lymphoproliferative disorders (PTLD) are a recognized complication of the immunosuppression required to prevent allograft rejection, occurring in 1-20% of recipients of solid organ transplants. Several factors greatly increase the risk of developing PTLD early post-transplant in any individual recipient. Epstein-Barr virus (EBV) infection is critical in the pathogenesis of the majority of these cases. Pre-transplant EBV seronegativity increases the incidence of PTLD 10- to 75-fold over that of EBV-seropositive recipients. Other risk factors include very young recipient age, cytomegalovirus infection or mismatching (donor positive-recipient negative), aggressive immunosuppression with conventional biologic agents, and the type of organ transplanted. In contrast, the risk of developing PTLD late in the post transplant course does not appear to be influenced by the type of immunosuppressive agents employed, but rather by the duration of any immunosuppression. The role of EBV in late PTLD is also less certain, as a greater proportion of lesions are not associated with evidence of EBV infection. As the understanding of these risk factors has expanded, opportunities exist to target those populations at highest risk for the development of PTLD for aggressive monitoring and pre-emptive or prophylactic therapy. It is hoped that implementation of such strategies will render early PTLD a preventable complication of transplantation. PMID- 11395973 TI - Epstein-Barr virus load monitoring: its role in the prevention and management of post-transplant lymphoproliferative disease. AB - The Epstein-Barr virus load in the peripheral blood at the time of diagnosis of post-transplant lymphoproliferative disease (PTLD) is elevated 1000- to 10,000 fold compared to the level detected in normal latency. With the use of quantitative polymerase chain reaction (PCR), changes in the viral load over time can be measured with a two- to fourfold accuracy. This has allowed early detection of first-time infections and reactivations that may lead to PTLD and has provided an opportunity to intervene before symptomatic disease has occurred. Viral load monitoring has also been used to follow patients with PTLD and, along with other parameters, provided an assessment of the effectiveness of therapeutic protocols. Viral load monitoring has led to the discovery that at least two thirds of transplant recipients become persistent viral load carriers. While the persistent load appears to be largely carried in latently infected memory B cells, more work is needed to clearly define this type of persistent infection and determine the risks associated with it. New diagnostic tests need to be developed to distinguish the persistent latent viral loads from viral loads that are likely to become symptomatic PTLD. PMID- 11395974 TI - The diverse pathology of post-transplant lymphoproliferative disorders: the importance of a standardized approach. AB - Post-transplant lymphoproliferative disorders (PTLD) are a diverse group of abnormal lymphoid growths that include both hyperplasias and neoplasias. They have been divided into several general pathologic categories that have prognostic significance. These include early or hyperplastic PTLD, polymorphic PTLD, and lymphomatous or monomorphic PTLD. The majority of PTLDs are of B-cell origin and contain Epstein-Barr virus (EBV). However, PTLDs of T- or NK-cell origin have been described, and late-arising EBV-negative lymphoid tumors are becoming more frequently reported in this population. Other lymphoid neoplasms, such as those arising from mucosal-associated lymphoid tissue (MALTomas), have recently been recognized in transplant patients, and their relationship to PTLD is uncertain. Multicentric PTLD may represent either advanced-stage disease or multiple independent primary tumors. Likewise, recurrent PTLD may represent true recurrence or the emergence of a second primary tumor. Transplant patients are also at risk for other opportunistic neoplasms, including EBV-associated leiomyosarcomas that may be seen alone or in conjunction with PTLD. This underscores the necessity for pathologic diagnosis of mass lesions in this patient population. The pathologist should strive to categorize the form of post transplant lymphoproliferation in accordance with currently accepted criteria. The diagnosis should incorporate the histopathologic appearance, cell phenotype, clonal status, and EB viral status. The pathologist may play a special role in guiding therapy by ascertaining the presence of such markers as CD20 on tumor cells. Specialized techniques, such as molecular analysis of oncogenes/tumor suppressor genes and evaluation of host:donor status of PTLD, may play important roles in diagnostic evaluation in the future. PMID- 11395975 TI - The role of antiviral and immunoglobulin therapy in the prevention of Epstein Barr virus infection and post-transplant lymphoproliferative disease following solid organ transplantation. AB - The recognition of the importance of Epstein-Barr virus (EBV) infection, including EBV-associated post-transplant lymphoproliferative disease (PTLD), has led to a new focus on the prevention of this problem. This paper reviews the scientific rationale behind, and clinical experience with, the use of chemoprophylaxis (using acyclovir or ganciclovir) and immunoprophylaxis (using intravenous immunoglobulin) in the prevention of EBV/PTLD. While some centers have already introduced the use of one or both of these agents as standard prophylaxis against the development of this complication, published data in support of these protocols are currently lacking. Well designed clinical trials are necessary to evaluate the potential role of both antiviral and immunoglobulin agents in the prevention of EBV/PTLD in organ transplant recipients. PMID- 11395976 TI - Classifying medical devices according to their maintenance sensitivity: a practical, risk-based approach to PM program management. AB - Many medical equipment items need periodic attention to ensure that they continue to operate properly and safely; and most inspecting agencies require healthcare facilities to have a competent equipment maintenance program that is focused on the most critical of those devices. There is however a long-standing lack of consensus on how best to determine which devices should be included in this critical device category, and which can reasonably be excluded. A new methodology is proposed for establishing a logical, fact-based framework for determining which devices should be included. It is based in part on a new FDA-sanctioned definition of what an appropriate regimen of planned maintenance activities for a medical device should include. This new definition addresses the medical device users' concerns about periodic performance verification and safety testing as well as detecting and correcting the wear or progressive deterioration of any non durable parts, which is the primary focus of the conventional preventive maintenance programs found in less critical industries. The analytical approach proposed utilizes technical information that is either already available or which can be easily developed. It characterizes each different device by means of a 3 letter maintenance sensitivity profile that can be used to analyze the effectiveness of the maintenance procedures as well as quantify the device's sensitivity to planned maintenance. A collaborative effort to assemble and organize this data would provide the industry with a sound, logical platform for narrowing the scope of most PM programs and allow us to redirect a significant amount of scarce technical manpower into more productive activities such as device user training. PMID- 11395977 TI - The strategic impact of information technology convergence on healthcare delivery and support organizations. PMID- 11395978 TI - Report of the American Medical Association (AMA) Council on Scientific Affairs and AMA recommendations to medical professional staff on the use of wireless radio-frequency equipment in hospitals. AB - In lieu of a comprehensive technical assessment, this report of the American Medical Association (AMA) Council on Scientific Affairs reviews the issue of electromagnetic interference (EMI) with medical devices caused by the use of wireless radio-frequency (RF) equipment such as cellular telephones, 2-way radios, and pagers. It identifies medical device EMI problems that have been reported and guidance that has been published to assist hospital personnel in assessing and minimizing the potential for EMI in the hospital and in making informed risk management decisions. For more-detailed information on this subject, readers are encouraged to consult publications from various scientific and health care organizations. PMID- 11395980 TI - What kind of information technology training do you need and where can you find it? PMID- 11395979 TI - The way to create a competitive difference in a commodity world. PMID- 11395981 TI - A repeater and bridge primer. PMID- 11395982 TI - Protein hydrolysate vs free amino acid-based diets on the nutritional recovery of the starved rat. AB - BACKGROUND AND AIMS: To test the hypothesis that a peptide-based enteral product was equivalent to a low-fat, free amino acid-based formula in the nutritional and functional recovery of the starved rat. METHODS: Sixteen male Wistar rats were starved for 3 days. Then, rats were randomised to a whey protein hydrolysate based diet or a free amino acid-based diet and refed for 3 days. The experiment was designed to provide the same energy intake in both groups. The parameters studied included body weight gain, nitrogen retention, plasma free amino acid concentrations, muscle glutamine concentrations and glutathione levels in gut mucosa and liver. RESULTS: Weight gain was statistically higher on the peptide based diet than on the elemental diet after the refeeding period. This difference in weight gain was associated with a statistically higher nitrogen retention. Plasma and muscle free glutamine concentrations were higher in rats fed the whey protein hydrolysate-based diet than those in rats refed the free amino acid-based diet, even though the glutamine intake was higher in the latter group. Glutathione concentrations in liver and gut mucosa were similar in the groups. CONCLUSION: We conclude that enteral diets containing peptides were more effective than a diet containing free amino acids in the nutritional recovery of the starved rat. PMID- 11395983 TI - Effect of diet on the replenishment of intramyocellular lipids after exercise. AB - BACKGROUND: Muscle triglycerides are important as a source of energy and in relation to metabolic sensitivity. However, the classic biopsy method does not distinguish intra- from extracellular fat, and their regulation by exercise and diet is largely unknown. Magnetic resonance spectroscopy (MRS) is available to assess the intramyocellular lipid (IMCL) pool non-invasively in humans. AIM OF THE STUDY: The aim of this work was to use sequential MRS measurements of IMCL and glycogen to explore the role of three levels of dietary fat on the replenishment of these energy stores after exercise. METHODS: Following 2 h of exercise, two subjects (S1, S2) were fed one of three diets (15%, 40% or 70% fat energy), each on a separate occasion. IMCL and glycogen were measured by MRS in the tibialis anterior muscle before, after exercise, and at 10 and at 32 h of recovery. RESULTS: Initial IMCL concentration (mmol.kg-1: 3.0 in S1 and 1.8 in S2) was reduced to 70% after exercise. The rate of replenishment was minimal with the low-fat (mmol.kg-1.24 h-1: 0.7 and 0.0) and much higher with both higher fat diets (mmol.kg-1.24 h-1: 3.1 and 3.2 in S1, 0.7 and 0.9 in S2). Glycogen and IMCL replenishments were inversely correlated. CONCLUSIONS: IMCL and glycogen can vary acutely in response to diet after exercise. Studies are needed to determine if such variations occur within the range of ordinary diets and to clarify the functional significance of IMCL in differently active individuals. PMID- 11395984 TI - Total artificial nutrition is associated with major changes in the fecal flora. AB - BACKGROUND: Animal studies have demonstrated dramatic changes in the intestinal flora during total enteral (TEN) or parenteral (TPN) nutrition. AIM OF THE STUDY: To assess the impact of TEN and TPN on human intestinal microflora. METHODS: Eight patients on fiber-free TEN, five patients on TPN, and ten controls were studied. Fecal bacteria were identified and numbered (logCFU/g feces), and fecal short-chain fatty acids (SCFAs) were measured in stool samples, by gas-liquid chromatography. RESULTS: In TEN patients, compared to controls (P < 0.01), aerobes were increased (8.46 +/- 0.24) while anaerobes were decreased (5.79 +/- 0.84). In TPN patients, both aerobes and anaerobes were decreased compared to controls (5.64 +/- 0.27 and 5.31 +/- 1.09 respectively, P < 0.01). Total SCFAs were lower in TPN patients than in TEN patients (48.3 +/- 16.6 vs 118.6 +/- 24.1 mmol/kg, P < 0.05). CONCLUSIONS: Both TPN and TEN induce modifications in the intestinal microflora. During TPN, a homogeneous decrease occurs in both aerobic and anaerobic bacteria. TEN decreases only anaerobic bacteria, while aerobic bacteria are increased. This imbalance may play a role in the pathophysiology of TEN-induced diarrhea. PMID- 11395986 TI - School-administered weekly iron supplementation--effect on the growth and hemoglobin status of non-anemic Bolivian school-age children. A randomized placebo-controlled trial. AB - BACKGROUND: Recent data suggest that daily iron supplementation of iron-replete children could impair their growth. If verified for weekly iron supplementation these results would markedly complicate targeting and implementing school-based weekly iron supplementation programs. AIM OF THE STUDY: To ascertain the effect of weekly iron supplementation on the growth and hemoglobin status of non-anemic school-age children. SUBJECTS AND METHODS: 73 Bolivian non-anemic school-age children randomly assigned to the treatment group (n = 37; receiving supplements containing FeSO4 during 18 weeks) or the control group (n = 36; receiving a placebo during the same period). Hemoglobin concentration and anthropometric measures were determined for each child at the beginning (T0) and the end (T18) of the study. RESULTS: The treatment group did not show any significant variation in hemoglobin concentration between T0 and T18 (-1.6 +/- 10.4 g/L; P = 0.40) whereas the control group showed a significant decrease in hemoglobin concentration (-4.6 +/- 10.9 g/L; P = 0.03). Anthropometric changes were not significantly different between the treatment and the control groups for weight, (1.63 +/- 1.11 kg vs 1.88 +/- 0.79 kg; P = 0.30), height (2.35 +/- 0.94 cm vs 2.11 +/- 1.03 cm; P = 0.34) or mid-upper arm circumference (0.29 +/- 0.57 cm vs 0.22 +/- 0.54 cm; P = 0.64). CONCLUSION: In our study, weekly iron supplementation of non-anemic school-age children had no negative effect on their growth while having a positive effect in preventing significant decreases in hemoglobin concentration. These results suggest that in regions where iron deficiency anemia (IDA) is prevalent, a simple and cost-effective way to control IDA in school-age children is to give weekly iron supplements to all children at school. PMID- 11395985 TI - Effects of diary food supplements on bone mineral density in teenage girls. AB - BACKGROUND: Bone mineral density (BMD) is largely genetically determined and this influence is most powerful in the period of rapid skeletal development in childhood and late adolescence but environmental factors such as exercise and dietary calcium intake may influence up to 20%. AIMS OF THE STUDY: The aims of the study were to examine healthy late adolescent females for the effects and benefits of a high calcium intake from dairy product foods on bone mineral density, body composition, lipids and biochemistry. The secondary aim is determine whether a high intake of dairy product foods in the diet is acceptable for this age group long term. METHODS: Ninety-one teenage girls who participated in a two-year randomised controlled study on the effect of dairy food supplementation on dietary patterns, body composition and bone density in post pubertal teenage girls were approached one year after the cessation of the study to determine the effects of the cessation of dairy supplements on bone mineral density, dietary habits, biochemical markers, body composition and blood lipids. Bone mineral density and bone mineral content were assessed at the hip, spine and total body. Anthropometric data were collected, and exercise, Tanner, dietary assessment, preference and compliance questionnaires were administered. Lipid profiles, hydroxyproline excretion and urinary calcium and sodium excretion measurements were performed. RESULTS: There were no significant differences between the 2 groups for height, weight, lean and fat mass. The supplemented group had significantly higher calcium, phosphorus and protein intake during the supplementation period (p < 0.001). No differences were seen between the groups 12 months after supplementation finished. There were no significant differences in exercise level, preference or acceptability of dairy products or in the lipids and bone markers between baseline the end of supplementation and 1 year follow up. There was a significant increase in trochanter (4.6%), lumbar spine (1.5%) and femoral neck (4.8%) BMD (p < 0.05) in the high calcium group at the end of supplementation. There was an increase in bone mineral content at the trochanter (p < 0.05) and lumbar spine; however the latter was not statistically significant, in the high calcium group at the end of supplementation. There was no difference in vertebral height or width at any stage of the study, indicating no influence on bone size. CONCLUSIONS: In this 3 year study (2 years of supplementation, 1 year follow-up), teenage girls, aged 15-18 years, were able to significantly increase their BMD at the trochanter, femoral neck and lumbar spine when supplemented with dairy product foods to a mean calcium intake of 1160 mg/d. There was also an effect seen on the BMC particularly at the trochanter and to a lesser extent at the lumbar spine. The dietary calcium intake achieved did not adversely affect body weight, fat and lean mass or blood lipid profiles. Twelve months after the supplementation finished the girls had returned to their baseline diet, indicating self-selection of a high dairy product diet may be hard to achieve. PMID- 11395987 TI - Adaptation to chronic ethanol administration emphasized by fatty acid hydroxylations in rat liver and kidney microsomes. AB - BACKGROUND: Long-term ethanol consumption in laboratory animals is associated with histological alterations of liver cells and modifications of fatty acid metabolism. AIM OF THE STUDY: The present study was aimed at investigating the effect of 1- and 2-month chronic treatment of rats with ethanol on the metabolism of two unsaturated (oleic and linoleic) fatty acids in liver and kidney microsomes, in relation to the CYP2E1 enzyme content in both tissues. METHODS: Rats were fed ethanol (14 g/Kg/d) or dextrose through a permanently implanted gastric cannula, as described in the intragastric feeding rat model for alcoholic liver disease (ALD). CYP2E1 level was immuno-quantified in both liver and kidney microsomes by Western blot, whereas fatty acid omega- and (omega-1) hydroxylations were measured using HPLC and radiometric analytical methods. RESULTS: One- and two-month ethanol treatment led to a 3- to 4-fold rise of the CYP2E1 protein in both liver and kidney microsomes. Oleic and linoleic acid (omega-1)-hydroxylations were increased (approximately 3-fold) in liver microsomes after one-month of ethanol administration, but surprisingly such a rise was not observed after a two-month treatment; on the other hand, no effect was observed on the omega-hydroxylations of these fatty acids. Furthermore, as previously described for lauric acid, ethanol intake did not significantly act on the kidney microsome capability to hydroxylate unsaturated fatty acids. CONCLUSIONS: CYP2E1 is strongly inducible by ethanol and therefore accounts for the tolerance for this hepatotoxicant. Our results support the development of an adaptation process in the liver hydroxylating enzyme system, which occurs between one and two months of ethanol feeding. Although it is usually not appropriate to extrapolate animal findings to humans, rat and human CYP2E1s were observed to have comparable specificities and similar mechanisms of regulation. Thus, the present study allowed the acquirement of detailed information of CYP2E1 activity in patients with severe manifestations of ALD. PMID- 11395988 TI - [Interventional therapy after failed fibrinolysis in acute myocardial infarct. Acute and long-term outcome of referral for rescue balloon angioplasty]. AB - BACKGROUND: The results from studies of coronary angioplasty after failed thrombolysis (rescue-PTCA) in acute myocardial infarction are contradictory. Long term results were not presented till now. Therefore we analyzed the data from our registry of those patients whose acute and long-term results were available. PATIENTS AND METHODS: Data of 49 patients were analyzed who had been admitted for rescue-PTCA from other hospitals. Thrombolysis had to be started < 6 hours (mean 2.7 hours) from onset of symptoms. Rescue-PTCA had to be completed within < 24 hours (mean 10.5 hours). 37 patients received streptokinase, seven rt-PA, three urokinase and two prourokinase. Electrocardiographic and clinical criteria were used to define failure of thrombolysis. The data of the acute results were from a prospective registry and the long-term results came from clinical follow-up visits and a questionnaire sent to the patients. RESULTS: Mean age of the patients was 48.5 years (38-78 years), 45 male, nine patients in cardiogenic shock (18%), infarct related artery (IRA): RCA 22x, LAD 21x, LCX 5x, CABG 1x, single vessel disease 27x, multiple vessel disease 22x. Acute results: Initial IRA-TIMI flow 0 in 28 patients, 1 in twelve patients, 2 in 9 patients; after rescue-PTCA TIMI flow 1 in one patient, 2 in two patients, 3 in 46 patients (procedural success 94%). Hospital mortality 8.2% (four patients), all in cardiogenic shock. Early reocclusion rate 10%. Bleeding complications 14%, no fatal complications. Long-term results: Observation period 2.5 years in 42 patients (0.5-6.5 years). Three more deaths. Total mortality 14% (7/49). Angiographic follow-up: Ejection fraction initially 50%; 53% after 3 months. Repeat revascularization in 43% (15/35): Re-PTCA in 8/35, surgery in 6/35 patients, 1x transplantation. 80% of the patients were free from angina or heart failure. CONCLUSIONS: Rescue-PTCA in acute myocardial infarction has a high procedural success rate with a low hospital mortality. It is the treatment of choice for patients in cardiogenic shock. Transportation to an interventional center is safe. The reintervention rate is comparably high. The long-term results are good. PMID- 11395989 TI - [Myocardial ischemia in hypertensive patients]. AB - AIM: To examine the frequency of myocardial ischemia in hypertensive patients with insufficiently adjusted blood pressure. It was to clarify the question if there is a difference between hypertensives with silent ischemic episodes alone and hypertensives with symptomatic episodes as well, referring to the degree of severity of ischemia. A resulting question was: if there is a difference between them, might it be possible to influence this by lowering the blood pressure? PATIENTS AND METHODS: 104 hypertensive patients had a check-up before and after the suspension of normotensive blood pressure by means of a 24-hour long-term ECG. Significant results for a myocardial ischemia were horizontal and descending ST segment depression > or = 0.1 mV for the duration > or = 1 min. RESULTS: 38 of the examined hypertensives showed 316 silent and 19 symptomatic episodes of myocardial ischemia. Hypertensives with additionally symptomatic episodes (n = 8) showed a severity of ischemia (ST-total area) which was significantly higher than in patients with only silent episodes (9.3 [2.7-65.3] mV min vs 2.6 [0.1-42.6] mV min, p < 0.05). After reduction of the blood pressure these patients as well as patients with silent episodes a lone (2.6 [0.1-42.6] mV min vs 0 [0 to 39.3] mV min, p < 0.0001) were found to have a decline of the ST-total area (9.3 [2.7 65.3] mV min vs. 0 [0-46.5] mV min, p < 0.05). There was no significant difference between them any longer. CONCLUSION: The severity of ischemia in insufficiently adjusted hypertensive patients with additionally symptomatic myocardial ischemia is higher than in the case of patients with silent episodes alone. After reaching a normotensive blood pressure again a difference was no longer evident. PMID- 11395990 TI - [New concepts of primary prevention require rethinking]. AB - The decline in cardiovascular mortality over the past decades is due to improved survival of patients with clinical events rather than to a declining incidence of these events. As a result, an increased prevalence of chronic coronary artery disease and especially congestive heart failure has been described. Further substantial reductions in coronary artery disease morbidity and mortality can only be anticipated if coronary artery disease is treated before the manifestation of clinical disease. Modern concepts of primary prevention incorporate an individualized approach to risk assessment. Medical history, physical examination, and established laboratory tests are the basic instruments which allow for quantitative risk assessment. Tables and simplified algorithms derived from large clinical trials enable the calculation of intermediate and long-term ("life-time") probability of cardiac events. In this setting of quantitative risk assessment, the importance of some risk factors is increasingly recognized, namely diabetes. Noninvasive diagnostic testing is used to detect preclinical atherosclerotic plaque disease. Direct imaging of coronary and peripheral arteries, the ankle-brachial index, and measurements of C-reactive protein represent novel methods which have become available for further risk stratification. Because they consider the pathophysiology of atherosclerosis and coronary artery disease, these methods may facilitate decision-making regarding preventive treatment in patients who have an intermediate risk on the basis of the traditional risk factors. The modern concept of primary prevention is aimed at identifying subjects whose risk is similarly high as that of patients with clinically established cardiovascular disease. These subjects can then be treated efficiently, and the strict distinction between primary and secondary prevention is blurred. PMID- 11395991 TI - [Therapy of osteoporosis from the viewpoint of evidence-based medicine]. AB - DEFINITION AND SOCIOECONOMIC ASPECTS: Osteoporosis is a disease characterized by low bone mass and an increased susceptibility to fractures. It represents an enormous burden for the social security systems in developed countries. In Germany, approximately two million women and 800,000 men suffer from vertebral fractures and estimates for hip fracture incidence are in the range of 70,000 130,000 per year. The resulting costs for hip fractures alone could be calculated to 3-5 billion German marks. THERAPY ACCORDING TO EVIDENCE-BASED MEDICINE (EBM): According to Sackett et al. 1996, evidence-based medicine is the conscientious, explicit, and judicious use of current best evidence in making decisions about the care of individual patients. The practice of evidence-based medicine means integrating individual clinical expertise with the best available external evidence from systematic research. OSTEOPOROSIS THERAPY: The goal of osteoporosis therapy is to prevent fractures and several therapeutic options are available for this disease. With respect to proven fracture benefit, however, the quality of evidence from randomized clinical trials varies substantially among therapies. From systematic research the best external evidence is available for a supplementation with calcium and vitamin D and a therapy with the bisphosphonates alendronate or risedronate, as well as the SERM raloxifene. For other therapeutic agents like fluorides, vitamin D metabolites, calcitonin, and etidronate the quality of evidence is much lower. So far, there is no evidence for other pharmaceutical therapies. Hip protectors are effective for the prevention of hip fractures. PMID- 11395992 TI - [Anticoagulation in primary prevention of thromboembolism in advanced left ventricular dysfunction]. AB - BACKGROUND: Patients with heart failure have an increased risk for thromboembolic events. In clinical practice the physician is often confronted with the decision to establish a prophylactic anticoagulation. DATA: The incidence for clinical embolization is 1.5 to 3.5% per year. It seems that patients with a lower peak oxygen uptake and with a lower ejection fraction are at higher risk for embolic events. There is no evidence for a correlation of such events with the clinical classification (NYHA) or with the genesis of heart failure. Concerning a prophylactic anticoagulation, the results of the published studies and meta analyses are inhomogenous with a benefit in some, no difference in others and a significant disadvantage for the patients with anticoagulation in a part of them compared to a non-treated control group. None of these data is established by a prospective, randomized, controlled study with the primary endpoint thromboembolic event in patients with or without anticoagulation. The incidence for a hemorrhage under anticoagulation is 0.6-5.3% per year for life-threatening and 0.04-0.64% for fatal bleeding. CONCLUSION: Because of similar incidences for thromboembolic events and for the bleeding complication of anticoagulation, there is no evidence for a general indication for anticoagulation of patients with heart failure, persistent sinus rhythm and lack of risk factors. In patients with high-grade heart failure (e.g. VO2max < 14 ml/min/kg body weight or left ventricular ejection fraction < 20-30%), who belong to a high-risk population, anticoagulation should be considered and may be acceptable in individual cases. PMID- 11395993 TI - [Reactive hemophagocytosis syndrome as a rare cause of fatal pancytopenia]. AB - CASE REPORT: We present the case of a feverish 36-year-old female patient with unspecific pain and weight loss. After initial diagnosis of an unclear infection with Candida esophagitis, unspecific duodenitis, proctitis and liver damage there was a further clinical deterioration during antibiotic and symptomatic therapy. A newly developed pancytopenia could be identified as caused by reactive hemophagocytosis using bone marrow analysis. The patient died despite maximal supportive and specific therapy. Cause of death was a retroperitoneal hematoma and an invasive aspergillosis, the latter not identified premortem. CONCLUSION: The reactive infect-associated hemophagocytosis is a rare differential diagnosis in feverish pancytopenia. The prognosis is poor. In absence of a proven causal therapy supportive therapy and maximal diagnostics for detection of curable underlying diseases are essential. Antimycotic therapy should be considered generally. PMID- 11395994 TI - [60-year-old patient with fever, jaundice and intra-abdominal lymphomas]. AB - BACKGROUND: Infectious mononucleosis usually manifests in adolescents and young adults. Medical history in elderly patients is often atypical and severe. CASE REPORT: We report on the medical history of a 60-year-old woman, who came into our hospital with fever, icterus, decreasing performance status and abdominal pain. Splenomegaly and multiple abdominal lymph nodes could be found by ultrasound and CT scan. Endoscopically severe mucosal alterations could be found in the upper and lower gastrointestinal tract. Serologically, an acute EBV infection was diagnosed. With symptomatic treatment, the clinical course was without any problems. CONCLUSION: Casuistically, we described the atypical medical history of an elderly patient with infectious mononucleosis. In each undefined lymphadenopathy of elderly patients, an EBV infection should be included into the differential diagnosis. PMID- 11395995 TI - [Nitrate headache in blasting work]. AB - CASE REPORT: A 34-year-old white male was executing blasting works in an operation of a quarry several times a month during a period of 1 year. Each time he worked with explosives, he complained of headache, vertigo and concentration failure which started shortly after beginning and continued several hours after. The results of the physical examination on a day without blasting works were normal. Because of the exact working parallel appearance of the symptoms we did further explorations at the workplace which showed the usage of nitroglycerin blasting agents without observance of the safety regulations. The symptoms only at blasting workdays with normal clinical findings at break led considering the relation with the occupational conditions to the diagnosis "headache due to exposure to nitroglycerin explosives". It was reported to the employer's liability insurance association as an occupational disease with the aim of improving occupational conditions. CONCLUSION: Occupational toxic agents as a potential trigger of unclear headache support the requirement of an exact anamnesis of working conditions and environment. PMID- 11395996 TI - [Recurrent gluteal injection abscesses as a complication of frequent intramuscular self-injection]. PMID- 11395997 TI - [The evidence (in) the evidence-based medicine]. AB - Evidence-based medicine is a new approach to improve the transfer process of knowledge from research to medical practice. The assumption that only results of randomized controlled studies are evident is true for many but by far not for all clinical problems. As is demonstrated from one historical and many recent examples, there exists another but equally stringent method of proof which is based on an implicit historical comparison. This kind of evidence still has to be defined exactly in order to protect it from misuse by alternative medicine. The statement that only 20% of methods used in conventional medicine are [corrected] evidence-based cannot be substantiated. Methods and importance of meta-analyses are critically discussed as well as the meaning of the term publication bias. The new documentation- and information techniques will improve some steps in the transfer process. At the end it will be crucial whether the last step, the improvement of rationality in patient care, will be successful. This evidence still has to be demonstrated. PMID- 11395998 TI - [Sneddon syndrome: vasculitis or thrombotic vasculopathy? Comment on the contribution by Zipper et al]. PMID- 11396000 TI - Analysis of protein transglutamylation in apoptosis. PMID- 11395999 TI - Isolation and assay of caspases. PMID- 11396001 TI - Analysis of sphingomyelin and ceramide levels and the enzymes regulating their metabolism in response to cell stress. PMID- 11396002 TI - Cell-free systems to study apoptosis. PMID- 11396003 TI - Role of c-Jun N-terminal kinase in apoptosis. PMID- 11396004 TI - Methods for studying pro- and antiapoptotic genes in nonimmortal cells. PMID- 11396006 TI - Proteinase families and their inhibitors. PMID- 11396005 TI - Calcium flux measurements in apoptosis. AB - Early studies in apoptosis implicated an increase in cytosolic Ca2+ as a direct mediator of DNA fragmentation. However, efforts to delineate targets for this increase in Ca2+ have been slow in evolving. Several previous studies have implicated ER Ca2+ pool depletion in the initiation of apoptosis. Our own preliminary studies confirm that many (but not all) apoptotic stimuli empty the ER store via a mechanism that is blocked by BCL-2 expression. Furthermore, ER pool depletion is not affected by broad spectrum caspase inhibitors, indicating that it occurs via a caspase-independent mechanism. Finally, our data demonstrate that ER pool depletion occurs prior to release of cytochrome c from mitochondria. Given previous work demonstrating close coordination of ER and mitochondrial Ca2+ levels, we speculate that ER-dependent changes in mitochondrial Ca2+ serve as important signals for cytochrome c release. Alternative mechanisms include activation of caspase-12 and/or the JNK pathway, both of which can be directly stimulated by depletion of the ER Ca2+ pool. Although substantial improvements in intracellular Ca2+ imaging have emerged, compelling answers to many of the present questions related to the role of Ca2+ in apoptosis await future technical improvements. The development of organelle-specific, recombinant Ca2+ probes (targeted aequorins and cameleons) certainly should facilitate some of this work, although the target cell of interest must be amenable to molecular manipulation (transfection), which precludes straightforward analysis of primary cells. Pharmacological tools (i.e., thapsigargin and DBHQ) can provide conclusive data on ER pool status without requiring an overly sophisticated image analysis system. However, confocal microscopy allows for the effective analysis of Ca2+ pools as long as dye localization is homogeneous and properly controlled. However, current techniques should be considered semiquantitative at best and will remain so until specific organelle-targeted fluorescent dyes are developed and widely available. PMID- 11396007 TI - Identification and analysis of caspase substrates: proteolytic cleavage of poly(ADP-ribose)polymerase and DNA fragmentation factor 45. PMID- 11396008 TI - Cloning and analysis of Bcl-2 family genes. PMID- 11396009 TI - Analysis of reactive oxygen species in cell death. PMID- 11396010 TI - Methods for studying apoptosis and phagocytosis of apoptotic cells in Drosophila tissues and cell lines. PMID- 11396011 TI - Phosphatidylserine exposure and phagocytosis of apoptotic cells. PMID- 11396012 TI - The (Holey) study of mitochondria in apoptosis. PMID- 11396013 TI - In situ detection of dying cells in normal and pathological tissues. PMID- 11396014 TI - Model cell lines for the study of apoptosis in vitro. PMID- 11396015 TI - Programmed cell death assays for plants. PMID- 11396016 TI - Studies of apoptosis proteins in yeast. PMID- 11396017 TI - Methods to study cell death in Dictyostelium discoideum. PMID- 11396018 TI - Flow cytometric analysis of cell shrinkage and monovalent ions during apoptosis. PMID- 11396019 TI - Methods of study of tumor necrosis factor-related ligands in apoptosis. PMID- 11396021 TI - Feet first. PMID- 11396020 TI - Use of flow and laser-scanning cytometry in analysis of cell death. PMID- 11396022 TI - Are people ready for the job? PMID- 11396023 TI - Assessing the office. PMID- 11396024 TI - Oral fluid testing arrives. PMID- 11396025 TI - Rewriting the testing rulebook. PMID- 11396026 TI - SUBPART F--drug testing laboratories. PMID- 11396027 TI - Fostering first aid. PMID- 11396028 TI - Spill safeguards. PMID- 11396029 TI - Five steps to reduce cardiac arrest in your workplace. PMID- 11396030 TI - Being in the right place at the right time. PMID- 11396031 TI - The MERT team in action. PMID- 11396032 TI - The challenge of seating selection. PMID- 11396033 TI - The mat advantage. PMID- 11396034 TI - Bayer's comfort factor. PMID- 11396035 TI - Carpal tunnel syndrome recovery. PMID- 11396036 TI - Building safer operations. PMID- 11396037 TI - Constructing peace of mind. PMID- 11396038 TI - A cluster randomized controlled trial testing the effectiveness of a school-based dental health education program for adolescents. AB - OBJECTIVES: This trial investigated the value of a school-based dental health education program in terms of changes in knowledge, reported behavior, and plaque scores. METHODS: A total of 2,678 pupils with a mean age of 12.1 years attending 28 schools participated in a school-based dental health education program. The study used a cluster randomized controlled study design. The health service administrators stipulated that all participants receive the intervention; to meet this requirement, a rolling program of two six-month periods was utilized. During the first six months, half the adolescents received the intervention program, the other half acting as controls. Throughout a further six-month period, all participants received the intervention program. This research design allowed comparisons between participants receiving the program for six and 12 months. At baseline, six, and 12 months, a random subsample of 40 children in each participating school had their plaque scores recorded and a questionnaire was used to record their knowledge of dental health and reported dental behavior. RESULTS: The analysis used the subjects clustered within the schools, which were the units of randomization. The intervention program produced statistically significant improvements (P < .001) in knowledge about periodontal disease and the frequency of sugar intake and dental caries in both assessment time periods. The reported frequency of brushing did not change, but the group who had received 12 months of the intervention were more likely (P < .05) to brush for over a minute. At six months the early intervention group had a statistically significant, 13 percent reduction in the mean proportion of sites with plaque compared with the late intervention group (P = .043). This difference was sustained at 12 months (P = .037). CONCLUSION: This cluster randomized control trial demonstrated that the intervention program resulted in an improvement in knowledge of dental disease and an increase in the reported duration of brushing. These improvements were accompanied by a significant improvement in oral hygiene and a reported reduction in gingival bleeding. PMID- 11396039 TI - A pilot study of dental students' esthetic perceptions of computer-generated mild dental fluorosis compared to other conditions. AB - OBJECTIVES: Few studies have considered people's opinions about the esthetics of dental fluorosis. Assessments of fluorosis esthetics can be confounded by differences in a number of clinical factors, including tooth shape, color, contour, and gingival status. This pilot study compared esthetic perceptions of mild fluorosis and other conditions using computer-generated images made from a base set of normal appearing teeth. METHODS: Entering dental students (n = 61) completed questionnaires about four sets of paired photographs. Three sets consisted of fluorotic teeth (very mild to mild) versus other conditions (diastema, isolated enamel opacity, "normal"/control) and the other pair compared two presentations of mild fluorosis (generalized versus limited to incisal one third). Six questions, both qualitative and quantitative, were asked about each pair of photographs. RESULTS: Mild fluorosis was assessed less favorably than normal/control, midline diastema was less favorable than mild fluorosis, and mild fluorosis was less favorable than isolated opacity. CONCLUSIONS: This approach allows fluorosis to be better compared with other oral conditions because the images are standardized. Additional research with this method is warranted, including more variations in conditions, more comparisons, and other study populations. PMID- 11396040 TI - Managed care in dental markets: is the experience of medicine relevant? AB - OBJECTIVE: This paper reports on factors that predict the market penetration and growth into the market of both medical and dental managed care, and the relationship between the two. METHODS: Using data from the National Association of Dental Plans, the Interstudy Competitive Edge HMO Census, and the Area Resource File from 1987-95, we created an analytic data base covering the dental HMO market, the medical HMO market, dentist and physician supply, and regional market characteristics. Simple correlation analysis and multivariate linear regression using ordinary least-squares techniques were used to predict medical HMO penetration and dental HMO penetration in each state during 1994 and 1995. RESULTS: The results show that although the penetration of dental HMOs has been modest when compared to medicine, its growth is predictable by the same factors, and closely follows the pattern found in medical markets. CONCLUSIONS: Despite the observed relationship between medical and dental HMO penetration rates, there are potential barriers to managed care in the case of dentistry that may explain the slower growth to date, and that may ultimately decide the extent of managed care penetration into the dental market. PMID- 11396041 TI - Water consumption in the United States in 1994-96 and implications for water fluoridation policy. AB - OBJECTIVES: The purpose of this project was to describe current water consumption patterns and to compare them to findings from earlier studies. Current water consumption data also were used to reevaluate the association between water consumption and climate. These findings are of importance in estimating fluoride intake from fluoridated water. METHODS: Findings from the 1994-96 Continuing Survey of Food Intakes by Individuals (CSFII) were compared to those from two earlier dietary studies, the 1977-78 Nationwide Food Consumption Survey (NFCS) and the pioneering research of Galagan and colleagues in the 1950s. Food consumption data were analyzed for 14,619 persons with food and beverage intake data for two 24-hour periods in the CSFII. RESULTS: Increased consumption of infant formulas and decreased consumption of tap water and cow's milk were seen in the CSFII for infants compared to the NFCS. Older children and adults showed increased consumption of carbonated beverages and juices. While Galagan and colleagues found about a 60 percent increase in water consumption between the coldest (55 degrees F) and warmest (85 degrees F) conditions, only a 20 percent difference was seen between the winter and summer months in certain regions in the CSFII. CONCLUSIONS: No obvious strong or consistent association between water intake and month or season was apparent in these recent data. These findings are preliminary, and suggest that water fluoridation policy requires further research regarding water consumption and climate. PMID- 11396042 TI - Dental expenditures and source of payment by race/ethnicity and other sociodemographic characteristics. AB - OBJECTIVE: This study presents race/ethnic-specific distributions of dental expenditures and their sources of payment by socioeconomic characteristics among US working-age adults. METHODS: Data for persons aged 19-64 years from the 1987 National Medical Expenditure Survey (NMES) (n = 18,696) were used to calculate mean dental expenditures and their 95 percent confidence intervals. RESULTS: Dental expenditures were reported by 44.5 percent of participants. Non-Hispanic whites and persons with higher income were more likely to report dental expenditures than their counterparts. Among persons reporting expenditures, those with lower income had lower expenditures than higher-income persons. No differences in the amount of expenditures by race/ethnicity, sex, or employment status were observed. In all race/ethnic groups almost half the expenditures were paid out-of-pocket and one-third by dental insurance. CONCLUSION: While sociodemographic characteristics determined who had dental expenditures, they did not determine the amount or source of those expenditures. PMID- 11396043 TI - Hepatitis B prevalence and infection control among dental health care workers in a community in South Korea. AB - OBJECTIVES: The objectives of this study were to investigate the prevalence of hepatitis B vaccine and use of infection control procedures by dental health care workers in Taegu, South Korea. METHODS: Information was obtained with a mailed questionnaire sent to 300 private dental practices. Surveys were received from 177 dentists, 104 dental hygienists, and 46 dental assistants. All dental health care workers were asked to donate a blood sample for analysis of hepatitis B surface antigen (HBsAg) and antibody (anti-HBs). RESULTS: Vaccination against hepatitis B virus was reported by 63 percent of the respondents. About 40 percent of all participants answered that they were anti-HBs positive, while another 43 percent did not know their status. About 89 percent of dentists reported that they wore a mask with all patients, while hygienists (13.6%) and assistants (13.0%) were less likely to do so. Reported use of gloves (4.5%) and protective eyewear (14%) with all patients was extremely low in every group. Among the 56 dentists who were tested for HBV markers, 23 did not receive the vaccine against hepatitis B, and 13 percent (3 of 23) were positive for HBsAg as carriers. CONCLUSIONS: About 37 percent (120 of 327) of dental health care workers surveyed in this study in Korea did not receive the vaccine against HBV infection. Basic barrier techniques to prevent cross-contamination were not being used consistently. Nationwide guidelines for barrier techniques and hepatitis vaccinations should be developed and disseminated to dental personnel. PMID- 11396044 TI - Willingness to pay for periodontal therapy: development and testing of an instrument. AB - OBJECTIVES: The objective of this study was to develop and test the feasibility and validity of a willingness to pay (WTP) tool in a dental setting. METHODS: A questionnaire measured individuals' preferences among alternative treatments for periodontal disease and the maximum they would be willing to pay for their treatment of choice in terms of dental insurance premiums. The questionnaire provides detailed information, in probabilistic terms, of the risks and benefits of treatment choices for moderate to advanced adult periodontitis. It was pilot tested on 23 periodontal patients and 18 dental school faculty and staff. RESULTS: The majority (92.6%) felt the questionnaire was an accurate representation of treatments and outcomes, establishing face and content validity. In terms of construct validity, four hypotheses were tested: (1) manipulation of the outcomes of the preferred treatment led to a predictable shift in preferences for 38 subjects (92.7%); (2) although periodontal patients were not more likely to choose periodontal surgery than nonpatients (P = .14), those with a history of surgery were more likely to choose surgery again (P = .06); (3) WTP was positively related to income level (P = .05); and (4) subjects were willing to pay more for coverage for themselves than for others. Periodontal surgery was the preferred treatment for moderate to advanced periodontal disease, and was more strongly preferred than other choices (i.e., a higher WTP) for all income groups. The intraclass correlation coefficient for treatment preferences was 0.95 (P < .001) and the kappa for WTP was 0.78 (P < .001). CONCLUSION: This pilot study supports some of the criteria concerning validity of the WTP questionnaire to measure preferences for alternative periodontal therapies. Further testing on larger samples is required to confirm these results. PMID- 11396045 TI - Complex sampling: implications for data analysis. AB - Investigators in dental public health often use strategies other than simple random sampling to identify potential subjects; however, their statistical analyses do not always take into account the complex sampling mechanism. Often it is not clear whether a given strategy requires adjustment for stratification and/or cluster sampling of observations. We propose that the need for such adjustment depends on the primary study objective. As a general rule, we recommend that if the study goal is to estimate the magnitude of either a population value of interest (e.g., prevalence), or an established exposure outcome association, adjustment of variances to reflect complex sampling is essential because obtaining appropriate variance estimates is a priority. However, if the study goal is to establish the presence of an association, especially in a preliminary investigation of novel conditions or understudied populations, obtaining appropriate variance estimates may not be of primary importance; hence, adjustment of variances for complex sampling is not always required, but often is recommended. This paper describes several types of complex sampling designs, methods of adjusting for complex sampling strategies, examples illustrating the effect of adjustment, and alternative approaches for analysis of complex samples. PMID- 11396046 TI - Caries experience of urban and rural children in Saudi Arabia. AB - OBJECTIVE: Caries experience and restorative treatment needs often are reported to be higher in urban than rural areas in developing countries. The purpose of this study was to compare caries experience in 12- and 13-year-old children in urban and rural areas of Saudi Arabia, a young, oil-rich, developing country. METHODS: A random sample of 1,873 schoolchildren aged 12-13 years from urban and adjoining rural areas of 10 administrative regions in the Kingdom of Saudi Arabia were examined for dental caries. Questionnaires also were administered to elicit information on the frequency of snacking as well as consumption of sweets and soft drinks. RESULTS: No statistically significant differences were found between urban and rural children in caries experience in permanent teeth (mean DMFT = 2.69 in urban areas; 2.65 in rural areas), frequency of snacking, or frequency of consumption of sweets and soft drinks. The percentage of DMFT found to be D was equally high (> 89%) in both urban and rural areas, indicating a high level of restorative treatment need. CONCLUSION: In Saudi Arabia, an economically prosperous developing country, exposure to cariogenic diet in urban and rural areas does not differ. Also, a difference in caries experience in urban and rural areas often reported for developing countries does not apply to Saudi Arabia. PMID- 11396047 TI - Clinical manifestations and incidence of oculo-respiratory syndrome following influenza vaccination--Quebec, 2000. PMID- 11396048 TI - Progress towards Canadian target coverage rates for influenza and pneumococcal immunizations. PMID- 11396049 TI - The problem of the healing of intraaortic arterial prostheses in the treatment of abdominal aortic aneurysms. PMID- 11396050 TI - Posterior transverse plication: a technique for treatment of redundant internal carotid artery during endarterectomy. PMID- 11396051 TI - Diagnosis and treatment of Barrett's oesophagus. A general survey. AB - Barrett's oesophagus is an acquired condition with columnar metaplasia of the distal oesophagus. This condition represents the most serious consequence of chronic gastro-oesophageal reflux as it is associated with an increased incidence of oesophageal adenocarcinoma. Since the exact pathophysiology is not known, prevention is not possible. The diagnosis of Barrett's oesophagus requires the presence of intestinal metaplasia in at least one biopsy specimen from the lower oesophagus. Barrett's oesophagus is considered a premalignant condition and some cases progress from dysplasia to invasive adenocarcinoma. Medical or surgical antireflux treatment controls symptoms and oesophagitis, but Barrett's oesophagus remains. Patients are usually followed up by endoscopy for detection of dysplasia or early cancer. Several reports in the literature have assessed the effects of H2-blocker and proton pump inhibitors treatment on Barrett's epithelium, but none has clearly documented a significant and consistent regression of the metaplastic epithelium. Even with high doses of proton pump inhibitors given for a prolonged period of time, it does not appear that a significant regression of Barrett's epithelium can be achieved. Various studies have assessed the effects of antireflux surgery on the regression of columnar epithelium and dysplasia and its potential protective effect on the subsequent development of carcinoma. Overall, it appears from these reports that antireflux surgery, despite adequate symptomatic results, does not significantly and consistently lead to a reduction in length or disappearance of the Barrett's mucosa, and does not prevent the development of dysplasia and its progression to carcinoma. Recently, numerous reports have documented the regression of Barrett's mucosa by using various experimental techniques: these thermal therapies focus on the removal of the columnar epithelium with restoration of the squamous epithelium. Technological advances including laser and especially photodynamic therapy have allowed for endoscopic mucosal ablation. Long-term results are more encouraging when this mucosal ablation is associated with antireflux medical or surgical therapy. Currently, none of these approaches can obviate the need for continued endoscopic surveillance; however the photodynamic therapy seems to be a promising alternative in the future. PMID- 11396052 TI - Abdominal compartment syndrome. AB - Abdominal compartment syndrome may be defined as the deleterious pathophysiologic consequences of a significant increase in intra-abdominal pressure. These alterations can affect respiratory mechanics, cardiovascular system, regional blood flow, renal function, urine output, and intracranial pressure. Although the syndrome may be associated with many clinical situations, the most common are severe abdominal trauma and ruptured abdominal aortic aneurysm. Diagnosis depends upon recognition of the clinical syndrome followed by an objective measurement of intra-abdominal pressure, the most common being the measurement of bladder pressure. Treatment consists of adequate fluid resuscitation and surgical decompression when necessary. PMID- 11396053 TI - Prophylactic appendectomy: is it worth to be done? AB - There are still many controversial issues in prophylactic appendectomy. In this retrospective study, we have appraised the reasons and results of prophylactic appendectomies performed between January 1997 and August 1999. Sixty four prophylactic appendectomy cases were included in this study and all resected specimens were submitted to histopathological analysis. In 23 patients with incisional hernias, a prophylactic appendectomy was performed as, in the future, dense adhesions may lead to difficult appendectomy. Additional appendectomy had no negative effect on perioperative complications, compared with the patients without appendectomies. In conclusion, in the presence of optimal conditions, prophylactic appendectomy can be performed safely. PMID- 11396054 TI - What did the laparoscopic Nissen approach of the gastro-oesophageal reflux really change for the patients 8 years later? AB - BACKGROUND: Nissen fundoplication (NF) is recognized as the surgical treatment of the gastro-oesophageal reflux disease (GERD). NF can be achieved either by open surgery or by laparoscopic approach. METHODS: From 1987 to 1997, 210 patients were treated for GERD by NF: 61 by open and 149 by laparoscopic approach. All the patients were followed more than 1 year and were scored by clinical assessment (Visick scale adaptation). In case of Visick score > 1, GI-endoscopy, X-ray series or 24-hour pH-study complete the evaluation. RESULTS: The operative time was comparable between both groups. The postoperative recovery was statistically faster in the laparoscopic group (p = 0.0001). The mean time of follow-up was 6 years after open NF and 4 years after laparoscopic NF. After open NF or laparoscopic NF, 72% and 67% of the patients are respectively scored Visick 1, 13% and 21%--Visick 2, 6.8% and 6%--Visick 3 and 8.2% and 6%--Visick 4 (NS). Patients with recurrence of GERD were scored Visick 4, so failure of the surgical treatment is observed in 5 patients after open NF and 9 patients after laparoscopic NF. The occurrence of incisional hernia was significantly higher in the open group (p = 0.0001). CONCLUSION: NF remains a safe procedure for surgical treatment of GERD and can be achieved by laparoscopic approach with comparable results to those by open laparotomy. In our experience, the advantages of the laparoscopic approach is a faster postoperative recovery and a lower risk of incisional hernia. PMID- 11396055 TI - Hyperbaric oxygen (HBO) as useful, adjunctive therapeutic modality in compartment syndrome. AB - The authors describe a compartment syndrome progressively developed after a long term surgical procedure, with a patient positioned in supine position with calf rest, who was successfully treated with hyperbaric oxygenation. This approach saved the patient from a more invasive therapeutic intervention. PMID- 11396056 TI - Fusiform aneurysm of the internal jugular vein: an unusual cause of neck swelling. AB - Aneurysmal anomalies of the internal jugular vein are very uncommon. We report two cases of internal jugular vein aneurysm and discuss the clinical pattern, pathology, aetiology and management. PMID- 11396057 TI - Primary angioleiomyosarcoma of the lesser omentum. AB - Primary omental leiomyosarcomas are rare intra-abdominal tumours. A few cases only have been reported in the literature. We present a case of a 79-year old woman, whose tumour was surgically removed. We include a hort review of the literature. PMID- 11396058 TI - Gastric incarceration and perforation following posttraumatic diaphragmatic hernia. AB - We report the case of a 36-year-old male patient who developed gastric incarceration and perforation in a diaphragmatic hernia 8 months after an automobile accident. During emergency surgery, protrusion of the stomach into the thoracic cavity and perforation on the anterior aspect of the stomach were noted. The gastric perforation and the diaphragmatic defect were closed. During the postoperative course, the patient developed sepsis and coagulopathy that subsided following medical therapy. In order to prevent severe complications, surgery is indicated as soon as conclusive diagnosis is made. PMID- 11396059 TI - Spinal cord stimulation in critical limb ischemia. PMID- 11396060 TI - [Emergency surgery: the extraordinary trip of an idea]. PMID- 11396061 TI - Long-term results of aggressive surgical treatment of primary and recurrent retroperitoneal liposarcomas. AB - The aim of this study was to establish the role of surgery in the treatment of retroperitoneal liposarcomas. Data concerning 28 patients submitted to surgery for retroperitoneal liposarcoma in our department over the period from 1972 to 1999 were reviewed retrospectively and analysed. Seventy-four operations were performed; in 54% of the operations it was necessary to resect contiguous organs (kidney 60%, colon 50%, adrenal gland 35%). In 89%, grossly curative resection was achieved at the first operation; 20 patients had at least one local recurrence after first operation (median time interval: 22 months). The mean follow-up was 80 months; median survival time was 51 months and 5-year actuarial survival time 51%. Patients with low-grade liposarcoma showed a statistically significant improvement (P < 0.001) in median survival (153 months) versus those with medium- (37 months) and high-grade sarcomas (8 months). At present surgery is still the treatment of choice in the treatment of primary and recurrent liposarcoma; in the case of low-grade liposarcomas especially, an aggressive surgical approach can result in long-term survival. PMID- 11396062 TI - Early surgical treatment of acute cholecystitis. A retrospective comparative study of the laparoscopic and open approaches. AB - The aim of this retrospective study was to compare the results of laparoscopic and open early cholecystectomy in patients with acute cholecystitis. From January 1997 to October 2000, 168 patients underwent cholecystectomy in our institution. Of the 35 patients (20.8%) with acute cholecystitis, 20 patients (57.1%) were operated on laparoscopically and the other 15 patients (42.9%) with the traditional open approach. The two groups were similar in terms of age, sex and onset of symptoms. The postoperative morbidity was 15.0% in the laparoscopic group versus 40.0% in the open group. The average postoperative hospital stay in the laparoscopic group was 5.1 days as compared to 10.5 days in the open group (P = 0.013). The conversion rate to laparotomy was 5.0% (1 case). At follow-up there has been one case of incisional hernia in the open group. Early laparoscopic cholecystectomy for acute cholecystitis was associated with a lower postoperative morbidity rate and significantly earlier patient discharge. PMID- 11396063 TI - [Experience of 10 years with the surgical treatment of lung cancer in elderly patients]. AB - The surgical treatment of lung cancer in elderly patients is correlated with a lot of complications that in most cases are cardiopulmonary type; for this reason, in past times these patients were left out of the surgical treatment. Today, the progress of surgical, diagnostic and anesthetic techniques permit to execute pulmonary resections in patients older than 70 years too. The aim of this study is to evaluate postoperative complications and long-term and long-term survival in patients under and over the age 70. Between January 1990 and June 2000 we have assessed 172 patients with lung neoplasm. We have divided patients in two groups: those younger than 70 years of age (group 1, n = 119) and those older than 70 years of age (group 2, n = 53). Postoperative mortality for group 2 was 7.5% and for group 5.1%. The overall postoperative complication rate for group 2 was 30.2% and for group 1 10.9%. Within group 1 main complications have been of surgical type (61.5%) while in group 2 medical type (cardiopulmonary), with higher frequency in patients ASA 3 or ASA 4. The mortality at 12 months for non-neoplastic causes was 10.2% for group 2 and 2.5% for group 1. On the contrary, the mortality at 12 months for neoplastic causes was 8.2% for group 2 and 6.7% for group 1. We haven't noticed an important correlation between the extension of the resection and the recurrence of disease. The survival at 3 years was 46.5% for group 1 and 41% for group 2 and associated with neoplastic causes. PMID- 11396064 TI - [Results of the surgical treatment of primary gastric lymphoma]. AB - The aim of the study was to verify the long term results obtained in primary gastric lymphoma with a strategy consisting in surgery as first-line treatment. Over the period from January 1988 to December 1999, 44 patients with histologically proven primary gastric lymphoma underwent surgical treatment in the First Department of General Surgery of the University of Verona. Tumours were staged according to the Ann Arbor classification and divided, according to the Kiel classification, into high- and low-grade lymphoma. Patients received adjuvant chemotherapy depending on the grade of malignancy and/or completeness of resection. Of the 44 patients, 40 (90.9%) underwent curative resections, i.e. with complete macroscopic and microscopic tumour removal (R0), consisting in total gastrectomy in 34 cases and subtotal gastrectomy in 6. Twenty-five of 40 patients had stage IE and 15 stage IIE tumours. Adjuvant chemotherapy was given to 33 patients (30 high-grade lymphomas and 3 low-grade lymphomas with N2 metastases). The overall cumulative 10-year survival rate in patients who underwent R0 resection was 79% without any significant differences in 10-year survival between patients with high- and low-grade malignancy (both 79%; P = 0.582) or between patients with or without lymph node metastases (91% and 70%, respectively; P = 0.426). In conclusion, the present investigation suggests that surgery yields prolonged complete remission in a high percentage of patients affected by gastric lymphoma irrespective of histopathologic grade of the disease and nodal involvement. PMID- 11396065 TI - [Non-surgical treatment of liver trauma]. AB - The aim of the study was to assess the role of non-operative treatment in haemodynamically stable patients with liver trauma. Over the period from 1996 to July 2000, out of a total of 2,048 patients with abdominal trauma, 124 open and 1,924 closed, we observed 77 hepatic lesions, consisting of 55 closed traumas and 22 penetrating traumas. Non-operative treatment was implemented in 18 patients (32.7%) with closed liver traumas. In addition to serial clinical examinations of the abdomen, the patients receiving non-operative treatment were submitted to thorough haemodynamic monitoring and complete blood counts in the intensive care unit. After an abdominal CT scan at entry, patients were submitted to abdominal ultrasonography 6, 12 and 24 hours after admission. Only two patients required transfusions, one presenting a pelvic fracture and the other a triple fracture of the femur, tibia and fibula. There was no mortality. A biloma was present in one case, successfully treated by means of a US-guided drainage puncture. It is patients with major cranial traumas that pose most problems for conservative treatment. Fifty percent of non-therapeutic laparotomies in our series were performed in patients with severe cranial traumas. It is precisely in these patients that an improvement in diagnostic capability is most desirable. PMID- 11396066 TI - [Biliary lesions after laparoscopic cholecystectomy]. AB - Laparoscopic cholecystectomy is the suitable treatment for symptomatic cholelithiasis, even if the incidence of biliary lesions following this procedure may be up to threefold higher than that of open cholecystectomy. We report our experience concerning the incidence, aetiopathogenesis, diagnosis and treatment of complications in a homogeneous group of laparoscopic cholecystectomies. In a total of 492 laparoscopic cholecystectomies only three bile duct lesions were observed (0.6%); they were classified according to Bismuth and re-assessed according to Strasberg. They consisted in two biliary leakages and one bile duct stricture. All patients were evaluated by full blood test, ultrasonography and endoscopic retrograde cholangiopancreatography. Endoscopic treatment was successful in the two patients with biliary leakage, while the patient with a stricture required surgical therapy. In conclusion, we suggest that a correct knowledge of the aetiopathogenesis together with a multidisciplinary approach to the diagnosis appear to be the best method for the detection, complete classification and most suitable treatment of symptomatic cholelithiasis. PMID- 11396067 TI - [Laparoscopic fundoplication in gastroesophageal reflux disease: reflexions on a personal caseload]. AB - Ninety-two patients with severe, proton-pump-inhibitor-dependent gastro oesophageal reflux disease were submitted to surgery and operated on by the same surgeon (SC) over the past 7 years (mean age: 42; range: 23-74 years). Partial fundoplication was performed in 14 patients with impaired oesophageal motility, while 78 total fundoplications were done in the others, 51 without, and 27 with division of the short gastric vessels. The mean follow-up was 29.5 months (range: 1-85 months). Conversion to open surgery was necessary in 6 patients (all in the first 40 cases). Perforation of the gastric fundus and early migration of the stomach into the mediastinum were the two most important complications observed. The mortality was nil. 39% of the patients complained of postoperative dysphagia but only five required endoscopic (4) or surgical (1) treatment. The percentages of dysphagia after partial fundoplication and total fundoplication with or without division of the short gastric vessels were 28%, 37% and 47%, respectively. In 83.7% the patients were satisfied with the clinical results and in 84% of cases medical treatment was avoided after surgery. On the basis of these data, laparoscopic surgery appears to be a good option for gastro oeophageal reflux disease in selected patients with a poor response to, or dependent on medical treatment. However, the results of surgery may be subject to the limitations of a learning curve, as in all complex laparoscopic procedures. PMID- 11396068 TI - [Indications for preventive tracheostomy in the surgical treatment of non neoplastic thyroid disease]. AB - In thyroid surgery, an emergency tracheostomy is mandatory, as is well known, in acute pre- and postoperative asphyxia due to obstruction of the upper airways, especially in cases of malignant tumours. In particular conditions, in the presence of benign thyroid disease and when there is no reduction in the laryngo tracheal lumen, tracheostomy has an important precautionary function following total thyroidectomy. The authors describe their experience with 6 patients treated by precautionary tracheostomy after total thyroidectomy for benign thyroid disease, analysing the indications and the advantages of this method. PMID- 11396069 TI - [Effects of surgical repair of first degree varicocele on gonadal function and testicular atrophy]. AB - Varicocele is one of the main causes of male infertility. This pathology, in fact, is responsible for progressive anatomical and functional testicular damage. Constant monitoring of subjects suffering from varicocele is therefore necessary. In the present study, we evaluated the effect of varicocele surgical treatment on seminal parameters, testicular growth and pregnancy rate in patients with grade I varicocele. We operated on 47 patients with left grade I varicocele associated with sperm abnormalities and with testicular hypotrophy. A high ligation of the internal spermatic vein and all its collaterals was performed in all patients. The postoperative follow-up showed a significant improvement in seminal parameters (motility, morphology and sperm count) and in testicular volumes. Moreover, a pregnancy rate of 58% was observed. These results confirm the observations of other investigators and suggest that surgical therapy of varicocele is capable of improving seminal parameters, of reversing varicocele related testicular hypotrophy and of achieving high pregnancy rates even in patients with grade I varicocele. PMID- 11396070 TI - [Ambulatory surgery: preoperative assessment]. AB - Over the past 20 years public health expenditure has progressively increased for various reasons. With the aim of reducing public health expenditure and hospital stays, alternative models of public health care have been more widely adopted over the past two decades: Day Hospital, day surgery and ambulatory surgery. What is meant by ambulatory surgery is the clinical, organisational and administrative possibility of performing surgical operations and/or invasive and semi-invasive diagnostic procedures without hospitalisation of patients, in doctors' surgeries, outpatient departments or protected outpatient facilities. For both the patients and the centres providing the service to be able to exploit all the advantages stemming from this kind of health care provision, careful patient selection is mandatory, including assessment of the impact of any potential concomitant pathologies. Starting from their own personal experience and existing reports in the international literature, the authors analyse the impact that concomitant pathologies may have on patient selection for ambulatory surgery. The authors conclude that thorough preoperative evaluation of all the possible variables involved is the only way of ensuring the success of ambulatory surgery. PMID- 11396071 TI - [Proctologic day-surgery. Results of 2000 surgical interventions]. AB - The aim of the study was the evaluate of results of 2000 surgical operations for ano-rectal disease performed in the day-surgery setting (7-24 hours hospital stay) with improvement of both cost effectiveness and patient comfort. From January 1980 to December 1998, 2000 patients underwent surgical operations: 1011 for haemorrhoids; 708 for anal fissure; 172 for fistula in ano; 80 for pylonidal disease; and 45 for anal stenosis. 97.6% of patients were operated on with loco regional anaesthesia; the others with narcosis and peripheral anaesthesia. The hospital-stay was 24 hours in 697 patients (34.5%), while 1319 (65.5%) operated on under loco-regional anaesthesia were hospitalised for 7-10 hours. Three patients (0.2%) developed acute hemorrhage after hemorroidectomy during the immediate postoperative period. They underwent reintervention under general anaesthesia with a hospital stay of 7 days. Four patients (0.6%) with perianal abscess after internal sphincterotomy underwent incision 10 days after the operation. Two patients with perianal hematoma after sphincterotomy prolonged the hospital stay for three days. In 1048 patients (51.9%) clinical recovery was observed at first follow-up (7 days); 48% had recovered at the 2nd follow-up (14 days). In 1608 patients (98%) anatomical recovery was observed at the follow-up three months after surgery. Patient satisfaction 6 month after operation was high in 79%; good in 27%; low in 1%. These results seems confirm the feasibility of proctological day surgery in almost all patients, with both a considerably cost reduction and enhanced patient comfort and compliance. PMID- 11396072 TI - ["Difficult" venous ulcers of the legs]. AB - The aim of the study was to analyze the diagnostic and surgical criteria in cases of "critical" venous leg ulcers. This report describes the management of 200 patients with trophic lesions of the lower limbs. Patients with venous leg ulcers were identified with the aid of ultrasonography and echocolor Doppler. Among these patients, according to the chronic evolution of the lesions, we identified all those with critical venous leg ulcers. The management of the lesions involved both medical and fairly aggressive surgical treatment with the use of general anaesthesia and deep surgical toilette of the ulcer. In the cases with poor epithelialisation we adopted oxygen therapy and skin grafts using the "mesh graft" technique. Critical venous leg ulcers were identified in 23 patients (19%). After surgical treatment, we observed complete epithelialisation of the lesion only in 5 cases (21.7%). In 13 patients (56.6%) the use of oxygen therapy was necessary and epithelialisation occurred on postoperative day 12. Skin grafts were used in 5 cases (21.7%). Only one recurrence was observed one year after operation in an area adjacent to the site of the graft. The "golden goal" of critical venous leg ulcer management is to establish a correct aetiological diagnosis, distinguishing between venous leg ulcers and other trophic lesions of the lower limbs. The medical and surgical treatment described in this report reduce healing time and the social costs of the disease, without any significant complications for the patient. PMID- 11396073 TI - [Ultrasonography of normal vermiform appendix]. AB - The aim of this study was the detection and visualisation of the normal vermiform appendix and its characteristics by ultrasonography in adults with no clinical suggestion of acute or chronic abdominal disease. A prospective study was performed in 200 subjects. The graded-compression ultrasonography technique was used to explore the lower right quadrant of the abdomen and the pelvis. The examination was performed using a 4 MHz sector array and 7.5 MHz linear array transducer. In a few cases, a 10 MHz linear array transducer was used. The appendix was visualized in 54% of patients. In all cases where the appendix was visualized it was found to be either on the ileo-psoas muscle or directly beneath the abdominal wall. The ileo-caecal valve was visualized in 78% of cases. The transverse diameter was found to be no greater than 6.5 mm except in three cases that had a diameters ranging from 7 to 9 mm. Diameter variability along the length of the same appendix was demonstrated in 5% of subjects. Wall thickness was no greater than 2.5 mm. Our experience suggests that graded-compression ultrasonography is a valuable procedure for detecting the vermiform appendix more frequently than has been previously reported. The patients physical constitution and the anatomical location of the vermiform appendix were found to be important factors affecting the ability to visualize the vermiform appendix. The ability to visualise the normal vermiform appendix ultrasonographically supports the clinical diagnosis and excludes acute appendicitis. PMID- 11396074 TI - [Bilateral pheochromocytoma associated with duodeno-jejunal GIST in patient with von Recklinghausen disease: report of a clinical case]. AB - The authors present the case of a 60-year-old male patient suffering from von Recklinghausen's disease (neurofibromatosis type I, NF1) with bilateral pheochromocytoma and occasional intraoperative reports of duodenojejunal GIST (GastroIntestinal Stromal Tumour). Through a review of the literature the authors analyze the frequency and the features of bilateral pheochromocytoma and its rare histological variant, the so-called composite pheochromocytoma, characterized by the combination of pheochromocytoma and ganglioneuroma or ganglioneuro-blastoma. Bilaterality of pheochromocytoma is more frequent in patients with familiarity for pheochromocytoma without NF1. Composite pheochromocytoma accounts for about 3% of total pheochromocytomas. In addition, the authors summarize the present knowledge about gastrointestinal stromal tumours and investigate the possible association between them and NF1 or pheochromocytoma, concluding that any such association is purely casual, while confirming the well known, genetically determined association between NF1 and pheochromocytoma. PMID- 11396075 TI - [Pancreatic cystadenoma: report of 2 clinical cases and review of the literature]. AB - The Authors present two rare cases of pancreatic serous cystadenomas. Preoperative diagnosis of these neoplasms is very difficult despite the routine use of echotomography and CT, because the definitive diagnosis is provided only by histological examination of the surgical specimen. General agreement exists as to surgical indication. Surgical tactics depend on tumour localization: duodenocephalopancreatectomy for tumours of the head of the pancreas, and distal pancreatectomy for tumours located in the tail, as can be seen from a review of the recent literature. In both cases reported, the neoplasm was located in the pancreatic body. The treatment consisted in distal pancreatectomy. Splenectomy was necessary only in one of the two cases. During the follow-up the first patient died after 11 years without relapse. The second died 10 months postoperatively of an undifferentiated low rectal cancer. CT scans and sequential laparotomy showed no local relapse of the pancreatic tumours. PMID- 11396076 TI - [Perforated duodenal diverticulum: report of a case]. AB - After the colon, the duodenum is the most common site of diverticula. Duodenal diverticula can be divided into two types: intraluminal or extraluminal. The latter are more frequent, with a prevalence ranging from 0.6 to 27% in relation to the diagnostic methods utilized. Females are more often affected than males. About 70-75% of extraluminal duodenal diverticula are located in a circular area centred around the ampulla of Vater within a radius of 2-3 cm; these are defined as periampullary or juxtapapillary. Perforation is the rarest type of complication and can simulate different clinical conditions. CT plays a fundamental role in diagnosis also in relation to the different diverticular topography. Perforation is an indication for emergency surgery. The authors describe the clinical case of a duodenal diverticulum containing the outlet of the papilla, complicated by perforation; CT showed retroduodenal fluid and free air. Emergency surgery with an external biliary drainage, naso-biliary probe, and a diverticulo-jejunostomy on a Roux-en-Y defunctionalised loop, resolved the condition. PMID- 11396078 TI - [Informed consent in the light of the new medical deontologic code and of the state's laws]. AB - The Authors in the light of the legislative directives as well as of some juridical judgements and the new deontological code for medicine, especially from the articles 30 to 35, consider how information should be given to the patients. Furthermore the Authors underline how a correct information does not exclude the explicit consensus expressed by the patient before being submitted to any diagnostic and/or therapeutic treatment and how the patient's consensus should be collected. In addition the Authors briefly examine some juridical judgements that in the last years have been cause of tensions for physician and emphasize some of the fundamental rights of citizen in an up to date view of the medicine. PMID- 11396077 TI - [Endometriosis of umbilical cicatrix: a clinical case]. AB - The Authors report a case of umbilical endometriosis in a 46-year-old patient. The woman came in for observation describing acute pain in the vicinity of the umbilical scar. The pain was occasional at first and then became steady and increased at the time of menstruation. Medical examination revealed a left paraumbilical nodule, measuring 1 cm in diameter. The patient underwent surgical treatment: the nodule was excised and the subsequent histological examination was diagnostic for umbilical endometriosis. The surgical excision was effective: at follow-up 3 months later, there was no recurrence and the patient was in good general condition. PMID- 11396079 TI - [Multicenter prospective study of informed consent in general surgery]. AB - To understand the level of acceptance, awareness and usefulness of informed consent, a group of 119 patients (59 men and 60 women) from different types of hospitals were given a questionnaire which required only 'YES or NO' answers, both before and after surgery. The questionnaire concerned the patient's knowledge about pathology, operative risks, approval, anxiety caused, understanding of information received and consent given, and also if he would inform a relative in the same condition. From the analysis of the results it was established that: the more information a patient has about his illness and operation risks, the more he will want to have; the less he knows the less he will want to know, and he will also have more faith in the doctors. Some patients would not inform a relative with a similar pathology. To conclude, informed consent, instead of being a right of the patient is progressively becoming more a right of the doctor. It does not have any real effect on the patient's choice but is useful, as it represents a moment of personalised attention from medical personnel, though the patient may not completely understand the information received. There are few advantages in strictly medical terms but informed consent has increased malpractice litigation. PMID- 11396080 TI - Point-of-care testing. An overview and a look to the future. AB - Point-of-care testing is an emerging specialty in laboratory medicine and has attracted considerable interest in the medical literature. This article presents a general overview of point-of-care testing including its history, menu, clinical utility, aspects of analytical performance, regulatory issues, and management in the hospital setting. Some perspectives concerning the possible future of point of-care testing are also presented in view of current issues in the hospital and clinical laboratory. PMID- 11396081 TI - Regulatory compliance for point-of-care testing. A perspective from the United States (Circa 2000). AB - Regulatory guidelines concerning point-of-care testing have had a major impact on the development and evolution of these technologies. Beginning with mandates under CLIA '88, hospital accreditation organizations have developed guidelines for the management of point-of-care testing to ensure quality testing. Over the past decade, most hospitals have learned to manage these new technologies as part of the laboratory quality assurance program. Issues concerning regulatory compliance continue to influence the development and use of point-of-care technologies providing opportunity for continuous quality improvement and improved patient care. PMID- 11396082 TI - Management of a point-of-care program. Organization, quality assurance, and data management. AB - Management of a POCT program can be difficult. Cooperation and communication are essential. Creating an interdisciplinary POCT committee is key to successful management. Each discipline through its expertise and common goal to provide quality patient care contributes to the success of the program. The development of a comprehensive QA program ensures that POCT results are accurate, reliable, and performed by competent personnel, and that regulatory compliance is achieved. Continuous quality improvement diminishes the POCT program's potential for undesirable outcomes. Ultimately, the patient benefits, with quality patient care and improved outcomes. Improved data management is necessary with the increased demands for point-of-care tests and the volume of data already generated to ensure quality of results and still meet regulatory guidelines. More automated systems are needed to eliminate labor- and time-intensive tasks. The function of downloading data requires complete automation and transparency for nursing and other testing personnel to gain complete cooperation. The future for data management, however, looks promising for meeting the needs of today. PMID- 11396083 TI - Point-of-care testing. Cost issues and impact on hospital operations. AB - Advancing technology has allowed for the migration of laboratory testing from the central laboratory to the near-patient setting, leading ideally to a shorter therapeutic turnaround time. This potential benefit, however, comes with a price tag. Assessing the cost-benefit ratio on a per test basis cannot effectively be done in a generalized manner, because each hospital must evaluate this with respect to its own unique circumstances. There are, however, certain outcomes, such as decreased LOS and decreased blood-product usage that, if achieved, far outweigh the cost of POCT, justifying its use. Any hospital attempting to implement POCT must also realize that hospital operations are affected by such use and that adjustments and careful laboratory oversight are required. PMID- 11396084 TI - Point-of-care testing. Impact on medical outcomes. AB - There is now clear evidence that POCT has a positive benefit on morbidity and mortality. In addition, there are other tangible benefits that may themselves influence morbidity and mortality, e.g., reduced blood sample requirement in pediatrics, reduced length of stay, and greater doctor and patient satisfaction. These benefits accrue from the ability to make decisions and implement the appropriate intervention more quickly. It has also been demonstrated that POCT can facilitate improved patient motivation and satisfaction and thereby compliance with a prescribed disease management strategy. Improvement in health outcome, morbidity, and mortality can only be achieved, however, when the diagnostic and therapeutic interventions operate in concert. A review of the literature on medical outcomes of POCT has demonstrated the complexity of establishing evidence and the paucity of robust literature that exists at the present time. It is hoped, however, that the reader will appreciate how important it is to stress the role of a diagnostic test in decision making, to ensure that decisions are made and that benefits will be achieved. From a more pragmatic standpoint, however, it is hoped the reader will see the potential value of the arguments put forward in favor of implementing POCT when submitting a business case to a funding authority; the fact that the cost of POCT may be greater should not be a deterrent. PMID- 11396085 TI - Bedside glucose testing. Applications in the home and hospital. AB - Point-of-care testing is an increasingly popular means of delivering diagnostic testing closer to the site of patient care. Although point-of-care glucose testing devices have been around for over a decade, concerns about the quality of the results still plague the industry. Changes in federal and state laws that treat point-of-care testing as an extension of the core laboratory promise to improve the quality of testing. The most challenging aspect of quality testing is selection of the appropriate method (core laboratory versus point-of-care test) for the optimal patient outcome. PMID- 11396086 TI - Acute care testing. Blood gases and electrolytes at the point of care. AB - The standard turnaround time for acute care laboratory testing in tertiary care institutions is typically less than 15 minutes for blood gas or electrolyte values. From a clinical perspective, however, the desirable turnaround time is more on the order of 5 minutes, and this is technically achievable. The 15-minute standard can be met with strategically located STAT laboratories. To achieve a turnaround time of 5 minutes, it is necessary to move the "laboratory" closer to the patient and to have more than one instrument available. This latter configuration is called near or bedside patient testing. Why the 5-minute standard is not used universally throughout the nation is probably related to differing perspectives on "cost" and "quality." As manufacturers, hospitals and laboratories address the issue of rapid turnaround time in acute care settings, the 5-minute standard may become more widespread. Direct costs have been decreasing as more manufacturers enter the market for acute care testing. The overall quality is also improving, not only in the engineering features built into the instruments, but also as nonlaboratory staff gain skill in performing the testing. As more sites implement POCT, standards and guidelines for managing testing outside of the laboratory are being established. Solutions to preanalytic problems are being developed and implemented. POCT testing for blood gases and electrolytes was once considered to lie in the future but is now commonplace and may one day become the standard of care. PMID- 11396087 TI - Coagulation point-of-care testing. AB - With PT point-of-care devices, further study is needed to fully evaluate the safety and efficacy regarding home self-monitoring of oral anticoagulant therapy. Point-of-care PTT testing is also undergoing evaluation. In contrast, the ACT is commonly in use, despite its limitations, at least partly because of the lack of a readily available, inexpensive alternative with sufficient turnaround time. Several platelet function point-of-care devices have also become available, but their role in clinical care is not yet well defined. PMID- 11396088 TI - Testing for cardiac markers at the point of care. AB - Measurement of cardiac markers at the point of care is feasible, diagnostically accurate, and, when combined with appropriate decision-making algorithms, cost effective. It may be the only way to deliver results rapidly enough for therapeutic intervention. PMID- 11396089 TI - Toxicology and drugs of abuse testing at the point of care. AB - Point-of-care methods have been developed for the detection of abused drugs in urine. This article identifies substances, illicit and licit, that are targeted by testing for reasons of medical care, drugs policy, or law enforcement. The advantages and disadvantages of point-of-care testing, and clinical interpretation of test results, are discussed. Point-of-care breath alcohol analysis and its use as an alternative to laboratory blood measurement are also included. PMID- 11396090 TI - Provider-performed microscopy. AB - The category of provider-performed microscopy was defined in the Federal Register to provide a unique regulatory approach for bright-field and phase-contrast microscopy performed by a physician, dentist, or midlevel practitioner examining labile specimens. A CLIA certificate for this category of testing is available, which also permits the performance of waived tests. Because these provider performed microscopy procedures do not have quality-control materials available, special challenges are encountered in establishing and monitoring such testing within a hospital. Vigilance is required to maintain a provider-performed microscopy program within a hospital. PMID- 11396091 TI - Selected topics in point-of-care testing. Urinalysis, pregnancy testing, microbiology, fecal occult blood, and other tests. AB - The menu of tests available at the point of care continues to expand. Many of these tests present unique opportunities to deploy point-of-care technologies in the home, outpatient, and hospital settings. The future will inevitably create new applications for POCT. The success of these efforts will be in part determined by how technologies for specific applications are selected and by the ability to manage new point-of-care tests as part of an organizational point-of care program. The possibilities to improve patient care are obvious, provided adequate attention is paid to ensuring quality testing and to managing patient data from these novel technologies. PMID- 11396092 TI - The epidemiology of rheumatoid arthritis. AB - Studies of the descriptive epidemiology of RA indicate a population prevalence of 0.5% to 1% and a highly variable annual incidence (12-1200 per 100,000 population) depending on gender, race/ethnicity, and calendar year. Secular trends in RA incidence over time have been shown in several studies, supporting the hypothesis of a host-environment interaction. People with RA have a significantly increased risk of death compared with age- and sex-matched controls without RA from the same community. The determinants of this excess mortality remain unclear; however, reports suggest increased risk from gastrointestinal, respiratory, cardiovascular, infectious, and hematologic diseases among RA patients compared with controls. Despite extensive epidemiologic research, the etiology of RA is unknown. Several risk factors have been suggested as important in the development or progression of RA. These include genetics, infectious agents, oral contraceptives, smoking, and formal education. Epidemiologic research is an essential contributor to our understanding of RA. PMID- 11396093 TI - Comorbidity in rheumatoid arthritis. AB - It is increasingly clear that coexistent disease plays a pivotal role in RA outcome and that efforts aimed at specifically addressing these comorbidities need to be aggressively sought, investigated, and implemented once proven effective. RA-associated costs are currently increasing at twice the rate of the medical care index. Comorbidity in the setting of RA independently predicts disease-associated disability (a major cost component) and mortality, underscoring the need for a more comprehensive approach to RA, one that adequately addresses disease-specific comorbidities. At present, many primary and secondary preventative measures (Table 1) for RA-specific comorbidities remain largely unproved and require rigorous investigation in a randomized prospective fashion. Despite this ongoing need, advances are being made in our understanding of the underlying pathogenesis of these comorbid conditions and their relation with RA. This improved understanding should translate into further effective interventions. Bisphosphonates, for instance, have been shown to be effective in the prevention of GIOP and associated fractures. The past several years have seen other exciting therapeutic advances in RA. DMARD combinations have been shown to be more effective and no more toxic than MTX monotherapy. In addition to the recent release of COX-2 NSAIDs, three new disease-modifying agents (leflunomide, etanercept, and infliximab) have been added to the therapeutic armamentarium; these are options that have markedly changed the treatment landscape in RA. Although these important advances have generated much deserved optimism, the precise effect that these agents may have on RA-specific comorbidity remains to be seen. The next decade should prove to be an exciting time in RA management. Better identification, understanding, and management of RA comorbidities have great potential to improve quality of life and survival among our patients with RA. PMID- 11396094 TI - The role of the DR4 shared epitope in selection and commitment of autoreactive T cells in rheumatoid arthritis. AB - The mechanistic basis for HLA associations with RA is still unknown in spite of 20 years of disease association studies and a detailed characterization of HLA class II alleles associated with disease. Analysis of the structural interactions between DR4 susceptibility molecules and T cells specific for the peptide-MHC complex suggests a mechanism for directed T-cell selection and amplification in which RA-associated genetic polymorphisms bias intermolecular recognition. New immunologic models for illustrating the importance of regulated thresholds for T cell activation based on avidity between the TCR, MHC, and peptide offer insight into a potential mechanism in which the disease-associated HLA molecules create an autoimmune-prone individual by virtue of a biased TCR selection and T-cell amplification process. New tools such as the use of HLA-DR4 tetramers provide the ability to identify and monitor the presence of such autoreactive T cells in the periphery of individuals and patients and should assist in further testing of the multistep model for RA pathways presented in this article. PMID- 11396095 TI - Pathogenesis of rheumatoid arthritis. The role of T cells and other beasts. AB - The evidence coming from the different experimental approaches reviewed in this article strongly supports the hypothesis that RA is T-cell driven at all stages of the disease. Although the effector phases responsible for the events that lead to joint destruction involve several different cell types, cytokines, and other mediators, T cells still direct operations behind the scenes. Direct experimental proof of this proposition in patients is still lacking, but the development of nondepleting modulating CD4 monoclonal antibodies may provide new tools to test this hypothesis. In this respect, it is encouraging that using one such reagent, we have recently shown that not only did the activity of the disease improve but, more importantly, the inflammatory indices and production of non-T-cell cytokines were reduced. This is not to dissimilar from the results of experiments described in animals, where by blocking synovial T cells, the production of IL-1 beta and TNF alpha could be decreased by more than 90%. From this perspective, it may be predicted that by modulating T cells in the joint, it is possible to achieve our ultimate goal of permanently switching off the disease. PMID- 11396096 TI - Pathogenesis of rheumatoid arthritis. Role of B lymphocytes. AB - Despite many years of investigation, there remain many unanswered fundamental questions on the role of B cells in RA. Why is RF found in the sera of 80% of patients with RA and often in other chronic inflammatory diseases? What signals lead B lymphocytes to migrate into the subsynovial lining of joints? Does receptor revision in synovium play a role in the generation of autoantibodies in RA? What is the relative contribution of B-cell inhibition on the salutary effect of medications for RA? Can targeting autoreactive B cells, in conjunction with other therapies, provide therapeutic benefit in RA? We are hopeful that through continued basic, clinical, and translational research, these questions can be answered. PMID- 11396097 TI - Pathogenesis of rheumatoid arthritis: the role of synoviocytes. AB - Considering the characteristics of RA synovial tissues such as marked proliferation and invasion to adjacent tissues, comparisons with transformed or neoplastic tissue are natural. RA synovial tissues or cells are not truly malignant, but they have many features of transformation, denoted as "partial transformation" in this article. These features include anchorage-independent growth, loss of contact inhibition, oncogene activation, monoclonal or oligoclonal expansion, detectable telomerase activity, and somatic gene mutations. Although it is not possible to conclude whether most of these cells are permanently changed in association with some genetic alterations or are passively changed by virtue of environmental factors (i.e., cytokine-mediated imprinting), the presence of p53 mutations in RA synovial tissues is especially persuasive. A number of transcription factors play a critical role in the activation, differentiation, and proliferation of RA synovial cells. In particular, the roles of AP-1, MAPKs, and NF-kappa B have been investigated carefully because of their ability to regulate numerous inflammation-related genes. These transcription factors also control expression and activation of matrix-degrading enzymes, including MMPs, aggrecanase, and cysteine proteases, which are the primary enzymes responsible for joint destruction. Elucidation of gene mutations and detailed signal transduction pathways that are specific to RA as well as mechanisms of action of matrix-degrading enzymes may lead to development of a novel therapy for RA. Careful mapping of cytokine networks a decade ago led to groundbreaking advances in therapy. Similarly, methodical evaluation and prioritization of intracellular targets might provide the basis for therapeutic interventions. PMID- 11396098 TI - Rheumatoid arthritis. From bench to bedside. AB - In the last decade, basic science has unraveled in considerable detail the inflammatory and related processes ongoing in RA synovial membrane. This has translated to cytokine targeting therapies with some effect. How much more can be achieved? It may be argued that the order of improvement obtained thus far obviates further study. This ignores the potential to achieve a far greater magnitude of disease suppression. Our objective in innovating biologic therapies should now be routine achievement of American College of Rheumatology "70% responses" or disease remission and design of patient-specific therapy based on individual disease characteristics. We have not yet explored the potential contained in combination biologic approaches, particularly when different processes within the disease are targeted. Cocktails might target T cells, cytokines, and angiogenic factors, for example. These developments must also be seen in the context of the information soon to be available from the Human Genome Project. The impact of gene array analysis and similar techniques that facilitate simultaneous evaluation of thousands of gene activation events is also awaited. This, in turn, is likely to require considerable development in our use of information technology, because the volume of information will soon (or may already) be prohibitive. Finally, encompassing genomic and bioinformatic approaches should certainly challenge our basic diagnostic criteria such that it ultimately may be necessary to consider our clinical diagnoses on the basis not only of clinical phenotype but of genotype and biologic response profile. Through this rapid evolution, close communication between physician and scientist is essential. PMID- 11396099 TI - Systemic low-dose glucocorticoid treatment in rheumatoid arthritis. AB - Glucocorticoids provide a large, immediate improvement in the symptoms of rheumatoid arthritis. At doses acceptable for long-term treatment, however, symptoms gradually re-emerge. Relatively low doses of glucocorticoids can, for several years, substantially retard the rate of joint destruction shown on radiographs. This differential effect implies the coexistence of two pathologic processes within diseased joints. Long term, low dose glucocorticoid therapy probably increases the risk of serious adverse effects, but an evidence-based case can be made for the limited use of low dose glucocorticoid treatment in early disease. PMID- 11396100 TI - Rheumatoid arthritis. Treatment of early disease. AB - Treatment of rheumatoid arthritis (RA) is changing with the rapid increase of information on the efficacy of old and new antirheumatic agents. This article provides an opinionated review of the data currently available and makes a case for rapid and aggressive treatment of all patients with RA as early as possible. In the future, corticosteroids will be an important component of such treatment strategies. PMID- 11396101 TI - Combinations of conventional disease-modifying antirheumatic drugs. AB - During the last decade, numerous new treatments and treatment approaches have literally transformed current thinking about rheumatoid arthritis (RA) and, more importantly, about the way patients are treated. The nearly universal acceptance of the use of combinations of disease-modifying antirheumatic drugs (DMARDs) to more effectively treat a growing percentage of patients with RA has been a central theme. Importantly, controlled studies have now shown many combinations to be well tolerated and significantly more effective than mono-DMARD therapy. PMID- 11396102 TI - Tumor necrosis factor-alpha blockade in the treatment of rheumatoid arthritis. AB - The use of TNF alpha antagonists in RA has been extremely instructive. They have taught us that selective targeting of a pathogenic element can provide substantial clinical benefit. They have reinforced the concept of TNF alpha as a pivotal cytokine in the pathogenesis of RA. Pharmacodynamic studies of TNF alpha antagonists have further clarified the pathogenic processes involved in the disease. TNF alpha antagonists have set a new therapeutic standard for RA. Indeed, they are one of the most important advances in the history of the treatment of the disorder. If clinical efficacy is sustained and the safety profile remains benign over the long term, TNF alpha antagonists may replace MTX as the gold standard and become the agent of choice for combination therapy in RA. Further studies are needed to clarify their ultimate position in the therapeutic algorithm. PMID- 11396103 TI - Potential biologic agents for treating rheumatoid arthritis. AB - The encouraging clinical results observed in trials using anti-TNF therapy clearly warrant further studies to determine whether TNF inhibitors are capable of modifying the destructive component of this disease in long-term follow-up studies as well as to assess the safety of long-term use (see the article by Keystone in this issue). It is also reasonable to propose that interfering with the cytokine cascade earlier in the course of disease may be of even greater therapeutic benefit. As the pathogenetic mechanisms in RA are more clearly defined, especially in early disease and in those individuals destined to develop severe disease, the potential of other biologic agents to specifically inhibit these critical pathways may provide better treatments for our patients. Many potential targets in the immune-mediated process of RA are currently being rigorously evaluated in clinical trials. Use of combinations of biologic therapies, perhaps in human patients with RA, should be of considerable interest in future trials. PMID- 11396104 TI - [Reflections on pediatrics in France and the United States: an appeal for comparative studies]. PMID- 11396105 TI - [Pediatric hospitalization: specifics and costs]. PMID- 11396106 TI - [Neonatal screening for sickle cell anemia: evaluation of a five-year experience in an area of northern Paris]. AB - We report a five-year experience of targeted neonatal screening for sickle cell disease in the northern part of the Paris area as well as the follow-up procedure of screened patients. POPULATION: This geographic area in France is characterized by a high frequency of populations at risk for sickle cell disease. RESULTS: Among 115,480 tested newborns, 250 patients were diagnosed (frequency: 1/462). The quality of the screening was attested by the high frequency (5.34%) of newborn carriers for a hemoglobin abnormality (n = 6168). We developed an optimized strategy which avoids the majority of pitfalls (false positive and false negative responses), except for S/HPFH. More than 95% of sickle cell disease was followed, the majority by medical sickle cell disease experts from hospitals. Only two deaths were recorded during this time period. CONCLUSION: We demonstrate the efficiency of targeting the proposed methodological strategy and the follow-up of affected newborns, a major argument demonstrating the importance of newborn screening. PMID- 11396107 TI - [Reduction of complimentary prescription examinations in a pediatric emergency department]. AB - Numerous biological tests and X-rays are prescribed though the patient's clinical status does not warrant them. They are a source of extra work, extra costs, and painful and potential measurement errors. OBJECTIVES: To reduce unnecessary tests in four frequent situations (bronchiolitis, gastroenteritis, unexplained fever in children more than three months of age, asthma attack) at the pediatric emergency department of the Robert-Debre hospital (Paris). METHODS: Using a 'before/after' intervention survey, the intervention consisted of a modification of the request forms by adding prescription recommendations. All the children presenting with any one of the four targeted pathologies were included during the two 2-week periods. RESULTS: Test requests were reduced from 0.98 test per child to 0.77 (p = 0.02). The percentage of necessary tests increased from 33 to 50% (p < 0.0001); the percentage of children who underwent unnecessary blood samples decreased from 30 to 15% (p = 0.01). At the same time, the percentage of tests which were appropriate but not prescribed did not increase during the intervention period. CONCLUSION: It is possible to reduce unnecessary tests in an emergency pediatric department by adding recommendations to the request form. Unfortunately, the effects of such intervention do not last if the recommendations are not regularly recalled. PMID- 11396108 TI - [Pneumatic dilatation of caustic esophageal stenosis in children. Report of five cases]. AB - The ingestion of caustic substances is a frequent accident in the child. The treatment of caustic strictures of the esophagus, which constitute the most frightening complication, varies from conservative treatment to esophageal replacement. We report the preliminary results of a prospective work established since July 1998 and aimed at evaluating the role of pneumatic dilatation in the treatment of these injuries. STUDIED POPULATION: The series included five boys with an average age of six years. Two children were neurologically handicapped. RESULTS: The caustic product was soda (three cases), a dishwasher solution (one case) and bleach (one case). Dysphagia was constant and concerned the semi-fluids in four cases. The stricture was extended over an average length of 4 cm. The total number of dilatations was 36, with an average of seven sessions/patient. No complication was noted. The result was good in four cases. The only failure was noted in a child who presented a gastroesophageal reflux secondary to the scarring phenomena. CONCLUSION: Pneumatic dilatation constitute a therapeutic mean whose results are attested. The best treatment remains, however, preventive: it is the setting out of reach of the child the dangerous products. PMID- 11396109 TI - [Analysis of delay in diagnosis of extrahepatic biliary atresia]. AB - Prognosis of extrahepatic biliary atresia depends on an early surgical treatment. This survey studied the delay to surgery of infants treated for biliary atresia and analysed the causes of late diagnosis and referral. MATERIAL AND METHODS: Medical files of 21 infants treated for biliary atresia between 1988 and 1998 were retrospectively analysed. RESULTS: Median age at biliary operation was 57 days and did not change during time. In only 3/21 cases, surgery was performed before 45 days of age. The first clinical or biological sign of cholestasis was noted at a median age of 12 days. In eight cases, the first medical visit for cholestasis (median age of 21 days) was not followed by a blood test. In 11 cases, infants presenting with biological cholestasis were referred to hospital more than seven days later. False diagnosis was noted in seven cases and delayed significantly the operation. CONCLUSION: The delay to surgical treatment is too long and does not decrease whereas a majority of infants visit their physician early. Medical information is mandatory for all physicians taking care of infants. PMID- 11396110 TI - [Dermoid cysts revealed by meningitis with medullary compression]. AB - The spinal dermal sinus tracts in the lumbosacral region are not usually recognized, especially when they are not associated with other cutaneous lesions. In these sites, the sinus tracts communicate with the dura in 90% of cases, leading to an important risk of meningitis. CASE REPORTS: Two infants (9 and 12 months old) were hospitalized for meningitis. The hospitalization was preceded two weeks earlier by stubborn constipation, which revealed a neural compression. The physical and MRI examination showed a lumbosacral sinus in contact with a dermoid or an epidermoid tumor. These cysts were infected with anaerobic organisms. Despite surgery and antibiotic therapy, one child remained paraplegic. CONCLUSION: Dermal sinuses above the intergluteal crease should be surgically excised at the time of diagnosis in all patients, regardless of the patient's age or neurologic findings. PMID- 11396111 TI - [Meconium periorchitis. Two case reports]. AB - CASE REPORTS: Two cases of meconium periorchitis respectively in a ten-day- and a five-month-old infant are reported. The presenting symptom was a scrotal mass, isolated in one case and associated with pain and inflammation in the other case. Intraoperative histological diagnosis allowed conservative surgery in one case. COMMENTS: About 40 cases of scrotal manifestations of an intrauterine perforation of the gastrointestinal tract are reported in the literature. In cases with associated intraperitoneal calcifications, the diagnosis is easy and surgical exploration may be avoided, spontaneous resolution being the rule. However, in half of the cases, a scrotal mass is the only manifestation and raises the question of a testicular tumor. CONCLUSION: Unfamiliarity with this unusual entity may lead to unnecessary orchiectomy. PMID- 11396112 TI - [Medullary arteriovenous malformations in children]. AB - Spinal arteriovenous malformations are rare lesions in children but may produce serious functional disorders and residual handicaps. CASE REPORT: We report the case of a 10-year-old patient who had weakness in his left leg with intermittent urinary incontinence. Neurological examination showed a spastic paraparesis of both legs, more obvious in the left leg. MRI led to the diagnosis of a spinal vascular malformation, which was confirmed by angiography. The patient underwent a session of embolization, which resulted in a significant clinical improvement. CONCLUSION: The importance of MRI in such a case is emphasized as is therapeutic management with embolization. PMID- 11396113 TI - [Bullous cutaneous mastocytosis revealed by acute disease secondary to morphine administration]. AB - Mastocytosis in children shows in three clinical forms. The rarest is the diffuse or bullous form. CASE REPORT: Since one month of age an infant showed a diffuse erythema and vesicle rash. At four months of age, serious discomfort after morphinic absorption led to the diagnosis of bullous cutaneous mastocytosis. Histologic examination confirmed this diagnosis. The clinical severity led to intravenous corticosteroid and antihistamine therapy. COMMENTS: Bullous cutaneous mastocytosis is unusual in children. However, it should be considered if there are any doubts because of its serious complications and iatrogenic therapeutic risks. Some serious cases require intravenous corticosteroid therapy. Appropriate care enables a normal development with a disappearance of the disease before the teenage years. PMID- 11396114 TI - [Calcium-sensing receptors: physiology and pathology]. AB - The near constancy of extracellular calcium concentration is required for the numerous physiological functions of extra- and intracellular calcium. This implies that any change in extracellular calcium concentration must be detected in order to allow the appropriate correction by the homeostatic systems. The identification and cloning of a calcium-sensing receptor (CaR), which is expressed in the plasma membrane of parathyroid cells as well as many other cell types, has been a major advance in the understanding of the mechanisms involved in the control of extracellular calcium concentration. In addition, it demonstrated that extracellular calcium concentration itself is the first informative hormone-like messenger in this system. CaR belongs to the C subfamily of seven transmembrane-spanning G protein-coupled receptors. Several inherited disorders in extracellular calcium homeostasis are due to both activating or inactivating mutations in CaR gene, strengthening the essential role of CaR in the control of calcium metabolism. PMID- 11396115 TI - [Congenital long QT syndrome]. AB - Long QT syndrome (LQTS) is a clinically and genetically heterogenous syndrome characterized by a lengthening of the QT interval on the surface ECG and a propensity to severe ventricular arrhythmias such as torsades de pointes and ventricular fibrillation, leading eventually to syncope and sudden death. This rare syndrome with a mendelian inheritance occurs in subjects with otherwise normal cardiac morphological examination. The potentially severe prognosis justifies a presymptomatic diagnosis. The genetic nature of the disease has been confirmed with the identification of at least six loci and five genes. This syndrome is a perfect illustration of an adrenergic-induced ventricular arrhythmia. The first-line treatment is a beta-blocking agent for all symptomatic patients. In addition, a number of drugs known to lengthen ventricular repolarization must be prohibited. In case of suspicion of LQTS, all family members should be tested both clinically with a surface ECG and genetically in order to diagnose presymptomatic patients. PMID- 11396116 TI - [Differential diagnosis of elevated transaminases]. AB - Elevated serum levels of transaminases must always be considered as an abnormal finding. Drugs and toxins must be eliminated first as possible hepatotoxic agents or co-factors. Antiviral hepatitis A IgM determination is the first test to perform. However, other viruses with spontaneous benign courses are the most frequent cause. Only if the initial presentation is severe, or if liver tests remain abnormal after several weeks, can other rare diseases be sought. Liver biopsy is rarely necessary to reach a diagnosis. PMID- 11396117 TI - [The consequences of parental smoking in children: pediatricians must act]. AB - Prenatal and childhood passive tobacco smoke exposure resulting from parental smoking may have severe side effects, such as low birth weight, prematurity, sudden infant death syndrome, upper and lower respiratory tract infections and asthma. By giving information to parents, and particularly by emphasizing the dangers of passive smoke exposure for their children, pediatricians have a critical role to play in their prevention. This may also be helpful for adolescents who are starting to smoke actively by trying to understand the needs that they express by this behavior, and encouraging them to go to a stop smoking counseling center. PMID- 11396118 TI - [BREV: a new clinical scale for the evaluation of cognitive function in school age and preschool-age children]. AB - BREV (Batterie Rapide d'Evaluation des fonctions cognitives) is a new evaluation test for the screening of cognitive disorders in 4-9-year-old children, based on a neuropsychological process. It is made up of 17 subtests which have been carefully standardized. It is not an intelligence test but a tool for children' health professionals to use as a rapid neuropsychological screening test. It is particularly recommended for any child with a school learning disorder or neurological history with a high risk of cognitive disturbances such as epilepsy. It may also be used as a systematic screening test. PMID- 11396119 TI - [In situ fluorescent hybridization and trisomy 13 mosaicism: an example of the difficulty in managing genetic diseases in Togo]. PMID- 11396120 TI - [Gingival granular cell tumor in a newborn]. PMID- 11396121 TI - [Hair strangulation syndrome in children]. PMID- 11396122 TI - [Performance of Slovak hospitals as related to Porter's generic strategies]. AB - Porter's generic strategies characterize organizations in terms of their competitiveness, and are related to the performance of the organization. The aim of this study was to analyze the Porter's generic strategies and their effect on performance in the context of the Slovak hospital industry. Acute care hospitals with more than 30 beds were included into the study. National institutes providing specialized service were excluded from the study. Strategy and performance were evaluated on the basis of self-reported questionnaires, completed by chief administrators of hospitals (total 76 completed questionnaires were obtained, out of 81 distributed, i.e. 94% response rate). The cluster analysis was used for the identification of strategic orientation. Performance differences across strategic groups were tested using multivariate analysis of covariance (MANCOVA). The hierarchical cluster analysis uncovered a four-group taxonomy of hospitals: the group "Focused Cost Leadership" included 33% of hospitals, the group "Stuck-in-the middle" 49%, the group "Wait and See" 13% and the group "Cost leadership" 5%. Significant differences in performance were related to the Porter's pure, or hybrid strategies, respectively. In terms of industry evolution, the Slovak hospital industry could be characterized as fragmented, having a large number of small and medium size mainly state owned hospitals, with absence of market leaders, and with high exit barriers (mainly social and political) that hold back consolidation. (Tab. 1, Ref. 35.). PMID- 11396123 TI - [Assessment of functional capability of liver parenchyma using indocyanine green before liver resection]. AB - During the period from 1997 to 2000, the total of 129 liver operations were performed at the 2nd Surgical Department of the Masaryk's University in Brno. Malignant tumours were present in 91 cases. In-hospital mortality in patients operated on for malignant liver tumours was 2.1%. No case of death was caused by liver insufficiency. Examination of liver function by use of indocyanine green (ICG) enabling the determination of the extent of possible resection, contributes to the lowering of postoperative morbidity and mortality. The rate of indocyanine green retention up to 6% does not limit the extent of resection, the retention rate up to 15% allows bisegmentectomy or a minor surgery, the retention rate up to 20% allows segmentectomy at the most, and the retention rate up to 30% enables superficial excisions only. PMID- 11396124 TI - Hippocrates and his principles of medical ethics. AB - "The Father of Medicine", as Antiquity called Hippocrates has left rich medical and ethical heritage for us. His heritage--the collection of treatises Corpus Hippocraticum, from 5th and 4th centuries BC, comprise not only general medical prescriptions, descriptions of diseases, diagnoses, dietary recommendations etc., but also his opinion on professional ethics of a physician. The Hippocratic Oath, taken by ancient and medieval doctors, requires high ethical standards from medical doctors. Its principles are important in professional and ethical education of medical doctors even today. (Ref. 4.). PMID- 11396125 TI - Is it possible to influence the mortality in children with hemolytic uremic syndrome? AB - THE CURRENT STATE: Hemolytic uremic syndrome (HUS) is the most frequent cause of acute renal failure in children. In our geographic conditions D(+)HUS prevails, in its etiology E. coli O157:H7 is represented most. In the course of HUS there can occur extrarenal damage to some organs, the manifestation of multiple organ failure is then possible. Delayed diagnosis of HUS, its complicated course and therapeutical strategy influence greatly the mortality rate of affected children. SUBJECTIVE: The optimal therapeutical procedure was elaborated on the base of evaluating the causes of death in children with HUS, and, simultaneously, there were revealed further reserves that can decrease the mortality rate. METHODS AND MATERIAL: A retrospective analysis of mortality within 1982-2000 was carried out in one of three centres dealing with the therapy for HUS in children in the Czech Republic. The total number of HUS children was 69 (40 girls, 29 boys, mean age 4.5 years). 9 patients out of the analyzed group died. RESULTS: Out of 9 children (4 girls, 5 boys) died, D(+)HUS was present in 7 (77.8%, 4 girls, 3 boys, mean age 2.1 years), D(-)HUS occurred in 2 subjects (22.2%, boys, mean age 7.5 years). Dialysis therapy was needed in all 9 children (peritoneal dialysis and/or hemodialysis). The mortality rate in our group of children was 13%. An autopsy carried out in dead children showed dominant severe affection of kidneys/brain/heart. CONCLUSION: In spite of marked decrease in the number of HUS children died particularly in developed countries, permanent attention must be paid to this disease. Besides early diagnosis, corresponding therapy is necessary. That should be performed at a specialized centre. The elaborated algorithm can be used in the therapy for HUS. This is one of the ways for further decrease of both mortality and chronic morbidity in children suffering from HUS. (Tab. 3, Fig. 1, Ref. 15.). PMID- 11396126 TI - Immunohistochemical detection of LRP protein in the normal human lung. AB - In this study, we have determined the LRP (lung resistance-related protein) by immunohistochemical method. LRP belongs to proteins which cause the multidrug resistance (MDR). It has been found in various normal human tissues, where it plays a protective role against toxic compounds. Multidrug-resistant cells distribute the cytotoxic drug into the perinuclear region and then redistribute it back into the cytoplasm. It is just a hypothesis today that LRP can mediate drug resistance by regulating both the cytoplasmic redistribution and the nucleocytoplasmic transport of drugs. In order to detect LRP we have used the paraffin-embedded sections of the normal human lung tissue. LRP was predominantly located in two regions: 1) in bronchial epithelial cells and 2) in alveolar macrophages. Positive cells were coloured brown and showed strong reactivity. Negative control included the omitting of primary antibody and replacing it by buffer solution. Bronchial epithelial cells and alveolar macrophages stayed uncoloured, i.e. unreactive. (Fig. 5, Ref. 17.). PMID- 11396127 TI - Posterior scleroplasty in children with severe myopia. AB - PURPOSE: To assess the influence of scleral reinforcement on the evolution of severe myopia in children. MATERIAL AND METHODS: Scleral reinforcement after Thompson was performed on 251 eyes of 154 children with high myopia from 2 to 18 years of age. The main indication criteria for surgery during the period 1992 2000 included: severe myopia more than -7 Diopters and the increase in refraction more than -1 D(per year. Zenoderm (porcin skin) was the main alloplastic material used during surgery. No serious complications were observed. The following main indicators of myopia advancement were investigated on a long-term basis and evaluated: axial length, refraction, visual acuity, fundus findings and perimetry. RESULTS: The positive influence of surgery on myopia advancement was observed in 100% of patients. In about 53% of operated eyes, myopia was absolutely stopped, and in about 47% of operated eyes, its advancement was considerably reduced. During 10 years of postsurgical check-up, stabilisation of myopia was achieved, the following in individual indicators: axial length--53.8% of eyes, refraction--52.9% of eyes, visual acuity--85% of eyes, fundus findings- 58.6% of eyes, perimetry--59.1% of eyes. The advancement of myopia in other 47% of patients has been decreased from 1.1 D/per year before surgery to 0.1 D till 10 years after surgery. CONCLUSION: Scleral reinforcement is an effective and safe surgery that can stabilise the progression of severe myopia in children. (Tab. 6, Fig. 7, Ref. 12.). PMID- 11396128 TI - [Transurethral resection of the prostate in the treatment of benign prostatic hyperplasia]. AB - BACKGROUND AND OBJECTIVES: Transurethral resection of the prostate (TURP) represents a method of choice in surgical treatment of benign prostatic hyperplasia (BPH). The objective of this prospective study was to evaluate TURP mortality and morbidity rates. PATIENTS AND METHODS: In 1998 184 patients with lower urinary tract symptoms (LUTS) indicating the presence of BPH underwent one of the following surgeries: TURP 149 (81%), transurethral incision of the prostate 19 (10.3%), and open surgery (suprapubic transvesical prostatectomy) 16 (8.7%). From the group of 149 patients treated by TURP the following patients were withdrawn from the study: 7 (4.7%) patients with a finding of incidental carcinoma of the prostate and 14 (9.4%) patients who did not attend the postoperative follow-ups. RESULTS: 64 (50%) patients were treated for LUTS syndrome before TURP. TURP was applied in 69 (53.9%) cases for absolute indications. During the surgery and within one year after it, no mortality was recorded. Complications during the surgery were present in 13 (10.2%) patients, and within 24 h after the surgery in 38 (29.7%) patients. Early postoperative complications (up to 4 postoperative weeks) were recorded in 49 (38.3%) patients. Late postoperative complications within 3 month after the surgery occurred in 16 (12.5%) patients, within 6 months in 17 (13.3%) patients, and after 12 months in 17 (13.3%) patients. CONCLUSIONS: Although complications occur almost in 58% of patients, TURP still represents the standard treatment of LUTS indicating the presence of BPH. (Tab. 5, Fig. 2, Ref. 15.). PMID- 11396129 TI - The review of imported visceral leishmaniosis in the Czech Republic. AB - BACKGROUND: From the late 1950s through 2000, a total of 8 cases of imported visceral leishmaniosis (VL) were registered in the Czech republic. OBJECTIVES: The authors were made to point to the issue of imported VL by the fact 3 cases of this disease (imported from East Africa, Croatia, and southern Italy) were reported in 1999, plus another one (again imported from Croatia) in 2000. METHODS: The case reports of 4 cases of imported VL are presented. They are cases 5-8 ever reported in the Czech Republic. RESULTS: The infection manifested itself by fever, marked splenomegaly, leukopenia, thrombocytopenia, and rapid weight loss. The diagnosis was confirmed by the microscopic finding of amastigotes in punctate obtained from bone marrow, liver, spleen and, also, by serology. All the patients were successfully treated with amphotericin B. CONCLUSION: Infection by VL should be considered when establishing the diagnosis not only in patient returning from endemic regions and show hepatosplenomegaly, fever, leukopenia, and thrombocytopenia. Given the long incubation time, VL may be encountered also in foreigners who had lived in the above regions. Besides, the diagnosis of VL should also be considered in immunocompromised individuals. (Ref. 27.). PMID- 11396130 TI - Results of peripheral nerve reconstruction by autograft. AB - OBJECTIVES: This paper presents the results achieved in microtechnique surgeries performed during a 15-year-long period (1985-1999). By performing surgeries on 60 patients, 63 nerves were treated. MATERIAL AND METHODS: In 42 patients with injuries of peripheral nerves of upper extremities, 45 nerves were reconstructed by autografts. 14 patients were subjected to reconstructive surgeries on peripheral nerves of lower extremities. In 4 patients we reconstructed the facial nerve by means of autograft. The analysis of surgical effects has been made in dependence on indicators as follows: period elapsed from injury to surgery, age of patient, nature of injury, length of autograft, location of injury, kind of nerve inflicted. RESULTS: When assessing the results of reconstructive surgeries of peripheral nerves of lower and upper extremities we observed a big difference on the behalf of upper extremities. High efficiency can be seen in tibial nerve surgeries of lower extremities. In general we achieved good results in facial nerve reconstructions. CONCLUSION: The crucial factor that has an impact on the result of surgery is that of the time which elapsed from injury to reconstructive surgery. The factor is especially marked in younger patients. (Tab. 7, Ref. 15.). PMID- 11396131 TI - Strategy-making capability and performance of Slovak acute hospitals. AB - BACKGROUND: Research of strategic management issues in the environment of Slovak Health Care System generally and in the environment of secondary health care providers partially is a relatively new approach of last decade. In continuity with research of western authors this pilot study approaches a specific area of strategic management that is strategy-making capability and its relationship to performance of hospitals. AIM: Main goal of authors was to apply framework developed by Hart and test relationship of strategy making capability and performance in Slovak Acute Hospitals. Two hypotheses were developed for that purpose. METHODS: To conduct a test of both hypotheses a combination of three instruments was used. First was a set of 17 questionnaire items to tap the five strategy-making modes. Second was another set of 8 questionnaire items to tap the four perceived performance dimensions. Third was a current classification of Slovak acute hospitals and their levels. In summary data on twenty-five questionnaire--items were mailed to Chief Executive Officers (CEO) of 81 Slovakian acute hospitals within a larger survey co-ordinated by Ministry of Health. Data were collected via a questionnaire survey of top managers from selected acute Slovak hospitals within December 1999--February 2000. The sample was chosen from the list of acute care hospitals in the 1999 Report of the Slovak Institute for Health Care Information and Statistics. RESULTS: Being aware of certain limitations in such a construct results indicate that hospitals with higher level of strategy-making capabilities might outperform those with lower level of this capability. (Tab. 5, Ref. 21.). PMID- 11396132 TI - Relations between the expression of p53, c-erbB-2, Ki-67 and HPV infection in cervical carcinomas and cervical dysplasias. AB - BACKGROUND: The analysis of mutual relations between HPV infection and the expression of cancer gene products and proliferative activity in cervical carcinomas and dysplasias. MATERIALS AND METHODS: The expression of p53, c-erbB-2 oncoproteins and proliferative activity (Ki-67) was evaluated immunohistochemically in 41 cervical carcinomas and 29 dysplasias. HPV infection (type 16, 18) was assessed by in situ hybridization technique. RESULTS: HPV positive carcinomas were found in 68.3% of cases. HPV type 16 infection were detected in 54% and HPV 18 in 39% of carcinomas. Simultaneous appearance of both virus types was shown in 25% of carcinomas. In dysplastic lesions, HPV infection was observed in 62.1% of cases. HPV type 16 was found in 34.5% and HPV 18 in 44.8% of patients. Both virus types were found in 17.2% of dysplasias. HPV infection was more extensive in cervical carcinomas than in dysplasias. Similarly the expression of oncoproteins was more intensive and referred to a higher percentage of cells in carcinomas. No relations between p53, c-erbB-2 overexpression and HPV infection were found. Ki-67 activity was found in a higher percentage of HPV-positive than in HPV-negative, both carcinomas and dysplasias. CONCLUSIONS: HPV infection, especially accompanied by increase of proliferative activity in dysplasias may define the cell subpopulation predisposed to malignant process development. The employment of in situ hybridization technique appears to be useful in detecting the viral infection in cytological smears even with no morphological changes in the cells. PMID- 11396133 TI - Plant polysaccharide PSK: cytostatic effects on growth and invasion; modulating effect on the expression of HLA and adhesion molecules on human gastric and colonic tumor cell surface. AB - PSK is a plant polysaccharide widely used for cancer immunotherapy in Japan and other Asian countries. It is considered that its antitumor effect is derived from its immunomodulating activity on the tumor-bearing host. The present study was designed to assess the direct action of PSK on in vitro proliferation and invasion of human KATO-3 gastric and Colo205 colonic cancer cell lines, and the expression of surface molecules such as HLA and adhesion molecules on these cells. The in vitro growth of KATO-3 cells was significantly inhibited by 100 micrograms/ml of PSK 48 hrs after culture initiation, and that of Colo205 was significantly inhibited by 10 and 100 micrograms/ml of PSK 24 hrs after culture initiation. The effect of PSK on the in vitro invasion of the tumor cells, assessed with a Matrigel invasion chamber, revealed that invasion of KATO-3 and Colo205 cells was inhibited by more than 10 micrograms/ml and more than 5 micrograms/ml of PSK, respectively. KATO-3 cells expressed HLA-ABC, HLA-A2/A28, HLA-DR very weakly, at almost baseline levels, but HLA-B27, B2-microglobulin and HLA-DQ were expressed at various levels. After treatment of KATO-3 cells with PSK, the expression of HLA-B27 and beta 2-microglobulin was significantly enhanced. Colo205 cells expressed all class-I antigens tested in this study at different levels, but class-II antigens at almost baseline levels. PSK also enhanced the expression of class-I antigens on Colo205 cells. ICAM-1 was expressed on KATO-3, but not on Colo205. The expression of ICAM-1 was enhanced to a greater extent by treatment with 10 micrograms/ml than with 100 micrograms/ml of PSK. Adenocarcinoma antigen AC-81 was strongly expressed on both cell lines, but PSK-treatment significantly enhanced its expression. These results suggested that enhancement of HLA class-I expression on tumor cells after PSK treatment may be one of the mechanisms responsible for the induction of anti-tumor immunity by PSK. PMID- 11396134 TI - Expression of heat shock proteins HSP70 and HSP27 in primary non-small cell lung carcinomas. An immunohistochemical study. AB - The present study was undertaken to determine the expression pattern of the hsp70 and the hsp27 genes in 106 cases of non-small cell lung carcinoma (NSCLC). We have shown that in the majority of cases (95/106) the HSP70 immunoreactivity was localized both in the cytoplasm and the nuclei. We also observed an enhanced nuclear immunoreaction for HSP70 in dysplastic lesions and in stage I tumors. In the case of the HSP27 we found a positive cytoplasmic immunostaining in 70% of cases, with the highest score in squamous cell carcinoma (SCC). We noted a positive correlation between the expression level of HSP27 and HSP70. There was a correlation between Ki-67 proliferation index and nuclear HSP70 staining, but not for HSP27. No association between the HSPs and the expression of pRb p16 and p21WAF1/CIP1 and p53 was found as studied previously. An interesting and statistically significant relationship was found between the expression of cyclin D1 and high intensity of HSP27 and HSP70 immunostaining. The relation of our results concerning the expression pattern of the HSP70 and HSP27 in NSCLC to those obtained by others for different types of primary tumors is discussed. PMID- 11396135 TI - Modulation of cancer cell multidrug resistance by an extract of Ficus citrifolia. AB - Multidrug resistance due to P-glycoprotein is a serious impediment to successful chemotherapy of cancer. Previous studies have shown that natural compounds such as prenyl flavonoids are able to modulate the multidrug resistance phenotype of P glycoprotein-positive cancer cells. A fraction from the dichloromethane extract of a common Guadalupe Ficus, Ficus citrifolia was studied for its direct interaction with the purified C-terminal cytosolic domain of P-glycoprotein, and for its induced accumulation and cytotoxicity of vinblastine and daunomycin in two model cell lines overexpressing P-glycoprotein, namely K562/R7 and MESSA/Dx5. The fraction bound with high affinity to P-glycoprotein C-terminal cytosolic domain and was as efficient as cyclosporin A to increase intracellular accumulation of daunomycin in K562/R7 leukemic cells. Moreover, the fraction markedly enhanced the cytotoxic effect of vinblastine on the growth of MESSA/Dx5 cells. These results suggest that Ficus citrifolia possesses important therapeutic potential for improving the efficacy of cancer chemotherapy. PMID- 11396136 TI - Analysis of cellular sensitization with cisplatin-induced apoptosis by glucose starved stress in cisplatin-sensitive and -resistant A431 cell line. AB - BACKGROUND: Most solid tumors show resistance to current chemotherapy. This drug resistance can be associated with stress conditions, such as glucose starvation, low pH and hypoxia. These conditions are not observed in normal tissues. These tumor specific conditions commonly cause the glucose-regulated stress response of cancer cells. This stress response leads to induction of resistance to multiple anticancer drugs, such as etoposide, doxorubicin, camptothecin and vincristine. An examination was made of the effect of the glucose-regulated protein (GRP) inducing conditions on cellular sensitivity to cisplatin (CDDP), which is a widely used drug against solid tumors. MATERIALS AND METHODS: We used CDDP resistant subline A431/CDDP2 from human epidermoid carcinoma cell line A431 as previously established. Glucose-starved response showed GRP78 protein by Western blotting. We generated the glucose-starved condition by exposing cells to 2 deoxyglucose (2-DG) and examined cellular sensitivity to CDDP. We next examined CDDP induced apoptosis in glucose-starved condition. Furthermore, we determined the JNK1/SAPK and caspase-3 protein levels by Western blotting. RESULTS: The glucose-starved condition was up-regulate GRP78 in the parent A431 cell (A431/P). On the other hand, in A431/CDDP2, GRP78 levels were not changed. The GRP78 inducing stress condition led to cellular sensitization to CDDP in A431/P cells only. DNA fragmentation of A431/P treated with 2-DG increased CDDP-induced apoptosis. A431/CDDP2 was not different with CDDP induced apoptosis in glucose starved stress condition. The reduction of CDDP-induced apoptosis under glucose starved stress condition was influenced by JNK1/SAPK and caspase-3 proteins. CONCLUSION: These results indicated that the glucose-starved stress condition was associated with increased sensitivity to CDDP in the up-regulated GRP78 cells. Furthermore, the reduction of CDDP-induced apoptosis can be considered as one of the CDDP resistant mechanisms in a glucose-starvation condition. PMID- 11396137 TI - Selective activity of BS-RNase against anaplastic thyroid cancer. AB - BACKGROUND: Anaplastic thyroid carcinoma is an aggressive solid tumor that fails to adequately respond to any known chemotherapeutic regimen. The development of effective chemotherapy agents would provide the best chance for long-term survival of patients. MATERIALS AND METHODS: The cytotoxic effects of bovine seminal ribonuclease (BS-RNase) against thyroid carcinoma cell lines with different degrees of differentiation in comparison to non-malignant cells, including human foreskin fibroblasts (HFF) and retinal pigment epithelial cells (RPE), were tested using the MTT dye reduction assay. Induction of apoptosis was demonstrated by annexin V assay and expression of proteins related to apoptosis was investigated by flow cytometry. The antitumoral in vivo effects of BS-RNase were assessed on established xenografts of anaplastic thyroid carcinoma cell line 8505C in nude mice using subcutaneous injections of BS-RNase (12.5 mg/kg once a day, on 20 consecutive days). RESULTS: All the tumor cell lines exhibited marked sensitivity against BS-RNase in comparison to HFF and RPE cells. The greatest growth inhibition was seen in the 8505C line, while IC50 values for papillary (B CPAP) and poorly-differentiated thyroid carcinoma cells were about 6-fold higher. The cytotoxic action of BS-RNase was associated with induction of apoptosis. Expressions of Fas and Fas-ligand were not influenced by BS-RNase completely, while the down-regulation of Bcl-2 in treated cells was observed. In vivo treatment induced significant tumor regression after the course of 20 consecutive days. No apparent toxic effects of BS-RNase toward non-malignant cells were observed during the in vivo treatment. After cessation of therapy (day 20) tumor volume continued to decrease and the tumor was no longer detectable after 30 days of treatment induction in all animals. CONCLUSION: BS-RNase may have beneficial effects for treatment of aggressive anaplastic thyroid cancer. PMID- 11396138 TI - Expression and possible role of cyclin D3 in human pancreatic adenocarcinoma. AB - BACKGROUND: Cyclin D3, one of the D type cyclins, has been known as a cell cycle modulator at the G1-S phase. In the present study, we investigated its expression in pancreatic adenocarcinoma in order to elucidate its clinical role. MATERIALS AND METHODS: We examined cyclin D3 as well as, cyclin D1 overexpression in 60 pancreatic adenocarcinoma by means of immunohistochemistry. RESULTS: Cyclin D3 overexpression was found in 33.3% (20 cases) of the cases, being more frequently seen in cases showing high Ki-67 labeling index, stage IV, moderate- or poor differentiation, lymph node metastasis and large size and cyclin D1 overexpression. Multivariate logistic analysis independently linked cyclin D3 overexpression to tumor size (p = 0.0392) and stage (p = 0.0312). Cyclin D1 overexpression was observed in 41.7% (25 cases) and was related to Ki-67 labeling index, cyclin A overexpression, stage, carcinoma differentiation, lymph node metastasis, tumor size, lymphatic invasion, perineural invasion and retropancreatic invasion. It was independently linked to carcinoma differentiation (p = 0.0081), lymph node metastasis (p = 0.0426) and lymphatic invasion (p = 0.0269). CONCLUSION: These results suggest that cyclin D3, as well as cyclin D1, plays a role in the progression, including cell proliferating activity, of pancreatic adenocarcinoma. PMID- 11396139 TI - Sodium butyrate induces growth inhibition and modulates galectin-8 expression in human lung carcinoma cells. AB - Galectins are animal lectins, which may play a role in neoplastic transformation. Po66-Carbohydrate Binding Protein (Po66-CBP) belongs to the galectin-8 family and is expressed in lung tumor cells but not in normal ones. Recent studies showed that galectin-8 could be used for human lung squamous cell carcinoma radioimmunotherapy. To optimize this method of treatment, we attempted to increase galectin-8 expression in human lung tumor cells. A human lung squamous (SK-MES-1) or adeno (A 549) carcinoma cell line was grown with or without sodium butyrate. Cell growth, morphology, transcriptional, expression translational expression and cellular localization of galectin-8 were studied. 3 mM of sodium butyrate inhibited the two cell lines' growth after 48 hours of treatment, but only in SK-MES-1 cells galectin-8 expression is modulated without any secretion and cellular localization modifications, apoptosis or necrosis. Sodium butyrate could be an interesting tool in optimizing the radioimmunotherapy of human lung squamous carcinoma, but not of adenocarcinoma. PMID- 11396140 TI - Correlation of telomerase activity, clinical prognosis and therapy in oral carcinogenesis. AB - Telomerase activity is associated with most malignant tumors. To evaluate the role of telomerase reactivation as a diagnostic and prognostic marker in oral carcinogenesis activity was investigated in mortal and immortal cell lines in eight oral leukoplakias (OL) and 46 oral squamous cell carcinomas (OSCC). Activity was also investigated in 13 histopathologically unaffected mucosas from distant sites or tumor-free margins of the same patients using a modified, highly sensitive, non-radioactive telomeric repeat amplification protocol (TRAP). This was correlated with histopathological stages and the clinical course of the disease. 50% of OL and 46% of OSCC showed activity. One patient with positive, high dysplastic OL developed an OSCC 11 month later. One of three specimens of adjacent tissue presented activity and a recurrence occurred after 6 months. Out of 10 tissues of distal normal mucosa, 2 demonstrated activity which could also be proved in the corresponding tumor. Detection of telomerase reactivation may be a novel method for early detection of immortalised cell clones and malignant cells in histopathologically normal oral squamous epithelium. PMID- 11396141 TI - Oral shark cartilage does not abolish carcinogenesis but delays tumor progression in a murine model. AB - BACKGROUND: Shark cartilage and shark cartilage extracts have been reported to have anti-angiogenic and anti-neoplastic properties. This study reports the effects of oral administration of powdered shark cartilage on tumor progression in a murine renal tumor model. MATERIALS AND METHODS: Renal tumors were induced in CBA female mice by a single bolus of IV streptozotocin. 57 mice were fed shark cartilage and the numbers and rate of development of dysplastic convoluted tubules, papillary and solid renal epithelial tumors was compared with 57 control mice over an 88 week follow-up period. RESULTS: In the shark cartilage fed group dysplasia was first observed after 23 weeks (control 19 weeks), papillary tumors after 24 weeks (control 23 weeks) and solid tumors after 55 weeks (control 19 weeks). There was no significant difference in the rate of development of dysplastic tubules between test and control animals. The development of papillary and solid tumors was significantly delayed in the test group. CONCLUSIONS: In this tumor model oral shark cartilage delays, but does not abolish, tumor progression. PMID- 11396142 TI - Loss of heterozygosity of gene THW is frequently found in melanoma metastases. AB - We have recently described a new member of the PMP22/gas3 family of plasma membrane proteins referred to as THW. This gene is located on chromosome 6q and preliminary data have indicated a possible tumor suppressor gene function. We have therefore investigated LOH for gene THW in a panel of cancer cell lines and in a series of primary human melanomas as well as in melanoma metastases. We have detected LOH for gene THW in cell lines derived from melanoma, breast, pancreas, cervical, prostate and colon carcinoma with different prevalence, whereas the ovary carcinoma cell lines (n = 3) were negative. For melanomas we found a prevalence of LOH for gene THW of 10-20% in primary tumors, whereas in melanoma metastases we found a score of 50%. These data and the fact that the recently identified murine homologue PERP of gene THW mediates cell death in murine fibroblasts support the possible tumor suppressor function of gene THW. PMID- 11396143 TI - Cytotoxic activity of 5-benzoylimidazole and related compounds against human oral tumor cell lines. AB - A total of 24 benzoylimidazoles and structurally-related compounds were investigated for their cytotoxic activity against oral tumor cells and normal gingival fibroblast. Compound 23 (5-(2-hydroxylbenzoyl)-2-phenylimidazole) showed the highest cytotoxic activity against both human oral tumor cell lines (human squamous cell carcinoma HSC-2, human salivary gland tumor HSG) and normal human gingival fibroblast (HGF). Compounds 7 (2-(2-hydroxybenzoyl)benz imidazo[2,1 b]thiazole), 14 (1,3-diethyl-5-(2-hydroxybenzoyl)-4-imidazoline-2-thione) and 18 (5-(2-hydroxy-4-methoxybenzoyl)-3-methyl-2-methylimino-4-thiazoline) showed slightly lower cytotoxic activity, but higher tumor-specific cytotoxic action. The cytotoxic activity of compound 23 was significantly reduced by CuCl2, but not by CoCl2, FeCl3, or by antioxidants (N-acetyl-L-cysteine, sodium ascorbate, catalase). Compound 23 did not show any detectable oxidation potential (determined by NO monitor). Agarose gel electrophoresis demonstrated that compound 23 induced DNA fragmentation in human promyelocytic leukemia cells HL 60, but not in HSG cells. These data suggested that the response to compound 23 might be different from cell to cell. PMID- 11396144 TI - Effects of the butylated hydroxyanisole and butylated hydroxytoluene on the DNA adduct formation and arylamines N-acetyltransferase activity in human colon tumor cells. AB - The effects of butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT) on N-acetyltransferase (NAT) activity were examined using a human colon tumor cell line (colo 205). BHA or BHT were added to the cytosols or to the medium of human colon tumor cells: The NAT activity was measured by high performance liquid chromatography, assaying the amounts of acetylated 2-aminoflluorene (AAF), p aminobenzoic acid (N-Ac-PABA), nonacetylated 2 aminofluorene (AF) and p aminobenzoic acid (PABA). The NAT activity in the human colon tumor cells and cytosols was suppressed by BHA or BHT in a dose-dependent manner. The apparent values of Km and Vmax of NAT of human colon tumor cells were also decreased by BHA or BHT in cytosols and in intact cells. BHA or BHT may act as a noncompetitive inhibitor. After the incubation of human colon tumor cells with AF in the presence of BHA or BHT, the cells were recovered and DNA was prepared and hydrolysed to nucleotides. Adducted nucleotides were extracted into butanol and AF-DNA adducts were analysed by HPLC. The results also demonstrated that when BHA or BHT was added to the media, a decrease in AF-DNA adduct formation was seen in the human colon tumor cells. The finding of AF-DNA adduct formation in cultured human colon tumor cells suggest the possibility of using cultured cells for assessing arylamine-induced DNA damage. PMID- 11396145 TI - Induction of apoptosis in CTLL-2 cells by protocatechualdehyde. AB - The purpose of this study was to investigate the possible molecular mechanisms of the antiproliferative effect induced by protocatechualdehyde (PA, a dihydroxybenzene derivative). The viability of cytotoxic T cells (CfLL-2) stimulated by IL2 was significantly inhibited at 0.12 mM PA. This inhibitory effect was associated with the induction of apoptosis detected by DNA fragmentation assay. DNA ladder appeared at 0.12 mM PA and the intensity of DNA ladder was visible at 0.3 mM PA. PA inhibited the Ib2-dependent tyrosine phosphorylation of 91, 80 and 55 KDa proteins, but did not affect IL2-dependent serine/threonine phosphorylation of proteins. The levels of bcl-2 protein and mRNA were suppressed by PA. An alteration in bax protein expression on the apoptosis process in CTLL-2 cells was not observed. However, caspase-3 activity was increased by PA. Our results demonstrate that PA inhibited cell proliferation and induced apoptosis in CTLL-2 cells. It is concluded that PA is a potent anti proliferative agent and is expected to be a promising candidate for novel therapeutics. PMID- 11396146 TI - Alternative splicing of fibronectin transcripts in osteochondrogenic tumors. AB - BACKGROUND: Fibronectin (FN) acts in many fundamental biological processes including cell adhesion, migration, proliferation and apoptosis. Although FN shows an ubiquitous expression pattern, alternative mRNA splicing modulates tissue-specific molecular heterogeneity at three regions: extra domain-B, extra domain-A and variable region. MATERIALS AND METHODS: In the present study, we analyzed the FN mRNA splicing in osteochondrogenic tumors as well as in various normal tissues using the RT-PCR method. RESULTS: Normal cartilage almost exclusively contained short mRNA lacking alternative exons in all regions. However, 14 osteochondrogenic tumors including osteochondroma, enchondroma, chondroblastoma, chondrosarcoma and osteosarcoma uniformly contained long mRNA with various patterns of alternative exons in the three regions. CONCLUSION: These results indicated that the lack of regulation excluding the alternative exons may be associated with tumorigenesis in chondrocytes. PMID- 11396147 TI - Induction of apoptosis in an androgen-dependent mouse mammary carcinoma cell line by methylcobalamin. AB - SC-3 is a cloned cell line derived from an androgen-dependent mouse mammary tumor (Shionogi Carcinoma 115). A physiological level of androgen stimulates the growth of SC-3 cells through the production of androgen-induced growth factor. Methylcobalamin (MeCbl), one of the active cobalamins, inhibits the growth of SC 3 cells stimulated by androgen. It is known that apoptosis has an important role in tumor growth. The specific aim of this study is to examine the effects of MeCbl, in the presence of androgen, on apoptosis in SC-3 cells. Morphological analysis revealed budding nuclei and chromatin condensation in cells cultured with MeCbl, but few in cells cultured without MeCbl. Low molecular weight DNA extracted from cells cultured with or without MeCbl was analysed by gel electrophoresis. A characteristic nucleosomal size ladder was detected in the culture with MeCbl. The terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labelling method was also used to evaluate apoptotic cell death in SC-3 cells. Apoptosis was observed more frequently in SC-3 cells treated with MeCbl than in those without MeCbl. These results demonstrate that androgen-dependent SC-3 cells undergo apoptosis by MeCbl even if in the presence of androgen. PMID- 11396148 TI - Calretinin promoter for suicide gene expression in malignant mesothelioma. AB - To achieve specific and efficient expression of transfected suicide gene in malignant mesothelioma cells, we explored the 5-flanking region (2.2 kb) of the calretinin gene as a tumor-specific promoter. RT-PCR analysis demonstrated the expression of calretinin mRNA in two human malignant mesothelioma cell lines (H2052 and 211 H). A reporting vector containing the calretinin promoter (pCAL LUC) exhibited relatively high luciferase activity in these cell lines. After transfecting an expression vector containing the calretinin promoter-bound thymidine kinase gene (pCAL-TK) into the cells, we examined their sensitivity to ganciclovir (GCV). In the mesothelioma cell lines, the pCAL-TK transfected cells became about 100 times more sensitive to GCV than the parental cells. The calretinin promoter seems to be a promising candidate as a specific and efficient promoter in malignant mesothelioma. PMID- 11396149 TI - p53 evaluation in gastric mucosa of patients with chronic Helicobacter pylori infection. AB - Gastric cancer is often associated with p53 over-expression and Helicobacter pylori (HP) infection. In this study we have investigated the production of the p53 protein and mutation of its gene in precancerous gastric lesions with HP infection. For this purpose 130 patients who underwent endoscopy for dyspepsia were enrolled in the study. To assess p53 production and mutation of the p53 gene we employed an immunoluminometric assay and polymerase chain reaction single strand conformation polymorphism (PCR-SSCP) analysis, respectively. Histologically, 52 of the 130 enrolled patients showed intestinal metaplasia type I (IM) (90.4% of these were also HP positive), 47 had HP-related gastritis and 31 were normal. p53 cytosol levels were significantly higher in patients with IM or HP-related gastritis than in normal patients (p = 0.0137 and p = 0.0411, respectively). All DNAs extracted from gastric mucosa samples with higher p53 values and examined for p53 mutations by PCR-SSCP analysis were characterized by a normal run. Our data indicate, that irreversible genetic changes in the p53 protein has not yet occurred in morphologically non-neoplastic gastric mucosa with IM and HP-related chronic gastritis. In conclusion, the increase in p53 cytosolic levels found in our study is due to an increased production of the wild type protein probably related to an inflammatory response induced by HP infection. PMID- 11396150 TI - Paclitaxel immunoconjugate for the specific treatment of ovarian cancer in vitro. AB - A paclitaxel derivative was used for the preparation of a drug immunoconjugate for the specific targeting to ovarian cancer cells by the BCM43/2E5 monoclonal antibody. To determine the efficiency of conjugation we developed two reproducible immunoassays. Paclitaxel was detected on the conjugate by a paclitaxel immunoassay and the activity of the antibody was determined by a competition assay with BCM43/2E5 antibody. Paclitaxel-BCM43/2E5 immunoconjugate was tested on five human cell lines. This conjugate was cytotoxic for ovarian teratocarcinoma cells, ovarian adenocarcinoma and breast cancer cells, while the activity was negligible for normal cells. These results describe, for the first time, the synthesis of a soluble tumor specific- paclitaxel immunoconjugate which may establish a new mode for the use of this drug in cancer therapy for limiting side effects. PMID- 11396151 TI - Increased expression of peroxiredoxin II confers resistance to cisplatin. AB - Peroxiredoxin II (Prx II) has been known to be induced by various oxidative stimuli and to play an important protective role from oxidative damage by hydrogen peroxide (H2O2). In this study, we observed that cisplatin as well as H2O2 induced Prx II expression. To examine the correlation between the increased expression of Prx II and chemoresistance, we prepared a Prx II-overexpressing cell line, SNU638 cells, and found it to be more resistant to cell death induced by cisplatin and H2O2 than neo-transfectant cells. We also observed that enhanced expression of Prx II inhibited cisplatin- and H2O2-induced apoptosis, demonstrating that resistance to these cytotoxic agents was due to inhibition of apoptosis. The above results led us to suggest that the overexpressed Prx II protein inhibits cisplatin-induced apoptosis, thereby contributing to chemoresistance of tumor cells, especially to oxidative stress producing anticancer drugs. PMID- 11396152 TI - Detection of single-stranded DNA (ssDNA) in the vestibule of guinea pigs after the application of cisplatinum (CDDP). AB - BACKGROUND: Cisplatinum (CDDP) has a toxic effect on the inner ear. CDDP reacts with DNA and leads to apoptotic cell death. In this study, the presence of single stranded DNA (ssDNA) was examined immunohistochemically. MATERIALS AND METHODS: CDDP (10 mg/kg b.w.) was injected into the guinea pigs intra-peritoneally. Animals were sacrificed three days after the injection. The temporal bones were fixed via cardiac infusion. The tissues were embedded in paraffin and then used for immunohistochemical examination. RESULTS: That ssDNA was detected in the CDDP treated vestibule. The sensory epithelium, transitional area, dark cells and vestibular ganglion cells exhibited a positive immunoreactivity to ssDNA. CONCLUSIONS: DNA fragmentation is a characteristic feature of apoptosis. Fragmented DNA is detected as ssDNA. Our results indicate that CDDP involves the apoptosis in the inner ear and this is the cause of the ototoxicity of CDDP. PMID- 11396153 TI - Effects of retinoic acid and amifostine on in vitro growth of normal hemopoietic progenitor cells. AB - Association of all-trans retinoic acid (ATRA) and amifostine (AMF) might be an alternative in treatment of myelodisplatic syndromes. In this study we undertook a preliminary in vitro research on the effects of a combination of ATRA and AMF on normal hemopoietic progenitors. Mononuclear, non-adherent cells from peripheral blood of normal volunteers, were incubated with AMF, at a the concentration of 500 microM and then cultured for CFU-GM and BFU-E growth adding to culture dishes before gelling ATRA at the concentration 10(-6) and 10(-12)M. Controls were cultures not pre-incubated with AMF, cultures incubated with AMF without addition of ATRA and cultures without addition of any drug. ATRA addition did not have significant effects on hemopoietic progenitors growth. Association of ATRA and AMF do not appear to synergize to stimulate in vitro growth of hemopoietic progenitors. PMID- 11396154 TI - Metallothionein expression and apoptosis in pregnancy-dependent and -independent mouse mammary tumors. AB - BACKGROUND: A metal binding protein, metallothionein (MT), may be involved in the regulation of carcinogenesis and apoptosis in addition to various physiological processes. MATERIALS AND METHODS: MT and sex hormone receptors (estrogen and progesterone receptors, ER and PR) expressions, and apoptosis were investigated immunohistochemically in pregnancy-dependent (PDMT) and -independent mammary tumors (PIMT) in GR/A mice in order to evaluate the possible role of MT in the genesis and regression of tumors. RESULTS: In PDMT and PIMT, MT was localized in the nucleus and/or cytoplasm of tumor cells. In PDMT, MT expression and apoptotic figures were increased during the regression period after parturition. MT and ER expressions in PIMT were approximately the same as those in the growing PDMT, while PR expression was lower in PIMT than in the PDMT. A significant correlation was observed between MT expression and apoptosis in PDMT, but not in PIMT. CONCLUSION: The present study suggests that MT involvement in the PDMT regression is associated with apoptosis. PMID- 11396155 TI - Cytotoxicity of bioreductive drug tirapazamine is increased by application of electric pulses in SA-1 tumours in mice. AB - The application of electrical pulses (electroporation) is a local tumour treatment resulting in the facilitated accumulation of non-permeant chemotherapeutic drugs (electrochemotherapy), as well as in the transient reduction of tumour blood flow. The aim of our study was to determine whether the application of electric pulses to the tumour increased the antitumour effectiveness of the bioreductive drug tirapazamine (TPZ). The survival of SA-1 fibrosarcoma cells was 150-fold lower after the exposure of cells for 1 h to TPZ under anoxic compared with normoxic conditions. The exposure of cells to electric pulses did not increase the cytotoxicity of TPZ. However, the in vivo treatment of subcutaneous tumours with a combination of TPZ (i.p. 25 mg/kg) injected 20 min before the application of electrical pulses significantly enhanced tumour response. Treatment with TPZ and electric pulses, repeated three times at 24-hour intervals resulted in tumour growth delay of 7.2 days. The results of our study showed that the observed antitumour effectiveness is unlikely to be due to increased cellular accumulation of TPZ by application of electric pulses, as indicated from in vitro experiments. The effect is more likely to be attributed to increased tumour hypoxia as a consequence of reduced tumour blood flow induced by application of electric pulses. PMID- 11396156 TI - Urothelial papillary lesions. Development of a Bayesian Belief Network for diagnosis and grading. AB - The diagnosis and grading of urothelial papillary lesions are affected by uncertainties which arise from the fact that the knowledge of histopathology is expressed in descriptive linguistic terms, words and concepts. A Bayesian Belief Network (BBN) was used to reduce the problem of uncertainty in diagnostic clue assessment, while still considering the dependencies between elements in the reasoning sequence. A shallow network was designed and developed with an open tree topology, consisting of a root node containing four diagnostic alternatives (papilloma, papillary carcinoma grade 1, papillary carcinoma grade 2 and papillary carcinoma grade 3) and eight first-level descendant nodes for the diagnostic features. Six of these nodes were based on cell features and two on the architecture. The results obtained with prototypes of relative likelihood ratios showed that belief in the diagnostic alternatives is very high and that the network can identify papilloma and papillary carcinoma, including their grade, with certainty. In conclusion, a BBN applied to the diagnosis and grading of urothelial papillary lesions is a descriptive classifier which is readily implemented and allows the use of linguistic, fuzzy variables and the accumulation of evidence presented by diagnostic clues. PMID- 11396157 TI - Chemoprevention by difluoromethylornithine: correlation of an in vitro human cell assay with human clinical data for biomarker modulation. AB - The efficacy of difluoromethylornithine (DFMO) as a chemopreventive agent has been tested in vitro using a human epidermal cell (HEC) assay with growth inhibition and involucrin induction as endpoints. Suppression of polyamine content is currently being utilized as a biomarker in clinical trials for the chemopreventive efficacy of DFMO against colon cancer formation. We have now examined the effects of DFMO on suppression of polyamine content in the HEC assay. The findings indicate 1) the % change in spermidine to spermine ratio and the depletion of putrescine show excellent correlation with chemopreventive efficacy in vitro; 2) the effective concentrations in vitro overlap the plasma concentrations in the clinical trial. These observations serve as further validation of the usefulness of the HEC assay as a screen for chemopreventive efficacy. PMID- 11396158 TI - Immunohistochemical staining for DNA topoisomerase I, DNA topoisomerase II-alpha and p53 in gastric carcinomas. AB - New anticancer drugs targeting DNA topoisomerase I (topo I) are showing activity against gastric carcinomas. Laboratory studies have indicated that cells responsive to topo I targeted drugs have elevated levels of topo I, require active DNA replication and may require a functional apoptotic pathway. In this study, we evaluated these potential markers of topo I targeted drug sensitivity in 22 cases of primary gastric carcinoma. By immunohistochemical staining, we observed elevated topo I expression in 15 of 22 neoplasms (68%). By immunohistochemical staining for the proliferation marker DNA topoisomerase II alpha (topo II-alpha), we observed that 16 of 22 neoplasms (73%) had topo II alpha indices > than 50 indicating a large number of actively cycling tumor cells. Abnormal p53 expression was observed in 7 of the 22 cases (32%). Of the 22 cases of gastric carcinoma, 8 (36%) had high levels of topo I, a large number of cycling tumor cells and normal p53 expression. These are the molecular parameters that might suggest responsiveness to drugs targeting topo I. PMID- 11396159 TI - Inhibition of mouse melanoma cell proliferation by corticotropin-releasing hormone and its analogs. AB - BACKGROUND: Observations that epidermal cells release both corticotropin releasing hormone (CRH) and proopiome lanocortin (POMC) peptides has raised questions about the physiological relevance of this hypothalamo-pituitary-like system in mammalian skin. As CRH has shown anti-proliferative effects on cultured keratinocytes, we tested whether CRH can also regulate growth of melanoma cells. MATERIALS AND METHODS: CRH, [D-Glu20]-CRH, [D-Pro5]-CRH, acetyl-cyclo(30-33)[D Phe12,D-Glu20,Nle21,D-His32,Lys33,D-Nle38]-CRH(4-41), acetyl-cyclo(30-33)[D Phe12,Nle18,D-Glu20,Nle21,D-Ala32]-urotensin I(4-41), urocortin, and sauvagine were tested on Cloudman melanoma cell proliferation in culture and B16 melanoma tumor growth in C57B1/6 mice. Calcium-sensitive fluorescence measurements were used to examine the effect of CRH on intracellular Ca2+ signaling. The effects of CRH and [D-Glu20]-CRH on blood pressure were compared by measuring mean arterial pressure in anesthetized rats. RESULTS: CRH and six analogs were tested, and all demonstrated exceptional potency in inhibiting Cloudman cell proliferation in culture, with half-maximal effective concentrations ranging between 0.2 and 100 pM. The amplitude of ionomycin-induced Ca2+ influx into Cloudman cells grown in suspension was reduced by 50% after 48-hr exposure to CRH. Daily injections of CRH or [D-Glu20]-CRH, 100 micrograms/kg.day s.c., for 5 days, reduced net B16 tumor volume in mice by 30-60% compared to control animals. [D-Glu20]-CRH was less hypotensive compared to CRH, despite having similar anti-proliferative potency. CONCLUSION: CRH, and various analogs thereof, inhibit proliferation of Cloudman cells in culture, and inhibit B16 tumor growth rate in vivo, most likely by activation of endogenous CRH1 receptors and subsequent altered intracellular Ca2+ signaling. CRH analogs, such as [D-Glu20]-CRH, with less hypotensive activity may provide new directions of therapy for melanoma. PMID- 11396160 TI - Nobody needs "relative tumour sizes" to compare tumour growth curves. AB - Tumour growth curve experiments play an important role in experimental oncology and established methods exist for their unbiased interpretation. However, many authors till introduce systemic biases to their data by using the so-called "relative tumour size" transformation, inspite of wide awarance about the limitations of this latter method. With this communication and by using a fictive exaggeration of the problems connected with the application of this transformation we would like to appeal to both authors and reviewers likewise to ban this potential source of misinterpretation from the oncological literature. PMID- 11396161 TI - Serum levels of vasoactive intestinal peptide (VIP) in patients with adenocarcinomas of the gastrointestinal tract. AB - BACKGROUND: VIP acts as a neuroendocrine mediator under physiological conditions, with an important role in water and electrolyte secretion in the gut. Recent findings suggest that VIP also promotes growth and proliferation of normal as well as malignant cells. We have investigated the VIP-serum levels in patients with pancreatic cancer and colonic adenocarcinoma as compared to healthy controls. This was accompanied by immunohistochemical investigations and in vitro experiments to further define the role of the peptide in pancreatic and colorectal cancer. MATERIALS AND METHODS: Serum levels of VIP were evaluated under standardized conditions in a total of 135 patients; 45 patients had metastatic colorectal cancer, 45 suffered from metastatic pancreatic cancer, and 45 healthy volunteers served as controls. Human pancreatic and colorectal carcinoma cell lines were incubated over 5 days with VIP in increasing concentrations. RESULTS: In healthy controls, a median VIP-serum level of 42.44 +/- 2.540 pg/ml (range, 12.9-98.5 pg/ml) was found, while patients with pancreatic cancer had a median level of 40.58 +/- 3.013 pg/ml (range, 6.9-102.4 pg/ml). In patients with cancer originating in the colon, however, a median serum level of 116 +/- 10.14 pg/ml (range, 51.6-487 pg/ml) was found. While no difference between healthy controls and patients with pancreatic cancer could be detected (p = 0.6381), a significant difference between patients with colorectal cancer and healthy controls (p < 0.0001) and patients with pancreatic cancer (p < 0.0001) was demonstrated. The median VIP-concentrations found in the patients sera for pancreatic and colonic tumor patient groups, 40 pg/ml and 115 pg/ml respectively, had no significant effect on the proliferation of PANC-1 and HT29, inhibited ASPC-1, BxPC3, COLO201 and HCT-15 cells, and stimulated the growth of one pancreatic (CAPAN-1) and one colonic (COLO320DM) cell line under these conditions. CONCLUSIONS: As opposed to pancreatic cancer and healthy controls, patients in our series had elevated serum VIP-levels. Further studies are warranted to evaluate whether VIP can be used as a tumor marker in this disease. PMID- 11396162 TI - Inhibitory effect of steroidal alkaloids on drug transport and multidrug resistance in human cancer cells. AB - Intrinsic or acquired resistance of tumor cells to multiple cytotoxic drugs (multidrug resistance MDR) is a major cause of failure of cancer chemotherapy. MDR is often caused by elevated expression of drug transporters such as P glycoprotein (P-gp) or multidrug resistance protein (MRP). A number of compounds, termed chemosensitizers, have little or no cytotoxic action of their own, but inhibit (P-gp) or MRP-mediated drug export and are capable of sensitizing MDR cells to the cytotoxic effects of chemotherapeutic drugs. Here we examined the ability of steroidal alkaloids of plant origin, namely the Veratrum sp. alkaloid cyclopamine and the Lycopersicon sp. alkaloid tomatidine, to act as potent and effective chemosensitizers in multidrug resistant tumor cells. Drug uptake was determined by measuring accumulation of tetramethylrosamine in multidrug resistant NCI AdrR human adenocarcinoma cells. Resistance to adriamycin and vinblastine was determined by utilizing the MTT cell survival assay. Cyclopamine and tomatidine elevate tetramethylrosamine uptake by NCI AdrR cells and sensitize the cells to the cytotoxic action of adriamycin and vinblastine. In both cases these agents are comparable in patency and efficacy to verapamil, a reversal agent commonly used in MDR research. It is concluded that steroidal alkaloids of plant origin act as inhibitors of P-gp-mediated drug transport and multidrug resistance and therefore may serve as chemosensitizers in combination chemotherapy with conventional cytotoxic drugs for treating multidrug resistant cancer. PMID- 11396163 TI - An ab initio molecular orbital analysis of phosphorothioate mono-anion. AB - Ab initio calculations at the STO-3G level show that the energy of the highest occupied molecular orbital (HOMO) of the di-protonated phosphorothioate mono anion is 0.0421 a.u. higher than that of the HOMO of the corresponding phosphate mono-anion. The electron density in the phosphorothioate HOMO is highly focused on the sulfur atom. The results of this ab initio molecular orbital analysis are consistent with the model of the phosphorothioate sulfur as a soft nucleophile. The ab initio molecular orbital analysis presented in this paper will be useful for predicting and rationalizing the probability of reaction of phosphorothioate anions with electrophiles and helps to provide a basis for the rational design of phosphorothioate-based oligonucleotide drugs. PMID- 11396164 TI - A novel culture environment for generating mature human dentritic cells from peripheral blood CD14+ cells. AB - BACKGROUND: We recently demonstrated that supernatants from cultures of peripheral blood mononuclear cells (PBMC) activated with anti-CD3-specific antibody (ACD3S) can induce, upon brief exposure, tumor-reactive lymphocytes in cancer patients. Here, we report that ACD3S can also induce rapid and stable maturation of dendritic cells (DC) which can be used as antigen presenting cells in in vitro protocols and for cancer immunotherapy in vivo. MATERIALS AND METHODS: A short (4-day) priming of CD14+ monocytes with granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) followed by only a 24 hour-incubation in ACD3S, is sufficient to generate fully mature and stable DC. RESULTS: These DC (i) stimulated strong T cell proliferative responses in the mixed lymphocyte reaction, (ii) when pulsed with unfractionated peptides from autologous tumor membrane extracts activated CD4+ T cells which proliferated in response to the autologous tumor and CD8+ cytotoxic T cells (CTL) which specifically lyse autologous tumor targets and (iii) produced high levels of IL 12. CONCLUSION: ACD3S-treated DC are functionally superior to monocyte conditioned medium (MCM)-treated DC generated under the same short-term protocol and as efficient as DC induced by the standard 10-day protocol. Our data present an efficient and effective method for generating in a very short period of time, highly mature and functionally competent DC. PMID- 11396165 TI - TGF beta 1 induces caspase-dependent but death-receptor independent apoptosis in lymphoid cells. AB - Transforming growth factor beta 1 (TGF beta 1) is an antiproliferative and proapoptotic cytokine for normal B-cells, however many B-cell lymphomas have lost their response to TGF beta 1. The aim of this study was to identify the sequence of events in apoptosis induced by TGF beta 1 in an EBV negative, human B-cell lymphoma line (HT58). The proportion of apoptotic cells increased gradually (up to 72 hr) at an optimal dose range of 0.5-1.0 ng/ml. The induced cell death required the action of downstream caspases. Caspase activation was accompanied by an increase in the permeability of mitochondrial membranes, but there was no change in the expression of certain members of Bcl-2 family (Bcl-2, Bax, Bcl-XL). Similarly, none of the death receptors or ligands were involved in apoptosis induction. Further study will include the participation of TGF beta 1 target genes in the pore formation of mitochondrial membranes and/or the elimination of a putative survival signal. PMID- 11396166 TI - Detection of early gamma-postirradiation effects in murine spleen by proton NMR relaxation times. AB - BACKGROUND: It was our aim to evaluate the potential of proton relaxation times for the early detection of radiation-induced spleen changes. MATERIALS AND METHODS: Female Swiss mice were irradiated with doses ranging from 0.05 Gy to 4 Gy. The body weight, the spleen weight and the spleen water content of single animals were determined. Measurements of longitudinal (T1) and transversal (T2) proton relaxation times of the spleen samples were performed in a 0.47 T spectrometer. Histological examinations of the control and irradiated organs were performed. RESULTS: NMR measurements during the first five days after irradiation showed that total body gamma-irradiation with doses from 1.5 Gy to 4 Gy results in decreasing T1 of the murine spleen. Significant shortening in T2 was observed for the spleen of animals irradiated with a dose of 4 Gy. Histological examinations demonstrated subnormal architecture in slices derived from animals irradiated with 2 Gy and 4 Gy. CONCLUSION: The fluctuations of the spleen T1 and T2 of irradiated mice are correlated with relative spleen weight and can be used to estimate radiation induced changes in this organ. PMID- 11396167 TI - Increasing drug concentration in a rat lung tumor model by combining isolated lung perfusion with hypertensive chemotherapy. AB - Isolated left lung perfusion (ILP) with cisplatin was performed in Fisher 344 rats. Before perfusion, bolus injection with endothelin was given via the pulmonary artery. The vasoconstrictive potency was estimated by monitoring the perfusion pressure. The toxicity was estimated by tracking body weight change, survival rate after right pneumonectomy, arterial blood gas analysis and histological findings. To observe the pharmacokinetic changes, a solitary Methylcholanthrene-induced sarcoma model was established in a rat lung and the total platinum concentration in perfused lung and tumor tissues was measured. Perfusion pressure was increased significantly in a dose-dependent manner. Pulmonary toxicity from ILP with cisplatin was limited by the use of endothelin. Significantly higher levels of total platinum were obtained in tumors but not in normal lung tissues by endothelin injection before ILP than by ILP alone. The combination of ILP and hypertensive chemotherapy should be one of the available treatments for unresectable pulmonary carcinoma. PMID- 11396168 TI - The kinetics and mechanisms of the reaction of Mesna with cisplatin, oxiplatin and carboplatin. AB - BACKGROUND: Mesna is a sulfohydrate administered as a supportive drug in conjunction with oxazaphosphorines to prevent bladder toxicity from metabolites. When oxazaphosphorines are given simultaneously with platinum drugs, Mesna binds with platinum drugs as well. Previously we showed in cell culture, that Mesna reduces the efficacy of some platinum drugs. Here we elucidate the chemical reaction mechanism. MATERIAL AND METHODS: Cisplatin, Carboplatin and a novel platinum agent Oxiplatin were incubated with Mesna and the rate of disappearance of Mesna was measured, using an oxidation/reduction reaction between MTT and Mesna. RESULTS: All three platinum agents reacted with Mesna, but the chemical details differed largely. The molar ratios were 3:1, 2:1, and 1:1 for the reactions of Mesna with Oxiplatin, Cisplatin, and Carboplatin, respectively. The speed of the reaction followed a similar pattern, being fastest for Oxiplatin and slowest for Carboplatin. CONCLUSION: When considering the pharmacokinetics of Mesna and these platinum compounds and their reactivity, it appears unlikely that the reaction of Mesna with Carboplatin will become clinically relevant, while Cisplatin, might react with Mesna in patient serum. PMID- 11396169 TI - Secretion of a soluble T cell promoting factor by the human prostate adenocarcinoma cell line DU-145. AB - INTRODUCTION: A variety of cancers suppress host immune defenses by secretion of soluble factors. Conditioned media (CM) from numerous cancer cell lines possess the ability to suppress proliferation of activated T cells. The effects of CM from the prostate cancer cell line DU-145 on T cell activation was investigated. MATERIALS AND METHODS: Human PBMC, purified T cells and Jurkat T cells were treated with DU-145 CM. Proliferation, cell cycle, apoptosis, Fas expression/and cytokine secretion were assessed by thymidine incorporation, flow cytometry and ELISA. RESULTS: DU-145 CM increased proliferation of concanavalin-A (ConA) activated peripheral blood mononuclear cells (PBMC) increasing the percentage of cells in the S/G2 phase of cell cycle. Treatment of the Jurkat T cell line with DU-145 CM induced a potent proliferative response. ConA-induced proliferation of purified T cells from human PBMC and murine splenocytes was augmented in a dose dependent manner by addition of DU- 145 CM. DU- 145 CM treatment of ConA activated T cells induced an increase in interleukin-2 (IL-2) production. The soluble factor(s) responsible for promoting T cell proliferation was dependent on protein synthesis by the DU- 145 cells and possessed a molecular weight greater than 10 kDa. CONCLUSION: DU- 145 cells secrete a soluble factor(s) > 10 kDa, whose production is dependent on protein synthesis and which acts as a promoter of T cell activation. PMID- 11396170 TI - Augmented pro-apoptotic effects of TRAIL and proteasome inhibitor in human promonocytic leukemic U937 cells. AB - TRAIL, Tumor necrosis factor-related apoptosis-inducing ligand), a member of the TNF family, is known to be cytotoxic for a high proportion of tumor cell lines. However, successful application of TRAIL in tumor therapy may depend on finding other agents that can potentiate its antitumor effects. The present study showed that the cytostatic/cytotoxic TRAIL activity against U937 cells could be significantly augmented by proteasome inhibitor PSI, as revealed by MTT assay. Increased cytostatic/cytotoxic effect on U937 cells by TRAIL/PSI combined treatment was caused by apoptosis, as shown by an increased PARP cleavage rate. TRAIL/PSI did not affect the level of mRNA expression for TRAIL receptors (DR4, DR5, DcR1) and other apoptosis signal transduction molecules (TRADD, caspase-8). PMID- 11396171 TI - Therapeutic treatment of DMBA-induced mammary tumors with PPAR ligands. AB - The objective of this study was to evaluate the ability of troglitazone (a thiazolidinedione) and Wy-14,643 (a clofibrate) to inhibit progression of non detectable and detectable mammary tumors in rats induced by 7,12 dimethylbenz(a)anthracene (DMBA) when compared to those receiving no treatment or tamoxifen. Although not as effective as tamoxifen in decreasing overall tumor incidence, Wy-14,643 reduced the percentage and number of malignant tumors that developed when compared to both troglitazone and control. Treatment of detectable tumors with either Wy-14,643 or troglitazone induced regression or stasis of total tumor volume in 40-50% of the animals, compared to only 10% in control and 65% in tamoxifen treated animals. Moreover, each PPAR ligand was as effective as tamoxifen in preventing additional tumor development. In summary, both PPAR ligands were more effective than no treatment in preventing tumor progression once detected. However, only the PPAR-alpha activator, Wy-14,643 was able to reduce the development of malignant tumors when administered prior to detection. PMID- 11396172 TI - Suppression activity of pro-apoptotic gene products in cancer cells, a potential application for cancer gene therapy. AB - Overexpression of anti-apoptotic Bcl-2 and Bcl-XL proteins may play a role in the development of resistance to cancer therapy. We examined the expression of these proteins in prostate, breast, and ovarian cancer cells. We found that some of these cancer cell lines expressed high levels of Bcl-XL or Bcl-2., In order to develop an effective strategy to overcome the potential inhibition of cancer therapy by Bcl-2 and Bcl-XL, we tested the inhibitory ability of several pro apoptotic or tumor suppressor genes in these cells. The expression of these genes induced apoptosis or suppressed cell growth with variable efficiency in these cells. Harakiri (Hrk) appears to result in the greatest induction of apoptosis or inhibition of cell growth Mtd, bax and bcl-XS were also effective in inhibiting cell growth. Furthermore, transfection of Hrk, bax, or Mtd into these cells caused significantly less colony formation than in cells transfected with p53 or BRCA1. Therefore, these results suggest that Hrk, bax, and Mtd are potent therapeutic agents for cancers expressing high levels of Bcl-2 and Bcl-XL. PMID- 11396173 TI - In vivo evaluation of phosphorous-containing derivatives of dodecahydro-closo dodecaborate for boron neutron capture therapy of gliomas and sarcomas. AB - The in vivo uptake of dodecahydro-closo-dodecaborate derivatives substituted with phosphate- and bisphosphonate groups was evaluated in two different experimental tumor model systems and compared to other boronated and non-boronated compounds. These phosphorous-containing boron clusters may have potential for use in boron neutron capture therapy, a chemoradiotherapeutic form of cancer treatment. Using the F98 rat glioma as a brain tumor model in syngeneic Fischer rats, there was selective tumor uptake of the phosphate derivative with 21.5 micrograms boron/g tumor versus 5.2 micrograms/g normal brain and a tumor:blood ratio of 2.7. However, this compound was toxic to test animals and lethal at relatively low doses. The uptake of the bisphosphonate by the murine K8 osteosarcoma was approximately 18 micrograms boron/g tumor with a T:Bl ratio of 7.6 and a tumor:bone ratio of 1.5. This compound was non toxic to the test animals. The results indicate that phosphate- and bisphosphonate derivatives of dodecahydro closo-dodecaborate may have potential for BNCT of gliomas and osteosarcomas, respectively. PMID- 11396174 TI - Influence of beta-adrenergic antagonists, H1-receptor blockers, analgesics, diuretics, and quinolone antibiotics on the cellular accumulation of the anticancer drug, daunorubicin: P-glycoprotein modulation. AB - BACKGROUND: Treatment of patients with several drugs simultaneously may result in modulation of the naturally expressed P-glycoprotein (Pgp) at different tissues. With this possibility in mind, we have assessed the ability of different classes of drugs to modulate Pgp function in vitro. Modulation of the Pgp function was studied at in vitro drug concentrations comparable to therapeutic blood levels of the drugs. MATERIALS AND METHODS: Human blood brain barrier endothelial cells and human colon adenocarcinoma cells were transduced or transfected with the multidrug resistance gene (MDR1) to express Pgp. The uptake of fluorescent substrates of Pgp, Rhodamine 123 and daunorubicin, into these cells and NIH3T3/MDR1 and MDCK/MDR1 cells was measured by flow cytometry and in monolayers in the presence and absence of the different drugs. RESULTS: From the tested six H1-receptor blockers, seven beta-adrenergic antagonists, four analgesics, ten diuretics and five quinolons, five drugs inhibited Pgp at therapeutic blood levels and two at somewhat higher concentrations. Significant synergism for blocking Pgp could be demonstrated for several drugs. CONCLUSION: We conclude that administration of several drugs which modulate the function of Pgp to patients may adversely affect the natural function of this efflux pump and may cause drug-drug interactions induced side effects. PMID- 11396175 TI - Increasing specificity of anti-tumor therapy: cytotoxic protein delivery by non pathogenic clostridia under regulation of radio-induced promoters. AB - BACKGROUND: Pathogenic clostridia, genetically engineered to express therapeutic genes, will specifically target hypoxic regions in tumors. This specificity can be further improved if expression of these genes is controlled by a radio-induced promoter, leading to spatial and temporal control of gene expression. MATERIALS AND METHODS: Following administration of Clostridium spores to tumor bearing rats, normal tissue and tumoral specimens were compared for colonization. Clostridium was genetically modified to express tumor necrosis factor a or cytosine deaminase. Expression of these proteins was assayed. Northern blot hybridizations were used to detect genes which are radio-induced. RESULTS: Clostridium gave a selective colonization of tumors. The recombinant clostridia expressed in vitro and in vivo TNF alpha and cytosine deaminase. Clostridial SOS repair genes were induced at a dose of 2 Gy. CONCLUSIONS: Pathogenic Clostridium can be used for tumor specific delivery of therapeutic genes. The specificity can be improved via radio-induced promoters. Overall, this new gene delivery system can lead to an increase of the therapeutic ratio in cancer treatment. PMID- 11396176 TI - Methylselenocysteine modulates proliferation and apoptosis biomarkers in premalignant lesions of the rat mammary gland. AB - In the rat mammary carcinogenesis model, premalignant lesions known as intraductal proliferations (IDPs) are detectable within a few weeks after carcinogen treatment. These early transformed colonies are the precursors for the eventual formation of carcinomas. Our past research indicated that methylselenocysteine added to the diet of rats reduced the development of IDPs of all sizes (the size of each IDP was estimated operationally by the number of 5 micron serial sections showing the same pathology). The appearance of an IDP lesion represents a balance between cell proliferation and cell death. The modulation of these two cellular events by methylselenocysteine was investigated. The abdominal-inguinal mammary gland was excised 6 weeks after MNU administration. Proliferation and apoptosis were evaluated by BrdU labeling and the TUNEL assay, respectively. The expression levels of several cell cycle and apoptosis regulatory proteins, including cyclin D1, cyclin A, p27, p16, bcl-2, box and bak, were also assessed. All of the above endpoints were quantified by immunohistochemistry in paraffin-embedded sections. The results showed that the magnitude of the response to methylselenocysteine intervention seemed to depend on the size of the IDP lesion. For the purpose of this study, the small and large lesions were classified as those containing < or = 30 or > 30 serial sections, respectively. With the small lesions, methylselenocysteine significantly inhibited BrdU labeling and the expression of cyclin D1 and cyclin A, but increased the expression of p27. Interesting, only p27 was upregulated in the larger IDP lesions, while BrdU labeling and the cyclins were not affected. It is possible that the transformed phenotype becomes less sensitive to selenium mediated arrest of proliferation once it progresses to a more advanced pathological stage. In contrast, methylselenocysteine stimulated apoptosis (TUNEL assay) by 3 to 4 fold, and this increase was evident in both the small and large IDP lesions. Consistent with the induction of apoptosis, a reduced expression of bcl-2 was also observed in the methylselenocysteine group. In summary, our data suggest that exposure to methylselenocysteine blocks clonal expansion of premalignant lesions at an early stage. This is achieved by simultaneously modulating certain molecular pathways that are responsible for inhibiting cell proliferation and enhancing apoptosis. PMID- 11396177 TI - Regulation of gene expression and cell growth in vivo by tetracycline using the hollow fiber assay. AB - The hollow fiber assay presents a potentially unique tool to study the effects of regulated gene expression in cell lines that do not form tumors in vivo. The hollow fibers allow small molecules to pass freely through while keeping the cells within the fibers and segregated from host cells. OSp16.1 cells, derived from the U24 clone of the U2-OS osteogenic sarcoma tumor line, express the p16INK4a tumor suppressor under the regulation of tetracycline (tet) (Mitra J et al. Mol Cell Bio 19:3916, 1999). The in vitro induction of p16 in the OSp16.1 cell line is regulated by tet. The hollow fiber assay was used to determine whether the regulation of the p16 gene could be achieved in vivo, since these cells did not grow in the xenograft model. There were no differences in the in vivo growth pattern of U24 cells loaded into the hollow fibers with and without tet: 807% and 839% net growth, respectively. OSp16.1 cells in fibers in mice receiving 3.33 mg/kg/day tet had a 644% net growth after 21 days. There was a 194% net growth without tet. Immunoblotting of extracts prepared from the hollow fibers confirmed that p16 was induced in the absence of tet. These data demonstrate this assay is a useful tool for studying the effects of regulated gene expression in vivo. PMID- 11396179 TI - Modulation of multidrug resistance in a cancer cell line by anti-multidrug resistance-associated protein (MRP) ribozyme. AB - Multidrug resistance-associated protein (MRP) is the major candidate molecule responsible for non-P-glycoprotein (PGp)-mediated multidrug resistance. We used a hammerhead anti-MRP ribozyme (alpha MRP-Rz) to inactivate MRP function in a multidrug resistant cancer cell line, KB8-5. The beta-actin promoter-driven alpha MRP-Rz sequence (pH beta/alpha MRP-Rz) was introduced into KB8-5 cells (KB8 5/alpha MRP-Rz) and we evaluated growth of the cell line. The gene expression of multidrug resistance-related molecules was estimated. Drug sensitivity was estimated by MTT assay in vitro. MRP mRNA expression was decreased in KB8-5/alpha MRP-Rz cells. The MTT assay showed increased IC50 values or resistance to doxorubicin (DOX), etoposide (VP-16) and cisplatin (CDDP) in KB8-5/alpha MRP-Rz cells. No significant differences were observed in expression of multidrug resistance gene (MDR1), thymidylate synthase, glutathione S-transferase pi or topoisomerase II alpha. The hammerhead ribozyme-mediated simple suppression of MRP mRNA expression was not sufficient to reverse multidrug resistance in the cancer cell line KB8-5. PMID- 11396178 TI - Curcumin induced modulation of cell cycle and apoptosis in gastric and colon cancer cells. AB - We have investigated the chemopreventive role of curcumin in gastrointestinal cancers by studying the regulation of proliferation and apoptosis in gastric (KATO-III) and colon (HCT-116) cancer cells. Curcumin inhibited cell proliferation and induced G2/M arrest in HCT-116 cells. Investigation of the levels of cyclins E, D and B by immunoblot analysis showed cyclin B level was unaffected, whereas cyclin D and E levels declined with curcumin in both cell lines. Investigation of cyclin-dependent kinases, Cdk2 and Cdc2, showed activity of Cdc2, but not Cdk2, increased markedly in response to curcumin. In both cell lines, immunoblot analysis indicated that curcumin caused induction of apoptosis as evidenced by cleavage of PARP, caspase-3, and reduction in Bcl-XL levels. Curcumin also stimulated the activity of caspase-8, which initiates Fas signalling pathway of apoptosis. Curcumin therefore appears to exert its anticarcinogenic properties by inhibiting proliferation and inducing apoptosis in certain gastric and colon cancer cells. PMID- 11396180 TI - Effectiveness of a new device to retain carcinogenic compounds of tar from mainstream cigarette smoke for the prevention of smoking-associated tumors. AB - PURPOSE: Effectiveness of a new device inserted within the common cellulose acetate cigarette filter (named hypobaric chamber tar-removing system, HCTRS) to remove tar and its carcinogenic compounds from mainstream cigarette smoke (MCS). METHODS: Eighty HCTRS prototypes were mounted inside the cellulose acetate filter of commercial-brand cigarettes (13 mg tar) and smoked by eighty smoker volunteers. Tar retained by HCTRS prototypes was determined by weighing them before and after smoking. Subsequently, an aliquot (3-5 mg) of the tar retained by twenty randomly chosen HCTRS prototypes was analysed by high-performance liquid chromatography (HPLC) for the detection of polycyclic aromatic hydrocarbons (PAH). RESULTS: The mean value of tar retained was 12.80 mg/HCTRS prototype (S.D. = 5.31), thus showing that this simple device is capable of removing 98.5% of tar present in MCS. Minimal and maximal amounts of retained tar were 4.15 and 31.47 mg/HCTRS prototype, respectively. Moreover, these tar samples contained, although in differing amounts, each of the 16 priority pollutant PAH. A mean value of 259.42 ng/mg of tar (S.E.M. = 44.37) of the 16 main PAH was found in the tar of the 20 HCTRS prototypes examined. These data cogently demonstrate that the use of the HCTRS prototype can effectively eliminate about 100% of tar from MCS, thus reducing the inhalation of PAH (considered the most obvious carcinogenic tar components). CONCLUSIONS: The application of this device could be a suitable tool for effectively improving human health through the prevention of smoking-associated cancer. PMID- 11396181 TI - Complex regulation of CDK2 and G1 arrest during neuronal differentiation of human prostatic cancer TSU-Prl cells by staurosporine. AB - We are interested in the possibility of new prostate cancer therapy that would control tumor malignancy via the induction of terminal cell differentiation. We have previously reported that staurosporine induced remarkable inhibition of cell proliferation and neuronal differentiation in human prostatic cancer TSU-Pr1 cells. In the present study, we investigated the alteration of cyclin-dependent kinase (CDK) activities and cell cycle in differentiated TSU-Pr1 cells. Treatment of TSU-Pr1 cells with staurosporine resulted in G1 arrest and suppression of CDK2 activity. Protein levels of CDK2 were essentially unchanged during this time. p21 protein, however, rapidly increased for 6 hours after treatment with staurosporine. p27 protein also increased gradually for 12 to 72 hours after treatment. CDK2-bound p21 and CDK2-bound p27 also increased. These results suggest that an increase in p21 and p27 protein causes increased binding with CDK2 and inhibition of CDK2 activity. We propose that the complex regulation of CDK2 plays a key role in G1 arrest of TSU-Pr1 cells after treatment with staurosporine. PMID- 11396182 TI - Oral HCFU for prevention of hepatic metastasis in an experimental colorectal carcinoma. AB - BACKGROUND: An animal experimental study was performed to clarify the preventive effect of oral HCFU (1-hexylcarbamoyl-5-fluorouracil) on hepatic metastasis. MATERIALS AND METHODS: A total of 180 BALB/C mice received intraportal injection of Colon26 cancer cells followed by oral daily administration of HCFU to 5 groups divided according to the dose of HCFU, ranging from 0 (controls) to 7.5 mg/Kg for 21 days. These groups were independently assigned to 3 sets to investigate survival rates and degree of hepatic metastasis. RESULTS: Mean survivals in the treated groups were statistically longer than the controls except the 7.5 mg/Kg group. HCFU 7.5 and 5 mg/Kg groups showed the minimum liver weight on the 12th and 16th day, respectively these were significantly smaller than those of controls (p < 0.0001 in both). CONCLUSION: It is considered that oral HCFU could effectively suppress hepatic metastasis of the colon26 cancer cells and prolong the life of animals. This finding suggests the promising clinical application of oral HCFU for colorectal cancer after curative resection. PMID- 11396183 TI - Correlation between Topo II alpha expression and chemosensitivity testing for Topo II-targeting drugs in gynaecological carcinomas. AB - BACKGROUND: The purpose of this study was to investigate the correlation between topoisomerase II alpha (Topo II alpha) expression and the chemosensitivity to Topo II-targeting drugs in gynaecological carcinomas. MATERIALS AND METHODS: We analysed the expression of Topo II alpha, and then correlated this with the in vitro chemosensitivities of 43 gynaecological tumors using immunohistochemistry and a tetrazolium dye (MTT) assay. RESULTS: There was a significant correlation between the Topo II alpha index (the number of positive cells per 100 cells: %) and the tumor cell growth inhibition rate in culture (I.R.: %) for doxorubicin and etoposide (r = 0.631, p < 0.001 and r = 0.645, p < 0.001, respectively). The I.Rs for doxorubicin and etoposide in endometrial carcinomas were lower than those in ovarian carcinomas [38.2 +/- 22.8 vs 54.6 +/- 29.8, 37.3 +/- 19.8 vs 59.3 +/- 30.6] (p < 0.05, respectively). Furthermore, the number of high Topo II alpha index (over 30%) tumors in the ovarian carcinoma cases was higher than that in the endometrial carcinoma cases (63.1% vs 45.8%, p < 0.05). CONCLUSIONS: Our data suggest that the Topo II alpha index of a tumor is a reflection of its chemosensitivity to Topo II-targeting drugs. The use of this index may enable prediction of a clinical response to chemotherapy using Topo II-targeting drugs in gynaecological malignancies. PMID- 11396184 TI - Enhanced expression of cyclooxygenase-2 and nuclear beta-catenin are related to mutations in the APC gene in human colorectal cancer. AB - Mutational inactivation of the human tumour suppressor gene adenomatous polyposis coli (APC) results in constitutive activation of beta-catenin/T cell factor-4 (Tcf-4) mediated transcription of target genes. Up-regulation of cyclooxygenase-2 (COX-2) protein is frequently found in human colorectal cancer (CRC). We analysed 38 CRC for mutations in APC and beta-catenin and found an association between APC mutations and elevated COX-2 levels. Furthermore, APC mutations were predominantly observed in tumour tissues from the rectum compared to tumours of colonic origin. Western blot analysis revealed that nuclear beta-catenin levels were generally higher in tumours with APC mutations compared to tumours with wild type APC. However, there was also a higher level of nuclear beta-catenin in tumour compared to normal tissue, but nuclear Tcf-4 protein was constitutively expressed in tumour and normal tissue and showed no differences. An identified putative Tcf-4 binding element in the COX-2 promoter may partly explain the enhanced level of COX-2 and support the idea that COX-2 may be a downstream target of the APC/beta-catenin/Tcf-4 pathway. PMID- 11396185 TI - The antiproliferative effect of coumarins on several cancer cell lines. AB - Twenty-one coumarins were examined for their antiproliferative activity towards several cancer cell lines, namely lung carcinoma (A549), melanin pigment producing mouse melanoma (B16 melanoma 4A5), human T-cell leukemia (CCRF-HSB-2), and human gastric cancer, lymph node metastasized (TGBC11TKB). The structure activity relationship established from the results revealed that the 6,7 dihydroxy moiety had an important role for their antiproliferative activity. Analysis of cell cycle distribution indicated that esculetin-treated cells accumulated in the G1 (at 400 microM) or in S phase (at 100 microM). PMID- 11396186 TI - Immunohistochemical reactivity of anti-melanoma monoclonal antibody 225.28S in human breast cancer biopsies. AB - The high molecular weight melanoma-associated antigen, defined by murine monoclonal antibody (IgG1) 225.28S is largely expressed by melanoma cells and weakly expressed by other human tumors originating from neural crest. In this study, we analyzed the immunohistochemical reactivity of MoAb 225.28S in human breast cancer biopsies. A total of 92 breast cancer biopsies (66 infiltrating lobular and 26 infiltrating ductal carcinomas) were initially tested along with 26 melanomas (positive controls), 23 gastric/colonic adenocarcinomas and 13 neuroendocrine tumors. Forty-four out of 66 lobular breast carcinomas showed positive immunostaining with 225.28S MoAb as well as only 6 out of 26 infiltrating ductal histotype and 12 out of 26 melanomas. Conversely, gastric and colonic adenocarcinomas and neuroendocrine tumors were completely negative. The pattern of positivity in breast carcinomas was associated with malignant cells, rather than with the stroma or histiocytes infiltrating the lesions. Nonspecific cross-reactivity of 225.28S with breast carcinomas was excluded using a similar murine antithyreoglobulin MoAb, which gave negative staining in all biopsies. These results indicated that HMW-MAA or a similar sequence recognized by 225.28S MoAb is often expressed by lobular breast carcinomas but rarely by ductal adenocarcinomas. This seems to suggest that lobular breast carcinoma has common "ancestor" antigens with melanoma. PMID- 11396187 TI - Essential viral and cellular zinc and iron containing metalloproteins as targets for novel antiviral and anticancer agents: implications for prevention and therapy of viral diseases and cancer. AB - In this review the authors summarize the experimental data on the role of a selected group of metalloproteins, particularly viral (v) and cellular (c) zinc finger proteins (ZFP) and iron containing proteins which are involved in cell proliferation, neovascularization, apoptosis, and viral infection. Furthermore, this review summarizes the data embracing the hypothesis that disruption of certain metalloproteins by novel pharmacological agents is a key factor in controlling viral and proliferative diseases. The primary goal of this review is to show the potential therapeutic applications of ZFP disrupting agents, zinc chelators and iron chelators in the control of viral diseases and cancer. It is known that zinc or iron deficiency, resulting from exposure of culture cells to membrane-permeable Zn2+ or Fe(2+)-chelators, can induced apoptosis in virally transformed cells while normal cells remain unaffected under these conditions. Apoptosis is possibly due to simultaneous inactivation of vZFP, cZFP, and/or iron containing proteins, which are essential for maintenance of cellular and viral structure and which are activated in virally transformed cells. New insights concerning apoptosis, viral metalloproteins, and novel antiviral agents will also be reviewed. From the evidence reviewed, one can infer that development of a variety of drugs that control or neutralize vZFP may lead to a new therapeutic approach directed at controlling and preventing a wide spectrum of viral diseases and cancer. Furthermore, the results suggest that these agents may be useful to prevent transmission of viral diseases. Finally, this review not only points out the limits of our understanding of this system, but also directs scientists to opportunities for future research. PMID- 11396188 TI - Cytotoxic activity of saponins from Camassia leichtlinii against human oral tumor cell lines. AB - Five steroidal saponins from Camassia leichtlinii showed higher cytotoxicity against human oral squamous cell carcinoma cells HSC-2, as compared to normal human gingival fibroblasts HGF. The tumor specificity of saponins varied considerably from sample to sample, but was generally higher than that of tannins, flavonoids and prenylated compounds such as geranylgeraniol and vitamin K2 (MK-2). Agarose gel electrophoresis showed that the saponins failed to induce internucleosomal DNA fragmentation, but produced large DNA fragments in HSC-2 cells, whereas two saponin samples (compounds 1 and 5) induced internucleosomal DNA fragmentation in human promyelocytic leukemic HL-60 cells. In contrast to epigallocatechin gallate or gallic acid, the cytotoxic activity of saponins was not significantly affected by metals (Co2+, Cu2+, Fe3+) or by antioxidants (sodium ascorbate, N-acetyl-L-cysteine, catalase). Furthermore, the saponins did not produce radicals (detected by ESR spectroscopy) nor oxidation potential (measured by NO monitor). These data suggest that an oxidation-mediated mechanism is not involved in the cytotoxicity induced by the steroidal saponins. PMID- 11396189 TI - Combination effect of lignin F and natural products. AB - We investigated the effect of lignin F, isolated from the alkaline extract of the cone of Pinus parviflora Sieb. et Zucc, on the cytotoxic activity and radical intensity (measured by ESR spectroscopy) of various natural products. Lignin F slightly inhibited the proliferation of human oral tumor cell lines (human squamous cell carcinoma HSC-2, human salivary gland tumor HSG), but not that of human gingival fibroblast HGF, suggesting its tumor specific cytotoxic action. Lignin F enhanced the cytotoxic activity of vitamin K2, vitamin K3, sodium ascorbate (vitamin C), epigallocatechin gallate (EGCG) (a major component of green tea), gallic acid (structural unit of tannin), chlorogenic acid, and 6 tea extracts (Japanese green tea, Japanese barley tea, black tea, Chinese green tea, Chinese Jasmin tea, Chinese Oolong tea), to various extents. On the other hand, lignin inhibited the cytotoxic activity of curcumin and dopamine. ESR spectroscopy demonstrated that combination of lignin and vitamin K3, EGCG or gallic acid synergistically augmented the radical intensity. Lignin F enhanced the bactericidal activity of EGCG against E. coli. These data suggest the beneficial effect of the combination of lignin F and natural products. PMID- 11396190 TI - Development of an animal model for prostate cancer cell metastasis to adult human bone. AB - BACKGROUND: Prostate cancer metastases to bone are associated with significant morbidity and mortality. Presently, there is little known about the biological interaction between prostate cancer cells and bone. Development of an animal model using adult human bone will enhance our ability to study the biology of prostate cancer metastasis to bone. METHODS: Bone was harvested from patients undergoing total joint arthroplasty and implanted in the hindlimbs of pre-treated SCID mice. Two months after bone implantation 4 x 104 prostate cancer cells (PC-3 or LAPC-4) were injected near the bone implantation site. The animals were sacrificed approximately 8 to 12 weeks after the injections of the cells. Complete histological analysis including immunostaining was performed. RESULTS: Both the PC-3 and LAPC-4 prostate cancer cells homed to the human bone implant, specifically the reconstituted bone marrow cavity. Analysis of the bone-tumor interaction after injection of PC-3 cells revealed strong labeling for PTHrP, TNF alpha and IL-6, consistent with osteoclast recruitment and osteoclast activity. These cells also were positively stained for CK18. After cellular injection of LAPC-4 cells, there was strong labeling for TNF alpha, IL-6, and IL-1 (osteoclast recruitment and osteolytic activity). PTHrP staining was also noted. The bone cells were strongly stained for osteocalcin, and the tumor cells for PSA. CONCLUSIONS: These data suggest that the tumor cells may induce an osteolytic response to enhance their ability to metastasize to bone. This animal model allows us to study the biologic interaction between prostate cancer cells and human bone and may enhance our understanding of the events associated with prostate cancer metastasis to bone. PMID- 11396191 TI - Expression of hnRNP B1 in four major histological types of lung cancers. AB - Heterogeneous nuclear ribonucleoprotein B1 is one of the nuclear pre-mRNA binding proteins involved in RNA metabolism. Recently, over-expression of B1 has been reported to be useful in the early detection of squamous cell carcinomas of the lung. To elucidate its significance in other histological types of lung cancers, we carried out a comparative study, four major types of lung cancers and normal lung tissues. 37 surgical specimens were examined using a B1-specific monoclonal antibody (2B2). Immunohistochemically, 2B2 demonstrated B1 protein in the nuclei not only of squamous cell carcinoma (10/10) but also of adenocarcinoma (17/18), small cell carcinoma (5/5) and large cell carcinoma (3/4). A lesser amount of B1 protein was also detected in normal cells. Quantitative immunoblotting revealed that B1 expression was markedly higher in cancer tissues than normal tissues and it varied among the four histological types. To establish the usefulness of B1, a threshold should be set for over-expression. PMID- 11396192 TI - Alteration of X and Y chromosomes in human esophageal squamous cell carcinoma. AB - The incidence of human esophageal squamous cell carcinoma in males is well-known to be higher than in females and its biological action in male patients is generally much more aggressive than that of the female. Recently, aberrations and/or other abnormalities of the sex chromosomes, especially the Y chromosome, have been postulated to be involved in some of the differences in the incidence and/or biological action of human malignancies between male and female patients. Therefore, in this study, we examined abnormalities of the sex chromosomes in cell smears obtained from 30 male patients diagnosed with esophageal squamous cell carcinoma. In addition, TE series cell lines, derived from esophageal squamous cell carcinomas, were studied for sex chromosome abnormalities by utilizing a simultaneous double color fluorescent in situ hybridization (FISH) and these findings were correlated with various clinicopathological parameters in order to examine its likely biological significance. In esophageal squamous cell carcinoma, Y chromosome loss was detected in all cases studied (1.6-86.9%, mean 22.98 +/- 22.04%), but the loss of the X chromosome was encountered in only 6 of the cases (7.1-40.6%, mean 15.90 +/- 12.46%). There was no significant association between the rate of Y chromosome loss in carcinoma cells and any of the clinicopathological parameters examined including age and stage of the cancer. Loss of the Y chromosome was observed in only two cases of adjacent non pathological esophageal squamous cell epithelium. Among the TE series examined, the cell lines derived from male patients demonstrated loss of the Y chromosome in all cell lines (1.4-92.9%, mean 44.92 +/- 42.55%), but the great majority of cell lines derived from female patients were associated with the karyotype of XX. These results indicated that the loss of the Y chromosome is associated with the malignant phenotype in human esophageal squamous epithelium, but possibly not with biological behavior. These results also suggested that at least one X chromosome is indispensable for the survival of esophageal squamous cell carcinoma. PMID- 11396193 TI - Alterations of the 16q22.1 and 16q24.3 chromosomal loci in sporadic invasive breast carcinomas: correlation with proliferative activity, ploidy and hormonal status of the tumors. AB - BACKGROUND: Breast cancer is characterized by complex genetic alterations found in multiple chromosomal regions, most commonly losses of 17p, 16q, 8p and others. A number of tumor suppressor genes mapped on these loci have been investigated in mammary tumors, whereas other gene products are of unclear function and await identification. MATERIALS AND METHODS: We analyzed the loss of heterozygosity (LOH) of two chromosomal loci: a. 16q24.3 using the genetic markers D16S303, D16S3026 and D16S3407 and b. 16q22.1, the locus of E-cadherin gene, using the microsatelite markers D16S503, D16S752 and D16S512, in a series of 63 sporadic invasive breast carcinomas consisting of 56 ductal, 4 lobular and 3 tumors of mixed type. Our findings were correlated with proliferative activity, ploidy and hormonal status of the tumors. RESULTS: Fourteen (22.2%) tumors demonstrated LOH of 16q24.3. Allelic imbalance of the 16q22.1 locus was found in 19 of 61 informative cases (31%) and commonly coexisted with LOH of 16q24.3. A significant association was observed between LOH of D16S752 and the absence of progesterone receptors in tumor cells (p = 0.005). CONCLUSIONS: LOH of 16q24.3 and 16q22.1 are frequent genetic alterations in breast cancer and they do not seem to correlate with tumor cell proliferation or ploidy. The statistical association between LOH of 16q24.3 and progesterone receptors need to be further investigated in larger series. PMID- 11396194 TI - Role of serum tumor markers CA 125 and CEA in non-small cell lung cancer. AB - BACKGROUND: CA 125 and CEA are valuable serum tumor markers that can be used to monitor response to therapy in patients with various solid tumors. Systemic studies of CA125 and CEA have not been evaluated in lung cancer. In this study, we report the serum levels of CA 125 and compared it to CEA in newly diagnosed lung cancer and analyzed the serum levels of these markers pre- and post-therapy. MATERIALS AND METHODS: Two hundred and sixteen patients with newly diagnosed non small lung cancer were evaluated. CA 125 and CEA levels were correlated with stage and histopathology. RESULTS: CA 125 levels and CEA levels were shown to be lower in patients with early stage disease as compared to patients with unresectable or metastatic disease. CEA levels were significantly higher among patients with adenocarcinoma, while there was no statistically significant relationship between histology and CA 125. There was a statistically significant difference in the CEA and CA 125 levels dependent on tumor size. Thirty-seven patients were analyzed for responses to chemotherapy and responders are more likely to have decreases in CA 125 or CEA. CONCLUSION: When abnormally elevated inpatients witlrlung cancer, CA 125 and CEA are useful indicators of disease extent, a useful clinical therapeutic marker, and may potentially have important prognostic value. PMID- 11396195 TI - Usefulness of galactose oxidase-Schiff test in rectal mucus for screening of colorectal malignancy. AB - Based on a "field-effect" theory in colon carcinogenesis, and the expression of the disaccharide tumor marker D galactose-beta-[1-->3]-N-acetyl-D-galactosamine (Gal-GalNAc) in the rectal mucus of patients with cancer and precancer of the colon, Shamsuddin developed a simple, accurate, inexpensive, easy to perform and rapid (< or = 15 min) screening test for colonic cancer and precancerous lesions. In this study we examined 137 rectal mucus samples of randomly selected patients with colorectal malignancy or other colorectal diseases to confirm the sensitivity and specificity of this test in Croatia. Additionally, to test the validity of the "field-effect" theory, that the mucosa away from the obvious cancer will show abnormalities as a result of the generalized effect of the carcinogen throughout the entire field of the target tissue, we also monitored a subset of 53 patients post-operatively. Individuals free of colonic or any other malignancies served as control (n = 31). The rectal mucin was smeared on membrane filter and developed by a sequential reaction of galactose oxidase (GO) and Schiff's reagent. The test results were correlated to the findings from colonoscopy/surgery and histopathology. The sensitivity of the test was shown to be 100% and the specificity was 96.8% (p < 0.001). Interestingly, the test was positive in 60% (32 of 53) of the samples collected from patients after tumor resection, showing the persistence of the biochemical changes even though malignant tumors were removed, hence supporting the field-effect phenomenon of carcinogenic stimuli. Five patients out of these 32 (16%) postoperative cases with positive GO test had a tumor recurrence within a year (0.05 < p < 0.10), suggesting that a persistently positive GO test in this population may serve as a predictor of tumor recurrence. We conclude that Gal-GalNAc is an early and intermediate biomarker, suitable not only for the detection of malignancy in its inception, but also for monitoring of people at high risk for cancer, and the efficacy of the cancer therapy as well as secondary prevention by this technology. PMID- 11396196 TI - Effect of intratumoral OK-432 administration of thymidine phosphorylase expression in human gastric carcinoma. AB - It is known that thymidine phosphorylase (dThdPase) is increased in various types of malignant tumors and is induced by cytokines. In this study, we have investigated the effects of OK-432, which induces multiple cytokines, on dTHdPase expression and angiogenesis in human gastric carcinomas. We examined 25 patients who underwent gastrectomy for gastric carcinoma. OK-432 was directly injected in tumors in 16 (OK group) of 25 patients via endoscopy before operation and the other 9 patients were not treated (control group). The dThdPase activity in carcinoma tissues of the OK group was significantly higher than that of the control group (P < 0.05). The amounts of IL-1 alpha, IFN-alpha, and IFN-gamma in carcinomas in the OK group were significantly higher than in the controls (P < 0.05), and these were significantly correlated with the dThdPase activity. Intratumoral OK-432 administration enhances the expression of dThdPase in gastric carcinoma cells by inducing various cytokines. PMID- 11396197 TI - Comparative analysis of CA 242 and CA 19-9 serum tumor markers in colorectal cancer patients. A longitudinal evaluation. AB - OBJECTIVE: Following the encouraging results obtained on CA 242 as an adjunctive marker for colorectal cancer this study was designed to compare the clinical behavior of CA 242 to that of its related marker CA 19-9. PATIENTS AND METHODS: Sera from 630 patients with benign (n = 201) or malignant (n = 429) colorectal diseases were evaluated. Moreover, 50 patients with colorectal cancer were longitudinally monitored during. post-surgical follow-up for either a minimum of 5 years or until time of recurrence. Serum CEA, CA 19-9 and CA 242 levels were determined before treatment and at each scheduled follow-up. RESULTS: The distribution of CA 242 levels in colorectal cancer patients demonstrated a similar positivity rate (32.9%) compared to that of CA 19-9 (29.8%), although both sensitivities were lower than that of CEA (43.8%). Moreover, elevated CA 242 serum levels were found in metastatic disease (58.2%). A longitudinal evaluation demonstrated that serum CEA, CA 19-9 and CA 242 levels were elevated in 63.9%, 63.9% and 66.7% of recurrences. Combined evaluation of CEA, CA 19-9 and CA 242 serum levels in the overall population demonstrated a complementarity of CEA with the latter two markers. Conversely, a highly significant correlation was observed, suggesting that the two assays might recognize the same macromolecular complex. CONCLUSION: CA 242 determination does not seem to offer a particular advantage over CA 19-9, while CEA remains the marker of choice in monitoring colorectal cancer patients. PMID- 11396198 TI - Relationship between histologic grade and cytofluorometric cellular DNA and RNA content in primary bone tumors. AB - The diagnosis and grading of bone tumors remains a challenging problem. We studied the relationship between histologic grade and cytofluorometric cellular DNA and RNA content in 108 primary bone tumors. The data included DNA ploidy, mean DNA content (MDC), S-phase fraction (SPF), mean RNA content (MRC) and RNA/DNA ratio (RDR; MRC/MDC) which represents the RNA content normalized for the DNA content. Benign tumors had a diploid stem line with low MDC (mean; 1.04), low SPF (0.9), high MRC (2.41) and high RDR (2.31). Giant cell tumors of bone, which are locally aggressive benign tumors, showed diploidy with relatively higher MDC (1.07, p < 0.01) and SPF (2.6, p < 0.01) and lower MRC (1.81, p < 0.01) and RDR (1.69, p < 0.01). Similar results were obtained in low-grade sarcomas. In high grade sarcomas, the data depended on the histologic findings. Pleomorphic sarcomas such as osteosarcomas revealed aneuploidy with remarkably higher MDC (1.70 in osteosarcomas, p < 0.01) and SPF (6.5, p < 0.01), but lower RDR (1.70, p < 0.01). In contrast, small cell sarcomas, such as Ewing's sarcomas, showed diploidy with low MDC (1.11 in Ewing's sarcomas, N.S.) and SPF (2.5, p < 0.01) and extremely low RDR (1.34, p < 0.01). The RDR value was higher in well differentiated tumors than in primitive tumors, rendering it useful in grading bone tumors with a diploid stem line. By combining the RDR value with the MDC value, 96% of diploid sarcomas could be distinguished from benign tumors. These results indicate that cellular DNA and RNA content analysis may be of value in assessing the malignant potential of diploid as well as aneuploid bone sarcomas. PMID- 11396199 TI - The role of different biomarkers (DNA, PCNA, apoptosis and karyotype) in prognostic evaluation of superficial transitional cell bladder carcinoma. AB - BACKGROUND: In order to clarify the variable behaviour of transitional cell bladder carcinomas (TCBC) with same clinico-pathologic pattern, we investigated the prognostic significance of various biomarkers (PCNA, DNA, apoptosis, karyotype). MATERIALS AND METHODS: We studied 177 superficial TCBC (stage T1) undergoing transurethral resection (TUR). Analysis of biological indicators was performed on serial paraffin sections: DNA by static cytometry, karyotype by fluorescence in situ hybridisation (FISH), PCNA and apoptosis by immunohistochemistry. RESULTS: The most salient results are represented by prevalence of diploidy (56%), a mean PCNA labeling index (PCNA-LI) of 8.2%, nonrandom trisomies and tetrasomies of chromosome 7, and a high presence of apoptosis (in 72% of cases). These data were not related to histological grading. Multivariate analysis showed that only PCNA-LI is a an independent prognostic marker for patient survival (p = 0.01). Besides, we observed a worse prognosis in the presence of both very high PCNA indices and low or absent apoptosis. CONCLUSIONS: The results of this study suggest that tumor prognostic potential in TCBC should be evaluated on the basis of the association between PCNA cell kinetics and cell apoptosis information. PMID- 11396200 TI - Immunohistochemical MUC1 (DF3 antigen) expression of human esophageal squamous cell carcinoma. AB - MUC1 (DF3 antigen) is a member of a family of high molecular weight glycoproteins. Recent studies have demonstrated that MUC1 is expressed in tumors of various human organs and may function as an anti-adhesion molecule that inhibits cell-to-cell adhesion, inducing tumor metastasis. However, expression patterns of MUC1 have not yet been established in human esophageal carcinomas. In this study, we examined MUC1 expression and its histopathological localization in human esophageal squamous cell carcinoma. MUC1 immunoreactivity was found in 17 (32.1%) out of 53 esophageal squamous cell carcinomas, regardless of the depth of tumor invasion, vascular invasion or lymph node status. MUC1 expression was detected in the intramucosal part in 28.3% (15 out of 53) and in the invasive part in 32.6% (14 out of 43) of the esophageal carcinomas (no significant difference). These observations suggested that expression of MUC1 is an early event in cancer progression, but that it is not significantly associated with metastasis of human esophageal carcinomas. PMID- 11396201 TI - Expression of cyclooxygenase-2 in prostate adenocarcinoma and benign prostatic hyperplasia. AB - Elevated expression of cyclo-oxygenase (COX)-2 has been found in several human cancers, including prostate adenocarcinoma. To evaluate the potential prognostic role of COX-2 in prostate cancer, we assessed the expression of COX-2 in benign prostatic hyperplasia (BPH) and prostate cancer samples employing immunohistochemistry. COX-2 was over-expressed in 15 out of 18 (83%) prostate cancer samples whereas it was detected in only 22% (4 of 18) paired benign tissues. The intensity of immunostaining correlated with the tumor grading. In addition, COX-2 was expressed in 7 of the 22 (32%) BPH samples examined. The significance a COX-2 expression in the BPH samples is not known at present. This data suggest that COX-2 is over-expressed in prostate cancer and COX-2 inhibitors may be useful in combination chemotherapy or chemoprevention for prostate cancer. PMID- 11396202 TI - Influence of antioxidants and the CYP1A1 isoleucine to valine polymorphism on the smoking--lung cancer association. AB - To evaluate the association between CYP1A1 genotype and lung cancer risk and to assess the effect of CYP1A1 genotype and antioxidant supplementation on the smoking--lung cancer relationship we conducted a case-control study nested within a large cancer prevention trial cohort. Controls (n = 324) were matched to cases (n = 282) on age (+/- 5 years), intervention group and study clinic in a 1:1 ratio, using incidence density sampling. Genotype was determined by a PCR-based method and logistic regression was used to calculate relative risk estimates. Overall, we found no association between CYP1A1 genotype and lung cancer risk. CYP1A1 genotype did not modify the effect of smoking on lung cancer risk. However, in an examination of subgroups defined by randomized intervention assignment our findings suggest that alpha-tocopherol supplementation may reduce the risk of lung cancer associated with cumulative smoking exposure regardless of CYP1A1 genotype with the greatest effect seen among those with the variant CYP1A1 allele. PMID- 11396203 TI - Weekly intravenous recombinant humanized anti-P185HER2 monoclonal antibody (herceptin) plus docetaxel in patients with metastatic breast cancer: a pilot study. AB - BACKGROUND: In patients with HER2-positive metastatic breast cancer (MBC), combined treatment of herceptin (H) and chemotherapy (CT) improves time to progression, response rates and survival compared with CT alone. MATERIALS AND METHODS: We evaluated the safety and efficacy of weekly Docetaxel combined with weekly H as treatment in HER2 overexpressing MBC. RESULTS: Preliminary toxicity data from 12 patients and 76 cycles of D and 80 cycles of H were analysed. No G3/4 toxicity was observed. The most frequent non-hematologic toxicities were fatigue (2 patients G2, 2 patients G1), dyspepsia (1 patients G2, 3 patients G1), diarrhea (1 patient G2, 3 patients G1), and nausea (1 patient G2, 3 patients G1). Six partial responses have been observed in 12 patients (ORR 50%). CONCLUSIONS: The combination of weekly Docetaxel and Herceptin is well tolerated with significant anti-tumor activity. PMID- 11396204 TI - Detection of cervical lymph node metastases in nasopharyngeal carcinomas: comparison between technetium-99m methoxyisobutylisonitrile single photon emission computed tomography and computed tomography. AB - The aim of this study was to compare the effectiveness of technetium-99m methoxyisobutylisonitrile (Tc-99m MIBI) single photon emission computed tomography (SPECT) and computed tomography (CT) of head and neck in evaluating cervical lymph node (LN) metastasis in nasopharyngeal carcinomas (NPC). Fifty NPC patients with suspected cervical LN metastases underwent Tc-99m MIBI SPECT and CT of the head and neck to evaluate cervical LN metastases. Among the 50 patient.s cervical LN lesion were confirmed by biopsy histopathological results in 40 patients. For 22 LN lesions with discordant results between Tc-99m MIBI SPECT and CT Tc-99m MIBI SPECT could correctly detect 5 metastatic and 7 benign LN lesions, while could correctly detect 5 metastatic and 6 benign LN lesions. Tc-99m MIBI SPECT and CT could correctly detect all of the 18 metastatic LN lesions. Tc-99m MIBI SPECT showed a better specificity but a lower sensitivity for detecting cervical LN metastases in NPC when compared with CT. The combined use of Tc-99m MIBI SPECT and CT could increase the accuracy, compared with the single use of either Tc-99m MIBI SPECT or CT, to detect cervical LN metastases in NPC. PMID- 11396205 TI - S100 beta is a more reliable tumor marker in peripheral blood for patients with newly occurred melanoma metastases compared with MIA, albumin and lactate dehydrogenase. AB - Lactate-dehydroxynase (LDH) has been described as a leading blood parameter in patients with melanoma metastases. However, recent data indicates that levels of S100 as well as melanoma inhibiting activity (MIA) in peripheral blood, correlate with melanoma progression. The aim of this study was to evaluate tumor markers S100, MIA, LDH and albumin in peripheral blood of 373 melanoma patients. 284 patients presented with in-situ or UICC stage I/II, and 89 with stage III/IV (54 tumor-free, 29 with newly occurred metastases). For newly occurred metastases, sensitivity was highest for S100 in peripheral blood (0.86), followed by MIA (0.80), LDH (0.48), and albumin (0.15). Specificity for albumin (0.99) and LDH (0.98) was higher than for S100 (0.91) and MIA (0.62). This data indicate that S100 in peripheral blood as compared to MIA, LDH and albumin appears to be the most appropriate tumor marker for newly occurred melanoma metastases. PMID- 11396206 TI - Prostate-specific antigen density adjusted for the transition zone for staging clinically localized prostate cancer in Japanese patients with intermediate serum prostate-specific antigen levels. AB - The prostate-specific antigen (PSA) density of the transition zone (PSATZ) in 45 prostate cancer patients who received radical prostatectomy with a PSA value of 4.1-10 ng/ml was determined to see whether PSATZ was useful in the prediction of extracapsular invasion of prostate cancer. The value of PSATZ for the detection of extracapsular invasion was compared with that of PSA and PSA density (PSAD). Thirty-one patients (68.9%) had pathologically organ confined cancer while 14 (31.1%) had extracapsular disease. Patients with organ confined tumor had significantly lower PSAD and PSATZ than those with non-organ confined tumor. PSATZ was superior to PSA when analyzed by receiver operating characteristics curves. In those patients with a cut-off value of 1.0 ng/ml per ml of transition zone volume, the PSATZ had a sensitivity of 43% and a specificity of 90% for prediction of extracapsular extension. The present study demonstrated that PSATZ was superior to PSA as a predictor of extracapsular invasion in intermediate PSA levels. Measurement of PSATZ may be of additional value to indicate the need for radical prostatectomy. PMID- 11396207 TI - Copy number of cancer genes predict tumor grade and survival of pancreatic cancer patients. AB - Pancreatic cancer is on the increase. While means of early diagnosis are being sought, it continues to present late. Prognostication is based on patient and tumor characteristics, including expression or mutation of cancer-related genes. Few studies have examined the impact of the amplification of these genes on the outcome of pancreatic cancer. We have now used a non-radioisotopic slot-blot technique to relate gene copy numbers of p53, c-myc and K-ras to tumor grade and survival. Outcomes were corrected for patient characteristics, tumor location and TNM staging. The Kaplan-Meier test for likelihood of survival showed that increase in copy number of the two oncogenes and loss of p53 were associated with non-significant reduction in survival. When these variations in cancer gene copy numbers were, however, examined by logistic regression analysis in the context of patient and tumor characteristics, survival was negatively related to K-ras amplification (p = 0.0291). Tumor grade, but not survival was positively related to loss of p53 gene copy (p = 0.0131) as well as c-myc amplification (p = 0.0248). Thus using a simple non-radioisotopic technique for the detection of cancer gene copy number in association with patients and disease characteristics, we could predict survival on the one hand and tumor behavior on the other. Such information could be used to plan initial and follow-up therapy. PMID- 11396208 TI - Immunohistochemical expression of T, Tn and sialyl-Tn antigens and clinical outcome in human breast carcinoma. AB - BACKGROUND: The extent of expression of reactive T (Thomsen-Friedenreich), Tn and sialyl-Tn antigens has been assumed to predict carcinoma aggressiveness. We studied the expression of T, Tn and sialyl-Tn antigens in a relatively large cohort of breast carcinoma patients with known long-term outcome to assess the clinical and biological significance of these antigens. MATERIALS AND METHODS: T, Tn and sjalyl-Tn antigens were examined in 72 consecutive primary breast carcinomas by immunohistochemistry using well defined monoclonal antibodies and their semiquantitative values were correlated with established clinicopathologic prognostic parameters of the disease to determine their relationship with long term clinical outcome. RESULTS: Of the 72 carcinomas, 63 (87.5%) each expressed T or Tn antigens, while 16 (22%) expressed sialyl-Tn antigens. Most carcinomas (81%) expressed more than one of the antigens simultaneously, being the most frequent combination T/Tn antigen expression. No significant correlation was noted between the expression of T, Tn and sialyl-Tn antigens (whether individually or in combination) and the prognostic parameters including patient age, disease stage, tumor size, lymph node status, nuclear and histologic grades, histologic types, hormone receptor status and menopausal status. Univariate survival analyses showed that disease stage, tumour size and lymph node metastasis were significant predictors of overall survival. Interestingly, a significant inverse correlation was found between the Tn antigen expression (p = 0.04), as well as the combined T/Tn (p = 0.03) and Tn/sialyl-Tn (p = 0.02) antigen expressions and long-term overall survival. In a multivariate Cox proportional hazard model, disease stage and a negative or low Tn antigen expression emerged as significant predictors of overall survival. CONCLUSION: Our data suggested that the expression of T, Tn and sialyl-Tn antigens does not appear to predict the outcome of patients with breast carcinoma in a long-term run. Moreover, the findings signified a potential value for a negative or low Tn antigen expression in prognostic stratification of breast carcinomas. PMID- 11396209 TI - Expression of apoptosis and apoptosis-related proteins in microvessels of human ovarian epithelial tumors. AB - We performed an immunohistochemical analysis of apoptosis and the expression of apoptosis-related proteins (ARP) such as Fas and Fas ligand (FasL), bcl-2 and p53 in human ovarian epithelial tumors. Fas and FasL were abundant in endothelial cells of microvessels, and were observed, at times, in the myocytes of small arteries and veins, in parietal or in obstructive thrombi and fibroblasts. Apoptosis was also noticed in the endothelial cells of capillaries and sinuses. The expression of bcl-2 or p53 was rare. We found that the progression of tumor development was accompanied by considerable changes in the microvessels of ovarian tumors. These changes are probably related to the effect of ARP that are expressed by tumor epithelial cells, lymphocytes and macrophages. We suggest that the ARP are released as a result of necrosis of these cells and are taken up by cells of microvessels and by the cellular remnants of blood clots. The effect of tumors on the microvasculature can be regarded as an angiopathy that results in necrosis and hemorrhage within the tumoral tissue and enhances the progression of the malignancy. PMID- 11396210 TI - Annexin II overexpression is correlated with poor prognosis in human gastric carcinoma. AB - Annexins belong to a family of the calcium-dependent phospholipid binding proteins. They are also substrates of receptor tyrosine kinases. Overexpression of Annexin II, which has been reported in various carcinomas, is thought to be associated with cell proliferation, differentiation and cell-cell adhesion in the pathogenesis of carcinoma, but the functions of Annexins have not been fully elucidated. In this study, we investigated the role of Annexin II (p36) and its relationship with c-erbB-2 overexpression in gastric carcinoma. We studied Annexin II expression using Western blot analysis in 8 human gastric carcinoma cell lines and expression of Annexin II and c-erbB-2 using, immunohistochemistry in 153 primary gastric carcinomas. Western blot revealed that Annexin II was expressed in 8 human gastric carcinoma cell lines. It was more strongly expressed in the cell membrane than in the cytoplasm of tumor cells in primary gastric carcinoma tissues. Thirty-three percent of all cases were immunopositive for Annexin II, overexpression of which was more frequent in differentiated type (p = 0.0009), lymph node, metastasis (p = 0.0147) and venous invasion (p = 0.0092). Annexin II and c-erbB-2 overexpression were significantly correlated p = 0.0002) and patients with Annexin II had poorer prognoses (p = 0.0066). Multivariate analysis showed that immunopositivity of both Annexin II and c-erbB-2 was an independent and poor prognostic factor (p = 0.0037). In conclusion, Annexin II was overexpressed in advanced gastric carcinomas and it could contribute to the progression of gastric carcinoma. PMID- 11396211 TI - Gastric epithelium proliferation in early Hp+ and Hp- gastritis: a flow cytometry study. AB - BACKGROUND: Helicobacter pylori increases cellular turn-over causing hyperproliferation and possible assumption of neoplastic characteristics by the gastric epithelial cells. To verify whether patients at risk of cancer can be identified at the very first stages of gastric disease, we studied a sample of patients affected by early Hp+ and Hp- gastritis by flow cytometry and assessed the methods commonly adopted to study gastric cell proliferation. METHODS: 48 fresh biopsies taken from the gastric antrum and body of 24 patients who had undergone endoscopy for dyspepsia, and 48 paraffin-embedded antrum and body biopsies taken from the files of our Department were studied by flow-cytometry. The following markers and parameters were considered: S-phase, proliferation index, PCNA and ploidy. RESULTS: No correlation was found between Hp+ or Hp- gastritis and gastric cell proliferation and no cases of aneuploidy were observed. Gastric proliferation was found to vary depending on the methods, markers and type of biopsy employed. Furthermore, proliferation expressed by PCNA was significantly different in antrum and body. CONCLUSIONS: The commonly studied proliferation markers do not allow the early detection of patients at risk of gastric cancer by flow cytometry. Proliferation differences between body and antrum must be taken into account in the investigation of gastric diseases. PMID- 11396212 TI - Serum levels of tumor necrosis factor-alpha and nutritional status in pancreatic cancer patients. AB - BACKGROUND: Tumour necrosis factor-alpha (TNF-alpha) is a multifunctional cytokine associated with cancer-related cachexia. In this study we evaluated serum levels of TNF-alpha in pancreatic cancer patients and investigated their relationships to cachexia. PATIENTS AND METHODS: Serum TNF-alpha levels were determined in 63 patients with pancreatic cancer using an enzyme immunoassay specific for human TNF-alpha. RESULTS: Serum TNF-alpha levels were detected in 36.5% of patients. Patients with metastatic disease showed significantly higher positive serum TNF-alpha rate compared to those with non-metastatic disease. Patients with detectable serum TNF-alpha levels had significantly lower body weight and body mass index, lower haematocrit and haemoglobin values, and lower serum total protein and albumin levels compared to those with undetectable TNF alpha levels. Serum TNF-alpha levels were inversely correlated with body weight, body mass index, haematocrit, haemoglobin, and serum protein and albumin levels. CONCLUSIONS: TNF-alpha levels are detectable in the serum of pancreatic cancer patients, particularly in those with advanced disease, and these levels correlate with poor nutritional status. PMID- 11396213 TI - Limited effectiveness of extended lymph-node dissection for node-negative patients with proximal gastric cancer. AB - BACKGROUND: The optimal surgical treatment with respect to the extent of lymph node dissection for node-negative patients with gastric cancer remains to be established. MATERIALS AND METHODS: A total of 101 node-negative patients with proximal gastric cancer (62 males and 39 females; age range 33 to 79 years; mean 58.0 years), who had undergone curative total gastrectomy, were retrospectively evaluated to determine whether any correlation existed between survival and the extent of lymph-node dissection (D1, limited; D2, extended lymph-node dissection). RESULTS: The 10-year survival rates of patients with T1 (n = 59), T2 (n = 31) or T3 tumors (n = 11) were 100%, 90.0% and 46.7%, respectively. Significant differences in survival were found between patients with T1 and T2 tumors (p = 0.018), T2 and T3 tumors (p = 0.003), and T1 and T3 tumors (p < 0.0001). Despite the fact that only 9 patients with a T1 tumor underwent a D2 lymph-node dissection, all other patients had an excellent prognosis. On the other hand, the 10-year survival rates of patients with T2 or T3 tumors who underwent a D1 or D2 lymph-node dissection were 83.3% and 76.8%, respectively, representing no significant difference between the two procedures for advanced stage cases (p = 0.590). Multivariate analysis showed that depth of invasion was the only statistically significant prognostic factor (p < 0.0001; relative risk, 19.018). CONCLUSIONS: Conventional radical prophylactic D2 lymph-node dissection does not improve the survival of node-negative patients with proximal gastric cancer when compared to limited D1 dissection. PMID- 11396214 TI - Cross-correlation of serum chromogranin A, %-F-PSA and bone scans in prostate cancer diagnosis. AB - Circulating Chromogranin A (CgA), total PSA (TPSA) and F-PSA concentrations were measured in 211 patients (pt) with newly diagnosed prostate cancer (PC) and in 25 controls with benign prostatic hypertrophy (BPH). TPSA values ranging 3.5-5.5 ng/ml were found in 14 PC pt (6.6%), 5.5-9.9 ng/ml in 29 pt (13.7%), 10-19.9 ng/ml in 75 pt (35.6%), 20-50 ng/ml in 64 pt (30.3%) and > 50 ng/ml in 29 pt (13.7%). In those groups of PC pt false negative % F-PSA level > 18 was respectively measured in 0 out of 14, 2 out of 29 (6.9%) 6 out of 75 (8.0%), 61 out of 4 (9.4%) and 6 out of 29 (20.7%) pt, or totally in 20 out of 211 (9.5%) pt. Among 20 PC pt with false negative %F-PSA data elevated CgA level (> 80 ng/ml) was found in 18 subjects (18 out of 20 90%) or respectively in 0, 1/2 (50%), 516 (83%), 6 out of 6 (100%) and 6 out of 6 (100%) patients. Bone scintigraphy was performed in all pt with TPSA concentration > 10 ng/ml at the time of diagnosis. Bone lesions were respectively found in 4 out of 75 (5.3%) pt with TPSA 10-20 ng/ml, 12 out of 64 (14%) pt with TPSA level from 20-50 mg/ml and in 25k9 (75.9%) pt with. TPSA above 50 ng/ml. Overall osseous metastases were recorded in 41 out of 211 pt (19.4%) with newly diagnosed PC and in 18 of these Stage D2 pt (43.9%) elevated CgA concentration were measured Among them elevated CgA level and tumor dissemination matched with false negative %F-PSA parameter (> 18%) in 4 out of 18 (22.2%) pt as well as in 37 out of 191 (19.4%) pt with %F-PSA < 18% (p > > 0.05). In parallel, a positive CgA level in newly presented PC pt was closely associated with %F-PSA false negativity (18 out of 20, 90%). A negative correlation between TPSA elevation and the magnitude of CgA serotest level indicate differences in their biological origin and activities. According to the data reported herein we advocate the assessment of serum Chromogranin A concentration in first presented patients with clinically proven PC, elevated T PSA level and %F-PSA parameter > 18%. Neuroendocrine structures are resistant toward hormonal treatment and hence CgA measurement is strongly suggested in all candidates for a systemic hormone therapy. PMID- 11396215 TI - Docetaxel and epirubicin plus G-CSF as mobilizing treatment to support high-dose chemotherapy in breast cancer. AB - BACKGROUND: In order to combine an active regimen with a simultaneous efficient mobilization of peripheral blood precursor cells (PBPC), we explored the combination of Docetaxel 75 mg/m2 and Epirubicin 120 mg/m2 with G-CSF 5 mcg/Kg/day s.c. to mobilize PBPC in breast cancer patients to support high-dose chemotherapy (HDC). PATIENTS AND METHODS: Forty patients were enrolled: 27 high risk and 13 metastatic. The entire procedure, including chemotherapy and PBPC collection, was on an outpatient basis. RESULTS: The median day of starting apheresis was day +10 (range 10-12) and the average value of circulating CD34+ cells at peak was 175/microliter (range 33-403). The median yield of CD34+ cells per apheresis was 8.76 x 10(6)/Kg (range 1.83-27.87). None of the patients developed side effects which required hospitalization. All patients enrolled successively received HDC as consolidation treatment. High risk patients received one and metastatic patients two HDC with PBPC reinfusion. All patients obtained a complete engraftment. No significant differences between high-risk and metastatic patients were observed. CONCLUSIONS: Our study suggests that the combination of Docetaxel, Epirubicin, and G-CSF is feasible, safe and efficient outpatient mobilizing treatment for patients with breast cancer receiving HDC. PMID- 11396216 TI - Automatic search for optimal conditions in clinical studies. AB - BACKGROUND: A program for an automatic search for the optimal conditions of important prognostic factors in clinical studies was developed. METHODS: The program was developed for the following steps: (1) Input of the maximum and minimum values and of the interval of the variable to be investigated. Automatic calculation of the Cut Points. (2) Division of the patient data into two groups at every Cut Point and calculation of survival rates. (3) Sequential calculation of P-values and chi-square values for the two sets of survival rates. To determine the usefulness of this program, the optimal irradiation dose was searched for 537 patients with non-small cell lung cancer. RESULTS: The P-value reached its minimum value and the chi-square value its maximum when the Cut Point was 5,925 cGy (0.0001 and 30.18). Between 5,925 cGy and 6,900 cGy, the P-value stayed at less than 0.05. CONCLUSION: Artificial errors in grouping by prognostic factors can be avoided and the search for optimal conditions can be conducted automatically and scientifically. PMID- 11396217 TI - Value of 18F-fluoro-2-deoxyglucose positron emission tomography in the evaluation of recurrent colorectal cancer. AB - To evaluate the clinical value of positron emission tomography (PET) with 18F fluoro-2-deoxyglucose (FDG) in recurrent colorectal cancer, the records of 33 patients who underwent 34 FDG-PET scans were reviewed and compared with computed tomography (CT) and carcinoembryonic antigen (CEA). The final diagnosis was based on operative, histopathological findings or clinical follow-up > 1 year. FDG-PET detected all 15 patients with recurrent diseases (9 local recurrences, 4 lymphatic metastases, 2 hepatic metastases and 8 pulmonary metastases) and 1 primary lung cancer. However, there were 3 false positive cases. The overall sensitivity and specificity of FDG-PET were 100% and 83%, respectively. The sensitivity and specificity of CEA were 33% and 86%, respectively, for detecting recurrent colorectal cancer. Abdominal CT had a sensitivity and specificity of 78% and 61% for detecting local recurrence and detected one lymphatic and one hepatic metastasis. In conclusion, the FDG-PET was more accurate than CT and CEA for the detection of recurrent colorectal cancer. In addition, it accurately detected distant metastases or occult secondary malignancy and may affect clinical management. PMID- 11396218 TI - Prognostic factors in elderly patients with unresectable non-small cell lung cancer. AB - BACKGROUND: The number of elderly patients with lung cancer is rapidly increasing and their management is an important issue. PATIENTS AND METHODS: 109 patients aged over 75 years with unresectable non-small cell lung cancer were assessed to define the prognostic factors. The median age was 80 years in a range of 76 to 95. The overall median survival time was 6.3 months. Fifty-one patients underwent chemotherapy and/or thoracic radiotherapy whilst the others received best supportive care. RESULTS: Multivariate Cox regression model showed performance status (PS) (p = 0.0063) and stage of disease (p = 0.0158) to be independent prognostic factors for survival. In seventy-six patients with a good PS of 0-1, choice of treatment (p = 0.0225) and hyponatremia (p = 0.0302) were the predictors for survival. CONCLUSIONS: PS, treatment and serum sodium level were important factors for survival, and most patients with good PS were able to undergo the treatment and have a good outcome. PMID- 11396219 TI - A pilot study of AFL-T (doxorubicin, 5-fluorouracil, leucovorin, and tamoxifen) combination chemotherapy for hormone-refractory prostate cancer. AB - BACKGROUND: Tamoxifen had been used to treat advanced prostate cancer with limited success. In vitro data suggested that tamoxifen could enhance the cytotoxic effect of chemotherapeutic agents, including doxorubicin, on prostate cancer cell lines. We applied this observation into a phase II trial for patients with hormone refractory prostate cancer (HRPC). PATIENTS AND METHODS: The AFL-T regimen consisted of doxorubicin 30 mg/m2/day on day 1; 5-FU 2,000 mg/m2/day 24 hour infusion and leucovorin 200 mg/m2/day 24-hour infusion on days 15 and 29; tamoxifen 50 mg/m2 four times daily on days 1, 2, 16, 17, 30, and 31. The protocol was designed to be of low dose-intensity and tolerable to most HRPC patients who may have reduced bone marrow reserve and poor renal function. Between Feb. 1994 and April 1999, 17 patients (median age 67, range 60-81) with HRPC were enrolled. Extensive hormonal manipulations had been done prior to the chemotherapy. Three patients had measurable diseases, 14 had only bone metastases, and all had elevated PSA levels (median 498 ng/ml, range 7.4-3,970 ng/ml). RESULTS: All 17 patients were eligible for analysis of toxicity. ECOG Grade III/IV leukopenia and thrombocytopenia occurred in 1 and 3 patients, respectively. There was no febrile neutropenia; there was no treatment-related mortality. Grade III/IV nausea, vomiting, mucositis, and diarrhea were noted in 0, 0, 1 and 0 patient, respectively. There was no venous thrombosis. One partial response, 1 stable disease, and 1 progressive disease were found in the three patients with measurable lesions. Eleven of the 17 patients (64.7%, 95% confidence interval: 41-88%) who were eligible for the evaluation of PSA response (PSA decrease > 50% for at least 6 weeks) were responders. The median overall and progression-free survivals were 13 and 7 months, respectively. Seventy-six percent of patients showed decreased analgesic usage or enhanced performance status. CONCLUSION: AFL-T, that has a low toxicity profile, is comparable to most other active regimens in terms of the PSA response rate. Randomized trials are needed to determine if there exists a survival benefit for patients with HRPC. PMID- 11396220 TI - Bimodal expression of heparin-binding EGF-like growth factor in colonic neoplasms. AB - BACKGROUND: Recent studies have demonstrated that heparin-binding epidermal growth factor-like growth factor (HB-EGF) plays a role in carcinogenesis and carcinoma progression. In this study we investigated the expression of HB-EGF in human colonic non-neoplastic and neoplastic tissues. MATERIALS AND METHODS: We performed immunohistochemistry using a polyclonal antibody against HB-EGF for normal colon hyperplastic polyps, adenomas and adenocarcinomas. RESULTS: Normal human colon and hyperplastic polyps did not express HB-EGF. In adenomas with moderate or severe dysplasia, HB-EGF was positive in 92% of the cases, whereas only 14.6% of those with mild dysplasia, expressed HB-EGF (p < 0.0001). HB-EGF expression was observed in 75% of carcinoma-in-adenoma cases. In adenocarcinomas, the incidence of HB-EGF expression significantly decreased as compared to adenomas with moderate or severe dysplasia (p < 0.0001) and CIA (p = 0.0005), with only 27.5% of the cases being classified as positive. In adenocarcinomas, HB EGF expression was inversely linked to carcinoma differentiation (p = 0.0003) and lymph node metastasis (p = 0.0358). CONCLUSION: Our results demonstrated the bimodal expression of HB-EGF in colonic neoplasms and suggested that HB-EGF may play a role in colonic carcinogenesis and at an early phase of the progression of colonic adenocarcinoma. PMID- 11396221 TI - Implication of laminin and collagen type IV expression in the progression of breast carcinoma. AB - BACKGROUND: The prognostic variability in breast cancer patients prompted the authors to investigate specific biological markers for the identification of high risk breast cancer groups. In the present study, attention was focused on the interaction between tumor cells and the extracellular matrix, an important requisite in the metastatic process. MATERIALS AND METHODS: Fifty-six primary breast carcinoma specimens obtained by mastectomy or quadrantectomy plus axillary dissection were examined with immunohistochemistry, for the determination of laminin, collagen type IV and hormone receptor expression and with static cytometry, for the determination of the DNA content. RESULTS: Laminin and collagen type IV expression was observed on the membrane and in the cytoplasm of neoplastic cells. Laminin and collagen type IV were present, respectively, in 85.4% and 73.8% of the cases which showed recurrence. CONCLUSION: The results of this study have shown that high expression of laminin and collagen type IV may have a value in the prognosis of disease free survival and may be linked to other classical clinical, histological and biological parameters in the evaluation of breast cancer patients. PMID- 11396222 TI - Cysts of the adrenal gland: revision of a 15-year experience. AB - Ten specimens of adrenal cyst resected during the period 1984-1999 were re examined. Thorough examination of multiple sections and the use of immunohistochemistry allowed to change the recorded diagnosis in 8 cases: three epithelial cysts and five pseudocysts were redefined as endothelial cysts. All 10 cysts were of the endothelial type. A synthetic review of the current knowledge about the pathogenesis, the classification and the clinical aspects of this rare disease is presented. PMID- 11396223 TI - Extra-tumor perineural invasion predicts postoperative development of peritoneal dissemination in pancreatic ductal adenocarcinoma. AB - Peritoneal dissemination is one of the major patterns of postoperative recurrence and a major cause of death in pancreatic ductal adenocarcinoma. To predict the development of postoperative peritoneal dissemination (PPD), correlations between the occurrence of PPD and clinicopathological factors were analyzed in 23 patients with pancreatic ductal carcinoma who underwent tumor resection and were carefully monitored after the operation. Fifteen cases (65.2%) developed PPD, but the other 8 cases (34.8%) did not meet the criteria for a diagnosis of PPD. Location of the tumor in the pancreatic tail or body (p = 0.0041, HR = 10.827, 95% CI = 2.126-55.127) and the presence of extra-tumoral perineural invasion (ETNI) (p = 0.0490, HR = 0.114, 95% CI = 0.013-0.990) were found to have predictive value for the development of PPD. These results suggested that intensive therapy selectively focused on the peritoneum in cases with ETNI, might prevent the development of PPD and reduce the side effects of treatment in cases without ETNI. PMID- 11396224 TI - Prognostic features and treatment outcome in patients with nasopharyngeal carcinoma: an experience of 20 years. AB - BACKGROUND: The best treatment of Nasopharyngeal Carcinoma (NPC) is still an open question. The purpose of this retrospective study was to determine risk factors that affect locoregional control and treatment outcome of NPC patients after radiotherapy, with or without chemotherapy. METHODS: Between January 1976 and December 1996, 66 consecutive patients (stage I = 0; stage II = 13; stage III = 32; stage IV = 21) were given definitive radiotherapy at a single Institution. Concurrent or adjuvant chemotherapy was also given to 14 of them (21%). Multivariate analysis was performed to evaluate age, T stage, N stage, radiotherapy dose, histology, chemotherapy bone of skull erosions or cranial nerve palsies and base of skull involvement as prognostic factors of locoregional control and overall survival. RESULTS: By the end of January 2000, after a median follow-up of 66 months and a minimal follow-up of 36 months, the event-free overall survival rate of 5 years was 48% and the overall survival 54%. Risk factor analysis revealed that radiotherapy dose, age and stage were the most important factors for overall survival of these patients. The 5 year overall survival was 89% for stage II and 49% for stage III-IV (p = 0.004), 62% for dose higher than 60 Gy and 20% for dose below 60 Gy (p = 0.007), 62% for age below 65 years and 36% for age higher than 65 years (p = 0.027). The concurrent or adjuvant chemotherapy did not have prognostic significance. CONCLUSIONS: We confirm the need to determine the risk factors in patients with NPC. The choice of treatment, whether radiotherapy alone, at dose > 60 Gy, or radiotherapy plus chemotherapy, should be made after identification of patients with high risk disease, suitable for the combined modality. PMID- 11396225 TI - Unusual spread of liposarcoma. AB - BACKGROUND: The aim of our study was to investigate the unusual pattern of metastatic spread of liposarcoma. MATERIALS AND METHODS: We retrospectively studied thirty-two patients with liposarcoma (in seventeen located in the retroperitoneum and in fifteen in the extremities) who were managed at our institution over a ten-year period. Six patients with extremity liposarcoma developed local recurrence and or distal metastases and in three the site of the distal spread was other than the lungs. In only one patient with retroperitoneal liposarcoma we observed extra-pulmonary spread. The histologic type of all four cases with unusual spread was myxoid, with small areas of round cell differentiation in two cases. RESULTS: The survival of the patients after the initial recurrence was relatively prolonged (6, 1.5, 8 and 3 years with the last patient alive and well). CONCLUSION: Physicians should maintain a low threshold for initiating evaluation of relatively minor symptoms in patients with liposarcoma, as these symptoms may reflect the first manifestation of an extrapulmonary metastasis. Such an approach should result in earlier detection and hopefully improve survival for patients with liposarcoma. PMID- 11396226 TI - Angiogenic factors: vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (b-FGF) are not necessarily elevated in patients with advanced renal cell carcinoma. AB - Serum analysis of Vascular Endothelial Growth Factor (VEGF) and basic Fibroblast Growth Factor (b-FGF) levels were studied in 53 patients with renal cell carcinoma (RCC). Approximately 2/3 of the patients had disseminated disease at diagnosis, the remainder had no evidence of metastases. The results confirmed that VEGF has a major role in the angiogenesis of RCC. No correlation was observed between VEGF and/or b-FGF and the presence or absence of metastases, nor was any correlation observed between VEGF and/or b-FGF and patient survival. Thus, to utilise VEGF and/or b-FGF as a tumour marker at the time of diagnosis to predict patients with a high risk of progression, where an adjuvant therapeutic approach would be of great value, seems to be limited. Not all patients with RCC exhibited elevated serum levels of VEGF and/or b-FGF. No correlation was observed between tumour stage and serum levels of these angiogenic peptides. PMID- 11396227 TI - Phase II study of a combination of cisplatin, all-trans-retinoic acid and interferon-alpha in squamous cell carcinoma: clinical results and pharmacokinetics. AB - Preclinical and clinical data suggest a certain antitumor efficacy for combination of retinoids, cytokines and cytotoxic compounds. PATIENTS AND METHODS: Between November 1994 and October 1996, 38 patients with advanced squamous cell carcinoma were enrolled in a phase II study to investigate an association of low-dose all-trans-retinoic acid (tRA) (40 mg/m2/day, 84 days), interferon-alpha (IFN-alpha) (6.10(6) UI/day, 84 days s.c.) and cisplatin (40 mg/m2/day, day 1, 28 and 56, i.v.). A Pharmacokinetic evaluation was performed on 12 patients. RESULTS: The incidence of grade 3/4 hematologic toxicities was moderate (< 20%). Extra hematological toxicities were frequent but easily manageable and not life threatening. However, treatment delivery was poor since only 6 patients (16%) received full therapy. Seven objective responses were observed (21%), suggesting some degree of synergism between tRA, IFN-alpha and cisplatin. Interestingly, continuous tRA treatment in combination with IFN-alpha and cisplatin did not induce a significant decrease in plasma levels, as had been previously described. CONCLUSIONS: Regarding the short median response duration and the frequency of toxic events, this regimen should no longer be recommended in pretreated patients with advanced disease. However, the consistent response rate reported here may warrant further investigation in an early setting. PMID- 11396228 TI - Expression of endogenous lectins (galectins, receptors for ABH-epitopes) and the MIB-1 antigen in esophageal carcinomas and their syntactic structure analysis in relation to post-surgical tumor stage and lymph node involvement. AB - Squamous cell carcinomas of the esophagus, a disease with poor prognosis, are especially frequent in China and South Africa. To initiate the study of endogenous lectins in this tumor class we employed synthetic neoglycoconjugates and focused on galectins as markers. Histological sections of 43 cases of esophageal carcinomas were analyzed with labeled galectins-1 and -3 and their specific antibodies, neoglycoconjugates exposing chemically prepared histo-blood group A-, B- and H-trisaccharides and the antibody MIB-1 (Ki-67). Features of structural and numerical staining intensities determined quantitatively were correlated to clinical data sets of pTN stages, sex and age of patients. Low tumor stages (pT1/T2) were seen in 10/43 cases (23%) and 65% of the carcinomas surgically treated lacked notable lymph node involvement (pN0). The women were younger than the men (47 years versus 54 years). The proliferation activity of the tumor cells was high and amounted to 75% at average. The presence of galectin 1 and the structural entropy of distribution of staining with carrier-immobilized A-trisaccharide were associated with pN stages. These initial data indicate that distinct glycohistochemical features appear to have prognostic significance in this tumor class, adding to the emerging significance of this marker class in lung cancer. PMID- 11396230 TI - Posterior uveal malignant melanoma: temporal stability and ethnic variation in rates in Israel. AB - In a population-based study of posterior uveal malignant melanoma (755 incidence cases), the authors found a stabilized incidence in Israel from 1961-1996. Overall, Jewish immigrants to Israel had a relative rate (RR) of 2.2 [95% confidence interval [1.5-2.6] as compared to the reference population (Israeli born Jews with Israel-born parents, i.e. third generation). Whereas individuals born in Eastern Europe or the Americas experienced the highest age adjusted incidence rates per million [for example; from 1972-1996, 8.3 for those from Poland, 8.2 from Romania, 6.4 from the former Union of Soviet Socialist Republics, 7.6 from the Americas], the lowest incidence rates were observed among immigrants from Algeria-Morocco-Tunisia (rate of 2.8), Iraq (1.7), Iran (3.2). Jews born in Israel exhibited incidence patterns similar to those individuals from the place of their parent's birth; high rates were observed among individuals born of American- or European-born parents (rate of 7.2), and low rates among offspring of parents who migrated from Africa or Asia (2.6). A low incidence was found among Israeli-born Arabs (2.6 in men, and 2.0 in women). The stable differences in incidence rates, according to populations of Jews, and the persistence of these variations within the descendants of these populations suggest that underlying susceptibility states are related to an individual's origin. PMID- 11396229 TI - Lipiodol avid liver metastases from adrenocortical carcinoma; prospects for therapy? AB - A young lady with liver metastases from adrenocortical carcinoma was given a tracer dose of lipiodol into the hepatic artery and her liver metastases were shown to be extremely lipiodol avid. A therapeutic dose of lipiodol I131 was then given. There was not, however, any evidence of response to this treatment. PMID- 11396231 TI - 5-fluorouracil and leucovorin as second-line chemotherapy in carcinomas of unknown primary site. AB - Carcinomas of unknown primary (CUP) are highly malignant diseases which have poor prognosis. We report in the present study the impact on survival and toxicity of a salvage chemotherapy with 5-Fluorouracil (5-FU) and leucovorin (LV) in 25 patients with CUP who failed first-line treatment with cisplatin-based regimens. Chemotherapy was a bimonthly regimen of intravenous LV 200 mg/m2 as a 2-hour infusion followed by a 1-hour bolus 5-FU 400 mg/m2 and a 22-hour continuous infusion 5-FU 600 mg/m2 for 2 consecutive days every 2 weeks. The median number of cycles per patient was 4. Overall toxicity was mild. No objective response was observed. From the start of second-line chemotherapy, the median overall survival was 3 months. The median overall survival from diagnosis was 9 months. Second line chemotherapy with 5-FU/LV has no impact on survival in patients with CUP. PMID- 11396232 TI - Stability of aneuploid clones during oral squamous cell carcinoma metastasis. AB - BACKGROUND: The development of aneuploid clones contributes to the expression of metastatic growth properties in head and neck squamous cell carcinomas. Controversy persists as to whether aneuploid clones acquire an autonomous metastatic potential or instead activate diploid tumor cells to disseminate. PATIENTS AND METHODS: DNA flow cytometry was performed on 73 primary oral squamous cell carcinomas and their synchronous lymph node metastases as well as on 20 other patients who experienced delayed manifestation of occult lymph node involvement. RESULTS: Diploidy for both the primary and metastatic tumor was recognized in 6.5% of patients; 5.4% showed a shift from diploidy to aneuploidy while 3.2% had aneuploid tumors that associated with diploid metastases. Aneuploid clones in corresponding lesions were identified in 84.9% of cases. Despite the wide variation of individual DNA contents, 86.1% of the latter group expressed aneuploid clones with identical DNA indices in both the primary and metastatic tumors. The stability of initially established aneuploid clones was monitored even after 58 months of occult metastasis growth. CONCLUSION: Lymph node metastases preferentially originate from pre-existing aneuploid tumor clones. PMID- 11396233 TI - Clinical utility of serum HER-2/neu testing on the Bayer Immuno 1 automated system in breast cancer. AB - The clinical utility of automated serum HER-2/neu measurements in breast cancer run on the Bayer random analyzer Immuno 1 was analyzed in several steps: [a] The reference interval was determined for 242 normal healthy pre- and postmenopausal females. [b] The clinical specificity of serum HER-2/neu to separate healthy controls from 210 patients with non-malignant breast--and non-breast diseases was calculated. [c] The clinical sensitivity of cross-sectional serum HER-2/neu values for 204 patients (pts) with stage I-IV breast cancer was established. [d] Specimens from 103 stage IV breast cancer pts were tested for their parallel between serial serum HER-2/neu results and disease course. RESULTS: [a] The value of 13.03 ng/ml exceeded 95% of the results from the healthy female population. Based on the mean +2 standard deviations value of 14.7 ng/dl, the upper limit of normal was established at 15 ng/ml. [b] The specificity for benign breast diseases and other benign non-breast diseases was 98.0% and 94.6%, respectively. [c] The correlation of increased serum HER-2/neu levels and stage of breast cancer revealed the best sensitivity of 40% for stage IV disease. [4] Thirty eight (36.9%) of 103 stage IV patients had initial HER-2/neu values > 15 ng/ml, 33 of whom showed longitudinal HER-2/neu concentrations which paralleled the clinical course of the disease giving a sensitivity of 86.8%. PMID- 11396234 TI - Pancreatitis-associated protein (PAP) in patients with pancreatic cancer. AB - BACKGROUND: Pancreatitis-associated protein (PAP) is known as a marker for pancreatitis and cystic fibrosis. The aim of our study was to evaluate PAP in patients with pancreatic cancer, to assess its correlation to the extent of the disease and to compare it to CA19-9. PATIENTS AND METHODS: This prospective study comprised 75 individuals. Thirty had pancreatic cancer, 30 were healthy controls and 15 had benign lesions of the pancreas. PAP was determined by enzyme-linked immunosorbent assay. Statistical analysis was by Wilcoxon test and Spearman correlation coefficients. RESULTS: As compared to healthy individuals and using a cut-off of 18 micrograms/l corresponding to a sensitivity of 90%, the specificity of PAP for pancreatic cancer was 82.8%. PAP elevation in cancer patients could not be explained by concomitant pancreatitis (p = 0.649). PAP did not show correlation to tumour size (p = 0.14), T-stages (p = 0.706) or tumour grading (p = 0.105), but was significantly correlated to the overall extent of the disease according to the UICC stages (p = 0.002). No correlation between PAP and CA19-9 was seen. Jaundice was not found to influence PAP values (p = 0.4). CONCLUSION: Elevation of PAP in patients with pancreatic cancer is not merely explainable by concomitant pancreatitis, but seems to be due to increased PAP production by the cancer cells and is also correlated to tumour load as expressed by the UICC stages. PMID- 11396235 TI - Oral estramustine therapy in serum chromogranin A-positive stage D3 prostate cancer patients. AB - Neuroendocrine (NE) differentiation in prostate cancer (PC) has attracted much attention since it has been shown to be associated with androgen independence and bad prognoses. In this study 34 patients with hormone refractory prostate cancer and elevated Chromogranin A (CgA) serum values were treated with estramustine for 15 months. The results were evaluated by analyzing clinical data and the serially measured total PSA and Chromogranin A serotest concentration, as well as bone scan recordings. In responders to therapy a sharp decline in CgA level was recorded as well as a slight decrease in the number of metastatic lesions in the bone. However, in 6- and 12-month nonresponders, supranormal serum CgA concentrations were measured (mean 305 +/- 122 ng/ml/ and 284 +/- 101 ng/ml, respectively). Statistical significance was found between neither of these groups (p > 0.05). Mean bone lesions data were practically identical in nonresponding groups (9.6 and 9.5) but were much lower in the respective responders (4.9 and 4.1). Twelve patients were found to be nonresponders 3 months after therapy initiation (12 out of 34, 35.3%). From the remaining 22 studied subjects a positive response patients continued in 17 patients during the next 3 months or overall in a six-month period (17 out of 34, 50%, or 17 out of 22, 77.3% from 3 month responders). After 9 months of therapy, 15 responding, patients were found (15 out of 34, 44.1%, or 15 out of 17, 88.2% from 6-month responders) while, after a full year of estramustine administration, 8 patients responded to therapy (8 out of 34, 23.5%, or 8 out of 15, 53.3% from responders after 9 months of treatment). Finally, after 15 months of treatment, only 3 responding patients found (3 out of 34, 8.8% or 3 out of 8, 37.5% from responders after 12 months of treatment). Mean response periods after 12 and 15 months of estramustine treatment were 5.5 and 5.7 months, respectively. In conclusion, the reported results indicate a 12-15 month responding period for estramustine therapy in patients with a slight initial elevation in CgA serotest (up to 150 ng/ml) and less than 10 metastatic osseous lesions. Thus, serum CgA assessment has definitely found a role in monitoring PC patients with NE positive stage D3 disease. This specially holds true as the population of aged men is rapidly increasing, posing a great challenge to urologists and oncologists in treating difficult-to-manage hormone-independent prostatic carcinoma. PMID- 11396236 TI - Classification of in vivo 1H MR spectra from breast tissue using artificial neural networks. AB - BACKGROUND: The study was designed in order to investigate whether artificial neural networks could be used for analysis of in vivo magnetic resonance (MR) spectra from breast cancer patients. MATERIALS AND METHODS: In vivo 1H MR spectra with three different echo times (TE 135, 350 and 450 msec) were acquired from patients with benign and malignant breast lesions and from healthy volunteers, of whom some were breast-feeding. A spectral region (4.0-1.5 ppm) was used as input for artificial neural network analysis, for the attempted classification of the data into different groups. RESULTS: Data recorded at all three echo times were necessary to obtain the best results. Furthermore, malignant tissue was differentiated from benign tumours using this approach, whereas benign tumours were poorly separated from healthy tissue. CONCLUSION: The results presented here indicate that in vivo MR spectroscopy in conjunction with neural network analysis might be useful for the evaluation of breast lesions. PMID- 11396237 TI - Multi-center study of two dose levels of paclitaxel with carboplatin in locally advanced and metastatic non-small cell lung cancer (NSCLC). AB - BACKGROUND: The efficacy and toxicity of the combination of two cytotoxic compounds that are active as single agents in non-small cell lung cancer (NSCLC), paclitaxel (Taxol) and carboplatin (Paraplatin) was investigated in a multicenter, community-based setting. MATERIALS AND METHODS: Two consecutive cohorts of chemonaive patients with stages IIIA/B and IV NSCLC received two dose levels of paclitaxel. The first cohort received 200 mg/m2 over 3 hours (HD) and the second cohort 175 mg/m2 over 3 hours (LD) in combination with a fixed dose of carboplatin. The dose of carboplatin was calculated according to the Calvert formula with an area under the concentration versus time curve (AUC) value of 6 mg/ml/minute. The carboplatin clearance, calculated by the Chatelut formula rather than the glomerulation filtration rate (GFR) +25, was introduced into the Calvert formula. The eligibility criteria were identical for both cohorts throughout the study. Treatment was administered every three weeks. The study endpoints were response rate (RR), toxicity, time to progression (TTP) and survival (S). RESULTS: One hundred and thirty consecutive eligible patients from 36 Belgian institutions were fully evaluable for all study parameters (99 in the HD and 31 in the LD cohort). Myelosuppression was the most prominent side-effect of treatment with comparable results for both cohorts. The worst grade 3-4 leucopenia and neutropenia per patient in the HD versus LD cohort was 34.4 vs 19.3% and 59.2 vs 51.6%, respectively. 10.4% of patients in the HD cohort required hospitalisation for febrile neutropenia (6.2% with and 4.2% without documented bacterial infection), while in the LD cohort the respective figures were 13.7, 10.3 and 3.4%. The most prominent non-hematologic toxicities were alopecia and polyneuropathy, with no major difference between the HD and LD cohort (grade 2 alopecia in 78.1 vs. 83.9% and grade 3 neuropathy in 14.3 vs. 9.7%, respectively). The overall best clinical RR was 31 out of 130 (23.8%) with one complete (CR) and 30 partial responses (PR). The respective RR in the HD and LD cohort was 23.2 and 25.8%. Median TTP and S for all patients was 120 and 248 days, with no apparent difference between the HD and the LD cohort (119 and 254 versus 128 and 222, respectively). The one year survival was 34% in the HD cohort. The 95% confidence intervals for efficacy and toxicity parameters overlapped in both cohorts. CONCLUSION: In this multicenter study, the combination of paclitaxel and carboplatin produced a moderate RR of 23.8% in stages IIIA/B & IV NSCLC. The therapy was generally well tolerated at both doses of paclitaxel. Myelosuppression, neurotoxicity and alopecia were the major therapy-related side-effects. The differences between the two paclitaxel dose cohorts with respect to activity and toxicity were minimal. The use of the Chatelut formula to calculate the carboplatin clearance is feasible, but might have lead to the apparent excess in myelotoxicity in our study compared to other studies which used other methods for estimating renal function. PMID- 11396238 TI - Tumor markers in stage P1 bladder cancer. AB - BACKGROUND: The infiltration of muscularis mucosa in superficial bladder cancer has been reported to be predictive of an unfavourable course of the disease. MATERIALS AND METHODS: We studied immunohistochemically Ki-67, PCNA and p53 tumour markers in 68 P1a and Pib bladder tumours. RESULTS: A statistically significant difference (p = 0.01) was found in the distribution of grades between stages P1a and P1b, with more grade 3 and less grade 2 tumours in the latter category. Univariate analysis revealed a strong association of Ki-67 (p = 0.001) and PCNA (p = 0.032) only with stage. P53 protein expression did not have any significant association with either stage or grade. In 60 patients entered into the multivariate analysis a clearly predominant, significant effect of stage on Ki-67 (p = 0.000) was shown. CONCLUSION: Increased proliferative activity when compared to P1a is present in P1b bladder tumours, as detected by the increased expression of Ki-67 proliferating antigen. The immunohistochemical study of Ki-67 antigen may help in predicting stage P1 tumours' behaviour, even in cases where pathological distinction of P1a and P1b substages is difficult. PMID- 11396239 TI - Complete regression of primary gastric MALT-lymphoma after double eradication Helicobacter pylori therapy: role and importance of endoscopic ultrasonography. AB - The authors report on a case of complete regression of primary gastric Mucosa Associated Lymphoid Tissue MALT-lymphoma after double eradication Helicobacter pylori therapy. They analyze the diagnostic role of endoscopic ultrasonography and the therapeutic aspects, on the grounds of literature data and personal experience are analyzed. PMID- 11396240 TI - Melanoma involving the gastrointestinal tract. AB - We studied 19 patients, 12 males and 7 females of a median age of 49 years (range 26-76) with clinical manifestations of gastro-intestinal (GI) involvement from malignant melanoma. Their records were available from an electronic database of 2163 patients registered between 1977 and 1998 with the Melanoma Unit initially at Westminster and currently at Charing Cross Hospital. Time interval from initial presentation of melanoma to the development of GI involvement ranged from 6 to 183 months (median 44 months). Twelve patients had a laparotomy and in eight of these resection of metastatic sites was complete. Ten patients received systemic treatment. Median survival from GI involvement for patients who had resection of metastases was 21.5 months (range 1 to 253 months). Enterectomy, especially in the presence of solitary intestinal metastases, can palliate and may prolong survival. The role of adjuvant chemotherapy is less well defined but is worthy of consideration. PMID- 11396241 TI - Technetium-99m tetrofosmin scintigraphy for detecting malignant lymphoma. AB - The aim of this study was to assess the use of Tc-99m tetrofosmin (Tc-TF) scintigraphy for detecting malignant lymphomas. In the study, before any chemotherapy, 50 patients with malignant lymphoma underwent Tc-TF scintigraphy, which was performed 10 minutes after intravenous injection of 20 mCt Tc-TF. Tc-TF scintigraphy detected malignant lymphoma in 44 (88%) patients. However, there were no significant differences in the incidences of positive and negative Tc-TF scintigraphic results between female versus male patients, HD versus NHL patients, stage I-II versus stage III-IV patients, age > 40 years versus < or = 40 years patients and patients with B symptoms false-negative results occurred in 4 (8%) infradiaphragmatic malignant lymphoma. We conclude patients with that Tc TF scintigraphy appears suitable for detecting malignant lymphoma, especially supradiaphragmatic lesions. PMID- 11396242 TI - [Experiences with shorter surgery]. AB - Through the introduction of new computer-based forms of organisation, the hospital stay for both general and visceral surgery has been significantly reduced. A short stay ward with 15 beds and a dedicated functional unit was established in 1994 where approximately 1,000 operations take place each year with a 2.3 day average hospital stay. At this center laparoscopic cholecystectomy, laparoscopic herniorrhaphy, arthroscopy and perforating vein surgery are performed on ASA-I- and ASA-II-patients. Experience indicates that patients satisfaction has dramatically increased. This is due to a shorter waiting period, a guaranteed operation in accordance with the time scheduled, a comfortable hotel-like-atmosphere and minimal hospital stay without an increase of the perioperative risk. In this way hospitals can maximize their profits using special payment arrangements (all-inclusive prices). Even after introduction of the new DRG-systems short stay surgery will be attractive considering economic aspects. PMID- 11396243 TI - [Status of discontinuity resection in septic diverticular complications. History or a still current procedure?]. AB - Primary anastomosis is becoming increasingly favoured because of the shorter hospital stay even in emergency operations on the colon and rectum. This appears entirely justified when an objective of cost-effective medicine has been set. At our hospital between September 1985 and February 1999, 365 patients were operated on because of diverticular disease. The data from 346 of these patients were evaluated. 202 of these cases were elective; 144 were carried out on an emergency or urgent basis. In 223 cases, a primary anastomosis (Stage I and II according to Hinchey) and in 56 patients a double-barreled colostomy with a distal mucus fistula were performed. 57 patients had a too short rectum-sigma stump and were operated in two stages, according to Hartmann. Of 223 patients with a primary anastomosis, 6 (2.7%) developed an insufficiency of the anastomosis, whereby 3 (1.3%) of these patients subsequently died. Of the patients with Hartmann operation, 11 (19.3%) died due to the sequelae of peritonitis and only 2 patients died following insufficiency of the Hartmann's stump. Of the patients with a mucus fistula, 10 patients (17.9%) died as a result of complications of peritonitis. Of the 113 patients (21 died) with a discontinuity resection, 66 (71%) had their colostomy closed. There were no cases of anastomosis insufficiency and no patient died as a result of the colostomy closure. In the interest of the patients in septic diverticulitis the safest surgical procedure, the discontinuity resection, should be chosen. PMID- 11396244 TI - [Choledochus revision in the age of endoscopic papillotomy. Indications and outcome]. AB - OBJECTIVE: In the age of endoscopic papillotomy (EPT) the operative revision of the bile duct needs to be used stringently and interventional therapeutic options have to be considered primarily in the treatment of gallbladder and bile duct stone disease. PATIENTS AND METHODS: 70 patients treated by choledochus revision for cholecysto- and choledocholithiasis in our department between 1988-1999 were analyzed retrospectively for indication and results. RESULTS: ERCPs were performed in only 24 patients. In 17 patients no endoscopic papillotomy could be accomplished due to a former operation (Billroth II or Whipple-OP). 70 patients underwent a surgical bile duct exploration with insertion of a T-drainage. The postoperative surgical complication rate amounted to 36%. The most frequent complications after revision of the common bile duct were edematous pancreatitis as well as bile duct fistula. 37 of 70 patients could be reexamined 4-48 months postoperatively. A pancreatitis or stenosis of papilla Vateri were not seen in these patients. In the sonography the common bile duct was inconspicuous. CONCLUSIONS: In the therapy of bile duct stones the therapeutic splitting has become generally accepted. The patients should be submitted to a ERCP with EPT and subsequent laparoscopic cholecystectomy. Surgical revision of the common bile duct nevertheless remains an important tool in the therapy of choledocholithiasis. PMID- 11396245 TI - [Surgical therapy of cystic echinococcosis of the liver]. AB - This study analyses the results of a surgical deroofing technique that was used for the treatment of cystic echinococcosis of the liver in our institution during the last 18 years. The deroofing technique was used on 106 of 132 patients from January 1981 to December 1998. Other operative techniques (pericystectomy, hemihepatectomy) were performed on 26 patients. In 12 cases (9%) the operative procedure was completed by interventions at the extrahepatic bile ducts. 99 patients (75%) showed one or two cysts, and the remaining 33 patients (25%) had three or more cysts in the liver. 18 (14%) patients had to be reoperated on account of complications: in 5 cases a postoperative hemorrhage, in 11 cases a bile duct leakage and in 2 cases an intraabdominal abscess had to be treated operatively. The follow-up of 74 patients (81%) revealed recurrent disease in 14 (19%) cases. Surgical deroofing of cystic echinococcosis of the liver is easy to perform and in contrast to other minor surgical interventions universally applicable. The recurrence rate and the rate of postoperative complications are comparable to other minor surgical techniques. Therefore in our view this procedure represents the therapy of choice. PMID- 11396246 TI - [Diagnosis and therapy of progressive thrombophlebitis of epifascial leg veins]. AB - INTRODUCTION: The thrombophlebitis is generally regarded as a harmless disease. However, the progressive varicophlebitis represents a subgroup of thrombophlebitis in which the proximal portion of the thrombus can ascend into the deep vein system with the complication of deep vein thrombosis or pulmonary embolism. PATIENTS AND METHODS: In a period of 15 months ten patients were operated upon in whom a progressive varicophlebitis was diagnosed with color-flow duplex scanning. Eight of them were men, two were women. The average age was 56 years. RESULTS: Nine patients had an ascending thrombosis of the greater saphenous vein. One patient had the origin of the thrombus in the shorter saphenous vein. In one patient the ascending thrombosis of the greater saphenous vein was the reason for a segmental pulmonary embolism. The indication for operation was given when the proximal portion of the thrombus was within 10 cm of the confluence to the deep vein system. The operative procedure comprised the ligature of the epifascial vein. CONCLUSION: The diagnosis of thrombophlebitis should not be restricted only to clinical examination. Color-flow duplex scanning is preferred to compression venous ultrasonography. The ascending contrast venography as invasive diagnostic procedure should only be performed for the clarification of further questions. In ascending varicophlebitis ligature of the confluence from the superficial to the deep vein system is a safe procedure to avoid a progression of the disease or embolism. In case of postoperative recanalisation of the superficial varicose vein a second operation with stripping of the vein can be performed. PMID- 11396247 TI - [Arterial vascular injuries in fractures and dislocations]. AB - We analyzed reasons, numbers and results of arterial lesions accompanying fractures (n = 21) and luxations (n = 6) in a 6-year-period (1993-1998) retrospectively. Traffic accidents were in nearly 50% responsible for the injuries. 8 patients had suffered multiple injuries. In 17 patients the lower, and in 10 patients the upper extremities were affected. The vascular wall was completely disrupted or severed in 74%. In 7 cases (26%), patients had suffered blunt or indirect arterial trauma with intima- and media-lacerations due to subcapital fracture of the humerus (n = 2), fractured femoral bone (n = 1), luxation of the knee joint (n = 3) or the elbow (n = 1). The mean preoperative time period was 6 hours and 20 minutes (2 to 16 hours) in patients with complete ischaemia. Vascular reconstruction was performed by interposition of an autologous vein graft or an autologous venous bypass (n = 20), by direct reconstruction and primary suturing (n = 2), by use of a venous patch plasty (n = 2) and, in a single case, by autologous bypass procedure. In one case, a crural artery was ligated, in another case with a Mangled Extremity Severity Score (MESS) of 7 points a primary amputation of the lower leg was necessary. In 5 patients (19%) secondary amputations were performed. No patient died. The final outcome is mostly influenced by the preoperative period of ischaemia. PMID- 11396249 TI - [Singular late metastasis of renal cell carcinoma in the pancreas. An unusual pancreatic tumor]. AB - We present the rare case of a solitary pancreatic metastasis of renal cell cancer being the only manifestation of tumordissemination. In 1993 the patient underwent a transperitoneal nephrectomy for removal of a clear cell renal cancer. In 1998 during follow up we detected a pancreatic tumor which was treated by pancreas resection, splenectomy and resection of the colon transversum. The histology revealed a metastasis of the previously resected renal cancer. The postoperative recovery was impaired by the development of an abcess in the former pancreatic region. During the first year of follow up we did not find any signs for local or distant tumor recurrence. Pancreas resection adapted to the location of the tumor is the treatment of choice for isolated solitary late metastases of renal cell cancer. PMID- 11396248 TI - [Prevention of thromboembolism in spinal fractures with spinal cord injuries. Standard heparin versus low-molecular-weight heparin in acute paraplegia]. AB - In a prospective, randomized, open study, the therapeutic efficacy of a long-term prophylaxis with standard heparin (SH) was compared with that of low-molecular weight heparin (LMWH) Dalteparin in 166 patients who had spinal fractures with spinal cord injury. 86 patients were treated with SH 2 x 7500 U s.c. and 80 patients were treated with LMWH 1 x 5000 anti-Xa U s.c. once daily. The screening was implemented by daily bedside-examination. In case of clinical thromboembolism symptoms patients had confirmatory venography or lung scans. In the SH-group 12 (14.0%) patients had deep vein thrombosis and in the LMWH-group 6 (7.5%). Pulmonary embolism was detected two times in the SH-group (2.33%) and only one time in the LMWH-group (1.25%). A significant difference could not be shown, but is descriptive evident. PMID- 11396250 TI - [Aneurysm of the femoral vein as differential diagnosis of femoral hernia]. AB - Venous aneurysms are rare, especially those of the femoral vein. Groin hernias and femoral hernias are in contrast to this more frequent enteties. The literature includes only six cases of aneurysms of the femoral vein. We report on a 61-year-old patient who underwent an operation for femoral hernia. An aneurysm of the femoral vein was found intraoperatively and directly resected. The differential diagnosis of femoral hernias and the following diagnostic and therapeutic strategies are described. PMID- 11396251 TI - [Early abdominal manifestation of neurofibromatosis type ! in a 12-year-old boy]]. AB - Neurofibromatosis type 1 (NF 1) is a neuroectodermal tumor in which abdominal manifestations occur in about 25% of cases in addition to the typical cutaneous lesions. A child with a severe form of an abdominal manifestation of NF 1 was treated in the Hanover Medical School by tumorresection. Pre-operative diagnosis using CT, angiography and ultrasound showed an enormous abdominal tumor constricting the mesenteric vessels. The tumor and the affected portion of the small intestine (150 cms) were removed surgically. No tumor recurrence was observed nine months after operation. Surgical removal is the only currently available therapy for treating tumors of this size. PMID- 11396252 TI - [Experimental validation of DXA and MRI-based bone density measurement by ash method]. AB - Different methods have been established for bone density measurements such as dual energy X-ray absorptiometry (DXA), quantitative computertomography (QCT), and scintigraphy (VQ-Scan). There are hints that magnetic resonance imaging (MRI) might become a new option for the evaluation of bone density. The aim of this study was to investigate correlations between MRI vs. DXA and MRI vs. mineral content of lumbar vertebrae. Data were obtained from ten lumbar vertebral bodies of cattles. The T-1 MRI-sequences SE, PS, and the T-2 Sequence STIR were used for analysis. Total pixel numbers and a pixel per area ratio were determined. Values were compared to DXA-measurements, to the wet weight, and to separated measurements of the spongious, trabecular, and total mineral content of the vertebral body after ashing. We found correlations between DXA (g/vertebral body) vs. mineral content by ash-method (0.918; p < 0.01), DXA vs. MRI (SE-sequence) ( 0.872; p < 0.01), and MRI (SE-sequence) vs. mineral content (0.775; p < 0.01). No correlations were found between PS- or STIR-sequences and the ash-method. This study shows that the determination of the bone mineral content of vertebrae is possible applying MRI in the T1-weighted SE-sequence. Without radiation, the MRI provides additionally early detection of trabecular lesions, fractures, and deformities at the spine, without other diagnostic procedures becoming necessary. PMID- 11396253 TI - [Experimental study of antiseptic (tauroline, taurolidine) and antibiotic (sulmycin implant) drugs in vascular prosthesis infections in a standardized infection model]. AB - It was the aim to examine whether local application of antiseptic and antibiotic substances is an effective treatment of vascular graft infection. MATERIAL AND METHODS: 19 pigs with a bodyweight between 20 and 30 kg were assigned to three different groups. Group I: control (6), group II: local treatment with Sulmycin implant, group (6) III: local treatment with Taurolin (Taurolidine) (7). An unprotected vascular graft was inserted in the right femoral artery of all pigs. After finishing the proximal and distal anastomosis and prior to closure of the incision, the vascular grafts were contaminated locally with 2 x 10(7) CFU/ml Staphylococcus aureus ATCC 29213. Seven days later all animals received another unprotected vascular prosthesis with or without additional treatment according to groups I, II, III. 28 days after primary operation the animals were euthanized and the grafts harvested. The specimens were examined for signs of infection by histology and microbiology. RESULTS: After the primary operation all animals presented with infected vascular prosthesis. At termination of the trial on day 28 all grafts of group I were contaminated, 5 out of 6 grafts in group II, and 5 out of 7 in group III presented with infected grafts. There was no significant statistical difference between the groups. Infection could not be prevented by the antimicrobial agents used. The primary infecting organism Staphylococcus aureus, however, was eliminated in all cases. CONCLUSIONS: Both antimicrobial substances examined were not effective in the treatment of vascular graft infection, but might be used as adjuvant therapy of vascular graft infection, whereby Sulmycin implant seems to be more effective regarding the incorporation of the prosthesis. PMID- 11396254 TI - [Our surgical heritage. The surgical schools in Magdeburg]. AB - During the previous two centuries several leading personalities have worked in the two major surgical hospitals of the city of Magdeburg. This retrospective view remembers a few of these surgeons and their merits for patients, education and scientific life. In the hospital "Krankenhaus Altstadt Magdeburg" the work of surgeons dates back to 1827 with the foundation of a surgical hospital including a teaching facility for surgeons as "Wundarzte II. Klasse". Geheimrat Dr. W. Hagedorn has been the outstanding personality of the 19th century whereas Prof. Dr. Max Biebl is regarded the same for the 20th century. The hospital "Sudenburger Krankenhaus" was founded in 1891. Since 1954 it developed by Prof. Dr. W. Lembcke into an academic teaching hospital. As "Center of Surgery" it is today part of the medical faculty of the university of Magdeburg. PMID- 11396255 TI - [Therapy of inguinal hernia (1)]. PMID- 11396256 TI - [Fecal incontinence: various causes and treatments]. AB - Three patients, two women aged 73 and 54 years and one man aged 46 years, had faecal incontinence for several years. All three patients had sphincter defects; two were managed conservatively and one was treated surgically with acceptable results. Of the many possible causes of faecal incontinence, a disturbed defecation pattern and obstetric trauma are most frequently encountered. After medical history and physical examination, anal endosonography is important in demonstrating or excluding an anal sphincter defect. Conservative treatment by means of a fibre-enriched diet, laxatives, pelvic floor exercises or daily enemas may provide a socially acceptable balance for the patient. When conservative management fails and a sphincter defect is present, sphincteroplasty is indicated. PMID- 11396258 TI - ['Inflammometry' with nitric oxide in exhaled air: a new test for lung diseases]. AB - The gas nitric oxide (NO) is produced in increased amounts in certain types of inflammatory responses and its presence in exhaled air can be demonstrated. The nitric oxide fraction in exhaled air (FeNO) is elevated in patients with asthma and lowered in the case of several other lung diseases such as cystic fibrosis and primary ciliary dyskinesia. The FeNO can be quickly measured in a non invasive and reproducible manner: on-line if the patient (adult or child), having taken a deep breath in, breathes out with a low flow rate into the NO measuring device or off-line if the expired air is collected in an NO inert reservoir. Confounding factors are contamination of inhaled air with ambient NO and contamination of exhaled air with NO that has been produced in the paranasal sinuses and the nose. The possible applications of FeNO measurement as a new lung function test include diagnostic tests for chronic respiratory symptoms and the possible guidance of anti-inflammatory therapy for asthma and, perhaps, other respiratory disorders. PMID- 11396259 TI - [Treatment of pain in cancer with systemically administered opioids]. AB - The World Health Organization guidelines for cancer pain relief have been proven efficacious in 90% of the patients with cancer pain. The patient's self-report of pain is the focus of treatment. When initiating treatment, controlled-release preparations of opioids are generally favoured, and are combined with immediate release morphine to prevent or treat 'breakthrough' pain and to enable the optimum opioid dosage to be calculated. (Breakthrough pain is a transient increase in pain in a patient who has stable, persistent pain treated with opioids.) In patients with an unfavourable balance between analgesia and side effects, the following strategies may be useful, together with appropriate treatment of the side effects: Sequential opioid trials (so-called opioid rotation) is an approach which is effective in 50-70% of the patients. Changing the route of opioid administration is successful in 70-95% of the patients. When selecting an invasive technique, continuous subcutaneous infusion is medically preferred. Spinal analgesia is an alternative. Knowledge of the relative potency of opioid drugs and of their biologic availability is needed to guide changes in drugs or routes of administration. PMID- 11396260 TI - [Diagnostic image (38). Allergic contact dermatitis]. AB - A 21-year-old woman who presented with acute eczema was diagnosed with allergic contact dermatitis from colofonium in tape that she used after a sprain of her left ankle. PMID- 11396261 TI - [From gene to disease; from homocysteine to hyperhomocysteinemia]. AB - Homocysteine is a sulfhydryl containing amino acid which is produced as an intermediate product in the metabolism of the essential amino acid methionine. Apart from environmental factors such as the intake of folate and other B vitamins, the level of homocysteine in the blood is determined by the genetically based activities of several enzymes involved in the methionine or folate cycle. The well-known congenital defect homocystinuria is due to homozygosity for mutated cystathionine beta synthase. It is characterized by severe hyperhomocysteinemia, which leads to arterial and venous disease at a very young age. Mild to moderate hyperhomocysteinemia, due to either heterozygosity for severe mutations in the genes of enzymes involved or based upon homozygosity for more mild mutations, has also been recognized as a risk factor for vascular disease in the last decade. However, ongoing clinical intervention studies still need to demonstrate a casual role of mildly increased homocysteine levels in vascular disease. PMID- 11396262 TI - [Drug utilization by the elderly with depressive symptoms]. AB - OBJECTIVE: Obtain a picture of non-psychiatric medication use and depressogenic medication use by elderly people suffering from depression. DESIGN: Cross sectional, self-reported by respondents. METHOD: Information about medication usage and the presence of chronic diseases was obtained from 183 depressive respondents and 96 control respondents. The respondents were aged 55 years or over. They were recruited between April 1996 and May 1998 in the provinces Groningen, Drenthe and Friesland, the Netherlands. Recruitment was via Regional Institutes for Outpatient Mental Health Care (RIAGGs), a psychiatric outpatient clinic, general practitioners participating in the 'Groningen longitudinal ageing study' (GLAS), a survey under the elderly within the general population, and screening with the 'Geriatric depression scale' (score > or = 6) in the GLAS main file. RESULTS: Respondents with subsyndromal depression recruited via screening were prescribed more depressogenic medication (mean: 3.26 drugs per patient, of which 45.6% depressogenic) than the control group (respective figures: 1.78; 26.1%) and the depressive groups recruited via general practitioners (2.77; 31.8%) or those recruited via RIAGGs/psychiatric outpatient clinic (2.60; 38.3%). PMID- 11396263 TI - [Autosomal dominant cerebellar ataxias in the Netherlands: a national inventory]. AB - OBJECTIVE: To provide a comprehensive estimate of the number of Dutch autosomal dominant cerebellar ataxias (ADCA) families and patients and thus estimate the minimal prevalence of ADCA in the Netherlands. Furthermore, to observe the relative frequency of SCA mutations and to study genotype-phenotype correlations. DESIGN: Descriptive study and prevalence computation. METHODS: Genotyping was based on cytosine-adenine-guanine (CAG)-repeat expansion detection in the SCA1, SCA2, SCA3, SCA6 and SCA7 genes. We analysed the results of SCA mutation analysis with respect to the number of genotyped families, gene carriers, and clinically affected individuals per SCA locus, as well as individual repeat length and age of onset. Parent-offspring couples were studied for anticipation. The minimal prevalence was extrapolated, based on the observation that at least 36% of ADCA families cannot be genotyped. RESULTS: Per May 1st 2000, 137 Dutch ADCA families were genotyped (SCA1: 15 families; SCA2: 14; SCA3: 64; SCA6: 28; SCA7: 16) and 382 affected individuals had been identified within these families. The extrapolated minimal prevalence was 2.8 per 100,000. All lengths of expanded trinucleotide repeats identified confirmed earlier results regarding pathogenic sizes. Anticipation with an associated increase of repeat length was observed in SCA2 and SCA3 families. The length of the trinucleotide repeat accounted for 65% of the variance in age of onset in SCA3. PMID- 11396264 TI - [Macroprolactinemia, a benign form of hyperprolactinemia]. AB - In a 34-year-old woman with primary subfertility, a strongly increased serum concentration of prolactin was found in combination with normal levels of oestradiol, which is an indication for the presence of prolactin forms without clinical effect. She appeared to have macroprolactinaemia, i.e. the presence of circulating large forms of prolactin (up to > 100 kDa), which may be detected by the immunoassays currently used. They give no clinical signs or symptoms. Diagnosing macroprolactinaemia means that further diagnostic tests for hypopituitarism abnormalities using MRI need not be carried out and unjustified treatment of otherwise healthy persons may be prevented. PMID- 11396265 TI - [Psychiatric medical history of Vincent van Gogh]. PMID- 11396266 TI - Contrast-enhanced transcranial Doppler sonography in patients with acute cerebrovascular diseases. AB - The aim of the study was to evaluate the diagnostic potential of galactose-based microbubble suspension (Levovist) in patients with acute cerebrovascular disease and inadequate transtemporal acoustic window, when examined by transcranial Doppler (TCD). We studied 10 patients with either transient ischemic attack (no. = 3) or stroke (no. = 7). Inadequate transtemporal acoustic window was unilateral in 3 patients and bilateral in the remaining 7 patients. Signals from middle, anterior, and posterior cerebral arteries (MCA, ACA, PCA) were recorded after injecting Levovist 300 mg/ml. Six patients needed 3 injections of Levovist, 1 patient two, and 3 patients one. Mean +/- SD duration of optimal signal enhancement was 175.2 +/- 53.2 s, range 70-290 s. Doppler waveform analysis was possible in 14 (82.3%) MCA, 11 (65%) ACA, and 9 (53%) PCA. Levovist improved the reliability of TCD in patients with acute cerebrovascular disease and insufficient transtemporal insonation. PMID- 11396267 TI - Treatment of Parkinson's disease by cell transplantation. PMID- 11396268 TI - Multipotent neuroepithelial stem cell differentiation: some caveats for future clinical applications. PMID- 11396269 TI - Mismatch negativity and personality traits in chronic primary insomniacs. AB - The thalamo-(fronto)cortical circuit is involved in sleep regulation, and its dysfunction might contribute to the pathophysiology of chronic primary insomnia. To obtain more evidence of the involvement of the circuit, we studied 23 patients with chronic primary insomnia and 28 healthy volunteers via the assessment of mismatch negativity (MMN) elicited by tone intensity deviance, and of personality traits measured by Zuckerman's Sensation Seeking Scales, and Zuckerman-Kuhlman's Personality Questionnaire. In insomniacs, MMN amplitude at Fz was significantly larger; Depression, which was measured by Plutchik-van Praag's Depression Inventory, and Neuroticism-anxiety and Impulsivity scores were higher, while the Thrill and adventure seeking score was lower; MMN amplitude was positively correlated with Depression and with Impulsivity. In healthy subjects, MMN amplitude at Fz was positively correlated with Neuroticism-anxiety, but negatively with Experience seeking. The larger MMN and distinct personality traits suggest a hyperactivity in the thalamo-(fronto)cortical neuronal circuit in insomniacs, which is probably the result of weak thalamic gating mechanisms, or an imbalance of several neurotransmitter systems. PMID- 11396270 TI - Monogenic Parkinsonisms and the genetics of Parkinson's disease. PMID- 11396271 TI - Neurotrophic factors in the therapy of Parkinson's disease. PMID- 11396272 TI - Clinical and electrophysiological effects of apomorphine in Parkinson's disease patients are not paralleled by amino acid release changes: a microdialysis study. AB - We performed a microdialysis investigation of extracellular amino acid (glutamate and GABA) concentrations during sterotaxic neurosurgery (the implantation of permanent electrodes in the internal globus pallidus (GPi) or subthalamic nucleus (STN) for deep brain stimulation in advanced Parkinson's disease (PD) patients, after prolonged therapy wash-out). Electrophysiological single unit recordings and perioperative clinical status assessments were also performed. Amino acid levels were measured in the GPi and GPe (external globus pallidus) of three PD patients and in the STN of another three PD patients. Stable basal release values of the examined amino acids were obtained within one hour. In clinical "off" state, the basal levels of GABA in the GPi were double those in the GPe in all the three patients. This finding could represent a biochemical marker for GPi target identification in PD surgery. Acute subcutaneous apomorphine administration induced electrophysiological changes and clinical amelioration but did not change amino acid concentrations. This result could be due to methodological limitations of the microdialysis technique. Alternatively, it could suggest that the clinical effects of acute apomorphine might also be mediated by direct activation of dopaminergic receptors located in the output nuclei. PMID- 11396273 TI - Deep brain stimulation in Parkinson's disease. PMID- 11396274 TI - Neurochemistry of SOD1 and familial amyotrophic lateral sclerosis. PMID- 11396275 TI - Optimal use of antibiotics for intubation-associated pneumonia. PMID- 11396276 TI - Pharmacodynamics, a tool for a better use of antibiotics? PMID- 11396277 TI - Filling the intensive care memory gap? PMID- 11396278 TI - On the need for observational studies to design and interpret randomized trials in ICU patients: a case study in stress ulcer prophylaxis. PMID- 11396279 TI - Impact of appropriateness of initial antibiotic therapy on the outcome of ventilator-associated pneumonia. AB - OBJECTIVES: To evaluate the impact of appropriate initial antibiotic therapy (AB) on the outcome of ventilator-associated pneumonia (VAP). DESIGN: Retrospective study (1992-97). PATIENTS AND METHODS: Episodes of VAP diagnosed on both clinical and microbiological criteria after > or = 48 h of mechanical ventilation (MV). Initial AB was considered appropriate when all significant organisms were susceptible to at least one of the antibiotics started after distal bronchial sampling. Antibiotic treatment was modified within 48 h when susceptibility testing was available. Outcome was recorded at the ICU and hospital discharge. RESULTS: One hundred and eleven patients were included (SAPS II = 48 +/- 18, age = 62 +/- 14 years, mean duration of MV before VAP = 12 +/- 9 days). Initial AB was appropriate in 55 patients (49.5%). No difference between appropriate initial AB and inappropriate initial AB was found concerning severity indices at the time of VAP diagnosis. ICU length of stay was shorter with appropriate initial AB than with inappropriate initial AB for survivors (12 +/- 11 days vs 20 +/- 24 days, P = 0.01). Crude hospital mortality tended to be lower with appropriate initial AB than with inappropriate initial AB (47.3% vs 60.7%, odds ratio = 1.72, 95% CI = 0.81-3.7). Relative crude mortality reduction with appropriate initial AB was 22%, 95% CI = -10% to 45%. CONCLUSION: Inappropriate initial AB of VAP during the first 48 h increased ICU length of stay after VAP diagnosis and tended to increase crude hospital mortality despite equal severity of illness at the time of VAP diagnosis, when compared to appropriate initial AB in a population of 111 ICU patients. PMID- 11396280 TI - Low cefpirome levels during twice daily dosing in critically ill septic patients: pharmacokinetic modelling calls for more frequent dosing. AB - OBJECTIVES: To measure plasma levels and pharmacokinetics of cefpirome in critically ill septic patients with normal renal function. To use the pharmacokinetic model to simulate alternate dosing regimens and identify those that predict sustained levels. DESIGN AND SETTING: A prospective, open-label, descriptive study in a 22-bed, multidisciplinary, adult ICU in a university affiliated, tertiary referral hospital. PATIENTS: Twelve adults with normal renal function on enrollment and with suspected or documented sepsis in whom cefpirome was judged to be the appropriate therapy by the managing clinician. INTERVENTIONS: Cefpirome 2 g was infused intravenously over 3 min every 12 h. Timed blood samples were taken prior to and during two dosing intervals. Urine was collected for creatinine clearance determination. MEASUREMENTS AND RESULTS: Two patients were non-evaluable due to renal dysfunction post-enrollment. The median cefpirome trough level was 1.1 mg/l (range 0.5-8.1 mg/l) after the initial dose and 1.4 mg/l (range < 0.5-15.9 mg/l) at 'steady state'. The volumes of distribution and elimination half-lives were greater and more variable than in healthy volunteers. Pharmacokinetic simulation predicted that more frequent bolus dosing, increased doses and continuous infusions would result in concentrations greater than 4 mg/l for over 60% of the dosing interval for all patients. CONCLUSIONS: Cefpirome 2 g twice daily produced low plasma troughs in a number of patients. Our data suggest that this drug regimen may be inadequate for successful treatment of difficult-to-treat infections in critically ill patients with normal renal function. PMID- 11396281 TI - Pharmacokinetics of levofloxacin during continuous veno-venous hemofiltration. AB - OBJECTIVE: To study the effect of continuous veno-venous hemofiltration (CVVHF) on the pharmacokinetics of levofloxacin in critically ill patients with acute renal failure. DESIGN: Open-label study. SETTING: Anesthesiology ICU, University Hospital of Regensburg. PATIENTS: Six critically ill patients treated with CVVHF because of acute renal failure needing antimicrobial therapy. INTERVENTIONS: Levofloxacin i.v. 250 mg qd with a starting dose of 500 mg. CVVHF with the following characteristics: hemofilter AN69 hollow fibers of 0.90 m2 area, blood flow 150 ml/min, ultrafiltrate flow 1.3 l/h, filtrate substitution in post dilution mode. MEASUREMENTS AND RESULTS: The plasma pharmacokinetics and clearance of levofloxacin by hemofiltration were established on day 1 and day 4-6 of treatment. Levofloxacin was determined by high-performance liquid chromatography (HPLC). Mean (range) peak plasma concentrations after levofloxacin 500 mg single dose (s.d.) and 250 mg multiple dose (m.d.) were 6.4 (2.7-9.4) and 8.2 (4.7-10.3) mg/l, trough levels 2.7 (1.4-5.0) and 2.9 (1.7-3.9) mg/l, half life 28 (19-38) and 22 (17-31) h, volume of distribution 1.2 (0.72-1.6) l/kg and 0.91 (0.52-2.0) l/kg, respectively. The mean sieving coefficient was 0.96 (0.79 1.09), mean total clearance 47 (20-89) ml/min, and mean clearance by hemofiltration 21 (13-27) ml/min, respectively. CONCLUSIONS: A dosage schedule of levofloxacin 250 mg qd with a 500 mg loading dose seems appropriate for anuric patients during CVVHF. Sufficiently high steady-state concentrations of levofloxacin were achieved after the first dose. Undesired accumulation of levofloxacin was not observed. PMID- 11396282 TI - Prophylactic hemofiltration in severely traumatized patients: effects on post traumatic organ dysfunction syndrome. AB - OBJECTIVE: To evaluate the effects of prophylactic veno-venous hemofiltration (CVVH) in the absence of renal failure on multiple organ dysfunction syndrome after severe multiple trauma. DESIGN: Prospective, randomized study. SETTING: Intensive care unit (ICU) in a university hospital. PATIENTS: Twenty-four patients with severe multiple trauma (injury severity score > or = 27), no renal failure on admission and no contraindication for moderate heparinization. INTERVENTIONS: Twelve patients received conventional treatment while 12 patients were treated additionally with isovolemic CVVH for 5 days starting within 24 h following trauma. Signs of organ dysfunction were assessed daily including monitoring of systemic hemodynamic by means of pulmonary artery catheterization during the first 5 days after trauma. MEASUREMENTS AND MAIN RESULTS: Prophylactic CVVH did not affect the overall severity of organ dysfunction as assessed by MOF or APACHE II scores. However, the pattern of impaired organ systems was influenced by CVVH: while the post-traumatic decrease in platelet count in patients subjected to CVVH was more pronounced than in controls (e.g. day 4: control: 115,080 +/- 15,087, CVVH: 57,383 +/- 4,201 microliters-1; p < 0.05) the development of hyperdynamic circulatory failure was simultaneously attenuated, as reflected by a limited increased in cardiac output and an attenuated decrease in systemic vascular resistance and oxygen extraction ratio (e.g. systemic vascular resistance on day 4: control: 624.3 +/- 46.17, CVVH: 842.7 +/- 79.24 dyn.s.cm-5; p < 0.005). CONCLUSION: CVVH blunts the cardiovascular response to multiple trauma and increases tissue oxygen extraction. However, the concomitant decrease in platelet counts represents a limitation for the use of prophylactic CVVH in surgical patients. PMID- 11396283 TI - Effects of bronchoalveolar lavage volume on arterial oxygenation in mechanically ventilated patients with pneumonia. AB - OBJECTIVE: To assess the effect of bronchoalveolar lavage (BAL) volume on arterial oxygenation in critically ill patients with pneumonia. DESIGN: Randomized clinical comparison. SETTING: Six-bed respiratory intensive care unit of a 850-bed tertiary care university hospital. PATIENTS: Thirty-seven intubated and mechanically ventilated patients with clinical suspicion of pneumonia. INTERVENTIONS: Bronchoscopically guided protected specimen brush (PSB) followed by either a "high volume" BAL (n = 16, protected catheter, mean volume: 131 +/- 14 ml) or a "low volume" BAL (n = 21, protected double-plugged catheter, 40 ml volume for all patients). MEASUREMENTS: Arterial oxygen tension/fractional inspired oxygen (PaO2/FIO2) and mean arterial pressure (MAP) before and up to 24 h after the intervention. Bacterial growth in quantitative cultures. Analysis of variance for repeated measurements with inter-subject factors. RESULTS: All patients showed a lower PaO2/FIO2 ratio and higher MAP after the diagnostic procedure, without differences between the study arms (p = 0.608 and p = 0.967, respectively). Patients with significant bacterial growth (p = 0.014) and patients without preemptive antibiotic (p = 0.042) therapy showed a more profound and longer decrease in arterial oxygenation after the diagnostic procedure. CONCLUSIONS: A decrease in the PaO2/FIO2 ratio was observed in all patients after a combined diagnostic procedure, independent of the BAL volume used. A significant bacterial burden recovered from the alveoli and no preemptive antibiotic therapy were associated with a larger and longer-lasting decrease in arterial oxygenation. PMID- 11396284 TI - Trauma stimulates the synthesis of Gc-globulin. AB - OBJECTIVE: Actin is the dominating intracellular protein and is released to the circulation after tissue injury. Gc-globulin is one of the plasma proteins responsible for removal of actin from the circulation. Recent studies have shown that the level of Gc-globulin is reduced shortly after trauma. Serial changes in Gc-globulin after severe injury have not been studied so far and could provide additional information about the role of Gc-globulin in the pathophysiological response to trauma. DESIGN: Prospective, observational. SETTING: Surgical intensive care unit in a university hospital. PATIENTS: Thirty-eight patients were included in the study: 12 women and 26 men with a median age of 38 years (range 19-86) and a median Injury Severity Score (ISS) of 18 (range 6-45). Seven patients died, on day 5, 8, 8, 10, 10, 13 and 21, respectively. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: The serum concentration of Gc-globulin (Gctotal) and the percentage of Gc-globulin bound to actin (Gc%complexed) were measured daily for 1 week using rocket immunoelectrophoresis. Concentrations of free Gc-globulin (Gcfree) and Gc-globulin bound to actin (Gcbound) were calculated from these analytical results. The concentration of Gctotal and Gccomplexed correlated significantly (r = -0.99, p < 0.001) throughout the time period. After day 3 levels of Gc%complexed normalised, whereas levels of Gctotal continued to increase above control values. The concentrations of Gctotal and Gcfree were significantly lower in non-survivors compared to survivors; p = 0.005 and p = 0.03, respectively. This was combined with an inverse correlation of Gcbound between these two groups (r = -0.73; p = 0.04). CONCLUSIONS: Severe injury results in a prolonged load on the extracellular actin scavenger system; more pronounced in patients who do not survive. Gc-globulin displays characteristics of an acute phase reactant, with supra-normal serum levels 1 week after severe injury. Serial measurements of Gc-globulin after trauma could prove to be a method of early identification of patients with increased risk of mortality. PMID- 11396285 TI - Intensive care management of head-injured patients in Europe: a survey from the European brain injury consortium. AB - OBJECTIVES: (a) to describe current practice in the monitoring and treatment of moderate and severe head injuries in Europe; (b) to report on intracranial pressure and cerebral perfusion pressure monitoring, occurrence of measured and reported intracranial hypertension, and complications related to this monitoring; (c) to investigate the relationship between the severity of injury, the frequency of monitoring and management, and outcome. METHODS: A three-page questionnaire comprising 60 items of information has been compiled by 67 centres in 12 European countries. Information was collected prospectively regarding all severe and moderate head injuries in adults (> 16 years) admitted to neurosurgery within 24 h of injury. A total of 1005 adult head injury cases were enrolled in the study from 1 February 1995 to 30 April 1995. The Glasgow Outcome Scale was administered at 6 months. RESULTS: Early surgery was performed in 346 cases (35%); arterial pressure was monitored invasively in 631 (68%), ICP in 346 (37%), and jugular bulb saturation in 173 (18%). Artificial ventilation was provided to 736 patients (78%). Intracranial hypertension was noted in 55% of patients in whom ICP was recorded, while it was suspected in only 12% of cases without ICP measurement. There were great differences in the use of ventilation and CPP monitoring among the centres. Mortality at 6 months was 31%. There was an association between an increased frequency of monitoring and intervention and an increased severity of injury; correspondingly, patients who more frequently underwent monitoring and ventilation had a less favourable outcome. CONCLUSIONS: In Europe there are great differences between centres in the frequency of CPP monitoring and ventilatory support applied to head-injured patients. ICP measurement disclosed a high rate of intracranial hypertension, which was not suspected in patients evaluated on a clinical basis alone. ICP monitoring was associated with a low rate of complications. Cases with severe neurological impairment, and with the worse outcome, were treated and monitored more intensively. PMID- 11396286 TI - Risk factors for prolonged ventilation after cardiac surgery using APACHE II, SAPS II, and TISS: comparison of three different models. AB - OBJECTIVE: To identify the risk for prolonged mechanical ventilation in cardiac surgical patients. DESIGN: Prospective study with retrospective combination of a second database. PATIENTS: Six hundred and eighty-seven patients after cardiac surgery over a period of 12 months. MEASUREMENTS: Demographic data were recorded preoperatively, and surgical procedures intraoperatively using a surgical database designed for quality control. Length of ICU and hospital stay, and hospital outcome were recorded. Severity of illness was assessed daily using APACHE II, SAPS II, and Organ Failure Score. Intensity of treatment and nursing care was monitored by means of the Therapeutic Intervention Scoring System (TISS). Univariate and multivariate analyses were performed using logistic regression. The predictive value of the identified variables was tested by the Wilcoxon test using the receiver operating characteristic curve. MAIN RESULTS: Sixty-two patients (9.0%) were ventilated for > 48 h and accounted for 42.8% of the total costs in the ICU. The pre- and intraoperatively collected data produced a model with weak predictive capacity for prolonged ventilation [area under curve (AUC) 73.22 and 71.08, respectively]. The use of TISS and SAPS postoperatively resulted in an effective model of prediction (AUC 93.76). Adding the occurrence of reoperation, reintubation, emergency transfusion, intraaortic balloon pumping, and need for total parenteral nutrition to the model further improved its predictive capacity (AUC 94.74). CONCLUSIONS: The present results strongly suggest that data collected postoperatively using established scoring systems as well as documented events of high clinical impact for risk assessment and quality control are reliable predictors of prolonged ventilation. PMID- 11396287 TI - Hepatic oxygen exchange and energy metabolism in hyperdynamic porcine endotoxemia: effects of the combined thromboxane receptor antagonist and synthase inhibitor DTTX30. AB - OBJECTIVE: We compared the effects of thromboxane receptor antagonist and synthase inhibitor DTTX30 on systemic and liver blood flow, oxygen (O2) exchange and energy metabolism during 24 h of hyperdynamic endotoxemia with untreated endotoxemia. DESIGN: Prospective, randomized, experimental study with repeated measures. SETTING: Investigational animal laboratory. SUBJECTS: Twenty-seven domestic pigs: 16 during endotoxemia with volume resuscitation alone; 11 with endotoxemia, volume resuscitation and treatment with DTTX30. INTERVENTIONS: Continuous infusion of Escherichia coli lipopolysaccharide (LPS) for 24 h together with volume resuscitation. After 12 h of endotoxemia, DTTX30 was administered as a bolus of 0.12 mg kg-1 followed by 12 h continuous infusion of 0.29 mg kg-1 per h. MEASUREMENTS AND RESULTS: DTTX30 effectively counteracted the endotoxin-associated increase in TXB2 levels and increased 6-keto-PGF1 alpha with a significant shift of the thromboxane/prostacyclin ratio towards predominance of prostacyclin. DTTX30 prevented the significant progressive endotoxin-induced decrease of mean arterial pressure (MAP) below baseline while maintaining cardiac output (CO), and increased the fractional contribution of liver blood flow to CO without an effect on either hepatic O2 delivery or O2 uptake. The mean capillary hemoglobin O2 saturation (HbO2) on the liver surface and HbO2 frequency distributions remained unchanged as well. CONCLUSIONS: DTTX30 significantly attenuated the endotoxin-induced derangements of cellular energy metabolism as reflected by the diminished progressive decrease in hepatic lactate uptake rate and a blunted increase in hepatic venous lactate/pyruvate ratios. While endotoxin significantly increased the endogenous glucose production (EGP) rate, EGP returned towards baseline levels in the DTTX30-treated group. Thus, in our model DTTX30 resulted in hemodynamic stabilization concomitant with improved hepatic metabolic performance. PMID- 11396288 TI - Use of a personal diary written on the ICU during critical illness. AB - OBJECTIVE: To explore the use of a diary as an aid in debriefing patients and relatives following critical illness. DESIGN: Observation study. SETTING: Intensive care unit of a 500-bed hospital. PATIENTS AND PARTICIPANTS: Fifty-one critically ill patients and their relatives. METHOD: A daily account of the patient's progress was written in everyday language by nursing staff, photographs were added as necessary. The booklet was given to the patient or a relative at a follow-up appointment 2 weeks after discharge from the unit. A standard questionnaire was mailed 6 months later, responses were analyzed by an independent observer. MEASUREMENTS AND RESULTS: All diaries had been read by survivors (n = 41) or relatives (n = 10), 51% of the diaries had been read more than 10 times. Comments in the questionnaires were graded as very positive (39%), positive (28%) and neutral (33%). CONCLUSIONS: A detailed narrative of the patient's stay is a useful tool in the debriefing process following intensive care. PMID- 11396289 TI - Procalcitonin does not influence the surface expression of inflammatory receptors on whole blood leukocytes. AB - The plasma levels of procalcitonin (PCT) are increased in patients with severe bacterial infections. Its cellular origin and potential pathophysiological function in sepsis is, however, unclear. White blood cells have recently been described to express both PCT mRNA and protein. The aim of this study was to determine whether PCT has any influence on the surface expression of receptors, relevant in inflammation, on human whole blood leukocytes under normal and septic conditions. Venous blood from healthy donors was incubated with PCT (40 ng/ml or 1200 ng/ml) alone or in combination with lipopolysaccharide (LPS, 10 ng/ml) or peptidoglycan (PepG, 10 micrograms/ml) for 6 h. The surface expression of CD14, CD54, CD64, CD80, CD86 and HLA-DR was determined by flow cytometry. We could not detect any influence of PCT on the expression of these receptors. Further studies on potential effects on other cell types during infection seem warranted. PMID- 11396290 TI - Frequency of transient streptococcal bacteremia following urgent orotracheal intubation in critically ill patients. AB - OBJECTIVES: To examine whether urgent orotracheal intubation (OI) can induce bacteremia. To find predictive factors for post-intubation bacteremia. DESIGN: Prospective observational study. SETTING: Seventeen-bed medical intensive care unit in a university hospital. PATIENTS: Sixty-eight adult intensive care patients undergoing urgent OI. MEASUREMENTS AND RESULTS: Patients in need of OI could be included if no cardiopulmonary resuscitation was performed. A blood culture was taken immediately before, as soon as possible after, and 60 min after intubation. The indication for intubation, ease of intubation, and the antibiotics used before intubation were registered. Six patients (6/68 or 9%) had streptococcal bacteremia immediately (mean 10.8 min) after intubation. No patient (0/62) had streptococcal bacteremia 60 min after intubation (P = 0.01). Four of the six patients showing streptococcal bacteremia after intubation were intubated by a second doctor because of difficulties during intubation, whereas this was the case in only 9/62 in those without streptococcal bacteremia (P = 0.01). Four of the 13 patients (31%) who needed to be intubated by a second doctor showed transient streptococcal bacteremia. Of the 20 patients not receiving antibiotics at the time of intubation, four (20%) had streptococcal bacteremia compared with 2/47 (4.2%) patients receiving antibiotics (P = 0.06). CONCLUSIONS: Urgent intubation can cause transient bacteremia with streptococci in a significant proportion of intensive care patients. The observed frequency of bacteremia is higher than previously reported after elective intubation. The difficulty of intubation is probably a predisposing factor. PMID- 11396291 TI - Role of thrombolysis in cardiac arrest. PMID- 11396292 TI - Toxic shock syndrome complicating influenza A infection: a two-case report with one case of bacteremia and endocarditis. PMID- 11396293 TI - Usefulness of computed tomography for early detection of pneumonia in leukopenic patients. PMID- 11396294 TI - Surgical management of fulminant pseudomembranous colitis. PMID- 11396295 TI - Prolapsing left atrial myxoma, pulmonary edema and cerebral ischemia. PMID- 11396296 TI - Elevation of prostatic markers following cardiogenic shock. PMID- 11396297 TI - Multiple organ dysfunction dramatically improving with the infusion of vitamin C: more support for the persistence of scurvy in our "welfare" society. PMID- 11396298 TI - Electric cardioversion of atrial flutter in a critically ill patient in the prone position. PMID- 11396299 TI - Stridor and trismus: safe airway by oral awake emergency intubation. PMID- 11396300 TI - Ventilator-induced lung injury. PMID- 11396301 TI - Late steroid therapy in primary acute lung injury. PMID- 11396302 TI - Differential effects of activity and climate on onset of subarachnoid hemorrhage. AB - Conflicting findings of the effect of climate on onset of subarachnoid hemorrhage (SAH) may result from the influence of strenuous activities which can trigger aneurysmal rupture independent of climatological factors. The effect of climate and patient activities on onset of SAH were analyzed. The clinical records of 786 consecutive patients with aneurysmal SAH admitted to our hospital for 10 years were reviewed. Activities at onset were categorized according to the intensity of strain at onset. Seasonal variation, circannual cyclic trend, and association with 90 meteorological factors were examined in each category and the results were compared between categories. Bimonthly occurrence in the light strain group showed a significant seasonal variation and cyclic trend with two peaks in early spring and fall, whereas no significant trend was detected in the overall patients and in the heavy strain group. The significant meteorological factors were global solar radiation, sunshine hours, changes in mean and minimum temperature and mean vapor pressure from the previous day, and minimum pressure in the previous 7 days. Lower global solar radiation in the light strain group was associated with onset with the lowest p value (p = 0.0046). No factors were significant in the heavy strain group. There is some evidence of the possible influence of climatological factors on onset of SAH without strenuous activity. Strenuous activity seems to affect onset more strongly, which masks any effect of climate. PMID- 11396303 TI - Inhibitory effects of verapamil and nitroglycerin on contraction and cytosolic Ca2+ levels in cerebrovascular smooth muscle during chronic cerebral vasospasm. AB - The mechanisms of the inhibitory effects of verapamil and nitroglycerin on vasospasm were investigated by measuring cytosolic Ca2+ level ([Ca2+]i) and muscle tension in 28 normal specimens and 28 spastic vascular specimens of smooth muscle. Experimental vasospasm was produced by the two-hemorrhage method in the canine basilar artery. [Ca2+]i and tension were recorded simultaneously with a fluorimeter using fura-2. High K+ concentration (72.4 mM) and U-46619 (thromboxane A2 analogue, 10(-8) M) were used as stimulants, and the inhibitory effects of verapamil or nitroglycerin on muscle contraction and increased [Ca2+]i were examined. Verapamil inhibited [Ca2+]i and contraction in high K+ concentration-stimulated arteries. Verapamil inhibited [Ca2+]i more strongly than contraction in U-46619-stimulated arteries. There were no significant differences in the effects of verapamil in the control and vasospasm groups. Nitroglycerin inhibited contraction with little effect on [Ca2+]i in high K+ concentration stimulated arteries in both the control and vasospasm groups. Nitroglycerin inhibited contraction with little effect on [Ca2+]i in U-46619-stimulated arteries and the inhibitory effect was weaker in the vasospasm group than in the control group. The inhibitory effects of verapamil on muscle tension and [Ca2+]i in vasospastic vessels were as strong as those in normal vessels. In contrast, the inhibitory effects of nitroglycerin were reduced in vasospastic vessels. Increased Ca2+ sensitivity in vasospastic vessels may have reduced the inhibitory effects of nitroglycerin. PMID- 11396304 TI - Surgical treatment of hemangioblastomas with presurgical endovascular embolization. AB - The usefulness of presurgical embolization of hemangioblastomas was evaluated retrospectively in eight of 30 operations performed on eight of 27 consecutive patients with histologically verified hemangioblastoma. All tumors had over 3.5 cm maximum diameter of the solid mass. Presurgical embolization achieved 100% embolization in one case, 95% to 80% in two, 70% to 50% in two, and less than 50% in three. Total obliteration was obtained only in one case with a single feeding artery. No permanent neurological deficits developed after embolization, but cerebellar infarction occurred in one patient. The surgery was definitely easier than expected in three cases with 80% or more obliteration of tumors. Tumor swelling and cerebellar hematoma occurred during operation in one case with 70% tumor embolization and another case with less than 50%. Blood transfusion during operation was carried out in two cases with less than 50% tumor embolization. The clinical outcome was good recovery in one case, moderate disability in five, and severe disability in two. The reasons for residual neurological deficits were operation and meningitis in one patient, operative and preoperative symptoms in two, and residual preoperative symptoms in four. Neurological deterioration after surgery occurred in three patients with tumor embolization of less than 50%. Partial embolization of hemangioblastomas does not reduce operative complications or morbidity, unless almost complete embolization is achieved, which is not so easy. PMID- 11396305 TI - Participation of thrombospondin-1 in the activation of latent transforming growth factor-beta in malignant glioma cells. AB - Malignant glioma cells secrete transforming growth factor-beta (TGF-beta) and can activate latent TGF-beta. However, the mechanism of the latent TGF-beta activation has not yet been determined. This study examined whether thrombospondin-1 (TSP-1) secreted by malignant glioma cell lines participates in the activation of latent TGF-beta secreted by the glioma cells. Western blot analysis revealed that TSP-1 was present in both the cell lysates and the culture supernatants of all three malignant glioma cell lines (T98G, A172, and U251). A bioassay for TGF-beta activity revealed that all malignant glioma cell lines used in this study could activate latent TGF-beta by themselves. Latent TGF-beta 1 activation, evaluated by enzyme-linked immunosorbent assay, was inhibited by more than 50% by the addition of neutralizing anti-TSP-1 monoclonal antibody or anti TSP-1 polyclonal antibody. These results indicate that TSP-1 has a predominant role in the activation of latent TGF-beta in malignant glioma cells. PMID- 11396306 TI - Ruptured intracranial aneurysm associated with unruptured abdominal aortic aneurysm--case report. AB - A 61-year-old male with hypertension presented with sudden onset of headache and nausea due to subarachnoid hemorrhage (SAH). He had two siblings with history of SAH due to ruptured intracranial aneurysms. Right carotid angiography on admission showed an anterior communicating artery aneurysm. At that time, the extracranial arteries were not examined. The aneurysm was clipped with no complications. A pulsating mass was palpable in the abdomen 37 days after the onset. Ultrasonography and computed tomography showed an abdominal aortic aneurysm with intraluminal thrombus, measuring 8 x 9 x 8 cm. Normal pressure hydrocephalus had already developed. The patient underwent elective abdominal aortic aneurysm resection before ventriculoperitoneal shunting. After shunting, he recovered fully. The present case indicates that unpredictable sudden enlargement of associated abdominal aortic aneurysm is possible in patients with ruptured intracranial aneurysms. PMID- 11396307 TI - "Early" apoplexy due to traumatic intracranial aneurysm--case report. AB - A 76-year-old man presented with a traumatic aneurysm of the left internal carotid artery which caused repeated subarachnoid hemorrhages within 20 hours of a fall from a height. Early computed tomography (CT) detected no brain abnormalities, but repeat CT found subarachnoid hemorrhage. Internal carotid angiography detected a pseudoaneurysm, which was not treated because of his poor clinical condition. He died of multiple organ failure. Early detection of a traumatic intracranial aneurysm is important for the prevention of aneurysmal rupture, or "delayed" apoplexy. Review of 171 cases with traumatic aneurysms from the literature found that false negative angiography occurred only in three cases on post-trauma day 7 and thereafter. Early diagnostic angiography within a week of the initial trauma is indicated if traumatic aneurysm is suspected to detect early signs of irregularity, spasm, and narrowing of the arterial wall. Repeat angiography is indicated if aneurysmal formation is still highly suspected in spite of negative initial angiography. PMID- 11396308 TI - Dissecting aneurysms at the A1 segment of the anterior cerebral artery--two case reports. AB - Two rare cases of dissections which involve the anterior cerebral artery (ACA) are reported. A 58-year-old woman presented with a ruptured dissecting aneurysm manifesting as sudden onset of severe headache and consciousness disturbance followed by aphasia, right hemiparesis, paresis of the left lower extremity, and choreoathetotic movements of the upper arms and face. Computed tomography and angiography revealed subarachnoid hemorrhage due to a dissecting aneurysm at the left A1 segment. The dissecting aneurysm was trapped surgically on the day of onset. Her neurological deficits disappeared within a month. A 39-year-old woman experienced continuous dull headache from the day before onset, and then suffered right hemiparesis. Magnetic resonance (MR) imaging revealed cerebral infarction at the left globus pallidus. Angiography and MR imaging revealed a dissecting aneurysm at the left A1 segment and occlusion of the left Heubner's artery. She received conservative treatment and her neurological findings were improved. Dissections or dissecting aneurysms involving the ACA can be classified into three types: Extension of a dissection to the ACA from the internal carotid artery, dissection at the A1 segment, and dissection at the A2-A4 segments. These types of dissection have distinct uniform clinical features. PMID- 11396309 TI - Neuroradiological features of intraosseous cavernous hemangioma--case report. AB - A 40-year-old man presented to our hospital because of a painless lump on his right forehead. Radiography showed a radiolucent defect in the frontal bone. Bone window computed tomography demonstrated a lucent mass which expanded externally from the diploe destroying and passing through the outer plate. T1-weighted magnetic resonance imaging revealed the lesion as non-homogeneously isointense, and T2-weighted imaging as non-homogeneously hyperintense. The lesion was enhanced non-homogeneously after contrast administration. The lesion was subtotally removed. Histological examination suggested cavernous hemangioma. Preoperative examinations could not provide a definitive diagnosis, which was established by the operative finding. Because of the difficulty of diagnosis in the early stage, surgical treatment in the early stage is recommended for total removal and definitive diagnosis of intraosseous tumor. PMID- 11396310 TI - Large cholesterol granuloma arising from the frontal sinus--case report. AB - A 59-year-old male presented with a large cholesterol granuloma arising from the frontal sinus manifesting as a large, fluctuated, soft mass in his brow, compressing left eye. Skull radiography showed dilation of the frontal sinus. Computed tomography and magnetic resonance imaging revealed a cystic mass extending into the left orbit and anterior cranial fossa. Gross inspection at the frontal craniotomy showed mucinous, dark green fluid intermingled with shining material. The histological diagnosis was cholesterol granuloma with epithelial lining. Large cholesterol granuloma with facial deformity is always associated with bone and cosmetic problems. Wide opening of the frontal sinus followed by cyst wall removal and plastic repair of the skull is necessary. PMID- 11396311 TI - Sacral meningeal cyst associated with valve-like mechanism--case report. AB - A 58-year-old woman presented with low back pain radiating to the lower extremities. Magnetic resonance imaging revealed a cystic lesion in the sacrum compressing the nerve roots. At operation, a valve-like communication was found between the subarachnoid space and the cyst cavity in the vicinity of the sacral nerve root. The communication was obliterated with a purse-string suture and reinforced with a free muscle graft. Postoperatively, she reported improvement of the pain. Valve-like communication between the cyst cavity and subarachnoid space can cause enlargement of spinal meningeal cyst, and could also explain enlargement of sacral meningeal cyst. Surgical obliteration of the communication rather than the cyst resection is more important for sacral meningeal cyst. PMID- 11396313 TI - International effort on geriatric oral health. PMID- 11396312 TI - Palliative care education and training during residency: a survey among residents at a tertiary care hospital. AB - BACKGROUND: Palliative care is the active total care of patients in advanced and incurable stages of cancer. More than 70% of all cancer patients in India require palliative care for relief of pain, other symptoms and psychosocial distress. The need for education and training in palliative care has been emphasized by the World Health Organization (WHO) during the past 15 years. This survey aimed to assess the awareness, clinical knowledge, and education and training aspects of palliative care among the clinical residents of a tertiary care hospital. METHODS: During January and February 1999, a total of 100 residents were asked to respond to a questionnaire on palliative care education and training during residency. The questionnaire consisted of 10 questions (each question with 3 different responses). Awareness regarding palliative care was assessed by 3 questions; clinical knowledge by 4 questions; and education and training obtained during residency by 3 questions. Comparisons were made using the Chi-square test between the oncology and non-oncology resident groups. RESULTS: Forty-nine residents (23 oncology and 26 non-oncology) responded to the survey. Awareness that palliative care involves active total care was lacking in 8 (16%) residents. The cost of palliative care in India was considered high by 17 (65%) non-oncology residents and 21 (43%) of the whole group. Hospice, as the right place for palliative care, was chosen by 14 (61%) oncology and 3 (11.5%) non-oncology residents (p = 0.0003). In the whole group, 21 (43%) believed that palliative care could be provided on an outpatient basis. Pain, depression and cachexia were identified as the most distressing symptoms of patients with incurable cancer. Seventeen (83%) oncology and 4 (15%) non-oncology residents (p = 0.000) knew how to use the WHO step-ladder for cancer pain relief. All oncology residents and 10 (38%) non-oncology residents preferred the oral route for providing medications for pain relief (p = 0.0001). The didactic education and training imparted during residency was considered as 'not enough' by 39% of oncology and 62% of non oncology residents (51% overall). The confidence to deliver quality palliative care was lacking in 43% and 58% of oncology and non-oncology residents, respectively. CONCLUSION: Clinicians in India need to be provided focused skills and training for them to be able deliver quality palliative care to the large number of patients with incurable cancer. The cost of palliative care and the optimum place to deliver it, the symptoms of advanced cancer, pain relief and symptom control methods and quality of life in end-stage cancer patients are some aspects that should be an integral part of clinical residency programmes. PMID- 11396314 TI - Searching Medline effectively. PMID- 11396315 TI - Newborn health agenda: do we need another worker? PMID- 11396316 TI - Spectrum of clinical manifestations in severe falciparum malaria in Koraput district, Orissa. PMID- 11396317 TI - Nursing personnel and prevention of nosocomial infections. PMID- 11396318 TI - Kerala pesticide tragedy. PMID- 11396319 TI - The challenges of palliative care in India. PMID- 11396320 TI - A People's Charter for Health and beyond. PMID- 11396321 TI - Radioiodine kinetics and thyroid function following the universal salt iodization policy. AB - BACKGROUND: Universal salt iodization was introduced in Delhi in 1989. The present study quantifies the change in iodine kinetics as a result of this. The previous values were reported 10-30 years earlier, when Delhi was iodine deficient. METHODS: Thirty subjects (18 men and 12 women, 17-48 years of age) who were residents of Delhi and had no thyroid disorder, were recruited from our outpatient clinic in 1999. The 24-hour urinary excretion of iodine and the iodine content of salt consumed at home by these subjects were estimated. Kinetic studies of iodine using radiotracer 131I were done to determine thyroid iodine clearance, renal iodine clearance, percentage uptake and absolute iodine uptake by the thyroid gland, and plasma inorganic iodine. RESULTS: The median 24-hour urinary iodine excretion was 341.3 micrograms. The mean (SD) thyroid uptake of radioactive iodine was 4.9 (2.3)% at 2 hours and 19.1 (8.0)% at 24 hours. The median calculated plasma inorganic iodine was 1.36 micrograms/dl, absolute iodine intake 6.5 micrograms/hour and thyroid iodine clearance was 4.8 ml/minute (geometric means 1.68 micrograms/dl, 8.5 micrograms/hour and 8.1 ml/minute, respectively). The serum thyroid hormones and thyroid stimulating hormone were within normal limits. CONCLUSION: Compared to the values reported 10-30 years ago when the population was iodine deficient, the present urinary iodine excretion, plasma inorganic iodine and absolute iodine intake have increased, while the percentage thyroid uptake of iodine ingested and thyroid clearance have decreased. The lack of change in the serum thyroid hormone levels after 10 years of universal salt iodization indicates that iodine consumption has had no adverse effect on thyroid function in these normal individuals. These changes are consistent with the increase in iodine consumption. Since the iodine ingestion in a community may change with time, assessment of iodine kinetics should be done periodically in different regions of the country. PMID- 11396322 TI - Immunosuppression in live-related donor renal transplantation. AB - BACKGROUND: Triple immunosuppression with cyclosporine, azathioprine and prednisolone is the most common regimen employed following renal transplantation. No information is available regarding its impact on the results of renal transplantation in India. The present study is an audit of a fixed-dose cyclosporine-based immunosuppressive regimen in an exclusively live-related donor transplant programme, with specific regard to graft and patient outcomes. METHODS: Patients transplanted over a 3-year period and receiving cyclosporine based immunosuppression were studied. The relationship between immunosuppression and graft outcomes [rejection episodes (RE), graft function, graft survival], and patient outcomes (patient survival) was analysed in those receiving triple immunosuppression. Dosage schedules were audited. Cyclosporine trough level monitoring was employed at graft dysfunction episodes, or at dose reduction points. RESULTS: The median follow up was 14 months. Triple drug immunosuppression was used in 191 patients and double drug therapy in 26. The overall one-year patient survival rate was 91% and the corresponding graft survival rate was 90%. An audit of dosing schedules showed that over the first 6 months post-transplant, cumulatively, 20%-50% of patients received azathioprine, and 55%-60% received cyclosporine in doses below the protocol. The immunosuppressive doses (both of cyclosporine and azathioprine) in the first month were significantly related to the RE (p < 0.01) in the first month and the total number of RE in the first 6 months (p < 0.01). The other predictors were younger recipient age and older donor age. The sixth-month serum creatinine level was predicted by the donor age, the level of serum creatinine in the first month and the total number of RE in the first 6 months post-transplant. While no specific predictors of graft loss were identified in this cohort, diabetic nephropathy (p = 0.000) as the native renal disease, and the total number of RE were strongly related to patient mortality. The occurrence of > or = 2 RE in the first 6 months was an independent predictor, increasing the risk of death in the first 2 years post-transplant by 2.3 (p = 0.0001, 95% CI: 1.5-3.4). CONCLUSIONS: Sub-therapeutic baseline immunosuppression in the early post-transplant period predisposes to acute RE. This has an impact not only on graft function but also forms an important proximate marker of mortality, as seen in this cohort. Thus, immunosuppressive drug dosage should be optimized and therapeutic drug level monitoring strategies should be preemptive rather than event related, especially in the early post-transplant period. While fixed-dose immunosuppressive drug schedules are widely followed, it is possible to fall short of the target unless a specific effort is made to meet and sustain schedules. PMID- 11396323 TI - Carrier detection and prenatal diagnosis in families with haemophilia. AB - BACKGROUND: Haemophilias are the commonest X-linked disorders affecting approximately 1 in 10,000 male births. Detection of carrier women in families with haemophilia and subsequent antenatal diagnosis of confirmed carriers are important services for these patients and their relatives. Over the last 6 years we performed carrier detection and antenatal diagnosis in families with patients of haemophilia A and B. METHODS: During the last 6 years, 159 families with haemophilia A and B were analysed for carrier detection by DNA analysis, using various polymorphic markers of factors VIII and IX genes. The polymorphisms used were intron 18 Bcl I, intron 19 Hind III, intron 22 Xbal and DXS52/St14 of the factor VIII gene and intron I Ddel, intron 4 Taql, 3 Hhal and Residue 148 codon Mnll of the factor IX gene. There were 189 probable carriers (whose carrier status was not known) and 99 obligatory carriers (confirmed carriers by family pedigree analysis) from 102 families with haemophilia A. Of the 57 families with haemophilia B analysed, there were 98 probable and 52 obligatory carriers. All the analyses were carried out by polymerase chain reaction. For antenatal diagnosis, prior to polymorphism analysis, the sex of the foetus was detected by Y chromosome-specific amplification. RESULTS: One hundred and four females were diagnosed as carriers and 63 as non-carriers by the intragenic polymorphic markers in families with haemophilia A. Eighteen women were informative with only the extragenic marker of factor VIII gene. Four women were not informative with any of the markers used. In families with haemophilia B, 37 women were diagnosed as carriers and 34 as non-carriers by the intragenic markers and 34 were informative only with the extragenic markers. Seventeen women were not informative with any of the markers used. Of the 25 antenatal diagnoses performed (20 haemophilia A, 5 haemophilia B) using the same markers as those used in carrier detection, 14 were male foetuses and 11 female as detected by Y chromosome-specific polymerase chain reaction. Eight were affected males and 6 unaffected. Among the females, 5 were carriers and 6 normal. CONCLUSION: Using the above polymorphic markers of factors VIII and IX genes, a diagnosis could be made in the majority of families. PMID- 11396324 TI - Hepatopulmonary syndrome: clinical perspectives. AB - Hepatopulmonary syndrome consists of a triad of chronic liver disease, pulmonary gas exchange abnormalities and pulmonary vascular dilatation in the absence of detectable cardiopulmonary disease. Patients usually present with symptoms of liver disease and the clinical recognition of this condition is a challenge. Newer non-invasive tests facilitate the diagnosis. Therapeutic strategies for this condition are still dismal. Liver transplantation is a possible curative option for a subgroup of patients with hepatopulmonary syndrome. PMID- 11396325 TI - The death knell for dopamine? PMID- 11396326 TI - Enthesopathy: clinical recognition and significance. AB - Enthesopathy is a common clinical finding denoting pathology at the 'entheses', i.e. attachment sites of muscles, tendons, joint capsules, ligaments and fascia to the bone. Inflammatory enthesopathy or enthesitis is a sine qua non of seronegative spondyloarthropathies (SSA). It can also be occupational, metabolic, drug induced, infective or degenerative. Bursitis closely mimics enthesitis. Ultrasound with high frequency transducers is a simple, cost-effective and feasible test to detect enthesopathy which is amenable to treatment with local steroid injections, physiotherapy and non-steroidal anti-inflammatory drugs, in addition to treatment of the primary disease. Unrecognized and untreated, it can lead to considerable morbidity. PMID- 11396327 TI - Nucleic acids in diagnosis (Part II): Genetic testing and screening. PMID- 11396329 TI - Kibbles and bytes. PMID- 11396328 TI - Empowered opinion leaders in India and the HIV/AIDS programme. PMID- 11396331 TI - Galactic archaeology. PMID- 11396330 TI - A touch of poison. PMID- 11396332 TI - Robotic bombers. PMID- 11396333 TI - New trick from old dog. PMID- 11396334 TI - Save the earth. PMID- 11396335 TI - Unfair game. PMID- 11396336 TI - The American terrorist. PMID- 11396337 TI - The mice that warred. PMID- 11396338 TI - A license for copycats? PMID- 11396339 TI - Fox's flapdoodle. PMID- 11396340 TI - Piloting through uncharted seas. PMID- 11396341 TI - The paradox of the sun's hot corona. PMID- 11396342 TI - Solving the mystery of insect flight. PMID- 11396343 TI - Sign language in the brain. PMID- 11396344 TI - North to Mars! PMID- 11396345 TI - Hair. Why it grows why it stops. PMID- 11396346 TI - The Himba and the dam. PMID- 11396347 TI - A low--pollution engine solution. PMID- 11396348 TI - [Interactions of food and drug metabolism]. AB - The nutritional state, and/or the ingestion of specific nutrients, is/are able to modify drug disposition, by interfering with drug absorption, distribution, storage, and metabolism. Recent data report that nutrients interfere with drug metabolism either by modifying key enzymes of phase I (cytochromeP450 dependent mixed function oxidase) and II (glucuronosyl, sulfonyl- ... transferases), or by modulating coenzymes availability (NADPH, UDPglucuronic acid...). Food components involved in drug metabolism modifications are either macro-nutrients (carbohydrates, lipids, proteins, ethanol), micronutriments (vitamins, minerals), or phytochemicals. Drug-nutrients interactions may be beneficials, and thus could constitute, i.e. a way to improve drug therapeutic index, or generate adverse effects. PMID- 11396349 TI - [Nutrition and digestive disorders in children]. PMID- 11396350 TI - [Functional foods: concepts and strategy]. PMID- 11396351 TI - [Pharmacological treatment of obesity and syndrome X]. PMID- 11396352 TI - [Volatile N-nitrosamines in meat]. PMID- 11396353 TI - Evidence for cure of 'young' men with prostate cancer. AB - A study was undertaken to evaluate the question of cure in "young" men with prostate cancer treated by external-beam radiation. Results in young men (< or = 65 years) were compared to older men. Biochemical freedom from failure was examined to 10 years' follow-up, and hazard functions for failure vs time were reported. Results show that prostate cancer patients are cured by external-beam radiation and that there is no difference in results for young or older men. Few failures occur after 5 years' follow-up and the percentage cured is similar to that with prostatectomy, with much less morbidity. Appropriate dose is necessary to optimize outcome. PMID- 11396354 TI - Gynecologic malignancies in older women. AB - The aging of the population is a social phenomenon that will present a challenge to clinical practice in the 21st century. Women constitute a majority of the elderly population as they outlive males by 5 to 7 years. Ovarian, endometrial, and vulvar cancers are diseases seen more commonly in postmenopausal and elderly women. Cervical cancer continues to be a significant problem in the elderly and is usually detected at a later stage in that population than in younger patients. Accordingly, primary care clinicians ought to possess a thorough knowledge of gynecologic malignancies and should refer women who present with these disorders to a gynecologic oncologist. Ovarian cancer patients treated by a gynecologic oncologist are more likely to undergo proper surgical staging, leading to optimal debulking surgery and improved survival. Age, by itself, should not alter the diagnostic and therapeutic approach to gynecologic malignancy. Elderly patients can safely undergo radical pelvic surgery. Multiagent chemotherapy is also possible in the elderly without excess morbidity, and without compromise of response rates. Radiation therapy for cervical cancer appears to be as effective and is generally well tolerated. The Papanicolaou (Pap) test continues to be the primary screening tool for cervical cancer. Although transvaginal ultrasound seems to be useful in detecting early-stage ovarian cancer, its cost effectiveness for screening the general population remains to be demonstrated. The main considerations in the treatment of ovarian, endometrial, cervical, and vulvar cancer are discussed. PMID- 11396355 TI - Clinical trials referral resource. Health-related quality of life in cancer clinical trials. PMID- 11396356 TI - Cognitive function after systemic therapy for breast cancer. AB - An underinvestigated area of breast cancer survivorship involves the possible impairment of cognitive function following adjuvant chemohormonal therapy. Numerous reports of disturbing and disruptive changes in short- and long-term memory, attention span, concentration, and language skills have been made by breast cancer patients who have received chemotherapy. This article reviews the four published studies that have documented cognitive dysfunction following adjuvant chemohormonal therapies commonly used in breast cancer. The studies describe a subset of approximately one-third of participants who experienced long term cognitive impairment. Patient- and treatment-related factors that may influence cognitive function are outlined. The impact of these cognitive impairments on the individual breast cancer survivor's quality of life is discussed, as is the potential overall impact of this research on future adjuvant therapy. The need for a prospective longitudinal study documenting the neuropsychological sequelae of adjuvant chemohormonal therapy is emphasized. PMID- 11396357 TI - Incidence and management of AIDS-related lymphoma. AB - Over time, the spectrum of the acquired immune deficiency syndrome (AIDS) epidemic has changed, especially with the advent of highly active antiretroviral therapy (HAART). The goal of this article is to delineate changes occurring in the incidence and management of lymphoma over the course of the AIDS epidemic. Lymphoma usually occurs rather late in the course of human immunodeficiency virus (HIV) infection and is the cause of death in up to 20% of HIV-infected individuals. It is seen in all population groups at risk for HIV and is more common in men than in women. It is usually diagnosed in patients with markedly decreased CD4 cell counts, consistent with prolonged periods of HIV infection and subsequent immunosuppression. Recent data from several large series have demonstrated a substantial decline in the median CD4 cell count among patients with newly diagnosed AIDS-related lymphoma despite the recent widespread use of HAART. While still somewhat controversial, use of HAART has generally not produced a significant decline in the incidence of AIDS-related lymphoma. Patients treated with low-dose vs standard-dose chemotherapy for AIDS-related lymphoma have achieved similar response and survival rates, although standard dose therapy is associated with greater toxicity. Adapting therapy to prognostic factors has not produced a significant improvement in survival. Use of antiretroviral therapy along with chemotherapy appears safe, and may be associated with longer survival. An infusional regimen called EPOCH (etoposide, prednisone, vincristine [Oncovin], cyclophosphamide, doxorubicin HCl) shows promise in the future management of AIDS-related lymphoma. No regimen is currently considered the standard of therapy for patients with relapsed AIDS related lymphoma, and survival is short in this setting. PMID- 11396358 TI - Transplant registries: guiding clinical decisions and improving outcomes. AB - About 50,000 hematopoietic stem cell transplantations are performed yearly, primarily for malignancies. Use of this therapy increased dramatically over the past 30 years due to its proven and potential efficacy in diverse diseases, better understanding of appropriate timing of transplantation and patient selection, and greater availability of allogeneic donors. The International Bone Marrow Transplant Registry (IBMTR) and the Autologous Blood and Marrow Transplant Registry (ABMTR) collect data on consecutive allogeneic and autologous transplants, respectively, in more than 400 participating centers worldwide. The IBMTR/ABMTR database contains information on more than 120,000 transplant recipients. Among 11,347 patients transplanted in 101 IBMTR/ABMTR research centers in North America during 1995-1997, 66% received autologous transplants, 24% related-donor transplants, and 10% unrelated-donor transplants. More than 90% of transplantations were for malignant disease, with more than half of these done in patients with advanced disease. Of the recipients, 70% were younger than 50 years. Posttransplant survivals varied substantially by disease, transplant type, recipient age, and disease status at transplantation. IBMTR/ABMTR data provide an important tool for assessing transplant use and outcome, identifying prognostic factors for transplant outcomes, evaluating new transplant therapies, comparing transplant and nontransplant therapies, evaluating late transplant complications, and planning prospective phase II and III clinical trials. PMID- 11396359 TI - Dose-dense and sequential strategies in adjuvant breast cancer therapy. AB - Several attempts have been made to improve the survival rates of breast cancer patients. The benefit of adjuvant chemotherapy was clearly shown, but the absolute difference of 2% to 11% in overall survival, depending on the patient group, is disappointingly small. In particular, high-risk patients, such as those with > or = 10 involved lymph nodes, extracapsular spread, or vascular invasion, still have an excessive risk of recurrence even after standard adjuvant chemotherapy. To increase the survival rates after adjuvant therapy, new chemotherapeutic agents and new strategies of application are currently being evaluated in clinical trials. Chemotherapy with cyclophosphamide (Cytoxan, Neosar), methotrexate, and fluorouracil (CMF) seems to be safe and effective in patients with breast cancer. In addition, in metastatic patients, dose intensified chemotherapy is being investigated. The introduction of epirubicin (Ellence), an agent less cardiotoxic and equally active compared to doxorubicin, enabled the escalation of anthracyclines in adjuvant therapy without serious cardiotoxic effects. The combination of dose-intensified chemotherapy and sequential application in the treatment of breast cancer is reviewed. PMID- 11396360 TI - Epirubicin/taxane combinations in breast cancer: experience from several Italian trials. AB - Doxorubicin/paclitaxel (Taxol) combinations are very active in advanced breast cancer, with objective response rates up to 90%, but have shown a high incidence of cardiotoxicity. A phase I/II trial replacing doxorubicin with epirubicin (Ellence), a less cardiotoxic analog, produced an objective response rate of 84%, but with a low rate of cardiotoxicity. A careful cardiac monitoring in more than 100 patients treated with this combination has demonstrated that the risk of congestive heart failure is below 10% up to a cumulative epirubicin dose of 990 mg/m2. To examine the possibility that the pharmacokinetic and pharmacodynamic interactions that occur when anthracycline and paclitaxel are administered together might result in subadditive antitumor activity, a phase III study is comparing concomitant vs sequential administration of epirubicin and paclitaxel in patients with advanced breast cancer. A phase I/II study of epirubicin plus docetaxel as first-line chemotherapy for advanced breast cancer patients evaluated the maximum tolerated doses and for subsequent studies recommended epirubicin at 75 mg/m2 plus docetaxel at 80 mg/m2. In the adjuvant setting, an ongoing phase III trial is comparing epirubicin plus paclitaxel vs FEC (fluorouracil, epirubicin, and cyclophosphamide [Cytoxan, Neosar]) in node positive patients. Preliminary data confirm the cardiac safety of these treatments. PMID- 11396361 TI - New combinations with epirubicin in advanced breast cancer. AB - Several trials have shown that anthracyclines and taxanes can be combined to achieve response rates ranging from 70% to 90%, with complete responses ranging from 19% to 41%. In an attempt to increase the activity while maintaining tolerability, gemcitabine (Gemzar) was added to the epirubicin (Ellence)/paclitaxel (Taxol) regimen. Among 36 metastatic breast cancer patients treated with this new combination, the overall response rate was 92%, including 31% with a complete response. Another attempt to improve the outcome of metastatic breast cancer patients involves a phase III multicentric randomized trial (MANTA-1) to evaluate if paclitaxel maintenance therapy after anthracycline/taxane combination therapy can improve time to progression and overall survival. Although anthracyclines are more frequently used in the adjuvant setting, it is important for the clinicians to know whether this class of drugs can be used again for those patients who develop metastatic disease. An analysis of 312 patients treated with epirubicin containing regimens as first line treatment for metastatic disease shows that epirubicin-based regimens are active in patients already exposed to anthracyclines in the adjuvant setting, and that the risk of cardiac toxicity is low up to a cumulative epirubicin dose of 990 mg/m2. PMID- 11396362 TI - Aromatase inhibition and antiestrogen therapy in early breast cancer treatment and chemoprevention. AB - The aromatase inhibitors represent an important class of hormonal agents for the management of breast cancer. The third-generation aromatase inhibitors have replaced megestrol acetate as second-line hormonal therapy in advanced breast cancer, and large clinical trials are maturing to establish their efficacy relative to tamoxifen (Nolvadex) in the first-line metastatic setting. The increased potency, increased specificity, and established efficacy of aromatase inhibitors in advanced breast cancer have provided the rationale for a large number of randomized trials in the adjuvant setting evaluating anastrozole (Arimidex), exemestane (Aromasin), and letrozole (Femara). These trials are addressing the value of these agents in sequence with, instead of, and in combination with tamoxifen. The relationship between estrogen exposure and breast cancer risk has long been accepted and traditionally related to estrogen-receptor mediated events. The emergence of the estrogen genotoxicity hypothesis as a mechanism for breast cancer carcinogenesis provides additional rationale for considering aromatase inhibitors in the chemoprevention setting. PMID- 11396363 TI - Adjuvant exemestane therapy after 5 years of tamoxifen: rationale for the NSABP B 33 trial. AB - Tamoxifen (Nolvadex) has long been established as "standard" adjuvant therapy for receptor-positive, early-stage breast cancer. Results from clinical trials suggest that after approximately 5 years, tamoxifen may lose its effectiveness and may even become harmful if not stopped. At the time of tamoxifen discontinuation, "seemingly" disease-free patients may still have residual micrometastatic tumor cells. In a proportion of such patients, these cells may still be responsive to tamoxifen and thus could grow as a result of stopping the drug. The majority of clinical information, however, suggests that by 5 years of therapy, a greater proportion of patients will have micrometastatic tumor cells that have become resistant to tamoxifen and will be stimulated by continuation of the drug for a longer time. Because in both such cases the micrometastases are hormonally sensitive, a reasonable approach--in addition to stopping tamoxifen- is to decrease the level of estrogenic stimulation by introducing an aromatase inhibitor. The National Surgical Adjuvant Breast and Bowel Project is launching a clinical trial (NSABP B-33) to evaluate the sequential administration of 2 years of exemestane (Aromasin), a steroidal aromatase inactivator, after 5 years of tamoxifen in postmenopausal, receptor-positive patients who are disease-free at the time of tamoxifen discontinuation. PMID- 11396364 TI - Estramustine potentiates taxane in prostate and refractory breast cancers. AB - Estramustine is nornitrogen mustard linked to estradiol. It binds to tubulin and to microtubule-associated proteins, depolymerizes cytoplasmic microtubules, and disrupts the nuclear matrix. It has limited clinical activity as a single agent, but preclinical studies suggest that it is an effective modulator of antitubulins. This paper reviews the rationale for the combination of estramustine with antitubulins and the clinical toxicity profile of estramustine. Also discussed are data from phase II studies in hormone-resistant prostate cancer and in taxane-resistant breast cancer that suggest that the modulation of antitubulins by estramustine that has been demonstrated in vitro is indeed clinically relevant. Finally, current approaches to improving the tolerability of estramustine are described. PMID- 11396365 TI - Liposomal-encapsulated chemotherapy: preliminary results of a phase I study of a novel liposomal paclitaxel. AB - Liposome encapsulation of antineoplastic drugs entered clinical testing in the late 1980s. As carriers for a variety of agents, liposomes can allow successful delivery of agents that may be subject to rapid degradation in the serum and can modify the toxicity profile. In general, liposomes have demonstrated an ability to attenuate toxicities by their different pharmacokinetic profile and pattern of distribution. Differences in the constitution of the liposome can greatly affect the pharmacokinetic profile resulting in different patterns of toxicity. Characteristics such as size, charge, composition, and integrity can affect performance of the liposome. Liposome encapsulation of doxorubicin has been shown to reduce cardiac toxicity. Preliminary data suggest that encapsulation of paclitaxel can greatly modify neurotoxicity without the need for cremephor. PMID- 11396366 TI - Optimizing adjuvant breast cancer chemotherapy: rationale for the MA.21 study. AB - Recently initiated is a phase III randomized trial (MA.21 trial) of adjuvant chemotherapy for node-positive and high-risk node-negative, premenopausal and postmenopausal (< or = 60 years) women with breast cancer who have no distant metastases. Conducted by the National Cancer Institute of Canada-Clinical Trials Group, the trial will compare two standard therapies, CEF (cyclophosphamide [Cytoxan, Neosar], epirubicin [Ellence], fluorouracil) and AC-->T (doxorubicin [Adriamycin], cyclophosphamide, followed by paclitaxel [Taxol]), and includes a third arm consisting of a dose-dense, dose-intense EC-->T regimen (epirubicin, cyclophosphamide, followed by paclitaxel). These regimens were chosen for study based on results of previous clinical assessments of adjuvant therapies, which, taken together, suggest that CEF, FEC 100 (where 100 represents the dose in mg/m2 of epirubicin in FEC [fluorouracil, epirubicin, cyclophosphamide]), CAF (cyclophosphamide, doxorubicin, fluorouracil), and AC-->T may all be superior to standard AC or CMF (cyclophosphamide, methotrexate, fluorouracil) regimens. This article reviews trial results that support the testing of the regimens chosen for the MA.21 trial. The intent of the MA.21 study is to advance our ability to provide optimal adjuvant therapy for patients with breast cancer. PMID- 11396367 TI - Maternal mortality. A global perspective. AB - Despite difficulty in collecting data and prevailing underreporting and misclassification, data collection by the World Health Organization shows that more than half a million maternal deaths occur globally every year; 99% occur in developing countries and 1% occur in industrialized ones. The maternal mortality rate is highest in sub-Saharan African countries, followed by South Asian countries. Compelling evidence suggests that a reduction in maternal mortality on a short-term basis is possible only by providing modern obstetric care to the 15% of pregnancies that develop complications and making such care available in time. For long-term reduction and prevention of maternal death, preventive measures should be started before conception and should be continued during pregnancy and the postpartum period. PMID- 11396368 TI - Elevated E2: oocyte ratio in women undergoing IVF and tubal ET. Correlation with a decrease in the implantation rate. AB - OBJECTIVE: To investigate the influence of various estradiol (E2): oocyte ratios on reproductive outcome in women undergoing in vitro fertilization and tubal embryo transfer (IVF-TET). STUDY DESIGN: Two hundred seven women undergoing 251 IVF-TET cycles were recruited in this retrospective study. All the women received a flare-up gonadotropin releasing hormone agonist (GnRHa) protocol to achieve ovarian hyperstimulation. Oocyte retrieval was performed 34-36 hours after human chorionic gonadotropin (hCG) injection, followed by TET two days later. RESULTS: An E2: oocyte ratio > or = 350 pg/mL had a higher E2 level (2,213 +/- 2,258 vs. 1,553 +/- 972 pg/mL, P < .05) and fertilization rate (77 +/- 23 vs. 64 +/- 23%, P < .001) but a lower oocyte number (4.8 +/- 4.7 vs. 7.6 +/- 4.8, P < .001) than in those with a ratio < 350 pg/mL. The pregnancy (17.9% vs. 32.8%, P = .03) and implantation (5.3% vs. 12.9%, P = .008) rates were significantly decreased in cycles with an E2: oocyte ratio > or = 350 pg/mL as compared to those with a ratio < 350 pg/mL. CONCLUSION: IVF-TET cycles with an elevated E2: oocyte ratio correlated with lower pregnancy and implantation rates. The poor reproductive outcome possibly was due to the relatively high E2 concentration, which might have a detrimental effect on endometrial receptivity. PMID- 11396369 TI - Endometrial brush biopsy for the diagnosis of endometrial cancer. AB - OBJECTIVE: To evaluate a new technique for processing endometrial cytology for the diagnosis and exclusion of endometrial cancer. STUDY DESIGN: All women at risk for endometrial cancer with clinical indications for endometrial biopsy were evaluated by endometrial brush biopsy (Tao Brush, Cook OB-GYN, Bloomington, Indiana) and Pipelle (Cooper Surgical, Shelton, Connecticut) endometrial biopsies during one office visit. Patients were followed longitudinally for the development of endometrial cancer or until undergoing dilatation and curettage or hysterectomy. All comparisons were analyzed using the chi 2 or t test. RESULTS: One hundred one women (mean age, 58; range, 35-86) had endometrial biopsies performed. Median follow-up was > 21 months (range, 3-29). Twenty-two had cancer or atypia, while the remaining had benign diagnoses. When correlated with the final diagnosis, the Tao Brush had 95.5% sensitivity and the Pipelle, 86% sensitivity. Both devices had 100% specificity, positive predictive value of 100% and negative predictive value of 98%. When the results of the two biopsy devices are considered together, the positive and negative predictive value for detecting or excluding endometrial cancer was 100%. Based on 1998 Medicare reimbursements, a simultaneous second office biopsy using the Tao brush could save approximately $67 per case as compared to a sonohistogram and much more when compared to dilatation and curettage. CONCLUSION: Endometrial cancer can be reliably detected and excluded using these two distinct office biopsy devices simultaneously during one office visit. In patients with an indication for endometrial biopsy, no further diagnostic test may be necessary to exclude or diagnose endometrial cancer or atypia. PMID- 11396370 TI - Effect of transitory hyperprolactinemia on in vitro fertilization of human oocytes. AB - OBJECTIVE: To examine the changes in plasma prolactin (PRL) during ovarian hyperstimulation (OH) and the influence of hyperprolactinemia on folliculogenesis, oocyte retrieval and in vitro fertilization (IVF) success rates and the usefulness of the metoclopramide (MCP) test in predicting the onset of hyperprolactinemia. STUDY DESIGN: Forty-nine cycles of OH were induced in 32 infertile women using follicle-stimulating hormone, human menopausal gonadotropin and human chorionic gonadotropin (GI) (n = 36), also in association with gonadotropin-releasing hormone (GII) (n = 13). The MCP test (10 mg, intravenously) was performed on fertile control women (control group, n = 9) and in GI (n = 21) and GII (n = 8) patients. RESULTS: Plasma PRL and estradiol levels increased during OH, reaching maximum levels on the day preceding oocyte retrieval in GI and GII. Since these two groups exhibited similar PRL curves, they were evaluated as a single group. Patients showing an increase in PRL of > 200% presented a greater number of follicles with a mean diameter > or = 12 mm and more mature oocytes and better IVF success rates than patients with a PRL increase < or = 200%. Oocyte retrieval did not differ between the groups. The MCP test showed hyperresponsiveness in the three groups studied, but no correlation was found between the PRL increase in this test and that during OH. CONCLUSION: Plasma PRL and estradiol levels increase during OH, while the MCP test cannot predict the onset of hyperprolactinemia. Transitory hyperprolactinemia seems to be associated with an increase in the numbers of follicles with a mean diameter > or = 12 mm and with more mature oocytes and better IVF success rates. PMID- 11396371 TI - Effects of unkei-to, an herbal medicine, on endocrine function and ovulation in women with high basal levels of luteinizing hormone secretion. AB - OBJECTIVE: To treat anovulatory women with high plasma LH levels with unkei-to. STUDY DESIGN: A total of 100 patients with polycystic ovary syndrome (n = 38) or non-polycystic ovary syndrome (n = 62) were allocated randomly into two groups. Endocrine levels (FSH, LH and estradiol), follicle growth and response to unkei to were compared for 52 subjects in the unkei-to group and 48 subjects in the control group. RESULTS: Unkei-to induced significant decreases in plasma LH in polycystic ovary syndrome and non-polycystic ovary syndrome with high LH levels. Plasma estradiol levels increased significantly (43.5%) in eight weeks of treatment with unkei-to. Significant development of the dominant follicle was also observed in patients treated with unkei-to. When women suffering from ovulatory failure or irregular menstruation and having high plasma LH concentration were treated with unkei-to for eight weeks, a reduction in the serum LH level of at least 1 SD from baseline was noted in 45.5% of polycystic ovary syndrome patients and in 80.0% of non-polycystic ovary syndrome patients. The mean rate of reduction of serum LH was 22.2 +/- 35.7% in polycystic ovary syndrome patients and 49.7 +/- 15.3% in non-polycystic ovary syndrome patients. This reduction was significant in the non-polycystic ovary syndrome patients (P = .030). The rate of menstrual cycle improvement, including successful ovulation, was 50.0% in the polycystic ovary syndrome group and 60.0% in the non-polycystic ovary syndrome group, with no significant difference between the two groups. PMID- 11396372 TI - Teaching the Laufe-Piper forceps technique at cesarean delivery. AB - OBJECTIVE: To present a method of teaching forceps technique during cesarean delivery of breech-presenting infants using Laufe-Piper forceps and to evaluate its usefulness. STUDY DESIGN: For several years, residents at the University of Texas Medical Branch, Galveston, have learned and practiced Piper forceps technique during cesarean delivery. To assess their experience with this method, we mailed questionnaires to third- and fourth-year residents and recent graduates of the Galveston program. The same surveys were mailed to a control group of residents and recent graduates of two other programs where this teaching exercise is not practiced routinely. RESULTS: Responses were received from 32 (74%) study subjects and 63 (71%) controls. Demographic characteristics and experience with vaginal breech delivery were similar between the two groups. Respondents from the Galveston program noted greater annual use of forceps for vaginal delivery of cephalic-presenting infants (P = .012). They also rated themselves as more comfortable (P = .023) and more skilled (P = .006) with Piper forceps than controls. Of 53 respondents who had had previous experience with this teaching method, 47 noted that it provided a great or moderate educational benefit, and 36 strongly or moderately believed it gave them more confidence in using Piper forceps during vaginal breech delivery. Using multiple regression analysis, sex, overall level of experience, Piper forceps experience during vaginal delivery and overall forceps use were stronger determinants of self-rated comfort and skill than was experience with Laufe-Piper forceps during cesarean. CONCLUSION: Laufe Piper forceps can be used for cesarean delivery of breech-presenting infants. This practice promotes confidence and skill for their use at vaginal delivery. PMID- 11396373 TI - Labor and membrane rupture in twin gestation. Can they affect the ability to estimate fetal weight? AB - OBJECTIVE: To determine whether labor or fetal membrane rupture adversely affects the ability of ultrasound to accurately estimate fetal weight (EFW) in a twin gestation. STUDY DESIGN: A medical record audit was performed of the two-year experience of women delivering live born twins and undergoing a complete ultrasound examination within 72 hours of delivery. Those patients experiencing labor or membrane rupture at the time of the previous ultrasound examination were compared to those without these factors to determine if there was any detrimental impact on the accuracy of ultrasound to EFW. RESULTS: The overall ability of ultrasound to accurately EFW was in the range of 64-76%. This was not significantly different from the accuracy in women experiencing labor (62-80%) or in women with membrane rupture (63-81%). The ability of ultrasound to detect fetal discordance is this series ranged from 78% to 100%, with specificity that ranged from 84% to 89%. CONCLUSION: The ability of sonography to accurately EFW as well as detect discordant twin gestation is not altered by the presence of labor or membrane rupture. PMID- 11396374 TI - Mechanisms of twinning. I. Effect of environmental diversity on genetic expression in monozygotic multifetal pregnancies. AB - OBJECTIVE: To examine, using fingerprint homology, the interaction of genetic and environmental factors in arriving at the final phenotypic expression in uniovular triplets, quadruplets and quintuplets. STUDY DESIGN: The fingerprints of several multifetal sets were collected and matched. A total of 250 fingers were printed. A comparison based on the classic dactylographic patterns was made. Zygosity of each set was known from placental chorionicity studies, as well as blood typing and physical similarities. RESULTS: As the monozygotic set size increased, the ipsilateral print concordance diminished. At one extreme, with twins there is 88% homology; at the other end, with quintuplets the correspondence is 71%. CONCLUSION: The overall trend of these results paralleled increasing intrauterine environmental diversity and tended to support the volar pad pressure hypothesis of prenatal fingerprint development. PMID- 11396375 TI - Mechanisms of twinning. II. Laterality and intercellular bonding in monozygotic twinning. AB - OBJECTIVE: To utilize observations on laterality and handedness to gain insight into the initiating mechanism of monozygotic human quadruplets. STUDY DESIGN: In a study of thumbprints and handedness in sets of monozygotic quadruplets, homolateral and heterolateral comparisons were used to distinguish mirror-image from symmetric divisions. RESULTS: Data were consistent with a mechanism involving a mirror-image duplication in the first division and a subsequent symmetric separation of the daughters. In addition, these data failed to substantiate a genetic linkage between dermatoglyphics and handedness. CONCLUSION: Reported and reviewed data tend to support the analogy of monozygotic polyembryony in armadillos and humans. The plausible roles of delayed implantation and intercellular bonding are explored, especially as they relate to ambient calcium concentration. Factors tending to enhance the likelihood of monozygotic twinning include calcium channel blockers, lactation, ovulation induction and prolonged in vitro fertilization incubation. PMID- 11396376 TI - Natural history of twin-twin transfusion syndrome. AB - OBJECTIVE: To determine the natural history of pregnancies with twin-twin transfusion syndrome (TTTS). STUDY DESIGN: All cases of TTTS at our institution since 1991 and in a MED-LINE search since 1966 were retrospectively reviewed. The prenatal diagnosis of TTTS required the presence of monochorionic/diamniotic placentation (absence of twin-peak sign, thin membrane, single placenta, same sex) and of polyhydramnios (largest pocket > 8 cm) in one sac and oligohydramnios (largest pocket < 2 cm or stuck twin) in the other. Only cases diagnosed at < or = 28 weeks were included. Upon diagnosis, all patients were counseled as to the availability, risks and benefits of serial amniocentesis, laser therapy, septostomy, umbilical cord ligation, and other medical and surgical interventions. RESULTS: Of 29 pregnancies identified at our institution with the above strict criteria for TTTS, 5 (17%) declined in utero therapy and were managed expectantly. The mean gestational age at presentation was 19.2 weeks (range, 16-28) and at delivery, 33.8 weeks (range, 22-41). Four (40%) of the twins survived past the neonatal period, and of the three with follow-up, all are free of neurologic sequelae despite the death in utero of their cotwin. From the literature, 136 fetuses with TTTS as defined above and managed expectantly were identified; 37 (27%) survived; 75% (9/12) of survivors with follow-up are neurologically normal. The most recent four studies have reported survival of untreated cases of 50% (17/34) when diagnosed at < or = 26 weeks and of 63% (20/32) when diagnosed at < or = 28 weeks. CONCLUSION: The perinatal survival of TTTS pregnancies managed without in utero procedures is approximately 30% overall and 63% in the four most recent series when diagnosed at < or = 28 weeks. The success of in utero therapeutic intervention should be assessed by randomized studies or at least compared to that in similar cases managed without such intervention. PMID- 11396377 TI - Clinical pregnancy rates in an IVF program. Use of the flare-up protocol after failure with long regimens of GnRH-a. AB - OBJECTIVE: To examine the efficacy of flare-up protocols in patients who failed to respond to a long protocol using gonadotropin-releasing hormone agonist (GnRH a). STUDY DESIGN: In this retrospective study, a total of 144 patients who underwent an in vitro fertilization/embryo transfer cycle with the flare-up protocol were analyzed. Of these, 111 patients (group I) had not responded well to ovulation induction with long-term down-regulation with GnRH-a (long protocol), and 33 (group II) had responded to ovulation induction with at least four follicles but failed to conceive after embryo transfer. The average age of the patients were 37.3 +/- 3.9 years (range, 28-43) and 36.5 +/- 3.7 (range, 24 44), respectively. All patients underwent a flare-up protocol with GnRH-a (leuprolide acetate, 0.5 mg/d) on day 2 and at least 6 ampules of gonadotropins on day 3 within 6 months following failure with the long protocol. Unresponsiveness was defined as having fewer than three developing follicles on day 7, with an estradiol level < 200 pg/mL. Patients with a cycle day 2 follicle stimulating hormone (FSH) level > 15 mIU/mL before initiating GnRH-a were not included in the flare-up protocol. The average day 2 FSH levels of the patients on the flare-up protocol cycles were 9.1 +/- 3.0 in group I and 7.1 +/- 2.0 in group II. RESULTS: In group I, 44 of 111 (39.6%) patients did not respond to the flare-up protocol even with an increased dose of gonadotropins. In 67 patients, an average of 7.2 +/- 2.3 oocytes were obtained. Embryo transfer was performed on 64 patients with an average of 3.2 +/- 0.6 embryos. Fertilization did not occur in three patients. Eleven women conceived, eight of them miscarried, and three delivered. The pregnancy rate per cycle initiated was 9.9% (11/111) and live birth rate per cycle initiated, 2.5% (3/111). In group II, 3 of 33 patients were cancelled. The reasons were inadequate ovarian response, risk of hyperstimulation and absence of oocytes after aspiration. In 28 patients the average number of oocytes obtained was 6.9 +/- 3.9, and the average number of embryos replaced was 2.7 +/- 1.0. Six patients conceived, and all miscarried. The pregnancy rate per cycle initiated was 6/33 (18.1%), and no live births were achieved. CONCLUSION: Although the flare-up protocol after an unsuccessful luteal phase long protocol increases the pregnancy rate per cycle slightly, the live birth rate is not improved in poor responders. PMID- 11396378 TI - Incidental finding of malignant mixed mesodermal tumor at hysterectomy for uterine prolapse. A case report. AB - BACKGROUND: The finding of unanticipated pathology in a uterus after vaginal hysterectomy for prolapse is uncommon. CASE: An incidental small malignant mixed mesodermal tumor was found at vaginal hysterectomy in a 68-year-old woman. CONCLUSION: A MED-LINE search found no other reported cases of malignant mixed mesodermal tumor in a patient undergoing vaginal hysterectomy for uterine prolapse. Unexpected endometrial and cervical lesions will be discovered occasionally after hysterectomy for benign disease. PMID- 11396379 TI - Laparoscopic management of Sertoli-Leydig cell tumors of the ovary. A report of two cases. AB - BACKGROUND: Sertoli-Leydig cell tumor is a rare ovarian tumor with an incidence of < .5% of all ovarian tumors. Laparotomy is the standard approach to these cases. CASES: Sertoli-Leydig cell tumors were diagnosed in two young, nulliparous, infertile women. Both presented with secondary amenorrhea. Virilization was found in one. Their testosterone levels were high, and sonography revealed a solid, echogenic mass in the fornix. Laparoscopic removal was performed. Both women achieved normal menstruation one month after the operation, and one became pregnant and gave birth to a healthy infant. CONCLUSION: There are very few case reports of laparoscopic removal of such tumors. Laparoscopic surgery, which is minimally invasive and cosmetically acceptable and has a speedy recovery, should be the approach of choice for these patients. PMID- 11396380 TI - Trisomy 13 in one fetus from a twin gestation after intracytoplasmic sperm injection. A case report. AB - BACKGROUND: Intracytoplasmic sperm injection (ICSI) is an assisted reproductive technique used in the treatment of severe male factor infertility. Reports of chromosomal aberrations in some fetuses conceived following ICSI have raised concern that the technique may contribute to the development of genetic disease. CASE: Trisomy 13 occurred in one of twin fetuses of a 28-year-old woman who underwent ICSI. This case is unique because it is the first reported one of an autosomal trisomy complicating one fetus from a twin gestation conceived by ICSI. It is also the first case describing an autosomal trisomy following ICSI in a woman under the age of 35. CONCLUSION: Chromosomal abnormalities associated with ICSI may be accounted for by factors other than advanced maternal age. Chromosomal aberrations may be related to the procedure itself. Therefore, patients should be advised of the possibility of aneuploidy in association with ICSI, and genetic amniocentesis should be considered regardless of maternal age. PMID- 11396381 TI - Prolapsed cervical myoma after uterine artery embolization. A case report. AB - BACKGROUND: Cervical prolapse of myomas following uterine artery embolization is an infrequently reported occurrence. CASE: A 40-year-old woman with a history of three prior abdominal myomectomies underwent uterine artery embolization for severe menorrhagia due to myomas. Two weeks later a large cervical myoma prolapsed, requiring an abdominal hysterectomy. CONCLUSION: Cervical myomas should be considered a relative contraindication to uterine artery embolization. PMID- 11396383 TI - Urethral diverticulum in women. PMID- 11396382 TI - Craniopharyngioma and Bardet-Biedl syndrome. A case report. AB - BACKGROUND: Bardet-Biedl syndrome is a rare disorder and associated with a variety of anomalies. CASE: An 18-year-old woman was referred with primary amenorrhea. Following physical, ophthalmologic, psychiatric, hormonal and radiologic examinations, the diagnosis of both craniopharyngioma and Bardet-Biedl syndrome was established. CONCLUSION: Although the pathogenesis of hypogonadism in a woman with Bardet-Biedl syndrome remains unclear, cranial structures, especially the hypothalamus and pituitary gland, should be investigated to reveal any possible abnormalities. PMID- 11396384 TI - Managing anemia and blood loss in elective gynecologic surgery patients. AB - Hysterectomy is the second-most-common surgical procedure among premenopausal women. The conditions that lead to the need for a hysterectomy often are accompanied by chronic blood loss that can lead to anemia. Moreover, hysterectomy and myomectomy may result in significant blood loss, which exacerbates the anemia. The presence of fatigue associated with anemia has a substantially negative impact on quality of life and the ability to perform activities of daily living. Options for alleviating perioperative anemia include minimizing surgical blood loss, blood transfusion, supplementation with hematinics, such as iron and folic acid, and treatment with recombinant human erythropoietin. Treating preoperative anemia is expected to help correct anemia prior to surgery and may have a positive impact on anemia-related symptoms and surgical outcomes. PMID- 11396385 TI - Quality-of-life assessment in gynecologic surgery. AB - More than 90% of gynecologic surgery is performed for nonmalignant conditions, with a major objective of improving the patient's health-related quality of life (QOL). Clinical studies and patient surveys demonstrate that fatigue, diminished energy levels, increased need for rest, delayed time to return to work, difficulty performing daily routines, and difficulty caring for family and home persist for weeks to months or more following surgery. The social and economic implications of these outcomes provide a rationale for improving the QOL of gynecologic patients in the early weeks of recovery from surgery. Persistent and debilitating fatigue, which can lead to diminished QOL, is even more common than pain following hysterectomy. Global and specific subjective self-assessment instruments have been developed to measure fatigue as well as QOL parameters in postoperative gynecologic surgery patients. In addition, a QOL instrument combining both subjective self-assessment scales and objective measures of hemoglobin, hematocrit and muscle strength has been validated in postoperative orthopedic patients and may also have application in gynecologic surgery patients. Collectively, these various instruments may be useful in the assessment of recuperative power and vitality during early postoperative recovery in patients undergoing gynecologic surgery. PMID- 11396386 TI - Erythropoietin, the biology of erythropoiesis and epoetin alfa. An overview. AB - Erythropoietin, a glycoprotein hormone, is synthesized predominantly in the kidney and secreted by renal cortical interstitial cells in response to tissue hypoxia. Erythropoietin is the main regulator of the production of red blood cells. It functions in the recruitment and differentiation of erythroid progenitor cells and aids in their maintenance and survival. Erythropoietin also stimulates the synthesis of hemoglobin. In the last 15 years, the ready availability of recombinant human erythropoietin (r-HuEPO, epoetin alfa) has permitted the clinical investigation and application of this hormone to the treatment of anemia in various patient populations. Epoetin alfa has been shown to accelerate erythropoiesis and reduce allogeneic blood transfusion in major elective, noncardiac, nonvascular surgery and in certain anemic patients with chronic renal failure, nonmyeloid malignancies and human immunodeficiency virus infection. In addition to improving hematologic parameters, epoetin alfa therapy can enhance health-related quality of life in these patients. The success of epoetin alfa in treating anemia in other surgical populations suggests that it may be of benefit in treating the perioperative anemia that is highly prevalent in gynecologic surgery patients. Further investigation of the use of epoetin alfa in patients undergoing gynecologic surgery would increase awareness of its benefits for this patient population. PMID- 11396387 TI - Clinical experience with epoetin alfa in the management of hemoglobin levels in orthopedic surgery and cancer. Implications for use in gynecologic surgery. AB - Recombinant human erythropoietin (r-HuEPO, epoetin alfa) is used for treatment of anemia associated with chemotherapy for non-myeloid malignancies, chronic renal failure and zidovudine treatment in patients infected with the human immunodeficiency virus and for anemic patients undergoing elective, noncardiac, nonvascular surgery. Epoetin alfa has been shown to safely increase preoperative hemoglobin (Hb) levels in anemic patients undergoing elective noncardiac, nonvascular surgery and is more effective than preoperative autologous blood donation in reducing the need for perioperative blood transfusions in orthopedic surgery patients. Epoetin alfa was shown to significantly increase Hb levels and decrease transfusion requirements in gynecologic cancer patients undergoing chemotherapy. A once-weekly regimen of 40,000 IU per dose was effective in these patients. In addition to decreasing transfusion requirements and increasing Hb, epoetin alfa for relieving anemia-related fatigue and improving quality of life was demonstrated in clinical trials in anemic cancer patients receiving chemotherapy. With regard to quality of life in orthopedic surgery patients, a novel instrument to measure the effect of Hb management on postoperative recuperative power (i.e., vigor, functional ability) has been validated and may prove to be useful in optimizing rehabilitation and discharge planning. Extensive clinical experience with epoetin alfa in anemic patients undergoing major elective orthopedic surgery or those with gynecologic cancer provides a strong basis for its use in gynecologic surgery. PMID- 11396388 TI - Epoetin alfa use in gynecology. Past, present and future. AB - Perioperative anemia is a common complication of major surgery that may lead to prolonged and debilitating fatigue and reduction in health-related quality of life (QOL). Treatment with recombinant human erythropoietin (r-HuEPO, epoetin alfa) has been shown to increase perioperative hemoglobin (Hb) and hematocrit (HCT) levels, thereby facilitating postoperative recovery in orthopedic surgery patients. Treatment with epoetin alfa has also been shown to increase Hb and HCT levels and improve QOL in anemic cancer patients undergoing chemotherapy. The clinical and QOL benefit of using epoetin alfa in these patient populations provides the rationale for its use in patients undergoing gynecologic surgery. Because persistent fatigue is the most common complaint of patients following hysterectomy, the use of epoetin alfa should be considered to preoperatively correct anemia in this patient population. Research has been initiated to increase our understanding of the role of epoetin alfa in treating anemic patients (Hb levels < or = 13 g/dL) undergoing surgery for benign gynecologic disease, especially as it relates to postoperative QOL. Future studies should investigate the use of epoetin alfa in patients with gynecologic cancers. These studies should confirm the role of epoetin alfa in combination with iron supplementation to improve perioperative Hb/HCT levels and overall QOL in patients undergoing gynecologic surgery. PMID- 11396389 TI - Acute complications of sickle cell disease in children. AB - Sickle cell disease is a recessively inherited condition in which synthesis of haemoglobin is abnormal. The disease, which occurs mainly in people of African, African-Caribbean, Indian, Mediterranean and Middle Eastern descent, is characterised by chronic anaemia, susceptibility to infection, bouts of severe pain and organ dysfunction. While the life expectancy for patients has improved, from a median survival age of 14 years in the 1970s among those homozygous for the sickle haemoglobin gene to survival into the mid-40s, childhood remains a period of peak mortality and morbidity. Here, we discuss the acute complications of sickle cell disease in children, concentrating on the management of the acutely unwell child. PMID- 11396390 TI - Lowering blood pressure in particular patient groups. AB - In certain groups of patients, treatment of high blood pressure raises particular problems. These groups include, for instance, older people, those with co existing disease (such as renal disease or diabetes), those of certain racial origin and pregnant women. Here, we discuss the use of antihypertensive drugs in such groups. PMID- 11396391 TI - The influence of contour line size and location on the reproducibility of topographic measurement with the Heidelberg Retina Tomograph. AB - BACKGROUND: The recommended contour line (CL) location with the Heidelberg Retina Tomograph (HRT) is on the inner edge of Elschnig's scleral ring. This study investigated HRT parameter reproducibility when: (i) the CL size is altered relative to Elschnig's ring; (ii) the CL is either redrawn or imported between images. METHODS: Using the HRT, seven 10 degrees images were acquired for 10 normal volunteers and 10 primary open angle glaucoma (POAG) subjects. A CL was drawn on one image for each subject using Elschnig's scleral ring for reference and imported into subsequent images. The CL diameter was then (a) increased by 50 microns; (b) increased by 100 microns; and (c) decreased by 50 microns. To investigate the effect of the method of contour line transfer between images a CL was: (1) defined for one image and imported to 6 subsequent images; (2) drawn separately for each image. RESULTS: Parameter variability improved as the size of the CL increased for the normal group relative to Elschnig's ring but was unchanged in the POAG group. The export/import function (method 1) resulted in better parameter reproducibility than the redrawing method for both groups. CONCLUSIONS: The exporting and importing function resulted in better parameter variability for both subject groups and should be used for transferring CLs across images for the same subject. Increasing the overall CL size relative to Elschnig's scleral ring improved the reproducibility of the measured parameters in the normal group. No significant difference in parameter variability was observed for the POAG group. This suggests that the reproducibility of HRT images are affected more by the variation in topography between images than change in CL definition. PMID- 11396392 TI - The ChromaGen contact lens system: colour vision test results and subjective responses. AB - The ChromaGen lens system is designed to enhance colour perception in colour vision deficiency (CVD). To investigate its efficacy, 14 CVD subjects were prescribed ChromaGen contact lenses. Colour vision tests (Ishihara, Farnsworth Munsell D-15, Farnsworth Lantern) were administered at baseline, lens dispensing, and after a 2-week lens-wearing trial during which subjective responses were recorded daily using visual analogue scales. ChromaGen lenses significantly reduced ishihara error rates (p < 0.001; ANOVA), particularly for deutan subjects. There was also a significant reduction in errors (p < 0.005) on the D 15 test. Conversely, lens wear had no significant effect on Farnsworth Lantern test performance. Subjectively, subjects reported enhanced colour perception, but poor vision in dim light. Judgement of distance and motion were only slightly affected. We conclude that ChromaGen lenses may enhance subjective colour experience and assist in certain colour-related tasks, but are not indicated as an aid for CVD in occupations with colour vision-related restrictions. PMID- 11396393 TI - Influence of chronic alcohol abuse and ensuing forced abstinence on static subjective accommodation function in humans. AB - PURPOSE: Acute alcohol ingestion can change accommodation, but the long term effects of sustained alcohol consumption on accommodative function have not been studied in detail. This study was thus undertaken on individuals with a history of alcohol abuse. METHODS: Thirty-seven male individuals aged 25-56 years (average 40 years) from an alcohol rehabilitation centre in Inverness, Scotland, were assessed on admission and after a week of forced abstinence. The results were compared to a paired age-matched set of control male subjects. The static amplitude of accommodation was measured by an RAF rule, and the pupil size measured with a pupil gauge. RESULTS: On admission, the group mean measured amplitude of accommodation was 4.7 +/- 2.2 D (mean +/- SD). These values for the alcoholics were lower than age-matched controls (of 5.9 +/- 2.9 D). The slope of the age-dependent decline in RAF rule accommodation measures was significantly smaller for the alcoholics compared to controls (at 0.215 +/- 0.027 D/year versus 0.332 +/- 0.015 D/year, respectively; p < 0.001), with the younger alcoholics showing a greater impairment. Following abstinence, there was no measurable change in accommodation measured, indicating the lower amplitude in the alcoholics was not attributable to circulatory alcohol levels. The resting pupil diameter in the alcoholics was 4.37 +/- 0.63 mm compared to the controls of 3.97 +/- 0.75 mm, with a higher incidence of small pupils (< or = 3 mm) in the controls. CONCLUSIONS: The results indicate that chronic alcohol use can adversely affect subjective static accommodation, especially in younger alcoholics, as well as cause slight mydriasis. PMID- 11396395 TI - The effect of monochromatic aberrations on Autoref R-1 readings. AB - The Canon Autoref R-1 is used extensively to investigate aspects of the accommodation response of the human eye. The measurement zone of the Autoref R-1 is known to lie outside the central pupil and the instrument's readings are therefore potentially influenced by the monochromatic aberrations of the eye. Since these aberrations are known to change as a function of accommodation, the error associated with the aberrations might influence the stimulus-response slope. We used masks and contact lenses with known amounts of spherical aberration to establish the region of the pupil used by the Autoref R-1 in deriving a reading of the refractive power of the eye. This region was estimated to be narrowly centred on a 3.5-mm diameter. The spherical aberration of the eye was measured for five subjects across a range of accommodation levels and used to predict the likely effect upon Autoref R-1 readings. Positive spherical aberration will result in an apparent lead of accommodation, while negative spherical aberration causes an apparent lag of accommodation. For eyes with normal levels of spherical aberration, the slope of the stimulus-response will appear to be marginally flatter when measured with the Autoref R-1. An example of an eye with coma-like aberrations is also shown to produce Autoref R-1 results that will differ significantly from more central refractive measurements. PMID- 11396394 TI - The effect of perceived length on visuomotor localization. AB - PURPOSE: The purpose of this experiment was to study visuomotor localization in the presence of either a horizontal array of equally spaced dots or a thin horizontal line. METHODS: Pointing behavior was used to assess directional localization. In experiment 1, subjects were made myopic using a contact lens and then corrected with a spectacle lens. Subjects were tested in the presence and absence of a regularly spaced, horizontal array of dots with and without the contact lens/spectacle combination. In experiment 2, subjects wore the contact lens/spectacle in all cases. Some subjects were tested in the presence and then in the absence of a regularly spaced, horizontal array of dots while the order of conditions was reversed for other subjects. In experiments 3 and 4, subjects were tested without the contact lens/spectacle combination. In experiment 3, subjects were tested in the absence and then in the presence of a regularly spaced, horizontal arrays of dots. In experiment 4, subjects were tested in the absence and then in the presence of a thin horizontal line. RESULTS: In experiment 1, in the absence of the array of dots, subjects undershot targets with the contact lens/spectacle combination. When the array was present, pointing with the contact lens/spectacle combination was accurate. In experiment 2, subjects undershot targets in the absence of the array of dots if this condition was performed first. If the array was present in the initial condition, the pointing undershoot in the second condition (array absent) was reduced. In all cases, the pointing undershoot was reduced in the presence of the array. In experiments 3 and 4, a pointing overshoot was found in the presence of an array of dots or a thin line. CONCLUSIONS: It is concluded that extraretinal eye position information is not the primary determinant of visuomotor localization in the presence of a horizontal contour. The overshoot produced by a horizontal contour may be related to a length illusion brought about by spatial filtering in the visual system or inaccurate distance judgments. PMID- 11396396 TI - Intraocular pressure after excimer laser myopic refractive surgery. AB - The aim of this study was to determine whether intraocular pressure (IOP), as measured by Goldmann applanation or non-contact tonometry, shows systematic changes in patients who have undergone photorefractive keratectomy (PRK) or laser in situ keratomileusis (LASIK). IOP was measured by central Goldmann and non contact tonometry in 54 patients pre and post-PRK, and in 43 patients pre- and post-LASIK. An interval of 12 months was allowed after surgery. Patients were selected to have one of four specific initial values of refractive error (-2.5, 5.0, -7.5 and -10.0 D). Fellow unoperated eyes were used as controls. A paired Student's t-test and a one-way ANOVA test were used for statistical analysis. After PRK and LASIK, a statistically significant decrease (p < 0.01) was observed in the IOP of the treated eyes (but not for control eyes; p > 0.01). Although the magnitude of the change increased with the attempted refractive correction, this trend was not statistically significant (p > 0.01). No statistically significant differences were found between the results obtained following the two types of surgery, although the recorded fall in IOP was smaller following LASIK (p > 0.01). The IOP measured after PRK and LASIK for myopia may be reduced because of reduced corneal thickness and curvature and, possibly, tissue softening after natural healing. The presence or absence of Bowman's membrane does not appear to be important in this context. The reduction in measured IOP following refractive surgery, by about 0.5 mmHg/D of myopic correction, needs to be remembered when possible abnormality of IOP in such patients is being considered. PMID- 11396397 TI - Rigid gas permeable contact lens and corneal topography. AB - We investigated the effect of high Dk daily wear rigid gas permeable contact lenses on corneal topography. Eight young myopic subjects wore hard contact lenses for 21 days. Corneal topography was monitored using the EyeSys system. It was measured every day during the next 21 days after the fitting. We recorded the corneal radius of curvature at 16 peripheral points approximately 1, 2, 3 and 4 mm along the four principal meridians (nasal, superior, inferior and temporal). Our study showed that daily wear RGP Forum 210 does not produce significant alterations of the corneal curvature as a function of time. PMID- 11396398 TI - The pattern electroretinogram (PERG) with contralateral corneal reference. AB - The pattern electroretinogram (PERG) is a sensitive test for investigation of the proximal retina. For monocular recording, the contralateral corneal reference (CCR) electrode position has been suggested as a possible alternative to the conventional ipsilateral temporal reference (ITR). We therefore compared the effect of these electrode positions on 26 subjects (median age 45.5 years, IQR 24.5-61.5). The signals were recorded monocularly with DTL nylon fibre electrodes. The visual stimuli were high contrast 19' black-white checkerboard patterns, reversing 4 times/s. PERG responses obtained from the CCR gave larger amplitudes and the increase in amplitude was statistically significant (P < 0.001). Because the CCR signal is larger, new normative ranges must be established to avoid false negative results. PMID- 11396399 TI - Interconverting the matrix and principal meridional representations of dioptric power in general including powers with nonorthogonal and complex principal meridians. AB - The principal meridians of the powers of thick astigmatic systems, like the eye, are not necessarily at right angles. The consequence is a class of phenomena included in the category commonly described as irregular astigmatism. The conventional principal meridional representation of power, however, is unsuited to quantitative analysis. This paper presents equations for converting from the principal meridional form of power to a representation, the dioptric power matrix, which is amenable to quantitative analysis. It generalizes an earlier paper which treated only powers of a conventional form in which the principal meridians are always at right angles. It copes in particular with what are known as asymmetric powers. A routine is also presented for converting in the reverse direction, from the power matrix to the principal meridional form of power. The principal meridional form of power turns out not always to be unique, there being distinct powers (they are asymmetric) with the same principal powers and meridians. Thus, in general, the dioptric power matrix is a satisfactory representation of power while the principal meridional representation is not. PMID- 11396400 TI - Interpretating nonorthogonal keratometric measurements. PMID- 11396401 TI - [Clinical study of zirconium oxide bridges in the posterior segments fabricated with the DCM system]. AB - Today's dental reconstructive therapeutic concepts require restoration of high esthetic quality and excellent biocompatibility. Full ceramic reconstructions accomplish these requirements but only for anterior teeth and premolars. For all ceramic bridges the mechanical strength was insufficient to withstand the posterior chewing forces. Frequently the interdental connectors cracked, and the only way to prevent these fractures was to overconture the connectors to a size of approx. 16 mm2. The high-tech ceramic zirconia is a potential alternative for three-to five-unit full ceramic bridges in the functionally loaded posterior segments. Experimental zirconia bridges which were fabricated using the DCM system (Direct Ceramic Machining System at the ETH Zurich, were tested in vitro. The frameworks were digitally enlarged by 20% and were easily milled from a presintered yet porous zirconia blank. After the milling process, the framework was densely sintered and shrank to its original size. Due to these positive in vitro results a clinical investigation was started. 22 veneered zirconia bridges were luted; 19 molars and 25 premolars were prepared. The connectors, max. 7 mm2, of all these bridges, have been functionally loaded by antagonists. After a mean observation time of 385 days (307 days to 488 days), all 22 bridges did not show any cracks in the framework or in the veneering porcelain. The patients commented particularly on the low heat conduction rate of zirconia. The only endodontic problem which occurred could not be directly connected to the type of bridge framework. The reliability of zirconia bridges in this investigation was connected to the DCM-Process. No statement about other zirconia-systems can be made on the results of this study. PMID- 11396403 TI - Validation of publication of new names and new combinations previously effectively published outside the IJSEM. PMID- 11396402 TI - [Resilience behavior of titanium implants with integrated maintenance-free biokinetic elements]. AB - Good functional properties are essential in dental implantology. Bio-kinetics elements are imitating dental resilience. In this study a new kind of implants with maintenance-free shock-absorbing elements was introduced and their bio mechanic properties were tested. The mobile implant (SIS Inc, Klagenfurt, Austria) is a self-cutting conical screw implant with an integrated bio-kinetic element. The shock absorber is a central part of the implant and a titanium ring obturates the shock absorbing unit in the implant. The resilience of the implant was tested by axial and excentric loading in a special testing unit. Furthermore a survival test of the elastic titanium ring in the most exposed cervical part of the implant was performed. The region was examined by scanning electron microscopy after 12 million loading cycles in the axial and radial direction. A progressive shock absorption was registered during radial and axial loading. The maximum movements were 0.06 mm in the axial and 0.16 mm in the radial direction. No signs of material destruction were seen in the electron microscopic analysis. Thus a maintenance-free bio-kinetic implant with progressive shock-absorbing qualities is presented. PMID- 11396404 TI - Notification that new names and new combinations have appeared in volume 50, part 5, of the IJSEM. PMID- 11396405 TI - Proceedings of the 11th European Bioenergetics Conference. Sussex, United Kingdom, 9-14 September 2000. PMID- 11396406 TI - Web alert. PMID- 11396407 TI - Web alert. PMID- 11396408 TI - Web alert. PMID- 11396409 TI - Web alert. PMID- 11396410 TI - Web alert. PMID- 11396411 TI - Teaching in China. PMID- 11396412 TI - Communication with the dentist is essential for small laboratories. PMID- 11396413 TI - Telemarketing. A cost effective way to gain accounts. PMID- 11396414 TI - Why can't I find the people I need? PMID- 11396415 TI - Combination chemotherapy for extensive small cell lung cancer -- experience from India. PMID- 11396416 TI - Yes! Time for global strategy to improve the treatment of lung cancer is right now. PMID- 11396417 TI - Construction of DNA restriction maps based on a simplified experiment. AB - MOTIVATION: A formulation of a new problem of the restriction map construction based on a simplified digestion experiment and a development of an algorithm for solving both ideal and noisy data cases of the introduced problem. RESULTS: A simplified partial digest problem and a branch and cut algorithm for finding the solution of the problem. PMID- 11396418 TI - [Prolonged fever and pain in the right upper quadrant in an elderly woman]. PMID- 11396419 TI - [Meningoencephalitis with cerebellar involvement associated with Coxiella burnetii pneumonia]. PMID- 11396421 TI - Cajal and consciousness: scientific approaches to consciousness on the centennial of Ramon y Cajal's Textura. Proceedings of a meeting. November 29-December 1, 1999. Zaragoza, Spain. PMID- 11396420 TI - Endolymphatic hydrops induced by chronic administration of vasopressin. PMID- 11396422 TI - Dose and dose rate effect in endovascular brachytherapy. PMID- 11396423 TI - Histological demonstration of coronary dissection leading to double luminal artery in swine model. PMID- 11396425 TI - Another useful index of ventricular function in patients with single ventricle. PMID- 11396424 TI - Atrial fibrillation and abnormalities of hemostatic factors. PMID- 11396426 TI - Niacin in patients with diabetes mellitus and coronary artery disease. PMID- 11396427 TI - Diabetes: risk factor for the development of pancreatic cancer or manifestation of the disease? AB - The objective of this study was to summarize the English-language literature that addresses diabetes mellitus as a potential risk factor for the development of pancreatic cancer. Studies exploring the relation between diabetes and pancreatic cancer published between 1970 and 1999 were identified from a MEDLINE search and are summarized. They showed that diabetes mellitus occurs more frequently in patients with pancreatic cancer than in the general population. Most of the literature supports the view that diabetes of at least 5 years' duration increases the risk of subsequently developing pancreatic cancer. In addition, experimental forms of diabetes clearly affect the induction and growth of pancreatic cancer. The currently available literature thus suggests that diabetes is a risk factor for the subsequent development of pancreatic cancer. PMID- 11396428 TI - Paradoxical embolism: diagnosis and management. PMID- 11396429 TI - Lack of efficacy of benzodiazepines in treating gamma-hydroxybutyrate withdrawal. PMID- 11396430 TI - Acetaminophen toxicity nomogram error: Barkin's Textbook of Pediatric Emergency Medicine. PMID- 11396431 TI - Summaries for patients. Long-term outcome of patients with kidney damage due to Goodpasture disease wo receive very intensive treatment. PMID- 11396432 TI - Summaries for patients. Unrecognized heart attacks in women with known coronary artery disease. PMID- 11396433 TI - Summaries for patients. Treatment for sleep apnea in people without symptoms. PMID- 11396434 TI - Summaries for patients. Home monitoring service improves blood pressure control in patients with high blood pressure. PMID- 11396435 TI - An appreciation of Dr. Gordon Jackson Rees FRCA, FRCP, FRCPCH: pioneer of paediatric anaesthesia. PMID- 11396436 TI - Correlation between plasma endotoxin, plasma cytokines, and plasminogen activator inhibitor-1 activities in septic patients. AB - The objective of this study was to evaluate the relation between the clinical and plasma parameters and the changes in plasma endotoxin activity with 2 hours of endotoxin-adsorbing therapy using polymyxin B (PMX). A total of 88 consecutive patients were admitted for PMX treatment of severe sepsis or septic organ failure. Standard supportive care was continued without alteration during PMX treatment. Endotoxin, tumor necrosis factor-alpha (TNFalpha), interleukin-6 (IL 6), IL-10, and plasminogen activator inhibitor-1 (PAI-1) activities and clinical parameters were measured before, immediately after, and the day after PMX treatment. The mean APACHE II and III scores were 24.2 +/- 1.0 and 85.8 +/- 3.0, respectively. The 2-week survival rate was 51.1%. In survivors, TNFalpha, IL-6, IL-10, and PAI-1 activities were significantly decreased during the 2-hour PMX treatment, the following day, or both times. There was no significant change in the parameters, except for TNFalpha, after PMX in nonsurvivors. In the subgroup whose plasma endotoxin decreased more than 30%, IL-6, TNFalpha, and PAI-1 significantly decreased after 2 hours of PMX or the following day (or both), but all four parameters in nonsurvivors showed no significant change. Hence PMX adsorbed plasma endotoxins and contributed to reductions in plasma proinflammatory cytokine levels and to improved clinical parameters during the 2 hour treatment. Changes in these parameters correlated with changes in plasma endotoxin activity in survivors whose plasma endotoxin levels were adequately reduced. PMID- 11396437 TI - Clinical impact of continuous renal replacement therapy on multiple organ failure. AB - Because continuous renal replacement therapy (CRRT) may enhance inflammatory mediator removal, this review assesses its impact on multiple organ failure (MOF). Regarding MOF with acute renal failure (ARF), the overall mortality of 2313 CRRT patients (43 studies) was 62.8% compared with 59.1% (p = 0.046) in 961 intermittent hemodialysis (IHD) patients (12 other studies). Of 13 CRRT studies with an IHD comparison group, 3 showed that the groups had a similar risk, but IHD mortality was higher; 1 noted that CRRT had lower mortality (risk not stated); and 4 showed similar mortality and greater CRRT risk. Aggregate mortality was IHD 69.5% and CRRT 63.9% (p = 0.02). Of the six studies with matched groups (age and APACHE II scores), IHD mortality was higher (70.9% vs. 60.1%, p = 0.01). CRRT pulmonary gas exchange, hemodynamic instability, azotemia control, fluid overload, and nutritional support were better. Regarding MOF without ARF, of 14 CRRT studies (14.5 patients per study), only 4 had comparison groups. Patient conditions were as follows: acute respiratory distress syndrome, six studies; sepsis, three studies; septic shock, two studies; pancreatitis, one study; critically ill patients, one study; and cardiac surgery with respiratory failure, one study. Of the three studies with a control group, the mortality was the same. There was minimal evidence that CRRT improved pulmonary gas exchange or hemodynamic instability. For MOF patients with ARF, there is compelling evidence that CRRT provides better survival than IHD and more improvement in pulmonary gas exchange, hemodynamic instability, azotemia control, fluid overload, and nutritional support. In patients with MOF and no renal failure, there is little evidence that CRRT enhances survival, oxygenation, or perfusion. Controlled trials demonstrating a CRRT benefit are necessary before CRRT can be recommended for MOF without ARF. PMID- 11396438 TI - Blood purification in the intensive care unit: evolving concepts. AB - Until relatively recently surgeons were familiar with the concept that some of their patients admitted to the intensive care unit require dialysis to deal with the development of severe acute renal failure. Under such circumstances the nephrology team would then attend the patient and take over that aspect of management. More recently, however, this situation has undergone a significant evolution because of the advent of continuous renal replacement therapy (CRRT). First introduced as "last ditch" therapy in the most critically ill patients who were hemodynamically intolerant of hemodialysis, CRRT has become more and more widely used. It is now the dominant form of artificial renal support in Australia and close to being the dominant treatment of the severe acute renal failure of critical illness in most European countries. The use of CRRT in the United States is rapidly growing. The arrival of CRRT has also renewed interest in the wider concept of blood purification during critical illness. Experimental and preliminary human data suggest that such blood purification therapies may indeed have beneficial immunomodulatory effects. Accordingly, CRRT is now being considered as a potential adjuvant treatment of septic shock and has even moved into the operating room as a tool for antiinflammatory therapy and volume control. The intensivist-surgeon and the general surgeon need to be aware of and understand these developments in extracorporeal therapy if they wish to make the full armamentarium of modern treatment available to their sickest patients. PMID- 11396439 TI - Nonviral transfer of the gene encoding coagulation factor VIII in patients with severe hemophilia A. AB - BACKGROUND: We tested the safety of a nonviral somatic-cell gene-therapy system in patients with severe hemophilia A. METHODS: An open-label, phase 1 trial was conducted in six patients with severe hemophilia A. Dermal fibroblasts obtained from each patient by skin biopsy were grown in culture and transfected with a plasmid containing sequences of the gene that encodes factor VIII. Cells that produced factor VIII were selected, cloned, and propagated in vitro. The cloned cells were then harvested and administered to the patients by laparoscopic injection into the omentum. The patients were followed for 12 months after the implantation of the genetically altered cells. An interim analysis was performed. RESULTS: There were no serious adverse events related to the use of factor VIII producing fibroblasts or the implantation procedure. No long-term complications developed, and no inhibitors of factor VIII were detected. In four of the six patients, plasma levels of factor VIII activity rose above the levels observed before the procedure. The increase in factor VIII activity coincided with a decrease in bleeding, a reduction in the use of exogenous factor VIII, or both. In the patient with the highest level of factor VIII activity, the clinical changes lasted approximately 10 months. CONCLUSIONS: Implantation of genetically altered fibroblasts that produce factor VIII is safe and well tolerated. This form of gene therapy is feasible in patients with severe hemophilia A. PMID- 11396440 TI - Increased need for thyroxine in women with hypothyroidism during estrogen therapy. AB - BACKGROUND: Women with hypothyroidism that is being treated with thyroxine often need higher doses when they are pregnant. Whether this need can be attributed solely to estrogen-induced increases in serum thyroxine-binding globulin or whether other factors are involved is not known. METHODS: In 11 postmenopausal women with normal thyroid function and 25 postmenopausal women with hypothyroidism treated with thyroxine, I assessed thyroid function before they started estrogen therapy and every 6 weeks for 48 weeks thereafter. The women with hypothyroidism included 18 women receiving thyroxine-replacement therapy and 7 women receiving thyrotropin-suppressive thyroxine therapy. On each occasion, serum thyroxine, free thyroxine, thyrotropin, and thyroxine-binding globulin were measured. RESULTS: In the women with normal thyroid function, the serum free thyroxine and thyrotropin concentrations did not change, whereas at 12 weeks the mean (+/-SD) serum thyroxine concentration had increased from 8.0+/-0.9 microg per deciliter (103+/-12 nmol per liter) to 10.4+/-1.5 microg per deciliter (134+/ 19 nmol per liter, P<0.001) and the serum thyroxine-binding globulin concentration had increased from 20.3+/-3.5 mg per liter to 31.3+/-3.2 mg per liter, P<0.001). The women with hypothyroidism had similar increases in serum thyroxine and thyroxine-binding globulin concentrations during estrogen therapy, but their serum free thyroxine concentration decreased from 1.7+/-0.4 ng per deciliter (22+/-5 pmol per liter) to 1.4+/-0.3 ng per deciliter (18+/-4 pmol per liter, P<0.001) and their serum thyrotropin concentration increased from 0.9+/ 1.1 to 3.2+/-3.1 microU per milliliter (P<0.001). The serum thyrotropin concentrations increased to more than 7 microU per milliliter in 7 of the 18 women in the thyroxine-replacement group and to more than 1 microU per milliliter in 3 of the 7 women in the thyrotropin-suppression group. CONCLUSIONS: In women with hypothyroidism treated with thyroxine, estrogen therapy may increase the need for thyroxine. PMID- 11396441 TI - Evidence that human cardiac myocytes divide after myocardial infarction. AB - BACKGROUND: The scarring of the heart that results from myocardial infarction has been interpreted as evidence that the heart is composed of myocytes that are unable to divide. However, recent observations have provided evidence of proliferation of myocytes in the adult heart. Therefore, we studied the extent of mitosis among myocytes after myocardial infarction in humans. METHODS: Samples from the border of the infarct and from areas of the myocardium distant from the infarct were obtained from 13 patients who had died 4 to 12 days after infarction. Ten normal hearts were used as controls. Myocytes that had entered the cell cycle in preparation for cell division were measured by labeling of the nuclear antigen Ki-67, which is associated with cell division. The fraction of myocyte nuclei that were undergoing mitosis was determined, and the mitotic index (the ratio of the number of nuclei undergoing mitosis to the number not undergoing mitosis) was calculated. The presence of mitotic spindles, contractile rings, karyokinesis, and cytokinesis was also recorded. RESULTS: In the infarcted hearts, Ki-67 expression was detected in 4 percent of myocyte nuclei in the regions adjacent to the infarcts and in 1 percent of those in regions distant from the infarcts. The reentry of myocytes into the cell cycle resulted in mitotic indexes of 0.08 percent and 0.03 percent, respectively, in the zones adjacent to and distant from the infarcts. Events characteristic of cell division -the formation of the mitotic spindles, the formation of contractile rings, karyokinesis, and cytokinesis--were identified; these features demonstrated that there was myocyte proliferation after myocardial infarction. CONCLUSIONS: Our results challenge the dogma that the adult heart is a postmitotic organ and indicate that the regeneration of myocytes may be a critical component of the increase in muscle mass of the myocardium. PMID- 11396442 TI - Treatment of the immune dysregulation, polyendocrinopathy, enteropathy, X-linked syndrome (IPEX) by allogeneic bone marrow transplantation. PMID- 11396443 TI - Images in clinical medicine. Disseminated penicillium marneffei infection in a patient with AIDS. PMID- 11396444 TI - AIDS--the first 20 years. PMID- 11396445 TI - The hemophilias--from royal genes to gene therapy. PMID- 11396446 TI - AIDS--will the next 20 years be different? PMID- 11396447 TI - Gene therapy for hemophilia. PMID- 11396448 TI - Estrogen, thyroxine binding in serum, and thyroxine therapy. PMID- 11396449 TI - High hopes for the heart. PMID- 11396450 TI - AIDS--past and future. PMID- 11396451 TI - Vaccinations and multiple sclerosis. PMID- 11396452 TI - Vaccinations and multiple sclerosis. PMID- 11396453 TI - Vaccinations and multiple sclerosis. PMID- 11396454 TI - Vaccinations and multiple sclerosis. PMID- 11396455 TI - Vaccinations and multiple sclerosis. PMID- 11396456 TI - Vaccinations and multiple sclerosis. PMID- 11396457 TI - Intracoronary radiotherapy for restenosis. PMID- 11396458 TI - Controlling tobacco use. PMID- 11396459 TI - Controlling tobacco use. PMID- 11396460 TI - Controlling tobacco use. PMID- 11396462 TI - Quality measures for the emergency services. Proceedings of a symposium. London, United Kingdom, 3 February 2000. PMID- 11396461 TI - HIV-associated coronary arteritis in a patient with fatal myocardial infarction. PMID- 11396466 TI - From the Centers for Disease Control and Prevention. Fatal and severe hepatitis associated with rifampin and pyrazinamide for the treatment of latent tuberculosis infection--New York and Georgia, 2000. PMID- 11396467 TI - From the Centers for Disease Control and Prevention. Severe malnutrition among young children--Georgia, January 1997-June 1999. PMID- 11396468 TI - From the Centers for Disease Control and Prevention. Prevalence of arthritis- United States, 1997. PMID- 11396469 TI - From the Centers for Disease Control and Prevention. Trends in blood lead levels among children--Boston, Massachusetts, 1994-1999. PMID- 11396470 TI - JAMA patient page. Health information on the Internet. PMID- 11396471 TI - Platelet-activating factor content in boar spermatozoa correlates with fertility. AB - The objective of this study was to evaluate the level of platelet-activating factor [1-O-alkyl-2-acetyl-sn-glycero-3-phosphorylcholine; PAF] content in spermatozoa between two groups of boars that differ in farrow rate percentages. The boar farrow rate was defined as High if it was > or = 70% and Low if it was < 70%. Fresh, extended semen was collected from sexually mature boars and used in the PAF extractions. Platelet-activating factor was detected in all semen samples assayed. The amount of PAF detected in spermatozoa obtained from the High group ranged from 1.90 to 11.30 pM/10(6) cells. The level of PAF in the Low group ranged from 0.92 to 4.96 pM/10(6) cells. Regression analysis revealed a positive (R2 = 0.369) and significant (P = 0.021) relationship between PAF content in boar spermatozoa and farrow rate. Spermatozoa-derived PAF levels (mean +/- SEM) were significantly higher (P = 0.015) in the High-farrow group (6.75 +/- 1.25 pM/10(6) cells) than in the Low-farrow group (2.45 +/- 0.51 pM/10(6) cells). The PAF content in spermatozoa was significantly higher (P = 0.035) in the High-average (> or = 10.5/litter) number of piglets born group (5.78 +/- 1.24 pM/10(6) cells) than in the Low-average (< 10.5/litter) number of piglets born group (3.34 +/- 1.19 pM/10(6) cells). Additionally, PAF content in spermatozoa was significantly higher (P = 0.034) in the High-average (> or = 9/litter) number of piglets born alive group (6.82 +/- 1.35 pM/10(6) cells) than the Low-average (< 9/litter) number of piglets born alive group (3.00 +/- 0.87 pM/10(6) cells). The data demonstrate that PAF is present in boar spermatozoa and that levels are significantly higher in individuals with a high-farrow rate status and high number of piglets born and born-alive. PMID- 11396472 TI - [Two hundred fifty veterinarians go to The Hague. Veterinarians protest against non-vaccine policy]. PMID- 11396473 TI - Direct and indirect antioxidant effects of nitric oxide: radically unsettled issues. PMID- 11396474 TI - EPR studies of nitric oxide interactions of alkoxyl and peroxyl radicals in in vitro and ex vivo model systems. AB - A model compound of lipid peroxidation, tert-butyl hydroperoxide (tBOOH), was used in vitro to investigate (i) the generation of tBOOH-derived free radicals by hematin or rat enterocytes and (ii) the modulation of cell-generated free radical production by a nitric oxide (NO) donor, or when these cells were primed to produce NO. In hematin-catalyzed decomposition of tBOOH, NO from nitrosoglutathione, or S-nitroso-N-acetylpenicillamine suppressed the generation of peroxyl radicals (measured by direct electron paramagnetic resonance) and tert butylalkoxyl, methoxyl, and methyl radicals (measured by electron paramagnetic resonance spin trapping). Similarly, co-incubation of S-nitroso-N acetylpenicillamine or nitrosoglutathione with tBOOH caused significant decreases in tBOOH-derived free radical generation catalyzed by enterocytes. Epithelial cells are the known source of the inducible form of NO synthase in the intestine of rats challenged with lipopolysaccharide (LPS). Enterocytes isolated from LPS treated rats produced decreased levels of tBOOH-derived radicals. These decreases in free radical production were further decreased when these cells were treated with LPS in vitro. These findings demonstrated that exogenously added or endogenously produced NO could modulate the extent of tBOOH-derived free radical generation in enterocytes. These decreases in free radical production could, at least in part, describe the protective role of NO from hydroperoxide-induced injury. PMID- 11396475 TI - Antioxidant mechanisms of nitric oxide against iron-catalyzed oxidative stress in cells. AB - Three distinct antioxidant pathways are considered through which iron-catalyzed oxidative stress may be regulated by nitric oxide (NO). The first two pathways involve direct redox interactions of NO with iron catalytic sites and represent a fast response that may be considered an emergency mechanism to protect cells from the consequences of acute and intensive oxidative stress. These are (i) NO induced nitrosylation at heme and non-heme iron catalytic sites that is capable of directly reducing oxoferryl-associated radicals, (ii) formation of nitrosyl complexes with intracellular "loosely" bound redox-active iron, and (iii) an indirect regulatory pathway that may function as an adaptive mechanism that becomes operational upon long-term exposure of cells to NO. In the latter pathway, NO down-regulates expression of iron-containing proteins to prevent their catalytic prooxidant reactions. PMID- 11396476 TI - Mechanisms of the antioxidant effects of nitric oxide. AB - The Janus face of nitric oxide (NO) has prompted a debate as to whether NO plays a deleterious or protective role in tissue injury. There are a number of reactive nitrogen oxide species, such as N2O3 and ONOO-, that can alter critical cellular components under high local concentrations of NO. However, NO can also abate the oxidation chemistry mediated by reactive oxygen species such as H2O2 and O2- that occurs at physiological levels of NO. In addition to the antioxidant chemistry, NO protects against cell death mediated by H2O2, alkylhydroperoxides, and xanthine oxidase. The attenuation of metal/peroxide oxidative chemistry, as well as lipid peroxidation, appears to be the major chemical mechanisms by which NO may limit oxidative injury to mammalian cells. In addition to these chemical and biochemical properties, NO can modulate cellular and physiological processes to limit oxidative injury, limiting processes such as leukocyte adhesion. This review will address these aspects of the chemical biology of this multifaceted free radical and explore the beneficial effect of NO against oxidative stress. PMID- 11396477 TI - Mechanisms of cell signaling by nitric oxide and peroxynitrite: from mitochondria to MAP kinases. AB - Many of the biological and pathological effects of nitric oxide (NO) are mediated through cell signaling pathways that are initiated by NO reacting with metalloproteins. More recently, it has been recognized that the reaction of NO with free radicals such as superoxide and the lipid peroxyl radical also has the potential to modulate redox signaling. Although it is clear that NO can exert both cytotoxic and cytoprotective actions, the focus of this overview are those reactions that could lead to protection of the cell against oxidative stress in the vasculature. This will include the induction of antioxidant defenses such as glutathione, activation of mitogen-activated protein kinases in response to blood flow, and modulation of mitochondrial function and its impact on apoptosis. Models are presented that show the increased synthesis of glutathione in response to shear stress and inhibition of cytochrome c release from mitochondria. It appears that in the vasculature NO-dependent signaling pathways are of three types: (i) those involving NO itself, leading to modulation of mitochondrial respiration and soluble guanylate cyclase; (ii) those that involve S-nitrosation, including inhibition of caspases; and (iii) autocrine signaling that involves the intracellular formation of peroxynitrite and the activation of the mitogen activated protein kinases. Taken together, NO plays a major role in the modulation of redox cell signaling through a number of distinct pathways in a cellular setting. PMID- 11396478 TI - Regulation of cyclooxygenase-2 expression by nitric oxide in cells. AB - Two different cyclooxygenases (COXs) are functional in mammals: COX-1 and COX-2. COX-2 is mainly an inducible isoform that shares significant features with inducible nitric oxide synthase (iNOS) in terms of its tissue distribution and participation in pathophysiological phenomena. Furthermore, the product of iNOS catalysis, nitric oxide (NO), is an important regulator of COX-2 activity and expression, and the products of COX-1 and COX-2 (diverse prostanoids) may also influence iNOS expression. Both positive and negative effects of NO on COX-2 expression have been encountered in experimental systems, showing that the outcome of the NO-COX-2 interaction is exquisitely dependent upon the temporal frame and the cell type studied. The pathophysiological significance of NO-COX cross-talk also arises from in vivo studies, in which most evidence points to a positive effect of NO on COX-2 activity and/or expression. This emphasizes the need to understand the underlying mechanisms. Among these, the capacity of NO and its effector cyclic GMP to modulate the function of several target proteins, including transcription factors such as nuclear factor-kappaB and activator protein-1, appears as the key pathway by which NO may regulate COX-2 expression. Given the capacity of some prostanoids to modulate the inflammatory response, the interplay between NO synthase and COX pathways stands at the center of the pathophysiological basis of inflammatory diseases. PMID- 11396479 TI - *NO, RSNO, ONOO-, NO+, *NOO, NOx--dynamic regulation of oxidant scavenging, nitric oxide stores, and cyclic GMP-independent cell signaling. AB - Following its release from nitric oxide synthase, nitric oxide seldom perfuses the cytosol; rather this reactive mediator quickly interacts with available target molecules proximate to its site of release. Within the cell, virtually every component, low-molecular-weight oxidants and reductants, proteins, lipids, sugars, and nucleic acids can be modified by nitrogen oxides thus acting as potential targets for reactive nitrogen oxides. Adducts formed by nitrogen oxides often modulate the cellular activities of the target molecules, and these modified molecules may be differentially metabolized or localized. The formation of nitrogen oxide adducts can be a reversible process, and the reactive nitrogen species released may be specifically oxidized or reduced during the process. Recently, numerous studies have demonstrated that reversible nitration of cellular proteins acts to transduce molecular signals regulating such diverse processes as muscle contraction, neurotransmission, protein metabolism, and apoptosis. The vast numbers of molecules that undergo biologically relevant interactions with nitrogen oxides imply that the cellular concentration of nitrosated and nitrated species may effectively comprise a reserve or cellular store. Potentially, these nitroso reserves function as critical components of the overall redox status of the intracellular environs. Understanding the dynamic regulation of nitric oxide/nitrogen oxides release from these stores is likely to provide clues important in resolving the complex pathophysiology of poorly understood multifactorial disorders, including neurodegeneration, multiorgan failure, cardiomyopathy, and septic shock. PMID- 11396480 TI - Prooxidant and antioxidant functions of nitric oxide in liver toxicity. AB - In response to tissue damage and inflammation induced by a variety of xenobiotics including acetaminophen, carbon tetrachloride, ethanol, galactosamine, and endotoxin, as well as disease states such as viral hepatitis, and postischemic and regenerative injury, the liver produces large quantities of nitric oxide. Indeed, nearly all cell types in the liver including hepatocytes, Kupffer cells, stellate cells, and endothelial cells have the capacity to generate nitric oxide. Thus, these cells, as well as infiltrating leukocytes, may indirectly augment tissue injury. In many models of liver damage, nitric oxide and its oxidation products such as peroxynitrite contribute to the injury process by directly damaging the tissue or by initiating additional immunologic reactions that result in damage. In some models, nitric oxide donors or peroxynitrite can mimic the cytotoxic actions of liver toxins. Moreover, agents that prevent the generation of nitric oxide or antioxidants that bind reactive nitrogen intermediates, or knockout mice with reduced capacity to produce nitric oxide, are protected from xenobiotic-induced tissue injury. In contrast, there have been reports that blocking nitric oxide production enhances xenobiotic-induced tissue injury. This has led to the concept that nitric oxide either inactivates proteins critical for xenobiotic-induced tissue injury or acts as an antioxidant, reducing cellular levels of cytotoxic reactive oxygen intermediates. Whether or not nitric oxide or secondary oxidants generated from nitric oxide act as mediators of tissue injury or protect against toxicity is likely to depend on the precise targets of these reactive nitrogen intermediates, as well as levels of superoxide anion present and the extent to which tissue injury is mediated by reactive oxygen intermediates. In addition, as toxicity is a complex process involving a variety of cell types and many soluble mediators, the contribution of each of these factors must be taken into account when considering the role of nitric oxide as a determinant of tissue injury. PMID- 11396481 TI - Nitric oxide and blood-brain barrier integrity. AB - The blood-brain barrier (BBB) is comprised of the endothelial cells that line the capillaries of the brain. The unique characteristics of this barrier include tight intercellular junctions, a complex glycocalyx, a paucity of pinocytic vesicles, and an absence of fenestra. These properties allow for the selective exchange of substances between the systemic circulation and the extracellular fluid compartment of the brain. It is well established that there are many conditions, including those mediated by nitric oxide (NO), that can lead to an opening of the BBB, eventually leading to vasogenic edema and secondary brain damage. The precise molecular mechanisms mediating NO-induced tissue injury and the breakdown of the BBB are complex and not completely understood. NO is a soluble, easily diffusible gas that is generated by NO synthase. Two of the isoforms of NO synthase are constitutive, calcium-dependent enzymes that modulate many physiological functions, including the regulation of smooth muscle contraction and blood flow. The third isoform is calcium-independent and inducible and can be stimulated by stress, inflammation, and infection. Under these conditions, NO can be generated in large quantities and has detrimental effects on the CNS. NO has been shown to increase permeability of the BBB, allowing substances to enter into the brain passively. This review considers the role of NO and BBB integrity. PMID- 11396482 TI - Redox properties of cytochrome c. AB - The redox properties of cytochromes (cyt) c, a ubiquitous class of heme containing electron transport proteins, have been extensively investigated over the last two decades. The reduction potential (E degrees') is central to the chemistry of cyt c for two main reasons. First, E degrees' influences both the thermodynamic and kinetic aspects of the electron exchange reaction with redox partners. Second, this thermodynamic parameter is remarkably sensitive to changes in the properties of the heme and the protein matrix, and hence can be profitably used for the investigation of the solution chemistry of cyt c. This research area owes much to the exploitation of voltammetric techniques for the determination of E degrees' for metalloproteins, which dates back to the late 1970s. Since then, much effort has been devoted to the comprehension of the molecular factors that control E degrees' in cyt c, which include first coordination sphere effects on the heme iron, the interactions of the heme group with the surrounding polypeptide chain and the solvent, and also include medium effects related to the nature and ionic composition of the solvent, pH, the presence of potential protein ligands, and the temperature. This article provides an overview of the most significant advances made in this field recently. PMID- 11396483 TI - The importance of proteins in defense against oxidation. AB - Free radicals are a normal feature of cellular oxygen metabolism. However, free radical-associated damage is an important factor in many pathological and toxicological processes. For a long time, lipid peroxidation, mediated by oxygen derived free radicals, was probably the most extensively investigated process. From more recent studies, it has become evident that proteins are also the targets of free radicals, and this has important implication for their activity, unfolding, and degradation, as well as in cell functioning. After giving a brief overview of the key role of proteins in the overall antioxidant defense, this review examines their role as targets of oxidation reactions, taking into account the reactivity of amino acid residues and some of their oxidation products. In light of recent data, we then consider the specific role of sulfur-containing amino acids in protein degradation and their possible interplay with the reversal of limited oxidative lesions. The participation of proteins in the overall antioxidant defense is also discussed, specifically the role of metallothionein as an intracellular antioxidant and that of albumin as a circulating antioxidant. PMID- 11396484 TI - Redox reactions of hemoglobin and myoglobin: biological and toxicological implications. AB - Direct cytotoxic effects associated with hemoglobin (Hb) or myoglobin (Mb) have been ascribed to redox reactions (involving either one- or two-electron steps) between the heme group and peroxides. These interactions are the basis of the pseudoperoxidase activity of these hemoproteins and can be cytotoxic when reactive species are formed at relatively high concentrations during inflammation and typically lead to cell death. Peroxides relevant to biological systems include hydrogen peroxide, lipid hydroperoxides, and peroxynitrite. Reactions between Hb/Mb and peroxides form the ferryl oxidation state of the protein, analogous to compounds I and II formed in the catalytic cycle of many peroxidase enzymes. This higher oxidation state of the protein is a potent oxidant capable of promoting oxidative damage to most classes of biological molecules. Free iron, released from Hb, also has the potential to promote oxidative damage via classical "Fenton" chemistry. It has become increasingly evident that Hb/Mb redox reactions or their by-products play a critical role in the pathophysiology of some disease states. This review briefly discusses the reactions of Hb/Mb with biological peroxides, potential cytotoxicity and the impact of these interactions on modulation of cell signaling pathways regulated by these reactive species. Also discussed in this article is the role of heme-protein chemistry in relation to the toxicity of hemoproteins. PMID- 11396485 TI - Is the effect of interleukin-1 on glutathione oxidation in cultured human fibroblasts involved in nuclear factor-kappaB activation? AB - Our understanding of the interleukin-1 (IL-1) signaling molecular mechanisms has recently made considerable progress, with the discovery of the IL-1 receptor associated kinase and the downstream enzymatic cascade that leads to the activation of nuclear factor-kappaB (NF-kappaB). IL-1 signaling and especially NF kappaB activation are thought to be redox-sensitive, even though the precise nature and the molecular targets of the oxidants/antioxidants involved remain largely unknown. Here, we investigated the possible role of cellular oxidized/reduced glutathione (GSSG/GSH) balance in IL-1 signaling. We describe a quantitative method based on capillary electrophoresis designed to assay both intracellular GSH and GSSG in adhering fibroblasts. This method allows the GSSG/GSH balance to be followed during IL-1 stimulation. Our data show that IL-1 induces rapid and transient oxidation of intracellular glutathione in human fibroblasts. Using various antioxidants, including pyrrolidine dithiocarbamate and curcumin, we were unable to show a direct relationship between this IL-1 induced glutathione oxidation and NF-kappaB activation. Of the five antioxidants tested, only curcumin was able to inhibit IkappaBalpha degradation upstream and, hence, NF-kappaB DNA-binding activity and NF-kappaB-dependent expression of IL-6 downstream. PMID- 11396486 TI - Venereal disease and gay men: opening remarks. 1977. PMID- 11396488 TI - Hyperphosphatemic tumor calcinosis. PMID- 11396487 TI - The boxy nasal tip: classification and management based on alar cartilage suturing techniques. AB - The boxy nasal tip is characterized by a broad, rectangular appearance of the tip lobule on basal view. This manifests anatomically as one of three types: type I, which features an increased intercrural angle of divergence (greater than 30 degrees) and normal domal arc (4 mm or less) manifesting as the tip-defining points; type II, which features an increased angulation of the domes of the lower lateral segments of cartilage, creating a widened domal arc (greater than 4 mm) and normal angle of divergence (30 degrees or less); and type III, which features a combination of increased angle of divergence (greater than 30 degrees) and widened crural domal arc (4 mm or greater). In this article, the available techniques for correction of the boxy tip are reviewed and an algorithmic approach for the management of this problem is demonstrated using the open approach to rhinoplasty. Using an individualized algorithmic approach with intraoperative nasal tip analysis and three nasal tip suture reshaping techniques, consistent aesthetic results can be obtained in the correction of the boxy nasal tip. PMID- 11396489 TI - Association between transforming growth factor beta 3 and cleft lip and/or palate in the Japanese population. PMID- 11396490 TI - Treatment of subungual hamartomas in patients with tuberous sclerosis. PMID- 11396491 TI - Anterior cervicoplasty in the male patient. PMID- 11396492 TI - Racial distribution of Dupuytren's disease in Department of Veteran's Affairs patients. PMID- 11396493 TI - Warning about a warning about anatomical breast implants. PMID- 11396494 TI - Dermabrasion compared with laser resurfacing. PMID- 11396495 TI - More about the superiority of Pegasus "zero pressure" mattresses. PMID- 11396496 TI - Asymmetric breast implant levels. PMID- 11396497 TI - Breast implants: some considerations about volume. PMID- 11396498 TI - Ectopic bone formation following temporalis muscle transposition for facial paralysis. PMID- 11396500 TI - 10 ways to save time and money in your operating room. PMID- 11396499 TI - Diagrammatic instructions to nursing staff. PMID- 11396501 TI - Cocaine sniffing immediately after rhinoseptoplasty. PMID- 11396502 TI - The roundblock SMAS treatment. PMID- 11396503 TI - Endoscopic bipolar forceps. PMID- 11396504 TI - Polyglycolic acid conduits for digital nerve reconstruction. PMID- 11396505 TI - Characterization of hairpin-duplex interconversion of DNA using polyacrylamide gel electrophoresis. PMID- 11396506 TI - Effect of phytohaemagglutinin on the surface charge of human and murine lymphocytes. PMID- 11396507 TI - Cell replacement in the human stratum corneum in old age. PMID- 11396508 TI - Resistance to antimicrobial Agents (RAA). Monte Carlo, Monaco. 20-23 October 1999. Proceedings of a meeting. PMID- 11396509 TI - Unusual and memorable. Calcifying aponeurotic fibromata. PMID- 11396510 TI - Unusual but memorable. Multicentric reticulohistiocytosis. PMID- 11396511 TI - Unusual but memorable. Osseous involvement in chronic sarcoidosis. PMID- 11396512 TI - Unusual and memorable. Systematic lupus erythematosus--myositis overlap syndrome. PMID- 11396513 TI - Highlights & trends. The year in review--Part II. PMID- 11396514 TI - Repeated cocaine administration does not alter morphine-induced rotational behavior in nigrally denervated rats. AB - Repeated administration of morphine to rats increases their sensitivity to behavioral effects of morphine as well as to those of psychomotor stimulants, such as cocaine and amphetamine. Conversely, stimulant-induced sensitization to behavioral effects of stimulants often results also in sensitization to behavioral effects of morphine. However, in nigrally lesioned rats, repeated injections of morphine produce sensitization to morphine-induced turning but not to turning induced by cocaine or amphetamine. The present study was performed to determine whether giving repeated cocaine injections to nigrally lesioned rats would produce cross-sensitization to morphine-induced turning. Daily injections of 10 mg/kg cocaine (i.p.) enhanced the turning response to cocaine by day 8, but not the turning response to 3.0 mg/kg morphine (s.c.). The response to morphine increased equally in both cocaine- and saline-treated animals after they had received morphine once. Dose-response curves for morphine (1.0-10 mg/kg) and for cocaine (3.0-30 mg/kg), determined during weeks 3 and 4, were the same in rats receiving daily injections of cocaine or daily injections of saline. Thus, although repeated exposure to cocaine or morphine resulted in sensitization to turning induced by each drug, respectively, there was no cross-sensitization between the two drugs. In contrast to other behaviors, rotational behavior does not seem to exhibit cross-sensitization between morphine and psychomotor stimulants. PMID- 11396515 TI - Effect of serotonin (5-HT)1B receptor ligands on cocaine sensitization in rats. AB - Recent studies have shown that antagonists of serotonin (5-HT)1B receptors attenuate cocaine-induced locomotor hyperactivity, whereas agonists enhance reinforcing and discriminative stimulus effects of the psychostimulant. The present study was designed to determine how 5-HT1B receptor ligands affected the development or the expression phase of sensitization to the cocaine-induced locomotor response in rats. In Experiment 1, rats were treated repeatedly (for 5 days) with cocaine (10 mg/kg) in combination with either saline, GR 127935 (5 HT1B antagonist), CP 94,253 (5-HT1B agonist) or GR 127935 + CP 94,253. On day 10, they received a challenge dose of cocaine (10 mg/kg). In Experiment 2, animals received either saline or cocaine (10 mg/kg) for 5 days, and were then challenged with cocaine (10 mg/kg) in combination with saline, GR 127935, CP 94,253 or GR 127935 + CP 94,253, on day 10. In Experiment 3, rats received either saline, cocaine or CP 94,253 for 5 days; on day 10 they received challenge doses of CP 94,253 or cocaine. In rats treated repeatedly with cocaine, the locomotor hyperactivity induced by a challenge dose of the psychostimulant was about twice as high as that observed after its first administration. The effect evoked by cocaine challenge was further increased in animals treated repeatedly with CP 94,253 + cocaine, but not with GR 127935 + CP 94,253 + cocaine. No difference was observed in the response to cocaine challenge in rats treated repeatedly with cocaine or GR 127935 + cocaine (Experiment 1). In animals treated repeatedly with the psychostimulant, the behavioral response to a challenge dose of cocaine was dose-dependently increased when that drug was combined with CP 94,253, but not with GR 127935 + CP 94,253. No difference was observed in the locomotor response of rats challenged with cocaine or GR 127935 + cocaine (Experiment 2). When rats were treated repeatedly with cocaine, a challenge dose of CP 94,253 produced an about threefold increase in the locomotor effect compared to the animals treated likewise with saline (Experiment 3). Our results indicate that 5-HT1B receptors are involved in neither the development nor the expression of sensitization to cocaine-induced locomotor hyperactivity. On the other hand, they also show that pharmacological activation of 5-HT1B receptors enhances both phases of this phenomenon, and that repeated administration of cocaine leads to an increased functional reactivity of these receptors. PMID- 11396516 TI - Two procedures establishing preference for oral cocaine and lidocaine solutions which do not use an associative history with a reinforcer. AB - Groups of rats were exposed to daily, 3-h, fixed-time 1-min food-pellet delivery sessions, a procedure that produces overdrinking (schedule-induced polydipsia, SIP). Previous research demonstrated that rats drinking a drug or non-drug solution come to prefer that solution to water if the solution had (a) a past association with either a highly acceptable vehicle (e.g. glucose/saccharin), or (b) allowed rats to eschew drinking an unacceptable solution under SIP conditions. The present experiments show that under the solution-avoidance procedure, preference for a concurrent, alternative solution (0.19 mg/ml lidocaine or 0.24 mg/ml cocaine) occurred when a concentrated quinine solution alternative was either abruptly removed or faded. A concentrated cocaine solution, however, was minimally effective in producing a preference for 0.19 mg/ml lidocaine to water when cocaine concentration was faded. Flavor/nutrient conditioning (conditioned reinforcement) and solution-eschewing (avoidance) procedures may throw light on the kinds of historical situations that determine the genesis of stable preferences for drugs and other substances. PMID- 11396517 TI - Sensitivity to the effects of sedative-hypnotics on motor performance: influence of task difficulty and chronic phenobarbital administration. AB - The present investigation examined sensitivity to the effects of various sedative hypnotics on motor performance in rats treated chronically with phenobarbital. Eight rats were trained to walk on a rotorod treadmill at low (8 r.p.m.) and high (24 r.p.m.) rotational speeds. Prior to the chronic regimen, phenobarbital, pentobarbital, amobarbital, diazepam and clonazepam produced dose-dependent impairments in motor performance at both speeds. During chronic treatment with phenobarbital (100 mg/kg/day), tolerance was conferred to the effects of all the drugs examined, as evidenced by rightward shifts in their dose-effect curves. For all drugs, the magnitude of this tolerance was generally consistent across the two speeds. Following a 6-week washout period, during which no drugs were administered, dose-effect curves for each drug shifted back toward their original (i.e. pre-chronic) positions. Under all conditions, the doses required for each drug to impair motor performance at the low speed were higher than those required to impair motor performance at the high speed. These data suggest that sensitivity to the motor-impairing effects of sedative-hypnotics is influenced by the difficulty of the behavioral task, but that task difficulty does not modulate the maximal extent to which tolerance and cross-tolerance are expressed. PMID- 11396518 TI - Effects of the NMDA receptor antagonist, D-CPPene, on sensitization to the operant decrement produced by naloxone in morphine-treated rats. AB - Sensitization to the rate-decreasing effects of opioid antagonists induced by acute pretreatment with opioid agonists has been suggested to reflect initial changes in opioid systems that underlie physical dependence. Glutamate receptors are implicated in the development and expression of opioid dependence, and antagonists acting at the N-methyl-D-aspartate (NMDA) subtype of glutamate receptors have been shown repeatedly to attenuate the severity of opioid withdrawal. The present study evaluated the ability of a competitive NMDA receptor antagonist, D-CPPene (SDZ EAA 494; 3-(2-carboxypiperazin-4-yl)-1 propenyl-1-phosphonic acid), to affect morphine-induced sensitization to naloxone in rats trained to lever-press on a multiple-trial, fixed-ratio 10 schedule of food reinforcement. D-CPPene (0.3-3 mg/kg) was administered either 4 h or 30 min prior to the test session. Morphine (10 mg/kg) or its vehicle was administered 4 h before naloxone challenge (0.3-3 mg/kg). D-CPPene failed to prevent morphine induced potentiation of the naloxone-produced decrement in operant performance. Thus, these results suggest that agonist-induced sensitization to behavioral effects of opioid antagonists may be insensitive to NMDA receptor blockade. PMID- 11396519 TI - NMDA receptor antagonists do not block the development of sensitization of catalepsy, but make its expression state-dependent. AB - Dopamine (DA) receptor blockade induces catalepsy in rats which increases in strength upon retesting. This increase in catalepsy represents a form of sensitization which has been shown to be completely context dependent. Sensitization of catalepsy therefore represents a good model for studying the neurobiological mechanisms underlying the interaction between the cellular effect of a drug (DA-receptor blockade) and the context. This study investigated whether glutamatergic mechanisms are involved in the development of sensitization. Rats were treated with either haloperidol or haloperidol plus an N-methyl-D-aspartate (NMDA) receptor antagonist. Haloperidol consistently induced catalepsy which developed sensitization upon retesting. Co-administration of D-CPPene (5 mg/kg and 10 mg/kg, i.p.), eliprodil (30 mg/kg, i.p.) or Ro 25-6981 (15 mg/kg, i.p.) did not have any effect on sensitization, although all three drugs exerted some anticataleptic effects. When sensitization developed under haloperidol plus NMDA receptor antagonist, the sensitized response was expressed only in the presence of the NMDA receptor antagonist. This strongly suggests that the NMDA receptor antagonists represent contextual stimuli to which catalepsy has been conditioned, and this implies that the expression of sensitization has been rendered state dependent. PMID- 11396520 TI - Motivation for alcohol assessed by multiple variable interval schedule behaviour: effects of reward size and alcohol cues. AB - Altered motivation for drugs of abuse is a central feature of most definitions of drug dependence and the impact of drug-related cues on motivation is of current interest. However, since most studies of cue-reactivity have not used behavioural measures of motivation, their results are often difficult to interpret in motivational terms. In the current paper we describe two experiments in which a behavioural technique, based on multiple variable interval (VI) schedules of reinforcement, was used to study motivation for alcohol in human subjects. In both experiments, subjects attended for several sessions and, during each session, were exposed to a 6-ply VI schedule (values ranged from 1 to 720 s), during which they earned points that were later exchanged for a preferred beer or lager. In Experiment 1 the procedure was validated by showing that changes in the magnitude of reinforcement altered behaviour appropriately. In Experiment 2 we found evidence that the presence of an alcohol-related cue increased the value of alcohol rewards. The results are discussed with reference to a model for the behavioural effects of drug-related cues in triggering relapse and a number of problems we found in using the multiple variable interval schedule procedure. PMID- 11396521 TI - Effects of negative punishment contingencies on cocaine self-administration by rhesus monkeys. AB - Although punishment contingencies are widely used with human drug users, basic research on the effectiveness of these procedures is limited. The present study evaluated the effects of a negative punishment contingency, response-contingent timeout (TO) presentation, on cocaine-maintained responding. Rhesus monkeys were trained under a multiple fixed-interval (FI) 5-min cocaine, conjoint FI 5-min cocaine, variable-interval (VI) 30-sec TO schedule. TO values were either 0 (baseline), 10, 30, or 60s in length. During the TO periods, the FI clock continued to operate but the discriminative stimuli signaling cocaine availability were removed, and responding had no scheduled consequence. Cocaine maintained responding in all monkeys and the dose-effect curve was characterized as an inverted U-shaped function. The response-contingent TO presentations reduced response rates maintained by cocaine in all monkeys compared to baseline. The magnitude of the reduction in response rates was not a function of the length of the TO period (i.e. intensity of the punisher), and the punishment effect was enhanced by increases in cocaine dose. When responding was punished, response rates in the unpunished components either also decreased (i.e. response induction; approximately 30% of the cases) or were not affected (approximately 60%). These results demonstrate that cocaine-maintained behavior can be decreased by environmental manipulations involving negative punishment contingencies. PMID- 11396522 TI - Hepatitis C virus infection: 10 years after the discovery of the virus. AB - The hepatitis C virus (HCV) causes an acute but very often chronic liver disease. An estimated 3% of the world population is chronically infected with HCV. Chronic hepatitis C is the major cause of cirrhosis and hepatocellular carcinoma (HCC), which most often lead to liver transplantation. HCV is a single-stranded enveloped RNA virus; it belongs to the flaviviridae family. The virus has been classified into six genotypes, some of which are distributed worldwide, others of which are confined to more restricted areas. The genotype is an independent predictor of response to antiviral treatment. Blood transfusion was a major risk factor for acquiring HCV infection before donor screening for surrogate marker testing for non-A, non-B (NANB) hepatitis began in the mid-1980s, followed by screening for antibody to HCV in 1990. Today, intravenous drug use and high-risk sexual activity are the most frequently identified risk factors associated with HCV infection. The prevalence of people with unknown HCV infection worldwide is high, so it is necessary to screen people with risk factors. The treatment of patients with chronic HCV infection who have not been treated previously should consist of interferon alpha (IFN-alpha) and ribavirin. PMID- 11396523 TI - What is (cost) effective in patients with chronic hepatitis C virus infection? AB - Hepatitis C virus (HCV) infection often progresses to chronic hepatitis, cirrhosis, and possibly hepatocellular carcinoma. Chronic hepatitis C infection is a leading cause of chronic liver disease and the most common indication for liver transplantation. Combination therapy of interferon alpha and ribavirin is currently the standard regimen for chronic hepatitis C. This combination can achieve viral clearance in approximately 40% of patients, and improve histology and prognosis. The most cost-effective approach to guide duration of combination therapy is HCV genotyping. Cost effectiveness cannot be improved further by taking other well-defined predictive factors for sustained virological response into account. Recent insights into HCV kinetics and the correlation between initial viral decline and sustained virological response will allow us to optimize and individually tailor antiviral treatment Individualized treatment according to the initial viral decline, together with further improvements in drugs (e.g. by long-acting pegylated interferons), will have new impact on antiviral efficacy and cost effectiveness. PMID- 11396524 TI - Prevalence of hepatitis C virus infection in the general population of northern Spain. AB - OBJECTIVES: To estimate the prevalence of hepatitis C in a population of northern Spain and describe (i) the risk factors associated with infection and (ii) the distribution of genotypes. DESIGN: Randomized cross-sectional study. METHODS: A random sample of 1,170 people participated in the study. Sociodemographic data were obtained. Antibodies against hepatitis C virus (anti-HCV) and hepatitis C virus (HCV) genotypes were determined. RESULTS: Nineteen of 1,170 (1.6%) subjects were anti-HCV positive (95% CI 1.0-2.6%). In 12 cases (63%), viraemia was present, and the predominant genotype was 1 b (80%). Anti-HCV positive subjects were older than anti-HCV negative subjects (55.8 +/- 15.3 v. 44.8 +/- 20.9; P = 0.02). Two peaks of maximum frequency were found (in the fourth decade and in those over 60 years). Parenteral drug addiction predominates among those of the fourth decade, while transfusion and surgery predominate in people over 60 years. Three (16%) subjects knew they were carriers of HCV. Only three variables remained significant in the multivariate model (illegal drug use, P< 0.0001; previous hepatitis, P< 0.0001; and age, P< 0.02). CONCLUSIONS: Our study emphasizes the need to develop health policies that can cope with the foreseeable increases in the problems associated with HCV infection in the near future. PMID- 11396525 TI - A pragmatic and cost-effective strategy of a combination therapy of interferon alpha-2b and ribavirin for the treatment of chronic hepatitis C. AB - BACKGROUND: Combination of interferon (IFN) alpha and ribavirin is considered the standard treatment for patients with chronic hepatitis C. While combination therapy is more effective than IFN alone, the optimal management of combination treatment remains uncertain. OBJECTIVE: To assess a pragmatic and cost-effective strategy for the therapy of treatment-naive patients with chronic hepatitis C. DESIGN: Markov model on original data of two randomized trials. METHODS: A validated computer simulation model was applied to non-cirrhotic hepatitis C virus (HCV)-infected patients. Patient characteristics and efficacy of treatment were extracted from two randomized trials reporting on 1,445 non-cirrhotic patients. Different strategies were compared separately for genotype 1 and genotype non-1 (mostly genotype 2/3) infections: (1) no treatment; (2) IFN for 48 weeks (if at 12 weeks HCV RNA undetectable); (3) IFN and ribavirin for 24 weeks; (4) IFN and ribavirin for 48 weeks; (5) IFN and ribavirin for 48 weeks (if at 24 weeks HCV RNA undetectable). All strategies were tested for different combinations of known response factors. RESULTS: In genotype non-1 infection, 24 weeks of combination therapy dominates all other strategies. In genotype 1 infection, 48 weeks of combination therapy for week-24 responders only prolongs life expectancy at a favourable cost-effectiveness ratio (CE) of 7,135 euros per quality-adjusted life year (QALY). Taking response factors other than genotype into account does not add to the effectiveness or cost effectiveness. CONCLUSION: Treating non-cirrhotic patients with chronic hepatitis C according to genotype only is most cost effective independent of the number of other known response factors. PMID- 11396526 TI - Hepatocyte proliferation and cell cycle phase fractions in chronic viral hepatitis C by image analysis method. AB - OBJECTIVE: Chronic hepatitis is characterized by necrosis of liver cells, accompanied by an inflammatory reaction and compensatory cell proliferation. The interaction of the core and non-structural proteins of hepatitis C virus (HCV) with several cellular factors suggests that cell proliferation may be influenced by HCV. The aim of this study was to investigate hepatocyte proliferation and DNA ploidy patterns in patients with chronic viral hepatitis C (CH-C) compared with chronic non-viral hepatitis (CH-N), using a TV image analysis method. METHODS: The DNA index (DI) and cell phase fractions (G1, S, G2) were measured by means of digital picture analysis method on nuclear suspensions of Feulgen stained hepatocytes. Cells were taken from the liver biopsy specimens of 71 patients with CH-C and 24 patients with CH-N. Twenty-six normal liver samples were used as controls. RESULTS: Significantly higher G1 (94 +/- 4) and lower S (3.56 +/- 3.16) phase fractions were measured in CH-C compared with CH-N (G1, 90 +/- 6; S, 6.4 +/ 5.99). The DI of moderate (1.12 +/- 0.05) and severe (1.12 +/- 0.05) CH-C showed near-aneuploid DNA content, while diploidy (DI < 1.10) was detected in cases of CH-N. CONCLUSION: The higher G1 and lower S cell cycle phase fractions in CH-C reflect decreased hepatocyte proliferation compared with CH-N. The near-aneuploid DNA content of the HCV-infected liver samples may be a sign of increased genetic instability, which may contribute to the carcinogenic potential of HCV. PMID- 11396527 TI - Chronic hepatitis C with normal or abnormal aminotransferase levels: is it the same entity? AB - OBJECTIVE: The features of hepatitis C virus (HCV) infection with persistently normal serum alanine aminotransferase (ALT) activity levels are not well defined. This study evaluated the characteristics of HCV infection according to the presence or absence of elevated ALT. METHODS: Demographic data, liver histology and HCV genotype were studied in a group of 80 HCV-RNA-positive subjects with persistently normal ALT (PNALT) (group 1), and compared with a second group of 455 HCV-RNA-positive patients with elevated ALT (group 2). The annual progression of liver fibrosis was also calculated. RESULTS: A higher proportion of women was found in group 1:64% vs 42% in group 2 (P< 0.0002). The HCV genotype 1 was less frequent in group 1:49% vs 60% in group 2 and genotype 2 was more frequent: 16% in group 1 vs 4% in group 2 (P< 0.002). Cirrhosis was less frequent in group 1 (4% vs 13% in group 2 (P< 0.0001)). Normal liver was more frequent in group 1:9% vs 1% in group 2 (P< 0.0001). The Knodell score was significantly different between the two groups: 3.2 +/- 0.27 vs 7.15 +/- 0.22 (P< 0.0001). The progression of liver fibrosis was lower in group 1: 0.053 +/- 0.14 units/year vs 0.13 +/- 0.24 in group 2 (P < 0.007). CONCLUSION: HCV infection with PNALT is associated with less severe histological liver disease and a lower fibrosis progression rate. This suggests that the natural history of HCV infection in these patients is different from that in patients with abnormal ALT. PMID- 11396528 TI - Outcome of liver disease in a large cohort of histologically proven chronic hepatitis C: influence of HCV genotype. AB - OBJECTIVE: To assess the influence of hepatitis C virus (HCV) genotypes on the clinical outcome of liver disease, we analysed 2,307 patients. RESULTS: The most frequently represented genotypes were 1b (40%) and 2 (28.1%). Patients with these genotypes had a median age higher than patients with other genotypes (P< 0.01). The overall survival of subjects with genotype 1b was poorer than the survival of patients with other genotypes (P< 0.01). Liver cirrhosis was found in 280 patients (12.1%), and type 1b was the most represented isolate among them (P< 0.01). Sixty-two patients (22%) developed hepatocellular carcinoma (HCC) during a follow-up of 1481.8 cumulative years (estimated crude incidence rate, 4.1 cases per 100 person-years for all cirrhotics; 5.9 cases for genotype 1a; 4.5 cases for genotype 1b; and 2.8 cases for genotypes non-1). Considering the whole population of 2,307 patients, only genotype 1b was associated significantly with both cirrhosis and the development of HCC. One hundred and nineteen cirrhotic patients underwent treatment with interferon in uncontrolled studies. Interferon therapy was associated with both better survival (P< 0.01) and a lower cumulative hazard for HCC (P< 0.01). CONCLUSIONS: Genotype 1b was associated with a poorer prognosis, probably because it leads to cirrhosis and consequently to HCC development. However, our data did not confirm genotype 1b as an independent risk factor for HCC in liver cirrhosis, which plays a major role in carcinogenesis. Interferon should be considered as a useful strategy in cirrhosis for improvement of survival and reduction of HCC risk. PMID- 11396529 TI - Prevalence of fibromyalgia, anxiety and depression in chronic hepatitis C virus infection: relationship to RT-PCR status and mode of acquisition. AB - BACKGROUND: Musculoskeletal complaints, dry eyes, fatigue and anxiety are common symptoms in patients with hepatitis C virus (HCV) infection, but there are few controlled data evaluating this. AIM: To assess the prevalence of rheumatological disease, fatigue and anxiety in different groups of patients with chronic HCV infection. PATIENTS AND METHODS: Seventy-seven patients with HCV were evaluated. Of these, 49 (64%) had been infected via contaminated anti-D immunoglobulin, 25 (33%) were intravenous drug users (IVDUs), and three were transfusion related; 78% were female. Twenty-five age- and sex-matched controls were also evaluated. Assessment was performed by history, physical examination, the Fibromyalgia Impact Questionnaire (FIQ) and the Hospital Anxiety and Depression Score (HADS). RESULTS: Four (5%) patients fulfilled the criteria for fibromyalgia. All were infected via anti-D immunoglobulin, and three were PCR positive. The mean number of tender points in anti-D patients was 5.0 (+/- 4.07) compared with 2.8 (+/- 2.7) in controls (P= 0.028) and 2.5 (+/- 2.2) in IVDUs (P< 0.004). There was no significant difference in the number of tender points between PCR-positive and PCR-negative patients (P= 0.23). Anxiety and depression scores were significantly higher in anti-D patients (P= 0.0001) and IVDUs (P= 0.005) compared with controls. Forty per cent of the HCV patients had a positive Schirmer test. Forty two per cent of PCR-positive patients had a positive rheumatoid factor (RF, > 1/80). CONCLUSION: This study reveals a moderate increase in prevalence of fibromyalgia in HCV patients. The number of tender points was related to mode of acquisition but not to PCR status. Anxiety and depression levels are also increased in HCV patients compared with controls. Prevalence of RF was higher in PCR-positive patients compared with controls and those who had cleared the virus. PMID- 11396530 TI - Brain electrical activity mapping of EEG for the diagnosis of (sub)clinical hepatic encephalopathy in chronic liver disease. AB - We studied the role of brain electrical activity mapping (BEAM) in the assessment of neuropsychiatric disturbances in 48 cirrhotic patients without clinical evidence of hepatic encephalopathy (no HE, n = 19), with subclinical HE (grade 0, denoting pathological psychometric tests, n = 13) and mild-to-moderate HE (grade I, n = 6; grade II, n = 10). Results were compared with 23 healthy controls. BEAM variables quantified were: (i) the peak frequency (PF); (ii) the amplitude of PF; and (iii) the topographic localization of the maximum peak amplitude digitized for quantification by using a coordinate system. Mean amplitudes and their topographic localization in the following frequency-bands were analysed: delta (1.0-3.5 Hz), theta (4.0-7.5 Hz), alpha 1 (8.0-9.5 Hz), alpha 2 (10.0-11.5 Hz), beta 1 (12.0-15.5 Hz), beta 2 (16.0-19.5 Hz), and beta 3 (20.0-23.5 Hz). The PF was significantly slower in all HE patients than in healthy controls (8.5 +/- 2.0 Hz v. 10.1 +/- 1.0 Hz, P< 0.001). Even in no HE, the PF was significantly slower than in controls (8.6 +/- 1.5 Hz v. 10.1 +/- 1.0 Hz, P< 0.01). No relevant topographic differences of PF were observed. The mean amplitudes of the following bands differed significantly between controls and patients: theta (increased in HE, P< 0.05), alpha 2 (decreased in HE, P< 0.05), and beta 2 and beta 3 (increased in HE, (P < 0.05). In HE patients, the topographic localization of all beta bands showed a significant shift from parieto-occipital areas to central areas of the cortex. We conclude that BEAM is a sensitive tool for detecting neuropsychiatric disturbances in cirrhotics with no HE and with subclinical HE. The combination of PF in the theta band, increased mean amplitude in the beta 2 band, and the localization of the latter band in the frontocentral area of the cortex is an objective and sensitive tool for identifying neuropsychiatric disturbances in 85% of cirrhotic patients with no HE. Further studies are required to determine the clinical implications of these abnormal findings in the absence of overt clinical symptoms. PMID- 11396531 TI - Dual-energy X-ray absorptiometry registers bone mineral in the liver. AB - OBJECTIVE: To analyse the difference in bone mineral content (BMC) between the left and right trunk generally obtained by dual-energy X-ray absorptiometry (DXA) by creating identical images over the liver region and the contralateral side. PATIENTS AND DESIGN: Fifty-four patients were selected at random from 1,722 subjects examined by DXA because of osteoporosis. Another five patients were selected who had been followed for osteoporosis by repeated DXA two to five times at intervals from 2 to 36 months. One healthy volunteer was followed for one day by means of DXA total body measurements. All protocols were analysed with respect to BMC, fat mass (FM) and lean tissue mass (LTM) of the imaged trunk and liver. RESULTS: BMC of the right trunk exceeded that of the left trunk in 78% of the investigated subjects. The right (liver) image dominated in all 81 investigations calculated from 60 subjects. There were intraindividual short- and longterm variations between repeated DXA examinations. The amounts of FM and LTM were distributed symmetrically between the right and left trunk. CONCLUSIONS: DXA registers BMC in the liver, which explains the general dominance of the right trunk. The absorption over the liver region varies in the same individual in repeated measurements at intervals of hours to months. PMID- 11396532 TI - Treatment of refractory hepatic hydrothorax with transjugular intrahepatic portosystemic shunt: long-term results in 40 patients. AB - BACKGROUND/AIMS: Hepatic hydrothorax is a complication of portal hypertension secondary to ascites. In this study, we investigated retrospectively the effects of the transjugular intrahepatic portosystemic shunt (TIPS) on hepatic hydrothorax refractory to diuretic treatment. METHODS: Forty patients (Child-Pugh class B, 24 patients; Child-Pugh class C, 16 patients) with hydrothorax refractory to diuretic treatment, pleurocenteses or pleurodesis were included. The TIPS implantation was successful in all patients, who were then followed for 16 +/- 14 months (range 1 day-54 months). RESULTS: TIPS reduced the portosystemic pressure gradient from 26 +/- 6 to 10 +/- 5 mmHg. In the 17 patients whom we followed for 12 months or longer, improvements were found for the Child--Pugh score (8.6 +/- 1.8 v. 6.7 +/- 1.5), serum albumin concentration (3.1 +/- 0.5 v. 3.6 +/- 0.5 g/l), and urinary sodium excretion (22 +/- 29 v. 89 +/- 43 mmol/24 h) (P< 0.05). Two patients developed severe hepatic encephalopathy requiring shunt occlusion. Hydrothorax improved in 82% of patients and resolved in 71% of patients. Fifty per cent of patients developed shunt insufficiency within 7 +/- 9 months, contributing to a probability of relapse-free 1-year survival of 35%. In these patients, shunt revision resulted in a secondary response rate of 82.3%. The 1-year survival was 64%. Both hydrothorax response and survival showed a significant inverse correlation with age over 60 years (P< 0.01 and P< 0.003, respectively) but not with other biomedical variables. CONCLUSION: TIPS is effective for hydrothorax refractory to diuretic treatment and other standard interventions to bridge the time to transplantation. Patients older than 60 years have a poor response and short survival. PMID- 11396533 TI - Gastro-oesophageal reflux and duodenogastric reflux before and after eradication in Helicobacter pylori gastritis. AB - OBJECTIVE: Helicobacter pylori and duodenogastric reflux (DGR) are both associated with chronic gastritis, peptic ulcer and gastric cancer. The nature of their interrelationship remains unclear. H. pylori eradication has also been reported to result in new or worsening acid gastro-oesophageal reflux (GOR). The aim of this study was to investigate the relationship between GOR, DGR and H. pylori infection. METHOD: 25 patients with H. pylori gastritis underwent ambulatory 24-hour oesophageal and gastric pHmetry and gastric bilirubin monitoring before and 12 weeks after H. pylori eradication, confirmed by 14C urea breath testing (UBT). Ten healthy subjects served as a control group. RESULTS: There were no differences between patient and control groups for gastric alkaline exposure or gastric bilirubin exposure (P> 0.25 in all categories). Oesophageal acid reflux was higher in the study group (P< 0.02). No differences were detected in oesophageal acid reflux, gastric alkaline exposure, or gastric bilirubin exposure (P = 0.35, 0.18 and 0.11, respectively) before and after eradication. CONCLUSIONS: Acid GOR is not increased by H. pylori eradication. DGR in patients with H. pylori gastritis is similar to that in healthy, non-infected subjects. H. pylori eradication produces no change in GOR or DGR. In patients with chronic gastritis, H. pylori infection and DGR appear to be independent of each other. PMID- 11396534 TI - Young dyspeptic patients: with a test-and-treat policy, are the benefits of decreased symptom severity and oesophago-gastric-duodenoscopy workload sustained? AB - OBJECTIVES: To examine the symptom severity and requirement for oesophago-gastro duodenoscopy (OGD) in young dyspeptic patients 2 years after serological testing for Helicobacter pylori, and to compare the outcomes of seronegative patients with those of seropositive patients. DESIGN: Long-term follow-up study of 232 participants from our previous trial. METHODS: Telephone assessment of patients' symptom severity, scored using a previously validated questionnaire; return of patients for OGD determined using local patient administration system (PAS); and review of medical case notes. RESULTS: Dyspepsia symptom severity of both seronegative and seropositive patients remained reduced compared with initial scores at time of trial recruitment. Symptom severity of seropositive patients was significantly lower than that of seronegative patients (P< 0.001). Seventeen additional patients returned for OGD between six months and two years after the start of the serological study. As 61 of the original 232 study patients returned for OGD within the first six month follow-up period, a total of 78 individuals (34%) had OGD during the two years following the study. Thus, 66% of the original participants avoided OGD. CONCLUSIONS: This study of non-invasive testing for H. pylori in young dyspeptic patients has demonstrated an improvement in symptom severity and a substantial reduction in OGD workload over a 2-year period. PMID- 11396535 TI - Ranitidine bismuth citrate-based triple therapy for seven days, with or without further anti-secretory therapy, is highly effective in patients with duodenal ulcer and Helicobacter pylori infection. AB - OBJECTIVE: To compare the efficacy of two protocols for the eradication of Helicobacter pylori infection and the healing of active duodenal ulcer: (i) ranitidine bismuth citrate (RBC) plus two antibiotics for 7 days, and (ii) the same triple therapy followed by 3 weeks of anti-secretory drug treatment. METHODS: The study comprised 102 patients with active duodenal ulcer and H. pylori infection; the patients were randomized to open treatment with either RBC 400 mg b.d. plus amoxycillin 1 g b.d. and clarithromycin 500 mg b.d. for 7 days, or the same treatment followed by 3 weeks of RBC 400 mg b.d. alone. Ulcer healing was confirmed by endoscopy. H. pylori eradication was assessed by endoscopy, rapid urease test and histology. RESULTS: The ulcer healed in 48/50 patients on RBC-based triple therapy alone (96.0%) and in 51/52 patients on triple therapy plus further anti-secretory treatment (98.1%). On an intention-to-treat basis, H. pylori had been successfully eradicated in 42/50 patients on triple therapy (84.0%) and in 44/52 patients on triple therapy plus anti-secretory treatment (84.6%), while by per protocol analysis the H. pylori eradication rates were 91.3% (42/46) and 89.8% (44/49), respectively. CONCLUSIONS: One-week triple therapy with RBC, amoxycillin and clarithromycin is highly effective in eradicating H. pylori and healing duodenal ulcers, even if not followed by anti secretory drug treatment. PMID- 11396536 TI - Interleukin-1, interleukin-8, tumour necrosis factor alpha and interferon gamma stimulate DNA synthesis but have no effect on apoptosis in small-intestinal cell lines. AB - OBJECTIVES: Cytokines stimulate lymphocyte cell proliferation and affect cell division in several other cell types. Helicobacter pylori-induced gastritis and coeliac disease are characterized by an increased cell proliferation in association with an increased production of proinflammatory cytokines, which could contribute to these cell kinetic changes. Our aim is to examine in vitro whether cytokines usually present in the gastrointestinal mucosa affect DNA synthesis and apoptosis in a rat and a human small-intestinal cell line. METHODS: IEC-6 and FHs-74 cells were incubated for 24 h with 10(-13)-10(-9) M of tumour necrosis factor alpha (TNF-alpha), interleukin-1beta (IL-1beta), interleukin-2 (IL-2), interleukin-6 (IL-6), interleukin-8 (IL-8), transforming growth factor beta (TGF-beta) and interferon gamma (IFN-gamma). IEC-6 cells were also incubated with 10(-13)-10(-9) M of interleukin-1alpha (IL-1alpha) and 10(-8) M of interleukin-1 receptor antagonist (IL-1ra). The cells were labelled with 3H methyl thymidine for the final 4 hours, and then processed for autoradiography. DNA synthesis was evaluated by the labelling index (LI%). Apoptosis was evaluated in IEC-6 cells by changes in membrane lipid asymmetry using annexin-V binding to externalized phosphatidylserine (flow cytometry) and by estimating the caspase activity. RESULTS: TNF-alpha, IL-1beta, IL-8 and IFN-gamma significantly and markedly increased the LI, even at low concentrations (P< 0.0001), in both IEC-6 and FHs-74 cells, as did IL-1alpha in IEC-6 cells. TGF-beta significantly reduced the LI in both cell lines (P< 0.0001), whereas IL-2, IL-6 and IL-1ra did not affect DNA synthesis significantly. None of IL-1beta, IL-8, TNF-alpha or IFN gamma affected apoptosis in IEC-6 cells. CONCLUSION: TNF-alpha, IL-1alpha, IL 1beta, IL-8 and IFN-gamma stimulated DNA synthesis in a human and a rat small intestinal cell line. The cytokines exert their mitogenic action directly on the intestinal cells via specific receptors. Our findings indicate that pro inflammatory cytokines may participate in the regulation of the gastrointestinal epithelial cell proliferation in health and disease. PMID- 11396537 TI - Defining gluten refractory enteropathy. AB - OBJECTIVE: The diagnosis of coeliac disease (CD) is based on the responsiveness of the enteropathy to a gluten-free diet (GFD). This implies that terms such as 'non-responsive CD' and 'refractory CD' are almost paradoxical. In spite of this, these terms are commonly used in the literature, often with different and confusing meanings. METHODS: On the basis of both a review of the literature and our clinical experience, we propose the following classification. A condition characterized by a refractory enteropathy, not due to lymphoma, ulcerative jejunoileitis or collagenous sprue, but in which gluten sensitivity has been shown previously or could be shown while the patients were on an immunosuppressive therapy should be indicated as refractory CD. Those patients in whom gluten sensitivity can be excluded should be considered to be affected by non-coeliac refractory sprue. Finally, patients in whom the presence of CD cannot be either confirmed or excluded should be considered to be affected by undefined sprue. RESULTS: Twenty-four certain refractory patients are described in the literature. The data suggest a diagnosis of refractory CD in 13 patients, non coeliac refractory sprue in three patients, and undefined sprue in eight patients. CONCLUSIONS: We define refractory CD as a form of CD that no longer responds to a GFD. Non-coeliac refractory sprue is a condition unrelated to CD. It could be either an independent condition or a common end point of different enteropathies. PMID- 11396538 TI - Influence of inflammatory bowel disease on different dimensions of quality of life. AB - OBJECTIVE: To establish the impairment of different dimensions of quality of life in inflammatory bowel disease (IBD). DESIGN: Prospective observational study. PARTICIPANTS: 289 patients [160 with ulcerative colitis (UC) and 129 with Crohn's disease (CD)]. MEASURES: Health-related quality of life was assessed by means of the Inflammatory Bowel Disease Questionnaire (IBDQ) and the Psychological General Well Being Index (PGWBI). RESULTS: In active IBD, all dimensions of the quality of life scored significantly lower than in inactive IBD, indicating a poor quality of life. Social impairment was the least impaired dimension of the IBDQ in active UC and CD, compared with digestive and systemic symptoms. In inactive IBD, the systemic symptoms domain received the lowest score (P < 0.01). In a subgroup of 22 patients studied before and after remission, emotional function was the most impaired dimension after achieving remission. The Psychological General Well Being Index was significantly impaired in active UC [78.5 (range 64 89)] and CD [76.5 (range 69-97)] relative inactive IBD [104 (range 93-111)] vs 106 (95-113), respectively; P < 0.05]. CONCLUSIONS: Quality of life is impaired in IBD. During relapse, clinicians should pay attention to digestive symptoms and psychological distress. In remission, they should be sensitive to systemic symptoms. PMID- 11396539 TI - Intrarectal injection of glycerol induces hypersensitivity to rectal distension in healthy subjects without modifying rectal compliance. AB - BACKGROUND: Rectal sensory thresholds are lowered in patients with irritable bowel syndrome (IBS), reflecting visceral hyperlagesia, which might be related to subclinical inflammation. AIM: To evaluate the effects of an intraluminal injection of glycerol, a mucosal irritant, on rectal tone and perception of distension in 12 healthy subjects. METHODS: Rectal tone was evaluated with a barostat. First sensation, need to defecate and pain thresholds were evaluated during isobaric phasic distensions, before and 20 and 120 min after injection of 10 ml glycerol in the rectum. RESULTS: Baseline bag volume (97.9 +/- 56.2 ml) significantly decreased 20 min (49.7 +/- 42.2 ml; P= 0.026) and 120 min (66.5 +/- 38.3 ml; P= 0.050) after injection of glycerol, indicating its hypertonic effect. The pressure defining sensory thresholds was decreased significantly 20 min after glycerol injection: first sensation, 14.6 +/- 2.9 versus 18.3 +/- 7.2 mm Hg (P = 0.01); need to defecate, 19.6 +/- 3.7 versus 26.0 +/- 6.9 mm Hg; pain, 23.8 +/- 4.5 versus 35.6 +/- 9.5 mm Hg (P = 0.001). This effect was maintained for 120 min after injection of glycerol. Slopes of the compliance curves did not differ before and after injection of glycerol. CONCLUSIONS: Intraluminal injection of glycerol significantly increases rectal tone and sensitizes healthy volunteers to rectal distension, since they show significantly lower thresholds after glycerol. This could constitute a model of visceral hypersensitivity in healthy volunteers. PMID- 11396540 TI - Tuberculous peritonitis--reports of 26 cases, detailing diagnostic and therapeutic problems. AB - OBJECTIVE: To evaluate the clinical presentation, biochemical (ascites and serum) and laparoscopic findings, and to assess the efficacy of triple antituberculous therapy without rifampicin for 6 months in patients with tuberculous peritonitis. METHODS: Twenty-six tuberculous peritonitis patients (11 male, 15 female) with a mean age of 34.8 +/- 3.4 years (range 14-77) were assessed with regard to diagnostic and therapeutic features. RESULTS: The most common symptoms and signs were abdominal pain (92.3%) and ascites (96.2%), respectively. Tuberculin skin test (TST) was positive in all patients. An abnormal chest radiography suggestive of previous tuberculosis was present in five patients (19.2%), and two patients (7.7%) had extra-peritoneal (cerebral, pericardial) active tuberculous involvement. In 24 of the 25 patients who underwent laparoscopy with directed biopsy, whitish nodules suggested tuberculous peritonitis; 76% of the biopsy specimens revealed caseating, 20% non-caseating granulomatous inflammation, and 4% non-specific findings. The ascitic fluid of one patient (3.8%) was positive for acid-resistant bacilli, and culture was positive in two patients (7.7%). Twenty-four of the patients were treated for 6 months with isoniazid, streptomycin (total dose 40 g) and pyrazinamide (for the first 2 months and then substituted with ethambutol). Eighteen patients also received methyl prednisolone, initially 20 mg/day, for 1 month. The follow-up period was 19 +/- 1.7 months after the end of therapy (range 6-36). Ascites and abdominal pain abated earlier in patients on steroid therapy. All but two of the 24 patients responded to treatment. CONCLUSION: Non-invasive tests such as acid-fast stain and culture of the ascitic fluid are usually insufficient, hence invasive laparoscopy and peritoneal biopsy are necessary for the diagnosis of tuberculous peritonitis if non-invasive tests such as ascites adenosine deaminase activity measurement are not easily available. Triple therapy without rifampicin for 6 months is sufficient to treat tuberculous peritonitis. PMID- 11396541 TI - Salmonella dublin infection: a rare cause of spontaneous bacterial peritonitis and chronic colitis in alcoholic liver cirrhosis. AB - Salmonella dublin is a veterinary pathogen which rarely causes human illness, although reported human isolates have increased over the past two decades. This serovar of salmonella is unusually invasive and life-threatening, although the clinical pattern of human infection is not well known. We describe a 51-year-old cirrhotic patient who presented with severe liver failure, chronic diarrhoea and left-sided segmental colitis. Radiological and endoscopic findings suggested Crohn's colitis. During the hospital stay he developed a spontaneous bacterial peritonitis (SBP) and S. dublin was isolated in the ascitic fluid. Our report supports the view that this salmonella serovar should be kept in mind as a rare cause of SBP in cirrhotic patients, especially in those cases with chronic colitis resembling Crohn's disease. PMID- 11396542 TI - Autoimmune hepatitis associated with the antiphospholipid syndrome. AB - The antiphospholipid syndrome has rarely been described in patients with autoimmune hepatitis. Two cases with type I autoimmune hepatitis and antiphospholipid syndrome are presented. The first case is that of a 53-year-old Caucasian female with a history of arterial thrombosis and fetal loss who was submitted to clinical and laboratory testing due to persistent transaminasaemia and was found to have autoimmune hepatitis. Antiphospholipid antibodies (anticardiolipin antibodies and lupus anticoagulant) were positive. The second case is that of a 31-year-old Caucasian woman with a history of autoimmune hepatitis who was submitted to laboratory testing due to a second-trimester fetal death, revealing an increased activated partial thromboplastin time and positive antiphospholipid antibodies. In conclusion, secondary antiphospholipid syndrome may accompany autoimmune hepatitis. PMID- 11396543 TI - Necrotizing enterocolitis in a married couple due to a staphylococcal toxin. AB - A married couple affected by necrotizing enterocolitis is described. Both had the same toxin-producing Staphylococcus aureus which was considered to be responsible for the necrotizing enterocolitis. PMID- 11396544 TI - Surreptitious abuse of magnesium laxatives as a cause of chronic diarrhoea. AB - Surreptitious laxative abuse is increasingly considered as a possible cause of unexplained chronic diarrhoea. Laboratory services for detection of colonic stimulant laxatives are widely available; however, laboratory facilities for identifying patients with magnesium-induced diarrhoea are not commonly provided, making diagnosis difficult. We describe three patients who surreptitiously abused magnesium laxatives, and whose diagnoses were delayed, leading in each case to extensive investigations and lengthy in-patient stays. In all three cases, the diagnosis was eventually made by the simple measurement of magnesium in a random faecal sample. We would like to increase the awareness of surreptitious magnesium laxative abuse as a cause of chronic diarrhoea so that costly and unnecessary investigations may be minimized. PMID- 11396545 TI - Indications and efficacy of botulinum toxin in disorders of the gastrointestinal tract. AB - In recent years, botulinum toxin type A (BT) has been found to be effective in the treatment of various spastic disorders of smooth muscle in the upper and lower gastrointestinal tract. The short-term efficacy of intrasphincteric injection of BT in achalasia is now well established, however, because of the chronic nature of the disease, patients will require repeated injections at regular intervals. In contrast, after a single injection into the anal sphincter, BT has impressively high healing rate with minimal side effects. BT remains a novel therapeutic approach in a range of other gastrointestinal motility disorders including diffuse oesophageal spasm, sphincter of Oddi dysfunction and anismus, and the list of its indications is increasing. BT seems to be safe but as its long-term effects remain unestablished, it should be used with caution in younger patients. In this review we discuss the mechanism of action, indications, efficacy and side-effects of BT with its use in various areas of gastrointestinal tract. PMID- 11396546 TI - Proton pump inhibitors and their drug interactions: an evidence-based approach. AB - The proton pump inhibitors (PPIs) are the most effective antisecretory agents used to treat acid-related disorders. As such, they are frequently prescribed for patients who are concurrently using other medications. PPIs may interact with other drugs through numerous mechanisms. The most important include competitive inhibition of hepatic cytochrome P (CYP) 450 enzymes involved in drug metabolism, and alteration of the absorption of other drugs via changes in gastric pH levels. Poor metabolizers, who lack CYP2C19, may be particularly predisposed to drug interactions. Although the potential for drug interactions is high, few clinically significant interactions have been reported for the PPIs. Nevertheless, caution is indicated when certain drugs are co-prescribed with these agents. The incidence of clinically significant drug interactions increases proportionately with the number of drugs taken and with the age of the patient. The drug interaction with the greatest clinical importance is the reduction in benzodiazepine clearance by omeprazole. PMID- 11396547 TI - Neuropsychological profiles of adults with Klinefelter syndrome. AB - Children and adolescents with Klinefelter syndrome (XXY) have been reported to show deficits in language processing including VIQ < PIQ and a learning disability in reading and spelling. However, whether this is characteristic of adults with Klinefelter syndrome has not been established. Thirty-five men with Klinefelter syndrome, aged 16 to 61, and 22 controls were evaluated with a comprehensive neuropsychological battery. The Klinefelter patients scored significantly below controls in language skills, verbal processing speed, verbal and nonverbal executive abilities, and motor dexterity. Within the Klinefelter sample, three cognitive subgroups were identified: VIQ 7 or more points below PIQ (n = 10), VIQ within 6 points of PIQ (n = 12), and PIQ 7 or more points below VIQ (n = 12). The deficits detected in language, verbal processing speed, and verbal executive skills were found to be isolated to the VIQ < PIQ subgroup, while the abnormalities in motor dexterity and nonverbal executive skills were confined to the PIQ < VIQ subgroup. Older age was significantly correlated with increases in VIQ relative to PIQ in the patient group, which suggests the intriguing possibility that the PIQ < VIQ subgroup primarily emerges in young adulthood, perhaps in response to the reported hormonal abnormalities detected in Klinefelter syndrome patients during puberty. PMID- 11396548 TI - Use of the Disability Rating Scale Recovery curve as a predictor of psychosocial outcome following closed-head injury. AB - Rapid rate of recovery has been associated with better outcome following closed head injuries, but few studies have compellingly demonstrated this. This study used growth curve analyses of Disability Rating Scale (DRS) scores at acute hospitalization discharge, 1, 3, and 6 months post injury in a sample of 55 patients with a closed-head injury. Six month post-injury outcome measures were taken from significant other (SO) responses on the NYU Head Injury Family Interview (NYU-HIFI) including severity and burden ratings of affective/neurobehavioral disturbance, cognitive deficits, and physical/dependency status. Rate of recovery (linear and curvilinear recovery curve components) was significantly related to the level of affective/neurobehavioral severity, and the severity and burden of SO-perceived cognitive deficits. Only the intercept of the DRS recovery curve was associated with the SO-perceived severity and burden of physical/dependency status. Growth curve modeling is a meaningful and powerful tool in predicting head injury outcome. PMID- 11396549 TI - Neuropsychological performance of right- and left-frontotemporal dementia compared to Alzheimer's disease. AB - The performance of 16 patients with Alzheimer's disease (AD) was compared to 11 patients with right-frontotemporal dementia (FTD) and 11 patients with left-FTD on a comprehensive neuropsychological battery. Standardized scores (i.e., z scores based on normal control data) were analyzed for 5 cognitive domains. The results revealed that the AD group displayed significant impairment in visual constructional ability relative to the two FTD groups; however, no significant difference was found between the groups on memory scores (verbal and nonverbal). Patients with left-FTD scored significantly below patients with AD on the language measures (e.g., word retrieval, verbal semantic memory), and verbal executive ability (phonemic fluency); AD patients did not differ from patients with right-FTD on these measures. Patients with right-FTD exhibited significantly more perseverative behavior than AD patients; AD patients did not differ from left-FTD patients on this parameter. These results indicate that the pattern of neuropsychological performance of AD patients is distinguishable from patients with left and right frontal frontotemporal dementia. PMID- 11396550 TI - Effects of retroactive and proactive interference on word list recall in schizophrenia. AB - Schizophrenia spectrum patients (N = 143) and healthy controls (N = 160) were administered the Rey Auditory Verbal Learning Test (RAVLT) and tests of executive functioning to directly investigate the effects of proactive interference (PI) and retroactive interference (RI) on word list recall. It was hypothesized that by virtue of the predicted preferential association between executive functioning and RI (relative to PI), patients would demonstrate increased susceptibility to RI in their ability to recall word lists. Results indicated that patients show increased susceptibility to RI relative to PI. Furthermore, this difference appeared to be related to the frontally-mediated central executive functions that were preferentially associated with RI but not PI susceptibility. PMID- 11396551 TI - Abnormally increased semantic priming in children with symptomatic HIV-1 disease: evidence for impaired development of semantics? AB - Language deficits are a major characteristic of neurobehavioral dysfunction in pediatric HIV disease. An object decision task, which assessed reaction time facilitation following a semantic or identical prime in comparison to an unrelated prime, was used to investigate whether semantic processing abnormalities could be responsible, in part, for these deficits. Thirty children with vertically acquired HIV infection (M age 9.0 years; range 6-13) participated. Either a picture of the same object (repetition prime), a semantically related object (semantic prime), a semantically unrelated object, or a nonsense object preceded a target picture, which in 50% of the cases was a real object. Brain scans of children were rated and used together with neurobehavioral functioning to classify children as having HIV-related CNS abnormalities (n = 13) or not (n = 17). Increased semantic priming but not repetition priming was associated with a greater degree of cortical atrophy. Furthermore, CNS compromised children had significantly faster reaction times following a semantic prime compared to an unrelated prime than non-compromised patients. This facilitation following semantic priming for the CNS compromised patients (13.3%) almost equaled the facilitation following repetition priming (15.3%) while for the non-compromised patients facilitation following semantic priming (7.9%) was clearly smaller than following repetition priming (14.6%). These data suggest that HIV infection in children may result in a reduced neural network leading to impoverished semantic representations characterized by poor differentiation between closely related objects. PMID- 11396552 TI - Performance of elderly African American and White community residents on the CERAD Neuropsychological Battery. AB - The CERAD Neuropsychological Battery, includes 7 measures: Verbal Fluency; Modified Boston Naming; Mini-Mental State: Word List Learning, Recall and Recognition; Constructional Praxis. It was originally developed to evaluate patients with a clinical diagnosis of Alzheimer's disease, but is increasingly used in epidemiological studies of the incidence and prevalence of dementia in the elderly. The current study reports norms for African American and White representative community residents 71 years of age and older in North Carolina, and compares performance with that of African Americans in Indianapolis and with Whites in the Monongahela Valley, Pennsylvania. For all 3 studies, increased education and younger age was related to better performance on each of the 7 measures. Sex differences, when present, tended to favor women. Although on average African Americans performed more poorly than Whites, with demographic characteristics controlled, no significant racial differences were found in the North Carolina sample. Both African American and White participants in North Carolina performed more poorly than their racial counterparts in the other 2 studies, possibly because of selection-induced differences in health and educational status. Nevertheless, the use of an identical evaluation battery, such as the CERAD neuropsychologic instrument, facilitates comparisons not otherwise possible, and should be encouraged. PMID- 11396553 TI - Neuropsychological test performance in Aruaco Indians: an exploratory study. AB - A sample of 20 right-handed Aruaco Indians (12 male, 8 female; age 8-30 years) from the Sierra Nevada de Santa Marta (Colombia) participated in this study. A brief neuropsychological test battery (visuoconstructive and visuoperceptual abilities, memory, ideomotor praxis, verbal fluency, spatial abilities, concept formation) was individually administered. In addition, a handedness questionnaire was included. In some neuropsychological tests performance was virtually perfect (Recognition of Overlapped Figures and Ideomotor Praxis Ability test), whereas performance in other tests was impossible (e.g., Block Design using a time limit). It was proposed that two types of variables were significantly affecting performance: (1) educational level; and (2) cultural relevance. Some tests appeared significant and meaningful whereas others were meaningless and even impossible to understand. The appropriateness of current neuropsychological instruments for cross-cultural assessment is discussed. PMID- 11396554 TI - Understanding metaphors and idioms: a single-case neuropsychological study in a person with Down syndrome. AB - The ability of subject F.F., diagnosed with Down syndrome, to appreciate nonliteral (interpreting metaphors and idioms) and literal (vocabulary knowledge, including highly specific and unusual items) aspects of language was investigated. F.F. was impaired in understanding both metaphors and idioms, while her phonological, syntactic and lexical-semantic skills were largely preserved. By contrast, some aspects of F.F.'s executive functions and many visuospatial abilities were defective. The suggestion is made that the interpretation of metaphors and idioms is largely independent of that of literal language, preserved in F.F., and that some executive aspects of working memory and visuospatial and imagery processes may play a role. PMID- 11396555 TI - The effect of exposure to 35,000 ft on incidence of altitude decompression sickness. AB - INTRODUCTION: Exposure to 35,000 ft without preoxygenation (breathing 100% oxygen prior to decompression) can result in severe decompression sickness (DCS). Exercise while decompressed increases the incidence and severity of symptoms. Clarification of the level of activity vs. time to symptom onset is needed to refine recommendations for current operations requiring 35,000-ft exposures. Currently, the U.S. Air Force limits these operations to 30 min following 75 min of preoxygenation. The objective of this study was to determine the effect of exercise intensity on DCS incidence and severity at 35,000 ft. METHODS: Following 75 or 90 min of ground-level preoxygenation, 54 male and 38 female subjects were exposed to 35,000 ft for 3 h while performing strenuous exercise, mild exercise, or seated rest. The subjects were monitored for venous gas emboli (VGE) with an echo-imaging system and observed for signs and symptoms of DCS. RESULTS: Exposures involving strenuous and mild exercise resulted in higher incidence (p < 0.05) and earlier onset of symptoms (p < 0.05) of DCS than exposure at rest. Mild and strenuous exercise during exposure did not differ in incidence or rate of onset. Incidence at 30 min of exposure was 8% at rest and 23% while exercising. CONCLUSION: The results showed that current guidelines for 35,000-ft exposures keep DCS risk below 10% at rest. Exercise, even at mild levels, greatly increases the incidence and rate of onset of DCS. PMID- 11396556 TI - A potential role for free radical-mediated skeletal muscle soreness in the pathophysiology of acute mountain sickness. AB - BACKGROUND: It has been suggested that free radicals may be implicated in the pathophysiology of acute mountain sickness (AMS) due to their ability to initiate and propagate cell membrane damage (3). Therefore, the present study was designed to: a) investigate the effects of an expedition to high altitude on metabolic indices of free radical-mediated oxidative stress and assess subsequent implications for skeletal/cardiac muscle damage; and b) determine whether these parameters were different in subjects who developed AMS after gradual ascent to 5100 m (base camp, BC) compared with those who remained healthy. METHODS: There were 19 male volunteers who were examined at rest and after a standardized maximal exercise test at sea level before and after an expedition (SL1/SL2) and during the first morning of arrival at BC. The trek to BC lasted 20+/-5 d. RESULTS: A mild increase in the Lake Louise AMS score was observed by the end of day 1 at BC (p < 0.05 vs. SL1/SL2). Four subjects developed AMS, which in one subject later progressed to high altitude pulmonary and cerebral edema. The serum concentration of lipid hydroperoxides (LH) increased markedly at rest and after maximal exercise at BC (p < 0.05 vs. SL1/SL2) whereas no changes were observed for plasma malondialdehyde (MDA). Resting serum total phosphocreatine kinase activity (CPK) and myoglobin also increased at BC (p < 0.05 vs. SL1/SL2) whereas cardiac troponin I (cTnI) remained stable. The resting pain threshold decreased and exercise-induced muscle soreness subsequently increased at BC (p < 0.05 vs. SL1/SL2). An association was observed between resting LH and myoglobin at BC (r = 0.45, p < 0.05) and the increase in LH was related to the increase in exercise induced muscle soreness at BC (r = 0.96, p < 0.05). Further correlations were identified between the AMS score on day 1 at BC and: a) resting/exercise LH (r = 0.63, p < 0.05/r = 0.51, p < 0.05); and b) resting pain threshold at BC (r = 0.58, p < 0.05). Furthermore, subjects with AMS on day 1 at BC were characterized by a greater decrease in the resting pain threshold and greater increase in resting LH, CPK and myoglobin compared with subjects without AMS (p < 0.05). Headache, fatigue, insomnia and general apathy were the most frequently reported symptoms of AMS. CONCLUSIONS: Localized free radical-mediated vascular damage of the blood-brain barrier in addition to systemic tissue damage causing overt skeletal muscle soreness may have contributed to the pathophysiology of AMS, the latter through its indirect effects on other non-specific constitutional symptoms such as fatigue and insomnia causing a deterioration in physical performance. PMID- 11396557 TI - Plasma sodium-osmotic dissociation and hormonal interaction with drinking-induced hypervolemia at 2800 m altitude. AB - PURPOSE: To study hormonal factors that may account for the dissociation between beverage-induced plasma sodium p[Na+] and osmotic p[Osm] concentrations that appear to refute the high theoretical correlation between p[Na+] and p[Osm]. METHODS: Ten men (24 +/- SD 3 yr of age) sat reclining (head up) for 12 h in a chamber (21-23 degrees C dry bulb, 25-33% relative humidity) at 2800 m (9184 ft, 539 mm Hg) altitude (ALT), and at 321 m (1053 ft, 732 mm Hg) on the ground (GND). During 1000-1030 hours they consumed 3 fluids (12 ml x kg(-1),X = 948 ml x d(-1)) with large differences in sodium and osmotic contents: AstroAde (AA) with 185 mEq x L(-1) Na+ and 283 mOsm x kg(-1), Performance 1 (Shaklee) (P1) with 22 mEq x L( 1) Na+ and 365 mOsm kg(-1), or H2O at ALT; and only H2O on the GND. RESULTS: After drinking: plasma volume (PV) increased at 1200 hours by 8.3% (p < 0.05) with AA but was not significantly (NS) changed in the other sessions (Xdelta = +0.9%, range -0.9 to 2.8%); p[Na+] and p[Osm] were unchanged. Urinary rates and free-water clearances were attenuated with AA and P1 vs. those with H2O. Correlations between and among p[Na+] and p[Osm] suggest that the pNa+ ion is more tightly controlled than pOsm; and that there was no clear hormonal response that could account for this dissociation from theoretical considerations. CONCLUSIONS: There is significant dissociation between plasma sodium and osmotic concentrations after fluid intake. Induction and maintenance of hypervolemia requires increased (near isotonic) drink Na+ osmols rather than increased non ionic osmols. PMID- 11396558 TI - Light adaptation: night vision goggle effect on cockpit instrument reading time. AB - BACKGROUND: Light adaptation to the intensified image provided by a night vision device may handicap pilots who have set cockpit instrument luminance too low. METHODS: Under conditions simulating night flying, subjects adapted to an NVG image at 3 or 10 footlamberts (fL), then used a joystick to indicate the position of the horizon in an ADI illuminated by NVIS-compatible light at luminances 2 to 3.5 log units lower than the NVG image. RESULTS: Response times increased no more than a few tenths of a second when the decrease in luminance was only 2 log units. Greater decreases produced correspondingly longer delays in response, reaching as much as 5.5 s for subjects in their twenties and 8-15 s for older subjects. CONCLUSIONS: While a decrease of more than 2 log units is not likely to occur under most operational conditions, it is certainly possible, and pilots should be aware that significant risk can be incurred by setting cockpit instruments to luminance levels below 0.03 fL. PMID- 11396559 TI - Cardiac arrhythmias in F-16 pilots during aerial combat maneuvers (ACMS): a descriptive study focused on G-level acceleration. AB - INTRODUCTION: It is well known that acceleration during centrifuge training provokes cardiac arrhythmias. However, there is little research regarding cardiac arrhythmia during actual flight. Therefore, to identify the kinds of cardiac arrhythmias that occur in response to the Aerial Combat Maneuvers (ACM) environment, we analyzed electrocardiograms (ECG) of 35 male pilots. METHODS: ECGs were recorded from 35 pilots (mean age 31 yr, range 27-39) from 1 h before take-off to 1 h after landing. G-load was simultaneously recorded. To see the patterns of the arrhythmia as Gz force increased, ECG data of each subject were classified into three G levels: a) G-level A was < +3 G; b) G-level B was > or = +3 G but < +6 G lasting longer than 3 s; and c) G-level C was > or = +6 G lasting longer than 3 s. RESULTS: Mean total flying time of the 35 pilots was 1475 h (range 578-3300). Arrhythmias were observed in 17 of the pilots (49%) during ECG recording. Four pilots (11.4%) experienced arrhythmias before take-off, 16 pilots (45.7%) inflight and 4 pilots (11.4%) after landing. Arrhythmias were recorded in 13 of 35 pilots (37.1%) at G-level A, in 10 of 35 pilots (28.6%) at G-level B and in 1 of 21 pilots (5%) at G-level C. During flight, unifocal premature ventricular contractions (PVC) were experienced by three of the subjects (8.6%); supraventricular premature beats (SVPB) by four subjects (11.4%), and PVCs with bigeminy by one subject (2.9%). Eight subjects (22.9%) experienced combined arrhythmias. CONCLUSION: There were no clinically significant arrhythmias recorded during ACM in these experienced pilots. PMID- 11396560 TI - The reduced oxygen breathing paradigm for hypoxia training: physiological, cognitive, and subjective effects. AB - The current training program for hypoxia familiarization requires a low-pressure chamber that places aviator trainees at risk for decompression sickness. A cost effective reduced oxygen-breathing (ROB) paradigm that decreases oxygen (O2) concentration leading to normobaric hypoxia was assessed as an alternative to the hypobaric chamber. PURPOSE: To help establish the validity of the ROB paradigm, this report documents cognitive performance, cardiopulmonary and subjective changes during ROB exposure. METHODS: Performance on a two-dimensional tracking task, as well as BP, heart rate, end-tidal carbon dioxide (ETCO2), O2 saturation, and subjective reports of hypoxia symptoms were observed in 12 U.S. Navy divers during exposure to normoxic air followed by one of four experimental gas mixtures per session. All participants received all gas conditions that differed in their relative concentrations of O2 and nitrogen (6.20/93.80, 7.00/93.00, 7.85/92.15, and 20.85/79.15% O2/N2). RESULTS: ROB caused increases in tracking task error (p < 0.0001). ROB also increased heart rate (p < 0.001) and systolic BP (p = 0.004), and decreased ETCO2 and O2 saturation (p < 0.0001). Finally, subjects responded to ROB-induced hypoxia with higher subjective symptom ratings (p < 0.0001). CONCLUSIONS: These data are consistent with those expected from hypoxic states and support the validity of the ROB paradigm for hypoxia training. Future validation studies comparing a ROB device with hypobaric chambers are needed. PMID- 11396561 TI - The basis for the development of a fuselage evacuation time for a ditched helicopter. AB - HYPOTHESIS: When a helicopter ditches or crashes in water, unless the buoyancy bags are inflated, it commonly sinks inverted. Thus, crew and passengers must make an underwater escape. It is postulated that later passengers in the escape sequence do not have the breath-holding ability to conduct a successful escape, particularly if the water is cold. This contributes to the 20-50% mortality rate in survivable accidents. METHODS: There were 132 immersed subject evaluations which were conducted in daylight and darkness to measure escape times from a helicopter underwater escape trainer, configured to the Super Puma, seated for 15 and 18 passengers. The subjects were highly experienced instructors or Navy clearance divers. RESULTS: The time from when each subject's head disappeared underwater until each subject surfaced and total fuselage evacuation time were measured and any problems hampering escape were noted. Breath-holding for the last subject out ranged from 28 to 92 s. An emergency breathing system was used by a minimum of four subjects each time and a maximum of 11 subjects in one condition. The buoyancy of the survival suit was the principal component that hampered escape. CONCLUSION: Breath-holding times were too long for the later subjects to escape without resorting to an EBS, in spite of the fact that they were highly trained. For regular crew and passengers flying over water, this would explain the high mortality, etc. Therefore, a new helicopter standard should be developed requiring fuselage design to accommodate total evacuation within 20 s from underwater. For current helicopters, where this cannot be achieved, passengers should be provided with some form of air supply, or, after ditching, the helicopter should be modified so that it will stay afloat on its side and retain an air space in the cabin. PMID- 11396562 TI - A proposed model for load carriage on sloped terrain. AB - BACKGROUND: The purpose of this study was to develop a predictive model for uphill and downhill load carriage. Relative to level walking, net energy costs increase with uphill movement and decrease moving downhill. To simulate load carriage over complex terrain, a model must estimate the cost of downhill movement. The net cost of downhill movement is expected to reach a minimum value, then increase as work is required to maintain stability. Thus, downhill costs cannot be simply extrapolated from a linear relationship for uphill work. METHOD: Oxygen uptake (VO2) was measured for 16 subjects during test sessions which consisted of walking at 1.34 m x s(-1) on a single grade (-12%, -10%, -8%, -4%, 2%, 0%, +4%, +4%, +8% and +12%) with a 0, 9.1- or 18.1-kg load. RESULTS: No significant gender differences were found, therefore data were pooled. The minimum VO2 values occurred at -8% grade. CONCLUSION: Our model assumes that the total energy requirement (WT) is the sum of the cost of level walking (W(L)) plus the cost of vertical displacement (Wv) for the total mass (body plus load). For uphill work, Wv was calculated by multiplying the cost of vertical displacement by an efficiency factor. For downhill work, the cost of vertical displacement was modified by an exponential function of the slope angle. Values for level and negative slope walking with no load were compared with estimated values derived from two published studies to partially validate the negative model. PMID- 11396563 TI - Peripheral effector mechanism hypothesis of postflight cardiovascular dysfunction. AB - Studies on the mechanisms of cardiovascular dysfunction after space-flight are important to illustrate the cardiovascular effect of microgravity and develop appropriate multi-system countermeasures for future long-duration spaceflights. Over the past 10 yr, we have systematically studied the adaptational changes in structure and function of both the heart and vessels, using the tail-suspension rat model to simulate microgravity effects. Our results indicate that simulated microgravity induced atrophic changes and reduced contractility of the heart muscle, and upward- and downward-regulation in structure, function, and innervation state of vessels in the brain and hind body of the rat. In addition, more recent advances in relevant ground-based and space-flight studies from different laboratories have also been reviewed. Based on these studies, it has been speculated that, in addition to hypovolemia, the microgravity-induced adaptational changes in the structure and function of the two main effectors of the cardiovascular system, i.e., the arterial smooth muscle and the cardiac muscle, might be among the most important mechanisms responsible for postflight cardiovascular dysfunction and orthostatic intolerance. In this paper we will review the available evidence with comments. PMID- 11396564 TI - Thermal properties of handwear at varying altitudes. AB - BACKGROUND: Total handwear insulation (I(T)) is dependent on the rate of heat transfer in air through the skin-handwear interface, handwear layers, and the surface boundary air layer. As altitude increases, the corresponding decrease in air pressure reduces convective heat loss. As convective heat losses decline, I(T), which is inversely related to the rate of heat loss, should increase. Increasing air velocity also reduces the insulation (Ia) provided by the boundary layer. METHODS: The military issue test handwear, Light-duty glove (LD), Trigger finger mitten (TF), and Arctic mitten (AM), were fitted over a biophysical hand model. Model surface temperatures were 25 degrees C, and air temperature was 10 degrees C. The handwear was tested at simulated altitudes of sea level (101 kPa), 2500 m (75 kPa) and 5000 m (54 kPa) in still air and at 5 m x s(-1). RESULTS: Overall, the effects of wind and altitude on I(T) were significant. Differences for I(T) between 0 and 5000 m were significant for LD and TF. Increases in I(T) greater than 10% are considered of sufficient magnitude to alter comfort sensation. CONCLUSIONS: Differences of that magnitude occurred most frequently between 0 and 5000 m. The present results are consistent with an increase in I(T) with increasing altitude. Changes in I(T) were greater in still air and for less insulated handwear where the contribution of Ia to I(T) was more important. PMID- 11396565 TI - On determining the relationship between blood alcohol concentration and smooth pursuit eye movements. AB - Residual impairment after alcohol consumption implies that the relationship between blood alcohol concentration (BAC) and deficits in performance of some task is not the same when becoming intoxicated as it is when returning to sobriety. A pilot study was performed to determine the feasibility of and the appropriate methodology for studies on residual impairment of smooth pursuit performance. Four subjects consumed alcohol for 2 h. Measurements of BAC and smooth pursuit eye movements were made every 30 min during drinking and for 4 h after drinking. Pursuits were elicited by having subjects track a sinusoidal target (0.40 Hz and 0.60 Hz) for 10 s. Impairment of smooth pursuit was quantified with frequency analysis scores. Frequency analysis scores declined as BAC increased. As BAC decreased, frequency analysis scores tended to increase toward pre-drink levels. The relationship between BAC and frequency analysis score was not significantly different on the ascending and descending limbs of the blood alcohol curve. However, the idea that residual impairment does not occur could not be conclusively demonstrated for several reasons. First, as BAC returned to pre-drink levels, frequency analysis scores were inconsistent for each subject perhaps because of boredom and fatigue. Second, the relationship between BAC and frequency analysis score varied between subjects. Finally, because the recording periods were short, the effect of alcohol on sustained attention could not be assessed. In this paper, potential artifacts in studies of residual impairment of pursuits are discussed and potential solutions to the methodological problems encountered in the pilot study are provided. PMID- 11396566 TI - Radiofrequency radiation overexposure. PMID- 11396567 TI - The evolution of bourgeois, parasitic, and cooperative reproductive behaviors in fishes. AB - Among vertebrate classes, fishes exhibit by far the greatest variability in competitive and cooperative behaviors in male reproduction. Scramble competition between reproductive males is one possibility. Another possibility occurs when resources, mates, or locations can be monopolized, in which case males may invest in primary access to fertilizations by adopting a "bourgeois" strategy, or they may employ alternative mating tactics to evade the reproductive monopoly of other males. Adaptations in morphology, physiology, and behavior to bourgeois and alternative phenotypes are highly divergent. Here I review the functional characteristics that differ between bourgeois and parasitic phenotypes, and discuss the variability of alternative reproductive tactics at the levels of plasticity, determination, and selection. Examples will illustrate the importance of ecology, and will suggest that variation in reproductive tactics is largely adaptive. Behavioral solutions to competition for mates and fertilizations often involve agonistic behavior and conflict, but also cooperation among competitors (e.g., when subordinate males pay a price to bourgeois males for gaining access to fertilizable eggs). Application of molecular genetic tools has helped to uncover intricate sexual and social relationships in various fish species, including species that display some of the most complex reproductive and social patterns known among the vertebrates. PMID- 11396568 TI - Genetic paternity analysis and breeding success in bluegill sunfish (Lepomis macrochirus). AB - Fish have some of the most complex mating systems known in the animal kingdom. With the advent of powerful genetic markers and an emerging mathematical framework to calculate parentage, it is now possible to analyze genetic relatedness and gene flow in these systems. An important example is the bluegill sunfish (Lepomis macrochirus) which consists of parental males that provide sole care for the young, cuckolder males that parasitize the parentals, and females that actively choose among males within dense breeding colonies. In this article genetic markers for bluegill are characterized and their utility in parentage studies is demonstrated by calculating the genetic relatedness of parental males to their broods for an entire natural breeding colony. A novel Monte Carlo simulation is developed to calculate the confidence in the relatedness estimates and these data are used to provide an estimate of the mean breeding success of parental and cuckolder males. Finally, the applications of genetic analyses to understanding mating systems, parental care, and life-history evolution in bluegill are discussed. PMID- 11396569 TI - Accuracy and precision of methods to estimate the number of parents contributing to a half-sib progeny array. AB - Molecular technologies have made feasible large-scale studies of genetic parentage in nature by permitting the genotypic examination of hundreds or thousands of progeny. One common goal of such studies is to estimate the true number of unshared parents who contributed to a large half-sib progeny array. Here we introduce computer programs designed to count the number of gametotypes contributed by unshared parents to each such progeny array, as well as assess the accuracy and precision of various estimators for the true number of unshared parents via computer simulation. These simulations indicate that under most biological conditions (1) a traditional approach (the multilocus MINIMUM METHOD) that merely counts the number of distinct haplotypes in offspring and divides by 2L, where L is the number of loci assayed, often vastly underestimates the true number of unshared parents who contributed to a half-sib progeny array; (2) a recently developed HAPLOTYPES estimator is a considerable improvement over the MINIMUM METHOD when parental numbers are high; and (3) the accuracy and precision of the HAPLOTYPES estimator increase as marker polymorphism and sample size increase, or as reproductive skew and the number of parents contributing to the progeny array decrease. Generally, HAPLOTYPES-based estimates of parental numbers in large half-sib cohorts should improve the characterization of organismal reproductive strategies and mating systems from genetic data. PMID- 11396570 TI - Kinship analysis of Pacific salmon: insights into mating, homing, and timing of reproduction. AB - Multilocus microsatellite genotypes were used to infer kinship and relatedness in two species of Pacific salmon from three populations in Washington State. Even in the absence of direct genetic data from parents, clustering of individuals according to allele sharing and reconstruction of parental genotypes allowed resolution of full- and half-sib relationships among 135 chinook salmon (Oncorhynchus tshawytscha) sampled as preemergent juveniles from 14 redds in the Dungeness River. Inferred reproductive behaviors included single-pair matings, polyandry in which females mated with two to three males at a single redd, polygyny in which males mated with two females at different redds, use of two redds by a single female, and use of one redd site by two females. Greater average relatedness (rxy) in the upper reach of the Dungeness River implied within-reach homing of returning adults. In steelhead trout (O. mykiss), the frequency of related pairs (dyads) of mature individuals that migrated up Snow Creek less than a week apart was greater than expected for randomly chosen dyads, as was the frequency of steelhead dyads that were spawned on the same day in the Forks Creek hatchery. These results imply a heritable basis for upstream migration date and maturation date in steelhead trout. PMID- 11396571 TI - A genetic evaluation of mating system and determinants of individual reproductive success in Atlantic salmon (Salmo salar L.). AB - The primary objective of this study was to use highly polymorphic microsatellite loci to estimate individual reproductive success in Atlantic salmon based on the number of surviving juveniles (young of the year) at the population level under natural conditions. We inferred reproductive strategies adopted by both sexes by applying a maximum likelihood method to determine parent-offspring genotype relationships. A high degree of variance in individual reproductive success for both males and females was revealed. The high number of mates used by both sexes is not concordant with previous behavioral studies proposing that females are mainly monogamous in this species. We found little evidence supporting the prediction from previous reports of a positive relationship between individual size and realized reproductive success for either males or females. For both sexes, however, there was a significant correlation between the number of mates and the number of offspring. These results indicate that this species' mating system is more flexible than previously thought and suggest that factors such as potential genetic benefits or environmental uncertainty may also be driving the evolution and the plasticity of mating systems in Atlantic salmon. PMID- 11396572 TI - Alternative mating strategies in Atlantic salmon and brown trout. AB - By screening variable number of tandem repeat (VNTR) loci, multiple paternity within clutches has been found in wild populations of southern European Atlantic salmon (Salmo salar) and brown trout (Salmo trutta). For Atlantic salmon, we determined the relative contribution of alternative male phenotypes to the next generation. Individual males that are morphologically juvenile yet sexually mature fertilized a large proportion of eggs, and they thereby contributed to an increase of genetic variability in wild populations via (1) balancing the sex ratio, (2) increasing outbreeding, and (3) enlarging the effective population size, in part a consequence of (1) and (2). In addition, these precocious males ensured that interspecific spawns involving Atlantic salmon females and brown trout males (a fairly common occurrence in southern Europe where the two species are sympatric) resulted mostly in Atlantic salmon progeny. For brown trout, preliminary genetic results indicated that multiple paternity, when present, was not due to alternative mating strategies by males, but rather to successive fertilizations by adult suitors. PMID- 11396573 TI - Mating systems and sexual selection in male-pregnant pipefishes and seahorses: insights from microsatellite-based studies of maternity. AB - In pipefishes and seahorses (family Syngnathidae), the males provide all postzygotic care of offspring by brooding embryos on their ventral surfaces. In some species, this phenomenon of male "pregnancy" results in a reversal of the usual direction of sexual selection, such that females compete more than males for access to mates, and secondary sexual characteristics evolve in females. Thus the syngnathids can provide critical tests of theories related to the evolution of sex differences and sexual selection. Microsatellite-based studies of the genetic mating systems of several species of pipefishes and seahorses have provided insights into important aspects of the natural history and evolution of these fishes. First, males of species with completely enclosed pouches have complete confidence of paternity, as might be predicted from parental investment theory for species in which males invest so heavily in offspring. Second, a wide range of genetic mating systems have been documented in nature, including genetic monogamy in a seahorse, polygynandry in two species of pipefish, and polyandry in a third pipefish species. The genetic mating systems appear to be causally related to the intensity of sexual selection, with secondary sex characters evolving most often in females of the more polyandrous species. Third, genetic studies of captive-breeding pipefish suggest that the sexual selection gradient (or Bateman gradient) may be a substantially better method for characterizing the mating system than previously available techniques. Finally, these genetic studies of syngnathid mating systems have led to some general insights into the occurrence of clustered mutations at microsatellite loci, the utility of linked loci in studies of parentage, and the use of parentage data for direct estimation of adult population size. PMID- 11396574 TI - Male pregnancy in seahorses and pipefishes (family Syngnathidae): rapid diversification of paternal brood pouch morphology inferred from a molecular phylogeny. AB - In contrast to the majority of vertebrate species, primary male parental care is common in fishes and encompasses a remarkable diversity of adaptations. Seahorses and pipefishes (Family Syngnathidae) exhibit some of the most specialized forms of paternal care in animals and so are ideally suited to the study of the evolution of male parental care. During mating, female syngnathids transfer eggs to specialized morphological structures that are located on either the abdomen or tail of the male. The male provides all postfertilization parental care and has morphological and physiological adaptations to osmoregulate, aerate, and even nourish the developing embryos. While all syngnathid species are adapted for paternal care, the brooding structure with which this is accomplished varies between species, from simple ventral gluing areas to much more complex structures such as the completely enclosed pouches of the seahorses. Our combined cytochrome b-, 12S rDNA-, and 16S rDNA-based molecular phylogeny of syngnathid fishes demonstrates that rapid diversification of male brooding structures has been associated with the major evolutionary radiation of the group, suggesting that development and diversification of structures involved in paternal care may have been key evolutionary innovations of the Syngnathidae. Molecular analyses also highlight geographical centers of biodiversity and suggest interoceanic migration of Syngnathus pipefishes from their center of origin in the Pacific. PMID- 11396575 TI - Genetic perspectives on the natural history of fish mating systems. AB - Molecular analyses of bird and mammal populations have shown that social mating systems must be distinguished from genetic mating systems. This distinction is important in fishes also, where the potential for extrapair spawning and intraspecific brood parasitism is especially great. We review studies on fishes that have used molecular markers to document biological parentage and genetic mating systems in nature, particularly in species with extended parental care of offspring. On average, nest-guarding adults parented about 70-95% of their custodial offspring, and approximately one-third of the nests were cuckolded to some extent. Furthermore, nearly 10% of the assayed nests contained offspring tended by foster fathers either because of nest takeovers or egg thievery. On average, fish that provide parental care on nests spawned with more mates than did fish with internal fertilization and pregnancy. Overall, genetic markers have both confirmed and quantified the incidence of several reproductive and other social behaviors of fishes, and have thereby enhanced our knowledge of piscine natural history. PMID- 11396576 TI - Insights into population ecology and sexual selection in snakes through the application of DNA-based genetic markers. AB - Hypervariable genetic markers have revolutionized studies of kinship, behavioral ecology, and population biology in vertebrate groups such as birds, but their use in snakes remains limited. To illustrate the value of such markers in snakes, we review studies that have used microsatellite DNA loci to analyze local population differentiation and parentage in snakes. Four ecologically distinct species of snakes all show evidence for differentiation at small spatial scales (2-15 km), but with substantial differences among species. This result highlights how genetic analysis can reveal hidden aspects of the natural history of difficult-to observe taxa, and it raises important questions about the ecological factors that may contribute to restricted gene flow. A 3-year study of genetic parentage in marked populations of the northern water snake showed that (1) participation in mating aggregations was a poor predictor of genetic-based measures of reproductive success; (2) multiple paternity was high, yet there was no detectable fitness advantage to multiple mating by females; and (3) the opportunity for selection was far higher in males than in females due to a larger variance in male reproductive success, and yet this resulted in no detectable selection on morphological variation in males. Thus genetic markers have provided accurate measures of individual reproductive success in this species, an important step toward resolving the adaptive significance of key features including multiple paternity and reversed sexual size dimorphism. Overall these studies illustrate how genetic analyses of snakes provide previously unobtainable information of long-standing interest to behavioral ecologists. PMID- 11396577 TI - Group-structured genetic models in analyses of the population and behavioral ecology of poikilothermic vertebrates. AB - Estimates of gene correlations among individuals within and among populations are frequently derived from statistical analyses of genetic data (e.g., F statistics). These measures can be important tools in molecular ecology and conservation, and offer important insights into population breeding structure. Using recently derived theory developed for group-structured populations, we show that fixation indices, when combined with basic population ecological and demographic data can be used to investigate population mating systems and to predict dispersal rates, trajectories and asymptotic levels of fixation indices, and effective population size. Four case studies of poikilothermic vertebrates are used to demonstrate the broad utility of evolutionary and ecological inferences afforded by group-structured models. PMID- 11396578 TI - Promiscuity in sand lizards (Lacerta agilis) and adder snakes (Vipera berus): causes and consequences. AB - We review postcopulatory phenomena in the Swedish sand lizard (Lacerta agilis) and adder (Vipera berus), and in particular, links between female promiscuity, determinants of paternity, and offspring viability. In both species, females mate multiply and exhibit a positive relationship between the number of partners and offspring viability. We conclude that this relationship is most likely the result of variable genetic compatibility between mates arising from postcopulatory phenomena, predominantly assortative fertilization with respect to parental genotypes. However, males who were more successful at mate acquisition were also more successful in situations of sperm competition, suggesting a possible link between male (diploid and haploid) genetic quality per se and probability of fertilization. Neither the number of partners nor the number of matings influenced the risk of infertility in sand lizards, suggesting that selection for reduced risk of infertility is not a sufficient explanation for maintaining female promiscuity in this population. Finally, we conclude that the relatively low genetic variability exhibited by our study populations may have facilitated detection of genetic benefits compared to more outbred ones. However, recent work derived from outbred populations in other taxa suggest a greater generality of the principles we discuss than previously may have been appreciated. PMID- 11396579 TI - The evolution of alternative reproductive strategies: fitness differential, heritability, and genetic correlation between the sexes. AB - Paternity analyses using molecular markers have become standard in studies of mating systems, parentage, and kinship. In systems where individuals exhibit alternative mating strategies, molecular analyses have been productively used to estimate the reproductive success of each behavioral type and hence the fitness consequences to each individual. Here we review the fitness results in a system of five alternative mating strategies present in one population of side-blotched lizards (Uta stansburiana). Males in this population adopt one of three behavioral strategies that differ in their degree of territoriality and mate guarding. In contrast, females adopt one of two strategies that differ in offspring quantity and quality. We use paternity analyses to estimate the fitness of each morph, the heritability of reproductive strategy, and the correlation in strategy between the sexes and discuss the implications of our findings for the evolution and maintenance of reproductive polymorphism in this and other systems. PMID- 11396580 TI - Turtle mating systems: behavior, sperm storage, and genetic paternity. AB - As evidenced by the articles in this volume, a recent increase in interest in the mating systems of poikilothermic vertebrates has focused primarily on fishes, a few amphibians, and squamate reptiles. Turtles by contrast have received relatively little attention, yet they display a wide variety of mating behaviors and life-history characteristics that make them excellent candidates for addressing several aspects of genetic parentage that should contribute to a broader understanding of animal reproductive strategies. Here we focus on genetic studies of the mating systems and reproductive patterns of turtles, specifically with respect to multiple paternity and long-term sperm storage. These phenomena highlight the importance of a temporally extended perspective on patterns of individual reproductive success. PMID- 11396581 TI - Molecular dating and biogeography of the early placental mammal radiation. AB - The timing and phylogenetic hierarchy of early placental mammal divergences was determined based on combined DNA sequence analysis of 18 gene segments (9779 bp) from 64 species. Using rooted and unrooted phylogenies derived from distinct theoretical approaches, strong support for the divergence of four principal clades of eutherian mammals was achieved. Minimum divergence dates of the earliest nodes in the placental mammal phylogeny were estimated with a quartet based maximum-likelihood method that accommodates rate variation among lineages using conservative fossil calibrations from nine different nodes in the eutherian tree. These minimum estimates resolve the earliest placental mammal divergence nodes at periods between 64 and 104 million years ago, in essentially every case predating the Cretaceous-Tertiary (K-T) boundary. The pattern and timing of these divergences allow a geographic interpretation of the primary branching events in eutherian history, likely originating in the southern supercontinent Gondwanaland coincident with its breakup into Africa and South America 95-105 million years ago. We propose an integrated genomic, paleontological, and biogeographic hypothesis to account for these earliest splits on the placental mammal family tree and address current discrepancies between fossil and molecular evidence. PMID- 11396582 TI - Rare variant of the intrasoleus musculocutaneous perforator: clinical considerations in raising a free peroneal osteocutaneous flap. AB - To improve the stability of the cutaneous portion of the vascularized free peroneal osteocutaneous flap (the so-called peroneal flap), various anatomic studies have been conducted, and a reliable notion of the course of cutaneous perforators has been provided; however, anatomic anomalies have occasionally been observed. The authors encountered a rare type of cutaneous perforator in the distal third of the lower leg. It was located under the inferior surface of the flexor hallucis longus muscle, after running in the soleus muscle parallel to the fibula, and then joined the posterior tibial artery. Since absolutely definite diagnostic methods have not been established for the course of the cutaneous perforator, surgery with the possibility of secondary anastomosis, always with a separate peroneal flap, should be a consideration. PMID- 11396583 TI - Coverage of a large soft-tissue defect of the chest with a free fillet forearm and hand flap. AB - The authors present a case in which a free circumferential fasciocutaneous flap from the forearm and hand, after radical tumor resection and forequarter amputation, was used successfully to cover the large soft-tissue defect on the chest wall. PMID- 11396584 TI - Burned ear reconstruction using a prefabricated free radial forearm flap. AB - Postauricular skin, postauricular fascia, temporoparietal fascia (pedicle or free), the free radial forearm flap (fascial or fasciocutaneous), and free lateral arm fascial flap, have been used for burned ear reconstruction reported in the literature. Patients who did not have normal tissue around the ear region, because of severe thermal injuries to the external ear, are not available for draping the costal cartilage framework; these patients require free flaps. The author reports a burned ear reconstruction, using a prefabricated free radial forearm fasciocutaneous flap, included an autogenous costal cartilage framework. In this case, the front and back of the cartilage framework were covered with the skin of the forearm flap. This is the main difference from other techniques in the literature. This flap is valuable for burned ear reconstruction, when local tissue and other free flaps are not available. PMID- 11396585 TI - Direct muscle neurotization after end-to-side neurorrhaphy. AB - Over the recent years, end-to-side neurorrhaphy has reemerged in the literature for reconstructive nerve surgery. Another technique, in which interest has remained continuous during the last century, is direct nerve implantation into muscle belly, so-called direct muscle neurotization (DMN). In this work, the authors present a new technique to recover muscle innervation through direct nerve implantation into muscle belly, by means of a nerve graft, and sutured with end-to-side neurorrhaphy. They carried out experiments on 20 Wistar rats divided into two groups. In Group 1 (10 rats), on the right side, the peroneal nerve was sutured to the tibial nerve with end-to-side neurorrhaphy. Subsequently, the terminal branches of the same nerve were implanted in the anterior tibial muscle (ATM). On the left side, the peroneal nerve was severed, and the ATM completely denervated. In Group 2 (10 rats), on the right side, the same technique was used as in first group. On the left side, the normal innervation of the ATM was maintained. After 4 months, all muscles and nerves were harvested and evaluations carried out on the morphologic aspect, weight, and histology of the ATM, as well as the histology of the nerves. The authors analyzed the results, which demonstrated good reinnervation of the muscles deprived of any nerve connection. PMID- 11396586 TI - IGF-I and end-to-side nerve repair: a dose-response study. AB - End-to-side nerve repair allows for target-muscle reinnervation, with simultaneous preservation of donor-nerve function. Local administration of insulin-like growth factor-I (IGF-I) has been shown to increase the rate of axon regeneration in crush-injured and freeze-injured rat sciatic nerve. The purpose of the current project was to determine the effects of IGF-I in a rat model of end-to-side nerve repair. The left musculocutaneous nerve of 18 adult male Sprague-Dawley rats was fully transected to induce biceps-muscle paralysis. The distal stump of the musculocutaneous nerve was then coapted by end-to-side neurorrhaphy through a perineurial window to the ipsilateral median nerve. All animals were randomly assigned to three groups: Group A received 100 microg/ml IGF-I; Group B received 50 microg/ml IGF-I; and control Group C received 10 mM acetic acid vehicle solution. Infusions were regulated by the Alzet model 2004 mini-osmotic pump, with an attached catheter directed at the coaptation site. Weekly postoperative behavioral evaluations revealed significantly increased functional return over control in both experimental groups as early as 3 weeks. After 28 days, histology evaluations revealed statistically significantly higher musculocutaneous nerve axon counts and myelin thickness/axon diameter ratios in both experimental groups vs. controls. The three groups were not significantly different in motor endplate counts of the biceps muscle. Groups A and B were not significantly different in all parameters tested. This study suggests that local infusion of IGF-I may expedite the functional recovery of a paralyzed muscle, by increasing the rate of axon regeneration through an end-to-side nerve graft. PMID- 11396587 TI - Fibrin sealant: a novel method of fixation for an implantable ultrasonic microDoppler probe. AB - While free tissue transfer affords the reconstructive microvascular surgeon the ability to provide coverage for complex wounds, the postoperative monitoring of these flaps continues to evolve. The most recent advance has been the development of an implantable microDoppler probe to provide an early warning signal for vascular obstruction. The current system relies on the use of a silicone cuff to secure a 1-mm probe to the outflow vein. The release force to remove the probe from the cuff is reported to be 1/10 of a pound (45 g). A disadvantage of this system is the need for a circumferential, relatively inelastic device around the vein. Should the cuff be too tightly secured to the vein, the potential for outflow obstruction exists. Moreover, if the probe is not well-approximated to the vein, no signal is produced. Finally, the fact that a foreign body remains in the wound after completion of the monitoring period remains a concern. The authors have investigated a new method to adhere the probe, using a commercially available fibrin sealant. The use of this biocompatible substance has the potential to obviate the need for the current method of fixation, and the associated concerns. PMID- 11396588 TI - Side-to-side arterial anastomosis model in the rat internal and external carotid arteries. AB - Microvascular anastomosis has become a general and essential technique in the field of vascular reconstructive surgery. Side-to-side microvascular anastomosis is rarely performed by microsurgeons. Laboratory training models are essential for the development and refinement of microsurgical techniques. A new experimental model of a side-to-side "arterial" anastomosis in the rat internal and external carotid arteries is presented, in which 100 percent patency was accomplished immediately, and was maintained 7 days after anastomosis. This model can provide an experimental and training tool for side-to-side anastomosis of small arteries. PMID- 11396589 TI - Development of a mouse model for heterotopic limb and composite-tissue transplantation. AB - The mouse remains the most suitable model to study the complexities of the immune system and transplant rejection. The purpose of this study was to describe a new mouse model for heterotopic limb and composite tissue transplantation. Eighteen procedures were performed, including 10 heterotopic lower hind limb, four vascularized skin, and four vascularized muscle transplantations. Three transplants were allogeneic, and the rest were syngeneic. All successful syngeneic transplants were harvested at 11 days postoperatively, except for one skin and one limb transplant that were followed for over 30 days. The allogeneic transplants showed signs of rejection between 7 to 11 days postoperatively. Results of mixed lymphocyte culture (p < 0.05) and histology evaluations from the allogeneic recipients were consistent with acute rejection as the cause of allograft loss. The mortality rate was 16.7 percent, and the overall success rate was 72.2 percent. Details of the operative procedure are described, and important technical factors are discussed. PMID- 11396590 TI - Different methods and results in the treatment of obstetrical brachial plexus palsy. PMID- 11396591 TI - A novel robust index to assess beat-to-beat variability in heart rate time-series analysis. AB - A new index is proposed to estimate the variance of the differentiated heart rate (RR) time series from its truncated histogram. The index is more robust to artifacts than the standard deviation of the differentiated RR time series (rMSDD) and, unlike the pNN50, does not saturate for very high or very low heart rate variability. PMID- 11396592 TI - Accurate identification of periodic oscillations buried in white or colored noise using fast orthogonal search. AB - We use a previously introduced fast orthogonal search algorithm to detect sinusoidal frequency components buried in either white or colored noise. We show that the method outperforms the correlogram, modified covariance autoregressive (MODCOVAR) and multiple-signal classification (MUSIC) methods. Fast orthogonal search method achieves accurate detection of sinusoids even with signal-to-noise ratios as low as -10 dB, and is superior at detecting sinusoids buried in 1/f noise. Since the utilized method accurately detects sinusoids even under colored noise, it can be used to extract a 1/f noise process observed in physiological signals such as heart rate and renal blood pressure and flow data. PMID- 11396593 TI - Electric field stimulation of cardiac myocytes during postnatal development. AB - Studies on cardiac cell response to electric field stimulation are important for understanding basic phenomena underlying cardiac defibrillation. In this work, we used a model of a prolate spheroidal cell in a uniform external field (Klee and Plonsey, 1976) to predict the threshold electric field (ET) for stimulation of isolated ventricular myocytes of rats at different ages. The model assumes that ET is primarily determined by cell shape and dimensions, which markedly change during postnatal development. Neonatal cells showed very high ET, which progressively decreased with maturation (experimental mean values were 29, 21, 13, and 5.9 and 6.3 V/cm for 3-6, 13-16, 20-21, 28-35, and 120-180 day-old rats, respectively, P < 0.001; theoretical values were 24, 18, 11, 9, and 6 V/cm, respectively). Estimated maximum membrane depolarization at threshold (deltaVT approximately equals 35 mV, under our experimental conditions) was reasonably constant during development, except for cells from 1-mo-old animals, in which deltaVT was lower than at other ages. We conclude that the model reasonably correlates ET with cell geometry and size in most cases. Our results might be relevant for the development of efficient procedures for defibrillation of pediatric patients. PMID- 11396594 TI - A novel approach for precise simulation of the EMG signal detected by surface electrodes. AB - We propose a new electromyogram generation and detection model. The volume conductor is described as a nonhomogeneous (layered) and anisotropic medium constituted by muscle, fat and skin tissues. The surface potential detected in space domain is obtained from the application of a two-dimensional spatial filter to the input current density source. The effects of electrode configuration, electrode size and inclination of the fibers with respect to the detection system are included in the transfer function of the filter. Computation of the signal in space domain is performed by applying the Radon transform; this permits to draw considerations about spectral dips and clear misunderstandings in previous theoretical derivations. The effects of generation and extinction of the action potentials at the fiber end plate and at the tendons are included by modeling the source current, without any approximation of its shape, as a function of space and time and by using again the Radon transform. The approach, based on the separation of the temporal and spatial properties of the muscle fiber action potential and of the volume conductor, includes the capacitive tissue properties. PMID- 11396595 TI - Finite-element analysis of aortic valve-sparing: influence of graft shape and stiffness. AB - Aortic valve incompetence due to aortic root dilation may be surgically corrected by resuspension of the native valve within a vascular graft. This study was designed to examine the effect of graft shape and material properties on aortic valve function, using a three-dimensional finite-element model of the human aortic valve and root. First, the normal root elements in the model were replaced with graft elements, in either a cylindrical or a "pseudosinus" shape. Next, the elements were assigned the material properties of either polyethylene terephthalate, expanded polytetrafluoroethylene, or polyurethane. Diastolic pressures were applied, and stresses, strains, and coaptation were recorded for the valve, root, and graft. Regarding shape, the cylindrical graft models increased the valve stresses by up to 173%, whereas the root-shaped graft model increased valve stresses by up to 40% as compared to normal. Regarding material properties, the polyurethane models demonstrated valve stress, strain, and coaptation values closest to normal, for either root shape. Graft shape had a greater effect on the simulated valve function than did the material property of the graft. Optimizing the shape and material design of the graft may result in improved longevity of the spared valve if a normal environment is restored. PMID- 11396596 TI - Localization of the site of origin of cardiac activation by means of a heart model-based electrocardiographic imaging approach. AB - We have developed a new approach to solve the inverse problem of electrocardiography in terms of heart model parameters. The inverse solution of the electrocardiogram (ECG) inverse problem is defined, in the present study, as the parameters of the heart model, which are closely related to the physiological and pathophysiological status of the heart, and is estimated by using an optimization system of heart model parameters, instead of solving the matrix equation relating the body surface ECGs and equivalent cardiac sources. An artificial neural network based preliminary diagnosis system has been developed to limit the searching space of the optimization algorithm and to initialize the model parameters in the computer heart model. The optimal heart model parameters were obtained by minimizing the objective functions, as functions of the observed and model-generated body surface ECGs. We have tested the feasibility of the newly developed technique in localizing the site of origin of cardiac activation using a pace mapping protocol. The present computer simulation results show that, the present approach for localization of the site of origin of ventricular activation achieved an averaged localization error of about 3 mm [for 5-muV Gaussian white noise (GWN)] and 4 mm (for 10-muV GWN), with standard deviation of the localization errors of being about 1.5 mm. The present simulation study suggests that this newly developed approach provides a robust inverse solution, circumventing the difficulties of the ECG inverse problem, and may become an important alternative to other ECG inverse solutions. PMID- 11396597 TI - Estimating the posterior probability of LTP failure by sequential Bayesian analysis of an imperfect Bernoulli trial model. AB - A tetanically stimulated (TS) neuron is said to have failed to fire if its voltage-clamped excitatory postsynaptic current (EPSC) measurement is devoid of a long-term potentiation (LTP) response. This paper provides a method for evaluating the posterior probability of "failure" for TS neurons. A sequential Bayes algorithm is employed on an imperfect Bernoulli trial model to refine the posterior with each EPSC data record processed. The method is applied to both real and simulated LTP data and is shown to be consistent with the theoretical Beta-distributed posterior and the reported in vitro voltage-clamped EPSC failure rates. PMID- 11396598 TI - Improved coil design for functional magnetic stimulation of expiratory muscles. AB - Our studies have demonstrated effective stimulation of the expiratory muscles in patients with spinal cord injury (SCI) using functional magnetic stimulation (FMS). The observed contraction of the expiratory muscles and functional improvement of the pulmonary functions make functional magnetic stimulation an appropriate tool for expiratory muscle training. To fully capitalize on the benefits of FMS for expiratory muscle training, this study aimed to optimize the magnetic coils (MCs). The primary goal of this study was to investigate how two parameters of the MC size and winding structure, would affect expiratory muscle training. By varying these parameters, our approach was to conceptualize and evaluate the induced electric field and nerve activation function distributions of six coils, round 9.2, 13.7, and 20 cm, and spiral 9.2-, 13.7-, and 20-cm coils in the computer modeling phase. Round 9.2 cm, spiral 13.7 cm, and spiral 20-cm coils were also evaluated in experimental studies for induced electrical field and in clinical studies of expiratory muscles. Both the computer models and experimental measurements indicated that the spiral 20-cm coil can not only stimulate more expiratory spinal nerves but can also stimulate them more evenly. In addition, coils with larger diameters had better penetration than those with smaller diameters. The clinical results showed that the spiral 20-cm coil produced higher expiratory pressure, flow, and volume in five able-bodied subjects, and it was the coil of choice among the subjects when asked their preferences. In our attempt to optimize MC design for FMS of expiratory muscle training, we followed the designing guidelines set out in our previous study and arrived at a more effective tool. PMID- 11396599 TI - A low-cost fiber-optic instrument to colorimetrically detect patients with Barrett's esophagus for early detection of esophageal adenocarcinoma. AB - In Barrett's esophagus, the precursor to esophageal adenocarcinoma, the squamocolumnar junction (SCJ) between the normal esophagus and the stomach like mucosa is proximally displaced. Currently it can be detected only by an expensive upper GI endoscopic procedure. We have developed a minimally invasive and easy to operate colorimetric instrument for the low-cost detection of Barrett's esophagus. The instrument is based on a flexible, narrow diameter, fiber-optic probe that performs a colorimetric scan of the esophageal lumen. The instrument was clinically evaluated in 50 subjects. The instrument could identify both symmetric and asymmetric SCJ's. The SCJ locations determined by the colorimetric instrument correlated strongly (R2 = 0.89) with those determined by endoscopy. The instrument identified the SCJ locations accurately (Mean of difference +/- SEM: 0.97 +/- 1.72 cm) and reproducibly (Mean of absolute difference +/- SEM: 1.33 +/- 1.40 cm). The instrument has a 90% sensitivity of identifying patients with Barrett's esophagus, based on the clinical algorithm that if the SCJ is located at a distance less than 37 cm from the teeth, then the subject has Barrett's esophagus, otherwise the subject does not have Barrett's esophagus. In conclusion, the colorimetric instrument has the potential of being a cost effective way of determining patients likely to have Barrett's esophagus in the population. PMID- 11396600 TI - Computer-assisted quantification of axo-somatic boutons at the cell membrane of motoneurons. AB - This paper presents a system for computer-assisted quantification of axo-somatic boutons at motoneuron cell-surface membranes. Different immunohistochemical stains can be used to prepare tissue of the spinal cord. Based on micrographs displaying single neurons, a finite element balloon model has been applied to determine the exact location of the cell membrane. A synaptic profile is extracted next to the cell membrane and normalized with reference to the intracellular brightness. Furthermore, a manually selected reference cell is used to normalize settings of the microscope as well as variations in histochemical processing for each stain. Thereafter, staining, homogeneity, and allocation of boutons are determined automatically from the synaptic profiles. The system is evaluated by applying the coefficient of variation (Cv) to repeated measurements of a quantity. Based on 1856 motoneuronal images acquired from four animals with three stains, 93% of the images are analyzed correctly. The others were rejected, based on process protocols. Using only rabbit anti-synaptophysin as primary antibody, the correctness increases above 96%. Cv values are below 3%, 5%, and 6% for all measures with respect to stochastic optimization, cell positioning, and a large range of microscope settings, respectively. A sample size of about 100 is required to validate a significant reduction of staining in motoneurons below a hemi-section (Wilcoxon rank-sum test, alpha = 0.05, beta = 0.9). Our system yields statistically robust results from light micrographs. In future, it is hoped that this system will substitute for the expensive and time-consuming analysis of spinal cord injury at the ultra-structural level, such as by manual interpretation of nonoverlapping electron micrographs. PMID- 11396601 TI - Automatic identification of human helminth eggs on microscopic fecal specimens using digital image processing and an artificial neural network. AB - In order to automate routine fecal examination for parasitic diseases, we propose in this study a computer processing algorithm using digital image processing techniques and an artificial neural network (ANN) classifier. The morphometric characteristics of eggs of human parasites in fecal specimens were extracted from microscopic images through digital image processing. An ANN then identified the parasite species based on those characteristics. We selected four morphometric features based on three morphological characteristics representing shape, shell smoothness, and size. A total of 82 microscopic images containing seven common human helminth eggs were used. The first stage (ANN-1) of the proposed ANN classification system isolated eggs from confusing artifacts. The second stage (ANN-2) classified eggs by species. The performance of ANN was evaluated by the tenfold cross-validation method to obviate the dependency on the selection of training samples. Cross-validation results showed 86.1% average correct classification ratio for ANN-1 and 90.3% for ANN-2 with small variances of 46.0 and 39.0, respectively. The algorithm developed will be an essential part of a completely automated fecal examination system. PMID- 11396602 TI - Line patterns in the mosaic electrical properties of human skin--a cross correlation study. AB - A vehicle with 16 electrodes for the two-dimensional electrical admittance mapping of human skin is presented. Measurements on 20 test subjects have been carried out and analyzed for the possible detection of line patterns in the electrical properties of the skin, claimed to coincide with the so-called acupuncture meridian lines of ancient Chinese medicine. No such lines were found. PMID- 11396603 TI - An ultralight biotelemetry backpack for recording EMG signals in moths. AB - A two-channel FM biopotential recording system fabricated on a foldable, lightweight, polyimide substrate is presented. Each channel consists of a biopotential amplifier followed by a Colpitts oscillator with operating frequency tunable in the 88-108 MHz commercial FM band. The overall system measures 10 mm X 10 mm X 3 mm, weighs 0.74 g, uses two 1.5-V batteries, dissipates about 2 mW, and has a transmission range of 2 m. Using this system, electromyogram signals have been recorded from the dorsal ventral muscle and the dorsal longitudinal muscle of a giant sphinx moth (manduca sexta). PMID- 11396604 TI - Estimated generalized least squares electromagnetic source analysis based on a parametric noise covariance model. AB - Estimated generalized least squares (EGLS) electromagnetic source analysis is used to downweight noisy and correlated data. Standard EGLS requires many trials to accurately estimate the noise covariances and, thus, the source parameters. Alternatively, the noise covariances can be modeled parametrically. Only the parameters of the model describing the noise covariances need to be estimated and, therefore, less trials are required. This method is referred to as parametric egls (PEGLS). In this paper, PEGLS is developed and its performance is tested in a simulation study and in a pseudoempirical study. PMID- 11396606 TI - Glycine neurotransmitter transporters: an update. AB - Glycine accomplishes several functions as a transmitter in the central nervous system (CNS). As an inhibitory neurotransmitter, it participates in the processing of motor and sensory information that permits movement, vision, and audition. This action of glycine is mediated by the strychnine-sensitive glycine receptor, whose activation produces inhibitory post-synaptic potentials. In some areas of the CNS, glycine seems to be co-released with GABA, the main inhibitory amino acid neurotransmitter. In addition, glycine modulates excitatory neurotransmission by potentiating the action of glutamate at N-methyl-D-aspartate (NMDA) receptors. It is believed that the termination of the different synaptic actions of glycine is produced by rapid re-uptake through two sodium-and-chloride coupled transporters, GLYT1 and GLYT2, located in the plasma membrane of glial cells or pre-synaptic terminals, respectively. Glycine transporters may become major targets for therapeutic of pathological alterations in synaptic function. This article reviews recent progress on the study of the molecular heterogeneity, localization, function, structure, regulation and pharmacology of the glycine transporter proteins. PMID- 11396605 TI - Multiplicity and regulation of genes encoding peptide transporters in Saccharomyces cerevisiae. AB - The model eukaryote Saccharomyces cerevisiae has two distinct peptide transport mechanisms, one for di-/tripeptides (the PTR system) and another for tetra /pentapeptides (the OPT system). The PTR system consists of three genes, PTR1, PTR2 and PTR3. The transporter (Ptr2p), encoded by the gene PTR2, is a 12 transmembrane domain (TMD) integral membrane protein that translocates di /tripeptides. Homologues to Ptr2p have been identified in virtually all organisms examined to date and comprise the PTR family of transport proteins. In S. cerevisiae, the expression of PTR2 is highly regulated at the cellular level by complex interactions of many genes, including PTR1, PTR3, CUP9 and SSY1. Oligopeptides, consisting of four to five amino acids, are transported by the 12 14 TMD integral membrane protein Opt1p. Unlike Ptr2p, distribution of this protein appears limited to fungi and plants, and there appears to be three paralogues in S. cerevisiae. This transporter has an affinity for enkephalin, an endogenous mammalian pentapeptide, as well as for glutathione. Although it is known that OPT1 is normally expressed only during sporulation, to date little is known about the genes and proteins involved in the regulation of OPT1 expression. PMID- 11396607 TI - The amino acid transport system b(o,+) and cystinuria. AB - Amino acid transport in mammalian plasma membranes is mediated by a multiplicity of amino acid transport systems. Some of them (systems L, y+ L, x(c)- and b(o,+)) are the result of the activity of heteromeric amino acid transporters (HAT) (i.e. transport activity is elicited by the coexpression of a heavy and a light subunit). The two heavy subunits known today (HSHAT: rBAT and 4F2hc) were identified in 1992, and light subunits (LSHAT: LAT-1, LAT-2, asc-1, y+ LAT-1, y+ LAT-2, xCT and b(o,+)AT) have been cloned in the last 2 years. Defects in two genes of this family (SLC3A1, encoding rBAT and SLC7A9, encoding b(o,+)AT) are responsible for cystinuria, an inherited aminoaciduria of cystine and dibasic amino acids. This finding and functional studies of rBAT and b(o,+)AT suggested that these two proteins encompassed the high-affinity renal reabsorption system of cystine. In contrast to this view, immunofluorescence studies showed that rBAT is most abundant in the proximal straight tubule, and b(o,+)AT is most abundant in the proximal convoluted tubule of the nephron. The need for a new light subunit for rBAT and a heavy subunit for b(o,+)AT is discussed. PMID- 11396608 TI - The role of system A for neutral amino acid transport in the regulation of cell volume. AB - System A is a secondary active, sodium dependent transport system for neutral amino acids. Strictly coupled with Na,K-ATPase, its activity determines the size of the intracellular amino acid pool, through a complex network of metabolic reaction and exchange fluxes. Many hormones and drugs affect system A activity in specific cell models or tissues. In all the cell models tested thus far the activity of the system is stimulated by amino acid starvation, cell cycle progression, and the incubation under hypertonic conditions. These three conditions produce marked alterations of cell volume. The stimulation of system A activity plays an important role in cell volume restoration, through an expansion of the intracellular amino acid pool. Under normal conditions, system A substrates represent a major fraction of cell compatible osmolytes, organic compounds that exert a protein stabilizing effect. It is, therefore, likely that the activation of system A represents a portion of a more complex response triggered by exposure to stresses of various nature. Since system A transporters have been recently cloned, the molecular bases of these regulatory mechanisms will probably be elucidated in a short time. PMID- 11396609 TI - Molecular mechanisms in proximal tubular and small intestinal phosphate reabsorption (plenary lecture). AB - Renal and small intestinal (re-)absorption contribute to overall phosphate(Pi) homeostasis. In both epithelia, apical sodium (Na+)/Pi-cotransport across the luminal (brush border) membrane is rate limiting and the target for physiological/pathophysiological alterations. Three different Na/Pi cotransporters have been identified: (i) type I cotransporter(s)--present in the proximal tubule--also show anion channel function and may play a role in secretion of organic anions; in the brain, it may serve vesicular glutamate uptake functions; (ii) type II cotransporter(s) seem to serve rather specific epithelial functions; in the renal proximal tubule (type Ila) and in the small intestine (type IIb), isoform determines Na+-dependent transcellular Pi movements; (iii) type III cotransporters are expressed in many different cells/tissues where they could serve housekeeping functions. In the small intestine, alterations in Pi-absorption and, thus, apical expression of IIb protein are mostly in response to longer term (days) situations (altered Pi intake, levels of 1.25 (OH2) vitamin D3, growth, etc), whereas in renal proximal tubule, in addition, hormonal effects (e.g. Parathyroid Hormone, PTH) acutely control (minutes/hours) the expression of the IIa cotransporter. The type II Na/Pi-cotransporters operate (as functional monomers) in a 3 Na+:1 Pi stoichiometry, including transfer of negatively charged (-1) empty carriers and electroneutral transfers of partially loaded carriers (1 Na+, slippage) and of the fully loaded carriers (3 Na+, 1 Pi). By a chimera (IIa/IIb) approach, and by site-directed mutagenesis (including cysteine-scanning), specific sequences have been identified contributing to either apical expression, PTH-induced membrane retrieval, Na+-interaction or specific pH-dependence of the IIa and IIIb cotransporters. For the COOH-terminal tail of the IIa Na/Pi-cotransporter, several interacting PDZ-domain proteins have been identified which may contribute to either its apical expression (NaPi-Cap1) or to its subapical/lysosomal traffic (NaPi-Cap2). PMID- 11396610 TI - Syntaxin 1A up-regulates GABA transporter expression by subcellular redistribution. AB - Neurotransmitter transporters are regulated through a variety of signal transduction mechanisms which appear to operate in order to maintain appropriate levels of transmitter in the synaptic cleft. One such mechanism is the trafficking of the transporter in association with synaptic vesicle release machinery. This report examines the specifics of trafficking regulation of the rat brain GABA transporter GAT1 by syntaxin 1A, a plasma membrane component of the SNARE complex which is involved in vesicle membrane fusion. In hippocampal neurons, botulinum neurotoxin 1C, which specifically cleaves syntaxin 1A, down regulates plasma membrane GAT1 levels as assessed by surface biotinylation, suggesting that syntaxin 1A is a positive regulator of GAT1 surface expression. The up-regulation correlates with a decrease in intracellular GAT1 levels and is complete within several minutes. These data suggest that syntaxin 1A mediates the redistribution of GAT1 on a time scale important for the rapid regulation of extracellular GABA levels. Expression of different syntaxin 1A constructs in Xenopus oocytes suggests that several portions of the syntaxin 1A molecule are required for the trafficking of GAT1. These data suggest that the trafficking of GAT1 will be subject to regulatory control by the many molecules known to interact with various domains of syntaxin 1A. PMID- 11396611 TI - Serotonin and norepinephrine transporters: possible relationship between oligomeric structure and channel modes of conduction. AB - In the past several years there has been significant progress made on the biophysics of neurotransmitter transporters, leading to the proposal of new models of substrate and ion permeation across membranes. Questions arising from these studies are as follows: How are substrate uptake and substrate-induced current related? Where and how does substrate-ion coupling occur? What is the functional significance of the coupled and uncoupled currents? Because of a long standing interest and collaboration, and because of their importance for normal function and disease, the authors have focused on the properties of human norepinephrine and serotonin transporters, using other clones and mutations as specific needs arise. It has been know for decades that hNETs (human norepinephrine transporters) clear NE+ (norepinephrine) following its release in peripheral sympathetic and central noradrenergic synapses. Neuronal activity influences NE+ uptake, so one is also interested in the acute regulation of hNET. To study these problems, hNET-expressing cells have been developed that are suitable for patch clamp, radioligand uptake, biochemistry, and transiently expressed clones for structure-function analysis, and new protocols have been designed combining patch-clamp, microamperometry, Ca2+ imaging, and native catecholamine transporter preparations to study transporters in whole cells and isolated patches. Using these methods, Na-dependent, NE+-induced hNET currents that are blocked by cocaine and antidepressants, channel modes of NE+ conduction, voltage-dependent uptake coupled to NE+-induced ion channel activity, PKC (phosphokinase C) regulation of NE+ uptake, and transporter modulation by [Ca2+]i have all been discovered. There is also provocative new data on other transporters in this family, such as Li/Na mole fraction experiments in the Drosophila serotonin transporters and sided enkephalin block in proline transporters. These studies have led one to postulate the existence of a narrow pore within transporters through which the substrate (NE+ or serotonin, 5HT+) and other ions (principally Na+) pass. It is hypothesized that the pore resides in an oligomeric structure and that separate gene products of hNET or hSERT (human serotonin transporters) come together to form a channel. PMID- 11396612 TI - The ENT family of eukaryote nucleoside and nucleobase transporters: recent advances in the investigation of structure/function relationships and the identification of novel isoforms. AB - The first examples of the equilibrative nucleoside transporter (ENT) family were characterized in human tissues at the molecular level only 4 years ago. Since that time, the identification of homologous proteins by functional cloning and genome analysis has revealed that the family is widely distributed in eukaryotes. Family members are predicted to possess 11 transmembrane helices (TMs), and recent investigations on the mammalian ENTs have implicated the TM 3-6 region in solute recognition. Whilst the name of the family reflects the properties of its prototypical member hENT1, an equilibrative transporter of nucleosides, some family members can also transport nucleobases and some are proton-dependent, concentrative transporters. In addition to their role in nucleoside salvage, ENTs are targets for coronary vasodilator drugs and act as routes for uptake of cytotoxic drugs in humans and protozoa. This paper summarizes current knowledge of the family and reports on the identification of a novel mammalian ENT isoform, designated ENT3, from mouse and human tissues. PMID- 11396613 TI - Recent molecular advances in studies of the concentrative Na+-dependent nucleoside transporter (CNT) family: identification and characterization of novel human and mouse proteins (hCNT3 and mCNT3) broadly selective for purine and pyrimidine nucleosides (system cib). AB - The human concentrative (Na+-linked) plasma membrane transport proteins hCNT1 and hCNT2, found primarily in specialized epithelia, are selective for pyrimidine nucleosides (system cit) and purine nucleosides (system cif), respectively. Both have orthologs in other mammalian species and belong to a gene family (CNT) that also includes members in lower vertebrates, insects, nematodes, pathogenic yeast and bacteria. The CNT transporter family also includes a newly identified human and mouse CNT3 transporter isoform. This paper reviews the studies of CNT transport proteins that led to the identification of hCNT3 and mCNT3, and gives an overview of the structural and functional properties of these latest CNT family members. hCNT3 and mCNT3 have primary structures that place them in a CNT subfamily separate from CNT1/2, transport a wide range of physiological pyrimidine and purine nucleosides and antineoplastic and antiviral nucleoside drugs (system cib), and exhibit a Na+:uridine coupling ratio of at least 2:1 (cf 1:1 for hCNT1/2). Cells and tissues containing hCNT3 transcripts include mammary gland, differentiated HL-60 cells, pancreas, bone marrow, trachea, liver, prostrate and regions of intestine, brain and heart. In HL-60 cells, hCNT3 is transcriptionally regulated by phorbol myristate (PMA). The hCNT3 gene, which contains an upstream PMA response element, mapped to 9q22.2 (cf chromosome 15 for hCNT1 and hCNT2). PMID- 11396614 TI - Functional production of mammalian concentrative nucleoside transporters in Saccharomyces cerevisiae. AB - The transport of nucleosides and nucleobases in the yeast Saccharomyces cerevisiae is reviewed and the use of this organism to study recombinant mammalian concentrative nucleoside transport (CNT) proteins is described. A selection strategy based on the ability of an expressed nucleoside transporter cDNA to mediate thymidine uptake by yeast under a selective condition that depletes endogenous thymidylate was used to assess the transport capacity of heterologous transporter proteins. The pyrimidine-nucleoside selective concentrative transporters from human (hCNT1) and rat (rCNT1) complemented the imposed thymidylate depletion in S. cerevisiae, as did N-terminally truncated versions of hCNT1 and rCNT1 lacking up to 31 amino acids. Transporter-mediated rescue of S. cerevisiae by both nucleoside transporters was inhibited by cytidine, uridine and adenosine, but not by guanosine or inosine. This work represents the development of a new model system for the functional production of recombinant nucleoside transporters of the CNT family of membrane proteins. PMID- 11396615 TI - Complex regulation of nucleoside transporter expression in epithelial and immune system cells. AB - Nucleoside transporters have a variety of functions in the cell, such as the provision of substrates for nucleic acid synthesis and the modulation of purine receptors by determining agonist availability. They also transport a wide range of nucleoside-derived antiviral and anticancer drugs. Most mammalian cells co express several nucleoside transporter isoforms at the plasma membrane, which are differentially regulated. This paper reviews studies on nucleoside transporter regulation, which has been extensively characterized in the laboratory in several model systems: the hepatocyte, an epithelial cell type, and immune system cells, in particular B cells, which are non-polarized and highly specialized. The hepatocyte co-expresses at least two Na+-dependent nucleoside transporters, CNT1 and CNT2, which are up-regulated during cell proliferation but may undergo selective loss in certain experimental models of hepatocarcinomas. This feature is consistent with evidence that CNT expression also depends on the differentiation status of the hepatocyte. Moreover, substrate availability also modulates CNT expression in epithelial cells, as reported for hepatocytes and jejunum epithelia from rats fed nucleotide-deprived diets. In human B cell lines, CNT and ENT transporters are co-expressed but differentially regulated after B cell activation triggered by cytokines or phorbol esters, as described for murine bone marrow macrophages induced either to activate or to proliferate. The complex regulation of the expression and activity of nucleoside transporters hints at their relevance in cell physiology. PMID- 11396616 TI - Vitamin C transport systems of mammalian cells. AB - Vitamin C is essential for many enzymatic reactions and also acts as a free radical scavenger. Specific non-overlapping transport proteins mediate the transport of the oxidized form of vitamin C, dehydroascorbic acid, and the reduced form, L-ascorbic acid, across biological membranes. Dehydroascorbic acid uptake is via the facilitated-diffusion glucose transporters, GLUT 1, 3 and 4, but under physiological conditions these transporters are unlikely to play a major role in the uptake of vitamin C due to the high concentrations of glucose that will effectively block influx. L-ascorbic acid enters cells via Na+ dependent systems, and two isoforms of these transporters (SVCT1 and SVCT2) have recently been cloned from humans and rats. Transport by both isoforms is stereospecific, with a pH optimum of approximately 7.5 and a Na+:ascorbic acid stoichiometry of 2:1. SVCT2 may exhibit a higher affinity for ascorbic acid than SVCT1 but with a lower maximum velocity. SVCT1 and SVCT2 are predicted to have 12 transmembrane domains, but they share no structural homology with other Na+ co transporters. Potential sites for phosphorylation by protein kinase C exist on the cytoplasmic surface of both proteins, with an additional protein kinase A site in SVCT1. The two isoforms also differ in their tissue distribution: SVCT1 is present in epithelial tissues, whereas SVCT2 is present in most tissues with the exception of lung and skeletal muscle. PMID- 11396617 TI - Multidrug transporters in prokaryotic and eukaryotic cells: physiological functions and transport mechanisms. AB - Multidrug transporters mediate the extrusion of structurally unrelated drugs from prokaryotic and eukaryotic cells. As a result of this efflux activity, the cytoplasmic drug concentration in the cell is lowered to subtoxic levels and, hence, cells become multidrug resistant. The activity of multidrug transporters interferes with the drug-based control of tumours and infectious pathogenic microorganisms. There is an urgent need to understand the structure-function relationships in multidrug transporters that underlie their drug specificity and transport mechanism. Knowledge about the architecture of drug and modulator binding sites and the link between energy-generating and drug translocating functions of multidrug transporters may allow one to rationally design new drugs that can poison or circumvent the activity of these transport proteins. Furthermore, if one is to inhibit multidrug transporters in human cells, one should know more about their physiological substrates and functions. This review will summarize important new insights into the role that multidrug transporters in general, and P-glycoprotein and its bacterial homologue LmrA in particular, play in the physiology of the cell. In addition, the molecular basis of drug transport by these proteins will be discussed. PMID- 11396618 TI - Thoracoscopy as a nonpharmacotherapeutic research modification for limiting postoperative chest pain. AB - Diminished tissue injury and shortened clinical recovery are benefits of using an endoscopic approach for patients needing operative procedure. In the course of developing an experimental model requiring procurement of topographically precise lung biopsy specimens, we sought to apply thoracoscopy as a research alternative to thoracotomy. In addition, we investigated the influence of thoracoscopy on postprocedure recovery practices using rabbits divided into four treatment groups. Rabbit groups 1 and 2 underwent thoracoscopy and lung biopsy while maintained by one-lung anesthesia. Additionally, group 2 had ketoprofen and bupivacaine HCl analgesics injected for treatment during postprocedure recovery. These two groups were compared to control rabbits in groups 3 and 4, which underwent inhalant anesthesia without thoracoscopy. Control group 3 also received the injection analgesic combination. During recovery, rabbit behavior was systematically assessed for evidence of pain. No behavior considered indicative of pain needing intervention was observed regardless of treatment group. Limited changes in plasma corticosterone, catecholamines, and prostaglandin E2 levels measured during recovery were difficult to associate with any treatment. Unexpectedly, significantly different mean corticosterone and catecholamines levels were detected in rabbits given the injection analgesic combination in the absence of thoracoscopic procedure, as compared to other treatment groups. The results highlight the importance of awareness that analgesic drug administration has the potential to alter homeostasis and affect interpretation of some study findings by its own guise. Correlation of the mean pain study results with plasma biochemical data supports preferential use of thoracoscopy as a refinement for limiting postprocedural pain in research models. PMID- 11396619 TI - Chronic osteomyelitis in a new rabbit model. AB - Dogs, rats, and rabbits are the most suitable species to induce chronic osteomyelitis and to study different methods of treatment. In rabbits, the incidence of and mortality from Staphylococcus aureus-induced osteomyelitis of the tibia depends on the method of prelesions and the amount and virulence of species-specific bacteria used. In this study two different lesions were combined simultaneously in the medullary canal of the femurs by aspiration of bone marrow, leaving the insertion needle in situ. A sclerosing agent was then inoculated followed by 300,000 bacteria of a rabbit-derived S. aureus strain to initiate infection. With this method, the incidence of chronic progressive osteomyelitis of the femur was increased to 100%. A relatively low mortality was observed, probably due to a lower number of inoculated bacteria as compared to other rabbit models described. The incidence of acute to chronic osteomyelitis was diagnosed by local signs, x-rays, microbiological recovery, and gross pathology of the femur. Initial fever, weight loss, abscess formation in soft tissues, and pain on palpation characterize the clinical features in the course of development of this chronic disease. PMID- 11396620 TI - Piaget and his role in problem based learning. PMID- 11396621 TI - The long journey to cardiac transplantation. PMID- 11396622 TI - Surgical management of dyslipidemia: clinical and experimental evidence. AB - Coronary artery disease (CAD) is still a major cause of mortality in developed countries, and dyslipidemia is one of its major causes. In an attempt to reduce both mortality and morbidity from CAD, several dietary, pharmacological, and surgical approaches have been used to reduce plasma cholesterol levels. In this brief review, we summarize the evidence for cholesterol-lowering effects and safety of partial ileal bypass (PIB) procedure in both human and animal studies. The results of the Program on the Surgical Control of the Hyperlipidemias (POSCH), which involved a total of 838 subjects with myocardial infarction, are promising. A 5-year follow-up of this study revealed significant reductions of up to 27% in total cholesterol (TC) and up to 42% in low-density lipoprotein (LDL) cholesterol levels along with an increase of up to 8% in high-density lipoprotein (HDL) cholesterol levels as compared to controls. These changes were associated with other benefits such as increased HDL/TC and HDL/LDL ratios, and a significant decrease in apolipoprotein (apo) B100 and increase in apo AI levels. Similar results were also demonstrated by other studies. PIB surgery is one of the most effective methods for reduction of plasma cholesterol levels, particularly in patients with heterozygous familial hypercholesterolemia. This procedure is also applicable to treatment of sitosterolemia, a rare genetic disorder in which the absorption of plant sterols is abnormally high. Although no major complications of this method have been reported, more extensive studies are required to evaluate its long-term effects on renal and hepatic function. Similarly, long-term impact of this procedure on progression/regression of atherosclerotic lesions must be documented. Finally, indications for this procedure should be carefully considered, particularly in view of availability of other treatments of dyslipidemia. PMID- 11396623 TI - Porcine orthotopic liver autotransplantation: facilitated technique. AB - The technique of a liver autograft in the pig has three advantages: (1) It provides an excellent training model of liver transplantation, (2) it provides an experimental model for cancer research, and (3) it is more economical than liver allotransplant. We describe a facilitated technique of liver autograft, which can be employed to develop experimental models without the use of a biopump. Mean blood arterial pressure, heart rate, pH, and lactates were tested during the liver grafting and at the end of the procedure in pigs that underwent autografting of the liver and compared with pigs that underwent an orthotopic liver allotransplantation. The cell damage was assessed in the same two groups of animals by monitoring aspartate aminotransferase (AST) and alanine aminotransferase (ALT) blood levels and with the MEGX test, 15 min after the beginning of reperfusion. The surgical procedure may be divided into three parts: hepatectomy, side-to-side portocaval shunt with passive caval-jugular shunt, and reimplantation. This procedure could have a clinical indication for otherwise unresectable liver tumors. PMID- 11396624 TI - Effect of lazaroid U-74389G on pulmonary ischemia-reperfusion injury in dogs. AB - Lipid peroxidation induced by oxygen free radicals is a contributing factor in ischemia-reperfusion injury. Lazaroid U-74389G (LAZ-G) is a group of new synthetic 21-aminosteroids and inhibits irondependent lipid peroxidation. We investigated the effects of LAZ-G on pulmonary ischemia-reperfusion injury in dogs. Twenty dogs were divided into three groups. In the LAZ-G group (n = 6), LAZ G was administered 15 min before ischemia. In the St group (n = 5), methylprednisolone was injected 15 min before ischemia and 15 min before reperfusion. In the control group (n = 9), the vehicle of Lazaroid was injected 15 min before ischemia. Warm ischemia was induced for 3 h by clamping the pulmonary artery and veins. Arterial oxygen saturation (SaO2), cardiac output (CO), left pulmonary vascular resistance (L-PVR), and blood levels of interleukin 1beta mRNA were measured. The lung specimen was harvested for histologic study and polymorphonuclear neutrophils (PMNs) counting. SaO2 levels at 30 min and 2 h after reperfusion were significantly higher in the LAZ-G group than in the control group. After 30 min of reperfusion, CO was significantly better in the LAZ-G group than in the St and control groups, and the L-PVR level was significantly lower in the LAZ-G group than in the control group. Survival rates were significantly better in the LAZ-G group than in the control group. Histological damages and PMNs infiltration were more severe in the control group than in the LAZ-G group. Interleukin-1beta mRNA levels were lower in the LAZ-G group than in the control group. Lazaroid U-74389G appears to generate a protective effect against ischemia-reperfusion injury of the lung. PMID- 11396625 TI - Prevention of pericardial adhesions with N-O carboxymethylchitosan in the rabbit model. AB - The presence of mediastinal adhesions significantly increases the morbidity and mortality of reoperative cardiac surgical procedures. Previous investigations have reported on the therapeutic utility of topical hydrogels in reducing the formation of postsurgical adhesions. The goal of the present study is to evaluate the ability of N-O carboxymethylchitosan (a glycosaminoglycan hydrogel derivative) to reduce the formation ofpostsurgical pericardial adhesions in a large-animal model. Sixteen adult New Zealand white rabbits were randomly assigned to one of two treatment groups. Group 1 subjects (n = 8) had N-O carboxymethylchitosan directly applied to the heart and retrosternal surfaces after sternotomy was performed, while subjects in group 2 (n = 8) had saline applied to these areas. After a period of 14 days the animals were sacrificed under anesthesia, and independent observers, blinded to treatment, graded the formation of pericardial adhesions. The severity of adhesion formation was significantly less in the group treated with N-O carboxymethylchitosan (p < .01). This study demonstrates that N-O carboxymethylchitosan markedly decreases the formation of poststernotomy adhesions in a large-animal model without untoward cardiac side effects. This hydrogel derivative may prove to be of great therapeutic value when used prophylactically in the setting of cardiac surgery. PMID- 11396626 TI - Neutrophil depletion and chemokine response after liver ischemia and reperfusion. AB - Neutrophils play a major role in the hepatic microvasculature following liver ischemia and reperfusion (I/R). Leukocyte cytokine chemoattractants (chemokines) are produced by neutrophils and cause neutrophil activation in I/R injury. We examined the role of neutrophils in the production of chemokines in the liver and lung inflammatory response following liver I/R. C57BL/6 mice were subjected to partial liver ischemia for 90 min. Four groups of animals were included: sham group, sham group with neutrophil depletion, ischemic control group, and ischemic control with neutrophil depletion. We evaluated at 3 h liver injury measurements, serum macrophage inflammatory protein-2 (MIP-2) and macrophage inflammatory protein-1 alpha (MIP-1alpha) chemokines, liver and lung tissue myeloperoxidase (MPO), and liver and lung histology. Statistical analysis included analysis of variance (ANOVA), and Student-Newman-Keuls and Kruskal-Wallis multiple comparison Z-value tests. Ischemic controls showed a significant increase in liver enzyme levels along with statistically significant higher liver and lung MPO activity values than the rest of the other groups (p < .05). MIP-2 values were higher in the ischemic control group when compared to the ischemic neutrophil depleted group. MIP-1alpha levels showed opposite results, being significantly lower (p < .05) in the ischemic control as compared to the neutrophil-depleted group. Improved liver and lung histopathological features were observed in the ischemic neutrophil depleted group when compared to the ischemic control group. Our study confirmed the key role of neutrophils in liver I/R injury and appeared to suggest some relationship between neutrophils and the production of certain chemokines, such as MIP-1alpha, which had an inverse relationship in the absence of neutrophils. Further studies will confirm the validity of these preliminary observations. PMID- 11396627 TI - Antigen retrieval immunohistochemistry and molecular morphology in the year 2001. AB - The impact of the antigen retrieval (AR) technique upon diagnostic immunohistochemistry (IHC) and upon research has been demonstrated by numerous of articles and more than a dozen major reviews. The specific aim of this survey of the field is to examine potential new approaches to retrieval of nucleic acid and protein from archival paraffin-embedded tissue for molecular biology-based diagnostic procedures that form the basis of the emerging field of molecular morphology. Any new approach must incorporate the principles of standardization and improved reproducibility. The ultimate goal will be to understand the mechanisms of fixation (by formalin) and "unfixation" (by AR). In the interim, the diligent application of practical procedures that have been shown to be tried and true is the least that we must demand from ourselves and our laboratories. PMID- 11396628 TI - Follicular dendritic cells in reactive and neoplastic lymphoid tissues: a reevaluation of staining patterns of CD21, CD23, and CD35 antibodies in paraffin sections after wet heat-induced epitope retrieval. AB - Structural alterations in the meshwork of follicular dendritic cells (FDCs) are frequently found in malignant lymphomas. Formaldehyde fixation and paraffin embedding, however, have long prevented consistent detection of FDCs. Wet heat induced epitope retrieval in Dako Target Retrieval Solution (TRS) (pH 6.0) enabled the reliable detection of FDCs through CD21, CD23, and CD35 antigens in routinely processed tissues from 11 reactive and 69 neoplastic lymphoproliferations, thus allowing the distribution of the FDCs to be reevaluated. Germinal center FDCs in lymphoid hyperplasias and expanded FDC meshworks in the 8 mantle cell lymphomas, 7 low-grade MALT lymphomas, and 6 low grade follicular lymphomas were intensely stained with all these markers. In 6 cases of B cell chronic lymphocytic leukemia, tumor cells were CD23+. In four cases of nodular lymphocyte predominance Hodgkin's disease (HD), expanded FDC meshwork's sharply delineating negative tumor cells and their rosetting T cell, were revealed mainly with the CD21 and CD35 antibodies. Follicular dendritic cells were also demonstrated in 11 cases of grade I nodular sclerosing HD, including follicular HD. Striking dendritic cell clusters were revealed with all 3 antibodies in 9 angioimmunoblastic T cell lymphomas. Sparse or no FDC meshworks were detected in the 4 cases of grade II nodular sclerosing HD, 5 follicular lymphomas with high-grade transformation, and 5 T cell-rich B cell lymphomas. CD35 immunostaining showed the most consistent labeling in the four FDC sarcomas studied in the current article. Reproducible demonstration of FDCs in routinely processed paraffin sections with CD21, CD23, and CD35 antibodies, as presented here, provides invaluable pieces of information in the diagnosis of lymphoproliferative disorders. PMID- 11396629 TI - CD99 immunoreactivity in ependymoma. AB - The expression of CD99 in normal ependymal cells and ependymoma has been reported. However, only limited numbers of tumors have been studied, and the pattern of CD99 expression has not been described. The authors' purpose was to investigate CD99 immunoreactivity in ependymoma and its use for differential diagnosis. Twenty-five ependymomas were immunostained with antibody directed at CD99. The result of immunostaining of ependymomas was compared with 63 nonependymal tumors that histologically resemble ependymal neoplasms. The nonependymal tumors included 19 astrocytic tumors, 6 oligodendroglial tumors, 8 choroid plexus neoplasms, 2 central neurocytomas, 5 medulloblastomas, 10 primitive neuroectodermal tumors (PNET), and 13 pituitary adenomas. All ependymomas showed strong expression of CD99 in membranous pattern with intracytoplasmic or intercellular dots (ICDs). The expression pattern of CD99 was not correlated with histologic type or grade of ependymomas. Among 63 nonependymal tumors, 11 (17.5%) showed incomplete membrane staining for CD99; diffuse in 4 PNETs and focal in 5 choroid plexus neoplasms (3 papillomas and 2 carcinomas) and one each of pituitary adenoma and oligodendroglioma. The ICD was not found in nonependymal tumors except a case of choroid plexus papilloma. However, membrane staining or ICD for CD99 was not distinctive in nonependymal tumors. In conclusion, the characteristic pattern of anti-CD99 antibody, i.e., diffuse strong membranous immunostaining with ICDs, is useful in distinguishing ependymomas from the central nervous system tumors that histologically mimic ependymoma. PMID- 11396630 TI - MMP-3 mRNA and MMP-3 and MMP-1 proteins in bladder cancer: a comparison with clinicopathologic features and survival. AB - Matrix metalloproteinases (MMPs) are proteolytic enzymes important at several points during multistep neoplastic progression. Although MMP-1 and MMP-3 have been implicated in the progression of various human cancers, their expression in bladder cancer has not been addressed. Immunohistochemistry (Strept-ABC-HRP method) and in situ hybridization were performed to detect MMP-1 protein, MMP-3 protein, and MMP-3 mRNA, respectively, in 59 transitional cell bladder carcinomas. To assess the role of these MMPs in bladder cancer, their expression was evaluated in relation to known clinicopathologic parameters and patients' disease-free and overall survival. Immunoreactivity for MMP-1 and MMP-3 proteins was observed in the cytoplasm of cancer cells in 30.5% and 24% of samples, respectively. Transcripts for MMP-3 mRNA were localized in stromal cells in 71.2% of cases and in cancer cells in 49% of cases. MMP-1 immunoreactivity demonstrated a statistically significant association with deeply invasive and grade III tumors versus superficial and lower grade tumors. MMP-3 protein immunoreactivity and MMP 3 mRNA immunolocalization did not associate with the parameters studied. However, MMP-3 mRNA localization in stromal cells demonstrated a borderline association with poor patients' disease-free and overall survival. In conclusion, the authors' results demonstrate a differential expression between MMP-1 and MMP-3 in bladder cancer; MMP-1 appears to participate in invasiveness and possibly in loss of differentiation in urothelial carcinomas in contrast to MMP-3. PMID- 11396631 TI - CD44V6 in gastric carcinoma: a marker of tumor progression. AB - CD44 is a group of cell surface molecules involved in cell-cell and cell-matrix interactions. CD44 spliced variants (CD44V) have been found to enhance the metastatic potential of rat tumors. Tumors from the breast, colon, and thyroid express many alternatively spliced products; nonneoplastic tissues do not. Some authors suggest that CD44V5 and V6 may play a role in gastric carcinoma. The aim of the current study was to investigate the role of CD44V6 as a prognostic marker and predictor of metastatic potential in gastric carcinomas. One hundred fifty five cases of gastric adenocarcinomas were studied: 36 cases of early (EGC), 19 cases of intermediate (MGC), and 100 cases of advanced gastric adenocarcinomas (AGC). A monoclonal antibody against CD44V6 (R&D) was used. CD44V6 expression was positively correlated with advanced stage (P = 0.05). Strong positivity was only detected in those cases of AGC with metastases. Patients with CD44V6 positive tumors revealed a lower 3- and 5-year survival rate (P = 0.0002). Immunohistochemical detection of CD44V6 could now be used as an indicator of tumor progression in biopsies of patients with gastric carcinoma. PMID- 11396632 TI - Expression of survivin, YB-1, and KI-67 in sporadic adenomas and dysplasia associated lesions or masses in ulcerative colitis. AB - Sporadic adenomas are said to exhibit an orderly growth pattern with a reversal of proliferative and apoptotic cell distribution as compared with normal colonic crypts. Dysplastic polyps of patients with ulcerative colitis (UC) may represent dysplasia-associated lesions or masses (DALM) with a high associated cancer risk, or, alternatively, may represent sporadic adenomas. Histologic criteria to differentiate between sporadic adenomas and DALM have not focused on the balance between cell renewal and cell loss. The expression of the novel anti-apoptosis gene product, survivin, and the proliferation markers, Ki-67 and Y-box binding protein (YB-1), were investigated by immunohistochemical localization in sporadic adenomas and DALM lesions of patients with UC. In adenomas, KI-67 was expressed preponderantly at the luminal aspect of the polyp, whereas its expression was diffuse in DALM. Survivin was detected diffusely in both adenomas and DALM. YB-1 showed positive staining in the deep aspect of adenomatous glands but only to a minor degree at the surface, whereas both deep and diffuse expression patterns of YB-1 were seen in DALM. The authors conclude that DALM and sporadic adenomas exhibit different patterns of cellular proliferation and that molecular markers of cell proliferation, Ki-67 and YB-1, may be useful to distinguish sporadic adenomas from DALM. However, the similar expression of survivin suggests that the underlying mechanisms that regulate apoptotic cell death are uniform in these lesions. PMID- 11396633 TI - Concordant p53 and mdm-2 protein expression in vulvar squamous cell carcinoma and adjacent lichen sclerosus. AB - To determine if carcinogenic events in vulvar skin precede the onset of morphologic atypia, the authors investigated for derangements in DNA content, cell proliferation, and cell death in vulvar carcinomas and surrounding skin in 140 samples of tumor and surrounding skin collected from 35 consecutive vulvectomy specimen for squamous cell carcinoma (SCC) or vulvar intraepithelial neoplasia (VIN) 3. Vulvar non-cancer excisions were used as controls. Investigations consisted of histologic classification and measurement of 9 variables--epidermal thickness (acanthosis and rete ridge length), immunolabeling index (LI) for 3 proteins (p53 protein, Ki-67, and mdm-2), pattern of p53 expression (dispersed vs. compact), DNA content index, and presence of aneuploidy by image analysis and apoptotic rate by Apotag labeling. Significant positive correlations were found for all nine variables studied versus increasing histologic severity in two proposed histologic stepwise models of vulvar carcinogenesis (lichen sclerosus (LS) and VIN 3 undifferentiated associated SCC groups). High p53 LI (>25) and the compact pattern of p53 expression (suspected oncoprotein) significantly correlated with LS and its associated vulvar samples compared with samples not associated with LS (P < or = 0.001). Furthermore, p53 LI, mdm-2 LI, and pattern of p53 expression were concordant between patient matched samples of LS and SCC. In addition, mdm-2 LI significantly correlated with dispersed pattern p53 LI suggesting a response to wild-type p53 protein accumulation. These findings support the hypothesis that neoplastic transformation occurs in sequential steps and compromises proteins involved in the cell cycle control. Concordance of p53 and mdm-2 protein expression in LS and adjacent SCC provides evidence that LS can act as a precursor lesion in the absence of morphologic atypia. Overexpression of mdm-2 with stabilization and inactivation of p53 protein may provide an alternate pathway for vulvar carcinogenesis. PMID- 11396634 TI - Immunohistochemical distinction of endometrial stromal sarcoma and cellular leiomyoma. AB - Distinguishing low grade endometrial stromal sarcoma (ESS) from benign smooth muscle proliferations like cellular leiomyoma (CL) can be problematic; because of differing treatments and prognosis, this distinction is important. The authors tested the hypothesis that low grade ESS could be distinguished from CL by immunohistochemistry using a panel of antibodies that have not previously been used in this setting. Antibodies to calponin, smooth muscle myosin heavy chain (SMM-HC), the Wilms tumor gene product (WT-1), and CD10 were applied to 14 cases of ESS (10 low grade, 4 high grade) and 9 CL. Among low grade ESS, 3 of 10, 3 of 10, 9 of 10, and 10 of 10 were positive for expression of calponin, SMM-HC, WT-1, and CD10, respectively. Of CL, all 9 were positive for calponin, SMM-HC, and WT 1, whereas 3 of 9 marked with antibodies to CD10. Overall, SMM-HC and calponin were expressed strongly in CL but weakly expressed in ESS; the converse was true for CD10. Expression of WT-1 and the reticulin-staining pattern do not discriminate between these two tumors. Antibodies to SMM-HC, CD10, and calponin can reliably distinguish ESS from CL. PMID- 11396635 TI - TGF-beta isoform and receptor expression in giant cell tumor and giant cell lesions of bone. AB - The authors examined the distribution of tumor growth factor-beta (TGF-beta) isoforms and receptors in 35 giant cell tumor (GCT) of bone in comparison with a group of benign giant cell-containing lesions of bone, including 5 aneurysmal bone cysts, 2 cases of brown tumor of hyperparathyroidism, 3 nonossifying fibromas, and 7 cases of giant cell reparative granuloma. The results of immunohistochemical analysis of GCT showed a complete absence of TGF-beta1 expression in both mononuclear tumor cells and giant cells. Only reactive bone present within the tumor showed an intense immunoreactivity. Transforming growth factor-beta2 and TGF-beta3 were detected in the majority of cases (97.1% and 82.8%, respectively), whereas TGF-beta receptor type I (TGF-beta RI) and type II (TGF-beta RII) were diffusely expressed in all cases. Reverse transcription polymerase chain reaction (RT-PCR) analysis performed on 10 GCTs with specific oligonucleotide primers demonstrated the presence of mRNA transcripts for TGF beta1, 2, 3, and for TGF-beta RI and RII. Quantitative measurements of TGF-beta1 in conditioned media from primary cultures of GCT showed undetectable or very low amounts of the cytokine (0-23 pg/mL). The results of immunohistochemical analysis showed that all giant cell-containing lesions of bone were at least focally positive for the 3 isoform of TGF-beta, with positivity present both in osteoclast-like giant cells and mononuclear cells, and diffusely positive for TGF beta RI and RII. Reverse transcription-polymerase chain reaction analysis conducted on samples from 3 nonossifying fibromas and 1 giant cell reparative granuloma confirmed the expression of the corresponding mRNA. In conclusion, according to the current data, GCT of bone can be distinguished from other giant cell-containing lesions of bone on the basis of the absence of TGF-beta1 expression at the protein level, which appears to be the result of posttranslational regulation processes. PMID- 11396636 TI - Formalin-fixed and heat-retrieved tissue antigens: a comparison of their immunoreactivity in experimental antibody diluents. AB - Heat-induced antigen retrieval procedures have become an effective and therefore important part of immunohistochemistry. Although used with increasing frequency, it is not known by what mechanism or to what extent these procedures restore the molecular integrity of the antigen. In an attempt to explore these questions, the immune reactions of 14 tissue antigens with their corresponding monoclonal antibodies were compared both in the absence and after the application of two widely used heat-induced antigen retrieval procedures. The tissue antigens were selected either because of the increased or lack of response to heat retrieval. The nature of their immune reactions was appraised through the variable effects that some diluent solutes were previously shown to have on monoclonal antibodies at different pH. It was found that on heat retrieval, four antigens reacting with monoclonal antibodies of subclass IgG2a or IgG1 reversed their preference for an alkaline pH to that of an acid pH. Eight of nine antigens reacting with monoclonal antibodies of subclass IgG1 and one antigen reacting with an antibody of class IgM remained most reactive at pH 6.0. The retrieved epitope of a B-cell antigen reacting with an IgG3 subclass antibody retained its preference for the alkaline pH. These patterns of response by the antigens to heat-induced retrieval are compared with the previously reported modifications in the immune reactions of monoclonal antibodies when sodium chloride and sodium phosphates were included in the antibody diluent buffer. Use of antibody diluents containing these solutes in combination with heat-induced antigen retrieval resulted in the loss of most of the sensitivity gained through this technique. The possible common mechanisms underlying the causes for these results are discussed. PMID- 11396637 TI - Expression of CD44 and its isoforms in the fetal neuroblasts. AB - CD44 is a polymorphic transmembrane glycoprotein that exists as multiple isoforms resulting from alternative splicing and posttranslational modifications. Enhanced expression of CD44 has been correlated to the tumorigenicity and metastatic behavior in different malignant tumors. In contrast, human neuroblastomas exhibit an inverse correlation between CD44 expression and tumor malignancy. To determine whether there is a CD44 silencing in sympatho-adrenal precursors as a possible explanation for the down-regulation of CD44 in neuroblastomas, the expression of standard CD44H and v6, v7, v7v8, or v10 isoforms was analyzed by immunohistochemistry in human adrenal glands of 14- to 20-week-old gestational age fetuses. All of the fetal neuroblasts localized in the adrenal gland parenchyma and migrating from the sympathetic nerve chain into the fetal adrenal cortex strongly expressed CD44H but none of the CD44 isoforms could be detected in these cells. In contrast, a strong expression of CD44v7 and v6 was detected in the fetal adrenal cells. These results indicate that, as for many other cell types, the CD44H expressed by fetal neuroblasts may contribute to controlling their migration into the adrenal medulla and that the down-regulation of CD44H in neuroblastoma cells should be explained by mechanisms other than the fetal suppression of CD44H expression in their normal counterparts. PMID- 11396638 TI - Ki67 reactivity in cell membrane and cytoplasm of tonsil crypt epithelial cells. PMID- 11396639 TI - CD79: a review. AB - CD79 is composed of CD79a and CD79b components expressed almost exclusively on B cells and B-cell neoplasms. CD79a and CD79b expression precedes immunoglobulin (Ig) heavy-chain gene rearrangement and CD20 expression during B-cell ontogeny and disappears later than CD20 in the late (plasma cell) stage of B-cell differentiation. Therefore, antibodies to CD79a and CD79b are useful in the differential diagnosis of B-cell neoplasms from T-cell neoplasms or myeloid neoplasms, or L and H lymphocyte predominance Hodgkin's lymphoma from classic Hodgkin's lymphoma. In addition, CD79a and CD79b antibodies are useful markers in the diagnosis of precursor B-acute lymphoblastic leukemia (pre-B-ALL) because many of these tumors are negative for other B-cell markers, such as CD20 and CD45RA. Furthermore, for B-cell neoplasms, wherein CD20 expression is aberrantly lost, such as in diffuse large B-cell lymphoma, or for B-cell neoplasms after CD20-antibody therapy, CD79a may be used as a first-line B-cell marker for the diagnosis. In this review, the authors discuss the molecular biology of CD79 and the frequency and usefulness of CD79 expression in these neoplasms. PMID- 11396641 TI - Recent advancements in the treatment of epithelial ovarian cancer. AB - Important advances in the management of advanced epithelial ovarian cancer have been made in recent years, with much of the knowledge emanating from clinical trials conducted by the Gynecological Oncology Group (GOG). This monograph reviews the trials that have defined current clinical practice and summarizes some innovative techniques and promising new drugs for the future PMID- 11396640 TI - Distribution of oxygenated blood flow at three different routes of extracorporeal membrane oxygenation in exteriorized fetal lambs. AB - OBJECTIVE: The purpose of this study was to determine which extracorporeal membrane oxygenation (ECMO) route best approximates the normal physiologic pattern of oxygenated blood distribution in fetal lambs submerged in warm saline solution. STUDY DESIGN: Six fetal lambs ranging from 113 to 129 days of gestation were delivered by cesarean section and oxygenated with ECMO in a warm saline solution bath. We compared the distribution of oxygenated blood flow during intact placental circulation with 3 routes of ECMO, from the right atrium to the carotid artery (V-A), from the right atrium to umbilical vein (V-UV), and from the right atrium with umbilical artery to umbilical vein [(V + UA)-UV], in exteriorized fetal lambs. Distribution of oxygenated blood flow was determined by nonradioactive colored microspheres. RESULTS: The ECMO blood flow rate through the catheters during V-A, V-UV or (V + UA)-UV ECMO ranged at 150 to 300 ml/min. Among the 3 different routes of ECMO, the oxygenated blood distribution in (V + UA)-UV showed the best correlation with placental-fetal blood flow, particularly in the brain and heart. CONCLUSION: This study indicated that (V + UA)-UV ECMO most closely approximates normal intact placental-fetal blood flow distribution in fetal lambs. PMID- 11396642 TI - Gravid uterus in an incisional hernia. AB - A case of gravid uterus in an incisional hernia is reported. The pregnancy was further complicated by intrauterine growth restriction, oligohydramnios and pregnancy induced hypertension. The management of the case is discussed. PMID- 11396643 TI - Intracranial venous thrombosis associated with severe antithrombin-III deficiency in pregnancy. AB - We report a patient with intracranial venous thrombosis in the third trimester of pregnancy associated with severe antithrombin-III deficiency. The evaluation of protein C, protein S and antithrombin-III levels in patients with thrombotic events during pregnancy may reveal the specific cause of the thrombotic event and thereby influence patient management PMID- 11396644 TI - Splenectomy combined with cesarean section in a patient with severe immunological thrombocytopenic purpura refractory to medical therapy. AB - Splenectomy as a treatment for severe immunological thrombocytopenic purpura is rarely performed in the third trimester of pregnancy. We report a 28-year-old patient who presented at the 34th gestational week with severe immunological thrombocytopenic purpura resistant to both corticosteroid and intravenous immunoglobulin therapies. Splenectomy combined with delivery by cesarean section provided remission of the disease. PMID- 11396645 TI - Legal and ethical considerations of assisted reproductive technology and surrogate motherhood in AOFOG countries. AB - With increasing use of assisted reproductive technologies (ART) and its related issues, the Ethics Committee of the Asia-Oceania Federation of Obstetrics and Gynecology (AOFOG), after discussion at its council meeting, took up the mission to understand the current legal and ethical aspects of this special subject in Asia-Oceania area before making proper recommendations. This Committee drafted a questionnaire to all 22 country member societies in 1995, and again conducted the same survey in 2000 to track changes in each country/region's laws and regulations. The results show that the current attitude and status of country member societies vary in a wide range and do not have formal legal codes in majority regulating the ethical aspects of ART. The use of ART and surrogacy is obviously affected by local traditions, popularity in the use of such techniques and cultural/religious concerns. The changes in 5 years are not major. Three countries do not as yet practice such techniques. However, liberalization of surrogate motherhood has become a trend. Five countries, compared with two only in 1995, are now available for surrogate motherhood, though regulated by strict criteria. Based on the data collected and issues discussed, a detailed guideline may not be generally applicable as each region has its own rules, reflecting differences in the management of ART, especially in the surrogate motherhood. This Committee therefore recommends that ART must be performed by a board certified gynecologist in a certification laboratory and surrogate motherhood must be used only for medical reasons and not be commercialized. The patients of this special group must be treated individually, based on this recommendation and the regulations of their own country/region. PMID- 11396647 TI - Gonadotropin-releasing hormone antagonist: how good is the new hope? AB - Gonadotropin-releasing hormone agonists have been widely used to prevent luteinizing hormone surges during controlled ovarian stimulation in assisted reproductive technologies. Treatment with gonadotropin-releasing hormone agonists of uterine myoma, endometriosis and some hormone-dependent cancers, such as breast, ovarian, endometrial and prostate cancer, also seems to have a beneficial effect. Gonadotropin-releasing hormone agonists have the disadvantage of inducing an initial stimulatory effect on gonadotropin secretion, necessitating 2-3 weeks before pituitary desensitization is achieved. Gonadotropin-releasing hormone antagonists, on the contrary, cause an immediate inhibition of gonadotropin secretion by competitive blocking of pituitary gonadotropin-releasing hormone receptors. Some advantages of their clinical use in controlled ovarian stimulation have already been demonstrated. Randomized comparative studies are needed to investigate their benefit over gonadotropin-releasing hormone antagonists for myoma and hormone-related disorders. PMID- 11396646 TI - Umbilical vein thrombosis as a possible cause of perinatal morbidity or mortality: report of two cases. AB - Significant occlusion of the lumen of the umbilical vein by thrombus was observed in 2 patients with unexplained intrauterine fetal death (IUFD)/fetal distress. Although a normal non-stress testing result was obtained 7 days prior to IUFD in one patient, IUFD was noted during regular antenatal care at 39 weeks of gestation; intrapartum abrupt onset of deceleration in fetal heart rate pattern was observed at 40 weeks of gestation, lasting 14 min until vacuum extractor assisted delivery in the other patient. Umbilical vein thrombosis was considered contributory to IUFD and the abrupt deterioration in fetal heart rate pattern in these 2 patients. Histological examination of the umbilical cord is thus important in unexplained fetal death/fetal distress. PMID- 11396648 TI - Recent progress in oocyte and ovarian tissue cryopreservation and transplantation. AB - Ovarian tissue cryopreservation and transplantation is an experimental technique that has been developed to sustain the reproductive function of women and children who are faced with sterilizing chemotherapy, radiotherapy, or radical reproductive surgery. Oocyte cryopreservation, on the other hand, is less feasible in the context of cancer because there is usually inadequate time to complete an ovarian stimulation cycle. The main promise of oocyte cryopreservation is that it offers an alternative when embryo freezing is not possible for technical, regulatory, or religious reasons. Oocyte freezing is more suitable for a single woman when the concern is age-related decline in fecundity. There have been significant scientific advances in the field of cryopreservation of ovarian tissue and oocytes, especially within the past few years. Ovarian function has been reported after the first cases of ovarian transplantation, and the number of pregnancies from cryopreserved oocytes has grown. Ovarian tissue and oocyte freezing can now be recommended in a carefully selected group of patients, provided that these options are offered under protocols that are approved by an institutional review board. PMID- 11396649 TI - Molecular markers of implantation: clinical implications. AB - The endometrium has been conventionally studied using histologic criteria. Our understanding of endometrial physiology has been advanced tremendously by research into the molecules that mediate its development and function. These molecules demonstrate a dynamic expression pattern through the menstrual cycle and have been implicated in endometrial growth, differentiation, and receptivity. These molecules include secreted proteins (endometrial bleeding-associated factor, glycodelin-A, insulin-like growth factor binding protein-1), cell-surface receptors (integrins), and nuclear transcription factors (HOXA10 and HOXA11). The homeobox genes Hoxa10 and Hoxa11 are necessary for implantation because mice with mutations in these genes exhibit a failure of implantation. HOXA10 and HOXA11 have been shown to be important for implantation in humans as well. Knowledge of endometrial molecular dynamics may now be used to enhance our ability to diagnose implantation defects. It may soon be possible to treat individual molecular defects by protein supplementation or gene therapy. PMID- 11396650 TI - Endometriosis and assisted reproductive technologies: are outcomes affected? AB - The preponderance of recent data suggests that endometriosis does not adversely affect in-vitro fertilization pregnancy rates. However, many studies demonstrate impaired oocyte quality, decreased fertilization, and compromised implantation rates. Such findings give insight into the mechanisms by which endometriosis may impact on fertility, and provide clues as how to focus assisted reproductive technologies in order to overcome these deficiencies. Specifically, extended downregulation protocols, ample use of gonadotropins for ovarian stimulation, and conservative management of endometriomas have all been suggested as means to optimize in-vitro fertilization outcomes for women with endometriosis. PMID- 11396651 TI - Hydrosalpinx and assisted reproduction: options and rationale for treatment. AB - Salpingoscopy during laparoscopy yields the best prognosis in patients with hydrosalpinx. It has been demonstrated that in-vitro fertilization (IVF) patients with hydrosalpinx have decreased pregnancy rates as compared with control individuals. If a patient with hydrosalpinx is to be treated with IVF, then the communication between the uterine tube and the uterine cavity should be blocked via salpingectomy or proximal tubal ligation, with or without distal tubal fenestration. This is because there is evidence that hydrosalpinx, especially when it is bilateral and visible by ultrasonography, impacts negatively on pregnancy and implantation rates after IVF cycles. PMID- 11396652 TI - Autoimmune factors in reproductive failure. AB - The present review highlights recent studies that investigated the possible influences of autoimmune factors in reproductive success or failure. These factors include antiphospholipid antibodies, antithyroid antibodies, antinuclear antibodies, antisperm antibodies, and antiovarian antibodies. The majority of recent work has focused on these potential autoimmune factors; however, controversy still exists over indicated testing and treatment options. An association of antiphospholipid antibodies and recurrent pregnancy loss has been established, and treatment with subcutaneous heparin appears most efficacious. Other autoimmune factors are under investigation as markers of in-vitro fertilization failure. Limited data from treatment trials are presented. PMID- 11396653 TI - Reflections on the number of embryos to replace at the dawn of the 21st century. AB - The present review examines recent publications concerned with the issue of the number of embryos to replace in an in-vitro fertilization cycle, and concentrates on the proposed selection methods that may allow us to transfer one or two embryos. A number of techniques that improve our selection criteria for deciding which is the most viable embryo are discussed, including blastocyst culture. It is proposed that the next step in assessing which embryos to transfer is to combine these techniques by performing assessment at the pronuclear stage, cleavage stages, and then after development to the blastocyst stage, when the number of embryos is sufficient. In so doing our embryo selection processes will be more critical, allowing us to limit the number of embryos replaced without detriment to pregnancy rates. PMID- 11396654 TI - Blastocyst transfer: does it really affect the outcome? AB - Blastocyst transfer is of great interest to most assisted reproductive technology clinics. The ability to select the best embryos to transfer in order to increase pregnancy rates, and to replace lower numbers of embryos to reduce the likelihood of an unwanted multiple pregnancy have both been realized. The optimal method to prolong embryo culture up to day 5 or 6 is still to be identified, although present techniques offer acceptable blastocyst development rates. Freezing of supernumerary blastocysts is still a matter of discussion, as adequate results are still awaited. PMID- 11396655 TI - Assisted conception: what does the future hold? PMID- 11396656 TI - The genetic basis of endometriosis. AB - Family studies have long suggested a role for genetic factors in the aetiology of endometriosis. The influence of genes on disease development has mainly been researched independently of environmental factors, yet their interaction must play an important role. Greater exposure to retrograde menstruation and oestrogen is likely to increase the risk of endometriosis; toxic compounds such as dioxin may increase the risk, although the only direct evidence has come from primate studies. Previous association studies implicated GALT (a gene involved in galactose metabolism), and GSTM1 and NAT2 (genes encoding for the detoxification enzymes) as possible disease susceptibility genes. Recent findings have added to the evidence for the involvement of GSTM1 and NAT2, but have cast doubt on the role of GALT. However, the design of many genetic and epidemiological studies has been inadequate with respect to sample size, consistency in phenotype definition, and the choice of control populations. These features are likely to influence results, and could partly explain the lack of consistency in the findings. Future studies should use a consistent disease definition and be of appropriate epidemiological design. PMID- 11396657 TI - Sex, infertility and the molecular biology of the androgen receptor. AB - Mutations that totally disrupt androgen receptor function cause the well known testicular feminizing syndrome or complete androgen insensitivity syndrome, wherein a 46 XY individual is completely feminized at birth. Recently it has been increasingly obvious that androgen receptor mutations not only result in the complete androgen insensitivity syndrome, but can cause a wide spectrum of milder insensitivity syndromes including ambiguous genitalia in newborn infants, and 'idiopathic' male infertility in otherwise normal males. Characterization of the molecular and structural mechanisms of androgen receptor dysfunction in these cases has led to directed hormonal therapy. Thus the differential response of a Met807Thr mutant androgen receptor to dihydrotestosterone but not testosterone, have been used to restore male genital development in an infant with partial AIS. Of greater significance, because they affect larger numbers of patients, are the mutations and polymorphisms that result in depressed spermatogenesis and male infertility in phenotypic males. Studies involving Singaporean, Australian, North American and Japanese subjects have established that increases in length of a trinucleotide repeat (CAG) tract, encoding a polyglutamine stretch in the transactivation domain of the androgen receptor, are associated with increased risk of defective spermatogenesis and undermasculinization. Independent of the CAG repeats, missense amino-acid substitutions in the ligand-binding domain, involving residues 727, 798 and 886 cause infertility through a novel mechanism. Pathogenicity is transmitted, not through defective ligand binding, but through defective protein-protein interactions between receptor domains and coactivator proteins that are essential for gene transcription. Elucidation of the molecular and structural basis of androgen receptor dysfunction in these cases allows precise genetic counseling and can lead to the design of rational hormonal therapy. PMID- 11396658 TI - Obesity, polycystic ovary syndrome and anovulation--how are they interrelated? AB - Polycystic ovary syndrome remains a mystery with respect to its aetiology, consequences and management. There is an increasing emphasis on its familial origins although the genes involved are unknown. There are significant changes in risk profiles for cardiovascular disease but no clear clinical evidence of morbidity or mortality. There is evidence of improvement in insulin resistance or insulin sensitivity drugs but uncertainty about improvement in pregnancy outcomes. This review looks at the latest advances in understanding polycystic ovary syndrome, particularly in obesity. PMID- 11396659 TI - Ovarian hyperstimulation syndrome: an endocrinopathy? AB - Ovarian hyperstimulation syndrome remains a significant but incompletely understood complication of ovarian stimulation. Evidence has accumulated regarding a role for various cytokines, in particular vascular endothelial growth factor, in its occurrence. However, the pathogenesis of ovarian hyperstimulation syndrome is likely to be complex and may involve a network of interacting cytokines and endocrine factors. Recent studies suggest links between specific cytokines and specific functional abnormalities in severe ovarian hyperstimulation syndrome. Two clinical forms of ovarian hyperstimulation syndrome may be distinguished, based on time of onset, with implications for the prediction and prevention of ovarian hyperstimulation syndrome in clinical practice. PMID- 11396661 TI - Bibliography. Current world literature. Fertility. PMID- 11396660 TI - Endometrial effects of etonogestrel (Implanon) contraceptive implant. AB - The subdermal contraceptive implant etonogestrel (Implanon; NV Organon International, Oss, The Netherlands) exerts complex effects on the endometrium. These include direct effects on the endometrium through endometrial progestin target sites, and indirect effects through suppression of the hypothalamic pituitary-ovarian axis. The resulting effects are categorized by alterations in endometrial histology, endometrial thickness, dysmenorrhoea and menstrual bleeding pattern. The exact mechanism of action of progestins on the endometrium has not been determined. The contraceptive effect in Implanon users is mainly due to inhibition of ovulation. Current research is concentrating on the potential of the progestin implant to modify endometrial vascular, angiogenic, steroid receptor and proto-oncogene function. These processes may be involved in the causation of progestin-induced breakthrough bleeding. PMID- 11396662 TI - Bibliography. Current world literature. Reproductive endocrinology. PMID- 11396663 TI - Now Gisborne Hospital. PMID- 11396664 TI - Changing risk behaviours for non-communicable disease in New Zealand working men- is workplace intervention effective? AB - AIMS: To evaluate the effectiveness of a health promotion programme targeting dietary behaviours and physical activity among male hourly-paid workers and to explore demographic and attitudinal influences on dietary patterns at baseline. METHODS: A controlled field trial compared workers at one intervention and one control worksite. The intervention comprised nutrition displays in the cafeteria and monthly 30-minute workshops for six months. Key outcome measures at six and twelve-months were self-reported dietary and lifestyle behaviours, nutrition knowledge, body mass index (BMI), waist circumference and blood pressure. RESULTS: 132 men at the intervention site and 121 men at the control site participated in the study and a high retention rate (94% at 6-months and 89% at 12-months) was achieved. At baseline, 40% of the total sample (253) were obese, 30% had elevated blood pressure, 59% indicated an excessive fat intake and 92% did not meet the recommended vegetable and fruit intake. The intervention reduced fat intake, increased vegetable intake and physical activity, improved nutrition knowledge and reduced systolic blood pressure when compared to the control site. There was no difference in change in mean BMI or waist circumference. Reduction in BMI was associated with reduction in fat intake. DISCUSSION: Low intensity workplace intervention can significantly improve reported health behaviours and nutrition knowledge although the impact on more objective measures of risk was variable. A longer duration or more intensive intervention may be required to achieve further reduction in risk factors. PMID- 11396665 TI - Cost-effectiveness of spinal cord stimulation in patients with intractable angina. AB - AIM: To review the cost of healthcare utilisation by patients suffering from intractable angina, unsuitable for coronary revascularisation, before and after treatment with spinal cord stimulation. METHODS: Data were collected for eight patients treated for intractable angina with spinal cord stimulation at Green Lane Hospital before April 1999. Information on consumption of specified medica resources for the twelve months preceding implantation, the implantation period, and the twelve months following implantation was collected. Where available, data were also collected for the eighteen months preceding and following treatment. RESULTS: In six patients successful permanent stimulation was established; in two it proved technically impossible to implant a stimulator. The six patients with successful stimulation spent fewer days in hospital (p=0.028) and consumed fewer resources (p=0.046) following implantation than in the period before implantation. The two patients for whom spinal cord stimulation was unsuccessful spent more days in hospital and consumed more resources in the twelve months following, than in the twelve months preceding attempted implantation. Extrapolation of data for all eight patients suggests that, on average, the cost of implanting a spinal cord stimulator will be recovered in approximately fifteen months. CONCLUSION: Spinal cord stimulation is a cost-effective treatment for intractable angina pectoris. PMID- 11396666 TI - Stereotactic radiosurgery in New Zealand. AB - AIMS: To explain the use of Stereotactic Radiosurgery for intracranial lesions and report Dunedin Hospital's early experience with this treatment. METHODS: Review of a prospective computer database and departmental clinical files. RESULTS: 74 patients underwent 78 radiosurgical procedures between 30 July 1994 to 18 December 1997. 28 patients with arteriovenous malformations were treated with an obliteration rate of 82% (95% CI: 48% to 98%) at two years. Seventeen vestibular schwannomas (acoustic neuroma's) were treated, with follow-up magnetic resonance imaging available in eleven in whom there was no tumour progression after a mean period of twelve months. There was preservation of some hearing in all patients not already deaf, but one developed a new facial palsy and another had worsening palsy as late side effects. Other tumours, including selected metastases, gliomas and skull base tumours have been treated in smaller numbers. CONCLUSION: Rates of arteriovenous malformation obliteration, vestibular schwannoma control, and side effects of radiosurgery in Dunedin are comparable to those reported in other uncontrolled series. Radiosurgery is quick and has a low procedure-related morbidity but does have important limitations and delayed side effects, which means the decision to treat needs to be based on thorough multidisciplinary review. PMID- 11396667 TI - Rehabilitation that works--vocational outcomes following rehabilitation for occupational musculoskeletal pain. AB - AIMS: To describe the short term vocational outcome for accident compensation claimaints with disabling musculoskeletal pain following a comprehensive, interdisciplinary rehabilitation programme. METHODS: A telephone follow-up audit of clients who had undertaken a rehabilitation programme characterised by a cognitive-behavioural approach with self-management, reconditioning, vocational rehabilitation and psychological pain management. RESULTS: Of 62 clients who had undergone a rehabilitation programme, we obtained follow-up information on 49 (79%). These were predominantly male, aged in their mid 30s, manual workers with low back pain and a median sick leave of twelve months. At a median of five months a vocational success was achieved in 75%: working full time (47%), part time (12%) or actively looking for work (16%). Of those in work, 48% went back to the same job, 7% went back to the same job but with a different employer and 15% went to a different job that used the same skills. Logistic regression analysis showed that duration of work disability was the major predictor of vocational success (OR 0.36, 95% CI 0.18 to 0.78, for a difference of twelve months). CONCLUSION: Despite the uncontrolled nature of these results, it is likely that the rehabilitation programme had a significant impact in getting compensation claimants back to work. Only a minority require substantive retraining and early intervention is associated with a better outcome. PMID- 11396668 TI - Prescribing the pill: obligations to health professionals. PMID- 11396669 TI - The RUB. Risk of Unacceptable Badness. PMID- 11396670 TI - What does the human genome project mean for medicine? PMID- 11396671 TI - Mandatory reporting of incompetence. PMID- 11396672 TI - ACE inhibitors for hypertension--again. PMID- 11396673 TI - Dengue fever in returned travellers. PMID- 11396674 TI - Kii ALS dementia. AB - Epidemiological surveys in the foci of ALS of the Kii Peninsula of Japan started in the early 1960s. Continuous surveys conducted for decades revealed that there have been two foci in the Kii Peninsula: one in Kozagawa in the southern part, and the other in Hobara in the south-east. Clinically, ALS patients of the Kii foci occasionally showed parkinsonian features or dementia that have not been reported in the sporadic form of ALS. Neuropathologically, numerous NFT that are identical to those of Alzheimer's disease were observed in the cerebral cortex and in the brainstem nuclei. To elucidate the etiopathogenesis of this unique form of ALS, an analysis was conducted of the environment in the focus areas and of the specimens from the patients with ALS. It was hypothesized that the long exposure of these environments to low calcium and magnesium, and an excess of aluminum and manganese in the drinking water and the soil, might lead to the deposition of some trace elements in the CNS, eventually causing neuronal degeneration and death. PMID- 11396675 TI - Sources of neuronal material for implantation. AB - The adult brain is an organ that does not have the natural ability to replace cells that have been lost through damage. Possible human interventions to rectify this situation include transplanting either developing neural tissue into the damaged host brain or transplantation of neural stem cells (cells that have the capacity to proliferate into neural cells and self-replicate) into the damaged area. Fetal or embryonic stem cells can be extracted and differentiated in vitro into the specific desired progeny (e.g. neurons). The neuronal stem cells themselves can be extracted from fetuses and multiplied in culture and then transplanted into the damaged brain. There is the possibility of dedifferentiation, in which cells of one type can be converted into a different cell type; for example, a differentiated blood cell could be de-differentiated back to its own hemopoietic stem cell and that stem cell could be converted into a neuronal stem cell which could then be differentiated into a neuron. It is probable that methods of generating large numbers of committed stem cells to treat conditions such as Alzheimer's disease will soon be increasingly common. PMID- 11396676 TI - Nicastrin, a key regulator of presenilin function, is expressed constitutively in human neural cell lines. AB - Nicastrin acts as a key regulator for presenilin (PS)-mediated gamma-secretase cleavage of beta-amyloid precursor protein by forming a functional complex with PS1 and PS2. Both TNF-alpha and IL-1, aberrantly produced by activated microglia and astrocytes, play a role in amyloidogenesis and neurodegeneration in the brains of Alzheimer's disease (AD) patients, while BDNF synthesized chiefly by neurons has been found to be substantially reduced in AD brains. To investigate the constitutive and cytokine/neurotrophic factor-regulated expression of nicastrin in human neural cells, its mRNA levels were studied by RT-PCR and northern blot analysis in SK-N-SH neuroblastoma cells, IMR-32 neuroblastoma cells, U-373MG astrocytoma cells, and NTera2 teratocarcinoma-derived differentiated neurons (NTera2-N) following exposure to TNF-alpha, IL-1beta, BDNF, dibutyryl cyclic AMP, or phorbol 12-myristate 13-acetate. Nicastrin mRNA expression was identified in all human neural and nonneural cell lines and tissues examined. The levels of nicastrin mRNA, however, were unaltered in SK-N SH, IMR-32, U-373MG, and NTera2-N cells by exposure to the factors tested, and unchanged in NTera2 cells during retinoic acid-induced neuronal differentiation. These results indicate that nicastrin mRNA is expressed constitutively in human neural cell lines, where its expression is not regulated at the transcriptional level by a battery of cytokines and growth/differentiation factors which are supposed to be involved in amyloidogenesis, neurodegeneration or neuroprotection in AD brains. PMID- 11396677 TI - Immunohistochemical expression and pathogenesis of BLM in the human brain and visceral organs. AB - Bloom syndrome (BS) involves the clinical features of telangiectatic erythema, immunodeficiency, and an increased risk for cancer. In order to clarify the pathogenetic significance of the responsible gene, BLM, which encodes a protein possessing homology to Escherichia coli RecQ helicase, the immunohistochemistry of BLM was examined in human brains and visceral organs from fetuses to adults and an adult with BS, using anti-BLM antibodies. Purkinje cells exhibited positive BLM immunoreactivity from 21 gestational weeks (GW), which transiently increased at approximately 40 GW. Neurons of the pontine tegmentum were immunolabeled from the early fetal period. In visceral organs, positive BLM immunoreactivity was observed in the Hassal corpuscles in the thymus from 24 GW, in beta-cells in the Langerhans islets of the pancreas from 36 GW, and in sperm cells and sperms of the testes from 11 years of age. But in a patient with BS, it was negative in the pancreas and testis tissues examined. The characteristic effect of BLM on specific cells in different periods suggests that the BLM gene product is closely related to neuronal development as well as immune, insulin secretory and sperm functions, which appear in different periods, and disorders of which are major symptoms of BS. PMID- 11396678 TI - JC virus large T protein transforms rodent cells but is not involved in human medulloblastoma. AB - JC virus (JCV) together with Simian virus 40 (SV40) and BK virus (BKV), belong to the polyomavirus group and these viruses are neuro-oncogenic to rodents by expression of large T antigen (LT), which binds to cellular p53 and pRB thus reducing the anticancer potential of the cell. The function of LT has not been clarified because small t antigen (st) is transcribed from the same start codon as the overlapping reading frame of LT, and is translated as a different protein with the same N-terminal residues (1-81 amino acids) by a splice-site variant of mRNA. To elucidate the function of LT without st, we constructed plasmids that express LT only by deleting the splicing region including the C-terminus of st, and consequently stable cell lines were established that express only JCLT, SV40LT and BKLT. The growth rates of these cells were examined in colonies on soft agar and it was found that LT alone has a transforming capacity; the order of efficiency being SV40LT, BKLT and JCLT. In addition, to verify the involvement of JCV in human medulloblastoma, eight cases of medulloblastoma, six cases of frozen material and five cases of paraffin-embedded tissues which included three cases of frozen tissues, were examined. PCR assay, genomic Southern blotting, and in situ hybridization were applied to detect the JCV genome, and LT and st were examined by immunohistochemistry; the results were compared with JCV-infected tissues as a positive control. All methods failed to detect not only JCV genome but also LT protein in medulloblastoma and it was concluded that JCV LT has transforming activities in rodent cells, but is not related to human medulloblastoma. PMID- 11396679 TI - Severe brain atrophy in a case of thalamic variant of sporadic CJD with plaque like PrP deposition. AB - A 30-year-old woman presented with ataxic gait and progressive mental deterioration, and 3 years later developed myoclonus in the limbs. Subsequently, she lapsed into an akinetic state and died more than 6 years after the onset of disease. The brain weighed 670 g, and preferential degeneration was found in the medial thalamus and the inferior olivary nucleus. In the cerebrum and cerebellum, gliosis and neuronal depletion were only mild and disintegration of the parenchymal structures was inconspicuous, despite pronounced atrophy. The patient had methionine homozygosity at codon 129 of the PrP gene and protease-resistant PrP type 2 in the brain. On PrP immunostaining, plaque-like deposits were detected in the cerebral and cerebellar cortices. Severe brain atrophy such as in the present case has never been described in the thalamic variant of sporadic CJD. PMID- 11396681 TI - Immunological mechanisms involved in experimental peptide immunotherapy of T-cell mediated diseases. AB - Current therapies for autoimmune diseases and allergy involve general immune suppression. However, the ideal therapy should specifically eliminate or modulate the (auto)pathogenic immune response or, alternatively, it should reinforce the regulatory response, without affecting the overall function of the immune system. This could be achieved by antigen-specific immunotherapy. Antigen-specific immunotherapy has received ample attention in the last years, and several clinical trials attempting to treat autoimmune diseases or allergy through the induction of antigen-specific tolerance or immune deviation have been conducted, albeit with varying success. Recent advances in our understanding of peripheral tolerance, regulatory T cells, and routes of antigen administration have resulted in better insight into the different working mechanisms and potential target molecules of antigen-specific immunotherapy. The experimental animal models and new technological developments force the pace in the development of these immunotherapies. The current review addresses several aspects of antigen-specific immunotherapies and focuses on the mechanisms of the different approaches in experimental autoimmune and allergy models. PMID- 11396682 TI - Expression of inhibitory receptors for MHC class I molecules on T cells. AB - Inhibitory receptors (IRs) specific for MHC class I molecules and originally described on natural killer (NK) cells are also expressed on a fraction of peripheral T cells. The presence of these receptors on T cells is poorly understood. In this review, the different antigen specificities described to date for IR+ T cells and the expression pattern of these receptors on T cells are analyzed. This analysis indicates that the population of T cells defined by IR expression is heterogeneous and that different IRs (or families of IRs) may play different roles in T-cell biology. PMID- 11396680 TI - Autoimmunity and the immunotherapy of cancer: targeting the "self" to destroy the "other". AB - It is increasingly clear that immunity to "self"-antigens may result in tumor destruction in mouse and man. But which antigens should be targeted with therapeutic cancer vaccines? In the case of melanoma, recognition of melanocyte differentiation antigens (MDA) can be associated with autoimmune depigmentation (vitiligo). We propose that intersection of protein transport to melanosomes and endosomes allows for the loading of MDA-derived peptides on MHC class II molecules, resulting in the activation of MDA-specific CD4+ "helper" T cells that aid the induction of melanoma-specific CD8+ T cells. Thus, the immunogenicity of MDA may be a consequence of their unique cell biology. Studies of MDA-based vaccines can provide new insight into the development of more effective cancer vaccines. PMID- 11396683 TI - Mechanisms of IgE elevation in HIV-1 infection. AB - Serum IgE levels are high in adults and children with HIV-1 infection and could be a marker of poor prognosis. Allergic reactions and adverse reactions to drugs also tend to increase in HIV-1-infected individuals. An imbalance between a "T(H)1-like" and a "T(H)2-like" cytokine profile has been documented in HIV-1 infection. We have demonstrated that HIV-1 gp 120 from different clades is a stimulus for histamine and cytokine (IL-4 and IL-13) release from basophils. Gp 120 acts as a viral superantigen, interacting with the V(H)3 region of IgE to induce mediator release from human Fc epsilonRI+ cells. Human basophils and mast cells express the chemokine receptor CCR3, which binds the chemokines eotaxin and RANTES. By interacting with the CCR3 receptor on Fc epsilonRI+ cells, HIV-I Tat protein is a potent chemoattractant for human basophils and lung mast cells. Preincubation of basophils with Tat protein upregulates mRNA CCR3 and the surface expression of this chemokine receptor. Tat also induces IL-4 and IL-13 release from basophils. Extracellular Tat can influence the directional migration of human Fc epsilonRI+ cells, the expression of chemokine receptor CCR3, and the release of T(H)2 cytokines. Our results indicate two novel mechanisms by which two HIV-1 proteins, gp120 and Tat, trigger the release of cytokines critical for T(H)2 polarization from human Fc epsilonRI+ cells. PMID- 11396684 TI - High-performance liquid chromatography coupled with chemiluminescence nitrogen detection for the study of ethoxyquin antioxidant and related organic bases. AB - The Chemiluminescent Nitrogen Detector (CLND) for use with high-performance liquid chromatography (HPLC) allows for the low-level detection of nitrogen containing compounds with simple quantitation. The nitrogen selective detector's equimolar response (i.e., equal response for nitrogen independent of its chemical environment) allows for any nitrogen-containing compound to be quantitated as long as the number of nitrogens are known. The HPLC-CLND provides a new detection method for analytes that are not available in large quantities or have unknown chemical or physical characteristics such as oxidation products, metabolites, or impurities. Ethoxyquin is a primary antioxidant that is used to preserve many food products and animal feeds. HPLC-CLND is used in the study of the oxidation products of ethoxyquin because limited quantities of these compounds are available and subsequent calibration curves are difficult to maintain. HPLC-CLND as a new method of detection has been evaluated for its equimolarity of response, linear range, limit of detection, and limit of quantitation. PMID- 11396685 TI - Direct thermal desorption in the analysis of cheese volatiles by gas chromatography and gas chromatography-mass spectrometry: comparison with simultaneous distillation-extraction and dynamic headspace. AB - Direct thermal desorption (DTD) has been used as a technique for extracting volatile components of cheese as a preliminary step to their gas chromatographic (GC) analysis. In this study, it is applied to different cheese varieties: Camembert, blue, Chaumes, and La Serena. Volatiles are also extracted using other techniques such as simultaneous distillation-extraction and dynamic headspace. Separation and identification of the cheese components are carried out by GC-mass spectrometry. Approximately 100 compounds are detected in the examined cheeses. The described results show that DTD is fast, simple, and easy to automate; requires only a small amount of sample (approximately 50 mg); and affords quantitative information about the main groups of compounds present in cheeses. PMID- 11396686 TI - Behavior of haloperidol and various phenothiazines on several alkyl bonded phases. AB - Haloperidol and phenothiazines are present in psychiatrical treatments. An analysis in body fluids is tedious because of the presence of demethylated (DM) derivatives of phenothiazines. The behavior of some interfering solutes on alkyl bonded phases has been studied. Phenothiazines and DM derivatives exhibit a very similar behavior with a binary eluent (phosphate buffer-acetonitrile), which precludes an optimization with this system. When a ternary phase is used (phosphate buffer-acetonitrile-methanol), haloperidol and reduced haloperidol behave differently as compared with phenothiazines. In this mode it is possible to unambiguously detect haloperidol that would otherwise interfere. Phenothiazine peaks are characterized by a large tailing. An interesting feature is the comparison between cyclohexyl bonded and octadecyl bonded phases, the former being much more efficient. PMID- 11396687 TI - Determination of carboxylic acids, carbohydrates, glycerol, ethanol, and 5-HMF in beer by high-performance liquid chromatography and UV-refractive index double detection. AB - A high-performance liquid chromatographic method is proposed for the simultaneous separation of main carboxylic acids, carbohydrates, ethanol, glycerol, and 5-HMF in beer by direct injection. A column packed with a sulfonated divinyl benzene styrene copolymer and an isocratic elution with 0.0045N sulfuric acid and acetonitrile (6%, v/v) are employed. UV and refractive index detectors connected in series are also used to reduce the matrix interference of phenolic compounds. In conditions described, nine compounds are quantitated in a single chromatographic run without any pretreatment except for sample dilution and filtration before injection. Precision, accuracy, linearity of response, limit of detection, and limit of quantitation are also evaluated for each compound. Satisfactory results are obtained to justify the application of this method to all phases of beer production for process and quality control. PMID- 11396688 TI - Quantitation of ergosterol in river sediment by liquid chromatography. AB - The ergosterol content of a river sediment can be used as an indicator of fungal activity. A method is developed for the extraction and determination of ergosterol in river sediment as part of a study to assess the correlation between fungal activity and biodegradation of pyrene, which is an environmental pollutant. This method is based on saponification and the liquid-liquid extraction of ergosterol by ethyl acetate. Quantitation and detection are performed isocratically by liquid chromatography on a 5-microm Hypersil C18 column with methanol-acetonitrile (80:20, v/v) as the mobile phase and detection at 282 nm. The detection limit is 50 ng/mL ergosterol, which is equivalent to 0.1 microg/g. The recovery of ergosterol at a concentration level in the range of 2 to 12 microg/mL is 91.7% +/- 3.1% without interferences. This method is applied in order to successfully quantitate the ergosterol content in a river sediment with or without a fungus supply. PMID- 11396689 TI - Amino acid, fatty acid, and carbohydrate content of Artocarpus altilis (breadfruit). AB - A study is conducted to determine the amino acid, fatty acid, and carbohydrate content of breadfruit using high-performance liquid chromatography (HPLC) and gas chromatography (GC). An HPLC method is used for the determination of amino acids and fatty acids in breadfruit. Representative amino acid samples are derivatized with phenylisothiocianate and the resulting phenylthiocarbamyl derivatives are separated on a reversed-phase column by gradient elution with a 0.05M ammonium acetate buffer and 0.01M ammonium acetate in acetonitrile-methanol-water (44:10:46, v/v). Representative fatty acid samples are derivatized with phenacyl bromide and the resulting fatty acid phenacyl esters are separated on a reversed phase column by gradient elution with acetonitrile and water. Amino acid and fatty acid derivatives are detected by ultraviolet detection at 254 nm. The analysis of the carbohydrates in breadfruit employs a GC method. Carbohydrates are derivatized using trimethylchlorosilane and hexamethyldisilazane to form trimethylsilyl ethers. Compounds in the samples are separated by the temperature programming of a GC using nitrogen as the carrier gas. Percent recoveries of amino acids, fatty acids, and carbohydrates are 72.5%, 68.2%, and 81.4%, respectively. The starch content of the breadfruit is 15.52 g/100 g fresh weight. PMID- 11396690 TI - Simultaneous determination of p-hydroxybenzaldehyde, p-hydroxybenzyl alcohol, 4 (beta-D-glucopyranosyloxy)-benzyl alcohol, and sugars in Gastrodia elata blume measured as their acetylated derivatives by GC-MS. AB - A method for the simultaneous separation and determination of the active constituents and three sugars in the roots of Gastrodia elata Blume (GE), which is used as a famous Chinese traditional herbal medicine, by gas chromatography mass spectrometry is established. The samples are acetylated with pyridine-acetic anhydride. The contents of 4-hydroxybenzaldehyde, 4-hydroxybenzyl alcohol (HA), fructose, glucose, 4-(beta-D-glucopyranosyloxy)-benzyl alcohol (GA), and sucrose in GE are 0.004%, 0.03%, 1.36%, 1.12%, 1.97%, and 4.25%, respectively, and the detection limits are 1.5, 3.0, 11.0, 5.0, 33.0, and 35.0 pg, respectively. The contents of HA and GA in the urine and brain of a mouse are also determined. This method is simple, reliable, and quick for the simultaneous determination of the active constituents and sugars in GE. PMID- 11396692 TI - Could you explain what is meant by the term rapid-analysis HPLC and give me some suggestions about when I might consider using it in optimizing a method? PMID- 11396691 TI - Collaborative study of EPA Method 317.0 for the determination of inorganic oxyhalide disinfection by-products in drinking water using ion chromatography with the addition of a postcolumn reagent for trace bromate analysis. AB - The development of the U.S. Environmental Protection Agency (EPA) Method 317.0 is initiated to provide a sufficiently sensitive and fundamental technique for the compliance monitoring of trace levels of bromate in drinking water. After a comparative evaluation of Method 317.0 and elimination of a chlorite interference, this method is tested by a collaborative study in order to determine the precision and bias of the method and evaluate its potential role as a future compliance-monitoring method for inorganic disinfection by-products (DBPs) and trace bromate. This technique provides a practical method for future compliance monitoring for all of the inorganic oxyhalide DBPs including trace concentrations of bromate. PMID- 11396693 TI - Dietary fiber and weight regulation. AB - The influence of dietary fiber on energy regulation remains controversial. This review summarizes published studies on the effects of dietary fiber on hunger, satiety, energy intake, and body composition in healthy individuals. Under conditions of fixed energy intake, the majority of studies indicate that an increase in either soluble or insoluble fiber intake increases postmeal satiety and decreases subsequent hunger. When energy intake is ad libitum, mean values for published studies indicate that consumption of an additional 14 g/day fiber for >2 days is associated with a 10% decrease in energy intake and body weight loss of 1.9 kg over 3.8 months. Furthermore, obese individuals may exhibit a greater suppression of energy intake and body weight loss (mean energy intake in all studies was reduced to 82% by higher fiber intake in overweight/obese people versus 94% in lean people; body weight loss was 2.4 kg versus 0.8 kg). These amounts are very similar to the mean changes in energy intake and body weight changes observed when dietary fat content is lowered from 38% to 24% of energy intake in controlled studies of nonobese and obese subjects. The observed changes in energy intake and body weight occur both when the fiber is from naturally high fiber foods and when it is from a fiber supplement. In view of the fact that mean dietary fiber intake in the United States is currently only 15 g/day (i.e., approximately half the American Heart Association recommendation of 25-30 g/day), efforts to increase dietary fiber in individuals consuming <25 g/day may help to decrease the currently high national prevalence of obesity. PMID- 11396694 TI - Iron and colorectal cancer risk: human studies. AB - Some reports have associated iron with cancer risk, particularly of the colorectum. This review will focus on the human studies that have investigated this association. Comparative studies were sought in which people with and without colorectal neoplastic lesions, either cancers or adenomatous polyps, were assessed for iron exposure. Iron exposure variables included dietary iron intake, iron vitamin supplementation, body iron stores as measured by ferritin or transferrin saturation, and gene status for hereditary hemochromatosis. Medline was searched for published reports using the key words iron, cancer, colon, rectum, ferritin, transferrin, and hemochromatosis. In addition, the Cochrane Library was searched for relevant studies and several authors were contacted to investigate their awareness of unpublished studies. Studies were categorized by study design and ranked for quality of innovation in design, sample size, and thoroughness of iron status ascertainment. Thirty-three studies were reviewed in 26 publications. Of the larger studies, approximately three-quarters supported the association of iron, in all three strata of exposure, with colorectal neoplasia risk. Because iron is broadly supplemented in the American diet, the benefits of iron supplementation need to be measured against the long-term risks of increased iron exposure, one of which may be increased risk of colorectal cancer. PMID- 11396695 TI - Whole grain intake and risk of ischemic stroke in women. AB - In a recent prospective study, whole grain intake was associated with a reduced risk of ischemic stroke. Refined grain, however, did not elicit such protection. Given the potential health benefits associated with whole grain diets, further exploration on the effects of whole grains on health and risk of chronic disease is warranted. PMID- 11396696 TI - High calcium intake blunts pregnancy-induced increases in maternal blood lead. AB - In pregnant women with low exposure to lead, high intakes of calcium (>2,000 mg/day) decreased the serum concentration of lead, which could potentially minimize fetal exposure to lead. This is twice the amount of calcium recommended for women during pregnancy and approaches the Upper Level for calcium of 2,500 mg/day. The mechanism by which high calcium intake blunts pregnancy-induced increases in maternal blood lead may involve decreased lead absorption in the intestine or decreased maternal bone resorption with subsequent release of lead. Either mechanism could decrease maternal blood concentrations of lead and potentially limit fetal accumulation of lead. PMID- 11396697 TI - Eat a variety of healthful foods: old advice with new support. AB - Although dietary guidelines have long advised eating a variety of foods, most nutrition research has focused on specific nutrients and their relationships to health and disease. The recent failure of several randomized trials to confirm results of observational studies linking specific nutrients to disease outcomes, and a growing understanding of the beneficial phytochemicals in the natural food supply, has redirected attention to the complexity and importance of the foods themselves. Studies identifying associations between diet quality and mortality are beginning to quantify the importance of eating a variety of healthful foods and are helping to refine that advice. PMID- 11396698 TI - The physiology and pathophysiology of iodine and the thyroid. PMID- 11396699 TI - Update on intrathyroidal iodine metabolism. AB - The thyroid concentrates iodide from the serum and oxidizes it at the apical membrane, attaching it to tyrosyl residues within thyroglobulin (Tg) to make diiodotyrosine and monoiodotyrosine. Major players in this process are Tg, thyroperoxidase (TPO), hydrogen peroxide, pendrin, and nicotinamide adenine dinucleotide phosphate (NADPH). Further action of TPO, hydrogen peroxide (H2O2), and iodinated Tg produce thyroxine (T4) and triiodothyronine (T3). Hormone containing Tg is stored in the follicular lumen, then processed, most commonly by micropinocytosis. The lysosomal enzymes cathepsins B, L, and D are active in Tg proteolysis. Tg digestion leaves T4 and T3 intact, to be released from the cell, while the 3,5'-diiodotyrosine (DIT) and 3-iodotyrosine (MIT) are retained and deiodinated for recycling within the thyroid. Some areas of especially active recent research include: (1) the role of molecular chaperones in directing properly folded TPO and Tg to the apical membrane; (2) details of proteolytic pathways; (3) modulation of iodine metabolism, not only by thyrotropin (TSH) but by iodine supply and by feedback effects of Tg, glutathione, and inhibitory elements in the N-terminal region of Tg; and (4) details of Tg structure and iodotyrosyl coupling. Despite general agreement on the major steps in intrathyroidal iodine metabolism, new details of mechanisms are constantly being uncovered and are greatly improving understanding of the overall process. PMID- 11396700 TI - Sodium iodide symporter in health and disease. AB - Radioiodine-concentrating activity in thyroid tissues has allowed the use of radioiodine as a diagnostic and therapeutic agent for patients with thyroid disorders such as well-differentiated thyroid cancer. However, some extrathyroidal tissues also take up radioiodine, contributing to unwanted side effects of radioiodine therapy. Now that the molecule that mediates radioiodine uptake, the sodium iodide symporter (NIS), has been cloned and characterized, it may be possible to develop novel strategies to differentially modulate NIS expression and/or activity, enhancing it in target tissues and impeding it in others. In addition to restoring NIS expression/activity to ensure sufficient radioiodine uptake for the diagnosis and treatment of advanced thyroid cancers, we envision that it may be possible to selectively increase or confer NIS expression/activity in tumors of nonthyroidal tissues to facilitate the use of radioiodine in their diagnosis and treatment. We also consider the molecular basis of thyroid and nonthyroid disorders that may be complicated by NIS deregulation. Finally, we explore the use of NIS as an imaging reporter gene to monitor the expression profile of the transgene in transgenic mouse animal models and in patients undergoing gene therapy clinical trials. PMID- 11396701 TI - Iodine and thyroid autoimmune disease in animal models. AB - Thyroid autoimmune diseases are complex, polygenic afflictions the penetrance of which is heavily dependent on various environmental influences. In their pathogenesis, an afferent stage (enhanced autoantigen presentation), a central stage (excessive expansion and maturation of autoreactive T and B cells), and an efferent stage (effects of autoreactive T cells and B cells on their targets) can be discerned. At each stage, a plethora of inborn, endogenous or exogenous factors is able to elicit the abnormalities characteristic of that stage, thus opening the gateway to thyroid autoimmunity. Iodine is an important exogenous modulating factor of the process. In general, iodine deficiency attenuates, while iodine excess accelerates autoimmune thyroiditis in autoimmune prone individuals. In nonautoimmune prone individuals, the effects of iodine are different. Here iodine deficiency precipitates a mild (physiological) form of thyroid autoimmune reactivity. Iodine excess stimulates thymus development. Iodine probably exerts these effects via interference in the various stages of the autoimmune process. In the afferent and efferent stage, iodine-induced alterations in thyrocyte metabolism and even necrosis most likely play a role. By contrast, in the central phase, iodine has direct effects on thymus development, the development and function of various immune cells (T cells, B cells macrophages and dendritic cells) and the antigenicity of thyroglobulin. PMID- 11396702 TI - Iodine deficiency in the world: where do we stand at the turn of the century? AB - Iodine deficiency is the leading cause of preventable mental retardation. Universal salt iodization (USI), calling for all salt used in agriculture, food processing, catering and household to be iodized, is the agreed strategy for achieving iodine sufficiency. This article reviews published information on programs for the sustainable elimination of the iodine deficiency disorders and reports new data on monitoring and impact of salt iodization programs at the population level. Currently, 68% of households from areas of the world with previous iodine deficiency have access to iodized salt, compared to less than 10% a decade ago. This great achievement, a public health success unprecedented in the field of noncommunicable diseases, must be better recognized by the health sector, including thyroidologists. On the other hand, the managers and sponsors of programs of iodized salt must appreciate the continuing need for greatly improved monitoring and quality control. For example, partnership evaluation of iodine nutrition using the ThyroMobil model in 35,223 schoolchildren at 378 sites of 28 countries has shown that many previously iodine deficient parts of the world now have median urinary iodine concentrations well above 300 microg/L, which is excessive and carries the risk of adverse health consequences. The elimination of iodine deficiency is within reach but major additional efforts are required to cover the whole population at risk and to ensure quality control and sustainability. PMID- 11396703 TI - The toxicology of iodate: a review of the literature. AB - Because it is more stable than iodide, most health authorities preferentially recommend iodate as an additive to salt for correcting iodine deficiency. Even though this results in a low exposure of at most 1,700 microg/d, doubts have recently been raised whether the safety of iodate has been adequately documented. In humans and rats, oral bioavailability of iodine from iodate is virtually equivalent to that from iodide. When given intravenously to rats, or when added to whole blood or tissue homogenates in vitro or to foodstuff, iodate is quantitatively reduced to iodide by nonenzymatic reactions, and thus becomes available to the body as iodide. Therefore, except perhaps for the gastrointestinal mucosa, exposure of tissues to iodate might be minimal. At much higher doses given intravenously (i.e., above 10 mg/kg), iodate is highly toxic to the retina. Ocular toxicity in humans has occurred only after exposure to doses of 600 to 1,200 mg per individual. Oral exposures of several animal species to high doses, exceeding the human intake from fortified salt by orders of magnitude, pointed to corrosive effects in the gastrointestinal tract, hemolysis, nephrotoxicity, and hepatic injury. The studies do not meet current standards of toxicity testing, mostly because they lacked toxicokinetic data and did not separate iodate-specific effects from the effects of an overdose of any form of iodine. With regard to tissue injury, however, the data indicate a negligible risk of the small oral long-term doses achieved with iodate-fortified salt. Genotoxicity and carcinogenicity data for iodate are scarce or nonexisting. The proven genotoxic and carcinogenic effects of bromate raise the possibility of analogous activities of iodate. However, iodate has a lower oxidative potential than bromate, and it did not induce the formation of oxidized bases in DNA under conditions in which bromate did, and it may therefore present a lower genotoxic and carcinogenic hazard. This assumption needs experimental confirmation by proper genotoxicity and carcinogenicity data. These in turn will have to be related to toxicokinetic studies, which take into account the potential reduction of iodate to iodide in food, in the intestinal lumen or mucosa, or eventually during the liver passage. PMID- 11396704 TI - Environmental iodine intake affects the type of nonmalignant thyroid disease. AB - The relationship between the iodine intake level of a population and the occurrence of thyroid diseases is U-shaped with an increase in risk from both low and high iodine intakes. Developmental brain disorders and endemic goiter caused by severe iodine deficiency may seriously deteriorate overall health status and economic performance of a population. Severe iodine deficiency with a median 24 hour urinary iodine excretion of the population below 25 microg needs immediate attention and correction. Less severe iodine deficiency with median urinary iodine excretion below 120 microg per 24 hours is associated with multinodular autonomous growth and function of the thyroid gland leading to goiter and hyperthyroidism in middle aged and elderly subjects. The lower the iodine intake, the earlier and more prominent are the abnormalities. At the other end of the spectrum, severely excessive iodine intake starting at median urinary iodine excretion levels around 800 microg per 24 hours is associated with a higher prevalence of thyroid hypofunction and goiter in children. A number of studies indicate that moderate and mild iodine excess (median urinary iodine >220 microg per 24 hours) are associated with a more frequent occurrence of hypothyroidism, especially in elderly subjects. The exact mechanism leading to this has not been clarified, and more studies are needed to define the limits of excessive iodine intake precisely. Due to the frequent occurrence of thyroid disorders, proper monitoring and control of the population iodine intake level is a cost-effective alternative to diagnosing, therapy and control of the many individual cases of thyroid diseases that might have been prevented. PMID- 11396705 TI - Pregnancy and iodine. AB - Hormonal changes and metabolic demands during pregnancy result in profound alterations in the biochemical parameters of thyroid function. For thyroid economy, the main events occurring during pregnancy are a marked increase in serum thyroxine-binding globulin levels; a marginal decrease in free hormone concentrations (in iodine-sufficient areas) that is significantly amplified when there is iodine restriction or overt iodine deficiency; a frequent trend toward a slight rise in basal thyrotropin (TSH) values between the first trimester and term; a transient stimulation of the maternal thyroid gland by elevated levels of human chorionic gonadotropin (hCG) resulting in a rise in free thyroid hormones and decrement in serum TSH concentrations during the first trimester; and finally, modifications of the peripheral metabolism of maternal thyroid hormones. Together, metabolic changes associated with the progression of gestation in its first half constitute a transient phase from preconception steady state to pregnancy steady state. In order to be met, these metabolic changes require an increased hormonal output by the maternal thyroid gland. Once the new equilibrium is reached, increased hormonal demands are maintained until term, probably through transplacental passage of maternal thyroid hormones and increased turnover of maternal thyroxine (T4), presumably under the influence of the placental (type 3) deiodinase. For healthy pregnant women with iodine sufficiency, the challenge of the maternal thyroid gland is to adjust the hormonal output in order to achieve the new equilibrium state, and thereafter maintain the equilibrium until term. In contrast, the metabolic adjustment cannot easily be reached during pregnancy when the functional capacity of the thyroid gland is impaired because of iodine deficiency. The ideal dietary allowance of iodine recommended by World Health Organization (WHO) is 200 microg of iodine per day for pregnant women. In conditions with iodine restriction, enhanced thyroidal stimulation is revealed by relative hypothyroxinernia and goitrogenesis. Goiters formed during gestation may only partially regress after parturition. Pregnancy, therefore, represents one of the environmental factors that may help explain the higher prevalence of goiter and thyroid disorders in women compared with men. An iodine-deficient status in the mother also leads to goiter formation in the progeny and neuropsycho-intellectual impairment in the offspring. When adequate iodine supplementation is given early during pregnancy, it allows for the correction and almost complete prevention of maternal and neonatal goitrogenesis. In summary, pregnancy is accompanied by profound alterations in the thyroid economy, resulting from a complex combination of factors specific to the pregnant state, which together concur to stimulate the maternal thyroid machinery. Increased thyroidal stimulation induces, in turn, a sequence of events leading from physiological adaptation of the thyroidal economy observed in healthy iodine sufficient pregnant women to pathological alterations affecting both thyroid function and the anatomical integrity of the thyroid gland, when gestation takes place in conditions with iodine restriction or deficiency: the more severe the iodine deficiency, the more obvious, frequent, and profound the potential maternal and fetal repercussions. PMID- 11396706 TI - Iodine and cancer. AB - Thyroid carcinomas are the most frequent endocrine malignancies. Among thyroid carcinomas the most frequent types are the differentiated forms (follicular, papillary or mixed papillary-follicular), whereas anaplastic thyroid carcinoma and medullary thyroid carcinomas are rare. Animal experiments have demonstrated a clear increase in incidence of thyroid epithelial cell carcinomas after prolonged iodine deficiency leading to a situation of the thyroid gland by thyrotropin and possibly other growth factors. However, the overall incidence of differentiated thyroid carcinoma is generally not considered to be influenced by the iodine intake of a population, whereas the distribution of the types of thyroid carcinoma seems to be related to the intake of iodine, with fewer of the more aggressive follicular and anaplastic carcinomas and more papillary carcinomas in iodine rich areas. Populations starting iodine prophylaxis demonstrate an increase in the ratio of papillary to follicular carcinoma. Because a population with higher iodine intake usually has fewer benign nodules in the thyroid gland and the incidence of thyroid carcinomas is similar to an iodine-deficient region, the diagnostic work-up of nodules in the thyroid gland may become affected. The incidence of other cancers, such as breast cancer, may be influenced by the iodine intake, but too few studies are available at present. The present article summarizes available data from both epidemiological studies, animal experiments, and basic gene transfection studies. The overall incidence for a relationship between iodine and cancer is poor and future studies are warranted. PMID- 11396707 TI - Iodine nutrition and the risk from radioactive iodine: a workshop report in the chernobyl long-term follow-up study. AB - The major fallout of radionuclides from the nuclear power station accident at Chernobyl on 26 April, 1986, occurred in regions of Ukraine and Belarus that are believed to be moderately deficient in dietary iodine. On 17 November, 2000, in conjunction with the Ukraine-Belarus-USA study of developing thyroid disease in a cohort of individuals exposed as children, a workshop was held to review what is known about iodine nutrition in the region, how this might influence the risk of thyroid tumor formation from radioiodine, and whether and how iodine nutrition should be monitored in this long-term project. This report is a summary of the workshop proceedings. Although no precise information about iodine intake in 1986 was found, the prevalence of mild goiter in the region's children suggested iodine deficiency and urinary iodine measurements begun in 1990 indicated that mild to moderate deficiency existed. Increased thyroid iodine uptake and increased thyroid size in 1986 resulting from iodine deficiency would have had counteracting influence on the thyroid radiation dose and knowledge of these parameters is required for dose reconstruction. More problematic is the possible role of iodine deficiency in the years following the accident. Theoretically, the resulting increase in thyroid cellular activity might increase the risk of tumorigenesis but experimental or clinical evidence supporting this hypothesis is meager or absent. Despite this limitation it was considered important to monitor iodine nutrition in the cohort subjects in relation to their place of residence and over time. Methods to accomplish this were discussed. PMID- 11396708 TI - Iodine excess and hyperthyroidism. AB - 150 microg iodine are daily required for thyroid hormone synthesis. The thyroid gland has intrinsic mechanisms that maintain normal thyroid function even in the presence of iodine excess. Large quantities of iodide are present in drugs, antiseptics, contrast media and food preservatives. Iodine induced hyperthyroidism is frequently observed in patients affected by euthyroid iodine deficient goiter when suddenly exposed to excess iodine. Possibly the presence of autonomous thyroid function permits the synthesis and release of excess quantities of thyroid hormones. The presence of thyroid autoimmunity in patients residing in iodine-insufficient areas who develop iodine-induced hyperthyroidism has not been unanimously observed. In iodine-sufficient areas, iodine-induced hyperthyroidism has been reported in euthyroid patients with previous thyroid diseases. Euthyroid patients previously treated with antithyroid drugs for Graves' disease are prone to develop iodine-induced hyperthyroidism. As well, excess iodine in hyperthyroid Graves' disease patients may reduce the effectiveness of the antithyroid drugs. Occasionally iodine-induced hyperthyroidism has been observed in euthyroid patients with a previous episode of post-partum thyroiditis, amiodarone destructive or type II thyrotoxicosis and recombinant interferon-alpha induced destructive thyrotoxicosis. Amiodarone administration may induce thyrotoxicosis. Two mechanisms are responsible for this condition. One is related to excess iodine released from the drug, approximately 9 mg of iodine following a daily dose of 300 mg amiodarone. This condition is an iodine-induced thyrotoxicosis or type I amiodarone-induced thyrotoxicosis. The other mechanism is due to the amiodarone molecule that induces a destruction of the thyroid follicles with a release of preformed hormones. This condition is called amiodarone-induced destructive thyrotoxicosis or type II thyrotoxicosis. Patients developing type I thyrotoxicosis in general have preexisting nodular goiter whereas those developing type II thyrotoxicosis have a normal thyroid gland. The latter group of patients, after recovering from the destructive process, may develop permanent hypothyroidism as the consequence of fibrosis of the gland. PMID- 11396709 TI - Iodine-Induced hypothyroidism. AB - Iodine is an essential element for thyroid hormone synthesis. The thyroid gland has the capacity and holds the machinery to handle the iodine efficiently when the availability of iodine becomes scarce, as well as when iodine is available in excessive quantities. The latter situation is handled by the thyroid by acutely inhibiting the organification of iodine, the so-called acute Wolff-Chaikoff effect, by a mechanism not well understood 52 years after the original description. It is proposed that iodopeptide(s) are formed that temporarily inhibit thyroid peroxidase (TPO) mRNA and protein synthesis and, therefore, thyroglobulin iodinations. The Wolff-Chaikoff effect is an effective means of rejecting the large quantities of iodide and therefore preventing the thyroid from synthesizing large quantities of thyroid hormones. The acute Wolff-Chaikoff effect lasts for few a days and then, through the so-called "escape" phenomenon, the organification of intrathyroidal iodide resumes and the normal synthesis of thyroxine (T4) and triiodothyronine (T3) returns. This is achieved by decreasing the intrathyroidal inorganic iodine concentration by down regulation of the sodium iodine symporter (NIS) and therefore permits the TPO-H202 system to resume normal activity. However, in a few apparently normal individuals, in newborns and fetuses, in some patients with chronic systemic diseases, euthyroid patients with autoimmune thyroiditis, and Graves' disease patients previously treated with radioimmunoassay (RAI), surgery or antithyroid drugs, the escape from the inhibitory effect of large doses of iodides is not achieved and clinical or subclinical hypothyroidism ensues. Iodide-induced hypothyroidism has also been observed in patients with a history of postpartum thyroiditis, in euthyroid patients after a previous episode of subacute thyroiditis, and in patients treated with recombinant interferon-alpha who developed transient thyroid dysfunction during interferon-a treatment. The hypothyroidism is transient and thyroid function returns to normal in 2 to 3 weeks after iodide withdrawal, but transient T4 replacement therapy may be required in some patients. The patients who develop transient iodine-induced hypothyroidism must be followed long term thereafter because many will develop permanent primary hypothyroidism. PMID- 11396710 TI - The various effects of amiodarone on thyroid function. AB - Amiodarone, a benzofuranic-derivative iodine-rich drug used mostly for tachyarrhythmias, often causes changes in the peripheral metabolism of thyroid hormones mainly due to the inhibition of 5'-deiodinase activity: an increase in serum thyroxine and reverse triiodothyronine, and a decrease in serum triiodothyronine concentrations. Overt thyroid dysfunction, either amiodarone induced thyrotoxicosis (AIT) or amiodarone-induced hypothyroidism (AIH), occurring in 14% to 18% of patients receiving long-term treatment, may develop both in apparently normal thyroid glands and in glands with preexisting abnormalities. AIH is mainly due to the failure to escape from the acute Wolff Chaikoff effect, and, in patients with thyroid autoimmune phenomena, to concomitant Hashimoto's thyroiditis. AIT is due to excess iodine-induced thyroid hormone synthesis (type I AIT) or to amiodarone-related destructive thyroiditis (type II AIT), although mixed forms often occur. Treatment of AIH consists of levothyroxine replacement therapy while continuing amiodarone therapy; alternatively, amiodarone can be discontinued, if possible, and the natural course toward euthyroidism can be accelerated by a short trial of potassium perchlorate. In type I AIT, the simultaneous administration of thionamides and potassium perchlorate is the treatment of choice, while in type II AIT steroids are the most useful therapeutic option. Mixed forms are best treated with a combination of thionamides, potassium perchlorate, and glucocorticoids. The low thyroidal 131I uptake usually makes radioiodine therapy not feasible, while thyroidectomy is a valid alternative in cases resistant to medical therapy. PMID- 11396711 TI - The unusual cartilaginous tissues of jawless craniates, cephalochordates and invertebrates. AB - A collagenous extracellular matrix was previously considered to be a requirement for classification of true cartilage. Data from the lamprey and hagfish now clearly indicate that both of these jawless craniates have extensive non collagenous, yet cartilaginous endoskeletons. Non-collagenous cartilages are present in the cephalochordates (amphioxus) and in the invertebrates, although collagen-containing cartilages also are found in the invertebrates. This review summarizes current knowledge of the morphological, biochemical and molecular characteristics of the unusual non-collagenous cartilages in jawless craniates and the cartilaginous tissues in amphioxus and invertebrates. A least two types of non-collagenous cartilage matrix proteins are found in both the hagfishes and the lampreys, all of which are resistant to digestion by cyanogen bromide (CNBr). Although all four of these matrices show some similarities with each other, suggesting a family of non-collagenous, elastin-like proteins, it is clear that the major matrix proteins of each are different. New morphological and biochemical information on the cartilaginous tissues in squid, horseshoe crab and amphioxus reveals the presence of CNBr-insoluble, non-collagenous matrix proteins, potentially extending the jawless craniate family of cartilaginous proteins into the invertebrates. Details of the evolutionary relationships between these non-collagenous matrix proteins and the significance of the occurrence of these proteins as the major components of the cartilaginous tissues of jawless craniates, amphioxus, horseshoe crab and squid, all of which are capable of producing a variety of collagens in other tissues, remain to be investigated. PMID- 11396712 TI - Intrinsic choroidal neurons in the duck eye receive sympathetic input: anatomical evidence for adrenergic modulation of nitrergic functions in the choroid. AB - Intrinsic choroidal neurons (ICN) in the duck eye form an intramural ganglionic plexus that may subserve complex integrative functions. A key feature of such ganglia is an innervation by sympathetic postganglionic neurons. The present study was thus aimed at determining the sympathetic postganglionic innervation of ICN. Choroids were processed for double immunofluorescence labelling with the following markers: tyrosine-hydroxylase (TH)/nitric oxide synthase (nNOS), TH/galanin (GAL), dopamine-beta-hydroxylase (DBH)/vasoactive intestinal polypeptide (VIP), TH/DBH and DBH/alpha-smooth-muscle actin (alphaSMA), and for triple immunofluorescence labelling with VIP/DBH/TH. Epifluorescence and confocal laser scanning microscopy were used for evaluation. Immunoperoxidase staining for TH or DBH in combination with NADPH-diaphorase histochemistry was applied for electron microscopy. ICN spread over the entire choroid but were concentrated in an equatorial zone passing obliquely from naso-cranial to temporocaudal. More than 80% of nNOS-positive ICN showed close appositions of TH/DBH-immunoreactive varicose nerve fibres at the light-microscopic level, as could be confirmed by confocal laser scanning microscopy. Ultrastructurally, these appositions could be defined as both synapses or close contacts without synaptic specialisation. Vascular and non-vascular smooth muscle fibres also received TH/DBH immunopositive innervation. Our findings suggest that most ICN receive a sympathetic input that might modulate their nitrergic effects upon vascular and non-vascular smooth muscle fibres in the choroid and that they may have more complex functions than merely being a simple parasympathetic relay. PMID- 11396713 TI - The spinal nerves innervate putative chemosensory cells in the ventral skin of desert toads, Bufo alvarius. AB - Toads normally obtain water by absorption across their skin from osmotically dilute sources. When hyperosmotic salt solutions are presented as a hydration source to dehydrated desert toads, they place the ventral skin onto the source but soon afterwards escape to avoid dehydration. The escape behavior coincides with neural excitation of the spinal nerves that innervate putative chemosensory cells in the ventral skin. In the present study, fluorescent dye translocated through the spinal nerves to those receptor cells in the epidermis was photoconverted in the presence of 3, 3'-diaminobenzidine tetrahydrochloride for electron-microscopic observation of the cells and associated nerve terminals. Most of the photoconverted cells were located in the deepest layer of the epidermis, with some being in more intermediate layers. No labeled cell was seen in the outermost layer of living cells. In desert toads, flask cells and Merkel cells are occasionally seen in the epidermis. An association of nerve fibers with these epidermal cells has been reported in some species of the anurans. In the present study, however, the cytological features of the photoconverted cells are neither reminiscent of flask cells nor Merkel cells, but are similar to those of surrounding epithelial cells in each layer of the epidermis. We hypothesize a sensory function for these cells, because they have a close association with nerve fibers and participate in the transepithelial transport of salts that must pass through all cell layers of the skin. PMID- 11396714 TI - Estrogen receptor-alpha and beta- immunoreactivity and mRNA in neurons of sensory and autonomic ganglia and spinal cord. AB - Estrogen receptor-alpha immunoreactivity and mRNAs are present in neurons in locales that innervate genital organs, e.g., parasympathetic pelvic autonomic ganglia, sensory dorsal root and nodose ganglia, and autonomic areas of the lumbosacral spinal cord. With the availability of probes for the beta-isoform of the estrogen receptor, we studied this receptor in autonomic, sensory, and spinal cord neurons and compared it with the distribution of the alpha-receptor. Estrogen receptor-alpha and -beta immunoreactivity were located in the nuclei of neurons, were in subpopulations of parasympathetic neurons in pelvic ganglia, and sensory neurons of dorsal root and nodose ganglia. Both receptor subtypes were present in the lumbosacral spinal cord: in neurons of the outer laminae of the dorsal horn, lateral collateral and medial collateral pathways, sacral parasympathetic nucleus, dorsal intermediate gray, and lamina X. Similar numbers of spinal cord neurons were immunoreactive for estrogen receptor-beta and estrogen receptor-alpha. However, estrogen receptor-beta-immunoreactive neurons appeared less numerous in the outer dorsal horn, but more numerous in the deeper layers of the spinal cord than estrogen receptor-alpha neurons. Retrograde tracing from the uterus revealed "uterine-related" neurons in dorsal root and pelvic ganglia that contained estrogen receptor-alpha and -beta. In situ hybridization revealed both estrogen receptor-alpha and -beta mRNA transcripts in sensory neurons of the dorsal root and nodose ganglia, parasympathetic neurons of pelvic ganglia, and spinal cord neurons in the dorsal horn, sacral parasympathetic nucleus, and dorsal intermediate gray of L6-S1 segments. These studies show that both estrogen receptor-alpha and -beta are synthesized by autonomic and sensory neurons in parts of the nervous system that have connections with the female reproductive system. Such neurons contain neurotransmitters that have important functions in the female reproductive organs; thus, it is likely that estrogen can influence the activity of such neurons and consequently, through them, the activities of the reproductive organs. PMID- 11396715 TI - Cellular localization of monoamine oxidase A and B in human tissues outside of the central nervous system. AB - We studied the localization of monoamine oxidase (MAO) A and B in human heart, liver, duodenum, blood vessels and kidney by immunohistochemistry. The primary antibodies used were mouse monoclonal anti-human MAO-A (6G11/E1) and anti-human MAO-B (3F12/G10/2E3). Samples were obtained from six routine autopsy cases and fixed in 2% paraformaldehyde. All cardiomyocytes and hepatocytes showed MAO-A and MAO-B immunoreactivity. In the duodenum, both immunoreactivities were present in all cells of the villi, Lieberkuhn crypts, muscularis mucosae and muscular layers, whereas Brunner glands were devoid of MAO-A and MAO-B staining. Endothelial cells of lymphatic vessels showed MAO-A but no MAO-B immunoreactivity, whereas arteries and veins presented MAO-A and MAO-B staining in muscular layers and fibroblasts but not in endothelial cells. In the kidney, renal tubuli showed MAO-A and MAO-B immunoreactivities, whereas collecting ducts and the Bowman's capsule showed only MAO-A staining. These data represent the first study of the cellular distribution of MAO-A and MAO-B in these human tissues. They show that both enzymes have a widespread distribution in the human body with a matching pattern in many, but not all tissues, and with strong differences from the pattern of distribution in rodents. PMID- 11396716 TI - Fluid phase endocytosis of [125I]iodixanol in rat liver parenchymal, endothelial and Kupffer cells. AB - Endocytosis of [125I]iodixanol was studied in vivo and in vitro in rat liver cells to determine fluid phase endocytic activity in different liver cells (hepatocytes, Kupffer cells and endothelial cells). The Kupffer cells were more active in the uptake of [l25I]iodixanol than parenchymal cells or endothelial cells. Inhibition of endocytic uptake via clathrin-coated pits (by potassium depletion and hypertonic medium) reduced uptake of [125I]iodixanol much more in Kupffer cells and endothelial cells than in hepatocytes. To gain further information about the importance of clathrin-mediated fluid phase endocytosis, the expression of proteins known to be components of the endocytic machinery was investigated. Using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting, endothelial cells and Kupffer cells were found to express approximately fourfold more rab4, rab5 and rab7 than parenchymal cells, while clathrin was expressed at a higher level in endothelial cells than in Kupffer cells and hepatocytes. Using electron microscopy it was shown that liver endothelial cells contained approximately twice as many coated pits per membrane unit than the parenchymal and Kupffer cells, thus confirming the immunoblotting results concerning clathrin expression. Electron microscopy on isolated liver cells following fluid phase uptake of horseradish peroxidase (HRP) showed that HRP-containing organelles had a different morphology in the different cell types: In the liver endothelial cells HRP was in small, tubular endosomes, while in Kupffer cells HRP was mainly found in larger structures, reminiscent of macropinosomes. Parenchymal cells contained HRP in small vacuolar endosomes with a punctuated distribution. In conclusion, we find that the Kupffer cells and the endothelial cells have a higher pinocytic activity than the hepatocytes. The hepatocytes do, however, account for most of the total hepatic uptake. The fluid phase endocytosis in liver endothelial cells depends mainly on clathrin-mediated endocytosis, while the parenchymal cells have additional clathrin-independent mechanisms that may play an important role in the uptake of plasma membrane components. In the Kupffer cells the major uptake of fluid phase markers seems to take place via a macropinocytic mechanism. PMID- 11396717 TI - Detection of P2X purinergic receptors on human B lymphocytes. AB - B lymphocytes are known to synthesise the P2X7 subtype of the P2X purinergic receptor family; however, the identification of the other six P2X subtypes on these cells has been limited by the absence of specific antibodies. In this study, we used a panel of anti-P2X polyclonal antibodies and confocal microscopy to examine the presence of each P2X receptor on human B lymphocytes. We observed that P2X1, P2X2, P2X4 and P2X7 subtypes, but not P2X3, P2X5 and P2X6 subtypes, are present on B lymphocytes. PMID- 11396718 TI - The distribution of tyrosine hydroxylase in the canary brain: demonstration of a specific and sexually dimorphic catecholaminergic innervation of the telencephalic song control nulcei. AB - Singing and the processing of auditory information related to song can be affected by experimental manipulations of catecholamine activity in the brain of zebra finches. We investigated, by immunocytochemistry in the brain of male and female canaries, the distribution of tyrosine hydroxylase (TH), the rate-limiting step in the synthesis of catecholamines. Fibers immunoreactive for TH (TH-ir) were particularly abundant in the lobus parolfactorius, the paleostriatum primitivum, and the nucleus septalis lateralis. A high density of TH-ir basket like structures was observed in the caudomedial neostriatum, an area involved in song perception and recognition. In most males, a high density of TH-ir fibers outlined the telencephalic song control nuclei including the high vocal center, the nucleus robustus archistriatalis, the nucleus interfascialis, the lateral and medial parts of the magnocellular nucleus of the anterior neostriatum, and area X of the lobus parolfactorius. The higher density of fibers immunoreactive for TH in these nuclei, compared with the surrounding telencephalon, supports the notion that the morphological evolution of the song control nuclei was accompanied by a neurochemical specialization. This specific innervation of the song control regions was, in general, not found in females. The specific presence of high densities of TH-ir fibers in the song system of male canaries and the sex difference of this innervation provide anatomical evidence in support of the claim that dopamine and/or norepinephrine play important roles in the modulation of song learning and production. PMID- 11396719 TI - Differential storage of hydroxyethyl starch (HES) in the skin: an immunoelectron microscopical long-term study. AB - Hydroxyethyl starch (HES) is widely used as a plasma substitute. Serious side effects occur only rarely, whereas a high incidence of severe pruritus has been reported. Moreover, tissue storage of HES has been demonstrated in various organs. The aim of the current study has been to examine precisely the intracellular uptake and long-term storage of HES in the skin. Skin biopsies from 119 patients who received HES of various preparations and cumulative dosage were obtained 30 min to 130 months after infusion therapy. The samples were analysed by ultrastructural and immunoelectron microscopy with HES-specific monoclonal and polyclonal antibodies. A characteristic vacuolisation of perivascular histiocytes was a regular finding in all skin biopsies as early as 1 day after a single infusion of 30 g. Immunoreactivity for HES was demonstrable within the vacuoles. Generally, the size and number of vacuoles in the histiocytes increased concomitantly with the cumulative dosage. Following administration of higher HES dosages, vacuoles were demonstrable in endothelial cells of blood and lymphatic vessels, basal keratinocytes, epithelia of sweat glands and in small peripheral nerves, the last mentioned being associated with pruritus. A subsequent reduction of the vacuoles in size and number could be demonstrated within 52 months. In nerves, HES deposits persisted no longer than 17 months paralleling the cessation of pruritus. Biopsies taken after 94 months exhibited no HES deposits in the skin. The condensation and final dissolution of the vacuoles may either indicate the release and subsequent redistribution of HES into the circulation or lysosomal degradation. PMID- 11396720 TI - Increased expression of non-muscle myosin heavy chain-B in connective tissue cells of hypertrophic rat urinary bladder. AB - Expression of the non-muscle myosin heavy chain-B (NM-MHC-B, also denoted as the embryonic smooth muscle myosin heavy chain, SMemb) was examined in rat urinary bladder during growth in response to a partial urinary outflow obstruction. Following obstruction, the weight of the urinary bladder increased more than five fold within 10 days. Immunohistochemistry with a polyclonal antiserum against the C-terminal sequence of NM-MHC-B revealed very few NM-MHC-B immunoreactive cells in the control urinary bladders. In hypertrophic bladders, the number of NM-MHC-B immunoreactive cells markedly increased. The majority of such cells were found in the interstitium surrounding smooth muscle bundles and also in the subserosal and submucosal layers. Western blot analysis showed that the NM-MHC-B expression was transient; the content of NM-MHC-B immunoreactive material had doubled 10 days after obstruction and then declined towards the control level after 6 weeks. Immunohistochemistry revealed co-localization of NM-MHC-B and vimentin within the same cells. NM-MHC-B did not co-localize with smooth muscle actin, suggesting that the source of NM-MHC-B is not a de-differentiated smooth muscle cell or myofibroblast but a non-muscle cell possibly reacting to tissue distension or stress. The NM-MHC-B-positive cells could have a role in the production of extracellular matrix and growth factors or be involved in modulation of spontaneous contractile activity. PMID- 11396721 TI - A rapid and strong apoptotic process is triggered by hyperosmotic stress in cultured rat cardiac myocytes. AB - In all cell types, the maintenance of normal cell volume is an essential homeostatic function. Relatively little is known about the induction of apoptosis by hyperosmotic stress and its molecular mechanism in terminally differentiated cardiac myocytes. We compared the apoptotic response of cultured neonatal rat cardiomyoctes to hyperosmotic stress by sorbitol (SOR) with those induced by doxorubicin (Doxo) or angiotensin II (Ang II). We also examined the apoptotic signaling pathway stimulated by the hyperosmotic stress. Apoptosis was assessed by the observation of: (1) cell viability, (2) DNA fragmentation detected by the TUNEL method and by agarose gel electrophoresis, and (3) poly(ADP ribose)polymerase (PARP) degradation, and Bcl-XS and Bcl-XL levels by Western blot analysis. Exposure of cardiomyocytes to 0.3 M SOR for 24 h resulted in decreased cell viability and increased generation of oligosomal DNA fragments (2.5-fold of controls). At this time, 83 +/- 5% of SOR-treated myocytes were TUNEL-positive (vs 23.7 +/- 6.8% in controls; P<0.01). PARP levels also decreased by approximately 42% when cardiac myocytes were exposed to SOR. Hyperosmotic stress induced a more rapid and stronger apoptotic response in cardiomyocytes than Doxo or Ang II. In addition, SOR increased 3.2-fold Bcl-XS proapoptotic protein without changes in Bcl-XL antiapoptotic protein levels and in the p53 transactivating activity. Taken together, these results strongly suggest that hyperosmotic stress triggers cardiac myocyte apoptosis in a p53-independent manner, being earlier and stronger than apoptosis induced by Doxo and Ang II. PMID- 11396722 TI - Absence of desmin slightly prolongs myoblast proliferation and delays fusion in vivo in regenerating grafts of skeletal muscle. AB - The expression of desmin, a muscle-specific intermediate filament protein, is upregulated during skeletal myogenesis, but its role in the myogenic process is unclear. Postnatal skeletal muscle regeneration occurs to completion in desmin null (-/-) mice, however, only late time points (i.e., days 7 and 21) in the myogenic process have been examined. This study observes the early events in skeletal muscle regeneration (i.e., from 3 days) in desmin (-/-) mice. Whole muscle autografts were performed in desmin (-/-) and control normal (Balb/c) mice. Muscle samples were taken on days 3, 5, 6, 7, 8, 9 and 11 after transplantation, and regeneration was assessed by graft morphology, patterns of cell proliferation and quantitation of myotube numbers. At day 5 myotube formation was delayed in the desmin (-/-) grafts compared to the normal controls. Immunohistochemical analysis of proliferating cell nuclear antigen demonstrated a very high proportion of proliferating cells in the periphery of desmin (-/-) whole muscle grafts at day 5 compared to the controls, where mitosis in this area was negligible. This strongly indicates t hat myoblast proliferation is prolonged during postnatal myogenesis in the absence of desmin. By day 6 there was no marked morphological difference between desmin (-/-) and normal control whole muscle grafts, although the zonal pattern of myoblast replication was slightly delayed in the desmin (-/-) mice until day 8. These results indicate a slightly extended phase of myoblast proliferation with delayed fusion in vivo in mature regenerating desmin (-/-) skeletal muscle. PMID- 11396723 TI - The embryonic development of the temnocephalid flatworms Craspedella pedum and Diceratocephala boschmai. AB - We have analyzed the embryonic development of the temnocephalid flatworms Craspedella pedum and Diceratocephala boschmai, using a combination of fuchsin labeled whole-mount preparation, histology, and transmission electron microscopy. Following the staging system recently introduced for another flatworm species (Mesostoma lingua), we can distinguish eight morphologically defined stages. Temnocephalids produce eggs of the neoophoran type in which a small oocyte is surrounded by a layer of yolk cells. Cleavage takes place in the center of the yolk mass (stages 1-2) and results in an irregular, multilayered disc of mesenchymal cells that moves to the future ventral egg pole (stage 3). Organ primordia, including those of the brain, pharynx, male genital apparatus, sucker, and epidermis "crystallize" within this disc without undergoing gastrulation movements (stage 4). An invagination of the epidermal primordium pushes the embryo back into the center of the yolk ("embryonic invagination"). As a result, organogenesis begins while the embryo is invaginated (stage 5). The brain differentiates into an outer cortex of cell bodies that surround a central neuropile. Precursor cells of the epidermis, pharynx, and protonephridia become organized into epithelia. During stage 6, the embryonic primordium everts back to the surface, where organogenesis and cell differentiation continues. Epidermal cells fuse into a syncytium that expands around the yolk. Myoblasts initially do not spread out in the way epidermal cells do; they remain concentrated in two narrow, longitudinal bands that extend along the sides of the embryo. Three pairs of axon tracts extending posteriorly from the brain follow the bands of myoblasts. Stages 7 and 8 are characterized by the appearance of eye pigmentation, brain condensation, and the formation of tentacles and a sucker that bud out from the epidermis of the anterior and posterior end, respectively. Comparison of morphogenesis in temnocephalids with observations in other flatworm taxa suggests a phylotypic stage for this phylum of invertebrates. PMID- 11396724 TI - Giant multinucleated macrophages occur in acute spinal cord injury. AB - Using a cell-isolation and -culture procedure specific for macrophages, we report the existence of giant (more than 50 microm diameter), multinucleated macrophages within an acute, 5-day-old adult rat spinal cord injury. The size and multinuclearity of these isolated giant cells was confirmed using transmission electron microscopy. Giant macrophages are markers for long-term infection, disease, and chronic injury in other soft tissues and are unexpected in the acute inflammatory stage of central nervous system injury. To our knowledge, this descriptive report is the first confirming the existence of giant macrophages in any injured nervous tissue, with additional data suggesting some of these cells to be multinucleated. PMID- 11396725 TI - Localization of an insulin-like peptide in brains of two flies. AB - A nucleotide sequence (GenBank AI134194) was identified in the database of the Berkeley Drosophila Genome Project based on the similarity of its translated sequence to insulin-like peptides known for other insects. The putative A chain of the Drosophila peptide was synthesized and used to produce an antiserum for immunocytochemistry. Medial neurosecretory cells and their axons were immunostained in whole brains of Drosophila larvae and adults. In larvae, immunostained axons from these cells extended to the corpus cardiacum in the ring gland and the hypocerebral ganglion and along the aorta in the head. In adults, axons from these cells extended along the aorta to the corpus cardiacum hypocerebral ganglion complex near the cardiac valve and branched along the midgut and crop. In contrast, two clusters of lateral neurosecretory cells and their axons were immunostained weakly in brains of female mosquitoes, Aedes aegypti. No other cells were stained in the nervous systems or midguts of Drosophila larvae and adults or Ae. aegypti females. These specific localizations substantiate the existence of an endogenous insulin-like peptide in Drosophila and suggest that a similar peptide is present in the distantly related mosquitoes. PMID- 11396726 TI - Uropathogenic Escherichia coli-induced neutrophil adhesion to urinary epithelium is strain-specific and mediated by CD11b/CD18. AB - Adherence of Escherichia coli to urinary tract epithelium induces neutrophil migration across the uroepithelium to combat bacterial infection. Neutrophil adherence to the apical membrane of uroepithelial cells may be an important factor for bacterial clearance. We used an in vitro model of urinary tract infection to examine the effects of uropathogenic E. coli on neutrophil adhesion to the uroepithelial cell line RT4. We found that distinct clinical isolates caused different levels of neutrophil adherence. One particular isolate caused significant neutrophil adhesion in a dose- and time-dependent manner. The neutrophil adhesion-promoting effect induced by this isolate was not caused by bacterial secreted products, suggesting that contact between intact E. coli and uroepithelial cells is required for promoting neutrophil adhesion. This adhesion was almost exclusively mediated by CD11b/CD18, suggesting that E. coli upregulates CD11b/CD18 counterligands on the uroepithelial surface. These data suggest that certain uropathogenic E. coli selectively promote adhesion of neutrophils to ligands on uroepithelial cells by a CD11b/CD18-dependent mechanism. PMID- 11396727 TI - The prevalence of chronic prostatitis-like symptoms in young men: a community based survey. AB - We surveyed the prevalence of chronic prostatitis-like symptoms in young men using the National Institutes of Health (NIH) Chronic Prostatitis Symptom Index (CPSI) and determined the clinical validity of the NIH-CPSI among men in the community. Of 29,017 men aged 20 years dwelling in the community, 8,705 men were randomly selected at a 30.0% sampling fraction and a total of 6,940 men (a response rate 79.7%) completed a self-administered questionnaire. Six percent reported having pain or discomfort in more than one area . About 5% did not feel that the bladder emptied fully after urinating more than 1 time in 5 and 10.5% had to urinate again within 2 h more than 1 time in 5. As the scores for pain or discomfort increased, those for urinary symptoms and impact on quality of life increased (P < 0.001; Armitage test). As the scores for urinary symptoms increased, those for pain or discomfort and impact on quality of life also increased (P < 0.001; Armitage test). The community-based prevalence of chronic prostatitis-like symptoms were found to be high in young men as well as in older men. Our findings indicate that men with pain or urinary symptoms experience a negative impact on their quality of life and the NIH-CPSI provides a valid measure for the general population. PMID- 11396728 TI - Biocompatibility of silver nitrate and ofloxacine coated bioabsorbable SR-PLLA rods. AB - The purpose of this study was to evaluate the biocompatibility of silver nitrate and ofloxacine coatings of bioresorbable self-reinforced poly-L-lactic acid (SR PLLA) rods. SR-PLLA rods coated with pure poly(caprolactone-co-L-lactide) or blended with silver nitrate (10, 5 or 2 weight-%) or ofloxacine (5 or 2 weight-%) were implanted in the dorsal muscles of 25 male rabbits. Tissue reactions caused by implantation trauma were seen 1 week after implantation. The positive control and 10 w-% silver nitrate coating showed the most marked reactions 1 month after implantation. Only sparse reactions were seen 6 months after implantation. Tissue reactions were scored semi-quantitatively. As a result of this study, we concluded that silver nitrate or ofloxacine coatings up to five w-% did not alter the good biocompatibility of SR-PLLA essentially. The method may lead to the possibility of preventing bacterial adhesion to urological stents during insertion. PMID- 11396729 TI - Clinical options for imipramine in the management of urinary incontinence. AB - Urinary incontinence is a common disorder in both childhood and adulthood. Proper treatment depends on insight into the pathophysiology and pharmacology of the lower urinary tract. This article reviews the mechanism of action of an old but commonly used drug, the tricyclic antidepressant agent imipramine, in nocturnal enuresis and stress and urge incontinence with reference to neuropharmacology and the relevant pathophysiology. PMID- 11396730 TI - Length dependence of the contractility of pig detrusor smooth muscle fibres. AB - Evidence on the length dependence of the contraction velocity of smooth muscle fibres is contradictory. Nevertheless, a thorough understanding of this dependence is essential for a correct urodynamic diagnosis of voiding problems. We studied muscle fibres of pig urinary bladders (n = 23). Force-velocity relations were measured at different muscle lengths with a stop test technique. This method involves measuring force generation of electrically stimulated muscle fibres during controlled shortening from a pre-shortening length at a pre-set velocity to a fixed stop-length. We normalized the length dependence of the measured properties to slack length, optimum length (the length at maximum isometric force generation), and passive force. Isometric force was found to be length dependent with an optimum length of 290+/-68% of the slack length (n = 11, P < 0.05). The maximum shortening velocity was 0.37+/-0.14 s(-1) related to the slack length and 0.13+/-0.05 s(-1) related to the optimum length and was not length dependent (n = 16, P < 0.05). Slack length is preferable to normalize the length dependence of smooth muscle. PMID- 11396731 TI - Alteration of neuronal and endothelial nitric oxide synthase and neuropeptide Y in congenital ureteropelvic junction obstruction. AB - PROBLEM: We investigated whether deranged nitric oxide synthase (NOS) and neuropeptide Y (NPY) expression is detectable in the stenotic segments of patients with congenital ureteropelvic junction obstruction. METHODS: Using real time reverse transcription-polymerase chain reaction (RT-PCR), we quantified mRNA amounts of NPY, neuronal (n), endothelial (e) and inducible (i) NOS in the stenotic segments of 20 patients with congenital ureteropelvic junction obstruction (aged 5.1+/-7.0 years) and of 21 unaffected controls (aged 23.5+/ 24.2 years). Additionally, mRNAs of glyceraldehyde-3-phosphate dehydrogenase (GAPDH), smooth muscle alpha-actin (Smactin), endothelial cell marker (CD31), and protein gene product 9.5 (PGP 9.5) were evaluated. Immunohistochemistry was made for NPY, nNOS, eNOS, iNOS, PGP 9.5, and CD 31. RESULTS: The mRNA of nNOS was significantly reduced in the obstructed junctions when related to the mRNAs of Smactin (P < 0.001) or GAPDH (P < 0.05), respectively. A significant reduction was also obtained for eNOS mRNA when standardized to CD31 (P < 0.05), GAPDH or Smactin mRNA (P < 0.05, and P < 0.001, respectively). NPY, PGP 9.5 and iNOS mRNAs were found in comparable quantities in both groups. In the stenotic segments, Smactin mRNA level was about twofold higher than in our control specimens, as shown by the lower CT values for the patients in real-time PCR (16.9+/-2.0 vs 17.9+/-2.6, P < 0.05). Furthermore, Smactin, nNOS, iNOS, eNOS, and NPY mRNA levels in specimens of unaffected ureteropelvic junctions were independent of age. Major differences between control and stenotic tissues were detected by immunohistochemistry: There was a dramatic reduction of innervation density as evidenced by nNOS and NPY labeling. CONCLUSION: Taken together, we found alterations in NOS gene expression and NPY innervation in tissue specimens of patients with congenital ureteropelvic junction obstruction. PMID- 11396732 TI - Crystallisation properties in stone forming and normal subjects' urine diluted using a standardised procedure to match the composition of urine in the distal part of the distal tubule and the middle part of the collecting duct. AB - Using a standardised procedure, we assessed the crystallisation properties of calcium phosphate in urine with a composition matching that in the distal part of the distal tubules (DTd) and of calcium oxalate in urine with a composition matching that in the mid-collecting duct (CDm). We used 8-h urine samples collected between 2200 h and 0600 h with sodium azide as preservative. Urine from ten patients with recurrent CaOx stone formation and from ten normal subjects was used for the measurements. The DTd and CDm samples were obtained by diluting the voided 8-h urine to 3000 ml and 1750 ml per 1.73 m2 body surface area, respectively. The nucleation was studied in DTd urine following supersaturation with CaP. The crystal size distribution was assessed with a Coulter counter both following supersaturation of DTd urine with CaP and of CDm urine with CaOx. The crystallisation of CaP in DTd urine as well as that of CaOx in CDm urine, in the presence of CaP crystals that had been precipitated in DTd urine, was measured with the isotope technique. The inhibition of CaOx and brushite crystal aggregation in standardised diluted aliquots of DTd and CDm urine was assessed spectrophotometrically as the rate of sedimentation. There was a slightly increased sedimentation rate and a lower initial absorbance in DTd urine from stone formers supersaturated with CaP. Although these findings might reflect a state of increased crystal aggregation in stone formers' urine, this could not be confirmed by crystal size measurements in the Coulter counter. The inhibition of brushite crystal aggregation in DTd urine was significantly in stone formers' urine than in normal subjects' urine (P < 0.001). Moreover, all inhibition values in DTd samples from stone formers were negative, suggesting a promoter effect on crystal aggregation. The inhibition of CaOx crystal aggregation in CDm urine also was significantly higher in CDm urine from normal subjects than in CDm urine from stone formers (P < 0.05). For all other variables the level was similar when urine samples from the two groups were compared. Although this series of crystallisation assessments was carried out on a small number of standardised diluted urine samples only, the results nevertheless emphasise a defect in aggregation inhibition as one important determinant for an abnormal calcium salt crystallisation in patients with recurrent stone formations. This difference obviously includes aggregation of both CaP crystals in DTd urine and CaOx crystals in CDm urine. The results also show that assessment of crystallisation properties of this kind can be carried out in standardised, diluted 8-h night urine samples, which accordingly can be used in the routine work-up of patients with calcium stone disease. Such an approach might prove useful in order to get information on the combined effects of the driving force of supersaturation and crystallisation modifying properties accomplished by urinary macromolecules and other modifying agents. PMID- 11396733 TI - A study of crystal matrix extract and urinary prothrombin fragment 1 from a stone prone and stone-free population. AB - South African blacks are immune to urinary calculi whereas whites have an incidence rate similar to that reported in Western societies. Urinary prothrombin fragment 1 (UPTF1) and the crystal matrix extract (CME) from which it is derived have been shown to be potent inhibitors of crystal growth and aggregation in undiluted human urine. The objective of the present study was to isolate CME and UPTF1 from the urines of black and white subjects in order to assess whether either might contribute to the black population's relative stone immunity. CME was isolated from freshly precipitated calcium oxalate (CaOx) crystals and a crystallization study was conducted in synthetic urine. Coulter Counter, 14C oxalate deposition, and scanning electron microscopy data demonstrated that the extracts from both race groups strongly inhibited CaOx nucleation. The extract derived from the black subjects inhibited nucleation to a greater extent than that from the whites. A phase conversion from COM to COD in the presence of the extracts, in support of the inhibitory effect of CME, was also observed. Purified UPTF1 isolated from both groups' CME was subjected to rigorous biochemical characterization involving matrix-assisted laser desorption ionization time-of flight (MALDI-TOF) mass spectrometry, protein sequencing by Edman degradation, and amino acid analyses. No differences in molecular weight or amino acid sequence and composition were identified. It is suggested that the more potent inhibitory activity of the extract derived from the black subjects might be related to this group's relative stone immunity. PMID- 11396734 TI - The reappraisal of nephrocalcin--its role in the inhibition of calcium oxalate crystal growth and interaction with divalent metal ions. AB - Nephrocalcin (NC) is a potent crystal growth inhibitor of calcium oxalate monohydrate. However, the sequence is undefined owing to its multiple glycosylations. Although there have been many biochemical studies of the binding capacity of calcium, the study of the function of the domain is still deferred. By using S-200 gel filtration and Mono Q ion-exchange chromatographic procedures, NC can be purified without further treatment for the removal of urobilirubin. The kinetic study of crystal growth in calcium oxalate monohydrate is performed using a seed slurry system. NC was cut into two peptides through interaction with copper ion and ascorbic acid. The interaction site of the copper ion is presumed to be located between 8 and 6 kDa of molecular weight in NC. The data suggest that divalent metal ions may be involved in the calcium oxalate crystallization through interaction with NC. The role of ascorbic acid in the formation of urinary stones should be reappraised for its association in the redox reaction, with resultant protein digestion in the presence of copper ions. PMID- 11396735 TI - Arginine form of p21 gene codon 31 is less prominent in patients with calcium oxalate stone. AB - The formation of urinary stones is associated with cell death in response to various injuries. P21 (WAF1/CIP1) is a downstream protein of P53 and can arrest the cell cycle at G1/S with resulting cell death. We aimed to investigate the polymorphism of p2 gene codon 31 as the genetic marker in searching for the association of urolithiasis. One hundred and nineteen healthy controls and 95 patients with calcium oxalate stone were examined in this study. The polymorphism was seen from the result of polymerase chain reaction-based restriction analysis. The result revealed significant differences between normal individuals and stone patients (P < 0.05) and the distribution of arginine homozygote in the control group (31.9%) was higher than in the patient group (16.8%). It is concluded that polymorphisms of p21 codon 31 can be a genetic marker for urinary stone disease. Individuals possessing arginine form of p21 codon 31 have less risk of developing calcium stone disease. PMID- 11396736 TI - Osteocalcin gene Hind III polymorphism is not correlated with calcium oxalate stone disease. AB - The formation of urinary stones is presumed to be associated with polymorphism of the osteocalcin gene. The most frequently seen polymorphism is the Hind III type located at the promoter region. This polymorphism has been used as a genetic marker in the search for a correlation between urolithiasis and normal subjects. In our study, a normal control group of 105 healthy people and 102 patients with calcium oxalate stones were examined. The polymorphism was seen following polymerase chain reaction-based restriction analysis. The results revealed no significant differences between normal individuals and stone patients (P = 0.978), and distribution of the TT homozygote in the control group (42.9%) was similar to that in the patient group (42.2%). Further categorization of the stone patients into normocalciuric and hypercalciuric groups also revealed no statistical differences from controls. We conclude that Hind III polymorphism of the osteocalcin gene is not a suitable genetic marker of urinary stone disease. Further searches for other polymorphisms on this gene correlated with stone disease are suggested. PMID- 11396737 TI - Proton magnetic resonance spectroscopy in brain tumours: clinical applications. AB - Parallel to the rapid development of clinical MRI, MR spectroscopy (MRS) has, after starting as an analytical tool used in chemistry and physics, evolved to a noninvasive clinical examination. Most common neuroradiological diagnostic indications for MRS are functional inborn errors, neonatal hypoxia, ischaemia, metabolic diseases, white matter and degenerative diseases, epilepsy, inflammation, infections and intracranial neoplasm. Compared to CT and MRI, well established morphological diagnostic tools, MRS provides information on the metabolic state of brain tissue. We review the clinical impact of MRS in diagnosis of tumours and their differentiation from non-neoplastic lesions. PMID- 11396738 TI - Diagnostic potential of short echo time MR spectroscopy of gliomas with single voxel and point-resolved spatially localised proton spectroscopy of brain. AB - Accurate neuroimaging grading of gliomas is useful for management, but techniques such as MRI and CT are not sufficiently reliable. Necrosis is a consistent, decisive prognostic factor and the key diagnostic criterion for glioblastoma multiforme. MR spectroscopy (MRS) allows noninvasive measurement of metabolites in brain tumours and mobile lipids reflect necrosis. However, short echo-time (TE) spectroscopy has been required for reliable assessment of lipids, since their relaxation times are very short. Recent advances have made it possible to perform short-TE MRS. We attempted to evaluate the significance of short TE spectroscopy as part of routine imaging for diagnosis and grading of gliomas. We performed TE 30 ms MRS in 25 patients with gliomas (grade II six; grade III three; grade IV, 16) and in 19 areas of healthy white matter using proton brain examination/single voxel (PROBE/SV) and point-resolved spatially localised spectroscopy (PRESS). With short-TE spectroscopy, lipid signals were detected in all 16 tumours of grade IV, one grade II (P = 0.0002) and none of grade III (P = 0.001). TE 136 ms MRS, carried out in 20 of these cases, showed lipid signals in only four of 14 grade IV tumours and in none of the other six. N acetylaspartate/choline (NAA/Cho) ratios were always more than 1.0 in healthy tissues and less than 1.0 in all but one of the gliomas. The mean creatine (Cr)/Cho ratio in each tumour grade was significantly lower than in the healthy tissues. The mean Cr/Cho ratio was also significantly lower in grade IV than in grade II tumours (P < .0005). Considerable overlap in Cr/Cho ratio was observed between grade II and grades III and IV gliomas at long but less so at short-TE MRS. We conclude that short-TE MRS with PROBE/SV and PRESS is of value in grading gliomas. PMID- 11396739 TI - High-resolution blood oxygen-level dependent MR venography (HRBV): a new technique. AB - Our purpose of this study was to demonstrate the clinical potential and spatial resolution of a new MRI technique: high-resolution blood oxygen-level dependent venography (HRBV), in well-known intracranial vascular lesions, such as cavernous and venous angiomas, and venous sinus thrombosis. HRBV provides unique high resolution information on veins without administration of contrast medium. The data are independent of conventional findings on MRI and potentially useful in characterising and demonstrating the architecture of vascular lesions of the brain. PMID- 11396740 TI - Cerebral abscesses: investigation using apparent diffusion coefficient maps. AB - The combination of high signal and reduced apparent diffusion coefficients (ADC) within abscesses on diffusion-weighted MRI (DWI) has been reported as characteristic of abscesses, and useful for distinguishing them from cystic or necrotic neoplasms. To assess whether these are consistent findings in abscesses, we used DWI-derived ADC to investigate changes in water diffusibility in cerebral abscesses. We reviewed the MRI studies and clinical records of five patients with brain abscesses, who underwent DWI. Regions of interest were drawn within the abscesses on ADC maps, to obtain the ADC. The center of all five abscesses gave signal higher than that of white matter on DWI. The three largest also appeared bright on ADC maps, i. e., showed ADC substantially lower than those of normal white matter, consistent with restricted diffusion. However, the two smaller abscesses were not visible on ADC maps because their ADC were essentially the same as that of white matter; they did not show restricted diffusion. The absence of restricted diffusion within small abscesses may be related to intrinsic differences in molecular microenvironment between small and large abscesses, or to greater influence of volume averaging with surrounding edema on the ADC in smaller abscesses. PMID- 11396741 TI - A ruptured dissecting aneurysm of the vertebral artery: comparison of angiographic and histological findings. AB - We present autopsy findings in a patient with a dissecting aneurysm of the vertebral artery causing subarachnoid haemorrhage. We sectioned the artery longitudinally and compared it with the angiogram, which showed the "pearl-and string". Histological examination showed a pseudoaneurysm covered by adventitia alone, forming the "pearl". Internal elastic lamina and media were destroyed, and haematoma extended in the subadventitial space of the wall of the pseudoaneurysm. Media thickened by haematoma caused the "string", narrowing the parent artery. Alcian blue staining showed that stratified internal elastic lamina in the aneurysm and the parent artery, had undergone marked mucoid degeneration, which may have been responsible for the dissection. PMID- 11396742 TI - The effects of cidofovir on progressive multifocal leukoencephalopathy: an MRI case study. AB - MRI was used to study the effects of introducing cidofovir (HPMPC, Vistide) to the antiretroviral therapy of a 33-year-old white man diagnosed as having progressive multifocal leukoencephalopathy (PML) secondary to AIDS. In response to combined cidofovir and antiretroviral therapy he showed significant clinical improvement. MRI showed a decrease in extent of existing lesions, without new ones. Blood chemistry information obtained indicated some involvement of immunologic mechanisms: the CD4:8 ratio showed improvement from an average of 0.08 before treatment to 0.13 during therapy. PMID- 11396743 TI - The natural history of transdural herniation of the spinal cord: case report. AB - We report a patient with a Brown-Sequard syndrome who developed ventral transdural spinal cord herniation, showing the imaging findings changing over time from two thoracic disc protrusions with a normal spinal cord to a cord trapped within a bony defect in a thoracic vertebra. PMID- 11396744 TI - High-intensity facial nerve lesions on T2-weighted images in chronic persistent facial nerve palsy. AB - Our aim was to estimate the value of MRI in detecting irreversibly paralysed facial nerves. We examined 95 consecutive patients with a facial nerve palsy (14 with a persistent palsy, and 81 with good recovery), using a 1.0 T unit, with T2 weighted and contrast-enhanced T1-weighted images. The geniculate ganglion and tympanic segment had gave high signal on T2-weighted images in the chronic stage of persistent palsy, but not in acute palsy. The enhancement pattern of the facial nerve in the chronic persistent facial nerve palsy is similar to that in the acute palsy with good recovery. These findings suggest that T2-weighted MRI can be used to show severely damaged facial nerves. PMID- 11396745 TI - The two-in-one model: a new variation of the arteriovenous malformation model in swine. AB - We report a new variation of the well-established experimental arteriovenous malformation (AVM) model in swine. To provide high flow through the rete mirabile (nidus, RM) and thereby to reduce the rate of spontaneous thrombosis of the AVM, we performed an end-to-end anastomosis of the left common carotid artery (CCA) and the external jugular vein (EJV) microsurgically in three micropigs. After 1 and 4 months the animals underwent angiograms of the CCA and vertebral artery (VA). In all cases the diversion of the blood through the RM was patent, up to the 4 months follow-up. We observed an arteriovenous fistula (arteriovenous pseudomalformation, pAVF) between the VA and the EJV in each case at both 1 and 4 months. This modification of the well-known AVM model in the micropig could be used to monitor long-term changes after embolisation, avoiding the naturally high rate of spontaneous thrombosis. This two-in-one model is thus well suited for preclinical testing of embolic materials. PMID- 11396746 TI - Fatal rebleeding following coil embolization of cerebral aneurysms: the role of long-term systemic anticoagulation. AB - Embolization of cerebral aneurysms has become a common technique. Its impact on subsequent medical management of the patient is not well known. We report two patients who presented in a poor neurological grade after subarachnoid hemorrhage from posterior communicating artery aneurysms. Both were treated by coil embolization and both developed subclavian vein thrombosis, requiring systemic anticoagulation, initiated 11 and 21 days after embolization, respectively. Both developed a large, fatal intracranial hemorrhage adjacent to the embolized aneurysm in the fourth week of anticoagulation. Systemic anticoagulation of patients who have had a ruptured aneurysm treated by coil embolization may carry a significant risk of rebleeding. Alternate management strategies should be considered in these patients. PMID- 11396747 TI - Posterior cranial fossa arteriovenous fistula with presenting as caroticocavernous fistula. AB - We report cases of posterior cranial fossa arteriovenous fistula (AVF) with presenting with exophthalmos, chemosis and tinnitus in 26- and 66-year-old men. The final diagnoses was vertebral artery AVF and AVF of the marginal sinus, respectively. The dominant venous drainage was the cause of the unusual presentation: both drained from the jugular bulb or marginal sinus, via the inferior petrosal and cavernous sinuses and superior ophthalmic vein. We used endovascular techniques, with coils and liquid adhesives to occlude the fistulae, with resolution of the symptoms and signs. PMID- 11396748 TI - Percutaneous coaxial transpedicular biopsy of vertebral body lesions during vertebroplasty. AB - We evaluated the safety and histological results of percutaneous transpedicular biopsy in patients undergoing vertebroplasty for vertebral collapse. Over a 6 year period, we carried out biopsies in 46 patients who underwent percutaneous injection of acrylic surgical cement for 57 collapsed vertebrae, because the diagnosis was not clearly established on clinical or imaging grounds. All procedures were performed under fluoroscopic guidance via a coaxial bitranspedicular approach used for vertebroplasty. We performed a clinical examination and CT after every procedure and approximately 6 months thereafter. Biopsies contributed to in 55 (96.5%) of the 57 vertebral lesions. Biopsy material was inadequate in one case (1.7%) and one biopsy was a false-negative (1.7%). The accuracy of the histological results was 98.2%, allowing a correct diagnosis in 55 of the 56 procedures. Of the 37 lesions in 28 patients with a history of a tumour, the final diagnosis was osteoporotic collapse in 25 (67.6%), metastasis in nine (24.3%), and myeloma in three (8.1%). The final diagnosis in the 19 lesions in 17 patients without a known tumour was osteoporotic collapse in 12 (63.2%), metastasis in five (26.3%), and amyloidosis in two (10.5%), the latter in one patient. No complications were observed. PMID- 11396749 TI - Emedastine-ketoconazole: pharmacokinetic and pharmacodynamic interactions in healthy volunteers. AB - OBJECTIVE: To assess the pharmacokinetic and pharmacodynamic interactions of emedastine difumarate, a new antihistamine drug and ketoconazole. MATERIAL: Twelve healthy Caucasian volunteers were administered emedastine difumarate 4 mg oral capsules once daily for 10 consecutive days. From day 6 to day 10, ketoconazole 200 mg were co-administered twice daily. METHODS: The effects of multiple ketoconazole administration on emedastine kinetics were evaluated by comparing values obtained for pharmacokinetic parameters at steady state, with and without ketoconazole. C(ss,max), C(ss,min), tmax, AUCss, t(1/2) and Cl(ss)/F values, obtained after both treatments, were compared. Significant difference was defined as p < 0.05. QTc intervals from ECGs at baseline, after emedastine treatment and after emedastine-ketoconazole co-treatment were statistically compared. RESULTS: Emedastine steady state pharmacokinetics were slightly altered as a result of the ketoconazole co-treatment. AUCss rose by about 33% (increase ranging from 0.96 to 66.86, p < 0.001) and total clearance decreased by about 30% (ranging from 0.96 to 40.08, p < 0.001) with no change in the half-life. These events did not lead to relevant pharmacodynamic changes, i.e. maximum prolongation of the corrected QT interval (QTc) observed after 5 days co treatment (day 10) was of about 4%. Rate and severity of anti-H 1 sedation episodes also did not increase on ketoconazole co-treatment. CONCLUSIONS: A moderate, but statistically significant interaction between emedastine and ketoconazole was observed. Pharmacodynamic data indicate no increase in the QTc interval during concomitant therapy. This result is consistent with the multiple emedastine metabolic pathways shown in man which supplement the metabolism by different enzymatic isoforms of CYP450. Concomitant treatment with emedastine and ketoconazole in subjects with normal QT intervals can therefore, be undertaken without special precautions. PMID- 11396750 TI - A randomized trial to compare the efficacy, safety, cost and platelet aggregation effects of enoxaparin and unfractionated heparin (the ESCAPEU trial). AB - OBJECTIVE: To compare the efficacy, safety, cost and effects on platelet aggregation of unfractionated heparin and low-molecular weight heparin in unstable angina patients. PATIENTS AND METHODS: Ninety-three patients with unstable angina were randomized to receive either unfractionated heparin (UFH) or enoxaparin in an open design clinical trial with blinded end point evaluation. The effects of the heparins on platelet aggregation were also compared. RESULTS: The composite end point of myocardial infarction, cardiac death, recurrent angina and need for intervention was observed in 62% of patients treated with UFH and in 37% of patients treated with enoxaparin (RR 1.7, 95% CI 0.75 to 3.71, p = 0.04). There was no difference in the frequency or severity of adverse events. A cost effectiveness analysis showed both the heparins to be similar. Platelet aggregation was inhibited to a greater extent by UFH when compared to enoxaparin. CONCLUSIONS: Enoxaparin appears to be superior in efficacy to UFH and similar to UFH in safety. No difference in costs was detected in this study. The greater inhibition of platelet aggregation observed in the case of UFH compared to enoxaparin indicates that there may be more bleeding complications with UFH. PMID- 11396751 TI - A comparative study of caudal bupivacaine and midazolam-bupivacaine mixture for post-operative analgesia in children undergoing genitourinary surgery. AB - OBJECTIVE: This study was designed to evaluate the analgesic efficacy of caudal midazolam-bupivacaine combination in providing post-operative pain relief in children undergoing genitourinary surgery and to study the occurrence of adverse effects. SUBJECTS AND METHODS: Thirty children, aged 2 to 8 years, scheduled for genitourinary surgery were allocated randomly to receive either 0.25% bupivacaine 0.5 ml/kg (group B; n = 15) or 0.25% bupivacaine 0.5 ml/kg with 50 microg/kg midazolam (group BM; n = 15) by the caudal route immediately after induction of general anesthesia. Heart rate, arterial blood pressure and oxygen saturation were monitored throughout the study period. Postoperative pain was assessed at regular intervals for 12 hours using an objective pain score. Analgesia was supplemented whenever the pain score was > or = 4. Duration of analgesia, as well as the requirement of additional analgesics, were noted. RESULTS: Lowest pain scores were observed with the addition of midazolam to caudal bupivacaine (p < 0.01). Duration of analgesia was longer in group BM (11 +/- 0.5 h) as compared to group B (7.4 +/- 2.1 hours) (p < 0.05). Fewer children (26.6%) required additional analgesia in the combination group whereas in group B, 60% of the children received analgesic supplements within 6 hours after surgery (p < 0.05). There were no significant changes in heart rate, blood pressure and oxygen saturation in both groups. We observed no untoward event in either of the groups. CONCLUSION: Caudal administration of bupivacaine-midazolam mixture prolongs post operative analgesia compared to bupivacaine alone without causing any adverse effects. PMID- 11396752 TI - Protective effects of fosfomycin on cisplatin-induced nephrotoxicity in patients with lung cancer. AB - SUBJECTS, MATERIAL AND METHODS: Protective effects of fosfomycin on cisplatin induced nephrotoxicity have been previously reported, however, the proper time, duration and dosage of its administration were uncertain. Therefore, we investigated the protective effect of concurrent administration of twice-daily doses of 2 g fosfomycin for 5 days in 13 cisplatin-naive lung cancer patients who were due to receive a single dose per cycle of 100 mg/m2 cisplatin. On each chemotherapeutic cycle, patients were randomly given cisplatin alone or cisplatin plus fosfomycin every 4 weeks for a maximum of 4 consecutive cycles. Indicators of nephrotoxicity, urinary N-acetyl-beta-D-glucosaminidase (NAG) activity, serum creatinine (Scr) and creatinine clearance (Clcr) were determined the day before and at day 3 and day 6 after cisplatin administration. Results were compared and statistically analyzed by the non-parametric Mann-Whitney's test. We found that the NAG activities obtained on day 0, day 3 and day 6 of the fosfomycin cycles were comparable to values obtained during the control cycles (p > 0.05). Moreover, the NAG activities on day 3 of both treatment cycles were significantly elevated from baseline (p < 0.01) and had normalized on day 6. There were no significant changes in serum creatinine and creatinine clearance. CONCLUSION: High-dose cisplatin induced reversible elevation of urinary NAG and concurrent administration of low-dose fosfomycin for 5 days had no effect on the enzymuria. In the prevention of cisplatin nephrotoxicity, a further study using dose escalation (8 to 12 g/d) of fosfomycin administered 2 to 3 days prior to cisplatin are required to demonstrate its nephroprotective effects. PMID- 11396753 TI - Effect of age and gender on the pharmacokinetics of ebastine after single and repeated dosing in healthy subjects. AB - OBJECTIVES: Ebastine is a potent and selective H1-receptor antagonist indicated for allergic rhinitis which undergoes extensive first pass metabolism by CYP3A4 to form an active metabolite, carebastine. The purpose of the study was to determine age- and gender-related differences in the pharmacokinetics of ebastine and carebastine. METHODS: The upper recommended oral dose of 20 mg once daily was administered to 12 healthy young (22 to 38 years) and 12 healthy elderly (50 to 92 years; 8 m and 4 f) subjects for 5 days. Plasma concentrations of ebastine and carebastine were determined for 24 hours following the initial dose on Day 1 and for 72 hours following the dose on Day 5 using a sensitive LC/MS/MS assay. The minimum quantifiable limit (MQL) for the assay was 0.05 ng/ml and 1.0 ng/ml for ebastine and carebastine, respectively. RESULTS: Mean area under the curve and Cmax values on Day 1 and Day 5 were similar for ebastine but approximately doubled for carebastine due to its longer half-life. Mean carebastine concentrations were approximately 10 to 20 fold higher than mean ebastine concentrations. For young subjects, the mean (%CV) ebastine t(1/2) was 5.76 (28.47) h and 20.38 (46.19) h on Day 1 and Day 5, respectively. Similarily, for young subjects, the mean (%CV) for carebastine t(1/2) was 7.03 (23.21) h and 26.12 (23.39) h on Day 1 and Day 5, respectively. This apparent prolongation of t(1/2) was probably due to lack of proper estimation of terminal half-life on Day 1 as fewer samples were collected for a shorter duration on Day 1. Using a multicomparison test for Cmin values, it was determined that steady state conditions were achieved by Day 5 for both age groups for ebastine and in young subjects for carebastine. The variability in ebastine pharmacokinetic parameters was higher than carebastine. A 50% increase in ebastine AUC(0-24) and Cmax values in elderly subjects, with no changes in t(1/2), could be explained by either increased absorption of ebastine in elderly subjects or due to a decrease in first pass metabolism. As ebastine shows a high first-pass effect, even a small change in this first pass can cause large changes in plasma exposure. The ebastine pharmacokinetic parameters for elderly subjects in this study lie between the values reported in young subjects in earlier studies. Hence, the apparent age-related pharmacokinetic difference for ebastine is probably due to the inherent variability in ebastine pharmacokinetics. There were no gender related differences in either young or elderly subjects for mean AUC, Cmax, tmax and t(1/2) ebastine and carebastine values. Ebastine was absorbed rapidly with a median tmax of 1.25 to 2.25 h for both healthy young and elderly males and females on Day 1 and Day 5. There was a delayed appearance of carebastine as expressed by median tmax of 4.0 to 5.0 h, which did not change with age, gender or repeated administration. There were no clinically relevant differences between the groups of subjects with respect to adverse events or safety parameters. CONCLUSIONS: Thus, ebastine can be safely administered to elderly subjects with no clinically important age- or gender related differences in the pharmacokinetics of ebastine/carebastine. PMID- 11396754 TI - A comprehensive account on the role of efflux transporters in the gastrointestinal absorption of 13 commonly used substrate drugs in humans. AB - BACKGROUND: The potential absorption-limiting effect of intestinal efflux transporters such as P-glycoprotein (P-gp) has been well recognized, primarily based on results of numerous Caco-2 cell studies showing that flux, permeability, or transport clearance of drugs from the basolateral to the apical (B --> A) compartment is greater than that from the apical to the basolateral (A --> B) compartment. Except for very limited examples such as celiprolol, talinolol, pafenolol and paclitaxel, the potential clinical impact of these transporters on oral absorption of the vast number of commonly prescribed drug substrates in humans has not been closely examined to date. OBJECTIVE: To evaluate whether these efflux transporters may play a significant role in limiting oral absorption of 13 commonly used drugs (digoxin, etoposide, felodipine, fexofenadine, furosemide, indinavir, losartan, nadolol, propranolol, ritonavir, saquinavir, tacrolimus, and verapamil) in humans. METHODS: Drug absorption properties such as the rate (as judged by the Cmax and tmax) and extent (as judged by AUC or urinary excretion of drugs) of absorption as a function of dose, as well as the completeness of oral absorption were obtained from the literature. RESULTS: The absorption properties of these 13 drugs are not consistent with absorption retarding expectations from in vitro studies because they all show apparent dose independent kinetics in absorption or bioavailability and completeness of oral absorption is shown for most of the drugs evaluated. CONCLUSIONS: In spite of being substrates of intestinal efflux transporters such as P-gp, the in vivo oral absorption of 13 drugs examined apparently is not significantly impeded by efflux transporters. Thus, there may exist an apparent discrepancy between in vitro "expectations" and in vivo results; potential reasons for this are discussed. The present findings, however, do not de-emphasize potential in vivo importance of efflux transporters in affecting (increasing or decreasing) oral absorption of certain substrate drugs, especially those with low to moderate intestinal permeability and with low therapeutic index, or the importance of efflux transporters in the study of mechanisms of drug absorption and some potentially clinically significant drug-drug and drug-food interactions. PMID- 11396755 TI - Methylene blue staining of dysplastic and nondysplastic Barrett's esophagus: an in vivo and ex vivo study. AB - BACKGROUND AND STUDY AIMS: Methylene blue selectively stains specialized columnar epithelium in Barrett's esophagus with high accuracy. We prospectively evaluated the methylene blue staining properties of dysplastic and nondysplastic Barrett's esophagus and the association of these properties with the risk for dysplasia and cancer. PATIENTS AND METHODS: In a ex vivo study, we mapped, photographed, and sampled esophagectomy specimens with high grade dysplasia and/or early adenocarcinoma before and after methylene blue staining. In a concurrent in vivo study, we performed methylene blue staining and characterized methylene blue stain characteristics. Pathologists estimated the proportion of specialized columnar epithelium in each specimen and graded dysplasia. RESULTS: We examined 551 biopsies from 47 patients with biopsy-proven Barrett's esophagus and 48 sections from five surgical specimens with Barrett's esophagus and dysplasia and early adenocarcinoma. The accuracy of ex vivo and in vivo methylene blue staining for specialized columnar epithelium was 87% and 90%, respectively. It was influenced by the length of Barrett's esophagus, biopsy location, and the presence of esophagitis and/or dysplasia. Light to absent staining (p = 0.01) and moderate to marked heterogeneity (p = 0.01) were significantly associated with high grade dysplasia or cancer in the univariate analysis and in a multivariate model that adjusted for the length of Barrett's esophagus and the presence of a lesion. These staining characteristics were present in all patients with severe dysplasia and/or adenocarcinoma. CONCLUSIONS: Highly dysplastic or malignant Barrett's esophagus stains differently with methylene blue. Increased heterogeneity and decreased methylene blue stain intensity are significant independent predictors of high grade dysplasia and/or cancer. These features may help to direct biopsies in patients without a lesion. PMID- 11396756 TI - In vitro study and in vivo application of a reusable double-channel sphincterotome. AB - BACKGROUND AND STUDY AIMS: Therapeutic endoscopic retrograde cholangiopancreatography (ERCP) has been deemed to be a "cost-prohibitive" procedure, based upon the cumulative costs of one-time-use accessories and current reimbursement plans. One-time-use sphincterotomes comprise a significant component of that cost and, accordingly, we evaluated the disability and clinical usefulness of a recently introduced reusable double-channel sphincterotome. MATERIALS AND METHODS: We studied a reusable 6-Fr sphincterotome at baseline and following contamination with 10(6) Bacillus stearothermophilus. Reprocessing included a unique 30-minute ultrasonic cleaning step in lieu of manual cleaning, followed by steam sterilization. Parameters evaluated included sphincterotome function, electrical integrity, and our ability to sterilize the devices for three in vitro trials. In vivo studies included patient demographics and outcomes, procedural findings, and success rates, and the mean number of times the sphincterotome was used, functional grading at time of use, and reasons for sphincterotome malfunction. RESULTS: Ten out of ten sphincterotomes maintained form, function, and electrical integrity in vitro, and all cultures were negative after sterilization. In the initial in vivo study, ten sphincterotomes were used in 50 patients (mean, 5 uses) with a 94% success rate. Reasons for sphincterotome failure included leak or breakage of the accessory port in 70%, wire fracture in 10%, incorrect wire bow in 10%, and clogged injection port in 10%. Following reconfiguration of the insertion-port polymer, an additional ten sphincterotomes were used in 110 patients (mean, 11 uses). Mechanical failure occurred primarily at the wire-insertion port, resulting in progressive friction with reuse. There were neither electrical nor infectious complications associated with reuse. CONCLUSIONS: A reusable double-channel sphincterotome is available which can theoretically be reprocessed and sterilized without the manual cleaning step of the reprocessing process. Contingent upon both provider and patient, multiple reuse can be anticipated, and contingent upon purchase price and reprocessing costs, the potential for procedural cost savings is significant. PMID- 11396757 TI - Outcome of endoscopic sphincterotomy in patients with pain of suspected biliary or papillary origin and inconclusive cholangiography findings. AB - BACKGROUND AND STUDY AIMS: We prospectively studied the outcome of endoscopic sphincterotomy in symptomatic patients with elevated liver enzyme levels but no clear evidence of biliary pathology on transabdominal ultrasound and diagnostic endoscopic retrograde cholangiography (ERC). METHODS: 29 consecutive patients with biliary-type pain (two or more out of eight criteria), elevated liver enzyme levels and no evidence of gallstones or significant common bile duct dilatation were evaluated. Elevated bilirubin levels (up to 7.2 mg/dl) were found in 18 patients. The majority of patients (n = 21) had a gallbladder in situ. The findings from bile duct exploration following sphincterotomy were recorded, and pain (as measured by visual analogue scale) as well as laboratory findings was assessed. RESULTS: Wire-guided sphincterotomy was successful in all patients while uncomplicated pancreatitis occurred in one instance. In 16 patients (55%) there was macroscopic evidence of small stones (n = 2), sludge (n = 12) or both (n = 2) following bile duct exploration. In addition, microscopy showed bile crystals in all four patients who had no macroscopic findings. All four patients with elevation of pancreatic enzymes prior to treatment, and four of those eight patients with previous cholecystectomy, showed evidence of biliary pathology. The initial median pain intensity was 8 (range 1-10); 26 patients became pain-free within 3 months following endoscopic sphincterotomy. While 26 of 28 patients (93%) remained asymptomatic over a median follow-up period of 19 months (range 12 26), one died of an unrelated malignancy 6 months after therapy. CONCLUSIONS: Endoscopic sphincterotomy may be acceptable in patients with typical clinical presentation suggesting a papillary or biliary origin of pain without further diagnostic work-up. Contrary to expectations, diagnostic ERC was insensitive in detection of the biliary etiology of symptoms in this selected group of patients. PMID- 11396758 TI - Long-term follow-up of percutaneous transhepatic therapy (PTT) in patients with definite benign anastomotic strictures after hepaticojejunostomy. AB - BACKGROUND AND STUDY AIMS: Percutaneous transhepatic therapy (PTT) is a promising minimally invasive procedure for benign stenosis of the anastomosis after hepaticojejunostomy. In this prospective study, the effectiveness and safety of this technique were investigated. PATIENTS AND METHODS: Between October 1995 and May 2000 34 consecutive patients were referred for treatment of symptomatic cholestasis due to anastomotic strictures after hepaticojejunostomy. In all patients percutaneous transhepatic cholangioscopic (PTCS) drainage and bougienage of the stenosis up to 16 Fr were performed. Associated bile duct stones were fractured using PTCS laser lithotripsy and removed into the jejunum. Afterwards, patients received transhepatic drainage for 3 months initially. The tubes were replaced in case of persistent strictures every 3 months up to 1 year. Patients in whom treatment failed underwent surgery or received biliary metal stents, depending on risk factors and individual anatomy. RESULTS: The procedure was performed in 34 patients (mean age 57 +/- 15) with cholestasis (alkaline phosphatase 691 +/- 485 U/l, bilirubin level mean 3.2 +/- 3.1 mg/dl). The transhepatic tube was successfully positioned into the right hepatic bile duct (n = 25), into the left (n = 3), or into both (n = 3) after 4 +/- 1 sessions, except in two patients in whom an external drainage was used and another patient in whom the procedure had to be stopped due to a bleeding complication. In 14 patients bile duct stones were successfully treated by PTCS laser lithotripsy before the placement of a transhepatic tube. The 30-day morbidity and mortality rates were 23.5% and 0% respectively. In 23 patients, the transhepatic tube could be removed after 212 +/- 122 days, with no evidence of cholestasis during a further follow up of 736 +/- 479 days. Four patients received metal stents because of persistent strictures after transhepatic intubation. Surgery had to be performed in a total of five patients, because of recurrent bile duct stones or recurrent strictures, in one patient with previous implantation of a metal stent, and in two patients with bile duct disconnection. Two patients died, one 1.5 years after surgery and one 427 days after metal stent implantation. None of these cases was related to the procedure. In two patients, the transhepatic tube is still in situ. CONCLUSIONS: Percutaneous transhepatic treatment of anastomotic strictures after hepaticojejunostomy is safe and highly effective in achieving internal biliary drainage. Temporary transhepatic intubation seems to be a promising minimally invasive alternative to surgery. PMID- 11396759 TI - Is endoscopic balloon dilation for removal of bile duct stones associated with an increased risk for pancreatitis or a higher rate of hyperamylasemia? AB - BACKGROUND AND STUDY AIMS: We studied the rate of pancreatitis and asymptomatic hyperamylasemia after endoscopic balloon dilation (EBD) and endoscopic sphincterotomy (EST) for removal of bile duct stones. PATIENTS AND METHODS: Patients with bile duct stones of all sizes were randomly allocated to undergo EBD (8-mm dilation balloon) or EST. Pancreatitis was defined as epigastric pain combined with at least a threefold rise in serum amylase at 24 hours after the endoscopic retrograde cholangiopancreatography (ERCP). Asymptomatic hyperamylasemia was defined as a threefold rise in serum amylase without epigastric pain. RESULTS: There were 180 patients (67 men, 113 women; mean age 67, SD 16.2) available for analysis. Complete stone removal after a single ERCP was achieved in 82 (88%) of 93 EBD patients and in 81 (93%) of 87 EST patients (P = 0.38). Mechanical lithotripsy was used more frequently in the EBD group (31% vs. 13%, P = 0.005). Early complications occurred in 16 EBD patients (17%) and in 19 EST patients (22%) (P = 0.46). Pancreatitis was observed in seven patients in each group (8%). Logistic regression identified no significant predictors for the occurrence of pancreatitis. Asymptomatic hyperamylasemia occurred in 21 EBD patients (23%) vs seven EST patients (8%) (P = 0.008). Logistic regression identified EBD as the only significant predictor for asymptomatic hyperamylasemia: odds ratio 2.9 (95% confidence interval (CI) 1.1 to 7.3, R2 = 0.02). CONCLUSIONS: We did not observe a difference in the rate of pancreatitis between EBD and EST. Asymptomatic hyperamylasemia was observed more frequently after EBD. Although asymptomatic hyperamylasemia is not a clinical entity, this finding may indicate that EBD causes more irritation of the pancreas than EST. PMID- 11396760 TI - Endoscopic sclerosis versus cyanoacrylate endoscopic injection for the first episode of variceal bleeding: a prospective, controlled, and randomized study in Child-Pugh class C patients. AB - BACKGROUND AND STUDY AIMS: Despite the recognized efficacy of sclerotherapy and elastic band ligation in controlling variceal hemorrhage, the results of endoscopic treatment in Child-Pugh class C patients remain poor. The aim of this prospective, controlled, and randomized study was to compare conventional sclerotherapy with injection of the tissue adhesive N-butyl-2-cyanoacrylate in controlling the first episode of rupturing of esophageal varices. PATIENTS AND METHODS: From January 1994 to June 1997, 36 consecutive Child-Pugh class C cirrhotic patients were admitted with an initial episode of esophageal variceal bleeding. They were randomly assigned to receive sclerotherapy with a 3% ethanolamine oleate solution (group 1, 18 patients) or injection of tissue adhesive (group 2, 18 patients). Episodes of recurrent bleeding were managed after the randomization procedure. After bleeding had been controlled, patients in both groups received weekly sessions of conventional sclerotherapy to eradicate any remaining esophageal veins. RESULTS: The patients in the two treatment groups had similar characteristics on entry into the study. More than 80% of the patients were admitted with moderate or severe hemorrhage. Approximately half of them presented with active bleeding during the index endoscopy. Early recurrent bleeding was observed in ten of the 18 patients in group 1 (55.6%) and in two of the 18 in group 2 (11.1%; P = 0.01). The hospital mortality rates were 72.2% in group I and 33.3% in group II (P = 0.04). CONCLUSION: These findings support the view that cyanoacrylate injection is superior to conventional sclerosis for controlling esophageal variceal bleeding in Child-Pugh class C patients. It is also highly probable that the better bleeding control achieved using the cyanoacrylate tissue adhesive treatment led to a lower hospital mortality rate. PMID- 11396761 TI - Is histological investigation of polyps always necessary? AB - BACKGROUND AND STUDY AIMS: To assess whether polyp histological type can be predicted by patient characteristics and endoscopic polyp findings. PATIENTS AND METHODS: 1681 polyps in 494 patients were categorized as advanced adenomas (villous component or severe dysplasia or early cancer) or insignificant polyps. Chi-squared tests were used to analyze whether polyp histological type could be predicted based on patient age (< 60 vs. > or = 60 years), gender, family history of colon polyps or cancer, presence of anemia, polyp size (< or = 5 mm vs. > 10 mm), and location (left- vs. right-sided). RESULTS: Insignificant polyp histology (n = 1337) correlated with patient age < 60 years (P = 0.0026), lack of anemia (P< 0.0001), polyp size < or = 5 mm (P < 0.0001), and right-sided location (P= 0.0058). Stepwise inclusion of these parameters demonstrated that the association of a < or = 5 mm right-sided polyp in a patient < 60 years yielded the highest combined predictive value (96.2%) for an insignificant polyp. Conversely, age > or = 60 years, presence of anemia, polyp size > 10 mm, or left-sided location, as single or combined parameters, demonstrated a maximum predictive value of only 75.4% for an advanced adenoma. CONCLUSIONS: A small right-sided polyp in a young patient is associated with a small risk (3.8 %) for advanced adenomatous tissue, indicating that histological investigation of such a polyp might not always be necessary. However, the recent recognition of flat adenomas and "mini" de novo colon carcinomas in the European population also may limit the usefulness of small polyp diameters in the exclusion of severe polyp histology. PMID- 11396762 TI - Percutaneous endoscopic gastrostomy in small medically complex infants. AB - BACKGROUND AND STUDY AIMS: Percutaneous endoscopic gastrostomy (PEG) is an established procedure for pediatric patients; however, there is still relatively little information on its feasibility and safety in very small infants. The aim of this study was to investigate the safety of percutaneous endoscopic gastrostomy in infants weighing less than 3.5 kg. PATIENTS AND METHODS: The charts of 26 infants weighing less than 3.5 kg who received PEGs were retrospectively reviewed. RESULTS: At the time of gastrostomy insertion the mean weight was 3 kg and the mean age was 2.3 months. This population of infants carried multiple diagnoses including lung disease of prematurity, swallowing dysfunction, chromosomal abnormality, structural facial anomaly, neurological deficit and congenital heart disease. Infants received either a 14- or 15-Fr percutaneous endoscopic gastrostomy tube under general anesthesia. All 26 procedures were successfully completed. Two infants (7.6%) developed a pneumoperitoneum during the procedure which required intervention. Two infants (7.6%) were conservatively treated with oral antibiotics for mild skin erythema and one infant (3.8%) required intravenous antibiotics for cellulitis of the stoma site. There were no other complications. To date, 16 of the gastrostomy tubes (61.5%) have been removed by traction without complication. CONCLUSIONS: PEGs can be safely placed in very small, medically complex infants. Pneumoperitoneum, which is a common but usually insignificant occurrence in adults and children during PEG placement, may require intervention in the small infant. PMID- 11396763 TI - Endoscopic mucosal resection and full thickness resection with complete defect closure for early gastrointestinal malignancies. AB - BACKGROUND AND STUDY AIMS: We developed a new endoscopic technique, the endoscopic full-thickness resection (EFTR) and endoscopic complete defect closure (ECDC), for the treatment of early gastrointestinal malignancies. PATIENTS AND METHODS: Two rectal and one duodenal carcinoid were treated by EFTR with ECDC and resectability, complications and pathological findings were evaluated. RESULTS: In all cases, the lesion was completely resected without serious complications. The histology of the specimen also confirmed complete resection of the tumor. CONCLUSIONS: This technique has great potential for the treatment of early gastrointestinal malignancies. Preliminary results have shown that early malignant gastrointestinal lesions can be safely and completely resected by EFTR with ECDC. PMID- 11396764 TI - Efficacy of a distal attachment in endoscopic resection of colorectal polyps situated behind semilunar folds. AB - BACKGROUND AND STUDY AIMS: It is difficult, dangerous, and time consuming to resect colorectal polyps situated behind the semilunar folds. The aim of this study was to examine the efficacy of a distal attachment in the endoscopic resection of these polyps. PATIENTS AND METHODS: The study group consisted of 11 patients, each with a colorectal polyp situated behind a semilunar fold, and of which the base was invisible without the use of a distal attachment. With the attachment fitted to the distal end of the endoscope, we pressed the semilunar fold situated at the anal side of the polyp so that the polyp, including its base, could be brought within the visual field. Maintaing the optimal position, we snared, ligated, and resected all polyps by electrocautery. RESULTS: The polyps were of sessile type in eight patients and pendunculated type in three, with a diameter ranging from 7 to 18 mm (mean 11 mm). Although the base of the polyp was invisible under conventional observation, pressing the fold brought good visualization. The optimal position for snaring the polyp was secured by fine endoscopic adjustment, and the polyps were easily resected in all patients. CONCLUSION: We conclude that the distal attachment is a simple but very valuable tool in the endoscopic resection of colorectal polyps situated behind the semilunar folds, and recommend its routine use in both diagnostic and therapeutic colonoscopy. PMID- 11396765 TI - Chronology of histological changes after band ligation of esophageal varices in humans. AB - BACKGROUND AND STUDY AIMS: While the histological effects of endoscopic sclerotherapy in humans have been extensively described, the effects of endoscopic ligation have been reported in only two cases. The purpose of this study was to reconstruct the chronological sequence of histological changes after ligation of esophageal varices. PATIENTS AND METHODS: Autopsy specimens from six patients who received ligation of varices from nine hours to 22 months ante mortem were evaluated for gross and microscopic changes. RESULTS: Early after ligation, the appearance was that of a polyp with its base compressed by the band. Variceal thrombosis was seen on day 2. Varying degrees of ischemic necrosis of the polyp were present on days 0-5. If the bands did not remain in situ for two days (premature loss), necrosis of the polyp and dilated variceal vessels were seen. On day 22, superficial ulcers were observed. After complete healing, fibrosis was seen in the submucosa. CONCLUSIONS: The changes seen in the present study are similar to those described in animals. The delay in ulcer healing, compared with the gross changes reported during follow-up endoscopic examinations, may be related to the severity of the underlying illness and the compromised immune status of patients in the present series. PMID- 11396766 TI - Neutrophil activation and hyperamylasaemia after endoscopic retrograde cholangiopancreatography: potential role for the leukocyte in the pathogenesis of acute pancreatitis. AB - BACKGROUND AND STUDY AIMS: Hyperamylasaemia occurs in up to 60% of patients following endoscopic retrograde cholangiopancreatography (ERCP), and in a small proportion of patients (1-5%) acute pancreatitis may develop. We evaluated the role of the neutrophil in post-ERCP hyperamylasaemia and acute pancreatitis by measuring circulating CD11b adhesion receptor expression--an indicator of leukocyte activation. PATIENTS AND METHODS: A total of 43 patients undergoing elective ERCP were studied. Peripheral blood measurements of amylase activity and neutrophil CD11b content (by flow cytometry) were made immediately before ERCP (baseline), and at 2 and 24 hours after the procedure. RESULTS: ERCP induced an increase in amylase level above baseline in 41 of 43 patients. The 2-hour and 24 hour post-ERCP amylase levels were directly related (R = 0.9, P < 0.01). Baseline CD11b receptor status was positively correlated with post-ERCP amylase activity (R = 0.4, P < 0.05), and this relationship was stronger when pancreatography had been performed (R = 0.67, P < 0.01). Three patients (7%) developed clinical acute pancreatitis, with post-ERCP amylase levels persistently elevated above 1000 IU/l. Multiple linear regression identified CD11b expression as the most significant explanatory variable for amylase level after ERCP (multiple R = 0.74, P < 0.01). CONCLUSIONS: The findings from this pilot study indicate an association between neutrophil activation and hyperamylasaemia following ERCP, and suggest a role for this leukocyte in the pathogenesis of pancreatitis. Further study of neutrophil characteristics may allow identification of individual susceptibility to ERCP-induced pancreatic injury. PMID- 11396767 TI - Clinical relevance of the small colorectal polyp. PMID- 11396768 TI - Enteroscopic cyanoacrylate sclerotherapy of jejunal and gallbladder varices in a patient with portal hypertension. AB - Bleeding from varices outside the gastroesophageal region is a rare, but regularly reported complication of portal hypertension. The treatment differs from the management of esophageal and gastric varices. We present here a report on the diagnosis and treatment of bleeding jejunal and gallbladder varices in a man with portal hypertension caused by chronic calcifying pancreatitis. The patient was suffering from recurrent, frequent, and massive gastrointestinal bleeding from varices at the anastomotic area of a cholecystojejunostomy. For diagnostic purposes, we carried out percutaneous Duplex ultrasonography and push enteroscopy with the Doppler technique. The treatment of varices in this area is traditionally surgical. This is the first report of enteroscopic sclerotherapy being successfully carried out using cyanoacrylate to treat hemorrhage from jejunal and gallbladder varices. No clinical signs of gastrointestinal bleeding were observed during a follow-up period of seven months. PMID- 11396769 TI - Level of sedation needed for colonoscopy in an individual patient, and how it should be predicted. PMID- 11396770 TI - Self-expanding metal stents or conventional prostheses for palliation of dysphagia? PMID- 11396771 TI - Foreign body: an unreported association with mucosectomy scar. PMID- 11396772 TI - Anal leukoplakia: an unusual case of anal stenosis. PMID- 11396773 TI - Dysphagia caused by Plummer-Vinson syndrome. PMID- 11396774 TI - Endosonography probe-guided endoscopic resection of small flat rectal carcinoid tumor using band ligation technique. PMID- 11396775 TI - A pill turned into a foreign body in a patient in a hurry. PMID- 11396776 TI - Influence of inositolhexaphosphori acid (phytic acid) on the copper distribution in tissues and the excretion of copper in rats. AB - The aim of the present study was to investigate the effect of the administration of phytic acid on copper (Cu) concentrations in several different rat tissues. The animals used were divided into three groups: Group A (received a diet supplemented with 2% phytic acid), group B (received a diet supplemented with 10% phytic acid) and group C (control). At the end of the experiment, the animals were sacrificed and the concentration of copper was determined in the different tissues. Phytic acid significantly increased Cu concentration in the duodenum of the animals of both groups as well as in the lungs and blood of the animals of group A. The copper concentration was also increased in the uterus and bone of the animals of group B. On the other hand, the stomach copper concentration of the animals of both groups, the heart and lung copper concentrations of the animals of group B as well as the jejunum, colon and hair copper concentrations of the animals of group A were significantly decreased. Copper excretion through feces was significantly decreased in the animals of both groups, while the excretion through urine was not significantly affected by the administration of phytic acid. In conclusion, the administration of phytic acid can produce translocation and/or elimination of copper in various tissues of rats. PMID- 11396777 TI - Absorption and retention of selenium from shrimps in man. AB - This study was undertaken to evaluate the bioavailability of selenium in shrimps, a possible good source of selenium, by measurements of the absorption and retention of selenium and the effects on plasma selenium concentration and glutathione peroxidase activity. Twelve healthy young subjects (9F and 3M) received 100 g of shrimps each day for six weeks in addition to their habitual diet. In the third week of the study a balance period was inserted in which the subjects received all food from the department and collected faeces and urine over 5 days. Blood samples were collected at commencement of the study, after 2, 4, and 6 weeks. The selenium intake increased from 39.4 +/- 15.3 microg/d to 127 +/- 5.5 microg/d with the addition of shrimps. The apparent absorption of selenium from shrimps was 83 +/- 4%. Faecal and urinary selenium excretion was 32.5 +/- 17.0 microg/d and 21.2 +/- 9.0 microg/d, respectively and the total retention of selenium was 3.1 +/- 1.1 mg. Plasma selenium concentrations were 95.2 +/- 9.7 microg/L and 101.5 +/- 9.7 microg/L before and after six weeks of shrimp intake, respectively (p<0.05). Plasma and erythrocyte glutathione peroxidase activities were not influenced by shrimp intake. Thus, despite the high absorption and retention, plasma selenium concentrations were only moderately affected by an increase in selenium intake of about 100 microg/d in the chemical forms found in shrimp. Whether the accumulation of selenium from shrimps in tissues may represent a potential hazard is to be further investigated. PMID- 11396778 TI - Element concentrations and cataract: an experimental animal model. AB - The determination of inorganic ions in cataractous human lenses has been the subject of several investigations; nevertheless, few studies have been concerned with trace element contents in lenses, and data are sometimes contradictory. An animal experimental model of induced cataract is here proposed with the aim of evaluating the changes of Ca, Na, K, Cu and Zn concentrations. The cataract was produced by an Nd:YAG Laser treatment of the right eye of sexteen male rabbits. The determination of the elements was performed by atomic absorption spectrometry (both flame and flameless methods) after an acid digestion of samples. Compared with the results obtained in left lenses used as a control (Ca 14.4+/-5.7 mg/kg d.w.; Na 1.3+/-0.5 g/kg d.w.; K 9.9+/-1.1 g/kg d.w.; Cu 0.24+/-0.09 mg/kg d.w.; Zn 24.8+/-2.3 mg/kg d.w.), the mean concentration values of opaque lenses showed some significant changes for Ca, Na, and Cu (Ca 123.7+/-106.6 mg/kg d.w.; Na 4.5+/-4.3 g/kg d.w; Cu 0.43+/-0.21 mg/kg d.w.). Potassium showed a tendency to decrease, and zinc to increase. Positive correlations were found between calcium and sodium both in controls (r=0.73, p<0.001) and in treated lenses (r= 0.87, p<0.0001). An inverse correlation between Ca and K confirmed the tendency of potassium to decrease. PMID- 11396779 TI - Urinary selenium and iodine during pregnancy and lactation. AB - The New Zealand environment is low in selenium and iodine, and is therefore ideally suited for the study of these anionic trace elements. The aim of this study was to determine urinary excretion of selenium and iodine during pregnancy and postpartum as part of an investigation of the influence of pregnancy and lactation on selenium metabolism in women of low selenium status. In a double blind placebo-controlled study, 35 women in the earliest stages of pregnancy and 17 non-pregnant women were recruited in Dunedin, New Zealand. Eighteen pregnant women received 50 microg selenium as L-selenomethionine, while the others received a placebo daily during pregnancy and 12 months postpartum. The non pregnant women received the supplement, serving as a positive control. Blood samples and twenty-four hour urine samples were collected monthly during pregnancy and at 3, 6, and 12 months postpartum for analysis of selenium and iodine. Selenium content in plasma and urinary excretion of selenium fell during pregnancy; however, total excretion of selenium was greater during pregnancy than postpartum. Urinary iodine excretion was much lower than reported previously in New Zealand. Due to large intra- and inter-subject variability, no trends in iodide excretion were observed. Factors which influence urinary excretion of selenium include dietary intake, but more closely, plasma concentrations of selenium (which is probably related to total selenium pool), creatinine excretion and therefore lean body mass, and glomerular filtration rate. The exact mechanism and sequence of events remains unclear and future studies incorporating new speciation techniques are necessary. PMID- 11396780 TI - Plasma selenium in preterm and term infants during the first 12 months of life. AB - The goal of the present study was to prospectively assess the plasma selenium (Se) concentrations of term and preterm infants during the first year of life in relation to gestational age and nutrition. Blood specimens were collected from orally formula-fed preterm infants (gestational age < 32 weeks, birth weight < 1500 g): 1.) in hospital and 2.) corrected for gestational age parallel to healthy term breast and formula-fed infants at the ages of 1, 4 and 12 months. All infants were fed according to a standardized nutritional concept, solids and follow-up formula were introduced at the age of 4 months. Plasma selenium in preterm infants in hospital was 11.7 (6.5-20.8) microg/l and 11.6 (8.8-16.7) microg/l at 4 weeks corrected for gestational age. At the age of 4 months plasma selenium was still significantly lower than in the other groups: Preterm infants: 17.1 (10.4-30.5) microg/l; formula-fed term infants: 31.3 (24.3-47.5) microg/l; breast-fed term infants: 45.6 (27.1-65.1) microg/l). The levels of breast-fed infants were significantly higher than those of both formula-fed groups up until the introduction of solids. Preterm infants had significantly low plasma selenium levels up until a postnatal age of at least 6 months. The levels were lower than those of term infants fed an identical unsupplemented infant formula during the first 4 months of life. These data support routine monitoring in hospital and selenium supplementation of preterm infants, preferably in hospital before discharge. PMID- 11396781 TI - Aluminum balance in intensive care patients. AB - Aluminum (Al) is a well known contaminant of intravenous solutions. The aim of the present study was the estimation of the aluminum load in patients of an intensive care unit (ICU). 15 patients with normal renal function took part. The study period was 15 days. Al was measured in serum, 24 h-urine and 132 samples of parenterals. Daily Al doses were recorded. Al balance was calculated on the basis of the iatrogenic Al dose and renal Al excretion. Al analysis was performed by graphite furnace atomic absorption spectrometry (AAS) with Zeeman background correction under careful quality control. Solutions with Al levels >100 microg/l were: calcium salts, additives for parenteral nutrition solutions, antibiotics, acetylcysteine, triflupromazine, catecholamines and colloids. The Al content of solutions for parenteral nutrition ranged from 4.3 to 69 microg/l. Al doses amounted to 46-456 (median 119) microg/d, equivalent to 0.7 to 6.5 (median 1.7) microg/kg b.w. Renal Al excretion ranged from 10.5 to 723.1 microg/d (median 53 microg/d). These amounts partly exceeded the maximal dose (2 microg/kg b.w. per day), recommended by ASPEN/ASCN. Despite of the highly elevated renal Al excretion the median serum concentration of Al was only moderately increased (6.1 microg/l; range: <1.5 to 23.6 microg/l). However, calculations on the basis of the iatrogenic Al dose and renal Al excretion resulted in a net Al uptake (median) of 61 microg/d (maximum: 291 microg/d). Al amounts of this magnitude must be considered potentially harmful in ICU patients, especially with impaired renal function. Parenteral therapy resulted in a considerable Al dose with a positive Al balance in ICU patients. Threshold values for Al contamination of parenterally administered drugs and solutions should be established. PMID- 11396782 TI - Plasma status of selected minerals in hypertensive men with and without insulin resistance. AB - The altered plasma statuses of selected minerals (Ca, Mg, Cu, Zn) have been noted in a cluster of insulin resistance syndromes, including hypertension and diabetes mellitus. The differences in plasma values of these minerals in hypertensive men with and without insulin resistance, as evaluated by an insulin suppression test, were investigated. The results showed that the plasma values of determined minerals at fasting, 2 h after an oral glucose challenge, and after the insulin suppression test did not markedly differ between hypertensive subjects with and without insulin resistance. However, hypertensive subjects had significantly lower plasma Ca values at fasting and 2 h after an oral glucose load, and higher fasting plasma Zn values, than normotensive controls. Hypertensive subjects also had higher steady-state plasma glucose values, higher Zn and lower Mg and Cu values after the insulin suppression test, when compared with controls. The present study suggests that altered plasma status of selected minerals in hypertension cannot be totally ascribed to the co-exhibition of insulin resistance. PMID- 11396783 TI - The potencial role of rare earths in the pathogenesis of interstitial lung disease: a case report of movie projectionist as investigated by neutron activation analysis. AB - A 60-year-old male subject who worked as a movie projectionist and who was exposed for 12 years to rare earths (RE) containing dusts from cored arc light carbon electrodes was investigated. Chest X-ray films and pulmonary function tests showed an interstitial lung disease, emphysema and a severe obstructive impairment with marked decrease of carbon monoxide diffusion capacity. The histological examination of a transbronchial biopsy confirmed the diffuse interstitial lung fibrosis. Neutron activation analysis (NAA) of the biopsy showed concentrations of cerium (Ce), lanthanum (La), neodimium (Nd), samarium (Sm), terbium (Tb) and ytterbium (Yb) which were high compared to the corresponding elements in the transbronchial biopsies of 5 unexposed subjects as a control group. Thorium (Th) (which is generally present as an impurity of the RE compounds) was also determined in order to estimate the radiation dose in the lung of the worker. On the basis of the clinical observations, of the analytical results by neutron activation analysis of RE and of the presence of Th in the transbronchial biopsy, as well as of the differential diagnosis, which tended to exclude other occupational or non-occupational lung diseases, a relation between the observed interstitial lung fibrosis and occupational exposure to RE is highly probable. PMID- 11396786 TI - Review of publications. PMID- 11396784 TI - Iron overload can induce mild copper deficiency. AB - Dietary copper in the U.S. often is lower than that proved insufficient for men and women under controlled conditions. Iron overload can have adverse effects on copper nutriture and can produce cardiac disease in people. The hypothesis that iron can interfere with copper utilization to produce adverse effects related to cardiovascular function was tested. Rats were fed a diet high in iron and marginal, but not deficient in copper for comparison with similar diets containing iron at the recommended amount. Copper and iron were measured by atomic absorption spectroscopy; cholesterol was measured by fluorescence, ceruloplasmin was measured by oxidase activity and hematology was done by an automated cell counter. When dietary copper was 2.0 mg/kg of diet, high iron decreased (p<0.008) cardiac and hepatic copper, plasma copper and ceruloplasmin, and increased (p<0.02) cardiac weight, hepatic iron and plasma cholesterol. When dietary copper was increased to 2.5 mg/kg, copper in heart and plasma decreased (p<0.04) and hepatic iron increased (p=0.001) with high iron but other effects disappeared. No harmful changes in hematology, such as hematocrit, mean corpuscular volume, etc. were found. High iron increased the dietary copper requirement of the animals. People with iron overload may benefit from copper supplementation, particularly if they habitually consume a diet low in copper. PMID- 11396785 TI - Application of inductively coupled plasma sector field mass spectrometry for elemental analysis of urine. AB - An analytical method using double focusing sector field inductively-coupled plasma mass spectrometry (sector field ICP-MS) for rapid simultaneous determination of 42 elements in urine is described. Sample preparation consisted of 20-fold dilution with 0.14 mol/l nitric acid in ultrapure water. The importance of controlling possible contamination sources at different sample preparation and analysis stages in order to achieve adequate method detection limits (DL) is emphasized. Correction for matrix effects was made using indium and lutetium as internal standards. Different approaches for accuracy assessment in urine analysis are evaluated. Additional information on trace element concentrations in a urine reference material is given. Between-batch precision was assessed from the analysis of separately prepared aliquots of the reference material and was better than 10% RSD for 32 of the elements. The robustness of the procedure was tested by analysis of about 250 samples in one analytical run lasting more than 50 hours. A statistical summary of results for 19 urine samples from non-exposed subjects is presented. For a majority of the elements tested concentrations were higher than the detection limit of the method. PMID- 11396787 TI - On the new anatomical nomenclature. AB - The present paper is concerned with the linguistic aspect of the new anatomical nomenclature (Terminologia Anatomica 1998). Orthographic, morphological, syntactic, lexical, and terminological comments are presented. In the authors' opinion, shortcomings might have been effectively avoided by cooperation with linguists. PMID- 11396788 TI - Upregulation of vasoactive intestinal polypeptide (VIP) and calcitonin gene related peptide (CGRP) expression in stellate ganglia of children with congenital cardiovascular lesions. AB - The distribution patterns of vasoactive intestinal polypeptide (VIP) and calcitonin gene-related peptide (CGRP) immunoreactivities (IR) in stellate ganglia of human neonates and infants with congenital heart and vascular lesions were investigated by the method of indirect immunohistochemistry. The results demonstrated upregulation of VIP and CGRP expression in principal ganglionic neurons independently of the type of lesion. It is suggested that the activation of neuropeptide synthesis in stellate ganglia is a compensatory reaction of ganglionic neurons in response to congenital cardiovascular lesions, in regulation of heart contractility, and as a trophic influence on the ischemic myocardium. Hypoxia is the main inducing factor for the upregulation of VIP and CGRP expression in sympathetic neurons. PMID- 11396790 TI - Morphological diversity of dying cells during regression of the human tail. AB - During normal human development a number of transient structures form and subsequently regress completely. One of the most prominent structures that regress during development is the human tail. We report here a histological and ultrastructural study of cell death in the cranial and caudal (tail) parts of the neural tube in 4 to 6-week-old human embryos. Initially, the human tail is composed of tail bud mesenchyme which differentiates into caudal somites, secondary neural tube, notochord and tail gut. Later on, these structures gradually regress by cell death. During the investigated period, we observed two morphologically distinct types of dying cells. The well-described apoptotic type of cell death was observed only in the cranial neural tube that forms during primary neurulation. The other type of cell death characterized by necrotic morphology was observed in the tail mesenchyme and in the caudal neural tube that forms during secondary neurulation. This morphological diversity suggests that besides differences in origin and fate there are different mechanisms of developmental cell death between two parts of the human neural tube. We can speculate that the apoptotic type of cell death is associated with the precise control of cell numbers and that the other morphologically distinct type of cell death is responsible for the massive removal of transitory structures. PMID- 11396789 TI - Distribution of vasoactive intestinal polypeptide-, calcitonin gene-related peptide-, somatostatin- and neurofilament-immunoreactivities in sympathetic ganglia of human fetuses and premature neonates. AB - The distribution patterns of vasoactive intestinal polypeptide (VIP), calcitonin gene-related peptide (CGRP), somatostatin (SOM) and neurofilament (NF) immunoreactivities (IR) were studied in the stellate ganglia of human fetuses and neonates at 24-26 weeks gestation. Sizeable populations with some quantitative variations of VIP-, CGRP- and SOM immunoreactive nerve cells were detected in all ganglia studied. In marked contrast, neurofilament expression was down-regulated. The upregulation of VIP, CGRP and SOM expression suggested their inductor effect on growth and differentiation neurons as well as on the development of their neurotransmitter properties. The main neuropeptides-inducing factor of sympathetic ganglia in human prenatal ontogenesis may be considered as a relative hypoxia. PMID- 11396791 TI - Immunohistochemical detection of estrogen receptor alpha in articular chondrocytes from cows, pigs and humans: in situ and in vitro results. AB - Clinical observations suggest that estrogens are involved in the pathogenesis of postmenopausal osteoarthritis, but only little is known about the influence of these hormones on articular cartilage cells. The effect of estradiol is mediated by estrogen receptors alpha and beta. The goal of the present study was to search for estrogen receptor alpha in articular tissue from cows, pigs and humans by immunohistochemistry to form a basis for in vitro studies. In addition, we also tried to detect estrogen receptor alpha in cultivated articular chondrocytes from cows and bulls under certain culture conditions. Estrogen receptor alpha is detected by the use of antibody 13H2 in articular chondrocytes from cows, bulls, pigs and humans. Chondrocytes are physiologically exposed to reduced oxygen tension. In isolated articular chondrocytes from cows and bulls incubated either with 21% O2 or with 5% O2 positive cells were also found. These positive results therefore encourage testing the influence of estradiol on cultivated articular cartilage cells in these species under different culture conditions. PMID- 11396793 TI - Topography of muscle spindles and Golgi tendon organs in shoulder muscles of "Monodelphis domestica". AB - The topography of muscle spindles and Golgi tendon organs in the rotator cuff and surrounding shoulder muscles of a small laboratory marsupial (monodelphis domestica) were studied using light microscopy of serial sections. The shoulder joint of monodelphis has a large degree of freedom of movement allowing this animal to use the upper extremities for a wide range of activities like climbing and manipulating food. Thus, similar to the situation in man the shoulder joint is mainly secured by muscles. Silver stained serial paraffin sections were examined under the light microscope and the distribution of muscle spindles and Golgi tendon organs was reconstructed using three-dimensional image processing. In the two animals examined 113 and 131 muscle spindles respectively were found within the 4 rotator cuff muscles. In addition, 76 and 40 Golgi tendon organs respectively were seen at the musculo-tendinous junctions of these muscles preferentially close to the insertion at the humerus head. Also the surrounding shoulder muscles contain both muscle spindles and Golgi tendon organs in large numbers, but the ratio of Golgi tendon organs per muscle spindle appears to be lower. Number and localization of muscle spindles and Golgi tendon organs suggest, that these receptors are important for both reflex control of shoulder muscle tone as well as monitoring of static position and movement in the shoulder joint. PMID- 11396792 TI - Ultracytochemistry of glycosaminoglycans in the canine knee synovium. AB - The accurate localization and nature of glycosaminoglycans (GAGs) in the canine knee synovium were studied by ultracytochemical methods that involved high or low iron diamine-thiocarbohydrazide-silver proteinate (physical development) staining in combination with enzyme digestion control procedures. The results obtained indicated that heparan sulfates and hyaluronan were present mainly in the plasma membrane of the B (fibroblast-like) cells. In contrast, the plasma membrane of the A (macrophage-like) cells showed negative reactions after the histochemical examination. Dermatan sulfates, chondroitin sulfates (A and/or C) and hyaluronan were localized in the extracellular matrix of the synovial intima, whereby dermatan sulfates were confined to the fibrous component, whereas chondroitin sulfates and hyaluronan were found in the interfibrous matrix. Heparan sulfate was the only notable GAG molecular species localized in the basement membrane of the capillary wall. It is obvious that differences in the quality and localization of glycosaminoglycans in the canine synovial tissue are of specific interest in understanding normal functions as well as pathological alterations of the knee synovium in mammals. PMID- 11396794 TI - Vascular endothelial growth factor gene expression in the human breast cancer cell line MX-1 is controlled by O2 availability in vitro and in vivo. AB - The vascular endothelial growth factor (VEGF) plays an important role in angiogenesis. Mediated by the hypoxia-inducible transcription factor HIF 1alpha/beta, a reduction in O2 tension (pO2) leads to increased VEGF gene expression in nonmalignant tissues. In tumor cells VEGF mRNA levels are often constitutively elevated. We examined pO2-dependent VEGF mRNA expression and VEGF protein formation in the human breast cancer cell line MX-1 in vitro and in vivo. For in vitro study MX-1 cultures were grown on dishes with a gas-permeable bottom to expose the cells to defined O2 concentrations (from 95% to 0%) for 4 h. Northern blot analysis showed significant VEGF mRNA in MX-1 cultures under normoxic conditions which was further increased by hypoxia. The amount of secreted VEGF was also elevated in hypoxic cultures. Western blot analysis revealed a correlation between the severity of hypoxia and HIF-1alpha protein amounts in the nucleus. Furthermore, DNA-binding activity of HIF-1 could be demonstrated by gel-shift assays. For in vivo study immunodeficient nude mice bearing MX-1 tumor transplants were exposed to inspiratory hypoxia (10% O2). Northern blot and immunohistochemical analyses of MX-1 tumor transplants showed that VEGF mRNA and VEGF protein levels were increased in mice 17 h after the induction of inspiratory hypoxia. Thus, pO2-dependence of VEGF gene expression can be maintained in cancer cells, even in vivo, which may be relevant in regard to therapeutic attempts to inhibit tumor angiogenesis by increasing tumor oxygenation. PMID- 11396795 TI - Endocytosis of ferritin and hemoglobin by the trabecular endocardium in swordtail, Xiphophorus helleri L. and platy, Xiphophorus maculatus L. (Poecilidae: Teleostei). AB - The cardiac atrium and ventricle of swordtail, Xiphophorus helleri L. and platy, Xiphophorus maculatus L., are spongious, consisting of muscle trabeculae covered by endocardial cells. The cardiac trabecular endocardium is able to take up and store large amounts of horse-spleen ferritin and bovine hemoglobin from the blood stream. No such uptake was registered in endocardial cells lining the cardiac valves, atrio-ventricular junction and ventriculo-bulbar junction. The trabecular endocardium in these species seems to be unable to accumulate latex beads or bovine myoglobin, cytochrome C and holotransferrin from the blood stream. It is proposed that the trabecular endocardium in these species is able to clear the blood stream of some types of waste macromolecules; i. e. this tissue may have a scavenger function. The present results indicate that the uptake of foreign ferritin in bony fish endocardium can be clearly demonstrated at the light microscopic level in deparaffined sections by means of acid ferrocyanide or Mallory solutions. A similar uptake of hemoglobin is demonstrated by means of Mallory stain. PMID- 11396796 TI - Mucosal microvasculature of the gastric pars nonglandularis and margo plicatus in the horse: a scanning electron microscopic study on corrosion casts. AB - The microvascularisation of the equine non-glandular gastric mucosa was investigated using corrosion casts for scanning electron microscopy. Specimens from 11 healthy horses were examined. Corresponding to the high incidence of gastric lesions in the margo plicatus, special attention was paid to the differentiation between the pars nonglandularis and the margo plicatus as a distinct area of the aglandular mucosa. In both areas, the blood vessels of the lamina propria mucosae were arranged in three vascular layers; i. e. I) a basal, II) an intermediate, and III) a subepithelial horizontal level. In the basal (I) and in the intermediate (II) layers the vascular supply was organised in arterial retia-rete arteriosum profundum, rete arteriosum subpapillare- and venous plexus plexus venosus profundus, plexus venosus subpapillare. Vertical interconnections integrated the layers into the vascular network of the entire lamina propria. The subepithelial (III) layer represented the blood vessels of all the individual connective tissue papillae in the lamina propria mucosae. Ansae capillares intrapapillares were found in the pars nonglandularis. In contrast, each of the papilla of the margo plicatus contained a "cone shaped" rete capillare intrapapillare. The thicker epithelium and lamina propria mucosae of the margo plicatus was therefore supplied by less numerous, but longer intrapapillary blood vessel systems. The typical vascular components of the margo plicatus may be considered to be one of several links in the etiological chain which characterises gastric mucosal lesions in the horse's stomach. PMID- 11396797 TI - Horseshoe lung: report on a new variant--"inverted" horseshoe lung--with embryological reflections on the formal pathogenesis of horseshoe lungs. AB - The term "horseshoe lung" is used to describe a rare congenital anomaly of the lungs that is characterized by the presence of a midline isthmus of pulmonary parenchyma connecting the posterobasal regions of the right and left lungs. Since the introduction of the term horseshoe lung in the 1960's, almost 40 cases have been reported in the literature. In all these cases, the right and left lungs were joined in their posterobasal regions, the situation resembling that found in horseshoe kidneys. Here we present a case of connection between the right and left lungs found during necropsy of a human fetus. In this case, a midline isthmus of pulmonary parenchyma covered by visceral pleura joined the apical regions of the right and left lungs behind the trachea and esophagus. This connection resulted in a "horseshoe"-shaped lung in which the "horseshoe" was turned by 180 degrees compared to classical cases of horseshoe lung. To the best of our knowledge, this is the first report on an "inverted" horseshoe lung. Embryological reflections on the formal pathogenesis of inverted and classical horseshoe lungs are presented. PMID- 11396798 TI - The different growth zones of the fetal foot. AB - Previous publications revealed no reliable data or models concerning the three dimensional ontogenesis of the lower extremity. Using the method of plastination histology in combination with 3D-computer-reconstructions we were able to produce exact, virtual 3D-specimens of 19 healthy fetal feet. The fetuses were aged between 9 to 38 weeks of gestation and age-dependently related to four defined age-groups. We compared these feet with the help of a new geometrical method. Thus, we obtained a kind of "slow-motion-picture" of the undisturbed three dimensional development of the fetal foot. Our results show that the human fetal foot has a desultory mode of growth and that growth priorities within the foot skeleton change dependent upon age and region. Though the growth of the fetal foot-skeleton is desultory, it is not disconnected. The result of this peculiar mode of growth is to create the foot arches and thus seems to be functionally oriented toward the human foot's specific purposes. PMID- 11396799 TI - Plastinated body slices for verification of magnetic resonance tomography images. AB - Plastinated body slices provide a higher resolution than photos taken from frozen objects as obtained for the "Visible Human Project". The impregnation with curable polymer gives those slices transparency. Since the thickness of a plastinated slice determines its optical resolution, very thin plastinated slices are necessary for comparison with MRT-images (MRIs) of high quality. The present technical note describes how very thin plastinated slices can be prepared by modification of the classical plastination technique and for comparison with MRI. Having labelled the object's planes recorded as MRIs, we, for the first time, plastinated 800 microm thick body slices superior in resolution to the corresponding MRIs, confirming and extending their structures. Thus, very thin plastinated body slices can be regarded as a helpful tool for radiological studies. PMID- 11396800 TI - Age-related changes in the ventricular system of the dog brain. AB - The cerebral ventricles represent cavities of the brain which are used for diagnostic purposes. Although a wide variety of age related changes have been described in the Central Nervous System (CNS) of many species, few studies about the effect of ageing on the canine brain have been published until now. However, to date there is no previous data concerning the ventricular system of dogs. The present study deals with the morphometry of the various parts of the cerebral cavities of the German Shepherd dog by means of Batson's casts and the possible changes which take place in the ventricular system with age and/or sex. In this study, two age groups were considered: young (2-5 years) and old (10-12 years). A total number of forty seven dogs (12 young males, 13 old males, 13 young females and 9 old females), weighing 34-42 kg, were used for experimentation. Our results describe the enlargement which takes place in the ventricular system with age, which is probably related to a general age-related atrophy of neural tissue. On the contrary, there are no remarkable changes related to sex. These age-related changes are similar to the best known changes which occur in the human species. Our data could corroborate the usefulness of the dog as a natural animal model for the study of normal ageing and neurodegenerative diseases. PMID- 11396801 TI - Accessory oval foramen. AB - We describe an opening with smooth walls in front and medial to foramen ovale which leads to an oblique canal directed towards the fossa pterygoidea. The canal was up to 2.3 mm long and opened near the root of the pterygoid process. We called this opening "foramen ovale accessorium", and found it in 48 of 124 anatomical specimens. The foramen was present in newborns and in those aged over 80 years old on one side but rarely on both sides of the sphenoid bone. Only in a single specimen were there two foramina side by side in front of the foramen ovale. In another case, the foramen ovale accessorium was present in the anterior wall of the canalis ovalis and pointed to the fossa pterygoidea. The localization and direction of the foramen ovale accessorium led us to the conclusion that it housed some of the separate rootlets for the chewing muscles, in this case for mm pterygoidei, mm tensor veli palatini and tensor tympani. The existence and contents of the foramen ovale accessorium is important in surgical interventions on the trigeminal nerve and/or ganglion Gasseri. PMID- 11396802 TI - Guidelines for approaching suspected peptic ulcer disease or Helicobacter pylori infection: where we are in pediatrics, and how we got there. PMID- 11396804 TI - Plasma nitrate concentration: a new diagnostic test in young infants with infectious gastroenteritis? PMID- 11396803 TI - Intestinal bile acid transport: biology, physiology, and pathophysiology. AB - Intestinal reabsorption of bile salts plays a crucial role in human health and disease. This process is primarily localized to the terminal ileum and is mediated by a 48-kd sodium-dependent bile acid cotransporter (SLC10A2 = ASBT). ASBT is also expressed in renal tubule cells, cholangiocytes, and the gallbladder. Exon skipping leads to a truncated version of ASBT, which sorts to the basolateral surface and mediates efflux of bile salts. Inherited mutation of ASBT leads to congenital diarrhea secondary to bile acid malabsorption. Partial inhibition of ASBT may be useful in the treatment of hypercholesterolemia and intrahepatic cholestasis. During normal development in the rat ileum, ASBT undergoes a biphasic pattern of expression with a prenatal onset, postnatal repression, and reinduction at the time of weaning. The bile acid responsiveness of the ASBT gene is not clear and may be dependent on both the experimental model used and the species being investigated. Future studies of the transcriptional and posttranscriptional regulation of the ASBT gene and analysis of ASBT knockout mice will provide further insight into the biology, physiology, and pathophysiology of intestinal bile acid transport. PMID- 11396805 TI - Clinical quiz. Sarcoidosis. PMID- 11396806 TI - Markers of cystic fibrosis-associated liver disease. PMID- 11396807 TI - Plasma nitrate concentrations in children with infectious and noninfectious diarrhea. AB - BACKGROUND: In patients with intact renal function and low dietary nitrate intake, plasma nitrate concentrations reflect endogenous nitric oxide production and are shown to be increased during inflammatory processes. The aim of this study was to compare plasma nitrate concentrations and hence endogenous nitric oxide production in children with infectious and noninfectious diarrhea and to determine whether plasma nitrate concentrations could serve as a discriminant test between acute and chronic diarrhea in children. METHODS: Three groups of patients were identified: 14 patients with acute gastroenteritis, 13 patients with chronic noninfectious diarrhea, and 14 patients with no evidence of gastrointestinal pathology and no underlying infectious process, who served as control subjects. Plasma nitrate concentrations were determined spectrophotometrically using the Greiss reaction before reduction to nitrite with a copper-coated cadmium column. RESULTS: Mean plasma nitrate concentrations were 405.3 micromol/L +/- 281.6 micromol/L (standard deviation) in patients with infectious diarrhea, 134.7 micromol/L +/- 77.0 micromol/L in patients with chronic diarrhea, and 54.1 micromol/L +/- 20.1 micromol/L in control subjects (F = 42.6, P < 0.0001; analysis of variance). Plasma nitrate concentrations were significantly higher in the infectious diarrhea group compared with the noninfectious diarrhea and control groups (Student-Newman-Keuls test, P < 0.5). CONCLUSIONS: Although an optimal cutoff concentration cannot be defined, plasma nitrate concentrations in excess of 300 micromol/L are suggestive of an infectious process whereas values less than 100 micromol/L are indicative of noninfectious diarrhea. PMID- 11396808 TI - Apoptotic epithelial cells in biopsy specimens from infants with streaked rectal bleeding. AB - BACKGROUND: Histologic studies of rectosigmoidal mucosal biopsies of infants with isolated blood-streaked stool have shown many eosinophils and revealed aggregates of small dark granules (nuclear dust). However, no description of the nuclear dust has been made for this condition and the nature of the nuclear dust has not been thoroughly investigated. We determined the characteristics of these particles in biopsies from infants with streaked rectal bleeding. METHODS: Nineteen infants who were younger than 6 months old and had isolated rectal bleeding were studied, as were six age-matched control infants. Rectosigmoidal mucosal biopsies were immunohistochemically assessed using anticarcinoembryonic antigen and macrophage-associated antibodies and examined for apoptotic cells by modified in situ TdT-mediated dUTP-biotin nick-end labelling. The number of apoptotic epithelial cells was compared between rectal bleeding and control groups. RESULTS: Immunohistochemistry showed that at least some of the nuclear dust consisted of apoptotic epithelial cells. Infants with rectal bleeding also showed nodular lymphoid hyperplasia (n = 16), abundant eosinophils (>20/high power field, n = 14) in the mucosa, and a significantly high number of apoptotic epithelial cells relative to the control group. Rectal bleeding disappeared at 6 month follow-up in 14 of 18 infants (one was lost to follow-up) who were fed a different milk formula or breast-fed (their mothers were restricted from having cow's milk and eggs). CONCLUSIONS: The high number of apoptotic epithelial cells in rectosigmoidal mucosal biopsies of infants with streaked rectal bleeding is probably caused by accelerated epithelial cell turnover and apoptosis. PMID- 11396809 TI - Assessment of hepatic function in cystic fibrosis by lidocaine metabolism. AB - BACKGROUND: To determine hepatic drug metabolism in patients with cystic fibrosis, as measured by monoethylglycinexylidide formation after lidocaine injection and indocyanine green (ICG) clearance. METHODS: The following study is a case-control study, which included 19 patients with cystic fibrosis and 13 control subjects. Serum monoethylglycinexylidide concentration was measured after intravenous injection of 1 mg/kg (maximum, 50 mg) lidocaine. Indocyanine green (0.5 mg/kg) was injected concomitantly, and absorbance (805 nm) of serum was measured over time to determine its volume of distribution, serum half-life, and hepatic blood flow. RESULTS: Monoethylglycinexylidide formation was decreased in patients with cystic fibrosis compared with controls (39.4+/-16.9 microg/L versus 70.3+/-45.7 microg/L, mean +/- SD, respectively, P < 0.02). Indocyanine green half-life (4.6+/-2.7 min versus 3.0+/-1.0 min), volume of distribution (8.6+/-5.5 L versus 8.3+/-3.4 L), and hepatic blood flow (10.9+/-5.9 ml x kg(-1) x min(-1) versus 7.4+/-2.0 ml x kg(-1) x min(-1)) were similar in both groups. CONCLUSION: Monoethylglycinexylidide formation after lidocaine injection is impaired in patients with cystic fibrosis. This impairment may have clinical implications when using hepatically metabolized medications in patients with cystic fibrosis. PMID- 11396810 TI - Nutritional zinc balance in extremely low-birth-weight infants. AB - BACKGROUND: Zinc is important for metabolism, cell growth, immunity, and defense against oxygen radicals. Extremely low-birth-weight (< 1000 g) infants have higher nutritional needs, but information on zinc is scarce. The authors performed nutritional balances in 10 infants with birth weights of 500 to 999 g and who were fed with fortified human milk. METHODS: The authors collected infant feces, urine, and blood and human milk samples during 72 hours at 7 and 12 weeks of age. Zinc concentration was measured by inductively coupled plasma-mass spectrophotometry, atomic emission spectrophotometry, and instrumental neutron activation analysis. RESULTS: Mean (SD) intake via human milk was 379 (+/- 373) microg x kg(-1) x d(-1) during both balances. Urinary excretion was high at 7 weeks of age, decreased to half at 12 week, and was negatively correlated (P < 0.01) with weight gain. Mean absorption was slightly positive at 7 weeks of age but zero or negative in most infants at 12 weeks of age. Retention was negative in all infants at both observation periods, except in one infant during the second balance. Clinical zinc deficiency developed in one infant at 12 weeks of age. CONCLUSIONS: Zinc balances in extremely low-birth-weight infants are highly variable and usually negative. Controlled trials are needed to assess need for and benefits and risks of zinc supplementation. PMID- 11396811 TI - Upper gastrointestinal mucosal disease in pediatric Crohn disease and ulcerative colitis: a blinded, controlled study. AB - BACKGROUND: Upper gastrointestinal endoscopic biopsies often show histologic abnormalities in Crohn disease. Consequently, it has been proposed that routine endoscopy could help to distinguish Crohn disease from ulcerative colitis. Surprisingly, however, recent case reports and an uncontrolled study suggested that similar abnormalities may occur in ulcerative colitis. Therefore, a blinded, controlled study was performed. METHODS: Esophageal, gastric antral, and duodenal biopsies from children with Crohn disease (n = 28) and ulcerative colitis (n = 14) were compared with those from controls undergoing endoscopy for suspected reflux esophagitis (n = 22). Two pathologists, unaware of patient identity and diagnosis, agreed on a consensus report. Severity of inflammation was scored semiquantitatively. Helicobacter pylori colonization was an exclusion criterion. RESULTS: Inflammation was reported as follows: esophagitis: controls 91%; Crohn disease: 72%; ulcerative colitis: 50%; gastritis: controls: 27%; Crohn disease: 92% (P < 0.001); ulcerative colitis: 69%; duodenitis: controls: 9%; Crohn disease: 33%; ulcerative colitis: 23%. In Crohn disease, granulomas were noted in 40% of patients (P = 0.001). Duodenal cryptitis was noted in 26% of patients with Crohn disease but not ulcerative colitis. In one patient with ulcerative colitis, neutrophilic infiltration of gastric glands was seen. Abnormalities seen in Crohn disease and ulcerative colitis included gastroduodenal ulceration (Crohn disease, 7%; ulcerative colitis, 8%), villus atrophy (Crohn disease, 11%; ulcerative colitis, 15%), and increased intraepithelial lymphocytes (Crohn disease, 15%; ulcerative colitis, 31% [P < 0.05]). None of these abnormalities was noted in the controls. CONCLUSION: Although the presence of granulomas can support a diagnosis of Crohn disease, severe inflammation and other abnormalities occur in the proximal gastrointestinal tract in Crohn disease and ulcerative colitis. PMID- 11396812 TI - Absence of Toll-like receptor 4 explains endotoxin hyporesponsiveness in human intestinal epithelium. AB - BACKGROUND: The Toll protein in Drosophila regulates dorsal ventral patterning during embryogenesis, and participates in antibacterial and antifungal host defense. Mammalian homologues are termed Toll-like receptors and, to date, nine have been cloned (TLRI-9) in humans. They are characterized by extracellular leucine-rich repeats and a cytoplasmic domain similar to the interleukin 1 receptor. Both TLR2 and TLR4 recognize various bacterial cell wall components including lipopolysaccharide (LPS). This results in the activation of the NFkappaB pathway. Peripheral blood mononuclear cells (PBMCs) express both TLR2 and TLR4. The authors hypothesized that the expression of TLR 2 and TLR4 in human intestinal epithelial cells differs from PBMCs because of the abundance of LPS in the intestinal lumen. METHODS: Epithelial cells were isolated from Caco-2 cells, fetal gut explants, and small bowel resection specimens using Hanks/ethylenediamine tetraacetic acid solution. PBMCs were used as positive controls. Ribonucleic acid (RNA) was isolated using the TRIzol method. Standard reverse transcription-polymerase chain reaction examined TLR2 and TLR4 messenger RNA (mRNA) expression. NFkappaB expression was determined using a luciferase reporter assay. RESULTS: TLR2 mRNA was highly expressed in PBMCs and was present in all human intestinal epithelial cells. TLR4 mRNA was detected only in PBMCs. TLR4 is not present in epithelium from children with inflammatory bowel disease. In Caco-2 cells, significant NFkappaB activation in response to LPS occurred only in the presence of TLR4 introduced by complementary deoxyribonucleic acid transfection. CONCLUSION: Absence of TLR4 is associated with endotoxin hyporesponsiveness of intestinal epithelial cells. TLR4 is not directly involved in inflammation of the intestinal epithelium. Although TLR2 is normally present in the epithelial cell, it plays a limited role in inflammation. It may be activated during conditions in which bacterial cell wall concentrations within the intestine are pathologically high. PMID- 11396813 TI - Saccharomyces boulardii does not stimulate mucosal hyperplasia after intestinal resection in the rat. AB - BACKGROUND: Polyamines have been shown to be important regulators of the intestinal adaptation process after massive bowel resection. Saccharomyces boulardii is yeast that has the ability to synthesize polyamines. Therefore. S. boulardii may be useful in the treatment of short bowel syndrome. METHODS: Twenty 150-g male Sprague-Dawley rats were subjected to 80% jejunoileal resection. Another 20 animals received transection and closure and served as pair fed controls. One half of the resected rats and one half of the controls were given S. boulardii 25 mg/day. RESULTS: After 2 weeks, mucosal mass (mg/cm bowel) did not differ between treated and non-treated animals despite the presence of a marked resection effect. Mucosal DNA, protein, and sucrase activity likewise did not differ. Subsequently, the experiment was repeated four times the original dose (100 mg/day) and found comparable results. In the proximal bowel, mucosal mass was 92+/-6 mg/cm in treated animals versus 107+/-8 mg/cm in untreated rats. In the distal small bowel, comparable values were 85+/-5 mg/cm and 88+/-4 mg/cm. Again, mucosal DNA, protein, and sucrase activity levels paralleled these results. CONCLUSIONS: Although S. boulardii may stimulate polyamine synthesis, it does not seem to be helpful in augmenting gut adaptation in this animal model of short bowel syndrome. PMID- 11396814 TI - Does cisapride influence cardiac rhythm? Results of a United States multicenter, double-blind, placebo-controlled pediatric study. AB - BACKGROUND: Major concerns about serious cardiac side effects underlie the recent decision by the FDA and Janssen Pharmaceutica (Titusville, NJ) to make cisapride available only through a limited access program. Concerns have grown despite the fact that most instances of prolonged QTc and other ventricular arrhythmias occurred while the drug was used concomitantly with contraindicated drugs. This study sought to analyze electrocardiograms (ECGs) from a multicenter pediatric study and to identify abnormalities in QTc interval associated with cisapride use. METHODS: Children between 6 months and 4 years of age were enrolled if they manifested symptoms of gastroesophageal reflux not responding to medical therapy for at least 6 weeks. In 49 subjects, ECGs obtained before and after randomization to receive 0.2 mg/kg dose three times daily or placebo were reviewed independently and blindly by two pediatric cardiologists. Placebo and active drug groups were compared for QTc and for change in QTc from baseline values after 3 to 8 weeks of treatment. RESULTS: Mean QTc among patients taking the drug was 408+/-18 ms. None was higher than 450 ms. Change between baseline and subsequent QTc at 3 to 8 weeks of treatment was 2+/-20 ms. CONCLUSIONS: In our study group of children without underlying cardiac disease or electrolyte imbalance, cisapride was found to have no significant effect on cardiac electrical function compared with placebo. These results are consistent with the drug's record of exceedingly infrequent cardiac events. Because the availability of this prokinetic is threatened, its safety and the safety and efficacy of alternative treatment options (including surgery) should be studied further. PMID- 11396815 TI - The effect of antenatal vitamin A and beta-carotene supplementation on gut integrity of infants of HIV-infected South African women. AB - BACKGROUND: Vitamin A is important for protection against diarrhea, and supplements may benefit gut function of infants of HIV-infected mothers. METHODS: We studied 238 infants of HIV-infected South African women participating in a randomized, double-blind, placebo-controlled trial of vitamin A during pregnancy (1.5 mg retinyl palmitate and 30 mg beta-carotene daily) plus 60 mg retinyl palmitate at delivery. The placebo group received identical placebo capsules at the same times. When infants were 1, 6, and 14 weeks of age, lactulose/mannitol dual sugar intestinal permeability tests were performed. RESULTS: Maternal vitamin A supplementation did not significantly affect infant gut permeability in the group as a whole at any time. By multiple regression analysis, HIV infection of the infant by 14 weeks was significantly associated with increased gut permeability at both 6 and 14 weeks. After controlling for birth weight, gestational age, current weight, feeding mode and recent morbidity, there was a trend toward an interaction between vitamin A supplementation and HIV infection (P = 0.086) at 14 weeks. Vitamin A made no difference to gut permeability of uninfected infants (lactulose/mannitol ratio for vitamin A group: 0.11, 95% confidence interval [CI] 0.08, 0.15, n = 73 and for placebo group: 0.09, 95% CI 0.06, 0.12, n = 76), but largely prevented the increase in the ratio of HIV infected infants (vitamin A group: 0.17, 95% CI 0.13, 0.23, n = 23; placebo group: 0.50, 95% CI 0.37, 0.68, n = 20). The effects on the lactulose/mannitol ratio were related to changes in lactulose, not mannitol, excretion. Vitamin A supplementation was associated with significantly lower lactulose excretion at 1 and 14 weeks, suggesting the major effect of vitamin A was on maintaining the integrity of gut tight junctions. CONCLUSIONS: Vitamin A supplementation of HIV infected pregnant women may prevent the deterioration in gut integrity in the subgroup of their infants who themselves become infected. Improving vitamin A status of HIV-infected infants may decrease their gastrointestinal morbidity. PMID- 11396816 TI - Delayed gastric emptying and gastroesophageal reflux: a pathophysiologic relationship. AB - BACKGROUND: Delayed gastric emptying (DGE) is frequent in patients with gastroesophageal reflux (GER), but its pathophysiologic role has not yet been established. To identify a relationship between DGE and GER, we assessed whether DGE increases esophageal acid exposure and the related importance of possible mechanisms. METHODS: Thirty pediatric patients with pathological GER were divided according to gastric emptying scintigraphy into a DGE group (n = 14) and normal emptying group (n = 16). The esophageal pH-monitoring parameters of the two groups were compared with respect to the individual variation between postprandial and fasting periods. RESULTS: Patients with DGE had less total acid exposure than did those with normal emptying, but patients in both groups had a pathological fraction of time when pH was below 4 in both the postprandial (median: 18 vs. 27.6; P = 0.49) and fasting (8.5 vs. 23.9; P = 0.01) periods. Patients in the normal-emptying group had similar fraction of time when pH was below 4 in the postprandial and fasting periods. However, patients in the group with DGE had a fraction of time when pH was below 4 in the postprandial period that was almost double that presented in fasting period (postprandial to fasting ratio: 2.11:0.90; P = 0.002). The postprandial to fasting ratio for episodes per hour was similar in the two groups (1.81 vs. 1.79; P = 0.62). Patients with DGE had a significantly higher frequency of long episodes in the postprandial period than did those with normal emptying (62.5% vs. 38.2%; P = 0.04). The occurrence of the longest episode in the postprandial period was also significantly higher for patients with DGE (57.1% vs. 6.2%; P = 0.003). CONCLUSIONS: DGE seems to accentuate postprandial reflux by increasing the volume of refluxate per episode of reflux through an underlying incompetent lower esophageal sphincter. PMID- 11396817 TI - Higher serum eosinophil cationic protein levels in children with cow's milk allergy. AB - BACKGROUND: In the pathogenesis of cow's milk allergy, abnormal immunologically mediated reactions play a basic role. Eosinophil activation also participates in the development of several allergies. The purpose of this study was to characterize the degree of this activation by measuring the serum level of eosinophil cationic protein (sECP) and establishing whether it is a useful parameter in monitoring oral cow's milk allergy. METHODS: The sECP level of 35 patients with previously confirmed cow's milk allergy (mean age, 16 months) was evaluated using a fluoroimmunoassay before the cow's milk rechallenge test and at 2 hours and 24 hours after cow's milk challenge. RESULTS: Of the 35 children with previously confirmed cow's milk allergy, 10 had positive clinical reactions after the milk rechallenge test, whereas 25 children had no reaction. The median sECP level of all the patients before the challenge test was significantly higher (12.4 microg/L) than that of the control group (4.3 microg/L) (P < 0.05). Two hours after the challenge, the median sECP of all patients (9.4 microg/L) was lower than the starting values. The median sECP levels were higher in children with positive challenge test results at all time points. However, this difference was not statistically significant. CONCLUSIONS: The normalization of sECP level may indicate the cessation of the cow's milk allergy. Therefore, the measurement of sECP may be helpful in determining the optimal time in which to repeat the challenge test, when the result will more likely be negative. The significant decrease of the sECP level 2 hours after the beginning of milk challenge test may be explained by the fact that this protein is excreted into the intestinal lumen. PMID- 11396818 TI - Focal nodular hyperplasia in children. PMID- 11396819 TI - Hypertransaminasemia in two children with hyperthyroidism. PMID- 11396820 TI - Chylous ascites: a case of child abuse and an overview of a rare condition. PMID- 11396821 TI - Celiac, fatty liver, and pancreatic insufficiency. PMID- 11396822 TI - Evans syndrome related to hepatitis B virus infection: a case that responded only to lamivudine therapy. PMID- 11396823 TI - Balloon dilation of double strictures after corrosive esophagitis. PMID- 11396824 TI - Colonic leiomyoma in a 12-year-old patient with tuberous sclerosis: a case report and brief review. PMID- 11396825 TI - Hereditary hemochromatosis not associated with common HFE gene mutation in Japanese siblings. PMID- 11396826 TI - Helicobacter pylori infection in Turkish children with recurrent abdominal pain. PMID- 11396827 TI - Alpha-antitrypin deficiency-associated liver disease in Turkish children. PMID- 11396828 TI - Effect of administration of Lactobacillus casei shirota on sodium balance in an infant with short bowel syndrome. PMID- 11396829 TI - A reproductive phase-dependent effect of dietary L-tryptophan on pineal gland and gonad of a nocturnal bird, Indian spotted owlet Athene brama. AB - Unlike other temperate owls, Indian spotted owlet Athene brama possesses a well developed pineal gland that secrets moderate amount of hydroxy- (serotonin) and methoxy- (melatonin) indoles in circulation. However, in this study, we have reported the response of this endocrine gland to exogenous L-Tryptophan (precursor of the above indoles), and also its effect on gonads of this nocturnal bird. During breeding phase or pineal inactive phase (March), oral treatment of L Trp (0.5 mg/100 g Bwt/day) significantly increased the pineal gland wt and plasma melatonin (MEL) level, while decreased the gonadal wt and plasma sex steroids levels (estradiol and progesterone in female and testosterone in male). Interestingly, during reproductively quiescent phase or pineal active phase (August), similar amount of L-Trp significantly decreased the plasma MEL level, while increased the above sex steroid levels in plasma. Finally, the results show a clear reproductive phase-dependent inverse effect of L-Trp on pineal gland and gonads for both sexes of the spotted owlets, and suggest that the therapeutic use of this amino acid would be a great advantage for controlling the reproduction of these economically important birds. PMID- 11396831 TI - The effect of egg size on yolk utilization and growth of rainbow trout alevins (Oncorhynchus mykiss Walbaum). AB - Changes in body weight and yolk weight were studied in rainbow trout alevins derived from large eggs (diameter >4 mm) and small eggs (egg diameter <4 mm) from the same female. Maximal body weight was reached later, and was higher, for alevins derived from large than from small eggs. The yolk was absorbed more quickly by the alevins of small eggs. Percentage of body water increased in alevins during fasting up to a limit of 91%. Although the limit was the same for both groups of alevins, it was reached more quickly by alevins from small eggs. Relative water content of the yolk did not seem to be influenced by egg size. These results suggest that original egg size had both a quantitative and qualitative effect on the early development of alevins. PMID- 11396830 TI - Histophysiological changes of the testicular tissue due to busulphan administration in the wild Indian house rat (Rattus rattus). AB - The results of the present study indicate the antispermatogenic activity of Busulphan or Myleran (1,4-dimethane-sulphonoxy butane) on the testicular tissue of adult male Indian house rat, Rattus rattus. Single oral dose of Busulphan (10 mg/Kg body weight) was administered and its activity was noticed at 10, 40, 70 and 100 days of posttreated animals. Histological observation and quantitative histological study indicates no major alteration in the relative percentages of primary spermatocytes, spermatid and Sertoli cells at 10 days of posttreatment. But there was a gradual decrease in the seminiferous tubular diameter at 40 and 70 days of post treated groups. However, the Leydig and Sertoli cells morphology and number remained normal in all the treatment groups. At 40 days, the normal cellular associations in all the tubules were disrupted. The tubules constituted only spermatogonia, Sertoli cells and some zygotene spermatocytes. At 70 days, repopulation of Type A, Type B spermatogonia, resting and zygotene spermatocytes occurred at this stage. The tubules were still devoid of pachytene spermatocytes, spermatid and spermatozoa. At 100 days, active spermatogenesis was observed in majority of the tubules. The various types of germ cell population were regaining towards normalcy. Histochemical studies clearly revealed that due to busulphan administration there was no major alteration in the intensities of some key enzymes (i.e. delta5 3beta-HSDH and 17beta-HSDH) involved in the biosynthesis of steroid hormones. Only the acid phosphatase activity was slightly depressed within the 40th and 70th days of posttreatment. Sudanophilic lipid materials increased in the interstitium of all the busulphan post treated groups. The changes which were noticed due to busulphan treatment regained normalcy at 100 days of post treated animals. The mode of action of Busulphan on the testicular tissue of adult Indian house rat (Rattus rattus) has been pointed out and discussed. PMID- 11396832 TI - On the micro-space theory of osmosis osmofiltration, "active" water-flow in membrane-gaps. AB - The author revised the conclusion drawn from his former experiments and recognized a complex phenomenon that he named osmofiltration. This phenomenon occurs by way of the inhibition of the diffusion within a micro-gap bordering the experimental membrane in the model. It may have a significance as a basic biological phenomenon in similar biological membranous structures, and even, it may have a medical importance. The author discusses his working hypothesis about the micro-space theory of osmosis. PMID- 11396833 TI - Effects of light on in vitro tuberization of the potato cultivar Desiree and its relatives. AB - Effects of different combinations of short days and dark treatment and different light intensities during the short days on in vitro tuberization of three potato cultivars were examined. Tuberization in Desiree, Cleopatra and Gracia was induced by short-day-treatment after 4 weeks culture under long days. To preserve natural endogenous hormonal balance of in vitro plantlets growth regulators were not added to the medium. The shorter the duration of short-days period the higher light intensity necessary for earlier tuber initiation. There was also a synchronizing effect of the high light intensity on tuber initiation but it depended on photoperiod-treatments and genotypes. The higher the light intensity during induction period the less favourable the effect of light applied after the induction period. PMID- 11396834 TI - Production of lignolytic and feed-back type enzymes by Phlebia radiata on different media. AB - Low molecular-weight compounds, structurally related to lignin, increase the production of laccase, lignin peroxidase, manganese dependent peroxidase, and feed-back type enzymes such as glucose oxidase, cellobioso-quinone oxidoreductase, and glyoxal oxidase in the culture of the white rot fungus Phlebia radiata growing on different carbon sources. PMID- 11396835 TI - Stress conditions applied to the interpretation of translation machinery. AB - Gene expression is regulated at the critical steps: a regulatory event occurs at the step which has a critical effect and is responsible for the limiting rate. Enzyme activity can be regulated at several different levels: transcriptional, translational or post-translational. In this review we describe (and illustrate with experimental data) plant stress which induces regulatory mechanisms at the translational and post-translational levels. We found evidence for autorepression regulatory system of ferritin biosynthesis. Based on the knowledge of the molecular mechanism of regulation, we believe that ferritin protects the environment against heavy metal ions and supplements biological system(s) with iron. The quinolizidine alkaloids' (QA) biosynthesis is lysine decarboxylase (LDC)-dependent. The available pool of LDC limits the conversion of lysine to cadaverine. The amount of LDC depends on transcriptional and translational efficiency. However, in the light of the presented data, we have evidence for a post-translational regulatory system, i.e. the activation of LDC from low to high activity enzyme through the conversion from higher to lower molecular weight form. The plant protection system is very efficient. Understanding of the defence systems such as plant response to stress, should provide us with a possibility of applying this knowledge in practice and finding novel applications. PMID- 11396836 TI - Pineal organ-like organization of the retina in megachiroptean bats. AB - Phylogenetically originated from photoreceptive structures, the pineal organ adapts the organism to circadian and circannual light periodicity of the environment, while the retina develops to a light-based locator. Bats have a nocturnal life and an echolocator orientation presumably modifying the task of photoreception. Looking for morphological basis of the special functions, in the present work we compared the fine structure and immunocytochemistry of the retina and pineal organ in micro- and megacrochiroptean bats. We found that there is a high similarity between the retina and pineal organ in megachiropterans when compared to other species investigated so far. Besides of photoreceptor derived pinealocytes, the pineal organ of both micro- and megachiropterans contain intrapineal neurons and/or ganglionic cells as well as glial cells. Like spherules and pedicles of retinal photoreceptors, axon-type processes of pinealocytes form synaptic ribbon containig terminals. Similar to retinal photoreceptors and neurons, pinealocytes and pineal neurons contain immunoreactive glutamate and aspartate. In addition, excitatory amino acids accumulate in the pineal neurohormonal endings and might have a role in the hormonal (serotonin?) release of the organ. Concerning the structure of the retina the highest similarity to the organization of the pineal organ was found in the megachiroptean fruit eating bats Cynopterus sphinx and Rusettus niloticus. The retina of these species forms folds and crypts in its photoreceptor layer. This organization is similar to the folds of the pineal wall successively developed during evolution. Since a folded photoreceptor layer is not viable for a photolocator screen in decoding two-dimensional images, we suppose that this peculiar organization of the megachiropteran retina is connected to a "pineal like" photometer task of the eye needed by these species active at night. PMID- 11396837 TI - Evaluation of genotoxity and mutagenicity of DL-p-chlorophenylalanine, its methyl ester and some N-acyl derivatives. AB - DL-p-chlorophenylalanine (PCPA) and its derivatives were evaluated for genotoxic effects using Escherichia coli and Bacillus subtilis strains lacking various DNA repair mechanisms in spottest and in suspension test. The mutagenic activity of studied compounds was determined by the Ames test. Reverse mutation test was performed with Salmonella typhimurium strains TA98, TA100, TA1535 and TA1537 without S9 mix. 0.02 M nitrosomethylurea (NMU) standard mutagen was used as a positive control. The results showed that the parent nonessential amino acid PCPA had no detectable genotoxic and mutagenic activities in bacteria. The methyl ester of this amino acid and its N-phenylacetyl derivative possessed weak genotoxicity. Meanwhile N-sec-butyloxycarbonyl, N-benzyloxycarbonyl, N-(p nitrophenylacetyl) and N-(p-nitrophenoxyacetyl) derivatives of DL-p chlorophenylalanine exhibited appreciable genotoxicity. Among the seven tested compounds only N-benzyloxycarbonyl and N-(p-nitrophenoxyacetyl) derivatives of DL p-chlorophenylalanine have been found to be mutagenic. Only parent PCPA possessed antimutagenic properties in respect of nitrosomethylurea. The structural modification, which strongly affects genotoxicity and mutagenicity perhaps may be due to steric hydrance of the substituents, causing interference with enzyme and DNA interactions. PMID- 11396838 TI - Role of environmental factors on the reproducibility of Lemna test. AB - Lemna minor is a species easy to collect and culture in laboratory, and can give rapid test results. However, in order to standardise toxicity tests using Lemna minor as test organism, it is important to find out what natural variability different populations might have. Five Lemna populations were used for comparison. It contained two standard cultures and three populations collected in natural habitats. Potassium dichromate was applied as test material. Lemna populations cultured under the same conditions showed different T(D) and LC50 values. There is an inverse relation between the sensitivity and T(D) of the strains. It is supposed that growth rate and sensitivity of Lemna populations depend on environmental factors characterising the habitat in which the given popluation originally lives. PMID- 11396839 TI - Sexual dimorphism of glial fibrillary acidic protein (GFAP) immunoreactivity in the rat interpeduncular nucleus. AB - The intensity of immunostaining for the glial fibrillary acidic protein (GFAP) is outstandingly high in the interpeduncular nucleus. This nucleus was compared in males and females for its GFAP immunoreaction. Immunohistochemistry was carried out on free floating vibratome slices and evaluated by surface densitometry. While in males the reactions were similar, females showed individual variations. Since the interpeduncular nucleus is a hormonally inactive brain area where gonadal hormones do not induce plastic synaptic changes, it is concluded that concerning this astroglial marker a sexual dimorphism exists also outside the "endocrine brain". PMID- 11396840 TI - Hypoglycemia activates compensatory mechanism of glucose metabolism of brain. AB - The effect of plasma glucose concentration on the cerebral uptake of [18F] fluorodeoxy-D-glucose (FDG) was studied in a broad concentration range in a rabbit brain model using dynamic FDG PET measurements. Hypoglycemic and hyperglycemic conditions were maintained by manipulating plasma glucose applying i.v. glucose or insulin load. FDG utilization (K) and cerebral glucose metabolic rate (CGMR) were evaluated in a plasma glucose concentration range between 0.5 mM and 26 mM from the kinetic constant k1, k2, k3 obtained by the Sokoloff model of FDG accumulation. A decreasing set of standard FDG uptake values found with increasing blood glucose concentration was explained by competition between the plasma glucose and the radiopharmacon FDG. A similar trend was observed for the forward kinetic constants k1, and k3 in the entire concentration range studied. The same decreasing tendency of k2 was of a smaller magnitude and was reverted at the lowest glucose concentrations where a pronounced decrease of this backward transport rate constant was detected. Our kinetic data indicate a modulation of the kinetics of carbohydrate metabolism by the blood glucose concentration and report on a special mechanism compensating for the low glucose supply under conditions of extremely low blood glucose level. PMID- 11396841 TI - A slow outward current and a hypoosmolality induced anion conductance in embryonic chicken osteoclasts. AB - In this paper we report on a hypoosmolality induced current, I(osmo), in embryonic chicken osteoclasts, which could only be studied when blocking a simultaneously active, unidentified slow outward current, I(slo). I(slo) was observed in all of the examined cells when both the intracellular and extracellular solutions contained sodium as the major cation and no potassium. The current was outwardly rectifying and activated at membrane potentials more positive than -44 +/- 12 mV (n = 31). The time to half activation of the current was also voltage dependent and was 350 ms at Vm = +80 mV, and 78 ms at Vm = +120 mV. The current did not inactivate during periods up to 5 s. Extracellular 4-AP (5 mM), TEA (5 mM) and Ba2+ (1 mM), blockers of K+ conductances in chicken osteoclasts, did not influence I(slo). However, I(slo) was inhibited by 50 microM extracellular verapamil, which allowed us to study I(osmo) in isolation. Exposure of the osteoclasts to hypotonic solution resulted in the development of a depolarization activated I(osmo). It developed after a 1-min delay and reached its maximum within 10 minutes. Half-maximal activation occurred after 4.4 +/- 0.9 min (n = 9). The current activated within a few ms upon depolarization and did not inactivate during at least 5 sec. I(osmo) reversed around the calculated Nernst potential for Cl- (E(Cl) = +7.3 mV and V(rev) = +5.4 +/- 3.6 mV, n = 9). The underlying conductance, G(osmo) exhibited moderate outward rectification around 0 mV in symmetrical Cl- solutions. Ion substitution experiments showed that G(osmo) is an anion conductance with P(Cl) approximately = P(F) > P(gluc) >> P(Na). I(osmo) was blocked by 0.5 mM SITS but 50 microM verapamil, 5 mM TEA, 5 mM 4-AP, 1 mM Ba2+, 50 microM cytochalasin D and 0.5 mM alendronate did not have any effect on the current. Cl- currents have been implicated in charge neutralization during osteoclastic acid secretion for bone resorption. The present results imply that osmolality may be a factor controlling this charge neutralization. PMID- 11396842 TI - Effects of endogenous and exogenous glucocorticoids on liver differentiation. AB - The effects of maternal bilateral adrenalectomy on day 1 of gestation and betamethasone treatment on fetal liver development were compared, in terms of biochemical and morphological parameters. For fetuses 20 days old (E20), absence of maternal glucocorticoids during gestation caused an increase in the number of nuclei in whole livers, and a significantly decrease of both body weight and protein content per nucleus, in comparison with the control group (C). Betamethasone injection on days 15, 16 and 17 of gestation into adrenalectomized pregnant rats (ADX + BET) did not completely prevent these effects. The electron microscopic analysis of the ADX fetal liver (E20) showed some hepatocyte lesions such as loss of cytoplasmic organelles, increase in hematopoietic cell number as well as a lower cellular maturation in comparison with the control group. The fetal liver from ADX + BET mothers 20 days after gestation displayed a noticeable involution of the hematopoietic component in spite of its relatively immature stage. However, there was no significant change in the degree of fetal hepatocyte lesions. Therefore, supply of maternal glucocorticoids from the beginning of gestation is essential for maintenance of the integral structure of the rat fetal hepatic parenchyma, for the correct maturation of the blood strains and for the beginning of involution of the hematopoietic tissue at the end of gestation. PMID- 11396843 TI - Gut morphology and morphometry in the epauletted Wahlberg's fruit bat (Epomophorus wahlbergi, Sundevall, 1846). AB - The morphological adaptations of the fruit bat small intestine to which the high functional efficiency could be related and the possible landmarks delineating the various parts of the gut were examined. The stomach was the carnivorous type with large rugae spanning the entire luminal aspect down to the pyloric sphincter, which was reflected internally as a prominent fold. Externally, the intestine was a continuous tube uninterrupted by any structures. The cranial fifth of the small gut had long, branching and anastomosing villi, which caudally turned to finger like discrete structures that became rather short and stumpy and diminished at the beginning of the colon. The colon had longitudinal folds that were macroscopically discernible from the mucosal aspect of the opened intestine and that continued into the rectum. The small gut formed 94% of the whole intestinal length, the colon and the rectum taking 4 and 2%, respectively. Ultrastructurally, the enterocyte showed a prominent brush border and the lateral membranes were modified into numerous tortuous interdigitating processes. Adjacent enterocytes were joined by these processes through desmosomes. The processes also participated in pinocytotic fluid uptake from the intercellular spaces with resultant numerous intracellular vacuoles of varied sizes. Solutes absorbed into the cells were probably first passed into the intercellular compartment to create a concentration gradient thus enhancing further absorption into the cell. We conclude that the uniquely elaborate ultrastructure of the enteric epithelium coupled with the vast microvillous surface areas reported elsewhere are partly responsible for the very high absorption rates reported in the fruit bat small intestine. PMID- 11396844 TI - Reproductive phase depedent circadian variation in the pineal biochemical constituents of Indian palm squirrel, Funambulus pennanti. AB - In mammals, pineal gland is intimately concerned with the co-ordination of rhythm physiology. Biochemical characteristics of pineal gland in man and other mammals may provide strong, yet sometimes elusive support for the belief in functional individuality and probable importance of this tiny gland. In seasonal breeding animals, pineal gland function is very much dependent on the reproductive status. Therefore, the aim of this experiment is to note the circadian rhythmicity of different biochemical constituents of pineal gland during active and inactive phases of reproductive cycle of a seasonally breeding rodent, F. pennanti. In the present study, pineal biochemical constituents i.e. protein and cholesterol showed higher values during daytime (1400 h). The plasma melatonin level presented two peaks during active (April; at 1800 h and 0200 h) and inactive (December; at 1400 h and 0200 h) phases of reproductive cycle. The pineal protein, cholesterol and plasma melatonin values in term of basal and peak levels were higher during the reproductive inactive/pineal active phase. Therefore, pineal--also known to have antigonadotropic properties and cholesterol which appears conjugated with pineal serotonin, presented circadian rhythmicity along with the plasma level of melatonin. This rhythmicity noted in present study was dependent on the reproductive and pineal activity status, and might be regulated by the sex steroid receptor present on the pineal gland. PMID- 11396845 TI - Castration-induced hyperactivity of seminal vesicle in the catfish Clarias batrachus: a case of paradox and blockade by antiandrogen (cyproterone acetate) treatment. AB - Castration of the catfish Clarias batrachus in late preparatory-early prespawning phase (April-May) caused time-dependent stimulatory effect on morphology, weight, and in the concentrations of biochemical correlates, such as total proteins, fructose, hexosamines and sialic acid in the seminal vesicle (SV). The peak changes were noticed on week 4 of castration. The hyperactivity was related to augmented production of testosterone by the SV of castrates with the levels significantly high from week 3 onwards. As a result, serum testosterone level fluctuated with a significant decrease in the first and fifth weeks, a significant increase in the third week, and no significant difference in the second and fourth weeks. Serum E2 level decreased significantly throughout. Cyproterone acetate treatment (CA; 1 mg/fish daily for 21 days) from the second day of castration decreased the size and weight of the SV and the concentrations of total proteins, hexosamines, fructose and sialic acid. The antiandrogen treatment did not alter serum testosterone level but the E2 level was significantly decreased. It is concluded that the hypersecretory activity of the SV in castrates is a sequel to local synthesis and action of testosterone and the effect could be prevented by CA by blocking androgen actions. PMID- 11396846 TI - Scale invariance in biology: coincidence or footprint of a universal mechanism? AB - In this article, we present a self-contained review of recent work on complex biological systems which exhibit no characteristic scale. This property can manifest itself with fractals (spatial scale invariance), flicker noise or 1/f noise where f denotes the frequency of a signal (temporal scale invariance) and power laws (scale invariance in the size and duration of events in the dynamics of the system). A hypothesis recently put forward to explain these scale-free phenomomena is criticality, a notion introduced by physicists while studying phase transitions in materials, where systems spontaneously arrange themselves in an unstable manner similar, for instance, to a row of dominoes. Here, we review in a critical manner work which investigates to what extent this idea can be generalized to biology. More precisely, we start with a brief introduction to the concepts of absence of characteristic scale (power-law distributions, fractals and 1/f-noise) and of critical phenomena. We then review typical mathematical models exhibiting such properties: edge of chaos, cellular automata and self organized critical models. These notions are then brought together to see to what extent they can account for the scale invariance observed in ecology, evolution of species, type III epidemics and some aspects of the central nervous system. This article also discusses how the notion of scale invariance can give important insights into the workings of biological systems. PMID- 11396847 TI - Individual versus social complexity, with particular reference to ant colonies. AB - Insect societies colonies of ants, bees, wasps and termites--vary enormously in their social complexity. Social complexity is a broadly used term that encompasses many individual and colony-level traits and characteristics such as colony size, polymorphism and foraging strategy. A number of earlier studies have considered the relationships among various correlates of social complexity in insect societies; in this review, we build upon those studies by proposing additional correlates and show how all correlates can be integrated in a common explanatory framework. The various correlates are divided among four broad categories (sections). Under 'polyphenism' we consider the differences among individuals, in particular focusing upon 'caste' and specialization of individuals. This is followed by a section on 'totipotency' in which we consider the autonomy and subjugation of individuals. Under this heading we consider various aspects such as intracolony conflict, worker reproductive potential and physiological or morphological restrictions which limit individuals' capacities to perform a range of tasks or functions. A section entitled 'organization of work' considers a variety of aspects, e.g. the ability to tackle group, team or partitioned tasks, foraging strategies and colony reliability and efficiency. A final section, 'communication and functional integration', considers how individual activity is coordinated to produce an integrated and adaptive colony. Within each section we use illustrative examples drawn from the social insect literature (mostly from ants, for which there is the best data) to illustrate concepts or trends and make a number of predictions concerning how a particular trait is expected to correlate with other aspects of social complexity. Within each section we also expand the scope of the arguments to consider these relationships in a much broader sense of'sociality' by drawing parallels with other 'social' entities such as multicellular individuals, which can be understood as 'societies' of cells. The aim is to draw out any parallels and common causal relationships among the correlates. Two themes run through the study. The first is the role of colony size as an important factor affecting social complexity. The second is the complexity of individual workers in relation to the complexity of the colony. Consequently, this is an ideal opportunity to test a previously proposed hypothesis that 'individuals of highly social ant species are less complex than individuals from simple ant species' in light of numerous social correlates. Our findings support this hypothesis. In summary, we conclude that, in general, complex societies are characterized by large colony size, worker polymorphism, strong behavioural specialization and loss of totipotency in its workers, low individual complexity, decentralized colony control and high system redundancy, low individual competence, a high degree of worker cooperation wher tackling tasks, group foraging strategies, high tempo, multi-chambered tailor-made nests, high functional integration, relatively greater use of cues and modulatory signals to coordinate individuals and heterogeneous patterns of worker-worker interaction. PMID- 11396848 TI - On the origin and evolution of the human immunodeficiency virus (HIV). AB - The human AIDS viruses--HIV-1 and HIV-2--impose major burdens on the health and economic status of many developing countries. Surveys of other animal species have revealed that related viruses--the SIVs are widespread in a large number of African simian primates where they do not appear to cause disease. Phylogenetic analyses indicate that these SIVs are the reservoirs for the human viruses, with SIVsm from the sooty mangabey monkey the most likely source of HIV-2, and SIVcpz from the common chimpanzee the progenitor population for HIV-1. Although it is clear that AIDS has a zoonotic origin, it is less certain when HIV-1 and HIV-2 first entered human populations and whether cross-species viral transmission is common among primates. Within infected individuals the process of HIV evolution takes the form of an arms race, with the virus continually fixing mutations by natural selection which allow it to escape from host immune responses. The arms race is less intense in SIV-infected monkeys, where a weaker immune response generates less selective pressure on the virus. Such a difference in virus-host interaction, along with a broadening of co-receptor usage such that HIV strains are able to infect cells with both CCR5 and CXCR4 chemokine receptors, may explain the increased virulence of HIV in humans compared to SIV in other primates. PMID- 11396849 TI - Are ecological and evolutionary theories scientific? AB - Scientists observe nature, search for generalizations, and provide explanations for why the world is as it is. Generalizations are of two kinds. The first are descriptive and inductive, such as Boyle's Law. They are derived from observations and therefore refer to observables (in this case, pressure and volume). The second are often imaginative and form the axioms of a deductive theory, such as Newton's Laws of Motion. They often refer to unobservables (e.g. inertia and gravitation). Biology has many inductive generalizations (e.g. Bergmann's Rule and 'all cells arise from preexisting cells') but few, if any, recognized universal laws and virtually no deductive theory. Many biologists and philosophers of biology have agreed that predictive theory is inappropriate in biology, which is said to be more complex than physics, and that one can have nonpredictive explanations, such as the neo-Darwinian Theory of Evolution by Natural Selection. Other philosophers dismiss nonpredictive, explanatory theories, including evolutionary 'theory', as metaphysics. Most biologists do not think of themselves as philosophers or give much thought to the philosophical basis of their research. Nevertheless, their philosophy shows in the way they do research. The plethora of ad hoc (i.e. not universal) hypotheses indicates that biologists are reluctant inductivists in that the search for generalization does not have a high priority. Biologists test their hypotheses by verification. Theoretical physicists, in contrast, are deductive unifiers and test their explanatory hypotheses by falsification. I argue that theoretical biology (concerned with unobservables, such as fitness and natural selection) is not scientific because it lacks universal laws and predictive theory. In order to make this argument, I review the differences between verificationism and falsificationism, induction and deduction, and descriptive and explanatory laws. I show how these differ with a specific example of a successful and still useful (even if now superseded as explanatory) deductive theory, Newton's Theory of Motion. I also review some of the philosophical views expressed on these topics because philosophers seem to be even more divided than biologists, which is not at all helpful. The fact that biology does not have predictive theories does not constitute irrefutable evidence that it cannot have them. The only way to falsify this philosophical hypothesis, however, is to produce a predictive theory with universal biological laws. I have proposed such a theory, but it has been presented piecemeal. At the end of this paper, I bring the pieces together into a deductive theory on the evolution of life history traits (e.g. clutch size, mating relationships, sexual size dimorphism). PMID- 11396850 TI - Curly tail: a 50-year history of the mouse spina bifida model. AB - This paper reviews 50 years of progress towards understanding the aetiology and pathogenesis of neural tube defects (NTD) in the curly tail (ct) mutant mouse. More than 45 papers have been published on various aspects of curly tail with the result that it is now the best understood mouse model of NTD pathogenesis. The failure of closure of the spinal neural tube, which leads to spina bifida in this mouse, has been traced back to a tissue-specific defect of cell proliferation in the tail bud of the E9.5 embryo. This cell proliferation defect results in a growth imbalance in the caudal region that generates ventral curvature of the body axis. Neurulation movements are opposed, leading to delayed neuropore closure and spina bifida, or tail defects. It is interesting to reflect that these advances have been achieved in the absence of information on the nature of the ct gene product, which remains unidentified. In addition to the principal ct gene, which maps to distal Chromosome 4, the curly tail phenotype is influenced by several modifier genes and by environmental factors. NTD in curly tail are resistant to folic acid, as is thought to be the case in 30% of human NTD, whereas they can be prevented by myo-inositol. These and other features of NTD in this system bear striking similarities to the situation in humans, making curly tail a model for understanding a sub-type folic acid-resistant human NTD. PMID- 11396851 TI - A comparative study of an olfactory epithelial area lacking olfactory neurons and a nearby presence of TGF-alpha-like immunoreactive olfactory neurons. AB - Our previous study has shown that ddY mice have special patches of nasal epithelium in the posterior roof of the nasal cavity that exclusively consists of olfactory supporting cells and horizontal basal cells. Here, we extend this finding to Balb/c and DBA/2 mice, Wistar and Sprague-Dawley rats, hamsters, and guinea pigs. In the mice, rats, and hamsters studied, the patches lacked olfactory cells and their precursor, globose basal cells. In rats and hamsters, the supporting cells were arranged in a single layer, in mice as three or four layers. Horizontal basal cells were located in a single layer in these species. In the guinea pigs, the specialized roof structure was less clear and could be seen at the level of ultrastructure as an olfactory neuron-lacking area. Distinct populations of transforming growth factor (TGF)-alpha-like immunoreactive olfactory cells occupied an area close to the epithelial patches. In this region, the TGF-alpha-like immunoreactive neurons were negative for the usual olfactory markers, either OMP or protein gene product (PGP) 9.5 or beta-tubulin. These cells are suggested to project to the so-called 'necklace glomeruli' and use a different cGMP-driven, transduction pathway. Three-dimensional analysis of double labeled (TGF-alpha, PGP9.5) serial sections revealed a unique relation among the epithelial patches, TGF-alpha-like immunoreactive neurons and olfactory epithelium. PMID- 11396852 TI - Proliferation and differentiation of thyrotrophs in the pars distalis of the rat pituitary gland during the fetal and postnatal period. AB - Pituitary glands from rat fetuses (gestational age 17.5-21.5 days) and rat pups (3, 7, 10, 14, 28 days old) were labeled with bromodeoxyuridine (BrdU) 2 h prior to sacrifice and embedded in paraffin. Sections were consecutively immunostained with anti-BrdU and anti-rat TSH. The number of cells stained with anti-BrdU, anti rTSH, or both of them were counted. The area of the section and the volume of the pituitary were measured and the number of immunostained cells per mm3 or per pituitary was calculated. Thyrotrophs were few in 17.5 day-fetuses but increased thereafter, with a rapid increase during the 2nd week after birth. The number of cells labeled with both BrdU and TSH peaked at 7 days after birth. It was estimated that about 1/5 of the thyrotrophs increased during this period was derived from the mitosis of existing thyrotrophs. PMID- 11396853 TI - Ablation of paternal accessory sex glands is detrimental to embryo development during implantation. AB - The accessory sex glands are present in most mammals, but their function(s) have not yet been clearly defined. In the golden hamster, removal of all the glands or the ventral prostate alone have been shown to considerably reduce fertility, while the effect is milder if the ampullary glands only are removed. In this study, embryo development from the 5th to the 7th day after mating are examined. Structural and morphometric criteria such as cell number, cell density, embryo volume, volume fraction of proamniotic cavity further revealed that abnormalities can be demonstrated as early as day 5 in the embryos sired by males with the ventral prostate gland alone or all glands ablated. Twin implantation and deviation from normal implanted axis are also observed. This is likely to be attributed to attenuated cell proliferation, as indicated by proliferating cell antigen labelling and more necrotic cell death. Taken together, exposure of sperm to secretions of the male accessory sex glands in particular, the ventral prostate, is important for differentiation and multiplication of cells after the embryo has implanted. PMID- 11396854 TI - Development of the cyto- and chemoarchitectural organization of the rat nucleus of the solitary tract. AB - The nucleus of the solitary tract (NST) is the major visceral sensory nucleus in the brainstem. The development of the rat nucleus of the solitary tract was followed during late prenatal and early postnatal life in order to determine when subnuclear organization and chemoarchitectural features develop. In Nissl-stained sections, the nucleus of the solitary tract becomes visible as a distinct cluster of cells by about E17. Between E17 and E19, a profound change in the Nissl stained appearance of the nucleus occurred, so that by E19 all the subnuclei were discernible. Acetylcholinesterase activity in the developing NST showed an early period of rapid differentiation (E15 to E17), while by E19 the basic adult pattern of distribution of this enzyme had already been achieved. The subnuclei of the NST began to show clear differential staining for nicotinamide adenine dinucleotide phosphate diaphorase at about the same time as reactivity for that enzyme first appeared (E19). With respect to calbindin- and calretinin immunoreactive neurons within the nucleus, many of the chemoarchitectural features associated with these two markers were obvious even by late fetal life. For example, in the central subnucleus, a strongly labelled, dense population of calbindin-immunoreactive neurons was present from E17; while in calretinin immunoreacted material, this subnucleus was prominent because of its immunonegativity also from E17. Nevertheless, the total number of calbindin- and calretinin-immunoreactive neurons in the NST did not peak until late postnatal life. Tyrosine hydroxylase immunoreactive neurons were visible from E15, began differentiation by E17 and were distributed in a similar pattern to the adult from E19. Substance P immunoreactivity in the NST was also very similar to the adult pattern by E19. Many of these immunochemical and histochemical markers indicate a similar pattern of development, i.e. a rapid period of differentiation until E19, by which time a relatively stable adult-like pattern has been attained. The present findings indicate that many of the cyto- and chemoarchitectural features of this nucleus are present well before birth, by which time the nucleus must serve vitally important functions such as relaying information for control of respiration and the circulation. PMID- 11396855 TI - Enrichment of unipolar brush cell-like neurons in primary rat cerebellar cultures. AB - Unipolar brush cells are a distinct class of excitatory interneurons situated in the granular layer of the cerebellar cortex, where they form giant synapses with individual mossy fiber terminals. We have previously shown that primary cerebellar cell cultures from embryonic and postnatal rodents contain neurons displaying morphological and chemical phenotypes characteristic of unipolar brush cells in situ, including intense staining with calretinin antiserum. In cultures from both embryonic and postnatal rats, the large majority of calretinin-positive neurons are unipolar brush cells, while granule cells are usually calretinin negative. A small percentage of putative Golgi/Lugaro cells also express calretinin. We demonstrate here that the developmental stage of the source tissue, the concentration of potassium in the medium, and treatment with glutamate after differentiation have substantial effects on the density of putative unipolar brush cells in the cultures. In dissociated cultures obtained from embryos at gestation day E18 and E20 and from pups at postnatal day P0, P2, P5, P8, and P10 grown in 25 mM KCl, the percentage calretinin-positive cells progressively decreases from 24% to 0.1% of total cells. In cultures from E20 embryos grown in physiological potassium (5 mM KCl), calretinin-positive cells are enriched to approximately 60% of total cells, while the majority of calretinin-negative cells die. In embryonic cultures exposed to high concentrations of glutamate after 12 days in vitro, calretinin-positive neurons have a survival advantage over calretinin-negative cells and represent up to 83% of total cells. PMID- 11396856 TI - Morphogenesis of the bovine rete testis: extratesticular rete, mesonephros and establishment of the definitive urogenital junction. AB - The development of the extratesticular rete, the regression of the mesonephros and the establishment of the urogenital junction between rete testis and efferent ductules were investigated in 67 bovine embryos and fetuses collected in the period from day 29 through day 250 post conception. The results were obtained by immunohistochemistry and by the study of semithin sections. At about day 30, the large mesonephros contains a peculiar Malpighian body in its cranial part, generally referred to as the mesonephric giant corpuscle, which is connected to the Wolffian duct by a series of well-developed and functioning mesonephric tubules. This set of primary mesonephric tubules, however, will not participate in the formation of the definitive urogenital junction, but will regress and soon disappear completely. The efferent ductules in the bovine are represented by another set of secondary mesonephric tubules that grow out from the dorsal aspect of the mesonephric giant corpuscle at about day 50. Transiently, the lumina of the sprouting efferent ductules are plugged by invading intraductular blood vessels, probably representing rudimentary glomeruli. The proximal portions of the newly-formed efferent ductules establish side-to-end contacts with extensions of the extratesticular rete that has bypassed the regressing giant corpuscle. At 85 days, the efferent ductules have reached the Wolffian duct and open into it. At 150 days, the channels of the extratesticular rete display a patent lumen and now form end-to-end anastomoses with the efferent ductules. The proliferating mesenchymal cells surrounding the epithelia of the efferent ductules have arranged in several concentric layers at about 85 days. These mesenchymal cells are the precursors of the periductular musculature and are reached by the first nerve fibers at about day 130. PMID- 11396857 TI - Detailed organization of nitric oxide synthase, vasopressin and oxytocin immunoreactive cell bodies in the supraoptic nucleus of the female rat. AB - The anatomical distribution and quantitative relations of cell bodies containing neuronal nitric oxide synthase (nNOS), 8-arginine vasopressin (AVP) and oxytocin (OT) were examined throughout the supraoptic nucleus (SON) of the female rat by means of immunocytochemical and NADPH-diaphorase (NADPH-d) histochemical techniques using a triple labelling methodology. Seven chemically defined populations of neurons containing all combinations of either nNOS, AVP or OT were identified. nNOS-containing (NADPH-d positive) neurons, amounting to about 40% of all neurons counted, were most frequent in central and dorsal regions, and were evenly distributed along the rostro-caudal axis. Two small nNOS-positive neuronal populations were preferentially located dorso-centrally in the nucleus: nNOS positive neurons containing both AVP- and OT-immunoreactivity, and neurons only containing nNOS. Slightly less than half of all nNOS-positive neurons contained AVP, and a similar share of nNOS-positive neurons contained OT. The occurrence of nNOS-positive/AVP-containing neurons was highest in the caudal half, whereas that of nNOS-positive/OT-neurons was highest in the rostral half of SON. The data demonstrate new findings concerning the anatomical organization and co localization patterns of nNOS-, AVP- and OT-containing neuronal populations in SON. We conclude that the absolute and relative occurrence of the identified neuronal populations vary markedly in different parts of SON. This is important to take into consideration when performing, and evaluating experimental investigations concerned with neurochemical changes in SON. PMID- 11396858 TI - Mechanisms of protection induced by live attenuated simian immunodeficiency virus: III. Viral interference and the role of CD8+ T-cells and beta-chemokines in the inhibition of virus infection of PBMCs in vitro. AB - In this study, we investigated whether a type of retroviral interference might be one mechanism that mediates the powerful protection induced by live attenuated SIVC8. Our results show that retroviral interference could be demonstrated between SIV and SHIV-HXBc2 in human T-cell lines chronically infected with either SIVC8 or SIVJ5. Lymphocytes from macaques infected with live attenuated SIVC8 were significantly less sensitive (P < 0.05) to in vitro infection by virulent SIVJ5 and SHIV-HXBc2 than were lymphocytes from naive controls. However, this significant difference in the sensitivity of lymphocytes to virus infection was not observed for more efficiently replicating viruses such as SHIVSF33 and SIVsm3. Virus growth was significantly enhanced (P < 0.01) by depletion of CD8+ T cells, suggesting a role for these cells in the control of SIV replication, both in vitro and in vivo. We found that levels of the beta-chemokines regulated upon activation, normal T-cell expressed and secreted, macrophage inflammatory protein 1alpha and macrophage inflammatory protein-1beta did not correlate with inhibition of virus replication. Taken together, our findings do not support the hypothesis that retroviral interference is the mechanism by which live attenuated SIVC8 induces protection. PMID- 11396859 TI - Prevalence of hepatitis B virus infections in nonhuman primates. AB - The aim of this study was to determine the prevalence of hepatitis B virus (HBV) infection in nonhuman primates. Serum samples from Europe, Thailand and Vietnam were analyzed. Sera obtained from 262 apes and 454 monkeys were tested for HBV infection serologically and for HBV DNA using nested PCR (nPCR). A total number of 198 ape sera and all but one (Cercopithecus aethiops) of the 4543 monkey sera had no serological signs of HBV infection. Among the 64 of 262 (24.4%) seropositive ape sera, we found, as in humans, different stages of HBV infection: very early HBV infection, active infection with high level of infectivity, virus carriers with low infectivity, and passed HBV infection. In the cases with passed infection, 47.8% harbored HBV DNA in the presence of protective antibodies to the HBV surface antigen (HBsAb). This indicates HBV persistence in apes despite immune control. In contrast to apes, in monkeys HBV infection is a very rare event. PMID- 11396860 TI - A prevalence survey for zoonotic enteric bacteria in a research monkey colony with specific emphasis on the occurrence of enteric Yersinia. AB - Transmissible pathogenic and opportunistic zoonotic enteric bacteria comprise a recognized occupational health threat to exposed humans from non-human primates (NHPs). In an effort to evaluate the occurrence of selected enteric organisms with zoonotic and biohazard potential in a research colony setting, we performed a prevalence study examining 61 juvenile and young adult rhesus macaques participating in a transplant immunology project. Primary emphasis was directed specifically to detection of pathogenic enteric Yersinia, less well-documented and reported NHP pathogens possessing recognized significant human disease potential. NHPs were surveyed by rectal culture during routine health monitoring on three separate occasions, and samples incubated using appropriate media and specific selective culture methods. Enteric organisms potentially transmissible to humans were subcultured and identified to genus and species. Significant human pathogens of the Salmonella/Shigella, Campylobacter, and enteric Yersinia groups were not isolated throughout the survey, suggesting prevalence of these organisms may generally be quite low. PMID- 11396861 TI - Comparative analysis of natural killer cell activity, lymphoproliferation and lymphocyte surface antigen expression in nonhuman primates housed at the CIRMF Primate Center, Gabon. AB - Six different species of nonhuman primates housed at the CIRMF Primate Center, cynomolgus monkeys (Macaca fascicularis), rhesus monkeys (Macaca mulatta), mandrills (Mandrillus sphinx), vervets (Cercopithecus aethiops pygerythrus), chimpanzees (Pan troglodyte) and baboons (Papio hamadryas), were evaluated for their natural killer cell activity and for the ability of their peripheral blood mononuclear cells to proliferate in response to known mitogens (concanavalin A, phytohemagglutinin and staphylococcal enterotoxin A) and to react with a panel of mouse monoclonal antibodies directed against human leukocyte surface antigens. Basic information on normal immune functions in these primates is important because of their use as experimental animal models for the study of human diseases such as acquired immunodeficiency syndrome (AIDS), hepatitis, loiasis and malaria. PMID- 11396862 TI - Identification of primitive hematopoietic progenitor cells in the rhesus macaque. AB - The close phylogenetic relationship of macaques to humans has resulted in their widespread use as a preclinical model for bone marrow transplantation and stem cell gene therapy. To facilitate further use of this model, we undertook analysis of hematopoietic cells using multiparametric flow cytometric analysis. Rhesus CD34+CD38- cells displayed a number of characteristics of primitive hematopoietic cells, including low forward and orthogonal scatter and the lack of expression of lineage-specific markers or human lymphocyte antigen-DR. Four-color flow cytometric analysis demonstrated that rhesus CD34+CD38- cells were heterogenous with respect to Thy-1 expression and were CD59dim. Quantitative limiting dilution long-term culture-initiating cell (LTC-IC) analysis demonstrated that CD34+CD38- cells were approximately 150-fold enriched for LTC-IC as compared with unfractionated bone marrow, and occurred at a frequency similar to that previously reported in humans. Thus, as in humans, the CD34+38- population of rhesus macaque bone marrow is enriched for primitive, multipotent hematopoietic progenitor cells. PMID- 11396863 TI - Ultrasonographic findings of functioning renal allografts in the cynomolgus monkey (Macaca fascicularis). AB - PURPOSE: The purpose of this study was to describe the findings of serial ultrasound investigations of functioning and histologically normal renal allografts in the cynomolgus monkey. METHODS: Ten cyclosporine (Neoral) treated cynomolgus monkeys underwent renal allograft transplantation with bilateral nephrectomy, seven of which were examined serially with ultrasound. Ultrasound findings were compared to serum creatinine, and the results of histology from allograft biopsy on day 150 post-transplantation. RESULTS: Allografts increased in volume up to one and a half to twice that of their original volume and appeared morphologically similar to native kidneys. Allograft ureters were dilated postoperatively but decreased in size with time. Other than in two cases of ureter complications, the resistive index (RI) was normal in functioning grafts. CONCLUSIONS: Elevations in RI, as well as graft enlargement and increased cortical thickness, were related to graft pathology, but not necessarily to rejection histologically. The ultrasound findings of functioning grafts and of surgical complications after renal allograft transplantation in the cynomolgus monkey were similar to those in humans. PMID- 11396864 TI - Correlation of kidney weight and volume and selected skeletal parameters to sex in the adult rhesus monkey (Macaca mulatta). AB - This report outlines a comparison of renal weight and volume and selected skeletal parameters to sex in 22 adult male and 156 adult female rhesus macaques. Means and standard deviations for kidney weight and volume, body weight, and radiographic measurements for both males and females are reported. Ninety-five percent confidence intervals and P-values for the mean differences between the sexes for these parameters were also compiled. Male monkeys were larger, but had kidneys of similar size to those of the females. Joint distributions of the radiographic measurements of the first lumbar vertebra and the skull showed that males were larger in both measurements. The distributions of these parameters were clearly separate in males and females, while joint distributions of kidney weight and volume for males and females overlapped almost completely. We found that, regardless of age, sex, weight, or skeletal size, all normal adult rhesus monkeys generally have similar-sized kidneys. PMID- 11396865 TI - The impact of dietary protein intake on serum biochemical and haematological profiles in vervet monkeys. AB - This study evaluated the influence of Westernised and traditional African diets on biochemical and haematological profiles in vervet monkeys (Cercopithecus aethiops). Twelve adult male vervet monkeys bred at the Medical Research Council, all over 4 years of age and weighing more than 5 kg each, were divided into two groups of six individuals. These monkeys were raised on a standard in-house diet post-weaning, before they were fed for 8 weeks on diets containing milk solids (17.2%) or maize + legume (17.4%), as sources of high crude protein (+/- 3.5 g/kg). High protein diets had no significant effect on serum biochemical indices such as aspartate aminotransferase (AST) and gamma glutamyl transferase (GGT) concentrations (P > 0.10). However, alanine aminotransferase (ALT) concentrations were significantly higher during week 8 (P < 0.05) for the maize + legume protein group. Alkaline phosphatase (ALP; P < 0.07), total protein (P < 0.0001), albumin (P < 0.02), and bilirubin (P < 0.003) were elevated in the milk solids group, while glucose levels were also significantly higher for the milk solids group (P < 0.05) between weeks 2 and 6. Elevated protein intake had no significant effect on haematological parameters such as red blood cells (RBC), platelet and white blood cell (WBC) counts, haemoglobin levels and monocyte and neutrophil concentrations (P > 0.10). In contrast, serum lymphocyte levels were significantly raised in the maize + legume protein group (P = 0.03), whereas values for the haematocrit (P < 0.002), mean cell volume (MCV; P < 0.03) and mean corpuscular haemoglobin concentration (MCHC; P < 0.0001) were higher in the monkeys that were fed the milk solids. This investigation showed that the type of dietary protein that is consumed may well affect certain biochemical and haematological indices in vervet monkeys. Compared to the group that were given the traditional African food regime, the animals on the Western-type milk solids diet showed significant elevations in a number of important biological indicators. However, longer-term studies should be completed in this area if we are to make firmer conclusions regarding the link between the nature of dietary proteins that are consumed and its effect on metabolism. PMID- 11396866 TI - Therapeutic continuity in Alzheimer's disease: switching patients to galantamine. Introduction. PMID- 11396867 TI - Pharmacokinetic profiles of current therapies for Alzheimer's disease: implications for switching to galantamine. AB - Although no pharmacologic treatments have been proved to alter the pathology of Alzheimer's disease, acetylcholinesterase inhibitor (AChEI) therapy offers symptomatic improvements in or delays in the progression of cognitive, behavioral, and functional deficits. Tacrine, donepezil, rivastigmine, and galantamine are the best studied agents in this class. These drugs have varying pharmacokinetic, safety, and tolerability profiles that can affect patient outcomes. Specifically, certain metabolic parameters (ie, half-life and route of metabolism/elimination) can affect a drug's tolerability and become important when a switch from one agent to another is contemplated. The mechanism of action of galantamine, the most recently approved AChEI. varies from that of the other AChEIs in that it has allosteric modulating activity at nicotinic receptors in addition to its ability to inhibit acetylcholinesterase. Benefits of this dual mode of action on the effects of the disease have been suggested. PMID- 11396868 TI - Effects of washout and dose-escalation periods on the efficacy, safety, and tolerability of galantamine in patients previously treated with donepezil: ongoing clinical trials. AB - With the increasing number of acetylcholinesterase inhibitors (AChEIs) being marketed for the treatment of Alzheimer's disease (AD), physicians will need protocols for discontinuing one AChEI and initiating another ("switching"). Three clinical trials have been designed to provide data that will assist in the determination of the optimal conditions for switching patients from donepezil (the most widely prescribed AChEI) to galantamine (the most recently approved AChEI). The main objective of these studies is to investigate the effects of different washout periods (0 to 7 days) and dose-escalation schedules (fixed, fast vs slow) on the efficacy, safety, and tolerability of galantamine in patients with AD who were previously taking donepezil. The duration of the trials ranges from 12 to 52 weeks, and the last trial is expected to end in May 2002. No conclusions can yet be drawn from these ongoing trials, but the results should be helpful in establishing guidelines for physicians to use when switching patients with AD from donepezil to galantamine. PMID- 11396869 TI - Switching previous therapies for Alzheimer's disease to galantamine. AB - Acetylcholinesterase inhibitors (AChEIs) are the most frequently prescribed drugs for the treatment of Alzheimer's disease (AD). To date, donepezil is prescribed most often, but newer AChEIs have become available. Rivastigmine entered the pharmaceutics market for AD in 2000, and galantamine was approved for use in the United States in February 2001. Some patients with AD may already be taking a cholinesterase inhibitor, but they or their caregivers may want to change therapies for various reasons, such as lack of efficacy or poor tolerability. Therefore, defined protocols for discontinuing one therapy and initiating another therapy (ie, "switching") while maintaining efficacy and minimizing cholinergic toxicity will be essential. A post-hoc analysis of a clinical trial that enrolled patients with and without previous exposure to AChEIs indicated that the efficacy and tolerability of a second and different cholinesterase treatment were similar in both subpopulations of patients. These findings suggest that discontinuation of prior AChEI treatment is not predictive of future poor response to an effective treatment. PMID- 11396870 TI - Therapeutic continuity in Alzheimer's disease: switching patients to galantamine. Panel discussion: recommendations for prescribers. PMID- 11396871 TI - Pharmacokinetic rationale for switching from donepezil to galantamine. AB - Galantamine, the most recently approved acetylcholinesterase inhibitor (AChEI) for use in the United States, has allosteric modulating activity at nicotinic receptors and inhibits acetylcholinesterase. This dual mechanism of action may make galantamine an attractive option for patients with Alzheimer's disease who have not benefited from their current therapy; thus, methods for switching patients from donepezil or rivastigmine to galantamine are needed. Protocols for switching patients from one AChEI to another must consider both the time required for washout of the first drug and the rate of dose escalation of the second drug. Both issues depend on the pharmacodynamics, pharmacokinetics, and pharmacology of the drugs under consideration. Because the common property of the drugs considered here is their acetylcholinesterase inhibitory activity, it seems reasonable to keep this activity at or below the activity achieved by the first drug at all times. In addition, the patient's condition should be monitored to avoid deterioration resulting from subtherapeutic drug concentrations during the switch. The switching protocol proposed here has been based on an analysis of mean plasma concentrations of donepezil following administration of a single dose and on the established pharmacokinetics of galantamine. PMID- 11396873 TI - Self-organization mechanism in a bone-like hydroxyapatite/collagen nanocomposite synthesized in vitro and its biological reaction in vivo. AB - When bone is lost due to injury and/or illness, the defects are generally filled with natural bone because artificial bone materials have problems of bioaffinity. However, natural bone also has supply and infection problems. If an artificial material has the same biological properties as bone, it can replace natural bone for grafting. We synthesized a hydroxyapaite (HAp) and collagen (Col) composite by a simultaneous titration coprecipitation method using Ca(OH)2, H3PO4 and porcine atelocollagen as starting materials. The composite obtained showed a self organized nanostructure similar to bone assembled by the chemical interaction between HAp and Col. The consolidated composite by a cold isostatic pressure of 200 MPa indicated a quarter of the mechanical strength of bone. It also indicated the same biological properties as grafted bone: The material was resorbed by phagocytosis of osteoclast-like cells and conducted osteoblasts to form new bone in the surrounding area. This HAp/Col composite having similar nanostructure and composition can replace autologous bone grafts. PMID- 11396874 TI - Characterization of collagen gel solutions and collagen matrices for cell culture. AB - The influence of glutaraldehyde as a crosslinking agent to increase the strength of collagen matrices for cell culture was examined in this study. Collagen solutions of 1% were treated with different concentrations (0-0.2%) of glutaraldehyde for 24 h. The viscoelasticity of the resulting collagen gel solution was measured using dynamic mechanical analysis (DMA), which demonstrated that all collagen gel solutions examined followed the same model pattern. The creep compliance model of Voigt-Kelvin satisfactorily described the change of viscoelasticity expressed by these collagen gel solutions. These crosslinked collagen gel solutions were freeze-dried to form a matrix with a thickness of about 0.2-0.3 mm. The break modulus of these collagen matrices measured by DMA revealed that the higher the degree of crosslinking. the higher the break modulus. The compatibility of fibroblasts isolated from nude mouse skin with these collagen matrices was found to be acceptable at a cell density of 3 x 10(5) cells/cm2 with no contraction, even when using a concentration of glutaraldehyde of up to 0.2%. PMID- 11396872 TI - Taxol-loaded block copolymer nanospheres composed of methoxy poly(ethylene glycol) and poly(epsilon-caprolactone) as novel anticancer drug carriers. AB - We prepared the methoxy poly(ethylene glycol) (MePEG)/poly(epsilon-caprolactone) (PCL) amphiphilic block copolymeric nanospheres containing taxol which has promising anticancer activity. MePEG/PCL block copolymeric nanospheres (MEP50) showed a narrow size distribution and an average diameter of less than 100 nm. When the initial weight ratio of taxol to polymer was 0.5:1.0, we could obtain the nanospheres having a relatively high drug-loading of more than about 20%. The size of the MePEG/PCL nanospheres also increased according to the taxol loading. However, the nanospheres did not exhibit a significant change in the size distribution and also showed a size of less than 100 nm for even that with drug loading content (DLC) of about 20%. From the 1H NMR analysis, we identified that the MePEG/PCL nanospheres prepared by dialysis procedure have core-shell structure consisting of the hydrophilic outer shell of MePEG and the hydrophobic inner core of PCL. We confirmed the low toxicity of MePEG/PCL nanospheres (MEP70) in the acute toxicity study using male ICR mice. In addition, considering the extremely lipophilic characteristics of taxol, this MePEG/PCL, nanosphere system with high taxol loading content and suspended properties in water could be useful for the delivery of taxol. PMID- 11396875 TI - Water-based sol-gel synthesis of hydroxyapatite: process development. AB - Hydroxyapatite (HA) ceramics were synthesized using a sol-gel route with triethyl phosphite and calcium nitrate as phosphorus and calcium precursors, respectively. Two solvents, water and anhydrous ethanol, were used as diluting media for HA sol preparation. The sols were stable and no gelling occurred in ambient environment for over 5 days. The sols became a white gel only after removal of the solvents at 60 degrees C. X-ray diffraction showed that apatitic structure first appeared at a temperature as low as 350 degrees C. The crystal size and the HA content in both gels increase with increasing calcination temperature. The type of initial diluting media (i.e., water vs. anhydrous ethanol) did not affect the microstructural evolution and crystallinity of the resulting HA ceramic. The ethanol-based sol dip-coated onto a Ti substrate, followed by calcination at 450 degrees C, was found to be porous with pore size ranging from 0.3 to 1 microm. This morphology is beneficial to the circulation of physiological fluid when the coating is used for biomedical applications. The satisfactory adhesion between the coating and substrate suggests its suitability for load-bearing uses. PMID- 11396876 TI - Interlaboratory reproducibility of standard accelerated aging methods for oxidation of UHMWPE. AB - During accelerating aging, experimental uncertainty may arise due to variability in the oxidation process, or due to limitations in the technique that is ultimately used to measure oxidation. The purpose of the present interlaboratory study was to quantify the repeatability and reproducibility of standard accelerated aging methods for ultra-high molecular weight polyethylene (UHMWPE). Sections (200 microm thick) were microtomed from the center of an extruded rod of GUR 4150 HP, gamma irradiated in air or nitrogen, and circulated to 12 institutions in the United States and Europe for characterization of oxidation before and after accelerated aging. Specimens were aged for 3 weeks at 80 degrees C in an air circulating oven or for 2 weeks at 70 degrees C in an oxygen bomb (maintained at 503 kPa (5 atm.) of O2) in accordance with the two standard protocols described in ASTM F 2003-00. FTIR spectra were collected from each specimen within 24 h of the start and finish of accelerated aging, and oxidation indices were calculated by normalizing the peak area of the carbonyl region by the reference peak areas at 1370 or 2022 cm(-1). The mean relative interlaboratory uncertainty of the oxidation data was 78.5% after oven aging and 129.1% after bomb aging. The oxidation index measurement technique was not found to be a significant factor in the reproducibility. Comparable relative intrainstitutional uncertainty was observed after oven aging and bomb aging. For both aging methods, institutions successfully discriminated between air irradiated and control specimens. However, the large interinstitutional variation suggests that absolute performance standards for the oxidation index of UHMWPE after accelerated aging may not be practical at the present time. PMID- 11396877 TI - Initial interaction of osteoblasts with the surface of a hydroxyapatite poly(methylmethacrylate) cement. AB - Failure of the bone/cement interface in cemented joint prostheses is a contributor to implant loosening. The introduction of a bioactive phase, such as hydroxyapatite (HA), to cement may enhance fixation by encouraging direct bone apposition rather than encapsulation of the implant by fibrous tissue. The effect of poly(methylmethacrylate) (PMMA) bone cement (incorporating 17.5% HA wt.) on bioactivity has been investigated using primary human osteoblast-like cells (HOB). A significantly higher cell proliferation and differentiation was seen on the PMMA/HA cement compared to the PMMA cement alone, with retention of phenotype up to 21 days of culture on both materials. PMID- 11396878 TI - Specific inhibition of C3 to facilitate general complement inhibition on endotoxin affinity sorbents for apheresis applications. AB - Complement activation, as a result of human blood exposure to biomaterial surfaces, continues to be a concern in medical applications. The purpose of this study was to identify sorbent(s) and surface modification(s) that allow specific removal of endotoxin, while minimizing complement activation. Maleic anhydride (MA) modification of Sepharose CL-4B, cellulose, and Toyopearl HW-65F resulted in reduced generation of C3a, a marker of complement activation, by two orders of magnitude over unmodified surfaces. Surfaces modified with both MA and polymyxin B (PMB), utilized for binding endotoxin, reduced complement activation in a similar manner. Western Blot analysis of the larger C3 cleavage product C3dg showed a similar reduction, for all MA-modified sorbents, as observed for C3a by ELISA. C3alpha43 levels (constituent of iC3b and C3c) were also reduced, although only MA-Sepharose CL-4B levels were similar to C3a. Activation of C5, measured as the SC5b-9 complex, was also reduced by two orders of magnitude after MA modification of Sepharose CL-4B; the decrease was similar to all chemical modifications tested. PMB immobilized via CNBr on MA-modified cellulose maintained its endotoxin-binding capacity, while the latter was eliminated when PMB was immobilized via CNBr to MA-modified Sepharose CL-4B and Toyopearl HW-65F. PMID- 11396879 TI - Poly(2-hydroxyethyl methacrylate) film as a drug delivery system for pilocarpine. AB - This work investigates pilocarpine trapped in a matrix diffusion-controlled drug delivery system using hydrophilic inserts of Poly(2-hydroxyethyl methacrylate) (pHEMA) to ensure an increased bioavailability of pilocarpine and prolong the length of time in which the medication remains in the eyes of the test subjects. The physical and chemical properties of pilocarpine were investigated to elucidate the mechanism of drug-polymer interaction and the effect on drug release behavior of controlled release polymeric devices. In vitro release studies indicated that pilocarpine continued to be released from the inserts for a 24 h period. The results of intraocular pressure tests performed on albino rabbits were consistent with the observed in vitro behavior. The pressure decrease was significant for a period longer than 48 h. It confirms that the inserts, as sustainable releasing devices, are promising carriers for ophthalmic drug delivery systems. PMID- 11396880 TI - Preparation of EVAL membranes with smooth and particulate morphologies for neuronal culture. AB - In this work, the in vitro interaction of cerebellar granule neurons prepared from 7-day-old Wistar rats and poly ethylene-co-vinyl alcohol (EVAL) membranes was investigated. Cells were cultured in smooth and particulate EVAL membranes for up to 7 days. Particulate membranes were prepared by using 1-octanol to precipitate EVAL solutions in DMSO. Such a membrane was microporous characterized by a packed bed of particles. Voids left between the aggregated particles formed a continuous and interconnected porous network. Crystallization of the EVAL polymer induced by 1-octanol is responsible for the formation of particulate morphology. The membrane structure and its relationship with cells were examined by scanning electron microscopy and the MTT assay. It was observed that the particulate membrane was more favorable for the neuron culture than the smooth membrane. Neurons seeded on the particulate membrane were able to regenerate with formation of an extensive neuritic network. Therefore, the particulate structure may spatially mediate cellular response that can promote neuronal cell attachment, differentiation and neuritic growth, indicating that the particulate structure should be useful as a new polymer scaffold for nerve repair. PMID- 11396881 TI - Conversion and temperature profiles during the photoinitiated polymerization of thick orthopaedic biomaterials. AB - Polymerization of a tetrafunctional monomer was investigated under a variety of photoinitiation conditions to assess the ability to form thick materials in situ for orthopaedic applications. The major biological concerns include local cell and tissue necrosis due to the polymerization exotherm and low conversions at greater depths due to light attenuation through thick samples. Experimental results indicate that depth of cure and temperature rises are controllable by altering the photoinitiator concentration, initiating light intensity, and type of photoinitiator. For example, no measurable conversion was detected at a 1.0 cm depth when polymerization was initiated with 1.0 wt% DMPA and 100 mW/cm2 ultraviolet light, whereas approximately 40% conversion was obtained when the initiator concentration was lowered to 0.1 wt%. This conversion was further increased to approximately 55% when a photobleaching initiator system was employed. At the highest rate of initiation studied (i.e., 1.0 wt% DMPA irradiated with 100 mW/cm2 ultraviolet light), a maximum temperature of approximately 49 degrees C was reached at the sample surface; however, this temperature dramatically decreased to approximately 33 degrees C when the light intensity was decreased to 25 mW/cm2. Finally, dual initiating systems that synergistically combine the advantages of photoinitiation and thermal initiation were investigated. PMID- 11396882 TI - Visualization and detectability of elements rarely contained in soft tissue by X ray scanning analytical microscopy and electron-probe micro analysis. AB - The detectability of elements rarely contained in soft tissue was compared using X-ray scanning analytical microscope (XSAM) and electron-probe micro analysis (EPMA). Mapping images of Ca, S and P in normal soft tissue of the rat and dissolved Ni in Ni implanted soft tissue could be obtained by XSAM and EPMA. EPMA was more sensitive in detection of P, while XSAM was superior for Ca, S and Ni mapping. The high detectability for heavier elements by XSAM was explained by the large volume of characteristic X-ray generation in XSAM and low attenuation of the characteristic X-rays from heavier elements. XSAM could provide clearer mapping images for heavier elements whose concentration was low without radiation damage to specimens. PMID- 11396883 TI - Evaluation of the capacity of the SCGE assay to assess the genotoxicity of biomaterials. AB - The comet test or SCGE assay, which is already widely used in other areas, has never been used to evaluate the mutagenic potential of medical biomaterials in the final form. The purpose of our study was thus to assess the comet test as a means of assessing the genotoxic potential of finished medical biomaterials. We used silicone elastomers with increasing concentrations of 4-nitroquinoline oxide, a genotoxic agent. Hydrogen peroxide was used as the positive control, and tissue culture polystyrene as the negative control. In our study, the comet test did not detect a significant difference in genotoxicity between the pure elastomer and the same elastomer containing 0.01 mg/ml 4-nitroquinoline oxide, but did detect a significant difference between two elastomers containing 0.01 and 0.3 mg/ml of 4-nitroquinoline oxide, respectively. Since, the surface properties of the samples were identical, only the chemical composition may have caused significant differences in mutagenicity. Whatever the cause of the genotoxicity detected by the SCGE assay, testing finished biomaterials using the comet assay makes it possible to evaluate interactions between biomaterials and living tissues that are much closer to actual application conditions. PMID- 11396884 TI - In vitro preparation and ellipsometric characterization of thin blood plasma clot films on silicon. AB - The wound-healing process around implants differs from that of a normal healing without the inserted material. In this work, the composition of a natural wound surface was mimicked through clotting of a thin human blood plasma film with approximate ellipsometric thickness of 100 nm onto differently pretreated silicon surfaces. Their stability was investigated by incubations in sodium dodecyl sulphate (SDS) solutions. The enzymatic clot degradation was induced through addition of human tissue plasminogen activator (t-PA) to the plasma and the surface protein remnants after the degradation were analyzed with polyclonal antibodies. The results show that the plasma films were not SDS resistant on hydrophilic silicon. However, stability was obtained after preparation on hydrophobic silicon or when albumin or fibrinogen was immobilized to silicon before the plasma incubations. Different surfaces bound different polyclonal antibodies after the clot film degradation. The methods indicate a simple means to improve or reestablish a normal tissue inflammatory response around biomaterials. PMID- 11396885 TI - Cellular reactions and bone apposition to titanium surfaces with different surface roughness and oxide thickness cleaned by oxidation. AB - Titanium surfaces with three different surface characteristics were exposed to an intraperitoneal milieu in mouse or rat, or inserted into rabbit bone. The cleaning regimen of the TiO2 surfaces in this study included oxidation by heat or acid and a final rinsing and storage in water. Intraperitoneal exposure ranged from 1 to 64 min and the healing period in bone was 6 weeks. Cell recruitment to the surfaces was quantified by acridine orange staining and specific antibodies directed against cell membrane antigens. Removal torque, bone-to-metal contact, total bone area and histological evaluations were used to evaluate fixture stability and the healing-in of the implants. After the healing period of 6 weeks only a transient significant difference was seen in the total number of cells adherent on the surfaces. No significant differences were observed between any of the surfaces for removal torque, bone-to-metal contact, or bone area. The areas lacking bone-to-metal contact were filled with normal vascularised connective tissue with no signs of fibrous capsule formation or giant cells. These findings differ from findings published earlier of Ti implants that underwent a cleaning regimen with alcohol as the final rinsing step. The tissues around the implants were richly vascularised and there was continued bone growth toward the surfaces. The bone-to-metal contact in this study was lower than that seen with alcohol cleaned TiO2. PMID- 11396886 TI - Influence of mixing techniques on the physical properties of acrylic bone cement. AB - Palacos R bone cement was prepared using three commercially available mixing techniques, first generation, second generation and third generation, to determine the mechanical properties and porosity contents of the bone cement. The compressive strengths, bending strengths and flexural moduli were expressed as a function of void content. The volume of pores within the cement structure was found to be a contributing factor to the physical properties of acrylic bone cement. The lower the volume of voids in the cement the better the compressive and flexural properties, hence stronger bone cement. It was found that the best results were obtained from cement that had been mixed using the Mitab Optivac or Summit HiVac Syringe systems at a reduced pressure level of between -72 and -86 kPa below atmospheric pressure, resulting in cement of porosity 1.44-3.17%; compressive strength 74-81 MPa; flexural modulus 2.54-2.60 GPa; and flexural strength 65-73 MPa. PMID- 11396887 TI - Influence of residual stress on bonding strength and fracture of plasma-sprayed hydroxyapatite coatings on Ti-6Al-4V substrate. AB - Six hydroxyapatite coatings (HACs) were plasma sprayed on Ti-6Al-4V substrates by varying the substrate temperatures and the cooling conditions. This study is aimed not only to measure the residual stress of HACs under various conditions, but also to assess the influence of residual stress in HACs on their bonding strength. The residual stress and bonding strength were measured by XRD "sin2 psi" technique and standard adhesion test (ASTM C-633), respectively. The result of the study clearly established the relationship between bonding strength and residual stress. The arguments leading to the above conclusion were discussed in detail. Fractographic analysis indicated that fracture of the system occurred mainly inside the hydroxyapatite coating under lower residual stress; as residual stress increased, fracture tended to occur more easily along the crucial HA-Ti alloy substrate interface. A mechanism was presented for the relationships between residual stress, fracture behaviour and bonding strength for the plasma sprayed hydroxyapatite coatings on Ti-6Al-4V substrate. PMID- 11396888 TI - Mitigation of bioprosthetic heart valve degeneration through biocompatibility: in vitro versus spontaneous endothelialization. AB - BACKGROUND: Glutaraldehyde-related cytotoxicity and transanastomotic ingrowth inhibition prevent the spontaneous endothelialization of bioprosthetic heart valves. In order to evaluate the ability of improved biocompatibility to reduce tissue degeneration, conventionally fixed aortic root prostheses were both glutaraldehyde-detoxified and in vitro endothelialized. METHODS: Entire aortic roots were fixed in 0.2% glutaraldehyde (GA) (control group) and either detoxified in acetic acid-buffered urazole (0.1 M) or detoxified and in vitro lined with cultured, autologousjugular vein endothelial cells. The valved roots were inserted in the distal aortic arch of 15 juvenile Merino sheep for a period of 12 weeks. Upon explant, leaflets, sinuses and aortic wall of the prostheses were analysed by SEM to assess the surface endothelium, histologically regarding tissue inflammation, and by atomic absorption spectrophotometry to determine the content of tissue calcium. RESULTS: There was no endothelium on control grafts, except for a short anastomotic pannus. The detoxified group showed an incomplete patchy endothelium on the aortic wall but hardly any on the leaflets, whereas, the in vitro lined group had aortic wall, sinuses and most of the leaflets confluently endothelialized. Tissue inflammation was prominent in the control group and least expressed in the endothelialized group (p < 0.05). Detoxification significantly reduced leaflet calcification. In the aortic wall, both detoxification and endothelial lining were required to significantly mitigate calcification. CONCLUSION: In the 12 week circulatory sheep model, the calcium mitigating effect of detoxification was more pronounced than that of in vitro endothelialization. Nevertheless, there was a distinct overall benefit if detoxification was combined with endothelialization. PMID- 11396889 TI - Colloidal processing of hydroxyapatite. AB - Reliable bioceramics are needed to implement the high requirements that living tissues demand. This work focuses on the processing steps necessary to manufacture advanced ceramics that can be used as implant devices. The influence of the heat treatment temperature on the characteristics of a precipitated hydroxyapatite (OHAp) powder was evaluated in order to obtain an appropriate specific surface area for colloidal processing. Ball milling of the calcined powders for 20 h was required to improve the rheological properties of the suspensions and the packing ability during consolidation. Different dispersing agents were tested and the first trial was made based on their effect on the zeta potential. The most promising ones were then selected and their efficiency was evaluated from rheological measurements and slip-casting performance of suspensions prepared at different solids loading. Targon 1128 was revealed to be the most efficient dispersant, enabling to prepare fluid suspensions containing 50 vol% solids and the highest green and sintered density values to be obtained. PMID- 11396891 TI - The gastrointestinal transit profile of 14C-labelled poly(acrylic acids): an in vivo study. AB - The gastrointestinal distribution profiles for three 14C-labelled poly(acrylic acid)s of different average molecular weights and degrees of cross-linking have been established using the rat model. Despite initial differences in transit times and retention characteristics, these structural features were found to be of little influence to the overall gastrointestinal transit of the materials under consideration. No evidence for the systemic absorption of any of the polymers could be identified. PMID- 11396890 TI - Corrosion behavior of titanium nitride coated Ni-Ti shape memory surgical alloy. AB - Nickel-titanium (NiTi, nitinol) shape memory alloy was nitrided using an original powder immersion reaction assisted coating (PIRAC) method in order to modify its surface properties. PIRAC nitriding method is based on annealing the samples in the atmosphere of highly reactive nitrogen supplied by decomposition of unstable nitride powders or, alternatively, by selective diffusion of the atmospheric nitrogen to the sample surface. Being a non-line-of-sight process, PIRAC nitriding allows uniform treatment of complex shape surgical implants. Hard two layer titanium nitride (TiN)/Ti2, Ni coatings were obtained on NiTi surface after PIRAC anneals at 900 and 1000 degrees C. PIRAC coating procedure was found to considerably improve the corrosion behavior of NiTi alloy in Ringer's solution. In contrast to untreated nitinol, no pitting was observed in the samples PIRAC nitrided at 1000 degrees C, 1 h up to 1.1 V. The coated samples were also characterized by very low anodic currents in the passive region and by an exceedingly low metal ion release rate. The research results suggest that PIRAC nitriding procedure could improve the in vivo performance of NiTi alloys implanted into the human body. PMID- 11396892 TI - Self-expandable chitosan stent: design and preparation. AB - Stents are largely used in surgical procedures to relieve pathological obstructions. The purpose of the present study was to design and prepare a biocompatible stent with a self-expandable mechanism. Thin films were prepared from deacetylated chitosan (4% w/v) dissolved in acetic acid solution (2% v/v). The chitosan films were tested by a calibrated tensiometer to measure the Young's module (E). The films were used to manufacture stents by pulling and winding them around a cylindrical rod in a helical fashion. Thirteen stents (diameter = 0.5 +/ 0.05 mm, length approximately 4 mm) were inserted into the vas deferens of wistar rats. Upon stent insertion, the vasal anastomosis was achieved with a laser-soldering technique. The animals were sacrificied 8 weeks later. The stress test showed that the chitosan film was elastic (maximum strain = 105% +/- 6%, E = 0.7655 +/- 0.0288 Mpa). The stents self-expanded by releasing their elastic energy. All the stents but one remained open inside the vasa despite high incidence of sperm granuloma. A biocompatible and self-expandable stent with a helical design is proposed. PMID- 11396893 TI - A small punch test technique for characterizing the elastic modulus and fracture behavior of PMMA bone cement used in total joint replacement. AB - Polymethylmethacrylate (PMMA) bone cement is used in total joint replacements to anchor implants to the underlying bone. Establishing and maintaining the integrity of bone cement is thus of critical importance to the long-term outcome of joint replacement surgery. The goal of the present study was to evaluate the suitability of a novel testing technique, the small punch or miniaturized disk bend test, to characterize the elastic modulus and fracture behavior of PMMA. We investigated the hypothesis that the crack initiation behavior of PMMA during the small punch test was sensitive to the test temperature. Miniature disk-shaped specimens, 0.5 mm thick and 6.4 mm in diameter, were prepared from PMMA and Simplex-P bone cement according to manufacturers' instructions. Testing was conducted at ambient and body temperatures, and the effect of test temperature on the elastic modulus and fracture behavior was statistically evaluated using analysis of variance. For both PMMA materials, the test temperature had a significant effect on elastic modulus and crack initiation behavior. At body temperature, the specimens exhibited "ductile" crack initiation, whereas at room temperature "brittle" crack initiation was observed. The small punch test was found to be a sensitive and repeatable test method for evaluating the mechanical behavior of PMMA. In light of the results of this study, future small punch testing should be conducted at body temperature. PMID- 11396894 TI - Preparation of nanoparticles composed with bioinspired 2-methacryloyloxyethyl phosphorylcholine polymer. AB - The poly(L-lactic acid) nanoparticles immobilized with 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer, which has excellent blood compatibility, were prepared by a solvent evaporation technique using the water-soluble amphiphilic MPC polymer as an emulsifier and a surface modifier. The diameter and zeta potential of the obtained nanoparticles strongly depended on the concentration of the MPC polymer. When the nanoparticles were prepared in 1.0 mg/ml of an MPC polymer aqueous solution, the diameter was 221 nm which was determined by atomic force microscopy and dynamic light scattering measurements. The X-ray photoelectron spectroscopic analysis indicated that the phosphorylcholine groups of the MPC unit were located at the surface of the nanoparticles, that is, the MPC polymer was immobilized on the PLA particles and the surface zeta-potential was -2.5 mV. Various hydrophobic fluorescence probes could permeate through the MPC polymer layer and adsorb on the PLA surface. The amount of bovine serum albumin adsorbed on the nanoparticles was significantly smaller compared with that on the conventional polystyrene nanoparticles. It is suggested that the nanoparticles immobilized with the MPC polymer have the potential for use as both a novel drug carrier and diagnostic reagent which can come in contact with blood components. PMID- 11396895 TI - Formation of dextran hydrogels by crystallization. AB - In this paper, a novel method is presented for the preparation of dextran hydrogels and microspheres, based on crystallization. Although dextrans are known to be well soluble in water, precipitation was observed in concentrated aqueous solutions of low molecular weight dextran (dextran 6000), whereas for solutions of dextran with higher molecular weights (dextran 40,000 and 220,000) no precipitation was observed in the time-frame studied. The kinetics of the precipitation process were studied and showed that precipitation was faster when more concentrated dextran solutions were used. Furthermore, the precipitation process was accelerated by stirring and by the presence of salts. Depending on the precipitation time, microspheres or gels were obtained. The precipitates were insoluble in water at room temperature, but readily dissolved in boiling water or DMSO. IR spectroscopy and (modulated) differential scanning calorimetry ((M)DSC) demonstrated that the precipitates were crystalline. We hypothesize that crystallization is due to association of the chains through hydrogen bonding, induced by the large polymer/water ratio in concentrated dextran 6000 solutions. PMID- 11396896 TI - A crosslinked system from scleroglucan derivative: preparation and characterization. AB - Matrices obtained by a crosslinking reaction between the polycarboxylated derivative of scleroglucan (sclerox) and 1,6-hexanedibromide have been prepared and characterized. Different ratios between the alkane dihalide and sclerox yielded products with different properties. Water uptake by the hydrogel with a low degree of crosslinking was remarkably affected by ionic strength. The determination of the crosslink density is led by simultaneously solving two Flory equilibrium equations referring to two different conditions characterized by the presence or the absence of a salt in the swelling agent. Moreover, the swelling kinetics was studied by means of a recently proposed model. Finally, the permeation of two model molecules (theophylline and polystyrene sulphonate-sodium salt) through the hydrogels was evaluated. PMID- 11396897 TI - Cytocompatibility and response of osteoblastic-like cells to starch-based polymers: effect of several additives and processing conditions. AB - This work reports on the biocompatibility evaluation of new biodegradable starch based polymers that are under consideration for use in orthopaedic temporary applications and as tissue engineering scaffolds. It has been shown in previous works that by using these polymers it is both possible to produce polymer/hydroxyapatite (HA) composites (with or without the use of coupling agents) with mechanical properties matching those of the human bone, and to obtain 3D structures generated by solid blowing agents, that are suitable for tissue engineering applications. This study was focused on establishing the influence of several additives (ceramic fillers, blowing agents and coupling agents) and processing methods/conditions on the biocompatibility of the materials described above. The cytotoxicity of the materials was evaluated using cell culture methods, according to ISO/EN 109935 guidelines. A cell suspension of human osteosarcoma cells (HOS) was also seeded on a blend of corn starch with ethylene vinyl alcohol (SEVA-C) and on SEVA-C/HA composites, in order to have a preliminary indication on cell adhesion and proliferation on the materials surface. In general, the obtained results show that all the different materials based on SEVA-C, (which are being investigated for use in several biomedical applications), as well as all the additives (including the novel coupling agents) and different processing methods required to obtain the different properties/products, can be used without inducing a cytotoxic behaviour to the developed biomaterials. PMID- 11396898 TI - Lysine-derivatized polyurethane as a clot lysing surface: conversion of adsorbed plasminogen to plasmin and clot lysis in vitro. AB - Polyurethane surfaces to which lysine residues are immobilized by photochemical methods are proposed as a basis for clot lysing surfaces. The lysines are attached in such a way that the epsilon-amino and carboxyl groups are free. We showed previously that these surfaces, when placed in contact with plasma, adsorb only plasminogen and virtually no other proteins (McClung et al., J. Biomed. Mater. Res. 49 (2000) 409). In this communication, data based on a chromogenic substrate assay are presented showing that plasminogen adsorbed to these surfaces is readily converted to plasmin in the presence of tissue-plasminogen activator (t-PA). Moreover, the rate of activation on the surface is considerably greater than in solution. Experiments demonstrating the ability of these surfaces to dissolve fibrin clots are also reported. Surfaces exposed to plasma and then to t PA were placed in citrated plasma. On recalcification, clotting was initiated, but the incipient clots were soon dissolved. On control surfaces (no lysine or lysine in which the epsilon-amino groups were not available) coagulation continued until a stable clot was formed. Similar observations were made when the plasma/t-PA exposed surfaces were placed in a pure fibrinogen solution and thrombin was added. PMID- 11396899 TI - Spot compliant neuronal networks by structure optimized micro-contact printing. AB - Neuronal cell growth in vitro can be controlled with micropatterned structures of extracellular matrix proteins such as laminin. This technique is a powerful tool for studying neuronal cell function in order to increase experimental reproducibility and to specifically design innovative experimental setups. In this paper the correlation between the structural dimensions of the ECM pattern and the shape of the resulting cellular network is analyzed. The aim of the present study was to position neuronal cell bodies as precisely as possible and to induce directed cell differentiation. PCC7-MzN cells were cultured on laminin patterns. The line width, node size and gap size in-between cell adhesion sites was varied systematically. Micrographs of the samples were taken and statistically analyzed using Student's t-test and linear correlation methods. Precise cell positioning has successfully been performed and evidence for controlled neuronal polarization has been found. With a structure geometry of 4 microm line width, 20 microm node size and 10 microm gap size a nodal compliance of 86% (+/- 10%) has been achieved. PMID- 11396900 TI - A self-setting single-component calcium phosphate cement. AB - Following an original synthesis route, we have prepared a single-component calcium phosphate apatite-like powder which settles and hardens when mixed with deionized water in an approximative 1.2 g:1 ml ratio. This paper describes the first physico-chemical studies and characterizations of the material. Observations of its in vitro behavior show a slight volume contraction and toxicity against fibroblasts bone marrow cells on disks of compacted powder. It is suggested that after an improvement of the powder characters such as grain size, and the choice of another hardening liquid, to name a few, this material should be a potential--or an ingredient of--bone cement. PMID- 11396901 TI - State observers for a biological wastewater nitrogen removal process in a sequential batch reactor. AB - Biological removal of nitrogen is a two-step process: aerobic autotrophic microorganisms oxidize ammoniacal nitrogen to nitrate, and the nitrate is further reduced to elementary nitrogen by heterotrophic microorganisms under anoxic condition with concomitant organic carbon removal. Several state variables are involved which render process monitoring a demanding task, as in most biotechnological processes, measurement of primary variables such as microorganism, carbon and nitrogen concentrations is either difficult or expensive. An alternative is to use a process model of reduced order for on-line inference of state variables based on secondary process measurements, e.g. pH and redox potential. In this work, two modeling approaches were investigated: a generic reduced order model based on the generally accepted IAWQ No. 1 Model [M. Henze, C.P.L., Grady, W., Gujer, G.V.R., Marais, T., Matsuo, Water Res. 21 (5) (1987) 505-515]-generic model (GM), and a reduced order model specially validated with the data acquired from a benchscale sequential batch reactor (SBR) specific model (SM). Model inaccuracies and measurement errors were compensated for with a Kalman filter structure to develop two state observers: one built with GM, the generic observer (GO), and another based on SM, the specific observer (SO). State variables estimated by GM, SM, GO and SO were compared to experimental data from the SBR unit. GM gave the worst performance while SM predictions presented some model to data mismatch. GO and SO, on the other hand, were both in very good agreement with experimental data showing that filters add robustness against model errors, which reduces the modeling effort while assuring adequate inference of process variables. PMID- 11396902 TI - Characterization of de-emulsification capabilities of a Micrococcus species. AB - Effect of post harvest washing as well as cell concentration on de-emulsification characteristics of an isolated Micrococcus species has been tested with Tween 60 Span 60 stabilised oil in water (o/w) and L-92 pluronic surfactant stabilised water in oil (w/o) model emulsions (kerosene water). The cells used were 140 h old and grown under submerged conditions at 37 degrees C in a medium containing n tetradecane (4% v/v) as the carbon source. The harvested bacterial cells when in an unwashed condition (at a cell concentration of 2 mg/ml of emulsion) were found to de-emulsify the o/w system at a much faster rate than the w/o system exhibiting half-life values for the respective system as 10.2 and 127.7 h. Post harvest washing of the cells with any lipid solubilising solvent (n-pentane, n hexane, kerosene, chloroform-methanol-water (CMW)) yielded a decrease in their de emulsification power for w/o emulsion. But the decay of o/w emulsion became faster with n-pentane- and kerosene-washed cells as evident from their corresponding half-life values of 3.3 and 4.6 h. Compared to the w/o system, an increase in the concentration of kerosene-washed cell had a direct effect on de emulsification for the o/w system. For cell concentrations of 2, 3 and 4 mg/ml of the emulsion, the half-life values for the w/o system were 364.8, 442.0 and 454.9 h, respectively. For 2 and 4 mg/ml cell contents, the half-life values for the o/w system were 4.6 and 1.0 h. The decay of both the emulsions was very slow or even incomplete for cell concentrations less than 2 mg/ml. De-emulsifying capacity of the n-tetradecane grown Micrococcus species towards o/w model emulsion improved considerably after washing the cells with n-pentane and kerosene, and use of kerosene-washed cells (4 mg/ml) reduced the half-life to 1 h. PMID- 11396903 TI - Influence of the operating conditions in the acetone pulping of wheat straw on the properties of the resulting paper sheets. AB - A central composite factor design was used to examine the influence of independent variables in the acetone pulping of wheat straw (processing temperature, time, and acetone concentration) on the yield of the resulting pulp, and on various physical properties of paper sheets (breaking length, stretch, burst index, tear index and brightness) obtained from it. Equations that related each dependent variable to the different independent variables were obtained, and these reproduced the experimental results for the yield, breaking, stretch, burst index and brightness obtained at temperatures, times, and acetone concentratons over the ranges 140-180 degrees C, 60-120 min and 40-80%, respectively, with errors less than 20%. Obtaining the optimum breakinig length, stretch, burst index and tear index for the paper sheets (3,456 m, 1.42%, 1.36 kN/g and 3.86 mNm2/g, respectively) entails using a high temperature; the processing time and acetone concentration only influence stretch, optimization of which requires using a short time and a low concentration. The optimum brightness (30.44%) is achieved with a low temperature, a short time and a medium acetone concentration. In order to minimize losses of solvent during its recovery and recycling while ensuring acceptable levels of the properties of the paper sheets, a high temperature, a low acetone concentration and a short time can be used; the brightness level thus obtained is only 10% lower than the optimum value. PMID- 11396904 TI - Microbial decomposition of post-harvest sugarcane residue. AB - A laboratory in situ composting study was conducted as a possible alternative method for the current practice of open air burning of post-harvest sugarcane residue by sugarcane farmers. In situ composting of the sugarcane residue by the indigenous bacteria and fungi was accelerated using molasses as an initial substrate. A one-time application of molasses boosted the soil microbial population. which started to decompose the ligno-cellulosic fractions of the residue. The study showed significant differences in several parameters among the control and molasses applied treatments, namely, visual decomposition of residue, bacterial and fungal population, soil pH, cellulose content, cellulase activity. and soil organic matter. Further study is needed to refine the process for the future application of this technology as a possible alternative to the current practice of open air burning of sugarcane residue by farmers. PMID- 11396905 TI - Potential utilisation of sewage sludge and paper mill waste for biosorption of metals from polluted waterways. AB - The adsorption of cadmium, copper(II), lead and zinc ions from aqueous solution by sewage sludge, paper mill waste (PMW) and composted PMW was investigated along with the influence of pre-treatment on composted PMW. Langmuir adsorption isotherms were fitted where appropriate. Sewage sludge was the most effective biosorbent of the waste products for all metal ions examined, adsorbing, for example, up to 39.3 mg/g of Pb at an initial concentration of 77.8 mg/l. PMW was a less effective biosorbent than sewage sludge. However, it was found that composting the PMW resulted in an increase in metal uptake capacity and both sewage sludge and composted PMW have potential for low-cost remediation of high leachate wastewaters. The desorption of metal ions from PMW compost was most effective using 0.1 N H2SO4 and 1 mM nitrilotriacetic acid (NTA). PMID- 11396906 TI - Effects of rhizobacteria and root symbionts on the reproduction of Meloidogyne javanica and growth of chickpea. AB - The effects of rhizobacteria, i.e. Pseudomonas fluorescens, Azotobacter chyroococcum and Azospirillum brasilense, alone and in combination with root symbionts, Rhizobium sp. and Glomus mosseae, on the growth of chickpea, Cicer arietinum, and reproduction of Meloidogyne jaranica were studied. When added alone G. mosseae was better at improving plant growth and reducing galling and nematode reproduction than any other tested organism. Application of P. fluorescens caused an almost similar increase in plant growth to that caused by Rhizobium sp., while use of A. chroococcum was better than A. brasilense in improving growth of nematode --infected plants. Combined use of P. fluorescens with G. mosseae was better at improving plant growth and reducing galling and nematode multiplication than any other combined treatment. PMID- 11396907 TI - Production and separation of alpha-agarase from Altermonas agarlyticus strain GJ1B. AB - This paper presents results on the production of alpha-agarase by a fermentation process and its separation using membrane microfiltration (MF). Optimization of fermentation conditions for alpha-agarase production using Altermonas agarlyticus grown on medium containing agar as a carbon source was done in batch, fed-batch and continuous fermentations. Continuous culture at a dilution rate of 0.03 h(-1) appeared to be best suited for production of alpha-agarase by this organism. At 0.03 h(-1) dilution rate, enzyme activity was 0.9 U/ml. Clarification of broth was done using a hollow-fibre microfiltration membrane. The influence of hydrodynamic parameters on permeate flux and enzyme activity was studied. The best performance was obtained with prefiltered fermentation broth. A stable permeate flux of about 250-270 ml/min.m2 and an enzyme retention rate between 0% and 25% was obtained at temperatures between 6 degrees C and 22 degrees C, transmembrane pressure of 100 mm Hg and fluid cross-flow velocity of 4 x 10(-2) m/s. From the experiments on concentration of fermentation broth, the best compromise between enzyme activity transmission and permeate flux was obtained at a concentration factor of 2. PMID- 11396909 TI - Preparation of levoglucosan by pyrolysis of cellulose and its citric acid fermentation. AB - Levoglucosan (LG), 1,6-anhydro-beta-D-glucopyranose, was produced by pyrolysis of cellulose. A response surface method was used to optimize the reaction parameters: X1, temperature; X2, time required for heating cellulose from room temperature to the designed pyrolysis temperature; and X3, vacuum, and a Box Behnken design was employed for this purpose. The optimal temperature and time were found to be 388 degrees C and 26.2 min by fixing the vacuum at 1 mm Hg. The levoglucosan prepared was fermented to citric acid by Aspergillus niger CBX-209, which was a mutant derived by gamma-ray mutagenesis of the parent strain CBX-2. The mutant could produce increasing citric acid with increasing LG purity and had a citric acid yield of 87.5% when using purified levoglucosan as the sole carbon source in a 5 day fermentation period. PMID- 11396908 TI - Steam-explosion of olive stones: hemicellulose solubilization and enhancement of enzymatic hydrolysis of cellulose. AB - Olive stones (whole stones and seed husks in fragments) were processed by steam explosion under different experimental conditions of temperature and time, 200 236 degrees C for 2-4 min, with or without previous acid impregnation with 0.1%, H2SO4 (w/w). This paper examines the solubilization of hemicelluloses and their molecular weight distribution. The subsequent enzymatic hydrolysis of the solid residue, using a preparation of cellulase, was also studied. The maximum yield of the pentosan recovered in the water solution was 63% pentose in the starting material for seed husk treated at 200 degrees C for 2 min (log R0 3.24) prior to acid-impregnation, or at 215 degrees C for 2 min (log R0 3.69) without acid, compared to 39% of the potential yield for whole stones pre-impregnated with acid under more severe conditions (at log R0 = 4.07). This indicates that the autohydrolysis of hemicellulose in seed husks when compared to whole stones is enhanced. The molecular weight distribution of profile sugars showed that the depolymerization of hemicelluloses is a function of the severity of the treatment. Steam-explosion improved the accessibility of the cellulose and increased the enzymatic hydrolysis yield after steam-explosion with respect to material without steam explosion (ball-milled material), although little increase in the extent of saccharification occurred when the alkali-soluble lignin was removed. Only when the substrate was post-treated with Na-chlorite was the enzymatic hydrolysis improved, the water-insoluble residue being almost completely hydrolyzed in 8 h of incubation. PMID- 11396910 TI - Towards maximising output from vermireactors fed with cowdung spiked paper waste. AB - Paper waste, spiked with varying proportions of cowdung, was vermicomposted in 'low-rate' and 'high-rate' reactors. The former type of reactors had earthworm populations and feed loading rates similar to ones recommended by previous workers. The 'high-rate' reactors were operated with 12.5 times higher earthworm densities and feed loading rates. All the reactors were studied for six months to assess the vermicast output, survivability, growth and reproduction of the earthworms hence the sustainability of the reactors--for long-term, continuous operation. The studies revealed the viability of the high-rate vermireactor concept. The high-rate reactors consistently produced over 6.5 times more castings per unit digester volume with no adverse effect on the earthworm population, as reflected by (a) absence of mortality, (b) consistent growth in worm zoomass, and (c) normal rate of reproduction. The studies also revealed that an increase in the cowdung fraction in the feed from 14.3% to 20% (4:1 paper:cowdung blends to 6:1 blends) had little positive impact on the vermicast output or earthworm health. This indicated that spiking of paper feed with approximately 14% cowdung, or perhaps an even smaller fraction, might be adequate to support earthworms in the paper-fed vermireactors. PMID- 11396911 TI - Biosorption of Cr (VI) from aqueous solution by Rhizopus nigricans. AB - The study was aimed to quantify the Cr sorption ability of powdered biomass of Rhizopus nigricans at the best operating conditions. The influence of solution pH, agitation, Cr (VI) concentration, biomass dosage, contact time, biomass particle size and temperature were studied. The optimum pH for biosorption of Cr (VI) was found to be 2.0. Higher adsorption percentage was noted at lower initial concentrations of Cr ions, while the adsorption capacity of the biomass increased with increasing concentration of ions. Optimum biomass dosage was observed as 0.5% (w/v). More than 75% of the ions were removed within 30 min of contact and maximum removal was obtained after 8 h. Biomass particles of smaller size (90 microm) gave maximum adsorption (99.2%) at 100 mg/l concentration. The adsorption capacity increased with increase in temperature and agitation speed and the optimum were determined as 45 degrees C at 120 rpm. Freundlich and Langmuir isotherms were used to evaluate the data and the regression constants were derived. The adsorption rate constant values (Kad) were calculated for different initial concentration of Cr ions and the sorption was found to be higher at lower concentration (100 mg/l) of metal ion. PMID- 11396912 TI - Improvement of biogas production from vinasse via covalently immobilized methanogens. AB - Improvement of biogas production was realized by covalent immobilization of a methanogenic consortium onto a granulated polymeric support [poly(acrylonitrile acrylamide)]. The growth kinetics of the immobilized consortium was investigated during a process of vinasse methanation, and a cell concentration increase from 12.3 mg g(-1) support to 52.1 mg g(-1) support was established. The methane yield reached 0.33 m3 kg(-1) CODr, the maximum yield on chemical oxygen demand (COD) removal being 92%. The inhibitory effect of oxygen was reduced by immobilizing the methanogenic consortium. PMID- 11396913 TI - A preliminary investigation into the biofuel characteristics of tigernut (Cyperus esculentus) oil. AB - Laboratory scale experiments were carried out to produce and characterise biofuel from tigernut (Cyperus esculentus) oil. Transesterification of tigernut oil afforded methyl and ethyl esters, which had fuel properties similar to common biofuels, hence tigernut could be utilised as an alternative renewable energy resource. PMID- 11396914 TI - Improvement of the physical properties of reprocessed paper by using biological treatment with modified cellulase. AB - A primary need for waste paper reprocessing is to preserve optical properties and the physical strength of the paper fibers. In this study, modified cellulase with copolymer, polyethylene oxide (PEO) derivatives and maleic anhydride (MA) was applied to the reprocessing of mixed office waste (MOW). Modified cellulase was prepared by a chemical reaction between amino groups of the cellulase and the MA functional groups of the copolymer. In MOW reprocessing, modified cellulase improved several physical properties of the paper including freeness, optical properties, and physical strength compared to the conventional process. Even though native cellulase improved the physical properties, paper treated with modified cellulase exhibited an increase in physical properties such as tensile strength and internal bond over those of unmodified cellulase. From these results, modified cellulase method is a new biological treatment that will save pulp resources, which are added to waste paper reprocessing to maintain the strength of paper. PMID- 11396915 TI - Evaluation of various agro-wastes for traditional black soap production. AB - The agricultural wastes, cocoa-pod husks, palm-bunch waste, sorghum chaff and groundnut shells, which are normally thrown away have been used in the production of black soap. Unlike other soaps which are made from oils and chemicals, black soap is made from oils and agro-wastes ashes. Chemical analysis indicated that the liquid extract from the ashes of the different agro-wastes used contained various amounts of potassium and sodium compounds. The most common ingredient in the agro-wastes was potassium carbonate. The amount of potassium carbonate was 56.73 +/- 0.16% in cocoa-pod ash, 43.15 +/- 0.13% in palm-bunch ash, 16.65 +/- 0.05% in groundnut shell ash and 12.40 +/- 0.08% in sorghum chaff ash. Soaps made from the agro-wastes ashes had excellent solubility, consistency, cleansing and lathering abilities. PMID- 11396916 TI - Inhibition effect of initial Pb2+ concentration on Pb2+ accumulation by Saccharomyces cerevisiae and Aureobasidium pullulans. AB - Pb2+ accumulation by Saccharomyces cerevisiae and Aureobasidium pullulans was inhibited by the initial Pb2+ concentration. In the case of S. cerevisiae, as initial Pb2+ concentrations increased, the accumulated Pb2+ per unit cell dry weight at equilibrium and the time required to reach an equilibrium state increased at low initial Pb2+ concentration. On the contrary, the accumulated Pb2+ decreased at high initial Pb2+ concentration at all pH values. The inhibition effect of initial Pb2+ concentration was delayed by the decrease of pH. However, the maximal Pb2+ accumulation capacity of S. cerevisiae was almost constant regardless of pH values. In the case of A. pullulans, the time required to reach an equilibrium state was independent of the initial Pb2+ concentration. The maximal Pb2+ accumulation capacity of A. pullulans decreased according to the decrease of pH values. However, the initial Pb2+ concentration needed to reach maximal Pb2+ accumulation amount was almost constant. PMID- 11396917 TI - Structure and function of sulfotransferases. AB - Sulfotransferases (STs) catalyze the transfer reaction of the sulfate group from the ubiquitous donor 3'-phosphoadenosine 5'-phosphosulfate (PAPS) to an acceptor group of numerous substrates. This reaction, often referred to as sulfuryl transfer, sulfation, or sulfonation, is widely observed from bacteria to humans and plays a key role in various biological processes such as cell communication, growth and development, and defense. The cytosolic STs sulfate small molecules such as steroids, bioamines, and therapeutic drugs, while the Golgi-membrane counterparts sulfate large molecules including glucosaminylglycans and proteins. We have now solved the X-ray crystal structures of four cytosolic and one membrane ST. All five STs are globular proteins composed of a single alpha/beta domain with the characteristic five-stranded beta-sheet. The beta-sheet constitutes the core of the Paps-binding and catalytic sites. Structural analysis of the PAPS-, PAP-, substrate-, and/or orthovanadate (VO(3-)(4))-bound enzymes has also revealed the common molecular mechanism of the transfer reaction catalyzed by sulfotransferses. The X-ray crystal structures have opened a new era for the study of sulfotransferases. PMID- 11396918 TI - Inhibitors to the Src SH2 domain: a lesson in structure--thermodynamic correlation in drug design. AB - Src homology 2 (SH2) domains play a key role in many tyrosine kinase-mediated intracellular signal transduction pathways. Aberrancies in the interaction of these domains can lead to a range of disease states. As a result, the pharmaceutical industry has made a large temporal and financial investment in the development of specific inhibitors to these domains. Focusing on the interactions of the SH2 domain from the protein Src, we report how the correlation of structural and thermodynamic data allows an assessment of the process of drug design. The binding site of the protein includes two pockets; one interacts with phosphotyrosine groups on cognate ligands, and the other accommodates an aliphatic hydrophobic side chain. The interaction with cognate ligands is also mediated by a network of water molecules. Thermodynamic data from isothermal titration calorimetric studies suggest that modification of the interactions in the SH2 binding site has been largely unsuccessful in producing high-affinity inhibitors. Furthermore, it appears that compounds that disrupt the interfacial water pay the price for the loss of the contribution to the free energy from a network of hydrogen bonds. PMID- 11396919 TI - The binding energetics of first- and second-generation HIV-1 protease inhibitors: implications for drug design. AB - KNI-764 is a powerful HIV-1 protease inhibitor with a reported low susceptibility to the effects of protease mutations commonly associated with drug resistance. In this paper the binding thermodynamics of KNI-764 to the wild-type and drug resistant mutant V82F/I84V are presented and the results compared to those obtained with existing clinical inhibitors. KNI-764 binds to the wild-type HIV-1 protease with very high affinity (3.1 x 10(10) M(-1) or 32 pM) in a process strongly favored by both enthalpic and entropic contributions to the Gibbs energy of binding (Delta G = -RTlnK(a)). When compared to existing clinical inhibitors, the binding affinity of KNI-764 is about 100 fold higher than that of indinavir, saquinavir, and nelfinavir, but comparable to that of ritonavir. Unlike the existing clinical inhibitors, which bind to the protease with unfavorable or only slightly favorable enthalpy changes, the binding of KNI-764 is strongly exothermic (-7.6 kcal/mol). The resistant mutation V82F/I84V lowers the binding affinity of KNI-764 26-fold, which can be accounted almost entirely by a less favorable binding enthalpy to the mutant. Since KNI-764 binds to the wild type with extremely high affinity, even after a 26-fold decrease, it still binds to the resistant mutant with an affinity comparable to that of other inhibitors against the wild type. These results indicate that the effectiveness of this inhibitor against the resistant mutant is related to two factors: extremely high affinity against the wild type achieved by combining favorable enthalpic and entropic interactions, and a mild effect of the protease mutation due to the presence of flexible structural elements at critical locations in the inhibitor molecule. The conclusions derived from the HIV-1 protease provide important thermodynamic guidelines that can be implemented in general drug design strategies. PMID- 11396920 TI - Molecular mechanisms of the resistance to hydrogen peroxide of enzymes involved in the calvin cycle from halotolerant Chlamydomonas sp. W80. AB - cDNA clones encoding NADP(+)-glyceraldehyde-3-phosphate dehydrogenase (NADP(+) GAPDH) and sedoheptulose-1,7-bisphosphatase (SBPase) were isolated and characterized from halotolerant Chlamydomonas sp. W80 (C. W80) cells. The cDNA clone for NADP(+)-GAPDH encoded 369 amino acid residues, preceded by the chloroplast transit peptide (37 amino acid residues). The cDNA clone for SBPase encoded 351 amino acids with the chloroplast transit peptide. The activities of NADP(+)-GAPDH and SBPase from C. W80 cells were resistant to H(2)O(2) up to 1 mM, as distinct from spinach chloroplastic thiol-modulated enzymes. The illumination to the dark-adapted cells and dithiothreitol treatment to the crude homogenate had little effect on the activities of NADP(+)-GAPDH and SBPase in C. W80. Modeling of the tertiary structures of NADP(+)-GAPDH and SBPase suggests that resistance of the enzymes to H(2)O(2) in C. W80 is due to the different conformational structures in the vicinity of the Cys residues of the chloroplastic enzymes between higher plant and C. W80 cells. PMID- 11396921 TI - Partial characterization of matrix components interacting with cartilage proteoglycans. AB - The charge content of aqueous suspensions of milled cartilage samples was determined by a colloid titration technique using a particle charge detector, and the data were compared with estimates from chemical analyses. Results indicated a close correlation between charge content determined by titration and that estimated by chemical analyses for samples of nasal septa only (a nonarticular cartilage). Such correlation did not hold for articular cartilages (metacarpalphalangeal joint and patella); extraction of these tissues with 0.1 or 1.2 M NaCl markedly increased the availability of the negative groups. Protein analysis, by SDS--PAGE, of the 1.2 M extracts indicated the presence of basic proteins, some of collagenous origin, such as chondrocalcin and proline-arginine rich protein, and some of noncollagenous proteins such as pleiotrophin and histone-H2b. These data thus suggest electrostatic interactions between these basic proteins and the negative groups of proteoglycans. Such interactions would have an important effect on the osmotic properties and in the organization of cartilage. PMID- 11396922 TI - Expression and degradation of the cystic fibrosis transmembrane conductance regulator in Saccharomyces cerevisiae. AB - Many cystic fibrosis disease-associated mutations cause a defect in the biosynthetic processing and trafficking of the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Yeast mutants, defective at various steps of the secretory pathway, have been used to dissect the mechanisms of biosynthetic processing and intracellular transport of several proteins. To exploit these yeast mutants, we have employed an expression system in which the CFTR gene is driven by the promoter of a structurally related yeast ABC protein, Pdr5p. Pulse-chase experiments revealed a turnover rate similar to that of nascent CFTR in mammalian cells. Immunofluorescence microscopy showed that most CFTR colocalized with the endoplasmic reticulum (ER) marker protein Kar2p and not with a vacuolar marker. Degradation was not influenced by the vacuolar protease mutants Pep4p and Prb1p but was sensitive to the proteasome inhibitor lactacystin beta-lactone. Blocking ER-to-Golgi transit with the sec18-1 mutant had little influence on turnover indicating that it occurred primarily in the ER compartment. Degradation was slowed in cells deficient in the ER degradation protein Der3p as well as the ubiquitin-conjugating enzymes Ubc6p and Ubc7p. Finally a mutation (sec61-2) in the translocon protein Sec61p that prevents retrotranslocation across the ER membrane also blocked degradation. These results indicate that whereas approximately 75% of nascent wild-type CFTR is degraded at the ER of mammalian cells virtually all of the protein meets this fate on heterologous expression in Saccharomyces cerevisiae. PMID- 11396923 TI - Site-directed mutagenesis of the heme axial ligands in the hemoflavoenzyme cellobiose dehydrogenase. AB - Cellobiose dehydrogenase (CDH) from Phanerochaete chrysosporium is an extracellular 90-kDa hemoflavoenzyme, organized into an N-terminal heme domain and a C-terminal flavin domain. The amino acid residues Met65 and His114 or His163 were suggested to be heme iron ligands. Mutations of these residues were made and mutant proteins were characterized. H114A mutant cultures produce a stable hemoflavoenzyme with spectral and kinetic characteristics similar to those of wild-type CDH. The M65A and H163A transformants secrete a 90-kDa hemoflavoenzyme, which oxidizes cellobiose in the presence of 2,6-dichlorophenol indophenol (DCPIP), but is unable to reduce cytochrome c. The heme domains of the M65A and H163A CDH variants are, however, unstable and susceptible to degradation, both yielding a 70-kDa cellobiose-oxidizing flavoenzyme. The spectral and kinetic characteristics of these truncated variants suggest that they contain only their respective flavin domains. The yield of the 90-kDa proteins was low and the proteins could not be purified to homogeneity; however, absorption spectra indicate that the 90-kDa proteins do contain the heme domain. Like the truncated flavoenzymes, the 90-kDa variants reduce DCPIP but are unable to transfer electrons to cytochrome c, in contrast to wild-type CDH. These findings suggest that H163 and M65 are the axial heme ligands and that both ligands are required for the reactivity and structural integrity of the heme domain. PMID- 11396924 TI - Reconstitution of the enzymatic activities of cytochrome P450s using recombinant flavocytochromes containing rat cytochrome b(5) fused to NADPH--cytochrome P450 reductase with various membrane-binding segments. AB - The role of the hydrophobic membrane-binding segments of NADPH-cytochrome P450 reductase (CPR) and cytochrome b(5) remain undefined. We have expressed four different recombinant flavocytochromes containing b(5) linked to CPR with different hydrophobic segments as linkers. These fusion proteins have been expressed in Escherichia coli and purified and some of their physical properties and electron transfer activities described in the accompanying paper. Of interest is the presence of internal "membrane-binding" hydrophobic segments in these flavocytochromes. This paper describes the ability of these flavocytochromes to reconstitute in vitro two P450 activities that have been reported to be stimulated by the addition of b(5) (the 17,20-lyase activity of CYP17A and the 6 beta hydroxylation of testosterone catalyzed by CYP3A4) and two P450 reactions that do not respond to the presence of b(5) (the 17 alpha-hydroxylation of progesterone catalyzed by CYP17A and the omega hydroxylation of lauric acid catalyzed by CYP4A1). The present study shows that a hydrophobic "membrane binding" segment must be present in the artificial flavocytochromes in order to successfully reconstitute in vitro hydroxylation activities with P450s. Differences in the effectiveness of the different flavocytochromes to reconstitute enzymatic activities depends on the P450 tested and the nature of the hydrophobic linker segment present in the purified recombinant flavocytochromes. The hypothesis is proposed that differences in the surface topology of a P450 may dictate differences in their docking with the CPR or b(5) component of a fusion protein, resulting in differences in the rates of electron transfer to the P450. PMID- 11396925 TI - Expression, purification, and physical properties of recombinant flavocytochrome fusion proteins containing rat cytochrome b(5) linked to NADPH-cytochrome P450 reductase by different membrane-binding segments. AB - Reconstitution of the enzymatic activities using purified microsomal cytochrome P450s (P450) requires the presence of a membrane-binding segment in the mammalian flavoprotein, NADPH--cytochrome P450 reductase (CPR), and the hemeprotein, cytochrome b(5) (b(5)). The mechanism(s) by which the membrane-binding segments of these proteins exert such a critical role in influencing the reconstitution of the NADPH-supported activity of a P450 remains undefined. In the present work we describe the construction, expression, and purification of four different types of recombinant flavocytochromes containing rat b(5) and rat CPR linked by various membrane-binding segments. The physical properties of these artificial fusion proteins have been studied to determine their ability to serve as electron transfer agents. These studies are a prelude to the subsequent study (accompanying paper) evaluating the functional roles of the hydrophobic (membrane binding) sequences of b(5) and CPR in the reconstitution of P450 activities. The present study shows that the purified recombinant fusion proteins can serve as active electron transport carriers from NADPH to cytochrome c as well as b(5) by intramolecular as well as intermolecular reactions. It is shown here that the electron transport properties of these purified fusion proteins are influenced by high concentrations of KCl, suggesting a role for charged amino acids in protein protein interactions. The present study illustrates the application of artificial recombinant flavocytochromes as useful proteins for the study of intramolecular electron transport reactions for comparison with intermolecular interactions. PMID- 11396926 TI - Proteoglycan depletion and magnetic resonance parameters of articular cartilage. AB - Calcium ions and various amounts of proteoglycans were removed from porcine articular cartilage explants using ethylenediaminetetraacetic acid or guanidinium chloride solutions. The water proton magnetic parameters such as T(1) and T(2) relaxation times, diffusion (D), and magnetization transfer (M(S)/M(0)) were then measured by 1D MR microscopy on native specimens, after incubation in the extracting solutions and after final reconditioning in a physiological saline. While the replacement of the interstitial fluid by the treating solutions strongly affected the various MR parameters, calcium depletion did not show any influence on the MRI appearance of the chondral tissue. Interestingly, only the longitudinal relaxation time T(1) and the diffusion coefficient D were seen to be sensitive to an extensive proteoglycan depletion of the tissue. Our results indicate that a modest proteoglycan depletion, as it occurs in the early stage of a pathological cartilage degradation, has little relevance to the above MR parameters. Further MRI studies on the macromolecular components of cartilage are, therefore, necessary for a better understanding of the interaction mechanisms between water and extracellular matrix that might lead to the early diagnosis of the cartilage damage. PMID- 11396927 TI - Substrate-bound fibronectin enhances scavenger receptor activity of macrophages by calcium signaling. AB - We have previously found that ability of mouse macrophages to bind and take up oxidized low-density lipoprotein (oxLDL) through scavenger receptors is significantly enhanced when the cells are plated on fibronectin (FN)-coated culture substrates. Here, the mechanisms of the enhancement of the scavenger receptor activity by the substrate-bound FN was investigated using thioglycollate induced mouse peritoneal macrophages. A Ca(2+) channel blocker diltiazem and a calmodulin inhibitor W-7 reduced the scavenger receptor activity of the macrophages plated on FN-coated substrate to the level of the cells plated on uncoated substrate, as assessed by oxLDL binding, while the scavenger receptor activity of the macrophages on uncoated substrate was little affected. Similarly, FN-induced enhancement of the scavenger receptor activity assessed by oxLDL uptake was selectively inhibited by Ca(2+) channel blockers (diltiazem, nifedipine, verapamil) and calmodulin inhibitors (W-7, trifluoperazine). Intracellular free Ca(2+) level of the macrophages was increased, depending on extracellular Ca(2+), when plated on FN-coated substrate. This increase in the Ca(2+) level was inhibited by diltiazem and RGD-containing peptides present in cell adhesive region of FN. Like the substrate-bound FN, Ca(2+) ionophore A23187 enhanced the scavenger receptor activity of binding and taking up of oxLDL. These results indicate that substrate-bound FN enhances scavenger receptor activity of macrophages by increasing channel-dependent Ca(2+) influx. A microtubule disruptor, colchicine, and an actin filament disruptor, cytochalasin B, inhibited the FN-induced enhancement of the scavenger receptor activity, suggesting that these cytoskeletal structures are required for transmission of the adhesion signal of FN. The number of the scavenger receptors was found to increase by 1.4 fold upon adhesion signal of FN. We suggest that substrate-bound FN increases the number of the macrophage scavenger receptors as a result of induction of Ca(2+) influx and causes increased accumulation of oxLDL within the cells, rendering the cells more susceptible to conversion into foam cells. PMID- 11396928 TI - Estrogen receptors alpha and beta exhibit different estradiol and estrogen response element binding in the presence of nonspecific DNA. AB - Estrogen receptors (ER) alpha and beta bind estradiol (E(2)) and estrogen response element (ERE) DNA sequences with high affinities. The different migration of ER--ERE complexes in the presence or absence of nonspecific DNA suggests that DNA may affect ER conformation and function. We measured the rate of E(2)--ER association and specific ER--ERE binding capacity (ERE--SBC) in the presence or absence of nonspecific DNA. Whereas DNA did not alter the rate of E(2)--ER alpha association, both ERE-containing and plasmid DNA decreased the rate of E(2) association with ER beta. Poly(dI-dC) decreased ERE--SBC of ER alpha, but did not affect the ERE--SBC of ER beta. Salmon sperm genomic DNA decreased the ERE--SBC of ER alpha, but increased the ERE--SBC of ER beta. We speculate that interaction of ER with genomic DNA may contribute to ER activation and play a role in the observed differences in transcriptional activity of ER alpha and ER beta. PMID- 11396929 TI - Taxol biosynthesis: differential transformations of taxadien-5 alpha-ol and its acetate ester by cytochrome P450 hydroxylases from Taxus suspension cells. AB - The biosynthesis of the diterpenoid antineoplastic drug Taxol in Taxus species involves the cyclization of the ubiquitous isoprenoid intermediate geranylgeranyl diphosphate to taxa-4(5),11(12)-diene followed by cytochrome P450-mediated hydroxylation (with allylic rearrangement) of this olefin precursor to taxa 4(20),11(12)-dien-5 alpha-ol, and further oxygenation and acylation reactions. Based on the abundances of naturally occurring taxoids, the subsequent order of oxygenation of the taxane core is considered to occur at C10, then C2 and C9, followed by C13, and finally C7 and C1. Circumstantial evidence suggests that the acetylation of taxadien-5 alpha-ol may constitute the third specific step of Taxol biosynthesis. To determine whether taxadienol or the corresponding acetate ester serves as the direct precursor of subsequent oxygenation reactions, microsomal preparations isolated from induced Taxus cells and optimized for cytochrome P450 catalysis were incubated with each potential substrate. Both taxadienol and taxadienyl acetate were oxygenated to the level of a diol and to higher polyols at comparable rates by cytochrome P450 enzymes of the microsomal preparation. Preparative-scale incubation allowed the isolation of sufficient quantities of the diol derived from taxadienol to permit the NMR-based structural elucidation of this metabolite as taxa-4(20),11(12)-dien-5 alpha,13 alpha-diol, which may represent an alternate route of taxoid metabolism in induced cells. GC MS-based structural definition of the diol monoacetate derived in microsomes from taxadienyl acetate confirmed this metabolite as taxa-4(20),11(12)-dien-5 alpha acetoxy-10 beta-ol, thereby indicating that acetylation at C5 of taxadienol precedes the cytochrome P450-mediated insertion of the C10-beta-hydroxyl group of Taxol. PMID- 11396930 TI - Demonstration that menthofuran synthase of mint (Mentha) is a cytochrome P450 monooxygenase: cloning, functional expression, and characterization of the responsible gene. AB - (+)-Menthofuran is an undesirable monoterpenoid component of peppermint (Mentha x piperita) essential oil that is derived from the alpha,beta-unsaturated ketone (+)-pulegone. Microsomal preparations, from the oil gland secretory cells of a high (+)-menthofuran-producing chemotype of Mentha pulegium, transform (+) pulegone to (+)-menthofuran in the presence of NADPH and molecular oxygen, implying that menthofuran is synthesized by a mechanism analogous to that of mammalian liver cytochrome P450s involving the hydroxylation of the syn-methyl group of (+)-pulegone, spontaneous intramolecular cyclization to the hemiketal, and dehydration to the furan. An abundant cytochrome P450 clone from a peppermint oil gland cell cDNA library was functionally expressed in Saccharomyces cerevisiae and Escherichia coli and shown to encode the (+)-menthofuran synthase (i.e., (+)-pulegone-9-hydroxylase). The full-length cDNA contains 1479 nucleotides, and encodes a protein of 493 amino acid residues of molecular weight 55,360, which bears all of the anticipated primary structural elements of a cytochrome P450 and most closely resembles (35% identity) a cytochrome P450 monoterpene hydroxylase, (+)-limonene-3-hydroxylase, from the same source. The availability of this gene permits transgenic manipulation of peppermint to improve the quality of the derived essential oil. PMID- 11396931 TI - Nitric oxide from the inducible nitric oxide synthase (iNOS) increases the expression of cytochrome P450 2E1 in iNOS-null hepatocytes in the absence of inflammatory stimuli. AB - Nitric oxide (NO) can modulate numerous genes through several pathways, yet some genes may be modulated only in the presence of the inflammatory stimuli that upregulate the inducible nitric oxide synthase (iNOS) rather than by NO alone. Furthermore, the role of prior expression of iNOS in the modulation of genes by NO is unknown. We addressed these issues in hepatocytes harvested from iNOS-null (iNOS(-/-)) mice exposed to NO by treatment with NO donors or by infection with an adenovirus-expressing human iNOS (Ad-iNOS), rather than by stimulation with inflammatory cytokines. Differential display and gene array analyses performed on mRNA derived from iNOS(-/-) hepatocytes demonstrated that infection with Ad-iNOS, but not infection with a control adenovirus expressing the beta-galactosidase gene (Ad-LacZ), induced a gene fragment identical to cytochrome P450 2E1 (CYP2E1). Northern analysis performed with this fragment demonstrated that treatment of iNOS(-/-) hepatocytes with Ad-iNOS or with the NO donor S-nitroso-N acetyl-d,l-penicillamine (SNAP), but not control treatment or infection with Ad LacZ, resulted in increased expression of CYP2E1. Inhibition of soluble guanylyl cyclase partially blocked the induction of CYP2E1 mRNA by Ad-iNOS. Rat hepatocytes treated with SNAP also exhibited increased expression of CYP2E1 mRNA. Preliminary studies, however, suggest that the induction of CYP2E1 in the rat hepatocytes treated with cytokines was not reduced in the presence of a NOS inhibitor. Our results suggest that CYP2E1 can be induced solely by NO derived from iNOS, at least partly in a cyclic GMP-dependent manner and independently of inflammatory stimuli or of prior exposure to NO. PMID- 11396932 TI - The Membrane-bound L- and D-lactate dehydrogenase activities in mitochondria from Euglena gracilis. AB - The activity of the pyridine nucleotide-independent lactate dehydrogenase (iLDH) was characterized in mitochondria isolated from the protist Euglena gracilis. The dissociation constants for L- and D-lactate were similar, but the V(max) was higher with the d isomer. A ping-pong kinetic mechanism was displayed with 2,4 dichlorophenol-indolphenol (DCPIP), or coenzyme Q(1), reacting as the second substrate with the modified, reduced enzyme. Oxamate was a competitive inhibitor against both L- and D-lactate. Oxalate exerted a mixed-type inhibition regarding L- or D-lactate and also against DCPIP. The rate of L-lactate uptake was partially inhibited by mersalyl and lower than the rate of dehydrogenation, which was mersalyl-insensitive. These data suggested that the active site of L-iLDH was orientated toward the intermembrane space. The following observations indicated the existence of two stereo-specific iLDH enzymes in the inner membrane of Euglena mitochondria: a greater affinity of the D-iLDH for both inhibitors, D iLDH thermo-stability at 70 degrees C and denaturation of L-iLDH, opposite signs in the enthalpy change for the association reaction of the isomers to the enzyme, differential solubilization of both activities with detergents, and different molecular mass. PMID- 11396933 TI - Formation of compound I and compound II ferryl species in the reaction of hemoglobin I from Lucina pectinata with hydrogen peroxide. AB - The formation of ferryl heme (Fe(IV) = O) species, i.e., compound I and compound II, has been identified as the main intermediates in heme protein peroxidative reactions. We report stopped-flow kinetic measurements which illustrate that the reaction of hemoglobin I (HbI) from Lucina pectinata with hydrogen peroxide produce ferryl intermediates compound I and compound II. Compound I appears relatively stable displaying an absorption at 648 nm. The rate constant value (k'(2)) for the conversion of compound I to compound II is 3.0 x 10(-2) s(-1), more than 100 times smaller than that reported for myoglobin. The rate constant value for the oxidation of the ferric heme (k'(12) + k'(13)) is 2.0 x 10(2) M(-1) s(-1). These values suggest an alternate route for the formation of compound II (by k'(13)) avoiding the step from compound I to compound II (k'(2)). In HbI from L. pectinata the stabilization of compound I is attribute to the unusual collection of amino acids residues (Q64, F29, F43, F68) in the heme pocket active site of the protein. PMID- 11396934 TI - Histone H1: an antimicrobial protein of Atlantic salmon (Salmo salar). AB - Antimicrobial activity was detected in acid extracts of liver, intestine, and stomach of healthy Atlantic salmon (Salmo salar). An antimicrobial protein was isolated from salmon liver using acid extraction followed by ammonium sulfate precipitation, large-scale gel filtration chromatography, reverse-phase HPLC, and size exclusion HPLC. The salmon antimicrobial (SAM) protein was found to have a molecular mass of 20,734 Da by MALDI TOF mass spectrometry. Peptide mass fingerprinting and partial sequencing by tandem nanoelectrospray mass spectrometry identified the protein as histone H1. The protein had a minimal inhibitory concentration of 31 microg/mL against E. coli D31 in a plate clearing assay. The effect of the SAM protein on bacterial morphology was indistinguishable from that of (Ala-(8,13,18))-magainin II, as shown by scanning electron microscopy, which suggests that the protein disrupts E. coli membranes in a manner similar to that of most antimicrobial peptides. This protein may act as an antimicrobial in vivo through active secretion or by release from cells during infection-related apoptosis. PMID- 11396935 TI - Processing of DNase domain during translocation of colicin E7 across the membrane of Escherichia coli. AB - Translocation of colicin across the membrane of sensitive cells has been studied extensively. However, processing of the toxicity domain of colicin during translocation has been the subject of much controversy. To investigate the final translocation product of colicin across the membrane of Escherichia coli, an endogenously expressed His-tagged Im7 protein was constructed to detect any translocation product containing the DNase domain traversed the inner membrane into cytoplasm of the E. coli cells. As a result, a final processed DNase domain of ColE7 was identified in the intracellular space of the cells treated with Col Im complex. In the presence of periplasmic extracts, in vitro processing of DNase domain of ColE7 was also observed. These results suggest that the processing of ColE7 has occurred for translocation of the DNase-type colicin across the membrane and the process is probably taking place in the periplasmic space of the membrane. PMID- 11396936 TI - Iron extraction from soybean lipoxygenase 3 and reconstitution of catalytic activity from the apoenzyme. AB - Lipoxygenases contain a unique nonheme iron cofactor with a redox role in the catalyzed reaction. The conditions for the extraction of the metal atom were investigated for one of the soybean lipoxygenase isoenzymes. Removal of the iron by o-phenanthroline was attained in the presence of substrate under anaerobic conditions, but the apoenzyme could not be isolated and reconstituted. The freshly regenerated sodium form of Chelex-100 also removes the iron atom from native soybean lipoxygenase 3, but only in sodium bicarbonate buffer at pH 8.0. The soluble but inactive apoenzyme was reconstituted with ferric ammonium sulfate in Tris--HCl buffer at pH 7.0. Stoichiometric iron in the reconstituted enzyme was established using inductively coupled plasma-atomic emission spectroscopy. The reconstituted enzyme contained 90 +/- 10% of the specific activity of the native enzyme. The native configuration of the reconstituted iron site was confirmed by electron paramagnetic resonance spectroscopy. PMID- 11396937 TI - Effect of pH on stability of anthrax lethal factor: correlation between denaturation and activity. AB - Anthrax is caused by Gram positive bacterium Bacillus anthracis. Pathogenesis is result of production of three protein components, protective antigen (PA), lethal factor (LF), and edema factor (EF). PA in combination with LF (lethal toxin) is lethal to animals, while PA in combination with EF (edema toxin), causes edema. PA, LF, and EF are very thermolabile. Differential scanning calorimetry (DSC) was used to unravel the energetics of LF denaturation as a function of pH ranging from 7.8 to 5.5. Transition temperature (T(m)) of LF was found to be approximately equal to 42 degrees C and onset of denaturation occurs at approximately equal to 30 degrees C. The ratio of calorimetric to van't Hoff's enthalpy was nearly equal to unity at pH 7.0, indicative of presence of single structural domain in LF at pH 7.0, unlike PA which has been structurally observed to consist of 4 domains. It was found by cytotoxicity studies using J774A.1 macrophage like cells that LF was most stable at pH approximately 6.5. This paper reports for the first time the denaturation of LF at different pH values at 37 degrees C and tries to establish a correlation between denaturation and loss of LF activity at different pH values. PMID- 11396938 TI - A stress-responsive gene from groundnut, Gdi-15, is homologous to flavonol 3-O glucosyltransferase involved in anthocyanin biosynthesis. AB - Stress-tolerant crops are expected to express genes not normally expressed in susceptible crops. We have used desiccation stress coupled with high light intensity to identify groundnut as a relatively tolerant crop. Stress-responsive genes (Gdi, Groundnut Desiccation Induced) were cloned by subtractive hybridisation. The sequence of Gdi-15 shows homology to flavonol 3-O glucosyltransferases, which are involved in anthocyanin biosynthesis. Gdi-15 transcripts increase markedly in response to stress, suggestive of a role in stress tolerance. PMID- 11396939 TI - Novel platelet antiaggregating substances. AB - We describe herein a novel action of four stable analogs of the hepoxilins. These analogs inhibit to different degrees, the aggregation of washed human platelets evoked by collagen. One of the analogs, PBT-3, is particularly effective with an IC(50) = 8 x 10(-7) M. The other analogs are about 5-fold less active, but all analogs are about 500-fold more active than the native hepoxilins. The PBT analogs inhibit the collagen-enhanced formation of thromboxane A(2) and HHT but do not affect the formation of 12-HETE or the release of arachidonic acid except at doses higher than those needed to block platelet aggregation. These results demonstrate that these novel compounds may have potential for development into drugs in the treatment of thromboxane-mediated cardiovascular disease. PMID- 11396940 TI - Interferon-alpha promotes survival of human primary B-lymphocytes via phosphatidylinositol 3-kinase. AB - Signaling pathways for the antiviral and antiproliferative biological effects of type I interferons (IFN) are well established. In this report we demonstrate a novel signaling pathway for IFN-alpha, as it induced rapid phosphorylation of both PKB/Akt and its substrate forkhead. The PI3-kinase inhibitor LY294002 abolished these phosphorylations. PI3-kinase has been implicated in cell survival mediating its effect through the second messenger PIP3 and the subsequent activation of PKB/Akt. We could show that IFN-alpha inhibited spontaneous apoptosis of primary B-lymphocytes, in the absence of a mitogenic stimulus. This effect was inhibited by LY294002. Thus, our data suggests that IFN-alpha promotes survival of peripheral B-lymphocytes via the PI3-kinase-PKB/Akt pathway. In addition, IFN-alpha stimulation of anti-IgM activated cells resulted in downregulated expression of the cell cycle inhibitor p27/Kip1. PMID- 11396941 TI - Riboflavin synthesis genes ribE, ribB, ribH, ribA reside in the lux operon of Photobacterium leiognathi. AB - Nucleotide sequence of the riboflavin synthesis genes ribE, ribB, ribH, ribA (GenBank Accession No. AF364106) resided in the lux operon of Photobacterium leiognathi PL741 has been determined, and the amino acid sequences of riboflavin synthetase (RibE), DHBP synthetase (RibB), lumazine synthetase (RibH), GTP cyclohydrolase II (RibA) encoded by the riboflavin synthesis genes are deduced. Nucleotide sequence reveals that the ribE gene encodes the riboflavin synthetase responsible for converting lumazine to riboflavin, the ribB gene encodes the DHBP synthetase responsible for 3,4-dihydroxyl-2-butanone 4-phosphate synthesis, the ribH gene encodes the lumazine synthetase responsible for lumazine synthesis, and the ribA gene encodes the GTP cyclohydrolase II responsible for lumazine synthesis. Functional analysis illustrates that the specific segments lay behind the ribH and ribA genes might form potential loops Omega(oT) and Omega(TI)- Omega(TII); Omega(oT) is functioned as mRNA stability loop or/and for subregulation by alternative modulation, and Omega(TI)--Omega(TII) could be the transcriptional terminator of the lux operon. The gene order of the ribE, ribB, ribH, ribA genes resided in the lux operon and linked to the lum operon is <- ter*-lumQ-lumP-R&R-luxC-luxD-luxA-luxB-luxN-luxE-luxG-ribE-ribB-ribH-ribA-ter--> (R&R: regulatory region; ter: transcriptional terminator), whereas the R&R is the regulatory region for the lum and the lux operons, and ter and ter* are the transcriptional terminators for the lux and lum operons. PMID- 11396942 TI - Constitutive activation of A(3) adenosine receptors by site-directed mutagenesis. AB - The objective of this study was to create constitutively active mutant human A(3) adenosine receptors (ARs) using single amino acid replacements, based on findings from other G protein-coupled receptors. A(3) ARs mutated in transmembrane helical domains (TMs) 1, 3, 6, and 7 were expressed in COS-7 cells and subjected to agonist radioligand binding and phospholipase C (PLC) and adenylyl cyclase (AC) assays. Three mutant receptors, A229E in TM6 and R108A and R108K in the DRY motif of TM3, were found to be constitutively active in both functional assays. The potency of the A(3) agonist Cl-IB-MECA (1-chloro-N(6)-(3-iodobenzyl)adenosine-5' N-methyluronamide) in PLC activation was enhanced by at least an order of magnitude over wild type (EC(50) 951 nM) in R108A and A229E mutant receptors. Cl IB-MECA was much less potent (>10-fold) in C88F, Y109F, and Y282F and mutants or inactive following double mutation of the DRY motif. The degree of constitutive activation was more pronounced for the AC signaling pathway than for the PLC signaling pathway. The results indicated that specific locations within the TMs proximal to the cytosolic region were responsible for constraining the receptor in a G protein-uncoupled conformation. PMID- 11396943 TI - Identification of calnexin as a binding protein for Amadori-modified glycated albumin. AB - Albumin modified by Amadori glucose adducts (glycated albumin) selectively binds to glomerular mesangial cells and triggers signal transduction processes that modulate cellular function. To identify glycated albumin binding proteins, we applied membrane extracts prepared from murine mesangial cells to a column of lysine-Sepharose followed by application to an affinity column of fructosyllysine Sepharose. This procedure yielded an approximately 90 kDa polypeptide that immunoreacted with Amadori-modified but not carbohydrate-free albumin. MALDI mass fingerprinting matched 9 out of 25 peptides with calnexin, and amino acid analysis showed homology with this transmembrane calcium-binding protein of the calreticulin family. These results indicate that one of the mesangial cell receptors for glycated albumin is a calnexin-like protein. PMID- 11396944 TI - Serum after partial hepatectomy stimulates iNOS gene transcription via downstream NF-kappa B site. AB - It has been known that the expression of inducible nitric oxide synthase (iNOS) is up-regulated during hepatic regeneration. The present study characterized the molecular mechanisms involved in the transcriptional activation of iNOS gene by using the serum after partial hepatectomy (post-PH serum) in vitro. The post-PH serum rapidly induced iNOS mRNA expression, which was blocked by anti-tumor necrosis factor-alpha (TNF-alpha) antibody in BNL CL.2 cells, murine embryonic liver cell line. In addition, EMSAs using a NF-kappa B-specific oligomer showed that the up-regulated iNOS mRNA expression in cells treated with post-PH serum correlated with transient activation of NF-kappa B complex (p50/p65 heterodimer). Transient transfection of BNL CL.2 cells with iNOS promoter linked to a CAT reporter gene showed the transcriptional activation of iNOS promoter by post-PH serum. Furthermore, site-directed mutational analysis of the two NF-kappa B sites individually or in combination revealed that iNOS expression by post-PH serum is regulated by the downstream NF-kappa B site, but not by upstream NF-kappa B site. Taken together, these results suggest that the downstream NF-kappa B site acts as an essential component for the iNOS expression by post-PH serum during hepatic regeneration. PMID- 11396945 TI - A yeast two-hybrid study on self-association of the ORF2 protein of hepatitis E virus. AB - Hepatitis E virus is a human RNA virus containing three open reading frames. Of these, ORF2 encodes the major capsid protein (pORF2) and may possess regulatory functions, in addition to a structural one. In this study, we have shown using the yeast two-hybrid system and in vitro immobilization experiments that full length pORF2 is capable of self-association, thus forming a homodimer. Using mutational analysis we have studied dimerization of various truncated versions of the ORF2 capsid protein using the yeast two-hybrid system and supported our findings with in vitro immobilization experiments. Deletions of pORF2 reveal a loss of the dimerization potential for all deletions except an N-terminal 127 amino-acid deletion. Our studies suggest that the dimerization property of pORF2 may not be amino-acid sequence dependent but instead a complex formation of a specific tertiary structure that imparts pORF2 its property to self-associate. PMID- 11396946 TI - MAPK and SRC-kinases control EGR-1 and NF-kappa B inductions by changes in mechanical environment in osteoblasts. AB - Bone loss occurs in microgravity whereas an increase in bone mass is observed after skeletal loading. This tissue adaptation involves changes in osteoblastic proliferation and differentiation whose mechanisms remain largely unknown. In this context, we investigated the expression and the nuclear translocation of Egr 1 and NF-kappa B, in a simulated microgravity model (clinostat) and in a model of mechanical strain (Flexcell). We performed RT-PCR and immunocytochemistry analyses at baseline and up to 2 h after stimulation (a mitogenic regimen, 1% stretch, 0.05 Hz, 10 min, or clinorotation 50 rpm, 10 min) in osteoblastic ROS17/2.8 cells. Egr-1 induction as well as NF-kappa B nuclear translocation were activated by mechanical changes. PKC downregulation and COX1/2 inhibition did not alter these inductions. In contrast, ERK1/2, p38(MAPK) and src-kinases pathways were differentially involved in both models. Thus, we demonstrated that changes in the mechanical environment induced an activation of Egr-1 and NF-kappa B with specific kinetics and involved various transduction pathways including MAPKs and src-kinases. These could partially explain the later alterations of proliferation observed. PMID- 11396947 TI - (31)P NMR of apicomplexans and the effects of risedronate on Cryptosporidium parvum growth. AB - High-resolution 303.6 MHz (31)P NMR spectra have been obtained of perchloric acid extracts of Plasmodium berghei trophozoites, Toxoplasma gondii tachyzoites, and Cryptosporidium parvum oocysts. Essentially complete resonance assignments have been made based on chemical shifts and by coaddition of authentic reference compounds. Signals corresponding to inorganic pyrophosphate were detected in all three species. In T. gondii and C. parvum, additional resonances were observed corresponding to linear triphosphate as well as longer chain polyphosphates. Spectra of P. berghei and T. gondii also indicated the presence of phosphomonoesters and nucleotide phosphates. We also report that the pyrophosphate analog drug, risedronate (used in bone resorption therapy), inhibits the growth of C. parvum in a mouse xenograft model. When taken together, our results indicate that all the major disease-causing apicomplexan parasites contain extensive stores of condensed phosphates and that as with Plasmodium falciparum and T. gondii, the pyrophosphate analog drug risedronate is an inhibitor of C. parvum cell growth. PMID- 11396948 TI - Differential effects of primary human immunodeficiency virus type 1 nef sequences on downregulation of CD4 and MHC class I. AB - We analyzed two primary nef sequences, KS2 (subtype B) and K306 (subtype D), each directly isolated from patients. Cell lines constitutively expressing respective Nef proteins were constructed using a retroviral vector. There were significant differences in the ability to downregulate surface CD4 and MHC class I proteins between different nef sequences. When the nef sequence from NL432 was used as a reference, KS2 Nef demonstrated the highest ability to downregulate MHC class I, whereas it appeared to lack the ability to downregulate CD4. On the contrary, Nef from K306 decreased the level of surface CD4 to a greater extent, but was less effective on downregulation of MHC class I. These results showed that the levels of downregulation of CD4 and MHC class I could significantly vary among HIV strains and that two well-known functions of Nef, downregulation of CD4 and MHC class I, would be separated. PMID- 11396949 TI - Isolation of a novel human gene, ARHGAP9, encoding a rho-GTPase activating protein. AB - Members of the Rho family of small guanosine triphosphatases (Rho-GTPases) have emerged as key coordinators of signaling pathways leading to remodeling of the actin cytoskeleton, a process that plays a critical role in cell adhesion and migration. However, the precise regulatory mechanisms remain to be elucidated. Here we report isolation of a novel human gene, ARHGAP9, which encodes a protein containing a Rho-GTPase activating protein (Rho-GAP) domain, a src-homology 3 (SH3) domain, a pleckstrin homology (PH) region, and a WW domain. In vitro, the recombinant protein revealed substantial GAP activity toward Cdc42Hs and Rac1, and less toward RhoA. The transcript was predominantly expressed in peripheral blood leukocytes, spleen, and thymus. Exogenous expression of the entire coding region of ARHGAP9 into human leukemia KG-1 cells repressed adhesion of the cells to fibronectin and collagen IV. Our results indicate that ARHGAP9 is involved in regulating adhesion of hematopoietic cells to extracellular matrix. PMID- 11396950 TI - A new locus for a dominant form of multinodular goiter on 3q26.1-q26.3. AB - A mass screening program for congenital hypothyroidism has markedly improved prognosis of children with congenital hypothyroidism and also revealed several cases with unknown pathogenesis. We here report two independent Japanese multigeneration families with multinodular goiter (MNG) with euthyroidism and with high TSH. The propositi, 3- and 8-year-old girls in two families, were found during a mass screening. An autosomal dominant pattern of inheritance was suggested in both families. The clinical examinations suggested impaired hormonogenesis but discarded known defects in iodine transport, organification, deficiency of hydrogen peroxide, and thyroid peroxidase. Linkage analysis of the two families including 10 members each using 343 microsatellite markers mapped a single locus independently at D3S1618 (theta = 0) on 3q26.1-q26.3 with a two point LOD score 3.62 (1.81 for each family) and multipoint LOD score of 3.61 (1.80 for each family). Haplotype inspection delimited an 18-cM interval between D3S1565 and D3S3686. PMID- 11396951 TI - Structural similarity of ghrelin derivatives to peptidyl growth hormone secretagogues. AB - Ghrelin is a 28-amino acid residue endogenous growth hormone secretagogue. Intensive investigations revealed that the N-terminus tetrapeptide, having octanoyl group at Ser(3), is the minimum active core. In this study, we further explored the structure-function relationships of the active N-terminus portion of ghrelin using a Ca(2+) mobilization assay. The smallest and most potent ghrelin derivative we have found so far is 5-aminopentanoyl-Ser(Octyl)-Phe-Leu aminoethylamide, showing comparable activity to the natural molecule. In the process of modifying the active core, the ghrelin-derived short analogues emerged structurally close to peptidyl growth hormone secretagogues. The N-terminus modification suggested that Gly(1)-Ser(2) unit works as a spacer, forming adequate distance between N(alpha)-amino group and n-octanoyl group. Replacement of 3rd and 4th amino acid residues to D-isomer suggested that the N-terminal dipeptide contributes to shape the biologically active geometry by effecting conformation of residues in positions 3 and 4. PMID- 11396952 TI - Hormonal regulation of the human ghrelin receptor gene transcription. AB - The aim of this study is to clarify the hormonal regulation of the human ghrelin receptor gene expression in GH(3) cells transfected with our previously cloned 5' flanking region inserted into a luciferase reporter vector. Phorbor 12 tetradecanoate 13-acetate (TPA) with simultaneous addition of Bay K8644 mimicking ghrelin action caused a significant inhibition of the luciferase activity through the ghrelin receptor gene upstream proximal to -669 but not to -608 base pairs (bp). Glucocorticoid caused a weak but significant inhibition of the luciferase activity through the ghrelin receptor gene upstream proximal to -531 but not to 475 bp. Electrophoretic mobility shift assay resulted in binding of oligonucleotides between -669 and -640 bp, and between -520 and -491 bp to GH(3) cell nuclear proteins unlike AP(2) or glucocorticoid receptor. These results suggest that both TPA/Bay K8644 and glucocorticoid downregulate human ghrelin receptor gene expression through the transcriptional mechanism involving some nuclear factors. PMID- 11396953 TI - Antioxidant enzyme peroxiredoxin 5 is upregulated in degenerative human tendon. AB - Peroxiredoxin 5 (PRDX5) is a novel thioredoxin peroxidase recently identified in a variety of human cells and tissues, which is considered to play an important role in oxidative stress protection mechanisms. However, little is known about its expression in tendon degeneration, a common and disabling condition that primarily affects older people, in which oxidative stress may be implicated. The present study demonstrated that normal human tendon expresses PRDX5 and its expression is significantly increased in degenerative tendon. In addition, we have localized PRDX5 to fibroblasts in normal tendon and to both fibroblasts and endothelial cells in degenerate tendon. The differential expression of PRDX5 in normal and degenerate tendon shows that a thioredoxin peroxidase with antioxidant properties is upregulated under pathophysiological conditions and suggests that oxidative stress may be involved in the pathogenesis of tendon degeneration. PRDX5 may play a protective role against oxidative stress during this pathophysiological process. PMID- 11396954 TI - Neuronal development of embryonic stem cells: a model of GABAergic neuron differentiation. AB - Neural cultures derived from differentiating embryonic stem (ES) cells are a potentially powerful in vitro model of neural development. We show that neural cells derived from mouse ES cells express mRNAs characteristic of GABAergic neurons. The glutamate decarboxylase genes (Gad1 and Gad2), required for GABA synthesis and the vesicular inhibitory amino acid transporter (Viaat) gene, required for GABA vesicular packaging are activated in the ES-derived cultures. Nearly half of the ES-derived neurons express the GAD67 protein, the product of the Gad1 gene. Building on these results we show that Gad1-lacZ "knockin" reporter ES cell lines can be used to easily monitor Gad1 expression patterns and expression levels during ES differentiation. We also demonstrate that the ES derived neural progenitors can be infected with retroviruses or transfected with plasmids via lipofection. These experiments outline the basic strategies and methods required for studies of GABAergic gene expression and regulation in ES derived neuronal cultures. PMID- 11396955 TI - Anti-atherogenic effect of citrus flavonoids, naringin and naringenin, associated with hepatic ACAT and aortic VCAM-1 and MCP-1 in high cholesterol-fed rabbits. AB - The anti-atherogenic effects of the citrus flavonoids, naringin and naringenin, were evaluated in high cholesterol-fed rabbits. At 3 months of age, 30 male New Zealand White (NZW) rabbits were divided into three groups (n = 10 per group). The rabbits were fed a 1% cholesterol diet alone (control group) or a diet supplemented with either 0.1% naringin or 0.05% naringenin for 8 weeks. The plasma lipoprotein levels, total cholesterol, triglyceride, and high-density lipoprotein showed no significant differences in the control and experimental groups. Hepatic acyl-CoA:cholesterol acyltransferase (ACAT) activity was slightly low in naringin (5.0%)- and naringenin (15.0%)-fed rabbits, compared to control group. The aortic fatty streak areas were significantly lower in both the naringin (19.2 +/- 5.6%)- and naringenin (18.1 +/- 6.5%)-supplemented groups than in the control group (60.4 +/- 14.0%). The expression levels of vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemotactic protein-1 (MCP-1), by semiquantitative RT-PCR analysis of the thoracic aorta, were significantly lower in the flavonoids supplemented groups than in the control group. These results suggest that the anti-atherogenic effect of the citrus flavonoids, naringin and naringenin, is involved with a decreased hepatic ACAT activity and with the downregulation of VCAM-1 and MCP-1 gene expression. PMID- 11396956 TI - Regulation of stearoyl-CoA desaturase activity by the trans-10,cis-12 isomer of conjugated linoleic acid in HepG2 cells. AB - Stearoyl-CoA desaturase (SCD) catalyzes the rate-limiting step in the cellular synthesis of monounsaturated fatty acids mainly oleate (C18:1) and palmitoleate (C16:1) which are the major monounsaturated fatty acids of membrane phospholipids, cholesterol esters, waxes, and triglycerides. Several SCD isoforms exist in the mouse whereas the human has one well-characterized SCD gene. The trans-10,cis-12 isomer of conjugated linoleic acid has been previously shown to repress the expression of the mouse SCD1 gene isomer by decreasing SCD gene expression as well as by direct inhibition of SCD enzyme activity. We studied the regulation of human stearoyl-CoA desaturase (SCD) expression by conjugated linoleic acid (CLA) in cultured human hepatoblastoma cell line, HepG2. Treatment of the cells with the trans-10,cis-12 CLA isomer did not cause changes in the SCD gene transcription, mRNA and protein levels. However, this isomer decreased both the SCD activity as well as the levels of monounsaturated fatty acids. The other major CLA isomer, cis-9,trans-11 CLA, had no effect on SCD gene expression and activity. These results suggest that in HepG2 cells the trans-10,cis-12 CLA isomer regulates human SCD activity mainly by a posttranslational mechanism. PMID- 11396957 TI - Differential amplification of hypervariable region 1 of hepatitis C virus by partially mismatched primers. AB - The quasispecies nature of hepatitis C virus (HCV) has been well documented over its whole genome and the most variable domain is located at the 5' end of the second envelope region, the so-called hypervariable region 1 (HVR1). HVR1 has therefore been extensively used as the target for characterizing HCV quasispecies profiles. In this study, we reported our finding that partially mismatched primers preferentially amplify different HVR1 sequences in a heterogeneous virus population. This finding suggests a possible mechanism of bias during the amplification of HVR1 sequences and may be responsible for some conflicting data regarding evolutionary or clinical implications of HCV quasispecies. PMID- 11396958 TI - Translocation of S100A1(1) calcium binding protein during heart surgery. AB - Myocardial ischemia during cardiopulmonary bypass terminated by reperfusion generally leads to different degrees of damage of the cardiomyocytes induced by transient cytosolic Ca(2+) overload. Recently, much attention has been paid to the role of heart-specific Ca(2+)-binding proteins in the pathogenesis of myocardial ischemia-reperfusion injury. S100A1 is a heart-specific EF-hand Ca(2+) binding protein that is directly involved in a variety of Ca(2+)-mediated functions in myocytes. The aim of our study was to investigate the localization and translocation of S100A1 in the human heart under normal (baseline) conditions and after prolonged ischemia and reperfusion of the myocardium. Our data suggest that S100A1 is directly involved in the transient perioperative myocardial damage caused by ischemia during open heart surgery in humans. Given its role in the contractile function of muscle cells, this S100 protein could be an important "intracellular link" in ischemia-reperfusion injury of the heart. PMID- 11396959 TI - Degradation of bisphenol A by purified laccase from Trametes villosa. AB - Degradation of bisphenol A (BPA), an endocrine-disrupting chemical, was studied with a purified laccase from the basidiomycete Trametes villosa. SDS- polyacrylamide gel electrophoresis of the purified laccase gave one single band with a mobility corresponding to MW 65 kDa. The absorption spectrum showed the characteristics of a blue copper protein with a maximum peak at 600 nm. HPLC analysis revealed that 2.2 micromol BPA were degraded by incubation with 1.5 units of the purified laccase in a total volume of 1.0 ml at pH 6.0 and 60 degrees C for 3 h. The enzyme reaction proceeded rapidly without requirement of mediators for the electron transfer. Isolation and identification of several reaction products are in progress, in which one product was identified as 4 isopropenylphenol by a gas chromatography--mass spectrophotometer. PMID- 11396960 TI - Advanced glomerulosclerosis is reversible in nephrotic mice. AB - Advanced glomerulosclerosis, a common hallmark of chronic renal diseases (CRD) is believed to be irreversible, and it is thought that glomerular hyperfiltration and hypertrophy may participate in its pathogenesis. We demonstrate here that glomerulosclerosis is "reversible" in an animal model. We used nephrotic ICGN (nep/nep) mice which showed a rapid progression of glomerulosclerosis, accompanied by histological findings for glomerular hyperfiltration. It is known that ureter ligation reduces glomerular filtration in ligated kidneys. When ureter ligation was applied to our model, glomerulosclerosis (characterized by myofibroblast hyperplasia and over-accumulated matrix protein) weakened in conjunction with suppressed glomerular hypertrophy. During this process, glomerular myofibroblasts showed apoptotic cell death after unilateral ureter ligation (UUO) treatment. Our results suggest that inhibition of glomerular filtration in sclerotic tufts may cause glomerular remodeling through the modulation of molecular and cellular sclerogenesis. PMID- 11396961 TI - A hematopoietic-specific transmembrane protein, Art-1, is possibly regulated by AML1. AB - The functions of AML1 in hematopoietic differentiation are repressed by AML1 mutants including the AML1/ETO chimeric protein, which is seen in t(8;21) acute myeloid leukemia. Erythroid progenitors of the patients with t(8;21) AML expressed AML1/ETO. To investigate the effect of AML1/ETO in erythroid cells, we made a tetracycline-regulated AML1/ETO overexpression system in mouse erythroleukemic (MEL) cells. Enforced AML1/ETO repressed the terminal erythroid differentiation. Furthermore, we performed representational difference analysis using this MEL cell system to clone the downstream targets of AML1 in erythroid cell differentiation. We cloned a novel transmembrane protein, Art-1 (AML1 regulated transmembrane protein 1), which is a member of tetramembrane spanning superfamily. Art-1 expression was restricted in hematopoietic cells. It was upregulated by AML1 and downregulated by AML1/ETO in both erythroid and myeloid cells, and increased during erythroid cell differentiation. Art-1 may play an important role in the differentiation of erythroid cells, possibly as a direct downstream target of AML1. PMID- 11396962 TI - Engineering of a manganese-binding site in lignin peroxidase isozyme H8 from Phanerochaete chrysosporium. AB - A Mn(2+)-binding site was created in the recombinant lignin peroxidase isozyme H8 from Phanerochaete chrysosporium. In fungal Mn peroxidase, the Mn-binding site is composed of Glu35, Glu39, and Asp179. We generated a similar site in lignin peroxidase by generating an anionic binding site. We generated three mutations: Asn182Asp, Asp183Lys, and Ala36Glu. Its activity, veratryl alcohol, and Mn(2+) oxidation were compared to those of native recombinant enzyme and to fungal Mn peroxidase isozyme H4, respectively. The mutated enzyme was able to oxidize Mn(2+) and still retain its ability to oxidize veratryl alcohol. Steady-state results indicate that the enzyme's ability to oxidize veratryl alcohol was lowered slightly. The K(m) for Mn(2+) was determined to be 1.57 mM and the k(cat) = 5.45 s(-1). These results indicate that the mutated lignin peroxidase is less effective in Mn(2+) oxidation that the wild type fungal enzyme. The pH optima of veratryl alcohol and Mn oxidation were altered by the mutation. They are one unit of pH value higher than those of recombinant H8 and wild type fungal Mn peroxidase isozyme H4. PMID- 11396963 TI - Preferential expression of reg I beta gene in human adult pancreas. AB - In human pancreas two genes, reg I alpha and reg I beta, have been characterized but only the reg I alpha protein has been isolated from human pancreatic secretion. To examine their respective physiological roles in fetal and adult pancreas we have compared the patterns of gene expression using a specific RT-PCR method. No progressive evolution in the two mRNAs levels was observed during fetal development (16--41 weeks). A discoordinate expression of the two genes was found with a higher level of reg I alpha mRNA in fetus and a higher level of regI beta in adult. In addition, if reg I alpha mRNA level was correlated with the expression of genes encoding exocrine proteins in adults, reg I beta mRNA level presented no correlation with any ductular, endocrine, or exocrine gene expression. In human pancreatic cell lines we showed the only expression of reg I beta gene and protein. All these data suggest that the two reg genes and proteins could play different roles in the pancreas. PMID- 11396964 TI - Induction of haptoglobin by all-trans retinoic acid in THP-1 human monocytic cell line. AB - Haptoglobin (Hp) is one of the acute-phase proteins and is mainly synthesized in the liver. During our study on the differentiation of leukemia cells, we have found that Hp is synthesized in human monocytic cells by all-trans retinoic acid (ATRA). The synthesis of Hp by ATRA is induced in a dose- and time-dependent manner. Hp cDNA cloned from ATRA-treated THP-1 cells corresponds to the Hp alpha 2(FS)-beta form. Whereas ATRA acted as a strong inducer in THP-1 cells, IL-1 beta, TNF-alpha, IL-6, and LPS had little effect on Hp gene expression in these cells. These findings suggest that THP-1 cells express the Hp gene through a signal pathway different from hepatocytes, and that ATRA is a potent Hp-inducer in these cells. PMID- 11396965 TI - Expression of the cell cycle phosphatase cdc25C is down-regulated by the tumor suppressor protein p53 but not by p73. AB - The cdc25C phosphatase dephosphorylates cdc2 kinase which then in complex with cyclin B can catalyse transition from the G(2) phase to mitosis. We demonstrate that transcription of cdc25C is repressed by p53 in a dose-dependent manner. In stably transfected DLD-1 colorectal adenocarcinoma cells, cdc25C expression is down-regulated when p53 is induced from a (tet)-off-regulated system. In contrast to p53, its homologue p73 is not able to down-modulate cdc25C expression. A previously identified site in the cdc25C promoter can bind p53 in vitro and, when placed in a heterologous construct, is able to activate transcription. However, transcriptional repression by p53 is not mediated through this site but is dependent on a segment containing three CCAAT-boxes. In general down-regulation of cdc25C transcription by reducing the levels of active cdc2 kinase contributes to G(2) arrest and G(2)/M checkpoint control. This reveals functional differences between p73 and p53 in regulating cell division. PMID- 11396966 TI - Differential N-demethylation of l-alpha-acetylmethadol (LAAM) and norLAAM by cytochrome P450s 2B6, 2C18, and 3A4. AB - Incubation of l-alpha-acetylmethadol (LAAM) or norLAAM with cDNA-expressed P450s 3A4, 2B6, and 2C18 produced significant N-demethylation products. P450s 2C19, 2C8, 3A5, 2C9, 3A7, 1A1, and 2D6 (norLAAM only), also produced detectable product. Coexpression of cytochrome b(5) enhanced LAAM N-demethylation, most dramatically for 3A4, but had marginal effects on norLAAM N-demethylation. Modeling total liver metabolism using immunoquantification and relative activity factors of P450s suggests contributions of P450 3A4 > 2B6 > 2C18, with the importance of 2B6 to 2C isozymes enhanced by relative activity factors. The ratio of dinorLAAM to norLAAM plus dinorLAAM formed from LAAM did not exceed 20%, and was isozyme and cytochrome b(5) coexpression dependent. This ratio decreased with concentration with 3A4, but was relatively constant for 2B6 and 2C18. The human liver microsomes substrate-concentration response was similar to cDNA-expressed 3A4, but the ratio was higher. Changes in the environment of cDNA-expressed 3A4 also effected the magnitude of the ratio, but not the concentration-dependent decrease. These studies show that the N-demethylation of LAAM and norLAAM is not restricted to P450 3A4, particularly P450s 2B6 and 2C18, and suggest that the mechanism of sequential metabolism for 3A4 differs from that of 2B6 and 2C18. PMID- 11396967 TI - A transgenic mouse expressing human CYP4B1 in the liver. AB - The human CYP4B1 protein was expressed in the liver of a transgenic mouse line under the control of the promoter of the human apolipoprotein E (apo E) gene. Hepatic microsomes of transgenic mice catalyzed omega-hydroxylation of lauric acid and also activated 2-aminofluorene (2-AF), which is a typical substrate for CYP4B1, to mutagenic compounds detected by an umu gene expression assay. These activities observed in transgenic mouse were efficiently inhibited by CYP4B1 antibody. However, such inhibition was not observed in control mice. This is the first report to indicate catalytic activities of human CYP4B1. For further characterization of human CYP4B1, a fusion protein of CYP4B1 and NADPH-P450 reductase was expressed in yeast cells. It was able to activate 2-AF and was also able to catalyze omega-hydroxylation of lauric acid. This transgenic mouse line and the recombinant fusion protein provide a useful tool to study human CYP4B1 and its relation to chemical toxicity and carcinogenesis. PMID- 11396968 TI - Transcriptional regulation of human 3'-phosphoadenosine 5'-phosphosulfate synthase 1. AB - Sulfonation, which is essential for normal growth, development and maintenance of the internal milieu, requires the universal sulfonate donor molecule 3' phosphoadenosine 5'-phosphosulfate (PAPS) produced from ATP and inorganic sulfate by two bifunctional PAPS synthase isozymes. The gene for PAPS synthase 1 containing neither a TATA nor a CCAAT box was found to be under the influence of the Sp1 family of transcription factors. Multiple GC/GT boxes are present in the proximal promoter region and deletion analysis implicated their involvement in transcription, a finding supported by mutational analysis of specific GC/GT boxes. Nuclear extract of SW13 cells, which highly express PAPS synthase 1, contains proteins that bind to probes possessing specific GC/GT boxes; furthermore, the presence of Sp1, Sp2, and Sp3 proteins in nuclear extracts was confirmed by supershift analysis. Cotransfection experiments using SL2 cells yielded additional support for the involvement of Sp1 in transcriptional regulation of the PAPS synthase 1 gene; the involvement of Sp2 and/or Sp3 is presently unclear. PMID- 11396969 TI - Influence of degradation on binding properties and biological activity of endomorphin 1. AB - The recently-isolated endogenous peptide endomorphin 1 has high affinity for the mu opioid receptor and plays an important role in analgesia. Several of its degradation products have been isolated from the central nervous system. Degradation products present structural similarities and may influence the receptor binding properties and biological activity of the parent compound. Therefore, we investigated how degradation of endomorphin 1 might influence ligand binding to the mu opioid receptor, the consequent activation of G proteins and its antinociceptive effect. Both N- and C-terminal truncation of endomorphin 1 resulted in peptides presenting considerably lower opioid receptor binding potency. None of these peptides had an effect on GTP binding, nor was able to produce analgesia, suggesting that degradation destroys the biological activity of endomorphin 1. PMID- 11396970 TI - Specific binding of baculoviruses displaying gp64 fusion proteins to mammalian cells. AB - Viral vectors displaying specific ligand binding moieties have raised an increasing interest in the area of targeted gene therapy. In this report, we describe baculovirus vectors displaying either a functional single chain antibody fragment (scFv) specific for the carcinoembryonic antigen (CEA) or the synthetic IgG binding domains (ZZ) derived from protein A of Staphylococcus aureus. In addition, the vectors were engineered to incorporate a reporter gene encoding the enhanced green fluorescent protein (EGFP) under the transcriptional regulation of the cytomegalovirus (CMV) IE promoter. Display of the targeting moieties on the viral surface was achieved through fusion to the N-terminus of gp64, the major envelope protein of the Autographa californica nuclear polyhedrosis virus (AcNPV). Specific binding of the gp64 fusion viruses to mammalian target cells was demonstrated by using monoclonal anti-gp64 antibodies followed by fluorescence and/or confocal microscopy. The anti-CEA scFv displaying baculovirus was shown to bind specifically to CEA expressing cells (PC-3). Similarly, the virus displaying the ZZ domains of protein A was targeted to BHK cells via binding of an appropriate IgG antibody. In all cases, the reporter gene was expressed in the transduced mammalian cells as shown by fluorescence microscopy and flow cytometric analyses. PMID- 11396971 TI - N- and C-terminal halves of human annexin VI differ in ability to form low pH induced ion channels. AB - Human recombinant annexin VI (AnxVI) or its N- (AnxVIA) and C-terminal (AnxVIB) fragments were expressed in E. coli. Their ability to form voltage-dependent ion channels in membranes, induced by low pH, was measured to verify the hypothesis that, upon acidification, the hydrophobicity of AnxVI at a specific domain significantly increases allowing the AnxVI interaction with lipids in a Ca(2+) independent manner. By theoretically analyzing changes in protein hydrophobicity, we found that hydrophobicity of AnxVIA significantly differed from that of AnxVIB at low pH. These predictions were confirmed experimentally by using planar lipid bilayers and liposome pull-down assay. We found striking difference between AnxVIA and AnxVIB in the ion channel activity, as well as in the membrane binding, suggesting that the halves of AnxVI maybe functionally different. Moreover, we calculated and predicted that the ion channel activity at low pH should appear in other human annexins, as AnxII, AnxV (as known), AnxVIII, and AnxXIII. The possibility that AnxVI acts as cytosolic component of a transmembrane pH-sensing mechanism is proposed. PMID- 11396972 TI - Genomic organization of the mouse plunc gene and expression in the developing airways and thymus. AB - Few genes have been isolated which display specific expression in the proximal airways. A recently identified mouse cDNA, plunc, appears to be confined to the upper airways and nasopharyngeal epithelium, and may prove a useful marker for these regions. We now report the genomic cloning and characterization of the mouse plunc gene as well as its developmental expression in the nasal and airway epithelium. We also report the novel finding that plunc is also expressed in the medullary compartment of the murine thymus. The mouse gene contains nine exons and the intron-exon boundaries are conserved with those in the human homologue. At e14.5 plunc is expressed in the nasal epithelium and several days later is seen in the thymic lobes, but not in the lining of the tracheobronchial tree. Expression in the trachea and main-stem bronchi first appears at 1--2 days after birth. Tracheobronchial expression persists at high levels throughout adulthood, as do regional areas of nasal and thymic expression. Finally, we show that the human homologue is expressed in bronchial epithelium, suggesting a transcript that is evolutionarily conserved in the mammalian airway. PMID- 11396973 TI - Expression of unphosphorylated form of human double-stranded RNA-activated protein kinase in Escherichia coli. AB - Interferon (IFN)-inducible, double-stranded (dsRNA)-activated protein kinase (PKR) is a key mediator of the antiviral and antiproliferative effects of IFN. PKR is present within cells in a latent state. In response to binding dsRNA, the enzyme becomes activated, causing autophosphorylation and an increase in specific kinase activity. In order to study PKR and its inhibitors, a large amount of the enzyme in its latent, unphosphorylated state is required. When PKR is fused to glutathione S-transferase (GST-PKR) and the fusion protein is expressed in Escherichia coli, the PKR obtained is fully activated by autophosphorylation. Therefore, we have developed an expression plasmid in which both GST-PKR and bacteriophage lambda protein phosphatase (lambda-PPase) genes were placed downstream of a T7 promoter. After induction of expression, unphosphorylated GST PKR was obtained in good yield, and purified to near homogeneity. The purified enzyme has dsRNA-dependent activation and phosphorylates the translation initiation factor eIF2 alpha. Using the recombinant protein, we analyzed the inhibition mechanisms of two viral inhibitors, vaccinia virus K3L protein and adenovirus virus-associated RNA I (VAI RNA). K3L inhibited both autophosphorylation of PKR and phosphorylation of eIF2 alpha, whereas VAI RNA inhibited only autophosphorylation. The separation of autophosphorylation and catalytic activity shows that the recombinant PKR is useful in analyzing the functions of PKR, its inhibitors, and its regulatory molecules. The coexpression system of protein kinase with lambda-PPase described here will be applicable to obtaining unphosphorylated and unactivated forms of other protein kinases. PMID- 11396974 TI - Antiproliferative effects of nitrosulindac on human colon adenocarcinoma cell lines. AB - Nonsteroidal anti-inflammatory drugs (NSAIDs) reduce the incidence of colon cancer, but their use is limited by toxicity in the gastrointestinal tract. The coupling of a nitric oxide-releasing moiety to NSAIDs strongly reduces these side effects. We demonstrated that the NO-releasing sulindac (nitrosulindac) has much more potent effects on colon adenocarcinoma cell lines compared to sulindac. Moreover, it could inhibit the growth of cells in soft agar experiments, demonstrating the antineoplastic activity at low concentration of nitrosulindac. However, this reduction in the growth of colon cancer cells seemed to be independent of the classical apoptosis pathway and could be explained by a cytostatic effect. Nitrosulindac caused a light perturbation of the cell cycle parameters not linked to a modification of the levels of p21 or the proliferating cell nuclear antigen. Moreover, neither sulindac, nor nitrosulindac, were able to inhibit the NF-kappa B pathway. These data suggested that nitrosulindac could be a better solution compared to other NSAIDs in the treatment of colon cancer. PMID- 11396975 TI - Possible implication of the Golgi apparatus casein kinase in the phosphorylation of vesicle docking protein p115 Ser-940: a study with peptide substrates. AB - Phosphorylation of human vescicle docking protein p115 at Ser-942 (homologous to Ser-940 in rat p115) promotes its dissociation from the Golgi membrane. Here we show that a peptide encompassing the 934--950 sequence of p115 is unaffected or poorly phosphorylated by a variety of Ser/Thr protein kinases with the notable exception of the Golgi apparatus casein kinase (G-CK) which phosphorylates it with an efficiency comparable to that of its optimal peptide substrates. In contrast phosphorylation of the p115 peptide by protein kinase CK2 is negligible compared to that of the specific peptide substrates of this kinase. Phosphorylation by G-CK is abolished if a conserved cluster of acidic residues at position between n + 4 and n + 9 (EDDDDE) is replaced by a neutral stretch (GAGAGA). These data strongly support the view that G-CK but not the other two classes of ubiquitous "casein kinases" (CK1 and CK2) is the natural phosphorylating agent of p115. PMID- 11396976 TI - Rabbit EPO gene and cDNA: expression of rabbit EPO after intramuscular injection of pDNA. AB - Erythropoietin (EPO) cDNA was cloned from kidney total RNA of a NZW rabbit. The cDNA comprises a 588-bp open reading frame encoding a 195 amino acid protein with distinguishable regions of high of homology to other mammalian EPOs. Intramuscular injection of mice with a rabbit EPO expression plasmid resulted in a significant hematocrit increase. A rabbit genomic DNA fragment was also cloned using the rabbit EPO cDNA. This 4312-bp genomic DNA fragment contains sequences homologous to the mouse EPO promoter and hypoxia-responsive enhancer. In addition, the genomic DNA also presents a high degree of conservation to other regions involved in hypoxia response. Sequence divergence in the 3' UTR may indicate differences in regulation of mRNA stability or response to low oxygen tension. PMID- 11396977 TI - Recombinant expression and purification of human androgen receptor in a baculovirus system. AB - A full-length human androgen receptor (hAR) cDNA was used to produce recombinant baculovirus. Spodoptera frugiperda (Sf9) cells infected with this virus expressed protein with an N-terminal hexahistidine tag (His(6)-hAR) in soluble and insoluble forms. The soluble cytosolic His(6)-hAR demonstrated similar association and dissociation half-times for mibolerone, similar binding affinity for mibolerone, and similar steroid specificity as bona fide AR. Under native conditions, the soluble cytosolic His(6)-hAR was purified to apparent homogeneity in the presence of dihydrotestosterone, using metal ion affinity chromatography. The insoluble pellet fraction was solubilized with strong denaturant 6 M guanidine HCl, and His(6)-hAR was purified from it in the presence of 6 M guanidine HCl. Both the solubilized crude pellet fraction and the solubilized/purified His(6)-hAR could be renatured to bind mibolerone. The baculovirus system will therefore provide an efficient means for producing hAR for ligand-binding assays, as well as purifying hAR for detailed molecular analyses. PMID- 11396978 TI - Identification of a novel phosphorylation site in human androgen receptor by mass spectrometry. AB - An N-terminal hexahistidine-tagged full-length human androgen receptor protein (His(6)-hAR) was overexpressed and purified to apparent homogeneity in the presence of dihydrotestosterone (DHT) in our previous studies. In-gel trypsin digestion of the purified DHT-bound His(6)-hAR, and tryptic peptide mapping using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI/TOF-MS), detected a total of 17 peptides (21% coverage of hAR) with 9 peptides originating from the ligand-binding domain (LBD, 31% coverage of LBD). Amino acid sequencing analysis of the tryptic peptides from a separate in-gel digestion of the His(6)-hAR, using HPLC-coupled electrospray ionization ion trap mass spectrometry (LC/ESI-ITMS and MS/MS), unambiguously confirmed 21 peptides with 19% coverage of the hAR, of which 11 peptides originated from the LBD (35% coverage of LBD). These 21 peptides included 11 out of the 17 peptides detected by MALDI/TOF-MS. In addition, a novel serine phosphorylation site (Ser(308)) within the N-terminal transactivation domain of hAR was identified. PMID- 11396979 TI - Plant sterol 14 alpha-demethylase affinity for azole fungicides. AB - Azole fungicides were thought to have much greater affinity for the fungal cytochrome P450 enzyme, sterol 14 alpha-demthylase (CYP51) than the plant orthologue. Using purified CYP51 from the plant Sorghum bicolor L Moenech, a direct comparison of the sensitivity to the fungicides triadimenol and tebuconazole has been carried out. S. bicolor CYP51 was purified to homogenity as determined by SDS--PAGE and specific heme content. Addition of the azole fungicides triadimenol and tebuconazole induced type II spectral changes, with saturation occurring at equimolar azole/P450 concentrations. Inhibition of reconstituted activities revealed only a threefold insensitivity of the plant CYP51 compared to a fungal CYP51, from the phytopathogen Ustilago maydis, as judged by IC(50) values. The implications for fungicide mode of action and application are discussed. PMID- 11396980 TI - Altered expression of the fragile histidine triad gene in primary gastric adenocarcinomas. AB - Genomic alterations and abnormal expression of the fragile histidine triad (FHIT) gene in gastric carcinomas were examined to determine whether the FHIT gene is actually a frequent target for alteration during gastric carcinogenesis. To correlate DNA and RNA lesions of the FHIT gene with the effect on FHIT protein expression, we have investigated the FHIT gene for loss of heterozygosity (LOH), aberrant transcripts, point mutations, and protein expression in 35 gastric adenocarcinomas. Allelic loss at D3S1300 was detected in 7 of 33 (21%) informative cases. Aberrant transcripts, with deletions and/or insertions, were observed in 20 of 35 (57.1%) cases and resulted from alternative splicing through exon skipping and/or insertion of the FHIT intron 5 sequence or activation of the cryptic splice site. Point mutations were not found in the FHIT coding region but detected in noncoding exon 2, 3, 4, or 5 of eight aberrant transcripts. Significant reduction of FHIT protein expression was observed in 22 of 35 (62.9%) cases. Aberrant FHIT transcription was shown to be associated with loss of FHIT protein expression. However, aberrant FHIT transcripts themselves were not associated with any clinicopathological parameters, such as age, sex, tumor site, or clinical stage. Moreover, there was no association between the presence of LOH at D3S1300 and the expression of aberrant FHIT transcripts. Nevertheless, high frequency of aberrant FHIT transcripts, significant rate of LOH at D3S1300, and altered expression of the FHIT protein indicate that alterations of the FHIT gene can play an important role in gastric carcinogenesis. PMID- 11396981 TI - An analog of the human albumin N-terminus (Asp-Ala-His-Lys) prevents formation of copper-induced reactive oxygen species. AB - Copper mobilization and redox activity form damaging reactive oxygen species (ROS) and are implicated in the pathogenesis of ischemia-reperfusion injury, chronic inflammation, Alzheimer's disease, aging, and cancer. Protein sequestration of Cu(II) ions has been shown to prevent ROS-generating reactions. The first four amino acids of the N-terminus of human albumin, Asp-Ala-His-Lys (DAHK), form a tight binding site for Cu(II) ions. We synthesized several analogs, including the enantiomer d-DAHK, to study their effects on copper induced hydroxyl radical and superoxide formation in the presence of ascorbate. d DAHK prevented thiobarbituric acid-reactive species (TBARS) formation within physiological and acidic pH ranges (7.5-6.5) and inhibited low-density lipoprotein lipid peroxidation. A d-DAHK/Cu complex exhibited superoxide dismutase-like activity by significantly inhibiting superoxide formation. These in vitro results suggest that d-DAHK may shift the Cu(II)-binding equilibrium from the exchangeable Cu(II) pool to the tightly-bound, nonexchangeable pool, prevent ROS formation, and potentially provide therapeutic benefit for ROS related diseases. PMID- 11396982 TI - Total calcium ultrastructure: advances in excitable cells. AB - This is a review which is written on the basis of a cell calcium lecture delivered on 22 July 2000 at the European Research Meeting 'Calcium as a molecule of cellular integration'. PMID- 11396983 TI - Functional expression of recombinant type 1 ryanodine receptor in insect cells. AB - We have investigated the biochemical properties of the rabbit ryanodine receptor type 1 (RyR1) from skeletal muscle functionally expressed in insect sf 21 cells infected with recombinant baculovirus. Equilibrium [3H]ryanodine binding assays applied to total membrane fractions from sf 21 cells expressing recombinant RyR1 showed a non-hyperbolic saturation curve (Hill coefficient = 2.1). The [3H]ryanodine binding was enhanced by 1 mM AMP-PCP and 10 mM caffeine, whereas 10 mM Mg(2+) and 5 microM ruthenium red reduced the specific binding. The dependence of [3H]ryanodine binding on ionic strength showed positive cooperativity (Hill coefficient = 2.2) with a plateau at 1 M KCl. The recombinant RyR1 showed a bell shaped [3H]ryanodine binding curve when free [Ca(2+)] was increased, with an optimal concentration around 100 microM.Confocal microscopy studies using the Ca(2+) ATPase selective inhibitor, thapsigargin coupled to fluorescein and ryanodine coupled to Texas red demonstrated that the recombinant RyR1 and the Ca(2+) ATPase co-localize to the same intracellular membrane. No significant RyR1 fluorescence was observed at the plasma membrane.Fluo-4-loaded sf 21 cells expressing recombinant RyR1 responded to activating-low ryanodine concentrations (100 nM) or caffeine (10 mM) with a sharp rise in intracellular Ca2 followed by a sustained phase, in contrast, sf 21 cells expressing the human bradykinin type 2 receptor did not respond to ryanodine or caffeine.These results demonstrate the expression of recombinant RyR1 in sf 21 cells with functional properties similar to what has been previously reported for native RyR1 in mammalian tissues, however, some differences were observed in [3H]ryanodine binding assays compared to native rabbit RyR1. Hence, the baculovirus expression system provides a generous source of protein to accomplish structure-function studies and an excellent model to assess functional properties of wild type and mutant RyR1. PMID- 11396984 TI - Simulation of cardiac work transitions, in vitro: effects of simultaneous Ca2+ and ATPase additions on isolated porcine heart mitochondria. AB - During increases in cardiac work there are net increases in cytosolic [Ca(2+)] and ATP hydrolysis by myofiliments and ion transport ATPases. However, it is still unclear what role Ca(2+)or the ATP hydrolysis products, ADP and Pi, have on the regulation of mitochondrial ATP production. In this study, work jumps were simulated by simultaneous additions of Ca(2+) and ATPase to porcine heart mitochondria. The net effects on the mitochondrial ATP production were monitored by simultaneously monitoring respiration (mVo2), [NADH], [ADP] and membrane potential (deltapsi) at 37 degrees C. Addition of exogenous ATPase (300 mlU.ml( 1))]ATP (3.4 mM) was used to generate a 'resting' background production of ADP. This resting metabolic rate was 200% higher than the quiescent rate while [NADH] and deltapsi were reduced. Subsequent ATPase additions (1.3IU.ml(-)) were made with varying amounts of Ca(2+)(0 to 535 nM) to simulate step increases in cardiac work. Ca(2+) additions increased mVo2 and depolarized deltapsi, and were consistent with an activation of Fo/F1)ATPase. In contrast, Ca(2+) reduced the [NADH] response to the ATPase addition, consistent with Ca(2+)-sensitive dehydrogenase activity (CaDH). The calculated free ADP response to ATPase decreased ?2-fold in the presence of Ca(2+). The addition of 172nM free Ca(2+)] ATPase increased mVo2 by 300% (P<0.05, n=8) while deltapsi decreased by 14.9+/ 0.1 mV without changes in [NADH] (P > or =0.05, n=8), consistent with working heart preparations. The addition of Ca(2+) and ATPase combined increased the mitochondrial ATP production rate with changes in deltapsi, NADH and [ADP], consistent with an activation of CaDH and F o /F(1)ATPase activity. These balancing effects of ATPase activity and [Ca(2+)] may explain several aspects of metabolic regulation in the heart during work transitions in vivo. PMID- 11396985 TI - Endothelial intracellular Ca2+ release following monocyte adhesion is required for the transendothelial migration of monocytes. AB - Although molecular changes accompanying leukocyte extravasation have been investigated intensively, the particular events following leukocyte adhesion and leading to the actual transendothelial migration process remain largely unknown. To characterize intraendothelial signals elicited by leukocyte adhesion and functionally required for their transmigration, we recorded endothelial free cytosolic intracellular Ca(2+)levels ([Ca(2+)]i) during the course of leukocyte adhesion. We show that monocyte and granulocyte adhesion induced Ca(2+)transients in either untreated or TNF-alpha-stimulated microvascular endothelial cells (HMEC 1). The functional significance of these [Ca(2+)]i rises was demonstrated by treating filter-grown endothelial monolayers with BAPTA/AM. This in traendothelial Ca(2+)chelation left monocyte adhesion basically unaffected, but caused a significant and dose-dependent reduction of the transendothelial migration of monocytes. Granulocyte diapedesis, on the other hand, was hardly modified. Thapsigargin-treatment of endothelial cells almost completely inhibited the transmigration of monocytes suggesting that the necessary Ca(2+)transients depended on a release from intracellular Ca(2+)stores. Our results thus show that the transmigration of monocytes through endothelial monolayers of microvascular origin is favoured by an increase of the intraendothelial [Ca(2+)]i induced by leukocyte adhesion to the endothelial cells. PMID- 11396986 TI - The effect of calcium pump inhibitors on the response of intracellular calcium to caffeine in snail neurones. AB - We have measured intracellular free calcium ([Ca(2+)]i) using Fura-2 or Ca(2+) sensitive microelectrodes in voltage-clamped neurones of the snail, Helix aspersa. Caffeine-induced transient increases in [Ca(2+)]i were normally followed by a brief fall of [Ca(2+)]i below its pre-caffeine level. We investigated the cause of this undershoot by raising [Ca(2+)]i; and by inhibiting the plasma membrane or endoplasmic reticulum Ca ATPases (PMCA or SERCA respectively). When the cell membrane potential was decreased from -60 to -25mV, steady-state [Ca(2+)]i increased. The caffeine-induced transients were smaller while the undershoots were larger than in control conditions. When the PMCA was inhibited by high pH the steady-state [Ca(2+)]i increased by 100-400nM. The caffeine induced [Ca(2+)]i increase and the subsequent undershoot both became larger. Injection of orthovanadate, which inhibits the PMCA and increases [Ca(2+)]i, did not block either effect of caffeine. But when the SERCA was inhibited by cyclopiazonic acid the undershoot disappeared. The phosphodiesterase inhibitor IBMX did not influence the undershoot. These results suggest that the undershoot is generated by the Ca(2+)] ATPase of the stores rather than that of the plasma membrane. Since the undershoot increased as [Ca(2+)]i increased, we conclude that at higher levels of [Ca(2+)]i the stores refill more rapidly. PMID- 11396987 TI - Delayed activation of the plasma membrane calcium pump by a sudden increase in Ca2+: fast pumps reside in fast cells. AB - There are four genes encoding isoforms of the plasma membrane Ca(2+) pump (PMCA). PMCA variability is increased by the presence of two splicing sites. Functional differences between the variants of PMCA have been described, but little is known about the adaptive advantages of this great diversity of pumps. In this paper we studied how the different isoforms respond to a sudden increase in Ca(2+) concentration. We found that different PMCAs are activated by Ca(2+) at different rates, PMCA 3f and 2a being the fastest, and 4b the slowest. The rate of activation by Ca(2+) depends both on the rate of calmodulin binding and the magnitude of the activation by calmodulin. We found that 2a is located in heart and the stereocilia of inner ear hair cells, 3f in skeletal muscle and 4b was identified in Jurkat cells. Both cardiac and skeletal muscle, and stereocilia recover very rapidly after a cytoplasmic Ca(2+)peak, while in Jurkat cells the recovery takes up to a minute. In stereocilia, 2a is the only method for export of Ca(2+), making the analysis of them unusually straightforward. This indicates that these rates of PMCA activation by Ca(2+) are correlated with the speed of Ca(2+) concentration decay after a Ca2 spike in the cells in which these variants of PMCA are expressed. The results suggest that the type of PMCA expressed will correspond with the speed of Ca(2+) signals in the cell. PMID- 11396988 TI - Restoration of calcium handling properties of adult cardiac myocytes from hypertrophied hearts. AB - Reductions in cardiac sarcoplasmic reticulum calcium-ATPase (Serca2a) levels are thought to underlie the prolonged calcium (Ca(2+)) transients and consequent reduced contractile performance seen in human cardiac hypertrophy and heart failure. In freshly isolated cardiac myocytes from rats with monocrotaline induced right ventricular hypertrophy we found reduced sarcoplasmic reticulum Serca2a expression and prolonged Ca(2+)transients, characteristic of hypertrophic cardiac disease. Modulation of intracellular Ca(2+)levels, Ca(2+) kinetics or Ca(2+)sensitivity is the focus of many current therapeutic approaches to improve contractile performance in the hypertrophic or failing heart. However, the functional effects of increasing Serca2a expression on Ca(2+) handling properties in myocytes from an animal model of cardiac hypertrophy are largely unknown. Here, we describe enhancement of the deficient Ca(2+) handling properties evident in myocytes from hypertrophied hearts following adenoviral-mediated transfer of the human Serca2a gene to these myocytes. These results highlight the importance of Serca2a deficiencies in the hypertrophic phenotype of cardiac muscle and suggest a simple, effective approach for manipulation of normal cardiac function. PMID- 11396989 TI - Potentiation by stannous chloride of calcium entry into osteoblastic MC3T3-E1 cells through voltage-dependent L-type calcium channels. AB - The present study was undertaken to confirm that L-type Ca(2+) channels are involved in Ca(2+) entry into osteoblastic MC3T3-E1 cells and to examine the effect of SnCl2, a Ca(2+)]-channel activator, on the intracellular Ca(2+)concentration ([Ca(2+)]i). High K(+)concentration-dependently raised the [Ca(2+)]i. All of the L-type Ca(2+)channel blockers used here, such as nifedipine, nicardipine, verapamil, and diltiazem, and CdCl2 (a non-selective blocker) inhibited the high K(+)-induced [Ca(2+)]i rise, but v-conotoxin GVIA (an N-type blocker) and NiCl2(a T-type blocker) had no effect. Application of SnCl2 alone did not change the [Ca(2+)]i. However, in the presence of high K(+), SnCl2 enhanced the high K(+)-induced [Ca(2+)]i rise, which was inhibited by Ca(2+)] free medium or nifedipine. In the case where high K(+)was applied prior to SnCl2, SnCl2 alone raised the [Ca(2+)]i by itself. In conclusion, MC3T3-E1 cells possess the voltage-dependent L-type Ca(2+)] channels and SnCl2 facilitates the Ca(2+) entry through the L-type ones under the condition of the membrane depolarization. There is the possibility that Ca(2+) release from intracellular Ca(2+) stores is involved in the action of SnCl2. PMID- 11396990 TI - Upregulation of bone morphogenetic protein GDF-3/Vgr-2 expression in adipose tissue of FABP4/aP2 null mice. AB - High-fat-fed C57Bl/6J FABP4/aP2 null mice develop obesity but not the related hyperglycemia or hyperinsulinemia characteristic of type II diabetes. FABP4/aP2 protein's function to bind fatty acids in the adipocytes may promote total body energy homeostasis by linking energy depots to the ability to express signaling molecules similar to leptin. To test this hypothesis, proteomic analysis of serum proteins from high-fat-fed wild-type and FABP4/aP2 null mice revealed that the GDF-3/Vgr-2 protein, a bone morphogenetic protein, was upregulated in C57Bl/6J FABP4/aP2 null mice. The increase in serum GDF-3/Vgr-2 protein was correlated with a 27-fold increase in adipose GDF-3/Vgr-2 mRNA. In contrast, leptin expression was unaltered between FABP4/aP2 null and wild-type animals. The expression of GDF-3/Vgr-2 mRNA was not substantially different in adipose tissue of db/db and tb/tb mice compared to wild-type controls. The expression of GDF 3/Vgr-2 mRNA was dependent upon the age and diet of the animals, declining as a function of age in high-fat-fed wild-type animals while increasing in the FABP4/aP2 null strain. These results identify GDF-3/Vgr-2 as an age- and fat regulated, adipose-derived cytokine suggesting a linkage between adipocyte fatty acid metabolism and the expression of the bone morphogenetic family of differentiation regulators. PMID- 11396991 TI - Expression and release of stable and active forms of murine granulocyte macrophage colony-stimulating factor (mGM-CSF) targeted to different subcellular compartments. AB - Cytokines have been used for several years as immunomodulators. However, one of the main drawbacks of systemically applied cytokines is their high toxicity. In addition, cytokines work in a paracrine form and frequently after cell-to-cell interaction. Therefore, a very restricted release of cytokines-in time and space could be desired for most of their therapeutic applications. The murine granulocyte-macrophage colony-stimulating factor (mGM-CSF) is one of the most promising cytokine candidates for cancer immunotherapy and as an adjuvant of DNA vaccines. With the aim of improving the administration and release of cytokines in a very restricted area, we have designed vectors expressing the mGM-CSF cDNA with different localization signals. Using this strategy we have shown that cytokines can be expressed and targeted to different subcellular compartments (i.e. the cytoplasm, the endoplasmic reticulum and the nucleus), stored inside the cells and released after cell lysis as stable active proteins. Moreover, a plasma membrane targeted form of mGM-CSF displayed substantial amount of biological activity. These vectors could have potential applications in immunotherapy for tumours and DNA vaccination protocols. PMID- 11396992 TI - Expression of interleukin 7 (IL-7) mRNA and protein in the normal adult human liver: implications for extrathymic T cell development. AB - Interleukin-7 (IL-7) has been shown to play an essential role in T-cell development. Recombinase-activating gene (RAG)-1, RAG-2 and pre-TCR-alpha expression in the normal adult human liver (AHL), together with the presence of lymphoid-haematopoietic progenitors, is strong evidence that the AHL supports T cell maturation. We investigated IL-7 mRNA and protein levels in order to determine whether AHL could support T lymphocyte differentiation. Biopsies were snap frozen, powdered, and RNA/protein extracted. Reverse transcriptase polymerase chain reaction was used to detect IL-7 using primers that amplified 620 base pair (bp) fragments and other smaller transcripts. A sandwich enzyme linked immunosorbent assay was developed to quantify IL-7 protein in homogenates. The anatomic distribution of IL-7-secreting cells was determined by immunohistochemistry. IL-7-specific product (620 bp) was detected in nine of ten samples, with six also positive for a smaller splice-variant (488 bp). Levels of the 620 bp product were 2.5 times greater than the 488 bp fragment. IL-7 protein was detected in all samples (range 18.47-76.93 pg/100 mg tissue). Immunohistochemistry demonstrated IL-7 protein in discrete cells of lymphoid morphology, widely distributed throughout the parenchyma and within portal tracts. Large populations of innate T cells are found in normal AHL, some of which may differentiate locally. The presence of IL-7 RNA and protein throughout normal hepatic tissue provides evidence that the normal AHL is a suitable microenvironment for T cell differentiation. PMID- 11396993 TI - Expression of IL-1 beta and IL-8 by human gingival epithelial cells in response to Actinobacillus actinomycetemcomitans. AB - The interaction between epithelial cells and microorganisms is the most important step in bacterial infections. Epithelial cells in response to exposure to pathogenic bacteria produce cytokines that initiate inflammation. However, little is known about the cytokine response of gingival epithelial cells to periodontopathogenic bacteria. Actinobacillus actinomycetemcomitans is thought to play a significant role in the initiation of periodontitis because of its bacteriological characteristics. In the present study, we investigated the cytokine induction by human gingival epithelial cells (HGEC) following exposure to A. actinomycetemcomitans in comparison with human gingival fibroblasts (HGF) in culture. Northern blot analysis showed that mRNAs of interleukin 1beta (IL 1beta) and IL-8, but not IL-6, in HGEC were induced in response to A. actinomycetemcomitans. Secretion of IL-8 by HGEC was also increased following A. actinomycetemcomitans challenge, whereas production of IL-1beta could not be detected. The levels of IL-8 and its mRNA were increased depending on the concentration of A. actinomycetemcomitans. The co-culture with HGF and A. actinomycetemcomitans resulted in an increase in the levels of IL-6 and IL-8 mRNA in HGF. However, HGF exposed to A. actinomycetemcomitans, showed no expression of IL-1beta mRNA. These findings demonstrated that HGEC and HGF stimulated with A. actinomycetemcomitans have different profiles in cytokine mRNA expression. Furthermore, A. actinomycetemcomitans may play an important role in amplifying the local immune response and in initiating inflammatory reaction through release of IL-8 from gingival epithelial cells. PMID- 11396994 TI - Gender differences in the inflammatory response and survival following haemorrhage and subsequent sepsis. AB - Studies have shown gender dimorphism in cell-mediated immune responses following haemorrhage, with depressed responses in young males and maintained or enhanced responses in proestrus females. However, it remains unknown whether or not the sexually dimorphic immune response to haemorrhage provides any protection against a subsequent in vivo polymicrobial septic challenge. To study this, male and proestrus female C3H/HeN mice were subjected to haemorrhage (35+/-5 mmHg for 90 min followed by fluid resuscitation) or sham operation. Twenty-four hours thereafter, all mice were subjected to polymicrobial sepsis by cecal ligation and puncture (CLP) and survival was assessed over a 10 day period. Haemorrhage prior to CLP significantly increased mortality in males as compared to shams. In contrast, mortality in females following CLP was comparable between the sham and haemorrhage groups. Plasma levels of interleukin (IL-)6, tumour necrosis factor (TNF)-alpha and prostaglandin E(2)(PGE(2)) at 5 h after CLP were significantly increased in males subjected to prior haemorrhage. In contrast, plasma levels of IL-6 and TNF-alpha in females did not increase under such conditions. PGE(2)levels were comparable in males and females following CLP, however prior haemorrhage significantly reduced PGE(2)levels in females, whereas no change was observed in males. Liver and splenic expression of cyclooxygenase-2 protein paralleled the changes in plasma PGE(2). Female sex hormones, therefore, appear to play an important role not only in maintaining immune function following haemorrhage, but also provide a survival advantage against subsequent septic challenge. PMID- 11396995 TI - Concentrations of hepatocyte growth factor in cerebrospinal fluid under normal and different pathological conditions. AB - Hepatocyte growth factor (HGF) and its specific receptor, MET, are expressed in the developing and adult mammalian brain. Recent studies have shown a neurotrophic activity of HGF in the nervous system. The present study focused on HGF concentrations in the cerebrospinal fluid (CSF) and serum in normal persons and in different central nervous system (CNS) diseases considering blood-CSF barrier (BCB) function. Concentrations of HGF were analyzed using an enzyme linked immunosorbent assay (ELISA). HGF was present in normal human CSF (346+/ 126 pg/ml) representing approximately half of the HGF serum concentrations. The CSF HGF levels were not significantly changed in chronic CNS disease and in aseptic meningitis (419+/-71 pg/ml), but significantly increased in patients with bacterial meningitis (6101+/- 5200 pg/ml). The HGF levels in CSF were not influenced by increased serum concentrations in patients with normal or mildly affected BCB function. The results show that HGF is present in normal CSF and does not appear to cross the CSF barrier significantly unless it is severely disrupted. So far, strong increases of HGF concentration in CSF are only present in acute bacterial meningitis. PMID- 11396996 TI - Molecular cloning of gerbil interleukin 12 and its expression as a bioactive single-chain protein. AB - Complementary DNAs coding for gerbil interleukin 12 (IL-12) p40/p35 subunits were cloned by a combination of cross species reverse transcriptase-polymerase chain reaction (RT-PCR) and 3' rapid amplification of cDNA ends (RACE) techniques. IL 12 p40/p35 had 79% nucleotide identity and 81% amino acid homology to mouse IL-12 p40/p35. The p40/p35 subunits were expressed as a single polypeptide separated by a short hinge sequence that allowed for proper folding and assembly. COS-7 cells transfected with DNA encoding the single-chain gerbil IL-12 (pSCjIL12) secreted high levels of the protein which stimulated proliferation of ConA-activated gerbil spleen lymphoblasts in a dose-dependent manner. PMID- 11396997 TI - Granulocyte-macrophage colony-stimulating factor (GM-CSF) decreases left ventricular function. An echocardiographic study in cancer patients. AB - To study the effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) on the heart, echocardiographic assessments of left ventricular (LV) end diastolic and end-systolic (ES) diameters (D), ejection fraction (EF) and cardiac output (CO) were done in six male patients (28-70 years of age) with advanced sarcoma (Group 1), prior to (day -1-0), during (day 7-9) and after (day 20-21) a first course of i.v. doxorubicin (day 0) without GM-CSF and a second course (3 weeks after the first one) with GM-CSF 250 microg/m(2)subcutaneously and daily from day 1-11. A similar study was done in ten female patients with advanced breast cancer (31-58 years of age, Group 2) for a first course of doxorubicin+cyclophosphamide with GM-CSF (same schedule as in Group 1). As compared to the mean of values prior to and after the course with GM-CSF in Group 1 and 2, the LVESD during GM-CSF administration transiently increased by median 6% (range -19 to 30%, P<0.05) vs -9% (-21 to 6%, not significant) in the first course without GM-CSF in Group 1 (P<0.05 between courses). The CO and EF tended to decrease during GM-CSF. GM-CSF thus causes a small and transient decrease of LV contractility. PMID- 11396998 TI - Maximal local and minimal systemic cytokine response to colorectal surgery: the influence of perioperative filgrastim. AB - Thirty consecutive patients scheduled for elective colorectal surgery were prospectively randomized to receive either filgrastim [the recombinant human form of granulocyte colony-stimulating factor (r-metHu-G-CSF)] or placebo blindly. The levels of interleukin (IL-)1beta, tumour necrosis factor-alpha (TNF-alpha), IL-6, IL-8, transforming growth factor-beta (TGF-beta), and IL-10 were determined 5 and 24 h postoperatively from peripheral blood, peritoneal fluid, and wound fluid. The concentrations of all the measured cytokines were enormously higher locally at the operative site than in the systemic circulation. The only difference between the two medication groups was the lower concentration of IL-8 in peripheral blood in the filgrastim-treated patients. The present study shows abundant release of pro- and anti-inflammatory cytokines into the wound and the peritoneal cavity after abdominal surgery. The systemic response to surgery seems to be a secondary and minor reflection of local events. Filgrastim did not have any effect on the studied local cytokine levels at the operative site. PMID- 11396999 TI - Dosage-dependent gene regulation in multicellular eukaryotes: implications for dosage compensation, aneuploid syndromes, and quantitative traits. AB - Evidence from a variety of data suggests that regulatory mechanisms in multicellular eukaryotes have evolved in such a manner that the stoichiometric relationship of the components of regulatory complexes affects target gene expression. This type of mechanism sets the level of gene expression and, as a consequence, the phenotypic characteristics. Because many types of regulatory processes exhibit dosage-dependent behavior, they would impact quantitative traits and contribute to their multigenic control in a semidominant fashion. Many dosage-dependent effects would also account for the extensive modulation of gene expression throughout the genome that occurs when chromosomes are added to or subtracted from the karyotype (aneuploidy). Moreover, because the majority of dosage-dependent regulators act negatively, this property can account for the up regulation of genes in monosomics and hemizygous sex chromosomes to achieve dosage compensation. PMID- 11397000 TI - Differential expression of Eph receptors and ephrins correlates with the formation of topographic projections in primary and secondary visual circuits of the embryonic chick forebrain. AB - Repulsion plays a fundamental role in the establishment of a topographic map of the chick retinotectal projections. This has been highlighted by studies demonstrating the role of opposing gradients of the EphA3 receptor tyrosine kinase on retinal axons and two of its ligands, ephrin-A2 and ephrin-A5, in the tectum. We have analyzed the distribution of these two ephrins in other retinorecipient structures in the chick diencephalon and mesencephalon during the period when visual connections are being established. We have found that both ephrin-A2 and ephrin-A5 and their receptors EphA4 and EphA7 are expressed in gradients whose orientation is consistent with the topography of the nasotemporal axis of the respective retinofugal projections. In addition, their distribution suggests that receptor-ligand interactions may be involved in the organization of connections between the different primary visual centers and, thus, in the topographic organization of secondary visual projections. Interestingly, where projections lack a clear topographic representation, a uniform expression of the Eph-ephrin molecules was observed. Finally, we also show that a similar patterning mechanism may be implicated in the transfer of visual information to the telencephalon. These results suggest a conserved function for EphA receptors and their ligands in the elaboration of topographic maps at multiple levels of the visual pathway. PMID- 11397001 TI - Distinct enhancer elements control Hex expression during gastrulation and early organogenesis. AB - In the mouse, embryological and genetic studies have indicated that two spatially distinct signalling centres, the anterior visceral endoderm and the node and its derivatives, are required for the correct patterning of the anterior neural ectoderm. The divergent homeobox gene Hex is expressed in the anterior visceral endoderm, in the node (transiently), and in the anterior definitive endoderm. Other sites of Hex expression include the liver and thyroid primordia and the endothelial cell precursors. We have used transgenic analysis to map the cis acting regulatory elements controlling Hex expression during early mouse development. A 4.2-kb upstream region is important for Hex expression in the endothelial cell precursors, liver, and thyroid, and a 633-bp intronic fragment is both necessary and sufficient for Hex expression in the anterior visceral endoderm and the anterior definitive endoderm. These same regions drive expression in homologous structures in Xenopus laevis, indicating conservation of these regulatory regions in vertebrates. Analysis of the anterior visceral endoderm/anterior definitive endoderm enhancer identifies a repressor region that is required to downregulate Hex expression in the node once the anterior definitive endoderm has formed. This analysis also reveals that the initiation of Hex expression in the anterior visceral endoderm and axial mesendoderm requires common elements, but maintenance of expression is regulated independently in these tissues. PMID- 11397002 TI - Inner cell mass-specific expression of a cell adhesion molecule (PECAM-1/CD31) in the mouse blastocyst. AB - Platelet/Endothelial Cell Adhesion Molecule-1 (PECAM-1 or CD31) is thought to be a vascular-specific protein, but its function has not been clearly defined. Here, we demonstrate by using confocal immunofluorescence microscopy that PECAM-1 is first detected in the mouse blastocyst, which contains no vascular cells, and its expression is restricted to the pluripotent inner cell mass (ICM) cells. Expression is localized to cell-cell borders of the ICM and is detected at the very first signs of blastocoel formation. Consistent with these observations is that embryonic transcripts of PECAM-1 mRNA, as detected by RT-PCR, greatly increase during the morula-to-blastocyst transition and seven of the eight known alternatively spliced isoforms of PECAM-1 are expressed in the blastocyst. The synthesis of PECAM-1 is independent of compaction, cytokinesis, and DNA replication, as it is detected in embryos that are chronologically at the blastocyst stage following culture of 8-cell embryos in Ca2+-free medium, or medium containing cytochalasin D or aphidicolin, respectively. By the late blastocyst stage, PECAM-1 expression is restricted to the pluripotent epiblast, at which point it has a mutually exclusive expression pattern to that of type IV collagen, a basement membrane marker. The reduction in PECAM-1 transcripts in retinoic acid-induced differentiation of F9 teratocarcinoma cells, a model of epiblast-to-primitive endoderm differentiation, confirmed the epiblast-specific expression of PECAM-1. By the egg cylinder stage of development, at which point the epiblast is no longer pluripotent, PECAM-1 is not detected. This ICM-specific pattern of expression suggests a novel developmental role of PECAM-1 that is independent of its function in vascular ontogeny. PMID- 11397003 TI - Bmp2b and Oep promote early myocardial differentiation through their regulation of gata5. AB - Members of both the bone morphogenetic protein (Bmp) and EGF-CFC families have been implicated in vertebrate myocardial development. Zebrafish swirl (swr) encodes Bmp2b, a member of the Bmp family required for patterning the dorsoventral axis. Zebrafish one-eyed pinhead (oep) encodes a maternally and zygotically expressed member of the EGF-CFC family essential for Nodal signaling. Both swr/bmp2b and oep mutants exhibit severe defects in myocardial development. swr/bmp2b mutants exhibit reduced or absent expression of nkx2.5, an early marker of the myocardial precursors. Embryos lacking zygotic oep (Zoep mutants) display cardia bifida and, as we show here, also display reduced or absent nkx2.5 expression. Recently, we have demonstrated that the zinc finger transcription factor Gata5 is an essential regulator of nkx2.5 expression. In this paper, we investigate the relationships between bmp2b, oep, gata5, and nkx2.5. We show that both swr/bmp2b and Zoep mutants exhibit defects in gata5 expression in the myocardial precursors. Forced expression of gata5 in swr/bmp2b and Zoep mutants restores robust nkx2.5 expression. Moreover, overexpression of gata5 in Zoep mutants restores expression of cmlc1, a myocardial sarcomeric gene. These results indicate that both Bmp2b and Oep regulate gata5 expression in the myocardial precursors, and that Gata5 does not require Bmp2b or Oep to promote early myocardial differentiation. We conclude that Bmp2b and Oep function at least partly through Gata5 to regulate nkx2.5 expression and promote myocardial differentiation. We integrate these and other data to propose a pathway of the molecular events regulating early myocardial differentiation in zebrafish. PMID- 11397004 TI - Mouse ovarian germ cell cysts undergo programmed breakdown to form primordial follicles. AB - In many organisms, early germline development takes place within cysts of interconnected cells that form by incomplete cytokinesis and later undergo programmed breakdown. We recently identified similar cell clusters within the fetal mouse ovary, but the fate and functional significance of these germ cell cysts remained unclear. Here, we show that mouse cysts undergo programmed breakdown between 20.5-22.5 dpc, during which approximately 33% of the oocytes survive to form primordial follicles. This process accounts for most of the perinatal reduction in germ cell numbers and germ cell apoptosis reported by previous authors, and suggests that perinatal germ cell loss is a developmentally regulated process that is distinct from the follicular atresia that occurs during adult life. Our observations also suggest a novel function for a transient cyst stage of germ cell development. Prior to breakdown, mitochondria and ER reorganize into perinuclear aggregates, and can be seen within the ring canals joining adjacent germ cells. Cysts may ensure that oocytes destined to form primordial follicles acquire populations of functional mitochondria, through an active process that has been evolutionarily conserved. PMID- 11397005 TI - Assembly of trunk and limb blood vessels involves extensive migration and vasculogenesis of somite-derived angioblasts. AB - Vascular development requires the assembly of precursor cells into blood vessels, but how embryonic vessels are assembled is not well understood. To determine how vascular cells migrate and assemble into vessels of the trunk and limb, marked somite-derived angioblasts were followed in developing embryos. Injection of avian somites with the cell-tracker DiI showed that somite-derived angioblasts in unperturbed embryos migrated extensively and contributed to trunk and limb vessels. Mouse-avian chimeras with mouse presomitic mesoderm grafts had graft derived endothelial cells in blood vessels at significant distances from the graft, indicating that mouse angioblasts migrated extensively in avian hosts. Mouse graft-derived endothelial cells were consistently found in trunk vessels, such as the perineural vascular plexus, the cardinal vein, and presumptive intersomitic vessels, as well as in vessels of the limb and kidney rudiment. This reproducible pattern of graft colonization suggests that avian vascular patterning cues for trunk and limb vessels are recognized by mammalian somitic angioblasts. Mouse-quail chimeras stained with both the quail vascular marker QH1 and the mouse vascular marker PECAM-1 had finely chimeric vessels, with graft derived mouse cells interdigitated with quail vascular cells in most vascular beds colonized by graft cells. Thus, diverse trunk and limb blood vessels have endothelial cells that developed from migratory somitic angioblasts, and assembly of these vessels is likely to have a large vasculogenic component. PMID- 11397006 TI - DBHR, a gene with homology to dopamine beta-hydroxylase, is expressed in the neural crest throughout early development. AB - In a screen for genes involved in neural crest development, we identified DBHR (DBH-Related), a putative monooxygenase with low homology to dopamine beta hydroxylase (DBH). Here, we describe novel expression patterns for DBHR in the developing embryo and particularly the neural crest. DBHR is an early marker for prospective neural crest, with earliest expression at the neural plate border where neural crest is induced. Furthermore, DBHR expression persists in migrating neural crest and in many, though not all, crest derivatives. DBHR is also expressed in the myotome, from the earliest stages of its formation, and in distinct regions of the neural tube, including even-numbered rhombomeres of the hindbrain. In order to investigate the signals that regulate its segmented pattern in the hindbrain, we microsurgically rotated the rostrocaudal positions of rhombomeres 3/4. Despite their ectopic position, both rhombomeres continued to express DBHR at the level appropriate for their original location, indicating that DBHR is regulated autonomously within rhombomeres. We conclude that DBHR is a divergent member of a growing family of DBH-related genes; thus, DBHR represents a completely new type of neural crest marker, expressed throughout the development of the neural crest, with possible functions in cell-cell signaling. PMID- 11397007 TI - Chick CFC controls Lefty1 expression in the embryonic midline and nodal expression in the lateral plate. AB - Members of the EGF-CFC family of proteins have recently been implicated as essential cofactors for Nodal signaling. Here we report the isolation of chick CFC and describe its expression pattern, which appears to be similar to Cfc1 in mouse. During early gastrulation, chick CFC was asymmetrically expressed on the left side of Hensen's node as well as in the emerging notochord, prechordal plate, and lateral plate mesoderm. Subsequently, its expression became confined to the heart fields, notochord, and posterior mesoderm. Implantation experiments suggest that chick CFC expression in the lateral plate mesoderm is dependent on BMP signaling, while in the midline its expression depends on an Activin-like signal. The asymmetric expression domain within Hensen's node was not affected by application of FGF8, Noggin, or Shh antibody. Implantation of cells expressing human or mouse CFC2, or chick CFC on the right side of Hensen's node randomized heart looping without affecting expression of genes involved in left-right axis formation, including SnR, Nodal, Car, or Pitx2. Application of antisense oligodeoxynucleotides to the midline of Hamburger-Hamilton stage 4-5 embryos also randomized heart looping, but in contrast to the overexpression experiments, antisense oligodeoxynucleotide treatment resulted in bilateral expression of Nodal, Car, Pitx2, and NKX3.2, whereas Lefty1 expression in the midline was transiently lost. Application of the antisense oligodeoxynucleotides to the lateral plate mesoderm abolished Nodal expression. Thus, chick CFC seems to have a dual function in left-right axis formation by maintaining Nodal expression in the lateral plate mesoderm and controlling expression of Lefty1 expression in the midline territory. PMID- 11397008 TI - Purkinje fibers of the avian heart express a myogenic transcription factor program distinct from cardiac and skeletal muscle. AB - A rhythmic heart beat is coordinated by conduction of pacemaking impulses through the cardiac conduction system. Cells of the conduction system, including Purkinje fibers, terminally differentiate from a subset of cardiac muscle cells that respond to signals from endocardial and coronary arterial cells. A vessel associated paracrine factor, endothelin, can induce embryonic heart muscle cells to differentiate into Purkinje fibers both in vivo and in vitro. During this phenotypic conversion, the conduction cells down-regulate genes characteristic of cardiac muscle and up-regulate subsets of genes typical of both skeletal muscle and neuronal cells. In the present study, we examined the expression of myogenic transcription factors associated with the switch of the gene expression program during terminal differentiation of heart muscle cells into Purkinje fibers. In situ hybridization analyses and immunohistochemistry of embryonic and adult hearts revealed that Purkinje fibers up-regulate skeletal and atrial muscle myosin heavy chains, connexin-42, and neurofilament protein. Concurrently, a cardiac muscle-specific myofibrillar protein, myosin-binding protein-C (cMyBP-C), is down-regulated. During this change in transcription, however, Purkinje fibers continue to express cardiac muscle transcription factors, such as Nkx2.5, GATA4, and MEF2C. Importantly, significantly higher levels of Nkx2.5 and GATA4 mRNAs were detected in Purkinje fibers as compared to ordinary heart muscle cells. No detectable difference was observed in MEF2C expression. In culture, endothelin induced Purkinje fibers from embryonic cardiac muscle cells dramatically down regulated cMyBP-C transcription, whereas expression of Nkx2.5 and GATA4 persisted. In addition, myoD, a skeletal muscle transcription factor, was up regulated in endothelin-induced Purkinje cells, while Myf5 and MRF4 transcripts were undetectable in these cells. These results show that during and after conversion from heart muscle cells, Purkinje fibers express a unique myogenic transcription factor program. The mechanism underlying down-regulation of cardiac muscle genes and up-regulation of skeletal muscle genes during conduction cell differentiation may be independent from the transcriptional control seen in ordinary cardiac and skeletal muscle cells. PMID- 11397009 TI - Expression of axolotl DAZL RNA, a marker of germ plasm: widespread maternal RNA and onset of expression in germ cells approaching the gonad. AB - How germ cell specification occurs remains a fundamental question in embryogenesis. The embryos of several model organisms contain germ cell determinants (germ plasm) that segregate to germ cell precursors. In other animals, including mice, germ cells form in response to regulative mechanisms during development. To investigate germ cell determination in urodeles, where germ plasm has never been conclusively identified, we cloned a DAZ-like sequence from axolotls, Axdazl. Axdazl is homologous to Xdazl, a component of Xenopus germ plasm found in the vegetal pole of oocytes and eggs. Axdazl RNA is not localized in axolotl oocytes, and, furthermore, these oocytes do not contain the mitochondrial cloud that localizes Xdazl and other germ plasm components in Xenopus. Maternal Axdazl RNA is inherited in the animal cap and equatorial region of early embryos. At gastrula, neurula, and tailbud stages, Axdazl RNA is widely distributed. Axdazl first shows cell-specific expression in primordial germ cells (PGCs) approaching the gonad at stage 40, when nuage (germ plasm) appears in PGCs. These results suggest that, in axolotls, germ plasm components are insufficient to specify germ cells. PMID- 11397010 TI - Ovulation triggers activation of Drosophila oocytes. AB - Drosophila melanogaster mature oocytes in ovaries are arrested at metaphase I of meiosis. Eggs that have reached the uterus have released this arrest. It was not known where in the female reproductive tract egg activation occurs and what triggers it. We investigated when and where the egg is activated in Drosophila in vivo and at what meiotic stage the egg is fertilized. We found that changes in the egg's envelope's permeability, one feature of activation, initiate during ovulation, even while most of the egg is still within the ovary. The egg becomes impermeable as it proceeds down the oviducts; the process is complete by the time the egg is in the uterus. Cross-linking of vitelline membrane protein sV23 also increases progressively as the egg moves through the oviducts and the uterus. Activation also triggers meiosis to resume before the egg reaches the uterus, such that the earliest eggs that reach the uterus are in anaphase I. We discuss models for Drosophila egg activation in vivo. PMID- 11397011 TI - Cyclin E and its associated cdk activity do not cycle during early embryogenesis of the sea Urchin. AB - Female sea urchins store their gametes as haploid eggs. The zygote enters S-phase 1 h after fertilization, initiating a series of cell cycles that lack gap phases. We have cloned cyclin E from the sea urchin Strongylocentrotus purpuratus. Cyclin E is synthesized during oogenesis, is present in the germinal vesicle, and is released into the egg cytoplasm at oocyte maturation. Cyclin E synthesis is activated at fertilization, although there is no increase in cyclin E protein levels due to continuous turnover of the protein. Cyclin E protein levels decline in morula embryos, while cyclin E mRNA levels remain high. After the blastula stage, cyclin E mRNA and protein levels are very low, and cyclin E expression is predominant only in cells that are actively dividing. These include cells in the left coelomic pouch, which forms the adult rudiment in the embryo. The cyclin E present in the egg is complexed with a protein kinase. Activity of the cyclin E/cdk2 changes little during the initial cell cycles. In particular, cyclin E cdk2 levels remain high during both S-phase and mitosis. Our results suggest that progression through the early embryonic cell cycles in the sea urchin does not require fluctuations in cyclin E kinase activity. PMID- 11397012 TI - Tantalus, a novel ASX-interacting protein with tissue-specific functions. AB - The Drosophila trithorax- and Polycomb-group (trxG and PcG) proteins maintain activated and repressed transcriptional states at specific target gene loci. The Additional sex combs (Asx) gene is of particular interest as it appears to function in both protein complexes and yet its effects on target genes are more restricted. A novel protein, Tantalus (TAN), was identified in a yeast two-hybrid screen for ASX-interacting proteins that might confer tissue-specific ASX functions. TAN contains consensus nuclear localization sites and binds DNA in vitro. However, its subcellular localization varies in a tissue-specific fashion. In salivary glands, TAN is predominantly nuclear and associates with 66 euchromatic sites on polytene chromosomes, more than half of which overlap with ASX. These loci do not include the homeotic genes of the ANT and BX complexes bound by other PcG and trxG proteins. Rather, tan mutant defects are restricted to sensory organs. We show that one of these defects, shared by Asx, is genetically enhanced by Asx. Taken together, the data suggest that TAN is a tissue-specific cofactor for ASX, and that its activity may be partially controlled by subcellular trafficking. PMID- 11397013 TI - Mutation of the zebrafish glass onion locus causes early cell-nonautonomous loss of neuroepithelial integrity followed by severe neuronal patterning defects in the retina. AB - Mutation of the glass onion locus causes drastic neuronal patterning defects in the zebrafish retina and brain. The precise stratified appearance of the wild type retina is absent in the mutants. The glass onion phenotype is first visible shortly after the formation of optic primordia and is characterized by the rounding of cells and disruption of the ventricular surface in the eye and brain neuroepithelia. With exception of the dorsal- and ventral-most regions of the brain, neuroepithelial cells lose their integrity and begin to distribute ectopically. At later stages, the laminar patterning of retinal neurons is severely disrupted. Despite the lack of lamination, individual retinal cell classes differentiate in the glass onion retina. Mosaic analysis reveals that the glass onion mutation acts cell nonautonomously within the retina and brain, as neuroepithelial cell morphology and polarity in these tissues are normal when mutant cells develop in wild-type hosts. We conclude that the glass onion mutation affects cell-cell signaling event(s) involved in the maintenance of the neuroepithelial cell layer shortly after its formation. The disruption of neuroepithelial integrity may be the cause of the neuronal patterning defects following neurogenesis. In addition, the expression of the glass onion phenotype in a subset of neuroepithelial cells as well as its onset following the initial formation of the neuroepithelial sheets indicate the presence of genetically distinct temporal and spatial subdivisions in the development of this histologically uniform tissue. PMID- 11397014 TI - Morphogenesis of prechordal plate and notochord requires intact Eph/ephrin B signaling. AB - Eph receptors and their ligands, the ephrins, mediate cell-to-cell signals implicated in the regulation of cell migration processes during development. We report the molecular cloning and tissue distribution of zebrafish transmembrane ephrins that represent all known members of the mammalian class B ephrin family. The degree of homology among predicted ephrin B sequences suggests that, similar to their mammalian counterparts, zebrafish B-ephrins can also bind promiscuously to several Eph receptors. The dynamic expression patterns for each zebrafish B ephrin support the idea that these ligands are confined to interact with their receptors at the borders of their complementary expression domains. Zebrafish B ephrins are expressed as early as 30% epiboly and during gastrula stages: in the germ ring, shield, prechordal plate, and notochord. Ectopic overexpression of dominant-negative soluble ephrin B constructs yields reproducible defects in the morphology of the notochord and prechordal plate by the end of gastrulation. Notably disruption of Eph/ephrin B signaling does not completely destroy structures examined, suggesting that cell fate specification is not altered. Thus abnormal morphogenesis of the prechordal plate and the notochord is likely a consequence of a cell movement defect. Our observations suggest Eph/ephrin B signaling plays an essential role in regulating cell movements during gastrulation. PMID- 11397015 TI - Overexpression of camello, a member of a novel protein family, reduces blastomere adhesion and inhibits gastrulation in Xenopus laevis. AB - Vertebrate gastrulation involves complex coordinated movements of cells and cell layers to establish the axial structures and the general body plan. Adhesion molecules and the components of extracellular matrix were shown to be involved in this process. However, other participating molecules and detailed mechanisms of the control of gastrulation movements remain largely unknown. Here, we describe a novel Xenopus gene camello (Xcml) which is expressed in the suprablastoporal zone of gastrulating embryos. Injection of Xcml RNA into dorsovegetal blastomeres retards or inhibits gastrulation movements. Database searches revealed a family of mammalian mRNAs encoding polypeptides highly similar to Xcml protein. Characteristic features of the camello family include the presence of the central hydrophobic domain and the N-acetyltransferase consensus motifs in the C-terminal part, as well as functional similarity to Xcml revealed by overexpression studies in Xenopus embryos. Xcml expression results in the decrease of cell adhesion as demonstrated by the microscopic analysis and the blastomere aggregation assay. Cell fractionation and confocal microscopy data suggest that Xcml protein is localized in the secretory pathway. We propose that Xcml may fine tune the gastrulation movements by modifying the cell surface and possibly extracellular matrix proteins passing through the secretory pathway. PMID- 11397016 TI - CHAMP, a novel cardiac-specific helicase regulated by MEF2C. AB - MEF2C is a MADS-box transcription factor required for cardiac myogenesis and morphogenesis. In MEF2C mutant mouse embryos, heart development arrests at the looping stage (embryonic day 9.0), the future right ventricular chamber fails to form, and cardiomyocyte differentiation is disrupted. To identify genes regulated by MEF2C in the developing heart, we performed differential array analysis coupled with subtractive cloning using RNA from heart tubes of wild-type and MEF2C-null embryos. Here, we describe a novel MEF2C-dependent gene that encodes a cardiac-restricted protein, called CHAMP (cardiac helicase activated by MEF2 protein), that contains seven conserved motifs characteristic of helicases involved in RNA processing, DNA replication, and transcription. During mouse embryogenesis, CHAMP expression commences in the linear heart tube at embryonic day 8.0, shortly after initiation of MEF2C expression in the cardiogenic region. Thereafter, CHAMP is expressed specifically in embryonic and postnatal cardiomyocytes. At the trabeculation stage of heart development, CHAMP expression is highest in the trabecular region in which cardiomyocytes have exited the cell cycle and is lowest in the proliferative compact zone. These findings suggest that CHAMP acts downstream of MEF2C in a cardiac-specific regulatory pathway for RNA processing and/or transcriptional control. PMID- 11397017 TI - Nuclear plasticity and timing mechanisms of the initiation of alkaline phosphatase expression in cytoplasm-transferred blastomeres of ascidians. AB - Egg cytoplasm containing endoderm determinants was transferred to presumptive muscle or presumptive-epidermis blastomeres isolated from cleavage-stage embryos of the ascidian Halocynthia roretzi. We investigated three aspects of the expression of endoderm-specific alkaline phosphatase (ALP) activity. First, we examined whether ectopic ALP expression, an indication of ectopic endoderm formation, was promoted in cytoplasm-transferred blastomeres isolated at late cleavage stage. The results showed that the cell fate was converted by the introduced cytoplasm, even in recipient blastomeres in which the cell fate was already restricted to muscle or epidermis, and in those where expression of the muscle- or epidermis-specific genes was already initiated. Next, we examined the formation of endoderm and other tissue in embryos by double staining for ALP and muscle- or epidermis-specific marker. Regions positive for ALP and positive for muscle or epidermis marker were mutually exclusive. These results suggested that muscle- or epidermis-specific genes that were already expressed in the recipient blastomeres were down-regulated in ectopically forming endoderm cells. This is evidence for nuclear plasticity during ascidian embryogenesis. In the last series of experiments, we investigated the timing of the appearance of ALP activity in cytoplasm-transferred embryos. In the partial embryos that were derived from various combination of recipient blastomeres and donor cytoplasm obtained from various staged eggs and embryos, the timing seemed to coincide with the time that starts when cell fusion for cytoplasmic transfer was done. Therefore, the clock that determines the timing of the initiation of ALP expression is likely to start at the moment of cell fusion. Several possible hypotheses for the timing mechanism are discussed. PMID- 11397018 TI - Functional and regulatory analysis of the dictyostelium G-box binding factor. AB - The Dictyostelium discoidium G-box binding factor (GBF) is required for the induction of known postaggregative and cell-type-specific genes. gbf-null cells undergo developmental arrest at the loose-mound stage due to the absence of GBF targeted gene transcription. GBF-mediated gene expression is activated by stimulation of cell-surface, seven-span cAMP receptors, but this activation is independent of heterotrimeric G-proteins. To further characterize GBF, we assayed a series of GBF mutants for their ability to bind a G-box in vitro and to complement the gbf-null phenotype. In vitro DNA-binding activity resides in the central portion of the protein, which contains two predicted zinc fingers. However, in vivo GBF function requires only one intact zinc finger. In addition, expression of some GBF mutants results in a partial complementation phenotype, suggesting that these mutants are hypomorphic alleles. We used a 2.4-kb GBF promoter fragment to examine the regulation of GBF expression. GBF promoter reporter studies confirmed the previous finding that GBF transcription is induced by continuous, micromolar extracellular cAMP. We also show that, like the activation of GBF-regulated transcription, the induction of GBF expression requires cell-surface cAMP receptors, but not heterotrimeric G-proteins. Finally, reporter studies demonstrated that induction of GBF-promoter-regulated expression does not require the presence of GBF protein, indicating that GBF expression is not regulated by a positive autoregulatory loop. PMID- 11397019 TI - A model for national outcome audit in vascular surgery. AB - OBJECTIVE: The aim was to model vascular surgical outcome in a national study using POSSUM scoring. METHODS: One hundred and twenty-one British and Irish surgeons completed data questionnaires on patients undergoing arterial surgery under their care (mean 12 patients, range 1-49) in May/June 1998. A total of 1480 completed data records were available for logistic regression analysis using P POSSUM methodology. Information collected included all POSSUM data items plus other factors thought to have a significant bearing on patient outcome: "extra items". The main outcome measures were death and major postoperative complications. The data were checked and inconsistent records were excluded. The remaining 1313 were divided into two sets for analysis. The first "training" set was used to obtain logistic regression models that were applied prospectively to the second "test" dataset. RESULTS: using POSSUM data items alone, it was possible to predict both mortality and morbidity after vascular reconstruction using P-POSSUM analysis. The addition of the "extra items" found significant in regression analysis did not significantly improve the accuracy of prediction. It was possible to predict both mortality and morbidity derived from the preoperative physiology components of the POSSUM data items alone. CONCLUSION: this study has shown that P-POSSUM methodology can be used to predict outcome after arterial surgery across a range of surgeons in different hospitals and could form the basis of a national outcome audit. It was also possible to obtain accurate models for both mortality and major morbidity from the POSSUM physiology scores alone. PMID- 11397020 TI - Stroke from carotid endarterectomy: when and how to reduce perioperative stroke rate? AB - OBJECTIVES: To analyse four years of CEA with respect to the underlying mechanisms of perioperative stroke and the role of intraoperative monitoring in the prevention of stroke. PATIENTS AND METHODS: From January 1996 through December 1999, 599 CEAs were performed in 404 men and 195 women (mean age: 65 years, range: 39-88). All operations were performed under general anaesthesia using computerised electroencephalography (EEG) and transcranial Doppler (TCD). Any new or any extension of an existing focal cerebral deficit, as well as stroke related death were registered. Perioperative strokes were classified by time of onset (intraoperative or postoperative), outcome (minor or major stroke), and side (ipsilateral or contralateral). Stroke aetiology was assessed intraoperatively by means of EEG, TCD, completion arteriography or immediate re exploration, and postoperatively by duplex sonography, computerised tomography (CT) or magnetic resonance imaging (MRI) of the head. RESULTS: Perioperative stroke or death occurred in 20 (3.3%) patients. In four operations stroke was apparent immediately after surgery. Mechanisms of these strokes were ipsilateral carotid artery occlusion (1) and embolisation (3). In 16 patients stroke developed after a symptom-free interval (2-72 h, mean 18 h) due to occlusion of the internal carotid artery on the side of surgery (9). Other mechanisms were: contralateral occlusion of the internal carotid artery (1), postoperative hyperperfusion syndrome (1), intracerebral haemorrhage (1), and contralateral ischaemia due to prolonged clamping (1). In three procedures the cause was unknown. CONCLUSIONS: In our experience most strokes from CEA developed after a symptom-free interval and mainly due to thromboembolism of the operated artery. We suggest the introduction of additional TCD monitoring during the immediate postoperative phase. PMID- 11397021 TI - Prevention of postoperative wound haematomas and hyperperfusion following carotid endarterectomy. AB - OBJECTIVE: To study the incidence of wound haematomas and hyperperfusion following carotid endarterectomy and the effect of changes in perioperative management. METHODS: We undertook a prospective audit of the postoperative outcome of 300 consecutive carotid endarterectomies performed for a symptomatic stenosis of the internal carotid artery, under the care of a single consultant. RESULTS: audit of the first 100 operations between 1990-93 resulted in 4 changes to clinical practice. These included the use of Dacron instead of vein because of 3 vein patch blowouts, invasive postoperative monitoring of blood pressure, and the use of intravenous beta-blockers to control hypertension, because of 4 hyperperfusion injuries. The use of 10F suction drains was discontinued, because they did not prevent 8 wound haematomas. The results of the second 100 cases between 1994-97 and the third 100 cases between 1998-2000 confirmed no further hyperperfusion injuries or patch blowouts (p =0.01 and 0.04 respectively). Larger 14F suction drains were reintroduced for the third series because of thirteen haematomas in the second series (p =0.09). Only 4 haematomas occurred in the third series ( p =0.05). The need for beta-blockers fell in the third series due to the introduction of local anaesthesia (p =0.0001). CONCLUSION: The use of Dacron patches and postoperative control of hypertension has reduced the incidence of haemorrhage and hyperperfusion after carotid endarterectomy. Larger suction drains may also help. PMID- 11397022 TI - Immunophenotypic characterisation of carotid plaque: increased amount of inflammatory cells as an independent predictor for ischaemic symptoms. AB - OBJECTIVES: To investigate the inflammatory response within intact carotid plaques from carotid eversion endarterectomy (CEE) to determine the relationship between immunohistological plaque morphology and ischaemic cerebrovascular symptoms. MATERIAL AND METHODS: Intact CEE plaques from 71 patients with high grade (>70%) stenosis undergoing CEE (group I, symptomatic, n=42; group II, asymptomatic, n =29) and 12 normal postmortem arteries (control group) were analysed with specific antibodies to inflammatory cells (T-Lymphocytes (CD3, CD4), cytotoxic T-cells (CD8), B-lymphocytes (CD20), natural killer cells (CD57), macrophages (CD68)), endothelial adhesion molecules (ICAM-1 (CD54), P-selectin (CD62P), E-selectin (CD62E), VCAM-1 (CD106) and T-lymphocyte co-stimulatory molecule (CD40)) and procoagulatory modulators (thrombomodulin (CD141), tissue factor (CD142)). Both groups were matched for gender, age, risk factors, degree of carotid artery stenosis. Plaques were measured using a semiquantitative score system in a blinded fashion by two observers. Statistical analysis of the group differences were performed by using the Kruskal-Wallis test and the Multitest Procedure with Permutation-Testing. Significance was taken as a p<0.05. RESULTS: There were significantly more inflammatory cells, an overexpression of P-selectin and the procoagulatory markers thrombomodulin and tissue factor in symptomatic compared to both asymptomatic plaques and the ones of the control group. In both groups there was no significance for ICAM-1, VCAM-1, macrophages and co stimulatory molecule CD40. There was also no significance for any factor between the asymptomatic and the control group. However, the differences between the symptomatic and the asymptomatic group were highly significant for all factors. CONCLUSION: These data suggest that structural changes and inflammatory damage within the individual plaque seems to be a critical step in promoting plaque rupture with embolic sequelae. PMID- 11397023 TI - Asymptomatic leg atherosclerosis is reduced by regular physical activity. Longitudinal results from the cohort "men born in 1914". AB - OBJECTIVE: To study whether physical activity is associated with reduced occurrence of asymptomatic leg atherosclerosis. DESIGN: Longitudinal and cross sectional analyses of the population-based cohort "men born in 1914". METHODS: Comparison of the systolic ankle/arm pressure index (AAI) at age 68 in groups who were sedentary, performed some activity, and performed regular physical training at 55 and 68 years of age. RESULTS: At 55 years of age, 100 (27%) were sedentary, 209 (58%) reported some physical activity and 54 (15%) reported regular physical training. At 68 years, 194 men (53%) reported the same degree of physical activity, 127 (35%) reported a higher physical activity, and 42 (12%) reported lower physical activity. Physical activity at 55 years (p =0.03) and increased physical activity between 55 and 68 years (p =0.03) were both associated with higher AAI at 68 after adjusting for potential confounders. At 68 years, AAI was 0.89+/-0.21, 1.01+/-0.13 and 1.05+/-0.11, respectively, in men who were sedentary, reported some physical activity, and regular physical training (p =0.0002). This association remained significant after adjustments for potential confounders. CONCLUSIONS: regular physical activity is associated with reduced occurrence of asymptomatic leg atherosclerosis, even in men taking up exercise after age of 55. PMID- 11397024 TI - The bridge graft: a new concept for infrapopliteal surgery. AB - BACKGROUND: The long-term results of ePTFE grafts are particularly poor in crural reconstructions. We report on a novel surgical technique, whereby both run-off and anastomotic mismatches are concomitantly addressed. PATIENTS AND METHODS: Short segments of vein grafts (5-15 cm in length) were used to bridge two crural artery segments. Subsequently, a femoro-distal ePTFE graft was anastomosed to the bridge graft. Venous valves were made incompetent to allow bi-directional flow. In a retrospective series of 45 patients with crural bridge grafts, 12 patients were in stage III and 33 in stage IV. In 18 patients the reconstruction was the first procedure and in the remaining 28 patients it was the first or second re operation. RESULTS: The primary patency rate at 1, 2, 3 and 4 years was 53, 44, 35 and 26% respectively. The secondary patency rate was 67, 53, 49 and 39% respectively. The corresponding limb salvage rate was 70, 61, 56 and 45%. In a small subgroup of patients, in which the crural bridge was the first reconstructive procedure, the primary patency was 76 at 1 year and 64 at 4 years. CONCLUSION: convincing long-term crural bridge grafts should be considered in those patients who have more than one crural or pedal artery available for grafting and an insufficient length of saphenous vein. PMID- 11397025 TI - Intraoperative endothelial damage is associated with increased risk of stenoses in infrainguinal vein grafts. AB - OBJECTIVES: Saphenous vein injury is believed to predispose to vein graft stenoses. The aims of this study were to assess the endothelial injury associated with infrainguinal vein bypass surgery by serial measurements of serum thrombomodulin concentration as well as by platelet scintigraphy, and to relate these findings with the postoperative development of stenoses. METHODS: In 35 patients undergoing vein bypass surgery serum thrombomodulin concentration was measured pre- and postoperatively. Autologous 111-indium labelled platelets were administered into the common femoral artery immediately after restoration of flow in the graft and scintigraphic images were obtained 4 and/or 24 h later. RESULTS: Serum thrombomodulin increased markedly from median 17 ng/ml preoperatively to 32 ng/ml 1 day after surgery (p =0.00002). Platelet scintigraphy revealed a total of 62 focal activity accumulations, the majority being located in the anastomotic regions. Among the 30 patients with grafts remaining patent at 30 days stenoses occurred in nine (16%) of 55 regions with scintigraphic platelet accumulations as compared to only four (4%) of 94 regions without platelet accumulations (p =0.03). CONCLUSIONS: The very high predictive value of a negative platelet scintigraphy (96%) strongly suggests that localised perioperative endothelial injury is an important pathogenetic factor in the development of vein graft stenoses. PMID- 11397026 TI - Harnessing haemodynamic forces for the suppression of anastomotic intimal hyperplasia: the rationale for precuffed grafts. AB - OBJECTIVES: Standardisation of cuff geometry by manufacturing prosthetic precuffed grafts (PCG) theoretically optimises haemodynamic forces. This study was designed in order to determine whether these beneficial flow patterns are replicated in vivo in PCG. PATIENTS AND METHODS: Flow visualisation and Doppler studies performed on anatomically accurate PCG models characterised in vitro anastomotic flow patterns. Thirty-two patients (median age 68 years) in whom autologous vein was unavailable, underwent bypass using PCG. Post-operative analysis included qualitative assessment of flow within the distal anastomosis using Doppler colour flow mapping. Cardiac gating techniques and assessment of velocity distribution were performed to gain additional information. These in vivo results were validated against the bench studies. RESULTS: A cohesive vortex was identified within the distal anastomosis of in vitro models and had an integral relationship with the cardiac cycle. This flow structure was also characterised using Doppler colour flow mapping in both longitudinal and transverse planes, confirming the location of the vortex within the body and proximal part of the anastomosis. Twenty-two patients (69%) undergoing bypass with a PCG underwent successful Doppler assessment one week post-operatively, of whom 17 (77%) had a vortical flow structure identified at the distal anastomosis, similar to that characterised in vitro. Cardiac gating verified the same integral relationship of the vortex with the cardiac cycle as that described in vitro. CONCLUSION: The geometric configuration of precuffed grafts induced vortices within the distal anastomoses in 17 out of 22 patients undergoing arterial reconstruction, thereby harnessing the haemodynamic forces that may suppress anastomotic hyperplasia and improve patency rates. PMID- 11397027 TI - Myocardial injury and systemic fibrinolysis in patients undergoing repair of ruptured abdominal aortic aneurysm: a preliminary report. AB - BACKGROUND: ruptured abdominal aortic aneurysm (AAA) is associated with inhibition of systemic fibrinolysis. Hypofibrinolysis is a risk factor for ischaemic myocardial injury, one of the commonest complications of ruptured AAA repair. Cardiac troponin I (cTnI) is one of the most sensitive and specific marker of myocardial injury currently available. OBJECTIVE: To examine, for the first time, the relationship between fibrinolytic activity and myocardial injury in patients operated for ruptured AAA. METHODS: Twenty patients (18 men and 2 women of median age 74, range 65-86 years) undergoing repair of ruptured AAA were prospectively studied. Plasma tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1) activity were measured pre-operatively, immediately before and five minutes following aortic clamp release. Serum cTnI was measured pre-operatively, 6 and 24 h following clamp release. Results cTnI was detectable at one or more sample points in 13 (65%) patients, and in 7 out of 8 patients who suffered major cardiac complications. There was a significant negative correlation between pre-operative t-PA activity and cTnI before operation (r = 0.55, p = 0.01) and 6 h ( r =-0.51, p =0.02) after clamp release. There was a significant positive correlation between pre-operative PAI activity and cTnI before operation (r =+0.50, p =0.03), 6 h ( r =+0.47, p =0.04) and 24 h ( r =+0.50, p =0.03) after clamp release. There was no correlation between pre- and intra-operative hypotension or blood transfusion requirement and cTnI release. CONCLUSIONS: Hypofibrinolysis during ruptured AAA repair is associated with the development of peri-operative myocardial injury. The causal mechanisms underlying this state are not clear but treatment of this prothrombotic/hypofibrinolytic diathesis may help to limit myocardial cell necrosis. PMID- 11397028 TI - The long-term benefits of a single scan for abdominal aortic aneurysm (AAA) at age 65. AB - AIM: To investigate the efficacy of a single ultrasonic scan at age 65 to identify patients at risk from ruptured abdominal aortic aneurysm (AAA). METHOD: A total of 6058 men aged 64-81 were recruited to a randomised trial, and 3000 were invited to attend a single screening test. An additional population of 1011 men was offered screening as they reached age 65. If a normal aorta was identified in this sub-group, further scans were offered at two-yearly intervals. Follow up and treatment of those identified as having an aortic dilatation of 3 cm or greater was undertaken. All subject groups were monitored for deaths occurring over the study period, and date and cause of death were recorded. RESULTS: A total of 2212 men attended screening in the randomised trial; the overall compliance was 74%, and prevalence of AAA was 7.7%. Compliance decreased, and prevalence increased, with age. Mortality from ruptured AAA was reduced by 68% at 5 years (screened group compared to the age-matched control population), and by 42% in the study arm (screened and refusers) compared with controls. The benefit persisted at ten years (53% and 21% respectively). Of the uncontrolled sample of 1011 men offered a scan at age 65, 681 attended and 649 of these were found to have a normal aortic diameter; re-screening demonstrated new aneurysm development in 4% over ten years. The aortic diameters of the new AAAs were under 4 cm and would therefore have a low risk of rupture.1 Mortality from rupture in all those with an initially normal aortic diameter was low, at 1 case per 1000 scans over ten years. CONCLUSION: Screening once for AAA at age 65 can identify the majority of AAA that are of clinical significance and can identify a large population at low risk from rupture who do not require surveillance. This policy has been effective when combined with selective treatment in reducing the risk of rupture for ten years in those who attend the screening programme. PMID- 11397029 TI - All sealed endoleaks are not the same: a treatment strategy based on an ex-vivo analysis. AB - PURPOSE: factors contributing to pressure transmission through thrombosed or sealed endoleaks have not been elucidated. The purpose of this investigation was to create an ex-vivo model that mimics patent and sealed endoleaks and that can quantitatively analyse the effects of length, diameter and thrombus on pressure transmission to the interior of the aneurysm sac. METHODS: In the ex-vivo model, endoleak channels (ELCs) of various lengths (2 cm, 6 cm, 10 cm) and diameters (0.6 cm, 1.0 cm, 1.4 cm) were constructed using polytetrafluoroethylene (PTFE) grafts and attached to an artificial aneurysm sac. These ELCs were incorporated within a mock circulation made of rubber tubing connected to a pulsatile pump. Peak systolic pressure (PSP) was recorded in the aneurysm sac, distal to each ELC. Subsequently the ELCs were filled with human thrombus, and the pressure measurements repeated (n =5). Data was evaluated by regression analysis. RESULTS: Peak systolic pressure in the artificial circulation was maintained at 150 mmHg. In the absence of thrombus pressure did not change across the ELC, regardless of its length or diameter. In the presence of organised thrombus, the pressure curves distal to the ELC were dampened, and the pressure reduction was directly proportional to the length and inversely proportional to the diameter of the ELC. Regression analysis indicated statistical significance. CONCLUSION: In the absence of thrombosis, pressure transmitted via an ELC to the aneurysm sac is unchanged regardless of its length or diameter. All sealed endoleaks also transmit pressure. However, when an endoleak has thrombosed, pressure reduction is directly proportional to the length and inversely proportional to the diameter of its channel. This ex-vivo model suggests that Type 2 endoleaks with longer channels and smaller diameters would derive a greater benefit from adjunctive manoeuvres (coil embolisation) that hasten thrombosis. On the other hand, thrombosis of endoleaks with short and wide channels (e.g. Type 1) may not result in substantial pressure reduction within the aneurysm sac and a successful outcome. PMID- 11397030 TI - Long saphenous vein stripping and quality of life--a randomised trial. AB - OBJECTIVES: To assess the quality of life of patients undergoing sapheno-femoral junction (SFJ) ligation and long saphenous vein stripping (LSV), using two different techniques. DESIGN: Prospective, randomised trial. MATERIALS AND METHODS: Eighty patients were recruited and randomised to either Perforate Invagination (PIN) stripping (43) or Conventional stripping (37). Patients completed the Short Form 36 (SF-36) and EuroQol (EQ) questionnaires preoperatively, and postoperatively at 6 weeks and 6 months. RESULTS: Bodily pain, role function and physical summary were significantly improved at 6 months in the PIN stripping group. In the Conventional group, bodily pain and physical function were similarly improved, but not role function. EQ global quality of life was significantly and progressively improved at 6 weeks and 6 months in the PIN group (global score p<0.003; self-rated score p <0.001). In the Conventional group there was no overall improvement in global score or self-rated health. CONCLUSIONS: Primary varicose vein surgery is associated with significant and progressive improvements in quality of life scores. Whilst overall quality of health does improve in the Conventional group, this appears to be to a lesser extent than in the PIN group. PMID- 11397031 TI - Non-saphenofemoral venous reflux in the groin in patients with varicose veins. AB - OBJECTIVES: To investigate the incidence, clinical significance, anatomical variation and physiology of non-saphenofemoral venous reflux (non-SF reflux) in the groin. DESIGN: Prospective study. MATERIALS: A total of 1072 vascular diagnostic workups in 680 patients with possible venous diseases to the legs were included. METHODS: Duplex scanning and air plethysmography. RESULTS: A total of 1022 legs had venous diseases. Of these, 101 (9.9%) had non-SF reflux in the groin. Such reflux occurred in recurrent varicose veins (RVV) in 16.3%, in primary varicose veins (PVV) in 6.1% and in deep venous thrombosis (DVT) in 8.0%. Two patterns of reflux were distinguished: epigastric reflux from lower abdominal wall veins (71 legs) and pudendal reflux from perineal and/or gluteal veins (30 legs). Pudendal reflux was almost exclusive to women and did not occur with DVT. If there was only non-SF reflux at the groin the venous filling indices (VFI) were close to normal (1.7+/-1.0 ml/s for RVV, 1.9+/-1.2 for PVV, 1.7+/-1.0 for DVT) and no active ulcers were observed. However, if non-SF reflux was associated with saphenofemoral or other reflux the VFIs (3.3+/-2.3 ml/s for RVV, 3.8+/-1.5 ml/s for PVV) were abnormal (p <0.05) and ulcers occurred in 11/32. CONCLUSION: Non-SF reflux in the groin is common. Such reflux may be missed at initial surgery and lead to recurrence of varicose veins. However, the venous physiological disturbance of such reflux is mild and it is not associated with ulcers unless combined with reflux at other sites in the leg. PMID- 11397032 TI - A survey of deep venous thrombosis management by consultant vascular surgeons in the United Kingdom and Ireland. AB - OBJECTIVES: The aim of this study was to detail the current consensus amongst vascular surgeons in Great Britain and Ireland regarding their investigation and management of patients with suspected or proven deep vein thrombosis (DVT). METHODS: The database of the Vascular Surgical Society of Great Britain and Ireland (VSS) was utilised to send coded postal questionnaires to all consultant surgeon members. RESULTS: Replies were received from 281 (65%) consultants. Duplex ultrasound is used alone to confirm DVT by 69% of respondents. A thrombophilia screen is always performed by 14% of consultants, for patients with proven DVT, and is more commonly requested by consultants based in a teaching hospital. The majority (57%) of consultants treat DVT with unfractionated heparin (UFH) and warfarin, whereas only 38% utilise low molecular weight heparins (LMWH) and warfarin. A management policy for DVT is reported to be in place by 59%, and a set policy for the specific management of calf vein DVT by just 20%. CONCLUSION: New diagnostic modalities and treatments have been developed for DVT that are more convenient and cost-effective. Although clinical guidelines for the management of patients with DVT are beginning to emerge, there is still a wide discrepancy in many areas of DVT management, and practice at variance with the current evidence base, amongst vascular surgeons in the United Kingdom and Ireland. PMID- 11397033 TI - Arterial graft occlusion after pelvic surgery in the lithotomy position. PMID- 11397034 TI - Progression of peri-aortic fibrosis despite endovascular repair of an inflammatory aneurysm. PMID- 11397035 TI - Spontaneous ruptured lumbar artery in a chronic renal failure patient. PMID- 11397036 TI - Parallel stent placement for maldeployed iliac stent. PMID- 11397037 TI - Bilateral one-stage carotid endarterectomy--Is there an indication? PMID- 11397038 TI - A rare case of mesenteric infarction. PMID- 11397041 TI - Native and Hydrophobically Modified Human Immunoglobulin G at the Air/Water Interface. AB - The adsorption of human immunoglobulin G (IgG) at the air/water interface was monitored both by the in situ radiotracer technique using [(14)C] labeled IgG and by surface tension measurements. The results reveal that adsorption of IgG from single protein systems displays bimodality due to molecular rearrangements at the interface. Above the threshold value of 1.5x10(-2) mg/ml solution concentration, adsorbed IgG molecules reoriented from the side-on to the end-on configuration. The existence of a lag time which did not appear in Gamma=f(t) curves, was observed in Pi=f(t) relationships at low protein concentrations and was due to the limits of the surface pressure technique to detect protein adsorption. The adsorption of native IgG was also carried out in the presence of a hydrophobized IgG obtained by grafting capryloyl residues to its lysine groups by reaction with N-hydroxysuccinimide ester of caprylic acid, which yielded 19 covalently bound alkyl chains to the IgG molecule (19C(8)-IgG). This modified IgG exhibited enhanced adsorption at the air/water interface, as manifested by its increased adsorption efficiency relative to the native protein. Sequential and competitive adsorption experiments from binary mixtures of native IgG and 19C(8)-IgG clearly demonstrate that the displacement of the native protein from the air/water interface strongly depended on the manner of how 19C(8)-IgG and native IgG competed with each other. When the two proteins competed simultaneously, 19C(8) IgG predominantly occupied the available area but when native IgG was adsorbed first, for 2 h, the sequentially adsorbed 19C(8)-IgG was incapable of substantially displacing it from the interface. Copyright 2001 Academic Press. PMID- 11397042 TI - Interrelations between Adsorption Energies and Local Isotherms, Local Monolayer Capacities, and Energy Distribution Functions, as Determined for Heterogeneous Surfaces by Inverse Gas Chromatography. AB - Physicochemical parameters for adsorption of gases at the submonolayer regions of heterogeneous solid surfaces are measured experimentally as a function of time, and then interrelated as local isotherms θ against adsorption energy varepsilon, fractional changes of adsorption sites f(varepsilon)/c*(max) against varepsilon, θ against f(varepsilon)/c*(max), and distribution functions θ f(varepsilon)/c*(max) over adsorption energy values varepsilon, without using at all the well-known integral equation Theta(p, T)=integral(infinity)(0)θ(p, T, varepsilon)f(varepsilon)dvarepsilon and assumptions concerning the pair f(varepsilon) and θ(p, T, varepsilon). The method uses only chromatographic experimental data obtained by the inverse gas chromatography technique known as reversed-flow gas chromatography. It has been applied to the adsorption of cis-2-butene and trans-2-butene onto particles of Penteli marble at temperatures of 302, 314, 323, and 333 K. The results obtained are comparable with those calculated on the basis of the well-known integral equation. Copyright 2001 Academic Press. PMID- 11397043 TI - Adsorption Behaviors of L-Histidine and DL-Tryptophan on Cholesterol, Silica, Alumina, and Graphite. AB - The adsorption of L-histidine (L-His) and DL-tryptophan (DL-Trp) on cholesterol, silica, alumina, and graphite surfaces from aqueous solution at pH 9.2 has been studied in the temperature range of 25 to 50 degrees C. The data have been analyzed, wherever applicable, by Langmuir and Hill equations to obtain (1) the monolayer adsorption capacity (x(m)) and the equilibrium constant (k') for the adsorption and desorption processes and (2) the Hill coefficient (n) at different temperatures. The results of adsorption on different kinds of adsorbents have been compared. The energetic parameters for the adsorption process have also been evaluated and compared. Copyright 2001 Academic Press. PMID- 11397044 TI - Interaction of a Zwitterionic Surfactant with Synthetic Clays in Aqueous Suspensions: A Multinuclear Magnetic Resonance Study. AB - (1)H and (13)C NMR signal assignment of laurylamidopropylbetain has been obtained from 2D COSY, HETCOR, and HMBC pulse sequences. (13)C and (14)N NMR relaxation data show interaction of the polar head of the surfactant with low-charged clays dispersed in aqueous solution. The sodium cation exchange rate between the solid surface and the bulk is strongly affected by the charge of the clay layer, as seen from (23)Na NMR data. Such an exchange is also observed with clay suspended in sodium dodecyl sulfate solutions. Copyright 2001 Academic Press. PMID- 11397045 TI - Determination of the Adsorbed Phase Volume and Its Application in Isotherm Modeling for the Adsorption of Supercritical Nitrogen on Activated Carbon. AB - Experimental excess adsorption isotherms of nitrogen on activated carbon were measured in the range of 103-298 K and pressures up to 10 MPa. A method for determining the volume of the adsorbed phase from the experimental data for the supercritical temperatures was proposed. Such a volume with a new isotherm equation was used in modeling experimental excess isotherms, and a satisfactory fit was achieved. Copyright 2001 Academic Press. PMID- 11397046 TI - Structural Characterization and Iron(III) Binding Ability of Dimeric and Polymeric Lignin Models. AB - To understand the complexation in solution and the sorption of iron(III) on soluble and solid fractions of lignin, a dimeric model (guaiacyl-beta guaiacylglycerol ether, called beta-O-4) and a polymeric model (dehydrogenation polymer resulting from polymerization of coniferyl alcohol) of lignin have been synthesized and characterized with chromatographic, solution, and solid state (13)C CP-MAS NMR and XPS spectroscopies. The beta-O-4 dimer is a monoacid (HL). Potentiometric studies in aqueous solution at 25 degrees C and 1 mol L(-1) ionic strength (KNO(3)) indicated formation of two stable complexes, FeL(2+) and probably FeL(OH)(+), which shows that the soluble fraction of lignin binds metals, indicating that they are transported by water through the soils. The binding of iron(III) on the DHP polymer was then investigated. The sorption experiments have shown a great affinity of iron for the solid with a maximum of adsorption since pH 5. A pulsed-ESR study has revealed surface oxidation by the iron(III) cation, which leads to iron(II) and semiquinonic radicals on the polymer surface, with a radical concentration of about 5x10(17) spin/g. Copyright 2001 Academic Press. PMID- 11397047 TI - Osmotic Pressure, Small-Angle X-Ray, and Dynamic Light Scattering Studies of Human Serum Albumin in Aqueous Solutions. AB - Osmotic pressure measurements of human serum albumin (HSA) dissolved in water and in 0.01, 0.1, and 1.0 M phosphate buffer are reported as a function of the protein concentration. Two different forms of the protein were studied: defatted HSA (HSA1) and HSA with fatty acids (HSA2). The measured values of the osmotic coefficient were well below 1, indicating large deviations from ideality even for dilute protein solutions. The measured values increased with increasing HSA concentration and the increase was a function of pH. For higher concentrations of added phosphate buffer, the pH of solution had less influence on the measured osmotic pressure. The osmotic pressure of HSA1 in water was found to be considerably lower than that of the HSA2 modification. This effect was ascribed to formation of dimers in the HSA1 solution. The osmotic measurements were complemented by the small-angle X-ray scattering (SAXS) and dynamic light scattering (DLS) studies of dilute HSA solutions in water. The SAXS and DLS data confirmed the dimerization of HSA1 molecules under these conditions. Detailed analysis of the SAXS data suggested a parallel orientation of two protein molecules in a dimer. Copyright 2001 Academic Press. PMID- 11397048 TI - Correlation and Prediction of Osmotic Pressures for Aqueous Bovine Serum Albumin NaCl Solutions Based on Two Yukawa Potentials. AB - A new equation of state is proposed to correlate and predict the osmotic pressure data for aqueous bovine serum albumin (BSA) solutions with different NaCl concentrations and pH values with only one adjustable parameter. The Carnahan Starling equation represents the contribution of the hard sphere repulsion to the osmotic pressure. The attractive dispersion and double-layer repulsion interactions are represented by two Yukawa potentials, respectively. The equation of state of Duh and Mier-Y-Teran for one Yukawa potential is expanded to two Yukawa potentials to describe the nonidealities of the charged BSA-aqueous NaCl solution, instead of the classical DLVO theory. The average relative deviation of correlation of the osmotic pressure in 0.15 M NaCl solution is 18%. The average relative deviation of prediction in 1-5 M NaCl solutions is 20.33%. A comparison with other models and the limitations of our model are discussed. Copyright 2001 Academic Press. PMID- 11397049 TI - Hydrophobic Laminin-Related Peptides: Synthesis and Surface Properties. AB - The synthesis of hydrophobic peptide derivatives related to the laminin sequence [YIGSR(NH(2))] is described. Hydrophobicity is achieved by the attachment of decanoic, myristic, or stearic acids to the amino terminal end of the peptide. Moreover, a cholesterol residue was also introduced as succinimidoyl-cholesteryl moiety at the same position. These peptidic compounds are designed to be inserted into lipid bilayers to prepare, what can be considered as, immunoliposomes to target these vesicles to tumor cells. Physicochemical aspects related to their surface activity, insertion into lipid layers, spreadibility, formation of aggregates, and haemolytic activity have been studied as a previous step in the selection of the most convenient derivative. The results obtained indicate that these peptide derivatives show a high tendency to form aggregates in aqueous media, this fact reducing their interaction with lipid mono- and bilayers. The most suitable derivatives for interacting with liposomes are myristoyl and decanoyl. Copyright 2001 Academic Press. PMID- 11397050 TI - Rhenium-Based Hydrosols: Preparation and Properties. AB - Transmission electron microscope (TEM), ultraviolet-visible (UV/vis), electrospray mass spectrometric (ESMS), and X-ray photoelectron spectroscopic (XPS) studies are presented on a little studied rhenium hydrosol system produced by reduction of aqueous K(2)ReCl(6) with hydrazine in the presence or absence of a gum arabic protecting agent. The studies indicate that hydrazine-generated "rhenium hydrosols" are unstable in water and slowly dissolve over time in aqueous media to form the highly stable perrhenate ion. Copyright 2001 Academic Press. PMID- 11397051 TI - Preparation of Nanosized Metallic Particles in Polyaniline. AB - The preparation of nanosized gold and palladium particles in polyaniline has been carried out via the reduction of AuCl(3) or Pd(NO(3))(2) by polyaniline in either aqueous media or N-methylpyrrolidinone (NMP). When the reduction of AuCl(3) was carried out in NMP solutions of polyaniline, the Au particles were on the order of 20 nm. The reduction of AuCl(3) or Pd(NO(3))(2) by polyaniline in the powder form in aqueous media resulted in the accumulation of the elemental Au or Pd on the surface of the polyaniline particles. Subsequent dissolution of the polyaniline in NMP resulted in metal particles of about 50 to 200 nm being dispersed in the NMP solution of polyaniline. The rate of metal salt reduction and the size of the metal particles were found to be strongly dependent on the medium used, the initial ratio of metal ions to polyaniline, and the reaction time. The polyaniline-metal particle systems were characterized using X-ray photoelectron spectroscopy, UV-visible absorption spectroscopy, and FTIR spectroscopy. Scanning and transmission electron microscopy and laser light scattering were used to determine the size of the metal particles in polyaniline. Copyright 2001 Academic Press. PMID- 11397052 TI - Analysis of Light Scattering Data on the Calcium Ion Sensitivity of Caseinate Solution Thermodynamics: Relationship to Emulsion Flocculation. AB - We describe the quantitative interrelation between the thermodynamic parameters of caseinate submicelles in the presence of calcium ions (0-14 mM) in aqueous medium and the capacity of the protein to induce depletion flocculation in oil-in water emulsions at pH 7.0 and ionic strength 0.05 mol dm(-3). Measurements have been made by static and dynamic multiangle laser light scattering of the weight average molecular weight, the radius of gyration, the hydrodynamic radius, and the second virial coefficient of caseinate submicelles in aqueous solution. Successive thermodynamic approximations with and without consideration of correlations between caseinate submicelles have been used to calculate the osmotic pressure in caseinate aqueous solutions and the free energy of the depletion interaction between droplets in oil-in-water emulsions stabilized by caseinate. Numerical results from both thermodynamic approximations are in reasonably good agreement with experiment, predicting a pronounced decrease in the strength of the depletion attraction at concentrations of Ca(2+) in the range 4-8 mM (with a minimum value at 8 mM). This correlates well with the great enhancement of stability of these emulsions with respect to flocculation in comparison with systems having no added ionic calcium and emulsions with lower (2 mM) or higher (10 mM) Ca(2+) contents. Nevertheless, the allowance for interactive correlations between caseinate submicelles seems to lead to a better prediction of emulsion flocculation on a qualitative level over the whole range of Ca(2+) concentrations studied (2-14 mM). The calculated pronounced decrease in depletion interaction strength is attributable to marked changes in weight average molecular weight and mean size of aggregates, and to more positive values of the second virial coefficient of caseinate submicelles with increasing Ca(2+) content. Finally, we discuss the part played by the electrical charge on the protein in determining the overall strength of the flocculation-inducing attractive interactions between droplets. Copyright 2001 Academic Press. PMID- 11397053 TI - Electroosmotic Flow in Microchannels. AB - The electroosmotic flow induced by an applied electrostatic potential field through microchannels between two parallel plates and a 90 degrees bend is analyzed in this work. A nonlinear, two-dimensional Poisson-Boltzmann equation governing the electrical double-layer field and the Laplace equation governing the electrostatic field distribution in microchannels are numerically solved using a finite-difference method. A body force caused by the interaction between the electrical double-layer field and the applied electrostatic field is included in the full Navier-Stokes equations. The effects of the electrical double-layer field and the applied electrostatic field on the fluid velocity distribution, pressure drop, and skin friction are discussed. A small pressure drop along the parallel plates is detected, although it is always neglected in the literature. Pressure is not a constant across the channel height. The axial velocity profile is no longer flat across the channel height when the Reynolds number is large. A separation bubble is detected near the 90 degrees junction when the Reynolds number is large. Copyright 2001 Academic Press. PMID- 11397054 TI - Direct Differentiation of the Types of Polarization Responsible for the Electrorheological Effect By a Dielectric Method. AB - Debye polarization, ionic displacement polarization, and Maxwell-Wagner (interfacial) polarization are discussed in this paper, because they would most likely take place in an electrorheological (ER) suspension. The temperature dependences of the dielectric loss tangent maximums governed by these three types of polarization are theoretically found to be quite different. Given this fact, a method that can directly distinguish the polarization type and clarify which polarization would be mainly responsible for the ER effect is proposed. Two kinds of typical ER suspensions, heterogeneous particle type and homogeneous liquid crystalline polymer type, are studied using our method. It is found that Maxwell Wagner polarization would be responsible for the ER effect both in a heterogeneous and in a homogeneous ER system. These findings present direct experimental evidence for the previous assumption that the Maxwell-Wagner polarization would dominate in the heterogeneous ER system and also shed light on the ER mechanism in a liquid crystalline polymer-type ER system. Copyright 2001 Academic Press. PMID- 11397055 TI - Electric Potential and Reaction Rates at Charged Surfaces in Asymmetric Electrolytes-An Analytic Approach. AB - The electric potential and reaction rates of ions hitting the chemically active surfaces of microcrystals in an asymmetric electrolyte are computed analytically. Following ideas of Debye we start by solving the Poisson-Boltzmann equations and by determining the electric potential of the transport equations. We find distinct deviations when comparing our result with the Gouy-Chapman formula. In a simple model approximating a situation in which lead and hydrogen ions can react at goethite surfaces we compute analytically the currents of ions diffusing to the surfaces of microcrystals where they undergo a chemical reaction. We compute the reaction rates that can be controlled either by chemical reactions at the surface of the microcrystals or by diffusional transport. For realistic parameters of our model we find that the diffusional transport is the rate determining step. Copyright 2001 Academic Press. PMID- 11397056 TI - The Modification of Hydroxyl Surfaces of Formamide-Intercalated Kaolinites Synthesized by Controlled Rate Thermal Analysis. AB - Controlled rate thermal analysis (CRTA) technology made possible the separation of adsorbed formamide from intercalated formamide in formamide-intercalated kaolinites. X-ray diffraction shows that the CRTA-treated formamide-intercalated kaolinites remain expanded after CRTA treatment. The Raman spectra of the CRTA treated formamide-intercalated kaolinites are significantly different from those of the intercalated kaolinites with both intercalated and adsorbed formamide. An intense band is observed at 3629 cm(-1), attributed to the inner surface hydroxyls hydrogen bonded to the formamide. Broad bands are observed at 3600 and 3639 cm(-1) and are attributed to the inner surface hydroxyls, which are hydrogen bonded to the adsorbed water molecules. The hydroxyl stretching band of the inner hydroxyl is readily observed at 3621 cm(-1) in the Raman spectra of the CRTA treated formamide-intercalated kaolinites. The results of thermal analysis show that the amount of intercalated formamide between the kaolinite layers is independent of the presence of water. The Raman bands of the formamide in the CRTA-treated intercalated kaolinites are readily observed. Copyright 2001 Academic Press. PMID- 11397057 TI - The Effect of Temperature on Sorbitan Surfactant Monolayers. AB - The effect of temperature on four sorbitan ester (Span 20, sorbitan monolaurate; Span 40, sorbitan monopalmitate; Span 60, sorbitan monostearate; and Span 80, sorbitan monooleate) monolayers was analyzed. The covered temperature range was from 22 to 42 degrees C. Surface pressure-molecular area isotherms were measured with a Langmuir-type instrument. As the temperature was increased, the monolayers expanded more. This change can also be seen from the surface compressional modulus that was lowered as the temperature was increased. Also, the collapse pressure was lowered as the temperature was increased. Copyright 2001 Academic Press. PMID- 11397058 TI - Incorporation of Blood-Clotting Proteins into Phospholipid Langmuir Monolayers: A Fluorescence Microscopy Study. AB - Phospholipid monolayers adsorbed at an air-water interface are model cell membranes and have been used in this work to study interactions with blood clotting proteins. Factor I (non-membrane binding) was used as a control protein, and its association with L-alpha-dipalmitoylphosphatidylcholine Langmuir monolayers was compared to factor VII, a membrane-binding protein. Fluorescence micrographs indicated that factor I penetration of the lipid monolayers in the phase transition region occurred extensively, causing condensation of the lipid film. The association of factor I with phospholipid monolayers was deemed nonspecific. Factor VII was shown to associate with the periphery of lipid domains in the absence of calcium ions, causing flattening of domain edges. In the presence of calcium, factor VII induced expansion of the lipid monolayer. This effect is a specific interaction attributed to exposure of hydrophobic residues upon calcium binding, followed by protein association with lipid hydrocarbon chains. Copyright 2001 Academic Press. PMID- 11397059 TI - Langmuir Monolayers from Substituted 5'-Phenyl-m-terphenyl Carboxylic Acids. AB - A series of 5'-phenyl-m-terphenyl carboxylic acid (PTCA) derivatives with methyl, chloro, and fluoro substituents at both side phenyl rings have been synthesized and characterized as Langmuir monolayers at the air/water interface. A comparative analysis, based on the surface pressure and electric surface potential measurements under a variety of experimental conditions, is complemented with BAM images. Copyright 2001 Academic Press. PMID- 11397060 TI - Structural Modification Influences the Characteristics of Langmuir Monolayers from Aromatic Carboxylic Acids. AB - The molecular organization of purely aromatic, polyphenyl carboxylic acids, as Langmuir monolayers at the air/water interface, has been investigated by means of surface pressure and electric surface potential measurements upon film compression. The monolayer characteristics of the basic compound, a symmetrical triphenylbenzene (5'-phenyl-m-terphenyl) ring with a carboxylic group at the 4 position (namely 5'-phenyl-1,1' : 3',1"-terphenyl-4-carboxylic acid), are compared with those of its derivatives containing hydrophilic (nitro) or hydrophobic (phenyl) substituents. The nature of the substituent as well as its position (2' or 4') has a profound influence on the monolayer properties. The results are discussed in view of molecular orientation deduced from values of effective dipole moments. Copyright 2001 Academic Press. PMID- 11397061 TI - A PFG-NMR Study of Restricted Diffusion in Heterogeneous Polymer Particles. AB - The diffusion resistance to monomers during heterogeneous polymerization of polyolefin particles may have a significant effect on the observed activity. This diffusivity is, in general, unknown. To gain more information on this diffusion resistance in such systems, PFG-NMR has been used to measure the diffusion of organic solvents in various systems of porous polymer particles. In such systems the complex morphology and geometry demands careful analysis of the PFG-NMR attenuation curve. In this study, effects from restricted diffusion, domains having different diffusivity, and internal magnetic field gradients are expected. Thus, the obtained diffusivities have to be considered carefully, and a way to analyze the data taking these effects into account is presented. Copyright 2001 Academic Press. PMID- 11397062 TI - Mercury Porosimetry: Contact Angle Hysteresis of Materials with Controlled Pore Structure. AB - Mercury Porosimetry (MP) hysteresis is a commonly observed phenomenon in which mercury retention disguises further the overall hysteresis picture. This article introduces a new interpretation of the MP hysteresis based on the combined effect of pore structure networking and mercury contact angle variation occurring between the mercury penetration and retraction operations. To distinguish the contribution of each factor the following investigations were carried out. Nitrogen sorption (NP) and MP experiments were performed on samples of an anodic aluminum membrane and the results were interpreted in terms of the Corrugated Pore Structure Model (CPSM), i.e., CPSM-Nitrogen and CPSM-Mercury models, respectively. The simulation of the observed hysteresis data using the CPSM model enabled the evaluation of an identical for the two methods intrinsic pore size distribution (PSD) and cumulative surface area in perfect agreement with the respective BET value. Additionally, the CPSM analysis of data resulted in the evaluation of mercury contact angles, i.e., θ(p)=143 degrees and θ(r)=101.7 degrees for the MP penetration and retraction branches of the hysteresis loop, respectively. Moreover, CPSM-Mercury simulations of literature MP hysteresis data, valid for controlled-pore glasses and nuclepore membranes, led to the evaluation of contact angles, i.e., glasses: θ(p)=143 degrees, θ(r)=100.5-107.5 degrees and nuclepore: θ(p)=143 degrees, θ(r)=118- 121 degrees. The latter values are comparable with relevant literature data and approximate those determined for the anodic aluminum membrane. The CPSM model employed herein proved to be a flexible and reliable model for simulating the pertinent hysteresis loops by combining pore networking and contact angle hysteresis phenomena. Copyright 2001 Academic Press. PMID- 11397063 TI - Influence of Diffusion on the Kinetics of an Enzyme-Catalyzed Reaction in Gelatin Based Gels. AB - The influence of mass-transport limitations on the initial reaction rates of a lipase-catalyzed stereoselective esterification reaction has been investigated for two structurally different gelatin-based gels. The time to reach equilibrium is much longer in pelleted hydrogels (pseudo-solid aqueous gels; PAGs) than in pelleted microemulsion-based gels (MBGs). R/S-(+/-)-2-Octanol and hexanoic acid were used as substrates. The reaction takes place by imbibition of the substrate containing organic solvent into pores of the pelleted gel. To minimize the diffusion distances, the macroscopic surface areas of the gels were increased by granulating the gel pellets. The experimentally obtained initial reaction rates in granules were in good agreement with the theoretically obtained values from extrapolation to infinitely large areas. However, the still low initial reaction rates in the hydrogels compared to those in microemulsion-based gels cannot be explained by diffusion limitations. This finding was supported by the similar activation energies in both gels in granulated form. Changes in apparent molar standard enthalpy, entropy, and Gibbs energy for the activated complex formation were also estimated. The low reaction rate in hydrogels might thus be due to partial denaturation of the enzyme during the preparation step, to higher surface energy, or to the influence of a different solvent environment on the enzyme in these gels than in the microemulsion-based gels. Copyright 2001 Academic Press. PMID- 11397064 TI - Photodeposition of Prussian Blue Films on TiO(2): Additive Effect of Methanol and Influence of the TiO(2) Crystal Form. AB - The coupling of TiO(2) and transition metal complexes is attempted with the aim of higher functionalization of the TiO(2) photocatalyst. UV irradiation (lambda(ex)>300 nm) of a TiO(2) suspension containing equimolar aqueous solutions of FeCl(3) and K(3)[Fe(CN)(6)] forms uniform thin films of "water-insoluble Prussian blue" (PB, Fe(4)(3+) [Fe(II)(CN)(6)](3)) on the surface of TiO(2) particles. The PB photodeposition is enhanced significantly by the addition of a small amount of CH(3)OH in both the rutile and anatase TiO(2) systems. The activity of anatase TiO(2) is greater than that of rutile in the presence of CH(3)OH (2.46 M) by a factor of 1.6+/-0.2, whereas the activities are comparable in the absence of CH(3)OH. These results are discussed on the basis of a proposed reaction mechanism. Copyright 2001 Academic Press. PMID- 11397065 TI - The Structure and Properties of TiO(2)-Cu(II)-EDTA Ternary Surface Complexes. AB - The formation, composition, structure, and electrochemical properties of ternary surface complexes between copper(II) and ethylenediaminetetraacetate adsorbed on TiO(2) xerogels and on thin-film TiO(2) electrodes from solutions of varying pH have been studied by potentiometry, EPR spectroscopy, and electrochemical methods. The results strongly indicate that, in contrast to other organic ligands, B-type ternary surface complexes are formed in this system. The organic ligand forms an isolating layer between the surface of the TiO(2) electrode and the redox-active copper ions. Copyright 2001 Academic Press. PMID- 11397066 TI - The Influence of pH and Temperature on the Equilibrium and Dynamic Surface Tension of Aqueous Solutions of Sodium Oleate. AB - For aqueous solutions of sodium oleate, the dynamic surface tension (using the maximum bubble pressure technique) and the equilibrium surface tensions (using the du Nouy ring method) were studied. Experiments were carried out within the pH range 7 to 13 and at temperatures from 20 to 60 degrees C. From the equilibrium surface tension studies at 25 degrees C, a wide variation in surface activity was found to occur in the premicellar concentration range, depending on pH and oleate concentration, and this was explained on the basis of the formation of strongly surface-active premicellar species. It was also shown that the critical micelle concentration (cmc) of the system increased with pH within the pH range 7-12. Dynamic surface tension experiments were carried out from 20 to 60 degrees C at concentrations beyond the cmc in alkaline solutions, and the adsorption kinetics at the air/solution interface were analyzed using established theoretical diffusion models. From these data, the effective diffusion coefficients (D(eff)) for the oleate species were determined and the presence of an interfacial activation barrier in the diffusion process was confirmed. It was found that the D(eff) values obtained within the temperature range from 40 to 60 degrees C at 2 3 mM were in general agreement with previously reported values for the oleate dimer obtained by pulse-gradient FT-NMR. Furthermore, D(eff) significantly increased with temperature and also increased with concentration at higher temperatures (>40 degrees C). Copyright 2001 Academic Press. PMID- 11397067 TI - Effects of Nonionic Micelles on the Rate of Mononuclear Heterocyclic Rearrangement of (Z)-Phenylhydrazones of 5-Substituted 3-Benzoyl-1,2,4 oxadiazoles. AB - Nonionic Triton X-100 micelles solubilize the otherwise water-insoluble (Z) phenylhydrazones of some 5-substituted 3-benzoyl-1,2,4-oxadiazoles to an extent suitable for studying the occurrence of a general-base-catalyzed rearrangement in the presence of borate buffers (pH 9.6). The kinetic data, obtained at 40.0 degrees C over a wide range of surfactant concentrations, are found to conform to a reaction scheme which implies partitioning of the substrates and the base between water and the micellar pseudophase. Evidence that both the rate of the rearrangement reactions and the binding of the substrates to the micellar aggregates are primarily governed by the steric requirements of the 5-substituent group is obtained. Copyright 2001 Academic Press. PMID- 11397068 TI - Sucrose Derivative Surfactants Studied by Inverse Gas Chromatography. AB - Specific retention volumes of 12 solvents were measured at different temperatures on columns containing sucrose monopalmitate as their stationary phase. The influences of the temperature, adsorption effect in interface gas-liquid, and carrier gas flow on the chromatographic retention were analyzed. Henry's constants were calculated and the solubilities in the surfactant were obtained. The results were analyzed by the combination of Flory-Huggins and Hildebrand theories, and the solubility parameter of surfactant was obtained at different temperatures. The relation between solubility parameter and hydrophile-lipophile balance was discussed. Copyright 2001 Academic Press. PMID- 11397069 TI - Effects of Added Surfactant on the Dynamic Interfacial Tension Behavior of Acidic Oil/Alkaline Systems. AB - The effects of a ready-made surfactant (sodium dodecyl sulfate) on the dynamic interfacial tension between a model acidic oil (linoleic acid dissolved in paraffin oil) and various aqueous alkaline (NaOH) systems have been studied using pendant drop tensiometry at surfactant concentrations both below and above the critical micelle concentration (CMC). Below the CMC the added surfactant contributes significantly to a further reduction of interfacial tension of the reacting acid/alkaline system, whereas above the CMC the added surfactant plays an important role in damping the dynamic trends observed for the reactive system alone. Copyright 2001 Academic Press. PMID- 11397070 TI - Heterogemini Surfactants Based on Fatty Acid Synthesis and Interfacial Properties. AB - A series of novel nonionic surfactants based on fatty acids, each with two hydrophilic and two hydrophobic groups in the molecule (so-called Gemini surfactants), have been synthesized. The hydrophobic part of the surfactant, made from oleylnitrile, has a double bond in the middle of the chain to which the hydrophilic part is attached. One of the hydrophilic groups is a methyl-capped polyoxyethylene chain with 11 or 16 oxyethylene units, whereas the other is a secondary hydroxyl group. The reason for using the nitrile derivative of the fatty acid instead of a more conventional derivative such as an ester or amide is to achieve good hydrolytic stability, which is often demanded for cleaning applications. The cmc values of the surfactants were determined to be 0.2 and 0.4 mM for NIHG550 and NIHG750, respectively, using tensiometry and fluorescence. Pulsed-field gradient spin-echo NMR measurements gave similar but slightly higher values. The micelle size of NIHG750 was estimated to be on the order of 40 A, as calculated from the self-diffusion coefficient. The dynamic surface tension, gamma(t), was measured in the range 1 ms-1 s using a maximum bubble pressure instrument and analyzed in terms of asymptotic solutions to the Ward and Tordai equation. The results suggest that at the beginning the adsorption is essentially diffusion-controlled. However, close to equilibrium, the DST decays are not consistent with a diffusion-controlled adsorption mechanism. One important result from the present work is that the heterogeminis seem to align better than conventional surfactants at the air-liquid and solid-liquid interfaces. Copyright 2001 Academic Press. PMID- 11397071 TI - Concentrated CO(2)-in-Water Emulsions with Nonionic Polymeric Surfactants. AB - Concentrated CO(2)-in-water (C/W) emulsions are reported for amphiphiles containing alkylene oxide-, siloxane-, and fluorocarbon-based tails as a function of temperature and salinity. Poly(ethylene oxide)-b-poly(butylene oxide) (EO(15) b-BO(12)) can emulsify up to 70% CO(2) with droplet sizes from 2 to 4 &mgr;m in diameter, as determined by video-enhanced microscopy. This emulsion is stable over 48 h against both flocculation and coalescence. In contrast, it is extremely difficult to form concentrated water-in-CO(2) (W/C) emulsions with surfactants containing alkylene oxide moieties due to limited solvation of such tails by CO(2). In several cases, C/W emulsions are formed even when the surfactant prefers CO(2). This violation of Bancroft's rule may be attributed in part to the low viscosity of the compressed CO(2), which governs several mass and momentum transport mechanisms relevant to emulsion formation and stabilization. For the first time, W/C microemulsions are observed in a system with a nonionic amphiphile, namely F(CF(2)CF(2))(3-8)CH(2)CH(2)O(CH(2)CH(2)O)(10-15)H. For the same system, the emulsion morphology changes from C/W to W/C as the temperature increases. The electrical conductivity of C/W emulsions is predicted successfully as a function of the dispersed phase volume fraction of CO(2) with Maxwell's theory for inhomogeneous systems. Copyright 2001 Academic Press. PMID- 11397072 TI - Thermodynamic Study on Vesicle Formation and Adsorption of Decyltrimethylammonium Decyl Sulfate. AB - The surface tension of an aqueous solution of decyltrimethylammonium decyl sulfate (DeTADeS) was measured as a function of temperature T at various molalities &mcirc; under atmospheric pressure. DeTADeS has been found to form equilibrium multilamellar vesicles (MLV) spontaneously. The surface density, the entropies of adsorption, and the entropy of vesicle formation are evaluated. The mechanism of formation of equilibrium vesicles is investigated from the standpoint of thermodynamics and from the comparison of the results with those of the micelle-forming systems. From the relatively small change of the surface density Gamma;(H) on T at a given &mcirc;, the adsorbed film is implied to be tightly packed due to the strong electrostatic attraction between the polar headgroups. The energy change associated with adsorption from the vesicular state per mole of surfactant Delta(V)(H)u is positive in the entire temperature range; thus, the curved bilayer in MLV is energetically more favorable than the planar adsorbed film. From the negative values of the entropy of vesicle formation Delta(W)(V)s, it is concluded that vesicle formation is driven by enthalpy whereas micelle formation is mostly entropy driven. Copyright 2001 Academic Press. PMID- 11397073 TI - Transport Properties of Aqueous Solutions of Alkyltrimethylammonium Bromide Surfactants at 25 degrees C. AB - Intradiffusion coefficients, D, of n-alkyltrimethylammonium bromides [CH(3) (CH(2))(n-1)-N(CH(3))(3)Br, C(n)TAB] (n=6, 8, 10, 12) in mixtures with heavy water were measured by the PGSE-NMR technique at 25 degrees C. The experimental data permitted evaluation of the influence of the alkyl chain length on the surfactant self-aggregation process. For all the surfactants considered, the D trend showed a slope change corresponding to the critical micellar composition (cmc). In the premicellar composition range, D decreased linearly with the square root of the surfactant molality. The D values extrapolated at infinite dilution were related to the limiting mutual diffusion coefficients, determined through the Taylor dispersion technique. In the micellar composition range, solubilized tetramethylsilane (TMS) molecules were used to determine the micelle intradiffusion coefficient, D(M), from which the aggregate radii and the aggregation numbers were obtained. The decreasing trend of D(M) with increasing surfactant molality was interpreted in terms of interparticle electrostatic repulsion. D(M) values allowed evaluation of the Gouy-Chapman layer thickness. The solvent intradiffusion coefficient in the heavy water-C(n)TAB mixtures, D(w), was also measured. It decreased with increasing surfactant molality. For n=8, 10, 12 the D(w) trend presented a slope change at the cmc, which could be ascribed to the strong decrease in hydration of surfactant molecules upon micellization. Because of its short hydrophobic tail, C(6)TAB exhibited peculiar aggregation behavior. Its cmc, which is poorly marked, is lower than the value predicted by extrapolating the cmc values obtained for the other terms of the series. The C(6)TAB aggregates do not solubilize TMS molecules; the estimated aggregation number is extremely low ( approximately 3). Finally, no abrupt slope change in the solvent intradiffusion coefficient trend was detected. This evidence suggests that C(6)TAB molecules do not micellize in aqueous solution, but form trimers in which the surfactant hydrophobic tails are not hidden from contact with water molecules. Copyright 2001 Academic Press. PMID- 11397074 TI - Thin-Film Morphologies of Random Copolymers as Functions of Thermal History. AB - Series polymers of butyl methacrylate with various contents of 3 (trimethoxysilyl)propyl methacrylate (MSMA) that introduces crosslinking networks among the macromolecules upon hydrolysis and self-condensation have been synthesized by free radical polymerization, and the influence of crosslinking density on the film properties has been examined. The polymer solutions were spin cast over a layer of polystyrene brush to yield homogeneous polymer films. When the films of about 30 nm thick were exposed to moisture and then heated at 60 degrees C for hydrolysis and self-condensation of MSMA groups, the initially flat surfaces became slightly coarser but without apparent dewetting. Further annealing at 140 degrees C resulted in dewetting of the thin films, whose morphologies in thermodynamic equilibrium were related to the chemical compositions of the polymers. The polymers having higher contents of MSMA exhibited significantly reduced dewetting at the high temperature, due to the higher density of crosslinking networks that restricted the molecular mobility. In contrast to the thin films of about 30 nm thick, thicker films (about 100 nm) showed only a slight dewetting, even non-dewetting at the elevated temperature. Copyright 2001 Academic Press. PMID- 11397075 TI - Nonsingular van der Waals Potential and Its Contributions to Gravitational Coagulation. AB - The coagulation and the stability of dilute colloids at high Peclet number were studied in consideration of the universal nonsingular van der Waals interactions recently developed by J. X. Lu and W. H. Marlow (1995, Phys. Rev. Lett. 74, 1724). The capture efficiency of gravitational coagulation of uncharged colloids was found to be diminished due to the effect of finite molecular size (EFMS). The gravitational coagulation stability diagram of charged colloids was also found to be shifted when the gravitational convection was much stronger than the Brownian diffusion. Copyright 2001 Academic Press. PMID- 11397076 TI - Constraints on Contact Angles for Multiple Phases in Thermodynamic Equilibrium. AB - For three or more fluid phases in thermodynamic equilibrium and in contact with a solid surface, the Young equation can be used to find relations between the contact angles for different pairs of fluids. For an n-fluid-phase system, n(n 1)/2 contact angles can be defined, but there are (n-1)(n-2)/2 constraints between them, leaving only n-1 independent values of the contact angle. These constraints are very powerful in limiting and determining possible types of wetting behavior. The consequences are discussed for three- and four-phase flow. They have important applications for the understanding of gas injection processes in petroleum reservoirs. Copyright 2001 Academic Press. PMID- 11397077 TI - Does a stretched DNA structure dictate the helical geometry of RecA-like filaments? AB - Proteins in the RecA/Rad51/RadA/UvsX family form helical filaments on DNA in which the DNA is stretched and untwisted. A comparison of the average helical parameters of these filaments from five different proteins, obtained from archaea, eubacteria and eukaryotes, suggests that an intrinsic state of DNA may be responsible for the conservation of these particular filament forms across evolution. In this view, these proteins stabilize this existing state of DNA, rather than induce a novel conformation. PMID- 11397078 TI - Protein chemistry at membrane interfaces: non-additivity of electrostatic and hydrophobic interactions. AB - Non-specific binding of proteins and peptides to charged membrane interfaces depends upon the combined contributions of hydrophobic (DeltaG(HPhi)) and electrostatic (DeltaG(ES)) free energies. If these are simply additive, then the observed free energy of binding (DeltaG(obs)) will be given by DeltaG(obs)=DeltaG(HPhi)+DeltaG(ES), where DeltaG(HPhi)=-sigma(NP)A(NP) and DeltaG(ES)=zFphi. In these expressions, A(NP) is the non-polar accessible area, sigma(NP) the non-polar solvation parameter, z the formal peptide valence, F the Faraday constant, and phi the membrane surface potential. But several lines of evidence suggest that hydrophobic and electrostatic binding free energies of proteins at membrane interfaces, such as those associated with cell signaling, are not simply additive. In order to explore this issue systematically, we have determined the interfacial partitioning free energies of variants of indolicidin, a cationic proline-rich antimicrobial peptide. The synthesized variants of the 13 residue peptide covered a wide range of hydrophobic free energies, which allowed us to examine the effect of hydrophobicity on electrostatic binding to membranes formed from mixtures of neutral and anionic lipids. Although DeltaG(obs) was always a linear function of DeltaG(HPhi), the slope depended upon anionic lipid content: the slope was 1.0 for pure, zwitterionic phosphocholine bilayers and 0.3 for pure phosphoglycerol membranes. DeltaG(obs) also varied linearly with surface potential, but the slope was smaller than the expected value, zF. As observed by others, this suggests an effective peptide valence z(eff) that is smaller than the formal valence z. Because of our systematic approach, we were able to establish a useful rule-of-thumb: z(eff) is reduced relative to z by about 20 % for each 3 kcal mol(-1) (1 kcal=4.184 kJ) favorable increase in DeltaG(HPhi). For neutral phosphocholine interfaces, we found that DeltaG(obs) could be predicted with remarkable accuracy using the Wimley-White experiment-based interfacial hydrophobicity scale. PMID- 11397079 TI - Observation of large, non-covalent globin subassemblies in the approximately 3600 kDa hexagonal bilayer hemoglobins by electrospray ionization time-of-flight mass spectrometry. AB - A non-covalent globin subassembly comprising 12 globin chains (204 to 214 kDa) was observed directly by electrospray ionization time-of-flight mass spectrometry in the native hexagonal bilayer hemoglobins from the oligochaetes Lumbricus terrestris and Tubifex tubifex, the polychaetes Tylorrhynchus heterochaetus, Arenicola marina, Amphitrite ornata and Alvinella pompejana, the leeches Macrobdella decora, Haemopis grandis and Nephelopsis oscura and the chlorocruorin from the polychaete Myxicola infundibulum, over the pH range 3.5-7.0. The Hb from the deep-sea polychaete Alvinella exhibited in addition, peaks at approximately 107 kDa and at approximately 285 kDa, which were assigned to subassemblies of six globin chains and of 12 globin chains with three non-globin linker chains, respectively. The experimental masses decreased slightly with increased de clustering potential (60 to 160 V) and were generally 0.1 to 0.2 % higher than the calculated masses, due probably to complexation with cations and water molecules. PMID- 11397080 TI - Domain 1.1 of the sigma(70) subunit of Escherichia coli RNA polymerase modulates the formation of stable polymerase/promoter complexes. AB - The sigma 70 (sigma(70)) subunit of Escherichia coli RNA polymerase specifies transcription from promoters that are responsible for basal gene expression during vegetative growth. When sigma(70) is present within polymerase holoenzyme, two of its domains, 2.4 and 4.2, interact with sequences within the -10 and -35 regions, respectively, of promoter DNA. However, in free sigma(70), DNA binding is prevented by domain 1.1, the N-terminal domain of the protein. Previous work has demonstrated that the presence of domain 1.1 is required for efficient transcription initiation at the lambda promoter P(R). To investigate whether this is a general property of domain 1.1, we have used five promoters to compare polymerases with and without domain 1.1 in in vitro transcription assays, and in assays assessing the formation and decay of stable, pretranscription complexes. We find that the absence of domain 1.1 does not render the polymerase defective at all of these promoters. Depending on the promoter, the absence of domain 1.1 can promote or inhibit transcription initiation by affecting the formation of stable pretranscription complexes. However, domain 1.1 does not affect the stability of these complexes once they are formed. For polymerases containing domain 1.1, the efficiency of stable complex formation correlates with how well the -10 and -35 regions of a promoter match the ideal sigma(70) recognition sequences. However, when domain 1.1 is absent, having this match becomes less important in determining how efficiently stable complexes are made. We suggest that domain 1.1 influences initiation by constraining polymerase to assess a promoter primarily by the fitness of its -10 and -35 regions to the canonical sequences. PMID- 11397081 TI - Repression of transcription initiation at 434 P(R) by 434 repressor: effects on transition of a closed to an open promoter complex. AB - The lambdoid bacteriophage repressors function both as transcription activators and repressors. Regulation of transcription at the adjacent, but divergent promoters, P(RM) and P(R), determines the phage's choice between the lytic and lysogenic development pathways. Here, we demonstrate that 434 repressor bound at 434 O(R)1 alone is not sufficient to repress transcription from 434 P(R,) but that 434 repressor bound at 434 O(R)2 alone is necessary and sufficient to repress P(R )transcription. This is different from what occurs in the related bacteriophage lambda, in which binding of lambda repressor to either lambdaO(R)1 or lambdaO(R)2 represses transcription from lambdaP(R). The combined results of gel mobility shift and KMnO(4) footprinting assays show that while 434 repressor binding to 434 O(R)2 does not preclude RNA polymerase binding at the P(R) promoter, it does prevent it from forming open complexes at this promoter. The RNA polymerase-P(R) complexes that form in the presence of repressor are heparin resistant and the DNA is not melted. This observation indicates that 434 repressor bound at 434 O(R)2 inhibits transcription initiation at the P(R) promoter by "locking" the RNA polymerase-P(R) complex into an inactive state instead of "blocking" the access of RNA polymerase to promoter DNA. PMID- 11397082 TI - The basal transcription factors TBP and TFB from the mesophilic archaeon Methanosarcina mazeii: structure and conformational changes upon interaction with stress-gene promoters. AB - Transcription of archaeal non-stress genes involves the basal factors TBP and TFB, homologs of the eucaryal TATA-binding protein and transcription factor IIB, respectively. No comparable information exists for the archaeal molecular chaperone, stress genes hsp70(dnaK), hsp40(dnaJ), and grpE. These do not occur in some archaeal species, but are present in others possibly due to lateral transfer from bacteria, which provides a unique opportunity to study regulation of stress inducible bacterial genes in organisms with eukaryotic-like transcription machinery. Among the Archaea with the genes, those from the mesophilic methanogen Methanosarcina mazeii are the only ones whose basal (constitutive) and stress induced transcription patterns have been determined. To continue this work, tbp and tfb were cloned from M. mazeii, sequenced, and the encoded recombinant proteins characterized in solution, separately and in complex with each other and with DNA. M. mazeii TBP ranks among the shortest within Archaea and, contrary to other archaeal TBPs, it lacks tryptophan or an acidic tail at the C terminus and has a basic N-terminal third. M. mazeii TFB is similar in length to archaeal and eucaryal homologs and all have a zinc finger and HTH motifs. Phylogenetically, the archaeal and eucaryal proteins form separate clusters and the M. mazeii molecules are closer to the homologs from Archaeoglobus fulgidus than to any other. Antigenically, M. mazeii TBP and TFB are close to archaeal homologs within each factor family, but the two families are unrelated. The purified recombinant factors were functionally active in a cell-free in vitro transcription system, and were interchangeable with the homologs from Methanococcus thermolithotrophicus. The M. mazeii factors have a similar secondary structure by circular dichroism (CD). The CD spectra changed upon binding to the promoters of the stress genes grpE, dnaK, and dnaJ, with the changes being distinctive for each promoter; in contrast, no effect was produced by the promoter of a non stress-gene. Factor(s)-DNA modeling predicted that modifications of H bonds are caused by TBP binding, and that these modifications are distinctive for each promoter. It also showed which amino acid residues would contact an extended TATA box with a B recognition element, and evolutionary conservation of the TBP-TFB DNA complex orientation between two archaeal organisms with widely different optimal temperature for growth (37 and 100 degrees C). PMID- 11397083 TI - Four-way junctions in antisense RNA-mRNA complexes involved in plasmid replication control: a common theme? AB - In several groups of bacterial plasmids, antisense RNAs regulate copy number through inhibition of replication initiator protein synthesis. In plasmid R1, we have recently shown that the inhibitory complex between the antisense RNA (CopA) and its target mRNA (CopT) is characterized by the formation of two intermolecular helices, resulting in a four-way junction structure and a side-by side helical alignment. Based on lead-induced cleavage and ribonuclease (RNase) V(1) probing combined with molecular modeling, a strikingly similar topology is supported for the complex formed between the antisense RNA (Inc) and mRNA (RepZ) of plasmid Col1b-P9. In particular, the position of the four-way junction and the location of divalent ion-binding site(s) indicate that the structural features of these two complexes are essentially the same in spite of sequence differences. Comparisons of several target and antisense RNAs in other plasmids further indicate that similar binding pathways are used to form the inhibitory antisense target RNA complexes. Thus, in all these systems, the structural features of both antisense and target RNAs determine the topologically possible and kinetically favored pathway that is essential for efficient in vivo control. PMID- 11397084 TI - Sequence-specific recognition of DNA in the nucleosome by pyrrole-imidazole polyamides. AB - The ability of DNA-binding proteins to recognize their cognate sites in chromatin is restricted by the structure and dynamics of nucleosomal DNA, and by the translational and rotational positioning of the histone octamer. Here, we use six different pyrrole-imidazole polyamides as sequence-specific molecular probes for DNA accessibility in nucleosomes. We show that sites on nucleosomal DNA facing away from the histone octamer, or even partially facing the histone octamer, are fully accessible and that nucleosomes remain fully folded upon ligand binding. Polyamides only failed to bind where sites are completely blocked by interactions with the histone octamer. Removal of the amino-terminal tails of either histone H3 or histone H4 allowed these polyamides to bind. These results demonstrate that much of the DNA in the nucleosome is freely accessible for molecular recognition in the minor groove, and also support a role for the amino-terminal tails of H3 and H4 in modulating accessibility of nucleosomal DNA. PMID- 11397085 TI - Subcellular localization and oligomerization of the Arabidopsis thaliana somatic embryogenesis receptor kinase 1 protein. AB - The Arabidopsis thaliana somatic embryogenesis receptor kinase 1 (AtSERK1) gene is expressed in developing ovules and early embryos. AtSERK1 is also transiently expressed during somatic embryogenesis. The predicted AtSERK1 protein contains an extracellular domain with a leucine zipper motif followed by five leucine-rich repeats, a proline-rich region, a single transmembrane region and an intracellular kinase domain. The AtSERK1 cDNA was fused to two different variants of green fluorescent protein (GFP), a yellow-emitting GFP (YFP) and a cyan emitting GFP (CFP), and transiently expressed in both plant protoplasts and insect cells. Using confocal laser scanning microscopy it was determined that the AtSERK1-YFP fusion protein is targeted to plasma membranes in both plant and animal cells. The extracellular leucine-rich repeats, and in particular the N linked oligosaccharides that are present on them appear to be essential for correct localization of the AtSERK1-YFP protein. The potential for dimerization of the AtSERK1 protein was investigated by measuring the YFP/CFP fluorescence emission ratio using fluorescence spectral imaging microscopy. This ratio will increase due to fluorescence resonance energy transfer if the AtSERK1-CFP and AtSERK1-YFP fusion proteins interact. In 15 % of the cells the YFP/CFP emission ratio for plasma membrane localized AtSERK1 proteins was enhanced. Yeast-protein interaction experiments confirmed the possibility for AtSERK1 homodimerization. Elimination of the extracellular leucine zipper domain reduced the YFP/CFP emission ratio to control levels indicating that without the leucine zipper domain AtSERK1 is monomeric. PMID- 11397086 TI - Kinetics of the AHL regulatory system in a model biofilm system: how many bacteria constitute a "quorum"? AB - Acylated homoserine lactones (AHLs) regulate a wide variety of phenotypes in Gram negative bacteria. Most research suggests that AHL-mediated phenotypes are not expressed in populations until late logarithmic phase or stationary phase. Here, we model how the concentration of AHLs inside bacterial cells and in a biofilm changes over time as a function of population growth rate, diffusion of AHLs and the rate of autoinduction. Our theoretical results show that the concentration of AHLs inside a single bacterium (and by implication induction of a phenotype) has a non-trivial behaviour over time, and often exhibits a rapid increase early in population growth. This rapid increase is followed by a plateau, followed by another rise in the concentration of AHLs, to a second plateau. High concentrations of AHLs inside the bacterial cell early in population growth are positively affected by slow diffusion rates out of the cell and the biofilm, slow bacterial growth rates and fast autoinduction. In contrast, fast growth rates, slow autoinduction rates and high diffusion rates result in a high concentration plateau in stationary phase. More generally, the density-dependent nature of AHL regulation can be viewed as a trade-off between factors that dilute intracellular concentrations of AHLs (diffusion out of the cell, cell division), and those that increase concentrations (a slowing or restriction of diffusion or growth, or autoinduction). These results suggest that expression of AHL-mediated phenotypes can occur at relatively low cell densities and low external/environmental AHL concentrations. PMID- 11397087 TI - Yet another numbering scheme for immunoglobulin variable domains: an automatic modeling and analysis tool. AB - A common residue numbering scheme for all immunoglobulin variable domains (immunoglobulin light chain lambda (V(lambda)) and kappa (V(kappa)) variable domains, heavy chain variable domains (V(H)) and T-cell receptor alpha (V(alpha)), beta (V(beta)), gamma (V(gamma)) and delta (V(delta)) variable domains) has been devised. Based on the spatial alignment of known three dimensional structures of immunoglobulin domains, it places the alignment gaps in a way that minimizes the average deviation from the averaged structure of the aligned domains. This residue numbering scheme was applied to the immunoglobulin variable domain structures in the PDB database to automate the extraction of information on structural variations in homologous positions of the different molecules. A number of methods are presented that allow the automated projection of information derived from individual structures or from the comparison of multi structure alignments onto a graphical representation of the sequence alignment. PMID- 11397088 TI - Selection, characterization and x-ray structure of anti-ampicillin single-chain Fv fragments from phage-displayed murine antibody libraries. AB - Single-chain Fv (scFv) antibody libraries were constructed from mice immunized with an ampicillin-bovine serum albumin conjugate. Several antibodies with specificity for intact ampicillin were selected by phage display and characterized. The antibody scFv fragment aL2 binds to intact ampicillin and shows no detectable cross-reactivity with hydrolyzed ampicillin. We determined the X-ray structures of two crystal forms of w.t. aL2, which differ mainly in the side-chain conformation of Trp H109 (according to a new consensus nomenclature Kabat residue number H95) in the extremely short (three residues) CDR H3 and the presence or absence of a well-resolved molecule of 2-methyl-pentane-2,4-diol in the bottom of the binding pocket. Attempts to co-crystallize aL2 with its antigen or to diffuse ampicillin into the wild-type aL2 crystals were unsuccessful, since crystal contacts obstruct the binding pocket. However, a mutant with two point mutations near the N terminus (Gln H6 replaced by Glu and Ala H10 (Kabat H9) replaced by Gly) crystallized in a form compatible with antigen-binding. Although the mutations affect the conformation of framework I, the conformations of the binding pocket of the uncomplexed wild-type aL2 and of the mutant complex were almost identical. The structure explains the specificity of the antibody for intact ampicillin and the degree of cross-reactivity of aL2 with a wide variety of ampicillin analogs. This antibody system will be very useful as a diagnostic reagent for antibiotics use and abuse, as a model for the effect of expression of antibiotic binding molecules in Escherichia coli, and for directed evolution towards high antibiotic resistance. PMID- 11397089 TI - The influence of the buried glutamine or glutamate residue in position 6 on the structure of immunoglobulin variable domains. AB - Immunoglobulin V(H) domain frameworks can be grouped into four distinct types, depending on the main-chain conformation of framework 1. Based on the analysis of over 200 X-ray structures representing more than 100 non-redundant V(H) domain sequences, we have come to the conclusion that the marked structural variability of the V(H) framework 1 region is caused by three residues: the buried side-chain of H6, which can be either a glutamate or a glutamine residue, the residue in position H7, which may be proline only if H6 is glutamine, and by H9 (H10 according to a new consensus nomenclature), which has to be either glycine or proline if H6 is a glutamate residue. In natural antibodies, these three residues are encoded in combinations that are compatible with each other and with the rest of the structure and therefore will yield functional molecules. However, the degenerate primer mixtures commonly used for PCR cloning of antibody fragments can and frequently do introduce out-of-context mutations to combinations that can lead to severe reduction of stability, production yield and antigen affinity. PMID- 11397090 TI - The importance of framework residues H6, H7 and H10 in antibody heavy chains: experimental evidence for a new structural subclassification of antibody V(H) domains. AB - The N-terminal segment (FR-H1) of the heavy chain (V(H)) of antibodies shows significant conformational variability correlating with the nature of the amino acids H6, H7 and H10 (Kabat H9). In this study, we have established a causal relationship between the local sequence and the structure of this framework region and linked this relationship to important biophysical properties such as affinity, folding yield and stability. We have generated six mutants of the scFv fragment aL2, covering some of the most abundant amino acid combinations in positions H6, H7 and H10 (according to a new consensus nomenclature, Kabat H9). For the aL2 wild-type (w.t.) with the sequence 6(Q)7(P)10(A) and for two of the mutants, the X-ray structures have been determined. The structure of the triple mutant aL2-6(E)7(S)10(G) shows the FR-H1 backbone conformations predicted for this amino acid combination, which is distinctly different from the structure of the w.t, thus supporting our hypothesis that these residues determine the conformation of this segment. The mutant aL2-6(E)7(P)10(G) represents a residue combination not occurring in natural antibody sequences. It shows a completely different, unique structure in the first beta-strand of V(H), not observed in natural Fv fragments and forms a novel type of diabody. Two V(H) domains of the mutant associate by swapping the first beta-strand. Concentration-dependent changes in Trp fluorescence indicate that this dimerization also occurs in solution. The mutations in amino acids H6, H7 and H10 (Kabat H9) influence the dimerization behavior of the scFv and its thermodynamic stability. All the observations reported here have practical implications for the cloning of Fv fragments with degenerate primers, as well as for the design of new antibodies by CDR grafting or synthetic libraries. PMID- 11397091 TI - In-vitro selection of highly stabilized protein variants with optimized surface. AB - Thermostable proteins are of prime importance in protein science, but it has remained difficult to develop general strategies for stabilizing a protein. Site directed mutagenesis based on comparisons with thermophilic homologs is rarely successful because the sequence differences are too numerous and dominated by neutral mutations. Here we used a method of directed evolution to increase the stability of a mesophilic protein, the cold shock protein Bs-CspB from Bacillus subtilis. It differs from its thermophilic counterpart Bc-Csp from Bacillus caldolyticus at 12 surface-exposed positions. To elucidate the stabilizing potential of exposed amino acid residues, six of these variant positions were randomized by saturation mutagenesis, the corresponding library of sequences was inserted into the gene-3-protein of the filamentous phage fd, and stabilized variants were selected by the Proside technique. Proside links the increased protease resistance of stabilized protein variants with the infectivity of the phage. Many strongly stabilized variants of Bs-CspB were identified in two selections, one in the presence of a denaturant and the other at elevated temperature. Several of them are significantly more stable than the naturally thermostable homolog Bc-Csp, and the best variant reaches Tm-Csp (the homolog from the hyperthermophile Thermotoga maritima) in stability. Remarkably, this variant differs from Tm-Csp at five and from Bc-Csp at all six randomized positions. This indicates that proteins can be strongly stabilized by many different sets of surface mutations, and Proside selects them efficiently from large libraries. The course of the selection could be directed by the conditions. In an ionic denaturant non-polar surface interactions were optimized, whereas at elevated temperature variants with improved electrostatics were selected, pointing to two different strategies for stabilization at protein surfaces. PMID- 11397092 TI - Structural studies of the tRNA domain of tmRNA. AB - tmRNA is a small, stable prokaryotic RNA. It rescues ribosomes that have become stalled during the translation of mRNA fragments lacking stop codons, or during periods of tRNA scarcity. It derives its name from the presence of two separate domains, one that functions as a tRNA, and another that serves as an mRNA. We have carried out modeling and transient electric birefringence studies to determine the angle between the acceptor stem and anticodon stem of the tRNA domain of Eschericia coli tmRNA. The results of the modeling studies yielded an interstem angle of 110 degrees, in agreement with the lower end of the range of angles (111 degrees -137 degrees ) determined experimentally for various solution conditions. The range of experimental angles is greater than the angles observed for any of the tRNA crystal structures, in line with the presence of a shortened D stem. The secondary structure of the tRNA domain is conserved for all known tmRNA sequences, so we propose that the angle is also conserved. These results also suggest that the region of tmRNA between P2a and P2b may interact with the decoding site of the ribosome. PMID- 11397093 TI - Molecular basis for immune complex recognition: a comparison of Fc-receptor structures. AB - Once antigen is opsonised by IgG it is removed from the circulation by Fcgamma receptor expressing cells. Fcgamma-receptors are type I transmembrane molecules that carry extracellular parts consisting of two or three immunoglobulin domains. Previously solved structures of Fc-receptors reveal that the N-terminal two Ig like domains are arranged in a steep angle forming a heart-shaped structure. The crystal structure of the FcgammaRIII/hIgG1-Fc-fragment demonstrated that the Fc fragment is recognised through loops of the C-terminal receptor domain of the FcgammaRIII. As the overall structure of the FcRs and their Ig ligands are very similar we modelled the Ig complexes with FcgammaRI, FcgammaRII and FcepsilonRIalpha based on the FcgammaRIII/hIgG1-Fc-fragment structure. The obtained models are consistent with the observed biochemical data and may explain the observed specificity and affinities. PMID- 11397094 TI - Camel lactoferrin, a transferrin-cum-lactoferrin: crystal structure of camel apolactoferrin at 2.6 A resolution and structural basis of its dual role. AB - Camel lactoferrin is the first protein from the transferrin superfamily that has been found to display the characteristic functions of iron binding and release of lactoferrin as well as transferrin simultaneously. It was remarkable to observe a wide pH demarcation in the release of iron from two lobes. It loses 50 % iron at pH 6.5 and the remaining 50 % iron is released only at pH values between 4.0 and 2.0. Furthermore, proteolytically generated N and C-lobes of camel lactoferrin showed that the C-lobe lost iron at pH 6.5, while the N-lobe lost it only at pH less than 4.0. In order to establish the structural basis of this striking observation, the purified camel apolactoferrin was crystallized. The crystals belong to monoclinic space group C2 with unit cell dimensions a=175.8 A, b=80.9 A, c=56.4 A, beta=92.4 degrees and Z=4. The structure has been determined by the molecular replacement method and refined to an R-factor of 0.198 (R-free=0.268) using all the data in the resolution range of 20.0-2.6 A. The overall structure of camel apolactoferrin folds into two lobes which contain four distinct domains. Both lobes adopt open conformations indicating wide distances between the iron binding residues in the native iron-free form of lactoferrin. The dispositions of various residues of the iron binding pocket of the N-lobe of camel apolactoferrin are similar to those of the N-lobe in human apolactoferrin, while the corresponding residues in the C-lobe show a striking similarity with those in the C-lobes of duck and hen apo-ovotransferrins. These observations indicate that the N-lobe of camel apolactoferrin is structurally very similar to the N-lobe of human apolactoferrin and the structure of the C-lobe of camel apolactoferrin matches closely with those of the hen and duck apo-ovotransferrins. These observations suggest that the iron binding and releasing behaviour of the N-lobe of camel lactoferrin is similar to that of the N-lobe of human lactoferrin, whereas that of the C-lobe resembles those of the C-lobes of duck and hen apo ovotransferrins. Hence, it correlates with the observation of the N-lobe of camel lactoferrin losing iron at a low pH (4.0-2.0) as in other lactoferrins. On the other hand, the C-lobe of camel lactoferrin loses iron at higher pH (7.0-6.0) like transferrins suggesting its functional similarity to that of transferrins. Thus, camel lactoferrin can be termed as half lactoferrin and half transferrin. PMID- 11397095 TI - Recognition of pre-formed and flexible elements of an RNA stem-loop by nucleolin. AB - Nucleolin is an abundant nucleolar protein which is essential for ribosome biogenesis. The first two of its four tandem RNA-binding domains (RBD12) specifically recognize a stem-loop structure containing a conserved UCCCGA sequence in the loop called the nucleolin-recognition element (NRE). We have determined the structure of the consensus SELEX NRE (sNRE) by NMR spectroscopy. In both the free and bound RNA the top part of the stem forms a loop E (or S turn) motif. In the absence of protein, the structure of the hairpin loop is not well defined due to conformational heterogeneity, and appears to be in equilibrium between two families of conformations. Titrations of RBD1, RBD2, and RBD12 with the sNRE show that specific binding requires RBD12. In complex with RBD12, the hairpin loop interacts specifically with the protein and adopts a well defined structure which shares some of the features of the free form. The loop E motif also has specific interactions with the protein. Implications of these findings for the mechanism of recognition of RNA structures by modular proteins are discussed. PMID- 11397096 TI - The B(12)-binding subunit of glutamate mutase from Clostridium tetanomorphum traps the nucleotide moiety of coenzyme B(12). AB - Glutamate mutase from Clostridium tetanomorphum binds coenzyme B(12) in a base off/His-on form, in which the nitrogenous ligand of the B(12)-nucleotide function is displaced from cobalt by a conserved histidine. The effect of binding the B(12)-nucleotide moiety to MutS, the B(12)-binding subunit of glutamate mutase, was investigated using NMR spectroscopic methods. Binding of the B(12)-nucleotide to MutS was determined to occur with K(d)=5.6(+/-0.7) mM and to be accompanied by a specific conformational change in the protein. The nucleotide binding cleft of the apo-protein, which is formed by a dynamic segment with propensity for partial alpha-helical conformation (the "nascent" alpha-helix), becomes completely structured upon binding of the B(12)-nucleotide, with formation of helix alpha1. In contrast, the segment containing the conserved residues of the B(12)-binding Asp-x-His-x-x-Gly motif remains highly dynamic in the protein/B(12)-nucleotide complex. From relaxation studies, the time constant tau, which characterizes the time scale for the formation of helix alpha1, was estimated to be about 30 micros (15)N and was the same in both, apo-protein and nucleotide-bound protein. Thus, the binding of the B(12)-nucleotide moiety does not significantly alter the kinetics of helix formation, but only shifts the equilibrium towards the structured fold. These results indicate MutS to be structured in such a way, as to be able to trap the nucleotide segment of the base-off form of coenzyme B(12) and provide, accordingly, the first structural clues as to how the process of B(12)-binding occurs. PMID- 11397097 TI - Surface map comparison: studying function diversity of homologous proteins. AB - A simplified protein surface cartography approach has been developed to assist in the analysis of surface features in homologous families, and thus to predict conservation or divergence of protein functions and protein-protein interaction patterns. A spherical approximation of protein surface was used, with a focus on charged and hydrophobic residues. The resulting surface map allows for qualitative analysis and comparison of surfaces of proteins, but can also be used to define a simple numerical measure of map similarity between two or more proteins. The latter was shown to be useful for function based classifications within large protein families. Surface map analysis was tested on several test cases: haemoglobins, death domains and TRAF domains. It was shown that surface map comparison allows a better function prediction than general sequence analysis methods and can reproduce known examples of functional variation within a divergent group of proteins. In another example, we predict novel, unexpected sets of common functional properties for seemingly distant members of a large group of divergent proteins. The method was also shown to be robust enough to allow using protein models from comparative modelling instead of experimental structures. PMID- 11397098 TI - Osteogenesis imperfecta murine: interaction between type I collagen homotrimers. AB - Types I, II, and III collagens are believed to have evolved from the same homotrimer ancestor and they have substantial sequence homology, but type I molecules are alpha1(I)(2)alpha2(I) heterotrimers, unlike homotrimeric types II and III. It is believed that the alpha2(I) chain first appeared in lower vertebrates and that it plays a particularly important role in bone formation. For instance, spontaneous mutations resulting in non- functional alpha2 chains and formation of type I homotrimers cause severe bone pathology (osteogenesis imperfecta) in humans and in animals. However, the exact role of the alpha2 chain is not known. Here, we report measurements of intermolecular forces between collagen helices in native and reconstituted fibers composed of type I homotrimers, heterotrimers and their mix. For comparison, we report forces between type II homotrimers in reconstituted fibers. In agreement with previous studies, we find that the absence of the alpha2 chain reduces temperature-favored attraction between collagen helices, either because of the difference in amino acid sequence of the alpha1 and alpha2 chains or because of more extensive post translational modification of homotrimers. We find that forces between helices in fibers from type I (as well as type II) homotrimers are not sensitive to pH between pH 6 and 7.5, in contrast to type I heterotrimers. Apparently, the effect of pH is related to extra histidine residues present on alpha2 chains but not on alpha1 chains. Finally, our measurements indicate that the alpha2 chain is responsible for binding some soluble compound(s), possibly glycosaminoglycans, whose displacement results, e.g., in the loss of tendon crystallinity. The ability of the alpha2 chain to bind non-collagen matrix components may be particularly important for bone matrix formation and mineralization. PMID- 11397099 TI - The allosteric activator Mg-ATP modifies the quaternary structure of the R-state of Escherichia coli aspartate transcarbamylase without altering the T<-->R equilibrium. AB - The allosteric enzyme aspartate transcarbamylase from Escherichia coli (ATCase) displays regulatory properties that involve various conformational changes, including a large quaternary structure rearrangement. This entails a major change in its solution X-ray scattering curve upon binding substrate analogues. We show here that, in the presence of the nucleotide effector ATP, known to stimulate the enzyme activity, the scattering profiles show a marked dependence on the metal bound to ATP. Whereas ATP has no major effect on the scattering pattern of ATCase, a saturating concentration of Mg-ATP notably modifies the scattering profile of the enzyme, either in the absence or in the presence of the bisubstrate analogue N-(phosphonacetyl)-l-aspartate (PALA). The transition with PALA in the presence of this metal-nucleotide complex remains concerted. Furthermore, Mg-ATP, as already observed with ATP, has no detectable direct effect on the T to R transition. The experimental scattering curves in the presence of Mg-ATP were fitted by a modeling approach using rigid body movements of the regulatory subunits and the catalytic trimers in the crystal structures. While the differences observed in the T-state in the presence of Mg-ATP are essentially attributed to the binding per se of the nucleotide, the solution structure of the R-state complexed to Mg-ATP is even more extended along the 3 fold axis than the previously described R solution structure, which is already more stretched out along the same axis than the crystal R structure. Based on the crystal structure of the enzyme in the R-state complexed with free ATP, a proposal is made to account for the effect of magnesium. PMID- 11397100 TI - Theoretical Study of the Electronic States of the Rb(2) Molecule. AB - We have calculated the electronic states of Rb(2) by multireference configuration interactions using the averaged relativistic effective small-core potential and the core-polarization potential. The potential energy curves for a large number of states dissociating into from 5s+5s up to 7s+5s asymptotic limits are calculated and the spectroscopic constants are reported. The spin-orbit effects for the states dissociating into 5p+5s and 4d+5s are calculated using the effective spin-orbit potential. The results are compared with available experimental data and other theoretical works. Copyright 2001 Academic Press. PMID- 11397101 TI - Simultaneous Analysis of the Rovibrational Levels upsilon(4)=1, 2, and 3 of HCCI and DCCI Based on Infrared Spectra. AB - The energy levels upsilon(4)=1, upsilon(4)=2, and upsilon(4)=3 of DCCI have been analyzed by using the fundamental nu(4)(1) (470-520 cm(-1)) and the overtone 2nu(4)(0) (955-1005 cm(-1)) bands together with the hot bands 2nu(4)(0,2)<- nu(4)(1), 3nu(4)(1,3)<--2nu(4)(0,2), and 3nu(4)(1)<--nu(4)(1). In the case of HCCI the previously studied hot bands connected to nu(4) have been completed by adding 3nu(4)(1)<--nu(4)(1) into the analysis. The various l-type resonances have been taken into account in the analyses of both the isotopomers. Furthermore, the Coriolis resonance between the close-lying nu(3) and nu(4) of DCCI has been considered. Altogether, accurate values for the molecular constants and the resonance parameters have been obtained from simultaneous analysis and they have been compared with those from separate analyses for different levels. Copyright 2001 Academic Press. PMID- 11397102 TI - Evaluation of Expansion Coefficients from Optimal Fitting Parameters for the Analysis of Spectra of Diatomic Molecules and an Application to LiH. AB - To evaluate individual expansion coefficients composing fitting parameters of the Born-Oppenheimer corrections to Dunham's coefficients Y(ij) that have been given analytically with the Delta(B) and Delta(omega) formalism, we examined the consistency of analytic expressions for those corrections with Watson's assertion of the experimental inseparability of nonadiabatic corrections Q(a, b)(r) for a molecule AB. Derived analytic expressions in terms of optimal fitting parameters for the corrections are essential to evaluate individual expansion coefficients. These expressions also reveal redundancies between empirical correction parameters Delta(ij). A method of evaluating nonadiabatic vibrational corrections Q(a, b)(r) and adiabatic corrections S(a, b)(r) separately consistent with Watson's assertion of inseparability is presented and is applied to an analysis of spectral data of LiH. Functions Q(a, b) and S(a, b) for LiH are thus successfully evaluated; S(H, Li)(r) values agree well with those predicted simply by wobble-stretch theory. Experimental values for optimal fitting parameters r(H)(1q) and r(H)(2q) are nearly equal to those of r(Li)(1q) and r(Li)(2q), respectively, in agreement with a theoretical relation r(a)(iq)=r(b)(iq). Copyright 2001 Academic Press. PMID- 11397103 TI - Hyperfine Parameters for Aluminum Hydride: An ab Initio Molecular Orbital Study. AB - An extensive ab initio molecular orbital study of the (27)Al nuclear spin rotation and nuclear quadrupolar coupling constants in aluminum hydride, AlH, has been performed. The (27)Al nuclear spin-rotation constant (C( perpendicular)), calculated to be approximately 300 kHz, was neglected in a previous analysis of the hyperfine structure in the microwave spectrum (M. Goto and S. Saito, Astrophys. J. 452, L147-148 (1995)). Unfortunately, the ab initio calculations do not provide a definitive value for the aluminum nuclear quadrupolar coupling constant, but suggest a value of -49+/-4 MHz. It is apparent that the microwave study of AlH should be repeated. Copyright 2001 Academic Press. PMID- 11397104 TI - The Ultraviolet Spectrum of the FeF Radical: A Study of the (6)Pi-X(6)Delta and (6)Phi-X(6)Delta Band Systems at 330 and 323 nm Respectively. AB - Two band systems of FeF have been recorded at Doppler-limited linewidths by laser induced fluorescence. The 330-nm system provided data for three subbands of the (0,0) band of the (6)Pi-X(6)Delta system and showed that levels of the (6)Pi(5/2) and (6)Pi(3/2) components are significantly perturbed. In the 323-nm system, the (0,0) band of the (6)Phi-X(6)Delta system was found to be overlapped by the (1,0) band of the (6)Pi-X(6)Delta system. Data were obtained for three subbands of the (1,0) (6)Pi-X(6)Delta system and for five subbands of the (0, 0) (6)Phi-X(6)Delta system. This work considerably extends the previous study by Pouilly et al. (B. Pouilly, J. Schamps, D. J. W. Lumley, and R. F. Barrow, J. Phys. B 11, 2281-2287 (1978)) because it is a higher resolution study with a lower temperature production of FeF by a continuous flow discharge method. The data recorded were combined with millimeter-wave data for FeF in the X(6)Delta state and fitted with an effective Hamiltonian to determine the major parameters for FeF in the X(6)Delta, (6)Pi, and (6)Phi electronic levels. Copyright 2001 Academic Press. PMID- 11397105 TI - Determination of Interatomic Potentials for the X0(+), A0(+), and B1 States of HgKr from Fluorescence and Excitation Spectra. AB - We present an analysis of the A0(+)(6(3)P(1))-->X0(+)(6(1)S(0)) bound-bound and bound-free fluorescence spectrum, and of the A0(+)(6(3)P(1))<--X0(+)(6(1)S(0)) and B1(6(3)P(1))<--X0(+)(6(1)S(0)) bound-bound excitation spectrum of the HgKr van der Waals molecule. The A-->X fluorescence spectrum, which was observed for the first time, as well as the excitation spectra were recorded using a pulsed supersonic molecular beam crossed with a pulsed dye laser beam. An analysis of the A(v')<--X(v"), B(v')<--X(v"), and A(v'=8)-->X(v") bound-bound bands indicates that a Morse function combined with a long-range approximation represents the interatomic potential energy curve of the A, B, and X states below the dissociation limit. In the simulation of the A(v'=8)-->X bound-free spectrum the Morse, Lennard-Jones (n-6), and Maitland-Smith (n(0), n(1)) functions were tested, and the Maitland-Smith (11.39, 10.50) potential was found to be a good representation of the repulsive part of the X-state PE curve above the dissociation limit, over the internuclear separation range R=2.85-3.55 A. The spectroscopic characteristics for the A, B, and X states obtained in this work are compared with other available experimental and theoretical results. Copyright 2001 Academic Press. PMID- 11397106 TI - Analysis of the nu(12) Band of Ethylene-(13)C(2) by High-Resolution FTIR Spectroscopy. AB - The Fourier transform infrared (FTIR) spectrum of the nu(12) fundamental band of ethylene-(13)C(2) ((13)C(2)H(4)) was recorded with an unapodized resolution of 0.004 cm(-1) in the frequency range from 1380 to 1500 cm(-1). Rovibrational constants for the upper state (nu(12)=1) up to five quartic and three sextic centrifugal distortion terms were derived for the first time by assigning and fitting a total of 1177 infrared transitions using a Watson's A-reduced Hamiltonian in the I(r) representation. The rms deviation of the fit was 0.00045 cm(-1). The ground state rovibrational constants were also determined for the first time by a fit of 738 combination differences from the present infrared measurements, with a rms deviation of 0.00060 cm(-1). The A-type nu(12) band with a band center at 1436.65411+/-0.00005 cm(-1) was found to be relatively free from local frequency perturbations. The inertial defect Delta(12) was found to be 0.24300+/-0.00002 uA(2). Copyright 2001 Academic Press. PMID- 11397107 TI - Torsional Splitting in the nu(5) Fundamental Infrared Band of CH(3)CD(3) and (13)CH(3)CD(3). AB - The nu(5) fundamental (C-C stretching) of CH(3)CD(3) shows a resolved torsional structure, caused by perturbations due mainly to the linear dependence of the torsional potential barrier on the normal coordinate Q(5). We were able to analyze this structure and to assign vibration-rotation transition wavenumbers for all five torsional components, classified according to the symmetry species of the G(18)((3)) extended molecular group. The torsional splitting pattern is qualitatively similar to that of a nondegenerate vibrational state with an even number of excited torsional quanta v(6). Explorative calculations show that the main perturber system should consist of the torsional components of the vibrational ground state correlating with v(6)=4 in the high barrier limit. The strength of the perturbation on the E(r0) torsional components of nu(5) increases rapidly with r, the E(40) component being the most affected. The observed transition wavenumbers can be reasonably fitted by a simplified model containing independent effective vibration-rotation parameters for the five different torsional components of nu(5), for both CH(3)CD(3) and (13)CH(3)CD(3). The trend of the determined values of the effective vibrational wavenumbers and rotational parameters over the torsional components supports the proposed vibration-torsion interaction mechanism, responsible for the observed torsional splittings. A strong anomaly observed in the rotational intensity distribution of nu(5) is discussed. Copyright 2001 Academic Press. PMID- 11397108 TI - The Rotational Spectra, Structure, Internal Dynamics, and Electric Dipole Moment of the Argon-Ketene van der Waals Complex. AB - Pulsed-beam Fourier transform microwave spectroscopy was used to observe and assign the rotational spectra of the argon-ketene van der Waals complex. Tunneling of the hydrogen or deuterium atoms splits the a- and b-type rotational transitions of H(2)CCO-Ar, H(2)(13)CCO-Ar, H(2)C(13)CO-Ar, and D(2)CCO-Ar into two states. This internal motion appears to be quenched for HDCCO-Ar where only one state is observed. The spectra of all isotopomers were satisfactorily fit to a Watson asymmetric top Hamiltonian which gave A=10 447.9248(10) MHz, B=1918.0138(16) MHz, C=1606.7642(15) MHz, Delta(J)=16.0856(70) kHz, Delta(JK)=274.779(64) kHz, Delta(K)=-152.24(23) kHz, delta(J)=2.5313(18) kHz, delta(K)=209.85(82) kHz, and h(K)=1.562(64) kHz for the A(1) state of H(2)CCO-Ar. Electric dipole moment measurements determined &mgr;(a)=0.417(10)x10(-30) C m [0.125(3) D] and &mgr;(b)=4.566(7)x10(-30) C m [1.369(2) D] along the a and b principal axes of the A(1) state of the normal isotopomer. A least squares fit of principal moments of inertia, I(a) and I(c), of H(2)CCO-Ar, H(2)(13)CCO-Ar, and H(2)C(13)CO-Ar for the A(1) states give the argon-ketene center of mass separation, R(cm)=3.5868(3) A, and the angle between the line connecting argon with the center of mass of ketene and the C=C=O axis, θ(cm)=96.4 degrees (2). The spectral data are consistent with a planar geometry with the argon atom tilted toward the carbonyl carbon of ketene by 6.4 degrees from a T-shaped configuration. Copyright 2001 Academic Press. PMID- 11397109 TI - The (1)(3)Pi Electronic State of LaF. AB - Rotational studies of bands of the infrared systems (1)(3)Pi-->(1)(3)Delta and (1)(3)Pi(1)-->X(1)Sigma(+) of lanthanum monofluoride were carried out with the aim of characterizing the (1)(3)Pi state, assuming the previous representations of X(1)Sigma(+) and (1)(3)Delta (1-3). The vibrational levels of (1)(3)Pi involved in the analyzed transitions were v=0, 1, 2 of the Omega=0 component, v=0, 1 of the Omega=1 component, and v=0 of the Omega=2 component. Perturbations were observed in both systems which were ascribed to spin-uncoupling interactions between (1)(3)Pi(2)(v=0) and (1)(3)Pi(1)(v=1) levels. A 9x9 matrix representation at equilibrium of the complex of interacting levels (v=0, 1, 2) was constructed, each diagonal v-block corresponding to a triplet model of the rovibrational (3)Pi Hamiltonian. The wavenumbers of some 1910 lines in the two systems were simultaneously fitted (rms error of the fit of about 0.0053 cm(-1)), thus yielding a consistent set of accurate spectroscopic constants for the (1)(3)Pi state; the spin-uncoupling interaction parameter was determined as B(Pi)(0, 1)=0.010917(13) cm(-1). Copyright 2001 Academic Press. PMID- 11397110 TI - High-Resolution Infrared, Microwave, and Submillimeter-Wave Study of FClO(2) in the nu(2), nu(3), nu(4), and nu(6) Excited States. AB - A high-resolution analysis of the {nu(2), nu(3)} and {nu(4), nu(6)} bands of the two isotopomers of chloryl fluoride F(35)ClO(2) and F(37)ClO(2) has been carried out for the first time using simultaneously infrared spectra recorded around 16&mgr;m and 26&mgr;m with a resolution of ca. 0.003 cm(-1) and microwave and submillimeter-wave transitions occurring within the vibrational states 2(1), 3(1), 4(1), and 6(1). Taking into account the Coriolis resonances which link the rotational levels of the {2(1), 3(1)} and the {4(1), 6(1)} interacting states, it was possible to reproduce very satisfactorily the observed transitions and to determine accurate vibrational energies and rotational constants for the upper states 2(1), 3(1), 4(1), and 6(1) of both the (35)Cl and (37)Cl isotopic species. Copyright 2001 Academic Press. PMID- 11397111 TI - Experimental and ab Initio Equilibrium Structure and Harmonic Force Field of 1,2,5-Oxadiazole. AB - The equilibrium structure of 1,2,5-oxadiazole has been calculated ab initio at the CCSD(T) level using a polarized valence quadruple zeta basis set. The harmonic force field has also been calculated at the MP2/cc-pVTZ, B3LYP/6 311++G(3df, 2pd), and B3LYP/cc-pVQZ levels. These force fields have been subsequently scaled and further refined by fitting them to the experimental values of the vibrational fundamentals of three isotopomers and the centrifugal distortion constants of the parent molecule. The specific refinement of those scaled force constants particularly sensitive to the experimental data set was decisive for obtaining a more reliable harmonic potential. The resulting force fields are presented and used, together with the ground state rotational constants, to calculate an r(z) structure. The experimental r(0), r(s), and r(m) structures have also been determined. The different results have been compared and it is concluded that the ab initio structure is a good approximation of the equilibrium structure. It is also shown that the magnetic correction is not negligible, particularly for the inertial defect. Another interesting conclusion is that the anharmonicity of the C-H stretching might be unusually small. Copyright 2001 Academic Press. PMID- 11397112 TI - Microwave Spectrum of the FCO Radical in the (2)A' Electronic Ground State. AB - The 1(01)-0(00), 2(02)-1(01), and 3(03)-2(02) rotational transitions of the FCO radical are observed at 22.3, 44.5, and 66.8 GHz, respectively, using a Fourier transform millimeter-wave spectrometer with a pulsed discharge nozzle. The FCO radical is produced by discharging F(2)CO diluted in the Ar buffer gas. Twelve fine and hyperfine components for the three transitions are observed, and the effective rotational constant, the centrifugal distortion constant, the spin rotation constant with its centrifugal correction term, and three hyperfine constants are determined. Furthermore, the vibrational satellites for the v(1)=1, v(2)=1, v(3)=1, and v(3)=2 states are also observed in the 22.3 GHz region. From the dipolar interaction constants, the principal axis of the dipolar interaction tensor is estimated and is discussed in relation to the distribution of the unpaired electron. Copyright 2001 Academic Press. PMID- 11397113 TI - Internal Rotation of the Hydroxyl Group in Isopropanol and the Chirality of the Gauche Form: Fourier Transform Microwave Spectroscopy of (CH(3))(2)CHOD. AB - The rotational spectrum of the deuterated isopropanol (CH(3))(2)CHOD has been observed by Fourier transform microwave spectroscopy and analyzed to yield tunneling splitting of 4431.4613 (17) MHz, between the antisymmetric and the symmetric gauche forms, which is much larger than the 2400 MHz estimated from the internal-rotation potential function reported in the literature. The potential function for the OH internal rotation has been examined in view of the discrepancy between the observed and estimated tunneling splitting, and it was accounted for by taking into account isotope effects on the potential constants. The deuterium quadrupole coupling effect has been included together with the Coriolis terms in the off-diagonal block of the Hamiltonian matrix for the gauche form. The deuterium quadrupole coupling constants obtained for the trans form were employed to calculate the components of the coupling constants as functions of the internal-rotation angle, and the components at around 120 degrees were compared with the values observed for the gauche form, thereby leading to unambiguous determination of the signs of the constants in the off-diagonal block; the signs are not obtainable from an ordinary analysis of the rotational spectra. The chirality of the gauche form was discussed by placing special emphasis on the effect of intermolecular interactions between two chiral molecules. Copyright 2001 Academic Press. PMID- 11397114 TI - Calculated Half-Widths and Line Shifts of Water Vapor Transitions in the 0.7 &mgr;m Region and a Comparison with Published Data. AB - Pressure-broadened half-widths and pressure-induced line shifts for the two most important bands of water vapor in the 0.7-&mgr;m region are determined using the complex Robert-Bonamy (CRB) formalism. The calculations are made with nitrogen and oxygen as the perturbing gas from which values for air as the perturbing gas are determined. The intermolecular potential is taken as a sum of electrostatic contributions and Lennard-Jones (6-12) atom-atom, and isotropic induction and dispersion components. The dynamics of the collision process are correct to second order in time. The calculated values are compared with published measurements and agreement is observed for both half-widths and line shifts. The temperature dependence of the half-width, which is necessary for reduction of remotely sensed data, is determined. Copyright 2001 Academic Press. PMID- 11397115 TI - Spectroscopy and Intensity Measurements of the 3nu(1)+3nu(3) Tetrad of (12)CO(2) and (13)CO(2). AB - Three of the four components of the 3nu(1)+3nu(3) tetrad of (12)C(16)O(2) and (13)C(16)O(2), labeled 30031, 30032, and 30033 in HITRAN notation, have been observed by intracavity laser absorption spectroscopy in the 10 450- to 11 000 cm(-1) region. The rotational analysis has yielded the rovibrational parameters of the vibrational states. The experimental values are found to be in very good agreement with the rovibrational energies recently predicted from variational calculations and reduced effective Hamiltonians. The absolute band intensity of these extremely weak transitions have been measured. The study of the relative intensities within the 3nu(1)+3nu(3) tetrad suggests that part of the oscillator strength is carried by the (22(0)3) state. Copyright 2001 Academic Press. PMID- 11397116 TI - Accurate Rovibrational Spectroscopic Properties of Cyanogen and Its Isotopomers. AB - The six-dimensional potential energy surface (PES) of the most stable isomer of C(2)N(2) is investigated using the coupled cluster single and double approach with perturbative treatment of triple excitations (CCSD(T)). The full quartic force fields of this molecule and some of its isotopomers are calculated in dimensionless normal coordinates. This quartic force field has been optimized in order to reproduce the known overtones within 1 cm(-1). A set of spectroscopic constants is derived for these isotopomers using this optimized quartic force field. The full rovibrational spectrum is also given up to 1500 cm(-1) with an accuracy better than 1 cm(-1). Copyright 2001 Academic Press. PMID- 11397117 TI - Calculated Vibrational Intensities in the A-&Xtilde; Electronic Transition of Acetylene. AB - Relative intensities in the vibrational structure of the A(1)A(u)(C(2h)) &Xtilde;(1)Sigma(+)(g)(D(infinityh)) electronic transition of acetylene are calculated. The calculation takes account of the large change of geometry and the change in the normal coordinates (the Dushinskii effect) between the two states. Because conventional vibrational wavefunctions for a nonlinear state do not behave correctly at linear geometries the vibrational integrals are only evaluated approximately. The transition ((1)Sigma(u)(-)-(1)Sigma(+)(g)) is forbidden at linear geometries, and so calculations are performed without and with a factor proportional to the angle of bend from linearity. Good agreement with experiment is obtained for the first few quanta of the absorption spectrum, lending support to the A-state harmonic potential of Tobiason et al., J. Chem. Phys. 99, 5762 (1993). Qualitative agreement is obtained for the emission spectrum (Jacobson and Field, J. Phys. Chem. 104, 3073 (2000)) when allowance is made for anharmonicity in the &Xtilde; state, but more quantitative agreement will require improvements in the empirical potential of the &Xtilde;-state, or an ab initio potential to higher energies than available at present. Copyright 2001 Academic Press. PMID- 11397118 TI - The 7-0 Overtone Spectrum of HCl. PMID- 11397119 TI - DiRef, A Database of References Associated with the Spectra of Diatomic Molecules. PMID- 11397120 TI - Factors affecting choice of surgical residency training program. AB - BACKGROUND: A significant problem facing American surgery today is the lack of participation from women and minorities. In 1995 and 1996, 15.1 and 15.8% of United States general surgical residency graduates were women. Of our 71 graduates in the last 12 years, 38% were women. The aim of this study was to identify the factors influencing our residents' choice of training program and the reasons why our program has a high percentage of female graduates. METHODS: Between 1989 and 2000, 27 women and 44 men completed general surgical training at our university and 44/71 (59%) responded to our survey. The age at residency completion was 34 +/- 2.2 years for men and 33.9 +/- 2.8 years for women. Fifty five percent of men and 30% of women went on to fellowship training; and 36% of men and 20% of women are in academia. RESULTS: Factors influencing our graduates' selection of training program are: Only 23% of men had a female faculty as their mentor, whereas 90% of women had a male faculty as their mentor during training. Only 59% of men but 80% of women (P < 0.05) agreed that female medical students need role models of successful female faculty members. Fifty-five percent of men and 45% of women would encourage a female medical student to choose surgery as a career, but 82% of men and 50% of women would encourage a male medical student to do so. Ninety-one percent of men and 85% of women would choose surgery as a career again. CONCLUSIONS: A surgical residency training program with strong leadership, good clinical experience, and high resident morale will equally attract both genders. Women may pay more attention to the program's gender mix and geographic location. PMID- 11397121 TI - NY-ESO-1 and CTp11 expression may correlate with stage of progression in melanoma. AB - BACKGROUND: As tumor cells spread beyond their primary site they undergo changes in their gene expression that may be detectable and useful for microstaging. The cancer/testis (CT) antigens are a family of proteins that include MAGE 1-3, NY ESO-1, SSX 1-5, and others that are potential markers for microstaging melanoma. CT antigens are produced by many tumor types but few normal tissues other than testis. One CT antigen, CTp11, was shown to be expressed by metastasizing melanoma cell lines but not by nonmetastasizing variants. We tested this finding by studying the expression of CTp11 and NY-ESO-1 by melanoma samples from different stages of progression. MATERIALS AND METHODS: We collected 20 primary, 22 locoregional, and 10 distant metastatic melanoma samples. We extracted total RNA, and reverse transcription yielded cDNA, which was PCR-amplified using primers to detect beta-actin, tyrosinase, MART-1, NY-ESO-1, and CTp11. The PCR products were separated on ethidium bromide-stained agarose gels and visualized by UV transillumination. RESULTS: All samples were positive for beta-actin and MART-1 and all but two were positive for tyrosinase, confirming RNA integrity and the presence of melanoma. Twenty-seven samples were positive for NY-ESO-1, CTp11, or both. CTp11 tended to be expressed by primary melanomas and NY-ESO-1 by metastatic samples (P < 0.02). CONCLUSIONS: There is a statistically significant difference in the distribution of the expression of CTp11 and NY-ESO-1 in melanoma from different stages of progression. NY-ESO-1 may be a marker of more advanced disease and CTp11 of less advanced disease. PMID- 11397122 TI - Quality control of resident operative experience: compliance with RRC criteria. AB - PURPOSE: Process Capability Analysis (PCA) is a quality control tool that can be applied to how resident cases are allocated. PCA measures how close an output is to its target (Cp) and its location (K) in relation to Cp. For resident cases, the statistics of Cp and K measure variability in case numbers, which is useful in planning how a program meets RRC criteria in operative experience on specialty services. METHODS: A review of 6 years of vascular surgery cases (1994-1999) using PG 5 RRC case logs and the departmental quality database as sources was done. PCA was applied to resident case numbers. RRC 1997-1998 National Program Data were used to define quality control limits. The 30th, 50th, and 70th percentiles in vascular procedures served as lower, nominal, and upper control limits, respectively. RESULTS: Cases were grouped into aortic (AO), cerebrovascular (CER), peripheral (PER), and visceral (VSC), to meet RRC definition for major reconstructions. PCA analysis of PG5 RRC submitted data, total cases reported by the graduating residents (n = 20) in their submitted RRC Case logs, and department QA log numbers over 6 years of the period revealed the following mean numbers for AO = 13.7 +/- 4.2 (Cp = 0.279, K = +70.5%), CER = 15.6 +/- 6.4 (Cp = 0.365, K = +78.6%), PER = 23.1 +/- 10.5 (Cp = 0.237, K = +80.1%), VSC = 2.5 +/- 1.5 (Cp = 0.189, K = +175.0%), and total = 55.1 +/- 20.2 (Cp = 0.305, K = +70.5%). CONCLUSIONS: Processes that are in statistical control should have a Cp > 1.0, and a small K: i.e., centered tightly and near the desired nominal limit. By PCA analysis, the vascular surgery service product of PG5 case numbers is not in statistical control, but instead exceeds the defined control limits (RRC 50th percentile levels). The low Cp values demonstrate that specific case numbers may vary from resident to resident. The high positive K percentages imply that more residents have case numbers that meet or exceed RRC 50th percentiles. Sources of potential error and variability may be partially explained by the data of total case volume and resident underreporting. More accurate RRC reporting by residents could account for up to 20% more cases, potentially offsetting individual case allocation differences within a particular resident year. PMID- 11397123 TI - Heterotypic smooth muscle cell/endothelial cell interactions differ between species. AB - BACKGROUND: In vitro coculture models have been used to study heterotypic cell cell interactions. This study was performed to determine if species of cell origin affects heterotypic smooth muscle cell (SMC) endothelial cell (EC) interactions in coculture. METHODS: To study the effect of ECs on SMC proliferation, ECs were cultured on porous Dacron membranes. SMCs were added opposite the ECs or on bare membranes on Day 3, and after 4 days, cells were harvested for cell counts. To study the effect of SMCs on EC proliferation, ECs at a density of 5 x 10(5) cells/membrane were added to bare membranes or on membranes opposite SMCs plated 2 days earlier. After 48 h, cells were harvested for cell counts. (N = 3/condition, experiments repeated x2.) Cells of human and bovine aortic origin were used. RESULTS: The effect of coculture on cell growth differed between species. The effect of heterotypic interactions between human cocultured cells was coinhibitory on the rate of growth as compared to the growth of cells cultured alone. Growth of cocultured ECs was 55.2 +/- 8.7% less than that of ECs cultured alone while growth of cocultured SMCs was 27.2 +/- 6.0% less than growth of SMCs cultured alone. This contrasted with the bovine EC stimulation of SMC proliferation, with 66.8 +/- 5.0% greater growth of cocultured SMCs compared to SMCs cultured alone, and failure of bovine SMCs to decrease EC proliferation. CONCLUSIONS: Since significant differences in cell-material interactions occur in vivo between species, the finding that in vitro heterotypic cell-cell interactions are species dependent is not surprising. This fundamental difference in cell behavior stresses the potential importance of using human cells in studies evaluating cell-cell and cell-material interactions in vitro. PMID- 11397124 TI - Environmental factors of temperature, humidity, serum accumulation, and cell seeding increase colon cancer cell adhesion in vitro, with partial characterization of the serum component responsible for pressure-stimulated adhesion. AB - Physical characteristics of surgical wounds and viable tumor cells shed may differ between open and laparoscopic procedures. Because environmental factors may vary between the laparoscopic milieu and that of open surgical procedures, we sought to characterize the effect of these factors on tumor cell adhesion, an early step in the process of wound implantation. Human SW620 colon cancer cells were placed in matrix-precoated dishes for 30 min at concentrations of 90,000 540,000 cells/well, at 25-37 degrees C, in the native state of the matrix proteins and after drying for 60 min, and in 0-10% serum. As increased pressure has previously been reported to stimulate colon cancer cell adhesion synergistically with serum, we then further partially characterized the serum components responsible for this potentiating effect. The number of adherent cells varied linearly with cells seeded. Adhesion was temperature-dependent, and also was dependent on the matrix conformation. Less adhesion occurred to dry matrix proteins. Serum dose-dependently potentiated SW620 pressure-stimulated adhesion, with a maximal increase in adhesion compared with ambient pressure conditions at 5% serum concentration. Heat inactivating the serum at 60 degrees C for 30 min ablated the effect. Filtration to remove molecules over 10 kDa produced no change in adhesion relative to ambient conditions, but filtration to 100 kDa preserved the serum effect. When the serum was passed over a gelatin-Sepharose column, which binds numerous proteins including fibronectin, the serum effect was lost. Addition of fibronectin to serum-free media did not reconstitute the effect. The environmental factors of warm temperature, moisture, and serum accumulation may contribute to increased colon cancer cell adhesion. However, the most important determinant of malignant adhesion to surgical wounds, laparoscopic or open, is likely to be the size of the tumor cell inoculum. Pressure stimulation of colon cancer cell adhesion is potentiated by heat-labile serum components of molecular weight 10-100 kDa which bind gelatin-Sepharose, and is not fibronectin alone. Irrigating serum from surgical wounds may decrease tumor implantation. PMID- 11397125 TI - The impact of a clinical pathway for gastric bypass surgery on resource utilization. AB - BACKGROUND: Clinical pathways are believed to improve patient care and reduce costs. Our hypothesis was that a gastric bypass pathway would decrease hospital resource utilization and cost of care without adversely affecting patient care. METHODS: The prepathway (Pre) group consisted of 16 gastric bypasses (6/98 to 3/99). The postpathway (Post) group includes 12 gastric bypass procedures performed after institution of the clinical pathway (4/99 to 12/99). The impact of the clinical pathway on hospital length of stay (LOS) and resource utilization was investigated. A comparison of costs was performed using cost/charge ratios. Hospital readmissions and postoperative complications were also examined. RESULTS: Despite increased obesity/medical acuity of the Post group, hospital LOS decreased by 3 days (P < 0.0001). Total hospital costs decreased by over $1600/case (>15%). Postpathway savings were greatest for room and board (34%), supplies (41%), and lab/radiology costs (50%). An increase in OR costs (22%) was observed in the Post group. This was due to an increase in anesthesia time (epidural catheter placement) and equipment costs (ultrasonic shears). Despite reductions in hospital LOS and resource utilization, the complication rate (Pre 12%, Post 16%) was similar and two patients in each group required brief readmission. CONCLUSIONS: A pathway for gastric bypass decreased hospital LOS and resource utilization. OR-related expenses account for 34-50% of total costs and must be monitored closely for surgical patients. The reduction in costs observed with this clinical pathway was not associated with an increase in postoperative complications or hospital readmission. PMID- 11397126 TI - Bryostatin/ionomycin-activated T cells mediate regression of established tumors. AB - We have shown that adoptive transfer of tumor-sensitized lymphocytes activated in vitro with bryostatin-1 and ionomycin (B/I), and expanded in culture, can induce regression of small established tumors. We set out to determine whether similar treatment would be effective against larger tumors and what cells mediate this effect. We also attempted to shorten the ex vivo culture period with the ultimate aim of developing a more clinically useful protocol. BALB/c mice were injected in one footpad with IL-2-transfected 4T07 mammary tumor cells. Ten days later, popliteal draining lymph nodes (DLN) were harvested and activated with B/I for 18 h. Mice with either 3-day or 10-day 4T07 flank tumors were treated with cyclophosphamide (100 mg/ kg ip, CYP) alone or CYP followed the next day by infusion of either B/I-activated lymphocytes transferred immediately or activated cells that had been expanded in vitro for 3 or 10 days. In some experiments, mice were also treated with rat anti-mouse CD4 monoclonal antibody (GK1.5) or anti-CD8 antibody (2.43). All mice receiving CYP alone or CYP + sensitized, nonactivated DLN cells demonstrated progressive tumor growth. One hundred percent (6/6) of mice treated with CYP + AIT with B/I-activated,10-day expanded cells had complete regression of 3-day flank tumors. Treatment with activated, nonexpanded cells, induced tumor regression in a majority of mice, but was not as reliable as AIT with expanded cells. We developed a protocol with a shortened expansion period (3 day) that was efficacious for treatment of 4T07 when adoptively transferred to either 3 or 10 day tumor-bearing mice. In vivo depletion of CD4(+) cells had no effect on regression of 3-day tumors, but treatment with anti-CD8 antibody abrogated the effect of immunotherapy. Adoptive transfer of B/I-activated cells, with or without long-term expansion, induced regression of early and late stage 4T07 tumors and is dependent on CD8(+) but not CD4(+) T cells. PMID- 11397127 TI - Integrin expression, enterocyte maturation, and bacterial internalization. AB - BACKGROUND: Little is known about the molecular mechanisms involved in the translocation of enteric bacteria. Adhesion molecules mediate interactions between some enteric pathogens and mammalian cells, but no such interactions have been identified for enterocytes and normal enteric bacteria. Using enteric pathogens, adhesion molecule expression has been linked to bacterial internalization and to enterocyte differentiation. Therefore, experiments were designed to study enterocyte integrin expression and differentiation, as well as enterocyte internalization of Salmonella typhimurium, Proteus mirabilis, and Escherichia coli. MATERIALS AND METHODS: Relative expression of the alpha2, alpha3, and beta1 integrin subunits on Caco-2 and HT-29 enterocytes (mature and immature) was measured by ELISA. Bacteria-enterocyte surface interactions were observed by light and scanning electron microscopy. Bacterial internalization by enterocytes was quantified using the gentamicin protection assay. RESULTS: Expression of the alpha2, alpha3, and beta1 integrin subunits was consistently increased in immature compared to mature Caco-2 enterocytes; however, compared to mature enterocytes, immature HT-29 enterocytes had similar expression of alpha3 and beta1 but decreased alpha2. Compared to untreated mature enterocytes, bacterial internalization was increased in immature enterocytes as well as mature enterocytes with lateral membranes artifactually exposed. However, there was no difference in bacterial internalization between immature enterocytes and mature enterocytes treated to expose the lateral membrane. CONCLUSIONS: Bacterial internalization by enterocytes appeared to be due to factors other than integrin expression or enterocyte differentiation. Exposure of the lateral enterocyte membrane may play an important role in facilitating bacterial internalization by enterocytes. PMID- 11397128 TI - Iron deficiency transiently suppresses biliary neuronal nitric oxide synthase. AB - BACKGROUND: Iron deficiency results in altered gallbladder and sphincter of Oddi (SO) motility and cholesterol crystal formation. In addition, gallbladder neuronal nitric oxide synthase (nNOS) has been shown to be markedly reduced after 8 weeks on an iron-deficient diet. However, the effects of prolonged iron deficiency on gallbladder and SO nNOS as well as crystal formation have not been determined. Therefore, we tested the hypothesis that iron deficiency would downregulate both gallbladder and SO nNOS expression and that nNOS downregulation and cholesterol crystal formation would progress over time. MATERIALS AND METHODS: Thirty-eight adult female prairie dogs were fed either an ironsupplemented (Fe+) (200 ppm) or an iron-deficient (Fe-) (8 ppm) diet for 8 weeks (Fe+ n = 9, Fe- n = 10) or 16 weeks (Fe+ n = 9, Fe- n = 10). Blood hemoglobin (HbG) was measured; gallbladder cholesterol crystals were counted; and cholesterol saturation indices (CSI) were calculated. Gallbladder and SO nNOS levels were measured by Western blot. RESULTS: The Fe+ prairie dogs had significantly higher HbG than the Fe- animals (16.9 +/- 0.6 g/dl vs 15.2 +/- 0.5 g/dl, respectively, P < 0.05) after 8 weeks. This difference was even greater after 16 weeks (16.1 +/- 0.4 g/dl vs 14.0 +/- 0.5 g/dl, P < 0.01). At 8 weeks, more cholesterol crystals per 10 HPF were observed in the Fe- animals (0.4 +/- 0.3 vs 1.6 +/- 0.4 per 10 HPF, P < 0.05). This difference was even greater after 16 weeks (0.0 +/- 0.0 vs 52.6 +/- 25.3 per 10 HPF, P < 0.01). No difference in the CSI was observed in the four groups. Iron deficiency decreased the nNOS/beta actin protein levels in the gallbladder and SO at 8 weeks (57.0 +/- 29.6 vs 7.4 +/- 2.6, gallbladder, P < 0.05) (98.4 +/- 39.7 vs 29.9 +/- 11.0, SO, P = 0.09), but these levels returned to baseline at 16 weeks. CONCLUSIONS: We conclude that iron deficiency acutely suppresses gallbladder and SO nNOS, and that compensatory mechanisms return nNOS to baseline levels while cholesterol crystal formation increases over time. PMID- 11397129 TI - Recycling probability and dynamical properties of germinal center reactions. AB - We introduce a new model for the dynamics of centroblasts and centrocytes in a germinal center. The model reduces the germinal center reaction to the elements considered as essential and embeds proliferation of centroblasts, point mutations of the corresponding antibody types represented in a shape space, differentiation to centrocytes, selection with respect to initial antigens, differentiation of positively selected centrocytes to plasma or memory cells and recycling of centrocytes to centroblasts. We use exclusively parameters with a direct biological interpretation such that, once determined by experimental data, the model gains predictive power. Based on the experiment of Han et al. (1995b) we predict that a high rate of recycling of centrocytes to centroblasts is necessary for the germinal center reaction to work reliably. Furthermore, we find a delayed start of the production of plasma and memory cells with respect to the start of point mutations, which turns out to be necessary for the optimization process during the germinal center reaction. The dependence of the germinal center reaction on the recycling probability is analysed. PMID- 11397130 TI - Effects of habitat destruction and resource supplementation in a predator-prey metapopulation model. AB - We developed a mean field, metapopulation model to study the consequences of habitat destruction on a predator-prey interaction. The model complements and extends earlier work published by Bascompte and Sole (1998, J. theor. Biol.195, 383-393) in that it also permits use of alternative prey (i.e., resource supplementation) by predators. The current model is stable whenever coexistence occurs, whereas the earlier model is not stable over the entire domain of coexistence. More importantly, the current model permits an assessment of the effect of a generalist predator on the trophic interaction. Habitat destruction negatively affects the equilibrium fraction of patches occupied by predators, but the effect is most pronounced for specialists. The effect of habitat destruction on prey coexisting with predators is dependent on the ratio of extinction risk due to predation and prey colonization rate. When this ratio is less than unity, equilibrial prey occupancy of patches declines as habitat destruction increases. When the ratio exceeds one, equilibrial prey occupancy increases even as habitat destruction increases; i.e., prey "escape" from predation is facilitated by habitat loss. Resource supplementation reduces the threshold colonization rate of predators necessary for their regional persistence, and the benefit derived from resource supplementation increases in a nonlinear fashion as habitat destruction increases. We also compared the analytical results to those from a stochastic, spatially explicit simulation model. The simulation model was a discrete time analog of our analytical model, with one exception. Colonization was restricted locally in the simulation, whereas colonization was a global process in the analytical model. After correcting for differences between nominal and effective colonization rates, most of the main conclusions of the two types of models were similar. Some important differences did emerge, however, and we discuss these in relation to the need to develop fully spatially explicit analytical models. Finally, we comment on the implications of our results for community structure and for the conservation of prey species interacting with generalist predators. PMID- 11397131 TI - Control mechanisms of diel vertical migration: theoretical assumptions. AB - We explore control mechanisms underlying the vertical migration of zooplankton in the water column under the predator-avoidance hypothesis. Two groups of assumptions in which the organisms are assumed to migrate vertically in order to minimize realized or effective predation pressure (type-I) and to minimize changes in realized or effective predation pressure (type-II), respectively, are investigated. Realized predation pressure is defined as the product of light intensity and relative predation abundance and the part of realized predation pressure that really affects organisms is termed as effective predation pressure. Although both types of assumptions can lead to the migration of zooplankton to avoid the mortality from predators, only the mechanisms based on type-II assumptions permit zooplankton to undergo a normal diel vertical migration (morning descent and evening ascent). The assumption of minimizing changes in realized predation pressure is based on consideration of DVM induction only by light intensity and predators. The assumption of minimizing changes in effective predation pressure takes into account, apart from light and predators also the effects of food and temperature. The latter assumption results in the same expression of migration velocity as the former one when both food and temperature are constant over water depth. A significant characteristic of the two type-II assumptions is that the relative change in light intensity plays a primary role in determining the migration velocity. The photoresponse is modified by other environmental variables: predation pressure, food and temperature. Both light and predation pressure are necessary for organisms to undertake DVM. We analyse the effect of each single variable. The modification of the phototaxis of migratory organisms depends on the vertical distribution of these variables. PMID- 11397132 TI - Slow proliferation as a survival advantage: an attempt to resolve a paradox. AB - A model is proposed for immunological high zone- and self-tolerance that is based on the concept of overstimulation of replication: It is demonstrated that if proliferation is hindered by delayed response to nutritional deficiency, then under competitive conditions slower proliferation can become a survival advantage. This offers a resolution of the paradox that self-antigens promote both positive and negative selection in the development of lymphocytes. It also suggests that maintaining nutrition and caloric intake at a minimal level can prolong the survival of cancer patients if the cancer cells replicate more rapidly than healthy ones. PMID- 11397133 TI - A parallel study on bacterial growth and inactivation. AB - Pure stochastic birth process models are used to establish the connection between the traditional food microbiology concept of bacterial lag period, before the exponential growth, and the distribution of the lag times of individual cells. In a parallel way, similar study is carried out on the connection between the "shoulder" period, before the exponential decay, and the distribution of the survival times of individual cells. Formulae are derived to calculate the parameters of the growth/survival curves from the distributions of the respective parameters of individual cells. It is shown that, in some aspects, analogy, in some aspects fundamental difference exists between growth and survival modelling. PMID- 11397134 TI - A model of the gastric gland ejection cycle: low ejection fractions require reduction of the glandular dead space. AB - This paper was inspired by the reported results of authors from Uppsala and Lund that gastric glands in rats rhythmically contract 3-7 cycles per minute and develop luminal pressures more than 10 mmHg. To ensure that pepsinogen is not retained in the acid-rich section of the gland, ejection fractions would need to be more than 50% of the gland volume. We have tried to calculate the ejection fraction of such contractions. Dimensions of human gastric glands were measured on the fresh frozen samples of macroscopically and histologically normal gastric mucosa. In total, 18 specimens (from nine persons) were measured under the microscope. The density of glands was 135 +/- 11 (mean +/- S.D.) glands per mm( 2) of gastric mucosa. A typical gastric gland is a tubular structure 1.2 +/- 0.22 mm long and 0.03-0.05 mm wide. We have used 1 mm for length and 0.03 mm for the gland diameter to calculate that each gland approximates a volume of 707 pl, suggesting that the total glandular volume for 15 million glands reaches 10.6 ml. Further calculations based on one to five contractions per minute on an average and on the total volume of gastric glands of 10 ml showed that only ejection fractions less than 10% deliver daily volumes less than 3 l. The presented model of the gastric gland activity is based on the idea that the low ejection fractions require a reduction of the glandular dead space. The reduced luminal pressure during the gland relaxation might cause backflux of hydrophobic viscoelastic mucus through the gland aperture. Repeated glandular contractions and relaxations would move the mucus all the way to the gland bottom, filling the gland cavity below the neck with an axial semisolid mucous cylinder. This filling would reduce the gland dead space. During contractions, the gland would eject mainly the peripheral, the more liquid part of its content. The decreasing luminal pressure in the relaxing gland would pull the outlet mucus inside, protecting gland apertures from the gastric juice. PMID- 11397135 TI - Handedness, birth weight, mortality and Barker's hypothesis. PMID- 11397136 TI - The influence of turbulence on plankton predation strategies. AB - The importance of predation in regulating the size of competing plankton and larval fish populations has long been appreciated. However, it has only recently been recognized that turbulence must have a significant influence on predator prey interactions because most rival species of microorganisms co-exist in oceanic or fast moving fresh water flows. Turbulence is likely to influence predation strategies in two ways. The extra energy imparted to a micro-organism from the flow field will enhance the number of encounters or "contacts" between predators and prey. At the same time, because the velocity of a predator relative to its potential prey will be increased, the time-scale over which a capture must be completed is reduced. Balancing the benefits of extra encounters with the drawbacks of more difficult captures, will dictate an optimal predation strategy, either foraging behaviour or ambush feeding, on the predator. This will depend on its own and the prey's swimming capabilities, as well as the characteristics of the turbulent environment. In this paper some previous work, examining the increased encounter rate in turbulence, will be extended to look at the capture problem. The main proposal is that the capture event should be encapsulated in a capture probability function, from which the optimal predation strategy can be derived. As an illustration, plausible capture probability functions will be postulated and the resulting predictions tested against numerical simulations carried out in a turbulent-like flow field. Good agreement between the predictions and the simulations is demonstrated. PMID- 11397137 TI - Thermodynamical implications of a protein model with water interactions. AB - We refine a protein model that reproduces fundamental aspects of protein thermodynamics. The model exhibits two transitions, hot and cold unfolding. The number of relevant parameters is reduced to three: (1) binding energy of folding relative to the orientational energy of bound water, (2) ratio of degrees of freedom between the folded and unfolded protein chain, and (3) the number of water molecules that can access the hydrophobic parts of the protein interior upon unfolding. By increasing the number of water molecules in the model, the separation between the two peaks in the heat capacity curve comes closer, which is more consistent with experimental data. In the end we show that if we, as a speculative assumption, assign only two distinct energy levels for the bound water molecules, better correspondence with experiments can be obtained. PMID- 11397138 TI - A computational model of dendrite elongation and branching based on MAP2 phosphorylation. AB - We introduce a new computational model of dendritic development in neurons. In contrast to previous models, our model explicitly includes cellular mechanisms involved in dendritic development. It is based on recent experimental data which indicates that the phosphorylation state of microtubule-associated protein 2 (MAP2) may play a key role in controlling dendritic elongation and branching (Audesirk et al., 1997). Dephosphorylated MAP2 favours elongation by promoting microtubule polymerization and bundling, whilst branching is more likely to occur when MAP2 is phosphorylated and microtubules are spaced apart. In the model, the rate of elongation and branching is directly determined by the ratio of phosphorylated to dephosphorylated MAP2. This is regulated by calmodulin dependent protein kinase II (CaMKII) and calcineurin, which are both dependent on the intracellular calcium concentration. Results from computer simulations of the model suggest that the wide variety of branching patterns observed among different cell types may be generated by the same underlying mechanisms and that elongation and branching are not necessarily independent processes. The model predicts how the branching pattern will change following manipulations with calcium, CaMKII and MAP2 phosphorylation. PMID- 11397139 TI - Aposematic (warning) coloration associated with thorns in higher plants. AB - Aposematic coloration, a well-known phenomenon in animals, has been given little attention in plants. Here I discuss two types of conspicuousness of thorns which are typical of many plant species: (1) colorful thorns, and (2) white spots, or white and colorful stripes, associated with thorns in leaves and stems. Both types of aposematic coloration predominate the spine system of taxa rich with spiny species-Cacti, the genera Agave, Aloe and Euphorbia. The phenomena have been recorded here in over a thousand species originating in several continents of both the Old and New World. I propose that this is a case of vegetal aposematic coloration analogous to such coloration of poisonous animals, and which communicates between plants and herbivores. PMID- 11397140 TI - Tensile fatigue in bone: are cycles-, or time to failure, or both, important? AB - In life, bones are subjected to fatigue loading which has different frequency and amplitude components, as well as various kinds of loading modes like tension, compression, shear and combinations of them. Considerable variability is observed in fatigue results of bone, which may be caused by these experimental variables or by the bone itself. In past studies the effect of magnitude and mode of loading have been examined in standard fatigue strength (stress vs. cycles to failure) diagrams. The effect of frequency is not clear, but there is clear evidence (from Carter & co-workers) that, at least in human bone, tension "fatigue" failure was determined solely by time rather than by cycles. We sought to confirm these results in the same and a different species. We cycled human and bovine bone in tension at two frequencies: 0.5 and 5 Hz. There was no cycle number effect; the results from the tests at the two frequencies were different if plotted and analysed as a function of cycles to failure, but were not separable if plotted and analysed as a function of time to failure. In this respect bone differs from tendon, in which failure in tension is a function of both cycles and time. PMID- 11397141 TI - Ab initio and molecular mechanics (MM3) calculations of protonated-neutral diamine hydrogen bonds. AB - Ab initio calculations of cation-neutral diamine complexes have been carried out at the MP2/6-311+G** level. The geometry and energetics of the charge-reinforced hydrogen bond are analyzed with respect to the alkyl substitution of both the protonated and neutral nitrogen atoms, and these results have been used to improve the quality of the MM3(2000) force field. In addition, specialized hydrogen bond parameters optimized for MM3(2000) are presented. These parameters allow very accurate gas-phase modeling of the charge-neutral diamine environment. Molecular mechanics calculations can model effectively protonated amine-neutral amine hydrogen bonds in the gas phase and solution (continuum dielectric) through a combination of charge-dipole interactions and explicit hydrogen-bonding terms. PMID- 11397142 TI - An immunotherapeutic program for the treatment of nicotine addiction: hapten design and synthesis. AB - People continue to smoke and use tobacco products despite well-established hazardous consequences. The most contributing factor is the addictive nature of nicotine. There is no highly effective treatment for the problem of nicotine dependence. Immunotherapy offers an alternative to conventional approaches. The chemistry necessary for a comprehensive immunopharmacological program is presented. Haptens for the generation of antibodies specific for naturally occurring (S)-nicotine, (S)- and (R)-nornicotine, and the metabolite (S)-cotinine were prepared with high optical purity. Preliminary data for antinicotine antibodies are reported. PMID- 11397143 TI - A theoretical study of alcohol oxidation by ferrate. AB - The conversion of methanol to formaldehyde mediated by ferrate (FeO(4)2-), monoprotonated ferrate (HFeO4-), and diprotonated ferrate (H2FeO4) is discussed with the hybrid B3LYP density functional theory (DFT) method. Diprotonated ferrate is the best mediator for the activation of the O-H and C-H bonds of methanol via two entrance reaction channels: (1) an addition-elimination mechanism that involves coordination of methanol to diprotonated ferrate; (2) a direct abstraction mechanism that involves H atom abstraction from the O-H or C-H bond of methanol. Within the framework of the polarizable continuum model (PCM), the energetic profiles of these reaction mechanisms in aqueous solution are calculated and investigated. In the addition-elimination mechanism, the O-H and C H bonds of ligating methanol are cleaved by an oxo or hydroxo ligand, and therefore the way to the formation of formaldehyde is branched into four reaction pathways. The most favorable reaction pathway in the addition-elimination mechanism is initiated by an O-H cleavage via a four-centered transition state that leads to intermediate containing an Fe-O bond, followed by a C-H cleavage via a five-centered transition state to lead to formaldehyde complex. In the direct abstraction mechanism, the oxidation reaction can be initiated by a direct H atom abstraction from either the O-H or C-H bond, and it is branched into three pathways for the formation of formaldehyde. The most favorable reaction pathway in the direct abstraction mechanism is initiated by C-H activation that leads to organometallic intermediate containing an Fe-C bond, followed by a concerted H atom transfer from the OH group of methanol to an oxo ligand of ferrate. The first steps in both mechanisms are all competitive in energy, but due to the significant energetical stability of the organometallic intermediate, the most likely initial reaction in methanol oxidation by ferrate is the direct C-H bond cleavage. PMID- 11397144 TI - Substituent influences on the stability of the ring and chain tautomers in 1,3 O,N-heterocyclic systems: characterization by 13C NMR chemical shifts, PM3 charge densities, and isodesmic reactions. AB - Substituent effects on the stabilities of the ring and chain forms in a tautomeric equilibrium of five series of 2-phenyloxazolidines or -perhydro-1,3 oxazines possessing nine different substitutions at the phenyl moiety have been studied with the aid of 13C NMR spectroscopy and PM3 charge density and energy calculations. Reaction energies of the isodesmic reactions, obtained from the calculated energies of formation, show that electron-donating substituents stabilize both the chain and ring tautomers but the effect is stronger on the stability of the chain form than on that of the ring form. The 13C chemical shift changes induced by the phenyl substituents (SCS) were analyzed by several different single and dual substituent parameter approaches. The best correlations were obtained by equation SCS = rhoFsigmaF + rhoRsigmaR. In all cases the rhoF values and in most cases also the rhoR values were negative at both the C=N and C 2 carbons, indicating a reverse behavior of the electron density. This concept could be verified by the charge density calculations. The 13C chemical shifts of the C=N and C-2 carbons show a normal dependence on the charge density (q(tot)), but the charge density shows a reverse dependence on substitution. Correlation analysis of the 13C chemical shifts, solvent effect (CDCl3 vs DMSO-d6) on the NMR behavior as well as the effect of substituents on the electron densities and on the stabilities of the ring and chain tautomers show that the substituent dependence of the relative stability of the ring and chain tautomers in equilibrium is governed by several different electronic effects. At least intramolecular hydrogen bonding between the imine nitrogen and the hydroxyl group as well as polarization of the C=N bond seem to contribute in the chain form. Stereoelectronic and electrostatic effects are possible to explain the increase in stability of the ring form by electron-donating substituents. PMID- 11397145 TI - Highly enantioselective synthesis of alpha,beta-diaminopropanoic acid derivatives using a catalytic asymmetric hydrogenation approach. AB - Rh-DuPhos-catalyzed asymmetric hydrogenation of alpha,beta-diamidoacrylates provides a highly efficient and enantioselective route to chiral alpha,beta diaminopropanoic acid derivatives. The mechanistic course of the hydrogenation was studied using isotopically enriched enamide complexes and phosphorus and carbon NMR. Addition of methyl alpha-N-benzoyl-beta-N-acetyl-diaminopropenoate to the solvated catalyst gave a single 1:1 enamide complex and demonstrated the binding of the olefin and alpha-amide carbonyl group; the carboxylate and beta-N acyl groups did not bind to the metal. Changes to the electronic and steric properties of the beta-N-acyl group were well tolerated; however, small changes to the binding alpha-N-acyl group were found to significantly affect hydrogenation yields. PMID- 11397146 TI - Expedient asymmetric synthesis of all four isomers of N,N'-protected 2,3 diaminobutanoic acid. AB - 2,3-Diaminobutanoic acid (DAB) is found in several peptide antibiotics, toxins, and biologically active molecules. This paper describes the practical and highly enantioselective synthesis of all four N,N'-protected DAB stereoisomers using an asymmetric Rh(I)-phosphine-catalyzed hydrogenation of isomeric enamides as the key step. Thermal and photochemical isomerization of the enamide hydrogenation substrates coupled with catalyst-geometric isomer pairing allows targeted synthesis of single DAB isomers in maximum yield. PMID- 11397148 TI - Characterization of iodonium salts differing in the anion. AB - The properties and reactivity of a series of iodonium salts with different anions were compared. The nucleophilicity of the anions in such compounds can be characterized by their melting points and NMR spectra. When using Quinaldine Red as indicator and CH3CN as solvent, the acid release rate of the iodonium salts correlated very well with their polymerization results in acid-sensitive epoxides. PMID- 11397147 TI - Tartric acid-based linker for the solid-phase synthesis of C-terminal peptide alpha-oxo aldehydes. AB - A novel linker, based on the anchoring of (+)-dimethyl 2,3-O-isopropylidene-D tartrate to PEGA or PEG-PS solid supports, was developed for the solid-phase synthesis of C-terminal peptide alpha-oxo aldehydes. Peptide elongation was performed using the 9-fluorenylmethoxycarbonyl/t-Bu chemistry. The peptide and the 1,2-diol were deprotected on the solid phase. Then, a periodic oxidation of the fully deprotected peptidyl-resin led to the simultaneous cleavage of the product from the solid support and to the generation of the alpha-oxo aldehyde moiety. The methodology allowed the distance between the alpha-oxo aldehyde and the peptide to be easily modulated. The C-terminal peptide alpha-oxo aldehydes synthesized in this study were found to be useful partners in hydrazone, thiazolidine, and oxime chemical ligations. PMID- 11397149 TI - Rapid homogeneous-phase Sonogashira coupling reactions using controlled microwave heating. AB - A microwave-enhanced, rapid and efficient homogeneous-phase version of the Sonogashira reaction is presented. It has been applied to the coupling of aryl iodides, bromides, triflates, and aryl chloride, as well as pyridine and thiophene derivatives with trimethylsilylacetylene. Excellent yields (80-95%) for substrates containing a large variety of substituents in different positions are obtained in 5-25 min. PMID- 11397150 TI - Synthesis and properties of a twistophane ion sensor: a new conjugated macrocyclic ligand for the spectroscopic detection of metal ions. AB - The synthesis of a structurally new type of conjugated macrocyclic ligand (1) is reported that comprises a dehydroannulene framework incorporating two 2,2' bipyridine units. Modeling studies showed the ligand to possess an unusual chirally twisted and relatively rigid architecture capable of binding metal ions in an enforced tetrahedral coordination geometry. The macrocycle was prepared in seven steps from (2-bromophenylethynyl)-trimethylsilane (2) and characterized by spectroscopic techniques. The pyridine H3 protons in the 1H NMR spectrum of 1 showed a marked temperature dependencey that may be related to conformational opening and closing motions of the macrocyclic ring. Ligand 1 was found to spectroscopically detect the presence of Co2+, Ni2+, Cu2+, and Zn2+ and, in particular, to function as a multiple readout sensor, giving different sequences of signal output depending upon the type of metal ion analyte with which the system was addressed. Macrocycle 1 also gave a highly characteristic and specific visual output response in the presence of Zn2+ consisting of a bright turquoise fluorescence and in this respect may find applications in the sensing of this biologically important metal ion. PMID- 11397151 TI - A beta-lactam-based stereoselective access to beta,gamma-dihydroxy alpha-amino acid-derived peptides with either alpha,beta-like or unlike configurations. AB - A concise access to alpha,beta-dihydroxy alpha-amino acid-derived N-carboxy anhydrides (NCAs) with either like or unlike relative configuration is described. The key steps of the synthetic route are the preparation of the nonracemic 4 alkenyl beta-lactams, through either Horner-type olefination of a common 4-formyl beta-lactam or the Corey-Winter alkene synthesis applied to 4-dihydroxyalkyl beta lactams, followed by the Sharpless AD reaction, and a subsequent ring expansion of the corresponding 4-substituted 3-hydroxy beta-lactams promoted by TEMPO. The opening of thus-prepared NCAs upon treatment with different O- and N nucleophiles, including alpha-amino esters which lead to peptides, has also been studied under various reaction conditions. PMID- 11397152 TI - A new synthesis of 2-azabicyclo[2.1.1]hexanes. AB - An efficient synthesis of the 2-azabicyclo[2.1.1]hexane ring system has been accomplished starting from cis-cyclobut-3-ene-1,2-dicarboxylic anhydride 7, which was prepared using a photochemical method. The key step of this new strategy involved a stereoselective electrophilic addition of phenylselenyl bromide to the double bond of cyclobutene dicarbamate 16 derived from 7. The subsequent ring closure of 17a in the presence of sodium hydride afforded the 2-azabicyclohexane compound 18 with a satisfying overall yield. Reductive removal of the phenylselenyl group and subsequent deprotection led rapidly to the amino derivative 4a functionalized on the carbon ring. Syntheses of the hydroxy and carboxylic derivatives 4b,c were then achieved from the intermediary disulfonamide 23. Displacement of the activated amino group by potassium acetate yielded hydroxy derivative 4b after three additional steps. Finally, oxidation of the alcohol function of 4b under Jones conditions followed by hydrogenolysis afforded the carboxylic derivative 4c, which is the first reported beta-isomer of 2,4-methanoproline 1. PMID- 11397153 TI - A general preparation of pyridines and pyridones via the annulation of ketones and esters. AB - A general preparation of pyridines 4a-f from stabilized ketones 3a-c and aryl ketones 3d-f is described. The annulation of stabilized esters 3g,h gives access to the corresponding 2-pyridones 4g,h. The annulation reactions proceed in fair to excellent yields (46-87%) with vinamidinium hexafluorophosphate salts 2a-d containing electron-withdrawing groups at the beta-position. The mechanism of the reaction was investigated by NMR and proceeds through the formation of a dienaminone intermediate. PMID- 11397154 TI - Stereospecific synthesis of the alpha- and beta-D-glucopyranosyl ureas. AB - A new, one-pot, two-stage procedure for the preparation of the alpha- and beta-D glucopyranosyl ureas has been developed. Oxidation of glucopyranosyl isocyanides provides glucopyranosyl isocyanates, which can be trapped in situ with amines to afford good yields of glucopyranosyl ureas. Application of this method establishes the successful synthesis of the hitherto unknown N,N'-di-alpha,alpha- and alpha,beta-D-glucopyranosyl ureas. PMID- 11397155 TI - Total synthesis and absolute configuration of minalemine A, a guanidine peptide from the marine tunicate Didemnum rodriguesi. AB - The total synthesis of the 3S,2S and 3R,2S diastereomers (1a and 1b) of minalemine A and the identification of the natural compound as the 3R,2S isomer is described. The key step in the synthesis is the preparation of the two enantiomers of the beta-amino diacid 3-(N-carboxymethyl)-aminodecanoic acid (Ncma), which were obtained by stereoselective alkylation with allyl bromide of two nonanoic acid imides bearing chiral oxazolidinones as chiral auxiliaries. Natural minalemine A shows identical 1H NMR and very similar 13C NMR spectra compared to the two synthetic diastereomers. Sufficient differences in their chromatographic behavior to allow conclusive identification were not found. However, the corresponding N-2-naphthoyl amides presented quite distinct circular dichroism spectra (CD), and these confirmed the 3R,2S configuration for the natural minalemines and the R configuration for the constituent beta-amino diacid, Ncma. PMID- 11397156 TI - Novel anionic annelation tactics for construction of fused heteroaromatic frameworks. 1. Synthesis of 4-substituted pyrazolo[3,4-c]quinolines, 9 substituted pyrazolo[3,4-c]quinolines, and 1,4-dihydrochromeno[4,3-c]pyrazoles. AB - 4-Substituted pyrazolo[4,3-c]quinolines 4a-i and 6a-b were prepared from pyrazole 3 whereas 9-substituted pyrazolo[3,4-c]quinolines 9a-d and 17 were prepared from pyrazole 13 utilizing anionic annelation techniques. 1,4-dihydrochromeno[4,3 c]pyrazoles 7a-c were accessed from pyrazole 3, extending the method for the synthesis of 4a-i. PMID- 11397157 TI - Studies on synthetic approaches to 1H- and 2H-indazolyl derivatives. AB - Synthetic approaches designed to provide 1H- and 2H-indazolyl derivatives of potential biological interest are reported. Special emphasis has been placed on the characterization of indazolylpyridinium products generated from reactions between indazole and 4-chloro-1-methylpyridinium iodide under various conditions. A stable mixture consisting of 3 parts of the 1H-isomer 9 to 1 part of the 2H isomer 10 was obtained at room temperature in the presence of the base 2,2,6,6 tetramethylpiperidine (TMP). The same reaction at 60 degrees C gave only the 1H isomer 9. At 100 degrees C in the absence of TMP only the 2H-isomer 10 was formed. The isomerization of 10 to 9 was found to proceed quantitatively at 60 degrees C but only in the presence of TMP. The effects of temperature and base on the course of these reactions are rationalized in terms of kinetic and thermodynamic parameters. PMID- 11397158 TI - Cal-B-catalyzed alkoxycarbonylation of a-ring stereoisomeric synthons of 1alpha,25-dihydroxyvitamin D3 and 1alpha,25-dihydroxy-19-nor-previtamin D3: a comparative study. first regioselective chemoenzymatic synthesis of 19-nor-A-ring carbonates. AB - A comparative study of alkoxycarbonylation processes of both 19-nor-A-ring and A ring stereoisomers of 1alpha,25-dihydroxyvitamin D3 analogues catalyzed by Candida antarctica lipase B (CAL-B) has been described. The presence of the methyl group in the A-ring at C-2, as in 3-6, has a determining role in the regioselectivity of the biocatalysis, mainly allowing the hydroxyl group at C-5 position to react. For the 19-nor-A-ring stereoisomers 7-10, which lack the C-2 methyl group, the configurations at C-3 and C-5 have a high influence in the selectivity exhibited by CAL-B. Thus, each couple of enantiomers showed opposing regioselectivities depending on the C-3 configuration. When C-3 possesses an (S) configuration, enzymatic alkoxycarbonylations took place at the C-5-(R) or C-5 (S) hydroxyl groups. However, if the chiral centers at C-3 are (R), CAL-B alkoxycarbonylated the C-3-(R) hydroxyl group independently of the configuration at C-5. The corresponding carbonates are useful A-ring precursors of 1alpha,25 dihydroxyvitamin D3 analogues, selectively modified at the C-1 or C-3 positions. In addition, an improved synthesis of cis A-ring synthons 5 and 6 is described using a Mitsunobu methodology. PMID- 11397159 TI - Solution and solid-support synthesis of a potential leishmaniasis carbohydrate vaccine. AB - The synthesis of a potential carbohydrate vaccine for the parasitic disease leishmaniasis is described. New solution- and solid-phase synthetic strategies were explored for the assembly of a unique tetrasaccharide antigen found on the Leishmania lipophosphoglycan. An initial solution-phase synthesis relied on thioglycosides as building blocks and the establishment of the central disaccharide from lactal via an oxidation-reduction sequence. A second approach was completed both in solution and on solid support. The solid-phase synthesis relied on assembly from monosaccharide units and was used to evaluate different glycosylating agents in the efficient installation of the galactose beta-(1-->4) mannoside. Glycosyl phosphates proved most successful in this endeavor. This first solid-phase synthesis of the Leishmania cap provided rapid access to the tetrasaccharide in 18% overall yield while requiring only a single purification step. The synthetic cap tetrasaccharide was conjugated to the immunostimulator Pam3Cys to create fully synthetic carbohydrate vaccine 1 and to the carrier protein KLH to form semisynthetic vaccine 2. Currently, both constructs have entered initial immunological experiments in mice targeted at the development of a vaccine against the parasitic disease leishmaniasis. PMID- 11397160 TI - Effects of aromatic thiols on thiol-disulfide interchange reactions that occur during protein folding. AB - The folding of disulfide containing proteins from denatured protein to native protein involves numerous thiol-disulfide interchange reactions. Many of these reactions include a redox buffer, which is a mixture of a thiol (RSH) and the corresponding disulfide (RSSR). The relationship between the structure of RSH and its efficacy in folding proteins in vitro has been investigated only to a limited extent. Reported herein are the effects of aliphatic and especially aromatic thiols on reactions that occur during protein folding. Aromatic thiols may be particularly efficacious as their thiol pK(a) values and reactivities match those of the in vivo catalyst, protein disulfide isomerase (PDI). This investigation correlates the thiol pK(a) values of aromatic thiols with their reactivities toward small molecule disulfides and the protein insulin. The thiol pK(a) values of nine para-substituted aromatic thiols were measured; a Hammett plot constructed using sigma(p-) values yielded rho = -1.6 +/- 0.1. The reactivities of aromatic and aliphatic thiols with 2-pyridyldithioethanol (2-PDE), a small molecule disulfide, were determined. A plot of reactivity versus pK(a) of the aromatic thiols had a slope (beta) of 0.9. The ability of these thiols to reduce (unfold) the protein insulin correlates strongly with their ability to reduce 2 PDE. Since the reduction of protein disulfides occurs during protein folding to remove mismatched disulfides, aromatic thiols with high pK(a) values are expected to increase the rate not only of protein unfolding but protein folding as well. PMID- 11397161 TI - De novo synthesis of a methylene-bridged Neu5Ac-alpha-(2,3)-gal C-disaccharide. AB - A general strategy toward the synthesis of C-ketosides of N-acetylneuraminic acid (Neu5Ac) has been developed and successfully applied to the synthesis of methylene-bridged Neu5Ac-alpha-(2,3)-Gal C-disaccharide 2. The key strategic element of this novel approach is a stereoselective, 6-exo-trig selective, electrophilic cyclization of the appropriate open chain precursor 4 by means of phenylselenyl triflate. The open chain precursor was formed by the addition of lithiated iodide 18 accessible from D-galactose to open chain aldehyde 5a obtained from D-glucono-delta-lactone by chain elongation. Subsequent C1 incorporation using Tebbe-reagent, formation of a cyclic carbonate, and deprotection of the two isopropylidene ketals afforded tetrol 4 which, upon treatment with phenylselenyl triflate, was stereoselectively cyclized in a 6-exo trig selective manner. A selena-Pummerer rearrangement, oxidation, and esterification readily led to methyl ester 37 which, after deacetylation, could be regioselectively tetrabenzoylated with benzoyl cyanide. Triflate activation of the axial hydroxyl group in 40 and nucleophilic displacement by azide ion with inversion of configuration afforded azide 41, which was reduced with hydrogen and Pearlman's catalyst. Concomitant removal of the benzyl ethers and subsequent saponification of all ester moieties successfully completed the de novo synthesis of the desired methylene bridged Neu5Ac-alpha-(2,3)-Gal C-disaccharide 2. PMID- 11397162 TI - Fluorous Boc ((F)Boc) carbamates: new amine protecting groups for use in fluorous synthesis. AB - The first fluorous variants of the Boc (tert-butyloxycarbonyl) group have been prepared and tested for their suitability as nitrogen protecting groups. A group with two fluorous chains and an ethylene spacer, (RfCH2CH2)2(CH3)COC(O)-, was readily attached to a representative amine but was difficult to cleave. In contrast, groups with two fluorous chains and a propylene spacer, (RfCH2CH2CH2)2(CH3)COC(O)-, or one fluorous chain and an ethylene spacer, (RfCH2CH2)(CH3)2COC(O)-, were readily formed and cleaved. The fluorous alcohol component of the (F)Boc group can be removed by evaporation and can be recovered and reused. The utility of the new (F)Boc group (C8F17CH2CH2)(CH3)2COC(O)- was demonstrated in 16 and 96 compound library synthesis exercises. Separations can be achieved either by manual, parallel fluorous solid-phase extraction, or automated, serial fluorous chromatography. The results provide additional confirmation of the value of "light" fluorous synthesis techniques, and the new fluorous Boc groups expand the applicability of fluorous synthesis techniques to many classes of nitrogen-containing organic compounds. PMID- 11397163 TI - A simple and versatile one-pot synthesis of meso-substituted trans-A2B-corroles. AB - We have developed a new methodology that affords regioisomerically pure trans-A2B corroles. The corrole formation reaction involves the acid-catalyzed condensation of a dipyrromethane and an aldehyde followed by oxidation with DDQ. Optimal conditions for the condensation were identified after examining various reaction parameters (solvent, acid, concentration, time). The conditions identified (CH2Cl2, [DPM] = 33 mM, [aldehyde] = 17 mM, [TFA] = 1.3 mM (for sterically hindered DPMs) or [TFA] = 0.26 mM (for sterically unhindered DPMs), 5 h, room temperature) resulted in the formation of corroles in 3-25% yield without detectable scrambling. The synthesis is compatible with diverse functionalities: ester, nitrile, ether, fluoro, hydroxy, iodo, nitro, thioacetate, methylsulfoxy. In total 21 corroles of type trans-A2B were prepared. Three exemplary corrole syntheses were successfully carried out at 8 mmol scale. Corroles 23, 30, and 41 (160-600 mg) were obtained in essentially the same yield as in small scale experiments. PMID- 11397164 TI - Synthesis of (+)-1-epiaustraline. AB - A highly efficient total synthesis of (+)-1-epiaustraline ((+)-1), a tetrahydroxypyrrolizidine alkaloid of the alexine/australine subclass, is described. The key step is a tandem intramolecular [4 + 2]/intermolecular [3 + 2] nitroalkene cycloaddition involving dienylsilyloxy nitroalkene 3 and chiral vinyl ether 4, which establishes four of the five stereocenters present. The final center was installed by a diastereoselective dihydroxylation. Hydrogenolytic unmasking of the nitroso acetal tosylate 17 containing the silyl ether linkage was thwarted by a slow alkylation and an undesired Peterson-type elimination. Prior removal of the silicon moiety by Tamao-Fleming oxidation proceeded in excellent yield and provided a substrate suitable for hydrogenolysis and deprotection. The complete synthesis required only 10 steps to deliver the (+)-1 epiaustraline in 7.0% overall yield. PMID- 11397165 TI - Modern valence-bond description of chemical reaction mechanisms: the 1,3-dipolar addition of diazomethane to ethene. AB - The electronic mechanism for the gas-phase concerted 1,3-dipolar cycloaddition of diazomethane (CH2N2) to ethene (C2H4) is described through spin-coupled (SC) calculations at a sequence of geometries along the intrinsic reaction coordinate obtained at the MP2/6-31G(d) level of theory. It is shown that the bonding rearrangements occurring during the course of this reaction follow a heterolytic pattern, characterized by the movement of three well-identifiable orbital pairs, which are initially responsible for the pi bond in ethene and the C-N pi bond and one of the N-N pi bonds in diazomethane and are retained throughout the entire reaction path from reactants to product. Taken together with our previous SC study of the electronic mechanism of the 1,3-dipolar cycloaddition of fulminic acid (HCNO) to ethyne (C2H2) (Theor. Chim. Acc. 1998, 100, 222), the results of the present work suggest strongly that most gas-phase concerted 1,3-dipolar cycloaddition reactions can be expected to follow a heterolytic mechanism of this type, which does not involve an aromatic transition state. The more conventional aspects of the gas-phase concerted 1,3-dipolar cycloaddition of diazomethane to ethene, including optimized transition structure geometry, electronic activation energy, activation barrier corrected for zero-point energies, standard enthalpy, entropy and Gibbs free energy of activation, have been calculated at the HF/6 31G(d), B3LYP/6-31G(d), MP2/6-31G(d), MP2/6-31G(d,p), QCISD/6-31G(d) and CCD/6 31G(d) levels of theory. We also report the CCD/6-311++G(2d, 2p)//CCD/6-31G(d), MP4(SDTQ)/6-311++G(2d,2p)//CCD/6-31G(d) and CCSD(T)/6-311++G(2d, 2p)//CCD/6 31G(d) electronic activation energies. PMID- 11397166 TI - Catalytic Asymmetric Vinylogous Mukaiyama-aldol (CAVM) reactions: the enolate activation. AB - The Catalytic Asymmetric Vinylogous Mukaiyama (CAVM) reactions of various aldehydes with dienolate 1 using different enolate activations (CuF*(S)-TolBinap, t-BuOCu*(S)-Tol-Binap, and various chiral nonracemic ammonium fluorides derived from cinchona alkaloids) are described. These reactions proved to be highly regioselective leading exclusively to the alpha-aldol products in good yields and poor to good enantioselectivities. PMID- 11397167 TI - Syntheses and structures of isomeric [6.6]- and [8.8]cyclophanes with 1,4 dioxabut-2-yne and 1,6-dioxahexa-2,4-diyne bridges. AB - All three isomers (ortho, meta, and para) of [8.8]cyclophane bearing 1,6 dioxahexa-2,4-diyne bridges have been synthesized and structually characterized by single-crystal X-ray crystallography to determine the conformation of the cyclophanes and their cavity dimensions. The three isomeric [6.6]cyclophanes bearing 1,4-dioxabut-2-yne bridges have also been synthesized from but-2-yne-1,4 diol ditosylate and the isomeric dihydroxybenzenes. The [6.6]orthocyclophane has been structurally characterized by single-crystal X-ray crystallography. The energy-minimized structures from the semiempirical AM1 calculations of these cyclophanes compare very well with the structures obtained by X-ray crystallography. PMID- 11397168 TI - Evaluation of exo-endo ratios in the halolactonization of omega-unsaturated acids. AB - The reaction of 2-(omega-alkenyl)benzoic acids with bis(collidine)iodine and bis(collidine)bromine hexafluorophosphate was examined. Except with 2-but-3 enylbenzoic acid, for which only the exo lactone was obtained, for the other acids a mixture of exo-endo lactones was always obtained. The proportion of endo lactone was important for the acid chain length of 11 carbons (formation of a 12 membered ring endo lactone) and for the acid chain lengths higher than 14 carbons. The formation of the endo lactones was explained, on the base of molecular calculations, by competition between electronic and steric effects. These latter were developed by transannular interactions (for the acid chain lengths 8-11) and/or the conformations adopted by the chains (for the acid chain lengths > or = 14,) which disfavored the formation of the exo lactones. The larger proportion of endo lactones observed with the bromo reagent compared to the iodo reagent seemed due to electronic factors. PMID- 11397169 TI - Synthesis of primary aromatic amides by aminocarbonylation of aryl halides using formamide as an ammonia synthon. AB - Primary aromatic amides were prepared by a palladium-catalyzed aminocarbonylation reaction of aryl halides in high yields (70-90%) using formamide as the amine source. The reactions require a palladium catalyst in combination with a nucleophilic Lewis base such as imidazole or 4-(dimethylamino)pyridine (DMAP). Aryl, heteroaryl, and vinyl bromides and chlorides were converted to the primary amides under mild conditions (5 bar, 120 degrees C) using 1 mol % of a palladium phosphine complex. Best results were obtained in dioxane using triphenylphosphine as the ligand and DMAP as the base. For activated aryl bromides, a phosphine-to palladium ratio of 2:1 was sufficient, but less reactive aryl bromides or aryl chlorides required ligand-to-palladium ratios up to 8:1 in order to stabilize the catalyst and achieve full conversion. The influence of catalyst, base, solvent, pressure, and temperature was studied in detail. The mechanism of the reaction could be clarified by isolating and identifying the reaction intermediates. In addition, methylamides and dimethylamides were prepared by the same method using N-methylformamide and N,N-dimethylformamide as the amine source. PMID- 11397170 TI - Theoretical Study of [2 + 1] cycloaddition of CO and CS to acetylenes forming cyclopropenones and cyclopropenethiones. AB - The [2 + 1] cycloadditions of carbon monoxide and carbon monothioxide CX (X = O, S) to acetylenes (R1C triple bond CR2 with R1 = H, OH and R2 = CH3, OH, NH2, C6H5) have been studied at the B3LYP/6-311G(d,p) level. It has been shown that the reaction proceeds in two steps forming first an intermediate having the properties of both a carbene and a zwitterion followed by a ring closure leading to cyclopropenones or cyclopropenethiones. The solvent effect does not play an important role in the course of the cycloaddition. The estimation of the first vertical excitation energies by CIS and TD-B3LYP methods shows that the reactions likely take place in the ground state rather than in an excited state. All the studied cyclopropenones and cyclopropenethiones are aromatic as shown by their NICS values and confirmed by calculated and experimental NMR chemical shifts. Different reactivity criteria including HOMO coefficient, local softness, hardness, polarizability, and NICS are used to predict the site selectivity in all studied cases, and the NICS criterion seems to yield the best results among them. PMID- 11397171 TI - Novel rearrangements of sesquiterpenoid panasinsane derivatives under acidic conditions. AB - The sesquiterpenoid panasinsane derivatives 11 and 14-16 have been prepared from caryophyllene oxide (7). The novel rearrangement reactions of compounds 11 and 14 under TCNE-catalyzed solvolysis conditions and the reactions of compounds 15 and 16 under superacid conditions (HSO3F/Et2O, -63 degrees C) have been investigated. The ginsenol derivative 17 is obtained from compounds 11 and 14 under TCNE catalyzed conditions. The rearrangement of compounds 15 and 16 under superacid conditions leads to the novel sesquiterpene derivatives (1S,4S,7S,10S,11S) 3,3,10,11-tetramethyltricyclo[5.3.1.0(4,10)]undecan-1,11-yl sulfate (19) and (1S,4S,5S,8S)-2,2,4,8-tetramethyl tricyclo[3.3.2.1(4,8)]undecan-11-one (20). The influence of the secondary hydroxyl group at C-5 of the panasinsane derivatives on the course of these rearrangements is discussed. PMID- 11397172 TI - Selective halogen-magnesium exchange reaction via organomagnesium ate complex. AB - Halogen-magnesium exchange of various aryl halides is achieved with a magnesium ate complex at low temperatures. Tributylmagnesate (nBu3MgLi) induces facile iodine-magnesium exchange at -78 degrees C. Dibutylisopropylmagnesate (iPr(n)Bu2MgLi) is more reactive than nBu3MgLi, and this reagent accomplishes selective bromine-magnesium exchange at -78 degrees C. This procedure is utilized for the preparation of various polyfunctionalized arylmagnesium species. The exchange of alkenyl halides using this method proceeds with retention of configuration of the double bond. PMID- 11397173 TI - Aqueous DMF-potassium carbonate as a substitute for thallium and silver additives in the palladium-catalyzed conversion of aryl bromides to acetyl arenes. AB - Highly selective palladium-catalyzed internal alpha-arylations of alkyl vinyl ethers with aryl and heteroaryl bromides were conveniently conducted in aqueous DMF with potassium carbonate as base and with DPPP as bidentate ligand. The corresponding acetyl arene products were, after hydrolysis, isolated in good to excellent yields. This Heck reaction procedure does not require toxic thallium or expensive silver salt additives, is promoted by water, and is suggested to proceed via charged organopalladium intermediates. Single-mode microwave irradiation was utilized in one example to shorten the reaction time. PMID- 11397174 TI - A diastereoselective intramolecular hydroamination approach to the syntheses of (+)-, (+/-)-, and (-)-pinidinol. AB - A diastereoselective, lanthanocene-catalyzed, intramolecular hydroamination reaction was applied to the preparation of 2,6-disubstituted piperidines. Various metal/ligand arrays in the catalysts were examined using a model substrate to allow optimization of the diastereoselectivity. It was determined that the relationship between metal size and ligand bulk plays an integral role in the transformation. The complex Cp2NdCH(TMS)2 converted 2-substituted 8-nonen-4 amines to 2,6-disubsituted piperidines with greater than 100:1 selectivity for the formation of the cis isomer. A short synthesis of pinidinol, an alkaloid isolated from various pine and spruce species, was then carried out to exploit this stereoselective reaction. PMID- 11397175 TI - Acetyltrimethylsilane, trifluoromethyltrimethylsilane, and prenyl esters: a three component system for the synthesis of gem-difluoroanalogues of monoterpenes. AB - The preparation of 3,3-difluoro-6-methylhept-5-en-2-one 1, a key intermediate for the synthesis of 4,4-difluoroterpenes, and applications in linalool and geraniol series are described. The process involves 1,1-difluoro-2 trimethylsilyoxypropene, an enol silyl ether prepared from acetyltrimethylsilane and trifluoromethyltrimethylsilane, and its reaction in situ with prenyl benzoate, under catalysis by trimethylsilyl trifluoromethanesulfonate. Optimized conditions leading to either the desired enol silyl ether or the unprecedented methyl(trifluoromethyl)trimethylsilyl carbinol 4 have been achieved. The prenylation of the enol silyl ether gives a 9/1 mixture of regioisomers, in favor of the expected ketone 1. Treatment of 1 with vinylmagnesium bromide leads to (+/ )-4,4-difluorolinalool 7. Reaction with the lithium enolate of ethyl diethylphosphonoacetate, and then LAH reduction, converts 1 to 4,4 difluorogeraniol 11, with complete stereoselectivity. PMID- 11397176 TI - A highly stereoselective approach to the synthesis of functionalized pyran derivatives by Lewis acid assisted ketal reduction and allylation. AB - Reduction of bicyclic ketal 1 gave functionalized pyran derivatives 7a or 7b in a highly stereoselective manner, depending upon the reduction conditions utilized. For example, treatment of ketal 1 with TiCl4/Et3SiH produced exclusively diol 7b with the 2,5-syn relationship in good yield. Alternatively, reduction of ketal 1 by DIBALH gave 2,5-anti-diol 7a stereoselectively. Alane reductions of ketal 1 were highly stereoselective also; however, the syn/anti selectivity observed was strongly dependent on the ratio of reagents employed for in situ generation of the alane. Lewis acid catalyzed allylation of ketal 1 gave pyran 10 in a stereospecific alkylation reaction. PMID- 11397177 TI - Nonacid nitration of benzenedicarboxylic and naphthalenecarboxylic acid esters. AB - When treated with nitrogen dioxide in the presence of ozone and a catalytic amount of iron(III) chloride in inert organic solvent at -10 to +5 degrees C, benzenedicarboxylic acid diesters 1, 4, and 6 underwent smooth nitration to give the corresponding mononitro derivatives 2/3, 5, and 7, respectively, in good yield (kyodai nitration). Naphthalenecarboxylic acid esters 8 and 11 and naphthalene-1,8-dicarboxylic acid diester 16 were similarly nitrated in the absence of catalyst to give the expected nitro compounds 9/10, 12-15, and 17-22, respectively. Different from conventional nitration based on the combined use of concentrated nitric and sulfuric acids, no hydrolytic cleavage of the ester function was observed under these conditions. The isomer distribution has been determined for the nitration of naphthalenecarboxylic acid esters 8, 11, and 16, and spectral data were collected for less common nitro derivatives. A unique changeover of the orientation mode observed in the kyodai nitration of diester 16, from the initial exclusive meta to the final meta/para, has been discussed in terms of the competition between the electrophilic substitution process involving the nitronium ion (NO2+) and the addition-elimination sequence involving the nitrogen trioxide radical (*NO3). PMID- 11397178 TI - Synthesis of (+/-)-bakkenolide-A and its C-7, C-10, and C-7,10 epimers by means of an intramolecular Diels-Alder reaction. AB - (+/-)-bakkenolide-A (1) was prepared in five steps from ethyl 4 benzyloxyacetoacetate by sequential alkylations with tiglyl bromide and (Z)-5 bromo-1,3-pentadiene, followed by an intramolecular Diels-Alder reaction of (E,Z) triene 25b as the key step. The hydrindane cycloadduct 28 was subjected to hydrogenation and spontaneous or acid-catalyzed lactonization, followed by a Witttig reaction to introduce the exocyclic methylene group of 1. The known 7 epibakkenolide-A (2) and novel 10-epi- and 7,10-diepibakkenolide-A (3 and 4, respectively) stereoisomers were obtained as minor byproducts. When (E)-5-bromo 1,3-pentadiene was used instead of the Z-isomer, the 10-epi- and 7,10 diepibakkenolides were the major products. In both cases exo cyclization was preferred over endo. An alternative approach was based on a similar intramolecular Diels-Alder cycloaddition, using dimethyl malonate instead of ethyl 4-benzyloxyacetoacetate as the starting material for the double alkylation preceding the cycloaddition step. The cycloadduct was then converted into the corresponding alpha-phenylseleno propargyl esters 16 or 22. However, attempted formation of the spiro center by a radical cyclization resulted chiefly in reductive deselenization. PMID- 11397179 TI - On the interactivity of complex synthesis and tumor pharmacology in the drug discovery process: total synthesis and comparative in vivo evaluations of the 15 aza epothilones. AB - The total syntheses of 12,13,15-desoxy-15(S)-aza-epothilone B (aza-dEpoB; dEpoB lactam) and 12,13,15-desoxy-15(R)-aza-epothilone B (15-epi-aza-dEpoB; 15-epi dEpoB-lactam) have been accomplished via a highly convergent strategy. We have also successfully oxidized 12,13,15-desoxy-15(S)-aza-epothilone B to aza epothilone B (aza-EpoB; EpoB-lactam). Aza-epothilone B has been advanced to phase I clinical trials by the Bristol-Myers Squibb group. Our synthesis is efficient and was amenable to the production of significant quantities of these lactams. Using our fully synthetically derived lactams, in vitro and in vivo studies were conducted in comparison with advanced clinical candidates, 12,13-desoxyepothilone B and 12,13-desoxyepothilone F, also derived by total synthesis. PMID- 11397180 TI - Enantiospecific total synthesis of (-)-4-thiocyanatoneopupukeanane. AB - Enantiospecific synthesis of the natural enantiomer of the marine sesquiterpene ( )-4-thiocyanatoneopupukeanane (6) is described. The bicyclo[2.2.2]octanecarboxylate 14, obtained from (R)-carvone via Michael-Michael reaction, was transformed into neopupukeananedione 12 by employing rhodium acetate catalyzed intramolecular C-H insertion of the diazo ketones 16 or 19 as the key reaction. Regioselective deoxygenation of the C-2 ketone transformed the dione 12 into neopupukean-4-one 10. Alternately, the keto ester 18 was also transformed into neopupukean-4-one 10 via regioselective deoxygenation of the ketone in 18 followed by intramolecular rhodium carbenoid C-H insertion of the diazo ketone 31. Finally, neopupukean-4-one 10 was transformed into (-)-4 thiocyanatoneopupukeanane 6 via the alcohol 32 and the mesylate 33. PMID- 11397181 TI - 2,6-Dichloro-9-thiabicyclo[3.3.1]nonane: a privileged, bivalent scaffold for the display of nucleophilic components. AB - The title compound, the condensation product of sulfur dichloride and 1,5 cyclooctadiene, is a reliable acceptor of a wide variety of heteroatom nucleophiles, sometimes in reversible fashion. Optical resolution of the core structure has been achieved and preserved in succeeding transformations. The high reactivity and reliable stereochemical control afforded by this system illustrates the power of neighboring-group participation by the sulfur center. PMID- 11397182 TI - Reductive electrochemistry of spiromethanofullerenes. AB - The synthesis and electrochemical properties of spiromethanofullerenes 1-6 are described. The syntheses were achieved via the diazomethane route, and the regioisomeric distribution of bis-adducts 3 was determined by (1)H NMR, UV-vis, and HPLC analysis. The electrochemistry of these compounds in dichloromethane exhibits, besides several reversible reductions, some irreversible waves. Reductive electrolysis of 1 and 3-5 in dichloromethane leads to the removal of the addend and thus to the formation of C60. Reductive electrolysis of 1, 2 and 6 in dichloromethane leads to a different reaction, which we tentatively assign to a reaction with the solvent to form fullerene-CH2Cl products. Electrolyses in the less reactive solvent THF lead to adduct removal for 1 and 3-5. Interestingly, significant formation of bis-adducts from the corresponding monoadducts was observed for 4 and 5. These electrochemically induced reactions, namely addend removal, reaction with the solvent, and addend transfer are influenced by the structure of the addend and by the choice of solvent. PMID- 11397183 TI - Effect of beta-cyclodextrin on the hydrolysis of trifluoroacetate esters. AB - The hydrolysis of p-F, p-Cl, and m-Cl phenyl trifluoracetates was studied in the presence of beta-cyclodextrin (beta-CD). The reactions are inhibited by beta-CD at pH 6 while they are catalyzed in alkaline solution. MM3 calculations reproduce some of the experimental results. The substrates form inclusion complexes with beta-CD which are of similar stability as those of the corresponding acetates; however, the association of the transition state is less favorable in these reactions than in those of the acetates, and consequently less stronger catalysis is observed. PMID- 11397185 TI - Rearrangements in the reduction of 3-iodobicyclo[1.1.1]pentyl azide with lithium aluminum hydride: mechanistic evidence of intermediates. PMID- 11397184 TI - Investigation of an unexpected addition reaction that occurs when 2,3:4,5-di-O isopropylidene-D-ribose diethyl dithioacetal is treated with mercuric oxide and mercuric chloride. PMID- 11397186 TI - Chemoselective cyclization of aminonicotinic acid derivatives to 1,8-naphthyridin 2-ones via a potential intramolecular azadiene-ketene electrocyclization reaction. PMID- 11397187 TI - Unexpected formation of a spiro acridine and fused ring system from the reaction between an N-acridinylmethyl-substituted thiourea and bromoacetonitrile under basic conditions. PMID- 11397188 TI - Crown-linked porphyrin systems. PMID- 11397189 TI - A novel synthesis of 2-aminopyrroles using a three-component reaction. PMID- 11397190 TI - Selective deprotection of N-Boc-protected tert-butyl ester amino acids by the CeCl3*7H2O-NaI system in acetonitrile. PMID- 11397191 TI - Iridium(I)-catalyzed cycloisomerization of enynes. PMID- 11397192 TI - A novel and facile racemization of chiral 1,1'-biaryl-2,2'-dicarboxylic acids. PMID- 11397193 TI - Electrochemical activation of dimethyl disulfide for electrophilic aromatic substitution. PMID- 11397194 TI - Conformational studies by dynamic NMR. 81. cogwheeling circuit for the enantiomerization of the propeller antipodes of 2,2',6,6'-tetramethyldiphenyl sulfide. PMID- 11397195 TI - Practical synthesis of myrcene derivatives possessing oxidized methyl groups. PMID- 11397196 TI - BF3*OET2-mediated rearrangement of cyclopropyl carbinols: a concise route to polycyclic cyclobutanes. PMID- 11397197 TI - Highly stereoselective synthesis of trisubstituted alpha,beta-unsaturated sulfoxides by Heck reaction. PMID- 11397198 TI - Ring annulation with tetrahydroisoquinoline-derived enaminones: highly convergent routes to functionalized pyrrolo[2,1-a]- and indolo[2,1-a]isoquinolines. PMID- 11397202 TI - Modulation of proteolytic activity associated with persistent corneal ulcers in dogs. AB - Canines affected with corneal lesions demonstrate increased proteolytic activity at the affected site. Canines that develop persistent corneal ulcers and maintain elevated levels of proteolytic activity respond to protease inhibitor therapy with polysulpated glycosaminoglycan, PSGAG. In this study, the proteolytic activity expressed in lacrimal fluid of canines was evaluated as normal (1.15 U mL-1) or healed (1.19 U mL-1). Six of the 26 dogs affected with persistent corneal ulcers, however, had a protease level consistent with the unaffected control animals. These dogs did not appear to respond to PSGAG therapy. Two pathophysiologies for persistent corneal ulcers are indicated. Identifying which mode is active could help determine the therapeutic treatment needed. PMID- 11397203 TI - An outbreak of cataract with lens rupture and nuclear extrusion in wolf-fish (Anarhicas spp.). AB - Eight spotted (Anarhicas minor Olafsen) and five common wolf-fish (Anarhicas Lupus L), developed cataracts shortly after an episode of increased water temperature and decreased salinity 5 years prior to examination. On clinical examination, the cataracts were mostly bilateral and complete, and a majority of the lenses were lobulated. Inflammatory reaction was, apart from one eye with severe inflammation, limited to iris atrophy. Of the 14 eyes collected for pathomorphological examination, eight had lens rupture with extrusion of the nucleus to the posterior chamber, two showed partly dislocated nuclei with posterior protrusion and two lenses were morgagnian. A multilayered squamous epithelium with abundant desmosomes had developed on the surface of seven of the extruded nuclei. The main cause of the cataracts was considered to be the rapid decrease in water salinity, causing osmotic changes within the eyes with secondary swelling of lens fibers and rupture of the lens capsules. PMID- 11397204 TI - Idiopathic Horner's syndrome in collie dogs. AB - Seven cases of idiopathic Horner's Syndrome in the Collie are described. Five males and two females presented with unilateral miosis, ptosis of the upper eyelid, enophthalmos and protrusion of the third eyelid. Thorough examination, pharmacological testing with phenylephrine, complete blood counts and radiography of the tympanic bullae and thorax were performed. The etiology was not identified in any of the cases. Clinical signs improved with pharmacologic testing within 20 40 min. In five dogs, total resolution of clinical signs was observed at 4 and 16 weeks after their initial appearance. Pharmacological testing suggested that the deficit could be at the preganglionic neuron. PMID- 11397205 TI - Tonometry in three herbivorous wildlife species. AB - Tonometry was performed to estimate intraocular pressure (IOP) in 12 Nubian ibexes (Capra ibex nubiana), 10 Grant zebras (Equus burchelli ) and five Arabian oryxes (Oryx leucoryx), using both applanation (Tono-Pen) and/or indentation (Schiotz) tonometers. Animals were anesthetized with a mixture of etorphine hydrochloride and acepromazine maleate. Mean (+/- SD) IOP in the ibex was 17.95 +/- 4.78 mmHg (24 eyes, indentation tonometry). In the zebra, indentation tonometry (20 eyes) yielded a mean IOP of 25.30 +/- 3.06 mmHg, and applanation tonometry (six eyes) yielded a mean IOP of 29.47 +/- 3.43 mmHg. In the oryx, indentation tonometry (five eyes) yielded a mean IOP of 22.68 +/- 8.15 mmHg, and applanation tonometry (10 eyes) yielded a mean IOP of 11.76 +/- 3.43 mmHg. There were no significant effects of gender, age, weight, side or reading number on the IOP measured in any of the three species. No significant differences were found between the IOP of the three species, nor between the readings of the two instruments, although some of the P-values were close to the significance level. PMID- 11397206 TI - A matched observational study of survival in cats with enucleation due to diffuse iris melanoma. AB - Although a small number of cases of feline diffuse iris melanoma have been documented to metastasize, the prognosis is not known. In this matched observational study, the survival time of 34 cats with enucleation due to histologically confirmed diffuse iris melanoma was recorded. These results are compared to the survival times of 83 age-matched control cats. Affected cats had enucleation between 2 and 10 years prior to the study. One group of control cats with eye disease had enucleation for either lymphoplasmacytic uveitis (27 cases), ocular trauma (seven cases), or endophthalmitis (four cases). In these control cats, enucleations were performed between 2 and 10 years prior to this study. Forty-five additional control cats presented for vaccination between 2 and 10 years prior to the study. The extent of diffuse iris melanoma at the time of enucleation in affected cats was graded according to the extent of involvement of ocular tissues and the invasiveness of the tumor. Affected cats have a significantly decreased survival compared with control cats and cats with extensive tumors at the time of enucleation have the lowest survival rates. Cats with tumors confined to the iris survive at the same rate as controls. These results suggests that early enucleation is important to avoid premature death, presumed to be due to cancer metastasis in cats with diffuse iris melanoma. PMID- 11397207 TI - Anterior chamber to frontal sinus shunt for the diversion of aqueous humor: a pilot study in four normal dogs. AB - Silicone tubing was used to divert aqueous humor from the anterior chamber of the right eye to the rostral compartment of the right frontal sinus in four clinically normal mixed-breed dogs. Biomicroscopic examination, and pneumoapplanation tonometry and tonography, completed for up to 18 weeks postoperatively, confirmed gonioimplant function in all four cases. The dogs were euthanized at 6, 8, 16 and 18 weeks postoperatively. Gonioimplant patency was further confirmed by postmortem examination of the globes, implants and frontal sinuses. Gross and light microscopic examinations revealed iridal attachments to the implant (n = 4), mild anterior uveitis (n = 3), anterior subcapsular cataracts (n = 4), and focal corneal (n = 3) and scleral (n = 3) scarring in the operated globes. Light microscopic examination of frontal sinus specimens revealed mild lymphocytic proliferation and fibrosis immediately adjacent to the implant entrance site. There were no bacteria detected on aerobic or anaerobic cultures of the frontal sinuses or light microscopic examination of the globes or frontal sinuses. Results indicate that the frontal sinus shunting of aqueous humor is a safe and effective means of extraorbital aqueous diversion with potential applicability in the management of glaucoma. PMID- 11397208 TI - Multiple ciliary body cysts and secondary glaucoma in the Great Dane: a report of nine cases. AB - Between 1989 and 1995 nine Great Danes were presented to the small animal clinic of the University of Zurich Teaching Hospital with glaucoma and multiple cysts in the anterior and posterior chamber. In four of the nine dogs cysts were present in both eyes; however, bilateral glaucoma was seen in one case only. Mean intraocular pressure (IOP) at initial presentation was 42 mmHg measured by applanation tonometry. With the exception of one dog, all animals were treated medically for a minimum of 13 days. Two animals were subsequently lost to follow up. Two dogs underwent evisceration with implantation of a silicone prosthesis. The glaucomatous globe was enucleated in three dogs. One owner declined surgery and the dog remained buphthalmic with a poorly controlled IOP. One animal remained visual after trans-scleral diode laser cyclophotocoagulation. Histopathology of the three enucleated globes showed multiple cysts originating from the ciliary body epithelium. The mechanism of IOP elevation is probably by anterior displacement of the iris with narrowing of the angle and collapse of the ciliary cleft. A delicate pre-iridal fibrovascular membrane was also seen in each case, which could have also contributed to the elevation of IOP. The high odds ratio of 2.23 (CI 95% 1.14, 3.99) for glaucoma and 37.01 (CI 95% 16.42, 77.81) for ciliary body cysts suggests they are both inherited in this breed. Pedigree analysis of the affected dogs failed to definitely reveal the mode of inheritance. PMID- 11397209 TI - Uveodermatological syndrome (Vogt-Koyanagi-Harada-like syndrome) with generalized depigmentation in a Dachshund. AB - A 3-year-old, female, black and tan Dachshund was referred with visual impairment, bilateral anterior and posterior uveitis, poliosis, and generalized dermal depigmentation. Complete blood counts and biochemical parameters, including T3 and T4, were normal. The skin biopsy showed lichenoid dermatoses with dermal infiltration of histiocytes and lymphocytes suggesting uveodermatological syndrome. Medical treatment was initiated with oral prednisone and azathioprine, and topical prednisolone and atropine. The clinical signs improved, vision was retained, and the skin began to repigment 2 months following the initiation of therapy. PMID- 11397210 TI - 'D-shaped' and 'reverse-D-shaped' pupil in a cat with lymphosarcoma. AB - A 7-year-old female spayed European shorthair cat was presented because of visual reduction and a green reflex in the right eye. Examination revealed a 'D-shaped' pupil in the right eye and a 'reverse-D-shaped' pupil in the left eye. Both eyes showed fibrinous anterior uveitis with exudates in the anterior chamber and precipitates on the posterior surface of the cornea. In both eyes a chorioretinitis was diagnosed which caused partial retinal detachment and peripapillary oedema in the left eye. Histopathology revealed intestinal and ocular lymphosarcoma. The aberrant pupil shapes which persisted after death were due to infiltration of the iris stroma and were not caused neurologically. The observation that in both eyes only the medial parts of the iris were affected could not be explained. PMID- 11397211 TI - Effects of intramuscular sedative and opioid combinations on tear production in dogs. AB - The effect of commonly used sedation protocols on tear production rate was evaluated in dogs. Schirmer I tear tests were examined before and after intramuscular injection of acepromazine and oxymorphone (ACE + OXY; n = 7), diazepam and butorphanol (DIA + BUT; n = 8), and xylazine and butorphanol (XYL + BUT; n = 8). Two Schirmer I tear tests were also performed 15-25 min apart in dogs which received no sedative drugs (control; n = 4). Tear production rate decreased to 15 +/- 2, 17 +/- 1, and 6 +/- 1 mm min-1, respectively, while control animals averaged 21 +/- 2 mm min-1 at the same time point. Because XYL + BUT profoundly decreased tear production rate, we evaluated the two drugs separately. While BUT mildly decreased tear production when given alone to dogs (18 +/- 1 mm min-1; n = 5), xylazine had no effect on tear production. Thus it appears that the two agents act synergistically to decrease tear production rate in dogs. Moreover, sterile ocular lubricant or tear replacement should be used during XYL + BUT sedation. PMID- 11397212 TI - The effects of professional activity on the number and types of refereed publications by ACVO diplomates (1990-97). PMID- 11397213 TI - Feline ocular toxoplasmosis. AB - Ocular infection with Toxoplasma gondii is a well-recognized and important clinical entity in many animal species. In the cat, ocular toxoplasmosis is commonly associated with systemic infection, yet its role in causing anterior uveitis in an otherwise healthy cat is unclear. The purpose of this article is to review the salient epidemiological, clinical, and histopathologic features of systemic and ocular toxoplasmosis in the cat. Additionally, pathogenesis and possible immunopathogenic mechanisms of ocular toxoplasmosis, which may account for the higher prevalence of anterior uveitis in cats seropositive for T. gondii, are discussed. Finally, diagnosis, treatment and prevention of feline toxoplasmosis are reviewed. PMID- 11397214 TI - Carprofen inhibition of flare in the dog measured by laser flare photometry. AB - The purpose of this study was to determine whether oral carprofen (Rimadyl(R)) treatment in dogs could prevent or decrease the breakdown of the blood-aqueous barrier. The topical pilocarpine irritative model was used to induce breakdown and cause flare. Pilocarpine was instilled in both eyes of seven dogs at time zero and again 5 h later. At 7 h, laser flare photometry was used to measure the flare concentration in each eye using the Kowa FC-1000 laser flare cell meter. All treatments were then discontinued. Two days later, carprofen was administered to the same dogs for a total of three doses. After the last dose of carprofen, pilocarpine treatments and flare measurements were repeated. Carprofen pretreatment resulted in a 68% inhibition of flare, which was highly significant (P < 0.01). The pilocarpine group had a mean of 16.1 photon counts per millisecond (PC ms-1) +/- 2.2 SE, and the carprofen group had a mean of 7.0 PC/ms +/- 1.2 SE. These results compare favorably with previous studies measuring increased protein or fluorescein concentrations in the aqueous humor after blood aqueous barrier breakdown in the dog. These results suggest that carprofen may be effectively used as a systemically administered ocular anti-inflammatory drug. Carprofen has the added benefit of fewer reported side effects. PMID- 11397215 TI - Pectinate ligament dysplasia (PLD) and glaucoma in Flat Coated Retrievers. I. Objectives, technique and results of a PLD survey. AB - Pectinate ligament dysplasia (PLD) in dogs is a congenital ocular abnormality affecting the iridocorneal angle, the presence of which may be associated with adult-onset primary glaucoma. This paper describes the objectives, methods and results of a survey of PLD in Flat Coated Retrievers in the UK, including the technique used for gonioscopic assessment of the degree of PLD. In a random sample of 389 Flat Coated Retrievers, 34.7% were found to have a degree of PLD. In a 'target' sample of 48 Flat Coated Retrievers (close relatives of dogs from the random sample with high degrees of PLD, or dogs presented with primary glaucoma), 83.3% were found to have a degree of PLD. The percentage of animals with a degree of PLD was only 6.0% in a 'control' sample of 100 dogs from other breeds. The relationship between PLD and canine glaucoma is discussed. PMID- 11397216 TI - Pectinate ligament dysplasia and glaucoma in Flat Coated Retrievers. II. Assessment of prevalence and heritability. AB - Pectinate ligament dysplasia (PLD) in dogs is a congenital ocular abnormality affecting the iridocorneal angle, the presence of which may be associated with adult-onset primary glaucoma. This study demonstrates a significant association between PLD and glaucoma in Flat Coated Retrievers, and also a higher prevalence of PLD in Flat Coated Retrievers compared with other breeds. It is shown that the association between PLD and glaucoma was robust and not due to a small increase in PLD with age. There was also a significant association between PLD in offspring and parents and, using the offspring-parents regression, the heritability of PLD was estimated as approximately 0.7. As glaucoma is significantly related to PLD, glaucoma may also be heritable. Thus, glaucoma can be controlled by gonioscopic examination and elimination of animals with a high degree of PLD from a breeding program. Breeding from only those animals with a moderate or low PLD value or score should limit the expected prevalence of glaucoma in the resulting population to < 0.2%. The preceding study describes in detail the PLD survey, gonioscopic technique and some initial results obtained prior to this epidemiological investigation. PMID- 11397218 TI - Distribution of intraocular pressure in dogs. AB - Intraocular pressure (IOP) was measured by four different applanation tonometers in normal dogs. By MacKay-Marg tonometry in 391 dogs (772 eyes) the mean +/- SD IOP was 18.8 +/- 5.5 mmHg (range 8-52 mmHg). Using Tono-Pen XL tonometry in 421 dogs (823 eyes) the mean IOP was 19.2 +/- 5.9 mmHg, and the range was 4.42 mmHg. With MMAC-II tonometry in 80 dogs (158 eyes), the mean IOP was 15.7 +/- 2.8 mmHg with a range of 10-30 mmHg. By pneumatonograph tonometry in 135 dogs (255 eyes), the mean IOP was 22.9 +/- 6.1 mmHg and the range was 10-47 mmHg. In this study 53 breeds were represented. Of those breeds with six animals or more, no significant differences were detected in IOP between breeds (P > 0.353) or sex (P > 0.270). There was a significant decline of 2-4 mmHg (P > 0.0001) in IOP as age increased from less than 2 years to greater than 6 years of age. This trend was present with all of the four tonometers. There were no significant differences between the MacKay-Marg and TonoPen-XL tonometers (P > 0.198), but significant differences with the MMAC-II (P > 0.001) and pneumatonograph (P > 0.001) tonometers existed compared to the first two instruments. Based on this study and the literature, the mean IOP for the normal dog is 19.0 mmHg with a range of 11 (5%) and 29 (95%) mmHg. PMID- 11397217 TI - Primary lens luxation in the Chinese Shar Pei: clinical and hereditary characteristics. AB - A pedigree analysis of a family of 15 related Chinese Shar Peis was conducted. This pedigree analysis, including affected and nonaffected dams, sires and offspring, was compiled to document and characterize the occurrence, common clinical signs, and age of onset of primary lens luxation while suggesting a possible mode of inheritance in this breed. Of the five offspring from the mating of an affected dam to two unrelated affected males, 100% of offspring were affected with bilateral primary lens luxations. Of the four viable offspring from the mating of the same affected dam to an unrelated, unaffected male, two dogs (50%) were affected. The average age of onset of affected animals (seven) in this first generation was 4.9 years (range 3-6 years). The six dogs in the second generation of the same pedigree line were 2-years-old at examination with none of these animals affected at the time of this study. The most common presenting complaints were a unilateral change in ocular appearance (5 of 7 dogs) and subjective vision impairment (4 of 7 dogs). The most common clinical sign upon ophthalmic examination was iridodonesis (unilateral 4 of 7 dogs; bilateral 3 of 7 dogs) and the presence of an aphakic crescent (3 of 7 dogs). Gonioscopy and tonometry of severely affected eyes revealed a narrow or closed iridocorneal angle and ocular hypertension. This study suggests that primary lens luxation does occur in the Chinese Shar Pei, resembling the clinical condition (age of onset, clinical signs) previously described in the terrier breeds, the Border Collie, and the Tibetan Terrier. Application of the phenotypic findings in this study to a Mendelian genetic model of inheritance suggests that primary lens luxation in the Chinese Shar Pei is inherited as a simple autosomal recessive trait. PMID- 11397219 TI - Effects of metaraminol bitartrate on intraocular pressure in dogs under halothane anesthesia. AB - The effects of metaraminol bitartrate on intraocular pressure (IOP) were studied in dogs anesthetized with halothane. Forty-five healthy, adult, mixed-breed dogs, of both sexes, were divided into three groups of 15 dogs each (GI, GII and GIII) and maintained under general anesthesia with halothane after tranquilization with levomepromazine and induction with thiopental. Saline (0.9%) was administered intravenously (IV) to GI through continuous infusion, at a velocity of 0.125 mL kg-1 min-1. GII and GIII received metaraminol 0.004% IV, at a dose of 5 &mgr;g kg 1 min-1, at 0.125 mL kg-1 min-1 and at a dose of 2 &mgr;g kg-1 min-1, at 0.06 mL kg-1 min-1, respectively. IOP was measured by applanation tonometry (Tono-Pen) before and during anesthesia. Results showed that IOP decreased in GI, increased in GII, and remained at basal levels in GIII. Continuous infusion of metaraminol at 2 &mgr;g kg min-1 maintained IOP at pretest levels, while infusion at 5 &mgr;g kg-1 min-1 produced an elevation of IOP. PMID- 11397220 TI - The use of xenologous amniotic membrane to repair canine corneal perforation created by penetrating keratectomy. AB - This study was performed to evaluate the use of glycerol-preserved equine amniotic membrane as replacement for full-thickness corneal defects in dogs. Eighteen mixed-breed dogs were used. A perilimbal, full-thickness, 5 mm square corneal defect was created surgically, and a donor implant of equine amniotic membrane of the same size and shape sutured in place with 10-0 nylon simple interrupted sutures. Corneal edema was observed near the implant 24 h after surgery, but was absent after 1 week. Granulation tissue and corneal vascularization superficial to the implant were noticed on postoperative day 7, but were absent on day 30. Corneal vascularization persisted until the end of the experiment. There was no fluorescein retention by postoperative day 30. There was slight clearing of the corneal implant by postoperative 30, and slight pigmentation of the donor implant observed at postoperative day 180. An acute inflammatory process as well as fibroblasts were present at early postoperative stages. At postoperative day 60 there was no inflammatory cellular infiltrate, but fibroblasts and fibrosis were present. Corneal architecture was restored at the end of the experiment, with a layering of the epithelium-stroma-debris of amniotic membrane-stroma-endothelium present, and pigmentation and vascularization present in the deep layers of the cornea. Although vascularization indicated some degree of graft rejection, the clinical and histological evidence indicates that the xenologous amniotic membrane can be useful as a tectonic graft in the repair of full-thickness lesions of the cornea of dogs. PMID- 11397221 TI - Respiratory function and extraocular muscle paralysis following administration of pancuronium bromide in dogs. AB - Pancuronium bromide, a neuromuscular blocking agent, was evaluated in canine cataract surgical patients under general anesthesia to determine its effects on respiratory function and globe position. Two paralytic, anesthetic regimes were studied: one using a standard dosage of 0.066 mg kg-1 pancuronium bromide, given intravenously while providing the patient with ventilatory support, and one using a dosage of 0.022 mg kg-1 in which no ventilatory support was provided. Eye position and anterior vitreal position/displacement were recorded by a surgeon who was blinded as to treatment group. Physiological parameters indicative of respiratory function were monitored. Both dosages of pancuronium produced comparable, neutral globe position within 30 s following administration which lasted for 20-30 min. All patients in the standard dose group experienced uneventful anesthetic episodes with physiological parameters well within the normal ranges. Within 5 min after administration, all patients in the low-dose group developed a pronounced respiratory acidosis (mean arterial pH = 7.07 +/- 0.08; mean PaCO2 = 79.8 +/- 10.7 mmHg), which exceeded a set of predetermined safety limits, and subsequently these dogs received ventilatory support. We conclude that 0.022 mg kg-1 pancuronium rapidly produces an unacceptable level of respiratory acidosis and, as a result, patients receiving neuromuscular blocking agents should routinely receive ventilatory support. PMID- 11397222 TI - Histomorphometry of the porcine scleral lamina cribrosa surface. AB - The lamina cribrosa surface of the pig was examined using trypsin digestion, scanning electron microscopy, and computerized image analysis. Six normal eyes from healthy pigs were studied. The total intralaminar scan area, total number of laminar pores, median individual laminar pore areas, median individual pore form factors, and mean pore density were determined for the dorsal, ventral, nasal and temporal hemicircles, the dorsal-nasal, dorsal-temporal, ventral-nasal, and ventral-temporal quadrants, and their central and peripheral subdivisions. The mean (+/- SD) total intralaminar scan area was 8.29 +/- 1.54 mm2. The mean (+/- SD) total laminar pore count was 517 +/- 73 pores. The mean pore count was significantly larger in the ventral than the dorsal hemicircle (292 +/- 39 vs. 225 +/- 38 pores, respectively; P = 0.001), and significantly greater in the periphery compared to the center (388 +/- 58 vs. 129 +/- 27 pores, respectively; P = 0.0001). The overall mean (+/- SD) pore density was 67 +/- 7 pores mm-2. Mean pore density was significantly greater in the ventral than the dorsal hemicircle (70 +/- 8 vs. 64 +/- 6 pores mm-2, respectively; P = 0.019), and significantly greater in the center compared to the periphery (75 +/- 9 vs. 60 +/- 8 pores mm 2, respectively; P = 0.020). The mean (+/- SD) median individual pore area was 3752 +/- 572 &mgr;m2. The mean (+/- SD) median pore form factor was 0.680 +/- 0.035. No significant regional differences were found in mean median pore form factor or mean median individual pore areas. The intralaminar optic nerve of pigs is 55.1% non-neural connective and vascular tissue. A pigmented ventral fascial groove in the scleral lamina cribrosa appears unique to the porcine lamina cribrosa, and may be a vestige of the embryonic optic fissure. PMID- 11397223 TI - Surgical management of equine recurrent uveitis with single port pars plana vitrectomy. AB - Current information suggests that equine recurrent uveitis (ERU) is an immune mediated reaction to infectious agents or to autologous ophthalmic tissue. Recurrences are associated with progression of irreversible ocular damage. This report describes the intraoperative technique, complications, and long-term results of 38 eyes in 35 horses with ERU that underwent pars plana vitrectomy. The majority of the horses were warm-blooded. Recurrence of ERU was prevented in 35 of the 38 eyes. Some horses, especially in patients with incipient cataracts, developed vision loss in postoperative, quiescent eyes which was usually associated with cataract formation. Vision was stable in 85% of all eyes that underwent vitrectomy. Pars plana vitrectomy in horses appears successful in interrupting the cycle of repeated episodes of ERU, and the subsequent globe destruction in the majority of eyes. Removal of uveitis-induced 'immunologic memory' in the vitreous by vitrectomy may reduce adverse interaction between the vitreous and the uveal tract, and therefore reduce the recurrence of ERU. PMID- 11397224 TI - A histopathological study of iridociliary cysts and glaucoma in Golden Retrievers. AB - This is a retrospective histopathological study of archive slides from the Comparative Ocular Pathology Laboratory of Wisconsin from 1981 to 1997. Reports on eyes or ocular contents from 2176 dogs were reviewed. Five-hundred and thirty out of 2176 (24%) of the cases had a clinical or histologic diagnosis of glaucoma. Twenty-five out of 530 (5%) of the canine cases of glaucoma were in Golden Retrievers. Thirteen out of 25 (52%) of the Golden Retriever cases of glaucoma had iridociliary cysts. Iridociliary cysts in Golden Retrievers may lead to the development of glaucoma. Histologically all 13 of the cases of glaucoma and iridociliary cysts had large thin-walled cysts lined with attenuated cuboidal epithelium filling most of the posterior chamber. The following histologic features were also present: thick walled cysts containing hyaluronic acid (8/13; 62%), a solid cellular proliferation (2/13; 15%), a retrocorneal membrane associated with a defect in Descemet's membrane (7/13; 54%), iris bombe (5/13; 38%), a preiridal fibrovascular membrane (4/13; 31%), hemorrhage (4/13; 31%), a cellular membrane on the anterior lens surface (6/13; 46%), retinal detachment (5/13; 38%), peripheral anterior synechia (5/13; 38%), and posterior synechia (6/13; 46%). Complete follow up was obtained on 11/13 (85%) of the Golden Retrievers with glaucoma and iridociliary cysts. Two/11 (18%) of the dogs were euthanized due to intractable glaucoma. Eight/13 (62%) of the dogs had problems in the other eye. The other eye was diagnosed with uveitis 5/11 (45%), glaucoma 2/11(18%), pigment in the anterior chamber 3/11 (27%), and iridociliary cysts 4/11 (36%). PMID- 11397225 TI - Intraocular pressure in normal dairy cattle. AB - Intraocular pressure (IOP) was measured in normal dairy cows by applanation tonometry. In the first study of 15 Holstein and 17 Jersey cows the mean IOP by Mackay-Marg tonometry was 27.5 +/- 4.8 mmHg (range 16-39 mmHg); no significant differences (P < 0.92) were observed between the Holstein and Jersey breeds. In the second study of 15 Holstein and 12 Jersey cows, the mean IOPs by Mackay-Marg and TonoPen-XL tonometry were 28.2 +/- 4.6 mmHg (range 19-39 mmHg) and 26.9 +/- 6.7 mmHg (range 16-42 mmHg), respectively. Comparisons of the Mackay-Marg and TonoPen tonometers indicated no significant differences (P < 0.16). The mean and range of IOP in normal dairy cows within 2 SD (95% of the population) is 27 mmHg with a range of 16-36 mmHg. PMID- 11397226 TI - Lamellar keratoplasty for the treatment of feline corneal sequestrum. AB - A lamellar keratoplasty was used to treat corneal sequestrum in four Persian cats (six eyes). Following a superficial keratectomy, lamellar corneal allografts (feline corneal tissue) or heterografts (canine corneal tissue) which had been preserved at -20 degrees C were placed in the recipient cornea. All grafts became optically transparent within 2 months following surgery and no recurrences of the sequestrum have been noted during the follow-up period (4-30 months). We conclude that feline corneal sequestrum may be successfully treated with feline or canine donor corneal tissue using this technique. PMID- 11397227 TI - Ocular involvement of transmissible venereal tumor in a dog. AB - A case of metastatic transmissible venereal tumor (TVT), with lesions on the penis, conjunctiva, buccal mucosa and skin (lips and trunk), is presented in this case report. The clinical picture is described along with the cytological and histopathological features of the tumor leading to definitive diagnosis of TVT. Possible reasons for the unusual metastatic behavior of TVT and full recovery of the dog after chemotherapy are discussed. PMID- 11397228 TI - Presumed lipid retinopathy in a diabetic dog. AB - An eight-year-old neutered male diabetic Cardigan Welsh Corgi was presented for bilateral mature cataracts. Phacoemulsification and intraocular lens implantation were performed routinely, and recovery was uneventful for several months except for lipemic aqueous flare which gradually resolved during the postoperative period. Five months following surgery the owner presented the dog for decreased vision. White retinal deposits were visualized ophthalmoscopically. Serum analysis revealed that the dog was concurrently markedly hyperlipemic. The patient's diet was changed from a high- to a low-fat diet, following which diabetes control improved, hyperlipemia resolved, and the retinal deposits decreased markedly in size. The retinal deposits seen ophthalmoscopically are presumed to be lipid based upon their association with marked hyperlipemia, and the concurrent resolution of hyperlipemia and the fundic lesions. PMID- 11397229 TI - Cyclosporine A in veterinary ophthalmology. PMID- 11397230 TI - The use of immunohistochemistry and the polymerase chain reaction for detection of feline leukemia virus and feline sarcoma virus in six cases of feline ocular sarcoma. AB - Ocular sarcoma was diagnosed by light microscopic examination in enucleated globes (n = 4), orbital tissue biopsy (n = 1) and ocular evisceration contents (n = 1) from six cats. To determine if feline leukemia virus (FeLV) or a replication defective FeLV, feline sarcoma virus (FeSV), was present in these ocular sarcomas, immunohistochemistry (IHC) and polymerase chain reaction (PCR) for FeLV were utilized. Immunohistochemical staining for FeLV glycoprotein 70 (gp70) was performed on all six formalin-fixed, paraffin-embedded tumors using an avidin biotin complex technique. DNA was extracted from each specimen and a 166 bp region of the FeLV long-terminal repeat (LTR) was amplified by PCR. All tumors were composed primarily of spindle cells; two neoplasms had PAS-positive basement membrane enveloping areas of spindle cells. All tumors involved the uvea and five of six tumors showed transcleral extension, one of which invaded the optic nerve. Immunohistochemical staining for FeLV gp 70 was negative. PCR to amplify a portion of the FeLV LTR was negative. Based on these findings of these limited number of cases, FeLV/FeSV may not play a role in the tumorigenesis of feline ocular sarcomas. However, additional tumors representing all morphological subtypes should be investigated for the presence of viral antigen and DNA. It is important to determine the etiology and pathogenesis of these malignant ocular sarcomas. If the cell of origin and pathogenesis involve ocular and lenticular injury, and FeLV/FeSV is not present, then the clinical management of cases of feline ocular trauma, uveitis and glaucoma may prevent the development of this tumor. PMID- 11397231 TI - Heterochromia iridis in water buffaloes (Bubalus bubalis). AB - This study included 45 unaffected animals and 593 animals affected with heterochromia irides, and 85 enucleated eyeballs with heterochromia irides. The classification of heterochromia irides, morphology of normal and heterochromic irides, and the histology, ultrastructure, and scanning electron microscopy are presented. The incidence of heterochromia irides in water buffaloes was 7.62% affecting either one or both eyes. Both complete and partial heterochromia irides occurred. Complete heterochromia iridis is more frequent than the partial form in either bilateral or unilateral cases. The pupil has a dumb-bell-shape appearance. Granula iridica occurred at the upper (100%) and lower (30%) pupillary margins and originated from the posterior pigmented epithelium. In heterochromia irides, the melanocytes is absent in the anterior border and stromal layers, and iridal thickness appeared thinner than that of normal eyes. PMID- 11397232 TI - Screening and controlling canine inherited ocular diseases in Finland: epidemiological, economical and health promotional aspect. AB - Official eye examination records of 18 146 dogs in 17 breeds were analyzed in order to evaluate the Finnish Kennel Club's eye examination scheme. The most common inherited ocular diseases were the Collie eye anomaly (CEA) in the Rough Collie and the persistent hyperplastic tunica vasculosa lentis (PHTVL) in the Doberman. In both diseases the prevalence has also increased during the study period 1988-1997. The most likely reason for the increase is the change in examination routines that now makes it easier to find milder disease forms that were previously classified as normal. The prevalences of other inherited diseases are so low that no conclusions about the trends can be made. Also, the data have some defects that can cause biased estimations of prevalences. The costs of screening programs are very high and no direct economical benefits can be shown. Methods of improving the screening and effectiveness of the program are also presented. PMID- 11397233 TI - Multifocal retinopathy of Great Pyrenees dogs. AB - Forty-four related Great Pyrenees dogs were examined ophthalmoscopically. Focal retinal elevations, multiple gray-tan-pink subretinal patches, and discrete areas of tapetal hyper-reflectivity were seen in 19 dogs, ranging from 13 weeks to 10 years of age. These lesions varied in size from focal spots that were barely visible with the indirect ophthalmoscope to areas that were larger than the optic disc. Complete blood cell counts, serum biochemical profiles, urinalyses, and blood pressure measurements were completed on four affected dogs and all were within normal reference ranges. Photopic and scotopic electroretinography was completed and the a-wave and b-wave amplitudes and latencies were similar for affected and age-matched nonaffected Great Pyrenees and other normal dogs. Electroretinograms that were examined twice during a 3-year period on three affected adult dogs did not reveal significant progressive deterioration of the a or b-wave parameters. Fluorescein angiography was completed on four affected dogs of ages 1 (n = 2), 5, and 6 years. These angiograms were repeated in three of these dogs 1 year later. The blood ocular barrier was intact in these dogs but there was blocked choroidal fluorescence. Postmortem examination, light microscopy, scanning and transmission electron microscopy were performed on three affected puppies and two affected adult dogs. These examinations revealed that the lesions in the puppies were limited to bilateral multiple areas of retinal pigment epithelial vacuolation, hypertrophy, and apparent separation from Bruch's membrane, and multiple serous retinal detachments. The affected adult dogs had focal retinal degeneration and retinal pigment epithelial hypertrophy, hyperplasia and pigmentation. Pedigree analysis and test mating confirm that this condition is inherited, probably as an autosomal recessive trait. This condition develops at approximately 13 weeks of age and the focal areas of retinal detachment and retinal pigment epithelial vacuolation progress to permanent and stable focal areas of retinal degeneration, and retinal pigment epithelial hypertrophy and pigmentation. PMID- 11397234 TI - Iridociliary epithelial tumors in 100 dogs and 17 cats: a morphological study. AB - The morphological features of iridociliary epithelial tumors in 100 dogs and 17 cats were reviewed. Twenty-seven cases were in either Golden Retrievers or Labrador Retrievers. Affected globes were stained for light microscopy with alcian blue, periodic acid Schiff (PAS) and hematoxylin and eosin stains. Selected tissues were examined by immunohistochemistry for vimentin, desmin, cytokeratin, S-100, neuron-specific enolase (NSE), and glial fibrillary acid protein (GFAP). The presence or absence of hyaluronic acid was recorded by staining with alcian blue before and after digestion of the tissue with hyaluronidase. Canine tumors were divided into papillary and solid tumors based on the pattern of growth. Twenty-eight of 57 papillary tumors exhibited invasive behavior including eight of the 57 which showed infiltration of the sclera. Twenty-nine of 43 solid tumors were invasive including 13 of 43 with scleral invasion. Tumors with scleral invasion were designated adenocarcinoma. Tumors of both types could be pigmented or nonpigmented and often contained smooth basement membranes reminiscent of the inner membrane of the nonpigmented ciliary body epithelial cell. All of the feline tumors were nonpigmented and 14 of 16 feline tumors were solid and two of the tumors were papillary. Eighteen of 20 canine tumors and three of four feline tumors stained positive for vimentin. Cytokeratin stain was positive only in a few of the highly aggressive tumors. The finding of pigmented epithelial cells, thick, smooth basement membrane structures, positive staining for vimentin, S-100, and NSE as well as hyaluronic acid deposition were considered to be features which define iridociliary epithelial tumors in dogs. The positive staining for vimentin and NSE are highly specific markers which help to characterize iridociliary epithelium and distinguish this tumor from metastatic epithelial tumors. The finding of solid nonpigmented tumors with small epithelial cells packeted by thin PAS-positive membranes staining positive for vimentin were considered significant features defining iridociliary epithelial tumors in cats. Follow-up information on survival and cause of death was obtained on 43 canine cases and only two feline cases. The average follow-up interval in dogs was 25 months and only two dogs died with lesions that could have been due to metastasis although neither was confirmed. We concluded that neither iridociliary adenomas nor adenocarcinomas is likely to metastasize. PMID- 11397235 TI - Results of grid keratotomy, superficial keratectomy and debridement for the management of persistent corneal erosions in 92 dogs. AB - Ninety-two cases of persistent corneal erosions in dogs were analyzed for breed, gender, age and which eye was affected. The results of the treatment of 92 persistent corneal erosions in dogs by superficial keratectomy (SK), grid keratotomy (GK), or debridement with a sterile dry cotton swab are presented. These techniques gave better rates of healing than have been previously reported. All cases of persistent corneal erosions healed in this study. However, it must be noted that three cases treated with debridement only failed to heal after several treatments and were eventually treated with SK. After one procedure 80 out of 92 (87%) had healed. After one procedure, 63% of cases treated with debridement healed, 100% of cases treated with SK healed, and 85% of cases treated with GK healed. At the first postoperative visit, 88% (21/24 cases) of ulcers treated by SK had healed, and 75% (39/52 cases) of ulcers treated by GK had healed. Only 25% of the persistent corneal erosions had healed at the first visit after debridement. All 24 cases of persistent corneal erosions treated with SK healed after one treatment in a mean +/- SD of 9.3 +/- 3.9 days (median of 7 days). Fifty-two cases were managed with GK; 44 (83%) of these healed with one procedure and eight cases required a second GK procedure to resolve. A mean +/- SD of 13.4 +/- 5.1 days (median of 11.5 days) following GK was required for the persistent corneal erosions to heal. Nineteen cases were initially managed by debridement with a dry cotton swab under local anesthesia. Sixteen out of these 19 debridement cases healed (giving an overall healing rate of 84%) in a mean +/- SD time of 23.4 +/- 11.1 days (median 21.5). There were three cases that did not heal with debridement. These cases were debrided at 10-20 day intervals for 30-60 days, and were then treated with SK. Two of these cases healed within 7 days, the other case required 18 days to heal. Sixty-three per cent of persistent corneal erosions treated with debridement healed after one procedure; however, only four out of 19 cases (21%) were healed at the first revisit. Complications were rare: corneal edema occurred in two cases following multiple GK, and excessive granulation tissue in one case was managed with a SK. There was the occurrence of an ulcer adjacent to the surgery site in four cases, two cases following GK and two cases following SK. PMID- 11397236 TI - Clinical, cytologic and histopathologic manifestations of protothecosis in two dogs. AB - Two cases of sudden onset of blindness associated with ocular protothecosis in dogs are reported. Both were adult, spayed female, mixed-breed dogs that lacked the usual clinical signs of systemic infection with Prototheca species. Physical abnormalities at the time of presentation were limited to the affected eyes which had serous discharge, hyperemic conjunctiva, and aqueous flare. The pupillary light reflexes were slow, and the menace reflexes were absent. Both dogs had glaucoma. Results of complete blood counts and serologic titres for antibodies to Blastomyces dermatitidis and Histoplasma capsulatum were within reference intervals. Protothecosis was diagnosed by cytologic analysis of vitreous humor and was confirmed at necropsy. These two cases were unusual because of their presenting signs and prolonged course of disease progression. PMID- 11397237 TI - Where Now? PMID- 11397238 TI - Comparative retinal ganglion cell and optic nerve morphology. AB - The optic nerve is divided in four regions: intraocular, intraorbital, intracanalicular, and intracranial. The vertebrate retinal ganglion cells are classified by morphology, physiology and soma size. Species differences and similarities occur with retinal ganglion cells. Alpha retinal ganglion cells have large somata, large dendritic fields, large-diameter axons, and are most dense in the peripheral retina. Beta retinal ganglion cells have smaller diameter somata, smaller dendritic fields, small diameter axons, and predominate in the central retina. Gamma retinal ganglion cells are a heterogenous class of cells and have small diameter axons, and slow axon conduction velocities. The spatial distribution and organization of the retinal ganglion cells extends retinotopically through the nerve fiber layer, optic nerve, optic chiasm, optic tract, lateral geniculate nucleus, and visual cortex. The retinal nerve fiber layer thickness decreases from the optic disk toward the periphery of the retina. The retrobulbar optic nerve axon counts and axon density vary by species, with larger nerves having higher axon counts. Decussation of the optic nerve axons at the optic chiasm varies with 100% decussation in most birds and fish, 65% in cats, 75% in dogs, 80-90% in large animals, and 50% in primates. Centrifugal axons also occur in the optic nerve and may represent a method by which the brain can influence retinal activity. PMID- 11397239 TI - Comparative optic nerve physiology: implications for glaucoma, neuroprotection, and neuroregeneration. AB - The axoplasm of optic nerve axons moves bidirectionally at various speeds along an intra-axonal pressure gradient from the retinal ganglion cell (RGC) somata toward its synapse, and from the synapse towards the RGC somata. The axoplasmic flow of optic nerve axons is precarious even at normal intraocular pressures (IOP) as it moves from the intraocular optic nerve through the scleral lamina cribrosa to the intraorbital optic nerve. The scleral lamina cribrosa is not simply a porous region of the sclera but a specialized extracellular matrix of the central nervous system whose movement during fluctuations in IOP can affect optic nerve axoplasmic flow. The abundant optic nerve blood supply maintains adequate optic nerve head perfusion through a process of vascular autoregulation. Glaucoma is associated with reduced optic nerve axoplasmic flow and compromised optic nerve circulation such that RGC death due to glutamate excitotoxicity and neurotrophin deprivation result. PMID- 11397240 TI - The influence of aging and low zinc nutrition on the choroid in the pig: I. The melanocyte. AB - The effects of low zinc nutrition and aging on central choroidal melanocytes were examined in the pig. Three populations of pigs (young, pregnant and aged), were maintained on either control (C) or low zinc (LZ) diets. Twenty-five weanling boars were sacrificed at 2, 4, 8, 10, and 12-month intervals, and nine pregnant sows and eight aged sows were sacrificed after a 6-month interval. Melanocytes of the central choroid were morphologically and morphometrically examined. The melanocyte was found to be conservative in its form, which was mostly elliptical longitudinal profile, throughout the different age populations that were fed the C diet. Morphometric observations revealed that this cell type increased in size in the oldest animals, having been 40% greater than that in the younger two populations. However, the overall percentage area occupied by melanocytes remained the same throughout all age groups. In the animals that were fed the LZ diets, a large subpopulation of choroidal melanocytes was oval to round in shape in the pregnant and aged groups. Many members of this subpopulation possessed less opaque pigment than the elliptically shaped cell. Measurements of the size and percentage area occupied in these oldest groups increased significantly. In addition to the change of size, shape and melanin opaqueness, unusually large melanosomes were consistently observed in the pregnant and aged LZ groups. Low zinc nutrition had a remarkable age-related impact on the usually quiescent melanocyte. PMID- 11397241 TI - The influence of aging and low zinc nutrition on the choroid in the pig: II. The melanosome. AB - The effects of low zinc nutrition and aging on central choroidal melanosomes were examined in the pig. Melanosomes of central choroidal melanocytes were morphologically and morphometrically examined in three populations of pigs (young, pregnant and aged), that were maintained on either control (C) or low zinc (LZ) diets. In C groups, the typical melanosomes decreased in size with age, although a subpopulation of larger melanosomes occurred in the oldest group. In contrast, the melanosomes of the animals on LZ diets increased in size significantly in the adult groups. The melanosomes in the pregnant and aged groups were 65% and 30-40% greater than those of the age-matched C groups. Extremely large melanosomes were frequently encountered in adult LZ choroidal melanocytes. Melanogenesis of these large bodies included the formation of one or more outer shells. Fusion of adjacent large melanosomes was also observed. Melanolysosomal-like bodies were observed, particularly among the pigmented cells in the large blood vessel region of C and LZ adults. Melanin dynamics, i.e. its production and breakdown, occurred within choroidal melanocytes throughout much of the lifespan of the pig. This dynamic was greatly influenced by low zinc nutrition, resulting in unusual and aberrant melanin activity. PMID- 11397242 TI - Congenital ocular abnormalities of Rocky Mountain Horses. AB - OBJECTIVE: To determine the incidence and describe ocular abnormalities in a cross-section of the population of Rocky Mountain Horses. Design: Prospective study. Animals: Five-hundred and fourteen Rocky Mountain Horses. Procedure: Ophthalmic examinations were performed using a slit-lamp biomicroscope and an indirect ophthalmoscope. Intraocular pressures were measured by applanation tonometry. Eyes from six horses were obtained for histologic examination. RESULTS: Cysts of the posterior iris, ciliary body, and peripheral retina were detected most frequently (249 horses), and were always located temporally. Curvilinear streaks of retinal pigmented epithelium extending from the peripheral temporal retina marked the boundary of previous retinal detachment in 189 horses. Retinal dysplasia was detected in 125 horses. Multiple ocular anomalies were evident in 71 horses and were always bilateral and symmetrical. Affected eyes had a large, clear cornea that protruded excessively and had an apparent short radius of curvature, a deep anterior chamber, miotic and dyscoric pupil, and iris hypoplasia. Pupillary light responses were decreased or absent and pupils failed to dilate after repeated instillation of mydriatic drugs in horses with multiple ocular anomalies. Less frequently encountered abnormalities included peripheral iridocorneal adhesions and goniosynechiae. Congenital cataract was always present in eyes with multiple abnormalities. Intraocular pressures did not differ among horses with normal eyes and horses with multiple ocular abnormalities. Histologic examination of eyes corroborated the clinical appearance. PMID- 11397243 TI - The geographic form of retinal dysplasia in dogs is not always a congenital abnormality. AB - To examine the congenital nature of the geographic form of focal/multifocal retinal dysplasia, we carried out a retrospective analysis of the medical records of dogs produced in a closed colony of service dogs who receive very thorough ophthalmologic examinations early in their life, and later, when they return for training. Medical records were reviewed from all dogs produced by The Seeing Eye, Inc. between October 1991 and September 1998, and which had a diagnosis of geographic retinal dysplasia coded. We identified 23 dogs of five different breeds or interbreed crosses that comprise the breeding and production program (Golden Retrievers, German Shepherds, Labrador Retrievers, Labrador Retriever/Golden Retriever cross and German Shepherd/Labrador Retriever cross) in which the results of at least two complete ophthalmic examinations were documented, the first before 10 weeks of age, and the second when the dog was a young adult. Of the 23 dogs, only one was identified as affected with the geographic form of retinal dysplasia when examined at 5-6 weeks of age. The remaining dogs were normal. Our findings indicate that, in most cases, the geographic form of retinal dysplasia is not present in dogs prior to 10 weeks of age. These findings indicate the need to revise recommendations for early screening of dogs for retinal dysplasia. PMID- 11397245 TI - Retinal dystrophy or 'congenital stationary night blindness' in the Briard dog. PMID- 11397244 TI - Clinical, electrophysiological and morphological changes in a case of hereditary retinal degeneration in the Papillon dog. AB - An autosomal recessive retinal disease with a late onset in Swedish Papillon dogs has recently been described. A 7-year-old Papillon dog showed no obvious signs of visual impairment and only minor ophthalmoscopic changes. Cone ERG b-wave amplitudes were within normal limits, while rod responses were nonrecordable or severely abnormal. Ultrastructural examination showed a generalized retinal degenerative disease, most prominent in the peripheral areas. The inferior retina was more severely affected than the superior areas. Both rods and cones showed morphological changes. The Papillon dog is another dog breed affected by progressive rod-cone degeneration, with similarities to the canine retinal disease given the gene symbol prcd. PMID- 11397246 TI - The use of preserved equine renal capsule to repair lamellar corneal lesions in normal dogs. AB - The purpose of this study was to evaluate the use of equine renal capsule preserved in 98% glycerine to repair lamellar corneal lesions in normal dogs. For this purpose, 12 dogs, divided into six groups (n = 2), were used to evaluate the 1st to 7th day, 15th day and 30th to 60th postoperative day. In order to perform the histologic study, the clinical procedures were analyzed, while the recipient's corneas were collected. The photophobia and blepharospasm also were more intense in the 1st to 7th postoperative day, and regressed in the 15th postoperative day. Therefore, the edema and the vascular events were both more frequent in the intermediary phases and regressed in the late periods. On the other hand, the morphological evaluation demonstrated an inflamatory exudate, also in the intermediary and late periods. These results suggested that the equine renal preserved capsule could be a useful alternative tissue to repair lamellar corneal lesions in dogs. PMID- 11397247 TI - Antioxidant profile of cataractous English Cocker Spaniels. AB - Cataracts have been attributed to oxidative injury in proteins and lipids. Primary defenses that directly protect the lens against oxidative damage include small molecule antioxidants (vitamin C, vitamin E, glutathione and carotenoids) and antioxidant enzymes (superoxide dismutase, catalase, and the glutathione enzyme systems - glutathione peroxidase, glutathione reductase and glucose-6 phosphate dehydrogenase). In humans, low plasmatic levels of vitamin C, vitamin E and carotenoids have been associated with a high risk of senile cataracts. Dogs are more prone to develop cataracts. A decrease in antioxidant defenses could be responsible for increased lens oxidation and cataract development. In this study we report the levels of erythrocytic enzymatic antioxidants (superoxide dismutase, catalase, glutathione peroxidase and glucose-6-phosphate dehydrogenase) and plasma vitamin C as well as malondialdehyde, in normal and cataractous English Cocker Spaniel dogs. Plasma vitamin C levels were consistently lower in cataractous dogs (20.17 &mgr;M +/- 8.2 &mgr;M) when compared with normal dogs (24.1 &mgr;M +/- 9.4 &mgr;M). These results indicate a possibly decreased synthesis in vitamin C, leading to lower aqueous humor levels of this vitamin. Considering that vitamin C levels in the aqueous humor may be responsible for lens antioxidant maintenance, and that these levels are obtained from plasma secretion through the ciliary epithelium, decreased plasma levels may indicate a decrease in the antioxidant capacity of the aqueous humor. PMID- 11397248 TI - Doppler imaging of the ophthalmic vasculature of the normal dog: blood velocity measurements and reproducibility. AB - The purpose of this study was to assess the feasibility and reproducibility of color Doppler imaging (CDI) of the vasculature of the normal canine orbit and eye. Eight normal Beagles were evaluated by Doppler imaging. The goals of the study were to determine the location, spectral waveform morphology, specific blood velocity parameters, and reproducibility for the ophthalmic and orbital vessels most frequently identified in the normal dog. Vessels identified a majority of the time (> 50%) included: external ophthalmic artery, dorsal external ophthalmic vein, ventral external ophthalmic vein, internal ophthalmic artery, anterior ciliary artery and vein, short and long posterior ciliary arteries, primary retinal arteries, and vortex veins. Other vessels imaged less frequently included: external ethmoidal artery (50%), and primary retinal veins (25%). For each blood vessel the time averaged velocity, peak systolic velocity, minimum diastolic velocity, pulsatility index, and resistive index were determined. The ophthalmic and orbital vessels have unique spectral waveforms and velocities which serve as a basis for identification. Reproducibility of the most commonly imaged vessels of the canine eye and orbit with Doppler imaging was high (< 10% variation). Doppler imaging has the potential for determining noninvasively and consecutively the blood velocity parameters found in orbital and ocular diseases, including orbital inflammations and neoplasms; intraocular inflammations and neoplasms; vascular diseases including systemic vascular disease (hypertension), vasculopathies, and anemia; the glaucomas; and documentable follow-up after medical and/or surgical treatment of these diseases. PMID- 11397249 TI - Comparative Doppler imaging of the ophthalmic vasculature in normal Beagles and Beagles with inherited primary open-angle glaucoma. AB - The objective of this study was to compare orbital and ocular vasculature velocity, measured by Doppler imaging, in normal Beagles and Beagles with inherited primary open-angle glaucoma. Eight normal Beagles and 13 Beagles with different stages of primary open-angle glaucoma were evaluated twice with a 2-4 week period between measurements. Doppler imaging was performed with the dogs anesthetized, and the Doppler transducer applied directly on the corneal surface. The majority of the orbital vasculature (external ethmoidal artery; internal ophthalmic artery and vein; and external ophthalmic artery and vein) and ocular blood vessels (anterior ciliary artery and veins; long posterior ciliary arteries; short posterior ciliary arteries; primary retinal arteries; and the vortex veins) were identified and Doppler blood velocity parameters were determined. The glaucomatous dogs demonstrated significant differences in the Doppler velocity parameters of several orbital vessels (external ethmoidal, external ophthalmic, and internal ophthalmic arteries), and several ocular vessels (anterior ciliary, short posterior ciliary, and long posterior ciliary arteries). These differences included decreased blood velocities, and increased pulsatility and resistive indexes. The Doppler blood flow velocities of the primary retinal arteries were unchanged between the normal and glaucomatous dogs. In the glaucomatous dogs, the Doppler imaging suggests increased vascular resistance downstream in both the orbital and ocular vasculature. These blood velocity parameter changes may be primary or secondary, and may offer therapeutic opportunities to increase perfusion, prolong the retina and optic nerve head function, and maintain vision in the canine glaucomas. PMID- 11397250 TI - Vasculature of the orbital rete in the Japanese deer (Cervus nippon). AB - The vasculature of the orbital rete (rete mirabile ophthalmicum) in Japanese deer (Cervus nippon) was studied using corrosion casting, scanning electron microscopy, and histology. The orbital rete is a flat, triangular- or leaf-shaped arterial network, which consists of a complex of small arterioles, that intermixes with a similar complex of the supraorbital vein at the base of the orbital cavity. Blood to the retina passes through the orbital rete. The orbital retial arterioles leave the parent external ophthalmic artery at right angles forming T-shaped bifurcations, and follow a tortuous, undulating course. Each retial arteriole is connected by side branches and forms a rope-ladder-like network. Some of the side branches are surrounded by a groove representing the intra-arterial cushion that regulates blood flow at branching sites. The central retinal artery supplying the retina originates from the orbital rete. The ciliary arteries supplying the choroid arise from the external ophthalmic artery proximal to the orbital rete. The anatomical specializations of the orbital rete may involve buffering the blood pressure and flow to the retina and regulating ocular tissue temperature as in the carotid rete. In addition, the orbital rete may help dampen the tension that the vessel exerts on the retina, by stretching in response to eyeball movement. PMID- 11397251 TI - Use of nested polymerase chain reaction (PCR) for detection of retroviruses from formalin-fixed, paraffin-embedded uveal melanomas in cats. AB - Thirty-six formalin-fixed, paraffin-embedded enucleated globes from cats with a diagnosis of diffuse anterior uveal melanoma were obtained. Sections of tumor were excised, deparaffinized, and subjected to nested polymerase chain reaction (PCR) to identify proviral DNA sequences from the feline leukemia virus (FeLV) feline sarcoma virus (FeSV; 36 eyes), and the feline immunodeficiency virus (FIV; 18 eyes). All samples tested were negative for FIV DNA. Three samples were positive for FeLV-FeSV DNA. This is the first reported evidence of a possible link between naturally occurring feline anterior uveal melanoma and the presence of FeLV-FeSV DNA. PMID- 11397252 TI - Lymphosarcoma with conjunctival manifestation in a guinea pig. AB - A guinea pig with multicentric lymphosarcoma and conjunctival manifestation is described. Primary clinical signs were bilateral infiltrative conjunctival masses. Antemortem diagnosis was based on the cytology of biopsies of conjunctival tissue, fine-needle aspirates of peripheral lymph nodes, and peripheral blood smears. PMID- 11397253 TI - Atypical primary ocular melanoma in cats. AB - The clinical and pathologic features of six cases of atypical primary ocular melanoma in cats are described. In contrast to diffuse iris melanomas, these atypical tumors originated multifocally from any portion of the uvea and in five of the six cases were very advanced at the time of clinical presentation. Two of the advanced cases involved primarily the ciliary body, while three others showed extensive involvement of the choroid. Histologically, these tumors consisted primarily of round, heavily pigmented cells lacking the anaplastic nuclear features characteristic of advanced diffuse iris melanomas. The advanced stage of the tumors at presentation may have been due to either the posterior uveal origin of these tumors or to more rapid progression. Additional cases with long-term follow-up need to be studied in order to accurately assess the biological behaviour of these tumors. PMID- 11397254 TI - Angioinvasive pulmonary carcinoma with posterior segment metastasis in four cats. AB - The objective of the research was to characterize the clinical, fluorescein angiographic, pathologic and microscopic features of feline pulmonary carcinoma with ocular metastasis that resulted in ischemic chorioretinopathy. Four cats with confirmed or presumed primary pulmonary neoplasia with posterior segment metastasis were studied. The medical records from four cats with a diagnosis of bronchogenic carcinoma and intraocular metastasis were reviewed. Physical and ophthalmic examinations and thoracic radiographs were performed in all cases, and fluorescein angiography was performed in two cases. Classification of the neoplasms was determined by fine-needle aspiration and biopsies of peripheral metastatic lesions, and/or complete necropsies. All four cases had unilateral or bilateral blindness and ophthalmoscopic lesions characterized by a wedge-shaped, tan discoloration in the tapetal fundus, variable but mild serous exudation under the retina, and profoundly attenuated retinal vasculature. Painful swelling and necrosis of the distal extremities and/or mass lesions in the appendicular musculature were also present. Clinical findings, along with microscopic findings from biopsy specimens or complete postmortem examination, documented widespread metastasis of variably differentiated, neoplastic, columnar epithelial cells presumed to be of bronchial origin. Tumor cells were predominately located within the systemic vasculature, consistent with classification of angioinvasive pulmonary carcinoma. Fluorescein angiographic and histopathologic findings in the affected globes suggested that the posterior segment lesions resulted from invasion and growth of neoplastic cells within the chorioretinal vasculature, resulting in secondary ischemic necrosis of the retina and choroid. Ischemic chorioretinopathy and necrosis of the distal extremities, associated with primary bronchogenic carcinoma, appear to be a unique neoplastic syndrome in the domestic cat. PMID- 11397255 TI - A dermoid of the eye in a blue-fronted Amazon parrot (Amazona aestiva). AB - A corneo-conjunctival dermoid is reported in a blue-fronted Amazon parrot (Amazona aestiva). After laminar keratectomy, histology showed the epidermis with feather follicles and dermal connective tissue with lymph follicles and sebaceous glands. PMID- 11397256 TI - Orbital neurofibrosarcoma in a dog. AB - A young dog was presented with rapidly progressive, unilateral, exophthalmos. Ultrasound-guided fine-needle aspiration of the retrobulbar mass resulted in a diagnosis of fibrosarcoma. Magnetic resonance imagery revealed tumor invasion into the brain, and palliative therapy was elected. The dog was euthanized 4 weeks following diagnosis due to progressive neurological signs. The final diagnosis was neurofibrosarcoma involving the pons, brainstem, left orbit and left trigeminal nerve. PMID- 11397257 TI - Viscoelastic materials in veterinary ophthalmology. AB - Viscoelastic materials have applications in both human and veterinary ophthalmology and orthopedics and for humans in otorhinolaryngology, urinary incontinence, cosmetic surgery and cosmetics. In ophthalmology, viscoelasties are most commonly used in facilitating cataract and other anterior segment surgeries, but also have applications in posterior segment surgery and in the topical management of keratoconjunctivitis sicca. Viscoelastics are routinely used during anterior segment surgery in filling and maintaining the anterior chamber, repositioning the iris, and to coat and protect the corneal endothelium, and expand the capsular bag prior to intraocular lens implantation. Viscoelastics can also be essential in the management of intraoperative complications such as miosis, hemorrhage, posterior capsular tears, and vitreous presentation. The most common products available for ophthalmic use include various concentrations of sodium hyaluronate, chondroitin sulfate, and hydroxypropylmethylcellulose. The physical properties of each viscoelastic material are dependent on chain length, and intra-and interchain interactions. The rheologic properties of viscosity, pseudoplasticity, viscoelasticity, and surface tension dictate the usefulness of each material for a specific purpose. PMID- 11397258 TI - Comparison of Optisol-GS and neomycin-polymyxin B-gramicidin ophthalmic solution for corneal storage in the dog. AB - The objective of the research was to compare the efficacy of Optisol-GS (OGS, Bausch & Lomb Surgical, Irvine, CA, USA) with triple antibiotic ophthalmic solution (neomycin-polymyxin B-gramicidin, NPG; Bausch & Lomb, Tampa, FL, USA) in preserving the viability of corneal endothelial cells. The study subjects were thirty young to middle-aged dogs with no gross corneal pathology that had been euthanized by pentobarbital overdose for reasons unrelated to this project. Corneal tissues were harvested, analyzed, and randomly assigned to treatment groups: one of two media (OGS or NPG), and one of five storage times (1, 7, 14, 21, or 35 days). Six corneas were stored in each medium for each time period. Corneal endothelial cell viability was evaluated pre- and poststorage by vital staining (trypan blue and alizarin red S), and endothelial cell morphology was evaluated with scanning electron microscopy. Storage in NPG caused significant loss (100%) of endothelial cells after all storage times. OGS storage maintained a high level of endothelial cell viability up to 21 days (98.9% +/- 1.3% viability). A significant decrease in percentage viability was also found for OGS stored corneas between 21 and 35 days, when endothelial cell viability decreased to 61.4% +/- 45.9%. The conclusions are that NPG storage at -20 degrees C is a very poor choice of media for corneal tissue banking if graft clarity is the goal. Storage in Optisol-GS at 4 degrees C for up to 21 days resulted in significantly higher percentages of viable endothelial cells. Optisol-GS storage should facilitate corneal preservation for canine keratoplasty patients. PMID- 11397259 TI - Immunohistochemical analysis of lens epithelial-derived membranes following cataract extraction in the dog. AB - The objective of the study was to characterize the morphologic and immunohistochemical features of lens epithelial-derived proliferative membranes from the anterior segment of canine globes. These features were correlated with those previously identified for diseases resulting from lens epithelial cell (LEC) proliferation including posterior capsular opacification, traumatic subcapsular cataract, and subcapsular plaques associated with hypermature cataracts. Sixteen canine globes were removed as a result of glaucoma or other complications following cataract extraction. Light microscopic and immunohistochemical analysis was performed on sections from formalin-fixed, paraffin-embedded globes. The tissues were stained with a variety of antibodies for cellular markers for LECs, growth factors or other cellular constituents relevant to cellular metaplasia and proliferation. The membranes were composed of monolayers or multilayers of spindle-shaped cells on the external surfaces of the anterior and posterior lens capsule, ciliary processes, iris leaflets, and iridocorneal angle, and they could be seen extending from an obvious monolayer of LEC within the capsular sac. Variably, scattered pigment cells, presumably of uveal origin, were concurrently present. Cellular components of the membranes stained positive for vimentin, transforming growth factor-beta, basic fibroblast growth factor, and smooth muscle actin. An amorphous eosinophilic extracellular matrix consisting predominately of collagen was associated with the membranes. Proliferative anterior segment membranes following cataract surgery were morphologically and immunohistochemically similar to cellular and matrix components of posterior capsular opacification and capsular plaques seen with hypermature cataracts, both of which result from metaplasia and proliferation of LEC. The presence of these LEC-derived membranes in association with secondary glaucoma suggests that exuberant proliferation of LEC outside the confines of the lens capsular sac may cause pathologic alterations in the eye following cataract surgery in the dog. PMID- 11397260 TI - A retrospective-cohort study on the development of cataracts in dogs with diabetes mellitus: 200 cases. AB - The objective of the study was to determine the incidence and estimated median time to cataract formation in dogs with diabetes mellitus. The animals studied were 200 dogs with diabetes mellitus which were referred to a university teaching hospital between 1985 and 1995. Medical records from dogs with a diagnosis of diabetes mellitus were reviewed and, where necessary, further follow-up information was gathered from the referring veterinarian. Incidence rate and median time to diabetic cataract formation was calculated using survival-analysis techniques in a retrospective cohort study design. Among the 200 dogs in the study population, 23 had cataracts at the time of diabetes diagnosis that were presumed to be related to other disease processes. Of the remaining 177 dogs, 132 had documented cataract development with features suggestive as being secondary to diabetes. Twenty-three dogs did not have obvious cataracts at the time of their last examination while 22 dogs did not have cataracts at the time they were lost to follow-up. These 55 cases contributed to the statistical models as noncases of cataracts until the last date for which an examination was available. Half of the population had developed cataracts by the 170th day postdiagnosis of diabetes mellitus, while 75% and 80% of the population developed cataracts by 370 days and 470 days, respectively. The results of this study suggest that the majority of dogs with diabetes will develop cataracts within 5-6 months from the time of diagnosis of the disease, and that approximately 80% of dogs will develop cataracts within 16 months of diagnosis. PMID- 11397261 TI - Tear production in dogs with epiphora and corneal epitheliopathy. AB - Measurement of tear volume by phenol red thread tear test (PRT), Schirmer's tear test (STT-1) and Schirmer's tear test with topical anesthesia (STT-2), and vital staining with a combination of fluorescein and rose Bengal of the cornea were performed in dogs with normal eyes and those with epiphora. The breeds included the Shih-Tzu (n = 26), and five other breeds (n = 50). The PRT, STT-1 and STT-2 results from the five breeds of normal dogs were 26.9 +/- 3.0 mm, 17.4 +/- 4.3 mm, and 8.8 +/- 3.2 mm, respectively. The PRT, STT-1, and STT-2 results for the Shih-Tzu eyes were (mean +/- standard deviation): 28.2 +/- 4.3 mm, 19.5 +/- 4.1 mm, and 9.2 +/- 4.5 mm, respectively. In the five breeds, corneal epitheliopathy, as evidenced by the retention of topical fluorescein and rose Bengal, occurred in 97% of dogs with epiphora and in 55% of the dogs without epiphora. Also in the dogs with corneal epitheliopathy the STT-2-value was low (4.0 +/- 2.8 mm) compared with those eyes without corneal epitheliopathy (8.8 +/- 3.1 mm). In the Shih-Tzu breed the STT-2 results were not significantly different between those dogs with corneal epitheliopathy and those with normal corneas. In the non-Shih Tzu breeds the decrease in basic tear secretion, as measured by the STT-2, is associated with the corneal epitheliopathy. PMID- 11397262 TI - Photoreceptor density of the domestic pig retina. AB - The spatial distribution and densities of photoreceptors in seven whole-mounted porcine retinas were studied and maps illustrating photoreceptor topography were constructed. Total photoreceptor densities ranged from to 83 000 to 200 000 cells/mm2, with a mean of 138 500 cells/mm2. Cone densities ranged from 39 000 (area centralis) to 8500 cones/mm2 (peripherally), with a mean of 16 400 cones/mm2. Rod:cone ratios ranged from 3:1 centrally to 16:1 peripherally, with a mean ratio of 8:1. Averaged photoreceptor densities are greatest (166 000 cells/mm2) within the central inferior retina, and regional differences in rod:cone ratios were found. Cone densities are increased in a broad region dorsal to the optic disk, extending both nasally and temporally. This region is believed to represent the area centralis. Cone densities gradually decrease and taper towards the periphery and inferior retina as rod:cone ratios increase. In addition to the many anatomic and ultrastructural similarities to the human eye, this study illustrates similarities within the photoreceptor mosaic of these two species and supports the use of the pig retina as a model for human/animal research. PMID- 11397263 TI - A matched observational study of canine survival with primary intraocular melanocytic neoplasia. AB - A retrospective histopathologic study was performed to evaluate the effect of primary intraocular melanocytic neoplasia on canine survival. Tumor size, location within the globe, extent of infiltration, and mitotic index were analyzed for their potential to predict survival. A total of 244 cases of dogs with melanocytic tumors submitted to the Comparative Ocular Pathology Laboratory of Wisconsin from 1988 to 1998 were evaluated. Histopathologic criteria (mitotic index, cytologic features of anaplasia) were used to differentiate 188 benign melanocytomas from 56 malignant melanomas. Signalment evaluation of age, sex, and breed revealed similarities in both tumor populations, with the majority of tumors discovered in 9-year-old, female/spayed, mixed-breed dogs. A greater percentage of left eyes (66%) vs. right eyes (47%) was found in the melanoma population, but an equal distribution was found in the melanocytoma population (48% and 52%, respectively). The majority of tumors arose from the anterior uveal tract (79% in the melanocytoma and 95% in the malignant melanoma populations). The German Shepherd breed was predisposed in the limbal distribution. At the time of enucleation, most tumors had invaded the sclera, but did not show extrascleral extension (51% in the melanocytoma and 61% in the malignant melanoma populations). Survival analysis showed a significant difference in survival between control and malignant melanoma populations (P = 0.0081) and was suggestive of a difference between the melanocytoma and melanoma populations (P = 0.031). Tumor extension, tumor size, and mitotic index were not found to be reliable predictors of survival. PMID- 11397264 TI - Bacterial and fungal pathogens isolated from corneal ulcerations in domesticated elephants (Elephas maximus maximus) in Sri Lanka. AB - Infective corneal diseases are common among the domesticated elephants in Sri Lanka, causing morbidity including blindness leading to loss of revenue to their owners. From 140 elephants of different ages and both sexes, 36 animals (25.7%) had evidence of keratitis, corneal ulcers, corneal opacities, and some had foreign bodies in their eyes. Nine elephants (6.4%) had lesions in both eyes (6.41%). Cultures for both bacteria and fungi were obtained from 26 corneal ulcers, including the nine elephants with bilateral lesions. The other 10 animals could not be restrained for sample collection. Swabs from the normal corneas of an additional 20 elephants without signs of any ophthalmic diseases were also collected. Twenty-three of the 35 (65.71%) samples from affected corneas yielded bacterial pathogens, and 14 (40%) also had fungal isolates. None of them yielded a fungal isolate alone. The predominant bacteria isolated were Staphylococcus aureus, beta hemolytic streptococci and coliforms. Fusarium, Cladosporium, Curvularia and Aspergillus species were the primary fungal isolates. No bacteria or filamentous fungi were isolated from the eyes with the normal corneas. Microbial identification including fungal isolations is suggested in the management of infective corneal diseases in elephants. PMID- 11397265 TI - Intraocular melanoma in an alpaca. AB - A case of primary intraocular melanoma in an alpaca is reported. This patient also presented with a mild normocytic, normochromic anemia and hypoproteinemia. These can be markers of an immunodeficiency syndrome associated with llamas. Although this patient did not appear to have clinical signs of this syndrome, its presence needed to be ruled out as animals affected with this syndrome are subject to recurrent severe debilitating diseases and death. To our knowledge, intraocular melanoma has not been previously reported in an alpaca. PMID- 11397266 TI - Ulcerative keratitis associated with qualitative tear film abnormalities in cats. AB - Three cats with indolent corneal ulcers and one cat with bilateral corneal sequestration and normal aqueous tear production were found to have rapid tear break-up times (BUTs). Tear BUTs in clinically affected cats averaged 2.5 +/- 1.29 s and 2.33 +/- 0.58 s for the right and left eyes, respectively. Palpebral conjunctival biopsies were harvested from consistent sites from each eye of affected cats (n = 7 affected eyes), and age-and breed-matched controls (n = 2 unaffected eyes). Light microscopy revealed a marked decrease to complete absence of conjunctival goblet cells (average goblet cell (GC):epithelial cell (EC) density = 18:50), conjunctival epithelial dysplasia, squamous metaplasia, and neutrophilic and mononuclear cell submucosal infiltration in affected cats. Specimens from the control cats had an average GC:EC density of 34:50, and minimal submucosal inflammatory infiltrate. The corneas (n = 7 eyes) healed following surgical keratectomy with (n = 2 eyes) or without (n = 1 eye) conjunctival pedicle flaps, superficial keratectomy and striate keratotomy with (n = 2 eyes) or without (n = 2 eyes) third eyelid flaps, and mucinomimetic tear supplementation (n = 5 eyes). Goblet cell regeneration was confirmed after 5 months of mucinomimetic supplementation (n = 2 eyes). The etiology for these mucin deficiencies remains unknown. PMID- 11397267 TI - Corneal stromal abscess in a horse. AB - A thoroughbred yearling presented with a focal, yellow, midstromal corneal opacity with concurrent iridocyclitis which was consistent with a corneal stromal abscess. When continued, appropriate, medical therapy failed to improve the patient's condition, penetrating keratoplasty was performed for diagnosis and therapy. Histopathology showed that the deep corneal stroma and Descemet's membrane were severely infiltrated with necrotic neutrophils and numerous, intralesional fungal hyphae. Culture was negative. Bacteria were not isolated, consistent with a fungal corneal stromal abscess. The resultant corneal scar did not interfere with the horse's racing career. PMID- 11397268 TI - Effect of topical atropine on intraocular pressure and pupil diameter in the normal horse eye. AB - The objective was to determine whether topically administered 1% atropine would alter intraocular pressure. The animals studied were four healthy adult horses. Intraocular pressure and pupil diameter were measured prior to and during a 2-day period of treatment with 1% atropine sulfate. No significant changes in intraocular pressure occurred as a result of the treatment with atropine. Pupil diameter increased significantly after atropine was applied. Available information on the outflow of aqueous humor from the horse eye suggests that atropine might reduce intraocular pressure in the horse by increasing uveoscleral outflow. This could prove beneficial in the treatment of equine glaucoma. We could not confirm a significant pressure-lowering effect of atropine. It is possible, however, that a longer treatment period may be required or that atropine may have a more profound effect on glaucomatous globes. PMID- 11397269 TI - Treatment of presumed iris melanoma in dogs by diode laser photocoagulation: 23 cases. AB - A semiconductor diode laser was used to cause remission of isolated presumed iris melanoma in 23 dogs. All cases presented as unilateral areas of raised iris hyperpigmentation, ranging in size from 2x 3 mm to 4x12 mm. Cases were treated using a diode laser delivery system through either an operating microscope adapter (OMA) or a laser indirect ophthalmoscope (LIO) with a 20D lens. Laser treatment was delivered 'to effect' using power ranging from 80 to 1000 mW and cumulative durations up to 14 min, 31 s (14:31). Immediate shrinkage of the mass was noted following treatment. Five cases required more than one laser treatment with three eyes receiving two treatments and two eyes receiving three treatments. Follow-up from the last laser treatment ranged from 6 months to 4.5 years during which time the lesions exhibited no enlargement. Minor complications related to laser treatment were seen, including: dyscoria, iris hyperpigmentation, and corneal edema due to collateral hyperthermia. Glaucoma and cataract formation were not observed. Non-invasive diode laser photocoagulation appears to be a safe and effective method of treatment for isolated, pigmented iris masses in dogs. PMID- 11397270 TI - p53 protein expression in conjunctival squamous cell carcinomas of domestic animals. AB - The expression of p53 protein was investigated in eight formalin-fixed, paraffin embedded conjunctival squamous cell carcinomas of five horses and one cow, dog and cat each by an immunohistochemical procedure in order to evaluate protein overexpression. Anti-human p53 protein mouse monoclonal antibodies known to be cross-reactive with p53 protein of the animal species examined were used. Positive p53 nuclear immunostaining was detected in five equine, one bovine and one feline cases. Conversely, no p53 immunostaining was found in the only canine case examined. These results demonstrate a frequent p53 overexpression in conjunctival squamous cell carcinoma that could be related to UV-induced mutations of the p53 tumor suppressor gene. PMID- 11397271 TI - A retrospective study of 30 cases of frozen lamellar corneal graft in dogs and cats. AB - Frozen lamellar corneal grafts and nictitating membrane flaps were used in 18 dogs and 12 cats to repair deep corneal defects. In all dogs either melting corneal ulcers or descemetoceles were present. In the 12 cats, nine had either a melting corneal ulcer or descemetocele, two animals had acute bullous keratopathy, and one cat had corneal sequestrum. Initial vascularization with gradual clearing of the graft occurred during the first 45 days postoperatively. At 60 days postoperatively, all eyes were visual. Frequent postoperative complications included: focal dehiscence of the wound (n = 9); melting of part of the graft (n = 7); and pigmentation of the graft (n = 4). The frozen lamellar corneal graft was a very safe technique, and restored the tectonic and the optical function of the cornea. It provided the best results in corneas with nonperforating corneal defects. This technique provides poorer results when the cornea was perforated prior to surgery or during the surgical procedure. PMID- 11397272 TI - Treatment of equine glaucoma by transscleral neodymium:yttrium aluminum garnet laser cyclophotocoagulation: a retrospective study of 23 eyes of 16 horses. AB - Contact neodymium:yttrium aluminum garnet (Nd:YAG) laser transscleral cyclophotocoagulation (TSCP) was performed on 23 eyes of 16 horses for treatment of glaucoma. The mean highest preoperative IOP was 51 +/- 17 mmHg. Follow-up evaluation was available for 19 eyes 1 day after surgery, 14 eyes from 1 to 2 weeks, 16 eyes from 4 to 6 weeks, 9 eyes from 12 to 16 weeks, and 10 eyes greater than 20 weeks after laser treatment. The mean intraocular pressure (IOP) the day following surgery was 34 +/- 13 mmHg. The mean IOP for each follow-up period was: one to two weeks postoperative, 23 +/- 9 mmHg; four to six weeks, 24 +/- 7 mmHg; 12-16 weeks, 28 +/- 10 mmHg; and >/= 20 weeks, 22 +/- 9 mmHg. IOP measurements were significantly different from pretreatment values for all follow-up intervals except for weeks 12-16 (P < 0.05). Treatment success was defined as maintenance of IOP < 30 mmHg. Treatment success was achieved in 93%, 88%, 78%, and 70% of the treated eyes at the 1-2 weeks, 4-6 weeks, 12-16 weeks, and the >/= 20 weeks re evaluation, respectively. No significant difference was found between the number of eyes visual at presentation (52.2%) and visual at 20 weeks (60%). The most common laser complications were conjunctival hyperemia (21.7%) and corneal ulcers (13.0%). Results of this study indicate that Nd:YAG TSCP is an effective method of controlling IOP and preserving vision in horses with glaucoma. An effective Nd:YAG laser protocol for TSCP in the equine glaucomatous eye is a power setting of 11 W, duration of 0.4 s, applied 5 mm posterior to the limbus at 60 sites, resulting in a total energy dose of 264 J. PMID- 11397274 TI - Editorial. PMID- 11397273 TI - Spontaneous buphthalmos in the Djungarian hamster (Phodopus sungorus campbelli). AB - Four Djungarian hamsters (Phodopus sungorus campbelli) were examined because of insidious globe enlargement, visual impairment and secondary altered behavior. Buphthalmos, deep anterior chambers, widely dilated pupils and poor to absent pupillary light reflexes were evident bilaterally in all four animals. Pale retinae and atrophic optic nerve heads were observed in the oldest hamsters. Light microscopic findings, including uveal atrophy, retinal atrophy (especially the inner retina), and a relative decrease in ganglion cells and optic nerve atrophy, were consistent with glaucoma. The etiology was unclear, but a genetic predisposition cannot be excluded. PMID- 11397275 TI - Unilateral external and internal ophthalmoplegia caused by intracranial meningioma in a dog. AB - Unilateral internal and external ophthalmoplegia caused by an intracranial meningioma occurred in a 15-year-old Belgian Sheepdog. The dog initially presented with ventro-lateral strabismus of the left eye, and ptosis of the left upper eyelid. Anisocoria was present with the left pupil fixed and dilated. Both eyes were visual. Neuro-ophthalmic evaluation revealed a lesion located in the left oculomotor nerve. Pharmacological testing with dilute pilocarpine (0.1% in artificial tears) revealed evidence of parasympathetic denervation of the left eye. Further evaluation via magnetic resonance imaging (MRI) revealed a well defined mass to the left of midline and lateral to the sella turcica. An attempt was made to excise/debulk the mass due to worsening conditions and the dog died the following day. Necropsy revealed a mass of randomly arranged bundles and streams of spindle cells. Immunohistochemistry demonstrated a strong avidity for vimentin and a negative response for S-100 protein. These findings suggest a diagnosis of meningioma. PMID- 11397276 TI - Canine orbital meningiomas: a review of 22 cases. AB - Clinical and pathologic features of primary orbital meningiomas in the dog were reviewed. Twenty-two meningiomas, confined to the orbit, were identified from the Comparative Ophthalmic Pathology Laboratory of Wisconsin from 1981 to 1997. The dogs ranged in age from 3 to 17 years (mean = 9.2 years). The clinical presentation, reported in 20 cases, was indicative of a retrobulbar mass and included exophthalmos and orbital swelling (18/20), and papilledema or abnormalities of the posterior segment (7/20). Visual acuity was reported in 15 cases; of those, 12 dogs were blind in the affected eye. Follow-up information was obtained on 17 cases; six dogs developed local recurrence of the neoplasm. Two dogs with recurrent neoplasms simultaneously developed blindness in the opposite eye. Extension along the optic nerve to the optic chiasm was suspected. No metastasis was found at the time of the study. Enucleation with excisional biopsy was effective therapy to date in 11 cases (0.2-4.5 years follow-up time). All neoplasms were located within the vicinity of the optic nerve and, when sectioned through the optic nerve head, appeared to completely envelope the nerve. The neoplastic cells were arranged in tight whorls and bundles characteristic of meningiomas. Most tumors had islands of chondroid and osseous metaplasia (17/22). Ocular invasion was limited to small foci in the posterior choroid or optic nerve head of six dogs. Immunoperoxidase stains on 10 cases were positive for vimentin and S-100, but negative for cytokeratin. Electron microscopy revealed complex interdigitations between cell membranes and few desmosomal intercellular attachments. Primary orbital meningiomas have a characteristic histologic appearance and may recur locally after surgical excision. PMID- 11397277 TI - Evaluation of intraocular and partial CO2 pressure in dogs anesthetized with propofol. AB - The effects of propofol on intraocular pressure (IOP) and end tidal CO2 (ETCO2) were studied because an elevation in the latter may alter IOP. Twenty dogs were divided into two groups (G1 and G2). G1 dogs were induced with 10 mg/kg (IV) of propofol followed by a 0.4 mg/kg/min continuous infusion of the same agent diluted in a 0.2% dextrose solution for 1 h. G(CAPS) 2 dogs served as the control group, where only dextrose solution was administered, under the same time intervals as in G1. Applanation tonometry (Tono-Pen) was used to determine IOP and ETCO2 as a method to determine partial CO2 pressure. Measurements were taken every 15 min for 1 h, with M1 occurring immediately before IV administration. IOP and ETCO2 were not statistically significant in either groups. Based on the results, it may be concluded that propofol does not alter IOP and ETCO2. PMID- 11397278 TI - Extraocular muscle myositis and restrictive strabismus in 10 dogs. AB - Ten cases of uni- or bilateral restrictive ventromedial strabismus in young dogs of different breeds are reported. Clinically, abnormalities were restricted to the extraocular muscles with sparing of the masticatory muscles and limb muscles. This was supported in some cases by imaging studies, electrophysiology, and immunocytochemical assay for antibodies against type 2M fibers. Histologically, there was variable lymphocytic plasmacytic mononuclear cell infiltration and fibrosis. This disorder is similar in many aspects to chronic masticatory myositis with focal myositis and subsequent fibrosis. Surgical correction may restore eye position and vision. PMID- 11397279 TI - Congenital scleral staphyloma in a dog repaired with preserved homologous peritoneum. AB - A case of congenital scleral staphyloma was treated with a preserved homologous peritoneum graft. A 2-year-old female poodle with a blue-gray mass in a dorsal and temporal position of the sclera of the left eye was evaluated. The repair of the scleral defect with a graft of preserved homologous peritoneum reduced the strabismus and the size of the staphyloma, and improved the cosmetic appearance of the eye. PMID- 11397280 TI - Intraocular pressure in normal llamas (Lama glama) and alpacas (Lama pacos). AB - OBJECTIVE: To determine the mean intraocular pressure in llamas (Lama glama) and alpacas (Lama pacos) using applanation tonometry.Animals studied: Ten llamas and 10 alpacas.Procedures: Intraocular pressure (IOP) was measured with a Tono Pentrade mark XL (Mentor Ophthalmics, Inc., Norwell, MA, USA). Three values, with 5% variance, were recorded for each eye. Least-squares means were determined for IOP for each eye of llamas and alpacas. Controlling for age, differences between left and right eye were analyzed using ANOVA. Two age groups were established, less than 5 years and greater than 5 years. The effect of age on IOP within each group was analyzed by linear regression. Probability values of less than 0.05 were considered significant. RESULTS: Comparison of mean IOP between right (n = 20) and left eyes (n = 20), independent of species type, showed no differences in IOPs for llamas and alpacas. Mean IOP declined with increasing age in llamas and alpacas. Mean IOPs for 20 eyes in 10 llamas was 16.96 +/- 3.51 mmHg. Mean IOP for 20 eyes in 10 alpacas was 16.14 +/- 3.74 mmHg. Mean IOP for all eyes (n = 40), independent of species, was 16.55 +/- 3.55 mmHg. The range of IOP in normal llamas and alpacas within 2 SD (95% of the population) was 14.89+/-18.21 mmHg. CONCLUSIONS: There was no significant difference in IOP between alpacas and llamas. Mean IOP in both species decreased with increased age. PMID- 11397281 TI - Ocular and periocular manifestations of leishmaniasis in dogs: 105 cases (1993 1998). AB - The purpose of this retrospective study was to determine the prevalence, type, and prognosis of ocular lesions associated with leishmaniasis in dogs. One hundred and five dogs (24.4% of all cases of leishmaniasis diagnosed during the study period) had ocular or periocular leishmaniasis, and 16 dogs (15.2% of ocular cases) had only ocular lesions and systemic signs were not apparent. Anterior uveitis was the most common manifestation and other prevalent findings included blepharitis and keratoconjunctivitis. Several distinct variations of eyelid lesions were seen including a dry dermatitis with alopecia, diffuse blepharedema, cutaneous ulceration, and discrete nodular granuloma formation. In some cases with keratoconjunctivitis, corneal lesions clinically resembled nodular granulomatous episclerokeratitis. Twenty-seven of the 34 cases with ocular lesions had improvement in signs following systemic antiprotozoal and topical anti-inflammatory therapy, although many cases with anterior uveitis required long-term topical therapy. Response of ocular signs correlated highly with overall, systemic response to therapy. Ophthalmic manifestations of systemic leishmaniasis are common in the dog, and this disease should be considered in the differential diagnosis of most adnexal and anterior segment ocular inflammatory lesions in dogs in endemic areas. PMID- 11397282 TI - Immunohistochemical characterization of intraocular metastasis of a canine transmissible venereal tumor. AB - Little has been published on intraocular metastasis of transmissible venereal tumors (TVT) in dogs. This report presents a 4-year-old male Labrador Retriever with a previous history of subcutaneous TVT which underwent total remission after treatment with vincristine. The dog presented with clinical signs of uveitis and increased intraocular pressure (IOP) in both eyes. After enucleation of the left eye, a diagnosis of TVT was made based on morphology, histology and immunohistochemistry (IHC). IHC staining for vimentin, S-100 protein, cytokeratin and HMB45 was performed to differentiate this lesion from TVT, lymphoma, melanoma, carcinomas, neurogenic tumors and fibrosarcoma. The IHC findings supported the diagnosis of TVT for this round cell tumor. PMID- 11397283 TI - Editorial. PMID- 11397284 TI - Equine conjunctival pseudotumors. AB - Five horses presented with unilateral pink, smooth, nonulcerated conjunctival masses with histologic features characteristic of inflammatory pseudotumors, i.e. proliferative inflammatory lesions clinically resembling true neoplasia. Although causes for the inflammatory lesions were not determined, based on the presence histologically of mononuclear (predominantly lymphocytic) inflammatory cell infiltrates and the absence of infectious agents, parasites or foreign bodies, an immune-mediated pathogenesis was suspected. Affected horses ranged from 5 to 8 years of age with no apparent breed or sex predilection. Conjunctival lesions were nodular in two cases and relatively flat and more diffuse in three cases. Third eyelid lesions were present in three cases and two affected eyes had corneal involvement. Based on findings from these five cases, the prognosis for equine conjunctival pseudotumors appears to be good when lesions are treated by partial or complete surgical excision, local administration of anti-inflammatory agents, or a combination of surgery and anti-inflammatory therapy. PMID- 11397285 TI - Lens opacities in the horse: a clinical classification. AB - Lens opacities, including cataracts, are frequent findings during routine ophthalmic examination in the horse, and clinicians often experience difficulty in predicting the potential progression and consequences on visual function of these opacities. This has particular relevance to the prepurchase examination. The present study proposes a descriptive classification of the more common lens opacities observed in the horse based principally on their morphological origin. This classification provides a basis for objective prediction of the probable progression of lens opacities, in particular cataracts, in the horse. PMID- 11397286 TI - Disease of the equine posterior segment. AB - The appearance of the equine fundus is reviewed from the perspective of differentiating normal variations from disease, and the descriptions have been updated to include recently published ocular fundic abnormalities. Most pathological lesions are identified near the optic nerve head, and typically involve depigmentation or hyperpigmentation. Depending upon configuration and appearance, linear pigmented bands may reflect the course of the vortex veins, the transition from tapetal to nontapetal fundus, or indicate chorioretinitis or equine motor neuron disease. Choroidal vasculature is readily apparent in color dilute (subalbinotic) horses and must be differentiated from hemorrhage. Retinal hemorrhages in foals are common and may occur independently to hypoxic ischemic encephalopathy. Retinal cysts may signal more significant disease in the eye such as anterior segment dysgenesis. Prominence of gray or tan-colored material on or near the optic nerve head may represent traumatic optic neuropathy, benign optic neuropathy, proliferative optic neuropathy or actual neoplasia. PMID- 11397287 TI - Rose bengal positive epithelial microerosions as a manifestation of equine keratomycosis. AB - Purpose To describe the clinical appearance of corneal epithelial cell microerosions associated with keratomycosis in the horse. METHODS: Retrospective clinical study. RESULTS: Multifocal, punctate, superficial corneal opacities with positive rose bengal retention were noted in six horses with presumed 'viral keratitis'. Faint fluorescein staining was also present in three cases. Equine herpesvirus tissue culture inoculation was negative for a cytopathic effect in three cases. Aspergillus (n = 3), Curvularia (n = 1), and an unidentified fungus (n = 1) were cultured in five horses, and hyphae found on corneal cytology from the sixth. Mixed bacterial infections were present in three eyes. The eyes of two horses with Aspergillus progressed to deep melting corneal ulcers that required surgical therapy. The microerosions remained superficial, but persistent in the other four eyes. Natamycin was utilized topically in all six horses. Transmission electron microscopy from case 6 revealed mucin layer disruption, an intact corneal epithelial cell layer, and fungal attachment to degenerating epithelial cells. The visual outcome was positive in all six horses, although healing was prolonged (48.5 +/- 14.5 days on average in the horses with no surgery; 62 days on average in the two horses that required surgery). CONCLUSIONS: Complete removal or full-thickness penetration of the corneal epithelial cell barrier may not be necessary to allow fungal adherence and initiation of keratomycosis in the horse. Prior to colonization and invasion of the horse cornea, fungi may induce changes in the mucin layer of the tear film that result in or are associated with rose bengal positive microerosions of the superficial corneal epithelium. Horses with painful eyes, and eyes with superficial, multifocal corneal opacities should have their corneas stained with both fluorescein and rose bengal as fungal microerosions may stain weakly, or not at all, with fluorescein, and may thus be mistaken for presumed 'viral keratitis' of the horse. PMID- 11397288 TI - Tear-deficient and evaporative dry eye syndromes of the horse. AB - Tear-deficient dry eye (keratoconjunctivitis sicca), and evaporative dry eye, with some overlap between these two categories, are two major categories of dry eye recognized in the horse. Careful examination of the eyelid margins, the inner and outer surfaces of the upper and lower eyelids, assessment of blink and third eyelid movement, specific testing of tear production, assessment of corneal sensitivity, and slit-lamp examination of the ocular surface before and after fluorescein dye application is recommended to detect dry eye problems. Rose Bengal dye application is also recommended in many cases. Facial nerve paresis is a possible complicating factor in some instances. Evaporative dry eye deserves closer scrutiny for it may be a contributory factor when ocular surface disease such as keratomycosis is present. Factors which influence ocular surface health need more close examination in horses. PMID- 11397289 TI - Developmental anomalies and abnormalities of the equine iris. AB - The iris is derived from interaction between neural crest tissue and the adjacent neuroectoderm of the optic cup. Developmental anomalies of the equine iris are common, and include congenital miosis, iris cysts, various manifestations of iris hypoplasia, heterochromia, and persistent pupillary membrane remnants. They may be found alone or in combination with other ocular defects. PMID- 11397290 TI - Posterior lamellar keratoplasty for treatment of deep stromal absesses in nine horses. AB - OBJECTIVE: To describe and evaluate the use of posterior lamellar keratoplasty as a surgical treatment for deep corneal stromal abscesses in horses. Animals studied Nine horses of various breeds and ages that presented with corneal stromal abscesses located in the posterior one-third of the cornea. Procedure Retrospective medical record study. RESULTS: Nine horses had deep corneal stromal abscesses that were treated with posterior lamellar keratoplasty. Median patient age was 3 years. Six patients were females and three were geldings. Medical therapy alone had been attempted prior to surgery in all nine animals. Corneal abscess culture and histopathology were performed in 8/9 horses. Cultures were positive for an infectious etiology in 4/8 (50%). Histopathology was positive for an infectious etiology in 5/8 (62.5%). Mean surgical time was 71.0 +/- 18.8 min and the average healing time was 23.7 +/- 5.2 days. Visual outcome was positive in 8/9 cases. Conclusion Posterior lamellar keratoplasty is a promising procedure for treatment of deep corneal stromal abscesses in horses. The procedure resulted in considerable shorter surgery time and healing time than had been observed with full-thickness penetrating keratoplasty. Scar formation with this procedure was not significantly different than with penetrating keratoplasty. PMID- 11397291 TI - Long-term effect on the equine eye of an intravitreal device used for sustained release of cyclosporine A. AB - OBJECTIVE: To determine the long-term toxicity of an intravitreal device releasing continuous cyclosporinee A (CsA) in normal eyes of horses by evaluating clinical signs, electroretinography, and histopathology. Animals Studied Ten adult horses with normal ophthalmic examinations were used in this study Procedure(s) Four horses had one eye implanted with a CsA device, and six horses had the right eye implanted with a CsA-containing device (10 eyes with CsA in total) and the left eye (six eyes in total) with the device without drug (control). The implants were placed in the vitreous of the eyes through a sclerotomy 1 cm posterior to the limbus in the dorso-temporal quadrant of the eye. Scotopic electroretinograms were performed prior to implantation and at 1 week, and at 1, 3, 6, 9, and 12 months postimplantation. Two of the unilaterally implanted horses were euthanized at 1 weeks postimplantation, and two at 6 weeks postimplantation. Two of the bilaterally implanted horses were euthanized at 6 months, two at 9 months, and two at 12 months postimplantation. At euthanasia, the eyes were removed, aqueous and vitreous humor aspirated, and tissues fixed in 10% buffered formalin and processed for histopathology. CsA concentrations were measured by high pressure liquid chromatography in the aqueous and vitreous humors, and in peripheral blood. RESULTS: The devices were tolerated well in 14 of 16 eyes. There was minimal postoperative inflammation in most eyes, with a normal appearance within 7 days. In two eyes implanted with the CsA device, severe inflammation resulted in phthisis bulbi by 28 days. One of these eyes exhibited suspected bacterial endophthalmitis, and one had a sterile endophthalmitis and cataract presumably from trauma to the lens during implantation. In the other 14 eyes, no change was observed in the scotopic electroretinograms (ERG) from preoperative results, and no significant differences between the right (CsA) and left (control device) eyes were observed. CsA levels in the aqueous and vitreous humor, and peripheral blood were below the detection limit of the HPLC. Histologic findings revealed only a mild lymphoplasmacytic cellular infiltrate in the ciliary body and pars plana near the implantation site. CONCLUSIONS: The CsA devices were well tolerated with no long term complications from the implants themselves. However, complications may occur from inadvertent implantation trauma or contamination during surgery. The long term safety of the device may make it useful for delivery of CsA in the control of equine recurrent uveitis. PMID- 11397292 TI - Evaluation of tear film proteinases in horses with ulcerative keratitis. AB - Ulcerative keratitis is a common and potentially blinding ocular disease of horses, capable of progressing to corneal perforation in as little as 24 h. This rapid stromal degeneration is mediated in part by exogenous and endogenous proteinases. We measured and compared the concentrations of two matrix metalloproteinases (MMP-2 and MMP-9) and a serine proteinase (neutrophil elastase) present in the precorneal tear film of normal horses and horses with rapidly progressing ulcerative keratitis. Precorneal tear film samples were collected from 23 ulcerated and 21 unaffected eyes of 23 horses with unilateral ulcerative keratitis, and from 33 normal eyes of 17 control horses. MMP-2, MMP-9, and neutrophil elastase were identified by casein and gelatin zymography and quantified by computerized image analysis. Median MMP-9 levels were significantly higher in the precorneal tear film of young control horses vs. older control horses (P = 0.005). Median MMP-2, MMP-9, and neutrophil elastase levels were significantly higher in the precorneal tear film of ulcerated eyes when compared to age-matched normal controls (P = 0.004, P = 0.001, and P = 0.012, respectively). Median MMP-2 levels were also significantly higher in the precorneal tear film of contralateral eyes of affected horses when compared to age-matched normal controls (P = 0.004). No significant differences in median proteinase levels were detected between 'sterile' ulcers and those from which bacteria or mixed infections (bacteria and fungi) were isolated. However, median MMP-2 and neutrophil elastase levels were significantly higher in the precorneal tear film of eyes with 'sterile' ulcers when compared with ulcerated eyes from which fungi were isolated (P < 0.05). The results of this study support the use of topical antiproteinase therapy which targets both MMPs and serine proteinases in progressive equine ulcerative keratitis. PMID- 11397293 TI - Ulcerative keratitis caused by beta-hemolytic Streptococcus equi in 11 horses. AB - Purpose To describe 11 clinical cases of ulcerative keratitis in horses associated with beta-hemolytic Streptococcus equi in Florida, USA. METHODS: Retrospective clinical study (1996-99). RESULTS: Beta-hemolytic Streptococcus equi was cultured from 11 horses with deep ulcers, descemetoceles or iris prolapse (n = 8), a suture abscess found with a penetrating keratoplasty for a stromal abscess (n = 1), and ulceration that developed following keratectomy/irradiation for corneal squamous cell carcinoma (n = 2). Beta hemolytic Streptococcus equi subspecies zooepidemicus was found in 10 eyes and subspecies equi in one. Marked signs of uveitis including miosis and hypopyon were present in 8/11 (72.7%) eyes. Keratomalacia was severe in all eyes. The mean diameter of the ulcers associated with beta-hemolytic Streptococcus was 10.2 +/- 6.1 mm. Eight of the eyes required conjunctival flap surgery (four grafts dehisced) and one eye corneal transplantation. Two eyes were treated with medication only. Isolate sensitivity to antibiotics included ampicillin (6/11), bacitracin (11/11), cephalothin (11/11), chloramphenicol (11/11), gentamicin (5/11), polymyxin B (2/11), and tobramycin (1/11). All isolates were resistant to neomycin. The average healing time was 44.7 +/- 26.7 days. The visual outcome was positive in 8/11 eyes, and the globe retained in 9/11 eyes. CONCLUSIONS: Although Gram-positive bacteria predominate in the normal conjunctival microflora of horses throughout the world, Gram-negative bacteria and fungi are more often isolated from equine ulcers. Beta-hemolytic Streptococcus spp. are associated with a very aggressive ulcerative keratitis with the capability to digest conjunctival graft tissue. Clinical signs are pronounced. Aggressive surgical and intensive medical therapy with topical antibiotics and protease inhibitors is indicated. PMID- 11397294 TI - Immunohistochemistry of the extracellular matrix of the normal equine lamina cribrosa. AB - Purpose To use immunohistochemical techniques to identify and localize the structural macromolecules of the extracellular matrix (ECM) of the normal adult equine lamina cribrosa in order to make comparisons to the extracellular matrix of the lamina cribrosa of horses with glaucoma. METHODS: Normal eyes of five adult horses between 5 and 10 years of age were fixed in 10% neutral buffered formalin and embedded in paraffin. Polyclonal rabbit-derived antibodies against human elastin, laminin, fibrillin-1, and collagen types I, III and IV, and polyclonal goat-derived antibodies against collagen type VI were used as primary antibodies. Transverse and longitudinal histologic sections of the optic nerve head and lamina cribrosa were stained using several dilutions of the primary antibodies, biotinylated link antibody, horseradish peroxidase-labeled streptavidin, and 3,3'-diaminobenzidine as a chromogen. The immunohistochemical staining patterns were qualitatively interpreted. RESULTS: The normal adult horse lamina cribrosa labeled positively for collagen types I, III and VI, laminin, elastin and fibrillin. Collagen type VI staining of the laminar ECM was most intense, followed by labeling for collagen types III and I, respectively. Laminar blood vessels were weakly positive for laminin and slightly positive for type IV collagen. The scleral ECM of the laminar insertion zone had more intense labeling for collagen types I and VI than did the laminar plates. CONCLUSIONS: The extracellular matrix of the laminar plates of the adult equine lamina cribrosa is similar to the dog as it consists of elastic and collagen fibers (with collagen types VI, III and I). Both the normal dog and horse lamina display more intense staining of collagen type VI than is found in the ECM of the normal human lamina cribrosa. The macromolecular structure of the equine lamina cribrosa suggests that it is a very resilient structure that may provide some protection to the optic nerve axons during episodes of elevated intraocular pressure. PMID- 11397295 TI - Cochet-Bonnet aesthesiometer-determined corneal sensitivity in neonatal foals and adult horses. AB - Corneal touch threshold (CTT) was measured in sick neonatal foals, healthy foals, and healthy adult horses with a Cochet-Bonnet aesthesiometer. The mean overall CTT for the adult horses, sick foals, and healthy foals was 4.82 +/- 0.87 cm, 3.21 +/- 0.24 cm, and 5.01 +/- 0.61 cm, respectively. The central cornea of adult horses was more sensitive than the limbal cornea. Corneal sensitivity was significantly reduced in sick neonatal foals compared to adults. The mean Schirmer I tear test values were significantly lower in foals than adults, and were 14.2 +/- 1.0 mm, 12.8 +/- 2.4 mm, and 18.3 +/- 2.1 mm wetting in sick neonatal foals, normal neonatal foals, and adult horses, respectively. Reduced corneal sensation and lower tear production may be associated with ulcerative keratitis and slow corneal healing in some foals. PMID- 11397296 TI - Effect of topical 1% atropine sulfate on intraocular pressure in normal horses. AB - OBJECTIVE: To determine the effect of topical 1% ophthalmic atropine sulfate on intraocular pressure (IOP) in ocular normotensive horses. Animals Studied Eleven clinically healthy horses. Procedures IOP was measured bilaterally twice daily, at 8 AM and 4 PM, for 5 days. No medication was applied for the first 2 days of the study. Thereafter, one eye of each horse was treated with 0.1 mL of topical 1% atropine sulfate ointment twice daily (7 AM and 7 PM) for 3 days. The contralateral eye served as a control. In eight of the horses, an additional IOP reading was taken 3 days following cessation of the atropine treatment. RESULTS: There was no significant difference in the IOP of control vs. treatment eyes in the pretreatment period, days 1 and 2 (P = 0.97 and 0.55, respectively). During the treatment period, treated eyes of 10 of the horses had significantly lower IOP than control eyes (P = 0.03). The mean IOP reduction in treated eyes, relative to untreated eyes, was 11.2%. One horse had a significant rise in IOP in the treated eye compared to the remaining study animals. The IOP of control eyes did not vary significantly over the observation period (P = 0.27). There was no significant variation in IOP between the 8 AM and 4 PM measurement (P = 0.9). CONCLUSIONS: Topical 1% atropine sulfate causes a small, but significant decline in IOP in most ocular normotensive horses. Because topical atropine may elevate IOP in some horses, it should be used with caution in the treatment of glaucoma in this species. PMID- 11397297 TI - In vitro analysis of antiangiogenic activity of fungi isolated from clinical cases of equine keratomycosis. AB - OBJECTIVE: The goal of this project was to explore the possibility that fungal organisms produce metabolites that inhibit angiogenesis. Procedures Fungal cultures were obtained from cases of keratomycosis, grown in Sabouraud's dextrose broth, and sterile filtered for use in experiments. The Matrigel assay was used to screen the filtrate samples for antiangiogenic activity. Matrigel is a basement membrane matrix that supports the differentiation of human umbilical vein endothelial (HUVE) cells into a capillary-like network of tubules. HUVE cells were cultured using standard techniques and passaged at confluence, with all cells being used at passage 3-6. HUVE cells (40 000 cells) were pipetted into each well of a 24-well tissue-culture plate coated with Matrigel. An aliquot of fungal media filtrate was added to each well and the plates allowed to incubate for 18 h, at which time they were evaluated for tubule formation. RESULTS: Two fungal isolates showed inhibition of tubule formation. The addition of 100, 200 and 400 &mgr;L of the fungal media filtrate from the first isolate (Fusarium sp. 99A34574) produced a consistent and dose-dependent inhibition of tubule formation. The second isolate (Aspergillus sp. 271599) did not show inhibition of tubule formation with 100 or 200 &mgr;L added to the wells, however, it did show inhibition at 400 &mgr;L/well. The remaining three isolates did not cause inhibition at any concentration. CONCLUSIONS: Our findings suggest that certain fungal organisms produce metabolites that inhibit tubule formation in vitro, and that these metabolites may play a significant role in altering the host vascular response to fungal infections of the cornea. PMID- 11397298 TI - Inferomedial placement of a single-entry subpalpebral lavage tube for treatment of equine eye disease. AB - The objective of this study was to describe method of placement, and frequency and severity of complications associated with a subpalpebral lavage system placed in the medial aspect of the equine inferior eyelid. The inferomedial subpalpebral lavage (ISPL) tube is positioned deep in the medial aspect of the inferior conjunctival fornix so that the footplate lies flat between the lower eyelid and the anterior surface of the nictitans. Retrospective data from the placement of 92 ISPL systems placed in 86 horses during a 31-month period were examined. Tube placement was performed using sedation and regional anesthesia only in 59% of horses. The median duration of tube placement was 19 days (range: 1-61 days). Seventy-one horses were treated for up to 55 days following discharge from hospital with an ISPL tube in place. No complications were reported with 59% of ISPL systems. Non-ocular complications were found in 38% of ISPL systems and included tube displacement from the conjunctival fornix (18%), suture loss requiring resuturing of the system to the horse's head (14%), and damage necessitating replacement of the injection port (6%). Ocular complications were recorded in 3% of horses and were limited to inferior eyelid swelling. Vision was retained in 88% of horses. The ISPL system is easily and safely placed, and well tolerated for extended periods. It appears to be associated with infrequent and minor complications when compared with placement of subpalpebral lavage tubes in the superior eyelid. PMID- 11397299 TI - Cataract phacofragmentation in horses. AB - Cataract surgeries were carried out in fifty-one eyes of 36 horses over a 15-year period. Cataracts were removed using phacofragmentation and aspiration. Useful vision was restored after surgery in 30 horses. One year after surgery 16 of the 19 horses for which follow up information was available were still visual with several still being used as working horses. At 5-6 years after surgery three horses were still visual. The most frequent intraoperative complication was tearing of the posterior lens capsule. The most frequent postoperative problem was superficial corneal ulceration. Four eyes in three horses developed postoperative infectious endophthalmitis resulting in blindness. PMID- 11397300 TI - Effect of single- and multiple-dose 0.5% timolol maleate on intraocular pressure and pupil size in female horses. AB - OBJECTIVE: To determine the effect of single and multiple-dose 0.5% timolol maleate on intraocular pressure (IOP) and pupil size between 8 AM and 8 PM. Animals Nine female horses with normotensive eyes. Procedure IOP, horizontal and vertical pupil size were measured on a single day, between 8 AM and 8 PM at hours 0, 0.5, 1, 2, 4, 6, 8, 10, and 12. A single dose of 0.5% timolol maleate was applied to both eyes immediately after the first measurement at 8 AM. IOP and pupil size were measured at 8 AM and 4 PM in a 5-day experiment of twice-daily application of 0.5% timolol maleate. RESULTS: A significant decrease in IOP from 24.9 +/- 4.2 mmHg prior to application of timolol maleate to 20.7 +/- 3.1 mmHg (4.2 mmHg = 17%) was observed 8 h after single-dose application. A significant decrease in horizontal pupil size (2.0 mm = 11%) was present 6 h after single dose application. In the multiple-dose experiment, a significant decrease in IOP was present on days 4 and 5 as compared to IOP measured prior to application of timolol maleate. A significant decrease in horizontal and vertical pupil size was present throughout the 5-day study as compared to the values obtained prior to treatment. CONCLUSIONS: 0.5% timolol maleate significantly decreased IOP and pupil size in normo-tensive eyes of this group of female horses in both single and multiple twice daily applications. PMID- 11397301 TI - The diagnosis and treatment of periorbital sarcoid in the horse: 445 cases from 1974 to 1999. AB - OBJECTIVE: Diagnosis and management of periorbital sarcoids in horses is a significant clinical challenge for the practicing veterinary surgeon and pathologist. The purpose of this study was to investigate the response of various types of sarcoids to different therapeutic methods. Animals studied Medical records of 445 clinical patients. Procedures This paper retrospectively examines the clinical and histological features of periorbital sarcoids and the treatment of 445 cases. Treatment by surgical excision, cryosurgery, Bacillus-Calmette Gaerin (BCG) immunomodulation, topical cytotoxic applications, and radiation are described. RESULTS: Six types of sarcoid can be found in the immediate periorbital region and each has some characteristics of other disorders of the skin with which they can be confused. The diagnosis of sarcoids is, however, relatively simple in most cases. The lesions are not necessarily restricted to the dermis and epidermis, but commonly invade into the subcutis and the deeper muscular structures around the eye. Treatment with radiation is expensive and difficult to manage but resulted in the best outcome with almost 100% resolution in 66 cases. By contrast, surgical excision has serious potential complications that arise primarily as a result of the infiltrative nature of periorbital sarcoids. The value of BCG immunomodulation therapy for fibroblastic and nodular lesions described by previous workers is confirmed in this paper, with a good overall response (69%), provided that the material was injected intralesionally. Perilesional injection did not appear to carry a significant benefit. However, treatment of verrucose or occult lesions by this method gave generally poor results. The results of intralesional injection of cisplatin emulsions suggest that this may be an effective method of treatment in cases where other modalities are not feasible. In common with previous reports, failure to resolve the lesions frequently resulted in regrowth of the tumor, and in most cases this recurrence appeared to be more aggressive, with extensive local infiltration and faster growth. Conclusion The periorbital tissues are less tolerant of damage than those at many other sites and therefore certain treatment methods may be contraindicated. This suggests that it is important to select the best possible treatment at the first opportunity. It is a wise precaution to warn owners of horses with periorbital sarcoids of the dangers of leaving them and the particular risks associated with the currently available treatments. PMID- 11397302 TI - Systemic effects of topical and subconjunctival ophthalmic atropine in the horse. AB - OBJECTIVE: To identify any systemic effects of topical and subconjunctival administration of atropine sulfate in the horse. Animals studied Six mature grade horses were treated hourly in one eye with topical ophthalmic atropine drops for 24 h. Five horses were treated subconjunctivally in one eye with 3 mg of atropine sulfate. Procedures Pupillary light reflexes, pupil size, electrocardiographic parameters, girth measurements, intestinal motility, and clinical signs of abdominal pain were monitored. RESULTS: Alteration in auscultated gut motility and clinical signs of abdominal pain were the most sensitive indicators of the systemic manifestations of the topically applied atropine. Gut motility was absent in all horses for periods of 2-18 h in all four abdominal quadrants in horses given topically administered atropine. Signs of abdominal pain were observed in four of six horses that received topical atropine. In the subconjunctival test study, gut motility was absent in three horses for periods of 3-7 h. Uniocular subconjunctival injection of 3 mg atropine sulfate produced signs of abdominal pain in one of six horses. Conclusion The ophthalmic administration of atropine can affect gut motility and induce signs of colic in selected horses. PMID- 11397303 TI - Conjunctival malignant melanoma in a horse. AB - A case of malignant melanoma originating from the conjunctiva of a horse is reported. The tumor exhibited locally aggressive behavior as evidenced clinically by recurrence following two treatment episodes including surgical excision on each occasion and one application of cryotherapy. The orbit was subsequently exenterated and histologically malignant conjunctival melanoma was confirmed. Histopathologic features included variable pigmentation with amelanotic sites demonstrating marked cellular and nuclear pleomorphism with high numbers of mitotic figures. Cords of neoplastic cells invaded the sclera and cornea. Following exenteration, the horse exhibited no recurrence of the tumor for five years before being lost to follow-up. To our knowledge, this is the first report of primary malignant conjunctival melanoma in a horse. PMID- 11397304 TI - Corneal stromal sequestration and keratoconjunctivitis sicca in a horse. AB - A 19-year-old Shetland pony presented with unilateral ocular discomfort and abnormal ocular appearance. Keratoconjunctivitis sicca, ulcerative keratitis and brown discoloration of the corneal stroma were identified on ophthalmic examination. The etiology of keratoconjunctivitis sicca was not determined in this case. For practical and financial reasons, the owners requested enucleation of the affected eye. Histopathologic examination revealed extensive loss of corneal epithelium overlying a zone of hypereosinophilic, degenerate, and necrotic corneal stroma. This well-circumscribed region of corneal stromal sequestration was surrounded by stromal vascularization, and an intense inflammatory, predominantly polymorphonuclear, cellular infiltrate. The clinical and histopathologic features of this case were considered remarkably similar to those observed in feline corneal stromal sequestration. PMID- 11397305 TI - Orbital fibroma in a horse. AB - An 11-year-old American Quarterhorse gelding presented for moderate periorbital swelling and exophthalmia of the left eye. The menace response, and direct and consensual pupillary light reflexes were absent in the left eye. Conjunctival hyperemia, blepharedema, a mydriatic pupil, resistance to retropulsion, and an increased intraocular pressure were present. A soft-tissue mass could be palpated in the left retrobulbar space by pressing onto the orbit over the supraorbital fossa. Incomplete surgical resection of the mass was performed and histopathologic evaluation was consistent with a fibroma. Normal pupillary light reflexes and vision returned following surgery. The mass has not recurred 14 months after surgery. PMID- 11397306 TI - Practical usefulness of a therapeutic soft contact lens for a corneal ulcer in a racehorse. AB - Therapeutic soft contact lenses (TSCL) were applied to a thoroughbred racehorse with corneal ulcer refractory to topical medication. The insertion of the TSCL was conducted on day 36 using a nose twitch under sedation, auriculopalpebral nerve block, and topical anesthesia. The lens was retained, with the duration of wear lasting 114 days. Ocular pain improved immediately and fluorescein tests were negative 17 days after insertion. The horse was able to undergo training and ran in two races while wearing the TSCL. Thus a TSCL was found to be clinically useful for treating corneal ulcers in racehorses. PMID- 11397307 TI - Eosinophilic conjunctivitis, herpes virus and mast cell tumor of the third eyelid in a cat. AB - A 3-year-old Himalayan cat was diagnosed with concurrent eosinophilic conjunctivitis, herpes virus, and a conjunctival mast cell tumor. Eosinophilic conjunctivitis was verified via cytology from a conjunctival scraping, which revealed 50% eosinophils and 50% neutrophils. Herpes virus was verified via a positive polymerase chain reaction (PCR). Conjunctival scrapings for chlamydia immmunofluorescent antibody (IFA) and herpes IFA were negative. A mycoplasma was detected by a general mycoplasma PCR but the organism did not grow on the available mycoplasma media. The mass was excised and microscopic evaluation revealed a histiocytic mast cell tumor. The mast cell did not recur following local excision (at 1 year follow-up). The eosinophilic conjunctivitis was treated with both topical steroids and systemic megesterol acetate (Ovaban). When topical steroids were used, the herpes virus flared up and resulted in dendritic and geographic corneal ulceration. Therefore, the cat was treated with megesterol acetate and the eosinophilic conjunctivitis was well controlled. Treatment of eosinophilic conjunctivitis in the cat with megesterol acetate may be the treatement of choice due to the possibility of herpes virus. PMID- 11397308 TI - Lens-induced uveitis. AB - has been described in humans and many animal species. Traumatic rupture of the lens capsule may result in vision-threatening intraocular inflammation that is poorly responsive to medical management. Phthisis bulbi, persistent uveitis or glaucoma often occurs in these eyes. Surgical removal of the lens material is generally indicated shortly after the injury in an effort to preserve vision. Leaking of lens proteins through an intact lens capsule may result in a lympho plasmacytic anterior uveitis. This is most commonly associated with the presence of a hypermature cataract. The presence of lens-induced uveitis prior to cataract surgery significantly reduces the success rate of cataract surgery. Small amounts of circulating lens proteins maintain a normal T-cell tolerance for lens proteins. Lens-induced uveitis develops when a breakdown occurs of this normal T cell tolerance. Immune complexes play an important role in the tissue damage associated with the ensuing inflammation. Other factors associated with the tissue damage include hydroxyl radicals, nitroxide radicals, and hydrogen peroxide and arachidonic acid metabolites. Treatment consists of topical and systemic anti-inflammatory medications, mydriatic agents, and glaucoma medications when indicated. Experimental pharmacological agents include dual cyclooxygenase/lipoxygenase inhibitors, interleukin-1 blockers, antioxidants and hydroxyl radical scavengers. PMID- 11397309 TI - Retinal and optic nerve degeneration in cattle after accidental acrylamide intoxication. AB - A herd of cattle was accidentally exposed to monomeric acrylamide and N methylolacrylamide. Seven cattle underwent repeated examinations during 6-8 months after exposure. Abnormal pupillary light reflexes (PLR) were observed in one cow. Ophthalmoscopic examination showed progressive retinal degeneration and degenerative changes in the optic nerve heads in this cow. Light and electron microscopic examination revealed pathological changes in the retinae and optic nerves consistent with chronic stages of acrylamide toxicity. Ophthalmic examination and light microscopy did not reveal abnormalities in the retinae or optic nerves in either the rest of the cattle from the herd or in offspring of exposed cattle. PMID- 11397310 TI - Golden Retriever uveitis: 75 cases (1994-1999). AB - OBJECTIVE: To document the presenting ocular signs and the clinical course of Golden Retriever dogs with a progressive anterior uveitis, often associated with the histologic presence of iridociliary cysts. Animals studied Seventy-five Golden Retriever dogs (142 affected eyes) referred to a private practice referral ophthalmology clinic between 1994 and 1999. Procedures Complete ophthalmic evaluation with slit-lamp biomicroscopy, indirect ophthalmoscopy, applanation tonometry, and gonioscopy. Hematology, serum biochemical evaluations, and serologic titers for endemic infectious agents were also used in selected cases. RESULTS: The age range of affected dogs was 4.5-14.5 years, with a mean age of 8.6 +/- 2.1 years. The majority of the dogs (n = 66) were affected in both eyes at first presentation. The sex distribution included 4 intact males, 32 neutered males, and 39 spayed females. Hematology, serum biochemical evaluations, and serologic titers for endemic infectious agents failed to demonstrate any underlying disorder. The ophthalmic hallmark of this syndrome was the appearance of pigment on the anterior lens capsule, often in a radial orientation. This capsular pigment was seen both with and without associated uveal cysts. Although single to multiple iridociliary cysts were noted clinically in only 13.3% of the cases, cysts were common on histopathology of advanced glaucomatous, blind eyes. Fibrin was observed in the anterior chamber of 37% of the cases, and often was a precursor for glaucoma. Cataract formation (37%) and glaucoma (46%) were frequent sequelae to the uveitis. Posterior synechiae formation occurred in 50% of the cases. Histopathologic analysis of four enucleated eyes and the eviscerated specimens from 14 glaucomatous eyes demonstrated thin-walled iridociliary epithelial cysts in 3/4 and 12/14 cases, respectively. Microscopically, little to no uveal inflammatory infiltration was commonly noted. Conclusion The overall prognosis for this progressive uveitis in Golden Retriever dogs is guarded, with 46% of the eyes becoming blind due to glaucoma. Iridociliary cysts in Golden Retriever dogs may lead to the development of glaucoma in this breed. PMID- 11397311 TI - Towards a better canine intraocular lens. PMID- 11397312 TI - The effects of topical tropicamide and systemic medetomidine, followed by atipamezole reversal, on pupil size and intraocular pressure in normal dogs. AB - Twenty normal Golden Retrievers being screeened for eye, hip and elbow diseases were given tropicamide topically and medetomidine systemically. Medetomidine effects were later reversed with systemic atipamezole. Pupil size and intraocular pressure changes were determined. Pupil size increased significantly following tropicamide administration and continued to increase slightly but significantly after medetomidine injection. It was unclear whether the slight increase in pupil size following medetomidine administration was due to continued effect of tropicamide or due to the medetomidine itself. Atipamezole did not influence pupil size. Intraocular pressure (IOP) was not affected by these drugs. Ophthalmic screening examination for inherited disease following tropicamide administration is equally feasible prior to sedation with medetomidine and after reversal with atipamezole, but not during the period of sedation. PMID- 11397313 TI - Estimation of lacrimal level and testing methods on normal beagles. AB - Five methods of testing tears including the Schirmer tear test (STT-1), phenol red thread tear test (PRT) and modified Schirmer tear test (STT-2) were conducted on 44 eyes of 22 normal Beagles, and the tear film break up time (BUT) and lacrimal pH were tested in 32 eyes of 16 normal Beagles. The coefficient of variation of PRT demonstrated a low value (C.V. 12%) [corrected] compared to the value of STT-1 (C.V. 14%) [corrected]. The lacrimal pH showed a more constant value (C.V. 3%). Measurements of STT-2 (C.V. 48%) and BUT (C.V. 34%) varied significantly. The results show the clinical usefulness of the PRT. Mean values (mean +/- SD) of tests were: PRT, 29.3 +/- 3.45 mm/15 s; STT-1, 18.89 +/- 2.62 mm/60 s; STT-2, 9.52 +/- 4.55 mm/60 s; pH, 7.29 +/- 0.22 and BUT, 21.53 +/- 7.42 s. PMID- 11397314 TI - The effect of third eyelid gland removal on the ocular surface of dogs. AB - To evaluate the effect of third eyelid removal on the ocular surface of dogs, we operated on five young Beagle dogs and observed changes to tear function using the following tests: phenol red thread test (PRT), Schirmer tear test (STT-1), modified Schirmer tear test (STT-2), pH and tear break-up time (BUT). There was a significant decrease (37%) in STT-2 within 2 weeks after the excision and this declined further to 60% at 1 year. The pH value increased after excision. Presurgical pH was 7.17 +/- 0.20 (mean +/- SD), which increased to 7.55 +/- 0.24 in the 14-60 days following removal, and further increased to 7.77 +/- 0.65 at 1 year. The PRT and STT-1 decreased by 26% within 3-7 months compared to pre excision values, but by 1 year the values recovered to near normal. The BUT pre excision value was 24.0 +/- 8.1 s, which shortened to 13.5 +/- 4.5 s after 5 months and continued to decrease further during the study. There were no overt visual signs of KCS during the observational period. However, microinjury of the keratoconjunctival epithelium was observed for all operated eyes when vital staining was used at 1 year post surgery. Surgical excision of the third eyelid in Beagle dogs influenced tear quality level and affected the stability of the tear layer, and at 1 year there was evidence of microinjury to the keratoconjunctival epithelium. PMID- 11397315 TI - Retrobulbar tumors in dogs and cats: 25 cases. AB - Twenty-five cases of retrobulbar tumors are presented and discussed. Affected animals were dogs and cats (average 10.7 years). No breed or sex predisposition was noted. The most common clinical signs were exophthalmos (84%), conjunctival hyperemia (40%), protrusion of the nictitating membrane (28%), exposure keratitis (20%) and fundus abnormalities (20%). Diagnostic tools included fine needle aspiration, radiography, ultrasonography, computed tomography and histology. Surgical treatment by orbitotomy or exenteration was combined with chemotherapy and radiotherapy in some cases. The prognosis was poor with low survival times: 1 month in cats, and 10 months in dogs, with a high rate of euthanasia (35%) at the time of diagnosis. PMID- 11397316 TI - Persistent fetal intraocular vasculature in the European ferret (Mustela putorius): clinical and histological aspects. AB - OBJECTIVE: To describe the clinical and histological appearance of persistent fetal intraocular vasculature in a colony of ferrets. Design Prospective study. ANIMALS STUDIED: Eighty-six European ferrets (Mustela putorius). Procedure Both eyes of 76 genetically related progeny and 10 breeding, adult, colony-raised ferrets were studied using a slit lamp biomicroscope and an indirect ophthalmoscope. Ferret progeny were examined after eyelid opening at 5-6 weeks of age, and at 12 months of age. After euthanasia, globes were enucleated and examined histologically. RESULTS: Persistent fetal intraocular vasculature was evident in 21 progeny ferrets at 5-6 weeks of age and in three mature progenitor ferrets. Clinical appearance of diminutive vasculature was characterized by focal remnants of the posterior tunica vasculosa lentis, muscae volitantes, and an occluded hyaloid artery extending from the optic papilla and terminating in the anterior vitreous body. Extensive persistent vasculature was characterized by a perfused hyaloid artery, vasa hyaloidea propria and posterior tunica vasculosa lentis, posterior cortical and capsular cataract, and proliferation of fibrovascular tissue along the posterior lens capsule. Fetal vasculature persisted in 7 of 21 progeny ferrets at one year of age and in three progenitor ferrets. Results of histologic examination showed persistence of the hyaloid vasculature, proliferation of retrolental fibrovascular tissue with osseous metaplasia, posterior capsular and cortical cataract, and occasional retinal detachment. CONCLUSIONS: Persistent fetal intraocular vasculature in ferrets appears similar clinically and histologically to persistent fetal intraocular vasculature reported in humans and dogs. The ferret may be a suitable animal model for vasculogenic mechanisms of persistent fetal intraocular vasculature and for evaluating vasoinhibitory growth factors and angiostatic test compounds. PMID- 11397317 TI - Ocular complications of persistent hyperplastic primary vitreous in three dogs. AB - Persistent hyperplastic primary vitreous (PHPV) syndrome associated with either severe ocular complications or multiple ocular lesions was diagnosed in three young dogs, a Samoyed, a Spanish Pachon, and a mixed breed dog. Due to opacification of the anterior ocular structures, B-mode and color-flow Doppler ultrasonography were performed to aid diagnosis. The Samoyed presented with unilateral hyphema; the Spanish Pachon presented with unilateral secondary glaucoma associated with uveitis and hyphema OD and leucocoria OU; and the mixed breed presented with bilateral leucocoria. B-mode ultrasonography of the Samoyed revealed a subcapsular cataract and a hyperechoic tubular structure attached from the optic disk to the posterior lens capsule. In the Spanish Pachon B-mode ultrasonography of the right eye indicated microphakia, cataract formation, and a retrolental mass with a thin hyperechoic strand stretching from the optic disk to the posterior lens; and for the right eye cataract formation, PHPV, retinal detachment, and vitreous hemorrhage. In the mixed breed dog, B-mode ultrasonography of both eyes indicated microphthalmia, retrolental mass, and hyperechoic lenses. By color-flow Doppler imaging, blood flow was present in the retrolental mass of the right eye suggesting a persistent hyaloid artery. PMID- 11397318 TI - Ocular fundus images with confocal scanning laser ophthalmoscopy in the dog, monkey and minipig. AB - Confocal scanning laser ophthalmoscopy (CSLO) is a new technique that enables ocular fundus image recording and retinal dynamic angiography to be performed. The ocular fundus image is acquired sequentially, point by point, and is reconstructed on a video monitor at the rate of 25 images per second. The feasibility of performing both ocular fundus image recordings and retinal angiography image recordings were tested on two dogs, two monkeys and two minipigs using a 40 degrees field I + Tech CSLO. Fundus area of each dog, monkey and minipig were examined without any additional optical devices. The ocular fundus and angiography images were recorded, stabilized and analyzed under the same conditions. For each species, all images were easily recorded without any additional optical device in a lighted room and the morphology of the retinal images generated was similar to those obtained with a camera or angiography of higher resolution. Capillary phase or venous times are presented. Image recording at 25 frames/second enabled more retinal dynamics to be demonstrated than with use of regular angiography. This technique is noninvasive and easy to perform if the eye is fixed and eyelids maintained open. It also allows exploration of retinal microvascularization and could be utilized for clinical, pharmacologic and toxicologic investigations as well. PMID- 11397319 TI - Schwalbe line's cell in the normal and glaucomatous dog. AB - OBJECTIVE: The canine iridocorneal angle contains an operculum, which is similar to that in nonhuman primates and consists of a peripheral extension of the inner cornea that overlies the anterior-most portion of the corneoscleral trabecular meshwork. This region contains cells, the Schwalbe line's (SL) cells, that have been found to have secretory and epithelial characteristics. This region of the iridocorneal angle represents the nonfiltering portion and becomes altered early during spontaneous glaucoma in the Beagle. The present study describes the SL cell for the first time in the dog and changes associated with canine primary open angle glaucoma. PROCEDURES: The iridocorneal angles from 18 Beagles with inherited glaucoma (3 months-8 years old) and 17 normal, age-matched Beagles were placed in 10% buffered formalin for light microscopic evaluation, or 2.5% glutaraldehyde for ultrastructural evaluation. Using at least three fields from each region of the iridocorneal angle (opercular, corneoscleral, and uveal) at x 1000 magnification, trabecular cell nuclei were counted. RESULTS: The operculum in the canine iridocorneal angle consisted of the peripheral extension of the corneal endothelium and underlying anterior-most corneoscleral meshwork, having no direct contact with the angular aqueous plexus. The SL cells associated with operculum-retained epithelial morphology (polyhedral in shape with rER, Golgi apparatus, and secretory vesicles) in both normal, pre-and early glaucomatous dogs. In animals with moderate and advanced stages the SL cells often became less epithelial and secretory in appearance. The number of SL cells in normal dogs declined by approximately one-third by the end of their first year with gradual loss thereafter. In the glaucoma group the decline was more substantial and continuous through the first three years. CONCLUSIONS: The SL cell is morphologically a distinct cell type within the canine iridocorneal angle that is specifically associated with the nonfiltering portion of the corneoscleral trabecular meshwork. Changes in the SL cells of the glaucomatous dog occurred with regard to age and progression of the disease. PMID- 11397320 TI - Microvasculature of the rat eye: scanning electron microscopy of vascular corrosion casts. AB - Methylmethacrylate castings of the eye microvasculature were prepared from 10 Spargue-Dawley rats and studied by electron microscopy. The choroidal arterioles are larger in diameter than retinal arterioles, and have a shorter course to choroidal capillaries than retinal arterioles to retinal capillaries. Retinal capillaries are extremely thin in diameter and form a sparse retinal capillary network. The choriocapillaris is large and sinusoid-like, forming a compactly arranged network in the choroid. These differences in the microvasculature between the choroid and retina help explain the differences in ocular hemodynamics; that is, the blood flow in the choroid is faster than that in the retina. Capillaries of the iris show a zigzag configuration, which may be an accommodation for dilation and constriction of the pupil. Capillaries of the ciliary body are of large diameter forming leaf-like configurations, presumably to contribute to the secretion of the aqueous humor. Capillaries of the conjunctiva exhibit a somewhat coiled configuration, the arrangement of which reduce tension of the conjunctiva vessels caused by eyeball movement. Intra arterial cushions, which control blood flow at the branching sites, are found in both choroidal or retinal arterioles. PMID- 11397321 TI - Changes in intraocular pressure associated with topical dorzolamide and oral methazolamide in glaucomatous dogs. AB - OBJECTIVE: To compare the reduction in intraocular pressure (IOP) by topical 2% dorzolamide to oral methazolamide (5 mg/kg) in dogs, and determine if the combination of both drugs would reduce IOP more than either drug administered alone. ANIMALS STUDIED: Thirteen glaucomatous beagles. PROCEDURES: Measurements, including applanation tonometry, pupil size and heart rate, were obtained at 8 am, 12 noon, and 5 pm on days 1, 3 and 5. The 5-day drug studies included placebo (0.5% methylcellulose); 2% dorzolamide administered in one eye twice daily (8 am and 5 pm), and repeated again in one eye three times (8 am, 12 noon and 5 pm) daily; methazolamide (5 mg/kg per os administered at 8 am and 5 pm); 2% dorzolamide instilled twice daily (5 days) combined with oral methazolamide on the last 3 days, and methazolamide (5 days) combined with 2% dorzolamide on the last 3 days and instilled twice daily. Statistical comparisons between drug groups included control (nondrug) eye and treated (placebo/drug) eyes for days 1, day 3 and 5. RESULTS: Topical 2% dorzolamide, administered twice and three times daily, significantly decreased IOP (mean +/- SEM) in glaucomatous dogs on the first day (twice daily 7.6 +/- 2.4 mmHg, and three times daily 16.4 +/- 3.6 mmHg) that was even greater by day 5 (twice daily 10.4 +/- 2.0 mmHg, and three times daily 13.9 +/- 2.7). Oral methazolamide also significantly lowered IOP in both eyes. Oral methazolamide (administered from day 1 through to day 5) combined with 2% topical dorzolamide (instilled in the drug eye for day 3 through to day 5) also significantly lowered IOP of both eyes for all days, and for day 5 the mean +/- SEM IOP was decreased by 7.9 +/- 1.7 mmHg (methazolamide plus dorzolamide) and 7.5 +/- 2.6 mmHg (methazolamide only). Topical dorzolamide (instilled in the drug eye for day 1 through to day 5) combined with oral methazolamide (administered from day 3 through to day 5) significantly lowered IOP in the drug eye on day 1 (5 pm: 9.6 +/- 1.9 mmHg), for day 3 (11 am and 5 pm) and for all of day 5 for both eyes (5 pm: control eye 9.5 +/- 1.8 mmHg; drug eye 9.2 +/- 1.9 mmHg). Topical dorzolamide (2%) instilled three times daily produces similar IOP declines compared to the combination of oral methazolamide and 2% dorzolamide administered twice daily. CONCLUSIONS: Dorzolamide (2%) instilled twice or three times daily causes significant decreases in IOP in glaucomatous dogs. Twice daily instillations caused progressive declines in IOP from day 1 to day 5. Dorzolamide (2%) combined with oral methazolamide (5 mg/kg per os twice daily) produces similar but not additional declines in IOP. PMID- 11397322 TI - Feline eosinophilic conjunctivitis. AB - OBJECTIVE: To review 12 cases of histologically confirmed feline eosinophilic conjunctivitis, their clinical, cytologic, histologic and electronmicroscopic findings, results on PCR for FeHV-1, treatment and outcome. ANIMALS STUDIED: Twelve naturally occurring cases presented during a period of 26 months. PROCEDURES: Thorough ophthalmologic examination, conjunctival scrapings performed with the cytobrush method; histologic samples from the palpebral conjunctiva; PCR for FeHV-1 on Schirmer Tear Test (STT) strips; saliva and nasal swabs, and retrospective evaluation of all results. RESULTS: The breed most commonly affected was the Domestic Shorthair (n = 8), followed by Persians (n = 2), Somali (n = 1) and Siamese (n = 1). Age at presentation was 1-15 years with a mean age of 7.2 years. Nine cats were castrated males; three cats were females: two of them were spayed. Unilateral (n = 7) or bilateral (n = 5) involvement with depigmentation and erosions of lid margin, blepharospasm, swelling and redness of conjunctiva and third eyelid were the most common clinical findings. Frequency of eosinophils in cytologic samples was more than 10% in every patient. PCR for FeHV 1 on STT was negative in all cases. Histologically, eosinophils, lymphocytes, plasma cells, mast cells and macrophages were involved. On electronmicroscopy, viral particles were not detected. Ten cases needed long-term anti-inflammatory treatment. CONCLUSIONS: The 12 reviewed cases suggest that feline eosinophilic conjunctivitis is a chronic inflammatory uni- or bilateral disease of the adult cat. Typically the lid margin was also involved, and was thickened, depigmented and erosive. Cytological examination of conjunctival scrapings was a valuable tool for detecting eosinophilic conjunctivitis. The cytological findings correlated well with the histopathological findings in our patients. Topical or systemic anti-inflammatory drugs resolved the clinical symptoms in our cases within a short period of time. Neither electronmicroscopy nor PCR were able to detect involvement of FHV1 in the represented cases. The etiopathogenic role of FeHV-1 remains undetermined. PMID- 11397323 TI - Suspected latent vertebral metastasis of uveal melanoma in a dog: a case report. AB - A six-year-old intact male Pomeranian was examined because of right eye discomfort. An iris neoplasm was suspected and the eye was enucleated. A uveal melanoma with malignant features was diagnosed. The dog recovered uneventfully from surgery. A general physical examination was performed at 3-month intervals afterwards without any detectable problem, but 18 months after the first presentation the dog suffered a rapid, progressive paraplegia. Radiographic examination and myelography revealed a spinal cord compression at the level of the 8th thoracic (T8) vertebral body. Surgical exploration of the area revealed a potential vertebral neoplasm: histopathology confirmed a melanoma which was suspected to have resulted from metastasis from the previously diagnosed uveal melanoma. PMID- 11397324 TI - Platelet count and interleukin 6 gene polymorphism in healthy subjects. AB - BACKGROUND: Interleukin 6 (IL-6) is thought to play important roles in the development of reactive thrombocytosis caused by inflammation by its stimulatory effect on megakaryocytopoiesis. A G/C polymorphism of the IL-6 gene at position 174 has been found to be associated to different transcription rates. Specifically, subjects with the CC genotype showed lower plasma IL-6 levels compared with GC or GG subjects. Given this difference in transcription rates of IL-6 we speculated on different platelet count according to this IL-6 polymorphism. METHODS: The G/C polymorphism of the IL-6 gene at position -174, serum IL-6 concentration and platelet count were prospectively analyzed in 59 (25 women) consecutive healthy subjects. RESULTS: Subjects who were homozygotes for the C allele at position -174 of the IL-6 gene (Sfa NI genotype) showed significantly lower platelet count than carriers of the G allele, despite similar age, sex, body mass index and proportion of smokers (205400 +/- 44088 vs 239818 +/- 60194, p = 0.047). This was in parallel to differences in peripheral white blood cell count (5807 +/- 1671 vs 6867 +/- 1192 x 10(9)/ml, p = 0.01). CONCLUSION: This is the first description, to our knowledge, of a genetical influence on basal platelet counts, which appears to be partially dependent on a polymorphism of the IL-6 gene, even in the absence of inflammation. PMID- 11397325 TI - The interactivity between the CFTR gene and cystic fibrosis would be limited to the initial phase of the disease. AB - PURPOSE: Cystic fibrosis (CF) is a lethal genetic disorder affecting secretory epithelia, caused by mutations on the CFTR gene. In this paper we study the interactivity between the CFTR gene and CF disease over the time course of CF. METHOD: Cross-sectional analysis of CF patient population data from Latin America, Canada, and The Netherlands, under the assumption that they represent stationary populations, was used to determined and correlates hazard rates, average cores and CF progression rates. RESULTS: Results suggests the existence of two phases throughout the course of CF. CONCLUSION: While the initial phase was related to the CFTR genotype, the kinetics of the second phase seems to be common to all groups considered. The hypothesis that the interactivity between the CFTR gene and CF disease would be limited in time is presented, suggesting that mutant CFTR would trigger a disease that evolves to become independent from the CFTR gene itself. PMID- 11397326 TI - Characteristics of two cases with dup(15)(q11.2-q12): one of maternal and one of paternal origin. AB - PURPOSE: The phenotype correlations for interstitial duplications that include the Prader-Willi/Angelman syndrome critical region are not well established. We describe two such duplication cases, one of which was of maternal origin and the other was paternal. METHODS: High resolution G-banding, fluorescence in situ hybridization (FISH) for SNRP-N and D15S10 were used for cytogenetic analysis. Southern blot analyses based on parent of origin specific DNA methylation at D15S63 (PW71) locus were utilized for detection of methylated and unmethylated fragments. RESULTS: The duplication was established by the FISH analysis. The molecular pattern suggested a maternal origin of the duplication in patient 1 and a paternal origin in patient 2. Patient 1 (2 years old) had developmental and speech delays with pervasive developmental disorder or mild autism, strabismus, and normal growth parameters with seizures. Patient 2 (16 years old) had global developmental delay, verbal IQ of 94, depression, obesity, food-seeking behavior, and significant behavioral problems that included self-injurious tendencies. Neither patient had significant dysmorphic features or abnormalities of internal organs. CONCLUSION: The two cases suggest that some patients with 15q11.2q12 duplication may have significant anomalies, and there appear to be phenotypic differences between maternal and paternal transmission of the duplication. PMID- 11397327 TI - Molecular determination of X inactivation pattern correlates with phenotype in women with a structurally abnormal X chromosome. AB - PURPOSE: To correlate the X inactivation pattern, as determined by one or more molecular assays, with phenotype in individuals with structurally abnormal X chromosomes. METHODS: We utilized methylation analysis of androgen receptor (AR) and Fragile X (FMR1) genes and expression studies of an XIST polymorphism to assess X inactivation patterns of 28 females with structurally abnormal X chromosomes. Individuals were placed in one of three categories: (1) completely nonrandom inactivation of one X chromosome, (2) preferential or skewed inactivation of one X chromosome, or (3) random inactivation of either X chromosome. RESULTS: In 19 of 21 cases with complete (>97%) skewing of X inactivation, the phenotype was either normal, consistent with a single gene disorder, or consistent with classical Turner syndrome; two cases with completely nonrandom X inactivation had unexplained mental retardation phenotypes. In contrast, six of seven cases that did not exhibit completely nonrandom X inactivation were phenotypically abnormal. Carriers of two balanced translocations, two duplicated Xs, one deleted X, and one 45,X/46,X,r(X) presented with mental retardation and/or multiple congenital anomalies. CONCLUSION: In patients with random or skewed X inactivation, the abnormal phenotype was hypothesized to be due to functional nullisomy or disomy of X linked genes. Based on these results, we propose that X inactivation studies should be performed on all women with structurally abnormal X chromosomes. This should aid in the understanding of abnormal phenotypes in liveborn individuals with abnormal X chromosomes and may help to predict phenotypes for prenatally detected cases in the future. PMID- 11397328 TI - Outcomes analysis of verbal dyspraxia in classic galactosemia. AB - PURPOSE: This study evaluates a genotype/phenotype relationship between developmental verbal dyspraxia (DVD) and the common, missense mutation of the galactose-1-phosphate uridyltransferase gene, Q188R, in patients with classic galactosemia (G/G). METHODS: As part of this study, we devised a questionnaire for "speech problems" to be completed by the patient?'s clinician. To validate the questionnaire and determine its accuracy in detecting DVD, we analyzed questionnaire responses for 21 patients by testing them independently and directly for DVD through a speech pathologist blinded to the patients' genotype. RESULTS: We found that the questionnaire had a sensitivity of 0.56 and a specificity of 0.75. We then calculated the prevalence of DVD for a larger set of 113 patients with G/G galactosemia whose biochemical phenotype, molecular genotypes, and clinical status were known. The prevalence of "speech problems" from raw data were 50 of 113 (44.2%). After adjusting for misclassification, 43 (38.1%) were classified as cases of DVD. Using multivariate, logistic, regression analyses we found a significant interaction between genotype and mean red blood cell (RBC) galactose-1-phosphate (Gal-1-P). When corrected, using mean RBC Gal-1 P < h 3.28 mg%, the Q188R/Q188R genotype was the best predictor of DVD. There was a significant risk (odds ratio = 9.6, p = 0.0504) of having DVD associated with homozygosity for Q188R compared with other genotypes. CONCLUSIONS: We conclude that homozygosity for Q188R mutations in the GALT gene is a significant risk factor for DVD. However, poor metabolic control obviates this relationship as indicated by RBC Gal-1-P greater than 3.28 mg%. PMID- 11397329 TI - Gay men's casual sex encounters: discussing HIV and using condoms. AB - Some gay men disclose their HIV serostatus to enable them to decide to discard condoms in at least some casual encounters. In this paper we consider under what circumstances this occurs. We draw on data from the Sydney Men and Sexual Health cohort study, including both structured and semi-structured interviews. We find that casual sexual encounters among gay men are complex events, involving a broad range of types of partner and circumstance. A significant minority of men disclose their HIV serostatus; 36.3% of those who discussed their serostatus with casual partners engaged in unprotected anal intercourse (UAI) with those partners, while only 9.9% of those who did not discuss their serostatus engaged in UAI with those partners. We also find that disclosure of HIV serostatus, discussion and use of condoms during casual encounters are affected by the particular circumstances of those encounters, including the degree of familiarity between casual partners. PMID- 11397330 TI - Consistent condom use in the heterosexual relationships of young adults who live in a high-HIV-risk neighbourhood and do not use "hard drugs". AB - This study was set up to determine the predictors of condom use in the heterosexual non-commercial sexual relationships of young adults who neither inject drugs nor use cocaine, heroin or crack, in a neighbourhood with widespread drug-use-connected HIV. The analytic sample is 279 young adults, aged 18-24, who have never injected drugs and who have not used heroin, cocaine or crack in the last year. They were recruited in the Bushwick neighbourhood of New York City, July 1997 to September 1999. A face-to-face interview included items about their sociodemographic background, substance use and sexual networks. Sexual relationship and self-reported consistent (100%) condom use over the prior year with the partner in a given relationship was examined. Subjects had 337 heterosexual non-commercial relationships. Consistent condom use was reported in 32% of these relationships. In multiple logistic regression, consistent condom use was more likely in relationships that are not 'very close' (odds ratio = 3.92; 95% confidence interval = 2.08, 7.52); in the relationships of subjects whose peer norms support condom use (OR = 1.94; 95% CI = 1.43, 2.69), who are not problem drinkers (OR = 8.70; 95% CI = 2.22, 58.8), and (perhaps as a result of measurement issues) who are men (OR = 1.95; 95% CI = 1.04, 3.68). In conclusion, consistent condom use remains uncommon among youth in this high-risk neighbourhood. It is thus important to keep HIV from entering the sexual networks of youth in communities like this through programmes aimed at drug injectors and their sexual partners. Programmes to increase condom use among young adults should focus on strengthening norms that promote safer sex to protect oneself and others. In addition, assistance should be provided to youth who are problem drinkers. PMID- 11397331 TI - Understanding constraints to adolescent condom procurement: the case of urban Botswana. AB - Even when condoms are widely available and affordable, adolescents may be reluctant to obtain them. Hence, programme managers need to understand what determines youths' perceptions of access to condoms. This paper analyzes focus group and survey data on condom access conducted among male and female adolescents in urban Botswana. Although a majority of sexually experienced adolescents have purchased condoms from retail outlets, only about 50% have obtained condoms from health facilities, even though the latter distribute free condoms. This study shows that many adolescents perceive that access to condoms is more difficult from public sector outlets than for private sector outlets, because the public sector providers tend to question the adolescents' behaviour while the latter do not. To bypass this problem, males tend to obtain condoms from friends. However, females are reluctant to ask their friends for condoms because they fear that their friends may gossip about them. The results of this study indicate that adolescents' access to condoms can be improved by interventions that improve the quality of interaction between public sector providers and adolescents, destigmatize condom use, expand private sector condom distribution, and that use peer sales agents and educators. PMID- 11397332 TI - 'Don't know' and 'didn't think of it': condom use at first intercourse by Latino adolescents. AB - We examined the specific reasons Latino adolescents did or did not use condoms at first intercourse and their specific reasons for their perceived risk for contracting HIV. Latino adolescents (n = 618), ages 11-19, completed a face-to face interview that included information on demographics (sex, age, family-status (lives with both natural parents versus other family structure) and country of birth) and sexual behaviour. The respondents cited 'don't know' (25.9%), 'not available' (25.9%) and 'didn't think of it' (23.5%) as the most frequent reasons for not using condoms at first intercourse. Anyone can get it (41.8%), unknown knowledge of partner's serostatus (24.7%) and unprotected sex (23.8%) were the top three reasons for perceiving oneself as being at risk for contracting HIV. Sexually active adolescents were more likely to perceive themselves at risk for contracting HIV than adolescents that had not had sex. Males were significantly more likely to report using condoms for protection at first intercourse than females. The remaining demographic factors were not significantly related to use of condoms at first intercourse nor to perceived risk for contracting HIV. PMID- 11397333 TI - The additional value of anticipated regret and psychopathology in explaining intended condom use among drug users. AB - Abstract In this study we examine determinants of intended condom use with casual and steady sex partners among drug users. Our aim was to find out whether anticipated regret and psychopathology represent additional factors in explaining condom use behaviour, on top of the constructs defined in traditional psychosocial models of behaviour. A questionnaire measuring intentions, attitude, beliefs, personal norm, subjective norm, descriptive norm, self-efficacy and anticipated regret toward condom use with steady and casual sex partners, and the Symptom Checklist '90 (to measure dimensions of psychopathology) were handed out to 150 Dutch drug users. The results showed that intended condom use with steady sex partners was mainly predicted by self-efficacy, personal norm and anticipated regret (total R2 = 0.41). Lower feelings of hostility and a more positive personal norm, subjective norm and attitude significantly predicted intended condom use with casual sex partners (total R2 = 0.24). The results also showed that intended condom use with steady sex partners seems to result from anticipating possible barriers, whereas condom use with casual sex partners is largely a matter of having safe sex norms. Practical implications of this study for Aids-preventive campaigns are also discussed. PMID- 11397334 TI - Correlates of quality of life in symptomatic HIV patients living in Hong Kong. AB - This cross-sectional study assessed quality of life (QOL), coping styles, mood and uncertainty in illness in a non-random sample of 46 (out of 91 eligible) symptomatic HIV patients living in Hong Kong. QOL was moderate and the main concerns were related to the environmental aspects of QOL, spirituality and social relationships. Considerable mood disturbance was demonstrated in the sample, especially with regards to depression, fatigue and tension/anxiety. High levels of uncertainty in illness were also reported. A median split of the uncertainty score demonstrated that high uncertainty was related to lower levels of overall QOL (p = 0.04), higher psychological dysfunction (p = 0.05), worse adjustment with the environment (p < 0.001) and higher mood disturbance (p = 0.008). The sample predominantly used internal coping, which also correlated well with higher QOL scores. Through regression analysis it was shown that QOL could be predicted with the combined effects of uncertainty in illness and fatigue (adjusted R2 = 0.51, p < 0.001). Findings indicate that efforts should be directed towards improving QOL issues in the Chinese HIV patients and interventions could be introduced to alleviate those factors that were found to affect QOL. It is suggested that such interventions may include group or individual psychological therapies, management of fatigue and teaching patients more effective coping techniques. PMID- 11397335 TI - The role of family and friend social support in reducing emotional distress among HIV-positive women. AB - The purpose of this study was to examine HIV-positive women regarding their perceptions of family and friend social support and mental health outcomes. Regression models were constructed for five mental health outcomes. Results indicated that while each outcome has slightly different significant predictors, perceived family support was predictive of the reduced loneliness over the past few days and past year, stress, and presence of depressive symptoms. Implications for researchers and therapists are discussed. PMID- 11397336 TI - HIV infection and depressive symptoms: an investigation of African American single mothers. AB - Risk for depressive symptoms among HIV-infected African American single mothers (n = 96), relative to demographically matched non-infected single mothers (n = 120), was examined, using both self-report and clinician-rated scales of depression. Assessment of depressive symptoms occurred at two points separated by one year. Findings revealed that HIV-infected mothers were at greater risk for depressive symptoms at both assessments, regardless of method of assessment. Moreover, HIV-infected mothers remained at greater risk when analyses were limited to cognitive and affective symptoms of depression, decreasing the likelihood that the difference between the two groups was due to greater endorsement of somatic symptoms of depression by the HIV-infected group. PMID- 11397337 TI - Coping with HIV/AIDS in Durban's commercial sex industry. AB - This paper describes coping mechanisms used by commercial sex workers (CSWs) and their partners in confronting the threat of HIV. Data are part of a study exploring sexuality and HIV-related issues among members of the Durban commercial sex industry. Participants were 100 female CSWs, 25 male trucker driver clients and ten male personal partners. Data were collected using semi-structured questionnaires, focus group discussions and in-depth interviews. Analysis revealed high HIV-awareness and high prevalence of risky sexual behaviour. While they were acutely aware of the sex industry's potential role in HIV spread, study participants chose to remain sexually involved and engage in high risk sexual practices with both professional and personal partners. Men and women adopted several strategies to cope with the possibility of HIV infection: (1) denial of risk, (2) fatalism, (3) economic rationalization, (4) partner categorization through selective condom use, (5) purposeful ignorance of HIV status, and (6) abnegation of responsibility for practising safe sex. Among the most significant findings is the difference in study participants' handling of HIV risk and employing coping mechanisms in personal versus professional sexual situations. The implications of these coping strategies for HIV education, message development and intervention in the commercial sex industry and in general are discussed. PMID- 11397338 TI - Prevalence of HIV infection and risk behaviour among street prostitutes in Rome, 1997-1998. AB - A cross-sectional survey was carried out between April 1997 and February 1998 among street prostitutes in Rome. The study population (n = 142) consisted of 102 women and 40 transsexuals: 20% from Western Europe, 38% Eastern Europe, 23% Latin America and 17% Africa. Two-thirds of the population had more than 20 clients during the last week and most respondents (95%) reported always using condoms with clients. Eight per cent of the women and 2% of the transsexuals report a history of injecting drug use. Only 38% of the women with a stable partner reported use of contraceptives and 33% of them had undergone a voluntary abortion during the last year. Only 38% of the women had been checked for STDs during the last year, compared with 80% of the transsexuals. The HIV-prevalence was 6% among the women and 20% among the transsexuals. Four out of the six positive women and one of the positive transsexuals had a history of injecting drug use. Five out of the six HIV-positive women were Italian. Transsexual prostitutes seem to pay more attention to their medical wellbeing compared with females who rarely go for medical check-ups and only a minority uses efficient contraceptive methods. PMID- 11397339 TI - Truck drivers, middlemen and commercial sex workers: AIDS and the mediation of sex in south west Uganda. AB - Although long distance truck drivers have been implicated in the spread of HIV in Africa, there is a paucity of studies of their sexual cultures. This paper reports on a study of the sexual culture of drivers, mediators and commercial sex workers (CSWs) in a roadside truck stop on the Trans-Africa highway in south west Uganda. Sixty-nine truck drivers, six middlemen and 12 CSWs were interviewed using semi-structured questionnaires. Interviewing truck drivers also entailed participating in the town's nightlife and spending much time in the bars. Truck drivers stop briefly at the truck stop for various reasons: to eat, sleep, have sex and sell goods they are carrying. Middlemen mediate the latter two activities. Middlemen buy goods from the drivers and introduce them to 'suitable' women with whom they can have casual sex. Most drivers have sex when they spend the night at the truck stop, and most make use of the services of the middlemen. The most important reasons why drivers use middlemen are that the latter speak the local languages and, in particular, know the trustworthy and 'safe' (HIV negative) women. The CSWs use middlemen mainly because they are a guarantee that the driver will pay and they usually ensure that drivers pay well. The mediation system is becoming increasingly professionalized. Most drivers claimed to use condoms during casual sex, and this was confirmed by the CSWs. General use of condoms is encouraging, particularly given the context of a culture generally opposed to condoms. The idea that middlemen can recognize 'safe' women is worrying. However, given their key position, middlemen could form the hub of an opinion leader type intervention focused on drivers and the professional group of sex workers described here, providing condoms, advising about the importance of condom use in all casual sexual encounters, giving information about HIV and STDs, and possibly referring drivers and women to appropriate sources of HIV counselling and testing and STD treatment. PMID- 11397340 TI - Knowledge, risk perceptions and condom usage in male sex workers from three Australian cities. AB - The study identifies factors associated with knowledge and perception of risk of HIV/AIDS, as well as attitudes to and usage of condoms by a sample of male sex workers (MSW). One hundred and eighty-five male sex workers completed a self reported questionnaire, including knowledge about HIV transmission, attitudes to condom use and perceptions and personal susceptibility to HIV and sexually transmitted infection (STI) risk, and a two-week diary recording use of condom during commercial sex encounters. The findings reveal that condom use was found in 77.7% of the encounters with clients and the majority of the respondents perceived themselves to be at no risk for HIV because of sex work. Independent sex workers from Melbourne and workers who owned their place of residence used condoms in a significant lower proportion. Generally speaking, knowledge about the risks associated with AIDS was high, with respondents showing lower knowledge about the risks associated with unprotected receptive or active oral sex. Participants held a positive attitude to condom use; most considered the provisions of condoms to be their responsibility rather than clients; and they were more worried about contracting an STI than HIV. Those who scored higher on the knowledge scale had more positive attitudes to condom use and those who had a more positive attitude to condom use recorded a perceived lower risk of contracting STI but not HIV. The study discusses the relevance of these findings for public health risk reduction and sexual health education campaigns. PMID- 11397342 TI - UNAIDS/WHO global AIDS statistics. PMID- 11397343 TI - Recent trends in fetal and infant outcomes following post-term pregnancies. AB - All births and infant deaths in 1985 87 and 1992 94 in Canada, except in Ontario and Newfoundland, were analyzed to assess the potential impact of the recent increased use of elective labour induction for post-term pregnancies. Probabilistic linkage was carried out of infant death records (Canadian Mortality Database) and respective birth registrations (Canadian Birth Database) for the periods 1985 87 and 1992 94. The combined fetal and infant mortality declined by 20 30% between 1985 87 and 1992 94 at each gestational week beginning at 37 weeks, with no increased reduction among post-term pregnancies. Asphyxia-related fetal and infant deaths, the most likely cause of death being preventable by labour induction for post-term pregnancies, did not decrease among post-term pregnancies. In contrary, a substantial decrease of asphyxia-related deaths was observed at 37 and 38 weeks over the same periods of time. Because fetal and infant deaths are rare events and because the number of pregnancies passing 42 weeks of gestation decreased dramatically during 1992 94, statistically unstable results may be inevitable in the comparison of mortality in this group of pregnancies. PMID- 11397344 TI - The duration of major depressive episodes in the Canadian general population. AB - The National Population Health Survey (NPHS) has provided a wealth of new data concerning major depression in the Canadian general population. The NPHS included a brief predictor of major depression, and also two questions (only one of which was asked of each subject) concerned with the duration of episodes in the preceding year. A striking finding was that many of the episodes identified were very brief. In this paper the NPHS data were examined from a different perspective in order to derive a complementary perspective on the episode duration data. Data from the 1994/95 and 1996/97 cycles of the NPHS were used in the analysis. The longitudinal data were used to generate approximations of age and gender-specific incidence for members of the population over the age of 12 years. An estimate of prevalence was made from the 1996/97 cross-sectional file. A basic expression relating prevalence to incidence and mean duration of illness was then applied within age and gender categories. Taken together, the incidence and prevalence data from the NPHS suggest a longer duration than was indicated by the NPHS interview duration item. A probable explanation is that the NPHS duration question had an upper limit of 52 weeks, whereas some episodes of major depression last longer than this. Particularly long episodes could have a large impact on mean duration in the population. Nevertheless, these data confirm the heterogenous nature of this condition; many people with the syndrome of major depression may have quite brief episodes. PMID- 11397345 TI - Emigration patterns of cancer cases in Alberta, Canada. AB - Cancer registries are a unique source of data for population-based analysis of survival of cancer cases, but information on current vital status is essential. This paper describes a method to determine the last known vital status of cases and the emigration pattern of cancer cases diagnosed in Alberta. Data from the Alberta Cancer Registry (ACR) for the years 1985 1993 (83,446 cases), were linked to the Alberta Health Care Insurance Plan (AHCIP) registration file to identify cases that had left the province, and the date they emigrated. Ninety-nine percent of the ACR cases linked correctly to the AHCIP registration file. Three percent of cases had left Alberta by March 1998; For the first five years of follow-up between 0.6% and 0.8% of cases alive at the beginning of each year of follow-up left the province in the succeeding year. Seven percent of those diagnosed under 45 years of age left the province compared to less than 2% of those aged 65 and over. There was no difference in emigration patterns between the sexes. The cancer sites with good prognosis tended to have the highest proportion of emigrants. PMID- 11397346 TI - The economic burden of mental health problems in Canada. AB - This study provides a comprehensive estimate of the economic burden of mental health problems in Canada in 1998. In particular, it estimates the cost of non medical services that have not been previously published and the value of short term disability associated with mental health problems that were previously underestimated according to the definitions used here. The costs of consultations with psychologists and social workers not covered by public health insurance was $278 million, while the value of reduced productivity associated with depression and distress over the short term was $6 billion. Several data limitations suggest that these are underestimates. The estimated total burden of $14.4 billion places mental health problems among the costliest conditions in Canada. PMID- 11397347 TI - The storage of household long guns: the situation in Quebec. AB - This survey on the storage of household firearms in Quebec was conducted in 1994. At that time, 35% (175/504) of survey participants who kept long guns in their homes had failed to comply with Canadian firearm storage regulations. In most cases (85%; n=149), this was because at least one stored long gun was found to be both operable and accessible. Thirty-seven per cent of participants stated that no one, including themselves, had used their firearm(s) in the 12 months preceding the survey. These findings point to two possible ways of dealing with long guns kept in the home: render these weapons inoperable or inaccessible, which would increase the level of compliance with the regulations, and dispose of those no longer in use. The results of this survey have never been published before, and constitute the only information of this kind with respect to Quebec. PMID- 11397349 TI - Age and gender distributions of coronary artery calcium detected by electron beam tomography in 35,246 adults. AB - Electron beam tomography (EBT) is a noninvasive method used to detect coronary artery calcium (CAC). Due to the age-associated increase in incidence and magnitude of CAC, interpretation of results can be difficult. The purpose of this study was to develop a set of age- and gender-stratified CAC distributions to serve as standards for the clinical interpretation of EBT scans. Between 1993 and 1999, 35,246 asymptomatic subjects, 30 to 90 years of age, were self-referred for CAC screening using an Imatron EBT scanner. CAC score was calculated based on the number, areas, and peak computed tomographic density for each detected calcific lesion. CAC score in each coronary artery was equal to the sum of all lesions for that artery and the total CAC score was equal to the sum of the score of each artery. Total CAC scores were assigned to a percentile according to age and gender. CAC scores were reported at the 10th, 25th, 50th, 75th, and 90th percentiles for 16 age and/or gender groups. The prevalence of CAC increased with age for men and women. The extent of CAC differed significantly between men and women in the same age group. In summary, this study reports the distribution of CAC score by age and gender. Knowledge of the distribution of CAC, the effect of age on the total CAC score as well as the differences in total CAC scores that exist between men and women of similar age will assist the clinician in interpreting EBT CAC results. PMID- 11397350 TI - Relation of ST-segment changes in inferior leads during anterior wall acute myocardial infarction to length and occlusion site of the left anterior descending coronary artery. AB - We investigated the relation between left anterior descending (LAD) coronary artery morphology and inferior lead ST-segment changes to elucidate the clinical significance of such changes in 159 patients with anterior wall acute myocardial infarction (AMI). Patients with 1-vessel LAD artery lesions were divided into an ST depression group (n = 40), an ST elevation group (n = 25), and a no-ST-change group (n = 94) based on ST-segment changes in the inferior leads. The relation between each group and the infarct-related lesion and the presence of a wrapped LAD artery was then investigated. The percentage of patients with the infarct related lesion in the proximal LAD artery was significantly higher in the ST depression group and significantly lower in the ST elevation group. The percentage of patients with a wrapped LAD artery was significantly higher in the ST elevation group and significantly lower in the ST depression group. The wall motion index determined echocardiographically was significantly higher in the ST depression group and the no-ST-change group than in the ST elevation group. Our findings suggest that inferior lead ST-segment changes during anterior wall AMI arise as a result of competition between reciprocal changes caused by high lateral wall AMI due to lesions of the proximal LAD artery, which depress the ST segment, and inferoapical wall AMI due to a wrapped LAD artery, which elevates the ST segment. In patients with no ST-segment changes, echocardiography was useful for distinguishing the amount of affected LAD artery territory. PMID- 11397351 TI - Prediction of functional recovery in patients with chronic coronary artery disease and left ventricular dysfunction combining the evaluation of myocardial perfusion and of contractile reserve using nitrate-enhanced technetium-99m sestamibi gated single-photon emission computed tomography and dobutamine stress. AB - This study aimed to assess whether contractile reserve evaluation using dobutamine gated single-photon emission computed tomography (SPECT) improves the capability of quantitative perfusion analysis to predict functional recovery of viable hibernating myocardium. Resting and dobutamine nitrate-enhanced technetium 99m sestamibi (sestamibi) gated SPECT studies were performed in patients with coronary artery disease who had left ventricular dysfunction. Tracer activity was quantified, and wall motion and thickening visually scored. Reversible dysfunction was identified with gated SPECT repeated after coronary revascularization. Using the best activity threshold, perfusion quantification achieved 85% sensitivity and 55% specificity. Contractile reserve detection was significantly less sensitive (64%, p <0.0005), but more specific (88%, p <0.00001) than perfusion quantification. However, in the subgroup of hypokinetic segments, the sensitivity of contractile reserve assessment was just slightly lower than perfusion quantification (72% vs 91%, p = NS), whereas specificity was significantly higher (94% vs 23%, p <0.00001). Conversely, in the adyskinetic segments, perfusion quantification was significantly more sensitive than contractile reserve (82% vs 59%, p <0.005), but similarly specific (76% vs 85%, p = NS). Therefore, the identification of reversible dysfunction based on perfusion quantification in adyskinetic segments and on contractile reserve detection in hypokinetic segments was significantly more specific (83% vs 55%, p <0.00001) than standard quantitative perfusion SPECT, without major loss in sensitivity (78% vs 85%, p = NS). In conclusion, contractile reserve evaluation using dobutamine gated SPECT enhances the reliability of nitrate-enhanced sestamibi SPECT when used to predict reversible dysfunction in hypokinetic segments, whereas perfusion quantification remains superior in adyskinetic segments. PMID- 11397352 TI - Selective use of single-photon emission computed tomography myocardial perfusion imaging in a chest pain center. AB - Emergency department chest pain centers (CPCs) vary in their approach to patients with chest pain and nonischemic electrocardiograms (ECG). Although single-photon emission computed tomography (SPECT) myocardial perfusion imaging has been evaluated in this setting, both acutely at rest and after stress, we questioned its application in all patients. We prospectively evaluated the utility of selective SPECT imaging in a CPC (i.e., rest SPECT for ongoing pain, stress SPECT if unable to undergo exercise electrocardiography) and its impact on the overall disposition of all emergency department chest pain patients. Over 3 years, 2,601 patients were evaluated in a CPC (2,211 [85%] were sent home, 390 [15%] were hospitalized). Of 390 CPC patients hospitalized, 182 (47%) were diagnosed with coronary artery disease at the time of hospital discharge. Only 28 patients (1.1%) had an acute myocardial infarction. After 3 years, the proportion of all chest pain patients hospitalized and those diagnosed as "rule-out myocardial infarction" decreased from 53% to 41% and 32% to 18% of all chest pain patients, respectively (both p <0.0001). Overall, 906 patients (35%) required SPECT imaging to complete the CPC evaluation. Had SPECT imaging not been performed selectively, and all 906 patients been admitted, 762 (29%) would have been hospitalized unnecessarily based on the final diagnoses. Alternatively, sending all these patients home would have resulted in 144 (6%) inappropriate discharges of patients with coronary artery disease. A CPC protocol using the selective use of SPECT imaging permits the complete evaluation of all patients in the CPC, significantly reduces hospitalizations for chest pain, and restricts hospital admission to more appropriate patients. PMID- 11397353 TI - Creatine kinase-MB fraction elevation after percutaneous coronary intervention in patients with chronic renal failure. AB - We evaluated the short- and long-term clinical outcomes of 326 consecutive patients with chronic renal failure, not on dialysis, who had creatine kinase (CK)-myocardial band (MB) fraction elevation after successful percutaneous coronary intervention in a native coronary artery. Based on peak CK-MB levels measured after intervention, patients were divided into 3 groups: no elevation (group 1, n = 184), 1 to 3 x upper normal levels (group 2, n = 72), and >3 x upper normal levels (group 3, n = 70). Baseline clinical and angiographic characteristics were similar among the 3 groups. Angiographic success was similar among the 3 groups, although there was a significantly higher use of intra-aortic balloon pump in patients who had postprocedural CK-MB >3 x normal values and a higher rate of in-hospital complications, i.e., repeat catheterization, repeat target lesion intervention, pulmonary edema, renal function deterioration, emergency dialysis, and major bleeding complications. At 1-year follow-up, mortality rates were significantly higher in these patients (35.4% vs 22.0% for patients with CK-MB 1 to 3 x normal values and 16.7% for patients without CK-MB elevation, p = 0.007). Multivariate analysis showed that CK-MB >3 x normal (odds ratio 3.04; 95% confidence interval 1.41 to 6.57, p = 0.005) and intra-aortic balloon pump (odds ratio 1.49; confidence interval 1.15 to 1.93, p = 0.002) were independent predictors of late mortality. Therefore, patients with chronic renal failure who had CK-MB elevation >3 x the upper normal limit after a successful percutaneous coronary intervention had a higher incidence of in-hospital complications and a significantly higher mortality rate at 1-year follow-up than patients without CK-MB elevation or with <3 x normal CK-MB elevation. PMID- 11397354 TI - Correlation of polymorphisms to coagulation and biochemical risk factors for cardiovascular diseases. AB - Currently, the established risk factors for cardiovascular disease (CVD) are largely environmental in nature. Conflicting studies have suggested that mutations in specific coagulation genes may also provide a genetic basis for CVD risk. We reviewed clinical studies that examined the role of single nucleotide polymorphisms in coagulation and platelet factors, and a biochemical factor to determine if specific genotypes are correlated with patients with a history of arterial thrombotic diseases (acute coronary syndromes or stroke). A meta analysis was performed on studies for factors II (G20210A variant), V Leiden (G1691A), VII (R353Q), glycoprotein (GP) IIIa receptor (PI(A1/A2)), and methylenetetrahydrofolate reductase (MTHFR, C677T). There was no correlation for factor II or factor V polymorphisms to coronary artery disease (CAD) in 5,607 and 5,431 patients studied, respectively. There was also no correlation for factor II variants and stroke in 3,451 patients studied. For factor V, statistical significance was achieved for the G1691A variant on 3,399 patients with stroke (odds ratio [OR] 1.43, 95% confidence intervals [CI] 1.03 to 1.97). The GP IIIa PI(A1/A2) genotype was associated with increased risk for CAD in 7,920 patients (OR 1.12, 95% CI 1.01 to 1.24), but not for 1,855 patients who had a stroke (OR 0.80, 95% CI 0.62 to 1.04). The combined RQ and RR genotypes of factor VII R353Q were correlated to a reduced risk for CVD in 2,574 patients (OR 0.78, 95% CI 0.65 to 0.93), whereas the QQ genotype had offered more protection (OR 0.53, 95% CI 0.27 to 1.03). The TT homozygous variant of MTHFR was associated with CAD risk in 5,644 patients studied (OR 1.30, 95% CI 1.11 to 1.52) but not for 3,075 patients with stroke. This study shows that for some genes, further studies are unnecessary, whereas for others, no more enrollments are needed. The impact of certain genotypes must be examined in relation to other established risk factors and potentially new therapeutic strategies. PMID- 11397356 TI - Cardiac catheterization of low birth weight infants. AB - The increased survival of low birth weight infants means that more of these infants may be candidates for catheter interventions. There are few data on the results of cardiac catheterization in this group. This study aimed to analyze, retrospectively, cardiac catheterization of infants weighing < or =2.5 kg, with emphasis on the results of interventions. The complication rates of interventional and diagnostic procedures were compared. One hundred eleven catheterizations were performed in 107 patients between 1985 and 1998. Thirty-one procedures were interventional. Balloon atrial septostomy (n = 16), balloon pulmonary valvuloplasty (n = 10), balloon pulmonary angioplasty (n = 1), and coil occlusion of collateral vessels (n = 3) were all performed successfully. One infant (3%) died while undergoing myocardial biopsy. The reintervention rate for isolated pulmonary valve stenosis was 25% (2 of 8) at 1 month, 57% (4 of 7) at 6 months, and 71% (5 of 7) at 1 year. Complications were significantly more frequent during interventional (13 of 31, 42%) than during diagnostic (13 of 80, 16%) procedures. The most common complications during interventions were arrhythmias (3 of 31, 10%) and respiratory deterioration (3 of 31, 10%). Cardiac catheterization was technically feasible in all patients. Balloon pulmonary valvuloplasty and atrial septostomy provided good palliation in this patient group. The mortality of interventional procedures was low. The high incidence of respiratory complications suggests that low birth weight infants should undergo elective ventilation for interventional cardiac catheterization. PMID- 11397355 TI - Socioeconomic status as an independent risk factor for hospital readmission for heart failure. AB - The management of heart failure is characterized by high rates of hospital admission as well as rehospitalization after inpatient treatment of this disorder, whereas skillful medical care may reduce the risk of hospital admission. The purpose of this study was to examine the relation between income (as a measure of socioeconomic status) and the frequency of hospital readmission among a large and diverse group of persons treated for heart failure. We analyzed administrative discharge data from 236 nonfederal acute-care hospitals in New York State, involving 41,776 African-American or Caucasian hospital survivors with International Classification of Diseases, Ninth Revision, Clinical Modification codes for heart failure in the principal diagnosis position between January 1 and December 31, 1995. Household income was derived from postal ZIP codes and census data. We found that patients residing in lower income neighborhoods were more often women or African-Americans, had more comorbid illness, had higher use of Medicaid insurance, and were more often admitted to rural hospitals. There was a stepwise decrease in the crude frequency of readmission from the lowest quartile of income (23.2%) to the highest (20.0%) (p <0.0001 for Mantel-Haenszel chi-square test for trend across all quartiles; p <0.0001 for comparison between quartiles 1 and 4). After adjustment for baseline differences and process of care, income remained a significant predictor, with an increase in the risk of readmission noted in association with lower levels of income (adjusted odds ratio for quartile 1:4 comparison, 1.18; 95% confidence interval, 1.10 to 1.26, p <0.0001). We conclude that lower income patients hospitalized for treatment of heart failure in New York differ from higher income patients in important clinical and demographic comparisons. Even after adjustment for these fundamental differences and other potential confounding factors, lower income is a positive predictor of readmission risk. PMID- 11397357 TI - False lumen patency as a predictor of late outcome in aortic dissection. AB - Aortic dissection (AD) is a disease with a high-risk of mortality. Late deaths are often related to complications in nonoperated aortic segments. Between 1984 and 1996, we retrospectively analyzed the data of 109 patients with acute AD (81 men and 28 women; average age 61 +/- 14 years). All imaging examinations were reviewed, and a magnetic resonance imaging examination was performed at the time of the study. Aortic diameters were measured on each aortic segment. Predictive factors of mortality were determined by Cox's proportional hazard model, in univariate and multivariate analyses, using BMDP statistical software. Follow-up was an average of 44 +/- 46 months (range 24 to 164). Actuarial survival rates were 52%, 46%, and 37% at 1, 5, and 10 years, respectively, for type A AD versus 76%, 72%, and 46% for type B AD. Predictors of late mortality were age >70 years and postoperative false lumen patency of the thoracic descending aorta (RR 3.4, 95% confidence intervals 1.20 to 9.8). Descending aorta diameter was larger when false lumen was patent (31 vs 44 mm; p = 0.02) in type A AD. Furthermore, patency was less frequent in operated type A AD when surgery had been extended to the aortic arch. Thus, patency of descending aorta false lumen is responsible for progressive aortic dilation. In type A AD, open distal repair makes it possible to check the aortic arch and replace it when necessary, decreases the false lumen patency rate, and improves late survival. PMID- 11397358 TI - Heart failure in 2001: a prophecy revisited. PMID- 11397359 TI - Guidelines for interpretation of electron beam computed tomography calcium scores from the Dallas Heart Disease Prevention Project. PMID- 11397360 TI - The doctor in the courtroom. PMID- 11397361 TI - Comparison of six-month outcomes of percutaneous transluminal coronary angioplasty in patients > or =75 with those <75 years of age (the ROSETTA registry). PMID- 11397362 TI - Annual progression of coronary artery calcium by spiral computed tomography in hypertensive patients without myocardial ischemia but with prominent atherosclerotic risk factors, in patients with previous angina pectoris or healed acute myocardial infarction, and in patients with coronary events during follow up. PMID- 11397363 TI - Effect of selective serotonin reuptake inhibitors on platelets in patients with coronary artery disease. PMID- 11397364 TI - Relation between plasma fibrinogen level and coronary plaque morphology in patients with stable angina pectoris. PMID- 11397365 TI - Effect of acute psychological stress on the hypercoagulable state in subjects (spousal caregivers of patients with Alzheimer's disease) with coronary or cerebrovascular disease and/or systemic hypertension. PMID- 11397366 TI - Comparison of effects of losartan versus enalapril on fibrinolysis and coagulation in patients with acute myocardial infarction. PMID- 11397367 TI - Sex differences in uric acid and risk factors for coronary artery disease. PMID- 11397368 TI - Effect of quinapril versus nitrendipine on endothelial dysfunction in patients with systemic hypertension. PMID- 11397369 TI - Prevalence of heart failure in Asturias (a region in the north of Spain). PMID- 11397370 TI - Quantitative assessment of valve thickness in normal subjects by transesophageal echocardiography. PMID- 11397371 TI - Comparison of outcome in patients with culture-negative versus culture-positive active infective endocarditis. PMID- 11397372 TI - Comparison of myocardial perfusion single-photon emission computed tomography with coronary artery angiography after arterial switch operation. PMID- 11397373 TI - Auscultation versus echocardiography in a healthy population with precordial murmur. PMID- 11397374 TI - Use of contrast for image enhancement during stress echocardiography is cost effective and reduces additional diagnostic testing. PMID- 11397375 TI - John Hay and the earliest description of type II second-degree atrioventricular block. PMID- 11397377 TI - Evaluation of 1,3-butadiene, isoprene and chloroprene health risks. PMID- 11397378 TI - 1,3-Butadiene, isoprene and chloroprene: reviews by the IARC monographs programme, outstanding issues, and research priorities in epidemiology. AB - 1,3-Butadiene, isoprene and chloroprene have all been evaluated more than once by the IARC Monographs Programme on the Evaluation of Carcinogenic Risks to Humans, most recently in February 1998 (Volume 71). Summaries are available on-line at http://monographs.iarc.fr. 1,3-Butadiene is currently classified in Group 2A (probably carcinogenic to humans), on the basis of limited evidence for increased occupational cancer risk in humans plus sufficient evidence of carcinogenicity at multiple organ sites in rats and especially in mice exposed by inhalation. Four epidemiologic studies are available on cancer risk among workers exposed to 1,3 butadiene, one large study among styrene-butadiene rubber (SBR) workers, and one large and two small studies among 1,3-butadiene production workers. The results of the study of SBR workers suggest an association between butadiene exposure and leukaemia risk, which is consistent with the results of the large study of production workers. This latter study also suggested an increased risk of lymphoreticulosarcoma (ICD-8, 200). The major factors hampering the assessment of the available results are (i) possible misclassification of lymphoid and haematopoietic neoplasms, (ii) limitations in the assessment of past exposure (with the exception of the study of SBR workers) and (iii) a potential confounding effect of agents other than butadiene. Future research priorities include (i) the incorporation of newly developed biomarkers of exposure, (ii) the possible application of intermediate biomarkers, (iii) the replication of the study among SBR workers, possibly in Europe, and (iv) reanalysis of existing data in light of revisions of the classifications of leukaemias and lymphomas in the International Classification of Diseases for Oncology, Third Edition (2000). Isoprene is classified in Group 2B (possibly carcinogenic to humans), on the basis of sufficient evidence for carcinogenicity at multiple organ sites in both mice and rats, especially male mice, exposed by inhalation. No epidemiologic studies are available on cancer risk from occupational exposure to isoprene. Such studies could be conducted within the framework of existing or future studies of SBR workers, assuming that isoprene exposure can be disentangled from butadiene and styrene exposure. Chloroprene is classified in Group 2B on the basis of sufficient evidence for carcinogenicity at multiple organ sites in both mice and rats exposed by inhalation. Studies of chloroprene exposed workers now include chemical workers from the United States, China and Armenia as well as shoe workers from Russia. The results of the studies from China, Armenia and Russia suggest an excess risk of liver cancer. The risk of other neoplasms was not consistently increased. Limitations of available studies include possible bias from cohort enumeration, follow-up, and choice of reference population. In most studies the exposure assessment was poor, the possible confounding effect of co exposures was not addressed and the statistical power was low. The pathology of the cases of liver cancer should be reviewed. Future research priorities include a replication of available studies in well-defined populations and the development of biomarkers of exposure. PMID- 11397379 TI - Comparative carcinogenicity of 1,3-butadiene, isoprene, and chloroprene in rats and mice. AB - 1,3-Butadiene, isoprene (2-methyl-1,3-butadiene), and chloroprene (2-chloro-1,3 butadiene) are high-production-volume chemicals used mainly in the manufacture of synthetic rubber. Inhalation studies have demonstrated multiple organ tumorigenic effects with each of these chemicals in mice and rats. Sites of tumor induction by these epoxide-forming chemicals were compared to each other and to ethylene oxide, a chemical classified by the National Toxicology Program (NTP) and by the International Agency for Research on Cancer (IARC) as carcinogenic to humans. For this group of chemicals, there are substantial species differences in sites of neoplasia; neoplasia of the mammary gland is the only common tumorigenic effect in rats and mice. Within each species, there are several common sites of tumor induction; these include the hematopoietic system, circulatory system, lung, liver, forestomach, Harderian gland, and mammary gland in mice, and the mammary gland and possibly the brain, thyroid, testis, and kidney in rats. For studies in which individual animal data were available, mortality-adjusted tumor rates were calculated, and estimates were made of the shape of the exposure-response curves and ED10 values (i.e. exposure concentrations associated with an excess risk of 10% at each tumor site). Most tumorigenic effects reported here were consistent with linear or supralinear models. For chloroprene and butadiene, the most potent response was for the induction of lung neoplasms in female mice, with ED10 values of 0.3 ppm. Based on animal cancer data, isoprene and chloroprene are listed in the NTP's Report on Carcinogens (RoC) as reasonably anticipated to be a human carcinogen. Butadiene is listed in the RoC as known to be a human carcinogen 'based on sufficient evidence of carcinogenicity from studies in humans, including epidemiological and mechanistic information', with support from experimental studies in laboratory animals. Epidemiology data for isoprene and chloroprene are not considered adequate to evaluate the potential carcinogenicity of these agents in humans. PMID- 11397380 TI - A review of the epidemiology of 1,3-butadiene and chloroprene. AB - Butadiene epidemiologic research has focused primarily on one cohort of workers in the North American styrene-butadiene rubber (SBR) industry and on the largest cohort of workers in the United States butadiene monomer industry. The most recent studies of these populations are characterized by carefully enumerated study populations, extremely long and high quality mortality follow-up, accurate job categorizations, detailed exposure assessments, and comprehensive statistical analyses. Leukemia was clearly associated with increasing estimated butadiene exposure in the SBR study, but not in the monomer industry study. This has lead to hypotheses about exposure differences between these two industries and the presence of co-factors or confounders in the SBR industry. Research presented at this symposium should shed some light on these hypotheses. The chloroprene epidemiologic literature, on the other hand, is in an early stage of development. The existing studies are limited by poor exposure characterization, lack of control of potential confounding factors, incompleteness in cohort enumeration, short follow-up periods, and small numbers of cancer cases. The state of the science for chloroprene would be advanced by arranging more comprehensive studies than those that have been conducted to date. PMID- 11397381 TI - Species differences in the metabolism of olefins: implications for risk assessment. AB - Olefinic compounds are commercially valuable because they form useful polymeric substances. The same chemical property (presence of double bonds) that makes the olefins useful may also cause them to be toxic in the body. The double bonds of olefins can be oxidized by cytochrome P450 enzymes to epoxides, which are electrophiles that can react with DNA and may cause alterations in the genetic information carried by that macromolecule. Epoxides can be rendered inactive toward DNA by binding to proteins, by hydrolysis to diols through epoxide hydrolase enzymes (EHs), or by forming conjugates with glutathione via glutathione S-transferase (GST) activities. The balance between the oxidizing enzymatic activities and the hydrolyzing or conjugating enzymatic activities in the livers of different species can influence the potential toxicity of the olefins. The location of the enzymes and the potential for concerted reactions in which epoxides are inactivated immediately after formation will also influence the potential toxicity of the olefins. Cytochrome P450 enzymes and EHs are in microsomes located in the rough endoplasmic reticulum surrounding the nucleus where the DNA is located. GST is in the cytoplasm of the cell. In the case of 1,3 butadiene (BD), such enzymatic differences may strongly influence the toxicity in different species. The mouse, in which BD is a potent multi-site carcinogen, has the lowest microsomal EH activity of any species. This allows the monoepoxides formed in the microsomes by cytochrome P450 enzymes to be further oxidized to the highly genotoxic diepoxide (DEB), and both epoxides can either be released into the blood for distribution throughout the body or can react with DNA in the nucleus. The rat, in which BD is a weak carcinogen, has much higher levels of microsomal EH, and only trace amounts of DEB enter the bloodstream. Major BD metabolites in primates suggest that the hydrolysis pathway is even more prominent in primates than in rats. Data suggest that BD will be much less toxic in primates than in mice. Considering these quantitative differences in metabolism may help to reduce the uncertainties in extrapolating animal data on olefin toxicity to health risk assessments for humans exposed to the compounds. PMID- 11397382 TI - Genetic and reproductive toxicity of butadiene and isoprene. AB - Butadiene (BD) and its 2-methyl analogue, isoprene, have been extensively studied in animals and BD in population studies. Both chemicals are metabolised by liver cytochrome P450 dependent monogenases to monoepoxide and diepoxide intermediates. The diepoxide intermediates of both compounds were mutagenic in Salmonella typhimurium. However, unlike the monoepoxide of BD, the monoepoxides of isoprene were not mutagenic. It appears that they have no alkylating capacity. BD did not induce somatic cell mutation and recombination or sex-linked recessive lethal mutation in Drosophila melanogaster and isoprene produced no increase in chromosomal aberrations in CHO cells in vitro. Comparative concentrations of haemoglobin adducts in the blood of mice and rats after exposure to BD indicated that reaction with blood may decrease the levels of reactive intermediates available to tissues in rats, but not in mice contributing to greater potency of BD in the mouse. For isoprene, the adducts reach approximately the same concentrations in both species. DNA adducts have also been detected in testicular and lung cells of mice after BD exposure. The level of epoxybutene haemoglobin adducts was significantly elevated in BD-exposed workers, but lower than in rats and mice. In conjunction with the toxicology and carcinogenesis studies for BD and isoprene, additional mice were included for the evaluation of cytogenetic effects. Both chemicals produced increases in sister chromatid exchanges in bone marrow cells and in the frequency of micronuclei in normochromatic and polychromatic erythrocytes, but only BD produced an increase in the percent of bone marrow cells with chromosomal aberrations. At similar doses, the effects with BD were 2-3 times larger than with isoprene. There were also increased hprt mutation frequencies in rats and mice after BD exposure. Biomonitoring studies with hprt mutations in lymphocytes showed conflicting results, with both positive and negative findings. BD has been shown to be positive in one human cytogenetic biomonitoring study and not in three others, but chromosomal aberrations were increased in BD-exposed workers after challenge with gamma rays. Re-analysis of GSTTI null individuals showed positive results. There was an increase in spermatid micronuclei in mice by BD and its metabolites and in rats only by its metabolites. The cytotoxic response of germ cells in mice is greater than in rats. Dominant lethal mutations have been induced by BD and diepoxybutane, but not by epoxybutene. There was some evidence of congenital malformations in mice after BD exposure and there was a linear concentration-related induction of heritable translocations in mice. There was no induction of dominant lethal mutations or congenital malformations in rats. Using the heritable translocation data in mice, it has been determined that if a worker is continually exposed over 5 or 6 weeks to 20-25 ppm of BD, the risk of producing a child with a balanced reciprocal translocation is twice as high as the background risk. Since genetic damage cannot be measured directly in human germ cells, risk to such cells can also be estimated from germ cells and somatic cells of the mouse and human somatic cells using the parallelogram approach. Using doubling doses, the fourth corner of the parallelogram was calculated as a doubling dose for human germ cells of 4390 ppm/h. However, it is still questioned if man is more like rat than mouse in terms of sensitivity to exposure. Similar germ cell data do not exist for isoprene. In conventional developmental studies, where rats and mice were exposed to BD, maternal toxicity was shown in rats but there was no evidence of developmental toxicity or teratogenic effects and there was a small effect on sperm morphology. After exposure to isoprene, there was no adverse effect on rat dams or other reproductive indices. In mice, there was reduced foetal body weight and decreased maternal weight gain and isoprene also affected ovarian follicles. There was a reduction in testicular function parameters such as testicular weight and sperm motility. PMID- 11397383 TI - Overview of the acute, subchronic, reproductive, developmental and genetic toxicology of beta-chloroprene. AB - beta-Chloroprene (CD), the 2-chloro derivative of 1,3-butadiene, is used for the manufacture of the synthetic rubber, polychloroprene. Acute inhalation studies show that CD is lethal to Crl:CD rats at >2300 p.p.m. (4 h); the primary target organ effects were pulmonary hemorrhage and edema, and hepatic necrosis. In 2- and 4-week inhalation studies in Fischer 344 (F344) and Wistar rats, early deaths occurred at 500 and > or =161 p.p.m., respectively. Organ system injury was found in the nose (degeneration/metaplasia of olfactory epithelium), liver (centrilobular necrosis), and blood (decreased red blood cell count in F344 rats only). In a 90-day inhalation study with F344 rats, degeneration/metaplasia of the olfactory epithelium and reduced nonprotein sulfhydryl content of lungs and liver were found in animals exposed to 80 p.p.m., and anemia, hepatocellular necrosis, and forestomach inflammation were observed at 200 p.p.m. In a 90-day study with B6C3F1 mice, CD caused deaths at 200 p.p.m., the highest concentration tested, and epithelial hyperplasia of the forestomach at 80 p.p.m. Other than a slight (<10%) reduction in sperm motility in male rats at 200 p.p.m., all other reproductive parameters (sperm count or morphology in males, and estrous cyclicity or cycle length in females) were unaffected in these 90-day rat/mouse studies. There were no significant indications of neurological toxicity. The study No-Observable Adverse Effect Level was 32 p.p.m. based on nasal injury in rats. Despite some early reports of reproductive system abnormalities at levels <1 p.p.m., recent studies show no embryotoxic or developmental toxicity in female Wistar or Crl:CD rats, or in New Zealand White rabbits at CD exposure concentrations up to 25 or 175 p.p.m., respectively. In a one-generation reproduction study with Wistar rats, CD produced growth retardation in the F(0) generation exposed to 100 p.p.m., and in the F(1) offspring at 33 and 100 p.p.m.; no effects on reproductive parameters or histopathology were found. CD is nonmutagenic in standard plate incorporation bacterial reverse mutation assays (Ames assays) but positive using direct gas-phase incubation methods. Bacterial mutagenicity (primarily base pair substitution) was either negative or weakly positive when freshly prepared CD was tested. Mutagenicity increased markedly with time, presumably from CD dimer formation, and also by addition of liver S9 metabolic activation system. In vivo micronucleus, chromosome aberration and sister chromatid exchange studies in mice showed no structural chromosomal damage. Overall, the pathological effects in the liver and nose dominate the subchronic toxicity of CD. The genotoxicity of CD is inconsistent and requires further study. PMID- 11397384 TI - Butadiene--progress under the European Union Existing Substances Regulation. AB - A draft risk assessment report (RAR) for 1,3-butadiene has been produced by the UK under the European Union's Existing Substances Regulation (ESR). In line with the requirements of ESR, the RAR presents an evaluation of the hazards and risks of butadiene to human health (for workers, consumers and the general public) and to the environment. If as a result of the risk assessment, concerns for human health and/or the environment are identified, proposals for measures to reduce those risks may be developed. In relation to the human health elements of the ESR RAR for butadiene, the risk assessment involves a comparison of health effects against exposures in the three population sectors. The position arrived at on risk is described in qualitative rather than quantitative terms. For butadiene, the conclusion reached for the human health hazard assessment has been influenced significantly by both the animal and human data available. Species differences in toxicity, in particular with regard to mutagenicity and carcinogenicity, are considered along with the mechanistic arguments for these differences and their relevance for human health. The information presented in the RAR has been extensively discussed within the EU and a generally agreed position has been reached, with respect to human health hazard and risk. It is concluded that whilst the risks to human health at current exposure levels in the workplace, for consumers and following exposure via the environment are uncertain, there are concerns for human health effects for all three populations. Thus, there is a need to limit the risks. The next stage in the ESR process will be to consider what, if any, additional risk reduction measures may be required, taking into account existing control measures. PMID- 11397386 TI - Significance of 1,3-butadiene to the US air toxics regulatory effort. AB - Because of its prevalence, particularly as a combustion by-product, 1,3-butadiene is a particularly important air toxic. It plays a significant role in all air toxics regulatory efforts in the US. The various requirements of the Federal Clean Air Act (CAA) dealing with air toxics are reviewed and the significance of 1,3-butadiene in each area is discussed in light of what is known about its emissions and health effects. The impacts of the changes in the understanding of 1,3-butadiene cancer potency over the past 15 years demonstrates the possible impact of such benchmarks and the importance of using the best science in understanding public health risks. PMID- 11397385 TI - Health risk assessment of 1,3-butadiene as a Priority Substance in Canada. AB - 1,3-Butadiene was included in the second list of Priority Substances to be assessed under the Canadian Environmental Protection Act. Potential hazards to human health were characterized on the basis of critical examination of available data on health effects in experimental animals and occupationally exposed human populations, as well as information on mode of action. Based on consideration of all relevant data identified as of April 1998, butadiene was considered highly likely to be carcinogenic to humans, and likely to be a somatic and germ cell genotoxicant in humans. In addition, butadiene may also be a reproductive toxicant in humans. Estimates of the potency of butadiene to induce these effects have been derived on the basis of quantitation of observed exposure-response relationships for the purposes of characterization of risk to the general population in Canada exposed to butadiene in the ambient environment. PMID- 11397387 TI - BD monomer and elastomer production processes. AB - The monomer 1,3 butadiene (BD) is a product of the petrochemical industry. It is used to make several elastomers including the very high volume styrene butadiene rubber (SBR) that comprises the bulk of automobile tires. It is also used to make polybutadiene rubber that is used in parts of tires, coatings, composites and other products. The monomer can be converted to chlorobutadiene (chloroprene) and used to make polychloroprene (neoprene). BD is one of the several olefins created by cracking hydrocarbons in the presence of steam. A mixed C4 stream from the steam cracker is then sent to a BD monomer extraction unit. Modern units typically use dimethyl formamide as the extraction solvent. SBR is commonly made by the copolymerization of BD and styrene, along with various additives to control the reaction, in a water emulsion. The reaction proceeds in a continuous chain of reactors until it is 'shortstopped' by a strong reducing agent. After removing unreacted monomers from the stabilized latex, it is blended, coagulated and dewatered. The resulting dry rubber crumb is bailed, film wrapped and stored in crates. The polymerization of BD to make polybutadiene rubber can be conducted as a water suspension type polymerization similar to SBR or in a solvent system followed by solvent recovery and transfer into water suspension. PMID- 11397388 TI - Manufacture and use of chloroprene monomer. AB - There are only six manufacturers of chloroprene (CD) located outside of the former Communist Bloc countries. Five of these manufacture chloroprene from butadiene via a two step process consisting of chlorination and subsequent dehydrochlorination. The sixth dimerizes acetylene and then hydrochlorinates the dimer to produce CD. Both the acetylene and butadiene-based manufacturing processes are conducted within sealed systems. Therefore, the only potential exposure during manufacture is sampling for product quality, strainer/filter changing and accidental leaks or spills. All six manufacturers produce CD only for the production of polymer. Manufacturers convert CD into polychloroprene polymer (PCP) via emulsion polymerization. A solution of CD and other ingredients is mixed with an aqueous caustic solution to produce an emulsion. Free radicals initiate polymerization and free radical scavengers are added to stop the reaction at the desired conversion. After unreacted monomer is removed, PCP can be sold as a latex (colloid) or isolated and dried to produce solid product. The PCP manufacturing process is also designed to be a closed system. However, the extreme reactivity of CD and the inherent stickiness of PCP polymer, makes it necessary to 'open' the process to remove polymer. This results in potential for operator exposure, in addition to the items mentioned for CD manufacture. With the use of appropriate engineering controls, work practices and personal protective equipment for activities with exposure potential, actual workplace exposures are controlled below the occupational exposure limits (2-10 ppm). The solid PCP products, which comprise approximately 92% of the total amount of polymer manufactured, contain <1 ppm CD. Most PCP latexes contain <0.1% residual CD. So, the potential for CD industrial exposure is limited to CD and PCP manufacture. PMID- 11397389 TI - Overview of isoprene monomer and polyisoprene production processes. AB - Isoprene monomer is employed in the manufacture of polyisoprene and various copolymers (with isobutylene, butadiene, styrene, other monomers). Recent surveys of air levels of isoprene in US monomer and polymer manufacturing facilities have demonstrated TWA concentrations <1 ppm in 90% of collected samples, and 98% were <10 ppm. Tumorigenic responses have been observed in isoprene-exposed rodents. However, the relevance of these findings to health in occupational environments can only be assessed following completion of additional and ongoing mechanistic research. PMID- 11397390 TI - Ambient concentrations of 1,3-butadiene in the UK. AB - This paper assesses the current knowledge of 1,3-butadiene as an atmospheric pollutant, considers measurement techniques and reviews available data on 1,3 butadiene monitoring and emissions estimates. Atmospheric chemistry, sources of emission, current legislation, measurement techniques and monitoring programmes for 1,3-butadiene are reviewed. There have been comparatively few studies of the products of oxidation of 1,3-butadiene in the atmosphere. However, on the basis of the available information, and by analogy with the oxidation mechanism for the widely-studied and structurally similar natural hydrocarbon isoprene (2-methyl 1,3-butadiene), it is possible to define some features of the likely oxidation pathways for 1,3-butadiene. The total UK 1,3-butadiene emission to the atmosphere for 1996 has been estimated at 10.60 kTonnes. 1,3-Butadiene is a product of petrol and diesel combustion; consequently this total is dominated by road transport exhaust emissions (accounting for some 68% of the total). Off-road vehicles and machinery are responsible for 14% of the total UK emission. 1,3 Butadiene is used in the manufacture of numerous rubber compounds, and consequently emissions arise from both the manufacture and use of 1,3-butadiene in industrial processes. Emissions from the chemical industry account for 18% of the UK total emission- 8% from 1,3-butadiene manufacture and 10% from 1,3 butadiene use. The United Kingdom Expert Panel on Air Quality Standards (EPAQS) has published a report on 1,3-butadiene, and recommended a national air quality standard of 1.0 ppb (expressed as an annual rolling mean). This was adopted by the Government as part of the National Air Quality Strategy (NAQS) in 1997, and a target of compliance by 2005 was set. Work conducted for the review of the NAQS (1999) indicated that it was likely that all locations would be compliant with the national standard by the end of 2003. As a result, the review updated the air quality objective for 1,3-butadiene, with the deadline for compliance being brought forward to 31/12/2003. The UK Hydrocarbon Monitoring Network provides continuous hourly measurements of 1,3-butadiene at 13 sites, and has been operational since 1993. The dataset that is available allows spatial and temporal trends to be evaluated, and has proved to be invaluable in characterising the current ambient levels of 1,3-butadiene in the UK. Hourly maximum concentrations of 1,3-butadiene of up to 10 ppb (1 ppb=1 ppb, i.e. 1 vol. of 1,3-butadiene in 1,000,000,000 vol. of air. 1 ppb of 1,3-butadiene is ca. equal to 2.25 microg m( 3) at 20 degrees C) may be measured for several hours at the sites. Monthly mean concentrations are typically 0.1-0.4 ppbv. At most sites, these levels are driven by emissions from motor vehicles. Occasionally emissions of 1,3-butadiene from industrial sources may elevate 1,3-butadiene concentrations to several tens of ppb. Trend analysis of the data suggests that ambient concentrations of 1,3 butadiene in the UK are declining at about 10% per year. PMID- 11397391 TI - Occupational exposure to butadiene, isoprene and chloroprene. AB - Workers are exposed to butadiene, isoprene and chloroprene in the manufacture of these monomers and in their use in the production of various elastomers. These include styrene butadiene rubber, polybutadiene, polyisoprene, butyl rubber and neoprene. Monomer production and extraction are done in typical closed chemical process units where low background levels of the monomers are the result of minor leaks in valves and pumps. Occasionally, higher levels occur as a result of planned or unplanned events that cause releases. Polymer production is also a closed process, but the occasional clogging of pipes and equipment with polymer requiring maintenance operations where some release is likely occurs much more often than for monomer production. For this reason, exposure levels are generally higher on polymer production units. Polymer finishing is essentially an open process, but almost all monomer should have been stripped from the polymer before finishing. Where small amounts of solvents or monomers remain in the polymer and are volatilized in finishing, they are captured by vapor control systems. As a result, exposures in finishing are typically low. Measured levels of exposure in recent years are presented. In general, modern levels of exposure are well below OSHA, ACGIH and other applicable limits. Few measurements were made prior to the 1970s, but epidemiological estimates made by modeling suggest that levels could have been quite high in the 1940s and 1950s. In these years, manual reactor cleaning was common, and pumps often leaked. PMID- 11397392 TI - Human exposures to monomers resulting from consumer contact with polymers. AB - Many consumer products are composed completely, or in part, of polymeric materials. Direct or indirect human contact results in potential exposures to monomers as a result of migrations of trace amounts from the polymeric matrix into foods, into the skin or other bodily surfaces. Typically, residual monomer levels in these polymers are <100 p.p.m., and represent exposures well below those observable in traditional toxicity testing. These product applications thus require alternative methods for evaluating health risks relating to monomer exposures. A typical approach includes: (a) assessment of potential human contacts for specific polymer uses; (b) utilization of data from toxicity testing of pure monomers, e.g. cancer bioassay results; and (c) mathematical risk assessment methods. Exposure potentials are measured in one of two analytical procedures: (1) migration of monomer from polymer into a simulant solvent (e.g. alcohol, acidic water, vegetable oil) appropriate for the intended use of the product (e.g. beer cans, food jars, packaging adhesive, dairy hose); or (2) total monomer content of the polymer, providing worse-case values for migratable monomer. Application of toxicity data typically involves NOEL or benchmark values for non-cancer endpoints, or tumorigenicity potencies for monomers demonstrated to be carcinogens. Risk assessments provide exposure 'safety margin' ratios between levels that: (1) are projected to be safe according to toxicity information, and (2) are potential monomer exposures posed by the intended use of the consumer product. This paper includes an example of a health risk assessment for a chewing gum polymer for which exposures to trace levels of butadiene monomer occur. PMID- 11397393 TI - Metabolism and molecular toxicology of isoprene. AB - Isoprene (2-methylbuta-1,3-diene) is a large-scale petrochemical used principally in the manufacture of synthetic rubbers. It is also produced by plants and trees and is the major endogenous hydrocarbon formed by mammals, probably from mevalonic acid. Isoprene is metabolised by mammals in processes that involve epoxidation by cytochrome P450-dependent monooxygenases to the isomeric mono epoxides, (1-methylethenyl)-oxirane and 2-ethenyl-2-methyloxirane. Further metabolism of the mono-epoxides to mutagenic isoprene di-epoxides, (2, 2')-2 methylbioxiranes, can also occur. The oxidations to the mono- and di-epoxides occur enantioselectively and diastereoselectively. The mono-epoxides are hydrolysed enantioselectively to vicinal diols under catalysis by epoxide hydrolase. 2-Ethenyl-2-methyloxirane is also readily hydrolysed non enzymatically. Because of the stereochemical possibilities for metabolites, the metabolism of isoprene is complex. The metabolism of isoprene by liver microsomes in vitro from a range of species including rat, mouse and human shows significant differences between species, strains and gender in respect of the diastereoselectivity and enantioselectivity of the metabolic oxidation and hydrolysis reactions. The impact of the extra methyl in isoprene on di-epoxide reactivity also appears to be critically important for the resulting biological effects. Isoprene di-epoxides may exhibit a lower cross-linking potential in vivo compared to butadiene di-epoxides. Differences in metabolism and reactivity of metabolites may be factors contributing to the significant differences in toxicological response to isoprene observed between species. PMID- 11397394 TI - Cellular and molecular basis for species, sex and tissue differences in 1,3 butadiene metabolism. AB - Species differences in 1,3-butadiene (BD) bioactivation and detoxication have been implicated in the greater sensitivity of mice to the carcinogenic effects of BD compared to rats, but the molecular basis for species differences in BD metabolism is not well understood. Previous and recent work conducted in this laboratory has examined the relative rates of BD oxidation to epoxybutene (EB) in male and female B6C3F1 mouse tissues, characterized the major cytochrome P450 enzymes involved in BD bioactivation in these tissues, and determined the potential utility of the freshly isolated hepatocyte model to investigate species differences in metabolism of BD and related compounds. Collectively, the results suggest a role for P450s 2E1, 2A5, and 4B1 in sex and tissue differences in BD bioactivation in the mouse. When coordinated metabolism of EB was investigated in male B6C3F1 mouse and Sprague-Dawley rat hepatocytes, the hepatocytes from both species were found to catalyze EB oxidation to meso- and (+/-)-diepoxybutane (DEB), EB hydrolysis to 3-butene-1,2-diol (BDD), and EB conjugation to form GSH conjugates (GSEB). The metabolite area under the curve (AUC) exhibited dependence on the EB concentration used. However, the EB activation/detoxication ratios with the mouse hepatocytes were much higher than the ratios obtained with the rat hepatocytes. These results illustrate the potential utility of the hepatocyte model for estimating flux through competing metabolic pathways and predicting in vivo metabolism of EB. Collectively, the results may allow a better understanding of the molecular and kinetic basis of species differences in BD metabolism and may lead to a more accurate assessment of human risk. PMID- 11397395 TI - First-pass metabolism of 1,3-butadiene in once-through perfused livers of rats and mice. AB - First-pass metabolism of 1,3-butadiene (BD) leading to 1,2-epoxy-3-butene (EB), 1,2:3,4-diepoxybutane (DEB), 3-butene-1,2-diol (B-diol), 3,4-epoxy-1,2-butanediol (EBD) and crotonaldehyde (CA) was studied quantitatively in the once-through BD perfused liver of mouse and rat by means of an all-glass gas-tight perfusion system. Metabolites were analyzed using gas chromatography equipped with mass selective detection. The perfusate consisted of Krebs-Henseleit buffer (pH 7.4) containing bovine erythrocytes (40%v/v) and BD. The perfusion flow rates through the livers were 3-4 ml/min (mouse) and 17-20 ml/min (rat). The BD concentrations in the liver perfusates were 330 nmol/ml (mouse) and 240 nmol/ml (rat) being high enough to reach almost saturation of BD metabolism. The mean rates of BD transformation were about 0.014 and 0.055 mmol/h per liver of a mouse and a rat, respectively, being similar to the values expected from in-vivo measurements. There were marked species differences in the formation of BD metabolites. In the effluent of mouse livers, all three epoxides (EB: 9.4 nmol/ml; DEB: 0.06 nmol/ml; EBD: 0.07 nmol/ml) and B-diol (8.2 nmol/ml) were detected. In the perfusate leaving naive rat livers, only EB and B-diol were found. In that of rat liver, EB concentration was 8.5 times smaller than in that of mouse liver, whereas B-diol concentrations were similar in the effluent liver perfusate of both species. CA was below the limit of its detection (60 nmol/l) in the liver perfusate of mice and of naive rats. Of BD metabolized, the sum of the metabolites investigated in the effluent amounted to only 30% (mouse) and 20% (rat). In first experiments with rat liver, glutathione (GSH) was depleted by pretreating the animals with diethylmaleate. With the exception of EBD (not quantifiable due to an interfering peak), all other metabolites including CA were found in the effluent perfusate summing up to about 70 and 100% of BD metabolized, which indicates the quantitative importance of the GSH dependent metabolism. In summary, the results demonstrate the relevance of an intrahepatic first-pass metabolism for metabolic intermediates of BD, which undergo further transformation immediately after their production in the liver before leaving this organ. Hitherto, the occurrence of this first-pass metabolism was only hypothesized. The findings will help to explain the drastic species difference between mice and rats in the carcinogenic potency of BD. PMID- 11397396 TI - The metabolism of beta-chloroprene: preliminary in-vitro studies using liver microsomes. AB - Based on analogy with butadiene and isoprene, the metabolism of beta-chloroprene (2-chloro-1,3-butadiene, CD) to reactive intermediates is likely to be a key determinant of tumor development in laboratory rodents exposed to CD by inhalation. The purpose of this study is to identify species differences in toxic metabolite (epoxide) formation and detoxification in rodents and humans. The in vitro metabolism of CD was studied in liver microsomes of B6C3F1 mice, Fischer/344 and Wistar rats, Syrian hamsters, and humans. Microsomal oxidation of CD in the presence of NADP(+), extraction with diethyl ether, and analysis by GC mass selective detection (MSD) indicated that (1-chloroethenyl)oxirane (CEO) was an important metabolite of CD in the liver microsomal suspensions of all species studied. Other potential water-soluble oxidative metabolites may have been present. The oxidation of CD was inhibited by 4-methyl pyrazole, an inhibitor of CYP 2E1. CEO was sufficiently volatile at 37 degrees C for vial headspace analysis using GC-MSD single ion monitoring (m/z=39). CEO was synthesized and used to conduct partition measurements along with CD and further explore CEO metabolism in liver microsomes and cytosol. The liquid-to-air partition coefficients for CD and CEO in the microsomal suspensions were 0.7 and 58, respectively. Apparent species differences in the uptake of CEO by microsomal hydrolysis were hamster approximately human>rats>mice. Hydrolysis was inhibited by 1,1,1-trichloropropene oxide, a competitive inhibitor of epoxide hydrolase. A preliminary experiment indicated that the uptake of CEO in liver cytosol by GSH conjugation was hamster>rats approximately mice (human cytosol not yet tested). In general, the results suggest that metabolism may help explain species differences showing a greater sensitivity for CD-induced tumorigenicity in mice, for example, compared with hamsters. Additional experiments are in progress to quantify the kinetic parameters of CD oxidation and CEO metabolism by enzymatic hydrolysis and conjugation by glutathione S-transferase for in cytosol. A future goal is to use the kinetic rates to parameterize a physiologically based toxicokinetic model and relate the burden of toxic metabolite to the cancer dose response observed in experimental animals. PMID- 11397397 TI - Physiological modeling of butadiene disposition in mice and rats. AB - The earliest physiological models of 1,3-butadiene disposition reproduced uptake of the gas from closed chambers but over-predicted steady-state circulating concentrations of the mutagenic intermediates 1,2-epoxybut-3-ene and 1,2:3,4 diepoxybutane. A preliminary model based on the observation of a transient complex between cytochrome P450 and microsomal epoxide hydrolase on the endoplasmic reticulum membrane reproduced the blood epoxide concentrations as well as the chamber uptake data. This model was enhanced by the addition of equations for the production and detoxication of 3,4-epoxybutane-1,2-diol in the liver, lungs, and kidneys. The model includes flow-restricted delivery of butadiene and its metabolites to compartments for lungs, liver, fat, kidneys, gastrointestinal tract, other rapidly perfused tissues, and other slowly perfused tissues. Blood was distributed among compartments for arterial, venous, and tissue capillary spaces. Channeling of the three bound epoxides to epoxide hydrolase and their release from the endoplasmic reticulum are competing processes in this model. Parameters were estimated to fit data for chamber uptake of butadiene and epoxybutene, steady-state blood concentrations of epoxybutene and diepoxybutane, and the fractions of the inhaled dose of butadiene that appears as various excreted metabolites. The optimal values of the apparent K(m)s of membrane-bound epoxides for epoxide hydrolase were only 5% of the values for the cytosolic substrate, consistent with the observation of a transient complex between epoxide hydrolase and the cytochrome P450 that produces the epoxide. This proximity effect corresponds to the notion that epoxides produced in situ have privileged access to epoxide hydrolase. The model also predicts considerable accumulation of epoxybutanediol, in agreement with the observation that most of the DNA adducts in animals exposed to butadiene arise from this metabolite. PMID- 11397398 TI - Development of a preliminary physiologically based toxicokinetic (PBTK) model for 1,3-butadiene risk assessment. AB - Potential health effects of human exposure to 1,3-butadiene (BD) are of concern due to the use of BD in industry and its low-level presence throughout the environment. Physiologically based toxicokinetic (PBTK) models of BD in rodents have been developed by multiple research groups in an effort to explain species differences in toxicity (especially carcinogenic potency) through toxicokinetics. PBTK modeling of dose metrics related to a non-cancer endpoint, ovotoxicity in experimental animals, was conducted. The cumulative area under the blood concentration vs. time curve (AUC) for the metabolite diepoxybutane (butadiene diepoxide, DEB) was found to be consistent with ovotoxicity in mice and rats exposed to BD by inhalation or epoxybutene (butadiene monoepoxide, EB) or DEB by intraperitoneal injection. This suggests that cumulative DEB AUC may also be an appropriate metric for possible human risk. A preliminary human PBTK model was assembled for the eventual assessment of reproductive risk to humans and for prioritizing the determination of model parameters. The preliminary model accurately predicted published data on exhaled breath BD concentrations in a human volunteer exposed to BD by inhalation. The fit was relatively insensitive to the rate constant for BD epoxidation. Sensitivity analyses were conducted on this human PBTK model. Using a range of published rate constants, human blood DEB was found to be sensitive to rates of epoxidation of EB to DEB and hydrolysis of EB and DEB, but not BD epoxidation. Because of the large ranges of rates measured in vitro for these reactions, different combinations of in-vitro rates produce varying predictions of blood DEB concentration. Thus, validation of a human PBTK model with human biomonitoring data will be essential to produce a PBTK model that can be applied to risk assessment. PMID- 11397399 TI - Mutational spectrum of 1,3-butadiene and metabolites 1,2-epoxybutene and 1,2,3,4 diepoxybutane to assess mutagenic mechanisms. AB - 1,3-Butadiene (BD) is a multisite carcinogen and is mutagenic in multiple tissues of B6C3F1 mice. BD is bioactivated to at least three directly mutagenic metabolites: 1,2-epoxybutene (EB), 1,2-epoxy-3,4-butanediol (EBD), and 1,2,3,4 diepoxybutane (DEB). However, the contribution of these individual metabolites to the carcinogenicity and in vivo mutatidnal spectrum of BD is uncertain. To assess the role of two BD metabolites EB and DEB in the in vivo mutagenicity of the parent compound BD, we examined the in vitro mutational spectra of EB and DEB in human and rodent cells. We also examined the in vivo mutagenicity and mutational spectrum of inhaled EB in the lung. In the bone marrow and spleen of B6C3F1 laci transgenic mice, BD-induced an increased frequency of the identical class of point mutations at A:T base pairs: AT-->GC transitions and AT-->TA transversions. BD exposure also induced an increased frequency of GC-->AT transitions in the spleen that was not observed in bone marrow, demonstrating tissue-specific differences in mutation spectrum. Exposure of Rat2 laci transgenic cells and human TK6 lymphoblasts to EB-induced an increased frequency of AT-->TA transversions. DEB exposure induced an increased frequency of AT-->TA transversions and partial deletions at hprt in human cells. In Rat laci transgenic cells, DEB was not mutagenic at laci but induced an increased frequency of micronuclei. In contrast to inhaled BD, inhaled DEB and EB were not mutagenic in the bone marrow or spleen. However, EB was mutagenic in the lungs. In the lung of mice, EB-induced specific increases in GC-->AT transitions, AT- >TA transversions, and deletion events. AT-->TA transversions are the most consistent mutation observed across biological systems following in vivo exposure to BD or in vitro exposures to EB and DEB. Although, BD exposure in mice induces chromosomal alterations and single base substitutions, the specific BD metabolite that induces the genetic events leading to tumors is uncertain. At present, it appears that only DEB can effectively induce this range of mutagenic events at levels of this metabolite that occur in the blood of mice exposed to BD. Detailed investigations to identify relevant biomarkers of BD exposure and response, particularly DNA adducts or lesions, that can be biologically linked to the range of genotoxic events known to occur in mice exposed to BD are needed. PMID- 11397400 TI - Mutagenicity at the Hprt locus in T cells of female mice following inhalation exposures to low levels of 1,3-butadiene. AB - A study was conducted to test the hypothesis that repeated low level exposures to 1,3-butadiene (BD), approaching the OSHA occupational threshold for this chemical, produce a significant mutagenic response in mice. Female B6C3F1 mice (4 5 weeks of age) were exposed by inhalation for 2 weeks (6 h/day, 5 days/week) to 0 or 3 ppm BD, and then necropsied at 4 weeks after the cessation of exposures to measure the frequency of mutations (MF) at the Hprt locus using the T-lymphocyte clonal assay. At necropsy, T cells were isolated from spleen and cultured in the presence of mitogen, growth factors, and a selection agent. Cells were scored for growth on days 8-9 after plating to determine cloning efficiencies (CEs) and Hprt MFs. There was a marginal but significant reduction in the growth of splenic T cells from mice exposed to 3 ppm (n=27) compared with control mice (n=24) (P=0.004), suggesting the occurrence of BD-induced cytotoxicity at this low exposure concentration. In addition, the average Hprt MF in mice exposed to 3 ppm BD [1.54+/-0.82 (S.D.)x10(-6)] was significantly increased by 1.6-fold over the average control value of 0.96+/-0.51 (S.D.)x10(-6) (P=0.004). Comparisons of these data to earlier Hprt mutagenicity studies of mice exposed to high concentrations of BD (where significant mutagenic but not cytotoxic effects were observed) indicate that the ability to detect the cytotoxic and mutagenic responses of T cells to low levels of BD was enhanced by using a much larger sample size than usual for both the control and treatment groups. Additional analyses of the quantitative relationships between CE and MF demonstrated that CE had no significant effect upon MF values in sham-exposed control mice or mice exposed to low-level BD. Furthermore, the approaches for assessing the impact of CE and clonality on Hprt MFs in these control and BD-exposed mice were applied with the same rigor as in in vivo Hprt mutagenicity studies in human children. The overall study results support the conclusion that short-term low-level BD exposure is mutagenic in the mouse. PMID- 11397401 TI - N7-guanine adducts of the epoxy metabolites of 1,3-butadiene in mice lung. AB - Epoxy metabolites of 1,3-butadiene are electrophilic and can bind to nucleophilic sites in DNA forming DNA adducts. In this study, guanine N7 adducts of epoxy butene and guanine N7 adducts of epoxy butanediol were measured in lung tissues of mice inhalation exposed to various concentrations of 1,3-butadiene. 32P postlabeling of DNA adducts were used to demonstrate that the DNA adducts derived from epoxybutene and epoxybutanediol were formed in a dose dependent manner. More than 98% of all adducts detected were formed from epoxybutanediol. Enantiomeric distribution of the adducts formed in vivo differs from that of in vitro experiments demonstrated before. In the case of epoxybutene most of the adducts were formed to the terminal carbon of the S-epoxybutene enantiomer. Most of the adducts derived from epoxybutanediol were formed from the 2S-3R enantiomer. The data demonstrates that enzymatic processes involved with activation and/or detoxification of the metabolites are enantiospecific and/or DNA repair machinery repairs the damage with stereochemical considerations. These are the crucial factors if interspecies differences in tumor sensitiveness is concerned. PMID- 11397402 TI - Point mutations of K-ras and H-ras genes in forestomach neoplasms from control B6C3F1 mice and following exposure to 1,3-butadiene, isoprene or chloroprene for up to 2-years. AB - 1,3 Butadiene (BD), isoprene (IP) and chloroprene (CP) are structural analogs. There were significantly increased incidences of forestomach neoplasms in B6C3F1 mice exposed to BD, IP or CP by inhalation for up to 2-years. The present study was designed to characterize genetic alterations in K- and H-ras proto-oncogenes in a total of 52 spontaneous and chemically induced forestomach neoplasms. ras mutations were identified by restriction fragment length polymorphism, single strand conformational polymorphism analysis, and cycle sequencing of PCR amplified DNA isolated from paraffin-embedded forestomach neoplasms. A higher frequency of K- and H-ras mutations was identified in BD-, IP- and CP-induced forestomach neoplasms (83, 70 and 57%, respectively, or combined 31/41, 76%) when compared to spontaneous forestomach neoplasms (4/11, 36%). Also a high frequency of H-ras codon 61 CAA-->CTA transversions (10/41, 24%) was detected in chemically induced forestomach neoplasms, but none were present in the spontaneous forestomach neoplasms examined. Furthermore, an increased frequency (treated 13/41, 32% versus untreated 1/11, 9%) of GGC-->CGC transversion at K-ras codon 13 was seen in BD-, and IP-induced forestomach neoplasms, similar to the predominant K-ras mutation pattern observed in BD-induced mouse lung neoplasms. These data suggest that the epoxide intermediates of the structurally related chemicals (BD, IP, and CP) may cause DNA damage in K-ras and H-ras proto-oncogenes of B6C3F1 mice following inhalation exposure and that mutational activation of these genes may be critical events in the pathogenesis of forestomach neoplasms induced in the B6C3F1 mouse. PMID- 11397403 TI - Using DNA and hemoglobin adducts to improve the risk assessment of butadiene. AB - The purpose of this paper is to review what we know about various biomarkers of butadiene in animal, human and in vitro studies, and to draw inferences from these data that impact on the accurate assessment of human risks for cancer. Studies comparing the DNA and hemoglobin adducts of butadiene with exposure, metabolism and genotoxicity have provided a great deal of insight that is applicable to biologically based risk assessment. First, the DNA and hemoglobin adduct data strongly support the conclusion that 3,4-epoxy-1,2-butanediol is the major electrophile available for binding to these macromolecules. Biomarker studies have also provided insight into the possibility of a sensitive population associated with the GSTT1 null genotype. While it is clear that lymphocytes from GSTT1 null individuals are more sensitive for the induction of sister chromatid exchanges (SCE) following in vitro exposure to 1,2,3,4-diepoxybutane, there was no such increase in SCE or other biomarkers of genotoxicity in workers exposed to 1-3 p.p.m. butadiene, regardless of GST genotype. The globin adduct data also demonstrate that there is roughly a tenfold range for interindividual differences in the metabolism of butadiene. This type of analysis represents an excellent means for providing scientific data for this critical determinant. Another useful application of hemoglobin adducts in risk assessment was demonstrated by regressing data for various endpoints for genotoxicity against that individual's biologically effective dose, thereby providing an independent mechanism for evaluation that excludes any possible confounding by inappropriate controls. Finally, biomarker studies have identified critical gaps in our knowledge that are needed for the accurate assessment of butadiene. Most notable of these is the lack of diepoxide-specific biomarkers in mice, rats and humans. PMID- 11397404 TI - Genetic and dietary factors affecting human metabolism of 1,3-butadiene. AB - The objective of this project was to determine the factors associated with differences in butadiene (BD) inhalation uptake and the rate of metabolism for BD to epoxy butene by monitoring exhaled breath during and after a brief exposure to BD in human volunteers. A total of 133 subjects (equal males and females; four racial groups) provided final data. Volunteers gave informed consent and completed a questionnaire including diet and alcohol use. A venous blood sample was collected for genotyping CYP2E1. Subjects received a 20 min exposure to 2.0 ppm of BD, followed by a 40 min washout period. The total administered dose was 0.6 ppm*h, which is in the range of everyday exposures. Ten, 1 or 2 min exhaled breath samples (five during and five after exposure) were collected using an optimized strategy. BD was determined by GC-FID analysis. Breathing activity (minute ventilation, breath frequency and tidal volume) was measured to estimate alveolar ventilation. After the washout period, 250 mg of chlorzoxazone were administered and urine samples collected for 6 h to measure 2E1 phenotype. The total BD uptake during exposure (inhaled BD minus exhaled) was estimated. A three compartment PBPK model was fitted to each subject's breath measurements to estimate personal and population model parameters, including in-vivo BD metabolic rate. A hierarchical Bayesian PBPK model was fit by Monte Carlo simulations to estimate model parameters. Regression and ANOVA analyses were performed. Earlier data analysis showed wide ranges for both total uptake BD and metabolic rate. Both varied significantly by sex and age, and showed suggestive differences by race, with Asians having the highest rates. The analyses reported here found no correlation between total BD uptake and metabolic rate. No significant differences were found for oxidation rates by 2E1 genotype or phenotype, but the rates showed trends consistent with reported differences by genotype and phenotype for chlorzoxazone metabolism. No effects on metabolic rate were observed for long-term alcohol consumption, or consumption in the past 24 h. Overall, neither dietary factors nor genetic differences explained much of the wide variability in metabolic rates. Population characteristics, age, sex, and race, were the most important explanatory variables, but a large fraction of the total variability in metabolism remains to be explained. PMID- 11397405 TI - Biomarkers for assessing occupational exposures to 1,3-butadiene. AB - The overall objective of this study was to evaluate a continuum of biomarkers in blood and urine for their sensitivities as indicators of low level occupational exposures to 1,3 butadiene (BD). The study design was largely cross-sectional, with biological samples collected within a short timeframe. Personal 8-h BD exposure measures were made on several occasions over a 60-day period for each potentially exposed worker in order provide maximum accuracy for this independent variable and to accommodate the different expression intervals of the several biomarkers. Co-exposures to styrene, toluene and benzene were also measured. The study included 24 BD monomer production workers (mean BD exposure=0.642 mg/m(3)), 34 polymerization workers (mean BD exposure=1.794 mg/m(3)) and 25 controls (mean BD exposure=0.023 mg/m(3)). The several biomarkers were measured by a consortium of investigators at different locations in the US and Europe. These biomarkers included: (1) metabolic genotypes (CYP2E1, EH, GST M1, GST T1, ADH2, ADH3), determined in Prague and Burlington, VT; (2) urinary M1 and M2 metabolites (1,2 dihydroxy-4-[N-acetylcysteinyl]-butane and 1-hydroxy-2-[N-acetylcysteinyl]-3 butene, respectively), determined in Albuquerque, NM and Leiden; (3) hemoglobin adducts (N-[2-dihydroxy-3-butenyl]valine=HBVal and N-[2,3,4 trihydroxybutyl]valine=THBVal), determined in Amsterdam and Chapel Hill, NC, respectively; (4) HPRT mutations determined by autoradiographic assay in Galveston, TX, with slides re-read in Burlington, VT; (6) hypoxanthine-guanine phosphoribosyltransferase (HPRT) mutations determined by cloning assay in Leiden with mutational spectra characterized in Burlington, VT; (7) sister chromatid exchanges and chromosome aberrations determined by standard methods and FISH analysis in Prague. Urinary M1 and M2 metabolites and HBVal and THBVal hemoglobin adducts were all significantly correlated with BD exposure levels, with adducts being the most highly associated. No significant relationships were observed between BD exposures and HPRT mutations or any of the cytogenetic endpoints, regardless of method of assay. PMID- 11397406 TI - Markers for carcinogenicity among butadiene-polymer workers in China. AB - We examined a spectrum of genotoxic and other outcomes in 41 butadiene-polymer production workers and 38 nonexposed controls, in China, to explore the role of butadiene in human carcinogenesis. Among butadiene-exposed workers, median air exposure was 2 ppm (6-h TWA), due largely to intermittent high-level exposures. Compared to unexposed subjects, butadiene-exposed workers had greater levels of hemoglobin N-(2,3,4-trihydroxybutyl)valine (THBVal) adducts (P<0.0001), and adduct levels tended to correlate, among butadiene-exposed workers, with air measures (P=0.03). Butadiene-exposed workers did not differ, however, from unexposed workers with respect to frequency of uninduced or diepoxybutane-induced sister chromatid exchanges, aneuploidy as measured by fluorescence in situ hybridization of chromosomes 1, 7, 8 and 12, glycophorin A variants or lymphocyte hprt somatic mutation. Also among the exposed, greater THBVal levels were not associated with increases in uninduced sister chromatid exchanges, aneuploidy, glycophorin A, or hprt mutations. Butadiene-exposed workers had greater lymphocyte (P=0.002) and platelet counts (P=0.07) and lymphocytes as a percent of white blood cells were moderately correlated with greater THBVal levels (Spearman's rho=0.32, P=0.07). Among butadiene-exposed workers, several serum cytokines correlated with THBVal adduct levels. Overall, the study demonstrated exposure to butadiene in these workers, by a variety of short-term and long-term measures, but did not show specific genotoxic effects, at the chromosomal or gene levels, related to that exposure. PMID- 11397407 TI - Assessment of butadiene exposure in synthetic rubber manufacturing workers in Texas using frequencies of hprt mutant lymphocytes as a biomarker. AB - 1,3-Butadiene (BD), which is used to manufacture synthetic rubber, is a mutagen and carcinogen. Because past occupational exposures have been associated with an increased risk of leukemia, there has been a dramatic reduction in workplace exposure standards. The health benefits of these reduced levels of occupational exposure to BD will be difficult to evaluate using relatively insensitive traditional epidemiological studies; however, biomarkers can be used to determine whether there are genotoxic effects associated with recent exposures to BD. In past studies of BD-exposed workers in Southeast Texas, we observed an increase in the frequency of lymphocytes with mutations in a reporter gene, hprt. Frequencies of hprt mutant cells correlated with air levels of BD and with the concentration of a BD metabolite in urine. Average exposures to 1-3 parts per million (p.p.m.) of BD were associated with a threefold increase in hprt variant (mutant) frequencies (Vfs). We now report results from a follow-up study of workers in a synthetic rubber plant in Southeast Texas. Thirty-seven workers were evaluated on three occasions over a 2-week period for exposure to BD by the use of personal organic vapor monitors and by determining the concentration of a BD metabolite in urine. The frequency of hprt mutants was determined, by autoradiography, with lymphocyte samples collected 2 weeks after the final exposure measurement. Based on their work locations, the study participants were assigned to high-exposure (N=22) or low-exposure (N=15) groups. The BD exposure, +/-standard error, of the workers in the high-exposure group (1.65+/-0.52 p.p.m.) was significantly greater than the low-exposure group (0.07+/-0.03 p.p.m.; P<0.01). The frequency of hprt mutant lymphocytes was also significantly different in the two groups (high, 10.67+/-1.5 x 10(-6); low, 3.54+/-0.6 x 10(-6); P<0.001). The concentration of the urine metabolite was greater in the high-exposure group, but the difference was not significant. The correlation coefficient between hprt Vf and BD exposure levels was r=0.44 (CI(95), 0.11-0.69; P=0.011). This study reproduced the findings from a previous study at this plant. Although studies of butadiene exposed workers in other countries have not detected an effect of exposure on frequencies of hprt mutant lymphocytes, we have repeatedly observed this result in our studies in Texas. PMID- 11397408 TI - Cohort studies of chloroprene-exposed workers in Russia. AB - Two retrospective cohort studies were conducted to assess the risk of cancer among workers exposed to chloroprene (2-chloro-1,3-butadiene) (CP). One is a study of incidence and mortality among 2314 production workers employed in the CP production plant in Yerevan, Armenia, between 1940 and 1988. The cohort was followed up for cancer incidence for the years 1979-1990 and for cancer mortality for 1979-1988. The second study is a mortality study among 5185 shoe manufacturing workers in Moscow who used polychloroprene latex and glue. Shoe workers were employed between 1940 and 1976, and followed from 1979 through 1993. The standardized incidence ratios (SIR) and standardized mortality ratios (SMR) were calculated using the Armenian and Moscow population as reference. An internal comparison analysis based on Poisson regression modeling was conducted. In the Yerenan cohort, incidence and mortality from all cancers were below expectation, but increased incidence (SIR, 3.27; 95% confidence interval (CI), 1.47-7.27), and mortality (SMR, 3.39; CI, 1.09-10.5) from liver cancer were noted. A dose-response relationship was suggested between the risk of liver cancer and indices of CP exposure. For the entire Moscow cohort, all-cause mortality was close to expectation and all-cancer mortality was increased. There was an increase in the mortality from liver cancer (SMR, 2.4; CI, 1.1-4.3), kidney cancer (SMR, 1.8; CI, 0.9-3.4), and leukemia (SMR, 1.9; CI, 1.0-3.3). Mortality from liver cancer and leukemia was associated with various indicators of CP exposure. A similar, although less consistent, pattern was found for kidney cancer. The association between CP exposure and risk of leukemia may be due to concomitant exposure to benzene. The results for liver cancer point towards a carcinogenic effect of CP. PMID- 11397409 TI - A cohort study of workers exposed to chloroprene in the department of Isere, France. AB - A cohort study that comprised 533 men was carried out to evaluate the risk of cancer incidence among workers exposed to chloroprene between 1966 and 1997 in a production plant in Isere (France). The risk of cancer was evaluated for the period 1979-1997 from data recorded by the cancer registry in the department of Isere. Standardised incidence ratios (SIR) were calculated using the whole population of the department as reference. An excess of risk of all malignant neoplasms in the cohort was shown (SIR, 1.26; CI, 0.88-1.77). This excess concerned cancers of the lung (SIR, 1.84; CI, 0.84-3.49) and of the head and neck (SIR, 1.89; CI, 0.87-3.59). Where lung cancer was concerned, there was an increase in risk depending on the length of exposure. This relationship was inverse for the head and neck. The excess of risk of lung cancer was the highest in people not exposed or only slightly exposed. For head and neck cancer, this excess was the highest in people with medium exposure. Our results did not confirm the excess of risk of liver cancer discovered during other cohort studies aiming to evaluate the role of chloroprene. The variations in the excess of the risk of cancer of the lung, head and neck were hardly compatible with a professional risk factor. PMID- 11397410 TI - Leukemia and exposure to 1,3-butadiene, styrene and dimethyldithiocarbamate among workers in the synthetic rubber industry. AB - This study evaluated relations between exposure to 1,3-butadiene (BD), styrene (STY) and dimethyldithiocarbamate (DMDTC) and mortality from leukemia among synthetic rubber industry workers. Subjects were 13130 men employed for at least 1 year during 1943-1991 at any of six plants that manufactured synthetic rubber. Death certificates and medical records identified workers with leukemia. Cumulative exposure estimates were based on plant- and time period-specific process and task characteristics, linked to subjects' work histories. Poisson regression estimated relative rates (RRs) for workers exposed to each agent compared to unexposed workers. Leukemia (N=59) was positively associated with BD ppm-years (RRs of 1.0, 1.2, 2.0 and 3.8, for exposures of 0, >0-<86.3, 86.3 <362.2 and 362.2+ ppm-years; only the RR for the highest exposure category was statistically significant), STY ppm-years (RRs of 1.0, 1.2, 2.3 and 3.2, for exposures of 0, >0-<20.6, 20.6-<60.4 and 60.4+ ppm-years; only the RR for the highest exposure category was statistically significant) and DMDTC mg-years/cm (RRs of 1.0, 2.3, 4.9 and 2.9, for 0, >0-<566.6, 566.6-<1395.1 and 1395.1+ mg years/cm; the RR for each non-zero exposure category was statistically significant) after adjusting for age and years since hire. After further adjusting each agent-specific set of RRs for the other two agents, a positive but imprecise relation remained for BD and DMDTC but not for STY. The association with BD was stronger for ppm-years due to exposure intensities >100 ppm than for ppm-years due to lower concentrations. BD and DMDTC, but not STY, were positively associated with leukemia in multivariable analyses. The independent effect of each agent was difficult to evaluate because of correlations with other agents and imprecision. PMID- 11397412 TI - A mortality, morbidity, and hematology study of petrochemical employees potentially exposed to 1,3-butadiene monomer. AB - This three-part study is an update of a previous report that examined the mortality, morbidity, and hematological data of employees from a petrochemical facility in Texas who had potential exposure to 1,3-butadiene monomer. The first part describes the updated cause-specific mortality of 614 workers. Vital status for each cohort member was ascertained through 1998, a 9-year extension of the previous study. The second part is an examination of the morbidity experience of cohort members who were still working during 1992-1998, including 289 of the 614 mortality cohort members. The third part is an evaluation of the hematological results from routine health surveillance and/or medical examinations. Approximately 430 of the 614 employees who had complete blood count (CBC) data as of December 1999 were included in the hematological evaluations. The most recent examination containing CBC data was used and compared with similar data for over 2600 other employees from this facility. Overall mortality during the follow-up period, 1948-1998, was significantly lower than for the local comparison population (standardized mortality ratio (SMR) of 0.55 with a 95% confidence interval (CI) of 0.42-0.70). Mortality for all cancer was also significantly lower (SMR, 0.57; 95% CI, 0.32-0.92). Mortality for all lymphohematopoietic cancer was about the same as the comparison population (SMR, 1.06; 95% CI, 0.22 3.11). None of the cause-specific morbidity was in excess compared with an internal comparison group. There were no differences in the distribution or mean values of hematological variables (e.g. white blood cells, red blood cells, hemoglobin, platelets, mean corpuscular volume) between the butadiene cohort and the comparison group, or between a subgroup of workers in jobs with the highest potential for butadiene exposure (i.e. shipping) and the rest of the butadiene cohort. The findings of this study suggest that the butadiene exposure at this facility in the last 20 years does not pose a health hazard to employees. PMID- 11397411 TI - A cohort mortality study among workers at a 1,3 butadiene facility. AB - This is a cohort mortality study of 2800 male workers employed at least 6 months between 1943 and 1996 at a 1,3-butadiene monomer production facility. Earlier analyses of mortality for this cohort found statistically significant deficits for the 'all causes of death' category, and a lower than expected mortality for most leading causes of death. Past analyses also showed a significant elevation for deaths from cancers of the lymphohematopoietic system that was mainly due to an increase in deaths from lymphosarcoma. The purpose of this update was to examine the patterns of mortality through the end of 1999, for five additional years of follow-up. Persons who had become eligible between the last cohort update and April 1996 were added. Cohort membership was closed after April 1996 due to the sale of the facility. A total of 1422 deaths through December 1999 were identified, giving over 200 more deaths than in the last report on this aging cohort. The standardized mortality ratio (SMR) for all causes of death was 89 [95% confidence interval (95% CI) =84-94], which is statistically significantly low, and that for all malignant neoplasms was 90 (95% CI=81-101). The SMR for all lymphohematopoietic cancers (LHC) was 141 (95% CI=105-186) and is statistically significant. The SMR for leukemia was 129 (95% CI=77-204) and for non-Hodgkin's lymphoma (NHL), 148 (95% CI=89-231). The LHC elevations again were found only in workers first employed before 1950 and, in the group with the highest potential for exposure to butadiene, the elevations were highest in the short-term workers. Survival analyses were performed for all LHC [International Classification of Diseases (ICD) codes 200-209], NHL (ICD codes 200, 202), and leukemia (ICD codes 204-207) using an estimate of cumulative butadiene exposure as a time-dependent explanatory variable defined as a combination of job exposure class, calendar time, and length of time in job. The relative risks for the above causes of death were essentially 1.0, suggesting that there was no increase in risk with increasing butadiene exposure. PMID- 11397413 TI - Application of process chemistry and SAR modelling to the evaluation of health findings of lower olefins. AB - Epidemiology studies show increased leukemia mortality among styrene butadiene rubber (SBR) workers but not among butadiene monomer production employees. A detailed review of the SBR manufacturing process indicates that sodium dimethyldithiocarbamate (DMDTC) introduced into the SBR manufacturing process for a period in the 1950s coincides with increased leukemia mortality. Using the Computer-Optimized Molecular Parametric Analysis of Chemical Toxicity (COMPACT), we assessed the enzyme (cytochrome P450) substrate specificity of an olefin series including 1,3-butadiene (BD) and also modeled its interaction with DMDTC. These analyses showed correlation of a structural/electronic parameter--the COMPACT radius--with the presence or absence of cytogenetic activity and also found that DMDTC would inhibit the oxidative metabolism of BD at least at high concentrations. Both DMDTC and its diethyl analog (DEDTC) bind with CYP 2E1 and CYP 2A6. Both of these isoforms are important in the initial oxidative metabolism of butadiene and other olefins. In co-exposure studies in mice of DMDTC with BD or with epoxybutene (EB), we found that there was a reduced increase in genotoxic activity based on micronuclei induction compared with BD or EB exposure alone. Treatment with DMDTC significantly increased the protein carbonyl contents of hepatic microsomes compared with that of controls, a finding that may be related to DMDTC's activity as a prooxidant. Co-exposure with DMDTC and EB increased hepatic microsomal carbonyls to levels significantly greater than those of DMDTC treated mice, while EB administration in the absence of DMDTC did not change protein carbonyls relative to those of controls. The increase in hepatic microsomal protein carbonyls suggests that DMDTC may modulate EB metabolism towards the formation of reactive intermediates that react with proteins. The present molecular modeling and mechanistic studies suggest that co-exposure of BD and DMDTC is a plausible biological hypothesis regarding increased leukemia risk among SBR workers. PMID- 11397414 TI - The influence of co-exposure to dimethyldithiocarbamate on butadiene metabolism. AB - Treatment of rats and mice with a single oral dose of dimethyldithiocarbamate (DMDTC; 250 mg/kg) had a marked effect on hepatic CYP2E1 and aldehyde dehydrogenase activities, measured in vitro, for up to 24 h after dosing. The same treatment did not affect CYP2A6, glutathione S-transferase, epoxide hydrolase, alcohol dehydrogenase activities or hepatic glutathione levels. As a consequence of the loss of CYP2E1 activity, butadiene metabolism in liver fractions from DMDTC treated rats and mice was markedly reduced, as was the metabolism of the mono-epoxide to the di-epoxide in mouse liver. The conversion of the mono-epoxide to the diol by epoxide hydrolases was not affected by DMDTC treatment. Urinary excretion of radioactivity, following dosing with DMDTC and exposure to 200 ppm C-14 butadiene for 6 h, was markedly reduced in rats, but increased in mice. The profiles of urinary metabolites were qualitatively similar from mice exposed to butadiene to those exposed after dosing with DMDTC. In the rat, pre-dosing with DMDTC resulted in the formation of three additional urinary metabolites following exposure to butadiene. Overall, DMDTC appears to impact qualitatively and quantitatively on the metabolism of butadiene. The nature and full significance of these changes has yet to be characterised. PMID- 11397415 TI - Insights into the toxicokinetics and toxicodynamics of 1,3-butadiene. AB - 1,3 Butadiene (BD) is a colorless gas used in the production of synthetic rubber and plastics. BD is carcinogenic in rats and mice, however, there are striking species differences in cancer potency and spectrum of tumors, with mice being more susceptible to tumor induction than rats. Epidemiology studies suggest an excess incidence of leukemia in workers in the styrene-butadiene rubber industry. Consideration of mechanisms of BD carcinogenicity can provide insights into differences in cancer potency between rodents and serve to elucidate the extent to which BD exposure may cause cancer in humans. Mechanistic research in the areas of biochemical toxicology, molecular biology, molecular dosimetry, and susceptibility factors can impact BD cancer risk assessment for humans. This research has focused on quantitating species differences in the metabolism of BD and BD epoxides, defining molecular lesions produced by BD epoxides, identifying biomarkers for BD exposure to explore metabolic pathways in humans, and determining potential risk factors for sensitive subpopulations. BD is activated by P450 isozymes, including CYP2E1, to at least two genotoxic metabolites, epoxybutene (EB) and diepoxybutane (DEB). Dosimetry data from several laboratories on EB and DEB following inhalation exposure to BD indicate that blood concentrations of EB were four-eight-fold higher in mice compared with rats and that blood concentrations of DEB were 25-100-fold higher in mice than in rats. The higher levels of these two DNA-reactive metabolites in mice compared with rats probably contribute to the species differences in carcinogenic effects of BD between mice and rats. In vitro metabolism studies of BD in rats, mice, and human tissues indicate that there are significant quantitative species differences in the metabolic activation of BD to EB and DEB and the detoxication of EB and DEB. Activation/detoxication ratios calculated using in vitro kinetic constants reveal that ratios in mice were greater than in both rats and humans. In vitro data are consistent with in vivo dosimetry data and cancer potency for rodents, and suggest that humans may be at a decreased risk. Data on mutagenicity and mutational spectra of BD epoxides show mechanistic differences between EB- and DEB-induced mutational events suggesting involvement of DEB in the development of cancer. Concentrations of DEB that are genotoxic in vitro are within the range of concentrations measured in mice in vivo, whereas concentrations of EB that are genotoxic in vitro are ten-100-fold greater than concentrations observed in vivo. Characterization of molecular events indicate that EB-induced genotoxicity is due to point mutations and small deletions, while DEB induces point mutations, small deletions, and large-scale deletions involving many base pairs. The extent to which epoxybutanediol is involved in BD carcinogenesis is not known. Molecular dosimetry studies in rodents and humans have focused on urinary metabolites and DNA and hemoglobin adducts. Data from these studies are consistent with in vivo and in vitro metabolism data providing further support for the differences in metabolic activation and deactivation of BD and BD epoxides across species and the role of DEB in tumor development. Research on potential susceptibility factors points to other P450 isozymes, in addition to CYP2E1, that are involved in both the metabolic activation and mutagenicity of BD. Taken together, mechanistic data on BD toxicokinetics and toxicodynamics provide an integrated insight into critical steps in initiation of cancer, metabolites responsible for cancer, sensitive biomarkers for exposure, and potential risk factors for individual susceptibility. Available evidence suggests that BD is unlikely to be a human carcinogen at the low exposure concentrations currently encountered in the environment or workplace. PMID- 11397416 TI - Comparative toxicity of dithiocarbamates and butadiene metabolites in human lymphoid and bone marrow cells. AB - Apparent differences in the pattern of leukemia risk have been observed between workers employed in 1,3-butadiene (BD) monomer production and those working in styrene-butadiene rubber production (SBR). There are a number of possible explanations for these discrepancies, including differences in disease classification and diagnosis as well as possible quantitative and qualitative differences in occupational exposure between these two industries. This led us to evaluate the possibility that the pattern of disease observed in SBR might be influenced by the presence of an important class of biologically reactive chemicals, dithiocarbamates (DTC), that were present in SBR but not BD monomer production. Therefore, we compared the immunotoxic and hematotoxic activities of DTC and BD metabolites in human immune and hematopoietic cells. Relative to the mouse, human CD34+ bone marrow cells are relatively resistant to the direct effects of BD metabolites, with only the bis-oxide producing any evidence of suppression of clonogenic response at concentrations between 1 and 10 microM. Similarly, treatment of human CD4+ lymphocytes with known (2,3-epoxybutene) and putative BD metabolites (D,L-butane-bis-oxide, (2S,3R)-3-epoxybutane-1,2-diol) does not result in appreciable T-cell toxicity at concentrations likely to be encountered in vivo. In contrast, treatment of human cells with DTC at concentrations as low as 100 nM results in significant suppression of hematopoietic clonogenic response and T-lymphocyte function. Additional studies in our laboratory and others suggest a role for copper in DTC toxicity in both human lymphocytes and bone marrow cells, although the pattern of altered transcriptional regulation observed is markedly different in these two cell populations. These results are consistent with the pattern of DTC toxicity previously observed in clinical and molecular studies. PMID- 11397417 TI - Future research needs for the health assessment of 1,3-butadiene. AB - In recent years, several organizations have evaluated the health effects of 1,3 butadiene. Dissimilar conclusions about the carcinogenicity of 1,3-butadiene have been reached, due to differences in interpretation of the same data. Although 1,3 butadiene has been extensively studied, various uncertainties and data gaps remain which will require further clarification to reduce the uncertainties in future health assessments. This paper discusses future research needs respective of cancer and non-cancer effects that would be useful for future assessments as to how 1,3-butadiene affects health. PMID- 11397418 TI - Dose-response implications of the University of Alabama study of lymphohematopoietic cancer among workers exposed to 1,3-butadiene and styrene in the synthetic rubber industry. AB - New quantitative cancer risk estimates for exposure to 1,3-butadiene are presented. These estimates are based on the most recent human epidemiologic data developed by Drs Delzell and Macaluso and their colleagues at the University of Alabama at Birmingham. The implications of Poisson regression analyses of the relative rate for leukemia are explored using their updated dose estimates and lymphohematopoietic cancer data. The Poisson regression model in these analyses has the same form as in the U.S. Environmental Protection Agency (EPA)'s draft risk assessment of 1,3-butadiene [U.S. Environmental Protection Agency, Health Risk Assessment of 1,3-Butadiene - External Review Draft, National Center for Environmental Assessment, Office of Research and Development, 63 Fed. Reg. 7167 (February 12, 1998) Publication NCEA-W-0267, Washington, 1998]. Consistent with the proposed cancer risk assessment guidelines of the EPA and the EPA's draft risk assessment, the exploration includes the maximum likelihood estimate of the 'effective concentration' (EC(01)) corresponding to an extra risk of leukemia of 0.01 (1%) from a lifetime continuous exposure to 1,3-butadiene based on a linear dose-response model and the cumulative 1,3-butadiene dose metric (ppm-years). The incorporation of the most recent exposure estimates results in a 2.5-fold decrease in the estimates of leukemia risks computed by EPA. In addition, three changes proposed by the American Chemistry Council (formerly the Chemical Manufacturers Association) to the EPA's Science Advisory Board (SAB) for EPA's draft risk assessment of 1,3-butadiene are incorporated into the calculation. This results in approximately an additional fivefold decrease in the risk estimates of leukemia. The leukemia cancer risk estimates in the EPA's draft risk assessment of 1,3-butadiene decrease by approximately a factor of 13-fold when the updated epidemiologic data and the alternative numbers proposed by industry to the SAB are both incorporated. Specifically, the maximum likelihood estimate of the EC(01) increases from EPA's 1.2 ppm to 2.8 ppm on the basis of the updated epidemiologic data and increases further to 15.1 ppm when the CMA's proposed changes are also incorporated. PMID- 11397419 TI - The classification of lymphomas and leukemias. AB - Classifications of lymphomas and leukemias have developed from two distinct clinical needs - to understand the natural history of these diseases in order to predict outcome and to make treatment decisions in a rational fashion. The utility of classifications for research on etiology of these diseases has not guided their development in the past. The classification of leukemias and lymphomas has undergone dramatic changes with increasing understanding of the development of the normal immune cells. Historically, the first entity to be recognised was Hodgkin disease. Other malignancies of the lymphatic system were then called 'Non-Hodgkin Lymphomas' (NHL), a distinction that remains valid even today. Cancers that tend not to form distinct masses but usually present with a raised white blood cell count were called leukemias. As knowledge has improved, however, the early juxtaposition of leukemias versus lymphomas has lost relevance, since often the same entity can present in either way. With better understanding the terms 'lymphosarcoma' and 'reticulosarcoma', which were earlier widely applied have been replaced by more precise terminology. Different classifications have been put forward over the years. The 'Revised European and American Classification of Lymphoid Neoplasms' and the derived WHO classification are structured to mirror normal B/T-cell differentiation. In these modern classifications, distinct disease entities are defined based on the combination of morphology, immunological and molecular techniques and clinical features. The proposed major groups of lymphoid neoplasms are B-cell lymphomas/leukemias, T/Natural Killer-cell lymphomas/leukemias and Hodgkin disease. About 20 entities are recognised. This provides for the first time a truly international view of lymphomas and leukemias. It has emerged that such a classification can be used successfully by expert hematopathologists and yields highly reproducible results. It is also clear that no single marker, be it morphology, genetic analysis or immunophenotyping can be used as the 'gold standard' for diagnosis but that a combination of techniques is needed. PMID- 11397420 TI - Advances in the mass spectrometry of hemoglobin adducts: global analysis of the covalent binding of butadiene monoxide. AB - A common method to assess exposure to 1,3-butadiene through both occupational and environmental routes involves the detection of hemoglobin adducts formed by the primary reactive metabolite butadiene monoxide (EB). This assay is a modification of the Edman degradation procedure, which was developed to determine adducts formed specifically at the amine group of the N-terminal valine of hemoglobin. The goals of the current research are to determine the global modification of alpha- and beta-globin chains by EB and to localize the primary reactive residues to specific regions of the globin polypeptides. The degree of modification was monitored by electrospray mass spectrometry, which was used to measure the formation of EB-hemoglobin adducts (up to ten adducts per globin). Structural analysis of these modifications was performed by peptide mapping of globin peptides after trypsin digestion using liquid chromatography-mass spectrometry. These experiments provided information as to the relative reactivity of alpha- and beta-globin towards EB, as well as to the localization of adducts to specific peptide sequences. The results reveal variable reactivities of alpha- and beta globin towards EB and also show the formation of multiple adducts at several alpha- and beta-globin sites. In addition, it is established that the N-terminal valine residues are not the first to be modified by EB. PMID- 11397421 TI - Hemoglobin adducts as biomarkers of 1,3-butadiene in occupationally low exposed Italian workers and a few diesel-exposed miners. AB - Hemoglobin adducts were determined as biomarkers of 1,3-butadiene (BD) in 30 workers and 10 controls from an Italian BD plant and in 14 diesel-exposed miners. N-(2,3,4-trihydroxybutyl)valine (THBVal), an N-terminal valine globin adduct of reactive butadiene metabolites, was analyzed by gas chromatography/high resolution mass spectrometry after a modified Edman degradation and further acetylation. The BD exposure for the plant workers was 31 microg/m(3) (personal sampling). Whereas there was no detectable difference in hemoglobin adduct levels (range 17.7-61.4 pmol/g globin) between the total group of exposed and controls, slight but significant differences could be found between two subgroups of workers from different production units as well as one subgroup and controls (P<0.05), between smoking (n=13) and non-smoking exposed workers (n=17; P=0.066) as well as between smoking exposed workers and controls (P=0.055). Adduct levels of the miners (all non-smokers) were in the same range as those of the Italian BD workers and controls. The internal exposure and strain measured by THBVal levels resulting from a very low occupational BD exposure was in the range of the contribution of moderate smoking. PMID- 11397422 TI - Toxicokinetics of inhaled and endogenous isoprene in mice, rats, and humans. AB - Isoprene (IP) is ubiquitous in the environment and is used for the production of polymers. It is metabolized in vivo to reactive epoxides, which might cause the tumors observed in IP exposed rodents. Detailed knowledge of the body and tissue burden of inhaled IP and its intermediate epoxides can be gained using a physiological toxicokinetic (PT) model. For this purpose, a PT-model was developed for IP in mouse, rat, and human. Experimentally determined partition coefficients were taken from the literature. Metabolic parameters were obtained from gas-uptake experiments. The measured data could be described by introducing hepatic and extrahepatic metabolism into the model. At exposure concentrations up to 50 ppm, the rate of metabolism at steady-state is 14 times faster in mice and about 8 times faster in rats than in humans (2.5 micromol/h/kg at 50 ppm IP in air). IP does accumulate only barely due to its fast metabolism and its low thermodynamic partition coefficient whole body:air. IP is produced endogenously. This production is negligible in rodents compared to that in humans (0.34 micromol/h/kg). About 90% of IP produced endogenously in humans is metabolized and 10% is exhaled unchanged. The blood concentration of IP in non-exposed humans is predicted to be 9.5 nmol/l. The area under the blood concentration-time curve (AUC) following exposure over 8 h to 10 ppm IP is about 4 times higher than the AUC resulting from the unavoidable endogenous IP over 24 h. A comparison of such AUCs can be used for establishing workplace exposure limits. For estimation of the absolute risk, knowledge of the body burden of the epoxide intermediates of IP is required. Unfortunately, such data are not yet available. PMID- 11397423 TI - A novel DNA adduct, originating from 1,2-epoxy-3,4-butanediol, is the major DNA adduct after exposure to [2,3-(14)C]-1,3-butadiene,[4-(14)C]-1,2-epoxy-3-butane. AB - 1,3-Butadiene is a rodent carcinogen and its epoxide metabolites, 1,2-epoxy-3 butene (EB), 1,2-epoxy-3,4-butanediol (EBD), and 1,2:3,4-diepoxybutane (DEB) have been suggested as ultimate carcinogens. This study aimed at identification and quantification of DNA adducts in rats and mice following exposure to BD and its major metabolite EB to identify the reactive epoxide(s) in target tissues. Reaction of [4-(14)C]-EB with 2'-deoxyguanosine (dG) or DNA gave equal amounts of N7-(2-hydroxy-3-butenyl)guanine (G1) and N7-(1-(hydroxymethyl)-2-propenyl)guanine (G2). Reaction of DEB stereomers with dG yielded N7-(2,3,4 trihydroxybutyl)guanine (G3) as major adduct and novel, minor adduct (G4) that was tentatively identified as N7-(1-(hydroxymethyl)-2,3-dihydroxypropyl)guanine. For the stereomers of EBD, the opposite was found: reaction with dG led to G4 as major and G3 as minor adduct. 2D-Total correlation 1H-NMR spectroscopy of G4 indicated that the N7-alkyl group was in a virtually fixed conformational state and might interact with the O(6) of guanine, which might imply a higher degree of mutagenicity of G4, due to G-C mispairing, than of any other adducts observed in this study. At 48 h following exposure to [4-(14)C]-EB (1-50 mg/kg), DNA adduct profiles in rat and mouse were qualitatively similar, with G1 and G2 as main, and G4 as minor adduct. Following nose-only exposure to 200 ppm [2,3-(14)C]-BD for 6 h, G1 and G2 were minor adducts in liver (1.9 and 8.0 per 10(8) nucleotides) and lung (1.6 and 6.6 per 10(8) nucleotides, for rats and mice, respectively). G3 was absent in rats, but present in mouse liver and lung, at 20 and 12 adducts/10(8) nucleotides. The major adduct was G4 accounting for 13 and 90 (liver) and 11 and 139 (lung) adducts/10(8) nucleotides in rats and mice, respectively. Forty-two hours later, these adduct levels had only little changed. Our recent biomarker studies confirm that following exposure to BD, but not BDO, EBD is the major epoxide available for macromolecular binding in humans and rodents. Most probably because EBD is in contrast to EB and DEB, a poor substrate for epoxide hydrolases. In conclusion, the major DNA adduct following exposure to BD is G4, originating from EBD, and not from EB or BDE. It is concluded that EBD and G4 should be taken into account for human risk assessment for exposure to BD. PMID- 11397424 TI - Urinary metabolites and haemoglobin adducts as biomarkers of exposure to 1,3 butadiene: a basis for 1,3-butadiene cancer risk assessment. AB - Since 1,3-butadiene (BD) is a suspected human carcinogen, exposure to BD should be minimised and controlled. This study aimed at comparing the suitability of biomarkers for low levels of exposure to BD, and at exploration of the relative pathways of human metabolism of BD for comparison with experimental animals. Potentially sensitive biomarkers for BD are its urinary metabolites 1,2 dihydroxybutyl mercapturic acid (DHBMA, also referred to as MI) and 1- and 2 monohydroxy-3-butenyl mercapturic acid (MHBMA, also referred to as MII) and its haemoglobin (Hb) adducts 1- and 2-hydroxy-3-butenyl valine (MHBVal). In two field studies in BD-workers, airborne BD, MHBMA, DHBMA and MHBVal were determined. MHBMA proved more sensitive than DHBMA for monitoring recent exposures to BD and could measure 8-h time weighted average exposures as low as 0.13 ppm (0.29 mg/m(3)). The sensitivity of DHBMA was restricted by relatively high natural background levels in urine, of which the origin is currently unknown. MHBVal proved a sensitive method for monitoring cumulative exposures to BD at or above 0.35 ppm (0.77 mg/m(3)). Statistically significant relationships were found between either MHBMA or DHBMA and 8-h airborne BD levels, and between MHBVal adducts and average airborne BD levels over 60 days. The data showed a much higher rate of hydrolytic metabolism of BD in humans compared to animals, which was reflected in a much higher DHBMA/(MHBMA+DHBMA) ratio, and in much lower levels of MHBVal in humans, confirming in vitro results. Assuming a genotoxic mechanism, the data of this study coupled with our recent data on DNA and Hb binding in rodents, suggest that the cancer risk for humans from exposure to BD will be less than for the rat, and much less than for the mouse. PMID- 11397425 TI - In vitro genotoxicity testing of (1-chloroethenyl)oxirane, a metabolite of beta chloroprene. AB - (1-Chloroethenyl)oxirane (CEO) is a metabolite of beta-chloroprene (2-chloro-1,3 butadiene, CD). The purpose of this study was to evaluate the in vitro mutagenic and clastogenic (chromosome breaking) potential of CEO. For comparative purposes, the study also included an evaluation of the racemic compounds, 3,4-epoxy-1 butene (EB) and 1,2:3,4-diepoxybutane (DEB). Mutagenicity was evaluated in a bacterial reverse mutation test (Ames), using the pre-incubation method in the presence and absence of an exogenous metabolism system (Aroclor)-induced rat liver S9). Four Salmonella typhimurium tester strains, TA97a, TA98, TA100 and TA1535 were used. The exposure concentrations in the sealed incubation vials ranged from 0 to 69 mM for CEO, 0 to 102 mM for EB, and 0 to 83 mM for DEB. All three compounds showed signs of toxicity, with DEB being substantially more toxic than either CEO or EB. Mutagenic activity was observed with all three chemicals in primarily the base pair substitution strains (S. typhimurium TA100 and TA1535), but some activity was also seen in the frameshift elimination strains (S. typhimurium TA97a and TA98). The observed mutagenic responses after exposure with CEO or EB were greater than the observed response for DEB, most likely because of the higher toxicity of DEB. Generally, the mutagenic responses were unchanged in the frameshift strains and base pair substitution strains in the presence of S9 metabolism. In vitro clastogenicity was evaluated using the cytochalasin-B blocked micronucleus test in cultured Chinese hamster V79 cells. The test was conducted without S9 metabolism because of the absence of substantial changes in the Ames test. Exposure concentrations ranged from 0 to 0.943 mM for CEO, 0 to 3.0 mM for EB, and 0 to 0.035 mM for DEB, with the upper exposure concentrations dictated by cytotoxicity. Cytotoxicity, measured as a reduction in the proportion of binucleated cells and altered cell morphology, was observed for CEO at concentrations > or =0.175 mM. Exposure to EB led to a reduced proportion of binucleated cells at concentrations > or =2.0 mM, and cell death was observed after DEB exposure at concentrations > or =0.025 mM. No clastogenicity was observed in the V79 cells when tested up to cytotoxic concentrations of CEO, whereas an elevated frequency of micronuclei was observed after exposure to either EB (> or =1.0 mM) or DEB (> or =0.0125 mM). These results suggest that CEO-induced mutagenicity, but not clastogenicity, may contribute to the observed beta-chloroprene-induced carcinogenicity in the rodent bioassay studies. PMID- 11397426 TI - Global phytochemistry: the Brazilian approach. PMID- 11397427 TI - Primary sequence determination of a Kunitz inhibitor isolated from Delonix regia seeds. AB - A serine proteinase inhibitor was purified from Delonix regia seeds a Leguminosae tree of the Caesalpinioideae subfamily. The inhibitor named DrTI, inactivated trypsin and human plasma kallikrein with K(i )values 2.19x10(-8) M and 5.25 nM, respectively. Its analysis by SDS-PAGE 10-20% showed that the inhibitor is a protein with a single polypeptide chain of M(r) 22 h Da. The primary sequence of the inhibitor was determined by Edman degradation, thus indicating that it contained 185 amino acids and showed that it belongs to the Kunitz type family; however, its reactive site did not contain Arg or Lys at the putative reactive site (position 63, SbTI numbering) or it was displaced when compared to other Kunitz-type inhibitors. PMID- 11397428 TI - Vanadium haloperoxidases from brown algae of the Laminariaceae family. AB - Vanadium haloperoxidases were extracted, purified and characterized from three different species of Laminariaceae--Laminaria saccharina (Linne) Lamouroux, Laminaria hyperborea (Gunner) Foslie and Laminaria ochroleuca de la Pylaie. Two different forms of the vanadium haloperoxidases were purified from L. saccharina and L. hyperborea and one form from L. ochroleuca species. Reconstitution experiments in the presence of several metal ions showed that only vanadium(V) completely restored the enzymes activity. The stability of some enzymes in mixtures of buffer solution and several organic solvents such as acetone, ethanol, methanol and 1-propanol was noteworthy; for instance, after 30 days at least 40% of the initial activity for some isoforms remained in mixtures of 3:1 buffer solution/organic solvent. The enzymes were also moderately thermostable, keeping full activity up to 40 degrees C. Some preliminary steady-state kinetic studies were performed and apparent Michaelis-Menten kinetic parameters were determined for the substrates iodide and hydrogen peroxide. Histochemical studies were also performed in fresh tissue sections from stipe and blade of L. hyperborea and L. saccharina, showing that haloperoxidase activity was concentrated in the external cortex near the cuticle, although some activity was also observed in the inner cortical region. PMID- 11397429 TI - Endoplasmic oleoyl-PC desaturase references the second double bond. AB - The regiospecificity for the gene product of fad2,(1) the microsomal oleoyl-PC desaturase from higher plants, differs from some previous suggestions. Rather than only referencing the carboxyl group (a Delta(12) desaturase) or the methyl terminus (an omega-6 desaturase), this desaturase locates the second double bond in its substrates by first referencing the existing double bond. This specificity was demonstrated for the oleoyl-PC desaturase cDNA from the developing seeds of peanut (Arachis hypogaea L) expressed in yeast (Saccharomyces cerevisae). The expressed enzyme was capable of desaturating monounsaturated fatty acyl groups in membrane lipids. Endogenous palmitoleate was desaturated to cis, cis 9,12 hexadecadienoate (9(Z)12(Z)C16:2), endogenous oleate to linoleate (9(Z)12(Z) octadecadienoate), and cis 10-nonadecenoate (provided as a supplement in the growth medium) to 10(Z)13(Z)C19:2. The rule, Delta(x+3) where x=9 is the double bond location in the substrate, best describes the consistent placement of the second double bond in the above monounsaturated substrates for the oleoyl-PC desaturase of higher plants. PMID- 11397430 TI - Main glucosidase conversion products of the gluco-alkaloids dolichantoside and palicoside. AB - The enzymatic glucose cleavage of palicoside revealed the biosynthetic pathway to akagerine, whereas the conversion of dolichantoside led to a new quaternary heteroyohimbine alkaloid named N(b)-methyl-21-beta-hydroxy-mayumbine. The hypothetical models of reactions occurring after the conversion of both substrates are proposed. Dolichantoside and palicoside, as well as Strychnos mellodora stem bark crude ethanol extract, exhibit significant antimycotic activity against human pathogens in presence of specific glucosidase. PMID- 11397431 TI - Structure of fructans from excised leaves of New Zealand flax. AB - The accumulation of total water-soluble carbohydrate, and specifically sucrose and fructan, by excised leaves of Phormium tenax and P. cookianum (family Phormiaceae J. G. Agardh, order Asparagales) was investigated. Total water soluble carbohydrate content of excised leaves of P. tenax and P. cookianum increased during 48 h of continuous illumination at an average rate of 1.3 and 0.9 mg g(-1) fresh weight leaf per hour, respectively. The sucrose content of excised leaves increased throughout the experimental period. The fructan content of excised leaves of P. tenax increased slightly throughout the experimental period, whilst that of P. cookianum was variable and showed no overall change. Chemical and spectroscopic analysis of the fructans obtained from the two Phormium species showed that they were similar to each other and contained mostly 1-linked and terminal fructofuranosyl (Fruf) residues, together with smaller amounts of 6-linked Fruf, 1,6-branched Fruf, terminal and 6-linked glucopyranosyl residues. Separation of the fructans by thin-layer and high-performance anion exchange chromatography revealed the presence of a complex mixture of fructo oligosaccharides and higher molecular weight fructan. The branched structure of the fructans isolated from excised leaves of Phormium resembles that of fructans and fructo-oligosaccharides isolated from some related species within the order Asparagales (Agave vera cruz, Cordyline australis and Urginea maritima), but is distinct from the linear structure of fructans from others (Allium cepa and Asparagus officinalis). The structural heterogeniety of fructans within both the order Asparagales and superorder Liliiflorae may be a useful chemotaxonomic aid. PMID- 11397432 TI - Biotransformation of (+)- and (-)-camphorquinones by plant cultured cells. AB - Biotransformation of (+)- and (-)-camphorquinones with suspension plant cultured cells of Nicotiana tabacum and Catharanthus roseus was investigated. It was found that the plant cultured cells of N. tabacum and C. roseus reduce stereoselectively the carbonyl group of (+)- and (-)-camphorquinones to the corresponding alpha-keto alcohols. PMID- 11397433 TI - Biotransformation of geranyl acetate to geraniol during palmarosa (Cymbopogon martinii, Roxb. wats. var. motia) inflorescence development. AB - Only immature palmarosa (Cymbopogon martinii, Roxb. wats. var. motia) inflorescence with unopened spikelets accumulated essential oil substantially. Geraniol and geranyl acetate together constituted about 90% of the palmarosa oil. The proportion of geranyl acetate in the oil decreased significantly with a corresponding increase of geraniol, during inflorescence development. An esterase enzyme activity, involved in the transformation of geranyl acetate to geraniol, was detected from the immature inflorescence using a gas chromatographic procedure. The enzyme, termed as geranyl acetate cleaving esterase (GAE), was found to be active in the alkaline pH range with the optimum at pH 8.5. The catalysis of geranyl acetate was linear up to 6 h, and after 24 h of incubation, 75% of the geranyl acetate incubated was hydrolyzed. The GAE enzymic preparation, when stored at 4 degrees C for a week, was quite stable with only 40% loss of activity. The physiological role of GAE in the production of geraniol during palmarosa inflorescence development has been discussed. PMID- 11397434 TI - Seasonal variation of monoterpene emission from Malus domestica and Prunus avium. AB - Emission rates of monoterpenes released by apple (Malus domestica Borkh) and cherry (Prunus avium L.) were estimated at different phenological stages. These measurements employed a dynamic flow-through Teflon chamber, sample collection onto cartridges filled with graphitized carbon and thermal desorption gas chromatography-mass spectrometry (GC-MS) for identification and quantification of the emitted volatiles. At full bloom the release of monoterpene hydrocarbons from cherry flowers was 1213 ng g(-1) dry weight (DW) h(-1), exceeding by approximately three-fold the emission rate of apple flowers (366 ng g(-1) DW h( 1)). Observed seasonal variations in biogenic volatile organic compound (VOC) emissions ranged over several order of magnitudes. At fruit-set and ripening stages, in fact, the hydrocarbon emission dramatically decreased reaching the lowest values at harvest time when leaves were fully mature (3-9 ng g(-1) DW h( 1)). Wide diversity in the composition of compounds from the species studied was also recorded. At blooming, linalool contributed significantly to the monoterpene emission from apple (94% of the emitted carbon) while alpha-pinene and camphene represented on average more than 60% of the total emitted volatiles from cherry flowers. Among the monoterpenes identified in flowers, alpha-pinene, camphene and limonene were also found in the foliage emission of both species. Fruit trees are relevant monoterpene emitters only at blooming and thus for a short period of the vegetative cycle. When leaves are fully developed, the carbon loss due to monoterpene emissions related to the photosynthetically carbon gain is negligible. PMID- 11397435 TI - Botrydial is produced in plant tissues infected by Botrytis cinerea. AB - The fungal metabolite botrydial was detected for the first time in ripe fruits of sweet pepper (Capsicum annuum) wound-inoculated with conidial suspensions of Botrytis cinerea and also in leaves of Phaseolus vulgaris and Arabidopsis thaliana inoculated without wounding. This phytotoxin was produced in soft rot regions of the infection. In C. annuum, the most aggressive isolate produced the highest botrydial concentrations in planta. The levels of botrydial produced by this isolate did not correlate with the reported relative susceptibilities of four P. vulgaris genotypes. The results suggest that botrydial is a pathogenicity factor for this fungus, but not a primary determinant of pathogenicity. PMID- 11397436 TI - Identification of glucosinolates on the leaf surface of plants from the Cruciferae and other closely related species. AB - Leaf-surface extracts prepared from 18 non-cultivated (wild) plant species, derived from the Capparidaceae, Cruciferae, Resedaceae and Tropaeolaceae were ranked for their ability to stimulate oviposition by the cabbage root fly, and analysed for glucosinolates. A total of 28 different glucosinolates were identified. A clear relationship was detected between the indolyl-, benzyl- and the total glucosinolate composition on the leaf surface and oviposition preference by cabbage root fly females. However, as the results are not fully explained by differences in leaf surface glucosinolates, other important oviposition deterrents and stimuli on the leaf surface of these wild crucifers must also be present. PMID- 11397437 TI - Antioxidant enzymes responses to cadmium in radish tissues. AB - To investigate the antioxidant responses of radish (Raphanus sativus L.) to cadmium (Cd) treatment, seedlings of a tolerant variety were grown in increasing concentrations of CdCl(2), ranging from 0.25-1 mM, for up to 72 h in a hydroponic system. Analysis of Cd uptake indicated that most of the Cd accumulated in the roots, but some was also translocated and accumulated in the leaves, especially at the higher concentrations of Cd used in the experiments. Roots and leaves were analysed for catalase, glutathione reductase and superoxide dismutase activities. Catalase and glutathione reductase activities increased considerably in the roots and leaves after 24 h exposure to the metal, indicating a direct correlation with Cd accumulation. The analysis of native PAGE enzyme activity staining, revealed several superoxide dismutase isoenzymes in leaves, with the two predominant isoenzymes exhibiting increases in activity in response to Cd treatment. The results suggest that in radish, the activity of antioxidant enzymes responds to Cd treatment. The main response may be via the activation of the ascorbate glutathione cycle for the removal of hydrogen peroxide, or to ensure the availability of glutathione for the synthesis of Cd-binding proteins. PMID- 11397438 TI - 3-O-methyl-D-galactose residues in lycophyte primary cell walls. AB - Acid hydrolysis of cell wall-rich material from young leaves of the lycophyte Selaginella apoda (L.) Spring yielded substantial amounts of 3-O-methyl-D galactose (1) in addition to the usual major monosaccharides (glucose, galactose, arabinose, xylose and galacturonic acid). The yield of 1 approximately equalled that of galacturonic acid. Compound 1 was identified as 3-O-methylgalactose by its 1H and 13C NMR spectra, and shown to be the D-enantiomer by its susceptibility to D-galactose oxidase. Compound 1 was detected in acid hydrolysates of the alcohol-insoluble residues from young leaves of all lycophytes tested, both homosporous (Lycopodium, Huperzia and Diphasiastrum) and heterosporous (Selaginella). It was not detectable in the charophyte green algae Coleochaete scutata, Chara coralina or Klebsormidium flaccidum, any bryophytes [a hornwort (Anthoceros), four liverworts and three mosses], or any euphyllophytes [a psilopsid (Psilotum), a horsetail (Equisetum), eusporangiate and leptosporangiate ferns, the gymnosperm Gnetum, and diverse angiosperms]. A high content of 1 is thus an autapomorphy of the lycophytes. PMID- 11397439 TI - Dihydroxynortropane alkaloids from calystegine-producing plants. AB - Three dihydroxynortropanes, 2alpha,7beta-dihydroxynortropane, 2alpha,3beta dihydroxynortropane, and 3alpha,7beta-dihydroxynortropane, were isolated from calystegine-producing plants in the families Convolvulaceae and Solanaceae. 2alpha,7beta-Dihydroxynortropane was isolated from six species in the Convolvulaceae whereas only Calystegia soldanella contained it and 2alpha,3beta dihydroxynortropane. Although neither of these were detectable in three species tested in the Solanaceae, 3alpha,7beta-dihydroxynortropane was, however, isolated from Duboisia leichhardtii. PMID- 11397440 TI - Fatty acid profiles and their chemotaxonomy in planktonic species of Anabaena (Cyanobacteria) with straight trichomes. AB - Twenty-four axenic strains of planktonic Anabaena with straight trichomes, assigned to 7 species, were investigated by analyzing the pattern and content of their fatty acid composition and comparing their fatty acid composition with their morphological properties. In general, the fatty acids in planktonic Anabaena contained 14:0, 16:0, 16:1(cis-), 18:0, 18:1, 18:2, and 18:3(alpha) as their major components, and were classified as Type 2 according to the Kenyon Murata system. These strains were further divided into 2 subtypes: 18 strains belonging to Type 2A, which contains 16:2 and 16:3, and 6 strains to Type 2B, which lacks 16:2 and 16:3. Fatty acid compositions of strains of A. solitaria, A. smithii, and A. kisseleviana closely corresponded to morphological properties; however, 10 strains of A. planctonica were divided into 4 clusters, and 3 strains of A. affinis into 2 clusters. These clusters should be taxonomically evaluated based on other aspects such as genetic characteristics. PMID- 11397441 TI - Chemical variability in the essential oil of Hyptis suaveolens. AB - The essential oils of Hyptis suaveolens plants collected from 11 localities of the Brazilian Cerrado region were investigated by GC-MS. Sabinene, limonene, biclyclogermacrene, beta-phellandrene and 1,8-cineole were the principal constituents. The results were submitted to principal component and chemometric cluster analysis which allowed three groups of essential oils to be distinguished with respect to the content of p-mentha-2,4(8)-diene, limonene/beta phellandrene/gamma-terpinene and germacrene D/bicyclogermacrene. In patterns of geographic variation in essential oil composition indicated that the sesquiterpenes are mainly produced in the samples grown at lower latitudes. PMID- 11397442 TI - 3',5'-Di-C-beta-glucopyranosylphloretin, a flavonoid characteristic of the genus Fortunella. AB - Dihydrochalcone derivative, 3',5'-di-C-beta-glucopyranosylphloretin (1), is present in the genus Fortunella, (F. crassifolia, F. japonica, F. margarita, F. polyandra and F. hindsii). These species accumulate a large quantity of 1 in their fruits (peel, 6.5-15.2 mg/g in dry wt; juice sac, 1.5-10.5 mg/g) and in their leaves (21.3-60.2 mg/g). Twenty-seven Tanaka's Citrus species examined lack 1, but C. madurensis and C. halimii contain 1 in large quantities in their peels (25.1 and 33.6 mg/g) and juice sacs (4.1 and 4.2 mg/g). Poncirus species do not contain 1. Fortunella-citrus hybrids, the Orangequat [C. unshiuxF. crassifolia], the Thomasville citrangequat [Fortunella sp.x(C. sinensisxPoncirus trifoliata)], and seven hybrid progenies [F. margaritaxC. junos], contain large amounts of 1 in their peels (17.0-7.9 mg/g) and juice sacs (2.0-9.9 mg/g). These facts suggest that accumulation of 1 is a generic trait of the genus Fortunella and that the inheritance of the trait among the intergeneric hybrids is controlled by a dominant allele. Thus C. madurensis and C. halimii are thought to originate from natural hybrids between the genera Citrus and Fortunella. Phloridzin, which has the same aglycon as 1, was not detected in the citrus plants examined. PMID- 11397443 TI - Antifungal polysulphides from Petiveria alliacea L. AB - Bioactivity-directed fractionation of the CH(2)Cl(2)/MeOH (2:1, v/v) extract of the roots of Petiveria alliacea, using mutant yeast strains of Saccharomyces cerevisiae and fungi Cladosporium cladosporioides and C. sphaerospermum led to the isolation of dipropyl disulphide (1), dibenzyl sulphide (2), dibenzyl disulphide (3), dibenzyl trisulphide (4), dibenzyl tetrasulphide (5), benzylhydroxymethyl sulphide (6) and di(benzyltrithio) methane (7). Of these, 5-7 are new compounds and this is the first report of the natural occurrence of 2 and 3. PMID- 11397444 TI - Endogenous gibberellins in immature seeds of Prunus persica L.: identification of GA(118), GA(119), GA(120), GA(121), GA(122) and GA(126). AB - The endogenous gibberellins in immature seeds of Prunus persica were analyzed by gas chromatography-mass spectrometry. Eleven known gibberellins, GA(3), GA(9), GA(17), GA(19), GA(30), GA(44), GA(61), GA(63), GA(87), GA(95) and GA(97) were identified. Additionally, several hitherto unknown gibberellins were detected and their putative structures were verified by synthesis of the authentic gibberellins. These gibberellins were then assigned trivial numbers, e.g. 1alpha hydroxy GA(20) (GA(118)), 1alpha-hydroxy GA(9) (GA(119)), 1,2-didehydro GA(9) (GA(120)), 1,2-didehydro GA(70) (GA(121)), 1,2-didehydro GA(69) (GA(122)) and 1,2 didehydro GA(77) (GA(126)). GA(118) and GA(119) were the first 1alpha-hydroxy gibberellins identified from higher plants. The above profile of 1,2-didehydro gibberellins suggests that 1,2-dehydrogenation might occur prior to 3beta hydroxylation in biosynthesis of GA(3), GA(30) and GA(87) in immature seeds of P. persica. PMID- 11397445 TI - Occurrence of cadaverine in hairy roots of Brugmansia candida. AB - The polyamine, cadaverine, was detected in transformed root cultures of Brugmansia candida (syn. Datura candida), a Solanaceae which produces the tropane alkaloids scopolamine and hyoscyamine. To the best of our knowledge, this is the first time that the existence of this uncommon polyamine has been detected in a Datura species. Cadaverine, however, could not be found in the whole plant. The occurrence of cadaverine in hairy roots could be a consequence of either the transformation or a response to stress. Also, cadaverine could be participating in other secondary pathways rather than to the tropane alkaloids. The common polyamines, putrescine, spermidine and spermine were also observed. PMID- 11397446 TI - The phenolic fraction of maize bran: evidence for lignin-heteroxylan association. AB - Maize bran heteroxylan samples were extracted in various conditions of severity. Their ferulate and diferulate content was investigated by GC-MS of methyl ester TMSi derivatives. When extracted by 0.5 M NaOH in mild conditions, the heteroxylan sample contained a low level of ferulic acid (0.032% by wt.) and the main diferulate surviving alkaline extraction was found to be the 8-8' diferulate. On peroxidase treatment, this sample nevertheless produced a firm and brittle gel without any change in the diferulate profile. Typical lignin structures, mainly comprising syringyl units interconnected through beta-O-4, beta-1 and beta-beta interunit bonds, were evidenced in the maize bran sample. More importantly, these lignin structures were found to be tightly associated with the alkali-extracted heteroxylans. Thioacidolysis revealed the occurrence of 0.1-0.5% (by wt.) lignin structures in heteroxylan fractions extracted in mild or severe conditions, before and after purification of the polysaccharides. The gelling potential of the heteroxylan fractions was not only dependent on their ferulate level, but also influenced by associated lignin structures. These results argue for the occurrence of covalent linkages between heteroxylan chains and lignin structures which could participate in the peroxidase-driven gelation of feruloylated polysaccharides. They demonstrate the role of low lignin levels in the organization of native or reconstructed polysaccharide networks. PMID- 11397448 TI - Investigation of Una De Gato I. 7-Deoxyloganic acid and 15N NMR spectroscopic studies on pentacyclic oxindole alkaloids from Uncaria tomentosa. AB - The C-8-(S) isomer of deoxyloganic acid (7-deoxyloganic acid), together with beta sitosteryl glucoside, five known stereoisomeric pentacyclic oxindole alkaloids and the tetracyclic oxindole isorhyncophylline, were isolated from the inner bark of Uncaria tomentosa. Structures of the isolated compounds were based on 1H and 13C NMR data, mainly 2D NMR experiments, including 1H-13C HMBC and 1H-1H NOESY correlation. Furthermore, the hitherto unreported 15N chemical shifts of the isomeric oxindole alkaloids, using 1H-15N HMBC experiments, were utilized to facilitate their characterization. Uncarine D showed weak cytotoxic activity against SK-MEL, KB, BT-549 and SK-OV-3 cell lines with IC(50) values between 30 and 40 microg/ml, while uncarine C exhibited weak cytotoxicity only against ovarian carcinoma (IC(50) at 37 microg/ml). PMID- 11397447 TI - Triterpene glycosides from the roots of Sanguisorba officinalis. AB - Five triterpene glycosides were isolated from the MeOH extract of Sanguisorba officinalis (Rosaceae) roots, as confirmed by detailed analysis of their 1H, 13C, and two-dimensional NMR data, and by the results of hydrolytic cleavage. Three known triterpenes and six known triterpene glycosides were also isolated and identified. The isolated compounds were evaluated for their cytotoxic activities against HSC-2 cells and HGF. PMID- 11397449 TI - Sterol composition of Aureoumbra lagunensis, the Texas brown tide alga. AB - Aureoumbra lagunensis, the alga responsible for the "Texas brown tide", contains (E)-24-propylidenecholesterol (35.7% of total sterols) as its dominant sterol, in common with other members of the Pelagophyceae. Other major sterols are stigmasterol (22.2%), sitosterol (19.2%), cholesterol (14.1%), and (24R)-24 propylcholesterol (5.2%). Trace amounts of 24-methylenecholesterol, crinosterol, clerosterol, campesterol, dihydrobrassicasterol, and 24-isopropylcholesterol were also detected. PMID- 11397450 TI - Influence of trans-cis isomerisation of coumaric acid substituents on colour variance and stabilisation in anthocyanins. AB - The recently isolated pigments from Petunia integrifolia and Triteleia bridgesii present a distinct feature that sheds new light on the understanding of intramolecular copigmentation of anthocyanins. These are among the infrequent anthocyanins that naturally present a coumaric acid substituent in both cis and trans forms. As a consequence, the two isomers demonstrate substantial variations of their thermodynamic and kinetic constants and also colour properties. A possible explanation for these characteristics is presented, making use of molecular modelling and taking into account the three-dimensional structures of the pigments. PMID- 11397454 TI - Humicola insolens cellobiose dehydrogenase: cloning, redox chemistry, and "logic gate"-like dual functionality. AB - 1Cellobiose dehydrogenase is a hemoflavoenzyme that catalyzes the sequential electron-transfer from an electron-donating substrate (e.g. cellobiose) to a flavin center, then to an electron-accepting substrate (e.g. quinone) either directly or via a heme center after an internal electron-transfer from the flavin to heme. We cloned the dehydrogenase from Humicola insolens, which encodes a protein of 761 amino acid residues containing an N-terminal heme domain and a C terminal flavin domain, and studied how the catalyzed electron transfers are regulated. Based on the correlation between the rate and redox potential, we demonstrated that with a reduced flavin center, the enzyme, as a reductase, could export electron from its heme center by a "outer-sphere" mechanism. With the "resting" flavin center, however, the enzyme could have a peroxidase-like function and import electron to its heme center after a peroxidative activation. The dual functionality of its heme center makes the enzyme a molecular "logic gate", in which the electron flow through the heme center can be switched in direction by the redox state of the coupled flavin center. PMID- 11397453 TI - Purification and properties of a novel raw starch degrading-cyclodextrin glycosyltransferase from Klebsiella pneumoniae AS- 22. AB - A novel raw starch degrading alpha-cyclodextrin glycosyltransferase (CGTase; E.C. 2.4.1.19), produced by Klebsiella pneumoniae AS-22, was purified to homogeneity by ultrafiltration, affinity and gel filtration chromatography. The specific cyclization activity of the pure enzyme preparation was 523 U/mg of protein. No hydrolysis activity was detected when soluble starch was used as the substrate. The molecular weight of the pure protein was estimated to be 75 kDa with SDS-PAGE and gel filtration. The isoelectric point of the pure enzyme was 7.3. The enzyme was most active in the pH range 5.5-9.0 whereas it was most stable in the pH range 6-9. The CGTase was most active in the temperature range 35-50 degrees C. This CGTase is inherently temperature labile and rapidly loses activity above 30 degrees C. However, presence of soluble starch and calcium chloride improved the temperature stability of the enzyme up to 40 degrees C. In presence of 30% (v/v) glycerol, this enzyme was almost 100% stable at 30 degrees C for a month. The K(m) and k(cat) values for the pure enzyme were 1.35 mg ml(-1) and 249 &mgr;M mg( 1) min(-1), respectively, with soluble starch as the substrate. The enzyme predominantly produced alpha-cyclodextrin without addition of any complexing agents. The conditions employed for maximum alpha-cyclodextrin production were 100 g l(-1) gelatinized soluble starch or 125 g l(-1) raw wheat starch at an enzyme concentration of 10 U g(-1) of starch. The alpha:beta:gamma-cyclodextrins were produced in the ratios of 81:12:7 and 89:9:2 from gelatinized soluble starch and raw wheat starch, respectively. PMID- 11397455 TI - Effect of gluconic acid as a secondary carbon source on non-growing L-lysine producers cells of Corynebacterium glutamicum. Purification and properties of 6 phosphogluconate dehydrogenase. AB - We studied the production of L-lysine in Corynebacterium glutamicum ATCC 21543 non growing cells obtained by nutrient limitation. Statistical analysis revealed significant differences in the L-lysine titers of glucose, gluconic acid or glucose-gluconic acid cultures. Higher L-lysine titer obtained in batch cultures with mixed carbon sources or gluconic acid alone were found to be associated with a high 6-phosphogluconate dehydrogenase activity (6PGDH, E.C.1.1.1.44). This enzyme is a pivotal enzyme within the hexose monophosphate pathway, and thus of importance for L-lysine production. 6PGDH was purified and characterized. The purified enzyme migrates as a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis with a molecular mass of 52.5 kDa. The molecular mass of the native enzyme was estimated to be 120 kDa by molecular exclusion chromatography, thus suggesting a homodimeric structure. The amino terminal sequence shows a strong similarity (a match of 86% of the first 20 amino acid) to the 6PGDH from other microorganisms such as, E. coli and B. subtilis. The pI of the dimeric native enzyme and the optimum pH were 6.2 and 8.0, respectively. For the oxidative decarboxylation of 6-phosphogluconate, K(m) of 71 &mgr;M and 43 &mgr;M were obtained for 6-phosphogluconate and NADP(+), respectively. PMID- 11397456 TI - Wet strength improvement of unbleached kraft pulp through laccase catalyzed oxidation. AB - Previous investigations have shown that laccase catalyzed oxidation of lignin containing wood fibers can enhance the strength of medium density fiberboards. In the present work it was investigated if laccase treatment had any impact on the tensile strength of a high yield unbleached kraft pulp. Treatment with laccase alone had only a very little effect on the wet strength of the pulp, whereas addition of lignin rich extractives increased the wet strength after the enzyme treatment significantly. A mediated oxidation gave a similar improvement of the wet tensile strength although no lignin was added to the fiber suspension. Furthermore, it was found that a heat treatment combined with a mediated oxidation gave a higher improvement in wet tensile strength than could be accounted for by the individual treatments. No change in dry tensile strength from the laccase treatment was observed. It is suggested that the observed improvement in wet tensile strength is related to polymerization of lignin on fibers in the hand sheet and/or coupling of phenoxy radicals on lignin associated to adjacent fibers. For the different mediators studied, a correlation was found between oxygen consumption upon mediated oxidation and generation of wet strength in the pulp. PMID- 11397457 TI - Phage display combinatorial libraries of short peptides: ligand selection for protein purification. AB - A library of heptapeptides displayed on the surface of filamentous phage M13 was evaluated as a potential source of affinity ligands for the purification of Rhizomucor miehei lipase. Two independent selection (biopanning) protocols were employed: the enzyme was either physically adsorbed on polystyrene or chemically immobilized on small magnetic beads. From screening with the polystyrene-adsorbed lipase it was found that there was a rapid enrichment of the library with "doublet" clones i.e. the phage species which carried two consecutive sequences of heptapeptides, whilst no such clones were observed from the screening using lipase attached to magnetic beads. The binding of the best clones to the enzyme was unambiguously confirmed by ELISA. However the synthetic heptapeptide of identical sequence to the best "monomeric" clone did not act as a satisfactory affinity ligand after immobilization on Sepharose. This indicated that the interaction with lipase was due to both the heptapeptide and the presence of a part of the phage coat protein. This conclusion was further verified by immobilizing the whole phage on the surface of magnetic beads and using the resulting conjugate as an affinity adsorbent. The scope of application of this methodology and the possibility of preparing phage-based affinity materials are briefly discussed. PMID- 11397458 TI - Isothermal and non-isothermal characterization of catalytic nylon membranes chemically grafted: dependence on the grafting percentage. AB - The behaviour of five different hydrophobic beta-galactosidase derivatives, obtained by grafting different amount of butylmethacrylate (BMA) on planar nylon membranes, has been studied under isothermal and non-isothermal conditions.Under isothermal conditions the effect of the grafting percentage on the enzyme activity has been studied as a function of pH, temperature and substrate concentration. Independently from the parameters under observation, the yield of the catalytic process reaches the maximum value at a grafting percentage value equal to 21%. The apparent K(m) values result linearly increasing with the increase of the grafting percentage, while the apparent V(max) exhibits a maximum value.Under non-isothermal conditions, a decrease of the apparent K(m) values and increase of the apparent V(max) has been found in respect to the same values obtained under isothermal conditions.The percentage activity increases induced by the presence of a temperature gradient have been found to decrease with the increase of the percentage of graft BMA.A parameter correlating the percentage increase of enzyme activity under non-isothermal conditions with the hydrophobicity of the catalytic membrane has also been identified. This parameter is the ratio between thermoosmotic and hydraulic permeability.Results have been discussed in terms of reduction of diffusion limitations for substrate and products movement towards or away from the catalytic site by the process of thermodialysis.The usefulness of using non-isothermal bioreactors in industrial biotechnological processes has been confirmed. PMID- 11397460 TI - An efficient purification process for sweet potato beta-amylase by affinity precipitation with alginate. AB - beta-amylases are used in production of maltose syrup. It is shown that sweet potato beta-amylase can be purified by affinity precipitation with alginate with 80% activity yield and 44 fold purification. SDS-PAGE of the purified protein showed a single band and a subunit weight of 50 kDa. Preliminary data with soybean and barley enzymes indicate that this may be a general method for purification of beta-amylases. PMID- 11397459 TI - L(-)-carnitine production using a recombinant Escherichia coli strain. AB - The L(-)-carnitine production by biotransformation using the recombinant strain Escherichia coli pT7-5KE32 has been studied and optimized with crotonobetaine and D(+)-carnitine as substrates. A resting rather than a growing cells system for L( )-carnitine production was chosen, crotonobetaine being the best substrate. High biocatalytic activity was obtained after growing the cells under anaerobic conditions at 37 degrees C and with crotonobetaine or L(-)-carnitine as inducer. The growth incubation temperature (37 degrees C) was high enough as to activate the heat-inducible lambdap(L) promoter inserted in the plasmid pGP1-2. The best biotransformation conditions were with resting cells, under aerobiosis, with 4 g l(-1) and 100 mM biomass and substrate concentrations respectively. Under these conditions the biotransformation time (1 h) was shorter and the L(-)-carnitine yield (70%) higher than previously reported. Consequently productivity value (11.3 g l(-1)h(-1)) was highly improved when comparing with other published works. The resting cells could be reused until eight times maintaining product yield levels well over 50% that meant to increase ten times the L(-)-carnitine obtained per gram of biomass. PMID- 11397461 TI - The application of continuous culture for plant cell suspensions. AB - Continuous culture of plant cell suspensions has been developed during the last 35 years. Starting from rather imperfect set-ups, nowadays much better equipment is used for studies on growth and production kinetics or cell physiology. In this review the development of equipment and theory, as well as the applications are discussed. PMID- 11397462 TI - Effect of cell membrane of Agrobacterium radiobacter on enhancing D-amino acids production from racemic hydantoins. AB - An interesting phenomenon was observed that the existence of the intact cell membrane can enhance the D-amino acids production from D,L-5-substituted hydantoins by reacting with the whole cells of Agrobacterium radiobacter. Two intracellular enzymes were involved in the reaction process. The first enzyme D hydantoinase converted hydantoins to carbamoyl derivatives which were further converted to D-amino acids by D-amidohydrolase. The amount of D-amino acids produced from hydantoins by the intact cells were 1.8-2.4 fold higher than the toluene treated cells. In addition, by using the intact cells the amount of D amino acids produced from hydantoins was about 10 fold higher than that produced directly from carbamoyl derivatives. The relatively lower permeability of cell membrane to the reaction intermediate carbamoyl derivatives was confirmed by a simple mathematical model to be the main factor for the better performance of the intact cells for D-amino acid production. Besides, the low intracellular enzymes activities also contributed to the effect of intact cell membrane on enhancing the D-amino acid production. PMID- 11397463 TI - Immobilization of catalases from Bacillus SF on alumina for the treatment of textile bleaching effluents. AB - A catalase preparation from a newly isolated Bacillus sp. was covalently immobilized on silanized alumina using glutaraldehyde as crosslinking agent. The effect of the coupling time of the enzyme-support reaction was determined in terms of protein recovery and immobilization yield and a certain balance point was found after which the activity recovery decreased. The activity profile of the immobilized catalase at high pH and temperature was investigated. The immobilized enzyme showed higher stabilities (214 h at pH 11, 30 degrees C) at alkaline pH than the free enzyme (10 h at pH 11, 30 degrees C). The immobilized catalase was inhibited by anionic stabilizers or surfactants added to the hydrogen peroxide substrate solution. PMID- 11397464 TI - Lysozyme inactivation under mechanical stirring: effect of physical and molecular interfaces. AB - This article focuses on the role of interfaces on lysozyme inactivation and aggregation process in stirred reactor. The first order inactivation constant of this process has found to be proportional not only to the power imparted by the impeller but also to the area of glass-liquid, air-liquid and PTFE-liquid interfaces in three reactors. Both area and type of interfaces act on inactivation: PTFE and air are four more efficient than glass to promote lysozyme inactivation because of their hydrophobicity. As well as physical interfaces, molecular surfaces of inactivated enzymes -more hydrophobic than native enzymes- enhance lysozyme inactivation and aggregation. This enhancement has been found to be correlated with the properties of aggregates of inactivated enzymes, especially their number. Then, under mechanical stirring, inactivation aggregation process is induced by physical interfaces and self-catalyzed by increasing hydrophobic surfaces of inactivated enzymes. PMID- 11397465 TI - Batch cultures of supplemented whey permeate using Lactobacillus helveticus: unstructured model for biomass formation, substrate consumption and lactic acid production. AB - The Luedeking and Piret expression can not account for the cessation of production observed at the end of batch; so an empiric term has been previously added to this equation which accounted in a global way for possible substrate limitations. In the model developed in this work, a carbon substrate limitation appeared explicitly in the production expression. Assuming a sigmoidal variation with time of specific growth rate previously validated, the new production model matched well the entire experimental production kinetics. It has been successfully tested for a wide range of nitrogen supplementations, i.e. from an almost total coupling between growth and production for largely supplemented media, to a high decoupling in case of few available nitrogen. Since all the parameters of this model have an obvious biologic meaning, it may be an unvaluable tool for the comprehension of the phenomenon. The model accounted also well for the variation of the specific production rate versus specific growth rate, avoiding the noise due to the direct differentiation of experimental data. PMID- 11397466 TI - Reduced inhibition of enzymatic hydrolysis of steam-pretreated softwood. AB - Softwood constitutes the main source of lignocellulosic material in Sweden which can be used for ethanol production from renewable resources. To make the biomass to-ethanol process more economically feasible, it is preferable to include the sugar-rich prehydrolysate, i.e. the liquid obtained after the pretreatment step, in the enzymatic hydrolysis of the solid fraction. This study shows that the prehydrolysate inhibits cellulose conversion in the enzymatic hydrolysis step. When the prehydrolysate was included in the enzymatic hydrolysis, the cellulose conversion was reduced by up to 36%. However, this inhibition can be overcome by fermentation of the prehydrolysate prior to enzymatic hydrolysis. PMID- 11397467 TI - Angiogenesis and leukemia. PMID- 11397468 TI - Two booster dose hepatitis B virus vaccination in patients with leukemia. AB - The aim of this study was to interpret the antibody response to hepatitis B (HB) vaccination following a two booster dose schedule in 94 acute lymphoblastic leukemia (ALL) patients. All patients were between 1-16 years of age with negative hepatitis B virus (HBV) serology and normal hepatic function. Fifty patients were vaccinated with Engerix B vaccine, and 44 patients were vaccinated with GenHevac B vaccine, with a schedule of 0, 1, 6 and 0, 1, 2, as well as booster doses, in 12 and 6 months respectively. A second booster was given as a fifth dose to 16 unresponsive patients in each vaccine group, 3 and 6 months after the first booster for Engerix B and GenHevac B vaccines respectively. Dosage was 20 microg HbsAg for all patients. Seroconversion rates with protective level antibody were 35.1% (n=33/94). The figures were 32.1% (n=16/50) and 38.6% (n=17/44) for Engerix B and GenHevac B vaccines, respectively. Seroconversion rate in patients younger than 10 years old was found to be higher (39.11%) than older patients (24%), but this was not statistically significant. This study indicates that one third of the leukemic children undergoing maintenence chemotherapy responded to HB vaccine with protective titers of anti-HBs. We recommend HB vaccination especially in developing countries. PMID- 11397469 TI - Interaction of cytosine arabinoside and lovastatin in human leukemia cells. AB - Cytosine arabinoside (ara-C) is widely used for the treatment of leukemias and displays significant toxicities. Lovastatin, an HMG-CoA reductase inhibitor, is extensively used to treat hypercholesterolemia. To determine whether lovastatin could augment ara-C's activity we have examined their effects in the human erythroleukemia K562 cell line and the ara-C resistant ARAC8D cell line. A synergistic interaction between the two drugs was found. We have demonstrated that the interaction does not occur at the level of RAS but may involve lovastatin's effect of downregulating MAPK activity and preventing ara-C-induced MAPK activation. These studies represent the first description of a potentially beneficial interaction between lovastatin and ara-C that could be applied to the treatment of human leukemia. PMID- 11397470 TI - The widening role of statins: RAS signal transduction and drug-induced cytotoxicity in human leukemia: a commentary to 'interaction of cytosine arabinoside and lovastatin in human leukemia cells'. PMID- 11397471 TI - Treatment of patients with myelodysplastic syndrome with amifostine. AB - The efficacy and toxicity of amifostine (300 mg/m(2) three times a week for three consecutive weeks for a maximum of six courses) was evaluated in 12 patients with primary myelodysplastic syndromes. Dose escalation up to 400 mg/m(2) was allowed to patients who failed to respond. Hemoglobin concentration was increased > or = 1.5 g/dl in two (18%) of the 11 anemic patients. These two patients obtained transfusion independence for 20 weeks. Reticulocyte counts and ANC increased > or = 50% of baseline in four (44%) of the nine patients with reticulocytopenia and in three (25%) of the 12 neutropenic patients. Platelet count increased in three (50%) of the six patients with thrombocytopenia. Progenitor growth of CFU-GMs and BFU-Es improved in 8/12 patients. No major side effects were observed. In conclusion amifostine is well tolerated and can promote the growth of primitive hematopoietic progenitors and ameliorate the cytopenias in MDS patients. PMID- 11397472 TI - The final phase in acute myeloid leukaemia (AML): a study of cause of death, place of death and type of care during the last week of life. AB - The aim of this study was to increase the knowledge of the final phase in acute myeloid leukaemia (AML), a period which entails many complex medical and psychosocial decisions. Data on cause and place of death were gathered through a retrospective review of medical and nursing records of 106 patients with AML who had died during 1995-1997. We focused on the actual phase of the disease and to what extent the patients were prescribed palliative care. With increased knowledge of the dying process in AML there are options to discuss which approach would be the most preferred final phase for an AML patient. PMID- 11397473 TI - Palliative care and induction therapy: a complimentary approach to the treatment of acute myeloid leukemia. PMID- 11397474 TI - Recent improvements in outcome for elderly patients with de novo acute myeloblastic leukemia. AB - A retrospective analysis was performed on 263 consecutive patients aged over 60 with de novo acute myeloid leukemia (AML) diagnosed in a single institution between 1979 and 1998. Eighty-nine patients (33%) received only palliative treatment, while 174 patients (67%) were treated with different intensive chemotherapy regimens. The 5- and 10-year overall survival (OS) for the whole series was 7.7+/-1.2 and 4.3+/-1.6%, respectively. For patients receiving chemotherapy, OS was 10.5+/-2.5 and 7+/-2.6%, while for those patients receiving supportive treatment it was 1.1+/-1.1 and 0%, respectively (P=0.002). Within the group of patients receiving chemotherapy, the complete remission (CR) rate was 46%; treatment failure rate was 54% (36% due to treatment-related mortality and 18% due to resistant disease). Variables influencing CR rate were FAB subtype, CD7 positivity, chemotherapy regimen, creatinine level, hepatomegaly, and period of diagnosis. Median disease-free survival (DFS) duration was 8.4 months with a probability of being disease-free at 10 years of 10+/-5%. There were no significant differences in DFS according to age. According to the period of diagnosis (1979-1986 vs. 1987-1998), improvements in the CR rate (27 vs. 56%, P=0.0002), and OS (10.9 vs. 15.7 months, P=0.0007) were observed. This large single-center study of unselected de novo AML elderly patients substantiates the progressive improvement achieved in the management of elderly patients with AML, probably due to an improvement in supportive care and the administration of conventional induction chemotherapy. PMID- 11397475 TI - Incidence, clinical characteristics and early treatment outcome in Indian patients of childhood acute lymphoblastic leukemia with ALL-1 gene rearrangement. AB - In a series of 185 patients (median age 7 years) of acute lymphoblastic leukaemia (ALL) from India, the overall incidence of ALL-1 gene rearrangement using the Southern blot technique was 11.4% (21/185). The incidence amongst the infants (age < or = 1 year, 70%) was significantly higher when compared to patients > 1 - < or = 10 years (7.4%, P = 0.00001) as well as > 10 years old (9.3%, P = 0.0001). ALL-1 gene rearrangement was associated with significantly higher WBC count (P = 0.01) and CD10 negativity (P = 0.00000001). Complete remission (CR) and relapse rates in 98 patients evaluable for response to therapy on a uniform therapy protocol was independent of ALL-1 gene status. PMID- 11397476 TI - CD8+ T cells in large granular lymphocyte leukemia are not defective in activation- and replication-related apoptosis. AB - Persistent lymphocytosis in large granular lymphocyte leukemia (LGL) may result from defects in activation- or Fas crosslinking-induced cell death. Here we show that Fas crosslinking and CD3 activation causes apoptosis of in vitro activated CD8 T cells, but not of freshly isolated CD8 T cells. Death was partially blocked by a neutralizing antibody to FasL. Inhibition of metalloproteinase-mediated FasL solubilization significantly potentiated induction of cell death. Furthermore, CD3 plus CD28 stimulation resulted in telomeric erosion in LGL cells, and ultimately proliferation ceased. Together, these data indicate that activation- and proliferation-related cell death mechanisms are functional in LGL cells. PMID- 11397477 TI - Angiogenesis in B-chronic lymphocytic leukemia. PMID- 11397478 TI - Interferon-alpha therapy increases type I insulin-like growth factor receptors expression on lymphoid cells from patients with chronic myelogenous leukemia. AB - Aiming to verify if insulin-like growth factor type I and its receptor (IGF-IR) are implicated on pathophysiology of chronic myelogenous leukemia (CML), we studied 35 patients with CML in chronic phase at diagnosis or during interferon alpha (IFN-A) or hydroxyurea treatments. Cytometry flow analysis and reverse transcription PCR (RT-PCR) molecular assay for IGF-IR expression on peripheral blood cells from CML patients diagnosed didn't show statistical differences from the control group. Hydroxyurea treated patients had lower expression of IGF-IR in granulocytes, lymphocytes and monocytes (P<0.01). We found statistical higher percentage of T and B lymphocytes positive for IGF-IR on IFN-A treated patients (P<0.001). Also an increase of IGF-IR mRNA expression could be detected in this group when compared with patients in hydroxyurea therapy (P<0.05). Our study suggest that IGF-IR is not directly implicated on CML installation and that the increased expression of IGF-IR on lymphoid cells of IFN-A treated patients could contribute to the immune recognition of malignant cell clone by enhancing immunocompetent cell proliferation and action. PMID- 11397480 TI - To: the chief resident. PMID- 11397479 TI - Combined histologic and molecular features reveal previously unappreciated subsets of lymphoma in AKXD recombinant inbred mice. AB - Hematopoietic neoplasms developing in AKXD recombinant inbred, NFS.V(+) and ICSBP knockout mice were assessed using morphologic, cytologic and molecular criteria that relate these disorders to human lymphoma and leukemia. Lymphoma types included precursor T-cell and B-cell lymphoblastic, small lymphocytic, splenic marginal zone, follicular, and diffuse large cell (DLCL). In addition to previously defined subtypes of DLCL composed of centroblasts or immunoblasts, two additional subtypes are defined here: lymphoblastic lymphoma like (LL) and lymphoma characterized by a histiocytic reaction (HS). DLCL(HS) were distinguished from true histiocytic lymphomas by the presence of clonal Ig gene rearrangements. PMID- 11397481 TI - Surgical margins in breast cancer: how wide? How accurate? How big? Does it make a difference? PMID- 11397483 TI - Bleeding diverticulosis. PMID- 11397482 TI - Coronary artery bypass. PMID- 11397484 TI - Sarcoma. PMID- 11397485 TI - Comparison of costs of immediate vs delayed breast reconstruction: part 3 of the 6-part series on current concepts in breast reconstruction. PMID- 11397486 TI - Lung volume reduction. PMID- 11397487 TI - Controversy in cryptorchidism: the value of laparoscopy and fertility outcomes. PMID- 11397488 TI - Basics of internet search; so many websites, so little time. PMID- 11397489 TI - Crohn's disease: current concepts in diagnosis and treatment. PMID- 11397491 TI - An incidental paracaval mass. PMID- 11397490 TI - Laparoscopic management of Crohn's disease. PMID- 11397492 TI - Laparoscopic cholecystectomy in patients with early cirrhosis. AB - Traditionally, cholecystectomy in cirrhotic patients has been reserved for patients with severe biliary disease, because of the high morbidity and mortality in cirrhotic patients undergoing this procedure. Laparoscopic cholecytectomy (LC) was originally contraindicated in cirrhotic patients because of the associated portal hypertension and coagulopathy. This study examined the safety of LC in Child's class A patients.A review was conducted of all patients with cirrhosis who underwent cholecystectomy at our hospital between 1990 and 1998.Fifteen patients with cirrhosis had their gallbladder removed laparoscopically during that time period. All patients were Child's class A. The average age was 59 (range, 36-85). The operative indications included acute cholecystitis (5 patients), biliary pacreatitis (4 patients), biliary colic (5 patients), and cholangitis (1 patients). Six patients had known cirrhosis, and 9 were examined intraoperatively. The average operative time was 105 minutes. None of the patients required a blood transfusion. No intraoperative or postoperative complications occurred. No deaths occurred. Postoperative stay was 3 days or less in all but 3 patients.These results compare favorably to other published studies from outside of the United States. Based on our findings, we believe LC can be performed safely in patients with class A cirrhosis. PMID- 11397493 TI - Successful surgical treatment of spontaneous coronary artery dissection. AB - To present a case of spontaneous coronary artery dissection (SCAD). Spontaneous coronary artery dissection is rare and often difficult to surgically repair. Additionally, diagnosis by coronary angiography is uncommon. We present a case of SCAD in a postpartum woman who underwent successful surgical correction of the left anterior descending artery. Such surgical intervention in cases similar to ours is critical for survival. PMID- 11397494 TI - The use of the harmonic scalpel to reduce morbidity during open total abdominal hysterectomies with bilateral salpingo-oophorectomy (TAH/BSO). AB - The ultrasonic scalpel (USS) is a hemostatic surgical instrument used for incising and dissecting tissues. It works by using ultrasonic waves to denature collagen forming a coagulum. This study was designed to examine the effectiveness of the ultrasonic scalpel in open total abdominal hysterectomies with bilateral salpingo-oophrectomies.This study was a nonrandomized study, with 16 cases of total abdominal hysterectomy with bilateral salpingo-oophorectomy being done with the ultrasonic scalpel, and 21 cases being done with the traditional methods of dissection and ligation. The 2 groups were compared in terms of intraoperative blood loss, time, and cost, as well as in terms of postoperative complication and length of stay.Demographically, no significant differences in age or weight existed among the patients in the 2 operative groups. (p = 0.71, p = 0.64, respectively). A statistically significant reduction did occur in the estimated blood loss with the use of the harmonic scalpel (p = 0.05). No significant differences existed between the 2 patient groups in terms of operative time (p = 0.77), intraoperative costs (p = 0.11), or length of hospital stay (p = 0.45). However, 52% of the patients in the traditional method group suffered from postoperative complications that included acute myocardial infarction (N = 1), ileus (N = 5), urinary retention (N = 1), urinary incontinence (N = 1), respiratory failure (N = 1), and failure to thrive (N = 1). Only 25% of the patients undergoing the USS resection suffered from complications. These complications included urinary retention (N = 2), respiratory failure (N = 1), and ileus (N = 1).A statistically significant reduction occurred in estimated blood loss with the use of the USS (p < 0.05). There is also decreased intraoperative time, as well as decreased postoperative complications and length of stay in this same group; however, these differences were not statistically significant. Also, more complications are reported in the traditional resection group (62% compared with 31%). It would be beneficial to accrue a higher patient population and study the use of the USS in this procedure as well as other open abdominal procedures. PMID- 11397495 TI - Will transcatheter embolotherapy replace surgery in the treatment of gastrointestinal bleeding?(2)(2). AB - Superselective angiography and transcatheter embolization (SATE) have produced mixed results in the treatment of upper gastrointestinal bleeding (UGIB). The use of SATE to treat hemorrhage distal to the ligament of Treitz has been approached with trepidation because of limited collateral blood supply to the gastrointestinal tract beyond the duodenum. Ischemic bowel injury rates of 15% to 35% are reported when SATE is used to treat lower gastrointestinal bleeding (LGIB). Recent improvements in coaxial catheter design and embolic agents and greater expertise of personnel performing interventional angiographic procedures have decreased the risks of SATE in the lower gastrointestinal tract. The purpose of this study is to review our experience with SATE in the management of both UGIB and LGIB. %METHODS:This is a retrospective cohort study of patients undergoing SATE between January 1996 and November 1998. All SATE was performed in an angiographic suite with the latest generation coaxial catheters and embolic agents. Patient characteristics and outcome measures were noted. Efficacy of SATE was determined by analyzing the following outcome measures: initial technical success, rebleeding rate at less than 30 days post-SATE, the total rebleeding rate post-SATE, the number of patients requiring surgery despite SATE, and the complication rate. Patients undergoing SATE for LGIB were clinically monitored for development of ischemic complications.Of 523 admissions for UGIB and LGIB, 35 patients (6.7%) underwent SATE as definitive therapy for control of gastrointestinal hemorrhage (25 UGIB and 10 LGIB). Mean age of treated patients was 67 years; 49% had greater than 3 significant systemic comorbidities. Initial control of bleeding was achieved in 23/25 UGIB and 10/10 LGIB patients. Rebleeding at less than 30 days occurred in 5/23 UGIB and 2/10 LGIB patients. Rebleeding at greater than 30 days occurred in 2 additional UGIB patients. No known duodenal stenosis or ischemic bowel injury occurred in patients undergoing SATE for UGIB or LGIB, respectively, during the follow-up period.SATE is an excellent therapeutic method for GI bleeding in selected patients, and definitive control is frequently afforded. In the remainder, SATE may allow temporary control and resuscitation of the unstable patient. SATE appears to be as safe and effective for LGIB as it is for UGIB. Further studies are needed to better define the role of SATE for GI bleeding. PMID- 11397496 TI - Practical use of the Palm Pilot for surgical residents. PMID- 11397498 TI - Mothers of children with cleft palate undergoing speech intervention change communicative interaction. AB - INTRODUCTION: Natural learning must include language learning relationships that provide natural support for communication and language learning. OBJECTIVE: To find out if including the mother as an active participant during speech therapy sessions would improve the communicative style and mode of the interaction of the mothers with their cleft palate children. MATERIALS AND METHODS: Fifty-nine children with cleft palate and their mothers were included in the study group. The patients were divided into two groups randomly. Patients received the same treatment. Twenty-eight of the children were included in the control group. They participated in small working groups comprising the speech pathologist and two children. Thirty-one of the children were included in the experimental group. In this case, the mothers of the children were also included as active participants. The mothers of the patients from the two groups were assessed at the beginning and end of the speech therapy period to find out their style and mode of interaction. Pre- and post-data of the mothers from both groups were compared. RESULTS: Eighty-nine per cent of the mothers of the experimental group modified their patterns of interaction. In contrast, only 19% of the mothers of the control group modified their style and mode of interaction. A Fisher exact test demonstrated that the frequency of mothers from the experimental group that modified their style and mode of interaction was significantly greater as compared to the number of mothers from the control group that were able to modify their style and mode of interaction. CONCLUSIONS: Mothers of children with cleft palate and accompanying language delay modify their communicative style and mode of interaction through active participation in speech therapy. PMID- 11397497 TI - The presence of a widened vestibular aqueduct and progressive sensorineural hearing loss in the branchio-oto-renal syndrome. A family study. AB - OBJECTIVE: A new large family with the BOR syndrome is reported with special reference to the presence of a widened vestibular aqueduct and a progressive sensorineural component in the mixed hearing loss. A review of the BOR literature of 184 patients is given. SETTING: University Hospitals. RESULTS: A BOR family with 17 affected members was studied. Fourteen of those 17 were still alive and 12 of those cooperated in this clinical study. Detailed radiological studies showed in three out of 12 affected family members a widened vestibular aqueduct and progressive sensorineural hearing loss. This raises the question whether there is a true correlation or whether those are coincidental. CONCLUSION: In our family with the Branchio-Oto-Renal syndrome, a widened vestibular aqueduct and progressive hearing loss is found in a few affected family members. Imaging of the temporal bones and long-term audiometric follow-up could help to reveal whether the widened vestibular aqueduct is the cause for the progressive hearing loss. PMID- 11397499 TI - Thyroid ala cartilage reconstruction in neonatal subglottic stenosis as a replacement for the anterior cricoid split. AB - Cricoid split is the procedure of choice in neonatal subglottic stenosis in many paediatric institutions. However, the post-operative care of these patients is a concern given the potentially lethal complications which can occur, in particular, self-extubation. We have been using the thyroid ala cartilage (TAC) reconstruction, a proven technique, as an alternative treatment for this disorder to avoid these complications. The purpose of this study was to compare the results of the TAC reconstruction for this patient population with the results of the cricoid split procedure previously reviewed at our own institution. Between January 1995 and December 1999, 17 patients that underwent the TAC reconstruction for neonatal subglottic stenosis were retrospectively reviewed. Of the 17 patients, two patients required tracheotomy prior to discharge from hospital. Fifteen patients were discharged from hospital with a safe airway without tracheotomy. There were only two minor complications. Compared with the cricoid split study, the success rate with TAC reconstruction was higher (88 vs 83%) and the major complication rate was lower (0 vs 9%). Factors associated with failed procedures may include prematurity, low birth and surgical weight and presence of severe GER. Those that failed required much greater resources in terms of post operative care and length of hospital stay. The TAC reconstruction has replaced cricoid split in treatment of neonates with subglottic stenosis at our institution. PMID- 11397500 TI - Vocabulary acquisition rate after pediatric cochlear implantation and the impact of age at implantation. AB - OBJECTIVE: studies of early vocabulary development after pediatric cochlear implantation show growth rates that approach normality. Do these growth rates continue to rise over time and, therefore, allow a 'catch up' with ideal scores for age, or do they decline after an initial peak. Could age at implantation be a decisive factor in that process? DESIGN: retrospective study (mean follow-up 4 years). PATIENTS: pre-lingually deaf children implanted between 1988 and1999, who serially performed Peabody Picture Vocabulary Test-Revised (PPVT), (37 patients) and Expressive One-word Picture Vocabulary Test-Revised (EOWPVT), (35 patients). OUTCOME MEASURES: the mean rates of age equivalent scores were determined for the whole follow-up period and analyzed further for two post-implant periods (the two halves of follow-up duration of individual patients). After sub-grouping by age at implantation (younger or older than 5 years old), the same analysis was executed for each subgroup. RESULTS: the mean EOWPVT rate of the earlier period was higher than that of the later period (1.33 vs. 0.67, P<0.01) and the mean PPVT rate of the earlier period was higher than that of the later period (0.72 vs. 0.5). The latter difference was not statistically significant (P>0.05). Within subgroups by age at implantation, the PPVT mean rates were stable for younger implanted patients (0.56 for both periods) and dropped for the older implanted sub-group (0.87-0.43, P>0.05). The EOWPVT mean rates declined significantly for the older patients group (1.72-0.55, P<0.01) but insignificantly for the younger patients (0.99-0.77, P>0.05). CONCLUSIONS: vocabulary acquisition rates decline in the post-implantation period. This is more pronounced with older implanted children and the EOWPVT rates. This information on the time course development of vocabulary after implantation would be valuable in counseling and planning habilitation in addition to candidate selection. PMID- 11397501 TI - Infrared transtympanic temperature measurement and otitis media with effusion. AB - Body temperature is an important clinical measurement. It can be estimated by sublingual, rectal, axillary or skin measurement. Pulmonary artery temperature measurement is the gold standard in the research setting, but is obviously inapplicable for routine clinical use. Infrared estimation of core temperature at the tympanic membrane has been proposed as an effective and accurate method. We investigate whether the common condition of childhood otitis media with effusion (OME) interferes with this mode of body temperature measurement. Ninety-five children undergoing grommet tube insertion were assessed preoperatively by axillary and transtympanic routes. The presence or absence of middle ear fluid was noted at the time of surgery. Analysis of the data showed no influence of OME on the measurement of body temperature by the transtympanic route compared with axillary measurement. PMID- 11397503 TI - Cochlear implantation in children with large vestibular aqueduct syndrome and a review of the syndrome. AB - OBJECTIVE: Children with Large Vestibular Aqueduct Syndrome (LVAS) frequently develop speech and language skills prior to deterioration of their hearing. Operations designed to halt the progression of hearing loss have largely failed so the question of Cochlear Implantation in these children has arisen. It had been suggested that there would be technical difficulties in implanting these patients and, therefore, there had been an initial reluctance to proceed to implantation. The aim of the present paper is to assess surgical and functional outcomes in implanted children with LVAS and review the related literature. MATERIAL AND METHODS: From the 170 children assessed by MRI in the Nottingham Paediatric Cochlear Implant Programme, seven (4%) were identified as having LVAS. Four of these children were implanted and had at least 12 months follow up. Two of the children are on the waiting list for implantation and one child was not implanted because of absence of the cochlear nerve. Operative findings, complications and outcome measures were recorded. The auditory skills of the children were assessed before implantation and 1 year following implantation. A literature search was done to identify other series with experience in implanting children with LVAS. RESULTS: Full insertion of the electrode array was achieved in all our cases. After cochleostomy two patients experienced a mild CSF leak that was easily controlled by the muscle graft. On the first day post-operation two patients were nauseous and one had an episode of vomiting, however, all were discharged within 24 h of surgery. Initial outcome measures at 12 months post implantation were encouraging showing significant progress in children's auditory skills. CONCLUSIONS: The results of the present study and the review of the literature suggest that LVAS is not a contraindication to implantation as initial concerns about severe perilymph leaks and surgical complications have proved to be unfounded. The post-operative progress of these children in listening skills also suggest that these children are suitable for cochlear implantation PMID- 11397502 TI - Predicting outcome in pediatric coin ingestion. AB - OBJECTIVE: To determine the relationship between coin size, coin location, patient age, and patient weight and likelihood of coin passage through the esophagus following pediatric coin ingestion. A secondary objective is to test the hypothesis that coin denomination can be determined based on radiographic appearance. METHODS: A retrospective review was performed of all children seen and evaluated for coin ingestion at a single institution over a 25-month period. Outcome measures included the number of coins that were retained in the esophagus, and the number that passed. Various factors were assessed for their predictive value in judging outcome in coin ingestion cases. RESULTS: Nineteen percent of patients (15/79) in the study passed their ingested coins. Coin denomination could be accurately determined on every patient that had a standard AP or lateral X-ray film. These findings were marked when compared with the lack of reliability of history in determining coin denomination. Patients who passed coins were as a group older (4.6 vs. 3.2 year, P=0.04), but did not differ significantly by weight (19.5 vs. 15.4 kg, P=0.07) from those that retained the coins. Coins located at the gastroesophageal junction had a significantly higher passage rate than coins located elsewhere in the esophagus (89 vs. 8.2%, P<0.01). Coin size was not predictive of coin passage (P=0.7 by chi(2)). CONCLUSIONS: Radiographic assessment of coin denomination is reliable, but in this study could not be used to predict coin passage. Patient age and coin location at the gastroesophageal junction, however, do correlate with this event. PMID- 11397504 TI - Embedded sharp metallic spring in esophagus. AB - Less than 1% of esophageal foreign bodies are irretrievable by endoscopic techniques. These cases require esophagotomy for removal. The impacted metallic spring with sharp ends (in opposite direction) is reported as esophageal foreign body which required esophagotomy after two failed endoscopic removals. The difficulties encountered during endoscopic maneuver, peculiar shape of foreign body and mode of impaction are discussed. PMID- 11397505 TI - Otolaryngologic manifestations of Rosai-Dorfman Disease. AB - PURPOSE: To describe an unusual head and neck occurrence of Rosai-Dorfman Disease (RDD) and to review the otolaryngologic manifestations of this rare entity. METHODS: A case presentation with review of the literature describing Rosai Dorfman Disease and its head and neck involvement. SETTING: A tertiary care, urban children's hospital. RESULTS: This is the first description, to the best of our knowledge, of RDD [Sinus Histiocytosis with Massive Lymphadenopathy (SHML)] involving bilateral external auditory canals and middle ear in a 12 year old patient previously diagnosed with 'asthma' and hearing loss. This patient also had extensive involvement of the tracheobronchial tree. Representative clinical, radiographic and histological findings are presented. Its etiology, diagnosis and management are also reviewed. CONCLUSION: This is the first reported case of middle ear and external auditory canal involvement of RDD in a patient with hearing loss and associated tracheobronchial lesions. RDD, although rare, may be considered in the differential diagnosis of unusual histiocytic lesions of the external auditory canal, especially with findings of similar or comparable lesions involving the respiratory tract. Confirmation is with identification of emperipolesis and appropriate immunohistochemical staining (S-100 positive, CD-68 positive and CD-1a negative). Intervention is recommended in cases where there is increased risk of mortality, as in severe obstruction of the tracheobronchial tree. Otherwise, since these lesions are self-limiting, the patients can be observed closely. PMID- 11397506 TI - Current research on respiratory viral infections: Third International Symposium. PMID- 11397507 TI - Higher levels of HIV DNA in memory and naive CD4(+) T cell subsets of viremic compared to non-viremic patients after 18 and 24 months of HAART. AB - The degree of infection of memory and naive CD4(+) T cells in patients treated with HAART and with durable undetectable or detectable viral load in plasma was evaluated. The following two groups of patients were analyzed cross-sectionally: (i) patients with undetectable HIV RNA plasma levels during follow-up (responders); (ii) patients with no reduction or with rebound in HIV RNA levels during treatment (non-responders). Patients were examined following 6, 12, 18 and 24 months of HAART, respectively, by quantifying: (i) plasma HIV RNA load; (ii) CD4(+) T cells; (iii) memory and naive CD4(+) T cells; (iv) HIV DNA levels in memory and naive CD4(+) T cells. HIV RNA plasma levels were significantly higher in non-responders vs responders at each time point (P<0.02), while CD4(+) T cell counts as well as memory and naive CD4(+) T cell levels were comparable in both viremic and non-viremic patients. However, higher HIV DNA values were observed in both memory and naive CD4(+) T cells of non-responders vs responders after 18 and 24 months of HAART (P<0.02), suggesting an increased amount of HIV-infected naive CD4(+) T cells and a sustained high degree of infection of memory CD4(+) T cells. Immunological reconstitution following HAART might potentially be hampered in viremic patients despite the absolute increase in CD4(+) T cell counts. PMID- 11397508 TI - Exploring the role of the 5'-position of TSAO-T. Synthesis and anti-HIV evaluation of novel TSAO-T derivatives. AB - Various analogues of the anti-HIV-1 agent TSAO-T, [1-[2',5'-bis-O-(tert butyldimethylsilyl)-beta-D-ribofuranosyl]thymine]-3'-spiro-5"-(4"-amino-1",2" oxathiole-2",2"-dioxide) have been synthesized in which the 5'-TBDMS group has been replaced by alkyl-, alkenyl- or aromatic ether groups, substituted amines, carbamoyl or (thio)acyl groups. The compounds synthesized were evaluated for their inhibitory effect on HIV-1 and HIV-2 replication in cell culture. Replacement of the 5'-TBDMS group by an acyl, aromatic or a cyclic moiety markedly diminish or even eliminate the anti-HIV activity. However, the presence at that position of an alkyl or alkenyl chain, partially retain antiviral activity. These observations suggest that the 5'-TBDMS group of the TSAO molecule plays a crucial role. PMID- 11397509 TI - Antiviral activity of an extract derived from roots of Eleutherococcus senticosus. AB - A liquid extract from Eleutherococcus senticosus roots inhibited the productive replication of human rhinovirus (HRV), respiratory syncytial virus (RSV) and influenza A virus in cell cultures infected with these viruses, all of which belong to the RNA type viruses. Analysis of virus production after treatment of the infected cells using plaque-reduction assays showed a strong antiviral activity of the Eleutherococcus extract. In contrast, no effect was detected using the same protocol for cells infected with the DNA viruses, adenovirus (Adeno 5) or herpes simplex type 1 virus (HSV 1). Pre-treatment of cells did not inhibit either virus adsorption or virus replication. The results of the study demonstrate that the Eleutherococcus extract inhibited the replication of all RNA viruses studied so far. This antiviral activity remained stable under the conditions used for drug preparation and storage. PMID- 11397511 TI - Luteolin-inhibited arylamine N-acetyltransferase activity and DNA-2-aminofluorene adduct in human and mouse leukemia cells. AB - N-Acetyltransferase enzyme is an important enzyme in the first step of arylamine compounds metabolism. Luteolin has been shown to exit antibacterial and antineoplastic activity. The purpose of this present study is to evaluate the question of whether luteolin could affect arylamine N-acetyltransferase (NAT) activity and DNA-2-aminofluorene adduct formation in human (HL-60) and mouse (L1210) leukemia cells. By using HPLC, N-acetylation of 2-aminofluorene was determined. Luteolin displayed a dose-dependent inhibition to cytosolic NAT activity and intact human and mice leukemia cells. Time-course experiments showed that N-acetylation of 2-aminofluorene measured from intact human and mice leukemia cells were inhibited by luteolin for up to 24 hours. Using standard steady-state kinetic analysis, it was demonstrated that luteolin was a possible uncompetitive inhibitor to NAT activity in cytosols. The DNA-2-aminofluorene adduct formation in human and mouse leukemia cells were inhibited by luteolin. This report is the first demonstration to show that luteolin affects human and mice leukemia cells NAT activity and DNA-2-aminofluorene on adduct formation. PMID- 11397510 TI - Lack of protective action of cysteine against the fetotoxic effect of monocrotaline. AB - Monocrotaline (MCT), a pyrrolizidine alkaloid present in Crotalaria species, has hepatotoxic, nephrotoxic, pneumotoxic and fetotoxic effects. However, the toxic effects of exposure to MCT in adult rats can be prevented by cysteine. Thus, the present study was conducted to evaluate the possible prevention by cysteine of the toxic effects of MCT on pregnant rats. Thirty-six pregnant rats were used. The females in the experimental groups were fed ration containing 0.02% MCT, 0.02% MCT + 1% cysteine, or 1% cysteine from day 6 to day 21 of pregnancy; the control group was fed only common ration for the same period of time. All rats were killed on day 21 of pregnancy and their blood was collected for determination of liver and kidney function. General toxicity to pregnant dams was assessed. Fetuses were removed by caesarian section and embryofetotoxic parameters were examined. Results showed impaired body weight gain in rats fed MCT, with or without cysteine supplementation. Plasma levels of AST, ALT, LDH, GGT, urea and creatinine were increased in MCT animals compared to controls. The pathology study revealed lesions only in dams from the MCT group. The weights of the placentas and fetuses of the MCT and MCT + cysteine groups were significantly lower than those of the control group. Thus, the present data suggests some protective action of 1% of cysteine in ration against the toxic effects of MCT on the dams but not on the litter. PMID- 11397512 TI - Assessing cyanogen content in cassava-based food using the enzyme-dipstick method. AB - The cyanogen content of 35 cassava-based foods was determined using the enzyme dipstick method. The analyses showed presence of residual cyanogens in these products, and they ranged from 2 to 88 mg HCN equivalent/kg. Foodstuff prepared from grated cassava roots exhibited a lower level of cyanogen content (2-31 mg HCN equivalent/kg) compared with those prepared as cassava root slices or from cassava flour (28-88 mg HCN equivalent/kg). These results showed that current commercial processing methods did not remove all the cyanogen present from cassava roots and it might be worthwhile re-examining these processing practices. In this respect, the report showed that an overnight treatment of cassava root slices with linamarase and cellulase might facilitate the removal of cyanogens. This study also demonstrated the usefulness of enzyme-dipsticks for assessing cyanogen content in cassava food products. PMID- 11397513 TI - A comparison of whole wheat, refined wheat and wheat bran as inhibitors of heterocyclic amines in the Salmonella mutagenicity assay and in the rat colonic aberrant crypt focus assay. AB - Refined wheat, unrefined whole wheat, and wheat bran were studied for their ability to protect against heterocyclic amines (HCAs) in vitro and in vivo. Wheat bran, which binds HCAs in vitro, as well as refined wheat and unrefined whole wheat, inhibited the mutagenic activities of 2-amino-3-methylimidazo[4,5 f]quinoline (IQ), 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2 amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx) when they were co-incubated and the supernatant (minus grain) was added to the Salmonella assay. The water soluble fraction alone from refined and unrefined wheat, but not bran, also inhibited against these mutagens in vitro. In vivo, AIN-93G diets containing refined wheat or unrefined wheat were examined for their ability to inhibit IQ induced colonic aberrant crypt foci (ACF) in the Fischer 344 rat. A slight increase in the number of AC/ACF (aberrant crypts/ACF) was seen after 16 weeks in rats treated post-initiation with refined wheat (P < 0.05), and fewer foci with two or three aberrant crypts (ACF-2) were found in rats given unrefined whole wheat post-initiation compared with animals treated with the same diet during the initiation phase (P < 0.05). There was no significant difference in the profile of IQ urinary metabolites or excretion of promutagens 0-48 h after carcinogen dosing, and grains had no effect on hepatic cytochrome P4501A1 (CYP1A1), CYP1A2, aryl sulfotransferase or N-acetyltransferase activities; however, a slightly higher UDP-glucuronosyl transferase activity was observed in rats fed unrefined wheat compared with refined wheat diets (P < 0.05). Thus, despite their antimutagenic activities in vitro, only marginal effects were seen with refined and unrefined wheat in vivo with respect to hepatic enzyme activities, carcinogen metabolism and IQ-induced ACF in the rat colon. PMID- 11397514 TI - Uncertainty factors for chemical risk assessment: interspecies differences in the in vivo pharmacokinetics and metabolism of human CYP1A2 substrates. AB - The 100-fold default uncertainty factor is used to convert a no-observed-adverse effect level (NOAEL) from a animal toxicity study, to a "safe" value for human intake. The composite uncertainty factor (100) has to allow for interspecies (10 fold) and interindividual (10-fold) differences in toxicokinetics and toxicodynamics. The aim of the current study was to assess the validity of the interspecies default for toxicokinetics (4.0) for each of the test species (dog, rabbit, rat and mouse), using published data for compounds eliminated by CYP1A2 in humans (caffeine, theobromine, theophylline and paraxanthine). An analysis of the published literature showed that the absorption, bioavailability and route of excretion were generally similar between humans and the test species, for each probe substrate. However, interspecies differences in the route of metabolism, and the enzymes involved in this process, were identified. The magnitude of difference in the internal dose, between species, showed that values for the mouse (10.6) and rat (5.4) exceed the 4.0-fold default, whereas the rabbit (2.6) and dog (1.6) were below this value. This work supports the need to replace the generic default factors by a compound-related value derived from specific, relevant, quantitative data; this would result in more relevant and reliable non cancer risk assessments. PMID- 11397515 TI - Uncertainty factors for chemical risk assessment. human variability in the pharmacokinetics of CYP1A2 probe substrates. AB - A 100-fold uncertainty factor is used to derive acceptable daily intakes for compounds causing thresholded toxicity. The 10-fold factor for human variability can be further subdivided into two factors of 10(0.5) (3.16) to allow for toxicokinetics and toxicodynamics. The validity of the human kinetic subfactor has been analysed in relation to CYP1A2 metabolism using published in vivo pharmacokinetic parameters selected to reflect chronic exposure (metabolic and total clearances and area under the plasma concentration-time curve: CLm, CL and AUC) and acute exposure (the peak plasma concentration, C(max)). The variability in CYP1A2 activity in healthy adults, based on data after oral and intravenous dosage (CLm, CL and AUC), ranged from 34 to 42%. The variability in C(max) was 21%. The default kinetic factor of 3.16 would cover at least 99% of the healthy adult population, assuming that the data were log-normally distributed, but would give lower protection for some subgroups (pregnant women at term, healthy elderly, patients with liver disease), and was inadequate for neonates. This analysis of in vivo kinetic data for CYP1A2 substrates illustrates the importance of quantifying human variability in specific metabolic pathways, and of identifying potentially susceptible subgroups of the human population, in order to determine the scientific validity of uncertainty factors. PMID- 11397516 TI - The benchmark approach applied to a 28-day toxicity study with Rhodorsil Silane in rats. the impact of increasing the number of dose groups. AB - The OECD study design, aimed at obtaining a no-observed-adverse-effect level (NOAEL), may be suboptimal for deriving a benchmark dose. Therefore the present subacute (28-day) study was carried out to evaluate a multiple dose study design and to compare the results with the common OECD design. Seven groups of 10 female rats each were intragastrically administered corn oil without (controls) or with 50, 150, 300, 450, 600 or 750 mg Rhodorsil Silane/kg body weight/day, once daily (7 days/week) for 4 weeks. From the complete dataset, two subsets were selected, one representing a study design with seven dose groups of five animals (7 x 5 design), the other representing a study design with four dose groups of 10 animals (4 x 10 design). Under the conditions of the present study, the NOAEL for Rhodorsil Silane 198 was assessed at 50 mg/kg body weight/day, based on the data of the 4 x 10 design. The benchmark approach resulted in a benchmark dose of 19 mg/kg body weight/day, based on the data of the 7 x 5 design. Comparison of the results demonstrated that the multiple dose (7 x 5) design led to a more reliable result than the OECD (4 x 10) design, despite the smaller total number of animals. The dose-response analysis showed that at "the NOAEL" the effect on relative spleen weight was larger than 10%, illustrating that at the NOAEL, adverse effects may occur. PMID- 11397517 TI - The absorption, distribution and excretion of 1- and 2-. AB - Human breast milk is rich in 2-palmitoyl 1,3 unsaturated triacyglycerols and during the neonatal period, when milk is the sole source of nutrients, their role could be particularly important. Betapol is a novel triacylglycerol mix resembling human breast milk in its high palmitic acid content and positional distribution. The total fat absorption from Betapol has been shown to be higher than fat from conventional infant milk formulas and closer to human breast milk in infants. However, the relative fate of purified palmitic acid esterified to glycerol in the 1-, 3- and 2-positions in neonatal and young animals has not previously been established. Therefore, the fate of orally administered 1-[1 14C]palmitoyl, 2,3 dioleoyl glycerol ([14C]POO) and 1,3 dioleoyl,2-[1 14C]palmitoyl glycerol (O[14C]PO) was investigated in suckling and weanling rats using liquid scintillation counting of tissues and expired air and whole-body autoradiography. The results obtained indicate that orally administered [14C]POO and O[14C]PO are extensively absorbed from the gut, probably either as palmitic acid or as a palmitoyl glyceride in both suckling and weanling rats. Radioactivity initially concentrated in brown fat with apparent migration to the white fat of weanling rats by 96 h. Levels of 14C were low in blood, brain and other tissues. Excretion of 14C was mainly by expiration of CO(2) (approximately 72% in 96 h), indicating beta-oxidation as a major route of metabolism. Urine and faeces accounted for only approximately 6% of the excreted radioactivity. The design and size of the experiment did not allow tests of statistical significance between the absorption and excretion of OPO and POO to be conducted. However, the absorption, distribution, beta-oxidation and excretion appeared to be similar. PMID- 11397518 TI - The effect of the zeolite clinoptilolite on serum chemistry and hematopoiesis in mice. AB - Zeolites are natural or synthetic crystalline alumosilicates with ion exchanging properties. Supplied in fodder, they promote biomass production and animal health. Our aim was to assess the effects of the natural zeolite, clinoptilolite, on hematopoiesis, serum electrolytes and essential biochemical indicators of kidney and liver function in mice. Two preparations differing in particle size were tested: a powderized form obtained by countercurrent mechanical treatment of the clinoptilolite (MTCp) and normally ground clinoptilolite (NGCp). Young adult mice were supplied with food containing 12.5, 25 or 50% clinoptilolite powder. Control animals received the same food ration without the clinoptilolite. After 10, 20, 30 and 40 days, six animals from each group were exsanguinated to obtain blood for hematological and serum for biochemical measurements as well as to collect femoral bone marrow for determination of hematopoietic activity. Clinoptilolite ingestion was well tolerated, as judged by comparable body masses of treated and control animals. A 20% increase of the potassium level was detected in mice receiving the zeolite-rich diet, without other changes in serum chemistry. Erythrocyte, hemoglobin and platelet levels in peripheral blood were not materially affected. NGCp caused leukocytosis, with concomitant decline of the GM-CFU content in the bone marrow, which was attributed to intestinal irritation by rough zeolite particles. The mechanically treated clinoptilolite preparation caused similar, albeit less pronounced, changes. In a limited experiment, mice having transplanted mammary carcinoma in the terminal stage showed increased potassium and decreased sodium and chloride levels, severe anemia and leukocytosis, decreased bone marrow cellularity and diminished content of hematopoietic progenitor cells in the marrow. The clinoptilolite preparations ameliorated the sodium and chloride decline, whereas the effects on hematopoiesis were erratic. PMID- 11397519 TI - The effect of zinc supply on cadmium-induced changes in the tibia of rats. AB - It has been determined that zinc supplementation (240 microg Zn/ml) during (for 12 weeks) or after (for 2 weeks) cadmium exposure (50 microg Cd/ml for 12 weeks) can prevent the accumulation and toxic action of Cd in the tibia of rats. The exposure to Cd led to disturbances in bone metabolism reflected by changes in the chemical composition of bone and decreased bone mineral density (osteomalacian changes). The Zn supply in conditions of Cd intoxication completely prevented the Cd-induced increase in percentage of water content and decrease in tibia ash weight, ash weight/dry weight, non-org. comp./org. comp., Zn content and concentration. Moreover, Zn partly protected from the decrease in Ca concentration and content, percentage of non-organic components content, Ca/wet weight, Ca/ash weight and Ca/dry weight. Zn administered after Cd exposure partly, but not completely, protected from Cd-induced decrease in percentage of non-organic components content, Ca/wet weight as well as Ca content and concentration. This protective effect on bone was most evident when Zn was administered during Cd exposure. But Zn, independently of the manner of its administration, did not prevent Cd accumulation in the tibia. Our results suggest that Zn supply in conditions of simultaneous exposure can prevent Cd-induced bone loss to some extent, and used after Cd treatment can give therapeutic benefits. PMID- 11397520 TI - Carcinogenesis bioassays: study duration and biological relevance. AB - Criticisms of the scientific value of rodent carcinogenicity bioassays have focused on the arguments that the studies are too long and that most organ specific carcinogenic effects observed in experimental animals have little or no relevance to humans. For example, Davies et al. (Davies, T.S., Lynch, B.S., Monro, A.M., Munro, I.C., Nestmann, E.R., 2000. Rodent carcinogenicity tests need be no longer than 18 months: an analysis based on 210 chemicals in the IARC Monographs. Food and Chemical Toxicology 38, 219-235) concluded that the duration of rodent bioassays should be no more than 18 months, based on their analysis of 210 International Agency for Research on Cancer (IARC) rodent carcinogens in which they report that most chemicals showed "tumorigenic effects" at or before 12 months. However, many of these "tumorigenic effects" reflect the occurrence of a single neoplasm, with most tumors occurring much later in the study. Reliance on a single tumor at an early time point as providing definitive evidence of rodent carcinogenicity is a dangerous practice that could produce both false positive and false negative outcomes. An extensive evaluation of the NTP database reveals that many rodent carcinogens produce later-appearing tumors that would not be detected as statistically significant in a 12-18 month study. Such a shortened duration study would be roughly equivalent to evaluating human cancer in subjects 30-50 years of age, which would result in markedly reduced study sensitivity. In fact, many investigators recommend extending the duration of rodent studies to 30 months or to a true lifetime to increase study sensitivity. We also do not agree with the second conclusion of Davies et al. (2000) that the mode of action of rodent carcinogenesis is sufficiently well understood to justify discounting the majority of organ-specific carcinogenic effects found in these studies. The consequences of performing rodent carcinogenicity studies with inadequate sensitivity, and then discounting most of the carcinogenic effects that are observed will be that potential human carcinogens will not be detected, thus forcing near total reliance on human studies for this purpose. This is not prudent public health policy. PMID- 11397521 TI - Calculation of the intake of three intense sweeteners in young insulin-dependent diabetics. AB - In 1994, European Directive 94/35/CE authorised the use as food additives of five intense sweeteners for which Acceptable Daily Intakes (ADI) were established. The same directive stipulated that member states should organise a monitoring system to determine the consumption of these substances. Diabetic children are normally considered to constitute a group with a high consumption of sweeteners (European Commission, 1998. Report on Methodology for the Monitoring of Food Additives Intake across the European Union. Report of the Scientific Cooperation, Task 4.2 SCOOP/INT/REPORT/2. European Commission Directorate General III, Brussels.). A stepwise approach to the food additive intake in the general population had shown that three of the five authorised intense sweeteners (aspartame, saccharin and acesulfame K) are used at particularly high levels in sugar-free foods and are also very commonly utilised as table-top sweeteners. This paper presents the results of a food intake survey conducted in a group of French, insulin-dependent children in 1997, aimed at estimating the Theoretical Maximum Daily Intake (TMDI) for these three sweeteners and comparing this with the relevant ADI values. A 5 day diary questionnaire was used to estimate the intake of sugar-free, artificially sweetened foods and table-top sweeteners. When assessing the intake of each additive, all sugar-free products were assumed to be sweetened using a single sweetener at its maximum authorised level. This study was performed in five age groups, and based on the mean and 97.5th percentile of the distribution of consumption, demonstrated that it was unlikely that total exposure could rise above the ADI. PMID- 11397522 TI - Behaviour of some organophosphorus and organochlorine pesticides in potatoes during soaking in different solutions. AB - The efficiencies of acidic solutions (radish, citric acid, ascorbic acid, acetic acid and hydrogen peroxide), neutral solutions (sodium chloride) and alkaline solution (sodium carbonate) as well as tap water in the elimination of organochlorine and organophosphorus pesticides from naturally contaminated potatoes were examined. The results indicated that acidic solutions were more effective than neutral and alkaline solutions in the elimination of the organochlorine compounds under investigation, Radish solutions eliminated pesticides completely, except o,p'-DDE (73.1% loss), followed by citric and ascorbic acid solutions. On the other hand, organophosphorus pesticides (pirimphos methyl, malathion and profenofos) were eliminated more by acidic, neutral and alkaline solutions than by organochlorines. The percentage of removal ranged from 98.5 to 100% for pirimphos methyl, 87.9 to 100% for malathion and 100% for profenofos. PMID- 11397523 TI - Reproducibility of contrast-enhanced transrectal ultrasound of the prostate. AB - Transrectal three-dimensional (3-D) contrast-enhanced power Doppler ultrasound (US) is a novel technique for studying possible prostate malignancy. Before studies can be performed to investigate the clinical validity of the technique, reproducibility of the contrast US studies must be proven. Reproducibility of contrast US was studied in 10 patients with biopsy-proven prostate cancer. The studies performed included static investigations and dynamic investigations of the prostate vasculature. All studies were double performed. The assessment of reproducibility was done objectively using a computer program and, subjectively, by visual assessment. The results indicate high reproducibility of static contrast investigations, for both the objective and subjective assessment. The subjective assessment of the dynamic studies was also highly reproducible. The objective assessment of the dynamic contrast studies, however, was less reproducible, mainly due to motion artefact. We concluded that, especially static 3-D contrast-enhanced, power Doppler investigations of the prostate are highly reproducible. PMID- 11397524 TI - Reproducibility of skin characterization with backscattered spectra (12--25 MHz) in healthy subjects. AB - Ultrasonic techniques were developed for quantitative in vivo analysis of skin composition based on measurements of apparent integrated backscatter (IBS) and its frequency dependence (n) between 12.5 and 25 MHz. Parameters were measured at five depths in healthy dermis of the midforearm of 29 volunteers (13 women, 16 men, 20 to 76 years old) on three different days. Reproducibility of measurements was evaluated (standardized coefficients of variation: 7% to 11% for IBS and 9% to 20% for n). Parameter values were significantly influenced by pressure of the ultrasonic probe on the skin, and both room and skin-surface temperatures were correlated to IBS measured in a single subject on 28 days. More precise control of these factors could further improve measurement reproducibility and sensitivity to skin composition. Significant (p < 0.05) differences of parameter values with respect to region of interest depth, age and gender of subjects were discussed in comparison with dermal composition and offer promise that these parameters could be used to characterize skin modifications. PMID- 11397525 TI - Age-related alterations of cardiac tissue microstructure and material properties in Fischer 344 rats. AB - The cardiac aging process is accompanied by global mechanical dysfunction that reflects increased myocardial stiffness. Accordingly, age-related changes in microscopic material properties of myocardium were delineated with high-frequency ultrasound (US) (30 to 44 MHz) tissue characterization methods for aging Fischer 344 rats at 6 (adult), 18 (aged), and 24 (senescent) months of age. The excised lateral wall of the left ventricle of rats (n = 10 per group) was insonified with a 50-MHz acoustic microscope for determination of integrated backscatter, backscatter coefficient and attenuation coefficient. Histological and biochemical analyses for collagen content and cardiac myocyte diameter were performed. Collagen concentration increased progressively with age, with the greatest increments occurring from 6 to 18 months (38.0 +/- 6.3 to 53.0 +/- 7.1 mg/g dry wt), and leveling off at 24 months (60.0 +/- 7.4 mg/g dry wt). Tissue microscopic material properties also changed progressively from 6 to 24 months of age, as determined by US methods: integrated backscatter increased (-44.7 +/- 1.8 vs. 40.8 +/- 1.9 dB, p < 0.05), attenuation increased (47.1 +/- 5.9 to 65.3 +/- 7.8 dB/cm, p < 0.05), and the backscatter coefficient increased (0.73 +/- 0.16 x 10( 5) to 3.76 +/- 1.6 x 10(-5) cm(-1), p < 0.05), from 6 to 24 months of age in each case. Age-related alterations in indices of cardiac microscopic material properties were closely correlated with the changes in cardiac microstructure. Ultrasonic tissue characterization may prove to be a sensitive tool to monitor changes in the cardiac microstructure, such as increased collagen deposition, that occur within age-related diastolic dysfunction. PMID- 11397526 TI - Spatio-temporal mapping of intracardiac pressure gradients. A solution to Euler's equation from digital postprocessing of color Doppler M-mode echocardiograms. AB - Doppler assessment of intracardiac pressure gradients using the simplified Bernoulli equation is inaccurate in the absence of a restricted orifice. The purpose of this study is to develop a new general method to map instantaneous pressure gradients inside the heart using Doppler echocardiography. Color Doppler M-mode recordings are digitally postprocessed with a software algorithm that decodes flow velocity and fits a bivariate spatio-temporal tensor-product smoothing spline. Temporal and spatial accelerations are then calculated by analytical derivation of the fitted velocity data, allowing solution of both inertial and convective terms of Euler's equation. A database of 39 transmitral inflow and transaortic outflow color Doppler M-mode recordings from 20 patients with a number of cardiac conditions was analysed, along with matched pulsed-wave spectral recordings. A close agreement was observed between the spectral and postprocessed color Doppler velocity values (error = 0.8 +/- 11.7 cm/s), validating the data decoding and fitting process. Spatio-temporal pressure gradient maps were obtained from all studies, allowing visualisation of instantaneous pressure gradients from the atrium to the apex during left ventricular filling, and from the apex to the outflow tract during ejection. Instantaneous pressure differences between localised intracardiac sample points closely matched previously published catheterization findings, both in magnitude and waveform shape. Our method shows that intracardiac instantaneous pressure gradients can be analysed noninvasively using color Doppler M-mode echocardiography combined with image postprocessing methods. PMID- 11397527 TI - Neurovascular coupling in terms of a control system: validation of a second-order linear system model. AB - Neurovascular coupling (NC) adapts cerebral blood flow to cortical activity. Functional transcranial Doppler (f-TCD) investigations revealed a typical time course of evoked blood flow responses with an initial overshoot and a stabilization at a lower, but stable, level. This blood flow reaction can be described in terms of a control-system model. We tested reliability and validity of this new approach using different stimulation paradigms. The P2 segment of the posterior cerebral artery was insonated in 14 healthy volunteers and, subsequently, NC was stimulated using two tests, a checkerboard stimulus of 1 s with different repetition rates and a reading test. Data were analyzed according to algorithms of control-system theory. The reading test was used to measure test reliability and side differences. A second-order linear system can describe blood flow regulation of NC. Different stimulation protocols of the checkerboard test and the related evoked blood flow curves could be described by the same control system model. Further, there were no differences between the checkerboard and reading test nor between right and left side or test and retest of the reading test. NC can be described in a much more detailed manner using control-system analysis. We were able to show that blood flow response due to different visual stimuli follow one common control-system model. Unlike quantification of NC using overshoot, parameters of the control system have smaller SDs, increasing the statistical power and, thereby, usefulness of f-TCD as a diagnostic instrument. PMID- 11397529 TI - Spatial averaging in the beam of a piston transducer. AB - This paper presents a theoretical analysis of the spatially averaged free-field responses of phase-sensitive and phase-insensitive receivers centered in the beam of a harmonically excited piston transmitter. The responses of unfocused circular plane piston receivers are analyzed, and both unfocused and spherically focused piston transmitters are considered. A set of closed-form expressions figures prominently in the analysis. The expressions are based on the Lommel diffraction formulation which is, in turn, based on the Fresnel approximation. Although approximate, the expressions allow for quick and easy estimation of phase sensitive or phase-insensitive unfocused piston receiver responses. It is shown that the spatial averaging effects associated with phase-sensitive and phase insensitive receivers are virtually identical when gamma < or = 0.1, where gamma = b/a is the ratio of receiver radius b to transmitter radius a. In addition, numerical results obtained from the closed-form expressions are compared with previously reported results. The comparisons indicate that the approximate results are valid from the m = 3 maxima forward under the assumption of linear propagation when ka > 58, where k is the circular wave number. Finally, it is pointed out that the closed-form expressions may prove useful in the estimation of the potential for bioeffects associated with diagnostic ultrasound. PMID- 11397530 TI - Sources of error in maximum velocity estimation using linear phased-array Doppler systems with steady flow. AB - Using linear-array Doppler ultrasound (US) transducers, the measured maximum velocity may be in error and lead to incorrect clinical diagnosis. This study investigates the existence and cause of maximum velocity estimation errors for steady flow of a blood-mimicking fluid in a tissue-mimicking phantom. A specially designed system was used that enabled fine control of flow rate, transducer positioning and transducer angle relative to the flow phantom. Doppler machine settings (transducer aperture size, focal depth, beam-steering, gain) were varied to investigate a wide range of clinical applications. To estimate the maximum velocity, a new signal-to-noise ratio (SNR) independent method was developed to calculate the maximum frequency from an ensemble averaged Doppler power spectrum. This enabled the impact of each factor on the total Doppler error to be determined. When using the new maximum frequency estimator, it was found that the effect of transducer focal depth, intratransducer, intramachine, intermachine (that was tested) and beam-steering did not significantly contribute to maximum velocity estimation errors. Instead, it was the dependence of the maximum velocity on the Doppler angle that made, by far, the greatest contribution to the estimation error. Because our maximum frequency estimator took into account the effect of intrinsic spectral broadening, the degree of overestimation error was not as great as that previously published. Thus, the effects of Doppler angle and intrinsic spectral broadening are the chief sources of Doppler US error and should be the focus of future efforts to improve the accuracy. PMID- 11397528 TI - In vitro quantification of flow using continuous infusion of Levovist and pairs of harmonic power Doppler images. AB - To evaluate the potential of harmonic power Doppler to quantify perfusion using a continuous infusion of contrast, two dialysis cartridges were perfused with different flow rates adjusted between 0 to 300 mL/min, corresponding to flow ratios comprised between 300:0 and 150:150. The contrast agent (Levovist, Schering) was injected at constant rates (0.6 to 5 g/h). Sequential pairs of images showing simultaneously the cross-sections of the two filters were acquired with a HDI 5000 (ATL) and the Doppler data were processed with HDI lab software (ATL). The absolute values of the signal in the different regions-of-interest (ROI) were not closely related to flow rate. At the opposite, the rapid signal decrease between the first and the second image of each pair was inversely proportional to the flow rate. An index of perfusion [PerI = image 1/(image 1 -- image 2)] was defined. It correlated closely with the absolute and relative flow rates. For the latter, the slopes of regression were found to be independent of the infusion rate of Levovist. Thus, the use of pairs of images combined with a continuous infusion of Levovist provide a quantification of perfusion. PMID- 11397531 TI - Impaired tensile strength after shock-wave application in an animal model of tendon calcification. AB - Extracorporeal shock-wave application facilitates dissolution of rotator cuff calcifications. Therefore, disappearance or disintegration of tendon calcifications by shock waves might be appropriate for any kind of tendon calcification. Here, shock waves with various energy flux densities were applied to the mineralized medial gastrocnemius tendon of turkeys as an animal model. After application of shock waves in vivo, with energy flux density of 0.6 mJ/mm(2), histologic examination and microradiography did not show dissolution or disintegration of tendon calcifications. After shock-wave application in vitro, even for energy flux density of 1.2 mJ/mm(2) neither dissolution nor disintegration of tendon calcifications were observed. Biomechanical testing revealed significant impairment of tensile strength following shock-wave application in vitro, with energy flux density of 1.2 mJ/mm(2), but not with 0.6 mJ/mm(2). These results are important for considerations of clinical extracorporeal shock-wave application on tendon calcifications, as well as on tendon ossifications. PMID- 11397532 TI - Changes in ultrasound properties of porcine kidney tissue during heating. AB - Changes in the ultrasound (US) attenuation and backscatter of fresh pig kidney were measured as the tissue was heated. The objective was to use these changes to predict how an US image would change in real-time with a view to its use as a monitoring tool for minimally invasive thermal therapy (MITT). Separate samples of fresh pig kidney were heated from 37 degrees C to temperatures of 45 degrees, 50 degrees, 55 degrees, 60 degrees and 65 degrees with warm water. Measurements were made over the frequency range from 3.5 MHz to 7.0 MHz during 30-min heating experiments. A general increase in attenuation magnitude (dB/cm) and slope (dB/cm MHz) was observed at temperatures of 55 degrees C or greater. Little change in backscatter power was observed during heating to 45 degrees C. At higher temperatures, the changes in backscatter showed a more complex pattern throughout the experiments, but still showed a trend of increase to a greater value at the end of heating than at the start. This backscatter increase was greater at higher temperatures. The net effect of the changes in US properties suggests that it may be possible to use diagnostic US to monitor, in real-time, MITT in kidney. PMID- 11397533 TI - The mechanisms of stone fragmentation in ESWL. AB - Currently, several mechanisms of kidney stone fragmentation in extracorporal shockwave lithotripsy (ESWL) are under discussion. As a new mechanism, the circumferential quasistatic compression or "squeezing" by evanescent waves in the stone has been introduced. In fragmentation experiments with self-focussing electromagnetic shock-wave generators with focal diameters comparable to or larger than the stone diameter, we observed first cleavage surfaces either parallel or perpendicular to the wave propagation direction. This is in agreement with the expectation of the "squeezing" mechanism. Because, for positive pulse pressures below 35 MPa and stones with radii of 15 mm or smaller, cleavage into only two fragments was observed, we developed a quantitative model of binary fragmentation by "quasistatic squeezing." This model predicts the ratio of the number of pulses for the fragmentation to 2-mm size and of the number of pulses required for the first cleavage into two parts. This "fragmentation-ratio" depends linearly alone on the stone radius and on the final size of the fragments. The experimental results for spherical artificial stones of 5 mm, 12 mm and 15 mm diameter at a pulse pressure of 11 MPa are in good agreement with the theoretical prediction. Thus, binary fragmentation by quasistatic squeezing in ESWL as a new efficient fragmentation mechanism is also quantitatively verified. PMID- 11397534 TI - Use of overpressure to assess the role of bubbles in focused ultrasound lesion shape in vitro. AB - Overpressure--elevated hydrostatic pressure--was used to assess the role of gas or vapor bubbles in distorting the shape and position of a high-intensity focused ultrasound (HIFU) lesion in tissue. The shift from a cigar-shaped lesion to a tadpole-shaped lesion can mean that the wrong area is treated. Overpressure minimizes bubbles and bubble activity by dissolving gas bubbles, restricting bubble oscillation and raising the boiling temperature. Therefore, comparison with and without overpressure is a tool to assess the role of bubbles. Dissolution rates, bubble dynamics and boiling temperatures were determined as functions of pressure. Experiments were made first in a low-overpressure chamber (0.7 MPa maximum) that permitted imaging by B-mode ultrasound (US). Pieces of excised beef liver (8 cm thick) were treated in the chamber with 3.5 MHz for 1 to 7 s (50% duty cycle). In situ intensities (I(SP)) were 600 to 3000 W/cm(2). B mode US imaging detected a hyperechoic region at the HIFU treatment site. The dissipation of this hyperechoic region following HIFU cessation corresponded well with calculated bubble dissolution rates; thus, suggesting that bubbles were present. Lesion shape was then tested in a high-pressure chamber. Intensities were 1300 and 1750 W/cm(2) ( +/- 20%) at 1 MHz for 30 s. Hydrostatic pressures were 0.1 or 5.6 MPa. At 1300 W/cm(2), lesions were cigar-shaped, and no difference was observed between lesions formed with or without overpressure. At 1750 W/cm(2), lesions formed with no overpressure were tadpole-shaped, but lesions formed with high overpressure (5.6 MPa) remained cigar-shaped. Data support the hypothesis that bubbles contribute to the lesion distortion. PMID- 11397535 TI - A comparison of the hemolytic potential of Optison and Albunex in whole human blood in vitro: acoustic pressure, ultrasound frequency, donor and passive cavitation detection considerations. AB - This project tested the hypothesis that a "second-generation" ultrasound (US) contrast agent (Optison), offering extended echogenicity over that of its "first generation" predecessor (Albunex), would have the greater potential for sonolysis of human erythrocytes in vitro. Whole human blood, obtained from apparently healthy donors, was anticoagulated and subsequently exposed in vitro to US in the presence of one of each or neither of the two US contrast agents. The US exposures were for 30 s and involved frequency (1.0, 2.2 and 3.4 MHz) and amplitude (approximately 2.8 to 0.38 MPa P(-)) regimens; pulse duration (200 micros) and interpulse interval (20 ms) were held constant. The data supported the hypothesis, with an overall ratio of approximately 2.5 for relative extent of background-corrected US-induced hemolysis of the Optison/Albunex regimens. Passive cavitation detection analyses corroborated the results obtained with hemolysis. PMID- 11397536 TI - Doppler sonography measurement of portal blood flow velocity after glucagon injection in patients with chronic HCV infection. AB - Doppler sonography measurement of portal flow velocity (PFV) after glucagon injection was performed in 45 patients with chronic hepatitis C virus (HCV) infection. Patients were divided into three groups: group 1 = no or mild liver fibrosis; group 2 = moderate to severe liver fibrosis, and group 3 = liver cirrhosis. All patients were examined using a Doppler ultrasound (US) multipurpose equipment and a convex 3.5-MHz probe, 10 min before (baseline), as well as 5 and 10 min after, IV administration of 1 mg of glucagon chloride. No significant differences were found in mean baseline PFV among group 1 (19.4 +/- 2.4 cm/s), group 2 (20.1 +/- 3.6 cm/s) and group 3 (17.5 +/- 3.7 cm/s). Five minutes after glucagon injection, all three groups showed significantly increased values of mean PFV (25.6 +/- 4.8, 23.7 +/- 4.0 and 19.5 +/- 5.0 cm/s, respectively; p < 0.05 vs. baseline). The mean increase of PFV above baseline was significantly higher in group 1 (7.9 +/- 3.7 cm/s) than in group 2 (4.5 +/- 3.9 cm/s) (p < 0.05) or in group 3 (2.7 +/- 2.3 cm/s) (p < 0.05). A significant inverse correlation was found between individual values of fibrosis score and of individual increase of PFV. In patients with chronic HCV infection, Doppler sonography measurement of PFV after glucagon injection could be useful in assessing the severity of liver histological damage. PMID- 11397537 TI - On the fine structure of the pes Hippocampi major (with plates XIII-XXIII). 1886. AB - We have provided a translation of Golgi's original paper on the mammalian hippocampus (first published in 1883 and reprinted numerous times), along with a preface on its historical context. Golgi believed that this part of the cerebral hemisphere showed best the exact relationship between nerve cells and nerve fibers, the most important problem in 19th century neuroscience. PMID- 11397538 TI - Connections of the auditory cortex with the claustrum and the endopiriform nucleus in the cat. AB - We studied the connections of eleven auditory cortical areas with the claustrum and the endopiriform nucleus in the cat, by means of cortical injections of either wheat germ agglutinin conjugated to horseradish peroxidase, or biotinylated dextran amines. Unlike previously accepted reports, all auditory areas have reciprocal connections with the ipsi- and contralateral claustrum, though they differ in strength and/or topography. The areas that send the strongest projections are the intermediate region of the posterior ectosylvian gyrus and the insular cortex, followed by the primary auditory cortex and the dorsal portion of the posterior ectosylvian gyrus. The high degree of convergence of cortical axons in the intermediate region of the claustrum, arising from tonotopic and nontonotopic areas, suggests that claustral neurons are unlikely to be well tuned to the frequency of the acoustic stimulus. Corticoclaustral axons from any given area cover territories largely overlapping with those occupied by the claustrocortical neurons projecting back to the same area. The location of cortically projecting neurons in the claustrum matches the position of the target cortical area in the cerebral hemisphere, both rostrocaudally and dorsoventrally. These findings suggest that the intermediate region of the claustrum integrates inputs from all auditory cortical areas, and then sends the result of such processing back to every auditory cortical field. On the other hand, the endopiriform nucleus, a limbic-related structure thought to play a role in the acquisition of conditioned fear, would process mostly polymodal information, since it only receives projections from the insular and temporal cortices. PMID- 11397539 TI - Mu opioid receptor efficacy and potency of morphine-6-glucuronide in neonatal guinea pig brainstem membranes: comparison with transfected CHO cells. AB - The major side effect of morphine and its active metabolite, morphine-6 glucuronide (M6G), is respiratory depression, which is mediated by mu opioid receptors in the medulla and pons. Although the effect of morphine on coupling between mu opioid receptors and G proteins has been studied, the effect of M6G on this coupling has not. Therefore, stimulation of guanylyl-5'-O-([gamma(35)S] thio)-triphosphate ([(35)S]-GTPgammaS) binding by these two narcotic analgesic drugs was compared to the mu-specific synthetic opioid peptide [D-Ala(2), N MePhe(4), Gly-ol(5)]enkephalin in Chinese hamster ovarian cells stably transfected with the murine mu opioid receptor and in brainstem membranes prepared from 3-, 7-, and 14-day-old guinea pigs. All three agonists stimulated [(35)S]-GTPgammaS binding in transfected cells and neural tissue, and the stimulation was antagonized by naloxone. In brainstem membranes, but not transfected cells, M6G was less efficacious but more potent than morphine, which may be due to differences between murine and guinea pig mu opioid receptors or in the G proteins in these two tissues. Efficacy of the agonists did not change during development, but overall potency decreased between 3 and 14 days after birth. In vivo potency differences for respiratory depression between morphine and M6G are qualitatively similar to in vitro potency differences of these drugs to stimulate [(35)S]-GTPgammaS binding in neonatal guinea pig brainstem membranes. Tolerance to opioid effects on [(35)S]-GTPgammaS binding developed in transfected cells incubated with morphine with the maximum decrease in potency occurring 18 h later than the maximum decline in efficacy. PMID- 11397540 TI - Omega-conotoxin GVIA potently inhibits the currents mediated by P2X receptors in rat DRG neurons. AB - We examined effects of omega-conotoxin previously known as a selective blocker of N-type calcium channels, on the adenosine triphosphate (ATP)-induced currents in the rat dorsal root ganglion neurons. These neurons express at least two types of ionotropic purinoreceptors: P2X3 receptors that have very rapid desensitization kinetics and P2X2/X3 heterooligomeric receptor, which exhibits slow desensitization. We have found that omega-conotoxin GVIA potently inhibits the inward currents mediated by both receptor types. This effect was specific for the receptor subtypes: the IC(50) value for responses evoked by 10 microM ATP was 21.2 +/- 1.7 nM for the P2X3 receptor-mediated responses and 3.84 +/- 0.43 microM for slower responses mediated by P2X2/X3 heteropolymers. The efficacy of another type of omega-conotoxin, MVIIC, is much lower: at 10 microM the latter toxin inhibited the rapidly desensitizing response by 65% and the slowly desensitizing response by 18%. The effects of both toxins were reversible and independent on the membrane potential. Omega-Conotoxin GVIA shifted the dose dependence for the agonistic action of ATP on P2X3 receptors to higher concentrations without producing any effect on the kinetics of the response. It is suggested that omega conotoxin allosterically modulates the receptor properties, rather than competes for the agonist binding site. PMID- 11397541 TI - Alterations in signal transduction cascade in young and adult rat brain and lymphocytes. AB - Signal transduction cascade, phosphoinositide metabolism, and protein kinases were studied from discrete areas of rat brain like cerebral hemispheres, cerebellum, brainstem, and diencephalon as well as lymphocytes isolated from three different age groups of rats; young (1 month), young adult (3-4 months), and adult (12 months) rats. The activities of protein kinase A, protein kinase C, phospholipase A(2) and phospholipase C and inositol 1, 4, 5-triphosphate, diacylglycerol, cyclic adenosine monophosphate contents were assayed from different brain areas and lymphocytes from these three age group rats. An upregulatory effect on the signal transduction system was observed from 1 month to 3-4-month age group, whereas, the brain tissue and lymphocytes of adult rats showed lower contents and activities of signal transduction components as compared to young adults. In view of the established 'cross talk' between signal transduction system, the present results suggests that molecular/cellular changes in brain and immune cells signal transduction pathway along with neuronal cell loss may contribute to age-related decline in nervous as well as immune system functions. PMID- 11397542 TI - Further evidence for a significant participation of the melanocortin 4 receptor in the preovulatory prolactin surge in the rat. AB - We previously reported that the melanocortin 4 receptor may play a significant role in mediating the preovulatory surges of luteinizing hormone and prolactin in the rat. In order to confirm this previous finding, in the present study we examined and compared the effects of intracerebroventricular administrations of 1.0 nmol of MT II (a non-selective melanocortin 3 and 4 receptor agonist) and 10 nmol of gamma(1)-melanocyte-stimulating hormone (a selective melanocortin 3 receptor agonist) on luteinizing hormone and prolactin surges in starved, gonadal steroid-primed ovariectomized female rats, which is a model deprived of inherent surges of the two hormones. MT II significantly recovered the surge of prolactin, but not of luteinizing hormone (although a tendency to increase was seen), and gamma(1)-melanocyte-stimulating hormone was without effect on both hormones. This study corroborated our previous report through a different and direct approach that the melanocortin 4 receptor, but not the melanocortin 3 receptor, plays a significant role in mediating the preovulatory prolactin surge in the rat. PMID- 11397543 TI - Effects of morphine on pentobarbital-induced responses in mu-opioid receptor knockout mice. AB - Effects of morphine on the potentiation of pentobarbital-induced responses were investigated using mu-opioid receptor knockout mice. The duration of loss of righting reflex, hypothermia, and loss of motor coordination induced by pentobarbital were measured after pretreatment with either morphine or saline. Morphine pretreatment failed to show potentiation of both pentobarbital-induced loss of righting reflex and hypothermia in mu-opioid receptor knockout mice, while it significantly potentiated these responses in the wild-type controls. For motor incoordination test, morphine potentiated pentobarbital-induced motor incoordination in the wild-type mice. However, morphine may have opposite effects in the mu-opioid receptor knockout mice. These results demonstrate that synergism between morphine and pentobarbital is not detected in mu-opioid receptor knockout mice and that potentiation of pentobarbital-induced loss of righting reflex and hypothermia by morphine is mediated through mu-opioid receptor. It was interesting to note that pentobarbital-induced decrease in body temperature was less severe in mu-opioid receptor knockout mice than in wild-type mice. PMID- 11397544 TI - Spider neurotoxins block the beta scorpion toxin-induced calcium uptake in rat brain cortical synaptosomes. AB - In this paper we describe the effects of the beta scorpion toxin Tityus gamma (TiTX gamma) and spider neurotoxins Tx3-3 and Tx3-4 in the (45)Ca(2+) uptake in synaptosomes. The TiTX gamma-stimulatory effect on (45)Ca(2+) uptake in synaptosomes was inhibited omega-Conotoxin MVIIC (omega-CgTX MVIIC) (0.1 microM) and omega-Agatoxin IVA (0.1 microM) by 70% and 41%, respectively. omega-CgTX MVIIC (1.0 microM) almost completely blocked the TiTX gamma-induced (45)Ca(2+) uptake in synaptosomes. Verapamil (1.0 microM) and omega-Conotoxin GVIA (0.1 microM) had no effect in the scorpion toxin-induced (45)Ca(2+) influx. The spider neurotoxins Tx3-3 and Tx3-4 inhibited the TiTX gamma-induced calcium uptake with an IC(50) of 10.0 and 30.0 nM, respectively. It is suggested that spider neurotoxins Tx3-3 and Tx3-4 blocking effect in the TiTX gamma-induced calcium uptake involves P/Q-type calcium channels. PMID- 11397545 TI - Differential roles of spinal cholera toxin- and pertussis toxin-sensitive G proteins in nociceptive responses caused by formalin, capsaicin, and substance P in mice. AB - The aim of the present study is to characterize the roles of spinal cholera toxin (CTX)- and pertussis toxin (PTX)-sensitive G proteins in the regulation of various nociceptive responses. The effects of intrathecal (i.t.) pretreatments with CTX and PTX on the formalin (subcutaneous)-, capsaicin (i.t.)-, and substance P (SP; i.t.)-induced nociceptive behaviours were examined in mice. Pretreatment with CTX (i.t.; 24 h before) significantly and dose-dependently (0.05-0.5 microg) suppressed both the first and second phases of the formalin induced nociceptive behaviour. On the other hand, pretreatment with PTX (i.t., 6 days before) at the same doses (0.05-N0.5 microg) did not affect the formalin induced response. Capsaicin (i.t., 0.5 microg)- and SP (i.t., 0.7 microg)-induced nociceptive behaviours were attenuated by the pretreatment with CTX. In addition, SP-induced nociceptive response was also attenuated by the pretreatment with PTX. However, the capsaicin-induced nociceptive response was not influenced by PTX pretreatment. These findings suggest that, at the spinal cord level, CTX sensitive G-proteins are involved in the formalin-, capsaicin-, and SP-induced nociceptive behavioural responses, whereas PTX-sensitive G proteins are involved in SP-induced nociceptive response. PMID- 11397546 TI - Tuning to Y-like figures in the cat striate neurons. AB - Sensitivity to symmetric or asymmetric Y-like figures and crosses of different shapes and orientations flashed in the receptive field was studied in 101 neurons of the cat striate cortex (area 17) and compared with their orientation tuning to a single light bar. Selective sensitivity to the Y-like figure (figure/bar response ratio more than 1.25) was found in 78/101 neurons (77.2% of cases) and to the cross-in 54/101 units (53.4%). In 62.5% of neurons with sensitivity to both figures, sensitivity to the Y-like figure was higher than to a cross. Tuning to Y-like figure was typically (60%) selective to both its shape and orientation. The remaining Y-like selective neurons exhibited invariant tuning to orientation and/or shape of the figure. The preferred angles between two lines of Y-like figures were distributed in the range of 22.5-157.5 degrees with slight preference to 90 degrees, while crosses of 45 degrees and 90 degrees angles were preferable. Response magnitudes to a single bar, a Y-like figure and a cross were positively correlated. Possible mechanisms and functional implication of the striate sensitivity to Y-like figures are discussed. PMID- 11397547 TI - alpha(2C)-Adrenoceptors modulate the effect of methylphenidate on response rate and discrimination accuracy in an operant test. AB - The present study investigated the role of alpha(2C)-adrenoceptors in the regulation of activity and discrimination accuracy in an operant chamber test. We trained food deprived control and alpha(2C)-adrenoceptor knockout mice to collect liquid food rewards in an operant chamber during the light (20 s) period. No food reward was delivered during the dark period (40 s). The alpha(2C)-adrenoceptor knockout mice tended to make fewer total responses and collect less rewards than their controls after saline treatment. However, only response accuracy of alpha(2C)-adrenoceptor knockout mice was significantly lower than that of the control mice. Methylphenidate, a drug blocking dopamine re-uptake and increasing dopamine release, dose-dependently decreased the number of total responses and collected food rewards in control mice but increased those measures in alpha(2C) adrenoceptor knockout mice. In addition, the effect of methylphenidate on discrimination accuracy differed between knockout and control mice. Our results indicate that alpha(2C)-adrenoceptors may regulate dopamine-mediated functions. PMID- 11397548 TI - New spatial cognition tests for mice: passive place avoidance on stable and active place avoidance on rotating arenas. AB - Dry arenas are a convenient tool for assessing the spatial navigation abilities of rodents. In this paper, mice must avoid a punished sector of a dry arena from which they are expelled by a puff of compressed air. The position of the punished sector is defined relative to the coordinate system of the room. In a stable environment the mice can use both extramaze and intramaze landmarks to orient themselves accurately. However, when the shock area is defined by extramaze landmarks, continuous rotation of the arena at 1 rpm makes it impossible to solve the avoidance task using arena-based cues or idiothesis. The avoidance can only be solved by paying attention to extramaze cues. Our protocol tested spatial abilities on stable and rotating arenas. The acquisition of the task was manifested under both conditions by a significant improvement of performance within the first session (short-term memory component) and at the beginning of the 24-h delayed second session (long-term memory component). PMID- 11397549 TI - Sign of lipid peroxidation as measured in the urine of patients with probable Alzheimer's disease. AB - Free radical-induced oxidative damage may be involved in the neurodegenerative process associated with Alzheimer's disease (AD). 8-Isoprostaglandin F(2alpha) (iPF(2alpha)-III) is an isoprostane derived from free radical-induced non enzymatic oxidation of arachidonic acid. It is formed in vivo and is an indicator of lipid peroxidation. Measurements were made of iPF(2alpha)-III in the urine of patients with mild to moderate dementia associated with probable AD and compared to those in the urine of non-demented subjects, who were similar in age and gender. 2,3-Dinor thromboxane B(2) (dinor TXB(2)), a urinary metabolite of TXB(2) was also measured, and served as an indicator of the enzymatic transformation of a product of arachidonic acid. Enzyme linked immunoassays were used to measure iPF(2alpha)-III and dinor TXB(2) in the urine. The concentration of iPF(2alpha) III was significantly elevated in urine of patients assessed to have mild to moderate dementia as compared to non-demented patients. The concentration of urinary dinor TXB(2) was also significantly elevated in the patients with dementia and probable AD as compared to the non-demented subjects. There was considerable overlap of values obtained for demented and non-demented patients for iPF(2alpha)-III and dinor TXB(2), respectively. The observed elevation of iPF(2alpha)-III suggests that patients with mild to moderate dementia associated with probable AD are experiencing significant oxidative stress. This finding is consistent with current data suggesting that oxidative stress may be occurring in patients with dementia and probable AD. The increase of dinor TXB(2) may indicate that enzymatic processes related to the metabolism of arachidonic acid-derived products are also increased in demented patients with probable AD. PMID- 11397550 TI - The development of NADPH-diaphorase and nitric oxide synthase in the visual system of the cichlid fish, Tilapia mariae. AB - The pattern of NADPH-diaphorase expression was studied in the retina and optic tectum of the cichlid fish Tilapia mariae during the first developmental stages. NADPH-diaphorase activity was seen early, at hatching. In the retina a few cell bodies of the retinal inner nuclear layer showed a faint labeling. Scattered labeled cells were found in the stratum periventriculare of the optic tectum, while the optic nerve was unlabeled. Two days after hatching, the number of labeled neurons increased in the inner nuclear layer and a few stained cell bodies were also scattered in the ganglion cell layer. Both the inner and outer plexiform layers showed a diffuse staining and the optic nerve was devoid of labeling. In the optic tectum several positive cells in the periventricular layer, with their dendritic trees extending in the superficial fibrous layer, were found. In 1-month-old Tilapia, NADPH-diaphorase staining and nitric oxide synthase immunoreactivity were found to overlap in both the retina and optic tectum. The density of NADPH-diaphorase labeled neurons in the inner nuclear layer of the retina and in the stratum periventriculare of the optic tectum was largely reduced in comparison with 2 days posthatching embryos. These findings indicated an early and transient production of nitric oxide in the retina and optic tectum of Tilapia, suggesting a functional role for nitric oxide in the development of visual structures in aquatic vertebrates. PMID- 11397551 TI - Three-dimensional estimation of the distribution and size of putative functional units in rat gustatory cortex as assessed from the inter-neuronal distance between two neurons with correlative activity. AB - Two neurons recorded simultaneously in rat gustatory cortex often show, when the oral mucosa is stimulated with effective taste solutions, correlative activities (CAs), which can be quantified with a cross-correlation technique. It has been suggested that the CAs are involved in taste quality discrimination and the neuron pairs with CA (CA pairs) may function as a unit (putative functional unit). The present experiment was undertaken (1) to determine the three dimensional size of putative functional units in rat gustatory cortex on the basis of the interneuronal distance (IND) in individual CA pairs, and (2) to examine to what extent the CA is related to the intrapair similarity in taste responses. The findings were as follows: (1) The CAs occurred in 30 out of 55 pairs with INDs < or =270 microm and often at INDs <100 microm. (2) The numbers of putative functional units were calculated tentatively by using the longest or the mean INDs; at least 200 or over 4,000 units, respectively. (3) In the CA pairs, the "best taste" was identical (homo-type) in many pairs, the "taste profiles" were often similar, and the temporal courses of taste responses resembled each other in more than one half of the pairs. A possible role of hetero-type pairs was discussed. (4) In a small number of cases, troughs were observed in the CCs, suggesting the involvement of local inhibitory neurons. In conclusion we point out that nearby neurons are prone to CA, if (1) their best tastes are identical, (2) their taste profiles are similar, and (3) the temporal patterns of taste response resemble each other. PMID- 11397552 TI - Cytotoxicity of potential ocular permeation enhancers evaluated on rabbit and human corneal epithelial cell lines. AB - A series of prospective ocular permeation enhancers, benzalkonium chloride (BAC), cetylpyridinium chloride (CPC), ethylenediaminetetraacetic acid (EDTA), polyoxyethylene (20) stearyl ether (PSE) and polyethoxylated castor oil (PCO) were tested for cytotoxicity on cultures of rabbit (RCE) and human (HCE) corneal epithelial cells. The cells were treated for 5,15 and 60 min with different concentrations of the test substances, in serum-free medium and in medium containing 15% foetal bovine serum (FBS). The cytotoxicity was evaluated by WST-1 test. The EC(50) values for HCE, after 15 min exposure and in the presence of FBS, indicate the following order of cytotoxicity: PSE> or =BAC>CPC>EDTA>PCO. After 1 h exposure the order of decreasing cytotoxicity was PSE> or =BAC>CPC>PCO>EDTA. In all cases the presence of FBS appeared to exert a protective effect against the cytotoxic effect. PMID- 11397553 TI - Modulation of arachidonic acid turnover in macrophages by cadmium. AB - The effects of cadmium (Cd) induced redox changes on arachidonic acid (AA) turnover in mouse resident peritoneal macrophages (pM) were studied. The pre incubation of pM in a medium containing glutathione (GSH, 0.1 or 1 mM) for 6 h protects pM from loss of viability and AA uptake diminution induced by Cd with regard to non pre-incubated cultures. The exposure of macrophages to Cd 10 microM decreases AA uptake within 2 h and increases AA release in relation to non exposed macrophages. It also enhances AA mobilization and reactive oxygen species (ROS) release induced by okadaic acid and opsonized zimosan and decreases those induced by lipopolysaccharide, but does not modify either AA mobilization or ROS release induced by phorbol ester. These results might suggest that redox changes induced by Cd produce an important impact on AA turnover in macrophages; information that is relevant in the understanding of the cellular toxicity of this metal. PMID- 11397554 TI - Microcystin-induced down-regulation of lymphocyte functions through reduced IL-2 mRNA stability. AB - Here we report that lymphocyte functions were down-regulated by cyanobacterial hepatotoxin microcystin. Treatment of three microcystin (MC) isotypes, MC-LR, MC YR and nodularin, on B6C3F1 mouse splenocytes produced dose-dependent inhibition of in vitro polyclonal antibody response and lymphoproliferation to LPS. ConA induced lymphoproliferative response was decreased by MC-YR and nodularin, but no significant effect was observed in the MC-LR treatment. Intraperitoneal administration of nodularin into B6C3F1 mice decreased humoral immune responses to sheep red blood cell (sRBC), and the inhibitory effect became severe when hepatic uptake of nodularin was blocked by rifampicin. Each MC 1 microM suppressed phorbol 12-myristate 13-acetate (PMA) plus ionomycin-induced IL-2 mRNA expression in splenocytes and thymocytes, but not in EL-4 mouse thymoma cells. To further characterize the mechanism for the reduced IL-2 mRNA level, IL-2 mRNA stability was measured using RT-PCR. Deprivation of PMA/ionomycin stimuli from activated splenocytes and blockade of new transcription resulted in destabilization of IL-2 mRNA, which was accelerated by MC treatment. These results demonstrated that MC down-regulated lymphocyte functions and the immunosuppression was mediated, at least in part, through decreased IL-2 mRNA stability. PMID- 11397555 TI - Topical and oral administration of the natural water-soluble antioxidant from spinach reduces the multiplicity of papillomas in the Tg.AC mouse model. AB - The Tg.AC mouse carrying the v-Ha-ras structural gene is a useful model for the study of chemical carcinogens, especially those acting via non-genotoxic mechanisms. This study evaluated the efficacy of the non-toxic, water-soluble antioxidant from spinach, natural antioxidant (NAO), in reducing skin papilloma induction in female hemizygous Tg.AC mice treated dermally five times over 2.5 weeks with 2.5 microg 12-O-tetradecanoylphorbol-13-acetate (TPA). The TPA-only group was considered as a control; the other two groups received, additionally, NAO topically (2 mg) or orally (100 mg/kg), 5 days/week for 5 weeks. Papilloma counts made macroscopically during the clinical observations showed a significant decrease in multiplicity (P<0.01) in the NAO topically treated group. According to histological criteria, papilloma multiplicity were lower in both topical-NAO and oral-NAO groups, but significantly so only in the oral-NAO mice (P<0.01). The beneficial effect of NAO in the Tg.AC mouse is reported. PMID- 11397556 TI - Effect of diallyl sulfide and diallyl disulfide, the active principles of garlic, on the aflatoxin B(1)-induced DNA damage in primary rat hepatocytes. AB - The objective of this study was to investigate the effect of the active principles in garlic-- diallyl sulfide (DAS) and diallyl disulfide (DADS)--on aflatoxin B(1) (AFB(1))-induced DNA damage in primary rat hepatocytes. Primary rat hepatocytes, induced with DNA damage using 10 microM AFB(1) were used as an experimental model. According to the results of LDH leakage, 0.5 and 2 mM of DAS or 0.5 and 1 mM of DADS significantly increased the viability of hepatocytes compared with the AFB(1) controls after 4, 8 and 24 h treatment (P<0.05). According to the results of unscheduled DNA synthesis (UDS) test, 0.5 and 2 mM of DAS or 0.5 and 1 mM of DADS could significantly decrease the DNA damage induced by AFB(1) (P<0.05). Furthermore, 0.5 and 2 mM DAS or 0.5 and 1 mM DADS could increase the glutathione S-transferase (GST) and glutathione peroxidase (GPx) activities as compared with the AFB(1) controls after 24 h treatment (P<0.05). Results of immunoblot analysis of cytosolic GST isoenzyme indicate that the levels of GST isoform Ya, Yb2 and Yc were markedly increased after treatment with 0.5 and 2 mM DAS or 0.5 and 1 mM DADS compared with the AFB(1) control. These results indicate that 0.5 and 2 mM DAS or 0.5 and 1 mM DADS might protect hepatocytes from AFB(1)-induced DNA damage via increasing the activities of GST and GPx. PMID- 11397557 TI - 2-Methoxyethanol metabolism, embryonic distribution, and macromolecular adduct formation in the rat: the effect of radiofrequency radiation-induced hyperthermia. AB - Exposure of pregnant rats to the solvent 2-methoxyethanol (2ME) and radiofrequency (RF) radiation results in greater than additive fetal malformations (Nelson, B.K., Conover, D.L., Brightwell, W.S., Shaw, P.B., Werren, D.W., Edwards, R.M., Lary, J.M., 1991. Marked increase in the teratogenicity of the combined administration of the industrial solvent 2-methoxyethanol and radiofrequency radiation in rats. Teratology 43, 621-34; Nelson, B.K., Conover, D.L., Shaw, P.B., Werren, D.W., Edwards, R.M., Hoberman, A.M., 1994. Interactive developmental toxicity of radiofrequency radiation and 2-methoxyethanol in rats. Teratology 50, 275-93). The current study evaluated the metabolism of 14C-labeled 2ME and the distribution of methoxyacetic acid (MAA) in maternal and embryonic tissues of pregnant Sprague-Dawley rats either exposed to 10 MHz RF radiation or sham conditions. Additionally, adduct formation for both plasma and embryonic protein was tested as a possible biomarker for the observed 2ME/RF teratogenicity. Rats were administered [ethanol-1,2-(14)C]-2ME (150 mg/kg, 161 microCi/rat average) by gavage on gestation day 13 immediately before RF radiation sufficient to elevate body temperature to 42 degrees C for 30 min. Concurrent sham- and RF-exposed rats were sacrificed at 3, 6, 24 or 48 h for harvest of maternal blood, urine, embryos and extra-embryonic fluid. Tissues were either digested for determination of radioactivity or deproteinized with TCA and analyzed by HPLC for quantification of 2ME metabolites. Results show the presence of 2ME and seven metabolites, with the major metabolite, MAA, peaking at 6 h in the tissues tested. MAA, the proximal teratogen, was detectable in maternal serum, urine, embryo and extraembryonic fluid 48 h after dosing. Clearance of total body 14C was significantly reduced for the RF-exposed animals (P<0.05) for the 24-48 h period, but MAA values for serum, embryos and extraembryonic fluid were similar for both sham- and RF-exposed rats. Additionally, no difference was noted for 2ME metabolite profiles in urine or tissue for sham- or RF-exposed rats, thus eliminating an effect of RF radiation on MAA production as a possible explanation for the reported RF-2ME synergism. Subsequently, serum and embryo protein-bound adducts were evaluated by analysis of covalently bound radioactivity. Serum protein binding was significantly higher for sham than RF rats at 3- and 6-h - highest for sham rats at 6 h (519+/-95 microg as parent 2ME/g of protein) whereas RF serum values were highest at 24 h (266+/-79 microg/g protein). Embryonic protein binding was significantly higher for sham rats at 6 h, but binding was highest for both groups at 24 h (sham=229+/-71 microg/g, RF=185+/-48 microg/g). Formation of protein adducts after 2ME is thought to be related to levels of methoxyacetaldehyde, a reactive intermediate in the formation of MAA. These results suggest that no direct relationship exists for covalent binding in the embryo which would explain RF-2ME synergistic malformations. In comparison with urinary metabolites, the relatively slow elimination of adducted serum 2ME indicates that analysis of protein-bound concentrations could be a potential tool for long- term biomonitoring of worker exposure. PMID- 11397558 TI - The acute effects of mono(2-ethylhexyl)phthalate (MEHP) on testes of prepubertal Wistar rats. AB - A single oral dose of 400 mg/kg body weight of mono(2-ethylhexyl)phthalate (MEHP), the testis toxic metabolite of di(2-ethylhexyl)phthalate, was given to 28 day-old male Wistar rats and the testis toxic effects were investigated 3,6, and 12 h after exposure. Detachment and sloughing of germ cells were observed, and in the Sertoli cells the cytoplasmatic intermediate filament vimentin collapsed. In the immunohistochemical investigation the androgen receptor distribution was unchanged between the control group and treated groups. The expression of the testosterone-repressed-prostatic-message-2 gene in rat testis increased after 3 h, but returned to control levels after 6 and 12 h. Caspase-3 activity increased 3 and 12 h after MEHP exposure. This increase could not be correlated to an increase in DNA fragmentation or increase in apoptotic numbers of germ cells. In conclusion, the effect of MEHP in testis is apparently not involving the androgen receptor. Vimentin localisation in the Sertoli cells, and increased levels of caspase-3 activity appear to be sensitive and early markers of MEHP testis toxicity. PMID- 11397559 TI - Triptolide induced cytotoxic effects on human promyelocytic leukemia, T cell lymphoma and human hepatocellular carcinoma cell lines. AB - Triptolide, a traditional Chinese medicine, has been reported to be effective in the treatment of auto-immune diseases, and it can also induce anti-neoplastic activity on several human tumor cell lines. This study investigates the cytotoxic function and the functional mechanism of triptolide on tumor cells. Promyelocytic leukemia, (HL-60), T cell lymphoma (Jurkat), and human hepatocelluar carcinoma (SMMC-7721) cells were subjected to triptolide treatment, and cell growth inhibition was examined by XTT cell viability assay. Cell death mechanism (apoptosis) was confirmed through DNA fragmentation and DAPI staining. Triptolide inhibited 50% of cell growth (IC(50)) on HL-60 cells at 7.5 nM, Jurkat cells at 27.5 nM and SMMC cells at 32 nM. Characteristic apoptotic features including internucleosomal DNA fragmentation and chromatin condensation were observed in triptolide treated cells. Data from the study indicates that triptolide could induce apoptosis in human tumor cell lines and it may be applicable as a potential chemotherapeutic agent for cancer treatment. PMID- 11397560 TI - Acetaminophen elicits anti-estrogenic but not estrogenic responses in the immature mouse. AB - Acetaminophen is a widely used analgesic, which has exhibited evidence of estrogenic activity in estrogen-receptor positive human breast cancer cells. However, there is limited in vivo experimentation regarding the estrogenicity of acetaminophen. Therefore, the present study examined the in vivo estrogenic potential of acetaminophen using the immature female mouse model. Specifically, C57Bl/6 female mice were treated with acetaminophen (100, 200 and 250 mg/kg per day, i.p.) for 3 consecutive days. Positive controls received estradiol (10 microg/kg per day, i.p.) for 3 consecutive days. Markers of estrogenic activity examined were uterine weight, uterine peroxidase activity and progesterone receptor (PR) up-regulation. The results demonstrated that, at all three doses, acetaminophen did not significantly increase uterine weight, uterine peroxidase activity or PR levels. However, the co-administration of E(2) and acetaminophen (200 mg/kg per day, 3 days, i.p.) resulted in a decrease in the E(2)-induced upregulation of uterine and hepatic nuclear PR (uterine PR values of 39.7+/-6.6 and 23.7+/-3.4 fmol/mg for E(2)+vehicle and E(2)+acetaminophen, respectively, P<0.05). Additionally, the estradiol-induced increase in uterine peroxidase was decreased 75% by acetaminophen (200 mg/kg per day, 3 days, i.p.). Therefore, in the immature mouse model acetaminophen treatment did not elicit estrogenic activity. However, acetaminophen had a limited ability to antagonize the effects of E(2). PMID- 11397561 TI - Vitamin E protects against iron-hexachlorobenzene induced porphyria and formation of 8-hydroxydeoxyguanosine in the liver of C57BL/10ScSn mice. AB - The effect of vitamin E treatment on total porphyrin content, lipid peroxidation (LOOH) and 8-hydroxydeoxyguanosine (8-OHdG) was studied in the livers of C57BL/10ScSn mice following hexachlorobenzene (HCB) and iron treatment. HCB was administered i.p. (totalling 300 mg/kg) twice, with 1 week interval. Three days after the first HCB injection iron-dextran was given i.p. (500 mg Fe per kg). Vitamin E was administered weekly (20 mg/kg) by s.c. injection. Both total hepatic porphyrin and LOOH levels were significantly (P<0.001) increased in the HCB-iron treated group as compared with the control group. Mice treated additionally with vitamin E had significant (P<0.001) lower levels as compared with the HCB-iron group. Similarly, the levels of 8-OHdG were significantly (P<0.001) increased above controls after HCB-iron treatment and this increase was reduced after co-treatment with vitamin E (P<0.02). The data support the hypothesis that the mechanism of hepatic porphyrinogenicity of HCB with iron overload is an oxidative free radical process. PMID- 11397562 TI - Automobile exhaust particle-induced apoptosis and necrosis in MRC-5 cells. AB - To study the effect of particulate extracts (PE) collected from a heavy traffic road in Lanzhou City, on MRC-5 cell apoptosis, and to explore the toxicity action of PE and its mechanism. Cultured MRC-5 cells were incubated in the extracts of different concentrations. Inhibition of proliferation was measured with a colorimetric 3-[4,5-dimethyl thiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. Morphological assessment of apoptosis was performed with fluorescence microscopy and electronic microscopy. Extracted DNA from the cells was electrophoresed on agarose gel in order to observe DNA fragmentation. The amount of apoptotic cells was measured by flow cytometry. The results indicated that exposure of exponentially growing MRC-5 cells exposed to PE 5-160 microg l(-1) for 24-96 h resulted in dose- and time-dependent reduction of survival of MRC-5 cells. After treatment with PE, markedly morphological changes of MRC-5 cells including "apoptotic bodies", were observed with a fluorescence microscope. Agarose gel electrophoresis of DNA from the cells treated with PE for 48 and 72 h revealed a "ladder" pattern. PE induced apoptosis in low doses but necrosis in high doses. Apoptotic rates were 12.95, 17.40 and 29.80% after treatment with PE 5, 10, and 20 microg l(-1), respectively. A typical sub-diploid apoptosis peak was demonstrated in MRC-5 cells treated with PE. A significant dose-effect response and time-effect correlation could be found between apoptosis rates and PE. All results confirmed that the PE could induce and accelerate apoptosis in low doses but necrosis in high doses. PMID- 11397563 TI - Biomedical applications of collagen. AB - Collagen is regarded as one of the most useful biomaterials. The excellent biocompatibility and safety due to its biological characteristics, such as biodegradability and weak antigenecity, made collagen the primary resource in medical applications. The main applications of collagen as drug delivery systems are collagen shields in ophthalmology, sponges for burns/wounds, mini-pellets and tablets for protein delivery, gel formulation in combination with liposomes for sustained drug delivery, as controlling material for transdermal delivery, and nanoparticles for gene delivery and basic matrices for cell culture systems. It was also used for tissue engineering including skin replacement, bone substitutes, and artificial blood vessels and valves. This article reviews biomedical applications of collagen including the collagen film, which we have developed as a matrix system for evaluation of tissue calcification and for the embedding of a single cell suspension for tumorigenic study. The advantages and disadvantages of each system are also discussed. PMID- 11397564 TI - Topical transfection using plasmid DNA in a water-in-oil nanoemulsion. AB - Expression plasmids encoding chloramphenicol acetyltransferase (CAT) or human interferon-alpha2 cDNA were formulated in water-in-oil nanoemulsions and applied to murine skin. The histological location of transfected cells was assessed by in situ DNA PCR and showed that the deposition of plasmid DNA was primarily in follicular keratinocytes. Transgene expression in the skin was monitored for 24 72 h, following topical application of either single or multiple daily doses by quantitative RT-PCR and ELISA. It was found that transgene expression was optimal at 24 h following topical application of a single dose of water-in-oil nanoemulsion containing plasmid DNA. Dose-response studies using a total dose of 3, 10 or 30 microg of plasmid DNA suggested that topical transfection using nanoemulsions is subject to both threshold and saturation effects. None of the cationic liposome formulations tested as controls mediated transgenic protein expression at levels higher than background values of the ELISAs used to assay transgenic protein. Single and multiple dose experiments using human interferon alpha2 as a transgene indicated that the efficiency of nanoemulsion mediated transfection was most effective in the context of normal versus atrophic hair follicles. In addition, the total amount of human interferon-alpha2 present in skin appeared to accumulate as a consequence of multiple dosing. Histologic evaluation of treated skin showed no overt signs of toxicity or irritation associated with the short-term application of the nanoemulsions. The results suggest that water-in-oil nanoemulsions can be used to facilitate transfection of follicular keratinocytes in vivo. PMID- 11397565 TI - The interaction between wheat germ agglutinin and other plant lectins with prostate cancer cells Du-145. AB - The bioadhesive properties of fluorescein-labeled plant lectins with different carbohydrate specificities were investigated by flow cytometry at 4 and 37 degrees C using Du-145 prostate cancer cells. At both temperatures the lectin association rate increased following the order: Dolichos biflorus agglutinin (DBA)or= 60 years) subjects were enrolled in this study. The study involved medical records abstracting and patient interviews. Pain was assessed in 150 (49%) subjects using the question "Do you have pain?" (Group 1). In the remaining 155 (51%) subjects (Group 2), pain was assessed using three pain assessment instruments: the visual analog scale, the behavior (faces) scale, and the pain descriptive scale. The overall frequency of diagnosing pain was greater in the subjects in Group 2 compared to the subjects in Group 1 (30% vs. 15%, P < 0.01). Using the three pain assessment scales greatly increased the frequency of diagnosing pain among the old-old (>or= 85 years) residents (32% Group 2 vs. 6% Group 1, P < 0.001). The frequency of diagnosing pain among cognitively impaired residents showed a similar trend (16% Group 2 vs. 10% Group 1, P = 0.2). These data indicate that using three easily-administered pain assessment scales increased the frequency of diagnosing pain among nursing home residents. PMID- 11397603 TI - Predictors of pain and fatigue in the year following diagnosis among elderly cancer patients. AB - Using data obtained from an inception cohort of 841 patients aged 65 or older newly diagnosed with breast, colon, lung, or prostate cancer, and observed at 6 8, 12-16, 24-30, and 52 weeks, three questions related to patients' experiences with pain and fatigue were posed. First, how do numbers of patients reporting neither pain nor fatigue, either symptom, or both change during the observation year? Second, did number of comorbid conditions, site and stage of cancer, treatment modalities, symptom management medication, and time affect the presence of these two symptoms? Third, do pain and fatigue predict the numbers of co occurring other symptoms? Findings indicate that during the year patients improved with respect to their reports of pain and/or fatigue. Stage, more comorbidity, and lung cancer were related to both pain and fatigue. Chemotherapy was related to reports of fatigue, but did not have an extended effect on fatigue. PMID- 11397604 TI - Sedation for refractory symptoms of terminal cancer patients in Taiwan. AB - This study assessed sedation in terminal cancer patients in terms of three characteristics: frequency; relationship to intractable symptoms; and the extent to which medical staff, family, and patients found sedation to be ethically acceptable and efficacious. Two hundred seventy-six consecutive patients, who were admitted to the palliative care unit of National Taiwan University Hospital in Taiwan between August 1998 and the end of May 1999, were enrolled. A recording form was completed every day. This included demographic data, pain and common symptom scores, and the use of sedation in the terminal phase. Seventy (27.9%) of 251 patients who died received sedation. Sedation was administered to relieve agitated delirium in 40 (57.1%), dyspnea in 16 (22.8%), severe pain in 7 (10%) and insomnia in 5 (7.2%). The drugs used for sedation were haloperidol in 35 (50%), midazolam in 17 (24.3%), and rapidly increasing dosage of morphine in 9 (12.9%). In fewer than half (42.9%) of the patients, sedation was with the consent of both patient and family, and half (50%) had the consent of family alone. The overwhelming majority of medical staff and family felt the decision to use terminal sedation was ethically acceptable. There was no significant difference in survival time between sedated and non-sedated patients (28.49 vs. 24.71 days, t = -0.791, P = 0.430). Positive ethical acceptability and higher satisfaction with symptom control with terminal sedation were found in both medical staff and family in this study. Further work is needed to find the most appropriate time of intervention and to improve management of refractory symptoms in dying patients. PMID- 11397605 TI - Symptom burden at the end of life: hospice providers' perceptions. AB - To describe symptom prevalence, frequency, and severity among hospice patients, from the perspective of hospice providers, a cross-sectional study was conducted among 16 hospices participating in the Population-based Palliative Care Research Network (PoPCRN). Hospice staff estimated symptom presence, frequency, and severity, using the Memorial Symptom Assessment Scale. Among the 348 patients (median age 78 years, 55% female, 55% cancer diagnosis), the most common symptoms noted by hospice staff were lack of energy (83%), pain (76%), lack of appetite (63%), drowsiness (61%), difficulty concentrating (60%), and sadness (51%). When present, lack of energy was rated by hospice providers as both frequent (75% "frequently" or "almost constantly") and severe (46% "severe" or "very severe"). Symptoms varied by care setting and by diagnosis. Hospice staff often lacked sufficient information to rate potentially important symptoms, such as problems with sexual interest or activity (63%), change in self image (30%), and worrying (26%). This study identifies a significant burden of unrelieved symptoms among hospice patients, suggesting a need for more widespread institution of symptom management strategies with proven effectiveness and additional investigation into treatment of common symptoms for which few effective treatment strategies are known. PMID- 11397606 TI - Predicting place of death of elderly cancer patients followed by a palliative care unit. AB - To determine the predictors of death outside the palliative care system for elderly cancer patients who were followed by a palliative care unit (PCU), a retrospective review of 199 charts was performed. Comorbidities, current neoplastic disease (location of tumor, months since diagnosis, number of days of PCU follow-up), symptoms, drug use, and social support were recorded. Place of death was not significantly different among sexes. Factors predicting death in the general hospital for men were digestive comorbidities, vomiting, and weakness. For women, disease of the senses was predictive. Although there were no significant differences with respect to place of death between sexes in an older cancer population followed by a PCU, the factors that predicted which patients will die in the hospital for men were generally related to their medical condition, whereas the predictive factors for women were mainly related to functional dependency and social support. Social support had a trend towards predicting the place of death in women. PMID- 11397607 TI - Use of the CAGE questionnaire for screening problem drinking in an out-patient palliative radiotherapy clinic. AB - To determine the positive rate of the CAGE questionnaire in an outpatient palliative radiotherapy clinic and to examine the association between problem drinking, pain control, and analgesic consumption, patients referred for palliative radiotherapy were screened with the CAGE questionnaire and asked to rate their symptom distress using the modified Edmonton Symptom Assessment System (ESAS). The latter instrument uses 11-point numeric scales (0 = best, 10 = worst). Their daily analgesic consumption in oral morphine equivalent was recorded. A total of 128 patients participated in the study. Only 9 patients answered one of the four CAGE questions affirmatively (positive group). All the rest answered negatively (negative group). The mean pain intensity at index site/overall pain was 4.97 +/- 3.31/3.27 +/- 2.76 for the negative group and 6.29 +/- 4.42/2.89 +/- 3.37 for the positive group. The mean total daily oral morphine equivalent for the negative and positive group were 112.35 +/- 233.58 mg and 36.82 +/- 58.85 mg, respectively. There was no significant difference found in other symptoms in the modified ESAS between these two groups. The positive rate of the CAGE in patients with advanced cancer attending an out-patient radiotherapy clinic was only 7%, and analyses were limited by the small sample size of those with a positive CAGE. Whether our observed low positive rate of CAGE represents the true prevalence of problem drinking or the CAGE questionnaire is an insensitive tool for screening problem drinking in an outpatient palliative radiotherapy clinic requires further investigation. We did not find a statistically significant worse pain intensity nor higher analgesic consumption in patients who screened positive for CAGE questionnaire. PMID- 11397608 TI - A comparison of the affective state and quality of life of chemotherapy patients who do and do not develop chemotherapy-induced oral mucositis. AB - The purpose of this longitudinal study was to compare the quality of life and affective state of patients receiving chemotherapy who developed oral mucositis to patients who did not. Outpatients had their mouths assessed at the beginning of their chemotherapy, completed the Multidimensional Quality of Life scale, Cancer version (MQOLS-CA) and the Profile of Mood States (POMS). Patients again completed the MQOLS-CA and POMS if they developed mucositis during their three cycles (monthly), or if they did not and were exiting the study. Seventy-seven outpatients completed the study; 28 patients developed mucositis and 49 did not. The MQOLS-CA total scores for the entire sample decreased significantly over time (F(1,75) = 25.44, P < 0.001), but there was no group by time interaction, i.e., the change in MQOLS-CA total scores did not depend on mucositis status. While the POMS Total Mood Disturbance scores for the entire sample increased significantly over time (F(1,75) = 19.55, P < 0.001), there was a significant group by time interaction (F(1,75)= 4.85, P = 0.03). Patients who developed mucositis had a significant increase in mood disturbance compared to patients who did not. Further, the POMS subscales of depression and anger showed the same pattern of significant increases. In conclusion, the development of mucositis adversely affected the outpatients' affective states, but not their QOL. PMID- 11397609 TI - Fosphenytoin: an intravenous option for the management of acute trigeminal neuralgia crisis. AB - Timely management of trigeminal neuralgia presenting with severe, sustained, crescendo pain can be difficult with oral medications. More rapid pain control often can be achieved using intravenous phenytoin. Fosphenytoin is a phosphate ester prodrug of phenytoin that is significantly better tolerated parenterally than phenytoin in the treatment of epilepsy. Three patients with trigeminal neuralgia refractory to oral medications and presenting with crisis pain were treated urgently with intravenous fosphenytoin. In each case complete relief of pain was achieved for a duration of two days, affording a window of opportunity to modify oral pharmacotherapeutic strategies or to control pain in preparation for invasive neurosurgical intervention. PMID- 11397610 TI - Treatment of reflex sympathetic dystrophy (CRPS type 1): a research synthesis of 21 randomized clinical trials. AB - A blinded meta analysis was performed on randomized clinical trials (RCT) on the medicinal treatment of reflex sympathetic dystrophy (complex regional pain syndrome type I) to assess the methodological quality and quantify the analgesic effect of treatments by calculating individual and summary effect sizes. The internal validity of 21 RCTs was investigated and the quality weighted summary effect size was calculated using a fixed effect model (Glass Delta). The methodological quality ranged from moderate to good (average 46%). Differences were found between the trials in inclusion/exclusion criteria, treatment methods, duration of treatments and trials, and measurement instruments. Statistical analysis was possible for four subgroups; one evaluating the analgesic effects of sympathetic suppressors in general (n = 12), one subgroup concerning the analgesic effects of guanethidine (n = 6), one investigating the analgesic effect of intravenous regional sympathetic blocks (n = 9), and one subgroup (n = 5) evaluating the analgesic effect of calcitonin. Except for the calcitonin subgroup (P = 0.002), the quality-weighted summary effect size of these subgroups were not significant. No significant analgesic effect by sympathetic suppressing agents could be established. Calcitonin seems to provide effective pain relief in reflex sympathetic dystrophy patients. The results of the present study show that weighting methodological quality influences the magnitude of the effect sizes of specific treatment methods. Future studies should control for methodological quality. PMID- 11397611 TI - Use of antimicrobial agents in veterinary medicine and food animal production. AB - Antimicrobial resistance is a growing area of concern in both human and veterinary medicine. This review presents an overview of the use of antimicrobial agents in animals for therapeutic, metaphylactic, prophylactic and growth promotion purposes. In addition, factors favouring resistance development and transfer of resistance genes between different bacteria, as well as transfer of resistant bacteria between different hosts, are described with particular reference to the role of animals as a reservoir of resistance genes or resistant bacterial pathogens which may cause diseases in humans. PMID- 11397612 TI - The role of beta-lactamase producing bacteria and bacterial interference in streptococcal tonsillitis. AB - The causes of penicillin failure in eradicating group A beta-haemolytic streptococcal pharyngo-tonsillitis are described. The mechanisms accounting for the failure include the presence in the tonsils of beta-lactamase producing bacteria and the absence of bacterial interference. The use of antimicrobials that can overcome and modulate these two phenomena and achieve better cure of the infection is described. PMID- 11397613 TI - Classification of oral cephalosporins. A matter for debate. AB - There are many cephalosporins available and various ways of classifying them for clinical use. Oral cephalosporins probably need a classification of their own. This informal discussion was prompted by the appearance of the recommendations of an expert committee of the Paul Ehrlich Gesellschaft. The views of several other commentators are included. There is considerable individual variation in preference for different styles of classification depending on what the classification is for. PMID- 11397615 TI - Susceptibility of Canadian isolates of Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae to oral antimicrobial agents. AB - We measured the susceptibility of Canadian isolates of three respiratory tract pathogens (Haemophilus influenzae, Moraxella catarrhalis and Streptococcus pneumoniae) to several currently approved antimicrobial agents by two different methods. We also measured the susceptibility of isolates to seven fluoroquinolones. Beta-lactamase was produced by 123/566 (21.7%) of H. influenzae isolates compared with 178/200 (89%) of M. catarrhalis isolates. For S. pneumoniae 83/374 (22.2%) isolates were penicillin resistant and of these 2.1% (8/374) showed high level resistance (MIC > or = 2 mg/l). Regardless of methodology, all fluoroquinolones were highly active against H. influenzae (MIC(90) < or = 0.031 mg/l) and M. catarrhalis (MIC(90) < or = 0.064 mg/l) isolates. Susceptibility of H. influenzae to cefuroxime and amoxycillin/clavulanic acid was 99-100% whereas 84-85.5% were susceptible to cefaclor and cefprozil. Azithromycin susceptibility ranged from 82.6 to 99.2% depending on the method. M. catarrhalis isolates were uniformly susceptible to all agents tested except amoxycillin. Cross-resistance in S. pneumoniae to all non-quinolone agents was concurrent with increasing penicillin resistance as shown by increasing MIC90 values. For the fluoroquinolones tested, the rank order of potency based on MIC(90) values was as follows: gemifloxacin (0.031-0.063 mg/l), trovafloxacin (0.125 mg/l), moxifloxacin (0.125-0.25 mg/l), grepafloxacin (0.125-0.25 mg/l), gatifloxacin (0.5 mg/l), levofloxacin (1 mg/l) and ciprofloxacin (2 mg/l). Our study confirms either a high or increasing prevalence of antimicrobial resistant respiratory pathogens in Canada and also compares the new and old fluoroquinolones and their potential role as therapy for community acquired infections. The prevalence of beta-lactamase positive H. influenzae may have decreased from levels reported in previous studies. PMID- 11397614 TI - Comparative antimicrobial activity of ABT-773, a novel ketolide, tested against drug-resistant Gram-positive cocci and Haemophilus influenzae. AB - The antimicrobial activity of ABT-773, a novel ketolide, was tested against 618 Gram-positive strains collected from various surveillance programmes between 1997 and 2000. ABT-773 has potent activity against Streptococcus pneumoniae (MIC90, < or = 0.03-0.12 mg/l), beta-haemolytic streptococci (MIC90, < or = 0.03 mg/l) and viridans group streptococci (MIC90, < or = 0.03 mg/l), including erythromycin resistant strains. In contrast, ABT-773 was less active against erythromycin resistant Staphylococcus aureus (31% susceptible at < or = 0.25 mg/l), coagulase negative staphylococci (41% susceptible) and enterococci (30% susceptible). Haemophilus influenzae (MIC90, 4 mg/l) was less inhibited by the two ketolides tested, and ABT-773 was generally two- to fourfold more potent than telithromycin. The ketolides appear to have potential clinical use against some Gram-positive species resistant to macrolides. PMID- 11397616 TI - Antimicrobial resistance and serotype distribution of Streptococcus pneumoniae isolates from Crete, Greece. AB - Susceptibility to 14 antibiotics was determined for 125 clinical isolates of Streptococcus pneumoniae collected over a 3-year period in Crete, Greece. Twenty three isolates (18.4%) showed intermediate resistance and 15 (12%) high-level resistance to penicillin. Erythromycin, chloramphenicol, tetracycline, trimethoprim-sulphamethoxazole and sparfloxacin resistance rates were 16.8, 10.4, 19.2, 24.8 and 9.6%, respectively. Multiple resistance was observed in 22 strains. Vancomycin and levofloxacin were the most active agents tested. The most prevalent serotype among penicillin-susceptible pneumococci was 14, followed by 9, 7 and 1, while among penicillin-intermediate or -resistant strains serotype 23 was predominant followed by 19 and 9. These results show that as well as a high level of penicillin resistance in this region, some strains are also resistant to other antibiotics and may show multi-drug resistance. PMID- 11397617 TI - Postantibiotic effects and postantibiotic sub-MIC effects of erythromycin, roxithromycin, tilmicosin, and tylosin on Pasteurella multocida. AB - When intermittent dosing is used during treatment, the concentrations of antibiotics fluctuate and subinhibitory concentrations may occur between doses. Postantibiotic effects (PAEs) and postantibiotic subinhibitory effects (PA SMEs) on bacteria may provide additional, valuable information for the rational use of a drug in clinical practice. In this study tilmicosin was the most active antibiotic tested against P. multocida type D with MICs ranging from 4-16 mg/l. Roxithromycin and tilmicosin induced a statistically significantly longer PAE than did tylosin against P. multocida types A and D (P < 0.05). The duration of PAEs and PA SMEs were proportional to the concentrations of drugs used for exposure. The PA SMEs were substantially longer than PAEs on P. multocida. Tilmicosin had a longer PA SME compared with erythromycin, roxithromycin and tylosin for P. multocida. The computerized incubator used in the present study provided an efficient and convenient determination of PAE and PA SME, allowing frequent measurements of the bacterial growth. PMID- 11397618 TI - Nosocomial bloodstream infections in a Turkish university hospital: study of Gram negative bacilli and their sensitivity patterns. AB - Treatment of nosocomial bacteraemia is usually governed by the surveillance results of the particular unit. Such results are especially important when antimicrobial resistance rates are high. Multiresistant isolates including Gram negatives producing extended-spectrum beta-lactamases have been frequently reported in tertiary care units in Turkey. In this study, antimicrobial susceptibilities of Gram-negative blood isolates (n=348) were determined by microbroth dilution tests. The results showed carbapenems (meropenem and imipenem) to be uniformly more potent in vitro than any other drug against the Enterobacteriaceae. Quinolone antibiotics were more active in vitro than aminoglycosides against a range of bacteria. Gram-negative bloodstream isolates were highly resistant to many antimicrobial agents in the hospital. In order to prevent hospital infection and antimicrobial resistance, surveillance of aetiological agents must be performed regularly. PMID- 11397619 TI - Pharmacodynamics of ceftriaxone and cefixime against community-acquired respiratory tract pathogens. AB - Over the last decade or so there has been a growing interest in routes of antimicrobial administration other than by the conventional intravenous route for institutionalized patients and for some outpatients. Both oral (PO) and intramuscular (IM) routes of administration are less costly than giving antimicrobial agents by vein (IV). In addition, fewer complications such as catheter-related sepsis and phlebitis are associated with non-IV routes of administration. Furthermore, a reduced-dosage, reduced-volume IM administration of ceftriaxone may provide a tolerable route of administration and equivalent bactericidal activities compared with higher dose IV ceftriaxone. The purpose of this study was to determine the time that the drug concentration remained in excess of the minimum inhibitory concentration (MIC) (T > MIC) and the duration of bactericidal activities of ceftriaxone one gram administered IV, ceftriaxone 250 mg given IM and cefixime 400 mg given orally against clinical isolates of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis in adult volunteers. Single doses of each agent were administered and serum concentrations were collected over the standard dosing period of 24 h for all study regimens. Ceftriaxone, regardless of route of administration and dose, resulted in bactericidal activities and T > MIC for 100% of the dosing period for S. pneumoniae, H. influenzae, and M. catarrhalis. Cefixime maintained at least 50% T > MIC and bactericidal activity against both isolates each of H. influenzae and M. catarrhalis. Against both isolates of S. pneumoniae, cefixime achieved T > MIC for at least 50% of the dosing period, but did not maintain bactericidal activity. Reduced dose ceftriaxone given IM seems to be a viable alternative to ceftriaxone IV if the pathogen, susceptibility and infection site are known. Based on T > MIC exceeding 50% of the dosing interval, cefixime would be considered an effective alternative to IV therapy against common respiratory tract pathogens. Clinical studies need to be conducted to confirm these findings. PMID- 11397620 TI - Crossover assessment of serum bactericidal activity of grepafloxacin, ofloxacin and clarithromycin against respiratory pathogens after oral administration to healthy volunteers. AB - Serum bactericidal activity was studied in a crossover manner in 10 volunteers, after 2-day administration of grepafloxacin 600 mg qd, ofloxacin 400 mg bid and clarithromycin 500 mg bid. Bactericidal activity against clinical isolates of Streptococcus pneumoniae, Haemophilus influenzae, Moraxella catarrhalis and Klebsiella pneumoniae, was estimated using a standardized microdilution method. Grepafloxacin was highly active against Gram-negative organisms and adequate against pneumococci (mean, 1:13.3). Clarithromycin was very active against both penicillin-susceptible and penicillin-partially-resistant S. pneumoniae (1:102.5) but had poor activity against H. influenzae (1:3.1). Minor adverse effects were commonly associated with grepafloxacin. PMID- 11397622 TI - Suppression of Mannheimia (Pasteurella) haemolytica serovar 1 infection in lambs by intrapulmonary administration of ovine antimicrobial anionic peptide. AB - In this study, the efficacy of ovine antimicrobial anionic peptide (AP) was assessed in a lamb model of acute pneumonia. A single intratracheal dose of the peptide, H-DDDDDDD-OH (0.5 mg) reduced pulmonary inflammation and the concentration of Mannheimia (Pasteurella) haemolytica in infected lung tissue. Administration of H-DDDDDDD-OH after infection was more effective in reducing the consolidation and lesion scores at the deposition site than its administration prior to infection. Hence, the in vivo effectiveness of AP suggests that it may have applications in the treatment of pulmonary infections. Further studies are needed to confirm these findings and also to determine the optimal doses and intervals of H-DDDDDDD-OH therapy. PMID- 11397621 TI - Continuous versus intermittent administration of ceftazidime in intensive care unit patients with nosocomial pneumonia. AB - A prospective, randomized pilot study was undertaken to compare the efficacy of continuous versus intermittent ceftazidime in ICU patients with nosocomial pneumonia. Ceftazidime was administered either as a 3 g/day continuous infusion (CI) or an intermittent infusion (II) of 2 g every 8 h. In addition, all patients received concomitant once-daily tobramycin. The demographics of the evaluable patients (n = 35) were similar between the groups: age (years), CI 46 +/- 16, II 56 +/- 20; Apache score, CI 14 +/- 4, II 16 +/- 6; time (days) from admission to diagnosis, CI 9 +/- 6, II 9 +/- 6. Clinical efficacy, defined as cure/improvement was similar between groups [n (%), CI 16/17 (94), II 15/18 (83)], while microbiological response was also comparable [n (%), CI 10/13 (76), II 12/15 (80)]. Minimal inhibitory concentrations (MICs) for all isolates were measured throughout the treatment course; there was no development of resistance during therapy for either regimen. While limited clinical data exist, our results suggest that the use of ceftazidime by CI administration maintains clinical efficacy, optimizes the pharmacodynamic profile and uses less antibiotic compared with the standard 2 g every 8 h intermittent dosing regimen. PMID- 11397623 TI - Limits of deep salvage antiretroviral therapy with nelfinavir plus either efavirenz or nevirapine, in highly pre-treated patients with HIV disease. AB - The virological and immunological response to an efavirenz-containing rescue regimen was compared with that of a nevirapine-containing one in a prospective, open-label 12-month study performed in patients previously treated with > or = 12 months of nucleoside analogue monotherapy, and > or = 15 months of indinavir- or ritonavir-containing HAART. Pooled laboratory data were assessed according to change or continuation of nucleoside analogues, at the time of start of salvage treatment. An improvement of markers of HIV disease progression occurred in all 59 evaluable patients (with a higher viral load decrease in the efavirenz over the nevirapine group at the third month), but the virological response was neither complete nor sustained at the end of study, irrespective of efavirenz or nevirapine adjunct, and complete viral suppression was attained in only 16.9% of subjects. A progressively increasing mean CD4+ lymphocyte count characterized the immunological response of all patients. The 35 patients who changed at least one nucleoside analogue when introducing salvage therapy had a better outcome, irrespective of the non-nucleoside reverse transcriptase inhibitor chosen. Rescue strategies are increasingly needed in HIV-infected patients treated for a long time with HAART. The association of nelfinavir, a non-nucleoside reverse transcriptase inhibitor, and dual nucleoside analogues, is popular among patients who fail first-line HAART. When prolonged prior treatment with nucleoside analogue monotherapy and HAART are of concern, and a quite elevated viral load is present, this salvage regimen may not allow a complete and sustained virological response in a 1-year period, while a more favourable immunological recovery can be expected. However, the concurrent change of nucleoside analogues significantly improves treatment outcome. PMID- 11397624 TI - Effects of Helichrysum italicum extract on growth and enzymatic activity of Staphylococcus aureus. AB - Helichrysum italicum G. Don (Compositae) is a shrub commonly found in dry, sandy and stony areas of Mediterranean regions. This plant is known for its anti inflammatory, anti-allergic and antimicrobial activity. The aim of this study was to evaluate the effect of the diethyl ether extract on growth of Staphylococcus aureus (ATCC 6538P, MRSA and MSSA isolates) and the influence of subminimum inhibitory concentrations (subMICs) on some enzymes which are considered virulence factors. The results indicate that the H. italicum extract had an inhibitory effect on S. aureus strains reducing both their growth and some of the enzymes such as coagulase, DNAse, thermonuclease and lipase. Helichrysum italicum extract could be a novel antimicrobial agent, less toxic to human skin and tissues, worthy of further studies. PMID- 11397625 TI - Another look at differences in the susceptibility of Escherichia coli and Klebsiella pneumoniae to cephalothin and cefazolin. AB - The significance of in vitro susceptibility tests on Enterobacteriaceae to cephalothin and cefazolin has not been exactly defined in the guidelines of the National Committee for Clinical Laboratory Standards. In the hope of clarifying this confusion, we provide additional information from an ancillary study of the Taiwan Surveillance of Antimicrobial Resistance 1998 (TSAR I). There were 505 Escherichia coli and 227 Klebsiella pneumoniae isolates susceptible to cephalothin, reported by 42 participating hospitals. The susceptibility of these isolates were re-tested at the Microbial Infections Reference Laboratory using cefazolin, with the result that 72% of the 252 cephalothin-resistant E. coli isolates and 24% of the 41 cephalothin-resistant K. pneumoniae isolates were found to be susceptible to cefazolin. We further surveyed the availability of cephalothin and cefazolin in Pharmacy Departments; all of the TSAR I hospitals had cefazolin available in their pharmacies. The resistance rate of E. coli was significantly lower for 12 hospitals that had cefazolin in both pharmacy and laboratory compared with 11 hospitals that had cefazolin available in pharmacy but cephalothin in laboratory. In addition, for all the hospitals that had cephalothin available for clinical use, the resistance rate was twice as low in two hospitals reporting cefazolin susceptibility as in the seven hospitals reporting cephalothin susceptibility. Our findings suggest that inappropriate selection of cephalothin and cefazolin for susceptibility testing contribute to inaccurate indications of in vivo activity for first generation cephalosporins in the treatment of E. coli infections. PMID- 11397627 TI - Response of gerbera jamesonii acrocarpous callus stimulated by mechanical vibration. AB - Applying different mechanical vibrations, 2-3 Hz frequency, to stimulate the gerbera jamesonii acrocarpous callus, we found that it increases the growth rate and the content of soluble protein but decreases the fluidity of cell membranes. When the frequency was greater than 4 Hz, the effect was contrary to that of 2-3 Hz. In this paper, we also prove the double effects in the plant callus growth and explore the mechanism of the double effects in physical stimulation. PMID- 11397626 TI - In vitro activity of quinolones against isolates from paediatric urinary tract infections from three Venezuelan centres. PMID- 11397628 TI - Preparation and properties of thermosensitive hydrogel microcapsules. AB - Temperature-sensitive hydrophilic gel microcapsules have been newly prepared. That is, poly (L-lysineisopropylamide-terephthalic acid) microcapsules containing water have been obtained by an interfacial polymerization at a water/oil interface between L-lysineisopropylamide and terephthaloyldichloride. The microcapsule changes its size between 33 and 35 degrees C. Under 33 degrees C, the microcapsules are fully spherical and can be redispersed in distilled water, while are aggregated above 35 degrees C. The microcapsules, which are observed to show aggregation above 33 degrees C, can be redispersed by decreasing temperature within a few second. The thermosensitive morphological changes of the microcapsules are thus reversible. Also, it has been shown that the permeability of sodium chloride through the microcapsule membrane changes remarkably between 33 and 35 degrees C, while it is kept almost constant independent of temperature between 25 and 33 degrees C or between 35 and 55 degrees C. The permeability of solutes is higher under 33 degrees C than that above 35 degrees C. Such thermosensitive properties result from the fact that the polymer membrane has isopropylamide groups. That is, L-lysineisopropylamide has a chemical structure similar to N-isopropylacrylamide, the polymer of which, poly (N isopropylacrylamide), is a thermosensitive hydrogel having its phase transition temperature around 33 degrees C. PMID- 11397629 TI - Electrical interaction between two cylinders with an ion-penetrable charged membrane in an oil/water interface. AB - The electrical interaction between two long, parallel cylinders each is covered by an ion-penetrable charged membrane immersed in an oil/water interface is investigated. The effects of contact angle, radius of cylinder, and membrane thickness on the electrical interaction force are examined. The results of numerical simulation reveal that the following conditions lead to a greater electrical interaction force: (i) a larger contact angle, i.e. a larger fraction of a cylinder in the oil phase; (ii) a larger cylinder radius; and (iii) a thinner membrane. For a fixed ionic strength, the electrical interaction force is insensible to the type of electrolytes in the water phase, in general. However, if two cylinders are close enough, then the higher the valence of counterions the greater the electrical interaction force. PMID- 11397630 TI - Novel hydrophobic ligand-containing hydrogel membrane matrix: preparation and application to gamma-globulins adsorption. AB - In this study, phenylalanine as a hydrophobic ligand was covalently attached to the co-monomer methacrylochloride. Then, poly(2-hydroxyethylmethacrylate-co methacrylamidophenyalanine) [poly(HEMA-MAPA)] membranes were prepared by UV initiated photopolymerization of HEMA and methacrylamidophenyalanine. The gamma globulins adsorption onto these affinity membranes from aqueous solutions containing different amounts of gamma-globulins at different pH was investigated in a batch system. The gamma-globulins adsorption capacity of the membranes was increased as the ligand density on the membrane surface increase. The non specific adsorption of the gamma-globulins on the pHEMA membranes was negligible. The adsorption phenomena appeared to follow a typical Langmuir isotherm. The maximum adsorption capacity (q(m)) of the poly(HEMA-MAPA4) membrane for gamma globulins was 2.37 mg g(-1) dry membrane. The equilibrium constant (k(d)) value was found to be 1.61x10(-1) mg ml(-1). More than 87% (up to 100%) of the adsorbed gamma-globulins were desorbed in 120 min in the desorption medium containing 50% ethylene glycol in 1.0 M NaCl. PMID- 11397631 TI - Carrageenan-oligochitosan microcapsules: optimization of the formation process(1). AB - The formation of new microcapsules based on polyelectrolyte complexes between carrageenans and oligochitosan has been investigated. The optimization of the process, which includes the selection of the most suitable solvent and investigation of the influence of reaction conditions on capsule properties, is presented. Iota-carrageenan (1.2-2% wt.) prepared in HEPES buffer was found to be the most suitable for the formation of mechanically stable capsules. These new capsules combine extremely high deformability (>90%) and elasticity with permeability control and can be applied in various bioencapsulation technologies. It has been shown that the reaction time influences the mechanical properties, whereas carrageenan concentration and the temperature during the capsule formation effect both mechanical and porosity characteristic of the membrane. Moreover, the temperature influences the kinetics of the diffusion through the complex iota-carrageenan/oligochitosan membrane. In general egress is faster above the sol-gel transition point, indicating applicability in thermo-induced releasing systems. PMID- 11397632 TI - Determination of the van der Waals, electron donor and electron acceptor surface tension components of static Gram-positive microbial biofilms. AB - A large number of studies have shown the influence of the physico-chemical properties of a surface on microbial adhesion phenomenon. In this study, we considered that the presence of a bacterial biofilm may be regarded as a "conditioning film" that may modify the physico-chemical characteristics of the support, and thus the adhesion capability of planktonic micro-organisms coming into contact with this substratum. In this context, we adapted a protocol for biofilm formation that allows, under our experimental conditions, contact angle measurements, the reference method to determine the energetic surface properties of a substratum. This made it possible to determine the van der Waals, electron acceptor and electron donor properties of static biofilms grown at 25 degrees C on stainless-steel slides with six Gram-positive bacteria isolated in dairy plants. A variance analysis indicated significant effects (P<0.05) of the bacterial strains and of the physiological state of the micro-organisms (planktonic or sessile) on the contact angles. To link the energetic properties of the six biofilms with direct adhesion experiments, we measured the affinity of fluorescent carboxylate-modified polystyrene beads for the different biofilm surfaces. The results correlated best with the electron-acceptor components of the biofilm surface energies, stressing the importance of Lewis acid-base interactions in adhesion mechanisms. PMID- 11397633 TI - Rotational and translational dynamics of lysozyme in water-glycerol solution. AB - In this paper, we report a study of the effect of solvent viscosity on both translational and rotational dynamics of a simple model protein: the egg white lysozyme. For this, we investigated the dynamical properties of lysozyme in mixtures water-glycerol by means of parallel measurements of photon correlation spectroscopy (PCS) and dielectric spectroscopy at radiofrequencies (DS). In the framework of the Debye-Stokes-Einstein theory, the translational and rotational coefficients allow an estimation of hydrodynamic radius of the protein. A decoupling between translational and rotational dynamics, observed as a different estimation of hydrodynamic radius, is reported in the literature for some systems. In order to ascertain if this effect is present also in our sample, we performed PCS and DS measurements on lysozyme-water-glycerol solutions. The content of glycerol was in the range of 0-70% w/w, with a solvent viscosity from 0.9 to about 10 cpoise, and the protein concentration was up to 20 mg ml(-1). The average sizes of lysozyme, obtained by the two methods, are remarkably different at high protein concentrations. However, the values of hydrodynamic radius extrapolated to infinite dilution are coincident and independent of glycerol. These results indicate that the diffusive behavior of lysozyme in the water glycerol mixture is coherent with the Debye-Stokes-Einstein hydrodynamic model. PMID- 11397634 TI - Labelling and binding of poly-(L-lysine) to functionalised gold surfaces. Combined FT-IRRAS and XPS characterisation. AB - We compare herein the interfacial reactivity of self-assembled monolayers (SAMs) of 11-mercaptoundecanoic acid (MUA), 1-undecanethiol (UDT) and 11 mercaptoundecanol (MUD) on gold surfaces towards aqueous solutions of poly-(L lysine) (PL). Liquid-phase labelling of PL with the alkyne dicobalt hexacarbonyl cluster 1 combined with analysis of the substrates by Fourier transform infrared reflection-absorption spectroscopy (FT-IRRAS) and X-ray photoelectron spectroscopy (XPS) revealed that irreversible binding of PL occurred in all cases. However, the mechanism of binding involved differed markedly from one monolayer to the other. The main mode of interaction of PL to MUA SAM was of electrostatic nature between the terminal carboxylate of MUA and the ammonium groups of PL. For a similar number of bound thiolate molecules, the UDT adsorbed layer was found less continuous than the MUA one, allowing a higher fraction of PL to directly bind to the gold surface. As for MUD, very little thiolate molecules were adsorbed, leaving bare gold surface areas for non specific adsorption of PL. PMID- 11397635 TI - Effect of some amino acids and peptides on silicic acid polymerization. AB - The polymerization of silicic acid in aqueous solutions at different pH was followed by the colorimetric molybdosilicate method. The role of four amino acids (serine, lysine, proline and aspartic acid) and the corresponding homopeptides was studied. All four amino acids behave the same way and favor the condensation of silicic acid. Peptides exhibit a stronger catalytic effect than amino acids but they appear to behave in very different ways depending on the nature of side groups and pH. Poly-lysine and poly-proline for instance lead to the precipitation of solid phases containing both silica and peptides. The role of these biomolecules on the polymerization of silicic acid is discussed in terms of electrostatic interactions, hydrogen bonds and solubility. PMID- 11397636 TI - An old epithelial cell never dies, it just agonesces away. AB - The behavior of mammary epithelial cells during aging is dynamic and is likely to have significant implications in the pathogenesis of human breast cancer. The growth of epithelial cells over time was thought to parallel that of their underlying stroma, sequentially undergoing a defined period of growth, followed by senescence and, ultimately, cell crisis or rarely immortalization. Recent findings, however, suggest that the evolution of mammary epithelium at the proliferative and chromosomal levels is distinct from that of stroma, contributing to the neoplastic susceptibilities of epithelial cells. PMID- 11397637 TI - Molecular scaffold protein and cellular responses. AB - Mitogen-activated kinases (MAPK) regulate many diverse cellular processes, including growth, differentiation and responses to stress. The organization of MAPKs through the use of scaffolding proteins is crucial for the selective activation of these kinases by different stimuli. Recent studies identify beta arrestins as members of the family of MAPK scaffold proteins. beta-Arrestins not only shut off signaling by uncoupling G-protein-coupled receptors (GPCRs) from their heterotrimeric G proteins, but also contribute to the specificity of GPCRs signaling by recruiting and activating selective MAPKs. PMID- 11397643 TI - Nuclear receptors, coactivators and chromatin: new approaches, new insights. AB - The actions of lipophilic hormones, including steroids, retinoids, thyroid hormone and vitamin D(3), are mediated through a conserved superfamily of nuclear receptor proteins that function as ligand-regulated, DNA-binding transcriptional activators in the chromatin environment of the nucleus. The ligand-dependent transcriptional activity of nuclear receptors is enhanced by various cofactors that remodel chromatin, acetylate nucleosomal histones and contact the basal transcriptional machinery. The current challenge is to understand the mechanistic details of how interactions among these factors enhance transcription of hormone regulated genes assembled into chromatin. Current biochemical and cell-based methods are providing some important clues. PMID- 11397644 TI - Depression: a major, unrecognized risk factor for osteoporosis? AB - Existing studies of the relationship between depression and osteoporosis have been heterogeneous in their design and use of diagnostic instruments for depression, which might have contributed to the different results on the comorbidity of these two conditions. Nevertheless, these studies reveal a strong association between depression and osteoporosis. Endocrine factors such as depression-induced hypersecretion of corticotropin-releasing hormone and hypercortisolism, hypogonadism, growth hormone deficiency and increased concentration of circulating interleukin 6, might play a crucial role in the bone loss observed in subjects suffering from major depression. PMID- 11397645 TI - Vascular effects of estrogens: arterial protection versus venous thrombotic risk. AB - Mechanisms by which estrogen reduces the risk of arterial disease, while simultaneously increasing the risk of venous thrombosis in postmenopausal women, are not clearly understood. In addition to providing beneficial arterial effects on the lipid profile, estrogen both increases production of nitric oxide and decreases production of endothelin-1 from arterial endothelium, decreases intracellular calcium in arterial smooth muscle and might favor fibrinolysis. All of these effects could act in concert to protect against development of arterial occlusive disease. However, comparable effects on venous endothelium and smooth muscle have not been studied systematically, and although blood elements such as platelets and leukocytes contain estrogen receptors, much remains to be learned about the effect that dose and duration of estrogen-treatment might have upon these cells. An integrative approach to understanding the actions of estrogen on the venous system and the interaction of blood elements with the vascular wall is necessary before new therapeutic interventions will provide arterial protection with no risk of venous thrombosis. PMID- 11397646 TI - Molecular properties of the PTH/PTHrP receptor. AB - The receptor for parathyroid hormone (PTH) and PTH-related protein (PTHrP) is a G protein-coupled receptor (GPCR) that plays a key role in controlling blood Ca(2+) concentration and endochondral bone formation. This review focuses on the molecular mechanisms by which the receptor recognizes the PTH and PTHrP peptide ligands and transmits their signal across the cell membrane. The available data suggest that there are two principal components to the ligand-receptor interaction. First, a docking interaction between the C-terminal portion of PTH(1 34) and the N-terminal extracellular domain of the receptor; and second, a weaker interaction between the N-terminal portion of the ligand and the juxtamembrane region of the receptor, which induces signal transduction. A full understanding of these processes could lead to new PTH/PTHrP receptor ligands that are effective in controlling diseases of bone and mineral metabolism, such as osteoporosis. PMID- 11397647 TI - Signaling by extracellular nucleotides in anterior pituitary cells. AB - Pituitary cells secrete ATP, which acts as an autocrine and/or paracrine extracellular messenger on two families of purinergic receptors: G-protein coupled P2Y receptors (P2YRs) and ion-conducting P2X receptors (P2XRs). Lactotrophs and GH(3)-immortalized cells express the P2Y(2)R subtype. Several P2XR subtypes are expressed in pituitary cells. Gonadotrophs and somatotrophs express P2X(2a)R and P2X(2b)R, which occur as heteromeric channels. Lactotrophs and GH(3) cells express one or more ion-conducting subtypes from among P2X(3)R, P2X(4)R and P2X(7)R in homomeric form. Thyrotrophs and corticotrophs also express P2XRs, but their identification requires further study. Pituitary cells express purinergic P1 receptors, which are activated by adenosine. The A(1)R subtype of these receptors is expressed in melanotrophs and GH(3) cells. In this review, we briefly discuss the expression and coupling of A(1)R and P2Y(2)R, and focus on the expression and Ca(2+) signaling of P2XRs. PMID- 11397648 TI - Diabetes mellitus and diabetes-associated vascular disease. AB - Diabetes-related cardiovascular disease remains the leading cause of death in patients with type 2 diabetes. Hypertension is common among diabetics and has the same pathogenetic mechanisms as insulin resistance, in which the activated renin angiotensin system contributes to the emerging high blood pressure and hyperglycemia. Hyperglycemia is one of the triggering factors for vascular dysfunction and clotting abnormalities and, therefore, for accelerated atherosclerosis in diabetes. Glycated hemoglobin levels, as a reflection of the degree of glycemia, are strongly associated with the risk of cardiovascular disease in diabetics and in the general population. Tight glycemic control, the treatment of dyslipidemia and raised blood pressure, in addition to the use of antiplatelet therapy, all powerfully reduce the risks associated with diabetes. Furthermore, angiotensin-converting enzyme inhibitors might offer additional cardioprotection to diabetics above that provided by blood pressure reduction. PMID- 11397649 TI - Forging the links between metabolism and carcinogenesis. AB - Metabolism plays important roles in chemical carcinogenesis, both good and bad. The process of carcinogen metabolism was first recognized in the first half of the twentieth century and developed extensively in the latter half. The activation of chemicals to reactive electrophiles that become covalently bound to DNA and protein was demonstrated by Miller and Miller [Cancer 47 (1981) 2327]. Today many of the DNA adducts formed by chemical carcinogens are known, and extensive information is available about pathways leading to the electrophilic intermediates. Some concepts about the stability and reactivity of electrophiles derived from carcinogens have changed over the years. Early work in the field demonstrated the ability of chemicals to modulate the metabolism of carcinogens, a phenomenon now described as enzyme induction. The cytochrome P450 enzymes play a prominent role in the metabolism of carcinogens, both in bioactivation and detoxication. The conjugating enzymes can also play both beneficial and detrimental roles. As an example of a case in which several enzymes affect the metabolism and carcinogenicity of a chemical, aflatoxin B1 (AFB1) research has revealed insight into the myriad of reaction chemistry that can occur even with a 1s half-life for a reactive electrophile. Further areas of investigation involve the consequences of enzyme variability in humans and include areas such as genomics, epidemiology, and chemoprevention. PMID- 11397650 TI - Mutations in apoptosis genes: a pathogenetic factor for human disease. AB - Cell death by apoptosis is exerted by the coordinated action of many different gene products. Mutations in some of them, acting at different levels in the apoptosis process, have been identified as cause or contributing factor for human diseases. Defects in the transmembrane tumor necrosis factor receptor 1 (TNF-R1) lead to the development of familial periodic fever syndromes. Mutations in the homologous receptor Fas (also named CD95; Apo-1) are observed in malignant lymphomas, solid tumors and the autoimmune lymphoproliferative syndrome type I (ALPS I). A mutation in the ligand for Fas (Fas ligand; CD95 ligand, Apo-1 ligand), which induces apoptosis upon binding to Fas, was described in a patient with systemic lupus erythematodes and lymphadenopathy. Perforin, an other cytotoxic protein employed by T- and NK-cells for target cell killing, is mutated in chromosome 10 linked cases of familial hemophagocytic lymphohistiocytosis. Caspase 10, a representative of the caspase family of proteases, which plays a central role in the execution of apoptosis, is defect in autoimmune lymphoproliferative syndrome type II (ALPS II). The intracellular pro-apoptotic molecule bcl-10 is frequently mutated in mucosa-associated lymphoid tissue (MALT) lymphomas and various non-hematologic malignancies. The p53, an executioner of DNA damage triggered apoptosis, and Bax, a pro-apoptotic molecule with the ability to perturb mitochondrial membrane integrity, are frequently mutated in malignant neoplasms. Anti-apoptotic proteins like bcl-2, cellular-inhibitor of apoptosis protein 2 (c-IAP2) and neuronal apoptosis inhibitory protein 1 (NAIP1) are often altered in follicular lymphomas, MALT lymphomas and spinal muscular atrophy (SMA), respectively. This article reviews the current knowledge on mutations of apoptosis genes involved in the pathogenesis of human diseases and summarises the gradual transformation of discoveries in apoptosis research into benefits for the clinical management of diseases. PMID- 11397651 TI - Tuberous sclerosis gene products in proliferation control. AB - Two genes, TSC1 and TSC2, have been shown to be responsible for tuberous sclerosis (TSC). The detection of loss of heterozygosity of TSC1 or TSC2 in hamartomas, the growths characteristically occurring in TSC patients, suggested a tumor suppressor function for their gene products hamartin and tuberin. Studies analyzing ectopically modulated expression of TSC2 in human and rodent cells together with the finding that a homolog of TSC2 regulates the Drosophila cell cycle suggest that TSC is a disease of proliferation/cell cycle control. We discuss this question including very recent data obtained from analyzing mice expressing a modulated TSC2 transgene, and from studying the effects of deregulated TSC1 expression. Elucidation of the cellular functions of these proteins will form the basis of a better understanding of how mutations in these genes cause the disease and for the development of new therapeutic strategies. PMID- 11397652 TI - Methods for the spiral Salmonella mutagenicity assay including specialized applications. AB - An automated approach to bacterial mutagenicity testing - the spiral Salmonella assay - was developed to simplify testing and to reduce the labor and materials required to generate dose-responsive mutagenicity information. This document provides the reader with an overview of the spiral assay and a discussion of its application for examining the mutagenic potential of pure compounds, complex environmental mixtures, and interactive effects. Guidelines for performing a routine spiral assay are presented, and alternative test methods intended to overcome a variety of technical difficulties (such as restricted sample availability, sample viscosity or volatility, etc.) are recommended. Methods for the computerized analysis of data and the interpretation of results are discussed. PMID- 11397654 TI - Molecular epidemiology of viral pathogens and tracing of transmission routes: hepatitis-, calici- and hantaviruses. AB - BACKGROUND: The need to rapidly identify new therapeutic drugs and vaccines for clinically important viral infections has resulted in intensive study of the molecular properties of viruses. Modern molecular techniques have provided tools for tracing infections and studying the evolution of viruses. OBJECTIVE STUDY AND DESIGN: Two examples illustrating how modern molecular techniques can be used in clinical virology and molecular epidemiology (hepatitis and caliciviruses), and one example documenting their importance in basic research (hantaviruses) will be discussed. RESULTS AND CONCLUSIONS: Water- and food-borne outbreaks caused by the faeco-orally spread hepatitis A virus (HAV) are common in areas lacking proper sanitation, but they are possible also in countries with low seroprevalence. In water epidemics, the sequence comparisons between the virus from patients and from water have been used successfully. Hepatitis B virus variants are clinically important and challenge the diagnostic tests and prophylactic measures. Some hepatitis C (HCV) genotypes appear to be associated with more severe pathology and others respond better to antiviral treatment. Nosocomial and occupational infections are not rare, and the source can be identified by phylogenetic analysis of nucleotide sequences obtained from the infected individuals. The overwhelming role of Norwalk-like caliciviruses (NLV) in adult diarrhoea and especially in food- and water-borne epidemics has become apparent during the last decade. Methods are under development for detecting these viruses, not only from patient samples and water, but also from other environmental samples (e.g. foodstuff and surface swabs). The analysis of the genetic variation and evolution of the Old World hantaviruses in their carrier rodents has shown that the extent of genetic diversity correlates with geographical distance. As a rule, phylogenetic relationships of hantaviruses resemble those of their rodent hosts, suggesting virus-host co-evolution. Exceptional host-switch events allow a study on still radiating hantavirus species. There is suggestive evidence that natural reassortant hantaviruses are involved in human infection. PMID- 11397655 TI - Diagnosis of HIV infection and laboratory monitoring of its therapy. AB - BACKGROUND: Serological diagnosis of human immunodeficiency virus (HIV) infection became available in 1985, with the rapid increase in sensitivity and specificity of enzyme-linked immunosorbent assays (ELISAs) and the supplement tests. Molecular tests for detection of HIV in the diagnosis of HIV infection in special settings and monitoring of HIV-1 infection followed this. OBJECTIVE AND DESIGN: In this review it is intended to give a brief overview of the diagnosis and monitoring of HIV infection. RESULTS AND CONCLUSION: Serological methods and molecular methods for the detection and quantitation of HIV are discussed. PMID- 11397656 TI - Resistance of human immunodeficiency virus type 1 to reverse transcriptase and protease inhibitors: genotypic and phenotypic testing. AB - Treatment of HIV-1-infected persons with antiretroviral drugs including reverse transcriptase (RT) and protease inhibitors has significantly reduced the rate of HIV and AIDS-related morbidity and mortality. However, these treatments can select for drug-resistant viruses which are associated with poor virologic responses to the antiretroviral therapies and loss of clinical benefit. Drug resistance is conferred by single or several amino acid changes in the pol gene. These mutations can be classified as primary when they directly confer reduced drug susceptibility, or secondary when their influence is primarily on replication capabilities of resistant viruses. Both genotypic and phenotypic methods are used for drug resistance testing. Genotypic assays detect resistance related mutations by sequence analysis or point mutations assays. Phenotypic testing measures drug susceptibility of patient-derived viruses in culture assays. Viruses can be conventionally isolated from peripheral blood lymphocytes, or generated more rapidly through recombination of plasma-derived RT/protease sequences and modified HIV-1 vectors. Phenotypic testing provides direct evidence of resistance, is easy to interpret, but is laborious and expensive. In contrast, genotypic testing provides indirect evidence of resistance, is relatively faster and cheaper, but some complex mutation patterns may be difficult to interpret. Non-culture based phenotypic assays that measure susceptibility of RT activity in plasma to RT inhibitors have been described recently, and provide new tools for rapid phenotypic testing. Resistance testing is currently recommended to help guide the choice of new regimens after treatment failure and for guiding therapy in pregnant women. PMID- 11397657 TI - Diagnosis and prognostic markers of HCMV infection. AB - Human cytomegalovirus (HCMV) infections are frequently observed as well in immunocompetent as in immunocompromised patients. Diagnostic techniques for HCMV detection have greatly improved during the recent years. Detection of HCMV by culture has been bettered by centrifugating samples in a shell vial and by using monoclonal antibody to the immediate early antigen. Detection of antigens in leucocytes was facilitated by using whole blood instead of leucocytes separated by dextran sedimentation. Detection of HCMV recently sharpened by using molecular biology methods mainly based on the detection of the genome. Under certain circumstances, and especially in pregnant women, diagnosis of HCMV infection is essentially based on the detection of IgG and IgM antibodies. However, as IgM antibody is not a marker of primary infection, complementary tests are needed to help in the datation of the infection. Among them, the measurement of IgG avidity greatly improved the diagnosis of primary infections. In immunocompromised patients, very sensitive techniques are needed to diagnose HCMV infections. Detection of antigenemia and HCMV DNA are the methods of choice for an early detection of the infection. Diagnosis of HCMV-organ diseases largely depends on the infected organ and the presence of HCMV in the organ is, sometimes, difficult to interpret. Today, diagnosis of congenital infections is possible by detecting HCMV in the amniotic fluid. However, in order to reliably detect HCMV, amniocentesis must be performed after 21 week's gestation and at least 6 weeks after seroconversion. As all HCMV infections do not induce disease, prognostic markers are needed, as well in immunocompromised patients as in fetuses. Many studies were conducted in immunocompromised patients to find prognostic markers of HCMV infection in order to introduce preemptive therapy when needed. It seems that quantitative determination of HCMV DNA could be very useful to predict HCMV disease. Qualitative determination of mRNA could also be useful as a prognostic marker of HCMV disease. Determination of viral genotypes is more controversial. As for the infected fetuses, little was published about the prediction of sequelae. Some factors, such as viral load in amniotic fluid or in fetal blood or level of IgM antibody in fetal blood, may be predictive of sequelae, but these data require further studies. PMID- 11397658 TI - Progress in understanding cytomegalovirus drug resistance. AB - The molecular basis for cytomegalovirus drug resistance against currently used antivirals comprises two genetic loci. In the case of ganciclovir, mutations in both the UL97 protein kinase and UL64 DNA polymerase can lead to resistance, whereas for cidofovir and foscarnet only mutations in UL54 give rise to resistance. Clinically, resistance strains of cytomegalovirus appear after prolonged periods of antiviral therapy especially when treatment has been interrupted or is at sub-optimal doses. Knowledge of the replication dynamics of cytomegalovirus in vivo can be used to predict the virologic course of patients who develop resistance virus. Using such models, a good agreement between experimentally determined viral load and resistance patterns is observed. PMID- 11397659 TI - Diagnosis of hepatitis B infections and monitoring of treatment. AB - Worldside viral hepatitis is still recognized as a major problem particularly in developing countries. During the past two decades there has been important progress in the field of viral hepatitis; the adaptation of molecular biology techniques to viral hepatitis has proven to be of great utility in the diagnosis of 'classical' hepatitis viruses, in monitoring during treatment, and also in learning more about the 'new' viruses. Here, the progress and pitfalls of serologic and molecular diagnosis techniques for viral hepatitis, unusual profiles and benefits of quantitative DNA/RNA tests will be discussed. PMID- 11397660 TI - Hepatitis B virus infection: resistance to antiviral agents. AB - Chronic hepatitis B virus (HBV) infection imposes an enormous public health burden. In patients with chronic hepatitis and high levels of viral replication, inhibitors of the virus polymerase can reduce serum titre and favourably affect the inflammatory process in the liver. Lamivudine, a reverse transcriptase inhibitor, is the first nucleoside analogue to be licensed for treatment of chronic HBV infection. Treatment effects a rapid and profound decrease of serum virus titre, with attendant clinical benefit. Unfortunately drug-resistant species may emerge after 6 months of suppressive therapy. Lamivudine-resistant species have specific amino acid substitutions in the HBV-encoded polymerase. Emergence of these species is frequently associated with loss of clinical benefit. Published data suggest that lamivudine-resistant species exhibit cross resistance to famciclovir, thereby limiting the potential use of famciclovir with lamivudine as combination therapy. Adefovir is under clinical evaluation for treatment of wild-type and lamivudine-resistant HBV. Preliminary data suggest that adefovir achieves potent inhibition of both species. Studies of drug resistance have followed hot-on-the-heels of the development of potent antiviral therapy for chronic HBV. PMID- 11397661 TI - Detection of hepatitis B virus resistance to antivirals. AB - The development of new nucleoside analogs, that inhibit the HBV reverse transcriptase activity, such as lamivudine, famciclovir and others, has provided recently an alternative to interferon therapy for chronic hepatitis B. However, due to the kinetics of viral replication with a high rate of virus production, a relatively long half-life of virus (CCC) DNA in the nucleus of infected hepatocytes, long-term antiviral therapy with a reverse transcriptase inhibitor is required to eradicate viral infection. Recently, it has been reported that lamivudine therapy for chronic hepatitis B in immune competent patients may be associated with the selection of resistant strains in aproximately 20% of the patients after 12 months of therapy. Sequence analysis of the reverse transcriptase domain of resistant viral strains, at the time of viral breakthrough, revealed the occurrence of mutations located in the YMDD motif within the C domain of the viral enzyme with a methionine to valine (M552V) or to isoleucine (M5521) change. Recent reports on larger series of patients pointed that other mutations residing outside of the C domain but mainly in the B domain of the viral polymerase (L528M) could be associated with these mutations in the YMDD motif. The lamivudine resistant mutants, selected in vivo, can be classified in 2 main groups: group I with a double mutation L528M and M552V, and group II with a single mutation M5521. In vitro studies performed in cell culture showed that these mutants have a decreased replication capacity and are indeed resistant to lamivudine. With the development of new antiviral options, genotyping assays and quantitative determination of viremia with highly sensitive assay are clearly warranted for an optimal monitoring of antiviral therapy of chronic hepatitis B. In view of the experimental and clinical data, the capacity of new antiviral strategies based on combination of new inhibitors, including adefavir and entecavir, with immune modulators needs to be further evaluated in animal models and clinical trials to prevent the emergence of resistant viral strains. PMID- 11397662 TI - Diagnosis of herpes simplex virus infections. PMID- 11397663 TI - Herpes simplex virus antiviral drug resistance--current trends and future prospects. AB - The various manifestations of herpes simplex virus (HSV) have been widely treated using antiviral agents for more than 40 years. Acyclovir (ACV) is the drug that has been most commonly used to date. When tested in cell culture, the majority of isolates of HSV are sensitive to ACV with ED50 values of approximately 0.1 microg/ml. ACV-resistant strains (defined as having ED50>2 microg/ml) are rarely encountered in clinical practice among normal patients (<1% isolates) and there is no firm evidence, to date, that this incidence is increasing. Resistant HSV occurs much more frequently, however, among immunocompromised patients during treatment (approximately 5% isolates) where this is recognised to be an important clinical problem leading to ineffective therapy. In this review it is argued that the rapid establishment of neuronal latency in the normal pathogenesis of HSV is the key to the low incidence of resistance development and leads to some optimism concerning future trends. PMID- 11397664 TI - Diagnosis of hepatitis C virus (HCV) infection and laboratory monitoring of its therapy. AB - BACKGROUND: Just after the identification and characterization of hepatitis C virus (HCV) in 1989, tests for the detection of HCV antibodies or HCV RNA in serum were developed. The enzyme-linked immunosorbent assays (ELISAs) and confirmatory/supplemental analytical antibody tests were improved in sensitivity and specificity with the development of further generations of these assays. Application of molecular tests for detecting, quantifying, and characterization of the infecting virus became very important in management of HCV infection. OBJECTIVE AND DESIGN: This review summarizes the assays developed for the diagnosis and management of HCV infection. Strategies for the diagnosis and monitoring with the advantages and disadvantages of the assays based on the setting and goal are discussed according to data in the literature and our experience. RESULTS: Specific laboratory diagnostic tests for hepatitis C virus infection may be discussed under two titles: (i) Serological antibody tests which detect anti-HCV in serum or plasma; (ii) Molecular tests which detect HCV RNA genome, investigate viral load, and determine the characteristics of the genome. Strategies in different laboratory settings which screen populations with different HCV prevalences vary. CONCLUSIONS: Anti-HCV positive result in a low risk setting such as blood banks should be confirmed with an analytical antibody test. Then a HCV RNA test should be performed on serum of the person with a positive or indeterminate confirmatory test result. On the contrary, anti-HCV positive test result in high-risk population or a situation where HCV infection is suspected, it is likely to be true positive and confirmation with HCV RNA test will be significant. Quantitative HCV RNA test and genotyping should be performed if therapy is considered. PMID- 11397665 TI - The Type 1 growth factor receptors and their ligands considered as a complex system. AB - The Type 1 family of growth factors and their receptors play an important role in normal development, wound healing and in diseases such as cancer. The products of the four receptor genes and the ten genes specifying ligands interact in a complex pattern. Such systems may develop emergent properties which cannot be predicted from a reductionist analysis of the interactions of individual components. New methods of determining these higher level properties are, however, being developed. These include microscopic analysis of live cells expressing fluorescently tagged ligands, receptors and second messengers which can provide positional information on components of the system. Computational simulations of the complex interactions are being developed which can help to predict the properties of the system. PMID- 11397666 TI - EGF mutant receptor vIII as a molecular target in cancer therapy. PMID- 11397667 TI - Cyclooxygenase-2: a target for the prevention and treatment of breast cancer. AB - Cyclooxygenase-2 (COX-2), an inducible prostaglandin synthase, is normally expressed in parts of the kidney and brain. Aberrant COX-2 expression was first reported in colorectal carcinomas and adenomas, and has now been detected in various human cancers, including those of the breast. Strikingly, COX-2 overexpression in murine mammary gland is sufficient to cause tumour formation. To date, the role of COX-2 in tumorigenesis has been most intensively studied in the colon. Thus, the relationship between COX-2 and neoplasia can best be illustrated with reference to intestinal tumorigenesis. Here we consider the potential utility of selective COX-2 inhibitors for the prevention and treatment of breast cancer. Data for cancers of the colon and breast are compared where possible. In addition, the mechanisms by which COX-2 is upregulated in cancers and contributes to tumorigenesis are discussed. Importantly, several recent studies of mammary tumorigenesis in animal models have found selective COX-2 inhibitors to be effective in the prevention and treatment of breast cancer. Clinical trials will be needed to determine whether COX-2 inhibition represents a useful approach to preventing or treating human breast cancer. PMID- 11397668 TI - Phytoestrogens after breast cancer. AB - The current extension of the indications for adjuvant chemotherapy, which predisposes to early menopause, and the media coverage of the benefits of hormone replacement therapy (HRT) have led patients with a history of breast cancer to seek treatments for estrogen deprivation. In breast cancer survivors, most physicians avoid HRT because of concern regarding the potential promotion of growth of occult malignant cells by estrogens, due to the estrogen dependence of breast cancer. Soy phytoestrogens are being promoted as the 'natural alternative' to HRT and have been available without restrictions for several years as nutritional supplements. In this paper, data on the complex mammary effects of phytoestrogens in epidemiological studies, in in vitro studies, as well as in in vivo studies on animal carcinogenesis are reviewed. The potential benefits and risks of phytoestrogens are analyzed, and the prescription of phytoestrogens to postmenopausal women after breast cancer and the coprescription with the anti estrogen tamoxifen are discussed. The absence of controlled trials and technical checking of extraction and titration in these preparations on 'free sale' raise a new problem in terms of public health and justify close reasoning and a cautious attitude of physicians, as well as straight information given to women, especially after breast cancer. PMID- 11397669 TI - Treatment of medullary thyroid carcinoma: an update. AB - Prognosis and treatment effectiveness of medullary thyroid carcinoma (MTC) are largely related to the tumour stage, so that early diagnosis represents an important goal for the management of patients. Recent advances in genetic testing have improved the clinical approach to the familial MTC syndromes. There is general agreement that the primary operation for MTC should obtain the complete removal of the neoplastic tissue in the neck, because any adjuvant treatment has never been proven to be effective. The management of residual/recurrent or metastatic MTC still remains controversial, although a multimodal approach to advanced disease may be of value in palliation or local control of tumour progression. The role of surgery, external radiotherapy, radionuclide therapy and medical treatment, including biological response modifiers and cytotoxic drugs, are reviewed and discussed. PMID- 11397671 TI - [Laboratory support for the diagnosis, misdiagnosis and recognition of thyroid disease]. PMID- 11397672 TI - [Is there sugar in the Alzheimer's disease?]. AB - Epidemiological and immunohistochemical studies focus the interest on the contribution of carbohydrates in the pathophysiology of Alzheimer's disease. Diabetes mellitus increases the risk. In the extracellular (senile) plaques, which contain aggregates of amyloid proteins, and in the neurofibrillary tangles within the cytoplasm of neurons, advanced glycation end products were detected. It is discussed whether it is a cause or an effect of the Alzheimer's disease. The vascular origin of the lesions is also considered. PMID- 11397673 TI - [Burkholderia cepacia: dangers of a phytopathogen organism for patients with cystic fibrosis]. AB - Burkholderia cepacia is an environmental bacterium, capable of colonising vegetal and animal tissues, involved in human opportunist nosocomial infections, and above all, in pulmonary colonisations in patients with cystic fibrosis. In these patients, infection may be followed by a severe deterioration with bacteraemia, leading to death. Moreover, owing to the epidemic spread of some clones within cystic fibrosis communities, strict preventive guidelines have to be instituted. Early detection of Burkholderia cepacia colonisation is therefore essential, and requires the use of selective media. Identification by means of conventional procedures may be problematic, all the more as the previously named Burkholderia cepacia strains have been recently shown to constitute five genomovars (I to V), collectively designated the "cepacia complex", of which only three are classified as new species (II = Burkholderia multivorans; IV = Burkholderia stabilis; V = Burkholderia vietnamiensis). Moreover, closely related species, particularly Burkholderia gladioli, are also involved in cystic fibrosis. Many questions still need clarifications, regarding pathogenic mechanisms and propensity for the cystic fibrosis lung of these organisms. Antimicrobial therapeutic options for B. cepacia complex infections are limited by their innate and acquired antibiotic multiresistance. PMID- 11397674 TI - [Comparative study of adherence of five Candida species to polyvinyl chloride]. AB - Colonization of biomaterial by Candida species could be the first step of Candida infection. We examined the in vitro adherence capabilities of five Candida species to polyvinyl chlorure (PVC). Twenty stains were tested: C. albicans (9), C. glabrata (3), C. tropicalis (3), C. kefyr (4) and C. parapsilosis (1). The adherence to PVC was evaluated by direct microscopic examination. The results indicate that the adherence of C. albicans to PVC is similar to that of C. glabrata and C. tropicalis. Only C. kefyr adheres to a lower degree. We conclude that the adherence capabilities of non-albicans species were similar to that of C. albicans. There is a relationship between the adherence of Candida species and their abilities to colonize the biomaterial. The species, which are more adherent, are those, which are more virulent. PMID- 11397675 TI - [Genetic testing for cystic fibrosis: evaluation of the Elucigene CF20 kit in blood and buccal cells]. AB - Routine determination of mutations in cystic fibrosis requires accurate, rapid, reliable and low-cost methods, permitting the simultaneous detection of multiple mutations. The Elucigene CF20 kit developped by Cellmark Diagnostics, uses multiplex ARMS, which allows the screening for 20 CFTR gene mutations (deltaF508, G542X, N1303K, 1717-1G>A, G551D, W1282X, R553X, deltaI507, 1078delT, 2183AA>G, 3849+10kbC>T, R1162X, 621+1G>T, R334W, R347P, 3659delC, R117H, S1251N, E60X, A455E ) in a work day without specific instrumentation. The kit distinguishes between homozygotes and heterozygotes for deltaF508, but not for rare mutations. The kit detects from 68 to 92% of defective alleles in Caucasians. We evaluate the kit in a blind study in two independent laboratories. Thirty blood samples and thirty mouthwash samples from CF patients, carriers and unaffected individuals were analysed by the Elucigene CF20 kit. All the samples were previously analysed by denaturing gradient gel electrophoresis and sequencing. The Elucigene CF20 kit consists of three multiplexes. Each mutiplex contains ARMS specific primers for six to eight mutations and two control reactions. The absence of the upper control fragment indicates that a repeat test is required. We demonstrated a first time amplification rate of 98.3%: of the 60 samples tested, one required a reamplification. Results compared with the reference method demonstrated that in all cases where one or more of the 20 mutations detected by the kit were present in the test set, the kit accurately identified them. Reproducibility was assessed by repeating the analysis of a blood and mouthwash sample five times. Cross reactivity between R117C and R117H, R117P and R117H, R347P and R347H, deltaI507 and deltaF508, G551D and R553X were evaluated. Only a cross reactivity between R347P and R347H was observed. The kit is specially useful for first line study of patients and carrier identification. PMID- 11397676 TI - [In vitro study of the antimutagenic activity of alphahederin]. AB - Authors have studied the antimutagenic power of alphahederin (a saponin extracted from Hedera helix) versus a clastogenic agent, doxorubicin and an aneugenic agent, carbendazim. We have applied a protocol of incorporation of alphahederin (pretreatment, simultaneous treatment and post treatment) to determine a mechanism of action. According to this protocol, alphahederin induces a significant diminution of the rate of micronuclei wathever the phases of the protocol. These results demonstrate the antimutagenic activity of alphahederin with a mechanism of action, both desmutagenic and bioantimutagenic. PMID- 11397677 TI - [Harmonization of practices: application to the measurement of enzymatic activities used in prevention, screening, diagnosis and therapeutic monitoring]. AB - The large metrological variation (CV, about 25%) observed between laboratories, at the national French level, for the measurement of enzymatic activities results in a loss of efficiency in using laboratory results. Current data show that the standardisation of methods is insufficient to solve this problem and needs to be completed by an harmonisation of the practices including the use of a common reference (calibrator). The present work, carried out by the joint working group between laboratories of the Centres for Periodic Health Examination and the French Society of Clinical Biology (SFBC), deals mainly with the feasibility and evaluation of the improvement of the consistency of the results. Twenty laboratories participated in this study. Five independent surveys were conducted during an height month period. Two enzymes were selected because of their clinical importance and their interest in prevention, screening, diagnosis or epidemiology: ALT (alanine aminotransferase) and GGT (gamma-glutamyltransferase). In each survey three kinds of samples i.e. control sera, candidate calibrators and human serum pools, each of them at two levels of activity (one physiological and the other pathological) were measured in duplicate. The low intra-laboratory imprecision and the high degree of the standardisation of used methods, due to an important effort previously done in this field, permitted to consider a common calibration. The stability and mainly the commutability, i.e. the ability for the candidate calibrator to show a behaviour similar to that of human samples towards the used methods, allowed to reduce the inter-laboratory variation by a half to two third-fold, reaching a coefficient of variation < 5% similar to those observed for cholesterolemia or glycemia. This level of consistency should permit to use common reference limits and common decision limits, after validation of this approach in real practice. The consequences of the harmonisation of practices, extended to the all laboratories, exceed largely the scope of this study. The reduction of the uncertainty and a better approach of the accuracy for the measurement of enzymatic activities should led to a real benefit for the patients in terms of prevention, screening, diagnosis or therapeutic monitoring and consequently for the public health. PMID- 11397679 TI - [The clinical use of biological markers of bone turnover in postmenopausal osteoporosis]. PMID- 11397678 TI - [Biochemical markers of bone remodeling]. PMID- 11397680 TI - [Use of biochemical markers of bone turnover in Paget's disease of bone]. PMID- 11397681 TI - [Biological markers of bone turnover in renal failure]. PMID- 11397682 TI - [Monoclonal and oligoclonal pattern in cerebrospinal fluid and cavernous sinus infiltration: tumoral or infectious process?]. PMID- 11397683 TI - [Kinetics of cardiac troponin I (TnIc) after angioplasty and interest in TnIc in unstable angina]. PMID- 11397684 TI - [Drug-induced agranulocytosis: role of the myelogram and biological tests]. PMID- 11397685 TI - [Analytical evaluation of the amount of specific IgE using Stallergy]. PMID- 11397686 TI - [National Homocysteine Meeting Toulouse, September 21, 2000]. PMID- 11397688 TI - [Assessment of a quality management process in biological laboratories of a teaching hospital]. PMID- 11397687 TI - [Scientific report of the Marcel-Merieux Laboratory]. PMID- 11397689 TI - [Reflections of the National Committee of Medical Biology on the formation of biologists]. PMID- 11397691 TI - Is visceral adiposity the "enemy within"? PMID- 11397692 TI - Phospholipid hydrolytic enzymes in a 'cesspool' of arterial intimal lipoproteins: a mechanism for atherogenic lipid accumulation. PMID- 11397693 TI - 2000 George Lyman Duff Memorial Lecture: atherosclerosis is a liver disease of the heart. AB - The production of apolipoprotein B (apoB)-containing lipoproteins by the liver is regulated by a complex series of processes involving apoB being cotranslationally translocated across the endoplasmic reticulum and assembled into a lipoprotein particle. The translocation of apoB across the endoplasmic reticulum is facilitated by the intraluminal chaperone, microsomal triglyceride transfer protein (MTP). MTP facilitates the translocation and folding of apoB, as well as the addition of lipid to lipid-binding domains (which consist of amphipathic beta sheets and alpha helices). In the absence of MTP or sufficient lipid, apoB exhibits translocation arrest. Thus, apoB translation, translocation, and assembly with lipids to form a core-containing lipoprotein particle occur as concerted processes. Abrogation of >/=1 of these processes diverts apoB into a degradation pathway that is dependent on conjugation with ubiquitin and proteolysis by the proteasome. The nascent core-containing lipoprotein particle that forms within the lumen of the endoplasmic reticulum can be "enlarged" to form a mature very low density lipoprotein particle. Additional studies show that the assembly and secretion of apoB-containing lipoproteins are linked to the cholesterol/bile acid synthetic pathway controlled by cholesterol 7alpha hydroxylase. Studies in cultured cells and transgenic mice indicate that the expression of cholesterol 7alpha-hydroxylase indirectly regulates the expression of lipogenic enzymes through changes in the cellular content of mature sterol response element binding proteins. Oxysterols and bile acids may also act via the ligand-activated nuclear receptors LXR and FXR to link the metabolic pathways controlling energy balance and lipid metabolism to nutritional state. PMID- 11397694 TI - Small GTP-binding protein Ral modulates regulated exocytosis of von Willebrand factor by endothelial cells. AB - Weibel-Palade bodies are endothelial cell-specific organelles, which contain von Willebrand factor (vWF), P-selectin, and several other proteins. Recently, we found that the small GTP-binding protein Ral is present in a subcellular fraction containing Weibel-Palade bodies. In the present study, we investigated whether Ral is involved in the regulated exocytosis of Weibel-Palade bodies. Activation of endothelial cells by thrombin resulted in transient cycling of Ral from its inactive GDP-bound to its active GTP-bound state, which coincided with release of vWF. Ral activation and exocytosis of Weibel-Palade bodies were inhibited by incubation with trifluoperazine, an inhibitor of calmodulin, before thrombin stimulation. Functional involvement of Ral in exocytosis was further investigated by the expression of constitutively active and dominant-negative Ral variants in primary endothelial cells. Introduction of active Ral G23V resulted in the disappearance of Weibel-Palade bodies from endothelial cells. In contrast, the expression of the dominant-negative Ral S28N did not affect the amount of Weibel Palade bodies in transfected cells. These results indicate that Ral is involved in regulated exocytosis of Weibel-Palade bodies by endothelial cells. PMID- 11397695 TI - Receptor for advanced glycation end products mediates inflammation and enhanced expression of tissue factor in vasculature of diabetic apolipoprotein E-null mice. AB - Advanced glycation end products (AGEs) and their cell surface receptor, RAGE, have been implicated in the pathogenesis of diabetic complications. Here, we studied the role of RAGE and expression of its proinflammatory ligands, EN-RAGEs (S100/calgranulins), in inflammatory events mediating cellular activation in diabetic tissue. Apolipoprotein E-null mice were rendered diabetic with streptozotocin at 6 weeks of age. Compared with nondiabetic aortas and kidneys, diabetic aortas and kidneys displayed increased expression of RAGE, EN-RAGEs, and 2 key markers of vascular inflammation, vascular cell adhesion molecule (VCAM)-1 and tissue factor. Administration of soluble RAGE, the extracellular domain of the receptor, or vehicle to diabetic mice for 6 weeks suppressed levels of VCAM-1 and tissue factor in the aorta, in parallel with decreased expression of RAGE and EN-RAGEs. Diabetic kidney demonstrated increased numbers of EN-RAGE-expressing inflammatory cells infiltrating the glomerulus and enhanced mRNA for transforming growth factor-beta, fibronectin, and alpha(1) (IV) collagen. In mice treated with soluble RAGE, the numbers of infiltrating inflammatory cells and mRNA levels for these glomerular cytokines and components of extracellular matrix were decreased. These data suggest that activation of RAGE primes cells targeted for perturbation in diabetic tissues by the induction of proinflammatory mediators. PMID- 11397696 TI - Prorenin accumulation and activation in human endothelial cells: importance of mannose 6-phosphate receptors. AB - ACE inhibitors improve endothelial dysfunction, possibly by blocking endothelial angiotensin production. Prorenin, through its binding and activation by endothelial mannose 6-phosphate (M6P) receptors, may contribute to this production. Here, we investigated this possibility as well as prorenin activation kinetics, the nature of the prorenin-activating enzyme, and M6P receptor independent prorenin binding. Human umbilical vein endothelial cells (HUVECs) were incubated with wild-type prorenin, K/A-2 prorenin (in which Lys42 is mutated to Ala, thereby preventing cleavage by known proteases), M6P-free prorenin, and nonglycosylated prorenin, with or without M6P, protease inhibitors, or angiotensinogen. HUVECs bound only M6P-containing prorenin (K(d) 0.9+/-0.1 nmol/L, maximum number of binding sites [B(max)] 1010+/-50 receptors/cell). At 37 degrees C, because of M6P receptor recycling, the amount of prorenin internalized via M6P receptors was >25 times B(max). Inside the cells, wild-type and K/A-2 prorenin were proteolytically activated to renin. Renin was subsequently degraded. Protease inhibitors interfered with the latter but not with prorenin activation, thereby indicating that the activating enzyme is different from any of the known prorenin-activating enzymes. Incubation with angiotensinogen did not lead to endothelial angiotensin generation, inasmuch as HUVECs were unable to internalize angiotensinogen. Most likely, therefore, in the absence of angiotensinogen synthesis or endocytosis, M6P receptor-mediated prorenin internalization by endothelial cells represents prorenin clearance. PMID- 11397697 TI - Function of GATA transcription factors in induction of endothelial vascular cell adhesion molecule-1 by tumor necrosis factor-alpha. AB - Endothelial vascular cell adhesion molecule-1 (VCAM-1) is expressed in response to cytokine stimulation and plays a critical role in inflammatory reactions. Previously, we developed a novel VCAM-1 inhibitor that acts through a mechanism independent of nuclear factor-kappaB activity. It suppresses the binding activity of GATA proteins in cytokine-stimulated endothelial cells, which may be related to the anti-VCAM-1 induction effect of this drug. In this study, we investigated the role of GATA proteins in the induction of VCAM-1 by tumor necrosis factor alpha (TNF-alpha) in human endothelial cells. The mRNA expression of GATA-6 was increased, whereas GATA-3 mRNA was decreased by TNF-alpha stimulation. Electrophoretic mobility shift assay showed that TNF-alpha stimulation increased the DNA binding of GATA-6 but decreased that of GATA-3. Experiments using protein overexpression or antisense oligonucleotides revealed that GATA-6 potently acts as a positive regulator of VCAM-1 gene transcription. In contrast, overexpression of GATA-3 was able to suppress TNF-alpha-induced VCAM-1 expression. Our results provide evidence of the importance of GATA proteins in the induction of VCAM-1 by TNF-alpha in vascular endothelial cells. The switch from GATA-3 to GATA-6 is taken to be an important transcriptional control event in TNF-alpha induction of VCAM-1. PMID- 11397698 TI - Linked chromosome 16q13 chemokines, macrophage-derived chemokine, fractalkine, and thymus- and activation-regulated chemokine, are expressed in human atherosclerotic lesions. AB - Chemokines are important mediators of macrophage and T-cell recruitment in a number of inflammatory pathologies, and chemokines expressed in atherosclerotic lesions may play an important role in mononuclear cell recruitment and macrophage differentiation. We have analyzed the expression of the linked chromosome 16q13 genes that encode macrophage-derived chemokine (MDC/CCL22), thymus- and activation-regulated chemokine (TARC/CCL17), and the CX(3)C chemokine fractalkine (CX(3)CL1) in primary macrophages and human atherosclerotic lesions by reverse transcription-polymerase chain reaction and immunohistochemistry. We show that macrophage expression of the chemokines MDC, fractalkine, and TARC is upregulated by treatment with the Th2-type cytokines interleukin-4 and interleukin-13. High levels of MDC, TARC, and fractalkine mRNA expression are seen in some, but not all, human arteries with advanced atherosclerotic lesions. Immunohistochemistry shows that MDC, fractalkine, and TARC are expressed by a subset of macrophages within regions of plaques that contain plaque microvessels. We conclude that MDC, fractalkine, and TARC, which are chromosome 16q13 chemokines, could play a role in mononuclear cell recruitment into atherosclerotic lesions and influence the subsequent inflammatory response. Macrophage-expressed chemokines upregulated by interleukin-4 may be useful surrogate markers for the presence of Th2-type immune responses in human atherosclerotic lesions. PMID- 11397699 TI - C-type natriuretic peptide induces redifferentiation of vascular smooth muscle cells with accelerated reendothelialization. AB - We recently reported that C-type natriuretic peptide (CNP) occurs in vascular endothelial cells and acts as a vascular-type natriuretic peptide. In the present study, we stimulated the cGMP cascade in proliferating smooth muscle cells (SMCs), in which particulate guanylate cyclase-B, the specific receptor for CNP, is predominantly expressed, by use of an adenovirus encoding rat CNP cDNA (Ad.CNP). In the Ad.CNP-treated cultured SMCs, CNP caused the growth inhibition of SMCs at G(1) phase with an early increase of p21(CIP1/WAF1) expression and subsequent upregulation of p16(INK4a). The expression of smooth muscle myosin heavy chain-2, which is the molecular marker of highly differentiated SMCs, was reinduced in the Ad.CNP-treated SMCs. The Ad.CNP-treated SMCs also reexpressed particulate guanylate cyclase-A, which shows high affinity to atrial and brain natriuretic peptide and is exclusively expressed in well-differentiated SMCs. CNP, which was overexpressed in rabbit femoral arteries in vivo at the time of balloon injury, significantly suppressed neointimal formation. Furthermore, an enhancement of the expression of smooth muscle myosin heavy chain-2 occurred in the residual neointima. In addition, early regeneration of endothelial cells was observed in the Ad.CNP-infected group. Thus, stimulation of cGMP cascade in proliferating dedifferentiated SMCs can induce growth inhibition and redifferentiation of SMCs with accelerated reendothelialization. PMID- 11397700 TI - HMG-CoA reductase inhibitors prevent migration of human coronary smooth muscle cells through suppression of increase in oxidative stress. AB - In vitro and in vivo evidence of a decrease in vascular smooth muscle cell (SMC) migration induced by 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors has been reported. When added to SMC cultures for 6 hours, the HMG-CoA reductase inhibitors fluvastatin, simvastatin, and pravastatin at 1 micromol/L resulted in a 48%, 50%, and 16% suppression, respectively, of human coronary SMC migration; these reductions mirrored the suppression in oxidative stress induced by 1 micromol/L lysophosphatidylcholine (lyso-PC) of 50%, 53% and 19%, respectively. The hydroxylated metabolites of fluvastatin, M(2) and M(3), at 1 micromol/L also suppressed the enhancement of SMC migration by 58% and 45% and the increase in oxidative stress induced by lyso-PC of 58% and 49%, respectively. Lyso-PC activated phospholipase D and protein kinase C (PKC), and this activation was also suppressed by HMG-CoA reductase inhibitors. The inhibition of phospholipase D and PKC was reversed by 100 micromol/L mevalonate, its isoprenoid derivative, farnesol, and geranylgeraniol but not by 10 micromol/L squalene. Antisense oligodeoxynucleotides at 5 micromol/L to PKC-alpha, but not those to the PKC-beta isoform, suppressed the lyso-PC-mediated increases in SMC migration and oxidative stress. These findings suggest that HMG-CoA reductase inhibitors have direct antimigratory effects on the vascular wall beyond their effects on plasma lipids and that they might exert such antimigratory effects via suppression of the phospholipase D- and PKC (possibly PKC-alpha)-induced increase in oxidative stress, which might in turn prevent significant coronary artery disease. PMID- 11397701 TI - Polylysine as a vehicle for extracellular matrix-targeted local drug delivery, providing high accumulation and long-term retention within the vascular wall. AB - We present the first steps in the elaboration of an approach of extracellular matrix-targeted local drug delivery (ECM-LDD), designed to provide a high concentration, ubiquitous distribution, and long-term retention of a drug within the vessel wall after local intravascular delivery. The approach is based on the concept of a bifunctional drug comprising a "therapeutic effector" and an "affinity vehicle," which should bind to an abundant component of the vessel wall. The aim of the present study was to select molecules suitable for the role of affinity vehicles for ECM-LDD and to study their intravascular delivery and retention ex vivo and in an animal model. By use of fluorescence microscopy, the following molecules were selected on the basis of strong binding to cross sections of human vessels: protamine, polylysine, polyarginine, a glycosaminoglycan-binding peptide from vitronectin, and a synthetic dendrimer. With polylysine as a prototypic affinity vehicle, we showed that after intravascular delivery, polylysine was concentrated throughout a luminal layer of the vascular wall to an extremely high concentration of 20 g/L and was retained therein for at least 72 hours of perfusion without noticeable losses. Low molecular weight (fluorescein) and high molecular weight (hirudin) compounds could be chemically conjugated to polylysine and were retained in the vessel wall after intravascular delivery of the conjugates. In conclusion, by use of the ECM LDD method, an extremely high concentration and long-term retention of locally delivered drug can be reached. Polycationic molecules can be considered as potential affinity vehicles for ECM-LDD. PMID- 11397702 TI - Cultured arterial smooth muscle cells maintain distinct phenotypes when implanted into carotid artery. AB - Cultured arterial smooth muscle cells (SMCs) with distinct phenotypic features have been described by several laboratories; however, it is not presently known whether this phenotypic heterogeneity can be maintained within an in vivo environment. To answer this question, we have seeded into the intima of denuded rat carotid artery 2 SMC populations with well-established distinct biological features, ie, spindle-shaped, not growing in the absence of serum, and well differentiated versus epithelioid, growing in the absence of serum, and relatively undifferentiated, derived from the aortic media of newborn rats (aged 4 days) and old rats (aged >18 months), respectively. We show that these 2 populations maintain their distinct biochemical features (ie, expression of alpha smooth muscle actin, smooth muscle myosin heavy chains, and cellular retinol binding protein-1) in the in vivo environment. The old rat media-derived SMCs continue to produce cellular retinol binding protein-1 but little alpha-smooth muscle actin and smooth muscle myosin heavy chains, whereas the newborn rat media derived SMCs continue to express alpha-smooth muscle actin and smooth muscle myosin heavy chains but no cellular retinol binding protein-1. Our results reinforce the notion of arterial SMC phenotypic heterogeneity and suggest that in our model, heterogeneity is controlled genetically and not by the local environment. PMID- 11397703 TI - Oxidized LDL modulates Bax/Bcl-2 through the lectinlike Ox-LDL receptor-1 in vascular smooth muscle cells. AB - Oxidized low density lipoprotein (Ox-LDL) induces apoptosis in vascular smooth muscle cells (VSMCs), which may increase atherosclerotic plaque instability. In this study, we examined the molecular mechanisms causing the Ox-LDL-induced apoptosis in VSMCs, especially focusing on the involvement of Bax/Bcl-2 and the lectinlike Ox-LDL receptor-1 (LOX-1). In cultured bovine aortic smooth muscle cells (BASMCs), Ox-LDL at high concentrations (>60 microg/mL) induced cell death as demonstrated by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. DNA fragmentation was increased in BASMCs treated with high concentrations of Ox-LDL, indicating that the Ox-LDL-induced cell death in VSMCs was apoptosis. Ox-LDL upregulated LOX-1 expression through phosphorylation of extracellular signal-regulated kinase in BASMCs, and a neutralizing anti-LOX-1 monoclonal antibody, which can block LOX-1-mediated cellular uptake of Ox-LDL, prevented the Ox-LDL-induced apoptosis in BASMCs. This antibody also suppressed the increase in the Bax to Bcl-2 ratio induced by Ox-LDL in BASMCs. Furthermore, LOX-1 expression was well colocalized with Bax expression in the rupture-prone shoulder areas of human atherosclerotic plaques in vivo. LOX-1 may play an important role in Ox-LDL-induced apoptosis in VSMCs by modulating the Bax to Bcl 2 ratio. These molecular mechanisms may be involved in destabilization and rupture of atherosclerotic plaques. PMID- 11397704 TI - Elevated C-reactive protein: another component of the atherothrombotic profile of abdominal obesity. AB - Recent studies have suggested that elevated plasma C-reactive protein (CRP) levels are associated with the features of insulin resistance syndrome. In the present study, we have examined the contribution of body composition measured by hydrostatic weighing and of abdominal adipose tissue (AT) accumulation assessed by computed tomography to the variation in plasma CRP levels associated with atherogenic dyslipidemia of the insulin resistance syndrome in a sample of 159 men, aged 22 to 63 years, covering a wide range of adiposity (body mass index values from 21 to 41 kg/m(2)). Plasma CRP levels showed positive and significant correlations with body fat mass (r=0.41, P<0.0001), waist girth (r=0.37, P<0.0001), and visceral AT accumulation measured by computed tomography at L4 to L5 (r=0.28, P<0.0003). Although CRP levels were associated with plasma insulin levels measured in the fasting state and after a 75-g oral glucose load, no significant correlations were found with plasma lipoprotein levels. Finally, comparison of body fatness, of abdominal fat accumulation, and of the features of the insulin resistance syndrome across quintiles of CRP revealed major differences in body fatness and in indices of abdominal AT accumulation between the lowest and the highest CRP quintiles, whereas no significant differences were found for variables of the plasma lipoprotein-lipid profile. These results suggest that obesity and abdominal AT accumulation are the critical correlates of elevated plasma CRP levels found in men with atherogenic dyslipidemia of the insulin resistance syndrome. PMID- 11397705 TI - Energy restriction and weight loss on very-low-fat diets reduce C-reactive protein concentrations in obese, healthy women. AB - C-reactive protein (CRP) is an inflammatory-response protein that is a strong, independent predictor of cardiovascular mortality. CRP is positively associated with body mass index (BMI). In this study, we investigated the effects of dynamic weight loss on CRP in 83 healthy, obese women (mean BMI, 33.8+/-0.4 kg/m(2); range, 28.2 to 43.8 kg/m(2)). Subjects were placed on very-low-fat, energy restricted diets (5700 kJ, 15% fat) for 12 weeks. Weight, waist and hip circumferences, plasma lipids, glucose, and CRP were measured at baseline and after 12 weeks. CRP was positively associated with BMI (r=0.281, P=0.01) and waist circumference (r=0.278, P=0.01) but was not related to other atherosclerosis risk factors. BMI was significantly different between groups split above or below the median for CRP (34.8+/-0.6 kg/m(2) vs 33.0+/-0.5 kg/m(2), P=0.02). After 12 weeks, weight loss was 7.9+/-0.3 kg. CRP was significantly decreased by 26% (P<0.001), and a correlation was observed between weight loss and the change in CRP (r=0.309, P=0.005). The variance in the change in CRP was partly explained by initial CRP (13.6%), energy intake (5.4%), and percentage weight loss (4.6%, P=0.001). This study confirms recent observations that BMI is associated with CRP, a marker for low-grade systemic inflammation. Furthermore, we observed that CRP was lowered in proportion to weight loss. PMID- 11397706 TI - Genome-wide linkage analysis reveals evidence of multiple regions that influence variation in plasma lipid and apolipoprotein levels associated with risk of coronary heart disease. AB - Results of genome-wide linkage analyses to identify chromosomal regions that influence interindividual variation in plasma lipid and apolipoprotein levels in the Rochester, Minn, population are reported. Analyses were conducted for total cholesterol (total-C), triglycerides (TGs), high density lipoprotein cholesterol (HDL-C), apolipoprotein A-I, apolipoprotein A-II, apolipoprotein B, apolipoprotein C-II, apolipoprotein C-III, apolipoprotein E, the total-C/HDL-C ratio, and the TG/HDL-C ratio. Genotypes were measured for 373 genome-wide marker loci on 1484 individuals distributed among 232 multigeneration pedigrees sampled without regard to health status. LOD scores and estimates of additive genetic variance associated with map locations were obtained by using the variance component method of linkage analysis. No evidence of linkage with genes influencing variation in age served as a negative control. Plasma apolipoprotein E levels and the apolipoprotein E gene served as a positive control (LOD score 4.20). Evidence (LOD score >2.00) was provided that was suggestive of a gene or genes on chromosomes 4 and 5 influencing variation in the apolipoprotein A-II level, on chromosome 12 influencing variation in the apolipoprotein A-I level, and on chromosome 17 influencing variation of total-C/HDL-C. These analyses provide new information about genomic regions in humans that influence interindividual variation in plasma lipid and apolipoprotein levels and serve as a basis for further fine-mapping studies to identify new genes involved in lipid metabolism. PMID- 11397707 TI - Determinants of lipid level variability in French-Canadian children with familial hypercholesterolemia. AB - The wide variability in the biochemical expression of familial hypercholesterolemia (FH) is only partly explained by mutational heterogeneity in the low density lipoprotein receptor (LDLR) gene. In the current study, we measured this biochemical variability in a group of children heterozygous for the >15-kb LDLR gene deletion (n=67) and examined the contribution of apolipoprotein (apo) E and B allelic variations to this phenotypic variability. Variances of total cholesterol (TC), LDL-C, and apoB concentrations and of the ratio of TC to high density lipoprotein cholesterol (HDL-C) were increased in FH subjects compared with controls. However, after taking the means into account, the coefficients of variation showed that the variability of LDL-C and apoB concentrations was smaller for FH than for controls and that the variability of TC and of the ratio TC to HDL-C was similar between both groups. The epsilon2/3 genotype was associated with lower mean TC, LDL-C, and apoB concentrations in FH. The magnitude of this effect was smaller in controls than in FH. Indeed, the percentages of total variance of TC, LDL-C, and apoB attributable to the apoE locus were 19.9%, 18.1%, and 11.8%, respectively, in FH cases and 5.9%, 7.4%, and 6.0%, respectively, in controls. We did not detect any effect of the apoB insertion/deletion polymorphism on lipid traits in FH children. However, in controls, we observed a strong interaction between apoE and apoB genotypes on apoB concentrations and on TC to HDL-C ratios. Our study reemphasizes the important role of apoE in lipid metabolism and illustrates that the effects of allelic variations on lipid traits are context dependent. PMID- 11397708 TI - Point mutation (-69 G-->A) in the promoter region of cholesteryl ester transfer protein gene in Japanese hyperalphalipoproteinemic subjects. AB - Cholesteryl ester transfer protein (CETP) transfers cholesteryl ester (CE) from HDL to apolipoprotein (apo) B-containing lipoproteins and plays a crucial role in reverse cholesterol transport, which is a major protective system against atherosclerosis. Genetic CETP deficiency is the most common cause of a marked hyperalphalipoproteinemia (HALP) in the Japanese, and various mutations have been identified in the coding region as well as in the exon/intron boundaries in the CETP gene. In the present study, we identified a novel mutation in the promoter region of the CETP gene. This mutation was a G-to-A substitution at the -69 nucleotide of the promoter region (-69 G-->A), corresponding to the second nucleotide of the PEA3/ETS binding site (CGGAA) located upstream of the putative TATA box. Four (2.0%) of 196 unrelated subjects with a marked HALP (HDL cholesterol >/=2.59 mmol/L=100 mg/dL) were revealed to be heterozygous for the 69 G-->A mutation, and the allelic frequency of the mutant was 0.0102 in the subjects with a marked HALP. The subjects with the -69 G-->A mutation had low plasma CETP levels. Reporter gene assay showed that this mutation markedly reduced the transcriptional activities in HepG2 cells (8% of wild type). These results suggested that this mutation would be dominant negative. In conclusion, a novel -69 G-->A mutation in the CETP gene causes the decreased transcriptional activity leading to HALP. PMID- 11397709 TI - High-fat, high-cholesterol diet increases the incidence of gastritis in LDL receptor-negative mice. AB - Transgenic and knockout mice are widely used as models for atherogenesis studies. While developing a Helicobacter infection model in LDL receptor-negative (LDLR(-/ )) mice, we noticed that mice fed a high-fat, high-cholesterol diet often contracted gastritis independent of infection. To further investigate this finding, we studied 27 male and 18 female LDLR(-/-) mice fed high-fat, 1% or 1.25% cholesterol diets for 3 to 4 months. The extent of atherosclerosis was morphometrically analyzed in the whole aorta, and the degree of gastric inflammation was scored histologically in hematoxylin-eosin-stained stomach sections. The autoantibody titers to epitopes of oxidized LDL were also measured. Mice fed high-fat, high-cholesterol diets had a significantly higher incidence of gastritis than mice fed normal chow, 62% versus 5%, respectively (P<0.0001). This effect was specific for LDLR(-/-) mice, because no difference in gastritis was found in wild-type mice fed either diet. Animals with gastritis showed slightly more atherosclerosis than animals without gastritis: 16.3+/-6.4% versus 12.8+/ 3.4% in males and 9.4+/-3.5% versus 6.5+/-3.3% in females. Cholesterol-fed mice also had significantly higher IgG autoantibody titers against modified LDL than normal chow-fed animals, but no difference was seen between the gastritis and nongastritis groups. We conclude that the standard high-fat, high-cholesterol diet commonly used in many murine models to induce atherosclerosis increased the incidence of gastritis significantly in LDLR(-/-) mice. PMID- 11397710 TI - Endogenous neutralizing antibodies against platelet-derived growth factor-aa inhibit atherogenesis in the cholesterol-fed rabbit. AB - Previous studies have shown that the B chain of platelet-derived growth factor (PDGF) has an important role in atherogenesis. In this study we have investigated the contribution of PDGF-A chain in cholesterol-induced atherogenesis in the New Zealand White rabbit. High titers of antibodies to PDGF-AA or to platelet cytosolic protein (PCP) were induced in these animals by immunization against recombinant human PDGF-AA or human PCP. Rabbits were then fed a 0.25% to 1% cholesterol-containing diet for 10 weeks to induce atherosclerotic lesions; the rabbits were then humanely killed and perfusion-fixed and their aortas were removed. The extent of atherosclerosis in the thoracic aortas was determined by quantitative morphometry after staining with oil red O. The intimal and medial areas in histological sections taken at the level of the first intercostal branch were quantified by image analysis. Immunization against PDGF-AA and PCP, but not against adjuvant alone, resulted in rising titers of antibodies within 2 weeks, the levels of which reached a plateau by 8 weeks. The antibodies to PDGF-AA were isoform-specific, recognized both human and rabbit PDGF-AA, and neutralized the biological activity of PDGF-AA in vitro. Integrated plasma cholesterol levels were similar in both groups. Compared with nonimmune rabbits (n=10), animals immunized against PDGF-AA (n=10) or PCP (n=10) had significantly smaller areas of the aorta covered by atherosclerotic lesions (24.6+/-5.1% and 18.7+/-4.2%, respectively, vs 34.4+/-4.3%; P<0.05). This was associated with a reduced aortic intimal-medial area ratio in PDGF-AA-immunized (0.009+/-0.006) and PCP-immunized (0.025+/-0.017) rabbits than in nonimmune animals (0.159+/-0.066; P<0.05). These data suggest that PDGF-AA is actively involved in cholesterol-induced atherosclerosis in the rabbit. PMID- 11397711 TI - Enzymatically degraded LDL preferentially binds to CD14(high) CD16(+) monocytes and induces foam cell formation mediated only in part by the class B scavenger receptor CD36. AB - Heterogeneity of peripheral blood monocytes is characterized by specific patterns in the membrane expression of Fc gamma-receptor III (FcgammaRIII/CD16) and the lipopolysaccharide receptor (LPS receptor CD14), allowing discrimination of distinct subpopulations. The aim was to analyze the correlation of these phenotypic differences to the early interaction of freshly isolated monocytes with modified lipoproteins by the use of either enzymatically degraded low density lipoprotein (E-LDL), acetylated low density lipoprotein (ac-LDL), oxidized low density lipoprotein (ox-LDL), or native low density lipoprotein. Highest E-LDL binding was observed on CD14(high) CD16(+) monocytes as determined by flow cytometry, suggesting a selective interaction of E-LDL with distinct subpopulations of monocytes. E-LDL induced rapid foam cell formation both in predifferentiated monocyte-derived macrophages and, in contrast to ac-LDL or ox LDL, also in freshly isolated peripheral blood monocytes. This was accompanied by upregulation of the 2 class B scavenger receptors CLA-1/SR-BI (CD36 and LIMPII Analogous-1/scavenger receptor type B class I) and CD36. Cellular binding and uptake of E-LDL was neither competed by ac-LDL nor the class A scavenger-receptor inhibitor polyinosinic acid but was partially inhibited by an excess of ox-LDL. In predifferentiated monocyte-derived macrophages, an anti-CD36 antibody inhibited cellular binding and uptake of E-LDL by approximately 20%, suggesting that recognition of these hydrolase-modified low density lipoprotein particles is mediated only in part by the class B scavenger receptor CD36. PMID- 11397712 TI - Effect of immune deficiency on lipoproteins and atherosclerosis in male apolipoprotein E-deficient mice. AB - To determine whether T cells and B cells influence lipid metabolism and atherosclerosis, we crossed apolipoprotein E-deficient (apoE degrees ) mice with recombination activating gene 2-deficient (RAG2 degrees ) mice. Total plasma cholesterol levels were approximately 20% higher in male apoE degrees mice compared with the apoE degrees RAG2 degrees mice at 8 weeks of age, and plasma triglyceride levels were 2.5-fold higher in the apoE degrees mice even when plasma cholesterol levels were similar. Male mice with plasma cholesterol levels between 400 and 600 mg/dL at 8 weeks of age were euthanized at 27 and 40 weeks of age. The aortic root lesion area in the apoE degrees RAG2 degrees mice, compared with that in the immune-competent apoE degrees mice, was 81% and 57% smaller at 27 and 40 weeks of age, respectively. In contrast, there was no difference in the size of the brachiocephalic trunk lesions. Similar results were obtained with mice euthanized at 40 weeks of age that had 8-week cholesterol levels between 300 and 399 mg/dL. In apoE degrees RAG2 degrees mice, aortic root atherosclerosis was more profoundly suppressed at lower cholesterol levels. Thus, T and B cells and their products differentially influence the development of atherosclerosis at different sites. We also demonstrate a profound effect of the immune system on plasma lipid homeostasis. PMID- 11397713 TI - Mechanism of endothelial dysfunction in apolipoprotein E-deficient mice. AB - Endothelium-dependent relaxations mediated by NO are impaired in a mouse model of human atherosclerosis. Our objective was to characterize the mechanisms underlying endothelial dysfunction in aortas of apolipoprotein E (apoE)-deficient mice, treated for 26 to 29 weeks with a lipid-rich Western-type diet. Aortic rings from apoE-deficient mice showed impaired endothelium-dependent relaxations to acetylcholine (10(-)(9) to 10(-)(5) mol/L) and Ca(2+) ionophore (10(-)(9) to 10(-)(6) mol/L) and endothelium-independent relaxations to diethylammonium (Z)-1 (N,N-diethylamino)diazen-1-ium-1,2-diolate (DEA-NONOate, 10(-)(10) to 10(-)(5) mol/L) compared with aortic rings from C57BL/6J mice (P<0.05). By use of confocal microscopy of an oxidative fluorescent probe (dihydroethidium), increased superoxide anion (O(2)(-)) production was demonstrated throughout the aortic wall but mainly in smooth muscle cells of apoE-deficient mice. CuZn-superoxide dismutase (SOD) and Mn-SOD protein expressions were unaltered in the aorta exposed to hypercholesterolemia. A cell-permeable SOD mimetic, Mn(III) tetra(4 benzoic acid) porphyrin chloride (10(-)(5) mol/L), reduced O(2)(-) production and partially normalized relaxations to acetylcholine and DEA-NONOate in apoE deficient mice (P<0.05). [(14)C]L-Citrulline assay showed a decrease of Ca(2+) dependent NOS activity in aortas from apoE-deficient mice compared with C57BL/6J mice (P<0.05), whereas NO synthase protein expression was unchanged. In addition, cGMP levels were significantly reduced in the aortas of apoE-deficient mice (P<0.05). Our results demonstrate that in apoE-deficient mice on a Western-type fat diet, impairment of endothelial function is caused by increased production of O(2)(-) and reduced endothelial NO synthase enzyme activity. Thus, chemical inactivation of NO with O(2)(-) and reduced biosynthesis of NO are key mechanisms responsible for endothelial dysfunction in aortas of atherosclerotic apoE deficient mice. PMID- 11397714 TI - Antioxidative and antiatherosclerotic effects of human apolipoprotein A-IV in apolipoprotein E-deficient mice. AB - Mice expressing human apolipoprotein A-IV (apoA-IV) mainly in the intestine were obtained in an apolipoprotein E-deficient (apoE(0)) background (apoA-IV/E(0) mice). Quantification of aortic lesions and plasma lipid determination showed that compared with their control apoE(0) counterparts, the apoA-IV/E(0) mice are protected against atherosclerosis without an increase in HDL cholesterol. Because oxidized lipoproteins play an important role in atherogenesis, we tested whether the protection observed in these animals is accompanied by an in vivo reduction of the oxidation parameters. The lag time in the formation of conjugated dienes during copper-mediated oxidation, the aggregation state of LDL, and the presence of anti-oxidized LDL antibodies were measured. The presence of oxidized proteins in tissues and the presence of oxidation-specific epitopes in heart sections of atherosclerotic lesions were also analyzed. Except for lag time, the results showed that the oxidation parameters were reduced in the apoA-IV/E(0) mice compared with the apoE(0) mice. This suggests that human apoA-IV acts in vivo as an antioxidant. In addition, human apoA-IV accumulation was detected in the atherosclerotic lesions of apoA-IV/E(0) mice, suggesting that apoA-IV may inhibit oxidative damage to local tissues, thus decreasing the progression of atherosclerosis. PMID- 11397715 TI - Pro-oxidant effect of vitamin E in cigarette smokers consuming a high polyunsaturated fat diet. AB - Dietary polyunsaturated fats and vitamin E are associated with reduced risk for atherosclerosis, but in smokers, they could promote lipid oxidation. Therefore, we examined the effects of a high polyunsaturated fat diet and vitamin E supplementation on measures of lipid oxidation in cigarette smokers. Ten subjects who smoked >1 pack of cigarettes per day were sequentially fed the following: a baseline diet in which the major fat source was olive oil, a diet in which the major fat source was high-linoleic safflower oil, and finally, the safflower oil diet plus 800 IU vitamin E per day. LDL oxidation lag time and rate and plasma total F(2)-isoprostanes and prostaglandin F(2alpha) (PGF(2alpha)) were determined after 3 weeks on each diet. The safflower oil diet increased total F(2) isoprostanes from 53.0+/-7.2 to 116.2+/-11.2 nmol/L and PGF(2alpha) from 3.5+/ 0.2 to 5.5+/-0.5 nmol/L, without changing LDL oxidation parameters. Addition of vitamin E prolonged mean LDL oxidation lag time but, paradoxically, further increased F(2)-isoprostanes to 188.2+/-10.9 nmol/L and PGF(2alpha) to 7.8+/-0.4 nmol/L. These data suggest that vitamin E may function as a pro-oxidant in cigarette smokers consuming a high polyunsaturated fat diet. PMID- 11397716 TI - Plasma acylation stimulating protein concentration and subcutaneous adipose tissue C3 mRNA expression in nondiabetic and type 2 diabetic men. AB - We studied the effect of an oral fat load on plasma acylation stimulating protein (ASP) concentrations in 9 lean healthy (age 59+/-2 years, body mass index [BMI] 23.2+/-0.4 kg/m(2); both mean+/-SEM), 9 obese nondiabetic (58+/-2 years, BMI 29.4+/-0.5 kg/m(2)), and 12 type 2 diabetic (60+/-2 years, BMI 29.6+/-1.0 kg/m(2)) men. Because ASP is a cleavage product of complement protein C3 (C3adesArg) and its secretion is regulated by insulin, we also examined the subcutaneous adipose tissue expression of C3 mRNA before and after a 240-minute euglycemic hyperinsulinemic clamp in a subgroup of these men. Plasma ASP concentration and adipose tissue C3 mRNA expression were higher in the obese groups than in the lean men. Plasma ASP concentration did not change significantly after the fat load. Fasting plasma ASP concentration and C3 mRNA expression were correlated negatively with insulin sensitivity and positively with the magnitude of postprandial lipemia in nondiabetic but not in type 2 diabetic men. The expression of C3 mRNA was not regulated by insulin. These data suggest that ASP is associated with whole-body glucose and lipid metabolism in nondiabetic individuals, whereas metabolic disturbances in diabetes may overcome the regulatory role of ASP in lipid and glucose metabolism. PMID- 11397717 TI - Ldl modified by hypochlorous acid is a potent inhibitor of lecithin-cholesterol acyltransferase activity. AB - Modification of low density lipoprotein (LDL) by myeloperoxidase-generated HOCl has been implicated in human atherosclerosis. Incubation of LDL with HOCl generates several reactive intermediates, primarily N-chloramines, which may react with other biomolecules. In this study, we investigated the effects of HOCl modified LDL on the activity of lecithin-cholesterol acyltransferase (LCAT), an enzyme essential for high density lipoprotein maturation and the antiatherogenic reverse cholesterol transport pathway. We exposed human LDL (0.5 mg protein/mL) to physiological concentrations of HOCl (25 to 200 micromol/L) and characterized the resulting LDL modifications to apolipoprotein B and lipids; the modified LDL was subsequently incubated with apolipoprotein B-depleted plasma (density >1.063 g/mL fraction), which contains functional LCAT. Increasing concentrations of HOCl caused various modifications to LDL, primarily, loss of lysine residues and increases in N-chloramines and electrophoretic mobility, whereas lipid hydroperoxides were only minor products. LCAT activity was extremely sensitive to HOCl-modified LDL and was reduced by 23% and 93% by LDL preincubated with 25 and 100 micromol/L HOCl, respectively. Addition of 200 micromol/L ascorbate or N acetyl derivatives of cysteine or methionine completely prevented LCAT inactivation by LDL preincubated with 3.51 g/L) compared with the bottom quintile (<2.57 g/L) was 4.3 (P<0.0001) in the first year of follow-up. RR was reduced to 1.7 in the second year but remained significantly and slightly increased in subsequent years. After adjustment for age and confounding risk factors, the RRs (and 95% confidence intervals) associated with a 1-SD increment of fibrinogen (0.64 g/L) for all-cause, cardiovascular disease, cancer, and other-cause mortality were 1.3 (1.2 to 1.4), 1.2 (1.1 to 1.4), 1.3 (1.2 to 1.5), and 1.3 (1.2 to 1.5), respectively. Preexisting diseases did not influence the significant association of fibrinogen with mortality. There was a significant interaction of fibrinogen with white blood cell count but not with cigarette smoking. We conclude that plasma fibrinogen is an independent risk factor for mortality from a broad spectrum of diseases in elderly men and that this universal effect of fibrinogen on mortality may be mediated partly through inflammation. PMID- 11397722 TI - Interactive effect of PAI-1 4G/5G genotype and salt intake on PAI-1 antigen. AB - Activation of the renin-angiotensin-aldosterone system (RAAS) is associated with increased circulating PAI-1 antigen and increased risk of thrombotic cardiovascular events. A 4G/5G polymorphism located 675 bp upstream from the transcription start site of the PAI-1 gene affects PAI-1 antigen concentrations. To test the hypothesis that PAI-1 4G/5G genotype influences the effect of activation of the RAAS on PAI-1 expression, we measured morning PAI-1 antigen concentrations in 76 subjects with essential hypertension during low (10 mmol/d) and high (200 mmol/d) salt intake. Low salt intake was associated with activation of the RAAS as measured by plasma renin activity (2.3+/-0.2 versus 0.5+/-0.0 ng angiotensin I. mL(-1). h(-1), P<0.001) and aldosterone (529+/-40 versus 145+/-12 pmol/L). PAI-1 antigen concentrations were 17.9+/-2.7, 19.2+/-2.5, and 27.8+/-4.0 ng/mL during high salt intake and 19.2+/-2.7, 21.6+/-2.9, and 38.9+/-7.2 ng/mL during low salt intake in the 5G/5G (n=14), 4G/5G (n=40), and 4G/4G (n=22) groups, respectively. There was a significant effect of both salt intake (F=6.0, P=0.017) and PAI-1 4G/5G genotype (F=7.6, P=0.001) on PAI-1 antigen. More importantly, there was a significant interactive effect (F=7.8, P=0.001) of salt intake and PAI-1 4G/5G genotype on PAI-1 antigen. PAI-1 4G/5G genotype influenced the relationship between serum triglycerides and PAI-1 antigen such that the relationship was significant only in 4G homozygotes during either high (R(2)=0.31, P=0.014) or low (R(2)=0.37, P=0.006) salt intake. This study identifies an important gene-by-environment interaction that may influence cardiovascular morbidity and the response to pharmacological therapies that interrupt the RAAS. PMID- 11397723 TI - Adrenomedullin inhibits angiotensin II-induced expression of tissue factor and plasminogen activator inhibitor-1 in cultured rat aortic endothelial cells. AB - Adrenomedullin (AM) is a potent vasodilating peptide having a variety of pharmacological properties mainly in respect to vascular pathophysiology. We have previously demonstrated that angiotensin II (Ang II) or natriuretic peptides have influence on the expression of tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) in vascular endothelial cells. The aim of this study was to elucidate the effects of AM on TF and PAI-1 mRNA and protein expression in endothelial cells. As a result, AM inhibited Ang II-induced TF and PAI-1 mRNA expression in a dose- and time-dependent manner. Because the expression of TF and PAI-1 mRNA induced by Ang II was attenuated by the increase of intracellular concentrations of cAMP by forskolin and 8-bromo-cAMP and because AM increased the intracellular level of cAMP in rat aortic endothelial cells, it was indicated that the inhibitory effect of AM on the expressions of TF and PAI-1 was mainly mediated by the cAMP-dependent signal transduction. Furthermore, the inhibitory effect of AM on TF and PAI-1 expression was partly attenuated by an NO synthase inhibitor, N(G)-nitro-L-arginine methyl ester. In conclusion, AM is shown to contribute to the regulation of blood coagulation and fibrinolysis by vascular endothelial cells mainly via the cAMP pathway. PMID- 11397724 TI - Effect of acute and chronic tea consumption on platelet aggregation in patients with coronary artery disease. AB - Epidemiological studies suggest that tea consumption is associated with a decreased risk of cardiovascular events, but the mechanisms of benefit remain undefined. Platelet aggregation is a precipitating event in cardiovascular disease, and tea contains antioxidant flavonoids that are known to decrease platelet aggregation in vitro. To test the effect of tea consumption on platelet aggregation, we randomized 49 patients with coronary artery disease to either 450 mL of black tea or water consumed initially, followed by 900 mL of tea or water daily for 4 weeks in a crossover design. Ex vivo platelet aggregation in platelet rich plasma was assessed in response to ADP and thrombin receptor-activating peptide at baseline and 2 hours and 4 weeks after beverage consumption. We observed dose-dependent platelet aggregation in response to each agonist, and neither relation was altered by acute or chronic tea consumption. Plasma flavonoids increased with acute and chronic tea consumption, indicating adequate absorption of tea flavonoids. In conclusion, these results demonstrate that acute and chronic black tea consumption does not affect ex vivo platelet aggregation in patients with coronary artery disease. These findings suggest that an effect of tea flavonoids on platelet aggregation is unlikely to be the explanation for the reduction in risk of cardiovascular events noted in epidemiological studies. PMID- 11397725 TI - Elevated circulating levels of monocyte chemoattractant protein-1 in patients with restenosis after coronary angioplasty. PMID- 11397726 TI - Diabetes black spots and death by postcode. The incidence, and inequity, of diabetes are likely to be worsened by obesity. PMID- 11397727 TI - Euthanasia: where the Netherlands leads will the world follow? No. Legalisation is a diversion from improving care for the dying. PMID- 11397728 TI - Research with children. Paediatric practice needs better evidence--gained in collaboration with parents and children. PMID- 11397729 TI - The new NHS information technology strategy. Technology will change practice. PMID- 11397731 TI - Most GPs would consider resigning from NHS. PMID- 11397730 TI - Drug treatment for obesity. We need more studies in men at higher risk of coronary events. PMID- 11397732 TI - Senior hospital doctors issue warning to new government. PMID- 11397734 TI - UK senior scientists support animal genetic modification. PMID- 11397737 TI - Few incompetent doctors are reported to US national data bank. PMID- 11397738 TI - Tobacco companies exploit women, says WHO. PMID- 11397740 TI - Effect of reducing ambulance response times on deaths from out of hospital cardiac arrest: cohort study. AB - OBJECTIVES: To determine the association between ambulance response time and survival from out of hospital cardiopulmonary arrest and to estimate the effect of reducing response times. DESIGN: Cohort study. SETTING: Scottish Ambulance Service. SUBJECTS: All out of hospital cardiopulmonary arrests due to cardiac disease attended by the Scottish Ambulance Service during May 1991 to March 1998. MAIN OUTCOME MEASURES: Survival rate to hospital discharge and potential improvement from reducing response times. RESULTS: Of 13 822 arrests not witnessed by ambulance crews but attended by them within 15 minutes, complete data were available for 10 554 (76%). Of these patients, 653 (6%) survived to hospital discharge. After other significant covariates were adjusted for, shorter response time was significantly associated with increased probability of receiving defibrillation and survival to discharge among those defibrillated. Reducing the 90th centile for response time to 8 minutes increased the predicted survival to 8%, and reducing it to 5 minutes increased survival to 10-11% (depending on the model used). CONCLUSIONS: Reducing ambulance response times to 5 minutes could almost double the survival rate for cardiac arrests not witnessed by ambulance crews. PMID- 11397742 TI - Excess mortality in a population with diabetes and the impact of material deprivation: longitudinal, population based study. AB - OBJECTIVES: To establish the age and sex specific mortality for people with diabetes in comparison with local and national background populations; to investigate the relationship between mortality and material deprivation in an unselected population with diabetes. DESIGN: Longitudinal study, using a population based district diabetes register. SETTING: South Tees, United Kingdom. PARTICIPANTS: All people known to have diabetes living in Middlesbrough and Redcar and Cleveland local authorities on 1 January 1994. MAIN OUTCOME MEASURE: Death, from any cause, between 1 January 1994 and 31 December 1999. RESULTS: Over the six years of the study 1205 (24.9%) of 4842 participants died. All cause standardised mortality ratios for type 1 diabetes were 641 (95% confidence interval 406 to 962) in women and 294 (200 to 418) in men, and those for type 2 diabetes were 160 (147 to 174) in women and 141 (130 to 152) in men. Cause specific standardised mortality ratios were increased for ischaemic heart disease, cerebrovascular disease, and renal disease; no reductions in mortality from other causes were seen. The risk of premature death increased significantly with increasing material deprivation (P<0.001). CONCLUSIONS: Diabetes is associated with excess mortality, even in an area with high background death rates from cardiovascular disease. This excess mortality is evident in all age groups, most pronounced in young people with type 1 diabetes, and exacerbated by material deprivation. Aggressive approaches to the management of cardiovascular risk factors could reduce the excess mortality in people with diabetes. PMID- 11397743 TI - Do doctors position defibrillation paddles correctly? Observational study. PMID- 11397744 TI - Differences in therapeutic consequences of exercise testing between a rural and an urban Danish county: population based study. PMID- 11397745 TI - Cost effectiveness of computer tailored and non-tailored smoking cessation letters in general practice: randomised controlled trial. AB - OBJECTIVES: To develop and evaluate, in a primary care setting, a computerised system for generating tailored letters about smoking cessation. DESIGN: Randomised controlled trial. SETTING: Six general practices in Aberdeen, Scotland. PARTICIPANTS: 2553 smokers aged 17 to 65. INTERVENTIONS: All participants received a questionnaire asking about their smoking. Participants subsequently received either a computer tailored or a non-tailored, standard letter on smoking cessation, or no letter. MAIN OUTCOME MEASURES: Prevalence of validated abstinence at six months; change in intention to stop smoking in the next six months. RESULTS: The validated cessation rate at six months was 3.5% (30/857) (95% confidence interval 2.3% to 4.7%) for the tailored letter group, 4.4% (37/846) (3.0% to 5.8%) for the non-tailored letter group, and 2.6% (22/850) (1.5% to 3.7%) for the control (no letter) group. After adjustment for significant covariates, the cessation rate was 66% greater (-4% to 186%; P=0.07) in the non-tailored letter group than that in the no letter group. Among participants who smoked <20 cigarettes per day, the cessation rate in the non tailored letter group was 87% greater (0% to 246%; P=0.05) than that in the no letter group. Among heavy smokers who did not quit, a 76% higher rate of positive shift in "stage of change" (intention to quit within a particular period of time) was seen compared with those who received no letter (11% to 180%; P=0.02). The increase in cost for each additional quitter in the non-tailored letter group compared with the no letter group was pound 89. CONCLUSIONS: In a large general practice, a brief non-tailored letter effectively increased cessation rates among smokers. A tailored letter was not effective in increasing cessation rates but promoted shift in movement towards cessation ("stage of change") in heavy smokers. As a pragmatic tool to encourage cessation of smoking, a mass mailing of non-tailored letters from general practices is more cost effective than computer tailored letters or no letters. PMID- 11397747 TI - A survey of validity and utility of electronic patient records in a general practice. AB - OBJECTIVE: To develop methods of measuring the validity and utility of electronic patient records in general practice. DESIGN: A survey of the main functional areas of a practice and use of independent criteria to measure the validity of the practice database. SETTING: A fully computerised general practice in Skipton, north Yorkshire. SUBJECTS: The records of all registered practice patients. MAIN OUTCOME MEASURES: Validity of the main functional areas of the practice clinical system. Measures of the completeness, accuracy, validity, and utility of the morbidity data for 15 clinical diagnoses using recognised diagnostic standards to confirm diagnoses and identify further cases. Development of a method and statistical toolkit to validate clinical databases in general practice. RESULTS: The practice electronic patient records were valid, complete, and accurate for prescribed items (99.7%), consultations (98.1%), laboratory tests (100%), hospital episodes (100%), and childhood immunisations (97%). The morbidity data for 15 clinical diagnoses were complete (mean sensitivity=87%) and accurate (mean positive predictive value=96%). The presence of the Read codes for the 15 diagnoses was strongly indicative of the true presence of those conditions (mean likelihood ratio=3917). New interpretations of descriptive statistics are described that can be used to estimate both the number of true cases that are unrecorded and quantify the benefits of validating a clinical database for coded entries. CONCLUSION: This study has developed a method and toolkit for measuring the validity and utility of general practice electronic patient records. PMID- 11397749 TI - Extracts from "Clinical Evidence". Obesity. PMID- 11397751 TI - ABC of AIDS. Antiretroviral drugs. PMID- 11397753 TI - Implementing clinical governance: turning vision into reality. PMID- 11397754 TI - Who should measure quality of life? PMID- 11397756 TI - Cycle injury trends: helmets are most likely explanation. PMID- 11397757 TI - Computer assisted learning aids management of course information. PMID- 11397758 TI - Patients with depression can be taught how to improve recovery. PMID- 11397759 TI - Prevalence of type 2 diabetes in children in Birmingham. PMID- 11397760 TI - Conclusions about type 1 diabetes and hygiene hypothesis are premature. PMID- 11397761 TI - What author really said about malaria and climate change. PMID- 11397766 TI - A tale of seven surgeons. PMID- 11397771 TI - New era for translational research in cardiac arrhythmias. PMID- 11397772 TI - Three things you should know when considering the atria: location, location, location. PMID- 11397773 TI - Harmonic interplay of angiogenic growth factors in the development of coronary blood vessels. PMID- 11397774 TI - Human urotensin II-induced contraction and arterial smooth muscle cell proliferation are mediated by RhoA and Rho-kinase. AB - The aim of this work was to investigate the coupling of human urotensin II (hU II) to RhoA activation and regulation of RhoA-dependent functions. The use of the Rho-kinase inhibitor Y-27632 and the development of a membrane-permeant RhoA inhibitor (TAT-C3) allowed us to demonstrate that hU-II induced arterial smooth muscle contraction, actin stress fiber formation, and proliferation through the activation of the small GTPase RhoA and its downstream effector Rho-kinase. PMID- 11397775 TI - Circulation Research: origin and early years. AB - Circulation Research, first published in 1953, was created by the American Heart Association as "the authoritative new journal for investigators of the basic sciences as they apply to the heart and circulation." This review of the early years of the journal highlights the contributions of the first four Editors: Carl J. Wiggers, Carl F. Schmidt, Eugene M. Landis, and Julius H. Comroe, Jr. The success of Circulation Research is seen not only in the high quality of the articles published in its pages but also in the remarkable improvements in prevention and treatment of cardiovascular disease that have occurred over the past half century. PMID- 11397776 TI - Integrins and the myocardium. AB - Extracellular matrix provides a structural, chemical, and mechanical substrate that is essential in cardiac development, growth, and responses to pathophysiological signals. Transmembrane receptors termed integrins provide a dynamic interaction of environmental cues and intracellular events. Integrins orchestrate multiple functions in the intact organism including organogenesis, regulation of gene expression, cell proliferation, differentiation, migration, and death. They are expressed in all cellular components of the cardiovascular system, including the vasculature, blood, cardiac myocytes and nonmuscle cardiac cells. The focus of this review will be on the role of integrins in the myocardium. We will provide background on integrin structure and function, discuss how the expression of integrins is critical to the form and function of the developing and postnatal myocardium, and review the known data on integrins as signaling molecules in the heart. Finally, we will offer insights to the future research directions into this important family of extracellular matrix receptors in the myocardium. PMID- 11397777 TI - Gene transfer of dominant-negative mutants of extracellular signal-regulated kinase and c-Jun NH2-terminal kinase prevents neointimal formation in balloon injured rat artery. AB - We previously reported that extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK), belonging to mitogen-activated protein kinases, are rapidly activated in balloon-injured artery. Therefore, we examined the role of these kinase activations in neointimal formation by using an in vivo gene transfer technique. We made the dominant-negative mutants of ERK (DN-ERK) and JNK (DN-JNK) to specifically inhibit endogenous ERK and JNK activation, respectively. Before balloon injury, these mutants were transfected into rat carotid artery using the hemagglutinating virus of Japan liposome method. In vivo transfection of DN-ERK and DN-JNK significantly suppressed the activation of ERK and JNK, respectively, after balloon injury, confirming successful expression of the transfected genes. Neointimal formation at 14 and 28 days after injury was prevented by gene transfer of DN-ERK or DN-JNK. Furthermore, bromodeoxyuridine labeling index and total cell-counting analysis at 7 days showed that either DN ERK or DN-JNK remarkably suppressed smooth muscle cell (SMC) proliferation in both the intima and the media after injury. Gene transfer of wild-type ERK (W ERK) or JNK (W-JNK) significantly enhanced neointimal hyperplasia at 14 days after injury. Furthermore, DN-ERK and DN-JNK significantly suppressed serum induced SMC proliferation in vitro. We obtained the first evidence that in vivo gene transfer of DN-ERK or DN-JNK prevented neointimal formation in balloon injured artery by inhibiting SMC proliferation. Thus, ERK and JNK activation triggers SMC proliferation, leading to neointimal formation. These kinases may be the new therapeutic targets for prevention of vascular diseases. PMID- 11397778 TI - Recruitment of serum response factor and hyperacetylation of histones at smooth muscle-specific regulatory regions during differentiation of a novel P19-derived in vitro smooth muscle differentiation system. AB - Little is known regarding transcriptional regulatory mechanisms that control the sequential and coordinate expression of genes during smooth muscle cell (SMC) differentiation. To facilitate mechanistic studies of SMC differentiation, we established a novel P19-derived clonal cell line (designated A404) harboring a smooth muscle (SM) alpha-actin promoter/intron-driven puromycin resistance gene. Retinoic acid plus puromycin treatment stimulated rapid differentiation of multipotential A404 cells into SMCs that expressed multiple SMC differentiation marker genes, including the definitive SM-lineage marker SM myosin heavy chain. Using this system, we demonstrated that various transcription factors were upregulated coincidentally with the expression of SMC differentiation marker genes. Of interest, the expression of serum response factor (SRF), whose function is critical for SMC-specific transcription, was high in undifferentiated A404 cells, and it did not increase over the course of differentiation. However, chromatin immunoprecipitation analyses showed that SRF did not bind the target sites of endogenous SMC marker genes in chromatin in undifferentiated cells, but it did in differentiated A404 cells, and it was associated with hyperacetylation of histones H3 and H4. The present studies define a novel cell system for studies of transcriptional regulation during the early stages of SMC differentiation, and using this system, we obtained evidence for the involvement of chromatin remodeling and selective recruitment of SRF to CArG elements in the induction of cell-selective marker genes during SMC differentiation. PMID- 11397779 TI - Vascular endothelial growth factor and basic fibroblast growth factor differentially modulate early postnatal coronary angiogenesis. AB - The roles of vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF [FGF-2]) in early postnatal regulation of coronary angiogenesis were investigated by administering neutralizing antibodies to these growth factors between postnatal days 5 and 12. Immunohistochemistry and Western blotting both revealed decreases in VEGF protein in the hearts of rats treated with either antibody. In contrast, bFGF mRNA increased in both treated groups, whereas VEGF mRNA was unchanged. Using stereological assessment of perfusion fixed hearts, we found that both anti-VEGF and anti-bFGF inhibited the rapid and marked capillary growth that occurs during this time period and that the effects of the two neutralizing antibodies are not additive. Arteriolar growth, as indicated by a lower length density, was inhibited by anti-bFGF, but not anti VEGF. When both anti-VEGF and anti-bFGF were administered, arteriolar length density was not significantly lower, but the population of small arterioles (<15 microm) was markedly reduced, whereas the percentage of large arterioles (26 to 50 microm) more than doubled. Thus, inhibition of both growth factors negated or limited the formation of small arterioles and facilitated an expansion of the largest arterioles. These in vivo data are the first to document that during the early neonatal period, (1) both VEGF and bFGF modulate capillary growth, (2) bFGF facilitates arteriolar growth, and (3) the two growth factors interact to establish the normal hierarchy of the arteriolar tree. PMID- 11397780 TI - Clock genes in the heart: characterization and attenuation with hypertrophy. AB - We investigated whether the heart, like other mammalian organs, possesses internal clocks, and, if so, whether pressure overload-induced hypertrophy alters the clock mechanism. Clock genes are intrinsically maintained, as shown by rhythmic changes even in single cells. Clocks are believed to confer a selective advantage by priming the cell for the expected environmental stimulus. In this way, clocks allow anticipation, thereby synchronizing responsiveness of the cell with the timing of the stimulus. We have found that in rat heart all mammalian homologues of known Drosophila clock genes (bmal1, clock, cry1, cry2, per1, per2, per3, dbp, hlf, and tef) show circadian patterns of expression and that the induction of clock output genes (the PAR [rich in proline and acidic amino acid residues] transcription factors dbp, hlf, and tef) is attenuated in the pressure overloaded hypertrophied heart. The results expose a new dynamic regulatory system in the heart, which is partially lost with hypertrophy. Although the target genes of these PAR transcription factors are not known in the heart, the results provide evidence for a diminished ability of the hypertrophied heart to anticipate and subsequently adapt to physiological alterations during the day. PMID- 11397781 TI - Coupled gating between cardiac calcium release channels (ryanodine receptors). AB - Excitation-contraction coupling in heart muscle requires the activation of Ca(2+) release channels/type 2 ryanodine receptors (RyR2s) by Ca(2+) influx. RyR2s are arranged on the sarcoplasmic reticular membrane in closely packed arrays such that their large cytoplasmic domains contact one another. We now show that multiple RyR2s can be isolated under conditions such that they remain physically coupled to one another. When these coupled channels are examined in planar lipid bilayers, multiple channels exhibit simultaneous gating, termed "coupled gating." Removal of the regulatory subunit, the FK506 binding protein (FKBP12.6), functionally but not physically uncouples multiple RyR2 channels. Coupled gating between RyR2 channels may be an important regulatory mechanism in excitation contraction coupling as well as in other signaling pathways involving intracellular Ca(2+) release. PMID- 11397782 TI - Arrhythmogenesis and contractile dysfunction in heart failure: Roles of sodium calcium exchange, inward rectifier potassium current, and residual beta adrenergic responsiveness. AB - Ventricular arrhythmias and contractile dysfunction are the main causes of death in human heart failure (HF). In a rabbit HF model reproducing these same aspects of human HF, we demonstrate that a 2-fold functional upregulation of Na(+)-Ca(2+) exchange (NaCaX) unloads sarcoplasmic reticulum (SR) Ca(2+) stores, reducing Ca(2+) transients and contractile function. Whereas beta-adrenergic receptors (beta-ARs) are progressively downregulated in HF, residual beta-AR responsiveness at this critical HF stage allows SR Ca(2+) load to increase, causing spontaneous SR Ca(2+) release and transient inward current carried by NaCaX. A given Ca(2+) release produces greater arrhythmogenic inward current in HF (as a result of NaCaX upregulation), and approximately 50% less Ca(2+) release is required to trigger an action potential in HF. The inward rectifier potassium current (I(K1)) is reduced by 49% in HF, and this allows greater depolarization for a given NaCaX current. Partially blocking I(K1) in control cells with barium mimics the greater depolarization for a given current injection seen in HF. Thus, we present data to support a novel paradigm in which changes in NaCaX and I(K1), and residual beta AR responsiveness, conspire to greatly increase the propensity for triggered arrhythmias in HF. In addition, NaCaX upregulation appears to be a critical link between contractile dysfunction and arrhythmogenesis. PMID- 11397783 TI - Potential ionic mechanism for repolarization differences between canine right and left atrium. AB - Experimental and clinical evidence suggests a critical role for the left atrium (LA) in atrial fibrillation (AF). In animal models, repolarization is faster in the LA than in the right atrium (RA), leading to more stable reentry circuits with a shorter intrinsic period in the LA. The ionic mechanisms underlying LA-RA repolarization differences are unknown. Therefore, we evaluated ionic currents and action potentials (APs) with the whole-cell patch clamp in isolated canine atrial myocytes. The density of the rapid delayed rectifier current (I(Kr)) was greater in the LA (eg, 1.83+/-0.10 pA/pF at +20 mV) than in the RA (1.15+/-0.07 pA/pF, P<0.01; n=16 cells per group). The slow and ultrarapid delayed rectifier, the inward rectifier, L-type Ca(2+), and transient outward K(+) currents were all comparable in the LA and RA. There were no differences in kinetic or voltage dependent properties of currents in LA versus RA. Western blots of ether-a-go-go related gene (ERG) protein in three RA and corresponding LA regions showed significantly greater ERG expression in LA. AP duration (APD) was shorter in the LA versus RA in both isolated cells and multicellular preparations, and the effective refractory period (ERP) was shorter in the LA compared with the RA in vivo. Dofetilide had significantly larger APD- and ERP-increasing effects in the LA compared with RA, and LA-RA repolarization differences were eliminated by exposure to dofetilide. We conclude that LA myocytes have larger I(Kr) than do RA myocytes, contributing importantly to the shorter APD and ERP in LA. The larger LA I(Kr) may participate in the ability of the LA to act as a "driver region" for AF, with potentially important implications for understanding AF mechanisms and antiarrhythmic therapy. PMID- 11397784 TI - Role of intracellular Na(+) kinetics in preconditioned rat heart. AB - To elucidate the role of intracellular Na(+) kinetics in the mechanism for ischemic preconditioning (IPC), we measured intracellular Na(+) concentration ([Na(+)](i)) using (23)Na-magnetic resonance spectroscopy in isolated rat hearts. IPC significantly delayed the initial [Na(+)](i) increase (d[Na(+)](i)/dt) compared with non-IPC control, resulting in attenuation of Na(+) accumulation (Delta[Na(+)](i)) during 27 minutes of ischemia with better functional recovery. [Na(+)](i) in IPC, but not in control, recovered to preischemic level during a 6 minute reperfusion. The Na(+)-H(+) exchange inhibitor further suppressed d[Na(+)](i)/dt in both control and IPC hearts with concomitant improvement of functional recovery, suggesting little contribution to the mechanism of IPC. The mitochondrial ATP-sensitive K(+) (mito K(ATP)) channel activator diazoxide (30 micromol/L) completely mimicked both [Na(+)](i) kinetics and functional recovery in IPC without any additive effects to IPC. The mito K(ATP) channel blocker 5 hydroxydecanoic acid (100 micromol/L) lost protective effect as well as the attenuation of d[Na(+)](i)/dt and [Na(+)](i) recovery induced by diazoxide. However, 5-hydroxydecanoic acid also lost IPC-induced protection, but incompletely abolished the alteration of d[Na(+)](i)/dt and the [Na(+)](i) recovery. The Na(+)/K(+)-ATPase inhibitor ouabain (200 micromol/L) did not change d[Na(+)](i)/dt in non-IPC hearts, but it abolished the IPC- or diazoxide-induced reduction of d[Na(+)](i)/dt and the [Na(+)](i) recovery, whereas IPC followed by ouabain treatment showed partial functional recovery with smaller Delta[Na(+)](i) than other ouabain groups. In conclusion, alteration of Na(+) kinetics by preserving Na(+) efflux via Na(+)/K(+)-ATPase mediated by mito K(ATP) channel activation mainly contributes to functional protection in IPC hearts. The contribution of mito K(ATP) channel-independent pathway relating to Na(+) kinetics including reduced Na(+) influx is limited in functional protection of IPC. PMID- 11397785 TI - Sodium-coupled glucose transporter as a functional glucose sensor of retinal microvascular circulation. AB - To clarify the function of the Na(+)-coupled glucose transporter in the regulation of cellular tone of cultured retinal pericytes, we investigated the effects of extracellular glucose concentration on cell size. The surface area and diameter of cultured bovine retinal pericytes under different glucose concentrations were measured by using a light microscope with a digital camera. We also examined the effects of extracellular Na(+) and Ca(2+), inhibitors of the Na(+)-coupled glucose transporter and Na(+)-Ca(2+) exchanger, a Ca(2+) channel blocker, and nonmetabolizable sugars on cell size. The surface area and diameter of the cells changed according to extracellular glucose concentrations. alpha Methyl glucoside, which enters the cell through the Na(+)-coupled glucose transporter, induced cellular contraction. However, the cells did not contract in response to 2-deoxyglucose, which enters the cell through a facilitated glucose transporter. Glucose-induced cellular contraction was abolished in the absence of extracellular Na(+) and Ca(2+). Moreover, phlorizin, an inhibitor of the Na(+) coupled glucose transporter, and 2',4'-dichlorobenzamil-HCl, an inhibitor of the Na(+)-Ca(2+) exchanger, also abolished glucose-induced cellular contraction, whereas nicardipine, a Ca(2+) channel blocker, did not. Our results indicate that high extracellular glucose concentrations induce contraction of bovine retinal pericytes via Na(+) entry through a Na(+)-coupled glucose transporter, suggesting that the Na(+)-coupled glucose transporter may act as a functional glucose sensor of retinal microvascular circulation.> PMID- 11397786 TI - Mechanisms underlying the reentrant circuit of atrioventricular nodal reentrant tachycardia in isolated canine atrioventricular nodal preparation using optical mapping. AB - The reentrant pathways underlying different types of atrioventricular (AV) nodal reentrant tachycardia have not yet been elucidated. This study was performed to optically map Koch's triangle and surrounding atrial tissue in an isolated canine AV nodal preparation. Multiple preferential AV nodal input pathways were observed in all preparations (n=22) with continuous (73%, n=16) and discontinuous (27%, n=6) AV nodal function curves (AVNFCs). AV nodal echo beats (EBs) were induced in 54% (12/22) of preparations. The reentrant circuit of the slow/fast EB (36%, n=8) started as a block in fast pathway (FP) and a delay in slow pathway (SP) conduction to the compact AV node, then exited from the AV node to the FP, and rapidly returned to the SP through the atrial tissue located at the base of Koch's triangle. The reentrant circuit of the fast/slow EB (9%, n=2) was in an opposite direction. In the slow/slow EB (9%, n=2), anterograde conduction was over the intermediate pathway (IP) and retrograde conduction was over the SP. Unidirectional conduction block occurred at the junction between the AV node and its input pathways. Conduction over the IP smoothed the transition from the FP to the SP, resulting in a continuous AVNFC. A "jump" in AH interval resulted from shifting of anterograde conduction from the FP to the SP (n=4) or abrupt conduction delay within the AV node through the FP (n=2). These findings indicate that (1) multiple AV nodal anterograde pathways exist in all normal hearts; (2) atrial tissue is involved in reentrant circuits; (3) unidirectional block occurs at the interface between the AV node and its input pathways; and (4) the IP can mask the existence of FP and SP, producing continuous AVNFCs. PMID- 11397787 TI - Null mutation of connexin43 causes slow propagation of ventricular activation in the late stages of mouse embryonic development. AB - Connexin43 (Cx43) is the principal connexin isoform in the mouse ventricle, where it is thought to provide electrical coupling between cells. Knocking out this gene results in anatomic malformations that nevertheless allow for survival through early neonatal life. We examined electrical wave propagation in the left (LV) and right (RV) ventricles of isolated Cx43 null mutated (Cx43(-/-)), heterozygous (Cx43(+/)(-)), and wild-type (WT) embryos using high-resolution mapping of voltage-sensitive dye fluorescence. Consistent with the compensating presence of the other connexins, no reduction in propagation velocity was seen in Cx43(-/-) ventricles at postcoital day (dpc) 12.5 compared with WT or Cx43(+/)(-) ventricles. A gross reduction in conduction velocity was seen in the RV at 15.5 dpc (in cm/second, mean [1 SE confidence interval], WT 9.9 [8.7 to 11.2], Cx43(+/)(-) 9.9 [9.0 to 10.9], and Cx43(-/-) 2.2 [1.8 to 2.7; P<0.005]) and in both ventricles at 17.5 dpc (in RV, WT 8.4 [7.6 to 9.3], Cx43(+/)(-) 8.7 [8.1 to 9.3], and Cx43(-/-) 1.1 [0.1 to 1.3; P<0.005]; in LV, WT 10.1 [9.4 to 10.7], Cx43(+/)(-) 8.3 [7.8 to 8.9], and Cx43(-/-) 1.7 [1.3 to 2.1; P<0.005]) corresponding with the downregulation of Cx40. Cx40 and Cx45 mRNAs were detectable in ventricular homogenates even at 17.5 dpc, probably accounting for the residual conduction function. Neonatal knockout hearts were arrhythmic in vivo as well as ex vivo. This study demonstrates the contribution of Cx43 to the electrical function of the developing mouse heart and the essential role of this gene in maintaining heart rhythm in postnatal life. PMID- 11397788 TI - Endothelial dysfunction and elevation of S-adenosylhomocysteine in cystathionine beta-synthase-deficient mice. AB - Hyperhomocysteinemia is associated with increased risk for cardiovascular events, but it is not certain whether it is a mediator of vascular dysfunction or a marker for another risk factor. Homocysteine levels are regulated by folate bioavailability and also by the methyl donor S-adenosylmethionine (SAM) and its metabolite S-adenosylhomocysteine (SAH). We tested the hypotheses that endothelial dysfunction occurs in hyperhomocysteinemic mice in the absence of folate deficiency and that levels of SAM and SAH are altered in mice with dysfunction. Heterozygous cystathionine beta-synthase-deficient (CBS(+/-)) and wild-type (CBS(+/+)) mice were fed a folate-replete, methionine-enriched diet. Plasma levels of total homocysteine were elevated in CBS(+/-) mice compared with CBS(+/+) mice after 7 weeks (27.1+/-5.2 versus 8.8+/-1.1 micromol/L; P<0.001) and 15 weeks (23.9+/-3.0 versus 13.0+/-2.3 micromol/L; P<0.01). After 15 weeks, but not 7 weeks, relaxation of aortic rings to acetylcholine was selectively impaired by 35% (P<0.05) and thrombomodulin anticoagulant activity was decreased by 20% (P<0.05) in CBS(+/-) mice. Plasma levels of folate did not differ between groups. Levels of SAH were elevated approximately 2-fold in liver and brain of CBS(+/-) mice, and correlations were observed between plasma total homocysteine and SAH in liver (r=0.54; P<0.001) and brain (r=0.67; P<0.001). These results indicate that endothelial dysfunction occurs in hyperhomocysteinemic mice even in the absence of folate deficiency. Endothelial dysfunction in CBS(+/-) mice was associated with increased tissue levels of SAH, which suggests that altered SAM-dependent methylation may contribute to vascular dysfunction in hyperhomocysteinemia. PMID- 11397789 TI - Elevated levels of S-nitrosoalbumin in preeclampsia plasma. AB - The availability of nitric oxide (NO), which is required for the normal regulation of vascular tone, may be decreased in preeclampsia, thus contributing to the vascular pathogenesis of this pregnancy disorder. Because ascorbate is essential for the decomposition of S-nitrothiols and the release of NO, we speculated that the ascorbate deficiency typical of preeclampsia plasma might result in decreased rates of decomposition of S-nitrosothiols. We tested the hypothesis that total S-nitrosothiol and S-nitrosoalbumin concentrations are increased in preeclampsia plasma, reflecting a decreased release of NO from these major reservoirs of NO. Gestationally matched plasma samples were obtained (before labor or intravenous MgSO(4)) from 21 women with preeclampsia and 21 women with normal pregnancy, and plasma samples were also obtained from 12 nonpregnant women of similar age and body mass index during the follicular phase of the menstrual cycle. All were nonsmokers. The assay included ultraviolet induced decomposition of S-nitrosothiols to liberate NO captured by a florigenic reagent, 4,5-diaminofluoresceine, to produce diaminofluoresceine-Triazole. Preeclampsia plasma contained significantly higher concentrations of total S nitrosothiols (11.1+/-2.9 nmol/mL) than normal pregnancy samples (9.4+/-1.5 nmol/mL). Even greater differences were found between preeclampsia plasma and plasma samples from normal pregnancies and nonpregnant women (294+/-110, 186+/ 25, and 151+/-25 pmol/mg protein, respectively) when S-nitrosothiol content was expressed per milligram protein. The albumin fraction contained 49.4% of total plasma S-nitrosothiols in the control samples and 53.7% and 56.8% of plasma S nitrosothiols in normal pregnancy and preeclampsia, respectively. The level of S nitrosoalbumin was significantly higher in preeclampsia than in normal pregnancy or nonpregnancy plasma (6.3+/-1.4, 5.1+/-0.7, and 4.2+/-1.0 nmol/mL, respectively). The increased concentration of S-nitrosoalbumin in preeclampsia almost completely accounted for the increased levels of S-nitrosothiols in total plasma. Due to combined increases in nitrosothiols and decreases in protein, the preeclampsia plasma concentration of S-nitrosoalbumin was greatly increased on a per milligram of protein basis (271% and 186% compared with normal nonpregnancy and normal pregnancy plasma, respectively). We conclude that S-nitrosoalbumin and total S-nitrosothiol concentrations are significantly increased in preeclampsia plasma and may reflect insufficient release of NO groups in this condition. PMID- 11397790 TI - Overwhelming evidence of the beneficial effects of SERCA gene transfer in heart failure. PMID- 11397791 TI - Phosphorylation of Thr(495) regulates Ca(2+)/calmodulin-dependent endothelial nitric oxide synthase activity. AB - The activity of the endothelial nitric oxide synthase (eNOS) can be regulated independently of an increase in Ca(2+) by the phosphorylation of Ser(1177) but results only in a low nitric oxide (NO) output. In the present study, we assessed whether the agonist-induced (Ca(2+)-dependent, high-output) activation of eNOS is associated with changes in the phosphorylation of Thr(495) in the calmodulin (CaM)-binding domain. eNOS Thr(495) was constitutively phosphorylated in porcine aortic endothelial cells and was rapidly dephosphorylated after bradykinin stimulation. In the same cells, bradykinin enhanced the phosphorylation of Ser(1177), which was maximal after 5 minutes, and abolished by the CaM-dependent kinase II (CaMKII) inhibitor KN-93. Bradykinin also enhanced the association of CaMKII with eNOS. Phosphorylation of Thr(495) was attenuated by the protein kinase C (PKC) inhibitor Ro 31-8220 and after PKC downregulation using phorbol 12 myristate 13-acetate. The agonist-induced dephosphorylation of Thr(495) was completely Ca(2+)-dependent and inhibited by the PP1 inhibitor calyculin A. Little CaM was bound to eNOS immunoprecipitated from unstimulated cells, but the agonist-induced dephosphorylation of Thr(495) enhanced the association of CaM. Mutation of Thr(495) to alanine increased CaM binding to eNOS in the absence of cell stimulation, whereas the corresponding Asp(495) mutant bound almost no CaM. Accordingly, NO production by the Ala(495) mutant was more sensitive to Ca(2+)/CaM than the aspartate mutant. These results suggest that the dual phosphorylation of Ser(1177) and Thr(495) determines the activity of eNOS in agonist-stimulated endothelial cells. Moreover, the dephosphorylation of Thr(495) by PP1 precedes the phosphorylation of Ser(1177) by CaMKII. The full text of this article is available at http://www.circresaha.org. PMID- 11397792 TI - Human mdm2 mediates multiple mono-ubiquitination of p53 by a mechanism requiring enzyme isomerization. AB - The mdm2 gene product is an important regulator of p53 function and stability. mdm2 is an E3 ubiquitin ligase for p53 and the RING finger domain of mdm2 is critical for ligase activity. Ubiquitin (Ub) conjugation is a general targeting modification and poly-ubiquitin chains specifically target proteins to the proteasome for degradation. In this report, we show that the multistep cascade of mdm2-mediated p53 ubiquitination can be reduced to three purified recombinant proteins: ubiquitin-conjugated E2, mdm2, and p53. This simplification allows enzymatic analysis of the isolated ligase reaction. The simplified reaction recapitulates the ubiquitination of p53 observed with individual components and the p53-Ub((n)) is qualitatively similar to p53-Ub((n)) detected in lactacystin treated cells. Surprisingly, we find that p53 is modified with multiple mono ubiquitin moieties as opposed to a poly-ubiquitin chain. Finally, kinetic analysis indicates the transfer reaction proceeds either through a modified Ping Pong mechanism involving requisite enzyme isomerization steps, or through a Rapid Equilibrium Random Bi Bi mechanism involving very large anti-cooperative interactions between the two substrate binding pockets on the enzyme, mediated through allosteric changes in enzyme structure. PMID- 11397793 TI - Identification and characterization of a new mammalian glutaredoxin (thioltransferase), Grx2. AB - A thiol/disulfide oxidoreductase component of the GSH system, glutaredoxin (Grx), is involved in the reduction of GSH-based mixed disulfides and participates in a variety of cellular redox pathways. A single cytosolic Grx (Grx1) was previously described in mammals. We now report identification and characterization of a second mammalian Grx, designated Grx2. Grx2 exhibited 36% identity with Grx1 and had a disulfide active center containing the Cys-Ser-Tyr-Cys motif. Grx2 was encoded in the genomes of mammals and birds and expressed in a variety of cell types. The gene for human Grx2 consisted of four exons and three introns, spanned 10 kilobase pairs, and localized to chromosome 1q31.2-31.3. The coding sequence was present in all exons, with the first exon encoding a mitochondrial signal peptide. The mitochondrial leader sequence was also present in mouse and rat Grx2 sequences and was shown to direct either Grx2 or green fluorescent protein to mitochondria. Alternative splicing forms of mammalian Grx2 mRNAs were identified that differed in sequences upstream of exon 2. To functionally characterize the new protein, human and mouse Grx2 proteins were expressed in Escherichia coli, and the purified proteins were shown to reduce mixed disulfides formed between GSH and S-sulfocysteine, hydroxyethyldisulfide, or cystine. Grx1 and Grx2 were sensitive to inactivation by iodoacetamide and H(2)O(2) and exhibited similar pH dependence of catalytic activity. However, H(2)O(2)-inactivated Grx2 could only be reactivated with 5 mm GSH, whereas Grx1 could also be reactivated with dithiothreitol or thioredoxin/thioredoxin reductase. The Grx2 structural model suggested a common reaction mechanism for this class of proteins. The data provide the first example of a mitochondrial Grx and also indicate the occurrence of a second functional Grx in mammals. PMID- 11397794 TI - Inhibition of AP-1 by the glucocorticoid-inducible protein GILZ. AB - The immunosuppressive effects of glucocorticoids arise largely by inhibition of cytokine gene expression, which has been ascribed to interference between the glucocorticoid receptor and transcription factors such as AP-1 and NF-kappa B as well as by competition for common coactivators. Here we show that glucocorticoid induced inhibition of interleukin-2 mRNA expression in activated normal T cells required new protein synthesis, suggesting that this phenomenon is secondary to expression of glucocorticoid-regulated genes. One of the most prominent glucocorticoid-induced genes is glucocorticoid-induced leucine zipper (GILZ), which has been reported to inhibit activation-induced up-regulation of Fas ligand (FasL) mRNA. Indeed, transient expression of GILZ in Jurkat T cells blocked induction of a reporter construct driven by the FasL promoter. This could be accounted for by GILZ-mediated inhibition of Egr-2 and Egr-3, NFAT/AP-1-inducible transcription factors that bind a regulatory element in the FasL promoter and up regulate FasL expression. GILZ also potently inhibited AP-1-driven and IL-2 promoter-driven reporter constructs, and recombinant GILZ specifically interacted with c-Fos and c-Jun in vitro and inhibited the binding of active AP-1 to its target DNA. Whereas homodimerization of GILZ required the presence of its leucine zipper, the interaction with c-Fos and c-Jun occurred through the N-terminal 60 amino acid region of GILZ. Thus, GILZ represents a glucocorticoid-induced gene product that can inhibit a variety of activation-induced events, at least in part by direct interference with AP-1, and is therefore a candidate for a mediator of glucocorticoid-induced immunosuppression. PMID- 11397795 TI - Constitutively dead, conditionally live HIV-1 genomes. Ex vivo implications for a live virus vaccine. AB - An effective vaccine against AIDS is unlikely to be available for many years. As we approach two decades since the first identification of human immunodeficiency virus, type 1 (HIV-1), currently, only one subunit vaccine candidate has reached phase 3 of clinical trials. The subunit approach has been criticized for its inability to elicit effectively cytotoxic T-lymphocyte (CTL) response, which is felt by many to be needed for protection against HIV-1 infection. In subhuman primates, a live attenuated simian immunodeficiency virus (SIV) vaccine candidate, capable of inducing CTL, has been found to confer prophylactic immunity sufficient to prevent simian AIDS. Because replication competent (live) attenuated viruses could over time revert to virulence, such a live attenuated approach has largely been dismissed for HIV-1. Here, we describe the creation of constitutively dead conditionally live (CDCL) HIV-1 genomes. These genomes are constitutively defective for the Tat/TAR axis and are conditionally dependent on tetracycline for attenuated replication with robust expression of viral antigens. Our results suggest that CDCL genomes merit consideration as safer "live" attenuated HIV-1 vaccine candidates. PMID- 11397796 TI - The amyloidogenic pathway of amyloid precursor protein (APP) is independent of its cleavage by caspases. AB - Amyloid beta-protein (A beta) is the main constituent of senile plaques in Alzheimer's disease and is derived by proteolysis from the amyloid precursor protein (APP). Generation and secretion of both A beta 40 and A beta 42 isoforms depend largely on internalization of APP and occurs mainly in the endocytic pathway. Evidence has also been presented (Gervais, F. G., Xu, D., Robertson, G. S., Vaillancourt, J. P., Zhu, Y., Huang, J., LeBlanc, A., Smith, D., Rigby, M., Shearman, M. S., Clarke, E. E., Zheng, H., Van der Ploeg, L. H. T., Ruffolo, S. C., Thornberry, N. A., Xanthoudakis, S., Zamboni, R. J., Roy, S., and Nicholson, D. W. (1999) Cell, 97, 395--406) that caspase cleavage of APP at its cytosolic tail affects its processing such that it is redirected to a more amyloidogenic pathway, resulting in enhanced A beta generation. However, caspase cleavage of APP also results in loss of its endocytosis signal (YENP), an event that would predict a decline in internalization and a concomitant decrease, not an increase, in A beta generation. In the present work, we examined whether caspase cleavage of APP is relevant to amyloidogenesis. We found that 1) caspase cleavage of APP results in reduced internalization and, accordingly, a decline in A beta secretion; 2) masking of the caspase site in APP did not affect A beta levels and, 3) caspase activation in cells by serum withdrawal did not increase A beta secretion. Thus, caspase cleavage of APP is unlikely to play a direct role in amyloidogenesis. PMID- 11397797 TI - The aggregation state of rhodanese during folding influences the ability of GroEL to assist reactivation. AB - The in vitro folding of rhodanese involves a competition between formation of properly folded enzyme and off-pathway inactive species. Co-solvents like glycerol or low temperature, e.g. refolding at 10 degrees C, successfully retard the off-pathway formation of large inactive aggregates, but the process does not yield 100% active enzyme. These data suggest that mis-folded species are formed from early folding intermediates. GroEL can capture early folding intermediates, and it loses the ability to capture and reactivate rhodanese if the enzyme is allowed first to spontaneously fold for longer times before it is presented to GroEL, a process that leads to the formation of unproductive intermediates. In addition, GroEL cannot reverse large aggregates once they are formed, but it could capture some folding intermediates and activate them, even though they are not capable of forming active enzyme if left to spontaneous refolding. The interaction between GroEL and rhodanese substantially but not completely inhibits intra-protein inactivation, which is responsible for incomplete activation during unassisted refolding. Thus, GroEL not only decreases aggregation, but it gives the highest reactivation of any method of assistance. The results are interpreted using a previously suggested model based on studies of the spontaneous folding of rhodanese (Gorovits, B. M., McGee, W. A., and Horowitz, P. M. (1998) Biochim. Biophys. Acta 1382, 120--128 and Panda, M., Gorovits, B. M., and Horowitz, P. M. (2000) J. Biol. Chem. 275, 63--70). PMID- 11397798 TI - Identification of a Delta 4 fatty acid desaturase from Thraustochytrium sp. involved in the biosynthesis of docosahexanoic acid by heterologous expression in Saccharomyces cerevisiae and Brassica juncea. AB - The existence of Delta 4 fatty acid desaturation in the biosynthesis of docosahexanoic acid (DHA) has been questioned over the years. In this report we describe the identification from Thraustochytrium sp. of two cDNAs, Fad4 and Fad5, coding for Delta 4 and Delta 5 fatty acid desaturases, respectively. The Delta 4 desaturase, when expressed in Saccharomyces cerevisiae, introduced a double bond at position 4 of 22:5(n-3) and 22:4(n-6) resulting in the production of DHA and docosapentanoic acid. The enzyme, when expressed in Brassica juncea under the control of a constitutive promoter, desaturated the exogenously supplied substrate 22:5(n-3), resulting in the production of DHA in vegetative tissues. These results support the notion that DHA can be synthesized via Delta 4 desaturation and suggest the possibility that DHA can be produced in oilseed crops on a large scale. PMID- 11397799 TI - Histamine-induced vasoconstriction involves phosphorylation of a specific inhibitor protein for myosin phosphatase by protein kinase C alpha and delta isoforms. AB - Histamine stimulus triggers inhibition of myosin phosphatase-enhanced phosphorylation of myosin and contraction of vascular smooth muscle. In response to histamine stimulation of intact femoral artery, a smooth muscle-specific protein called CPI-17 (for protein kinase C-potentiated inhibitory protein for heterotrimeric myosin light chain phosphatase of 17 kDa) is phosphorylated and converted to a potent inhibitor for myosin phosphatase. Phosphorylation of CPI-17 is diminished by pretreatment with either or GF109203x, suggesting involvement of multiple kinases (Kitazawa, T., Eto, M., Woodsome, T. P., and Brautigan, D. L. (2000) J. Biol. Chem. 275, 9897--9900). Here we purified and identified CPI-17 kinases endogenous to pig artery that phosphorylate CPI-17. DEAE-Toyopearl column chromatography of aorta extracts separated two CPI-17 kinases. One kinase was protein kinase C (PKC) alpha, and the second kinase was purified to homogeneity as a 45-kDa protein, and identified by sequencing as PKC delta. Purified PKC delta was 3-fold more reactive with CPI-17 compared with myelin basic protein, whereas purified PKC alpha and recombinant RhoA-activated kinases (Rho-associated coiled-coil forming protein Ser/Thr kinase and protein kinase N) showed equal activity with CPI-17 and myelin basic protein. inhibited CPI-17 phosphorylation by purified PKC delta with IC(50) of 0.6 microm (in the presence of 0.1 mm ATP) or 14 microm (2.0 mm ATP). significantly suppressed CPI-17 phosphorylation in smooth muscle cells, and the contraction of permeabilized rabbit femoral artery induced by stimulation with phorbol ester. GF109203x inhibited phorbol ester induced contraction of rabbit femoral artery by 80%, whereas a PKC alpha/beta inhibitor, Go6976, reduced contraction by 47%. The results imply that histamine stimulation elicits contraction of vascular smooth muscle through activation of PKC alpha and especially PKC delta to phosphorylate CPI-17. PMID- 11397800 TI - In vivo modifications of the maize mitochondrial small heat stress protein, HSP22. AB - A maize (Zea mays L.) small heat shock protein (HSP), HSP22, was previously shown to accumulate to high levels in mitochondria during heat stress. Here we have purified native HSP22 and resolved the protein into three peaks using reverse phase high performance liquid chromatography. Mass spectrometry (MS) of the first two peaks revealed the presence of two HSP22 forms in each peak which differed in mass by 80 daltons (Da), indicative of a monophosphorylation. Phosphorylation of HSP22 by [gamma-(32)P]ATP was also observed in mitochondria labeled in vitro, but not when purified native HSP22 was similarly used, demonstrating that HSP22 does not autophosphorylate, implicating a kinase involvement in vivo. Collisionally induced dissociation tandem MS (CID MS/MS) identified Ser(59) as the phosphorylated residue. We have also observed forms of HSP22 that result from alternative intron splicing. The two HSP22 proteins in the first peak were approximately 57 Da larger than the two HSP22 proteins in the second peak. MS analysis revealed that the +57-Da forms have an additional Gly residue directly N terminal of the expected Asp(84), which had been converted to an Asn residue. These results are the first demonstrations of phosphorylation and alternative intron splicing of a plant small HSP. PMID- 11397801 TI - An endoplasmic reticulum protein implicated in chaperoning peptides to major histocompatibility of class I is an aminopeptidase. AB - gp96, an abundant peptide-binding chaperone of the lumen of the endoplasmic reticulum and an acceptor of peptides transported into the endoplasmic reticulum through transporter associated with antigen processing, is shown to be an aminopeptidase. gp96 can trim an amino-terminal extended 19-mer precursor of the K(b)-binding VSV8 epitope for recognition by the cognate cytotoxic T lymphocyte clone. These observations support a role for gp96 in the amino-terminal trimming of extended peptides in the endoplasmic reticulum. PMID- 11397802 TI - The dimer initiation site hairpin mediates dimerization of the human immunodeficiency virus, type 2 RNA genome. AB - The untranslated leader of retroviral RNA genomes encodes multiple structural signals that are critical for virus replication. In the human immunodeficiency virus, type 1 (HIV-1) leader, a hairpin structure with a palindrome-containing loop is termed the dimer initiation site (DIS), because it triggers in vitro RNA dimerization through base pairing of the loop-exposed palindromes (kissing loops). Controversy remains regarding the region responsible for HIV-2 RNA dimerization. Different studies have suggested the involvement of the transactivation region, the primer binding site, and a hairpin structure that is the equivalent of the HIV-1 DIS hairpin. We have performed a detailed mutational analysis of the HIV-2 leader RNA, and we also used antisense oligonucleotides to probe the regions involved in dimerization. Our results unequivocally demonstrate that the DIS hairpin is the main determinant for HIV-2 RNA dimerization. The 6 mer palindrome sequence in the DIS loop is essential for dimer formation. Although the sequence can be replaced by other 6-mer palindromes, motifs that form more than two A/U base pairs do not dimerize efficiently. The inability to form stable kissing-loop complexes precludes formation of dimers with more extended base pairing. Structure probing of the DIS hairpin in the context of the complete HIV-2 leader RNA suggests a 5-base pair elongation of the DIS stem as it is proposed in current RNA secondary structure models. This structure is supported by phylogenetic analysis of leader RNA sequences from different viral isolates, indicating that RNA genome dimerization occurs by a similar mechanism for all members of the human and simian immunodeficiency viruses. PMID- 11397803 TI - Regulation of stearoyl coenzyme A desaturase expression in human retinal pigment epithelial cells by retinoic acid. AB - Stearoyl-CoA desaturase (SCD) is a regulatory enzyme involved in the synthesis of the monounsaturated fatty acids palmitoleate and oleate. The regulation of SCD is of physiological importance because the ratio of saturated fatty acids to unsaturated fatty acids is thought to modulate membrane fluidity. Differential display analysis of retinal pigment epithelial (ARPE-19) cells identified SCD as a gene regulated by retinoic acid. Two SCD transcripts of 3.9 and 5.2 kilobases in size were found to be expressed in these cells by Northern blot analysis. All trans-retinoic acid (all-trans-RA) increased SCD mRNA expression in a dose- and time-dependent manner; an approximately 7-fold increase was observed with 1 microm all-trans-RA at 48 h. SCD mRNA expression was also increased by 9-cis retinoic acid (9-cis-RA) as well as 4-(E-2-(5,6,7,8-tetrahydro-5,5,8,8 tetramethyl-2-naphthalenyl)-1-propenyl)benzoic acid (TTNPB), a retinoic acid receptor (RAR)-specific agonist. AGN194301, a RAR alpha-specific antagonist, suppressed the SCD expression induced by all-trans-RA, TTNPB, and 9-cis-RA. These results indicate the involvement of RAR alpha in the induction of SCD expression by retinoic acid. However, AGN194204, a RXR (retinoid X receptor) pan agonist, also increased SCD mRNA expression. This increase was not blocked by AGN194301, suggesting that an RAR-independent mechanism may also be involved. Thus, SCD expression in retinal pigment epithelial cells is regulated by retinoic acid, and the regulation appears to be mediated through RAR and RXR. PMID- 11397804 TI - Depolarization strongly induces human cytomegalovirus major immediate-early promoter/enhancer activity in neurons. AB - Activity-dependent changes in gene expression involving the transcription factor cAMP-response element-binding protein (CREB) occur in learning and memory, pain, and drug addiction. This mechanism may also be important for cytomegaloviral infections of the brain. The human cytomegalovirus major immediate-early promoter/enhancer (hCMV promoter), rate-limiting for productive cytomegalovirus infection, contains five cAMP-response elements (CREs). Indirect evidence suggests that this promoter does not function in unstimulated neurons. Here we test the hypothesis that expression from the hCMV promoter in neurons is induced by membrane depolarization. For these experiments, we infected cultured sympathetic and hippocampal neurons with hCMV-green fluorescent protein (GFP) promoter/reporter constructs using adenoviral gene transfer techniques and measured transgene expression by quantifying GFP fluorescence and GFP mRNA levels. We found that depolarization up-regulates promoter activity by >90-fold. Moreover, our results from pharmacological experiments suggest that this induction occurred through a CREB-dependent pathway. Importantly, site-directed mutagenesis of all five CREs in the promoter blocked this up-regulation almost completely, whereas mutating four of them had no effect. We conclude that the hCMV promoter acts as a molecular switch in neurons and is strongly induced by membrane depolarization, neuronal activity, or other stimuli that activate CREB. These results may provide insight into molecular mechanisms of cytomegalovirus related diseases of the brain. PMID- 11397805 TI - Alanine-scanning mutagenesis of the sixth transmembrane segment of gastric H+,K+ ATPase alpha-subunit. AB - The sixth transmembrane (M6) segment of the catalytic subunit plays an important role in the ion recognition and transport in the type II P-type ATPase families. In this study, we singly mutated all amino acid residues in the M6 segment of gastric H(+),K(+)-ATPase alpha-subunit with alanine, expressed the mutants in HEK 293 cells, and studied the effects of the mutation on the functions of H(+),K(+) ATPase; overall K(+)-stimulated ATPase, phosphorylation, and dephosphorylation. Four mutants, L819A, D826A, I827A, and L833A, completely lost the K(+)-ATPase activity. Mutant L819A was phosphorylated but hardly dephosphorylated in the presence of K(+), whereas mutants D826A, I827A, and L833A were not phosphorylated from ATP. We found that almost all of these amino acid residues, which are important for the function, are located on the same side of the alpha-helix of the M6 segment. In addition, we found that amino acids involved in the phosphorylation are located exclusively in the cytoplasmic half of the M6 segment and those involved in the K(+)-dependent dephosphorylation are in the luminal half. Several mutants such as I821A, L823A, T825A, and P829A partly retained the K(+)-ATPase activity accompanying the decrease in the rate of phosphorylation. PMID- 11397806 TI - Role of the T cell receptor ligand affinity in T cell activation by bacterial superantigens. AB - Similar to native peptide/MHC ligands, bacterial superantigens have been found to bind with low affinity to the T cell receptor (TCR). It has been hypothesized that low ligand affinity is required to allow optimal TCR signaling. To test this, we generated variants of Staphylococcus enterotoxin C3 (SEC3) with up to a 150-fold increase in TCR affinity. By stimulating T cells with SEC3 molecules immobilized onto plastic surfaces, we demonstrate that increasing the affinity of the SEC3/TCR interaction caused a proportional increase in the ability of SEC3 to activate T cells. Thus, the potency of the SEC3 variants correlated with enhanced binding without any optimum in the binding range covered by native TCR ligands. Comparable studies using anti-TCR antibodies of known affinity confirmed these observations. By comparing the biological potency of the two sets of ligands, we found a significant correlation between ligand affinity and ligand potency indicating that it is the density of receptor-ligand complexes in the T cell contact area that determines TCR signaling strength. PMID- 11397807 TI - Target genes of peroxisome proliferator-activated receptor gamma in colorectal cancer cells. AB - Activation of the nuclear hormone peroxisome proliferator-activated receptor gamma (PPARgamma) inhibits cell growth and promotes differentiation in a broad spectrum of epithelial derived tumor cell lines. Here we utilized microarray technology to identify PPARgamma gene targets in intestinal epithelial cells. For each gene, the induction or repression was seen with two structurally distinct PPARgamma agonists, and the change in expression could be blocked by co-treatment with a specific PPARgamma antagonist. A majority of the genes could be regulated independently by a retinoid X receptor specific agonist. Genes implicated in lipid transport or storage (adipophilin and liver fatty acid-binding protein) were also activated by agonists of PPAR subtypes alpha and/or delta. In contrast, PPARgamma-selective targets included genes linked to growth regulatory pathways (regenerating gene IA), colon epithelial cell maturation (GOB-4 and keratin 20), and immune modulation (neutrophil-gelatinase-associated lipocalin). Additionally, three different genes of the carcinoembryonic antigen family were induced by PPARgamma. Cultured cells treated with PPARgamma ligands demonstrated an increase in Ca(2+)-independent, carcinoembryonic antigen-dependent homotypic aggregation, suggesting a potential role for PPARgamma in regulating intercellular adhesion. Collectively, these results will help define the mechanisms by which PPARgamma regulates intestinal epithelial cell biology. PMID- 11397808 TI - Early intermediates in HIV-1 envelope glycoprotein-mediated fusion triggered by CD4 and co-receptor complexes. AB - An early step in the process of HIV-1 entry into target cells is the activation of its envelope glycoprotein (GP120-GP41) to a fusogenic state upon binding to target cell CD4 and cognate co-receptor. Incubation of human immunodeficiency virus (HIV)-1 Env-expressing cells with an excess of CD4 and co-recepeptor bearing target cells resulted in an influx of an impermeant nucleic acid-staining fluorescent dye into the Env-expressing cells. The dye influx occurred concomitant with cell fusion. No influx of dye into target cells was observed if they were incubated with an excess of Env-expressing cells. The permeabilization of Env-expressing cells was also triggered by CD4.co-receptor complexes attached to Protein G-Sepharose beads in the absence of target cells. The CD4 and co receptor-induced permeabilization of Env-expressing cells occurred with the same specificity with respect to co-receptor usage as cell fusion. Natural ligands for the co-receptors and C-terminal GP41 peptide inhibitors of HIV-1 fusion blocked this effect. Our results indicate that the process of HIV-1 Env-mediated fusion is initiated by the destabilization of HIV-1 Env-expressing membranes. Further elucidation of these early intermediates may help identify and develop potential inhibitors of HIV-1 entry into cells. PMID- 11397809 TI - IRAK1b, a novel alternative splice variant of interleukin-1 receptor-associated kinase (IRAK), mediates interleukin-1 signaling and has prolonged stability. AB - Interleukin-1 (IL-1) is a pleiotropic cytokine essential for initiation of the immune response to infections and stress. IL-1 interacts with its type I receptor (IL-1RI) and triggers a number of intracellular signaling cascades leading to activation of transcription factors, transcriptional up-regulation of target genes, and mRNA stabilization. IL-1RI-associated kinase-1 (IRAK1) is a membrane proximal serine-threonine kinase involved in IL-1 signaling that becomes phosphorylated and progressively degraded in response to IL-1 induction. We have identified a novel variant of IRAK1, which we have named IRAK1b, that arises from the use of an alternative 5'-acceptor splice site defined by sequence within exon 12 of IRAK1. IRAK1b mRNA exhibits wide tissue expression and is evolutionarily conserved in both mouse and human. IRAK1b can activate the transcription factor nuclear factor kappaB and interacts with the IL-1 signaling factors Toll interacting protein and tumor necrosis factor receptor-associated factor 6. It forms homodimers and heterodimers with the previously described isoform of IRAK1. We show that the IRAK1b protein is kinase-inactive and that, unlike IRAK1, its levels remain constant after IL-1 induction. The presence of an alternative splice variant of IRAK1, which is functionally active and highly stable following IL-1 stimulation, adds further complexity to the control mechanisms that govern IL-1 signaling. PMID- 11397810 TI - Mutational analysis of conserved residues in the GCN5 family of histone acetyltransferases. AB - GCN5 is a critical transcriptional co-activator and is the defining member of a large superfamily of N-acetyltransferases. GCN5 catalyzes the transfer of an acetyl group from acetyl-CoA to the epsilon-amino of lysine 14 within the core H3 histone protein. Previous biochemical analyses have indicated a fully ordered kinetic mechanism. Recent structural studies have implicated several conserved residues in catalysis and substrate binding. Here the roles of Glu-173, His-145, and Asp-214 in yeast GCN5 have been evaluated using site-directed mutagenesis, steady state and pre-steady state kinetics, pH analysis, isotope partitioning, and equilibrium binding studies. The results with wild type and E173Q, H145A, and D214A mutants are consistent with chemical catalysis being rate-determining in turnover. All mutants exhibited K(d) values (3.5-8.5 microm) for AcCoA that were similar to wild type enzyme, indicating no functional role for these residues in AcCoA binding. The E173Q mutant demonstrated a approximately 500-600-fold decreases in k(cat) and k(cat)/K(m),(H3), consistent with Glu-173 acting as the general base catalyst as proposed previously. No significant effect was observed on substrate binding steps. His-145 was identified as a residue in the peptide binding cleft that must be unprotonated (pK(a) = 5.8) for peptide binding and likely hydrogen-bonds to the Ser-10 hydroxyl of histone H3. His-145 also contributes to lowering the pK(a) value (by 0.8 units) of general base Glu-173 through a water-mediated hydrogen bond to the carboxylate side chain. Analysis of D214A revealed an obligate protein isomerization step that occurs after AcCoA binding and permits efficient peptide binding. Asp-214 is part of a conformationally flexible loop that mediates the isomerization by stabilizing distinct conformers of the protein. PMID- 11397811 TI - Conserved cysteine and tryptophan residues of the endothelin-converting enzyme-1 CXAW motif are critical for protein maturation and enzyme activity. AB - The neprilysin (NEP)/endothelin-converting enzyme (ECE) family of metalloproteases contains a highly conserved carboxyl-terminal tetrapeptide sequence, CXAW, where "C" is cysteine, "X" is a polar amino acid, "A" is an aliphatic residue, and "W" is tryptophan. Although this sequence strongly resembles a prenylation motif, human ECE-1 did not appear to be prenylated when labeled in vivo using various isoprenoid precursors in cell lines expressing ECE 1. We used site-directed mutagenesis to investigate the role of the CXAW motif and determined that the conserved cysteine residue of the CXAW motif in ECE-1, Cys(755), is critical for proper folding of the enzyme, its export from the endoplasmic reticulum, and its maturation in the secretory pathway. In addition, site-directed mutagenesis revealed that the conserved tryptophan residue of the sequence CEVW appears to be important for endoplasmic reticulum export and is essential for enzyme activity. Deletion of Trp(758) or substitution with alanine greatly slowed maturation of the enzyme, and resulted in more than a 90% loss of enzyme activity relative to the wild type. Conservative substitution of the tryptophan with phenylalanine did not reduce activity, whereas replacement with tyrosine, methionine, or leucine reduced enzyme activity by 50%, 75%, and 85%, respectively. Together, these data indicate that the conserved CEVW sequence does not serve as a prenylation signal and that both the conserved cysteine and tryptophan residues are necessary for proper folding and maturation of the enzyme. Furthermore, the conserved tryptophan appears to be critical for enzyme activity. PMID- 11397812 TI - C terminus-mediated control of voltage and cAMP gating of hyperpolarization activated cyclic nucleotide-gated channels. AB - The hyperpolarization-activated cyclic nucleotide-gated (HCN) family of "pacemaker" channels includes 4 isoforms, the kinetics and cAMP-induced modulation of which differ quantitatively. Because HCN isoforms are highly homologous in the central region, but diverge more substantially in the N and C termini, we asked whether these latter regions could contribute to the determination of channel properties. To this aim, we analyzed activation/deactivation kinetics and the response to cAMP of heterologously expressed isoforms mHCN1 and rbHCN4 and verified that mHCN1 has much faster kinetics and lower cAMP sensitivity than rbHCN4. We then constructed rbHCN4 chimeras by replacing either the N or the C terminus, or both, with the analogous domains from mHCN1. We found that: 1) replacement of the N terminus (chimera N1 4) did not substantially modify either the kinetics or cAMP dependence of wild type channels; 2) replacement of the C terminus, on the contrary, resulted in a chimeric channel (4-C1), the kinetics of which were strongly accelerated compared with rbHCN4, and that was fully insensitive to cAMP; 3) replacement of both N and C termini led to the same results as replacement of the C terminus alone. These results indicate that the C terminus of rbHCN4 contributes to the regulation of voltage- and cAMP-dependent channel gating, possibly through interaction with other intracellular regions not belonging to the N terminus. PMID- 11397813 TI - Oxygen access to the active site of cholesterol oxidase through a narrow channel is gated by an Arg-Glu pair. AB - Cholesterol oxidase is a monomeric flavoenzyme that catalyzes the oxidation and isomerization of cholesterol to cholest-4-en-3-one. Two forms of the enzyme are known, one containing the cofactor non-covalently bound to the protein and one in which the cofactor is covalently linked to a histidine residue. The x-ray structure of the enzyme from Brevibacterium sterolicum containing covalently bound FAD has been determined and refined to 1.7-A resolution. The active site consists of a cavity sealed off from the exterior of the protein. A model for the steroid substrate, cholesterol, can be positioned in the pocket revealing the structural factors that result in different substrate binding affinities between the two known forms of the enzyme. The structure suggests that Glu(475), located at the active site cavity, may act as the base for both the oxidation and the isomerization steps of the catalytic reaction. A water-filled channel extending toward the flavin moiety, inside the substrate-binding cavity, may act as the entry point for molecular oxygen for the oxidative half-reaction. An arginine and a glutamate residue at the active site, found in two conformations are proposed to control oxygen access to the cavity from the channel. These concerted side chain movements provide an explanation for the biphasic mode of reaction with dioxygen and the ping-pong kinetic mechanism exhibited by the enzyme. PMID- 11397814 TI - Characterization of the mammalian peroxisomal import machinery: Pex2p, Pex5p, Pex12p, and Pex14p are subunits of the same protein assembly. AB - Although many of the proteins involved in the biogenesis of the mammalian peroxisome have already been identified, our knowledge of the architecture of all this machinery is still very limited. In this work we used native gel electrophoresis and sucrose gradient sedimentation analysis in combination with immunoprecipitation experiments to address this issue. After solubilization of rat liver peroxisomes with the mild detergent digitonin, comigration of Pex5p, Pex14p, and a fraction of Pex12p was observed upon native electrophoresis and sucrose gradient sedimentation. The existence of a complex comprising Pex2p, Pex5p, Pex12p, and Pex14p was demonstrated by preparative coimmunoprecipitation experiments using an antibody directed to Pex14p. No stoichiometric amounts of Pex13p were detected in the Pex2p-Pex5p-Pex12p-Pex14p complex, although the presence of a small fraction of Pex13p in this complex could be demonstrated by Western blot analysis. Pex13p is also a component of a high molecular mass complex. Strikingly, partial purification of this Pex13p-containing complex revealed Pex13p as the major (if not the only) component. Taken together, our data indicate that Pex2p, Pex5p, Pex12p, and Pex14p, on one side, and Pex13p, on the other, are subunits of two stable protein complexes that probably interact with each other in the peroxisomal membrane. PMID- 11397815 TI - Activation of peroxisome proliferator-activated receptor gamma suppresses nuclear factor kappa B-mediated apoptosis induced by Helicobacter pylori in gastric epithelial cells. AB - Helicobacter pylori colonization leads to epithelial cell hyperproliferation within inflamed mucosa, but levels of apoptosis vary, suggesting that imbalances between rates of cell production and loss may contribute to differences in gastric cancer risk among infected populations. Peroxisome proliferator-activated receptor gamma (PPARgamma) regulates inflammatory and growth responses of intestinal epithelial cells. We determined whether activation of PPARgamma modified H. pylori-induced apoptosis in gastric epithelial cells. PPARgamma was expressed and functionally active in gastric epithelial cell lines sensitive to H. pylori-induced apoptosis. PPARgamma ligands 15d-PGJ(2) and BRL-49653 significantly attenuated H. pylomicronri-induced apoptosis, effects that could be reversed by co-treatment with a specific PPARgamma antagonist. Cyclopentanone prostaglandins that do not bind and activate PPARgamma had no effects on H. pylori-induced apoptosis. The ability of H. pylori to activate nuclear factor (NF)-kappaB and increase levels of the NF-kappaB target IL-8 was blocked by co treatment with PPARgamma agonists, and direct inhibition of NF-kappaB also abolished H. pylori-stimulated apoptosis. These results suggest that activation of the PPARgamma pathway attenuates the ability of H. pylori to induce NF-kappaB mediated apoptosis in gastric epithelial cells. Because PPARgamma regulates a multitude of host responses, activation of this receptor may contribute to varying levels of cellular turnover as well as the diverse pathologic outcomes associated with chronic H. pylori colonization. PMID- 11397816 TI - Involvement of Hgs/Hrs in signaling for cytokine-mediated c-fos induction through interaction with TAK1 and Pak1. AB - Hgs/Hrs is a tyrosine-phosphorylated FYVE finger protein that is induced by stimulation with various cytokines and growth factors. Here we show that Hgs plays critical roles in the signaling pathway for the interleukin-2-induced activation of the serum-response element and cyclic AMP-response element of the c fos promoter. We found that Hgs associated physically with transforming growth factor-beta-activated kinase 1 (TAK1) and p21-activated kinase 1 (Pak1), which mediate the activation of c-Jun N-terminal kinase and serum response factor, respectively, leading to transactivation via the serum-response element and cyclic AMP-response element. These results suggest that Hgs is involved in the TAK1-JNK and Pak1-serum response factor pathways for the c-fos induction that is initiated by cytokines. PMID- 11397817 TI - Affinity of human serum albumin for bilirubin varies with albumin concentration and buffer composition: results of a novel ultrafiltration method. AB - Albumin binding is a crucial determinant of bilirubin clearance in health and bilirubin toxicity in certain disease states. However, prior attempts to measure the affinity of albumin for bilirubin have yielded highly variable results, reflecting both differing conditions and the confounding influence of impurities. We therefore have devised a method based on serial ultrafiltration that successively removes impurities in [(14)C]bilirubin until a stable binding affinity is achieved, and then we used it to assess the effect of albumin concentration and buffer composition on binding. The apparent binding affinity of human serum albumin for [(14)C]bilirubin was strongly dependent on assay conditions, falling from (5.09 +/- 0.24) x 10(7) liters/mol at lower albumin concentrations (15 microm) to (0.54 +/- 0.05) x 10(7) liters/mol at higher albumin concentrations (300 microm). To determine whether radioactive impurities were responsible for this change, we estimated impurities in the stock bilirubin using a novel modeling approach and found them to be 0.11-0.13%. Formation of new impurities during the study and their affinity for albumin were also estimated. After correction for impurities, the binding affinity remained heavily dependent on the albumin concentration (range (5.37 +/- 0.26) x 10(7) liters/mol to (0.65 +/- 0.03) x 10(7) liters/mol). Affinities decreased by about half in the presence of chloride (50 mm). Thus, the affinity of human albumin for bilirubin is not constant, but varies with both albumin concentration and buffer composition. Binding may be considerably less avid at physiological albumin concentrations than previously believed. PMID- 11397819 TI - Reproductive function in human immunodeficiency virus infection. PMID- 11397820 TI - Calcium and bone metabolism in pregnancy and lactation. PMID- 11397821 TI - Hypothyroidism in pregnancy: consequences to neonatal health. PMID- 11397822 TI - The use of antithyroid drugs in pregnancy and lactation. PMID- 11397823 TI - Damaged reproduction: the most important consequence of iodine deficiency. PMID- 11397824 TI - Variation in the timing of puberty: clinical spectrum and genetic investigation. AB - Human puberty begins with the reemergence of GnRH secretion from its relative quiescence during childhood, activating a cascade of pituitary-gonadal maturation. This transition begins across a wide range of ages, and the rate of subsequent sexual maturation can be quite varied. The factors that regulate the hypothalamic-pituitary-gonadal axis and modulate the timing of puberty remain elusive, but it is clear that some regulation is under genetic control. Here, we discuss how new advances in genetic research may provide the tools to help unravel this long-standing mystery. PMID- 11397825 TI - Effects of age on testicular function and consequences of testosterone treatment. PMID- 11397826 TI - Reproductive technologies for male infertility. PMID- 11397827 TI - Androgen replacement therapy in the aging male--a critical evaluation. PMID- 11397828 TI - Erectile dysfunction. PMID- 11397829 TI - Androgen deficiency in women. AB - Physiological and pathological processes as well as iatrogenic interventions may result in androgen deficiency compared with levels in young healthy women. Whether relative androgen deficiency results in a clinical syndrome similar to that reported in men, including osteopenia, increased fat mass, decreased libido, and diminished quality of life, has not been definitively established. However, preliminary data in postmenopausal women suggest that physiological androgen replacement therapy, which involves substantially lower doses than those used in men, may result in increased bone mineral density, increased libido, and improved quality of life. The safety of androgen preparations that result in supraphysiological levels has not been established in women and would be expected to result in hirsutism, acne, and virilization with chronic use. Androgen preparations that avoid liver metabolism and result in physiological serum androgen levels in women with androgen deficiency are not currently available, but are in development. Therefore, although widespread screening and hormone replacement for androgen deficiency cannot be recommended yet, increasing interest in this topic makes consideration of the available data important. PMID- 11397831 TI - Estrogen modulates the hypothalamic-pituitary-adrenal and inflammatory cytokine responses to endotoxin in women. AB - Endotoxin stimulates the release of the inflammatory cytokines interleukin (IL) 1, IL-6, and tumor necrosis factor (TNF)-alpha, which are potent activators of the hypothalamic-pituitary-adrenal (HPA) axis. Recent studies in the rodent and in the primate have shown that the HPA responses to endotoxin and IL-1 were enhanced by gonadectomy and attenuated by estradiol (E2) replacement. In addition, there is some evidence, in the rodent, that estrogen modulates inflammatory cytokine responses to endotoxin. To determine whether estrogen has similar effects in humans, we studied the cytokine and HPA responses to a low dose of endotoxin (2--3 ng/kg) in six postmenopausal women with and without transdermal E2 (0.1 mg) replacement. Mean E2 levels were 7.3 +/- 0.8 pg/mL in the unreplaced subjects and increased to 102 +/- 13 pg/mL after estrogen replacement. Blood was sampled every 20 min for 1--2 h before, and for 7 h after, iv endotoxin administration. Endotoxin stimulated ACTH, cortisol, and cytokine release in women with and without E2 replacement. E2 significantly attenuated the release of ACTH (P < 0.0001) and of cortisol (P = 0.02). Mean ACTH levels peaked at 190 +/- 91 pg/mL in the E2-replaced group vs. 411 +/- 144 pg/mL in the unreplaced women, whereas the corresponding mean cortisol levels peaked at 27 +/- 2.9 microg/dL with E2 vs. 31 +/- 3.2 microg/dL without E2. Estrogen also attenuated the endotoxin-induced release of IL-6 (P = 0.02), IL-1 receptor antagonist (P = 0.003), and TNF-alpha (P = 0.04). Mean cytokine levels with and without E2 replacement peaked at 341 +/- 94 pg/mL vs. 936 +/- 620 pg/mL for IL-6, 82 +/- 14 ng/mL vs. 133 +/- 24 ng/mL for IL-1 receptor antagonist, and 77 +/- 46 pg/mL vs. 214 +/- 87 pg/mL for TNF-alpha, respectively. We conclude that inflammatory cytokine and HPA responses to a low dose of endotoxin are attenuated in postmenopausal women receiving E2 replacement. These data show, for the first time in the human, that a physiological dose of estrogen can restrain cytokine and neuroendocrine responses to an inflammatory challenge. PMID- 11397832 TI - Prenatal leptin production: evidence that fetal adipose tissue produces leptin. AB - In the adult, circulating leptin is highly correlated to adipose tissue mass. Whether such a relationship exists prenatally is unknown, because the actual source of fetal leptin has not been determined. In the present study, we have assessed the placental contribution to fetal and maternal circulating leptin concentrations and determined whether fetal adipose tissue produces leptin. The rate of leptin production in dually perfused human placenta was 0.036 ng/min.g. Ninety-five percent of the leptin released was delivered into the maternal circulation, vs. only 5% on the fetal side. Leptin messenger RNA and protein were detected in adipose tissue biopsies of 20-38 week human fetuses. However, leptin concentration was twice lower in fetal (0.22 +/- 0.11 ng/mg protein, n = 6) than in adult (0.49 +/- 0.12 ng/mg protein, n = 8) adipose tissue. Umbilical leptin levels closely reflected ponderal index at birth over a wide range of birth weights (1.6--4.1 kg). In sharp contrast, maternal and placental leptin concentrations were increased in pregnancies associated with fetal growth retardation. We conclude that umbilical leptin levels are independent of placental leptin production and can be taken as a marker of fat mass in human fetuses. By contrast, placental leptin production makes a substantial contribution to maternal circulating leptin levels during pregnancy. PMID- 11397833 TI - Sertoli cell function in infertile patients with and without microdeletions of the azoospermia factors on the Y chromosome long arm. AB - Deletions of the azoospermia factors on the Y chromosome long arm are an important cause of male infertility, and they may involve germ cell-specific genes or ubiquitously expressed genes. To date, no clinical or hormonal parameters have yet been found to distinguish patients with and without Yq microdeletions. In particular, Sertoli cell function, as evaluated by inhibin B, has not yet been described. Our hypothesis was that microdeletions involving genes specifically expressed in germ cells should not alter Sertoli cell function. To do this, we have evaluated the testicular hormonal function in infertile patients affected by severe testiculopathies with and without Yq microdeletions, with particular emphasis on Sertoli cell function. We studied 102 well-characterized infertile patients; 27 had Yq microdeletions, and 75 were classified as idiopathic infertiles. Patients with Yq microdeletions had lower FSH and higher inhibin B plasma concentrations with respect to patients without microdeletions, suggesting that Sertoli cell function in Yq-deleted men is only partially altered. Furthermore, patients with deletions involving germ cell specific genes had higher concentrations of inhibin B with respect to patients with deletions of ubiquitously expressed genes. These results suggested that a specific alteration of germ cells only partially influences Sertoli cell function. Hormonal status of patients without deletions suggested that in such cases the cause that has determined the spermatogenic defect may have damaged both Sertoli and germ cells. Inhibin B production in patients with Yq deletions was about 70% higher than the nondeleted patients, and the functional relationship between FSH and inhibin B was normally preserved. This study elucidated the multifactorial mechanisms underlying spermatogenic defects, where Sertoli cells may be normally functioning or damaged depending on the primary cause that has determined the testicular damage. PMID- 11397834 TI - Messenger ribonucleic acid encoding interferon-inducible guanylate binding protein 1 is induced in human endometrium within the putative window of implantation. AB - The putative window of embryo implantation in the human opens between days 19--24 of the menstrual cycle. During this period, the endometrium undergoes distinctive structural and functional changes orchestrated by steroid hormones, growth factors, and cytokines to attain a receptive phase in which it acquires the ability to implant the developing embryo. A major challenge in the study of human reproduction is to identify the molecular signals that participate in the establishment of this critical receptive phase in the context of the natural cycle. Toward this goal, we analyzed human endometrial biopsies at various days of the menstrual cycle by employing messenger RNA (mRNA) differential display technique. We isolated several complementary DNAs representing genes that are either up- or down-regulated within the putative window of implantation. We identified one of these genes as that encoding interferon (IFN)-inducible guanylate-binding protein 1 (or GBP1), which possesses GTPase activity. Analysis of endometrial biopsies by Northern blotting and RT-PCR demonstrated that GBP1 mRNA is specifically induced at the midsecretory phase of the menstrual cycle. In situ hybridization analysis revealed that GBP1 mRNA expression is localized in the glandular epithelial cells as well as in the stroma in the immediate vicinity of the glands. We observed that treatment of human endometrial adenocarcinoma cell, Ishikawa, with IFN-gamma or IFN-alpha markedly induced the expression of GBP1 mRNA. IFN-gamma was, however, a more potent inducer of GBP1 than IFN-alpha. Consistent with this finding, the temporal profile of GBP1 expression during the menstrual cycle resembled that of IFN-gamma mRNA more closely than that of IFN alpha, predicting a regulatory role of IFN-gamma in GBP1 expression in midsecretory human endometrium. Although the precise function of GBP1 in the receptive human uterus remains unclear, its unique expression overlapping the putative window of implantation suggests that it might serve as a useful marker of uterine receptivity in the human. PMID- 11397835 TI - Specific factors predict the response to pulsatile gonadotropin-releasing hormone therapy in polycystic ovarian syndrome. AB - Ovulation induction is particularly challenging in patients with polycystic ovarian syndrome (PCOS) and may be complicated by multifollicular development. Pulsatile GnRH stimulates monofollicular development in women with anovulatory infertility; however, ovulation rates are considerably lower in the subgroup of patients with PCOS. The aim of this retrospective study was to determine specific hormonal, metabolic, and ovarian morphological characteristics that predict an ovulatory response to pulsatile GnRH therapy in patients with PCOS. Subjects with PCOS were defined by chronic amenorrhea or oligomenorrhea and clinical and/or biochemical hyperandrogenism in the absence of an adrenal or pituitary disorder. At baseline, gonadotropin dynamics were assessed by 10-min blood sampling, insulin resistance by fasting insulin levels, ovarian morphology by transvaginal ultrasound, and androgen production by total testosterone levels. Intravenous pulsatile GnRH was then administered. During GnRH stimulation, daily blood samples were analyzed for gonadotropins, estradiol (E(2)), progesterone, inhibin B, and androgen levels, and serial ultrasounds were performed. Forty-one women with PCOS underwent a total of 144 ovulation induction cycles with pulsatile GnRH. Fifty-six percent of patients ovulated with 40% of ovulatory patients achieving pregnancy. Among the baseline characteristics, ovulatory cycles were associated with lower body mass index (P < 0.05), lower fasting insulin (P = 0.02), lower 17-hydroxyprogesterone and testosterone responses to hCG (P < 0.03) and higher FSH (P < 0.05). In the first week of pulsatile GnRH treatment, E(2) and the size of the largest follicle were higher (P < 0.03), whereas androstenedione was lower (P < 0.01) in ovulatory compared with anovulatory patients. Estradiol levels of 230 pg/mL (844 pmol/L) or more and androstenedione levels of 2.5 ng/mL (8.7 nmol/L) or less on day 4 and follicle diameter of 11 mm or more by day 7 of pulsatile GnRH treatment had positive predictive values for ovulation of 86.4%, 88.4%, and 99.6%, respectively. Ovulatory patients who conceived had lower free testosterone levels at baseline (P < 0.04). In conclusion, pulsatile GnRH is an effective and safe method of ovulation induction in a subset of patients with PCOS. Patient characteristics associated with successful ovulation in response to pulsatile GnRH include lower body mass index and fasting insulin levels, lower androgen response to hCG, and higher baseline FSH. In ovulatory patients, high free testosterone is negatively associated with pregnancy. A trial of pulsatile GnRH therapy may be useful in all PCOS patients, as E(2) and androstenedione levels on day 4 or follicle diameter on day 7 of therapy are highly predictive of the ovulatory response in this group of patients. PMID- 11397836 TI - Pharmacokinetics of a transdermal testosterone system in men with end stage renal disease receiving maintenance hemodialysis and healthy hypogonadal men. AB - Androgen deficiency is common in men with end stage renal disease (ESRD) on maintenance hemodialysis. Pharmacokinetics of transdermal testosterone in men receiving maintenance hemodialysis have not been studied. Our objective was to compare the pharmacokinetics of a transdermal testosterone system in healthy hypogonadal men and in men with ESRD on maintenance hemodialysis. We recruited 10 healthy hypogonadal men and 8 medically stable men on maintenance hemodialysis, 18--70 yr old, who had serum testosterone less than 300 ng/dL. After baseline sampling during a 24-h control period, two testosterone patches were applied daily for 28 days, to achieve a nominal delivery of 10-mg testosterone daily. In addition to single, pooled samples on days 7, 14, and 21, blood was drawn at 0, 2, 4, 6, 8, and 24 h on day 28 in healthy hypogonadal men and on an interdialytic day (day 21 or 28) as well as a dialysis day (day 21 or 28) in men on hemodialysis. On the dialysis day (day 21 or 28), serum free and total testosterone levels were measured hourly for 4 h before hemodialysis and for 4 h during hemodialysis. The dialysate was sampled for testosterone measurement. Baseline mean + SD total (92 +/- 82 vs. 222 +/- 50 ng/dL) and free (11 +/- 9 vs. 27 +/- 6 pg/mL) testosterone concentrations were lower in healthy hypogonadal men than in men with ESRD. After application of two testosterone patches, serum total and free testosterone concentrations rose into the midnormal range in both groups of men. Time-average, steady state (total testosterone, 506 +/- 88 vs. 516 +/- 86 ng/dL; free testosterone, 55 +/- 9 vs. 67 +/- 11 pg/mL), minimum, and maximum total and free testosterone concentrations were not significantly different between the two groups of men during treatment. Increments in total and free testosterone concentrations above baseline, baseline-subtracted areas under the total and free testosterone curves, and half-life of testosterone elimination (t(1/2), 2.1 +/- 0.1 vs. 2.1 +/- 0.2 h, P = not significant) were not significantly different between the two groups. In men receiving hemodialysis, time-average, steady state, and maximal total and free testosterone concentrations and baseline-subtracted areas under the total and free testosterone curves were higher on dialysis day than on an interdialytic day. On the day of hemodialysis, time-average total and free testosterone concentrations were not significantly different during the 4 h before or during hemodialysis. The amount of testosterone removed in the dialysate (8.4 +/- 1.6 microg during 4 h of hemodialysis) was small compared with the daily testosterone production rates in healthy young men. Serum dihydrotestosterone and estradiol concentrations increased into the normal male range and were not significantly different between the two groups. Percent suppression of LH was greater in men with ESRD than in healthy hypogonadal men. A regimen of two Testoderm TTS testosterone patches (Alza Corp., Mountain View, CA) daily can maintain serum concentrations of total and free testosterone and its metabolites dihydrotestosterone and estradiol in the midnormal range in healthy hypogonadal men and men on hemodialysis. The amount of testosterone cleared by hemodialysis is small, and hemodialysis does not significantly affect serum total and free testosterone concentrations in men treated with the testosterone patch. PMID- 11397837 TI - Gonadal status in male survivors following childhood brain tumors. AB - The effect of radiotherapy (RT) and chemotherapy (CT) on gonadal function was assessed in males treated for a childhood brain tumor not directly involving the hypothalamus/pituitary (HP) axis in a population-based study with a long follow up time. All males <15 yr at the time of diagnosis (median: 9.0 yr, range: 0.8 to 14.9 yr) and diagnosed from January 1970 through February 1997 in the eastern part of Denmark and [gte]18 yr at the time of follow-up (median: 25.8 yr, range:18.5 to 39.3 yr) were included. Thirty males fulfilled the criteria. The median age at time of RT was 9.0 yr (range: 0.8 to 14.9 yr) and the median length of follow-up was 18 yr (range: 2.0 to 28.0 yr). The biological effective dose of RT was determined to the HP region and to the spine and expressed in gray because the biological effective dose gives a means of expressing the biological effect on normal tissue of different dosage schedules in a uniform way. Levels of serum FSH, luteinizing hormone (LH), sexual hormone-binding globulin, testosterone, and inhibin B were measured and compared with healthy age-matched male controls (n = 347), and the patients had a GnRH stimulation test performed with determination of peak FSH and LH. Patients treated with RT + CT (n = 13), compared with patients treated with RT only (n = 17), had significantly higher median peak FSH (8.33 vs. 3.79 IU/L, P = 0.03) and median peak LH (20.0 vs. 12.8 IU/L, P = 0.03), and significantly lower median inhibin B (86.0 vs. 270 pg/ml, P = 0.03), and median inhibin B/FSH ratio (12.8 vs. 107.9, P = 0.04), which indicates gonadal damage. Inhibin B and inhibin B/FSH ratio were also significantly lower in the RT + CT group, compared with controls (median: 86.0 vs. 215 pg/ml, P = 0.02), (median:12.8 vs. 67; P = 0.01), respectively. We found a significantly inverse correlation between basal FSH and inhibin B and FSH and total testicular volume (r(s) = -0.83; P < 0.0001), (r(s) = -0.67; P < 0.0001), respectively, and a significant correlation between inhibin B and total testicular volume (r(s) = 0.63; P < 0.0001). Stepwise backward multiple linear regression analysis showed the best-fit model to predict inhibin B levels included total testicular volume (P = 0.002) and CT (P = 0.09). Median basal LH in the RT-only group was significantly lower, compared with controls (3.44 vs. 2.45 IU/L; P = 0.0001) indicating secondary hypogonadism, and in both the RT + CT group and the RT-only group, levels of testosterone were significantly lower, compared with our reference population (12.8 vs. 21.9 nmol/L; P = 0.001, and 14.7 vs. 21.9 nmol/L; P = 0.0003), respectively. In conclusion these data suggest that cranial irradiation for a childhood brain tumor may affect the HP axis, and adjuvant CT can reduce inhibin B indicating primary gonadal damage. Thus, such patients may have normal or even low levels of FSH despite damage to the seminiferous epithelium, and because the fertility status by a semen analysis for psychological reasons can be difficult to obtain in this group of patients, we suggest inhibin B as the most useful direct serum marker of spermatogenesis in the follow-up of individuals who have received both cranial irradiation and gonadotoxic chemotherapy. However, because the number of patients with RT + CT and RT only are small, these data must be confirmed in further studies. PMID- 11397838 TI - Low grade chronic inflammation in women with polycystic ovarian syndrome. AB - Low grade chronic inflammation as reflected by increased C-reactive protein (CRP) concentrations independently predicts those at risk for coronary heart disease (CHD) and type 2 diabetes. Women with polycystic ovarian syndrome (PCOS) are insulin resistant and have increased risk for CHD and type 2 diabetes, but currently there are no data on markers of inflammation in women with PCOS. Seventeen women with PCOS (defined on the basis of elevated testosterone and oligomenorrhea) and 15 healthy women matched as a group for body mass index were recruited. Measurement of CRP concentrations was made using a highly sensitive assay. Insulin resistance was assessed using the hyperinsulinemic euglycemic clamp technique. The women with PCOS had significantly elevated CRP concentrations relative to controls (geometric means, 2.12 and 0.67 mg/L, respectively; P = 0.016). Log CRP correlated with body mass index in both PCOS and controls (r = 0.58; P < 0.05 and r = 0.78; P < 0.01, respectively) and inversely with insulin sensitivity (r = -0.57; P < 0.05 and r = -0.69; P < 0.01). Total testosterone did not correlate with log CRP in either group. On adjustment for body mass index and age, there remained a significant difference in log CRP between PCOS and controls (t = 2.13; P < 0.05). On further adjustment for insulin sensitivity, log CRP was no longer significantly different between groups (t = 1.51; P = 0.14). We conclude that women with PCOS have significantly increased CRP concentrations relative to women with normal menstrual rhythm and normal androgen levels. We propose low grade chronic inflammation as a novel mechanism contributing to increased risk of CHD and type 2 diabetes in these women. PMID- 11397839 TI - McCune-Albright syndrome: growth hormone dynamics in pregnancy. AB - Excess GH secretion has a well recognized association with McCune-Albright syndrome. Although there have been a number of reported pregnancies in uncontrolled acromegaly, none has been described in the McCune-Albright syndrome. We have studied the GH and insulin-like growth factor I (IGF-I) profiles in a patient with confirmed McCune-Albright syndrome and GH hypersecretion throughout a successful pregnancy and postpartum period. Prepregnancy, IGF-I was 60.6 nmol/L (normal, 18.0--43.1), and the daytime GH profile measured using assay A was 9.6- 14.0 mU/L. At 13 weeks gestation there was a decline of IGF-I to 33.9 nmol/L and in the daytime GH profile (assay A) to 5.4--6.8 mU/L. At 24 weeks, IGF-I had risen to 51.6 nmol/L. A simultaneous daytime GH profile at this time using assay A revealed levels between 21.3--22.1 mU/L, but only 2.1--3.0 mU/L with assay B. Assay A has significant cross-reactivity with human placental lactogen (HPL), unlike assay B. At 36 weeks, IGF-I was still elevated at 56.6 nmol/L, with a daytime GH profile of 16.6--17.7 mU/L using assay A and 1.5--3.9 mU/L with assay B. At 12 weeks postpartum, IGF-I was 71.4 nmol/L, and the daytime GH profile with assay B was 5.6--8.6 mU/L. These data support a picture of GH suppression during pregnancy in acromegaly associated with McCune-Albright syndrome, shown best with assay B, which discriminates between GH and HPL. These results contrast with previous reports of pregnancy in uncontrolled acromegalics, in whom pituitary GH levels were unaffected by pregnancy, and total GH and IGF-I levels were noted to be elevated. These data suggest that GH secretion in a pregnant acromegalic with the McCune-Albright syndrome may not be entirely autonomous, as seen in classic acromegaly, but may be associated with a degree of negative feedback control that could be exerted by a circulating factor of placental origin, probably HPL or placental GH. PMID- 11397840 TI - Growth hormone (GH) secretion in patients with an inactivating defect of the GH releasing hormone (GHRH) receptor is pulsatile: evidence for a role for non-GHRH inputs into the generation of GH pulses. AB - GH secretion is regulated by the interaction of GHRH and somatostatin and is released in 10-20 pulses in each 24-h cycle. The exact roles in pulse generation played by somatostatin, GHRH, and the recently isolated GH-releasing peptide, Ghrelin, are not fully elucidated. To investigate the GHRH-mediated GH secretion in human, we investigated pulsatile, entropic, and 24-h rhythmic GH secretion in two young adults (male, 24 yr; female, 23 yr) from a Moroccan family with a novel inactivating defect of the GHRH receptor gene. Data were compared with values in age- and gender-matched controls. Plasma GH concentration were measured by a sensitive immunofluorometric assay, with a detection limit of 0.01 mU/L. All plasma GH concentrations in the female patient were measurable; in the male patient 30 of 145 samples were at or below the detection limit. GH secretion was pulsatile, with 21 and 23 secretory episodes/24 h in the male and female patients, respectively. The fraction of basal to total GH secretion was raised in both patients by 0.18 and 0.15, respectively. The total 24-h GH production rate was greatly diminished; in the male patient it was 6.9 mU/L (normal values for his age, 26--63 mU/L), and in the female patient it was 4.2 mU/L (normal values for her age, 96--390 mU/L). The nyctohemeral plasma GH rhythm was preserved (P < 0.001), with normal acrophases (0430 and 0218 h in the male and female, respectively). Approximate entropy was greatly elevated in both subjects (0.82 in the male and 1.17 in the female; upper normal values for age and gender, 0.24 and 0.59, respectively). Intravenous injection of 50 microg GHRH failed to increase the plasma GH concentration in both patients, but 100 microg GH-releasing peptide 2 elicited a definite increase (male patient, 0.13 to 1.74 mU/L; female patient, 0.29 to 0.87 mU/L). Both patients had a partial empty sella on magnetic resonance imaging scanning. In summary, the present studies in two patients with a profound loss of function mutation of the GHRH receptor favor the view that in the human the timing of GH pulses is primarily supervised by intermittent somatostatin withdrawal, and the amplitude of GH pulses is driven by GHRH. In addition, we infer that effectual GHRH input controls the GH cell mass and the orderliness of the secretory process. PMID- 11397841 TI - Absence of mutations involving the LIM homeobox domain gene LHX9 in 46,XY gonadal agenesis and dysgenesis. AB - The etiology of most cases of 46,XY gonadal dysgenesis in the absence of extragenital anomalies is not accounted for by mutations in the genes known to date to be involved in sex determination. We have investigated the possibility that mutations in the gene LHX9, whose murine ortholog causes isolated gonadal agenesis when inactivated, might be responsible for gonadal dysgenesis and agenesis in humans. We isolated a human LHX9 complementary DNA (cDNA), mapped the gene to the long arm of human chromosome 1, and determined its genomic structure. We found that LHX9 is highly conserved between species, sharing in particular over 98% amino acid identity. A mutational screen was performed in a sample of patients with a range of gonadal maldevelopment, including bilateral gonadal agenesis in two sisters with an opposite sex karyotype. We did not detect mutations in the open reading frame of LHX9 in the patients studied. However, the extent of between-species structural conservation suggests that LHX9 deserves further consideration as a determinant of gonadal function in humans. PMID- 11397842 TI - The fertile eunuch variant of idiopathic hypogonadotropic hypogonadism: spontaneous reversal associated with a homozygous mutation in the gonadotropin releasing hormone receptor. AB - Mutations in the GnRH receptor (GnRH-R) gene have been reported to cause idiopathic hypogonadotropic hypogonadism (IHH). Herein, we describe a 26-yr-old male with a mild phenotypic form of IHH, the fertile eunuch syndrome (IHH in the presence of normal testicular size and some degree of spermatogenesis), associated with a homozygous mutation (Gln106Arg) in the GnRH-R. This mutation, located in the first extracellular loop of the GnRH-R, has been previously shown to decrease but not eliminate GnRH binding. The proband had hypogonadal testosterone levels, detectable but apulsatile gonadotropin secretion, and a normal adult male testicular size of 17 mL at baseline. After only 4 months of treatment with hCG alone, he developed sperm in his ejaculate and his wife conceived. Following cessation of hCG therapy, the patient demonstrated reversal of his hypogonadotropism as evidenced by normal adult male testosterone levels and the appearance of pulsatile luteinizing hormone secretion. This case thus expands the emerging clinical spectrum of GnRH-R mutations, provides the first genetic basis for the fertile eunuch variant of IHH and documents the occurrence of reversible IHH in a patient with a GnRH-R mutation. PMID- 11397843 TI - The relationship between steroidogenic factor 1 and DAX-1 expression and in vitro gonadotropin secretion in human pituitary adenomas. AB - The orphan nuclear receptors, steroidogenic factor 1 (SF-1) and DAX-1, are involved in gonadotroph differentiation, and SF-1 has been shown to activate the LH-beta and glycoprotein hormone alpha-subunit (alpha GSU) gene promoters. Pituitary adenomas from 34 patients [13 somatotroph tumors, 4 prolactinomas, and 17 clinically nonfunctioning pituitary adenomas (NFPAs)] were enzymatically dispersed and cultured in vitro for 48 h. Tissue culture medium was collected and assayed for LH, FSH, and alpha GSU; messenger RNA was extracted from adherent cells, and expression of SF-1 and DAX-1 messenger RNA was determined by RT-PCR and verified by direct DNA sequencing. The presence of DAX-1 protein in tumor tissue was confirmed by immunocytochemistry. DAX-1 was demonstrated in all NFPAs, 7 of 13 somatotroph tumors and 0 of 4 prolactinomas. SF-1 expression occurred in 8 of 16 NFPAs, 4 of 12 somatotroph tumors, and 1 of 4 prolactinomas. LH secretion in vitro was greater in NFPAs that were SF-1 positive (P < 0.05). Neither FSH secretion nor alpha GSU secretion in vitro were significantly related to the expression of SF-1 or DAX-1. SF-1-positive somatotroph tumors immunostained positively for LH-beta and/or FSH-beta and secreted gonadotropins in vitro. SF-1 expression is associated with the in vitro secretion of LH by NFPAs. A proportion of somatotroph tumors also express SF-1 and DAX-1 and secrete gonadotropin hormones in vitro. PMID- 11397844 TI - Insulin-like growth factor-binding protein 1 stimulates human trophoblast migration by signaling through alpha 5 beta 1 integrin via mitogen-activated protein Kinase pathway. AB - A highly migratory subpopulation of the human placental trophoblast, known as the extravillous trophoblast (EVT), invades the uterus and its vasculature, to establish adequate exchange of key molecules between the maternal and fetal circulations. During their formation, EVT cells selectively acquire alpha 5 beta 1 integrin. We had shown that alpha 5 beta 1 is required for their migratory function, and that EVT cell migration is stimulated by insulin-like growth factor binding protein (IGFBP)-1 produced by the uterine decidua. The present study examined whether this stimulation is dependent on binding of the Arg-Gly-Asp (RGD) domain of IGFBP-1 to an RGD binding site on the alpha 5 beta 1 integrin, followed by activation of focal adhesion kinase (FAK) and stimulation of the mitogen-activated protein kinase (MAPK) pathway. IGFBP-1 treatment increased migration of EVT cells, whereas an anti-alpha 5 beta 1 integrin antibody blocked migration regardless of IGFBP-1 treatment. Migration stimulation by IGFBP-1 was abrogated by pretreatment with a Gly-Arg-Gly-Asp-Ser-Pro (GRGDSP), but not a Gly Arg-Gly-Glu-Ser-Pro (GRGESP) hexapeptide, and by mutation of the RGD domain of IGFBP-1 to Trp-Gly-Asp (WGD). IGFBP-1 treatment caused a rapid localization of immunoreactive FAK to cellular lamellipodia, a rapid increase in phosphorylation of FAK and extracellular-signal regulated kinases 1 and 2. Preincubation of EVT cells with Herbimycin A, a tyrosine kinase inhibitor, abrogated IGFBP-1 effects; whereas an MAPK kinase inhibitor, PD 98059, reduced migration regardless of IGFBP 1 treatment. These results indicate that IGFBP-1 stimulation of EVT cell migration occurs by binding of its RGD domain to the alpha 5 beta 1 integrin, leading to activation of FAK and stimulation of MAPK pathway. PMID- 11397845 TI - Evidence of fetal microchimerism in Hashimoto's thyroiditis. AB - Fetal microchimerism, the engraftment of fetal progenitor cells into maternal tissues, has been implicated in the etiology of autoimmune diseases. We used PCR analysis to determine whether microchimerism occurred in the thyroid glands of female patients suffering from Hashimoto's disease and thus may be involved in its etiology. PCR amplification was performed from thyroid gland specimens using primers unique to a Y-chromosomal sequence (SRY gene) and primers for a sequence that is Y/X-chromosomal homologous except for a 6-bp deletion in the X chromosomal sequence (amelogenin). Microchimerism was detected in 8 of 17 Hashimoto patients, but in only 1 of 25 controls (nodular goiters). Both groups were of similar age and had comparable numbers of pregnancies and numbers of sons. All individuals with microchimerism had given birth to at least 1 son. Our results show that microchimerism is significantly more common in Hashimoto patients than in patients suffering from nodular goiter. We therefore suggest that microchimerism might play a role in the development of Hashimoto's disease, although we cannot completely eliminate the hypothesis that microchimerism is just an "innocent bystander" in a process triggered by other mechanisms. PMID- 11397846 TI - Changes in endothelium-dependent vasodilatation and alpha-adrenergic responses in resistance vessels during the menstrual cycle in healthy women. AB - During the menstrual cycle, changes in endothelium-dependent vasodilatation have been demonstrated in conduit vessels in vivo, but responses in resistance vessels have not been studied. The aim of this study was to examine endothelium-dependent vasodilatation, the effects of local nitric oxide synthesis, and alpha-adrenergic constriction in resistance vessels during the menstrual cycle in 15 healthy female volunteers (mean age, 28.07 +/- 2.1 yr). Forearm blood flow in response to intrabrachial infusion of bradykinin (10, 30, and 100 pmol/min; endothelium dependent vasodilator), glyceryl trinitrate (4, 8, and 16 nmol/min; endothelium independent vasodilator), noradrenaline (60, 120, and 240 pmol/min; alpha adrenergic receptor agonist), and N(G)-monomethyl-L-arginine (1, 2, and 4 micromol/min; nitric oxide synthase inhibitor) was assessed by venous occlusion plethysmography. All subjects were studied in early menstrual phase (days 1--4) and midcycle (days 10-13). Vasodilator response to bradykinin, expressed as the within-subject mean difference in the area under the dose-response curve between phases, was significantly increased at midcycle compared with that in the early menstrual phase (486.5 +/- 165.0; P = 0.01), whereas there was no significant difference in response to glyceryl trinitrate (185.8 +/- 239.0; P = 0.45). The vasoconstrictor response to noradrenaline was significantly greater at midcycle (97.1 +/- 39.4; P = 0.027), but the response to N(G)-monomethyl-L-arginine was not significantly different (17.5 +/- 35.2; P = 0.63). Serum estradiol was approximately 3-fold higher at midcycle, with a mean difference of 252.3 +/- 56.0 pmol/L (P = 0.0005). Progesterone concentrations were not significantly different (-0.11 +/- 0.1 nmol/L; P = 0.28). Differences in endogenous estrogen levels between menstrual phases may underlie changes in bradykinin and noradrenaline responses. If exogenous estrogens have similar effects, the balance of these two opposing actions may determine whether estrogen replacement in postmenopausal women has beneficial or harmful effects on the vasculature. PMID- 11397847 TI - Expression of inflammatory cytokines in placentas from women with preeclampsia. AB - It is postulated that inadequate remodeling of the uterine spiral arteries in preeclampsia leads to focal ischemia and generation of inflammatory cytokines, such as tumor necrosis factor (TNF alpha) and interleukins (ILs), by the placenta. Our objective was to compare TNF alpha, IL-1 alpha, IL-1 beta, and IL-6 levels in placentas from patients with preeclampsia and normal term pregnancies. Because the placenta is a large heterogeneous organ, we analyzed multiple sites per placenta. On the average, there was a 3-fold variation in cytokine protein levels across the eight sites analyzed for each placenta. However, there were no significant overall differences among the normal term, preeclamptic, and preterm placentas from women without preeclampsia. There were also no significant differences in TNF alpha messenger ribonucleic acid between the normal term and preeclamptic placentas, although TNF alpha messenger ribonucleic acid levels were lower in placentas from preterm patients without diagnosis of preeclampsia than in the normal term placentas. In vitro, hypoxia stimulated the production of TNF alpha, IL-1 alpha and IL-1 beta, but not that of IL-6, by placental villous explants from both groups of patients, and this was not exaggerated in preeclampsia. Finally, although peripheral and uterine venous levels of TNF alpha were elevated in preeclamptic women compared with normal term patients, the ratio of uterine to peripheral venous TNF alpha levels was not significantly different from 1.0 for either patient group. Taken together, these results suggest that sources other than the placenta contribute to the elevated concentrations of TNF alpha and IL-6 found in the circulation of preeclamptic women. PMID- 11397848 TI - Adrenocortical secretion of dehydroepiandrosterone in healthy women: highly variable response to adrenocorticotropin. AB - Excess adrenal androgen (AA) levels are observed in 25--50% of women with the polycystic ovary syndrome (PCOS), and AA excess in PCOS may represent selection bias. Thus, it is possible that AA secretion among the general population is highly variable, and that those women who are predisposed to secreting greater amounts of AA have a greater probability of having PCOS. We now hypothesize that the levels of AAs are highly variable among normal nonhyperandrogenic women, and that this heterogeneity is the result of a variable response of AAs to ACTH stimulation. To test this hypothesis we prospectively studied the response of dehydroepiandrosterone (DHA) and cortisol (F) to a 60-min acute stimulation with ACTH-(1--24) in 56 healthy eumenorrheic nonhirsute healthy women with a mean age of 28.9 yr (range, 20--37 yr.) and a mean body mass index (BMI) of 29.2 kg/m(2) (18.2--46.2 kg/m(2)). Baseline samples and poststimulation samples were assayed for DHA and F. The basal and ACTH-stimulated levels of DHA, but not those of F, were negatively correlated with age, although neither the basal nor ACTH stimulated responses of DHA and F varied with BMI. After controlling for age, the basal F level was negatively correlated to its net increment (i.e. Delta F; r = 0.54; P < 0.001), whereas there was no significant relationship between basal DHA and Delta DHA. We also compared the intersubject variability (coefficient of variation) for basal and stimulated levels of DHA and F. For basal (DHA(0)), 60 min (DHA(60)), and net increment in (Delta DHA) DHA levels, the coefficients of variation were 67.9%, 61.4%, and 76.0%, respectively; for F(0), F(60), and Delta F, they were 40.4%, 16.9%, and 31.3%, respectively. The variance in Delta DHA was significantly higher, and the variance in F(60) was significantly lower than that in all other variables; DHA(0), DHA(60), F(0), and Delta F had similar variances. In conclusion, in our population of healthy reproductive-aged women we observed that both basal and ACTH-stimulated levels of DHA after ACTH-(1--24) stimulation had significantly greater intersubject variance (approximately 60--70%) compared with the basal and poststimulation levels of F (approximately 15--40%). These data support the hypothesis that among normal women, AA (i.e. DHA) levels are highly variable compared to those of F. In addition, the intersubject variability in DHA levels is at least in part due to a variable response of AAs to ACTH stimulation. Whether the AA excess frequently observed in PCOS is due to the greater risk of those women with higher AA levels, basally and after ACTH stimulation, remains to be confirmed. PMID- 11397850 TI - Increased vasopressin and adrenocorticotropin responses to stress in the midluteal phase of the menstrual cycle. AB - Accumulating evidence indicates that gonadal steroids modulate functioning of the hypothalamic-pituitary-adrenal (HPA) axis, which has been closely linked to the pathophysiology of anxiety and depression. However, the effect of the natural menstrual cycle on HPA axis responsivity to stress has not been clearly described. In nine healthy women, metabolic and hormonal responses to treadmill exercise stress during the early follicular phase of the menstrual cycle, when gonadal steroid levels are low, were compared with responses in the midluteal phase of the cycle, when both progesterone and estrogen levels are relatively high. Exercise intensity was gradually increased over 20 min to reach 90% of each subject's maximal oxygen consumption during the final 5 min of exercise. Basal plasma lactate, glucose, ACTH, vasopressin, oxytocin, and cortisol levels were similar in the two cycle phases. However, in response to exercise stress, women in the midluteal phase had enhanced ACTH (P < 0.0001), vasopressin (P < 0.01), and glucose (P < 0.001) secretion. These findings suggest that relatively low levels of gonadal steroids during the early follicular phase of the menstrual cycle provide protection from the impact of stress on the HPA axis. PMID- 11397849 TI - Regulation of leptin and leptin receptor in baboon pregnancy: effects of advancing gestation and fetectomy. AB - Leptin, a product of both adipose tissue and the placental syncytiotrophoblast and a potential regulator of primate conceptus development, increases in the maternal circulation with advancing gestation. This increase may be potentiated by estrogens, which also increase as pregnancy progresses. In the present study adipose tissue was collected from nonpregnant (n = 5) baboons (Papio sp) and in baboons during early (days 58-62; n = 5), mid (days 98--102; n = 5), and late (days 158-162; n = 5) pregnancy (term, approximately 184 days). Additionally, placental estrogen production was inhibited in pregnant baboons by the removal of fetal androgen precursors via fetectomy at midgestation, with tissues collected from fetectomized (n = 5) baboons approximately 60 days later. Leptin, estrogens, and androgens were quantitated in maternal serum by RIA. Leptin (LEP) and leptin receptor (LEP-R(L) and LEP-R(S) isoforms) messenger ribonucleic acids (mRNAs) were quantitated by competitive RT-PCR, and leptin concentrations were determined by RIA in maternal adipose and placental villous tissues. Although LEP transcript abundance in adipose tissues was unchanged as a result of pregnancy or with advancing gestation, the leptin protein level was higher (P < 0.02) in pregnant baboons in early gestation than in nonpregnant baboons and increased with gestational age (P < 0.04). Maternal serum estrogens (estradiol and estrone) and androgens (androstenedione and testosterone) were lower (P < 0.0001) in fetectomized baboons than in intact controls. Serum leptin concentrations were unchanged by fetectomy, but the abundance of LEP mRNA transcripts was lower (P < 0.003) in sc adipose tissue and 3-fold higher (P < 0.05) in placenta. Similarly, the leptin protein level declined (P < 0.05) in sc adipose tissue and increased (P < 0.05) in placenta in fetectomized baboons. Although LEP-R(L) mRNA levels were unchanged after fetectomy, placental LEP-R(S) transcript abundance was lower (P < 0.04) than in pregnancy-intact baboons matched for gestational age. Results suggest that both adipose tissue and the placenta may contribute to maternal hyperleptinemia during normal primate pregnancy. Furthermore, the withdrawal of placental steroids results in the enhanced placental leptin production that is commensurate with a decline in production by sc adipose tissue. PMID- 11397851 TI - Differential regulation of inhibin A and inhibin B by luteinizing hormone, follicle-stimulating hormone, and stage of follicle development. AB - Inhibin B and inhibin A exhibit unique patterns of secretion across the follicular phase of the menstrual cycle. To test the hypothesis that the distinct patterns of inhibin B and inhibin A secretion result from differential regulation by LH and FSH, a series of controlled experiments was designed to dissect the specific effects of LH and FSH at distinct stages of follicle development. After GnRH agonist desensitization, women with small antral follicles were treated with recombinant human LH (rhLH), rhFSH, or rhFSH and estradiol (E(2)). rhLH or rhFSH was also administered when follicles reached the preovulatory stage in gonadotropin-stimulated or spontaneous cycles. At the small antral stage of development, rhFSH, but not rhLH, administration increased inhibin B (17.4 +/- 4.6 to 321.0 +/- 97.0 pg/mL; P < 0.05), inhibin A (0.6 +/- 0.1 to 2.6 +/- 0.6 IU/mL; P < 0.05), and E(2) [15.8 +/- 3.6 to 95.3 +/- 26.9 pg/mL (58.0 +/- 13.2 to 349.8 +/- 98.7 pmol/L); P < 0.05]. The inhibin B increase preceded inhibin A by 48 h. Addition of E(2) to FSH resulted in a greater increase in inhibin B (23.2 +/- 6.4 to 865.2 +/- 294.5 pg/mL; P < 0.05) than FSH alone (P < 0.05). At the preovulatory stage, rhLH administration increased inhibin A (15.9 +/- 10.3 to 21.5 +/- 13.7 IU/mL; P < 0.05) and E(2) [669.4 +/- 285.5 to 943.6 +/- 388.1 pg/mL (2457.4 +/- 1048.1 to 3464.0 +/- 1424.7 pmol/L); P < 0.05], but not inhibin B, as did rhFSH administration in spontaneous cycles [E(2): 226.4 +/- 102.7 to 264.7 +/ 121.0 pg/mL (831.1 +/- 377.0 to 971.7 +/- 444.2 pmol/L); P < 0.05; inhibin A: 2.6 +/- 1.3 to 3.7 +/- 1.9 IU/mL; P < 0.05; and inhibin B: 76.3 +/- 32.2 to 77.6 +/- 32.8 pg/mL; P = NS]. These findings suggest that increases in both FSH and E(2) in the early follicular phase result in increased inhibin B secretion at early stages of follicle development, whereas the selective LH rise in the late follicular phase favors inhibin A secretion from more mature follicles. Thus, both differential secretion of LH and FSH and the stage of follicle development determine the patterns of inhibin A and inhibin B secretion in the normal menstrual cycle. PMID- 11397852 TI - Ovarian morphology and serum hormone markers as predictors of ovarian follicle recruitment by gonadotropins for in vitro fertilization. AB - Twenty-five normal ovulatory women underwent three-dimensional transvaginal ultrasonography and blood sampling before and after oral glucose tolerance testing to compare ovarian morphology and circulating hormone levels in the early follicular phase as predictors of the number of oocytes retrieved after gonadotropin stimulation for in vitro fertilization. Serum levels of gonadotropins, inhibins, testosterone, dehydroepiandrosterone sulfate, and estradiol as well as summed ovarian volume were unrelated to oocyte number. Antral follicle number and serum androstenedione level, however, positively correlated, whereas postoral glucose tolerance test (post-OGTT) insulin release negatively correlated, with total and mature oocyte numbers. Adjusting for age and body mass index by regression analysis, the serum androstenedione level significantly predicted mature, but not total, oocyte number. The relationships of antral follicle number and post-OGTT insulin release to total oocyte number were additive; each was significant after controlling for the other. In contrast, antral follicle number significantly correlated with mature oocyte number after controlling for post-OGTT insulin release, whereas post-OGTT insulin release was unrelated to mature oocyte number after controlling for antral follicle number. Therefore, early follicular phase antral follicle number positively correlates with total and mature oocyte numbers after gonadotropin stimulation for in vitro fertilization and is linked to androgen and insulin actions in predicting ovarian follicle recruitment by gonadotropins. PMID- 11397853 TI - Preterm birth: associated neuroendocrine, medical, and behavioral risk factors. AB - Increased CRH secretion by the placenta of pregnant women has been associated with preterm birth. Certain indices of risk, both medical and psychosocial in nature, have been linked to preterm delivery. Levels of total, bound, and free CRH, CRH-binding protein (CRH-BP), and cortisol were measured prospectively in a large sample of pregnant Danish women who delivered preterm and term infants. Measures of maternal serum hormones were taken at 7--23 and 27--37 weeks gestation and, for those who delivered at term, at 37--43 weeks gestation. At 7- 23 weeks gestation, maternal levels of total CRH (P = 0.01), bound CRH (P = 0.03), and CRH-BP (P = 0.01) were higher in the preterm than in the term group. At 27--37 weeks gestation, levels of total CRH (P < 0.0001), bound CRH (P < 0.0001), free CRH (P < 0.0001), and cortisol (P < 0.0001) were all higher in the preterm than the term group, whereas levels of CRH-BP (P < 0.0001) were lower in the preterm than in the term group. The best medical and behavioral factors associated with preterm delivery were, respectively, previous preterm delivery (P < 0.0001) and engagement in certain risk-taking behaviors (P = 0.008). The positive relations between preterm delivery and various adverse medical and socioeconomic variables with increases in placental secretion of CRH suggest that the latter may participate in the pathophysiology of preterm delivery. PMID- 11397854 TI - Interleukin-6 differentially stimulates haptoglobin production by peritoneal and endometriotic cells in vitro: a model for endometrial-peritoneal interaction in endometriosis. AB - Based on our previous observation that peritoneal endometriotic (PE) lesions synthesize in vivo substantially more haptoglobin (Hp) than related eutopic tissues, we hypothesized that this increase in Hp production was due to endometrial-peritoneal interactions. As interleukin-6 (IL-6) stimulates Hp in other tissues and is produced by endometrial cells, we tested our hypothesis by evaluating the effects of IL-6 on Hp production by PE cells, normal peritoneal (P) cells, and eutopic endometrial cells from women with (UE-E) and without endometriosis (UE-C) using semiquantitative RT-PCR and enzyme-linked immunoabsorbent assay. Endogenous production of IL-6 was also assessed. Treatment with human recombinant IL-6 and dexamethasone significantly increased Hp production by P or PE cells in a dose- and time-dependent manner (P < 0.05). Hp messenger ribonucleic acid was not detected in UE-E and UE-C cells in the absence or presence of IL-6 and dexamethasone. PE and UE-E cells expressed significantly more IL-6 messenger ribonucleic acid than P and UE-C cells (P < 0.05). Moreover, UE-E cells secreted 6 times more IL-6 protein than UE-C cells (P < 0.05). These findings support our hypothesis and suggest a novel endometrial-peritoneal interaction whereby locally synthesized IL-6 and Hp may participate in the establishment and persistence of peritoneal endometriosis. PMID- 11397855 TI - Polymorphisms of the androgen receptor gene and the estrogen receptor beta gene are associated with androgen levels in women. AB - To elucidate the possible role of genetic variation in androgen receptor (AR), estrogen receptor alpha (ER alpha), and ER beta on serum androgen levels in premenopausal women, the CAG repeat polymorphism of the AR gene, the TA repeat polymorphism of the ER alpha gene, and the CA repeat polymorphism of the ER beta gene were studied in a population-based cohort of 270 women. Total testosterone, free testosterone, dehydroepiandrosterone sulfate, androstenedione, 17 hydroxyprogesterone, 3 alpha-androstanediol glucuronide, 17 beta-estradiol, LH, FSH, and sex steroid hormone-binding globulin (SHBG) were measured in serum samples obtained in the follicular phase of the menstrual cycle. Women with relatively few CAG repeats in the AR gene, resulting in higher transcriptional activity of the receptor, displayed higher levels of serum androgens, but lower levels of LH, than women with longer CAG repeat sequences. The CA repeat of the ER beta gene also was associated with androgen and SHBG levels; women with relatively short repeat regions hence displayed higher hormone levels and lower SHBG levels than those with many CA repeats. In contrast, the TA repeat of the ER alpha gene was not associated with the levels of any of the hormones measured. Our results suggest that the serum levels of androgens in premenopausal women may be influenced by variants of the AR gene and the ER beta gene, respectively. PMID- 11397856 TI - A unique exonic splicing mutation in the human androgen receptor gene indicates a physiologic relevance of regular androgen receptor transcript variants. AB - In a patient with partial androgen insensitivity syndrome (AIS), we identified a single inherited presumably silent nucleotide variation (AGC -> AGT) in exon 8 (codon 888) of the AR gene. However, in the patient's genital skin fibroblasts, a considerably shortened transcript of 5.5 kb (normal: 10.5 kb) was detected, which misses a part of exon 8 and a prominent portion of the 3'-untranslated region. The translation product includes eight missense amino acids from codon 886 onward followed by a premature stop codon. As shown by in vitro expression analysis, the mutant protein lacks any residual function. However, reverse transcribed PCRs and sequence data indicate the existence of two additional splicing variants of 6.4 kb and 7.8-kb length both in patient and normal control genital skin fibroblasts. These splicing variants comprise the complete coding region but a shortened 3' untranslated region. Thus, a distinct alternative pre-messenger RNA-processing event leading to two additional transcripts occurs generally in genital skin fibroblasts. In addition, this process partially prevents aberrant splicing in the patient and produces a small fraction of normal, functionally intact AR protein that could explain the partial masculinization in this patient. This first report of an exonic splicing mutation in the AR-gene indicates a physiologic relevance of the regular AR-messenger RNA variants with shortened 3' untranslated regions and their functional translation products in human genital development. PMID- 11397857 TI - Inhibin B: comparison with indexes of fertility among formerly cryptorchid and control men. AB - Infertility may be a consequence of cryptorchidism. We previously reported, using a large study cohort, that 38% of formerly bilateral cryptorchid men, 10% of unilateral cryptorchid men, and 5% of the control group were infertile. Men from this cohort donated blood and semen samples for inhibin B, FSH, LH, testosterone, free testosterone, and semen analyses. Results are reported comparing the entire group; some comparisons are based on normal or low sperm density. Data are also presented for men who had fathered children or had unsuccessfully attempted paternity. Mean (+/-SD) inhibin B levels were lower for the cryptorchid men (109 +/- 59 pg/mL) than the control men (153 +/- 60; P < 0.001), and FSH levels were higher (7.4 +/- 6.2 and 4.0 +/- 3.2; P < 0.0001). Inhibin B levels correlated with all other parameters for the cryptorchid group; however, correlations for the control group were only found with gonadotropins. Among the cryptorchid men, levels were significantly greater among men with normal sperm counts than men with low sperm counts (124 +/- 47 vs. 75 +/- 48 pg/mL; P < 0.0001). No difference was present for the control group (155 +/- 61 vs. 149 +/- 63 pg/mL). When the fertile group (based on paternity) vs. the infertile group (based on attempted paternity) were compared, significant differences were found for the cryptorchid group (117 +/- 62 vs. 73 +/- 52 pg/mL; P < 0.03), but not the control group (163 +/- 62 vs. 146 +/- 73 pg/mL). These data reveal relationships not apparent among the control group of men, which includes infertile men. Inhibin B data suggest that a larger portion of formerly cryptorchid men have compromised testicular function than indicated by paternity data. Low levels of inhibin B among individuals are an indication of diminished seminiferous tubule function and thus compromised potential for fertility. Low inhibin B levels together with elevated FSH levels and decreased sperm density are indicative of a high risk of infertility. PMID- 11397858 TI - Inverse correlation between sperm concentration and number of androgen receptor CAG repeats in normal men. AB - Androgens are essential for the maintenance of normal spermatogenesis. Androgen action is mediated by the androgen receptor (AR), which in the testis is expressed by Leydig, peritubular, and Sertoli cells. The fact that sperm numbers range from 20 up to 300 million/mL in normal men without any indication of changed endocrine parameters led us to assume that genetic variability of transduction of androgen signaling could be important. We therefore compared the variable number of CAG repeats in the AR with sperm concentrations in men with normal ejaculate parameters (62 fathers and 69 volunteers participating in clinical trials). In multivariate analysis CAG repeat length did not differ between the volunteers (19.4 +/- 3.1) and the fathers (20.6 +/- 3.0), but was significantly correlated to sperm concentrations with a coefficient of -0.25. When compared with a group of infertile men with (n = 14) or without (n = 30) a family history of infertility, no such correlation was found. These results indicate that men with short CAG repeats have the highest sperm output within the normal fertile population. Polymorphisms of the AR contribute to the efficiency of spermatogenesis in normal men, but do not play a predominant role in male infertility. PMID- 11397859 TI - Metabolic and steroidogenic alterations related to increased frequency of polycystic ovaries in women with a history of gestational diabetes. AB - The prevalence of polycystic ovaries (PCO) and clinical, endocrine, and metabolic features were investigated in women with previous gestational diabetes (GDM). Thirty-three women with a history of GDM and 48 controls were studied. Glucose and insulin secretion capacity was evaluated by means of the oral glucose tolerance test (OGTT), and insulin action was determined by means of a euglycemic insulin clamp. Compared with control women, women with previous GDM more often had significantly abnormal OGTT, a higher prevalence of PCO (39.4% vs. 16.7%; P = 0.03), higher serum concentrations of cortisol, dehydroepiandrosterone, and dehydroepiandrosterone sulfate and a greater area under the glucose curve. Women with previous GDM showed a lowered early phase insulin response to glucose and impaired insulin sensitivity, which was accounted for mainly by decreased glucose nonoxidation. They also demonstrated a significantly lower fasting serum C peptide/insulin ratio than the controls, indicating that women with previous GDM have impaired hepatic insulin extraction, which tended to be more marked among women with PCO. This may explain why women with PCO and previous GDM were significantly more hyperinsulinemic than women with normal ovaries. In conclusion, our data demonstrate that women with previous GDM often have PCO and abnormal OGTT. They are insulin resistant as a result of lowered glucose nonoxidation and show inappropriately low insulin responses to glucose, reflecting impaired beta-cell function. They also have higher adrenal androgen secretion, which may be associated with abdominal obesity. PMID- 11397860 TI - Aromatization mediates testosterone's short-term feedback restraint of 24-hour endogenously driven and acute exogenous gonadotropin-releasing hormone-stimulated luteinizing hormone and follicle-stimulating hormone secretion in young men. AB - The present clinical study examines the neuroregulatory hypothesis that feedback restraint of LH and FSH secretion by testosterone requires in vivo aromatization. To test this postulate, we prospectively and randomly assigned 47 healthy young men to 1 of 5 parallel short-term (5-day) double-blind interventions with: 1) placebo; 2) high-dose ketoconazole (KTCZ, 400 mg orally 4 times daily) to block both Leydig-cell and adrenal steroidogenesis; 3) KTCZ and transdermal testosterone delivery (7.5 mg daily); 4) KTCZ and transdermal estradiol (0.05 mg daily); or 5) KTCZ, testosterone, and the selective and potent aromatase inhibitor, anastrazole (5 mg orally twice daily). Blood was sampled every 10 min for 27 h on the last day of intervention to quantitate 24-h mean spontaneous and 3-h post-GnRH-stimulated (100 ng/kg iv bolus) LH and FSH release. KTCZ administration lowered the serum total testosterone concentration markedly from (mean +/- SEM) 423 +/- 57 ng/dL (15 +/- 2.0 nmo/L) during placebo ingestion to 58 +/- 8.6 ng/dL (2.0 +/- 0.3 nmol/L) (P < 10(-3)). Transdermal androgen addback along with KTCZ blockade increased testosterone levels to 607 +/- 57 ng/dL (21 +/ 2.0 nmol/L). KTCZ exposure alone drove a 3-fold increase in serum LH concentrations (P < 10(-3)) and a 2.5-fold rise in FSH secretion (P = 0.015), as assessed by high-specificity immunoradiometric assays. Concomitant transdermal testosterone (or estradiol) delivery repressed the elevated secretion of both LH and FSH to mid-normal baseline values. A 3-fold administration of anastrazole, KTCZ, and testosterone completely opposed exogenous testosterone's suppression of 24-h LH and FSH secretion. Anastrazole coadministration likewise abolished testosterone-dependent inhibition of 3-h GnRH-stimulated LH and FSH release. In summary, assuming the specificity of anastrazole's inhibition of aromatase activity, we conclude that circulating testosterone in healthy men curtails endogenously driven as well as exogenous GnRH-stimulated LH and FSH secretion conditional on its in vivo aromatization. PMID- 11397861 TI - Human recombinant luteinizing hormone is as effective as, but safer than, urinary human chorionic gonadotropin in inducing final follicular maturation and ovulation in in vitro fertilization procedures: results of a multicenter double blind study. AB - In a prospective, comparative, dose-finding study, the minimal effective dose of recombinant human LH (rhLH) required to induce final follicular maturation and early luteinization in patients undergoing in vitro fertilization and embryo transfer was determined. In addition, the efficacy and safety of rhLH were compared with urinary human CG (u-hCG). A total of 259 infertile women, aged 18 39 yr, were enrolled in the study. After pituitary desensitization using a GnRH agonist, rhFSH was administered for ovarian stimulation. Patients then received either rhLH or u-hCG to achieve final follicular maturation. The doses of rhLH administered were 5,000, 15,000, 30,000, or 15,000 + 10,000 IU (second injection administered 3 days after the first injection; 129 patients), and those of u-hCG were consistently 5,000 IU (121 patients). Ovum pick-up was performed 34--38 h after rhLH or u-hCG injection. After fertilization in vitro, up to three embryos were replaced in the uterine cavity. The numbers of oocytes retrieved after u-hCG or rhLH administration were not significantly different between the four different doses of rhLH, when compared with each corresponding u-hCG group, nor when compared with the pool of all u-hCG groups. Similarly, there were no statistically significant differences in: the number of oocytes retrieved per follicle with a diameter of over 10 mm on the day of u-hCG or rhLH administration; the number of patients with at least one oocyte retrieved; oocyte nuclear maturity; oocyte potential for fertilization; the number of embryos; the number of total, biochemical, and clinical pregnancies; and the embryo implantation rate. However, in many of these parameters, the lowest dose of rhLH seemed suboptimal when compared with the higher dose. In terms of safety, rhLH was well tolerated at a dose of up to 30,000 IU. Moderate ovarian hyperstimulation syndrome (OHSS) was reported in 12.4% of patients who received u hCG and 12.0% of patients who received two injections of rhLH. No moderate or severe OHSS was reported in patients who received a single dose of rhLH up to 30,000 IU. The results show that a single dose of rhLH is effective in inducing final follicular maturation and early luteinization in in vitro fertilization and embryo transfer patients and is comparable with 5,000 IU u-hCG. A single dose of rhLH results in a highly significant reduction in OHSS compared with hCG. The dose of rhLH giving the highest efficacy to safety ratio was between 15,000 and 30,000 IU. PMID- 11397862 TI - Persistence of macroprolactinemia due to antiprolactin autoantibody before, during, and after pregnancy in a woman with systemic lupus erythematosus. AB - A woman with systemic lupus erythematosus (SLE) with marked increases in circulating 150-kDa PRL was studied from before conception, throughout pregnancy, and after pregnancy. The clinical features of the patient included idiopathic hyperprolactinemia without clinical symptoms such as amenorrhea and galactorrhea before pregnancy. No clinical lupus activity was present during follow-up. Serum PRL increase during pregnancy in this patient was considerably higher at weeks 27 and 33 than in normal pregnant women. In contrast, serum-free PRL levels were considerably lower at weeks 20, 27, and 33 than in normal pregnant women. A 150 kDa PRL (big big PRL) species persisted as the predominant circulating form of PRL throughout each measurement in this woman with SLE. In contrast, the predominant form of PRL in serum from healthy pregnant women was little PRL (or monomeric PRL). The nature of big big PRL was due to the presence of anti-PRL autoantibodies forming an IgG-23 kDa PRL complex, in accordance with the studies by affinity chromatography for IgG and Western blot analysis. The IgG-PRL complex was fully bioactive in vitro (Nb2 rat lymphoma cell assay). Injection of the serum into the rats demonstrated that the IgG-PRL complex was cleared more slowly than serum containing predominantly monomeric PRL. The data suggest that the IgG PRL complex has biological activity; the absence of symptoms in this woman may be attributed to the fact that due to its large molecular weight, big big PRL does not easily cross the capillary walls. Delayed clearance may account for increased serum PRL levels in this SLE patient with anti-PRL autoantibodies. PMID- 11397863 TI - Dynamics of ovarian function in an adult woman with McCune--Albright syndrome. PMID- 11397864 TI - Adrenocortical-pituitary hybrid tumor causing Cushing's syndrome. AB - We describe the first case of an adrenocortical-pituitary hybrid tumor causing Cushing's syndrome in a 17-yr-old boy. Adrenal vein sampling confirmed elevated secretion of both cortisol and ACTH precursors from a right adrenal mass, whereas pituitary ACTH levels, as determined by bilateral inferior petrosal sinus samples (IPSS), were unresponsive to CRH and equal to peripheral levels. There was no biochemical or histological evidence for a pheochromocytoma, but, rather, the tumor demonstrated lipid-rich clear cells characteristic of an adrenocortical adenoma. Immunohistochemical analysis revealed ACTH immunoreactivity and synaptophysin proteins in the tumor. Isolation of tumor cells by the novel technique of laser capture microdissection and subsequent RT-PCR showed expression of POMC messenger ribonucleic acid and cytochrome p450 enzyme messenger ribonucleic acid within the same cells. Finally, ultrastructural analysis provided ultimate proof for adrenocortical-pituitary hybrid cells exhibiting the characteristic vesicular mitochondria and abundant smooth endoplasmic reticulum of steroid cells and the typical secretory granules of corticotrophs within the cytoplasm of the same cells. The adrenocortical tumor expressed the pituitary transcription factor pituitary homeobox factor 1 and the steroidogenic factor 1. The intermingling of the centrally located ectodermally derived pituitary tissue with the mesodermally derived adrenocortical tissue in this adenoma suggests a hitherto unrecognized genetic and phenotypic plasticity within the hypothalamic-pituitary-adrenal axis. PMID- 11397865 TI - Double-blind Y chromosome microdeletion analysis in men with known sperm parameters and reproductive hormone profiles: microdeletions are specific for spermatogenic failure. AB - Y chromosome microdeletions have been reported as a possible genetic factor of male infertility. Despite a large number of studies in this subject, there is still considerable debate and confusion surrounding the role of Y chromosome microdeletions in male infertility. This has been further compounded by observations of Y microdeletions in fertile males. The aim of the present study was to evaluate: 1) the incidence of Y microdeletions in control male population and infertile males, where complete semen and hormonal analysis was available to define whether Y microdeletions are specific for spermatogenic failure or if they can be found also in normospermic men; and 2) whether the suboptimal semen quality reported in Denmark is associated with a higher incidence of Y microdeletions in respect to other populations. Double-blind molecular study of deletions was performed in 138 consecutive patients seeking intracytoplasmic sperm injection treatment, 100 men of known fertility, and 107 young military conscripts from the general Danish population. Microdeletions or gene-specific deletions were not detected in normospermic subjects or in subfertile men with a sperm count of more than 1 x 10(6)/mL. Deletions of the Azoospermia factor (AZF)c region were detected in 17% of individuals with idiopathic azoo/cryptozoospermia and in 7% of individuals with nonidiopathic azoo/cryptozoospermia. The data indicate that: 1) the composition of the study population is the major factor in determining deletion frequency; 2) Y chromosome microdeletions are specifically associated with severe spermatogenic failure; therefore, the protocol described here is reliable for the routine clinical workup of severe male factor infertility; and 3) the frequency of Yq microdeletions in the Danish population is similar to that from other countries and argues against the involvement of microdeletions in the relatively low sperm count of the Danish population. PMID- 11397866 TI - The presence of functional luteinizing hormone/chorionic gonadotropin receptors in human sperm. AB - The functional receptors that bind human CG (hCG) and LH have recently been identified in a number of nongonadal human tissues. The current experiments tested the hypothesis that human ejaculated sperm may also contain them. The data revealed that they, indeed, do as determined by the presence of receptor messenger RNA and receptor protein that can bind (125)I-hCG. The receptors were functional, as indicated by an increase in cyclic AMP levels and activation of sperm protein kinase A following treatment with hCG or LH. Treatment with these hormones, on the other hand, had no effect on sperm protein kinase C activity. Now that the functional LH/hCG receptors are found in human sperm, it is important to determine whether hCG treatment could improve the outcome of infertility procedures. PMID- 11397867 TI - Overexpression of type IV collagen in chorionic villi in hydatidiform mole. AB - To investigate the characteristic structure of hydatidiform mole, type IV collagen expression was determined in human villous tissues obtained from normal pregnancies (n = 17) and complete hydatidiform moles (n = 10). Indirect immunofluorescent staining was performed to detect type IV collagen with specific monoclonal antibody, and Northern blot analysis was performed to assess expression of messenger ribonucleic acid for the alpha1(IV) chain. In addition, serum levels of type I, III, and IV collagen were measured by RIA. Immunohistochemical staining for type IV collagen revealed stronger staining of the trophoblastic basement membrane in hydatidiform mole than in normal pregnancy. Northern blot analysis revealed that the villous expression of messenger ribonucleic acid for the alpha1(IV) chain was significantly increased in hydatidiform moles compared with normal pregnancy (P < 0.01). Although there were no differences in the serum type I and III collagen levels between hydatidiform mole and normal pregnancy, the type IV collagen level was significantly higher in patients with hydatidiform mole than in normal pregnancy (P < 0.05). These results suggest that type IV collagen might play an important role in determining the pathophysiology and structure of hydatidiform mole. PMID- 11397868 TI - Hypoxia regulates insulin-like growth factor-binding protein 1 in human fetal hepatocytes in primary culture: suggestive molecular mechanisms for in utero fetal growth restriction caused by uteroplacental insufficiency. AB - Intrauterine growth restriction (IUGR) can be a consequence of decreased uterine blood flow (uteroplacental insufficiency) and maternal and fetal hypoxia. Insulin like growth factors (IGFs) and their binding proteins (IGFBPs) are key elements in fetal growth. IGF-I is a major growth promoter in utero. IGFBP-1 is primarily made in the liver, and it mostly inhibits IGF actions at the cellular level. IGFBP-1 is elevated in the fetal circulation of human and animal pregnancies complicated by IUGR caused by placental insufficiency and in utero hypoxia and is believed to restrict fetal growth by sequestering IGFs. In this study, we developed a protocol to establish highly pure primary cultures of human fetal hepatocytes in vitro and investigated their expression of IGFBP-1 messenger RNA (mRNA) and protein and the effects of hypoxia on their expression of IGFBP-1 mRNA and protein. Hepatocytes were isolated from second-trimester human fetal livers (n = 7) and purified by Percoll gradient centrifugation. Hepatocyte cultures were characterized by immunocytochemistry and were compared with hepatocytes in situ in human fetal liver tissue, by immunohistochemistry, using specific antibodies and indirect immunofluorescence. Cultures consisted primarily (>90%) of cells positive for cytokeratin 18, fibrinogen, and IGFBP-1, with less than 2% vascular cells and less than 8% macrophages. Identification of isolated hepatocytes was further confirmed by morphology. Hepatocytes were cultured in defined medium, and Northern analysis revealed expression of a 1.5-kb IGFBP-1 mRNA transcript in hepatocytes cultured under normoxic conditions, for 24 h, that did not increase in steady-state levels after 48 h in culture. Under hypoxic conditions (2% O(2)), IGFBP-1 mRNA expression increased 3- to 4-fold, compared with normoxic controls. Cells cultured under 10% O(2) did not demonstrate an increase in IGFBP-1 mRNA levels. IGFBP-1 protein in conditioned medium (CM) was measured by immunoradiometric assay and increased 3- to 4-fold under hypoxic (2% O(2)), compared with normoxic, conditions. Western ligand blot analysis of CM revealed the presence of IGFBP-1, IGFBP-2, IGFBP-3, and IGFBP-4. IGFBP-1 was the most abundant IGFBP in CM, and densitometric analysis revealed a 2.5-fold increase in IGFBP-1 under hypoxic, compared with normoxic, conditions, supporting the immunoradiometric assay results. A 3-fold increase in IGFBP-3 mRNA, but not other IGFBPs, was noted under hypoxic, compared with normoxic, conditions. This study demonstrates that human fetal hepatocytes can be cultured in defined medium, as primary cultures with high purity, and that they express IGFBP-1 mRNA and secrete IGFBP-1 protein in vitro. In addition, the data demonstrate that hypoxia up regulates fetal hepatocyte IGFBP-1 mRNA steady-state levels and protein, with this being the major IGFBP derived from the fetal hepatocyte. The data support a role for the fetal liver as a source of elevated circulating levels of IGFBP-1 in fetuses with in utero hypoxia and IUGR. PMID- 11397869 TI - Interleukin-8 in the human fallopian tube. AB - The human fallopian tube is a dynamic structure that undergoes cyclic variation in its functional epithelium. This epithelium contains both secretory and ciliated cells. The mechanisms regulating the growth and function of the tubal epithelium are not fully understood. Interleukin-8 (IL-8) is one potential local regulatory factor. We therefore characterized the IL-8 system, which includes IL 8, its receptors A and B, and its degradative enzyme aminopeptidase N, in the human fallopian tube by immunohistochemistry. Immunohistochemistry was performed on isthmic, ampullary, and fimbrial fallopian tubal segments obtained from women undergoing gynecological surgical procedures for benign conditions (n = 52). IL-8 was found in the human fallopian tube predominantly in the epithelial cells. It was present in greater amounts in the distal compared with the proximal tube. IL 8 receptors A and B localized in the tube in similar patterns. The degradative enzyme aminopeptidase N is found in tubal stromal tissue at the epithelial stromal border and perivascularly and may limit the systemic effects of epithelial IL-8. The IL-8 system seems to be an active component of tubal physiology. PMID- 11397870 TI - Progesterone antagonists increase androgen receptor expression in the rhesus macaque and human endometrium. AB - Antiprogestins (APs) inhibit estradiol (E(2))-stimulated endometrial growth in women and nonhuman primates, but the mechanism of this "antiestrogenic" action is unknown. Here, we report that APs up-regulate endometrial androgen receptor (AR) in both women and macaques, an effect that might play a role in the antiproliferative effects of APs on the primate endometrium. In addition, because there are discrepancies in the literature on the regulation and localization of AR in the primate endometrium, we used both in situ hybridization and immunocytochemistry to evaluate hormonal influences on endometrial AR in women and macaques. In ovariectomized macaques, the following treatments were given for 4 weeks each: E(2) alone, E(2) + progesterone (P), E(2) + mifepristone (RU 486), and E(2) + P + RU 486. In women, samples were obtained during the normal menstrual cycle and after treatment with either RU 486 for 30 days at 2 mg/day, or after a single oral administration of 200 mg RU 486 on cycle day LH + 2. In macaques, E(2) significantly increased AR expression above vehicle controls; E(2) + RU 486 increased binding further; E(2) + P decreased AR binding; and E(2) + P + RU 486 treatment caused an intermediate elevation in AR binding. In macaques treated with E(2) alone, stromal AR staining was predominant, and P treatment suppressed that staining. E(2) + RU 486 or E(2) + P + RU 486 treatment produced a striking up-regulation of glandular epithelial AR staining and enhanced the stromal AR signal. In situ hybridization analyses confirmed the immunocytochemistry data. Similar induction of glandular AR staining and enhanced stromal AR staining were obtained in macaques treated with ZK 137 316 and ZK 230 211. During the natural cycle in women, stromal AR staining predominated and was greater in the proliferative than the late secretory phase. RU 486 treatment of women up-regulated glandular epithelial AR staining after either daily treatment for 30 days with 2 mg/day or after a single oral dose of 200 mg. In summary, endometrial AR was highest in the stroma during the human proliferative phase (or during E(2) treatment in macaques) and lowest during the late secretory phase in women (or after E(2) + P treatment in macaques). In both species, RU 486 induced AR expression in the glands and enhanced AR expression in stromal cells. Because androgens can antagonize E(2) action, enhanced endometrial AR expression induced by APs could play a role in the antiproliferative, "antiestrogenic" effects of APs in primates. PMID- 11397871 TI - Two novel mutations in the gonadotropin-releasing hormone receptor gene in Brazilian patients with hypogonadotropic hypogonadism and normal olfaction. AB - Several point mutations in the GnRH receptor gene have been described in an autosomal recessive form of congenital isolated hypogonadotropic hypogonadism (HH). We investigated 17 Brazilian patients (10 males and 7 females) from 14 different families, with HH and normal olfaction. The diagnosis of HH was based on absent or incomplete sexual development after 17 yr of age associated with low or normal levels of LH in both sexes and low levels of testosterone in males and of estradiol in females. All patients presented with a normal sense of smell in an olfactory specific test. The coding region of the GnRH receptor gene was amplified by PCR and directly sequenced. A novel missense mutation, Arg(139)His, located in the conserved DRS motif at the junction of the third transmembrane and the second intracellular loop of the GnRH receptor was identified in the homozygous state in one female with complete HH. The Arg(139)His mutation completely eliminated detectable GnRH-binding activity and prevented GnRH-induced stimulation of inositol phosphate accumulation in vitro. In another family, a new compound heterozygous mutation (Asn(10)Lys and Gln(106)Arg) was identified in four siblings (two males and two females) with partial HH. The Gln(106)Arg mutation, located in the first extracellular loop, has been previously described, and in vitro analysis indicated that the mutant receptor was able to bind GnRH, but with a reduced affinity. The Asn(10)Lys mutation in the extracellular amino terminal domain of the receptor also reduced the affinity for GnRH in vitro. In this family we also identified a previously described silent polymorphism at amino acid residue 151 in the second intracellular loop that segregated with the two inactivating mutations of the GnRH receptor. This polymorphism was also found in two unrelated patients with sporadic HH without GnRH receptor loss of function mutations. No mutations were identified in the remaining cases. A good correlation between genotype and phenotype was found in our patients. The woman, who is homozygous for the completely inactivating Arg(139)His mutation, has complete HH with undetectable serum basal LH and FSH levels that failed to respond to GnRH stimulation. In addition, the affected patients who are compound heterozygotes for the Asn(10)Lys/Gln(106)Arg mutations, have partial HH with low serum basal LH levels that were responsive to GnRH stimulation. No clinical or hormonal differences were found between HH patients with and without mutations in the GnRH receptor gene, indicating that these data do not contribute to the identification of HH patients with GnRH receptor mutations. In conclusion, we report the first naturally occurring mutation within the conserved DRS motif of the GnRH receptor in a female with complete HH and a novel compound heterozygous mutation (Asn(10)Lys and Gln(106)Arg) in a family with partial HH, increasing the repertoire of the inactivating mutations of the GnRH receptor. PMID- 11397872 TI - Normal human pituitary gland and pituitary adenomas express cannabinoid receptor type 1 and synthesize endogenous cannabinoids: first evidence for a direct role of cannabinoids on hormone modulation at the human pituitary level. AB - Little is known about the expression and function of cannabinoid receptor type 1 (CB1) in the human pituitary gland. The aim of this study was to investigate CB1 expression in human normal and tumoral pituitaries by in situ hybridization and immunohistochemistry using an antibody against CB1. CB1 was found in corticotrophs, mammotrophs, somatotrophs, and folliculostellate cells in the anterior lobe of normal pituitary. After examination of 42 pituitary adenomas, CB1 was detected in acromegaly-associated pituitary adenomas, Cushing's adenomas, and prolactinomas, whereas faint or no expression was found in nonfunctioning pituitary adenomas. Experiments with cultured pituitary adenoma cells showed that the CB1 agonist WIN 55,212--2 inhibited GH secretion in most of acromegaly associated pituitary adenomas tested and that the CB1 antagonist SR 141716A was generally able to reverse this effect. Moreover, WIN 55,212--2 was able to suppress GHRH-stimulated GH release, and this effect was not blocked by coincubation with SR 141716A, possibly indicating a non-CB1-mediated effect. In contrast, WIN 55,212--2 was ineffective on GH-releasing peptide-stimulated GH release. In four Cushing's adenomas tested, WIN 55,212--2 was not able to modify basal ACTH secretion. However, simultaneous application of CRF and WIN 55,212--2 resulted in a synergistic effect on ACTH secretion, and this effect could be abolished by SR 141716A, demonstrating a CB1-mediated effect. In the single case of prolactinomas tested, WIN 55,212--2 was able to inhibit basal secretion of PRL. Finally, the presence of endocannabinoids (anandamide and 2 arachidonoylglycerol) was investigated in normal and tumoral pituitaries. All tumoral samples had higher contents of anandamide and 2-arachidonoylglycerol compared with the normal hypophysis. Moreover, endocannabinoid content in the different pituitary adenomas correlated with the presence of CB1, being elevated in the tumoral samples positive for CB1 and lower in the samples in which no or low levels of CB1 were found. The results of this study point to a direct role of cannabinoids in the regulation of human pituitary hormone secretion. PMID- 11397873 TI - Rapid communication: predominant intracellular overexpression of the Na(+)/I(-) symporter (NIS) in a large sampling of thyroid cancer cases. AB - Here we report the analysis of the Na(+)/I(-) symporter (NIS) protein expression in 57 thyroid cancer samples by immunohistochemistry with high-affinity anti-NIS Abs. As many as 70% of these samples exhibited increased NIS expression with respect to the normal surrounding thyroid tissue. Most significantly, NIS was located in these samples either in both the plasma membrane and intracellular compartments simultaneously, or exclusively in intracellular compartments. This suggests that NIS is clearly expressed or even overexpressed in most thyroid cancer cells, but malignant transformation in some of these cells interferes either with the proper targeting of NIS to the plasma membrane, or with the mechanisms that retain NIS in the plasma membrane after it has been targeted. The results further indicate that, in addition to indicating NIS expression in cases where it is absent (approximately 30%), improvements in (131)I radioablation therapy might result from promoting targeting of NIS to the plasma membrane in the majority (approximately 70%) of thyroid cancers. PMID- 11397874 TI - Congenital adrenal hyperplasia due to 21-hydroxylase deficiency: alterations in cortisol pharmacokinetics at puberty. AB - In congenital adrenal hyperplasia (CAH) due to 21-hydroxylase deficiency, treatment with glucocorticoid and mineralocorticoid substitution is not always satisfactory. Suboptimal control is often observed in pubertal patients, despite adequate replacement doses and adherence to treatment. We investigated whether the pubertal process is associated with alterations in cortisol pharmacokinetics resulting in a loss of control of the hypothalamic-pituitary-adrenal axis. We determined the pharmacokinetics of hydrocortisone administered iv as a bolus. A dose of 15 mg/m(2) body surface area was given to 14 prepubertal (median age, 9.4 yr; range, 6.1--10.8 yr), 20 pubertal (median, 13.5 yr; range, 10.6--16.8 yr), and 6 postpubertal (median, 18.2 yr; range, 17.2--20.3 yr) patients with salt wasting CAH. All patients were on standard replacement therapy with hydrocortisone and 9 alpha-fludrocortisone. Serum total cortisol concentrations were measured at 10-min intervals for 6 h following iv hydrocortisone bolus and analyzed using a solid-phase RIA. The serum total cortisol clearance curve was monoexponential. Mean clearance was significantly higher in the pubertal group (mean, 427.0 mL/min; SD, 133.4) compared with the prepubertal (mean, 248.7 mL/min; SD, 100.6) and postpubertal (mean, 292.4 mL/min; SD, 106.3) (one-way ANOVA, F = 9.8, P < 0.001) groups. This effect persisted after adjustment for body mass index. The mean volume of distribution was also significantly higher in the pubertal (mean, 49.5 L; SD, 12.2) than the prepubertal (mean, 27.1 L; SD, 8.4) patients but not in the postpubertal (mean, 40.8 L; SD, 16) (ANOVA, F = 15.2, P < 0.001) patients. The significance remained after correction for body mass index. There was no significant difference in mean half-life of total cortisol in prepubertal (mean, 80.2 min; SD, 19.4), pubertal (mean, 84.4 min; SD, 24.9), and postpubertal (mean, 96.7 min; SD, 9.9) patients. Similar differences between groups were observed when the pharmacokinetic parameters of free cortisol were examined. In addition, the half-life of free cortisol was significantly shorter in females compared with males (P = 0.04). These data suggest that puberty is associated with alterations in cortisol pharmacokinetics resulting in increased clearance and volume of distribution with no change in half-life. These alterations probably reflect changes in the endocrine milieu at puberty and may have implications for therapy of CAH and other conditions requiring cortisol substitution in the adolescent years. PMID- 11397875 TI - Tie-2 is expressed on thyroid follicular cells, is increased in goiter, and is regulated by thyrotropin through cyclic adenosine 3',5'-monophosphate. AB - Angiogenesis is coordinated with follicular cell growth in goitrogenesis. The angiopoietins, Ang-1 and Ang-2, are angiogenic growth factors acting through Tie 2, a tyrosine kinase receptor. We have examined the expression and regulation of the angiopoietins and Tie-2 in human and rat thyroids. In human goiters there was increased Tie-2 immunostaining, compared with that in normal thyroids, on both follicular and endothelial cells. In an induced goiter in rats, in situ hybridization showed increased expression of messenger ribonucleic acids (mRNAs) for Tie-2 and Ang-1 in follicular cells. As Tie-2 has previously been believed to be restricted to cells of endothelial lineage in adults, we examined its expression further in isolated follicular cells. Tie-2 and Ang-1 mRNA expression in human thyrocytes was confirmed by ribonuclease protection assay. Ang-2 mRNA was not detected in human cultures or rat thyroids. In both human follicular cell cultures and FRTL-5 cells, immunoblotting showed that Tie-2 expression was increased by TSH and agents that increased intracellular cAMP. In conclusion, we have demonstrated the expression of Tie-2 and Ang-1 in thyroid epithelial and endothelial cells, and have shown the regulation of Tie-2 by TSH and cAMP in follicular cells. Tie-2 expression is increased in goiter in both humans and rats, consistent with a role in goitrogenesis. PMID- 11397876 TI - Adipose tissue metabolism in benign symmetric lipomatosis. AB - Type 2 benign symmetric lipomatosis (BSL) is characterized by abnormal growth of adipose tissue in the upper back, deltoid region, upper arms, hips, and upper thigh region. Studies of lipomatous tissue in vitro have suggested that defective lipolysis may account for excess fat accumulation; however, in vivo adipose tissue metabolism has not been evaluated. We measured systemic adipose tissue lipolysis and regional adipose tissue fatty acid uptake in a patient with type 2 BSL scheduled for elective brachioplasty. We found increased, rather than decreased, rates of systemic free fatty acid release coupled with normal fatty acid oxidation. The uptake of fatty acids was 19% greater in deltoid region lipomatous tissue than in abdominal sc fat, whereas in control studies the relative uptake of fatty acids in deltoid fat averaged 29% less than that in abdominal fat. Adipocyte size was smaller than expected in lipomatous tissue. These results suggest that type 2 BSL is a hyperplastic adipose tissue abnormality that does not impair systemic lipolysis. The pathophysiology appears similar to what has been termed hyperplastic obesity. A better understanding of this condition could lead to insights into the mechanisms of hyperplastic obesity. PMID- 11397877 TI - Retinoid receptors in the human endometrium and its disorders: a possible modulator of 17 beta-hydroxysteroid dehydrogenase. AB - Retinoids have recently been proposed to modulate estrogenic actions in various sex steroid-dependent neoplasms, but little has been studied in human endometrial disorders. Therefore, in this study, we first examined the immunolocalization of retinoic acid receptor alpha, beta, and gamma, and retinoid X receptor (RXR) alpha, beta, and gamma in 20 normal cycling human endometria, 34 endometrial hyperplasia, and 46 endometrioid endometrial adenocarcinomas. We then correlated these findings with other clinicopathological parameters, especially in the correlation between retinoid receptor subtypes and the status of steroid hormone receptors, 17 beta-hydroxysteroid dehydrogenase (17 beta-HSD) and aromatase. We also then examined the effects of retinoic acid on the expression of 17 beta-HSD type 2 in cell lines derived from endometrial carcinoma using Northern blotting analysis to examine the possible roles of retinoids in in situ endometrial estrogen metabolism. Among these six retinoid receptors examined, RXR gamma immunoreactivity was exclusively detected in the epithelial cells of the secretory phase endometrium but not of the proliferative phase, which was well correlated with 17 beta-HSD type 2 immunolocalization. However, in endometrial hyperplasia, RXR gamma was not correlated with 17 beta-HSD type 2. In endometrioid endometrial adenocarcinoma, there was a statistically significant correlation between 17 beta-HSD type 2 immunoreactivity and RXR gamma labeling index (LI) (P < 0.001) and between RXR gamma LI and progesterone receptor LI (r = 0.501, P = 0.003). A significant inverse correlation was also detected between RXR gamma LI and patient age (r = 0.449, P = 0.015). No statistically significant correlation was obtained between LIs of receptors and other clinicopathological parameters including the status of intratumoral aromatase examined by immunohistochemistry. In the endometrial carcinoma cell line, RL95-2, retinoic acid markedly increased the level of 17 beta-HSD type 2 messenger RNA in a time- and dose-dependent manner. These results all suggest that retinoic acids may be involved in modulation of in situ estrogen metabolism in both normal and neoplastic human endometrium possibly through RXR gamma by stimulating the expression of 17 beta-HSD type 2. PMID- 11397878 TI - Expression of 11 beta-hydroxysteroid dehydrogenase isoenzymes in the human pituitary: induction of the type 2 enzyme in corticotropinomas and other pituitary tumors. AB - One of the defining biochemical features of Cushing's disease is a relative insensitivity to glucocorticoid (GC) feedback, but an analysis of the GC receptor has failed to detect any major abnormalities. However, two isoenzymes of 11 beta hydroxysteroid dehydrogenase (11 beta HSD), either by converting cortisone (E) to cortisol (F) (type 1) or conversely by converting F to E (type 2), play an important prereceptor role in regulating corticosteroid hormone action at several sites. 11 beta HSD1 and -2 expression within the anterior pituitary gland itself may modulate GC feedback at an autocrine level, and we have speculated that this may be deranged in Cushing's disease. Detection of 11 beta HSD type 1 and 2 immunoreactive protein was performed using fluorescence immunohistochemistry. Double immunofluorescent studies were undertaken on normal pituitary to define the cellular localization of 11 beta HSD isoenzymes using antisera against GH, ACTH, LH, FSH, PRL, and S100, a nonhormonal marker of folliculo-stellate cells. In normal pituitary, positive staining for 11 beta HSD1-immunoreactive protein was observed in GH- and PRL-secreting cells and in folliculo-stellate cells; gonadotrophs, thyrotrophs, and ACTH-positive cells were negative. 11 beta HSD2 immunoreactivity was absent in all cell types. RT-PCR detected 11 beta HSD1 messenger ribonucleic acid (mRNA) expression in the normal pituitary; 11 beta HSD2 mRNA expression was also seen in most normal tissue. By contrast, in ACTH secreting adenomas 11 beta HSD2 immunostaining was strongly positive in every case of corticotroph adenoma. 11 beta HSD1 immunoreactivity was also observed occasionally, but to a much lesser extent. In other pituitary tumors, both functional and nonfunctional, 11 beta HSD expression was variable in terms of isoenzyme mRNA and intensity of protein staining. The expression of 11 beta HSD1 (which generates F from E) in somatotrophs and lactotrophs suggests an autocrine role for this isoenzyme in the glucocorticoid regulation of pituitary GH and PRL secretion. 11 beta HSD2 expression is markedly induced in ACTH-secreting pituitary tumors and, by converting F to E, may explain the resetting of glucocorticoid feedback control in Cushing's disease. PMID- 11397879 TI - Sulfation of thyroid hormone and dopamine during human development: ontogeny of phenol sulfotransferases and arylsulfatase in liver, lung, and brain. AB - Sulfation is an important mechanism for regulating the biological activity of numerous hormones and neurotransmitters in man. Here we have investigated the ontogeny of sulfotransferases (SULT) and sulfatase (ARS) involved in the metabolism of thyroid hormone and dopamine. SULT1A1 enzyme activity was lower in postnatal liver and lung than in fetal tissues. Hepatic SULT1A3 (dopamine) was expressed at high levels early in development, but decreased substantially in late fetal/early neonatal liver and was essentially absent from the adult liver. In lung, significant SULT1A3 activity was observed in the fetus, but neonatal levels were considerably lower. In brain, the highest activity was observed in the choroid plexus for SULT1A1, with low and widespread activity for both SULT1A1 and SULT1A3 in other brain regions. SULT activity with 3,3'-diiodothyronine (3,3' T(2)) as substrate was measured in all tissues and correlated significantly with SULT1A1 activity (4-nitrophenol), suggesting that SULT1A1 is primarily responsible for the sulfation of this iodothyronine. The developmental expression of SULT1A3 and SULT1A1 in liver and brain was confirmed by immunoblot, and immunohistochemistry of developing liver showed substantial expression of these proteins in hemopoietic cells in fetal liver. We also detected low activity for the hydrolysis of 3,3'-T(2) sulfate by ARS, although there was less distinction between fetal and neonatal samples than with SULT activities. We have therefore shown that the developing fetus has substantial sulfation capacity. Sulfation may therefore play a major role in the homeostasis of hormones and other endogenous compounds as well as in detoxification in the fetus, particularly as other conjugating enzyme systems, such as the UDP-glucuronosyltransferases, are not expressed at significant levels until the neonatal period. PMID- 11397880 TI - Laboratory and clinical experience in 55 patients with macroprolactinemia identified by a simple polyethylene glycol precipitation method. AB - PRL exists in different forms in human serum. The predominant form is little PRL (molecular mass 23 kDa) with smaller amounts of big PRL (molecular mass 50--60 kDa) and at times big big or macroprolactin (molecular mass 150--170 kDa). The frequency and clinical consequences of macroprolactinemia have not been clearly established, mainly because of difficulty in identifying these patients biochemically. This previously required the use of gel filtration chromatography, which could not be used routinely. Recently, a screening test using polyethylene glycol (PEG) has been used to identify macroprolactin in serum. Consequently, this study was designed to examine the use of PEG precipitation in the identification of patients with a predominance of macroprolactin and to establish the clinical characteristics of such a cohort. Over 12 months, 18,258 requests for serum PRL were received and of these 1225 patients had a serum PRL more than 700 mU/L. A total of 322 of these patients (26%) had a percentage recovery after PEG precipitation of less than 40%, thus indicating the presence of a predominance of macroprolactin. Fifty-five of these patients were referred for detailed clinical assessment. Symptoms typical of hyperprolactinemia were not common in this cohort. None had sustained amenorrhea and eight have had oligomenorrhea at age less than 40 yr. One had galactorrhea. All had pituitary imaging, and four had a microadenoma with none having a macroadenoma. PEG precipitation allows easy identification of macroprolactin in routine clinical practice. As the clinical consequences of this entity at this stage seem relatively benign, referral and intensive investigation of these patients may not be necessary. However, follow-up of a large cohort is required to ensure that the long-term outlook is likewise benign. This would have important implications for both patients and healthcare systems. PMID- 11397881 TI - Common genomic variation in LMNA modulates indexes of obesity in Inuit. AB - We discovered that rare mutations in LMNA, which encodes lamins A and C, underlie autosomal dominant Dunnigan-type familial partial lipodystrophy. Because familial partial lipodystrophy is an extreme example of genetically disturbed adipocyte differentiation, it is possible that common variation in LMNA is associated with obesity-related phenotypes. We subsequently discovered a common single nucleotide polymorphism (SNP) in LMNA, namely 1908C/T, which was associated with obesity related traits in Canadian Oji-Cree. We now report association of this LMNA SNP with anthropometric indexes in 186 nondiabetic Canadian Inuit. We found that physical indexes of obesity, such as body mass index, waist circumference, waist to hip circumference ratio, subscapular skinfold thickness, and subscapular to triceps skinfold thickness ratio were each significantly higher among Inuit subjects with the LMNA 1908T allele than in subjects with the 1908C/1908C genotype. For each significantly associated obesity-related trait, the LMNA 1908C/T SNP genotype accounted for between approximately 10--100% of the attributable variation. The results indicate that common genetic variation in LMNA is an important determinant of obesity-related quantitative traits. PMID- 11397882 TI - Prevalence of neuroendocrine dysfunction in patients recovering from traumatic brain injury. AB - Although hypopituitarism is a known complication of head injury, it may be underrecognized due to its subtle clinical manifestations. The nonspecific symptoms may be masked by and may contribute to the physical and psychological sequelae of brain trauma. This study examines the prevalence of neuroendocrine abnormalities in patients rehabilitating from traumatic brain injury. Seventy adults (mean age, 31.5 +/- 1.1 yr; range, 18--58; 46 men and 24 women) with traumatic brain injury an average of 49 +/- 8 months before the study (median, 13 months) underwent a series of standard endocrine tests, including serum levels of TSH, free T(4), insulin-like growth factor I, PRL, testosterone (males), and cosyntropin stimulation. Abnormal results of these tests were followed by dynamic tests of gonadotropin, TSH, and GH secretion. Glucagon stimulation testing in 48 subjects revealed GH deficiency (peak, <3 microg/L) in 14.6%. Free T(4) (n = 6; 8.6%), TSH (n = 7; 10%), or both (n = 2; 2.9%) were low in 21.7%, whereas 87% had both TSH and free T(4) below the midnormal level. Basal morning cortisol was below normal in 45.7% of subjects, whereas cosyntropin-stimulated levels were insufficient (peak, <500 nmol/L) in 7.1%. Hypogonadism and hyperprolactinemia were uncommon. In summary, pituitary hormone deficiencies were identified in a substantial proportion of patients with previous brain injury. GH deficiency, found in 15% by glucagon stimulation testing, may compound the physical and psychological complications of traumatic brain injury and interfere with rehabilitation. PMID- 11397883 TI - Serum lipid profile improved by ultra-low doses of 17 beta-estradiol in elderly women. AB - To determine whether ultra-low doses of estradiol (E(2)) affect the serum lipid profile in elderly women, we analyzed changes in serum lipids and lipoproteins in 70 healthy women, 60 yr and older, randomly assigned to parenteral E(2) (7.5 microg per 24 h) delivered by a vaginal ring (Estring; Pharmacia-Upjohn, Malmo, Sweden) or no treatment for 12 months. Baseline serum estrone sulfate (E1S), but not E(2) or serum FSH, was negatively associated with serum total cholesterol (P = 0.026), low-density lipoprotein (LDL) cholesterol (P = 0.053), and apolipoprotein B levels (P = 0.023). Compared with no treatment, Estring treatment yielded nonsignificant increases within the normal postmenopausal range in serum E1S (+16%) and E(2) (+13%), but significantly reduced serum LDL cholesterol by 7.6% (-0.32 mmol/L; 95% confidence interval, -0.58, -0.07; P = 0.014) and LDL to high-density lipoprotein (HDL) ratio by 7.3% (-0.19 mmol/L; 95% confidence interval, -0.44, -0.06; P = 0.030). In Estring users values were significantly reduced in total cholesterol (by 4%), LDL cholesterol (by 7%), LDL to HDL ratio (by 7%), and apolipoprotein B (by 4%), and significantly increased in serum HDL triglyceride (by 25%) but not triglycerides. No significant changes were found in the untreated group. There was a significant interaction between age and both baseline serum E(2)/sex hormone-binding globulin (P = 0.006) and sex hormone-binding globulin (P = 0.009) and a marginal interaction between age and E1S (P = 0.083) with regard to effects on changes in LDL cholesterol levels during Estring treatment. We conclude that ultra-low doses of E(2), which previously were considered to have only local effects, may improve serum lipid profile in elderly women with a pattern and magnitude similar to that reported after conventional estrogen doses or first-generation lipid-lowering agents. The reduction in LDL cholesterol tended to be greater with a combination of high age and low baseline levels of biologically active estrogens. PMID- 11397884 TI - Effects of the selective estrogen receptor modulator, raloxifene, on the somatotropic axis and insulin-glucose homeostasis. AB - Raloxifene is the first selective estrogen receptor modulator registered for the prevention and treatment of postmenopausal osteoporosis. In addition to direct effects on bone cells, estrogen and raloxifene may act indirectly via changes in hormonal homeostasis. However, the menopause-related decrease in serum insulin like growth factor I (IGF-I) and the increase in insulin or glucose are not always reversed by estrogen replacement. Especially orally administered estrogen was reported to decrease serum IGF-I levels. Understanding the effects of estrogens and raloxifene on the GH-IGF axis and insulin-glucose homeostasis are important because of their link to bone metabolism and cardiovascular health. We investigated the effects of raloxifene on the GH-IGF-I axis and insulin-glucose homeostasis in a cross-sectional study in the third year of the Multiple Outcomes of Raloxifene Evaluation trial, a double blind, placebo-controlled, prospective study in postmenopausal women with osteoporosis (T-score of -2.5 or less or at least two moderate vertebral fractures). Patients with diabetes mellitus were excluded from this additional study. A fasting blood sample was obtained (0 h), and women received an sc injection of 0.05 mg recombinant human GH (Humatrope)/kg BW. The second blood sample was obtained 24 h later (24 h). GH, IGF-I, IGF binding protein-3 (IGFBP-3), insulin, and glucose were measured. Group characteristics were tested by nonparametric ANOVA. The dose-response to raloxifene was tested by linear regression models, with age and body mass as covariates. Seven women were taking placebo, 16 were taking raloxifene (60 mg/day), and 9 were taking raloxifene (120 mg/day). Patients from the 60 mg raloxifene group were the oldest (mean +/- SD, 64.4 +/- 4.2 vs. 69.3 +/- 6.9 and 63.3 +/- 5.9 yr for placebo, 60 mg/day raloxifene, and 120 mg/day raloxifene, respectively; P = 0.05). Compared with placebo users, patients taking raloxifene had higher body mass index (24.7 +/- 1.7 vs. 25.0 +/- 3.1 and 28.8 +/- 5.8 kg/m(2); P = 0.03). At 0 h, raloxifene use was associated with lower IGF-I/IGFBP 3 ratio (4.3 +/- 0.7 vs. 2.9 +/- 0.7 and 3.0 +/- 0.7 nmol/mg; P = 0.001) and insulin/glucose ratio (13.7 +/- 5.2 vs. 11.9 +/- 5.9 and 9.5 +/- 2.3 pmol/mmol; P = 0.04). Similarly, raloxifene use was associated with lower IGF-I/IGFBP-3 and insulin/glucose ratios at 24 h (P = 0.01 and 0.07). Glucose, GH, and IGFBP-3 levels were similar among the groups (0.12 < P < 0.67). In conclusion, raloxifene use is associated with decreased serum IGF levels and insulin/glucose ratio before and 24 h after one rhGH injection in nondiabetic postmenopausal women with osteoporosis. Therefore, raloxifene may decrease liver sensitivity to GH. Other explanations are increased clearance or increased tissue sensitivity to IGF-I or insulin. The raloxifene-induced increases in bone mineral density do not appear to be mediated by reversing the age- and menopause-related decreases in IGF-I levels. The results of this small cross-sectional study need confirmation by longitudinal studies. PMID- 11397885 TI - Role of free fatty acids on cardiac autonomic nervous system in noninsulin dependent diabetic patients: effects of metabolic control. AB - Decreased heart rate variability (HRV) is a risk factor for cardiovascular mortality. Elevated plasma free fatty acid (FFA) levels decrease HRV in healthy subjects. Thus, we investigated the effect of changes in plasma FFA levels on HRV, in non-insulin-dependent diabetes (NIDDM) patients. Thirty NIDDM patients free from diabetic neuropathy volunteered for a study made by two phases. In study A, changes in HRV along a 10% lipid emulsion infusion + heparin (n = 15) or saline infusion (control study; n = 15) were investigated. In study B, all patients (n = 30) underwent further determination of HRV after 3 months of improved metabolic control achieved by intensified insulin treatment. In study A, lipid emulsion infusion increased plasma FFA (P < 0.001) and catecholamine concentrations (P < 0.005), mean arterial blood pressure (P < 0.005), low frequency/high frequency (LF/HF) ratio (P < 0.001). Delta plasma FFA levels correlated with delta LF/HF ratio (r = 0.57; P < 0.02). Along with saline infusion, metabolic and cardiovascular parameters remained unchanged throughout the test. In study B, improved metabolic control lowered fasting plasma glucose (P < 0.005), FFA (P < 0.001), norepinephrine (P < 0.02), epinephrine (P < 0.04), and glycosylated hemoglobin levels (P < 0.001), mean arterial blood pressure(P < 0.05), and LF/HF ratio (P < 0.001). Again percent decline in plasma FFA correlated with the percent change in LF/HF ratio (r = 0.72; P < 0.001). In a multivariate analysis, percent changes in LF/HF ratio were associated with percent changes in plasma FFA independently of gender and percent changes in body mass index, waist/hip ratio, plasma norepinephrine, epinephrine, glycosylated hemoglobin, and daily insulin therapy. Our study demonstrates that changes in plasma FFA levels may have a parallel effect on cardiac sympathetic/parasympathetic nervous system balance in NIDDM patients. PMID- 11397886 TI - Premature adiposity rebound in children treated for acute lymphoblastic leukemia. AB - The adiposity rebound (AR), when body mass index begins to increase after its nadir in childhood, is a critical period for the regulation of energy balance and adult obesity risk. The aim of the present study was to test whether children treated for acute lymphoblastic leukemia (ALL) experience premature AR. This might, in part, explain their tendency to develop obesity. Timing of AR was assessed by visual inspection of body mass index plots in 68 patients treated for ALL in first remission. This sample comprised all eligible patients treated in Scotland between 1991 and 1998, age 30 months or less at the time of diagnosis. Timing of AR in patients was compared against a cohort of 889 healthy British children studied during the 1990s using the same method. AR occurred significantly earlier in the patients treated for ALL (chi(2) test, P < 0.001). The AR had occurred in 43% (29 of 68) of the patients and 4% (40 of 889) of the comparison group by age 37 months. At 49 months AR had occurred in 81% (55 of 68) of the patients and 21% (190 of 889) of the comparison group. Treatment of ALL is associated with a significantly advanced AR. This might, in part, explain the extremely high prevalence of obesity in long-term survivors. Clinical management should focus on minimizing excess weight gain during therapy to reduce long-term obesity risk. PMID- 11397887 TI - Long-term effects of depot long-acting somatostatin analog octreotide on hormone levels and tumor mass in acromegaly. AB - The effects of a 12- to 24-month treatment with depot long-acting octreotide (OCT LAR) on hormone profile, tumor mass, and clinical symptoms were reported in 36 patients with active acromegaly [GH, 34.2 +/- 5.6 microg/L; insulin-like growth factor I (IGF-I), 784.5 +/- 40.4 microg/L]. Fifteen patients were de novo whereas 21 had previously undergone unsuccessful surgery. Serum GH (P < 0.0001) and IGF-I levels (P < 0.0001) significantly decreased as early as after the first injection of OCT-LAR and progressively declined during the 12-24 months of treatment both in de novo and in operated patients. At the last follow-up, GH hypersecretion was controlled (< or =2.5 microg/L) in 69.4% whereas normal IGF-I levels were achieved in 61.1% of patients. GH and IGF-I suppression during OCT-LAR treatment was similar in de novo and operated patients as shown by nadir GH (2.3 +/- 0.6 vs. 2.2 +/- 0.6 microg/L) and IGF-I (323.1 +/- 34.9 vs. 275.5 +/- 33.0 microg/L), percent suppression of GH (92.7 +/- 2.0 vs. 85.9 +/- 3.3%) and IGF-I (57.4 +/- 4.9 vs. 61.5 +/- 4.6%), and prevalence of GH (73.3 vs. 76.2%) and IGF-I (53.3 vs. 71.4%) control. A decrease in tumor volume was observed in 12 of 15 de novo patients, whereas no shrinkage was detected in 4 of 9 operated patients. No patient had tumor reexpansion during OCT-LAR treatment. Significant clinical improvement was obtained in all patients; heart rate, systolic blood pressure, and diastolic blood pressure significantly decreased in the entire population. A mild but significant increase of blood glucose levels, followed by a decrease of serum insulin levels, was observed after 3 months of treatment: this effect subsided with treatment continuation. OCT-LAR treatment was well tolerated by most patients. In conclusion, long-term treatment with OCT-LAR was effective in controlling GH and IGF-I hypersecretion in most patients with acromegaly, when applied either as primary therapy or as adjunctive therapy after surgery. Tumor shrinkage was observed in de novo patients during OCT-LAR treatment, suggesting that it can be successfully applied as primary therapy in patients bearing invasive tumors, who are less likely to be cured after surgery. PMID- 11397888 TI - Bone loss in men with prostate cancer treated with gonadotropin-releasing hormone agonists. AB - Prostate cancer is the most common visceral malignancy in men. As the tumor is testosterone dependent, a frequent treatment modality involves therapy with GnRH agonists (GnRH-a) resulting in hypogonadism. Because testosterone is essential for the maintenance of bone mass in men, we postulated that GnRH-a therapy would negatively impact skeletal integrity. We compared bone mineral density (BMD), biochemical markers of bone turnover, and body composition in 60 men with prostate cancer (19 men receiving GnRH-a therapy and 41 eugonadal men) and BMD in 197 community-living healthy controls of similar age. BMD was assessed by dual energy x-ray absorptiometry and ultrasound. Biochemical markers of bone turnover, included markers of bone resorption (urinary N-telopeptide) and bone formation markers (bone-specific alkaline phosphatase and osteocalcin). Body composition (total body fat and lean body mass) was assessed by dual energy x-ray absorptiometry. Significantly lower BMD was found at the lateral spine (0.69 +/- 0.17 vs. 0.83 +/- 0.20 g/cm(2); P < 0.01), total hip (0.94 +/- 0.14 vs. 1.05 +/- 0.16 g/cm(2); P < 0.05), and forearm (0.67 +/- 0.11 vs. 0.78 +/- 0.07 g/cm(2); P < 0.01) in men receiving GnRH-a compared with the eugonadal men with prostate cancer. Significant differences were also seen at the total body, finger, and calcaneus (all P < 0.01). BMD values in eugonadal men with prostate cancer and healthy controls were similar. Markers of bone resorption (urinary N-telopeptide) and bone formation (bone-specific alkaline phosphatase) were elevated in men receiving GnRH-a therapy compared with those in eugonadal men with prostate cancer. Men receiving GnRH-a also had a higher percent total body fat (29 +/- 5% vs. 25 +/- 5%; P < 0.01) and lower percent lean body weight (71 +/- 5% vs. 75 +/- 5%; P < 0.01) compared with eugonadal men with prostate cancer. In conclusion, men with prostate cancer receiving androgen deprivation therapy have a significant decrease in bone mass and increase in bone turnover, thus placing them at increased risk of fracture. PMID- 11397889 TI - Linkage of the human inducible nitric oxide synthase gene to type 1 diabetes. AB - Exposure of human pancreatic islets to a mixture of cytokines induces expression of the inducible nitric oxide synthase (iNOS), impairs beta-cell function, and induces apoptosis. We performed a mutational scanning of all 27 exons of the human NOS2 gene and linkage transmission disequilibrium testing of identified NOS2 polymorphisms in a Danish nationwide type 1 diabetes mellitus (IDDM) family collection. Mutational screening was performed using PCR-amplified exons, followed by single stranded conformation polymorphism and verification of potential polymorphisms by sequencing. The transmission disequilibrium test was performed in an IDDM family material comprising 257 Danish families; 154 families were affected sibling pair families, and 103 families were simplex families. In total, 10 polymorphisms were identified in 8 exons, of which 4 were tested in the family material. A C/T single nucleotide polymorphism in exon 16 resulting in an amino acid substitution, Ser(608)Leu, showed linkage to IDDM in human leukocyte antigen DR3/4-positive affected offspring (P = 0.008; corrected P = 0.024). No other distorted transmission patterns were found for any other tested single nucleotide polymorphism or constructed haplotypes with the exception of those including data from exon 16. In conclusion, linkage of the human NOS2 gene to IDDM in a subset of patients supports a pathogenic role of nitric oxide in human IDDM. PMID- 11397890 TI - Fitness, training, and the growth hormone-->insulin-like growth factor I axis in prepubertal girls. AB - We recently demonstrated that a brief endurance type training program led to increases in thigh muscle mass and peak oxygen uptake (VO(2)) in prepubertal girls. In this study, we examined the effect of training on the GH-->insulin-like growth factor I (GH-->IGF-I) axis, a system known to be involved both in the process of growth and development and in the response to exercise. Healthy girls (mean age 9.17 +/- 0.10 yr old) volunteered for the study and were randomized to control (n = 20) and training groups (n = 19) for 5 weeks. Peak VO(2), thigh muscle volume, and blood samples [for IGF-I, IGF-binding proteins (IGFBP)-1 to 6, and GHBP] were measured. At baseline, IGF-I was significantly correlated with both peak VO(2) (r = 0.44, P < 0.02) and muscle volume (r = 0.58, P < 0.004). IGFBP-1 was negatively correlated with muscle volume (r = -0.71, P < 0.0001), as was IGFBP-2. IGFBP-4 and -5 were significantly correlated with muscle volume. We found a threshold value of body mass index percentile (by age) of about 71, above which systematic changes in GHBP, IGFBP-1, and peak VO(2) per kilogram were noted, suggesting decreases in the following: 1) GH function, 2) insulin sensitivity, and 3) fitness. Following the training intervention, IGF-I increased in control (19.4 +/- 9.6%, P < 0.05) but not trained subjects, and both IGFBP-3 and GHBP decreased in the training group (-4.2 +/- 3.1% and -9.9 +/- 3.8%, respectively, P < 0.05). Fitness in prepubertal girls is associated with an activated GH-->IGF-I axis, but, paradoxically, early in a training program, children first pass through what appears to be a neuroendocrine state more consistent with catabolism. PMID- 11397891 TI - Interaction between beta-adrenergic receptor stimulation and nitric oxide release on tissue perfusion and metabolism. AB - Nitric oxide (NO) may be an important modulator of sympathetic tone. We used im and sc microdialysis in humans to characterize the interaction of NO synthase inhibition and adrenoreceptor stimulation on tissue perfusion, metabolism, and norepinephrine release. Microdialysis probes were perfused with L- or D-nitro-L arginine-methyl-ester (100 micromol/L) followed by incremental doses of isoproterenol, epinephrine, or nitroprusside. Blood flow was estimated based on the ethanol dilution technique. In muscle, the increase in blood flow with isoproterenol was abolished by L-NAME. The ethanol ratio was 0.03 +/- 0.011 with D-NAME and 0.075 +/- 0.014 with L-NAME during isoproterenol treatment (1 micromol/L). The effect was less pronounced in adipose tissue. The vasodilatory effect of nitroprusside was similar with D- and L-NAME. L-NAME augmented isoproterenol- and epinephrine-induced glycerol release. Dialysate glycerol during 1 micromol/L isoproterenol was 47 +/- 6.7 micromol/L with D-NAME and 72 +/ 15 micromol/L with L-NAME. In skeletal muscle, dialysate norepinephrine during 1 micromol/L isoproterenol treatment was 0.73 +/- 0.17 and 1.3 +/- 0.15 nmol/L with D- and L-NAME, respectively. We conclude that NO synthase inhibition attenuates beta(2)-adrenoreceptor-mediated vasodilation and enhances beta-adrenoreceptor mediated lipolysis. These effects are in part mediated through an increase in interstitial norepinephrine concentrations. The data are consistent with the idea that in humans, NO is important in modulating and ameliorating sympathetic effects in peripheral tissues. PMID- 11397892 TI - Macrophage migration inhibitory factor and hypothalamo-pituitary-adrenal function during critical illness. AB - In patients with septic shock (n = 32), multitrauma (n = 8), and hospitalized matched controls (n = 41), we serially measured serum macrophage inhibitory factor (MIF), cortisol, plasma ACTH, tumor necrosis factor-alpha, and interleukin 6 (IL-6) immunoreactivity during 14 days or until discharge/death. MIF levels were significantly elevated on day 1 in septic shock (14.3 +/- 4.5 microg/L), as opposed to trauma (3.1 +/- 1.7 microg/L) and control patients (2.5 +/- 2.1 microg/L). The time course of MIF, parallel to cortisol, but in contrast to ACTH, showed persistently elevated levels in septic patients. On admission, nonsurvivors of septic shock (n = 11) showed significantly higher MIF levels than survivors (18.4 +/- 4.8 and 10.2 +/- 4.2 microg/L, respectively). Patients with septic adult respiratory distress syndrome (ARDS; n = 8) showed higher MIF levels than those who did not develop ARDS (19.4 +/- 4.7 vs. 9.2 +/- 4.3 microg/L, respectively). Multiple logistic regression analysis demonstrated that both MIF and ARDS were independent predictors of adverse outcome. On admission, tumor necrosis factor-alpha, IL-6, procalcitonin, and lipopolysaccharide-binding protein levels were higher in patients with septic shock than in patients with multitrauma. In septic patients, regression analysis showed significant correlations between MIF and cortisol as well as between MIF and IL-6 levels and disease severity scores. No relation was found between MIF and markers of the acute phase response (procalcitonin, C- reactive protein, and lipopolysaccharide binding protein). In multitrauma patients, MIF levels were not elevated at any time point and were not related to other variables. Our data suggest that during immune-mediated inflammation (such as septic shock) MIF is an important neuroendocrine mediator: a contraregulator of the immunosuppressive effects of glucocorticoids. PMID- 11397893 TI - Dexamethasone inhibits tumor necrosis factor-alpha-induced apoptosis and interleukin-1 beta release in human subcutaneous adipocytes and preadipocytes. AB - Tumor necrosis factor-alpha (TNF alpha) can decrease adipose tissue mass, but in obesity, adipose tissue hypertrophy persists despite increased TNF alpha expression. The hormonal milieu of obesity may antagonize the adipostat effects of TNF alpha. We examined the effects of insulin and the synthetic glucocorticoid, dexamethasone (Dex), on TNF alpha-induced apoptosis and gene expression in human adipocytes and preadipocytes. Using RT multiplex PCR, the expression of the proapoptotic genes interleukin-1 beta (IL-1 beta)-converting enzyme (ICE) and TNF alpha and the antiapoptotic genes bcl-2, nuclear factor kappa B (NF kappa B), and NF kappa B inhibitory subunit, I kappa B, were examined. The expression and release of IL-1 beta, a postulated downstream effector of ICE-mediated apoptosis, were also determined. TNF alpha increased the messenger ribonucleic acid levels of ICE, TNF alpha, IL-1 beta, bcl-2, and NF kappa B in preadipocytes and adipocytes (P < 0.01). Dex inhibited TNFalpha induced messenger ribonucleic acid expression of ICE, TNF alpha, and IL-1 beta (P < 0.01), but not that of bcl-2 and NF kappa B. TNF alpha stimulated IL-1 beta release from preadipocytes and adipocytes up to 20-fold, but the effect was abrogated by Dex. Apoptosis induced by TNF alpha was reduced to control levels (P < 0.01) by Dex. Insulin had no significant effect on TNF alpha-induced apoptosis and gene expression. In obesity, glucocorticoids may reduce TNF alpha actions in adipose tissue by inhibiting TNF alpha-induced apoptosis, IL-1 beta release, and TNF alpha expression. PMID- 11397894 TI - Prolactin (PRL)-releasing peptide stimulates PRL secretion from human fetal pituitary cultures and growth hormone release from cultured pituitary adenomas. AB - The hypothalamic peptide PRL-releasing peptide (PrRP) has recently been cloned and identified as a ligand of an orphan pituitary receptor that stimulates in vitro PRL secretion. PrRP also induces PRL release in rats in vivo, especially in normal cycling females. However, no information on the effects of PrRP in the human is available. To elucidate the role of PrRP in regulating human anterior pituitary hormones, we used human PrRP-31 in primary cultures of human pituitary tissues, including fetal (20--27 weeks gestation) and normal adult pituitaries, as well as PRL- and GH-secreting adenomas. PrRP increased PRL secretion from human fetal pituitary cultures in a dose-dependent manner by up to 35% (maximal effect achieved with 10 nM), whereas TRH was slightly more potent for PRL release. Coincubation with estradiol resulted in enhanced fetal PRL response to PrRP, and GH release was only increased in the presence of estradiol. Although PRL secretion from PRL-cell adenomas was not affected by PrRP, PrRP induced PRL release from cultures of a GH-cell adenoma that cosecreted PRL. PrRP enhanced GH release in several GH-secreting adenomas studied by 25--27%, including GH stimulation in a mixed PRL-GH-cell tumor. These results show for the first time direct in vitro effects of PrRP-31 on human pituitary cells. PrRP is less potent than TRH in releasing PRL from human fetal lactotrophs and is unable to release PRL from PRL-cell adenomas in culture, but stimulated GH from several somatotroph adenomas. Thus, PrRP may participate in regulating GH, in addition to PRL, in the human pituitary. PMID- 11397895 TI - The contribution of intraabdominal fat to gender differences in hepatic lipase activity and low/high density lipoprotein heterogeneity. AB - Hepatic lipase (HL) hydrolyzes triglyceride and phospholipid in low and high density lipoprotein cholesterol (LDL-C and HDL-C, respectively), and elevated HL activity is associated with small, dense atherogenic LDL particles and reduced HDL2-C. Elevated HL activity is associated with increasing age, male gender, high amounts of intraabdominal fat (IAF), and the HL gene (LIPC) promoter polymorphism (C nucleotide at -514). We investigated the mechanisms underlying the difference in HL activity between men (n = 44) and premenopausal women (n = 63). Men had significantly more IAF (144.5 +/- 80.9 vs. 66.5 +/- 43.2 cm(2), respectively; P < 0.001), higher HL activity (220.9 +/- 94.7 vs.129.9 +/- 53.5 nmol/mL.min; P < 0.001), more dense LDL (Rf, 0.277 +/- 0.032 vs. 0.300 +/- 0.024; P = 0.01), and less HDL2-C (0.19 +/- 0.10 vs. 0.32 +/- 0.16 mmol/L; P < 0.001) than women. After adjusting for IAF and the LIPC polymorphism, men continued to have higher (but attenuated) HL activity (194.5 +/- 80.4 vs.151.0 +/- 45.2, respectively; P = 0.007) and lower HDL2-C (0.23 +/- 0.11 vs. 0.29 +/- 0.14 mmol/L; P = 0.02) than women. Using multiple regression, HL activity remained independently related to IAF (P < 0.001), gender (P < 0.001), and the LIPC genotype (P < 0.001), with these factors accounting for 50% of the variance in HL activity. These data suggest that IAF is a major component of the gender difference in HL activity, but other gender-related differences, perhaps sex steroid hormones, also contribute to the higher HL activity seen in men compared with premenopausal women. The higher HL activity in men affects both LDL and HDL heterogeneity and may contribute to the gender difference in cardiovascular risk. PMID- 11397896 TI - The POU homeodomain protein Oct-1 binds Cis-regulatory element essential for the human GnRH upstream promoter activity in JEG-3 cells. AB - In previous studies, we have localized four specific nuclear protein-binding elements in the human GnRH upstream promoter. To test whether these four elements are reproductive tissue specific, we placed the four elements upstream to a thymidine kinase (TK) promoter/luciferase reporter gene, and transfected the constructs into human placental choriocarcinoma (JEG-3) cells. The 272-bp fragment (-994 to -723) containing the four elements can drive heterologous TK promoter expression in JEG-3 cells about 15 times more than that of basal TK promoter activity. Deletion of element 4 (E4, -987/-968) significantly decreased (4-fold) the luciferase activity. Further deletion of the other elements (E3 individual, -960/-940 or E3 and E2 in combination, -919/-896) only slightly decreased the luciferase activity. In contrast, deletion of element 1 (E1, -876/ 851) caused a 2-fold loss of luciferase activity and elimination of E2 and E3 only lost less than 2-fold of the luciferase activity. Study performed with 5' end deletion of this region confirmed these observations. Furthermore, E4 DNA protein complex can be supershifted by Oct-1 antibody, indicating that Oct-1 binds to E4. These results clearly demonstrated that all four elements are required to confer tissue-specific expression of the hGnRH gene in JEG-3 cells. However, the E4 is the most important for the tissue-specific expression of the hGnRH gene in JEG-3 cells. Oct-1 factor binds with E4 element and may be involved in the mediation of the human GnRH upstream promoter activity. PMID- 11397897 TI - The spectrum of molecular defects of the CYP21 gene in the Hellenic population: variable concordance between genotype and phenotype in the different forms of congenital adrenal hyperplasia. AB - Defective steroid synthesis due to 21-hydroxylase deficiency is the most common form of congenital adrenal hyperplasia. Knowledge of the molecular defects causing 21-hydroxylase deficiency in different populations is of both theoretical and practical interest. The types and the relative frequencies of molecular defects and the correlation between the genotype and the phenotype were examined in the Hellenic population. We searched for deletions, conversions, and 11 of the most frequent mutations of the CYP21 gene by Southern blot and allele-specific PCR in 222 chromosomes from 111 unrelated subjects and their parents. The most frequent molecular defects were 1) in the salt wasting form, I(2) splice (42.9%), deletions and conversions (24.5%), and Q318stop (14.3%); 2) in the simple virilizing form, I172N (35.3%), I(2) splice (29.4%), and P30L (19.1%); and 3) in the nonclassical form, V281L (41.1%), P30L (21.4%), and P453S (14.3%). Compared with other populations, Greek patients had a higher frequency of Q318stop in the salt-wasting form, of P30L in both simple virilizing and nonclassical forms and of P453S in the nonclassical form. The concordance of genotype to phenotype in the total sample was 87%. However, the concordance rate was different in the three forms of the disease. Thus, complete concordance was detected in the genotypes predicting the salt-wasting phenotype, a slightly lower concordance (95.2%) was detected in the genotypes predicting the simple virilizing phenotype, and the lowest concordance (67.6%) was observed in genotypes predicting the nonclassical phenotype. In conclusion, the concordance between genotype and phenotype decreases as the severity of the disease diminishes. This should be taken into consideration in genetic counseling and antenatal intervention. PMID- 11397898 TI - Efficacy of the long-acting octreotide formulation (octreotide-LAR) in patients with thyrotropin-secreting pituitary adenomas. AB - The presence of somatostatin receptors on TSH-secreting pituitary adenomas allows treatment of central hyperthyroidism with somatostatin analogs. Six women and 5 men (mean +/- SEM age, 43 +/- 3 yr) presented TSH-secreting pituitary adenomas (micro, n = 2; macro, n = 9). Seven patients had previously been treated with partial surgical removal (n = 6) and/or external radiation (n = 4) of their adenoma at least 1 yr before the study, whereas 4 patients had not been treated before somatostatin analog therapy. TSH, free T(4), and free T(3) levels were in the normal range during treatment with sc injections (n = 9) or continuous infusion (n = 2) of octreotide (280 +/- 25 microg/day). Mean thyroid hormone levels increased (P < 0.01) after the washout period (34 +/- 6 days). The patients received monthly im injections of 20 mg Octreotide-LAR. In patients with an elevated free T(4) level after 3 months (n = 1) the Octreotide-LAR dose was increased to 30 mg. After 3 months of Octreotide-LAR treatment, TSH, free T(4)/T(3), and alpha-subunit levels decreased, and 10 patients were euthyroid with normal free T(4) levels. These results remained at the same level over the next 3 months. There were no statistically significant differences in the TSH and free T(4) responses to sc octreotide or im Octreotide-LAR between previously untreated patients and patients who had undergone surgical resection and/or pituitary radiation before somatostatin analog treatment. During Octreotide-LAR treatment, minor digestive problems or moderate discomfort at the injection site, lasting less than 48 h, were reported in 6 and 5 patients, respectively. Gallbladder echographies did not reveal new gallstones during Octreotide-LAR treatment. In conclusion, this study shows that monthly im Octreotide-LAR is as effective as daily sc octreotide in controlling hyperthyroidism in patients with TSH-secreting pituitary adenomas, in both previously untreated patients and patients treated with surgery and/or pituitary radiotherapy. Octreotide-LAR is well tolerated, except for minor digestive problems or mild pain at the injection site. Therefore, Octreotide-LAR appears to be a useful therapeutic tool to facilitate medical treatment of TSH-secreting pituitary adenomas in patients who need long-term somatostatin analog therapy. PMID- 11397899 TI - Children with organic growth hormone deficiency have elevated cortisol responses to stimuli. AB - The aim of this study was to investigate hypothalamic-pituitary-adrenal (HPA) function in children with GH deficiency. Ninety-four patients were evaluated for GH deficiency and cortisol (F) deficiency using clinical criteria and L-dopa and insulin-induced hypoglycemia stimulation tests. They were assigned to three diagnostic groups: organic GH deficient (OGHD), idiopathic GH deficient (IGHD), and not GH-deficient (NGHD). Time series, cross-sectional, regression analysis revealed statistically significantly elevated F [>828 nmol/L (30 microg/dL)] in the OGHD group vs. the NGHD group. The value for F in the IGHD group was not different from the NGHD group. This finding suggests that dysregulation of the HPA axis is present in most children with OGH deficiency and significantly less often in children with IGH deficiency or without GH deficiency. Anatomical disruption of the control pathways for the HPA axis or stress may cause the dysregulation. PMID- 11397901 TI - Comparison of simple measures of insulin sensitivity in young girls with premature adrenarche: the fasting glucose to insulin ratio may be a simple and useful measure. AB - Insulin resistance is a strong predictor of the development of type 2 diabetes mellitus and cardiovascular disease. Girls with premature adrenarche (PA) or obesity may be at an increased risk for the development of insulin resistance. Recently, in prepubertal girls with PA, a fasting glucose to insulin ratio (FGIR) of less than 7 was found to be predictive of insulin resistance as determined by the frequently sampled iv glucose tolerance test. We sought to compare the FGIR with 2 insulin sensitivity measures, SiM (an adjusted mean measure of insulin sensitivity based on fasting and 2 h post glucose load insulin sensitivity measures) and the composite whole body insulin sensitivity index, ISI(comp), both derived from the 2-h oral glucose tolerance test in 2 groups of children at risk: girls with PA and obese girls. We studied 25 prepubertal girls with PA and/or obesity and further classified them as insulin resistant (IR) or insulin sensitive (IS) based on the FGIR. Four simple measures of insulin sensitivity [FGIR, quantitative insulin sensitivity check index (QUICKI), fasting insulin resistance index, and fasting insulin] were compared with SiM and ISI(comp). Additionally, we characterized the subjects in terms of risk factors associated with insulin resistance according to their insulin resistance status based on the FGIR. In our subjects the strongest correlations overall appeared to be between FGIR and SiM, FGIR and ISI(comp), QUICKI and SiM, and QUICKI and ISI(comp) [correlations (r) ranged from 0.81--0.84]. Furthermore, the IR group had higher body mass index and body mass index z-scores and triglyceride levels than the IS group and were over 3 times more likely to have triglycerides greater than the 95th percentile compared with national norms. We conclude that the FGIR and QUICKI are highly correlated with oral glucose tolerance test measures of insulin sensitivity. An FGIR less than 7 in young girls with PA or obesity may be helpful in the early identification of children at risk for complications of insulin resistance. PMID- 11397900 TI - Potassium depletion and salt sensitivity in essential hypertension. AB - To evaluate the actual role of potassium depletion on blood pressure, 11 hypertensive patients were placed on a 10-day isocaloric diet providing a daily potassium intake of either 18 or 80 mmol, with each subject serving as his or her own control; the intake of sodium (220 mmol/day) and other minerals was kept constant. On day 11 each patient was also subjected to central volume expansion by water immersion associated with either normal or low potassium intake. After a 10-day period of low potassium intake, systolic blood pressure increased (P < 0.02) by 5 mm Hg, whereas serum potassium decreased (P < 0.001) by 0.9 mmol/L; no significant changes in urinary sodium and a marked increase in urinary calcium excretion (P < 0.001) were found during the 10-day low potassium intake. PRA (P < 0.02) and plasma aldosterone (P < 0.04) concentrations also decreased during low potassium intake in hypertensive patients. Even though an identical natriuretic response was found during the water immersion experiments with either high or low potassium in the whole hypertensive group, the evaluation of hypertensive subjects in relation to salt sensitivity enabled us to disclose pronounced differences in the natriuretic and calciuretic response. In fact, although an impaired natriuretic ability and moderate calcium loss were particularly found during water immersion in those hypertensive subjects exhibiting a lower salt sensitivity index, a predominant calcium depletion appeared to be the most important consequence of potassium depletion in the hypertensive subjects with a higher salt sensitivity index. By confirming that potassium depletion may exacerbate essential hypertension, our data also suggest that not only sodium restriction, but also potassium and calcium supplementation, could be particularly advisable in salt-sensitive hypertensive patients. PMID- 11397902 TI - The effect of aromatase inhibition on sex steroids, gonadotropins, and markers of bone turnover in older men. AB - There is evidence that estrogen decreases bone turnover in men as well as women. We therefore hypothesized that older men would show increased bone resorption in response to inhibition of the aromatase enzyme, which converts androgens to estrogen. Fifteen eugonadal men over 65 yr were treated for 9 weeks with 2.0 mg/day of anastrozole, an aromatase inhibitor. After 9 weeks of treatment, there were significant decreases in estradiol, estrone, and sex hormone-binding globulin levels by 29%, 73%, and 16%, respectively, and total testosterone increased significantly by 56%. Despite the limited decrease of estrogen and the increase in testosterone, C-telopeptide of type 1 collagen showed a progressive significant increase of 11%, 24%, and 33% (P for trend = 0.033) above baseline at 3, 6, and 9 weeks, respectively. N-telopeptide of type 1 collagen values were highly correlated with C-telopeptide of type 1 collagen, but the change in N telopeptide of type 1 collagen was not statistically significant. Bone-specific alkaline phosphatase and N-terminal type I procollagen peptides showed significant decreases of 8% and 11% of baseline at 9 weeks. Osteocalcin decreased significantly by 30% at 18 weeks. We conclude that aromatase inhibition can reduce estrogen levels in older men, but this effect is limited, perhaps because of feedback stimulation of testosterone production, and that endogenous estrogen derived from aromatization of testosterone plays a role in bone metabolism of older men by limiting the rate of bone resorption. PMID- 11397903 TI - 5 alpha-reductase type 2 is constitutively expressed in the dermal papilla and connective tissue sheath of the hair follicle in vivo but not during culture in vitro. AB - Recent studies suggest that 5 alpha-reductase type 2 (5 alpha R2) rather than 5 alpha R1 plays a key role in the pathogenesis of male-pattern baldness. To clarify the localization of the androgen receptor (AR), 5 alpha R1, and 5 alpha R2 in the hair follicle, we investigated the expression of the corresponding genes by RT-PCR using microdissected hair follicles. AR and 5 alpha R1 mRNAs were expressed in all portions of the hair follicle. By contrast, 5 alpha R2 mRNA was expressed only in mesenchymal portions that included the dermal papilla and connective tissue sheath, and hardly any was expressed in epithelial portions. The intensity of expression of these genes in each portion of the hair follicles did not differ between follicles from balding and nonbalding scalp. We also examined the expression of these genes in cultured fibroblasts derived from the dermal papilla and connective tissue sheath. Although expression of AR and 5 alpha R1 mRNAs was easily detected, there was no obvious expression of 5 alpha R2 mRNA in either type of cell. Type-specific inhibition of 5 alpha R activity by MK386 and MK906 confirmed these patterns of expression of 5 alpha R mRNA. Thus, the expression of 5 alpha R2 mRNA seems to be characteristic of freshly microdissected mesenchymal portions of the hair follicle, but such expression might not be maintained in culture. PMID- 11397904 TI - Cortisol and growth hormone responses to exercise at different times of day. AB - Exercise of appropriate intensity is a potent stimulus for GH and cortisol secretion. Circadian and diurnal rhythms may modulate the GH and cortisol responses to exercise, but nutrition, sleep, prior exercise patterns, and body composition are potentially confounding factors. To determine the influence of the time of day on the GH and cortisol response to acute exercise, we studied 10 moderately trained young men (24.1 +/- 1.1 yr old; maximal oxygen consumption, 47.9 +/- 1.4 mL/kg.min; percent body fat, 13.2 +/- 0.6%). After a supervised night of sleep and a standard meal 12 h before exercise, subjects exercised at a constant velocity (to elicit an initial blood lactate concentration of approximately 2.5 mmol/L) on a treadmill for 30 min on 3 separate occasions, starting at 0700, 1900, and 2400 h. Blood samples were obtained at 5-min intervals for 1 h before and 5 h after the start of exercise; subjects were not allowed to sleep during this period. Subjects were also studied on 3 control days under identical conditions without exercise. There were no significant differences with time of day in the mean blood lactate and submaximal oxygen consumption values during exercise. The differences over time in serum GH and cortisol concentrations between the exercise day and the control day were determined with 95% confidence limits for each time of day. Exercise stimulated a significant increase in serum GH concentrations over control day values for approximately 105--145 min (P < 0.05) with no significant difference in the magnitude of this response by time of day. The increase in serum GH concentrations with exercise was followed by a transient suppression of GH release (for approximately 55--90 min; P < 0.05) after exercise at 0700 and 1900 h, but not at 2400 h. Although the duration of the increase in serum cortisol concentrations after exercise was similar (approximately 150--155 min; P < 0.05) at 0700, 1900, and 2400 h, the magnitude of this increase over control day levels was greatest at 2400 h. This difference was significant for approximately 130 min and approximately 40 min compared to exercise at 1900 and 0700 h, respectively (P < 0.05). The cortisol response to exercise at 0700 h was significantly greater than that at 1900 h for about 55 min (P < 0.05). A rebound suppression of cortisol release for about 50 min (P < 0.05) was observed after exercise at 2400 h, but not 0700 or 1900 h. Both baseline (before exercise) and peak cortisol concentrations were significantly higher at 0700 h than at 1900 or 2400 h (P < 0.01). We conclude that time of day does not alter the GH response to exercise; however, the exercise-induced cortisol response is modulated by time of day. PMID- 11397905 TI - Analysis of the SDHD gene, the susceptibility gene for familial paraganglioma syndrome (PGL1), in pheochromocytomas. AB - Pheochromocytomas are neural crest-derived tumors that occur mostly sporadically, but may also be part of inherited syndromes. The molecular pathogenesis of sporadic pheochromocytomas remains unknown. Recently, the susceptibility gene for familial paraganglioma syndrome, a disorder embryologically related to pheochromocytomas, was characterized and shown to encode the small subunit of succinate dehydrogenase (SDHD), which is part of the mitochondrial complex II. This complex regulates oxygen-sensing signals. Importantly, hypoxic signals also appear to be related to the pathogenesis of pheochromocytomas associated with von Hippel-Lindau syndrome. We sequenced the entire coding region of the SDHD gene in a series of pheochromocytomas. Although we did not find mutations in the gene, we identified a new intronic single nucleotide polymorphism in 15% of the samples (g.97739A-->G). We also confirmed the existence of a sequence highly homologous to the SDHD complementary DNA in chromosome 1p34--36, a region commonly deleted in pheochromocytomas. Full analysis of this sequence revealed a heterozygous single base substitution in 70% of our samples that was also present in the germline. This sequence does not appear to be transcribed and is probably a processed pseudogene. Therefore, despite its chromosomal location, it is unlikely that this sequence is a target of loss of heterozygosity in pheochromocytomas. In conclusion, mutations of the SDHD gene are not a common event in this series of sporadic pheochromocytomas. The existence of SDHD pseudogenes should be considered when analyzing complementary DNA-based samples. PMID- 11397906 TI - Naturally occurring mutations in the melanocortin receptor 3 gene are not associated with type 2 diabetes mellitus in French Caucasians. AB - Familial genetic studies of type 2 diabetes (T2DM) of different human populations, including the French Caucasians, suggested evidence for linkage of T2DM and human chromosome 20q13, a region where maps the melanocortin 3 receptor gene (MC3R). Likewise, its homologous MC4R in human obesity, MC3R gene is also a good candidate for genetic susceptibility to glucose intolerance and T2DM. We therefore undertook a molecular study to assess the role of genetic variations of this gene in a large cohort of French families with T2DM. In these patients, we identified two missense mutations in the MC3R gene: Val(81)Ile and Lys(6)Thr. These two variants, which were in complete linkage disequilibrium, were also present in nondiabetic controls. Based on association and familial linkage disequilibrium tests results, we found that these MC3R gene-coding variants were not associated with diabetes or obesity. These variants were found, however, marginally associated with insulin and glucose levels during oral glucose tolerance testing in normoglycemic subjects. Overall, the present study provides no evidence for a major role of the MC3R coding mutations underlying the genetic linkages of T2DM and the MC3R gene region on chromosome 20q13 in T2DM families from France and other geographical origins. PMID- 11397907 TI - Inhibitory effect of a two day fast on reactive oxygen species (ROS) generation by leucocytes and plasma ortho-tyrosine and meta-tyrosine concentrations. AB - ABSTRACT Since glucose intake acutely increases reactive oxygen species (ROS) generation by polymorphonuclear leucocytes (PMN) and mononuclear cells (MNC), we have now investigated whether a fast over a period of 48h reduces ROS generation by these cells. Eight normal subjects were fasted for 48h. Blood samples were obtained at 0, 24h and 48h. ROS generation by PMN fell significantly at 24h (66.1 +/- 19.5% of basal) and further at 48h (45.9 +/- 23.0 % of basal; p < 0.001). ROS generation by MNC fell to 62.4 +/- 16.5% at 24h and by 48.4 +/- 16.5% (p < 0.001) by 48h. The level of p47(phox) subunit, an index of NADPH oxidase, the enzyme converting molecular oxygen to superoxide (O(.)(2)(-)) radical, also fell in parallel. Plasma o-tyrosine/phenylalanine ratio fell significantly from 0.326 +/- 0.053 mmol/mol to 0.303 +/- 0.055 mmol/mol at 48h and m-tyrosine/phenylalanine ratio fell from 0.363 +/- 0.063 mmol/mol to 0.340 +/- 0.064 mmol/mol (p < 0.05). Thus, a 48h fast may reduce ROS generation, total oxidative load and oxidative damage to amino acids. PMID- 11397908 TI - An extraordinarily inaccurate assay for free testosterone is still with us. PMID- 11397910 TI - Femtomolar sensitivity of metalloregulatory proteins controlling zinc homeostasis. AB - Intracellular zinc is thought to be available in a cytosolic pool of free or loosely bound Zn(II) ions in the micromolar to picomolar range. To test this, we determined the mechanism of zinc sensors that control metal uptake or export in Escherichia coli and calibrated their response against the thermodynamically defined free zinc concentration. Whereas the cellular zinc quota is millimolar, free Zn(II) concentrations that trigger transcription of zinc uptake or efflux machinery are femtomolar, or six orders of magnitude less than one atom per cell. This is not consistent with a cytosolic pool of free Zn(II) and suggests an extraordinary intracellular zinc-binding capacity. Thus, cells exert tight control over cytosolic metal concentrations, even for relatively low-toxicity metals such as zinc. PMID- 11397911 TI - Paleoclimate. Ice ages, the California Current, and Devils Hole. PMID- 11397912 TI - The role of Drosophila mushroom body signaling in olfactory memory. AB - The mushroom bodies of the Drosophila brain are important for olfactory learning and memory. To investigate the requirement for mushroom body signaling during the different phases of memory processing, we transiently inactivated neurotransmission through this region of the brain by expressing a temperature sensitive allele of the shibire dynamin guanosine triphosphatase, which is required for synaptic transmission. Inactivation of mushroom body signaling through alpha/beta neurons during different phases of memory processing revealed a requirement for mushroom body signaling during memory retrieval, but not during acquisition or consolidation. PMID- 11397913 TI - Multisensory integration and crossmodal attention effects in the human brain. PMID- 11397915 TI - Russian science. New rules on foreign contacts resurrect cold war-era distrust. PMID- 11397914 TI - Drug prices: real problem, wrong solution. PMID- 11397916 TI - Human rights. Academies seek release of Egyptian scientist. PMID- 11397917 TI - Embryonic stem cells. German leaders spar over bioethics. PMID- 11397918 TI - Space science. Canada eyes front-row seat in Mars program. PMID- 11397919 TI - Genetics. Faster maps mean fewer mice. PMID- 11397920 TI - PCR. Roche dealt a setback on European Taq patent. PMID- 11397921 TI - Astrobiology. Returning alien rocks right the second time. PMID- 11397922 TI - Smithsonian Institution. Director of Natural History Museum quits. PMID- 11397923 TI - Cancer research. Transatlantic war over BRCA1 patent. PMID- 11397924 TI - Paleoecology. Mass extinctions pinned on Ice Age hunters. PMID- 11397925 TI - Stem cell policy. Can adult stem cells suffice? PMID- 11397926 TI - Stem cell policy. An embryonic alternative. PMID- 11397927 TI - China's census. National count reveals major societal changes. PMID- 11397929 TI - Genetics. Why evolution might not favor increased genetic variability. PMID- 11397928 TI - Genetics. Can organisms speed their own evolution? PMID- 11397930 TI - Germany. A former capital stakes its future on science. PMID- 11397931 TI - Germany. Zoology institute shakes off the dust. PMID- 11397932 TI - Climate change. Fundamentals of treaty-making on climate change. PMID- 11397934 TI - Chemistry. Fleeting molecules extend their stay. PMID- 11397933 TI - Structural biology. A marvellous machine for making messages. PMID- 11397935 TI - Applied physics. Toward diamond lasers. PMID- 11397936 TI - Plate tectonics. Indian Ocean actively deforms. PMID- 11397937 TI - Molecular biology. Getting p53 out of the nucleus. PMID- 11397938 TI - Free-electron lasers. Status and applications. AB - A free-electron laser consists of an electron beam propagating through a periodic magnetic field. Today such lasers are used for research in materials science, chemical technology, biophysical science, medical applications, surface studies, and solid-state physics. Free-electron lasers with higher average power and shorter wavelengths are under development. Future applications range from industrial processing of materials to light sources for soft and hard x-rays. PMID- 11397939 TI - New ages for the last Australian megafauna: continent-wide extinction about 46,000 years ago. AB - All Australian land mammals, reptiles, and birds weighing more than 100 kilograms, and six of the seven genera with a body mass of 45 to 100 kilograms, perished in the late Quaternary. The timing and causes of these extinctions remain uncertain. We report burial ages for megafauna from 28 sites and infer extinction across the continent around 46,400 years ago (95% confidence interval, 51,200 to 39,800 years ago). Our results rule out extreme aridity at the Last Glacial Maximum as the cause of extinction, but not other climatic impacts; a "blitzkrieg" model of human-induced extinction; or an extended period of anthropogenic ecosystem disruption. PMID- 11397940 TI - A multispecies overkill simulation of the end-Pleistocene megafaunal mass extinction. AB - A computer simulation of North American end-Pleistocene human and large herbivore population dynamics correctly predicts the extinction or survival of 32 out of 41 prey species. Slow human population growth rates, random hunting, and low maximum hunting effort are assumed; additional parameters are based on published values. Predictions are close to observed values for overall extinction rates, human population densities, game consumption rates, and the temporal overlap of humans and extinct species. Results are robust to variation in unconstrained parameters. This fully mechanistic model accounts for megafaunal extinction without invoking climate change and secondary ecological effects. PMID- 11397941 TI - Room-temperature ultraviolet nanowire nanolasers. AB - Room-temperature ultraviolet lasing in semiconductor nanowire arrays has been demonstrated. The self-organized, <0001> oriented zinc oxide nanowires grown on sapphire substrates were synthesized with a simple vapor transport and condensation process. These wide band-gap semiconductor nanowires form natural laser cavities with diameters varying from 20 to 150 nanometers and lengths up to 10 micrometers. Under optical excitation, surface-emitting lasing action was observed at 385 nanometers, with an emission linewidth less than 0.3 nanometer. The chemical flexibility and the one-dimensionality of the nanowires make them ideal miniaturized laser light sources. These short-wavelength nanolasers could have myriad applications, including optical computing, information storage, and microanalysis. PMID- 11397942 TI - Ultraviolet emission from a diamond pn junction. AB - We report the realization of an ultraviolet light-emitting diode with the use of a diamond pn junction. The pn junction was formed from a boron-doped p-type diamond layer and phosphorus-doped n-type diamond layer grown epitaxially on the 111 surface of single crystalline diamond. The pn junction exhibited good diode characteristics, and at forward bias of about 20 volts strong ultraviolet light emission at 235 nanometers was observed and was attributed to free exciton recombination. PMID- 11397943 TI - (Amino)(aryl)carbenes: stable singlet carbenes featuring a spectator substituent. AB - Several (amino)(aryl)carbenes have been shown to be stable at room temperature in solution and in the solid state. Electroneutrality of the carbene center is ensured by the amino group, which has both pi-donor and final sigma-acceptor electronic character. The aryl group remains a spectator substituent, as shown by single-crystal x-ray analysis and by its chemical behavior. Because only one electron-active substituent is needed, numerous stable carbenes will become accessible, which will open the way for new synthetic developments and applications in various fields. PMID- 11397944 TI - Role of T-bet in commitment of TH1 cells before IL-12-dependent selection. AB - How cytokines control differentiation of helper T (TH) cells is controversial. We show that T-bet, without apparent assistance from interleukin 12 (IL-12)/STAT4, specifies TH1 effector fate by targeting chromatin remodeling to individual interferon-gamma (IFN-gamma) alleles and by inducing IL-12 receptor beta2 expression. Subsequently, it appears that IL-12/STAT4 serves two essential functions in the development of TH1 cells: as growth signal, inducing survival and cell division; and as trans-activator, prolonging IFN-gamma synthesis through a genetic interaction with the coactivator, CREB-binding protein. These results suggest that a cytokine does not simply induce TH fate choice but instead may act as an essential secondary stimulus that mediates selective survival of a lineage. PMID- 11397945 TI - A p53 amino-terminal nuclear export signal inhibited by DNA damage-induced phosphorylation. AB - The p53 protein is present in low amounts in normally growing cells and is activated in response to physiological insults. MDM2 regulates p53 either through inhibiting p53's transactivating function in the nucleus or by targeting p53 degradation in the cytoplasm. We identified a previously unknown nuclear export signal (NES) in the amino terminus of p53, spanning residues 11 to 27 and containing two serine residues phosphorylated after DNA damage, which was required for p53 nuclear export in colloboration with the carboxyl-terminal NES. Serine-15-phosphorylated p53 induced by ultraviolet irradiation was not exported. Thus, DNA damage-induced phosphorylation may achieve optimal p53 activation by inhibiting both MDM2 binding to, and the nuclear export of, p53. PMID- 11397946 TI - In silico mapping of complex disease-related traits in mice. AB - Experimental murine genetic models of complex human disease show great potential for understanding human disease pathogenesis. To reduce the time required for analysis of such models from many months down to milliseconds, a computational method for predicting chromosomal regions regulating phenotypic traits and a murine database of single nucleotide polymorphisms were developed. After entry of phenotypic information obtained from inbred mouse strains, the phenotypic and genotypic information is analyzed in silico to predict the chromosomal regions regulating the phenotypic trait. PMID- 11397947 TI - Guidelines for the referral and management of patients eligible for solid organ transplantation. AB - Members of the Clinical Practice Committee, American Society of Transplantation, have attempted to define referral criteria for solid organ transplantation. Work done by the Clinical Practice Committee does not represent the official position of the American Society of Transplantation. Recipients for solid organ transplantation are growing in numbers, progressively outstripping the availability of organ donors. As there may be discrepancies in referral practice and, therefore, inequity may exist in terms of access to transplantation, there needs to be uniformity about who should be referred to transplant centers so the system is fair for all patients. A review of the literature that is both generic and organ specific has been conducted so referring physicians can understand the criteria that make the patient a suitable potential transplant candidate. The psychosocial milieu that needs to be addressed is part of the transplant evaluation. Early intervention and evaluation appear to play a positive role in maximizing quality of life for the transplant recipient. There is evidence, especially in nephrology, that the majority of patients with progressive failure are referred to transplant centers at a late stage of disease. Evidence-based medicine forms the basis for medical decision-making about accepting the patient as a transplant candidate. The exact criteria for each organ are detailed. These guidelines reflect consensus opinions, synthesized by the authors after extensive literature review and reflecting the experience at their major transplant centers. These guidelines can be distributed by transplant centers to referring physicians, to aid them in understanding who is potentially an acceptable candidate for transplantation. The more familiar physicians are with the exact criteria for specific organ transplantation, the more likely they are to refer patients at an appropriate stage. Individual transplant centers will make final decisions on acceptability for transplantation based on specific patient factors. It is hoped that this overview will assist insurers/payors in reimbursing transplant centers for solid organ transplantation, based on criteria for acceptability by the transplant community. The selection and management of patients with end-stage organ failure are constantly changing, and future advances may make obsolete some of the criteria mentioned in the guidelines. Most importantly, these are intended to be guidelines, not rules. PMID- 11397948 TI - Protective role of tauroursodeoxycholate during harvesting and cold storage of human liver: a pilot study in transplant recipients. Transplantation 2001; 71: 1268. PMID- 11397949 TI - Assessment of Donor Fatty Livers for Liver Transplantation. Transplantation 2001; 71: 1221. PMID- 11397950 TI - Prolongation of sheep corneal allograft survival by ex vivo transfer of the gene encoding interleukin-10. Transplantation 2001; 71: 1214. PMID- 11397951 TI - Risk factors for failure to meet listing requirements in liver transplant candidates with alcoholic cirrhosis. AB - BACKGROUND: The majority of liver transplant centers require a 6-month abstinence period before listing candidates for liver transplantation with alcoholic cirrhosis and a persistent sobriety thereafter. We attempted to identify risk factors for failure to comply with these requirements. METHODS: Ninety-nine consecutive patients with alcoholic cirrhosis were referred for liver transplant evaluation between September 1996 and May 1998. The mean age was 49 years, 74% were male, and 54% were hepatitis C virus positive. To be listed, patients had to meet the following requirements. All patients received extensive psychosocial evaluations and were frequently monitored with random urine and blood alcohol tests; patients found positive were excluded or removed from the liver transplant waiting list. Detailed patient information was entered into a computerized database, and 36 discreet variables were analyzed in relation to success (patient listed and remained on the list) or failure (not listed or removed from the list based on noncompliance). RESULTS: Forty-nine patients were successfully listed. Nineteen received a transplant, with a 95% 1-year patient and graft survival rate and 21% alcohol relapse rate after transplantation. Twenty-two patients had either medical contraindication and/or died before transplant listing. Twenty four patients were never listed and four were removed from the list due to recurrent alcoholism, for a total of 28 failures. Our statistical analysis identified five significant risk factors for failure: (I) living arrangement (alone/family versus community/friend), P=0.006; (II) history of suicide ideation, P=0.03; (III) history of previous alcohol-related hospitalization, P=0.01; (IV) lack of previous alcoholic rehabilitation before transplant evaluation, P=0.001; and (V) failure to accept further alcoholic rehabilitation before orthotopic liver transplantation, P=0.01. CONCLUSIONS: Our experience confirms that transplantation can be extremely successful in properly selected patients with alcoholic cirrhosis. We identified several predictive psychosocial factors of early alcoholic recidivism in transplant candidates. PMID- 11397952 TI - Prolongation of sheep corneal allograft survival by ex vivo transfer of the gene encoding interleukin-10. AB - BACKGROUND: Modification of a donor cornea by gene therapy ex vivo has potential to modulate irreversible rejection, the major cause of corneal graft failure. Our aim was to transfer the gene encoding mammalian IL-10 to ovine donor corneas and to determine subsequent orthotopic corneal allograft survival in an outbred sheep model. METHODS: The replicative capacity of ovine corneal endothelium was determined by autoradiography after deliberate injury. A replication-defective adenovirus was used to deliver the lacZ reporter gene to ovine corneas and transfected corneas were organ-cultured in vitro to allow transfection efficiency, duration of reporter gene expression, and toxicity attributable to the vector to be determined. A cDNA encoding full-length ovine IL-10 was cloned into an adenoviral vector that was used to transfect donor corneas ex vivo before transplantation. Orthotopic penetrating corneal transplantation was performed in outbred sheep. RESULTS: Sheep corneal endothelium was found to be essentially amitotic. Transfection of > 70% corneal endothelial cells was achieved with the viral vector and expression was maintained for 28 days in vitro. IL-10 mRNA was detectable in transfected, organ-cultured corneas for 21 days in vitro. Donor corneas transfected with cDNA encoding IL-10 showed significantly prolonged survival after penetrating keratoplasty (median 55 days, range 19 > or =300 days) compared with control corneas (median 20.5 days, range 18-32 days, P=0.011). CONCLUSION: Local gene therapy-mediated expression of the immunomodulatory cytokine IL-10 has the potential to reduce the incidence of corneal graft rejection and to prolong corneal allograft survival. PMID- 11397953 TI - Assessment of donor fatty livers for liver transplantation. AB - AIM: The effect of fatty liver on graft survival, especially with reference to macrovesicular and microvesicular steatosis, is still uncertain. This preliminarily study was designed to create a noninvasive method for the quantification of the hepatic fat content in vivo and to establish provisional criteria for the assessment of fatty donor livers before liver transplantation among transplant surgeons, radiologists, and pathologists. METHODS AND MATERIALS: Different degrees of rat fatty liver model were established by feeding rats a diet deficient in choline and methionine for different periods of time. Computed tomography (CT) with test tubes containing variable percentages of fat equivalent substance were used to assess the severity of fatty change of the rat liver. This was then correlated with the histological classification, level of hepatic enzymes, and graft survival. RESULTS: Linear correlation between the fat volume fraction added to the test tubes and CT density were found. The process of producing a fatty liver via diet alteration peaked at week 3. At this time hepatic enzymes, radiological fat content, and posttransplantation survival were worse (P=0.013), compared with other time points. Radiological assessment of fatty liver correlated well with survival and serum glutamic oxaloacetic transaminase and glutamic pyruvate transaminase levels. CONCLUSION: Severe microvesicular steatosis does not influence recipient survival, however, macrovesicular steatosis affects graft survival. Caliber CT is a practical and simple method that allows an accurate noninvasive quantitative assessment of hepatic fatty infiltration. It has potential to be a useful parameter for the assessment of donor livers for clinical liver transplantation. PMID- 11397954 TI - Hepatocyte transplantation using biodegradable matrices in ascorbic acid deficient rats: comparison with heterotopically transplanted liver grafts. AB - BACKGROUND: Hepatocyte transplantation using polymeric matrices is under investigation as an alternative therapy for metabolic liver diseases. Long-term engraftment of hepatocytes in polymers has been demonstrated. However, the metabolic activity of hepatocytes in such devices has never been assessed in direct comparison with liver grafts. METHODS: Hepatocyte and partial liver transplantation were evaluated in the scurvy-prone osteogenic disorder Shionogi rat model. Biodegradable poly glycolic acid matrices seeded with hepatocytes equivalent to 20% of the recipient's liver mass, or 20% liver grafts were heterotopically transplanted into ascorbic acid- (AsA) deficient recipients. Recipients of cell-free matrices or AsA-deficient liver grafts served as controls. Recipients were set on AsA-free diet after transplantation. Plasma AsA levels, AsA concentrations in liver and adrenal gland tissue, and body weight ratios were assessed and H&E histology was performed. RESULTS: Recipients from the control groups showed symptoms of scurvy at 1 month after cessation of AsA supply. Hepatocyte transplantation and auxiliary liver transplantation prevented symptoms of scurvy and increased plasma and tissue AsA levels and body weight ratios. AsA levels in recipients of 20% liver grafts were comparable to normal control animals. CONCLUSIONS: Hepatocytes transplanted in polymeric matrices are able to compensate for liver-based metabolic deficiencies. Hepatocyte transplantation improves plasma AsA levels in AsA-deficient recipients. However, auxiliary liver grafts are superior to hepatocyte grafts in improving metabolic parameters. Further research work is needed to increase the efficiency of liver cell transplantation with regard to a clinical application. PMID- 11397955 TI - Hepatic xanthine levels as viability predictor of livers procured from non-heart beating donor pigs. AB - BACKGROUND: The aim of the present study was to evaluate hepatic content of adenine nucleotides and their degradation products in non-heart-beating donor (NHBD) pigs and its relationship with recipient survival. METHODS: Thirty animals were transplanted with an allograft from NHBDs. After warm ischemia (WI) time (20, 30, or 40 min), cardiopulmonary bypass and normothermic recirculation (NR) were run for 30 min. Afterward, the animals were cooled to 15 degrees C and liver procurement was performed. RESULTS: Survival rate was 100% in the 20WI, 70% in the 30WI, and 50% in the 40WI. Livers from non-surviving animals had higher levels of xanthine after NR than livers from surviving animals. Logistic regression analysis revealed that xanthine at the end of NR was the only variable able to predict survival with a calculated sensitivity of 80% and a specificity of 60%. Prolongation of warm ischemic period leaded to a greater xanthine accumulation as well as increased plasma alpha-glutathione S-transferase levels at reperfusion. Xanthine at NR and alpha-glutathione S-transferase at reperfusion significantly correlated, indicating that donor xanthine contributes to some extent to the severity of the lesion by ischemia-reperfusion. CONCLUSIONS: It is suggested that xanthine content in the donor is able to predict survival after transplantation. Xanthine is significantly involved in the hepatic lesion elicited by warm ischemia and subsequent ischemia-reperfusion associated to liver transplantation from a NHBD. PMID- 11397956 TI - Combined exogenous surfactant and inhaled nitric oxide therapy for lung ischemia reperfusion injury in minipigs. AB - BACKGROUND: The combined application of exogenous surfactant and inhaled nitric oxide was evaluated for prevention of ischemia-reperfusion injury of the lung. METHODS: Left lungs were selectively perfused in 18 minipigs in situ with cold preservation solution. After 90 min of warm ischemia, the lungs were reperfused and the right pulmonary artery and bronchus were ligated (control group, n=6). Exogenous surfactant was instilled via bronchoscopy during ischemia (surfactant group, n=6). In a third group, surfactant was applied, followed by administration of inhaled nitric oxide (surfactant+NO group, n=6). Hemodynamic and respiratory parameters were recorded for 7 hr, and bronchoalveolar lavage fluid (BALF) was obtained before and after reperfusion for measurement of surface tension, small aggregate/large aggregate ratio, protein and phospholipid contents, and a differential cell count. RESULTS: Control group animals survived for 3.7+/-1.4 hr. In both surfactant-treated groups, five out of six animals survived the observation period (P<0.001). Dynamic compliance of the lung was decreased in control animals (P<0.001). In the surfactant+NO group, arterial PO2 was higher than in both other groups (P<0.001). BALF cell count and histology showed reduced neutrophil infiltration in surfactant+NO-treated lungs. Surface tension assessed in BALF with a pulsating bubble surfactometer was severely impaired in control animals (gammamin, 14.82+/-9.95 mN/m), but maintained in surfactant-treated (gammamin, 1.11+/-0.56 mN/m) and surfactant+NO-treated animals (gammamin, 3.90+/ 2.35 mN/m, P=0.02). CONCLUSIONS: Administration of exogenous surfactant in lung reperfusion injury results in improved lung compliance. The addition of inhaled NO improves arterial oxygenation and reduces neutrophil extravasation compared with surfactant treatment alone. PMID- 11397957 TI - A novel paracorporeal method for isolated rodent lung reperfusion. AB - BACKGROUND: The isolated perfused lung model is commonly used in small animals to study lung function after preservation and cold storage. Measurements of oxygenation, compliance, and capillary filtration coefficient (Kf) permit analysis of preservation solutions or modifications of these solutions. However, inter-investigator variability using different perfusates makes comparisons difficult. Whole blood perfusion more closely mimics the in vivo situation, but extracorporeal circulation may alter the physiologic integrity of the model. Paracorporeal support has been used, but this technique required mechanical ventilation of the support rodent and did not incorporate a method for determining Kf. We evaluated a less-invasive technique, of providing cross circulatory syngeneic support, maintaining the ability to compute Kf. METHODS: Angiocatheters were inserted into both femoral arteries and one femoral vein of the support rat. The venous cannula was connected to the pulmonary artery of the ex vivo lung block to provide inflow. Pulmonary effluent blood from the lung block was collected via a left atrial cannula and returned to the support rat via the femoral artery. A separate, height-adjustable column was included in the circuit for measurement of Kf. RESULTS: Each support rat was used to sequentially perfuse three double-lung blocks. The inflow sample to each lung block was analyzed for pH, pO2, pCO2, and hematocrit to follow alterations in support rat physiology. There were no statistical differences in the pH, PO2, or hematocrit. No significant differences were noted in the pO2 of the pulmonary effluent blood or the Kf; analyzed to determine whether the sequence of reperfusion affected the pulmonary function assessment. CONCLUSIONS: The syngeneic support rat delivers constant pressure systemic venous blood at stable physiologic parameters to the ex vivo lung block. Recirculation of the perfusate through the support rat diminishes the need to pool blood from donors, detoxifies and deoxygenates pulmonary effluent blood, and permits examination of sequential lung blocks. This technique represents a hybrid model between isolated perfused and orthotopic transplant models, maintaining Kf determination, a sensitive indicator of reperfusion injury. This technique could be applicable to reperfusion injury models of other organs (using arterial inflow instead) and may permit increased standardization among investigators. PMID- 11397958 TI - Orthogonal polarization spectral imaging as a tool for the assessment of hepatic microcirculation: a validation study. AB - BACKGROUND: Quantitative analysis of liver microcirculation using intravital fluorescence microscopy in animals has increased our knowledge about ischemia reperfusion injury. However, because of the size of the instrumentation and the necessity of fluochromes for contrast enhancement, human liver microcirculation cannot be observed. Orthogonal Polarization Spectral (OPS) imaging is a recently introduced technique that can be used to visualize the microcirculation without the need for fluorescent dyes. It is a small, hand-held device and could potentially be used to study the microcirculation of the human liver in a clinical setting. However, before implementation into clinical use its ability to quantitatively measure microcirculatory parameters must be validated. METHODS: The livers of Spraque-Dawley rats (n=9) were exteriorized, and images were obtained using OPS imaging and intravital fluorescence microscopy of the identical microvascular regions before and after the induction of a 20-min warm lobar ischemia. Images were videotaped for later computer-assisted off-line analysis. RESULTS: OPS imaging can be used to accurately quantify the sinusoidal perfusion rate, vessel diameter, and venular red blood cell velocity. Correlation parameters were significant and Bland-Altman analyses showed good agreement for data obtained from the two methods at baseline as well as during reperfusion. CONCLUSION: OPS imaging can be used to quantitatively measure microcirculatory parameters in the rat liver under both physiological and pathophysiological conditions. Thus, OPS imaging has the potential to be used to make quantitative measurements of the microcirculation in the human liver. PMID- 11397959 TI - Cytomegalovirus infection, viral DNA, and immediate early-1 gene expression in rejecting rat liver allografts. AB - BACKGROUND: Cytomegalovirus (CMV) infection has been linked to acute and chronic rejection. We have previously shown that concomitant rat cytomegalovirus (RCMV) infection increases portal inflammation and bile duct destruction in rejecting rat liver allografts. Many of the pro-inflammatory effects of CMV have been attributed to the immediate early (IE) proteins of CMV. We wanted to investigate whether RCMV and IE-1 gene expression persist in the liver graft in our model. METHODS: Liver transplantations were performed from PVG (RT1c) into BN (RT1n) rats. One day after transplantation, the rats were infected with RCMV. No immunosuppression was given. The graft infection was studied by viral culture, immunofluorescence, DNA in situ hybridization and RT-PCR for the detection of IE 1 mRNA at various time points. RESULTS: RCMV caused an active infection from 5 days to 2 weeks after transplantation, during which infectious virus was found in the graft. Thereafter the cultures were negative. RCMV antigens and DNA were found in hepatocytes, endothelial, inflammatory, and bile duct cells during the active infection. At 4 weeks, RCMV DNA positive hepatocytes, endothelial, inflamma tory, and bile duct cells could still be found, but in much smaller quantities. IE-1 mRNA expression was, however, only detected during the active infection, not at 4 weeks postinfection. CONCLUSIONS: RCMV IE-1 expression does not persist in the graft after the active infection, although some viral DNA can be detected in the graft up to 4 weeks. In our model, the CMV-induced increase in graft damage does not seem to require the continued expression of IE-1. PMID- 11397960 TI - Inhibition of interferon-gamma-mediated microvascular endothelial cell major histocompatibility complex class II gene activation by HMG-CoA reductase inhibitors. AB - BACKGROUND: Graft vascular disease, a major cause of late graft failure in cardiac transplant patients, is associated with the presence of class II major histocompatibility complex molecules on the endothelium. 3-hydroxy-3 methylglutaryl (HMG)-CoA reductase inhibitors, e.g., simvastatin, have been shown to reduce the incidence of graft vascular disease. We studied the effect of simvastatin on interferon (IFN)-gamma-induced human leukocyte antigen (HLA)-DR expression in human microvascular endothelial cells (MVECs). METHODS AND RESULTS: Simvastatin pretreatment inhibited MVEC HILA-DR induction by IFN-gamma, as detected by flow cytometry. Simvastatin's inhibitory effect was reversed by the cholesterol synthesis pathway intermediates mevalonate and geranylgeranyl pyrophosphate but not squalene, indicating the involvement of protein prenylation in this process. Reverse transcription-polymerase chain reaction analysis demonstrated that induction of class II transactivator (CIITA), and consequently, HLA-DRalpha mRNA, is abrogated by simvastatin. Although signal transducer and activator of transcription (STAT)-1 is a critical CIITA gene transactivator, immunofluorescence studies, Western blotting, and electrophoretic mobility shift assays demonstrated that IFN-gamma-induced STAT-1 phosphorylation, nuclear translocation, and DNA binding are not affected by simvastatin. However, simvastatin inhibited IFN-gamma-induced transactivation of a CIITA promoter IV reporter construct, indicating the involvement of this promoter in the inhibitory effect of simvastatin. CONCLUSIONS: Simvastatin pretreatment inhibits CIITA and consequent HLA-DR induction by IFN-gamma in MVECs through interference with protein prenylation. This inhibitory effect occurs at the level of transcription and is directed, at least in part, at the CIITA promoter IV. These results explain some of the beneficial effects of HMG-CoA reductase inhibitors in cardiac transplantation. PMID- 11397961 TI - Protective role of tauroursodeoxycholate during harvesting and cold storage of human liver: a pilot study in transplant recipients. AB - BACKGROUND: Ischemia-reperfusion injury is a major cause of early graft dysfunction after liver transplantation. Tauroursodeoxycholic acid (TUDCA), a natural amidated hydrophilic bile salt, protects from cholestasis and hepatocellular damage in a variety of experimental models, as well as from ischemia-reperfusion injury. We investigated in the human liver transplantation setting the effect of the addition of TUDCA at time of liver harvesting and cold storage on the intra- and postoperative enzyme release and liver histopathology at the end of cold storage, at reperfusion, and 7 days after transplantation. METHODS: Eighteen patients undergoing elective liver transplantation were studied, including 6 serving as controls. In six patients, TUDCA was added to the University of Wisconsin solution used during harvesting and cold storage, to reach final concentrations of 2 mM. In three of these patients, TUDCA (3 g) was infused in the portal vein of the donor before organ explantation; in the other three cases, TUDCA was given through both routes. RESULTS: The use of TUDCA did not cause adverse events. The release of aspartate aminotransferase in the inferior vena cava blood during liver flushing was significantly lower (P=0.05) in TUDCA-treated than in control grafts, as were cytolytic enzyme levels in peripheral blood during the first postoperative week (P<0.02). At electron microscopy, an overt endothelial damage (cytoplasmic vacuolization, cell leakage, and destruction with exposure of hepatocytes to the sinusoidal lumen) was invariably found in control grafts, both at reperfusion and at day 7 after transplant. These features were significantly ameliorated by TUDCA (P<0.001). Several ultrastructural cytoplasmic abnormalities of hepatocytes were seen. Among these, damage to mitochondria matrix and crystae was significantly reduced in TUDCA-treated versus control grafts (P<0.01). Mild to severe damage of bile canaliculi was a constant feature in control biopsies, with dilatation of canalicular lumen and loss of microvilli. Both these abnormalities were markedly ameliorated (P<0.001 by TUDCA). The best preservation was observed when TUDCA was given through both routes. CONCLUSIONS: The use of TUDCA during harvesting and cold storage of human liver is associated with significant protection from ischemia-reperfusion injury. The clinical significance of this findings must be studied. PMID- 11397962 TI - Autonomic dysfunction and ambulatory blood pressure in renal transplant recipients. AB - BACKGROUND: Abnormal circadian blood pressure (BP) rhythm (nondipping) and autonomic dysfunction are both common in end-stage renal disease (ESRD). It is not known whether these abnormalities are related or if they are associated with greater left ventricular hypertrophy. METHODS: Nineteen renal transplantation (RT) recipients, aged 22-67 years, who were transplanted at least 12 months (1-29 years) previously, were studied with 24-hr ambulatory blood pressure monitoring (ABPM). Autonomic function was tested by automated analysis of heart rate variations and echocardiography was used to estimate left ventricular mass index (LVMI). RESULTS: Thirteen patients (68%) were nondippers. Although seven (37%) patients had significant parasympathetic dysfunction, this was not related to dipper status. Neither abnormality showed a tendency to diminish with time after RT. Systolic hypertension, diagnosed by ABPM, occurred in 5% of patients during the awake period and in 52% during sleep, whereas diastolic hypertension occurred in 47% when awake and in 63% when asleep. Awake systolic BP was the strongest predictor of LVMI (r=0.7, P<0.001), and was considerably better than systolic BPs recorded at the clinic (r=0.48, P<0.05). CONCLUSIONS: Nondipping is common after RT but is not related to the degree of autonomic dysfunction. These findings suggest that autonomic dysfunction is not a major contributor to nondipping in ESRD. In RT patients, ABPM is a more sensitive measure of hypertension and a stronger predictor of LVMI than clinic BP. PMID- 11397964 TI - Levels of virus-specific CD4 T cells correlate with cytomegalovirus control and predict virus-induced disease after renal transplantation. AB - BACKGROUND: Immunosuppressive treatment in transplant patients frequently causes infectious complications with cytomegalovirus (CMV). The extent of CMV replication can be followed by a number of diagnostic methods. There is, however, no simple diagnostic tool to assess the quality of the cellular antiviral immune response of an individual patient. This would be of particular importance for therapy decisions, as patients with detectable virus load do not necessarily develop CMV-related disease. Using a rapid whole blood assay, the frequencies of CMV-reactive CD4 and CD8 T cells were followed after renal transplantation to characterize their relative contribution in the containment of CMV infection. METHODS: T cells from transplant patients ands healthy control persons were stimulated with CMV antigen in vitro. Based on specific cellular activation and induction of intracellular cytokines, the frequency of CMV-reactive CD4 and CD8 T cells was determined using flow cytometry. Viral load quantified using the "hybrid-capture" assay. RESULTS: The absence of CMV complications in long-term transplant recipients is reflected by stable virus-specific T-cell frequencies, which do not differ from healthy CMV-positive controls. In contrast, during the first months after transplantation, clinical symptoms are preceded by a decrease in CMV-reactive CD4 T-cell frequencies and an increase in CMV load. CONCLUSIONS: The individual immune response and CMV replication are critically balanced and can be characterized by assesing both viral load and antiviral T cells. Our experimental design allows the identification of patients with sufficient, insufficient, or absent T-cell activity and can serve as diagnostic tool to facilitate decisions on antiviral therapy. PMID- 11397963 TI - Multicenter trial exploring calcineurin inhibitors avoidance in renal transplantation. AB - BACKGROUND: The adoption of calcineurin inhibitors (CNI) as the mainstay of immunosuppression has resuited in a significant decrease of acute rejection and improvement of short-term graft survival. However, because of the irreversible nephrotoxicity associated with the chronic use of the CNI, the magnitude of the improvement of long-term graft survival has been more modest. Therefore, an effective immunosuppression regimen that does not rely on CNI may result in improvement of long-term outcome and simplification of the management of transplant recipients. METHODS: Ninety-eight patients of primary cadaver or living donor kidneys at low immunologic risk were enrolled in a CNI avoidance study. The immunosuppression regimen consisted of daclizumab, a humanized monoclonal antibody that binds to the alpha chain of the interleukin-2 receptor (IL-2Ralpha), administered for a total of five doses at biweekly intervals; 3 gm/day mycophenolate mofetil for the first 6 month and 2 gm thereafter; and conventional corticosteroid therapy. Patients who underwent rejection episodes could be started on CNI. The primary efficacy end-point was biopsy-proven rejection during the first 6 months posttransplant. RESULTS: Biopsy-proven rejection was diagnosed in 48% of patients during the first 6 months after transplantation. The majority of rejection episodes were Banff grade I and IIA and were fully reversed with corticosteroid therapy. The median time to the first biopsy-proven rejection among patients who experienced this event during the first 6 months was 39 days. In 22 patients with delayed graft function, the proportion of patients with biopsy-proven rejection was 50% at 6 months. However in the first 2 weeks posttransplant, only 1 of 22 patients with delayed graft function developed biopsy-proven rejection. At 1 year, patient survival was 97% and graft survival was 96%. Only two grafts were lost secondary to rejection. At 1-year posttransplant, 62% of patients had received CNI for more than 7 days. At 1-year posttransplant, the mean serum creatinine in the nonrejectors with no CNI use was 113 micromol/L (95%, confidence interval [CI], 100.7 to 125.3 micromol/L) and in the rejectors or patients with CNI use (more than 7 days) was 154 micromol/L (95% CI, 135.0 to 173.0 micromol/L). In selected patients with rejection, analysis of circulating and intragraft lymphocytes revealed complete IL-2Ralpha saturation. CONCLUSIONS: This CNI avoidance study in immunologic low risk patients, while only partially successful in preventing acute rejection, provided benefits to a sizable minority of patients who have not required chronic CNI therapy. However, wide acceptance of a CNI-sparing immunosuppression regimen may require a lower rate of acute rejection, possibly through the addition of a non-nephrotoxic dose of CNI. However, because complete IL-2Ralpha blockade was present during rejection, it can be assumed that alternative pathways, such as IL 15, may be responsible for the rejection; thus, the incorporation of non nephrotoxic immunosuppressive agents, such as sirolimus, may provide a more strategic approach. PMID- 11397966 TI - JC virus infection in allograft kidneys: analysis by polymerase chain reaction and immunohistochemistry. AB - BACKGROUND: Polyoma virus nephropathy after transplantation is believed to be primarily due to the BK virus. We hypothesized that some cases may be associated with the JC polyoma virus (JCV), which is also known to be latent in the kidney. METHODS: We sought polymerase chain reaction evidence of JCV infection in needle biopsy specimens with and without viral nephropathy. Cases positive by polymerase chain reaction were studied by immunohistochemistry for VP-1 antigen expression. RESULTS: JCV DNA was found in 7 (36.8%) of 19 allograft kidney biopsy specimens with viral nephropathy and 0 (0%) of 19 native or allograft biopsy specimens without viral nephropathy. Immunohistochemistry localized JCV to the nuclei of tubular epithelial cells in one case. CONCLUSIONS: JCV is detectable in a subset of renal allograft kidneys with polyoma virus nephropathy. The tubular epithelium is identified as a site capable of supporting JCV viral capsid protein VP-1 expression, and hence viral replication. PMID- 11397965 TI - Preventive effect of inhaled nitric oxide and pentoxifylline on ischemia/reperfusion injury after lung transplantation. AB - BACKGROUND: The preventive effect of inhaled nitric oxide (NO) and pentoxifylline (PTX) administered during reperfusion has been demonstrated on experimental models of lung ischemia/reperfusion (I/R) injury but this strategy is not validated in clinical lung transplantation. The aim of this study was to assess retrospectively the protective effect of inhaled NO and PTX after lung transplantation. METHODS: Twenty-three consecutive patients who received inhaled NO (10 ppm) and PTX (NO-PTX group) at the time of reperfusion were compared retrospectively with (1) 23 consecutive patients transplanted just before the use of NO-PTX (control group 23); (2) 95 patients representing all the patients of the series who did not receive NO-PTX (control group 95), with respect to I/R injury related complications. In particular, the incidence of pulmonary reimplantation edema and early hemodynamic failure, the PaO2/FIO2 ratio as well as the duration of mechanical ventilation and the 2-month mortality rates were compared. RESULTS: Reimplantation edema was observed in 6/23 patients (26%) in the NO-PTX group vs. 13/23 patients (56%) in the control group 23 (P=0.035) and 48/95 patients (50%) in the control group 95 (P=0.035). The worst PaO2/FIO2 ratio during the first three postoperative days was 240-102 mmHg in the NO-PTX group vs. 162+/-88 mmHg (P=0.01) and 176+/-107 mmHg (P=0.01) in the control group 23 and the control group 95, respectively. The duration of mechanical ventilation was 2.1+/-2.4 days in the NO-PTX group vs. 7+/-9 days in the control group 23 (P=0.02) and 6+/-7 days in the control group 95 (P=0.01). The 2-month mortality rate was 4.3% in the NO-PTX group vs. 26% (P=0.04) and 21% (P=0.07) in the control group 23 and the control group 95, respectively. CONCLUSIONS: The marked decrease in the incidence of allograft dysfunction compared with two historical control groups suggests that PTX and inhaled NO given before and throughout reperfusion are protective against I/R injury in the setting of clinical transplantation. PMID- 11397967 TI - The effect of gut metabolism on tacrolimus bioavailability in renal transplant recipients. AB - BACKGROUND: Tacrolimus, a substrate of CYP3A, has low and variable bioavailability similar to cyclosporine. Co-administration of ketoconazole, potent inhibitor of gut and hepatic CYP3A, has been shown to increase tacrolimus bioavailability in healthy volunteers. The purpose of this study is to assess the role of gut metabolism in the overall bioavailability of tacrolimus in a renal transplant population. METHODS: We prospectively studied 19 adult renal transplant recipients who were receiving tacrolimus as part of a quadruple, sequential immunosuppression regimen. Each patient received tacrolimus (4-hr intravenous dose of 0.04 mg/kg between postoperative days 2 and 4). Whole blood samples were collected over 24 hr. After a 24-hr washout period, a single oral dose of ketoconazole (400 mg) was administered followed by the same intravenous dose of tacrolimus, and subsequent samples were obtained. Steady state oral pharmacokinetic profiles were obtained between 1 and 3 months after transplant while patients were receiving twice daily dosing of tacrolimus to maintain whole blood levels between 10 and 20 ng/ml. Two days later, 400 mg of ketoconazole was administered orally 2 hr before to the morning dose. Whole blood samples were collected over 12 hr. RESULTS: In the absence of ketoconazole, 8.0% of the tacrolimus dose underwent first pass metabolism (E(H)), whereas in the presence of ketoconazole, first pass metabolism was 6.2% (P=0.01). Based on this difference in first pass metabolism, an increase of 2% in bioavailability is expected, but an increase of 47% is observed (P=0.001). CONCLUSIONS: This indicates that the gut metabolism of tacrolimus is extensive and contributes significantly to its bioavailability. PMID- 11397968 TI - Complement activation products in plasma after heart transplantation in humans. AB - BACKGROUND: Complement activation has recently been implicated as a contributing factor to early and late allograft dysfunction in cardiac transplantation. The current study was designed to determine whether measurement of plasma complement fragments C4d and SC5b-9 would be useful in detecting acute rejection or accelerated graft atherosclerosis (AGA) in cardiac allograft recipients. METHODS: We measured complement activation products, C4d (classical pathway) and SC5b-9 (terminal pathway), at the time of routine endomyocardial biopsy in heart transplant recipients. Ten patients in the immediate posttransplantation period (0-100 days) and 19 patients more than 6 months after transplantation were studied. RESULTS: No correlation was found between plasma levels of complement activation fragments and the presence of biopsy-proven acute allograft rejection or AGA (assessed by coronary angiography). However, plasma C4d and SC5b-9 were significantly elevated in 9 of 10 and 7 of 10 patients, respectively, in the immediate posttransplantation period. This was followed by progressive decrease in the levels of C4d and SC5b-9 fragments during the first 4-6 weeks after transplantation. CONCLUSION: We conclude that measuring plasma levels of fragments C4d and SC5b-9 is not a useful noninvasive method for detecting acute rejection or AGA after heart transplantation. However, this study provides further evidence that early complement activation after heart transplantation may play a pathogenic role in allograft injury. PMID- 11397969 TI - Guillain-Barre syndrome after solid organ transplantation. AB - BACKGROUND: Neurological complications occur frequently in solid organ transplant recipients. However, the peripheral nerves are usually spared significant toxicity. Guillain Barre syndrome (GBS) is the most common cause of acute neuropathy in adults. Despite numerous reports of GBS in recipients of bone marrow transplants, GBS has rarely been reported in recipients of solid organ transplants. Recent evidence supports the role of the immune system in initiating and perpetuating the ongoing neural damage in this entity. Infectious agents may initiate the immune attack, and the association of GBS with cytomegalovirus (CMV) infection has been studied extensively. METHODS: To alert clinicians to the occurrence of GBS in the latter setting, we report five new cases of GBS after solid organ transplant and summarize five other cases previously reported in the literature. RESULTS: The GBS cases (published and unpublished) have much in common: all the patients were men, most had evidence of active CMV infection at or before the onset of GBS, and all but one developed GBS within 1 year after transplantation (range 1-26 months). CONCLUSION: The association of GBS with cytomegalovirus (CMV) infection in the nontransplant population and evidence of CMV infection in almost all reported cases of GBS in solid organ transplant recipients suggest that CMV may have a role in triggering this illness. PMID- 11397970 TI - Serial evaluation of cell surface markers for immune activation after acute renal allograft rejection by urine flow cytometry--correlation with clinical outcome. AB - BACKGROUND: The use of urine flow cytometry (UFC) as a noninvasive tool for the diagnosis of acute and chronic rejection of the renal allograft has been previously reported. METHODS: We analyzed the expression of various cell surface antigens during a 30-day period after the diagnosis and treatment of 24 acute rejection (AR) episodes. UFC was performed on 59 urine specimens, from 17 patients meeting the diagnostic criteria for AR. UFC analysis was performed blinded to the clinical management utilizing the following fluorescinated monoclonal antibodies: anti-CD3, anti-CD14, anti-HLA-DR, anti-CD54, and anti interleukin 2 receptor. Results were correlated with the patient's requirement for antilymphocytic drugs and increment in serum creatinine level (mg/dl) on day 30 after AR. RESULTS: HLA-DR was the most prevalent antigen noted during the first 2 days of AR (91.7% of the samples), followed by CD14 (50%) and CD54 (41.7%). After day 4 the degree of expression of HLA-DR-, CD14-, and CD54 positive cells correlated with the need for antilymphocytic drugs. CD54 was the best parameter with a sensitivity=100% and specificity=90.9% (P=0.001). Those patients who had permanent graft injury after treatment of the AR had persistence of CD54- and CD14-positive cells in the urine. CONCLUSION: Serial monitoring of urine sediments by UFC was predictive of the requirement for antilymphocytic therapy and irreversible graft damage. PMID- 11397971 TI - A calcineurin antagonist-free induction strategy for immunosuppression in cadaveric kidney transplant recipients at risk for delayed graft function. AB - BACKGROUND: Avoidance of calcineurin antagonists for a prolonged period de novo after cadaver donor renal transplantation may facilitate recovery from delayed graft function. The present study examined the benefit of prolonging the calcineurin antagonist-free interval by administering sirolimus (SRL) in combination with chimeric (c-) anti-interleukin-2 receptor (IL-2R) monoclonal antibodies (mAb). METHODS: Three contemporaneous but nonrandomized cohorts were compared for acute rejection episodes, patient and graft survival rates, renal function, and adverse reaction profiles for 12 months. Patients with delayed graft function were treated with either SRL/c-IL-2R mAb/prednisone (Pred) with inception of cyclosporine (CsA) once the serum creatinine value was < or =2.5 mg/dl (n=43; group 1) or anti-lymphocyte preparations/Pred/delayed CsA for 7 to 14 days (n=18; group 3). A third cohort displayed immediate function and was treated de novo with CsA/c-IL-2R mAb/Pred (n=21; group 2). RESULTS: The incidence of acute rejection episodes was significantly lower among group 1 (16%) compared with groups 2 (52%, P=0.004) or 3 (39%, P=0.05). Among the seven rejection episodes in group 1, six of seven occurred among African-American or retransplant recipients, and a separate cluster of six of seven occurred among patients who displayed SRL trough concentrations < or =9 ng/ml. Furthermore, additional antilymphocyte antibody treatment was required to reverse either steroid resistant or Banff grades II or III acute rejection episodes among 14%, 55% (P=0.08), and 71% (P=0.03) of patients in each group, respectively. Patient and graft survival rates, as well as mean serum creatinine values, were similar at 12 months among the three groups. However, group 1 patients displayed higher serum cholesterol and triglyceride values, as well as lower hemoglobin, platelet, and leukocyte values compared with the other two groups. CONCLUSION: This pilot study suggests that a SRL/c-IL-2R mAb/Pred induction regimen provides excellent acute rejection prophylaxis. PMID- 11397972 TI - Early chimerism of macrophages and lymphocytes in lung transplant recipients is predictive of graft tolerance. AB - BACKGROUND: The persistence of donor cells derived from the graft (chimerism) has been documented in various tissues after organ transplantation. It was suggested that stable chimerism might reflect a state of donor-specific tolerance. Chimerism of macrophages and lymphocytes were studied over time after lung transplantation as well as its impact on graft tolerance. MATERIAL AND METHODS: Macrophages and lymphocytes were purified from bronchoalveolar lavage sequentially obtained from 24 patients between 1 and 41 months posttransplantation (20, 22, 24, and 17 patients at, respectively, 1, 3, 6, and 12 months). DNA was extracted from these cells and their recipient-donor origin was evaluated by PCR amplification of highly polymorphic DNA regions (minisatellites). RESULTS: We show that the remaining donor cells over the first month vary from 10 up to 50% and 5 up to 55% for lymphocytes and macrophages respectively (+/-2 SD). All patients presented some chimerism up to the 6th postoperative month. Good correlation was observed between the residual amount of donor lymphocytes and macrophages during the first 3 months (P<0.001). Patients with at least 30% donor lymphocytes at 1 month after transplantation had less rejections (> or =stage II) in the follow up (P=0.0007). The same observation is true for donor macrophages although to lower extend (P=0.02). The chimerism lost its predictive value beyond 3 months. CONCLUSIONS: These data demonstrate that a level of chimerism above 30% of either donor lymphocytes or macrophages at 1 month is related to a better state of graft tolerance. However, chimerism decreases markedly beyond 3 months and has then no predictive value. PMID- 11397973 TI - Differential effects of myeloid dendritic cells retrovirally transduced to express mammalian or viral interleukin-10 on cytotoxic T lymphocyte and natural killer cell functions and resistance to tumor growth. AB - BACKGROUND: Genetic engineering of dendritic cells (DC) to express immunosuppressive molecule(s) offers potential for therapy of allograft rejection and autoimmune disease. Viral (v) interleukin (IL)-10, encoded by the Epstein Barr virus, is highly homologous to mammalian (m) IL-10, but lacks certain of its T-cell stimulatory activities. Our aim was to evaluate and compare the influence of vIL-10 and mIL-10 gene transfer on the T-cell and natural killer (NK) cell stimulatory activity of DC, and their impact on the growth of transplantable tumors. METHODS: Myeloid DC progenitors, propagated from the bone marrow of C57BL/6J (H2b) mice in granulocyte-macrophage colony-stimulating factor + IL-4, were transduced using retroviral supematant from the BOSC ecotropic packaging cell line. The function of the IL-b gene-modified DC was assessed by examining their ability to induce naive allogeneic T-cell proliferation and cytotoxic T lymphocyte (CTL) generation. MCA205 (H2b) sarcoma cells mixed with either vIL-10 , mIL-10-, or Zeo (control gene)-transduced DC were inoculated intradermally into C57BL/6J (syngeneic) or BALB/cJ (H2d) (allogeneic) recipients, which were monitored for tumor growth. The role of specific host effector cell populations in tumor resistance was determined by antibody depletion. RESULTS: Compared with control gene-modified DC, both vIL-10- and mIL-10-transduced DC, which secreted the transgene product, showed reduced surface expression of MHC class II and costimulatory molecules, and impaired ability to induce T-cell proliferation. vIL 10-transduced DC were also inhibited with respect to CTL induction but did not affect the generation of NK cells. By contrast, mIL-10-transduced DC augmented CTL generation and NK cell activity. In the tumor transplant model, vIL-10 transduced DC enhanced tumor growth both in syngeneic and allogeneic hosts, whereas mIL-10-transduced cells inhibited tumor development. Depletion of CD4+ or CD8+ T cells or NK cells in mice given mIL-10-transduced DC reversed this therapeutic effect. CONCLUSION: mIL-10 gene-modified myeloid DC promote CTL and NK cell-mediated responses and inhibit tumor growth. By contrast, vIL-10 engineered DC, which elicit diminished CTL responses and do not promote NK cell activity, seem to have therapeutic potential for inhibition of T cell-mediated immunity. PMID- 11397974 TI - Successful lung transplantation for bronchiolitis obliterans after allogeneic marrow transplantation. AB - BACKGROUND: Bone marrow transplantation (BMT) is an established therapy for a variety of hematological diseases with curative potential. However, despite improvements in supportive care, pulmonary complications remain a significant cause of morbidity and mortality. METHODS: We report on a patient who received a double lung transplantation (LTX) for therapy-refractory bronchiolitis obliterans (BO) associated with extensive chronic graft-versus-host disease (GVHD) after allogeneic BMT. RESULTS: At present, 38 months after BMT and 23 months after LTX, the patient is in complete hematological and cytogenetic remission and without signs of respiratory distress. CONCLUSIONS: This case illustrates that lung transplantation could be a therapeutic option in selected patients with BO after allogeneic BMT that is associated with extensive chronic GVHD and who are refractory to conventional immunosuppressive therapy. PMID- 11397975 TI - Cerebral phaeohyphomycosis due to a novel species: report of a case and review of the literature. AB - Cerebral phaeohyphomycosis is a rare disease caused by dematiaceous (darkly pigmented) fungi. Cladophialophora species are highly neurotropic, and Cladophialophora bantiana (synonym=Xylohypha bantiana or C. trichoides) is the most commonly identified agent. Most reported cases of cerebral phaeohyphomycosis have occurred in immunocompetent patients; however, some case reports and experimental data have suggested that cellular immune deficiency is a risk factor. We report a case of pulmonary and cerebral phaeohyphomycosis in a cardiac transplant patient due to a newly identified species of Cladophialophora. Optimal management includes both antifungal therapy and surgery. PMID- 11397976 TI - Domino hepatic transplantation using the liver from a patient with primary hyperoxaluria. AB - BACKGROUND: We report a case of domino liver transplantation using the liver harvested from a patient who underwent a combined liver and kidney transplantation for primary hyperoxaluria (PH). METHOD: A cadaveric liver transplantation was performed in a 19-year-old man with PH. In a second step, the PH liver harvested from the first patient was transplanted in a 69-year-old man with hepatitis C-related cirrhosis, not a candidate for a classic liver graft owing to multifocal hepatocellular carcinoma. RESULTS: At 8 months after transplantation, the domino recipient has normal hepatic function and no signs of tumoral recurrence, but he progressively developed hyperoxalemia, hyperoxaluria, and renal insufficiency. CONCLUSION: Regarding the favorable postoperative clinical evolution, domino liver transplantations using livers from PH patients may represent a new opportunity for marginal candidates for liver transplantation. However, the progressive renal insufficiency expected in such domino recipients should limit this procedure to selected cases. PMID- 11397977 TI - Renal cell carcinoma detected in a cadaveric donor after orthotopic liver and contralateral renal transplantation in two recipients: four-year follow-up. AB - BACKGROUND: Although rare, renal cell carcinoma has been found during renal recovery for cadaveric organ transplantation. Previously, we reported this incidence to be 0.9%. In one cadaveric donor, the liver and left kidney had been transplanted before the discovery of renal cell carcinoma (T1) in the right kidney. METHODS: We retrospectively reviewed the medical records of two patients who had received cadaveric allografts from a donor with a known renal cell carcinoma. RESULTS: Both patients have been followed for 4 years with blood chemistries and chest x-ray every 3 months for year 1, every 4 months for years 2 and 3, and every 6 months thereafter. They also underwent allograft ultrasound every 6 months and an annual CT scan of the abdomen. Both patients have shown no evidence of metastatic disease throughout their follow-up. DISCUSSION: In the rare instance that a patient receives an organ from a cadaveric donor with a known renal cell carcinoma, it is mandatory to follow these patients closely observing for both allograft recurrence and metastatic disease. PMID- 11397978 TI - Laparoscopic right adrenalectomy after liver transplantation. AB - To our knowledge, laparoscopic right adrenalectomy has not been previously reported after orthotopic liver transplantation. The aim of this report is to demonstrate the feasibility of the laparoscopic approach in this technically challenging situation, and to outline some considerations unique to this clinical setting. PMID- 11397979 TI - Strategies for meaningful measurement of circulating levels of transforming growth factor beta1 in clinical samples. PMID- 11397980 TI - [Etiologies of ocular and eyelid trauma in Burkina Faso]. AB - PURPOSE: To identify the epidemiological features of ocular traumas in Burkina Faso. METHODS: A retrospective study was conducted in the eye department of the National Hospital of Ouagadougou, from January 1995 to 31 December 1997. All cases of eye trauma during this period were included. RESULTS: The study showed that ocular traumas represented 9.80% of the reasons for consultation in the unit. 73% of the patients were male. The median age was 21. School children and students were the most frequently involved 25.8%) and 58.9% of the patients came from Ouagadougou while 41.1% came from the rural areas. Public thoroughfare accidents (were the main causes of the trauma (25%). Among the traumatic agents, 26% involved vegetal materials. CONCLUSION: Ocular traumas are a major cause of blindness in Western Africa and concern mostly young people. Prevention strategies based on education and information about road safety could reduce the incidence of ocular traumatisms. PMID- 11397981 TI - [Histologic evaluation in rabbits of the biocompatibility of episcleral implant material: S-PTFEe (silicone core covered with expanded polytetrafluoroethylene)]. AB - PURPOSE: To study the tolerance of a silicone band wrapped with expanded polytetrafluoroethylene (e-PTFE) in "fauve de Bourgogne" rabbits. METHODS: A 5.7mmx3.2mm band of S-PTFE (silicone-polytetrafluoroethylene, France Chirurgie Instruments) was used. A tight seal was produced between the two materials by silicone gluing. Eight eyes were implanted episclerally and fixed with Mersilene 5-0. One silicone band was used on a control eye. RESULTS: Three eyes were removed at 4 weeks, one at 8 weeks and four at 12 weeks. No extrusion was observed except for the silicone band rejected 2 weeks after implantation. On histopathologic examination, PTFE colonisation was present in all eyes with fibroblast, inflammatory cells, and neovascularization. CONCLUSION: The S-PTFE implant was colonized and well tolerated in rabbit eyes. Advantages of silicone band (quality, stability of indentation) are combined to the tolerance of PTFE. PMID- 11397982 TI - [Episcleral single injection anesthesia in the caruncle for cataract surgery: report of 350 cases]. AB - Episcleral single injection anesthesia in the caruncle ovoids the rare but potentially dangerous complications and incidents of classic local anesthesia. Excellent motor and sensory block is obtained with a single injection in the anatomic marks are well defined. We have practiced this technique since February 1999 for cataract surgery in 350 cases. The different surgical techniques used were manual extracapsular extraction in 52% of cases, phacosection through tunnel incision in 38%, and phacoemulsification in 10%. Our preliminary results show good acceptance of the episcleral anesthesia for both patients and surgeons. Because it is effective and safe, this anaesthtic technique may be an alternative to classic anesthetic techniques in all cases where topical anesthesia is not indicated. PMID- 11397983 TI - [The Wiedemann-Beckwith syndrome and a congenital cataract]. AB - Wiedemann-Beckwith syndrome is a congenital syndrome with characteristic abnormalities: omphalocele, macroglossia, neonatal gigantism, visceromegaly, hemihypertrophy and a predisposition to embryonic tumors. Ophthalmologic abnormalities have not been described with Wiedemann-Beckwith syndrome. The authors report one case of Wiedemann-Beckwith syndrome associated with bilateral congenital cataract. Family studies indicate linkage of the Wiedemann-Beckwith syndrome locus to the marker 11p15,5. The genetics of cataract is heterogenic. Several mutations responsible for congenital cataract have been described. The association of the Wiedemann-Beckwith syndrome and cataract may contribute to the understanding of the genetics of congenital cataract. PMID- 11397984 TI - [Prognostic outcome of leaking filtering blebs reconstruction with rotational conjunctival flaps]. AB - PURPOSE: Late bleb leaks may follow months to years after filtering surgery especially with the use of antimetabolites. Complications related to beb leaks may lead to a decrease in visual acuity through complicated hypotony or ocular infection. Our retrospective study reports the anatomical and functional results of bleb reconstruction involving the resection of the bleb associated with the covering of the trabeculectomy site with a rotational conjunctival flap. MATERIAL: and methods: Twelve eyes of eleven patients with filtering bleb leaks occurring 3 months to 5 years after successful trabeculectomy (58.3% with adjunct of antimetabolites) underwent bleb surgical reconstruction between november 1995 and June 1999 and were followed until March 2000. Surgical bleb reconstruction was indicated because of persistent or a recurring bleb leak despite conservative medical treatment and blood bleb injections in seven cases. Complications associated with bleb leaks were chronical hypotony (9 cases), athalamy (1 case), hypotony maculopathy (1 case), and endophtalmitis with athalamy (1 case). Three patients had normal IOP but a bleb leak responsible for epiphora. All eyes were treated surgically through bleb excision and conjunctival closure was performed by rotational conjunctival flap. RESULTS: Mean (+/- SD) preoperative IOP was 5.1+/-3.5mmHg (range: 2 to 14mmHg). Mean (+/- SD) postoperative IOP evaluated before any other operation for uncontrolled IOP was 12.7+/-3.1mmHg (range: 6 to 15mmHg). Mean follow-up was 26.7+/-16.9 months (range: 9 to 64 months). All the complications related to the bleb leak resolved after bleb reconstruction. Surgery definively stopped the leak in 10 cases (83.3%) and allowed IOP control without treatment in 50.0% of the cases. Chronic recurring bleb leaks without hypotony occurred in two eyes and required surgery with conjunctival graft which led to a refractory increase in IOP responsible for loss of vision in one case. CONCLUSION: Bleb resection associated with the covering of the trabeculectomy site with a rotational conjunctival flap is a safe and effective procedure for the treatment of a late bleb leak and its complications. In most of the cases (83.3%), long-term IOP control can be expected without, medical treatment in 50% of the cases. Patients must be aware of the possibility of a recurring Seidel; however, the incidence of this complication remains low. PMID- 11397985 TI - [Anterior uveitis and cidofovir]. AB - PURPOSE: This retrospective study was designed to determine the different parameters involved in the occurrence of uveitis during treatment with codofovir. PATIENTS AND METHODS: This study included 10 patients out of 13 treated with cidofovir for cytomegalovirus (CMV) disease. Ocular examination, CD4+ lymphocyte count, and creatinine clearance were performed for each case of uveitis. RESULTS: During the 17-month study, 20 uveitis cases were analyzed. The first attack occurred after a median interval of 7.6 doses. At the time of ocular inflammation, 65% of the cases had a CD4+ lymphocyte count >=100x10(6)/L, the patients thus had an improved immune function. Half of the patients had a normal creatinine clearance. The patients with a CD4+ lymphocyte count<100x10(6)/L who presented one or more incidents of uveitis had an abnormal clearance, thus probably inducing intraocular storage of the drug. CONCLUSION: The occurrence of anterior uveitis during treatment with cidofovir is induced by the association of several parameters: a previous history of CMV retinitis, improvement of the immune function state, and intraocular storage of the drug. PMID- 11397986 TI - [Idiopathic vitreomacular traction syndrome. Vitrectomy results]. AB - PURPOSE: Evaluate the visual results, the complications and the prognosis factors of surgery for idiopathic vitreomacular traction syndrome. METHODS: We retrospectly reviewed 29 consecutive eyes that had undergone pars plana vitrectomy and posterior epiretinal membrane peeling for idiopathic vitreomacular traction syndrome. RESULTS: The mean follow-up after surgery was 17.7 months. Among the 29 cases, 21 were phakic. Visual acuity improved by 2 lignes or more in 72.4% of the cases and the mean improvement was 2.7. Eyes with preoperative visual acuity of 0.25 or more had better postoperative vision than the others (0.42vs0.65;p=0.006). During the follow-up, 10 patients underwent phacoemulsification and posterior chamber implantation (47.6%). The mean visual acuity of these 10 cases was poorer than the mean visual acuity of the 8 pseudophakic patients that underwent vitrectomy (0.45vs0.61;p=0.046). Five recurrences of epiretinal membrane were observed (17.2%). CONCLUSIONS: Surgical treatment of vitreomacular traction syndrome improves visual acuity in most of cases. Further studies with optical coherence tomography are necessary to understand the pathogenic mechanisms of vitreomacular interface syndromes. PMID- 11397987 TI - [Uveal effusion in its pseudotumoral form. A case report]. AB - Uveal effusion exhibits various clinical presentations, some of which may lead to erroneous diagnosis and mimic a choroidal tumor. A 73-year-old female patient displayed a choroidal lesion with a tumoral appearance spontaneously affecting her right eye. Ultrasonography did not establish the diagnosis, leaving doubt for a tumor. In addition to the tumor - like lesion, a moderate inflammation was found in her right eye. The clinical course was favorable and one month after the initial examination, no fundus abnormalities were observed. The delay imposed by the MRI examination contributed to the lack of any active treatment. Apart from cases occurring postoperatively, the diagnosis of uveal effusion may sometimes be difficult to establish. In order to avoid enucleation and even if ultrasonography is helpful in reaching the correct diagnosis, it seems appropriate to wait and regularly follow up the patients when the diagnosis is not certain. The clinical course might be helpful to establish the diagnosis. Our case suggests that in pseudotumoral forms of uveal effusion, we should repeat clinical examination and ultrasonography in order to manage these patients appropriately. PMID- 11397988 TI - [Interferon beta-1b retinopathy during a treatment for multiple sclerosis]. AB - Ocular adverse effects of interferon are described during the treatment of malignant diseases and chronic viral hepatitis with interferon alpha. At this time, there is no report of these effects during multiple sclerosis treatment with interferon beta-1b. The authors report a bilateral retinopathy during this treatment. The production of neutralizing antibodies during interferon beta-1b treatment leads to a decrease in diminution of therapeutic efficacy. When treatment failure occurs, neutralizing antibodies are to be tested. PMID- 11397989 TI - [Gyrate atrophy and craniopharyngioma: a case report]. AB - We report the case of a 13-year-old girl who developed a craniopharyngioma and a gyrate atrophy. No genetic link between these two diseases has ever been reported. This case recalls the characteristic features of gyrate atrophy. PMID- 11397990 TI - [Orbital wooden foreign body]. AB - A 76-year-old-female patient fell in her garden, inducing a right orbital wound in which no foreign body was primarily found. As pain and diplopia followed the orbital trauma, a CT-scan was performed, disclosing a linear external foreign body in the right orbit. An anterior orbitotomy was performed to remove an 18-mm long wooden foreign body. Histopathology of the tissue surrounding the foreign body showed an inflammatory fibrous tissue with small granulomas around telluric particles. Improvement after the operation was dramatic with complete disappearance of diplopia the day following surgery. Antibiotics were prescribed and tetanus prophylaxis was performed. PMID- 11397991 TI - [Bilateral optic neuropathy in acute methanol intoxication. A case report]. AB - We report a case of methanol blindness. Ophthalmoscopic examination disclosed swelling in the disc margins extending along the adjacent retinal nerve fiber layer. Although this optic neuropathy is now rare, prompt diagnosis and proper treatment in the acute phase can dramatically improve the prognosis. PMID- 11397992 TI - [Cyclosporin A eyedrops: manufacturing, toxicity, pharmacokinetics and indications in 2000]. AB - INTRODUCTION: Cyclosporin eye-drops allow local immunoregulation without systemic side effects and is an alternate to local steroids. In this article we review specific problems of product setup and clinical studies published over the past 20 years. PRODUCT SETUP: The main problems in eye-drop preparation are sterility, pH, particles, and its lipophilic properties. Numerous excipients have been tested including oil solvents, alphacyclodextrin, collagen shields, liposomes, polyester nanocapsules, but documentation on stability of the molecule is inadequate. TOXICITY: Epithelial toxicity is well known and is probably mainly due to the excipient. No endothelial toxicity has been described in vivo. Repeated doses lead to uveal reactions in animals, which could limit the indications for intraocular diseases. PHARMACOKINETICS: Bioavailability is mainly limited by the lipophilic properties. Oil excipients, the most widely used, lead to good corneal penetration but low intraocular concentrations. Cyclosporin bioavailability is improved when using hydrophilic excipients. INDICATIONS: Every ocular surface disease that involves cytokines is a potential indication for cyclosporine eyedrops: keratoconjunctivitis sicca, vernal keratitis, adjuvant therapy of filtering surgery, stromal herpes keratitis, immunity limbal keratitis, and Thygeson's keratitis. There is biological evidence of efficacy, and encouraging results from many studies, yet few have tested a large number of patients. A large multicenter study on dry eye is currently in progress. PMID- 11397994 TI - [Esthetics, cosmetics and ophthalmology]. AB - The authors studied many aspects of the cosmetics used around the eye: composition, directions for use, risks for the eyes and their adnexa, and also practical preventive measures. In the future, the cosmetics industry should offer products designed for maximum eye safety. PMID- 11397995 TI - [Human amniotic membrane transplantation in the treatment of ocular surface diseases]. PMID- 11397996 TI - [Hyperkeratosis papilloma of the eyelid. An anatomic clinical case]. AB - A clinicopathologic case of an 80-year-old male patient with a single cutaneous tumor on the upper part of the left eyelid is reported. It was a grayish and pigmented mass covered with a thick keratin layer, well circumscribed, and exophytic. After surgical removal, histopathology showed that the tumor had a papillomatous pattern and was growing under a thick layer of hyperkeratosis. It was a typical squamous cell papilloma. This tumor belongs to the benign eyelid tumor group and can be found on the eyelids of elderly people. PMID- 11397997 TI - [Tocolysis, use of beta-sympatomimetics for threatening preterm delivery: a critical review]. AB - Use of beta-sympatomimetics for threatening preterm delivery: a critical review. Preterm birth is the principal cause of perinatal and neonatal mortality and morbidity. Over the past 40 years, numerous treatments have been tested and used to try to inhibit preterm labor. This literature review is limited to the published studies of beta-adrenergic agonists (beta-sympathomimetics). Among 36 articles that have examined the treatment of acute phase preterm labor by intravenous beta-sympathomimetic administration, 26 are clinical studies with severe biases that make them unacceptable for analysis: non-randomized clinical trials, retrospective studies or retrospective control groups, inadequate recording of pregnancy outcome, exclusion of patients after randomization, confounding factors due to the use of multiple therapeutic agents, inadequate study power.Of the 10 acceptable placebo-controlled trials, seven, including the largest (the Canadian Preterm Labor Investigators' Group, with 708 women), found that beta-sympathomimetics were not better than placebo in prolonging pregnancy or reducing neonatal morbidity. Only three studies found that they were superior to placebo. These agents cause numerous maternal side effects that may be life threatening, because beta-adrenergic receptors are present in many organs. The cardiovascular system is the most severely affected, but side effects also concern the pancreas, kidneys, intestines, and liver. beta-Sympathomimetics cross the placenta rapidly. Fetuses probably respond in the same way their mothers do to stimulation of their beta-adrenergic receptors. Nonetheless, data from the controlled clinical trials show no difference in neonatal mortality or severe morbidity between children born to mothers treated with beta-sympathomimetics and those born to mothers in control groups. The efficacy of preventive treatment by oral or subcutaneous administration of beta-sympathomimetics has also been assessed: a meta-analysis and 2 large randomized placebo-controlled trials have showed that oral maintenance treatment offers no advantages over placebo during the latency phase or for the recurrence rate of preterm labor and the rate of preterm delivery. A single - and smaller - placebo-controlled study found that oral maintenance treatment with ritodrine was beneficial. Treatment trials of subcutaneous administration of beta-sympathomimetics have been performed with a portable subcutaneous pump. Ten studies of this method have been reported, but only two were randomized trials. They found no significant difference in either the mean time until delivery or neonatal morbidity. PMID- 11397998 TI - [Premature labor: tocolysis with non-steroidal anti-inflammatory agents]. AB - Utilization for premature labor of non-steroidal anti-inflammatory agent is well known for thirty years now. Since then, these molecules are regularly tested against others and proposed as efficacious tools. However their prescriptions are not so usual and easy as expected. Very often fetal risks are underlined and exacerbated. For this review of the literature we tried to present on an extensive analysis of complications encountered in clinical practice and ways of avoiding them, following the mechanism of actions of these anti prostaglandins. Perhaps in the near future use of cyclo-oxygenase-type-2-selective non-steroidal anti-inflammatory agent will resolve our dilemma. We present here possible developments. PMID- 11397999 TI - [Tocolysis. Atosiban, an ocytcin-receptor antagonist]. AB - Atosiban, the first member of a new tocolytic drug class, the ocytocin-receptor antagonists, has just been awarded marketing approval in Europe. The clinical effectiveness of atosiban is comparable to that of ritodrine, but atosiban is better tolerated with no evidence of significant maternal or fetal adverse effects. Maintenance therapy with atosiban cannot yet be envisaged due to the lack of a satisfactory formulation at the present time. PMID- 11398000 TI - [Tocolysis with calcium-channel-blockers]. AB - OBJECTIVE: To evaluate the use of calcium-channel-blockers (CCBs) for tocolysis. METHODS: We reviewed the literature retrieved from the Medline database from 1967 to 200 dealing with fetal toxicity and efficacy of CCBs compared with beta adrenergic agonists. RESULTS: Data on fetal toxicity in animals were inconsistent. A teratologic effect has been observed during early pregnancy at supratherapeutic dosage. At usual therapeutic dosage, no fetal abnormalities have been observed. The efficacy of CBs for tocolysis is superior to that of beta adrenergic drugs and allows a reduction of neonatal morbidity. Calcium channel blockers are better tolerated than beta-adrenergic agonists. CONCLUSION: Published data suggest that CCBs could be used a first line tocolytic agents. Although use of CCBs is extremely simple, they should not be prescribed in low risk outpatients. PMID- 11398001 TI - [Tocolysis: dangerous for the infant? Apropos of 1 case and review of the literature]. AB - Tocolysis, commonly used to treat premature labor is mainly achieved with betamimetic agents. The undesirable maternal effects, particularly cardiac effects, of betamimetics are well known but the rare but serious fetal effects are exceptionally reported. We report a case of fetal tachycardia with mild cardiac decompensation and reviewed relevant literature. PMID- 11398002 TI - [Evaluation of clinical competence in gynecology obstetrics: an innovative approach using the Objective Structured Clinical Examination]. AB - OBJECTIVE: The evaluation of medical student clinical competence in Obstetrics and Gynaecology using Objective Structured Clinical Examination (OSCE). MATERIAL AND METHODS: Development of OSCE with definition of an assessment of subject, choice of clinical stations sampling, identification of components of clinical competence to be evaluated, the level of performance required, development of specification table, editing of OSCE presentation page and assessment of praticability and results exploitation. RESULTS: Eleven station stimuli with simulated patient participation were carried out, 2 stations with a mannequin model and 1 questionnaire station. CONCLUSION: Our experience suggests OSCE feasibility for Obstetrics and Gynaecology during the initial course of education presents adequate psychometric characteristics. This gold standard can no longer be overlooked and should be considered as a useful tool to assess medical student competence. Nevertheless, OSCE remains to be further evaluated in France. PMID- 11398003 TI - [Fertility after laparoscopic treatment of ectopic pregnancy in a series of 138 patients]. AB - OBJECTIVE: To evaluate fertility after laparoscopic treatment of ectopic pregnancy (EP). To investigate factors influencing fertility and EP recurrence. PATIENTS AND METHODS: Retrospective study of a series of 138 patients. Rate of intra uterine pregnancy, live births and recurrent EP were the main outcome measures. Subsequent fertility was assessed by calculating cumulative intrauterine pregnancy rates and were analyzed by log-rank tests and Cox regression. RESULTS: The multivariate analysis showed a better rate of intra uterine pregnancy in patients with normal controlateral tube and in women younger than 30. The average time before conception was 11.5 months. After 18 months of infertility the intra-uterine pregnancy rate was low but the EP recurrence rate increased after 24 months. In this study there was no difference between radical and conservative surgery treatment. CONCLUSION: Laparoscopy is useful in the management of ectopic pregnancy, especially in order to establish a prognosis for fertility. PMID- 11398004 TI - [Borderline tumors of the ovary: a multicenter study in 137 patients]. AB - OBJECTIVES: Evaluate management of patients with a borderline ovarian tumor. MATERIALS AND METHODS: A multicentric retrospective survey was conducted in 137 patients with borderline ovarian tumor diagnosed between January 1, 19975 and December 31, 1995. RESULTS: Mean follow-up was 6.5 years, mean age was 50 years. Initial surgery was cystectomy, unilateral salpingo-oophorectomy and total hysterectomy with bilateral salpingo-oophorectomy in 22, 40 and 75 cases respectively. Eleven patients had residual disease. Serous, mucinous and Brenner tumors were observed in 67, 69 and 1 cases respectively. Staging was I, II, III in 117, 3, and 17 cases respectively with two pseudomyxomas. Adjuvant therapy was given in 15 patients. There was a recurrence in 15 patients and 14 died. The 5 year survival rate was 89.3%. Prognosis factors with an impact on survival rate were age, recurrence and type of surgery. Factors with a negative impact on recurrence were adjuvant therapy and residual disease after surgery. CONCLUSION: Careful staging followed by complete and radical surgery is essential. Unilateral salpingo-oophorectomy with omentectomy and multiple peritoneal biopsies may be indicated in younger patients undergoing radical surgery after pregnancy. Aduvant therapy is necessary for invasive implants. PMID- 11398005 TI - [Post-abortum peritonitis pelviperitonitis at the Androva Mahajanga University Hospital: 23 cases]. AB - In many underdeveloped countries, illegal abortion can lead to death or sequelae. Pregnancy termination is illegal in Madagascar except for medical reasons. We report 23 observed between April 1990 and December 1994 at the Androva Mahajanga University Hospital. Post-abortum infectious complications ranked 19th for hospital admissions. Peritonitis was the most frequent (70%) and most serious complication. The women were young (mean age 24 years, 44% under the age of 20 years). Thirty percent were primigravidae and had poor living conditions (74% were homemakers and 13% were students; 44% were single). Clinical signs were malodorous loss and abdominal pain. A surgical procedure was performed in 74% of the cases: 3 salpingo-oophoectomies, 2 hysterectomies and 10 uterine sutures. Mortality was high, 13%. Illegal abortion must be prevented with better information on contraception and better health education programs. The question of the usefulness of current legislation is discussed. PMID- 11398006 TI - [Letter to the editor concerning the article: "Plea for the creation of reference centers for trophoblastic diseases in France" by F. Golfier, L. Frappart, A. M. Schott, D. Raudrant]. PMID- 11398007 TI - [Clinical and pathophysiological contribution of event-related potentials used to study migraine headache]. AB - Event-related potentials are electric brain manifestations evoked by mental activities. This neurophysiological technique is able to describe temporal succession of cognitive processing and allows to measure the neurobiological correlates of each cognitive activity. The evoked potentials of the oddball paradigm and the Contingent Negative Variation (CNV) are also concerned by clinical applications in neuropsychiatry, in neurology and in psychopharmacology. In the case of migraine, the studies with CNV recorded between migraine attacks are characterized by two major phenomena, cerebral hyperreactivity and lack of habituation to repetitive stimuli. From cognitive point of view, this can be interpreted as a difficulty from migraine sufferers to adapt their information processing to environmental constraints. From neurological point of view, this trouble is related with dysregulation of norepinephrin and serotonin ascending pathways. Studies with the oddball paradigm potentials remain non consistent. The mismatch between different methodologies could explain such a lack of consistency. The neurophysiological studies have contributed to new physiopathological hypothesis of migraine. Those hypothesis reveal that a shift in the brain metabolic homeostasis could be the common factor of migraine attacks. The clinical contribution of event-related potentials is of little use in the diagnosis of migraine. But two purposes have been suggested: the differential diagnosis between common migraine and tension-type headaches and the monitoring of beta-blocking agents prophylaxis. PMID- 11398009 TI - [Contribution of positron emission tomography to functional neuroimaging in Alzheimer's disease]. AB - When combined with cognitive investigations, functional neuroimaging methods such as positron emission tomography allow to depict the neural substrates that underlie the neuropsychological alterations in Alzheimer's disease. Capitalising on the variance in both cognitive performances and resting cerebral metabolic rate of glucose (CMRGlc) in Alzheimer's disease, it is possible to correlate these two quantitative variables on a pixel-by-pixel basis and to generate maps showing the significant correlations in stereotaxic space. Some examples using this approach in the domain of memory disorders are presented in this brief review. We notably show that the localisation of the significant correlations differs from one memory system to another, as evaluated by clinical memory tasks. This approach also unravels the compensatory mechanisms that take place with evolution of the disease. Over and above its interest in clinical neuropsychology, this method constitutes a new source of inferences complementary to the classic activation paradigm in normal subjects, as the latter identifies the cerebral structures that are involved with, but not necessarily indispensable for, the normal execution of the task. This approach highlights the interest of combining functional neuroimaging and neuropsychology to better understand the neural substrates of cognitive deficits in both patients with memory disorders and elderly normal subjects. PMID- 11398010 TI - [Dominant forms of Alzheimer's disease: from genotype to phenotype]. AB - This review is devoted to the relationships between phenotype and genotype of the autosomal dominant forms of Alzheimer's disease caused by mutations of presenilins (PSs) and amyloid precursor protein (APP) genes. A first part examines the clinical characteristics mainly the early age of onset and argues about the diversity of age of onset between different mutations and also between patients bearing the same mutation in large families. The second part reports several arguments demonstrating that the main effect of the PSs and APP mutations is the elevation of Ab42 peptide. The pathological and behavioral effects observed in transgenic APP or PSs animals suggest that intra cellular deposits of Ab42 play a role in the pathophysiological process. PMID- 11398011 TI - [Early experience, stressful dysregulation of the hormonal axis and age-related vulnerability to neurodegenerative processus: a longitudinal study in the rat]. AB - The deleterious effects of particular environmental situations have been suspected to augment the repercussions of cerebral injuries leading to increased vulnerability during ageing. The relationship between the hormones of the hypothalamo-pituitary-adrenal axis, mainly glucocorticosteroids, and cerebral structures like the hippocampus has been the subject of intense investigation in the recent years. Data suggest that long-term elevated blood levels of these hormones can induce neuronal alterations leading to cognitive dysfunction. This hypothesis has been tested with relevant animal models of normal/abnormal ageing. The models are based on the existence of considerable inter-individual differences in the degree of age-related cognitive impairments observed in rodents. Results show that long-term glucocorticosteroid exposure induces cerebral changes related to the action of these hormones on their central receptors. Experimental data are in accordance with clinical investigations suggesting that hormonal changes, and chronic life events, could be considered as a predictive factor of future cognitive dysfunction. PMID- 11398012 TI - [Memory and executive functions in multiple sclerosis: preliminary findings with a cognitive battery]. AB - Twenty one MS patients suffering from a relapsing-remitting (four patients), a secondary progressive (12 patients) or a primary progressive form of the disease (five patients) were assessed using a cognitive battery specifically devoted to executive and memory processes. Results showed that, patients without significant depressive state evaluated by the MADR scale, exhibited significant impairments in executive processes, working, episodic and procedural memories whereas short term memory, language and global intellectual efficiency were normal. The preliminary data suggest an impairment of encoding, a process previously underestimated in this disease. In addition, the battery was easily administered to the patients and relevant for the cognitive assessment of MS. PMID- 11398013 TI - [Taste impairment in Alzheimer's disease]. AB - We conducted a study of Alzheimer's disease focusing on taste impairment to determine at which step of gustatory information processing (discriminative or perceptive, gnosic or associative, gustatory-verbal) possible perturbations might be located. We used various foods found in a normal diet. Twenty patients and a matched control group participated in this study. The patients were divided into two subgroups according to disease severity (mild, moderate or severe). The study showed that patients with Alzheimer's disease exhibited an impairment at all three levels of gustatory information processing. Impairment of discriminative and gustatory-verbal stages was more obvious in the group of patients with severe Alzheimer's disease. Our study also showed that the gustatory deficiency can be described as an associative agnosia in the mild stage of Alzheimer's disease. This concept is in line with the notion of a dissociation between preservation of olfactory and gustatory thresholds and an alteration in odor identification in patients with mild stage Alzheimer's disease, suggesting that the alteration is central rather than peripheral. Our findings suggest that an alteration of the associative level of gustatory information processing can be found in Alzheimer's disease early stage. PMID- 11398014 TI - [Unilateral spatial and motor neglect in children]. AB - Although unilateral spatial neglect is well documented in adults after a cerebral damage, the description in children is rare and no motor neglect has yet been reported. We report four children age range from three to thirteen years old, in patients in a rehabilitation department. A cancellation task "the Teddy bear test" was used to assess spatial neglect in children. The children were presented a page containing line drawings of usual objects, including fifteen Teddy bears. The children were asked to cross out each Teddy bear. These four children omitted five to eleven teddy bears located on the left space, whereas children of the controlled group never omitted more than one Teddy bear. The four children also exhibited neglect in drawings, and during daily life activities. Moreover, one of them had a motor neglect with no more defects of strength and an under utilization of his left side. Spatial unilateral neglect and motor neglect should be systematically investigated in children after a cerebral damage, and should be considered during rehabilitation. PMID- 11398015 TI - [Arterial thrombolysis and angioplasty in atheromatous basilar artery occlusion]. AB - Basilar occlusions treated with conventional therapy (anticoagulants or antiplatelets) have a poor outcome (80p. cent mortality). This unfavorable outcome may require a treatment within 6 hours by intra-arterial thrombolysis, sometimes followed by percutaneous transluminal angioplasty (PTA) in case of atherothrombotic occlusion due to a tight atherosclerotic stenosis. A 48 year-old patient, presented with left hemiparesis, left multimodal hypoesthesia, paralytic dysarthria. CT-scan showed a spontaneous hyperdensity of the basilar artery and arterial occclusion was confirmed by angiography, which showed an atherothrombotic occlusion involving the proximal part of the vessel. Intra arterial thrombolysis began five hours after the onset with 0,25mg/kg of Rt-Pa (Actilyse((R))), given by bolus followed by 4 others bolus of 10mg. After a total dose of 60mg, arterial recanalization was obtained showing a tight atherosclerotic stenosis involving the proximal part of the basilar artery. PTA was performed 18 hours later with a ballon inflation at 6 atmospheres during 20 seconds. It allowed to decrease the stenosis from 80p. cent to 60p. cent. The patient recovered and MRI at D20 showed a small right lateral infarct involving the pons. Our study confirms the usefulness of intra-arterial thrombolysis in basilar artery occlusion. Consecutive PTA may be proposed in case of associated atherosclerotic stenosis, and the interest of PTA is further discussed. PMID- 11398016 TI - [A case of mitochondrial cytopathy (MERFF) without ragged red fibers at the onset]. AB - We report a case of a mitochondrial cytopathy (MERRF) with a late diagnosis because of the absence of Ragged-Red Fibers (RRF) in the first muscular biopsy performed in childhood. Eight years after the onset of the disease the familial history and the clinical manifestations were very suggestive of a mitochondrial cytopathy and a second biopsy was performed confirming the diagnosis of MERRF. The authors discuss several hypotheses to explain the negativity of the first muscular biopsy. The absence of ragged-red fibers does not exclude the diagnosis of a mitochondrial cytopathy and other investigations are justified such as the biochemical study of the respiratory chain and the molecular analysis of mitochondrial DNA. PMID- 11398017 TI - [Idiopathic orbital myositis]. AB - Orbital myositis is a rare disorder considered as a subgroup of inflammatory orbital pseudotumors. The pathophysiology is still unknown. Patients typically present with orbital pain exacerbated by eye movement and diplopia. Response to steroids is dramatic. We report a case of idiopathic myositis of the right inferior muscle, which recovered after steroid therapy. PMID- 11398018 TI - [Cerebral cysticercosis and headache in a young Togolese woman]. PMID- 11398019 TI - [History of dystonia]. PMID- 11398020 TI - [Psychological impact of oligodendroglioma]. PMID- 11398021 TI - [Generalities on the molecular mechanisms of apoptosis]. PMID- 11398022 TI - Transoesophageal echocardiography for minimally invasive cardiac surgery. PMID- 11398023 TI - Scanning electron microscopic analysis of endoscopic versus open vein harvesting techniques. AB - BACKGROUND: Endoscopic vein harvesting techniques are increasingly used for obtaining conduit for coronary artery bypass surgery. Although they offer advantages in healing over the conventional open technique, concern has been raised regarding the potential for trauma to the vein in the form of intimal disruption which would theoretically predispose to early graft thrombosis and/or development of stenoses. Unfortunately no long term data is yet available for determining if conduits harvested in this fashion are prone to such events. METHODS: We have examined vein segments harvested by both endoscopic and open techniques for evidence of intimal injury (either visible disruption of the intima and/or presence of thrombus) using scanning electron microscopy (SEM). Those harvesting the vein were unaware which patients were in the study, and both the SEM technician and cardiac pathologist who evaluated the scans were blinded to the technique used for harvesting. For each vein segment examined, views were obtained of four different sections and were analyzed at magnifications ranging from 10 yen to 100 yen. RESULTS: Both thrombus formation and visible intimal disruption were identified quite rarely, and overall were not linked significantly to the type of harvesting technique used. CONCLUSIONS: These results suggest that endoscopic vein harvesting techniques do not subject the conduits to more trauma than open techniques and therefore may not predispose to the development of earlier stenoses. This data will need to be confirmed by both other methods of identifying intimal injury and by long-term follow-up of conduit patency in both groups. PMID- 11398024 TI - Doppler echocardiographic evaluation of St. Jude Medical valves in the tricuspid position: criteria for normal and abnormal valve function. AB - BACKGROUND: Although this examination for mitral and aortic prosthetic valves has been frequently reported to be available for the early diagnosis of complications, there are few reports on examinations for tricuspid prosthetic valves, particularly the mechanical valve such as the St. Jude Medical (SJM) valve. Furthermore, the criteria for early diagnosis of complications are unknown. The purpose of this study is to evaluate the SJM valve in the tricuspid position by Doppler echocardiography and to analyze valve-related complications. METHODS: Between September 1978 and March 1997, 46 patients underwent tricuspid valve replacements with SJM valves; these cases were evaluated by Doppler echocardiography. We divided the cases into Normal and Complication groups. RESULTS: In the Normal group, peak pressure gradient was 4.2+/-1.5 mmHg and pressure half-time was 123.5+/-22.9 msec. In the Complication group, peak pressure gradient was 12.7+/-3.6 mmHg and pressure half-time was 271.8+/-76.4 msec. In the Complication group, the lowest peak pressure gradient case was 8.0 mmHg and the shortest pressure half-time was 160.0 msec. CONCLUSIONS: We concluded that when the peak pressure gradient is more than 8 mmHg or the pressure half-time is more than 160 msec, there is a possibility of complications. Despite the problems, the absence of any structural change after more than ten years suggests that the SJM valve can be used in the tricuspid position with careful follow-up. PMID- 11398025 TI - Conventional coronary artery bypass grafting: why women take longer to recover. AB - BACKGROUND: Recovery following successful coronary artery bypass grafting (CABG) has been dramatically improved with the use of fast-track methods. Although data exist that demonstrate a significant gender difference in survival following CABG, little is known about factors influencing gender-specific recovery. This report describes a series of consecutive patients undergoing isolated CABG to determine gender-associated factors that may impact outcomes and recovery. METHODS: Five hundred and seventeen consecutive patients underwent isolated CABG utilizing cardiopulmonary bypass and were retrospectively reviewed. The outcomes of 351 men in the study were compared to the group of 160 women. A rapid recovery protocol focused on reduced cardiopulmonary bypass time, aggressive preoperative intra-aortic balloon pump use, early extubation, perioperative administration of corticosteroids and thyroid hormone, aggressive diuresis and atrial fibrillation prevention was applied to all patients. RESULTS: The 30-day mortality rate for the women was 4.2% (Parsonnet risk 16.3+/-9.0) compared with 3.4% (Parsonnet risk 9.9+/-7.5) for the men. There were no statistically significant differences in the 30-day mortality rates or postoperative complication rates between the women and men. The women, however, were found to be older (71+/- years versus 65+/- years, p<0.001), and to have a higher incidence of acute myocardial infarction (31% versus 20%, p<0.05), obesity (23% versus 10%, p <0.05), diabetes (31% versus 22%, p<0.05), hypertension (65% versus 48%, p<0.001), and symptomatic vascular disease (20% versus 12%, p<0.05). The women required fewer bypass grafts (2.9 versus 3.5 grafts, p<0.001), and consequently, had shorter cross and cardiopulmonary bypass times. Rapid recovery with discharge before the fifth postoperative day was achieved in 30% of the women, in comparison to 44% of the men (p<0.01). The postoperative hospital length of stay was longer for the women in comparison to the men (7.2+/-7.1 versus 5.8+/-5.2 days, p<0.05). CONCLUSIONS: Women had similar operative mortality and postoperative complication rates to men under a rapid recovery protocol. However, women have a longer recovery interval compared to men, which may be a reflection of their higher preoperative risk profile. PMID- 11398026 TI - Aortic valve replacement with the composite Labcor porcine bioprosthesis in the elderly. AB - BACKGROUND: This paper presents the analysis of clinical results of the composite porcine Labcor bioprosthesis in the replacement of aortic valves in the elderly. METHODS: This retrospective study was carried out in the Thoracic and Cardiovascular Surgical Department, La Pitie-Salpetriere Hospital, Paris, for replacement of calcified, stenosed aortic valves between 1988 and 1995. It involved a series of 100 patients aged 70 and over (mean: 80+/-5 years ranging from 70 to 90). There were 63 female and 37 male patients. Preoperatively, five patients were in NYHA Class I, 23 in Class II, 65 in Class III and 7 in Class IV. RESULTS: Fifteen patients died in the early postoperative stage and 13 during the follow-up period. There was no evidence of valve failure. The average follow-up was 32 months and the actuarial survival rate at 5 years was 74+/-5%. Complications due to bleeding occurred in 3 patients taking anticoagulant treatment. There were neither valvular thrombosis nor embolism. Two patients presented with prosthetic endocarditis. Two patients received a reoperation because of leakage (1 septic). The five-year follow-up showed that 96% of patients did not require further surgery. When this study was completed, 83% of patients were in Class I or II versus 71% in Class III or IV prior to surgery. CONCLUSIONS: In the early/middle follow-up term, the results obtained when replacing the aortic valve with the composite Labcor bioprosthesis in the elderly are satisfying. Nevertheless, further long-term assessment is needed. PMID- 11398027 TI - Differences in prolonged ischemia length using ischemic preconditioning in the rabbit heart. Tolerable limitation time for surgically induced myocardial ischemia during normothermic cardiac operation. AB - BACKGROUND: To determine the effect of the tolerable limitation time of prolonged ischemia after ischemic preconditioning on postischemic functional recovery and infarct size reduction in the rabbit heart. METHODS: White rabbits (n=30) were used for Langendorff perfusion. Control hearts were perfused at 37 degrees C for 180 min; 30 min global ischemia hearts (30GI) received 30 min global ischemia and 120 min reperfusion; IPC+30GI hearts received 5 min zero flow global ischemia and 5 min reperfusion prior to 30 min global ischemia; 20 min global ischemia hearts (20GI) received 20 min global ischemia and 120 min reperfusion; IPC+20GI hearts received 5 min zero flow global ischemia and 5 min reperfusion prior to 20 min global ischemia. RESULTS: Infarct size in the 30GI hearts was 33.5+/-4.0% and 1.7+/-0.5% in the control hearts. The 20GI hearts and IPC+30GI hearts decreased infarct size, as compared with the 30GI hearts (13.0+/-1.8% and 16.6+/-1.7%, respectively; p<0.001, 20GI vs 30GI; p<0.01, IPC+30GI vs 30GI; p>0.05, 20GI vs IPC+30GI) but did not enhance postischemic functional recovery. The IPC+20GI hearts (3.5+/-0.6%) significantly decreased infarct size as compared with the 20GI hearts (p<0.05, IPC+20GI vs 20GI), and there was no significant difference between the IPC+20GI and the control hearts (p>0.05), but the IPC+20GI hearts did not enhance postischemic functional recovery. CONCLUSIONS: A 20 min ischemia may be the tolerable limitation time of prolonged ischemia after ischemic preconditioning in an isolated rabbit heart model. PMID- 11398028 TI - Postoperative myocardial ischemia in thoracic aortic aneurysms. AB - BACKGROUND: To determine the incidence and predictors of postoperative myocardial ischemia in non-coronary risk patients undergoing surgery for thoracic aortic aneurysms. METHODS: DESIGN: a prospective, observational study. SETTING: a general intensive care unit in a university hospital. PARTICIPANTS: twenty patients without ischemic heart disease, scheduled for elective surgical repair of thoracic or thoracoabdominal aortic aneurysms. INTERVENTIONS: all patients underwent aortic replacement with prosthetic graft and routine postoperative care. Patients who developed myocardial ischemia received an infusion of coronary vasodilators. RESULTS: ECG episodes of myocardial ischemia were defined as reversible ST-segment changes of either >1 mm of depression or >2 mm of elevation at the J point. All patients survived operation. Eleven patients (ischemia group) developed myocardial ischemia, and 9 patients did not (non-ischemia group). These episodes were transient in 8 cases, but lasted longer than 3 days in 3 cases. In univariate analysis of perioperative factors between the two groups, the use of total cardiopulmonary bypass (p<0.01), the cardiac index at ICU admission (p<0.05), and the incidence of pre-existent hypertension (p<0.05) were significantly different. Multiple regression analysis identified the use of total cardiopulmonary bypass as the only predictor of myocardial ischemia. CONCLUSIONS: The use of total cardiopulmonary bypass is predictive of perioperative myocardial ischemia in surgery for thoracic aortic aneurysms, probably due to the production of proinflammatory cytokines by systemic ischemia and reperfusion. Prophylactic use of coronary vasodilators may be validated in these cases. PMID- 11398029 TI - Predictive value of Candida colonization index in 131 patients undergoing two different cardiovascular surgical procedures. AB - BACKGROUND: Nosocomial candidemia is an important infection because of its increasing incidence and its high fatality rate. Candidiasis involves multiple risk factors. In this work, we study the degree of Candida colonization in cardiovascular surgical patients by taking into account the number of sites colonized and the density of growth. METHODS: Eleven (11) selected variables (age, sex, weight, diabetes, number of antibiotics, duration of antibiotics, length of hospitalization, length of stay in surgical intensive care unit, duration of surgery, temperature during surgery, and number of bypass) were considered to predict the perioperative variation of the colonization index (CI) by Candida in 131 cardiovascular surgical patients. These patients were divided into two groups: group A, coronary artery bypass grafting with extracorporeal circulation (72 patients) and group B, coronary artery bypass grafting without extracorporeal circulation (59 patients). RESULTS: One thousand and forty-eight fungal cultures were obtained from four different body sites and 162 isolated were identified. Candida albicans accounted for 74% of the strains in group A and 97% in group B. The statistical analysis (two-way anova) shows that group A patients with an increased CI have received significantly more antibiotics than those with a stable CI (1.50+/-0.83 vs 1.08+/- 0.40, p=0.0055). CONCLUSIONS: Epidemiological data obtained from this study show that coronary artery bypass grafting with extracorporeal circulation procedure is associated with an increase in the use of antibiotics and subsequently a higher risk a Candida colonization infection. PMID- 11398030 TI - Recurrent cardiac myxoma: why it occurs. A case report with literature review. AB - Two years after surgical removal of a right atrial myxoma, recurrences were found in both atria: multiple tumors in the right atrium and one tumor in the left atrium. All were subsequently removed. There was no further recurrence during the following 3 years. We report on an unusual case which, to our knowledge, is the first time that such a case appears in the literature. We also discuss the clinical symptoms, diagnostic aspects, as well as details of a literature review regarding recurring myxoma. We think the recurrence in our case was due to totipotent multicentricity of the tumor. Previous studies showed that the recurrence might be due to the following reasons: inadequate resection, or multifocal pattern behaviour of a benign myxoma, either in the same or in a different location as the primary tumor. Familial disposition may also play a role in recurrent development. The abnormal DNA ploidy pattern of myxoma patients showed a high recurrence. PMID- 11398031 TI - Coronary artery bypass grafting in a patient with brainstem ischemia. AB - We describe a 54-year-old male with severe coronary artery disease and cerebrovascular disease including right cerebellar infarction, total occlusion of the bilateral vertebral arteries, brainstem ischemia, and right cerebral infarction with significant right carotid artery disease. Repeated percutaneous transluminal coronary angioplasty had been performed, however, unstable angina was developed despite maximal medical treatment. Coronary artery bypass grafting was successfully undergone with use of propofol, application of the intra-aortic balloon pumping perioperatively, and mild hypothermic cardiopulmonary bypass with alpha-stat blood gas management. The importance of preoperative evaluation of the intracranial circulation and management of cardiopulmonary bypass are discussed. PMID- 11398033 TI - Reconstruction of the median sternotomy wound dehiscence using the latissimus dorsi myocutaneous flap. AB - BACKGROUND: Currently the internal thoracic artery (ITA) is the conduit of preference for coronary artery revascularization. Although this artery offers several advantages over the saphenous vein there is a higher incidence of postoperative sternal wound infection with its use. This incidence further increases with the use of bilateral internal thoracic arteries (BITA). The use of muscle or omental flaps to treat this complication has significantly reduced the morbidity and mortality. Typically the pectoralis major (PM) or the rectus abdominis (RA) muscles are the preferred method of reconstruction of the infected sternotomy wound. METHODS: In a retrospective study over a four-year period from February 1994 to October 1998, nine patients underwent reconstruction of an infected median sternotomy wound with a latissimus dorsi myocutaneous flap (LDMF). RESULTS: All of the patients in our study were successfully treated with a single LDMF with the exception of one who required a rectus abdominis flap to cover the lateral aspect of the recalcitrant poststernotomy infected wound. There was a single patient who had a wound dehiscence at the donor site. CONCLUSIONS: The LDMF is reliable and serves as an adjunct for treating sternotomy infections. The flap provides sufficient amount of pedicle length and muscle mass for coverage. Although there is a need to turn the patient into a lateral decubitus position once the debridement is performed the flap harvest and its mobilization is technically straightforward with a short operative time, 135 minutes on average with a range of 97 to 171 minutes. Furthermore, there exists an anatomical advantage in using the LDMF; harvest of the LDMF does not disrupt collateral blood supply to the sternum and parasternal tissues. PMID- 11398032 TI - Technique of transmyocardial revascularization: avoiding complications in high risk patients. AB - BACKGROUND: Transmyocardial revascularization (TMR) is an emerging therapy for coronary artery disease, with 7 years of published clinical research and nearly three years of clinical application. Every report thus far has confirmed that TMR relieves severe angina pectoris. It is primarily an alternative therapy for angina which has been refractory to conventional medical and surgical treatment. Operative mortality of 3% to 10% has been reported. METHODS: Seventy-seven patients were treated with TMR using a Holmium: YAG laser. Admission criteria included severe angina despite high doses of at least two anti-angina medications and nitroglycerin, reversible ischemia by thallium scan, and unsuitability for CABG or angioplasty. Patients had end-stage ischemic heart disease and multiple previous conventional procedures. TMR was performed through small left anterior thoracotomies using a 10.16 cm or less incision. RESULTS: Seventy-five patients recovered from surgery without major complications. One patient (1.3%) died of an autopsy-proven myocardial infarction in the treated region, and two additional patients had a myocardial infarction (4.3%). Four patients had paresis of the left phrenic nerve, as determined by an elevated left hemi-diaphragm on chest X ray, from which three recovered fully. Patients had no bleeding or wound infections. Patients were intubated for an average of 1.5 hours and remained in ICU an average 0.8 days. Mean hospitalization was 3.4 days. CONCLUSIONS: Technically well-done TMR through a small anterior thoracotomy can have good therapeutic results and low morbidity and mortality. We will describe operative techniques which minimize pain and stress and help to insure surgical success. PMID- 11398034 TI - Management of carotid coils during routine carotid endarterectomy. AB - BACKGROUND: A coil in the internal carotid artery (ICA), defined as a circular configuration or exaggerated S shape of the ICA, is occasionally encountered during endarterectomy for carotid bifurcation lesions. The significance of coils as an etiology for symptoms is difficult to determine. It is thought, however, that the failure to correct coils and kinks during routine carotid endarterectomy (CE) may lead to turbulence and failure of the CE. Various techniques have been discussed to repair coils. METHODS: Our technique consisted of complete dissection of the coil, routine use of a Javid shunt, standard endarterectomy, resection of the redundant ICA, re-approximation of the posterior wall of the ICA and patch angioplasty of the anterior wall. Three hundred and fifteen patients underwent CE between August, 1998 and February, 2000. Fifteen patients (4.7%) had a carotid coil that was repaired. There were ten men and five women. Mean age was 72.6+/-6.1 years. Ten patients had an asymptomatic stenosis. Four patients had lateralizing symptoms and one patient had dizziness. Fifteen patients underwent preoperative duplex scanning and 14 of these patients had MRA scans performed. All patients had a preoperative stenosis of 80-99% by duplex on the operated side. The right carotid artery was repaired in 12 patients. The left in three patients. The length of resected artery varied from 1.2-2.8 cm (1.93+/-0.49 cm). RESULTS: All patients survived surgery. One patient developed a cerebellar stroke on the third postoperative day. A postoperative carotid duplex scan demonstrated a widely patent repair. There were no cranial nerve injuries in this series. One patient died seven months after surgery from cardiac events with no follow-up duplex exam. There have been no long term strokes or anastomotic complications. Follow-up duplex scans demonstrated widely patent repairs (1-15% stenosis) in seven patients and low end 15-49% stenosis in five patients. CONCLUSIONS: Resection of redundant ICA with re-anastomosis of the posterior wall and patch reconstruction of the anterior wall gives acceptable perioperative and long term results. PMID- 11398035 TI - Value of stent placement during percutaneous transluminal angioplasty of the iliac arteries. AB - BACKGROUND: To determine the benefits of stents during percutaneous transluminal angioplasty (PTA) of the iliac arteries. Retrospective analysis of our 10-year surgical experience with iliac PTA from 1988-1997 permitted comparison of results during two consecutive periods: an initial period (1988-1992), during which stents were never used, followed by a second period (1992-1997), when stenting was performed on indication. METHODS: From January 1988 to October 1997, 287 iliac PTA procedures (158 common iliac arteries, 129 external iliac arteries) were performed on 250 patients. Thirty-seven patients had two iliac lesions that were treated simultaneously. Indications for PTA included stenosis (270 cases) and chronic occlusion (17 cases). Thirty-one patients (12.4%) underwent infra inguinal bypass in addition to PTA owing to the existence of arterial lesions at two levels. Two consecutive patient groups were defined: Group I consisted of 75 patients who underwent 86 iliac PTA procedures between January 1988 and May 1992 without stent placement; Group II consisted of 175 patients who underwent 201 iliac PTA procedures between June 1992 and October 1997, when selective stenting was performed. A total of 55 stents (35 in the common iliac artery, 20 in the external iliac artery) were placed in Group II during PTA due to unsatisfactory immediate results (dissection, residual stenosis) or occlusion. RESULTS: There was no 30-day mortality. There were 15 immediate failures of PTA: 8 in Group I (10.7%) and 7 in Group II (4%). The cumulative initial success rate was 94%. Follow-up ranged from 3 months to 102 months (mean 37 months). The cumulative primary patency rate at 4 years was 62% (58% in Group I, 64% in Group II). The cumulative secondary patency rate at 4 years (including patients who subsequently underwent repeat angioplasty procedures) was 72% (68% in Group I, 74% in Group II). CONCLUSIONS: Stents were an effective means for treatment of initial failures of PTA in patients with iliac artery occlusive disease. However, there were no significant differences in the long term results between PTA alone and PTA with selective stent placement. PMID- 11398036 TI - Is arterial surgery advisable for patients over 80 years of age? AB - BACKGROUND: Recently life expectancy has become longer and longer. The purpose of this study was to analyse whether arterial surgery for patients over 80 years of age is advisable. METHODS: During the last 14 years, 527 patients, 50 of whom were over 80 and 477 of whom were under 80 years of age, received graft replacement or bypass surgery. They suffered from ruptured abdominal aortic aneurysm (R-AAA, n=21), non-ruptured abdominal aortic aneurysm (N-R AAA, n=133) or arteriosclerosis obliterans (ASO, n=373). Complications such as cerebrovascular disease, ischemic heart disease, respiratory and kidney dysfunction, and risk factors for ASO were also checked. RESULTS: All of the patients over 80 with R-AAA (n=3/3) and 50% of the patients under 80 with R-AAA (n=9/18) died during their stay in the hospital. However, none of the N-R AAA patients over 80 (n=0/7) and only one of the 126 N-R AAA patients (0.8%) under 80 died. For the patients over 80 with ASO, the graft patency rate was better than the patients survival rate. There were no age-specific factors that should condemn arterial surgery for patients over 80 years of age. CONCLUSIONS: Arterial surgery should not be ruled out on the basis of age alone. PMID- 11398037 TI - Comparison of femorofemoral and aortofemoral bypass for aortoiliac occlusive disease. AB - BACKGROUND: Role and results of femorofemoral bypass grafting, usually reserved to high-risk patients affected with unilateral iliac artery occlusion, are still debated. METHODS: EXPERIMENTAL DESIGN: retrospective clinical study. SETTINGS: University Hospital. PATIENTS: seventy-six high-risk patients (group 1) who underwent a primary expanded polytetrafluoroethylene (ePTFE) externally supported femorofemoral bypass graft were retrospectively compared to two additional groups of patients selected from the entire series of patients who underwent an aortobifemoral bypass graft. Patients of group 2 (n=80) were randomly chosen to determine differences in risk factors, associated diseases, previous abdominal operations, operative indications, preoperative findings and outcome. Patients of group 3 (n=50) were matched for sex, risk factors, associated diseases, previous abdominal operations, operative indications and preoperative findings with those of group 1 to assess the importance of the type of operation in determining the outcome of the procedure. RESULTS: Postoperative mortality (6, 4 and 6%, respectively), 5-year primary and secondary patency (71, 80, 83% and 80, 87, 87%, respectively) and limb salvage rates (78, 87 and 87%, respectively) were similar among the groups (p=NS, p=NS, p=NS, respectively). Five-year survival rate of group 2 was significantly better than that of group 1 and 3 (p<0.04 and p<0.04, respectively). CONCLUSIONS: Primary ePTFE externally supported femorofemoral bypass graft in high-risk patients is safe and produces long-term results similar to aortofemoral reconstruction. PMID- 11398038 TI - Continuing expansion of internal iliac artery aneurysms after surgical exclusion of the inflow. A report of two cases. AB - Two patients with continuing expansion of an internal iliac artery aneurysm following earlier repair of an aortoiliac aneurysm are described. At the primary operation, inflow to small internal iliac aneurysms had been interrupted by simple proximal ligation only. During follow-up, however, increasing diameter of the by CT-angiography completely thrombosed internal iliac aneurysms required re operation. The observation of continuing growth of thrombosed internal iliac artery aneurysms following proximal ligation emphasises the danger of persistent collateral circulation and supports the concept of endotension in the absence of endoleak following endovascular AAA repair. PMID- 11398039 TI - Combined percutaneous and surgical approach to a postnephrectomy arteriovenous fistula. AB - The case of a patient with long-standing severe heart failure that was ultimately found to be secondary to a large communication between the right renal artery stump and the inferior vena cava is reported. The occurrence of an arteriovenous fistula should be suspected in patients who underwent previous nephrectomy since early diagnosis would avoid relevant cardiac complications and the demise of a patient suffering from a disease that may be corrected surgically or percutaneously. We describe a combined approach that allowed the percutaneous decompression of the inferior vena cava and relief of the volume overload to the heart prior to the surgical ligation of the arteriovenous fistula. This approach should be considered a valuable option for treating long-standing high output postnephrectomy arteriovenous fistulas. PMID- 11398040 TI - Replacement of the entire thoracic aorta according to the reversed Elephant Trunk technique. AB - The aim of this work is to present our modified Elephant Trunk technique to reduce circulatory arrest time and consequently mortality and morbidity rates. According to Borst's technique the ascending aorta and aortic arch are replaced first, under deep hypothermic circulatory arrest, while a graft segment is left in the descending thoracic aorta. In the second stage of the operation, the descending thoracic aorta is replaced through left thoracotomy using this graft segment. In our modified technique, after the flexion in the proximal segment of the graft, the descending thoracic aorta is replaced first through left thoracotomy in Bio-Pump protection, choosing the best aortic segment for proximal anastomosis. In the second stage we replace the ascending aorta and the aortic arch using the graft and applying Carrel patch anastamosis only to the epiaortic vessels, under deep hypothermic circulatory arrest. It is our opinion that the mortality incidence of this technique is similar to that obtained with Borst's technique, though certainly inferior to the one stage procedure , while the morbidity results are better than those obtained with the Borst Elephant Trunk technique and with the one stage procedure. In fact there are fewer stroke incidents thanks to the reduced times of deep hypothermic circulatory arrest, and fewer postoperative bleedings and respiratory failures thanks to the reduced times of the total cardiopulmonary bypass. At the beginning we used this technique to replace symptomatic aneurysms, covered ruptures, and hematomas of the wall of the descending thoracic aorta, which required replacement of the descending thoracic aorta first; we later extended the treatment to all types of thoracic aorta aneurysms. PMID- 11398041 TI - Early experience with PET scanning in thoracic tumours. AB - BACKGROUND: Positron emission tomography (PET) scan is an imaging technique which relies on metabolic properties of the lesions. In this study, we evaluated the potential contribution of PET for thoracic malignancy in a consecutive series of patients presenting with multiple lesions or equivocal diagnosis. METHODS: PET with 2-18 F fluorodeoxyglucose (FDG) was carried out in 41 patients. The diagnosis was primary intrathoracic malignancy in 22 (Group 1). On routine staging using CT scan we found 29 additional lesions and assessed these using PET scan. PET was performed to evaluate the number of metastatic lesions in the lung in 11 (Group 2), to characterise undiagnosed pathology in the chest in 4 (Group 3), to search clinically suspicious extrathoracic spread in 4 patients with known intrathoracic malignancy. RESULTS: In Group 1, the sensitivity and specificity of PET was 81.2% and 92.3%. The accuracy of PET in the confirmation of metastatic disease to the chest was 73%. PET was falsely positive in a patient with chronic inflammatory disease in Group 3 and highly accurate to characterise unknown pathology in Group 4. CONCLUSIONS: Even though infection may cause false positive results, PET is a useful imaging technique for the evaluation of patients with thoracic tumours. PMID- 11398042 TI - Surgical treatment of primary and metastatic sternal tumours. AB - BACKGROUND: Primary and metastatic malignancies of the sternum are uncommon. Surgery that is the best treatment for the majority of primary sternal tumors, and arguably for metastatic lesions, has improved permitting us to perform wide resection and simultaneous reconstruction safely. METHODS: From January 1988 to December 1998 we treated 13 patients, 4 with primary chondrosarcoma and 9 with sternal metastasis, 5 breast cancer, 3 kidney cancer and 1 thyroid cancer. In 3 patients total sternectomy was performed and in 10 a partial sternectomy associated with resection of the anterior segment of the ribs in 7 cases and resection of the clavicle in 5 patients. Bone reconstruction was done in the majority of cases (5) with Marlex mesh with methylmethacrylate and in 3 cases rib grafts were used to strengthen a Vicryl mesh. The major pectoralis muscle was the most frequently used soft tissue, 9 of 12. RESULTS: Our postoperative mortality was 15%, 2 cases. The median overall survival was 48 months. All the primary tumours were alive after a mean follow-up of 34 months (range 4-84 months). While survival of the sternal metastasis was 24 months. CONCLUSIONS: Surgical resection and reconstruction of sternal lesions represent a basic step in the treatment of the primary tumors with encouraging survival results while in the metastatic lesions surgery can be a part of a multimodality approach with unsatisfactory results. PMID- 11398043 TI - Pericardial catheter sclerosis versus surgical procedures for pericardial effusions in cancer patients. AB - BACKGROUND: Approximately 21% of patients with advanced malignancies have cardiac or pericardial involvement with tumor. Controversy exists regarding the optimal approach to the pericardial space when hemodynamic compromise due to effusions occurs. METHODS: A six-year retrospective review of 59 cancer patients with pericardial effusions. RESULTS: Thirty-six patients had subxiphoid pericardial window (SXPW) alone (Group A), 5 had pericardial catheter drainage (PCD) followed by a SXPW (Group B), 10 had PCD with sclerosis (Group C), 5 had PCD alone (Group D), 2 had PCD with pericardial-pleural window (Group E), and one had pericardial peritoneal window (Group F). The method of procedure, complications, number of hospital and ICU days, cytological or pathologic evidence of malignancy, solid versus hematological tumors, and survival were analyzed. The median survival for those patients in group C was one month compared to 4 months for Group A and 6 months for Group B. Essentially, results were similar regardless of method performed with the exception that professional and hospital charges averaged $4830 for SXPW compared to $1625 for PCD. CONCLUSIONS: Pericardial catheter drainage and sclerosis provides a viable option for the treatment of pericardial effusions in selected cancer patients at markedly reduced cost and patient discomfort. PMID- 11398044 TI - The bilobectomies for lung cancer. AB - BACKGROUND: The purpose of our retrospective study is to confirm that bilobectomy is a feasible operation with an oncological value. METHODS: From 1981 to 1998, 46 patients underwent bilobectomy for lung cancer. Eight upper and middle lobectomies (UML) and 38 middle and lower lobectomies (MLL) were performed. Intraoperative pneumoperitoneum was done in 11 MLL. We have considered operative mortality, postoperative complications, the persistence of drainage tubes and the length of hospital stay and the data were statistically compared with those relative to right lobectomies. Survival was estimated with the Kaplan-Meier method and the curves were compared with those of the right lobectomies and right pneumonectomies using the log-rank test. RESULTS: Overall morbidity was 43.4%. Mortality was 6.5%. Mean chest tube persistence was 7.8 days and mean hospital stay was 14 days. No statistical significance was found about these data comparing the UML and MLL separately and the bilobectomies with the right lobectomies. The pneumoperitoneum done in the MLL enabled a shorter hospital stay, statistically significant, in comparison with MLL without pneumoperitoneum. The overall 5-year survival rate was 38%. Considering the I and the II stages no statistical differences in survival were found considering the right lobectomies and right pneumonectomies. CONCLUSIONS: The bilobectomies can have a role in treatment of lung cancer that is equal to the other standard major resections. PMID- 11398045 TI - Pericardial mesothelioma following mantle field radiotherapy. AB - A 49-year-old female was referred with recurrent pericardial effusion following mantle field radiotherapy for Hodgkin s lymphoma. She underwent video-assisted thoracoscopy and resection of a pericardial window. Intraoperatively she suffered a cardiac arrest and subsequently died in the early postoperative period despite maximal therapy. Pathological examination revealed extensive myocardial fibrosis and multiple nodules of pericardial mesothelioma. The latter has hitherto not been recognised following mediastinal radiation. The cardiac complications of mantle field radiotherapy are discussed. PMID- 11398046 TI - Heparin management during cardiopulmonary bypass. PMID- 11398047 TI - Convective warming postbypass reduces postoperative bleeding. PMID- 11398048 TI - Aneurysm of the profunda femoris artery. PMID- 11398049 TI - [Diffusion-weighted echo-planar sequences for the evaluation of the upper abdomen: technique optimization]. AB - PURPOSE: To optimize the technique for the evaluation of molecular diffusion in the abdomen. MATERIAL AND METHODS: Fifteen healthy volunteers, 6 males and 9 females, ranging in age between 24 and 31 years underwent an MRI evaluation of the upper abdomen, using a superconductive 1.5T magnet (maximum gradient strength, 25 mT/m; minimum rise time 600 ms), equipped with phased array abdominal multicoil. Diffusion study was performed with a single-shot Inversion Recovery Spin-Echo Echo-planar sequence (IR-SE-EPI) with the following parameters: TR = infinite; TE=101 ms; matrix 128 yen 128; receiver bandwidth 2080 Hz/pixel; slices: n.20; slice thickness: 8 mm; acquisition time: 5.41 s. For diffusion weighting the following b values were employed: b=30 mm/s2, b=300 mm/s2 e b=500 mm/s2. Both qualitative and quantitative (calculation of linear regression analysis and of apparent diffusion coefficient) image analysis was performed. RESULTS: Image quality was graded as diagnostic in all the cases. Image quality decreased with the increase of b values: at low b values, the anatomy of upper abdominal organs was easily recognized, whereas, at high b values, the same organs could not be adequately assessed unless the images were compared with those obtained with low b values. Magnetic susceptibility artifacts were observed in all the cases; no significant chemical-shift artifacts were observed as the fat saturation pre-pulse was employed. Quantitative analysis demonstrated an apparent diffusion coefficient of 1.58 s/mm2 for the liver, 1.61 s/mm2 for the spleen and 5,14 s/mm2 for the gallbladder. A statistically significant difference (p<0.001) was observed between parenchymatous organs (liver and spleen) and gallbladder, presenting as a stationary fluid. CONCLUSIONS: Diffusion-weighted MR sequences may be implemented for abdominal studies, but the optimization of same parameters is slightly different compared with neuroradiologic applications. The potential applications are interesting above all as regards the characterization of focal liver lesions. Further developments are awaited in both sequence optimization (greater stability and lower sensitivity to magnetic susceptibility artifacts) and data analysis, with more complex algorithms able to better quantify the real diffusion coefficient. PMID- 11398050 TI - [Benign hepatic tumors: MRI features before and after administration of superparamagnetic contrast media]. AB - PURPOSE: To assess the yield of superparamagnetic iron oxide (SPIO)-enhanced MR images in the detection and characterization of benign hepatic tumors and to evaluate the potential role and safety of SPIO administration in the diagnosis of these tumors. MATERIAL AND METHODS: Eighteen patients underwent MRI before and after administration of SPIO particles. Spin echo (SE) T1, DP, T2 and Gradient echo (GE) T2* images were acquired with a.5 T superconductive unit. MR diagnosis was bioptically proved in 12 patients. In the remaining six patients, who had hemangiomas only, diagnosis was confirmed by at least two imaging techniques-such as MR, CT, ultrasonography, radio-labeled red cells scintigraphy-and by both clinical and imaging follow-up. RESULTS: Thirthy-four tumors were detected on the MR images: 29/34 (85,3%) before and 33/34 (97%) after SPIO administration - 6 focal nodular hyperplasias (FNH), 6 adenomas and 22 hemangiomas. One small tumor (adenoma) was detected on the unenhanced MR images only, while 4 lesions (3 adenomas, 1 FNH) were detected after SPIO administration only. DISCUSSION: SPIO enhanced MRI increased the detection rate of benign hepatic tumors compared to non-enhanced MRI. Iron oxide was also useful in the characterization of such lesions as it was able to demonstrate any heterogeneity resulting from the presence of central scars or septa. Nevertheless, in our experience it was useful to compare baseline with SPIO-enhanced MRI, even if time consuming. Indeed the uptake of iron oxide particles by well-differentiated lesions and normal hepatic parenchyma, is comparable, so that well-differentiated lesions appear isointense and therefore undetectable. CONCLUSIONS: In our experience, although numerically limited, SPIO-enhanced MRI was clinically safe and more effective than non enhanced MRI in both the detection and characterization of benign hepatic tumors, providing useful clues for diagnosis. PMID- 11398051 TI - [US and CT findings in complicated Meckel diverticulum]. AB - AIM: Meckel s diverticulum is the most common congenital abnormality of the small bowel. It may be silent or symptomatic when complicated by hemorrage, intestinal occlusion, diverticulitis and umbilical fistulas. Radiologic diagnosis is often difficult because abdominal plain radiography and ultrasound are not sufficiently specific; CT is most accurate in differential diagnosis. MATERIAL AND METHODS. 11 patients (age 5-69 ys) were studied. Clinical symptoms included acute abdomen (4 pts), intestinal occlusion (3 pts), abdominal pain (4 pts), fever (5 pts). Radiological studies were abdominal plain radiography (8 pts), ultrasound (5 pts), CT (9 pts). RESULTS: Abdominal plain radiography depicted signs of intestinal occlusion (4 pts) and perforation (1 pts); in 4 pts the signs were non diagnostic. Ultrasound showed an abscess in the pelvis (2 pts), dilatation and wall-thickening of an intestinal loop (2 pts), intestinal invagination (1 pts); it was not diagnostic in 3 pts. CT was not diagnostic in 3 pts; in 2 pts it showed an abscessual fluid collection in the pelvis, adherent to intestinal loops, with flogosis of the perivisceral fat; in 1 pt it revealed perforation; in 4 cases it was specific showing inversion of the diverticulum in an intestinal loop (2 pts) or a tubular fluid-filled structure, with thickened walls and contrast enhancement, which was interpreted as a inflammatory diverticulum (2 pts). DISCUSSION: Our series confirms the difficulty of diagnosing Meckel s diverticulum in an acute setting. Abdominal plain radiography only allowed to diagnose intestinal occlusion or perforation. Ultrasound revealed abscessual collections in the pelvis, fluid distention of the diverticulum, segmental thickening of the intestinal walls and invagination. CT proved to be more specific showing signs suggestive of correct diagnosis in 6 pts. In particular, evidence of an intraluminal prolonged mass with central area of fat density and peripherral collar was considered suggestive of intraluminal invagination of Meckel s diverticulum. Another diagnostic sign is the evidence of a tubular fluid filled structure, with thickened, enhanced walls. In 2 cases CT showed an abscessual collection with gas-fluid level (complication of perforation) confirming the need for surgery. CONCLUSION: Meckel s diverticulum is a not uncommon condition that in some cases is complicated, resulting in acute abdomen. Preoperative radiological diagnosis can be suspected in the presence of suggestive signs, more often depicted by CT. PMID- 11398052 TI - [Usefulness of CT colonography in the preoperative evaluation of patients with distal occlusive colorectal carcinoma]. AB - PURPOSE: To evaluate the role of preoperative virtual colonoscopy to study the proximal colon in patients with distal occlusive carcinomas, diagnosed by conventional colonoscopy. MATERIAL AND METHODS: We examined 19 patients aged 46 to 83 years (13 men and 6 women) with distal occlusive colorectal carcinomas diagnosed by conventional colonoscopy, who were preoperatively studied with virtual colonoscopy. Patients with acute bowel obstruction were excluded. Results were compared with the findings of preoperative conventional colonoscopy and barium enema examination, intraoperative colon palpation, histopathologic outcome, postoperative conventional colonoscopy and barium enema examination. RESULTS: Virtual colonoscopy identified all 19 distal occlusive colon carcinomas and 22 synchronous lesions, 2 cancers (prevalence 10,6%) and 20 polyps (prevalence 68,4%). Both synchronous cancers were confirmed intraoperatively by direct palpation. Postoperative conventional colonoscopy, which was performed in 18 patients, confirmed the presence of 15 polyps in 12 patients. Three subcentimeter polyps were removed during conventional colonoscopy and were missed at virtual colonoscopy. Two polyps shown by virtual colonoscopy were not found at conventional colonoscopy. Postoperative barium enema was performed in three patients and confirmed three polyps identified at virtual colonoscopy. Preoperative barium enema was performed in five patients and failed to adequately demonstrate the proximal colon. Virtual colonoscopy showed a sensitivity of 87% and a specificity of 75%. CONCLUSIONS: Virtual CT colonoscopy can be considered an important diagnostic technique to evaluate preoperatively the proximal colon in patients with distal occlusive carcinomas,as it gives better results than barium enema or conventional colonoscopy, as well as being well tolerated and less invasive. PMID- 11398053 TI - [Role of color Doppler US in the evaluation of renal transplant]. AB - AIM: To assess the role of Ultrasound (US), US Color Doppler (CD) and Power Doppler (PD) in the diagnosis and in the follow-up of renal graft pathology by evaluating morphological and functional features of the vasculature and comparing these to other clinical parameters. MATERIAL AND METHODS: From January 1990 to June 2000, four hundred and thirty-six renal allograft recipients (mean age 45 years) underwent periodical US, CD and PD (mean follow-up 48 months) to evaluate morphology and perfusion of the graft. Resistive index (RI) and pulsatility index (PI) were measured in order to monitor flow variations from the renal to the arcuate arteries. PD was used mainly to study the morphology of the cortical vessels. The examinations were performed on an Esaote Biomedica AU-4/5 (Genoa, Italy) using a convex 3.5 MHz probe and a linear 7.5 MHz probe. On the basis of clinical data the patients were divided into 3 groups: A) Normal, B) Acute graft dysfunction, C) Chronic allograft nephropathy. In 87 patients (20%) percutaneous biopsy or FNAB was performed. RI and PI mean values +/-SD were calculated and compared to the other diagnostic parameters considered: serum creatinine level, US morphology, CD and PD vascularization. Finally RI and PI for each group were compared using the t -test in order to determine the statistical significance of the correlation between these indices and the patients clinical conditions. RESULTS: The 436 patients were divided as follows: Group A) 170 patients (39%); Group B) 105 patients (24%); Group C) 161 patients (37%). Urological and surgical complications were ruled out in all patients. RI and PI showed a similar trend exceeding cut-off values in Group B) and C) with highest peaks in Group B. Statistical analysis demonstrated the efficacy of this method in the differentiation between normal and pathological grafts, but there was a reduced statistical difference between the two pathological groups. Histological analysis performed on 87 patients (20%) showed good correlation with RI. DISCUSSION AND CONCLUSIONS: CD is a non-invasive diagnostic method which provides flow-metric quantitative parameters for the hemodynamic assessment of the renal transplant. These values present a certain sensitivity but are not specific of renal graft dysfunction, as there is no reliable differentiation between acute rejection and other parenchymal pathologies. During the follow-up, RI and PI have no predictive value. RI variations from renal artery to cortical vessels (hylum-cortical ratio) show a good correlation with the clinical evolution of the transplant. The evaluation of RI and PI can generally be limited to renal and interlobar arteries as arcuate sampling is necessary only when the hylum-cortical ratio shows reduction or inversion. Integration of clinical and instrumental diagnoses can reduce the number of biopsies. PMID- 11398054 TI - [Acute pyelonephritis. Role of diagnostic imaging]. AB - PURPOSE: To asses the role of imaging in acute pyelonephritis by retrospectively evaluating a number of cases observed with special care to US and CT signs that may be useful for an early and correct diagnostic assessment. MATERIAL AND METHODS: From 1994 to the first quarter of 2000 we observed 95 patients clinically suspected of having acute pyelonephritis (81 females and 14 males, age range 2-80 years; 75% were under 40 years old). Sonography was performed as an emergency in 78 patients. CT scan with contrast media was performed in acute phase in 78 patients acquiring the images in the pre-contrastographic phase, 60 s after the administration of contrast media (portal phase), after 5 to 10 min (excretory phase), and after 3 to 6 hours (very late phase). In 47 patients, diagnosed as having acute pyelonephritis, a follow-up CT scan was performed at 30 days, 45 days, 3 months and 6 months from the acute episode. RESULTS: In 78 of the patients clinically suspected of having acute pyelonephritis, the CT examination allowed to assess the presence or lack of the inflammatory focus. In 17 cases US discovered causes other than acute pyelonephritis on admission; in 25 out of 61 patients it allowed assessment of a pyelonephritic focus that was confirmed with CT. The CT examination with contrast media proved to be a reliable and fondaments method in the diagnosis of pyelonephritic focus especially as it revealed the lack of concentration of contrast media in the flogistic foci, in the very late fase. This method furthermore allows to discover alterations in peri and pararenal regions and to rule out abscesses and renal infarcts. US had a sensitivity of 50% and a specificity of 70% in the diagnosis of acute pyelonephritis, but those may be improved by power Doppler. DISCUSSION AND CONCLUSIONS: In suspected acute pyelonephritis US offers useful indications for a quick and correct diagnostic assessment, but CT examination with contrast media proved to be more reliable than US. In our experience US and above all CT examination rapidly identified patients affected by acute pyelonephritis, thereby allowing us institute a timely antibiotic therapy and obtain a brilliant therapeutic response. PMID- 11398055 TI - [Popliteal cysts: sonography evaluation after arthroscopic treatment]. AB - PURPOSE: To identify a set of sonomorphological parameters to evaluate the outcome of arthroscopic surgery for popliteal cysts by sonography. MATERIAL AND METHODS: We examined 30 patients (18 males, 12 females) before treatment and 3/6 months after arthroscopy. Patients were classified by clinical (4 groups) and sonographic criteria. The sonographic criteria were the morphological classification of the cysts (beak-shaped, X-shaped, grape-like, slit-shaped) and the volume measurement. RESULTS: Sonography identified beak-shaped popliteal cysts (the most frequent Baker cyst shape), 5 grape-like (the usual shape of complicated Baker cysts) and 1 X-shaped . The volume of cysts before treatment ranged from 5 to 38 cl. Most cysts were clinically rated as grade 2 (symptomatic and evident at physical examination, with a reduction of range of motion up to 20 degrees of flexion). No patient was rated as grade 0 (normal). After surgery the cyst was still evident at sonography. After 3 months we recorred a cyst volume decrease in all cases. Six months after surgery 29/30 cysts had a volume <5 cl with an almost virtual cavity that changed their appearance in to slit-shaped . In one case cyst volume increased. The clinical classification showed an increase of the grade 0 group unsymptomatic cysts (not detectable at physical examination). DISCUSSION AND CONCLUSIONS: Sonographic evaluation of popliteal cysts after arthroscopic surgical treatment allowed to recognize the cysts, which are not removed by this approach. The follow-up at 3 months showed a reduction in cyst volume. After six months, in the cases with a good surgical response, the cysts reached a volume <5 cl regardless of their original volume. The morphology of the cysts with a good surgical result, is slit-shaped with virtual cavity regardless of their original appearance. There is a high correlation between sonographic and clinical data. We believe that the volume reduction of the cyst three months after surgery is a good predictive element. After six months the preesence of a cyst with a volume <5 cl and a slit-shape with virtual cavity is a sign of good surgical result. PMID- 11398056 TI - [Normal anatomy and pathological conditions of subscapular muscle: US findings compared with surgery]. AB - PURPOSE: To compare US results with those of surgery in the assessment of the subscapularis tendon. MATERIAL AND METHODS: From January 1995 to December 1998 1500 patients underwent US of the shoulder. 12 of these patients had an injured subscapularis tendon. US results of these patients were evaluated retrospectively and compared with those of surgery. US examinations of the subscapularis tendon were performed with two US units (AU4 Idea and HDI 3000), using linear arrays 10 13 MHz transducers. The following features of the tendon were evaluated: morphology, thickness and echotexture. RESULTS: A) US findings. US showed: - chronic degenerative tendon changes with diffusely inhomogeneous echotexture (5 cases); - focal tears with small hypoechoic ill-defined areas or gross hypo anechoic areas involving the full thickness of the tendon (4 cases); - complete tendon tear with disappearance on US of the subscapularis tendon or with severe thickening of the tendon (2 cases); - in 1 case of recent anterior dislocation of the shoulder a bulky hematoma was seen with coexisting thickening and subtotal tear of the tendon. B) Surgical findings. US findings were confirmerd at surgery in 10 of the 12 patients. In the patient with anterior dislocation of shoulder the diagnosis hematoma was confirmed, but the subscapularis tendon was undamaged. Moreover, in 1 out of the 5 patients with US diagnosis of chronic degenerative tendon changes, a small focal tear of the tendon was found at surgery. CONCLUSION: US is a first level investigation which is able to demonstrate early changes of the subscapularis tendon and nearby structures. Awareness of pitfalls might decrease the use of second level investigations such as CT arthrography and MR imaging, which on the other hand allow a better definition and assessment of the extent of the injury. PMID- 11398057 TI - [Virtual laryngoscopy of neoplastic pharyngeal and laryngeal pathology]. AB - PURPOSE: This study investigated the feasibility and clinical value of high resolution virtual laryngoscopy (VL) based on helical CT data sets. MATERIAL AND METHODS: Fifteen patients with neoplastic laryngeal pathology underwent helical CT examination performed with controlled breath. The CT data were downloaded to a workstation equipped with software for three-dimensional rendering (Silicon Graphics O2, Vitrea Visualization 1.2.8). Two radiologists using Fly Through software generated both antegrade and retrograde endoluminal virtual endoscopy navigations of the larynx. The results of the radiological study were compared with conventional endoscopy findings. RESULTS AND CONCLUSIONS: Virtual laryngoscopy provided the correct diagnosis in 13 of 15 cases. VL is capable of simulating the visual findings of endoscopy in cases of laryngeal tumors and subglottic stenosis. Small and plane tumors of the vocal chords are not adequately visualized. This technique appears to be useful both as biopsy guide during conventional laryngoscopy and as a correct pre-surgery staging, particularly in stenotic tumors. PMID- 11398058 TI - [Ocular ultrasonography in pediatrics: persistence of hyperplastic primary vitreous]. AB - AIM OF THE STUDY: To assess the diagnostic viability of ultrasound and color Doppler ultrasound in a particular segment of paediatric ophthalmology-persistent primary hyperplastic vitreous, in the presence of leucocoria, retrospectively evaluated in patients observed over the last two years. MATERIAL AND METHODS: We re-evaluated four patients (two new-born, one unweaned and one nine-years-old) who at ophthalmoscopic examination were suspected having persistent primitive hyperplastic vitreous. The follow-up included an ultrasound examination, basic color-Doppler ultrasound and re-evaluation during sedation both using an operative microscope and ultrasound. The examinations were performed with linear 7.5 Mhz probes and a Doppler frequency of 3.7 Mhz, while the sedation examination was performed with an anular 13 Mhz probe. The scans were sagittal and axial to correctly localize the lesion with respect to the lens, to ciliary bodies and to the optic nerve head. RESULTS: In the first patient ultrasound revealed a hyperechogenic inhomogeneous structure bilaterally in the vitreous structure; this extended from the posterior wall of the lens to the optic nerve head and retina, and was found to be highly vascolarized at the subsequent color-Doppler ultrasound. In the second patient there was an echogenic band extending from the posterior wall of the lens to the optic nerve head and to the retina, as the fourth patient showed a series of echogenic bands extending from the temporal ciliar of bodies to the temporal retina; in both cases no significant vascolarization was found by color Doppler ultrasound. In the third patient ultrasound showed a lesion involving nearly all of the vitreous body, with inhomogeneous structure with small calcifications and intense vascolarization. DISCUSSION AND CONCLUSIONS: Ultrasound supplies essential information for the diagnosis of persistent hyperplastic primary vitreous as it determines the presence of the lesion, its extension and retinal and optic nerve head involvement. As confirmed by operative microscopy the first patient had bilateral lesions involving the optic nerve head, as well as the retina, the latter which appeared raised. The second and fourth patients had typically posterior lesions; the lesion of the third patient was difficult to interpret even by operative microscopy. To conclude,Ultrasound proved to have an optimal sensitivity and specificity for precisely locating the site and extension of the lesion. By contrast, the difficulties and need for sedation with color Doppler ultrasound unlikely to be useful with paediatric patients. PMID- 11398059 TI - [Computed tomography evaluation of metastasis of cutaneous melanoma]. AB - PURPOSE: To assess the value of Computed Tomography (CT) in the diagnosis and in morphologic characterization of metastatic melanoma. MATERIALS AND METHODS: The data of total body CT of 124 consecutive patients with melanoma having a Breslow index 1 mm or a positive sentinel lymph node have been retrospectively reviewed. RESULTS: The CT scan showed loco-regional and/or distant metastases in 36 patients (39%). Ten of these (28%) had metastases only to lymph nodes, whereas 26 patients (72%) had multiple metastases. Nodal, pulmonary, brain, subcutaneous, hepatic, adrenal, bone, gastrointestinal, breast and abdominal wall metastases were detected in 80.6%, 47.2%, 25%, 25%, 16.7%, 13.9%, 11.1%, 5.6%, 5.6% and 2.8% of the patients respectively. All the patients with metastases also had a positive sentinel lymph nodes and/or symptoms of metastatic disease. CONCLUSIONS: CT fails to reveal any characteristic feature of metastatic melanoma, but it is of value in the diagnosis of loco-regional and distant metastases in III stage disease. PMID- 11398060 TI - [Diagnostic imaging and patient database managing systems: The integration of digital information in the experience of an intensive care center]. AB - AIM: To present our experience with integrating digital information on Intensive Care Unit patients (clinical data, laboratory findings, imaging, etc) to create electronic patient records. MATERIAL AND METHODS: Using the hospital Intranet, a connection was established between the Local area Network (LAN) of the Intensive Care Unit (ICU) and the Digital Imaging and Communications in Medicine (DICOM(R)) network of the Radiology Department allowing to receive, process and archive digital images locally at the ICU. Using the software RADclient-RADimage, the information received was managed by an electronic patient record system (DIGISTAT by UMS-Unterberger Medical Software, Florence). All the above software runs on Microsoft WindowsNT 4.0 platforms. RESULTS: Images of various kinds and formats (CT, MRI, etc.) pertaining to the ICU patients were semi-automatically handled and filed on a local server acting as a central databank. The images were then included in the electronic patient record and made available to the end user who could view them using either web technologies (hypertexts were automatically generated that could be viewed through the widely available World Wide Web browsers) or specific viewing utilities supplied with DIGISTAT . DISCUSSION AND CONCLUSIONS: For the intensivist, the handling and filing of data on hospitalised or discharged patients for treatment or research purposes involves having to process large amounts of information. Furthermore, in the event of patients being re-admitted to the unit, it is crucial to have ready access to all the information regarding previous hospital stays, including diagnostic images, to avoid the need for time-consuming searches through the hospital s paper-based archives. The possibility to access clinical information and diagnostic images using a single computer programme proved to be useful both for evaluating the patient s conditions immediately after the imaging procedure and for monitoring the patient s progress over time by comparing the different diagnostic images and imaging procedures. This pilot experience could be seen to provide the basic know how for applying the method in the future Emergency Department of the A. Gemelli Hospital in Rome. PMID- 11398061 TI - [Strange but true: acetylcysteine protects against kidney damage caused by iodized contrast media]. PMID- 11398062 TI - [Magnetic Resonance Imaging in Von Recklinghausen s neurofibromatosis. Value of target sign in 2 cases]. PMID- 11398063 TI - [Intrathoracic meningocele in von Recklinghausen s neurofibromatosis. A case report]. PMID- 11398064 TI - [Biliary obstruction and arterialization phenomena. Observations of 4 cases studied with spiral computed tomography]. PMID- 11398065 TI - [Rudimentary acral anomalies in a patient with Acrocal-losal syndrome]. PMID- 11398066 TI - [A rare case of an isolated fracture of the acetabulum in a patient with congenital coxa vara]. PMID- 11398067 TI - [Radiologic and MR findings in a rare case of intermediate cuneiform osteonecrosis in a child]. PMID- 11398069 TI - Inhaled corticosteroids: impact on asthma morbidity and mortality. AB - Inhaled corticosteroids are now recommended as first-line therapy for asthma. Although these drugs clearly improve the symptoms of the disease and the associated physiologic abnormalities, epidemiologic studies provide important information on their effectiveness in preventing asthma morbidity and mortality. We review the evidence regarding the role of inhaled corticosteroids in the prevention of asthma fatality and hospitalization. In the process, we discuss the methodologic complexities of the nonexperimental studies and the implications of the methodologic issues on the evaluation of the impact of these drugs. Eight of the cohort and ecologic studies conducted to date strongly suggest that inhaled corticosteroids, when taken regularly, decrease the number of hospitalizations for asthma by up to 80%. For asthma death, the results of 11 investigations appear less consistent, especially those of several cohort and case-control studies whose principal objective was to examine not the benefit of inhaled corticosteroids but the adverse effects of other drug classes. Much of the inconsistency in the results, however, can be explained by weaknesses in study design and analysis-in particular, the failure to consider exposure in terms of regular use of inhaled corticosteroids. When the most recent study involving the use of the Saskatchewan databases is considered, it is evident that regular treatment with conventional or low-dose inhaled cortico-steroids results in a significant reduction in fatalities due to asthma. In all, the evidence to date strongly indicates that regular use of inhaled corticosteroids, even at low doses, would prevent the major portion of asthma hospitalizations and deaths. PMID- 11398070 TI - Pulmonary T cells and eosinophils: coconspirators or independent triggers of allergic respiratory pathology? AB - Etiologic discussions of allergic respiratory pathology frequently engender rabid constituencies of pro-T cell or proeosinophil disciples, each claiming, often with religious fervor, the importance of their leukocyte. However, increasing evidence suggests that the exclusionary rhetoric from either camp is inadequate to explain many of the pathologic changes occurring in the lung. Data from both asthmatic patient and mouse models of allergic respiratory inflammation suggest that, in addition to cell-autonomous activities, T-cell and eosinophil interactions may be critical to the onset and progression of pulmonary pathology. These studies also suggest that T-lymphocyte subpopulations and eosinophils communicate by means of both direct cell-cell interactions and through the secretion of inflammatory signals. Collectively, the data support an expanded view of T-cell and eosinophil activities in the lung, including both immunoregulative activities and downstream effector functions impinging directly on lung function. PMID- 11398071 TI - Natural rubber pharmaceutical vial closures release latex allergens that produce skin reactions. AB - BACKGROUND: The release of allergenic proteins from natural rubber vial closures (stoppers) into aqueous pharmaceuticals may induce allergic reactions in individuals with latex allergy (LA) receiving medications from such vials. OBJECTIVE: The goal of this study was to determine whether solutions stored in vials containing natural rubber closures release allergenic proteins detectable by skin testing of subjects with LA. METHODS: Five pharmaceutical vial closures (2 natural rubber and 3 synthetic) were coded, inserted onto vials containing phenol-saline-human serum albumin, and stored in an inverted position before use. Twelve volunteers with and 11 volunteers without LA underwent skin testing with solutions from each of the 5 vials, either those not punctured (0P) or those punctured 40 times with a 21-gauge needle 12 to 24 hours before testing (40P). RESULTS: All intradermal skin test responses in the group without LA were negative. Two and 5 of the 12 subjects with LA had positive intradermal skin reactions to 0P and 40P solutions, respectively, from vials containing rubber closures. Two subjects with LA had inexplicable, positive, nonreproducible intradermal skin test reactions to solutions from vials containing bromobutyl but not vials with isoprene synthetic closures. In vitro inhibition analysis detected 6 to 7 AU/g latex allergen in extracts of cut natural rubber containing closures but not in extracts of synthetic closures. CONCLUSION: Natural rubber vial closures released allergenic latex proteins into the tested solutions in direct contact during storage in sufficient quantities to elicit positive intradermal skin reactions in some individuals with LA. These data support a recommendation to eliminate natural rubber from closures of pharmaceutical vials. PMID- 11398072 TI - Efficacy of soluble IL-4 receptor for the treatment of adults with asthma. AB - BACKGROUND: IL-4 mediates important proinflammatory functions in asthma, including induction of the IgE isotype switch, increased expression of vascular cell adhesion molecule 1 and promotion of eosinophil transmigration across the endothelium, stimulation of mucus production, and T(H)2 lymphocyte differentiation, leading to release of IL-4, IL-5, IL-9, and IL-13. OBJECTIVE: The current study evaluated the therapeutic potential of inhaled recombinant human soluble interleukin-4 receptor (IL-4R) as an IL-4 antagonist. METHODS: This study was a randomized, double-blind, placebo-controlled study in 62 subjects involving 12 once weekly nebulizations of 0.75, 1.5, or 3.0 mg of IL-4R or placebo. During screening, subjects documented dependence on inhaled corticosteroids by an exacerbation in asthma induced by one or two 50% dose reductions at 2-week intervals. After restabilization for 2 weeks on the dose above which their asthma flared, inhaled steroids were discontinued, patients were randomized, and study medication was started on day 0. RESULTS: IL-4R was well tolerated. Efficacy was demonstrated by a decline in FEV(1) observed in the placebo group (-0.4 L and -13% predicted), which did not occur in the group receiving 3.0 mg of IL-4R (-0.1 L and -2% predicted; P =.05 over the 3-month treatment period). Daily patient-measured morning FEV(1) also demonstrated a significant decline in the placebo group (-0.5 L and -18% predicted), which did not occur in the group receiving 3.0 mg of IL-4R (-0.1 L and -4% predicted; P =.02 over the 3-month treatment period). The efficacy of IL-4R was further confirmed by the absence of increase in asthma symptom scores in the group receiving 3.0 mg of IL-4R (Delta 0.1) compared with that seen in the placebo group (Delta 1.4 over 1 month; P =.07). Study discontinuation for asthma exacerbation was not significantly different between groups (placebo, 56%; 3.0 mg of IL-4R, 47%; P = not significant). CONCLUSION: These promising data suggest that IL-4R is safe and effective in the treatment of moderate persistent asthma. PMID- 11398073 TI - Grass pollen immunotherapy: symptomatic improvement correlates with reductions in eosinophils and IL-5 mRNA expression in the nasal mucosa during the pollen season. AB - BACKGROUND: Tissue eosinophilia and infiltration by T(H)2-type T cells are characteristic features of allergic rhinitis both after allergen challenge and during natural allergen exposure. Specific immunotherapy inhibits allergen induced nasal eosinophilia. OBJECTIVES: We sought to assess, in the context of a randomized trial, the relationships between symptomatic improvement after immunotherapy and eosinophil numbers and IL-5 expression in the nasal mucosa during the pollen season. METHODS: Nasal biopsy specimens were taken from 37 adults with severe summer hay fever at baseline (out of season) and at peak season after 2 years of treatment with a depot grass pollen extract or placebo. Biopsy specimens were processed for immunohistochemistry by using mAbs against eosinophils (EG2), T cells (CD3), and IL-2 receptor-positive cells (CD25), as well as for in situ hybridization by using a sulfur 35-labeled antisense riboprobe directed against IL-5. RESULTS: Immunotherapy significantly reduced symptoms (49%, P =.01) and medication requirements (80%, P =.007) compared with placebo. There was a 400% increase (P =.004) in eosinophils during the pollen season in placebo-treated patients, which was inhibited in the immunotherapy group (20% increase, P =.04 between groups). Seasonal increases were also observed for CD25(+) cells (P =.002), CD3(+) cells (P =.02), and IL-5 mRNA expressing cells (P =.03) in the placebo group but not in the immunotherapy group. A significant correlation was observed between eosinophils and IL-5 expression (r = 0.5, P <.05). Both eosinophils (r = 0.6, P <.02) and IL-5 (r = 0.6, P <.02) correlated with symptoms after immunotherapy. CONCLUSION: Improvement in symptoms after grass pollen immunotherapy may result, at least in part, from inhibition of IL-5-dependent tissue eosinophilia during the pollen season. PMID- 11398074 TI - The molecular basis of antigenic cross-reactivity between the group 2 mite allergens. AB - BACKGROUND: Mite group 2 allergens Der p 2, Der f 2, and Eur m 2 are 14-kDa proteins of unknown function that share 83% to 85% amino acid sequence identity. Isoforms of the allergens within each genus have been identified which differ by 3 or 4 amino acids, but little is known of the influence of group 2 polymorphisms on human IgE antibody binding. OBJECTIVE: The purpose of this study was to investigate the importance of interspecies and isoform substitutions on murine mAb and IgE antibody binding and on the molecular structure of the group 2 allergens. METHODS: Site-directed mutagenesis was used to incorporate the isoform amino acid substitutions onto the Der p 2.0101 sequence. Recombinant allergens were expressed and purified from Escherichia coli and used to evaluate antibody binding by enzyme-linked immunosorbent assay (ELISA). Molecular modeling of the tertiary structure was used to analyze structural differences between the various group 2 allergens. RESULTS: The substitution of asparagine for aspartic acid at position 114 restored mAb binding of rDer p 2.0101; the other Der p 2 isoforms and the 3 rDer f 2 isoforms also reacted in the 2-site ELISA. The correlation of IgE binding to the Der p 2 isoforms was excellent and tended to be higher in the isoforms with the asparagine 114 substitution (r (2) = 0.87 vs r (2) = 0.95). rEur m 2.0101 bound to all mAb except 7A1; when compared with rDer p 2 for IgE binding, rEur m 2.0101 gave a correlation coefficient of r (2) = 0.68. Molecular modeling revealed that Eur m 2 and the storage mite homologs Lep d 2 and Tyr p 2 retain the tertiary fold of Der p 2. Eur m 2 has a conserved surface, whereas Lep d 2 and Tyr p 2 present most of the amino acid substitutions on this surface. Lep d 2 and Tyr p 2 did not react with mAb or with sera from patients with IgE to Dermatophagoides species. CONCLUSION: The isoform substitutions of rDer p 2 can be distinguished by mAb. The allergenic cross-reactivity between Der p 2, Der f 2, and Eur m 2 is a direct result of the conserved antigenic surface, whereas the lack of cross-reactivity with Lep d 2 and Tyr p 2 is a result of the multiple substitutions across this surface. PMID- 11398075 TI - Allergens of wild house dust mites: environmental Der p 1 and Der p 2 sequence polymorphisms. AB - BACKGROUND: Sequence diversity is a common feature of mite allergens. Previous studies, using predominantly commercial mite clones, have described several polymorphic residues for Der p 1 and Der p 2. OBJECTIVE: This study aimed at determining the occurrence of sequence diversity in environmental mite isolates. METHODS: Mites were isolated from houses in Perth and Sydney, Australia. Total RNA was extracted from 1 to 30 Perth mites, and cDNA was synthesized by reverse transcriptase PCR. Der p 1 and Der p 2 cDNAs were PCR amplified and sequenced. Genomic Der p 1 DNA was amplified from whole Sydney mites directly by PCR and then sequenced. RESULTS: Twelve Der p 1 and 9 Der p 2 cDNA clones and 3 Der p 1 genomic DNA were analyzed and showed a high frequency of amino acid polymorphisms. Der p 2 displayed a clear pattern of divergence toward 2 alleles that differed by 4 amino acids and had characteristic silent nucleotide changes. The pattern for Der p 1 was different and unusual, with almost no silent nucleotide substitutions but frequent sporadic missense changes. Proliferative responses of peripheral blood mononuclear cells to peptides containing polymorphic residues of Der p 1 were detected in 8 of 19 subjects, with stimulation being found only for either one of the variant forms of the peptides. However, the responses to variants of whole recombinant allergens were similar, as shown for 4 variants of Der p 2. CONCLUSION: Two clones for each of the allergens were identified as containing sequences that were largely representative of environmental isolates. A small-scale reverse transcriptase PCR used to produce cDNA from individual mites isolated from house dust will have wide application for studies on mite genetics and the production of recombinant mite allergens. Differences in T-cell responses to peptides representing variant epitopes were found, but responses to variants of whole recombinant allergens were similar. The GenBank and Swiss Prot database entries for Der p 1 (U11695) and Der p 2 (P49278) have been updated with the inclusion of the sequence polymorphisms described in this study. PMID- 11398076 TI - Human eosinophils induce histamine release from antigen-activated rat peritoneal mast cells: a possible role for mast cells in late-phase allergic reactions. AB - BACKGROUND: Mast cells and eosinophils are believed to interact during the late and the chronic stages of allergic inflammation. OBJECTIVE: In this study we investigated whether eosinophils can cause activation and consequent histamine release of already challenged mast cells, a situation likely to take place during the allergic late-phase reaction. METHODS: Rat peritoneal mast cells presensitized with IgE anti-dinitrophenol-human serum albumin and challenged by dinitrophenol-human serum albumin or compound 48/80 were incubated with either eosinophil sonicate or major basic protein (MBP). Eosinophils were purified from the peripheral (>98%) blood of mildly allergic patients. Heparin and pertussis toxin and different extracellular Ca(2+) concentrations were used to modulate mast cell reactivation by MBP. Histamine release was assessed as a marker of mast cell activation. RESULTS: IgE-challenged mast cells were sensitive to reactivation induced by eosinophil sonicate and MBP. Reactivation was not cytotoxic for the mast cells. Mast cells previously challenged with compound 48/80 did not respond to subsequent MBP activation. Furthermore, heparin and pertussis toxin both inhibited mast cell reactivation induced by MBP. The ability of eosinophil sonicate and MBP to activate mast cells was not significantly affected at the different Ca(2+) concentrations. CONCLUSIONS: In summary, we have shown a direct activating activity of eosinophils, partially due to MBP, toward IgE-challenged and immunologically desensitized mast cells. This suggests that in vivo mast cells can be reactivated during a late-phase reaction to release histamine by a non-IgE-dependent mechanism. PMID- 11398077 TI - IL-4 and IL-13 induce myofibroblastic phenotype of human lung fibroblasts through c-Jun NH2-terminal kinase-dependent pathway. AB - BACKGROUND: Myofibroblasts play a role in the airway remodeling response of bronchial asthma. IL-4 and IL-13 are possibly involved in the airway remodeling response by inducing extracellular matrix production by fibroblasts. However, the roles of these cytokines in inducing the phenotypic modulation of human lung fibroblasts (HLFs) to myofibroblasts and the intracellular signal have not been determined. OBJECTIVE: We examined the effect of IL-4 and IL-13 on inducing the phenotypic modulation of HLFs to myofibroblasts characterized by alpha-smooth muscle actin and examined the role of the mitogen-activated protein (MAP) kinase superfamily in inducing the myofibroblastic phenotype of the HLF to clarify these issues. METHODS: Phosphorylation and activities of c-Jun NH(2)-terminal kinase (JNK), p38 MAP kinase, and extracellular signal-regulated kinase (Erk) were examined by using Western blotting and in vitro kinase assay. Expression of alpha smooth muscle actin in IL-4- and IL-13-stimulated HLFs was analyzed by means of Western blotting. RESULTS: The results showed that (1) IL-4 and IL-13 increased alpha-smooth muscle actin expression in a dose- and time-dependent manner; (2) IL 4 and IL-13 induced increases in JNK and Erk phosphorylation and activity but not p38 MAP kinase activity; (3) CEP-1347 and PD 98059 attenuated IL-4- and IL13 induced JNK and Erk activity, respectively; and (4) CEP-1347, but not PD 98059, attenuated IL-4- and IL-13-induced alpha-smooth muscle actin expression. CONCLUSION: These results indicate that IL-4 and IL-13 are capable of inducing the phenotypic modulation of HLFs to myofibroblasts, and JNK, but not p38 MAP kinase and Erk, regulates IL-4- and IL-13-induced phenotypic modulation of HLFs to myofibroblasts. PMID- 11398078 TI - Expression of the high-affinity IgE receptor on peripheral blood dendritic cells: differential binding of IgE in atopic asthma. AB - BACKGROUND: Dendritic cells can express the high-affinity IgE receptor (FcepsilonRI), which, in the presence of specific IgE, facilitates the uptake of allergen, leading to increased activation of allergen-specific T cells. FcepsilonRI expression by dendritic cells is higher in the airways of atopic asthmatic subjects than in those of healthy, nonatopic control subjects. OBJECTIVE: The aims of this study were to determine whether a similar difference in FcepsilonRI expression occurs between dendritic cells in the peripheral blood of atopic asthmatic subjects and healthy individuals and also whether an altered ability of FcepsilonRI(+) peripheral blood dendritic cells to bind IgE accompanies the atopic asthmatic state. METHODS: Flow cytometry was used to analyze the surface expression of FcepsilonRI and exogenously bound IgE on dendritic cells identified as lineage negative (CD3, CD14, CD16, CD19, and CD56) and HLA-DR bright. RESULTS: The total expression of FcepsilonRI on the surface of dendritic cells from healthy and asthmatic subjects was not significantly different. However, in vivo, dendritic cells from atopic asthmatic subjects had higher levels of receptor occupancy by IgE and bound exogenous IgE in vitro more efficiently than dendritic cells from healthy subjects. CONCLUSION: The similar levels of expression of FcepsilonRI on peripheral blood dendritic cells from healthy and asthmatic subjects suggest that the local environment in the airway is responsible for the upregulation of surface FcepsilonRI on airway dendritic cells in asthma. The results also suggest that the functional ability of FcepsilonRI to bind IgE is differentially controlled in the atopic state. PMID- 11398079 TI - L-Selectin is required for the development of airway hyperresponsiveness but not airway inflammation in a murine model of asthma. AB - BACKGROUND: Airway inflammation and airway hyperresponsiveness (AHR) are fundamental features of asthma. Migration of inflammatory cells from the circulation into the lungs is dependent on adhesion molecule interactions. The cell surface adhesion molecule L-selectin has been demonstrated to mediate leukocyte rolling on inflamed and noninflamed pulmonary endothelium. However, its role in the development of airway inflammation and AHR in asthma has not been examined. OBJECTIVE: We sought to characterize the role of L-selectin in the recruitment of inflammatory cells to the airway-lung and the development of AHR in a murine model of asthma. METHODS: An ovalbumin (OVA)-induced allergic airway disease model of asthma was applied to L-selectin-deficient (LKO) mice and C57BL/6 wild-type (WT) control mice. The development of airway inflammation was assessed by examining leukocyte influx into bronchoalveolar lavage (BAL) fluid and the lung. Total and differential BAL leukocyte counts were determined, and the immunophenotype of BAL lymphocytes was assessed by means of flow cytometry. The development of AHR was assessed by means of whole-body plethysmography. RESULTS: Airway-lung inflammation was equivalent in LKO and WT mice sensitized challenged with OVA, as measured by total and differential BAL cell counts and histologic analysis of lung tissue. Numbers of eosinophils, neutrophils, lymphocytes, and monocytes in BAL fluid were equivalent in LKO and WT mice. However, phenotypic analysis of BAL lymphocytes demonstrated significantly reduced CD3(+) populations and increased B220(+) populations in LKO compared with WT mice (P <.05). Remarkably, despite a fulminant inflammatory response in the airway-lung in LKO mice sensitized-challenged with OVA, AHR was completely abrogated. CONCLUSION: L-selectin plays a crucial role in the development of AHR but not allergic inflammation in an animal model of asthma. L-selectin represents a potential target for novel asthma therapies specifically aimed at controlling AHR. PMID- 11398080 TI - Anti-IgE efficacy in murine asthma models is dependent on the method of allergen sensitization. AB - BACKGROUND: Murine models used to delineate mechanisms and key mediators of asthma have yielded conflicting results and suggest that the dominant mechanism and mediators required for disease induction differ depending on the model and method of allergen sensitization used. OBJECTIVE: The goal of this study was to determine whether the mode of allergen sensitization influenced the role that IgE had in allergen-induced pulmonary eosinophilic inflammation. METHODS: Mice were exposed to dust mite extract in 2 models of allergic inflammation that differed in the method of sensitization. We compared sensitization by aerosol exposure with and without concomitant human respiratory syncytial virus infection with sensitization by means of systemic (intraperitoneal) exposure with adjuvant. After sensitization, animals were similarly challenged with aerosolized allergen. Animals were treated with anti-IgE mAb to deplete IgE and to determine its role in the induction of allergic inflammation and mucosa pathology in these models. RESULTS: Concomitant respiratory syncytial virus infection significantly enhanced allergen sensitization by aerosol exposure and exacerbated eosinophilic inflammation and airway mucosa pathology. Depletion of IgE in this model significantly reduced lung eosinophilic inflammation and airway mucosa pathology. However, in the model in which animals were sensitized by means of systemic allergen exposure with adjuvant, depletion of IgE had no ameliorative effect on lung inflammation or pathology. CONCLUSION: We demonstrated that the method of antigen sensitization can delineate the role of IgE in allergen-induced lung inflammation. In a murine model that more closely resembles ambient allergen exposure in human subjects, IgE had a critical role in the pathogenesis of allergic asthma and mucosa pathology. The results parallel the results reported with anti-IgE efficacy in allergic asthmatic human subjects. PMID- 11398081 TI - Gene expression of vascular endothelial growth factor and its receptors and angiogenesis in bronchial asthma. AB - BACKGROUND: Angiogenesis is a feature of airway remodeling in bronchial asthma. The mechanism responsible for this angiogenesis is unknown. Vascular endothelial growth factor (VEGF) is a potent inducer of endothelial cells, which may contribute to chronic inflammation and angiogenesis. OBJECTIVE: We sought to investigate the molecular mechanisms underlying increased vascularity, and we examined the mRNA expression of VEGF and its receptors (flt-1 and flk-1) within bronchial biopsy specimens from asthmatic patients and normal control subjects. METHODS: Endobronchial biopsy specimens were examined immunocytochemically by staining with anti-type IV collagen mAb to evaluate vessel density by using computer-assisted image analysis. Specimens were also analyzed for the presence of the mRNAs of VEGF and its receptors with in situ hybridization. RESULTS: The extent of airway vascularity was increased in asthmatic subjects compared with that in control subjects (P <.01). Asthmatic subjects exhibited a greater expression of VEGF, flt-1, and flk-1 mRNA(+) cells in the airway mucosa compared with that in control subjects (P <.001 for each comparison). The degree of vascularity was associated with the number of VEGF, flt-1, and flk-1 mRNA(+) cells. Numbers of cells expressing VEGF mRNA inversely correlated with airway caliber (r = -0.83, P <.01) and airway hyperresponsiveness (r = -0.97, P <.001). Colocalization studies showed that macrophages, eosinophils, and CD34(+) cells were the major sources of VEGF; CD34(+) cells, macrophages, and T cells expressed both flt-1 and flk-1. CONCLUSION: These findings provide evidence that VEGF may play an important role in angiogenesis and subsequent airway remodeling in bronchial asthma. PMID- 11398082 TI - Inhibition of mast cell tryptase by inhaled APC 366 attenuates allergen-induced late-phase airway obstruction in asthma. AB - BACKGROUND: APC 366, a selective inhibitor of mast cell tryptase, has been shown to inhibit antigen-induced early asthmatic response (EAR), late asthmatic response (LAR), and bronchial hyperresponsiveness (BHR) in a sheep model of allergic asthma. OBJECTIVE: The purpose of this study was to investigate the effects of APC 366 on antigen-induced EAR, LAR, and BHR in mild atopic asthmatics not on any anti-inflammatory therapy. METHODS: Sixteen mild atopic asthmatics, each with a demonstrable antigen-induced EAR, LAR, and BHR to histamine, were recruited into this randomized, double-blinded, crossover study. APC 366 (5 mg)/placebo was administered by aerosol inhalation 3 times per day on treatment days 1 through 4. Allergen challenge was carried out on day 4. Histamine challenge was performed the following morning, 1 hour after final dosing. RESULTS: Subjects were shown to have a significantly smaller overall mean area under the curve for the LAR (P =.012) and mean maximum fall in FEV(1) for the LAR (P =.007) after pretreatment with APC 366 in comparison with placebo. No significant effects on BHR were demonstrable. Although the EAR was reduced by 18% after treatment with APC 366 in comparison with placebo, this was not statistically significant. CONCLUSION: Short-term repeated administration of APC 366 significantly reduced the magnitude of antigen-induced LAR in atopic asthmatics, which supports the role of mast cell tryptase in the pathophysiology of the LAR. PMID- 11398083 TI - Epidermal growth factor receptor signaling mediates regranulation of rat nasal goblet cells. AB - BACKGROUND: Mucus hypersecretion is a common response to inflammation in the lower airways and is a hallmark of chronic rhinitis. OBJECTIVE: The purpose of this study was to elucidate the mechanisms of regranulation (mucus production) of goblet cells in nasal epithelium. METHODS: Because neutrophils induce an epidermal growth factor (EGFR) cascade, we induced degranulation of goblet cells in rat nasal respiratory epithelium by means of intranasal inhalation of N-formyl methionyl-leucyl-phenylalanine (fMLP), and we examined regranulation of the goblet cells and the role of EGFR inhibitors and neutrophils in the regranulation process. RESULTS: In the control state Alcian blue/periodic acid-Schiff and mucin MUC5AC staining was present. Degranulation was induced in the nasal septal epithelium 4 hours after intranasal inhalation of fMLP (10(-7) mol/L); 48 hours later, goblet-cell regranulation was complete. In the control state EGFR protein staining was absent in the epithelium, but after fMLP-induced degranulation, EGFR protein was expressed. After pretreatment with BIBX1522, a selective EGFR tyrosine kinase inhibitor, fMLP-induced degranulation was unaffected, but goblet cell regranulation was prevented completely. CONCLUSION: These data suggest a role for the EGFR cascade in neutrophil-dependent production of goblet-cell mucins. Proving this theory will require the use of selective EGFR inhibitors in clinical studies of nasal hypersecretory states. PMID- 11398084 TI - Immunologic compensation in a patient with a large IgH constant region deletion. AB - BACKGROUND: Deficiencies of serum Ig of the IgG isotype typically predispose individuals to recurrent infections in some but not all cases. Patients with large deletions of the Ig heavy chain genes are free of recurrent and severe infections. OBJECTIVE: We sought to determine a mechanism of immunologic compensation that would possibly explain the reason for this patient's paucity of infection despite lacking several classes of serum Ig. METHODS: The patient is a 50-year-old white man. Serum Ig levels and specific antibody titers were measured by using various methods, including nephelometry, enzyme immunoassay, and radial immunodiffusion. The status of the Ig heavy chain genes was examined by means of Southern blotting of genomic DNA isolated from EBV-transformed B cells. RESULTS: The patient's serum lacked detectable IgG1, IgG2, IgG4, and IgA1 levels. Southern blot analysis demonstrated a large heavy chain constant (C) region gene deletion that included Cgamma1, Calpha1, psiCgamma, Cgamma2, and Cgamma4. Antibody responses to capsular pneumococcal and hemophilus polysaccharide antigens were essentially absent. However, IgG3 antibodies against the protein antigen tetanus toxoid were present. Relatively high antibody titers were found against pneumococcal surface proteins as well. CONCLUSION: We conclude that our patient's relative freedom from serious infection may be as a result of production of IgG3 antibodies to pneumococcal capsular proteins. PMID- 11398085 TI - Mechanisms of autoimmune activation of basophils in chronic urticaria. AB - BACKGROUND: Approximately 35% to 40% of patients with chronic urticaria possess a circulating antibody directed to the alpha subunit of the high-affinity type I IgE receptor (FcepsilonRI), which is detectable by using histamine release assays or immunoblotting. Prior reports suggest that purified IgG may not directly activate basophils but rather does so through complement activation. OBJECTIVE: We sought to further elucidate the mechanism by which this antibody causes basophil histamine release, including the role of complement, and to reassess the relationship of functional versus binding assays. METHODS: We incubated human basophils with patient serum, patient IgG, or patient IgG plus normal serum as a complement source and measured histamine release for each condition. IgG fractions were neutralized with cloned alpha subunit to determine whether histamine release decreased proportionately. We also screened sera from 260 patients to compare histamine release with immunoblotting results. RESULTS: We initially tested 35 sera from patients with chronic urticaria by using basophils from 2 atopic donors and one nonreleaser with rabbit anti-IgE. No histamine was released from the nonreleaser, yet all donors responded identically to monocyte chemotactic protein 1, indicating a requirement for IgE or the IgE receptor. Basophil histamine release was markedly augmented by complement if release by IgG alone was low. Incubation of purified IgG with an increasing concentration of cloned alpha subunit gradually reduced the histamine-releasing capability in patients with positive or negative immunoblot results. Of 260 patients tested, 43% had positive histamine release results, and 47% had positive immunoblot results, yet there was no correlation when individual patients were assessed. CONCLUSION: A subpopulation of patients with chronic urticaria possess IgG antibody directed to the alpha subunit of FcepsilonRI. This IgG activates basophils, which is dependent on or augmented by complement. Binding assays for the FcepsilonRI alpha subunit, such as immunoblotting, are not currently feasible as a screening method. A functional assay is required. PMID- 11398086 TI - Changes in sputum cell counts after exposure to occupational agents: what do they mean? AB - BACKGROUND: Exposure to occupational agents can induce eosinophilic inflammation in subjects with occupational asthma (OA). It might also induce nonspecific changes in airway inflammation in subjects without OA. OBJECTIVES: We sought to investigate the changes in airway inflammation induced by exposure to occupational agents in subjects with and without OA and to determine which changes in sputum eosinophil numbers and bronchial responsiveness to methacholine should be regarded as clinically significant for predicting a 20% fall in FEV(1). METHODS: We performed specific inhalation challenges (SICs) in 3 groups of subjects: subjects reporting a history consistent with OA with a positive SIC response (n = 17); subjects reporting a history consistent with OA with a negative SIC response (n = 14); and asthmatic subjects without any history of OA (n = 10). Induced sputum and methacholine challenges were performed at the end of the control day and again at the end of the last day of exposure; the last day of exposure was always performed in the laboratory. RESULTS: There was an increase in median sputum eosinophil and neutrophil numbers in subjects with positive SIC responses. Cell counts remained unchanged after exposure in asthmatic subjects without OA. A combination of a greater than 0.26 10(6)/mL increase in sputum eosinophil numbers and a decrease in the concentration of methacholine inducing a 20% fall in FEV(1) of at least 1.8-fold compared with baseline values predicted a 20% fall in FEV(1) in 96% (95% CI, 70%-99%) of patients. CONCLUSION: Exposure to occupational agents per se does not induce airway inflammation. Changes in both sputum eosinophil counts and methacholine responsiveness are satisfactory predictors of a significant bronchial responsiveness to occupational agents. PMID- 11398087 TI - Mutational analysis of the IgE epitopes in the latex allergen Hev b 5. AB - BACKGROUND: Hev b 5 is a major latex allergen and potential candidate for an immunotherapy reagent. OBJECTIVE: The purpose of this study was to produce a hypoallergenic form of Hev b 5. METHODS: We used SPOTs analysis with alanine substitution to identify amino acids (AAs) critical for IgE binding and used site directed mutagenesis to produce recombinant proteins with altered IgE-binding activity. RESULTS: Eleven epitopes were identified (5.1-5.11) in Hev b 5. Individual patients demonstrated variable epitope recognition, with the most intense reactivity to epitopes 5.4 and 5.7. IgE inhibition assays with synthetic peptides indicated that mutating a single epitope would not reduce IgE binding, but rather a combination of epitopes was required. After alanine substitutions to identify the important AAs, site-directed mutagenesis was used to replace the crucial AAs with alanine. Twenty clones with different combinations of altered epitopes were evaluated by means of IgE inhibition assays. Clones with mutations in single epitopes failed to reduce IgE binding, but changes to 8 epitopes (14 AAs) resulted in a 4500-fold reduction in IgE binding. Epitopes 5.7 and 5.9 were found to be cross-reactive, making Hev b 5 a multivalent allergen. CONCLUSIONS: We produced a recombinant Hev b 5 protein with significantly reduced IgE-binding activity. Changing a minimum of 3 immunodominant epitopes was required to cause a 100-fold reduction in IgE binding. Changes in 8 epitopes, particularly the cross reactive epitopes 5.7 and 5.9, were needed to maximize the reduction in IgE binding. Mutants with reduced IgE-binding activity may prove to be valuable reagents for immunotherapy. PMID- 11398088 TI - Effects of cooking methods on peanut allergenicity. AB - BACKGROUND: Allergy to peanut is a significant health problem. Interestingly, the prevalence of peanut allergy in China is much lower than that in the United States, despite a high rate of peanut consumption in China. In China, peanuts are commonly fried or boiled, whereas in the United States peanuts are typically dry roasted. OBJECTIVE: The aim of this study was to examine whether the method of preparing peanuts could be a factor in the disparity of allergy prevalence between the 2 countries. METHODS: Two varieties of peanuts grown in the United States were roasted, boiled, or fried. Proteins were analyzed by using SDS-PAGE and immunoblotting. Allergenicity was compared by using immunolabeling with sera from 8 patients with peanut allergy. RESULTS: The protein fractions of both varieties of peanuts were altered to a similar degree by frying or boiling. Compared with roasted peanuts, the relative amount of Ara h 1 was reduced in the fried and boiled preparations, resulting in a significant reduction of IgE binding intensity. In addition, there was significantly less IgE binding to Ara h 2 and Ara h 3 in fried and boiled peanuts compared with that in roasted peanuts, even though the protein amounts were similar in all 3 preparations. CONCLUSION: The methods of frying or boiling peanuts, as practiced in China, appear to reduce the allergenicity of peanuts compared with the method of dry roasting practiced widely in the United States. Roasting uses higher temperatures that apparently increase the allergenic property of peanut proteins and may help explain the difference in prevalence of peanut allergy observed in the 2 countries. PMID- 11398089 TI - Immunochemical characterization of edible bird's nest allergens. AB - BACKGROUND: We have previously described anaphylaxis induced by edible bird's nest (BN) and demonstrated that this condition is IgE mediated. OBJECTIVES: This study aimed at describing the immunochemical properties of the BN allergens. Comparative studies between 3 commercially available sources (according to the country of origin) of BN were also made. METHODS: Crude extracts of commercially available processed BN from Sarawak (Malaysia), Thailand, and Indonesia and fresh unprocessed BN from the caves of Sarawak were obtained by means of aqueous extraction. Specific IgE toward these sources were determined by using fluorescence allergosorbent tests (FASTs). Cross-reactivity studies between the 3 sources of commercially available processed BN were carried out by means of FAST inhibition. Immunochemical characterization by means of IgE immunoblot, periodate treatment, and heat stability studies were carried out on fresh unprocessed BN from Sarawak. RESULTS: Serum from allergic patients showed differences in IgE binding to the 3 sources of commercially available BN, with the highest levels of specific IgE recorded with the Sarawak source (P <.0001). Of these, only the Sarawak and Thailand sources showed considerable cross-reactivity. Further work on the unprocessed fresh Sarawak source identified a putative 66-kd major allergen containing several isoforms. Periodate treatment resulted in loss of IgE binding. Despite a progressive decline in the molecular weights of allergens on SDS-PAGE with increasing periods of boiling, IgE binding, as assessed by means of FAST, was not affected. N-terminal sequence of the major putative allergen (66 kd) showed homology to a domain of an ovoinhibitor precursor in chicken (SWISS PROT accession No. P10184). CONCLUSIONS: We have described the immunochemical properties of BN allergens. Edible BN from different sources are allergenically dissimilar. The putative major allergen is a 66-kd protein. PMID- 11398090 TI - Modification of T-cell receptor Vbeta repertoire in response to allergen stimulation in peanut allergy. AB - BACKGROUND: Peanut is one of the most common foods causing allergic reactions and is the most common cause of fatal and near-fatal food-related anaphylaxis. Little is known of the immunologic mechanisms that underlie peanut allergy. OBJECTIVES: In this study we examined clonality of the T-cell response (TCR) to peanut in MHC class II identical, peanut allergy-discordant sibling pairs. METHODS: Four sibling pairs were investigated. The TCR repertoire was analyzed before and after in vitro stimulation of PBMCs with crude peanut or PHA, as control for general/nonspecific reactivity. Eighteen TCR-Vbeta families were examined by flow cytometry. Where significant differences in incidence of particular TCR-Vbeta families were observed, PCR familyspecific cDNA amplification and gene scanning were performed. RESULTS: After stimulation with peanut, no selective expansion of any TCR-Vbeta subpopulation was observed with flow cytometry, in either the peanut-allergic or nonallergic siblings, with the exception of 1 peanut-allergic subject who demonstrated a significant increase of TCR-Vbeta11(+) cells (0.3% 5.9% of the total CD3(+) cells). However, gene scanning revealed predominant single-size PCR products for TCRBV11 in all peanut-allergic subjects after peanut stimulation. TCRBV11 polyclo-nality was observed in allergic and nonallergic subjects before peanut stimulation and in nonallergic subjects after peanut stimulation. In comparison, all subjects, before and after stimulation with peanut, showed polyclonality for TCRBV2. CONCLUSIONS: Our results argue for clonal or oligoclonal TCRs to crude peanut and indicate that changes in the TCRBV11 subpopulation are restricted to peanut-allergic subjects after stimulation with crude peanut allergen. PMID- 11398091 TI - Anaphylaxis caused by the new ant, Pachycondyla chinensis: demonstration of specific IgE and IgE-binding components. AB - BACKGROUND: There have been no reports dealing with the pathogenic mechanism and IgE-binding components in patients with anaphylaxis caused by a sting from Pachycondyla chinensis. OBJECTIVES: This study was conducted to observe the clinical features of patients with P chinensis -induced anaphylaxis. The roles of specific (s) IgE and sIgG4 antibodies were evaluated, and IgE-binding components were identified. METHODS: Seven patients with P chinensis -induced anaphylaxis and 15 unexposed control subjects were enrolled. P chinensis ants were collected at the patients' homes, and venom was prepared as P chinensis extract. Five patients complained of bee venom-induced anaphylaxis and had positive sIgE levels to yellow jacket venom, wasp venom, or both as well. Serum sIgE and sIgG4 were detected by means of ELISA. To identify IgE-binding components within P chinensis extracts, 12% SDS-PAGE with immunoblot analysis was applied. RESULTS: All patients had positive skin prick test responses to P chinensis antigen and positive sIgE levels. Five (71%) patients had positive sIgG4 levels. Eight IgE binding components (58, 46, 3l, 29, 27, 25, 22, and 12 kd) were noted, and the component at 12 kd was the most frequently found allergen (85%). IgE ELISA inhibition tests were performed on 2 groups of sera: one from patients with anaphylaxis induced by both P chinensis and bee venom (group A) and the other from patients with anaphylaxis induced by P chinensis venom alone without bee venom allergy (group B). ELISA inhibition tests with serum from group A showed significant inhibitions with addition of P chinensis extract, partial inhibitions with yellow jacket antigen, and minimal inhibitions with wasp or imported fire ant antigens. However, ELISA inhibition tests with serum from group B showed significant inhibitions with P chinensis antigen but no inhibition with wasp, yellow jacket, or imported fire ant antigens. CONCLUSIONS: IgE-mediated reactions contributed to the development of P chinensis -induced anaphylaxis. Eight IgE binding components and one major allergen (12 kd) were identified. Further studies will be needed to clarify the role of sIgG4 and to identify allergenic relationships with major bee and wasp allergens. PMID- 11398092 TI - The necessity for dual food intake to provoke food-dependent exercise-induced anaphylaxis (FEIAn): a case report of FEIAn with simultaneous intake of wheat and umeboshi. AB - BACKGROUND: Food-dependent exercised-induced anaphylaxis (FEIAn) is classified among the physical allergies. Many different food allergens have been reported, but the pathophysiology of FEIAn remains unknown. Furthermore, provocation tests with a suspected food do not always succeed in patients with FEIAn. OBJECTIVE: We sought to clarify and investigate causative foods and mechanisms of FEIAn in a 14 year-old boy. In addition, we tested in vivo and in vitro effects of cromolyn sodium in the same patient. METHODS: We used open challenge tests for the provocation of FEIAn and measured changes in plasma histamine levels and FEV1. In addition, we investigated the mechanism of FEIAn in this case with in vitro histamine release testing. RESULTS: The patient was diagnosed as having FEIAn by provocation testing with a simultaneous intake of wheat and umeboshi, but not when each food was eaten singly, followed by exercise. In addition, his plasma histamine level increased transiently and forced expiratory volume 1, expressed as a percentage change from baseline, decreased significantly. A synergistic effect on in vitro histamine release testing with 2 kinds of the causative foods was shown. Administration of cromolyn sodium proved to be effective on both the in vitro and in vivo tests. CONCLUSION: This is the first report of FEIAn provoked by the test with a simultaneous intake of 2 kinds of food. This case might in part explain negative challenge test results in patients with FEIAn. PMID- 11398093 TI - Vascular endothelial growth factor in patients with acute asthma. PMID- 11398094 TI - What is "more effective asthma control"? PMID- 11398098 TI - Optimal medical management of patients with chronic ischemic heart disease. PMID- 11398099 TI - Identification, by homozygosity mapping, of a novel locus for autosomal recessive congenital ichthyosis on chromosome 17p, and evidence for further genetic heterogeneity. AB - Autosomal recessive congenital ichthyosis (ARCI) comprises a group of severe disorders of keratinization, characterized by variable erythema and skin scaling. It is known for its high degree of genetic and clinical heterogeneity. Mutations in the gene for keratinocyte transglutaminase (TGM1) on chromosome 14q11 were shown in patients with ARCI, and a second locus was described, on chromosome 2q, in families from northern Africa. Three other loci for ARCI, on chromosomes 3p and 19p, were identified recently. We have embarked on a whole-genome scan for further loci for ARCI in four families from Germany, Turkey, and the United Arab Emirates. A novel ARCI locus was identified on chromosome 17p, between the markers at D17S938 and D17S1856, with a maximum LOD score of 3.38, at maximum recombination fraction 0.00, at D17S945, under heterogeneity. This locus is linked to the disease in the Turkish family and in the German family. Extensive genealogical studies revealed that the parents of the German patients with ARCI were eighth cousins. By homozygosity mapping, the localization of the gene could then be refined to the 8.4-cM interval between D17S938 and D17S1879. It could be shown, however, that ARCI in the two Arab families is linked neither to the new locus on chromosome 17p nor to one of the five loci known previously. Our findings give evidence of further genetic heterogeneity that is not linked to distinctive phenotypes. PMID- 11398100 TI - Linkage of otopalatodigital syndrome type 2 (OPD2) to distal Xq28: evidence for allelism with OPD1. AB - Otopalatodigital syndrome type 1 (OPD1) is an X-linked semidominant condition characterized by malformations of the skeleton, auditory apparatus, and palate. Previous studies have established linkage to a 16-cM region of Xq27-q28. A proposed allelic variant of OPD1, termed "OPD2," is associated with a more severe, frequently lethal phenotype with visceral and brain anomalies in addition to skeletal, auditory, and palatal defects. We report linkage of the OPD2 phenotype to a 2-cM region of distal Xq28 in a Maori kindred, with a maximum multipoint LOD score of 3.31 between the markers DXS1073 and DXS1108. This provides support for allelism between OPD1 and OPD2 and reduces the size of the disease interval to 1.8-2.1 Mb. We also demonstrate that female carriers of this disorder exhibit skewed inactivation that segregates with the high-risk haplotype and may be inversely related to the severity with which they manifest features of the disorder. PMID- 11398102 TI - Natural measles causes prolonged suppression of interleukin-12 production. AB - Among vaccine-preventable diseases, measles is the preeminent killer of children worldwide. Infection with measles virus (MV) is associated with prolonged suppression of cell-mediated immune responses, a phenomenon that is thought to underlie the susceptibility to secondary infections that accounts for most measles-related mortality. Interleukin (IL)-12 is critical for the orchestration of cellular immunity. MV specifically ablates IL-12 production by monocyte/macrophages in vitro through binding to CD46, a complement regulatory protein that is an MV receptor. To address the effect of MV on IL-12 responses in vivo, cytokine production was examined in Gambian patients with measles. IL-12 production by peripheral blood monocytes from such patients is markedly suppressed, which provides a unifying mechanism for many of the immunologic abnormalities associated with measles. This suppression is prolonged, with significant, stimulus-specific inhibition of IL-12 production demonstrable months after recovery from acute infection. However, despite this suppression, IL-12 responsiveness remains intact. PMID- 11398103 TI - A prospective case-control study of the role of astrovirus in acute diarrhea among hospitalized young children. AB - This study examines the importance of astroviruses as a cause of acute diarrhea in hospitalized children <10 years old during a 5-year period. Stools were screened by electron microscopy and were tested for astrovirus, rotavirus, and enteric adenovirus by EIA. During the study, 14.6% of hospitalized children had diarrhea. Astroviruses were second only to rotaviruses as etiologic agents of both community-acquired and nosocomial diarrhea. Community-acquired astrovirus infection occurred in 6.8% of patients, and nosocomial disease occurred in 16.2%. Most cases occurred from March through June, and astrovirus type 1 was the most common. The symptoms of astrovirus-infected children were similar to those of children with rotavirus infection. However, astrovirus-infected children had a lower median age, less dehydration, and lower symptom severity scores and were less likely to have been admitted for gastroenteritis than were children with rotavirus. Astrovirus, for which only rehydration therapy is required, should be considered as another common diarrheal pathogen in children <2 years old. PMID- 11398101 TI - Mutations of the protocadherin gene PCDH15 cause Usher syndrome type 1F. AB - Human chromosome 10q21-22 harbors USH1F in a region of conserved synteny to mouse chromosome 10. This region of mouse chromosome 10 contains Pcdh15, encoding a protocadherin gene that is mutated in ames waltzer and causes deafness and vestibular dysfunction. Here we report two mutations of protocadherin 15 (PCDH15) found in two families segregating Usher syndrome type 1F. A Northern blot probed with the PCDH15 cytoplasmic domain showed expression in the retina, consistent with its pathogenetic role in the retinitis pigmentosa associated with USH1F. PMID- 11398104 TI - Racial differences in HLA class II associations with hepatitis C virus outcomes. AB - A broad, vigorous CD4 T cell response, mediated by class II human leukocyte antigens (HLAs), favors hepatitis C virus (HCV) clearance. HLA-DQB1*0301 has been associated with viral clearance in an ethnically homogeneous cohort. To validate this association and to identify other class II associations in an ethnically varied cohort, molecular class II HLA typing was performed on 200 HCV clearance and 374 matched persistently infected subjects. HLA-DQB1*0301 was weakly associated with viral clearance in combined ethnic groups (odds ratio [OR], 0.72; 95% confidence interval [CI], 0.53-0.97) but was stronger in black subjects. In white subjects, viral clearance was associated with DRB1*0101 (OR, 0.32; 95% CI, 0.17-0.60) and its DQB1*0501 haplotype, whereas viral persistence was associated with DRB1*0301 (OR, 2.36; 95% CI, 1.23-4.52) and its DQB1*0201 haplotype. These results support a role for class II alleles in the immune response to HCV and underscore the importance of studying genetic associations in an ethnically diverse cohort. PMID- 11398105 TI - Human immunodeficiency virus type 1 stimulatory activity by Gardnerella vaginalis: relationship to biotypes and other pathogenic characteristics. AB - Stimulation of human immunodeficiency virus (HIV) type 1 expression by Gardnerella vaginalis is one possible cause for an increase in the amount of virus in the genital tract. The ability of G. vaginalis to induce HIV expression in chronically infected U1 cells was investigated, along with its possible relationship to biotype, genotype, and resistance to metronidazole and bacteriocin. Significant HIV stimulatory activity was found in 5 (50%) lysates of G. vaginalis. The ability to induce HIV expression in U1 cells was statistically associated with G. vaginalis biotype (P=.048) but not with genotype or resistance to metronidazole and bacteriocin. Further studies to explore the in vivo relevance of HIV activation by G. vaginalis in the female genital tract are warranted, since prevention strategies of bacterial vaginosis and colonization by certain biotypes of G. vaginalis may be valuable in reducing the risk of sexual transmission of HIV. PMID- 11398106 TI - Cellular replication of human immunodeficiency virus type 1 occurs in vaginal secretions. AB - Most human immunodeficiency virus type 1 (HIV-1) transmission worldwide is the result of exposure to infectious virus in genital secretions. However, current vaccine candidates are based on virus isolates from blood. In this study, vaginal secretions from HIV-1-infected women were examined for evidence of cellular viral replication that produced virus with properties different from that in blood. Multiply spliced HIV-1 messenger RNA, which is found only in cells replicating virus, was detected in all vaginal lavage samples tested. There was a strong correlation between the amounts of multiply spliced HIV-1 messenger RNA and of cell-free HIV-1 RNA in the lavage samples. In addition, significant genotypic differences were found in cell-free virus from matched blood plasma and vaginal secretions. Moreover, drug resistance-associated mutations appeared in plasma virus several months before appearing in vaginal virus. These findings indicate that cellular replication of HIV-1 occurs in vaginal secretions and can result in a virus population with important differences from that in blood. PMID- 11398107 TI - The steady-state pharmacokinetics of efavirenz and nevirapine when used in combination in human immunodeficiency virus type 1-infected persons. AB - The steady-state pharmacokinetics of efavirenz and nevirapine, when used in combination to treat human immunodeficiency virus type 1 (HIV-1)-infected subjects, were investigated. HIV-1-infected persons who had used efavirenz (600 mg once daily) for > or =2 weeks were eligible for study inclusion. The plasma pharmacokinetics of efavirenz were determined over 24 h. Subsequently, nevirapine (400 mg once daily) was added to the regimen. After 4 weeks, the pharmacokinetics of efavirenz and nevirapine were assessed over 24 h. The differences between the pharmacokinetic parameters of efavirenz with and without nevirapine were analyzed, and the pharmacokinetics of nevirapine were compared with those in historical control patients. The exposure to efavirenz when combined with nevirapine was significantly decreased by 22% (area under the plasma concentration vs. time curve), 36% (minimum plasma concentration), and 17% (maximum plasma concentration). Nevirapine pharmacokinetics appear to be unaffected by coadministration of efavirenz, compared with data from historical control patients. PMID- 11398108 TI - T cell receptor dynamism of mucosal and systemic CD4+ T cells in the course of an immune response to Escherichia coli heat-labile enterotoxin. AB - The changes in T cell receptor (TCR) Vbeta expression, use, and clonality in mice orally challenged with Escherichia coli heat-labile enterotoxin (LT) were assessed. Use of the TCR Vbeta family and clonality were significantly changed at the single-cell level. In Peyer's patches of treated mice, use of TCR Vbeta6, Vbeta8, and Vbeta14 increased in CD4(+)CD44(+) T cells, compared with use in nontreated mice. On the other hand, use of TCR Vbeta1 and Vbeta8 was enhanced in splenic CD4(+)CD44(+) T cells. Intraepithelial lymphocytes isolated from LT challenged mice showed expanded clonality (e.g., Vbeta1, Vbeta2, Vbeta9, and Vbeta18) and altered TCR Vbeta use (e.g., Vbeta15, Vbeta16, and Vbeta17). These findings reveal that oral administration of LT has distinct effects on mucosal versus systemic alphabeta T cells for induction of CD4(+) T cells with selected Vbeta use. This most likely reflects the function of LT as a mucosal modulator. PMID- 11398109 TI - Functional study of antibodies against a fibrogenin-binding protein in Staphylococcus epidermidis adherence to polyethylene catheters. AB - Staphylococcus epidermidis is an important pathogen in foreign body-associated infections. In a previous study, we showed that a surface-located fibrinogen binding protein, termed Fbe, from S. epidermidis mediated the bacterial adherence to fibrinogen-coated surfaces in vitro. In the present study, we demonstrate that antibodies against Fbe can block adherence of S. epidermidis to fibrinogen-coated catheters, subcutaneously implanted catheters from rats, and peripheral venous catheters from human patients. PMID- 11398110 TI - Structure and dissemination of a chromosomal insertion element encoding macrolide efflux in Streptococcus pneumoniae. AB - Macrolide resistance associated with macrolide efflux (mef) has rapidly increased in Streptococcus pneumoniae. We defined the genetic structure and dissemination of a novel mefE-containing chromosomal insertion element. The mefE gene was found on the 5' end of a 5.5- or 5.4-kb insertion designated as the macrolide efflux genetic assembly (mega), which is found in > or =4 distinct sites of the pneumococcal genome. The element was transformable and conferred macrolide resistance to susceptible S. pneumoniae. The first 2 open-reading frames (ORFs) of the element formed an operon composed of mefE and a predicted adenosine triphosphate-binding cassette homologous to msrA. Convergent to this efflux operon were 3 ORFs with homology to stress response genes of Tn5252. Mega was related to the recently described mefA-containing element Tn1207.1 but lacked the genes necessary for transposition and had unique termini and insertion sites. In metropolitan Atlanta, macrolide resistance due to mega rapidly increased in S. pneumoniae by clonal expansion and horizontally by transformation. PMID- 11398111 TI - Complex relationship between acquisition of beta-lactam resistance and loss of virulence in Streptococcus pneumoniae. AB - In a previous study of a murine peritonitis model, no Streptococcus pneumoniae strains were found that were both clinically penicillin resistant and virulent. This study assessed the relationship between acquired resistance and virulence in single- and double-isogenic penicillin-resistant (Peni-R) mutants obtained by transformation of a virulent penicillin-susceptible recipient strain with pbp2b and pbpX polymerase chain reaction fragments from a Peni-R donor strain. Sequence analysis results of the pbp2b and pbpX alleles from these strains were in keeping with acquired penicillin resistance. The virulence of these strains was significantly reduced, which shows a relationship between beta-lactam resistance and loss of virulence. The phenotype of the 23.2x mutant remained stable after in vivo passage, which suggests that the pbpX gene is involved in growth, whereas virulent revertants of the 23.2b and 23.2b.2x mutants had no change in MIC. Compensatory mutations are implicated in the revival of virulence. PMID- 11398112 TI - Evolution of immunologic responsiveness of persons living in an area of endemic bancroftian filariasis: a 17-year follow-up. AB - On an island in which bancroftian filariasis is endemic, 29 microfilaremic and 16 "endemic normal" (EN) subjects initially studied in 1974-1975 were reevaluated 17 years later. Eleven persons remained microfilaremic, whereas 18 had cleared both microfilaremia and antigenemia. Despite decreased infection on the island, antibody levels remained relatively constant for the subjects with persistent microfilaremia (Mf(+/+)), in contrast to sharp decreases for both EN subjects and subjects with cleared microfilaremia (Mf(+/-)). Although clinically indistinguishable from the EN subjects, the Mf(+/-) group had antibody levels (IgG, IgG4, and IgE) significantly lower than those of the EN subjects. Lymphocyte responses to parasite antigens were marginally greater in Mf(+/-) than in Mf(+/+) subjects, but both groups remained less cell responsive (as measured by proliferation, interleukin-5, interleukin-10, interferon-gamma, and granulocyte-macrophage colony-stimulating factor) than did the EN subjects. These findings suggest that, for microfilaremic persons, complete clearance of infection is not sufficient to restore "normal" immune responsiveness; filarial infection may induce very long-term deficits in the ability to respond to parasite antigens. PMID- 11398113 TI - Heterogeneity in diphtheria-tetanus-acellular pertussis vaccine-specific cellular immunity during infancy: relationship to variations in the kinetics of postnatal maturation of systemic th1 function. AB - Cellular immunity to vaccines is highly variable during infancy. This study addressed the hypothesis that these responses are governed by the pace of maturational changes in adaptive immune competence, in particular, cellular functions that underlie the postnatal transition from Th2 to Th1 "bias." Tetanus specific cytokine responses were tracked in peripheral blood mononuclear cells collected from infants at months 2, 4, 6, 12, and 18. These were compared with polyclonal responses. Results show that the Th2 component of the vaccine response develops rapidly and remains stable, unlike interferon (IFN)-gamma production, which also is initiated early but commonly declines after the final priming dose at 6 months. However, between 12 and 18 months, the IFN-gamma component of the vaccine-specific response has a spontaneous resurgence that coincides with a parallel increase in overall IFN-gamma production capacity. The Th2 component of vaccine-specific responses was more prominent in children with atopic family history. PMID- 11398114 TI - CCR5 promoter polymorphisms in a Kenyan perinatal human immunodeficiency virus type 1 cohort: association with increased 2-year maternal mortality. AB - The CCR5 chemokine receptor acts as a coreceptor with CD4 to permit infection by primary macrophage-tropic human immunodeficiency virus type 1 (HIV-1) strains. The CCR5Delta32 mutation, which is associated with resistance to infection in homozygous individuals and delayed disease progression in heterozygous individuals, is rare in Africa, where the HIV-1 epidemic is growing rapidly. Several polymorphisms in the promoter region of CCR5 have been identified, the clinical and functional relevance of which remain poorly defined. We evaluated the effect of 4 CCR5 promoter mutations on systemic and mucosal HIV-1 replication, disease progression, and perinatal transmission in a cohort of 276 HIV-1-seropositive women in Nairobi, Kenya. Mutations at positions 59353, 59402, and 59029 were not associated with effects on mortality, virus load, genital shedding, or transmission in this cohort. However, women with the 59356 C/T genotype had a 3.1-fold increased risk of death during the 2-year follow-up period (95% confidence interval [CI], 1.0-9.5) and a significant increase in vaginal shedding of HIV-1-infected cells (odds ratio, 2.1; 95% CI, 1.0-4.3), compared with women with the 59356 C/C genotype. PMID- 11398115 TI - Cd4+ T cells programmed to traffic to lymph nodes account for increases in numbers of cd4+ T cells up to 1 year after the initiation of highly active antiretroviral therapy for human immunodeficiency virus type 1 infection. AB - Cells programmed to traffic through lymph nodes dominate initial increases in total CD4(+) T cell numbers after highly active antiretroviral therapy (HAART) is begun for human immunodeficiency virus type 1 (HIV-1) infection. However, it is unknown whether this dominance continues throughout the first year of treatment. To examine this question, 10 subjects who had a positive response to HAART for 1 year were selected from a cohort of 20 who were receiving this treatment. Flow cytometry, which was used to characterize CD4(+) T cell subsets by immunophenotype, demonstrated that cells programmed to traffic through lymph nodes, irrespective of their memory or naive phenotype, continued to best account for increases in CD4(+) T cells, even 1 year after starting HAART. This suggests that, although this pool is preferentially depleted during HIV-1 infection, HAART allows for reaccumulation of these cells for at least 1 year. Furthermore, it suggests that phenotypic differences based on markers of lymphocyte trafficking may be more relevant for understanding HIV-1 pathogenesis than are naive and memory markers alone. PMID- 11398116 TI - Prevention of meningococcal serogroup B infections in children: a protein-based vaccine induces immunologic memory. AB - Immunologic memory against meningococci was studied in 177 children (100 children were 10-11 years old and 77 were 5-6 years old) 2.5 years after vaccination with hexavalent meningococcal outer membrane vesicle (OMV) vaccine or hepatitis B (HepB) vaccine. Children were revaccinated with monovalent P1.7(h),4 meningococcal OMV vaccine. Serum bactericidal antibodies (SBAs) were measured before revaccination and after 4-6 weeks. A minimum 4-fold increase in SBAs against serosubtype P1.7(h),4 was detected in 48.5% of the children after hexavalent meningococcal vaccine and in 8.9% after HepB vaccine. Of the initial responders given hexavalent meningococcal vaccine, 78% had > or =4-fold increase in SBAs against strain P1.4. Thus, immunologic memory is present in toddlers and school-aged children previously given 3 hexavalent meningococcal vaccinations. Booster vaccination with monovalent P1.7(h),4 meningococcal OMV vaccine induces a significant increase in SBAs against serosubtype P1.7(h),4 and cross-reactivity against other serosubtypes in the hexavalent vaccine. PMID- 11398117 TI - Treatment of diarrhea caused by Cryptosporidium parvum: a prospective randomized, double-blind, placebo-controlled study of Nitazoxanide. AB - A prospective randomized, double-blind, placebo-controlled study was conducted in 50 adults and 50 children from the Nile delta of Egypt, to evaluate the efficacy of nitazoxanide in treating diarrhea caused by Cryptosporidium parvum. Nitazoxanide was administered in 500-mg doses twice daily for 3 days in adults and adolescents, in 200-mg doses twice daily for 3 days in children aged 4-11 years, and in 100-mg doses twice daily for 3 days in children aged 1-3 years. At 7 days after initiation of therapy, diarrhea had resolved in 39 (80%) of the 49 patients in the nitazoxanide treatment group, compared with 20 (41%) of 49 in the placebo group (P<.0001). Diarrhea was resolved in most patients receiving nitazoxanide within 3 or 4 days of treatment initiation. Nitazoxanide treatment reduced the duration of both diarrhea (P<.0001) and oocyst shedding (P<.0001). PMID- 11398118 TI - Fcgamma receptor IIa (CD32) polymorphism is associated with protection of infants against high-density Plasmodium falciparum infection. VII. Asembo Bay Cohort Project. AB - In vitro studies have shown that inhibition of Plasmodium falciparum blood-stage parasite growth by antibody-dependent cellular inhibition is mediated by cooperation between malaria-specific IgG1 and IgG3, but not IgG2, and monocytes via the Fcgamma receptor II (FcgammaRII). A single amino acid substitution at position 131 in FcgammaRIIa is critical in the binding of human IgG subclasses. The hypothesis that the FcgammaRIIa-Arg/Arg131 genotype, which does not bind to IgG2, is a host genetic factor for protection against high-density P. falciparum infection was tested. One hundred eighty-two infants from a large community-based birth cohort study in western Kenya were selected for an unmatched case-control study. Results showed that the infants with the FcgammaRIIa-Arg/Arg131 genotype were significantly less likely to be at risk for high-density falciparum infection, compared with infants with the FcgammaRIIa-His/Arg131 genotype (adjusted odds ratio, 0.278; 95% confidence interval, 0.123-0.627; P=.0021). This finding supports the hypothesis. PMID- 11398119 TI - Distinct immunity in patients with visceral leishmaniasis from that in subclinically infected and drug-cured people: implications for the mechanism underlying drug cure. AB - Significant levels of IgG3 and IgG4 and high levels of IgG1 leishmania-specific antibody differentiated the immune states in 10 patients with visceral leishmaniasis from those of virtually all 20 drug-cured and 18 subclinically infected subjects, whereas the level of IgG2 antibody was nondiscriminating. The most extreme "subclinically infected" outlier subsequently developed disease. Overall, the immune states in subclinically infected and drug-cured persons were mutually indistinguishable but were readily distinguished from those of patients. These findings may have implications for the immunologic mechanism underlying drug cure in visceral leishmaniasis. PMID- 11398120 TI - Meningococcal C polysaccharide vaccine induces immunologic hyporesponsiveness in adults: additional data. PMID- 11398122 TI - Granulocyte transfusion therapy: donor-related risks. PMID- 11398124 TI - Suction injuries: education is the key to prevention. PMID- 11398125 TI - Adopting the qualitative paradigm to understanding children's perspectives of illness: barrier or facilitator? AB - Reliance on the traditional quantitative paradigm has been the norm for exploring the phenomenon of children's illness experiences. Although it provides valuable information on the psychosocial sequelae of children in response to illness, an account of children's perspectives of the illness experience is missing. Accordingly, researchers since the early 1980s have been advocating the need to adopt the qualitative paradigm in the study of children's illness experiences. Nonetheless, even though there is a call for more qualitative research, the majority of research being conducted today still supports the quantitative paradigm. This report examines why researchers may be reluctant to adopt the qualitative paradigm. Specifically, the author proposes that this reluctance may stem from researchers' perceiving children as being less competent and social than adults, which would explain the need to research children solely from an objective, impersonal stance. Primary philosophical principles of the qualitative paradigm may be seen as barriers to accessing children's thoughts. The author suggests, however, that when children are viewed from an alternative perspective, the major principles of the qualitative paradigm may actually serve as facilitators to apprehending their thoughts and feelings. The following key principles of the qualitative research paradigm are examined: (a) realities are constructed by human beings who are viewed as active agents making sense out of the realities they encounter; (b) realities are multiple and must be perceived holistically and from various vantage points; and (c) realities are shared and mutually shaped by the researcher and research participants. Key strategies that promote successful use of the qualitative paradigm in the study of children's illness experiences are also discussed. PMID- 11398126 TI - Biobehavioral characteristics of infants with failure to thrive. AB - Failure to thrive (FTT) is a syndrome of growth failure that results in an infant who is behaviorally difficult. The current thinking is that FTT results from a problematic infant-mother interaction, with the infant making a significant contribution to the interactional process. It is possible that the behavioral characteristics of the infant with FTT may be related to underlying physiologic response patterns, specifically, activity of the autonomic nervous system. The purpose of this study is to examine the relationships among behavioral responsiveness, heart rate variability as a marker of autonomic nervous system activity, and nutritional status in infants with FTT. Infants with FTT were matched with healthy growing infants (n = 14 pairs). Results from the study indicated that infants with FTT exhibited considerably more negative behaviors and exhibited low heart rate variability. It appears that there may be a physiologic basis to the behaviors that are exhibited by infants with FTT. Prospective research is needed to further clarify this relationship. PMID- 11398127 TI - A cross-cultural comparison of resilience in adolescents. AB - The purpose of this study was to continue the process of clarifying the concept of resilience in adolescence. At the completion of the first phase of this clarification process in 1997, it became evident that adolescents believed resilience hurt them more than helped them and encompassed such dimensions as self-protection and survival. To gain the broadest understanding of the adolescent's perception of resilience, this study qualitatively explored those perceptions from adolescents in varied socioeconomic and cultural environments. Using a focus-group format, we queried 40 adolescents from New England and Ghana about their perceptions of adversity, overcoming adversities, and resilience. The results indicated that irrespective of age, gender, culture, and socioeconomic status, all the adolescents believed that they were resilient; however, overcoming adversities and being resilient were different depending on the presence or absence of consistent, loving, caring, mentoring adults who helped the adolescent traverse the adversities of life. Those adolescents who were without such support systems (found predominately in the New England sample) showed survival and self-protective forms of resilience, whereas those with such support systems (found predominately in the Ghanaian sample) showed a connected form of resilience. Further research is needed; however, adolescents have given clear messages in two studies indicating that being resilient can hurt them as much as it may help them. When fostering resilience, consideration should be given to what kind of resilience is being fostered. PMID- 11398128 TI - Using Touchpoints to promote parental self-competence in low-income, minority, pregnant, and parenting teen mothers. AB - This pilot study was designed to test an intervention developed to increase parenting self-confidence in teen mothers. Group discussions of personal experience and parenting were substituted for a traditional child development course. Using a Touchpoints approach, teachers focused on strengths of the adolescents and their children. Information about child development was presented as it naturally occurred. A total of 20 students took part in the study. There was a significant increase in parenting self-confidence after the intervention. Although the sample size was small, the results were encouraging with this group of teens. A larger study is planned to continue development of the intervention. PMID- 11398129 TI - Adolescent cigarette smoking and health risk behavior. AB - During the past 30 years, tobacco use among adolescents has substantially increased, resulting in major health problems associated with tobacco consumption. The purpose of this study was to identify adolescent smoking behaviors and to determine the relationship among smoking, specific demographic variables, and health risk behaviors. The sample consisted of 93 self-selecting adolescents. An ex post facto design was used for this study and data were analyzed by using nonparametric statistics. Findings included a statistically significant relationship between lifetime cigarette use and ethnicity. Statistically significant relationships were also found among current cigarette use and ethnicity, alcohol use, marijuana use, suicidal thoughts, and age at first sexual intercourse. Nurses and other providers must recognize that cigarette smoking may indicate other risk behaviors common among adolescents. PMID- 11398130 TI - Adolescent school health. PMID- 11398131 TI - Children and pets. PMID- 11398132 TI - Preparing the children. PMID- 11398133 TI - Qualitative analysis of the care of children in hospital in four countries-Part 2. AB - As part of a large study of the care of children in Australian, British, Indonesian, and Thai hospitals, qualitative methods were used to examine differences influenced by culture. Two groups were surveyed: parents of hospitalized children, and staff caring for them. Vignettes were used to invoke discussion, and content analysis was used to examine the data. Subjects were interviewed singly, or in focus groups. These interviews were audiotaped and transcribed for analysis. This article is the second of a two-part series, and includes results of the staff's interviews and discussion. The parents' results and discussion were published in Part 1 (Shields & King, 2001). Staff in all the countries considered communication with parents to be an important part of care of the hospitalized child, and this was consistent with the parents' responses. Staff were mindful of safe practices, though more so in Australia and Britain than Indonesia and Thailand. Cost of treatment for the parents was an important consideration for staff in Indonesia and Thailand when they were planning care for the child. Cultural constructions were more likely to be considered by the Australian and British staff than the Indonesian and Thai staff, and this may have been influenced by the prevailing culture of medical dominance in those countries. PMID- 11398134 TI - Predictors for development of hyperuricemia: an 8-year longitudinal study in middle-aged Japanese men. AB - To identify the factors responsible for increases in serum uric acid (SUA), a cohort of 1,312 hyperuricemia-free (SUA < 7.5 mg/dL and no medication for hyperuricemia or hypertension) male office workers aged 30 to 52 years were examined annually for 8 successive years. Subjects who were found to have become hyperuricemic (SUA > or = 7.5 mg/dL) or who started medication for hyperuricemia during repeat surveys were defined as incidence cases. The SUA trend was also examined in 1,062 subjects for whom 9 consecutive SUA values were available and who did not start medication for hyperuricemia or hypertension during the observation period. Multivariate analyses, excluding the baseline SUA level as a factor in the Cox proportional-hazards model, indicated that age (negative), body mass index (BMI), log triglyceride level, hemoglobin A(1c) (HbA(1c)) level (negative), white blood cell count, and alcohol intake at study entry were significantly associated with the incidence of hyperuricemia. In the model including the baseline SUA level, baseline SUA level was the strongest factor for the incidence of hyperuricemia, and BMI, white blood cell count, and alcohol intake at study entry remained as independent factors. From stepwise linear regression analyses for SUA slope, excluding the baseline SUA level as a factor, significant correlates with SUA slope were, in order of their relative importance, slopes of BMI, HbA(1c) (negative), blood urea nitrogen, log triglyceride level, total protein, and baseline levels of hematocrit (negative), white blood cells, and HbA(1c) (negative). In stepwise linear regression analyses, including the baseline SUA level as a factor, SUA level (negative) and alcohol intake at study entry emerged as significant factors for SUA slope. The cumulative percentage of variation for SUA slope was 25.6%. In conclusion, obesity, alcohol intake, and multimetabolic disorders were determined to be independent predictors for the development of hyperuricemia. In addition, the white blood cell level may be a contributory factor. PMID- 11398135 TI - Changes in serum hypoxanthine levels by exercise in obese subjects. AB - To study on effect of obesity on changes in serum hypoxanthine with exercise, exercise stress testing with treadmill was performed on 7 obese subjects (body mass index [BMI], 30.6 +/- 3.2 kg/m(2)) and 16 healthy volunteers (BMI, 21.5 +/- 2.10 kg/m(2)). Expiratory gas analysis during exercise showed that peak Vo(2) was significantly lower in the obese group than in the control group (28.1 +/- 4.0 v 37.1 +/- 4.7 mL/kg/min; P <.001). Furthermore, the obese group had lower anaerobic threshold (AT) values (P <.005), respiratory quotient at AT (P =.003), and exercise capacity reserve (P =.002) than the control group. Baseline serum hypoxanthine levels were significantly higher in the obese group than in the control group (3.46 +/- 3.70 v 1.23 +/- 1.16 micromol/L; P <.05). Exercise induced a pronounced increase in serum hypoxanthine level in the obese group compared with the control group (10.65 +/- 6.81 v 43.86 +/- 4.56 micromol/L; P <.01). Serum levels of uric acid before and after load were also higher in the obese group than in the control group (404 +/- 43 v 302 +/- 77 micromol/L; P <.005). A pronounced increase in hypoxanthine with exercise may result in organ damage caused by free radicals, and intermittent training from mild intensity may be less hazardous for exercise treatment of obesity. PMID- 11398136 TI - Polymorphisms of the insulin gene among Japanese subjects. AB - We have sequenced the insulin gene in 72 unrelated Japanese subjects (52 with type 2 diabetes mellitus and 20 with normal glucose tolerance). We identified 6 mutations and all were found at a low frequency (1% to 4%). Three mutations were new. These included a C-to-G substitution in the promoter region, a G-to-A substitution in codon-2 resulting in an Ala-to-Thr replacement in amino acid -2 of the signal peptide, and a G-to-A substitution in intron 2. We have no evidence that any of the mutations that we found are the cause of diabetes. Thus, mutations in the insulin gene do not appear to be an important genetic factor contributing to the development of diabetes in this population. PMID- 11398138 TI - Factors explaining the difference of total homocysteine between men and women in the European Investigation Into Cancer and Nutrition Potsdam study. AB - Interestingly, plasma total homocysteine (tHcy) concentration is consistently higher in men than in women. This observation deserves further investigations because elevated tHcy concentrations have been shown to be independently associated with coronary, peripheral, and cerebral vascular diseases. It was the aim of the present study to define major determinants of plasma tHcy in a healthy middle-aged German population under particular consideration of the gender factor. The study population was obtained from an ongoing recruitment procedure for a cohort study and comprised 336 men and women, aged 40 to 65 years. Exclusion criteria were elevated creatinine levels in blood, history of skin or atherosclerotic diseases, current use of vitamins or other supplements, and heavy smoking. Plasma tHcy, folate, vitamin B12, vitamin B6, creatinine, testosterone and estradiol, protein, and hematocrit were measured. Fat-free mass was assessed by skinfold thickness. The C677T polymorphism of the methylenetetrahydrofolate reductase (MTHFR), a key enzyme of folate and homocysteine metabolism, was determined by polymerase chain reaction (PCR) with restriction enzyme analysis. In this population, plasma tHcy ranged from 5 to 46 micromol/L. The frequency of the T allele of the MTHFR was 0.29, which is lower than in other populations. A total of 54.2% of this population was homozygote for the wild-type, 39.6% heterozygote, and 6.2% homozygote for the mutation. tHcy correlated negatively with folate and cobalamin concentration in blood and positively with creatinine. No correlation was seen with vitamin B6. From the gender-related variables, tHyc correlated significantly with fat-free mass and testosterone and inversely with estradiol. The difference between gender with regard to tHcy was mainly explained by differences in fat-free mass, but also by estradiol concentrations. The following contributions to the variation of tHcy were seen in a multivariate regression model: plasma cobalamin (11%), creatinine (11%), plasma folate (8%), fat-free mass (5%), estradiol (2%), MTHFR polymorphisms (2%), and plasma protein (1%). We concluded that tHcy in the general population has a variety of determinants ranging from nutrition, internal metabolic parameters to gender related variables. PMID- 11398137 TI - Effect of low-density lipoproteins on apolipoprotein AI kinetics in heterozygous familial hypercholesterolemia. AB - In patients with heterozygous familial hypercholesterolemia (FH), both synthetic and clearance rates of high-density lipoproteins (HDL) are increased compared with control subjects. According to in vitro data on hepatocytes, the expanded pool size of low-density lipoproteins (LDL) in FH could partly explain the enhanced HDL production. Therefore, we have tested the hypothesis that a reduction of LDL pool size, achieved by LDL-apheresis, is associated with a downregulation of HDL synthesis. We studied the kinetics of HDL by infusing [5,5,5-(2)H(3)]-leucine in 7 heterozygous FH patients before and after 3 biweekly LDL-apheresis using dextran sulfate columns. Both plasma and LDL-cholesterol levels were decreased after LDL-apheresis (169 +/- 35 v 422 +/- 27 mg/dL, P <.05, and 85 +/- 19 v 327 +/- 52 mg/dL, P <.05, respectively). Plasma triglyceride level was unaffected (162 +/- 43 v 176 +/- 35 mg/dL, not significant [NS]) and HDL composition remained stable (HDL-cholesterol 29 +/- 6 v 37 +/- 7 mg/dL, NS, and HDL-triglyceride 20 +/- 6 v 19 +/- 8 mg/dL, NS). Plasma apolipoprotein AI (apo AI) was also similar (122 +/- 20 v 115 +/- 18 mg/dL, NS). Mean HDL-apo AI fractional catabolic rate (FCR) was slightly higher (0.41 +/- 0.07 v 0.36 +/- 0.14 pool/d, NS), and absolute production rate (APR) was increased (22.1 +/- 5.7 v 18.0 +/- 5.7 mg/kg/d, P <.05) after LDL-apheresis. These human kinetic data suggest that LDL do not play a major role on HDL production in heterozygous FH patients. PMID- 11398139 TI - Altered purine nucleotide degradation during exercise in patients with essential hypertension. AB - Purine degradation occurs during strenuous muscle exercise and plasma levels of hypoxanthine (HX), purine degradation intermediate, increase. Purine nucleotide degradation has not been investigated in patients with essential hypertension (HTN). The present study determined whether purine nucleotide degradation is altered in patients with HTN. Cardiopulmonary exercise test was performed with serial measurements in blood lactate and plasma HX in 24 patients (14 men and 10 women) with essential HTN (World Health Organization [WHO] class I to II; mean age, 57.7 +/- 2.1 years) and 24 age-, sex-matched normal subjects. Exercise was terminated either by severe fatigue or excess blood pressure increase. Peak work rate (WR) (normal v HTN, 151 +/- 10 v 135 +/- 8 W, not significant [NS]) was not different, but peak oxygen uptake (peak Vo(2), 26.3 +/- 1.5 v 22.2 +/- 0.9 mL/min/kg, P <.05) and anaerobic threshold were lower in patients with HTN. Resting levels of blood lactate and plasma HX were similar, but the increment from rest to peak exercise (Delta) for lactate (Delta lactate: 4.4 +/- 0.4 v 3.4 +/- 0.4 mmol/L, P <.05) and for HX (Delta HX, 15.9 +/- 2.2 v 9.1 +/- 1.1 micromol/L, P <.05) were significantly smaller in patients with HTN. When normalized by the peak WR, Delta HX/peak WR (0.105 +/- 0.013 v 0.069 +/- 0.007 micromol/L/W, P <.05) was significantly lower in patients with HTN. Patients with HTN exhibited reduced HX response to exercise with impaired exercise capacity. The exercise-induced changes in plasma HX were smaller in patients with HT when normalized with peak WR. These results suggest that the purine nucleotide degradation is reduced in patients with HTN. PMID- 11398140 TI - Association between the TaqIB polymorphism in the cholesteryl ester transfer protein gene locus and plasma lipoprotein levels in familial hypercholesterolemia. AB - Cholesteryl ester transfer protein (CETP) facilitates the exchange of triglycerides (TG) and cholesteryl ester between lipoprotein particles. Subjects with familial hypercholesterolemia (FH) have been reported to have higher CETP activities, which could contribute to the lower high-density lipoprotein cholesterol (HDL-C) levels and increased cardiovascular risk observed in some of these patients. Several polymorphisms have been reported in the CETP locus; the common TaqlB polymorphism is associated, in normolipidemic subjects, with decreased CETP activity and levels and with increased HDL-C levels. No data is available on the influence of this polymorphism in FH subjects. We have examined the TaqIB polymorphism in a group of 101 FH heterozygotes from Valencia, Spain. We have observed a frequency of 0.43 for the B2 allele, similar to those reported in the general population. Based on analysis of variance (ANOVA), we found significant associations between the presence of the B2 allele and increased plasma HDL-C (P <.04) and apolipoprotein A-I (apoA-I) levels (P <.01). An opposite association was observed for low-density lipoprotein-cholesterol (LDL-C) levels, with the B2/B2 subjects having lower levels than B1/B1 and B1/B2 subjects. The plasma apoB levels followed the same trend as those for LDL-C. In addition, the response to a National Cholesterol Education Program (NCEP)-I diet was studied in 77 of these subjects. The TaqlB polymorphism did not have a significant effect over the individual dietary response for any of the variables examined, as demonstrated by the lack of significant gene by diet interactions. In summary, the CETP TaqlB polymorphism is associated with a less atherogenic lipid profile, consisting of lower LDL-C, higher HDL-C levels, and a lower LDL C/HDL-C ratio in heterozygous FH subjects. Moreover, the B2 allele was associated with a lower appearance of arcus cornealis, xanthomata, and clinical arteriosclerotic disease in these subjects. PMID- 11398141 TI - Modulation by blood glucose levels of activity and concentration of paraoxonase in young patients with type 1 diabetes mellitus. AB - Paraoxonase (PON) is a high-density lipoprotein (HDL)-associated esterase, which may prevent the transformation of low-density lipoproteins (LDL) into biologically active, atherogenic particles. PON concentration and activity are affected by PON1 gene polymorphisms and found to be altered in type 2 diabetes patients with retinopathy. We investigated serum PON concentration, in vitro activity and polymorphism at position 54 (L/M, Leu-Met54) in 193 Caucasian adolescents and young adults (88 males, 105 females) with type 1 diabetes mellitus, as well as its relationship to the presence of retinopathy. An inverse linear correlation was found between blood glucose levels and both serum PON concentration (r = -.20, P =.017) and its activity (r = -0.17, P =.037). Patients with elevated blood glucose values (> or =10 mmol/L) had significantly lower levels of both PON concentration (P =.003) and activity (P =.028) than those with lower glucose levels. After adjusting for blood glucose and diabetes duration, PON activity was significantly higher in patients with different stages of retinopathy compared with those without retinopathy (P =.003). The L/L genotype was closely associated with the presence of retinopathy (P <.0001). These data show that young people with type 1 diabetes and the L/L polymorphism at position 54 of PON1 gene are more susceptible to retinal complications. However, the role of serum PON concentration and activity as a possible marker for monitoring late microvascular complications in these patients has to be established. PMID- 11398142 TI - Angiotensin regulates endothelin-B receptor in rat inner medullary collecting duct. AB - Our recent studies showed that endothelin (ET)(B) receptors are downregulated in congestive heart failure. These changes in ET(B) receptor density can be prevented by angiotensin-converting enzyme inhibitors, suggesting a possible role for angiotensin. Using isolated inner medullary collecting ducts (IMCD), we examined the possibility that angiotensin-induced downregulation of ET(B) receptors is accompanied by a decrease in ET(B) receptor mRNA. Binding studies showed that overnight incubation with angiotensin II induced a downregulatiion of ET(A) and ET(B) receptors' density in IMCD by 39% and 29%, respectively. This downregulation in ET receptor density was abolished when IMCD was coincubated with angiotensin II and its receptor antagonist saralasin. Furthermore, when the cells were exposed to phorbol myristate acetate (PMA), it resulted in a reduction in ET(A) and ET(B) receptor binding sites by 41% and 34%, respectively, suggesting the involvement of protein kinase C (PKC). In isolated IMCD, ET-1 induced an increase in cyclic guanosine monophosphate (cGMP) accumulation (705 + 63 to 1,015 + 88 fmol/microg protein/5min, P <.01), and the ET-1-induced accumulation was attenuated in the presence of angiotensin II (641 + 45 to 809 + 46 fmol/microg protein/5min, P <.01). Using competitive polymerase chain reaction (PCR) method, we also observed downregulation of ET(A) and ET(B) receptors mRNA in IMCD treated with angiotensin II (ET(A), 1.09+0.11 v 0.77 + 0.07 amol/microg of total RNA, P <.01; ET(B), 14.80 + 1.95 v 8.65 + 0.67 amol/microg of total RNA, P <.01). The addition of a PKC inhibitor abolished the downregulation of ET(A) and ET(B) receptor mRNA induced by angiotensin II (ET(A), 1.25 + 0.07 v 1.19 + 0.06 amol/microg of total RNA, not significant [NS]; ET(B), 14.36 + 0.83 to 13.68 + 0.64 amol/microg of total RNA, NS). These results suggest that angiotensin II induced downregulation of ET(A) and ET(B) receptors mRNA is mediated by a mechanism involving PKC. PMID- 11398143 TI - Effect of vanadium on insulin sensitivity and appetite. AB - Vanadium, a potent nonselective inhibitor of protein tyrosine phosphatases, has been shown to mimic many of the metabolic actions of insulin both in vivo and in vitro. The mechanism(s) of the effect of vanadium on the decrease in appetite and body weight in Zucker fa/fa rats, an insulin-resistant model, is still unclear. Because insulin may inhibit hypothalamic neuropeptide Y (NPY), which is known to be related to appetite, and increase leptin secretion in adipose tissue, we studied the possibility that the changes in appetite produced by vanadium may be linked to altered NPY levels in the hypothalamus. We also examined effects of vanadium on leptin. Zucker lean and fatty rats were chronically treated with bis(maltolato)oxovanadium(IV) (BMOV), an organic vanadium compound, in the drinking water. Plasma and adipose tissue leptin levels were measured by radioimmunoassay and immunoblotting, respectively. Hypothalamic NPY mRNA and peptide levels were measured using in situ hybridization and immunocytochemistry, respectively. BMOV treatment significantly reduced food intake, body fat, body weight, plasma insulin levels, and glucose levels in fatty Zucker rats. Fifteen minutes after insulin injection (5 U/kg, intravenous [IV]), circulating leptin levels (+100%) and adipose leptin levels (+60%) were elevated in BMOV-treated fatty rats, although these effects were not observed in untreated fatty rats. NPY mRNA levels in the arcuate nucleus (ARC) (-29%), NPY peptide levels in ARC ( 31%), as well as in the paraventricular nucleus (PVN) (-37%) were decreased with BMOV treatment in these fatty rats. These data indicate that BMOV may increase insulin sensitivity in adipose tissue and decrease appetite and body fat by decreasing NPY levels in the hypothalamus. BMOV-induced reduction in appetite and weight gain along with normalized insulin levels in models of obesity, suggest its possible use as a therapeutic agent in obesity. PMID- 11398144 TI - Lack of in vivo effect of vanadium on GLUT4 translocation in white adipose tissue of streptozotocin-diabetic rats. AB - Vanadium treatment, in vivo, corrects the severe hyperglycemia observed in streptozotocin (STZ)-diabetic rats. A number of metabolic effects of vanadium have been demonstrated in vitro and might contribute importantly to normalization of glucose homeostasis. However, many in vitro effects of vanadium occur at concentrations substantially higher than those achieved in vivo. Effects of vanadium on white adipose tissue have been particularly well characterized in vitro. To examine the relationship between in vitro and in vivo actions of vanadium, we examined the effects of vanadium treatment on acute glucose tolerance and adipose tissue GLUT4 control in vivo. In agreement with previous studies, vanadium treatment of STZ-diabetic rats restored normoglycemia with no appreciable restoration of insulin secretion. GLUT4 expression in white adipose tissue was reduced by 22% in STZ-diabetic rats compared with controls. Vanadium treatment did not significantly alter GLUT4 expression in controls, but completely restored normal expression levels in STZ-diabetic rats. In overnight fasted control animals, GLUT4 translocation to the plasma membrane (PM) was maximally elevated (by 50%) in adipose tissue within 5 to 10 minutes after an intravenous (IV) glucose challenge. No glucose-induced translocation of GLUT4 was detected in diabetic rats, and peak PM GLUT4 content was 40% lower than in controls. Vanadium treatment did not increase peak PM GLUT4 content in either control or diabetic animals in response to a glucose load. Finally, the suppression of whole-body acute glucose tolerance in diabetic animals was only partially normalized by vanadium treatment. We conclude: (1) that concentrations of vanadium effective for maintaining normoglycemia in vivo (typically below 30 micromol/L) promote normal GLUT4 expression, but do not influence the subcellular localization of GLUT4 in white adipose tissue and (2) that in vivo effects of vanadium may not necessarily reflect the actions observed in vitro at supraphysiologic concentrations. PMID- 11398145 TI - Interrelationship between estimates of adiposity and body fat distribution with metabolic and hemostatic parameters in obese children. AB - Adiposity in childhood is often associated with metabolic abnormalities and accompanied by a dysregulation of the coagulation and fibrinolytic systems. We studied the interrelationship of metabolic and hemostatic parameters and explored their relationship with measures of adiposity and fat distribution in obese children. In 34 obese boys (mean age, 11.7 years) and 57 obese girls (12.1 years), blood samples were determined for insulin, glucose, triglycerides, fibrinogen, plasminogen activator inhibitor-1 (PAI-1), and tissue-type plasminogen activator-antigen (tPA-Ag). Body composition was assessed by means of impedance. Waist (Wc) and hip circumference were measured. The thickness of subcutaneous adipose tissue-layers (SAT-layers) was measured at 15 different body sites (from 1-neck to 15-calf) by means of the optical device, Lipometer. Overall subcutaneous fatness (SAT) was calculated and SAT-distribution was estimated by means of factor analysis. Significant correlations were found between different measures of adiposity and Wc with metabolic parameters. Fibrinogen was mainly associated with upper body subcutaneous fatness (factor 1) in boys. In girls, hemostatic parameters were associated with nearly all measures of adiposity and also with factor 1 and SAT. Regression analysis showed that factor 1 together with PAI-1 (both P <.0001) contribute to fibrinogen (adjusted [adj], R(2) =.30). PAI-1 together with trigylcerides (both P <.0001) and age (P <.04) were main determinants for tPA-Ag (adj, R(2) =.41). tPA-Ag (P <.0001) together with glucose (P <.001, negative slope), fibrinogen (P <.001, negative slope), and percentage fat mass (%FM) (P <.01) contributed to PAI-1 (adj, R(2) =.54). These results favor the concept of an interrelationship between metabolic and hemostatic parameters resulting from increased adiposity, perhaps influenced by pubertal development of children. Although upper body subcutaneous fatness was found to be a main correlate of metabolic and hemostatic parameters, it remains to be investigated whether this type of subcutaneous fat distribution is involved in the expression of metabolic and hemostatic risk factors and participates in the dysregulation of the hemostatic system in the state of childhood obesity. PMID- 11398146 TI - Gliclazide decreases vascular smooth muscle cell dysfunction induced by cell mediated oxidized low-density lipoprotein. AB - Accumulating evidence indicates that oxidative modification of low-density lipoprotein (LDL) plays an important role in vascular dysfunction associated with diabetes mellitus. The aim of the present study was to investigate the effect of gliclazide, a second-generation sulfonylurea with free-radical-scavenging activity, on human aortic smooth muscle cell (HASMC)-mediated LDL oxidation and HASMC dysfunction induced by oxidatively modified LDL. Incubation of HASMCs with native human LDL (100 microg/mL) in the presence of increasing concentrations of gliclazide (1 to 10 microg/mL) resulted in a dose-dependent decrease in HASMC mediated LDL oxidation. Exposure of HASMCs to gliclazide (1 to 10 microg/mL) and native LDL (100 microg/mL) also led to a dose-dependent decrease in oxidized LDL induced human monocyte adhesion to HASMCs. In addition, incubation of HASMCs with gliclazide dramatically reduced the ability of oxidized LDL to stimulate the proliferation of these cells. Finally, treatment of HASMCs with gliclazide resulted in a marked decrease in oxidatively modified LDL-induced monocyte chemoattractant protein (MCP)-1 and human heat shock protein 70 (hsp 70) expression, both at the gene and protein levels. These results show that gliclazide, at concentrations in the therapeutic range (5 to 10 microg/mL), is effective in vitro in reducing vascular smooth muscle cell (VSMC) dysfunction induced by oxidatively modified LDL. These observations suggest that administration of gliclazide to type 2 diabetic patients could form part of the strategy for the prevention and management of diabetic cardiovascular diseases. PMID- 11398147 TI - Apolipoprotein E polymorphism modulates the association between obesity and dyslipidemias during young adulthood: The Bogalusa Heart Study. AB - To elucidate to what extent apolipoprotein (apo) E polymorphism modulates obesity induced dyslipidemias during young adulthood, longitudinal data on 759 individuals (72% white/28% black; initial and follow-up mean age, 25.9 and 32.7 years) were examined. Among both races and the total sample, the apo E2 group (with E2/2 or E2/3 phenotype) had significantly lower and the apo E4 (with E4/4 or E3/4 phenotype) group higher low-density lipoprotein (LDL) cholesterol than the apo E3 (with E3/3 phenotype) group at both examinations. In addition, the apo E2 group displayed higher high-density lipoprotein (HDL) cholesterol in the total sample. No allele-specific effect was noted for the longitudinal changes (Delta). An increase in Delta adiposity, measured as Delta body mass index (BMI), was accompanied by higher increase in Delta LDL cholesterol in the e4 carriers than the e2 carriers among the whites (P <.05) and the total sample (P <.01); an increase in Delta triglycerides and decrease in Delta HDL cholesterol in the e2 carriers than the e4 carriers among all the groups (P <.05 to.001). Among the apo E phenotype groups, the incidence of high (>75th percentile specific for race and sex) LDL cholesterol at follow-up was in the order E4 > E3 > E2 both in the obese (BMI > 30; P for trend =.033) and the nonobese (BMI < 25; P for trend =.035) groups. Although the increase of low (<25th percentile specific for race and sex) HDL cholesterol or high triglycerides showed no apo E phenotype-specific trend, the incidence of high triglycerides without high LDL cholesterol was in the order E2 > E3 > E4 only in the obese group (P for trend =.025). The prevalence trend for dyslipidemias at follow-up among the persistently obese and nonobese groups also gave similar results. Thus, apo E gene locus influences not only the levels of certain lipoprotein variables during young adulthood, but also modulates the association between obesity and dyslipidemias. PMID- 11398148 TI - Involvement of cyclic guanosine 3',5'-monophosphate in nitric oxide-induced glucagon secretion from pancreatic alpha cells. AB - It has been reported that nitric oxide (NO) is a positive modulator of glucagon release. The involvement of cyclic guanosine 3',5'-monophosphate (cGMP) in NO induced glucagon secretion and the possible role of NO in glucagon release induced by l-arginine were investigated in mouse clonal alpha-cell line clone 6 (alpha TC6) cells, which predominantly secrete glucagon. NOC12, an NO donor, elicited an increase in glucagon release from alpha Tc6 cells in perifusion and static incubation. An inhibitor of cGMP-dependent protein kinase inhibited NOC12 induced glucagon release. NOC12 (1 mmol/L) also increased the cellular level of cGMP. In addition, a permeable cGMP agonist increased glucagon release. l arginine (15 mmol/L) increased perifusate concentrations of glucagon and nitrite in alpha Tc6 cells, which were inhibited by N(G)-nitro-L-arginine methyl ester. NO synthase (NOS) activity was shown in alpha Tc6 cells by l-citrulline formation assay. Our present findings suggest that NO plays a stimulating role in glucagon release from the alpha cells, and that a cGMP-dependent pathway is involved in NO action. These findings also provide further evidence that l-arginine might play a stimulating role in regulating glucagon secretion, at least partly, through generation of NO in the islets. PMID- 11398149 TI - Hepatic cholesterol and bile acid synthesis, low-density lipoprotein receptor function, and plasma and fecal sterol levels in mice: effects of apolipoprotein E deficiency and probucol or phytosterol treatment. AB - We compared hepatic cholesterol metabolism in apolipoprotein (apo) E-knockout (KO) mice with their wild-type counterparts. We also investigated the effects of treatment with phytosterols or probucol on the activity of hepatic 3-hydroxy-3 methyl-glutaryl coenzyme A (HMG-CoA) reductase (cholesterol synthesis), cholesterol 7 alpha-hydroxylase and sterol 27-hydroxylase (bile acid synthesis), and low-density lipoprotein (LDL) receptor function in this animal model of atherogenesis. These findings were then related to treatment-induced changes in plasma, hepatic, and fecal sterol concentrations. Mouse liver membranes have binding sites similar to LDL receptors; the receptor-mediated binding represents 80% of total binding and is LDL concentration-dependent. These binding sites have higher affinity for apo E-containing particles than apo B only-containing particles. Deletion of apo E gene was associated with several-fold increases in plasma cholesterol levels, 1.5-fold increase in hepatic cholesterol concentrations, 50% decrease in HMG-CoA reductase activity, 30% increase in cholesterol 7 alpha-hydroxylase and 25% decrease in LDL receptor function. Treatment of apo E-KO mice with either probucol or phytosterols significantly reduced plasma cholesterol levels. Phytosterols significantly increased the activity of hepatic HMG-CoA reductase, and probucol significantly increased cholesterol 7 alpha-hydroxylase activity. Neither treatment significantly altered hepatic LDL receptor function. Phytosterols, but not probucol, significantly increased fecal sterol excretion and decreased hepatic cholesterol concentrations. Plasma cholesterol lowering effects of phytosterols and probucol are due to different mechanisms: stimulation of cholesterol catabolism via increased bile acid synthesis by probucol and decreased cholesterol absorption by phytosterols. In the absence of apo E, hepatic LDL receptors could not be upregulated and did not contribute to the cholesterol lowering effects of either agent. PMID- 11398150 TI - Beta-cell function and visceral fat in lactating women with a history of gestational diabetes. AB - Lactation has been recommended as beneficial for the maternal metabolic abnormalities associated with glucose intolerance and diabetes risk, although associations between breastfeeding (BF), glucose tolerance, and adipose tissue distribution are unknown. Therefore, a population of women with recent gestational diabetes (GDM) was evaluated with comparison of results for lactating versus nonlactating women. A total of 26 women participated (14 BF and 12 nonbreastfeeding [nonBF]) with a singleton vaginal delivery after 36 weeks gestation. At 3 months postpartum, each woman completed a 75-g oral glucose tolerance test (OGTT), a frequently sampled intravenous glucose tolerance test (FSIGT), and computed tomography (CT) scanning for adipose distribution and mass. Insulin sensitivity was not significantly different between BF and nonBF groups (4.97 +/- 0.78 v 3.44 +/- 1.0 x 10(-4) min(-1)/(microU/mL) nor was glucose effectiveness (1.92 +/- 0.22 v 1.56 +/- 0.19 x 10(-2) min(-1)). However, the disposition index (DI) (insulin sensitivity [S(I)] x acute insulin response to glucose [AIRg]) was higher in the BF group (129.9 +/- 26.0 v 53.4 +/- 18.0 x 10( 4) min(-1); P =.03). Visceral fat (103 +/- 14 v 97 +/- 15 cm(2)) and subcutaneous fat (362 +/- 36 v 460 +/- 68 cm(2)) were similar between the groups. We conclude that 3 months of BF in a population with previous GDM was associated with improved pancreatic beta-cell function, but not with any difference in measures of adiposity. PMID- 11398151 TI - Amino acid, glucose, and lipid kinetics after palliative resection in a patient with glucagonoma syndrome. AB - Glucagon excess causes catabolic changes, including enhanced glucose production, lipolysis, and amino acid oxidation. In this study, we evaluate the metabolic effects of debulking surgery on a patient with glucagon-producing tumor. Stable isotope tracer methods were used to measure glucose, glycerol, and alpha ketoisocaproic acid (alpha KICA) rates of appearance (Ra) into plasma. Measurements were obtained 25 days after surgery in the basal state and during hormonal suppression of glucagon production by infusing somatostatin with insulin replacement. Basal plasma glucagon concentration (14,100 pg/mL) remained high after debulking surgery. Somatostatin infusion decreased plasma glucagon concentration to 6,735 pg/mL and basal substrate kinetics (alpha-KICA Ra from 1.97 to 1.48 micromol/kg/min; glucose Ra from 16.89 to 11.56 micromol/kg/min; and glycerol Ra from 3.33 to 2.74 micromol/kg/min). We conclude that debulking surgery fails to adequately suppress glucagon production and the alterations in substrate metabolism associated with excess glucagon. In these patients, somatostatin therapy can be an effective method to suppress secretion of glucagon and help attenuate its catabolic effects. PMID- 11398152 TI - Lipoprotein profile and cholesteryl ester transfer protein in neonates. AB - Undernourishment in utero appears to be associated with persisting changes in the metabolic, endocrine, and immune functions. In this study, we determined the influence of birth weight on the lipoprotein profile and cholesteryl ester transfer protein (CETP), which promotes a proatherogenic lipoprotein profile in plasma by determining the chemical, physical, and biologic properties of the respective lipoprotein particles. Triglyceride (TG) concentrations were highest and high-density lipoprotein (HDL)(2)-cholesterol levels were lowest in small for gestational age (SGA) neonates. CETP-mass was determined by enzyme-linked immunosorbent assay (ELISA) and CETP-activity by using exogenous lipoproteins. Cholesteryl ester transfer was determined as transfer of radiolabeled cholesteryl esters (CE) from HDL to apolipoprotein B-containing lipoproteins. CETP mass was lowest and cholesteryl ester transfer was highest in SGA neonates. CETP-activity did not differ among the neonates. Our results suggest that increased and decreased nourishment in utero affects the lipoprotein profile and CETP in neonates. High TG and low HDL(2) levels in SGA neonates might result from increased cholesteryl ester transfer and, may in part, explain the increased risk of coronary heart disease (CHD) of small for gestational age neonates in later life. PMID- 11398153 TI - Ciprofibrate versus gemfibrozil in the treatment of mixed hyperlipidemias: an open-label, multicenter study. AB - Mixed hyperlipidemia is a common risk factor for cardiovascular disease. The aim of this trial was to evaluate the efficacy and safety of ciprofibrate versus gemfibrozil for the treatment of patients with mixed hyperlipidemia carefully selected for similar lipid profiles. A total of 68 patients who had mixed hyperlipidemia after following an isocaloric American Heart Association (AHA) phase I diet for 4 weeks were included. The plasma lipid levels at the inclusion were low-density lipoprotein-cholesterol (LDL-C) > or = 130 mg/dL, cholesterol > or = 240 mg/dL, and triglycerides > or = 200 mg/dL. Patients were randomly assigned to receive ciprofibrate 100 mg/d or gemfibrozil 1,200 mg/d. At the end of the 8-week treatment period, efficacy and safety parameters were compared with baseline values. The primary efficacy parameters of the study were percentage changes in triglycerides and LDL-C from baseline. After 8 weeks, plasma triglyceride concentrations were decreased by 43.5% and 54% compared with baseline during ciprofibrate and gemfibrozil therapy, respectively (P <.001). High-density lipoprotein-cholesterol (HDL-C) concentrations were increased 20.8% and 19.3% during ciprofibrate and gemfibrozil, respectively (P <.001). Apoprotein B, cholesterol, and very-low-density lipoprotein-cholesterol (VLDL-C) concentrations were also improved by the study drugs (18.6%, 13.2%, and 30.9%, respectively, during ciprofibrate and 44%, 13.8%, and 14.4%, respectively, during gemfibrozil). Meanwhile, the effect of the drug was minimal on LDL-C. A significant decrease in non-HDL-C resulted from both treatments (19% and 19.5%, respectively, P <.05). The only statistically significant difference observed between treatments was the effects on fibrinogen concentration, a coronary risk factor. Ciprofibrate significantly decreased its concentration by 18.8%, fibrinogen was slightly increased during gemfibrozil treatment. No patient had a significant modification on any of the safety tests. In summary, ciprofibrate and gemfibrozil are well-tolerated and efficacious treatments for mixed hyperlipidemia. Significant reductions in triglycerides, non-HDL-C, and apolipoprotein B were achieved with both drugs. A significant fibrinogen reduction was obtained with ciprofibrate. PMID- 11398154 TI - Increase of lipogenic enzyme mRNA levels in rat white adipose tissue after multiple cycles of starvation-refeeding. AB - Recently, we have found that despite the significant reduction of body weight after multiple starvation-refeeding cycles, white adipose tissue (WAT) exhibits surprisingly high rates of lipogenesis and lipogenic enzyme activities. The purpose of this study was to determine the response of WAT lipogenic enzyme mRNAs of rats subjected to multiple cycles of 3 days fasting and 3 days of refeeding. Despite the body weight reduction, significant increase of lipogenic enzymes (ie, fatty acid synthase [FAS], acetyl-coenzyme A [CoA] carboxylase [ACC], adenosine triphosphate (ATP)-citrate lyase [ACL], NADP-linked malic enzyme [ME], and glucose 6-phosphate dehydrogenase [G6PDH]) mRNAs in WAT was found after multiple cycles of starvation-refeeding of rats on standard laboratory diet. These findings, together with the results published recently, indicate that multiple cycles of starvation-refeeding cause the increased lipogenesis in WAT by upregulation of the lipogenic enzymes gene expression. PMID- 11398156 TI - Skin cancer and ozone depletion: the case for global action. PMID- 11398157 TI - Pelvic soft tissue sarcomas present a challenge to therapists. PMID- 11398158 TI - Technical issues and pathologic implications of sentinel lymph node biopsy in early-stage breast cancer patients. AB - BACKGROUND AND OBJECTIVES: Recent studies have demonstrated that the sentinel lymph node (sN) can be considered a reliable predictor of axillary lymph node status in breast cancer patients. However, some important issues, such as optimization of the technique for the intraoperative identification of the sN, and the clinical implications of sN metastasis as regards the surgical management of the axilla still require further elucidation. The objectives of this study was to assess (1) the feasibility of sN identification with a combined approach (vital blue dye lymphatic mapping and radioguided surgery, RGS) and the specific contribution of either techniques to the detection of the sN, and (2) the correlation between the size of sN metastasis (micrometastasis < or = 2 mm; macrometastasis > 2), primary tumour size, and the status of nonsentinel nodes (nsN) in the axilla. METHODS: Between October of 1997 and December of 1999, 212 patients with breast cancer (average age: 61 years; range, 40-79 years) underwent sN biopsy before performing standard axillary dissection. In a subset of 153 patients, both vital blue dye (Patent Blue-V) lymphatic mapping and RGS were used to identify the sN, and the relative contribution of each of the two techniques was assessed. RESULTS: Overall, the sN was identified in 206 of 212 patients (97.1%); at histologic examination of all dissected nodes, 77 of 206 patients had positive nodes (37.3%). The false-negative rate was 6.5% (5/77), the negative predictive value was 96.3% (129/134), and accuracy was 97.6% (201/206). Among 72 patients with positive sN, micrometastases were detected in 21 cases and macrometastases in 51. When micrometastases only were observed, the sN was the exclusive site of nodal metastasis in 17 of 21 cases (80.9%); in the remaining 4 cases (19.1%), nsN metastases were detected in 3 of 14 pT1c patients (21.5%), and 1 of 5 pT2 patients (20%). Macrometastases were detected in patients with tumors classified as pT1b or larger: the sN was the exclusive site of metastasis in 3 of 4 pT1b patients (75%), in 14 of 29 pT1c patients (48.2%), and in 3 of 18 pT2 patients (16.6%). The specific contribution of the two different techniques used in the identification of the sN was evaluated; the detection rate was 73.8% (113 of 153) with Patent Blue-V alone, 94.1% (144 of 153) with RGS alone, and 98.7% (151 of 153) with Patent Blue-V combined with RGS (P < 0.001). Noteworthy, whenever the sN was identified, the prediction of axillary lymph node status was remarkably similar (93-95% sensitivity; 100% specificity; 95-97% negative predictive value, and 97-98% accuracy) with each of the three procedures (Patent Blue-V alone, RGS alone, or combined Patent Blue-V and RGS). CONCLUSIONS: Sentinel lymphadenectomy can better be accomplished when both procedures (lymphatic mapping with vital blue dye and RGS) are used, due to the significantly higher sN detection rate, although the prediction of axillary lymph node status remains remarkably similar with each one of the methods assessed. That patients with small tumours (<1 cm) and sN micrometastasis are very unlikely to harbour metastasis in nsN should be considered when planning randomised clinical trials aimed at defining the effectiveness of sN guided-axillary dissection. PMID- 11398160 TI - Comparison of two methods of reconstruction for primary malignant tumors at the knee: a sequential cohort study. AB - BACKGROUND AND OBJECTIVES: The purpose of this study was to compare the complications and functional outcome associated with the use of an irradiated allograft-implant composite or a bone-ingrowth modular tumor prosthesis for replacement of the knee joint after resection of a bone sarcoma from the distal femur or proximal tibia. METHODS: Eleven patients initially received an allograft reconstruction, followed by 64 treated with a tumor prosthesis. The primary analysis concerned reconstructive failure, defined by the requirement for removal of the original construct. Functional outcome was assessed by using the 1987 Musculoskeletal Tumor Society rating system. RESULTS: Reconstructive failure occurred in 6 of 11 (55%) allograft constructs compared with 10 of 64 (16%) tumor prostheses (P = 0.009). Failures were due to infection (2 of 11 allografts versus 4 of 64 prostheses; P = 0.2) or mechanical complications (4 of 11 allograft fractures versus 5 of 64 broken prosthetic stems and 1 aseptically loose prosthesis; P = 0.03). The limb salvage rate was 95% (61 of 64) for patients with a tumor prosthesis compared with 64% (7 of 11) for those with an allograft (P = 0.007). Patients with a tumor prosthesis had a better functional outcome with a mean score of 75% compared with 57% for those with an allograft reconstruction (P = 0.006). CONCLUSIONS: This comparative study suggests that limb salvage surgery at the knee has a better and more predictable outcome with a tumor prosthesis than with an allograft-implant reconstruction. PMID- 11398162 TI - Adult perineal sarcomas. AB - BACKGROUND AND OBJECTIVES: Primary perineal sarcoma in adults is a rare disease that has only been documented to occur in isolated case reports. METHODS: To better characterize and define the natural history of perineal sarcoma in adults (> or = 18 years), we reviewed our experience with treatment of perineal sarcoma between 1982 and 1999 (nine cases). RESULTS: Epithelioid sarcoma (n = 4) was the most common histologic subtype. Seven cases (78%) were histologically high grade, and lesions were most commonly < 5 cm. All patients were treated with wide local excision. External beam radiation was the most commonly used form of adjuvant therapy (n = 6). Recurrences were noted in five patients, and the recurrences were most commonly local (60%). Median time to first recurrence was 21 months. Six of nine patients are alive with a median follow-up of 54 months. Three died of recurrent/metastatic disease at 16, 51, and 54 months after initial surgery. CONCLUSIONS: Aggressive therapy and follow-up beginning with wide excision can be associated with long-term survival in adults with primary perineal sarcoma. PMID- 11398163 TI - Palliative forequarter amputation for metastatic carcinoma to the shoulder girdle region: indications, preoperative evaluation, surgical technique, and results. AB - BACKGROUND AND OBJECTIVES: Uncontrolled metastatic carcinoma of the shoulder girdle is a difficult oncologic problem. This study reviews our experience with palliative forequarter amputation with emphasis on patient selection criteria, preoperative radiologic assessment, surgical technique, epineural postoperative analgesia, and clinical outcome. METHODS: Eight patients who underwent palliative forequarter amputation for metastatic carcinoma between 1980 and 1999 were analyzed retrospectively. Diagnoses included breast carcinoma (n = 3), squamous cell carcinoma (n = 2), hypernephroma (n = 2), and carcinoma of unknown origin (n = 1). All patients presented with severe, intractable pain and a useless extremity. Venography demonstrated obliteration of the axillary vein in each of the patients in whom this procedure was performed. Exploration of the brachial plexus confirmed tumor encasement and unresectability in all patients. Epineural catheters for bupivacaine infusion were placed for postoperative pain control. RESULTS: All patients experienced dramatic pain relief and improved mobility and overall function. Life-threatening hemorrhage and sepsis were alleviated. There were no instances of phantom limb pain or adverse psychological reactions, and no complications related to epineural analgesia. CONCLUSIONS: Palliative forequarter amputation is relatively safe and reliable and provides effective pain relief for selected patients with unresectable metastatic carcinoma to the axilla and bony shoulder girdle in whom radiotherapy and/or chemotherapy has not been effective. The triad of pain, motor loss, and an obliterated axillary vein is indicative of brachial plexus infiltration and unresectability. PMID- 11398165 TI - Diminished benefit from resection of cancer of the head of the pancreas in patients of advanced age. AB - BACKGROUND AND OBJECTIVES: The incidence of pancreatic cancer is increasing, and an increasing proportion of these patients is older than 65 years. The benefits of resection in the geriatric population, in whom major comorbidity is more likely, are poorly defined. The authors sought to determine the relative benefits of resection of cancer of the head of the pancreas in different age groups, with particular emphasis on the geriatric population. METHODS: Between 1983 and 1995, 273 patients presented to the University of Miami for evaluation of noncystic epithelial cancer of the head of the pancreas. Resection was performed in 104 patients, and these patients are the subject of this retrospective review. Mean length of follow-up for surviving patients was 37 +/- 24 months. Outcomes were compared in patients < 65 years old (group 1, n = 38), 65-74 years old (group 2, n = 47), and > 74 years old (group 3, n = 19). RESULTS: Total pancreatectomy was performed in 12 patients and pancreaticoduodenectomy was performed in 92 patients. The overall complication rate was similar in all groups, but major morbidity was highest in group 3 (P = 0.05). Median survival for patients in group 2 was 25.1 months. Survival was significantly shorter in patients from groups 1 and 3 (median survivals 12.4 months and 11.4 months, respectively; P = 0.02). Following control for Hispanic ethnicity, which was also a significant prognostic factor on univariate analysis, only the oldest age group had a significantly shorter survival than the other two groups. Age > 74 years and Hispanic ethnicity remained significant after multivariate analysis. CONCLUSIONS: Long-term survival after resection is truncated in older patients. This finding and the observation that the major complication rate is higher in the older subgroup emphasize the need to evaluate critically whether older patients should be submitted to radical resection. PMID- 11398166 TI - Cell proliferative activity and expression of cell-cell adhesion factors (E cadherin, alpha-, beta-, and gamma-catenin, and p120) in sarcomatoid renal cell carcinoma. AB - BACKGROUND AND OBJECTIVES: Sarcomatoid RCC (renal cell carcinoma) is an uncommon, but not rare, neoplasm which has been shown to have a much worse prognosis than common RCC. The current study was designed to investigate the association of proliferative activity and cell-cell adhesion molecules with sarcomatoid RCC. METHODS: Proliferative activity (Ki-67 labeling index) and expression of cell cell adhesion associated molecules (E-cadherin, alpha-, beta-, and gamma-catenin, and p120) were examined using immunohistochemical techniques in 11 cases of sarcomatoid RCC. RESULTS: In six patients with sarcomatous component more than 50%, five were died within 24 month after diagnosis. The expression of these molecules within the carcinomatous and sarcomatous components of sarcomatoid RCC was compared. The mean Ki-67 labeling index in the sarcomatous components (12.6%) is statistically higher than in the carcinomatous components (3.7%) (P < 0.05). The expressions of E-cadherin, and alpha-, beta-, and gamma-catenin were statistically decreased in the sarcomatous components compared to the carcinomatous components. However, no differences were observed regarding p120 immunostaining. CONCLUSIONS: The findings of the current study suggest that the range of the sarcomatous component may be a prognostic factor in RCC, and the malignant behavior of sarcomatoid RCC is due to high cell proliferative activity and decreased expressions of E-cadherin and alpha-, beta-, and gamma-catenin in the sarcomatous component. PMID- 11398167 TI - Central pancreatectomy for tumors of the neck and body of the pancreas. PMID- 11398168 TI - Intrathyroid parathyroid carcinoma. PMID- 11398169 TI - Biologic basis and evolving role of aromatase inhibitors in the management of invasive carcinoma of the breast. AB - BACKGROUND AND OBJECTIVES: The most powerful predictor of the response of breast cancers to hormonal therapy is the presence of estrogen receptors in the tumor cells. Estrogen receptors are expressed in approximately 35-55% of all breast tumors but up to 80-90% of tumors from women older than 55 years. METHODS: At this time, tamoxifen remains the first-line hormonal therapy for breast cancer of all stages. However, the aromatase inhibitors are evolving into an important treatment option. Aromatase inhibitors prevent the conversion of precursors (androgens) to estrogens. RESULTS: On the basis of several randomized clinical trials, aromatase inhibitors have become established as the second-line therapy for postmenopausal women with advanced breast cancer progressing during tamoxifen therapy. Furthermore, very recent trials support the use of these agents as first line therapy in place of tamoxifen. CONCLUSIONS: The roles of the selective aromatase inhibitors in the prevention of breast cancer and in the neoadjuvant and adjuvant treatment of early-stage breast cancer are the focus of several planned and ongoing large-scale clinical trials. These trials will answer some of the many questions that remain regarding optimal hormonal therapy for hormone dependent breast cancer. PMID- 11398170 TI - Comparison of prazosin, terazosin and tamsulosin: functional and binding studies in isolated prostatic and vascular human tissues. AB - BACKGROUND: Terazosin and tamsulosin are drugs currently used in the treatment of benign prostatic hypertrophy (BPH). The potency of these two alpha(1) receptor antagonists and that of prazosin to inhibit contractions induced by noradrenaline and the binding of [(3)H]-prazosin in human prostate and four different human arterial and venous vessels (saphenous and umbilical veins, renal and mesenteric arteries) was studied. METHODS: By bioassay and binding studies, we examined the receptor affinities of different alpha(1) receptor antagonists in different human tissues. RESULTS: pKb of terazosin, tamsulosin, and prazosin obtained in the prostatic tissues (8.15, 9.64, and 8.59, respectively) were not different from those obtained in the umbilical veins (8.07, 9.56, and 8.30, respectively), in the mesenteric artery (8.27, 10.29, and 9.01, respectively), renal artery (8.35, 10.13, and 8.76, respectively) and saphenous vein (7.8, 10.3, and 9.32, respectively). IC(50) (nM) of prazosin, terazosin, and tamsulosin obtained from binding studies in membrane preparations from prostate tissue were similar to those from umbilical veins, saphenous vein, and renal artery. CONCLUSIONS: All of the evaluated drugs showed similar selectivity for prostatic vs. vascular tissues. Thus, different clinical profiles of the present drugs should not result from their differential affinity for prostatic versus vascular alpha(1) adrenoceptors. PMID- 11398171 TI - Phage display selection of peptides that affect prostate carcinoma cells attachment and invasion. AB - BACKGROUND: Prostate cancer-specific proteins must be identified to serve as diagnostic and prognostic markers. Cell surface proteins are especially important, because they have potential utility as diagnostic markers and therapeutic targets. Identification of ligands for these proteins will allow use of these ligands as diagnostic and therapeutic tools and permit the investigation of receptor function. We performed a search for peptide ligands to prostate cancer cell-specific receptors. METHODS: Peptide phage display library was used to isolate specific ligands to LNCaP prostate carcinoma cells receptors. Selected phage and cognate peptides were investigated for their cancer-related functions, such as the ability to interfere with cell adhesion, spreading, motility, and invasion. RESULTS: Phage designated pg35, blocked spreading of LNCaP cells and their derivatives C4-2 and C4-2b. Cognate peptide did not inhibit spreading, but incubation of C4-2 and C4-2b cells with cognate peptide increased their affinity for endothelial cells and invasiveness. In addition, the peptide activates matrix metalloproteinase (MMP)-2 and 9 in C4-2 and C4-2b cells. CONCLUSIONS: These results indicate that identified ligands may play a role in tumorigenicity and metastatic transformation of prostate cancer. To our knowledge, this is the first identification of a functional cancer-specific peptide ligand using the phage display approach. PMID- 11398172 TI - Tumor characteristics in screening for prostate cancer with and without rectal examination as an initial screening test at low PSA (0.0-3.9 ng/ml). AB - BACKGROUND: The value of rectal examination as initial screening test for prostate cancer at low PSA values (0.0-3.9 ng/ml) was determined by evaluating the number and tumor characteristics of the cancers detected. METHODS: Two study populations were subjected to screening with (n = 10,226) and without (n = 10,753) rectal examination as initial screening test. The number of cancers detected at low PSA values for both screening regimens, the corresponding biopsy and radical prostatectomy tumor characteristics were assessed. Possibly harmless cancers were defined as small (< 0.5 ml) organ-confined tumors without Gleason growth-patterns 4/5. RESULTS: At low PSA, 26.6% (117/440) of screen-detected cancers were detected after the evaluation of a suspicious rectal examination. The number of cancers and tumor aggressiveness features were highly associated with serum-PSA level. The proportion of possibly harmless disease steadily declined from 100% (PSA 0.0-0.9 ng/ml) to 15.4% (PSA 3.0-3.9 ng/ml). Rectal examinations were performed unnecessarily in 94.7-100% of cases, when detection of clinically significant disease was aimed at. Using PSA (and a cut-off of 3.0 ng/ml) as the only screening tool, 24.3% (121/498) of screen-detected cancers were in the PSA range 3.0-3.9 ng/ml, and 60.0% were assessed as clinically significant. CONCLUSIONS: Rectal examination as initial screening test for prostate cancer at low PSA values may be replaced by screening using serum-PSA only. At PSA levels below 3.0 ng/ml, 289 rectal examinations are required to find one case of clinically significant disease, and 96 rectal examinations are needed to diagnose prostate cancer of any size, grade, or stage. PMID- 11398173 TI - The association of fatty acids with prostate cancer risk. AB - BACKGROUND: Animal studies indicate that omega-6 fatty acids promote and omega-3 fatty acids inhibit tumor development. This pilot study was designed to evaluate whether these fatty acids are associated with human prostate cancer. METHODS: Levels of erythrocyte membrane omega-3 and omega-6 fatty acids were determined for 67 incident prostate cancer cases and 156 population-based controls. RESULTS: Prostate cancer risk was increased in the highest compared to the lowest quartile of alpha-linolenic acid (OR = 2.6, 95% CI = 1.1-5.8, trend P = 0.01). Positive associations were also observed with higher levels of linoleic acid (OR = 2.1, 95% CI = 0.9-4.8) and total omega-6 fatty acids (OR = 2.3, 95% CI = 1.0-5.4). CONCLUSIONS: Results are consistent with other studies showing that linoleic and total omega-6 fatty acids increase risk of prostate cancer. Contrary to animal studies, alpha-linolenic acid was also positively associated with risk. Further research will be required to clarify the role of these fatty acids in human prostate cancer. PMID- 11398175 TI - Gonadotropin-releasing hormone receptor expression in the human prostate. AB - BACKGROUND: Inhibitory effects of gonadotropin-releasing hormone (GnRH) analogs on prostate cancer cell proliferation, both in vivo and in vitro, indicate the presence of specific binding sites for GnRH on prostate cancer cells. To investigate this issue further, we examined the expression of GnRH receptor (GnRH R) mRNA and protein in human prostate biopsies as well as in other extrapituitary tissues. METHODS: The relative quantity of GnRH-R mRNA was determined by reverse transcriptase-polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC) in human prostate biopsies. Extrapituitary GnRH-R levels were determined by a semiquantitative PCR reaction. RESULTS: Using PCR, a relatively high expression level of GnRH-R mRNA was found in prostate tumor tissue followed by normal prostate, thymus, and kidney expression levels. The levels showed by heart, brain, placenta, lung, liver, skeletal muscle, pancreas, colon, ovary, small intestine, spleen, and testis were low but detectable, whereas peripheral blood leukocyte showed no demonstrable product. GnRH-R immunoreactivity was localized in both luminal and basal epithelial cells in benign and malignant prostate tissue, and GnRH-R were also observed in intraprostatic lymphocytes. The relative GnRH-R mRNA levels in prostate biopsies from 16 patients showed a wide range of individual differences, but these differences were not related to histological grade. Castration therapy did not significantly influence GnRH-R mRNA expression in normal and malignant prostate tissue. CONCLUSIONS: These results suggest that epithelial cells and infiltrating lymphocytes are targets for GnRH action in the human prostate. Comparative data show relatively high GnRH-R expression in human prostate tissue compared to other human tissues. PMID- 11398174 TI - Reduced expression of hMSH2 and hMLH1 and risk of prostate cancer: a case-control study. AB - BACKGROUND: Although prostate cancer is the most common incident cancer in men, not much is known about its etiology. We tested the hypothesis that expression levels of hMSH2 and hMLH1 in unaffected (normal) tissue play a role in the etiology of prostate cancer. METHODS: Total RNA was extracted from peripheral blood lymphocytes of subjects ascertained by a case-control study (70 patients and 97 age- and ethnicity-matched controls). A multiplex reverse transcription polymerase chain reaction assay was used to simultaneously evaluate the relative expression of hMSH2 and hMLH1, using beta-actin as the internal control. RESULTS: The relative gene expression levels of hMSH2 and hMLH1 were significantly lower in cases than in controls (P < 0.05 for both genes). When compared with the highest tertile of the controls, low expression levels (the middle and lowest tertiles) of hMLH1 were associated with significantly increased risk of prostate cancer in a dose-response relationship (ORs = 2.68, and 4.31; 95% confidence interval = 1.00-7.23 and 1.64-11.30, respectively) after adjustment for age, ethnicity, smoking status, and family history of prostate cancer. CONCLUSIONS: These results suggest that reduced expression of hMLH1 in peripheral lymphocytes may be a risk factor for prostate cancer. However, it cannot be ruled out that the reduced expression we observed may be caused by the disease status. Our findings and the factors that may affect the expression of hMLH1 need further confirmation in larger prospective studies. PMID- 11398176 TI - Vasoactive intestinal peptide (VIP) stimulates rat prostatic epithelial cell proliferation. AB - BACKGROUND: Androgens play a major role in supporting normal growth and functional maintenance in the prostate. However, this gland contains an array of neuroendocrine peptides that can play a regulatory role in its physiopathology. Among these peptides, one of the best studied is vasoactive intestinal peptide (VIP), which is abundant in autonomic nerves surrounding both human and rat prostatic acini. This neuropeptide may act through interaction with two types of high-affinity receptors, named VPAC(1) and VPAC(2) receptors. Another regulatory peptide, the pituitary adenylate cyclase-activating peptide (PACAP), interacts with these receptors with the same affinity as VIP, but binds with higher affinity to PAC(1) receptors. Human prostate tumors and rat prostate show a major presence of VPAC(1) receptors, whereas various findings suggest a role for VIP in prostatic development. Here we studied the effects of VIP on the proliferation of rat prostatic epithelial cells in culture. METHODS: We studied the [(3)H] thymidine uptake by rat prostatic epithelial cells in culture, characterized previously by using biomarkers such as cytokeratin and vimentin. In these cells we tested the effect of VIP and PACAP-27 on two different signaling pathways, the cyclic AMP (cAMP) and the inositol phosphate (IPs). RESULTS: The rat prostatic cells in culture were cytokeratin (5,6,8) and vimentin positive, indicating that the culture was predominantly epithelial. The proliferation curves showed that the cells followed different states of growth: a quiescent, an exponential proliferative, and a steady state. Cyclic AMP production, but not inositol phosphate production, was increased in the presence of VIP and PACAP-27, which suggests the expression of VPAC(1) and/or VPAC(2) receptors primarily. VIP significantly increased prostatic cell proliferation in a bimodal manner, as shown for dibutyryl cyclic AMP (dbcAMP), which suggests that the effect of VIP upon prostatic proliferation is cAMP-dependent. CONCLUSIONS: Here, we demonstrate that VIP increased [(3)H]thymidine uptake by rat prostatic epithelial cells in culture, conceivably by the activation of the adenylate cyclase. PMID- 11398177 TI - Therapeutic potential of curcumin in human prostate cancer. III. Curcumin inhibits proliferation, induces apoptosis, and inhibits angiogenesis of LNCaP prostate cancer cells in vivo. AB - BACKGROUND: Earlier work from our laboratory highlighted the therapeutic potential of curcumin (turmeric), used as a dietary ingredient and as a natural anti-inflammatory agent in India and other Southeast Asian countries. This agent was shown to decrease the proliferative potential and induce the apoptosis potential of both androgen-dependent and androgen-independent prostate cancer cells in vitro, largely by modulating the apoptosis suppressor proteins and by interfering with the growth factor receptor signaling pathways as exemplified by the EGF-receptor. To extend these observations made in vitro and to study the efficacy of this potential anti-cancer agent in vivo, the growth of LNCaP cells as heterotopically implanted tumors in nude mice was followed. METHODS: The androgen-dependent LNCaP prostate cancer cells were grown, mixed with Matrigel and injected subcutaneously into nude mice. Experimental group received a synthetic diet containing 2% curcumin for up to 6 weeks. At the end point, sections taken from the excised tumors were evaluated for pathology, cell proliferation, apoptosis, and vascularity. RESULTS: Curcumin causes a marked decrease in the extent of cell proliferation as measured by the BrdU incorporation assay and a significant increase in the extent of apoptosis as measured by an in situ cell death assay. Moreover, a significant decrease in the microvessel density as measured by the CD31 antigen staining was also seen. CONCLUSIONS: Curcumin could be a potentially therapeutic anti-cancer agent, as it significantly inhibits prostate cancer growth, as exemplified by LNCaP in vivo, and has the potential to prevent the progression of this cancer to its hormone refractory state. PMID- 11398178 TI - Lipopolysaccharide-induced switch between retinoid receptor (RXR) alpha and glucocorticoid attenuated response gene (GARG)-16 messenger RNAs in cultured rat microglia. AB - Glucocorticoid-attenuated response genes (GARG) belong to a recently described family of genes responsive to the action of dexamethasone. Full-length cDNA of one member of this family, GARG16, has been cloned from rat microglia and regulation of its mRNA expression has been studied. Moreover, regulation of retinoid/retinoic acid activated transcription factor (RXR/RAR) mRNAs in mixed astrocyte and in purified microglia cultures has been investigated. RARbeta mRNA was undetectable in microglia by RT-PCR, whereas clearly present in the mixed cultures. RXRalpha, RARgamma, and GARG16 mRNAs were found in both culture systems. RXRalpha mRNA was strongly expressed in control microglia but rapidly declined upon treatment with LPS. Conversely, GARG16 mRNA was almost untraceable in control microglia but rapidly increased by LPS. Time-course studies revealed an oscillating behavior of expression of both mRNAs during the first 6 hr, which receded to control levels (RXRalpha high, GARG16 low) at 72 hr of LPS-treatment. Additionally, p38 MAPK and SEK phosphorylations peaked at 1 hr followed by steady declines, whereas MEK and c-Jun showed double peaks at 1+4 hr and 1+6 hr, respectively, before subsiding to control levels. This behavior was not observed in comparative studies with TNF-alpha, interleukin-10 (IL-10), or interferon gamma inducible protein 10 (IP-10). Finally, inhibitors of p38 MAPK, p42/p44 ERK, and PKCalpha as well as the use of dexamethasone revealed major influences of the p38 MAPK-c-Jun-AP-1 signaling pathway on RXRalpha and GARG16 mRNA expressions. The counter regulatory control of GARG16 and RXRalpha mRNA expression is believed to be an example of a fine-tuned cellular mechanism to react to inflammatory stimuli. PMID- 11398179 TI - Stimulation of myelin gene expression in vitro and of sciatic nerve remyelination by interleukin-6 receptor-interleukin-6 chimera. AB - Induction of myelin gene expression denotes the last stage of differentiation of myelinating glial cells. Following peripheral nerve transection, Schwann cells (SC) lose myelin gene expression and proliferate, resembling premyelinating embryonic SC (eSC). We show that a fusion protein of the soluble interleukin-6 receptor to interleukin-6 (IL6RIL6), a potent activator of the gp130 signaling receptor, is an inducer of MBP and Po gene products in rat E18 embryonic dorsal root ganglia (DRG) 3 day cultures. Cells whose growth is dependent on the IL6RIL6 chimera were isolated from DRG. These cells (designated CH cells) express Krox 20, as do promyelinating and myelinating SC (mSC). IL6RIL6 induces Po and MBP in CH cells and their cocultures with neurons. In addition, IL6RIL6 leads to a disappearance of Pax-3, a marker of eSC and nonmyelinating Schwann cells (nmSC). Glial fibrillary acidic protein, present in nmSC, is not significantly induced by IL6RIL6. The CH cells acquire glial morphology when exposed to IL6RIL6 and cover axons in cocultures. In a sciatic nerve-derived SC line, IL6RIL6 also induces Po and triggers a rapid attachment along axons. In vivo administration of IL6RIL6 intraperitoneally to rats after sciatic nerve transection and resuture increases 4-fold the number of myelinated nerve fibers (MF) measured on day 12, 2.5-5 mm distal to the suture. The stimulation by IL6RIL6 treatment is highest (7.1-fold) at the more distant 5 mm site, and the thickness of myelin sheaths is increased. Compared to known SC growth factors, the gp130 activator IL6RIL6 appears to combine both in vitro mitogenic effects and promotion of myelin gene expression. PMID- 11398180 TI - Alterations in ciliary neurotrophic factor signaling in rapsyn deficient mice. AB - Rapsyn is a key molecule involved in the formation of postsynaptic specializations at the neuromuscular junction, in its absence there are both pre- and post-synaptic deficits including failure to cluster acetylcholine receptors. Recently we have documented increases in both nerve-muscle branching and numbers of motoneurons, suggesting alterations in skeletal muscle derived trophic support for motoneurons. The aim of the present study was to evaluate the contribution of target derived trophic factors to increases in motoneuron branching and number, in rapsyn deficient mice that had their postsynaptic specializations disrupted. We have used reverse transcription-polymerase chain reaction and Western blot to document the expression of known trophic factors and their receptors in muscle, during the period of synapse formation in rapsyn deficient mouse embryos. We found that the mRNA levels for ciliary neurotrophic factor (CNTF) was decreased in the rapsyn deficient muscles compared with litter mate controls although those for NGF, BDNF, NT-3 and TGF-beta2 did not differ. We found that both the mRNA and the protein expression for suppressor of cytokine signaling 3 (SOCS3) decreased although janus kinase 2 (JAK2) did not change in the rapsyn deficient muscles compared with litter mate controls. These results suggest that failure to form postsynaptic specializations in rapsyn deficient mice has altered the CNTF cytokine signaling pathway within skeletal muscle, the target for motoneurons. This alteration may in part, account for the increased muscle nerve branching and motoneuron survival seen in rapsyn deficient mice. PMID- 11398181 TI - Chemokine antagonist infusion promotes axonal sparing after spinal cord contusion injury in rat. AB - Spinal cord injury produced by mechanical contusion causes the onset of acute and chronic degradative events. These include blood brain barrier disruption, edema, demyelination, axonal damage and neuronal cell death. Posttraumatic inflammation after spinal cord injury has been implicated in the secondary injury that ultimately leads to neurologic dysfunction. Studies after spinal cord contusion have shown expression of several chemokines early after injury and suggested a role for them in the ordered recruitment of inflammatory cells at the lesion site (McTigue et al. [1998] J. Neurosci. Res. 53:368-376; Lee et al., [2000] Neurochem Int). We have demonstrated previously that infusion of the broad-spectrum chemokine receptor antagonist (vMIPII) in the contused spinal cord initially attenuates leukocyte infiltration, suppresses' gliotic reaction and reduces neuronal damage after injury. These changes are accompanied by increased expression of bcl-2, the endogenous apoptosis inhibitor, and reduced neuronal apoptosis (Ghirnikar et al. [2000] J. Neurosci. Res. 59:63-73). We demonstrate that 2 and 4 weeks of vMIPII infusion in the contusion-injured spinal cord also results in decreased hematogenous infiltration and is accompanied by reduced axonal degeneration in the gray matter. Luxol fast blue and MBP immunoreactivity indicated reduced myelin breakdown in the dorsal and ventral funiculi. Increased neuronal survival in the ventral horns of vMIPII infused cords was seen along with increased bcl-2 staining in them. Immunohistochemical identification of fiber phenotypes showed increased presence of calcitonin gene related peptide, choline acetyl transferase and tyrosine hydroxylase positive fibers as well as increased GAP43 staining in treated cords. These results suggest that sustained reduction in posttraumatic cellular infiltration is beneficial for tissue survival. A preliminary report of this study has been published (Eng et al. [2000] J. Neurochem. 74(Suppl):S67B). In contrast to vMIPII, infusion of MCP-1 (9 76), a N-terminal analog of the MCP-1 chemokine showed only a modest reduction in cellular infiltration at 14 and 21 dpi without significant tissue survival after spinal cord contusion injury. Comparing data on tissue survival obtained with vMIPII and MCP-1 (9-76) further validate the importance of the use of broad spectrum antagonists in the treatment of spinal cord injury. Controlling the inflammatory reaction and providing a growth permissive environment would enhance regeneration and ultimately lead to neurological recovery after spinal cord injury. J. Neurosci. Res. 64:582-589, 2001. Published 2001 Wiley-Liss, Inc. PMID- 11398182 TI - mDll1 and mDll3 expression in the developing mouse brain: role in the establishment of the early cortex. AB - The Delta/Notch signalling system is involved in several developmental processes. During fly neurogenesis, Delta expression defines the fate of neuronal precursors and inhibits neighboring Notch-expressing cells from acquiring a neural fate, a process known as lateral inhibition. In vertebrates, recent evidence demonstrates that Notch activation can positively determine cell fate and affect neuronal process extension. Nevertheless, Delta-like expression patterns during brain development are relatively unknown. Using a transgenic mouse, which expresses LacZ under the mDll1 promoter, we show by immunofluorescence that in the developing telencephalon mDll1 is expressed in undifferentiated cells in close contact with radial glial cells. Based on in situ hybridization data on mDll1 and mDll3 mRNA expression and on the immunohistochemical detection of beta galactosidase in the Dll1-lacZ transgenic mouse, we suggest that mDll1 and mDll3 are involved in the establishment of the early cortical plate and that mDll1 expressing cells are in close contact with radial glial cells, thereby modulating the latter population, which is known to express Notch1. Furthermore, we suggest that the decrease in mDll1 mRNA found toward the end of gestation could be related, first, to the slowing of neurogenesis and, second, to the differentiation of the radial glial cell population into astrocytes. PMID- 11398183 TI - Decreases in calmodulin binding proteins and calmodulin dependent protein phosphorylation in the medial preoptic area at the onset of maternal behavior in the rat. AB - The onset of maternal behavior is characterized by the action of certain hormones, neuropeptides and neurotransmitters and a concomitant increase in the expression of c-Fos in the medial preoptic area (MPOA) but the signaling events that lie between have not been characterized. Because several of these hormones, neuropeptides and neurotransmitters function by activating Ca(2+)/calmodulin (CaM) mediated signaling pathways, many of which can lead to c-Fos expression, the goal of the current work was to identify calmodulin binding proteins (CaMBPs) or specific CaM-dependent phosphoproteins that might be involved. Probing of SDS PAGE gels of extracts from the hippocampus, parietal cortex, basolateral amygdala and MPOA with recombinant (35)S-VU1-calmodulin (CaM) revealed 30 Ca(2+)-dependent and 4-6 Ca(2+)-independent CaMBPs. Statistically significant maternal behavior related decreases in four Ca(2+)-dependent CaMBPs ( approximately 31 kDa, 50% decrease; approximately 33 kDa, 32%; approximately 50 kDa, 35%; approximately 60 kDa, 33%) were observed specifically in the MPOA. Numerous proteins were phosphorylated in a Ca(2+) CaM-dependent manner with two (MWs approximately 61 Da, approximately 58 kDa) showing a lack of phosphophorylation only in the MPOA. The selective decrease in CaMBPs coupled with the absence of CaM-dependent phosphoproteins implies that changes in Ca(2+)/CaM-mediated signaling may mediate some of the MPOA-specific processes during the onset of maternal behavior in the rat. PMID- 11398184 TI - Dopamine receptor antagonists prevent the d-amphetamine-induced increase in calcitonin gene-related peptide levels in ventral striatum. AB - Microdialysis in conjunction with radioimmunoassay (RIA) were used to study the effects of acute d-amphetamine or dopamine (DA) receptor antagonists administration on extracellular concentrations of calcitonin gene-related peptide (CGRP) in the ventral striatum of the rat. One hour after the subcutaneous (s.c.) injection of saline, the DA-D(1) receptor antagonist SCH 23390 (0.3 mg/kg) or the DA-D(2/3) receptor antagonist raclopride (1.0 mg/kg), one additional s.c. injection of saline or d-amphetamine (1.5 mg/kg) was given. The dialysates were collected at 60-min intervals; CGRP-like immunoreactivities (-LI) were determined by RIA. d-Amphetamine significantly increased extracellular CGRP-LI concentrations compared to the control animals. Administration of either SCH 23390 or raclopride did not significantly affect CGRP-LI concentrations. Pretreatment with either SCH 23390 or raclopride abolished the stimulatory effect of d-amphetamine on CGRP-LI levels. The results show that d-amphetamine administration results in an increase in extracellular concentrations of CGRP in the ventral striatum through a mechanism that appears to involve stimulation of either DA-D(1) or DA-D(2/3) receptors. The results also indicate that changes in dopaminergic neurotransmission affect CGRP outflow in the ventral striatum in a phasic but not tonic manner. PMID- 11398185 TI - Constitutive expression and delayed light response of casein kinase Iepsilon and Idelta mRNAs in the mouse suprachiasmatic nucleus. AB - Casein kinase Iepsilon (CKIepsilon) and casein kinase Idelta (CKIdelta) phosphorylate clock oscillating mPER proteins, and play a key role in the transcription (post)translation feedback loop that generates circadian rhythm. In the present study, the expression profiles of CKIepsilon and CKIdelta mRNAs were examined in the mice clock center, suprachiasmatic nucleus (SCN). Moderate levels of CKIepsilon and CKIdelta mRNAs were constantly expressed in the SCN in both light:dark and constant dark conditions. This finding supports the hypothesis that CKI may form a constant threshold to the nuclear entry of mPER proteins as in the Drosophila homologue, double-time. Further, we demonstrated that the light exposure at subjective night induced a delayed increase in CKIepsilon and CKIdelta mRNAs in the SCN. CKIepsilon and CKIdelta proteins may play a role on light-induced phase-shift. PMID- 11398186 TI - GABA(A) receptor gene expression in rat cortex: differential effects of two chronic diazepam treatment regimes. AB - Diazepam is widely prescribed as an anxiolytic but its therapeutic application is limited because with daily use tolerance develops to certain aspects of its pharmacological profile. We compared the effects of two dosing paradigms on GABA(A) receptor gene expression and benzodiazepine binding characteristics. Equivalent daily doses of 15 mg/kg/day diazepam were delivered either via constant infusion or daily subcutaneous injection for 14 days. The two distinct treatment regimes produced significantly different changes in GABA(A) receptor alpha4-, beta2-, beta3- and gamma1-subunit mRNA steady-state levels. Similar changes in the GABA enhancement of flunitrazepam binding and the BZ3/BZ2 subtype ratio determined ex vivo were produced, however, significant differences were found in [(3)H]-Ro 15-4513 binding between cortical tissue from diazepam injected animals compared with diazepam infused animals. Our data suggest that it is the diurnal fluctuations in receptor occupancy that are responsible for the different effects produced by these two dosing regimes. PMID- 11398187 TI - Histochemical study of acute and chronic intraperitoneal nicotine effects on several glycolytic and Krebs cycle dehydrogenase activities in the frontoparietal cortex and subcortical nuclei of the rat brain. AB - The effects of nicotine on the activity of different dehydrogenases in frontoparietal regions and subcortical nuclei of the rat brain have been studied using histochemical methods. Nicotine sulphate was intraperitoneally administered in acute (4 mg/kg/day x 3 days) or chronic (ALZET osmotic pump providing 2 mg/kg/day x 15 days) doses. The enzymes analyzed were glyceraldehyde-3-phosphate, lactate, malate and succinate dehydrogenases (gly3PDH, LDH, MDH, and SDH, respectively). The results demonstrate that chronic as well as acute administration of nicotine produced strong increases in all these enzymatic activities in the superior layers (I, II and III) of the frontoparietal cortex (cingulate, motor and somatosensory regions); but high increases were not seen in the deeper layers of the cortex or in the subcortical nuclei (substantia nigra, caudate-putamen, nucleus accumbens or nucleus basalis magnocellularis). These hyperactivities were produced in brain regions with normally low enzymatic activity (cortex), but not in those with great intensity (subcortical nuclei). The results are in rough agreement with previous reports on nicotine-induced increases in glucose utilization, gly3PDH genic expression and neuronal hyperactivity in the brain cortex; but significant discrepancies between the cortical enzymatic maps and those obtained both in these studies and others on nicotine(N)-receptor localization have been appreciated. The results support the hypothesis that nicotinic cholinergic drugs can have metabolic, long-lasting stimulant effects on cortical neurons at specific points (probably layer III pyramidal cells and structures with alpha7-N-receptors) of the cortical circuits that could be of great interest in improving altered cognitive functions that are present in Alzheimer disease, as well as in other less severe mental disturbances. Mitochondrial hyperfunction should also be evaluated as a possible side-effect (as an oxidative stress inductor) of these kinds of drugs. PMID- 11398188 TI - TrkA induces differentiation but not apoptosis in C6-2B glioma cells. AB - Nerve growth factor (NGF) binds to the TrkA tyrosine kinase and the p75 neurotrophin receptors. Depending upon which receptor is activated, NGF can induce differentiation or apoptosis. C6-2B glioma cells express the p75 receptor, but NGF decreases their growth only when TrkA is introduced (C6trk). It is unclear, however, whether TrkA reduces C6-2B cell growth by apoptosis or differentiation. To examine which mechanisms account for the anti-proliferative effect of NGF in these cells, we first analyzed whether NGF causes apoptosis by flow cytometry, two-site immunoassay and in situ TUNEL. None of these methods indicated that C6trk undergo apoptosis. Additional apoptotic markers, such as Bcl 2, Bax, Bad, p53, caspase 3, and NF-kappaB were also used. C6trk cells exhibited lower levels of Bcl-2 compared with the parental C6 mock cells, but no changes in the levels of other apoptotic proteins. Moreover, NGF increased AP-1 binding activity in C6trk cells, suggesting that NGF may induce differentiation. We then examined whether TrkA changes the glioma phenotype. In C6trk cells, but not in C6mock cells, NGF enhanced the levels of neuron-specific enolase as well as the levels of A2B5 and 2', 3'-cyclic nucleotide 3'-phosphodiesterase, markers for oligodendrocytes, without affecting the expression of other neuronal markers. Our data suggest that the antiproliferative properties of TrkA may rely on its ability to induce differentiation of C6 cells from undifferentiated glioma to oligodendrocytes. PMID- 11398189 TI - Cobalt chloride induces PC12 cells apoptosis through reactive oxygen species and accompanied by AP-1 activation. AB - Reactive oxygen species (ROS) are supposed to play an important role in hypoxia- and ischemia/reperfusion-mediated neuronal injury with the characteristics of apoptosis. There are many reports showing that cobalt chloride (CoCl(2)) could mimic the hypoxic responses in some aspects including production of ROS in cultured cells. The cytotoxicity of CoCl(2) and its molecular mechanisms have yet to be elucidated. We report that CoCl(2) triggered neuronal PC12 cells apoptosis in a dose- and time-dependent manner. Apoptosis was demonstrated by morphological changes and DNA fragmentation, and was dependent on macromolecular synthesis. Apoptosis was also confirmed by the decrease of the expression of Bcl-X(L). To our knowledge, this is the first documentation of the apoptotic induction of CoCl(2) on PC12 cells. Furthermore, ROS production in PC12 cells was increased during CoCl(2) treatment. Antioxidants, which could inhibit ROS production, significantly blocked CoCl(2)-induced apoptosis, suggesting that apoptosis is mediated by ROS production. We also observed a significant increase of the DNA binding activity of AP-1 in response to CoCl(2) and this increase was blocked by antioxidants, showing that CoCl(2)-induced apoptosis is accompanied by ROS activated AP-1. CoCl(2)-treated PC12 cells may serve as an in vitro model for studies of molecular mechanisms in ROS-linked neuronal disorders. PMID- 11398190 TI - Mechanisms of cell death in primary cortical neurons and PC12 cells. AB - Increasing evidence suggests that the regulation of neuronal cell death is complex. In this study we compared the neurotoxic effects of tumor necrosis factor-alpha (TNFalpha), nitric oxide, and thrombin on primary rat cortical cell cultures and the neuronal PC12 cell line. Release of lactate dehydrogenase (LDH) and the intracellular accumulation of nucleosomes were used as indicators of necrosis and apoptosis, respectively. There was significant LDH release in both neuronal cell types, however, the pattern of LDH release was variable and agonist dependent. In response to the nitric oxide generator, sodium nitroprusside (SNP), cortical cells exhibited pronounced LDH release and dramatic morphologic changes, whereas in differentiated PC12 cells, TNFalpha evoked release of LDH with no associated morphologic changes. Both neuronal cell types, but not undifferentiated PC12 cells, responded to TNFalpha and thrombin with increased apoptosis. Caspase inhibition, but not antioxidant treatment, reduced nucleosome accumulation in primary cortical cells, but not in differentiated PC12 cells. In the differentiated PC12 cells, caspase inhibition reduced TNFalpha-mediated LDH release, but not nucleosome accumulation. These data suggest mechanisms involved in neuronal cell death utilize multiple pathways that vary depending on the neurotoxic insult and are also influenced by subtle differences among neuronal cell phenotypes. PMID- 11398191 TI - Plasma membrane calcium ATPase plays a role in reducing Ca(2+)-mediated cytotoxicity in PC12 cells. AB - In many cell types, cell death induced by a variety of insults is accompanied by an increase in intracellular calcium. The Ca(2+) homeostatic mechanisms affected by such insults, however, have not been fully determined. Recent evidence indicates that kainic acid-induced seizures alter plasma membrane calcium ATPase mRNA expression within vulnerable hippocampal cell populations before the onset of cell death. We examined the effects of altering plasma membrane calcium ATPase expression on cell vulnerability in rat pheochromocytoma 12 cells. Pheochromocytoma 12 cells are vulnerable to Ca(2+) overload induced by the Ca(2+) ionophore A23187. Reverse transcriptase-PCR and Western blot data indicated that plasma membrane calcium ATPase isoform 4b constitutes a major calcium pump isoform in the pheochromocytoma 12 cells. Therefore, permanently transfected pheochromocytoma 12-derived cell lines were established that either over expressed plasma membrane calcium ATPase isoform 4b, or suppressed the expression of the endogenous plasma membrane calcium ATPase isoform 4. Over-expressing clones were less vulnerable to Ca(2+)-mediated cell death induced by A23187 whereas "antisense" clones were considerably more susceptible. These data indicate that regulation of plasma membrane calcium ATPase expression may be critical to cellular survival when cells are exposed to pathological increases in intracellular calcium. PMID- 11398193 TI - Involvement of transcription factor HNF3gamma in the effect of o-aminoazotoluene on glucocorticoid induction of tyrosine aminotransferase in mice sensitive to its hepatocarcinogenic action. AB - In the rodent liver, hepatocarcinogens inhibit the glucocorticoid induction of several liver-specific genes, including tyrosine aminotransferase (TAT). A distinct positive correlation exists in mice between the extent of inhibition of TAT induction after acute administration of o-aminoazotoluene (OAT) and the frequency of liver tumors after chronic exposure to the carcinogen. To elucidate the mechanism of the carcinogenic action, the effects of OAT on the DNA-binding activity of several transcription factors participating in the glucocorticoid regulation of TAT gene expression were studied. The experimental inbred male mice were sensitive (A/He and SWR/J, tumor induction frequency of 75-100%, TAT induction inhibition of 35-50%) and resistant (CC57BR/Mv and AKR/J, 0-6% and 10 15%, respectively) to OAT. Gel retardation experiments showed that hepatocyte nuclear factor 3 (HNF3)gamma DNA-binding activity was strongly reduced in nuclear extracts from the livers of OAT-treated A/He and SWR/J mice but only slightly reduced in CC57Br/Mv and AKR/J mice. The DNA-binding activities of Ets, AP1 family members, and GME binding proteins were unaffected. HNF3gamma DNA-binding activity was reduced by 1 h after OAT administration and remained low for 1 mo, as did inhibition of TAT induction in the liver. These results suggested that the inhibitory effect of OAT on the glucocorticoid induction of TAT is mediated by reduced HNF3gamma DNA-binding activity. PMID- 11398192 TI - Epigenetic silencing of PEG3 gene expression in human glioma cell lines. AB - Genomic imprinting, the phenomenon in which alleles of genes are expressed differentially depending on their parental origins, has important consequences for mammalian development, and disturbance of normal imprinting leads to abnormal embryogenesis and some inherited diseases and is also associated with various cancers. In the context of screening for novel imprinted genes on human chromosome 19q13.4 with mouse A9 hybrids, we identified a maternal allele specific methylated CpG island in exon 1 of paternally expressed imprinted gene 3 (PEG3), a gene that exhibits paternal allele-specific expression. Because PEG3 expression is downregulated in some gliomas and glioma cell lines, despite high level expression in normal brain tissues, we investigated whether the loss of PEG3 expression is related to epigenetic modifications involving DNA methylation. We found monoallelic expression of PEG3 in all normal brain tissues examined and five of nine glioma cell lines that had both unmethylated and methylated alleles; the remaining four glioma cell lines exhibited gain of imprinting with hypermethylated alleles. In addition, treatment of glioma cell lines with the DNA demethylating agent 5-aza-2'-deoxycytidine reversed the silencing of PEG3 biallelically. In this article, we report that the epigenetic silencing of PEG3 expression in glioma cell lines depends on aberrant DNA methylation of an exonic CpG island, suggesting that PEG3 contributes to glioma carcinogenesis in certain cases. PMID- 11398194 TI - Characterization of the CDKN2A and ARF genes in UV-induced melanocytic hyperplasias and melanomas of an opossum (Monodelphis domestica). AB - We examined the involvement of the cyclin-dependent kinase inhibitor 2A (CDKN2A) locus in the pathogenesis of ultraviolet (UV) radiation-induced melanomas in an opossum (Monodelphis domestica) melanoma model in which suckling young were exposed to UVB to produce melanocytic lesions. Monodelphis CDKN2A and alternated reading frame (ARF) cDNAs were cloned and sequenced, and the expression patterns of these genes were determined by reverse transcription-polymerase chain reaction in normal tissues, 39 primary melanocytic skin lesions, and two tumor-derived cell lines, one nonmetastatic and one metastatic. Primary melanocytic lesions, including hyperplasias, benign melanomas, melanomas metastatic to lymph nodes, and melanomas metastatic to nodes and additional visceral organs, were categorized accordingly as types I-IV. Levels of CDKN2A transcripts were most abundant in type III tumor samples and the metastatic cell line but absent in the nonmetastatic cell line. ARF transcripts were expressed in all tumors and cell lines. A UV-signature mutation was detected with the wild-type allele at the CDKN2A locus in type II and III primary tumor samples and in the nonmetastatic cell line. Interestingly, in the metastatic cell line, only the mutant allele was present and expressed. These data suggest dynamic changes in the expression and/or structure of the CDKN2A and ARF genes represent one molecular defect associated with the etiology of melanoma formation and progression in the Monodelphis model system. PMID- 11398195 TI - Inhibition of human papillomavirus-16 long control region activity by interferon gamma overcome by p300 overexpression. AB - Although interferons (IFNs) are currently used in the treatment of various human papillomavirus (HPV)-associated lesions, their mechanisms of action are still unclear. In this study, we clearly demonstrated that IFN-gamma was a strong inhibitor of HPV-16 long control region (LCR) activity in two human cervical carcinoma cell lines. The effect of IFN-gamma was dose dependent. We investigated whether the effect of IFN-gamma on HPV-16 LCR could involve the inhibition of the CREB-binding protein (CBP)/p300 family of transcriptional coactivators. In support of this model, we demonstrated by transfection experiments that a 12S E1A mutant (RG2), which interacts poorly with p300 and CBP in comparison to wild-type E1A, was less able to repress human papillomavirus (HPV) 16 long control region (LCR) than wild-type E1A. More important, overexpression of p300 was able to increase the HPV-16 LCR activity and to overcome inhibition by IFN-gamma. Finally, we demonstrated that p300 could cooperate with c-jun to activate HPV-16 LCR. According to our results, IFN-gamma might inhibit HPV-16 LCR transcription by activating the signal transducer and activator of transcription 1alpha, which in turn might compete for p300/CBP binding with specific transcription factors involved in LCR activation. PMID- 11398197 TI - Molecular characterization of the loss of p75(NTR) expression in human prostate tumor cells. AB - The low-affinity nerve growth factor receptor p75(NTR) is a 75-kDa glycoprotein that belongs to the tumor necrosis factor receptor superfamily and has been implicated in the induction of apoptosis in various tissues and cell lines. Immunohistochemistry on tissue sections from radical prostatectomies has shown that expression of p75(NTR) is limited to the epithelial cells. Western blot and immunohistochemical analyses have also shown a progressive loss of p75(NTR) expression in prostate epithelial cells during the malignant progression of organ confined adenocarcinomas, with complete loss of expression in the naturally occurring prostate tumor cell lines DU-145, PC-3, LNCaP, and TSU-pr1, which were derived from metastases. Reintroduction of p75(NTR) expression into the TSU-pr1 tumor cell line was shown to reestablish the ability of these cells to undergo p75(NTR)-mediated apoptosis. It is not known whether this loss of expression is due to deletion of part or the entire p75(NTR) gene or to other factors. Through the use of southern blotting and polymerase chain reaction (PCR), we showed that loss of p75(NTR) protein expression was not due to deletion or loss of the gene. Furthermore, through reverse transcription-PCR, RNase protection, and the chromatin immunoprecipitation assay, we showed that transcription of the p75(NTR) gene occurred in these prostate tumor cell lines. Finally, through transient transfection using two constructs of p75(NTR), one containing the full 2-kb 3' untranslated region and one that contains only a few hundred bases of the 3' untranslated region (UTR), we showed that the 3' UTR may have a role in the loss of p75(NTR) expression in prostate cancer. PMID- 11398196 TI - Transformation of kidney epithelial cells by a quinol thioether via inactivation of the tuberous sclerosis-2 tumor suppressor gene. AB - Although hydroquinone (HQ) is a rodent carcinogen, because of its lack of mutagenicity in standard bacterial mutagenicity assays it is generally considered a nongenotoxic carcinogen. 2,3,5-Tris-(glutathion-S-yl)HQ (TGHQ) is a potent nephrotoxic metabolite of HQ that may play an important role in HQ-mediated nephrocarcinogenicity. TGHQ mediates cell injury by generating reactive oxygen species and covalently binding to tissue macromolecules. We determined the ability of HQ and TGHQ to induce cell transformation in primary renal epithelial cells derived from the Eker rat. Eker rats possess a germline inactivation of one allele of the tuberous sclerosis-2 (Tsc-2) tumor suppressor gene that predisposes the animals to renal cell carcinoma. Treatment of primary Eker rat renal epithelial cells with HQ (25 and 50 microM) or TGHQ (100 and 300 microM) induced 2- to 4-fold and 6- to 20-fold increases in cell transformation, respectively. Subsequently, three cell lines (The QT-RRE 1, 2, and 3) were established from TGHQ-induced transformed colonies. The QT-RRE cell lines exhibited a broad range of numerical cytogenetic alterations, loss of heterozygosity at the Tsc-2 gene locus, and loss of expression of tuberin, the protein encoded by the Tsc-2 gene. Only heterozygous (Tsc-2(EK/+)) kidney epithelial cells were susceptible to transformation by HQ and TGHQ, as wild-type cells (Tsc-2(+/+)) showed no increase in transformation frequency over background levels following chemical exposure. These data indicate that TGHQ and HQ are capable of directly transforming rat renal epithelial cells and that the Tsc-2 tumor suppressor gene is an important target of TGHQ-mediated renal epithelial cell transformation. PMID- 11398198 TI - Wingless-type frizzled protein receptor signaling and its putative role in human colon cancer. AB - We wish to identify new candidate genes involved in the pathogenesis of human colon cancer to better understand the diversity of phenotype presentation that varies from individual to individual. Our working hypothesis is that genetic polymorphism of genes in the Wingless-type (Wnt) frizzled protein receptor pathway is associated with the susceptibility to develop colon cancer. The putative role of the Wnt pathway in sporadic human malignancy of the colon suggests involvement in inherited cancer as well. beta-catenin is the crucial messenger in frizzled receptor signaling, transmitting Wnt-ligand signals such as signals from secreted apoptosis-related proteins to the nucleus. It functions as a genome denunciator by initiating amplification of oncogenes. The net effect of beta-catenin depends on the magnitude of its accumulation in the cytoplasm and, therefore, upon expression profiles of genes in the Wnt pathway. We propose that variations in allelic frequencies of genes involved in the beta-catenin cascade may either promote or impede malignant transformation of the colon. If certain polymorphisms in Wnt signaling through beta-catenin predispose to colon cancer, this might manifest as decreased binding affinity of proteins such as axin or the adenomatous polyposis coli protein to beta-catenin. Association studies are proposed to test the hypothesis, which could serve as an initial step toward understanding the complexity of tumor biology. The clinical rationale in unraveling the genetic susceptibility to cancer lies in identification of a subgroup of individuals who may benefit from beta-catenin targeting agents, which could potentially overcome this genetic instability. PMID- 11398199 TI - Diagnosis and staging of gastrointestinal neoplasms. PMID- 11398200 TI - Role of endoscopy in the staging of esophageal and gastric cancer. AB - Progress in instrumentation and clinical research continues to expand the potential utility of endoscopic ultrasound (EUS) in the treatment of esophageal and gastric cancer. This review focuses on the role of EUS in the staging and treatment planning of patients with esophageal and gastric cancer. PMID- 11398201 TI - Role of endoscopy in staging colorectal cancer. AB - The treatment of colorectal cancer depends in large measure on the depth of tumor invasion and the extent of lymph node involvement. Endoscopic ultrasonography (EUS) has added a new dimension to the evaluation of tumor invasion and lymph node involvement in gastrointestinal cancer. The overall EUS accuracy for colorectal cancer T-staging is 78%, specificity is 73%, and sensitivity is 94%. In determining the nodal involvement by tumor, EUS has an accuracy of 75%, specificity of 73%, and sensitivity of 74%. Comparison with computerized tomography (CT), magnetic resonance imaging (MRI), and MRI with endorectal coil (MRIEC) shows that EUS is an effective single modality for assessing tumor penetration of the rectal wall. It does not, however, allow the assessment of distant metastatic disease. For assessing lymph node involvement, MRIEC offers the most comprehensive information. PMID- 11398202 TI - Role of ultrasonography in the staging of gastrointestinal neoplasms. AB - This article reviews the capabilities and limits of ultrasonography (US) in the staging of gastrointestinal neoplasms. US is a well-established tool in the investigation of abdominal diseases. Its role is very important in the first approach to liver, gallbladder, biliary, and pancreatic diseases, but its abilities for accurate staging may be limited by various factors, which will be discussed. In the evaluation of the stomach and intestine, US is rarely utilized, but it can occasionally demonstrate an unsuspected gastrointestinal mass that usually must be evaluated further with specific techniques (endoscopy and barium studies) to confirm the diagnosis and to perform an accurate staging (with endosonography and computed tomography). PMID- 11398203 TI - Role of conventional radiology in the diagnosis and staging of gastrointestinal tract neoplasms. AB - Preoperative diagnosis and staging of primary gastrointestinal (GI) neoplasms are challenges for both clinicians and radiologists. Barium studies are very sensitive in disclosing primary malignancies, even at an early stage. Radiologic signs depend on the evolutional stage of the disease and its appearance on gross pathology. A neoplasm may be manifested on barium studies by a wide spectrum of findings, including mucosal abnormalities, mass presence, ulcerative lesions, submucosal infiltration, and lumen stenosis. Advanced disease can be accurately diagnosed, whereas early cancer should be differentiated from other neoplastic or inflammatory disorders by meticulous analysis of radiologic findings. The extent of GI involvement and multiplicity of the lesions can be assessed on barium studies. In the staging of GI neoplasms, barium studies are of little value. Skeletal survey by conventional radiographs and chest plain films can reveal distant metastases in a short time and with low cost, although they are not that sensitive to the detection of early or subtle lesions. The exact role of conventional radiology in the imaging workup of GI malignancies depends on local expertise and availability of other diagnostic techniques and modalities. PMID- 11398204 TI - Role of computerized tomography in the staging of gastrointestinal neoplasms. AB - Gastrointestinal neoplasms are very common diseases, and the first challenge for clinicians is to define the extent of the tumor in order to plan the best treatment. The role of computerized tomography in assessing this kind of patient is well established worldwide. This article reviews the capabilities and the limits of this imaging technique in the staging of the tumors of the digestive tract (liver, gallbladder, biliary tract, pancreas, esophagus, stomach, small bowel, and colon). PMID- 11398205 TI - Role of magnetic resonance imaging in the staging of gastrointestinal neoplasms. AB - A concise review is presented of the current applications and techniques of magnetic resonance imaging (MRI) in the field of diagnostic imaging of oncologic disease of the gastrointestinal tract, with a summary of the diagnostic possibilities of MRI in the various types of tumor pathology of the gastrointestinal tract. We conclude with a review of the specific situations in which MRI presents distinct advantages over other diagnostic imaging methods (such as computed tomography and ultrasound). PMID- 11398206 TI - Role of angiographic techniques in the preoperative staging and management of gastrointestinal neoplasms. AB - Gastrointestinal neoplasms are very common diseases, and their management does not usually require angiography for diagnostic and staging purposes. However, angiography may be required for further refinements in staging of vascular involvement or to obtain a detailed preoperative anatomy of the vessels. Finally, angiographic techniques may be useful for palliative or preoperative locoregional chemotherapy, and to treat hemorrhagic complications. This article reviews the capabilities and limits of angiographic techniques in the assessment and management of tumors of the alimentary tract. PMID- 11398208 TI - Diagnostic and prognostic tumor markers in the gastrointestinal tract. AB - The gastrointestinal tract is the most common site of malignancies of any anatomic system in the body. An early detection of primary tumors of the bowel, pancreas, liver, stomach, and esophagus is often difficult in asymptomatic patients and for this reason these tumors are often detected at a relatively advanced stage, when symptoms lead to a diagnostic evaluation. Furthermore, gastrointestinal tract tumors have an extremely variable prognosis; thus, the identification of new prognostic parameters may be useful for selecting patients to more tailored therapies. In this work, the main molecular, genetic, tissular, and circulating tumor markers proposed for diagnosis and prognosis of gastrointestinal malignancies are reviewed and discussed. PMID- 11398207 TI - Positron emission tomography (PET) and other nuclear medicine modalities in staging gastrointestinal cancer. AB - Gastrointestinal (GI) cancers account for the second highest number of new tumor cases and deaths per year in the United States and Western Europe. The most frequently involved sites are, in descending order, the colorectum, stomach, pancreas, liver, bile duct, and esophagus. The most common tumor type is adenocarcinoma. Among the epithelial cancers, great attention has recently been given to the tumors of neuroendocrine origin. These concepts are relevant because nuclear medicine imaging is based on visualization by means of a particular uptake of radiolabelled tracers in cancer cells that concentrate the radioactive signal. This signal is detected and reconstructed in planar or tomographic images. Different radiopharmaceuticals have been proposed for diagnostic application in oncology (such as radiolabelled monoclonal antibodies (MAbs), receptor tracers, and positron-emitting radiopharmaceuticals), and they are currently used as tracers for tumor detection with different modalities and techniques. Most of these techniques demonstrate their clinical usefulness in the diagnosis of GI cancer. This work is not intended to be a comprehensive review of all the extensive experience and possibilities of nuclear medicine for the diagnosis of GI tumors; rather, it aims to summarize the current status of the most important approaches and their main indications in staging GI cancers. PMID- 11398210 TI - Spinal stenosis. Severity determines treatment plan. PMID- 11398209 TI - Role of video-laparoscopy in the staging of intra-abdominal lymphomas and gastrointestinal cancer. AB - There is good category II/III evidence that video-laparoscopic staging is valuable in certain gastrointestinal (gastric, esophageal, pancreatic, and hepatobiliary) and intra-abdominal lymphomas, but no category I evidence (based on prospective randomized trials). The evidence available is all retrospective, but of sufficient consistency to indicate that laparoscopic staging adds to the primary (imaging) staging and often alters the clinical stage of the disease and hence the management of the individual patient. The advent of laparoscopic contact ultrasound (LCU) scanning has improved the staging accuracy for pancreatic and hepatobiliary neoplasms. The laparoscopic approach also offers a means of surgical palliation in certain patient groups. However, there are a number of unresolved issues concerning the use of video-laparoscopy. The most important concerns whether staging laparoscopy should be performed immediately before scheduled surgery or as a separate intervention. The cost-efficacy of these two management options needs to be evaluated in prospective studies. In some centres, laparoscopic staging is being conducted by gastroenterologists and hepatologists. This raises issues of safety and ability to undertake certain procedures that may be necessary during the laparoscopic staging. PMID- 11398211 TI - Health tips. Adding pounds when you're underweight. PMID- 11398212 TI - Second thoughts on a common treatment for COPD. PMID- 11398213 TI - Researchers study medical cause of the yips. PMID- 11398214 TI - Postsurgical recovery. What to expect. PMID- 11398215 TI - Mad cow disease. Behind the headlines. PMID- 11398216 TI - Nutrition quiz. Test your knowledge of nutrition information. PMID- 11398217 TI - I'm just getting over a really bad cold. Would it help if I tossed out my old toothbrush and got a new one to avoid reinfecting myself with any cold virus that may be on the brush? PMID- 11398218 TI - My grandson dropped a glass thermometer on the kitchen floor. It shattered and spilled the mercury. What health risk does the mercury pose? How should I have cleaned it up? PMID- 11398219 TI - Breast cancer. Early detection and prompt treatment are critical. PMID- 11398220 TI - Legal regulation of cancer surveillance: Canadian and international perspectives. PMID- 11398221 TI - The contested lessons of euthanasia in The Netherlands. PMID- 11398222 TI - Children living with HIV: reshaping law and policy in Quebec to preserve and promote their rights. PMID- 11398223 TI - Home care in Ontario: the case for copayments. PMID- 11398224 TI - Telling the truth? Disclosure, therapeutic privilege and intersexuality in children. PMID- 11398225 TI - Constitutional jurisdiction over health in Canada. PMID- 11398226 TI - Coping with stress in the workplace: implications for new health professionals. AB - Because health care professionals must provide high-quality care while simultaneously adjusting to the new demands of a rapidly changing health care system, work stress and possibly even burnout are prevalent issues for both senior professionals and recent health science graduates upon entry into the workforce. This article examines the experience of stress and the importance of coping abilities, particularly focusing on students enrolled in health professional courses and those who have recently entered the workforce. It also presents research pertaining to whether newly graduated health professionals are coping effectively with the demands of work. Suggestions for improving stress coping abilities are given. PMID- 11398227 TI - Do learning style and learning environment affect learning outcome? AB - This study compared learning outcomes of students with different learning styles, as identified by the Kolb Learning Style Inventory indicators, in a traditional in-class environment with those taking the same course via distance education. The above-average scores were evenly distributed, 47% of the in-class group and 43% of the distance group. For three of the four learning styles, there was no relationship to learning outcome or environment. The Diverger group did show a relationship with above-average scores in the distance group (83%). The findings support that the classroom or distance environment did not influence learning outcome. Learning style did not appear to affect learning outcome in either group, except that the Diverger learning style may have a positive relationship to learning in the distance environment. PMID- 11398228 TI - Allied health representation: a call for action. AB - The Coalition of Allied Health Leadership (CAHL) 2000 Representation Team sought 1) to secure Federal advisory appointments for allied health professionals and 2) to connect allied health representatives with Federal advisory committees, councils, boards, and other deliberative bodies. Among the deliberative bodies providing recommendations on a broad range of issues to the President of the United States and the Executive Branch, there are over 1,000 advisory committees, councils, and boards, with more than 20,000 members. Recommendations made by the deliberative bodies include those related to health care. The literature and Web sites reveal allied health professionals have little or no representation on the bodies that represent allied health professionals and their constituents. These findings provide insights into Federal-level deliberative bodies to which allied health professionals have access or on which they warrant representation. This article reports background information, including the CAHL 2000 Representation Team objectives; an overview of federal advisory committees; recommendations for gaining access to deliberative bodies and active participation in fulfillment of Healthy People 2010 goals; and continued commitment to such representation by the CAHL and allied health professionals. PMID- 11398229 TI - Incorporating transcultural care education in allied health curricula. AB - A clinical skill recently identified as a necessary curricular component in allied health programs is transculturally competent care. It is imperative to teach allied health students transcultural care skills, first, because the population of the United States is becoming more ethnically, culturally, and racially diverse, and the allied health workforce must be prepared for this change. Second, Western medicine has historically reflected European values and traditions. Because other cultures view health and disease differently, health care workers must understand and be respectful of values and traditions that differ from their own. And finally, organizations that accredit health care facilities and academic programs, and licensing boards, are recognizing cultural competency as an important clinical skill for all health care workers. Based on existing literature on transcultural care education, which primarily comes from nursing, this article suggests a model for its incorporation in allied health curricula. PMID- 11398230 TI - Ergonomics: situated at the crossroads of conflict among the three branches of government. PMID- 11398231 TI - Differences in attitudes regarding teaching and research orientations between allied health faculty and administrators in academic health centers. AB - The congruence between faculty's and administrators' attitudes toward teaching, research, and the rewards associated with them has implications for the practices the groups follow and for the decisions they make related to promotion, tenure, and merit. This article reports the findings from a national study of allied health faculty and administrators at public academic health centers regarding their attitudes toward teaching, research, and the reward systems related to both. The administrators and faculty at the participating academic health centers did not differ in their views regarding research orientation and rewards related to research. The faculty, including tenured and tenure-track, and department chairs indicated a higher teaching orientation than did the deans, while tenure track faculty expressed less belief that rewards influence teaching than did tenured faculty, department chairs, and deans. PMID- 11398232 TI - Determinants of job satisfaction among selected care providers in Kuwait. AB - The study reports job satisfaction among a sample of 370 physicians, nurses, pharmacists, and medical laboratory technologists in the Ministry of Health hospitals in Kuwait. Job satisfaction was measured using Dunnette's scale. The respondents in all four categories were satisfied with all aspects of their jobs except salary. The medical laboratory technologists were, however, dissatisfied with professional advancement as well. The physicians were found to be most satisfied, closely followed by nurses and pharmacists, while medical laboratory technologists were least satisfied. Multivariate analysis was used to assess the net effects of background and work environment characteristics on overall job satisfaction. Introduction to job, in-service training, and monthly income had significant positive effects on overall job satisfaction. These findings may have serious implications for health care delivery systems that recruit expatriate care providers. PMID- 11398233 TI - Job choice and personality: a profile of Michigan occupational and physical therapists. AB - The aim of this descriptive study was to compare the personality profiles of occupational therapists (OTs) and physical therapists (PTs) and examine the relationship between personality and job choice. The Kiersey-Bates Personality Inventory (70-item questionnaire) and a brief demographic survey were mailed to a random sample of 400 Ts and 400 PTs in Michigan. A total of 294 completed surveys were returned for analysis, for a combined response rate of 37%. Results showed that while the demographic profiles of OTs and PTs were similar, there were very significant differences between OTs and PTs with respect to personality. The implications of these findings for professional health education are discussed. PMID- 11398234 TI - The influence of personality type on decision making in the physical therapy admission process. AB - The purpose of this study was to identify the personality types of physical therapy (PT) interviewers and applicants, using the Personality Styles (PS) assessment tool, and to determine whether an interview team's personality type influences the rating score given. The PS was validated in a study of 298 students who completed the Myers-Briggs Type Indicator (MBTI) Form G and a PS assessment. By chi-square analysis the PS model appears to be a valid representation of the MBTI (chi 2 = 86.62, p < 0.00001). Subjects for the interview portion of the study were 282 student applicants, 19 faculty, and 47 clinicians from two PT programs. A randomly assigned faculty/clinician team interviewed each applicant. Two one-way ANOVAs were performed with interview score as the dependent variable and 1) applicant personality type in relation to faculty/clinician team (same, different from both, like one) and 2) applicant personality type as the independent variables. Internal consistency of the interview rating form was alpha = 0.89. Mean interview score was 33.97/42 (SD 4.59). Interview scores were not significantly different between applicants who interviewed with clinician/faculty teams that were "like" compared with "not like" the applicants (F0.864; p = 0.423), but were significantly different between applicants with different PS personality types (F3.159; p = 0.026). Although personality type of the interview team did not impact the score given, thereby refuting the presence of interviewer bias, the rating scores did vary according to personality type of the applicant, suggesting a possible stereotyping bias in the criteria used to rate applicants. PMID- 11398235 TI - Characteristics, perceptions, and factors influencing the decision making of physical therapy chairpersons in 2000. AB - The physical therapy (PT) profession is changing. A mailed survey elicited characteristics of current PT chairpersons, such as time spent on responsibilities, influences on decision making, perceptions of working relationships, and satisfaction with decision making, as well as information about their supervisors and the faculty they managed. Chairpersons of 115 (54.28%) accredited PT programs responded. They had been in their positions for a mean of 6.04 years; 63 (57.8%) held doctoral degrees. They had a mean of 25.6 years of experience in PT practice and 17.5 years in academic positions. Their levels of satisfaction and the power and influence they had in their positions were high. However, the time spent on faculty recruitment and mentoring, given the lack of seniority of their faculty, and the small percentage of time they devoted to their own professional development may be concerns. Long-range planning and accreditation duties take up most of the time of PT chairpersons. This information may be helpful to people who seek, or who are recruited for, chairperson positions. It provides a foundational database for longitudinal studies of PT chairpersons. PMID- 11398236 TI - Disaster management for tropical countries. PMID- 11398237 TI - Is immunoproliferative small intestinal disease uncommon in India? AB - Till date only three series of immunoproliferative small intestinal disease (IPSID) describing 22 patients have been reported from India. Seven patients with IPSID in two tertiary referral centers in India are included in the study. Diagnosis was based on typical clinical features [diarrhoea (7/7), weight loss (7/7), clubbing (6/7), fever (3/7), abdominal pain and lump (3/7)], biochemical evidence of malabsorption and duodenal biopsy findings. All patients were young males (mean age 29.8 +/- 11.8 years, range 17-53). Atypical features included gastric involvement (1/7), colonic involvement (1/7) and appearance of pigmented nails following anti-cancer chemotherapy (1/7) which disappeared six months after omitting doxorubin from chemotherapy regimen. Parasitic infestation was common. Ascaris lumbricoides (1/7), Giardia lamblia and hookworm (1/7), Strongyloides stercoralis and Trichuris trichura (1/7). In the latter patient S. stercoralis became disseminated after anti-malignant chemotherapy. One patient had gastric H. pylori infection. Four of the seven patients who were misdiagnosed as tropical sprue were treated with tetracycline. This raises doubt on efficacy of tetracycline alone in treatment of IPSID. One other patient was misdiagnosed and treated as intestinal tuberculosis. Early diagnosis and administration of chemotherapy may improve survival in this disease. PMID- 11398238 TI - Prevalence of hepatitis B virus infection in Chandigarh over a six year period. AB - A total of 6418 samples were received between January 1993 to December 1998 from patients with hepatitis. Blood samples were also collected from 946 apparently healthy subjects. All these samples were tested for Hepatitis B surface antigen (HBsAg) by third generation micro ELISA. The overall HBsAg prevalence rate was 12.8%. The highest prevalence was noted in renal transplant patients (21.7%) followed by patients with acute hepatic disease (15.3%), pregnancy with jaundice (9.4%), chronic renal failure (8.8%), nephrotic syndrome (3.1%), whereas the prevalence rate in control group was 2.4%. The prevalence rate of HBsAg was higher in subjects between 21 to 30 years of age with a male preponderance (Male:Female = 2.8:1). PMID- 11398239 TI - Extrahepatic biliary atresia in Saudi Arabia: the importance of early diagnosis and referral. AB - OBJECTIVE: A retrospective analysis of the medical records of children who were managed for extrahepatic biliary atresia (EHBA) over a 6-year period. MATERIALS AND METHODS: Twelve children were managed at King Fahad National Guard Hospital with a diagnosis of extrahepatic biliary atresia from January 1993 through December 1998. The data obtained included age, sex, clinical presentation, age at referral, investigations, management, complications and follow-up. RESULTS: There were 7 males and 5 females with age of onset of jaundice ranging from 3 days to 42 days (mean 10 days). The mean age at referral was 17 weeks (range 6-49 weeks). Kasai Portoenterostomy (PE) was performed in 7 infants and the mean age at surgery was 8 weeks (range 7 to 10.5 weeks). Four children could not undergo Kasai PE because they were more than 20 weeks old and 1 child although 10 weeks old had extensive liver cirrhosis and was considered not suitable for the procedure. One child who had primary liver transplant outside the Kingdom and 4 children who had Kasai PE are still alive. Seven children are lost to follow-up and are probably dead. CONCLUSION: The incidence of EHBA in Saudi Arabia is unknown and the disease is probably rare based on our review. Although jaundice, pale stools and dark urine were observed in early infancy, referral was always late. Public and health professional's awareness of the clinical features of EHBA is important in early identification of infants and early referral. Starting paediatric hepatobiliary centers in Saudi Arabia will improve the management of these cases. PMID- 11398240 TI - Ultrasonographic evaluation of common duct diameter in pre and post cholecystectomy patients. AB - Thirty four patients with gall bladder disease and normal common duct were investigated prospectively by ultrasonography just before cholecystectomy and at periodic intervals of 1 months, 2 months, 3 months and once in 4-6 months after cholecystectomy. The pre and post cholecystectomy common duct diameters were measured at each visit and compared with each other to determine the statistical significance. No significant change (p > 0.05) occurred in common duct diameter following cholecystectomy. PMID- 11398241 TI - Colonic mucosal changes in portal hypertension. AB - Forty one consecutive patients with portal hypertension (PHT) were evaluated by colonoscopy to study the prevalence, type, extent and predictors of haemorrhoids, colorectal varices, and portal hypertensive colopathy. Specific inquiry and regular follow-up assessed frequency of hematochezia. Twenty five patients with obscure gastrointestinal bleeding without PHT who underwent colonoscopy served as controls. Haemorrhoids were seen in nine of 41 (21.9%) patients with PHT and four of 25 (16%) controls (p = ns). Colorectal varices were seen in 13/41 (31.7%) patients with PHT and none of the controls (p = 0.005). Portal colopathy was present in 15/41 (36.6%) patients with PHT and none of the controls (p = 0.0005). None of the parameters (e.g. aetiology of PHT, Child's class, oesophageal variceal eradication by EST with or without EVL, history of variceal bleeding, grade of oesophageal varices, presence of portal hypertensive gastropathy or gastric varices) predicted the occurrence of colorectal varices and portal hypertensive colopathy. Detection of colorectal varices but not portal hypertensive colopathy was associated with occurrence of hematochezia. PMID- 11398242 TI - Biliary stricture due to tuberculosis of the common bile duct. PMID- 11398243 TI - AIDS cholangiopathy as initial presentation of HIV infection. AB - AIDS related cholangiopathy is a known clinical entity, but most cases described in literature are known cases of HIV infection who have presented with pain abdomen. AIDS cholangiopathy as the initial presentation of HIV infection is very rare. Here we report such a case. PMID- 11398244 TI - Low cost invasive blood pressure monitoring--a new technique. PMID- 11398245 TI - Duplications of the alimentary tract in children. AB - This study was conducted to assess the clinical, diagnostic and therapeutic approach in patients with alimentary tract duplication and review the relevant literature. A retrospective analysis of 28 patients of alimentary tract duplications treated between January 1990-December 1999 was carried out. There was a male preponderance (25:3); 71% of the patients were under 2 years of age. The presenting features were related to the anatomic location. Three patients were diagnosed antenatally. Real time ultrasonography and CT scan was undertaken for preoperative evaluation in the majority of patients. In selected cases a 99m Tc-pertechnetate scan and barium meal follow through examination were also performed. Majority of duplications were in the jejunum and ileum. Twenty percent of our patients had thoracoabdominal duplication as compared to 2% reported in literature. Four patients required to be operated upon in the emergency for related complications. In the remaining patients complete excision of the cyst was done in 21 and only mucosectomy in 3 patients. Because these lesions are rare and can present with a wide range of clinical manifestations or may even be encountered intraoperatively, the appropriate surgical management requires the surgeon to be familiar with the anatomy and clinical characteristics of these lesions. PMID- 11398246 TI - Colorectal cancer in children: case report and review of literature. AB - Compared to adults, colorectal carcinoma is rare in children and has poorer prognosis, due to delay in diagnosis, advanced stage at presentation and aggressive natural history. We describe a case of mucinous adenocarcinoma of the rectum in a eleven year old boy and review the literature regarding colorectal cancer in children. A high index of suspicion and an early diagnosis may lead to an improvement in the dismal prognosis of this disease in children. PMID- 11398247 TI - Surgical management of gallbladder cancer. PMID- 11398249 TI - Diagnosis and management of duodenal obstruction due to renal cell carcinoma. AB - Two cases of duodenal obstruction secondary to renal cell carcinoma are described. One case had delayed metastasis to duodenum four years after right radical nephrectomy and the second case had a large right renal cell carcinoma with duodenal involvement. The possibility of duodenal involvement or metastasis should be kept in mind in any patient presenting with upper gastrointestinal obstructive symptoms and with right sided renal tumour or radical nephrectomy in the past. Whenever suspected, radiography and if required endoscopic assessment should be supplemented to diagnose this condition. Extensive local disease or presence of concurrent metastasis usually rules out the possibility of cure. We describe the clinico-radiological features of this condition along with a review of the literature. PMID- 11398248 TI - Redistribution of gastric Helicobacter pylori after surgery for duodenal ulcer. AB - OBJECTIVES: To study the distribution of H. pylori in the stomach before and after truncal vagotomy and drainage (TV + D) for chronic duodenal ulcer. METHODS: Twenty two consecutive patients studied prospectively. H. pylori positivity detected by urease test and histology (Giemsa stain). One or both tests positive indicated positive H. pylori status. Biopsies taken from antrum, body, fundus and stoma (postoperatively) to document H. pylori status before and 6 months after TV + D. RESULTS: Antral prevalence of H. pylori significantly decreased after surgery while fundal colonization significantly increased. No change was seen in body of stomach. CONCLUSION: There is significant redistribution of H. pylori after TV + D. This has important diagnostic implications in evaluation of postoperative symptoms. PMID- 11398250 TI - Isolated right hepatic duct injury following blunt abdominal trauma: case report. PMID- 11398251 TI - Jaundice and portal hypertension--double trouble. PMID- 11398252 TI - A case of appendiceal-sigmoid fistula of non-diverticular origin. PMID- 11398253 TI - Gall bladder cancer: a disease of Indo-Gangetic belt: the author replies. PMID- 11398255 TI - Factors involved in producing a sustained experimental dermatophyte infection (Trichophyton mentagrophytes). AB - Dr. Jones applied different concentrations of living dermatophyte spores to the surface of uninfected skin under occluded conditions. He then followed the subjects to observe any developing infection and determine the minimum number of spores needed to induce infection. PMID- 11398254 TI - Quality of life after total oesophagectomy for oesophageal cancer. PMID- 11398257 TI - Tinea pedis isolates. AB - An attempt was made to test environmental factors that may influence acquisition of athlete's foot from public places. PMID- 11398256 TI - Dermatophytosis in Vietnam. AB - Professor Taplin describes his experiments producing clinical lesions with 2 endemic dermatophytes encountered in Vietnam during the Vietnam War: the human adapted Trichophyton raubitschekii, possibly a regional form of Trichophyton rubrum that was causing chronic tinea corporis in the Vietnamese, and a zoophilic Trichophyton mentagrophytes that caused highly inflammatory ringworm in the previously unexposed American soldiers. (This particular T mentagrophytes was also the fungus studied by Dr. Jones.). PMID- 11398258 TI - Tinea capitis. PMID- 11398259 TI - Arthroconidia as vectors of dermatophytosis. AB - The characteristic macro- and microconidia produced by dermatophytes in cultures and those living as saprobes are not produced in any dermatophyte lesions observed to date. However, arthroconidia, defined as a genetically programmed disarticulation of septate hyphae to produce a chain of conidia, are produced by dermatophytes, both in culture and in lesions, under certain predisposing conditions. Arthroconidia are seen, for example, within nail-bed infections caused by Trichophyton rubrum; within nail-plate infections caused by Trichophyton mentagrophytes interdigitale; and in the infections of scalp hair by soil or geophilic dermatophytes, where chains of arthroconidia form near or on the surface of the infected hairs. Dr. Rashid discusses his electron microscope studies of the adherence and germination of arthroconidia and the penetration of the germ tubes and initial hyphae into the stratum corneum. PMID- 11398260 TI - Producing experimental lesions of cutaneous candidiasis. AB - Dermatophyte parasitism in normal patients, and even in most immunodeficient patients, is strictly limited to the stratum corneum. The dermatophyte fungi do not readily invade deeper body tissues. Cutaneous candidiasis, however, seems to be strictly an opportunistic infection of ecologically suitable skin by a yeast that is normally present in the gastrointestinal tract and mucosal surfaces of many healthy persons. PMID- 11398261 TI - A review of cellular and molecular events of contact allergic dermatitis. AB - This biochemical review is presented for the first time in a dermatophytosis conference. The intent is to challenge biochemists to explain the formation of these lesions to clinicians. PMID- 11398262 TI - Juvenile tinea capitis. PMID- 11398263 TI - Tinea pedis interdigitalis (interspace). PMID- 11398264 TI - Probabilities of self-diagnosing fungus on the skin--tinea corporis. PMID- 11398265 TI - Dermatophytosis in Vietnam, Part 2. PMID- 11398266 TI - Dermatophytosis in the military. AB - Dr. Elston summarizes the US military experience with dermatophytosis in World War II and Vietnam and then brings us to the present and the current military situation with its increased role in humanitarian assistance. PMID- 11398268 TI - Diaper rash syndrome or dermatitis. PMID- 11398267 TI - Families infected with Trichophyton rubrum. PMID- 11398269 TI - Pharmacokinetics of topical antifungal formulations. AB - The triad of infection--the dynamic equilibrium between pathogen, host, and drug- offers both opportunities and challenges for effectively delivering the right amount of drug to the right place for the right amount of time. The delivery system and drug must cooperate to achieve delivery and substantivity at the target site. Given the amount of variability introduced by the dosage form, it takes plenty of trial and error to get the right drug in the right vehicle so it can reach the targeted tissue. The deeper the target tissue, the greater the complexity of drug delivery and effect. PMID- 11398270 TI - Substantivity of topically applied antifungals on dermatophytic toe webs. PMID- 11398271 TI - Light microscopic study of the effects of terbinafine cream on microconidia: a brief synopsis of modified Knight's Scotch tape skin stripping method. PMID- 11398272 TI - What happens in cidal and static action? AB - Using terbinafine and clotrimazole as examples, Dr. Rashid's electron microscopy images illustrate the differences between fungicidal and fungistatic drugs, respectively. PMID- 11398273 TI - Therapeutic failure and possible involvement of drug resistance: known causes of systemic antimycotic treatment failure. PMID- 11398274 TI - Introducing the syndromes of human dermatophytosis. PMID- 11398275 TI - Bridging the gaps in chronic illness care for children. PMID- 11398276 TI - Simple screen proves highly accurate in identifying children with special needs. AB - Research shows that the relatively small group of children with special health care needs accounts for more than 80% of all child-related health care costs, and there is plenty of evidence that the care of these children is a far cry from optimal. See what steps are being taken to give these children the attention they need, and check out a simple new tool designed to identify children with special health care needs. PMID- 11398277 TI - Predictive modeling care management program produces dramatic bottom line results. AB - It's a new-generation effort, but it has already proven its worth in a large population, producing savings of $1 million to $2 million per month. The effort begins with the development of a constantly evolving registry of high-risk patients. Specialized case management efforts are then focused on stabilizing individuals within this group. See how traditional interventions have been combined with new technology to deliver results. PMID- 11398278 TI - Template for improved management of chronic disease pays off in asthma effort. AB - Change is never easy, but the road map developed by Seattle-based Improving Chronic Illness Care (ICIC) has helped numerous organizations boost their DM outcomes. One of latest organizations to benefit from ICIC's help is Methodist Medical Center in Oakridge, TN. See how this organization has applied ICIC's Chronic Care Model to its burgeoning asthma population. PMID- 11398279 TI - Current status of regulation of ethics in health care in the English-speaking Caribbean. AB - Clinical ethics committees are poorly developed in the English-speaking Caribbean. There is no lack of recognition that there is a problem in the conduct of some health professionals; however, there is a lack of administrative will to deal with the problems. Why there is such a lack of will can be speculated upon and may involve the feeling of a loss of control, and in small communities, the fact that Caesar may be asked to stand in judgement on Caesar. However, the public is increasingly impatient with the stalemate of misconduct and the lack of regulation and is seeking legal remedies or indulging in vilification of the profession to obtain some satisfaction of their grievances. For their own professional health, professionals should demand from administrators that ethical expertise and guidance be made available or they should bypass the administrators and seek to find the necessary expertise and guidance to ensure that this area is no longer ignored and does not remain a convenient and readily available weapon against them. PMID- 11398280 TI - Immunization still saves lives. PMID- 11398281 TI - Salt-induced hypertension in rats alters the response of isolated aortic rings to cromakalim. AB - The effect of cromakalim, an opener of ATP-sensitive potassium (KATP) channel, on precontracted aortic rings from control and salt-loaded rats was studied in Sprague-Dawley rats. Salt-loading experiments involved the induction of hypertension by 6-week feeding of 80 g sodium chloride (NaCl) per kilogram (kg) diet while the control diet had 3 g NaCl per kg diet. Blood pressure and heart rate were determined by cannulation of a femoral artery under urethane/alpha chloralose anaesthesia. Isolated aortic rings were mounted in tissue baths for isometric tension measurement. The sodium-potassium adenosine triphosphatase (Na K ATPase) pump activity was measured by potassium (K+)-induced relaxation (with or without ouabain) following precontraction with 10(-7) M noradrenaline. The KATP channel was studied by measuring the relaxation response to cromakalim, precontracted with either 10(-7) M noradrenaline or 60 mM potassium chloride (KCl). The Na-K ATPase pump appeared to be inhibited during salt loading. ATPase inactivation was found to be ouabain sensitive but did not seem to affect subsequent K(+)-induced contraction. Cromakalim produced relaxation of noradrenaline-precontracted rings from the control rats; rings from salt-loaded rats showed significantly less relaxation than control (p < 0.05) under similar conditions. During K(+)-induced precontraction, cromakalim produced a weak biphasic response in the control rings--an initial relaxation and then a reversal. Cromakalim produced further contraction of K(+)-induced precontraction in the salt-loaded group. The results suggest that ATP-sensitive potassium channels and Na-K ATPase pumps on the vascular smooth muscle membrane may be deactivated in the development of hypertension during salt loading. PMID- 11398282 TI - Stenting of partial and total coronary occlusions in Trinidad and Tobago. AB - In order to evaluate the efficacy and safety of coronary stenting, we reviewed the first 32 consecutive patients (34 vessels) who underwent elective coronary stenting during the period August 1999 to August 2000 inclusive at the Digital Lab installed at the Eric Williams Medical Sciences Complex, Trinidad and Tobago. Aspirin, heparin and ticlopidine were used routinely. Abciximab was used in selected cases (38%). The mean age of patients was 55 +/- 10 years. Eighty-one per cent were male, 52% were hypertensive and 21% were diabetic. Sixty-five per cent had severe angina. Prior Coronary Artery Bypass Grafting (CABG) was performed in 3% and previous Percutaneous Transluminal Coronary Angioplasty (PTCA) in 3%. Multivessel disease was present in 43%. The mean left ventricular ejection fraction was 53 +/- 12%. The culprit lesion was located in either the native left anterior descending (LAD) coronary artery (53%), right coronary artery (RCA) (31%), circumflex artery 13% and saphenous vein graft (3%). The mean baseline diameter stenosis was 91 +/- 9% and this was reduced to 13 +/- 33% after stenting. Procedural success was 100% for 26 partially occluded vs 50% for 8 totally occluded vessels. For the total occlusions, procedural success was inversely related to the duration of the occlusion. There were no cases of death, acute vessel closure, Q-wave myocardial infarction, repeat PTCA or emergent Coronary Artery Bypass Graft (CABG) during and following the procedure. Distal embolization occurred in one patient. The mean duration of hospital stay was one day (for 30 outpatient cases). One patient had recurrence of symptoms with a negative stress test. No patient underwent repeat angiography during the first year of follow-up. Coronary stents were successfully implanted at a tertiary care facility in the Caribbean with low in-hospital morbidity and mortality. Stents markedly reduced the diameter stenosis of the coronary lesion during PTCA. The incidence of clinical restenosis was low. Coronary revascularisation can be successfully achieved by coronary stenting in the Caribbean. PMID- 11398283 TI - Intracoronary stent implantation in Jamaica. The initial experience. AB - Intracoronary stent implantation resulted in the complete or near complete dilatation of high grade occlusions of the left anterior descending coronary arteries in the four patients in whom it was undertaken. Intracoronary stent implantation is a useful adjunct to Percutaneous Transluminal Angioplasty (PTCA) and is applicable in selected patients with symptomatic ischaemic heart disease in a developing country with limited health resources like Jamaica. This is so since financial data presented here document the significant savings this technique (when appropriately utilised) could realise compared to the use of balloon angioplasty alone. PMID- 11398284 TI - Postoperative nausea and vomiting in the recovery room. A report from Guyana. AB - Postoperative nausea and vomiting (PONV) in patients in the recovery room can delay transfer of such patients to the wards, preventing a smooth inflow of patients into the recovery area. The high incidence of these complications in developed countries has led to the introduction of management strategies that may be too expensive for developing countries. An investigation of the incidence and factors associated with PONV in the recovery room of the Public Hospital, Georgetown, Guyana, was undertaken with a view to developing an approach to its management based on local data. Seven hundred and sixty patients were studied. The majority (97%) had general anaesthesia. Simple anaesthetic techniques involving agents of relatively low cost were used. Anti-emetics were infrequently administered. Only twenty-two patients (2.9%) experienced PONV in the recovery room, all after general anaesthesia. Twenty-one of these (95%) had risk factors for PONV in their history. PONV in the recovery room was associated with female gender, gynaecological surgery, extra-abdominal surgery and lack of anti-emetic medication (p < 0.05), as well as prolonged duration of recovery room stay (p < 0.01). A management strategy for PONV in the recovery room involving continued use of present anaesthetic agents, increased use of inexpensive anti-emetic drugs currently available in Guyana, and targeting of patients found to be more at risk is suggested. Increased regional techniques may also be appropriate. PMID- 11398285 TI - Evaluation of neonatal sepsis screening in a tropical area. Part II: Evaluation of intrapartum chemoprophylaxis protocol. AB - The authors report on an analysis of a chemoprophylaxis protocol at the University Hospital of Guadeloupe in the Caribbean. This study comprised 6,060 consecutive deliveries and was initiated to assess the application of an intrapartum chemoprophylaxis protocol, evaluate its results, and try to identify possible necessary modifications to the existing protocol. Although more than 90% of women had at least one bacterial screening (vaginal or urinary) during the last trimester of pregnancy, approximately 75% of mothers who were heavily colonized group B streptococcus (GBS) at delivery were not detected by this systematic screening. As is also reported in other tropical areas where a great proportion of neonatal sepsis occurs in term babies, low birthweight was not a specific risk factor in this study when controlling for other major risk factors such as fever and premature rupture of membranes. Intrapartum chemoprophylaxis was associated with an approximate threefold decrease in the risk of GBS neonatal bacteraemia among at risk deliveries. The results suggest that, in our tropical context, prolonged rupture of membranes of at least 12 hours' duration should be considered as a cause for intrapartum chemoprophylaxis as it accounted for the majority of cases of neonatal bacteraemia that escaped the existing protocol. PMID- 11398286 TI - An audit of perinatal mortality. AB - Perinatal mortality rates are considered in the western world to be a quantitative barometer of maternity care. This 6-year prospective perinatal audit was conducted at a tertiary hospital in order to determine foetal outcome, and the common causes of foetal and early neonatal deaths. Of a total of 30,987 births, there were 469 stillbirths and 391 early neonatal deaths, giving a perinatal mortality rate of 27.7 per 1000 total births. The leading causes of stillbirths were the hypertensive disorders of pregnancy, abruptio placentae, diabetes mellitus, intrapartum foetal distress and lethal congenital anomalies. Neonatal deaths were mainly due to the respiratory distress syndrome (57.8%), birth asphyxia (22.2%) and sepsis (13.5%). A dedicated medical team, including a neonatologist, to manage pre-eclampsia, and more senior obstetric involvement in the labour ward are recommended. PMID- 11398287 TI - A retrospective review of pregnancy outcome after misoprostol (prostaglandin E1) induction of labour. AB - This retrospective study looked at the outcome of using 50-100 micrograms misoprostol once daily to induce labour compared to the outcome of the overall patient population delivered during the same period (1994-1996). During that period 11,255 patients were delivered and 1037 (9.2%) were induced with misoprostol. Results showed a significantly lower mean Caesarean section rate: 9.3% for the misoprostol group versus 13.3% for the overall population (p = 0.002, Odds Ratio (OR) 0.67, 95% CI 0.53, 0.83). The abruption rates were not significantly different: 0.8% for misoprostol versus 0.4% (p = 0.09, OR 1.86, 95% CI 0.81, 4.09). There was more postpartum haemorrhage in the misoprostol group: 5.6% versus 3.5% (p = 0.0006, OR 1.63, 95% CI 1.22, 2.19); a higher incidence of Apgar scores less than 6 at one minute 10.2% versus 7.9% (p = 0.0093, OR 1.33, CI 1.06, 1.65) but not at five minutes 2.9% versus 2.4% (p = 0.674, OR 1.09, CI 0.73, 1.61) and a higher perinatal mortality rate 55/1000 versus 16.3/1000 (p = 0.00, OR 3.5, 95% CI 2.55, 4.80). The rate remained higher but not significantly so when a correction was made to eliminate the high number of intrauterine deaths induced with misoprostol 18/1000 versus 16.3/1000 (p = 0.69, OR 1.11, 95% CI 0.66, 1.84). There were no cases of uterine rupture in either group. In conclusion, there was a significantly lower Caesarean section rate among patients who had once daily misoprostol induction of labour. Close monitoring of the foetus, in patients with misoprostol induction, is needed to detect foetal distress and prophylaxis against postpartum haemorrhage is still mandatory. PMID- 11398288 TI - Facing antibiotic resistance. PMID- 11398289 TI - Antibiotic susceptibility of Clostridium difficile isolates from adult patients at two Jamaican hospitals. Clinical and epidemiological implications. AB - The susceptibility of 39 toxin producing Clostridium difficile isolates from stools of hospitalized patients was determined, by disc diffusion, to six antibiotics. All but one isolate (toxin A negative) produced toxin A and toxin B. A wide variation in susceptibility to clindamycin, tetracycline and chloramphenicol was noted. Erythromycin and cotrimoxazole showed a clear-cut discrimination in resistance and susceptibility, while all isolates were sensitive to vancomycin. Erythromycin sensitive isolates demonstrated a significant association with diarrhoea (60.9%, 14/23, p < 0.001). These strains were predominantly found at the University Hospital of the West Indies (UHWI, 94.1%, 16/17). Strains resistant to erythromycin and clindamycin together were commonly found at the National Chest Hospital (NCH, 68.2%, 15/22). All erythromycin sensitive strains found at the NCH were from patients transferred to that hospital. These findings suggest that there is a common strain of C difficile (erythromycin resistant) at the NCH different from that found at the UHWI; the resistant pattern seen with isolates from the NCH was typical of toxigenic serogroup C strain and could be typed by the the disc diffusion method. Patients at the NCH who were colonized with either of the two strains of C difficile were likely to get diarrhoea, once there was suppression of the normal microflora by antibiotics and colonic overgrowth with C difficile. PMID- 11398290 TI - A clinico-pathological study of Cushing's syndrome at the University Hospital of the West Indies and a review of the literature. AB - Cushing's syndrome is an uncommon but important disease. Twenty-one confirmed cases of spontaneous Cushing's syndrome were documented at the University Hospital of the West Indies over a 21-year period. They were predominantly young females (F:M ratio of 17:4; mean age 25 years and 3 months). The commonest presenting symptoms were amenorrhoea (41%) and obesity (19%). Common clinical features were cushingnoid features (95%), hypertension (76%) and hirsutism (82%). Twenty-nine per cent had frank hyperglycaemia. Cushing's syndrome was due to Cushing's disease in 10 cases, adrenal adenoma in 3 and adrenal carcinoma in 2 cases. In 4 cases with presumed adrenal hyperplasia, the histology was either unavailable or was not consistent with the diagnosis. Two cases appear now to have had the ectopic ACTH Syndrome. Adrenalectomy was the commonest treatment offered. There were no intra-operative or post-operative deaths but recurrence was common after subtotal adrenalectomy in Cushing's disease. Twenty-seven per cent of the patients developed Nelson's syndrome, which was fatal in 50%. Long term hormone replacement therapy was unnecessary after surgery for adrenal adenomas. Treatment of Cushing's syndrome was well tolerated by the patients. PMID- 11398291 TI - Salivary gland lesions: a Jamaican perspective. AB - A retrospective analysis of the spectrum and relative frequency of salivary gland lesions diagnosed in the Department of Pathology, University of the West Indies, Kingston, Jamaica, between 1965 and 1994, is reported. Four hundred and sixty four salivary gland biopsies were received. Of these 99 (21.3%) were non neoplastic and the remaining 365 (78.7%) were neoplasms: 261 (71.5%) were benign and 104 (28.5%) malignant. Benign mixed tumour (BMT)/pleomorphic adenoma (PA) was the most common neoplasm (63.3%) while mucoepidermoid carcinoma (MEC) was the most common malignant neoplasm (9.6%), followed by adenoid cystic carcinoma (ACC) (7.4%). The increased frequency of MEC over ACC is at variance with other reported series but the preponderance of pleomorphic adenoma is consistent. In the major salivary glands, benign neoplasms predominate at a ratio of 3:1, while a higher proportion of minor salivary gland neoplasms was malignant, ratio 1.2:1 (p = 0.003). These data represent the first attempt to document the spectrum of disease related to oral and maxillofacial pathology in Jamaica. PMID- 11398292 TI - An evaluation of medical school education in musculoskeletal medicine at the University of the West Indies, Barbados. AB - At the University of the West Indies, Cave Hill campus, the Freedman and Bernstein musculoskeletal competency examination was administered to 22 medical students during the last month of their final year. Eighty-two per cent (82%) of the students failed to score above the passing score of 73.1%. Nineteen of the twenty-two students had taken an orthopaedic elective or rotation during their final two years. The questions were also categorized as Anatomy, Trauma and General Orthopaedics. All students failed to score above the passing score in Anatomy. Sixty-four per cent (64%) failed in Trauma and 45% failed in General Orthopaedics. This study suggests that inadequacies in medical school preparation do exist at this campus of the University of the West Indies and the findings mirror those at medical schools in the United States of America. PMID- 11398293 TI - Medical students' evaluation of their pathology clerkship experience. AB - As part of a recent trend among faculties in educational institutions to review and upgrade their courses and teaching methods, the Department of Pathology introduced a questionnaire for assessment of the pathology clerkship experience by the medical students. This was to be completed at the end of their rotation. An evaluation of the results of an entire class indicates that, for the most part, the responses were positive with 82% considering the experience to be of great benefit and 17% of reasonable benefit. The responses were generally more favorable to Anatomical Pathology and Haematology than to Chemical Pathology. The study identified areas where modifications in teaching could be considered, and ways to improve the questionnaire, making it more useful for future assessments. PMID- 11398294 TI - Images and diagnoses. Trichoepithelioma (epithelioma adenoides cysticum). PMID- 11398295 TI - Images and diagnoses. Carcinoma of the pancreas with Sudeck dystrophy of fingers. PMID- 11398296 TI - An atypical aggressive Kaposi's sarcoma in a HIV-negative bisexual man. AB - We describe a case of Kaposi's sarcoma in a previously healthy 35-year-old bisexual West Indian man of African descent who was seronegative for the human immunodeficiency virus (HIV) and human T-cell lymphotropic virus-1 (HTLV-1) and who presented with extensive mucocutaneous lesions, weight loss, visceral and generalized lymph node involvement, poor response to combination therapy with vinblastine and interferon alfa-2a, and a short survival of eight months from the onset of illness. This is the first documented case of Kaposi's sarcoma in Dominica. The presentation is unusual in that it is similar to the aggressive and disseminated Kaposi's sarcoma seen only in AIDS and the florid variant of the endemic disease in young men in Equatorial Africa. PMID- 11398297 TI - Clear cell sarcoma of soft parts. A case report and review of the literature. AB - The case of a 25-year-old woman with recurring and metastasizing clear cell sarcoma of soft parts (CCS) is presented. The clinical setting and pathological appearance were in agreement with the original cases described. This report serves to highlight the clinico-pathological features of this rare and aggressive malignancy with specific reference to prognostic markers and treatment. PMID- 11398298 TI - Helicobacter pylori infection in the Caribbean: update in management. AB - Helicobacter pylori infection of the stomach is one of the commonest chronic infections worldwide and in the Caribbean, over 50% of the population are affected. H pylori is probably transmitted from person to person by oro-faecal and oro-oral means. H pylori is directly associated with peptic ulcer disease, chronic antral gastritis, gastric carcinoma and B-cell lymphoma of the stomach. In patients with peptic ulcers and H pylori infection, eradication of infection with antibiotics significantly decreases recurrence of ulcers. All patients with H pylori related disease should be tested and treated if positive. The treatment of H pylori infection has evolved over the years but at present triple therapy which includes two antibiotics is recommended. PMID- 11398299 TI - The MMR vaccination and autism: a lay person's contribution. PMID- 11398300 TI - Ketoprofen, acetaminophen, and placebo in the treatment of tension headache. AB - Seven hundred three subjects completed a randomized, double-blind, parallel group, single-center study comparing the single-dose efficacy of ketoprofen 12.5 mg, ketoprofen 25 mg, acetaminophen 1000 mg, and placebo in the treatment of tension headache. For the primary efficacy variable, 4-hour sum of pain relief intensity differences, ketoprofen 25 mg was significantly superior to placebo. Ketoprofen 25 mg also demonstrated superior pain relief in the first hour after dosing, and the time to meaningful pain relief was significantly shorter for the ketoprofen 25-mg group. Ketoprofen 12.5 mg proved to be significantly superior to placebo for pain relief intensity difference and pain relief at 3 hours, global assessment of medication at 4 hours, and for time to onset of meaningful pain relief. The data suggest a dose response for ketoprofen 12.5 mg and 25 mg. Acetaminophen 1000 mg proved to be numerically more favorable than placebo in most variables, but could not be separated from placebo with statistical significance. In spite of possible inflation of the placebo response due to sensitivity limits of the study, ketoprofen 25 mg demonstrated a more rapid onset of analgesia compared to acetaminophen 1000 mg, and patients' global assessment rated ketoprofen 25 mg higher than acetaminophen 1000 mg. PMID- 11398301 TI - The surgical management of chronic cluster headache. AB - OBJECTIVE: Chronic cluster headache occurs in less than 10% of cluster headache sufferers, but remains an intractable medical problem. Surgical treatments have also been limited in their effectiveness. The authors describe their experience with attempted surgical amelioration of chronic cluster headache. DESIGN: Twenty eight patients, including two with bilateral cluster headache, underwent 39 operations for microvascular decompression of the trigeminal nerve, alone or in combination with section and/or microvascular decompression of the nervus intermedius. Follow-up averaged 5.3 years. RESULTS: Initial postoperative success described as 50% relief or greater was achieved in 22 (73.3%) of 30 first-time procedures and greater than 90% relief in half (15 of 30) of these. Long-term follow-up saw this success rate (excellent or good) drop to 46.6%. Repeat procedures have little success, with 7 of 8 failing at long-term follow-up. Morbidity and neurological deficit from the operations was minimal. CONCLUSIONS: Chronic cluster headache remains a debilitating and poorly controlled syndrome. Although various surgical treatments have had limited success, microvascular decompression of the trigeminal nerve with section of the nervus intermedius compares very favorably to other destructive techniques without the accompanying neurologic deficits. It is, therefore, our recommendation as the first-line operative treatment of chronic cluster headache. PMID- 11398302 TI - Migraine prevalence: a literature review. AB - Worldwide epidemiological studies provide estimates of migraine prevalence. The International Headache Society (IHS) diagnostic criteria have permitted more reliable and generalizable study estimates. In this review, recent population based reports of IHS-defined migraine are discussed to update the current epidemiological literature, to highlight methodological challenges, and to encourage greater public awareness of this frequently undiagnosed disabling disorder. PMID- 11398303 TI - Migraine therapy: development and testing of a patient preference questionnaire. AB - OBJECTIVES: To develop and test a patient preference questionnaire in migraine. METHODS: A user-friendly, self-administered questionnaire was developed to assess the relative importance of aspects of migraine therapy to patients. It was tested in a convenience sample of 66 migraineurs. The questionnaire has five sections: patient preference, migraine history, demographics, usual behavior during an attack, and migraine impact on cognition and functionality. It employs a special reverse-ranking technique to quantify preferences. RESULTS: Sixty-six migraineurs participated: 86% were women, 53% had not consulted their doctor about migraine in the previous year; and only 51% took prescribed medication, the majority (52%) at the start of a migraine. "A medication that can be taken any time during a migraine attack" was ranked most important by 20% of the migraineurs, one that decreases pain from the unbearable level in 30 minutes by 17%, and one that relieves pain completely within 2 hours by 15%. Not being able to think or concentrate with severe symptoms was reported by 71%. Optional activities were impeded by migraine in 83% of subjects, but required activities in only 60%. CONCLUSIONS: The patient preference questionnaire is readily completed by subjects and reveals that the dimensions of relief preferred by patients can be addressed by a self-report questionnaire. PMID- 11398304 TI - Cerebral blood flow and CO2 reactivity in interictal migraineurs: a transcranial Doppler study. AB - There is still some controversy about alterations in velocity of blood flow and in cerebral vasomotor reactivity of intracranial arteries in migraineurs during the interictal phase. By means of simultaneous bilateral transcranial Doppler ultrasonography we, therefore, assessed intracranial blood flow velocities and cerebrovascular reactivity to carbon dioxide of all three basal brain arteries in 20 migraineurs during the interictal phase and 30 nonheadache-prone control subjects. Mean blood flow velocities were higher in migraineurs than in controls in all three arteries on both sides, with a significant difference (P < 0.05) for the right anterior cerebral artery and middle cerebral artery under basal conditions and for the right posterior cerebral artery during hypercapnia. Similarly, the cerebrovascular reactivity to carbon dioxide was always higher in patients than in controls, with a significant difference for the left anterior and the right middle cerebral arteries (P < 0.05) and the right posterior cerebral artery (P < 0.01). The broad overlap of cerebrovascular blood flow velocities and CO2 reactivities in patients and controls precluded identification of values diagnostic of migraine. Nevertheless, transcranial Doppler ultrasonography offers the opportunity to noninvasively monitor cerebral blood flow parameters and, therefore, represents a valuable tool for vascular research in migraine. PMID- 11398305 TI - Dihydroergotamine nasal spray in the treatment of acute migraine. AB - OBJECTIVE: To evaluate the efficacy and safety of dihydroergotamine (DHE) nasal spray in patients suffering from common or classical migraine. METHODS: In a double-blind parallel-group study, 52 outpatients with migraine were randomly allocated to DHE nasal spray or to placebo. Two puffs, one in each nostril, was taken as an initial dose (resulting in either 0.5 or 1 mg of DHE), followed by another puff (0.5 mg) after 30 and 60 minutes, if necessary, achieving a maximum dose of 2 mg for patients of the DHE 1-mg group or of 1.5 mg for patients of the 0.5-mg group. Four consecutive attacks were thus treated. The efficacy analysis was done for observed cases. The main outcome measure was reduction of the severity of the attacks. RESULTS: No differences were observed in the migraine characteristics or the number of treatments of the patients from the different groups. Dihydroergotamine 1 mg tended to provide better relief than 0.5 mg, although the effect was not statistically significant. Patients taking DHE used less rescue medications, with a dose-dependent effect. Side effects were reported by four patients receiving DHE but not placebo. The tolerability of the drug was assessed as good by 94% of the patients. CONCLUSION: These findings suggest that DHE nasal spray is well tolerated and has dose-dependent efficacy in migraine. PMID- 11398306 TI - Prostaglandin analog mechanisms are not effective in refractory chronic cluster headache. AB - Prostaglandin E analogs have been shown to be effective in the treatment of refractory trigeminal neuralgia in patients with multiple sclerosis. Prostaglandin E inhibits the functions of T lymphocytes which are involved in the pathophysiology of cluster headache. Therefore, a double-blind, placebo controlled, crossover study on the efficacy of misoprostol in chronic refractory cluster headache was performed. Eight patients were treated with 600 micrograms misoprostol and with placebo for a 2-week period. No differences in attack frequency, intensity, global impression, and side effects could be detected, suggesting that prostaglandin E analogs are not effective in the treatment of chronic cluster headache. PMID- 11398307 TI - Headaches and papilledema secondary to dural arteriovenous malformation. AB - A 21-year-old developed progressive headaches and pulsatile tinnitus. He was found to have papilledema and a pulsatile bruit. A dural arteriovenous malformation was not recognized on brain CT or MRI, but was well documented on magnetic resonance angiography and cerebral angiography. Dural malformations draining into the superior sagittal sinus can cause increased intracranial venous pressure and secondarily increase intracranial pressure even without a mass effect. PMID- 11398308 TI - Amlodipine for migraine prophylaxis. AB - Migraine is among the most common neurologic disorders encountered in clinical practice. In the general US population, the annual incidence has been calculated to be approximately 250 per 100,000 with a point prevalence of 10%. Females are affected more than males. A variety of prophylactic and abortive medications are being used for treatment and several are being studied in clinical trials. Calcium-channel blockers are frequently used prophylactic medications. We report two patients with migraine successfully treated with amlodipine (Norvasc, Pfizer, Inc), a slow calcium-channel blocker. To our knowledge, these are the first reported cases of amlodipine used in migraine prophylaxis. PMID- 11398309 TI - Migraine and coeliac disease. AB - The pathophysiology of migraine remains unclear. To incriminate a comorbid factor is always hypothetical, even if suppression of this factor appears to prevent the migrainous attacks. In our patient, treatment of coeliac disease coincided with total disappearance of severe migraine attacks. Moreover, the coeliac disease was first revealed during the evaluation of a migraine with aura. PMID- 11398310 TI - Cluster headache dependent upon octreotide injection. PMID- 11398311 TI - Efficacy of sumatriptan in chronic paroxysmal hemicrania. PMID- 11398312 TI - [Nutritional consideration for changes in dietary habit and health promotion practices in community health care; from the view point of selenium]. AB - The Japanese recommended dietary allowances (RDA) for major and some minor nutrients were revised in 1999, and included those for trace elements such as selenium. The requirement of selenium in animals was first recognized in 1957. It has been shown that cellular glutathione peroxidase (GPx) contains selenium but it was subsequently revealed that selenium has diverse biochemical effects, rather than simply functioning in the enzyme. At least twelve different selenoproteins have been identified. The role of selenium has been known as antioxidant, and non-antioxidant mediated through these enzymes. Now, selenium is well recognized as a preventive factor for cancer and cardiovascular diseases. Several dietary studies have shown that the selenium intake in Japan is adequate. One study estimated daily selenium intake to be 104.2 micrograms/day for adults. This value was 2 or 3 times higher than the lower limit of the safe range of dietary selenium (40 micrograms/day for men and 30 micrograms/day for women) estimated by WHO, and also exceeded the newly established RDA of 55-60 micrograms/day for men and 45 micrograms/day for women by the Japanese Public Health Council. However, the established RDA for selenium is tentative because of a lack of information on the 1) chemical forms of selenium in food, 2) differences in absorption rate and bio-availability in the chemical forms, and 3) interactions with other metals and trace elements. There are two potential problems concerning selenium nutrition in Japan. The first problem is that rice, which is the Japanese staple food, contains less than 0.05 microgram/g selenium whereas U.S. rice contains more than 0.3 microgram/g, probably due to differences in soil chemistry. The second problem is that although studies have shown that seafood, fish, shellfish and oysters, contain high levels of selenium (0.4-0.5 microgram/g), these being the main selenium source for Japanese, the bio availability in fish is low. Thus, it is likely that the selenium status of those Japanese who eat an imbalanced diet is not sufficient or is not optimal even if the intake exceeds the RDA. Further studies are needed so that community health care specialists have available appropriate knowledge on the role of trace nutrients, including selenium, in human nutrition and health, to promote proper nutritional practices in the community. PMID- 11398313 TI - [Development and evaluation of a QOL questionnaire for elderly subject living in a community]. AB - A comprehensive, basic and simple QOL questionnaire for elderly subjects living in a community was developed, and its validity and reliability were examined. The subjects were 2944 individuals of 65 years or older living in 5 areas of metropolitan Tokyo and in a town of Aichi Prefecture. The QOL questionnaire with 19 questions was developed based on the component of QOL by Lawton and concept of QOL by Koyano. The questionnaire consisted of 6 subscales (daily activity, satisfaction with health, satisfaction with human support, satisfaction with economic state, symptom of depression and positive mental attitude). Factor analysis revealed that the 19 questions could be clearly separated into 6 components in Tokyo and Aichi districts with total variances of 70.8% and 78.4%, respectively. Scores of daily activity and positive mental attitude were significantly lower with older subjects in both men and women. However, scores for other subscales did not differ with age. Primary factors which are considered to affect QOL were compared with the 6 QOL subscales of this study. Being an outpatient had a significant relation to daily activity and satisfaction with health, presence of a spouse to satisfaction with human support, depressive state, positive mental attitude and possession of ones own room to satisfaction with economical state, and belief in religion to positive mental attitude. The results suggest that the present questionnaire include the basic components necessary for evaluation of QOL in elderly subjects living in a community. Further research is required to examine the validity of this questionnaire with correction of questions. PMID- 11398314 TI - [Relations between taking contraceptive pills, condom use and sexually transmitted disease history among female sex workers]. AB - PURPOSE: The aim of this study was to analyze how taking contraceptive pills and condom use are related to each other, and their links to sexually transmitted disease (STD) history among female sex workers. METHOD: A cross-sectional study was performed by means of a standardized questionnaire self-administered by sex workers who attended a STD Clinic in Osaka between 1st April, 1998 and 31st March, 1999. They came to the clinic for routine screening for STDs and HIV (Human Immunodeficiency Virus) infection, and for treatment. The responses of a total of 92 sex workers, who are basically expected to use condoms and identified themselves as "sex workers at bath houses", "sex workers in former red light districts", "sex massage providers on call" and "call girls" on the questionnaires, were analyzed. RESULT: The proportion of sex workers who were diagnosed with chlamydial infection either currently or over the previous year was higher among sex workers who answered they take contraceptive pills "regularly" (pill users) than in those who answered "irregularly" or "not" (non pill users). This finding was statistically significant for those whose education was less than 12 years, whose professional career as a sex worker was 1 year or more, and whose average daily income was 50,000 yen (US$450) or more. Except for the group whose income was less than 50,000 yen, the proportion of sex workers in all groups who were diagnosed with chlamydial infection was greater, although the difference was not statistically significant, among pill users than among non users. The proportion of sex workers who use condoms "regularly and correctly" was significantly higher for non-pill users than for those using the pills. Multiple logistic regression analysis showed that pill use was significantly related to condom "not or irregular or incorrect" use (Odds ratio: 4.43, CI 1.12 17.60). The proportion of sex workers with a chlamydial infection history was significantly lower with those using condoms "regularly and correctly" than in those who do not use condoms or employ them "irregularly or incorrectly" (Odds ratio: 0.08, CI 0.01-0.68). CONCLUSION: This study showed the proportion of sex workers diagnosed with chlamydial infection to be higher among pill users than among non-users and the proportion of sex workers who use condoms "regularly and correctly" was higher among non-pill users than among users. These findings suggest that irregular, incorrect or non use of condoms among pill users may cause chlamydial infection. PMID- 11398315 TI - [Coronary risk appraisal for primary prevention of coronary heart disease in a community]. AB - PURPOSE: In discussion on application of "National Health Promotion Toward 21st Century in Japan" in Kanagawa prefecture, it was noted that the age-adjusted mortality rate of death from ischemic heart disease in this part of Japan was higher than that for the whole nation in 1996. To facilitate development of a strategy for primary prevention of coronary heart disease (CHD), the present study was conducted to predict 2-yr incidence of CHD and decrease with simulations assuming improvement in CHD risk factors. METHODS: Using CHD risk prediction algorithm; the Weibull accelerated failure regression model based on the Framingham Heart Study, a 2-yr incidence of CHD was predicted for 1652 residents (515 male, 1137 female) on the basis of results of a health check up in 1998. We then estimated the probable decrease in CHD recalculated assuming decrease in total cholesterol (TC), increase in HDL-cholesterol (HDL-C), decrease in systolic blood pressure (SBP), or quitting the smoking habit. RESULTS: 1. The 2-yr probability of developing CHD for men free of heart disease was 2.79 +/- 2.17%, and that for men who had heart disease was 10.25 +/- 2.17%. The 2-yr probability for women free of heart disease was 16.80 +/- 14.40%, and that for women who had heart disease was 3.66 +/- 1.09%. As the reported probability of developing CHD in the U.S.A. is remarkably higher than in Japan, the fact that the present model was based on American data explains why these predicted probabilities are higher than values reported from Japanese cohort studies. 2. For men free of heart disease, a strategy for high risk case such as a decrease in TC and an increase in HDL-C, or quitting the smoking habit, was more effective than a population-based strategy. For women free of heart disease, the population based strategy was more effective. 3. Women more than 60-yrs old who had a high 2 yr probability of developing CHD were divided into three groups; high, middle, and low risk. The mean body weight, mean body mass index, mean diastolic blood pressure, and mean blood glucose in the high risk group were significantly higher than the values in the other groups. Decrease in systolic blood pressure was a more effective strategy for decrease in CHD incidence in the high risk group than in the other groups. CONCLUSIONS: CHD risk prediction of this type may be considered useful for setting target CHD risk factors and for focusing interventions to prevent CHD effectively. PMID- 11398316 TI - [Decline in birthweight-specific infant mortality rates of singleton livebirths in the 1990s in Japan]. AB - OBJECTIVE: This study investigated the decline in birthweight-specific infant mortality rates in Tohoku, Tokai and Kyushu regions between 1989 to 1990 and 1995 to 1998. METHODS: Information for both births and infant deaths from any diseases were collected for two periods in the Tohoku, Tokai and Kyushu regions. The first involved a cohort of singleton livebirths (n = 404, 158) in the three regions in 1989, whose birth certificates were linked with infant death certificates. The second was for singleton livebirths (n = 1,503,230) in the same regions between 1995 and 1998, and infant deaths from any diseases were identified based on vital statistics. RESULTS: Overall infant mortality rate from any diseases declined 16%, from the 3.86/1000 of the first group to the 3.24/1000 of the second group. A shift to a lower birthweight distribution weakened the improvement in the overall mortality rate, so that birthweight standardized mortality rates were reduced more than crude ones. Based on birthweight adjusted relative risk, neonatal, postneonatal and infant mortality rates for each birthweight group were improved by almost uniform ratio, that is 31%, 12% and 23% respectively. Improvement of survival of infants with birthweight of a 2500 or more accounted for most of the decline in the infant mortality rates in the 1990s. Mortality rates from "certain conditions originating in the perinatal period" and "congenital anomalies" decreased remarkably, whereas the sudden infant death syndrome increased. CONCLUSIONS: We conclude that reduction of mortality rates in the infant period in Japan in the 1990s was attributable to decline in birthweight-specific mortality rates and weakened by a shift to a lower birthweight distribution over time. Normal birthweight infants showed the greatest improvement. PMID- 11398317 TI - [Care networks of visiting nurse agencies before the long-term care insurance act]. AB - PURPOSE: To describe how organizational background impacts on care networks of visiting nurse agencies. METHODS: A questionnaire was sent to all visiting nurse agencies (3,178) in Japan, and 1,548 responded. Questions were asked about the agency's background, management system, and cooperation with other community care agencies. FINDINGS: Seventy-nine percent of visiting nurse agencies were affiliated to medical organizations consisting of hospitals, clinics, or medical nursing facilities, The exceptions cooperated with various types of community service agencies, including home care agencies, assisted device agencies, housing improvement agencies, or volunteer services. The visiting nurse agencies had more frequent contact with housing improvement agencies when home care aid agencies were included within the same organizations. The results suggest that visiting nurse agencies can be divided into two types with reference to the long-term care insurance system. One has strong cooperative relationships with hospitals, and the other with community care agencies. PMID- 11398318 TI - [Analysis on the relationship between use of health check ups and medical care by elderly patients. A study on urban cities with high health check-up rates]. AB - The aim of this study was to evaluate the influence of health examination provided by municipalities in Japan on the use of medical care by elderly patients aged 70 and above. The study was carried out to compare health indicators of cities with high and low usage rates for health check ups over 14 years. The indicators of health service utilization included mean bed days, mean inpatient fees, and mean outpatient fees in 1983, 1988, 1993 and 1996. The data for these indicators were collected from all 12 Japanese cities, where more than 50% of the target population had annual health check-ups in 1983, 1986, 1989 and 1992, as the subjects of high rate cities. All other 134 cities belonging to the same prefectures with the 12 cities were included as the control low rate cities. The correlation coefficients for all the 146 cities showed that usage rates of health check ups had an inverse relationship with mean bed days and mean inpatient fees for the elderly population in all the fiscal years 1983, 1988, 1993 and 1996. In nine out of the ten prefectures, mean bed days and mean inpatient fees were lower in the high rate cities as compared to the low rate cities for all the fiscal years compared. The bed days in the 12 high rate cities were 72% of those in 134 low rate cities in 1983, and the percentages were 66%, 72%, and 78% in 1988, 1993 and 1996 respectively. No remarkable differences in mean outpatient fees were observed between the high and low rate cities. The differences in use of medical care by inpatients suggest that providing preventive health services would decrease the demand by the elderly and result in a more efficient use of health resources. PMID- 11398319 TI - [Comparisons of factors affecting voiding disorders between patients with benign prostate hyperplasia and volunteers]. AB - PURPOSE: The prostate size and motivation to visit clinics were investigated in patients with prostate hyperplasia. OBJECTS AND METHODS: One hundred ninety-five patients who had urinary symptoms and visited our outpatient clinic between September 1994 and October 1999 and 268 age-matched volunteers in Mitaka City who underwent a medical examination of the prostate in June 1997 were compared. International Prostate Symptom Score (IPSS), Quality Of Life Score (QOL score), residual urine volume, prostate volume and urinary flow rate were measured. RESULTS: The prostate volume of the volunteers was 20-25 cm3 irrespective of the age. The prostate size of the outpatients was larger than that of the volunteers for every age group. IPSS and QOL score were significantly higher in the outpatients than in the volunteers. Diurnal urinary frequency and sense of residual urine contributed to the discrimination index of the two groups more significantly than the other scores. There was a significant correlation between prostate volume and residual urine volume. The score of weak urinary steam was inversely and significantly correlated with peak urinary flow rate. CONCLUSIONS: There was no age-related enlargement of the prostate gland. The prostate gland was significantly larger in the patients than in the volunteers even in those in their fifties. Urinary frequency and sense of residual urine are important factors for men to seek and receive medical care. PMID- 11398320 TI - [Analysis of clinical costs for inpatients with the urologic diseases in 1997]. AB - OBJECTIVES: Because of recent increases in health care costs, reformation care cost system similar to the disease related group/prospective payment system (DRG/PPS) was attracted attention in Japan. We analyzed the costs for inpatients with urologic diseases in our institution and compared them with the costs reported in Japanese trials of DRG/PPS. MATERIALS & METHODS: Subject consisted of 363 consecutive urologic diseases inpatients treated in our hospital in the period between April 1 1997 and March 31 1998. Of these 107 had bladder tumor, 79 BPH (benign prostatic hyperplasia), 34 prostate tumor, 32 renal tumor, 17 in urinary stone (excluding cases with bladder stones) and 94 cases in others. The total hospital charge for each case was calculated, and the averages and standard deviations for each disease were analyzed. We analyzed the relationship between hospital charges for each disease, and complications, the degree of disease, age and length of stay (LOS). Health care costs are assessed in unit, each of which is the equivalent of 10 yen. RESULTS: The average of total hospital charges was 108,841.6 points for bladder tumor, 61,692.2 points for BPH, 96,825.3 points for prostate tumor, 188,459.0 points for renal tumor, and 66,052.2 points for urinary stones. The standard deviation was 99,611.3 points for bladder tumors, 16,712.9 points for BPH, 65,419.3 points for prostate tumor, 204,389.6 points for renal tumor and 30,081.7 points for urinary stone. There was a positive correlation between hospital charge and LOS in most diseases. The average hospital charges for each disease were much higher than the reimbursement charges of the trial Japanese DRG/PPS. DISCUSSION: Shortening of LOS using established clinical pathways reduces health care costs. However, in Japan, the wide range of preliminary results suggests that it may be too early to introduce the DRG/PPS system for health care cost. Furthermore nationwide investigations are needed before the introduction of the DRG/PPS system into Japanese health care reimbursement system. PMID- 11398321 TI - [Sexual function in men with lower urinary tract symptoms]. AB - PURPOSE: Recently there have been several alternatives not only to improve symptoms but to retain an acceptable quality of life as well as to reduce the complications. Therefore the objective of this study was to evaluate quantitatively and qualitatively the degree of erectile dysfunction in the population of men with lower urinary tract symptoms (LUTS). MATERIALS AND METHODS: Total 252 men with LUTS were investigated using the International Prostate Symptom Score (I-PSS) and sexual function inventory (SFI) consisting of sexual drive, erection and ejaculation. Spearman's rank order correlation was used to determine the degree of any correlation between age, the total I-PSS, the individual I-PSS questions and the various sexual function scores. RESULTS: 208 patient data were available for analysis. There were poor function patients in 67.8% for sexual drive, in 46.2% for erection and in 47.1% for ejaculation. On the other hand, 24% considered their sexual drive to be a big or medium problem, 20.7% for erection and 18.3% for ejaculation. Overall, 27.4% of the men were mostly or very dissatisfied with their sex life. There was a significant correlation between a patient's age and his score for each of the three sexual variables (p < 0.05). Furthermore, a significant correlation was noticed between the total I-PSS and the SFI score (p < 0.05). It might be suggested that the more symptomatic a patient for LUTS, the poorer his sexual function will be. CONCLUSIONS: About 20% men with LUTS are bothered by their sexual symptoms. Erectile dysfunction in dependent of age and the extent of LUTS. Consideration of the high population of erectile dysfunction in men with LUTS is necessary to the treatment of their urinary symptoms. PMID- 11398322 TI - [Ectopic ureter in 54 children]. AB - PURPOSE: We reviewed 54 pediatric patients with ectopic ureter treated at our institution from 1975 to 1999. MATERIALS AND METHODS: Our series comprised 40 female and 14 male children, with age from 1 month to 11 years. Clinical records of the patients were reviewed retrospectively. RESULTS: Chief complaint was urinary incontinence in 24, high fever in 18, abdominal mass in 6, scrotal swelling in 3 and growth retardation in 2 patients. Two patients were found to have ectopic ureters without symptom during urological work-ups for their anorectal anomaly. The ectopic ureters opened into vagina in 19, vestibulum in 8, bladder neck in 7, urethra in 17, seminal vesicle in 2 and ejaculatory duct in 1 patient (s). Treatment was ureterocystoneostomy in 30, nephroureterectomy in 19, hemi-nephroureterectomy in 2, and ureteral ligature in 1 patient (s). Postoperatively, most of the patients became symptom free except for 6 patients in whom urinary incontinence was not cured due to mal-development of the bladder neck and sphincter, and due to Gartner's duct cyst. CONCLUSION: Urinary incontinence and urinary tract infection are most frequent presentations of ectopic ureter in children. Although most of the patients are cured with ureterocystoneostomy or nephroureterectomy, some incontinent girls continue to have urinary incontinence due to mal-development of the bladder neck and sphincter or Gartner's duct cyst. PMID- 11398323 TI - [A case of rectal bladder construction by Duhamel procedure for vesical extrophy]. AB - A 4 year old boy, who underwent 2 times of unsuccessful primary closure of the extrophied bladder at 7 days and 5 months after birth, was treated with construction of a rectal bladder with sigmoid-pull through by Duhamel procedure. Vas deference, seminal vesicles and prostate were not found at the cystectomy operation. Epispadias urethral mucosa was removed and phalloplasty was performed using the ventral hooded foreskin. The results were cosmetically and functionally excellent. He could keep continence and his upper urinary tract was normal at 6 months after the operation. PMID- 11398325 TI - [Hydronephrosis caused spontaneous evacuation of many small calculi in a caliceal diverticulum: a case report]. AB - We experienced a curious case that hydronephrosis had caused spontaneous evacuation of calculi in a caliceal diverticulum. A 50-year-old woman was visited to our department complaining of right lower abdominal pain. Several right ureteral stones, right hydronephrosis and a great numbers of small round-shaped calculi in a right caliceal diverticulum were diagnosed. Many of the calculi moved from the diverticulum to ureter, and were evacuated spontaneously. The month after, the right hydronephrosis had already subsided and almost all of the calculi had disappeared. It was supposed that a hydronephrosis had widened the narrow channel between the diverticulum and the calyx and enabled the small calculi to pass through. Recent reports show that effectiveness of ESWL for symptomatic caliceal diverticular calculi is doubtful, because the narrow channel hinders the passage of stone fragments. However, this case suggests that an artificial hydronephrosis created by retrograde occlusion ureteral balloon catheter may lead to good drainage of gravel and better stone-free rate of caliceal diverticular calculi treated by ESWL. PMID- 11398324 TI - [A case of pheochromocytoma with von Recklinghausen's and review of 67 Japanese cases]. AB - A 19-year-old woman with von Recklinghausen's disease was referred to our hospital because of right adrenal pheochromocytoma. The tumor was detected incidentally with the abdominal ultrasonography when she complained epigastralgia to the home doctor who treated her hypertension. Plasma and urinary catecholamines level were elevated. The tumor was removed by laparoscopy assisted adrenalectomy without pneumoperitoneum. The resected specimen was 35 x 60 x 75 mm in size and weighed 70 g. Pathological diagnosis was adrenomedullary pheochromocytoma. Postoperative course was uneventful. She has been well with no signs of recurrence after 7.5 years. We reviewed 67 Japanese patients previously reported as von Recklinghusen's disease with pheochromocytoma. Of the 60 patients whose details were described, 16.7% had metastases and pathological malignancy from pheochromocytoma. PMID- 11398326 TI - [Comparison of in vitro antimicrobial activities of ofloxacin, levofloxacin, ciprofloxacin, and sparfloxacin against various mycobacteria]. AB - In vitro antimicrobial activities of ofloxacin (OFLX), levofloxacin (LVFX), ciprofloxacin (CPFX), and sparfloxacin (SPFX) were compared against various mycobacteria using the agar dilution method with 7H11 medium, and the following results were obtained. (1) These four new quinolones showed excellent antimicrobial activities against M. tuberculosis, M. kansasii, and M. fortuitum. (2) SPFX was most active against slowly growing mycobacteria. The activity against M. tuberculosis was in the order of SPFX > CPFX > LVFX > OFLX. The activity against M. kansasii was in the order of SPFX > LVFX > OFLX > or = CPFX. (3) On the other hand, CPFX was most active against rapidly growing mycobacteria. The activity against M. fortuitum was in the order of CPFX > SPFX > LVFX > OFLX. Considering the in vitro antimicrobial activities and the pharmacokinetics of these four drugs, they could achieve favorable clinical outcomes for all the patients with pulmonary infection due to M. tuberculosis or M. fortuitum and some of the patients with pulmonary infection due to M. kansasii or M. chelonae. PMID- 11398327 TI - [A nutritional investigation of homeless patients with tuberculosis]. AB - A retrospective case-control study was performed with TB patients who were admitted to our hospital over the two years from Jan. 1997 to Dec. 1998 and healthy men who underwent a health screening in April 2000 in the same hospital. Thirty-two non-homeless TB patients (the first control group) and 32 healthy men (the second control group) were matched with 32 homeless TB patients according to age. All 3 groups were male. Total protein, albumin, cholesterol, cholinesterase, hemoglobin level and lymphocyte count on admission were significantly lower in the homeless patients than in the non-homeless patients and healthy men. Albumin, cholesterol, cholinesterase, hemoglobin level, white blood cell count and lymphocyte count on admission were significantly lower in non-homeless patients than healthy men. Height, weight and body mass index were significantly lower in the homeless patients than in the healthy men. However, there were no significant differences in these body characteristics between the homeless and non-homeless patients. Twenty-five percent of homeless patients died during hospitalization, compared with 6.3 percent of non-homeless patients. Lymphocyte counts among homeless patients who died during hospitalization were significantly lower than among those who survived during hospitalization. Total protein, albumin, cholesterol, cholinesterase, hemoglobin level and weight were lower in patients who died than in those who survived, although the differences were statistically not significant. PMID- 11398328 TI - [Analysis of factors related to the high incidence of tuberculosis in the city of Wakayama--analysis according to age groups, and sputum test results]. AB - Over the last 20 years, the decrease in the incidence of Tuberculosis (TB) in Japan has slowed down. As of 1999, the incidence rate was 34.6 per 100,000 population in Japan, which was higher than that of the other developed countries, and the incidence rate in the city of Wakayama, one of the prefectural capital cities in Japan, during the same period was 42.9 per 100,000 population. We investigated the causes of this high incidence rate of TB in Wakayama City according to the analysis by age groups and sputum test results when patients are newly registered. Comparing our data during the period from 1.1.1998 to 12.31.1999 with data during the same period in the whole country and the rest of Wakayama Pref., the following results were obtained. Observing by age-groups, the incidence of TB in Wakayama City as well as in the rest of Wakayama Pref. and in the whole country was highest in the age-group above 70 years of age, though the rate of Wakayama City was significantly higher (146.2 per 100,000 population) than that in the rest of Wakayama Pref. (98.5 per 100,000 population) and that in the whole country (90.3 per 100,000 population). Furthermore, the incidence rate of cases diagnosed as TB without bacteriological proof in Wakayama City (57.1 per 100,000 population) was significantly higher than that of the whole country (33.7 per 100,000 population). Therefore, we concluded that one of the causes of high incidence of TB in Wakayama City was due to inappropriate method of diagnosing TB. More extensive use of sputum examination and strict evaluation of cases without bacteriological proof are desirable to increase the accuracy of TB diagnosis in Wakayama City. PMID- 11398329 TI - [A case of drug-resistant pulmonary tuberculosis treated successfully following disappearance of rifampicin resistance after 17 years' chemotherapy]. AB - A female who first acquired pulmonary tuberculosis in 1962 when she was 25 years old, admitted to the National Hiroshima Hospital in 1982. Her sputum has been smear positive for acid-fast bacilli for 3 years before admission in spite of continuous antituberculous chemotherapy, and were resistant to isoniazid (INH) and rifampicin (RFP). She was treated with a regimen containing ethambutol (EB), prothionamide (TH) and enviomycin (EVM) but continued to be culture positive. Though she was treated with various regimens which include one to three sensitive drugs, her sputum continued to be positive for M. tuberculosis in the following 14 years. During the course, resistance to EB, TH, cycloserine (CS) and streptomycin (SM) emerged. Resistance to RFP temporarily retracted in 1988, but her sputum was bacilli negative only for 2 months after the addition of RFP to previous regimen, and followed by resurgence of RFP resistance. In 1992, data of drug sensitivity tests showed sensitivity to TH, CS and RFP in turn, which were not used for 3 to 5 years. In 1993, she was treated with RFP, TH and EVM successfully and continued to be bacteriologically negative for 7 years so far. Drug resistance to M. tuberculosis is induced by inappropriate chemotherapy as seen in this case. Regimens with less than three drugs without RFP and INH was not only insufficient to get cure but, what was worse, also induced additional resistance to used drugs. The reason of successful chemotherapy in this case was spontaneous disappearance of drug resistance to RFP and TH. This case suggests that the disappearance of drug resistance is possible, when drugs are not used for more than a few years, hence the successful treatment could be expected. However it must be emphasized that the drug resistance is produced by incorrect treatment as seen in this case, and its prevention is of the prime importance. PMID- 11398330 TI - [Where do we stand with BCG vaccination programme in Japan?]. AB - The current BCG vaccination program of Japan is critically discussed based on recent knowledge, especially with regard to its epidemiological aspects, in order to put the problem into perspective for Japan's future tuberculosis control program. 1. EFFICACY AND OVERALL EFFECTIVENESS: Various indicators of BCG efficacy have been proposed, and the meticulous analysis on the variability and the quality of these indicators seems to have formed a consensus on the efficacy, as seen in the recent meta-analysis studies. However, much has been left unanswered concerning the conditions under which the efficacy is guaranteed. The impact of the vaccination program on the population should also be considered in order to make decisions on the program. Comparing the age-specific tuberculosis notification rate between Japan and the USA, where there has been no BCG vaccination program, shows that the rate for 0 to 4 year olds is clearly lower in Japan than in the USA, while it is more than five times higher for all ages in Japan than in the USA. The statistics for Japanese children are superior to those of US children with respect to the speed of decline in notification rate as well. These observations support the overall effectiveness of BCG vaccination in Japan. 2. MECHANISMS OF BCG VACCINATION EFFICACY AND ITS DURATION: Two possible mechanisms of how BCG works to prevent tuberculosis were proposed. Epidemiological models based on each mechanism were subsequently tested by simulating 20 years' development of cases in the BCG vaccination trial by BMRC. In mechanism 1, the BCG-induced immunity is assumed to boost TB immunity in inhibiting the clinical breakdown of tuberculosis during the 10 to 15 years after the vaccination. In mechanism 2, the immunity makes the infection process abort (presumably, at 90%, during the seven years after infection, for example), leading to a smaller risk of future clinical development. So far, most epidemiological models implicitly assume mechanism 1 above. In animal experimental models, however, it has been difficult to simulate the mechanisms differentially, which has been one of the drawbacks to this argument. 3. EFFECTIVENESS OF REVACCINATION: Revaccination with BCG vaccine aims to restore or to endow immunological resistance through repeating vaccination to those who have partially or totally lost the immunity acquired from the primary vaccination. Although some animal experiments support the efficacy of revaccination with BCG, studies in humans have been rare and the results are variable. The observation of Polish infants and schoolchildren is suggestive of the efficacy, but it is not randomized and of questionable value. The recent study of Malawi is a randomized trial. It demonstrated that BCG revaccination protects against leprosy, but does not protect significantly against tuberculosis. It is possible, however, that it does protect against tuberculous lymphadenitis. The two above-mentioned possible mechanisms of BCG immunity were applied to a model analysis of BCG revaccination. It was known that revaccination effectiveness is very limited under any assumption, given the current Japanese epidemiological situation of tuberculosis, so that the demerits due to revaccination, such as strong local reactions, must not be neglected but should be carefully considered. At the same time, we should remember that this model analysis assumes that the primary vaccination is given to new borns with advanced and uniform techniques, which is not always true, and revaccination may supplement the technical failure of the primary vaccination. 4. DECIDING ON THE TOTAL DISCONTINUATION OF BCG VACCINATION PROGRAMME, JAPAN: The recommendations of WHO or IUATLD on the discontinuation of the BCG vaccination program are just conventional ones and the theoretical reasonings is difficult to accept. After all, the decision making should depend on the lay decision makers' subjective judgment balancing benefit and loss in terms of costs and health incurred by the policy, as shown by Waaler and Rouillon. The current Japanese BCG vaccination program is very expensive, but brings about some, though very small, benefit. This balance was compared with that of Sweden around 1975, when the program was discontinued. The comparison clearly showed that the cost effectiveness of the program in Japan today in superior to that of Sweden in 1975. PMID- 11398331 TI - [Chemical sensitivity]. PMID- 11398332 TI - [Fifty years history of Japanese society of allergology]. PMID- 11398333 TI - [Long-term study of airborne allergic pollen count, C. Japonica and cupressaceae in Japan]. AB - We have investigated the distribution of airborne pollen at the different eleven points in Japan from 1987 to 1998 using gravity sampler. To clarify the characteristics causative pollen for Japanese cedar pollinosis, we examined annual change of pollen counts, dispersing period and geographical difference of C. japonica and Cupressaceae pollen. C. japonica pollen occupied much more in Central Japan and Cupressaceae in the west of Japan than the other area. In Hamamatsu City, both of pollen counts were most of all and we found a tendency that the more pollen counts the longer dispersing period. As they reported that at the starting day of pollen in this method some patients had already suffered from allergic symptoms, we considered pollen grains were dispersing in spite of being continuously captured. In these twelve years we found that patients with Japanese pollinosis are exposured by causative agents during about 100 days every spring. But we could not observe the trend of increasing pollen counts and earlier starting day because of global warming. Further more we found that the pollen counts of C. japonica in autumn was increasing since 1994. As one of the factors of increasing patients with pollinosis, we thought that total exposure period of causative pollen every year were longer than that of 1980s. PMID- 11398334 TI - [Effect of fluticasone propionate at half dose of beclomethasone dipropionate divided twice daily and once daily in asthmatics inhaling beclomethasone dipropionate 800 micrograms daily or more]. AB - We conducted a 12 weeks' single-arm prospective study comparing beclomethasone dipropionate (BDP), 1/2 doses of fluticasone propionate (FP) twice daily and the same dose of FP once daily in 47 asthmatics who had been inhaling BDP 800 mcg daily or more. Peak expiratory flow (PEF), FEV1, a symptom score and a frequency of beta 2 agonist were all significantly better in FP twice daily phase than BDP phase (329 vs. 306 L/min, 1.87 vs. 1.76 L, 3.6 vs. 6.0/week and 2.7 vs. 4.5 puffs/day, respectively). There was no significant difference in these endpoints between FP twice daily phase and FP once daily phase. FP twice daily produced better plumonary function and symptom relief than the double doses of BDP. Furthermore, FP twice daily could, at least in a short-term basis, safely be changed into the same doses of FP once daily. PMID- 11398335 TI - [Quality of life in patients with perennial allergic rhinitis: using the Japanese version of the SF-36 Health Status Questionnaire]. AB - Perennial allergic rhinitis may impair daily activities. However it isn't known how it affects a patient's self-perceived health status. In present study, QOL was measured by MOS Short Form 36 Health Survey (SF-36) for assessment of useful information for individualization of care. 252 patients with perennial allergic rhinitis who visited 14 hospitals and clinics from June to August 1999 were questioned, and 249 patients were engaged in this study as subjects. Each patient's background was investigated, and the factors which influenced HRQOL score were identified. In addition, comparisons with healthy subjects and subjects with seasonal allergic rhinitis were performed. It was shown that age, gender, and the number of comorbid conditions were significant contributions to HRQOL score. It was considered that nasal obstruction had a strong influence on SF-36 score compared to other symptoms of nasal allergy. In general, QOL scores were significantly depressed in the patients with perennial and seasonal allergic rhinitis compared with healthy subjects. The difference of scores between perennial allergic rhinitis were not significant, although the patients with seasonal allergic rhinitis showed low scores compare with perennial allergic rhinitis. PMID- 11398336 TI - [A case of sarcoidosis exacerbated during improvement of bronchial asthma]. PMID- 11398337 TI - [Successful therapy of cyclosporin A in a case with idiopathic interstitial pneumonia]. AB - Seventy-one-year-old woman was visited to our hospital because of dry cough and dyspnea on effort. Fine crackle was audible on both lower lung fields. Joints and skin were normal. Laboratory examination revealed elevation of serum LDH and CRP level. Both anti-nuclear antibody and Jo-1 antibody were negative. Blood gas analysis showed hypoxia after exercise. Chest X-ray film showed reticular shadow in both lower lung fields. Chest CT finding showed patchy area of ground glass attenuation, air-space consolidation, and reticular shadow. Scintigram showed diffuse uptake of Gallium-67 in both lung. Transbronchial biopsy specimen revealed alveolar wall thickness, lymphocyte infiltration and swelling of type II pneumocytes. Bronchoalveolar lavage fluid analysis revealed elevation of CD4/CD8 ratio. She was given a diagnosis of idiopathic interstitial pneumonia. Combination therapy of cyclosporin A and steroid was performed. After therapy, her chest CT findings and her data of pulmonary function test were improved. Then therapy of cyclosporin A was continued and dose of prednisolone was gradually decreased. After that, she was suffered from respiratory tract infection. After administration of antibiotics and cyclosporin A, she was getting well without acute exacerbation of interstitial pneumonia. Since then, she was treated with cyclosporin A only and her pulmonary function test data were gradually improved more. It suggests that cyclosporin A may be useful for the treatment of idiopathic interstitial pneumonia. PMID- 11398338 TI - [Effect of Bakumondo-to on increased sensitivity of the cough reflex in a Sjogren syndrome patient with interstitial lung disease]. AB - A 63-year-old woman had complained of a persistent dry cough. Comparison of chest roentgenograms obtained 3 months earlier and at hospital admission, revealed progressive infiltrative shadows in bilateral middle and lower lung fields and mild volume loss. Hematologic study showed polyclonal hyper-gamma-globulinemia, positive anti-nuclear antibodies, and positive antibodies to Ro (SS-A) antigens. Ophthalmologic study demonstrated keratoconjunctivitis sicca. Parotid sialography showed an apple tree sign. Histopathologic features of labial salivary gland biopsy revealed agglomeration of at least 50 mononuclear cells. Chest CT showed non-segmental dense opacity in outer zones of middle and lower lung fields. Lung function test revealed a restrictive pattern with low diffusion capacity. Blood gas analysis showed hypoxemia (PaO2: 74 Torr, PaCO2: 32 Torr). Bronchoalveolar lavage fluid disclosed increased lymphocytes, and histologic features of a transbronchial lung biopsy included thickening of alveolar walls, infiltration of lymphocytes in the interstitium, and foamy cells in alveolar spaces. Therefore, the patient was diagnosed with primary Sjogren syndrome with interstitial lung disease. Bakumondo-to (Mai men dong tang) was started for the troublesome dry cough. The cough was subjectively relieved, cough scores were decreased, QOL scores improved, and sensitivity of the cough reflex measured by inhalation of capsaicin improved, but the chest roentgenogram was unchanged. Steroid hormone therapy was added, and radiographic abnormalities improved subsequently. Bakumondo-to, a traditional Chinese blended medicine, has notable antitussive activity in bronchitic guinea-pigs and in patients with postinfectious cough. It was concluded that Bakunondo-to may provide effective antitussive activity in some cases of interstitial lung disease with increased sensitivity of the cough reflex. PMID- 11398339 TI - Drug use and social circumstances: a study among different target groups in northwest Bangladesh. AB - The use of illicit drugs has become an increasingly serious problem among various sections of the Bangladesh population. However, there has been almost no research on the use of illicit drugs by Bangladeshi people. This paper examines drug use patterns and the social circumstances within which drug-taking behaviour took place among a sample of different target groups. The study incorporated a semi structured qualitative interview and a structured quantitative questionnaire. Demographic and drug use information was collected from a total of 154 drug users. Approximately three-quarters of the subjects reported that they had used alcohol and ganja in the month prior to being interviewed. The findings revealed interesting differences between the groups. For example, the student group was found to use phensidyl syrup (67%) more often than those of other groups and did not differ significantly in the use of ganja. The use of heroin and sedatives was widespread, with nearly one-third of subjects (40%) reporting having injected these drugs. The majority (41%) of most recent drug use occurred in conjunction with a group of people. Drug use was associated with frustration, peer pressure, family problems and curiosity. Peer education programs are likely to be the most effective harm reduction approach among new drug users. The implications of these findings are discussed. PMID- 11398340 TI - Lifetime prevalence of alcohol drinking, cigarette smoking, and solvent inhalation among junior high school students in Japan: tradition and urbanization. AB - To estimate the lifetime prevalence of alcohol drinking, cigarette smoking, and solvent inhalation among general junior high school students in Chiba prefecture, the author surveyed 6,115 students enrolled in 14 junior high schools in 1992. The lifetime prevalence of alcohol drinking was 78.4% of male subjects, 72.8% of female subjects, and 75.6% of all subjects. The lifetime prevalence of cigarette smoking was 30.7%, 14.9%, and 22.9%, respectively. The lifetime prevalence of solvent inhalation was 2.5%, 1.2%, and 1.9%, respectively. The past-year prevalence of solvent inhalation was 1.8%, 0.9%, and 1.4%, respectively. Furthermore, to reveal the relationship between the prevalence of drug use and the regional characteristics, multiple regression analysis was performed, using 7 indices that seemed to represent the regional characteristics. The multiple regression analysis revealed the following: 1) that the lifetime prevalence of alcohol drinking and cigarette smoking had a strong standardized partial regression coefficient with the percentage of owner occupants (Mochiieritsu in Japanese) and 2) that the lifetime and the past-year prevalence of solvent inhalation had a strong standardized partial regression coefficient with the ratio of daytime population to nighttime population (Chuyakanjinkouhi in Japanese). The author considered that these coefficients represented the relationship between traditional lifestyle and urbanization and drug use among junior high school students in Japan. PMID- 11398341 TI - Acute cytotoxic effects of ethanol on cultured mouse myocardial cells in a monolayer--enzymatic, chronotropic and ultrastructural studies. AB - The cytotoxicity of ethanol was investigated by detecting the leakage of lactate dehydrogenase (LDH) from three types of cultured mouse cells--non-proliferative but spontaneously and synchronously contracting atrial and ventricular myocardial cells in monolayer sheets, and also proliferative non-muscle fibroblast-like cells--over a 24-hr period (1, 3, 6 and 24 hr) after exposure to ethanol (12.5, 50, 200 and 500 mM). The beat rate and ultrastructure were also investigated. Only a 24-hr exposure to 500 mM ethanol markedly increased LDH leakage from all three types of cells, and it made 20-50% of these cells dead. The rate of spontaneous contractions of ventricular cells transiently increased after a 1-hr exposure to 200 mM ethanol and a 3-hr exposure to 500 mM ethanol compared with the control, though that of atrial cells did not show any significant change but tended rather to dose-dependently cease contracting. Atrial cells exposed to 500 mM ethanol never contracted, while ventricular cells ceased contracting only at 24 hr. After a 24-hr exposure to 500 mM ethanol some cells were necrosed ultrastructually, and the surviving cells contained giant mitochondria being bizarre in shape and many lipid droplets. Thus, ethanol induced cytotoxic effects terminating in death in cultured mouse heart-derived cells at a concentration of 500 mM. However, this concentration of ethanol did not induce any fatal effects within a 6-hr period, and neither did ethanol concentrations of less than 200 mM over a 24-hr period, as confirmed by the continuance of spontaneous contractions and ultrastructural observations. PMID- 11398342 TI - [Classification of alcohol metabolizing enzymes and polymorphisms--specificity in Japanese]. AB - Multiple forms and gene loci of human alcohol dehydrogenase (ADH EC: 1.2.1.3) and aldehyde dehydrogenase (ALDH, EC: 1.2.1.3) in the major pathway of alcohol metabolism have been found and characterized in the last two decades. With the coenzyme NAD, these enzymes catalyze the reversible conversion of organic alcohols to ketones or aldehydes, and aldehyde to acetic acid. The ADH genes are mapped to chromosome 4p21-25, but the ALDH genes are localized at different chromosomes. The cytochrome P450 2E1 (CYP2E1) gene, which is mapped to chromosome 10q24.3-qter contributes also the conversion of ethanol to acetaldehyde. Genetic polymorphisms have been reported in these alcohol metabolizing enzymes. The metabolisms of alcohol and acetaldehyde in liver and blood after drinking alcohol are thought to be influenced by the interactive action of these enzymes. Amongst the five major classes of the ADH subunits (alpha, beta, gamma, pi, chi, sigma), beta and gamma subunits show genetic polymorphisms. Recently a new nomenclature for ALDH genes has been recommend based on divergent evolution and chromosomal mapping. Two major isoforms designated as cytosolic ALDH1 and mitochondrial ALDH2 can be distinguished by their electrophoretic and kinetic properties as well as by their subcellular localization. Mitochondrial ALDH2 is a major enzyme in the oxidation of acetaldehyde derived from ethanol metabolism. The catalytic deficiency of ALDH2 isozyme is responsible for flushing and other vasomotor symptoms caused by higher acetaldehyde levels after alcohol intake. So far, frequencies of the two alleles of ALDH2 in Mongoloid have been reported in the different population groups. The catalytic deficiency of ALDH2 is caused by a structural point mutation at amino acid position 487, where a substitution of Glu to Lys resulting from a transition of G (C) to A (T) at 1510 nucleotide from the initiation codon has occurred. Individuals deficient in ALDH2 activity refrain from excessive drinking of alcohol due to the aversive reactions, leading to protection against alcoholism. Prevalence of the ALDH2*1 allele is associated with alcoholism, and subsequent studies have confirmed the allelic association with alcoholism in different ethnic groups. The effects of polymorphisms of ADH2 and CYP2E1 remained controversial, even in the same ethnic group. Investigation of mutations for the transacting cis-element in promoter region of the ALDH2 gene will provide important information with respect to regulation of this gene. Transfection assays using the first 600 bp of the upstream nucleotide sequences indicated that a region from -75 to -120 was necessary for the ALDH2 gene expression, and especially NF-Y/CP1 binding site from -92 to -96 (CCAAT box) is important in the expression of the gene. A novel polymorphism due to the nucleotide replacement at -357 G to A was found in all the population groups. Alcoholism is thought to be a multifactorial disease with complex mode of inheritance in addition to psychological and social factors, and many studies of family, adoption and twins concerning alcoholism have revealed that hereditary factor is an important determinant for developing alcoholism. Genetic association studies have contributed to the identification of a number of genetic risk factors for the chronic diseases influenced by genetic disorders and environmental factors. PMID- 11398343 TI - [Magnetic resonance (MR) imaging of the shoulder, elbow, and wrist]. AB - Thorough knowledge of small structures in the joints is becoming important as the resolution of magnetic resonance (MR) imaging has improved during the past decade. The author discusses the MR anatomy and pathology of three representative structures in the joints of the upper extremities: the labral-ligamentous complex of the shoulder, ulnar collateral ligament(UCL) of the elbow, and triangular fibrocartilage complex(TFCC) of the wrist. The labral-ligamentous complex is composed of the anterior labrum and three glenohumeral ligaments. Because of their similar MR appearance, the labral sulcus is difficult to differentiate from traumatic detachment of the labrum, even with the aid of MR arthrography. Insertion of the UCL to the medial condyle in the growing elbow presents different MR appearances according to age. Acute and chronic UCL tears are commonly seen in elbows with medial tension overload and are well depicted with three-dimensional Fourier transform(3DFT) gradient-echo images. 3DFT images are the most suitable for observation of the TFCC as well. Since abnormally high signals of the articular disc suggestive of tear are often seen in asymptomatic subjects, MR imaging may not reliably be used to differentiate clinically significant tears from asymptomatic defects. In this review article, the author emphasizes developmental anatomy and normal variations in the interpretation of MR images. PMID- 11398344 TI - [Usefulness of SPECT images in helping radiologists understand brain diseases]. AB - Nuclear brain imaging is able to show functional abnormalities of lesions that are not detectable by CT and MR images. The diagnostic keys of nuclear-imaging in terms of clinical usefulness are its early detection of lesions and determination of the efficacy of drug and surgical therapies. In dementic patients, F-18 FDG brain images can be diagnosed as Alzheimer's disease 12 months earlier than is possible on CT and MRI images, and can provide information for effective drug therapy. O-15 water CBF images can predict the effect of Nicholin by assessing transient increases in cerebral blood flow (CBF), thereby facilitating improvement in higher brain functions such as orientation. In stroke patients, brain SPECT images with Tc-99m HMPAO can predict fatal cerebral hemorrhage caused by anti-thrombic therapy by showing the decrease in count ratio (count ratio of infarcted to contralateral area of < 0.34) in the acute phase and identifying disruption of the blood brain barrier by showing hyperfixation in the subacute phase. Brain SPECT with I-123 IMP can also identify "misery" perfused areas resulting from reduced CBF and decreased vasoreactivity in the chronic phase. This criterion is utilized for patient selection for extracranial/intracranial bypass surgery, because patients with areas of poor perfusion might be indicated for such surgery. Since nuclear medicine images can accurately select candidates for drug or surgical therapies, they will be beneficial in reducing Medicare costs as well as in enhancing patients' quality of life as a result of the successful treatment. With the advancement of technology, nuclear medicine units that can simultaneously obtain CT images and can combine functional with anatomical images will provide more useful information for the diagnosis of brain disease. PMID- 11398345 TI - [MR hydrography of the abdomen: technical consideration of data acquisition and future prospects for clinical applications]. AB - Recent advances in MR hydrography have enabled various clinical applications in the areas of abdominal and pelvic disorders, for example, MR cholangiopancreatography (MRCP), MR urography(MRU), and MR hydrography of the fetus. Although the two-dimensional(2D) single-slice method provides excellent information as to the global relationship between lesions and various anatomical structures, small lesions or detailed anatomical characteristics may not be visualized owing to the partial volume effect. Source images of the 2D multislice method are most suitable for the detection of small lesions and detailed evaluation of anatomical structures as "tomographic imaging", while the three dimensional(3D) method is useful in obtaining data sets for 3D imaging. MR hydrography is a promising method for the noninvasive evaluation of various abdominal disorders, and it has the potential to play new roles in various anatomical regions. However, knowledge of the proper indications is essential for successful clinical application. PMID- 11398346 TI - [Efficacy of helical CT in evaluating local tumor extent of breast cancer]. AB - PURPOSE: To clarify the diagnostic accuracy of helical CT (HCT) in the determination of local tumor extent of breast cancer. MATERIALS AND METHODS: One hundred forty consecutive patients with breast cancer, including 87 invasive ductal carcinomas without extensive intraductal components (EIC), 44 invasive ductal carcinomas with EIC, 2 non-invasive ductal carcinomas, and 7 invasive lobular carcinomas, were included in the study. Three-dimensional tumor diameter including whole extent was measured on HCT, and the amount of invasion to fat tissue, skin, pectoral muscle, and chest wall was estimated using a three-step scale. These results were then compared with the pathological findings. RESULTS: Breast cancers appeared as areas of high attenuation compared with the surrounding breast tissue in all patients. Tumor extent was correctly diagnosed by HCT to within a maximum difference of 1 cm in 88 patients (63%) and within 2 cm in 122 patients (87%). Sensitivity, specificity, and accuracy in diagnosing muscular invasion of breast cancer using HCT were 100%, 99%, and 99%, respectively. Sensitivity, specificity, and accuracy in diagnosing skin invasion of breast cancer using HCT were 84%, 93%, and 91%, respectively. CONCLUSION: HCT was able to visualize all of the tumors and detect the correct tumor extent in most patients. PMID- 11398347 TI - [Evaluation of diagnostic accuracy of CRT monitor display for personal computer in the detection of small lung nodules: with particular emphasis on comparison between JPEG and wavelet compression]. AB - PURPOSE: To compare observer performance on cathode-ray-tube(CRT) monitors for personal computers with that on conventional radiographs in the detection of small lung nodules. MATERIALS AND METHODS: Fifty-eight normal chest radiographs and 58 chest radiographs with a small lung nodule were selected. Ten radiologists examined the original conventional films on a viewbox and digitized (8 bit) uncompressed and compressed images of the same patient on a color CRT monitor with a matrix of 1,600 x 1,200, and rated the presence of lung nodules with a five-level scale of confidence. The methods of compression used in this study were the JPEG and wavelet methods, with compression ratios of 6:1 and 15:1. Results were analyzed by receiver operating characteristic methods. RESULTS: There was no significant difference between film and digitized uncompressed and compressed images obtained by the JPEG and wavelet methods with a compression ratio of 6:1. No statistically significant difference was detected between film and digitized image with wavelet compression at 15:1. However, detection was less accurate on digitized images with JPEG compression at 15:1. CONCLUSION: Digitized (8 bit) uncompressed and compressed images with a compression ratio of 6:1 are acceptable for the detection of small lung nodules. Digitized compressed images at a compression ratio of 15:1 are also acceptable when the wavelet method is used. PMID- 11398348 TI - [Assessment of intra and extrahepatic perfusion during hepatic artery infusion chemotherapy using slow-injected gadolinium-enhanced MR imaging through implanted catheter-port-system]. AB - Twenty-four studies of intra-arterially slow-injected gadolinium-enhanced MR imaging through an implanted catheter-port system (reservoir-MR) were carried out in 15 patients with liver tumor. The flow rate of gadolinium injection was 0.1 ml/sec and a total of 3 mL was injected. Six consecutive phases, each with an acquisition time of 14 seconds, were obtained every 30 seconds. In all studies, the signal intensity of the drug delivery portion became very high. Twenty-three of 24 studies showed intrahepatic perfusion in the first phase. The hepatic vein was enhanced at the first phase in 10 and the second phase in 14. The abdominal aorta was enhanced at the second phase in all 24 studies. The portal vein was enhanced at the first phase in 4, the second phase in 13, and the third phase in 7 studies. Both intra- and extrahepatic perfusion were more clearly demonstrated by reservoir-MRI than by digital subtraction angiography through an implanted catheter-port system (reservoir-DSA); however, morphological changes in the hepatic artery were better demonstrated by reservoir-DSA than by reservoir-MRI. PMID- 11398349 TI - [Ultra-slow infusion dynamic MRI using an infusion pump: a new method for the evaluation of drug distribution in arterial infusion chemotherapy]. AB - Ultra slow infusion dynamic MR imaging using an infusion pump(IP-MRI) was performed in six patients with metastatic liver tumor or unresectable gastric cancer to evaluate ant-cancer drug distribution. We used un implanted port for the infusion of Gd-DTPA by infusion pump. On IP-MRI, the speed of Gd-DTPA infusion was very slow (0.01 ml/sec) , the same as drug infusion at chemotherapy. The contrast enhancement of tumors was extremely clear. Therefore, IP-MRI was considered feasible for the evaluation of drug distribution. PMID- 11398350 TI - [NMDA receptors and their interacting proteins in the postsynaptic density]. AB - Glutamate receptors and their cytoplasmic interacting proteins are thought to play an important role in the process of excitatory synaptic transmission in the mammalian central nervous system. NMDA receptors are a subtype of ionotropic glutamate receptors and specifically interact with PDZ-domain containing proteins such as PSD-95. This interaction is important in the clustering of NMDA receptors on the postsynaptic membrane and the molecular complex containing NMDA receptors and PSD-95 is likely to be the core structure of the postsynaptic density. In the first part of this review, biological significance of the diversity of NMDA receptor subunits is discussed. In the second part, implications obtained from our recent analyses on the structural remodeling of the postsynaptic density are presented. PMID- 11398351 TI - [Induction of cellular process elongation in hepatic stellate cells cultured on interstitial collagen gel: intracellular signaling mechanism]. AB - Hepatic stellate cells (HSCs), located in the perisinusoidal space of Disse, extend long cellular processes, which surround the hepatic sinusoids. However, after primary culture and following subculture using ordinary polystyrene culture dishes, HSCs lost their cellular processes and exhibited myofibroblast-like phenotypes. HSCs displayed rounded shapes when cultured on Matrigel containing the basement membrane components. On the other hand, HSCs exhibited the elongated cellular processes when cultured on interstitial collagen gel. This process elongation was induced by integrin-binding and the subsequent intracellular signaling pathways including protein kinases, protein phosphatases, PI 3-kinases, small G proteins, and microtubule-associated protein (MAP) subtype, MAP2C, and finally resulted from the reorganization of microtubules. The cellular processes contained vitamin A and matrix metalloproteinase-1. Such an HSC culture system using extracellular matrix components would be useful to study HSC functions in vivo, such as retinoid metabolism and reorganization of extracellular matrix. PMID- 11398352 TI - [Expression and localization of cell growth factors in the salivary gland: a review]. AB - The salivary gland secretes not only digestive enzymes but also various cell growth factors. Especially in rodents, granular ducts which develop between striated and intercalated ducts, are known to secrete epidermal growth factor (EGF) and nerve growth factor (NGF). Out of newly discovered growth factors, we have examined hepatocyte growth factor (HGF), transforming growth factor-beta (TGF-beta), insulin-like growth factor I (IGF-I), and basic fibroblast growth factor (FGF-2) and have demonstrated their expression and localization in the rat submandibular gland. HGF and TGF-beta show a very similar distribution pattern to EGF, i.e., exclusive localization in the secretory granules of granular duct cells. These factors are suggested to function as i) exocrine salivary factors, or ii) endocrine factors after reabsorption. In contrast, FGF-2 shows a different pattern of cellular and subcellular localization from EGF and others. FGF-2 is localized in cytoplasm of striated and excretory duct cells and pillar cells in granular ducts, but not in secretory granules of granular duct cells. Since the receptor for FGF also present in the same cells as its ligand, FGF is suggested to function in the salivary gland as an autocrine/paracrine factor. The possible physiological roles of salivary growth factors in the digestive organs are discussed based on our own data and an extensive literature. PMID- 11398353 TI - Cytoarchitectonic study of the cerebral cortex in the horse brain--classification of the cortex area. AB - A cytoarchitectural classification of the horse cerebral cortex was done to determine whether functional locations exist or not such as found in the human cerebral cortex. Two adult horse brains were examined by Nissl and myelin stained methods. Six cell layers of the cerebral cortex were identified and classified into five types areas based on the cytoarchitectural organization: agranular, frontal, parietal, polar and granulous types. The agranular type was seen around the gyrus sylvius. In layer V of the agranular type, there were the many large cells that seemed like Betz cells. This type was recognized as motor area. The frontal type was seen in the lower and parietal parts of the cortex. The parietal type was seen in the rostral and occipital lobes. The polar type was seen in the frontal and dorsal-occipital sides. The granulous type was seen in the anterior lower and posterior-lateral part of the cortex. The occupational ratios of the five types, that is, agranular, frontal, parietal, polar and granulous, to the whole cerebral cortex, were calculated by three-dimensional analysis using computer system as 19.6, 27.0, 28.9, 13.6 and 10.9%, respectively. As a result of myelin stain, horizontal fibers were seen in layer IV of parietal and polar types which looked liked the line of Gennari, therefore this area was recognized as the primary visual area. PMID- 11398354 TI - [Configuration of the liver segment observed in the frontal section including the origin of the left portal trunk at the hepatic hilum]. AB - After preparing threparing the frontal section including the origin of the left portal trunk at the hepatic hilum, 60 human livers (35, entirely; 25, partly) were dissected to reveal segmental configuration and the supplying portal vein branches. We usually observed two combinations of segments, i.e., S2, 4, 5 and 8 or S2, 3, 4, 5 and 8, in the frontal section including the origin of the left portal trunk. However, S8 was sometimes absent in the section when S4 extended to the right and/or upper side. S2 was consistently located dorsal to S3 despite the fact that 11.7% of the specimens carried an unexpected configuration showing a "lower" S2 in combined with an "upper" S3 in the frontal section. The latter case was associated with specific S2 and S3 segmental branches maintaining horizontal courses along a common plane. S4, S5 and S8 were usually arranged from the ventral to the dorsal aspect in this order. Four types of ventral short branches originated at or near the primary portal divisions and supplied the hilar parenchyme adjacent to S4 and/or the anterior segment (S5 or S8). These ventral short branches tended to be associated with the variations of the primary division. Dissection of the liver after frontal section provided a better understanding of the segmental configuration rather than an approach from the hepatic hilum. PMID- 11398355 TI - [SEM study on the dorsal lingual surface of the nutria, Myocastor coypus]. AB - The dorsal lingual surface of prenatal and postnatal nutria was examined by scanning electron microscopy (SEM). Filiform (Fi), fungiform (Fu), foliate (Fo) and vallate papillae (Va) were observed. The Fi differed in morphology by their location on the tongue and could be classified into 3 types: 1) conical, 2) long, and 3) short filiform papillae. The Fu were scattered among the Fi. Many Fo were observed on the posterolateral regions of the tongue. There were 2 Va separating the anterior 2/3 from the posterior 1/3 of the tongue. The rudiments of Fu, Fo and Va were visible earlier than those of the Fi. PMID- 11398356 TI - Basic science of articular cartilage repair. AB - As the ability to understand the peculiarities of successful healing of articular cartilage defects moves forward, it becomes clear that this complex orthopaedic problem soon will be successfully addressed. A multidisciplinary approach, combining clinical experience, cogent biomaterial designs, new cell biologic processes, biomechanical assessment, and modern molecular biology, clearly is leading toward clinically acceptable, viable, and consistent articular cartilage regeneration. PMID- 11398357 TI - Imaging of osteochondral injuries. AB - Acute injuries can produce fragments consisting of cartilage alone or cartilage and underlying bone. A purely cartilaginous fragment creates no direct radiographic abnormalities, whereas one containing calcified cartilage and bone appears as a radiodensity. The advent and refinement of MR imaging have led to the detection of occult injuries of subchondral bone and cartilage that can escape identification on routine radiographic analysis. The available data appear to indicate a substantial role for MR imaging in the analysis of lesion stability in cases of osteochondritis dissecans; however, further studies are required to determine its advantages over other methods, in particular, arthroscopy. Indirect evidence of cartilage abnormality on MR images relates to the identification of fluid at the interface between the fragment and the parent bone. The MR imaging technique influences dramatically whether such fluid is identified. MR arthrography employing the intra-articular injection of gadolinium compounds can be advantageous in the delineation of the chondral surface and in the detection of intraarticular bodies. Optimally, MR imaging would allow direct analysis of the cartilage surface, and specific imaging sequences that are most suited to this analysis are still evolving. PMID- 11398358 TI - Osteochondral injuries. Clinical findings. AB - Osteochondral injuries are common, mainly affecting a young, active population. Failure to recognize these injuries can lead to long-term disability. A heightened awareness and understanding of the common mechanisms of injury will lead to early diagnosis and rapid recovery. This, along with improved treatment modalities, will keep any long-term disability to a minimum. PMID- 11398359 TI - The three critical components in the conservative treatment of juvenile osteochondritis dissecans (JOCD). Physician, parent, and child. AB - There has been explosive growth of intensive, year-round sports for children of both sexes, with a resulting high degree of family involvement and commitment. Consequently, a diagnosis of juvenile osteochondritis dissecans (JOCD) impacts parent and child on many different levels. Compliance is the key element when a conservative treatment plan for JOCD is appropriate. An informed physician understands the dynamics between the "compliance triad" of physician, parent, and child, and recognizes that compliance is a process with foreseeable stages and characteristic behaviors. PMID- 11398360 TI - Chondral and osteochondral injuries. Diagnosis and management. AB - Osteochondral lesions are relatively common and usually occur as a result of trauma. They often are unrecognized acutely and lead to osteochondral defects and eventually osteoarthritis. Detection of these lesions has been aided by bone scan, CT, and MR imaging. Acute osteochondral fragments can be replaced and internally fixed. Chronic osteochondral defects can be treated with several methods designed to stimulate healing by either fibrocartilage or healing by transplantation of bone and cartilage or cartilage alone. The goal of all treatment methods is to provide a stable, congruent joint surface, restore function, and prevent the evolution of osteoarthritis in the injured joint. PMID- 11398361 TI - Treatment algorithm for osteochondral injuries of the knee. AB - The treatment of osteochondral fractures and OCD lesions in the knee is controversial. Many new procedures and techniques have been developed recently to address osteochondral lesions, indicating that no single procedure is accepted universally. Our treatment algorithm is based on the age of the patient, skeletal maturity, and the presence of adequate subchondral bone attached to the chondral lesion. Most nondisplaced lesions in the patient with open physes will heal with conservative treatment. The onset of skeletal maturity indicates a need for a more aggressive treatment approach. If adequate cortical bone is attached to the fragment, drilling of stable lesions, or drilling with fixation of unstable or loose fragments is appropriate. Autologous bone graft can be necessary to stimulate healing and properly reconstruct the subchondral bony contour. For failed fixation attempts or lesions not amenable to fixation, each treating surgeon must be proficient and comfortable with an articular surface reconstruction technique. The goal for the reconstructive procedure, to produce a smooth gliding articular surface of hyaline or hyaline-like cartilage, is possible using current techniques including mosaicplasty, osteochondral allograft transplantation, and autologous chondrocyte transplantation. Debridement, drilling, microfracture, and abrasion chondroplasty have been shown to result in fibrocartilage with inferior mechanical properties when compared with hyaline cartilage. No long-term studies have been published, however, to confirm the benefits of replacing osteochondral defects with hyaline cartilage rather than fibrocartilage. Although the results of many reconstructive procedures are quite encouraging with early follow up, the ultimate goal is to prevent long-term degenerative arthritis. Only well-designed prospective studies with long-term follow up will determine the adequacy of these procedures in reaching the ultimate goal. This treatment algorithm is based on the senior author's (WGC) experience with the complex dilemma of osteochondral lesions of the knee. PMID- 11398362 TI - Management of osteochondral injuries of the knee. AB - The management of articular cartilage lesions has yet to reveal a "right answer." Instead, it must be recognized as a multifactorial clinical challenge that requires the physician to consider surgical, biomechanical, and physiologic implications of the management chosen. The goal is to restore normal type II cartilage to the area of concern. Falling short of that, we must strive for the most reasonable of the facsimiles currently available. The science certainly will advance to assist our understanding of articular cartilage and the best way to approximate or replicate its properties. Continued research must examine which of the many variables are essential to address in contemplating these challenging cases. Basic science research appears to be the area of greatest promise. Perhaps elucidating the roles of Cartilage Derived Morphogenetic Proteins (CDMP) and other polyclonal stimulators of mesenchymal stem cells, and refining techniques of cartilage autotransplantation, should be included in the areas of focus. Studies evaluating stem cells as progenitors to cartilage-forming cells will bear watching. Long-term follow-up studies of all of the techniques reviewed are needed to give definitive answers about the durability of the repair and transplanted tissues. The orthopaedist taking care of these lesions is well served to have more than one option in managing these challenging clinical problems. PMID- 11398363 TI - Microfracture techniques in the treatment of osteochondral injuries. AB - Although many methods currently are available to treat articular cartilage lesions in the knee, microfracture has many inherent advantages. No other technique has been studied in the athletic population, and, in fact, most other procedures have been studied in the arthritic knee or in a mixed population of patients. Blevins et al have shown microfracture to be an effective tool in the treatment of chondral defects in both recreational and high-level athletes with mean follow-up of 3.7 years. Furthermore, microfracture is minimally invasive because it is arthroscopic through standard portals in most cases. The subchondral plate is preserved, unlike in other techniques, improving load bearing characteristics following healing. No heat necrosis or polishing is introduced into the subchondral bone and marrow with microfracture. The depth of penetration and location of puncture is controlled readily with the various awl angles. Surgical equipment and supply costs are minimal, without the need for expensive cell cultures or nonstandard equipment. No harvest site morbidity is present, unlike osteochondral, perichondral, periosteal, or chondral autograft procedures. Microfracture is not overly demanding technically, but emphasis must be placed on meticulous handling of the subchondral plate and surrounding healthy cartilage and adequate debridement of unhealthy cartilage. Finally, a good surgical technique is only as good as its rehabilitation. Strictly emphasizing the need for compliance with weightbearing restrictions and the use of CPM is essential to successful patient rehabilitation. PMID- 11398364 TI - Technical considerations in periosteal grafting for osteochondral injuries. AB - Periosteum has been used clinically for biologic resurfacing arthroplasty in small series of patients for almost two decades. The author's own experience with this technique in multiple joints, including the knee, has been similar to that already reported in the literature. Observations and considerations are discussed that might help avoid failure in future applications of this technique. Indications and surgical technique, including graft procurement and fixation, and postoperative treatment and possible complications are also described. The rationale for using periosteum as a chondrogenic tissue and the factors affecting its cartilage production are also outlined. PMID- 11398365 TI - Treatment of osteochondral injuries. Genetic engineering. AB - Articular cartilage injuries are commonly encountered problems in sports medicine and orthopaedics. The treatment of chondral and osteochondral lesions, which possess only a very limited potential for healing, still represents a great challenge to clinicians and to scientists. Experimental investigations reported over the last 20 years have shown that a variety of methods, including implantation of periosteum, perichondrium, artificial matrices, growth factors, and transplanted cells, can stimulate formation of new cartilage. Genetic engineering--a combination of gene transfer techniques and tissue engineering- will facilitate new approaches to the treatment of articular cartilage injuries. PMID- 11398367 TI - [Chronic urticaria and angioedema]. PMID- 11398366 TI - [New concepts about atopic dermatitis]. AB - The atopic dermatitis is a chronic inflammatory skin illness, with remissions and exacerbations, itch, and association with allergic rhinitis and asthma. There is a complex interrelationship of genetic, environmental, pharmacological and psychological factors that contribute to the development and severity of the illness: Different manifestations of immunological disorders are an increment in the number of IgE antibodies toward common antigens, an increment in the liberation of proinflammatory mediators by basophils and mast cells, peripheral and local eosinophilia, biphasic activity Th1/Th2 with the liberation of cytokines (IL-4, IL-5, IL-13), GM-CSF and the IFN-gamma caused by the cells Th1. an increment in the liberation of major basic protein, eosinophil cationic protein besides the expression of chemotactic factors by the monocytes (RANTES, eotaxin, vasoactive intestinal peptide, etc.). In 1980, Hanifin and Rajka made public the diagnostic criteria for the atopic dermatitis and it has been universally accepted as an standard for the diagnosis. Leung reported that a knowledge about the immunopathological bases of the atopic dermatitis has important clinical implications for the diagnosis and possible treatment there are multiple choices for a treatment because of the complexity of the illness. Among these are thalidomide and transfer factor as an immunomodulator treatment with acceptable safety and clinical efficacy. PMID- 11398368 TI - Correlation between respiratory function tests and evolution time in asthmatic patients. AB - A transversal and prospective study was performed to demonstrate a relationship between the evolution time of asthma episodes and alterations observed in the respiratory function tests (RFT) during asymptomatic periods. Asthmatic patients (n = 80) of both sexes, were studied, we investigated the evolution time of the asthmatic episodes and performed RFTs in the patients during their asymptomatic periods. Respiratory patterns were classified as normal, obstructive, or mixed (obstructive-restrictive), and a Spearman correlation test was performed. Twenty nine patients were male and fifty one female. All were between 5 and 49 years of age. Of the total number of patients, 13.7% fell into a normal pattern, 57.5% into an obstructive pattern and 28.7% in a mixed pattern. In the groups showing the shortest evolution time, the obstructive pattern was more common (75% of patients with less than 5 years of evolution time and 53.8% with an evolution time between five and ten years). The mixed pattern was more common in patients with more years suffering asthma (16.6% in the group of patients having 5 or less years of evolution and 50% in the group with more than 20 years). We found a Spearman value of 0.7, and we can conclude that there is more pulmonary damage associated to a longer evolution period of bronchial asthma. PMID- 11398369 TI - [Mycotic allergic sinusitis. Monographic review of the literature]. AB - Mycotic allergic sinusitis is a unique form of chronic sinusitis characterized by the presence of nasal polyposis and non-invasive mycosis. The incidence is still unknown but it is suggested it may have a geographical variation. Characteristically it is found in young patients with a history of atopy. Based on clinical data and paraclinical studies, diagnosis suspicion criteria have been proposed though the definitive diagnosis is made by identifying mycotic hyphae in the nasal secretion culture or the pathology specimen. There is still no consensus in treatment though the medical-surgical combination has been proposed. The prognosis varies according to the response to treatment. PMID- 11398370 TI - [Hypersensitivity to medications]. AB - drug hypersensitivity is an adverse reaction provoked by an immunologic response due to the consumption of a drug, or else, by reaction to one of their metaboliteS. Currently, it is considered that 10% of general population has susceptibility to a drug hypersensitivity to a drug or biological product. These reactions obey a pattern of factors of the individual and the medicine, that define the degree of sensitivity to the drug, and according to their characteristics it is possible to develop any type of immunologic reaction (Gel & Coombs), Nevertheless, it should be established that still there are mechanisms of drug hypersensitivity of drugs that have not been totally known, so they are not integrated to this classification and they are catalogued as an undefined type of reaction. On the other hand, there are drugs that provoke drugs reactions by non-immunologic mechanisms corresponding to the drug's different aggressive effects. According to the indicated concepts, we will show some basic concepts of the diverse mechanisms of drug hypersensitivity immunologic and non immunological, thus as some therapeutic and diagnostic concepts needed for the management of drug hypersensitivity. PMID- 11398371 TI - [Sensitivity and specificity of a questionnaire about respiratory symptoms similar to asthma]. AB - INTRODUCTION: Questionnaires are an epidemiological instrument to detect patients with asthma. MATERIALS AND METHODS: Worldwide standardized questionnaires were selected, taking the most important points for making the diagnosis of asthma. A questionnaire was applied to outpatients who attend to pulmonary disease service. The answers were compared with a "gold standard" proposed, consisted in clinical screening and pulmonary function tests. STATISTICAL ANALYSIS: Kappa coefficient, sensitivity, specificity and predictive values were obtained, with intervals of confidence of 95%. RESULTS: The questionnaires were applied to 120 patients, 70.83% women, the answering percentage was 99%. Mean age was 32 years with a range of 24-40. In 62.5% patients there was already a previous diagnosis of asthma. Positive Familial history was present in 65.83, cough in 85%, dyspnea in 80.83%, sneezes in 53.33% and wheezes in 77.5%. The was of kappa coefficient for previous medical diagnosis 0.96%, followed by positive familial history, wheezes and cough in 0.90%. The greatest specificity was for wheezes, dyspnea and previous medical diagnosis. The greatest specificity for previous medical diagnosis was wheezes and sneezes. CONCLUSIONS: The use of questionnaires for respiratory symptoms similar to asthma can be an option to detect groups of high risk. PMID- 11398372 TI - Why should we believe 210Pb sediment geochronologies? PMID- 11398373 TI - Land use influence on 137Cs levels in perch (Perca fluviatilis L.) and roach (Rutilus rutilus L.). AB - The environmental influence on Chernobyl-derived 137Cs levels in perch (Perca fluviatilis L.) and roach (Rutilus rutilus L.) was revealed using partial least squares regression (PLS). The 53 environmental predictors used describe land use in catchment areas, various catchment area and lake characteristics, lake water chemistry, and fish stock composition. The study showed a profound effect of land use on the 137Cs levels in fish. Radiocaesium deposited on arable land was retained in the soil to a greater extent than was 137Cs deposited on wetlands, which more easily leached out to the lake ecosystems. The 137Cs deposition close to the lakes had a more pronounced effect on 137Cs levels in the fish than did more distant deposition. The radiocaesium bioavailability is mainly governed by lake water cation content, as hardwater lakes had significantly lower 137Cs content in fish. Resuspension of 137Cs contaminated sediments only had a limited influence on the observed levels in fish. PMID- 11398374 TI - Distribution of organic carbon, selected stable elements and artificial radionuclides among dissolved, colloidal and particulate phases in the Rhone River (France): preliminary results. AB - The behaviour of radionuclides discharged from nuclear facilities in the Rhone River depends on their distribution among the dissolved, colloidal and particulate phases. A large water sample was fractionated using sequential ultrafiltration. Size distributions of organic carbon, Fe, Al, Si, Ca, Mg, Cu, Zn, 137Cs, 60Co and 106Ru were obtained. Our results show that organic colloids account for 11% of the total organic carbon content. Approximately 20% of the dissolved (< 450 nm) Fe and Al are in colloidal classes. 137Cs is not significantly transferred by the colloidal phase while 25% of 60Co or 106Ru is associated with organic and inorganic colloids. PMID- 11398375 TI - Plutonium contamination from accidental release or simply fallout: study of soils at Palomares (Spain). AB - The activity concentration of plutonium in an environmental sample does not usually constitute sufficient information to determine if it is due only to fallout. Alpha and gamma spectrometry are used here conjointly in the study of environmental soil samples to distinguish between samples showing plutonium contamination due to fallout exclusively, and samples contaminated with plutonium from another source. The method was applied to soil samples collected in Palomares (Spain), where an accidental release of aerosols contaminated with plutonium occurred. The two contributions (fallout and accidentally released plutonium) were separated by means of the activity ratios between various radionuclides present in the samples analyzed. The fallout level was estimated from the 239 + 240Pu/137Cs activity ratio. For samples showing contamination due to the accident, the 238Pu/239 + 240Pu and 239Pu/240Pu activity ratios were also calculated to determine the grade of plutonium of this contamination. PMID- 11398376 TI - Bioaccumulation and distribution of plutonium in fish from Gdansk Bay. AB - In this paper, the results of 238Pu and 239 + 240Pu determinations in four representative species of Baltic fish collected in Gdansk Bay; flounder, herring, cod and sprat, are presented and discussed. The plutonium isotopes are amongst the more radiotoxic nuclides. In the marine environment, the highest concentrations of plutonium are found in the sediments, but the complex biogeochemical cycle of the element means that it is also found in all other compartments. The activities of the fish samples were measured using alpha spectrometry and the concentrations of plutonium 238Pu and 239 + 240Pu were estimated for particular organs and tissues and the whole body. The 239 + 240Pu concentrations for fish species were: flounder 0.94 mBq kg-1 w.w., herring 2.22 mBq kg-1 w.w., cod 2.35 mBq kg-1 w.w. and sprat 0.33 mBq kg-1 w.w. On the basis of the 238Pu/239 + 240Pu activity ratio in the organs and tissues, the proportion of Chernobyl-derived plutonium in the Baltic Sea was calculated. The lowest values of Chernobyl plutonium were accumulated in flounder stomach, herring skin, cod intestine, the highest in cod gills and skin. PMID- 11398377 TI - Radioactive contamination of wood and its products. AB - This paper presents research on radioactive contamination of the three most common kinds of wood in Croatia--beech, oak and fir as well as acorn. Gamma spectrometric measurements carried out on the samples of bark and wood of beech, oak, fir and acorn have shown radioactivity contents ranging from 1.6 +/- 0.1 to 37.3 +/- 0.5 Bq/kg from deposited 137Cs, whose concentrations in the soil of Croatia have increased after the Chernobyl accident. Measurements have also shown the radioactivity originating from 40K and 214Bi, which are part of the natural composition of the soil. The distribution of the radionuclides in wood has been discussed, as well as the impact of radioactive contamination of wood by the artificial radionuclide 137Cs upon the forest ecosystem. According to the corresponding model, it has been calculated that a 10 h daily stay in a typical family house increases the annual radiation dose of the population, due to the deposited 137Cs in the structure or furniture, by 343 microSv. PMID- 11398378 TI - Contamination of Austrian soil with caesium-137. AB - Austria ranks among the countries that have been most strongly affected by the Chernobyl fallout. The mean contamination with 137Cs is 21.0 kBq/m2, of which 18.7 kBq/m2 is due to the Chernobyl accident, whereas global fallout contributes 2.3 kBq/m2. Maximum values of total 137Cs contamination are nearly 200 kBq/m2. Total deposition of Chernobyl 137Cs on Austrian territory is 1.6 PBq or a fraction of around 2% of the 137Cs released from the reactor. 2115 measurements were used to draw the Austrian "caesium map". The geographical pattern of fallout distribution shows regional differences of contamination as high as 1:100. PMID- 11398379 TI - Qualitative estimates of soil disturbance in the vicinity of CANDU stations, utilizing measurements of 137Cs and 210Pb in soil cores. AB - Anthropogenically derived 14C has been used to trace recent carbon input in a study of carbon accumulation and turnover in Canadian soils. In order to do so, documentation of the undisturbed nature of the soil horizons sampled was of prime concern. Although all the sites chosen for coring were currently uncultivated, detailed information on long-term land usage was not available. To overcome this problem, 137Cs profiles were measured in all the cores used in the study. 210Pb measurements were also made in cases where total 137Cs deposition was lower than predicted. For some sample sites, the data obtained showed correlated losses of both radionuclides, indicating that land disturbance and/or erosion had indeed occurred in these areas over the past 50 years, hence invalidating the use of those cores for carbon cycling studies. In a few cases a marked lack of correlation between these two radionuclides has made it necessary to hypothesize that chemical, rather than physical, processes have been partially responsible for the observed anomalies. Since results of this nature raise doubts about the reliability of the 137Cs method for identification of land disturbance, further investigation is warranted. PMID- 11398380 TI - Prophylactic antibiotics and children. PMID- 11398381 TI - Dentistry Canada fund. PMID- 11398382 TI - Dentistry Canada Fund. PMID- 11398383 TI - Debating the science of occlusion. PMID- 11398384 TI - Ethics. PMID- 11398385 TI - Justifying Water Fluoridation's ethical violations. PMID- 11398386 TI - Complementary dentistry: a more comprehensive term. PMID- 11398387 TI - Open minds, not empty minds. PMID- 11398388 TI - Benign cementoblastoma: a case report. AB - The case of a 23-year-old with a benign cementoblastoma is presented. The clinicopathologic features, treatment and prognosis are discussed and a brief review of the literature is presented. Although this neoplasm is rare, the dental practitioner should be aware of the clinical features that will lead to its early diagnosis and treatment. PMID- 11398389 TI - An unusual case of talon cusp on geminated tooth. AB - A rare case of talon cusp on geminated permanent central incisor is described. These developmental anomalies cause clinical problems including unsightly dental appearance, occlusal interference, displacement of the affected tooth, attrition, periodontopathy, irritation of the tongue, loss of space and malocclusion. Clinical and radiographic characteristics of these anomalies and modes of treatment are presented. Recognition of this condition and early diagnosis are important to avoid complications. PMID- 11398390 TI - Canada-wide standards: a pollution prevention program for dental amalgam waste. AB - Dentistry and society have long recognized the benefits of using silver-based amalgams to restore and maintain the dental health of patients. However, recent studies by health and environment experts have shown that mercury is of great concern when it enters the biosphere as a contaminant. A rational approach to pollution prevention is mandatory. This article explains the relationship between mercury, particularly dental amalgam waste, and the environment and describes a new pollution prevention initiative intended to ensure that the dental community becomes part of the solution to this serious environmental health problem. PMID- 11398391 TI - Evidence-based dentistry: Part II. Searching for answers to clinical questions: how to use MEDLINE. AB - The ability to conduct efficient literature searches is fundamental to the practice of evidence-based dentistry. In the second part of this series on evidence-based dentistry, strategic literature search techniques are discussed. MEDLINE, because of its breadth, depth and continuous maintenance by the U.S. National Library of Medicine (NLM), is the best source of evidence for health care. Although there are many gateways to MEDLINE, this paper highlights the user friendly versions of MEDLINE offered free on the Internet by the NLM. The use of well-established search tactics and the unique features of the NLM sites facilitate rapid, effective literature searches. PMID- 11398392 TI - A general method to identify bacterial genes regulated by cell-to-cell signaling. PMID- 11398393 TI - Genetic and chemical tools for investigating signaling processes in biofilms. PMID- 11398394 TI - In situ quantification of gene transfer in biofilms. PMID- 11398395 TI - Transcriptional analysis of genes involved in Pseudomonas aeruginosa biofilms. PMID- 11398396 TI - First stages of biofilm formation: characterization and quantification of bacterial functions involved in colonization process. PMID- 11398397 TI - Phenotype characterization of genetically defined microorganisms and growth of bacteriophage in biofilms. AB - Phenotypic characterization will be a pivotal aspect of future research in understanding the biofilm mode of growth. We hope that the concepts and techniques presented in this chapter will benefit other investigators in this field. Although initial studies will necessarily involve monocultures, eventually mixed culture work will have to be performed to understand biofilm growth in the natural environment. As the study of biofilm-phage interactions is new, there is considerable fundamental work that needs to be addressed. Here, we anticipate that some phage are better adapted to growth in biofilms, some are adept in growing in mixed culture biofilms, and others are better adapted to infecting planktonic organisms. Whereas biofilms are now widely accepted as a fundamental aspect of microbial growth in nature, the field of phage ecology is quite new and an exciting challenge for the future. PMID- 11398398 TI - Biofilm formation as a developmental process. PMID- 11398399 TI - Methods to detect and analyze phenotypic variation in biofilm-forming Staphylococci. PMID- 11398400 TI - In vivo models to evaluate adhesion and biofilm formation by Staphylococcus epidermidis. PMID- 11398401 TI - Genetic and biochemical analysis of Staphylococcus epidermidis biofilm accumulation. PMID- 11398402 TI - In vitro methods to study staphylococcal biofilm formation. PMID- 11398403 TI - Efficient RNA isolation method for analysis of transcription in sessile Staphylococcus epidermidis biofilm cultures. PMID- 11398404 TI - Assessment of metabolic potential of biofilm-associated bacteria. PMID- 11398405 TI - Characterization of extracellular chitinolytic activity in biofilms. AB - Extracellular enzymes produced by bacterial biofilms tend to become an integral, permanent part of the biofilm/substratum system. Thus, characterizing extracellular enzyme activity is an essential component of understanding biofilm ecology. Methods have been presented for characterizing three aspects of extracellular enzyme activity in biofilms: promoter activity of the structural gene, local catalytic activity, and kinetics of collective substrate degradation. The abundance of intracellular transcript derived from a structural gene is only indirectly related to the magnitude of catalytic activity of the corresponding enzyme. This relationship may be particularly tenuous in the case of extracellular enzymes, which must be transported out of the cell in order to become active. Fluorogenic substrates that allow direct detection of an increasingly greater variety of enzyme activities are becoming available. There are technical problems, originating from surface roughness and intrinsic fluorescence, associated with microscopic examination of biofilms on natural materials. Thin films provide one option for acquiring data about biofilms colonizing relevant materials. PMID- 11398406 TI - Surface sensing, swarmer cell differentiation, and biofilm development. PMID- 11398407 TI - Whole genome DNA microarray expression analysis of biofilm development by Vibrio cholerae O1 E1 Tor. PMID- 11398408 TI - Isolation and biochemical characterization of extracellular polymeric substances from Pseudomonas aeruginosa. PMID- 11398409 TI - Approach to analyze interactions of microorganisms, hydrophobic substrates, and soil colloids leading to formation of composite biofilms, and to study initial events in microbiogeological processes. PMID- 11398410 TI - Extracellular polymers of microbial communities colonizing ancient limestone monuments. PMID- 11398411 TI - Studying phototrophic and heterotrophic microbial communities on stone monuments. PMID- 11398412 TI - Identification of archaea in objects of art by denaturing gradient gel electrophoresis analysis and shotgun cloning. PMID- 11398414 TI - A dot-blot assay for adhesive components relative to probiotics. PMID- 11398413 TI - Lactobacilli as vehicles for targeting antigens to mucosal tissues by surface exposition of foreign antigens. PMID- 11398415 TI - Understanding urogenital biofilms and potential impact of probiotics. PMID- 11398416 TI - Acylated homoserine lactone detection in Pseudomonas aeruginosa biofilms by radiolabel assay. AB - We describe the development of a new radioactive assay for acyl-HSL production by bacterial cultures. The assay is based on the uptake of radiolabeled methionine and conversion of the radiolabel into SAM. The radiolabeled SAM is then incorporated into acyl-HSL by an acyl-HSL synthase. This assay is faster than previously used bioassays and shows no bias for the detection of acyl-HSLs of a particular length or side chain substitution. Acyl-HSL production can be monitored over a wide range of growth conditions in liquid culture. This assay can also be used in conjunction with a tube biofilm reactor to monitor acyl-HSL production by biofilm cultures. Ultimately this assay will allow comparison of acyl-HSL production by cells subjected to a variety of physiological conditions. PMID- 11398417 TI - Surface characterization and adhesive properties of bifidobacteria. PMID- 11398418 TI - Genetic and phenotypic analysis of multicellular behavior in Salmonella typhimurium. PMID- 11398419 TI - Gene transfer in bacterial biofilms. PMID- 11398420 TI - Conversion to mucoidy in Pseudomonas aeruginosa infecting cystic fibrosis patients. PMID- 11398421 TI - Subtractive hybridization-based identification of genes uniquely expressed or hyperexpressed during biofilm growth. PMID- 11398422 TI - Biofilm-induced gene expression and gene transfer. PMID- 11398423 TI - Directed movement and surface-borne motility of Myxococcus and Pseudomonas. PMID- 11398424 TI - Continuous culture models to study pathogens in biofilms. PMID- 11398425 TI - Biofilms in unsaturated environments. PMID- 11398426 TI - Biodegradable organic matter measurement and bacterial regrowth in potable water. PMID- 11398427 TI - Development of Thiobacillus biofilms for metal recovery. PMID- 11398428 TI - Methods used to assess biofouling of material used in distribution and domestic water systems. PMID- 11398429 TI - Microbial interactions to intestinal mucosal models. PMID- 11398431 TI - Analysis of microbial structure and function of nitrifying biofilms. PMID- 11398430 TI - Monitoring bacterial growth activity in biofilms from laboratory flow chambers, plant rhizosphere, and animal intestine. PMID- 11398432 TI - Computational and experimental approaches to studying metal interactions with microbial biofilms. AB - The structural and compositional heterogeneity of biofilms poses unique problems in metal fate and transport. A starting point for quantitative understanding of biofilm-metal interactions is surface complexation theory, with roots in chemical equilibria and thermodynamics. This approach permits fitting of experimental data to a variety of mathematical models from which predictive parameters, such as K, may be extracted. Applications of more sophisticated fitting routines such as tableau (as in FITEQL) or spectra pK methods provide a better measure of the heterogeneity. There remain large theoretical and computational challenges, as there is ample evidence to suggest that the principle of additivity is problematic, owing to chemical interactions between individual sorbent phases within biofilms. And finally, the question of how bacterial metabolic activity is likely to influence metal uptake by biofilms adds yet another layer of complexity for future investigations. PMID- 11398433 TI - Microscopy methods to investigate structure of potable water biofilms. PMID- 11398434 TI - Two-photon excitation microscopy for analyses of biofilm processes. PMID- 11398435 TI - Measurements of softness of microbial cell surfaces. PMID- 11398436 TI - Application of atomic force microscopy to study initial events of bacterial adhesion. PMID- 11398437 TI - Study of biofilm within a packed-bed reactor by three-dimensional magnetic resonance imaging. PMID- 11398438 TI - Methods for studying bacterial biofilms associated with plants. PMID- 11398439 TI - Detachment, surface migration, and other dynamic behavior in bacterial biofilms revealed by digital time-lapse imaging. PMID- 11398440 TI - Methods for predicting diffusion coefficients in biofilms and cellular systems. PMID- 11398441 TI - Limiting-current-type microelectrodes for quantifying mass transport dynamics in biofilms. PMID- 11398442 TI - Development of a standardized antibiofilm test. PMID- 11398443 TI - The MBEC Assay System: multiple equivalent biofilms for antibiotic and biocide susceptibility testing. PMID- 11398444 TI - Assessment of antimicrobial activity against biofilms. PMID- 11398445 TI - Retrieval of biofilms from the oral cavity. AB - With the use of the removable stents or bonded enamel piece models with or without a continuous bacterial layer, many in vitro or in vivo studies can be initiated. For example, studies on salivary pellicle formation, surface characteristics of biomaterials as they affect plaque development, antiplaque agents, the dynamics of adhesion of bacteria, interspecies adhesion of bacteria, the colonization of bacteria, the dynamics of bacterial growth in vivo, and the succession of growth in older supragingival plaques can be carried out. PMID- 11398446 TI - Biofilm acid/base physiology and gene expression in oral bacteria. AB - Environmental pH is one the major factors affecting the composition, biological activities, and pathogenic potential of the biofilms colonizing supragingival surfaces. In periodontal diseases, small changes in pH from the metabolism of amino acids and urea may influence the activity of proteolytic enzymes of host and bacterial origin. Still, there is a significant void in the understanding of pH-dependent gene expression in bacteria, in general, and this is of course a more acute problem when one considers there is virtually no information about gene expression in response to pH in biofilms. The development of new methods and applications of some of the techniques detailed above should help to ameliorate this situation and to generate much-needed data about the role of pH in biofilm composition, stability, and activity. PMID- 11398447 TI - In vitro modeling of biofouling of dental composite materials. PMID- 11398448 TI - An in vitro model for studying the contributions of the Streptococcus mutans glucan-binding protein A to biofilm structure. AB - The method described here for analyzing biofilms was sensitive enough to allow the detection of differences formed by pure cultures of S. mutans or a GbpA knockout strain. Other strains have also been tested, and the differences in biofilm structure were sometimes even more extensive (data not shown). The advantages of this method are that it is quick, inexpensive, and adaptable to almost any laboratory setting. The constant rotation of the cultures, which was employed to simulate salivary flow, appears to be a critical element for establishing biofilm differences. An analysis of protein profiles confirmed that the biofilm bacteria were metabolically distinct from the planktonic phase bacteria. For the strains tested, the variations in biofilm architecture could be visualized with or without magnification. Staining of the bacteria was not required, though we typically stained the biofilms with either crystal violet or Schiff's reagent. Altogether, this in vitro method for generating biofilms allowed the evaluation of visual, quantitative (confocal microscopy), and functional (antimicrobial susceptibility) differences. We have employed these methods in a reductionist approach to understanding the contribution of individual proteins to dental plaque development. These methods may also be useful in the screening of mutants that would be of greatest for testing in multispecies biofilms, animal models, or more complex biofilm models. PMID- 11398449 TI - Use of a continuous culture system linked to a modified Robbins device or flow cell to study attachment of bacteria to surfaces. PMID- 11398450 TI - Detection of streptococcal glucan-binding proteins in biofilms. PMID- 11398452 TI - Use of continuous culture bioreactors for the study of pathogens such as Campylobacter jejuni and Escherichia coli O157 in biofilms. PMID- 11398451 TI - Direct biofilm monitoring by a capacitance measurement probe in continuous culture chemostats. PMID- 11398453 TI - An open channel flow chamber for characterizing biofilm formation on biomaterial surfaces. PMID- 11398454 TI - Biofilms in flowing systems. PMID- 11398455 TI - Archaeal histones and nucleosomes. PMID- 11398456 TI - DNA binding proteins Sac7d and Sso7d from Sulfolobus. PMID- 11398457 TI - Reverse gyrases from bacteria and archaea. PMID- 11398458 TI - DNA gyrase from Thermotoga maritima. PMID- 11398459 TI - DNA topoisomerases VI from hyperthermophilic archaea. PMID- 11398461 TI - pGT5 replication initiator protein Rep75 from Pyrococcus abyssi. PMID- 11398460 TI - Topoisomerase V from Methanopyrus kandleri. PMID- 11398462 TI - Stability and manipulation of DNA at extreme temperatures. PMID- 11398463 TI - Ribonucleotide reductase from Pyrococcus furiosus. PMID- 11398465 TI - Ferredoxin from Thermotoga maritima. PMID- 11398464 TI - Preparation of components of archaeal transcription preinitiation complex. PMID- 11398466 TI - Methylguanine methyltransferase from Thermococcus kodakaraensis KOD1. PMID- 11398467 TI - DNA polymerases from euryarchaeota. PMID- 11398468 TI - RecA/Rad51 homolog from Thermococcus kodakaraensis KODI. PMID- 11398469 TI - Hyperthermophilic inteins. PMID- 11398470 TI - Assaying activity and assessing thermostability of hyperthermophilic enzymes. PMID- 11398471 TI - Chaperonin from Thermococcus kodakaraensis KOD1. PMID- 11398472 TI - Ferredoxin and related enzymes from Sulfolobus. PMID- 11398473 TI - Ferredoxin from Pyrococcus furiosus. PMID- 11398474 TI - Organic solutes from thermophiles and hyperthermophiles. PMID- 11398475 TI - Pressure effects on activity and stability of hyperthermophilic enzymes. PMID- 11398476 TI - Thermodynamic analysis of hyperthermostable oligomeric proteins. PMID- 11398477 TI - Dynamics and thermodynamics of hyperthermophilic proteins by hydrogen exchange. AB - The naturally occurring hydrogen exchange of protein molecules can provide nonperturbing site-resolved measurements of protein stability and flexibility and changes therein. The measurement and understanding of these issues is especially pertinent to studies of thermophilic proteins. This chapter briefly reviews the considerations necessary for measuring hydrogen exchange and translating HX measurements into these detailed protein parameters. PMID- 11398478 TI - Nuclear magnetic resonance analysis of hyperthermophile ferredoxins. PMID- 11398479 TI - Calorimetric analyses of hyperthermophile proteins. PMID- 11398480 TI - Ferredoxin:NADP oxidoreductase from Pyrococcus furiosus. PMID- 11398481 TI - Crystallographic analyses of hyperthermophilic proteins. PMID- 11398482 TI - Thermostability of proteins from Thermotoga maritima. PMID- 11398483 TI - Rubredoxin from Pyrococcus furiosus. PMID- 11398484 TI - Structural basis of thermostability in hyperthermophilic proteins, or "there's more than one way to skin a cat". PMID- 11398485 TI - NAD(P)H:rubredoxin oxidoreductase from Pyrococcus furiosus. PMID- 11398486 TI - Protein disulfide oxidoreductase from Pyrococcus furiosus: biochemical properties. PMID- 11398487 TI - Protein disulfide oxidoreductase from Pyrococcus furiosus: structural properties. PMID- 11398488 TI - DNA polymerases from hyperthermophiles. PMID- 11398489 TI - Medicaid/SCHIP 2000. PMID- 11398490 TI - HealthChoice--Maryland's Medicaid managed care experience. PMID- 11398491 TI - Medicare 2000. PMID- 11398492 TI - Mental health carve-outs: Part I. PMID- 11398493 TI - A review of Maryland insurance law. PMID- 11398494 TI - Health care coverage in Maryland. PMID- 11398495 TI - How I lost my eye. PMID- 11398496 TI - The nursing shortage in Maryland. PMID- 11398497 TI - Public health crises in Baltimore. Parallels between the Yellow Fever epidemic of the late 1700s and the spread of AIDS in the 1980s. PMID- 11398498 TI - Taxation with representation isn't so great either. PMID- 11398499 TI - Managed Care 2000: the year in review. PMID- 11398500 TI - [Value of 10 MHz ultrasound in diagnosis and follow-up of retrobulbar mucocele]. AB - Immediately after a flight from Europe to the USA a 45-year old German woman noticed an unilateral protrusion of the eye with retrobulbar pressure, diplopia and blurred vision. Except for a nasal polypectomy 23 years ago the past medical history was unremarkable. The above mentioned symptoms disappeared spontaneously but reoccurred after a second flight to Turkey six months later. The ultrasound examination of the orbital cavity showed a large suprabulbar mucocele which displaced the eyeball downwards. An intracerebral expansion was excluded by NMR tomography. The mucocele was removed surgically but reappeared when the patient had a cold. All symptoms disappeared within 2 weeks of taking antibiotics. Fluctuation of the air pressure in the aeroplane and below atmosphere pressure let the air in the sinuses expand. Via a connection between the nasal sinuses and the mucocele, the mucus is forcibly pressed into the mucocele, and the internal pressure rises. The clinical symptoms will therefore increase suddenly during a flight. The ultrasound examination is a reliable method for the differential diagnosis of retrobulbar tumor and should be used for follow-up examinations because of the simplicity and safety of the method. PMID- 11398501 TI - [Economics, quality and interdisciplinary cooperation in ultrasound diagnosis]. PMID- 11398502 TI - [Perivascular tissue vibrations: a criterium for high-grade stenoses of the internal carotid artery]. AB - Perivascular tissue vibrations, an artefact of the colour Doppler method, are already known to be a sonographic sign of stenosis but have not yet been investigated systematically in the examination of the internal carotid artery. AIM: The present study is intended to assess the value of this artefact in the diagnosis of stenosis of the internal carotid artery. METHOD: A total of 167 consecutive cases including a stenosis of the internal carotid artery of at least 50% were diagnosed according to the usual haemodynamic criteria using Doppler and duplex sonography and classified into 10% categories. Intermediate groups were formed for findings that could not be assigned unambiguously. By means of suitable apparatus settings each finding was examined to test whether a perivascular colour artefact was detectable. RESULTS: In the stenosis categories of 50% to 70% no perivascular colour artefact was found, whereas in the stenosis categories of 80% to 90% this was the case in 80% of the findings, and in the intermediate 75% category in 42% of the findings. The perivascular colour artefact was recognizable even under poor sonographic conditions. CONCLUSION: When a perivascular colour artefact is seen the diagnosis of a high-degree stenosis (or an AV fistula) is certain. The phenomenon is particularly valuable as a supplementary criterion of stenosis in cases of unfavourable sonographic conditions. However, the perivascular colour artefact may only be used to confirm, but never to exclude, a high-degree stenosis. PMID- 11398503 TI - [Doppler ultrasound diagnosis of increased intracranial pressure by comparison of 2 blood flow velocities in the extra- and intracranial segment of the internal carotid artery. 2: Findings in children with increased intracranial pressure]. AB - AIM: We investigated the correlation between intracranial pressure and flow parameters by simultaneous Doppler sonographic flow measurements in the internal carotid arteries and estimation of the intracranial pressure over the anterior fontanelle. METHOD: 12 infants (7 boys and 5 girls) with increased intracranial pressure were investigated. The corrected gestational age at investigation was 42 +/- 14 weeks, the weight was 3080 +/- 2540 g. Intracranial pressure was measured by applanation tonometry over the anterior fontanelle. The infants were subdivided into two groups: Group 1 (9 infants) with slightly increased intracranial pressure (16.9 +/- 3.3 cm H2O) and group 2 (3 infants) with moderately increased intracranial pressure (21.6 +/- 3.3 cm H2O). In all infants Doppler sonographic flow measurements within the intra- and extracranial segments of the internal carotid arteries were performed. From the flow profile, the peak systolic flow velocity Vs, the enddiastolic flow velocity Ved as well as the time average flow velocity TAV and the resistance index RI were measured. From the flow velocities, the I/E-ratio was calculated. The measured variables were compared with the values of matched pairs of a healthy control group of similar age and weight. RESULTS: Group 1 with slightly increased intracranial pressure displayed no significant difference of the RI within intracranial and extracranial sections of the internal carotid artery and the control group. The flow velocities, however, showed a significant increase within the intracranial segment of the internal carotid artery in comparison with the extracranial part of the internal carotid artery as well as the healthy control group. I/E-ratio for Vs increased significantly to 1.29 +/- 0.19, for Ved to 1.24 +/- 0.27 and for TAV to 1.08 +/- 0.27. Group 2 with moderately increased intracranial pressure displayed a significant increase of RI within the intracranial section of the internal carotid artery to 0.89 +/- 0.08 in comparison to the extracranial section with 0.79 +/- 0.07 and to the healthy control group with 0.75 +/- 0.07. All flow velocities decreased significantly within the intracranial section of the internal carotid artery in comparison to the extracranial part of the internal carotid artery and the healthy control group. The I/E-ratio was significantly reduced in group 2 with 0.76 +/- 0.11 for Vs, 0.38 +/- 0.21 for Ved, and 0.58 +/- 0.16 for TAV. CONCLUSION: Flow measurements in extra- and intracranial sections of the internal carotid arteries are an accurate method for semiquantitative estimation of increased intracranial pressure. This method is superior to the measurement of the RI. Slightly increased intracranial pressures below 20 cm H2O cause an increase of the I/E-ratio above 1, whereas the RI does not change. Moderately increase of the intracranial pressure above 20 cm H2O lowers the I/E-ratio significantly below normal values of 0.8, whereas the RI increases. PMID- 11398504 TI - [Use of 3-dimensional ultrasound for assessment of intrauterine device position]. AB - AIM OF THE STUDY: The purpose of our prospective study was to assess the accuracy of measuring and visualizing different types of IUDs with three dimensional ultrasound (3D) and two dimensional ultrasound (2D). METHODS: Ultrasound investigation was carried out in 147 patients 6 weeks after insertion of the IUD; in 127 cases a copper IUD was inserted, in 20 cases a Levonorgestrel-containing (LNG) IUD. Correct measurement was defined as the distance between top of the IUD and inner endometrium as well as IUD and myometrium (fundus) in the longitudinal plane (2D) plus the frontal plane (3D). The possibility to visualize the complete IUD was evaluated in the frontal plane and in the transparent mode after volume rendering. RESULTS: The distance measurement between copper IUD and myometrium or endometrium with 2D and 3D did not differ significantly (19.54 mm +/- 6.13 vs. 19.48 +/- 6.39, 5.58 +/- 4.57 vs. 5.21 +/- 4.57). There was no difference in the distance between LNG IUD and myometrium between the two ultrasound investigations (17.75 +/- 3.70 vs. 16.80 +/- 4.65), whereas the distance between LNG IUD and endometrium was significantly different (5.00 +/- 2.81 vs. 4.30 +/- 3.06, p = 0.03). The imaging of the LNG IUD was very clear with the 3D facilities, especially in the transparent mode. Furthermore, congenital anomalies of the uterus, like an arcuate uterus, were clearly detectable in the frontal plane (3D). CONCLUSION: Distance measurement can be obtained equal as well by 2D or 3D ultrasound. The 3D ultrasound offers the advantage of better visualization of LNG IUD and the assessment of anomalies of the uterine cavity. PMID- 11398505 TI - [Diagnosis of vesico-intestinal fistulas by contrast medium enhanced 3-D ultrasound]. AB - AIM: Standard diagnostic tools for vesico-intestinal fistulas are cystoscopy, cystography, colonoscopy, and contrast enema. The aim of our study was to evaluate the efficacy of transrectal 3D-ultrasound with contrast media in these patients. METHOD: From 5/98 to 12/99 we examined 10 patients with symptoms of a vesico-intestinal fistula (pneumaturia, faecaluria). After placement of a transurethral catheter a transabdominal ultrasound examination (Kretz Combison 530) was performed with the bladder half full to evaluate the bladder wall. Then the bladder was filled with diluted ultrasound contrast media (Levovist 40 mg/ml) to visualize the flow from the bladder towards the fistula. To verify a flow through the bladder wall a colour Doppler sonography of the region of interest was added. To evaluate form and extent of the fistula a transrectal ultrasound with 3D-image assessment was performed. RESULTS: Using this technique it was possible to demonstrate a vesico-intestinal fistula in 9 of 10 patients. In all cases these findings were confirmed by the standard diagnostic procedures. The fistulas were caused by: bladder carcinoma (n = 1), carcinoma of the colon (n = 2), Crohn's disease (n = 3) and diverticulitis of the sigma (n = 3). One patient presented with a neovesico-intestinal fistula in an irradiated local recurrence of bladder carcinoma. In one patient with Crohn's disease whose only symptom was pneumaturia all diagnostic tools failed to provide the diagnosis. CONCLUSION: For the first time vesico-intestinal fistulas could be demonstrated by ultrasound with 3D-image assessment using contrast media. This technique might be an effective addition to the standard diagnostics of vesico-intestinal fistulas reducing the exposure to radiation. PMID- 11398506 TI - [Value of ultrasound in diagnosis of bursopathies in the area of the Achilles tendon]. AB - AIM: The goal of this study was to check the usefulness of ultrasonography as a diagnostic tool for a bursitis in the area of the Achilles tendon. Furthermore, we assessed the shape and size of the retrocalcaneal bursa of normal subjects. METHOD: The ultrasonograms of 28 patients with a bursitis at the Achilles tendon were analysed. In addition, we sonographically examined the region of interest in 10 normal subjects (20 Achilles tendons). We used a 5 and/or a 7.5 MHz linear transducer. The examination was done in both standard planes. RESULTS: The examination of 10 normal subjects could not reliably demonstrate a bursa in the expected location of the subachillic bursa or the subcutaneous calcaneal bursa in a single case. We could, however, depict a subachillic bursitis (echofree to hypoechoic) in 22 patients. A subcutaneous calcaneal bursitis was found in 7 cases. CONCLUSIONS: The retrocalcaneal and subcutaneous calcaneal bursa is not demonstrable by ultrasonography in healthy people. Exsudation and proliferation of the bursa, however, facilitate the detection by ultrasound. Taking that into consideration, the sign of the bursitis is not the enlargement but the fact that the bursa is demonstrable at all by ultrasonography. In the case of the peritendinitis of the achilles tendon a concurrent bursitis should be looked for. During operative explorations an existing bursitis has to be removed. Ultrasonography is the method of choice for the diagnostic evaluation of a bursitis in the area of the achilles tendon. PMID- 11398507 TI - Clinical trial on the accuracy of a freehand and sensor-independent three dimensional power Doppler ultrasound system measuring diameters, volumes and vascularity of malignant primaries of the neck. AB - AIM: Diameters, volumes and vascularity of malignant primaries of the neck are of substantial importance for staging and prognosis. The purpose of this study was to determine the accuracy of a newly developed freehand and sensor-independent three-dimensional power Doppler ultrasound system (3D PDUS) in the assessment of these parameters. METHOD: 24 patients with squamous cell carcinomas of the neck underwent conventional ultrasound in B- and power Doppler-mode (US), 3D PDUS, and computed tomography (CT). Diameters (sagittal, longitudinal, and transverse) and volumes of the tumours were correlated with hand-segmentated CT images, whereas tumour vascularity was compared with US. RESULTS: 3D PDUS and CT correlated highly regarding diameters and volumes (correlation coefficients r = 0.98/P < 0.001 and r = 0.98/P < 0.001, respectively). 3D PDUS and US correlated highly regarding vascularity (r = 0.92/P < 0.001). CONCLUSION: Although 3D PDUS is based on a freehand and sensorless data acquisition, it appears to be as accurate as CT or US for measurements of diameters, volumes and vascularity of neck malignancies. In spite of the excellent results it has to be mentioned that this technique needs a lot of training and its general use has to be employed with caution. PMID- 11398508 TI - [Diagnosis of intramural bile duct varicose veins ("pseudo-cholangiocarcinoma sign"]. AB - We report the case of a patient with portal hypertension and portal vein thrombosis who was admitted to hospital for evaluation prior to a scheduled portosystemic shunt operation. Ultrasound examination revealed dilatation of intrahepatic bile ducts and echogenic thickening of the walls of both right and left main bile ducts as well as the common bile duct, highly suspicious of a carcinoma. Further evaluation, however, showed a varicosis of the bile duct walls to be the cause of the thickening of the walls, the varicosis resulting from the long-standing cavernous transformation of the portal vein and the portal hypertension. This tumour-like thickening of the walls, caused by numerous tiny varicose veins, is also known as "Pseudo-cholangiocarcinoma sign" in the Anglo American world. PMID- 11398509 TI - [Homocysteine and hyperhomocysteinemia]. AB - This paper reviews the biochemical and physiological data underlying hyperhomocysteinemia. Elevation of plasma homocysteine arises from disrupted of its metabolism (remethylation to methionine and transsulfuration to cystathionine) and is a function of a complex interaction between multiple genetic and environmental factors. Hyperhomocysteinemia, a conditions that recent epidemiological studies have shown to be associated with an increased risk of vascular diseases, may be a equal importance to hypercholesterolemia, hypertension and smoking in etiology of atherosclerosis. Based on the role of folic acid, vitamin B12 and B6 in homocysteine metabolism supplementation with these vitamins in treatment of hyperhomocysteinemia both in cardiovascular diseases and neural tube defects is reviewed. PMID- 11398510 TI - [Activation of acute phase proteins in patients with chronic hepatitis C treated with interferon-alpha 2a]. AB - The course of chronic viral hepatitis C (CVH C) depends in large degree from on non-specific immunological response. Interferon-alpha 2a (IFN-alpha 2a) affects immunological responses including acute phase proteins. The aim of the study was to determine whether chronic hepatitis C affects acute phase proteins and whether treatment with interferon-alpha restores disturbed acute phase proteins concentrations. Concentrations of acute phase proteins were compared with aminotransferases activity, a standard indicator of hepatocyte damage. We studied twenty patients (from 21 to 53 years old) with CHV C treated with interferon alpha 2a (6 million units, three subcutaneous injections a week). CRP, fibrinogen, C3 and C4 components of complement and albumin serum concentrations were determined before treatment, after 6 and 36 doses of interferon (after 2 and 12 weeks of therapy). Alanine aminotransferase activity was checked at the same time. No abnormalities in CRP, fibrinogen and albumin levels were observed before and during the treatment. It is regarded that low concentrations of C3 and C4 can lead to cryoglobulinemia, vasculitis, and polineuropathy. Both C3 and C4 were decreased among patients with CVH C. During therapy with interferon; levels of both proteins increased and achieved normal levels, especially among patients with effectiveness therapy of IFN-alpha. PMID- 11398511 TI - [Diagnostic value of testing the procoagulant activity of neoplasms in cases of stomach and esophageal neoplasms]. AB - The cancer procoagulant (CP) activity has been evaluated in homogenates of stomach and esophagus cancer tissues and in patients serum. Activity of CP in homogenates of stomach and esophagus cancer tissues as well as in serum of examined cancer patients were statistically higher than in the control group. The data indicate that estimation of CP activity in the neoplasmic tissues homogenates and in serum may be suggested as an new biochemical marker of cancer process useful in oncological diagnosis. PMID- 11398512 TI - [Evaluation of quality of life in patients with advanced colorectal cancer treated with topotecan]. AB - During last years a few new directions appeared in therapy of advanced colorectal cancer. Topotecan, a camptothecin analog, seems to be one of the promising novel drug in these cases. Its unique mechanism of action is connected with inhibition of the nuclear enzyme topoizomerase I. Assessing therapeutic effects of new cytotoxic drugs we should consider their impact on survival time and quality of life as well. The aim of our study was the attempt to assess the quality of life of patients with advanced colorectal cancer (IV stage in TNM scale), treated by topotecan. Clinical trial was performed in the group of 10 patients. Topotecan was administered intravenously at 1.5 mg/m2/day for 5 days and repeated every 21 days. Quality of life assessment was performed at special time points using Rotterdam Symptom Checklist. We observed improvement in quality of life in six out ten patients having advanced colorectal cancer. Two patients did not show any change in quality of life and two patients with progression of disease demonstrated lower quality of life during topotecan treatment. CONCLUSION: Topotecan has a positive influence on quality of life of patients with advanced colorectal cancer. Further study are needed to confirm this observation. PMID- 11398513 TI - [Cisplatin and vinorelbine therapy of previously treated advanced breast cancer (preliminary studies)]. AB - In this work it was investigated the efficacy of second and third line chemotherapy (vinorelbine, cisplatin/carboplatin) in patients with metastases breast cancer. The twenty pre-treated women with metastases to the liver, lung, lymph nodules and subcutaneous tissue were treated the regiment including navelbine 30 mg day 1 and 5 and cisplatin 100 mg/m2 day 1 every 3 weeks. The patients had been early received CMF and/or FAC. The total response rate was 40%. The median time to progression was 8 months. The stabilisation disease was at 20% patients. The tolerance of treatment was good and the frequent complication was leucopenia (WHO 2 and 3) and thrombocytopenia (WHO 1). The women received from 6 to 10 courses. PMID- 11398514 TI - [Evaluation of selected components in antioxidant systems of blood in patients with diabetes]. AB - Oxidative stress plays a major role in the development of chronic complications of diabetes. The aim of our study was to evaluate the selected components of the antioxidative system in well metabolically controlled diabetic patients. We also decided to assess the correlation between these parameters and duration of disease and the presence of it's late complications. The study was entered by 30 patients with type 1 diabetes (18 female and 12 male, aged 30.2 + 10.8 years with mean duration of disease 8.37 + 6.56 years, HbA1c 6.8 + 1.6%). 24 healthy, sex- and age-matched volunteers served as controls. We assessed the following parameters: reduced glutathione in erythrocyte lysate (colorimetric method by Bioxytech GSH-400), serum glutathione peroxidase (enzymatic immunological method by Bioxytech pl. GPx-EIA) and plasma superoxide dismutase activity (colorimetric method based on cytochrome c reduction). In comparison with controls, we found significantly higher reduced glutathione level (11.20 + 0.79 vs 3.92 + 0.62 mumol/l, p < 0.001) and markedly lower dismutase activity (27.49 + 1.32 vs 39.73 + 4.45 U/ml, p < 0.001). The levels of glutathione peroxidase did not differ significantly from values obtained in healthy subjects. We did not observe any correlation between the analysed parameters and duration of diabetes, HbA1c or presence of chronic complications of disease. The obtained results might indicate that antioxidative systems in the state of good metabolic control of diabetes have adaptive properties. PMID- 11398515 TI - [Digestive system diseases and style of nutrition among secondary school students]. AB - In the paper there was examined 246 pupils from secondary school. The auditorial survey including 23 questions was drawn up. The questions referred to digestive diseases occurrence among members of family, pupils under the survey, their style of nutrition, financial status of their family. It was interesting how kind of food, keeping regular hours, behaviour during eating is connected with frequency of digestive disease occurrence. 14.5% of pupils suffered from alimentary tract diseases especially constipation and peptic ulcer. A large percentage (46.2%) do not eat regularly, and 85% is in a hurry, doing something else during eating. 39.4% of examined use spices, ketchup, mustard every time, 17.5% do not eat vegetables, fruit at all, 45.9% eat sweets every day. Before leaving for school 45.9% pupils do not even eat breakfast. 69.1% of subjects claimed that stress is a reason of occurence their digestive disorders. In the paper abnormal style of nutrition is more frequent among pupils complaining of digestive system diseases. PMID- 11398516 TI - [Mast cells in chronic gastritis of children]. AB - Mast cell involvement in chronic gastritis in children was analyzed. 25 children with normal mucosa (controls), 20 children with chronic gastritis and 28 children with chronic gastritis and infected Helicobacter pylori were included to the study. Bioptic material from antrum and corpus of the stomach were stained with toluidine blue and anti-human mast cell tryptase to evaluate mast cell density. Changes in mast cells number were also estimated in 7 children before and after successful eradication of Helicobacter pylori infection. Mast cell density was significantly greater in children with chronic gastritis with or without Helicobacter pylori infection when compared to the controls. Mast cell degranulation was demonstrated by electron microscopy in children with chronic gastritis and infected Helicobacter pylori. Mast cell through it's numerous mediators may play a key role in chronic gastritis especially when Helicobacter pylori infection is present. PMID- 11398517 TI - [Use of three-dimensional reconstruction of the biliary tree in spiral computed tomography without using hepatotropic contrast media]. AB - The aim of the study was to assess the quality of biliary tract imaging without cholangiographic contrast medium and to evaluate the usefulness of 3-D reformations in determination of ductal anatomic relationship. The spiral 5 mm CT liver scans were performed after a bolus of 150 ml i.v. contrast agent in 15 patients with a malignant obstructive jaundice. The biliary tract was separated from surrounding hepatic parenchyma with segmentation computer algorithm and then 3-D models were produced. In all the patients, both biliary anatomy and the localization and degree of segmental obstruction were visualized with satisfying quality. The 3-D sCT models of dilated biliary tract can be obtained without cholangiographic contrast agent and can clearly depict ductal anatomy and pathology. PMID- 11398518 TI - [Influence of glucocorticoid steroid therapy on gastric and duodenal mucosa and Helicobacter pylori infection in children with nephrotic syndrome]. AB - The purpose of the study was to evaluate the influence of steroid therapy on endoscopic picture of gastric mucosa. The study comprised 22 children (17 boys and 5 girls) with the nephrotic syndrome aged 4-12 years (the mean 9 years) treated with prednisone. Children were divided into two groups taking into account duration of therapy. The group I (10 children) comprised patients with therapy duration not exceeding 3 months. The group II (12 children) comprised children treated 5 years in average. All patients underwent gastroscopic examination. Macro and microscopic lesions of gastric mucosa, their intensity and coexistence of Helicobacter pylori (HP) infection were estimated. CONCLUSIONS: 1. In nephrotic children during prednisone therapy different types of gastric and duodenal mucosal inflammatory changes occur. 2. Duration of prednisone therapy does not influence significantly on location, type and severity of mucosal inflammatory changes. 3. Frequency of coexistent HP infection increases with prednisone therapy duration. 4. Prednisone therapy of nephrotic children is indication for gastric and duodenal endoscopic examination. PMID- 11398519 TI - [Incidence of antibody detection against Toxocara canis and clinical symptoms in inhabitants of North-Eastern Poland]. AB - The purpose of this work was to evaluate the the level of exposition of humans on Toxocara eggs in North Eastern Poland on a base of serological tests and epidemiological and clinical data. 1025 inhabitants of North Eastern region of Poland were examine to detect antibodies against Toxocara antigens. 214 (20.7%) showed presence of antibodies against Toxocara. They were mostly males (71%) and country inhabitants (58.9%). No abnormalities in laboratory tests were found among these people. Some of them had complaints like joint pains (19.6%), temporary skin rash 2.3%). PMID- 11398520 TI - [Difficulties in diagnosis of Bourneville-Pringle's diseases]. AB - The authors present a case of Bourneville-Pringle disease in a 43 year-old female patient. The diagnosis was confirmed after 8 years from the beginning of disease. She was hospitalized several times and treated by different specialists. The diagnosis was confirmed in pulmonary department in SZChPiG in Bystra Sl. They observed multiorgan changes concerning not only the respiratory system, but also skin, kidneys, central nervous system. Medical history of patient, family data, the course of disease, histopathological changes in kidneys, specific ultrasonographic picture of the urinary system, high resolution computed tomography picture of lungs and classical computed tomography pictures of brain confirmed diagnosis. Tamoxifen was used in therapy. The improvement of quality of life and delay of natural progression was obtained. The course of disease observed during 20 years leads to ventilatory disturbances mainly obstructive type and respiratory insufficiency. The prognosis of presented patient still remains uncertain in spite of survival exceeding 20 years. PMID- 11398521 TI - [Trans isomers of unsaturated fatty acids--metabolic action and clinical effects]. AB - Fatty acids with the trans configuration are formed chiefly during hardening of oils and have been associated with increased risk of ischaemic heart disease. Dietary uptake of such fatty acids increases the level of lipoprotein (a) and LDL cholesterol, at the same time reducing the level of HDL cholesterol in plasma. As the content of saturated fatty acids in margarine is lower than in butter, the former product was for a long time considered as healthier. However, some margarines are rich in trans fatty acids. In other words, the risk of cardiovascular diseases can only be reduced by replacing butter with margarine checked for a low trans fatty acid content. The purpose of this paper was to review the literature dealing with the metabolic actions of trans isomers of unsaturated fatty acids and their clinical effects. PMID- 11398522 TI - [The role of transforming growth factors beta in pathogenesis of ulcerative colitis]. AB - Transforming growth factor beta is the protein playing a principal role in the intercellular signalling. The most important functions are: control of cellular growth, differentiation and migration. Moreover it stimulates synthesis of extracellular matrix proteins, modulates immune response and is responsible for angiogenesis, wound formation and tissue reconstruction. All these activities are involved in the development or healing of inflammatory bowel diseases. The role of transforming growth factors beta in the pathogenesis of ulcerative colitis are discussed in this paper. PMID- 11398523 TI - [Lipid metabolism. I. Role of insulin in lipid metabolism]. AB - The triacylglyceroles that comprise the bulk of lipids in the diet are hydrolyzed to free fatty acids, monoacylglyceroles and glycerol in the intestinal tract. During absorption through the intestinal tract mucosa, triacylglyceroles are resynthesized from free fatty acids, and glycerol-3-phosphate is formed in the intestinal mucose. these globules, called chylomicrons, pass through the liver and adipose tissue, they are reduced in size by an enzyme, lipoprotein lipase (LPL). In the postabsorptive period, free fatty acids and glycerol are released from adipocytes by neural and hormonal stimulation. The free fatty acids can be burned by almost all tissues of the body except the brain. They are burned in the mitochondria by a process of b-oxidation to acetyl-CoA, which can then enter the citrate acid cycle for conversion to CO2, adenosine triphosphate, and water. When excessive quantities of glucose are ingested, the glucose can be converted to a storage form, triacylglycerol. Fatty acids are synthesized by a series of reactions in which acetyl-CoA and malonylo-CoA residues sequentially condense until the fatty acid chain is completed. The fatty acids are then combined with glycerol-3-phosphate, generated in the liver, to form the neutral triacylglyceroles. The insulin has effects on both the synthetic (estrification) and breakdown (lipolysis) pathways. The promotion of triacylglycerol storage in fat is one of the most important of the actions of insulin. PMID- 11398524 TI - [Function of the growth hormone axis--insulin-like growth factors--insulin-like growth factor binding proteins in patients with chronic liver diseases]. AB - Up to date have demonstrated that more and more young people are affected by chronic liver diseased. There is a close relationship between the liver and endocrine system as far as hormone inactivation, synthesis of growth factors, proteins binding hormones and proteins binding growth factors in the liver are related. Impairment of the hepatocyte function may lead to disturbed homeostasis of the endocrine system. Current opinions on growth hormone (GH)--insulin-like growth factors (IGFs)--insulin-like growth factors binding proteins (IGFBPs) axis activity in men with chronic liver diseases are presented. PMID- 11398525 TI - [Diagnostic possibilities of anal endosonography in proctology. Part 1]. AB - On the basis of literature and own experience, the usefulness of endosonographic examination with the use of axial endoprobe was presented. The first part was devoted to the description of the endosonographic anatomy of normal rectum, examination technique and its usefulness in diagnosis of anal sphincters disorders. The endosonographic examinations have been performed in Poland for many years, mainly for prostate diagnostics with the use of multiplane transducers. We present a diagnostic possibilities of axial endoprobe of high frequency equipped with hard plastic cone and water balloon. PMID- 11398526 TI - [Tinnitus. I.--pathophysiology]. AB - Tinnitus is a sound impression in one or both ears appearing without any acoustic impulses from the environment. It can be a morbid symptom coming either from different parts of the auditory path (from the external auditory meatus to the auditory area in the cerebral cortex) or other organs. Usually ear noise is accompanied by hypacusis. According to British Tinnitus Association one of ten people suffers from ear noises and about 35-45% of adults have come down with it. This number is still increasing. The older the patients is the more frequent the noises are, especially among women after 40 and men after 50 years old. Because of varying origin different therapy methods are applied (surgical, pharmacological, psychotherapy). PMID- 11398527 TI - [Tinnitus. II.--treatment]. AB - Because of varying origin different therapy of tinnitus methods are applied (surgical, pharmacological, psychotherapy). Treatment of objective tinnitus of vascular origin is usually surgical. It is similar as in the case of tinnitus of mechanical origin. The procedure of operating. Treatment of subjective tinnitus is much more complicated because of difficulties with determining of its occurence. We can separate morbus Meniere, otosclerosis, which require precise treatment. Apart from these disease treatment of subjective tinnitus is multidirectional. It may include pharmacological treatment, psychotherapy, surgical treatment and other methods. PMID- 11398528 TI - [Let's calmly discuss death]. AB - Death is the only immovable issue in human's life and therefore it should be understood as human situation not as a tragedy. It is inseparable from life after all, and according to some theories it determines the essence of life, and partly it makes life possible. Moreover, it creates the feeling of responsibility for one's own existence. Most philosophers, except for the materialists, are in favour of the existence of the spiritual immortality. And consequently they reject the fear of the physical death, which relieves the sufferings, restores the calmness we were in before the nativity, and more importantly it secures equality as well as a fair treatment. Despite the existence of many philosophical concepts, death is and was an unknown matter. At present more than half the people die in hospital. The process of dying lasts much longer than before because of many medical and tending possibilities of maintaining life. The boundary between life and death became blurred. Furthermore, in many instances it is not life that is lengthened but dying. People are not accustomed to death and are afraid of it. The fear concerns not only the very moment of life end, but also many matters connected with death as well as with the passing person. Life lasting forever would be deprived of any meaning. One should consciously accept their own lot as well as the passing and they should be able to approve the inevitability of death. PMID- 11398529 TI - The Discipline: a pastoral care methodology with violent victims of violence. AB - Ours is a violent culture and violence creates victims. Many of these victims are themselves violent. This article describes The Discipline as used in the pastoral care process that seeks to rehabilitate these violent victims. The process involves 5 Rs: Rapport, Reflection, Realization, Reorientation, and Reintegration. PMID- 11398530 TI - The Dance with The Discipline. AB - The author became acquainted with The Discipline only recently and this article describes two pastoral visits in which The Discipline provides guidance for interactive presence. Thus this material represents how the author seeks to grasp and integrate it into her clinical practice in a short time. PMID- 11398531 TI - First steps: approaching The Discipline based, outcome oriented ministry model. AB - This article describes how a new staff chaplain is approaching and applying a discipline based, outcome oriented ministry model. He describes his move from being a lone, geriatric chaplain in a long term care facility to becoming a member of a team of staff chaplains in a teaching hospital who have disciplined themselves to an outcome oriented ministry model. The writer gives his perspective on The Discipline, and denotes ways in which his ministry has been impacted. Though the author has been involved with this process for only six months, both he and those in his care are experiencing some of the benefits of making tangible contributions and developing measurable outcomes in ministry. PMID- 11398532 TI - Playing well with others: The Discipline in a pediatric hospital. AB - This article contains a personal reflection on the way it feels, in fact and in theory, to move from a chaplaincy position in a clinical setting, to one where clinical chaplaincy is relatively new. The article presents the ways a clinically trained chaplain can introduce herself to a new setting, and how she must continually reevaluate her approach. PMID- 11398533 TI - Experience with clinical pastoral education in the context of The Discipline. AB - This article relates the experience of integrating The Discipline into the process and curriculum of an ACPE accredited clinical pastoral education (CPE) program involving residents and single unit students. It describes supervisor and student concerns as well as the conclusions they reached concerning the integration of The Discipline into The CPE process. PMID- 11398534 TI - The Discipline for doing spiritual care: variations on a theme. AB - This article describes the writers introduction to The Discipline and his decision to utilize selected aspects of it in direct care and in an educational milieu. The first section describes adaptation of the Profile (Concepts of Holy, Meaning, Hope, and Community) in the writer's work with addicted persons on a Recovery Center. The second section presents the employment of this Profile as a way to introduce CPE interns to the art of pastoral assessment. PMID- 11398535 TI - An adaptation of The Discipline in a clinical pastoral education program. Interview by Larry VandeCreek. AB - The author describes his perspective on providing pastoral care to hospital patients, his search for new ways to understand that ministry, his first contact with The Discipline, and his adaptation of it to the clinical and educational programs at his center. He discusses the need to help clinical pastoral education students to operationalize their ministry and to learn how to identify outcomes that are meaningful to health care decision makers. PMID- 11398536 TI - Using The Discipline for integrative ministry formation. AB - One United Methodist Annual Conference has begun a program of ministry formation education for probationary, first-appointment ministers in conjunction with Emory Clergy Care, Atlanta. This program uses an adaptation of The Discipline as an integral part of their ongoing formation education. Participants have learned the basics of planned pastoral care delivery, family and congregational systems thinking and self-supervision. Using The Discipline, they have been effectively adapting it to their local church settings and ministries. PMID- 11398537 TI - Prayer and healing: a chaplain's reflection on the dialogue with Larry Dossey. PMID- 11398538 TI - Living The Discipline on a stem cell transplant unit: spiritual care outcomes among bone marrow transplant survivors. AB - An oncology chaplain details, using The Discipline for Pastoral Care Giving, common themes in the spiritual journeys of stem cell/bone marrow transplantation survivors, and helpful chaplain interventions. PMID- 11398539 TI - A disciplined approach to spiritual care giving for adults living with cystic fibrosis. AB - Adults living with cystic fibrosis are less likely than other pulmonary patients to describe themselves as religious, to attend worship services regularly, to use god language, to describe their spiritual life, and in general, to give any obvious, outward indications of their spiritual strength, concerns, and depth. And yet, they have consistently demonstrated in chaplain-patient encounters an awareness of the function and importance of their spirituality in relation to life choices, coping with illness, facing mortality, and expressing life meaning, beliefs, and values. A disciplined approach by chaplains is a key component to engaging these patients so that each person's unique spiritual story unfolds. Results from The Discipline demonstrate how adults with cystic fibrosis are different in their expression and approach to spirituality from other pulmonary patients. PMID- 11398540 TI - The virtual desktop: the remote access solution. AB - The increasing expectation for easy, yet full remote access to library materials necessitates innovative thinking on the part of libraries. Other traditional methods of providing access don't always meet the needs of the library or the patrons. This paper describes the project of one library to develop a Virtual Desktop, making access easy to grant or withhold, and also making available a wider array of resources. PMID- 11398541 TI - Academic mentorship: an effective professional development strategy for medical reference librarians. AB - Academic mentorship is a professional development strategy that enables fledgling professionals to take advantage of the skills and expertise of the senior members for professional growth. Although widely practiced in many other professions, academic mentorship has not been widely reported in medical librarianship. Drawing upon personal experience, the author reports the success story of an academic mentorship program implemented in an academic medical library and argues for academic mentorship to be widely adopted in academic medical libraries. This paper first reviews the literature on the concept of mentoring in an academic setting, and then describes the background, rationale, methods, and results of the mentorship programs the author has experienced. Lastly, based upon an analysis of several surveys and studies on coping skills for quality job performance of health sciences reference librarians, the paper discusses mentorship as one effective means to ease a new medical reference librarian's transition from his/her pre-service experience to the professional world of medical librarianship. It calls on other health sciences librarians to consider developing their own mentorship programs to promote their professional development and personal growth. PMID- 11398542 TI - Introduction to resources in molecular genetics. AB - Perhaps no other discipline has experienced as dramatic an information explosion in recent years as has the field of molecular genetics. While the flood of scholarly publications stemming from this research has been extensively indexed in MEDLINE, a number of new databases have been developed that provide direct access to the primary sequence data as well as to curated, reference information. A select group of these databases, including NCBI's GenBank, LocusLink, RefSeq, OMIM, and Genes and Disease, is examined. Following a brief description of each database, a proposed instructional program for their integration into an undergraduate genetics course at the University of Vermont is described. PMID- 11398543 TI - TRIPin' on a new EBM resource. PMID- 11398544 TI - Patients' rights on the World Wide Web. AB - Managed care reform, commonly referred to as "patients' rights" legislation, has become a hot topic. Many groups, including consumers, health care professionals, employers, managed care organizations, political parties, and government agencies, have strong opinions about measures that should be taken and what the outcomes of these measures might be. Those investigating this multidisciplinary topic will want to examine health care administration, ethics, health services research, and political science sources. Web resources covered in this article include: clearinghouses; government agencies; federal legislative and legal sites; and home pages of professional and trade associations, policy research institutes, and consumer advocacy organizations. PMID- 11398545 TI - Helping seniors capture the power of the Internet. PMID- 11398546 TI - Collaborating to create an online evidence-based medicine tutorial. PMID- 11398547 TI - Mediated computer search services relative to instruction services: a survey of one health sciences library. AB - To assess the quality and usefulness of one health sciences library's mediated computer search service, a survey was undertaken to determine satisfaction rates, why users do or do not use the service, and how useful the service is perceived to be in comparison to instructional service. Satisfaction rates were high, with users indicating librarian expertise and time/cost savings as the main reasons for using the service. Non-users indicated that they preferred to do their own searching, and many were unaware of the service. Though a majority of respondents do not currently use the service, surprisingly a majority of respondents placed significant value on the mediated search service in relation to instruction. PMID- 11398548 TI - The mid-palatal suture in young adults. A radiological-histological investigation. AB - The aim of the present study was to analyse which histological-histomorphometric findings correspond to a radiologically diagnosed open (group I) or closed mid palatal suture (group II) on occlusal radiographs. For this purpose, 30 radiological regions of interest (rROI) from specimens obtained from 10 subjects ranging in age from 18 to 38 years were evaluated, and compared with the suture morphology, mean sutural width, and degree of suture closure on stained sections (3/rROI). The results showed that whether or not a radiologically visible suture can be classified as 'open' depends predominantly on the main oronasal suture course in relation to the X-ray path, rather than on factors such as suture width and degree of obliteration. The mean sutural width was 231 microns in group I (n = 10 rROI) and 201 mu in group II (n = 20 rROI). The degree of obliteration in relation to the total oronasal suture length was 0.45 in group I and 1.30 per cent in group II. No significant differences were found between the corresponding parameters of the two groups. The term 'suture obliteration' or 'fusion' should be avoided if a suture is radiologically not visible, since in 11 of the 20 rROI in which the suture was not visible no obliteration was recorded morphometrically. PMID- 11398549 TI - A prospective optical surface scanning and cephalometric assessment of the effect of functional appliances on the soft tissues. AB - The aim of this study was to evaluate the effect of different functional appliances on the soft tissues as assessed by cephalometry and optical surface scanning. Forty-two patients were randomly allocated to Bass, Twin Block (TB), and Twin Block + Headgear (TB + Hg) groups. Lateral cephalograms and optical surface scans were recorded before and after the 10-month study period. ANOVA was used to test the cephalometric variables for differences at the 5 per cent level. The optical surface scanning and cephalometric results were consistent in the sagittal dimension. In the vertical dimension, however, the optical surface scans consistently recorded a greater increase compared with cephalometric values. No differences were detected with regard to cephalometric values at the 5 per cent level. However, the Bass appliance produced greater forward positioning of soft tissue pogonion as assessed by optical surface scanning. PMID- 11398550 TI - Oral appliances for the management of severe snoring: a randomized controlled trial. AB - The aim of this randomized controlled trial was to assess the effectiveness of a mandibular advancement appliance (MAA) in managing severe snoring. Twenty-eight adults with severe snoring and normal overnight oximetry were recruited from sleep disorder clinics. A maxillary placebo appliance and a MAA were worn by each subject for a period of 4-6 weeks each. Questionnaires at baseline and after each appliance period assessed bed partners' reports of snoring severity (loudness and number of nights per week), and patients' records of daytime sleepiness. Twenty five subjects completed the entire trial. The MAA was significantly more effective than the placebo in reducing the frequency and loudness of snoring, the reported daytime sleepiness and the frequency of morning tiredness. Excessive salivation was the most commonly reported complication. It was concluded that the custom-made MAA was significantly more effective than the placebo in managing the main symptoms of severe snoring. However, not all subjects' partners reported an improvement with the MAA, with 84 per cent reporting a reduction in snoring loudness and 76 per cent reporting snoring on fewer nights per week. PMID- 11398551 TI - Orthodontic side-effects of mandibular advancement devices during treatment of snoring and sleep apnoea. AB - The aims of this study were to investigate possible orthodontic side-effects following the use of mandibular advancement devices (MAD) in adults with snoring and sleep apnoea. A second objective was to analyse the effect of the appliance design. Seventy-five patients treated with MAD and 17 reference patients were studied at follow-up after 2.5 +/- 0.5 years. In the test group, 47 patients were provided with soft elastomeric devices, while the remaining 28 patients received hard acrylic devices. The treatment induced a change in overjet of -0.4 +/- 0.8 mm (mean +/- SD) and a change in overbite of -0.4 +/- 0.7 mm (mean +/- SD). These changes were larger than those found in the reference group (P < 0.01). The odds ratio (OR) for the largest quartile of reduction in overjet was 3.8 in patients using hard acrylic devices compared with those using soft elastomeric devices (P < 0.05). A large reduction in overjet in patients using the hard acrylic devices was unrelated to the degree of mandibular protrusion by the device. The OR for a large reduction in overjet in patients using the soft elastomeric devices with a protrusion of 6 mm or above was 6.8 compared with smaller mandibular protrusions (P < 0.05). The results indicate that the orthodontic side-effects are small during the treatment of adult subjects with MAD for snoring and sleep apnoea, especially in patients using soft elastomeric devices with mandibular protrusions of less than 6 mm. The follow-up of patients treated with MAD is recommended, as individual patients may experience marked orthodontic side-effects. PMID- 11398552 TI - Congenital tooth anomalies and malocclusions: a genetic link? AB - The aim of the present study was to investigate putative relationships between different malocclusions such as Class III and Class II division 1, and congenital tooth anomalies. Two-hundred Class III and 215 Class II division 1 patients were examined for the presence of any of the following congenital tooth anomalies: maxillary incisor hypodontia, maxillary canine impaction, transpositions, supernumerary teeth, and tooth agenesis. Their occurrence rates were then calculated as a percentage of the total sample and were compared for statistical differences. The results revealed no statistical difference (P > 0.05) in the occurrence rates of upper lateral incisor agenesis, peg-shaped laterals, impacted canines, or supernumerary teeth between the Class III and the Class II division 1 malocclusions. When the occurrence rate of all congenital tooth anomalies was compared between the two malocclusions, Class III subjects showed significantly higher rates (P < 0.05). Comparison with published surveys on general populations showed similar occurrence rates. It can be concluded that subjects with Class III and Class II division 1 malocclusions show patterns of congenital tooth anomalies similar to those observed in the general population. Congenital tooth anomalies may represent another criterion for the study of malocclusion, with respect to their origin and development. PMID- 11398553 TI - Prevalence of malocclusion and orthodontic treatment need in children and adolescents in Bogota, Colombia. An epidemiological study related to different stages of dental development. AB - The aim of the study was to assess the prevalence of malocclusion in a population of Bogotanian children and adolescents in terms of different degrees of severity in relation to sex and specific stages of dental development, in order to evaluate the need for orthodontic treatment in this part of Colombia. A sample of 4724 children (5-17 years of age) was randomly selected from a population that attended the Dental Health Service; none had been orthodontically treated. Based on their dental stages the subjects were grouped into deciduous, early mixed, late mixed and permanent dentition. The registrations were performed according to a method by Bjork et al. (1964). The need for orthodontic treatment was evaluated according to an index used by the Swedish National Board of Health. The results showed that 88 per cent of the subjects had some type of anomaly, from mild to severe, half of them recorded as occlusal anomalies, one-third as space discrepancies, and one-fifth as dental anomalies. No clear sex differences were noted, except for maxillary overjet, spacing, tooth size (all more frequent in boys), and crowding (more frequent in girls). Occlusal anomalies and space discrepancies varied in the different dental developmental periods, as did tipped and rotated teeth. Little need for orthodontic treatment was found in 35 per cent and moderate need in 30 per cent. A great need was estimated in 20 per cent, comprising children with prenormal occlusion, maxillary overjet, or overbite (> 6 mm), posterior unilateral crossbite with midline deviation (> 2 mm), severe crowding or spacing, congenitally missing maxillary incisors, impacted maxillary canines or anterior open bite (> 3 mm in the permanent dentition). Urgent need for treatment was estimated to be 3 per cent, comprising subjects with extreme post- and pre-normal occlusion, impacted maxillary incisors or extensive aplasia. PMID- 11398554 TI - A comparison of information retention at an initial orthodontic consultation. AB - The exchange of information is an everyday part of orthodontic treatment. However, the amount of information that is understood and retained, by patients and their parents, is not known. There has been very little research in the area of information retention in dentistry. This has implications with the demands for improved provision of information for patients. This questionnaire-based study, compared the effectiveness of written, verbal, and visual methods of providing orthodontic information. It assessed the retention of this information, by patients and parents, in both the short- and long-term. Twenty-eight patients and their parents, were allocated alternately into one of three groups, receiving written, verbal, or visual information. Short-term retention of knowledge was assessed 10-15 minutes after receiving the information and long-term retention rated by a second questionnaire mailed 8 weeks later. Overall, little difference was found between the three methods. The findings suggested that verbal information should not be given to patients unless supplemented by written and/or visual information, and that parents were more attentive to verbal instructions than their children. PMID- 11398555 TI - Temporomandibular disorders in relation to craniofacial dimensions, head posture and bite force in children selected for orthodontic treatment. AB - The present study examined the associations between craniofacial dimensions, head posture, bite force, and symptoms and signs of temporomandibular disorders (TMD). The sample comprised 96 children (51F, 45M) aged 7-13 years, sequentially admitted for orthodontic treatment of malocclusions entailing health risks. Symptoms and signs of TMD were assessed by 37 variables describing the occurrence of headache and facial pain, clicking, jaw mobility, tenderness of muscles and joints, and the Helkimo Anamnestic and Dysfunction indices. Craniofacial dimensions (33 variables), and head and cervical posture (nine variables) were recorded from lateral cephalometric radiographs taken with the subject standing with the head in a standardized posture (mirror position). Dental arch widths were measured on plaster casts and bite force was measured at the first molars on each side by means of a pressure transducer. Associations were assessed by Spearman correlations and multiple stepwise logistic regression analyses. The magnitudes of the significant associations were generally low to moderate. On average, temporomandibular joint (TMJ) dysfunction was seen in connection with a marked forward inclination of the upper cervical spine and an increased craniocervical angulation, but no firm conclusion could be made regarding any particular craniofacial morphology in children with symptoms and signs of TMJ dysfunction. Muscle tenderness was associated with a 'long face' type of craniofacial morphology and a lower bite force. Headache was associated with a larger maxillary length and increased maxillary prognathism. A high score on Helkimo's Clinical Dysfunction Index was associated with smaller values of a number of vertical, horizontal, and transversal linear craniofacial dimensions and a lower bite force. PMID- 11398556 TI - Temporomandibular disorders in adults with repaired cleft lip and palate: a comparison with controls. AB - The purpose of this study was to investigate the prevalence of temporomandibular disorders (TMD), and assess psycho-social distress in adult subjects with repaired complete cleft lip and palate (CLP). Sixty-three adults (42 males and 21 females, mean age 24.2 years, range 19.5-29.2) with repaired CLP (CLP group) were compared with a group of 66 adults without cleft (non-cleft group, 49 males and 17 females, mean age 25.5 years, range 20.2-29.9). All subjects underwent a clinical TMD examination, which followed the guidelines in the Research Diagnostic Criteria for TMD (RDC/TMD). Jaw function was assessed by evaluating answers to the mandibular function impairment questionnaire (MFIQ). Tension-type headache was diagnosed according to the International Headache Society (IHS) classification. Psychological status was assessed using the depression score and the non-specific physical symptom score with subscales of the Revised Symptom Checklist-90 (SCL-90-R). The prevalence of reported pain in the face, jaws and/or TMJs was 14 and 9 per cent for the CLP and non-cleft group, respectively, and did not differ significantly between the groups. The CLP group exhibited a significantly reduced jaw-opening pattern (P < 0.001) and a higher frequency of crossbites (P < 0.05) compared with the non-cleft group. Whilst jaw function was similar in both groups, a few items, e.g. speech and drinking, were significantly more impaired (P < 0.01) in the CLP group than in the non-cleft group. There were no significant differences between the two groups concerning tension-type headache or psycho-social distress. The study found that overall TMD pain or psycho-social distress was not more common in this CLP group than in a non-cleft group. PMID- 11398557 TI - A longitudinal evaluation of craniofacial growth in a patient with Kabuki make-up syndrome: a case report. AB - The purpose of this investigation was to evaluate the craniofacial growth of a patient diagnosed with Kabuki make-up syndrome (KMS). Craniofacial growth was assessed by analysing lateral cephalometric radiographs with an interval of 12-15 months. They were taken from 6 years 9 months to 14 years 2 months. Angular and linear measurement analyses of the craniofacial complex showed a hypoplastic maxilla and a constricted maxillary basal arch width. The mandibular size was relatively large and had started to increase from 13 years 4 months. This resulted in a prognathic face caused by forward growth of the mandible and insufficient growth of the maxilla. The skeletal pattern was Class III. Open bite morphology with a steep mandibular plane (SN-MP), a relatively short ramus, and a large gonial angle were also observed. In this subject, the facial dysmorphism found in the maxilla and mandible may have been influenced by several factors. Connective tissue disorder, macroglossia, lower tongue posture, and tongue thrust swallowing have been identified as possible aetiological factors that may determine dysmorphism in the craniofacial complex in this KMS patient. PMID- 11398558 TI - Medicare advocacy and entertainment at IMS 2001 Family of Medicine meeting. PMID- 11398559 TI - State health officials hope for the best but prepare for the worst, should it ever come. PMID- 11398560 TI - Is communicating with patients via e-mail e-risky? PMID- 11398561 TI - Taking gender into account. PMID- 11398562 TI - The Algase Wandering Scale: initial psychometrics of a new caregiver reporting tool. AB - This paper reports on the Algase wandering scale (AWS), a 28-item questionnaire, based on five dimensions of wandering. With factor analysis, an eight-factor solution explained nearly 70 percent of the variance in ratings for 151 long-term care subjects and confirmed three of the structuring dimensions. Reliability of the AWS was examined for internal consistency and for inter-rater reliability. The AWS had an alpha of .86; subscale alphas ranged between .88 (persistent walking) and .57 (routinized walking). Inter-rater reliabilities, estimated through cross-rater comparisons of the AWS and subscales with a four-point judgement of wandering status, were moderately strong and no significant differences existed between two sets of raters. Validity of the AWS and its subscales was supported by examining their ability to differentiate wanderers and nonwanderers, by positive correlation with measures of cognitive impairment and with multiple parameters of observed wandering, and by negative or no correlations with nonwandering locomotion. Although the AWS may be a useful measure of wandering in long-term care settings, validation of its factor structure and evaluation in cross-cultural samples is needed. PMID- 11398563 TI - Characteristics of assaultive psychiatric inpatients: updated review of findings, 1995-2000. AB - The present paper reviewed the studies of the characteristics of assaultive psychiatric inpatients from 1994 to the present. These studies partially confirmed earlier findings that assaultive patients are older persons with a diagnosis of active psychosis or other neurological abnormality and histories of violence toward others and substance-use disorder. However, the recent studies also document the profile of a second group of assaultive patients: younger females with personality disorders and histories of violence toward others, substance-use disorder, and personal victimization. Since there is a paucity of published empirical research on Alzheimer's and dementia patients for this topic, the implications from the present findings for long-term care and clinical intervention are examined. PMID- 11398564 TI - Psychological features in persons at risk for familial Alzheimer's disease. AB - Persons at risk for inherited neurodegenerative diseases may experience symptoms of anxiety and depression because of concern over the possibility of developing the disease in the future. The purpose of this study was to assess psychological and emotional symptoms in persons at the age of risk for developing early-onset familial Alzheimer's disease (FAD). Their responses on a psychiatric rating scale (SCL-90-R) were compared with four groups: patients with mild FAD; head injury patients; patients with clinically diagnosed depression; and healthy control subjects. Mean scores of the at-risk FAD group were not statistically different than those of the controls. In contrast, the head injury and depressed groups had significantly elevated scores across the clinical scales. These results suggest that depression and anxiety are not prominent features in persons at genetic risk for early-onset familial Alzheimer's disease. Similar results have been found in studies of persons at risk for developing Huntington's disease, another autosomal dominant neurodegenerative disease. PMID- 11398565 TI - Impact of family visits on agitation in residents with dementia. AB - In an investigation of the impact of family visits on the behavior of 30 residents in dementia special care units, we found on direct observation that agitation decreased significantly during visits, but returned to the previous level within 30 minutes. No significant differences were found between visits by spouses or adult children. The premorbid quality of relationship was unrelated to family visitor enjoyment of visits or to differences between agitation level before and after visits. While 70 percent of visitors reported that they found visits pleasant (mean number of visits: 12.72 per month), 20 percent found visits unpleasant. Visitors' enjoyment or displeasure was significantly related to the difference between the number of pre-visit and post-visit agitated behaviors. Visitors indicated that the resident's mental status (33 percent) or difficulty communicating with the resident (30 percent) had the most negative impact on their visits. However, these factors appear amenable to education, suggesting that the quality of visits can be improved for visitor and resident with possibly greater positive impact on both. PMID- 11398566 TI - Male caregivers of spouses with Alzheimer's disease: risk factors and health status. AB - Caregiving for persons with Alzheimer's disease (AD) has been shown to pose a challenge to the health of the spousal caregiver. Because most of the caregiving literature focuses on the female caregiver, there is some question about the generalizability of such literature to the male caregiver. This report focuses on male caregivers of spouses with AD and represents a subsample from a larger descriptive study that examined the relationship between risk factors and the health status of spousal caregivers. Twenty-nine male caregivers affiliated with Alzheimer's organizations in Pennsylvania and Ontario, Canada, returned mail surveys. On average, physical health symptoms increased by one-third when comparing pre- and post-caregiving data. Caregivers also were experiencing moderate to severe depression and burden. Male caregivers generally rated their physical health as fair to excellent and exhibited fewer than expected physical health symptoms. Caregiver health was related to perceptions of stress surrounding the provision of activities of daily living (ADL) assistance, the frequency of behavioral problems, perceptions of stress associated with the AD spouse's dysfunctional behaviors, and satisfaction with leisure opportunities. The identification of the role that caregiver perceptions of stressfulness associated with caregiving and the need for leisure satisfaction offer important implications for community-based education and respite services to maintain health status for spousal caregivers. PMID- 11398567 TI - Long-term consequences of the Alzheimer's caregiver role: a qualitative analysis. AB - A qualitative analysis was performed on responses of 51 participants to open ended questions about the experience of being family caregivers for a loved one affected by Alzheimer's disease. Participants had been in the caregiver role for at least eight years. Results indicated a pattern of adapting successfully or unsuccessfully to the caregiver role, experiencing initial caregiver burden, finding relief in social support when available, and long-term distress or long term positive change. Memories and feelings were strong about individuals and institutions that had been helpful or indifferent many years earlier as the caregivers struggled to cope with their unplanned predicament. PMID- 11398568 TI - Obtaining family consent for participation in Alzheimer's research in a Cuban American population: strategies to overcome the barriers. AB - Cultural values and beliefs affect family attitudes toward participation in research. Significant resistance to allowing their elders with dementia to participate in clinical research was encountered in Cuban-American families. These families expressed concern about disturbing the elder's comfort (tranquilidad) and solitude (soledad). Furthermore, most believed that intervention would be futile. Feelings of guilt associated with nursing home placement may have been exacerbated by the suggestion that active intervention could be effective. Strategies to overcome these barriers included reduced emphasis on the potential superiority of the intervention to be tested, reassurance that contact with research staff was usually appreciated by participants, arrangements to talk with the family as a group about the study, and increased use of Spanish-language consent forms. PMID- 11398569 TI - The value of case management in the publicly funded substance abuse treatment system. The perspective of program directors, case managers and clients. AB - The many purposes of this article is to understand the role and value of case management from the perspective of program directors, case managers and clients. A survey of program directors from publicly funded substance abuse treatment programs in Boston was administered, and in-depth interviews with a sample of program directors, case managers, and clients were conducted. Case management allowed programs to serve more complex clients and increased time available for counselors to focus on the clinical needs of clients. From the perspective of case managers and clients, much of the value of case management came from educating clients about steps they could take to meet their needs and then supporting them in their efforts as they took these steps. Successful steps taken to deal with these needs helped lay the foundation necessary to confront the challenges of treatment. Program directors, case managers, and clients considered case management a valuable enhancement to substance abuse treatment. PMID- 11398570 TI - Community care management. Advanced practice nurses as care managers. AB - A group of advanced practice nurses partnered with a major insurer in the design and implementation of a care coordination model for high-risk older adults. This article will discuss the process of such an undertaking, highlighting the successes and barriers encountered. The key elements of this program included early identification and regular reassessment of each member's acuity level; fostering close partnerships between individual or teams of APRNs and groups of physicians; and uninterrupted clinical management of high-risk members across the health care continuum. This model was designed to achieve the following outcomes: to support the physician management of high-risk, chronic individuals; to increase or maintain the health of members; and to reduce health care costs. Outcome studies have demonstrated a substantial net savings by decreasing acute care admissions by 54%, reducing hospital days by 42%, and trimming primary care physicians' and specialists' visit costs by 37%. There was a 33% reduction in the overall costs of health care for members enrolled in this program. Physicians and members both rated their satisfaction with the APRN-based model of care as very high. PMID- 11398571 TI - Computer-assisted tracking of a case management program for the homeless. AB - Computer scan sheet technology was used to evaluate process and outcome variables in a case management shelter program for the homeless. Clients spent up to four months in this program, working on a number of goal areas, including housing, employment, drugs and alcohol, mental and physical health, and literacy. Using Tele-Form, a computer program that allows scan sheets to be designed on screen, case manager recording forms were developed that allowed interventions to be documented on a daily basis, while psychosocial goal areas were documented on scan sheets at intake and termination. Data from these scan sheets were fed into SPSS, a statistical program for the social sciences. Using the case management tracking guidelines developed by Frankel and LaPorte, 1998, the results of this study showed that scan sheet technology was an effective, efficient, and extremely cost-effective way to track case management. The analysis of the data and subsequent discussion suggested ways about how to make case management evaluation more uniform across the country. PMID- 11398572 TI - Health policy and case management. AB - The purpose of this article is to analyze the performance of and support for case management using a policy framework in order to increase case managers' awareness of policy making and facilitate successful planning for future policy initiatives. Feldstein's (1996) theory of opposing legislative outcomes indicates that legislation can be viewed on a continuum, ranging from legislation that meets the needs of the public to legislation considered to be in the self interest of the participants and legislators. The current health care system requires that case managers working for publicly funded health care organizations balance the need for stewardship of U.S. tax dollars and the health care needs of consumers. It is apparent from the literature that case managers are successfully achieving this balance. However, certain conditions should exist that allow for case manager decision-making that promotes effective and efficient utilization of health care resources. Case managers must work within the context of the health care policy environment. Realizing that it is more likely that the conflicts between stewardship and the provision of health care services will continue, case managers' knowledge and influence regarding policy making becomes imperative in order to ensure that these conflicting goals do not become mutually exclusive. PMID- 11398573 TI - Vision rehabilitation. A missing link in client care. PMID- 11398574 TI - Medicare home health coverage. PMID- 11398575 TI - Relieving stress. A short-term support group for home attendants. AB - Home attendants (HAs) work in relative isolation, burdened by conflicting demands. This article details an eight-session support group for HAs, designed to explore its effects on their work life. Meeting for hour-and-a-half sessions with no fixed agenda, the group offered members an opportunity to communicate with others in similar situations. Participants reported that the group experience helped relieve stress and made them feel less alone. Other benefits included gaining strategies for coping with difficult situations and learning to set boundaries. Further investigation into the effectiveness of similar groups is suggested. PMID- 11398576 TI - HIV/AIDS and older adults. The invisible ten percent. PMID- 11398577 TI - [Concentrations of MMP-1, TIMP-1, MMP-1/TIMP-1 and I CTP complexes in follicular fluid as related to fertilization rate in women treated with in-vitro fertilization]. AB - OBJECTIVES: Metalloproteinases and their inhibitors appear to control connective tissue remodelling during follicular rupture. The aim of the study was to establish if oocytes fertilisation rate after ovulation induction depends on the concentrations of MMP-1, its inhibitor TIMP-1, MMP-1/TIMP-1 complex and I CTP in follicular fluid (FF). MATERIAL AND METHODS: FF were collected from 37 infertile patients undergoing ovulation induction using either short or long protocol. FF was obtained 36 hours after administration of hCG (Pregnyl). The level of MMP-1, TIMP-1, MMP-1/TIMP-1 complex were measured using ELISA kits and I CTP, E2, FSH, LH, using RIA assay kits. RESULTS: Statistically significant (p < 0.05) difference was found in TIMP-1, E2, and FSH concentration, being higher in the group with more than 75% fertilisation rate: TIMP-1 728.8 + 100.1 ng/ml vs 666.3 + 94.5 ng/ml; E2 477.3 +/- 160.0 ng/ml vs 368.0 +/- 190.0 ng/ml and FSH 7.27 +/- 1.45 mIU/ml vs 6.24 +/- 1.6 mIU/ml. CONCLUSIONS: Statistically significant increase in TIMP-1 concentration observed among patients with fertilisation rate above 75% indicates an important role of this substance in ovulation process. PMID- 11398578 TI - [The effect of hormonal replacement therapy on symptoms of climacteric syndrome and selected metabolic parameters]. AB - GOAL OF STUDY: The goal of this paper was estimation of influence of HRT (after 3 and 6 months of treatment of 112 women) the symptoms of climacteric syndrome and depression associated with the said syndrome based on the scales of Kupperman, Green, Hamilton and Beck as well as some selected parameters of metabolic state- concentration of foloculotropic hormonal profile FSH and 17 beta-estradiol E2, lipid profile (TC, HDL, LDL, TG-A) and glucose concentration. CONCLUSIONS: 1. The implementation of hormonal replacement therapy on 112 women made it possible to conclude that the treatment caused withdrawal of symptoms of climacteric syndrome and depression associated with the said syndrome. 2. Hormonal Replacement Therapy to a great extent had a beneficial influence on the lipid profile with 112 women after menopause especially as far as LDL is concerned. 3. HRT caused enormously significant increase in 17 beta-estradiol concentration and decrease of FSH concentration. 4. HRT statistically had no impact on glucose level with the study group of women. PMID- 11398579 TI - [Integrins and prolidase activity in uterine leiomyoma during tumor growth]. AB - Collagens and proteoglycans are the main components of connective tissue. Prolidase plays an important role in the balance between collagen biosynthesis and degradation and its activity reflects the rate of collage turn-over. The activity of this enzyme is known to be regulated by interaction of collagen with beta 1 integrin receptor. It was found that increase in the uterine leiomyoma weight is accompanied by increased beta 1 integrin receptor expression and prolidase activity. It suggests that collagen turn-over is altered in uterine leiomyoma growth. PMID- 11398580 TI - [Usefulness of voiding history using voiding dysfunction questionnaire in diagnosis of lower urinary tract dysfunction in women]. AB - OBJECTIVES: The two functions of the lower urinary tract are the filling/storage and emptying. Similarly lower urinary tract symptoms can be subdivided. DESIGN: The aim of the study was the estimation of usefulness of present complaints voiding history in diagnosis of lower urinary tract dysfunctions. MATERIAL AND METHODS: 53 women with the mean age of 37.2 (range of 16 to 70) with chronic lower urinary tract symptoms without neurogenic or organic disease were examined. RESULTS: There was only a correlation between urge incontinence and bladder instability (p = 0.0075). CONCLUSION: There were no correlation (except urge incontinence) between symptoms and lower urinary tract dysfunctions. PMID- 11398581 TI - [Conservative treatment of patients with borderline ovarian tumors]. AB - OBJECTIVES: The aim of this study was to characterise group of patients with borderline tumours undergoing surgical treatment. DESIGN: The analysis included 83 patients with ovarian tumours of borderline malignancy at stage Ia, operated in the Gynaecological Department of Medical University of Gdansk between 1978 1997. The study takes into account comparison of: age of patients, type of surgery, tumour pathology, post surgical treatment. Furthermore, long term follow up was assessed. RESULTS: In the group of 83 patients with stage Ia 37 ware treated with conservative surgery, 46 underwent radical treatment. Postsurgical chemical treatment was not applied. Three patients had to be reoperated because of neoplasm disease recurrence. 8 patients were died from reasons not connected with main disease. CONCLUSIONS: Conservative surgery is proper treatment for young women with borderline ovary tumours in stage Ia. For older, perimenopausal women TAH with BSO without additional chemotherapy is suggested. PMID- 11398582 TI - [Clinical evaluation of Divina in treatment of hormonal disturbances in women]. AB - The aim of this study was the assessment of clinical efficacy during hormonal treatment with Divina among women with hormonal cycle disturbances. Divina is a estrogen-progesatagen drug containing 2 mg valerate estradiol and 10 mg medroxyprogesteron acetate. The influence on lipid profile, liver and kidney activity and glucose tolerance was assessed. The measurement of bone density was performed twice before and after 6 months of treatment with Divina. PMID- 11398583 TI - [Twin-to-twin transfusion syndrome--case report]. AB - Authors described a case of twin-to-twin transfusion syndrome that did not comply with a classic definition of syndrome. It has been presented possibilities of ultrasound screening regarding to Doppler blood flow velocimetry. It has been also shown the usefulness of these methods in the monitoring and assessment of fetal status. This is an example of Doppler blood flow velocimetry applying that yields information concerning fetal condition. This method seems to be very useful in these situations when fetal heart rate monitoring is difficult or even inapplicable. PMID- 11398584 TI - [Intrauterine therapy of obstructive uropathy--case report]. AB - The use of vesicoamniotic shunt in the case of obstructive uropathy was described. On the base of ultrasound screening it has been diagnosed urethral obstruction. This abnormality of urinary tract was found in male fetus. It has been diagnosed the posterior urethral valve. Qualification to intrauterine therapy was based on normal parameters of renal function. Under ultrasound guidance the vesicoamniotic shunt was installed. The normal drenage was observed within 6 weeks, until it was dislocated. In 36th week of gestation the cesarean section was performed. The indications were signs of urethral obstruction. The boy was born in whom posterior urethral valves were recognized. He was born in good condition without any biochemical features of renal damage as well as signs of hypoplastic lung. PMID- 11398585 TI - [Peritoneal hemorrhage after rupture of the uterine ligament latum venous plexus during the 26th week of pregnancy]. AB - The article presents a case of spontaneous rupture of the uterine venous plexus on the right side ih a 32 year old multipara in 26th week of pregnancy. Attention is called to diagnostic difficulties of this complication and methods of controlling venous bleeding are discussed. PMID- 11398586 TI - [Bacterial and fungal vaginal infections--immunologic and clinical aspects]. AB - Bacterial and fungal vaginal infections seem to be one of the most important problem in clinical gynaecology, because of their very often incidence and recurrence. One of the most important factor that implicates those epidemiological facts is local immunological response. Present paper summarises immunological mechanisms involved in antimicrobial response, and current knowledge on diagnosis and treatment of bacterial and fungal vaginal infections. PMID- 11398587 TI - [Coeliac disease and problems associated with reproduction]. AB - Coeliac patients taking a normal diet have a shortened reproductive period with delayed menarche and early menopause. Many studies have shown that coeliac women are susceptible to reproductive difficulties such as infertility and miscarriages. The disease is also associated with low birth weight in babies and short duration of breast feeding. Folic acid deficiency is present in the majority of patients with untreated disease and it might be a maternal risk factor for neural tube defects and orofacial clefts. Coeliac men may have reversible infertility, and as in women, if gastrointestinal symptoms are mild or absent the diagnosis may be missed. It is important to make diagnosis because the giving of gluten free diet may result in conception and favourable outcome of pregnancy. PMID- 11398588 TI - [Quality of life and working status in patients with coronary artery disease treated by percutaneous coronary angioplasty during long-term observation]. AB - Improvement in the quality of life is nowadays widely acknowledged as an important factor in assessment of the long-term outcome of the actually pursued treatment. Percutaneous transluminal coronary angioplasty (PTCA) has been used in the treatment of coronary disease in ca. 1/3 of patients. The present study aimed to assess the quality of life in 567 patients successfully treated by PTCA in the period 1987-1996. In the observation period spanning 2.5-12 years (5.5 years on average) 26 patients (4.6%) died. A specially designed questionnaire was sent out to the remaining 541 patients, in which they were asked to attempt a subjective assessment of their general health condition and physical fitness, as well as to comment on the quality of their family and social life following angioplasty. The filled-in questionnaire was returned by 447 patients (82.6%). Their functional status, assessed in compliance with the CCS criteria, was perceptibly improved after the procedure and at the end of the observation period 70.5% of patients had no significant coronary complaints. A durable improvement after PTCA was claimed by 85.7% of patients, for 38.2% the result of PTCA turned out to be much better than originally expected, while 47.9% admitted that the procedure actually complied with their expectations. No improvement whatsoever was reported by 11.8%, while 2.3% of patients claimed their condition to have deteriorated since. In their own assessment, normal sexual life after PTCA was still pursued by 63.1% of patients, though men appeared to be more active in that domain than women (68.0% vs. 44.7%, p = 0.0001). Notable improvement was reported by 15.2%, while deterioration was claimed by 17.2%; in both cases it was significantly more frequent in men (improvement 17.0% vs 8.5%, p = 0.042, deterioration 19.5% vs 8.5%, p = 0.012). Angioplasty had no impact on their social relations in 78.8% of patients, while 21.2% of them are commonly perceived as the disabled persons. The need for psychotherapy (counselling) was expressed by 15.9%. The analysis of the key factors determining the actual quality of life in patients, carried out in the sub-groups, did not yield significant differences between the patients with stable and unstable angina, patients below and over 50 years of age, patients with complete and incomplete revascularisation, nor in patients who experienced or not, cardiac events during the follow-up. The significant differences between men and women were encountered only with respect to their sexual life. Diabetics in comparison with non-diabetics significantly more frequently reported improvement in their sexual life, full resumption of their non-occupational activities, generally showed more appreciation for the overall outcome of the PTCA procedure. Diabetics much less frequently experienced improvement in their general physical fitness, generally regarded the need for regular medical check ups as a hindrance, as well as were more frequently perceived as the disabled persons. The working status of the patients after angioplasty failed to improve, however, as nearly half of them, despite the successfully completed procedure, was granted disability pensions or took advantage of the voluntary retirement schemes. Only 7.2% of the non-working patients well within their statutory employment age successfully returned to their employment, although not on a full time basis. Patients who had completed their primary or secondary education were found to leave their employment twice more frequently than the ones with academic education. Similarly, patients living in the rural areas or in small towns (less than 40.000 inhabitants) left their employment twice more frequently than the ones living in major cities. CONCLUSIONS: Both the functional status of patients and the majority of the analysed factors determining the quality of life--in the subjective assessment of those patients--improved significantly after a successful PTCA. This improvement is irrespective of the patients' age, symptoms of unstable angina during the procedure, extensiveness of revascularisation as well as cardiac events during the follow-up. Nearly half of the working patients tend to leave their employment after a successful percutaneous revascularisation, while the non-working patients are seldom prompted to resume their employment by the successful procedure. Patients with academic education, living in major cities, are usually found to resume their original employment most frequently. PMID- 11398589 TI - [Post-traumatic bacterial meningitis]. AB - 10 patients with posttraumatic bacterial meningitis were treated in the Department of Infectious Diseases of the Jagielloniam University--Collegium Medicum during the period of 63 months. Traffic accidents were responsible for 80% of all cases of trauma. The most common place of injury was the base of anterior cranial fossa. Infecting agent was established in 8 cases. Gram-positive aerobic cocci (Streptococcus pneumoniae, Enterococcus faecalis, and Enterococcus spp.) were found in 4 patients, Gram-negative aerobic cocci (Neisseria meningitidis) in 2 patients, and Gram-negative aerobic rods (Acinetobacter baumanii, Pseudomonas aeruginosa, Klebsiella spp, Escherichia coli) in 3 patients. Streptococcus pneumoniae was still highly sensitive to penicillin, ampicillin and cefotaxime. Enterococcus faecalis and Enterococcus spp. were sensitive to vancomycin and teicoplanin. Neisseria meningitidis was sensitive to penicillin, cefotaxime, amoxicillin/clavulonate. Gram negative aerobic rods were sensitive to third-generation cephalosporins, carbapenems and aminoglycosides. Among 10 patients treated for posttraumatic bacterial meningitis 1 patient died and in 1 preserved vegetative state was diagnosed. In 4 patients severe or moderate disabilities developed, however 4 recovered completely. 6 patients after regression of the CNS infection were qualified to neurosurgical intervention. In 3 patients reconstructive operation of the basis of anterior cranial fossa with tissue glue Beriplast P. was done, the recovery was complete. 3 patients didn't agree to surgical operation, and we don't know what has happened with them. They have not come to the control visit. PMID- 11398590 TI - [Use of modern microscopic techniques for examining dialysis membrane properties]. AB - The aim of the study was the microscopic evaluation of internal structure of cuprophane and polysulfone membrane and their surface analysis before and after reprocessing. The investigations were performed using an optical measurement system (Digital Instruments), a scanning electron microscope (SEM) and an atomic force microscope (AFM). We confirmed by SEM that reprocessing completely removed biofilm from both membranes surface. The analysis based on AFM visualized channels in the examined membrane. The diameter of the channels varied from 150 nm for cuprophane to 380 nm for polysulfone. The roughness expressed as root mean square (RMS) was higher for cuprophane than for polysulfone membrane. The physical differences between nanostrucure of the examined membranes might be responsible for lower biocompatibility of cuprophane. PMID- 11398591 TI - [Measurement of fluid compartments using electrical bioimpedance for assessment of target weight in hemodialysis patients]. AB - The prescription of optimal hydration status in hemodialysis patients remains a much disputed topic in dialysis treatment. In particular, assessment of the patients optimal weight ("target weight") poses considerable difficulties. Multifrequency bioimpedance spectroscopy analysis (BIS) has been recommended as a non invasive, practical, and relatively non expensive method to determine hydration and nutritional status in patients on maintenance hemodialysis (HD). In the current study we used whole body BIS analysis for determination of body water (BW) compartments; total body water (TBW), extracellular water (ECW), and intracellular water (ICW) in 133 healthy adults, and in 227 hemodialysis patients with end-stage renal disease. BIS results were compared to anthropometric measurements. Our results showed strong correlation between TBW measured by BIS in control group in comparison to anthropometric calculation (p = 0.001). In HD patients we observed higher range of TBW, and TBW/ECW ratio (from 15.6 to 56.1 L and from 0.33 to 0.78), as measured by BIS at pre-HD, and also post-HD period (TBW ranged from 13.1 to 56.2 L, ECW/TBW ratio ranged from 0.33 to 1.27). The TBW BIS results did not correlate with anthropometric calculation. We postulate using of multi-frequency bioimpedance technique in precise determination of fluid compartments and in consequence in the assessment of "target weight" in hemodialysis population. PMID- 11398592 TI - [Selected parameters of bone metabolism in hemodialyzed and peritoneally dialyzed patients]. AB - Renal osteodystrophy is a metabolic bone disease occurring in patients with end stage renal failure. The aim of the study was to compare serum concentrations of some bone markers in hemodialysed (HD) patients and in patients undergoing continuous ambulatory peritoneal dialysis (CADO). We studied two groups of patients with end-stage renal failure: 52 hemodialysed individuals aged 24-74 years and 19 peritoneally dialysed patients aged 20-70 years. Serum calcium and phosphate concentration, cholesterol, triglycerides, total protein, albumin, alkaline phosphatase, urea before and after HD, urea in CADO patients were determined by standard laboratory methods. Serum PTH, osteocalcin, 1,25(OH)2D3 and insulin-like growth factor (IGF-1) concentrations were measured by commercially available radioimmunoassay. Serum tumor necrosis factor (TNF-alpha) and interleukin-1 (IL-1) concentrations were measured by ELISA. There were no differences between serum concentrations of the studied bone markers in hemodialysed patients and CAPD patients. All dialysed patients presented high concentrations of serum PTH, osteocalcin, alkaline phosphatase activity, lower serum IGF-1 concentration and normal serum calcitriol concentration. High serum PTH and osteocalcin concentrations may indicate intensification of bone synthesis, what is typical for osteitis fibrosa. PMID- 11398593 TI - [Levels of ionized magnesium in women with various stages of postmenopausal osteoporosis progression evaluated on the basis of densitometric examinations]. AB - The aim of the work was the estimation of the ionized magnesium level in postmenopausal osteoporotic women in different stage of osteoporosis progression assessed in densitometrical investigation. The study included 53 women, divided due to T index into 3 subgroups compatible with WHO osteoporosis classification criteria. The control group included 20 healthy women at perimenopausal age. In the group of women with osteoporosis and severe osteoporosis significantly lower ionized magnesium level was determined in comparison with the control group and the group of women with osteopenia (p < 0.05). No correlation between T index (assessing the stage of osteoporosis progression) and hypomagnesaemia was found. PMID- 11398594 TI - [Levels of alpha-2-macroglobulin in blood serum of women giving birth to hypotrophic and eutrophic newborns]. AB - We tested the concentration of alpha-2-macroglobulin in sera of 33 women bearing eutrophic newborns, 36 women bearing hypotrophic newborns and 30 healthy not pregnant women in reproductive age. The concentration of this inhibitor was measured using radial immunodiffusion method according to Mancini et al. We found distinct decrease of alpha-2-macroglobulin concentration in sera of bearing women. In women bearing eutrophic newborns we found 154 mg/dl, in women bearing hypo-trophic newborns 171 mg/dl whereas in controls 250 mg/dl. We have noted statistically significant differences between tested groups to controls and between investigated groups. Taking under consideration the role of alpha-2 macroglobulin as the modulator of immune system as well as the activity of several cytokins, therefore one can suppose that alpha-2-M may affect on cellular growth developed foetus in intrauterine. PMID- 11398595 TI - [Is surgical treatment of hyperthyroidism in pregnancy reasonable?]. AB - Hyperthyroidism is second to diabetes mellitus as the most common endocrinopathy in pregnancy. It is usually caused by Graves' disease. The appropriate treatment is the only way to prevent the incidence of dangerous complications both to mother and foetus, including thyrotoxic crisis. Most cases of hyperthyroidism during pregnancy can be successfully treated with thyrostatics (propylotiouracil, methylotiouracil in the lowest efficient doses). However, the surgical treatment (sub-total thyroidectomy within the second trimester of gestation, soon after the euthyroid state following short medication is reached) is still an elective approach in selected cases: 1) allergy to antithyroid drugs, 2) large compressive goitre, 3) suspicion of thyroid cancer, 4) patients who require large doses of antithyroid drugs to reach and sustain euthyroid state, 5) poor patient compliance, 6) extremely rare resistance to antithyroid drugs. The study included 24 cases of pregnant women, who underwent surgery due to hyperthyroidism. The analysis of indications to surgery and postoperative pregnancy, birth and puerperium course was performed. All the patients gave birth on time to healthy children. No negative influence of neither preoperative antithyroid treatment nor surgical procedure on pregnancy delivery, further psychomotoric children's development and health was found during 36 months (+/- 20 months) of postoperative follow up. It indicates on usefulness, efficacy and safety of surgical treatment in selected cases of hyperthyroidism during pregnancy. PMID- 11398596 TI - [Causes of premature labor in pregnancy complicated by diabetes depend on diabetes progression]. AB - The aim of this study was to demonstrate what factors influence preterm termination of pregnancy complicated by diabetes, in dependence of diabetes progression. The following factors have been analysed: rate and causes of preterm labours according to White classification, mean duration of pregnancy, mode of delivery, birth weight, Apgar score and diabetes control in the preterm delivered women with different progression of disease. As a result we observed, that in G/A and G/B classes and in short lasting class B, the most common cause of preterm labour was spontaneous delivery and prematurity rate was low, oscillated between 1.7 and 15.8 percent. In those classes we also observed that diabetes control was very good or adequate. In D, R, F, and RF classes the most common cause of preterm labour is intrauterine fetal distress, that causes preterm termination of pregnancy, despite immaturity, and prematurity rate is 30-50 percent. We observed bad diabetes control in above mentioned classes. PMID- 11398597 TI - [Etiopathogenesis of diabetic neuropathy]. AB - The cause of diabetic neuropathy remains unclear but several mechanisms have been considered. Hyperglycemia with all its metabolic and vascular consequences probably plays the main role. Metabolic consequences of hyperglicemia including increased level of sorbitol and fructose, myoinositol deficiency and Na+/K+ adenosine triphosphate deficiency alter the function of peripheral nerves and lead to an oxidative stress, which changes fatty acid metabolism. Chronic hyperglycemia is also responsible for abnormalities in microcirculation that could lead to local ischemia evoking many endothelial changes. There are also some genetic factors, impaired neurotrophic support and autoimmune damage that influence sensitivity to peripheral nerve damage in diabetes mellitus. PMID- 11398598 TI - [Chronic complication of diabetes and homocysteine]. AB - Increased homocysteine levels are associated with increased risk of cardiovascular disease incidence and mortality in general population and with even greater risk in persons with diabetes mellitus. Among possible mechanisms of proatherogenic action of homocysteine increased procoagulant activity, increased oxidative stress and effect on proliferation of smooth muscle cells should be mentioned. However, the role of hyperhomocysteinemia in the development of microvascular complications is the subject of a few studies with controversial results. Some studies indicate the role of hyperhomocysteinemia in predicting micro- and macroalbuminuria and also retinopathy and neuropathy. Serum homocystine levels are dependent on age, renal function, and dietary intake of folic acid and B vitamins. The role of insulin in homocysteine metabolism is the subject of studies. Determination of the role of hyperhomocysteinemia in macrovascular and microvascular diabetes complications could be of importance in their prevention through dietary and pharmacological modifications of homocysteine levels. PMID- 11398599 TI - [Immune markers in preclinical phases of diabetes type 1. Role of cytokines, adhesion molecules and peripheral blood lymphocyte subpopulations in the pathogenesis and prediction of insulin-dependent diabetes]. AB - Multidirectional studies estimating the alterations of cellular immune system in the preclinical phase of type 1 diabetes mellitus create the possibilities for better understanding the mechanisms of the auto-aggression process leading to the pancreatic beta cells destruction and insulin-dependent diabetes mellitus development. In the present review we analyse the recent studies concerning the alterations of the cellular immune markers (ex. peripheral blood lymphocytes subsets, cytokines production or adhesion molecules expression, etc...), which could reflect an early stages of autoimmune process and better identify people at risk of type 1 diabetes mellitus. In summary we suggest that the measurements of new immune markers of prediabetes, together with the estimation of the pancreatic auto-antibodies, genetic risk and age of the studied subjects give the possibility for the better early diagnosis of the people at high risk of type 1 diabetes mellitus development. There is an increasing hope that these markers are useful for the estimation of the efficacy of the preventive therapeutic procedures aiming to type 1 diabetes mellitus protection. PMID- 11398600 TI - [The effect of alcohol on bone mineral density in men]. AB - Alcohol abuse is an essential factor promoting an excessive reduction of bone mass during the normal male aging. Authors present an issue of osteopenia and osteoporosis in men, that sometimes is due to the long-term bone tissue exposition on ethanol. They also present actual views on the pathogenesis of this pathology. Ethanol impairs mainly an osteoblastic activity that results in reduced bone formation and mineralisation. Affecting the osteoclasts' function, alcohol is also able to induce bone resorption. Despite the direct toxic effect of ethanol on bone tissue, the indirect influence of ethanol on metabolism of hormones participating in bone homeostasis has been revealed. In chronic alcoholics the deficiency of active metabolites of vitamin D is often observed. The findings on parathormone activity in alcohol male abusers are not consistent. Additionally, chronic alcoholism is frequently accompanied by an increase of cortisol levels, andropenia, dietary, vitamin and electrolyte deficiencies. PMID- 11398601 TI - [The significance of estrogens on metabolism of bone in men]. AB - Authors present actual data on estradiol, its production and systemic functions, with special stress on relations between the hormone (and estrogen-like substances) and male bone metabolism. Based on literature studies and cases of men with impaired hormone metabolism (estrogen resistency and aromatase deficiency), attention is paid to the role of estrogens in the patophysiology of osteopenia and osteoporosis. PMID- 11398602 TI - [Neurohumoral mechanisms in pathophysiology of chronic heart failure]. AB - Pathogenic mechanisms of chronic systolic heart failure are constantly of great interest. In recent years the neurohumoral theory of heart failure has gained attraction. According to this theory, neurohumoral mechanisms play the main role in the pathogenesis of heart failure, especially in its progression. These mechanisms can be divided into 2 categories: vasoconstrictive, mitogenic and antinatriuretic on the one hand and vasodilative, antimitogenic and natriuretic on the other one. The former consists of sympathetic nervous system, renin angiotensin-aldosterone system, vasopressin, endothelin, cytokines. The latter comprises natriuretic peptides, prostaglandins and nitric oxide. Undoubtedly unfavourable roles of sympathetic system and renin-angiotensin-aldosteron have been shown in the progression of heart failure. Data are being also gathered confirming harmful effects of endothelin and cytokines and possibly of neuropeptide Y and vasopressine. Extensive data exist that demonstrate beneficial influence of natriuretic peptide on heart failure. The roles of nitric oxide as well as recently discovered adrenomedullin and medullipin are far from clear. PMID- 11398603 TI - [Hepatocyte growth factor in kidney diseases]. AB - HGF--hepatocyte growth factor--belongs to the growth factors family, derivatives of plasminogen. Authors in this study discuss HGF and its receptor structure, physicochemical properties, release of HGF and its action on cellular level, describe also methods of HGF detection. HGF is a potent mitogen for hepatocytes, plays an important role in liver regeneration after inflammation and partial hepatectomy. Elevated levels of serum HGF were detected in liver diseases with co existing nephromegaly. HGF has also renotropic properties. A role of HGF in kidney development in physiology and pathology conditions, HGF action in acute renal failure, in chronic renal failure and after kidney transplantation was discussed. According to current literature, HGF importance in glomerulonephritis was described. Elevated concentration of this growth factor correlates with kidney destruction and could be connected with glomerulopathy exacerbation. HGF also plays a role in vascular endothelium defence and regeneration. The importance of HGF in arterial hypertension pathomechanism was underlined. PMID- 11398604 TI - [Basic terminology for describing hemodialysis fistula]. AB - Many specialists: nephrologists, surgeons, radiologists and nurses pay much attention to dialyzed patient using different terminology. The presented work establishes the principal rules of describing hemodialysis fistula to find convergence in terminology. PMID- 11398605 TI - [Long-term good results of surgical treatment for spontaneous epi- and subdural hematoma in a female patient on maintenance hemodialysis]. AB - Spontaneous intracranial hematoma is not rare, but with bad prognosis, complication in patients on maintenance hemodialysis (HD). Diagnostic difficulties result from a fact that symptoms of acute hematoma such as headaches,, nausea, vomitis, apathy, sleepiness, parestesia and seizures may also suggest dysequilibrium syndrome, dialytic dementia as well as hypertensive encephalopathy. We describe a case of female patient with 20-year interview data of hypertension on HD since 1981 because of end-stage renal failure in a course of chronic glomerulonephritis, who developed spontaneous epi- and subdural hematoma four year ago in 47 age of life. Performed CT examination confirmed diagnosis and on the same day the patient underwent right frontoparietotemporal craniotomy and the hematoma was removed. During postoperative period, HD sessions were performed without heparin. After surgery the patient developed transcient hypertonia, epileptic sizures and left-sided paresis. Currently, 48 months after craniotomy the patient is fully rehabilitated, with normal blood pressure, without epileptic sizures or palsy. Gradually we discontinued anticonvulsans and antihypertensives. PMID- 11398606 TI - [A case of isolated gallbladder rupture after blunt abdominal trauma]. AB - Isolated injury of gall bladder as a result of blunt abdominal trauma is very rare. We report a case of 39 year old man, who was admitted to our Clinic during emergency. He presented upper right quadrant abdominal pain, increasing on palpation and guarding. Ultrasound examination showed fluid under liver. Disruption of the neck of the gall bladder was found during laparotomy. Cholecystectomy was performed. Patient was discharged from hospital five days after the operation. PMID- 11398607 TI - [First Polish radiologic examinations]. AB - Authors present information about first Polish experiments with X-Rays made on January and February of 1896. PMID- 11398608 TI - Split-course radiotherapy: where do we stand? AB - BACKGROUND: Split-course radiotherapy is only rarely applied in curative radiotherapy and there might be a number of arguments to believe that continuous radiotherapy is superior to split-course treatment. In order to point out the evidence current treatment practice is based on, the available randomized trials and some prominent retrospective analyses on split-course radiotherapy were critically assessed. MATERIAL AND METHODS: The analysis of the clinical results was based on published data only. Publications were searched in a Medline database. RESULTS: Assessment of 13 randomized trials, including the data of 2,112 patients, revealed no significant difference between continuous-course and split-course radiotherapy. Astonishingly, the outcome of 77 radiotherapy studies on split-course, most of which are retrospective, seems to depend on the year of publication, suggesting publication bias. CONCLUSIONS: No clinically relevant difference between continuous and split-course radiotherapy could be found. This, of course, does not proof that there are indeed no differences but the data do not allow to draw clear-cut conclusions in favor of or against split-course radiotherapy due to methodological shortcomings of the studies. PMID- 11398609 TI - [Radiotherapy for basal cell carcinoma. Local control and cosmetic outcome]. AB - BACKGROUND: The basal cell carcinoma which is often occurring in the elderly can be well treated by surgery. For large and recurrent lesions and in cosmetically difficult locations external beam radiotherapy provides an equally effective treatment alternative. PATIENTS AND METHODS: From 1986 to 1999, 60 females and 39 males received primary radiotherapy for a total of 127 histologically verified basal cell carcinoma lesions. Tumors were mostly localized in the face at the temple, nose and forehead. Radiotherapy was applied with orthovoltage equipment and energies of up to 100 kV. Single doses ranged from 2 to 5 Gy related to the 80%-isodose depth. Weekly doses ranged from 8 to 25 Gy and total doses from 25 to 60 Gy. The mean follow-up period was 36 +/- 21 months. The acute sequelae were scored according to CTC criteria. Radiogenic late effects as single events were related to the radiation portal. RESULTS: 3 months after treatment all besides one patient (99%) experienced complete tumor remission (CR). In all cases, acute radiation reaction occurred within the radiation portal: CTC Grade 1 in 100%, CTC Grade 2 in 54% and CTC Grade 3 in 30% of the cases. All side effects regressed under simple local measures without further complications. Late sequelae were observed in three cases. Overall cosmetic outcome was good to excellent in almost all patients (98%). In two cases (2%) a local recurrence was observed 6 and 20 months after radiotherapy. CONCLUSION: External beam (orthovoltage) radiotherapy is very effective and yields high tumor control rates and good cosmetic results in long-term follow-up. Former dermatological treatment concepts should be replaced by an ICRU-based radiophysical dose prescription and should respect the newer radiobiological fractionation principles. PMID- 11398610 TI - [Radiotherapy for an adenolymphoma of the parotid gland (Warthin tumor)]. AB - BACKGROUND: With 17.6% of all primary parotid neoformations the benign Warthin's tumor (cystadenolymphoma) is the second common parotid gland tumor. Males > 50 years are affected predominantly. After surgery the recurrence rate is less than 5%. Histomorphologically the tumor is characterized by cystoid ducts lined by epithelial cells as well as lymphoid stroma. The lymphoid component has been described as radioresponsive whereas the epithelial parts are less radiosensitive. Since 1960 only one patient treated by primary radiotherapy has been published. CASE REPORT: A 77-year-old woman suffered from cystadenolymphoma (maximal diameter 7 cm). Because of its extension and the reduced performance status of the patient surgery was no option. Radiotherapy was performed with a total dose of 50 Gy. Clinically, the tumor regressed completely after 30 Gy, which was confirmed by CT at 6 weeks after completion of radiotherapy. After 6 and 12 months the patient stayed free of tumor. EPICRISIS: In our case the cystadenolymphoma was unusually large (7 cm). Radiotherapy with 50 Gy induced complete tumor regression. The good clinical response after 30 Gy suggests that the necessary dose may be lower for less extended cystadenolymphomas. CONCLUSION: We present a case of cystadenolymphoma treated by radiotherapy with 50 Gy resulting in a complete remission. Due to missing published experiences no common recommendation for the total dose can be given. In the following situations radiotherapy should be considered: 1. high surgical risk of damage to the facial nerve, 2. unfavorable cosmetic outcome after surgery, 3. inoperability for internal risks, 4. refusal of operation. PMID- 11398611 TI - Intracavitary afterloading boost in anal canal carcinoma. Results, function and quality of life. AB - BACKGROUND: First clinical data on a new intracavitary afterloading boost method for anal canal carcinoma is reported. PATIENTS AND METHODS: 20 consecutive patients (T1 5%, T2 70%, T3 20%, T4 5%; N0 75%, N1 10%, N2 15%; all M0) treated with external beam pelvic radiotherapy (median dose 56 Gy, range 46-64 Gy), simultaneous 5-FU and mitomycin (in 75%) and an intracavitary afterloading boost (one or two fractions of 5 Gy at 5 mm depth) were analyzed after a mean +/- SD follow-up for living patients of 4.4 +/- 2.1 years. Quality of Life (QoL) and anorectal manometry parameters were assessed in ten colostomy-free survivors. RESULTS: Overall, recurrence-free and colostomy-free survival at 5 years were 84%, 79% and 69%, respectively. No death was tumor-related. The only local failure was successfully salvaged by local excision. All three colostomies were performed for toxicity. Resting pressure and maximum squeeze pressure of the anal sphincter were reduced by 51% and 71%, as compared with control subjects, but quality of life was similar compared to healthy volunteers. CONCLUSION: The described regimen is highly effective but associated with increased toxicity. PMID- 11398612 TI - [Experiences with a clinical cancer registry for radiotherapy]. AB - BACKGROUND: A clinical cancer registry is required to assure result quality in radiation oncology. A simpler system could help small radiotherapy departments to survey their therapy results, as long as there is no connection to a central institution established. METHODS: Our clinical cancer registry RADOC was designed for easy handling on a single working place with standard Windows PC. RESULTS: Without additional costs, we store about 550 treatment courses each year and follow-up data. Several times a year the program provides basic analysis of these data for our internal quality assurance. As an example, the results on breast cancer are shown. CONCLUSIONS: Our department's internal therapy registry enables us to survey continuously important therapy results without additional costs, and thereby adds to our quality assurance process. On the other hand, only a centralized registry can provide complete data. PMID- 11398613 TI - Patient position reproducibility in fractionated stereotactically guided conformal radiotherapy using the BrainLab mask system. AB - PURPOSE: Dedicated mask systems nowadays allow the use of stereotactic radiotherapy in fractionated regimes, therefore combining the advantages of high precision radiotherapy with the biological benefit of fractionation. Therefore the knowledge of institution specific isocenter accuracy is essential for decision-making about margins to be allowed to form the planning target volume. PATIENTS AND METHOD: Measurements of isocenter deviations during fractionated treatments were performed in 33 patients using the simulator Simulix-xy (Oldelft) in connection with the BrainLab angiographic localizer-box as well as port-films. In both cases repeated images were overlaid by use of anatomical landmarks with a methodical accuracy in the order of 0.5 mm. RESULTS: Both methods yield random isocenter deviations of less then 2 mm (standard deviation) in all three directions and no significant systematic deviations. These values are in the order of the accuracy of the method, obtained by comparison of two independent investigators, as well as they are comparable with the literature. CONCLUSIONS: The accuracy of less than 2 mm indicates safety margins of 3-4 mm as sufficient for clinical routine to cover the target in 95.5% of all set-ups (2 SD). PMID- 11398615 TI - [Porgnostic impact of tumor localization in breast cancer]. PMID- 11398614 TI - [Percutaneous radiotherapy plus brachytherapy and concurrent chemotherapy for patients with localized esophageal carcinoma--Radiation Oncology Group Study 9207]. PMID- 11398616 TI - [Combination therapy with buserelin and tamoxifen in metastatic breast cancer in premenopause: a randomized study]. PMID- 11398617 TI - [Aromatase inhibitors: anastrozole versus tamoxifen as first-ine therapy for advanced postmenopausal breast cancer]. PMID- 11398618 TI - [Computer-assisted, robot-assisted hip prosthesis--standard procedure or specialty indication?]. AB - Approximately 150,000 total hip replacements annually are performed in Germany, with an increasing incidence of implants carried out on young subjects. Due to aseptic loosening, this group will undergo some revision arthroplasty after 15 years. Therefore, a permanent fixation appears mandatory. In case of posttraumatic or postoperative anatomical changes, exact preoperative planning by x-rays is extremely difficult. This might result in a higher rate of complications (e.g., displacement of the implanted prosthesis, fractures of the proximal femur), as documented by the available literature. The preoperative planning with CT images obtained at the 3D workstation shows the exact cortical situation, while the robot-assisted surgery allows the precise execution of the preoperative plan during surgery. By this point of time, long-term results of computer-guided, robot-assisted implantation of endoprosthesic devices are still lacking. However, the preliminary data indicate good results by means of anatomical position of the endoprosthesis and perioperative complications. PMID- 11398619 TI - [Knee joint endoprosthesis]. AB - Total knee replacement is increasing in number due to increased life expectancy and improved implants. [figure: see text] Degenerative changes (arthritis) and joint destruction based on polyarthritis and posttraumatic deformities are the common indications. There are basically four modes for knee replacement: Unicompartimental prosthesis, non-constrained, semi-constrained (posterior stabilized) and constrained total knee arthroplasty. Of major importance in knee alloarthoplasty are restitution of the correct limb axis, sufficient lateral ligaments and the restoration of the patellofemoral joint. Therefore particularly in cases with deformities an extensive soft tissue release is mandatory. A meticulous operative technique and versatile implants lead to good clinical outcome. Possible complications are still septic and aseptic implant loosening as well as wound infections. PMID- 11398620 TI - [Shoulder joint endoprosthesis in the treatment of multiple fractures of the humeral head. Development and value]. AB - Fractures of the humeral head are still challenging to decisions of indications, surgical skills and physiotherapy. 3- and 4-part-fractures of the humeral head in younger adults are still dominated by reconstructive osteosynthesis. New developments in osteosynthesis materials allow the surgeon to make use of minimal invasive techniques more often. Treatment of elderly patients with these injuries has changed within the last years. When it is obvious that the goal of a stable osteosynthesis can not be reached, shoulder prosthesis has become a primary treatment in elderly patients. New designs in shoulder prosthesis seem to lead to better functional results. PMID- 11398621 TI - [Total ankle joint replacement]. AB - We evaluated the short- to mid-term results of an unconstrained total ankle prosthesis (S.T.A.R.) with uncemented fixation. Seventy-nine consecutive ankle replacements were performed in 76 patients between 1996 and 2000. The initial diagnosis was posttraumatic osteoarthrosis in 51 cases (64%), primary osteoarthrosis in 14 cases (18%), and systemic joint affection in 14 cases (18%). Although 29 revision operations were necessary, all implants became fix, and no implants have been removed. At the last follow-up control, 38 patients (62%) were very satisfied, 20 patients (33%) were satisfied, and 3 patients (5%) were satisfied with reservations. The obtained range of motion was 35 degrees (range, 15 to 55 degrees), with a maximal plantarflexion of 27 degrees (range, 15 to 45 degrees) and dorsiflexion of 8 degrees (-3 to 20 degrees). The AOFAS-Hindfoot Score of the 61 patients scored 84 points (range, 51 to 100 points). After settling of the implants within 6 weeks, no migration was noted in any case, and all implants were considered to be stable. The favorable results were considered to be a result of the mechanical properties of the S.T.A.R. total ankle prosthesis that allow for unconstrained motion. The rate of revisions, however, was too high, and it was considered to be a result of the design of tibial and talar component, the tibial fixation and the poor instrumentation. Nevertheless, total ankle replacement was shown to be a viable alternative to ankle arthrodesis. A longer follow-up is, however, mandatory to evaluate the long-term prognosis. PMID- 11398622 TI - [The infected endoprosthesis with the example of the hip joint endoprosthesis. An increasing danger to patient and society]. AB - Because of demographic factors there is an increase in the numbers of total joint replacement operations each year. Deep infection after joint replacement remains one of the major complications in orthopedic surgery. The economic consequences for society are enormous. The treatment of such infections usually means a long and difficult course for the patient. In most cases multiple operations, including removal or exchange of the prosthesis are required. Concepts of treatment are very different and vary from debridement and keeping the prosthesis, resection arthroplasty to one- and second-stage exchange procedures. It is essential to know the special advantages and disadvantages of each concept to be able to choose the right strategy of treatment. Even getting the right diagnosis may be difficult since only about 2/3 of all cases go along with a positive microbiology. By the example of the infected hip prosthesis, the aim of the article is to give recommendations on the bases of current literature and our own experience. The strategy of treatment of infections used in our hospital is reported. PMID- 11398623 TI - [Experiences with the German Endoprosthesis Register]. AB - The German Endoprostheses Register started its documentation of primary and revision surgery on major joints in 1997. About 7000 primary arthroplasties and 1100 revisions on hip joints as well as 3000 primary arthroplasties and 250 revisions on knee joints are registered annually by 41 hospitals, which amounts to 60% in primary or 80% in revision surgery of the Swedish arthroplasty register. 95% of the revisions are performed on implants which have not been primarily operated on by the hospitals enrolled to the register, normally recruiting the classical "lost-to-follow-up-cases". Especially the removed implants show a conspicuous diversity of models. More than 10 specimens were removed in only 24 of 173 different types of hip-stems and 21 of 133 different types of hip-sockets. Thus, in order to cover not only a small number of types of implants by statistical analysis, large numbers of surgery have to be registered to yield case-numbers which can not be achieved by registers of small countries like the Scandinavian. Germany with approximately 1900 relevant clinical departments owns a slumbering potential that could easily be recruited. PMID- 11398624 TI - [Fractures of the femoral neck--results of an external quality assurance in 32,000 patients]. AB - Between Jan. 1st 1993 and Dec. 31st 1999 data was collected on 31.917 patients with fractures of the femoral neck in a statewide report card system in Westphalia-Lippe. Only 6.3% of patients were younger than 60 years. Ten percent of the population of Germany live in Westphalia-Lippe so that our data can be considered representative of Germany. A significant change was seen in the following indicators of quality: increase of operative treatment 1993 93.2%--1999 96.0% (p < 0.01 Chi Quadrat Test), decrease of mortality 1993 6.9%--1999 5.7% (p < 0.01 Chi Quadrat Test), decrease of cardiopulmonary complications 1993 11.2%- 1999 8.4% (p < 0.01 Chi Quadrat Test), decrease of mean length of stay 1993 33.5 days--1999 22.2 days (p < 0.01 one-way-ANOVA), decrease of pre-operative length of stay 1993 2.57 days--1999 1.86 days (p < 0.01 one-way-ANOVA), increase of operative treatment on weekends (p < 0.01 Chi Quadrat Test). 25% of the patients had post-operative complications. Intra-operative complications were rare (1.4%). Only 48.6% of internal fixations were performed on the day of admission. The most frequent operation is hemiarthroplasty and its frequency is still increasing. Total hip replacement was done 34.1% of all cases. More patients are transferred from primary hospitals to rehabilitation facilities following operative procedures: 1993 8.3%; 1999 27.2%. PMID- 11398625 TI - [Is autologous chondrocyte implantation a technology suitable for ambulatory health care?]. AB - Statutory ambulatory health care services provided by physicians in Germany are specified by a joint committee of physicians and sickness funds, the Federal Committee of Physicians and Sickness Funds. To ensure that only effective interventions are covered, new medical technologies must demonstrate evidence of clinical effectiveness, cost-effectiveness and medical necessity. Due to increasing demands for reimbursement of autologous chondrocyte implantation, the Committee recently assessed this procedure. Autologous chondrocyte implantation (ACI) is a surgical procedure to reestablish the chondral layer in the knee following trauma. Such injuries may subsequently lead to osteoarthritis, and may eventually require joint replacement. Although the U.S. Food and Drug Administration (FDA) approval of ACI limits its usage to treatment of specific knee defects, other indications, such as for chondromalacia patellae, are promoted by proponents of the technology. The Committee conducted a thorough literature review and examined the status of ACI in other health care systems. Reimbursement of this technology was declined for the following reasons: the effectiveness of ACI has not yet been established in long term, comparative studies, and the procedure involves a type of operation not suitable for the ambulatory health care sector. Other public health insurance systems, such as in Switzerland and Canada, have not yet introduced funding for ACI, and an assessment is currently being conducted by the UK National Institute for Clinical Excellence (NICE). Autologous chondrocyte implantation is a technology that will compete with various other new interventions to correct chondral defects of the knee. It is essential that the treatment effect of ACI be demonstrated in clinically sound, comparative trials before this technology is introduced into the public health care system. PMID- 11398626 TI - [The LORAS project and quality assurance. In four years from input- to outcome oriented financing in public health. 2: LORAS project outcome parts 1 & 98]. AB - This series of three articles is a summary of the operations, findings and results of the hospital reform projects in the Canton of Zurich, termed LORAS. With the aid of the LORAS project within four years Zurich hospitals have been transformed. Whereas they used to adhere to input-oriented covering of deficits they now operate with outcome-oriented prospective financing of output. Part 1 describes the whole project. Part 2 focuses on the development of outcome measurement. Part 3 finally describes the implementation of the outcome measurement in the canton of Zurich. PMID- 11398627 TI - [The first treatment for bone fractures and dislocations. From the series on "First Medical Treatment" occasioned by the Central Committee for Medical Education in Prussia (Winter Semester 1904-05)]. PMID- 11398628 TI - Losing my father, one day at a time. PMID- 11398629 TI - Foreign exchanges. PMID- 11398630 TI - Meeting of the minds. PMID- 11398631 TI - Ruptured intracranial aneurysms. A review of diagnosis and management. AB - Despite decades of aggressive efforts to improve the outcome from ruptured intracranial aneurysms, subarachnoid hemorrhage (SAH) still carries high morbidity and mortality rates. Aneurysmal subarachnoid hemorrhage continues to be a frightening and poorly understood condition, contributing to delays in diagnosis and compromising patient care. Prompt diagnosis followed by aggressive treatment represents the best available method to improve patient outcome. Optimal results depend on rapid medical stabilization of the patient, early aneurysm repair to prevent rebleeding, and prevention of the recognized complications of subarachnoid hemorrhage. This review summarizes recommendations for managing the patient with a ruptured intracranial aneurysm. PMID- 11398632 TI - Parkinson's disease. An overview of treatments and research. PMID- 11398633 TI - Minor consent to treatment. PMID- 11398634 TI - Creating the mouse that roared. PMID- 11398635 TI - Chromoblastomycosis: clinical and mycologic experience of 51 cases. AB - This is a study of 51 cases of chromoblastomycosis detected in a 17-year period, all of which were clinically and mycologically proven by direct examinations, cultures and biopsies. The therapeutic results of the various treatments used are reported. Most cases were males (36 of 51; 70%), the mean age was 35 years and farmers predominated (74%); the most frequent lesions were in the lower limbs (54%). Major clinical presentations were nodular (41%) and verrucous (26%). The principal aetiologic agent isolated was Fonsecaea pedrosoi (90%). Overall results of the various treatments were as follows: 31% were cured, 57% improved and 12% failed. The best results were obtained with cryosurgery for small lesions, with itraconazole for large ones, and in some cases the combination of both treatments. PMID- 11398636 TI - Transmission of Trichosporon asahii oesophagitis by a contaminated endoscope. AB - Two cases of oesophageal trichosporonosis due to a suspected nosocomial infection are reported. Both the patients were immunocompetent and had undergone an endoscopic examination on the same day. Six strains of Trichosporon were isolated: three strains from the oesophageal biopsy of the first patient, one strain from the endoscopic forceps, one from the air in the endoscopy room, and one from the oesophageal biopsy of the second patient. The nosocomial nature of the infection and the role of the endoscopic forceps in transporting the micro organism was suspected, but the morphology and physiology of the isolated strains did not confirm such hypothesis. To elucidate the nature of the infection and the genetic similarities of the strains isolated, all strains were typed with RFLPs of the rDNA fragment and with RAPD. The results of RAPD using primer (GTG)5 (GACA)4, M13 core sequence, and the 15-mer oligonucleotide GAGGGTGGXGGXTCT indicated the molecular identity of three strains supporting the hypothesis concerning a transport of the aetiological agent from the first patient to the second and that the carrier was the forceps of the endoscopic device. PMID- 11398637 TI - Chlamydospore formation on Staib agar. Observations made before Candida dubliniensis was described. AB - When routinely using Staib agar to detect Cryptococcus neoformans in AIDS patients by the brown colour effect of its colonies, rough-looking colonies of a questionable variety of Candida albicans were also found. Microscopically, these colonies consisted of pseudohyphae with abundant masses of chlamydospores. However, the colonies of C. albicans were smooth-edged and formed by round-oval blastospores only. Such observations were made during the mycological supervision of 36 cryptococcosis cases during the 1987-94 period. All these questionable cultures of Candida spp. were discarded. However, because the corresponding photographs of and records on such strains were found to be identical with those recently published by molecular biologists under the title 'Chlamydospore formation on Staib agar as a species-specific characteristic of Candida dubliniensis' [Staib, P. & Morschhauser, J. (1999) Mycoses 42, 521-524], the present communication presents a report on such observations in a representative and exemplary case of an AIDS patient. PMID- 11398638 TI - Adhesion of Candida parapsilosis to epithelial and acrylic surfaces correlates with cell surface hydrophobicity. AB - We investigated in vitro adherence of 24 isolates of Candida parapsilosis and 12 isolates of Candida albicans with regard to their relative cell-surface hydrophobicity (CSH), adherence to human buccal epithelial cells (BEC) and acrylic surfaces. There was no significant interspecies difference in the relative adherence of C. parapsilosis and C. albicans isolates to BEC, although the former demonstrated a tendency for increased adherence. However, a significant intra-species variation in adherence among isolates of C. parapsilosis (P < 0.0001) to BEC, but not of C. albicans was noted. The superficial isolates of C. parapsilosis demonstrated a higher avidity (33%) to BEC than the systemic isolates. On regression analysis a significant positive correlation between C. parapsilosis adherence to BEC and denture acrylic surfaces was noted (r = 0.45, P = 0.02). Similarly, buccal cell adherence correlated strongly with CSH of C. parapsilosis (r = 0.63, P = 0.0008). These results shed further light on the intimate relationship between adherence and CSH in candidal colonization and imply that both C. parapsilosis and C. albicans are equally potent in colonizing mucosal surfaces with respect to the attributes investigated. PMID- 11398639 TI - Impact of endpoint definition on the outcome of antifungal susceptibility tests with Candida species: 24- versus 48-h incubation and 50 versus 80% reduction in growth. AB - The growth inhibition patterns of 764 clinical yeast isolates, in response to amphotericin B, flucytosine, itraconazole and fluconazole, were studied in order to determine the frequency of trailing growth and any impact this, as well as 24- or 48-h incubation periods, may have on minimum inhibitory concentration (MIC) results. A broth microdilution method following National Committee for Clinical Laboratory Standard No. M27A recommendations was used. Trailing growth was observed mainly with azoles. Furthermore, over 98% of isolates exhibiting a trailing effect at 24 h with fluconazole and itraconazole were either Candida albicans or Candida tropicalis. When comparing 24- and 48-h IC50 values, discrepancies were observed with itraconazole and fluconazole, respectively, in 18 and 11% of C. albicans and 24 and 30% of C. tropicalis. When comparing IC50 and IC80 values at 24 h, discrepancies were again essentially seen with itraconazole and fluconazole, respectively, in 11 and 10% of C. albicans, and 17 and 27% of C. tropicalis. In susceptibility tests performed with a microdilution method and read spectrophotometrically, 48-h IC80 values result in an unlikely high number of resistant isolates, indicating that a 24-h incubation and a 50% reduction in optical density may correlate better with clinical outcome. PMID- 11398640 TI - The utility of serology in diagnosing candidosis in non-neutropenic critically ill patients. AB - This study was carried out to evaluate the utility of serological tests in the diagnosis of candidal infections in non-neutropenic critically ill patients. A prospective study was carried out in a 10-bed general intensive care unit; all patients with at least one organic sample with Candida spp. were included. A therapeutic-diagnostic algorithm was designed, and patients were treated or not, according to a classification. Blood samples were taken, and serological tests included: antigenaemia detection using two different commercial latex kits (Cand Tec and Pastorex) and antibody detection by indirect haemagglutination (IHA) and indirect germ tube immunofluorescence (IFA). A total of 56% of antibody tests (IHA 45% and IFA 64%) and 26% of antigen tests (Cand-Tec 36% and Pastorex 17%) were positive. The sensitivity and specificity of these tests with respect to systemic candidosis were 37% and 78%, respectively, for antibodies, and 0% and 90% for antigens. There was statistical significance for mortality and low levels of antibodies; Candida glabrata was detected by IFA and Candida tropicalis by Cand-Tec. Serological tests may help to define the prognosis of these patients and to support the detection of specific Candida species. PMID- 11398641 TI - The aetiological agents of tinea capitis in Zaragoza (Spain). AB - Between 1977 and 1997, 190 cases of tinea capitis were observed. The age groups most commonly infected were 1-10 years old. The following dermatophytes were isolated: Microsporum canis (119), Trichophyton mentagrophytes (57), Trichophyton tonsurans (seven), Trichophyton verrucosum (three), Trichophyton violaceum (one), Trichophyton schoenleinii (one), Trichophyton soudanense (one) and Microsporum audouinii (one). PMID- 11398642 TI - Case report. Rhinocerebral zygomycosis. AB - A case of rhinocerebral zygomycosis treated with liposomal amphotericin B is described. PMID- 11398643 TI - Cerebriform colonies of Paracoccidioides brasiliensis isolated from nine-banded armadillos (Dasypus novemcinctus) at room temperature. AB - Twelve isolates of Paracoccidioides brasiliensis generated cerebriform colonies at room temperature on potato glucose agar slants (PDA). These isolates contained abundant chlamydospores and yeast-like cells and are a subset of the 65 isolates obtained from nine-banded armadillos (Dasypus novemcinctus). They grew as a yeast form with typical multiple buddings at 37 degrees C on brain heart infusion agar supplemented with 1% glucose. After replating on PDA and culturing at room temperature for 2 months, the mutants appeared as cottonous colonies, which indicated that the morphological characteristics were unstable. PMID- 11398644 TI - Supplemental insurance: Medicare's accidental stepchild. AB - The majority of Medicare beneficiaries supplement the basic Medicare benefit package with additional insurance. This article reviews the literature on Medicare supplemental insurance. Supplemental insurance plays a significant role in protecting Medicare beneficiaries from financial risk. The two major sources of coverage for beneficiaries--former employers and individual purchase--differ in benefit structure and characteristics of policy holders. Employer-sponsored policies tend to provide broader coverage with more cost sharing than individually purchased policies, and holders of employer policies tend to be younger, wealthier, healthier, and better educated. Supplemental insurance policies have been shown to be associated with higher Medicare expenditures, but there is no consensus on the cause of the higher expenditures. Some studies attribute the increase to adverse selection of policies; other studies point to the moral hazard effect of insurance. PMID- 11398645 TI - CEO perceptions of competition--and strategic response in hospital markets. AB - Physician-organization integration (POI) has emerged as a key issue for hospitals and health systems seeking to improve the quality and cost-effectiveness of care. Although competition and managed care are often cited as primary market drivers of the adoption of POI strategies, prior research has shown only weak associations between these market attributes and POI. This article argues that the role of key organizational decision makers has not been adequately accounted for in explaining strategic change. The study examines the role of hospital CEO perceptions of competition in predicting the adoption of five different approaches to POI. CEO perceptions of general market competition are explained by a combination of market and organizational attributes. Furthermore, when controlling for objective characteristics of the environment and organization, CEO perceptions of competition have consistent, statistically significant associations with four of five measures of POI examined. PMID- 11398646 TI - Institutional and economic influences on the adoption and extensiveness of managerial innovation in hospitals: The case of reengineering. AB - In the recent past, a number of managerial innovations--including product line management, total quality management, and reengineering--have swept through the hospital industry. Given their pervasiveness and their cost, understanding the mix of factors that influences their adoption is of theoretical interest and practical relevance. The research reported here focuses on this general question by examining influences on the adoption and extensiveness of a particular managerial innovation, hospital reengineering. The results suggest that while economic and institutional factors have influenced the adoption and extensiveness of hospital reengineering, institutional forces play a more important role. The greater influence of institutional forces may be attributed to the high degree of uncertainty in health care, the causal ambiguity of the innovation, and the anticipatory actions of hospitals attempting to position themselves in a rapidly changing environment. PMID- 11398647 TI - Inequality in America: the contribution of health centers in reducing and eliminating disparities in access to care. AB - Reducing and eliminating health status disparities by providing access to appropriate health care is a goal of the nation's health care delivery system. This article reviews the literature that demonstrates a relationship between access to appropriate health care and reductions in health status disparities. Using comprehensive site-level data, patient surveys, and medical record reviews, the authors present an evaluation of the ability of health centers to provide such access. Access to a regular and usual source of care alone can mitigate health status disparities. The safety net health center network has reduced racial/ethnic, income, and insurance status disparities in access to primary care and important preventive screening procedures. In addition, the network has reduced low birth weight disparities for African American infants. Evidence suggests that health centers are successful in reducing and eliminating health access disparities by establishing themselves as their patients' usual and regular source of care. This relationship portends well for reducing and eliminating health status disparities. PMID- 11398648 TI - [Head transplantation?]. PMID- 11398649 TI - [New thrombolytic agents. Thrombin and factor Xa inhibitors]. PMID- 11398650 TI - [Thyroid diseases as a cause of skin changes and hair loss]. PMID- 11398651 TI - [Description and therapy of hearing disorders]. PMID- 11398652 TI - [Diphtheria]. PMID- 11398653 TI - [Colloidal silver in infections?]. PMID- 11398654 TI - NHS Direct. Dial M for ... medical advice. PMID- 11398655 TI - Working lives. The big shot. Interview by Barbara Millar. PMID- 11398656 TI - Organisational development. Reigning cats and dogs. AB - Research into the culture of two trusts involved in a merger revealed significant differences. This provided a valuable framework for developing the new organisation and convinced the board of the need for change. Annual appraisals for all staff, 360 degrees feedback for senior managers and clinicians and more comprehensive selection procedures have been introduced. PMID- 11398657 TI - Data briefing. Caesarean births. PMID- 11398658 TI - e-novation: managing with new technology. Live wires. PMID- 11398659 TI - e-novation: managing with new technology. Mind the gap. PMID- 11398660 TI - e-novation: managing with new technology. Mad scramble. PMID- 11398661 TI - e-novation: managing with new technology. At the crossroads. PMID- 11398662 TI - Breeding preferences of Anopheles culicifacies in the rice agro-ecosystem in Kheda district, Gujarat. AB - Breeding preferences of Anopheles culicifacies, a principle malaria vector, in the plains of India was studied in the rice agro-ecosystem of Kheda district in central Gujarat. Extensive breeding of this species was found in the rice field channels (20.83 per cent) and in rice fields (5.32 per cent). However, rice nurseries (0.91 per cent) and post-harvested rice fields (2.51 per cent) were less preferred. The species was found in abundance in newly transplanted rice fields and during early months of rice cultivation with a peak prevalence in the non-monsoon (Rabi) season. The breeding of An. culicifacies was inversely proportional and negatively correlated (r = -0.868; p < 0.05) with the height of the plants, whereas it showed a positive correlation (r = 0.779; p < 0.05) with the distance between plants. Rice fields near the villages supported maximum breeding of An. culicifacies (48 per cent) followed by the rice fields, 0.5 to one km away from the human habitation. Co-efficient of association (C8 index) revealed a positive association of the species with An. annularis, An. pallidus, An. subpictus and Cx. quinquefasciatus. However, it was negatively associated with An. nigerrimus, Cx. tritaeniorhynchus and Cx. vishnui sub groups. PMID- 11398663 TI - Effect of arteether on electrocardiogram in the patients of falciparum malaria--a preliminary study. AB - The effect of intramuscular arteether (150 mg daily for 3 days) on electrocardiogram was studied in 16 patients having falciparum malaria. This included three patients with cerebral malaria, three patients with jaundice (S. bilirubin > 3 mg per cent) and ten patients with uncomplicated malaria. Five patients had tachycardia prior to arteether treatment. The mean RR interval before starting the treatment was 0.59 sec which increased progressively--0.60, 0.68 and 0.69 sec on Day 1, 2 and 3 respectively after starting the treatment. There was no significant difference statistically in the mean corrected QT interval (p > 0.05), PR interval (p > 0.05), QRS duration (p > 0.05) and diastolic BP (p > 0.05) before starting and at the end of treatment on Day 3. One patient had developed first degree heart block (PR 0.24), while another patient had prolonged QTc (0.48 sec) on Day 3. Subsequent ECG of both these patients was normal on Day 5. The profile of ECG changes was same in the patients of uncomplicated and complicated malaria. Absence of any significant effect on BP and ECG changes precludes the significant effect of arteether on the cardiovascular system when compared to quinine which may cause hypotension, arrhythmia and QTc prolongation. PMID- 11398664 TI - Field studies on the sensitivity and specificity of an immunochromatographic test for the detection of Plasmodium falciparum malaria in tribal areas of Orissa. AB - A rapid immunodiagnostic test developed by an Australian Biotechnology company for the diagnosis of Plasmodium falciparum in the peripheral blood has been evaluated in the field for its sensitivity, specificity and efficacy in comparison to microscopic examination. The results showed that the tests sensitivity, specificity and efficacy were 98.2, 96.9 and 97.5 per cent respectively. The positive and negative predictive values of the test were 96.4 and 98.4 per cent respectively. The test when compared to the conventional microscopy did not show any statistically significant difference suggesting that the two diagnostic methods are equally good. The test performed did not show cross-reactions with other parasite species. It is a simple and rapid field diagnostic method, which does not require any expensive laboratory equipment or skilled personnel. PMID- 11398665 TI - Raised serum thiobarbituric acid reactive substance levels in malaria. AB - To study the extent of serum lipid peroxidation in malaria, 62 patients of falciparum malaria (18 uncomplicated and 44 complicated), 15 patients of vivax malaria and 25 healthy controls were enrolled in this study. The extent of serum lipid peroxidation was evaluated by estimating serum thiobarbituric acid reactive substances (TBARS) colorimetrically. The mean serum TBARS levels were 1.5 +/- 0.29, 1.21 +/- 0.2 and 3.58 +/- 1.35 nmol/ml in controls, vivax malaria and falciparum malaria patients respectively. The TBARS level was significantly more in complicated falciparum malaria patients (4.2 +/- 1.03 nmol/ml) than uncomplicated falciparum malaria patients (2.01 +/- 0.61 nmol/ml). The TBARS level was also more in patients who died (4.82 +/- 0.64 nmol/ml) when compared to the survivors (2.92 +/- 1.05 nmol/ml). PMID- 11398666 TI - Malaria investigation in District Jodhpur, Rajasthan, during the summer season. AB - Studies were carried out in District Jodhpur of the Thar region of Rajasthan. Epidemiological investigation revealed high slide positivity rate in the canal irrigated area (54.5 per cent), sand dunes area (67.54 per cent), stone quarry area (26.66 per cent) and in the desert plain area (41.5 per cent). Similarly, slide falciparum rates were 7.10, 4.38, 6.66 and 5.6 per cent respectively. Entomological studies showed An. stephensi and An. culicifacies as major species and their densities ranged between 2 to 14.58 and 0 to 0.9 pmh respectively. Resistance in malaria vectors to insecticides, poor surveillance and suppressive treatment of cases appear to be the factors for persistent transmission in the study area. PMID- 11398667 TI - Susceptibility status of Culex quinquefasciatus larvae to fenthion in Delhi--a note on the possible development of resistance. PMID- 11398668 TI - A study of human genetic markers in Mewat region, Gurgaon, Haryana. PMID- 11398669 TI - Susceptibility status of An. fluviatilis and An. culicifacies to DDT, deltamethrin and lambdacyhalothrin in District Nainital, Uttar Pradesh. PMID- 11398670 TI - Serial ParaSight-F test in patients with severe malaria. PMID- 11398671 TI - [Radicality criteria in head and neck tumors. Histopathologic presuppositions]. PMID- 11398672 TI - [Molecular markers study in pTNM of squamous carcinoma of the head and neck]. PMID- 11398673 TI - [Microsatellite molecular analysis of the resection margins and laterocervical lymph nodes in the planning of complementary postoperative radiotherapy in early squamous carcinoma (stage I and II) of the head and neck]. PMID- 11398674 TI - [The problem of surgical resection margin of the oral cavity-oropharynx, cervical hypopharyngeal esophagus and facial maxillary]. PMID- 11398675 TI - [Correlation between resection margin status and local recurrence in surgery of laryngeal carcinoma]. PMID- 11398676 TI - [Significance of surgical radicality as presupposition of therapeutic success]. PMID- 11398677 TI - [Prospects and therapeutic decisions in the light of biological findings in laryngeal cancer]. PMID- 11398678 TI - [Multicenter survey through a questionnaire on the prognostic value of surgical resection margin in head and neck tumors]. PMID- 11398679 TI - [Study of click-induced vestibule-sternocleidomastoid reflex recorded with surface electromyography]. AB - The otolithic organs serve to control posture and maintain the visual objective during linear accelerations. Recent experimental research in man and animals has suggested that it may be possible to explore the function of these organs by studying the vestibule-sternocleidomastoid reflex induced by high intensity clicks. The aim of the present work was to identify the parameters which are most reproducible in normal subjects and, therefore, best suited to this purpose. The instrument normally used to study evoked auditory potentials (ERA) was used to perform an electromyographic recording at the surface of the sternocleidomastoid muscle (SCM) and then average the electromyographic response to short clicks (0.1 ms) introduced through the headphones. In normal subjects clicks at an intensity of 85-100 dB HL evoked a reproducible electromyographic response in the venter of the SCM muscle with an average latency of 8.75 ms. The latency parameter proved to present the most suitable characteristics for possible clinical use. The possibility of using this method to study human vestibular lesions is certainly enticing although further research is needed to better characterize the precise site of origin for the vestibule-sternocleidomastoid reflex. PMID- 11398680 TI - [Epistaxis: emergency treatment approach]. AB - After a brief summary of the causes of severe epistaxis and the surgical choices available for treatment, the authors present their own cases study. Starting from a brief description of the anatomy involved in the system of arterial irrigation of the nasal cavities, they focus on two fundamental concepts: the basis for a proper topographic picture of the site of bleeding and the success of surgery. First and foremost is the emergence of two arterial branches from the sphenopalatine formen; this differs from what is asserted in the classical treatise on anatomy and in line with what has been stated in the Nomina Anatomica, approved during the X World Congress on Anatomy (Tokyo 1975). Secondly, focus is placed on the existence of two important anastomotic systems in the arterial circulation of the nasal cavities: the one between nasal septum and anterior-posterior ethmoid arteries and the one between the lateral-posterior nasal and nasoseptal arteries. In the literature the failure of surgery in treating severe epistaxis is explained by the onset of supplementary anastomotic circulation. The indications for surgical treatment of epixstaxes are: copious bleeding in hospital or after the packing has been removed; underlying pathology or general status which has been aggravated by the packing; patient which has already been staunched several times. The results of 31 epistaxis procedures are presented: 8 (25.81%) upper and 23 (74.19%) posterior. The upper epistaxes were treated as follows: 3 cases with anterior ethmoid ligature, two of which failed; 2 cases with combined ligature of the anterior and posterior ethmoid; 3 cases of successful septoplasty. The posterior epistaxes were treated as follows: 11 cases of ligature of the lateral-posterior nasal artery, 2 of which failed; 12 cases of successful combined ligature of lateral-posterior nasal and nasoseptal arteries. These results confirm that it is possible to effectively resolve surgically severe epistaxis using microsurgery or transnasal endoscopy based on in depth knowledge of the complex architecture of the arterial circulation within the district of the paranasal sinuses, possible anatomic variations and collateral circulation. PMID- 11398681 TI - [Prophylactic postoperative radiotherapy in the treatment of supraglottic tumors]. AB - The appropriateness of treatment of supraglottic carcinoma with post-operative radiotherapy is still a controversial issue. The purpose of the present work has been to define the effectiveness of post-operative radiotherapy in the treatment of supraglottic carcinomas and to discuss, on the basis of the data reported in the literature, the usefulness of combined surgery-radiotherapy. The study involved 97 subjects suffering from spinocellular carcinoma of the laryngeal vestibule (95 males, 2 females; average age: 58; age range: 38-89 years) who underwent horizontal supraglottic laryngectomy together with bilateral laterocervical lymph node dissection. Of these patients 35 (36%, group A) had undergone a cycle of prophylactic radiotherapy (60-70 Gy in fractions of 2 Gy/die, administered with two side fields), after surgery. The remaining 62 cases (64%, group B) did not have any additional therapy after surgery. For both groups the overall actuarial survival and 5-year corrected survival rates were calculated; statistical significance was calculated using the Wicoxon test. For group A the overall actuarial survival and corrected actuarial survival rates were, respectively, 74% and 90%. For group B these rates were, respectively 61% and 80%. A statistical comparison of both parameters did not show any statistically significant difference (p = 0.2 and 0.4, respectively). In reference to tumor extension, established both on the basis of clinical (T) and surgical-anatompatholgical (pT) findings, no significant differences were encountered between the overall actuarial survival and corrected actuarial survival rates for the two groups of patients (p > 0.05). In NO patients significantly higher overall actuarial survival rate in those patients who underwent the combined surgery-radiotherapy treatment than in those treated with surgery alone (p = 0.01) was seen. This was not, however, confirmed by the corrected actuarial survival rates and comparative analysis of the groups for surgical staging of the lymph node metastases (p > 0.05). Most likely the different behavior seen in the two groups depends on possible errors in the clinical staging of N due to the presence of reactive lymphadenitis and micrometastases. In conclusion, the data derived from the present research indicate that post-operative radiotherapy, performed as prophylaxis in cases undergoing supraglottic laryngectomy does not yield any statistically significant improvement in prognosis. Therefore, the radiotherapy association should be ruled out in those patients where the oncological radicality of the surgery is reasonably certain. It can possibly be considered as integration to surgery, only in those cases where radicality is in doubt. PMID- 11398682 TI - [Cricohyoidoepiglottopexy : deglutition in 44 cases]. AB - The present work reviews the authors personal experience and applies a meta analysis of the data in the literature (approximately 800 cases) to better focus on the opportunities and difficulties involved in the Majer-Piquet technique. Particular attention is focused on the problems involved in deglutition. The study involves 44 patients who underwent cricohyoidoepiglottopexy (CHEP) between 1989 and 1998. Given that surgical and rehabilitation techniques have been refined over the years, to better analyze the functional results, the data were separated into two subsequent periods. Of the 23 patients who underwent surgery between 1989 and 1992 (group I), 6 were benefited from bilateral preservation of the arytenoids while this was only possible in two of the 21 cases operated in the second period from 1993 to 1998 (group II). Functional rehabilitation was started earlier in the second group and was routinely monitored with digital viedeofluorography. The average recovery time was 34 days for group I and 27 days for group II. The tracheostomy closed in an average 91 days vs. the 13 days found by the meta-analysis of the literature. The authors normally leave the tracheostoma in place for a long time, even when closed with an easily removed plug, and only perform plastic surgery when the patient has shown stable deglutition for several weeks. The naso-gastric tube was removed from both groups of patients after an average 16 days while the review of the literature shows an average 21 days. On the whole the authors record good deglutition in 41 of the 44 cases (93.18%) with adequate deglutition in the remaining 3 cases. Likewise the literature reports good deglutition in 86.4% of the cases. The only difference found between the two groups of patients was a quicker recovery in the second group; there were no qualitative differences. The results are described and discussed. In conclusion, the present experience places CHEP in favorable light as long as the limits inherent to the method are recognized. However, these limitations relegate the technique to a "sporadic" role and the review of the literature appears to confirm this attitude. PMID- 11398683 TI - [Informed consent. Proposal of a method for ORL]. PMID- 11398684 TI - [The 26th National Meeting of the A.I.O.L.P. -- Salsomaggiore Terme (Pr), April 28-30, 2000]. PMID- 11398685 TI - Intermittent administration of furosemide or continuous infusion in critically ill infants and children: does it make a difference? PMID- 11398686 TI - Approaches to suspected ventilator-associated pneumonia: relying on our own bias. PMID- 11398687 TI - Pharmacokinetic studies in patients on continuous renal replacement therapies. PMID- 11398688 TI - Computerised tomography scan imaging in acute respiratory distress syndrome. AB - Computerised tomography (CT) is being used with increasing frequency in acute respiratory distress syndrome (ARDS) patients. This brief review will discuss some of the clinical insights that a CT scan can offer. A large number of CT scan studies have provided new insights into the pathophysiology of ARDS and of mechanical ventilation, and are particularly focused on the recruitment derecruitment phenomenon. To this end, newer fast CT scan technology promises a dynamic, rather than a static view of lung ventilation. PMID- 11398689 TI - Usefulness of a strategy based on bronchoscopy with direct examination of bronchoalveolar lavage fluid in the initial antibiotic therapy of suspected ventilator-associated pneumonia. AB - OBJECTIVES: To evaluate (a) the routine accuracy of bronchoalveolar lavage by direct examination (BAL-D) in diagnosing ventilator-associated pneumonia (VAP), and (b) the impact of a diagnostic strategy including clinical judgment, bronchoscopy, and BAL-D on the initial diagnosis and appropriateness of treatment when VAP is suspected. DESIGN AND SETTING: Prospective cohort study in two academic ICUs in Paris, France. PATIENTS AND PARTICIPANTS: Mechanically ventilated patients with suspected VAP underwent bronchoscopy with BAL and protected specimen brush (PSB). BAL-D results were available within 2 h, BAL on culture and PSB results after 24 h, and antibiotic susceptibility after 48 h. At each step in the strategy the senior and the resident in charge of the patient were asked their diagnosis and their therapeutic plan on the basis of presently available data. Definite diagnosis of suspected VAP was based on histology, appearance of cavitation, positive pleural fluid culture, results of PSB and BAL culture, and follow-up. MEASUREMENT AND RESULTS: A total of 110 episodes of suspected VAP were studied; 94 definite diagnoses were made (47 VAP, 47 no VAP). Using a threshold 1% of infected cells, BAL-D discriminated well between patients with and those without VAP (sensitivity 93.6%, specificity 91.5%, area under the receiver-operating characteristic curve 0.953). The senior clinical judgment was correct in 71% cases. It was correct in 78% and 94% of cases after airway visualization and BAL-D findings, respectively. After BAL-D the positive and negative predictive values in diagnosing VAP were 90% and 98%, respectively. However, the therapeutic plan was correct in only 65% using clinical judgment (15 untreated patients, 3 ineffective treatment, 15 useless treatment), 66% using airway visualization (14 untreated VAP, 4 ineffective treatment, 14 useless treatment), and 88% using BAL-D results (1 untreated patients, 6 ineffective, 4 useless), according to definite diagnosis and final antibiotic susceptibility testings. CONCLUSIONS: A strategy based on bronchoscopy and BAL-D generally leads to a rapid and appropriate treatment of nosocomial pneumonia in ventilated patients. PMID- 11398690 TI - Closed system endotracheal suctioning maintains lung volume during volume controlled mechanical ventilation. AB - OBJECTIVE: A closed suction system (CS) maintains connection with the mechanical ventilator during tracheal suctioning and is claimed to limit loss in lung volume and oxygenation. We compared changes in lung volume, oxygenation, airway pressure and hemodynamics during endotracheal suctioning performed with CS and with an open suction system (OS). DESIGN: Prospective, randomized study. SETTING: Intensive care unit in a university hospital. PATIENTS: We enrolled ten patients, volume-controlled (VC) ventilated with a Siemens Servo 900 ventilator (PaO2/FIO2 192 +/- 70, PEEP 10.7 +/- 3.9 cmH2O). INTERVENTIONS: We performed four consecutive tracheal suction maneuvers, two with CS and two with OS, at 20-min intervals. During the suction maneuvers continuous suction was applied for 20 s. MEASUREMENTS AND MAIN RESULTS: We measured end-expiratory lung volume changes (delta VL), tidal volume (VTrt), respiratory rate (RR) and minute volume (VErt) by respiratory inductive plethysmography; arterial oxygen saturation (SpO2), airway pressure and arterial pressure (PA). Loss in lung volume during OS (delta VL 1.2 +/- 0.7 l) was significantly higher than during CS (delta VL 0.14 +/- 0.1 l). During OS we observed a marked drop in SpO2, while during CS the change was only minor. During CS ventilation was not interrupted and we observed an immediate increase in RR (due to the activation of the ventilator's trigger), while VTrt decreased, VErt was maintained. CONCLUSIONS: Avoiding suction-related lung volume loss can be helpful in patients with an increased tendency to alveolar collapse; CS allows suctioning while avoiding dramatic drops in lung volumes and seems to be safe during the VC ventilation setting that we used. PMID- 11398691 TI - Equal aspiration rates in gastrically and transpylorically fed critically ill patients. AB - OBJECTIVES: To determine the difference in aspiration rates between gastrically and transpylorically fed patients in the intensive care unit. DESIGN: A prospective controlled study of critically ill patients randomized to receive either a gastrically placed feeding tube or a transpylorically placed feeding tube. SETTING: University teaching hospital's medical intensive care unit. The study was conducted over 14 months. PATIENTS: Fifty-four critically ill subjects (with an overall 40% mortality) with similar baseline age, severity of illness, and nutritional needs requiring enteral nutrition, with 51 completing the study. INTERVENTIONS: All feeds were tagged with technetium-99m radiolabeled sulfur colloid, and the pulmonary secretions or lungs of each patient were scanned on a daily basis to determine whether aspiration had occurred. Patients were fed according to their assigned tube placement which was verified daily by continuous electromyography. MEASUREMENTS AND RESULTS: Of 27 gastrically fed patients 2 (7%) had evidence of scanned feed in pulmonary secretions or the lung, compared to 3 of 24 (13%) transpylorically fed patients (n.s.). Clinical suspicion of aspiration was insensitive and detected only 60% of isotopically documented aspirations with a positive predictive value of 27%. CONCLUSION: There was no difference in aspiration rates between gastrically and transpylorically fed critically ill patients. PMID- 11398692 TI - Pharmacokinetics of ciprofloxacin in ICU patients on continuous veno-venous haemodiafiltration. AB - OBJECTIVES: To investigate the pharmacokinetics of intravenous ciprofloxacin 200 mg every 8 h in critically ill patients on continuous veno-venous haemodiafiltration (CVVHDF), one form of continuous renal replacement therapy (CRRT). DESIGN AND SETTING: Open, prospective clinical study in a multidisciplinary, intensive care unit in a university-affiliated tertiary referral hospital. PATIENTS: Six critically ill patients with acute renal failure on CVVHDF. INTERVENTIONS: Timed blood and ultrafiltrate samples were collected to allow pharmacokinetics and clearances to be calculated of initial and subsequent doses of 200 mg intravenous ciprofloxacin. CVVHD was performed with 1 l/h of dialysate and 2 l/h of predilution filtration solution, producing 3 l/h of dialysis effluent. The blood was pumped at 200 ml/min using a Gambro BMM-10 blood pump through a Hospal AN69HF haemofilter. MEASUREMENTS AND RESULTS: Ten pharmacokinetic profiles were measured. The CVVHDF displayed a urea clearance of 42 +/- 3 ml/min, and removed ciprofloxacin with a clearance of 37 +/- 7 ml/min. This rate was 2-2.5 greater than previously published for ciprofloxacin in other forms of CRRT. On average the CVVHDF was responsible for clearing a fifth of all ciprofloxacin eliminated (21 +/- 10%). The total body clearance of ciprofloxacin was 12.2 +/- 4.3 l/h. The trough concentration following the initial dose was 0.7 +/- 0.3 mg/l. The area under the plasma concentration time curves over a 24-h period ranged from 21 to 55 mg.h l-1. CONCLUSIONS: Intravenous ciprofloxacin 600 mg/day in critically ill patients using this form of CRRT produced adequate plasma levels for many resistant microbes found in intensive care units. PMID- 11398693 TI - Acid-base alterations in heatstroke. AB - OBJECTIVE: To analyze the acid-base balance during heatstroke. DESIGN: Retrospective study. SETTING: Heatstroke Center, Makkah, Saudi Arabia. PATIENTS: Hundred nine consecutive heatstroke patients (mean age 55 +/- 12 years) with rectal temperature from 40 to 43.4 degrees C following exposure to hot weather. INTERVENTION: Arterial blood gases collected prospectively and analyzed using 95% confidence limits established by controlled experimental studies. Severity of heatstroke on admission assessed by Simplified Acute Physiology Score and Organ System Failure score. RESULTS: Metabolic acidosis was the predominant acid-base change followed by respiratory alkalosis (81 and 55% of the patients, respectively). The prevalence of metabolic acidosis (but not respiratory alkalosis) was significantly associated with the degree of hyperthermia: 63, 95 and 100% at 41, 42 and 43 degrees C, respectively (p < 0.0001). Patients with metabolic acidosis had a large anion gap (24 +/- 5). Arterial partial pressure of oxygen (PaO2), systolic blood pressure and Organ System Failure score were similar with or without metabolic acidosis. Although the acute physiology score was higher in patients with, than without, metabolic acidosis (15.7 +/- 3.7 vs 9.8 +/- 4.4, p < 0.001), there was no significant difference in neurologic morbidity and mortality (7.9 vs 1.1%, 5.6 vs 0%, p = 0.776 and 0.581, respectively). CONCLUSION: We conclude that metabolic acidosis is the predominant response in heatstroke. PMID- 11398694 TI - Critical illness polyneuropathy: clinical findings and cell culture assay of neurotoxicity assessed by a prospective study. AB - OBJECTIVE: First, to evaluate the role of typical intensive care-related conditions like sepsis, prolonged ventilation, drug effects and metabolic disorders in the pathogenesis of critical illness polyneuropathy (CIP); second, to investigate the possible significance of patient serum neurotoxicity assessed by an in vitro cytotoxicity assay with respect to CIP development. DESIGN: Prospective study. SETTING: Neurological intensive care unit. PATIENTS AND PARTICIPANTS: Twenty-eight patients who were on mechanical respiratory support for at least 4 days during a 21-month study period. RESULTS: Diagnosis of CIP was established by clinical and electrophysiological examination in 16 (57%) of 28 patients. Patients were investigated on days 4, 8 and 14 of mechanical ventilation. Two of 16 CIP patients had clinical signs of polyneuropathy at initial examination. Factors that correlated significantly with the development of CIP were: the multiple organ failure score on day 8 of ventilation, the total duration of respiratory support, the presence of weaning problems and the manifestation of complicating sepsis and/or lung failure. The in vitro toxicity assay showed serum neurotoxicity in 12 of 16 CIP patients. Electrophysiological investigations yielded false positive results of the toxicity assay in six patients (not developing CIP) and false negative results in four patients (developing clinical and electrophysiological signs of CIP). Statistical analysis did not reveal a significant correlation between the diagnosis of CIP and the finding serum neurotoxicity. CONCLUSION: The results support the hypothesis of a multi-factorial aetiopathogenesis of CIP. We observed serum neurotoxicity in the majority of CIP patients, indicating the possible involvement of a so far unknown, low-molecular-weight neurotoxic agent in CIP pathogenesis. PMID- 11398695 TI - Clinical features of patients with acute organophosphate poisoning requiring intensive care. AB - OBJECTIVES: To describe the clinical features of organophosphate poisoning (OPP), to evaluate the Acute Physiology and Chronic Health Evaluation (APACHE) II score as an alternative index for measuring OPP severity, and to assess cholinesterase levels for predicting successful weaning from mechanical ventilation (MV). DESIGN AND SETTING: Retrospective medical record review in a medical intensive care unit of an acute general hospital. PATIENTS: Twenty-three adults with OPP between 1995 and 1999. All cases were due to malathion poisoning. Muscarinic features were the predominant clinical manifestations (83%), followed by central nervous system (78%) and nicotinic manifestations (17%). RESULTS: MV was required by 74% of patients because of bronchial secretions (83%), altered conscious level (78%), pneumonia (78%), and flaccid paralysis (57%). Five patients (22%) had features of intermediate syndrome. ICU mortality was 13% and the mean ICU stay was 9.1 +/- 6.0 days. The mean APACHE II score was 17.4 +/- 7.4 and it correlated with mortality, severity of OPP, length of MV, and cholinesterase level. An APACHE II score of 26 or higher was predictive of mortality, with 95% sensitivity and 100% specificity. Threshold levels of serum and red blood cell cholinesterase for successful weaning from MV were 2,900 U/l and 7,500 U/l, respectively. CONCLUSIONS: The APACHE II score may be used as an alternative index of severity in patients with OPP; a score of 26 or higher is a good predictor of mortality. Cholinesterase levels are useful in predicting successful weaning of patients from MV. PMID- 11398696 TI - A prospective cohort study of 6-month mortality in a community hospital experiencing a gradual reduction in critical care services. AB - OBJECTIVE: To analyse the effect of reduction of critical care services on admissions, resource consumption and outcome. DESIGN: Observation outcome study with analysis of patient data collected prospectively during 1993, 1995 and 1997. SETTING: High dependency and intensive care unit (HDU/ICU) of a community hospital serving a population of 168,000. The number of beds decreased from 12 (1993), to 10 (1995) and to 8 (1997) with concomitant decrease in staff. PATIENTS AND PARTICIPANTS: Three patient cohorts admitted to the HDU/ICU during 1993, 1995 and 1997. MEASUREMENTS AND RESULTS: Admissions were classified into recovery room care or critical care admissions and stratified according to workload (Levels I IV). Illness severity scores of critical care admissions were recorded according to the APACHE II system. Mortality data were acquired from a national database. The total number of admissions to the unit did not change over the years. Length of stay decreased significantly over the years. Standardised mortality rates based on mortality within 30 days of discharge from the HDU/ICU were 1.17 (95% confidence interval 0.96-1.43) for critical care admissions during 1993, 0.86 (0.70-1.06) for 1995 and 0.98 (0.79-1.22) for 1997. Survival 180 days after discharge from the HDU/ICU did not differ significantly over the years. CONCLUSIONS: The results suggest that an excess of resources were used in critical care services during 1993 and 1995. Reduction of HDU/ICU beds by 30% from 7.1 to 4.8 beds/100,000 was not associated with increased 6-month mortality of the patients admitted. PMID- 11398697 TI - A comparison of post mortem findings with post hoc estimated clinical diagnoses of patients who die in a United Kingdom intensive care unit. AB - OBJECTIVE: To determine the accuracy of clinical diagnosis compared to post mortem findings in intensive care patients. DESIGN: A retrospective and blinded review of medical records. SETTING: A nine-bed combined high dependency and intensive care unit in a district general hospital in Gloucester, England. PATIENTS AND PARTICIPANTS: Ninety-seven patients who died on the intensive care unit and subsequently underwent post mortem examination. RESULTS: Suspected cause of death and main clinical diagnoses were determined and compared with findings at post mortem examination. All patients in whom a discrepancy was found had their cases reviewed to determine the significance of the discrepancy using the Goldman classification of post mortem discrepancies. Between June 1996 and May 1999 there were 1718 admissions with 252 deaths (14.7%), where 102 post mortem examinations were performed (40.5%). Five patients were not studied. Of the 97 patients, 4 (4.1%) had Goldman I discrepancies and 19 (19.6%) had Goldman II discrepancies. Complete agreement between pre and post mortem diagnosis was found in 74 (76.3%). Discrepancies fell into four main groups; unrecognised haemorrhage (7 patients), myocardial infarction (5), thromboembolic disease (5) and infectious complications (4). CONCLUSIONS: This study reveals that in an intensive care unit important diagnostic discrepancies were found in 19.6% of patients who underwent a post mortem examination. In a fifth of these (4.1%), survival may have been adversely affected. Haemorrhage was the most commonly missed diagnosis. Despite technological advances in intensive care medicine the post mortem examination continues to have an important role in auditing clinical practice and diagnostic performance. PMID- 11398698 TI - Continuous intravenous furosemide in haemodynamically unstable children after cardiac surgery. AB - OBJECTIVE: The commonly used continuous intravenous (i.v.) furosemide dosing schedule after cardiac surgery in children is largely empirical and may not be optimal. This may even be more marked in children after cardiac surgery who are haemodynamically unstable, and in whom transient renal insufficiency may occur. A study was performed to obtain an impression regarding which clinically applicable measures may be used to design a rational scheme for continuous i.v. furosemide therapy in children after cardiac surgery. SUBJECTS AND METHODS: Twelve paediatric patients (5F/7 M, age 0-33 weeks) post-cardiac surgery, who were to receive 3 days of continuous i.v. furosemide treatment, were included in an open study. Blood and urine samples were taken for furosemide, creatinine, and electrolyte levels, and fractionated urinary output was measured. Furosemide in blood and urine was measured using high performance liquid chromatography (HPLC). RESULTS: The mean starting dose of continuous i.v. furosemide was 0.093 (+/- 0.016) mg/kg per hour. The mean dose was increased to 0.175 (+/- 0.045) mg/kg per hour per hour on day 2, and changed to 0.150 (+/- 0.052) mg/kg per hour on day 3. Infusion rates were increased from day 1 to day 2 in ten cases, and decreased from day 2 to day 3 in three cases. Serum furosemide levels never exceeded ototoxic levels. The urinary furosemide excretion rate was inversely related to serum creatinine levels. CONCLUSIONS: This study extends the observation of the beneficial effects of continuous i.v. furosemide also to those children who are haemodynamically unstable after cardiac surgery. However, as the effects of furosemide are dependent on renal function, it can be hypothesised that the dosing schedule may be optimised. Contrary to the currently used dosage schedule in which the dose of furosemide is gradually increased over time, it may be more rational to start with a higher dose and adapt this dose (downward) guided by the observed effect (urine output). Because the infusion rate was increased to 0.2 mg/kg per hour in nine out of 12 patients on day 2 and was never increased further, this suggests that a starting rate of 0.2 mg/kg per hour may be optimal. PMID- 11398699 TI - How safe is non-bronchoscopic bronchoalveolar lavage in critically ill mechanically ventilated children? AB - OBJECTIVE: To assess the safety of non-bronchoscopic bronchoalveolar lavage (NB BAL) in critically ill mechanically ventilated children. SETTING: Paediatric intensive care unit in a tertiary children's hospital. METHODS: The data from 60 consecutive critically ill mechanically ventilated children who underwent NB-BAL was reviewed from November 1997 to December 1999. PRISM score prior to NB-BAL, observations at the time of NB-BAL and arterial blood gases, oxygenation index (OI), ventilator settings, haemodynamic variables and temperature taken at 1 h before, and 1 and 6 h after NB-BAL, were retrieved from the archived computerised database. RESULTS: Median age was 7 months (IQR 2.8-43 months) and median weight was 5.5 kg (IQR 4-14 kg). Four (7%) patients exhibited significant immediate complications, requiring escalation of respiratory or haemodynamic support. Forty two (70%) patients had complete data for calculation of OI; there was no significant change in median OI at 1 and 6 h after NB-BAL. However 5 (12%) of these patients experienced an increase in OI of between 10 and 45 at 1 h post NB BAL, which returned to baseline at 6 h post NB-BAL. Complications did not correlate with any of the available variables: baseline OI, PRISM score or with deterioration at the time of the procedure, although it was observed that four out of the six patients with baseline OIs of greater than 20 experienced complications. CONCLUSION: Non-bronchoscopic bronchoalveolar lavage in critically ill mechanically ventilated neonates and children is generally a well-tolerated procedure, but for some patients, in whom it was not possible to elucidate predictive factors, complications developed. All patients, particularly those with OIs of greater than 20, require careful monitoring during and after the procedure. PMID- 11398700 TI - Current management and outcome of tracheobronchial malacia and stenosis presenting to the paediatric intensive care unit. AB - OBJECTIVE: To identify factors associated with mortality and prolonged ventilatory requirements in patients admitted to our paediatric intensive care unit (PICU) with tracheobronchial malacia and stenosis diagnosed by dynamic contrast bronchograms. DESIGN: Retrospective review. SETTING: Tertiary paediatric intensive care unit. PATIENTS: Forty-eight cases admitted to our PICU over a 5 year period in whom a diagnosis of tracheobronchial malacia or stenosis was made by dynamic contrast bronchography (1994-1999). INTERVENTIONS: Conservative management, tracheostomy and long-term ventilation, surgical correction, internal or external airway stenting. MEASUREMENTS AND RESULTS: Recording of clinical details, length of invasive ventilation and appearance at contrast bronchography. Five groups of patients were defined: isolated primary airway pathology (n = 7), ex-premature infants (n = 11), vascular rings (n = 9), complex cardiac and/or syndromic pathology (n = 17) and tracheo-oesophageal fistulae (n = 4). The overall mortality was 29%. Median length of invasive ventilation in survivors was 38 days and in patients who died 45. Mortality was highest in the patients with complex cardiac and/or syndromic pathology (p = 0.039 Cox regression analysis) but was not related to any other factor. Patients with stenosis required a significantly longer period of ventilatory support (median length of ventilation 59 days) than patients with malacia (39 days). CONCLUSIONS: Length of ventilation and bronchographic diagnosis did not predict survival. The only factor found to contribute significantly to mortality was the presence of complex cardiac and/or syndromic pathology. However, patients with stenosis required longer ventilatory support than patients with malacia. PMID- 11398701 TI - Centralization of paediatric intensive care: are critically ill children appropriately referred to a regional centre? AB - OBJECTIVES: To evaluate the appropriateness of emergency referrals for inter hospital transfers by local physicians in hospitals without intensive care facilities to a regional tertiary paediatric intensive care unit (PICU). DESIGN: A prospective, descriptive study in a tertiary PICU and hospitals without paediatric intensive care facilities in and around the London area, UK. PATIENTS: All patients (< 18 years) referred for emergency admission to the PICU from district hospitals (n = 436) as well as those admitted through other modes of admission (n = 286) between 1 October 1998 and 30 June 1999 were prospectively studied. Admissions and transfers were deemed appropriate if the risk of mortality using the Paediatric Risk of Mortality (PRISM II) score was greater than 1%, and/or if the patient required a unique ICU-dependent therapy. Effectiveness was estimated using PRISM II derived observed-to-expected mortality ratio. Of the 436 emergency referrals 398 (91.3%) were retrieved and transported to the PICU. Of these, 38 referrals were thought to be inappropriate after telephone consultation and were not transferred. Of the emergency referrals 376 (94.4%) had a mortality risk greater than 1% or required an ICU-dependent therapy on admission day. Thus 86.2% (376/436) of the referrals and 94.4% (376/398) of transfers were considered appropriate. The PRISM II derived standardized mortality rate was 0.694 (95% CI 0.517-0.913) in the overall population and 0.613 (95% CI 0.434-0.843) amongst the emergency referrals. CONCLUSION: Physicians at local hospitals within a centralized system of delivering paediatric intensive care were able to maintain adequate assessment skills in recognition and requesting for appropriate transfers of the most ill and efficiently utilized resources available at the regional centre. PMID- 11398702 TI - Methodological investigation of measuring nasopharyngeal temperature as noninvasive brain temperature analogue in the neonate. AB - OBJECTIVES: (a) To investigate in a newborn animal model whether nasopharyngeal temperature is more closely related to epidural brain temperature than rectal temperature and (b) to investigate in human neonates whether measurement of nasopharyngeal temperature is dependent on the measurement site and other conditions. DESIGN AND SETTING: (a) Animal experiment in newborn piglets, at an institute for surgical research. (b) Prospective study in human neonates, at a neonatal intensive care unit of a tertiary care university hospital. ANIMALS AND PATIENTS: (a) Nineteen tracheostomized ventilated newborn piglets. (b) Twenty-two spontaneously breathing human newborns nursed either in an incubator or a cot. MEASUREMENTS AND RESULTS: (a) In the piglets nasopharyngeal temperature (Tnasoph) measured at the nose-ear distance, defined as distance from the inner brim of the nostril to the tragus and inner rim of the meatus accusticus, most closely reflected epidural temperature (Tepidur) at the epidural surface (r2 = 0.89), followed by skin temperature at the temple, rectal temperature (Trectum) at 2 cm depth, and esophageal temperature (Tesoph) in the middle esophagus. Tnasoph did not significantly differ before and after tracheostomy. (b) In the newborns Tnasoph was significantly lower than Trectum. Measurements of Tnasoph at nose-ear distance within a feeding tube had a high precision and were unaffected by breathing or head turning. A nasopharyngeal probe was imaged by magnetic resonance imaging in four newborns of various body weight; its tip when inserted to a depth equal to nose-ear distance was anatomically closest to the brain base but separated from it by tissue layer 2.2 cm thick. CONCLUSIONS: Tnasoph measured at a position anatomically closest to the brain reflects epidural brain temperature more closely than Trectum. When measured at nose-ear distance it is unaffected by breathing or head turning. Measuring Tnasoph within a feeding tube and standardizing the measuring position is crucial for its use as brain temperature analogue. PMID- 11398703 TI - Validity of applying TRISS analysis to paediatric blunt trauma patients managed in a French paediatric level I trauma centre. AB - OBJECTIVE: Using a weighted combination of the Revised Trauma Score (RTS), the Injury Severity Score (ISS), the type of injury (blunt or penetrating) and patient age, the TRISS method is used to calculate the probability of survival (ps) in trauma patients. The goal of this study was to compare the ability of the American Major Trauma Outcome Study (MTOS) norm for adult blunt trauma patients (ADULT) and the specific norm for paediatric patients (PED) to estimate the ps of injured children using TRISS methodology. DESIGN: Retrospective analysis using a paediatric trauma patient database. SETTING: A French level 1 paediatric trauma centre. PATIENTS: Four hundred seven consecutive paediatric blunt trauma patients, treated over a 3-year period. MEASUREMENTS: The observed and expected survivals were compared, using the M, W and Z scores, with both ADULT and PED. The W score is the number of survivors more or less than expected from the MTOS predictions for 100 patients. A Z score, which measures the significance of W, between -1.96 and +1.96, indicates no significant difference between observed and expected survivors. A value of M less than 0.88 indicates a disparity in the severity match between the study group and the MTOS group. We calculated the standardised W score (Ws), which represents the W score that would have been observed if the case mix of severity was identical to that of the MTOS group. Accordingly, a standardised Z score (Zs) was also calculated. In addition, we calculated the area under the receiver operating curve (aROC) using both norms, while calibration was also assessed by calculation of the Hosmer-Lemeshow goodness-of-fit tests. RESULTS: Using PED, the number of actual survivors (n = 364) was not significantly different from the MTOS (n = 358). The value of M, 0.65, indicated a disparity in the severity match between the study group and the MTOS group, due to a higher proportion of patients with lower ps (TRISS < 0.95, 52 vs 27%). We was +1.06% (95% confidence interval -0.34 to 2.08) and Zs was 1.48, indicating no significant difference from the MTOS. Using ADULT, the number of observed survivors (n = 364) was significantly higher than that expected (n = 354), with a W score of +2.70% (Z = +1.98, p < 0.05). There was a disparity in the severity match (M = 0.67) between the study group and the MTOS group, due to a higher proportion of patients with lower ps. Ws was +1.32% (95% confidence interval -0.12 to 2.37) and Zs = +1.79 (NS), indicating no significant difference from the MTOS. The Hosmer-Lemeshow statistics indicated that ADULT (Cg = 7.24, p = 0.51; Hg = 4.45, p = 0.81) and PED (Cg = 6.08, p = 0.64; Hg = 3.55, p = 0.90) provided sufficient goodness-of-fit. There was no significant difference in the aROC of the TRISS between the two norms (0.935 +/- 0.050 vs 0.936 +/- 0.050; NS). CONCLUSION: Both adult and paediatric norms were equally good predictors of the probability of survival of injured children, provided that Ws and Zs are used when there is a disparity in the severity match between the study group and the MTOS group. PMID- 11398704 TI - Dexamethasone inhibits stimulation of pulmonary endothelins by proinflammatory cytokines: possible involvement of a nuclear factor kappa B dependent mechanism. AB - OBJECTIVES: Recent studies have indicated effectiveness of glucocorticoid (GC) treatment in late, fibroproliferative adult respiratory distress syndrome. There is furthermore growing evidence for a role of endothelin-1 (ET-1) in lung fibroproliferation, but the impact of GC on stimulated pulmonary ET-1 is not well defined. DESIGN AND SETTING: Prospective study in an experimental laboratory. SUBJECTS: Male Wistar rats. INTERVENTIONS: Isolated lungs were perfused over 120 min in recirculatory mode in presence of vehicle, interleukin-1 beta (IL-1 beta; 5 ng/ml) plus tumor necrosis factor-alpha (TNF-alpha; 5 ng/ml), dexamethasone (Dx; 1 nmol/l), Dx (10 nmol/l), IL-1 beta plus TNF alpha plus Dx 1, or IL-1 beta plus TNF alpha plus Dx 10 (n = 6 each). Pulmonary artery endothelial cells were stimulated over 30 min using a similar protocol. MEASUREMENTS AND RESULTS: Control lungs released 15.2 +/- 0.6 pg/ml big ET-1 and 0.46 +/- 0.06 pg/ml ET-1, and contained 0.73 +/- 0.05 ng/g wet weight (ww) big ET-1 and 3.06 +/- 0.22 ng/g ww ET-1. IL-1 beta plus TNF-alpha increased release of big ET-1 and ET-1, to 220% and 217%, and lung content of peptides, to 236% and 230%. Dx dose-dependently inhibited the cytokine-induced rise in peptide release and lung content and completely suppressed these effects at 10 nmol/l. Electrophoretic mobility shift assays with nuclear extracts of pulmonary artery endothelial cells demonstrated nuclear binding of the transcription factor nuclear factor kappa B in response to IL-1 beta plus TNF-alpha, which was decreased in presence of Dx 1 and Dx 10. CONCLUSIONS: GC inhibit the cytokine-induced upregulation of pulmonary vascular and tissue endothelins, possibly via nuclear factor kappa B dependent mechanisms. This finding may reinforce the therapeutic employment of GC in late ARDS. PMID- 11398705 TI - Increased ileal-mucosal-arterial PCO2 gap is associated with impaired villus microcirculation in endotoxic pigs. AB - OBJECTIVE: To investigate whether an increased ileal-mucosal-arterial PCO2 gap (delta PCO2) during hyperdynamic porcine endotoxemia is associated with impaired villus microcirculation. DESIGN: Prospective, randomized, controlled, experimental study. SETTING: Animal research laboratory. ANIMALS: Twenty-two domestic pigs. INTERVENTIONS: After baseline measurements, anesthetized and ventilated pigs received continuous i.v. endotoxin (ETX, n = 12) for 24 h or placebo (SHAM, n = 10). MEASUREMENTS AND RESULTS: Before, as well as 12 and 24 h after, the start of endotoxin or saline portal venous blood flow (QPV, ultrasound flow probe) and lactate/pyruvate ratios (L/P), the ileal-mucosal-arterial delta PCO2 (fiberoptic sensor) and bowel-wall capillary hemoglobin O2 saturation (%Hb O2-cap, remission spectrophotometry) were assessed together with intravital video records of the ileal-mucosal microcirculation (number of perfused/heterogeneously perfused/unperfused villi) using orthogonal polarization spectral imaging (CYTOSCAN A/R) via an ileostomy. At 12 and 24 h endotoxin infusion, about half of the evaluated villi were heterogeneously or unperfused which was paralleled by a progressive significant increase of the ileal-mucosal-arterial delta PCO2 and portal venous L/P ratios, whereas QPV as well as both the mean %Hb-O2-cap and the %Hb-O2-cap frequency distributions remained unchanged. By contrast, in the SHAM group, mucosal microcirculation was well-preserved, and none of the other parameters were influenced. CONCLUSIONS: We conclude that an increased ileal mucosal-arterial delta PCO2 during porcine endotoxemia is related to impaired villus microcirculation. A putative contribution of disturbed cellular oxygen utilization resulting from "cytopathic hypoxia" may also assume importance. PMID- 11398706 TI - Determination of total blood volume by indicator dilution: a comparison of mean transit time and mass conservation principle. AB - OBJECTIVE: Using indocyanine green (ICG), blood volume can be determined within minutes according to the mass conservation principle by back-extrapolation of the concentration/time curve to the time of injection (BVTinj) or by the transit time approach (BVMTT) as the product of cardiac output and mean transit time (MTT) of ICG through the circulation. To see which factor accounts for the difference between the two methods we measured cardiac output and MTT independently and compared the volumes with those obtained by dilution of Evans blue (BVEB). DESIGN: Prospective animal study. SETTINGS: University department of experimental anaesthesiology. ANIMALS: Six anaesthetised, spontaneously breathing dogs with chronically implanted ultrasound flow probes around the pulmonary artery. MEASUREMENTS AND RESULTS: BVMTT and BVTinj agreed closely (48 +/- 2 ml.kg-1 and 49 +/- 2 ml.kg-1), but underestimated blood volume by about 40% compared with BVEB (75 +/- 1 ml.kg-1). Transit times measured were 33 +/- 1 s and should be about 50 s as calculated from the quotient of BVEB and cardiac output. CONCLUSIONS: Both methods underestimate blood volume by about the same extent compared with BVEB, probably because slowly perfused compartments are not detected during the short measurement period of 4 min. In the case of the transit time approach, rather short transit times result and in the case of the mass conservation principle, back-extra-polation yields rather high plasma concentrations of ICG at the time of injection. Accordingly, the two methods seem to be equivalent for measuring blood volume rapidly, although the absolute volume is underestimated by about 40%. PMID- 11398707 TI - Effect of an acute increase in PCO2 on splanchnic perfusion and metabolism. AB - OBJECTIVES: To evaluate the acute effects of an increase in PCO2 on splanchnic tissue perfusion and metabolism. DESIGN: Clinical prospective study in the intensive care unit in a university clinic. PATIENTS: Six patients with severe acute lung injury requiring mechanical ventilation. All patients had bilateral infiltrates on chest radiography, a PaO2/FIO2 ratio less than 200 mmHg, and stable hemodynamics without vasoactive drugs. INTERVENTIONS: PCO2 was increased 5 20% by an added dead space from baseline, followed by a return to baseline. MEASUREMENTS AND RESULTS: Splanchnic blood flow was measured using primed continuous infusion of indocyanine green dye with hepatic venous sampling and systemic hemodynamics by routine monitoring; we also determined the gastric mucosal-arterial PCO2 difference and splanchnic lactate/pyruvate exchange. PCO2 was increased by an added dead space; after 60 min all measurements were repeated; after return to baseline a third measurement followed. The increase in PCO2 had no significant effect on splanchnic blood flow or indices of perfusion and metabolism. CONCLUSION: Our results suggest that acute moderate changes in PCO2 have no major effect on splanchnic perfusion and metabolism. PMID- 11398708 TI - Outcome of long-stay intensive care patients. AB - OBJECTIVES: To assess the numbers, characteristics and outcome for patients requiring long-term intensive care. DESIGN AND SETTING: Observational cohort study in 23 Scottish intensive care units over a 3-year period. PATIENTS AND PARTICIPANTS: 323 patients with an ICU stay of 30 days or more. MEASUREMENTS AND RESULTS: Although representing only 1.6% of patients, those with long stays occupied 15.7% of bed-days. Hospital survival among these patients was 59.9%. With the available data it was not possible to discriminate survivors from non survivors. CONCLUSIONS: Since these patients have a relatively high hospital survival, resources should not be withheld from them on the basis of prolonged ICU stay alone, even in countries with limited ICU provision. PMID- 11398709 TI - Delayed complication of central venous catheterisation after prone positioning. PMID- 11398710 TI - Rapid diagnosis of alcoholic ketoacidosis by proton NMR. AB - In alcoholic patients, metabolic acidosis can be related to lactate acidosis associated with sepsis or thiamine deficiency, ketoacidosis, methanol or ethylene glycol poisoning. High resolution proton nuclear magnetic resonance (NMR) can be used to detect abnormal organic acid metabolites in urine or serum from patients with various metabolic disorders. In the present case, a 26-year-old patient was admitted for a coma associated with severe metabolic acidosis. Alcoholic ketoacidosis (AKA) was identified by urine proton NMR. Her metabolic disorders rapidly improved. Persisting associated neurological alteration was related to extrapontine myelinolysis as shown by imaging cerebral NMR. PMID- 11398711 TI - New evidence for old therapies in catecholamine-dependent septic shock. PMID- 11398712 TI - Prevalence of infections and systemic inflammatory response syndrome in southern Spanish ICUs. PMID- 11398713 TI - Acute respiratory distress syndrome associated with accidental hypothermia. PMID- 11398714 TI - Lesson of the month: tracheal perforation in an infant after tracheal surgery. PMID- 11398716 TI - Case report: patient's perspective on acute diabetic neuropathy. PMID- 11398715 TI - Diabetic foot ulcers. Pathophysiology, assessment, and therapy. AB - OBJECTIVE: To review underlying causes of diabetic foot ulceration, provide a practical assessment of patients at risk, and outline an evidence-based approach to therapy for diabetic patients with foot ulcers. QUALITY OF EVIDENCE: A MEDLINE search was conducted for the period from 1979 to 1999 for articles relating to diabetic foot ulcers. Most studies found were case series or small controlled trials. MAIN MESSAGE: Foot ulcers in diabetic patients are common and frequently lead to lower limb amputation unless a prompt, rational, multidisciplinary approach to therapy is taken. Factors that affect development and healing of diabetic patients' foot ulcers include the degree of metabolic control, the presence of ischemia or infection, and continuing trauma to feet from excessive plantar pressure or poorly fitting shoes. Appropriate wound care for diabetic patients addresses these issues and provides optimal local ulcer therapy with debridement of necrotic tissue and provision of a moist wound-healing environment. Therapies that have no known therapeutic value, such as foot soaking and topical antiseptics, can actually be harmful and should be avoided. CONCLUSION: Family physicians are often primary medical contacts for patients with diabetes. Patients should be screened regularly for diabetic foot complications, and preventive measures should be initiated for those at risk of ulceration. PMID- 11398717 TI - Rages and refusals. Managing the many faces of adolescent anxiety. AB - OBJECTIVE: To provide family physicians with a practical approach to recognition, assessment, and treatment of adolescent anxiety disorders complicated by avoidance or oppositional behaviour. QUALITY OF EVIDENCE: Current literature was searched via MEDLINE using the MeSH headings Anxiety and Anxiety Disorders, focusing on epidemiology, clinical presentations in adolescence, and both pharmacologic and nonpharmacologic treatment. In addition, internationally accepted diagnostic criteria, current practice guidelines, and recent textbooks by recognized experts were reviewed. Research evidence and consensus recommendations were integrated with a practical approach developed in a provincial mood and anxiety disorders clinic. MAIN FINDINGS: Anxiety disorders are common in adolescents, with estimated prevalence of at least 10%. Substance abuse and avoiding school are common complications, and irritability with behavioural and rage problems can interfere with effective management. Current controlled research is examining the effectiveness of serotonergic medications known to benefit panic disorder, social phobia, and generalized anxiety in adults. While cognitive and behavioural treatments are effective for some child and adolescent anxiety disorders, they can be difficult to administer, and a supportive and psychoeducational approach could be as effective for those who refuse to go to school. CONCLUSION: Family physicians' awareness of the role of anxiety in adolescent school avoidance and in intense, oppositional emotional reactions at home can lead to more specific assessment and therapeutic intervention. Practical management strategies are recommended. PMID- 11398718 TI - Early Canadian surgery. A shorebird's-eye view. PMID- 11398719 TI - Don't just look; touch! PMID- 11398720 TI - Hypothesis: the research page. Part 2: Confidence intervals and P values. PMID- 11398721 TI - Hospital care by family physicians. Exodus or opportunity? PMID- 11398722 TI - Patents in therapeutics. A divisive influence. PMID- 11398723 TI - Hats off to family physicians. PMID- 11398724 TI - Himalayan Institute Hospital and medical city. A new model of health care and education. PMID- 11398725 TI - Taking drugs during pregnancy. How safe are the unsafe? AB - QUESTION: I prescribed misoprostol to one of my patients with a peptic ulcer. When she found out she was pregnant while on the drug, both she and, admittedly, I were very scared to learn that the drug is teratogenic in that it causes Mobius syndrome. How great is the risk? ANSWER: Very small. Although women who use misoprostol during the first trimester have a 30-fold higher risk of having babies with Mobius syndrome, the malformation is so rare that, even if you see 1000 women who took misoprostol during embryogenesis, you might not see a single child with the syndrome. It is crucial to explain the size of the risk; otherwise women tend to believe the risk is huge even when, in fact, it is hardly measurable. PMID- 11398726 TI - Ophthaproblem. Photodynamic therapy with verteporfin. PMID- 11398727 TI - Release of medical records. PMID- 11398728 TI - Clinical diagnosis of acute sinusitis in adults. PMID- 11398729 TI - Do hip pads help prevent hip fractures? PMID- 11398730 TI - Role of family physicians in hospitals. Did it change between 1977 and 1997? AB - OBJECTIVE: To investigate whether hospital activities and attitudes toward hospitals of members of an urban family medicine department changed between 1977 and 1997. To explore whether these activities and attitudes are different among fee-for-service (FFS) and non-FFS physicians in 1997. DESIGN: Cross-sectional surveys by interview (1977) and self-administered questionnaire (1997). SETTING: Community-based family practices in Hamilton, Ont. PARTICIPANTS: In 1977, 88 of 89 (98.9%) and, in 1997, 66 of 88 (75.0%) members of the Department of Family Medicine at St Joseph's Hospital in Hamilton. MAIN OUTCOME MEASURES: Perceived reasons for involvement in hospital work; time spent and main activities in hospital; use of hospital privileges; attitudes toward family physicians' role in hospital, hospital work, and the Department of Family Medicine; perceptions of patients', consultants', and hospital administrators' attitudes toward family physicians' role in hospitals. RESULTS: In 1977 and 1997, patient care and continuing education remained key reasons for doing hospital work. In 1997, however, respondents spent a mean of 3 hours less per week in hospital; used the hospital less often for procedures, meetings, and teaching; and assumed less responsibility for their patients' in-hospital care. While perceptions of hospital work changed over the years, most physicians continued to see a need and have a desire to remain involved in hospitals. Fee-for-service and non-FFS physicians held different opinions on the needs of both hospitalized patients and family physicians. CONCLUSION: Although physicians' hospital activities and attitudes changed between 1997 and 1997, most continued to see a need and have a desire to remain involved in hospitals. PMID- 11398731 TI - Short report: does training in a family practice inpatient service affect practice after graduation? PMID- 11398732 TI - [Screening for violence against women. Validation and feasibility studies of a French screening tool]. AB - OBJECTIVE: To replicate, in a Francophone community, our prior work determining the reliability and validity of the full Woman Abuse Screening Tool (WAST) and a two-item version (WAST-Short). DESIGN: Questionnaires completed by abused and nonabused women. SETTING: Two women's shelters in Francophone communities in Ontario and Quebec and participants' homes or workplaces. PARTICIPANTS: A convenience sample of 25 abused women currently residing in two women's shelters and a convenience sample of 21 women who reported they were not abused. MAIN OUTCOME MEASURES: Women's responses to French versions of the WAST, the Abuse Risk Inventory (ARI), and comfort in answering the questions were compared. Also, the reliability and validity of French versions of WAST and WAST-Short were assessed. RESULTS: Abused (n = 23) and not abused (n = 21) women were demographically similar. A strong single-factor structure that accounted for 81% of total variance in the French WAST items was identified. The French WAST was found to be highly reliable with a coefficient alpha of .95 and demonstrated construct and discriminant validity. The WAST-Short correctly classified all the nonabused women and 78.7% of the abused women. The abused women reported feeling less comfortable responding to the WAST questions than the nonabused women. CONCLUSION: The French version of the WAST demonstrated good reliability and validity and discriminated between known samples of abused and nonabused women. Even though the French WAST-Short did not perform as well as the English version, results of this study support further evaluation of the WAST for screening women in Francophone or bilingual family practice settings. PMID- 11398733 TI - Do residents in a northern program have better quality lives than their counterparts in a city? AB - OBJECTIVE: To determine whether McMaster University's family medicine residents training in the Family Medicine North (FMN) program have better quality lives than those based in Hamilton, Ont (urban). DESIGN: Residents at both sites were simultaneously given the Quality of Life Questionnaire, a standardized measurement tool. They were asked to complete the questionnaire anonymously and to provide demographic data. SETTING: Family practice residencies in Ontario. PARTICIPANTS: McMaster University's family medicine residents. Of 66 residents living in Hamilton, 36 completed the questionnaire; five respondents were ineligible. Of 25 residents living in Thunder Bay, Ont, 24 completed the questionnaire; none were ineligible. MAIN OUTCOME MEASURES: Total quality-of-life score. Score was divided into five major domains, each with several subdomains: general well-being (material, physical, and personal growth), interpersonal relations (marital, parent-child, extended family, and extramarital), organizational activity (altruistic and political behaviour), occupational activity (job characteristics, occupational relations, and job satisfiers), and leisure and recreational activity (creative/esthetic behaviour, sports activity, vacation behaviour). RESULTS: The FMN residents scored significantly higher than the Hamilton-based residents on overall quality of life (124.7 vs 112.5, P < .05) and tended to score higher in the five major domains. The trend reached statistical significance in general well-being and occupational activity; it was also apparent in various subdomains, with statistically significant differences in material well-being, marital relations, job characteristics, job satisfiers, and vacation behaviour. CONCLUSION: Family Medicine North residents enjoy better quality of life than their urban counterparts based on responses to a standardized questionnaire. PMID- 11398734 TI - Clearing the air. Stopping infection through better IAQ control. PMID- 11398735 TI - Managing your 'energy assets'. Here's a step-by-step approach to getting started. PMID- 11398736 TI - Nurturing design. A look at recommended newborn ICU standards. PMID- 11398737 TI - Clean enough to eat from. Sanitizing and maintaining food preparation and service areas. PMID- 11398738 TI - The Medicare Anti-Kickback statute: understanding the intent requirement. PMID- 11398739 TI - Workplace violence: practical advice for a problem with dire legal consequences. PMID- 11398740 TI - Group practice structure and compensation plans under Stark II regulations. PMID- 11398741 TI - Combination antiretroviral therapy and duration of pregnancy. AB - OBJECTIVE: To assess the association between type and timing of initiation of antiretroviral therapy in pregnancy and duration of pregnancy. DESIGN: Prospective study. METHODS: Data on 3920 mother-child pairs were examined (3015 mother-child pairs from the European Collaborative Study and 905 from the Swiss Mother + Child HIV Cohort Study). Factors examined included gestational age, antiretroviral therapy during pregnancy, maternal CD4 count, viral load, illicit drug use (IDU) and mode of delivery. Deliveries at less than 37 weeks were defined as premature. RESULTS: The prematurity rate was 17% and median gestational age 39 weeks. Twenty-three per cent (896 of 3920) of women received antiretroviral therapy during pregnancy: 64% (573 of 896) zidovudine monotherapy, 24% (215) combination therapy without protease inhibitors (PI) and 12% (108) combination therapy with PI. In multivariate analysis, adjusted for maternal CD4 count and IDU, odds ratio (OR) of prematurity was 2.60 [95% confidence interval (CI), 1.43-4.75] and 1.82 (95% CI, 1.13-2.92) for infants exposed to combination therapy with and without a PI, respectively, compared to no treatment. Exposure to monotherapy was not associated with prematurity, but severe immunosuppression and IDU in pregnancy were. Women on combination therapy from before pregnancy were twice as likely to deliver prematurely as those starting therapy in the third trimester (OR, 2.17; 95% CI, 1.03-4.58). CONCLUSIONS: Pregnancy issues should be discussed when making decisions about initiation of combination antiretroviral therapy for HIV-infected women. Elective caesarean section to reduce vertical transmission at 36 weeks rather than 38 weeks may be advisable in women on combination therapy with PI. PMID- 11398742 TI - Transumbilical breast augmentation: a new instrument for changing implants from the subglandular to the subpectoral position. PMID- 11398743 TI - Bronchioloalveolar carcinoma: clinical, radiographic, and pathological findings. Surgical results. AB - BACKGROUND: Bronchioloalveolar carcinoma (BAC) remains one of the most controversial of lung cancer subtypes. METHODS: From 1980 to 1998, 374 resected patients for NSCLC were followed up in our department. Of the 147 cases histologically defined as adenocarcinoma, 34 were pure BAC. The records of these 34 patients were retrospectively reviewed in order to evaluate patient and tumor characteristics and to identify which variables had a prognostic impact on survival and recurrence rate. RESULTS: Patient age, sex, smoking habits and symptoms were not differentiating characteristics when related to radiographic presentation or to natural history. Mucinous cell-type (23.6% of cases) was more frequent with non-smokers, presence of a single nodule or mass and stage I. Favorable characteristics were: a) the prevalence of stage I and N0 cases (59% and 76.7% of cases, respectively) with a mean survival time of 66 and 77 months, respectively; and b) the radiographic presentation of a solitary pulmonary nodule or mass (76.4% of cases), that, independently of nodal involvement, showed a higher mean survival time (62 months). Independently significant adverse prognostic factors were: stage II-IV, lymph node involvement, and patient age over sixty years. The radiographic presence of multiple or satellite nodules was related to a significantly adverse prognosis (mean survival time: 18 months) by univariate analysis; this was not confirmed by multivariate analysis. CONCLUSIONS: In our experience BAC was the NSCLC subtype more frequently associated with a good outcome after resection; surgery should not to be denied also in patients with multiple nodules, when under sixty years of age and no lymph node involvement. PMID- 11398744 TI - Specific endocytosis and catabolism in the scavenger endothelial cells of cod (Gadus morhua L.) generate high-energy metabolites. AB - The catabolic fate of circulating hyaluronan and the proteoglycan chondroitin sulphate (CSPG) was studied in the Atlantic cod (Gadus morhua L.). Distribution studies using radio-iodinated ligand demonstrated that CSPG was rapidly eliminated from the blood by the endocardial endothelial cells (EECs) of the heart atrium and ventricle. The presence of excess amounts of hyaluronan or CSPG inhibited uptake of [125I]hyaluronan into cultured atrial EECs (aEECs) by 46% and 84%, respectively. Neither formaldehyde-treated serum albumin (FSA) nor mannose inhibited this uptake. The presence of excess amounts of CSPG and hyaluronan inhibited uptake of [125I]CSPG by 90% and 42%, respectively, suggesting that aEECs express a specific hyaluronan binding site that also recognizes CSPG. FSA inhibited endocytosis of [1251]CSPG by 65%, indicating that CSPG is also recognized by the scavenger receptor. Approximately 17% and 57% of added [125I]hyaluronan and 15% and 65% of the added [125I]CSPG were endocytosed after 1 and 24h, respectively. High-performance liquid chromatographic analyses of the spent medium after endocytosis of hyaluronan and CSPG serglycin labelled biosynthetically with 3H in the acetyl groups identified labelled the low molecular-mass degradation products as [3H]acetate, indicating that aEECs operate anaerobically. These findings suggest that acetate released from cod EECs following catabolism of endocytosed hyaluronan and CSPG represents a high-energy metabolite that may fuel cardiomyocytes. PMID- 11398745 TI - Hydrodynamic drag of diving birds: effects of body size, body shape and feathers at steady speeds. AB - For birds diving to depths where pressure has mostly reduced the buoyancy of air spaces, hydrodynamic drag is the main mechanical cost of steady swimming. Drag is strongly affected by body size and shape, so such differences among species should affect energy costs. Because flow around the body is complicated by the roughness and vibration of feathers, feathers must be considered in evaluating the effects of size and shape on drag. We investigated the effects of size, shape and feathers on the drag of avian divers ranging from wing-propelled auklets weighing 75 g to foot-propelled eiders weighing up to 2060 g. Laser scanning of body surfaces yielded digitized shapes that were averaged over several specimens per species and then used by a milling machine to cut foam models. These models were fitted with casts of the bill area, and their drag was compared with that of frozen specimens. Because of the roughness and vibration of the feathers, the drag of the frozen birds was 2-6 times that of the models. Plots of drag coefficient (C(D)) versus Reynolds number (Re) differed between the model and the frozen birds, with the pattern of difference varying with body shape. Thus, the drag of cast models or similar featherless shapes can differ both quantitatively and qualitatively from that of real birds. On the basis of a new towing method with no posts or stings that alter flow or angles of attack, the dimensionless C(D)/Re curves differed among a size gradient of five auklet species (75-100g) with similar shapes. Thus, extrapolation of C(D)/Re curves among related species must be performed with caution. At lower speeds, the C(D) at a given Re was generally higher for long-necked birds that swim with their neck extended (cormorants, grebes, some ducks) than for birds that swim with their head retracted (penguins, alcids), but this trend was reversed at high speeds. Because swimming birds actually travel at a range of instantaneous speeds during oscillatory strokes, species variations in drag at different speeds must be considered in the context of accelerational stroking. PMID- 11398746 TI - The influence of stimulus and background colour on signal visibility in the lizard Anolis cristatellus. AB - Anoline lizards communicate with visual displays in which they open and close a colourful throat fan called the dewlap. We used a visual fixation reflex as an assay to test the effects of stimulus versus background chromatic and brightness contrast on the probability of detecting a moving coloured (i.e. dewlap-like) stimulus in Anolis cristatellus. The probability of stimulus detection depended on two additive visual-system channels, one responding to brightness contrast and one responding to chromatic contrast, independent of brightness. The brightness channel was influenced only by wavelengths longer than 450nm and probably received input only from middle- and/or long-wavelength photoreceptors. The chromatic contrast channel appeared to receive input from three, or possibly four, different classes of cone in the anoline retina, including one with peak sensitivity in the ultraviolet. We developed a multi-linear regression equation that described most of the results of this study to a reasonable degree of accuracy. In the future, this equation could be used to predict the relative visibility of different-coloured stimuli in different habitat light conditions, which should be very useful for testing hypotheses that attempt to relate habitat light conditions and visual-system response to the evolution of signal design. PMID- 11398747 TI - Ventilatory modes and mechanics of the hedgehog skate (Leucoraja erinacea): testing the continuous flow model. AB - The movement of water across the gills of non-ram-ventilating fishes involves the action of two pumps: a pressure pump that pushes water across the gills from the oropharyngeal to the parabranchial cavity, and a suction pump that draws water across the gills from the oropharyngeal into the parabranchial cavity. Together, the two are thought to keep water flowing continuously anteroposteriorly through the head of the respiring animal. However, there is evidence that the pressure and suction pumps do not always work in perfect phase in elasmobranch fishes, leading to periods of higher pressure in the parabranchial than in the oropharyngeal cavity. We investigated the existence and consequence of such pressure reversals in the hedgehog skate Leucoraja erinacea using pressure transducers, sonomicrometry and flow visualization including internal visualization using endoscopy. We noted four patterns of respiration in the experimental skates distinguished by the flow pattern at the three openings into the respiratory system: (1) in through the spiracle only, (2) in through the mouth + spiracle, (3) in through the mouth only, and (4) the mouth held open throughout the respiratory cycle. The first two were by far the dominant modes recorded from experimental animals. We determined that pressure reversals exist in the hedgehog skate, and that the gill bars adducted during such pressure reversals. Direct observation confirmed that these pressure reversals do correspond to pulsatile flow across the gills. During mouth+spiracle ventilation the flow completely reversed direction, flowing from the parabranchial chambers back across the gills and into the oropharyngeal cavity. Finally, we addressed the utility of sonomicrometry as a technique for determining kinematics in aquatic animals. Despite some problems involving errors inherent to the system design, we found the technique useful for complementing such techniques as pressure measurements and endoscopy. PMID- 11398748 TI - Stereotypic leg searching movements in the stick insect: kinematic analysis, behavioural context and simulation. AB - Insects are capable of efficient locomotion in a spatially complex environment, such as walking on a forest floor or climbing in a bush. One behavioural mechanism underlying such adaptability is the searching movement that occurs after loss of ground contact. Here, the kinematic sequence of leg searching movements of the stick insect Carausius morosus is analysed. Searching movements are shown to be stereotypic rhythmic movement sequences consisting of several loops. The typical loop structure allows the mean tarsus trajectory to be calculated using a feature-based averaging procedure. Thus, it is possible to describe the common underlying structure of this movement pattern. Phase relationships between joint angles, analysed for searching front legs, indicate a central role for the thorax-coxa joint in searching movements. Accordingly, the stereotyped loop structure of searching differs between front-, middle- and hindlegs, with leg-specific patterns being caused by differing protraction/retraction movements in the thorax-coxa joint. A simple artificial neural network that had originally been devised to generate simple swing movements allows two essential features of empirical searching trajectories to be simulated: (i) cyclic movements and (ii) the smooth transition into a search trajectory as a non-terminated swing movement. It is possible to generate several loops of a middle-leg search, but the precise size and shape of the loops fall short of a real-life approximation. Incorporation of front-leg retraction or hind leg protraction during searching will also require an extension to the current model. Finally, front-leg searching occurs simultaneously with antennal movements. Also, because leg searching movements are a local behaviour, the legs remaining on the ground continue their stance phase, causing a forward shift of the body, including the searching leg. As a result of this shift, the centre of the searched space is close to the anterior extreme position of the tarsus during walking, representing the location of most likely ground contact according to past experience. Therefore, the behavioural relevance of searching movements arises from the combined actions of several limbs. PMID- 11398749 TI - Reduction in the rates of protein and amino acid catabolism to slow down the accumulation of endogenous ammonia: a strategy potentially adopted by mudskippers (Periophthalmodon schlosseri snd Boleophthalmus boddaerti) during aerial exposure in constant darkness. AB - This study was designed to elucidate the strategies adopted by mudskippers to handle endogenous ammonia during aerial exposure in constant darkness. Under these conditions, specimens exhibited minimal locomotory activity, and the ammonia and urea excretion rates in both Periophthalmodon schlosseri and Boleophthalmus boddaerti decreased significantly. As a consequence, ammonia accumulation occurred in the tissues of both species of mudskipper. A significant increase in urea levels was found in the liver of P. schlosseri after 24h of aerial exposure, but no similar increase was seen in the tissues of B. boddaerti. It is unlikely that these two species of mudskipper detoxified ammonia to urea during aerial exposure since B. boddaerti does not possess a complete ornithine urea cycle (OUC) and, although all the OUC enzymes were present in P. schlosseri, the activity of carbamoyl phosphate synthetase present in the liver mitochondria was too low to render the OUC functional for ammonia detoxification. Peritoneal injection of 15NH4Cl into P. schlosseri showed that this mudskipper was capable of incorporating some of the labelled ammonia into urea in its liver. However, aerial exposure did not affect this capability and did not induce detoxification of the accumulated ammonia to urea. Mudskippers exposed to terrestrial conditions and constant darkness did, however, show significant decreases in the total free amino acid content in the liver and blood, in the case of P. schlosseri and in the muscle of B. boddaerti. No changes in the alanine or glutamine content of the muscle were found in either species. Analyses of the balance between the reduction in nitrogenous excretion and the increase in nitrogenous accumulation further revealed that these two species of mudskipper were capable of reducing their protein and amino acid catabolic rates. Such adaptations constitute the most efficient way to avoid the build-up of internal ammonia, and would render unnecessary the detoxification of ammonia through energetically expensive pathways. This finding may be the first report of a teleost fish showing a reduction in proteolysis and amino acid catabolism in response to aerial exposure. PMID- 11398750 TI - Partial amino acid catabolism leading to the formation of alanine in Periophthalmodon schlosseri (mudskipper): a strategy that facilitates the use of amino acids as an energy source during locomotory activity on land. AB - When the mudskipper Periophthalmodon schlosseri was exposed to terrestrial conditions under a 12h:12h dark:light regime the fish could be very active, and levels of total free amino acids increased significantly in the muscle and plasma. Alanine levels increased threefold in the muscle, fourfold in the liver and twofold in the plasma. Similar phenomena were not observed in the more aquatic mudskipper, Boleophthalmus boddaerti. From these results, we concluded that P. schlosseri was capable of partial catabolism of certain amino acids to support activity on land. The amino groups of these amino acids were transferred directly or indirectly to pyruvate to form alanine. The resulting carbon chain was fed into the Krebs cycle and partially oxidized to malate, which could replenish pyruvate through the function of malic enzyme. This favourable ATP yield from partial amino acid catabolism was not accompanied by a net release of ammonia. Such an adaptation would be advantageous to P. schlosseri confronted with the problem of ammonia excretion during aerial exposure. Indeed, when P. schlosseri were forced to exercise on land after 24 h of aerial exposure, the alanine level in the muscles increased significantly, with no apparent change in glycogen content. In addition, there was no significant change in the ATP level and energy charge of the muscle. In contrast, when B. boddaerti were exercised on land, glycogen levels in the muscles decreased significantly and lactate levels increased. In addition, muscle energy charge was not maintained and the ATP level decreased significantly. Hence, it was concluded that when P. schlosseri were active on land, they were capable of using certain amino acids as a metabolic fuel, and avoided ammonia toxicity through partial amino acid catabolism. Such a strategy is the most cost-effective way of slowing down internal ammonia build-up without involving energy-expensive ammonia detoxification pathways. Furthermore, an examination of the balance between nitrogenous excretion and accumulation in a 70 g P. schlosseri revealed that degradation of amino acids in general was likely to be suppressed to slow down the build-up of ammonia internally. It is possible that such a strategy may be widely adopted, especially by obligatory air breathing fishes, to avoid ammonia intoxication during aerial exposure. PMID- 11398751 TI - Effects of serotonin, dopamine and ergometrine on locomotion in the pulmonate mollusc Helix lucorum. AB - The terrestrial snail Helix lucorum crawls using waves of muscular contraction (pedal waves) that spread along the sole of its foot. Crawling speed depends on wave generation frequency (step frequency) and the distance the snail moves forwards during each wave (step length). In a previous study, video recordings of a crawling snail showed that its sole length varied over a wide range and was directly correlated with mollusc speed. Speed depended on step length, which was directly related to sole length, rather than on step frequency, which remained rather constant. In the present study, the effects of dopamine, ergometrine (a blocker of dopamine receptors in molluscs) and serotonin injection on the linear relationship between sole length and locomotor speed in Helix lucorum were studied. In crawling snails, dopamine caused sole contraction, and locomotion slowed down or ceased. Ergometrine stimulated locomotion, which resembled rapid crawling with an extended sole, as observed under normal conditions. Serotonin stimulated locomotion and accelerated crawling significantly without causing changes in sole length. The acceleration of locomotion induced by serotonin injection was due to pedal wave (step) elongation. It is proposed that, during each locomotor episode, dopamine controls snail speed by regulating sole length, which determines the amplitude of contraction of the muscle cells involved in pedal waves and, as a result, step length; serotonin determines the basic step length and shifts the linear relationship between sole length and mollusc speed upwards along the axis of mollusc speed. The efficiency of the serotonergic system depends on the physiological state of the mollusc (e.g. that characteristic of summer or winter). PMID- 11398752 TI - The guidance of desert ants by extended landmarks. AB - Desert ants (Cataglyphis fortis) were trained to follow a fixed route around a barrier to a feeder. Their homeward trajectories were recorded on a test field containing a similar barrier, oriented either as in training or rotated through 22 or 45 . Under one set of experimental conditions, the homeward trajectories rotated with the orientation of the barrier, implying that the visual features of this extended landmark can determine the route independently of compass cues: the barrier provided a "visual scene" that controlled the trajectories of the ants. Under other conditions, the trajectories after rotation were a compromise between the habitual compass direction and the direction with respect to the rotated barrier. Trajectories were determined primarily by the visual scene when ants were allowed to return close to the nest before being caught and tested. The compromise trajectories were observed when ants were taken from the feeder. It seems that ants exhibit at least two separate learnt responses to the barrier: (i) a habitual compass direction triggered by the sight of the barrier and (ii) a visual scene direction that is compass-independent. We suggest that the weighting accorded to these different learnt responses changes with the state of the path integration system. PMID- 11398753 TI - Entraining the natural frequencies of running and breathing in guinea fowl (Numida meleagris). AB - Lung ventilation of tetrapods that synchronize their locomotory and ventilatory cycles during exercise could be economized if the resonant frequency of the respiratory system matched the animal's preferred step frequency. To test whether animals utilize this strategy, the input impedance of the respiratory system of five anesthetized, supine guinea fowl (Numida meleagris) was measured using a forced oscillation technique. The resonant frequency of the respiratory system was 7.12+/-0.27 Hz (N=5, mean +/- S.E.M.). No statistically significant difference was found between the resonant frequency of the respiratory system and the panting frequency used by guinea fowl at rest (6.67+/-0.16 Hz, N=11) or during treadmill locomotion (6.71+/-0.12 Hz, N=8) or to their preferred step frequency (6.73+/-0.09 Hz, N=7) (means +/- S.E.M.). These observations suggest (i) that, at rest and during exercise, panting guinea fowl maximize flow while expending minimal mechanical effort, and (ii) that natural selection has tuned the natural frequencies of the respiratory and locomotor systems to similar frequencies. PMID- 11398754 TI - Changes in the thoracic temperature of honeybees while receiving nectar from foragers collecting at different reward rates. AB - Mouth-to-mouth food exchange in eusocial insects (trophallaxis) contributes to the organization of complex social activities. In the case of honeybees, foragers returning from a nectar source transfer the food collected to receiver colony mates through oral contact. Previous studies have shown that the speed of nectar transfer within each contact (unloading rate) increases when foragers return from feeding sites with higher profitability, i.e. with more concentrated sugar solutions or higher solution flow rates. However, there is no evidence that the nectar unloading rate is actually evaluated by hive-mates during food exchange. To investigate this, trophallaxis between donor bees returning from a feeder with different flow rates of sucrose solution (range 1.0-8.2 microl min(-1) of 50% w/w sucrose solution) and receiver hive-mates was studied by combining behavioural and infrared thermal analysis. The results show that when foraging bees returned from a feeder delivering a higher flow rate they initiated unloading at higher thoracic temperatures and transferred the solution at higher speed. During these food exchanges, the thoraces of receiver bees warmed up faster in proportion to increasing forager temperature and unloading rate. Therefore, whatever the variable actually evaluated by receivers (mostly nectar processors, i.e. bees that handle nectar in the hive) during trophallaxis (unloading rate and/or donor thoracic temperature), they raised their activity level in proportion to that of the foragers. In this way, receiver bees will intensify their nectar processing when nectar foragers return from more profitable sites. PMID- 11398756 TI - Seasonal change in the capacity for supercooling by neonatal painted turtles. AB - Hatchlings of the North American painted turtle (Chrysemys picta) typically spend their first winter of life inside the shallow, subterranean nest where they completed incubation the preceding summer. This facet of their natural history commonly causes neonates in northerly populations to be exposed in mid-winter to ice and cold, which many animals survive by remaining unfrozen and supercooled. We measured the limit of supercooling in samples of turtles taken shortly after hatching and in other samples after 2 months of acclimation (or acclimatization) to a reduced temperature in the laboratory or field. Animals initially had only a limited capacity for supercooling, but they acquired an ability to undergo deeper supercooling during the course of acclimation. The gut of most turtles was packed with particles of soil and eggshell shortly after hatching, but not after acclimation. Thus, the relatively high limit of supercooling for turtles in the days immediately after hatching may have resulted from the ingestion of soil (and associated nucleating agents) by the animals as they were freeing themselves from their eggshell, whereas the relatively low limit of supercooling attained by acclimated turtles may have resulted from their purging their gut of its contents. Parallels may, therefore, exist between the natural-history strategy expressed by hatchling painted turtles and that expressed by numerous terrestrial arthropods that withstand the cold of winter by sustaining a state of supercooling. PMID- 11398755 TI - Rapid cold-hardening of Drosophila melanogaster (Diptera: Drosophiladae) during ecologically based thermoperiodic cycles. AB - In contrast to most studies of rapid cold-hardening, in which abrupt transfers to low temperatures are used to induce an acclimatory response, the primary objectives of this study were to determine (i) whether rapid cold-hardening was induced during the cooling phase of an ecologically based thermoperiod, (ii) whether the protection afforded was lost during warming or contributed to increased cold-tolerance during subsequent cycles and (iii) whether the major thermally inducible stress protein (Hsp70) or carbohydrate cryoprotectants contributed to the protection afforded by rapid cold-hardening. During the cooling phase of a single ecologically based thermoperiod, the tolerance of Drosophila melanogaster to 1 h at -7 degrees C increased from 5 +/- 5% survival to 62.5 +/- 7.3% (means +/- S.E.M., N=40-60), while their critical thermal minima (CTmin) decreased by 1.9 degrees C. Cold hardiness increased with the number of thermoperiods to which flies were exposed; i.e. flies exposed to six thermoperiods were more cold-tolerant than those exposed to two. Endogenous levels of Hsp70 and carbohydrate cryoprotectants were unchanged in rapidly cold hardened adults compared with controls held at a constant 23 degrees C. In nature, rapid cold-hardening probably affords subtle benefits during short-term cooling, such as allowing D. melanogaster to remain active at lower temperatures than they otherwise could. PMID- 11398757 TI - Modulation of intracellular transport of acidic fibroblast growth factor by mutations in the cytoplasmic receptor domain. AB - Endocytic uptake and intracellular transport of acidic fibroblast growth factor (aFGF) was studied in cells transfected with FGF receptor 4 with mutations in the cytoplasmic part. Endocytic uptake in HeLa cells was reduced but not abolished when the tyrosine kinase of the receptor was inactivated by mutations or deletions. The tyrosine kinase-dependent endocytosis of aFGF was prevented by the expression of a dominant negative dynamin mutant that blocks endocytosis from coated pits and caveolae. However, more than half of the total endocytic uptake of aFGF was not affected under these conditions, indicating an endocytic uptake mechanism not involving coated pits or caveolae. Mutation or deletion of a putative caveolin-binding sequence did not prevent the localization of part of the receptors to a low density, caveolin-containing subcellular fraction. Whereas wild-type receptor transfers the growth factor from early endosomes to the recycling compartment, kinase negative, full length receptors were inefficient in this respect and the growth factor instead accumulated in lysosomes. By contrast, when most of the intracellular part of the receptor, including the kinase domain, was removed, aFGF was transported to the recycling compartment, as in cells that express wild-type receptors, suggesting the presence of a kinase-regulated targeting signal in the cytoplasmic tail. PMID- 11398758 TI - Photo quiz. Eczema herpeticum. PMID- 11398759 TI - Gram-negative sepsis in neonates may be rising since start of GBS prophylaxis. PMID- 11398760 TI - [The "reversal sign." Apropos of a case]. PMID- 11398761 TI - [Depression in patient with neurological diseases. Proposed therapeutic strategies]. PMID- 11398762 TI - [Clinical study of patients with Alzheimer's disease under treatment with tacrine]. PMID- 11398763 TI - [Cerebral electrical tomography in congenital bilateral perisylvian syndrome]. PMID- 11398764 TI - [Adrenomyeloneuropathy. Apropos of a case]. PMID- 11398765 TI - Paroxysmal atrial fibrillation following marijuana intoxication: a two-case report of possible association. PMID- 11398766 TI - An asymptomatic giant pericardial cyst. PMID- 11398767 TI - Perinuclear-antineutrophil cytoplasmic antibodies are associated with vasculitis. AB - We describe a 62-year-old man with perinuclear-antineutrophil cytoplasmic antibodies-associated vasculitis, which involved the heart, lung, and kidneys. The patient's care was complicated by total occlusions of the brachiocephalic and right renal arteries and a stenosis of the left renal artery. Involvement of large-sized vessels has not been reported in patients with perinuclear antineutrophil cytoplasmic antibodies-associated vasculitis. PMID- 11398768 TI - Two cases of cardiac cyst hydatid with right and left ventricular involvement. AB - Cardiac cyst hydatic is a rare disease. Two cases with left and right ventricular involvement are presented that demonstrate the use of echocardiography in the diagnosis and during follow up of the disease. PMID- 11398769 TI - Recurrent obstructive mechanical valve thrombosis in mitral position. AB - We report a case of 37-year-old woman with recurrent prosthetic valve thrombosis (PVT). The patient was evaluated for hypercoagulable state and treated with streptokinase, tissue plasminogen activator and replacement of a bioprosthesis. PMID- 11398770 TI - Thromboangiitis obliterans a new look for an old disease. PMID- 11398771 TI - Severity of coronary artery disease and serum lipid levels. PMID- 11398772 TI - Coronary risk factors in patients with early-onset coronary disease and normal coronariography. PMID- 11398773 TI - Dye in the limbus fossa ovalis. PMID- 11398774 TI - My brilliant career--health promotion. Going public. Interview by Ann Dix. PMID- 11398775 TI - The 2000 Nobel Prize in Medicine or Physiology. PMID- 11398776 TI - Thoracic aortic aneurysm repair: improvement in operative results and its background. PMID- 11398777 TI - Effect of papaverine applications on blood flow of the internal mammary artery. AB - The aim of this prospective study was to compare the effect of different papaverine applications on the free blood flow in the internal mammary artery (IMA) prepared for coronary artery bypass grafting (CABG). The patients were divided into three groups: group I (n=50; intraluminal papaverine application), group II (n=50; topical papaverine application), and group III (n=50; periarterial papaverine application). The free flow from the distal cut end of the IMA was measured under controlled hemodynamic conditions (Flow 1). Just before the establishment of cardiopulmonary bypass, the flows of the IMA were measured again (Flow 2). The mean blood flow in the IMA was 60.7+/-6.0 ml/min before papaverine application. After papaverine application, the mean blood flow in group I was 129.3+/-10.0 ml/min, in group II, it was 87.7+/-3.8 ml/min, and in group III, it was 130.6+/-9.2 ml/min (p<0.0001). Proportional increases in blood flow observed in group I (106.3%) and group III (116.2%) were higher than in group II (44.2%) (p<0.0001). Consequently, to relieve perioperative spasm of the IMA, application of papaverine injected into the periarterial tissues of its pedicle was considered to be a safe and effective alternative to topical or intraluminal application. PMID- 11398778 TI - Advantages of rectus fascial slings for urinary incontinence in children with neuropathic bladders. AB - PURPOSE: Many surgical procedures to improve outlet resistance in children with neuropathic bladders are obstructive and increase the detrusor leak point pressure. In contrast, fascial slings are designed to achieve continence by increasing the Valsalva or stress leak point pressure without altering the detrusor leak point pressure. We evaluate the effectiveness of fascial slings in achieving continence in pediatric patients with neuropathic bladder. MATERIALS AND METHODS: From October 1994 until February 1999, 10 females and 8 males with neuropathic bladder secondary to myelodysplasia or traumatic spinal cord injury underwent fascial sling procedures. Mean patient age was 14 years (range 8 to 18) and all were incontinent despite aggressive medical management. Urodynamic evaluation was performed preoperatively and postoperatively. Specific urodynamic measurements included detrusor leak point pressure, stress leak point pressure and detrusor compliance. Compliance was only compared in the 12 nonaugmented cases. RESULTS: With a mean followup of 21.2 months (range 6 to 57), preoperative and postoperative urodynamics revealed little change in mean detrusor leak point pressure (23.2 versus 23.22 cm. H2O) but a substantial increase in mean stress leak point pressure (41.6 versus 64.5 cm. H2O). Mean compliance was unchanged in the nonaugmented group (22.00 versus 26.78 ml/cm. H2O). Four patients (22.22%) remained wet after surgery, of whom 2 were successfully treated with a repeat sling procedure and 1 with collagen injection for an overall continence rate of 94.44%. CONCLUSIONS: Fascial slings can be effectively used in pediatric patients for neuropathic incontinence. Furthermore, stress urinary incontinence is corrected by increasing the Valsalva or stress leak point pressure with preservation of the detrusor leak point pressure. Preservation of detrusor leak point pressure is particularly advantageous because other forms of bladder outlet procedures achieve continence at the expense of increasing detrusor pressures, thus placing the upper tracts at risk for damage. PMID- 11398779 TI - The impact of HIPAA electronic transmissions and health information privacy standards. PMID- 11398780 TI - APHA president-elect talks with JADA. Interview by Elaine R. Monsen. PMID- 11398781 TI - Interactions between HIV-related medications and methadone: an overview. Updated March 2001. PMID- 11398782 TI - Rationale for and the substantial potential benefits linked to early recognition and optimal treatment of psychotic disorders, specifically schizophrenia. PMID- 11398783 TI - Commentary: learning to love mistakes. PMID- 11398784 TI - Commentary: doctors are obliged to be honest with their patients. PMID- 11398785 TI - Commentary: a climate of secrecy undermines public trust. PMID- 11398786 TI - Responding rationally to recent report of abuse/diversion of Oxycontin. PMID- 11398787 TI - Medical education: a barrier to pain therapy and palliative care. PMID- 11398788 TI - Argentine palliative care standards. PMID- 11398789 TI - Setting up a "pain-free" hospital. PMID- 11398790 TI - Treatment of nausea and vomiting in long-term survivors of pancreatic cancer. PMID- 11398791 TI - Topiramate relieves idiopathic and symptomatic trigeminal neuralgia. PMID- 11398792 TI - Ketamine has equal affinity for NMDA receptors and the high-affinity state of the dopamine D2 receptor. PMID- 11398793 TI - Image of the month. Heriditary hemorrhagic telangiectasia. PMID- 11398794 TI - API2-MALT1 chimeric transcript is a predictive marker for the responsiveness of H. pylori eradication treatment in low-grade gastric MALT lymphoma. PMID- 11398795 TI - Why the mesenteric margin? PMID- 11398796 TI - Pathogenesis of diclofenac enteropathy. PMID- 11398797 TI - Organic anion-transporting polypeptides: expression cloning and functional tests. PMID- 11398798 TI - Practical. Loyalty dilemma. PMID- 11398799 TI - 1Q[3a]. Federal funding to rehab or replace rural facilities? PMID- 11398800 TI - Tech tackles errors. PMID- 11398801 TI - The Health Care 250. AB - Despite last year's dot-com meltdown, 2000 was a very good year for many health care companies. Pharmaceutical companies continue to fare well on Hospitals & Health Networks' annual compilation of the 250 largest health care companies. Those rehabilitation services companies that survived the shakeout also posted healthy gains. PMID- 11398802 TI - Mergers and acquisitions. PMID- 11398803 TI - Seventh Annual ASHE Vista Awards. Team buildings. PMID- 11398804 TI - Medication safety issue brief 5. Crucial role of therapeutic guidelines. AB - Clinical practice guidelines have gained favor as a key method for health care organizations to improve the quality and consistency of medical care. They can also be used to reduce the chance of error by establishing a standard protocol for how care is carried out. It sounds simple, but choosing and developing well designed guidelines can be challenging, as can persuading caregivers to accept a single way to do things. Still, guidelines based on solid evidence and good outcomes can make everyone's job easier. PMID- 11398805 TI - Creatures of culture. PMID- 11398806 TI - Immunologic approach to herpes simplex virus. AB - The fundamental biological characteristic of herpes simplex virus (HSV) infection is its ability to establish latency and periodically reactivate, resulting in productive infectious virus. Recurrent HSV infections occur in spite of the presence of host immune responses to the virus. Because genital herpes is currently one of the three most prevalent sexually transmitted diseases worldwide, its potential public health impact has contributed to increased awareness in medical communities. The current state of knowledge on the immunological control of primary and recurrent HSV infections is reviewed, as well as the various immune-based therapeutic approaches to resultant disease. Finally, the potential benefit of immune response modifiers (IRMs), which have shown promise in early clinical studies, is discussed. PMID- 11398807 TI - Homology of ab1 and ab3 monoclonal antibodies that neutralize Semliki Forest virus. AB - A noninternal image monoclonal antiidiotypic antibody (ab2 mAb), designated 1.13A321, that had proved its efficacy as vaccine against infection with Semliki Forest virus (SFV) in BALB/c mice, was used as immunogen to generate a panel of SFV-neutralizing monoclonal anti-anti-idiotypic antibodies (ab3 mAbs) to compare them genetically with ab1 mAb 1.13 (IgG2a). There are various studies that compare ab1 and ab3 mAbs but none that compare virus-neutralizing ab1 and ab3 mAbs. Five SFV-neutralizing ab3 MAbs, all IgG1, were obtained. The Vh gene (36 60), the D gene (Sp2), and the J gene (Jh2) encoding the heavy chain variable regions of all six mAbs, were similar and showed a high homology in the nucleotide sequence. The CDR3 amino acid sequences of four of five ab3 mAbs were identical to that of mAb1. One ab3 differed one amino acid in the CDR3 region. The results suggest that a strict selection criterion (virus neutralization) is sufficient to reach complete homology in the CDR3 region of mAb3. Future experiments are focused on selection of synthetic peptides in the CDR3 region as neutralizing mini-antibodies. PMID- 11398808 TI - Granulysin blocks replication of varicella-zoster virus and triggers apoptosis of infected cells. AB - Granulysin, a lytic protein present in cytolytic granules of human natural killer and cytotoxic T cells, entered cells infected with varicella-zoster virus (VZV). Exposure to granulysin accelerated death of infected cells as assessed by apoptosis markers. The functional domain of granulysin that mediated its antiviral effects was amino acid 23-51; this domain also mediates the additional antitumor cell effects of granulysin. Because granulysin is a product of natural killer cells and T lymphocytes, it is possible that its antiviral activity may act as a mediator of innate and adaptive immune mechanisms. PMID- 11398809 TI - Immunogenicity of two peptide determinants in the cytolytic T-cell response to flavivirus infection: inverse correlation between peptide affinity for MHC class I and T-cell precursor frequency. AB - We used the CD8+ cytotoxic T (Tc) cell immune response against the flavivirus, Murray Valley encephalitis virus (MVE), restricted by the H-2Kk major histocompatibility complex (MHC) class I molecule, to investigate immunodominance. Split-clone limiting dilution analysis revealed almost exclusive recognition of two peptides, MVE1785 and MVE1971, derived from the viral NS3 protein. The precursor frequency of MVE-reactive Tc cells was determined by limiting dilution analysis for cytotoxic function and intracellular staining for interferon-gamma; the latter gave a 100-fold higher estimate of MVE-reactive Tc cell precursors. MHC class I cell surface stabilization assays revealed that affinity for H-2Kk as well as halflives of the peptide-H-2Kk-complexes were markedly different for the two peptides. However, a kinetic study of antigen presentation showed that both peptides are presented for recognition by Tc cells with a comparable kinetics during the latent period of virus infection. Nevertheless, the lower affinity peptide MVE1785 elicited roughly twofold more Tc cell clones than the high-affinity peptide MVE1971. While the cytolytic activity against both determinants was similar after in vitro restimulation at the peak of the primary response, the smaller pool of memory anti-MVE1971 Tc cells correlated with an impaired memory response against that determinant, suggesting that the available T-cell repertoire is a major factor influencing the establishment of T cell memory. PMID- 11398810 TI - Identification of an immunodominant peptide in the parvovirus B19 VP1 unique region able to elicit a long-lasting immune response in humans. AB - The immune response against parvovirus B19 is mainly directed against the two structural proteins, VP1 and VP2. The amino terminal half of the VP1 unique region has been shown to elicit a dominant immune response in humans, more effective than other linear epitopes and also it has been seen to contain significant neutralizing linear epitopes. Three overlapping recombinant peptides corresponding to amino acids 2-40 (VP1-A), amino acids 32-71 (VP1-B), and amino acids 60-100 (VP1-C) of the VP1 unique region were produced by a procaryotic expression system. These peptides were used as antigens in a Western blot assay to detect specific immunoglobulin G (IgG) in serum samples from blood donors of different age groups with documented signs of a past B19 infection. Fragment VP1 C appeared significantly immunodominant over the other peptides, reacting with specific IgG in 86% of serum samples. The fragment VP1-C corresponds to a sequence with a known neutralizing activity and seems able to elicit a long lasting immune response because specific IgG were present in blood donors of all age groups. VP1-C would therefore appear to be an attractive candidate as a component of a subunit vaccine. PMID- 11398811 TI - Rapid and wide-reaching delivery of HIV-1 env DNA vaccine by intranasal administration. AB - Although the potential of DNA vaccination is now beginning to be greatly appreciated, no detailed study of its localization in tissue or its expression kinetics has been reported. In this study, we investigated these issues using HIV 1 DNA plasmids administered either intranasally or intramuscularly. Fluorescence in situ hybridization (FISH) revealed that the human immunodeficiency virus (HIV) plasmids administered intranasally localized in the alveoli, lung, liver, spleen, regional lymph nodes, kidney, fetus, and esophagus. These HIV plasmids were detected 2 to 4 weeks after administration. We detected messenger RNA production of HIV env gene in the lung, liver and spleen, and human immunodeficiency virus type 1 (HIV-1)-specific proteins were detectable in the lung. These observations may provide important information for understanding the mechanisms of strong immune activation induced by DNA vaccination via the intranasal route. This technology of DNA administration suggests possible practical applications for vaccination and probably for gene therapy. PMID- 11398812 TI - T-Cell immune activation in children with vertically transmitted hepatitis C virus infection. AB - Little is known concerning the clinical features, the histological outcome, and the effects on the maturation of immune system of children with vertically transmitted hepatitis C virus (HCV) infection. Specifically, no data are available on the peripheral distribution of T-cell subsets. The frequency of naive and memory cells, activated T cells, and cytokine-producing T cells was analyzed in nine HCV-infected children born to HCV-positive mothers. In HCV infected children, the distribution of naive and memory cells was not significantly altered in the CD4 subset whereas within the CD8 subset, an increase of memory and a decrease of naive cells was observed. The frequency of HLA-DR-positive and Fas-positive T cells was increased in HCV-infected children in both CD4 and CD8 subsets. The distribution of Fas-expressing T cells was directly related to that of HLA-DR cells and inversely related to the frequency of naive T cells. In regard with cytokine production we found increased levels of both CD4 and CD8 interferon-gamma (IFN-gamma)-producing cells whereas no difference in the percentage of interleukin-2 (IL-2)-producing T cells was observed. No meaningful correlation was observed between individual T cell subsets and ALT levels or HCV viral load. In conclusion, our results indicate an increased T-cell activation and a shift to a T(H)1 pattern of cytokine production in children with vertically transmitted HCV infection. The cause of this kind of immune response could reside in the persistent antigenic stimulation by chronic HCV infection. PMID- 11398813 TI - The role of interleukin-18 in vesicular stomatitis virus infection of the CNS. AB - Intranasal application of vesicular stomatitis virus (VSV) results in the initial infection of the olfactory receptor neurons and a rapid progression of the virus through the mouse central nervous system (CNS). Interleukin-18 (IL-18) is an 18.3 kd cytokine that induces interferon gamma (IFN-gamma) production in mice. IL-18 is synthesized as an inactive precursor that is cleaved and activated by caspase 1/interleukin-1beta converting enzyme (ICE). IL-18 shares several biological properties with IL-12, including the ability to induce IFN-gamma production in T lymphocytes and natural killer (NK) cells. In the CNS, microglia and astrocytes produce IL-18 and IL-12. We have previously shown that IL-12 promotes recovery from VSV encephalitis. This led us to examine the potential role of IL-18 in the pathogenesis of VSV encephalitis. We show that both IL-18 and caspase-1 mRNA are consistently present in the CNS of mice. The addition of exogenous IL-18 to cell cultures does not affect the production of VSV, and addition of exogenous IL-18 at the time of infection does not alter the morbidity or mortality of BALB/c mice. In vitro studies with neutralizing monoclonal antibody to IL-18 had no effect. From these results we conclude that in this system and under the experimental conditions used, unlike IL-12 and IFN-gamma, IL-18 does not play a significant role in the host response to VSV infection. PMID- 11398814 TI - Keystone symposium on Molecular Aspects of Viral Immunity: Keystone, Colorado, April 14-20, 2001. PMID- 11398815 TI - Tsl and LP-BM5: a comparison of two murine retrovirus models for HIV. AB - The ts1 murine leukemia virus produces an immunodeficiency state in mice that parallels human immunodeficiency virus (HIV) infection in humans. Other murine leukemia viruses, such as LP-BM5 used in the murine acquired immune deficiency virus (MAIDS) model, have been studied extensively as a small animal model for HIV research, but lack many key similarities to HIV. Mice infected with ts1, however, utilize CD4 target cells for infection, undergo neuronal loss and demyelination, and develop clinical immunodeficiency. These features make this retrovirus in many ways an ideal candidate for a small animal model for HIV research. In this review article, the early development, the molecular and clinical pathogenesis of both the ts1 mutant of the Moloney murine leukemia virus and LP-BM5 are examined. Based on an extensive evaluation of the literature on LP BM5 and ts1, it is concluded that the ts1 virus may serve as a better animal model to human retrovirus infection. PMID- 11398816 TI - Assessing the costs of school-based mental health services. PMID- 11398817 TI - Pseudoexfoliation and posterior capsular opacification. PMID- 11398818 TI - Retraction. PMID- 11398819 TI - Noise-induced hearing loss among U.S. Air Force cryptolinguists. AB - BACKGROUND: Occupational noise-induced hearing loss remains epidemic in the U.S. Air Force (USAF) and aircrew members continue to be affected. This study examines noise-induced hearing loss (NIHL) observed among USAF cryptolinguists that is attributable to radio noise and attempts to determine whether the current USAF Hearing Conservation Program (HCP) adequately identifies NIHL. METHODS: Audiograms from 120 ground-based cryptolinguists were examined. Comparisons were made between 1998 audiograms and either the reference audiogram or the enlistment audiogram. To determine HCP effectiveness, results were compared with the USAF standard of 3% or fewer permanent threshold shifts (PTS) per year. RESULTS: Some 13.3% of the cryptolinguists experienced standard threshold shifts (STS) between their enlistment audiogram and their initial occupational (reference) audiogram; 9.2% experienced STS in 1998 as compared with their initial enlistment audiogram but only 6 of the 11 (54%) were detectable using the reference audiogram as the baseline. The frequency pattern of changes in hearing thresholds was characteristic for NIHL. CONCLUSIONS: The NIHL that occurs among cryptolinguists prior to performance of the reference audiogram, and the hearing threshold shifts that occur between the enlistment audiogram and the reference audiogram, may obscure future hearing loss in the population. The incidence of PTS appeared to exceed 3% when the enlistment audiogram was used. While this result was not significantly different from 3%, sample size limitations and data accuracy concerns warrant that this population be closely watched in the future. PMID- 11398820 TI - Cases from the Aerospace Medicine Residents' Teaching File: acute onset of dizziness in a USAF aviator. AB - An Air Force F-15 pilot develops the sudden onset of incapacitating dizziness with nausea and vomiting not associated with flight. Presentation, diagnosis, treatment, and aeromedical issues surrounding the symptom complex of acute onset vertigo is discussed. PMID- 11398821 TI - A 39-year-old woman with an isolated pancreatic duct stricture. PMID- 11398825 TI - The growing family of NIH institutes. PMID- 11398826 TI - The growing family of NIH institutes. PMID- 11398827 TI - The growing family of NIH institutes. PMID- 11398828 TI - The Yanomamo and the 1960s measles epidemic. PMID- 11398830 TI - The Yanomamo and the 1960s measles epidemic. PMID- 11398831 TI - The Yanomamo and the 1960s measles epidemic. PMID- 11398832 TI - A beta-lactamase belonging to group 2e from oral clinical isolates of Prevotella intermedia. AB - A beta-lactamase in oral clinical isolates of Prevotella intermedia that hydrolyzed cefuroxime and cephalothin with rates of 600 and 53.3 respectively, relative to that for cephaloridine (100), was characterized as a 2e cephalosporinase. Inhibition was observed by clavulanic acid (IC50 0.72 microM), tazobactam (IC50 0.21 microM) and sulbactam (IC50 0.07 microM) and was not inhibited by cloxacillin, EDTA, NaCl or p-CMB. The pI and pH optima were 4.7 and 5.4, respectively. PMID- 11398833 TI - Alexander disease: new insights from genetics. AB - Prior to finding that GFAP mutations underlie many cases of Alexander disease, it was unclear whether the disease originated in astrocytes or if the formation of Rosenthal fibers was a response to an external insult. It was also unclear whether the etiology of the disease was environmental or genetic. For many cases of Alexander disease, these questions have now been answered. An immediate clinical benefit of this discovery is the possibility of diagnosing most cases of Alexander disease through analysis of patient DNA samples, rather than resorting to brain biopsy. In addition, fetal testing is now an option for parents who have had an Alexander disease child with an identified mutation and who wish to have additional children. For the future, these mutations should provide a unique window for illuminating the mechanism of the disease. PMID- 11398834 TI - Neuropathology of NFHgp160 transgenic mice expressing HIV-1 env protein in neurons. AB - The physiopathology of HIV-1 dementia remains largely hypothetical. Although several sets of evidence point towards an indirect multicellular inflammatory pathway, gp120, one of the HIV-1 env products, was shown to be very cytotoxic for neurons in vitro. To explore a direct pathway in the physiopathology of dementia in AIDS, we developed transgenic mouse models carrying the HIV-1 env proteins gp 120 and gp 41 (gp 160) under the control of the human light neurofilament and murine heavy neurofilament promoters. To date, this is the first mouse model in which the HIV-1 env protein can be detected in neurons by immunohistochemistry. The expression is found in several brainstem and spinal cord gray structures and in the cerebellum in one of the mouse lines bearing the NFHgp160 transgene. The morphological findings at 3 months are subtle and are dominated by a watery, dendritic degeneration and a reactive gliosis. At 12 months, the evidence of neuronal degeneration and loss is present along with various degenerative phenomena involving synapses, dendrites and axons, including axonal swellings. Cytoskeletal abnormalities were found by immunohistochemistry. Chronic inflammation was also observed in the leptomeninges of the spinal cord and brainstem and in the cerebellar white matter. These models thus offer an exciting sequence of morphological findings initiated by the neuronal expression of the HIV-1 env proteins and offer a different tool to explore the neuronal dysfunction in AIDS. PMID- 11398835 TI - The expression of intermediate filament protein nestin as related to vimentin and desmin in regenerating skeletal muscle. AB - Intermediate filament (IF) proteins show specific spatial and temporal expression during development of skeletal muscle. Nestin, the least known muscle IF, has an important role in neuronal regeneration. Therefore, we analyzed the expression pattern of nestin as related to that of vimentin and desmin during skeletal muscle regeneration. Nestin and vimentin appear at 6 h post-injury in myoblasts, with maximum expression around day 3-5 post-injury. Thereafter, vimentin expression ceases completely, whereas that of nestin is downregulated to remain only in the sarcoplasm next to neuromuscular and myotendinous junctions. Desmin appears at 6-12 h post-injury and becomes the predominant IF in myofibers simultaneously with the appearance of cross-striations. The expression pattern and colocalization of nestin and vimentin, known to form heteropolymers, suggests that they are essential during the early dynamic phase of the myofiber regeneration when migration, fusion, and structural modeling of myogenic cells occurs, whereas desmin is responsible for keeping myofibrils in register in mature myofibers. In conclusion, the expression of nestin is dynamically orchestrated with that of vimentin and desmin during skeletal muscle regeneration and recapitulates that seen during myogenesis, i.e. these IFs have key functional roles in the construction and restoration of skeletal myofibers. PMID- 11398836 TI - Distinct expression patterns and levels of enzymatic activity of matrix metalloproteinases and their inhibitors in primary brain tumors. AB - Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) have been implicated in the immense invasive potential and neovascularization of primary brain tumors. We investigated the gene expression profiles of MMPs 1, 2, 3, 7, 9, 12, 13, 14, 16 and of TIMPs 1, 2, 3, and 4 in various primary brain tumors (astrocytoma WHO grade I-III, glioblastoma, PNET, ependymoma III and oligoastrocytoma II) using novel RNase protection assay probe sets. In addition, we determined the level and cellular source of gelatinolytic activity and localized gelatinase B and TIMP-1 RNA. Distinct expression patterns of the MMP and TIMP genes were found in the various brain tumors tested. While the WHO grade I and II tumors had MT1/MT3 ratios below 1, the malignant (grade III and IV) tumors had ratios above 1. Strong expression of TIMP-1 RNA was observed in all malignant tumors and in grade I pilocytic astrocytomas and localized to the walls of neovessels. Quantitative analysis of enzymatic activity in the soluble fraction of protein extracts revealed that in most tumors gelatinases remained in the inactive pro-form. In situ zymography revealed net gelatinolytic activity in neurons of normal brain and in tumor cells and vessel walls of all tumors tested. Immunohistochemistry demonstrated that gelatinase B was localized to vessel walls, to neutrophils in areas of hemorrhage, and in glioblastomas to macrophages. Together these data demonstrate that the different primary brain tumors show distinct regulation of MMP and TIMP genes. The localization of the soluble gelatinase B indicates an association with neovascularization, whereas membrane-bound MMPs may account for the invasive potential of the glial tumor cells. PMID- 11398837 TI - Spiroplasma sp. 16S rDNA in Creutzfeldt-Jakob disease and scrapie as shown by PCR and DNA sequence analysis. AB - The pathogenesis of the transmissible spongiform encephalopathies (TSE), which include Creutzfeldt-Jakob disease (CJD) in humans and scrapie in sheep, remains an enigma. In this paper we present evidence for the association of Spiroplasma sp., a wall-less prokaryote, with TSE. We have shown PCR amplification of Spiroplasma 16S rDNA in TSE-infected brain tissues (13 of 13 CJD cases and 5 of 9 scrapie cases) and not in control brains (0 of 50). Direct sequencing of the amplified PCR products has confirmed the presence of Spiroplasma-like DNA in all 5 of the TSE brains tested. Our evidence is not necessarily in conflict with involvement of a PrPres--a protease-resistant host-derived protein referred to as the prion--in the pathogenesis of TSE, since there is evidence that another factor is involved. We propose a bacterium, namely Spiroplasma, as this associated factor although the role of Spiroplasma in TSE cannot be determined from these experiments. The presence of the nucleic acid sequence of this microbe in all cases of TSE in our laboratory and not in controls provides direct evidence of the association of Spiroplasma sp. with TSE. PMID- 11398838 TI - Mouse model of Bell's palsy induced by reactivation of herpes simplex virus type 1. AB - In order to investigate the mechanism of Bell's palsy, we developed an animal model of facial nerve paralysis induced by the reactivation of herpes simplex virus type 1 (HSV-1). Eight weeks after recovery from facial nerve paralysis caused by inoculation with HSV-1, the mice were treated with auricular skin scratch at the site of the previous inoculation, or with intraperitoneal injection of anti-CD3 monoclonal antibody (mAb), or combination of both procedures. No mice developed facial nerve paralysis when they were treated with either auricular scratch or mAb injection alone. In contrast, 20% of mice developed facial nerve paralysis with the combined treatment. With one exception, no mouse treated with either auricular scratch or mAb injection showed HSV-I DNA in their facial nerve tissue, whereas 4 out of 6 mice receiving both treatments showed HSV-1 DNA on day 10 after treatment. Histopathological findings showed neuronal degeneration in the geniculate ganglion and demyelination of the facial motor nerve in paralyzed mice. These findings suggest that a combination of stimuli, local skin irritation, and general immunosuppression is essential for successfully inducing facial nerve paralysis in mice with latent HSV-1 infection. PMID- 11398839 TI - Chromosome 1p and 14q FISH analysis in clinicopathologic subsets of meningioma: diagnostic and prognostic implications. AB - The second most frequently reported genetic abnormalities in meningiomas after 22q loss are deletions of 1p and 14q. To assess the potential diagnostic and prognostic utility of these chromosomal alterations, we studied 180 well characterized meningiomas using dual-color fluorescence in situ hybridization (FISH) with DNA probes localized to 1p32, 1p36, 14q13, and 14q32. Our cohort consisted of 77 benign (grade I), 74 atypical (grade II), and 29 anaplastic (grade III) meningiomas. Benign and atypical meningiomas were further stratified into subsets of recurring (despite gross total resection) vs non-recurring (at least 10 yr of follow-up) and mitotically active vs brain invasive subsets, respectively. Losses of 1p and 14q losses were identified in 23% and 31% of benign, 56% and 57% of atypical, and 75% and 67% of anaplastic meningiomas, respectively (p < 0.001 for 1p; p = 0.004 for 14q). Combined 1p/14q deletions were encountered in 7% benign. 39% atypical, and 63% anaplastic meningiomas (p < 0.001). Benign non-recurring meningiomas were less likely to harbor 14q deletions than recurring examples (17% vs 50%, p = 0.013). There was a trend for anaplastic meningiomas with 14q deletions and atypical meningiomas with combined 1p/14q deletions to have poorer overall survivals, though neither reached statistical significance. We conclude that 1p and 14q deletions are highly associated with increasing histologic grade and play an important role in meningioma tumor progression. Furthermore, 14q FISH analysis may aid in assessing recurrence risk in histologically benign meningiomas. PMID- 11398840 TI - Dominance of autoreactive T cell-mediated delayed-type hypersensitivity or antibody-mediated demyelination results in distinct forms of experimental autoimmune neuritis in the Lewis rat. AB - The role of anti-myelin antibodies in the pathogenesis of experimental autoimmune neuritis (EAN) induced in the Lewis rat by immunization with peripheral nerve myelin has been assessed. Passive transfer with lymph node cells (LNC) or purified serum immunoglobulin from rats with EAN was employed to directly measure the contribution of B cells and anti-myelin antibodies to demyelination and disease. Lewis rats with EAN transferred by LNC or purified serum immunoglobulin from EAN donors in conjunction with a low dose of P2-specific CD4+ T cells demonstrated profound histopathological and neurophysiological evidence of demyelination during disease. In contrast, the classical adoptive transfer model of EAN in the Lewis rat induced by the injection of P2-specific CD4+ T cells was characterized by histopathological and neurophysiological evidence of axonal dysfunction and degeneration with limited demyelination. These findings demonstrate that the synergistic action of T cells and anti-myelin antibodies mediating demyelination or purely T cell mediated axonal dysfunction and degeneration are distinct pathways by which a specific autoimmune response in the peripheral nervous system can cause neurological disease. PMID- 11398841 TI - Microglial activation parallels system degeneration in progressive supranuclear palsy and corticobasal degeneration. AB - The role of microglia in progressive supranuclear palsy (PSP) and corticobasal degeneration (CBD) is unknown. To address this issue we examined 10 cases of PSP, 5 cases of CBD, and 4 normal controls. Microglial and tau burdens were determined with image analysis on brain sections that had been immunostained with monoclonal antibodies to HLA-DR and phospho-tau. We found that microglial activation was greater in PSP and CBD than normal controls, and that the microglial burden correlated with the tau burden in most areas. There were distinct patterns of microglial activation and tau pathology in PSP and CBD, with PSP showing more pathology in infratentorial structures and CBD showing more pathology in supratentorial structures. These results support the notion that PSP and CBD are distinct clinicopathologic entities. Microglial activation was not well correlated with tau pathology in the brainstem of PSP, which suggests that brainstem pathology in PSP is not exclusively due to tau pathology. While the results do not necessarily support a direct causal link between microglial activation and neurodegeneration in PSP or CBD, they nevertheless suggest that microglia play a role in disease pathogenesis. PMID- 11398842 TI - Neuropathologic substrates of ischemic vascular dementia. PMID- 11398843 TI - Detection of c-fos expression in benign and malignant musculoskeletal lesions. AB - The proto-oncogene c-fos has been implicated in the development of both benign and malignant lesions of bone. Although c-fos expression in such lesions has been well studied in transgenic mouse models, less is known about its role in human musculoskeletal pathology. To clarify this relationship, we used in situ hybridization to localize c-fos m-RNA transcripts in 26 fibrous lesions (eight cases of extra-abdominal fibromatosis and six cases each of fibrous dysplasia, fibrosarcoma, and malignant fibrous histiocytoma of bone) as well as six chondrosarcomas and eight conventional high grade osteosarcomas. We found detectable levels of c-fos expression in tissues from each type of lesion tested. Moreover, all fibrous lesions consistently demonstrated high levels of expression in a majority of cells in each lesion. Chondrosarcomas and osteosarcomas exhibited more heterogeneity in c-fos expression than fibrous tissues. Three of six chondrosarcomas showed moderate expression of c-fos while only one of six was considered high. Similarly, only three of eight osteosarcomas had high expression of c-fos. These findings indicate that the expression of c-fos may be important in the development of a broad range of fibrous lesions as well as in bone and cartilaginous tumors. Additionally, this is the first report, to our knowledge, of detectable c-fos m-RNA in human chondrosarcoma. PMID- 11398844 TI - Expression of a newly defined tumor-rejection antigen SART3 in musculoskeletal tumors and induction of HLA class I-restricted cytotoxic T lymphocytes by SART3 derived peptides. AB - We recently reported that a SART3 tumor-rejection antigen possessing tumor epitopes is capable of inducing HLA class 1-restricted and tumor-specific cytotoxic T lymphocytes in cancer patients. We studied the expression of the SART3 protein in musculoskeletal tumors to find a molecule for potential use in tumor-specific immunotherapy. The SART3 was detected at protein levels in 100% of the osteosarcoma cell lines (n = 20), in 50% of the musculoskeletal tumor tissue specimens (n = 32), and at notable levels in 67% of osteosarcoma tissues (n = 9) and malignant fibrous histiocytosis tissues (n = 9), respectively. SART3-derived peptides at positions 109-118 and 315-323 induced HLA-A24-restricted tumor specific cytoxic T lymphocytes from peripheral blood mononuclear cells of patients with osteosarcoma or malignant fibrous histiocytosis. These peptide induced cytotoxic T lymphocytes recognized HLA-A24+ SART3+ osteosarcoma cells but not HLA-A24 or SART3 cells. These results suggest that the SART3 protein and its derived peptides could be molecules appropriate for use in specific immunotherapies for approximately 60% of HLA-A24+ patients with osteosarcoma or malignant fibrous histiocytosis. PMID- 11398845 TI - Growth in width of the metacarpals--an investigation in human fetuses. AB - The process of growth in width of the human hand during fetal life has never been described. Do metacarpals grow concentrically and separation between the bones occurs through expansion of soft tissues? Or is growth eccentric, a process termed drift by Enlow, a relocation in space of organs? Hands of 10 spontaneously aborted fetuses (age range: between 14.5 and 24 weeks of gestation) were examined paying special attention to the bone bark. A thicker bone bark was taken as an indication of growth in that direction. The thickness of the bone bark was measured at the radial and ulnar sides at the level of the proximal and of the distal physes of the second to fifth metacarpals. A ratio of radial over ulnar bone bark thickness (R/U ratio) was calculated. The third metacarpal grew almost concentrically (R/U ratio 1.12 +/- 0.06). The second metacarpal grew in a radial direction (R/U ratio 3.29 +/- 0.19) and the fourth and more so the fifth metacarpal grew in an ulnar direction (R/U ratio 0.70 +/- 0.04 and 0.42 +/- 0.02, respectively). The differences in R/U ratios between every metacarpal were statistically significant for all comparisons P < or = 0.001. Fetal growth in width of the human metacarpals is eccentric and not concentric. It is concluded that during growth in width the metacarpals move away from the midline of the hand and that growth occurs through eccentric bone apposition rather than through soft tissue expansion. PMID- 11398846 TI - Regional material properties of the human hip joint capsule ligaments. AB - The hip joint capsule functions to constrain translation between the femur and acetabulum while allowing rotational and planar movements. Despite the crucial role it plays in the pathogenesis of hip instability, little is known about its biomechanical properties. The goal of this study was to determine the regional material properties of the iliofemoral and ischiofemoral ligaments of the capsule. Ten human cadaveric specimens of each ligament were tested to failure in tension. The stress at failure, strain at failure, strain energy density at failure, toe- and linear-region elastic moduli, and the Poisson's ratio were measured for each ligament. The strain to failure was greatest in the ischiofemoral ligament, while no significant difference was noted in failure stress by region or ligament. The Young's moduli of elasticity ranged from 76.1 to 285.8 MPa among the different ligaments, and were generally consistent with properties previously reported for the shoulder capsule. The elastic moduli and strain energy density at failure differed by region. No significant differences in Poisson's ratio were found by region or ligament. The average Poisson's ratio was approximately 1.4, consistent with anisotropic behavior of ligamentous tissues. Understanding the material properties of the hip capsule may help the orthopaedic surgeon better understand normal ligament function, and thereby choose a surgical approach or strategy of repair. Furthermore, knowledge of the normal mechanical function of the hip capsule ligaments could assist in the evaluation of the success of a repair. PMID- 11398847 TI - GDF-5 deficiency in mice alters the ultrastructure, mechanical properties and composition of the Achilles tendon. AB - Acromesomelic dysplasia of the Hunter-Thompson and Grebe types are rare human disorders based on growth/differentiation factor (GDF)-5/CDMP-1 genetic mutations. Numerous skeletal abnormalities are present in these individuals, including shortened limb bones and severe dislocations of the knee. In the GDF-5 deficient brachypodism mouse, similar, although less severe, phenotypes are observed. It is unknown whether the joint dislocations observed in these disorders are due to a defect in the original formation of joints such as the knee, or to abnormalities in the tendons and ligaments themselves. We hypothesized that tendons from GDF-5 deficient mice would exhibit altered composition, mechanical properties, and ultrastructure when compared with heterozygous control littermates. GDF-5 deficient Achilles tendons were structurally weaker than controls, and structural strength differences appeared to be caused by compromised material properties: after normalizing by collagen per unit length, mutant tendons were still 50% weaker (P < 0.0001) and 50% more compliant (P < 0.001) than controls. Despite comparable levels of skeletal maturity in the two cohorts, the majority of mutant tendon failures occurred in the mid-substance of the tendon (64% of all failures), whereas the majority of control failures occurred via avulsion (92% of all failures). Mutant Achilles tendons contained 40% less collagen per microgram of DNA when compared to controls (P = 0.004). No significant difference in glycosaminoglycan (GAG)/DNA was detected. Ultrastructural analyses indicated a slight trend toward increased frequency of small diameter (30-100 nm) collagen fibrils in the mutant Achilles. Our findings suggest that increased tendon and ligament laxity may be the cause of the joint dislocations seen in patients with Hunter-Thompson and Grebe type dysplasia, rather than developmental abnormalities in the joints themselves. PMID- 11398848 TI - Autonomic innervation of tendons, ligaments and joint capsules. A morphologic and quantitative study in the rat. AB - We analyzed the neuronal occurrence of autonomic transmitters; noradrenaline (NA), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP), in the Achilles tendon, medial and lateral collateral ligaments and knee joint capsule in the rat--by immunohistochemistry (IHC). In addition, the tissue concentrations of the sympathetic neuropeptide, NPY, and the parasympathetic peptide, VIP, were determined by radioimmunoassay (RIA). IHC demonstrated nerve fibers containing sympathetic vasoconstrictors--NA and NPY--and the parasympathetic vasodilator, VIP, in all tissues. NPY- and NA-positive nerve fibers were predominantly observed in larger blood vessels, whereas, nerve fibers immunoreactive to VIP were found in smaller vessels. In many nerve fibers a co-localization of the transmitters was seen. RIA showed that the concentration of NPY compared to VIP was 15-times higher in ligaments and twice as high in tendons and capsules. The differences noted may reflect a difference in vulnerability to degenerative conditions. In pathological conditions, dysregulation of autonomic transmitters in hypovascularized tissues subjected to repetitive mechanical load may contribute to tissue hypoxia leading to degeneration and rupture of tendons and ligaments. PMID- 11398849 TI - Origin and pathway of sensory nerve fibers to the ventral and dorsal sides of the sacroiliac joint in rats. AB - The purpose of this study is to clarify sensory innervation in the ventral and dorsal sides of the sacroiliac joint. Fluoro-gold, a neural tracer, was injected into the left sacroiliac joint of adult rats from the dorsal side after denervation of the dorsal side, and the bilateral dorsal root ganglia (DRGs) from T13 to S4 were examined by fluorescence microscopy five days after injection. In another rat group, the DRGs were examined using the same methods after injection of fluoro-gold from the ventral side. In the case of dorsal denervation (ventral nerve supply), labeled neurons were mainly located in the ipsilateral DRGs from L1 to S2. On the other hand, in the case of ventral denervation (dorsal nerve supply), labeled neurons were noted in the ipsilateral DRGs from L4 to S2. The sacroiliac joint in rats is innervated differently on the ventral and dorsal sides: the sensory nerve fibers to the dorsal side of the sacroiliac joint were derived from the DRGs of lower lumbar and sacral levels (from L4 to S2); and those to the ventral side from the DRGs of upper lumbar, lower lumbar, and sacral levels (from L1 to S2). PMID- 11398850 TI - Vascular response of the meniscus to injury: effects of immobilization. AB - Failed meniscal healing may lead to degenerative osteoarthritis of the knee. Healing is thought to be dependent upon an adequate blood supply, yet "normal" vascular changes during healing are not well understood. In this study we have quantified vasoactive and angiogenic responses to medial meniscal injury in a rabbit model under clinically relevant conditions, and related these to histological criteria of healing. Twenty-six adult rabbits were given a standardized meniscal injury; 12 of these had the hind limb immobilized by pinning. Eight normal controls and 12 sham-operated animals were also studied. After 4 weeks, animals underwent either vascular volume (vascular index) determination, or blood flow measurement using coloured microspheres. Histological analysis was also performed to assess meniscal healing. In injured animals, blood flow to the menisci was increased fivefold 4 weeks post-operative; this increase was prevented by immobilization. The vascular index of the menisci was also increased threefold by injury, but not significantly reduced by immobilization. Histological examination of injured menisci showed examples of healing and non-healing tears in both mobile and immobile groups. Meniscal injuries are associated with characteristic changes in vascularity and perfusion, and these changes likely play a significant role in the healing process. Characterization of the vascular responses to meniscal injury may lead to techniques that can promote reliable healing of meniscal tears and thereby improve clinical outcomes. PMID- 11398851 TI - Effects of oophorectomy on functional properties of resistance arteries isolated from the cancellous bone of the rabbit femur. AB - Information is sparse concerning the effect of oophorectomy (OOX) on bone vascularization and blood flow of possible significance for altered remodeling. Whether OOX affects functional characteristics of isolated bone resistance arteries was investigated. Ring preparations (diameter approximately 250 microm) of small femoral bone arteries from oophorectomized and sham-operated rabbits were mounted on a myograph six weeks postoperatively. Cumulative concentration response curves were obtained for various agonists at a normalized lumen diameter. Oophorectomy did not significantly influence lumen diameter, maximal response to high potassium, or maximal response to high potassium and 10(-5) M noradrenaline. However, OOX significantly increased the maximal response to noradrenaline (OOX 2.14 +/- 0.36 N/m, Sham 1.25 +/- 0.14 N/m) and endothelin-1 (OOX 1.76 +/- 0.32, Sham 0.95 +/- 0.15) in metaphyseal arteries. Moreover, the corresponding maximal active pressure for the agonists was significantly increased. OOX did not influence endothelial function assessed by the effects of acetylcholine or substance P. The functional responses of diaphyseal arteries were unaffected by OOX. The study demonstrates regional differences in the effects of OOX on small arteries of importance for control of vascular resistance in bone which suggests a relation between altered vascular function after ovarian hormonal withdrawal and the changes in bone turnover associated with osteoporosis. PMID- 11398852 TI - DL-penicillamine induced alteration of elastic fibers of periosteum-perichondrium and associated growth inhibition: an experimental study. AB - Perichondrium-periosteum, being of collagen and elastic fiber, is regarded as a bone growth regulating factor. The aim of the present study was to investigate the role of collagen and elastic fibers on bone growth, by interfering with the fiber assembly in growing chicks upon administration of DL-penicillamine (DL PNA). Our findings demonstrated that DL-PNA determined relevant modifications in the perichondrium-periosteum, as shown by histochemical, histomorphometrical,biochemical and ultrastructural analysis. This chemical has been shown to inhibit the formation of desmosine cross-links in elastin and to induce an increase of elastin associated microfibrils. On the contrary, the collagen network and the biochemical collagen markers were not affected. These changes resulted in a dramatically reduced growth of long bones in comparison with control. Perichondrial-periosteal regulation of bone growth may be mediated by mechanical and biological factors. This study demonstrates a microstructural change in the perichondrium-periosteum with decreased elastin and increased elastic microfibrils content in penicillamine treated chicks. The mechanism linking changes in the perichondrium-periosteum with altered growth still needs to be elucidated. PMID- 11398853 TI - Histomorphological and proliferative characterization of developing periosteal neochondrocytes in vitro. AB - Periosteal chondrogenesis is relevant to cartilage repair and fracture healing. Cell proliferation is a limiting factor of cartilage production. We used an in vitro organ culture model to test the hypothesis that proliferative activity correlates with cell morphology. One hundred and four periosteal explants from 26 two-month old New Zealand rabbits were cultured for up to 42 days. They were analyzed histomorphologically, and immunohistochemically with proliferative cell nuclear antigen (PCNA). The periosteal neocartilage displayed a consistent zonal pattern of chondrocyte cell shapes. The flat cell zone from day 7 to 21, consisted of uniform-sized small spindle-shaped cells. The round cell zone, which appeared on day 14, consisted of variable-sized round cells averaging 510 +/- 250 microm2 in area. They were subdivided into small round (<510 microm2) and large round cells (>510 microm2). The proliferative index was highest in the small round cell group (32 +/- 6%), intermediate in the flat cell group (27 +/- 6%), and lowest in the large round cell group (20 +/- 7%) (P < 0.001). Furthermore, the proliferative indices in the round cell group were inversely proportional to cell size. Therefore, (1) there is a sequential progression of cell morphology during periosteal chondrogenesis, (2) cell differentiation is arrested prior to terminal differentiation for some cells and not for others, and (3) proliferative activity is strongly related to cell morphology. This organ culture model provides us with opportunities to study the regulation of terminal chondrocyte differentiation and the control of cell proliferation. This will contribute to our understanding of cartilage repair, fracture healing and growth plate physiology. PMID- 11398854 TI - Fatigue of cortical bone under combined axial-torsional loading. AB - The influence of torsional loading on the fatigue life of cortical bone was investigated by conducting in vitro testing. Fatigue tests were conducted on cylindrical dumbbell bovine cortical bone specimens in an environmental chamber under axial loading, torsional loading and various combinations of axial torsional loading where the phase relationship and relative magnitudes of axial and torsional loadings were systematically varied. It was found that the superposition of torque on axial loading reduced the fatigue life of cortical bone. The reduction in fatigue life was significant when the maximum shear stress was greater than 59% of the maximum normal stress. The magnitude of reduction in the fatigue life of bone at low as well as high levels of axial loading depended on the magnitude of torsional loading, but was independent of the phase angle by which the torsional loading lagged the axial loading. Furthermore, oblique fracture profiles, characteristic of torsion-induced failure, were observed for combined axial-torsional load cases. Based on these results, it is suggested that torsional loading plays a significant role in determining the fatigue life of cortical bone. PMID- 11398856 TI - Age and ovariectomy impair both the normalization of mechanical properties and the accretion of mineral by the fracture callus in rats. AB - The impact of age and ovariectomy on the healing of femoral fractures was studied in three groups of female rats at 8, 32 and 50 weeks of age at fracture. In the two older groups, the rats had been subjected to ovariectomy or sham surgery at random at 26 weeks of age. At fracture, all rats received unilateral intramedullary pinning of one femur and a middiaphyseal fracture. Rigidity and breaking load of the femora were evaluated at varying times up to 24 weeks after fracture induction by three-point bending to failure. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry. In the youngest group, 8-week old female rats regained normal femoral rigidity and breaking load by 4 weeks after fracture. They exceeded normal contralateral values by 8 weeks after fracture. In the middle group, at 32 weeks of age, fractures were induced, and the femora were harvested at 6 and 12 weeks after fracture. At 6 weeks after fracture there was partial restoration of rigidity and breaking load. At 12 weeks after fracture, only the sham-operated rats had regained normal biomechanical values in their fractured femora, while the fractured femora of the ovariectomized rats remained significantly lower in both rigidity and breaking load. In contrast, for the oldest group of rats, 50 weeks old at fracture, neither sham-operated nor ovariectomized rats regained normal rigidity or breaking load in their fractured femora within the 24 weeks in which they were studied. In all fractured bones, there was a significant increase in BMD over the contralateral intact femora due to the increased bone tissue and bone mineral in the fracture callus. Ovariectomy significantly reduced the BMD of the intact femora and also reduced the gain in BMD by the fractured femora. In conclusion, age and ovariectomy significantly impair the process of fracture healing in female rats as judged by measurements of rigidity and breaking load in three point bending and by accretion of mineral into the fracture callus. PMID- 11398855 TI - The effect of aging on distraction osteogenesis in the rat. AB - The effect of age on bone formation in the limb lengthening model of distraction osteogenesis (DO) was investigated in two studies using Sprague-Dawley (SD) rats from two colonies at various ages (CAMM: 9 vs 24 months, Harlan: 4 vs 24 months). External fixators were placed on the right tibiae of 30 male SD rats (20 CAMM, 10 Harlan) and mid-diaphyseal osteotomies were performed. Distraction was performed at 0.2 mm bid for 20 days (CAMM) or 14 days (Harlan). The experimental (DO) and control (contra-lateral) tibiae were removed for high-resolution radiography and decalcified histology. Videomicroscopy was used to quantitate radiodensity, histology (matrix type) and relative areas of cell proliferation, which was identified by proliferating cell nuclear antigen (PCNA) immunochemistry. Both studies demonstrated an age-related decrease in the percent mineralized bone (radiodensity) in the distraction gap (CAMM 9 vs 24 months: 68% vs 51%, P < 0.003; Harlan 4 vs 24 months: 95% vs 36%, P < 0.001) and no significant colony or distraction time-specific difference was seen between the two colonies of 24 month-old rats. Histology was performed on the Harlan rats. The DO gaps in the 24 month-old rats demonstrated less endosteal new bone compared to the 4-month-old rats (P < 0.01), but equivalent periosteal new bone. In 4-month-old rats, PCNA immunostained cells were organized along the primary matrix front (where the first deposition of osteoid occurs) extending across both periosteal and endosteal surfaces. In 24-month-old rats, PCNA+ cells were organized in zones along the periosteal new bone fronts only and irregularly scattered throughout the endosteal gap within a fibrovascular non-ossifying matrix. These results indicate that 24-month-old rats have a relative deficit in endosteal bone formation which may not be related to cell proliferation but rather to cell organization. This model reflects the clinical situation where radiographic findings in older patients demonstrate significant delays in mineralization during DO. We believe this model of DO in aged rats presents unique in vivo opportunities to test hypotheses concerning (1) the effects of aging on bone repair, (2) the effects of pharmacological agents on bone repair in a geriatric setting, and (3) to study the mechanisms underlying DO. PMID- 11398857 TI - Ischemia causes muscle fatigue. AB - The purpose of this investigation was to determine whether ischemia, which reduces oxygenation in the extensor carpi radialis (ECR) muscle, causes a reduction in muscle force production. In eight subjects, muscle oxygenation (TO2) of the right ECR was measured noninvasively and continuously using near infrared spectroscopy (NIRS) while muscle twitch force was elicited by transcutaneous electrical stimulation (1 Hz, 0.1 ms). Baseline measurements of blood volume, muscle oxygenation and twitch force were recorded continuously, then a tourniquet on the upper arm was inflated to one of five different pressure levels: 20, 40, 60 mm Hg (randomized order) and diastolic (69 +/- 9.8 mm Hg) and systolic (106 +/ 12.8 mm Hg) blood pressures. Each pressure level was maintained for 3-5 min, and was followed by a recovery period sufficient to allow measurements to return to baseline. For each respective tourniquet pressure level, mean TO2 decreased from resting baseline (100% TO2) to 99 +/- 1.2% (SEM), 96 +/- 1.9%, 93 +/- 2.8%, 90 +/ 2.5%, and 86 +/- 2.7%, and mean twitch force decreased from resting baseline (100% force) to 99 +/- 0.7% (SEM), 96 +/- 2.7%, 93 +/- 3.1%, 88 +/- 3.2%, and 86 +/- 2.6%. Muscle oxygenation and twitch force at 60 mm Hg tourniquet compression and above were significantly lower (P < 0.05) than baseline value. Reduced twitch force was correlated in a dose-dependent manner with reduced muscle oxygenation (r = 0.78, P < 0.001). Although the correlation does not prove causation, the results indicate that ischemia leading to a 7% or greater reduction in muscle oxygenation causes decreased muscle force production in the forearm extensor muscle. Thus, ischemia associated with a modest decline in TO2 causes muscle fatigue. PMID- 11398858 TI - Carrageenan-induced subacromial bursitis caused changes in the rat's rotator cuff. AB - This study was designed to investigate the histologic expression of the rat's supra- and infraspinatus tendons in carrageenan-induced subacromial bursitis. Thirty-two rats received subacromial injections with carrageenan (n = 28) or saline (n = 4). The tendons were analysed microscopically after staining with hematoxyline eosin, Van Giesons hematoxyline and immunofluorescent staining of fibronectin and fibrinogen. In the controls (saline x 10) and group A (carrageenan x 5) there were no changes in the tendons. In group B (carrageenan x 10) 3/8 rats showed macrophages between the collagen fibres and an increased staining of fibronectin. In group C (double dosis carrageenan) all rats had signs of fibrocartilaginous metaplasia in the supraspinatus tendon. In eight of these specimens even bony metaplasia was seen. The infraspinatus tendon showed fibrosis but no fibrocartilaginous metaplasia. The results showed that iatrogenic bursitis after carrageenan subacromial injections was associated with marked changes of the supraspinatus tendon. PMID- 11398859 TI - Vascular endothelial growth factor (VEGF) expression in the subacromial bursa is increased in patients with impingement syndrome. AB - Vascular endothelial growth factor (VEGF), which is known to be an angiogenetic factor, plays an important role in the inflammation of synovial tissue. To investigate the relationships between VEGF and clinical symptoms in rotator cuff disease, VEGF expression was examined using RT-PCR and immunohistochemical analysis in 50 patients with this disease (26 with full-thickness cuff tear, 12 with partial-thickness tear, and 12 with subacromial bursitis). VEGF mRNA expression was detected in 40 out of 50 patients by RT-PCR. VEGF mRNA expression was found more frequently in the patients with motion pain (39 out of 41) than in those without motion pain (1 out of 9) with statistical significance (Fisher's test, P < 0.001). Thirty-one out of 33 patients with synovial proliferation showed VEGF mRNA expression, whereas the expression of this transcript was found in 9 out of 17 patients without synovial proliferation. This association with synovial proliferation was also significant (Fisher's test, P = 0.0013). Thirty out of 41 patients with motion pain had synovial proliferation but 3 out of 9 patients without motion pain had synovial proliferation. In all these 30 patients with both motion pain and synovial proliferation, VEGF mRNA expression was detected. This association between motion pain and synovial proliferation was also significant (Fisher's test, P < 0.05). The mean vessel count and area in subacromial bursa expressing VEGF was significantly higher than in those without VEGF (Mann Whitney's U test, P < 0.01). These results suggested that VEGF expression is associated with vascularity, synovial proliferation and shoulder motion pain in the rotator cuff disease. PMID- 11398860 TI - Suppression of fibrous adhesion by proteoglycan decorin. AB - Small proteoglycan decorin is known to suppress the bioactivity of TGF-beta through a competitive binding with the cell surface receptors for the cytokine. Based on this knowledge, we hypothesized that decorin could reduce the formation of fibrous adhesion, because our previous study showed the neutralizing antibody to TGF-beta1 has that effect. An intra-articular adhesion model in the rabbit knee joint was employed in this study, and decorin was administered into the joint cavity continuously during the 4 weeks of the experiment. The results of the dose-response study demonstrated that decorin suppresses formation of fibrous adhesion in a dose-dependent manner. When the administration of decorin was limited to shorter periods, this effect was considerably impaired and the necessity of long-term administration was demonstrated. On the other hand, when administered together with TGF-beta1, decorin still suppressed adhesion but to a lesser extent, and it was suggested that this proteoglycan could have other significant mechanism(s) to suppress adhesion besides the neutralization of TGF beta. Thus, the present study showed that decorin could inhibit adhesion formation by both TGF-beta dependent and independent mechanisms. Considering that decorin exists ubiquitously in the body, its administration might be a promising approach to suppress adhesion. PMID- 11398861 TI - Functional roles of abdominal and back muscles during isometric axial rotation of the trunk. AB - Electromyographic (EMG) studies have shown that a large number of trunk muscles are recruited during axial rotation. The functional roles of these trunk muscles in axial rotation are multiple and have not been well investigated. In addition, there is no information on the coupling torque at different exertion levels during axial rotation. The aim of the study was to investigate the functional roles of rectus abdominis, external oblique, internal oblique, latissimus dorsi, iliocostalis lumborum and multifidus during isometric right and left axial rotation at 100%, 70%, 50% and 30% maximum voluntary contractions (MVC) in a standing position. The coupling torques in sagittal and coronal planes were measured during axial rotation to examine the coupling nature of torque at different levels of exertions. Results showed that the coupled sagittal torque switches from nil to flexion at maximum exertion of axial rotation. Generally, higher EMG activities were shown at higher exertion levels for all the trunk muscles. Significant differences in activity between the right and left axial rotation exertions were demonstrated in external oblique, internal oblique, latissimus dorsi and iliocostalis lumborum while no difference was shown in rectus abdominis and multifidus. These results demonstrated the different functional roles of trunk muscles during axial rotation. This is important considering that the abdominal and back muscles not only produce torque but also maintain the spinal posture and stability during axial rotation exertions. The changing coupling torque direction in the sagittal plane when submaximal to maximal exertions were compared may indicate the complex nature of the kinetic coupling of trunk muscles. PMID- 11398862 TI - Roles of thromboxane A2 and leukotriene B4 in radicular pain induced by herniated nucleus pulposus. AB - Biologically active substances, such as prostaglandins, thromboxanes, and leukotrienes, which are metabolites involved in the arachidonic acid cascade, are detected in herniated disc samples obtained from patients with lumbar disc herniation. However, little is known concerning the relationships between these substances and clinical symptoms such as radicular pain. Thromboxane A2 (TXA2) induces not only potent platelet aggregation, but also blood vessel contraction. Leukotriene B4 (LTB4), a potent chemotactic agent, plays a role in inflammatory reactions by recruiting neutrophils and lymphocytes. The purpose of this study was to examine the roles of TXA2 and LTB4 in the hyperalgesia induced by application of nucleus pulposus to the lumbar nerve root in the rat. TXA2 synthetase inhibitor and LTB4 receptor antagonist, which were injected into the epidural space, decreased mechanical hyperalgesia at both three and seven days after epidural injection. There were no significant differences in sensitivity to noxious thermal stimuli following application of the nucleus pulposus or an epidural injection. Epidural injection of LTB4 receptor antagonist and/or TXA2 synthetase inhibitor may attenuate the painful radiculopathy due to lumbar disc herniation. In conclusion, our findings suggest that TXA2 and LTB4 may play significant roles in mechanical hyperalgesia induced by autologous nucleus pulposus. PMID- 11398863 TI - Isolation of RNA from small human articular cartilage specimens allows quantification of mRNA expression levels in local articular cartilage defects. AB - Human adult cartilage is an inherently difficult tissue from which to isolate RNA. The RNA isolation techniques described so far have generally only been successfully applied to the isolation of RNA from larger amounts of cartilage. However, it is important to be able to analyse focal cartilage lesions in order to understand the local processes in the cartilage degeneration process. Therefore, we have developed a protocol for isolating RNA directly from as little as 10 mg wet weight of cartilage followed by quantitative PCR analysis. We were able to analyse the expression levels of several genes in parallel including aggrecan and type II collagen. PMID- 11398864 TI - Autologous chondrocyte implantation in a canine model: change in composition of reparative tissue with time. AB - The objective of the study was to evaluate the tissue types filling 4-mm diameter defects in the canine trochlear groove 1.5, 3, and 6 months after autologous chondrocyte implantation (ACI). Untreated defects served as controls. Periosteum alone controls were also included at the 1.5-month time period. The results were compared with previously published findings obtained 12 and 18 months postoperative. After 3 months the ACI-treated defects contained significantly more reparative tissue than found in the untreated control group, including twice the amount of hyaline cartilage (HC). These findings, however, were the only significant effects of the ACI treatment when compared to the periosteum alone or empty control groups. The benefits of ACI found at 3 months did not persist to longer time periods. An evaluation of the inter-observer error associated with the histomorphometric method indicated that it was generally less than the inter animal variation in the results. PMID- 11398865 TI - Development of mechanically stable alginate/chondrocyte constructs: effects of guluronic acid content and matrix synthesis. AB - The purpose of this study was to investigate factors which enhanced the compressive properties of alginate/chondrocyte constructs. Firstly, we studied the effect of biochemical composition (high, mid and low guluronic acid content) and sterilization method on alginate properties. Secondly, we studied the biosynthetic characteristics of chondrocytes in three different alginate compositions and performed mechanical tests to determine whether the synthesis of cartilage matrix components could significantly enhance the compressive properties. 2% alginate solutions containing an initial cell density of 4 x 10(6) cells/ml were cast into cylinders and cultured for seven weeks. Compression tests, biochemistry, immunohistochemistry and electron microscopy were performed at fixed intervals during the seven-week culture period. The dynamic modulus, peak strain, and peak stress were maximum for alginate with the highest guluronic acid content. The presence of cells and their respective matrix components enhanced the equilibrium modulus of the constructs for all types of alginate, though this effect was small. Alginate containing the middle amount of guluronic acid resulted in constructs which were both mechanically stable and which promoted synthesis of cartilage matrix proteins. In experiments and applications in which the mechanical integrity of the alginate is important, the composition and purity of the alginate and its method of sterilization should be selected with care. PMID- 11398866 TI - Hyaluronan suppressed nitric oxide production in the meniscus and synovium of rabbit osteoarthritis model. AB - Nitric oxide (NO) plays an important role in cartilage degeneration, and NO donors induce meniscus degeneration and synovium inflammation. This study evaluated the effect of intraarticular injections of hyaluronan (HA) on NO production in meniscus and synovium using an experimental osteoarthritis (OA) model. Thirty-six New Zealand white rabbits underwent unilateral anterior cruciate ligament transection (ACLT), and were divided into three groups. Four weeks after ACLT, the HA group started to receive intraarticular HA injections once a week for 5 weeks; the vehicle group started to receive the carrier of HA; and the no injection group, no treatment. All ACLT knees were harvested at the 9th week. Meniscus and synovium sections were examined by immunohistochemistry for nitrotyrosine. The pieces of these two tissues were cultured for 24 h. Culture supernatants were analyzed for nitrite concentration. The amount of NO produced by the meniscus was much larger than that produced by the synovium. NO productions in the meniscus and synovium of the HA group were significantly lower than those of the other groups. The results suggest that the inhibition of NO production in meniscus and synovium might be a part of the mechanism of the therapeutic effect of HA on OA. PMID- 11398867 TI - Cheap drugs? Part 1: a challenge to multinationals and to patent rights. PMID- 11398868 TI - Embryo (cells) for research. PMID- 11398869 TI - Cheap drugs? Part 2: WHO intervenes. PMID- 11398870 TI - Italy: colleagues will assure the pain does not return. PMID- 11398871 TI - Management of carcinoma of unknown primary site (CUP), any changes? PMID- 11398872 TI - What can we expect from liposomal drugs? PMID- 11398873 TI - Chemotherapy in elderly patients with colorectal cancer. AB - BACKGROUND: Colorectal cancer is usually diagnosed in patients around 70 years of age. With a continuous increase in life expectancy we may expect a higher number of elderly patients in the future. Because patients above 70 or 75 years are often excluded there is uncertainty as to what extent systemic adjuvant and palliative treatment should be offered to elderly patients. METHODS: We reviewed the available literature on adjuvant and metastatic colorectal cancer in order to identify reports on elderly patients treated within chemotherapy trials. RESULTS: Only about 20% of patients entering clinical trials belong to the age group of over 70 years and represent the minority of the very fit patients. Compared to their younger counterparts 5-FU-based treatment appears to be equally effective and more toxic according to some reports. Data regarding raltitrexed, oral fluoropyrimidines, topoisomerase I inhibitors or DACH-platin derivates are limited but suggest no age-specific differences in activity or toxicity. CONCLUSIONS: Elderly patients should not be excluded from clinical trials and studies in unfit elderly patients are warranted. Elderly patients need more attention regarding their functional, social and mental status. Fit elderly patients should be offered adjuvant or palliative chemotherapy. PMID- 11398874 TI - Palliative medicine and medical oncology. AB - Traditionally, medical oncology and palliative care have been considered two distinct and separate disciplines, both as regards treatment objectives and delivery times. Palliative care in terminal stages, aimed exclusively at evaluating and improving quality of life, followed antitumor therapies, which concentrated solely on quantitative results (cure, prolongation of life, tumoral mass shrinkage). Over the years, more modern concepts have developed on the subject. Medical oncology, dealing with the skills and strategic co-ordination of oncologic interventions from primary prevention to terminal phases, should also include assessment and treatment of patients' subjective needs. Anticancer therapies should be evaluated in terms of both the quantitative and qualititative impact on patients' lives. Hence, the traditional view of palliative care has to be modified: it constitutes a philosophical and methodological approach to be adopted from the early phases of illness. It is not the evident cultural necessity of integrating medical oncology with palliative medicine that may be a matter of argument, but rather the organizational models needed to put this combined care into practice: should continuous care be guaranteed by a single figure, the medical oncologist, or rather by an interdisciplinary providers' team, including full-time doctors well-equipped for palliative care? In this paper the needs of cancer patients and the part that a complete oncologist should play to deal with such difficult and far-reaching problems are firstly described. Then, as mild provocation, data and critical considerations on the ever increasing needs of palliative care, the present shortcomings in quality of life and pain assessment and management by medical oncologists, and the uncertain efficacy of interventional programmes to change clinical practice are described. Finally, a model of therapeutic continuity is presented. which in our view is realistic and feasible: an Oncologic Programme as the unifying process, and the Comprehensive Cancer Centre, or the Oncologic Department, the delivering structure. PMID- 11398875 TI - Prognostic factors and treatment effects on survival in acute myeloid leukemia of M6 subtype: a retrospective study of 54 cases. AB - classification system of acute myeloid leukemia (AML) which designates it as M6 AML. This report describes the data of 54 patients with newly diagnosed M6 AML, consecutively seen in our hospital between May 1976 and May 1999. PATIENTS AND METHODS: There were 40 males and 14 females. Median age was 59 years. Pancytopenia was the most common feature at diagnosis. Twenty-six percent of cases presented with secondary AML. Karyotype was successfully performed in 35 cases. Eleven patients presented with normal karyotype, nine with simple karyotypic abnormalities, and fifteen with major karyotypic abnormalities. Fifty of the fifty-four patients received one or two courses of induction chemotherapy combining anthracyclines with cytarabine according to different successive protocols. One elderly patient only received low-dose cytarabine, and three patients died before any chemotherapy could be given. RESULTS: Complete remission (CR) was achieved in 29 cases (54%, 95% confidence interval (CI): 40%-67%). As post-remission therapy, four patients could be allografted, and two underwent autologous transplantation. All other treated patients received continuation chemotherapy. Twenty-one patients have relapsed (72%). Median time to relapse was six months. Among those patients, only eight achieved a second CR (38%). The median disease-free survival (DFS) was eight months (95% CI: 4-10 months) with a five-year survival rate of 17%. Median overall survival (OS) was nine months (95% CI: 5-12 months) with a five-year survival rate of 13%. In univariate analysis, poor prognostic factors for DFS were secondary AML (P = 0.05) and initial platelet count <50 x 109/l (P = 0.02). Poor prognostic factors for OS were age > or = 60 years (P = 0.005), secondary AML (P = 0.05), initial 'blastic' fever (P = 0.0004), and initial haemoglobin level < 90 g/l (P = 0.03). All factors, but haemoglobin level, remained significant in the multivariate analysis. Although it was not statistically significant, there was a trent for a better prognosis of M6 patients presenting with normal karyotype as compared to those displaying chromosomal abnormality. CONCLUSIONS: This retrospective analysis points to a somewhat heterogenous group of AML in terms of clinical and biological features, and outcome. Distinctive subgroups can be identified according to prognostic factors related to survival. A larger multicenter study with well-defined diagnostic criteria is warranted to further clarify treatment effects. PMID- 11398876 TI - Liposomal daunorubicin in the treatment of relapsed or refractory non-Hodgkin's lymphoma. AB - PURPOSE: To assess the efficacy and toxicity of liposomal daunorubicin administered as a two-hour intravenous infusion to patients with relapsed or refractory non-Hodgkin's lymphoma (NHL). PATIENTS AND METHODS: Eligible patients had relapsed or refractory NHL with measurable or evaluable disease, and low grade, select intermediate grade, or mantle cell pathologic types. Prior exposure to an anthracycline or anthracenedione was allowed. Liposomal daunorubicin at a dose of 100 mg/m2 was given intravenously over a minimum of 120 minutes every 3 weeks. as a single agent. RESULTS: Thirty-three patients were accrued: twenty three (70%) had low-grade histologies; six (18%) had intermediate-grade histologies (follicular large-cell and diffuse small cleaved); and four (12%) patients had mantle-cell lymphoma. Eighteen (55%) had received two or more prior regimens; fourteen (42%) received a prior anthracycline. A median of six cycles of liposomal daunorubicin were administered (range 1-15). Of 31 patients evaluable for response, 2 complete and 10 partial remissions were documented for a major response rate of 39% (95% confidence interval (CI): 22%-58%). The median duration of response was 19.5 months (range 4.3-41.1+). Six responders (50%) had received a prior anthracycline; one responder had mantle-cell histology. The major toxicities were grade 3 or 4 neutropenia in 26 patients (79%), mild to moderate nausea in 22 (67%), and fatigue in 16 (48%). CONCLUSIONS: Liposomal daunorubicin at 100 mg/m2 every three weeks has activity in patients with relapsed or refractory NHL, including patients with prior exposure to an anthracycline. Further studies of liposomal daunorubicin in combination with other agents are warranted. PMID- 11398877 TI - A multicenter randomized clinical trial comparing paclitaxel-cisplatin-etoposide versus cisplatin-etoposide as first-line treatment in patients with small-cell lung cancer. AB - BACKGROUND: Previous phase I-II studies have shown that the combination of paclitaxel-cisplatin-etoposide (TEP) is very active and well tolerated in patients with small-cell lung cancer (SCLC). In order to compare the TEP combination to cisplatin etoposide (EP) regimen as front-line treatment in patients with SCLC, we conducted a randomised multicenter study. PATIENTS AND METHODS: One hundred thirty-three chemotherapy-naive patients with histologically proven limited or extensive stage SCLC were randomised to receive either paclitaxel 175 mg/m2 i.v. three-hour infusion on day 1 and cisplatin 80 mg/m2 i.v. on day 2 and etoposide 80 mg/m2 i.v. on days 2-4 with G-CSF support (5 mcg/kg s.c. days 5-15) or cisplatin 80 mg/m2 i.v. on day 1 and etoposide 120 mg/m2 i.v. on days 1-3 in cycles every twenty-eight days. RESULTS: Due to excessive toxicity and mortality observed in the TEP arm, an early interim analysis was performed and the study was closed. Sixty-two patients received two hundred sixty-one cycles of TEP and seventy-one patients three hundred twenty three cycles of EP The two patient groups were well balanced for age, sex, performance status, stage of disease and the presence of abnormal LDH at diagnosis. In an intention-to-treat overall analysis both regimens were equally active with a complete and partial response rate of 50% (95% confidence interval (CI): 37.5%-62.4%) for TEP and 48% (95%) CI: 36.2%-59.5%) for EP (P = 0.8). The median time to disease progression was 11 months for TEP and 9 months for EP (P = 0.02). The duration of response, one-year survival and overall survival were similar in the two arms. Similarly, in an intention-to-treat subgroup analysis of patients with limited or extensive stage disease, there was no difference in the activity between the two regimens except of a longer median time to disease progression in the extensive stage in favour of the TEP regimen, eight versus six months (P = 0.04). However, there were eight toxic deaths in the TEP arm versus none in the EP arm (P = 0.001). Moreover, the TEP regimen was associated with more severe toxicity than the EP regimen in terms of grade 4 neutropenia (P = 0.04), grade 3-4 thrombocytopenia (P = 0.02), febrile neutropenia (P = 0.08), grade 3-4 diarrhea (P = 0.01), grade 3-4 asthenia (P = 0.05) and grade 3 neurotoxicity (P = 0.06). CONCLUSIONS: In this early terminated study, the TEP regimen was significantly more toxic than the EP regimen. The TEP regimen is associated with significant toxicity and mortality, and should not be used outside of a protocol setting. For future investigations, dose and schedule modifications are necessary to reduce toxicity. PMID- 11398878 TI - Dose escalation of cytotoxic drugs using haematopoietic growth factors: a randomized trial to determine the magnitude of increase provided by GM-CSF. AB - BACKGROUND: The magnitude of chemotherapy dose escalation made possible by the use of recombinant haematopoietic growth factors has not been quantified in a randomized trial. PATIENTS AND METHODS: Patients with refractory or relapsing Hodgkin's disease were randomized to receive the Dexa-BEAM regimen with escalating etoposide doses supported by placebo or granulocyte-macrophage colony stimulating factor (GM-CSF). Using an adaptive sampling method independently in both arms, the etoposide dose was escalated until the maximal tolerated dose for the first cycle was reached. RESULTS: Thirty patients were randomized to GM-CSF and thirty to placebo. The etoposide dose could be escalated considerably in both treatment arms. Maximal etoposide dose for the first cycle was 1920 mg/m2 for patients receiving GM-CSF and 1160 mg/m2 for patients receiving placebo (P = 0.045 one-sided), corresponding to a 65% higher etoposide dose and a 13% higher dose intensity with GM-CSF. Dose-limiting events were similar in both arms, consisting mainly of prolonged neutropenia and consecutive infections. Treatment efficacy was not different in the two treatment groups. CONCLUSIONS: While GM-CSF permits a somewhat higher dose escalation than placebo, the increase in dose intensity provided by GM-CSF is small. The use of CSF for interval reduction rather than dose escalation is the more effective strategy for dose intensification. PMID- 11398879 TI - FDG-PET for detection of pulmonary metastases from malignant primary bone tumors: comparison with spiral CT. AB - BACKGROUND: The purpose was the comparison of positron emission tomography using F-18-fluorodeoxy-glucose (FDG-PET) and spiral thoracic CT to detect pulmonary metastases from malignant primary osseous tumors. PATIENTS AND METHODS: In 71 patients with histologically confirmed malignant primary bone tumors (32 osteosarcomas, 39 Ewing's sarcomas) 111 FDG-PET examinations were evaluated with regard to pulmonary/pleural metastases in comparison with spiral thoracic CT. Reference methods were the clinical follow-ups for 6-64 months (median 20 months) or a histopathologic analysis. RESULTS: In 16 patients (23%) reference methods revealed a pulmonary/pleural metastatic disease. FDG-PET had a sensitivity of 0.50, a specificity of 0.98, and an accuracy of 0.87 on a patient based analysis. Comparable values for spiral CT were 0.75, 1.00, and 0.94. It was shown that no patient who had a true positive FDG-PET had a false negative CT scan, nor was a pulmonary metastases detected earlier by FDG-PET than by spiral CT. CONCLUSIONS: There seems to be a superiority of spiral CT in the detection of pulmonary metastases from malignant primary bone tumors as compared with FDG-PET. Therefore, at present a negative FDG-PET cannot be recommended to exclude lung metastases. However, as specificity of FDG-PET is high, a positive FDG-PET result can be used to confirm abnormalities seen on thoracic CT scans as metastatic. PMID- 11398880 TI - E400P in advanced seminoma of good prognosis according to the international germ cell cancer collaborative group (IGCCCG) classification: the Spanish Germ Cell Cancer Group experience. AB - PURPOSE: To evaluate the efficacy and toxicity of primary chemotherapy with the schedule E400P in the treatment of patients with early stage II (IIa and IIb) and advanced seminoma of good prognosis according to the international classification (IGCCCG). PATIENTS AND METHODS: Sixty-four patients were included. E400P consisted of cisplatin 25 mg/m2/day and etoposide 100 mg/m2/day for four days, every three weeks. Royal Marsden stages were IIab: 53% and IIc-IV: 47%. Twenty three percent had high BHCG levels, twenty-seven percent had LDH > 2 x N. Sixty two patients were of good prognosis according to the Medical Research Council classification. RESULTS: Response rate was 98% (69% complete remission, 29% residual disease). After a median follow-up of 34 months, treatment failure was seen in 7 patients (11%). Neutropenia (32%) was the most relevant grade 3-4 toxicity. Other important grade 3-4 side effects were found in less than 5%. Three-year time to treatment failure (TTF) was 89% (95% confidence intervals (CI): 80%-97%) for all patients, 91% (95% CI: 80%-99%) for stages IIa-b, and 87% (95% CI: 74%-99%) for stages IIc-IV. Three-year overall survival (OS) was 97% (95% CI: 93%-99%) for all patients and 95% (95% CI: 85%-99%) for stages IIa-b. CONCLUSIONS: E400P was a very active and safe regimen in good-prognosis advanced seminoma, with low toxicity rates. Definitive comparisons of this regimen with radiotherapy in stages IIa-b or with the more standard E500P or BEP, could be of interest. PMID- 11398881 TI - Phase I-II study of pegylated liposomal cisplatin (SPI-077) in patients with inoperable head and neck cancer. AB - BACKGROUND: Concomitant chemoradiotherapy (CCRT) for squamous cancers of the head and neck (SCCHN) improves survival but increases toxicity. Pegylated liposomes localise to solid cancers and may deliver radiosensitizing agents preferentially to tumour tissue, potentially improving the therapeutic ratio of CCRT. PATIENTS AND METHODS: A phase I-II trial of pegylated liposome encapsulated cisplatin (SPI 077) was conducted in 18 patients with treatment-naive locally advanced, inoperable SCCHN. The first 10 patients received 2 cycles of 200 mg/m2, and the next 8 received 260 mg/m2, every 3 weeks before commencing radical radiotherapy (RT). RESULTS: Only 2 of 18 (11%) patients had partial responses to SPI-077 with 2 responses in 29 (6.9%) evaluable sites. SPI-077 was tolerated well with no haematological, renal, hepatic or neurological toxicities. Nausea and vomiting were minimal. There were no drug-related delays in the delivery of RT. RT-induced mucosal and cutaneous toxicity were not significantly increased. CONCLUSIONS: SPI 077 is essentially inactive against SCCHN and, in its present formulation, does not merit further evaluation as induction chemotherapy or as part of a CCRT approach. PMID- 11398882 TI - Phase II trial of liposomal daunorubicin in malignant pleural mesothelioma. AB - BACKGROUND: To assess the response rate, toxicity and survival in patients with malignant pleural mesothelioma treated with liposomal daunorubicin. The study design allowed for dose escalation pending toxicity. PATIENTS AND METHODS: Liposomal daunorubicin (DaunoXome, Nexstar, USA) 120 mg/m2 was administered every 21 days to a maximum of 6 cycles. Patients had to have histologically-proven malignant pleural mesothelioma. Patients were all chemotherapy-naive with ECOG performance status 0-2. RESULTS: Fourteen patients were enrolled. There were no objective or symptomatic responses though nine patients (64%) had stable disease on therapy. Myelosuppression was the major toxicity with 9 of 11 patients evaluable for toxicity experiencing grade 3 or 4 neutropenia. Other toxicities seen in at least 30% of patients included grade 3 infection and grade 2 nausea and vomiting. The median overall survival by intention-to-treat analysis was 6.1 months from the time of first treatment. The median duration of stable disease from time of first treatment for patients not progressing on therapy was 5.1 months. CONCLUSIONS: Liposomal daunorubicin 120 mg/m2 has no useful clinical activity in patients with malignant pleural mesothelioma. Toxicity was substantial with most patients experiencing at least one episode of grade 3 or 4 neutropenia. Liposomal daunorubicin cannot be recommended for patients with malignant pleural mesothelioma. PMID- 11398883 TI - A phase II trial of irinotecan (CPT-11) for unresectable biliary tree carcinoma. AB - BACKGROUND: Unresectable adenocarcinomas of the biliary tree have a very poor prognosis. No good chemotherapeutic regimen is available. Irinotecan has not yet been fully tested in this disease. We evaluated its activity in unresectable bile duct cancers. PATIENTS AND METHODS: Twenty-five consecutive eligible patients at our two institutions were treated with irinotecan at a starting dose of 125 mg/m2. A cycle consisted of once-a-week treatments for four consecutive weeks, followed by two weeks of rest. All patients were required to have histologically confirmed diagnosis, clinically documented metastatic or unresectable carcinoma and measurable disease. Patients were evaluated for response, toxicity, and survival. RESULTS: A total of 83 cycles of therapy were delivered. Two patients had a partial response (8%; 95% confidence interval (CI): 0%-18%) and ten additional patients had stable disease for at least two months (40%; 95% CI: 20.8%-59.2%). The therapy was well tolerated, with moderate myelosuppression and diarrhea as the main toxicities. The overall median survival was 10 months. CONCLUSIONS: Irinotecan has minimal activity in biliary tree carcinomas, but is well tolerated with appropriate supportive care, and produces occasional objective responses. PMID- 11398884 TI - Intensified chemotherapy supported by DMSO-free peripheral blood progenitor cells in breast cancer patients. AB - BACKGROUND: The majority of high-dose chemotherapy (HDC)-related complications results from bone marrow aplasia, but the graft infusion per se may cause adverse reactions due to the injection of both dimethyl sulfoxide (DMSO) and cell lysis products. We evaluated the feasibility of a two-step chemotherapy regimen with peripheral blood progenitor cell (PBPC) support in association with a novel procedure to remove DMSO and products of cell lysis from the cryopreserved cells. PATIENTS AND METHODS: Stage III and IV breast cancer patients received induction chemotherapy with three cycles of CEF (cyclophosphamide 600 mg/m2, epirubicin 100 mg/m2, 5-fluorouracil 600 mg/m2) followed by three cycles of HDC consisting of escalating doses of cyclophosphamide (dose range 1200 3000 mg/m2) and carboplatin (dose range 600-1000 mg/m2), supported by DMSO-free PBPC reinfusion. DMSO was removed by a washing/enzymatic digestion procedure. RESULTS: Twenty patients received induction chemotherapy and eighteen completed the entire chemotherapy program; a total of fifty-four cycles of HDC were administered. Dose limiting toxicity of HDC was long-lasting grade 4 neutropenia associated with documented infection. The maximum tolerated dose (MTD) was cyclophosphamide 3000 mg/m2 and carboplatin 600 mg/m2. No side effects related to PBPC reinfusion were observed. CONCLUSIONS: The proposed two-step chemotherapy regimen, associated with a novel washing/enzymatic digestion procedure, is feasible in advanced breast cancer patients in the absence of complications related to the specific toxicity of PBPC reinfusion. PMID- 11398885 TI - Activity of the dolastatin analogue, LU103793, in malignant melanoma. AB - LU103793, a synthetic analogue of dolastatin 15, showed interesting pre-clinical activity in melanoma xenografts. In this phase II multicentre trial, 80 chemotherapy-naive patients with metastatic melanoma received a total of 218 cycles of treatment. The response rate showed one complete and three partial responses of median duration six months (range 3-9.1). Toxicity was moderate, mostly haematological (neutropenia grade 4 in 16%, grade 3 in 3%). There were no significant problems with hypertension or other non-haematological toxicities. PMID- 11398886 TI - Clinical outcome after front-line intensive sequential chemotherapy (ISC) in patients with aggressive non-Hodgkin's lymphoma and high-risk international prognostic index (IPI 3): final analysis of survival in two consecutive ISC trials. AB - BACKGROUND: Aggressive non-Hodgkin's lymphomas (NHL) in patients under the age of 60 have a very poor prognosis when the international prognostic index (IPI) is high, with an age-adjusted (Aa)-IPI score at 3. In such patients, conventional chemotherapy results in a low complete response (CR) rate of 46%, a five-year survival and disease-free survival (DFS) of 32% and 58%, respectively. For this report we have analyzed whether front-line high-dose chemotherapy could influence the outcome of this group of patients. PATIENTS AND METHODS: From 1992 onwards we conducted two pilot clinical trials of intensive sequential chemotherapy (ISC) with growth factors and blood stem cell support as initial treatment in 62 poor risk patients with aggressive NHL. Of these patients, 33 were considered to be a high-risk group based on the Aa-IPI. RESULTS: The median age was 42 years (range 21-60). The treatment was completed in 88% of patients, 86% receiving greater than 75% or more of the projected dose-intensity. Twenty patients (61%) achieved a CR. At a median follow-up of 48 months (range 26-86), the estimated five-year survival and DFS was 51% (95% confidence interval (CI): 34%-68%) and 70% (95% CI: 50%-90%), respectively. CONCLUSION: These results suggest that primary treatment using high-dose therapy supported by both growth factors and peripheral blood stem cells can cure up to 50% of high-risk patients with malignant lymphomas. PMID- 11398887 TI - Oxaliplatin and protracted continuous 5-fluorouracil infusion in patients with pretreated advanced colorectal carcinoma. AB - BACKGROUND: Both OHP and 5-FU are clinically active as single agents in the treatment of metastatic colorectal cancer (MCRC). Clinical and laboratory studies suggest a synergistic interaction between these agents. This phase II study was performed to evaluate the activity of a schedule including OHP and protracted 5 FU infusion in 5-FU-resistant MCRC. PATIENTS AND METHODS: From October 1997 to January 2000, 50 patients with measurable progressive MCRC after one or more 5-FU based regimens were treated. OHP (2-3-hour i.v. infusion) on day 1 and 5-FU (protracted i.v. infusion using elastomeric/electronic pump through a central venous catheter) on days 1-21 were administered every 3 weeks, at the following 4 dose levels: 1) OHP 100 mg/m2 + 5-FU 200 mg/m2 (21 patients); 2) OHP 100 mg/m2 + 5-FU 250 mg/m2 (3 patients); 3) OHP 130 mg/m2 + 5-FU 200 mg/m2 (10 patients); 4) OHP 130 mg/m2 + 5-FU 250 mg/m2 (6 patients). RESULTS: Objective responses were 1 (2%) CR; 10 (20%) PR, for a median duration of 8 months; 23 (46%) stable diseases, for a median duration of 6 months: 16 (32%) progressions. CR + PR was higher in patients who had previously received no more than one line of chemotherapy for metastatic disease as compared with patients who had received two or more lines of therapy (33% vs. 5%, P < 0.01). The median time to progression was four months (one to nine). All dose levels (313 cycles) were well tolerated with mild toxicity. Major toxicity (grade 3 WHO) included: anaemia in 1 patient (2%), nausea and vomiting in 1 patient (2%), diarrhoea in 4 patients (8%) and stomatitis in 1 patient (2%); grade I and 2 peripheral neuropathy were encountered, respectively, in 30 (60%) and 8 (16%) patients. The median survival was 13 months (9-17), with 32 patients still alive after a median follow-up of 8 months. CONCLUSIONS: This study suggests that 1) OHP plus protracted 5-FU infusion is an active combination in MCRC patients resistant to pre-treatment bolus 5-FU; 2) it has a good tolerability profile and 3) the optimum dose level is OHP 130 mg/m2 and 5-FU 250 mg/m2. PMID- 11398888 TI - C-erbB-2 oncoprotein overexpression identifies a subgroup of estrogen receptor positive (ER+) breast cancer patients with poor prognosis. AB - PURPOSE: To investigate the predictive value of c-erbB-2 oncoprotein expression as compared with established histopathological and cytometric indicators of disease evolution in breast carcinoma. PATIENTS AND METHODS: A short-term retrospective study was conducted on a series of 306 breast cancer patients. Classic prognostic factors included tumour size, nodal involvement, histological grading, and hormone receptor status. Flow cytometric DNA ploidy and S-phase fraction (SPF) were also assessed. A Cox proportional hazards regression model was used for multivariate statistical analysis. RESULTS: c-erbB-2 overexpression was present in 43 out of 295 (14.6%) tumours, and showed a statistically significant correlation with high histological grade, DNA aneuploidy, high SPF and lack of estrogen receptors (ER). Univariate analysis revealed its association with worse disease-free survival (DFS) and overall survival (OS). The combined evaluation of c-erbB-2 with ploidy and SPF defines a variable (P + S + c) that showed a significant correlation with disease outcome. By multivariate analysis, only nodal status (P < 0.001) and P + S + c subgrouping (group 2: P = 0.002; group 3: P = 0.001) in relation to DFS, and nodal status (P = 0.001) and DNA ploidy (P = 0.006) in relation to OS, retained independent prognostic significance. Subset analyses showed that cytometric parameters, P + S + c subgrouping and hormone receptors were significantly correlated with disease outcome in node-positive patients, whereas in node-negative subgroup no prognostic indicators were found. c-erbB-2 overexpression exhibited a trend in node-positive breast cancer (DFS: P = 0.068; OS: P = 0.086), and significant correlation with poor clinical evolution in ER positive patients (DFS: P = 0.015; OS: P = 0.004), mostly receiving tamoxifen. CONCLUSIONS: c-erbB-2 is an independent prognostic indicator of DFS when evaluated in conjunction with ploidy and SPE. It also seems to predict response to tamoxifen therapy, by identifying a subgroup of ER positive (ER+) breast cancer patients with poor prognosis. PMID- 11398889 TI - Carcinoma of an unknown primary site: a chemotherapy strategy based on histological differentiation--results of a prospective study. AB - BACKGROUND: To evaluate the efficacy and toxicity of a chemotherapy strategy based on histological differentiation, for patients with carcinoma of unknown primary site. PATIENTS AND METHODS: Forty-eight patients were prospectively included in the trial. Thirty patients with poorly-differentiated carcinoma or poorly-differentiated adenocarcinoma (group A) received a combination of cisplatin and etoposide. Patients with a responsive or stable disease after two cycles received the same regimen plus bleomycin, ifosfamide and G-CSF. Eighteen patients with well- or moderately-differentiated carcinoma (group B) received cisplatin, continuous infusion 5-fluorouracil (5-FU) and alpha-interferon. Treatment was maintained in case of response or stable disease for up to six cycles. RESULTS: The overall response rate (RR) for the entire group is 43% (95% confidence interval (CI): 35.9%-50.1%): seven CR and five PR in group A (RR = 40%) and six CR and two PR in group B (RR = 44%). Grade 4 leucopenia was observed in 22 (46%) patients and sepsis in 3 (6%). Median survival is 9.4 months (range 5 13.7 months) and 16.1 months (range 11.8 20.3 months), respectively. CONCLUSIONS: This chemotherapy strategy is one way to achieve high response rates, particularly for patients with well- or moderately-differentiated adenocarcinoma usually considered poorly chemosensitive. PMID- 11398890 TI - A phase II study of gemcitabine and cisplatin combination as induction chemotherapy for untreated locally advanced cervical carcinoma. AB - BACKGROUND: Cisplatin-based chemoradiation for locally advanced cervical carcinoma is now the standard of care for most patients with cervical carcinoma. However, induction chemotherapy followed by surgery, particularly with newer agents or combinations remains to be explored. This study was undertaken to evaluate the antitumor activity and toxicity of gemcitabine in combination with cisplatin for untreated locally advanced cervical carcinoma. PATIENTS AND METHODS: Open-label, single center, phase II, non-randomized study of neoadjuvant gemcitabine plus cisplatin. Forty-one patients with histologic diagnosis of cervical carcinoma, with no previous treatment and staged as IB2 to IIIB, were treated with three 21-day courses of cisplatin 100 mg/m2 day I and gemcitabine 1000 mg/m2 days 1 and 8, followed by locoregional treatment with either surgery or concomitant chemoradiation. Response and toxicity were evaluated before each course and at the end of chemotherapy. RESULTS: All patients were evaluated for toxicity and 40 for response. The overall objective response rate was 95% (95% confidence interval (CI): 88%-100%) being complete in 3 patients (7.5%) and partial in 35 (87.5%). A complete pathological response was found in 6 (26%) of the 23 patients that underwent surgery. Granulocytopenia grades 3-4 occurred in 13.8% and 3.4% of the courses, respectively, whereas non-hematological toxicity was mild. CONCLUSIONS: Induction chemotherapy with the combination of gemcitabine and cisplatin is highly active for untreated cervical cancer patients and has an acceptable toxicity profile. PMID- 11398891 TI - Phase I dose escalation study of topotecan combined with alternating schedules of paclitaxel and carboplatin in advanced solid tumors. AB - BACKGROUND: Combining topotecan with other cytotoxics has been problematic due to marrow suppression. A phase I trial was initiated to identify the optimal sequence and maximum-tolerated dose of topotecan in combination with paclitaxel and carboplatin. PATIENTS AND METHODS: Patients with advanced cancer and performance status ECOG < or = 2. The starting dose was paclitaxel 175 mg/m2 day 1, carboplatin AUC 6.0 day 1, and topotecan 0.5 mg/m2 daily day 1-5 (early sequence). The next course of paclitaxel and carboplatin administration was delayed to day 5 (late sequence). Treatment was repeated every three weeks. After determining maximum-tolerated dose without cytokines, granulocyte colony stimulating factor (G-CSF) was added and further dose escalation was pursued. RESULTS: Fifty-one patients were entered; men: women ratio 30:21. Dose-limiting toxicity (DLT) for the early sequence was neutropenia at doses paclitaxel mg/m2/carboplatin AUC 5/topotecan mg/m2 (PCT) 175/5/0.75 for four to five days. DLT for the late sequence was neutropenia at PCT doses of 175/5/ 1.0 for four days. G-CSF 5 microg/kg subcutaneously starting day 6 permitted further topotecan dose escalation. After adding G-CSF, late sequence DLT was neutropenia at doses 175/5/1.25 for four days. Forty-six patients were evaluable for response and of those, there were thirteen partial responses. CONCLUSIONS: The late sequence resulted in less toxicity and was better tolerated. The early sequence maximum tolerated dose (MTD) was 175/6/0.5 for five days. The late sequence MTD was PCT 175/5/0.75 for five days. The late sequence MTD with G-CSF was 175/5/1.0 for four days. The recommended phase II PCT dose is the late sequence 175/5/1.0 for four days with G-CSF. PMID- 11398892 TI - Single-agent gemcitabine in patients with resistant small-cell lung cancer. AB - OBJECTIVE: This study was conducted to assess the activity and toxicity ofgemcitabine in patients with resistant small-cell lung cancer (SCLC). PATIENTS TAND METHODS: Forty-one patients with limited- or extensive-stage SCLC, who were previously treated with at least one chemotherapeutic regimen and progressed during or within three months of finishing the last regimen, were treated with 1000 mg/m2 gemcitabine on days 1, 8, and 15 of a four-week cycle. RESULTS: Thirty eight patients were evaluable for response. Five partial and no complete responses were seen, for an overall response rate of 13% (95% confidence interval (CI): 6%-27%). Time to progression varied from 4 to 20 weeks, with a median of 8 weeks. Median survival was 17 weeks (range 4-84 weeks). Hematological toxicity mainly consisted of NCI-CTC grade 3 thrombocytopenia (29% of patients) and, to a lesser extent, grade 3 leukopenia (18% of patients). Non-hematological toxicity was mild, with nausea being the most commonly reported event. CONCLUSIONS: Gemcitabine has modest activity in patients with resistant SCLC. There is some non-cross resistance to most agents against SCLC. PMID- 11398893 TI - Primary papillary serous carcinoma of the peritoneum in a man. AB - BACKGROUND: Primary papillary serous carcinoma of the peritoneum is a well-known entity in women. The tumour is derived from the extraovarian mesothelium and the pelvis and lower abdomen mesothelia. The treatment strategies are similar to ovarian serous papillary carcinoma. PATIENTS AND METHODS: A case of primary serous papillary carcinoma of the peritoneum in a man is presented. The patient, 53 years old, died 2 months after diagnosis. RESULTS: The histologic and immunohistochemical studies of the tumour will be presented. These studies, made during lifetime and at autopsy of the patient, confirm a diagnosis of primary serous papillary carcinoma of the peritoneum. CONCLUSIONS: primary serous papillary carcinoma of the peritoneum can occur in men, and should be considered in the differential diagnosis in cases of abdominal carcinomatosis of unknown origin. Treatment options remain to be determined. PMID- 11398894 TI - Safe administration of irinotecan, oxaliplatin and raltitrexed in a DPD-deficient patient with metastatic colon cancer. AB - Dihydropyrimidine dehydrogenase deficiency is diagnosed more frequently and is now generally accepted as a potentially life-threatening condition. It predisposes patients receiving treatment with fluoropyrimidines such as 5 fluorouracil (5-FU) to severe and, in case of complete dihydropyrimidine dehydrogenase deficiency, often fatal toxicity. A patient who had severe side effects following standard dose adjuvant 5-FU exposure was diagnosed of having hereditary partial dihydropyrimidine dehydrogenase deficiency. When the patient relapsed with liver metastases, we treated him with the non-fluoropyrimidine cytotoxic agents irinotecan, oxaliplatin and raltitrexed in sequential manner, and were able to show that these drugs can be safely applied in patients with this metabolic defect. PMID- 11398895 TI - Geographical disparities in self-reported use of mammography and breast self examination according to the Swiss Health Survey. PMID- 11398896 TI - The role of mitomycin C in the treatment of patients with advanced colorectal cancer resistant to 5-fluorouracil-folinic acid chemotherapy. PMID- 11398897 TI - Mediators and intracellular mechanisms of NANC relaxation of smooth muscle in the gastrointestinal tract. PMID- 11398898 TI - Alpha-adrenoceptor agonists produce Ca2+ oscillations in isolated rat aorta: role of protein kinase C. AB - We investigated the relationship between tension development and the cytosolic free Ca2+ level ([Ca2+]i) in responses to norepinephrine (NE) and selective alpha2-adrenoceptor agonist, UK14,304 of the endothelium-denuded rat aorta loaded with fura PE-3. NE (3 x 10(-8) M) evoked a rapid increase in [Ca2+]i followed by slight decreasing to a steady state level and produced a contraction. After the NE-induced increase in [Ca2+]i had reached a maximum, the [Ca2+]i showed persistent oscillations. The Ca2+ oscillations were superimposed on the sustained increase in [Ca2+]i. UK14,304 (3 x 10(-6) M) also evoked an increase in [Ca2+]i and produced a contraction. However, the UK14,304-induced effect on [Ca2+]i was characterized by pronounced oscillations, and the amplitude of the sustained increase in [Ca2+]i was less than that seen with NE. Protein kinase C inhibitor, Ro31-8220 (3 x 10(-6) M) and verapamil (10(-5) M) abolished both NE and UK14,304 evoked Ca2+ oscillations. UK14,304-induced contractions were also strongly inhibited by Ro31-8220 and verapamil. However, NE induced contractions were partly inhibited by these inhibitors. The sustained increases in [Ca2+]i evoked NE and UK14,304 were not significantly inhibited by Ro31-8220 and verapamil. These results suggest that NE and UK14,304 produce Ca2+ oscillations during sustained contractions in rat aorta. The alpha2 adrenoceptor agonist, UK14,304 induced sustained contraction and Ca2+ oscillations may be due to PKC activation and opening of voltage-dependent L type Ca2+ channels. PMID- 11398899 TI - Influence of environmental, structural, and behavioral factors on the presence of phosphine in worker areas during fumigations in grain elevators. AB - Data-logging gas monitors with electrochemical cells sensitive to phosphine (PH3) were used to characterize concentrations of this common grain fumigant in and around grain elevators during fumigations. Twenty-four grain fumigations were observed, and each was monitored over a 5- to 8-day period. Phosphine gas, generated from aluminum phosphide fumigant applied to the grain, generally moved upward toward the grain surface and exited the bin at bin-top openings to the outside air or to enclosed worker areas. The upward air currents appeared to be the result of chimney effects, e.g., pressure differences resulting from buoyant air inside the warm grain and cooler, denser, ambient air. Significant wind effects on the PH3 concentration were also observed in the air between the grain surface and the bin roof. In enclosed areas located at the bin-top level, monitors located near the fill port or the fumigant dispenser recorded PH3 concentrations in excess of the exposure limit of 0.3 parts per million (ppm) about 35% of the time during grain fumigations. Phosphine concentrations between 0.31 and 1.0 ppm were observed 17.3% of the time, and concentrations in the ranges of 1.01-3.0, 3.01-10.0, and >10 ppm constituted 11.8%, 5.5%, and 0.3% of all readings, respectively, in bin-top worker areas. The likelihood of recording PH3 concentrations >0.3 ppm depended on ventilation practices. Fans in tunnels and open windows at aboveground locations appeared to greatly reduce the likelihood of high PH3 concentrations in enclosed areas. PMID- 11398900 TI - Proper use and improper images: the double-edged sword of the extra seat for farm machinery. PMID- 11398901 TI - Monitoring acetylcholinesterase levels in migrant agricultural workers and their children using a portable test kit. AB - The EQM Research, Inc., portable test kit was evaluated as a surveillance tool for blood cholinesterase levels among migrant workers and their children. Laboratory validation demonstrated a linear relationship between the reference Ellman and kit methods (Ellman = 0.95 x kit result + 0.82, r2 = 0.98). Pre- and post-season cholinesterase levels measured in 70 farm workers were within normal ranges, but significantly different at 28.5 and 29.7 U/g Hb, respectively (paired t-test, p = 0.014). Results from 98 migrant farm worker children and a comparison group of 53 age-matched non-agricultural children showed that cholinesterase levels were not significantly different between the agricultural and non agricultural children (ANOVA, p = 0.69). These data demonstrate that a portable test kit can provide useful data pesticide exposures when measurements are made in a temperature-controlled setting. PMID- 11398902 TI - Performance of an automatically deployable ROPS on ASAE tests. AB - In the U.S., approximately 132 agricultural tractor overturn fatalities occur per year. The use of rollover protective structures (ROPS), along with seat belts, is the best-known method for preventing these fatalities. However, one impediment to ROPS use is low-clearance situations, such as orchards and animal confinement buildings. To address the need for ROPS that are easily adapted to low-clearance situations, the Division of Safety Research, National Institute for Occupational Safety and Health, developed a prototype automatically deploying, telescoping ROPS (AutoROPS). The NIOSH AutoROPS consists of two subsystems. The first is a retractable ROPS that is normally latched in its lowered position for day-to-day use. The second subsystem is a sensor that monitors the operating angle of the tractor. If an overturn condition is detected by the sensor, the retracted ROPS will deploy and lock in the full upright position before ground contact. Static load testing and field upset tests of the NIOSH AutoROPS have been conducted in accordance with SAE standard J2194. Additionally, timed trials of the AutoROPS deployment mechanism were completed. The results of these tests show that the NIOSH AutoROPS has significant potential to overcome the limitations of current ROPS designs for use in low clearance as well as unrestricted clearance operations. PMID- 11398903 TI - The theory of planned behavior: use and application in targeting agricultural safety and health interventions. AB - Traditional farm safety programs rely mostly on iterations of knowledge-based components, typically lack local input into identification of issues and concerns, and are difficult to evaluate. Psychological models have been looked at as tools to address these problems. The Theory of Planned Behavior (ToPB) is a psychological model that has been used to understand behavioral beliefs and to provide a framework for using those beliefs as intervention targets. The fundamental characteristics of ToPB are reviewed. Examples of specifically stated behavioral questions are provided from ToPB application in field settings. Pertinent findings from an example study on personal protective equipment use and beliefs are summarized to illustrate advantages and disadvantages of using ToPB. ToPB can provide information useful in targeting interventions aimed at locally identified safety and health concerns. The use of beliefs identified using ToPB as intervention targets can increase the use of personal protective equipment beyond levels used prior to the intervention. And, ToPB has shown multiple correlations of 0.28 (up to 0.35 in other examples not reported) between intentions and self-reported behaviors, indicating that other contributory factors are present and need to be studied. PMID- 11398904 TI - Aminoglycoside adaptive resistance: importance for effective dosage regimens. AB - There are various pharmacodynamic features of the aminoglycosides that are thought to contribute to the benefits of once-daily administration, of which the ability to induce adaptive resistance is the least understood and discussed. However, this may be the most important characteristic conferring increased efficacy with extended interval dose administration. Adaptive resistance describes a reversible refractoriness to the bactericidal effect of an antibacterial agent. It is well documented for the aminoglycosides but has also been seen with the quinolones. It does not appear to be caused by a genetic mutational change but rather by a protective phenotypic alteration in bacterial characteristics. This includes reversible down-regulation of the active transport of aminoglycosides into gram-negative bacteria. In vitro, animal and clinical studies have shown that marked adaptive resistance of gram-negative bacteria to aminoglycosides occurs within 1-2 hours of the first dose. The duration of adaptive resistance relates directly to the half-life of elimination of the aminoglycoside. With normal human aminoglycoside pharmacokinetics, the resistance may be maximal for up to 16 hours after a single dose of aminoglycoside, followed by partial return of bacterial susceptibility at 24 hours and complete recovery at around 40 hours. With conventional dosage regimens, second and subsequent doses of aminoglycoside are given at the time of maximal resistance and this practice is also likely to reinforce the resistance. Dose administration at 24 hour intervals, or longer, may increase efficacy by allowing time for adaptive resistance to reverse. PMID- 11398905 TI - Farnesyltransferase inhibitors: potential role in the treatment of cancer. AB - New targets for drug discovery have been identified rapidly as a result of the many recent and rapid advances in the understanding of signal transduction pathways that contribute to oncogenesis. In particular, oncogenic Ras proteins have been seen as an important target for novel anti-cancer drugs. Since the decade-old identification and cloning of farnesyltransferase (FTase), a critical enzyme that post-translationally modifies Ras and other farnesylated proteins, FTase inhibitors (FTIs) have been under intense investigation designed to bring them to clinical practice for cancer therapy. FTIs can inhibit the growth of tumour cells in culture and in animal models, and are now in clinical trials. Interestingly, their mechanism of action is not as simple as originally envisioned, and Ras is probably not the most important farnesylated protein whose modification is inhibited as a result of FTI treatment. Although K-Ras can escape inhibition of processing by FTIs, tumours with oncogenically mutated K-Ras proteins can still be inhibited by FTI treatment. Indeed, Ras mutation status does not correlate with FTI sensitivity or resistance. Instead, it now appears likely that inhibition of the processing of other farnesylated proteins such as RhoB and the centromere-binding proteins CENP-E and CENP-F can explain the ability of FTIs to cause cell cycle arrest and apoptosis in preclinical studies, and even to cause regression in animal tumour models. Preclinical studies suggest the likelihood that FTIs will be useful in combination therapies with conventional treatment modalities including cytotoxics (especially paclitaxel) and radiation. Phase I combination trials are underway, and early phase II/III trials using FTIs as monotherapy are open for patients with a wide variety of cancers. Early preclinical results also suggest the possibility of using FTIs as chemopreventive agents. Studies to be completed over the next 2 or 3 years should define the appropriate patient populations, administration and scheduling necessary to optimise the use of these novel anticancer agents. PMID- 11398906 TI - Renoprotective therapy in patients with nondiabetic nephropathies. AB - End-stage renal failure (ESRF) represents a major health problem. Early diagnosis and effective measures to slow or to stop renal damage are essential goals for nephrologists to prevent or delay progression to ESRF. Identifying mechanisms of progressive parenchymal injury is instrumental in developing renoprotective strategies. Protein traffic through the glomerular barrier is an important determinant of progression in chronic nephropathies and proteinuria is the best predictor of renal outcome. At the moment, ACE inhibition is the most effective treatment in patients with chronic nondiabetic proteinuric nephropathies, reducing protein traffic, urinary protein excretion rate and progression to ESRF more effectively than conventional treatment. Low sodium diet and/or diuretic treatment may help to increase the antiproteinuric effect of ACE inhibitors by maximally activating the renin-angiotensin system. Intensified blood pressure control, whatever treatment is employed, also enhances the antiproteinuric response to ACE inhibitors. However, since this is not always sufficient to normalise urinary proteins and fully prevent renal damage, additional treatments may be needed in patients poorly or not responding to ACE inhibitors. These may include angiotensin II receptor antagonists, non-dihydropyridine calcium antagonists and perhaps low doses of nonsteroidal anti-inflammatory drugs. Preliminary data on multidrug treatments including these additional antiproteinuric agents are encouraging, but additional studies in larger patient numbers are needed to better define the risk/benefit profile of this innovative approach. PMID- 11398907 TI - Fluoroquinolones: place in ocular therapy. AB - The fluoroquinolones have become widely used antibacterial agents in the treatment of ocular infections, with topical, intravitreal and systemic routes of administration being used. In general, fluoroquinolones (such as ciprofloxacin, ofloxacin, lomefloxacin and norfloxacin) have good activity against gram-negative and gram-positive bacteria. Therapeutic concentrations are achieved in the cornea after topical administration so that the fluoroqinolones have largely replaced combination therapy for the treatment of bacterial keratitis. However, a second line agent is needed when resistance is likely, such as in disease caused by streptococcal species. Reversal of resistance to quinolones may not occur with withdrawal of the antibacterial. This stresses the importance of prudent prescribing to reduce the occurrence of resistance to quinolones. When used in therapeutic topical dosages, corneal toxicity does not occur. Similarly, retinal toxicity is not seen when fluoroquinolones are used at therapeutic dosages, systemically or topically. Corneal precipitation occurs, particularly with ciprofloxacin and to a lesser extent norfloxacin, but does not appear to interfere with healing. In the treatment of endophthalmitis there is reasonable penetration of systemic fluoroquinolones into the vitreous but sufficiently high concentrations to reach the minimum inhibitory concentration for 90% of isolates (MIC90) of all important micro-organisms may not be guaranteed. Systemic administration may be useful for prophylaxis after ocular trauma. PMID- 11398908 TI - Management of antimicrobial use in the intensive care unit. AB - Indications for the use of antimicrobials in critically ill patients are similar to those for other hospitalised patients. However, the selection of agents depends on the particular characteristics of patients in the intensive care unit (ICU), the form of presentation of infection, the type of infection and the bacteriological features of the causative pathogens. The use of antimicrobials in patients admitted to medical-surgical ICUs varies between 33 and 53%. The selection of empirical antimicrobials to be included in treatment protocols of the most common infections depends on the strong interrelationship between patient characteristics, predominant pathogens in each focus. and antimicrobials used for treatment. Epidemiological studies carried out in the past have identified the microorganisms most frequently responsible for community-acquired and nosocomial infections in patients admitted to ICUs. Susceptibility to antimicrobial agents may be different between each geographical area, between each hospital and even within the same hospital service. In addition, susceptibility patterns may change temporarily in relation to the use of particular antimicrobials or in association with other unknown factors so that assessment of endemic antimicrobial resistance patterns is very useful in order to tailor the antimicrobial regimens of therapeutic protocols. Antimicrobial use should not be a routine procedure. The clinical course of the patient (an indicator of effectiveness) should be closely monitored as well as the possible appearance of adverse effects and/or multiresistant pathogens. Controls are based on the assessment of plasma drug concentrations and microbiological surveillance to detect the presence of multiresistant strains or new antibacterial-resistant pathogens. Prevention of the development of multiresistant pathogens is the main goal of the ICU antimicrobial policy. Although a series of general strategies to reduce the presence of multiresistant pathogens have been proposed, the implementation of these recommendations in ICUs requires the cooperation of a member of the intensive care team. PMID- 11398909 TI - Thalidomide in gastrointestinal disorders. AB - Thalidomide was originally marketed as a sedative, but was removed from the market in 1961 after it was associated with an epidemic of severe birth defects. Subsequently, it has been shown to have therapeutic efficacy in a number of the gastrointestinal tract conditions characterised by immune dysregulation. The exact mechanism of the immunosuppressive effects of thalidomide is unknown; proposed mechanisms include inhibition of tumour necrosis factor alpha release and inhibition of angiogenesis. In chronic graft versus host disease, use of high dose thalidomide (1200 mg/day) may bring about a response in 20% of patients with refractory disease. Thalidomide 200 mg/day helps eradicate ulcers in 50% of patients with HIV-associated oral aphthous ulceration. In Behcet's disease, thalidomide 100 to 300 mg/day can decrease the number of mucocutaneous ulcers, although full remission occurs in less than 20% of patients. In Crohn's disease, thalidomide 50 to 300 mg/day may decrease the severity of mucosal disease and prompt closure of fistulae. Patients to be placed on thalidomide therapy must practice either abstinence or strict birth control; women must undergo regular pregnancy testing and utilise 2 forms of contraception. Other adverse effects include sedation (present in nearly all patients), symptomatic neuropathy (present in approximately 20%), and skin rashes. Given the potential toxicity, thalidomide use should generally be limited to clinical protocols with institutional review board oversight. PMID- 11398910 TI - Desloratadine. AB - Desloratadine is the orally active major metabolite of the nonsedating H1 antihistamine loratadine. The drug had no adverse cardiovascular effects in various animal models or when administered at 9 times the recommended adult dosage for 10 days in volunteers. Therapeutic dosages had no effects on wakefulness or psychomotor performance in healthy volunteers. No clinically significant interactions have been reported between desloratadine and drugs that inhibit the cytochrome P450 system, nor does the drug potentiate the adverse psychomotor effects of alcohol. Oral desloratadine 5 mg once daily for up to 4 weeks in patients with seasonal allergic rhinitis (SAR) significantly reduced nasal (including congestion) and non-nasal symptoms and improved health-related quality of life compared with placebo. Similar beneficial effects were observed in patients with SAR and coexisting asthma (in whom asthma symptoms and use of beta2-agonists were reduced). Desloratadine 5 mg once daily for 6 weeks significantly improved pruritus and reduced the number of hives compared with placebo in patients with chronic idiopathic urticaria (CIU). Sleep and daytime performance also improved. Desloratadine was well tolerated in clinical trials and had an adverse event profile similar to that of placebo in patients with SAR (with or without asthma) or CIU. PMID- 11398911 TI - Zoledronic acid. AB - Zoledronic acid (zoledronate) is a new generation bisphosphonate that inhibits osteoclast bone resorption. It was much more potent than other bisphosphonates at inhibiting 1,25-dihydroxyvitamin D3-induced hypercalcaemia in a rat model and calcium release in vitro. A single 5-minute intravenous infusion of zoledronic acid (4 or 8 mg) was significantly more effective than a 2-hour infusion of pamidronic acid (pamidronic acid disodium, pamidronate disodium) [90 mg] in normalising serum calcium levels in patients with hypercalcaemia of malignancy and resulted in a significantly longer median time to relapse (pooled analysis from 2 randomised, double-blind, parallel-group trials). There were no differences in tolerability between zoledronic acid and pamidronic acid in comparative trials; the most common events in pivotal trials were fever, anaemia, nausea, constipation and dyspnoea. Fever, hypophosphataemia and hypocalcaemia were the most common events in a small phase I trial. PMID- 11398912 TI - Fulvestrant. AB - Fulvestrant is a 7alpha-alkylsulphinyl analogue of estradiol that competes with endogenous estrogen for binding to the estrogen receptor. Once bound to the receptor, fulvestrant attenuates receptor dimerisation, effecting a rapid degradation of the estrogen receptor protein and inhibition of transcription. Fulvestrant is a potent inhibitor of the growth of human breast cancer cells in vitro and in vivo. It has demonstrated pure anti-estrogenic activity in animal systems. Intramuscular fulvestrant 250 mg once a month was as effective as the oral aromatase inhibitor anastrozole 1 mg/day in 2 phase III trials in postmenopausal women with advanced breast cancer who had received prior endocrine therapy. Median time to disease progression (the primary end-point) with fulvestrant and anastrozole was 5.4 and 3.4 months (North American trial) and 5.5 and 5.1 months (European trial). The median duration of response was 19.3 and 10.5 months (North American trial) and 14.3 and 14.0 months (European trial). The most common adverse events with fulvestrant are gastrointestinal disturbances and hot flushes. Fulvestrant showed similar tolerability to anastrozole in 2 phase III trials. PMID- 11398913 TI - Telithromycin. AB - Telithromycin is the first member of a new family of the macrolide-lincosamide streptogramin-B (MLS(B)) class of antimicrobials, the ketolides. It has a good spectrum of activity against respiratory pathogens, including penicillin- and erythromycin-resistant pneumococci, as well as intracellular and atypical bacteria. Furthermore, it has a low potential to select for resistance or induce cross-resistance among other MLS(B) antimicrobials. At the recommended dosage of 800 mg orally once daily, telithromycin reaches maximal plasma concentrations of about 2 mg/L. It penetrates rapidly into bronchopulmonary, tonsillar, sinus and middle ear tissues and/or fluids and achieves high concentrations at sites of infection. It also concentrates within polymorphonuclear neutrophils. In clinical trials in patients with community-acquired pneumonia (CAP), acute exacerbations of chronic bronchitis (AECB) or pharyngitis/tonsillitis caused by group A beta haemolytic streptococci, telithromycin 800 mg once daily achieved clinical cure rates of 86 to 95%. In acute maxillary sinusitis (AMS), cure rates were 73 to 91%. A 7- to 10-day regimen of telithromycin was as effective as a 10-day course of amoxicillin 1000 mg 3 times daily, clarithromycin 500 mg twice daily or a 7- to 10-day course of trovafloxacin 200 mg once daily for treating CAP. A 5-day regimen of telithromycin was as effective as a 10-day regimen of cefuroxime axetil 500 mg twice daily or amoxicillin/clavulanic acid 500/125 mg 3 times daily in AECB. A 5-day regimen of telithromycin was as effective as a 10-day regimen of clarithromycin 250 mg twice daily or phenoxymethylpenicillin 500 mg 3 times daily in pharyngitis/tonsillitis, or a 10-day regimen of amoxicillin/clavulanic acid 500/125 mg 3 times daily in patients with AMS. Telithromycin was well tolerated across all patient populations. Adverse events associated with telithromycin were generally mild to moderate in intensity and seldom led to treatment discontinuation. The most frequent adverse events were diarrhoea (13.3%) and nausea (8.1%). Other adverse events included dizziness and vomiting. PMID- 11398914 TI - Rofecoxib: a review of its use in the management of osteoarthritis, acute pain and rheumatoid arthritis. AB - Rofecoxib is a selective cyclo-oxygenase (COX)-2 inhibitor which has little or no effect on the COX-1 isoenzyme at doses up to 1000 mg/day. Rofecoxib has greater selectivity for COX-2 than celecoxib, meloxicam, diclofenac and indomethacin. In well-controlled clinical trials, rofecoxib 12.5 to 500 mg/day has been evaluated for its efficacy in the treatment of osteoarthritis, acute pain and rheumatoid arthritis [lower dosages (5 to 125 mg/day) were generally used in the chronic pain indications]. In the treatment of patients with osteoarthritis, rofecoxib was more effective in providing symptomatic relief than placebo, paracetamol (acetaminophen) and celecoxib and was similar in efficacy to ibuprofen, diclofenac, naproxen and nabumetone. Overall, both the physician's assessment of disease status and the patient's assessment of response to therapy tended to favour rofecoxib. In patients with postsurgical dental pain, pain after spinal fusion or orthopaedic surgery, or primary dysmenorrhoea, rofecoxib provided more rapid and more sustained pain relief and reduced requirements for supplemental morphine use after surgery than placebo. Rofecoxib was more efficacious than celecoxib in patients with acute dental pain and pain after spinal fusion surgery, although celecoxib may have been used at a subtherapeutic dose. In comparison with traditional nonsteroidal anti-inflammatory drugs (NSAIDs) ibuprofen, diclofenac and naproxen sodium, rofecoxib was similar in efficacy in the treatment of acute pain. Although naproxen sodium provided more rapid pain relief than rofecoxib in patients with primary dysmenorrhoea, the reverse was true after orthopaedic surgery: rofecoxib provided more rapid pain relief and less supplemental morphine was needed. Rofecoxib was as effective as naproxen in providing symptomatic relief for over 8700 patients with rheumatoid arthritis. Compared with traditional NSAID therapy, rofecoxib had a significantly lower incidence of endoscopically confirmed gastroduodenal ulceration and, in approximately 13,000 patients with osteoarthritis and rheumatoid arthritis, a lower incidence of gastrointestinal (GI) adverse events. Rofecoxib was generally well tolerated in all indications with an overall tolerability profile similar to traditional NSAIDs. The most common adverse events in rofecoxib recipients were nausea, dizziness and headache. In conclusion, rofecoxib is at least as effective as traditional NSAID therapy in providing pain relief for both chronic and acute pain conditions. Rofecoxib provides an alternative treatment option to traditional NSAID therapy in the management of symptomatic pain relief in patients with osteoarthritis. Initial data from patients with primary dysmenorrhoea and postoperative pain are promising and further trials may confirm its place in the treatment of these indications. Rofecoxib has also shown promising results in patients with rheumatoid arthritis and is likely to become a valuable addition to current drug therapy for this patient population. Importantly, rofecoxib is associated with a lower incidence of GI adverse events than traditional NSAIDs making it a primary treatment option in patients at risk of developing GI complications or patients with chronic conditions requiring long term treatment. PMID- 11398915 TI - Perindopril: an updated review of its use in hypertension. AB - Perindopril erbumine (perindopril) is a prodrug ester of perindoprilat, an angiotensin converting enzyme (ACE) inhibitor. Perindopril 4 to 8 mg once daily significantly reduces supine systolic blood pressure (SBP) and diastolic blood pressure (DBP) from baseline values in hypertensive patients. These reductions are maintained for at least 24 hours, as evidenced by trough/peak ratios of >50%. Vascular abnormalities associated with hypertension were improved or normalised during perindopril treatment. Perindopril 4 to 8 mg once daily significantly decreased carotid-femoral aortic pulse wave velocity (PWV), improved arterial compliance, reduced left ventricular mass index and, in patients with recent cerebral ischaemia and/or stroke, preserved cerebral blood flow despite significantly reducing SBP and DBP. Further research is needed to establish the significance of promising results showing that reductions in aortic PWV were associated with reduced mortality in patients with end-stage renal failure, a third of whom received perindopril. Response rates (numbers of patients with supine DBP < or = 90 mm Hg) were significantly higher with perindopril 4 to 8 mg once daily (67 to 80%) than with captopril 25 to 50 mg twice daily (44 to 57%) in 3 randomised double-blind trials. In other clinical trials, the antihypertensive effects of perindopril were similar to those of other ACE inhibitors (including enalapril) and calcium-channel antagonists. Combination treatment with perindopril and an antihypertensive agent from another treatment class provided additional benefits, either as first-line treatment or in patients failing to respond to monotherapy. Perindopril monotherapy was also effective in the elderly and in patients with hypertension and concomitant disease. Perindopril has a similar adverse event profile to that of other ACE inhibitors; cough is the most common event reported during treatment, and is also the most common adverse event responsible for treatment withdrawal. CONCLUSIONS: Perindopril is a well tolerated ACE inhibitor that is significantly better than captopril (in terms of response rates) in the treatment of hypertension, and as effective as other ACE inhibitors. Perindopril appears to reverse some of the vascular abnormalities associated with hypertension, including arterial stiffness and left ventricular hypertrophy, although further research is needed to confirm promising results regarding its ability to decrease associated cardiovascular morbidity and mortality. Results from ongoing studies will help confirm the place of perindopril in the treatment of hypertension; currently, it is an effective and well tolerated treatment for patients with mild to moderate essential hypertension. PMID- 11398916 TI - Modification of lignin for the production of new compounded materials. AB - The cell walls of woody plants are compounded materials made by in situ polymerization of a polyphenolic matrix (lignin) into a web of fibers (cellulose), a process that is catalysed by polyphenoloxidases (laccases) or peroxidases. The first attempt to transform the basic strategy of this natural process for use in human craftsmanship was the ancient lacquer method. The sap of the lacquer tree (Rhus verniciflua) contains large amounts of a phenol (urushiol), a polysaccharide and the enzyme laccase. This oil-in-water emulsion solidifies in the presence of oxygen. The Chinese began using this phenomenon for the production of highly creative artwork more than 6,000 years ago. It was the first example of an isolated enzyme being used as a catalyst to create an artificial plastic compound. In order to apply this process to the production of products on an industrial scale, an inexpensive phenol must be used, which is transferred by an enzyme to active radicals that react with different components to form a compounded material. At present, the following approaches have been studied: (1) In situ polymerization of lignin for the production of particle boards. Adhesive cure is based on the oxidative polymerization of lignin using phenoloxidases (laccase) as radical donors. This lignin-based bio-adhesive can be applied under conventional pressing conditions. The resulting particle boards meet German performance standards. By this process, 80% of the petrochemical binders in the wood-composite industry can be replaced by materials from renewable resources. (2) Enzymatic copolymerization of lignin and alkenes. In the presence of organic hydroperoxides, laccase catalyses the reaction between lignin and olefins. Detailed studies on the reaction between lignin and acrylate monomers showed that chemo-enzymatic copolymerization offers the possibility to produce defined lignin-acrylate copolymers. The system allows control of the molecular weights of the products in a way that has not been possible with chemical catalysts. This is a novel attempt to enzymatically induce grafting of polymeric side chains onto the lignin backbone, and it enables the utilization of lignin as part of new engineering materials. (3) Enzymatic activation of the middle-lamella lignin of wood fibers for the production of wood composites. The incubation of wood fibers with a phenol oxidizing enzyme results in oxidative activation of the lignin crust on the fiber surface. When such fibers are pressed together, boards are obtained which meet the German standards for medium-density fiber boards (MDF). The fibers are bound together in a way that comes close to that by which wood fibers are bound together in naturally grown wood. This process will, for the first time, yield wood composites that are produced solely from naturally grown products without any addition of resins. PMID- 11398917 TI - Microbial metabolites as eco-friendly agrochemicals for the next millennium. AB - As a result of the increasing environmental and health-related problems caused by the synthetic agrochemicals currently used, suitable and non-hazardous innovative alternatives are being sought. Antagonism and allelopathy, both in nature and in agro-ecosystems, have attracted these researchers' attention, with the main goal of using these phenomena in the biological control of weeds. This article presents a review on the use and efficacy of microbial secondary metabolites which have potential as natural herbicides, either directly or as templates for bio-rational eco-friendly agrochemicals (allelochemicals). Their merits as alternatives to synthetic chemicals and biological control agents have been highlighted for an holistic approach in integrated pest/weed management. PMID- 11398918 TI - Enhancement of Taxol production and excretion in Taxus chinensis cell culture by fungal elicitation and medium renewal. AB - An endophytic fungus, Aspergillus niger, isolated from the inner bark of a Taxus chinensis tree, was used as an elicitor to stimulate the Taxol (paclitaxel) production in a Taxus chinensis cell suspension culture. Different elicitor doses and elicitation times were tested in a batch culture; and the highest volumetric Taxol yield was achieved when 40 mg of the fungal elicitor (carbohydrate equivalent) l(-1) was added to the culture during the late exponential-growth phase. The elicitation resulted in a more than two-fold increase in the Taxol yield and about a six-fold increase in total secretion. The Taxol yield was further improved substantially by applying medium renewal and re-elicitation to the culture. In particular, with repeated medium renewal (in a way similar to medium perfusion) and a second elicitation of the culture, the volumetric Taxol yield was increased to 67.1+/-7.5 mg l(-1), which was about seven times the amount obtained in the non-elicited batch culture. The Taxol productivity of the perfusion-like culture with repeated fungal elicitation was 1.5 mg l(-1) day(-1), which was about 40% higher than that of the elicitor-treated batch culture and three times the productivity of the non-elicited batch culture. PMID- 11398919 TI - Application of oxygen vectors to Claviceps purpurea cultivation. AB - The application of a two-phase fermentation system for the production of ergot peptide alkaloids by Claviceps purpurea is described. Perfluorocarbons (PFC) are used as oxygen vectors in Claviceps fermentation for the first time. In shake flask cultivations, the inclusion of PFC in the medium brings about a five-fold increase in the total alkaloid production and a six-fold increase in the pharmaceutically important component, ergotamine. This rise cannot be correlated with the concentration of the added PFC and it is thought that the enhancement is due to a combination of factors, including the influence of PFC. Other oxygen vectors, such as several hydrocarbons, prove to be poor oxygen carriers in our study. Cultivations with PFC in a bioreactor are reproducible, the maximum total alkaloid and ergotamine production being attained on the 11th and 9th days, respectively. The relatively lower increase in the total alkaloid production in the bioreactor as compared to the shake-flasks is attributed to the unequal oxygen availability in the reactor. Processes with PFC offer the operational advantage of a five-fold reduction in aeration rate. PMID- 11398920 TI - Kinetic analysis of growth and xanthan gum production with Xanthomonas campestris on sucrose, using sequentially consumed nitrogen sources. AB - A batch fermentation strategy using Xanthomonas campestris ATCC 13951 for xanthan gum production has been established in which all essential medium components are supplied at the onset. This has been achieved using sucrose as sole sugar feedstock. Sequential consumption of nitrogen sources (soybean hydrolysates, ammonium and nitrate salts) was observed to facilitate the further optimisation of the medium. Biomass accumulation was limited by phosphate availability. Xanthan yields of more than 60% (grams of xanthan per gram of sugar) have been obtained with constant acetyl content. However, pyruvyl substitution decreased as the growth rate declined, due to the metabolic constraints specific to phosphate depletion. High rates of carbon conversion into xanthan were observed throughout the culture and the ATP/ADP ratio was not affected by the decline in the specific growth rate. PMID- 11398921 TI - Production of beta-carotene by a mutant of Rhodotorula glutinis. AB - Wild strains of Rhodotorula glutinis and R. rubra were investigated concerning their carotenoid production, proportion of beta-carotene and cell mass yield. R. glutinis NCIM 3353 produced 2.2 mg carotenoid/l in 72 h; and the amount of beta carotene was 14% (w/w) of the total carotenoid content (17 microg/g cell dry weight). It was subjected to mutagenesis using UV radiation for strain improvement. Out of 2,051 isolates screened, the yellow coloured mutant 32 produced 120-fold more beta-carotene (2,048 microg/g cell dry weight) than the parent culture in 36 h, which was 82% (w/w) of the total carotenoid content. Mutant 32 was grown on different carbon and nitrogen sources. The best yield of beta-carotene (33+/-3 mg/l) was obtained when glucose and yeast extract were supplied as carbon and nitrogen sources, respectively. Divalent cation salts further increased the total carotenoid content (66+/-2 mg/l) with beta-carotene as the major component (55+/-2%, w/w). PMID- 11398922 TI - Evaluation of a vertical flat-plate photobioreactor for outdoor biomass production and carbon dioxide bio-fixation: effects of reactor dimensions, irradiation and cell concentration on the biomass productivity and irradiation utilization efficiency. AB - Outdoor culture of the thermophilic cyanobacterium Synechocystis aquatilis SI-2 with a vertical flat-plate photobioreactor (VFPP) was studied during the period of January to August of 1999 in the northern region of Japan (Kamaishi, Iwate, 39 degrees N, 142 degrees E). The aim of this study was to investigate the CO2 fixation ability of the VFPP device under various irradiation conditions. An average biomass productivity of over 30 g m(-2) day(-1), which corresponds to a CO2 fixation rate of 50 g m(-2) day(-1), was achieved during this period with a 192-l scale culture. The effects on biomass productivity of the light path, height of the reactor, cell concentration and irradiation were also investigated. Variation of the optimal cell concentration to achieve the highest productivity for outdoor operation is discussed. A cell concentration of 1-2 g l(-1) was found to be most suitable for the irradiation range of 1-12 MJ m(-2) day(-1) under the experimental conditions used. PMID- 11398923 TI - Evaluation of plastic-composite supports in repeated fed-batch biofilm lactic acid fermentation by Lactobacillus casei. AB - A customized stirred-tank biofilm reactor was designed for plastic-composite supports (PCS). In repeated-batch studies, the PCS-biofilm reactors outperformed the suspended-cell reactors by demonstrating higher lactic acid productivities (2.45 g l(-1) h(-1) vs 1.75 g l(-1) h(-1)) and greater glucose consumption rates (3.27 g l(-1) h(-1) vs 2.09 g l(-1) h(-1)). In the repeated fed-batch studies, reactors were spiked periodically with concentrated glucose (75%) to maintain a concentration of approximately 80 g of glucose l(-1) in the bioreactor. In suspended-cell fermentations with 10 g of yeast extract (YE) l(-1) and zero, one, two, and three glucose spikes, the lactic acid productivities were 2.64, 1.58, 0.80, and 0.62 g l(-1) h(-1), respectively. In comparison, biofilm reactors with 7 g of YE l(-1) and zero, one, two, and three glucose spikes achieved lactic acid productivities of 4.20, 2.78, 0.66, and 0.94 g l(-1) h(-1), respectively. The use of nystatin (30 U ml(-1)) subdued the contaminating yeast population with no effect on the lactic acid productivity of the biofilm reactors, but it did affect productivity in the suspended-cell bioreactor. Overall, in repeated fed-batch fermentations, the biofilm reactors consistently outperformed the suspended-cell bioreactors, required less YE, and produced up to 146 g of lactic acid l(-1) with 7 g of YE l(-1), whereas the suspended-cell reactor produced 132 g l(-1) with 10 g of YE l(-1). PMID- 11398924 TI - Production of glucose-fructose oxidoreductase and ethanol by Zymomonas mobilis ATCC 29191 in medium containing corn steep liquor as a source of vitamins. AB - Different concentrations of corn steep liquor (CSL) were tested in the cultivation of Zymomonas mobilis. Cell growth, ethanol production, and the formation of glucose-fructose oxidoreductase (GFOR) and glucono-delta-lactonase (GL), the enzymes responsible for the bio-production of gluconic acid and sorbitol, were examined. The cell yields using 25 g CSL l(-1) and 40 g CSL l(-1) (Y(X,S) approximately 0.031 g g(-1)) were close to that obtained with 5 g yeast extract (YE) l(-1). With 5 g CSL l(-1) and 15 g CSL l(-1), the nutritional limitation led to smaller Y(X/S). Using 100 g CSL l(-1) produced an inhibitory effect on cell growth. Similar ethanol yields (92-95%) were calculated for each concentration of CSL and also for YE medium. The highest specific GFOR/GL activities (13.2-13.5 U g(-1) dry cell) were reached with 25 g CSL l(-1) and 40 g CSL l(-1), values comparable to that achieved with 5 g YE l(-1). The results confirm that CSL is an effective and cheap supplement for Z. mobilis medium, increasing the economic potential of a large-scale bio-production of sorbitol and gluconic acid by untreated Z. mobilis cells. The economic feasibility of the process is discussed. PMID- 11398925 TI - Establishment and characterization of cell-free translation/glycosylation in insect cell (Spodoptera frugiperda 21) extract prepared with high pressure treatment. AB - A coupled cell-free translation/glycosylation system, prepared from Spodoptera frugiperda insect cells, was established and optimized for protein production and glycosylation efficiency. Both translation and glycosylation were stimulated by addition of Mg2+, K+, ATP, GTP, creatine kinase and creatine phosphate, suggesting that glycoprotein productivity is largely determined by translation efficiency. However, high concentrations of creatine phosphate significantly inhibited translation. Spermidine stimulated both translation and glycosylation, but glycosylation required higher concentrations of spermidine than translation. Furthermore, extracts prepared at a nitrogen pressure of 10 kg/cm2 with the Mini Bomb cell disruption chamber had the highest glycoprotein productivity; and extracts prepared at the higher nitrogen pressure of 15 kg/cm2 retained glycosylation ability. While extracts prepared with the Potter-Elvehjem homogenizer could mediate translation, no glycosylation was achieved. This indicated that the posttranslational machinery might survive disruption by high pressure, but not by physical shearing force. This insect cell-free system was able to synthesize approximately 25 microg of glycosylated gp120/ml of reaction mixture. PMID- 11398926 TI - Function of the prosequence for in vivo folding and secretion of active Rhizopus oryzae lipase in Saccharomyces cerevisiae. AB - The role of the prosequence of Rhizopus oryzae lipase (ROL) with a preprosequence was analyzed by an expression system using Saccharomyces cerevisiae. When the mature portion of ROL (mROL) fused to the pre-alpha-factor leader sequence was expressed, secretion of active mROL was not observed. However, when mROL was synthesized together with the prosequence in trans (individually and coincidentally), secretion of active mROL was observed. The results indicate that the prosequence of ROL helped correct folding of mROL and its subsequent secretion from the yeast cells, and that physical linkage (cis) of the prosequence to the mature region was not prerequisite. From the expression of the ROL mutants with deletions at the N-terminal end of the prosequence together with mROL in trans, the residues from 20 to 37 in the prosequence were essential for the secretion, and those from 38 to 57 were essential for the formation of the active ROL and might play a role as an intramolecular chaperone. The results using the fragment of the prosequence confirmed that these residues (20-57) were significant for in vivo folding and secretion of active mROL. PMID- 11398927 TI - Expression level of heterologous proteins in Pichia pastoris is influenced by flask design. AB - The yeast Pichia pastoris is a convenient production system that enables expression of heterologous proteins in high amounts. As a fermentation method, shaking flasks are very popular because of their simplicity of handling and their low cost. We compared the expression level of the enzyme acetylcholinesterase in a transformed strain of P. pastoris grown in different flasks, presenting various designs but all with the same volume. A several-thousand-fold difference appeared in the expression levels; and the results could not be explained by differences between the flasks in the oxygenation of the medium. The data show that flask design is an important factor to consider for optimising fermentation processes. PMID- 11398928 TI - Isolation of ftsI and murE genes involved in peptidoglycan synthesis from Corynebacterium glutamicum. AB - Corynebacterium glutamicum is known to excrete large amounts of L-glutamic acid upon treatment by penicillin. However, the mechanism of L-glutamate overproduction by penicillin treatment is still unknown. A 5.3-kb HindIII fragment was isolated by directly introducing the C. glutamicum (Brevibacterium lactofermentum) ATCC 13869 gene library into the temperature-sensitive Escherichia coli murE mutant and selecting temperature resistant clones. Two open reading frames (ORFs) were found in this fragment: (1) murE, encoding UDP-N acetylmuramoyl-L-alanyl-D-glutamate:meso-diaminopimelate ligase, and (2)ftsI, encoding septum-peptidoglycan synthetase, one of the targets of penicillin (penicillin-binding protein 3). Both ORFs were involved in peptidoglycan synthesis. Proteins were synthesized from the C. glutamicum murE and ftsI genes, 55 kDa and 73 kDa respectively, in an in vitro protein synthesis system, using E. coli S30 extracts. PMID- 11398929 TI - Quantitative evaluation of the enhanced green fluorescent protein displayed on the cell surface of Saccharomyces cerevisiae by fluorometric and confocal laser scanning microscopic analyses. AB - The number of foreign protein molecules expressed on the cell surface of the budding yeast Saccharomyces cerevisiae by cell surface engineering was quantitatively evaluated using enhanced green fluorescent protein (EGFP). The emission from EGFP on the cell surface was affected by changes in pH. The amount of EGFP on the cell surface, displayed as alpha-agglutinin-fusion protein under control of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) promoter, was determined at the optimum pH of 7.0. The fluorometric analysis and the image analysis by confocal laser scanning microscopy (CLSM) showed a similar number of molecules displayed on the cell surface, demonstrating that 10(4)-10(5) molecules of alpha-agglutinin-fused molecules per cell were expressed. Furthermore, the amount of fluorescent protein expressed on cells harboring a multicopy plasmid was three to four times higher than that on cells harboring the gene integrated into the genome. PMID- 11398930 TI - Preparation of an organic solvent-tolerant strain from baker's yeast. AB - By using immobilized baker's yeast repeatedly in isooctane with occasional reactivation by cultivation, we succeeded in the preparation of an organic solvent-tolerant strain, named KK21, which could grow in the presence of isooctane. This is the first report on an organic solvent-tolerant strain from baker's yeast. Strain KK21 showed high tolerance to organic solvents and maintained a high and stable activity on continuous reduction of n-butyl 3 oxobutanoate in an isooctane-medium two-phase system. Although the morphology of strain KK21 was the same as that of baker's yeast, the saturated fatty acid occupancy (SFA occupancy), which is defined as the percentage of saturated fatty acids in the total fatty acids of phospholipids, of strain KK21 was significantly higher than that of parental baker's yeast when strain KK21 was grown in the presence of isooctane, suggesting that a decrease in fluidity of the cell membrane might play an important role in the tolerance to organic solvents. PMID- 11398931 TI - The fungicidal and phytotoxic properties of benomyl and PPM in supplemented agar media supporting transgenic arabidopsis plants for a Space Shuttle flight experiment. AB - Fungal contamination is a significant problem in the use of sucrose-enriched agar based media for plant culture, especially in closed habitats such as the Space Shuttle. While a variety of fungicides are commercially available, not all are equal in their effectiveness in inhibiting fungal contamination. In addition, fungicide effectiveness must be weighed against its phytotoxicity and in this case, its influence on transgene expression. In a series of experiments designed to optimize media composition for a recent shuttle mission, the fungicide benomyl and the biocide "Plant Preservative Mixture" (PPM) were evaluated for effectiveness in controlling three common fungal contaminants, as well as their impact on the growth and development of arabidopsis seedlings. Benomyl proved to be an effective inhibitor of all three contaminants in concentrations as low as 2 ppm (parts per million) within the agar medium, and no evidence of phytotoxicity was observed until concentrations exceeded 20 ppm. The biocide mix PPM was effective as a fungicide only at concentrations that had deleterious effects on arabidopsis seedlings. As a result of these findings, a concentration of 3 ppm benomyl was used in the media for experiment PGIM-01 which flew on shuttle Columbia mission STS-93 in July 1999. PMID- 11398932 TI - Constitutive and inducible hydroxylase activities involved in the degradation of naphthalene by Cunninghamella elegans. AB - The non-ligninolytic fungus Cunninghamella elegans was investigated for its ability to produce naphthalene hydroxylase (NAH) and naphthol hydroxylase (NOH) activities under various conditions. When the organism was cultivated on a rich growth medium, the mycelia exhibited significant constitutive NAH activity in the late exponential growth phase, but not in the early-exponential-growth-phase. On incubating the early-exponential-growth-phase mycelia with naphthalene, NAH activity was increased five-fold; however, this increase did not occur in the presence of the protein synthesis inhibitor cycloheximide. Since incubation of the late-phase mycelia with naphthalene did not lead to a higher degradation rate of naphthalene, mycelia in this physiological state have apparently lost the ability to induce synthesis of the enzyme exhibiting NAH activity. This is not due to an overall inability to perform de novo protein synthesis, since NOH activity, non-constitutive at all growth phases, could be induced by incubating late-phase mycelia with naphthalene. Whether inducible and constitutive NAH activity originate from one and the same enzyme remains to be elucidated. It is suggested that naphthalene oxidizing enzyme(s) may also oxidize pyrene, but not anthracene or benzo[a]pyrene, although the latter are degradable by C. elegans. PMID- 11398933 TI - Biodegradation of radiolabelled synthetic lignin (14C-DHP) and mechanical pulp in a compost environment. AB - Mineralization of radioactive synthetic lignin (14C-DHP) was studied in a compost environment at 35, 50 and 58 degrees C. Compost samples were successively extracted with water, dioxane and alkali, and the molecular weight distribution of some extracts was determined by gel permeation chromatography (GPC). Biodegradation of lignin-containing spruce groundwood (SGW) and pine sawdust was concurrently determined in controlled composting tests by measuring evolved CO2. The temperatures were the same as in the 14C-DHP mineralization experiment and bleached kraft paper, with a lignin content of 0.2%, was used as a reference. The mineralization of 14C-DHP was relatively high (23-24%) at 35 degrees C and 50 degrees C, although the mixed population of compost obviously lacks the most effective lignin degraders. At 58 degrees C the mineralization of 14C-DHP, as well as the biodegradation of SGW and sawdust, was very low, indicating that the lignin-degrading organisms of compost were inactivated at this temperature. SGW was poorly biodegradable (<40%) in controlled composting tests compared with kraft paper (77-86%) at all temperatures, which means that lignin inhibits the degradation of carbohydrates. During the incubation, water-soluble degradation products, mainly monomers and dimers, and the original 14C-DHP were either mineralized or bound to humic substances. A substantial fraction of 14C-DHP was incorporated into humin or other insolubles. PMID- 11398934 TI - Characterisation of the microbial 16S rDNA diversity of an aerobic phosphorus removal ecosystem and monitoring of its transition to nitrate respiration. AB - The microbial community of a conventional anaerobic-aerobic sequencing batch reactor was investigated by cloning and sequencing bacterial 16S rDNA. The 92 16S rDNA sequences analysed ranged across 50 different operational taxonomic units (OTU). The majority of these sequences were not closely related to known species. They belonged to 12 different groups, but essentially to the Cytophagales and the Proteobacteria beta, which represented 38% and 17% of the retrieved sequences respectively. No OTU numerically outnumbered the others. However, similarities were observed with previous reports on molecular characterisation of phosphorus accumulating ecosystems, suggesting an enrichment in microorganisms belonging to the Rhodocyclus group. Thereafter, the ability of this anaerobic-aerobic microbial community to accumulate phosphorus with nitrate as its energy source was investigated. The reactor was shifted from anaerobic-aerobic running conditions to anaerobic-anoxic conditions by injection of nitrate; and its microbial community was monitored by PCR-single strand conformation polymorphism (SSCP). The reactor maintained a good phosphorus accumulation and similar SSCP microbial community patterns for a period of 17 days, suggesting that the same microbial community was able to respire both oxygen and nitrate. However, this situation was unstable, since a breakdown in phosphorus accumulation occurred thereafter. PMID- 11398935 TI - Decolorization of molasses spent wash by the white-rot fungus Flavodon flavus, isolated from a marine habitat. AB - Flavodon flavus (Klotzsch) Ryvarden, a basidiomycete (NIOCC strain 312) isolated from decomposing leaves of a sea grass, decolorized pigments in molasses spent wash (MSW) by 80% after 8 days of incubation, when used at concentrations of 10% and 50%. Decolorizing activity was also present in media prepared with half strength seawater (equivalent to 15 ppt salinity). Decolorizing activity was seen in low-nitrogen medium, nutrient-rich medium and in sugarcane bagasse medium. The percentage decolorization of MSW was highest when glucose or sucrose was used as the carbon source in the low-nitrogen medium. The production of lignin-modifying enzymes, manganese-dependent peroxidase (MNP) and laccase decreased in a medium containing MSW. MNP production and MSW decolorization were inversely correlated, suggesting no role for MNP in MSW decolorization. The decolorization of MSW was not effective when F. flavus was immobilized in calcium alginate beads. Decolorization was achieved best in oxygenated cultures. Besides color, total phenolics and chemical oxygen demand were reduced by 50% in MSW treated with F. flavus, suggesting its potential in the bioremediation of effluents. PMID- 11398936 TI - Diffusion-weighted imaging in neonates. AB - Diffusion-weighted imaging (DWI) can readily be performed in the neonate, although currently studies remain a few years behind-those carried out on adults. DWI relies on the random diffusion of water molecules. As for the adult population, a pulsed gradient spin echo sequence (PGSE) with cardiac gating can be used to exploit the effect of diffusion on image contrast and to determine the apparent diffusion coefficient (D*) for tissues or fluids. Anisotropic properties caused by the restriction of the movement of water molecules may be demonstrated. In the neonatal brain restricted motion can be detected in both myelinated and unmyelinated white matter tracts. DWI has been used to study changes in global and focal ischaemic injury to the neonatal brain. A decreased D* may be documented after an ischaemic insult followed by a gradual increase. These changes are consistent with animal data but show a slower time course. Intervention following perinatal ischaemic injury must be started within hours. DWI detects early ischaemic injury and may therefore be a useful tool for identifying those infants who could benefit from intervention. PMID- 11398937 TI - How to perform diffusion-weighted imaging. AB - Diffusion-weighted imaging (DWI) has become an invaluable tool in the management of patients with stroke. DWI relies on detecting the random diffusion of water molecules. In normal tissues this movement may be restricted by the presence of cellular structures, which provide a barrier to free movement. This occurs in myelinated white matter, where movement is restricted more across than along fibres. This directional dependence is termed anisotropic restricted diffusion. The diffusion of water molecules can be made the dominant contrast mechanism within an image by applying large magnetic field gradients. The pulsed gradient spin echo (PGSE) sequence provides sensitivity to diffusion with gradient pulses either side of the 180 degrees refocusing pulse. This sequence is generally heavily T2 weighted. In order to detect normal anisotropic properties within the different components of the medullary core, association, commissural and projection fibres, DWI must be performed with sensitisation in at least three directions. PGSE sequences have been used to obtain the diffusion coefficient (D*), a measure of mobility at the molecular level within tissue. In acute infarction D* is increased; in brain death it is decreased. Diffusion contrast needs to be optimised in relation to the highly T2-dependent nature of PGSE sequences. This also requires a more detailed knowledge of how D* changes in disease, but information on nonischaemic neurological conditions is still very limited. PMID- 11398938 TI - Acquired inflammatory white matter disease. AB - OBJECTS: During the last decades there have been various attempts to define the characteristics of white matter inflammatory diseases. Most of the authors concerned agree in considering Schilder's myelinoclastic diffuse sclerosis (SD), Devic's neuromyelitis optica (NMO) and Balo's concentric sclerosis to be variants of typical multiple sclerosis (MS). Moreover, in childhood even typical MS presents more aggressive behaviour, with frequent evidence of giant, tumefactive, necrotic or haemorrhagic plaques. METHODS: The authors review the literature and cite atypical cases that appear to be intermediate between MS and SD, NMO and SD. CONCLUSIONS: Finally, some critical revisions are proposed concerning cases with clinical presentation, history, laboratory and MR findings intermediate between those of acute disseminated encephalomyelitis and of MS. PMID- 11398939 TI - Cerebellar imaging--an important signpost in paediatric neurology. AB - Cerebellar function has traditionally been viewed as limited to the control of voluntary movement. There is increasing clinical and experimental evidence that the cerebellum is involved in nonmotor behaviours and cognitive operations. In acquired and congenital cerebellar lesions such deficits can be demonstrated by appropriate testing. These nonmotor functions explain, at least in part, why many children with congenital cerebellar deficits or malformations first come to medical attention because of developmental delay. While the outcome of defined cerebellar syndromes, such as Dandy-Walker and Joubert, is documented, the relevance of cerebellar involvement related to prenatal (ischaemic) insults and of those observed in the context of more widespread CNS disorganization is not well understood. The cerebellum is vulnerable to prenatal infections and to many toxins and metabolic disorders, but not to perinatal and postnatal hypoxic ischaemic injury. Consideration of cerebellar involvement and analysis of imaging patterns are crucial in recognizing many neuropaediatric disorders and helpful in guiding appropriate additional investigations. PMID- 11398940 TI - Schizencephaly: correlation between the lobar topography of the cleft(s) and absence of the septum pellucidum. AB - The bipolar defects observed in schizencephalies-clefts in the hemispheric mantle on the one hand, absent septum pellucidum on the other--without any anatomic or functional continuity suggest that there is some sort of common specific vulnerability of both structures. A study of the correlation between lobar location of the clefts and involvement of the septum pellucidum was undertaken, considering the hypothesis that the septum pellucidum is the portion of a "medial medullary velum" that corresponds to the frontal lobe, while the psalterium would correspond to the parieto-occipital lobe and the fimbria corresponds to the temporal lobe. This retrospective study of 16 cases of schizencephaly properly investigated by MR discloses a perfect correlation, all cases with absent septum pellucidum having clefts into the frontal lobe, all cases with present septum pellucidum having clefts in the parietal, temporal, and occipital lobes, and only the few instances (3 cases) of overlapping findings being characterized by clefts in the central area, where the distinction between posterior frontal and anterior parietal lobes is uncertain because of the cortical dysplasia related to the clefts. Partial defects of the septum also proved to correlate closely, topographically, with the location of the clefts. Therefore, the facts confirm a segmental organization of the mantle and septal defects, suggesting a developmental rather than a destructive mechanism, which could at least be related to a segmental pattern of selective vulnerability. PMID- 11398941 TI - Functional imaging in the work-up of childhood epilepsy. AB - In children with medically intractable lesional epilepsy, surgery is deemed successful if the epileptogenic focus can be removed while major neurological functions are spared. Current techniques rely on invasive intracranial recordings. The new developments in functional imaging offer the possibility of localizing the epileptogenic focus noninvasively (PET/SPECT) and mapping cognitive functions (fMRI). Ictal SPECT shows hyperperfusion in the focus and has proved to have better localizing value than interictal PET or SPECT, which show focal hypometabolism or hypoperfusion. Ictal SPECT is useful for deciding on the placement of intracranial electrodes in extratemporal epilepsies, particularly in young children. Functional MRI has proved highly accurate for localizing motor and language networks, thus offering the possibilities of replacing the Wada test (language hemispheric lateralization) and studying postlesional brain plasticity. Despite the difficulties of functional imaging in children owing to the limited cooperation that can be expected, ethical constraints, and poor normative data, SPECT/PET and fMRI provide clinically useful information for presurgical work-up of childhood epilepsies. PMID- 11398942 TI - Hypothalamic hamartoma: the role of surgery in a series of eight patients. AB - Hypothalamic hamartoma are rare lesions. We report a new series of eight patients treated for precocious puberty (six cases) or gelastic seizures (two cases). Surgical resection was total in four cases (three pediculated and one sessile). Precocious puberty was controlled by surgical treatment in all cases. Gelastic seizures were controlled by medical treatment, but the patients did not become seizure free. We observed no mortality and no endocrinological or visual morbidity. The fact that a vascular "rete mirabilis" was observed on the surface of the lesion in our surgical material is an argument favoring a vascular mechanism in precocious puberty. Coagulation of this vascular structure can help control precocious puberty. Our series confirms that the hypothalamic hamartoma can be surgically treated when patients fail to respond to medical treatment, when the length of the treatment cannot be tolerated by the chidren and their families, and when there are uncontrolled gelastic seizures PMID- 11398943 TI - Ventriculomegaly and pericerebral CSF collection in the fetus: early stage of benign external hydrocephalus? AB - BACKGROUND AND PURPOSE: Mild ventricular dilatation or an asymmetric aspect of the atrium with prominent subrachnoid spaces is a challenging clinical condition in utero that requires prenatal MRI to rule out a destructive lesion or brain malformation. We report five cases that demonstrated benign external hydrocephalus postnatally, together with the prenatal MRI to define prenatal criteria of so-called benign external hydrocephalus. METHODS AND RESULTS: The prenatal MR images of five cases showing typical features of external hydrocephalus postnatally were reviewed. All cases showed in utero mild ventricular dilatation at the level of the atrium with enlargement of the subarachnoid spaces homo-, contra- or bilaterally, and predominantly in the parietooccipital areas. The head circumference was normal in all cases with no family history of macrocephaly. MRI did not demonstrate either abnormal signal within brain parenchyma or loss of the normal layering of the developing brain. CONCLUSION: Posterior mild ventricular dilatation and prominent subarachnoid spaces in a posterior distribution can be considered an early stage of benign external hydrocephalus that is nicely illustrated by MRI. PMID- 11398944 TI - Clinical aspects and prognosis of ependymoma in infants and children. A single institution experience. AB - Thirty-two patients (22 boys and 10 girls) with a histologically confirmed diagnosis of ependymoma were treated between 1972 and 1999. A total macroscopic resection was achieved in 16 of these patients, whereas 15 resections were classified by the surgeon as subtotal. In 1 patient a ventriculostomy was created as part of a palliative strategy. All children over 3 years old were treated with postoperative radiotherapy. Chemotherapy consisted of procarbazine, ifosfamide, etoposide, methotrexate, cisplatin and cytosine arabinoside. There was 1 perioperative death. Twenty children developed a relapse of disease within 2 months to 13 years and 1 month after the initial therapy. A maximal number of five recurrences were seen in 1 patient. The value of adjuvant chemotherapy on the prognosis of children with ependymoma seems to be limited. With regard to the poor outcome, the advisability of further treatment after multiple recurrences is debatable. PMID- 11398945 TI - Choroid plexus tumors in children and young adults: report of 16 consecutive cases. AB - Choroid plexus tumors are rare intraventricular tumors, and they represent 2-4% of brain tumors in children. This single-institution retrospective study involves 16 consecutive choroid plexus tumors: 13 papillomas and 3 carcinomas. Tumor locations were the lateral ventricles in 13 cases, the third ventricle in 2 cases and the fourth ventricle in 1 case. The mean age at presentation was 3.1 years. Two patients died of perioperative blood loss. Five-year survival was 85% with papillomas and 33% with carcinomas. None of the papillomas recurred after total tumor resection, and the functional outcome in long-term survivors after papilloma surgery was excellent in 92% of the cases. Two of the carcinoma patients had disseminated disease. Fifty percent of the patients had persistent hydrocephalus after tumor resection, and these required cerebrospinal fluid diversion. PMID- 11398946 TI - International study of emergency department care for pediatric traumatic brain injury and the role of CT scanning. AB - OBJECTS: Our objective was to investigate the use of CT and its relationship to head injury severity and age. METHOD: The multi-center group International Study of Head Injury Project (ISHIP) serves as the administrative body for research design, data collection and analysis. This is a nonrandomized prospective study of longitudinal outcomes following examination and care in emergency department in five different countries. The subjects of our study were 4,690 children from birth to 15 years of age, all of whom were systematically evaluated. Each child was medically evaluated and categorized as to injury severity, mechanism of injury and findings on X-ray and CT scan. Follow-up interview and assessment was completed for comparison with the presenting clinical state. RESULTS: CT scans were performed for 674 (14.3%) of the children: 438 scans were normal and 236 were abnormal (P<0.001). Of the children with abnormal CT scans, 23.3% had mild head injuries, 42.7% had moderate injuries, and 33.8% had severe injuries, as determined by the GCS. By age, 10.5% of the positive CTs were in children aged 0 2 years, 56.3% in 3- to 9-year-olds, and 33% in 10- to 15-year-olds; only in 2% of cases were both CT and X-ray positive. CONCLUSIONS: The majority of children did not need significant medical intervention. Physicians ordered X-ray investigations more frequently than CT scanning. The use of X-ray to decide whether or not CT is necessary is not warranted. The implications of positive CTs in mild or moderate injuries were most noteworthy, as were age-related interactions with positive CT findings. PMID- 11398947 TI - Cerebellar atrophy after severe traumatic head injury in children. AB - OBJECT: The purpose of this study was to describe late neuropathological MRI findings in pediatric severe head injury and to explore the relationship between these lesions and cognitive sequelae. METHODS: Thirteen infants with severe head trauma (Glasgow 6) were included in this investigation. Clinical examination, a battery of tests designed to assess neurophysiological status, and MRI investigations of the brain were obtained in periods ranging between 8 and 20 months after the accident. Hemosiderin deposits, encephalomalacia, and cerebellar atrophy were the most frequent traumatic sequelae. The lesions were located in frontal lobes, the basal ganglia, and the cerebellum. Six patients had cerebellar atrophy associated with frontal or temporal postraumatic lesions. Cerebellar clinical dysfunction was observed in only 3 of these patients. Performance on tests evaluating frontal lobe functions was depressed in 5 of them. CONCLUSIONS: Late MRI after severe head trauma in our pediatric population showed unexpected cerebellar atrophy. Its correlation with prefrontal dysfunction is difficult to confirm because of its association with other parenchymal post-traumatic lesions. Further research involving a larger sample of patients with brain injury of varying severity is in progress, to investigate whether cerebellar atrophy could be a consequence of severe head trauma. PMID- 11398948 TI - Perioperative management protocols for children with moyamoya disease. AB - Protocols for prevention of cerebral ischemic attacks caused by hyperventilation resulting from crying, as observed in perioperative pediatric moyamoya patients, were evaluated. The first protocol involved the use of sedation when staff were setting up the intravenous lines, performing neuroimaging studies, and controlling postoperative pain. The second involved the use of wound-handling techniques designed to ease postoperative wound care; these included steristrip closure, use of paraffin gauze and not using adhesive tapes. We compared 14 and 11 surgical cases handled before and after the protocols were introduced, respectively. The number of patients with perioperative cerebral infarction decreased from 2 to 0. Appropriate sedation reduced the incidence of transient ischemic attacks from 28.6% to 3.7%. The average postoperative hospital stay was similarly reduced, from 21.3 days to 16.1 days, as a consequence of the reduced incidence of complications. It is concluded that the perioperative risks can be minimized when invasive procedures are managed according to our protocols. PMID- 11398949 TI - Intervertebral foraminal ligaments of the lumbar spine: anatomy and biomechanics. AB - The anatomical existence of the transforaminal ligaments has been studied extensively. However, there are very few studies examining how the transforaminal ligaments could be involved in the causation of nerve root compression and the low back pain syndrome. In this article, the authors review earlier studies in an attempt to find anatomical and biomechanical correspondence between the intervertebral foraminal ligaments of the lumbar spine and the low back pain syndrome. PMID- 11398950 TI - Cerebral abscess associated with a pulmonary arteriovenous malformation in a patient suffering from alpha thalassaemia. AB - We report the case of a young girl suffering from alpha thalassaemia and presenting with a brain abscess caused by a silent pulmonary arteriovenous malformation. To our knowledge this is the first such case associated with alpha thalassaemia. Diagnostic investigations and treatment options are discussed. PMID- 11398951 TI - Metachronous neurohypophysical immature teratoma occurring 10 years after total resection of pineal mature teratoma. AB - An 18-year-old boy presented with an immature neurohypophysial teratoma occurring 10 years after total resection of a mature pineal teratoma through an occipital transtentorial approach in 1989. Thorough histological examination had revealed a mature teratoma. He developed panhypopituitarism and diabetes insipidus in 1999. Magnetic resonance imaging revealed a suprasellar tumor occupying the third ventricle. This tumor was totally resected through a frontobasal approach. Histological examination revealed an immature teratoma. This tumor occurred in a different site from the initial tumor and was considered to be de novo and thus a so-called metachronous germ cell tumor. Patients with completely resected mature teratoma require extended follow-up, including periodic magnetic resonance imaging, because of the risk of such a metachronous germ cell tumor. PMID- 11398952 TI - Acute epidural hematoma related to cesarean section in a neonate with Chiari II malformation. AB - We present a rare case of acute epidural hematoma in a newborn infant with congenital hydrocephalus that was related to Chiari II malformation. The hematoma was attributed to the application of excessive suction with a vacuum extractor during cesarean section. The clinical characteristics of neonatal epidural hematoma were analyzed after a review of 18 cases in the literature, and diagnosis and treatment are discussed with reference to the results. We propose that careful follow-up is essential in neonates with cephalohematoma, and that a computed tomography (CT) study should be performed immediately if an infant's head circumference is discovered to be enlarging or if the anterior fontanel bulges. PMID- 11398953 TI - Reversible central pontine and extrapontine myelinolysis in a 16-year-old girl. AB - A rare case of an osmotic demyelination syndrome in a 16-year-old girl is presented. MRI in the acute stage revealed a focal abnormal signal within the basis pontis and both caudate nuclei and putamina. Two years later brain lesions had disappeared on T1- and T2-weighted imaging, indicating that central pontine and extrapontine myelinolysis may be completely reversible. PMID- 11398954 TI - Cranial aneurysmal bone cyst: a diagnostic problem. With a review of the literature. AB - Aneurysmal bone cysts (ABC) are benign osteolytic lesions that are more common in young people than in adults and involve the skull only exceptionally. The origin of ABC is still debated; indeed, some authors consider ABC to be an anomalous bony reaction that is secondary to traumas or tumours. Conversely, others consider ABC to be a distinct entity. A case of a healthy young female affected by a left frontal ABC is reported here. The clinical onset was characterised by the sudden appearance of a tender and mildly painful frontal mass. Neuroradiological assessment showed a well-circumscribed lytic lesion of the frontal bone with predominantly outward extension. En bloc surgical removal of the lesion was successfully achieved; a reconstructive cranioplasty was also performed to repair the cranial defect. The rarity of the condition described, together with the absence of clear guidelines, prompted us to review the more recent literature with the twin goals of identifying radiological features and becoming able to address the diagnosis and rules for treatment of such a rare entity. PMID- 11398955 TI - A case of antenatal cerebral haemorrhage resulting from maternal alloimmunisation against fetal platelets. AB - Prenatal thrombocytopenia is a rare event and is generally due to fetal infection. In very rare cases, fetal thrombocytopenia is induced by maternal IgG directed against the fetal platelets. This alloimmunisation could lead to in utero bleeding. We now report such a case, in which fetal thrombocytopenia was complicated by a huge temporal lobe haematoma. Such a prenatal event is rare: only eight cases have been published, with only one pathologically confirmed case. Our patient is the second one in which neuropathological examination demonstrated prenatal intracerebral bleeding. PMID- 11398956 TI - Reducing postrelease risk behavior among HIV seropositive prison inmates: the health promotion program. AB - The prevalence of AIDS is five times higher among prison inmates than in the general population. Because recidivism is common and many inmates are serving short sentences for parole violation, HIV-seropositive inmates move frequently between prison and their home communities. We designed an eight-session prerelease intervention for HIV seropositive inmates to decrease sexual and drug related risk behavior and to increase use of community resources after release. The intervention sessions were delivered at the prison by community service providers. We found that a prerelease risk reduction intervention for HIV seropositive inmates was feasible. Descriptive results support the effectiveness of the program in reducing sexual and drug-related behaviors and in increasing use of community resources after release. Compared with men who signed up for the intervention but were unable to attend, men who received the intervention reported more use of community resources and less sexual and drug-related risk behavior in the months following release. We recommend dissemination and continued evaluation of this risk-reduction intervention. PMID- 11398957 TI - The Attitudes Toward Women with HIV/AIDS Scale (ATWAS): development and validation. AB - Over the past 20 years, many scales have been created to measure general attitudes toward People with HIV/AIDS (PWA). A high correlation between negative attitudes toward PWA and rejecting attitudes toward homosexuality has been demonstrated in many studies, which may suggest that although the scales posit to examine "attitudes toward PWA," they may actually tap into attitudes toward homosexuals with HIV/AIDS. These scales may be inappropriate to use for examining attitudes toward other groups of people with HIV/AIDS (e.g., heterosexual women). The present researchers developed and validated the Attitudes Toward Women with HIV/AIDS Scale (ATWAS). Principal components analysis of the ATWAS yielded a four factor structure accounting for 48.6% of the total variance of attitudes toward women with HIV/AIDS. The four factors were Child Care, Myths/Negative Stereotypes, Reproduction/Contraception issues, and Sympathy/Transmission Route. The ATWAS was found to have good internal consistency (r = .82) and construct validity. PMID- 11398958 TI - HIV risk behavior and HIV testing: a comparison of rates and associated factors among homeless and runaway adolescents in two cities. AB - This study examined differences in self-reported rates and predictors of HIV testing between homeless and runaway youths in San Diego (N = 1,102) and Los Angeles (N = 1,167). Youths aged 13-23 were recruited from agency and street sites using a stratified probability sampling design. Interviewers administered a structured survey instrument lasting 20 minutes, which assessed youths' involvement in HIV risk-related sexual and drug-use behaviors, contact with outreach workers, and other variables. Significantly more Los Angeles youths (78%) reported testing than did San Diego youths (52%; p < .001). Multivariable analyses controlling for risk behaviors, knowing someone with HIV, and contact with outreach workers indicated that the higher rates of these factors in Los Angeles did not account for the difference in testing rates between the cities. Youths in Los Angeles were still 1.85 times as likely to be tested as San Diego youths (p < .001), possibly a result of differing normative behavior and accessibility of testing services. PMID- 11398959 TI - Preferences about the characteristics of future HIV prevention products among men who have sex with men. AB - This study of men who have sex with men (MSM) examined preferences about the characteristics of a potential product for preventing sexual transmission of HIV, such as a rectal microbicide. MSM were recruited in West Hollywood, California. They self-administered a questionnaire and rated 48 product characteristics representing seven dimensions. Overall, the ratings were highest for effectiveness in preventing HIV and other sexually transmitted diseases, followed by characteristics reflecting the physical or secondary effects of the product and logistics of use. Physical attributes, convenience/accessibility, and psychological aspects had intermediate ratings; interpersonal dynamics had the lowest rating. Men with negative attitudes about using condoms to prevent HIV infection were more likely than their counterparts to prefer a product that does not reduce sexual sensation or pleasure, does not break the mood, and can be used after a sexual encounter ends. A similar pattern was observed when participants were stratified by whether or not they had engaged in unprotected anal intercourse in the past 12 months. The findings inform the development, testing, and marketing of a future HIV prevention product for MSM. PMID- 11398960 TI - "It's not what you know, but who you knew": examining the relationship between behavior change and AIDS mortality in Africa. AB - Until there is an effective vaccine, changing sexual behavior (e.g., use of condoms or fewer partners) is still the only course of action that can slow the spread of HIV for most Africans. But exactly which factors influence behavior change and how are still debated. This article examines the notion that as the HIV/AIDS epidemic strengthens and spreads through communities in Africa, and mortality mounts, behaviors that prevent transmission should be changing. We focus on men in three countries--Uganda, Kenya, and Zambia--examining determinants of their behavior change, and analyze the relative importance of knowing someone who has died of AIDS as compared with other factors such as age, education level, knowledge of HIV/AIDS, economic status, and marital status. Data from three DHS surveys in Uganda (1995), Zambia (1996), and Kenya (1998) are fitted to a model predicting behavior change. Results from this cross-sectional, multinational study suggest that married and working men aged 20-40 are significantly more likely to have changed their behavior. Personal experience of AIDS is a significant predictor of behavior change in Uganda and Zambia, and is marginally significant in Kenya. One implication in the context of the epidemic is that behavior change is partly determined by the high level of mortality experienced by African communities. A second implication is that higher levels of disclosure, or lower levels of denial of AIDS as a cause of death, may help individuals change their behavior. PMID- 11398961 TI - HIV risk behavior and risk-related characteristics of young Russian men who exchange sex for money or valuables from other men. AB - One of the world's newest HIV epidemics is emerging now in Russia and other countries of Central and Eastern Europe. We report on the HIV risk characteristics of young Russian men who exchange sex for money or valuables, a group that constitutes almost one-fourth of men surveyed recently in gay identified venues in St. Petersburg. Among 96 MSM who have sex for economic gain, most reported multiple male and female partners, 45% had unprotected anal intercourse with their male partners in the past three months, and many not only received but also gave money or valuables themselves to their male partners. Relative to men who did not give sex for economic gain (n = 326), those who did were younger (n = .0001), less well-educated (p = .0001), and more often unemployed (p = .02). They also were less knowledgeable concerning even basic HIV risk reduction steps (p = .02) and held many misconceptions about safer sex. Men who exchanged sex for economic gain had more male (p = .001) and female partners (p = .01) in the past three months than men who did not, and one-third had been treated for STDs. In the context of Russia's rapid cultural and social changes, economic turmoil, and gay communities not yet experienced in AIDS, HIV prevention programs must be tailored to risk patterns and dynamics different than those found in the gay communities of many western countries. PMID- 11398962 TI - Viral escape from the neutralizing antibody response: the lymphocytic choriomeningitis virus model. AB - In addition to CD8+ cytotoxic T lymphocyte (CTL) responses, neutralizing antibodies contribute substantially to the long-term immune control of noncytopathic viruses, as demonstrated during infection with the lymphocytic choriomeningitis virus (LCMV). The high virus load during the initial phase of an infection and the ability of this RNA virus to spontaneously acquire mutations are important prerequisites for escaping an ongoing immune response. In this context, LCMV escape from the humoral response by single point mutations in neutralizing envelope protein determinants may occur, particularly during conditions of CTL deficiency, leading to virus persistence. PMID- 11398963 TI - Recombination and gene conversion-like events may contribute to ABO gene diversity causing various phenotypes. AB - We identified five different alleles, tentatively named ABO*O301, *0302, *R102, *R103, and *A110, in Japanese individuals possessing the blood group O phenotype. These alleles lack the guanine deletion at nucleotide position 261 which is shared by a majority of O alleles. Nucleotide sequence analysis revealed that *0301 and *0302 had single nonsynonymous substitutions compared with *A101 or *A102 responsible for the A1 phenotype. Analysis of intron 6 at the ABO gene by polymerase chain reaction-single-strand conformation polymorphism and direct sequencing revealed that *R102 and *R103 had chimeric sequences of A-02 and B-02, respectively, from exons 6 to 7. In the analysis of five other chimeric alleles detected in the same manner, we identified a total of four different recombination-breakpoints within or near intron 6. When 510 unrelated Japanese were examined, the frequency of the chimeric alleles generated by recombination in intron 6 or exon 7 was estimated to be 1.7%. In addition, we found that *O301, *A110, *C101, *A111, and 35% of *A102 had a unique A-B-A chimeric sequence at intron 6, presumed to originate from a gene conversion-like event. We had previously established that *A110 also had an A-O2-A chimeric sequence around nucleotide position 646 in exon 7. Thus this allele has an A-B-A-O2-A chimeric sequence from intron 6 to exon 7 probably generated by two different gene conversions. Similar patchwork sequences around nucleotide position 646 in exon 7 were observed in two other new alleles responsible for the Ax and B3 phenotypes. Thus, the site is presumably a hotspot for gene conversion. These results indicate that both recombination and gene conversion-like events play important roles in generating ABO gene diversity. PMID- 11398964 TI - Genomic analysis of common chimpanzee major histocompatibility complex class I genes. AB - To investigate how MHC class I genes have changed in the approximately 5 million years since chimpanzees and humans diverged, we characterized six genomic fragments ranging in size from 5.1 to 6.1 kb, each containing the complete coding region, introns, and flanking regions of one of the following chimpanzee class I genes: Patr-A, Patr-E, Patr-F, Patr-G, Patr-H, and Patr-J. In humans, these genes are closely linked within the class I region and are representatives of three distinct functional categories of class I genes: the highly polymorphic Ia genes (HLA-A), the conserved Ib genes (HLA-E, HLA-F, and HLA-G), and the class I pseudogenes (HLA-H and HLA-J). Southern blot analysis of chimpanzee and human class I genes produced nearly identical patterns, suggesting that the organization and linkage of these genes differs little in the two species. Comparison of the chimpanzee fragment sequences with their human orthologues revealed structural conservation of these genes yet differences in their degree of functional constraint. This is apparent in the location and nature of the amino acid changes between species and the substantial differences in levels of divergence at functional and nonfunctional sites. Additionally, there is no correlation between patterns of divergence at these sites and intraspecific variation, an observation explained by either appreciable gene conversion or high levels of recombination, the latter unlikely given the observed strong linkage disequilibrium of these loci. PMID- 11398965 TI - Cloning of Clr, a new family of lectin-like genes localized between mouse Nkrp1a and Cd69. AB - We report the identification of a novel family of genes, named Clr, encoding C type lectin-like molecules, which maps in the natural killer (NK) gene complex (NKC) on mouse Chromosome 6. Genomic sequence analysis indicates the presence of at least seven members between Nkrpla and Cd69. By RT-PCR, at least three members of the family are expressed on interleukin-2-activated NK cells. Sequence analysis revealed complete open reading frames of 203-205 amino acids, with a carboxyl-terminal C-type lectin-like carbohydrate recognition domain (CRD). The CRDs of the Clr proteins exhibit a significant degree of homology with the known NKC-encoded NK-cell receptors. However, a key cysteine usually present in the CRD is missing in the Clr proteins, suggesting that their ligands and functions are distinct from other molecules encoded in the NKC. PMID- 11398966 TI - Comparative analysis of the promoter regions and transcriptional start sites of mouse Ly49 genes. AB - Despite numerous studies on the function of Ly49 natural killer cell receptors in the mouse, relatively little is known about how these genes are regulated at the transcriptional level. In the present study, we sequenced and compared 800 bp of the promoter region of nine Ly49 genes from C57B1/6 mice. This comparison showed that there is a high degree of sequence identity between the genes, and also revealed a region which is conserved between the mouse genes and the human Ly49L gene, indicating a potential core promoter region. This analysis also found that Ly49B and H differ from the other genes in having long interspersed repetitive sequence in their promoter region which suggests a gene conversion or rearrangement involving these two genes. In addition, we performed 5' rapid amplification of cDNA ends on four Ly49 genes to localize transcriptional start sites. These experiments showed that the transcriptional initiation sites are heterogeneous for all of the genes examined, and that a large majority of Ly49G transcripts originate from the second exon as well as its first intron. Although potential TATA boxes have been previously identified for some of the genes, we did not find evidence that a majority of transcripts initiate at the expected distance downstream of these boxes. Our data suggest that differences in the location of transcriptional start sites contribute to the observed complexity in receptor repertoire patterns. PMID- 11398967 TI - Characterization of the gene encoding mouse mast cell protease 8 (mMCP-8), and a comparative analysis of hematopoietic serine protease genes. AB - Serine proteases are important granule constituents in several of the major hematopoietic cell lineages. We present here the nucleotide sequence of the gene encoding mouse mast cell protease 8 (mMCP-8). mMCP-8 was initially isolated as a cDNA from a mouse mast cell line, but has recently been found to be expressed primarily by mouse basophils. mMCP-8 and its rat homologues, rMCP-8, -9, and -10, form a new group of mast cell/basophil proteases, which are more closely related to the T-cell granzymes and neutrophil cathepsin G than to the mast cell tryptases and chymases. A dot matrix comparison of the mMCP-8 gene with other closely related hematopoietic serine protease genes shows detectable homology only in the exonic regions of the genes. No indication for conservation in the promoter region or introns was observed. This latter finding indicates that the upstream regulatory region has evolved at a relatively high rate. However, despite the low degree of direct sequence conservation, no major differences in the sizes of introns or exons were observed between mMCP-8 and genes for the closest related hematopoietic serine proteases, the mouse T-cell granzymes and cathepsin G, indicating that after evolutionary separation from the T-cell granzymes and cathepsin G, the majority of mutations primarily involved single base pair substitutions or short insertions or deletions. PMID- 11398968 TI - The CD4 T cell-deficient mouse mutation nackt (nkt) involves a deletion in the cathepsin L (CtsI) gene. AB - We recently reported a novel autosomal recessive mouse mutation designated nackt (nkt). Homozygous mutant mice have diffuse alopecia and a marked reduction in the proportion of CD4+ T cells in the thymus and peripheral lymphoid tissues. Here we show that the CD4 T-cell deficiency is due to a defect in the thymic microenvironment rather than the hematopoietic compartment. Furthermore, we identified the molecular basis of the mutant phenotype by demonstrating that the nkt mutation represents a 118-bp deletion of the cathepsin L (Ctsl) gene which is required for degradation of the invariant chain, a critical chaperone for major histocompatibility complex class II molecules. This finding explains the similarities in skin and immune defects observed in nkt/nkt and Ctsl -/- mice. The data reported here provide further in vivo evidence that the lysosomal cysteine protease cathepsin L plays a critical role in CD4+ T-cell selection in the thymus. PMID- 11398969 TI - Molecular cloning and characterization of pig immunoreceptor DAP10 and NKG2D. AB - Pig immunoreceptor DAP10 cDNA was cloned from a peripheral blood lymphocyte (PBL) cDNA library using human DAP10 cDNA as a probe. The length of the pig DAP10 cDNA is 465 bp and it contains an open reading frame of 237 bp. The predicted polypeptide sequence is 79 amino acids, consisting of an 18-amino acid leader, a 16-amino acid extracellular domain, a 24-amino acid transmembrane segment, and a 21-amino acid cytoplasmic domain. The amino acid sequence of pig DAP10 has 68% and 78% sequence identity with human DAP10 and mouse DAP10, respectively. Pig DAP10 has a conserved aspartic acid in the transmembrane domain, two cysteines in the extracellular domain, and a phophatidylinositol-3 kinase-binding site (YxxM) in the cytoplasmic region. Genomic organization reveals that pig DAP10 comprises four exons and three introns. Pig DAP10 and DAP12 are genetically linked on Chromosome (Chr) 6 at 6q21 in opposite transcriptional orientation, separated by 152 bp. In Northern blot analysis, DAP10 transcripts were detected predominantly in lymphohematopoietic tissues. Pig NKG2D cDNA has an open reading frame of 642 bp. Its expected polypeptide sequence is 214 amino acids. Pig NKG2D has 66% sequence identity with human NKG2D and 56% identity with mouse NKG2D. The NKG2D gene maps to pig Chr 5q25. RT-PCR analysis reveals that pig NKG2D transcripts are expressed in PBLs, NK cells, macrophages, and monocytes. When transiently transfected into COS-7 cells, pig NKG2D requires DAP10 for cell surface expression. PMID- 11398970 TI - Occurrence of structural specialization of the serine protease domain of complement factor B at the emergence of jawed vertebrates and adaptive immunity. PMID- 11398971 TI - The hAT family: a versatile transposon group common to plants, fungi, animals, and man. AB - Transposons are ubiquitous mobile genetic elements found in all eu- and prokaryotic cells. The first transposon identified, the maize Activator element, belongs to the hAT family. hAT transposons have been identified in most eukaryotic lineages, including plants, fungi, animals and even man. The basic structural and functional features of this transposon family and its phylogenetic roots are discussed in detail, including a phylogenetic tree deduced from the amino acid sequence of the most conserved part of the transposon-encoded transposase. Emphasis is given to the use of hAT transposons as tools for gene tagging and insect transformation as well as to their biological function, i.e. are they selfish DNA, beneficial companions, or even both? PMID- 11398972 TI - tef: a mutation that causes telomere fusion and severe genome rearrangements in Drosophila melanogaster. AB - Telomeres are the stable ends of linear chromosomes in eukaryotes. These complex protein-nucleic acid structures are essential to maintain genomic stability and the integrity of linear chromosomes. We identified a new mutation in Drosophila that causes a high frequency of end-to-end fusions of chromosomes during mitosis and meiosis. Linear chromosomal ends appear to be essential for fusions to take place. These fusions do not resolve, leading to cycles of chromosomal breakage and rejoining and severe genome rearrangements. The gene is essential for normal cell proliferation and mutant tissue shows significant apoptosis. Our analysis suggests that the function encoded by the mutant gene is required to protect the linear ends of chromosomes. PMID- 11398973 TI - A tandemly repeated DNA sequence is associated with both knob-like heterochromatin and a highly decondensed structure in the meiotic pachytene chromosomes of rice. AB - Highly repetitive tandem DNA sequence repeats are often associated with centromeric and telomeric regions of eukaryotic chromosomes. The rice tandem repeat Os48 is organized as long arrays of a 355 bp monomer and is mainly located in the telomeric regions. The chromosomal locations of the Os48 sequence were determined by fluorescence in situ hybridization (FISH) on rice pachytene chromosomes. The majority of the Os48 loci are associated with brightly 4',6 diamidino-2-phenylindole (DAPI)-stained and knob-like heterochromatin in rice pachytene chromosomes. As with other DNA sequences located in the heterochromatic regions, the cytosines of the CG and C(A/T)G sites within the Os48 repeat are heavily methylated. Surprisingly, a proportion of the FISH signals are highly decondensed and deviate significantly from the DAPI-stained periphery of the pachytene chromosomes. This highly decondensed chromatin structure has not been reported in pachytene chromosomes prepared from alcohol/acid-fixed meiotic samples in any other eukaryotic species. The condensation of the Os48 sequences is dynamic during prophase I of meiosis. The FISH signals derived from the Os48 repeat progress from a condensed configuration between leptonema and early pachynema into a decondensed structure from middle pachynema to diakinesis, and then return to a condensed form at metaphase I. PMID- 11398974 TI - Orientation of nonrandomly segregating sex chromosomes in spermatocytes of the flea beetle, Alagoasa bicolor L. AB - In males of the flea beetle, Alagoasa bicolor L., spermatocytes have two achiasmate sex chromosomes, X and Y, each of which is approximately five times larger than the ten pairs of chiasmate autosomes. At metaphase I, these univalent sex chromosomes are located on a spindle domain separated from the autosomal spindle domain by a sheath of mitochondria. A single centriole pair is located at each pole of the spindle. In prometaphase I, each sex chromosome appears to maintain an attachment to both spindle poles via kinetochore microtubules (i.e., amphitelic orientation). Before anaphase I, this orientation changes to the syntelic orientation (both sister kinetochores connected to the same pole), perhaps by the release of microtubule attachments from the more distant pole by each of the chromosomes. The syntelic orientation just prior to anaphase I leaves each sex chromosome attached to the nearest pole via kinetochore microtubules, ensuring nonrandom segregation. As the sex chromosomes reorient, the autosomes follow in a sequential manner, starting with the bivalent closest to the sex spindle domain. We report here data that shed new light on the mechanism of this exceptional meiotic chromosome behavior. PMID- 11398975 TI - Large-scale chromatin fibers of living cells display a discontinuous functional organization. AB - We investigated the chromatin organization of living cells with a combination of recently developed approaches for histone and DNA labeling. Nucleosomal DNA was labeled with a histone H2B-GFP (green fluorescent protein) fusion protein and the chromatin organization of living HeLa cells was analyzed by high resolution confocal microscopy. Within the perinuclear and perinucleolar regions chromatin was organized into large-scale fibers of 2 to 8 microm in length and 300 to 500 nm in diameter. Within the nuclear interior we observed similar large-scale fibers, but in addition focal as well as diffuse forms of organization. Comparison with standard labeling and detection procedures revealed major differences in the chromatin organization observed. Chromatin organization revealed by the distribution of histone H2B-GFP was directly compared with the functional organization of chromatin by Cy3-dUTP labeling of DNA replicating at a specific time. DNA regions replicating at a specific time display characteristic physical and functional properties. Analysis of Cy3-labeled foci revealed that they are associated with all three forms of chromatin organization (fibrillar, focal and diffuse). In particular, Cy3-labeled foci appeared as discontinuous regions of large-scale fibers. These results demonstrate that large-scale chromatin fibers have discontinuous functional characteristics. PMID- 11398976 TI - Genetic and physical analysis of a single Festuca pratensis chromosome segment substitution in Lolium perenne. AB - Molecular marker analysis and genomic in situ hybridisation (GISH) were used to examine the process of chromosome segment introgression in BC2 diploid hybrids (2n=2x=14) between Lolium perenne and Festuca pratensis. Two genotypes having what appeared to be the same, single, introgressed chromosome segment of F. pratensis in the L. perenne background were crossed with diploid L. perenne to produce a recombinant series for the introgressed region. Physical and genetic analysis of this series showed that, while recombination seemed to be possible at all points along the chromosome arm, the rate of recombination varied depending on relative position: more recombination was detected in the interstitial region as compared with the centromeric or telomeric regions. The implications of these results for the use of GISH and molecular marker analysis in the measurement of linkage drag in backcross breeding programmes is discussed. PMID- 11398977 TI - Chromosomal polymorphism is associated with nematode parasitism in a natural population of a tropical midge. AB - A natural population of a tropical midge, Chironomus ramosus (Diptera: Chironomidae), was found to be polymorphic for a paracentric inversion (IV: 18C 19D). Based on the characteristic banding pattern of the fourth chromosome in the larval salivary gland polytene nuclei, individuals were classified as either structural homozygotes or heterozygotes. Isofemale lines were obtained and subsequently standard (S/S) and inversion (I/I) homozygotes were characterised by careful progeny testing in the laboratory. While exploring various biotic and abiotic factors that might be responsible for the maintenance of inversion polymorphism, we detected nematode (Family: Mermithidae) infections among the larval population. A detailed study indicated that the inversion polymorphism in the natural population of C. ramosus was apparently being maintained as a result of the selective pressure exerted by the nematode parasite. The corresponding pattern of increase and decrease in genotype frequencies and the relative fitness values indicated a selective advantage of inversion heterozygotes (S/I) over both homozygous types (S/S and I/I). Both empirical and experimental data suggest the strong heterotic nature of adaptation in this C. ramosus population towards nematode infection. This is the first report of its kind where inversion polymorphism has been shown to be associated with nematode parasitism. PMID- 11398978 TI - Glycosyl transfer: a history of the concept's development and view of its major contributions to biochemistry. AB - Humans have fewer protein-coding genes than expected for all the inherent complexities of development. Supplementary factors include the post-translational modification of proteins by glycosylation. The latter term plus transglycosylation, glycosyltransferases, and 'to glycosylate' are used in biochemistry as though they always existed. Instead they have a history that can bring new insights in this science area to younger investigators. The present account describes five decades of findings and ideas on enzymic saccharide synthesis leading, finally to a rational theory that will surely continue to serve the biosciences well in the future. PMID- 11398979 TI - Synthesis of 6-deoxy-L-idose and L-acovenose from 1,2: 5,6-di-O-isopropylidene alpha-D-glucofuranose. AB - A practical route toward the synthesis of 6-deoxy-L-idose and L-acovenose from 1,2:5,6-di-O-isopropylidene-alpha-D-glucofuranose is described. Key steps include the stereoselective hydrogenation of 6-deoxy-1,2:3,5-di-O-isopropylidene-alpha-D xylo-hex-5-enofuranose, regioselective protection of 6-deoxy-1,2-O-isopropylidene beta-L-idofuranose at 0-5, and epimerisation of 6-deoxy-5-O-tert butyldimethylsilyl-1,2-O-isopropylidene-beta-L-idofuranose at C-3. PMID- 11398980 TI - Synthesis of inositol glycan cyclic phosphates. AB - An efficient synthesis of tri-, tetra-, and pentasaccharide cyclic phosphates 1 5, structurally related to natural inositol phosphate glycans, is reported. The title compounds were assembled by PhSeOTf-promoted glycosylation of the known glucosamine precursor, t-butyldimethylsilyl 2-azido-3,6-di-O-benzyl-2-deoxy-beta D-glucopyranoside (8) with protected 1-methylthio mono-, di-, and trimannosides 7a-c, and, after conversion into glycosyl fluorides, Cp2ZrCl2- AgOTf-promoted glycosylation of differentially protected optically pure 1D-myo-inositol 11. The syntheses were completed by installing the cyclic phosphate moieties with methylpyridinium dichlorophosphate and finally, removal of all protecting groups by dissolving-metal reduction. PMID- 11398981 TI - NMR studies of mannitol-terminating oligosaccharides derived by reductive alkaline hydrolysis from brain glycoproteins. AB - Interest in the characterisation of O-mannosyl glycan structures has been stimulated following the identification of mannitol-terminating oligosaccharides among the chains released from mammalian proteins in nervous and muscle tissues, and by the discovery of a putative human O-mannosyl transferase. Several mass spectrometry methods have been applied to structure elucidation particularly when low amounts of oligosaccharide are available for analysis. However, when sufficient amounts are available, a combination of through-bond homo- and heteronuclear, and of through-space homonuclear NMR experiments permit the complete identification of these oligosaccharide sequences. We describe here the assignment of 1H and 13C NMR chemical shifts from such experiments for four mannitol-terminating oligosaccharide alditols, GlcNAcbeta-(1-->2)Manol, Galbeta (1-->4)GlcNAcbeta-(1-->2)Manol, Galbeta-(1-->4)[Fucalpha-(1-->3)]GlcNAcbeta-(1- >2)Manol and NeuAcalpha-(2-->3)Galbeta-(1-->4)GlcNAcbeta-(1-->2)Manol, that were released from brain glycopeptides by alkaline borohydride treatment. PMID- 11398982 TI - Novel oligosaccharides synthesized from sucrose donor and cellobiose acceptor by alternansucrase. AB - Cellobiose was tested as acceptor in the reaction catalyzed by alternansucrase (EC 2.4.1.140) from Leuconostoc mesenteroides NRRL B-23192. The oligosaccharides synthesized were compared to those obtained with dextransucrase from L. mesenteroides NRRL B-512F. With alternansucrase and dextransucrase, overall oligosaccharide synthesis yield reached 30 and 14%, respectively, showing that alternansucrase is more efficient than dextransucrase for cellobiose glucosylation. Interestingly, alternansucrase produced a series of oligosaccharides from cellobiose. Their structure was determined by mass spectrometry and [13C-1H] NMR spectroscopy. Two trisaccharides are first produced: alpha-D-glucopyranosyl-(1-->2)-[beta-D-glucopyranosyl-(1-->4)]-D glucopyranose (compound A) and alpha-D-glucopyranosyl-(1-->6)-beta-D glucopyranosyl-(1-->4)-D-glucopyranose (compound B). Then, compound B can in turn be glucosylated leading to the synthesis of a tetrasaccharide with an additional alpha-(1-->6) linkage at the non-reducing end (compound D). The presence of the alpha-(1-->3) linkage occurred only in the pentasaccharides (compounds C1 and C2) formed from tetrasaccharide D. Compounds B, C1, C2 and D were never described before. They were produced efficiently only by alternansucrase. Their presence emphasizes the difference existing in the acceptor reaction selectivity of the various glucansucrases. PMID- 11398983 TI - Structural characterisation of the exopolysaccharide produced by Streptococcus thermophilus EU20. AB - Streptococcus thermophilus EU20 when grown on skimmed milk secretes a high molecular-weight exopolysaccharide that is composed of glucose, galactose and rhamnose in a molar ratio of 2:3:2. Using chemical techniques and 1D and 2D-NMR spectroscopy (1H and 13C) the polysaccharide has been shown to possess a heptasaccharide repeating unit having the following structure: [chemical structure: see text]. Treatment of the polysaccharide with mild acid (0.5 M TFA, 100 degrees C for 1 h) liberates two oligosaccharides; the components correspond to the repeating unit and a hexasaccharide equivalent to the repeating unit minus the terminal alpha-L-Rhap. PMID- 11398984 TI - Synthesis of branched cyclomaltooligosaccharide carboxylic acids (cyclodextrin carboxylic acids) by microbial oxidation. AB - Novel branched cyclomaltooligosaccharide carboxylic acid (cyclodextrin carboxylic acid) derivatives were synthesized by microbial oxidation using Pseudogluconobacter saccharoketogenes to oxidize five types of branched cyclodextrins, including maltosyl beta-cyclodextrin (maltosyl-beta-CyD). For each novel cyclodextrin carboxylic acid derivative synthesized, the hydroxymethyl group of the terminal glucose residue in the branched part of the molecule was regiospecifically oxidized to a carboxyl group to give the corresponding uronic acid. In addition, the physicochemical properties of cyclomaltoheptaosyl-(6-->1) alpha-D-glucopyranosyl-(4-->1)-alpha-D-glucopyranosiduronic acid (GUG-beta-CyD) (1) and its sodium salt were studied more extensively, as these compounds are most likely to have a practical application. PMID- 11398985 TI - Facile synthesis of a comb-like mannohexaose: a trimer of the disaccharide repeating unit of the cell-wall mannans of Aphanoascus mephitatus and related species. AB - An efficient method for the preparation of a comb-like mannohexaose having alpha (1-->6) and alpha-(1-->2) linkages has been described using 6-O-acetyl-2-O benzoyl-3,4-di-O-benzyl-alpha-D-mannopyranosyl trichloroacetimidate as the key glycosyl donor in an 'inverse Schmidt' procedure. PMID- 11398986 TI - Synthesis of N4-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-L-asparagine analogues. n-Butyramide, 3-chloropropionamide, 3-aminopropionamide, and isovaleramide analogues. AB - The syntheses of four analogues of N4-(2-acetamido-2-deoxy-beta-D-glucopyranosyl) L-asparagine are described. Activated carboxylic acids were reacted with 2 acetamido-2-deoxy-beta-D-glucopyranosylamine. n-Butyric anhydride gave N-(2 acetamido-2-deoxy-beta-D-glucopyranosyl)-n-butyramide. 3-Chloropropionic anhydride was synthesized from 3-chloropropionic acid and gave N-(2-acetamido-2 deoxy-beta-D-glucopyranosyl)-3-chloropropionamide. Equilibration of the latter with ammonium bicarbonate gave N1-(2-acetamido-2-deoxy-beta-D-glucopyranosyl)-3 aminopropionamide. Succinimidyl isovalerate was synthesized and gave N-(2 acetamido-2-deoxy-beta-D-glucopyranosyl)-isovaleramide. PMID- 11398987 TI - Glycosylation of 2-thiohydantoin derivatives. Synthesis of some novel S-alkylated and S-glucosylated hydantoins. AB - 3-Aryl-5-((Z)-arylidene)-3-aryl-2-(2-methylthioethyl)-2-thiohydantoins 3a-f and 3 aryl-5-((Z)-arylidene)-2-(2',3',4',6'-tetra-O-acetyl-beta-D-glucopyranosyl)-2 thiohydantoins 7a-n were prepared from the reaction of 3-aryl-5-((Z)-arylidene)-2 thiohydantoins 2a-n with methylthioethyl chloride or 2',3',4',6'-tetra-O-acetyl alpha-D-glucopyranosyl bromide via three different routes. The compounds did not display any antiviral and antitumoral activity. PMID- 11398988 TI - A new fluorescent assay for sialyltransferase. AB - A new fluorescent assay for the sialyltransferase reaction was established. After incubation of the sialyltransferase reaction, the sialyloligosaccharide obtained was treated by acid hydrolysis, and then the NeuAc that was released was labeled with 1,2-diamino-4,5-methylenedioxibenzene. The fluorescent-labeled NeuAc could be estimated by HPLC (excitation: 373 nm; emission: 448 nm) and a Lineweaver-Burk plot could be plotted with the data from this analysis. PMID- 11398989 TI - Boat conformations synthesis, NMR spectroscopy, and molecular dynamics of methyl 4,6-O-benzylidene-3-deoxy-3-phthalimido-alpha-D-altropyranoside derivatives. AB - Addition of the elements of phthalimide to methyl 2,3-anhydro-4,6-O-benzylidene alpha-D-mannopyranoside (1) under fusion conditions has yielded methyl 4,6-O benzylidene-3-deoxy-3-phthalimido-alpha-D-altropyranoside (2). The conformation of the pyranose ring of 2 has been shown to be non-chair by 1H NMR spectroscopy, in contrast to the conformations of related derivatives having smaller substituents at C-3. Molecular dynamics simulations of 2 in explicit chloroform-d solvent have indicated four principal conformational possibilities. Of these, the 7C5/1S5 chair/skew boat form 2d has the lowest potential energy, and is largely consistent with the observed vicinal 1H-1H NMR coupling constants. PMID- 11398990 TI - Estrogenic activity of estradiol and its metabolites in the ER-CALUX assay with human T47D breast cells. AB - A number of metabolites of 17beta-estradiol were tested for their estrogenic activity using the ER-CA-LUX assay based on the increased expression of luciferase in exposed T47D breast cancer cells. E2beta and estrone showed similar potencies in the test, whereas E2alpha was 100 times less active. Incubation of cells with estrone (0.35 microM) resulted in the formation of E2beta, whereas the reverse reaction was observed for E2beta. The resulting equilibrium may explain the similar estrogenic potency of estrone in the test. The synthetic 17-hydroxy benzoate ester of E2beta was 3 times less active than the parent compound. The 17 hydroxy palmitate and oleate esters of E2beta, were respectively 25 and 200 times less active than the parent compound. The 2-hydroxy metabolites of E2beta and estrone showed a 5,000 to 10,000 fold lower activity. The 4-hydroxy metabolites were more potent than the 2-hydroxy metabolites, showing only a 20-200 times lower activity. The 2- and 4-methoxyesters of estrone were 700 times less active. It is concluded that the estrogenic potency of metabolites formed in cattle after treatment with E2beta, like estrone, E2alpha and especially the esters of E2beta, may be significant with respect to the potential risk of the use of estradiol for growth promotion in domestic animals in certain countries. PMID- 11398991 TI - In vitro Candidacidal effect of polymorphonuclear neutrophils of Behcet's patients with or without ocular involvement. AB - In this study our objective was to evaluate the in vitro candidacidal effect of polymorphonuclear neutrophils of Behcet's patients with or without ocular involvement (inactive phase). Fifteen patients with ocular involvement and 15 patients without ocular involvement were studied. Candidacidal assay was performed to assess the in vitro killing of Candida species by the polymorphonuclear neutrophils of both the study and control groups. In brief, peripheral venous polymorphonuclear neutrophils of healthy volunteers and patients with Behcet's disease were obtained by density gradient centrifugation. These cells were incubated with Candida spp. (Candida albicans, Candida tropicalis, Candida glabrata) for 3 h. Samples were seeded onto yeast extract peptone dextrose agar, incubated at 25 degrees C in a 5% humidified incubator for another 48 h, and formed colonies were counted. There was no significant difference between the candidacidal activity of polymorphonuclear neutrophils of healthy volunteers and of Behcet's patients with or without ocular involvement. Although various immunological abnormalities are reported in Behcet's disease, in vitro killing of Candida spp., which is one of the parameters for the phagocytic function of polymorphonuclear neutrophils, was not affected. PMID- 11398992 TI - Pseudoangiomatous hyperplasia of mammary stroma: a case of pure type after removal of fibroadenoma. AB - A case of pure pseudoangiomatous hyperplasia of the mammary stroma after removal of a fibroadenoma is described. The lesion, which was found in the right breast of a 40-year-old woman, was a well-circumscribed non-encapsulated, rubbery, lobulated mass measuring 40x40x35 mm. Histologically, it consisted predominantly of a proliferation of spindle cells with interanastomosing vascular-like arrangements in the interlobular or interductal stroma. Neither cytological atypia nor mitotic figures were observed. The stroma contained abundant collagen with focal hyalinization. Focally, epithelial elements showed mild ductal hyperplasia. There was no fibroadenoma. Immunohistochemically, the spindle cells were positive for vimentin, CD34, alpha-smooth muscle actin, muscle actin, CD34, calponin, and progesterone receptors. Ultrastructurally, many spindle cells had thin elongated cytoplasmic processes, which enveloped pseudovascular spaces containing a few collagenous fibrils, indicating fibroblastic cells. This lesion should be distinguished from other mammary fibrous and vascular lesions with which it may be confused. PMID- 11398993 TI - Predominant cultivable subgingival microbiota of healthy and HIV-infected ethnic Chinese. AB - Although clinically distinctive types of periodontal disease are known to be associated with HIV infection, the pathogenesis remains unclear. In the present study, the subgingival microflora of 21 HIV-infected and 11 HIV-free ethnic Chinese were studied using the direct microscopic and anaerobic culture methods. Motile curved rods were found to be three times higher in the HIV-infected group under direct microscopy. Otherwise, there were no significant differences between the diseased and healthy groups when analyzed either in relation to the morphotype distribution or Gram stain morphology or oxygen tolerance. The most common bacteria isolated were Capnocytophaga species followed by Prevotella loescheii, Streptococcus sanguinis, Lactobacillus spp. and Fusobacterium spp. Although there were 20 bacterial species that were strictly limited to the HIV infected group, and 5 limited to the healthy group, none of the species was a predominant isolate in either group (p>0.05). These findings agree with previous studies which report that subgingival bacteria of HIV-infected individuals (with or without periodontal disease) are similar to those found in healthy HIV-free individuals (with or without periodontal disease). Further work examining the subgingival microflora from patients with specific and/or severe forms of HIV associated periodontal disease is required to shed light on the pathogenesis of this complex clinical entity. PMID- 11398994 TI - Insulin-like growth factor-II in endocrine pancreatic tumours. Immunohistochemical, biochemical and in situ hybridization findings. AB - In earlier studies a high-molecular-weight (HMW) insulin-like growth factor-II (IGF-II) peptide was identified in adult human pancreas and localized to the insulin-producing B-cells. This peptide has now been investigated in neoplastic insulin cells. Forty endocrine pancreatic tumours and 17 pancreatic adenocarcinomas of ductal type were included in the study. All cases were investigated with immunohistochemical techniques using antibodies to IGF-II, insulin, pro-insulin, glucagon, somatostatin, pancreatic polypeptide, gastrin and vasoactive intestinal peptide (VIP). Frozen tissue from nine tumours and two normal pancreatic glands was extracted, gel separated, and quantified using radioimmunoassay. The tumours were also investigated by in situ hybridization. IGF-II-immunoreactive cells were found in nearly all the 18 insulin-producing tumours (16/18), in a minority of the other endocrine tumours, but not in pancreatic adenocarcinomas. All extracts from the endocrine tumours showed varying amounts of IGF-II and had different molecular-weight forms. The immunohistochemical and radioimmunoassay findings are both based on immunological binding and were further confirmed by Northern blot and in situ hybridization. These results show that IGF-II is expressed in insulin-producing tumours as well as in pancreatic tumours producing other peptides, in contrast to normal pancreatic islets where IGF-II is found exclusively in insulin-producing cells. PMID- 11398995 TI - Lymph node necrosis in systemic lupus erythematosus. A histopathological and immunohistochemical study of four cases. AB - The lymph node lesions of lupus lymphadenitis are characterized by necrosis sometimes accompanied by hematoxylin bodies, but only a few immunohistological analyses of this unique lesion have been reported. In this study we investigated the immunopathogenesis of these lesions. Lymph node specimens from four patients were analyzed immunohistochemically by applying recently developed monoclonal antibodies to immunocompetent cells. Necrosis occupied almost the entire lymph node in two cases (extensive type), whereas small foci of necrosis were found in the paracortex in the remaining two (localized type). No hematoxylin body formation was detected in any of the samples. Necrosis of the small muscular arteries, arterioles and venules was seen in the necrotic areas in all four cases. In one case of the localized type, necrotizing angitis was seen in a few arterioles and venules in the non-necrotic area. By immunohistology, amorphous depositions of immunoglobulins and C3 were demonstrated in the walls of the arterioles and venules in two cases. Our findings indicate that vasculitis due to local deposition of immune complexes in the blood vessels may play an important role in the pathogenesis of necrosis in lupus lymphadenitis. PMID- 11398996 TI - The effect of oral bacteria on Candida albicans germ-tube formation. AB - A total of eight bacterial isolates belonging to six species, and a select group of 12 oral Candida albicans isolates, were used to study the effect of bacteria on germ-tube formation. Briefly, each bacterial suspension (10(5-6) cells/ml) was mixed with a C. albicans suspension (10(7) cells/ml) and incubated at 37 degrees C for 90 min with bovine serum, and the percentage germ-tube-positive Candida cells was quantified using a haemocytometer, under light microscopy. In general, out of eight bacteria, Streptococcus sanguis SK21A, Streptococcus salivarius SK56, Escherichia coli ATCC 25922, and S. salivarius OBU3 suppressed germ-tube formation to varying degrees, with different C albicans isolates. Porphyromonas gingivalis Pg 50, Lactobacillus casei ATCC 7469 and Prevotella intermedia OBU4 elicited significant enhancement of germ-tube formation, whereas S. sanguis OBU 2 had no effect. E. coli ATCC 25922 was the only organism to show statistically significant suppression of germ-tube formation (p=0.0312). A significant increase in the germ tube production of C. albicans isolated from HIV-infected compared with HIV-free individuals was also noted. The current results tend to suggest that commensal and transient oral bacterial populations may selectively influence the differential expression of germ-tube-forming ability of C. albicans isolates. PMID- 11398997 TI - Growth rate and biofilm thickness of Streptococcus sobrinus and Streptococcus mutans on hydroxapatite. AB - Bacteria in biofilm and planktonic bacteria exhibit different properties. The objective of the present study was to compare the growth rates of Streptococcus sobrinus and Streptococcus mutans on different types of biofilm with their planktonic growth rate. Our experimental model consisted of hydroxyapatite beads coated with human saliva (sHA). Glucans or fructans were synthesized in situ on sHA by immobilized cell-free glucosyltransferase or fructosyltransferase isolated from oral bacteria. S. sobrinus or S. mutans was then adsorbed onto the glucan- or fructan-coated sHA and incubated for different time intervals. The depth of the developing biofilm was measured. Our results show that growth rates of S. sobrinus and S. mutans on both fructan- and glucan-coated sHA were similar during a 23 h period. In addition, the profile was similar to the growth profile of the same planktonic bacteria. The resemblance in growth rates between planktonic and biofilm bacteria may be attributed to the thin and non-dense biofilm formed in the initial stages of the biofilm formation. The thin biofilm coat, reaching a maximal depth of 11 microm, has only imposed limited diffusion restrictions, thus not affecting the growth of the bacteria in the biofilm. Our study shows that growth of bacteria on surfaces may resemble their growth in suspension if the bacteria are not embedded in a thick dense biofilm. PMID- 11398998 TI - Age of puberty: data from the United States of America. AB - In an attempt to determine whether the secular trend toward an earlier onset of puberty has continued over recent decades in the United States of America, published reports concerning the age of attainment of pubertal events have been reviewed. Such reports are very limited and vary in both design and inclusive ages of study subjects. Among females, two recent large cross-sectional studies indicate that fifty percent of females in the United States attain Tanner breast stage 2 at 9.5 to 9.7 years of age. This is younger than previously thought, although adequate earlier studies of girls in the United States are not available for comparison. These two studies also indicate that about 14% of girls attain Tanner stage 2 while 8 years of age; one study extends earlier reporting that about 6% exhibit onset of breast development while 7 years of age. There is no evidence that the age of menarche or the attainment of adult (Tanner 5) breast development has decreased over the past 30 years. The data also suggest an earlier onset of Tanner stage 2 pubic hair but no change in attainment of stage 5. Among males, pubic hair may be appearing at younger ages, but data are inadequate or too inconsistent to allow firm interpretation. The lack of standardization of genital criteria of pubertal onset in the male makes any conclusions regarding secular trends impossible. In summary, earlier secular trends over recent decades related to better health, improved nutrition or socio economic status, or any putative influence by endocrine disrupters cannot be verified. PMID- 11398999 TI - Genotoxic potential of xenobiotic growth promoters and their metabolites. AB - This paper reviews data reported in the literature as well as recent and unpublished studies from our laboratory on the metabolism and genotoxicity of the xenobiotic growth promoters 17beta-trenbolone, melengestrol acetate and zeranol. In our metabolic study, the oxidative in vitro metabolites generated by hepatic microsomes from rats, bovine and humans were analyzed by HPLC and GC/MS. 17beta Trenbolone gave rise to at least 13 monohydroxylated products, whereas 12 mono- and dihydroxylated metabolites were obtained with melengestrol acetate and at least 5 with zeranol. The genotoxic potential of the parent compounds was studied using the following endpoints: induction of HPRT mutations in cultured V79 cells and of lacI mutations in E. coli; induction of micronuclei in V79 cells; and formation of DNA adducts in cultured primary rat hepatocytes. Negative results were obtained in most of these assay systems. Only the micronucleus induction was marginally positive with 17beta-trenbolone and zeranol at near-cytotoxic concentrations. Commercial melengestrol acetate was found to contain an impurity causing apoptosis in V79 cells. The genotoxic potential of the numerous oxidative metabolites of the xenobiotic growth promoters remains to be studied. PMID- 11399000 TI - The birth rate of hypospadias in the Turku area in Finland. AB - Reports based on national registers of congenital malformations have suggested that the birth rate of hypospadias has increased during the last few decades. Register-based information may, however, have pitfalls because of changes in diagnostics, reporting accuracy and registration system. The aim of this study was to determine the current birth rate of hypospadias in Turku University Central Hospital (TUCH) in Finland. This was a prospective study on live-born boys born in TUCH from 1997 to 1999. In the total birth cohort (n=5,798) as well as in a special subcohort group (n=1,505) 0.3% of boys had hypospadias. Only one scrotal hypospadias was found in a boy who had a chromosomal anomaly. Other hypospadias were glandular or coronal. No increase was found in the birth rate of hypospadias when comparing our result with register-based data of boys born in Finland during the years 1970 to 1986 and surgically treated for hypospadias by the age of 8 years. No difference was found either from malformation register based data concerning the nationwide birth rate of hypospadias during the years 1993 to 1998. Due to differences in national registration systems between countries, prospective studies with equal assessment criteria are needed in order to make reliable international comparisons. PMID- 11399001 TI - Cognitive evaluation and functional outcome after stroke. AB - OBJECTIVE: To investigate the initial overall cognitive ability and its components as a predictor of functional improvement and ambulation during rehabilitation. Initial cognitive status is widely known to be a predictive factor in functional recovery in patients with stroke although some reports have found no such relationship. DESIGN: Baseline cognitive status was scored by Minimental State Examination and its subsections with such headings as "orientation," "registration," "attention and calculation," "recall," and "language" in 43 patients with postacute stroke, aged between 51 and 68 yr. Function was evaluated in terms of motor FIM and functional ambulation as categorized in "Adapted Patient Evaluation and Conference System" functional scale at the time of admission and discharge. RESULTS: Only total baseline Minimental State Examination score showed a significant correlation with discharge motor FIM improvement (r = 0.31, P = 0.04) and baseline orientation score correlated significantly with functional ambulation score improvement (r = 0.31, P = 0.03). In stepwise linear regression model, the same variables had an effect on similar outcome parameters. CONCLUSIONS: Cognitiveion evaluation should be taken as a whole to predict functional outcome in patients with postacute stroke, except for the baseline orientation score that seemed more predictive for ambulation. PMID- 11399002 TI - Complications of fluoroscopically guided caudal epidural injections. AB - OBJECTIVES: To assess the incidence of complications of fluoroscopically guided caudal epidural injections. DESIGN: A retrospective cohort design study in which chart review was performed on patients, who presented with radiculopathy and received fluoroscopically guided caudal epidural steroid injections. All injections were performed consecutively over a 12-mo period. An independent observer reviewed medical charts, which included a 24-hr post procedure telephone call by an ambulatory surgery center nurse, who asked a standardized questionnaire about complications after the injections. Physician follow-up office notes 1 to 3 wk after injection along with epidurograms were reviewed. RESULTS: The charts of 139 patients, who received 257 injections, were reviewed. Complications per injection included 12 episodes of insomnia the night of the injection (4.7%), 9 transient nonpositional headaches that resolved within 24 hr (3.5%), 8 increased back pain (3.1%), 6 facial flushing (2.3%), 2 vasovagal reactions (0.8%), 2 episodes of nausea (0.8%), and 1 increased leg pain (0.4%). No dural punctures occurred. CONCLUSIONS: No major complications occurred. The incidence of minor complications was 15.6% per injection. All reactions resolved without morbidity and no patient required hospitalization. PMID- 11399003 TI - Fluoroscopically guided therapeutic sacroiliac joint injections for sacroiliac joint syndrome. AB - OBJECTIVE: To investigate the outcomes resulting from the use of fluoroscopically guided therapeutic sacroiliac joint injections in patients with sacroiliac joint syndrome. DESIGN: A retrospective study design with independent clinical review was utilized. Thirty-one patients were included; each patient met specific physical examination criteria and failed to improve clinically after at least 4 wk of physical therapy. Each patient demonstrated a positive response to a fluoroscopically guided diagnostic sacroiliac joint injection. Therapeutic sacroiliac joint injections were administered in conjunction with physical therapy. Outcome measures included Oswestry scores, Visual Analog Scale pain scores, work status, and medication usage. RESULTS: Patients' symptom duration before diagnostic injection averaged 20.6 mo. An average of 2.1 therapeutic injections was administered. Follow-up data collection was obtained at an average of 94.4 wk. A significant reduction (P = 0.0014) in Oswestry disability score was observed at the time of follow-up. Visual Analog Scale pain scores were reduced (P < 0.0001) at the time of discharge and at follow-up. Work status was also significantly improved at the time of discharge (P = 0.0313) and at follow-up (P = 0.0010). A trend (P = 0.0645) toward less drug usage was observed. CONCLUSIONS: These initial findings suggest that fluoroscopically guided therapeutic sacroiliac joint injections are a clinically effective intervention in the treatment of patients with sacroiliac joint syndrome. Controlled, prospective studies are necessary to further clarify the role of therapeutic injections in this patient population. PMID- 11399004 TI - A home exercise program for tibial bone strengthening based on in vivo strain measurements. AB - OBJECTIVE: To compare the strain and strain rates generated during lower limb calisthenics with walking, an exercise that has been found to have only minimal effect on bone mass. Strengthening of bone, while it still has adaptive ability, can be achieved by exercise. Mechanical loading during physical activity produces strains and strain rates within the bones. It is thought that strain and strain rates higher than the usual provide the stimulus for the bones' adaptation. DESIGN: Three strain-gauged bone staples were inserted percutaneously in a 30 degrees rosette pattern in the medial aspect of the midtibial diaphysis of two volunteers. The principal compression, tension, shear strains, and strain rates were measured during various lower limb calisthenics and compared with those of jogging and walking. RESULTS: Zig-zag hopping was in the grouping of exercises with the highest principal compression, tension, and shear strains and compression strain rates, whereas walking was in the lowest or next-to-the-lowest grouping for all principal strain or strain rates. CONCLUSION: Zig-zag hopping, based on the high strain and strain rates that it produces, may be an optimal tibial bone-strengthening exercise. PMID- 11399005 TI - Psychosocial problems arising from home ventilation. AB - OBJECTIVE: To study psychosocial questions and problems of patients, who are chronically dependent on artificial ventilation, and their families. DESIGN: A total of 38 patients and family members (n = 43) were randomly selected. Several patients (n = 12) received respiratory support by nasal mask; the majority (n = 26) received respiratory support via a tracheotomy tube. Semistructured interviews were conducted. The Visual Analog Scale was also used. The extent to which the illness and chronic respiratory support were experienced as emotionally stressful, both by the patients and family members, was marked, on a scale ranging from 0 (light) to 10 (severe). RESULTS: Differences and similarities in the problems experienced were found among the patients and their families. Particularly in the case of patients receiving respiratory support by tracheostomy ventilation 24 hr/day, emotional stress on the family caregivers increased during the course of years. That the patient should urgently require bronchial suctioning at any moment made caregivers very anxious. However, several patients demonstrated increased skills in survival coping strategies. Despite diverse problems, the patients and their caregivers would elect to receive respiratory support, if the opportunity arose. CONCLUSIONS: In the case of chronically ill patients who receive continuous respiratory support via a tracheotomy tube, there is greater need for psychosocial support. Family members, who must be available 24 hr a day, need psychosocial support, professional home care, and voluntary assistance that have not been previously offered. PMID- 11399006 TI - Age- and gender-related differences in maximum shortening velocity of skeletal muscle fibers. AB - OBJECTIVE: To determine age- and gender-related differences in maximum unloaded shortening velocity (Vo) of Type I and IIA single muscle fibers. Muscle fibers must have a broad range of contractile velocities to generate the full range of power required for varied activities. DESIGN: Percutaneous needle biopsies of the vastus lateralis were obtained from 31 healthy subjects (n = 7 young men [YM], n = 7 young women [YW], n = 12 older men [OM], n = 12 older women [OW]). The slack test was used to determine Vo of individual fibers; 916 muscle fibers were chemically skinned. Fiber type was determined by myosin heavy chain isoform identification. RESULTS: Among men, Vo (fiber lengths/sec) was reduced with age in Type IIA fibers (OM vs. YM: 1.78 vs. 2.14; P < 0.05) but unchanged in Type I fibers. Among women, Vo was reduced with age in Type I fibers (OW vs. YW: 0.70 vs. 0.75; P < 0.05) but not IIA. OW had a lower Vo than did OM in both fiber types (Type I: OW = 0.70, OM = 0.77; Type IIA: OW = 1.51, OM = 1.78; P < 0.05). YW did not differ from YM. CONCLUSIONS: Both age and gender affect Vo. Age- and gender-related differences in Vo may partially explain the impairments in muscle function that occur with aging and the greater impairment in muscle function observed in OW compared with that observed in OM. PMID- 11399007 TI - An internship for college students in physical medicine and rehabilitation: effects on awareness, career choice, and disability perceptions. AB - OBJECTIVE: To show the impact that an internship program in Physical Medicine and Rehabilitation (PM&R) for college students has on their knowledge about the field, career choice, and perceptions about people with disabilities. DESIGN: Twelve students were selected to participate in the study. Students observed patient therapies and followed faculty and physiatry resident physicians. Students also participated in research studies in rehabilitation research. Group discussions regarding specific projects, research methods, career choice, and perceptions about disability were part of a didactic curriculum. Surveys about PM&R knowledge, attitudes toward people with disabilities, demographics, and course evaluations were administered. RESULTS: Results showed that the program increased knowledge about PM&R (P < 0.008). Premedical students missed significantly fewer questions (8.2 vs. 11.7; P = 0.04) on this survey than did other participants. Results also showed that this program affected their attitudes toward people with disabilities and student choice to pursue a career in health care. CONCLUSIONS: This type of internship experience provides an educational environment for college students to become acquainted with PM&R, interact positively with people with disabilities, and influence career choice in the allied health professions. PMID- 11399008 TI - Employment-seeking experiences of physiatry residents completing training during 1999. AB - Despite increasing the need for physiatrists, there is more pressure on physiatry departments and training programs to reduce the number of resident training slots. Of 321 residents completing their training in June 1999 and surveyed by questionnaire, 161 (50%) responded to the survey. The factors accounting for a successful job search were analyzed. Surveys of graduating residents can provide accurate information on job opportunities and career choices; this information can be compared with previous physiatry manpower projections. Increasing difficulty in the job search process signals that the supply of residents may exceed the demand. PMID- 11399009 TI - Using medicare databases for outcomes research in rehabilitation medicine. AB - With the advent of computerized billing for healthcare services, claims data have become useful in academic medicine. One of the largest suppliers of these types of data is Medicare, the US Government's healthcare insurance organization for the elderly and disabled. Because Medicare patients often require rehabilitation, this type of data can be useful in rehabilitation outcomes studies. Despite several significant limitations to Medicare billing data, Medicare claims data are powerful tools with which to analyze concepts in physical medicine and rehabilitation. PMID- 11399010 TI - Disease expression of Lyme borreliosis in northeastern France. AB - Since very little is known about the clinical expression of Lyme borreliosis in Western Europe, a 3-year prospective study was conducted that included all patients seen for suspected Lyme borreliosis at the Strasbourg University Hospital in northeastern France. The diagnosis of Lyme borreliosis was made on the basis of the presence of erythema migrans or on the basis of another suggestive clinical manifestation and laboratory confirmation. A total of 132 patients, 70 women and 62 men, mean age 54 years, had Lyme borreliosis according to these criteria. Within this study group, 77% of the patients were regularly exposed to tick bites and 64% could remember one. Erythema migrans, the most frequent clinical manifestation, occurred in 60% of the patients and was the only sign of Lyme borreliosis in 40%. Lymphocytoma and acrodermatitis chronica atrophicans were rare (1 and 3 patients, respectively). Nervous system involvement (mainly radiculoneuropathy), the second most common clinical manifestation, was found in 40% of the patients and was the only sign of Lyme borreliosis in 22%. Musculoskeletal involvement was present in 26% of the patients and was an isolated finding in 14%. During the study period, no patient was diagnosed with Lyme carditis. There was serological evidence of Lyme borreliosis in 75% of the cases and direct evidence of borrelial infection in 10 (7.5%). The results show that the clinical expression of Lyme borreliosis in northeastern France is similar to that in other European countries but different from that in North America. PMID- 11399011 TI - Clinical features of patients with invasive Eikenella corrodens infections and microbiological characteristics of the causative isolates. AB - Clinical features of 43 cases of invasive Eikenella corrodens infections diagnosed at National Taiwan University Hospital during a 6-year period (1993 1998) were analyzed. The clinical syndromes included head and neck infection (56%), pulmonary infection (23%), intra-abdominal infection (14%), cutaneous infection (5%), skeletal infection (2%), endocarditis (2%), and pelvic abscess (2%). Nearly two-thirds of the patients (63%) had pre-existing diseases. Malignancy (35%), especially of the head and neck, was the most common underlying illness. More than half of the patients (56%) had associated factors predisposing to invasive Eikenella corrodens infection. Polymicrobial infections occurred in 28 (65%) patients, with two-thirds of the concurrent isolates being streptococci (66%). Five cases were fatal, with four deaths directly attributable to invasive Eikenella corrodens infection. Antimicrobial susceptibility testing and molecular typing were performed on 23 preserved Eikenella corrodens isolates. Antimicrobial susceptibility testing showed that Eikenella corrodens isolates were susceptible to penicillin, amoxicillin, cefoxitin, cefotaxime, cefepime, ciprofloxacin, and imipenem. The isolates were resistant to clindamycin, metronidazole, cephalothin, and cefuroxime. None of the 23 isolates produced beta-lactamase. Random amplified polymorphic DNA patterns of the 23 isolates were different, suggesting that different clones of Eikenella corrodens caused these infections. PMID- 11399012 TI - Carriage of Neisseria meningitidis among household contacts of patients with meningococcal disease in New Zealand. AB - The aims of this study were to estimate carriage prevalence, identify factors predictive of carriage, and compare strains of Neisseria meningitidis isolated from patients with meningococcal disease and their household contacts. A total of 954 contacts of 160 patients had a nasopharyngeal swab and an interview relating to factors associated with carriage. The carriage prevalence was 20.4% for Neisseria meningitidis, 11.3% for serogroup B, and 2.6% for serogroup C. Age standardised carriage was higher in Maori (36.8%) than in Pacific Island (21.5%) or European/other (11.1%) ethnic groups. Factors associated with carriage were smoking, with personal smokers (odds ratio [OR] 2.5) and passive smokers (OR 1.6) having a higher carriage risk than those in smoke-free houses; ethnicity, with Maoris having a higher carriage risk than those of non-Maori or non-Pacific Island ethnicity (OR 2.2); gender, with males at higher risk than females (OR 1.7); and age, with 0-4-year-olds less likely and 15-24-year-olds more likely to be carriers than those over 25 years. Strong patient-contact clustering by meningococcal strain (chi-square1 = 16.7, P=0.00004) suggested an important role for the household setting in transmission. The low carriage prevalence of serogroup B Neisseria meningitidis among household contacts may reflect its low transmissibility but high virulence. No direct relationship was found between prevalence of ethnic-specific carriage and the incidence of meningococcal disease. PMID- 11399013 TI - Cluster analysis of Neisseria meningitidis type 22 strains isolated in Poland. AB - The aim of this study was to investigate the genetic relationships among strains belonging to Neisseria meningitidis serotype 22 recovered in Poland from 1995 to 1998. Phenotypical characterisation of meningococcal strains isolated from patients with meningococcal disease has been performed since 1995. Strains belonging to Neisseria meningitidis type 22 were most frequently recovered from severe cases of meningitis and septicaemia. The type 22 strains were separated into clusters by serosubtyping and DNA analyses by pulsed-field gel electrophoresis and ribotyping. However, the grouping according to serosubtypes was quite different from that obtained by ribotyping and pulsed-field gel electrophoresis. Furthermore, the two genotyping methods seldom identified the same clusters, nor were clusters identified among 26 Neisseria meningitidis type 22 strains recovered from patients hospitalised in five cities. Determination of the susceptibility to penicillin showed that a single strain was resistant (MIC, 2 mg/l) and that one-third (24/71) of the strains tested had reduced susceptibility (MIC range, 0.1-1 mg/l). PMID- 11399014 TI - Ability of three IgG-avidity assays to exclude recent cytomegalovirus infection. AB - At present, the measurement of IgG avidity appears to be the best method for differentiating primary from nonprimary cytomegalovirus (CMV) infection. This study compared the performances of three denaturation assays for the measurement of CMV IgG avidity: an in-house method and the commercially available assays Enzygnost (Dade-Behring, Germany) and Vidas (bioMerieux, France). The ability of these assays to exclude or to detect a recent CMV infection were calculated according to the results obtained in two control groups of pregnant women: 49 who had seroconverted and 80 with past infections. All three assays demonstrated a good ability to detect a recent infection (98-100%). The Dade-Behring test, in its present form, appears to be ineffective in excluding a recent CMV infection (exclusion ability: 30%), while the in-house method (exclusion ability: 96%) and the bioMerieux method (exclusion ability: 82%) performed better. The practical use of the in-house and the bioMerieux assays was evaluated in 80 women with CMV specific IgG and a positive IgM result but without documented seroconversion. At the recommended diagnostic thresholds, the concordance between these two tests was 70%. Larger studies will allow more precise determination of the capacities of both assays and specification of the diagnostic thresholds or grey areas to be used. PMID- 11399015 TI - Usefulness of route of transmission, absolute CD8+ T-cell counts, and levels of serum tumor necrosis factor alpha as predictors of survival of HIV-1-infected patients with very low CD4 + T-cell counts. AB - Potential cofactors of survival in HIV-1-infected patients with CD4+ T-cell counts of < or = 100 cells/microl were investigated. All 132 patients with CD4+ T cell counts of < or = 100 cells/microl were selected from 416 patients included in an antiretroviral therapy cohort (1989-1999). Fifty of 54 deaths were due to AIDS. There were significant associations (P<0.05) between survival and CD8+ T cell counts, clinical AIDS stage, risk group, and antiretroviral drug regimen after baseline, but only the use of protease inhibitors had an independent effect on survival (hazard ratio [HR], 0.096; 95% confidence interval [95%CI], 0.094 0.097). A substudy restricted to the cohort of 108 patients never exposed to PIs detected independent associations between survival and CD8+ T-cell counts (P=0.0016), experience with antiretroviral therapy before baseline (HR, 2.52; 95%CI, 1.31-4.82), sexual risk group for HIV infection (HR, 3.7; 95%CI, 1.92 7.12), and levels of serum tumor necrosis factor alpha (P=0.02). This study confirms that the use of PI-containing antiretroviral regimens strongly predicts survival of HIV-1-infected patients with very low CD4+ T-cell counts. When the study was restricted to patients never exposed to PIs, the parenteral route of disease transmission, high absolute CD8 + T-cell counts, and low serum levels of tumor necrosis factor alpha were independent predictors of survival in extremely advanced HIV-1 disease. PMID- 11399016 TI - Risk factors for catheter-associated bacteriuria in a medical intensive care unit. AB - In a prospective study including 137 consecutive catheterised patients in a medical intensive care unit, the following variables were analysed as possible risk factors for catheter-associated bacteriuria, defined as a quantitative culture with > or = 10(5) organisms/ml: age, sex, simplified acute and physiologic score at admission, duration of catheterisation, diabetes mellitus, immunosuppression, neurologic disorders and prior systemic antibiotic exposure during hospitalisation. The frequency of catheter-associated bacteriuria was 30.7%. By multivariate analysis, female sex (odds ratio [OR], 5.1; 95% confidence interval [CI], 1.9-13.5; P=0.001) and a duration of catheterisation >11 days (OR, 19.4; 95% CI, 5.5-68.7; P=0.0001) were risk factors for catheter-associated bacteriuria, and prior antibiotic exposure was a protective factor (OR, 0.06; 95% CI, 0.019-0.21; P=0.0001). PMID- 11399017 TI - Lemierre's syndrome with spondylitis and pulmonary and gluteal abscesses associated with Mycoplasma pneumoniae pneumonia. AB - Lemierre's syndrome, a systemic anaerobic infection caused by Fusobacterium necrophorum, is characterized by an acute oropharyngeal infection, septic thrombophlebitis of the internal jugular veins, sepsis, and multiple metastatic infections. It commonly leads to pulmonary parenchymal abscesses and occasionally to septic arthritis, osteomyelitis, or liver or spleen abscesses. Reported here is a case of spondylitis and pulmonary and gluteal abscesses that occurred as part of a classic presentation of Lemierre's syndrome. Treatment with imipenem and clindamycin was successful. PMID- 11399018 TI - Improved methods for detection of methicillin-resistant Staphylococcus aureus. AB - In order to assess the performance of two detection methods, a set of 93 recent clinical isolates of Staphylococcus aureus, including a large number of strains that demonstrated low-level methicillin-resistance were evaluated using the MRSA Screen (Denka Seiken, Japan), a commercial latex agglutination test to detect penicillin-binding protein 2' (PBP2'), and a polymerase chain reaction assay using the LightCycler Instrument (Roche Diagnostics, Switzerland). The results show that the latex agglutination test is highly sensitive if performed after induction by cefoxitin. Inconclusive results can be rapidly confirmed on the same day by real-time polymerase chain reaction used to detect mecA and femA genes. PMID- 11399019 TI - In vitro activity of gemifloxacin (SB-265805) compared to eleven other antimicrobial agents against streptococcal isolates, excluding Streptococcus pneumoniae. AB - The purpose of the study presented here was to determine the in vitro activity of gemifloxacin compared with that of 11 other antimicrobial agents (5 of them quinolones) against 400 isolates of beta-haemolytic and viridans group streptococci. The minimum inhibitory concentration values for gemifloxacin against 90% of the streptococci tested were as follows: Lancefield groups A, C and G, 0.06 microg/ml; Lancefield group B, Streptococcus mitis, Streptococcus mutans and Streptococcus bovis, 0.125 microg/ml; and Streptococcus milleri, 0.03 microg/ml. Resistance to penicillin, ampicillin and erythromycin was found mainly in the Streptococcus mitis isolates; tetracycline showed variable results, and no vancomycin resistance was encountered. Higher rates of ciprofloxacin resistance were identified in the Streptococcus bovis, mitis and mutans isolates. In conclusion, gemifloxacin was the most active quinolone tested followed by trovafloxacin, sparfloxacin, grepafloxacin, ciprofloxacin and levofloxacin, especially against isolates resistant to beta-lactam agents, macrolides and tetracycline. PMID- 11399020 TI - Flucytosine primary resistance in Candida species and Cryptococcus neoformans. AB - The in vitro activity of flucytosine (5FC) against 1,140 clinical isolates of Candida spp. and Cryptococcus neoformans was evaluated and compared with the activity of amphotericin B, fluconazole and itraconazole. Overall, 87.72% (1,000/1,140) of yeasts were susceptible to 5FC. This agent showed less potent in vitro activity against Candida glabrata, Candida krusei, Candida guilliermondii and Cryptococcus neoformans (MIC90s, 8-16 microg/ml) and intermediate activity or resistance to 6.5% of Candida albicans, 5.1% of Candida tropicalis and 0.8% of Candida parapsilosis strains. Amphotericin B showed potent activity against isolates with an MIC of 5FC > or = 8 microg/ml. A total of 112 of 140 strains that were SFC-intermediate or -resistant showed decreased susceptibility to azoles (P < 0.01). PMID- 11399021 TI - Evaluation of an optical immunoassay for the rapid detection of influenza A and B viral antigens. AB - An optical immunoassay for the rapid detection of influenza types A and B viral antigens, FLU OIA (Biostar, USA), was prospectively compared with antigen detection methods and cell culture on 400 respiratory specimens during an influenza outbreak that occurred in Switzerland in 1998/1999. The FLU OIA had an overall sensitivity of 64.4% (95%CI, 56.3-71.7%) and a specificity of 94.9% (95%CI, 89.8-97.7%). Using specimens from pediatric and adolescent patients, the sensitivity obtained (71.8%; 95%CI, 61.7-80%) was different than that achieved with specimens from adult patients (51.4%; 95%CI, 36.5-65%) (P=0.004). The results show that rapid diagnostic tests with higher sensitivity and specificity for the detection of influenza virus are needed. PMID- 11399022 TI - Patterns of Plasmodium falciparum drug resistance in nonimmune travellers to Africa. AB - Falciparum malaria is a major cause of morbidity and mortality among tourists returning from endemic areas. Treatment of infected travellers is frequently standardised, and resistance tests are rarely done as they are difficult to perform and interpret. In vitro tests of resistance to chloroquine, mefloquine, quinine, halofantrine, tetracycline, and sulfadoxine/pyrimethamine were performed on Plasmodium falciparum isolates obtained from 52 German travellers returning from malarious areas in sub-Saharan Africa. All patients were treated successfully with a standard dose of mefloquine. No in vivo resistance was observed. Although 20 (38.5%) isolates showed no signs of in vitro resistance, the remaining 32 (61.5%) were resistant to at least one of the tested antimalarial agents. Of these 32 isolates, 11 were multiresistant. Resistance to chloroquine was most frequently observed (55.8%), followed by sulfadoxine/pyrimethamine (11.5%), mefloquine (9.6%), quinine (3.8%), and halofantrine (1.9%). In vitro resistance to tetracycline was not detected in this study group. Treatment of falciparum malaria without resistance testing appears to be effective in most cases, but possible therapeutic failure due to emerging drug resistance should be kept in mind. PMID- 11399023 TI - Effect of pneumococcal vaccine on plasma HIV-1 RNA of stable patients undergoing effective highly active antiretroviral therapy. PMID- 11399024 TI - In vitro development of resistance to newer fluoroquinolones in Streptococcus pneumoniae isolates with reduced susceptibility to ciprofloxacin. PMID- 11399025 TI - Seroprevalence of human herpesvirus-8 in different patient groups in Cambodia and Germany. PMID- 11399026 TI - Evaluation of novel enzyme immunoassay for detection of Helicobacter pylori antigen in stool. PMID- 11399027 TI - Comparison of a rotavirus latex agglutination test with two rapid immunochromatographic test devices for detection of rotavirus in human feces. PMID- 11399028 TI - Identification of human liver diacetyl reductases by nano-liquid chromatography/Fourier transform ion cyclotron resonance mass spectrometry. AB - Several forms of diacetyl-reducing enzyme were found to exist in the human liver cytosol. Three (DAR-2, DAR-5, and DAR-7) of them were purified as a single band on SDS-PAGE by a combination of a few kinds of column chromatographies. The in gel tryptic digests of the purified enzymes were analyzed by nano-liquid chromatography (LC)/Fourier transform ion cyclotron resonance mass spectrometry (FT ICR MS), which provided peptide masses at a ppm-level accuracy. The enzymes, DAR-2, DAR-5, and DAR-7, were identified as alcohol dehydrogenase beta subunit (ADH2), carbonyl reductase (CBR1), and aldehyde reductase (AKR1A1), respectively, by peptide mass fingerprinting. In addition, an alternating-scan acquisition of nano-LC/FT ICR mass spectra, i.e., switching of normal acquisition conditions and in-source fragmentation conditions scan by scan, provided sets of parent and fragment ion masses of many of the tryptic peptides in a single LC/MS run. The peptide sequence-tag information at the ppm-level accuracy was used to further confirm the protein identities. It was demonstrated that nano-LC/FT ICR MS can be used for rigorous protein identification at a subpicomole level as an alternative technique to nano-LC/MS/MS. PMID- 11399029 TI - Characterization of a mouse monoclonal antibody specific for O-linked N acetylglucosamine. AB - beta-O-linked N-acetylglucosamine (O-GlcNAc) is an abundant posttranslational modification of resident nuclear and cytoplasmic proteins in eukaryotes. Increasing evidence suggests that O-GlcNAc plays a regulatory role in numerous cellular processes. Here we report on the production and characterization of a highly specific mouse monoclonal antibody, MAb CTD110.6, that specifically reacts with O-GlcNAc. The antibody recognizes O-GlcNAc in beta-O-glycosidic linkage to both serine and threonine. We could detect no cross-reactivity with alpha-linked Ser/Thr-O-GlcNAc, alpha-linked Ser-O-linked N-acetylgalactosamine (O-GalNAc), or N-linked oligosaccharides on ovalbumin and immunoglobulin G. The monosaccharide GlcNAc, but not GalNAc, abolishes immunoreactivity, further demonstrating specificity toward O-GlcNAc. Furthermore, galactose capping of O-GlcNAc sites also inhibits CTD110.6 immunoreactivity. Enrichment of GlcNAc-containing glycoproteins using the lectin wheat germ agglutinin dramatically enriches for CTD110.6-reactive proteins. The antibody reacts with a large number of proteins from cytoplasmic and nuclear extracts and readily detects in vivo changes in O GlcNAc modification. These studies demonstrate that CTD110.6 is highly specific toward O-GlcNAc, with no cross-reactivity toward similar carbohydrate antigens or toward peptide determinants. PMID- 11399030 TI - Characterization of multicomponent monosaccharide solutions using an enzyme-based sensor array. AB - We report the development of a sensor for rapidly and simultaneously measuring multiple sugars in aqueous samples. In this strategy, enzyme-based assays are localized within an array of individually addressable sites on a micromachined silicon chip. Microspheres derivatized with monosaccharide-specific dehydrogenases are distributed to pyramidal cavities anisotropically etched in a wafer of silicon (100) and are exposed to sample solution that is forced through the cavities by a liquid chromatography pumping system. Production of fluorescent reporter molecules is monitored under stopped-flow conditions when localized dehydrogenase enzyme systems are exposed to their target sugars. We demonstrate the capability of this analysis strategy to quantify beta-D-glucose and beta-D galactose at low micromolar to millimolar levels, with no detectable cross-talk between assay sites. Analysis is achieved either through fluorescence detection of an initial dehydrogenase product (NADH, NADPH) or by production of a secondary fluorescent product created by hydride transfer from the reduced nicotinamide cofactor to a fluorogenic reagent. The array format of this sensor provides capabilities for redundant analysis of sugars and for monitoring levels of other solution components known to affect the activity of enzymes. The use of this strategy to normalize raw fluorescence signals is demonstrated by the determination of glucose and pH on a single chip. Alternatively, uncertainties in the activity of an immobilized enzyme can be accounted for using standard additions, an approach used here in the determination of serum glucose. PMID- 11399031 TI - Observation of charge state and conformational change in immobilized protein using surface plasmon resonance sensor. AB - Behaviors of proteins immobilized on a solid surface were investigated using BIACORE, a biosensor utilizing surface plasmon resonance. This sensor is usually used for analyzing binding events during biomolecular interactions. Here we propose a novel use of this sensor to monitor two kinds of intramolecular changes in immobilized proteins. Several proteins were covalently attached to dextran chains on the sensor surface in the flow cell and were then exposed to a series of buffers with varying pH. Signal changes derived from changes of refractive index around the sensor surface were detected during and after the exposure to each of these buffers, which we denoted as in situ values and postvalues, respectively. The in situ value reflects the behavior of immobilized proteins in these buffers and was revealed to have a correlation with total charge state of the proteins, while the postvalue reflects how immobilized proteins react after the exposure and was suggested to represent the degree of conformational changes of the proteins. This method is expected to be applicable to various analyses and can provide us with new information about the behavior of proteins on solid phase. PMID- 11399032 TI - BIACORE data processing: an evaluation of the global fitting procedure. AB - Data from real-time molecular interaction analysis using BIACORE are currently evaluated by numerical integration. We have investigated the ability of two software packages (BIAevaluation 3.0 and CLAMP99) to analyze complex interactions. Three experimental data sets of high quality obtained with BIACORE upgraded and 2000 instruments, representative of simple bimolecular, heterogeneous ligand, and mass-transport-limited interactions, were processed by the global fitting procedure. The two software, which differ mainly in the statistical assessment of the output values, were able to discriminate correctly between various interacting models and provided very close output parameters with satisfactory statistical tests. PMID- 11399033 TI - An enzymatic assay for quantifying sphingomyelin in tissues and plasma from humans and mice with Niemann-Pick disease. AB - Sphingomyelin is an important lipid component of cell membranes and lipoproteins which can be hydrolyzed by sphingomyelinases into ceramide and phosphorylcholine. The type A and B forms of Niemann-Pick disease (NPD) are lipid storage disorders due to the deficient activity of the enzyme acid sphingomyelinase, and the resultant accumulation of sphingomyelin in cells and tissues. In this paper we report a new, enzyme-based method to quantify the levels of sphingomyelin in tissues and plasma of normal individuals and NPD patients. The method utilizes sphingomyelinase from Bacillus cereus to completely hydrolyze the sphingomyelin into ceramide. Quantification of the sphingomyelin-derived ceramide is accomplished using Escherichia coli diacylglycerol (DAG) kinase and [gamma (32)P]ATP. The resulting [(32)P]ceramide is quantified using a phosphor-imager system following TLC separation. This procedure allowed quantification of sphingomyelin over a broad range from 10 pmol to 1 nmol. To validate this assay we quantified sphingomyelin in plasma and tissues obtained from normal and NPD mice and humans. The sphingomyelin content in adult homozygous (-/-) or heterozygous (+/-) NPD mouse plasma was significantly elevated compared to that of normal mice (up to twofold). Moreover, the accumulated sphingomyelin in the tissues of NPD mice was 4 to 40 times higher than that in normal mice depending on the tissue analyzed. The sphingomyelin levels in plasma from several type B NPD patients also were significantly elevated compared to normal individuals of the same age. Based on these results we propose that this new, enzyme-based procedure can provide sensitive and reproducible sphingomyelin quantification in tissues and fluids from normal individuals and NPD patients. It could be a useful tool for the diagnosis of NPD and the evaluation of NPD treatment protocols, as well as for the study of ceramide-mediated apoptosis since the method provides the simultaneous determination of sphingomyelin and ceramide in the same lipid extract. PMID- 11399034 TI - A highly sensitive method for measurement of myosin ATPase activity by reversed phase high-performance liquid chromatography. AB - A new method for measurement of myosin ATPase activity has been developed utilizing reversed-phase high-performance liquid chromatography (HPLC), which detects as low as 0.05 nmol of ADP hydrolyzed from ATP. After termination of the ATPase reaction by addition of perchloric acid, the hydrolysate ADP and substrate ATP were separated by reversed-phase HPLC. The absorbance of ADP was monitored at 259 nm, and the amount of ADP was quantified from its peak area on the chromatogram by use of the NIH Image computer software. Our method showed linearity over a wide range from 0.05 to 10 nmol of ADP per 20 microl with a coefficient of determination (r(2)) of 0.99. Myosin ATPase activities determined by the HPLC method were almost identical to those determined by the malachite green method, a widely used spectrophotometric method with range of detection from 1 to 8 nmol of phosphate. Because our method requires only a small volume of reaction solution, it will be a powerful tool for measuring ATPase activity of motor proteins, which are difficult to obtain in large amount. PMID- 11399035 TI - Differential centrifugation separates cardiac sarcolemmal and endosomal membranes from Langendorff-perfused rat hearts. AB - The application of subcellular fractionation protocols developed in soft tissues to fibrous organs such as the heart is unsuitable given the substantial differences in subcellular structure these tissues exhibit. The purpose of this study was to develop a simple method for the separation of sarcolemma and endosomes from isolated Langendorff-perfused rat hearts. Hearts were homogenized with either an Ultra-Turrax homogenizer or a hand-held glass tissue grinder. Quantitative immunoblots assessed the enrichment of the sarcolemmal proteins caveolin 3 and the sodium potassium ATPase and the endosomal proteins rab4 and GLUT4 in different membrane fractions. Application of homogenates to sucrose and Percoll density gradients failed to resolve membranes differentially enriched in sarcolemmal or endosomal marker proteins, indicating little difference in density between the sarcolemma and endosomes. However, successive spins of homogenates from a hand-held glass tissue grinder successfully separated the endosomes from the sarcolemma, indicating differences in masses between the two membrane fractions. Approximately 70% of total caveolin 3 and sodium potassium ATPase immunoreactivity was in membrane pellets up to 20,000g and approximately 85% of rab4 and GLUT4 in pellets from 20,000-100,000g. In addition, 86% of ouabain sensitive ATPase activity (sodium potassium ATPase activity) was in membrane pellets up to 20,000g. Therefore, sarcolemmal membranes were pelleted up to 20,000g, and endosomal membranes between 20,000 and 100,000g. Regional ischemia (40 min) followed by reperfusion (60 min) caused the translocation of GLUT4 (but not rab4) from the endosomal membranes to the sarcolemma in the area of the heart subjected to ischemia. PMID- 11399036 TI - Quantitative detection in the attomole range for immunochromatographic tests by means of a flatbed scanner. AB - This work describes the use of the combination of carbon black as an antibody label, a membrane-based immunochromatographic device, and a flatbed scanner as a quantitative test system. The scanner detected 0.4-345 ng carbon black/mm(2) on a nitrocellulose membrane (0.2-170 amol carbon black/mm(2)) with an imprecision (coefficient of variation, CV) lower than 2% for the carbon black determination and a detection limit of 0.04 ng carbon black/mm(2) (0.02 amol/mm(2)). The detection ability was compared to that obtained with alkaline phosphatase (ALP) using a substrate yielding a chemiluminescent signal (0.02 amol ALP/well), beta galactosidase using a substrate yielding a fluorescent signal (0.3 amol beta galactosidase/well), and horseradish peroxidase (HRP) using a substrate yielding a colored signal (5 amol HRP/microtiter well). The carbon black immunochromatographic test for immunoglobulin E (IgE) showed a detection limit of 0.13 pM IgE (0.01 kU/L) after a testing time of 10 min. The scanner detection imprecision for the IgE determination was 0.6% CV in the range 1-10 kU IgE/L when 2.3 mm(2) was used for detection and 1% CV when 0.19 mm(2) was used. A flatbed scanner is an inexpensive instrument with multiple uses, which now also includes the sensitive evaluation of immunoassays. PMID- 11399037 TI - Oxygen radical absorbance capacity of the phenolic compounds in plant extracts fractionated by high-performance liquid chromatography. AB - The oxygen radical absorbance capacity (ORAC) assay has been used to quantify the antioxidative properties of phytonutrients in fruit and vegetable extracts. Using aqueous methanol extracts of tea and spinach as a model systems, separation of the components in the extracts by HPLC followed by semiautomatic ORAC analysis of the column fractions permitted the determination of peroxyl-radical-scavenging profiles, demonstrating the relative abilities of the individual extract components to scavenge peroxyl radicals. ORAC values for up to 80 HPLC fractions were measured, confirming the major contribution of epigallocatechin gallate in the peroxyl radical scavenging of green tea extracts. Although the flavonoids in spinach extracts provided resistance to peroxyl radicals, components that did not bind to the HPLC column and simple phenolic compounds may also be important contributors to the total ORAC activity of spinach leaf extracts. Application of these procedures to plants believed to provide certain human health benefits by reducing free radicals may allow the identification and characterization of the specific components responsible for the free-radical-scavenging activities. PMID- 11399038 TI - A colorimetric assay for high-throughput screening of inhibitors of herpes simplex virus type 1 alkaline nuclease. AB - Herpes simplex virus type 1 (HSV-1) encodes a deoxyribonuclease that is frequently referred to as alkaline nuclease (AN) because of its elevated pH optimum. Studies with recombinant viruses which contain deletions in the HSV-1 gene encoding AN have indicated that this enzyme is required for efficient virus replication and therefore represents a potential target for novel antiviral therapies. A simple colorimetric assay for deoxyribonuclease activity employing a DNA-methyl green substrate was adapted for use in a high-throughput screen to identify small molecule inhibitors of this enzyme. This screen identified 1,2 benzoisothiazolin-3-one as a specific inhibitor of AN, since it exhibited activity against AN but was completely inactive against bovine pancreatic DNaseI. Subsequent studies revealed that this compound most likely inhibited AN by forming disulfide linkages with one or more exposed cysteine residues on the surface of the enzyme and that AN was sensitive to sulfhydryl-group-modifying reagents in general. These results demonstrated the utility of this DNA-methyl green substrate-based assay in both the rapid identification and the characterization of novel small molecule inhibitors of the AN encoded by HSV-1 and other herpesviruses. PMID- 11399039 TI - Preferred binding sites for [N-MeCYs(3), N-MeCys(7)]TANDEM determined using a universal footprinting substrate. AB - We have prepared a novel footprinting substrate which contains all 136 tetranucleotide sequences and have used this to determine the preferred binding sites for the synthetic quinoxaline antibiotic [N-MeCys(3),N-MeCys(7)]TANDEM. We find that, although the ligand binds to all TpA steps, it binds best to the tetranucleotide sequence ATAT and shows only weak interaction with TTAA and GTAC. The best binding sites contain the sequences ATAX and XTAT. PMID- 11399040 TI - Phospholipase A(2)-catalyzed membrane leakage studied by immobilized liposome chromatography with online fluorescent detection. AB - Unilamellar liposomes composed of phosphatidylcholine with an entrapped self quenching fluorescent dye, calcein, were immobilized in chromatographic gel beads by avidin-biotin binding. Bee venom phospholipase A(2) (PLA(2)) was applied in a small amount onto the immobilized liposome column. The release of calcein from the immobilized liposomes resulting from the catalyzed hydrolysis of the phospholipids was detected online by immobilized liposome chromatography (ILC) using a flow fluorescent detector. The PLA(2)-catalyzed membrane leakage of the immobilized liposomes as studied with ILC was found to be affected by the gel pore size used for immobilization, by liposome size, and as expected by the concentration of calcium, but was unaffected by the flow rate of ILC. The largest PLA(2)-induced calcein release from the liposome column was detected on large unilamellar liposomes immobilized on TSK G6000PW or Sephacryl S-1000 gel in the presence of 1 mM Ca(2+) in the aqueous mobile phase. Comparison with the PLA(2) catalyzed membrane leakage in free liposome suspensions, we conclude that the fluorescent leakage from liposomes hydrolyzed by PLA(2) can be rapidly and sensitively detected by ILC runs using large amount of immobilized liposomes with entrapped fluorescent dye. PMID- 11399041 TI - A high-throughput screen for identifying transmembrane pore-forming peptides. AB - We have developed a visual microwell plate assay for rapid, high-throughput screening for membrane-disrupting molecules such as de novo designed pore formers, antibiotic peptides, bacterial toxins, and lipases. The detectability is based on the strong fluorescence emission of the lanthanide metal terbium(III) (Tb(3+)) when it interacts with the aromatic chelator dipicolinic acid (DPA). While Tb(3+) is not strongly fluorescent alone, the binary complex emits bright green fluorescence when irradiated with uv light. For the microwell plate assay, we prepared unilamellar phospholipid vesicles that had either Tb(3+) or DPA entrapped and the opposite molecule in the external solution. Disruption of the membranes allows the Tb(3+)/DPA complex to form, giving rise to a visibly fluorescent solution. In plates with 20-microl wells, the lower limit of visual detectability of the Tb(3+)/DPA complex in solution was about 2.5 microM. The lower limit of detectability using vesicles with entrapped Tb(3+) or DPA was about 50 microM phospholipid. We show that the membrane-disrupting effect of as little as 0.25 microM or 5 pmol of the pore-forming, antibiotic peptide alamethicin can be detected visually with this system. This sensitive, high throughput assay is readily automatable and makes possible the visual screening of combinatorial peptide libraries for members that permeabilize lipid bilayer membranes. PMID- 11399042 TI - A green fluorescent protein kinase substrate allowing detection and localization of intracellular ERK/MAP kinase activity. AB - We describe a versatile intracellular reporter of ERK/MAP kinase activity: a cDNA construct, pGFP.MBP, encoding amino acids 85-144 of the human myelin basic protein fused to the C-terminus of an enhanced green fluorescent protein (GFP). The fused fragment of myelin basic protein contains a single consensus ERK/MAP kinase phosphorylation motif (PRTP, where the threonine is phosphorylated). Phosphorylation of the specific motif can be detected via immunoblotting or immunofluorescence with a commercially available phospho-specific monoclonal antibody. When expressed in mammalian cells by either transient or stable transfection, the fusion protein acts as a bona fide kinase substrate, as demonstrated by rapid serum-induced phosphorylation that is blocked by a specific MEK inhibitor. Moreover, the localization of the total substrate pool is easily visualized by GFP autofluorescence and the extent of its phosphorylation simultaneously detected within intact fixed cells by immunofluorescence using the commercially available phospho-specific antibody. The approach described should be generally applicable to the intracellular analysis of many specific protein kinase substrates for which phospho-specific antibodies have been produced. PMID- 11399043 TI - A high-throughput assay for mitochondrial membrane potential in permeabilized yeast cells. AB - A fluorometric assay for mitochondrial membrane potential in permeabilized yeast cells has been developed. This method involves permeabilizing the plasma membrane and measuring the distribution of a mitochondrial membrane potential sensitive probe 3,3'-dipropylthiadicarbocyanine iodide (DiSC(3)(5); DiSC(3)). In permeabilized cells, DiSC(3) fluorescence decreased when introduced into energized mitochondria and increased three- to sixfold when the mitochondrial membrane potential was dissipated by the chemical uncoupler carbonylcyanide m chlorophenyl hydrazone. Plasma membrane potential was abolished by permeabilization, as shown by a lack of polarization of the plasma membrane induced by K(+) and glucose. Uncoupling protein 1 (UCP1), a mitochondrial H(+) transporter, was used as a model for method validation. The fluorescence intensity responded vigorously to specific modulators in UCP1-expressing cells. This method has been adapted as a high-throughput assay to screen for modulators of mitochondrial membrane potential. PMID- 11399044 TI - A homogeneous fluorometric assay for measuring cell adhesion to immobilized ligand using V-well microtiter plates. AB - We have developed a homogeneous high-capacity assay format for measuring integrin and selectin-dependent cell binding to immobilized ligand using V-well microtiter plates. 2',7'-Bis(2-carboxyethyl)-5-(and-6)-carboxylfluorescence, acetoxymethylester-labeled cells are added to ligand-coated V-shaped microtiter wells. Bound cells are separated from free cells using centrifugal force to produce shear stress. Nonadherent cells accumulate in the nadir of the well and are measured using a fluorescence plate reader. Antibody or low-molecular-weight inhibitors of either the ligand or the cell surface receptor result in less cell binding, more cells in the pellet, and increased signal. The optimization and validation of the very late antigen-4/vascular cell adhesion molecule-1 assay is described in detail. We demonstrate that this assay can be rapidly adapted to measure other integrin- and selectin-mediated interactions. This assay format has several advantages over conventional assays. The centrifugal process is biologically relevant and eliminates the washing steps to remove nonadherent cells that can cause well-to-well and plate-to-plate variation. Because the assay is robust with a high signal-to-noise ratio and low variability, it is ideally suited for studying multiple parameters of cell adhesion and for high capacity screening. PMID- 11399045 TI - Rapid separation of large DNA fragments. PMID- 11399046 TI - Hapten-labeled hyaluronan, a substrate to monitor hyaluronidase activity by enhanced chemiluminescence-assisted detection on filter blots. PMID- 11399047 TI - Effects of polymer dilution on quantitative polymerase chain reaction performed by capillary electrophoresis. PMID- 11399048 TI - Nitric oxide suppresses IL-8 transcription by inhibiting c-Jun N-terminal kinase induced AP-1 activation. AB - The role of activator protein-1 (AP-1) in tumor necrosis factor-alpha (TNF-alpha) induced interleukin-8 (IL-8) gene expression was evaluated. We showed that TNF alpha activates AP-1 in the transformed endothelial cell line ECV304 by transient transfections of IL-8 promoter construct pGL-3BF(2). Mutation of either the AP-1 site or the NF-IL-6 site on the IL-8 promoter suppressed the TNF-alpha-induced activation, suggesting cooperation between these transcription factors and transcription factor NF-kappaB. Overexpression of dominant negative mutants of c Jun suppressed AP-1-driven transcription of the IL-8 promoter following stimulation by TNF-alpha, suggesting that cooperative interaction between AP-1 and NF-kappaB is essential for IL-8 transcription in the presence of TNF-alpha. We also showed that nitric oxide (NO), in the form of an exogenous NO donor, suppressed the level of activation of the AP-1 subunit, c-Jun, by down-regulation of c-Jun NH2 terminal kinase. This down-regulation could be the putative mechanism of action for NO-mediated inhibition of IL-8 secretion in activated endothelium. These observations suggest for the first time that NO has broad suppressive activities on various proinflammatory effectors in activated endothelium. PMID- 11399049 TI - Regulation of the levels of small heat-shock proteins during differentiation of C2C12 cells. AB - Levels of the small heat-shock proteins (sHSPs) HSP27 and alphaB-crystallin during differentiation of mouse C2C12 cells were determined using specific immunoassays. Increases of these proteins were about 3-fold and 10-fold, respectively. Under the same conditions, however, the level of HSP70 in C2C12 cells barely increased, indicating selective accumulation of HSP27 and alphaB crystallin with differentiation. While expression of mRNA for alphaB-crystallin was also markedly increased and that for HSP27 was but to a lesser extent, mRNA for HSP70 could barely be detected during differentiation. Activation of the heat shock factor was not observed, in contrast to the case with heat-stressed undifferentiated cells. Various inhibitors of protein kinases affected the differentiation and the associated increase of sHSPs. Rapamycin, an inhibitor of p70 S6 kinase, completely inhibited the differentiation and suppressed the accumulation of HSP27 and alphaB-crystallin. SB203580, an inhibitor of p38 MAP kinase, also inhibited differentiation, but the accumulation of alphaB-crystallin was rather enhanced. PD98059, an inhibitor of MAP kinase kinase, significantly increased expression of a differentiation marker for muscle cells, creatine kinase M isozyme, as well as accumulation of alphaB-crystallin. These results suggest that accumulation of sHSPs during differentiation of C2C12 cells is regulated in a complex manner. PMID- 11399050 TI - Inhibition of neutrophil spreading during adhesion to fibronectin reveals formation of long tubulovesicular cell extensions (cytonemes). AB - Human neutrophils developed long thin tubulovesicular extensions (cytonemes) upon adhesion to fibronectin-coated substrata, when spreading was blocked. We observed extension formation when neutrophils were plated to fibronectin-coated substrata in Na(+)-free extracellular medium or in the presence of drugs capable of inhibiting spreading: 4-bromophenacyl bromide, N-ethylmaleimide, 7-chloro-4 nitrobenz-2-oxa-1,3-diazole, and cytochalasin D. Addition of Na(+) ions or washing of inhibitors restored neutrophil spreading. Phase-contrast and scanning electron microscopy revealed two types of extensions: (1) highly dynamic, flexible tubulovesicular extensions with unattached tips 0.2-0.4 microm in diameter, which can achieve 70-80 microm in length during 20 min, and (2) thinner straight extensions with flattened tips, which were formed in the presence of phorbol 12-myristate 13-acetate and connected cells to substratum or to the neighboring cells several cell diameters away. The latter may have derived from the former through tension after attachment of the tips. Spreading and extension formation may represent two states of the cell adhesive and communicative mechanism. PMID- 11399051 TI - Association of caveolin with Chlamydia trachomatis inclusions at early and late stages of infection. AB - The mechanism by which the intracellular bacterial pathogen Chlamydia trachomatis enters eukaryotic cells is poorly understood. There are conflicting reports of entry occurring by clathrin-dependent and clathrin-independent processes. We report here that C. trachomatis serovar K enters HEp-2 and HeLa 229 epithelial cells and J-774A.1 mouse macrophage/monocyte cells via caveolin-containing sphingolipid and cholesterol-enriched raft microdomains in the host cell plasma membranes. First, filipin and nystatin, drugs that specifically disrupt raft function by cholesterol chelation, each impaired entry of C. trachomatis serovar K. In control experiments, filipin did not impair entry of the same organism by an antibody-mediated opsonic process, nor did it impair entry of BSA-coated microspheres. Second, the chlamydia-containing endocytic vesicles specifically reacted with antisera against the caveolae marker protein caveolin. These vesicles are known to become the inclusions in which parasite replication occurs. They avoid fusion with lysosomes and instead traffic to the Golgi region, where they intercept Golgi-derived vesicles that recycle sphingolipids and cholesterol to the plasma membrane. We also report that late-stage C. trachomatis inclusions continue to display high levels of caveolin, which they likely acquire from the exocytic Golgi vesicles. We suggest that the atypical raft-mediated entry process may have important consequences for the host-pathogen interaction well after entry has occurred. These consequences include enabling the chlamydial vesicle to avoid acidification and fusion with lysosomes, to traffic to the Golgi region, and to intercept sphingolipid-containing vesicles from the Golgi. PMID- 11399052 TI - Transforming growth factor-beta1 promotes invasiveness after cellular transformation with activated Ras in intestinal epithelial cells. AB - Invasion is a defining event in carcinoma progression. In general, invasive carcinoma is characterized by an epithelial-fibroblastoid conversion associated with loss of cell-cell adhesion receptors such as E-cadherin and beta-catenin. We report here that TGF-beta1 promotes the invasiveness by modulating the alterations of cellular plasticity including a loss of cell-cell contact in Ras transformed epithelial cells. In order to examine the role of TGF-beta1 in the Ras-induced responses, intestinal epithelial cells expressing a conditionally activated Ha-Ras(Val12) (RIE-iRas cells) were used in this study. Induced expression of activated Ha-Ras(Val12) caused morphologic transformation of the RIE-iRas cells with an increase in vimentin expression and a decrease of E cadherin levels. There was also redistribution of beta-catenin from the cytoplasm to the nucleus after the induction of Ras. TGF-beta1 treatment enhanced both the decrease in E-cadherin levels and the redistribution of beta-catenin. Interestingly, the activation of Ras markedly decreased the level of TGF-beta receptor type II (TbetaRII) in RIE-iRas cells. However, the expression of plasminogen activator inhibitor-1, which is known to be transcriptionally induced by TGF-beta1, was strongly induced by TGF-beta1 despite the marked downregulation of TbetaRII. The induction of Ha-Ras(Val12) markedly increased the invasiveness in RIE-iRas cells, as evaluated by a collagen type I-coated Boyden-chamber assay, and the Ras-mediated invasiveness was significantly enhanced by TGF-beta1 treatment. Expression of a dominant-negative form of TbetaRII in the RIE-iRas cells abrogated both growth-inhibitory and invasion responses to TGF-beta1. Collectively, these results suggest that TGF-beta1 and oncogenic Ras collaborate in promoting cellular invasiveness in intestinal epithelial cells. The enhancement of invasiveness was correlated with decreased E-cadherin levels and subcellular distribution of beta-catenin. The enhancement of oncogenic Ras mediated cell transformation by TGF-beta1 occurs via TbetaRII. PMID- 11399053 TI - Expression of the helix-loop-helix protein ID1 in keratinocytes is upregulated by loss of cell-matrix contact. AB - To analyze the inhibitor of DNA-binding type 1 (ID1) in the human epidermis and in cultured keratinocytes we generated and characterized ID1-specific monoclonal antibodies. Immunohistological studies on human skin biopsies revealed that ID1 is not detectable in normal human epidermis but in lesional epidermis of bullous pemphigoid. In the latter case we found ID1 in the cytoplasm of basal and proximal suprabasal keratinocytes. Cultured normal human epidermal keratinocytes displayed ID1 in the cytoplasm; upon differentiation into a multilayered keratinocyte sheet, ID1 was no longer detectable. It was reexpressed after dispase-mediated detachment of the keratinocyte cultures from the growth substratum. In this case ID1 was localized to the cytoplasm and the nucleus. Our data indicate that after epidermal injury-in our case loss of cell-matrix contact ID1 is upregulated in affected keratinocytes. In view of the ID1 function in other cell types, we speculate that ID1 facilitates the transition from the resting to the migrating and proliferating keratinocyte required for efficient repair of epidermal lesions by reepithelialization. Taken together we suggest that ID1 is an important player in epidermal (patho-)physiology. PMID- 11399054 TI - Peroxisomes exist in growth cones and move anterogradely and retrogradely in neurites of PC12D cells. AB - Localization and movement of peroxisomes have been investigated in neurites of a subline of PC12 pheochromocytoma cells (PC12D cells). The cells were transfected with a construct encoding the green fluorescent protein and bearing the C terminal peroxisomal targeting signal 1 SKL motif (-Ser-Lys-Leu-COOH). Peroxisomes were detected as green punctate fluorescent signals. Many peroxisomes were observed in neurites of PC12D cells, especially in neural terminal-like structures, growth cones, varicosities, and branch points. Growth cones containing many peroxisomes were active, since they extended several long filopodias. Existence of peroxisomes in growth cones and neuronal terminal-like structures suggests that peroxisomes might have some role in neuronal extension and nerve terminal functioning. Peroxisomal motility was analyzed by time-lapse imaging using a fluorescence microscope at 25 degrees C. Peroxisomes were transported bidirectionally in neurites, i.e., through anterograde and retrograde transport. This result suggests that peroxisomes move to growth cones and neural terminals from the PC12D cell body, play some role in these parts, and go back to cell body. PMID- 11399055 TI - Cell cycle-dependent changes in localization of a 210-kDa microtubule-interacting protein in Leishmania. AB - Using the monoclonal antibody MA-01, a new 210-kDa microtubule-interacting protein was identified in Leishmania promastigotes by immunoblotting and by immunoprecipitation. The protein was thermostable and was located on microtubules prepared by taxol-driven polymerization in vitro. On fixed cells the antibody gave specific staining of flagellum, flagellar pocket, and mitotic spindle. Subpellicular microtubules were basically not decorated but posterior poles of the cells were labeled in a cell-cycle-dependent manner. In anterior and posterior poles of cells the 210-kDa protein codistributed with the 57-kDa protein, immunodetected with anti-vimentin antibody, that was located only on cell poles. Immunolocalization of the 57-kDa protein was most prominent in dividing cells. The presented data suggest that the 210-kDa protein is a newly identified microtubule-interacting protein of Leishmania that could be involved in anchoring the microtubules in posterior poles of these cells. The striking codistribution of the microtubule-interacting protein and the 57-kDa protein in protozoa is described for the first time. PMID- 11399056 TI - Caspase-3 is actively involved in okadaic acid-induced lens epithelial cell apoptosis. AB - Phosphorylation and dephosphorylation are important cellular events regulating major metabolic activities such as signal transduction, gene expression, cell cycle progression, and apoptosis. It is well documented that okadaic acid, a potent inhibitor of protein phosphatase-1 (PP-1) and -2A (PP-2A), can induce apoptosis in a variety of cell lines. Our recent studies have revealed that in the immortal rabbit lens epithelial cell line, N/N1003A, inhibition of PP-1, but not PP-2A, leads to rapid apoptosis of the lens epithelial cells. This induction of cell death is associated with up-regulated expression of a set of genes, including the tumor-suppressor gene, p53, and the proapoptotic gene, bax. In the present study, we demonstrate that inhibition of PP-1 by okadaic acid in the primary cultures of rat lens epithelial cells also leads to apoptotic death. Moreover, we show that the cysteine protease, caspase-3, is important in the execution of okadaic acid-induced apoptosis. Treatment of the primary cultures of rat lens epithelial cells with 100 nM okadaic acid up-regulates expression of caspase-3 at the mRNA, protein, and enzyme activity levels. Inhibition of the caspase-3 activity with a chemically synthesized inhibitor prevents okadaic acid induced apoptosis in rat lens epithelial cells. Similar results are also observed in the immortal cell line N/N1003A. Furthermore, stable expression of the mouse gene encoding lens alphaB crystallin inhibits okadaic acid-induced apoptosis, and this inhibition is associated with repression of the okadaic acid-induced up regulation of caspase-3 activity. Taken together, these results demonstrate that caspase-3 is actively involved in okadaic acid-induced lens epithelial cell apoptosis. PMID- 11399057 TI - Y-27632, an inhibitor of Rho-associated kinases, prevents tyrosine phosphorylation of focal adhesion kinase and paxillin induced by bombesin: dissociation from tyrosine phosphorylation of p130(CAS). AB - A rapid increase in tyrosine phosphorylation of focal adhesion kinase (FAK), paxillin, and Crk-associated substrate (CAS) are prominent early events triggered by many G protein-coupled receptors (GPCRs), but the mechanisms involved remain unclear. Here, we examined whether the Rho-associated protein serine/threonine kinase family (ROCK) is a critical Rho effector in the pathway that links GPCR activation to the tyrosine phosphorylation of FAK, CAS, and paxillin. Treatment of Swiss 3T3 cells with Y-27632, a preferential inhibitor of ROCK, dramatically inhibited the formation of actin stress fibers, the assembly of focal contacts, and the increase in tyrosine phosphorylation of FAK and paxillin induced by bombesin in these cells. Surprisingly, we found that treatment with Y-27632 did not produce any detectable effect on bombesin-elicited CAS tyrosine phosphorylation even at the highest concentrations of Y-27632 tested. HA-1077, a preferential inhibitor of ROCK activity structurally unrelated to Y-27632, also attenuated the increase in the tyrosine phosphorylation of FAK and paxillin but did not affect the tyrosine phosphorylation of CAS induced by bombesin in Swiss 3T3 cells. The results demonstrate that ROCK-dependent tyrosine phosphorylation of FAK and paxillin can be dissociated from a ROCK-independent pathway leading to tyrosine phosphorylation of CAS. PMID- 11399058 TI - Accumulation of advanced glycation end products decreases collagen turnover by bovine chondrocytes. AB - The integrity of the collagen network is essential for articular cartilage to fulfill its function in load support and distribution. Damage to the collagen network is one of the first characteristics of osteoarthritis. Since extensive collagen damage is considered irreversible, it is crucial that chondrocytes maintain a functional collagen network. We investigated the effects of advanced glycation end products (AGEs) on the turnover of collagen by articular cartilage chondrocytes. Increased AGE levels (by culturing in the presence of ribose) resulted in decreased collagen synthesis (P < 0.05) and decreased MMP-mediated collagen degradation (P < 0.02). The latter could be attributed to increased resistance of the collagen network to MMPs (P < 0.05) as well as the decreased production of MMPs by chondrocytes (P < 0.02). Turnover of a protein is determined by its synthesis and degradation rates and therefore these data indicate that collagen turnover is decreased at enhanced AGE levels. Since AGE levels in human cartilage increase approximately 50 fold between age 20 and 80, cartilage collagen turnover likely decreases with increasing age. Impaired collagen turnover adversely affects the capacity of chondrocytes to remodel and/or repair its extracellular matrix. Consequently, age-related accumulation of AGE (via decreased collagen turnover) may contribute to the development of cartilage damage in osteoarthritis. PMID- 11399059 TI - Role of stem cell factor and c-kit signaling in regulation of fetal intestinal epithelial cell adhesion to fibronectin. AB - The interaction of stem cell factor (SCF) and c-kit is considered to be an important signaling event for the homeostasis of the epithelial barrier function in the intestinal tract. This study was designed to investigate the role of the SCF and c-kit signaling pathway in adhesion of intestinal epithelial cells (IECs) to fibronectin (FN) using primary cells. Fetal murine IECs were prepared from the small intestine of mouse fetus. The mRNAs coding for SCF in mesenchymes and c-kit in IECs were detected by reverse transcription-PCR. The expression of FN receptor VLA-5 on IECs was examined by flow cytometry. A cell adhesion assay showed that the stimulation of IECs with SCF increased the number of cells adhering to FN. Experiments using specific antibody against SCF indicated that this increase in cell adhesion was SCF-dependent. On the other hand, SCF did not influence the expression of VLA-5 on IECs. The IEC adhesion to FN was inhibited by specific antibody against the FN receptor (VLA-5), as well as competitive Arg-Gly-Asp (RGD) peptide. When alteration of intracellular signal transduction induced by SCF was examined, it was found that SCF stimulated a tyrosine-specific c-kit autophosphorylation cascade of IECs. Further, preincubation of IECs with an optimal concentration of genistein resulted in the inhibition of SCF-induced c kit phosphorylation and adhesion of IECs to FN. These results suggested that adhesion of immature IECs to FN is regulated by activation of RGD-dependent VLA-5 through the SCF and c-kit signal transduction pathway. SCF, which may be produced by mesenchymes locally, is an important regulatory factor for the adhesion of immature IECs to basement membrane matrix via VLA-5 and FN interaction. This cytokine-regulated interaction between VLA-5 and FN may play an important role in the development and wound repair of the intestinal tract. PMID- 11399060 TI - Modulation of caspase-3 activity by zinc ions and by the cell redox state. AB - It is known that DNA fragmentation during apoptosis is controlled by a number of factors, a crucial step being the caspase-operated cleavage of ICAD, the DNase inhibitor. We have previously demonstrated that hydrogen peroxide-treated lymphocytes undergo apoptosis without formation of a DNA ladder; however, the use of micromolar amounts of a Zn(2+) chelator allowed DNA cleavage at internucleosomal sites. Such results were extended in the present work, thus allowing their framing into the events related to alterations in the redox state of the cell. Apoptosis in hydrogen peroxide-treated lymphocytes was found to occur with caspase-3 activation, but the enzyme activity was found to be impaired, thus affecting internucleosomal fragmentation as well as nuclear morphology. Caspase-3 activity was found to resume upon mild Zn(2+) chelation. These results provide as well an experimental model from which apoptotic events upstream and downstream of caspase-3 activity can be examined. PMID- 11399061 TI - Different roles of spermine in glucocorticoid- and Fas-induced apoptosis. AB - Two experimental systems representative of the mitochondrial and death receptor apoptotic pathways are the dexamethasone-induced programmed cell death in mouse thymocytes and the antibody-mediated cross-ligation of the Fas receptor in the human leukemic T-cell line Jurkat, respectively. In both cell systems, caspase-9, -8, and -3 were activated upon induction of apoptosis and a sub-G(1) peak appeared as a sign of ongoing DNA fragmentation. Addition of 1 mM spermine together with dexamethasone inhibited caspase activation and the appearance of the sub-G(1) peak in mouse thymocytes. In contrast, Fas-induced cell death was totally unaffected by spermine addition. Spermine addition significantly elevated the spermine concentration in both thymocytes and Jurkat cells. Thus, spermine per se did not inhibit the caspases but rather their activation. The fact that spermine inhibited caspase activation only in the thymocytes implies that spermine inhibited dexamethasone-induced apoptosis upstream of caspase-9 activation. PMID- 11399062 TI - Protein surface amino acid compositions distinctively differ between thermophilic and mesophilic bacteria. AB - One of the well-known observations of proteins from thermophilic bacteria is the bias of the amino acid composition in which charged residues are present in large numbers, and polar residues are scarce. On the other hand, it has been reported that the molecular surfaces of proteins are adapted to their subcellular locations, in terms of the amino acid composition. Thus, it would be reasonable to expect that the differences in the amino acid compositions between proteins of thermophilic and mesophilic bacteria would be much greater on the protein surface than in the interior. We performed systematic comparisons between proteins from thermophilic bacteria and mesophilic bacteria, in terms of the amino acid composition of the protein surface and the interior, as well as the entire amino acid chains, by using sequence information from the genome projects. The biased amino acid composition of thermophilic proteins was confirmed, and the differences from those of mesophilic proteins were most obvious in the compositions of the protein surface. In contrast to the surface composition, the interior composition was not distinctive between the thermophilic and mesophilic proteins. The frequency of the amino acid pairs that are closely located in the space was also analyzed to show the same trend of the single amino acid compositions. Interestingly, extracellular proteins from mesophilic bacteria showed an inverse trend against thermophilic proteins (i.e. a reduced number of charged residues and rich in polar residues). Nuclear proteins from eukaryotes, which are known to be abundant in positive charges, showed different compositions as a whole from the thermophiles. These results suggest that the bias of the amino acid composition of thermophilic proteins is due to the residues on the protein surfaces, which may be constrained by the extreme environment. PMID- 11399063 TI - The crystal structure of delta(3)-delta(2)-enoyl-CoA isomerase. AB - The active-site geometry of the first crystal structure of a Delta(3)-Delta(2) enoyl-coenzyme A (CoA) isomerase (the peroxisomal enzyme from the yeast Saccharomyces cerevisiae) shows that only one catalytic base, Glu158, is involved in shuttling the proton from the C2 carbon atom of the substrate, Delta(3)-enoyl CoA, to the C4 atom of the product, Delta(2)-enoyl-CoA. Site-directed mutagenesis has been performed to confirm that this glutamate residue is essential for catalysis. This Delta(3)-Delta(2)-enoyl-CoA isomerase is a hexameric enzyme, consisting of six identical subunits. It belongs to the hydratase/isomerase superfamily of enzymes which catalyze a wide range of CoA-dependent reactions. The members of the hydratase/ isomerase superfamily have only a low level of sequence identity. Comparison of the crystal structure of the Delta(3)-Delta(2) enoyl-CoA isomerase with the other structures of this superfamily shows only one region of large structural variability, which is in the second turn of the spiral fold and which is involved in defining the shape of the binding pocket. PMID- 11399064 TI - Autoregulation enables different pathways to control CCAAT/enhancer binding protein beta (C/EBP beta) transcription. AB - CCAAT/enhancer binding protein beta (C/EBP beta) also named liver-enriched transcriptional activating protein (LAP) is a member of the C/EBP family of transcription factors and is involved in hepatocyte-specific gene expression and in the process of tissue differentiation. The activity of LAP/C/EBP beta can be regulated at the transcriptional and posttranslational level or by protein protein interaction with other transcription factors. In this study we show that LAP/C/EBP beta can stimulate its own transcription. Deletion analysis of the rat LAP/C/EBP beta promoter in luciferase reporter gene experiments demonstrated that the region located between nucleotide -121 to -71, comprising two recently characterized cAMP responsive element (CRE)-like elements, is important for autoregulation. Gel shift experiments using oligonucleotides with overlapping point mutations identified the sequence GCAATGA (beta-site) adjacent to and partially overlapping the first CRE-like site as core motif for LAP/C/EBP beta binding. Analysis of a mutated beta-site in reporter gene experiments showed the functional relevance of this site for autoregulation. The composite C/EBP beta CRE-element in the promoter enables synergistic activation of transcription by LAP/C/EBP beta and the protein kinase A (PKA)/cAMP responsive element binding protein (CREB) pathway in a cell-type specific manner. In hepatoma cells nuclear factor kappa B (NF-kappa B) increased autoregulation and therefore could mediate enhanced activation during inflammatory responses. In summary, our results demonstrated that the assembly of the three binding sites in the promoter and thus the interaction between LAP/C/EBP beta and members of the CREB or NF-kappa B family allows the control of LAP/C/EBP beta gene transcription as a response to different stimuli in a tissue specific manner. PMID- 11399065 TI - Dissection of the ATP-driven reaction cycle of the bacteriophage T4 DNA replication processivity clamp loading system. AB - Processive DNA replication requires the loading of a multisubunit ring-shaped protein complex, known as a sliding or processivity clamp, onto the primer template (p/t) DNA. This clamp then binds to the replication polymerase to form a processive polymerase holoenzyme. The processivity of the holoenzyme derives from the topological properties of the clamp, which encircles the DNA without actually binding to it. Multisubunit complexes known as clamp-loaders utilize ATP to drive the placement of this ring around the DNA. To further understand the role of ATP binding and hydrolysis in driving clamp-loading in the DNA replication system of bacteriophage T4, we report the results of a series of presteady-state and steady state kinetic ATPase experiments involving the various components of the reconstituted system. The results obtained are consistent with a mechanism in which a slow step, which involves the binary ATP-bound clamp-clamp loader complex, activates this complex and permits p/t DNA to bind and stimulate ATP hydrolysis. ATP hydrolysis itself, as well as the subsequent (after clamp loading) dissociation of the clamp-loader and the slippage of the loaded clamp from the p/t DNA construct, are shown to be fast steps. A second slow step occurs after ATP hydrolysis. This step involves the dissociated clamp loader complex and may reflect ADP release. Only one molecule of ATP is hydrolyzed per clamp-loading event. Rate constants for each step, and an overall reaction mechanism for the T4 clamp-loading system, are derived from these data and from other results in the literature. The principles that emerge fit into a general framework that can apply to many biological processes involving ATP-driven reaction cycles. PMID- 11399066 TI - Hairpin ribozymes with four-way helical junctions mediate intracellular RNA ligation. AB - Virtually all RNA-mediated reactions require transitions among alternative RNA conformations. The complexity of biological reactions can obscure specific conformational changes in vivo and important features of the intracellular environment are difficult to reproduce in vitro. However, simple RNA self cleavage and ligation reactions offer a unique opportunity to measure the kinetics and equilibria of specific RNA conformational transitions directly in living cells. Hairpin ribozymes that incorporate the natural four-way helical junction self-cleave rapidly in vivo, but only when cleavage products dissociate rapidly. Cleavage rates fall when cleavage products remain bound in stable base paired helices, providing evidence that bound products undergo re-ligation. These results provide the first detailed kinetic description of an intracellular ribozyme reaction that includes cleavage, ligation and product dissociation rates. Kinetic and equilibrium parameters measured in vivo correspond well, but not perfectly, with values measured for the same reactions in vitro under conditions that approximate an intracellular ionic environment. PMID- 11399067 TI - Finding and using local symmetry in identifying lower domain movements in hexon subunits of the herpes simplex virus type 1 B capsid. AB - A characteristic of virus assembly is the use of symmetry to construct a complex capsid from a limited number of different proteins. Many spherical viruses display not only icosahedral symmetry, but also local symmetries, which further increase the redundancy of their structural proteins. We have developed a computational procedure for evaluating the quality of these local symmetries that allows us to probe the extent of local structural variations among subunits. This type of analysis can also provide orientation parameters for carrying out non icosahedral averaging of quasi-equivalent subunits during three-dimensional structural determination. We have used this procedure to analyze the three types of hexon (P, E and C) in the 8.5 A resolution map of the herpes simplex virus type 1 (HSV-1) B capsid, determined by electron cryomicroscopy. The comparison of the three hexons showed that they have good overall 6-fold symmetry and are almost identical throughout most of their lengths. The largest difference among the three lies near the inner surface in a region of about 34 A in thickness. In this region, the P hexon displays slightly lower 6-fold symmetry than the C and E hexons. More detailed analysis showed that parts of two of the P hexon subunits are displaced counterclockwise with respect to their expected 6-fold positions. The most highly displaced subunit interacts with a subunit from an adjacent P hexon (P'). Using the local 6-fold symmetry axis of the P hexon as a rotation axis, we examined the geometrical relationships among the local symmetry axes of the surrounding capsomeres. Deviations from exact symmetry are also found among these local symmetry axes. The relevance of these findings to the process of capsid assembly is considered. PMID- 11399068 TI - Crystal structure of a heptameric Sm-like protein complex from archaea: implications for the structure and evolution of snRNPs. AB - The Sm/Lsm proteins associate with small nuclear RNA to form the core of small nuclear ribonucleoproteins, required for processes as diverse as pre-mRNA splicing, mRNA degradation and telomere formation. The Lsm proteins from archaea are likely to represent the ancestral Sm/Lsm domain. Here, we present the crystal structure of the Lsm alpha protein from the thermophilic archaeon Methanobacterium thermoautotrophicum at 2.0 A resolution. The Lsm alpha protein crystallizes as a heptameric ring comprised of seven identical subunits interacting via beta-strand pairing and hydrophobic interactions. The heptamer can be viewed as a propeller-like structure in which each blade consists of a seven-stranded antiparallel beta-sheet formed from neighbouring subunits. There are seven slots on the inner surface of the heptamer ring, each of which is lined by Asp, Asn and Arg residues that are highly conserved in the Sm/Lsm sequences. These conserved slots are likely to form the RNA-binding site. In archaea, the gene encoding Lsm alpha is located next to the L37e ribosomal protein gene in a putative operon, suggesting a role for the Lsm alpha complex in ribosome function or biogenesis. PMID- 11399069 TI - Incorporation of beta-selenolo[3,2-b]pyrrolyl-alanine into proteins for phase determination in protein X-ray crystallography. AB - beta-Selenolo[3,2-b]pyrrolyl-L-alanine that mimics tryptophan with the benzene ring of the indole moiety replaced by selenophene, was incorporated into human annexin V and barstar. This was achieved by fermentation and expression in a Trp auxotrophic Escherichia coli host strain using the selective pressure incorporation method. The seleno- proteins were obtained in yields comparable to those of the wild-type proteins and exhibit full crystallographic isomorphism to the parent proteins, but expectedly show altered absorbance profiles and quenched tryptophan fluorescence. Since the occurrence of tryptophan residues in proteins is rare, incorporation of the electron-rich selenium-containing tryptophan surrogate into proteins represents a useful supplementation and even a promising novel alternative to selenomethionine for solving the phase problem in protein X ray crystallography. PMID- 11399070 TI - Domain closure mechanism in transferrins: new viewpoints about the hinge structure and motion as deduced from high resolution crystal structures of ovotransferrin N-lobe. AB - The crystal structure of holo hen ovotransferrin N-lobe refined at 1.65 A resolution has been obtained. The final model gave an R-factor of 0.173 in the resolution range between 10.0 and 1.65 A. The comparison of the structure with previous high-resolution apo and Fe(3+)-loaded, domain-opened intermediate structures provides new viewpoints on the domain closure mechanism upon Fe(3+) uptake in ovotransferrin N-lobe. Overall, conformational transition follows the common mechanism that has been first demonstrated for lactoferrin N-lobe; the domains 1 and 2 rotate 49.7 degrees as rigid bodies with a translation of 2.1 A around a screw-axis that passes through the two interdomain beta-strands (89-94 and 244-249). It is generally believed that the two strands display a hinge-like motion. Here, the latter strand indeed displays an ideal hinge nature: the segments 244-246 and 248-249 behave as a part of the rigid body of domain 2 and that of domain 1, respectively, and a sharp bend upon the domain closure is largely accounted for by the changes in the torsion angles phi and psi of Val247. We find, however, that the mode of the conformational change in the first beta strand is much more complex. Two of the five inter beta-strand hydrogen bonds undergo crucial exchanges: from Ser91-N...Val247-O and Thr89-O...Ala249-N in the open apo and intermediate structures into Tyr92-N...Val247-O and Thr90-O...Ala249 N in the closed holo structure. These exchanges, which may be triggered in the intermediate state by modulation in the topological relation between the Fe(3+) ligated hinge residue Tyr92-OH and the anion anchor residues of helix 5, are accompanied by a large conformational change and extensive hydrogen bonding rearrangements in a long stretch of segment of Glu82 to Tyr92. Such structural transition would work as a driving force for the domain closure, which highlights a "door closer"-like role, in addition to the canonical-hinge role, for the interdomain polypeptide segment pair. As an alternative hinge that secures the correct domain motion by being placed on a significant distance from the beta strand hinge, we point out the participation of the van der Waals contacts formed between domain 1 residue of Met331 and domain 2 residues of Trp125, Ile129 and Trp140. PMID- 11399071 TI - The solution structure of the N-terminal domain of riboflavin synthase. AB - The structure of the amino-terminal domain of Escherichia coli riboflavin synthase (RiSy) has been determined by NMR spectroscopy with riboflavin as a bound ligand. RiSy is functional as a 75 kDa homotrimer, each subunit of which consists of two domains which share very similar sequences and structures. The N terminal domain (RiSy-N; 97 residues) forms a 20 kDa homodimer in solution which binds riboflavin with high affinity. The structure features a six-stranded antiparallel beta-barrel with a Greek-key fold, both ends of which are closed by an alpha-helix. One riboflavin molecule is bound per monomer in a site at one end of the barrel which is comprised of elements of both monomers. The structure and ligand binding are similar to that of the FAD binding domains of ferrodoxin reductase family proteins. The structure provides insights into the structure of the whole enzyme, the organisation of the functional trimer and the mechanism of riboflavin synthesis. C48 from the N-terminal domain is identified as the free cysteine implicated in a nucleophilic role in the synthesis mechanism, while H102 from the C-terminal domains is also likely to play a key role. Both are invariant in all known riboflavin synthase sequences. PMID- 11399072 TI - Ligand-induced structural changes to maltodextrin-binding protein as studied by solution NMR spectroscopy. AB - Solution NMR studies on the physiologically relevant ligand-free and maltotriose bound states of maltodextrin-binding protein (MBP) are presented. Together with existing data on MBP in complex with beta-cyclodextrin (non-physiological, inactive ligand), these new results provide valuable information on changes in local structure, dynamics and global fold that occur upon ligand binding to this two-domain protein. By measuring a large number of different one-bond residual dipolar couplings, the domain conformations, critical for biological function, were investigated for all three states of MBP. Structural models of the solution conformation of MBP in a number of different forms were generated from the experimental dipolar coupling data and X-ray crystal structures using a quasi rigid-body domain orientation algorithm implemented in the structure calculation program CNS. Excellent agreement between relative domain orientations in ligand free and maltotriose-bound solution conformations and the corresponding crystal structures is observed. These results are in contrast to those obtained for the MBP/beta-cyclodextrin complex where the solution state is found to be approximately 10 degrees more closed than the crystalline state. The present study highlights the utility of residual dipolar couplings for orienting protein domains or macromolecules with respect to each other. PMID- 11399073 TI - Folding of beta-sheets in membranes: specificity and promiscuity in peptide model systems. AB - The interactions that drive the folding of beta-barrel membrane proteins have not been well studied because there have been few available model systems for membrane beta-sheets. In this work, we expand on a recently described model system to explore the contributions of interstrand hydrogen bonds, side chain/side-chain interactions and side-chain/membrane interactions to beta-sheet formation in membranes. These experiments are based on the observation that the hydrophobic hexapeptide acetyl-Trp-Leu-Leu-Leu-Leu-Leu-OH (AcWLLLLL) folds, cooperatively and reversibly, into oligomeric, antiparallel beta-sheets in phosphatidylcholine membranes. To systematically characterize the important interactions that drive beta-sheet formation in membranes, we have used circular dichroism spectroscopy to determine the membrane secondary structure of each member of a complete host-guest family of related peptides of the form AcWLL-X LL, where X is one of the natural amino acids. Peptides with hydrophobic X residues of any size or character (X=Ala, Val, Ile, Leu, Cys, Met, Phe and Trp) form similar beta-sheets in membranes, while peptides with any polar X-residue or Gly or Pro at the X-position are random-coils, even when bound to membranes at high concentrations. The observed membrane sheet preferences correlate poorly with intrinsic sheet propensity scales measured in soluble proteins, but they correlate well with several membrane hydrophobicity scales. These results support the idea that the predominant interactions of the side-chains in membrane-bound beta-sheets are with the membrane lipids, and that backbone hydrogen bonding is the major driving force for the stabilization of beta-sheets in membranes. PMID- 11399074 TI - A succession of substrate induced conformational changes ensures the amino acid specificity of Thermus thermophilus prolyl-tRNA synthetase: comparison with histidyl-tRNA synthetase. AB - We describe the recognition by Thermus thermophilus prolyl-tRNA synthetase (ProRSTT) of proline, ATP and prolyl-adenylate and the sequential conformational changes occurring when the substrates bind and the activated intermediate is formed. Proline and ATP binding cause respectively conformational changes in the proline binding loop and motif 2 loop. However formation of the activated intermediate is necessary for the final conformational ordering of a ten residue peptide ("ordering loop") close to the active site which would appear to be essential for functional tRNA 3' end binding. These induced fit conformational changes ensure that the enzyme is highly specific for proline activation and aminoacylation. We also present new structures of apo and AMP bound histidyl-tRNA synthetase (HisRS) from T. thermophilus which we compare to our previous structures of the histidine and histidyl-adenylate bound enzyme. Qualitatively, similar results to those observed with T. thermophilus prolyl-tRNA synthetase are found. However histidine binding is sufficient to induce the co-operative ordering of the topologically equivalent histidine binding loop and ordering loop. These two examples contrast with most other class II aminoacyl-tRNA synthetases whose pocket for the cognate amino acid side-chain is largely preformed. T. thermophilus prolyl-tRNA synthetase appears to be the second class II aminoacyl-tRNA synthetase, after HisRS, to use a positively charged amino acid instead of a divalent cation to catalyse the amino acid activation reaction. PMID- 11399075 TI - Sfl1 functions via the co-repressor Ssn6-Tup1 and the cAMP-dependent protein kinase Tpk2. AB - Ssn6 (Cyc8) is a component of the yeast general corepressor Ssn6-Tup1 that inhibits the transcription of many diversely regulated genes. The corepressor does not interact directly with DNA but is recruited to different promoters through interactions with distinct pathway-specific, DNA-binding repressor proteins. Using yeast two-hybrid and GST chromatography interaction experiments, we have determined that Sfl1, a novel repressor protein, interacts directly with Ssn6, and in vivo repression data suggest that Sfl1 inhibits transcription by recruiting Ssn6-Tup1 via a specific domain in the Sfl1 protein. Sin4 and Srb10, components of specific RNA polymerase II sub-complexes that are required for Ssn6 Tup1 repression activity, are found to be required for Sfl1 repression function. These results indicate a possible mechanism for Sfl1-mediated repression via Ssn6 Tup1 and specific subunits of the RNA polymerase II holoenzyme. Electrophoretic mobility shift and chromatin immuno-precipitation assays demonstrate that Sfl1 is present at the promoters of three Ssn6-Tup1-repressible genes; namely, FLO11, HSP26, and SUC2. Sfl1 is known to interact with Tpk2, a cAMP-dependent protein kinase that negatively regulates Sfl1 function. Consistently, we show that phosphorylation by protein kinase A inhibits Sfl1 DNA binding in vitro, and that a tpk2Delta mutation increases the levels of Sfl1 protein associated with specific promoter elements in vivo. These data indicate a possible mechanism for regulating Sfl1-mediated repression through modulation of DNA binding by cAMP dependent protein kinase-dependent phosphorylation. Taken together with previous data, these new observations suggest a link between cAMP signaling and Ssn6-Tup1 mediated transcriptional repression. PMID- 11399076 TI - Differential expression of two plant-like enolases with distinct enzymatic and antigenic properties during stage conversion of the protozoan parasite Toxoplasma gondii. AB - The precise molecular mechanisms underlying the switch between the two developmental stages of Toxoplasma gondii, and the metabolic adaptations occurring during this stage conversion are poorly understood. Because inhibitors of mitochondrial respiration are known to trigger differentiation from tachyzoite into bradyzoite stages, we believe that some of the switch components may be sought in the regulation of central carbohydrate metabolism. We have previously described a cDNA encoding a bradyzoite-specific enolase, ENO1. We now report the isolation and characterization of another enolase-encoding cDNA (ENO2) that is expressed preferentially in the tachyzoite stage. The deduced amino acid sequences of ENO1 and ENO2 share 73.65 % identity. They both display significant homologies to plant enolases with the presence of two plant-like peptide insertions, a pentapeptide EWGW(Y)C(S) and a dipeptide EK (or DK). We demonstrate that deletions of the ENO1 pentapeptide motif on its own or together with the dipeptide reduce drastically the affinity for the 2PGA substrate, suggesting that the evolutionary acquisition of these peptides in enolases of land plants and apicomplexan parasites contribute a specific function to their enzymatic activities. T. gondii ENO1 and ENO2 were also expressed as active recombinant enzymes in Escherichia coli. While ENO1 and ENO2 display similar K(m) values, the pure tachyzoite-specific enzyme (ENO2) has a threefold specific activity at V(max) compared with that of the bradyzoite-specific enolase (ENO1). Moreover, immunoblot analyses performed using polyclonal antibodies raised against the recombinant enzymes revealed that the native enolase in tachyzoite and bradyzoite are also antigenically distinct. Taken together, our results indicate that the differences witnessed between the two activities may be instrumental in maintaining glycolysis in pace with the distinct stage-specific requirements of carbohydrate metabolism. PMID- 11399077 TI - Analysis of the roles of tRNA structure, ribosomal protein L9, and the bacteriophage T4 gene 60 bypassing signals during ribosome slippage on mRNA. AB - A 50-nucleotide coding gap divides bacteriophage T4 gene 60 into two open reading frames. In response to cis-acting stimulatory signals encrypted in the mRNA, the anticodon of the ribosome-bound peptidyl tRNA dissociates from a GGA codon at the end of the first open reading frame and pairs with a GGA codon 47 nucleotides downstream just before the second open reading frame. Mutations affecting ribosomal protein L9 or tRNA(Gly)(2), the tRNA that decodes GGA, alter the efficiency of bypassing. To understand the mechanism of ribosome slippage, this work analyzes the influence of these bypassing signals and mutant translational components on -1 frameshifting at G GGA and hopping over a stop codon immediately flanked by two GGA glycine codons (stop-hopping). Mutant variants of tRNA(Gly)(2) that impair bypassing mediate stop-hopping with unexpected landing specificities, suggesting that these variants are defective in ribosomal P-site codon-anticodon pairing. In a direct competition between -1 frameshifting and stop-hopping, the absence of L9 promotes stop-hopping at the expense of -1 frameshifting without substantially impairing the ability of mutant tRNA(Gly)(2) variants to re-pair with the mRNA by sub-optimal pairing. These observations suggest that L9 defects may stimulate ribosome slippage by enhancing mRNA movement through the ribosome rather than by inducing an extended pause in translation or by destabilizing P site pairing. Two of the bypassing signals, a cis-acting nascent peptide encoded by the first open reading frame and a stemloop signal located in the 5' portion of the coding gap, stimulate peptidyl-tRNA slippage independently of the rest of the gene 60 context. Evidence is presented suggesting that the nascent peptide signal may stimulate bypassing by destabilizing P-site pairing. PMID- 11399078 TI - Birth of a gene: locus of neuronal BC200 snmRNA in three prosimians and human BC200 pseudogenes as archives of change in the Anthropoidea lineage. AB - The gene encoding brain-specific dendritic BC200 small non-messenger RNA is limited to the primate order and arose from a monomeric Alu element. It is present and neuronally expressed in all Anthropoidea examined. By comparing the human sequence of about 13.2 kb with each of the prosimian (lemur 14.6 kb, galago 12 kb, and tarsier 13.8 kb) orthologous loci, we could establish that the BC200 RNA gene is absent from the prosimian lineages. In Strepsirhini (lemurs and lorises), a dimeric AluJ-like element integrated very close to the BC200 insertion point, while the corresponding tarsier region is devoid of any repetitive element. Consequently, insertion of the Alu monomer that gave rise to the BC200 RNA gene must have occurred after the anthropoid lineage diverged from the prosimian lineage(s). Shared insertions of other repetitive elements favor proximity of simians and tarsiers in support of their grouping into Haplorhini and the omomyid hypothesis. On the other hand, the nucleotide sequences in the segment that is available for comparison in all four species reveal less exchanges between Strepsirhini (lemur and galago) and human than between tarsier and human. Our data imply that the early activity of dimeric Alu sequences must have been concurrent with the activity of monomeric Alu elements that persisted longer than is usually thought. As BC200 RNA gave rise to more than 200 pseudogenes, we used their consensus sequence variations as a molecular archive recording the BC200 RNA sequence changes in the anthropoid lineage leading to Homo sapiens and timed these alterations over the past 35-55 million years. PMID- 11399079 TI - Holliday junction resolving enzymes of archaeal viruses SIRV1 and SIRV2. AB - In the final stages of genetic recombination, Holliday junction resolving enzymes transform the four-way DNA intermediate into two duplex DNA molecules by introducing pairs of staggered nicks flanking the junction. This fundamental process is apparently common to cells from all three domains of life. Two cellular resolving enzymes from extremely thermophilic representatives of both kingdoms of the domain Archaea, the euryarchaeon Pyrococcus furiosus and the crenarchaeon Sulfolobus solfataricus, have been described recently. Here we report for the first time the isolation, purification and characterization of Holliday junction cleaving enzymes (Hjc) from two archaeal viruses. Both viruses, SIRV1 and SIRV2, infect Sulfolobus islandicus. Their Hjcs both consist of 121 amino acid residues (aa) differing only by 18 aa. Both proteins bind selectively to synthetic Holliday-structure analogues with an apparent dissociation constant of 25 nM. In the presence of Mg(2+) the enzymes produce identical cleavage patterns near the junction. While S. islandicus shows optimal growth at about 80 degrees C, the nucleolytic activities of recombinant SIRV2 Hjc was highest between 45 degrees C and 70 degrees C. Based on their specificity for four-way DNA structures the enzymes may play a general role in genetic recombination, DNA repair and the resolution of replicative intermediates. PMID- 11399080 TI - The alternatively folded state of the antibody C(H)3 domain. AB - The C(H)3 domain of antibodies is characterized by two antiparallel beta-sheets forming a disulfide-linked sandwich-like structure. At acidic pH values and low ionic strength, C(H)3 becomes completely unfolded. The addition of salt transforms the acid-unfolded protein into an alternatively folded state exhibiting a characteristic secondary structure. The transition from native to alternatively folded C(H)3 is a fast reaction. Interestingly, this reaction involves the formation of a defined oligomer consisting of 12-14 subunits. Association is completely reversible and the native dimer is quantitatively reformed at neutral pH. This alternatively folded protein is remarkably stable against thermal and chemical denaturation and the unfolding transitions are highly cooperative. With a t(m) of 80 degrees C, the stability of the alternatively folded state is comparable to that of the native state of C(H)3. The defined oligomeric structure of C(H)3 at pH 2 seems to be a prerequisite for the cooperative unfolding transitions. PMID- 11399081 TI - Protein-dependent transition states for ribonucleoprotein assembly. AB - Native folding and splicing by the Saccharomyces cerevisiae mitochondrial bI5 group I intron RNA is facilitated by both the S. cerevisiae CBP2 and Neurospora crassa CYT-18 protein cofactors. Both protein-bI5 RNA complexes splice at similar rates, suggesting that the RNA active site structure is similar in both ribonucleoproteins. In contrast, the two proteins assemble with the bI5 RNA by distinct mechanisms and bind opposing, but partially overlapping, sides of the group I intron catalytic core. Assembly with CBP2 is limited by a slow, unimolecular RNA folding step characterized by a negligible activation enthalpy. We show that assembly with CYT-18 shows four distinctive features. (1) CYT-18 binds stably to the bI5 RNA at the diffusion controlled limit, but assembly to a catalytically active RNA structure is still limited by RNA folding, as visualized directly using time-resolved footprinting. (2) This mechanism of rapid stable protein binding followed by subsequent assembly steps has a distinctive kinetic signature: the apparent ratio of k(off) to k(on), determined in a partitioning experiment, differs from the equilibrium K(d) by a large factor. (3) Assembly with CYT-18 is characterized by a large activation enthalpy, consistent with a rate limiting conformational rearrangement. (4) Because assembly from the kinetically trapped state is faster at elevated temperature, we can identify conditions where CYT-18 accelerates (catalyzes) bI5 RNA folding relative to assembly with CBP2. PMID- 11399082 TI - Biochemical characterization of the structure-specific DNA-binding protein Cmb1 from Schizosaccharomyces pombe. AB - Cmb1, a novel HMG box protein from Schizosaccharomyces pombe, has been characterized biochemically using glutaraldehyde cross-linking, gel-filtration and analytical ultracentrifugation. It was identified as a monomeric, non spherical protein, with a tendency to aggregate in solution. Limited proteolysis with trypsin and chymotrypsin showed that the C-terminal HMG box was a compact, proteolytically stable domain and the N-terminal region of Cmb1 was relatively unstructured and more easily digested. As Cmb1 was previously identified as a potential mismatch-binding protein, the binding constants and stoichiometry for both homoduplex and heteroduplex DNA were determined using an IASys resonant mirror biosensor. Cmb1 indeed demonstrated a tighter association with mismatched DNA, especially with the C/Delta-mismatch. Expression constructs of Cmb1 were made to study the sections of the protein involved in DNA binding. Constructs with the N-terminal region absent revealed that the C-terminal HMG box was the primary DNA-binding region. The presence of the N-terminal region did, however, facilitate tighter binding to both homoduplex and heteroduplex DNA. The amino acid residues isoleucine 14 and leucine 39 were located as putative intercalating residues using structure guided homology modelling. The model templates were derived from two distinct HMG:DNA complexes: HMG-D bound to homoduplex DNA and HMG 1 bound to cisplatin DNA. Binding studies using the Cmb1 HMG box with point mutations in these residues showed that isoleucine 14 was important for the binding of Cmb1 to homoduplex DNA, but affected binding to mismatches to a lesser extent. In contrast, leucine 39 appeared to have a more significant function in binding to mismatched DNA. PMID- 11399083 TI - Folded-back solution structure of monomeric factor H of human complement by synchrotron X-ray and neutron scattering, analytical ultracentrifugation and constrained molecular modelling. AB - Factor H (FH) is a regulatory cofactor for the protease factor I in the breakdown of C3b in the complement system of immune defence, and binds to heparin and other polyanionic substrates. FH is composed of 20 short consensus/complement repeat (SCR) domains, for which the overall arrangement in solution is unknown. As previous studies had shown that FH can form monomeric or dimeric structures, X ray and neutron scattering was accordingly performed with FH in the concentration range between 0.7 and 14 mg ml(-1). The radius of gyration of FH was determined to be 11.1-11.3 nm by both methods, and the radii of gyration of the cross section were 4.4 nm and 1.7 nm. The distance distribution function P(r) showed that the overall length of FH was 38 nm. The neutron data showed that FH was monomeric with a molecular mass of 165,000(+/-17,000) Da. Analytical ultracentrifugation data confirmed this, where sedimentation equilibrium curve fits gave a mean molecular mass of 155,000(+/-3,000) Da. Sedimentation velocity experiments using the g*(s) derivative method showed that FH was monodisperse and had a sedimentation coefficient of 5.3(+/-0.1) S. In order to construct a full model of FH for scattering curve and sedimentation coefficient fits, homology models were constructed for 17 of the 20 SCR domains using knowledge of the NMR structures for FH SCR-5, SCR-15 and SCR-16, and vaccinia coat protein SCR-3 and SCR-4. Molecular dynamics simulations were used to generate a large conformational library for each of the 19 SCR-SCR linker peptides. Peptides from these libraries were combined with the 20 SCR structures in order to generate stereochemically complete models for the FH structure. Using an automated constrained fit procedure, the analysis of 16,752 possible FH models showed that only those models in which the 20 SCR domains were bent back upon themselves were able to account for the scattering and sedimentation data. The best-fit models showed that FH had an overall length of 38 nm and is flexible. This length is significantly less than a predicted length of 73 nm if the 20 SCR structures had been arranged in an extended arrangement. This outcome is attributed to several long linker sequences. These bent-back domain structures may correspond to conformational flexibility in FH and enable the multiple FH binding sites for C3 and heparin to come into close proximity. PMID- 11399084 TI - Crystal structure of 2-hydroxyl-6-oxo-6-phenylhexa-2,4-dienoic acid (HPDA) hydrolase (BphD enzyme) from the Rhodococcus sp. strain RHA1 of the PCB degradation pathway. AB - 2-Hydroxyl-6-oxo-6-phenylhexa-2,4-dienoic acid (HPDA) hydrolase (the BphD enzyme) hydrolyzes a ring-cleavage product of an aromatic compound generated in a biphenyl/polychlorinated biphenyl (PCB) degradation pathway of bacteria. The crystal structure of the BphD enzyme has been determined at 2.4 A resolution by the multiple isomorphous replacement method. The final refined model of the BphD enzyme yields an R-factor of 17.5 % at 2.4 A resolution with reasonable geometry. The BphD enzyme is an octameric enzyme with a 422 point-group symmetry. The subunit can be divided into core and lid domains. The active site of the enzyme is situated in the substrate-binding pocket, which is located between the two domains. The substrate-binding pocket can be divided into hydrophobic and hydrophilic regions. This feature of the pocket seems to be necessary for substrate binding, as the substrate is composed of hydrophilic and hydrophobic parts. The proposed orientation of the substrate seems to be consistent with the general catalytic mechanism of alpha/beta-hydrolases. PMID- 11399085 TI - Very high resolution structure of a trematode hemoglobin displaying a TyrB10 TyrE7 heme distal residue pair and high oxygen affinity. AB - Monomeric hemoglobin from the trematode Paramphistomum epiclitum displays very high oxygen affinity (P(50)<0.001 mm Hg) and an unusual heme distal site containing tyrosyl residues at the B10 and E7 positions. The crystal structure of aquo-met P. epiclitum hemoglobin, solved at 1.17 A resolution via multiwavelength anomalous dispersion techniques (R-factor=0.121), shows that the heme distal site pocket residue TyrB10 is engaged in hydrogen bonding to the iron-bound ligand. By contrast, residue TyrE7 is unexpectedly locked next to the CD globin region, in a conformation unsuitable for heme-bound ligand stabilisation. Such structural organization of the E7 distal residue differs strikingly from that observed in the nematode Ascaris suum hemoglobin (bearing TyrB10 and GlnE7 residues), which also displays very high oxygen affinity. The oxygenation and carbonylation parameters of wild-type P. epiclitum Hb as well as of single- and double-site mutants, with residue substitutions at positions B10, E7 and E11, have been determined and are discussed here in the light of the protein atomic resolution crystal structure. PMID- 11399086 TI - The origin of pH-dependent changes in m-values for the denaturant-induced unfolding of proteins. AB - Denaturant-induced unfolding is one of the most prevalent means of evaluating the structural stability of proteins and of determining the energetic consequences of mutations or changes in solution conditions. In spite of the widespread use of this approach, controversies and inconsistencies still persist with regard to the interpretation of the results of such studies. For example, most proteins show either a significant increase or a decrease (as much as 100 %) in the denaturant dependence of the free energy of unfolding (i.e. the m-value) under increasingly acidic conditions. The pH dependence of the m-value is given different interpretations depending on whether the m-values increase or decrease with decreasing pH. In cases where m-values decrease, the decrease is attributed to the presence of an intermediate that becomes transiently stabilized during the unfolding transition at low pH. Cases where m-values increase as pH is lowered are usually interpreted in terms of an increase in the amount of surface area exposed by the denatured state at low pH. We have developed a general thermodynamic model that accounts for both types of behavior in terms of an intermediate that is populated throughout the unfolding transition. The model provides a unified framework for explaining both types of observed behavior, and the validity of the model was tested through the analysis of the pH dependence of m-values of staphylococcal nuclease. According to the model, the observed increase in m-values with decreasing pH is consistent with the existence of an intermediate that is populated during urea and guanidine unfolding. The intermediate becomes less populated during the unfolding transition at lower pH values giving rise to the apparent increase in m-values. These results argue that the prevailing interpretation need not apply to all proteins. PMID- 11399087 TI - Snapshots of protein folding. A study on the multiple transition state pathway of cytochrome c(551) from Pseudomonas aeruginosa. AB - Cytochrome c(551) (cyt c(551)) from Pseudomonas aeruginosa is a small protein (82 residues) that folds via a three-state pathway with the accumulation in the microsecond time-range of a compact collapsed intermediate. The presence of a single His residue, at position 16, permits the study of the refolding at pH 7.0 in the absence of miscoordination events. Here, we report on folding kinetics in the millisecond time-range as a function of urea under different pH conditions. Analysis of this process (over-and-above proline cis-trans isomerization) at pH 7.0, suggests the existence of a multiple transition state pathway in which we postulate three transition states. Taking advantage of site-directed mutagenesis we propose that the first "unfolded-like" transition state (t(1)) originates from the electrostatic properties of the collapsed state, while the second transition state (t(2)) involves the interaction between the N and C-terminal helices and is stabilized by the salt bridge between Lys10 and Glu70 ( approximately 1 kcal mol( 1)). Our results suggest that, contrary to other cytochromes c, the roll-over effect observed for cyt c(551) at low denaturant concentration can be interpreted in terms of a broad energy barrier without population of any intermediates. The third and more "native-like" transition state (M) can be associated with the breaking/formation of the Fe(3+)-Met61 bond. This strong interaction is stabilized by the hydrogen bond between Trp56 and heme propionate 17 (HP-17) as suggested by the increase in the unfolding rate at high denaturant concentration of the Trp56Phe site-directed mutant. PMID- 11399088 TI - The activity of the murine DNA methyltransferase Dnmt1 is controlled by interaction of the catalytic domain with the N-terminal part of the enzyme leading to an allosteric activation of the enzyme after binding to methylated DNA. AB - The mammalian DNA methyltransferase Dnmt1 is responsible for the maintenance of the pattern of DNA methylation in vivo. It is a large multidomain enzyme comprising 1620 amino acid residues. We have purified and characterized individual domains of Dnmt1 (NLS-containing domain, NlsD, amino acid residues: 1 343; replication foci-directing domain, 350-609; Zn-binding domain (ZnD), 613 748; polybromo domain, 746-1110; and the catalytic domain (CatD), 1124-1620). CatD, ZnD and NlsD bind to DNA, demonstrating the existence of three independent DNA-binding sites in Dnmt1. CatD shows a preference for binding to hemimethylated CpG-sites; ZnD prefers methylated CpGs; and NlsD specifically binds to CpG-sites, but does not discriminate between unmethylated and methylated DNA. These results are not compatible with the suggestion that the target recognition domain of Dnmt1 resides in the N terminus of the enzyme. We show by protein-protein interaction assays that ZnD and CatD interact with each other. The isolated catalytic domain does not methylate DNA, neither alone nor in combination with other domains. Full-length Dnmt1 was purified from baculovirus-infected insect cells. Under the experimental conditions, Dnmt1 has a strong (50-fold) preference for hemimethylated DNA. Dnmt1 is stimulated to methylate unmodified CpG sites by the addition of fully methylated DNA. This effect is dependent on Zn, suggesting that binding of methylated DNA to ZnD triggers the allosteric activation of the catalytic center of Dnmt1. The allosteric activation model can explain kinetic data obtained by others. It suggests that Dnmt1 might be responsible for spreading of methylation, a process that is observed during aging and carcenogenesis but may be important for de novo methylation of DNA. PMID- 11399089 TI - Enzymatic properties of recombinant Dnmt3a DNA methyltransferase from mouse: the enzyme modifies DNA in a non-processive manner and also methylates non-CpG [correction of non-CpA] sites. AB - We present the first in vitro study investigating the catalytic properties of a mammalian de novo DNA methyltransferase. Dnmt3a from mouse was cloned and expressed in Escherichia coli. It was shown to be catalytically active in E. coli cells in vivo. The methylation activity of the purified protein was highest at pH 7.0 and 30 mM KCl. Our data show that recombinant Dnmt3a protein is indeed a de novo methyltransferase, as it catalyzes the transfer of methyl groups to unmethylated substrates with similar efficiency as to hemimethylated substrates. With oligonucleotide substrates, the catalytic activity of Dnmt3a is similar to that of Dnmt1: the K(m) values for the unmethylated and hemimethylated oligonucleotide substrates are 2.5 microM, and the k(cat) values are 0.05 h(-1) and 0.07 h(-1), respectively. The enzyme catalyzes the methylation of DNA in a distributive manner, suggesting that Dnmt3a and Dnmt1 may cooperate during de novo methylation of DNA. Further, we investigated the methylation activity of Dnmt3a at non-canonical sites. Even though the enzyme shows maximum activity at CpG sites, with oligonucleotide substrates, a high methylation activity was also found at CpA sites, which are modified only twofold slower than CpG sites. Therefore, the specificity of Dnmt3a is completely different from that of the maintenance methyltransferase Dnmt1, which shows a 40 to 50-fold preference for hemimethylated over unmethylated CpG sites and has almost no methylation activity at non-CpG sites. PMID- 11399090 TI - Structural assembly of the active site in an aldo-keto reductase by NADPH cofactor. AB - A 1.9 A resolution X-ray structure of the apo-form of Corynebacterium 2,5-diketo d-gluconic acid reductase A (2,5-DKGR A), a member of the aldo-keto reductase superfamily, has been determined by molecular replacement using the NADPH-bound form of the same enzyme as the search model. 2,5-DKGR A catalyzes the NADPH dependent stereo-specific reduction of 2,5-diketo-d-gluconate (2,5-DKG) to 2-keto l-gulonate, a precursor in the industrial production of vitamin C. An atomic resolution structure for the apo-form of the enzyme, in conjunction with our previously reported high-resolution X-ray structure for the holo-enzyme and holo/substrate model, allows a comparative analysis of structural changes that accompany cofactor binding. The results show that regions of the active site undergo coordinated conformational changes of up to 8 A. These conformational changes result in the organization and structural rearrangement of residues associated with substrate binding and catalysis. Thus, NADPH functions not only to provide a hydride ion for catalytic reduction, but is also a critical structural component for formation of a catalytically competent form of DKGR A. PMID- 11399091 TI - gyrB-225, a mutation of DNA gyrase that compensates for topoisomerase I deficiency: investigation of its low activity and quinolone hypersensitivity. AB - The B subunit of DNA gyrase (GyrB) consists of a 43 kDa N-terminal domain, containing the site of ATP binding and hydrolysis, and a 47 kDa C-terminal domain that is thought to play a role in interactions with GyrA and DNA. In cells containing a deletion of topA (the gene encoding DNA topoisomerase I) a compensatory mutation is found in gyrB. This mutation (gyrB-225) results in a two amino acid insertion in the N-terminal domain of GyrB. We found that cells containing this mutation are more sensitive than wild-type cells to quinolone drugs with respect to bacteriostatic and lethal action. We have characterised the mutant GyrB protein in vitro and found it to have reduced DNA supercoiling, relaxation, ATPase, and cleavage activities. The mutant enzyme is up to threefold more sensitive to quinolones than wild-type. The mutation also increases the affinity of GyrB for GyrA and DNA, while the affinity of quinolone for the enzyme DNA complex is unaffected. We propose that the loss in activity is due to misfolding of the GyrB-225 protein, providing an example in which misfolding of one protein, DNA gyrase, suppresses a deficiency of another, topoisomerase I. The increased quinolone sensitivity is proposed to be a consequence of an altered conformation of the protein that renders quinolones better able to disrupt, rather than generate, gyrase-drug-DNA complexes. PMID- 11399092 TI - Activation of stress-activated protein kinase in osteoarthritic cartilage: evidence for nitric oxide dependence. AB - OBJECTIVE: We have demonstrated in bovine chondrocytes that nitric oxide (NO) mediates IL1 dependent apoptosis under conditions of oxidant stress. This process is accompanied by activation of c-Jun NH2-terminal kinase (JNK; also called stress-activated protein kinase). In these studies we examined activation of JNK in explant cultures of human osteoarthritic cartilage obtained at joint replacement surgery and we characterized the role of peroxynitrite to act as an upstream trigger. DESIGN: A novel technique to isolate chondrocyte proteins (<10% of total cartilage protein) from cartilage specimens was developed. It was used to analyse JNK activation by a western blot technique. To examine the hypothesis that chondrocyte JNK activation is a result of increased peroxynitrite, in vitro experiments were performed in which cultured chondrocytes were incubated with this oxidant. RESULTS: Activated JNK was detected in the cytoplasm of osteoarthritis (OA) affected chondrocytes but not in that of controls. In vitro, chondrocytes produce NO and superoxide anion. IL-1 (48 h), which induces nitric oxide synthase, resulted in an activation of JNK; this effect was reversed by N monomethylarginine (NMA). TNFalpha treated chondrocytes at 48 h produce superoxide anion (EPR method). Exposure of cells to peroxynitrite led to an accumulation of intracellular oxidants, in association with JNK activation and cell death by apoptosis. CONCLUSION: We suggest that JNK activation is among the IL-1 elicited responses that injure articular chondrocytes and this activation of JNK is dependent on intracellular oxidant formation (including NO peroxynitrite). In addition, the extraction technique here described is a novel method that permits the quantitation and study of proteins such as JNK involved in the signaling pathways of chondrocytes within osteoarthritic cartilage. PMID- 11399093 TI - Tissue inhibitor of metalloprotease-1 (TIMP-1) serum level may predict progression of hip osteoarthritis. AB - OBJECTIVE: To determine the predictive value of serum levels of TIMP-1 and hyaluronic acid in a 1 year prospective study in hip osteoarthritis (OA). METHODS: Twenty-nine patients with OA of the hip were enrolled in a 1-year prospective study (median follow-up, 13 months). Biochemical analysis was used to assess TIMP-1 and hyaluronic acid at entry and at the end of the study. Radiographic evaluation with an assisted computed program was performed to calculate progression of joint space narrowing. Statistical tests served to determine correlations between observed serum levels and radiograph joint space narrowing. RESULTS: Among the 29 patients, 10 showed joint space narrowing greater than 0.6 mm per year. The initial concentration of TIMP-1 as well as delta value of variation in serum levels of TIMP-1 (difference between TIMP-1 concentration at entry and at the end) correlated with the progression of joint space narrowing. A cut-off value of 600 ng/ml of TIMP-1 allowed the patients who progressed slowly from those who progressed more rapidly. Hyaluronic acid serum level was not predictive of evolution. CONCLUSION: TIMP-1 serum level may serve to predict the evolution of patients with hip OA. PMID- 11399094 TI - Type II collagen degradation in articular cartilage fibrillation after anterior cruciate ligament transection in rats. AB - OBJECTIVE: To investigate the kinetics of early cartilage changes in mechanically induced osteoarthritis (OA) and the association of these changes with damage to the type II collagen network. METHODS: Experimental OA was induced by anterior cruciate ligament transsection in the rat knee joint (ACLT-OA). Animals were sacrificed after 2, 7, 14, 28 and 70 days. Knee joints were evaluated using routine histology and immunohistochemistry for denatured (unwound) type II collagen to detect collagen damage. An antibody recognizing the collagenase cleavage site in type II collagen was used to study the role of collagenase in this process. RESULTS: The first changes of the articular cartilage after anterior cruciate ligament transection occurred in the superficial zone. These changes included loss of superficial chondrocytes, swelling of the remaining chondrocytes and superficial fibrillation. The swelling of the chondrocytes did not result from a change towards the hypertrophic phenotype, since these cells did not stain for type X collagen. A marked increase in denatured type II collagen staining was present in the fibrillated areas. Staining of the collagenase cleavage site showed the same distribution as denatured collagen but was clearly less intense. Collagen damage could never be detected before fibrillation occurred and was not present in non-fibrillated areas. CONCLUSIONS: These results indicate that in this model cartilage degeneration starts at the articular surface and that this degeneration is associated with a localized expression of type II collagen degradation products. PMID- 11399095 TI - Long-term outcome of meniscectomy: symptoms, function, and performance tests in patients with or without radiographic osteoarthritis compared to matched controls. AB - OBJECTIVE: To describe the long-term influence of meniscectomy on pain, functional limitations, and muscular performance. To assess the effects of radiographic osteoarthritis (OA), gender and age on these outcomes in patients with meniscectomy. DESIGN: 159 subjects (35 women), mean age 53 years, were examined 19 (17-22) years after open meniscectomy. Self-reported symptoms and function were assessed, performance tests were carried out and radiographs were taken. Sixty-eight age- and gender-matched controls were examined likewise. The data was analysed in two steps. First, subjects with meniscectomy were compared to the controls, and subgroup analyses were carried out with regard to radiographic OA, gender and age. Second, similar comparisons were carried out within the meniscectomized group. RESULTS: Meniscectomized subjects reported significantly (P< 0.001) more symptoms and functional limitations than did controls. This was also true when operated subjects without OA were compared to controls without OA. Within the meniscectomized group, severe radiographic OA (joint space narrowing grade 2 or more) and female gender, but not older age, was associated with more symptoms and functional limitations. Meniscectomy was associated with worse muscular performance. Female gender and older age were associated with worse muscular performance in the study group. CONCLUSIONS: Meniscectomy is associated with long-term symptoms and functional limitations, especially in women. Patients who had developed severe radiographic OA experienced more symptoms and functional limitations. Age did not influence self reported outcomes, however older age was associated with worse muscular performance. PMID- 11399096 TI - Differential regulation of gelatinases by transforming growth factor beta-1 in normal equine chondrocytes. AB - OBJECTIVE: Cartilage destruction in osteoarthritis (OA) is associated with increased levels of several matrix metalloproteinases (MMPs), including the gelatinases MMP-2 and MMP-9. While increases in some MMPs may be destructive, up regulation of others may result from increases in normal tissue turnover. The production of MMP-2 and MMP-9 by the anabolic transforming growth factor beta-1 (TGF-beta1) in normal equine chondrocytes was investigated. DESIGN: Equine chondrocytes from clinically normal femoropatellar joints were maintained in alginate beads. After serum deprivation, cells were exposed to TGF-beta1 at several concentrations for varying times. Activities of MMP-2 and MMP-9 were estimated by gelatin zymography, while mRNA for MMP-2, MMP-9 and collagen type II were detected using reverse transcription-polymerase chain reaction. RESULTS: Stimulation with TGF-beta1 differentially regulated MMP-2 and MMP-9, with strong up-regulation of both MMP-9 mRNA and enzyme. Increases in MMP-9 enzyme were dose dependent (0-49 h). There is some evidence suggesting a slight reduction in MMP-2 release following stimulation. Collagen type II mRNA was transiently increased following stimulation. CONCLUSIONS: The different effects of TGF-beta1 on MMP-2 and MMP-9 production by normal chondrocytes suggests different roles for these enzymes. The increases in both MMP-9 and collagen type II mRNA following stimulation may suggest a role for MMP-9 in tissue maintenance. Therefore, increased MMP-9 may be secondary to, as distinct from a cause of, cartilage damage. PMID- 11399097 TI - Nimesulide reduces interleukin-1beta-induced cyclooxygenase-2 gene expression in human synovial fibroblasts. AB - OBJECTIVE: To characterize the effects of nimesulide (NIM) on basal and induced cyclo-oxygenase-2 (COX-2) gene expression in human synovial fibroblasts (HSF) and to define the intracellular mechanisms that mediate the changes in COX-2 expression and synthesis in response to the drug. DESIGN: HSF were incubated with NIM and NS-398 (0, 0.03, 0.3, 3 microg/ml) in the absence or presence of the COX 2 inducers interleukin-1beta (IL-1beta) or endotoxin (LPS). Treated cells were analysed for COX-2 mRNA and protein by Northern and Western blotting analysis, respectively. Putative transcriptional, post-transcriptional, and signaling effects of NIM on basal and induced-COX-2 expression were investigated by human COX-2 promoter studies, calcium studies, reactive oxygen species (ROS) evaluations, electrophoretic mobility shift analysis (EMSA) and half-life studies of COX-2 mRNA. RESULTS: NIM inhibited IL-1beta-induced COX-2 expression and protein at sub and therapeutic concentrations (0.03-0.3 microg/ml) while the non specific NSAID, naproxen, did not. Both drugs suppressed PGE2 release by about 95%. NIM had no effect on (1) IL-1beta-induced increases in NF-kappaB or c/EBP signaling, or (2) human COX-2 promoter activity. Stability of induced COX-2 mRNA was unaffected by NIM treatments. Pre-treatment of cells with O(2)radical scavengers (e.g. PDTC) or with Ca(++)channel blockers (e.g. verapamil) had a modest effect on IL-1beta-induced COX-2 expression. NIM blocked ionomycin+thapsigargin and H(2)O(2)-induced increases in COX-2 protein synthesis. CONCLUSION: NIM inhibits cytokine-induced COX-2 expression and protein at sub and therapeutic concentrations. At least part of this activity may be the result of NIM inhibition of calcium and/or free radical generation induced by cytokines. PMID- 11399098 TI - Biological freezing of human articular chondrocytes. AB - AIM: To preserve viable, metabolically active chondrocytes cultured in alginate beads at -196 degrees C for further use in in vitro and in vivo studies. METHODS: Human articular chondrocytes were isolated from femoral condyles within 24 h post mortem. To optimize the biological freezing procedure, the chondrocytes were control-rate frozen in different concentrations of dimethyl sulfoxide (DMSO) in Dulbecco's MEM supplemented with 10% FCS before being thawed and the cell viability was determined by Trypan Blue exclusion test. To investigate the effect of control-rate freezing on chondrocyte metabolism, control-rate frozen chondrocytes in 5% DMSO were thawed and cultured in gelled agarose for 2 weeks. Non-frozen chondrocytes cultured in agarose served as controls. Furthermore, human articular chondrocytes were cultured in 2% alginate beads for 2 weeks after which the beads were incubated with 5% DMSO for 0 h, 2.5 h, 5 h and 10 h and frozen at -196 degrees C. Non-frozen alginate beads containing chondrocytes and incubated with 5% DMSO served as a control. After 2 weeks in culture, chondrocytes in agarose or in alginate were sulfated with 10 microCi(35)SO(4)/ml for 48 h. The total production of aggrecans, and the aggrecan subtypes, were subsequently determined. RESULTS: Five percent DMSO in the culture medium was the optimal condition to control-rate freeze and recover viable and functional isolated chondrocytes. Total aggrecan synthesis of control-rate frozen chondrocytes cultured in gelled agarose was not significantly reduced when compared with control cells. The proportion of aggrecan in the aggregate form of control-rate frozen chondrocytes kept in agarose remained unaltered. Chondrocytes, control-rate frozen in the alginate matrix, showed a 0-30% decrease in total aggrecan synthesis rates in culture when compared with the non-frozen chondrocytes. The optimal pre-incubation time of the alginate beads with 5% DMSO was 5 h, without any change in aggrecan synthesis rates when compared with the control situation. Shorter pre-incubation times resulted in an insufficient diffusion of DMSO into the beads and in cell death. There was no difference in the synthesis of the different aggrecan subtypes between frozen and non-frozen chondrocytes in alginate. CONCLUSION: Human articular chondrocytes can be stored at -196 degrees C for 24 h without important decreases in their aggrecan synthesis rates when control-rate frozen as a cell suspension in 5% DMSO. Proportions of the aggrecan subtypes (monomers, aggregates) synthesized by chondrocytes cultured in agarose remained unchanged. The control-rate freezing procedure in the alginate beads pre-incubated with 5% DMSO for 5 h produced no decrease in total aggrecan synthesis rates and no change in the synthesized aggrecan subtypes. Further experiments have to confirm the suitability of this freezing method for long-term storage of chondrocytes allowing us to set up a 'chondrocyte' bank for further use in in vitro and in vivo manipulations. PMID- 11399099 TI - Effects of high molecular weight hyaluronan on the distribution and movement of proteoglycan around chondrocytes cultured in alginate beads. AB - OBJECTIVE: To evaluate the effects of high molecular weight hyaluronan (HA) on the distribution and movement of proteoglycan (PG) formed around rabbit chondrocytes cultured in alginate beads. DESIGN: Rooster comb-derived HA (MW 8x10(5) Da) was co-polymerized in alginate gel to study the direct effects of extrinsic HA on chondrocytes. PG metabolism of rabbit chondrocytes cultured in alginate beads was examined by measuring the incorporation of [(35)S]sulfate into glycosaminoglycan in two distinct regions, the cells with their cell-associated matrix (CM) and the further-removed matrix (FRM). Immunohistochemical analysis was performed using monoclonal antibodies against chondroitin sulfate and keratan sulfate. Autoradiography using degenerated cartilage tissue from the rabbit osteoarthritis (OA) model was performed to discover the effect of HA on the distribution of newly-synthesized PG in the cartilage tissue. RESULTS: The incorporation of [(35)S]sulfate into newly-synthesized PG in the cells with CM decreased with the addition of 0.125-1.0 mg/ml HA, while the incorporation in the FRM increased. These effects of HA on the distribution of newly-synthesized PG were the same either in chondrocytes with CM or chondrocytes without CM. Immunohistochemical analysis showed that staining of PG in the CM was decreased and staining in the FRM was increased in the HA treated group compared to the control group. Autoradiography using degenerated cartilage tissue from the rabbit OA model indicated that [(35)S]-labeled macromolecules showed a more diffuse distribution in the HA treated group compared with the control group. CONCLUSION: These results indicate that extrinsic HA could affect the movement of newly synthesized PG from the CM to the FRM in both alginate beads and cartilage tissue. PMID- 11399100 TI - Modulation of bovine articular chondrocyte gene expression in vitro by oxygen tension. AB - OBJECTIVE: Adult articular cartilage is a physiologically hypoxic tissue with a proposed gradient of oxygen tension ranging from <10% oxygen at the cartilage surface to <1% in the deepest layers. This gradient may be disturbed during diseases of the joint, for example in rheumatoid arthritis when synovial fluid pO(2)falls. We investigated whether changes in oxygen tension modulate gene expression in articular chondrocytes. DESIGN: Bovine articular chondrocytes were cultured in alginate beads in medium maintained at <0.1, 5, 10 or 20% oxygen. A modified RNA arbitrarily primed polymerase chain reaction (RAP-PCR) technique was used to identify several genes whose mRNA abundance in articular chondrocytes was dependent upon oxygen tension. Northern hybridization slot blots were used to quantify changes in mRNA level relative to a housekeeping gene, beta-actin. RESULTS: Genes found by RAP-PCR to undergo up-regulation in hypoxia included TIMP 1 and integrin-linked kinase. Collagen V mRNA levels were down-regulated in hypoxic chondrocytes. This led us to examine mRNA levels for various cytokines, matrix structural molecules and beta1 integrin. Interleukin 1beta, transforming growth factor beta and connective tissue growth factor were all up-regulated by low oxygen tensions, as was beta1 integrin. Collagen II (COL2A1) was down regulated by hypoxia but aggrecan mRNA levels remained unchanged. The mRNA levels for GAPDH, the archetypal hypoxia responsive gene, were not modulated in articular chondrocytes by changes in oxygen tension. CONCLUSIONS: Oxygen tension modulates the abundance of mRNAs encoding structural molecules, several cytokines, beta1 integrin and integrin-linked kinase in articular chondrocytes. This may be important during disease progression. Chondrocytes are unusual in their response to hypoxia, presumably because they exist physiologically in a low oxygen environment. PMID- 11399101 TI - Quantitation of chondroitin-sulfates, disaccharides and hyaluronan in normal, early and advanced osteoarthritic sheep temporomandibular joints. AB - OBJECTIVE: To determine the relationship between synovial fluid, chondroitin sulfate disaccharide and hyaluronic acid to differing degrees of experimental temporomandibular joint (TMJ) osteoarthritis (OA). DESIGN: Twenty-four merino sheep were divided into three groups and had different TMJ surgical procedures to produce OA. Group I; control (six sheep), Group II; disc perforation (nine sheep) and Group III; disc perforation and articular damage (nine sheep). Synovial fluid was collected initially and at sacrifice at 3 months. Chondroitin 4-sulfate, chondroitin 6-sulfate and hyaluronic acid were measured and correlated to the OA histologic score. RESULTS: The chondroitin-sulfate levels were significantly increased (Group I to Group II P< 0.001; Group I to Group III P< 0.001), the hyaluronic acid levels decreased (Group I to Group II P< 0.01; Group I to Group III P< 0.01) with the increasing OA score. CONCLUSION: Chondroitin-sulfate and hyaluronic acid show a correlation with surgically created TMJ osteoarthritis in sheep model. PMID- 11399102 TI - Morphological analysis of articular cartilage biopsies from a randomized, clinical study comparing the effects of 500-730 kDa sodium hyaluronate (Hyalgan) and methylprednisolone acetate on primary osteoarthritis of the knee. AB - OBJECTIVE: Histomorphometric study on cartilage samples taken from osteoarthritic human knees before and 6 months after intraarticular injections of a specific fraction (500-730 kDa) of hyaluronan. The results obtained with hyaluronan were compared with the results of methylprednisolone acetate treatment. METHODS: Twenty-four subjects with primary osteoarthritis (OA) of the knee were considered. Eleven patients were treated with Hyaluronan (Hyalgan), 20 mg/2 ml once a week for 5 weeks) and 13 with methylprednisolone (Depo-Medrol, 40 mg/1 ml once a week for 3 weeks). At the time of baseline and after 6 months from the start of treatment, biopsies of cartilage were taken and processed for electron microscopy. Articular surface morphology, territorial matrix, chondrocyte number and ultrastructure were characterized by a set of morphometric parameters. Samples from 19 informed patients showing no arthroscopic sign of OA were also used for comparison. RESULTS: Six months after hyaluronan treatment a significant reconstitution of the superficial layer were observed together with an improvement in chondrocyte density and territorial matrix appearance. Furthermore, chondrocytes appeared significantly improved in their metabolism, as indicated by the increased extension of the synthetic structures and mitochondria with respect to the organelles having catabolic or storage functions. Hyaluronan treatment produced results that were significantly superior to those delivered with Methylprednisolone in almost all the morphometric estimators. CONCLUSIONS: These results cannot be explained simply by temporary restoration of the synovial fluid viscoelasticity, and provide further evidence that the specific fraction of hyaluronan used in this study is a useful tool in OA treatment, with a potential structure-modifying activity. PMID- 11399103 TI - Growth factor expression in cartilage wound healing: temporal and spatial immunolocalization in a rabbit auricular cartilage wound model. AB - OBJECTIVE: The ability of cartilage to regenerate following injury is limited, potentially leading to osteoarthritis. Integrative cartilage repair, necessary for durable restoration of cartilage lesions, can be regarded as a wound healing process. Little is known about the effects of growth factors regulating acute cartilage wound healing in vivo. In this study the temporal expression patterns of growth factors and proteoglycan content in cartilage wound edges in vivo were studied. DESIGN: Cartilage wounds were created in rabbit ear cartilage using a 6 mm biopsy punch. Specimens were subsequently harvested 1, 3, 7, 14 and 28 days after surgery. Paraffin sections were thionin stained to visualize proteoglycan loss and replacement. Immunohistochemical staining of TGFbeta1, TGFbeta3, IGF-1, IGF-II and FGF-2 was used to define growth factor expression at the cartilage wound sites. RESULTS: Almost no effect of cartilage wounding was observed one day after surgery. A decrease of proteoglycan content, with a maximal loss at day 7, and a subsequent restoration was observed at the wound edges. Growth factor expression increased simultaneously. Maximal immunostaining for IGF1, IGFII, FGF2 and TGF-beta3 was observed at day 7, followed by a gradual decrease. Increased expression of TGFbeta1 lasted from day 3 until day 14. CONCLUSION: We have demonstrated the ability of chondrocytes to increase growth factor expression and to restore the rapid decrease in proteoglycan content in the initial phase following acute wounding. A temporal increase in intracellular growth factor expression suggests an autocrine and/or paracrine metabolic stimulation, which can be regarded a sign of chondrocytes repair capacity. PMID- 11399104 TI - Sodium hyaluronate inhibits interleukin-1-evoked reactive oxygen species of bovine articular chondrocytes. PMID- 11399105 TI - Gradual micronutrient accumulation and depletion in Alzheimer's disease. AB - Cadmium is a carcinogen that accumulates relentlessly with age, reaching high levels in the liver and kidneys. It is known to hyperactivate the Kupffer cells (hepatic macrophages). On the other hand, the risk of developing Alzheimer's disease increases considerably with age and it involves neuronal damage by hyperactive microglia (brain macrophages). Moreover, many of the metals that accumulate in the liver and kidneys, also accumulate in the brain (Fe, Cu, Zn, Mn, etc.). Therefore, it is possible that Cd also hyperactivates the microglia, playing a role in Alzheimer's disease (AD).Fe also accumulates in the brain as we age and catalyzes super oxide (O2-) formation, which reacts with nitric oxide (NO) to form the very harmful peroxynitrite (ONOO-). ONOO- causes considerable damage that exacerbates the damage caused by the hyperactive microglia, accelerating the progress of AD. Moreover, as we age we become less efficient at absorbing and retaining Cu, Zn and Mg. Since Cu and Zn are necessary for the synthesis of copper-zinc superoxide dismutase (CuZnSOD), which disables the noxious O2-, the deficiencies cause considerable damage as we age. Similarly, Mg is a cofactor for CuZnSOD and is necessary for NO to leave the cell and perform its vasodilating job. Unfortunately, a Mg deficiency traps the NO in the cell, where it reacts with O2-, forming the harmful ONOO-. Furthermore, Se and vitamins B6 and D are required for Mg absorption and vitamin E is required to minimize the oxidative damage. PMID- 11399106 TI - Biological processes, quantum mechanics and electromagnetic fields: the possibility of device-encapsulated human intention in medical therapies. AB - The general hypothesis that quantum mechanics (QM) and thermodynamic concepts relate to biological systems is discussed and applied to the biological influence of: (1) electromagnetic fields (EMFs); and (2) EMFs that have been exposed to human intention. We illustrate our hypothesis with experiments involving four simultaneous treatments: exposure to ambient EMFs in the laboratory environment (C), exposure in a Faraday cage (F) and exposure in a Faraday cage with either: (i) an electronic device (IIED) which had been exposed to a specific human intention (d,j); or (ii) a non-exposed, physically identical, device (d,o). Experimental systems were fitness and energy metabolism in Drosophila melanogaster, in vitro enzyme activity and molecular concentration variability over time. Results indicated that shielding from ambient EMFs via a Faraday cage (F) made a significant difference relative to the unshielded control (C). Further, (d,o) had a significant lowering effect in the shielded environment. Finally, there was a strong 'intention' effect with the IIED (d,j) producing significant and positive effects in comparison to (d,o) in each experimental system. PMID- 11399107 TI - A biochemical theory to explain the cause of bovine spongiform encephalopathy and other encephalopathies. AB - The purpose of this study is to present a hypothesis to explain the aetiology of bovine spongiform encephalopathy (BSE) which is more credible than any at present available, and to increase its credibility by varying the hypothesis to supply explanations for Alzheimer's disease, Parkinson's disease and certain other conditions. The method used has been to utilize material from biochemical textbooks and similar sources. It has been concluded that BSE is caused by the failure to synthesize sufficient cyclic guanosine monophosphate (cGMP), with the result that neurons die because they are no longer able to prevent the entry of toxic quantities of calcium ions into their cytoplasm. Several causes for the failure to synthesize sufficient cGMP have been identified; these involve selenium and folate deficiencies, and problems with the availability of nicotinamide adenosine dinucleotide (NAD). It is proposed that BSE is initiated by a combination of selenium deficiency and the destruction of NAD by a bacterial toxin of the same type as causes cholera, that folate deficiency is the predominant cause of Alzheimer's disease, and that the failure to synthesize sufficient tetrahydrobiopterin and cGMP from guanosine triphosphate results in Parkinson's disease. PMID- 11399108 TI - Increased superoxide dismutase and Down's syndrome. AB - The enzyme superoxide dismutase (SOD) is a constitutive enzyme coded by a gene located in Chromosome 21 (21q22.1). Thus, the tissues from patients with trisomy 21 contain 50% more SOD activity. It is often suggested that the increased SOD content in the cells of Down's syndrome patients is responsible for many of the neurological symptoms of this disease. This hypothesis is not supported by most of the available data. In this paper we discuss why the increased SOD activity should not influence neuronal function and propose, instead, that the main problem may be an overexpression of other genes also located in chromosome 21 such as the beta amyloid precursor, as well as oncogenes and other Down's syndrome-related genes. We also propose that the increased SOD may be, instead, responsible for the increased incidence of Down's syndrome in children of older women. The augmented antioxidant protection resulting from an extra copy of chromosome 21 may, with time, selectively protect human oocytes from apoptosis, increasing their proportion with age, explaining the higher incidence of this disease. PMID- 11399109 TI - Immediate and longterm effects of immune stimulation: hypothesis linking the immune response to subsequent physical and psychological wellbeing. AB - We hypothesize that antigenic stimuli in susceptible persons during key developmental life stages alters neuroendocrine-immune organization and leads to the development of aberrant immune and neuroendocrine responses to subsequent environmental stressors, with longterm physical and psychological consequences. The release of interleukin-1beta (IL-1beta) and other proinflammatory cytokines associated with the immune response during times when individuals are most vulnerable to the effects of environmental influences activates the hypothalamic pituitary-adrenal (HPA) axis and leads to maladaptive responses to subsequent stressors. The primed HPA axis is reactivated by proinflammatory cytokines, resulting in the secretion of corticotropin-releasing hormone (CRH) and cortisol, followed by physical and psychological effects that feedback on the HPA axis to produce an array of outcomes affecting general wellbeing. Through the release of histamine and other mediators and their effects on the mast cell-leukocyte cytokine cascade, immune stimuli in susceptible persons increase allergic inflammation and magnify stressors' effects through the release of HPA-axis activating cytokines, such as IL-1beta, that drive the axis and reinforce the physiological and behavioral effects. Thus, specific proinflammatory cytokines and allergic reactions initiate, promote, and maintain immune-stimulus-associated HPA axis activity, and with CRH and cortisol, participate in a positive feedback loop, resulting in aberrant, maladaptive responses to physical or psychological stressors, with outcomes such as depression, hyperalgesia, and pain-related behavior. PMID- 11399110 TI - Renal regulation of plasma total antioxidant capacity. AB - Hyperuricemia has been labeled both a risk factor and marker for cardiovascular pathology in addition to being associated with gout and kidney disease. Uric acid in vitro acts as a potent antioxidant capable of scavenging hydroxy radicals and peroxynitrite and reacting with nitric oxide. Some clinical studies have provided evidence that, in vivo, uric acid is oxidized under conditions associated with high oxidant stress and may spare other antioxidants such as ascorbic acid. The plasma level of uric acid is controlled by the rates of production and excretion or degradation of uric acid. Under most circumstances, it is the renal clearance of uric acid which primarily determines the plasma concentration. Many factors of exogenous and endogenous origin can influence renal tubular absorption and secretion of uric acid. We suggest that renal urate clearance is not haphazard but regulated by an unknown signal that is issued in response to the level of oxidative stress. Since much cardiovascular pathology is now believed to have an inflammatory component and is associated with enhanced production of free radicals, the accompanying hyperuricemia may be viewed as a compensatory response of potential benefit. PMID- 11399111 TI - Mechanisms for telomerase gene control in aging cells and tumorigenesis. AB - Although telomerase, which maintains the ends of chromosomes, is down-regulated as cells differentiate leading to attrition of chromosomal termini and ultimate replicative senescence, it is up-regulated in most cancer cells which show no net loss of average telomere length. The mRNA level of the catalytic component of telomerase, hTERT, is the major determinant of telomerase activity but little is known about control of hTERT transcription. We propose mechanisms whereby cytosine methylation may alter the binding of activators such as c-Myc or repressors such as WT1 which interact with the hTERT gene regulatory region to modulate telomerase activity in aging cells and tumorigenesis. Mechanisms are also proposed for control of hTERT expression through changes in the collective binding of its transcription factors in aging and tumorigenic cells. Elucidation of telomerase regulation should facilitate advances in understanding age-related diseases such as cancer and in potential therapeutic modalities. PMID- 11399112 TI - Are cannabinoid receptor knockout mice animal models for schizophrenia? AB - Schizophrenia is a devastating psychiatric disorder with a high prevalence worldwide. There is therefore a need for animal models allowing the development of new therapeutic interventions and reliable diagnostic tests. In the temporal domain, cannabinoid receptor gene (CB1) knockout mice exhibit behavioural alterations, which parallel symptoms in schizophrenia, cannabis intoxication and dopamine D2 activation. While a specific nucleotide homology between CB1 and D2 accounts for the pathophysiology, pre-inserted spirochaetal DNA on the polyadenylation signal of CB1 reveals the aetiology of schizophrenia. If, in analogy to thalassaemia, mutations occur within this 3' regulatory domain, the genetic expression of CB1 is disrupted and sequential information lost in time. CB1, previously unrecognized as a candidate gene, thus unifies the different aspects of schizophrenic psychosis: cannabis-induced model psychosis, disrupted information processing, spatio-temporal distortions and other psychotic symptoms, disturbed neuronal migration, schizophrenic brain disorder, familial transmission, and prenatal infection by Borrelia burgdorferi. PMID- 11399113 TI - Crosstalk and considerations in endocrine disruptor research. AB - The presence of endocrine disrupting chemicals in the environment has prompted action on several fronts to assess the potential health risks of these compounds. To fully understand the mechanisms behind the observed endocrine disruption, crosstalk and other factors should be considered. In this article we will discuss how crosstalk modulates estrogen action in several common assays and how this and other considerations appear to have been overlooked. In addition, a paradigm shift from theoretical linear response pathways to interaction maps should aid in the understanding and analysis of endocrine disruption. PMID- 11399114 TI - Suppression of immune surveillance in melanoma. AB - Numerous reports of IL-10 cytokine secretion by tumor infiltrating cells indicate there is suppression of immune surveillance within the milieu of many melanomas. In this paper we have outlined the suppressor system that best fits the published data. The regulatory system is composed of CD4+ T-lymphocytes which have been activated and programmed to secrete Th2 cytokines. Initially these cells do not secrete cytokines, but subsequently they enter an IL-10 secretory phase as a result of T-T cell interaction. After activation, Th1 programmed T-cells express MHC class II molecules and B7 second signals. When these Th1 T-cells express MHC II molecules containing 'self' polypeptides coupled with faulty B7-H1 second signals they are subject to inactivation by Th2 T-cells. If this system can be inactivated, immunotherapy of melanoma will be more successful. If an antigen can be discovered that stimulates sensitized Th2 T-cells without stimulating Th1 T cells, this antigen, followed with cyclophosphamide, can be used to destroy the Th2 T-cells in the course of both active and passive immunotherapy. Solubilized MHC II molecules with appropriate 'self' polypeptides should qualify as such an antigen. We postulate such an antigen can be prepared using poliovirus 1 (Sabin) to lyse melanoma tissue cultures. PMID- 11399115 TI - Breathing pauses during sleep: can a non-invasive ENT examination help identify subjects at risk in epidemiological settings? AB - In patients with obstructive sleep apnoea (OSA) anatomic and functional upper airway abnormalities are frequent and severe. Invasive methods are used to identify and quantitate the obstruction, to precisely locate its site, etc. as part of pre-treatment or of preoperative evaluations.These methods (lateral skull radiographs, computerized tomography, MRI, fibroscopies, etc) are too expensive and too invasive to be utilized in field surveys. To the classical sleep questionnaires and anthropometric measurements, some simple nose-throat examinations, easily accepted by the volunteers in a population study, could add useful information for the identification of the subjects at risk for sleep disordered breathing. The present paper is a review of these examinations and of their utility. PMID- 11399116 TI - Hypophosphataemia: cause of the disturbed metabolism in the metabolic syndrome. AB - The clustering of risk factors for cardiovascular disease (CVD) and non-insulin dependent diabetes mellitus (NIDDM) in obese patients may be attributable to a disturbed metabolism caused by hypophosphataemia. A low serum phosphate (S-P) level may be a limiting factor for glucose metabolism and may account for hyperglycaemia, with an increased risk of NIDDM and hypertension and consequent increased risk of stroke. Low S-HDL levels, known to exist in the metabolic syndrome, as well as high serum triglycerides may also have been the results of phosphate depletion. The hypothesis presents a new serious disturbance which accounts for the dyslipidaemia, hyperglycaemia and the hypertension in metabolic syndrome. The proposed causal relationship between low S-P and the clustering of risk factors is based on results from a cross-sectional study of obese patients, where low S-P was associated with high body mass index (BMI), high blood glucose (B-glu), high systolic blood pressure (SBP), high diastolic blood pressure (DBP), but low serum high density lipoprotein (S-HDL) and serum magnesium (S-Mg) levels. Knowledge from experimental and clinical studies on hypophosphataemia and/or phosphate depletion are referred to when discussing the hypothesis. PMID- 11399117 TI - Effects of antisocial or social attitudes on neurobiological functions. AB - Antisocial personality disorder is characterized by triats like a failure to conform to social norms, impulsivity, hostility, irritability and aggressiveness, recklessness, irresponsibility, criminality, a lack of guilt or remorse, and deceitfulness. A large number of investigations revealed that there is evidence for a neurobiological underpinning of antisocial behavior. The precise nature and causal aspects of the relationship between neurobiological abnormalities and antisocial behavior is still unknown. There is, however, some evidence that some antisocial patients are able to influence their neurobiological functions, which are related to sensation seeking, aggressiveness, impulsivity, criminality and a lack of fear (associated with an inability to learn from experiences). The author hypothesizes that a reduction of antisocial behavior/attitude, may consequently result in normalization of neurobiological functions, which are linked to specific attitudes and behavioral dimensions. PMID- 11399118 TI - Neuronal hypersynchronization, creativity and endogenous psychoses. AB - I have investigated a neuronal hypersynchronism, currently included under the general subject of epilepsy, and termed interictal activity. I suggest that it is a physiological activity of the mammalian brain and propose it be termed Hyperia. After a thorough study of the extraordinary psychic manifestations of this neuronal hypersynchronism shown by mystics and artists, I have reviewed several scientific publications bearing on my hypothesis. I conclude by elaborating on a variety of cerebral hypersynchronous functions whose cause I consider to be physiological. Such behaviour is a common basis for extraordinary psychic manifestations found not only in mystics and artists, but also in patients suffering from endogenous psychoses, especially Bipolar Disorder. PMID- 11399119 TI - Eicosanoids: the molecules of evolution. AB - The molecules of evolution constitute a response system linking the environment to the internal machinery of cells and are key factors in evolutionary disorders. They orchestrate such mechanisms of evolution as replication and reproduction, variation, inheritance, natural selection, heterochrony, neoteny, aging, recapitulation, metamorphosis, homology, paradox, and extinction. Eicosanoids fulfill these criteria. PMID- 11399120 TI - A possible role of decreased oxidative resistance of low-density lipoproteins in the early formation of carotid atherosclerosis. AB - Ischaemic stroke ranks among the most important causes of death and disability in developed countries. Abnormal lipid metabolism is among the several factors that have a role in the pathogenesis of atherosclerosis. We hypothesize that the decreased resistance of low-density lipoproteins against oxidative stress is an independent risk factor for cerebral atherosclerosis and suggest testing this hypothesis by ultrasonographic evaluation of the carotid artery and correlating this finding to plasma values of compounds that play a role in lipid metabolism. By measuring the oxidative resistance of low-density of lipoprotein the risk for ischaemic stroke can be predicted. PMID- 11399121 TI - The problem of Gulf War syndrome. AB - Following a war with widespread attention to and concern over the potential for numerous biological and chemical warfare exposures, some Gulf War veterans returned home and developed various illnesses. Although some of these illnesses are readily diagnosable, the so-called Gulf War syndrome has remained a controversial and nebulous diagnosis. It is characterized by multiple, subjective symptoms, and by a lack of objective pathology. To date, the search for a single disease entity and a biological model to explain this illness has been unsuccessful. Wars have long affected the health of veterans in multiple ways, and a single disease entity is not likely as a viable explanation for these outcomes. Given the nature of the illness, and its overlap with many other controversial chronic illnesses, we suggest that the biopsychosocial model may provide a better solution to this diagnostic conundrum. PMID- 11399122 TI - Aromatase, adiposity, aging and disease. The hypogonadal-metabolic-atherogenic disease and aging connection. AB - In males, aging, health and disease are processes that occur over physiologic time and involve a cascade of hormonal, biochemical and physiological changes that accompany the down-regulation of the hypothalamic-anterior pituitary testicular axis. As aging progresses there are relative increases of body fat and decreases in muscle mass. The increased adipose tissue mass is associated with the production of a number of newly generated factors. These include aromatase, leptin, PAI-1, insulin resistance, and the dyslipidemias, all of which can lead to tissue damage. Fatty tissue becomes the focal point for study as it represents the intersection between energy storage and mobilization. The increase in adipose tissue is associated with an increase in the enzyme aromatase that converts testosterone to estradiol and leads to diminished testosterone levels that favor the preferential deposition of visceral fat. As the total body fat mass increases, hormone resistance develops for leptin and insulin. Increasing leptin fails to prevent weight gain and the hypogonadal-obesity cycle ensues causing further visceral obesity and insulin resistance. The progressive insulin resistance leads to a high triglyceride-low HDL pattern of dyslipidemia and increased cardiovascular risk. All of these factors eventually contribute to the CHAOS Complex: coronary disease, hypertension, adult-onset diabetes mellitus, obesity and/or stroke as permanent changes unfold. Other consequences of the chronic hypogonadal state include osteopenia, extreme fatigue, depression, insomnia, loss of aggressiveness and erectile dysfunction all of which develop over variable periods of time. PMID- 11399123 TI - Kinetics and mechanisms of hepatic acute phase response to subtotal partial hepatectomy and cultural impact on environmental hepatic end-stage liver injury in the homeless. AB - Intoxication and liver damage induced by carbon tetrachloride (CCl(4)), aflatoxin B1, diabetes, and subtotal partial hepatectomy (PH(90)) in rats in which approximately 90% of the total hepatic tissue mass is surgically removed produces an acute-phase response (APR) whose initial stage prior to regression closely mimics the APRs associated with the life-threatening hepatic failure seen in the homeless. Rats treated by PH(90)were either healthy, CCl(4)-intoxicated, diabetic, or alflatoxin B1 (AFB1) intoxicated to the point of 75% liver insufficiency. It is well documented that high rates of mortality following PH(90)in aseptic rats could be minimized by supplementing drinking water with 20% glucose, organic components of L-15 medium and housing animals in cages maintained at 33-35;C. Aseptic rats showed a mild 20-30% decrease in APR proteins during the first 4-5 days following PH(90), while a maximal APR was noted 9-12 days post PH(90)and lasted for ~30 days when it returned to values close to those of healthy controls. This delay in hepatic APR of the remnant caudate lobe favoured replacement of lost basophilic clumps and ribosomes. The newly synthesized ribosomes of the nascent hepatocytes quantitatively maintained the APR signals of the injured caudate hepatocytes, and biosynthesized and released a typical spectrum of APR proteins. We suggest that massively injured liver has decoded an already stored and irreversible DNA-biochemical sequence of events in which priority is given to recovery of lost tissues by delaying an APR response to injury. In PH(90)of diabetic and CCl(4)-intoxicated rats, the hepatic dual functions of regeneration and APR processes associated with intoxication initiated catabolic signals, created a heavy metabolic burden on the remnant caudate lobe leading to higher rates of mortality. APR of healthy rats to AFB1 parallels that of alpha-amanitin-induced intoxication. Similarly, within shorter time scale proportional to the severity of surgery, livers undergoing 75% partially hepatectomy (PH(75)) delayed both the onset and regression of APR. We are therefore led to believe that approaches other than liver transplantation should be considered as viable alternatives in the treatment of various acute and chronic liver diseases to avoid rejection and retransplantation. Scarcity of cadaveric liver has forced the medical community to investigate xenotransplantation with its unknown risks. Concomitantly, it is suggested that in view of the incalculable risks of indifference, the homeless must receive much improved medical care as we have found that two-dimensional immunoelectrophoretic assay of their serum is indicative of acute and chronic liver injury. The scientific and moral interrelationships of related matters are illuminated. PMID- 11399124 TI - Cancer genesis: stem tumour cells as an MHC-null/HSP70 - very high 'primordial self' escaping both MHC-restricted and MHC-non- restricted immunesurveillance. AB - I have previously assumed that in tumours there are stem cells, that owing to the morphophysiologic properties shared with embryonal cells I have defined as 'para embryonal' cells (PECs). Owing to a blocking mutation, PECs might be able to express only the genic program upstream from the block, but not that downstream. As a consequence, PECs might lack in genic differentiated products, such as MHC molecules, and might be very rich in primitive genic products, such as HSP70 molecules. Like embryonal cells, PECs might carry out induction on adjoining hyperplastic cells, thus transforming them, only phenotypically, into 'differentiated para-embryonal cells' (DPECs), endowed with both MHC and HSP70 molecules. In such a way, nuclei of MHC-non-expressing/HSP70-high expressing stem tumour cells might be surrounded by layers of MHC-expressing/HSP70-expressing non stem tumour cells. Such a structural tumour organization, actually found by C. Cordon Cardo et al. with regard to the MHC molecule expression, might be responsible for interference phenomena versus the MHC-non-restricted immune cells, such as macrophages and NK cells. So, these cells, the only potentially able to recognize and eliminate MHC-non-expressing stem tumour cells (PECs), might spare them, thus rendering cancer a biological process without any natural immunological solution. Now, I would like to theorically demonstrate that cancer might be a process without immunological solution from the very beginning: the first stem tumour cell might be tolerated as a sort of 'primordial self' because of its MHC-null/HSP70-very high phenotype, recognizable by neither the MHC restricted nor the MHC-non-restricted immunesurveillance systems of the host. Possible biological roles of HSP70 molecules might account for the immunesurveillance escape of stem tumour cells. Existence of these cells appears to be confirmed by the recent experiments of immunotherapy with autologous tumour specific HSPs carried out by P. K. Srivastava; moreover, their 'self' nature appears to be confirmed by the most recent experiments of compatible bone marrow allograft carried out by A. M. Carella. On this ground, the main steps for a resolutive antitumour immunotherapy are proposed. PMID- 11399125 TI - Iron, infection and sudden infant death. AB - Three risk factors for sudden infant death syndrome are well established: maternal smoking, prone sleeping position and non-breast feeding. Two additional risk factors have been proposed: microbial infection in the gastrointestinal or respiratory tracts and iron loading. This review endeavors to integrate these five disparate factors into a unifying concept. PMID- 11399126 TI - Dose-years as an improved index of cumulative tobacco smoke exposure. AB - In assessing the link between tobacco smoking and disease, it is important to determine longterm, cumulative exposure to tobacco smoke as accurately as possible. Conventional methods of assessing exposure to tobacco smoke each have intrinsic limitations. Self-reporting of tobacco use, and the conversion of this data to pack-years, can be prone to error due to individuals wishing to conceal smoking habits, inaccurate reporting of daily cigarette consumptions or years of smoking, and failure to take into account the variation that exists in inter individual smoking experiences. Measurement of cotinine, a major metabolite of nicotine in humans, is a reliable method of monitoring recent doses of tobacco smoke exposure. Cotinine concentrations, however, may remain stable in smokers over the longer term. Therefore, dose-years, and more specifically cotinine years, may represent an improved index of cumulative tobacco smoke exposure. PMID- 11399127 TI - Sperm capacitation and primary sex ratio. AB - The observations of many years enabled determination of maternal factors which influence the sex of progeny. Factors on side of the father have long been neglected, even though the sex of progeny depends on the sex chromosome in fertilizing spermatozoon. The data obtained by penetration method 'hamster oocyte - human sperm' show differences in the proportion of sperm cells, depending on the method of capacitation used. On this basis the hypothesis has been formulated that the factor which precludes the proportion of sex at fertilization may be different time of capacitation of sperm cells. The cause of the phenomenon remains unclear. PMID- 11399128 TI - Can 'human rights' and 'women' go together? PMID- 11399129 TI - Can improvements in breast-feeding practices reduce neonatal mortality in developing countries? AB - OBJECTIVE: to review the literature on the relationship between breast-feeding practices in the first month of life and neonatal mortality. METHODS: Medline and Cochrane databases were searched using the keywords breastfeeding, and neonatal mortality, supplemented with additional searches using the keywords developing countries, colostrum, infant feeding and infant mortality, hypoglaecemia, hypothermia, breastfeeding practices, and suckling. FINDINGS: breast feeding helps prevent hypothermia and hypoglycaemia in newborn babies, which are contributory causes of early neonatal deaths especially among low birth weight and premature babies. During the late neonatal period, most deaths in developing countries are due to infections such as sepsis, acute respiratory tract infection, meningitis, omphalitis and diarrhoea. Feeding colostrum and breast feeding, especially exclusive breast feeding, protects against such deaths. KEY CONCLUSIONS AND IMPLICATIONS FOR PRACTICE: in most developing countries, nearly all women breast feed in the first month of life, but often breast feeding is delayed beyond the first hour after birth, and exclusive breast feeding is not usually practised. Policies and training of staff of maternity centres and hospitals can encourage early initiation of breast feeding and exclusive breast feeding. Midwives can support community-based efforts to support exclusive breast feeding. Breast feeding plays an important role in reducing neonatal mortality and should be strongly emphasised by programmes attempting to reduce neonatal mortality. PMID- 11399130 TI - A review of the literature on the midwife and community-based maternity care. AB - OBJECTIVE: to review the UK literature relating to community-based maternity care. DESIGN: all UK research studies published between 1970 and 1998 relating to community-based maternity care were included. Searches were made via a number of electronic databases using defined search terms. All papers included were independently reviewed by a minimum of two researchers. Study findings were tabulated using a pro-forma. Findings are summarised in this paper. FINDINGS: a total of 241 papers were deemed to meet all inclusion criteria. The majority of studies used descriptive methods with only 11 papers reporting findings from randomised controlled trials. Findings are reported relating to clinical outcomes, the care process and the views of women and health professionals. CONCLUSION AND POLICY IMPLICATIONS: the overall quality of the evidence in the papers reviewed was very mixed. What limited evidence there is suggests that, for the majority of women, care in community settings is as safe and as acceptable to women as care provided in hospital. Despite a large volume of literature, the amount that is known about midwives' contribution to care, and what women think about it, is limited. There is a need for controlled studies to compare outcomes for different patterns of care and for well-designed observational studies to provide information on the care process. PMID- 11399131 TI - The lived experiences of autonomous Angolan midwives working in midwifery-led, maternity units. AB - OBJECTIVE: to describe the lived experiences of autonomous midwives working in Angolan midwifery-led maternity units. DESIGN: a qualitative approach using semi structured, audiotaped interviews, in Portugese. Data were analysed in a six-step process. SETTING: three midwifery-led maternity units in the most densely populated suburbs in the capital of Angola, Luanda. The average number of deliveries per unit was 2500 per year. PARTICIPANTS: 11 midwives from the three maternity units. FINDINGS: four main areas emerged: society/culture, significant others, personal self and professional self. Sub-areas, concepts and supporting statements were defined in each area. KEY CONCLUSIONS: the midwives served within a population living in rough circumstances but which maintained strong traditional roots. The midwives did not support homebirths, but did assist when needed. The midwives described their professional role as a 'calling', which was very independent. Cure, was considered more important than care, and strong emotions were expressed when discussing cases of failure. The partograph was viewed as an important instrument and continuous learning as crucial in their role as autonomous midwives. IMPLICATIONS FOR PRACTICE: the model of a midwifery led delivery unit described in this study may be used in other countries facing the same problems as Angola. Difficulties concerning transfer should be seriously considered as well as adequate education for the midwives. A pre-requisite in order for peripheral maternity units to have any impact on maternal morbidity and mortality, is a well-organised first-referral level. PMID- 11399132 TI - A qualitative study of the perceptions of Hong Kong Chinese women during caesarean section under regional anaesthesia. AB - AIM: to explore Hong Kong Chinese women's experiences and perceptions of elective caesarean section performed under regional anaesthesia. DESIGN: an exploratory study using a qualitative approach. Data were collected by tape-recorded in-depth interviews two to five days after caesarean section. PARTICIPANTS: a purposive sample of 18 Hong Kong Chinese women having an uncomplicated, elective caesarean section under successful regional anaesthesia. SETTING: a post-caesarean ward of a University affiliated District General Hospital in Hong Kong. KEY FINDINGS: the three categories relating to the overall theme of Consciousness during Surgery were: Interacting with others, Experience of birthing, and Awareness of the environment. IMPLICATIONS FOR PRACTICE: the experiences of women, related to the general theme and categories cited above, affected their feelings of security and fulfillment. On the whole, women were satisfied with the regional anaesthesia, preferring to remain conscious throughout the surgery. They appreciated being able to interact with others and listening to music, but needed information regarding what was happening. Many were distressed by the sight and noise of surgical instruments, the narrowness of the operating table, the operating theatre lamps and the coldness of the theatre. Recommendations for practice are made relating to these issues. PMID- 11399133 TI - Antenatal, delivery and postnatal comparisons of maternal satisfaction with two pilot Changing Childbirth schemes compared with a traditional model of care. AB - OBJECTIVE: to investigate maternal satisfaction with two pilot schemes based on the Changing Childbirth initiative (DoH 1993) and to compare this with a traditional model of care. In addition, a limited number of clinical outcome measures were also assessed. DESIGN: a retrospective between-group design was used. Questionnaire data were collected from three groups (two pilot and one control) about the antenatal, labour and postnatal periods to establish both satisfaction with key objectives of the Changing Childbirth initiative (DoH 1993), and basic clinical outcomes. SETTING: a large Trust (see definition in main article) in Central England, that covered a wide range of socio-economic strata. PARTICIPANTS: the two pilot groups comprised 112 and 103 women respectively and were randomly drawn from GP practices within the Trust's catchment area. The third group of 118 women (Control) was selected from the Trust's obstetric unit. Women at high obstetric risk were excluded from this study. MEASUREMENT: a five-part questionnaire was devised that covered: (1) preferences for type of care, health-care professional, venue etc; (2) details of antenatal care provision and the participants' satisfaction with this; (3) labour, including clinical outcomes, labour and birth details and satisfaction with care; (4) postnatal care information, including satisfaction scores; and (5) information and advice given throughout the ante, delivery and postnatal periods and satisfaction with this. The questionnaires were administered six weeks postnatally. FINDINGS: although the two pilot groups had been set up to follow a one-to-one midwifery care model, the second group naturally evolved into providing care from within a small group of midwives. This variation did not lead to any differences in any of the outcome measures. The women in the obstetrician led group were not dissatisfied with the care, information and treatment they received, but they were significantly less satisfied than either of the two pilot groups. The pilot groups also rated more highly the information and choice that they had, and felt that the midwives acted as partners in the process. These findings applied to the antenatal, delivery and postnatal periods. No differences in clinical outcomes were observed between the groups. CONCLUSIONS: midwifery-led care was much preferred to obstetrician-led care and did not lead to any deficits in clinical outcomes. The pilot scheme that adapted the initiative into small group provision showed no reduction in satisfaction levels or other outcome measures. Since burn-out and stress have been identified as features of one-to one midwifery provision, this model might have potential for offsetting this problem, while still maintaining the spirit of the Changing Childbirth policy. From the perspective of maternal well-being, both physical and psychological, the initiative reported here appears to have been highly successful. PMID- 11399134 TI - An exploration of midwives' and obstetricians' knowledge of genetic screening in pregnancy and their perception of appropriate counselling. AB - OBJECTIVE: to describe levels of knowledge relevant to genetic screening in a sample of midwives and obstetricians in the late 1990s, and to compare these with those found by Smith et al. in London and Wales, and reported in 1994; to describe health professionals' perceptions of an appropriate counselling process relating to genetic screening in comparison with recognised good practice; and to consider links between knowledge and perceptions of the counselling process. DESIGN: a questionnaire study, including responses to a counselling scenario. PARTICIPANTS AND SETTING: responses were obtained from 81 midwives and obstetricians from maternity services in the North West of England. FINDINGS: knowledge about procedures was very good, but knowledge about the probability of genetic abnormality was relatively poor, and respondents overestimated the efficacy and usefulness of tests. These findings were similar to those of Smith et al. (1994), suggesting that they are a good reflection of the national picture. In terms of reported information-giving and counselling practice, there were some respondents who would not check the woman's understanding of her baby's risk of a genetic abnormality, or the risk of a false result. Some respondents would give information based on their own (necessarily limited) experience, rather than national statistics, and give advice based on the choices they themselves would make. These findings reflect earlier, basic research on people's understanding of probabilistic information. IMPLICATIONS: the findings of this study, together with those of earlier work cited, suggest a need for education and training which includes a specific focus on biases in understanding this type of probabilistic information. They also raise the question as to why tests which provide probabilistic data have been introduced without consideration of the known problems in understanding such information. PMID- 11399135 TI - A literature review on the antiphospholipid syndrome and the effect on childbearing. AB - OBJECTIVE: to review the literature on antiphospholipid antibodies and their significance to midwifery practice. METHOD: databases using the keywords anticardiolipin antibodies, antiphospholipid antibodies, lupus antibodies, antiphospholipid syndrome, systemic lupus erythematosis, pregnancy loss, pre eclampsia were searched, 548 articles were generated and 52 were used in the review. FINDINGS: anticardiolipin antibodies are detrimental to successful pregnancy being implicated in failure of implantation, loss of the embryo and the fetus. There is also a risk of prematurity, intrauterine growth retardation and neonatal embolism. The risks to the mother are of pre-eclampsia and embolic complications. If diagnosed and treated early in pregnancy with low-dose aspirin and subcutaneous heparin the outlook for a successful pregnancy is much improved. KEY CONCLUSIONS: much research remains to be undertaken. Researchers need to standardise their criteria so that findings are comparable and larger sample groups are necessary. IMPLICATIONS FOR PRACTICE: antiphospholipid syndrome is an important disorder and a preventable cause of pregnancy loss. Midwives need to encourage women to persevere with therapy to preserve pregnancy and maybe prevent pre-eclampsia or thrombosis. When pregnancies are lost one of the hardest things is when the parents cannot understand why. Midwives need to understand the syndrome to collaborate with doctors in explaining it and supporting grieving parents. PMID- 11399136 TI - A pilot study to assess the viability of a randomised controlled trial of methods of supplementary feeding of breast-fed pre-term babies. AB - OBJECTIVES: to compare the impact of two methods of supplementary feeding of pre term babies (bottle vs cup) on subsequent breast feeding and to assess the feasibility of using a randomised controlled trial (RCT) to investigate the topic. DESIGN AND METHOD: small scale prospective RCT. Data on breast feeding, as defined as the exclusive method of feeding, were collected. A range of relevant bio-data was also collected and their impact on breast feeding assessed. SETTING: a special care baby unit in a District General Hospital in the UK. PARTICIPANTS: over a three-month period, all pre-term babies (32-37 weeks' gestation) who fulfilled the inclusion criteria and has been born to mothers who had expressed a pre-partum desire to breast feed, who had consented to take part, were included (n=14). PROCEDURE: the eligible babies were randomly allocated to supplementary feeding of breast milk, via either a cup or a bottle. Whether or not the baby was being breast fed at discharge was noted. FINDINGS: the study suggested that this RCT framework is a viable method of investigating baby feeding. Because of the small-scale nature of the project, the actual database must be treated with extreme caution. No significant differences were found between the two groups in terms of breast feeding. However, the mothers reported high levels of support and also the breast-feeding rates were above the national averages. These two findings could have contributed to the non-significant results observed in this analysis. CONCLUSIONS: if the present findings could be supported by further research, then the non-significant results relating method of supplementary feeds to subsequent breast feeding could be explained by reference to three factors. Firstly, there is, in fact, no real effect of method of supplementary feeding and subsequent breast feeding; secondly, the method adopted differed from existing research and thus may be expected to produce non-corroborative results; and finally, the overall levels of breast feeding within the Unit generally were higher than the national average. The relevance of the RCT for investigating this subject is also discussed with reference to the present data set. Further experimental work to develop these ideas and to identify causal links is required. PMID- 11399138 TI - The role of different protein components from the Haemophilus ducreyi cytolethal distending toxin in the generation of cell toxicity. AB - Cytolethal distending toxin of Haemophilus ducreyi (HdCDT) is a multicomponent toxin, encoded by an operon consisting of three genes, cdtABC. To investigate the role of the individual products in generation of toxicity, recombinant plasmids were constructed allowing expression of each of the genes individually or in different combinations in Escherichia coli and Vibrio cholerae. Expression of all three genes (cdtABC) was necessary to generate toxicity on cells, and no activity was obtained using combinations in which only one or two of the genes were expressed. Of the individual gene products, the CdtA was shown to exist in two forms with an MW of 23 and 17 kDa, respectively. The CdtB protein alone resulted in DNase activity. CdtC purified from both toxic and non-toxic extracts (from strains expressing cdtCAB and cdtC, respectively) had a molecular weight of about 20 kDa and reacted with a CdtC-specific monoclonal antibody. However, the protein isoelectric point (pI) of CdtC from toxic preparations was about 1.5 pH units more basic than from non-toxic ones. Both forms were immunogenic giving rise to toxin-neutralizing antibodies. Toxicity was reconstructed by combining non-toxic cell sonicates from E. coli, expressing CdtA, CdtB and CdtC proteins individually. Only combinations including all three products gave toxicity, indicating that all are actively involved in the generation of toxic activity on cells. The reconstruction resulted in a 1.5 pH unit shift in the PI of CdtC, making it identical to that of the protein isolated from bacteria expressing cdtABC. The results showed that the CdtB component produces DNase activity, but cell toxicity depends on the involvement of the other two components of CDT and is associated with absorption of all three proteins by HEp-2 cells. PMID- 11399139 TI - Mast cell density and substance P-like immunoreactivity during the initiation and progression of lung lesions in ovine Mannheimia (Pasteurella) haemolytica pneumonia. AB - To determine the density of mast cells (MCs) and the extent of substance P (SP) immunoreactivity during initiation and progression of pneumonic pasteurellosis (PP), 18 lambs were inoculated intrabronchially with Mannheimia (Pasteurella) haemolytica or saline, and lung tissue was collected at 1, 15 and 45 days post inoculation (n=3, each group). Additionally, the left (non-inoculated) contralateral lungs in bacteria-inoculated animals were collected as controls. At 1 day after bacterial inoculation the lungs had typical M. haemolytica lesions. These pneumonic lesions had fewer numbers of MCs and reduced histamine content. Macrophages infiltrating some of the inflamed areas were strongly immunoreactive for SP. At 15 days, MCs remained scarce at sites where lung damage persisted, i.e. pyogranulomatous foci, but were increased in number in areas of interstitial damage. Pulmonary ganglion neurons were strongly immunoreactive for SP. By 45 days the fibrosing changes became more defined as pleural fibrosis, fibrosing alveolitis, alveolar epithelial hyperplasia and bronchiolitis obliterans. These lungs had increased numbers of MCs, but histamine content was not different from saline- and non-inoculated left lungs. Substance P immunoreactivity occurred only in nerves and was scarce and mild. This work demonstrates that MC density decreases initially with PP, but increases with progression of PP. SP fibres tend to be decreased during the initiation and at 45 days of PP, but other cells, such as macrophages and neuronal ganglion cells, produce substance P during progression of PP and thereby constitute an additional source of substance P. PMID- 11399140 TI - Differential expression of ompA, ompB, pyk, nlpD and Cpn0585 genes between normal and interferon-gamma treated cultures of Chlamydia pneumoniae. AB - A common feature of many chlamydial infections is that they are often asymptomatic and may persist for long periods of time if left untreated. In addition to the well recognized lytic stage of the chlamydial developmental cycle, evidence is now emerging to support a persistent phase in the cycle in which the reticulate bodies are morphologically abnormal, viable but non infectious and presumably also have altered gene expression patterns. We used an RT-PCR approach to study the differential levels of gene transcription for 14 genes (16SrRNA, ompA, ompB, omcB, 76 kDa, gseA, pmp1, gltX, hsp60, yaeT, pyk, nlpD, Cpn0585, Cpn1046) between normal and IFN-gamma treated Chlamydia pneumoniae cell cultures. Even though the level of morphologically abnormal reticulate bodies in our IFN-gamma treated cultures was low (approximately 10% morphologically discernible, although presumably a larger percentage were in the persistent state but not yet morphologically altered) we identified five genes (ompA, ompB, pyk, nlpD, Cpn0585) that were clearly upregulated when compared to normal cultures. This gene transcript profile may be characteristic of a general stress state in Chlamydia, induced by IFN-gamma treatment in this case, but perhaps more widely induced in other in vitro and in vivo situations. PMID- 11399141 TI - Lipopolysaccharide enhances cytolysis and inflammatory cytokine induction in bovine alveolar macrophages exposed to Pasteurella (Mannheimia) haemolytica leukotoxin. AB - Pasteurella (Mannheimia) haemolytica leukotoxin (Lkt) and lipopolysaccharide (LPS) are the primary virulence factors contributing to the pathogenesis of lung injury in bovine pneumonic pasteurellosis. Previous studies have characterized in vitro responses of bovine alveolar macrophages (AMs) to Lkt and LPS. Activation of AMs with Lkt or LPS causes induction of proinflammatory cytokines, and Lkt causes cytolysis of AMs at higher concentrations. Since AMs are exposed to both of these bacterial virulence factors during disease, previous studies may have underestimated the possibility of functional interactions between Lkt and LPS. The purpose of this study was to characterize the effect of simultaneous exposure to both Lkt and LPS on AM cytolysis and proinflammatory cytokine expression. Using cellular leakage of lactate dehydrogenase as an indirect measure of cytolysis, we studied AM responses to Lkt alone, LPS alone and Lkt+LPS. We found that 80-200 pg/ml LPS, which does not itself cause cytolysis, synergistically enhanced the cytolysis induced by 2-5 Lkt units (LU)/ml Lkt. Northern blot analysis demonstrated that synergism between Lkt and LPS resulted in increased levels of IL-8 mRNA, and that the kinetic patterns of TNF-alpha and IL-8 mRNA expression induced by Lkt+LPS differed from those induced by each agent separately. Finally, the WEHI 164 (clone 13) bioassay was used to show that Lkt/LPS synergism resulted in enhanced secretion of biologically active TNF alpha. These results provide direct evidence of synergism between Lkt and LPS in AM cytolysis and inflammatory cytokine expression. Additional studies to characterize the molecular basis of this phenomenon are indicated. PMID- 11399142 TI - Analysis of the effect of changing environmental conditions on the expression patterns of exported surface-associated proteins of the oral pathogen Actinobacillus actinomycetemcomitans. AB - Actinobacillus actinomycetemcomitans has been specifically implicated in the aetiology of one or more of the periodontal diseases, conditions in which inflammation of the gums is associated with destruction of the alveolar bone supporting the teeth. In these diseases there is loss of attachment of the gums (gingivae) to the teeth forming a periodontal pocket. The microenvironment of this pocket is extremely complex and it is likely that there will be substantial variation in the environmental conditions operating in this habitat. The aim of the current investigation was to study the effect of disease-relevant environmental factors on the production and release of secreted surface- associated proteins of A. actinomycetemcomitans. These secreted proteins contain many of the virulence determinants of this organism. A range of environmental conditions were investigated: growth in a CO(2)-enriched aerobic atmosphere vs anaerobic growth, presence of serum or blood, biofilm vs planktonic mode of growth and iron depletion. Differential expression of a number of the secreted surface-associated proteins was observed under different growth conditions and these included the glycolytic enzyme triose phosphate isomerase. An ability to adapt to prevailing environmental conditions may facilitate the survival of the organism in the changing microIenvironment of the periodontal pocket. PMID- 11399144 TI - Molecular analysis of phylogenetic relationships among Myrmecophytic macaranga species (Euphorbiaceae). AB - Many species of the paleotropical pioneer tree genus Macaranga Thou. (Euphorbiaceae) live in association with ants. Various types of mutualistic interactions exist, ranging from the attraction of unspecific ant visitors to obligate myrmecophytism. In the latter, nesting space and food bodies are exchanged for protection by highly specific ant partners (mainly species of the myrmicine genus Crematogaster). As a first step toward elucidating the coevolution of ant-plant interactions in the Macaranga-Crematogaster system, we have initiated a molecular investigation of the plant partners' phylogeny. Nuclear ribosomal DNA internal transcribed spacer (ITS) sequences were analyzed for 73 accessions from 47 Macaranga species, representing 17 sections or informally described species groups. Three accessions from the putative sister taxon Mallotus Lour, were included as outgroups. Cladograms of the ITS data revealed Macaranga to be nested within Mallotus. ITS sequences are highly similar within section Pachystemon s.str., suggesting a relatively recent and rapid radiation of obligate myrmecophytes within this section. Forty-three accessions, mainly of ant-inhabited species, were additionally investigated by random amplified polymorphic DNA (RAPD) and microsatellite-primed PCR (MP-PCR) techniques. Phenetic analysis of RAPD and MP-PCR banding profiles generally confirmed the ITS results. Best resolutions for individual clades were obtained when ITS and RAPD/MP-PCR data were combined into a single matrix and analyzed phenetically. The combined analysis suggests multiple (four) rather than a single evolutionary origin of myrmecophytism, at least one reversal from obligate myrmecophytism to nonmyrmecophytism, and one loss of mutualistic specifity. PMID- 11399145 TI - One-step PCR amplification of complete arthropod mitochondrial genomes. AB - A new PCR primer set which enables one-step amplification of complete arthropod mitochondrial genomes was designed from two conserved 16S rDNA regions for the long PCR technique. For this purpose, partial 16S rDNAs amplified with universal primers 16SA and 16SB were newly sequenced from six representative arthropods: Armadillidium vulgare and Macrobrachium nipponense (Crustacea), Anopheles sinensis (Insecta), Lithobius forficatus and Megaphyllum sp. (Myriapoda), and Limulus polyphemus (Chelicerata). The genomic locations of two new primers, HPK16Saa and HPK16Sbb, correspond to positions 13314-13345 and 12951-12984, respectively, in the Drosophila yakuba mitochondrial genome. The usefulness of the primer set was experimentally examined and confirmed with five of the representative arthropods, except for A. vulgare, which has a linearized mitochondrial genome. With this set, therefore, we could easily and rapidly amplify complete mitochondrial genomes with small amounts of arthropod DNA. Although the primers suggested here were examined only with arthropod groups, a possibility of successful application to other invertebrates is very high, since the high degree of sequence conservation is shown on the primer sites in other invertebrates. Thus, this primer set can serve various research fields, such as molecular evolution, population genetics, and molecular phylogenetics based on DNA sequences, RFLP, and gene rearrangement of mitochondrial genomes in arthropods and other invertebrates. PMID- 11399146 TI - Phylogeny of the Celastraceae inferred from 26S nuclear ribosomal DNA, phytochrome B, rbcL, atpB, and morphology. AB - Phylogenetic relationships within Celastraceae (spindle-tree family) were inferred from nucleotide sequence characters from the 5' end of 26S nuclear ribosomal DNA (including expansion segments D1-D3; 84 species sampled), phytochrome B (58 species), rbcL (31 species), atpB (23 species), and morphology (94 species). Among taxa of questionable affinity, Forsellesia is a member of Crossosomataceae, and Goupia is excluded from Celastraceae. However, Brexia, Canotia, Lepuropetalon, Parnassia, Siphonodon, and Stackhousiaceae are supported as members of Celastraceae. Gymnosporia and Tricerma are distinct from Maytenus, Cassine is supported as distinct from Elaeodendron, and Dicarpellum is distinct from Salacia. Catha, Maytenus, and Pristimera are not resolved as natural genera. Hippocrateaceae (including Plagiopteron and Lophopetalum) are a clade nested within a paraphyletic Celastraceae. These data also suggest that the Loesener's classification of Celastraceae sensu stricto and Halle's classification of Hippocrateaceae are artificial. The diversification of the fruit and aril within Celastraceae appears to be complex, with multiple origins of most fruit and aril forms. PMID- 11399147 TI - Evolutionary relationships of trichostrongyloid nematodes (Strongylida) inferred from ribosomal DNA sequence data. AB - The evolutionary relationships of 21 species of trichostrongyloid nematodes were determined by use of sequence data of the second internal transcribed spacer of the ribosomal DNA aligned according to secondary structure information. Irrespective of the method of analysis used, the topologies of the phylogenetic trees derived from the molecular data differed with respect to all four hypotheses proposed previously for the evolutionary relationships of the different subfamilies within the Trichostrongylidae based on morphological data. Thus, the molecular data set did not resolve the conflict between the four previous proposals for the subfamilial relationships. Nonetheless, all trees derived from the molecular data showed strong support for the exclusion of the genera Filarinema and Amidostomum from the clade containing the species within the family Trichostrongylidae. This represents a major difference from the most recent proposal of the systematics of the Trichostrongyloidea in which these two genera were included within the Trichostrongylidae. Therefore, the molecular data support an earlier systematic framework in which Filarinema and Amidostomum were considered to be sister groups of the Trichostrongyloidea. PMID- 11399148 TI - Phylogeny and biogeography of the flowering plant genus Styrax (Styracaceae) based on chloroplast DNA restriction sites and DNA sequences of the internal transcribed spacer region. AB - Phylogenetic relationships within the flowering plant genus Styrax were investigated with DNA sequence data from the internal transcribed spacer (ITS) region of nuclear ribosomal DNA (nrDNA) and with chloroplast DNA restriction site data from the genes trnK, rpoC1, and rpoC2. The data sets from each genome were analyzed separately and in combination with parsimony methods. The results strongly support the monophyly of each of the four series of the genus but provide little phylogenetic resolution among them. Reticulate evolution may at least partly explain discordance between the molecular phylogenetic estimates and a prior morphological estimate within series Cyrta. The historical biogeography of the genus was inferred with unweighted parsimony character optimization of trees recovered from a combined ITS and morphological data set, after a series of combinability tests for data set congruence was conducted. The results are consistent with the fossil record in supporting a Eurasian origin of Styrax. The nested phylogenetic position of the South American members of the genus within those from southern North America and Eurasia suggests that the boreotropics hypothesis best explains the amphi-Pacific tropical disjunct distribution occurring within section Valvatae. The pattern of relationship recovered among the species of section Styrax ((western North America + western Eurasia) (eastern North America + eastern Eurasia)) is rare among north-temperate Tertiary forest relicts. The monophyly of the group of species from western North America and western Eurasia provides qualified support for the Madrean-Tethyan hypothesis, which posits a Tertiary floristic connection among the semiarid regions in which these taxa occur. A single vicariance event between eastern Asia and eastern North America accounts for the pattern of relationship among intercontinental disjuncts in series Cyrta. PMID- 11399149 TI - Phylogenetic utility of the nuclear gene malate synthase in the palm family (Arecaceae). AB - There is currently a shortage of DNA regions known to be useful for phylogenetic research in palms (Arecaceae). We report the development and use of primers for amplifying and sequencing regions of the nuclear gene malate synthase. In palms the gene appears to be single-copy, with exon regions that are phylogenetically informative within the family. We constructed a phylogeny of 45 palms and five outgroup taxa using 428 bp of malate synthase exon regions. We found that some major clades within the family were recovered, but there was a lack of resolution among the genera in subfamilies Arecoideae, Ceroxyloideae, Coryphoideae, and Phytelephantoideae. In a second analysis, malate synthase exon regions totaling 1002 bp were sequenced for 16 palms and two outgroup taxa. There was increased bootstrap support for some groups and for the placement of the monotypic genus Nypa as sister to the rest of the family. A comparison with data sets from noncoding regions of the chloroplast genome indicates that malate synthase sequences are more variable and potentially contain more phylogenetic information. We found no evidence of multiple copies of the malate synthase gene in palm genomes. PMID- 11399150 TI - Phylogenetic utility of the glycerol-3-phosphate acyltransferase gene: evolution and implications in Paeonia (Paeoniaceae). AB - The nuclear-encoded chloroplast-expressed glycerol-3-phosphate acyltransferase (GPAT) gene has been found to be single-copy in a number of angiosperm families. In this study we investigated the phylogenetic utility of the GPAT gene at the interspecific level using the genus Paeonia (Paeoniaceae) as an example. An approximately 2.3- to 2.6-kb fragment of the GPAT gene, containing a large intron of more than 2 kb, was amplified, cloned, and sequenced from 19 accessions representing 13 Paeonia species. The GPAT gene phylogeny inferred by parsimony analysis supported interspecific relationships that were previously unresolved, suggesting that large introns of low-copy nuclear genes are particularly informative in the resolution of close relationships at low taxonomic levels. Whereas the GPAT phylogeny is largely congruent with the previous phylogenetic hypothesis of Paeonia, it shows a significant discordance involving the paraphyly of section Paeonia. Given evidence of an ancient duplication and the subsequent silencing of one GPAT locus in P. anomala, this discordance is most likely the result of paralogy. Two distinct genomic clones containing partial GPAT genes were isolated from P. anomala. The GPAT sequence from one clone corresponded to the functional copy of the gene, and the second genomic clone was determined to contain a GPAT pseudogene. The insertion of a retrotransposon in an intron of this pseudogene may have been responsible for the silencing of this GPAT locus in P. anomala. This study suggests that, although it is unlikely that universal nuclear gene markers free from paralogy are usually available, low-copy nuclear genes can be very useful in plant phylogenetic reconstruction, especially at low taxonomic levels, as long as the evolutionary dynamics of the genes are carefully examined. PMID- 11399151 TI - Phylogeny of the Trichostrongylina (Nematoda) inferred from 28S rDNA sequences. AB - We produced a molecular phylogeny of species within the order Strongylida (bursate nematodes) using the D1 and D2 domains of 28S rDNA, with 23 new sequences for each domain. A first analysis using Caenorhabditis elegans as an outgroup produced a tree with low resolution in which three taxa (Dictyocaulus filaria, Dictyocaulus noerneri, and Metastrongylus pudendotectus) showed highly divergent sequences. In a second analysis, these three species and C. elegans were removed and an Ancylostomatina, Bunostomum trigonocephalum, was chosen (on the basis of previous morphological analyses) as the outgroup for an analysis of the phylogenetic relationships between and within the Strongylina (strongyles) and Trichostrongylina (trichostrongyles). A very robust tree was obtained. The Trichostrongylina were monophyletic, but the Strongylina were paraphyletic, though this requires confirmation. Within the Trichostrongylina, the three superfamilies defined from morphological characters are confirmed, with the Trichostrongyloidea sister group to a clade including the Molineoidea and Heligmosomoidea. Within the Trichostrongyloidea, the Cooperiidae, Trichostrongylidae, and Haemonchidae were polytomous, the Haemonchinae were monophyletic, but the Ostertagiinae were paraphyletic. The sister-group relationships between Molineoidea and Heligmosomoidea were unsuspected from previous morphological analysis. No unequivocal morphological synapomorphy could be found for the grouping Molineoidea + Heligmosomoidea, but none was found which contradicted it. PMID- 11399152 TI - Interrelationships and evolution of the tapeworms (Platyhelminthes: Cestoda). AB - Interrelationships of the tapeworms (Platyhelminthes: Cestoda) were examined by use of small (SSU) and large (LSU) subunit ribosomal DNA sequences and morphological characters. Fifty new complete SSU sequences were added to 21 sequences previously determined, and 71 new LSU (D1-D3) sequences were determined for the complementary set of taxa representing each of the major lineages of cestodes as currently understood. New sequences were determined for three amphilinidean taxa, but were removed from both alignments due to their excessively high degree of divergence from other cestode sequences. A morphological character matrix coded for supraspecific taxa was constructed by the modification of matrices from recently published studies. Maximum-parsimony (MP) analyses were performed on the LSU, SSU, LSU+SSU, and morphological data partitions, and minimum-evolution (ME) analyses utilizing a general time reversible model of nucleotide substitution including estimates of among-site rate heterogeneity were performed on the molecular data partitions. Resulting topologies were rooted at the node separating the Gyrocotylidea from the Eucestoda. The LSU data were found to be more informative than the SSU data and were more consistent with inferences from morphology, although nodal support was generally weak for most basal nodes. One class of transitions was found to be saturated for comparisons between the most distantly related taxa (gyrocotylideans vs cyclophyllideans and tetrabothriideans). Differences in the topologies resulting from MP and ME analyses were not statistically significant. Nonstrobilate orders formed the basal lineages of trees resulting from analysis of LSU data and morphology. Difossate orders were basal to tetrafossate orders, the latter of which formed a strongly supported clade. A clade including the orders Cyclophyllidea, Nippotaeniidea, and Tetrabothriidea was supported by all data partitions and methods of analysis. Paraphyly of the orders Pseudophyllidea, Tetraphyllidea, and Trypanorhyncha was consistent among the molecular data partitions. Inferences are made regarding a monozoic (nonsegmented) origin of the Eucestoda as represented by the Caryophyllidea and for the evolution of the strobilate and acetabulate/tetrafossate conditions having evolved in a stepwise pattern. PMID- 11399153 TI - The molecular systematics and biogeography of the European cobitids based on mitochondrial DNA sequences. AB - Phylogenetic inference regarding the biogeography and evolution of the family Cobitidae depends in large part on the correct interpretation of transitions between the morphological states of secondary sexual characters (e.g., the scale of Canestrini or lamina circularis). Here, we use the complete mitochondrial ATP synthase 8 and 6 and cytochrome b genes to provide an independent assessment of systematics and biogeographic relationships of species in the genus Cobitis, including geographic and subgeneric sampling of species with Canestrini's scale present, duplicated, or absent. The mtDNA-based phylogeny for the genus Cobitis provides the first formal hypothesis for the group and permits a phylogenetic based assessment of the morphological transitions demonstrated by Canestrini's scale. Our data confirm the monophyly of the genus Cobitis and indicate that European Cobitis comprise six evolutionarily independent lineages. These lineages were defined by nucleotide synapomorphies permitting bootstrapped confidence estimates of 95% or greater and mtDNA genetic distances greater than 4.5% and correspond with moderate fidelity to the Cobitis groups defined by Bacescu (1962, Rev. Roum. Biol. 4, 435-448). The Caucasian lineage, C. cf. sibirica, represents the basal sister species of the genus Cobitis, supporting an eastern Asiatic origin of the European Cobitis: Cobitis sensu stricto, Acanestrinia, Bicanestrinia, Iberocobitis, and Cobitis calderoni. Phylogenetic relationships among Cobitis subgenera and species indicate that the ancestral condition of one scale of Canestrini was duplicated once at the origin of the Bicanestrinia lineage and has been independently lost by C. calderoni and C. elongata. The absence of the scale of Canestrini is the synapomorphy defining the subgenus Acanestrinia, but the mtDNA phylogeny indicates that Acanestrinia is not a natural group and places C. calderoni as the sister lineage to the subgenus Iberocobitis, a finding that is also geographically parsimonius. PMID- 11399154 TI - Parallel acceleration of evolutionary rates in symbiont genes underlying host nutrition. AB - The overproduction of essential amino acids by Buchnera aphidicola, the primary bacterial mutualist of aphids, is considered an adaptation for increased production of nutrients that are lacking in aphids' diet of plant sap. Given their shared role in host nutrition, amino acid biosynthetic genes of Buchnera are expected to experience parallel changes in selection that depend on host diet quality, growth rate, and population structure. This study evaluates the hypothesis of parallel selection across biosynthetic pathways by testing for correlated changes in evolutionary rates at biosynthetic genes of Buchnera. Previous studies show fast evolutionary rates at tryptophan biosynthetic genes among Buchnera associated with the aphid genus Uroleucon and suggest reduced purifying selection on symbiont nutritional functions in this aphid group. Here, we test for parallel rate acceleration at other amino acid biosynthetic genes of Buchnera-Uroleucon, including those for leucine (leuABC) and isoleucine/valine biosynthesis (ilvC). Ratios of nonsynonymous to synonymous substitutions (d(N)/d(S)) were estimated using codon-based maximum-likelihood methods that account for the extreme AT compositional bias of Buchnera sequences. A significant elevation in d(N)/d(S) at biosynthetic loci but not at two housekeeping genes sampled (dnaN and tuf) suggests reduced host-level selection on biosynthetic capabilities of Buchnera-Uroleucon. In addition, the discovery of trpEG pseudogenes in Buchnera-U. obscurum further supports reduced selection on amino acid biosynthesis. PMID- 11399155 TI - Molecular phylogeny of the major hylobatid divisions. AB - We describe DNA sequences for the mitochondrial control region and phenylalanine tRNA from the four extant gibbon subgenera. In contrast to earlier studies on gibbon phylogeny that used other parts of the mtDNA, the control region depicts the crested gibbons (Nomascus) as the most basal group of the Hylobatidae, followed by Symphalangus, with Bunopithecus and Hylobates as the last to diverge. Our data show that the molecular distances among the four gibbon subgenera are in the same range as those between Homo and Pan, or even higher. As a consequence of these findings, we propose to raise all four gibbon subgenera to genus rank. PMID- 11399157 TI - Alpha-Na6Pb3(PS4)4, a noncentrosymmetric thiophosphate with the novel, saucer shaped [Pb3(PS4)4]6- cluster, and its metastable, 3-dimensionally polymerized allotrope beta-Na6Pb3(PS4)4. PMID- 11399156 TI - Multiple hydrogen bond stabilization of a sandwich complex of sulfate between two macrocyclic tetraamides. PMID- 11399158 TI - Flexible redox-active binuclear macrocycles formed via the weak-link approach and novel hemilabile ligands with N,N,N',N'-tetramethyl-1,4-phenylenediamine units. PMID- 11399159 TI - Rhenium oxo complexes of a chelating diyne ligand. Synthesis and study of the kinetics of protonation. AB - A series of oxo complexes, Re(O)X(diyne) (X = I, Me, Et), have been prepared from 2,7-nonadiyne and Re(O)I(3)(PPh(3))(2). Addition of B(C(6)F(5))(3) to Re(O)I(2,7 nonadiyne) (5) results in coordination of the oxo ligand to the boron. The protonation of Re(O)(X)(2-butyne)(2) and Re(O)(X)(2,7-nonadiyne)(2) with a variety of acids has been examined. With 5 and HBF(4)/Et(2)O, the ultimate product was [Re(CH(3)CN)(3)(I)(2,7-nonadiyne)](2+) (7). The conversion of 5 to 7 changes the conformation of the diyne ligand from a "chair" to a "boat" and shifts its propargylic protons considerably downfield in the (1)H NMR. The kinetics of the protonation of Re(O)I(2,7-nonadiyne) (5) by CF(3)SO(3)H in CH(3)CN have been monitored by visible spectroscopy, in a stopped-flow apparatus, and by low temperature (1)H NMR. Two second-order rate constants, presumably successive protonations, were observed in the stopped-flow, k(1) = 11.9 M(-)(1) s(-)(1) and k(2) = 3.8 M(-)(1) s(-)(1). Low temperature (1)H NMR spectroscopy indicated that the resulting solution contained a mixture of two doubly protonated intermediates X and Y, each of which slowly formed the product 7 via an acid-independent process. PMID- 11399160 TI - Solid state and solution studies of lanthanide(III) complexes of cyclohexanetriols, models of the coordination sites found in sugars. AB - This report covers studies in trivalent lanthanide complexation by two simple cyclohexanetriols that are models of the two coordination sites found in sugars and derivatives. Several complexes of trivalent lanthanide ions with cis,cis 1,3,5-trihydroxycyclohexane (L(1)()) and cis,cis-1,2,3-trihydroxycyclohexane (L(2)()) have been characterized in the solid state, and some of them have been studied in organic solutions. With L(1)(), Ln(L)(2) complexes are obtained when crystallization is performed from acetonitrile solutions whatever the nature of the salt (nitrate or triflate) [Ln(L(1)())(2)(NO(3))(2)](NO(3)) (Ln = Pr, Nd); [Ln(L(1)())(2)(NO(3))H(2)O](NO(3))(2) (Ln = Eu, Ho, Yb); [Ln(L(1)())(2)(OTf)(2)(H(2)O)](OTf) (Ln = Nd, Eu). Lanthanum nitrate itself gives a mixed complex [La(L(1)())(2)(NO(3))(2)][LaL(1)()(NO(3))(4)] from acetonitrile solution while [La(L(1)())(2)(NO(3))(2)](NO(3)) is obtained using dimethoxyethane as reaction solvent and crystallization medium. With L(2)(), Ln(L)(2) complexes have also been crystallized from methanol solution [Ln(L(2)())(2)(NO(3))(2)]NO(3), (Ln = Pr, Nd, Eu). Single-crystal X-ray diffraction analyses are reported for these complexes. Complex formation in solution has been studied for several triflate salts (La, Pr, Nd, Eu, and Yb) with L(1 )()and L(2)(), respectively in acetonitrile and in methanol. In contrast to the solid state, both structures Ln(L) and Ln(L)(2) equilibrate in solution, as was demonstrated by low-temperature (1)H NMR and electrospray ionization mass spectrometry experiments. Competing experiments in complexing abilities of L(1)() and L(2)() with trivalent lanthanide cations have shown that only L(2)() exhibits a small selectivity (Nd > Pr > Yb > La > Eu) in methanol. PMID- 11399161 TI - Metal- and ligand-centered monoelectronic oxidation of mu nitrido[((tetraphenylporphyrinato)manganese)phthalocyaninatoiron)], [(TPP)Mn-N FePc]. X-ray crystal structure of the Fe(IV)-containing species [(THF)(TPP)Mn-N FePc(H(2)O)](I(5))02THF. AB - The reaction of mu nitrido[((tetraphenylporphyrinato)manganese)(phthalocyaninatoiron)], [(TPP)Mn-N FePc], with I(2) in THF develops with the formation of two different species, i.e., [(THF)(TPP)Mn-N-FePc(H(2)O)](I(5)).2THF (I) and [(TPP)Mn(IV)-N Fe(III)Pc](I(3)) (II). On the basis of single-crystal X-ray work and Mossbauer, EPR, Raman, and magnetic susceptibility data, I, found to be isostructural with the corresponding Fe-Fe complex, is shown to contain a low-spin triatomic Mn(IV)=N=Fe(IV) system (metal-centered oxidation). Data at hand for II (Mossbauer, EPR, Raman) show, instead, that oxidation takes place at one of the two macrocycles, very likely TPP (ligand-centered oxidation). The same cationic fragment present in I, and containing the Mn(IV)=N=Fe(IV) bond system, is also obtained when (TPP)Mn-N-FePc is allowed to react in THF with (phen)SbCl(6) (molar ratio 1:1). There are indications that the use of (phen)SbCl(6) in excess (2:1 molar ratio), in benzene, probably determines further oxidation with the formation of a species showing the combined presence of the Mn(IV)-Fe(IV) couple and of a pi-cation radical. PMID- 11399162 TI - Protonation and Zn(II) coordination by dipyridine-containing macrocycles with different molecular architecture. A case of pH-controlled metal jumping outside inside the macrocyclic cavity. AB - The synthesis of the macrocyclic ligand 4,4'-(2,5,8,11,14-pentaaza[15])-2,2' bipyridylophane (L3), which contains a pentaamine chain linking the 4,4' positions of a 2,2'-dipyridine moiety, is reported. Protonation and Zn(II) complexation by L3 and by macrocycle L2, containing the same pentaamine chain connecting the 6,6'-positions of 2,2'-dipyridine, were studied by means of potentiometric, UV-vis, and fluorescent emission measurements. While in L2 all the nitrogen donor atoms are convergent inside the macrocyclic cavity, in L3 the heteroaromatic nitrogen atoms are located outside. Both ligands form mono- and dinuclear Zn(II) complexes in aqueous solution. In the mononuclear Zn(II) complexes with L2, the metal is coordinated inside the macrocyclic cavity, bound to the heteroaromatic nitrogen donors and three amine groups of the aliphatic chain. As shown by the crystal structure of the [ZnL2](2+) complex, the two benzylic nitrogens are not coordinated and facile protonation of the complex takes place at slightly acidic pH values. Considering the mononuclear [ZnL3](2+) complex, the metal is encapsulated inside the cavity, not coordinated by the dipyridine unit. Protonation of the complex occurs on the aliphatic polyamine chain and gives rise to translocation of the metal outside the cavity, bound to the heteroaromatic nitrogens. PMID- 11399163 TI - A theoretical study of the structure of tricarbonatodioxouranate. AB - The results of a study on the ground states of tricarbonato complexes of dioxouranate using multiconfigurational second-order perturbation theory (CASSCF/CASPT2) are presented. The equilibrium geometries of the complexes corresponding to uranium in the formal oxidation states VI and V, [UO(2)(CO(3))(3)](4)(-) and [UO(2)(CO(3))(3)],(5)(-) have been fully optimized in D(3)(h)() symmetry at second-order perturbation theory (MBPT2) level of theory in the presence of an aqueous environment modeled by a reaction field Hamiltonian with a spherical cavity. The uranyl fragment has also been optimized at CASSCF/CASPT2, to obtain an estimate of the MBPT2 error. Finally, the effect of distorting the D(3)(h)() symmetry to C(3) has been investigated. This study shows that only minor geometrical rearrangements occur in the one-electron reduction of [UO(2)(CO(3))(3)](4)(-) to [UO(2)(CO(3))(3)],(5)(-) confirming the reversibility of this reduction. PMID- 11399164 TI - Heterobimetallic, cubane-like Mo(3)S(4)M' cluster cores containing the noble metals M' = Ru, Os, Rh, Ir. Unprecedented tri(mu-carbonyl) bridge between ruthenium atoms in [(eta(5)-Cp')(3)Mo(3)S(4)Ru)2(mu-CO)3]2+. AB - Reaction of the methylcyclopentadienyl (Cp') cluster compound [(eta(5) Cp')(3)Mo(3)S(4)][pts] (pts = p-toluenesulfonate) with noble metal alkene complexes resulted in the formation of four new heterobimetallic cubane-like Mo(3)S(4)M' cluster cores (M' = Ru, Os, Rh, Ir). Thus, reaction with [(1,5 cod)Ru(CO)(3)] or [(1,3-cod)Os(CO)(3)] (cod = cyclooctadiene) afforded [(eta(5) Cp')(3)Mo(3)S(4)M'(CO)(2)][pts] (M' = Ru: [1][pts]; M' = Os: [2][pts]). When [1][pts] was kept in CH(2)Cl(2)/pentane solution, partial loss of carbonyl ligands occurred and the carbonyl-bridged dicubane cluster [((eta(5) Cp')(3)Mo(3)S(4)Ru)(2)(mu-CO)(3)][pts](2) was isolated. An X-ray crystal structure revealed the presence of the hitherto unobserved Ru(mu-CO)(3)Ru structural element. The formation of cluster compounds containing Mo(3)S(4)Rh and Mo(3)S(4)Ir cores was achieved in boiling methanol by reacting [(eta(5) Cp')(3)Mo(3)S(4)][pts] with [M'Cl(cyclooctene)(2)](2) (M' = Rh, Ir) in the presence of PPh(3). In this way [(eta(5)-Cp')(3)Mo(3)S(4)M'Cl(PPh(3))][pts] (M' = Rh, Ir) could be isolated. An alternative route to the Mo(3)S(4)Rh cluster core was found in the reaction of [(eta(5)-Cp')(3)Mo(3)S(4)][pts] with [RhCl(1,5 cod)](2), which yielded [(eta(5)-Cp')(3)Mo(3)S(4)Rh(cod)][pts](2) ([7][pts](2)). Substitution of the cod ligand in [7][pts](2) by 1,3 bis(diphenylphosphanyl)propane (dppp) gave [(eta(5) Cp')(3)Mo(3)S(4)Rh(dppp)][pts](2). PMID- 11399165 TI - Magnetic anisotropy of a cyclic octanuclear Fe(III) cluster and magneto structural correlations in molecular ferric wheels. AB - The magnetic anisotropy of the cyclic octanuclear Fe(III) cluster [Cs subsetFe(8)[N(CH(2)CH(2)O)(3)](8)]Cl was investigated. Based on a spin Hamiltonian formalism and the consequent use of all symmetries, the magnetic anisotropy could be calculated exactly to first order, i.e., in the strong exchange limit. Experimentally, the magnetic anisotropy was investigated by magnetic susceptibility and high-field torque magnetometry of single crystals. The field and angle dependence of the torque at 1.7 K could be accurately reproduced by the calculations with one single parameter set, providing accurate results for the coupling constant and single-ion zero-field-splitting. These magnetic parameters are compared to those of several related hexanuclear ferric wheels and are discussed with respect to magneto-structural correlations for both coupling constant and single-ion anisotropy. PMID- 11399166 TI - Mechanism of sulfide oxidations by peroxymonocarbonate. AB - A detailed mechanism for the oxidation of aryl sulfides by peroxymonocarbonate ion in cosolvent/water media is described. Kinetic studies were performed to characterize the transition state, including a Hammett correlation and variation of solvent composition. The results are consistent with a charge-separated transition state relative to the reactants, with an increase of positive charge on the sulfur following nucleophilic attack of the sulfide at the electrophilic oxygen of peroxymonocarbonate. In addition, an average solvent isotope effect of 1.5 +/- 0.2 for most aryl sulfide oxidations is consistent with proton transfer in the transition state of the rate-determining step. Activation parameters for oxidation of ethyl phenyl sulfide in tert-butyl alcohol/water are reported. From the pH dependence of oxidation rates and (13)C NMR equilibrium experiments, the estimated pK(a) of peroxymonocarbonate was found to be approximately 10.6. PMID- 11399167 TI - X-ray crystal structures of alpha KrF(2),[KrF][MF(6)](M=As,Sb,Bi),[Kr(2)F(3)][SbF(6).KrF(2), [Ke(2)F(3)2[SbF(6)]2.KrF(2), and [Kr(2)F(3)][AsF(6)].[KrF][AsF(6)]; synthesis and characterization of [Kr(2)F(3)][PF(6).nKrF(2); and theoretical studies of KrF(2), KrF+, Kr(2)F(3)+, and the [KrF][MF(6)](M=P,As,Sb,Bi) ion pairs. AB - The crystal structures of alpha-KrF(2) and salts containing the KrF(+) and Kr(2)F(3)(+) cations have been investigated for the first time using low temperature single-crystal X-ray diffraction. The low-temperature alpha-phase of KrF(2) crystallizes in the tetragonal space group I4/mmm with a = 4.1790(6) A, c = 6.489(1) A, Z = 2, V = 113.32(3) A(3), R(1) = 0.0231, and wR(2) = 0.0534 at 125 degrees C. The [KrF][MF(6)] (M = As, Sb, Bi) salts are isomorphous and isostructural and crystallize in the monoclinic space group P2(1)/c with Z = 4. The unit cell parameters are as follows: beta-[KrF][AsF(6)], a = 5.1753(2) A, b = 10.2019(7) A, c = 10.5763(8) A, beta = 95.298(2) degrees, V = 556.02(6) A(3), R(1) = 0.0265, and wR(2) = 0.0652 at -120 degrees C; [KrF][SbF(6)], a = 5.2922(6) A, b = 10.444(1) A, c = 10.796(1) A, beta = 94.693(4) degrees, V = 594.73(1) A(3), R(1) = 0.0266, wR(2) = 0.0526 at -113 degrees C; [KrF][BiF(6)], a = 5.336(1) A, b = 10.513(2) A, c = 11.046(2) A, beta = 94.79(3) degrees, V = 617.6(2) A(3), R(1) = 0.0344, and wR(2) = 0.0912 at -130 degrees C. The Kr(2)F(3)(+) cation was investigated in [Kr(2)F(3)][SbF(6)].KrF(2), [Kr(2)F(3)](2)[SbF(6)](2).KrF(2), and [Kr(2)F(3)][AsF(6)].[KrF][AsF(6)]. [Kr(2)F(3)](2)[SbF(6)](2).KrF(2) crystallizes in the monoclinic P2(1)/c space group with Z = 4 and a = 8.042(2) A, b = 30.815(6) A, c = 8.137(2) A, beta = 111.945(2) degrees, V = 1870.1(7) A(3), R(1) = 0.0376, and wR(2) = 0.0742 at -125 degrees C. [Kr(2)F(3)][SbF(6)].KrF(2) crystallizes in the triclinic P1 space group with Z = 2 and a = 8.032(3) A, b = 8.559(4) A, c = 8.948(4) A, alpha = 69.659(9) degrees, beta = 63.75(1) degrees, gamma = 82.60(1) degrees, V = 517.1(4) A(3), R(1) = 0.0402, and wR(2) = 0.1039 at -113 degrees C. [Kr(2)F(3)][AsF(6)].[KrF][AsF(6)] crystallizes in the monoclinic space group P2(1)/c with Z = 4 and a = 6.247(1) A, b = 24.705(4) A, c = 8.8616(6) A, beta = 90.304(6) degrees, V = 1367.6(3) A(3), R(1) = 0.0471 and wR(2) = 0.0958 at -120 degrees C. The terminal Kr-F bond lengths of KrF(+) and Kr(2)F(3)(+) are very similar, exhibiting no crystallographically significant variation in the structures investigated (range, 1.765(3)-1.774(6) A and 1.780(7)-1.805(5) A, respectively). The Kr-F bridge bond lengths are significantly longer, with values ranging from 2.089(6) to 2.140(3) A in the KrF(+) salts and from 2.027(5) to 2.065(4) A in the Kr(2)F(3)(+) salts. The Kr-F bond lengths of KrF(2) in [Kr(2)F(3)][SbF(6)].KrF(2) and [Kr(2)F(3)](2)[SbF(6)](2).KrF(2) range from 1.868(4) to 1.888(4) A and are similar to those observed in alpha-KrF(2) (1.894(5) A). The synthesis and Raman spectrum of the new salt, [Kr(2)F(3)][PF(6)].nKrF(2), are also reported. Electron structure calculations at the Hartree-Fock and local density-functional theory levels were used to calculate the gas-phase geometries, charges, Mayer bond orders, and Mayer valencies of KrF(+), KrF(2), Kr(2)F(3)(+), and the ion pairs, [KrF][MF(6)] (M = P, As, Sb, Bi), and to assign their experimental vibrational frequencies. PMID- 11399168 TI - Ability of a Au(III)-N unit to bond two aurophilically interacting gold(I) centers. AB - The monohapto neutral 2-(diphenylphosphino)aniline (PNH(2)) complexes [Au(C(6)F(5))(2)X(PNH(2))] (X = C(6)F(5) (1), Cl (2)) have been obtained from [Au(C(6)F(5))(3)(tht)] or [Au(C(6)F(5))(2)(micro-Cl)](2) and PNH(2), and the cationic [Au(C(6)F(5))(2)(PNH(2))]ClO(4) (3) has been similarly prepared from [Au(C(6)F(5))(2)(OEt(2))(2)]ClO(4) and PNH(2) or from 2 and AgClO(4). The neutral amido complex [Au(C(6)F(5))(2)(PNH)] (4) can be obtained by deprotonation of 3 with PPN(acac) (acac = acetylacetonate) or by treatment of the chloro complex 2 with Tl(acac). It reacts with [Ag(OClO(3))(PPh(3))] or [Au(OClO(3))(PPh(3))] to give the dinuclear species [Au(C(6)F(5))(2)[PNH(MPPh(3))]]ClO(4) (M = Ag (5), Au (6)). The latter can also be obtained by reaction of equimolar amounts of 3 and [Au(acac)(PPh(3))]; when the molar ratio of the same reagents is 1:2, the trinuclear cationic complex [Au(C(6)F(5))(2)[PN(AuPPh(3))(2)]]ClO(4) (7) is obtained. The crystal structures of complexes 2-4 and 7 have been established by X-ray crystallography; the last-mentioned displays an unusual Au(I)-Au(III) interaction. PMID- 11399169 TI - Strong cooperativeness in the mononuclear iron(II) derivative exhibiting an abrupt spin transition above 400 K. AB - The spin crossover system, [Fe(bzimpy)(2)](ClO(4))(2).0.25H(2)O, was reinvestigated above room temperature (bzimpy = 2,6-bis(benzimidazol-2 yl)pyridine). The system exhibits an abrupt low-spin to high-spin transition at T(c) = 403 K. Liberation of a fractional amount of water does not affect the spin crossover: the system is perfectly reversible with a hysteresis width of DeltaT = 12 K. The existence of the hysteresis at such high temperature determines that the lowest limit of the solid-state cooperativity parameter is J/k > 403 K despite long iron(II) separations (10 A). The high cooperativeness has been assigned to a perfect pi-stacking of the benzimidazole rings in the crystal lattice at a distance as short as 3.6 A. Variable-temperature IR data and the heat capacity measurements match well the magnetic data. The thermodynamic properties are DeltaH = 17 kJ mol(-)(1), DeltaS = 43 J K(-)(1) mol(-)(1), so that the entropy of the spin transition shows a considerable contribution from the molecular vibrations. A theoretical model has been applied in fitting the magnetic data along the whole hysteresis path. A statistical distribution of the cooperativity parameter led to the feature that angled walls of the hysteresis loop are well reproduced. PMID- 11399170 TI - Preparation and characterization of ruthenium(II) monophosphaferrocene complexes. Reactivity, dynamic solution behavior, and X-ray structure of [RuH(2)(eta(2) H(2))(PCy(3))2(2-phenyl-3,4-dimethylphosphaferrocene)]. AB - The bis(dihydrogen) complex RuH(2)(H(2))(2)(PCy(3))(2) (1) reacts with 2-phenyl 3,4-dimethylphosphaferrocene (L(1)) to give RuH(2)(H(2))(PCy(3))(2)(L(1)) (2). This dihydride-dihydrogen complex has been characterized by X-ray crystallography and variable-temperature (1)H and (31)P NMR spectroscopy. The exchange between the dihydrogen ligand and the two hydrides is characterized by a DeltaG() of 46.2 kJ/mol at 263 K. H/D exchange is readily observed when heating a C(7)D(8) solution of 2 (J(H-D) = 30 Hz). The H(2) ligand in 2 can be displaced by ethylene or carbon monoxide leading to the corresponding ethylene or carbonyl complexes. The reaction of 1 with 2 equiv of 3,4-dimethylphosphaferrocene (L(2)) yields the dihydride complex RuH(2)(PCy(3))(2)(L(2))(2) (5). PMID- 11399171 TI - Preparation and conformational properties of the perfluoro diether CF(3)OCF(2)OCF(2)C(O)F, a model molecule to study properties of perfluoro polyethers. AB - The compound CF(3)OCF(2)OCF(2)C(O)F was prepared by oxidation of hexafluoropropene with molecular oxygen in the gas-phase using CF(3)OF as initiator. (13)C NMR, FTIR, Raman, UV-vis, and mass spectra were obtained and interpreted. The theoretical structure studies were performed by the calculation of the potential energy surfaces, using the results obtained for a smaller related molecule, CF(3)OCF(2)C(O)F, as a starting point. A high degree of conformational flexibility of this compound is evidenced by the values of several conformations, varying within the range of 1 kcal/mol. Theoretical calculations predict chain conformations as the most stable molecular forms, as expected from the presence of the anomeric effect. The experimental fundamental vibrational modes are compared with those obtained theoretically, using ab initio and density functional theory methods, HF/6-31+G and B3LYP/6-31+G, respectively. The density of the compound at ambient temperature (delta = 1.7(1) g/mL), its melting point (mp = -140(5) degrees C), its boiling point (bp = 14.5 (1) degrees C), and the relation between its vapor pressure and the absolute temperature (ln P = 13.699 - 2023.4/T) were also determined. PMID- 11399172 TI - Structure, stability, and interconversion barriers of the rotamers of cis [Pt(II)Cl(2)(quinoline)2] and cis-[Pt(II)Cl(2)(3-bromoquinoline)(quinoline)] from X-ray crystallography, NMR spectroscopy and molecular mechanics evidence. AB - Reported are the preparations of cis-[PtCl(2)(quinoline)(2)] and cis-[PtCl(2)(3 bromoquinoline)(quinoline)] and an investigation of the stabilities and interconversion of the rotamer forms of these complexes. Both head-to-head (HTH) and head-to-tail (HTT) rotamer forms are found in the crystal structure of cis [PtCl(2)(quinoline)(2)]. The NOESY NMR spectrum of cis-[PtCl(2)(quinoline)(2)] in dmf-d(7) at 300 K is consistent with conformational exchange brought about by rotation about the Pt-N(quinoline) bonds. H.H nonbonded distances between H atoms of the two different quinoline ligands were determined from NOESY data, and these distances are in accord with those observed in the crystal structure and derived from molecular mechanics models. cis-[PtCl(2)(3-bromoquinoline)(quinoline)] was prepared to alleviate the symmetry-imposed absence of inter-ring H2/H2 and H8/H8 NOESY cross-peaks for cis-[PtCl(2)(quinoline)(2)]. Molecular mechanics calculations on the complexes show the HTT rotamers to be 1-2 kJ mol(-)(1) more stable than the HTH forms, consistent with the (1)H spectra where the intensities of resonances for the two forms are approximately equal. Variable-temperature (1)H NMR spectra of cis-[PtCl(2)(quinoline)(2)] in dmf-d(7) indicate a rotational energy barrier of 82 +/- 4 kJ mol(-)(1). Variable-temperature (1)H NMR spectra indicate that the Br substituent on the quinoline ring does not affect the energy barrier to interconversion between the HTT and HTH forms (79 +/- 5 kJ mol(-)(1)). The steric contribution to the rotation barrier was calculated using molecular mechanics calculations and was found to be approximately 40 kJ mol(-)(1), pointing to a possible need for an electronic component to be included in future models. PMID- 11399173 TI - Mechanism of protonation of [Pt(3)(mu-PBu(t)(2))3(H)(CO)2], yielding the hydride bridged [Pt(3)(mu-PBu(t)(2))2(mu-H)(PBu(t)(2)H)(CO)2]OTf (Tf=CF(3)SO(2)), and the spectroscopic and theoretical characterization of a kinetic intermediate. AB - The reaction of the Pt(I)Pt(I)Pt(II) triangulo cluster Pt(3)(micro PBu(t)()(2))(3)(H)(CO)(2) (1) with TfOH (Tf = CF(3)SO(2)) affords the hydride bridged cationic derivative [Pt(3)(mu-PBu(t)()(2))(2)(mu H)(PBu(t)()(2)H)(CO)(2)]OTf (2). With TfOD the reaction gives selectively [Pt(3)(mu-PBu(t)(2))(2)(mu-D)(PBu(t)(2)H)(CO)(2)]OTf (2-D(1)), implying that the proton is transferred to a metal center while a P-H bond is formed by the reductive coupling of one of the bridging phosphides and the terminal hydride ligand of the reagent. The reaction proceeds through the formation of a thermally unstable kinetic intermediate which was characterized at low temperatures, and was suggested to be the CO-hydrogen-bonded (or protonated) [Pt(3)(mu PBu(t)(2))(3)(H)(CO)(2)].HOTf (3). An ab initio theoretical study predicts a hydrogen-bonded complex or a proton-transfer tight ion pair as a possible candidate for the structure of the kinetic intermediate. PMID- 11399174 TI - Oxidation state dependence of the geometry, electronic structure, and magnetic coupling in mixed oxo- and carboxylato-bridged manganese dimers. AB - Approximate density functional theory has been used to investigate changes in the geometry and electronic structure of the mixed oxo- and carboxylato-bridged dimers [Mn(2)(mu-O)(2)(O(2)CH)(NH(3))(6)](n+)and [Mn(2)(mu O)(O(2)CH)(2)(NH(3))(6)](n+)in the Mn(IV)Mn(IV), Mn(III)Mn(IV), and Mn(III)Mn(III) oxidation states. The magnetic coupling in the dimer is profoundly affected by changes in both the bridging ligands and Mn oxidation state. In particular, change in the bridging structure has a dramatic effect on the nature of the Jahn-Teller distortion observed for the Mn(III) centers in the III/III and III/IV dimers. The principal magnetic interactions in [Mn(2)(mu O)(2)(O(2)CH)(NH(3))(6)](n+)() involve the J(xz/xz)and J(yz/yz) pathways but due to the tilt of the Mn(2)O(2) core, they are less efficient than in the planar di mu-oxo structure and, consequently, the calculated exchange coupling constants are generally smaller. In both the III/III and III/IV dimers, the Mn(III) centers are high-spin, and the Jahn-Teller effect gives rise to axially elongated Mn(III) geometries with the distortion axis along the Mn-O(c) bonds. In the III/IV dimer, the tilt of the Mn(2)O(2) core enhances the crossed exchange J(x)()()2( )(y)()()2(/)(z)()()2 pathway relative to the planar di-mu-oxo counterpart, leading to significant delocalization of the odd electron. Since this delocalization pathway partially converts the Mn(IV) ion into low-spin Mn(III), the magnetic exchange in the ground state can be considered to arise from two interacting spin ladders, one is the result of coupling between Mn(IV) (S = 3/2) and high-spin Mn(III) (S = 2), the other is the result of coupling between Mn(IV) (S = 3/2) and low-spin Mn(III) (S = 1). In [Mn(2)(mu O)(O(2)CH)(2)(NH(3))(6)](n+)(), both the III/III dimer and the lowest energy structure for the III/IV dimer involve high-spin Mn(III), but the Jahn-Teller axis is now orientated along the Mn-oxo bond, giving rise to axially compressed Mn(III) geometries with long Mn-O(c) equatorial bonds. In the IV/IV dimer, the ferromagnetic crossed exchange J(yz)()(/)(z)()()2 pathway partially cancels J(yz/yz) and, as a consequence, the antiferromagnetic J(xz/xz) pathway dominates the magnetic coupling. In the III/III dimer, the J(yz/yz) pathway is minimized due to the smaller Mn-O-Mn angle, and since the ferromagnetic J(yz)()(/)(z)()()2 pathway largely negates J(xz/xz), relatively weak overall antiferromagnetic coupling results. In the III/IV dimer, the structures involving high-spin and low spin Mn(III) are almost degenerate. In the high-spin case, the odd electron is localized on the Mn(III) center, and the resulting antiferromagnetic coupling is similar to that found for the IV/IV dimer. In the alternative low-spin structure, the odd electron is significantly delocalized due to the crossed J(yz)()(/)(z)()()2 pathway, and cancellation between ferromagnetic and antiferromagnetic pathways leads to overall weak magnetic coupling. The delocalization partially converts the Mn(IV) ion into high-spin Mn(III), and consequently, the spin ladders arising from coupling of Mn(IV) (S = 3/2) with high-spin (S = 2) and low-spin (S = 1) Mn(III) are configurationally mixed. Thus, in principle, the ground-state magnetic coupling in the mixed-valence dimer will involve contributions from three spin-ladders, two associated with the delocalized low-spin structure and the third arising from the localized high-spin structure. PMID- 11399175 TI - Synthesis, characterization, and magnetic properties of self-assembled compounds based on discrete homotrinuclear complexes of Cu(II). AB - Three new supramolecular entities of Cu(II) were synthesized and characterized: [(Cu(H(2)O)(tmen))(2)(mu-Cu(H(2)O)(opba))](2)[(ClO(4))(2)](2).2H(2)O (1), [(Cu(H(2)O)(tmen))(2) (mu-Cu(H(2)O) (Me(2)pba))](2)[(ClO(4))(2)](2) (2), and [(Cu(H(2)O)(tmen))(Cu(tmen))(mu-Cu(OHpba))](n)() ((ClO(4))(2))(n)().nH(2)O (3), where opba = o-phenylenbis(oxamato), Me(2)pba = 2,2-dimethyl-1,3 propylenbis(oxamato), OHpba = 2-hydroxy-1,3-propylenbis(oxamato), and tmen = N,N,N'N'-tetramethylethylenediamine. The crystal structures of 1, 2, and 3 were solved. Complex 1 crystallizes in the monoclinic system, space group C2/c with a = 20.572(4) A, b = 17.279(6) A, c = 22.023(19) A, beta = 103.13(4) degrees, and Z = 8. Complex 2 crystallizes in the monoclinic system, space group P2(1)/c, with a = 16.7555(7) A, b = 13.5173(5) A, c = 17.1240(7) A, beta = 104.9840(10) degrees, and Z = 4. Complex 3 crystallizes in the orthorhombic system, space group Pca2(1) with a = 21.2859(4) A, b = 12.8286(10) A, c = 12.6456(2) A, and Z = 4. The three complexes are very similar in structure: a trinuclear Cu(II) complex with the two terminal Cu(II) ions blocked by N,N,N',N'-tetramethylethylenediamine, but with a different environment in the Cu(II) central ion. In the case of complex 1, two of these trinuclear entities are packed with a short distance between the central Cu(II) ions of two separate entities forming a hexanuclear-type compound. In the case of 2, two of these trinuclear entities are linked by a hydrogen bond between a water molecule of one terminal Cu(II) and one oxygen atom of the oxamato ligand of the neighboring entity, also forming a hexanuclear complex. In the case of complex 3, the intermolecular linkages give a one-dimensional system where the OH groups of the OHpba entities are linked to the terminal Cu(II) of the neighboring entities. The magnetic properties of the three complexes were studied by susceptibility measurements vs temperature. For complex 1, an intramolecular J value of -312.1 cm(-)(1) and a contact dipolar interaction of -0.44K were found. For complex 2 and 3 the fit was made by the irreducible tensor operator formalism (ITO). The values obtained were as follows: J(1) = -333.9 cm(-)(1) and J(2) = 0.67 cm(-)(1) for 2 and J(1) = -335.9 cm(-)(1) and J(2) = 3.5 cm(-)(1) for 3. PMID- 11399176 TI - Synthesis and properties of mercury bis(trifluoromethyl)phosphanides and their phosphane complexes. AB - The thermally unstable compounds Hg(CN)P(CF(3))(2) and Hg[P(CF(3))(2)](2) were obtained by reactions of mercury cyanide and bis(trifluoromethyl)phosphane in solution and characterized by multinuclear NMR spectroscopy. An increase in thermal stability is observed when the products form 18 valence electron complexes. The compounds [Hg(P(CF(3))(2))(2)(dppe)] (dppe = 1,2 bis(diphenylphosphanyl)ethane) and [Hg(P(CF(3))(2))(2)(Me(3)P)(2)] have been isolated in almost quantitative yield by reacting [Hg(CN)(2)(dppe)] or [Hg(CN)(2)(Me(3)P)(2)] with HP(CF(3))(2). [Hg(P(CF(3))(2))(2)(dppe)] crystallizes in the triclinic space group P1. The mercury atom is coordinated in a distorted tetrahedral fashion. The Hg-P(CF(3))(2) bonds, ca. 250 pm, are significantly longer than those of the mercury bis(phosphanides) Hg(PR(2))(2) with R = t-Bu, 245 pm, or SiMe(3), 241 pm. These easily accessible compounds [Hg(P(CF(3))(2))(2)(dppe)] and [Hg(P(CF(3))(2))(2)(Me(3)P)(2)] act as nucleophilic bis(trifluoromethyl)phosphane group transfer reagents. PMID- 11399177 TI - Copper(II) complexes with 4-amino-N-[4,6-dimethyl-2 pyrimidinyl]benzenesulfonamide. Synthesis, crystal structure, magnetic properties, EPR, and theoretical studies of a novel mixed mu-carboxylato, NCN bridged dinuclear copper compound. AB - New Cu(II) complexes of sulfamethazine (4-amino-N-[4,6-dimethyl-2 pyrimidinyl]benzenesulfonamide, HL) [Cu(2)(CH(3)COO)(2)(L)(2)].2dmf (1) and ([Cu(L)(2)].2H(2)O)(infinity) (2) were prepared and structurally characterized. Compound 1 crystallizes in the monoclinic system, space group P2(1)/n, with a = 8.9486(9) A, b = 15.0956(12) A, c = 16.542(3) A, beta = 105.584(15) degrees, and Z = 2. Compound 2 crystallizes in the monoclinic system, space group P2(1)/c, with a = 13.8097(8) A, b = 14.5765(4) A, c = 13.7853(15) A, beta = 96.033(9) degrees, and Z = 1. In compound 1 two copper ions are linked by two syn-syn acetates and two nonlinear NCN bridging groups pertaining to the deprotonated sulfamethazine ligands. Each copper center presents a nearly square planar geometry. Magnetic susceptibility data for 1 show a strong antiferromagnetic coupling with 2J = -216.7 cm(-)(1). The EPR spectra at the X- and Q-band frequencies present the signals corresponding to the dinuclear entity, being the zero-field splitting parameter, D = 0.265 cm(-)(1). The antiferromagnetic exchange coupling is discussed using DFT calculations on some model compounds with NCN bridging ligands and also on model structures with mixed mu-acetato and NCN bridges. The copper in the polymeric compound 2 is five coordinate. The CuN(5) chromophore has a highly distorted square pyramidal geometry with small axial N-Cu-N angles of 65.53(14) and 59.90(13) degrees. In the structure a sulfamethazinate anion binds to one copper through the sulfonamido and pyrimidine N atoms and to an adjacent copper via the amino N atom. PMID- 11399178 TI - 1,2-Bis(2-pyridylethynyl)benzene, a novel trans-chelating bipyridyl ligand. structural characterization of the complexes with silver(I) triflate and palladium(II) chloride. AB - The design, synthesis, and complexation characteristics of the bipyridyl ligand 1,2-bis-(2-pyridylethynyl)benzene are described. The X-ray crystallographic characterization of the 1:1 complexes of 1,2-bis(2-pyridylethynyl)benzene with silver(I) triflate and palladium(II) chloride are described. In the X-ray crystal structure of the silver(I) triflate complex the ligand is essentially planar with negligible distortion compatible with a good fit of the cation in the "cavity" between the pyridine N atoms. Indeed the silver center is almost linear with the N(1)-Ag(1)-N(2) angle of 177.02(10) degrees. The ligand is also essentially planar in the palladium(II) chloride complex with square planar coordination about the palladium with the N(1)-Pd(1)-N(2), Cl(2)-Pd(1)-Cl(2), and N(1)-Pd(1) Cl(2) angles at 179.53(7), 177.17(2), and 90.52(5) degrees, respectively. PMID- 11399179 TI - Electronic structure of 3d[M(H2O)6](3+) ions from Sc(III) to Fe(III): a quantum mechanical study based on DFT computations and natural bond orbital analyses. AB - The metal-donor atom bonding along the series of 3d[M(H2O)6](3+) ions from Sc(3+) to Fe(3+) has been investigated by density-functional calculations combined with natural localized bond orbital analyses. The M-OH(2) bonds were considered as donor-acceptor bonds, and the contributions coming from the metal ion's 3d sigma , 3d pi-, and 4s sigma-interactions were treated individually. In this way, the total amount of charge transferred from the water oxygen-donor atoms toward the appropriate metal orbitals could be analyzed in a straightforward manner. One result obtained along these lines is that the overall extent of ligand-to-metal charge transfer shows a strong correlation to the hydration enthalpies of the aqua metal ions. If the contributions to the total ligand-to-metal ion charge transfer are divided into sigma- and pi-contributions, it turns out that Cr(3+) is the best sigma-acceptor, but its pi-accepting abilities are the weakest along the series. Fe(3+) is found to be the best pi-acceptor among the 3d hexaaqua ions studied. Its aptitude to accept sigma-electron density is the second weakest along the series and only slightly higher than that of Sc(3+) (the least sigma acceptor of all ions) because of the larger involvement of the Fe(3+) 4s orbital in sigma-bonding. The strengths of the three types of bonding interactions have been correlated with the electron affinities of the different metal orbitals. Deviations from the regular trends of electron affinities along the series were found for those [M(H2O)6](3+) ions that are subject to Jahn-Teller distortions. In these cases (d(1) = [Ti(H2O)6](3+), d(2) = [V(H2O)6](3+), and d(4) = [Mn(H2O)6](3+)), ligand-to-metal charge transfer is prevented to go into those metal orbitals that contain unpaired d electrons. A lowering of the complex symmetry is observed and coupled with the following variations: The Ti(3+)- and V(3+)-hexaaqua ions switch from T(h)() to C(i)() symmetry while the Mn(3+) hexaaqua ion moves to D(2)(h)() symmetry. The loss of orbital overlap leading to a diminished ligand-to-metal charge transfer toward the single occupied metal orbitals is compensated by amplified bonding interactions of the ligand orbitals with the unoccupied metal orbitals to some extent. PMID- 11399180 TI - Preparation and characterization of bis(trifluoromethyl)phosphanide salts of outstanding stability. AB - The stable compounds [NEt(4)][P(CF(3))(2)] and [18-crown-6-K][P(CF(3))(2)] were synthesized in quantitative yields by treatment of HP(CF(3))(2) with ionic cyanides at low temperature. These novel salts were characterized by multinuclear NMR spectroscopy, elemental analysis, and vibrational spectroscopy. Excellent agreement of experimental and theoretical vibrational frequencies, calculated at the B3PW91 level of theory, clearly confirms the saltlike character of these compounds. Due to their ionic nature, these salts are excellent nucleophilic reagents for the transfer of P(CF(3))(2) groups, suitable for the synthesis of chiral bidentate bis(trifluoromethyl)phosphine containing compounds. PMID- 11399181 TI - Three conformational polymorphs of di-mu-chlorotetrakis(1 methylboratabenzene)diyttrium: synthesis, x-ray structures, quantum chemical calculations, and lattice energy minimizations. AB - The reaction of yttrium trichloride with lithium 1-methylboratabenzene (1/2) in toluene (110 degrees C, 3 days) afforded the donor-free dinuclear sandwich complex [(C(5)H(5)BMe)(2)Y(mu-Cl)](2) (1) in 85% yield as pale-yellow crystals. By means of single crystal and powder diffraction methods, three conformational polymorphs, alpha-1 [P2(1)/n (No. 14), monoclinic, a = 6.6124(8) A, b = 14.352(9) A, c = 14.120(1) A, beta = 95.57(1) degrees, V = 1333.7(9) A(3), Z = 2], beta-1 [P2(1)/a (No. 14), monoclinic, a = 8.542(2) A, b = 13.712(6) A, c = 11.76(1) A, beta = 102.60(4) degrees, V = 1344.5(13) A(3), Z = 2], and gamma-1 [Pbca (No. 61), orthorhombic, a = 20.091(5) A, b = 13.527(3) A, c = 9.976(2) A, V = 2711.2(11) A(3), Z = 4], were characterized in the solid state of 1. The molecules in the three phases vary remarkably in the rotational position of boratabenzene ligands with differences of 91.1, 133.1, and 24.9 degrees between each pair. DFT calculations at the B3LYP/LanL2DZ level reveal that the three molecular structures observed in the solid state correspond closely to three minima on the gas-phase potential energy surface. The beta conformation is 2.8 and 7.2 kJ/mol more stable than the alpha and gamma conformations, respectively. Lattice energy minimizations predict that the alpha-1 phase is about 5.5 and 18.7 kJ mol(-)(1) more stable than the beta-1 and gamma-1 modifications, in agreement with the packing coefficients and the molecular volumes of the three crystal structures. While the alpha-1 and beta-1 modifications have comparable total energies, the gamma-1 form is less stable. The total energy differences among the polymorphs are greater than generally expected. PMID- 11399182 TI - Cobalt(III) complexes of [3(5)] adamanzane, 1,5,9,13 tetraazabicyclo[7.7.3]nonadecane. Report of an inert, chelate hydrogen carbonate ion. AB - Three cobalt(III) complexes of the macrocyclic tetraamine [3(5)]adamanzane (1,5,9,13-tetraazabicyclo[7.7.3]nonadecane) were isolated as salts. The X-ray crystal structures were solved for the compounds [Co([3(5)]adz)(CO(3))]AsF(6) (1b), [Co([3(5)]adz)(HCO(3))]ZnBr(4).H(2)O (2a), and [Co([3(5)]adz)(SO(4))]AsF(6).H(2)O (3a). The coordination geometry around the cobalt(III) ion is a distorted octahedron with the inorganic ligands at cis positions. Complex 2 is the second example of a cobalt(III) complex for which the X-ray structure shows a chelate binding mode of the hydrogen carbonate entity. The pK(a) value of the [Co([3(5)]adz)(HCO(3))](2+) ion (2) was determined spectrophotometrically to be 0.27 (25 degrees C, I = 5.0 M). The protonation appears to occur at the noncoordinated carbonyl oxygen atom of the carbonate group, with hydrogen bonding to the crystal water molecule. Evidence is presented for this oxygen atom as the site of protonation in solution as well. In 5.0 M CF(3)SO(3)H a slow reaction of the carbonato complex, quantitatively yielding the [Co([3(5)]adz)(H(2)O)(2)](3+) ion, was observed. k(obs) = 7.9(1) x 10(-)(6) s( )(1) at 25 degrees C. PMID- 11399183 TI - Synthesis and characterization of CdIn(2)S(4) nanorods by converting CdS nanorods via the hydrothermal route. AB - The ternary semiconductor CdIn(2)S(4) nanorods were synthesized by a method based on CdS nanorods via the hydrothermal route, in which CdS nanorods were converted by reaction with InCl(3) and thiourea in aqueous solution. Transmission electron microscopy (TEM) images revealed that the typical sizes of the CdIn(2)S(4) nanorods were 10-30 nm in diameter and 200-1000 nm in length. X-ray photoelectron spectra (XPS) analysis of the surface stoichiometry (CdIn(2.03)S(4.15)) and room temperature Raman spectrum (RS) were recorded. The influences of reaction temperature, time, and sulfur sources on the formation for CdIn(2)S(4) nanorods were investigated. A possible formation mechanism of the CdIn(2)S(4) nanorods was also proposed. PMID- 11399184 TI - Oxamato-bridged trinuclear Ni(II)Cu(II)Ni(II) complexes with irregular spin state structures and a binuclear Ni(II)Cu(II) complex with an unusual supramolecular structure: crystal structure and magnetic properties. AB - Four oxamato-bridged heterotrinuclear Ni(II)Cu(II)Ni(II) complexes of formula ([Ni(bispictn)](2)Cu(pba))(ClO(4))(2).2.5H(2)O (1), ([Ni(bispictn)](2)Cu(pbaOH))(ClO(4))(2).H(2)O (2), ([Ni(cth)](2)Cu(pba))(ClO(4))(2) (3), and ([Ni(cth)](2)Cu(opba))(ClO(4))(2).H(2)O (4) and a binuclear Ni(II)Cu(II) complex of formula [Cu(opba)Ni(cth)].CH(3)OH (5) have been synthesized and characterized by means of elemental analysis, IR, ESR, and electronic spectra, where pba = 1,3-propylenebis(oxamato), pbaOH = 2-hydroxyl 1,3-propylenebis(oxamato), opba = o-phenylenebis(oxamato), bispictn = N,N'-bis(2 pyridylmethyl)-1,3-propanediamine, and cth = rac-5,7,7,12,14,14-hexamethyl 1,4,8,11-tetraazacyclotetradecane. The crystal structures of 1, 3, and 5 have been determined. The structures of complexes 1 and 3 consist of trinuclear cations and perchlorate anions, and that of 5 consists of neutral binuclear molecules which are connected by hydrogen bonds and pi-pi interactions to produce a unique supramolecular "double" sheet. In the three complexes, the copper atom in a square-planar or axially elongated octahedral environment and the nickel atom in a distorted octahedral environment are bridged by the oxamato groups, with Cu.Ni separations between 5.29 and 5.33 A. The magnetic properties of all five complexes have been investigated. The chi(M)T versus T plots for 1-4 exhibit the minimum characteristic of antiferromagnetically coupled NiCuNi species with an irregular spin state structure and a spin-quartet ground state. The chi(M)T versus T plot for 5 is typical of an antiferromagnetically coupled NiCu pair with a spin-doublet ground state. The Ni(II)-Cu(II) isotropic interaction parameters for the five complexes were evaluated and are between 102 and 108 cm(-)(1) (H = JS(Cu).S(Ni)). PMID- 11399185 TI - Syntheses, structures, and stabilities of [PPh(4)][WS(3)(SR)](R=(i)Bu,(i)Pr,(i)Bu, benzyl, allyl) and [PPh(4)][MoS(3)(S(t)Bu)]. AB - Intermediates in the condensation process of [MS(4)](2)(-) (M = Mo, W) to polythiometalates, in the presence of alkyl halides, had not been reported prior to our communication of [PPh(4)][WS(3)(SEt)] (Boorman, P. M.; Wang, M.; Parvez, M. J. Chem. Soc., Chem. Commun. 1995, 999-1000). We now report the isolation of a range of related compounds, with 1 degrees, 2 degrees, and 3 degrees alkyl thiolate ligands, including one Mo example. [PPh(4)][WS(3)(SR)] (R = (i)Bu (1), (i)Pr (2), (t)Bu (3), benzyl (5), allyl (6)) and [PPh(4)][MoS(3)(S(t)Bu)] (4) have been isolated in fair to good yields from the reaction of [PPh(4)](2)[MS(4)] with the appropriate alkyl halide in acetonitrile and subjected to analysis by X ray crystallography. Crystal data are as follows: for 1, triclinic space group P1 (No. 2), a = 11.0377(6) A, b = 11.1307(5) A, c = 13.6286(7) A, alpha = 82.941(1) degrees, beta = 84.877(1) degrees, gamma = 60.826(1) degrees, Z = 2; for 2, monoclinic space group P2(1)/c (No. 14), a = 9.499(6) A, b = 15.913(5) A, c = 18.582(6) A, beta = 99.29(4) degrees, Z = 4; for 3, monoclinic space group P2(1)/n (No. 14), a = 10.667(2) A, b = 17.578(2) A, c = 16.117(3) A, beta = 101.67(1) degrees, Z = 4; for 4, monoclinic space group P2(1)/n (No. 14), a = 10.558(3) A, b = 17.477(3) A, c = 15.954(3) A, beta = 101.18(2) degrees, Z = 4; for 5, monoclinic space group P2(1)/n (No. 14), a = 16.2111(9) A, b = 11.0080(6) A, c = 18.1339(10) A, beta = 111.722(1) degrees, Z = 4; for 6, triclinic space group P1 (No. 2), a = 9.4716(9) A, b = 10.4336(10) A, c = 14.4186(14) A, alpha = 100.183(2) degrees, beta = 90.457(2) degrees, gamma = 91.747(2) degrees, Z = 2. Structures 3 and 4 are isomorphous, and 1 exhibits disorder about the tertiary carbon. 6 has been shown to exhibit fluxionality in solution by variable temperature (1)H NMR studies, and an allyl migration mechanism is implicated in this process. The kinetics for the reaction of [WS(4)](2)(-) and EtBr were measured and suggest an associative nucleophilic substitution (S(N)2) mechanism. The decomposition of the [WS(3)(SEt)](-) ion is shown to be second order with respect to this ion, suggesting the formation of a transient binuclear intermediate. M-S bond cleavage is the predominant step in decomposition of 1-6 to yield alkyl sulfides, alkyl thiols, and polythiometalates such as [PPh(4)](2)[M(3)S(9)]. In contrast, reactions of [PPh(4)](2)[WO(x)()S(4)(-)(x)()] (x = 1, 2) with (t)BuBr result in the additional decomposition product of isobutene, presumably by C-S bond cleavage and beta-hydrogen transfer. Interestingly, the reaction of [PPh(4)](2)[WOS(3)] with BzCl yields 5 as the only isolable W thiolate species. PMID- 11399186 TI - Biladienones from the photooxidation of a meso-gem-disubstituted phlorin: crystal and molecular structures of the 3N + O coordinated nickel(II) and copper(II) complexes. AB - The photooxidation of a meso-gem-disubstituted phlorin gave two isomeric biladienones in an equilibrium involving a Z-E double bond photoisomerism. The structures of these bile pigments were elucidated using NMR techniques and show terminal benzoyl and pyrrolone moieties. Complexation with divalent cations (nickel(II), copper (II)) gave stable compounds whose crystal and molecular structure could be determined by X-ray diffraction. The metal is coordinated with three pyrrole nitrogen atoms and one oxygen atom of a terminal benzoyl group. In the crystal, the molecules are arranged in pairs through hydrogen bonds between the free terminal pyrrolones. PMID- 11399187 TI - Self-assembly organometallic squares with terpyridyl metal complexes as bridging ligands. AB - A series of novel heterometallic square complexes with the general molecular formulas [fac-Br(CO)(3)Re[mu-(pyterpy)(2)M]](4)(PF(6))(8) and [(dppf)Pd[mu (pyterpy)(2)Ru]](4)(PF(6))(8)(OTf(8) (4), where M = Fe (1), Ru (2), or Os (3), pyterpy is 4'-(4' "-pyridyl)-2,2':6',2' '-terpyridine, dppf = 1,1' bis(diphenylphosphino)ferrocene and OTf is trifluoromethanesulfonate, were prepared by self-assembly between BrRe(CO)(5) or (dppf)Pd(H(2)O)(2)(OTf)(2) and (pyterpy)(2)M(PF(6))(2). The obtained NMR spectra, IR spectra, electrospray ionization mass spectra, and elemental analyses are all consistent with the proposed square structures incorporating terpyridyl metal complexes as bridging ligands. Multiple redox processes were observed in all square complexes. All four complexes display strong visible absorptions in the region 400-600 nm, which are assigned as metal (Fe, Ru, or Os)-to-ligand (pyterpy) charge transfer (MLCT) bands. Square 3 exhibits an additional weak band at 676 nm, which is assigned to an Os-based (3)MLCT band. For each complex, the bands centered between 279 and 377 nm are assigned as pyterpy-based pi-pi bands and the Re-based MLCT band. Square 3 is luminescent in room-temperature solution, while squares 1, 2, and 4 do not have any detectable luminescence under identical experimental conditions. PMID- 11399188 TI - Gas-phase properties and fragmentation behavior of cationic, dinuclear iron chloride clusters Fe(2)Cl(n)()(+) (n = 1-6). AB - Sector-field mass spectrometry is used to probe the fragmentation patterns of cationic dinuclear iron chloride clusters Fe(2)Cl(n)()(+) (n = 1-6). For the chlorine-rich, high-valent Fe(2)Cl(n)()(+) ions (n = 4-6), losses of atomic and molecular chlorine prevail in the unimolecular and collision-induced dissociation patterns. Instead, the chlorine deficient, formally low-valent Fe(2)Cl(n)()(+) clusters (n = 1-3) preferentially undergo unimolecular degradation to mononuclear FeCl(m)()(+) ions. In addition, photoionization is used to determine IE(Fe(2)Cl(6)) = 10.85 +/- 0.05 eV along with appearance energy measurements for the production of Fe(2)Cl(5)(+) and Fe(2)Cl(4)(+) cations from iron(III) chloride vapor. The combination of the experimental results allows an evaluation of some of the thermochemical properties of the dinuclear Fe(2)Cl(n)()(+) cations: e.g., Delta(f)H(Fe(2)Cl(+)) = 232 +/- 15 kcal/mol, Delta(f)H(Fe(2)Cl(2)(+)) = 167 +/- 4 kcal/mol, Delta(f)H(Fe(2)Cl(3)(+)) = 139 +/- 4 kcal/mol, Delta(f)H(Fe(2)Cl(4)(+)) = 113 +/- 4 kcal/mol, Delta(f)H(Fe(2)Cl(5)(+)) = 79 +/- 5 kcal/mol, and Delta(f)H(Fe(2)Cl(6)(+)) = 93 +/- 2 kcal/mol. The analysis of the data suggests that structural effects are more important than the formal valency of iron as far as the Fe-Cl bond strengths in the Fe(2)Cl(n)()(+) ions are concerned. PMID- 11399189 TI - Variable coordination modes of hydrotris(3-isopropyl-4-bromopyrazolyl)borate (Tp') in Fe(II), Mn(II), Cr(II), and Cr(III) Complexes: formation of MTp'Cl (M = Fe and Mn), structural isomerism in CrTp'(2), and the observation of Tp' (-) as an uncoordinated anion. AB - The syntheses of the 4-coordinate Tp'MCl complexes (where M = Fe (1), Mn (2); and Tp' = hydrotris(3-isopropyl-4-bromopyrazolyl)borate) are described. The single crystal X-ray structures show that the metal centers have distorted tetrahedral coordination. Analogous reaction of CrCl(2)(MeCN)(2) with TlTp' gave Cr(kappa(3) Tp')(kappa(2)-Tp') (3) as the initial product. The 5-coordinate structure was assigned by single-crystal X-ray crystallography, and it was found that the kappa(3) ligand had isomerized to hydro(3-isopropyl-4-bromopyrazolyl)(2)(5 isopropyl-4-bromopyrazolyl)borate). 3 is labile in solution: in pentane it slowly converts to the 6-coordinate isomer Cr(kappa(3)-Tp')(2) (4), whose structure was determined by X-ray crystallography. In 4 both ligands are isomerized. Both 3 and 4 display Jahn-Teller distorted structures expected for high-spin d(4) configurations. Variable temperature magnetic susceptibility measurements confirm that 1, 2, and 3 all have high-spin electronic configurations in the range 5-300 K. In benzene solution 3 decomposes; one product [Cr(kappa(3)-Tp')(2)](+)[Tp'](-) (5), was identified by X-ray crystallography. 5 contains a pseudooctahedral Cr(III) cation with both ligands in the isomerized form and an uncoordinated Tp' ligand as counterion. Mechanistic studies reveal that this reaction is light rather than heat induced. IR spectroscopy is utilized to confirm the ligand hapticity in all complexes from the value of nu(B)(-)(H), and comparison is made with similar compounds. PMID- 11399190 TI - Synthesis and properties of [Ru(2)(acac)(4)(bptz)](n+) (n=0,1) and crystal structure of [Ru(2)(acac)(4)(bptz)]. AB - The neutral complex [Ru(2)(acac)(4)(bptz)] (I) has been prepared by the reaction of Ru(acac)(2)(CH(3)CN)(2) with bptz (bptz = 3,6-bis(2-pyridyl)-1,2,4,5 tetrazine) in acetone. The diruthenium(II,II) complex (I) is green and exhibits an intense metal-ligand charge-transfer band at 700 nm. Complex I is diamagnetic and has been characterized by NMR, optical spectroscopy, IR, and single-crystal X ray diffraction. Crystal structure data for I are as follows: triclinic, P1, a = 11.709(2) A, b = 13.487(3) A, c = 15.151(3) A, alpha = 65.701(14) degrees, beta = 70.610(14) degrees, gamma = 75.50(2) degrees, V = 2038.8(6) A(3), Z = 2, R = 0.0610, for 4397 reflections with F(o) > 4sigmaF(o). Complex I shows reversible Ru(2)(II,II)-Ru(2)(II,III) and Ru(2)(II,III)-Ru(2)(III,III) couples at 0.17 and 0.97 V, respectively; the 800 mV separation indicates considerable stabilization of the mixed-valence species (K(com) > 10(13)). The diruthenium(II,III) complex, [Ru(2)(acac)(4)(bptz)](PF(6)) (II) is prepared quantitatively by one-electron oxidation of I with cerium(IV) ammonium nitrate in methanol followed by precipitation with NH(4)PF(6). Complex II is blue and shows an intense MLCT band at 575 nm and a weak band at 1220 nm in CHCl(3), which is assigned as the intervalence CT band. The mixed valence complex is paramagnetic, and an isotropic EPR signal at g = 2.17 is observed at 77 and 4 K. The solvent independence and narrowness of the 1200 nm band show that complex II is a Robin and Day class III mixed-valence complex. PMID- 11399191 TI - Models for extradiol cleaving catechol dioxygenases: syntheses, structures, and reactivities of iron(II)-monoanionic catecholate complexes. AB - Crystallographic and spectroscopic studies of extradiol cleaving catechol dioxygenases indicate that the enzyme-substrate complexes have both an iron(II) center and a monoanionic catecholate. Herein we report a series of iron(II) monoanionic catecholate complexes, [(L)Fe(II)(catH)](X) (1a, L = 6-Me(3)-TPA (tris(6-methyl-2-pyridylmethyl)amine), catH = CatH (1,2-catecholate monoanion); 1b, L = 6-Me(3)-TPA, catH = DBCH (3,5-di-tert-butyl-1,2-catecholate monoanion); 1c, L = 6-Me(2)-bpmcn (N,N'-dimethyl-N,N'-bis(6-methyl-2-pyridylmethyl)-trans-1,2 diaminocyclohexane), catH = CatH; 1d, L = 6-Me(2)-bpmcn, catH = DBCH), that model such enzyme complexes. The crystal structure of [(6-Me(2)-bpmcn)Fe(II)(DBCH)](+) (1d) shows that the DBCH ligand binds to the iron asymmetrically as previously reported for 1b, with two distinct Fe-O bonds of 1.943(1) and 2.344(1) A. Complexes 1 react with O(2) or NO to afford blue-purple iron(III)-catecholate dianion complexes, [(L)Fe(III)(cat)](+) (2). Interestingly, crystallographically characterized 2d, isolated from either reaction, has the N-methyl groups in a syn configuration, in contrast to the anti configuration of the precursor complex, so epimerization of the bound ligand must occur in the course of isolating 2d. This notion is supported by the fact that the UV-vis and EPR properties of in situ generated 2d(anti) differ from those of isolated 2d(syn). While the conversion of 1 to 2 in the presence of O(2) occurs without an obvious intermediate, that in the presence of NO proceeds via a metastable S = (3)/(2) [(L)Fe(catH)(NO)](+) adduct 3, which can only be observed spectroscopically but not isolated. Intermediates 3a and 3b subsequently disproportionate to afford two distinct complexes, [(6-Me(3)-TPA)Fe(III)(cat)](+) (2a and 2b) and [(6-Me(3) TPA)Fe(NO)(2)](+) (4) in comparable yield, while 3d converts to 2d in 90% yield. Complexes 2b and anti-2d react further with O(2) over a 24 h period and afford a high yield of cleavage products. Product analysis shows that the products mainly derive from intradiol cleavage but with a small extent of extradiol cleavage (89:3% for 2b and 78:12% for anti-2d). The small amounts of the extradiol cleavage products observed may be due to the dissociation of an alpha-methyl substituted pyridyl arm, generating a complex with a tridentate ligand. Surprisingly, syn-2d does not react with O(2) over the course of 4 days. These results suggest that there are a number of factors that influence the mode and rate of cleavage of catechols coordinated to iron centers. PMID- 11399192 TI - Spiral, herringbone, and triple-decker silver(I) complexes of benzopyrene derivatives assembled through eta(2)-coordination. AB - Three novel silver(I) complexes with benzopyrene derivatives were synthesized and characterized in this paper. Treatment of AgClO(4)*H(2)O with 7 methylbenzo[a]pyrene (L(1)) afforded [Ag(2)(L(1))(toluene)(0.5)(ClO(4))(2)](n)() (1) which exhibits a 2-D sheet structure with double-stranded helical motifs. Reaction of AgCF(3)SO(3) with dibenzo[b,def ]chrysene (L(2)) gave rise to an unprecedented cocrystallization structure, ([Ag(2)(L(2))(CF(3)SO(3))(2)][Ag(2)(toluene)(2)(CF(3)SO(3))(2)])(n)() (2), formed by a 2-D neutral lamellar polymer and a 1-D neutral rodlike one. The ligand benzo[e]pyrene (L(3)) coordinated to silver(I) ions generating a closed triple decker tetranuclear complex [Ag(4)(L(3))(4)(p-xylene)(ClO(4))(4)] (3) which can be regarded as a stacking polymer owing to existing intermolecular pi-pi stack interactions. The structural diversity of the silver(I) coordination polymers with polycyclic aromatic hydrocarbons is not only related to the stacking patterns of free polycyclic aromatic hydrocarbons in the crystalline state, but also the geometric shapes of the molecules for these free ligands. In addition, the coordination of solvents to metal ions plays a crucial role in the formation of the unprecedented coordination polymeric architectures. The ESR spectroscopic results, conductivity, and synthesis properties are also discussed. PMID- 11399193 TI - Characterization of imidazolate-bridged dinuclear and mononuclear hydroperoxo complexes. AB - Dinucleating ligands having two metal-binding sites bridged by an imidazolate moiety, Hbdpi, HMe(2)bdpi, and HMe(4)bdpi (Hbdpi = 4,5-bis(di(2 pyridylmethyl)aminomethyl)imidazole, HMe(2)bdpi = 4,5-bis((6-methyl-2 pyridylmethyl)(2-pyridylmethyl)aminomethyl)imidazole, HMe(4)bdpi = 4,5-bis(di(6 methyl-2-pyridylmethyl)aminomethyl)imidazole), have been designed and synthesized as model ligands for copper-zinc superoxide dismutase (Cu,Zn-SOD). The corresponding mononucleating ligands, MeIm(Py)(2), MeIm(Me)(1), and MeIm(Me)(2) (MeIm(Py)(2) = (1-methyl-4-imidazolylmethyl)bis(2-pyridylmethyl)amine, MeIm(Me)(1) = (1-methyl-4-imidazolylmethyl)(6-methyl-2-pyridylmethyl)(2 pyridylmethyl)amine, MeIm(Me)(2) = (1-methyl-4-imidazolyl-methyl)bis(6-methyl-2 pyridylmethyl)amine), have also been synthesized for comparison. The imidazolate bridged Cu(II)-Cu(II) homodinuclear complexes represented as [Cu(2)(bdpi)(CH(3)CN)(2)](ClO(4))(3).CH(3)CN.3H(2)O (1), [Cu(2)(Me(2)bdpi)(CH(3)CN)(2)](ClO(4))(3) (2), [Cu(2)(Me(4)bdpi)(H(2)O)(2)](ClO(4))(3).4H(2)O (3), a Cu(II)-Zn(II) heterodinuclear complex of the type of [CuZn(bdpi)(CH(3)CN)(2)](ClO(4))(3).2CH(3)CN (4), Cu(II) mononuclear complexes of [Cu(MeIm(Py)(2))(CH(3)CN)](ClO(4))(2).CH(3)CN (5), [Cu(MeIm(Me)(1))(CH(3)CN)](ClO(4))(2)( )()(6), and [Cu(MeIm(Me)(2))(CH(3)CN)](ClO(4))(2)( )()(7) have been synthesized and the structures of complexes 5-7 determined by X-ray crystallography. The complexes 1 7 have a pentacoordinate structure at each metal ion with the imidazolate or 1 methylimidazole nitrogen, two pyridine nitrogens, the tertiary amine nitrogen, and a solvent (CH(3)CN or H(2)O) which can be readily replaced by a substrate. The reactions between complexes 1-7 and hydrogen peroxide (H(2)O(2)) in the presence of a base at -80 degrees C yield green solutions which exhibit intense bands at 360-380 nm, consistent with the generation of hydroperoxo Cu(II) species in all cases. The resonance Raman spectra of all hydroperoxo intermediates at -80 degrees C exhibit a strong resonance-enhanced Raman band at 834-851 cm(-1), which shifts to 788-803 cm(-1) (Deltanu = 46 cm(-1)) when (18)O-labeled H(2)O(2) was used, which are assigned to the O-O stretching frequency of a hydroperoxo ion. The resonance Raman spectra of hydroperoxo adducts of complexes 2 and 6 show two Raman bands at 848 (802) and 834 (788), 851 (805), and 835 (789) cm(-1) (in the case of H(2)(18)O(2), Deltanu = 46 cm(-1)), respectively. The ESR spectra of all hydroperoxo complexes are quite close to those of the parent Cu(II) complexes except 6. The spectrum of 6 exhibits a mixture signal of trigonal-bipyramid and square-pyramid which is consistent with the results of resonance Raman spectrum. PMID- 11399194 TI - Synthesis of homochiral tris(2-alkyl-2-aminoethyl)amine derivatives from chiral alpha-amino aldehydes and their application in the synthesis of water soluble chelators. AB - A novel synthesis of 3-fold symmetric, homochiral tris(2-alkyl-2-aminoethyl)amine (TREN) derivatives is presented. The synthesis is general in scope, starting from readily prepared chiral alpha-amino aldehydes. The optical purity of the N-BOC protected derivatives of tris(2-methyl-2-aminoethyl)amine and tris(2 hydroxymethyl-2-aminoethyl)amine has been ascertained by polarimetry and chiral NMR chemical shift experiments. An X-ray diffraction study of the L-alanine derivative (tris(2-methyl-2-aminoethyl)amine.3 HCl, L-Ala(3)-TREN) is presented: crystals grown from ether diffusion into methanol are cubic, space group P2(1)3 with unit cell dimensions a = 11.4807(2) A, V = 1513.23(4) A(3), and Z = 4. Attachment of the triserine derived backbone tris(2-hydroxymethyl-2 aminoethyl)amine (L-Ser(3)-TREN) to three 3-hydroxy-1-methyl-2(1H)-pyridinonate (3,2-HOPO) moieties, followed by complexation with Gd(III) gives the complex Gd(L Ser(3)-TREN-Me-3,2-HOPO)(H(2)O)(2), which is more water soluble than the parent Gd(TREN-Me-3,2-HOPO)(H(2)O)(2) and a promising candidate for magnetic resonance imaging (MRI) applications. Crystals of the chiral ferric complex Fe(L-Ser(3) TREN-Me-3,2-HOPO) grown from ether/methanol are orthorhombic, space group P2(1)2(1)2(1), with unit cell dimensions a = 13.6290(2) A, b = 18.6117(3) A, c = 30.6789(3) A, V = 7782.0(2) A(3), and Z = 8. The solution conformation of the ferric complex has been investigated by circular dichroism spectroscopy. The coordination chemistry of this new ligand and its iron(III) and gadolinium(III) complexes has been studied by potentiometric and spectrophotometric methods. Compared to the protonation constants of previously studied polydentate 3,2-HOPO 4-carboxamide ligands, the sum of protonation constants (log beta(014)) of L Ser(3)-TREN-Me-3,2-HOPO (24.78) is more acidic by 1.13 log units than the parent TREN-Me-3,2-HOPO. The formation constants for the iron(III) and gadolinium(III) complexes have been evaluated by spectrophotometric pH titration to be (log K) 26.3(1) and 17.2(2), respectively. PMID- 11399195 TI - Coordination chemistry of Rh(III) porphyrins: complexes with hydrazine, disulfide, and diselenide bridging ligands. AB - Rh(III) porphyrin complexes with bridging hydrazine and substituted hydrazine ligands were characterized in solution by (1)H NMR spectroscopy and in the solid state by X-ray diffraction. Addition of further ligand to these species afforded 1:1 complexes in which methylhydrazine and N,N-dimethylhydrazine preferentially bound to the Rh center through the substituted nitrogen atom, as evidenced by (1)H NMR chemical shifts. An alkylated Rh(III) porphyrin was isolated as a decomposition product of the reaction of N,N-dimethylhydrazine with Rh(III) porphyrin in the presence of light and oxygen. Me(2)Se(2) and Me(2)S(2) formed bridging and nonbridging complexes with Rh(III) porphyrin, analogous to that observed with N,N'-dimethylhydrazine. PMID- 11399196 TI - A general high-yield route to bis(salicylaldimine) zinc(II) complexes: application to the synthesis of pyridine-modified salen-type zinc(II) complexes. AB - A general, direct, and high-yield synthesis of bis(salicylaldimine) zinc complexes from the ligands and Et(2)Zn is reported. This synthetic method is particularly valuable, not only because it allows the efficient preparation of salen-type complexes of zinc but also because it can be used to prepare bifunctional pyridine-modified zinc(II) bis(salicylidene) complexes, which are potentially useful compounds for applications in asymmetric catalysis and materials chemistry. The synthesis and complete structural characterization of a new series of pyridine-modified zinc(II) bis(salicylidene) ligands is discussed. PMID- 11399197 TI - Equilibrium and kinetics studies of reactions of manganese acetate, cobalt acetate, and bromide salts in acetic acid solutions. AB - The oxidation of hydrogen bromide and alkali metal bromide salts to bromine in acetic acid by cobalt(III) acetate has been studied. The oxidation is inhibited by Mn(OAc)(2) and Co(OAc)(2), which lower the bromide concentration through complexation. Stability constants for Co(II)Br(n)() were redetermined in acetic acid containing 0.1% water as a function of temperature. This amount of water lowers the stability constant values as compared to glacial acetic acid. Mn(II)Br(n)() complexes were identified by UV-visible spectroscopy, and the stability constants for Mn(II)Br(n)() were determined by electrochemical methods. The kinetics of HBr oxidation shows that there is a new pathway in the presence of M(II)Br(n)(). Analysis of the concentration dependences shows that CoBr(2) and MnBr(2) are the principal and perhaps sole forms of the divalent metals that react with Co(III) and Mn(III). The interpretation of these data is in terms of this step (M, N = Mn or Co): M(OAc)(3) + N(II)Br(2) + HOAc --> M(OAc)(2) + N(III)Br(2)OAc. The second-order rate constants (L mol(-)(1) s(-)(1)) for different M, N pairs in glacial acetic acid are 4.8 (Co, Co at 40 degrees C), 0.96 (Mn, Co at 20 degrees C), 0.15 (Mn(III).Co(II), Co at 20 degrees C), and 0.07 (Mn, Mn at 20 degrees C). Following that, reductive elimination of the dibromide radical is proposed to occur: N(III)Br(2)OAc + HOAc --> N(OAc)(2) + HBr(2)(*). This finding implicates the dibromide radical as a key intermediate in this chemistry, and indeed in the cobalt-bromide catalyzed autoxidation of methylarenes, for which some form of zerovalent bromine has been identified. The selectivity for CoBr(2) and MnBr(2) is consistent with a pathway that forms this radical rather than bromine atoms which are at a considerably higher Gibbs energy. Mn(OAc)(3) oxidizes PhCH(2)Br, k = 1.3 L mol(-)(1) s(-)(1) at 50.0 degrees C in HOAc. PMID- 11399198 TI - Design and synthesis of a sterically hindered pyridine and its encapsulation of silver(I) cation. PMID- 11399199 TI - Hydrothermal synthesis and structural characterization of a coordination polymer containing heptanuclear Co(7)(mu(3)-OH)(8) clusters. PMID- 11399200 TI - A novel LIESST iron(II) complex exhibiting a high relaxation temperature. PMID- 11399201 TI - Multinuclear NMR spectra, (1)H-T(1) relaxation, conformational behavior, and intramolecular H(delta-)....(delta+)H contacts of N-borane cyclic adducts in solution. PMID- 11399202 TI - Bowl-shaped and highly inert bis(mu-hydroxo)-platinum(II) dimer. PMID- 11399203 TI - Luminescence of dimethylgallium(III) azide. PMID- 11399204 TI - Disruption of spinal cord white matter and sciatic nerve geometry inhibits axonal growth in vitro in the absence of glial scarring. AB - BACKGROUND: Axons within the mature mammalian central nervous system fail to regenerate following injury, usually resulting in long-lasting motor and sensory deficits. Studies involving transplantation of adult neurons into white matter implicate glial scar-associated factors in regeneration failure. However, these studies cannot distinguish between the effects of these factors and disruption of the spatial organization of cells and molecular factors (disrupted geometry). Since white matter can support or inhibit neurite growth depending on the geometry of the fiber tract, the present study sought to determine whether disrupted geometry is sufficient to inhibit neurite growth. RESULTS: Embryonic chick sympathetic neurons were cultured on unfixed longitudinal cryostat sections of mature rat spinal cord or sciatic nerve that had been crushed with forceps ex vivo then immediately frozen to prevent glial scarring. Neurite growth on uncrushed portions of spinal cord white matter or sciatic nerve was extensive and highly parallel with the longitudinal axis of the fiber tract but did not extend onto crushed portions. Moreover, neurite growth from neurons attached directly to crushed white matter or nerve tissue was shorter and less parallel compared with neurite growth on uncrushed tissue. In contrast, neurite growth appeared to be unaffected by crushed spinal cord gray matter. CONCLUSIONS: These observations suggest that glial scar-associated factors are not necessary to block axonal growth at sites of injury. Disruption of fiber tract geometry, perhaps involving myelin-associated neurite-growth inhibitors, may be sufficient to pose a barrier to regenerating axons in spinal cord white matter and peripheral nerves. PMID- 11399205 TI - Myelin contributes to the parallel orientation of axonal growth on white matter in vitro. AB - BACKGROUND: Brain and spinal cord white matter can support extensive axonal growth. This growth is generally constrained to an orientation that is parallel to the longitudinal axis of the fiber tract. This constraint is presumably due to permissive and non-permissive substrates that are interleaved with each other and oriented in parallel within the tract. RESULTS: Embryonic chick sympathetic neurons were cultured on cryostat sections of rat brain and the orientation of neurite growth on white matter was assessed. To determine if haptotaxis is sufficient to guide parallel neurite growth, neurons were cultured under conditions designed to interfere with interactions between growing neurites and factors that act as biochemical contact guidance cues but not interactions with haptotactic cues. Under these conditions, neurites extending on white matter were not exclusively oriented in parallel to the fiber tract, suggesting that biochemical cues are involved. To assess the role of myelin in guiding parallel neurite growth, neurons were cultured on myelin-deficient corpus callosum. These neurons also extended neurites that were not constrained to a parallel orientation. Moreover, preincubation with NGF and treatment with cAMP analogs, manipulations that attenuate overall myelin-mediated inhibition of neurite growth, also led to a reduced parallel orientation of neurite growth. CONCLUSIONS: The present studies suggest that some of the relevant factors that constrain axonal growth on white matter are not haptotactic in nature and appear to be partly mediated by factors that are associated with myelin and may involve myelin-associated "inhibitors". PMID- 11399206 TI - The evaluation of yeast derivatives as adjuvants for the immune response to the Bm86 antigen in cattle. AB - BACKGROUND: The Gavac vaccine against the cattle tick Boophilus microplus has proven its efficacy in a large number of controlled and field experiments. However, this vaccine could be further improved by searching for new alternative adjuvants that would induce a stronger long-lasting immune response. We conducted several experiments to assay the adjuvant effect of fractions of the recombinant yeast Pichia pastoris in mouse and cattle models. In previous experiments, the combination of the yeast membrane with saponin was the most effective formulation in stimulating the humoral immune response in mice, eliciting a response higher than Montanide 888. The response was predominantly of the IgG1 isotype. Here, we evaluated the response in cattle and compared the results with that obtained in mice. RESULTS: Bm86 on the membrane of P. pastoris plus saponin produced high antibody titers in cattle, though the protection level against tick infestations was lower when compared to Gavac, probably due to a decrease in the IgG1/IgG2 ratio. The predictive value of the mouse model was studied through correlation analysis between the isotype levels in mice and the efficacy of formulations in cattle. Good correlation was established between the level of antibodies in mice and cattle, and between the amount of anti -Bm86 IgG1 in mice and the degree of protection in cattle. CONCLUSION: Mouse model have the potential to predict immunogenicity and efficacy of formulations in cattle. These results also support the use of the yeast expression system for recombinant vaccine formulations, enabling the prediction of more cost--effective formulations. PMID- 11399207 TI - Contribution of different HFE genotypes to iron overload disease: a pooled analysis. AB - PURPOSE: To determine the contribution of the C282Y and H63D mutations in the HFE gene to clinical expression of hereditary hemochromatosis. METHODS: Pooled analysis of 14 case-control studies reporting HFE genotype data, to evaluate the association of different HFE genotypes with iron overload. In addition, we used data from the pooled analysis and published data to estimate the penetrance of the C282Y/C282Y genotype. RESULTS: Homozygosity for the C282Y mutation carried the largest risk for iron overload (OR = 4383, 95% CI 1374 to >10,000) and accounted for the majority of hemochromatosis cases (attributable fraction (AF) = 0.73). Risks for other genotypes were much smaller: OR = 32 for genotype C282Y/H63D (95% CI 18.5 to 55.4, AF = 0.06); OR = 5.7 for H63D/H63D (95% CI 3.2 to 10.1, AF = 0.01); OR = 4.1 for C282Y heterozygosity (95% CI 2.9 to 5.8, with heterogeneity in study results, making this association uncertain); and OR = 1.6 for H63D heterozygosity (95% CI 1 to 2.6, AF = 0.03). Estimates of penetrance for the C282Y/C282Y genotype were highly sensitive to estimates of the prevalence of iron overload disease. At a prevalence of 2.5 per 1000 or less, penetrance of the C282Y/C282Y genotype is unlikely to exceed 50%. Penetrance of other HFE genotypes is much lower. CONCLUSIONS: C282Y homozygosity confers the highest risk for iron overload but the H63D mutation is also associated with increased risk. Our data indicate a gradient of risk associated with different HFE genotypes and thus suggest the presence of other modifiers, either genetic or environmental, that contribute to the clinical expression of hemochromatosis. PMID- 11399208 TI - A comparison of the Berlin and Ghent nosologies and the influence of dural ectasia in the diagnosis of Marfan syndrome. AB - PURPOSE: To compare the Berlin and Ghent diagnostic criteria for Marfan syndrome and evaluate the utility of screening for dural ectasia in the diagnosis of Marfan syndrome. METHODS: Review of clinical and radiographic data on 73 patients evaluated for Marfan syndrome at the National Institutes of Health. RESULTS: Nineteen percent of patients diagnosed under the Berlin criteria failed to meet the Ghent standard. Dural ectasia was the second most common major diagnostic manifestation, and screening for dural ectasia established the diagnosis of Marfan syndrome in 23% of patients under the Ghent criteria. CONCLUSIONS: Some patients are appropriately excluded from the diagnosis of Marfan syndrome by the Ghent criteria. Determination of dural ectasia is valuable in the diagnosis of Marfan syndrome. PMID- 11399209 TI - Clinical heterogeneity in autosomal dominant optic atrophy in two 3q28-qter linked central Illinois families. AB - PURPOSE: To examine the clinical and genetic heterogeneity of autosomal dominant optic atrophy among two unrelated central Illinois families. METHODS: Forty-three individuals from two pedigrees had complete eye examinations. Linkage analysis was performed with microsatellite markers from the region 3q28-29. RESULTS: Visual acuity in 21 affected individuals ranged from 20/25 to 20/800. Vision loss was more severe in males than females (P = 0.02). Color vision testing revealed generalized dyschromatopsia. Both visual acuity and color vision deteriorated with age. Linkage was established to chromosome 3q28-29 (LODmax = 4.68 for D3S2305). CONCLUSION: Autosomal dominant optic atrophy linked to chromosome 3q28 29 shows intrafamilial phenotypic variation as well as sex-influenced severity in two Midwestern families. PMID- 11399210 TI - Acute intermittent porphyria: novel missense mutations in the human hydroxymethylbilane synthase gene. AB - PURPOSE: To identify mutations in families with acute intermittent porphyria, an autosomal dominant inborn error of metabolism that results from the half-normal activity of the third enzyme in the heme biosynthetic pathway, hydroxymethylbilane synthase. METHODS: Mutations were identified by direct solid phase sequencing. RESULTS: Two novel missense mutations E80G and T78P and three previously reported mutations, R173W, G111R, and the splice site lesion, IVS1+1, were detected, each in an unrelated proband. The causality of the novel missense mutations was demonstrated by expression studies. CONCLUSION: These findings provide for the precise diagnosis of carriers in these families and further expand the molecular heterogeneity of AIP. PMID- 11399211 TI - Alcohol problems: Is there a specifically French view? PMID- 11399212 TI - Recent developments in drug-testing arrestees for research purposes: an English perspective on an international phenomenon. PMID- 11399213 TI - Attention deficit hyperactivity disorder and substance use disorders: Is there a causal link? AB - Attention-deficit hyperactivity disorder (ADHD), characterized by restless, inattentive and hyperactive behaviours, is a relatively common childhood disorder that affects approximately 5% of the general population. There has been controversy about whether ADHD increases risks of developing substance use disorders. The available evidence suggests that, in the absence of conduct disorder, ADHD is not associated with an increased risk of substance use problems in males. There is only limited evidence on the role of ADHD in the aetiology of substance use disorders among females. While ADHD has traditionally been considered as a childhood disorder, it may also occur in adults; research needs to examine the extent to which ADHD in adulthood increases the risk of substance use disorders. PMID- 11399214 TI - Examining the limits of the buprenorphine interdosing interval: daily, every third-day and every-fifth-day dosing regimens. AB - AIMS: Opioid-dependent outpatients may be more likely to present for pharmacological treatment if less than daily dosing can be arranged. These studies compared opioid withdrawal symptoms during 24-, 72-, and 120-hour buprenorphine dosing regimens and evaluated participants' preferences for these different dosing regimens. PARTICIPANTS: Thirty-three opioid-dependent participants received daily sublingual maintenance doses of 4 mg/70 kg (n = 14) or 8 mg/70 kg (n = 19) of liquid buprenorphine. METHODS: In Study I participants received, in a random order, three dosing regimens for five repetitions of each: daily maintenance doses every 24 hours (4 or 8 mg/70 kg), triple the daily maintenance dose every 72 hours (12 or 24 mg/70 kg) and quintuple the daily maintenance dose every 120 hours (20 or 40 mg/70 kg). Doses were administered under double-blind procedures, and placebos were administered on the interposed days during the latter two regimens. Subjective and observer ratings of opioid withdrawal symptoms were assessed daily prior to receipt of each dose. In Study II, a new group of participants received each of the three dosing regimens under open-dosing procedures and then chose between the different dosing regimens. FINDINGS: Opioid withdrawal symptoms increased significantly during the every fifth-day dosing regimen in both the blind- and open-dosing studies. In the choice phase of Study II, only one participant (7%) chose quintuple-every-fifth day dosing over all other dosing options. CONCLUSIONS: These results suggest that the maximum duration of action of buprenorphine is less than 5 days when five times the daily maintenance dose is provided. PMID- 11399215 TI - Comparison of individual and group cognitive-behavioral therapy for alcohol and/or drug-dependent patients. AB - AIMS AND DESIGN: A randomized clinical trial was performed to evaluate the influence of two formats of cognitive-behavioral psychotherapy (individual vs. group) in the treatment of alcohol and/or drug dependent patients. SETTING: Public outpatient drug dependence service. PARTICIPANTS: One hundred and fifty five alcohol and/or drug-dependent patients. INTERVENTION: The patients were randomly assigned to individual (n = 77) or group (n = 78) treatment formats. The treatment was developed into two phases: acquisition (eight sessions) and maintenance (nine sessions), distributed over an 8-month period. MEASUREMENTS: Alcohol and drug use, severity of dependence, and alcohol- and drug-related problems were evaluated at pre-treatment and 15 months after admission to treatment. FINDINGS: At follow-up evaluation both groups of patients presented similar levels of drug consumption, dependence and associated problems. Although group-treated patients reported slightly higher levels of alcohol consumption (both at baseline and follow-up) differences between the formats disappear if baseline levels are included as covariates. Compliance with treatment and a measure of drug severity were predictors of success for the drug dependents. The number of sessions attended and high GGT levels at admission were positively correlated with success for the alcohol dependents. CONCLUSIONS: The two modalities presented similar outcomes and, as the group format could present a better cost-benefit ratio, it may be used without decreasing compliance with treatment or treatment effectiveness. PMID- 11399216 TI - Potential risk factors for the transition to injecting among non-injecting heroin users: a comparison of former injectors and never injectors. AB - AIMS: To compare potential risk factors for the transition to injecting among non injecting heroin users (NIUs) with different injecting histories. DESIGN: Cross sectional data from baseline structured interviews with NIUs in a study on transitions to injecting. Sample recruited by outreach or chain-referral in New York City (NYC), 1996-1998. SETTING: Recruitment of sample and interviews conducted in a NYC neighborhood where many drug users reside and/or use drugs. PARTICIPANTS: Of 575 NIUs, 67% had never injected; 16% had injected one to nine times (infrequent former injectors (IFI)); and 18% 10 or more times (frequent former injectors (FFI)). MEASUREMENTS: Controlling for age and race/ethnicity, adjusted odds ratios were estimated in multivariate logistic regression, and differences in means tested by ANCOVA. FINDINGS: FFI (compared to never injectors and IFI) were more likely: to be homeless; to be unemployed; to be long-time users; to be younger at first heroin use; to not have initiated heroin use through non-injected routes; to not be afraid of injecting themselves with needles; to sniff heroin with former IDUs; and, for both men and women separately, to have sex partners who were former IDUs. Both FFI and IFI were twice as likely as never injectors to perceive that their friends thought that it was "OK" to inject drugs. CONCLUSIONS: FFI have multiple individual and network characteristics that may increase their risk of injecting drugs. Interventions among NIUs to prevent transitions to injecting need to ascertain NIUs' injecting history and address the many potential risks that FFI have for resuming injecting drug use. PMID- 11399217 TI - Psychopathology in adolescence predicts substance use in young adulthood. AB - AIMS: To investigate prospective associations between psychopathology in adolescence and tobacco, alcohol and drug use in young adulthood. DESIGN: A sample of 787 10-14-year-olds from the Dutch general population was prospectively followed-up across an 8-year interval. The Child Behavior Checklist (CBCL) was administered at initial assessment, and at 2- and 4-year follow-ups. Substance abuse was assessed with the Young Adult Self-Report (YASR) at 8-year follow-up. FINDINGS: The Thought Problems scale of the CBCL was the strongest predictor of alcohol use, while smoking was predicted by the Thought Problems and Delinquent Behavior scales. The strongest association with drug use in young adulthood was for the CBCL Delinquent Behavior scale. Predictive value of predictors in early adolescence was as important as in late adolescence. CONCLUSION: To investigate pathways towards substance use in young adulthood, studies assessing a broad range of possible predictors, including Thought Problems, at different developmental stages of adolescence, are needed. PMID- 11399218 TI - Gender differences, physiological arousal and the role of winning in fruit machine gamblers. AB - AIMS: To examine gender differences in changes in physiological arousal as indexed by heart rate during fruit machine gambling while controlling for the confounding effect of movement and as a function of winning and losing, and to examine relationships between sensation-seeking, self-reported arousal during gambling, heart rate during gambling and loss of control of gambling behaviour. SAMPLE, DESIGN AND MEASUREMENTS: Heart rate and subjective arousal were recorded in a sample of 22 male and 20 female fruit machine players before, during and after the gambling process. At baseline measures were taken of sensation seeking, self-reported arousal during gambling and loss of control of gambling. A simulation of the behaviour used to operate fruit machines was used during baseline measurement. FINDINGS: Significant increases in heart-rate over movement controlled baselines were observed within participants during gambling. However, for the losing group these differences were slight compared to the increases found when participants won during play. Furthermore, winning/losing was the only variable among a range of variables which was a significant predictor of heart rate increase during play over a baseline control. Additionally HR levels of males and females behaved the same way in relation to the manipulations. Significant correlations between HR during play and the measures of both subjective arousal and sensation seeking were also present. CONCLUSIONS: Winning during gambling is essential in order to maintain increased HR levels over baseline after play. Subjective arousal and sensation seeking may be predictors of levels of arousal experienced during play. PMID- 11399219 TI - The effectiveness of callback counselling for smoking cessation: a randomized trial. AB - AIMS: The development of acceptable, widely available and effective smoking cessation methods is central to public health strategy for tobacco control. We examined the effectiveness of a telephone callback counselling intervention, compared to the provision of self-help resources alone. METHODS: Participants were 998 smokers calling a state-wide "Quitline" service randomly allocated to either callback counselling or ordinary care. The callback condition consisted of a series of brief counselling calls at strategic times in addition to ordinary care. The number of calls varied according to caller needs, and most occurred generally just before the person's quit day and in the week or two after it. The service was delivered by trained telephone counsellors. RESULTS: At the 3-month follow-up, significantly more participants in the callback group (24%) reported that they were quit, compared to those in the usual care comparison group (13%). The difference in point prevalence of smoking declined to 6% by the 12-month follow-up. Using sustained abstinence there was a significant benefit of callback counselling at 12-month follow-up. Treating dropouts as smokers reduced the overall magnitude of the effects somewhat. The benefit of callbacks was to marginally increase quit attempts and to significantly reduce relapse. CONCLUSION: Our findings are consistent with those of other studies demonstrating benefits of callback telephone counselling to facilitate cessation. Such counselling provides a flexible, relatively inexpensive and widely available form of cessation service. It appears to encourage a greater proportion of quit attempts and to reduce the rate of relapse among those quitting. Further research is required to determine ways to enhance effectiveness, particularly studies of how to reduce relapse. PMID- 11399220 TI - Smoking cessation and smoking patterns in the general population: a 1-year follow up. AB - OBJECTIVE: To assess the prevalence of motivation and behaviours relating to smoking cessation and attempts at harm minimization and the stability of these over a 1-year period; to identify demographic, social, behavioural and psychological predictors of attempts to stop smoking and the success of these attempts. DESIGN: Face-to-face interviews were carried out with a national sample of UK smokers in April/May 1996 with follow-up 1 year later. SUBJECTS: The original response rate was 61% (1478 of 1911 adult smokers), and of these 1012 were followed-up 1 year later (68% of those who were originally contactable). RESULTS: Thirty-one per cent of smokers reported making at least one quit attempt during the follow-up period and 17% made a quit attempt in the first 9 months of that period. Of these 29% were still not smoking at least 3 months later. Fifty one per cent of smokers had tried to cut down in the year leading up to the first survey. There was a fair degree of consistency over time in individual smokers' desires and intentions to stop smoking across both surveys and in the incidence of quit attempts and attempts to cut down. Beliefs about the effects of smoking on future health and having a partner who disliked their smoking were positively associated with making a quit attempt at follow-up while reporting enjoying smoking at baseline was negatively associated with making a quit attempt at follow-up. Time to first cigarette of the day and age of starting smoking were positively associated with success of quit attempts. CONCLUSIONS: Motivation and behaviours relating to smoking cessation are prevalent and fairly stable over time. Different factors appear to be related to attempts to stop and the success of those attempts. Interventions to increase smoking cessation in the population should take account of this. PMID- 11399221 TI - The clinical course of alcohol dependence associated with a low level of response to alcohol. AB - AIMS: To evaluate the clinical course of specific alcohol-related life problems and the risk for dependence on illicit drugs in individuals with relatively low and high levels of response (LR) to alcohol earlier in life. SUBJECTS: From among 439 men who were part of the 15-year follow-up of sons of alcoholics and controls, 108 were identified as having fulfilled criteria for DSM-III-R alcohol dependence. MEASURES: The LR to alcohol was originally evaluated following the consumption of 0.61 g/kg of ethanol at age 20 by determining the levels of change in subjective feelings of intoxication, body sway and several hormones such as cortisol. From the 453 original subjects, 450 completed a face-to-face 10-year follow-up evaluation, and 439 completed the 15-year protocol. FINDINGS: A comparison of the clinical course of 50 alcohol-dependent men with clearly low LR values at age 20 with that for 42 individuals whose LR scores were above the median revealed few differences. Those with a low LR had a slightly earlier age of onset of alcohol dependence (24.8 +/- 3.41 vs. 26.6 +/- 4.48 years), and this finding was unrelated to the presence of an alcohol-dependent father. Otherwise the members of the two groups demonstrated a similar course of alcohol dependence. There was no relationship between a low LR at age 20 and either the pattern of substances used or the rate of dependence on illicit drugs. CONCLUSIONS: The results indicate that for this sample a low LR to alcohol, while associated with a high risk for alcohol dependence, was not related to most aspects of the course of alcohol problems once dependence developed. PMID- 11399222 TI - Needle fixing is not a myth: comments on Al-Adwani's "The myth of needle fixation". PMID- 11399224 TI - Approaches to gene therapy for human immunodeficiency virus infection. AB - Much progress has been made in developing new and more efficient treatments for human immunodeficiency virus (HIV) infection, the cause of acquired immunodeficiency syndrome (AIDS). However, the scope of the HIV epidemic and the limitations of existing treatments necessitate the continued development of novel treatment strategies. Gene therapy is one such forward-looking strategy. Gene therapy approaches for HIV infection include efforts to interfere with viral replication directly by engineering HIV-resistant cells or indirectly by eliminating infected cells from the body, primarily by eliciting a therapeutic immune response to destroy HIV-infected cells. Although the prospect of gene therapy as a routine treatment for HIV infection remains distant, continuous progress is being made, which should also have implications for gene therapy strategies for a variety of other diseases. This article reviews some of the strategies for investigating the feasibility of gene transfer for the treatment of HIV infection. PMID- 11399225 TI - Bone marrow stem cell gene therapy of arylsulfatase A-deficient mice, using an arylsulfatase A mutant that is hypersecreted from retrovirally transduced donor type cells. AB - Arylsulfatase A (ASA)-deficient mice represent an animal model for the fatal lysosomal storage disease metachromatic leukodystrophy, which is characterized by widespread intralysosomal deposition of sulfatide. Bone marrow stem cell gene therapy in mice, using a retroviral vector mediating expression of wild-type human ASA, has the potential to ameliorate the visceral pathology, but improves the prevailing brain disease and neurologic symptoms only marginally. One factor that influences the efficacy of bone marrow transplantation therapy in lysosomal storage diseases is the secretion level of the therapeutic enzyme from donor-type cells. Here we test the potential of a hypersecreted glycosylation variant of ASA. Although this mutant lacks mannose 6-phosphate residues it is taken up by cells by a mannose 6-phosphate receptor-independent pathway and causes partial metabolic correction of ASA-deficient mouse cells. Retrovirally mediated transfer of the mutant cDNA into ASA-deficient mice results in the sustained expression of the transgene. Serum levels argue for an increased secretion of the glycosylation mutant also in vivo. Tissue levels were reduced to 2% in liver and up to 40% in kidney compared with animals treated with the wild-type enzyme, indicating reduced endocytosis. Thus, the limited uptake of the variant enzyme outweighs the putative advantageous effect of improved supply. Although the mutant enzyme is able to correct the metabolic defect partially, histological examinations did not reveal any reduction of sulfatide storage in treated animals. Surprisingly, analysis of neurologic symptoms indicated a significant improvement of the gait pattern. PMID- 11399226 TI - Differences in the human and mouse amino-terminal leader peptides of ornithine transcarbamylase affect mitochondrial import and efficacy of adenoviral vectors. AB - Mouse models of ornithine transcarbamylase (OTC) deficiency are being used to test the efficacy of viral vectors as possible vehicles for gene therapy. However, it has been demonstrated that virus containing the human OTC cDNA failed to express functional OTC enzyme in the recipient animals. Because functional OTC is assembled as a homotrimer in the mitochondria, there are at least two possible explanations for these results. Either endogenous mutant protein coassembles with the human OTC and has a "dominant-negative effect," or the human version of the protein is not appropriately imported or processed in the mouse mitochondria. To test the importance of processing, which in rodents is thought to depend on the leader peptide, adenoviral vectors containing chimeric OTC cDNAs were prepared. These vectors were evaluated in the OTC-deficient sparse fur mouse models. Although comparable levels of transgene expression were observed in all groups of mice, the only mice that had high levels of OTC activity and mitochondrial OTC immunoreactivity were those mice injected with the vectors containing the mouse leader peptide (mouse OTC and a mouse-human chimera of OTC). To address possible dominant-negative effects, adenoviruses containing mutant human or mouse OTC cDNAs were prepared and evaluated in cell lines or normal C3H mice, respectively. No inhibition of normal OTC activity was observed in either model system. Together, these studies provide no evidence of a dominant-negative effect and suggest that the human and rodent enzymes responsible for transporting of OTC and possibly other mitochondrial proteins have different specificity. PMID- 11399227 TI - A novel recombinant adeno-associated virus vaccine induces a long-term humoral immune response to human immunodeficiency virus. AB - Recombinant adeno-associated virus (AAV) has attracted tremendous interest as a promising vector for gene delivery. In this study we have developed an HIV-1 vaccine, using an AAV vector expressing HIV-1 env, tat, and rev genes (AAV-HIV vector). A single injection of the AAV-HIV vector induced strong production of HIV-1-specific serum IgG and fecal secretory IgA antibodies as well as MHC class I-restricted CTL activity in BALB/c mice. The titer of HIV-1-specific serum IgG remained stable for 10 months. When AAV-HIV vector was coadministered with AAV IL2 vector, the HIV-specific cell-mediated immunity (CMI) was significantly enhanced. Boosting with AAV-HIV vector strongly enhanced the humoral response. Furthermore, the mouse antisera neutralized an HIV-1 homologous strain, and BALB/c mice immunized via the intranasal route with an AAV vector expressing the influenza virus hemagglutinin (HA) gene showed protective immunity against homologous influenza virus challenge. These results demonstrate that AAV-HIV vector immunization may provide a novel and promising HIV vaccination strategy. PMID- 11399228 TI - Successful gene therapy via intraarticular injection of adenovirus vector containing CTLA4IgG in a murine model of type II collagen-induced arthritis. AB - We previously constructed an adenovirus vector carrying a gene encoding a soluble form of fusion protein, consisting of the extracellular portion of cytotoxic lymphocyte antigen 4 (CTLA4) and the Fc portion of human immunoglobulin G1 (Adex1CACTLA4IgG). Murine type II collagen-induced arthritis (CIA) was treated with Adex1CACTLA4IgG. A single intraarticular injection of 1 x 10(5) PFU was able to support serum CTLA4IgG at more than 10 microg/ml for at least 12 weeks and was able to inhibit the CIA clinically and histologically. In contrast, intravenous, intramuscular, or subcutaneous injection of 1 x 10(5) PFU was unable to support a significant level of serum CTLA4IgG and thus was unable to inhibit the development of arthritis. Thus, we demonstrated that (1) a low-dose intraarticular injection of Adex1CACTLA4IgG was effective in delaying the onset of CIA and reducing the severity of arthritis; (2) an intraarticular (knee joint) injection of Adex1CACTLA4IgG effectively blocked the development of arthritis in distal paws; (3) the inhibitory effect of Adex1CACTLA4IgG lasted at least up to 20 weeks; (4) although serum CTLA4IgG at more than 10 microg/ml persisted for at least 12 weeks, mice treated by intraarticular injection of Adex1CACTLA4IgG were not anergic to adenovirus and were able to mount antibody responses against various antigens. PMID- 11399229 TI - Long-term engraftment of nonobese diabetic/severe combined immunodeficient mice with human CD34+ cells transduced by a self-inactivating human immunodeficiency virus type 1 vector. AB - Human hematopoietic cells with in vivo repopulating potential hold much promise as a target for corrective gene transfer for numerous inherited or acquired hematopoietic disorders. Here we demonstrate long-term hematopoietic reconstitution of nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice with human CD34(+) cells transduced by an HIV-1-based self-inactivating (SIN) vector encoding the enhanced green fluorescent protein (EGFP). Human umbilical cord CD34(+) cells were transduced (up to 76%) at a low multiplicity of infection (MOI of 5) in the absence of cytokine prestimulation. Introduction of transduced hCD34(+) cells into irradiated recipients resulted in multilineage engraftment and stable transgene expression for 18 weeks posttransplantation. Bone marrow from transplanted mice contained up to 50% hCD45(+) cells and up to 63% hCD45(+)/EGFP(+) cells. Analysis of extramedullar splenic reconstitution showed up to 13% hCD45(+) cells and up to 41% hCD45(+)/EGFP(+) cells. Analysis of human progenitor cells isolated from bone marrow of recipient animals showed equivalent percentages of EGFP(+) colony-forming cells (CFCs) by fluorescence microscopy and by PCR analysis of provirus sequences, indicating minimal transgene silencing in vivo. These findings demonstrate the utility of lentivirus based SIN vectors for hematopoietic stem cell gene transfer and provide strong support for their future clinical evaluation. PMID- 11399230 TI - Modulation of cellular responses by plasmid CD40L: CD40L plasmid vectors enhance antigen-specific helper T cell type 1 CD4+ T cell-mediated protective immunity against herpes simplex virus type 2 in vivo. AB - Engineering gene therapy vectors to modulate the immune response is an important goal. In this regard, costimulation of T cells is a critical determinant in immune activation. The costimulatory molecule CD40, expressed on antigen presenting cells, is thought to interact with CD40 ligand (CD40L) expressed on activated CD4(+) or CD8(+) T cells to further drive interleukin-2 receptor (IL 2R) expression and antigen-specific T cell expansion necessary for both class II and class I responses. To compare the specific roles of these two costimulatory molecules in immune induction in a herpes simplex virus (HSV) model, we constructed plasmid DNAs expressing CD40 and CD40L, coimmunized these molecules with a gD plasmid vaccine, and then analyzed immune modulatory effects as well as protection against lethal HSV-2 challenge. We observed that gD-specific IgG production was unaffected by these molecules. However, a higher production of IgG2a isotype was induced by CD40L coinjection, suggesting that CD40L drives immune responses towards a helper T cell type 1 (Th1) phenotype. CD40L also enhanced Th cell proliferative responses and production of Th1-type cytokines (IL 2 and IFN-gamma) and beta-chemokines (RANTES and MIP-1alpha) from splenocytes. In contrast, CD40 showed slightly increasing effects on T cell proliferation responses and cytokine and chemokine production. When animals were challenged with a lethal dose of HSV-2, CD40L-coimmunized animals exhibited a significantly enhanced survival rate, as compared with CD40 coinjection or gD DNA vaccine alone. This enhanced protection appears to be mediated by Th1-type CD4(+) T cells, as determined by in vitro and in vivo T cell subset deletion. CD40L also promoted migration of CD4(+) T cells into the muscle sites. These studies demonstrate that CD40L can play an important role in protective antigen-specific immunity in a gene-based model system through increased expansion of the CD4(+) Th1 T cell subset in vivo. PMID- 11399231 TI - Lentivirus vectors encoding both central polypurine tract and posttranscriptional regulatory element provide enhanced transduction and transgene expression. AB - Incorporation of a central polypurine tract (cPPT) and a posttranscriptional regulatory element (PRE) into lentivirus vectors provides increased transduction efficiency and transgene expression. We compared the effects of these elements individually and together on transduction efficiency and gene expression, using lentivirus vectors pseudotyped with vesicular stomatitis virus G protein (VSV-G) and encoding enhanced green fluorescent protein (GFP) and rat erythropoietin (EPO). The transduction efficiency was greater than 2-fold higher in the vector containing the PRE element, 3-fold higher in vector encoding the cPPT element, and 5-fold increased in the GFP virus containing both cPPT and PRE elements relative to the parent virus. In comparison with parent vector the mean fluorescence intensity (MFI) of GFP expression was 7-fold higher in cells transduced with virus containing PRE, 6-fold increased in cells transduced with virus containing cPPT, and 42-fold increased in GFP-virus containing both cPPT and PRE elements. EPO-virus containing a PRE element showed a nearly 5-fold increase in EPO secretion over the parent vector, and the vector encoding both PRE and cPPT showed a 65-fold increase. Thus, lentivirus vectors incorporating both PRE and cPPT showed expression levels significantly increased over the sum of the components alone, suggesting a synergistic effect. PMID- 11399232 TI - Clinical protocol. An open-label, phase I, dose-escalation study of tumor necrosis factor-alpha (TNFerade Biologic) gene transfer with radiation therapy for locally advanced, recurrent, or metastatic solid tumors. PMID- 11399233 TI - Department of Health and Human Services National Institutes of Health Recombinant DNA Advisory Committee: minutes of meeting, September 25-26, 2000. PMID- 11399237 TI - A rapid and systematic review of the clinical effectiveness and cost effectiveness of debriding agents in treating surgical wounds healing by secondary intention. AB - BACKGROUND: Most surgically sutured wounds heal without any complication. However, in some cases wound healing can be delayed due to the presence of infection or wound breakdown. This can result in the wounds becoming cavity wounds and thus necessitate healing by secondary intention. Other surgical wounds that are not sutured but left to heal by secondary intention include abscess cavities such as perianal abscesses or breast abscesses. Surgical wounds healing by secondary intention are thought to heal more slowly than wounds healing by primary intention, especially if infection is present or healing is compromised by factors such as decreased blood supply, poor nutritional status or a general suppression of the immune response. Such wounds may contain dead tissue and have a moderate or high level of exudate. Debridement involves the removal of devitalised, necrotic tissue or fibrin from a wound. There are many different methods that can be used to debride a wound, which are broadly classified as surgical/sharp, biosurgical, mechanical, chemical, enzymatic and autolytic. Although it is generally agreed that the management of surgical wounds which contain devitalised tissue and are healing by secondary intention requires debridement, it is not always clear as to what is the best method or agent to use. There is currently a large selection of products with debriding properties available on the market, which vary considerably in cost. It is important that the choice of both debriding method and product is based on the best scientific evidence available, taking into account both cost and effectiveness data. OBJECTIVES: The review had two main objectives: (1) To determine the clinical effectiveness and cost-effectiveness of debriding agents in treating surgical wounds healing by secondary intention. (2) To evaluate the clinical effectiveness and cost-effectiveness of treating patients with surgical wounds healing by secondary intention at specialised wound care clinics as compared to conventional care. The review incorporated all debriding methods and any agent that is considered to have a debriding property. METHODS The following databases were searched using strategies designed specifically for each database: MEDLINE, EMBASE, CINAHL, HMIC (Health Management Information Consortium), CCTR via the Cochrane Library, the National Research Register (NRR), the NHS Economic Evaluation Database (NHS EED), and the Health Economic Evaluations Database (HEED). Additional references were identified through reviewing manufacturer and sponsor submissions made to NICE, the bibliographies of retrieved articles, and conferences proceedings on the Internet. Only randomised controlled trials (RCTs) or non-randomised controlled trials with concurrent controls and full economic evaluations were considered for inclusion. Only studies that evaluated some sort of debriding method or a specialised wound care clinic (a nurse with specialist training in wound care; care being provided by a multidisciplinary team; a fast track referral system to other professions (e.g. dermatologist); or access to the latest health technology) were included in the review. Studies had to include participants with surgical wounds healing by secondary intention (e.g. cavity wounds, the consequences of wound dehiscence and abscesses) and report an objective measure of wound healing. Data were extracted by one reviewer and checked by a second. Quality assessment was conducted independently by two reviewers. Disagreements were resolved by consensus and, when necessary, by recourse to a third reviewer. The primary outcomes of interest were wound healing and cost. Results of data extraction and quality assessment were presented in structured tables and also as a narrative summary. In addition, where feasible, the results of individual studies were presented as forest plots. Studies were grouped according to the type of wound, debriding method and outcome measure used. RESULTS - CLINICAL EFFECTIVENESS: Seventeen trials met the inclusion criteria, all of which used the autolytic method of debridement. No studies were found that investigated sharp/surgical, biosurgical, mechanical, chemical or enzymatic debridement in the treatment of surgical wounds healing by secondary intention. No studies were found which investigated specialised wound care clinics that included the provision of care within a clinical setting (based in either primary or secondary care). The type of surgical wounds investigated by studies included in the review were those that had broken down postoperatively, perineal wounds resulting from proctolectomy or rectal excision, and those left open after pilonidal sinus excision or abscess incision, or wounds following a laparotomy. Four additional studies investigated treatment of postoperative wounds from toenail avulsions. (ABSTRACT TRUNCATED) PMID- 11399238 TI - A rapid and systematic review of the clinical effectiveness and cost effectiveness of orlistat in the management of obesity. AB - BACKGROUND: The prevalence of obesity in developed societies is increasing. Obesity is associated with an increased risk of co-morbidity, including cardiovascular disease and diabetes. Following the withdrawal of fenfluramine and dexfenfluramine, interest has focused on a novel anti-obesity drug orlistat. OBJECTIVE: To systematically assess the clinical effectiveness and cost effectiveness of orlistat in the management of obesity. METHODS - SEARCH STRATEGY: Nineteen electronic databases were searched from inception to June 2000. Additionally, Internet searches were carried out, bibliographies of retrieved articles were examined and submissions were received from the manufacturer of orlistat. METHODS - INCLUSION AND EXCLUSION CRITERIA: Randomised controlled trials (RCTs) evaluating the effectiveness of orlistat used for weight loss or maintenance of weight loss in overweight or obese patients were eligible for inclusion. Primary outcome measures were changes in body weight, fat content or fat distribution. Secondary outcomes were changes in obesity-related risk factor profiles, such as lipid levels, indicators of glycaemic control and blood pressure. Studies recruiting people with eating disorders such as anorexia nervosa and bulimia nervosa were excluded. METHODS - PROCESS OF STUDY SELECTION: Assessment of titles and abstracts was performed independently by two reviewers. If either reviewer considered a reference to be relevant, the full paper was retrieved. Full papers were assessed against the review selection criteria by two independent reviewers, and disagreements were resolved through discussion. METHODS - DATA EXTRACTION: Data were extracted by one reviewer into structured summary tables and checked by a second reviewer. Any disagreements about data were resolved by discussion. METHODS - QUALITY ASSESSMENT: Each included trial was assessed against a comprehensive checklist for methodological quality. Quality assessment was performed independently by two reviewers with disagreements resolved by discussion. METHODS - METHODS OF ANALYSIS/SYNTHESIS: This report is a narrative summary, with results grouped according to study endpoint. Statistical pooling was undertaken in groups of trials that were considered to be sufficiently similar. METHODS - ESTIMATION OF QUALITY OF LIFE, COSTS AND COST-EFFECTIVENESS AND/OR COST PER QUALITY-ADJUSTED LIFE-YEAR: Relevant economic evaluations were identified from the search strategy described above. Assessment of methodological quality was undertaken using principles outlined in published guidelines. METHODS - COMPANY SUBMISSIONS: Data from company submissions were subject to the same selection and appraisal processes as other studies considered for inclusion in the review, except that only RCTs with a duration of at least 1 year were selected. RESULTS - RESULTS OF THE SEARCH STRATEGY: Fourteen RCTs (including three company submissions) and two economic evaluations (including one company submission) were included in the review. RESULTS - RESULTS OF THE QUALITY ASSESSMENT: Methodological quality of trials was moderate to good. The main problems were lack of detail on methods used to produce true randomisation, small sample sizes in some cases and failure to use intention-to-treat analysis. It is likely that maintenance of blinding was difficult due to adverse effects associated with the study medication. RESULTS - EVIDENCE OF CLINICAL EFFECTIVENESS AND COST-EFFECTIVENESS: Most of the trials showed greater weight loss and better weight maintenance with orlistat compared to placebo at all endpoints (statistically significant differences for both outcomes). Orlistat 120 mg three times daily was the optimum regimen in terms of weight loss. Most trials showed significant improvement in at least some lipid concentration parameters, and, in three RCTs, orlistat produced statistically significant reductions in blood pressure relative to placebo. In obese patients with type 2 diabetes, orlistat resulted in a significantly greater weight loss at 1 year compared with placebo, and some parameters of glycaemic control and lipid concentration also showed significantly greater improvements compared with placebo. The incidence of gastrointestinal adverse events was consistently higher in orlistat groups compared with placebo, and orlistat use was associated with lower serum levels of fat-soluble vitamins. The cost per quality-adjusted life year for orlistat was 45,881 UK pounds. CONCLUSIONS - IMPLICATIONS FOR CLINICAL PRACTICE: Although many trials have demonstrated statistically significant differences between groups in terms of weight loss in favour of orlistat versus placebo, the differences may not always be of clinical significance. The clinical significance of between-group differences for secondary outcomes may also be debatable. Possible adverse effects should be taken into account when prescribing orlistat, particularly gastrointestinal effects. (ABSTRACT TRUNCATED) PMID- 11399239 TI - Multiple interactions determine cellular electrical processes in the multicellular tissue. PMID- 11399240 TI - Help from within: cardioprotective properties of hepatocyte growth factor. PMID- 11399241 TI - Perinatal vascular development. PMID- 11399242 TI - The importance of calcium in interpretation of NaK-ATPase isoform function in the mouse heart. PMID- 11399243 TI - Myocardial ischemia and infarction: growth of ideas. AB - This report reviews the author's involvement in the growth of ideas and basic concepts in myocardial ischemia resulting in the histological changes of myocardial infarction. Concepts arising from the study of myocardial substrate utilization, activation of the inducible form of nitric oxide synthase and production of prostacyclin and thromboxane in the infarcted heart are presented. New approaches are discussed dealing with the effects of nonsteroidal anti inflammatory drugs on myocardial production of nitric oxide and prostanoids, and with the relevance of the inducible form of cyclooxygenase. The review also records a number of significant similarities between angiogenesis in the ischemic heart and some cancers. Angiogenesis in both instances originates from inflammatory reactions, illustrating how different tissues and organs such as ischemic heart muscle and cancer react to similar pathological stimuli in an identical manner. This multifocal approach opens new concepts on myocardial ischemia and cancer. PMID- 11399244 TI - Influence of L-carnitine and its derivatives on myocardial metabolism and function in ischemic heart disease and during cardiopulmonary bypass. AB - Carnitine and its derivatives have recently been shown to protect cardiac metabolism and function in ischemic heart disease and other clinical conditions of myocardial ischemia. Potential mechanisms of this effect include an increase in glucose metabolism, a reduction of toxic effects of long-chain acyl-CoA and acyl-carnitine in myocytes, an increase in coronary blood flow and anti arrhythmic effect. It has also been shown that propionyl-L-carnitine which penetrates faster than carnitine into myocytes is effective in inhibiting production of free radicals. Beneficial effects of carnitine supplementation have been demonstrated under a variety of clinical conditions such as acute cardiac ischemia, during extracorporeal circulation, in carnitine-dependent cardiomyopathy as well as in patients with chronic circulatory failure and in cardiogenic shock. However, further studies are required before carnitine administration could be recommended as a routine procedure in ischemic heart disease or before cardiopulmonary bypass. PMID- 11399245 TI - Effects of cell-to-cell uncoupling and catecholamines on Purkinje and ventricular action potentials: implications for phase-1b arrhythmias. AB - OBJECTIVE: The delayed phase of ventricular arrhythmias during acute ischemia (phase-1b arrhythmia) is associated with depletion of catecholamines and cell-to cell uncoupling between depressed depolarized intramural ischemic region and surviving cells in subepicardium and subendocardium. In the present study we determined the effects of uncoupling and catecholamines on development of proarrhythmic afterdepolarizations. METHODS: Depressed depolarized ischemic region was simulated by a passive electronic circuit with a potential of -73, 53, -33 or -13 mV. Using patch-clamp methodology, single sheep Purkinje and ventricular cells were coupled to the simulated ischemic region via a variable conductance. By varying coupling conductance, we were able to selectively study the effects of various degrees of uncoupling. RESULTS: At strong coupling, cells were inexcitable and depolarized to potentials near those of the simulated ischemic region. Excitability, action potential duration and resting potential increased with progressive uncoupling. In a critical range of uncoupling, ventricular and "high-plateau" Purkinje cells developed early afterdepolarizations when the potential of the simulated ischemic region was -13 mV. Norepinephrine (1 microM) frequently induced early and delayed afterdepolarizations in both ventricular and Purkinje cells, but these afterdepolarizations were only present during uncoupling when the potential of the simulated ischemic region was -33 mV or more positive. CONCLUSIONS: In a critical range of uncoupling, afterdepolarizations were present when the potential of the simulated ischemic region was -33 or -13 mV, suggesting that triggered activity plays a role in phase-1b arrhythmias when surviving layers uncouple from a highly depolarized intramural ischemic region. PMID- 11399246 TI - A potential cardioprotective role of hepatocyte growth factor in myocardial infarction in rats. AB - OBJECTIVE: Cardiotrophic growth factors with anti-cell death actions on cardiac myocytes have gained attention for treatment of patients with myocardial infarction. Hepatocyte growth factor (HGF) plays a role in tissue repair and protection from injuries, however, the physiological role of HGF in the myocardium has not been well defined. We asked if HGF would afford to the infarcted myocardium. METHODS AND RESULTS: Mature cardiac myocytes prepared from adult rats expressed barely detectable levels of the c-Met/HGF receptor, however, c-Met receptor expression increased during cultivation, which meant that cardiac myocytes are potential targets of HGF. Addition of hydrogen peroxide remarkably decreased the number of viable mature cardiac myocytes in primary culture, whereas treatment with HGF enhanced survival of the cells subjected to the oxidant stress. Although very low levels of c-Met/HGF receptor and HGF mRNA expression were seen in normal rat hearts, both c-Met/HGF receptor and HGF mRNA levels rapidly increased to much higher levels than normal, when the rats were subjected to myocardial infarction. Immunohistochemical analysis of the c-Met receptor indicated that this receptor was expressed in cardiomyocytes localized in the border regions of the viable myocardium and in non-infarcted regions following myocardial infarction. CONCLUSION: The c-Met/HGF receptor is induced in cardiomyocytes following myocardial infarction and HGF exhibits protective effect on cardiomyocytes against oxidative stress. Our working hypothesis is that HGF may afford myocardial protection from myocardial infarction. PMID- 11399247 TI - Nitric oxide does not modulate the hyperpolarization-activated current, I(f), in ventricular myocytes from spontaneously hypertensive rats. AB - OBJECTIVE: : In sinoatrial (SA) node cells, nitric oxide (NO) exerts a dual effect on the hyperpolarization-activated current, I(f), i.e. in basal conditions NO enhances I(f) whereas in the presence of beta-adrenergic stimulation it decreases it. Recent studies have shown that I(f) is present in ventricular myocytes from hypertrophied or failing hearts where it may promote abnormal automaticity. Since these pathological conditions are associated with increased sympathetic tone and upregulation of myocardial NO production, we set out to investigate whether I(f) is similarly modulated by NO in hypertrophied ventricular myocytes. METHODS: Left ventricular myocytes were isolated from 18-20 month-old spontaneously hypertensive rats (SHRs). Membrane current was measured under whole-cell or amphotericin-perforated patch-clamp conditions, at 35 degrees C. RESULTS: Application of diethylamine-NO (DEA-NO, 1-100 microM) did not alter the amplitude or voltage dependence of activation of I(f) under basal conditions (half-activation voltage, V(h): control -82.9+/-2.6, DEA-NO -84.0+/-2.6 mV). Similarly, I(f) was not affected by the inhibition of endogenous NO production (L NMMA, 500 microM) or guanylate cyclase (ODQ, 10 microM). Forskolin (10 microM) or isoprenaline (100 nM) elicited a positive shift in V(h) but subsequent application of DEA-NO did not further affect the properties of I(f). CONCLUSIONS: Our results show that, unlike in SA node cells, in SHR ventricular myocytes basal and adrenergically stimulated I(f) is not modulated by exogenous NO or by constitutive NO or cGMP production. PMID- 11399248 TI - Ionic basis for membrane potential changes induced by hypoosmotic stress in guinea-pig ventricular myocytes. AB - OBJECTIVE: Causal relation between changes in action potentials and activation of several ionic currents during hypoosmotic challenge was investigated. METHODS: We recorded changes in membrane potentials and currents during hypotonic stress in guinea-pig ventricular myocytes using whole-cell patch-clamp technique. RESULTS: Exposure of ventricular myocytes to hypotonic solution (0.6 T) caused initial prolongation ( approximately 107% of control) of action potential duration at 90% repolarization (APD(90)) in 65% of examined myocytes. Later shortening (approximately 75% of control) of APD(90) and depolarization of resting potential (RP) (approximately 4 mV) developed in all cells. Initial prolongation of APD(90) in hypotonic solution was mainly caused by transient activation of Gd(3+) sensitive non-selective cation (NSC) current. Late changes after approximately 180 s in hypotonic solution were sustained increase in slow component of delayed rectifier K(+) current (I(Ks)) in all cells, and activation of I(Clswell) in 40% of cells. Prevention of APD(90) shortening by chromanol, a selective blocker of I(Ks), was seen in about 40% of myocytes due to short APD in our experimental conditions. Application of 1 mM anthracene-9-carboxylic acid (9-AC) partially inhibited APD shortening in three of seven cells. Depolarization of RP was unaffected by the above-mentioned drugs, but was dependent on [K(+)](o). CONCLUSIONS: Initial prolongation followed by later shortening of APD in hypotonic solution are mostly caused by different sequences of NSC, I(Ks) and I(Clswell) currents activation. Depolarization of RP in hypotonic solution is probably due to dilution of subsarcolemmal K(+) concentration and/or change in permeability ratio for Na(+) and K(+). PMID- 11399249 TI - Hyperactivity and altered mRNA isoform expression of the Cl(-)/HCO(3)(-) anion exchanger in the hypertrophied myocardium. AB - OBJECTIVE: The aim was to examine the regulation of the cardiac Na(+)-independent Cl(-)/HCO(3)(-) exchanger (AE) mRNA isoform expression in association to the enhanced AE activity in the hypertrophied myocardium of spontaneously hypertensive rats (SHR). METHODS: AE activity was determined by the initial rates of the pH(i) recovery from imposed intracellular alkalinization (forward mode of exchange) and the pH(i) rise induced by Cl(-) removal (reverse mode). Net HCO(3)( ) (J(HCO(3)(-))) efflux and influx were respectively determined. AE mRNA isoforms were analyzed by Northern blot with specific probes to detect AE1, AE2 and AE3 mRNAs. RESULTS: Initial J(HCO(3)(-)) efflux after imposed alkaline load (pH(i) congruent with 7.5) was higher in SHR than in normotensive WKY rats (3.01+/-0.33, n=7, vs. 0.64+/-0.29 mM/min, n=5, P<0.05). J(HCO(3)(-)) influx induced by Cl(-) deprivation was also increased in SHR, 4.24+/-0.56 mM/min (n=10) versus 2.31+/ 0.26 (n=10, P<0.05) in WKY. In arbitrary units, the 4.1-kb AE1 mRNA decreased in SHR (0.15+/-0.01, n=7) compared to WKY (0.29+/-0.06, n=7, P<0.05), whereas the 3.6-kb mRNA did not change. AE2 mRNAs were similarly expressed in WKY and SHR. Cardiac specific AE3 (cAE3) mRNA decreased in SHR, 1.10+/-0.16 arbitrary units (n=8) versus 1.79+/-0.24, (n=8, P<0.05) in WKY. Full length AE3 (flAE3) mRNA increased from 0.69+/-0.06 (WKY, n=8) to 1.25+/-0.19 arbitrary units in SHR (n=8, P<0.05). CONCLUSIONS: The increase in flAE3 mRNA expression in cardiac tissue from the SHR is an adaptive change of the hypertrophied myocardium that might be in connection with the increased activity of the AE. PMID- 11399250 TI - PKA-dependent phosphorylation of cardiac myosin binding protein C in transgenic mice. AB - OBJECTIVE: To investigate the physiological role of cAMP-dependent protein kinase A (PKA)-mediated, cardiac myosin binding protein C (MyBP-C) phosphorylation. METHODS: A cardiac MyBP-C cDNA lacking nine amino acids, which contained a phosphorylation site, was made, and subsequently used to generate multiple lines of transgenic mice. Upon confirming that a partial replacement of endogenous protein with transgenic protein occurred, the biochemical and physiological consequences were studied. PKA-dependent phosphorylation assays were used to estimate the phosphorylation states of major cardiac PKA substrates. Myofibril Mg ATPase activities were also measured. Isolated working heart and whole animal exercise studies were used to measure the physiological changes. RESULTS: Transgenic mice displayed a compensatory response, with PKA-mediated phosphorylation of both troponin I and phospholamban showing significant increases. The remaining endogenous cardiac MyBP-C also showed increased phosphorylation levels. Maximal Mg(2+)-ATPase activity was increased. Significant functional changes at both the whole organ and whole animal levels also occurred. Parameters reflecting cardiac contractility and relaxation increased about 22 and 25%, respectively, in the mutant relative to wild type mice (n=5, P<0.001). In young adults the capacity for stress exercise, quantitated using an exercise treadmill regimen, was substantially enhanced (n=6, P<0.01). CONCLUSIONS: Cardiac MyBP-C phosphorylation plays an important physiological role and that the protein's degree of phosphorylation is coordinated with the phosphorylation levels of other proteins within the contractile apparatus. PMID- 11399251 TI - Postischemic myocardial recovery and oxidative stress status of vitamin C deficient rat hearts. AB - OBJECTIVE: To investigate the role of vitamin C tissue content as a protective agent during myocardial ischemia-reperfusion injury, we have evaluated the postischemic functional recovery and free radical release of osteogenic disorder Shionogi (ODS) inherently scorbutic rat hearts and compared them to healthy Wistar rat hearts. METHODS: Isolated perfused hearts of ODS or Wistar rats underwent 30 min of a global total normothermic ischemia followed by 30 min of reperfusion. The lipid-soluble spin trap alpha-phenyl N-tert-butylnitrone (3 mM) was perfused upstream of the coronary bed. Functional parameters were recorded and samples of coronary effluents were analysed using electron spin resonance spectroscopy to characterise and quantify the amount of radical species released. RESULTS: From the onset of reperfusion, a large and long-lasting release of alkyl/alkoxyl radicals was detected, with a peak value of 29.0+/-3.2 nM obtained after 13 min, which was associated with a persistent contractile dysfunction. However, ODS rat hearts showed a higher myocardial recovery with lower left ventricular end diastolic pressure (44.34+/-1.74 vs. 55.03+/-1.57 mmHg for Wistar), higher recovery of rate pressure product (12.3+/-1.4 vs. 1.9+/-1.7x10(3) mmHg beats/min for Wistar) and shorter duration of contractile abnormalities during reperfusion (3.7+/-1.0 vs. 20.8+/-5.3 min for Wistar). Moreover, free radical release was identical in ODS rat hearts as compared to control Wistar rats. Ascorbic acid tissue content was significantly altered in ODS rats (31.9+/ 3.3 vs. 591.0+/-54.9 mmol/g of tissue for Wistar) but superoxide dismutases, glutathion peroxidases and inducible heat shock protein 70 genes were up regulated. CONCLUSIONS: This study shows that ascorbic-acid-deficient ODS rat hearts are more resistant to an ischemic insult than control Wistar rats, probably through the development of alternative protective defences, like the induction of heat shock proteins. These paradoxical results raise the question of the relative importance of each endogenous antioxidant in the cardiac resistance to ischemia-reperfusion injury. PMID- 11399252 TI - Short-acting calcium antagonist clevidipine protects against reperfusion injury via local nitric oxide-related mechanisms in the jeopardised myocardium. AB - BACKGROUND: Calcium antagonists may, in addition to their classical actions, release nitric oxide (NO) from coronary arteries. The aim of this study was to elucidate the possible interaction between the cardioprotective effect of a short acting calcium antagonist and NO during myocardial ischaemia and reperfusion. METHODS: Anaesthetised pigs were subjected to 45 min ligation of the left anterior descending coronary artery (LAD) followed by 4 h of reperfusion. Five groups were given vehicle (n=9), clevidipine (n=8), the NO synthase inhibitor L NMMA (n=6), clevidipine in combination with L-NMMA (n=6) or clevidipine in combination with L-NMMA and NO precursor L-arginine (n=6) into the LAD during the last 10 min of ischaemia and the first 5 min of reperfusion. RESULTS: There were no significant differences in LAD blood flow, mean arterial pressure, rate pressure product or dP/dt between the groups before ischaemia or during reperfusion. The infarct size (IS) was 86+/-2% of the area at risk in the vehicle group. Clevidipine reduced the IS to 59+/-3% (P<0.001). When clevidipine was administered together with L-NMMA, the protective effect of clevidipine was abolished (IS, 87+/-3%; P<0.001 vs. clevidipine), whereas addition of L-arginine restored its cardioprotective effect (IS 60+/-3%; P<0.001 vs. vehicle). L-NMMA did not affect IS per se (88+/-5%). Endothelium-dependent coronary vasodilation induced by substance P was significantly larger in the clevidipine group than in the other groups. CONCLUSION: Local administration of a calcium antagonist during the late ischaemia and early reperfusion reduces IS and preserves coronary endothelial function. The cardioprotective effect of clevidipine is suggested to be dependent on maintained local bioavailability of NO. PMID- 11399253 TI - Post-ischemic PKC inhibition impairs myocardial calcium handling and increases contractile protein calcium sensitivity. AB - OBJECTIVE: Protein kinase C (PKC) activation impairs contractility in the normal heart but is protective during myocardial ischemia. We hypothesized that PKC remains activated post-ischemia and modulates myocardial excitation-contraction coupling during early reperfusion. METHODS: Langendorff-perfused rabbit hearts where subjected to 25 min unmodified ischemia and 30 min reperfusion. Total PKC activity was measured, and the intracellular translocation pattern of PKC-alpha, delta, -epsilon, and -eta assessed by immunohistochemistry and fractionated Western immunoblotting. The PKC-inhibitors chelerythrine and GF109203X were added during reperfusion and also given to non-ischemic hearts. Measurements included left ventricular function, intracellular calcium handling measured by Rhod-2 spectrofluorometry, myofibrillar calcium responsiveness in beating and tetanized hearts, and metabolic parameters. RESULTS: Total PKC activity was increased at end-ischemia and remained elevated after 30 min of reperfusion. The translocation pattern indicated PKC-epsilon as the main active isoform during reperfusion. Post ischemic PKC inhibition affected mainly diastolic relaxation, with lesser effect on contractility. Both PKC inhibitors increased the Ca(2+) responsiveness of the myofilaments as indicated by a leftward shift of the calcium-to-force relationship and increased maximum calcium activated tetanic pressure. Diastolic Ca(2+) removal was delayed and the post-ischemic [Ca(2+)](i) overload further exacerbated. Depressed systolic function was associated with a lower amplitude of [Ca(2+)](i) transients. CONCLUSION: PKC is activated during ischemia and remains activated during early reperfusion. Inhibition of PKC activity post-ischemia impairs functional recovery, delays diastolic [Ca(2+)](i) removal, and increases Ca(2+) sensitivity of the contractile apparatus, resulting in impaired diastolic relaxation. Thus, post-ischemic PKC activity may serve to restore post-ischemic Ca(2+) homeostasis and attenuate contractile protein calcium sensitivity during the period of post-ischemic [Ca(2+)](i) overload. PMID- 11399254 TI - Oxygen wastage of stunned myocardium in vivo is due to an increased oxygen cost of contractility and a decreased myofibrillar efficiency. AB - OBJECTIVE: We investigated whether an increased oxygen cost of contractility and/or a decreased myofibrillar efficiency contribute to oxygen wastage of stunned myocardium. Because Ca(2+)-sensitizers may increase myofibrillar Ca(2+) sensitivity without increasing cross-bridge cycling, we also investigated whether EMD 60263 restores myofibrillar efficiency and/or the oxygen cost of contractility. METHODS: Regional fiber stress and strain were calculated from mesomyocardially implanted ultrasound crystals and left ventricular pressure in anesthetized pigs (n=18). Regional myocardial oxygen consumption (MVO(2)) was measured before contractility (end-systolic elastance, E(es)) and total myofibrillar work (stress-strain area, SSA) were determined from stress-strain relationships. Atrial pacing at three heart rates and two doses of dobutamine were used to vary SSA and E(es), respectively. After stunning (two times 10-min ischemia followed by 30-min reperfusion), measurements were repeated following infusion of saline (n=8) or EMD 60263 (1.5 mg.kg(-1) i.v., n=10). Linear regression was performed using: MVO(2)=alpha.SSA+beta.E(es)+gamma.HR(-1) (alpha( 1), myofibrillar efficiency; beta, oxygen cost of contractility; and gamma, basal metabolism/min). RESULTS: Stunning decreased SSA by 57% and E(es) by 64%, without affecting MVO(2), while increasing alpha by 71% and beta by 134%, without affecting gamma. From the wasted oxygen, 72% was used for myofibrillar work and 18% for excitation-contraction coupling. EMD 60263 restored both alpha and beta. CONCLUSIONS: Oxygen wastage in stunning is predominantly caused by a decreased myofibrillar efficiency and to a lesser extent by an increased oxygen cost of contractility. Considering that EMD 60263 reversed both causes of oxygen wastage, it is most likely that this drug increases myofibrillar Ca(2+)-sensitivity without increasing myofibrillar cross-bridge cycling. PMID- 11399255 TI - Combined angiotensin converting enzyme inhibition and angiotensin AT(1) receptor blockade up-regulates myocardial AT(2) receptors in remodeled myocardium post infarction. AB - OBJECTIVES: In an ovine model of left ventricular (LV) remodeling after transmural anteroapical myocardial infarction (MI), we have previously demonstrated that the combination of angiotensin converting enzyme (ACE) inhibition and AT(1) receptor blockade is more effective at limiting LV remodeling than either therapy alone. We hypothesized that the beneficial effect of combined therapy is due in part to upregulation of AT(2) receptor levels. METHODS: Two days after transmural anteroapical MI by coronary ligation, 16 sheep were randomized to losartan (50 mg/day), ramipril (10 mg/day), ramipril+losartan (combined therapy), or no therapy. At 8 weeks after MI, radioligand receptor assay were deployed with homogenates from regional LV tissues. RESULTS: We found that AT receptors in normal sheep myocardium are predominantly of the AT(2) receptor subtype. Binding studies of remodeled myocardium 8 weeks later showed that the apparent maximum binding (B(max)) was increased from 23 to 48 fmol/mg protein only in animals with combined therapy. The AT(2)/AT(1) proportion was increased significantly in animals with combined therapy compared to infarcted controls (18.0 vs. 5.17). CONCLUSIONS: These results indicate that AT(2) receptor expression increased significantly during LV remodeling with combined therapy but not with either therapy alone. In combination with prior work demonstrating the effectiveness of combined therapy in limiting LV remodeling, this study is consistent with the hypothesis that AT(2) receptors play a cardioprotective role in LV remodeling after MI. PMID- 11399256 TI - Perinatal development influences mechanisms of bradykinin-induced relaxations in pulmonary resistance and conduit arteries differently. AB - OBJECTIVE: As bradykinin (BYK) relaxes conduit (EPA) and resistance (RPA) pulmonary arteries from both perinatal and adult lungs, we investigated whether this vasodilator's relaxation-mechanisms were altered during perinatal development, differed between EPA and RPA and differed with other endothelium dependent vasodilators, acetyicholine (ACH) and substance P (SP). METHODS: Arteries from mature foetal (5 days), neonatal (approximately 5 min), newborn (60 84 h) and adult pigs (> or =6 months) were isolated, mounted for in vitro isometric force recording, activated with PGF(2alpha) (30 micromol/l) and relaxed with BYK (10 pmol/l-1 micromol/l), SP (10 pmol/l-0.1 micromol/l) or ACH (1 nmol/l 1 mmol/l). RESULTS: (i) BYK: L-NAME (100 micromol/l) attenuated relaxations in foetal EPA ( approximately 55%) but nearly abolished them in the adult ( approximately 80%). In RPA, L-NAME nearly abolished ( approximately 90%) relaxations in the foetus and this effect diminished progressively with age to approximately 20% in the adult. Indomethacin (IND, micromol/l) attenuated relaxations in neonatal (approximately 25%), new-born and adult EPA (both approximately 45%). Together, L-NAME and IND abolished relaxations in all EPA and in neonatal RPA but not in older RPA. SKF525a (100 micromol/l) attenuated relaxations in foetal RPA ( approximately 4%), diminishing in the adult RPA to approximately 10%. Together, SKF52Sa and L-NAME largely abolished relaxations in postnatal RPA (approximately 80%). Activation with K(+)=125 mmol/l attenuated relaxations in adult EPA (approximately 80%), foetal RPA ( approximately 45%) and neonatal RPA (approximately 75%) and abolished relaxations in RPA from older ages. (ii) ACH: L-NAME abolished relaxations in new-born EPA and RPA. In adult EPA, combined L-NAME and IND moderately attenuated relaxations. (iii) SP: Combined application of L-NAME and IND attenuated relaxations to a similar degree in new-born and adult EPA and RPA. CONCLUSIONS: In postnatal EPA, BYK-relaxations depend completely on prostaglandin- and NO-synthesis whereas those to SP (at all ages) and ACH (in the adult) do not. In RPA, BYK-relaxations develop from being completely dependant on the sole release of NO (foetus) to being almost completely independent of it (adult), a situation mimicked partially by SP but not by ACH, which, in new-born RPA is completely dependent on NO. BYK-relaxations in postnatal RPA depend on the release of a hyperpolarising factor generated through an SKF525a-sensitive pathway in conjunction with NO. The mechanisms of endothelium-dependent BYK-relaxations in the pulmonary vascular bed undergo diverging alterations, depending on the stage of development and arterial size/function. These changes are specific for BYK as they differ from those obtained from ACH or SP. PMID- 11399257 TI - Combined effect of retinoic acid and basic FGF on PAI-1 gene expression in vascular smooth muscle cells. AB - OBJECTIVE: Aberrant regulation of the synthesis and degradation of the extracellular matrix (ECM) is associated with the pathophysiology of vascular disease. Plasminogen activator inhibitor-1 (PAI-1) plays a crucial role in regulating the quantity and composition of ECM. However, regulatory mechanisms underlying the expression of the PAI-1 gene remain unclear. We examined the effects of all-trans-retinoic acid (atRA), either alone or in combination with mitogenic growth factor, basic fibroblast growth factor (bFGF), on the PAI-1 expression in cultured vascular smooth muscle cells (SMCs). METHODS: Cultures of the rabbit vascular smooth muscle cell line C2/2 were used to study the effects of atRA and bFGF separately or together. RESULTS: Treatment of vascular SMCs with atRA in combination with bFGF resulted in an additional increase in PAI-1 expression both at the mRNA and protein levels. In contrast, tissue-type plasminogen activator, urokinase-type plasminogen activator and tissue factor mRNA levels were only minimally affected. The all-trans-RA- and bFGF-mediated increases in PAI-1 mRNA levels were markedly attenuated by the tyrosine kinase inhibitor genistein, but not by MEK1 or p38MAP kinase inhibitors. The rate of decrease in PAI-1 mRNA levels after actinomycin D treatment was not affected by atRA and bFGF. Transient transfection of the PAI-1 promoter-luciferase reporter gene, which contains 967 bp of the 5'-flanking region of the human PAI-1 gene, revealed that atRA and bFGF additionally increased transcription from this promoter. Progressive 5'-deletion revealed that the promoter region required for such an effect lies between -967 and -260, which contains no canonical sequence for the RA-response element. In agreement with the role of PAI-1 in the inhibition of fibrinolytic activity which stimulates ECM degradation, cell migration was inhibited by treatment with atRA and bFGF. CONCLUSIONS: These results indicate that atRA and bFGF can function in a combined fashion and induce PAI-1 synthesis in vascular SMCs, and suggest a role for these two compounds in regulating SMC migration. PMID- 11399258 TI - Expression of ryanodine receptor type 3 and TRP channels in endothelial cells: comparison of in situ and cultured human endothelial cells. AB - OBJECTIVE: Ca(2+) mobilization plays an important role in endothelial function by stimulating Ca(2+)-dependent synthesis of vasodilating factors. In addition to inositol-1,4,5-trisphosphate (InsP(3)) mediated Ca(2+) mobilization, Ca(2+) release from ryanodine-sensitive pools and Ca(2+)-influx through TRP channels have been suggested to be important in endothelial Ca(2+)-signaling. However, the function and molecular identity of TRP channels and ryanodine receptors in human endothelium in situ are still elusive. We hypothesized that expression of ryanodine-receptors (RyR) and TRP channels differs between human endothelium in situ and in cultured cells. METHODS: By combining single-cell RT-PCR and patch clamp techniques, expression of RyR and TRP channels was determined in situ in endothelial cells of human mesenteric artery (HMAECs) obtained from patients undergoing bowel resection and in the endothelial cell line EA.hy926. RESULTS: At the single cell level, expression of RyR 3 was detected in 25 and 5% of HMAECs and EA.hy926 samples, respectively. Expression of the RyR 1 and 2 was not detected in either HMAECs or EA.hy926. In patch-clamp experiments in HMAECs, applications of caffeine (0.5 mM) induced sustained hyperpolarization mediated by activation of Ca(2+)-activated K channels. In EA.hy926, caffeine-induced hyperpolarization was not detected. Single HMAECs expressed the TRP genes, TRP1 and TRP3, but not TRP 4 and 6. The TRP1 was the predominantly expressed TRP gene in HMAECs in situ whereas TRP3 expression was rarely detected. EA.hy926 expressed only TRP1. In patch clamp experiments in HMAECs, Ca(2+)-store depletion activated non-selective cation currents leading to Ca(2+) entry. CONCLUSIONS: Our findings suggest that, in addition to InsP(3) mediated Ca(2+) release, Ca(2+) release from ryanodine-sensitive stores mediated by RyR3 and Ca(2+) entry through TRP1 might represent important components of endothelial Ca(2+) signaling in situ and thereby of endothelial function in intact human blood vessels. PMID- 11399259 TI - Long-term inhibition of Rho-kinase induces a regression of arteriosclerotic coronary lesions in a porcine model in vivo. AB - OBJECTIVE: We recently demonstrated that Rho-kinase/ROK/ROCK is functionally upregulated at the arteriosclerotic coronary lesions and plays a key role for coronary vasospastic responses in our porcine model with interleukin (IL)-1beta. In the present study, we tested our hypothesis that Rho-kinase is involved in the pathogenesis of coronary arteriosclerosis per se in our porcine model. METHODS: Segments of the left porcine coronary artery were chronically treated from the adventitia with IL-1beta. Two weeks after the procedure, coronary stenotic lesions with constrictive remodeling and vasospastic response to serotonin were noted at the IL-1beta-treated site, as previously reported. Then, animals were randomly divided into two groups; one group was treated with fasudil for 8 weeks followed by 1 or 4 weeks of washout period and another group served as a control. After oral absorption, fasudil is metabolized to hydroxyfasudil that is a specific inhibitor of Rho-kinase. RESULTS: In the fasudil group, coronary stenosis and vasospastic response were progressively reduced in vivo, while the coronary hyperreactivity was abolished both in vivo and in vitro. Furthermore, Western blot analysis showed that in the fasudil group, the Rho-kinase activity (as evaluated by the extent of phosphorylation of myosin binding subunit of myosin phosphatase, one of the major substrates of Rho-kinase) was significantly reduced, while histological examination demonstrated a marked regression of the coronary constrictive remodeling. CONCLUSIONS: These results indicate that Rho kinase is substantially involved in constrictive remodeling and vasospastic activity of the arteriosclerotic coronary artery, both of which could be reversed by long-term inhibition of the molecule in vivo. Thus, Rho-kinase may be regarded as a novel therapeutic target for arteriosclerotic vascular disease. PMID- 11399260 TI - Mechanical properties and structure of carotid arteries in mice lacking desmin. AB - OBJECTIVE: Our aim was to determine in desmin homozygous mutant mice the viscoelastic properties, the mechanical strength and the structure of the carotid artery. METHODS: To assess the viscoelastic properties of large arteries, we have performed an in vivo analysis of the diameter-, and distensibility-pressure curves of the common carotid artery (CCA) in homozygous (Des -/-), heterozygous (Des +/-) and wild-type (Des +/+) mice. To evaluate the mechanical strength, we have measured the in vitro intraluminal pressure producing the rupture of the carotid artery wall. The structure analysis of the arterial wall was based on histology and electronic microscopy. RESULTS: A lower distensibility and an increase of arterial wall viscosity were observed in Des -/- compared with Des +/+. Arterial thickness of Des -/- was similar to those of Des +/+, without changes in elastin and collagen contents. Electron microscopy revealed that the perimeter of cellular fingerlike-projections was smaller in Des -/-, indicating that the cells have lost part of their connections to the extracellular matrix. The rupture pressure was significantly lower in Des -/- (1500+/-200 mmHg) compared with Des +/+ (2100+/-80 mmHg) indicating a lower mechanical strength of the vascular wall. No significant difference was found between Des +/- and Des +/+. CONCLUSION: The desmin is essential to maintain proper viscoelastic properties, structure and mechanical strength of the vascular wall. PMID- 11399261 TI - Site-directed mutagenesis of the 5-HT1B receptor increases the affinity of 5-HT for the agonist low-affinity conformation and reduces the intrinsic activity of 5 HT. AB - The antagonist radioligand [3H]GR125743 and the agonist radioligand [3H]5-HT were used to investigate the pharmacological characteristics of the G protein uncoupled agonist low-affinity and G protein coupled agonist high-affinity conformations of the wild-type and mutant human 5-hydroxytryptamine 1B (5-HT1B) receptors. We found that substitution of phenylalanine 185 in transmembrane region IV by alanine or methionine resulted in a reduced number of receptors in the coupled conformation, as well as a reduced affinity of 5-HT for the uncoupled conformation. In contrast, substitution of phenylalanine 331 in transmembrane region VI by alanine increased the affinity of 5-HT for the uncoupled conformation 11-fold thus reducing the agonist low-affinity to agonist high affinity (K(il)/K(ih)) ratio 5-fold. This reduced ratio was correlated with a significantly reduced intrinsic activity of 5-HT previously determined by its ability to inhibit forskolin-stimulated cAMP production. In conclusion, these results show that single amino acid substitutions can selectively change the affinity of 5-HT for the G protein uncoupled conformation of the 5-HT1B receptor and alter the intrinsic activity of the ligand. PMID- 11399262 TI - The pharmacology of novel acetylcholinesterase inhibitors, (+/-)-huprines Y and X, on the Torpedo electric organ. AB - The effects of the tacrine-huperzine A hybrid acetylcholinesterase inhibitors, (+/-)-12-amino-3-chloro-9-methyl-6,7,10,11-tetrahydro-7,11 methanocycloocta[b]quinoline hydrochloride ((+/-)-huprine Y) and (+/-)-12-amino-3 chloro-9-ethyl-6,7,10,11-tetrahydro-7,11-methanocycloocta[b]quinoline hydrochloride ((+/-)-huprine X), were tested on spontaneous synaptic activity by measuring the amplitude, the rise time, the rate of rise, the half-width and the area or the electrical charge of the miniature endplate potentials (m.e.p.ps) recorded extracellularly on Torpedo electric organ fragments. (+/-)-Huprine Y and (+/-)-huprine X at a concentration of 500 nM increased all the m.e.p.p. variables analyzed. The effect of (+/-)-huprine Y was smaller than that of (+/-)-huprine X for all the variables except for the rate of rise where there was no significant difference. The effects of these drugs were also tested on nicotinic receptors by analyzing the currents elicited by acetylcholine (100 microM) in Xenopus laevis oocytes, transplanted with membranes from Torpedo electric organ. Both drugs inhibited the currents in a reversible manner, (+/-)-huprine Y (IC(50)=452 nM) being more effective than (+/-)-huprine X (IC(50)=4865 nM). The Hill coefficient was 0.5 for both drugs. The inhibition of the nicotinic receptor was voltage dependent and decreased at depolarizing potentials, and there was no significant difference in the effects between (+/-)-huprine Y and (+/-)-huprine X at concentrations near to their IC(50) values. At depolarizing potentials between 20 and +15 mV, these drugs did not have any detectable effect on the blockade of the nicotinic receptor. Both huprines increased the desensitization of the nicotinic receptors since the current closed quickly in the presence of the drugs, and there was no significant difference in this effect between (+/-) huprine Y (500 nM) and (+/-)-huprine X (5 microM). We conclude that (+/-)-huprine Y and (+/-)-huprine X increase the level of acetylcholine in the synaptic cleft more effectively than tacrine. The interaction of (+/-)-huprine X with nicotinic receptors is weaker than that of (+/-)-huprine Y, suggesting that (+/-)-huprine X would be more specific to maintain the extracellular acetylcholine concentration. PMID- 11399263 TI - Structural requirements of phenol derivatives for direct activation of chloride currents via GABA(A) receptors. AB - Propofol directly activates gamma-aminobutyric acid (GABA(A)) receptors in the absence of the natural agonist. This mechanism is supposed to contribute to its sedative-hypnotic actions. We studied the effects of seven structurally related phenol derivatives on chloride inward currents via rat alpha1beta2gamma2 GABA(A) receptors, heterologously expressed in HEK 293 cells in order to find structural determinants for this direct agonistic action. Only compounds with the phenolic hydroxyl attached directly to the benzene ring and with aliphatic substituents in ortho position to the phenolic hydroxyl activated chloride currents in the absence of GABA. Concentrations required for half-maximum effect were 980 microM for 2-methylphenol, 230 microM for 2,6-dimethylphenol, 200 microM for thymol, and 23 microM for propofol. Drug-induced chloride currents showed no desensitisation during the 2-s application. These results show that the position of the aliphatic substituents with respect to the phenolic hydroxyl group is the crucial structural feature for direct GABA(A) activation by phenol derivatives. PMID- 11399264 TI - Caspase-independent apoptosis induced by differentiation-inducing factor of Dicytostelium discoideum in INS-1 cells. AB - Differentiation-inducing factor (DIF) is a lipophilic hormone of Dicytostelium discoideum and has been shown to exert diverse effects in mammalian cells. We investigated the effect of DIF on cell viability in insulin-secreting INS-1 cells. DIF induced cell death in a dose-dependent manner. In DIF-treated cells, nuclear condensation and shrinkage of the cell body were observed. After 6 h of DIF treatment, cells became Tdt-mediated dUTP-biotin nick end-labeling-positive, and DNA ladder formation was detected, indicating that DIF induced apoptosis in these cells. DIF did not activate caspase-3, a key enzyme mediating apoptotic signals generated by various agents. Furthermore, DIF-induced cell death was not affected by Z-asp-2, 6-dichlorobenzoyloxymethylketone, a broad inhibitor of the caspases. As is the case in other types of cells, DIF increased cytoplasmic free calcium concentration in INS-1 cells. However, DIF-induced cell death was not affected by chelating intracellular free calcium by 1, 2-bis(2 aminoophenoxy)ethane-N, N, N, N-tetra acetic acid (BAPTA). These results indicate that DIF induces apoptosis in INS-1 cells by a mechanism independent of caspase 3. DIF-induced elevation of cytoplasmic calcium does not mediate the effect of DIF on cell death. PMID- 11399265 TI - Brain responses to acute withdrawal in phenobarbital-dependent rats. AB - Heat shock proteins (HSP) such as HO-1 and HSP27 have been implicated as functioning in a protective manner against oxidative and physical stress. The objective of the current study was to determine the role of HSPs in drug withdrawal stress induced in phenobarbital-dependent rats. Increased expression of HO-1 and HSP27 was observed in the hippocampus and the cerebral cortex of phenobarbital-withdrawn rats. Gene expression was measured by Northern and Western blot analyses and in situ hybridization. The induction of HO-1 mRNA was suppressed by the administration of the NMDA receptor antagonist, (+)-5-methyl 10,11-dihydro-5H-dibenzo (a,d) cyclohepten-5,10-imine (MK801). Despite significant upregulation of glutamatergic transmission, neuronal cell degeneration was not apparent. These findings suggest that the induction of HO-1 and HSP27 during withdrawal from phenobarbital dependence may play a role in protection against glutamate toxicity. PMID- 11399266 TI - In the formalin model of tonic nociceptive pain, 8-OH-DPAT produces 5-HT1A receptor-mediated, behaviorally specific analgesia. AB - The experiments examined antinociceptive and intrinsic behavioral effects induced by the prototypical 5-HT1A receptor agonist 8-OH-DPAT (8-hydroxy-2-[di-n propylamino] tetralin) in rats. 8-OH-DPAT (0.01-2.5 mg/kg, subcutaneous (s.c.)) reduced both the paw licking and paw elevation induced by (2.5%) formalin injection into the plantar surface of the right hindpaw; it also produced forepaw treading. All of these effects were completely blocked by pretreatment with WAY 100635 (N-(2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl)-N-(2-pyridinyl) cyclohexanecarboxamide trihydrochloride) (0.16 mg/kg, s.c.); prazosin (0.63 mg/kg, s.c.) inhibited forepaw treading, but not 8-OH-DPAT's action on paw elevation and paw licking. Repeated injection of 8-OH-DPAT (0.63 mg/kg, s.c.) twice daily for 4 days, markedly reduced 8-OH-DPAT's ability to produce forepaw treading, but exerted only little and inconsistent effects on its paw licking and paw elevation-inhibiting action. The data indicate that 8-OH-DPAT exerts an analgesic action in the formalin model of tonic nociceptive pain; this action is mediated by 5-HT(1A) receptors, and is not confounded by the productive sign (i.e., forepaw treading) of the 5-HT syndrome which 8-OH-DPAT also induces. PMID- 11399267 TI - Endomorphins 1 and 2, endogenous mu-opioid receptor agonists, impair passive avoidance learning in mice. AB - The effects of intracerebroventricular administration of endomorphin-1 and endomorphin-2, endogenous mu-opioid receptor agonists, on passive avoidance learning associated with long-term memory were investigated in mice. Endomorphin 1 (10 and 17.5 microg) and endomorphin-2 (17.5 microg) produced a significant decrease in step-down latency in a passive avoidance learning task. beta Funaltrexamine (5 microg) almost completely reversed the endomorphin-1 (17.5 microg)- and endomorphin-2 (17.5 microg)-induced shortening of step-down latency, although neither naltrindole (4 ng) nor nor-binaltorphimine (4 microg) produced any significant effects on the effects of endomorphins 1 and 2. These results suggest that endomorphins 1 and 2 impair long-term memory through the mediation of mu-opioid receptors in the brain. PMID- 11399268 TI - The influence of nitric oxide donors on the responses to nitrergic nerve stimulation in the mouse duodenum. AB - We investigated whether exogenous nitric oxide (NO) donors have a prejunctional and/or postjunctional inhibitory effect on the nitrergic responses and whether this inhibitory effect was mediated by NO itself and in part, by cyclic GMP in mouse duodenal strips. N(omega)-nitro-L-arginine inhibited relaxations induced by electrical field stimulation of nitrergic nerves, but not those with acidified NaNO2. Furthermore, 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one (ODQ) inhibited both types of relaxations while 2-amino-5,6-dihydro-6-methyl-4H-1,3-thiazine (AMT) and N-ethylmaleimide were ineffective. NO donors, nitroglycerin and sodium nitroprusside, inhibited relaxations induced by nitrergic nerve stimulation, but not those with acidified NaNO2. Hemoglobin, exogenous Cu(2+)/Zn(2+) superoxide dismutase, diethyldithiocarbamic acid and pyrogallol did not influence the relaxation with nitrergic nerve stimulation. However, hemoglobin, diethyldithiocarbamic acid, pyrogallol and diethyldithiocarbamic acid plus pyrogallol attenuated the inhibitory effect of NO donors on relaxation with nitrergic nerve stimulation, and exogenous superoxide dismutase potentiated this inhibitory effect. Moreover, nitrergic nerve-mediated relaxations were inhibited by 8-bromo-cyclic GMP, but not by 8-bromo-cyclic AMP. These results suggest that exogenous NO donors have a prejunctional inhibitory effect on the nerve-mediated nitrergic relaxation and that the inhibitory effects of nitroglycerin and sodium nitroprusside are NO-dependent, but not related to NO metabolites such as peroxynitrite or a nitrosothiol intermediate. However, a contribution of S nitrosothiol formed intracellularly cannot be entirely ruled out. Also, this prejunctional inhibition is mediated, at least in part, by the cyclic GMP, but not the cyclic AMP, pathway. PMID- 11399269 TI - Contribution of C-fibers to leucocyte recruitment in bronchoalveolar lavage fluid and pleural cavity in the rat. AB - The effect of neonatal capsaicin (8 methyl-N-vanillyl-6-nonenamide) treatment on the leucocyte infiltration into the airways and pleural cavity was investigated in rats actively sensitized with ovalbumin. The animals were neonatally injected with either capsaicin (50 mg/kg, s.c., 2nd day of life) or vehicle (10% ethanol and 10% Tween 80). At adult ages, the animals were actively sensitized with ovalbumin (200 microg, s.c.) and 14 days later they were intratracheally (or intrapleurally) challenged with ovalbumin. The substance P level in bronchoalveolar lavage fluid of the capsaicin group was reduced by >90% compared to control group (vehicle), confirming the efficacy of capsaicin treatment. In the capsaicin group, the number of neutrophils (but not of eosinophils and mononuclear cells) in bronchoalveolar lavage fluid of sensitized animals was significantly higher than the control group. Intrapleural injection of ovalbumin in sensitized rats caused a significant neutrophil influx at 6 h that was markedly increased in the capsaicin-pretreated animals compared to control group. The counts of eosinophils and mononuclear cells in the pleural exudates did not differ significantly between capsaicin and control groups. The increased levels of immunoglobulin (Ig)E, IgG1 and IgG2a anti-ovalbumin antibodies in serum of sensitized rats did not differ between capsaicin and control groups. In conclusion, the exacerbated pulmonary neutrophil recruitment caused by the capsaicin neonatal treatment is unrelated to increase in serum immunoglobulin antibodies, and suggests a protective role for C-fibers in attenuating the allergic neutrophil infiltration. PMID- 11399270 TI - Autoradiographic distribution of adrenomedullin receptors in the rat brain. AB - The autoradiographic distribution of putative brain adrenomedullin receptors was investigated using [125I]human adrenomedullin(13-52) as a new radioligand. Specific [125I]human adrenomedullin(13-52) binding sites were very discretely distributed in the rat brain with enrichment seen in the choroid plexus and linings of the third, fourth and lateral ventricles, basolateral amygdaloid nuclei, neural lobe of the pituitary gland, the trigeminal nerves and in the granular cell layer of the cerebellum. To our knowledge, this is the first report on the discrete localization of adrenomedullin receptors in the mammalian brain. PMID- 11399271 TI - Parental investment of male two-spotted goby, Gobiusculus flavescens (Fabricius). AB - The trade-off between parental care and feeding was studied in the male two spotted goby (Gobiusculus flavescens F.). Two temperatures, 8.5 degrees C and 13.0 degrees C, were used, with five replicates at each temperature, in order to determine whether temperature influenced parental behaviour. In each replicate, two males and four females were introduced to an aquarium, where the males chose between two nests and courted the females. In each replicate, one male spawned. After spawning, the males guarded the eggs until hatching. The guarding males' behaviour was recorded with a video camera twice a day (15 min each time), once before and once after they were fed. The male's condition (c-factor) was calculated at the start of the experiment and after egg hatching. The eggs were spawned in an artificial nest (half of a PVC tube), and attached to the nest in a single layer. The areas with eggs (representing brood size) were marked after spawning and the fry counted after hatching (which was used to calculate area hatched). Numbers of prey eaten (plankton) and number of aggressive encounters between the guarding male and the other fishes were recorded. Time spent in the nest and time used on fertilisation, fanning and cleaning were estimated and related to egg age, brood size, hatching success, temperature and food availability (no food or food).The results showed that feeding (expected to influence future reproduction) decreased and parental expenditure (current reproduction) increased, as the eggs developed (became closer to independence). Parental expenditure was significantly higher at 13.0 degrees C than at 8.5 degrees C, presumably due to higher oxygen demands by the eggs, and a greater risk of egg-infections. The c-factor of the males guarding eggs decreased over time, in contrast to the non-guarding males' c-factor. Guarding males' aggressiveness decreased as the eggs got older, but increased just before hatching. A possible explanation for this could be the decreasing intrusion by the non-guarding male and females caused by high aggressive behaviour by the guarding male early in the brood cycle. The exploitation of the nest (percentage of total nest area covered by eggs) seemed to determine the amount of parental expenditure and loss of condition, while brood size (area of eggs) had no effect. PMID- 11399272 TI - Mechanisms of interaction between macroalgae and scleractinians on a coral reef in Jamaica. AB - After several decades of disturbance, many coral reefs in the Caribbean are dominated by macroalgae. One process affecting this transition is coral macroalgal competition, yet few studies have addressed the mechanisms involved. In this study, we investigated competition between the tall and bushy macroalga Sargassum hystrix (J. Agardh) and the branching coral Porites porites (Pallas) on a shallow reef in Jamaica. Experiments were designed to expose coral branches to different treatments to test the role of shading and abrasion by Sargassum on coral growth and polyp expansion. Corals exposed to Sargassum grew significantly more slowly (80% reduction) than controls, but this effect was absent when corals were caged to prevent physical contact with macroalgae. Light levels were reduced in both the algal and cage treatments, but shading apparently had little effect on the growth of corals in cages. Short-term measurements of integrated net water flow did not detect variation among treatments. In algal-mimic treatments, where clear plastic strips could touch but not shade the corals, growth rates were 25% lower than controls, but this effect was not statistically significant. Thus, the growth of corals in contact with Sargassum was reduced by abrasion and, to a lesser extent, by factors unique to living macroalgae. Analysis of polyp expansion showed that polyps were more frequently retracted when corals were in contact with macroalgae or algal-mimics compared to controls or cage treatment; the frequency of polyp contraction was correlated positively with growth. Together, these results suggest that abrasion-mediated polyp retraction is one of the primary mechanisms of competition utilized by tall (ca. 17 cm) macroalgae against scleractinian corals. PMID- 11399273 TI - Feeding clusters and olfaction in the mangrove snail Terebralia palustris (Linnaeus) (Potamididae: Gastropoda). AB - Large numbers of the snail Terebralia palustris (Linnaeus) (Potamididae; Gastropoda) are frequently observed feeding in a cluster on a single fallen mangrove leaf, yet none are present on leaves nearby. Consequently, we investigated the food-finding ability of T. palustris in a Kenyan mangrove forest using field experiments. We estimated the attractive effect of different cues and analysed the paths (video-recorded) of snails when approaching a food-related odour. This intertidal snail can potentially use both air-borne and water-borne odours to locate food. T. palustris is attracted to conspecifics feeding on leaves, while intact leaves as well as non-feeding snails are not attractive. Moreover, the guiding stimulus appears to be compounds released when the leaves are damaged.T. palustris also seems able to discriminate between different food items; it is more strongly attracted to green mangrove leaves than senescent or fallen ones or mangrove propagules, probably because green leaves release a greater amount of attractive cues.Feeding snails thus recruit more snails to feed on the same leaf. The ecological implications of this behaviour are discussed: a large number of snails on the same leaf counteracts the ability of crabs to remove the leaf being eaten by the snails. PMID- 11399274 TI - Effects of the duration and timing of starvation during larval life on the metamorphosis and initial juvenile size of the polychaete Hydroides elegans (Haswell). AB - The larvae of the polychaete Hydroides elegans were exposed to different feeding schedules to evaluate the effects of starvation on metamorphosis and initial juvenile volume. The timing of starvation (early or late in development) and the total number of days of feeding (3, 5 or 8 days) at 24 degrees C had significant effects on the percentage of larvae that successfully completed metamorphosis and on the volume of the post-metamorphic juveniles. However, the duration of starvation (2, 4, 6 or 8 days) did not influence metamorphosis or juvenile volume. For larvae that had fed for 3 or 5 days, those that were starved at the beginning of development had a higher percentage of successful metamorphosis and produced larger juveniles compared with those starved after an initial 3-day feeding period. Larvae that had been allowed to feed for a total of 8 days were not affected by the timing of starvation. The percentage metamorphosis was lower in larvae that had fed for 3 days (16%), but was not significantly different between those that fed for 5 days (47%) compared to 8 days (47%). However, juvenile volume increased with the number of days of feeding from 0.95 nl after 3 days to 1.35 nl after 5 days, to 1.91 nl after 8 days of feeding. Discontinuous feeding reduced the proportion of larvae undergoing metamorphosis and juvenile volume compared to larvae continuously fed for only 5 days, but had no effect on larvae that fed for 8 days. The larvae of this extremely successful fouling species are remarkably tolerant of starvation and seem to require approximately 5 days of continuous feeding to achieve high levels of metamorphic success and large initial juvenile volume. PMID- 11399275 TI - Active habitat selection for sand by juvenile western king prawns, Melicertus latisulcatus (Kishinouye). AB - This paper presents the results of a series of habitat selection experiments aimed at determining if juvenile Melicertus latisulcatus generally occur on intertidal sand- and mud-flats as a result of active selection of unvegetated areas, or due to extrinsic factors (e.g. differential predation). In the laboratory, juvenile M. latisulcatus showed a clear preference for habitats containing sand irrespective of the presence or absence of predators. If sand was not available, artificial seagrass was chosen as a secondary preference but was avoided when sand alone was also present. Importantly, the combinations of habitats chosen for testing allowed us to determine that artificial seagrass provided a good surrogate for real seagrass, and that the presence of potential food (epiphytes) did not appear to influence habitat selection. There was also no difference in the habitat selected between day and night, and only minor differences with prawn size. Thus, juvenile M. latisulcatus appear to have a hierarchy of mechanisms for avoiding predators, with burying in sand being the preferred option. If burying is not possible, then seagrass is used for shelter. Active habitat selection to avoid predation appears likely to play a substantial role in determining the distribution of these animals on unvegetated sand- and mud-flats. PMID- 11399276 TI - Crepidula fornicata is not a first intermediate host for trematodes: who is? AB - Trematode larvae must generally invade a molluscan intermediate host, usually a gastropod, before they can reach reproductive maturity in another definitive host. The research literature to date has focused almost exclusively on the documented specificity between particular trematode species and particular molluscan hosts; little attention has been paid to gastropod species that do not appear to serve as hosts. We sampled Rhode Island and Massachusetts populations of the marine gastropod Crepidula fornicata to determine whether this widespread species serves as a first intermediate host for trematodes. We also sampled from the same habitat populations of Littorina littorea and Ilyanassa obsoleta, gastropods known to serve as first intermediate hosts for several trematode species. All individuals were examined by dissection for the presence of sporocysts, rediae, or developing cercariae. Although 4-28% of L. littorea (N=112) and I. obsoleta (N=84) were infected by larvae of at least one trematode species, no individuals of C. fornicata sampled from the same locations were so infected (N=136). A survey of the Biological Abstracts computer database indicates that snails in only about 10% of marine gastropod families are known to serve as first intermediate hosts for trematodes. We suggest that more attention be paid to marine gastropods that appear not to be infected by trematode miracidia. Such species may productively serve as new models for understanding trematode host specificity and gastropod resistance to infection. PMID- 11399277 TI - Isolation and identification of an ice-nucleating bacterium from the gills of the intertidal bivalve mollusc Geukensia demissa. AB - In the fall, freeze tolerant intertidal invertebrates usually produce ice nucleating proteins that are secreted into the hemolymph. These proteins help protect against freeze damage by insuring that ice formation is limited to extracellular spaces. Geukensia demissa, a freeze tolerant, salt marsh bivalve mollusc was examined for the presence of ice nucleating proteins. The ice nucleating temperature (INT) of the hemolymph was not significantly different from artificial seawater of the same salinity indicating the lack of an ice nucleating protein in the hemolymph. The palial fluid did have an elevated INT, indicating the presence of an ice nucleator. The INT of the palial fluid was significantly reduced by boiling and filtration through a 0.45-&mgr;m filter. High INT was also observed in the seawater associated with the bivalves, and was demonstrated in water samples collected from salt marshes but not sand and pebble beaches. Moreover, the INT of water samples collected from a salt marsh decreased in the summer. All of these data suggest that the ice-nucleating agents in the hemolymph and the seawater are ice-nucleating bacteria. One species of ice nucleating bacteria, Pseudomonas fulva was isolated from the gills of Geukensia. These bacteria could perform the same function as hemolymph ice-nucleating proteins by limiting ice formation to extracellular compartments. PMID- 11399278 TI - Effect of light on the feeding rates of pelagic and littoral mysid shrimps: a trade-off between feeding success and predation avoidance. AB - It has been shown that freshwater pelagic mysid shrimps capture zooplankton at a higher rate in light than in darkness. This has been suggested to be due to facilitation of visual predation on evasive zooplankton prey. To test this hypothesis with Baltic mysid shrimps, and to see whether pelagic (migrating) and littoral (non-migrating) mysids differ in this respect, we compared the feeding rates of Mysis mixta and Praunus flexuosus on the copepod Acartia spp. As light conditions change radically from the beginning of summer to late autumn at the Baltic latitudes, we conducted the experiments during three different times of the year to determine if there was a seasonal response to light in mysids. The feeding rates of pelagic mysids were significantly higher in total darkness than in light. In contrast, the feeding rates of littoral mysids did not differ in the dark and the light in the three different seasons. The decreased feeding of pelagic mysids under well-lit conditions may be an adaptation to avoid visual predation by pelagic fish. In contrast, littoral mysids, which live in the well lit layer throughout the day, do not show a similar response. The fact that light did not increase feeding in either of the mysid species indicates that these mysid species do not use vision in capturing prey, but rely mainly on mechano reception. PMID- 11399279 TI - Differences in the benthic-pelagic particle flux (biodeposition and sediment erosion) at intertidal sites with and without clam (Ruditapes philippinarum) cultivation in eastern China. AB - An annular flume or flux system (AFS) was deployed to measure the biodeposition and sediment resuspension processes at four stations in the mid-shore and low shore zones along both natural and Manila clam (Ruditapes philippinarum) farming transects in Xuejiadao intertidal area located in Jiaozhou Bay, eastern China. The results showed that there was a significant correlation between biodeposition rates and the density (r=0.984, P<0.05) and biomass (r=0.977, P<0.05) of the suspension feeding Manila clam. The effect of biodeposition at the farming transect was more intensive than at the natural transect where the biodeposition effect was lower than that resulting from natural sedimentation. In contrast, the biodeposition rate at the culture site with the high density of Manila clams was four times the sedimentation rate. Following the addition of Manila clams to increase their density in natural sediments, there was a marked increase in both clearance and biodeposition rates at all four sites. At the site with the highest natural bivalve biomass, the biodeposition rate increased 1.5-fold, while at the site with the least bivalve biomass, it increased by 40 times compared with the biodeposition rate by natural biota. The mean clearance rate of Manila clam was 0.90+/-0.34 l h(-1) ind.(-1) and the biodeposition rate was calculated to be 0.06+/-0.01 g h(-1) ind.(-1). The critical erosion velocities of intertidal sediment at Xuejiadao were from 17.4 to 20.4 cm s(-1). Relationships describing suspended particulate matter (SPM) vs. current velocity were analysed by linear regression following log transformation of the SPM. Statistical analysis of the slopes of the regression lines revealed that there were distinct differences between the low-shore and mid-shore (P<0.05) at both transects, respectively. There was also distinct difference between the two mid-shore stations (P<0.05) possibly due to differences in the densities of bioturbators (e.g. Macoma incongrua etc.), however, there was no significant difference between the two low shore stations (P>0.05). Furthermore, it was found that the site differences in sediment erodibility were not significantly correlated with measured physical properties of sediments and biota factors such as total macrofauna biomass, total abundance and macrofauna densities, Chl-a and Ph-a (P>0.05). However, there was a significant correlation between sediment erodability and both median grain size (P<0.01) and the density of the bioturbator M. incongrua (P<0.05). There was also evidence of a slight increase in sediment stability after 3 h of air exposure, but the effect was not overcome following the addition of Manila clams. It is hypothesised that the lower stability of sediments at the mid-shore level was probably associated with higher densities of bioturbators and with disturbance by shrimp farming near the high-shore. PMID- 11399280 TI - Hypertension and appraisal of physical and psychological stressors. AB - OBJECTIVE: In the operant conditioning of hypertension hypothesis, it is assumed that the frequently found diminished sensitivity to painful stimuli in hypertensives can be generalized to sensitivity to other stressors, including psychological stressors. The validity of this assumption is examined in the present study. METHODS: Unmedicated hypertensives (42) and normotensive controls (21) of both sexes were exposed to a physical stressor (electric current) and psychological active coping (mental arithmetic, free speech) and passive coping (unpleasant films) tasks, while indices of prestressor anxiety and task appraisal were measured. RESULTS: Hypertensive women, but not men, showed diminished pain sensitivity, together with lower prestressor anxiety and a tendency to lower negative appraisal of the tasks, compared to their normotensive counterparts. In addition, positive correlations were obtained between pain sensitivity and negative appraisal of psychological stressors involving interpersonal threat (speech) and passive coping (films). CONCLUSION: Preliminary support has been obtained for extrapolation of diminished pain appraisal to appraisal of some psychological stressors (although for a part only in women); an important assumption in the operant conditioning hypothesis of hypertension. PMID- 11399281 TI - Similar patterns of cardiovascular response during emotional activation as a function of affective valence and arousal and gender. AB - OBJECTIVE: Laboratory studies of emotion-induced cardiovascular responses have been conducted predominantly with a specific affects approach rather than a dimensional approach. The purpose of this study was to apply the principles of the Circumplex Model of Affect (i.e., valence and arousal) to investigate cardiovascular reactivity during emotional activation in men and women. METHODS: Forty-two healthy university students (mean age = 19.45, 52% women, 58% Caucasian) engaged in personally relevant recall tasks that varied as a function of valence and arousal. Self-reported valence and arousal, systolic and diastolic blood pressure (SBP and DBP, respectively), heart rate (HR), preejection period (PEP), stroke index (SI), cardiac index (CI), and total peripheral resistance (TPR) were measured during baseline and task periods. RESULTS: Cardiovascular responses were found to be largely comparable across the recall tasks and were characterized by significant increases in blood pressure, HR, and TPR, and decreases in SI (Ps < .001). In addition, SBP during negative valence tasks was significantly higher than during positive valence tasks (P < .03), and PEP lengthened more during low as compared to high arousal tasks (P < .03). CONCLUSIONS: These results highlight the similarity of hemodynamic adjustments during the verbal expression of emotion across gender and the dimensions of valence and arousal. The overall response pattern suggests alpha-adrenergically mediated sympathetic activation and vagal withdrawal. PMID- 11399282 TI - Factorial structure of the 20-item Toronto Alexithymia Scale: confirmatory factorial analyses in nonclinical and clinical samples. AB - OBJECTIVE: The 20-item Toronto Alexithymia Scale (TAS-20) measures three intercorrelated dimensions of alexithymia: (1) difficulties identifying feelings (DIF), (2) difficulties describing feelings (DDF), and (3) externally oriented thinking (EOT). The aim of the study was to test the three-factor model of the TAS-20 using confirmatory factorial analyses (CFA). METHOD: 769 healthy subjects and 659 patients meeting the DSM-IV criteria for substance use disorders or eating disorders completed the TAS-20. The correlation matrices for each of the samples were analyzed with LISREL 7.16. RESULTS: In each sample, the three-factor model was found to be replicable. CONCLUSION: The three TAS-20 subcales can be used to explore the distinct facets of the alexithymia construct. PMID- 11399283 TI - Relationship of gastric myoelectrical and cardiac parasympathetic activity to chemotherapy-induced nausea. AB - OBJECTIVES: We evaluated (a) whether pretreatment levels of gastric tachyarrhythmia, a dysrhythmic pattern of gastric myoelectrical activity, or cardiac parasympathetic activity are associated with the development of chemotherapy-induced nausea and (b) whether chemotherapy-induced nausea is preceded by an increase in gastric tachyarrhythmia and a decrease in cardiac parasympathetic activity, as has been observed during motion sickness. METHODS: Electrogastrograms and estimates of respiratory sinus arrhythmia (RSA) were obtained from cancer chemotherapy patients before treatment and for approximately 24 hours after treatment. RESULTS: Higher levels of pretreatment gastric tachyarrhythmia were observed on chemotherapy sessions that were followed by posttreatment reports of nausea. Pretreatment levels of RSA, however, did not differ between chemotherapy treatments that were and were not followed by nausea. No statistically significant changes in gastric tachyarrhythmia or RSA were observed prior to first reports of nausea following chemotherapy. CONCLUSIONS: In contrast to motion sickness, chemotherapy-induced nausea may not be related to an increase in dysrhythmic gastric myoelectrical activity; however, higher levels of pretreatment gastric tachyarrhythmia may be related to posttreatment reports of chemotherapy-induced nausea. PMID- 11399284 TI - Interactions of a history of migration with the course of pain disorder. AB - OBJECTIVE: Previous studies indicate that sociocultural factors affect the course of pain disorder. We investigated the role of nationality as an indicator of cultural factors, and of the degree of inclusion (DI) in Swiss society as an indicator of the migration process on the course of pain disorder. METHOD: In a semiprospective case-control study, outcome was assessed after a 2-year follow-up in 57 patients treated for pain disorder by phone interview with the patients. One Swiss patient was matched with two Spanish or Italian patients living in Switzerland. Patients' appraisal of course of illness (PACI) and of current general health (PACGH) were assessed as the sum score of their ratings of general well-being, handicap in everyday life, work ability, mood and pain. Family physicians were also asked to rate patient's course of illness. DI in Swiss society was assessed according to type of work permit, age at immigration and fluency in the language of the receiving country. RESULTS: Swiss patients evaluated their PACI [H(1,56) = 4.30; P = .0038] and PACGH [H(1,56) = 9.04; P = .003)] more favourably than patients from Italy and Spain. This difference was confirmed by the family physician's evaluation of the course of illness. A similar difference in outcome was found in favour of foreign patients with a higher DI. CONCLUSION: These results indicate that, in addition to sociocultural factors, the DI affects the outcome of pain disorder in patients with a history of migration. PMID- 11399285 TI - Gender differences in psychological distress among patients with irritable bowel syndrome. AB - OBJECTIVE: We examined possible gender differences in psychological distress in a sample of treatment-seeking Irritable Bowel Syndrome (IBS) patients. METHODS: A total of 341 IBS patients (238 females, 83 males) were studied. Structured psychiatric interviews were available on 250 participants. RESULTS: We found significantly higher scores for females than males on the Beck Depression Inventory (BDI), Trait Anxiety of the State-Trait Anxiety Inventory (STAI), and Scales 2 and 3 of the MMPI. However, there were no differences in the percentages of the two samples meeting criteria for one or more Axis I psychiatric disorders, with 65.6% of the total sample meeting these criteria. CONCLUSION: Gender differences in psychological distress appear to be a function of method of measurement. PMID- 11399286 TI - Associations between coping and survival time of adult leukemia patients receiving allogeneic bone marrow transplantation: results of a prospective study. AB - BACKGROUND: To investigate associations between coping strategies and length of survival in a sample of 52 adult leukemia patients receiving allogeneic bone marrow transplantation (BMT). METHODS: 52 adult patients, diagnosed with acute (AML) and chronic myeloid leukemia (CML) admitted for allogeneic BMT to a university hospital BMT unit in preparation for a transplantation of genotypically matched HLA donor marrow, were interviewed immediately after informed consent and prior to preparatory treatment for transplantation. Semistructured interviews were conducted and recorded for analysis to assess coping styles and were evaluated by a new content analytic coping measure [Ulm Coping Manual (UCM)]. Patients were a random sample of all eligible patients on the BMT unit between May 1990 and May 1994. RESULTS: Complete audiotaped interviews were rated by blind raters, employing a newly developed content analysis for the identification of patients' coping strategies. Multivariate analysis using a Cox model revealed three pretransplant variables that demonstrated a statistically significant influence on 5-year survival: Stage of Disease at transplant (P < .012), Distraction (P < .007), and Fighting Spirit as coping modalities (P < .013). CONCLUSIONS: The results of this prospective study document the impact of certain psychological variables, notably coping style on survival with BMT. This suggests the necessity of utilizing psychosocial interventions to address stress and anxiety in patients awaiting transplantation in order to reduce anxieties and to employ more effective coping techniques to deal more appropriately with their situation and to enhance Fighting Spirit. The effects on survival of such psychosocial interventions need to be tested in a randomized controlled study. PMID- 11399287 TI - Mind matters. Coping and cancer progression. AB - The idea that having an 'attitude' about cancer makes a difference in its course is a popular but controversial one. Most oncologists and surgeons believe that tumor type and stage, general health, and medical treatment are all that account for the variance in outcome. Many patients and their families believe that having the right attitude makes a difference in the course of disease. This leads us to two empirical questions: (1) Does coping make a difference in disease progression when medical prognostic variables are taken into account? and (2) What constitutes the 'right attitude'? PMID- 11399288 TI - Effect of water supply method and flow rate on drinking behavior and fluid balance in horses. AB - This study investigated three methods of water supply on drinking preference and behavior in six Standardbred geldings (2-9 years, 505+/-9 kg). The water sources were buckets (B), pressure valve (PV), and float valve (FV) bowls. In an initial drinking preference test, PV was tested at three flow rates: 3, 8, and 16 l/min (PV3, PV8, and PV16), and FV at 3 l/min (FV3). Water intake was measured in l and presented as the percentage of the total daily water intake from each of two simultaneously presented alternatives. The intake from PV8 was greater than from both PV3 (72+/-11% vs. 28+/-11%) and PV16 (90+/-4% vs. 10+/-4%). All horses showed a strong preference for B, 98+/-1% of the intake compared to 2+/-1% from PV8. Individual variation in the data gave no significant difference in preference between the two automatic bowls. In the second part of the study, drinking behavior and fluid balance were investigated when the horses drank from FV3, PV8, and B for 7 consecutive days in a changeover design. Despite a tendency for an increase in total daily drinking time from FV3, the daily water intake was significantly lower (43+/-3 ml/kg) than from PV8 (54+/-2 ml/kg) and B (58+/-3 ml/kg). Daily net water gain [intake-(fecal+urinary output)] was only 0.5+/-3 ml/kg with FV3, resulting in a negative fluid balance if insensible losses are included. These results show that the water supply method can affect both drinking behavior and fluid balance in the horse. PMID- 11399289 TI - A nonextinction procedure for long-term studies of classically conditioned enhancement of acoustic startle in the rat. AB - In the Fear-Potentiated Startle (FPS) paradigm, the reflexive response to a noise burst is enhanced when it is presented with a stimulus (typically a light) that previously had been paired with the presentation of an aversive stimulus, usually an electric shock. In the FPS paradigm, the conditioned effect is demonstrated under conditions of extinction, i.e., the light is not paired with the shock during FPS testing. Because of this, the FPS paradigm is of somewhat limited value as a longitudinal measure for studying classically conditioned enhancement of acoustic startle. The present studies report a simple and reliable nonshock procedure for studying classically conditioned potentiation of acoustic startle in the rat that does not utilize testing under conditions of extinction. Naive rats were exposed to 5-or 3-days/week startle test sessions for up to 20 weeks. Twenty (20) startle stimuli (115 dB noise bursts; 40 ms in duration) were presented during each session. Half of these startle stimuli were presented in darkness and half were immediately preceded by a 3500-ms presentation of a 15-W light. With this paradigm, the effects of pairing the light with the startle noise burst could be studied across many test sessions in the absence of extinction training. The light did not increase startle amplitude on the first few startle trials of the first test session. By the end of the first session, however, and continuing for many weeks of testing, startle responses in the presence of the light were significantly greater (by 30-40%) than in the absence of the light. The finding that the startle stimulus itself can serve as an unconditioned stimulus (UCS) to enhance subsequent startle responses replicates an earlier finding. The persistence of this Startle-Potentiated Startle (SPS) effect across multiple weeks of testing is in contrast to that earlier report. PMID- 11399290 TI - Protection against radiation-induced conditioned taste aversion by Centella asiatica. AB - Radiations are known to cause behavioural perturbations like conditioned taste aversion (CTA), performance decrement, learning, etc., even at very low doses. The manifestation of radiation-induced behavioural degradation has not been understood well and requires further studies. Therefore, the effects of low-dose whole-body 60Co gamma-irradiation in male rats were studied in terms of body weight and CTA learning. For CTA, the consumption of saccharin solution was considered as a parameter. To protect against the adverse effects of radiation, Centella asiatica (aqueous extract) was tested and compared with ondansetron, a standard antiemetic drug. A dose of 2 Gy incurred significant body weight loss [t(9)=9.00, P<.05] and induced CTA in rats [t(26)=9.344, P<.01]. Administration of C. asiatica (100 mg/kg bw ip, 2 Gy, -1 h) rendered significant radioprotection against radiation-induced body weight loss and CTA that became evident on the second postirradiation day [t(7)=0.917, P>>.05; t(7)=4.016, P>.05]. Ondansetron (1 mg/kg bw) elicited higher degree of protection against CTA [t(7)=3.641, P>.05] than C. asiatica [t(7)=7.196, P>.05] on the first postirradiation day, but on the second postirradiation day, both were equally effective [t(7)=3.38, P>.05; t(7)=4.01, P>.05]. In case of C. asiatica-treated animals, however, there was a consistently declining CTA from the second to the fifth postirradiation day whereas in ondansetron-treated animals it was inconsistent. Present investigation suggests that C. asiatica could be useful in preventing radiation-induced behavioural changes during clinical radiotherapy. PMID- 11399291 TI - Oral irritant properties of menthol: sensitizing and desensitizing effects of repeated application and cross-desensitization to nicotine. AB - The irritant properties of menthol and its interactions with nicotine were investigated psychophysically in human subjects. In the first experiment, 0.3% L menthol was applied successively to one side of the tongue 10 times at a 1-min interval (30-s interstimulus interval, ISI), and subjects rated the intensity of the perceived irritation. The intensity of irritation progressively decreased across trials, consistent with desensitization. To test for cross-desensitization of nicotine-evoked irritation by menthol, nicotine (0.6%) was applied to both sides of the tongue simultaneously, 5 min after the conclusion of menthol application. Using both a two-alternative forced choice (2-AFC) paradigm, and also by obtaining independent ratings of the irritant intensity on each side of the tongue, it was found that nicotine-evoked irritation was significantly weaker on the menthol-pretreated side. To control for a possible confounding effect of cooling, nicotine was applied bilaterally only after the cooling sensation of menthol had subsided. Nicotine-induced irritation was still significantly weaker on the menthol-pretreated side, consistent with cross-desensitization of nicotine evoked irritation by menthol. In a final experiment, menthol was repeatedly applied to one side of the tongue at a shorter (20 s) interval (5-s ISI), and elicited a rapid increase in irritant sensation over the initial trials, consistent with sensitization, followed in subsequent trials by a progressive reduction in irritation (desensitization). After a 5-min rest period, self desensitization was confirmed. Repeated application of menthol at the same short ISI was then resumed, and resulted in a significant mean increase in irritant intensity consistent with stimulus-induced recovery (SIR). PMID- 11399292 TI - Task-dependent strain difference of spatial learning in C57BL/6N and BALB/c mice. AB - In the present study, we used a dry maze task to assess the spatial learning ability of C57BL/6N and BALB/cA mice besides the water maze task. In Experiment 1, the performance of C57BL/6N and BALB/cA mice in the water maze task and dry maze task were investigated. In the former task, the mice had to learn the position of a hidden platform submerged below the water surface and they had to learn the position of a baited hole on the circular maze in the latter task. C57BL/6N mice showed significant learning in both maze tasks, whereas BALB/cA mice showed learning in a dry maze but not in water maze as reported before. In Experiment 2, a dry maze task was conducted on a circular open field which contained 16 holes arranged symmetrically and which was put on the same height as the surface of the water maze. In Experiment 2, BALB/cA mice showed significant improvement in latency and path length to reach the food hole. The poor performance of the BALB/cA in the water maze task may reflect a weak motivation, escaping from water, rather than the poor spatial memory in this strain. PMID- 11399293 TI - Subchronic mild noise stress increases HRP permeability in rat small intestine in vitro. AB - Recently we reported an increased trans- and paracellular protein permeability in rat small intestine after acute cold restraint stress. In the present study, we applied randomized 95- or 105-dB white noise pulses during 45 min/h, 12 h/day, duration 8 days, as a milder, but more chronic stressor to male rats. At 8 days before the noise experiments, 50% of the animals were cannulated in the vena cava for blood sampling during the experimental period. The other 50% of the animals were sacrificed at Day 9, segments of ileum were mounted in Ussing chambers and perfused at 37 degrees C. Horseradish peroxidase (HRP) was added mucosally, serosal appearance was detected enzymatically and tissues were fixed for electron microscopy. In the animals exposed to 95-dB noise, plasma corticosterone levels were enhanced twofold compared to controls, and ileal HRP flux was enhanced twofold. Electron micrographs of tissue from stressed or control animals showed no detectable paracellular staining of HRP. Quantification of HRP-containing endosomes in enterocytes revealed a twofold increase in endosome number in the animals exposed to 95-db noise indicating that the increased HRP permeability was primarily due to increased endocytosis. In contrast to the animals exposed to 95 dB noise, rats exposed to 105-dB noise showed no increase in corticosterone levels and ileal HRP fluxes were not significantly different from controls. We conclude that mild subchronic noise stress may cause a decrease in intestinal barrier function by increased transcytosis of luminal antigens. PMID- 11399294 TI - Interactions of inflammatory pain and morphine in infant rats: long-term behavioral effects. AB - Neonatal rat pups exposed to repetitive acute pain show decreases in pain threshold and altered behavior during adulthood. A model using prolonged inflammatory pain in neonatal rats may have greater clinical relevance for investigating the long-term behavioral effects of neonatal pain in ex-preterm neonates. Neonatal rat pups were exposed to repeated formalin injections on postnatal (P) days 1-7 (P1-P7), with or without morphine pretreatment, and were compared with untreated controls. Behavioral testing during adulthood assessed pain thresholds using hot-plate (HP) and tail-flick (TF) tests, alcohol preference, and locomotor activity (baseline and postamphetamine). Adult rats exposed to neonatal inflammatory pain exhibited longer HP latencies than controls and male rats had longer HP thresholds compared to females. Male rats exposed to neonatal morphine alone exhibited longer TF latencies than controls. Both neonatal morphine treatment and neonatal inflammatory pain decreased ethanol preference, but their effects were not additive. During adulthood, male rats exposed to neonatal inflammatory pain exhibited less locomotor activity than untreated controls. We conclude that neonatal formalin and morphine treatment have specific patterns of long-term behavioral effects in adulthood, some of which are attenuated when the two treatments are combined. PMID- 11399295 TI - Intrathecal methiothepin and thyrotropin-releasing hormone effects on penile erection. AB - Intrathecal thyrotropin-releasing hormone (TRH) potently inhibits penile erection at all doses (100, 500, 1000 or 5000 pmol) tested so far. Since the serotonin receptor antagonist methiothepin (MT) inhibits TRH responses in other systems, this study tested the hypothesis that MT-sensitive receptors mediate the effect of TRH on penile erection in rats. When compared to controls, the highest doses of IT TRH (0, 10 or 500 pmol) or MT (5 or 50 nmol) significantly altered penile reflex latency. When coadministered (50 nmol MT/500 pmol TRH), the effect of TRH was reversed, suggesting that the high dose of MT antagonized the inhibitory actions of TRH. The low dose of MT (5 nmol) did not block the 500 pmol TRH inhibition of reflex latency. These data further suggest that MT sensitive receptors are important in (1) mediating normal penile reflexes and (2) mediating the inhibitory response to TRH. PMID- 11399296 TI - Feeding, drinking, and temperature responses of chickens to intracerebroventricular histamine. AB - The present study examines the effects of intracerebroventricular injections of histamine (HA) and two HA antagonists, the H(1) receptor antagonist chloropheneramine maleate (CM) and the H(2) receptor antagonist cimetidine (CIM), on food and water consumption and body temperature in chickens. Single-Comb White Leghorns (SCWL) and broiler cockerels were utilized for these experiments. The first pair of experiments consisted of intracerebroventricular injections of HA and its effects on food and water consumption. HA was infused at dosages of 0, 25, 50, and 100 microg/10 microl of artificial cerebrospinal fluid (aCSF). HA significantly decreased food and water intake in a dose-dependent manner. The second pair of experiments examined the effects of HA on water intake while birds had no access to feed. Water intake was not significantly affected by intracerebroventricular injections of HA. The next pair of experiments examined the effects of HA on body temperature. In SCWL, body temperature was not affected by HA until 165 min postinjection when HA decreased temperature in a quadratic dose-response with maximum hypothermia being achieved at a dose of 25 microg. In contrast, HA increased body temperature in broilers beginning at 75 min postinjection. In the final series of experiments, the anorexia induced by HA was attenuated in SCWL and broilers with pretreatment of either CM or CIM. These results suggest that HA has an anorexigenic effect in SCWL and broiler cockerels, and this effect is mediated by both H(1) and H(2) receptors. Water intake is not directly affected by the intracerebroventricular injection of HA. Whereas HA increased body temperature in broilers, the response in SCWL is equivocal. PMID- 11399297 TI - Fos-like immunoreactivity in brain regions of domestic rams following exposure to rams or ewes. AB - Limbic and basal forebrain-hypothalamic regions from male sheep differing in sexual performance were quantified for fos-like immunoreactivity. Rams classified as high-sexually performing (HP), low-sexually performing (LP), and male-oriented (MO) received noncontact sensory stimulation from either ewes in estrus (HP, n=5; LP, n=4; MO, n=4) or other males (HP, n=5; LP, n=4; MO, n=5) for a 4-h period on each of 3 consecutive days. Following exposure to stimulus animals on the third day, rams were euthanized and their brains were perfused with a 1% paraformaldehyde/1.5% glutaraldehyde solution and sections were analyzed for fos like immunoreactivity. Brain regions analyzed were the medial amygdala (meAMY), medial preoptic area (mPOA), bed nucleus of the stria terminalis (BNST), and ventromedial hypothalamic nucleus (VMH). Fos-like immunoreactivity differed between groups in the mPOA and BNST but not in the meAMY or VMH. LP rams exposed to estrous ewes had more (P<.05) neurons staining positive for fos and fos related antigens (FRA) in the mPOA and BNST than LP rams exposed to other rams or MO rams exposed to either estrous ewes or other rams. Numbers of neurons staining positive for FRA in the mPOA and BNST of LP rams exposed to estrous ewes, however, were not different (P>.05) from HP rams exposed to either estrous ewes or other rams. The similar fos-like immunoreactivity in areas important for the display of sexual behavior in HP and LP rams may reflect similar sensory input in these two groups of rams; however, LP rams, in contrast to HP rams, do not appear to respond similarly to the same sensory stimulus. PMID- 11399298 TI - Long-term sensitization of cardiovascular stress responses after a single stressful experience. AB - There is evidence that the experience of traumatic events may play a role in the pathogenesis of somatic diseases, including cardiovascular disorders. In this study, telemetry was used to investigate the long-term effects of a single stressful experience on cardiovascular and behavioral responses to novel challenges 2 weeks later. Rats were exposed to footshocks and tested for sensitization using the following challenges: novel cylinder (Day 14); shock prod acquisition test (Day 15); and shock prod retention test (Day 16). No difference in basal somatomotor activity (SA), heart rate (HR) and blood pressure between preshocked rats and control rats was found. However, preshocked rats displayed an enhanced blood pressure response compared to controls during the shock prod acquisition test and the shock prod retention test. No differential increase in HR response between both groups was found. During the novel cylinder test, the preshocked rats displayed less SA while no behavioral differences were found in the shock prod acquisition test and the shock prod retention test. We conclude that a single stressful experience induces long-term sensitization of blood pressure responses to novel challenges that are not necessarily linked to sensitized behavioral responses. The footshock model may be a useful model to study autonomic hyperresponsivity found in posttraumatic stress disorder (PTSD). PMID- 11399299 TI - Effects of arteether on an auditory radial-arm maze task in rats. AB - We evaluated the behavioral and neural toxicity of the artemisinin antimalarial compound, arteether (AE), using a novel radial-arm maze procedure. We have previously shown that AE can produce a distinctive pattern of neurotoxicity in the brainstem and that auditory nuclei are particularly vulnerable. Thus, we assessed performance which depended upon auditory processing. We trained rats to choose one of eight arms of a radial maze, depending upon which arm served as the source of a white noise stimulus. Correct responses produced food reinforcement while incorrect choices had no programmed consequences. When the task was acquired, AE (25 mg/kg/day; n=7) or oil vehicle (n=7) was administered (intramuscularly) for seven consecutive days. Behavioral sessions were conducted during the days of drug administrations and for 7 days following drug administrations. Subsequently, histopathology was conducted and a quantitative assessment of the nucleus trapezoideus was made. AE produced a progressive deficit in performance on the maze task. That is, accuracy decreased, choice latency increased, and the number of trials completed decreased. Moreover, the greatest deficits were observed during the period following drug administrations. AE-treated rats revealed marked damage in the nucleus trapezoideus. The damage included chromatolysis, necrosis, and gliosis. Vehicle-treated rats did not show performance deficits or neuropathology. These results extend earlier studies and show that AE can produce damage in the n. trapezoideus of rats, which is associated with performance deficits on a complex auditory task. Thus, the auditory radial-arm maze task is a useful tool for assessing AE-induced toxicity. PMID- 11399300 TI - Prenatal morphine exposure differentially alters learning and memory in male and female rats. AB - The present study tested the hypothesis that exposure to morphine on prenatal days 11-18 impairs performance on tasks requiring learning and memory in adult male and female rats. In Experiment 1, a symmetrical maze was used to measure learning. In Experiment 2, an eight-arm radial maze was used to assess working spatial memory. The results of Experiment 1 demonstrated that prenatal morphine exposure reduces the time needed to complete the trials, but does not affect the accuracy of performance in male rats. In contrast, prenatal drug treatment had no effects on either the time or the accuracy of performance in female rats. In Experiment 2, both male and female morphine-exposed rats needed more time to complete regular trials (no delay) than controls; however, morphine exposure in male rats did not affect performance on tasks requiring memory, measured with delay trials, but hindered it in ovariohysterectomized (OVX) female rats. In OVX females, replacement injections of both estrogen and progesterone restored the impairment of performance on delay trials produced by prenatal morphine exposure. Thus, the present study demonstrates that prenatal morphine exposure differentially alters performance of adult male and female rats on tasks requiring learning and spatial memory. PMID- 11399301 TI - Effect of short-term fasting on macronutrient self-selection in sea bass. AB - The influence of two different fasting periods on macronutrient self-selection during refeeding was studied in sea bass. Twelve groups of three sea bass were kept under laboratory conditions with a 12:12 light-dark cycle and at 22.5 degrees C. Each aquarium was provided with three self-feeders containing three different diets made up of two macronutrients: protein-fat, protein-carbohydrate, and carbohydrate-fat. The fish were allowed to acclimatise to the diets and laboratory conditions in 4 weeks, during which they selected a baseline diet of 51% protein, 16.5% carbohydrate, and 32.5% fat, in terms of digestible energy. The fish were then deprived of food for 6 days, after which they were permitted to refeed. When food demands stabilised, they were subjected to total food deprivation for another 2 weeks. The total energy demanded from all the macronutrients increased significantly during the first day after both fasting periods. The proportion of macronutrient self-selection after fasting periods remained unchanged in respect to the baseline, except for protein energy which remained high in the two first days of refeeding after the second fasting period (the 51% of total demanded energy from protein during the baseline period rising to 60%). In short, after two short fasting periods of different length, sea bass showed a compensatory ingestion of energy, with a slightly increased demand from protein after the longest fasting period. PMID- 11399302 TI - A comparison of food preferences and nutrient composition in captive squirrel monkeys, Saimiri sciureus, and pigtail macaques, Macaca nemestrina. AB - I assessed the occurrence of food preferences in captive squirrel monkeys and pigtail macaques and analyzed whether their preferences correlate with nutrient composition. Using a two-alternative choice test, I repeatedly presented six Saimiri sciureus and six Macaca nemestrina with all possible binary combinations of 12 types of food that are part of their diet in captivity. The two species exhibited significantly different rank orders of preference. Correlational analyses revealed that the preference ranking of the squirrel monkeys was significantly positively correlated with total energy content, irrespective of the source of energy as neither total carbohydrate content nor protein or lipid content was significantly correlated with food preference. In contrast, the preference ranking of the pigtail macaques showed a significant positive correlation with total carbohydrate content and with fructose content but not with total energy content of the food items. These results suggest that squirrel monkeys are opportunistic feeders with regard to maximizing net gain of energy, whereas pigtail macaques are not but rather seek to meet their requirements of metabolic energy by preferring foods that are high in carbohydrates. PMID- 11399303 TI - Female meadow voles housed in long and short daylengths respond to exogenous estrogen with similar mating latencies. AB - After weaning, adult female meadow voles were maintained for 7 weeks in either long (LD, 14 h light/day) or short photoperiods (SD, 10 h light/day). They were then ovariectomized and implanted with 3-week, timed-release estrogen pellets (0.0, 0.001, 0.05, or 0.5 mg/pellet of 17-beta-estradiol). An additional group received a sham ovariectomy (intact) and a 0.0-mg/pellet control (no estrogen) pellet. One week after surgery, females were paired with an LD sexually experienced male. Each pair was videotaped continuously until the first intromission or for 2 weeks. LD sham animals mated significantly earlier than did SD sham animals (P=.05). However, there were no differences in mating latencies between LD and SD control groups or between any of the LD and SD groups receiving estrogen replacement (P>.05). In addition, no ovariectomized animals receiving either the control or the 0.001-mg/pellet estrogen dosage mated within the 2-week time period, while all shams and all animals receiving either the 0.05- or 0.5 mg/pellet doses mated. The results indicate that there is some minimal amount of estrogen that is necessary for meadow voles to enter behavioral estrus and that LD and SD females do not differ in their sensitivity to estrogen, since animals in both photoperiods mate with similar latencies as long as enough estrogen is present. PMID- 11399304 TI - Modulating effects of prenatal stress on hyperthermia induced in adult rat offspring by restraint or LPS-induced stress. AB - The present study was carried out to investigate the effects of prenatal stress on stress-induced hyperthermia in adult rats. Prenatal stress was administered daily for 3 days (embryonic days 15-17) by restraining pregnant rats in a small cage either for 30 or 240 min. After birth, foster mothers raised the pups. Offspring were tested at 9-10-weeks-old. Changes in body temperature and in the plasma concentrations of corticosterone, norepinephrine (NE), and epinephrine (Epi) induced by restraint or lipopolysaccharide (LPS)-induced stress were examined. By comparison with the prenatally nonstressed control group, the 240 min stress group showed a significantly lower hyperthermia in response to restraint stress but a higher fever after injection of LPS. The 30-min stress group showed similar alterations in these hyperthermic responses but did not reach significance. Both the restraint stress and the injection of LPS evoked greater increases in the plasma level of corticosterone in the 240-min stress group than in the control group. Although restraint stress induced significant increases in NE and Epi in the control and 30-min stress groups, the plasma levels of these catecholamines did not increase in the 240-min stress group. These results demonstrate for the first time that prenatal stress has opposite effects on the hyperthermic responses to restraint and LPS injection, suggesting that different mechanisms underlie the modulating effects of prenatal stress on the responses to the two types of stressors. PMID- 11399305 TI - Impact of transgenic procedures on behavioral and physiological responses in postweaning mice. AB - This study evaluates the effects of biotechnological procedures involved in the process of microinjection-induced transgenesis in the mouse by comparing four groups of C57BL/6 mice that differ in their transgenic background (transgenics after integration of a functional corticotropin-releasing factor (CRF) gene construct, transgenics after integration of a nonfunctional CRF gene construct, nontransgenics after transgenic procedures, and controls). These four groups have been tested in various behavioral paradigms. Moreover, the development in growth, morphological characteristics, and clinical appearance of the animals have been recorded from 4 till 30 weeks of age. Differences in behavior, weight gain, and morphology were found between Group 1 (transgenic CRF animals) and Group 4 (control animals). For Group 2 (animals with a noncoding construct) and Group 3 (nontransgenic animals after transgenic procedures), no significant differences from control animals were found. This indicates that, under the present conditions, the biotechnological procedures related to transgenesis (microinjection, in vitro culture, and embryo transfer) have no significant effect on the normal development of the mice in the postweaning period. These results substantiate previous findings on these animals, obtained by screening them in the preweaning period (Days 0-21). However, before general conclusions as to what extent the technique of transgenesis affects the welfare of the animals can be drawn, more and different transgenic lines should be studied in this or a similar way. PMID- 11399306 TI - Long-term effects of social stress on antiviral immunity in pigs. AB - Mixing of unfamiliar pigs is common practice in intensive pig husbandry. Since pigs maintain a dominance hierarchy, mixing often leads to vigorous fighting. Apart from the negative impact that fighting has on welfare, there is evidence that the social stress associated with fighting suppresses immune function. In the present experiment, we investigated the impact of mixing on specific long term immune responses and protection against challenge infection after vaccination with pseudorabies virus (PRV). Specific pathogen-free (SPF) pigs were mixed pairwise with an unfamiliar same-gender conspecific or left undisturbed with a same-gender littermate at 3 days after vaccination with PRV. Half of the pigs were females (gilts) and half were castrated males (barrows). Mixing increased agonistic behavior to the same degree in gilts and barrows. Cortisol concentrations in saliva and catecholamine excretion in urine were increased in mixed pigs, and these effects were independent of dominance status and gender. Subsequently, the effects of mixing, gender, dominance status and interactions between these factors on immune response parameters were studied. The main result was that mixed barrows showed suppressed immune responses after vaccination and increased clinical symptoms after challenge infection compared to control barrows. Mixed gilts however did not differ from control gilts. It also appeared that mixed dominants were more seriously affected than mixed subordinates were. We conclude that, in some pigs, social stress after mixing suppresses the immune response to a viral vaccine and consequently impairs protection against challenge infection. PMID- 11399307 TI - Development and consistency of gait in the mouse. AB - Mouse models of human disease may display developmental abnormalities or adult onset of the condition. Since many diseases are accompanied by gait disturbances, knowledge of normal gait development in the mouse and its adult characteristics might be valuable as standards against which to appraise disease progression and the efficacy of putative therapies. Assessment of the gait of mice from postnatal day (pnd) 13 to postnatal week (pnw) 80 was undertaken utilising video techniques to examine velocity, stride, stance and swing times and between pnw 29 and 80 using load cells for analysis of the vertical reaction force (P(z)) associated with limb placements. Some adult features are apparent by pnd 13, but in the hindlimb (HL) particularly, the adult pattern of relationships between stride, stance and swing are not established. Adult characteristics of forelimb (FL) deployment develop earlier than those of HL while the systems controlling HL stance develop earlier than those regulating its swing. All the features measured, however, such as the shorter stance and longer swing of FL compared to HL, are established in their adult form by pnd 24 and maintained throughout adult life. In healthy mice at pnw 80, there was no deviation from the adult pattern of gait in which P(z) transmitted via FL exceeds that via HL by around 5%. We did not detect any significant change in any other variable or in their relationships. PMID- 11399308 TI - Social stress paradigms in male mice: Variations in behavior, stress and immunology. AB - Male OF1 strain mice were allocated, after 2 weeks of individual housing, to cohabitating (6 or 16 days), fixed dyadic interaction pairs (6 or 16 daily encounters) or control groups (6 or 16 days). These different social stress situations were assessed for their effects on splenic contents of NE, IL-1 and IL 2 and serum levels of corticosterone. Spleen NE contents showed no significant variations, but serum corticosterone titers were generally higher in interacting pairs and subordinates. Splenic IL-2 did not respond in the same way to the treatments as IL-1. The differences in splenic interleukin contents could not be simply related to observed changes in serum corticosterone levels. Different mechanisms appear to regulate changes in glucocorticoids and the measured cytokines. These physiological phenomena do not simply reflect in the animal's social status (dominant or submissive). The intensity and duration of the agonistic behavior displayed as well as the interaction experience accumulated may account for the observed differences between the paradigms. PMID- 11399309 TI - Circadian phase shifting: Relationships between photic and nonphotic phase response curves. AB - A variety of photic and nonphotic stimuli can phase-shift the mammalian circadian pacemaker. It has been suggested that the phase-response curves (PRCs) characterizing these diverse stimuli may comprise two major PRC families, one typified by the photic PRC describing the response to brief light pulses, and the other typified by the nonphotic PRC describing the response to stimuli evoking behavioral arousal and/or locomotor activity. Additionally, the mammalian circadian pacemaker can be phase-shifted by dark pulses presented on a constant light (LL) background. While dark pulse-induced phase shifting was interpreted originally as a mirror-image photic effect, other observations suggest that the dark pulse PRC may instead belong to the family of nonphotic, activity-dependent PRCs. In a recent study, we reexamined the phase-shifting effects of dark pulses in the Syrian hamster, and concluded that the dark pulse PRC reflects both nonphotic and photic mirror-image mechanisms. In the current report, we reanalyze previously published hamster PRC data using polynomial curve-fitting procedures. The results of these analyses reveal that (a) the photic and nonphotic PRCs have identical shape but opposite phasing, and (b) the dark pulse PRC can be modeled by simple summation of nonphotic and photic mirror-image PRCs. This model predicts accurately the shape of the dark pulse PRC, particularly the extension of the phase-advance region into the subjective night. PMID- 11399310 TI - Genetic deletions of NCAM and PSA impair circadian function in the mouse. AB - The adult suprachiasmatic nuclei (SCN) express neural cell adhesion molecule (NCAM) that carries polysialic acid (PSA), a carbohydrate polymer which controls plastic cell-cell interactions in neural tissues. Expression of the three major isoforms of NCAM [180 (principal PSA carrier), 140, and 120 kDa) varies with developmental and physiological state. The requirements for NCAM and PSA in circadian timekeeping function were assessed among three transgenic mutant strains with differing expressions of PSA and NCAM. These included: NCAM(tm3Ciw) mice lacking only NCAM 180; NCAM(tm1Cwr) mice lacking NCAM 180, 140, and PSA; and NCAM(tm1Cgn) mice lacking all NCAM isoforms and PSA. Locomotor activity was stably entrained to a 12-h light/dark cycle (LD) in the NCAM(tm3Ciw) and NCAM(tm1Cwr) mutants, but not in NCAM(tm1Cgn) mutants, where the daily onset and offset of activity were irregular or lost. Under constant darkness (DD), the free running rhythmicity of the NCAM(tm3Ciw) mutants expressing PSA was similar to wild-type controls, but was markedly disrupted in the majority of the NCAM(tm1Cwr) and NCAM(tm1Cgn) mutants lacking PSA. This analysis indicates that PSA and its NCAM carrier are necessary for stable free-running circadian rhythmicity under DD, and that NCAM, but not PSA, is needed for synchronization to LD. Notably, when NCAM 180 is deleted, NCAM 140 compensates as the PSA carrier, and may assume other SCN-related functions of the NCAM 180 isoform. PMID- 11399311 TI - Determination of auditory thresholds in the brushtail possum (Trichosurus vulpecula). AB - The auditory abilities of the brushtail possum (Trichosurus vulpecula) have been measured by cochlear potential readings, but no behavioural determinations of their auditory abilities have been published. Six experimentally naive possums were trained to perform a two-response conditional discrimination between the presence and absence of an 880-Hz tone (at 80 dB). All six possums readily learned this task. The behavioural threshold for this tone was determined using a modified tracking procedure and found to be similar to that reported using cochlear microphonic potentials. One concern with the current method was the communal nature of the experimental environment so a further threshold determination in a sound-attenuating chamber was conducted. No substantial difference was noted between the results obtained in the two threshold determinations. The success of the current method means that a full, behavioural audiogram for the brushtail possum, which would complement the existing cochlear potential data, is now possible. PMID- 11399312 TI - Performance more than working memory disrupted by acute systemic inflammation in rats in appetitive tasks. AB - Evidence from molecular biology, epidemiology, behavioral pharmacology, and clinical science support the conclusion that brain inflammation contributes to the pathogenesis of cognitive symptoms in Alzheimer's disease (AD) and other neuropsychological disorders. Three different tests were conducted to determine whether the acute inflammatory response induced by systemic lipopolysaccharide (LPS) treatment is accompanied by a selective disruption of working memory functioning in rats. Doses of LPS sufficient to induce a thermoregulatory response were administered intraperitoneally and their effects on behavioral measures of symbolic working memory, spatial learning, and spatial memory consolidation, were assessed. LPS-induced immune activation was found not to significantly affect memory processes in any of the behavioral tests used. However, LPS-induced immune activation caused performance deficits consistent with a disruptive effect of LPS on motivation and arousal. These results suggest that sickness behavior induced by immune stimulation is not necessarily accompanied by selective impairment in memory processes. The importance of distinguishing cognitive disruption from performance impairment in interpreting the behavioral effects of inflammatory mediators is discussed. PMID- 11399313 TI - The role of cholecystokinin in the expression of seasonal variation in the feed intake and eating pattern of red deer (Cervus elaphus). AB - The role of cholecystokinin (CCK) in the expression of seasonal variation in feed intake and feeding activity in ruminants was investigated by active immunisation of castrated male red deer (Cervus elaphus). In April, animals of two groups (five animals per group) were immunised against either CCK or vehicle solution only and booster injections were administered at 2-month intervals for the following year. Measurements were conducted for a period of more than a year from July. There were no significant effects of immunisation on mean daily food intake. However, there was a significant interaction (P<.01) between immunisation and month (season), with respect to rate of feed ingestion during meals, with animals immunised against CCK exhibiting higher mean rates of ingestion during October to May but lower mean rates during June to September. It is concluded that systemic CCK has a role in the expression of seasonal variation in the rate of feed ingestion during meals in ruminants and that this, in turn, may affect the pattern of seasonal change in daily feed intake. Since there was no evidence of differences with treatment in profiles of insulin, growth hormone (GH), thyroxine (T(4)), triiodothyronine (T(3)), insulin-like growth factor-1 (IGF-1) or prolactin, it is unlikely that this effect is expressed through changes in the profiles of these hormones. PMID- 11399314 TI - Endogenous plasma testosterone levels and commission errors in women: A preliminary report. AB - A correlation between elevated testosterone and aggressive behavior has been demonstrated in animals and to a lesser degree in humans, primarily in the context of dominance. Some aggression, namely non-premeditated aggression, is characterized by impaired impulse control. Real-world aggressive histories and self-reported impulsivity have correlated with commission errors (failures to withhold responses to nontarget stimuli) in versions of the continuous performance test (CPT). To begin exploring whether testosterone may play a role in aggression due more to a direct relationship with impaired impulse control, we related plasma total testosterone concentrations of 27 psychiatrically healthy women to commission errors in two variants of the CPT - with and without interstimulus distracters. Controlling for age and IQ, testosterone did not relate to rates of correct detections in either task, but correlated positively with commission errors in the distracter CPT variant. In light of the fact previous studies find commission errors on the CPT are associated with impulsivity, the results of this study support a positive relationship between testosterone and impulsivity. PMID- 11399315 TI - The differential effects of intravenously administered 8-OH-DPAT on operant food intake in satiated and food-deprived pigs are mediated by central 5-HT(1A) receptors. AB - It has previously been shown that the intravenous administration of the 5-HT(1A) receptor agonist, 8-hydroxy-2-(di-n-propylamino) tetralin (8-OH-DPAT), increases food intake in satiated pigs and decreases food intake in fasted pigs. The present experiments were conducted to investigate the effects of central administration of the 5-HT(1A) receptor antagonist, N-[2-[4-(2-methoxyphenyl)-1 piperazinyl]-N-2-pyridinyl-cyclohexane carbox-amide maleate (WAY 100635), on the stimulant and depressant effects of 8-OH-DPAT on operant food intake in satiated and hungry pigs. In Experiment 1, 8-OH-DPAT (25 microg/kg) produced an increase in operant feeding during the first 30 min following intravenous administration to satiated pigs. The 8-OH-DPAT-induced hyperphagia was completely abolished by pretreatment with WAY 100635 (0.3 mg) administered by intracerebroventricular injection. In Experiment 2, 8-OH-DPAT (25 microg/kg) administered intravenously 15 min prior to the onset of feeding in pigs that had been fasted for 22.5 h produced a decrease in operant food intake, which was most apparent during the first 30 min of the feeding period. The hypophagic effect was completely abolished by pretreatment with WAY 100635 (0.3 mg icv) administered 30 min before the start of the feeding period. In both experiments, WAY 100635 (0.3 mg icv) did not have any significant effects on feeding. The results of the present study extend previous results in the pig and show that both the hyperphagic and the hypophagic effects of 8-OH-DPAT in satiated and fasted pigs, respectively, are mediated by central 5-HT(1A) receptors. PMID- 11399316 TI - Leptin reverses sucrose-conditioned place preference in food-restricted rats. AB - Previous studies have suggested that food restriction can modify performance in the conditioned place preference (CPP) paradigm. In the present study, we tested the hypotheses that food restriction would enhance the development of a CPP to low-calorie sucrose pellets and that peripheral leptin replacement in food restricted animals would reverse this effect. Using a range of 45-mg sucrose pellets (0-15 pellets) as a reward, we observed that a significant place preference was conditioned in food-restricted, but not ad libitum-fed rats. This CPP was reversed either by treatment of food-restricted rats with the dopamine receptor antagonist alpha-flupenthixol (200 microg/kg ip) during the training protocol or by chronic subcutaneous replacement of leptin (125 microg/kg/day) that attenuated the food restriction-induced decrease of circulating leptin. We conclude that dopaminergic signaling and the fall of plasma leptin concentrations contribute to the CPP of food-restricted rats. This finding suggests that in addition to metabolic adaptations, hypoleptinemia results in behavioral adaptations during states of energy deprivation. PMID- 11399317 TI - Individual variation in the relation between body temperature and energy expenditure in response to elevated ambient temperature. AB - The question we address here is whether a mild increase in environmental temperature affects body temperature and energy expenditure, focusing on the individual variation in the relation between energy expenditure and body temperature. We studied eight normal weight healthy females, 48 h at an ambient temperature of 22 degrees C, and 48 h at 27 degrees C. Energy expenditure (EE) was measured in a respiration chamber. Subjects' skin temperature was measured continuously from 8:00 a.m. until 12:00 p.m.: forehead, infraclavicular zone, thigh, hand, and foot. Core temperature was determined tympanically. Body composition was determined by under water weighing. Exposure to 27 degrees C caused a significant increase in body temperature (both skin and core), a decrease in temperature gradients, and a decrease in energy expenditure. At 27 degrees C 24 h EE, adjusted for body composition, was significantly related to body tympanic temperature. The decrease in 24 h EE, at 27 degrees C ambient temperature, was significantly, negatively related to the increase in T(tym), indicating individual responses in adaptation to elevated ambient temperature. Changes in temperature gradient (comparing 27 degrees C with 22 degrees C) were negatively related to changes in EE. This shows that individuals differ in their response to an increase in environmental temperature regarding the relative contribution of insulative or metabolic adjustments. PMID- 11399318 TI - Effects of feeding corticosterone and housing conditions on feather pecking in laying hens (Gallus gallus domesticus). AB - Feather pecking (FP) and cannibalism in laying hens are disadvantageous to the well-being of the birds. We investigated whether stress could be proposed as a trigger for the development of this abnormal behavior. From Week 11 to 19 after hatching, 16 groups of 15 or 16 hens (white Lohman Selected Leghorn hybrids) were kept in pens with or without foraging material (litter) and fed a diet containing corticosterone (C, 1.5 mg/bird/day) or no C. Birds fed on C had reduced values for weight gain and egg production, prolonged tonic immobility (TI), higher heterophil/lymphocyte ratios (H/L) and higher serum C concentrations. On litter, C-fed birds developed significantly higher rates of FP than when not fed C. However, birds kept on slats also developed high rates of FP but without being fed C. Feeding C to these birds did not significantly further increase the rates of FP. We concluded that FP may develop as a response to increased blood C concentrations, but that housing conditions restricted in relation to foraging material, may as well induce FP in the absence of increased C levels. PMID- 11399319 TI - Human natural killer cell activating receptors. AB - Natural killer (NK) cells were poorly characterized until 10 years ago and few molecules expressed on their cell surface were known. Now the situation has changed dramatically, since a plethora of receptors characterized by opposite functions have been functionally and molecularly defined. NK cells express clonally distributed inhibitory receptors specific for different groups of HLA class I alleles, thus protecting normal cells from NK-mediated lysis. On the contrary, various activating receptors are involved in triggering of NK-mediated natural cytotoxicity. Their engagement induces human NK cells to kill target cells that are either HLA class I-negative or -deficient. Here a brief description of the activating receptors and coreceptor and of their ligand(s) is given. PMID- 11399320 TI - Antibody repertoire development in fetal and neonatal piglets. V. VDJ gene chimeras resembling gene conversion products are generated at high frequency by PCR in vitro. AB - The recovery of VDJ rearrangement is most often accomplished by PCR amplification of DNA extracted from mixtures of B-cells. Using this procedure in swine, VDJs containing chimeric V(H) genes that resemble gene-conversion products, are frequently encountered. To examine whether these chimeras could be the result of PCR artifacts, we used different combinations of swine VDJ templates, each having unique CDR1, CDR2 and D(H) segments, to generate >2600 clones. Using equal amounts of two templates and 30 cycles of PCR, up to 45% of the resultant clones were VDJ chimeras. The frequency of chimeras was independent of the specific VDJ template and the chimeras were generated regardless of whether Taq-, Pfu- or mixtures of Taq- and Pfu-polymerases were employed or whether PCR extension time was prolonged six-fold. The frequency of generating chimeras was dependent on the ratio of the two target DNAs although even ratios approximately 1:10 generated approximately 10% chimeric VDJs. Chimeras could be generated using only 10 cycles of PCR or using the initial template DNAs diluted as much as 1:10000. Of the 279 chimeric VDJs generated, 61% of the crossovers occurred in FR3, 21% in FR2 and 18% in both FR2 and FR3. We interpret these results to mean that in vivo gene conversion in this species can only be unambiguously proven when the VDJs from individual B-cells are bearing a single VDJ rearrangement amplified and sequenced or when VDJs are cloned without the use of PCR. PMID- 11399321 TI - Dissection and optimization of immune effector functions of humanized anti ganglioside GM2 monoclonal antibody. AB - A mouse/human chimeric monoclonal antibody (MAb) KM966, specific for the cell surface tumor antigen ganglioside GM2, was humanized by the complementarity determining regions (CDRs) grafting method. Not only the amino acid residues in the CDRs but also several in the framework regions (FRs) were changed from the human to the murine residues. A humanized variant, huKM796H/Lm-28, containing eight and five amino acid alterations in variable light (VL) and variable heavy (VH) FRs, respectively, showed a 9-fold reduction in complement-dependent cytotoxicity (CDC) compared to the chimeric KM966, despite tight antigen binding and potent antibody-dependent cellular cytotoxicity (ADCC). Several additional variants were subsequently constructed to improve the CDC of the antibody. One of the variants, designated KM8969, which differs by three amino acids, exhibited a CDC within 3-fold of the chimeric KM966. In addition, humanized KM8969 bound GM2 antigen 1.25-fold more tightly than the chimeric KM966 and showed 5-fold higher ADCC than the chimeric KM966. These results clearly show that the humanized KM8969, having the optimized immune effector functions and theoretically minimal immunogenicity, is an ideal candidate to test the effectiveness of anti-GM2 MAb in human cancer therapy. Taken together, the results obtained here indicate that the ADCC and CDC of an antibody can be dissected independently via engineering of the antibody variable region. PMID- 11399322 TI - Heterogeneity of O-glycosylation in the hinge region of human IgA1. AB - Matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry was applied to studies of the molecular heterogeneity of desialylated human IgA1 hinge region glycopeptides released with two IgA1 proteases. Typically, the hinge region of an alpha1 chain contains three to five O-linked glycan chains. Variants of the hinge region peptides released from IgA1(Kni) myeloma protein carrying 0, 1, 2, or 3 GalNAc residues were observed in the mass spectra as well as the nonglycosylated peptide. Variable numbers of Gal residues indicated additional heterogeneity in O-glycosylation of IgA1. In the hinge region preparation from normal human serum IgA1, glycopeptides carrying 2, 3, 4, or 5 GalNAc residues with variable numbers of Gal residues were detected. In conclusion, our new approach using the site-specific cleavage with two IgA1 proteases allowed precise and sensitive MALDI-TOF mass spectrometric analysis of O-glycosylation heterogeneity in IgA1 hinge region. PMID- 11399323 TI - Differential interaction of Cbl with Grb2 and CrkL in CD2-mediated NK cell activation. AB - Natural killer (NK) cells participate in both innate and adoptive immunity by their prompt secretion of cytokines and by their ability to lyse virally infected cells or tumor cells. CD2 is surface glycoprotein receptors and crucial for NK cell activation. However, molecular events involved in CD2-mediated NK cell activation have not been fully elucidated. Cbl-Grb2 and Cbl-CrkL interactions have been implicated in T cell and B cell receptor, and cytokine receptor signaling. Here we analyzed tyrosine phosphorylation and interactions of Cbl with adapter proteins, Grb2 and CrkL, in NK3.3 cells. CD2 crosslinking results in the marked tyrosine phosphorylation of Cbl in an antibody concentration- and time dependent manner. Immunodepletion studies reveal that Grb2-associated tyrosine phosphorylated p120 kDa protein is Cbl. In vitro binding studies using GST-fusion proteins demonstrate that Cbl constitutively associates with the SH3 domains of Grb2, with a preference for the amino-terminal domain. In addition, we demonstrate that CrkL associates with a large portion of tyrosine phosphorylated Cbl after CD2 stimulation of NK3.3 cells. In contrast to constitutive Cbl association with Grb2, tyrosine phosphorylated Cbl interacts with CrkL via its SH2 domain only after CD2 stimulation. Although the precise roles of interactions of Cbl with Grb2 and CrkL in NK cell activation remains to be elucidated, their tyrosine phosphorylation, in addition to the multiple protein interactions described here, strongly suggest that interactions of Cbl with Grb2 and CrkL may play pivotal roles in CD2-mediated NK cell activation. PMID- 11399324 TI - Streptabody, a high avidity molecule made by tetramerization of in vivo biotinylated, phage display-selected scFv fragments on streptavidin. AB - Phage display is a powerful method of isolating of antibody fragments from highly diverse naive human antibody repertoires. However, the affinity of the selected antibodies is usually low and current methods of affinity maturation are complex and time-consuming. In this paper, we describe an easy way to increase the functional affinity (avidity) of single chain variable fragments (scFvs) by tetramerization on streptavidin, following their site-specific biotinylation by the enzyme BirA. Expression vectors have been constructed that enable addition of the 15 amino acid biotin acceptor domain (BAD) on selected scFvs. Different domains were cloned at the C-terminus of scFv in the following order: a semi rigid hinge region (of 16 residues), the BAD, and a histidine tail. Two such recombinant scFvs directed against the carcinoembryonic antigen (CEA) were previously selected from human non-immune and murine immune phage display libraries. The scFvs were first synthesized in Escherichia coli carrying the plasmid encoding the BirA enzyme, and then purified from the cytoplasmic extracts by Ni-NTA affinity chromatography. Purified biotinylated scFvs were tetramerized on the streptavidin molecule to create a streptabody (StAb). The avidity of various forms of anti-CEA StAbs, tested on purified CEA by competitive assays and surface plasmon resonance showed an increase of more than one log, as compared with the scFv monomer counterparts. Furthermore, the percentage of direct binding of 125I-labeled StAb or monomeric scFv on CEA-Sepharose beads and on CEA expressing cells showed a dramatic increase for the tetramerized scFv (>80%), as compared with the monomeric scFv (<20%). Interestingly, the percentage binding of 125I-labeled anti-CEA StAbs to CEA-expressing colon carcinoma cells was definitely higher (>80%) than that obtained with a reference high affinity murine anti-CEA mAb (30%). Another advantage of using scFvs in a StAb format was demonstrated by Western blot analysis, where tetramerized anti-CEA scFv could detect a small quantity of CEA at a concentration 100-fold lower than the monomeric scFv. PMID- 11399325 TI - Selective deficits in attentional performance on the 5-choice serial reaction time task following pedunculopontine tegmental nucleus lesions. AB - Sustained attention requires the integrity of basal forebrain cholinergic systems. The pedunculopontine tegmental nucleus (PPTg) has direct and indirect connections (via the thalamus) with the basal forebrain, suggesting that the PPTg may also play an important role in attentional processes. We examined this hypothesis by testing the effects of PPTg lesions in rats on performance in the 5 choice serial reaction time test. Bilateral lesions reduced accuracy, increased errors of omission, and increased the latency to correct responses. The deficits were more severe when neuronal damage was bilateral and concentrated in the posterior PPTg. Attentional demands of the task were increased by decreasing the stimulus duration, the stimulus brightness, or the inter-trial interval, and by introducing random bursts of white noise. These challenges impaired performance of all animals, but the magnitude of deficit was increased in the lesioned group. Conversely, lesion-induced deficits were partially alleviated when the attentional demands of the task were reduced. This pattern of results suggests that PPTg lesions produce a global deficit in attention, rather than a specific impairment in one process. The PPTg may control attentional processes through its direct projections to the forebrain cholinergic system or, indirectly, through activation of thalamocortical projections. PMID- 11399326 TI - Sensitivity of cortical movement representations to motor experience: evidence that skill learning but not strength training induces cortical reorganization. AB - The topography of forelimb movement representations within the rat motor cortex was examined following forelimb strength training. Adult male rats were allocated to either a Power Reaching, Control Reaching or Non-Reaching Condition. Power Reaching rats were trained to grasp and break progressively larger bundles of dried pasta strands with their preferred forelimb. Control Reaching animals were trained to break a single pasta strand and Non-Reaching animals were not trained. Power Reaching animals exhibited a progressive increase in the maximal size of the pasta bundle that could be retrieved during a 30-day training period. Kinematic analyses showed that this improvement was not due to a change in reaching strategy. Intracortical microelectrode stimulation was used to derive maps of forelimb movement representations within the motor cortex of all animals following training. In comparison to Non-Reaching animals, both Power Reaching and Control Reaching animals exhibited a significant increase in the proportion of motor cortex occupied by distal forelimb movement representations (wrist/digit) and a decrease in the proportion of proximal representations (elbow/shoulder). These results demonstrate that the development of skilled forelimb movements, but not increased forelimb strength, was associated with a reorganization of forelimb movement representations within motor cortex. PMID- 11399327 TI - Role of glutamate receptors in the nucleus accumbens on behavioural responses to novel conflictive and non-conflictive environments in the rat. AB - The possible role of glutamic acid locally applied into the nucleus accumbens on exploratory behaviours measured in 'conflictive' and 'non-conflictive' environments was studied in adult male rats. As a model of conflictive environment, the elevated asymmetric-plus maze (APM) was used. As a model of a non-conflictive environment, a modified holeboard enriched with an object (OVM) was used. In order to characterize the possible glutamic acid receptors involved, the following antagonists were also used: AP3 (antagonist of the metabotropic glutamic acid receptor), AP7 (antagonist of NMDA glutamic acid receptor, and CNQX (antagonists of kainate/AMPA glutamic acid receptor). Results showed that injection of glutamic acid into the nucleus accumbens induced in the APM a decrease of exploration and an increase of the permanency score (non-exploratory behaviours) of the 'High and Low wall' arm. However, in the 'Two High Walls' arm, glutamic acid decreased permanency. In the OVM, no major changes in the motor activity were observed with glutamic acid. Nevertheless, the vertical activity (an index of rearing) and head-dipping were inhibited by the amino-acid treatment. In the APM, the decrease of exploration induced by glutamic acid was blocked by all three receptor antagonists. In the non-exploratory behaviours, the facilitatory effect observed in the 'High and Low walls' arm was blocked only by AP7 and CNQX. The inhibitory action of glutamic acid on the permanency score in the 'Two High Walls' arm was not blocked by the receptors antagonists. In the OVM, AP7 and CNQX were effective in blocking the inhibition of glutamic acid on the vertical activity, but in head-dipping, only AP3 and CNQX were able to block the effect of the amino acid on this behaviour. In conclusion, the present results are compatible with the concept that glutamatergic input fibres to the nucleus accumbens modulate the expression of exploratory behaviour induced by novelty in conflictive and non-conflictive conditions. PMID- 11399328 TI - REM sleep enhancement due to rhythmical auditory stimulation in the rat. AB - From a physiological viewpoint, REM sleep (REMS) is a period during which homeostatic physiological regulations are impaired. In the rat, REMS occurs in two forms respectively characterized by episodes separated by long intervals (single REMS episodes) and by episodes which have short intervals and occur in sequences (REMS clusters). Since the partition of REMS in the form of either single or clustered episodes may reveal how the REMS drive and body homeostatic processes interact in the control of REMS occurrence, we have used this approach to clarify the effects of the rhythmical delivery of an auditory stimulus (1000 Hz, 63 or 88 dB, 50 ms, every 20 s), which has been previously observed by different authors to enhance REMS in the absence of a previous sleep deprivation. Stimuli were delivered to pairs of animals and triggered by the occurrence of REMS in one rat (REMS-selective stimulation), whilst the other animal received the same stimulus irrespectively of the stage of the wake-sleep cycle (REMS unselective stimulation). The results showed that the REMS-selective stimulation did not change the overall amount of REMS, since an increase in the occurrence of REMS clusters was concomitant with a decrease in the occurrence of single REMS episodes. In contrast, under the REMS-unselective stimulation, the total amount of REMS was increased during the second day of stimulation through an increase in the duration of both types of REMS episodes. Since during the REMS-unselective stimulation 87% of the stimuli fell outside REMS (i.e., during the REMS interval), the results show that the occurrence of REMS is more consistently affected when the stimuli are delivered in a period during which homeostatic physiological regulations are fully operant. PMID- 11399329 TI - Neurons in rat medial prefrontal cortex show anticipatory rate changes to predictable differential rewards in a spatial memory task. AB - The present study electrophysiologically examined the contribution of prelimbic and infralimbic neurons in the medial prefrontal cortex (mPFC) to integration of reward and spatial information while rats performed multiple memory trials on a differentially rewarded eight arm radial maze. Alternate arms consistently held one of two different reward amounts. Similar to previous examinations of the rat mPFC, few cells showed discrete place fields or altered firing during a delay period. The most common behavioral correlate was a change in neuronal firing rate prior to reward acquisition at arm ends. A small number of reward-related cells differentiated between high and low reward arms. The presence of neurons that anticipate expected reward consequences based on information about the spatial environment is consistent with the hypothesis that the mPFC is part of a neural system which merges spatial information with its motivational significance. PMID- 11399330 TI - Neurobehavioural defects in adult mice neonatally exposed to nicotine: changes in nicotine-induced behaviour and maze learning performance. AB - Neonatal exposure to low doses of nicotine has been shown to disturb the development of low-affinity nicotinic binding sites in the cerebral cortex and to elicit a deviant behavioural response to nicotine in adult mice. In this study, 10-day-old male NMRI mice were exposed to one of three different doses of nicotine (3.3, 33, or 66 microg nicotine-base/kg body wt.) s.c. twice daily on 5 consecutive days to study dose-response effects of nicotine on adult spontaneous and nicotine-induced motor behaviour. The nicotine-induced behaviour test revealed a hypoactive response to nicotine in 4-month-old mice neonatally exposed to 33 or 66 microg nicotine-base, whereas the response to nicotine in control animals and mice exposed to 3.3 microg nicotine-base was an increased activity. Learning and memory functions were also investigated in adult animals neonatally exposed to 66 microg nicotine-base/kg body wt. in the same manner, in the Morris water maze and in the Radial arm maze. In the swim maze and the Radial arm maze tests, no significant differences were observed between nicotine-treated and control animals at the age of 4 months. At 7 months, however, a significant difference in performance was evident, indicating a time-response/time-dependent effect. Furthermore, it was shown that in mice exposed neonatally to a nicotine dose known to inhibit the development of the nicotinic low affinity-binding site (LA), the response to nicotine could not cause any increase in spontaneous motor activity as seen in controls. PMID- 11399331 TI - Studies on a model of long term alcohol drinking. AB - This study investigated the effects of a dihydropyridine calcium channel antagonist in a free drinking model previously reported to show increased and 'uncontrolled' drinking. The results showed, rather than the previously reported increase in consumption, a gradual decrease in alcohol intake over 9-18 months. When the alcohol was withdrawn from one group of rats after 55 weeks free choice, the animals showed no behavioural signs of physical withdrawal, but they did demonstrate the expected elevated ethanol intake on reintroduction to ethanol after 2 weeks abstinence. A second group of rats were given 62 weeks free choice access to ethanol in groups of four, then transferred to single housing and baseline drinking levels established. Intraperitoneal injections of nimodipine 5 mg/kg, 20 mg/kg or Tween vehicle were then given once daily. Nimodipine had no effect on ethanol intake of animals with continuous access to ethanol, or of those animals withdrawn from ethanol and then reintroduced after 2 weeks of abstinence. However the unexpectedly low alcohol intake may have prevented any effects of nimodipine being seen. PMID- 11399332 TI - Constraints during bimanual coordination: the role of direction in relation to amplitude and force requirements. AB - The present study addressed the status of spatial encoding during a bimanual task paradigm. This was based on the premise that patterns of contralateral interference during bimanual coordination provide a window into those movement parameters that are primarily encoded within the central nervous system. Results showed that both direction and amplitude were subject to (bilateral) interference when different specifications were to be generated simultaneously for each limb. Directional interference was found to be partially independent of the amount and pattern of underlying muscle activation, suggesting that direction is encoded at a rather abstract level in the central nervous system. The findings are consistent with single-cell recording studies that have pointed to the role of directional tuning in various brain areas. Moreover, the findings suggest that spatial parameters of movement constrain the coordination of limb movements in addition to temporal parameters. PMID- 11399333 TI - Evoked and oscillatory neuromagnetic responses to sniffing odor in human subjects. AB - We studied neuromagnetic responses to sniffing air and odor of lemon, lavender or soy sauce in two human subjects. Simple averaging of the data with respect to the onsets of the sniffs yielded odor-specific evoked responses over the right cortex with peak latency about 350 ms after the onsets. When the data were passed through band-pass filters centered around 48 Hz and of 320 ms in duration, rectified and then averaged, they yielded peaks, indicating transient oscillations, over the right frontal cortex with longer latency. We found such peaks in only 9 out of 26 experiments of sniffing odor, however. PMID- 11399334 TI - Medical malpractice, murder and the academic community: trouble ahead. AB - The morale of the medical professional is at an historic low in many countries. The recent case of a UK general practitioner being convicted of being a mass murderer, combined with increasing criticism of medical negligence and malpractice and an excessive influence of the large pharmaceutical companies has lead to the perception that the profession is under siege. Our professional leadership have not had sufficient public awareness to allay these concerns, and the resulting dip in morale is fast turning into a dangerous rout. We must review what has lead to this situation and what we should be doing now to put it right.Clinical research is under attack and the motives and ethics of large pharmaceutical company sponsorships of clinical trials is under increasing question. At this time there is a risk that medicine, and academic medicine in particular, will lose its attractiveness and the pace of achievements we have seen and benefited from in the last 2 decades may slow. The public debate should move on, it should move on to evaluate how much it would cost to reduce medical error rates to an acceptable level (to stop them altogether is impossible). It should move on to how we can get clinical trials designed and paid for by the public purse rather than merely grumbling that pharmaceutical companies take too much control of trials that they almost alone now appear to be sponsoring. And we should move on to debate about the role and status of the medical profession in the modern era. We can no longer do our best in secret and expect the public to trust us unquestioningly. The public wants and needs to be involved in our decision-making problems and errors. Only through informed debate will we improve health for the while population, now and in the future. PMID- 11399335 TI - Beta-blocking in heart failure patients. Balancing the evidence. AB - Evidence for the effectiveness of beta-blockers in the management of patients with heart failure is now compelling with a database of over 13000 patients enrolled in randomised prospective placebo-controlled clinical trials. However this therapy remains vastly underused in clinical practice. The different points challenging the widespread use beta blockade agents in the routine treatment in heart failure are presented and discussed. After a review of the potential mechanism hypothesised behind the benefits of beta-blockers in heart failure, the controversial effects on the haemodynamics, exercise tolerance, hospitalisation and mortality are underlined. PMID- 11399336 TI - Another view of myocardial hibernation. AB - This manuscript brings together three newer concepts: myocardial hibernation, heterogeneity in myocardial blood flow and oxidative metabolism, and effects of hibernating animal serum on non-hibernators. Myocardial hibernation is viewed as a protective mechanism that helps to maintain myocardial integrity and viability by down-regulating contractile function as an adaptation to reduced blood flow. Myocardial flow is considerably heterogeneous. Consequently, oxygen supply to the myocardium is also heterogeneous. Many lines of evidence show a close correlation between regional flow and regional metabolism. In low-flow/low-metabolism areas, myocardial function must be reduced, since the myocardium would otherwise undergo necrosis. Because no regional histological differences exist, the pattern of heterogeneity seems to shift over time. Hence, we hypothesize that such very regional hibernation presents an evolutionary, protective mechanism, permitting subsequent myocardial areas to rest within the ceaselessly working heart. We also hypothesize that a similar mechanism ensures the down-regulation of function following myocardial ischemia in order to induce myocardial hibernation on a broader level. Surprisingly, a substance (opioid in nature) contained in hibernator serum both induced hibernation-like state in non-hibernators and suppressed myocardial oxygen consumption. Thus, we lastly hypothesize that myocardial hibernation is a remnant of the early stages of evolution and is closer to physiological hibernation than traditionally viewed. PMID- 11399337 TI - Endothelin release: a marker for the severity of exercise-induced ischemia. AB - Elevated endothelin-1 levels were found in exercise-induced ischemia. To our knowledge, no attempt has been made to correlate the presence of the elevated endothelin levels with severity of ischemia. In the present study, we attempt to correlate the severity of ischemia, quantified using a bullseye visual display of scintigraphic ischemic score on SPECT technetium 99m sestamibi, with presence of endothelin at peak exercise. Twenty seven patients with documented effort angina were included in the study. All subjects underwent stress technetium 99m sestamibi and ischemic scores were evaluated on the polar map image divided into 13 segments. Endothelin levels were measured at baseline and at peak exercise. In 13 patients with elevated endothelin-1 levels, there were 88 ischemic segments, 26 of which showed severe ischemia. In the remaining 14 patients with no endothelin-1 elevation, there were 80 ischemic segments of which only four showed severe ischemia. The amount of severe ischemic segments per patient was 2+/-2.2 in the elevated endothelin positive group and 0.28+/-0.6 in the endothelin negative (P< or =0.011). When looking at ischemia as detected by sestamibi SPECT as a gold standard, we found that of 331 segments, 168 were without ischemia, 143 had mild to moderate ischemia, and 30 has severe ischemia. Endothelin levels were 1.2+/-0.6, 2.2+/-0.5, 6.2+/-0.7 pg/l (P<0.01), respectively. Thus, endothelin-1 is a marker of severity of ischemia rather than ischemia itself, and as such, may have prognostic value. PMID- 11399338 TI - Circulating N-terminal brain natriuretic peptide precursor and endothelin levels in patients with syndrome X and left bundle branch block with preserved systolic function. AB - BACKGROUND: Deterioration of left ventricular function during follow-up was reported in some patients with syndrome X and concomitant left bundle branch block. The patients with syndrome X and left bundle branch block has been frequently presented with elevated Endothelin-1 (ET-1) level while brain natriuretic peptide (BNP) (a sensitive marker of left ventricular dysfunction) has not been measured in patients with syndrome X. METHODS: The purpose of the present study was to assess left ventricular diastolic function, levels of N terminal Brain Natriuretic Peptide (NT-proBNP) precursor and biochemical parameters of endothelial function in patients with syndrome X complicated by left bundle branch block but preserved left ventricular systolic function (group A, n=8). The echocardiographic and neurohormonal measures in these patients were compared to those in patients with syndrome X without left bundle branch block (group B, n=13), and controls (group C, n=15). RESULTS: At rest and after exercise the serum concentration of NT-proBNP was significantly higher in group A than in the controls (at rest: 232+/-96 vs. 133+/-23 fmol/ml, P=0.03; after exercise: 313+/-96 vs. 180+/-33 fmol/ml, P=0.02). The highest concentration of endothelin-1 was also found in group A, being significantly higher than in the controls (6.81 vs. 4.52 pg/ml, P<0.05). Mitral flow abnormalities were detected in left bundle branch block patients. Accordingly, the lowest E/A ratio was in group A and it differed significantly from that in group C (0.85 vs. 1.1, P<0.05). E/A ratio inversely correlated with plasma NT-proBNP concentration in patients with left bundle branch block (r=-0.48, P=0.02). CONCLUSIONS: Elevated NT-proBNP and endothelin-1 plasma concentrations were demonstrated in patients with syndrome X complicated by left bundle branch block even when left ventricular systolic function was still preserved. In this subgroup the magnitude of left ventricular diastolic dysfunction correlated with the increase of BNP level which reflects neurohormonal activation. PMID- 11399339 TI - Effect of beta-blockade on heart rate variability in decompensated heart failure. AB - BACKGROUND: One of the putative mechanisms for the salutary effects of beta blockers in patients with congestive heart failure is their ability to improve autonomic dysfunction. However, patients with profound neurohumoral abnormalities derive little survival benefit from beta-blockers. The purpose of the current study was to evaluate the effect of beta-blockers on heart rate variability in decompensated heart failure. METHODS: Time and frequency domain heart rate variability indices were obtained from 24-h Holter recordings and compared to assess the role of beta-blockade in 199 patients (mean age 60+/-14 years [range 21 to 87]) with decompensated heart failure (New York Heart Association functional class III [66%] and IV [34%]). RESULTS: All heart rate variability indices were markedly suppressed but were substantially higher in patients who were on beta-blockers. Time domain measures of parasympathetic cardiac activity, the percentage of RR intervals with >50 ms variation (4.9+/-0.6 vs. 7.7+/-1.2%, P=0.006) and the square root of mean squared differences of successive RR intervals (22.7+/-2.0 vs. 31.6+/-4.1 ms, P=0.004), were higher in the beta blocker group. Spectral analysis revealed that the total power and the ultra low frequency power were significantly higher in patients on beta-blockers (82% and 59%, respectively). The high frequency power, a spectral index of parasympathetic modulation, was 41% higher in the beta-blocker group (121+/-25 vs. 171+/-27 ms(2), P=0.02). Multiple linear regression, adjusted for clinical parameters and drug therapies, revealed a strong positive relationship between beta-blockade and higher values of time and frequency domain measures. The mean number of ventricular tachycardia episodes were significantly lower in patients on beta blocker therapy (3.6+/-1.5 vs. 19.0+/-5.3, P=0.04). CONCLUSIONS: beta-blockers improve the impaired cardiac autonomic regulation during high sympathetic stress of decompensated heart failure. PMID- 11399340 TI - Outcome of patients with post acute myocardial infarction ischemia hospitalized in intensive cardiac care units with and without on-site catheterization laboratories. AB - BACKGROUND: This study addresses the impact of availability of on-site catheterization laboratories on the 1-year survival of patients with post-acute myocardial infarction ischemia (P-AMI-I), a high-risk subgroup of AMI patients. METHODS: A prospective 5 month national survey was conducted in 1996 in all operating intensive care units (ICCUs) in Israel (N=26) and included 2377 patients. Four hundred and three (17%) had P-AMI-I, 317 of them were admitted to 18 ICCUs with on-site catheterization laboratories (CATH+) and 86 patients to 8 ICCUs without such facilities (CATH-). A retrospective analysis was performed comparing the in-hospital course and 7 day, 1 month and 1 year mortality data of CATH+ vs. CATH- patients. RESULTS: Patient characteristics in both groups were similar with regard to age, gender AMI location, risk factors, hemodynamic parameters on admission and rate of thrombolytic therapy. Of patients in CATH+, 79% were catheterized before hospital discharge vs. 42% in CATH- (P<0.0001), 45 vs. 15% had PTCA (P<0.0001) and 19 vs. 9% had CABG (P<0.05). At 30 days, patients in CATH+ still had significantly more revascularization procedures (71 vs. 48%, P<0.001). Patients hospitalized in ICCUs with CATH+ and CATH- facilities had similar cardiac mortality rates at 7 days (2.0 vs. 2.3%), 30 days (5.7 vs. 4.7%) and at 1 year (7.6 vs. 7.0%). CONCLUSIONS: Despite a more invasive strategy used during the index hospitalization of patients with P-AMI-I hospitalized in CATH+ ICCUs, their survival was similar to CATH- patients at 7 days, 30 days and at 1 year follow-up. PMID- 11399341 TI - Activation of circulating platelets and platelet response to activating agents in children with cyanotic congenital heart disease: their relevance to palliative systemic-pulmonary shunt. AB - Abnormal platelet function has been hypothesised to play a role in the haemostatic abnormalities in cyanotic congenital heart disease (CCHD) patients. Using whole blood flow cytometry we found that platelets from cyanotic patients were hyperreactive and we related such hyperreactivity directly to young age, unoperated state, high haematocrit, reduced saturation with oxygen and low platelet count. Circulating platelets from CCHD children showed significantly enhanced P-selectin expression (P<0.004) and remained more reactive to 0.2 IU/ml thrombin, 1-8 microM TRAP and 2-4 microM ADP (P<0.04), especially in younger (0-3 year-olds) patients. Such a platelet 'priming' largely concerned CCHD children who were not subjected to modified Blalock-Taussig shunts in the past (non-MBTS). Only non-MBTS cyanotic children, but not MBTS-operated patients, showed significantly higher platelet reactivity compared to controls in response to ADP or 1 microM TRAP with respect to P-selectin expression (p<0.05) and in response to all examined agonists with respect to GPIb expression (P<0.045). The enhanced P-selection expression in MBTS-operated CCHD children and reduced GPIb expression in non-MBTS patients, especially in younger patients, were positively associated with the occurrence of the polymorphic variant Pl(A2) of platelet membrane glycoprotein IIIa gene. Altered blood morphology parameters (elevated RBC, Hb, Hct and MCHC, for all P<0.0005) in CCHD children correlated with the enhanced degranulation of circulating blood platelets and their hyperreactivity in response to some agonists (P<0.05). Overall, our data encourage the reasoning that circulating platelets are remarkably hyperreactive in non-MBTS cyanotic children, which are at higher risk to often encounter platelets activation in circulation. It seems unlikely that the apparently unchanged platelet reactivity in MBTS-operated children is due to the advantageous effects of the shunt, since these patients showed neither altered haematological parameters nor improved oxygen carrying capacity. Otherwise, it may rather result from more frequent episodes of platelet degranulation and preactivation in the past, and/or post operative enhanced platelet consumption. PMID- 11399342 TI - Circadian variation of heart rate variability and the rate of autonomic change in the morning hours in healthy subjects and angina patients. AB - BACKGROUND: Incidence of sudden cardiac death peaks during the early morning hours when there is a rapid withdrawal of vagal and an increase of sympathetic tone. The rate of autonomic change could be of prognostic importance. PATIENTS AND METHODS: A total of 65 patients with angina pectoris, free from other diseases and drug free, were Holter monitored for 24 h. A total of 30 patients were also monitored on isosorbide-5-mononitrate (IS-5-MN) and on metoprolol respectively. A total of 33 age-matched healthy subjects served as controls. Spectral components of heart rate variability (HRV) were analysed hourly, with special reference to the rapid changes of autonomic tone during the night and early morning hours. Circadian variation was assessed in two ways: (1) Mean HRV day (8 a.m.-8 p.m.) and night (0-5 a.m.) were compared. (2) For the morning/night hours (0-10 a.m.), individual hourly values for max. and min. HRV, the difference max.-min. (gradient), the rate of change per hour between max. and min. (velocity) and the largest difference between two consecutive hours (max. velocity) were recorded and the mean value for the group calculated. RESULTS: During the night/morning hours, healthy controls demonstrated faster HF max. velocity (P=0.002) and higher HF gradient (P=0.011) than angina patients. Metoprolol and IS-5-MN increased the HF gradient (P=0.008 and P=0.003, respectively), and metoprolol tended to increase the max. velocity (P=0.02). Metoprolol substantially decreased the LF/HF gradient (P=0.001), velocity (P=0.008) and max. velocity (P=0.0001). CONCLUSION: Rapid vagal withdrawal seemed to be a sign of a healthy autonomic nervous system in the control group but was significantly slower in angina patients. IS-5-MN and metoprolol tended to normalise vagal withdrawal and metoprolol slowed down the rapid increase in sympathetic predominance in the morning in patients. PMID- 11399343 TI - Transcatheter occlusion of the patent ductus arteriosus with a single device technique: comparison between the Cook detachable coil and the Rashkind umbrella device. AB - Transcatheter coil occlusion of the patent ductus arteriosus (PDA) has become the interventional treatment option of choice. Immediate occlusion of any residual shunting results in excellent closure rates, but frequently requires multiple coil deployment. AIMS: To assess the efficacy and limitations of single Cook detachable coil PDA closure compared to a preceding series of Rashkind umbrella procedures. METHODS AND RESULTS: Between 1990 and 1999, transcatheter occlusion of a small (<2 mm; n=45) or moderate-sized (2-4 mm; n=47) PDA was successfully attempted in 90/92 consecutive patients (mean age 6+/-4.8 years) with a coil (39/41) or Rashkind device (51/51). Immediate angiographic closure rates for both devices were low, although better for small (54-68%) than moderate ducts (7-22%, P<0.01). A 2-year echocardiographic closure rate of small ducts increased to 92% for the coil group versus 95% for the Rashkind group. By that time, moderate sized ducts were only occluded in 64% with the coil and 54% with the Rashkind device. A visible residual shunt at post-implant angiography in moderate ducts was associated with a high incidence (59%) of long-term echocardiographic shunt patency and a need for repeat interventions for audible residual shunts (32%). CONCLUSIONS: Single coil transcatheter occlusion is the treatment of choice for the small duct as most residual shunts will resolve spontaneously. However, long term shunt persistence after single coil deployment in moderate sized ducts is as frequent as with the Rashkind device. A primary multiple coil approach is advocated if the postcoil aortogram shows residual ductal shunting and if there is persistence of a ductal murmur on auscultation. PMID- 11399344 TI - The feasibility and safety of fluoroscopy-guided percutaneous intramyocardial gene injection in porcine heart. AB - BACKGROUND: Catheter-based transendocardial gene injection would be useful for the delivery of genes into the heart. We examined the feasibility and safety of percutaneous intramyocardial gene injections with fluoroscopic guidance alone. METHODS: We performed the procedure through an 8F arterial sheath inserted into the left carotid artery. In protocol 1, a mixture of India ink and normal saline was injected through a needle injection catheter in six pigs. We monitored blood pressure and ECG continuously during the procedure. Echocardiography, left ventriculography, and coronary angiography were performed. All pigs were sacrificed 2 days later and hearts were harvested. In protocol 2, a mixture of India ink and plasmid encoding CAT gene was injected in the same manner in eight pigs. Myocardial tissue was obtained 7 days after the procedure to assess gene expression. In protocol 3, four pigs were intentionally needle-perforated in the ventricular wall and were observed for 7 days. RESULTS: In protocol 1, there was no significant hemodynamic changes or serious arrhythmias during the procedure. Echocardiography and angiography revealed no evidence indicating pericardial effusion or wall motion abnormalities. Harvested hearts revealed one intramyocardial hematoma in a total of 36 injection sites. In protocol 2, the gene expression could be identified in 39 sites out of 48 injections after 7 days. In protocol 3, no animal showed signs indicating cardiac tamponade during the observation period. CONCLUSIONS: Our data suggest that fluoroscopy-guided percutaneous intramyocardial gene injection is a feasible and safe procedure, with no indication of associated significant hemodynamic changes, arrhythmias, or mortality. PMID- 11399345 TI - A descriptive framework for country-level analysis of health care financing arrangements. AB - Health financing policies are marked by confusion between policy tools and policy objectives, especially in low and middle income countries. This paper attempts to address this problem by providing a conceptual framework that is driven by the normative objective of enhancing the 'insurance function' (access to needed care without financial impoverishment) of health care systems. The framework is proposed as a tool for descriptive analysis of the key functions, policies, and interactions within an existing health care system, and equally as a tool to assist the identification and preliminary assessment of policy options. The aim is to help to clarify the policy levers that are available to enhance the insurance function for the population as efficiently as possible, given the 'starting point' of a country's existing institutional and organizational arrangements. Analysis of health care financing systems using this framework highlights the interactions of various policies and the need for a coherent package of coordinated reforms, rather than a focus on particular organizational forms of 'health insurance'. The content of each main health care system function (revenue collection, pooling of funds, purchasing of services, provision of services) and the market structure with which the implementation of each is organized are found to be particularly important, as are policies with respect to the benefit package and user fees. PMID- 11399346 TI - Are publicly-insured psychiatric outpatients in Japan satisfied? AB - Japan has a government financed outpatient program for people with mental disorders, called the 'publicly-insured' program. This study was performed to examine whether the target patient population used this publicly-insured program properly and to compare the degree of satisfaction of publicly-insured psychiatric outpatients with generally-insured psychiatric outpatients. The characteristics and satisfaction of 97 (43.9%) publicly-insured psychiatric outpatients and 124 (55.1%) generally-insured outpatients in Japan were studied. Psychiatrists rated sociodemographic and diagnostic information and patients were asked to complete the Japanese version of Client Satisfaction Questionnaire (CSQ 8J). The publicly-insured were longer-term and lower functioning patients and were significantly more dissatisfied with the services they received than the generally-insured patients. The publicly-insured program was successful in that patients with lower functioning (the primary target population of this program) were cared for and because they received treatment for longer periods of time. However, the program does not sufficiently satisfy the consumers of the services, despite its high costs. In this respect, this program needs to focus more on patients' points of view. More information on programs their enrollment procedures for patients may be helpful in educating consumers and citizens, clarifying expectations of services, and in influencing satisfaction. PMID- 11399347 TI - Cost-effectiveness study on influenza prevention in Hong Kong. AB - INTRODUCTION: Recent studies confirm that influenza vaccination confers health benefits and reduces direct and indirect costs associated with the illness. However, these studies did not examine the situation in southern China, a hypothetical influenza epicentre for the emergence of pandemic influenza viruses. METHODS: Surveillance data were collected in Hong Kong in 1993/94 and used economic model was used to estimate the medical and social costs associated with influenza-like illness (ILI) and to predict the cost-effectiveness of implementing an influenza prevention programme. FINDINGS: The estimated ILI incidence was 110/1000. It was highest in those between 1 and 25 years of age while the rate of hospitalization was highest in the elderly. Influenza occurred throughout the study period, which was a mild influenza year. The model predicted more than 660000 ILI cases in a non-epidemic year, in which influenza B virus predominated, with an average ILI-associated cost of HK$283/case (US$36) and vaccination-associated costs of HK$74 (US$9.50) per vaccinated individual. CONCLUSION: The medical, social and monetary costs of ILI in Hong Kong were not observed to be large when compared with those in more developed countries where there is a clearly defined influenza season and recognized disease burden. From the perspective of a susceptible individual, the vaccine was cost-effective but from the perspective of society it was not, even with the most cost-effective strategy of targeting the elderly. However, if the vaccine were effective in controlling newly emerging and highly virulent strains, targeted vaccination programmes might be highly cost-effective. PMID- 11399348 TI - What is 'best practice' in health care? State of the art and perspectives in improving the effectiveness and efficiency of the European health care systems. AB - A framework for the classification of information on maintaining or improving effectiveness and efficiency in health care systems is proposed. Activities, disciplines and methods that are available to identify, implement and monitor the available evidence in health care are called 'best practice'. We reviewed the literature in order to (1) establish a definition for 'best practice' in the health sector, (2) develop a framework to classify relevant information, and (3) synthesise the literature on activities, disciplines and methods pertinent to the concept. Health care, public health activities and health policy should be advised by the best available evidence. Currently, the concept can be broken down into three activities (Health Technology Assessment (HTA), Evidence-Based Medicine (EBM), Clinical Practice Guidelines (CPGs)) by which evidence is synthesised either as an evidence base (EBM and most HTA) or in the form of recommendations (CPGs and some HTA) for different decision purposes in health care. These activities gain input mainly through four disciplines: clinical research, clinical epidemiology, health economics and health services research. The different disciplines are related to each other in three 'domains': (a) input, (b) dissemination/implementation and (c) monitoring/outcome. These provide evidence on (a) the (potential) effects of health care interventions and policies; (b) on ways to implement them; and (c) on ways to monitor their actual outcome. None of these separate approaches and activities exclusively forms a successful and all-embracing strategy to ascertain 'best practice'. A collective approach in the management of information is expected to add value to individual efforts. Resources should be devoted to increase quality and quantity of both primary and secondary research as well as the establishment of networks to synthesise, disseminate, implement and monitor 'best practice'. PMID- 11399349 TI - Why is there an ERN/Ne on correct trials? Response representations, stimulus related components, and the theory of error-processing. AB - The ERN or Ne is a component of the event-related brain potential that occurs when human subjects make errors in reaction time tasks. It is observed in response-locked averages, time-locked to the execution of the incorrect response. Recent research has reported that this component is present on correct response trials, thereby challenging the idea that the component is specifically related to error-processing. In this paper, we argue that the ERN or Ne observed on correct trials can be attributed to one or both of two factors: either there is error-processing on correct trials, and/or the response-locked averages used to derive the ERN/Ne are contaminated by negative components evoked by the stimulus. For this reason, there is no reason to abandon theories that relate the ERN/Ne to error-processing. PMID- 11399350 TI - Error-negativity and positivity as they relate to other ERP indices of attentional control and stimulus processing. AB - We compared individual differences in the ERP associated with incorrect responses in a discrimination task with other ERP components associated with attentional control and stimulus discrimination (N2, P3, CNV). Trials with errors that are detected by the subject normally produce a negativity (N(E)) immediately following the response followed by a positivity (P(E)). The morphology of the N(E) and the P(E) is similar to that of the standard N2-P3 complex on correct discrimination trials. Our findings suggest that the P(E) is a P3 response to the internal detection of errors. The N(E), however, appears to be distinct from the N2. Finally, even though both the contingent negative variation (CNV) and the N(E) are associated with prefrontal cortex and the allocation of attention to response accuracy, the N(E) and CNV did not relate to one another. PMID- 11399351 TI - On the relationship of P3a and the Novelty-P3. AB - Deviant stimuli give rise to a late positive ERP component with latencies from 250 to 400 ms. Target deviants elicit a P300 with maximum amplitude over parieto central recording sites while the 'P300' elicited by deviant nontarget stimuli occurs somewhat earlier and shows a more frontally-oriented scalp distribution. Two varieties of frontal P300s have been described, elicited either by rare stimuli (target or nontarget) presented in a two-stimulus oddball task (P3a) or by infrequent, unrecognizable stimuli presented in the context of a three stimulus oddball task (Novelty-P3). The Novelty-P3 has been observed in a number of subsequent studies; the P3a has not been extensively studied and both its significance and existence have been called into question. The present report describes a replication of two prototypical studies with 'frontal' P3s observed in each context. Application of factor analysis to the two sets of ERP waveforms does not support a distinction between these two components. PMID- 11399352 TI - The role of perceived control in physiological reactivity: self-efficacy and incentive value as regulators of cardiovascular adjustment. AB - The aim of this study was to verify the influence of self-efficacy and incentive value on cardiovascular reactivity. Ninety-six subjects were randomly assigned to four experimental groups in each of which the self-efficacy (high or low) and incentive value (high or low) were modified in relation to a mental arithmetic task. Subjects were led to believe that failure in this behaviour would result in the appearance of an aversive stimulus. Heart rate, systolic and diastolic blood pressure, pulse pressure and skin temperature were all recorded throughout task performance, and at two given moments prior to the task, in order to establish baseline values. Subjects with high self-efficacy experienced a smaller increase in heart rate and systolic pressure, a greater increase in diastolic pressure, greater reduction in skin temperature and a reduction in pulse pressure during task performance. The effects were moderated by the incentive value in such a way that the effects of self-efficacy on cardiovascular reactivity were stronger when the consequences of task failure were perceived as highly relevant (high incentive value). PMID- 11399353 TI - The influence of serum vitamin B12 and folate status on cognitive functioning in very old age. AB - This study examined the relationship between low levels of serum vitamin B12 and folic acid (FA) and cognitive functioning in very old age. The four subsamples of non-demented persons aged 75-96 years - normal B12/normal FA; low B12/normal FA; normal B12/low FA; and low B12/low FA, were matched for age and education. A battery of cognitive tests was administered including Clock Tests, Block Design, Trail Making Tests (TMT), Digit Span, and tests of verbal fluency. Subjects with low levels of vitamins showed deficits in Block Design, TMT-B, Digit Span Backward, and letter fluency, but not in the remaining tests. In general, the effects of FA exceeded those of B12. This pattern of results was interpreted to mean that elderly persons with low vitamin levels have difficulty when fast and accurate processing of novel information is required, but are quite efficient in utilizing pre-existing knowledge structures. PMID- 11399354 TI - Seasonal variation in chemical constituents of Santolina rosmarinifolia L. ssp. rosmarinifolia. AB - The seasonal variation of the essential oil extracts from the aerial parts of a Santolina rosmarinifolia population has been studied. Oil yields increased in the months of March, April, May and June. Oil concentration showed significant correlations with both precipitation (positive) and temperature (negative). Essential oil main components were sabinene, beta-pinene, myrcene, beta phellandrene, limonene, 1,8-cineole, artemisia ketone, terpinen-4-ol, capillene, ar-curcumene and beta-eudesmol. Monoterpenes like beta-phellandrene, limonene and 1,8-cineole showed a significant negative correlation with temperature, while capillene presented a strong positive correlation with precipitation. The rest of the essential oil components did not show any noticeable trend. PMID- 11399355 TI - Mandibular gland chemistry of four Caribbean species of Camponotus (Hymenoptera: Formicidae). AB - The volatile components of whole-body extracts of males, females and workers were analyzed in four species of Neotropical ants in the formicine genus, Camponotus. The species, C. kaura, C. sexguttatus, C. ramulorum and C. planatus, represent three different subgenera. Volatile mandibular gland components were found only in male extracts in three of the species. In C. ramulorum, volatile components were found in male and female reproductives and workers. 3,4-Dihydro-3,5 dihydroxy-6-methylpyran-4-one and octanic acid were found in different sets of three of the species. Methyl 6-methyl salicylate was found in two species and the isocoumarin, mellein, was found in a third species. The significance of the mandibular gland secretion for formicid systematics is discussed. PMID- 11399356 TI - Understanding clonal diversity patterns through allozyme polymorphism in an endangered and geographically restricted Australian shrub, Zieria baeuerlenii and its implications for conservation. AB - An electrophoretic genetic analysis utilising starch gel electrophoresis was employed to assess clonality in endangered Zieria baeuerlenii populations distributed over an area of less than a kilometre square in Nowra (NSW). Eleven enzymes systems encoded by 19 loci (41 alleles) when assayed to estimate levels of genetic diversity within and among populations, revealed moderate levels of genetic diversity. Despite finding seven loci being fixed at a particular locus, 20 unique multilocus genotypes/clones restricted to a particular population were detected within a sample of 179 ramets collected from throughout the range of the species. The probability of finding any clone produced by sexual reproduction is <0.0025 and is further supported by the fact that reproduction appears to be exclusively by vegetative spread and there is virtually no pollen viability and seed production. The lack of seed set in this area may be due to additional factors inhibiting sexual reproduction. Overall, such genetic studies play a crucial role in devising conservation and management strategies for rare and endangered taxa. PMID- 11399357 TI - Genetic diversity and phylogenetic relatedness among six endemic Pterostylis species (Orchidaceae; series Grandiflorae) of Western Australia, as revealed by allozyme polymorphisms. AB - Starch gel electrophoresis was employed to survey the allozyme polymorphism and phylogenetic relationships among 35 populations covering six closely related Western Australian endemic Pterostylis species (series Grandiflorae); viz P. rogersii, P. aspera, P. angusta, P. hamiltonii, P. scabra and P. aff. alata. The aim of this study was to determine intraspecific and interspecific genetic diversity and species relationships based on allozyme analysis. The frequencies of 56 alleles at 12 enzyme systems coded by 15 loci were determined along with a mean intraspecific genetic identity value. Allozyme markers clearly discriminated populations belonging to different species. Nei's genetic distance/identity co efficient was used to measure the level of genetic differentiation among populations and species. Based on these values, a dendrogram was constructed which revealed that all the populations clustered into groups corresponding to the respective species. Gene diversity analysis among all the species revealed total genetic diversity H(t) of 0.23 with co-efficient of gene differentiation 10% (G(st)=0.10). Mean genetic variability (H(e)=0.136, P=40%) was also higher than for other outbreeding plant species. Mean genetic identity coefficient of populations of all species was 0.859 which increased to 0.877 upon exclusion of P. aff. alata, indicating a high degree of similarity among all species except P. aff. alata which segregated distinctively from the rest. Overall, the investigation provided independent support for the morphological segregation of these taxa. PMID- 11399358 TI - Flavonoids of Lavoisiera, Microlicia and Trembleya (Melastomataceae) and their taxonomic meaning. AB - Leaves of species of three closely related genera of Melastomataceae, Lavoisiera, Microlicia and Trembleya were analyzed for their flavonoid composition. A total of 116 compounds were obtained, comprising 69 flavonol and 47 flavone glycosides. The common occurrence of flavones, including 6-oxygenated derivatives, characterize Lavoisiera (which often yielded methoxylated flavonols and flavones). Flavonols predominate in species of Microlicia. The flavonoid patterns of Trembleya are rather confusing, some species being akin to Lavoisiera, others to Microlicia. UPGMA analysis using the flavonoid aglycones as characters and the samples analyzed as OTUs gives no complete resolution for the three genera, but provides clusters combining exclusively or preferentially species of either Lavoisiera and Microlicia. Species of Trembleya emerge from the phenogram interspersed among species of the other genera. The data suggest the recognition of Lavoisiera and Microlicia and the lumping of species of Trembleya with either of these genera. If greater flavone diversity is viewed as indicative of further evolutionary advancement, shrubby habits in Microliceae (Lavoisiera and Trembleya) should be regarded as derived from herbaceous ones (Microlicia). PMID- 11399359 TI - Chemical diversification trends in Astragalus caprinus (Leguminosae), based on the flavonoid pathway. AB - Flavonoid glycosides of Astragalus caprinus (Leguminosae) were investigated; more than 30 glycosides were found in leaf material, based on the aglycones kaempferol, quercetin and their methylated derivatives. Among them 14 compounds were found in significant amounts and showed a contrasting distribution. They could be ordered into three groups: polyglycosides, acylated polyglycosides and methylated polyglycosides. The distribution of these compounds was studied within a large collection of individual plants harvested in Tunisia; the results showed a relationship between metabolic trends and ecological diversification. PMID- 11399360 TI - Composition and chemical polymorphism of the essential oils from Piper lanceaefolium. AB - Essential oils obtained by hydrodistillation from leaves and spikes of Piper lanceaefolium H.B.K. of Costa Rica were analysed by GC-FID, GC-MS and 13C-NMR methods. Main constituents found in the oil from leaves were sesquiterpene hydrocarbons - especially beta-caryophyllene and germacrene D - and phenylpropanoids, of which elemicin and parsley apiol were the major ones. The volatile oil from spikes showed monoterpene hydrocarbons, namely alpha- and beta pinene, and the same phenylpropanoids as in the oil from leaves as the major constituents. Results obtained in the analysis by GC-FID and GC-MS of the essential oils from individual plants of different geographic origin were submitted to chemometric cluster analysis and principal component analysis, showing the presence of three different types of oils (i) parsley apiol/elemicin, (ii) elemicin/parsley apiol/dill apiol, and (iii) parsley apiol/dill apiol. PMID- 11399361 TI - Alkaloids from Crinum macowanii. PMID- 11399362 TI - Alkaloids from Crinum lugardiae. PMID- 11399363 TI - Pyrazole alkaloids from Elytraria acaulis. PMID- 11399364 TI - Coumarins from Chaptalia integerrima (Asteraceae). PMID- 11399365 TI - Sideroxylin and 8-demethylsideroxylin from Eucalyptus saligna (Myrtaceae). PMID- 11399366 TI - Isoprenylated flavonoids from Tephrosia tuitoensis. PMID- 11399367 TI - Investigation of the C(16:3)/C(18:3) fatty acid balance in leaf tissues of Biebersteinia orphanidis Boiss. (Biebersteiniaceae). PMID- 11399368 TI - Cardiac radiology. Is there any role for radiology of the heart? PMID- 11399369 TI - Dimeric versus monomeric nonionic contrast agents in visualization of coronary arteries. AB - A cubital intravenous iodine contrast agent enhancement is used to visualize coronary arteries using EBT. The quality of the coronary artery visualization however is limited by the nearly simultaneous approximation of CT values in coronary arteries and myocardial tissue. The objective of the study was to evaluate if "under real clinical circumstances" the lower iodine concentration and the dimeric based characteristic of iodixanol may effect the kinetic of the applied contrast agent and the visualization of coronary arteries studied noninvasively by EBT. A double-blind, randomized, parallel study was performed in 111 cardiac patients, using iodixanol 270 mg I/ml or iohexol 300 mg I/ml. The kinetics of contrast enhancement was studied in the flow mode measuring following parameters: mean arrival time and mean time to reach peak CT values in the pulmonary trunk, transit time from the pulmonary trunk to the aorta as well as mean and maximum CT values in the left ventricular chamber and in the myocardium with respect to the body mass index. The mean difference of CT values in the left ventricular chamber and the myocardium was calculated. The length of the visualized coronary arteries was assessed and the diagnostic quality of coronary artery visualization scored on a visual analogue scale. Although iodixanol was used with a lower iodine concentration than iohexol there was no significant statistical difference between both groups with respect to the diagnostic visualization and length assessment of the coronary arteries as well as in the mean difference of CT values in the left ventricular chamber and the myocardium. This means that the advantageous dimeric characteristics of iodixanol may be used to reduce the amount of applicated iodine in contrast agents without loss of diagnostic image quality and information. PMID- 11399370 TI - MRI of the cardiomyopathies. AB - We examined the potentialities of Magnetic resonance imaging (MRI) in the evaluation of the main cardiomyopathies: hypertrophic, dilated, restrictive and arrhythmogenic right ventricular. The hypertrophic cardiomyopathy is generally adequately investigated by echocardiography, that well defines the myocardial thickening and the obstruction of the left ventricular output. However, by echocardiography we still have difficulties in the evaluation of the apex of the left ventricle and the right ventricle involvement. MRI provides a complete evaluation of the heart with a clear evidence also of the echocardiographic dark zones by means of a clear evidence of the apex of the right ventricle. The dilated form is also well investigated by MRI that provides a clear evaluation of the volumes, mass and ejection fraction by means of the 3D analysis including conditions of the ventricular remodelling. Moreover, this technique helps in the differential diagnosis of acute myocarditis. In the acute phase of myocarditis (first 2 weeks), in fact, the myocardium produces high signal intensity on the T2 weighted sequences due to the presence of oedema. The third form of cardiomyopathy is the restrictive one, characterised by reduced diastolic filling and diastolic volume, normality of the systolic function and parietal thickness, interstitial fibrosis and enlargement of both atria. The mean potentiality of MRI is related to the differential diagnosis with constrictive pericarditis. Only in the former, the pericardium appears irregularly thickened with areas exceeding 4 mm of pericardial thickness. Finally, the right ventricular arrhythmogenic cardiomyopathy represents the main indication to MRI evaluation. With this imaging modality we are can obtain a clear morpho-functional evaluation of the right ventricle and distinguish the intramyocardial adipose substitution characterised by areas of high signal in the myocardium. PMID- 11399371 TI - Feasibility of electron beam tomography in diagnosis of congenital heart disease: comparison with echocardiography. AB - OBJECTIVE: To evaluate the feasibility of electron beam tomography (EBT) for morphological assessment of congenital heart disease. MATERIALS AND METHODS: Sixteen subjects were examined by EBT, transthoracic echocardiography, cardiac cine angiography and surgery. EBT scan was performed using single slice continuous volume mode to acquire high-resolution static image. Double dose contrast medium was injected by bolus technique after measuring scan delay time and calculation of proper injection rate. The results of EBT and echocardiography were compared based on the results of cardiac cine angiography and surgical findings. All anomalous components were evaluated in all subjects and grouped according to major cardiac structures. Statistical analysis was performed to compare two modalities' ability to evaluate the anomalies of major structures. RESULTS: EBT was more sensitive to evaluate the anomalies of systemic vessels, pulmonary vessels and small systemic arteries such as coronary artery. EBT was less sensitive to identify the abnormality of cardiac valves such as valvular regurgitation. There was no difference of sensitivity in evaluation of cardiac chambers and septa between the two modalities. CONCLUSION: EBT assisted by transthoracic echocardiography is excellent noninvasive modality to diagnose congenital heart disease. PMID- 11399372 TI - Magnetic resonance imaging in coronary heart disease. AB - Modern level of cardiac magnetic resonance imaging (MRI) development already allows its routine use (with proper indications) in coronary heart disease patients for studies of heart morphology and functions, performance of stress tests for evaluation of myocardial perfusion and contractile function. Coronary MRA and some other new MR techniques are close to its wide-scale clinical application. It has been shown that cardiac MRI is a valuable tool for detection of postinfarction scars, aneurysms, pseudoaneurysms, septal defects, mural thrombi and valvular regurgitations. Due to intrinsic advantages of the method it is of special value when these pathological conditions cannot be fully confirmed or excluded with echocardiography. MRI is recognized as the best imaging method for quantification of myocardial thickness, myocardial mass, systolic myocardial thickening, chamber volumes, ejection fraction and other parameters of global and regional systolic and diastolic function. MRI is used in studies of cardiac remodeling in postinfarction patients. The most attractive areas for cardiovascular applications of MRI are assessment of myocardial perfusion and non invasive coronary angiography. Substantial progress has been achieved in these directions. There are some other new developments in studies of coronary artery disease with MRI. High-resolution MR is used for imaging and quantification of atherosclerotic plaque composition in vivo. Intravascular MR devices suitable for performing imaging-guided balloon angioplasty are created. But before MRI will be widely accepted by the medical community as a important cardiovascular imaging modality several important problems have to be solved. Further technical advances are necessary for clinical implementation of all major diagnostic capabilities of cardiac MRI. The subjective obstacles for growth of clinical applications of cardiac MRI are lack of understanding of its possibilities and benefits both by clinicians and radiologists themselves. So proper training of specialists and promotion of this promising modality among the medical community are necessary. PMID- 11399373 TI - The use of gated perfusion scanning in the assessment of left ventricular volume. AB - A total of 34 patients underwent both gated SPECT perfusion imaging and echocardiography, with each test blinded to the other. Wall motion analysis, calculation of ejection fraction and ventricular volume were obtained by each technique. A close correlation was observed between the ejection fractions calculated by each method (r=0.79). There was a less close relationship between the left ventricular volumes calculated by each method (r=0.61) this is probably due to the errors which result from geometric assumptions for each technique. Wall motion analysis revealed a good level of agreement between each method with no significant discrepancies between the techniques. PMID- 11399374 TI - Traumatic false aneurysm of the subclavian artery treated by insertion of Memotherm stent. AB - Traumatic aneurysms of the left subclavian artery and transverse cervical artery, subsequent to penetrating gunshot wound were diagnosed by angiography in 35-year old patient. Subclavian artery aneurysm was treated by insertion of the Memotherm bare stent, whereas the false aneurysm of the transverse cervical artery was embolized with Gianturco's coils. The follow up examinations at 6 and 12 months showed good patency of subclavian artery. PMID- 11399375 TI - Cystic adventitial degeneration of the popliteal artery-the diagnostic value of duplex sonography. AB - Cystical adventitial degeneration of the popliteal artery is a disorder which is difficult to diagnose, due to the similarity of the symptoms of people presenting with peripheral arterial occlusive disease (PAOD) or popliteal entrapment syndrome. The only thing that differs from patients suffering from PAOD is the lack of typical risk factors for arteriosclerosis. Typical diagnostic procedures like conventional angiography or magnetic resonance Imaging angiography can be negative, too and therefore misleading. The only which is crucial in the diagnosis of cystic adventitial degeneration of the popliteal artery is to know the morphological background of this disorder, namely that it is a cyst of the adventitia of the artery which leads to a dynamic exercise-dependent flow inhibition. We present a 57-year old white male who had a week's history of intermittent claudication in his left calf. He was lacking of typical risk factors for arteriosclerosis and on first examination all pulses in both lower extremities were palpable and Doppler index on both legs was >1. Only duplexsonography revealed a cystic formation impressing the left popliteal artery in the hight of the rift in the popliteal joint. PMID- 11399376 TI - Combination radiotherapy for hepatocellular carcinoma with intraductal tumor thrombus: a case report. AB - We report the successful treatment of hepatocellular carcinoma (HCC) associated with an intraductal tumor thrombus in a 67-year-old male. Abdominal ultrasonography (US) and computed tomography (CT) revealed intrahepatic biliary dilatation in the left hepatic lobe and an intraductal tumor thrombus. The main tumor lesion was not clearly visualized on abdominal US, dynamic CT, and hepatic angiography. We biopsied the intraductal tumor thrombus under US guidance. Histologically the biopsy specimen was a poorly differentiated HCC We thus diagnosed HCC with intraductal tumor thrombus. The total serum bilirubin level gradually rose to 3.1 mg/dl. This tumor was inoperable because of severe hepatic dysfunction. We chose to treat the patient with radiotherapy aimed only at the intraductal tumor thrombus because the main tumor was unclear. A percutaneous transhepatic biliary drainage (PTBD) tube was inserted into the common bile duct beyond the tumor thrombus and the tube was dilated. Once total serum bilirubin had reached the normal range, a combination of external beam radiation therapy (EBRT) plus an intraluminal brachytherapy, 192Ir boost was administered. The intraductal tumor thrombus was found to have vanished and the PTBD tube was removed. After this treatment, transcatheter hepatic arterial embolization was performed at the point of tumor appearance. This patient had a relatively long survival, approximately 30 months, with no clinical evidence of recurrent disease and biliary drainage was not necessary. PMID- 11399377 TI - Amebic abscess of urachal remnants. AB - We report a rare case of amebic abscess of the urachus, mimicking an urachal neoplasm: no previous reports of amebic infection of the urachus were found in the literature. The challenges of the differential diagnosis between urachal abscess and carcinomas based both on clinical and radiological data are discussed. PMID- 11399378 TI - MR staging of clinical stage I and IIa cervical carcinoma: a reappraisal of efficacy and pitfalls. AB - The purpose of this study was to evaluate the diagnostic efficacy and pitfalls of magnetic resonance (MR) imaging in preoperative staging of cervical cancer. MR imaging was performed to determine the tumor staging for 31 patients with cervical carcinoma emphasizing tumor size, parametrial invasion, vaginal invasion and lymph node metastases. Tumor size was 3.23+/-1.75 cm (mean+/-standard deviation) at MR imaging compared with 2.79+/-1.76 cm at surgical-pathologic evaluation. The discrepancy between the tumor size determined by MR imaging and the measured surgical specimens was consistent in tumors larger than 1 cm. In assessing parametrial invasion, vaginal invasion and lymph node metastases, MR imaging had an accuracy of 96.7 and 87%. In determining stage of disease and differentiating operable (< or =stage IIA) from advanced disease (> or =stage IIB), MR imaging had an accuracy of 83.8 and 96.7%. Pitfalls leading to staging errors included difficulties in differentiating cancer foci from surrounding tissue edema and excluding vaginal invasion in the presence of large cervical cancer. In conclusion, MR imaging is accurate in the evaluation of parametrial invasion and useful in the differentiation of operable from advanced disease. The ability of MR imaging to exclude vaginal invasion in the presence of large cervical cancer and differentiate cancer foci from surrounding tissue edema is not as reliable. PMID- 11399379 TI - Second malignancies in children: the usual suspects? AB - The aim of this article is to provide an up to date review of second malignant neoplasms (SMN's) following treatment for childhood cancer, referring to their incidence, the role of genetic factors, and how the primary malignancy and treatment received influence the type, site and prognosis of SMN's. The role of genetic factors will be discussed as far as they impact upon a predisposition to later development of SMN's. The primary malignancies that have important associations with SMN's will then be discussed, in particular Hodgkin's disease, retinoblastoma and acute lymphoblastic leukaemia. The important second malignancies will be highlighted, including tumours of the CNS and thyroid, osteosarcoma, secondary acute myeloid leukaemia and melanoma. Emphasis will be put upon identifying which patients are most likely to suffer from these tumours. An important part of the article are case histories. These are provided in combination with illustrations as a useful adjunct to the text, with a particular emphasis on radiological features, diagnosis and screening. Finally, the important but different roles of causal agents, in particular chemotherapy and radiotherapy are highlighted. PMID- 11399380 TI - Effects of a pediatric emergency department febrile infant protocol on time to antibiotic therapy. AB - The purpose of this study was to examine the effect of an Emergency Department (ED) protocol to reduce time to antibiotic administration in the febrile infant less than 3 months of age with a rectal temperature > or =38.0 degrees C. We conducted a before-after study of a febrile infant quality improvement initiative in an urban pediatric ED with approximately 35,000 patient visits per year. Records of infants less than 3 months of age presenting with a rectal temperature > or =38.0 degrees C, who underwent a full septic work-up (blood, urine, and cerebrospinal fluid studies, and possibly chest radiography), were identified by using daily ED logs. This review was performed in the month before and then 12 months after institution of the "Septic Infant Work-up Sheet" and a set of interventions (Febrile Infant Protocol) designed to streamline care of the febrile infant and to reduce the time to antibiotic administration. Data were analyzed by using the Kaplan-Meier survival estimate and the log-rank test. Patient demographic characteristics and severity of illness were similar across months; however, ED process of care was significantly changed. Initial analysis revealed a median time to antibiotics of 142 min. Subsequent analysis after implementation of the Febrile Infant Protocol revealed a median time to antibiotics of 105 min. This represents an overall time reduction of 25% from time of presentation to antibiotic administration. In conclusion, a guideline based ED febrile infant protocol changed clinical practice and improved time to antibiotics. PMID- 11399381 TI - Performance and interpretation of focused right upper quadrant ultrasound by emergency physicians. AB - The objectives of this study were to determine the accuracy of Emergency Physicians (EP) performing focused right upper quadrant (RUQ) ultrasound, to quantify how sonographic experience affects accuracy for gallbladder pathology, and to establish the time needed to complete a focused RUQ ultrasound. A convenience sample of patients with suspected gallbladder disease received a focused RUQ ultrasound by an EP. Sonographic findings, number of previous RUQ ultrasounds performed, and time for examination completion were recorded. Each patient then had a formal RUQ ultrasound by a sonographer blinded to the focused RUQ ultrasound results. Focused RUQ and formal ultrasound findings were compared, with the exception of the sonographic Murphy sign, which was compared to pathology reports. One hundred nine patients were enrolled. Fifty-one had gallstones. Forty-nine were detected by EPs, yielding a sensitivity of 96% [95% confidence interval (CI).87-.99]. Of the 58 patients without gallstones, 51 were correctly diagnosed by EPs (specificity = 88%, 95% CI.77-.95). The sonographic Murphy sign was present during 54 emergency examinations, but in only 24 formal studies. When compared to pathology reports, the emergency sonographic Murphy sign had a sensitivity of 75% compared to the formal ultrasound sensitivity of 45% for acute cholecystitis. EPs were less accurate for other sonographic findings, and level of experience had little effect on sensitivity or specificity for detecting gallstones. Eighty-three percent of emergency studies were completed in less than 10 min. Gallstones are accurately detected by EPs in a timely fashion. Additionally, compared to the radiologist's interpretation, the EP-detected sonographic Murphy sign was more sensitive for diagnosing acute cholecystitis. PMID- 11399382 TI - Thrombotic thrombocytopenic purpura presenting as bilateral flank pain and hematuria: a case report. AB - Thrombotic thrombocytopenic purpura (TTP) is a rare disease whose incidence is now increasing. We present a case of a 37-year-old man who presented with bilateral flank pain and hematuria, subsequently diagnosed with TTP. Thrombotic thrombocytopenic purpura has classically been characterized by the pentad of fever, microangiopathic hemolytic anemia, neurologic symptoms, renal dysfunction, and thrombocytopenia. The pathogenesis of the disease has been a mystery until recently. We review the current literature regarding the pathophysiology and management of this disorder. Our discussion focuses on the importance of understanding this disease while considering the differential diagnosis of a patient presenting with anemia and thrombocytopenia because the common pitfall of rapidly administering platelets to a patient with TTP may lead to a disastrous outcome. PMID- 11399383 TI - The carcinoid syndrome: an unusual cause of valvular heart disease. AB - A 30-year-old woman presented to the Emergency Department with complaints of shortness of breath, orthopnea, and a severe reduction in exercise tolerance. The symptoms were the result of severe valvular heart disease that resulted from a bronchopulmonary carcinoid tumor. The carcinoid syndrome is a distinctive clinical syndrome seen in patients with carcinoid tumors. Cardiac valvular lesions are seen in the majority of patients with the carcinoid syndrome and represent the most clinically significant consequence of the carcinoid syndrome. This case report discusses carcinoid tumors, the carcinoid syndrome induced by these tumors, and the therapeutic options in the management of carcinoid tumors. PMID- 11399384 TI - Cimetidine-induced dystonic reaction. AB - A 39-year-old woman presented to the Emergency Department complaining of nausea and vomiting. The patient was given intravenous cimetidine for epigastric pain and subsequently developed a dystonic reaction. Administration of cimetidine, an H2 receptor antagonist, is an uncommon cause of dystonic reaction. We discuss the pathophysiology, diagnosis, and treatment. PMID- 11399385 TI - A case of withdrawal from the GHB precursors gamma-butyrolactone and 1,4 butanediol. AB - We describe a case of withdrawal from the gamma hydroxybutyric acid (GHB) precursors gamma butyrolactone and 1,4-butanediol. Symptoms included visual hallucinations, tachycardia, tremor, nystagmus, and diaphoresis. Administration of benzodiazepines and phenobarbital successfully treated the withdrawal symptoms. As predicted from the metabolism of gamma butyrolactone and 1,4 butanediol to GHB, the symptoms were nearly identical to those reported from GHB withdrawal. Because GHB is now illegal in the United States, individuals have begun abusing the legal and easier to acquire GHB precursors. More frequent cases of both abuse and withdrawal from these GHB precursors can be expected. PMID- 11399386 TI - The prehospital electrocardiogram. AB - Although widely used in Europe, the prehospital 12-lead electrocardiogram (EKG) has seen only limited use in this country. Reported benefits of the 12-lead EKG include shortening the door-to-needle time, accelerating the initiation of reperfusion therapy, and overall improving the prehospital and hospital management and outcome of patients with acute myocardial infarction. The field EKG also provides the basis for prehospital fibrinolysis. Concerns still exist, however, regarding the best means of providing real-time field interpretation of the prehospital EKG and the potential for inappropriate field time delay, triage, and treatment of patients. Moreover, questions remain about the overall clinical and cost benefit of expanding this resource universally. The following article reviews the role of prehospital EKG in caring for patients with acute coronary syndromes. PMID- 11399387 TI - Triage in the trauma bay with the focused abdominal sonography for trauma (FAST) examination. AB - Bedside ultrasonography has been applied to the evaluation of blunt trauma patients for over a decade. The Focused Abdominal Sonography for Trauma (FAST) examination has been used to successfully triage blunt trauma patients. Although not traditionally thought to be as useful in penetrating trauma patients, ultrasound can help determine the extent of injury especially of the heart. We present two cases of multiple-stabbing victims who arrived at our Level I trauma center at the same time, when our trauma system was particularly overburdened by multiple consecutive traumas. The FAST examination helped us to accurately determine which of the two patients required operative intervention first, despite that patient's appearance of relative hemodynamic stability in comparison to the other stabbing victim. PMID- 11399389 TI - Exercise-associated hyponatremia in marathon runners: a two-year experience. AB - This study was conducted to better define the pathophysiology, risk factors, and therapeutic approach to exercise-associated hyponatremia. Medical records from all participants in the 1998 Suzuki Rock 'N' Roll Marathon who presented to 14 Emergency Departments (EDs) were retrospectively reviewed to identify risk factors for the development of hyponatremia. Hyponatremic patients were compared to other runners with regard to race time and to other marathon participants seen in the ED with regard to gender, clinical signs of dehydration, and use of nonsteroidal anti-inflammatory drugs (NSAIDs). An original treatment algorithm incorporating the early use of hypertonic saline (HTS) was evaluated prospectively in our own ED for participants in the 1999 marathon to evaluate improvements in sodium correction rate and incidence of complications. A total of 26 patients from the 1998 and 1999 marathons were hyponatremic [serum sodium (SNa) < or =135 mEq/L] including 15 with severe hyponatremia (SNa < or = 125 mEq/L). Three developed seizures and required intubation and admission to an intensive care unit. Hyponatremic patients were more likely to be female, use NSAIDS, and have slower finishing times. Hyponatremic runners reported drinking "as much as possible" during and after the race and were less likely to have clinical signs of dehydration. An inverse relationship between initial SNa and time of presentation was observed, with late presentation predicting lower SNa values. The use of HTS in selected 1999 patients resulted in faster SNa correction times and fewer complications than observed for 1998 patients. It is concluded that the development of exercise-associated hyponatremia is associated with excessive fluid consumption during and after extreme athletic events. Additional risk factors include female gender, slower race times, and NSAID use. The use of HTS in selected patients seems to be safe and efficacious. PMID- 11399390 TI - Altered mentation and seizure. PMID- 11399391 TI - Xanthogranulomatous pyelonephritis. PMID- 11399392 TI - Pharyngeal foreign body. PMID- 11399393 TI - Scalp necrosis and visual loss due to giant cell arteritis. PMID- 11399394 TI - Chest X-ray in a young woman demonstrating achalasia cardia. PMID- 11399395 TI - Academic productivity in emergency medicine. PMID- 11399396 TI - Experiences after the earthquake. PMID- 11399397 TI - Communicating advance directives from long-term care facilities to emergency departments. AB - Many residents of long-term care (LTC) facilities are transferred to Emergency Departments without advance directives (AD). The goal of this study was to describe an ideal model for the transfer of AD from LTC facilities to Emergency Departments. Health care providers were asked to describe their ideal model for the completion and transfer of the ADs of LTC residents. A grounded theory methodology was used to identify significant themes. The model we present as a result of this analysis acknowledges the importance of simplifying and standardizing ADs, but focuses more attention on the process of completing and transferring the AD. A key feature of this model is an emphasis on the education of LTC residents and their relatives about ADs and advance-care planning. This education should involve a variety of resources used in creative ways; it should begin as soon as LTC placement is being considered, and the emphasis should be on providing information and discussing options rather than pressuring residents to make a decision. PMID- 11399398 TI - Performance. PMID- 11399400 TI - You're interested in emergency medicine...now what? A year-by-year guide. PMID- 11399401 TI - Nutrition and survival in patients with liver cirrhosis. AB - Although the effect of malnutrition on survival has been demonstrated by a number of studies, it is not clear whether malnutrition represents an independent risk factor in patients with liver disease. We studied 212 hospitalized patients with liver cirrhosis who were followed clinically for 2 y or until death. Body fat and muscle mass were evaluated by triceps skinfold thickness (TSF) and midarm muscle circumference (MAMC), respectively. Multivariate analysis according to Cox's model assessed the predictive power of nutritional parameters on survival. Thirty four percent of patients had severe malnutrition as determined by MAMC and/or TSF below the 5th percentile and 20% had moderate malnutrition (MAMC and/or TSF < 10th percentile). Twenty-six percent of patients were overnourished (MAMC and/or TSF > 75th percentile). Severely and moderately malnourished patients had lower survival rates than normal and overnourished patients. When analyzed with Cox's regression analysis, severe depletion of muscle mass and body fat were found to be independent predictors of survival. The inclusion of MAMC and TSF in the Child Pugh score, the prognostic score used most with liver disease, improved its prognostic accuracy. The prognostic power of MAMC was higher than that of TSF. These data demonstrate that malnutrition is an independent predictor of survival in patients with liver cirrhosis. The inclusion of anthropometric measures in the assessment of these patients might provide better prognostic information. PMID- 11399402 TI - Eating disorders: a role for dipeptidyl peptidase IV in nutritional control. AB - Dipeptidyl peptidase IV (DPP IV), a serine protease with broad tissue distribution and known activity in serum, has been postulated to modulate nutrition control by modification or inactivation of peptide hormones operating in the enteroinsular axis. We hypothesized that changes of DPP IV activity in serum are related to the nutrition status of patients with eating disorders. Serum DPP IV activity was measured in 52 patients (28 with anorexia nervosa and 24 with bulimia nervosa) in four consecutive weekly analyses. Simultaneously, the number of CD26 (DPP IV)-positive peripheral blood lymphocytes was counted. The same analyses were carried out in 28 healthy female volunteers. In week 1 and throughout the observation period, DPP IV activity in the sera of patients with anorexia nervosa and, to a lesser extent, those with bulimia nervosa was elevated in comparison to that of healthy controls (week 1: means = 92.8 U/L for anorexia nervosa patients and 89.3 U/L for bulimia-nervosa patients versus 74.7 U/L for healthy control subjects, P = 0.014; weeks 1-4: 91.8 U/L for anorexia-nervosa patients and 86.2 U/L for bulimia-nervosa patients versus 77.6 U/L for healthy controls, P < 0.001). We assume that the increase in DPP IV serum activity will increase the turnover of distinct peptide hormones with known effects on nutrition control and susceptibility to degradation by DPP IV. The potential impact of an increase in DPP IV activity in serum on satiety and nutrition control contributes to previously reported implications for immune function. PMID- 11399403 TI - Effects of malnutrition during early lactation on development and feeding behavior under the self-selection paradigm. AB - Selection of food can be affected by several factors, and with the method of self selection, qualitative changes in nutritional balance may be detected. The goal of the present study was to evaluate feeding preferences in weaning rats using three macronutrients (protein, carbohydrate, and fat), through a free-choice method, evaluating the alteration in their feeding patterns as compared with the previous nutritional status during the early lactation period. We analyzed the effects of protein restriction during lactation over the nitrogen balance after the weaning. The dams were assigned to one of two diet conditions (nourished or malnourished). At weaning, two pups from each litter were housed individually in metabolic cages, and they were maintained on self-selection under a free-choice paradigm and were provided with separate sources of macronutrients. The parameter for evaluating the nutritional effectiveness of the diets was nitrogen balance. We observed that protein intake tended to increase and consumption of carbohydrate and fat tended to decrease progressively during the 3 wk of experiment. In selecting their own food, growing rats and malnourished rats consumed a larger amount of protein than the other rats. Nourished rats selecting their diet had a larger nitrogen balance than nourished rats receiving a composed diet; no nitrogen balance difference was found between the self-selecting groups. Rats can choose an adaptive form when recovering from protein malnutrition. PMID- 11399404 TI - Permeability characteristics of polyamines across intestinal epithelium using the Caco-2 monolayer system: comparison between transepithelial flux and mitogen stimulated uptake into epithelial cells. AB - The polyamines putrescine, spermidine, and spermine are present in foods in high amounts, and are used for cell growth throughout the body. Surprisingly little is known about the mechanisms of polyamine absorption in the gut. To elucidate the mechanisms, transepithelial transport of polyamines was studied in human enterocytelike Caco-2 cells, grown on permeable filter supports. Transport of all three polyamines across Caco-2 cell monolayers was linear; intraepithelial accumulation of polyamines was higher in confluent than in differentiated Caco-2 cells, but still negligible in comparison with the overall transport across the monolayers. Epidermal growth factor (EGF) enhanced polyamine accumulation in Caco 2 cells four-fold, and basolateral uptake was higher than apical uptake if the cells were stimulated to grow. The amounts of polyamines taken up by the cells were nevertheless negligible in comparison with the net polyamine flux across the monolayers. Basolateral excretion of polyamines was in the picomolar range, whereas their transepithelial transport, occurring presumably by passive diffusion through the paracellular pathway, contributed hundreds of micromoles of polyamines to the basolateral chamber. We conclude that transepithelial transport of polyamines occurs by passive diffusion, and that it is not influenced when epithelial cells are stimulated to proliferate by a potent mitogen such as EGF. PMID- 11399405 TI - Differential effects of diets that provide different lipid sources on hepatic lipogenic activities in rats under ad libitum or restricted feeding. AB - This work was designed to study the effect of different lipid sources on hepatic lipogenic enzyme activity in rats fed ad libitum or energy-controlled diets. Male Wistar rats were fed diets containing 40% of energy as fat (olive oil, sunflower oil, palm oil, or beef tallow) for 4 wk. In experiment 1 rats had free access to food, and in experiment 2 rats were fed a controlled amount of food. In both experiments, rats fed the olive oil diets had higher activities of glucose-6 phosphate dehydrogenase, malic enzyme, fatty acid synthase, and acetyl-CoA carboxylase (P < 0.05) than rats fed the other fats. It is unlikely that this effect could be attributed to the stimulation by insulin or triiodothyronine because serum values did not differ among the groups. Enzymatic activities were positively and significantly correlated with liver triacylglycerol content, but not with serum triacylglycerol levels. No interaction between lipid source and feeding protocol was found. Oleic acid and components in olive oil other than fatty acids, such as phytosterols, may account for the effects of dietary fat on lipogenic enzyme activity. PMID- 11399406 TI - Modulation of epidermal growth factor-induced cell proliferation by an omega-3 fatty-acid-containing lipid emulsion on human pancreatic cancer cell line Mia Paca-2. PMID- 11399407 TI - Influence of glutamine-enriched parenteral nutrition on nitrogen balance and immunologic status in patients undergoing elective aortic aneurysm repair. PMID- 11399408 TI - Perioperative supplementation of EPA reduces immunosuppression induced by postoperative chemoradiation therapy in patients with esophageal cancer. PMID- 11399409 TI - Higher levels of inflammation in obese children. PMID- 11399410 TI - A good start in life: breast is best, but complementary foods should not be worse. PMID- 11399411 TI - Night-eating syndrome in obesity. PMID- 11399412 TI - Glutathione--nutritional and pharmacological viewpoints: part II. PMID- 11399413 TI - Clustered data. PMID- 11399414 TI - Sex differences in disease anorexia. AB - Sexually differentiated responses occur in molecular, cellular, physiologic, and organismic aspects of immune-system function in relation to acquired and innate immunities. These sex differences apparently include activational effects, which depend on gonadal hormone levels in adults, and lifelong effects, which arise directly from genetic differences or organizational effects of gonadal hormones early in development that lead to lifelong sex differences. Sex differences in immune function also can have great biological significance. Despite this, the mechanisms of these effects rarely have been analyzed extensively. This is especially true of anorexia during illness or disease. Therefore, this review briefly considers 1) the biological mechanisms of sex differences; 2) sex differences in immune function; 3) clinical and experimental data related to sex differences in four diseases or disease models that involve anorexia, Crohn's inflammatory-bowel disease, cancer, turpentine inflammation, and lipopolysaccharide bacteremia; and 4) sex differences in anorexia after interleukin-1 administration. PMID- 11399416 TI - Shortcomings of harm reduction: toward a morally invested drug reform strategy. AB - After a decade of steady diffusion in the drugs field, the harm reduction movement, posing pragmatic public health solutions based on empirical analysis, is still hindered by dissension and general confusion as to its underlying ideals. Despite having short-term political advantages, its 'value-neutral' style of discourse undercuts deeper moral foundations by attempts to forge the common ground in drug debates. Drawing on key statements in the literature and insights from interviews with leading Canadian drug policy observers, this commentary looks at rhetorical shortcomings that may act to encumber longer term harm reduction adoption and promotion. PMID- 11399417 TI - Supply and demand for alcohol in Australia: relationships between industry structures, regulation and the marketplace. AB - Aspects of alcohol supply and demand relationships are examined in relation to the two main beverage varieties in Australia, wine and beer. It is argued that this case study illustrates how the 'supply side' is able to create and protect demand for alcohol through both taking advantage of and influencing government regulation of the market for alcohol. In relation to low alcohol beer the impact on public health and safety has been extremely positive. In relation to the creation of cask wine in the late 1960s there have been demonstrably deleterious effects. Preferential taxation arrangements for the Australian wine industry have dramatically increased both exports and home consumption. One unintended consequence has been the creation of a major new market for cheap bulk wines that have had a devastating public health impact, particularly on Aboriginal communities, and also the invention of 'alco-pops'. Two-thirds of all table wine consumed in Australia now comes in a cask and 90% of this product is manufactured by three multi-national companies that wield enormous power and political influence to maintain the status quo. The Australian beer industry is well known internationally for its export of 'full strength' (around 5% by volume) beers. What is less well known is its commercial success in the development of low and mid-strength varieties for home consumption. In some States these now comprise 40% by volume of the beer market. This development can largely be attributed to State taxation arrangements, to drink-driving law enforcement, marketing strategies and to a decade of intense competition between several major brewers. This case study indicates how alcohol taxation policy can have major impacts on public health (both positive and negative) but that in a modern market economy it is difficult for governments to act in the public interest due to pressures from vested interest groups. PMID- 11399418 TI - The effectiveness of strategies such as health warning labels to reduce alcohol related harms - an Australian perspective. AB - The efficacy of health warning labels for products such as alcoholic beverages continue to be debated internationally and now in Australia as a means of mitigating the misuse of alcohol within community groups. This paper discusses evidence emanating primarily from the USA that has adopted a health warning label for both alcohol and tobacco, and from Australia that has adopted a health warning label for tobacco, on the effectiveness of such a strategy in changing consumer behaviour. The conclusion drawn is that such labelling is generally ineffective in changing consumer behaviour and hence such a strategy is inappropriate for reducing alcohol-related harms. The paper also discusses briefly what advertising and messages influence consumers, positively and negatively, and what specific strategies have been shown to better educate consumers and change their consumption from excessive to light to moderate as defined by the (Australian) National Health and Medical Research Council, which is the present premise of harm minimisation in Australia. PMID- 11399419 TI - Cracks in the conspiracy: the CIA and the cocaine trade in South Central Los Angeles. AB - This paper explores some of the issues surrounding the 1996 San Jose Mercury News "Dark Alliance" series; a story responsible for triggering heated debate on the idea that a US government agency conspired to use illicit drugs to undermine inner city African-American neighbourhoods during the 1980s. Beginning with an outline of Gary Webb's series and a discussion of the reaction it provoked, the paper looks at the issues behind the widespread sentiment that there is some form of federally controlled and racially inspired conspiracy. The paper examines the history of the CIA and its links with the illegal drug trade in an attempt to explain the prevalence of such a belief. Having identified some of the causes of what is often simplistically dismissed as baseless "black paranoia," the paper concludes that the idea that the CIA deliberately connived to target Afro Americans grossly oversimplifies the complexities of the inner city drug issue. Evidence suggests that during the cold war the CIA did indeed cultivate many dubious drug related alliances in the name of national security. It is argued here, however, that the notion of conspiracy based on skin colour merely detracts from the systemic failings of Washington's approach to drug control and ignores the Agency's indiscriminate pursuit of its anti-Communist agenda. PMID- 11399420 TI - 'Weddings, parties, anythingellipsis', a qualitative analysis of ecstasy use in Perth, Western Australia. AB - This study investigates the patterns of use, the meanings associated with use, the perception of risk and the strategies adopted to reduce these risks for a sample of ecstasy users in Perth, Western Australia. The sample was purposively chosen to represent the heterogeneous nature of ecstasy users in Perth. Data were collected from 31 ecstasy users and 157 h of participant observation in a variety of settings. The study found that users exhibited a reasonable degree of control over their consumption, incorporating a series of risk reduction strategies. Application of these strategies was inconsistent, with a large percentage of the sample indulging in occasional binges, spontaneous purchases, polydrug use and purchasing from unknown individuals in clubs/pubs. As users became more experienced, they tended to become less concerned about the risks associated with use and exhibited greater risk taking behaviour. Understanding the beliefs and events that influence the adoption (or non-adoption) of harm reduction strategies and the heterogeneous nature of ecstasy users is important for future health promotion interventions and policy. PMID- 11399421 TI - Neurobiology. PMID- 11399422 TI - Signalling mechanisms. Web alert. PMID- 11399424 TI - Trk receptors: mediators of neurotrophin action. AB - The four mammalian neurotrophins - NGF, BDNF, NT-3 and NT-4 - each bind and activate one or more of the Trk family of receptor tyrosine kinases. Through these receptors, neurotrophins activate many intracellular signaling pathways, including those controlled by Ras, the Cdc42/Rac/RhoG protein family, MAPK, PI3K and PLC-gamma, thereby affecting both development and function of the nervous system. During the past two years, several novel signaling pathways controlled by Trk receptors have been characterized, and it has become clear that membrane transport and sorting controls Trk-receptor-mediated signaling because key intermediates are localized to different membrane compartments. Three-dimensional structures of the Trk receptors, in one instance in association with a neurotrophin, have revealed the structural bases underlying specificity in neurotrophin signaling. PMID- 11399425 TI - The uniqueness of being a neurotrophin receptor. AB - Neurotrophins rely on Trk tyrosine kinase and p75 receptors for signal transduction. Recently, other roles for these receptors have been identified. Many questions have been raised about the mechanism by which these receptors mediate diverse cellular functions. Studies indicate a great deal of neurotrophin signaling specificity may stem from ligand-receptor selectivity and intracellular protein recruitment. PMID- 11399426 TI - Neuregulin and ErbB receptor signaling pathways in the nervous system. AB - The neuregulins are a complex family of factors that perform many functions during neural development. Recent experiments have shown that neuregulins promote neuronal migration and differentiation, and regulate the selective expression of neurotransmitter receptors in neurons and at the neuromuscular junction. They also regulate glial commitment, proliferation, survival and differentiation. At interneuronal synapses, neuregulin ErbB receptors associate with PDZ-domain proteins at postsynaptic densities where they can modulate synaptic plasticity. How this combinatorial network - comprising many neuregulin ligands that signal through distinct combinations of dimeric ErbB receptors - elicits its multitude of biological effects is beginning to be resolved. PMID- 11399427 TI - Transcription-dependent and -independent control of neuronal survival by the PI3K Akt signaling pathway. AB - The PI3K-Akt signaling pathway plays a critical role in mediating survival signals in a wide range of neuronal cell types. The recent identification of a number of substrates for the serine/threonine kinase Akt suggests that it blocks cell death by both impinging on the cytoplasmic cell death machinery and by regulating the expression of genes involved in cell death and survival. In addition, recent experiments suggest that Akt may also use metabolic pathways to regulate cell survival. PMID- 11399428 TI - Intracellular Ca(2+) pools in neuronal signalling. AB - Different intracellular pools participate in generating Ca(2+) signals in neuronal cells and in shaping their spatio-temporal patterns. They include the endoplasmic reticulum (endowed with different classes of Ca(2+) channels, with distinct functional properties and highly defined expression patterns in the brain), the Golgi apparatus, and the mitochondria. The release of Ca(2+) from intracellular pools plays an important role in controlling processes such as neurite outgrowth, synaptic plasticity, secretion and neurodegeneration. PMID- 11399429 TI - Mechanisms of Ca(2+)-dependent transcription. AB - Ca(2+) has a central role in coupling synaptic activity and transcriptional responses. Recent studies have focused on Ca(2+)-dependent nuclear mechanisms that bring to the nucleosomal level cascades of events initiated in the submembranous space at the synapse. In addition, a new Ca(2+)-dependent interaction between a calcium sensor and DNA has been shown to regulate transcription directly. PMID- 11399430 TI - How does calcium trigger neurotransmitter release? AB - Recent work has established that different geometric arrangements of calcium channels are found at different presynaptic terminals, leading to a wide spectrum of calcium signals for triggering neurotransmitter release. These calcium signals are apparently transduced by synaptotagmins - calcium-binding proteins found in synaptic vesicles. New biochemical results indicate that all synaptotagmins undergo calcium-dependent interactions with membrane lipids and a number of other presynaptic proteins, but which of these interactions is responsible for calcium triggered transmitter release remains unclear. PMID- 11399431 TI - NMDA receptor subunits: diversity, development and disease. AB - N-methyl-D-aspartate receptors (NMDARs) are present at many excitatory glutamate synapses in the central nervous system and display unique properties that depend on their subunit composition. Biophysical, pharmacological and molecular methods have been used to determine the key features conferred by the various NMDAR subunits, and have helped to establish which NMDAR subtypes are present at particular synapses. Recent studies are beginning to address the functional significance of NMDAR diversity under normal and pathological conditions. PMID- 11399432 TI - NMDA receptor regulation by Src kinase signalling in excitatory synaptic transmission and plasticity. AB - Regulation of postsynaptic glutamate receptors is one of the main mechanisms for altering synaptic efficacy in the central nervous system. Recent studies have given insight into the upregulation of the NMDA receptor by Src family tyrosine kinases, which bind to scaffolding proteins in the NMDA receptor complex. Src acts as a common step in signalling cascades that link G-protein-coupled receptors with protein kinase C via the intermediary cell-adhesion kinase beta. This signalling to NMDA receptors is required for long-term potentiation in the CA1 region of the hippocampus. PMID- 11399433 TI - NMDA receptor-mediated dendritic spikes and coincident signal amplification. AB - Dendrites of cortical neurons possess active conductances, which contribute to the nonlinear processing of synaptic information. Recently it has been shown that basal dendrites can generate highly localized spikes mediated by NMDA receptor channels. These spikes may serve as a powerful mechanism to detect and amplify synchronously activated spatially clustered excitatory synaptic inputs in individual dendritic segments, and may enable parallel processing in several integrative dendritic subunits. PMID- 11399434 TI - Ca(2+) signaling in dendritic spines. AB - Dendritic spines are cellular microcompartments that are isolated from their parent dendrites and neighboring spines. Recently, imaging studies of spine Ca(2+) dynamics have revealed that Ca(2+) can enter spines through voltage sensitive and ligand-activated channels, as well as through Ca(2+) release from intracellular stores. Relationships between spine Ca(2+) signals and induction of various forms of synaptic plasticity are beginning to be elucidated. Measurements of spine Ca(2+) concentration are also being used to probe the properties of single synapses and even individual calcium channels in their native environment. PMID- 11399435 TI - Coordinate regulation of metabotropic glutamate receptors. AB - Recent studies aimed at identifying the mechanisms that regulate the signaling of metabotropic glutamate receptors (mGluRs) have revealed that both protein kinase and protein phosphatase activity are important in directly modulating mGluR function. The inter-relationship between phosphorylation and dephosphorylation of mGluRs seems to be an important determinant in regulating mGluR function and the subsequent neuromodulatory events elicited by activation of mGluRs. PMID- 11399436 TI - Multiprotein complex signaling and the plasticity problem. AB - Synaptic transmission of distinct patterns of spikes, or 'neural code', leads to plastic changes in synapses and other parts of the neuron, as well as learning in animals. Recent findings indicate that specialized multiprotein structures associated with neurotransmitter receptors and cell-adhesion proteins function as molecular devices that both read the neural code and initiate long-term changes in synaptic structure and function. PMID- 11399437 TI - Allosteric mechanisms in normal and pathological nicotinic acetylcholine receptors. AB - Recent chemical and advanced structural studies on site-directed and naturally occurring pathological mutants of individual members of the multigene family of nicotinic acetylcholine receptors have yielded structure-function relationships supporting indirect 'allosteric' interactions between the acetylcholine-binding sites and the ion channel in signal transduction. PMID- 11399438 TI - Synaptic P2X receptors. AB - Over the past two years, ATP has clearly been shown to act as a co-transmitter with GABA, glycine and probably glutamate in the central nervous system. Our understanding of the ATP-gated P2X receptors is progressing rapidly, and the pharmacology, stoichiometry and subunit combinations of heteropolymeric P2X channels has been substantially elucidated. PMID- 11399439 TI - A neuron-glia signalling network in the active brain. AB - Glial cells are active partners of neurons in processing information and synaptic integration. They receive coded signals from synapses and elaborate modulatory responses. The active properties of glia, including long-range signalling and regulated transmitter release, are beginning to be elucidated. Recent insights suggest that the active brain should no longer be regarded as a circuitry of neuronal contacts, but as an integrated network of interactive neurons and glia. PMID- 11399440 TI - Use of administrative databases in occupational and environmental epidemiologic research. PMID- 11399441 TI - Assessment of vital status in Department of Veterans Affairs national databases. comparison with state death certificates. AB - PURPOSE: To determine the extent to which Department of Veterans Affairs (VA) database vital status information agrees with Washington state death certificates. METHODS: Using each data source, vital status was determined for 19,481 Washington state resident veterans hospitalized in Washington VA hospitals from 1994 to 1997, and for 33,602 Washington state resident veterans who were seen as outpatients during 1997. RESULTS: The agreement between VA and Washington state records was excellent for hospitalized veterans (kappa = 0.91, p < 0.0001). Three thousand one hundred-eight individuals (86.2% of all deaths) appeared in both files. Of those deaths missing in the VA files, 71% had no service-connected disability, VA pension, or other compensation. Among outpatients, agreement between the death files was very good (kappa = 0.82, p < 0.001). Three hundred seventy-two individuals (69.8% of all deaths) appeared in both files. Of those deaths missing in the VA files, 63% had no service-connected disability or VA pension or other compensation. CONCLUSIONS: The VA death files are a valid source of vital status information for veterans hospitalized in recent years. For veterans having exclusively outpatient visits, however, the VA files miss a substantial proportion of deaths. For these patients, alternative means of vital status ascertainment are warranted. PMID- 11399442 TI - Utilizing multiple vital status tracing services optimizes mortality follow-up in large cohort studies. AB - PURPOSE: To compare the three national-scale death identification services used in our two-stage vital status tracing protocol, Pension Benefit Information Company (PBI), Social Security Administration (SSA), and the Health Care Financing Administration (HCFA), with respect to death identification and confirmation rate, and relevant demographic variables. METHODS: Information on 31,223 subjects with unconfirmed vital status in an ongoing occupational cohort mortality study was simultaneously submitted to PBI, SSA, and HCFA to identify subjects deceased as of December 31, 1992. Subjects whose dates of death were between 1979 and 1992 were then sent to the National Death Index (NDI) to obtain death certificate numbers and supplemental states of death. RESULTS: PBI identified and confirmed the highest number deaths in this cohort. PBI and SSA identified a higher proportion of deaths for persons who died in earlier years and/or who died at a younger age, for both confirmed and unconfirmed deaths. HCFA identified fewer deaths overall and had a smaller proportion of unconfirmed deaths. These deaths occurred in later years among older subjects and had the highest proportion of females. NDI provided exact matches for 92-96% of deaths identified by each of the three services. CONCLUSIONS: PBI was the most comprehensive service, especially for identifying younger subjects and those with an earlier date of death, while HCFA may help to identify deceased female subjects. SSA data can be purchased and used for periodic updates or interactively to identify deaths among subjects with poor identifiers (such as incorrect or missing social security numbers or misspelled names). Because each service makes a valuable contribution to the identification of deceased cohort subjects, all three should be considered for optimal mortality follow-up. PMID- 11399443 TI - Population-based case-control study of occupational exposure to electromagnetic fields and breast cancer. AB - PURPOSE: This population-based case-control study examined occupational exposure to electromagnetic fields in relation to female breast cancer incidence among 843 breast cancer cases and 773 controls. METHODS: Exposure was classified based on work in the two longest-held jobs, and indices of cumulative exposure to magnetic fields based on a measurement survey. RESULTS: Female breast cancer was not associated with employment as an office or industrial worker. For the total study population, cumulative exposure over the entire career, and in the past 0-10 and 10-20 years generally showed odds ratios (ORs) close to the null. Moderately elevated risks were found for intermediate but not high levels of cumulative exposure accumulated 20 or more years ago (OR = 1.5; 95% CI = 1.1-2.0). Associations were stronger for premenopausal women (OR = 1.7; 95% CI = 1.1-2.7) in the past 10-20 years, and those with estrogen-receptor positive (ER+) breast tumors (OR = 2.06; 95% CI = 1.1-4.0). No consistent dose-response patterns were observed. CONCLUSIONS: These findings give little support to the hypothesis that electromagnetic fields cause cancer of the female breast. PMID- 11399444 TI - Serum dioxin and hepatic abnormalities in veterans of Operation Ranch Hand. AB - PURPOSE: We studied hepatic abnormalities and indices of hepatic function in relation to exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin (dioxin) in veterans of Operation Ranch Hand, the Air Force unit responsible for the aerial spraying of herbicides in Vietnam from 1962 to 1971. METHODS: The prevalence of ever having liver disease through March 1993, and level of alanine aminotransferase, aspartate aminotransferase, gamma-glutamyltransferase (GGT), lactic dehydrogenase (LDH), alkaline phosphatase, and total bilirubin were examined according to serum dioxin levels. RESULTS: We found an increased risk of "other liver disorders" among veterans with the highest dioxin levels [adjusted odds ratio (OR) = 1.6, 95% confidence interval (CI) 1.2 to 2.1], due primarily to increased transaminases or LDH (adjusted OR = 2.7, 95% CI 1.4 to 5.1) and to other nonspecific liver abnormalities (adjusted OR = 1.4, 95% CI 1.0 to 2.0). CONCLUSIONS: Whether the associations observed were causal is unclear from these data. PMID- 11399445 TI - Drinking pattern and mortality: the Italian Risk Factor and Life Expectancy pooling project. AB - PURPOSE: To analyze the relationship between an aspect of drinking pattern (i.e., drinking with or without meals) and risk of all-cause and specific-cause mortality. METHODS: The Risk Factors and Life Expectancy Study, is a pooling of a series of epidemiological studies conducted in Italy. Eight-thousand six-hundred and forty-seven men and 6521 women, age 30-59 at baseline, and free of cardiovascular disease, were followed for mortality from all causes, cardiovascular and noncardiovascular, during an average follow-up of 7 years. RESULTS: Drinkers of wine outside meals exhibited higher death rates from all causes, noncardiovascular diseases, and cancer, as compared to drinkers of wine with meals. This association was independent from the cardiovascular disease (CVD) risk factors measured at baseline and the amount of alcohol consumed and seemed to be stronger in women as compared to men. CONCLUSIONS: The present results indicate that drinking patterns may have important health implications, and attention should be given to this aspect of alcohol use and its relationship to health outcomes. The relationship between alcohol consumption and disease has been the focus of intensive scientific investigation (1-9). Most studies to date, however, have limitations. A major drawback is that limited information has been collected regarding the complex issue of alcohol consumption. In many studies, ascertainment of alcohol consumption frequently focused only on quantity of alcohol consumed without considering the many different components of alcohol consumption, particularly drinking pattern (10-12). It has been hypothesized, and preliminary data support the notion, that drinking pattern could have important influences on determining the health effects of alcohol (13,14). The present study examines the relationship between one aspect of drinking pattern (drinking wine outside meals) and mortality in a large cohort of men and women. PMID- 11399446 TI - A case-control study of physical activity and non-insulin dependent diabetes mellitus (NIDDM). the San Luis Valley Diabetes Study. AB - PURPOSE: The purpose of this study was to examine the association between physical activity and non-insulin dependent diabetes mellitus (NIDDM). METHODS: We conducted a population-based case-control study in Hispanic and non-Hispanic white men and women, ages 20-74. A total of 167 cases with NIDDM and 1100 controls with normal glucose tolerance were included. All subjects completed an oral glucose tolerance test. RESULTS: Persons with recently diagnosed NIDDM reported significantly lower levels of physical activity than control subjects. For total metabolic units, the odds ratio for subjects in the highest tertile compared to those in the lowest tertile was 0.60 (95% confidence interval (CI) = 0.37-0.98) after adjusting for age, sex, ethnicity, and family history of diabetes. The adjusted odds ratio for persons reporting high levels of vigorous activity (at least three times per week for 20 minutes) was also less than 1, but was not statistically significant (odds ratio (OR) = 0.73, 95% CI = 0.47-1.14). Similar adjusted odds ratios were observed for high versus low levels of self assessed work activity (OR = 0.50, 95% CI = 0.34-0.74) and leisure time physical activity (OR = 0.62, 95% CI = 0.44-0.90). Further adjustments for body mass index attenuated the strength of the association between physical activity and NIDDM. This is consistent with the hypothesis that obesity is one consequence of physical inactivity that puts individuals at increased risk for NIDDM. The association of physical activity and NIDDM was stronger in Hispanic than in non Hispanic white subjects, although this difference was not statistically significant. CONCLUSIONS: High levels of physical activity are associated with lower odds of NIDDM and this relationship may be stronger in Hispanic subjects. PMID- 11399447 TI - Prospective study of zinc intake and the risk of age-related macular degeneration. AB - PURPOSE: Zinc is found in high concentrations in the retina and is hypothesized to reduce the risk of age-related macular degeneration (AMD). Any long-term benefit associated with dietary zinc intake has not been evaluated. METHODS: We followed 66,572 women and 37,636 men who were > or = 50 years old and had no diagnosis of AMD or cancer. Zinc intake from food, multivitamins, and supplements was assessed with a semiquantitative food-frequency questionnaire at baseline (in 1984 for women and in 1986 for men) and repeated during follow-up (twice for women, once for men). RESULTS: During 10 years of follow-up for women and 8 years of follow-up for men, we confirmed 384 incident cases of AMD (195 cases of the early form and 189 cases of the late form) associated with a visual acuity loss of 20/30 or worse. After multivariate adjustment for potential risk factors, the pooled relative risk was 1.13 (95% confidence interval [CI], 0.82 to 1.57; p value, test for trend, 0.74) among participants in the highest quintile of total zinc intake (energy-adjusted median; 25.5 mg/day for women and 40.1 mg/day for men) compared with those in the lowest quintile (energy-adjusted median; 8.5 mg/day for women and 9.9 mg/day for men). The relative risk for highest compared with lowest quintile was 1.04 (95% CI, 0.59 to 1.83; p-value, test for trend, 0.54) for zinc intake from food. Subjects who took zinc supplements had a pooled multivariate relative risk of 1.04 (95% CI, 0.75 to 1.45). CONCLUSIONS: In these two large prospective studies, moderate zinc intake, either in food or in supplements, was not associated with a reduced risk of AMD. PMID- 11399448 TI - The effect of surfactant on birthweight-specific neonatal mortality rate, New York City. AB - PURPOSE: Surfactant has been shown to cause decreased neonatal mortality rate (NMR) in randomized studies of preterm infants. It is not clear whether the introduction of surfactant caused a decrease in neonatal mortality in a community. This study explores the hypothesis that the introduction of surfactant in 1990 to 1991 explains a decrease in neonatal mortality in New York City (NYC) among infants with birthweight of 500 to 1499 g. METHODS: For each of the 20 hospitals in New York City that began using surfactant in 1990 or 1991, we compared the NMR in the 2 years before the introduction of surfactant with the NMR in the 2 years after its introduction. Poisson regression models were fit to the death rates, adjusting for birthweight and other determinants of neonatal mortality. RESULTS: NMR in the 20 hospitals decreased by 13.7% (from 231.3 to 199.6 neonatal deaths per 1000 live births). This decrease remained significant after adjusting for birthweight and other risk factors. Infants with birthweight 1000 to 1249 grams benefited most from the introduction of surfactant; their NMR decreased by 19.6%. After adjusting for birthweight, those born before the introduction of surfactant were 1.18 times as likely to die in the first 27 days as those born after the introduction of surfactant (95% CI 1.04, 1.33). CONCLUSION: It appears that surfactant had a significant impact on NMR in NYC among very low birthweight babies. PMID- 11399449 TI - HIV Seroprevalence among newly incarcerated inmates in the Texas correctional system. AB - PURPOSE: The seroprevalence of HIV infection was examined among a sample of incoming inmates in the Texas Department of Criminal Justice prison system. Rates were compared across sociodemographic factors and three types of prison facilities: substance abuse felony punishment units (SAFPs), state jails, and prisons. METHODS: The study sample consisted of 4386 incoming inmates incarcerated for any duration, dating from November 1, 1998, to May 31, 1999. RESULTS: Among males, inmates entering state jails had a higher HIV infection rate (3.7%) than either inmates entering prisons (1.9%) or those entering SAFPs (0.5%). Among females, inmates entering prisons had a higher rate of infection (9.3%) than those entering state jails (2.5%) or SAFPs (4.5%). CONCLUSIONS: Although a number of blinded HIV seroprevalence studies have been conducted in U.S. prison systems, scarce information is currently available on HIV infection rates in alternative correctional facilities. The present study shows that HIV seropositivity varied substantially according to race, gender, and prison facility type. Given the shorter incarceration periods for inmates held in alternative facilities, understanding how infection rates vary according to type of incarceration facility holds particular public health relevance. PMID- 11399450 TI - Risk factors associated with moderate and serious injuries attributable to the 1994 Northridge Earthquake, Los Angeles, California. AB - PURPOSE: The purpose of this study was to use emergency department data to estimate levels of morbidity and risk factors due to earthquake-related mechanisms of injury subsequent to an urban night-time earthquake. METHODS: Data were abstracted from 4190 medical records for the month of January, 1994. Injuries attributable to the earthquake were identified through emergency department and medical records. These injuries were: (a) categorized by mechanism of injury; (b) assigned an injury severity score; and (c) linked to structural and geologic data. Proportional polytomous and dichotomous logistic regression were used to estimate risk of more severe injury associated with demographic characteristics, injury characteristics, structural characteristics, and geologic factors. RESULTS: More severe earthquake-related injuries (serious versus moderate and moderate versus minor) were statistically significantly associated with patient age (> or = 60 years old), upper extremities, falling, multi-family structures, pre-1960 housing, and the 50th percentile of Peak Ground Acceleration, after adjusting for all other available demographic, injury, structural, and geologic characteristics. CONCLUSIONS: The current recommendation of 'duck, cover, and hold' might not be optimal during a nighttime earthquake, particularly if individuals are in the padded environment of the bed. Actions such as reaching for or catching objects, bracing, or holding onto perceived stable objects may increase risk for more serious injury. Alternate responses include assuming a tucked position (as in airline crashes) or staying in bed for non-ambulating people. Structural damage and structure size were not associated with more serious injuries, but structure use and age were, leading the authors to suspect that unmeasured socioeconomic factors might impact risk factors. The importance of including population demographic characteristics in hazard modeling is emphasized. PMID- 11399451 TI - Proceedings of the Fourth International Symposium on Fish Endocrinology. Seattle, Washington, USA, 31 July-4 August 2000. PMID- 11399452 TI - Paradigms of growth in fish. AB - Most fish are indeterminate growers with white muscle making up the majority of the acquired bulk. Within the muscle, the myofibrillar fraction accounts for almost two-thirds of the protein synthetic activity, implying that it is accretion of myofibrillar proteins that makes the single most important contribution to fish growth. Fish muscle growth itself is not linear and occurs through a combination of hyperplasia and hypertrophy in post-juvenile stages. Superimposed on periodicity of growth in length and mass can be other phases governed by lunar, reproductive or circannual cycles. Data on fish growth are discussed in the framework of site-specific muscle abundance, metabolic and functional zonation of muscle, proliferation and differentiation of satellite cells and the contribution of myofibrillar proteins. Hormonal control of muscle growth is described against the backdrop of plasma availability of myogens (insulin, IGF-I, growth hormone), distribution and dynamics of their respective receptors, and their interactions. Important contributions of the 'supply side' are discussed with hormones regulating amino acid resorption from the intestine, intestinal growth, liver processing and amino acid uptake by the muscle. Data are also interpreted from metabolic angles, to explain lipolytic and nitrogen-sparing effects of growth hormones, and lipogenic effects of insulin and high protein diets. Finally, special attention is devoted to the multifaceted roles of arginine in fish growth, as precursor, intermediate and hormone secretagogue. PMID- 11399453 TI - Localization of somatostatin mRNAs in the brain and pancreas of rainbow trout (Oncorhynchus mykiss). AB - Rainbow trout possess three distinct mRNAs, each encoding a separate precursor: PPSS I, which contains a 14-amino acid sequence at its C-terminus (somatostatin 14) that is highly conserved among vertebrates, as well as two others, PPSS II' and PPSS II", both containing [Tyr(7), Gly(10)]-somatostatin-14 at their C terminus. In this study, we used RNA template-specific PCR and in situ hybridization to determine the distribution and cellular localization of PPSS mRNAs in the brain and Brockmann body of rainbow trout. PPSS I, PPSS II' and PPSS II" were expressed in the Brockmann body and pituitary; the expression of PPSS mRNAs in the brain was region specific. PPSS I mRNA was expressed in the Brockmann body predominantly by cells other than those that expressed PPSS IIs; however, there were several instances where PPSS I and PPSS IIs were co-expressed within the same cell. Of the PPSS II-expressing cells, many were observed to express both PPSS II' and PPSS II" mRNA; however, some cells expressed only PPSS II' mRNA, while other cells expressed only PPSS II" mRNA. In the brain, PPSS I mRNA was expressed in the optic tectum (OT) and in many hypothalamic nuclei, including the nucleus rotundus (NR), nucleus anterioris hypothalami (NAH), nucleus anterior tuberis (NAT), nucleus lateral tuberis (NLT), as well as in the pituitary (adenohypophysis). PPSS II" mRNA was present in the same regions as PPSS I mRNA; however, PPSS II' mRNA was present primarily in OT, NAT, NLT and adenohypohysis. These results indicate that PPSS mRNAs are expressed differently by different cells, suggesting that cell-specific mechanisms are involved with the control of PPSS expression and that particular biological responses may be associated with a specific SS isoform. PMID- 11399454 TI - Serum insulin-like growth factor binding proteins (IGFBPs) as markers for anabolic/catabolic condition in fishes. AB - In fishes as well as in all vertebrates in which it has been assessed, physiological shifts toward catabolism (e.g. such as during food deprivation) are consistently associated with elevations in the serum levels of at least one (often two in fishes) IGFBP in the < or =31-kDa size range. In mammals, 30-kDa IGFBP-1 is strongly up-regulated under catabolic circumstances, and it plays an important physiological role by sequestering IGF peptides to inhibit energy expensive growth until conditions are more favorable (e.g. with resumed feeding). Similarly in fishes, it has been found that when the < or =31-kDa IGFBPs are elevated in serum, somatic growth is inhibited, suggesting a similar growth inhibitory role of these proteins to that of mammalian IGFBP-1. Three different experimentally-induced catabolic states in fishes are compared in this paper: fasting; insulin-dependent diabetes mellitus (IDDM); and stress. A strong relationship between elevated serum cortisol concentrations and the presence of IGFBPs in each case is noted, and the utility of serum IGFBP measurement to serve as an effective indicator (marker) of catabolic condition in fishes is discussed. PMID- 11399455 TI - Insulin-like growth factor-I augments prolactin and inhibits growth hormone release through distinct as well as overlapping cellular signaling pathways. AB - We recently discovered a new role for insulin-like growth factor-I (IGF-I) as a specific and direct stimulator of prolactin (PRL) release in addition to its recognized function as an inhibitor of growth hormone (GH) release and synthesis. Little is known of the mechanisms that transduce the actions of IGF-I on PRL and GH release in vertebrates. The present study was undertaken to determine the cellular pathways that mediate the disparate actions of IGF-I on PRL and GH release in hybrid striped bass (Morone saxatilis X M. chrysops). When regulating cellular function, IGF-I may activate two primary pathways, phosphatidylinositol 3-kinase (PI 3-K) and mitogen-activated protein kinase (MAPK). The specific MAPK inhibitor, PD98059, blocked IGF-I-evoked PRL release as well as GH release inhibition over an 18-20-h incubation. LY294002, a specific PI 3-K inhibitor, overcame IGF-I's inhibition of GH release but was ineffective in blocking PRL release stimulated by IGF-I. These studies suggest IGF-I disparately alters PRL and GH by activating distinct as well as overlapping signaling pathways central for mediating actions of growth factors on secretory activity as well as cell proliferation. These results further support a role for IGF-I as a physiological regulator of PRL and GH. PMID- 11399456 TI - Glucose intolerance in teleost fish: fact or fiction? AB - Teleost fish are generally considered to be glucose intolerant. This mini-review examines some of the background and the possible mechanistic bases for this statement. Glucose intolerance is a clinical mammalian term meaning that a glucose load results in persistent hyperglycemia. Teleost fish show persistent hyperglycemia that is generally coincident with transient hyperinsulinemia. The fact that teleost generally have high plasma insulin compared with mammals implies insulin-deficiency is not a suitable explanation for this persistent hyperglycemia. Instead, peripheral utilization of glucose is probably the principle cause of hyperglycemia. Recent evidence for muscle insulin receptors, glucose transporters and hexokinase/glucokinase is reviewed and future experimental directions are suggested. If by altering peripheral glucose utilization fish could become more glucose tolerant, costs to the aquaculture industry may be substantially reduced. PMID- 11399457 TI - Alterations in the GnRH-LH system in relation to gonadal stage and Aroclor 1254 exposure in Atlantic croaker. AB - Exposure of Atlantic croaker (Micropogonias undulatus) to the polychlorinated biphenyl mixture (Aroclor 1254, PCB; 1 mg/kg body wt/day for 30 days) during the early-recrudescence phase of the gonadal cycle results in the impairment of LH secretion and gonadal growth. In order to determine whether impairment was due to disruption of the stimulatory GnRH neuroendocrine pathway, we compared various parameters of the GnRH-LH system in early recrudescing vs. spermiating (mature) fish. Seabream GnRH (GnRH) content in the preoptic anterior hypothalamic area (POAH) and pituitary, pituitary GnRH receptor concentrations, and basal and GnRH analog (GnRHa)-induced LH secretion were significantly higher in gonadally mature croaker compared to early-recrudescing fish. In a subsequent experiment, the effects of PCB on the same neuroendocrine indices were investigated during the gonadal recrudescence phase of croaker. PCB exposure during the period of testicular maturation prevented the natural increase in GnRH content in the POAH but not in the pituitary. This finding suggests that PCB may impair GnRH synthesis in the POAH. The number of pituitary GnRH receptors also remained significantly lower in the PCB-exposed group, which was likely due to an impairment of GnRH release. The GnRH content in the POAH, number of pituitary GnRH receptors, and LH secretion in the PCB-exposed group were comparable to those in early-recrudescing fish, suggesting an impairment of normal maturation of the GnRH-LH system during the gonadal recrudescence phase. This impairment may be due to a direct action of PCB on GnRH neurons and/or indirectly via interference with other neurotransmitter pathways that modulate GnRH function. PMID- 11399458 TI - Estrogen and xenoestrogens upregulate the brain aromatase isoform (P450aromB) and perturb markers of early development in zebrafish (Danio rerio). AB - Estrogen synthesized in the brain itself by the action of cytochrome P450 aromatase (P450arom) is known to have permanent organizing effects on the developing CNS. In fish, estrogen upregulates the predominant brain isoform (P450aromB), implying that xenoestrogens (XE) could act as neurodevelopmental toxicants by altering P450aromB. To test this hypothesis, zebrafish embryos were exposed to 17beta-estradiol (E(2)), diethylstilbestrol (DES, a potent agonist), and bisphenol A (BPA, a weak agonist). RT-PCR/Southern transfer analysis showed that E(2) (0.01-10 microM) upregulated P450aromB in a dose-response manner. The effect of DES (0.01 microM) was similar to 1 microM E(2) (three- to four-fold higher than control), but BPA was less effective ( 40 mm) hair were detected with CE-PTG and with histology. CE-PTG was also able to detect the severely miniaturised hair fiber (down to 8 mm diameter) and was comparable to scalp biopsy analysis. The latter could identify hair fibres, which did not reach the scalp surface, a measure that is considered as not clinically significant. All growth stages - anagen, catagen and telogen - as well as the empty follicle stage could clearly be observed with CE-PTG. Staging of the more severely affected hair follicles was not always possible neither with CE-PTG nor histology - even with serial sectioning. The finding of such technological advantages makes the CE-PTG a first choice method for detailed analysis of hair cycling in androgenetic alopecia - a scalp disorder characterised by extreme hair follicle miniaturisation, decreased hair pigmentation and hair thinning. PMID- 11399540 TI - Advances in the treatment of male androgenetic alopecia: a brief review of finasteride studies. AB - Finasteride is a type 2 5a-reductase inhibitor and therefore mimics the biochemical profile of inherited type 2 5a-reductase deficiency in men. It was developed to grow hair in androgenetic alopecia and shrink benign prostatic hyperplasia. Various clinical trials of finasteride have confirmed its beneficial effects in androgenetic alopecia in males, but not in females. It can produce visible hair growth in up to 66% of men with mild to moderate alopecia, but importantly can stop hair loss in 91% of patients. In long-term finasteride studies, placebo patients were characterized by significant and progressive hair loss. It can be concluded that finasteride prevents further hair loss by actually continuing to grow enough hair to preserve scalp coverage. This is confirmed by the loss of hair following withdrawal of finasteride in such cases. The proven preservative effect of finasteride, in addition to its restorative effect, is a strong indication for prescribing it in early cases of androgenetic alopecia before much hair has been lost. PMID- 11399541 TI - Costimulatory molecules and their ligands as therapeutic targets in autoimmune disease. AB - The initiation and progression of autoimmune diseases are complex processes that depend on a selective breakdown of tolerance and additional factors like genetic susceptibility and environmental factors, e.g. drugs, infections, toxins and UV light. The causative failure of the immune system to tolerate self can be initiated by molecular mimicry or polyclonal activation with the consequence of a breakdown of anergy or a failure of activation induced cell death. It presents itself in multiple forms including a disturbed balance between TH1 and TH2, alterations in the cytokine milieu and undue modulation of the expression of costimulatory molecules. Although all these features may vary between patients and within the individual patient depending on the state of disease, recent years have provided convincing evidence that, in particular, disease progression is markedly influenced by the expression profile of costimulatory molecules. Since in many forms of autoimmune diseases the causative self-antigen(s) are unknown, therapy largely depends on anti-inflammatory agents or in severe cases on a general immunosuppression. Increasing knowledge of the functional activities of costimulatory molecules in autoimmune disease now provides a new and promising therapeutic modality, which in a more selective way interferes with the pathological activities of immune cells. Here I discuss evidence for the involvement of costimulatory molecules, particularly of CD44 variant isoforms, in autoimmune diseases and their possible use as a therapeutic target. Due to the regulated and restricted expression of these molecules, treatment should not be burdened by severe side effects. PMID- 11399542 TI - Treatment of immune-mediated skin diseases: future perspectives. AB - In recent years there has been an enormous increase in the understanding of the pathogenesis of immune mediated skin diseases which has led to the development and introduction of new therapeutic regimes. Accordingly, non-specific immunomodulating drugs such as cyclosporin A (CyA), rapamycin, leflunomide, mycophenolat mofetil, tacrolimus, pimecrolimus (ASM981) a.o. proved to be beneficial for a variety of skin diseases. Immunomodulators such as tacrolimus, ASM981 and imiquimod also have been developed for topical application. Specific immunomodulating strategies involve humanized antibodies directed against cytokines or cell surface molecules, receptor antagonists, fusion proteins targeting cytokines or receptors and transcription factor inhibitors. Another interesting approach is to target the appropriate T cell-receptor on autoreactive T cells. Transfection with cytokine genes may represent a useful approach to generate immune deviation and, thereby, treat immune mediated diseases. The value of different vaccination strategies are currently investigated. The first promising results have been obtained by targeting the function of antigen presenting cells such as dendritic cells (DC). Although, the rapid development of research in immune-mediated diseases has led to the development of several new and more effective therapeutic strategies, in most cases cure is still not possible until the genetics of these diseases are revealed which ultimately may result in gene therapy. PMID- 11399543 TI - The molecular basis of hair growth. AB - For a long time, hair follicle development could be studied on the morphological level only. Now that molecular genetics is coming of age, we are beginning to understand the molecular basis of hair follicle development. The study of inherited hair disorders and basic research have both contributed to our insights and exciting developments can be expected in the near future. Here, we present a compact overview of the essential players in hair development and propose a simple model of the genetic interactions in the hair follicle. PMID- 11399544 TI - The hair follicle as a target for gene therapy. AB - The hair follicle possesses progenitor cells for continued hair follicle cycling and for epidermal keratinocytes, melanocytes and Langerhans cells. These different cell types can be targeted by topical gene delivery to mouse skin. Using a combination of liposomes and DNA, we demonstrated the feasibility of targeting hair follicle cells in human scalp xenografts as well. We defined liposome composition and stage of the hair cycle as important parameters influencing transfection of human hair follicles. Transfection occurred only during anagen onset. Considerations and obstacles for using gene therapy to treat alopecias and skin disease are discussed. A theoretical framework for future gene therapy treatments for cutaneous and systemic disorders is presented. PMID- 11399545 TI - Morphology of hair in normal and mutant laboratory mice. AB - Inbred laboratory mice are mammals and therefore are haired. Mice develop many of the same diseases as humans and have become the premier in vivo model for studying biology, pathology, genetics, and molecular mechanisms. Mice are useful tools to study hair biology. Examples of characterized mutant mice with abnormal pilosebaceous unit phenotypes are presented to illustrate the value of these animals as models to help understand human diseases of the skin and hair. PMID- 11399546 TI - TrichoScan: combining epiluminescence microscopy with digital image analysis for the measurement of hair growth in vivo. AB - Hair loss or hair thinning is a common complaint in clinical dermatology, and patients seeking advice for hair loss are not necessarily bald. Also the effects of treatment attempts are hard to measure. Consequently, there is a need for a sensitive tool to monitor hair loss and treatment response. Such a method must be able to analyze the biological parameters of hair growth, which are: 1: hair density (n/cm2), 2: hair diameter (mm), 3: hair growth rate (mm/day) and 4: anagen/telogen ratio. Here we present the TrichoScan as a method which combines epiluminescence microscopy (ELM) with automatic digital image analysis for the measurement of human, and potentially animal hair, in situ. The TrichoScan is able to analyze all biological parameters of hair growth with a so-called intraclass correlation of approx. ninety-one percent with the same TrichoScan operator and an intraclass correlation of approx. ninety-seven percent for different TrichoScan operators. The application of the technique is demonstrated by comparison of the hair parameters in individuals without apparent hair loss with men with untreated AGA and men after treatment with finasteride (1 mg/day), where we were able to detect a significant increase in hair counts and cumulative hair thickness 3 and 6 months after treatment. The advantage of the TrichoScan is that it can be used for clinical studies to compare placebo versus treatment or to compare different capacities of different hair growth promoting substances, it can be used for studying AGA or other forms of diffuse hair loss, and it can be adopted to study the effect of drugs or laser treatment on hypertrichosis or hirsutism. PMID- 11399547 TI - Trichoteiromania. AB - We describe a 61-year-old patient who compulsively rubbed her hair and her scalp because of a psychiatric disorder. Permanent rubbing resulted in fracturing of the hair shafts, leaving 2 cm long hairs and areas with stubs of 1mm length, giving the impression of bald spots. The distal ends of affected hair shafts were split, giving the impression of white tips. Light microscopy of the hair shafts showed split, brush-like ends of otherwise normal hair. We suggest the name trichoteiromania, which means "compulsive rubbing of hair", as a new term to describe hair loss in such cases. PMID- 11399548 TI - Hermansky-Pudlak syndrome. AB - A 55-year-old man had oculocutaneous albinism and a history of frequent bruising following minimal trauma. The simultaneous occurrence of these features was first described by Hermansky and Pudlak in 1959. The Hermansky-Pudlak syndrome follows an autosomal recessive trait and is most frequently found in Puerto Rico and in the Swiss alps. It consists of the triad phenotype of hypopigmentation, prolonged bleeding time due to platelet storage pool deficiency and accumulation of ceroid pigment in lysosomal organelles. Other serious features are pulmonary fibrosis and granulomatous colitis. The disorder is caused by mutations in the HPS1 gene on chromosome 10q23. The HPS1 gene product is involved in the trafficking of melanosomes, platelet dense bodies, and lysosomes. PMID- 11399549 TI - Malignancy: Subsequential Alterations of Telomeric DNA Length Correlate with Cytogenetic Response in Chronic Myeloid Leukemia Treated with Interferon alpha. AB - Telomeres consist of simple tandem hexametric (TTAGGG) repeats and progressively shorten with cell replication. To determine a relationship between telomeric erosion and response to treatment, we measured telomere length following treatment in patients with chronic myeloid leukemia (CML) in the chronic phase. We used 70 samples of bone marrow mononuclear cells obtained from 26 patients with CML in the chronic phase subsequently. Telomere length was determined by a Southern hybridization of HinfI-digested DNA using a (TTAGGG)(4) probe, and the terminal restriction fragment (TRF) length was measured. Telomerase activity was also measured in 14 CML patients at the time of diagnosis using a telomeric repeat amplification protocol (TRAP) assay. Of the 26 patients with CML at the time of diagnosis, 14 had normal TRF lengths and the remaining 12 had shortened TRFs compared to those of age-matched normal individuals. In a group of CML patients treated with interferon alpha (IFNalpha), 80% of those who showed normal TRFs obtained cytogenetic responses. Approximately 50% of patients with shortened TRFs and treated with IFNalpha showed normalization of TRFs after IFNalpha treatment and all of them were cytogenetic responders. None of the CML patients with shortened TRFs before and after IFNalpha treatment achieved major cytogenetic response and they had high levels of telomerase activity. In the group of CML patients treated with hydroxyurea alone, although some patients showed normalization of TRF lengths after treatment, none of them showed major cytogenetic response. Telomere length before treatment may be related to CML disease severity. Cytogenetic response could be expected in CML patients with normal TRF lengths and treated with IFNalpha. Thus, measurement of telomere length after treatment might provide important information in managing CML patients. PMID- 11399550 TI - Malignancy: Human T-Cell Lymphotropic Virus Type I and Adult T-Cell Leukaemia/Lymphoma. AB - Adult T-cell leukaemia/lymphoma (ATLL) was first identified in Japan in 1977 [1,2]. The causative agent, the human T-lymphotropic virus type I (HTLV-I), was isolated 3 years later by Gallo's group from a patient initially diagnosed as having mycosis fungoides but subsequently reclassified as a case of ATLL [3]. Since this time, much has been discovered about the molecular pathogenesis of the disease. Despite this, treatment of ATLL remains disappointing and the prognosis of acute and lymphoma types poor. In the United Kingdom, cases of ATLL are mainly restricted to people of Afro-Caribbean descent but the disease is of general importance because ATLL has also been reported in non-endemic areas and may possibly spread into other populations via blood transfusion as blood donors in the UK are currently not screened for HTLV-I. PMID- 11399551 TI - The Hematopoietic Microenvironment: Phylogeny and Ontogeny of the Hematopoietic Microenvironment. AB - Although there is no formalized area of study called phylogeny or ontogeny of the hematopoietic microenvironment, new models and molecular tools are now available for such studies. The concept of a hematopoietic microenvironment has developed from the need to answer basic questions about migration, control of proliferation and differentiation of lymphohematopoietic cells; e.g. how are cells with the same genes induced to express different sets of these genes which lead to differentiation. These questions were first approached when cells could only be identified morphologically. The ontogeny of hematopoiesis was traced from the blood islands of the embryonic yolk sac, to the fetal liver, spleen, and bone marrow. Cells with reticular morphology were associated with areas of hematopoiesis and, in the embryo, they were thought to give rise to both hematopoietic and supportive cells. In the 1960's the classic work of McCulloch, Till and Siminovitch led the study of hematopoietic precursors which have no distinctive morphological identity and are too infrequent to study microscopically. These cells were identified by their functions; e.g. colony formation in culture in the presence of certain factors, production of spleen colonies or rescue of lethally irradiated mice. Cells with these functions were also found sequentially in the yolk sac blood islands, in the aorta/mesonephros, fetal liver, spleen, and bone marrow during development. The question remained, what regulates the proliferation and differentiation of these cells and why do they home to different sites in different stages of development? Among the laboratories studying spleen colonies, a controversy arose as to whether differentiation decisions were stochastic or induced by extra cellular factors. Dexter and Greenberger developed the long-term bone marrow culture system which has aided in studying the roles of factors such as cell-cell contact and extracellular matrix in hematopoietic differentiation. The molecular identification of ligand/receptor pairs such as ckit and KL as well as transactivating factors that control whole sets of lineage related genes such as the GATAs and Ikaros, may lead to clarification of the stochastic versus induced differentiation issue. Chimeric bird and frog embryos and analysis of mutations effecting hematopoiesis in frogs and zebrafish have helped to trace the earliest hematopoietic development in the embryo and to determine what influences it. The identification of genes that alter development of hematopoiesis opens the possibility of comparing microenvironmental control mechanisms in various present day organisms and relating these to evolutionary events. Many basic questions relevant to the interaction between hematopoietic cells and their microenvironment can be addressed by studying "simple" organisms in which the answers may be more easily determined than in mice or humans. Examples of possibly useful organisms, range from the teliosts such as zebrafish to algae such as Volvox, a two cell organism, to Dictyostelium which change from 1 to many cell types and in the process, migrate, adhere and differentiate. PMID- 11399552 TI - The Hematopoietic Microenvironment: Heterogeneity of Bone Marrow Stromal Cells: Growth Properties and Cytokine Production by CD34+ and CD34- Stromal Sub-Sets. PMID- 11399553 TI - Molecular Medicine and Molecular Pathology for Haematologists: I. Gene Speak made Easy: An Overview of Genes and Gene Expression. AB - Molecular Medicine and Molecular Pathology are integral parts of Haematology as we enter the new millennium. Their origins can be linked to fundamental developments in the basic sciences, particularly genetics, chemistry and biochemistry. The structure of DNA and the genetic code that it encrypts are the critical starting points to our understanding of these new disciplines. The genetic alphabet is a simple one, consisting of just 4 letters, buts its influence is crucial to human development and differentiation. The concept of a gene is not a new one but the Human Genome Project (a joint world-wide effort to characterise our entire genetic make-up) is providing an invaluable understanding of how genes function in normal cellular processes and pinpointing how disruption of these processes can lead to disease. Transcription and translation are the key events by which our genotype is converted to our phenotype (via a messenger RNA intermediate), producing the myriad proteins and enzymes which populate the cellular factory of our body. Unlike the bacterial or prokaryotic genome, the human genome contains a large amount of non coding DNA (less than 1% of our genome codes for proteins), and our genes are interrupted, with the coding regions or exons separated by non coding introns. Precise removal of the intronic material after transcription (though a process called splicing) is critical for efficient translation to occur. Incorrect splicing can lead to the generation of mutant proteins, which can have a dilaterious effect on the phenotype of the individual. Thus the 100,000-200,000 genes which are present in each cell in our body have a defined control mechanism permitting efficient and appropriate expression of proteins and enzymes and yet a single base change in just one of those genes can lead to diseases such as haemophilia or fanconis anaemia. PMID- 11399554 TI - Erythropoiesis: Comparison of Cytotoxic Aldehyde Generation in Beta-Thalassemia Patients Chelated with Deferoxamine or Deferiprone (L1) Versus NO Chelation. AB - The mechanism of iron-induced organ failure in iron overload disorders is not known, but it is conjectured that excess iron-catalyzed free radical generation contributes to organ damage. We hypothesized that free radical generation, quantified by the presence of 20 separate cytotoxic aldehydes in plasma, would be significantly increased in non-chelated beta-thalassemia major patients, in comparison to those chelated with either deferiprone (L1) or deferoxamine (desferal). We also report on red cell glutathione peroxidase activity in these patient groups, an enzyme involved in averting the damaging effects of free radicals. Ten patients were chelated with nightly subcutaneous infusions of desferal and 10 received the experimental oral chelator L1. Body iron burden was assessed by serum ferritin and hepatic iron concentrations. In comparison to non chelated controls, significant decreases of 62% and 64% in total cytotoxic aldehyde concentrations were observed in patients chelated with desferal and L1, respectively (p < 0.001). Significantly lower red cell glutathione peroxidase activity was also observed in non-chelated controls, in comparison to those chelated with either desferal or L1 (p < 0.001). This is the first report on the concentrations of cytotoxic aldehydes in non-chelated beta-thalassemia major patients, and the first to report on the effects of L1 against cytotoxic aldehyde formation in plasma of patients with iron-overload. PMID- 11399555 TI - Erythropoiesis: Pyruvate Kinase Deficiency which Causes Nonspherocytic Hemolytic Anemia: The Gene and its Mutations. PMID- 11399556 TI - Malignancy: Current Clinical Practice: Current Therapeutic Options in Myelodysplastic Syndromes. AB - Myelodysplastic syndromes (MDS) are characterized initially by ineffective hematopoiesis and subsequently the frequent development of acute myelogenous leukemias (AML). During the last 15 years, important progress has been made in the understanding of the biology and prognosis of myelodysplastic syndromes. Risk adapted treatment strategies were established due to the high median age (60-75 years) of MDS-patients and the individual history of the disease (number of cytopenias, cytogenetical changes, transfusion requirements). The use of allogeneic bone marrow transplantation for MDS patients currently offers the only potentially curative treatment, but this treatment modality is not available for the most of the "typical" MDS-patients aged >60 years. Based on in-vitro findings analyzing the potential of several agents to differentiate or to stimulate hematopoietic progenitor cells a number of therapeutic options were evaluated in clinical trials: hematopoietic growth factors (e.g. erythropoietin, G-CSF), differentiation inducers (e.g. retinoids), or cytoprotective substances (amifostine). The role of immunsuppressive agents (antithymocyte globulin, cyclosporine A) either alone or in combination is being actively investigated. Using intensive cytotoxic treatment in patients with advanced MDS or AML after MDS complete remission rates comparable with those known from the treatment of de novo AML were reported. The therapy related toxicity (early death rate <10%) was reduced by using G-CSF given prior ("Priming") and/or after the cytotoxic treatment. PMID- 11399557 TI - Malignancy: A New Approach to the Analysis of Apoptosis in the Leukemic Subpopulation by Flow Cytometry Using a CD45 Gating Strategy. AB - Bone marrow and peripheral blood are heterogeneous tissues containing cells of different hematopoietic lineages. It is possible to detect leukemic cells by flow cytometry using a gating strategy, which combines CD45 expression on the cell surface with right angle light scatter (SS). This approach was applied to 15 cases of AML. Myeloblasts had the lowest CD45 fluorescence intensity of any of the cells in the myeloid series and also had the lowest SS, approximately equivalent to monocytes, but greater than lymphoblasts and lymphocytes. Using this gating strategy in each sample we could identify up to 5 separate cell compartments. Our results showed good correlation between the flow differential and the manual differential cell count. However in some cases, especially when a sample became hypocellular, the flow differential was more sensitive in identifying leukemic blasts. Total apoptosis (i.e. apoptosis in all cell populations combined) varied during the treatment between 0-34%. In the blood, the highest percentage of total apoptotic cells usually occurred between day 3-5 of treatment. The percentage of apoptotic cells varied depending on the cell type on a percentage basis. The leukemic population was lesslikely to undergo apoptosis compared to the lymphocytes, monocytes and more mature myeloid cells. In normal cells, apoptosis occurred mostly in G(1) and S phases of the cell cycle. Apoptosis among CD45-blasts usually varied between 0-5%. Myeloblasts also had a tendency to undergo apoptosis in G(1) and S phases of the cell cycle. The CD45-blast apoptotic peak in the blood occurred between day 5-7 of treatment. Analysis of drug-induced apoptosis in bone marrow seems to provide more information than such measurements in peripheral blood. PMID- 11399558 TI - Malignancy: Case Report: Muscle Involvement in Multiple Myeloma: Report of a Patient Presenting Clinically as Polymyositis. AB - Although bone pain is common in multiple myeloma (MM), muscular symptoms, especially myalgias, may be rare. We describe a patient who presented with generalised myopathy and elevated creatine kinase (CK) suggestive of polymyositis. Routine blood tests showed raised viscosity and marked rouleaux formation in the peripheral blood film. A serum protein electrophoresis showed IgG Lambda paraproteinemia with immunoparesis. A sternal bone marrow aspirate and a bone marrow biopsy concurrently obtained from the right posterior iliac crest showed considerable (15-20%) marrow infiltration with plasma cells confirming a diagnosis of multiple myeloma. A review of the literature suggests that generalised myopathy and elevated CK associated with MM have not been reported in the past. We believe this is the first reported case of MM presenting as polymyositis. PMID- 11399559 TI - Molecular Medicine and Molecular Pathology for Hematologists: II. Lets go Techno; Principles of the Molecular Technologies and their Applications in Haematology. AB - Molecular techniques have a key role to play in laboratory and clinical haematology. Restriction enzymes allow nucleic acids to be reduced in size for subsequent analysis. In addition they allow selection of specific DNA or RNA sequences for cloning into bacterial plasmids. These plasmids are naturally occuring DNA molecules which reside in bacterial cells. They can be manipulated to act as vehicles or carriers for biologically and medically important genes, allowing the production of large amounts of cloned material for research purposes or to aid in the production of medically important recombinant molecules such as insulin. As acquired or inherited genetic changes are implicated in a wide range of haematological diseases, it is necessary to have highly specific and sensitive assays to detect these mutations. Most of these techniques rely on nucleic acid hybridisation, benefitting from the ability of DNA or RNA to bind tighly to complimentary bases in the nucleic acid structure. Production of artificial DNA molecules called probes permits nucleic acid hybridiation assays to be performed, using the techniques of southern blotting or dot blot analysis. In addition the base composition of any gene or region of DNA can be determined using DNA sequencing technology. The advent of the polymerase chain reaction (PCR) has revolutionised all aspects of medicine, but has particular relevance in haematology where easy access to biopsy material provides a wealth of material for analysis. PCR permits quick and reliable manipulation of sample material and its ability to be automated makes it an ideal tool for use in the haematology laboratory. PMID- 11399560 TI - Erythropoiesis: Paroxysmal Cold Haemoglobinuria: A Clinico-Pathological Study of Patients with a Positive Donath-Landsteiner Test. AB - 52 patients (30 male, 22 female) with paroxysmal cold haemoglobinuria (PCH) were identified by critically examining the records of all cases with Donath Landsteiner antibodies seen over a 37 year period. Although ages ranged from 1-82 years, PCH was much commoner in young children; the median age at presentation was 5 and the peak incidence, 0.4 per year per 100,000 of the population at risk, was in the 4 years and under group. 44 patients had acute transient PCH, 3 chronic non-syphilitic PCH and 1 chronic syphilitic PCH; 4, in whom the positive Donath-Landsteiner tests were incidental findings, could not be classified. Acute PCH typically presented in young children as sudden onset of malaise, haemoglobinuria and pallor, often associated with mild jaundice - all 30 patients who were 13 or younger had this type. There was usually a history of a recent viral type infection, most commonly of the upper respiratory tract. The occurrence of acute PCH had no obvious relation to exposure to cold. Dramatic and rapid falls in haemoglobin level were common, often accompanied initially by relative or absolute reticulocytopenia. The illness was severe, but the prognosis was generally good and the majority of patients had completely recovered within one month, some requiring no treatment. In approximately 68% of patients, blood transfusion was needed; the P blood group was not taken into account, but the patients were kept warm throughout. Steroids (usually prednisolone) were given in many cases; but since there was no evidence to support their benefit, it was recommended that they were stopped as soon as the diagnosis was confirmed. Chronic non-syphilitic PCH was much rarer; the patients had a characteristic presentation of severe systemic symptoms (paroxysms) and haemoglobinuria brought on by exposure to cold. The clinical manifestations varied in intensity between individuals; at the extreme, severe debility was experienced over many years. Warmth and avoidance of cold were an effective treatment, though in a severely afflicted patient, an attack could be precipitated by relatively little exposure to cold. With chronic syphilitic PCH there was the added need to treat the specific infection. The direct antiglobulin test was almost always positive (50 out of 51 cases tested), with C3d coating the red cells. The Donath-Landsteiner antibodies were of IgG class, but this was rarely demonstrated unless direct antiglobulin tests were carried out at 4 degrees C. The antibodies showed the classical anti-P specificity in 27 of the 30 patients tested; other specificities were unusual. Although acting much better as haemolysins, Donath-Landsteiner antibodies could also cause weak agglutination at room temperature. This was paralleled in vivo by predominantly intravascular haemolysis with an extravascular component. Diagnosis was usually easy when PCH was suspected, though in some patients Donath-Landsteiner tests did not become positive until more sensitive techniques involving papainised red cells or two-stage procedures were employed. Of particular interest was the association in some cases with lymphoproliferative disorders, collagen disease, myelodysplastic syndrome, delayed haemolytic transfusion reaction and other types of autoimmune haemolytic anaemia. In one patient, an aetiological relationship was confirmed by a saline extract of lymphoma tissue behaving as a strong Donath-Landsteiner antibody with the same anti-P specificity as the serum. 4 patients had other types of autoimmune haemolysis concomitant with, but distinct from, the PCH; in 3 cases this was cold haemagglutinin disease and in one it was warm type autoimmune haemolytic anaemia. PMID- 11399561 TI - Erythropoiesis: Splenic Immunoglobulin Variable Region Genes Encoding Red Blood Cell Binding Fab Fragments in Autoimmune Hemolytic Anemia. AB - An absolute requirement for the V(H(4-34) ) immunoglobulin (Ig) variable (V) region heavy chain (V(H) ) gene has been demonstrated in pathogenic cold agglutinin autoantibodies. Investigation of IgG binding anti-Rhesus (Rh) alloantibodies provides further evidence of V gene restriction in red blood cell (RBC) binding antibodies and demonstrates that the V(H(4-34) ) gene used to form cold agglutinins may also encode RBC antibodies of varied specificities. We reasoned that a similar V gene restriction may be evident in the gene segments encoding IgG anti-RBC autoantibodies mediating autoimmune hemolytic anemia (AIHA). To further examine this question IgG Fab fragment phage display libraries were constructed from the spleen of a patient with AIHA. The index autoantibody appeared to have incomplete anti-C specificity and bound all panel RBCs except Rh null. The Fab fragment phage display libraries were therefore panned twice on CDE/CDe RBCs and binding phage were eluted. Binding of the phage displayed Fab fragments to RBCs was confirmed by immunoflourescence and flow cytometry. Specificity was confirmed by the absence of binding to Rh null cells, murine RBCs and to human peripheral blood lymphocytes. Molecular analysis of Ig V genes encoding the pan RBC binding Fab fragments revealed a relative V(H) gene restriction and evidence of somatic mutation. The V(H(3) ) family member V(H(26) ) was prominent in RBC binding Fabs. The V(H(3) ) family member hV3005 and the V(H(4) ) family DP-65 gene segments also encoded RBC binding Fabs. The J(H(4) ) gene segment was present in all binding clones. Varied kappa and lambda light chain (V(L) ) genes were identified by sequencing and no single light chain was prevalent. Three of the ten V(L) and two of the three V(H) identified by sequencing appeared to derive from germline genes previously noted to have RBC binding specificity. We conclude that splenic Ig V genes can encode pan RBC binding antibodies with specificities similar to autoantibodies found in AIHA and that V(H) gene segment utilization by these antibodies is derived from a limited pool of somatically mutated V(H) gene segments. PMID- 11399562 TI - Erythropoiesis: Short Report: Translation of Analysis Results between Serum Ferritin Assays, Ferritin RIA AmershamTM and Abbott AxSYMTM Ferritin. AB - The serum ferritin assays, Ferritin RIA Amersham(TM) and Abbott AxSYM(TM) Ferritin were compared in order to translate values from one assay to the other. Serum ferritin was analysed with both assays in 102 samples. Logarithmic transformation of the results was performed in order to stabilize the variance. The relationship between the untransformed values was most exactly expressed by a proportionality: AxSYM Ferritin = 0.873 * RIA Ferritin. Due to this proportionality, the numerical difference between the assays increases with the ferritin concentration, although the percentage difference between the assays remains constant. PMID- 11399563 TI - Current Clinical Practice: Hereditary Haemochromatosis: Recent Advances in Biology and Management. AB - Hereditary haemochromatosis (HH) is an autosomal recessive disorder leading to excessive absorption of dietary iron. The gene affected in the common variety has recently been identified near the HLA-A locus on chromosome 6 and designated HFE. The HFE mutation, C282Y, responsible for most cases of HH is found in as many as 8-18% of people of European descent; 6-32 per 1000 are therefore homozygous, but only a variable proportion accumulate enough iron to develop organ damage with the associated clinical manifestations. Clinical expression depends on the amount of absorbable iron in the diet, the amount of iron being lost (greater in women through menstruation and pregnancies) and the severity of the genetic defect. The excess iron can be removed by serial phlebotomies. In symptomatic individuals this reverses some of the manifestations and prolongs survival significantly; if treatment is instituted before symptoms appear all manifestations are prevented. The objective must therefore be to identify and treat affected individuals in the presymptomatic stage. In this context, the role of population screening is currently being debated. PMID- 11399564 TI - Bone Marrow Transplantation: Prognostic Factors of Peripheral Blood Stem Cell Mobilization with Cyclophosphamide and Filgrastim (r-metHuG-CSF): The CD34+ Cell Dose Positively Affects the Time to Hematopoietic Recovery and Supportive Requirements after High-Dose Chemotherapy. AB - To prospectively analyze factors that influence peripheral blood stem cell (PBSC) collection and hematopoietic recovery after high-dose chemotherapy (HDC), 39 patients received cyclophosphamide 4 g/m(2) and rHuG-CSF (Filgrastim) 5 &mgr;g/kg/day. Leukapheresis was started when CD34(+) cells/mL were > 5 x 10(3). A minimum of 2 x 10(6) CD34(+) cells/kg was collected. Median steady-state bone marrow CD34(+) cell percentage was 0.8% (range, 0.1 to 6). Thirty-two patients received HDC with autologous PBSC transplantation plus Filgrastim. A median of 2 (range, 0 to 6) leukapheresis per patient were performed and a median of 6.3 x 10(6) CD34(+) cells/kg (range, 0 to 44.4) collected; four patients failed to mobilize CD34(+) cells. The number of cycles of prior chemotherapy had an inverse correlation with the number CD34(+) cells/kg collected (r = -0.38; p < 0.005). Patients with <7 cycles had a higher predictability for onset of leukapheresis than patients with (3) 7 (93% versus 50%; p < 0.005). The four patients who failed to mobilize had received >/=7 cycles. The number of CD34(+) cells/kg infused after HDC had an inverse correlation with days to recovery to 0.5 x 10(9) neutrophils/L and 20 x 10(9) platelets/L (r = -0.68 and -0.56; p < 0.005). The effect of these factors on mobilization and hematopoietic recovery were confirmed by multivariate analysis. Requirements for supportive measures were significantly lower in patients given a higher dose of CD34(+) cells/kg. Therefore, PBSC collection should be planned early in the course of chemotherapy. Larger number of CD34(+) cells/kg determined a more rapid hematopoietic recovery and a decrease of required supportive measures. PMID- 11399565 TI - Malignancy: Molecular Demonstration of BCR/ABL Fusion in a Patient with Chronic Myelogenous Leukemia with Basophilia Carrying a Variant t(16;22) (q24;q11) Philadelphia Chromosome. AB - We report a patient with chronic myelogenous leukemia in chronic phase and basophilia which was found to carry a simple variant t(16;22) (q24;q11) Philadelphia (Ph) chromosome in unstimulated bone marrow mononuclear cells. Molecular analysis of peripheral blood and bone marrow mononuclear cells demonstrated the presence of a bcr-abl chimeric mRNA transcript of the b(3) -a(2) type. These findings confirm that band 9q34 participates in the formation of all Ph chromosomes, either standard or variant, even when this is not detectable by conventional cytogenetics. The available literature concerning variant Philadelphia translocations is also reviewed. PMID- 11399566 TI - Malignancy: Insulin-like Growth Factor-1 (IGF-1) is a Costimulator of the Expansion of Lineage Committed Cells Derived from Peripheral Blood Mobilized CD34+ Cells in Multiple Myeloma Patients. AB - The effect of insulin-like growth factor-1 (IGF-1) on highly enriched human apheresis CD34(+) progenitor cells was investigated in vitro. The progenitor cells were mobilized by treatment with cyclophosphamide + granulocyte - colony stimulating factor (G-CSF) in patients with multiple myeloma. CD34(+) cells were cultured for 7 days in serumfree medium containing stem cell factor (SCF), granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-3 (IL 3), and this is referred to as cytokine-dependent proliferation. After 7 days of cytokine-dependent proliferation the total number of viable cells increased 1.6 8.2 times, and subsets of cells expressing the granulocyte marker CD15, the myelomonocytic marker CD64 and the erythrocyte phenotype CD71(high) /CD64(-) were detected among the in vitro cultured cells. Addition of G-CSF together with SCF + IL-3 + GM-CSF increased the number of CD15(+) and CD64(+) cells, but without altering the number of erythroid cells. IGF-1 caused a dose-dependent increase in the number of CD15(+), CD64(+) and CD71(high) /CD64(-) cells, and this increase was detected when cells were cultured in both SCF + IL-3 + GM-CSF alone and G-CSF + SCF + IL-3 + GM-CSF. A minor subset of CD34(+) cells could still be detected among in vitro cultured cells and the number of CD34(+) cells was not altered by adding G-CSF and/or IGF-1. Morphologically recognizable mature granulocytes or erythroid cells could not be detected for any of the combinations investigated. We conclude that IGF-1 can enhance the in vitro proliferation of committed progenitor cells derived from apheresis CD34(+) cells. PMID- 11399567 TI - Malignancy: The Number of Nucleoli and Main Nucleolar Types in Lymphoblasts of Children Suffering from Acute Lymphoid Leukemia. AB - Nucleoli were studied in lymphoblasts of children (untreated with cytostatic therapy) suffering from acute lymphoblastic leukemias (ALL) by means of a simple cytochemical procedure for the demonstration of RNA to provide information on the incidence of the main nucleolar types and the number of nucleoli in these cells. The values of the nucleolar coefficient reflecting the number of nucleoli per lymphoblast ranged between 1.66 and 2.03. The slightly larger values of the nucleolar coefficient in T-ALL were not statistically significant in comparison with those in nonT-ALL. Lymphoblasts in the bone marrow as well as the peripheral blood of both nonT and T-ALL patients mostly contained, "active" (RNA transcribing) large nucleoli with a relatively uniform distribution of RNA. "Inactive" micronucleoli or particularly "resting" ring shaped nucleoli were noted less frequently in these cells. On the other hand, the larger percentage of lymphoblasts determined in specimens stained with the panoptic staining (May Grunwald-Giemsa) in comparison with that of lymphoblasts with "active nucleoli" in specimens stained for RNA apparently indicates the absence of such nucleoli in some of these cells. These cells might represent ageing, not proliferating, cells the existence of which has been already suggested by previous studies based on a different methodical approach. PMID- 11399568 TI - The Hematopoietic Microenvironment: Stromal Cell Types: Characterization and Function In Situ and In Vitro. AB - The murine hematopoietic stroma is essential for the homing and the continued replication of hematopoietic stem cells following transplantation and appears to control their lineage proliferation. In our studies it has been found to consist of 3 cell types with differing antigenic displays, growth factor responses and interactions when placed in culture. These characteristics are in keeping with their classifications as myofibroblasts, endothelial like cells and macrophages, respectively. Cell adhesion molecules (CAMs) and early acting cytokines have been found to be associated with these cell types. Only the myofibroblast type has been found to support proliferation of the descendants of long term repopulating stem cells (LTRC) plated over their confluent layer. The endothelial like cell type, and possibly the macrophage, were found to suppress not only the growth of LTRC but, also, that of the myofibroblasts obtained from the same marrow samples. Culture medium obtained from near-confluent growth of the endothelial like cells contained a growth inhibiting molecule of less than 3000 da. capable of inhibiting cells of several types, including the stromal myofibroblasts. The endothelial like cells were shown to unilaterally express VCAM-1 in culture, as well as vWf and endothelial cell specific antigens. Tissue sections of femurs from normal irradiated recipients taken 1 hour post-transplantation revealed Lac Z marked donor stem cells lodged just external to the endothelial lining of thin walled vessels and in contact with myofibroblasts. Previous studies had shown that some stem cells enter replication at these sites within 48 hours of lodgment. It is suggested that a balance of growth stimulation versus suppression may be maintained between the myofibroblasts and the endothelial like cell types, respectively, in the functioning marrow stroma. Both the myofibroblastoid and the endothelial like stromal cells of old mice displayed a reduced replicative rate in vivo and a reduced replicative capacity in vitro. This suggests that treatments that induce damage to the marrow microenvironment may be especially destructive in aged individuals. In recent experiments it has become clear that the stromal cell population that includes cells identifiable as alkaline phosphatase positive myofibroblasts also contains cells that produce osteocalcin, osteonectin and bone sialoprotein, and deposit von Kossa positive calcium. Whether this results from a single cell lineage or two lineages of similar morphology and alkaline phosphatase expression is under investigation. PMID- 11399569 TI - The Hematopoietic Microenvironment: Phenotypic and Functional Characterization of Human Marrow Vascular Stromal Cells. PMID- 11399570 TI - Malignancy: Current Clinical Practice: Treatment of Hairy Cell Leukemia at the Close of the 20th Century. AB - In the last half of this century, hairy cell leukemia was recognized as a distinct B-cell malignancy, accounting for 2% of all leukemias. Characteristics include splenomegaly, pancytopenia, a usually indolent course, and responsiveness to both interferon and purine analog therapy. Accurate diagnosis requires the demonstration of malignant cells in the bone marrow and peripheral blood which contain cytoplasmic projections and characteristic surface antigens. Splenectomy was identified early as a palliative therapy, and in 1984 systemic treatment with interferon alpha was first reported to induce complete remissions. Soon thereafter, the purine analog deoxycoformycin was found to induce more durable complete remissions in a higher percentage of patients. In 1990, 2 Chlorodeoxyadenosine, a new purine analog therapy, was reported to be capable of inducing long-term durable responses in most patients after a single cycle. Current challenges include identifying which purine analog is the least toxic since both appear similarly effective, and neither appear to add to the already increased rate of second malignancies occurring in these patients. Moreover, up to 25% of patients with hairy cell leukemia fail initially or eventually to respond to standard therapy, making the development of new approaches necessary. The characteristic bright expression of several B-cell antigens on the malignant cells, including CD20, CD22 and CD25, has led to the development of targeted biotherapeutic approaches. A recombinant immunotoxin targeting CD25 has recently been reported to induce major responses and it is likely that other successful targeted approaches will be reported early in the new century. PMID- 11399571 TI - Malignancy: Granulocyte Colony Stimulating Factor Increases the Efficacy of Conventional Amphotericin in the Treatment of Presumed Deep-Seated Fungal Infection in Neutropenic Patients following Intensive Chemotherapy or Bone Marrow Transplantation for Haematological Malignancies. AB - A prospective, comparative study of empiric amphotericin B with, or without, granulocyte colony stimulating factor was carried out to assess whether the addition of granulocyte colony stimulating factor to empiric amphotericin B improves the clinical response in neutropenic patients with suspected or proven fungal infection. Fifty nine neutropenic adults with haematological malignancy and antibiotic-refractory fever or clinical evidence of deep-seated fungal infection were studied. Patients received intravenous colloidal amphotericin B (1 milligram per kilogram body weight) with or without subcutaneous granulocyte colony stimulating factor (three to five micrograms per kilogram body weight). Thirty patients received amphotericin alone and 29 amphotericin plus granulocyte colony stimulating factor. Nearly twice as many patients responded to amphotericin B with concomitant administration of granulocyte colony stimulating factor (62%) as responded to amphotericin alone (33%; difference in proportions 0.29, 95%CI 0.03-0.54). Clinical response in patients receiving granulocyte colony stimulating factor coincided with neutrophil recovery in most cases. Addition of granulocyte colony stimulating factor to empiric amphotericin B significantly reduced the number of patients requiring salvage therapy with lipid associated or liposomal formulations of amphotericin B addition of granulocyte colony stimulating factor to empiric intravenous amphotericin B improves the response rate and thereby reduces the number of patients requiring salvage therapy with liposomal or lipid-associated preparations of amphotericin B. PMID- 11399572 TI - Malignancy: X-Linked Cytochrome B Positive Chronic Granulomatous Disease treated by Bone Marrow Transplantation. AB - Chronic granulomatous disease is caused by a genetic defect in the oxidase of phagocytic cells which results in increased susceptibility to recurrent infections. Conventional treatment includes the use of antimicrobials and interpheron-gamma. This study was performed to assess the clinical efficacy of allogeneic bone marrow transplantation in definitively correcting the functional underlying defect of chronic granulomatous disease. An 8-year-old boy with a rare type X-linked cytochrome b positive chronic granulomatous disease underwent allogeneic bone marrow transplantation after conditioning with busulfan and cyclophosphamide. The patient's HLA identical sister was marrow donor. The post transplant outcome was uneventful. During the 9 year follow-up period the patient has been constantly free of infections, maintained an excellent clinical performance with full correction of the granulocyte functional defect. This case confirms that allogeneic bone marrow transplantation is the only treatment capable to cure chronic granulomatous disease to those patients who cannot be optimally treated with conventional therapy. PMID- 11399573 TI - The Hematopoietic Microenvironment: Stromal Extracellular Matrix Components As Growth Regulators For Human Hematopoietic Progenitors. PMID- 11399574 TI - Growth Factors: Thrombopoietic Property of the Pineal Hormone Melatonin. AB - The recent advances in the psychoneuroimmunology have suggested that the hematopoiesis may be under a neuroendocrine regulation, mainly exerted by the pineal gland. In particular, melatonin (MLT), which is the main pineal hormone, has appeared to stimulate platelet generation, probably by promoting the megakaryocyte fragmentation and modulating the cytokine network involved in platelet production. On this basis, we have evaluated the effect of pharmacological doses of MLT on platelet number in patients with persistent thrombocytopenia due to different causes. The study included 200 patients, who were randomized to receive supportive care alone or MLT at 20 mg/day orally in the evening, for al least 1 month. No MLT-related toxicity occured. Platelet mean number rapidly and significantly increased in response to MLT, and a normalization of platelet number was achieved on MLT therapy in 71/98 (72%) patients. The least responsive form of thrombocytopenia was DIC. This study shows that the pineal hormone MLT may represent a well tolerable and effective therapy of thrombocytopenia due to different pathogenetic mechanisms. PMID- 11399575 TI - Growth Factors: Production of Monocyte Chemotactic Protein-1 (MCP-1/JE) by Bone Marrow Stromal Cells: Effect on the Migration and Proliferation of Hematopoietic Progenitor Cells. AB - Recombinant chemotactic cytokines (chemokines) have been shown to modulate in vitro proliferation of hematopoietic progenitor cells. Whether bone marrow stromal cells produce chemokines and the physiological role they may have in the regulation of hematopoiesis has largely remained unexamined. We have examined the expression of monocyte chemoattractant protein-1 (MCP-1/JE) in bone marrow stromal cells and its effect on the migration and proliferation of murine hematopoietic progenitor cells. Freshly derived murine bone marrow stromal cells were found to secrete abundant amounts of MCP-1/JE, which was further increased upon stimulation of stromal cells with pro-inflammatory agents LPS, IL1-alpha, IFN-gamma, or TNF-alpha. Although culture supernatant conditioned by stromal cells exhibited chemotactic activity toward hematopoietic progenitor cells, the chemotactic activity was not due to MCP-1/JE. Furthermore, rMCP-1/JE also failed to induce migration of progenitor cells. MCP-1/JE, however, caused 20 to 30% increase in the clonal expansion of progenitor cells. Thus, although MCP-1/JE does not chemoattract hematopoietic progenitor cells it may have a role in their proliferation and clonal expansion. PMID- 11399576 TI - Erythropoiesis: Case Report: Congenital Dyserythropoietic Anemia Type II in a Woman Presenting with Jaundice, Anemia, and Splenomegaly. AB - Congenital dyserythropoietic anemias (CDAs) are extremely rare types of hemolytic anemias that share similar morphological findings and are characterized by ineffective erythropoiesis. CDAs are divided into three major groups and few variants. The most frequently encountered type is CDA type II (HEMPAS: Hereditary erythroblastic multinuclearity associated with a positive acidified serum test). We herein report a case of CDA type II, who presents with a mild anemia, jaundice, splenomegaly, cholelithiasis and hemolysis. CDA type II, about 120 cases have been reported so far, has recently been discovered to be due to the defective glycolization of membrane proteins on the erythrocyte progenitors. The responsible gene has been found to be located on the Chromosome 20q only a few years ago. PMID- 11399577 TI - Erythropoiesis: Hereditary Spherocytosis in Greece: Collective Data on a Large Number of Patients. AB - This paper summarizes data from a haematological, biochemical, and clinical study carried out in 73 patients of Greek origin (38 non-splenectomized children and 35 adults; 17 splenectomized) with Hereditary Spherocytosis (HS). Mean haemoglobin levels in the non-splenectomized patients were significantly lower (122 +/- 15 g/L) than those of the splenectomized group (144 +/- 15 g/L). In all patients with HS (non-splenectomized and splenectomized adults, and children) the MCHC values (355 +/- 22, 358 +/- 16 and 356 +/- 16 respectively) were significantly increased compared to a control group, while the percentage of microcytic and hyperchromic red cell subpopulations was significantly increased in the former group of adults. SDS-PAGE demonstrated that 29 patients (39.7%) had isolated spectrin deficiency, 22 patients (30.1%) had combined spectrin and ankyrin deficiency, 17 patients (23.3%) had band 3 deficiency and 1 patient had protein 4.2 deficiency. No quantitative biochemical defects were detected in 4 patients (5.5%). The biochemical findings did not correlate with the haematological and clinical phenotype of the disease. PMID- 11399578 TI - Hemostasis and Thrombosis: Clinical Usefulness of Combined use of Platelet Aggregation Test and Anti PF4-H. Antibodies Elisa test for the Diagnosis of Heparin Induced Thrombocytopenia. AB - Background. Heparin is the most commonly used drug in patients requiring therapeutic anticoagulation. But the use of heparin has serious side effects such as heparin-induced thrombocytopenia (HIT). The diagnosis of HIT is often difficult. This study was designed to test the diagnosis value of 2 laboratory tests: the platelet aggregation test and the ELISA test (Enzym Linked Immuno Sorbent Assay). Methods and Results. In order to evaluate the diagnostic value of these 2 tests, we prospectively performed both tests in all patients referred to our laboratory with suspected type II HIT, and compared their results with clinical outcome. Plasmas from 60 patients suspected of HIT, were tested for heparin-dependent platelet-reactive antibodies with a platelet agregation test (PAT) and with an ELISA which detects anti heparin platelet factor 4 antibodies (Anti PF4-H Ab). Among the 60 explored patients, the clinical diagnosis of HIT was confirmed in 27 patients by clinical criteria. In 16 of these 27 patients, PAT and anti PF4-H. Ab were both positive, and in the 11 other patients, one of the 2 tests was negative. In 29 of the 33 patients with no clinical HIT, PAT and anti PF4-H. Ab were both negative, in the 4 remaining patients, one of the 2 tests was positive. Conclusion. For 75% of patients, biological results were concordant with the final clinical diagnosis. The combination of both tests is more reliable than the use of a single test; in the present series, all patients with positive results on both tests had clinically confirmed HIT, and all patients with negative results on both tests had not clinically confirmed HIT. PMID- 11399579 TI - Haemostasis and Thrombosis: Current Clinical Practice: Low-Molecular-Weight Heparins in The Prophylaxis and Treatment of Thrombo-Embolic Disease. AB - Low-Molecular-Weight Heparin (LMWH) fractions are prepared from standard unfractionated heparin (UFH) and are thus similar to it in many aspects. The major advantages of LMWH are improved efficacy and safety, longer half-life and reduced need for laboratory monitoring. In addition, the dangers of UFH administered by continuous infusion in the hospital setting are often not fully appreciated and the necessary monitoring and dosage adjustment poorly carried out resulting in inadequate doses being given. LMWHs are the drug of choice in many clinical situations. Four LMWHs are now licensed in the UK for prophylaxis of venous thrombo-embolism during or after surgery (Certoparin, Dalteparin [Fragmin], Enoxaparin [Lovenox/Clexane] and Tinzaparin [Innohep]; a fifth is licensed but not currently available in the UK. Dalteparin, Enoxaparin and Tinzaparin are licensed for the treatment of Deep Vein Thrombosis (DVT), and Tinzaparin additionally for the treatment of Pulmonary Embolism (PE), but so far none is licensed for use in pregnancy or paediatrics. PMID- 11399580 TI - Myeloproliferative Disease: Polycythemia Vera: The Packed Cell Volume and The Curious Logic of The Red Cell Mass. AB - Since the first systematic blood volume studies of polycythemia in the 1920s, measurement of blood volume and red cell mass (RCM) has become routine. However, the radionuclide-labeling methods promulgated by the International Committee for Standardization in Haematology (ICSH) remain complex and poorly understood. Many hematologists and other clinicians err in the belief that these methods permit "direct measurement" of RCM, whereas the ICSH method is indirect: it requires calculation of RCM from (PCV) x (whole blood volume). The use of an elevated value of PCV to calculate RCM in order to evaluate the same elevated value of PCV is a curiously circular logic that is embraced by most clinicians and most hematologists. Analysis of published data in 186 cases of polycythemia vera indicates that RCM is an exponential function of PCV. In most cases, PCV alone suffices to document normal or increased RCM. Relative polycythemia results from dehydration, not from stress. Clinicians need to be aware of the range of physiologic fluctuations that normally occur in plasma volume. Realistic criteria for normal ranges of PCV, Hb concentration and RCM should be adopted in clinical laboratories so that clinicians will not be misled to undertake futile and costly investigations of results that are in the upper percentiles of the normal distribution, as exemplified by the Ulysses Syndrome. PMID- 11399581 TI - Myeloproliferative Disease: Markers of Endothelial and Platelet Status in Patients with Essential Thrombocythemia and Polycythemia Vera. AB - Vascular complications are the main cause of morbidity in polycythemia vera (PV) and essential thrombocythemia (ET). To investigate plasma concentrations of soluble P-selectin (sP-Sel.), soluble E-selectin (sE-Sel.) and soluble thrombomodulin (sTM) in relation to the presence of thromboembolic events 38 patients with Chronic Myeloproliferative Disorders (CMD) (14 PV pts and 24 ET pts), 15 age - matched controls and 15 patients with secondary thrombocytosis were studied. Plasma levels of P-Sel., E-Sel. and TM were significantly increased in the group of patients as compared with control subjects (respectively p < 0.001, p < 0.04 and p < 0.01). sP-Sel. levels showed no significant difference between the patients and those with secondary thrombocytosis. No difference in sP sel levels were also observed between subgroups of CMD patients with and without vascular complications. However, among patients with ET, those with thrombosis had higher sP-Sel levels than those without thrombosis (1.177 +/- 110.48 ng/ml vs 816.25 +/- 99.27 ng/ml). High levels of sE-Sel and sTM were found in CMD patients (71.93 +/- 39.08 ng/ml and 35.81 +/- 20.79 ng/ml, respectively). Plasma sE-Sel. concentration was significantly higher in CMD patients with thrombosis than that in CMD patients without thrombosis (p < 0.001). There was no difference in sTM concentration between two groups. These findings indicate that sustained endothelium and platelet activation is present in patients with ET and PV and it might contribute to the pathogenesis of thromboembolic events in these patients. PMID- 11399582 TI - Malignancy: 2'-Chlorodeoxyadenosine Effectively Induces Complete Remission in Hairy Cell Leukaemia. AB - Hairy cell leukaemia, previously known as leukaemic reticuloendotheliosis, is an indolent lymphoproliferative disorder of unknown etiology. It typically affects males, causes marked splenomegaly and moderate enlargement of the liver, whilst lymphadenopathy is inconspicuous. Pancytopenia is characteristic with unusually profound monocytopenia, variable reduction in platelets, and the presence in the peripheral blood and marrow of abnormal small lymphocytes having irregular cytoplasmic margins. Ultrastructure, combined with cytochemistry and flow cytometry, have refined diagnosis. A variant exists between this classical entity and B prolymphocytic leukaemia, where blastic transformation or massive lymph node enlargement are found, and this is of ominous significance. In all these patients with this entity conventional chemotherapy is ineffective and shortens survival. Our previous experience with splenectomy results in excellent clinical control for long periods of time, but without disease eradication. There followed a vogue for the use of interferon but this is limited by high cost and dose dependent side-effects. Contemporary management centres on the purine analogues, where durable responses are possible with fludarabine and deoxycoformycin, but best with 2'chlorodeoxyadenosine (2-CDA). To document the efficacy of the latter agent, we analysed the outcome in seventeen consecutive patients treated over the last five years. Four were ineligible for analysis, although two had 2-CDA. The other thirteen, managed on a standard seven-day course of 0.1 mg/kg 2-CDA given as a continuous intravenous infusion, all responded promptly. Apart from transient leucopenia complications have been minimal, and oral co-trimoxazole prophylaxis for pneumocystis carinii was maintained during the first one year. In all thirteen there was a rapid return to normal of peripheral blood count and marrow on aspiration and trephine biopsy. Even in the longest follow-up clinical and haematologic remission has been maintained and no patients have required retreatment. One individual has relapsed in the marrow at two years. Despite the relative expense of the agent the excellent treatment outcome and patient acceptability, coupled with its safety, leads to the recommendation that in South Africa - as elsewhere in the world - this be regarded as the first line of treatment. PMID- 11399583 TI - Malignancy: Case Report: Acute Promyelocytic Leukemia in Late Pregnancy. Successful Treatment with All-Trans-Retinoic Acid (ATRA) and Chemotherapy. AB - The use of all-transretinoic acid (ATRA) in APL is a great advance in the treatment of acute leukemia, driving the maturation steps until adult form. The effect of this medication in pregnant women with APL is being a safe and effective treatment only after the first trimester of pregnancy. If used in the first three months, it can cause fetus malformations due to its potent teratogenicity. The two patients we reported here, gave birth to normal children, yet they received ATRA. After a week with ATRA treatment, fibrinogen level improved and "D" dimers decreased, so as observed slowly maturation of leukemic leucocytes. ATRA used at the same time with chemotherapy; such as Cytarabine and Idarrubicine seems highly useful for induction and long term control of disease. One short discussion about the findings and compare with those found in the literature are presented. PMID- 11399584 TI - The Hematopoietic Microenvironment: Osteoblasts and The Hematopoietic Microenvironment. AB - Cells of the bone marrow are chiefly dedicated to two processes; the production of blood, and the production of bone that houses the hematopoietic organ. The majority of our understanding of these processes comes from data focused on one of these functions. Yet, in vivo the processes are intermixed. Our recent data demonstrate that human osteoblast-like cells have important accessory roles in hematopoiesis. These data include the demonstration that human osteoblast-like cells; support the growth of primitive human hematopoietic progenitors (CD34(+) cells) in short and long term cultures and, synthesize multiple cytokines believed to regulate hematopoiesis. Based upon anatomic and developmental findings characterizing hematopoietic cells in close approximation with endosteal cells, and these findings, we hypothesize that osteoblasts play a critical role in hematopoietic cells development in vivo. PMID- 11399585 TI - The Hematopoietic Microenvironment: The Osteogenic Compartment of Bone Marrow: Cell Biology and Clinical Application. AB - The marrow stromal cells of the bone cavities are of mesenchymal origin. These cells are maintaining bone remodeling and control the differentiation of hemopoietic cells during the life span. The function of the marrow stroma is based on cells that differentiate into various subtypes that are derived from a common stem cell. Plating the bone marrow at low density, they form colonies with a fibroblast-like appearance (CFU-F). The fibroblastic colonies may differentiate to distinct cell types as fibroblasts, endothelial, osteogenic and adipocytes. Understanding of the control mechanism of these cells' differentiation is very important. We used an in vitro system from established primary cultures and continuos cell lines, MBA cells from mouse bone marrow. The MBA cells were used to study the stromal cells subtypes. Herein, we summarized the knowledge on the osteogenic cells' differentiation using the marrow-derived stromal osteogenic cells, MBA-15 and clonal lines. The characterization of cells was based on morphology, rate of cell growth, gene expression and biochemical profile. A deeper understanding of the osteogenic cells differentiation will lead to better use of the marrow stroma cells in clinical application and tissue engineering to ensure healing of bone defects or skeletal genetic diseases. PMID- 11399586 TI - Erythropoiesis: Review Article: Slow and Steady Wins The Race? Progress in the Development of Vectors for Gene Therapy of beta-Thalassemia and Sickle Cell Disease. AB - The cloning of the human beta-globin genes more than 20 years ago led to predictions that beta-thalassemia and sickle cell disease would be among the first monogenic diseases to be successfully treated by gene replacement therapy. However, despite the world-wide enrollment of more than 3,000 patients in approved gene transfer protocols, none have involved therapy for these diseases. This has been due to several technical hurdles that need to be overcome before gene replacement therapy for beta-thalassemia and sickle cell disease can become practical. These problems include inefficient transduction of hematopoietic stem cells and an inability to achieve consistent, long-term, high-level expression of transferred beta-like globin genes with current gene transfer vectors. In this review we highlight the relationships between understanding the fundamental mechanisms of beta-globin gene locus function and basic vector biology and the development of strategies for beta-globin gene replacement therapy. Despite slow initial progress in this field, recent advances in a variety of critical areas provide hope that clinical trials may not be far away. PMID- 11399587 TI - Erythropoiesis: Erythrocyte Deformability is Reduced and Fragility increased by Iron Deficiency. AB - Hypochromic and microcytic erythrocytes, resulting from absolute iron deficiency, have a shortened survival due to ineffective production coupled with accelerated reticuloendothelial cell sequestration once released into the circulation. To explore the contribution of diminished deformability and increased fragility to these in vivo phenomena, iron status was correlated with the corresponding in vitro measurements using ektacytometry on whole erythrocytes or resealed ghost membranes. Compared to normal controls (Group 1: n = 15), those with iron deficiency of unknown origin (Group 2: n = 15), or when associated with primary proliferative polycythemia (Group 3: n = 20), were significantly less deformable and more fragile, whereas the latter category of patients, who were not anaemic (Group 4: n = 33), occupied an intermediate position. These observations support the contention that shortened intramedullary and extramedullary lifespan is contributed to by the altered physical characteristics of the red cell membrane. This observation is therapeutically important since the lesion can be readily corrected by oral iron supplementation which, at the same time, reverses the symptoms of the anaemia and prevents the development of paradoxical hyperviscosity. Additionally, it is speculated that the risk of thrombotic events in the polycythemic group may be reduced, since these appear to occur more frequently in the face of depleted body iron stores. PMID- 11399588 TI - Haemostasis and Thrombosis: Current Clinical Practice: Current Management of Thrombotic Thrombocytopenic Purpura. AB - Historically, the mortality rate of thrombotic thrombocytopenic purpura (TTP) approached 100%. However, by the 1980's, new therapy was instituted with a vast improvement in survival to 90%. The exact pathogenesis of TTP remains elusive. Yet, despite incomplete understanding of the pathophysiology, outcome has improved due to increased awareness of the symptomatology leading to earlier diagnosis and better supportive care, in addition to effective therapy with plasma exchange. TTP represents a disease in which prompt diagnosis and treatment can lead to a critical difference in clinical outcome. PMID- 11399589 TI - Malignancy: Idiotypic Immune Targeting of Multiple Myeloma. AB - Immunotherapy of malignant diseases remains a major therapeutic challenge. In multiple myeloma, interests have been focussed primarily on the targeting of the idiotypic protein since it represents one of the few tumor specific antigens. Various vaccination strategies have been employed, including the use of naked protein, protein conjugated to Keyhole Limpet Hemocyanin and, most recently, vaccine delivered via dendritic cells. Studies so far indicate the feasibility of inducing potentially beneficial idiotype-specific immune responses in the autologous hosts. However, the antitumor responses produced by the vaccines have been generally modest and sup-optimal. Further work is therefore ongoing to optimize the antitumor responses by including cytokines in the vaccination schedule. In addition, the development of a polyvalent vaccine is also being explored. The next few years should prove interesting and crucial in bringing effective and reliable immunotherapeutic strategies to the clinics. PMID- 11399590 TI - Malignancy: Adjuvant Radiotherapy to Initial Bulky Disease in Patients with Advanced Stage Hodgkin's Disease. AB - To determine if the use of adjuvant radiotherapy to sites of initial bulky disease and adequate modern chemotherapy in patients with advanced stages (IIIB and IV) Hodgkin's disease could improve duration of remission and overall survival. Patients previously untreated with pathologically documented advanced stages Hodgkin's disease were randomly assigned to received chemotherapy alone with EBVD regimen (epirubicin, bleomycin, vinblastine and dacarbazine): 56 patients or combined therapy: The same chemotherapy regimen following by adjuvant radiotherapy (35 Gy) to sites of initial bulky disease (tumor mass >7 cm diamenter): 54 patients. Five year overall survival rates were 88% (48 patients) and 60% (34 patients) from combined therapy compared to chemotherapy alone respectively (p < 01) (95% confidence interval (CI): for the difference 18% to 39%). Five-year failure free survival were 83% and 50% respectively (p < 01) (95% CI for difference: 22% to 35%). Toxicity was moderate and well tolerate. No death related treatment were observed. After a median follow-up of 66 months, no second solid neoplasmas or acute leukemia has been observed. The use of adjuvant radiotherapy to sites of initial bulky disease following the use of modern chemotherapy in patients with advanced stages Hodgkin's disease improve outcome with increase in failure free survival and overall survival, with moderate toxicity. More randomized clinical trials are warranted to define this therapeutic approach. PMID- 11399591 TI - Malignancy: Case Report: Hypereosinophilia Progressing to Granulocytic Sarcoma and Acute Myelocytic Leukemia with Trisomy 8: A Case Report and Review of the Literature. AB - Conditions associated with increased peripheral blood and bone marrow eosinophil count may be reactive, clonal or idiopathic. Clonal eosinophilic disorders are characterized by increased production of eosinophils alongside a clone of malignant cells. In these patients, the eosinophils can either be demonstrated as being part of the malignant clone or produced as a result of cytokine production by the malignant clone. Criteria for the diagnosis of idiopathic hypereosinophilic syndrome (HES) include the exclusion of other known causes of hypereosinophilia. A few patients with the initial diagnosis of HES develop clonal disorders manifested by granulocytic sarcoma or acute leukemia. We report a patient with a nine year history of HES before progressing to chloroma and acute leukemia. Cytogenetic studies on the bone marrow specimen revealed trisomy 8. This report and others in the literature support the concept that at least some cases of HES are as yet unidentified clonal diseases. Cytogenetic studies are therefore recommended at diagnosis and during the follow up of patients with HES. PMID- 11399592 TI - Malignancy: Case report: Myelodysplasia Following Treatment of Chronic Lymphocytic Leukaemia with Fludarabine. AB - Recently there has been an increased awareness of a possible link between the use of purine nucleoside analogues and myelodysplasia. We report the case of a patient who developed myelodysplasia with complex cytogenetic changes after receiving fludarabine. We review the literature, discuss the possible links between myelodysplasia and nucleoside analogues and putative mechanisms for secondary neoplasia. PMID- 11399593 TI - BMT: Fractionation of Gene Modified Hematopoietic Autografts into Multiple Weekly Infusions Does Not Improve Engraftment in Unconditioned Canine Recipients. AB - Hematopoietic stem cell (HSC) gene therapy will require efficient transfer of genes to HSCs and long term engraftment and proliferation of genetically modified HSCs following adoptive transfer. We evaluated whether fractionation of grafts into 4-5 weekly infusions to non-myeloablated, autologous canine recipients would improve engraftment of genetically modified HSCs. Experimental animals and controls receiving a single infusion had similar levels of engraftment with approximately 3-10% of marrow derived progenitors carrying transgene sequences for up to 29 months. There appears to be no improvement of engraftment of genetically modified HSCs in non-myeloablated large animal recipients by dose fractionation. PMID- 11399594 TI - BMT: Serum Ferritin as Risk Factor for Veno-occlusive Disease of the Liver. Prospective Cohort Study. AB - Veno-occlusive disease of the liver (VOD) is an important complication in hematological transplantation. The aim of this study is to analyze the risk factors for VOD and other forms of liver toxicity in a cohort of 180 peripheral stem cell transplants performed in our Center. We find that elevated pretransplant levels of serum ferritin are the most important risk marker for VOD. We believe that ferritin reflects damage induced by oxygen radicals resulting from iron-mediated catalysis. We also discuss different risk factors for VOD and other forms of liver toxicity, suggesting diferent pathogenic mechanisms. PMID- 11399595 TI - The Hematopoietic Microenvironment: Matrix Metalloproteinases in the Hematopoietic Microenvironment. AB - Matrix metalloproteinases (MMPs) are structurally and functionally related zinc dependent endopeptidases capable of degrading the components of extracellular matrix (ECM) and basement membranes. MMPs participate in many physiological processes and have also been implicated in various pathological conditions including tumor invasion and metastasis. The functions of MMPs are known to be controlled by mechanisms leading to activation of their latent forms and through inhibition of both active and latent forms by natural tissue inhibitors of metalloproteinases (TIMPs). The complex relationships between MMPs and TIMPs within the bone marrow microenvironment during normal hematopoiesis as well as during leukemic growth and dissemination have not been extensively investigated. We reported that primary acute myelogenous leukemia (AML) blasts and leukemic KG 1 cells penetrate reconstituted basement membrane (Matrigel) in an in vitro invasion assay, secrete the gelatinases (MMP-2 and MMP-9) and express active MMP 2 on the cell surface. We also analyzed MMP/TIMP expression in normal bone marrow cells of the myeloid and stromal lineages and showed that MMP-2, MMP-9, TIMP-1 and TIMP-2 are produced in the bone marrow microenvironment. Furthermore, we examined the role of gelatinases in the transmigration of stem/progenitor cells from the bone marrow into peripheral blood. We found that steady-state bone marrow CD34(+) cells, unlike circulating peripheral blood CD34(+) cells, did not express MMP-2 and MMP-9 mRNA transcripts and proteins, and that various cytokines were able to upregulate expression of these MMPs in bone marrow CD34(+) cells and trans-Matrigel migration of these cells. Thus, we now have evidence that MMPs and TIMPs are constituents of the hematopoietic microenvironment although their roles in hematopoiesis have yet to be determined. PMID- 11399596 TI - Erythropoiesis: Correlations Between Iron Status Markers During Normal Pregnancy in Women with and without Iron Supplementation. AB - The aim was to evaluate relationships between iron status markers (haemoglobin, erythrocyte indices, serum iron, serum transferrin, serum transferrin saturation, serum ferritin) in normal pregnancy. Iron status markers were measured at 4-week intervals during pregnancy and postpartum in 120 healthy women; 62 had daily treatment with tablets containing 66 mg ferrous iron, 58 were treated with placebo. Placebo-treated: Ferritin displayed positive correlations with transferrin saturation during 2nd and 3rd trimester. There were positive correlations between ferritin, erythrocyte MCV and MCH during 2nd and 3rd trimester and postpartum. Prior to delivery and postpartum, ferritin demonstrated positive correlations with haemoglobin. Transferrin saturation showed positive correlations with MCV, MCH and MCHC during 2nd and 3rd trimester and postpartum. Transferrin saturation displayed positive correlations with haemoglobin prior to delivery and postpartum. Iron-treated: In general, there were no correlations between iron status markers. Positive correlations appeared postpartum between ferritin, transferrin saturation and MCHC but not with haemoglobin. Transferrin saturation showed a positive correlation with MCH postpartum, but not with haemoglobin. Conclusion: The patterns of relationships in placebo-treated women were consistent with iron deficient erythropoiesis. PMID- 11399597 TI - Erythropoiesis: Current Clinical Practice: Advances in the Genetics and Biology of Fanconi Anaemia. AB - The autosomal recessive disorder Fanconi anaemia (FA) has been the subject of intense study for over a decade. The genes mutated in FA patients are being cloned, but so far, the sequences of these genes have not given any clear indication of their function. Various models for the function of the FA proteins have been postulated to explain the spontaneous chromosomal abnormalities and clastogen sensitivity described in FA cells. This review summarises the critical experimental evidence for and against these models, and attempts to give some indication of the possible mechanisms by which mutations in FA genes cause patients to suffer pancytopaenia and acute myeloid leukaemia, as well as an increased risk of other malignancies. PMID- 11399598 TI - Malignancy: Tumor Suppressor Gene Aberrations in Acute Myelogenous Leukemia. AB - Acute myelogenous leukemia is a heterogeneous disease that appears to evade the normal regulatory controls of tumor suppressor genes. Studies in AML have documented mutations in both p53 and Retinoblastoma (Rb) genes, but these mutations are relatively uncommon, especially compared to their mutational frequency in solid tumors. In addition, expression abnormalities have now been documented in several tumor suppressor genes or related genes including MDM2, p73, Rb, p14(ARF), p15(INK4B), and p16(INK4A). We review the current literature regarding tumor suppressor genes in AML and suggest how these genes may be involved in the development of the disease. PMID- 11399599 TI - Malignancy: Myeloma - the Elusive Cure. AB - Myeloma accounts for 1% of all malignancies and is characterised by the malignant expansion of plasma cells. It is essentially incurable with a median survival of 6 months untreated. Typically disseminated at presentation, myeloma requires systemic therapy, in those patients with advanced or progressive disease. Of single agent therapies, oral mephalan remains the gold standard, either alone or in combination with steroids. 40% will respond with an increase in median survival from under 1 year to around 36 months. Combination chemotherapy produces an advantage in remission induction but no clear benefit for either survival or remission duration has been shown over standard melphalan and prednisolone. Significant melphalan dose escalation produces increased response rates and increased complete remission rates, however at the expense of increased toxicity. This has been ameliorated to an extent using stem cell support, however, although autografting may be superior to conventional chemotherapy it is not a curative treatment and modifications such as purging or tandem procedures remain to be fully evaluated. Allogeneic transplantation is only an option in a tiny minority of patients and has a high treatment-related mortality. Interferon alpha may improve the relapse free survival but has little effect on overall survival as the majority of patients will relapse. Experimental approaches, such as P-gp inhibition or thalidomide, are appropriate in this poor prognosis group. In many patients palliative care remains the only therapeutic option and close attention should be therefore paid to the prevention and treatment of renal failure, anaemia, infection, hypercalcaemia and bone disease. PMID- 11399600 TI - Malignancy: Identification of Predictors of Disease Status and Progression in Patients with Myeloma (MM). AB - Multiple studies have attempted to recognize the best markers of disease activity and outcome in myeloma (MM). Our objective was to identify the best variables that can reflect MM disease status. Design and methods: The data obtained from all the following tests were included in the analysis: serum levels of the 2 growth factors known to be crucial for MM growth (i.e. IL-6, and sIL-6R), routine peripheral blood data (Hb%, serum calcium, albumin, CRP, B2m, LDH) and bone marrow plasma cell (BMPC)%, as well as the age and sex of patients. The study was conducted on 21 cases of MM under chemotherapy (aged 48-74 years; M/F = 13/8) and 12 matched normal individuals. The patients were categorized into 2 groups according to their clinical status: Group#1 (n = 16; cases in plateau/stable phase), and Group#2 (n = 5; advanced/refractory cases). Results: Student t-test confirms that serum IL-6 and sIL-6R are the most statistically different variables upon comparing cases in plateau phase (Group#1) with those of advanced disease (Group#2). Stepwise discriminant analysis of data has resulted in a function that is composed of the 2 most salient variables (i.e. serum IL-6, sIL 6R). The proposed function was highly significant (p = 0.0000) with Wilk's Lambda = 0.02538. The diagnostic capability of the proposed function was very high (percent of grouped cases that were classified correctly= 100%). Conclusion: measurement of serum IL-6 and sIL-6R gives the best prediction of disease activity in patients with MM. PMID- 11399601 TI - Malignancy: Meningeal Myeloma: A Case Report and Review of Literature. AB - Purpose: Report and review of an uncommon complication of multiple myeloma. Methods: We report a case of leptomeningeal myeloma and using Medline, review previously reported cases. Results: We found 35 previously reported cases of meningeal myeloma. Of the 36 cases, including the present case, the male to female ratio was 2:1 with a median age of 58 years (range, 31 to 83 years). Forty five percent of the patients had circulating plasma cells and 90.9% had plasma cells in the cerebrospinal fluid. All but 4 cases were diagnosed antemortem. Thirty patients had a pre-existing diagnosis of multiple myeloma, while leptomeningeal disease was the presenting feature in six. Management included a combination of intrathecal chemotherapy and radiation therapy, radiation therapy alone or intrathecal chemotherapy alone in 14, 3, and 3 patients respectively. Thirteen patients also received systemic chemotherapy in addition to some form of local therapy. The median survival for the entire group was 8 weeks (range 4 days to 18 months). Even those who initially responded to local therapy relapsed in the central nervous system (CNS) or succumbed to systemic disease. Five of six patients in whom CNS disease was the presenting feature survived 4 weeks or less. Conclusions: Leptomeningeal involvement is a rare and fatal complication of multiple myeloma with a short survival despite aggressive intrathecal and systemic chemotherapy and radiation therapy. PMID- 11399602 TI - BMT: Bone Marrow Transplant Associated Thrombotic Microangiopathy. AB - Thrombotic microangiopathy is a severe microvascular disorder which may occur in up to 70% of patients undergoing bone marrow transplant. Clinically the term thrombotic microangiopathy encompasses a wide spectrum of syndromes, most importantly the thrombotic thrombocytopenic purpura (TTP) and hemolytic uremic syndrome (HUS). Thrombotic microangiopathy is characterized by the presence of thrombocytopenia, microangiopathic hemolytic anemia, renal impairment, neurological disturbances and multiorgan failure. Several causative agents have been advocated as triggering factors for bone marrow transplant associated thrombotic microangiopathy, including cyclosporine, FK506, the use of total body irradiation, infections and the presence of severe graft-versus-host disease. Plasma exchange represents the standard treatment for patients who develop TTP/HUS after bone marrow transplant, however, the mortality rate still remains high despite aggressive therapy. PMID- 11399603 TI - Erythrocytes: Anemias in Chronic Liver Diseases. AB - Anemia is a frequently observed manifestation during the clinical course of chronic liver disease. In this study, we retrospectively reviewed the hospital files of 500 chronic liver disease patients and assessed the frequency, etiology and morphology of anemia in 50 patients who fulfilled the criteria to be included in the study. The mean age of the patients was 48+/-16 years and male/female ratio was 1.4/1. The mean hemoglobin value was 9.54+/-2.03 g/dl. The mean MCV was 82.9+/-10.52 fl. Iron deficiency anemia, defined as absent bone marrow iron stores, was the most common anemia present in 50% of patients. Classical laboratory criteria used in the diagnosis of iron deficiency anemia (MCV < 80 fl, ferritin < 10 ng/ml) could not be applicable to all of the patients with iron deficiency anemia and hepatic disorders. Hemolytic anemia due to hypersplenism was the second most common anemia (24%) followed by anemias, namely anemia due to gastrointestinal hemorrhage (22%), anemia of chronic disease (8%), beta thalassemia major (8%), folate deficiency (6%), vitamin B12 deficiency (4%), macrocytic anemia (2%), aplastic anemia (2%) and immune hemolytic anemia (2%). Twenty-eight percent of the patients had more than a single cause of anemia. Morphologically, microcytic anemia was the most common seen in 46% of the patients followed by normocytic (42%) and macrocytic anemia (12%). As patients do not always present with classical laboratory findings and may have more than a cause of anemia, a complex diagnostic approach should be considered in anemic patients with hepatic disorders. PMID- 11399604 TI - Erythrocytes: Better Tolerance of Iron Polymaltose Complex Compared with Ferrous Sulphate in the Treatment of Anaemia. AB - BackgroundAbsolute iron deficiency, irrespective of aetiology, remains a major and worldwide cause of morbidity. After correction of the causative lesion, reconstitution of haemoglobin level and body iron stores is traditionally achieved with oral administration of ferrous salts. The latter have significant gastrointestinal tract side-effects that, in the short-term, may impair compliance. With protracted administration these products can cause lipid peroxidation which, in turn, may accelerate atherogenesis. An alternative formulation is an iron polymaltose complex where animal data supported a promoting effect of glycerophosphate. Setting and Trial Design This was a single centre, open, randomised, multidose four treatment parallel group study. A standard dose of ferric polymaltose complex with two differing levels of glycerophosphate was compared with an equivalent amount of iron supplied as ferrous sulphate in anaemic volunteer blood donors. The endpoints were rate of haemoglobin rise and re-expansion of body iron stores reflected in blood ferritin concentration, as well as percentage saturation of transferrin. Secondary observations were changes in the proportion of hypochromic red cells during the course of treatment, erythropoietin levels and tolerability of the two formulations. Results Outcome in the rat model suggested that the utilisation of iron from polymaltose might be enhanced by glycerophosphate. However, in donors this difference was not evident and, accordingly, the data from the three polymaltose groups combined and compared to those receiving ferrous sulphate. The rate at which haemoglobin level improved, red cell indices returned to normal, and the number of hypochromic and microcytic red cells fell was not significantly different between the groups. Similarly the serum iron, percentage saturation of transferrin and red cell ferritin were comparable. In contrast the serum ferritin levels were higher for those receiving ferrous sulphate. Additionally, side effects were significantly more frequently encountered with the latter preparation. Conclusion These data demonstrate that the addition of glycerophosphate, observed to be beneficial in rats, did not occur in humans. Secondly, in the blood donors, equivalent amounts of iron provided as the polymaltose, with or without glycerophosphate or ferrous sulphate, corrected haemoglobin concentration and morphologically abnormal erythropoiesis at comparable rates. Similarly iron stores are replenished to an equivalent extent as seen in the matching percentage saturation of transferrin and red cell ferritin levels. Interestingly, there is a discrepancy in the serum ferritin which is higher with the salt and this may reflect oxidative stress. Thirdly, corresponding efficacy can be achieved with better patient tolerance for the complex. Finally it is postulated that the iron polymaltose complex formulation more closely approximates the way in which enterocytes handle dietary iron and thus physiologic regulatory mechanisms would be expected to reciprocally slow down absorption as stores expand. Logically, therefore and, if confirmed, the latter finding suggests that this formulation may have a potential role in longer term supplementation programmes. PMID- 11399605 TI - Bone Marrow TransplantationCurrent Clinical Practice: High-Dose Chemotherapy and Stem Cell Transplantation for Non-Hodgkin's Lymphoma: Review of Recent Advances. AB - The past decade has seen impressive achievements in the development of HDT/SCT for NHL, but much remains to be accomplished. Attention can be focused now on high risk patients whose outcome with HDT/SCT, as currently practiced, is poor. This is particularly true for patients with refractory or resistant disease. The preliminary research work summarized in this review leads us to believe that further progress is forthcoming, to the benefit of the patient's survival and quality of life. PMID- 11399606 TI - Malignancy: Gene Therapy Vaccines in Acute Myeloid Leukemia : A Need for Clinical Evaluation. AB - In the last decade our understanding of the processes that govern cell growth and differentiation, malignant transformation, and metastasis has become quite sophisticated. These new insights have revolutionized our ability to diagnose and to formulate prognoses for patients with cancer, and have inspired the design and development of novel therapeutic strategies that are based on modern gene transfer technologies and act at the gene level. Gene therapy, broadly defined as the introduction of genetic material (transgenes) into a patient's cells with an intent to confer a therapeutic benefit, represents the most direct application of recombinant DNA technology in the clinical setting. The challenging concept of modifying the genetic properties of human cells captivated very quickly the interest of clinical and molecular oncologists, and currently, numerous gene therapy clinical trials in cancer patients are under investigation worldwide. Most of these studies involve manipulating the patient's immune response to tumors. The identification of tumor-specific antigens stimulating humoral and cellular responses in cancer patients, together with a better understanding of the molecular mechanisms controlling T cell activation have dramatically accelerated the search for potent cancer vaccines. In this review, we highlight important principles of cancer immunity and cancer vaccines, we discuss critical features of genetic manipulation of tumor cells, and particularly focus on preclinical studies on gene therapy vaccines in acute myeloid leukemia (AML). PMID- 11399607 TI - Malignancy: Changes in Circulating Immature and Mature Dendritic Cells During IL 2 Cancer Immunotherapy and Their Relation with Lymphocyte Increase and Clinical Response. AB - Lymphocytosis is the main biomarker predicting the efficacy of subcutaneous IL-2 anticancer immunotherapy. In addition, it has been demonstrated the fundamental role of dendritic cells (DC) in the generation of an effective anticancer immunity. However, the relation between IL-2 and DC system needs to be further understood. This preliminary study was performed in an attempt to analyze changes in circulating DC during IL-2 cancer immunotherapy in relation to lymphocyte variations and clinical efficacy of treatment. The study included 20 metastatic renal cell cancer patients, who underwent subcutaneous low-dose IL-2 immunotherapy (6.000.000 IU/day for 6 days/week for 4 weeks). To evaluate DC, venous blood samples were collected before and after 2 weeks of IL-2 injections, corresponding to the period of maximum lymphocytosis. Immature (CD123(+) ) and mature (CD11c(+) ) DC were measured by FACS and monoclonal antibodies. IL-2 induced a significant increase in the mean number of circulating mature DC, whereas no substantial change occurred in immature DC mean number. The increase in mature DC was associated with a control of disease, whereas no rise was observed in patients who had progressed on IL-2 immunotherapy. Moreover, the increase in mature DC mean number was significantly higher in patients showing evident lymphocytosis, with lymphocyte enhancement greater than 1000 cells/mmc, than in patients with less pronounced lymphocytosis, even though no significant correlation was seen in between mature DC and lymphocyte increase. This preliminary study would suggest that IL-2 may stimulate DC system and that the clinical anticancer efficacy of IL-2 is associated with the increase in circulating mature DC, which could be considered as a new favourable biomarker during IL-2 immunotherapy. PMID- 11399608 TI - MalignancyCase Report: Systemic Sarcoidosis Preceeding Acute Myeloid Leukemia. AB - The authors report the case of a 59-year-old woman who developed acute myeloid leukemia nine months after the resolution of systemic sarcoidosis treated by corticosteroid therapy. This rare case poses the questions of the physiopathogenic mechanisms, particularly a granulomatous reaction to leukemic antigens or to cytokines or of the role of an infectious agent. PMID- 11399609 TI - Hematopoiesis: Gap Junction Intercellular Communication is Likely to be Involved in Regulation of Stroma-dependent Proliferation of Hemopoietic Stem Cells. AB - The 80-100 fold increased immunohistological expression of the Gap Junction (GJ) protein Connexin-43 in murine bone marrow during the neonatal period and directly following cytoreductive treatment of adult mice suggests that the regulation of stem cell proliferation may involve GJ Intercellular Communication (GJIC). Using a series of stromal cell lines from foetal liver and neonatal bone marrow we observed that the percentage of cells with GJIC, as indicated by dye-coupling using microinjection of lucifer yellow, correlated with the stromal support for late appearing clones formed by primitive stem cells (CAFC week 3-5). In order to functionally block all GJIC between mutual stromal cells and stromal cells and hemopoietic cells, in long-term stroma-supported flask (LTC) and CAFC cultures, the lipophilic compounds amphotericin-B (AB), nystatin, alpha-glycyrrhetinic acid, tetraphenylboron, dipicrylamine and arachidonic acid were tested for their effect on GJIC and CAFC support. Only AB and nystatin, which induced complete and prolonged GJIC blockade, were able to dramatically inhibit cobblestone area (CA) formation and CFU-C generation in LTC. This inhibition could be fully abrogated by withdrawing AB within the first 2 weeks of culture. Low AB concentrations stimulated CA formation. The AB-mediated inhibition of hemopoiesis probably involved direct stromal contact with stem cells because a) AB did not inhibit CFU C generation when stem cells were cultured in trans-well inserts above the stroma; b) conditioned media from AB-containing or normal LTC did not inhibit colony formation by normal cells in semi-solid, non-stromal cultures, and c) AB did not inhibit colony formation by bone marrow cells in semi-solid culture nor did it inhibit growth or maintenance of stromal cells. In addition, The inhibition of hemopoiesis by AB could also not be explained by changes in the amount of cytokine and chemokine transcripts, including TGF-b1, in AB-blocked stromal cells. Our findings support the involvement of GJIC in stroma-dependent regulation of hemopoietic stem cell proliferation. PMID- 11399610 TI - Hematopoiesis: The Role of Stromal Integrin Interactions in Pro-B Cell Proliferation. AB - Recent studies using long term bone marrow cultures have concluded that adherence of lymphoid precursors to the underlying stromal cells is required for normal B cell development. However, the function of specific integrin interactions in B cell development remains unresolved. In our laboratory, we observed that maximal proliferation of pro-B cells required the presence of stromal cells and that stromal cytokines alone could not replace the requirement for stromal cell contact. For that reason, we questioned whether integrin interactions play a role in regulating pro-B cell proliferation in the bone marrow. Murine pro-B cell line Cl.92 expressed VLA-4, CD44, and fibronectin-receptor. Abrogation of binding of these molecules to stromal cell ligands using blocking antibodies resulted in failure of pro-B cell adherence and significant decreases in pro-B cell proliferation. Disruption of single integrin interactions did not compromise either adhesion of pro-B cells to stromal cells or IL-7 stimulated proliferation. Taken together, these data suggest that normal pro-B cells interact with stromal cells through multiple integrin interactions and that integrin mediated potentiation of pro-B cell proliferation is functionally redundant and not affected by failure of single ligand interactions. PMID- 11399611 TI - Plasma Proteins Case Report: Massive Bone Marrow Necrosis with Polyclonal Hypergammaglobulinemia and Blue Toe Syndrome. AB - Massive bone marrow necrosis was rare, and most of these cases were accompanied with malignant disease. We report a case that was thought to be idiopathic massive bone marrow necrosis. It was a 58 y.o. male who was admitted because of blue toe syndrome and hypergammaglobulinemia. We tried to detect malignant diseases with computed tomography and gallium scintigraphy, and infectious diseases with bacterial culture and viral antibodies, but all of them were negative. Pancytopenia and bone marrow necrosis was not improved, and he had died after 5-month hospitalization. Autopsy revealed massive bone marrow necrosis and bone marrow fibrosis after necrosis, but malignant or infectious diseases were not detected. It may be diagnosed as idiopathic massive bone marrow necrosis. PMID- 11399612 TI - Therapy: Hyperbaric Oxygen as the Only Effective Treatment in Mutilating and Resistant Systemic Vasculitis. AB - A forty year old man was seen in 1984 with a four year history of a painful vasculitis that responded transiently to plasma exchange. Diagnosis was revised to atypical pyoderma gangrenosum with further temporary benefit from lamprene and continuing maximally tolerated corticosteroids. The course fluctuated over the next ten years with gradual and increasing soft-tissue damage coupled with superimposed skin infections. A variety of organisms were isolated from the ulcerated areas, with each episode successfully managed on the basis of local debridement and appropriate antibiotic administration. In 1995, with extending skin devascularization, infectious bacterial episodes became more frequent and deep non-healing ulcers led to constant pain with virtual incapacity. In response to protocol hyperbaric oxygen therapy there was immediate reversal of the cutaneous damage, granulation tissue formed and new skin grew to cover the previous extensive deficits. As the lesions in his hands and feet improved so did his quality of life, with the patient again becoming ambulant and returning to work. Vascular access had become a major problem, and venography showed extensive occlusion with collateral circulation. A standard Hickman catheter was placed through the femoral vein into the inferior vena cava and functioned well over the next five years. At the end of 1996 the patient was admitted with an acute chest pain that was complicated by a major pulmonary embolus, from which he could not be resuscitated. This anecdotal experience demonstrates the important but underutilised benefits of hyperbaric oxygen in managing refractory, painful and penetrating skin ulcers. The cost of obtaining wound healing with reduction in pain by this form of treatment was approximately one-fifth of expenditure on previously ineffective management. PMID- 11399613 TI - Acute Myeloblastic Leukemia: Management with High-Dose Cytosine Arabinoside, Daunorubicin and Marrow Transplantation; Malignancy; Current Clinical Practice. AB - Combination high-dose cytosine arabinoside (ARA-C) and daunorubicin (DNR) for primary remission induction of patients with acute myeloblastic leukemia (AML) was evaluated in a single institution study. Patients aged 55 or less with an HLA sibling received an allogeneic bone marrow transplant (alloBMT) in first remission; other responders were offered autologous BMT (autoBMT). For remission induction 93 patients aged less than 60 received DNR 45 mg/m(2) BSA x 3 and ARA-C 2 gm/m(2) BSA every 12 hours for 12 doses; 53 aged 60 or older DNR 25 mg/m(2) daily x 3 and ARA-C 1.5-2.0 gm/m(2) BSA every 12 hours for 12 doses. Consolidation doses of DNR were the same but ARA-C 100 mg/m(2) BSA/day x 5 was given by continuous intravenous infusion. The complete remission rate for patients less than 60 years was 69.9% (95% CI: 59.5-79.0%) and 47.2% (95% CI: 33.3-61.4%) for the older patients. The median duration of first remission for the younger patients was 13.0 months and of overall survival 17.9 months; for patients over 60 years 5.6 and 10.0 months respectively. Disease-free survival and overall survival of the 19 patients receiving alloBMT and the 13 patients undergoing autoBMT aged less than 55 years and in first or second complete remission were significantly increased compared with 22 patients in remission but not having BMT (p < 0.001 and p < 0.013). The results support the effectiveness of high-dose ARA-C for remission induction, a need for intensive consolidation therapy and a role for BMT in the management of AML. PMID- 11399614 TI - Current Perspectives on the Use of Growth Factors in the Therapy of Acute Myeloid Leukaemia; Malignancy. AB - Recombinant haematopoietic growth factors have been available for clinical use for over a decade, however their role in the management of patients with acute myeloid leukaemia (AML) has yet to be established. There are several potential roles for the use of growth factors in the management of patients with AML, including reduction in the infective complications associated with the underlying disease and its treatment, use as mobilising agents in stem cell transplantation and as priming agents with chemotherapy. Clinical trials have failed to give clear indications for the use of growth factors following chemotherapy, mainly due to the variability of patient populations, chemotherapy and growth factor schedules used. G-CSF appears to be associated with no negative impact on remission rate or survival but clear benefits in terms of infection-related endpoints were not universally seen. Three studies did show a reduction in duration of hospitalisation, particularly when G-CSF was used following consolidation chemotherapy and economic analyses have also shown financial advantages to the administration of G-CSF. GM-CSF had a variable impact on survival and only two studies demonstrated reduction in serious infections or antimicrobial therapy use. These trials also showed economic benefits for the use of GM-CSF. Clinical studies which have attempted to exploit possible potentiation of chemotherapeutic activity by recruitment of leukaemic cells into the cell cycle have generally been disappointing. Use of growth factors for this purpose, outside the context of randomised clinical trials cannot be recommended. GM-CSF may have a role in modulating the cellular immune response against cancer cells but experimental data on its activity against leukaemia cells is limited. Augmentation of white cell function by G-CSF or GM-CSF may also be of clinical benefit in patients with suspected or confirmed fungal infection and further trials are underway. PMID- 11399615 TI - Infusional 5-fluorouracil in Cutaneous Leukaemia: a Novel Palliative Therapy in Haematological Malignancy; Malignancy. AB - We describe the management of a woman suffering from acute myelomonocytic leukaemia with cutaneous involvement. Following treatment with conventional chemotherapy she was in complete remission, but then suffered an isolated skin relapse. In order to limit side effects and myelotoxicity, an experimental therapy using a continuous infusion of 5-fluorouracil was employed. This was completely successful in eradicating the skin lesions, the patient suffered no side effects and blood counts remained within normal limits throughout. Infusional 5FU may have a role in palliative therapy in haematological malignancy. PMID- 11399616 TI - Late Effects of Hematopoietic Stem Cell Transplantation; Bone Marrow Transplantation. PMID- 11399617 TI - G-CSF Primed Autologous Marrow Harvest and Transplantation in Cytapheresis "Mobilization Failure" Patients: A Descriptive Analysis; Bone Marrow Transplantation. AB - Fifteen cancer patients, deemed blood HSC "mobilization failures" due to CD34 + cell yields of < 0.5 x 10(6) /kg from two consecutive daily cytaphereses, underwent G-CSF primed autologous bone marrow harvest in an attempt to obtain adequate hematopoietic support for subsequent autotransplantation. CD34 + cell yields from the primed marrow harvest were variable; however, some patients had > 5-fold increases in CD34 + cell yields in the marrow compared to cytapheresis, and 4 patients had CD34 + cell yields of > 1.0 (i.e., 1.2, 1.44, 1.61 and 2.45) x 10(6) /kg from the primed marrow harvest. None of the five patients previously exposed to stem cell toxins or fludarabine achieved > 0.85 x 10(6) /kg CD34 + cells with the primed marrow harvest. A significant difference was noted between G-CSF primed blood and marrow for CD34 + cells but not for GM-CFU ( p = 0.011 and p = 0.135, respectively, paired t-test). All evaluable patients engrafted; a median ANC > 0.5 x 10(9) /L recovery was achieved on D + 12 (range + 9 to + 17) in 12 of 13 evaluable patients - one died on D + 9 without recovery. The last day of platelet transfusion occurred at a median D + 13 (range + 8 to > + 66); only one patient remained platelet transfusion-dependent beyond D + 34. As anticipated, patients with higher numbers of CD34 + cells transplanted had somewhat more rapid recoveries. Although stem cell damage is obviously a key factor in mobilization failure patients, these findings raise the possibility that poor mobilization, at least in some patients, results from a mechanism other than, or in addition to, simple stem cell damage. Moreover, they raise the issue of the minimum number of marrow CD34 + - or more arguably other - cells needed for adequate short- and long-term reconstitution. The role of G-CSF in this situation, especially regarding dose and/or schedule, is intriguing but remains to be clarified. G-CSF primed marrow harvest is a potential option in certain poor mobilizers but, as fully expected, is frequently inadequate. Whether such is preferable to "steady-state" marrow harvest, continued or repeated G-CSF primed cytapheresis (with or without chemotherapy), or primed marrow with G-CSF in other schedules - or with other cytokines - is unclear and will be the subject of further study. PMID- 11399618 TI - Megakaryocytic Maturation is Regulated by Maintaining a Balance Against Cytokine Induced-cell Proliferation: Steel Factor Retards Thrombopoietin-induced Megakaryocytic Differentiation While Synergistically Stimulating Mitogenesis; Hematopoiesis. AB - Using a factor-dependent cell line MO7ER, which contains a stably transduced human erythropoietin (EPO) receptor gene in human megakaryoblastic cell line MO7e and which resulted in concomitant expression of EPO receptor, c-Mpl and c-Kit, we investigated the biological effects of these cytokines in terms of cell growth and differentiation. Thrombopoietin (TPO), EPO and Steel factor (SLF) all stimulated MO7ER cell proliferation in a dose-dependent manner. Combined stimulation of cells with SLF plus either TPO or EPO resulted in striking synergistic enhancement of MO7ER cell growth as compared with each cytokine alone, whereas combination of TPO plus EPO showed only an additive effect on cell proliferation. With regards to cell differentiation, either TPO or EPO treatment induced enhancement of platelet glycoprotein (GP) IIb/IIIa and GPIb expression. SLF induced GPIIb/IIIa and GPIb expression, but the effect was much weaker than that of EPO or TPO. However, addition of SLF to either TPO- or EPO- containing cultures (which induced potent mitogenesis in MO7ER cells) resulted in suppression of these megakaryocyte specific antigens. Addition of low-dose cytosine arabinoside (Ara-C)(1 to 10 ng/ml) enhanced TPO- or EPO- induced megakaryocytic differentiation in MO7ER cells while mildly suppressing cell growth. Treatment the cells with low-dose Ara-C plus TPO plus SLF overrode the proliferative enhancing effects of SLF and induced GPIIb/IIIa and GPIb expression as efficient as TPO alone. Retardation of TPO-induced megakaryocytic maturation was also observed in normal murine bone marrow cells by combined stimulation with TPO and SLF as assessed by the numbers of acetylcholinesterase staining-positive cells and megakaryocyte nuclear polyploidy. These results suggest that megakaryocytic maturation is, at least in part, regulated by countering cytokine induced cell proliferation. PMID- 11399619 TI - Sequential Variations in the Content of Bone Marrow Colony-forming Cells in Individual Patients with Aplastic Anemia Before and After Immunosuppressive Therapy; Hematopoiesis. AB - Previous studies have shown that the levels of hematopoietic progenitor cells (colony-forming cells; CFC) are drastically reduced in the vast majority of patients with aplastic anemia (AA). This has been observed both in patients before and after immunosuppressive therapy. In those studies, however, both groups of patients were usually formed by different individuals, thus it was not possible to follow the kinetics of such cells in each particular patient. In the present study, we have determined the content of myeloid and erythroid CFC in individual AA patients before and after therapy. Treated patients were studied at two different times (8-18 months apart) to detect any possible variations due to the ongoing treatment. At diagnosis, the levels of both myeloid and erythroid CFC were drastically reduced, as compared to normal bone marrow, in all the patients studied. This correlated with very low levels of leukocytes and hemoglobin in circulation. After the patients entered an immunosuppressive treatment, all of them showed significant increments in their CFC levels, and this correlated with increments in their hematological parameters in peripheral blood. However, in most patients CFC levels were still below the normal range. When the second sample after treatment was obtained, great variations in CFC numbers were observed. In terms of erythroid CFC levels, a further increase was seen in most patients, and this correlated with a further increase in hemoglobin levels. In contrasts, the levels of myeloid CFC were increased in only some of the patients, whereas in others, significant reductions were evident. Interestingly, in this latter group of patients, CFC never reached the levels observed before treatment. Our results indicate that, in a significant proportion of patients, a common pattern seems to exist. That is to say, low CFC numbers are present before treatment; an increase in the numbers of such cells results as an effect of the immunosuppressive therapy and further variations in CFC numbers (within individual limits that may differ significantly from one patient to another) take place as long as the treatment continues. Finally, we observed a correlation between CFC levels and the clinical status of the patients, i.e., those patients that showed a complete or a partial response to treatment showed higher levels of both myeloid and erythroid CFC than those patients that did not respond to therapy. PMID- 11399620 TI - Relationship Between Gestational Iron Deficiency and Iron Deficiency in the Newborn; Erythrocytes. AB - Objective: To investigate the iron status of women at term and its impact in their neonates. Study design: Serum ferritin (SF) was measured in 201 women at term and in their newborns. The relationship between maternal and infant SF and hemoglobin (Hb) and the effects of partial prenatal care and iron supplementation were analyzed. Results: 86% of the women had iron deficiency (ID) at term and 46% were anemic. 13 (7.5%) of the babies born to the 172 iron deficient mothers were also iron deficient, but none of the babies born to the 29 mothers with SF levels > 12 &mgr;g/L. The mean cord ferritin level (103.6 +/- 75 &mgr;g/L) and Hb (164 +/- 20 g/L) were significantly higher in babies born to mothers who had SF > 12 &mgr;g/L, compared to babies born to iron deficient mothers (73.5 +/- 49 &mgr;g/L and 156 +/- 16 g/L). The respective values in iron deficient babies were 7.1 +/- 3.5 &mgr;g/L and 157 +/- 9.0 g/L, compared to 82.8 +/- 52.8 &mgr;g/L and 162 +/- 17.5 g/L in the iron sufficient babies. Conclusion: ID was diagnosed in 86% of women at term and in 7.5% of their neonates. A placental iron threshold, limiting iron acquiscition by the fetuses of women with severe ID, is suggested. PMID- 11399621 TI - Stem Cell Transplantation for Chronic Lymphocytic Leukaemia: Is this the Way Forward in the New Millennium?; Malignancy; Current Clinical Practice. AB - The failure of conventional chemotherapy to eradicate chronic lymphocytic leukaemia cells and induce cure has led many clinicians to investigate the use of high dose chemotherapy and haemopoietic stem cell rescue in this disease. The selection of patients remains a major problem because this is a disease of elderly patients with a median overall survival of 7 years and will only therefore be applicable to a minority of patients. However transplantation is the most likely therapeutic option at this time to lead to cure in this condition. The best type of transplant is not known and not all patients will be able to mobilise adequate numbers of stem cells or have a suitable donor identified. Autologous transplantation relies on the ability of high doses of chemotherapy to eradicate disease whilst allogeneic transplantation attempts to harvest the graft versus leukaemia effect that has been identified in chronic granulocytic leukaemia. However, the high treatment related mortality and morbidity of allogeneic transplants has led to interest in the use of non-myeloablative allografts which hope to maximise the immunological effects of transplantation to achieve durable remissions. To date there have been no randomised clinical trials to compare the efficacy of combination chemotherapy, autologous or allogeneic transplants and this is unlikely to happen in the near future. Other issues that need to be addressed include the timing of transplantation, the source of stem cells, the optimal conditioning regimen and the role of immunomodulation post transplantation. This review attempts to answer some of these questions. PMID- 11399622 TI - Pentoxifylline, Ciprofloxacin and Dexamethasone Improve the Ineffective Hematopoiesis in Myelodysplastic Syndrome Patients; Malignancy. AB - Twenty-five patients with a diagnosis of myelodysplastic syndromes (MDS) were randomized to either begin therapy with pentoxifylline, ciprofloxacin and dexamethasone (PCD) immediately (10 patients) or after a 12 week observation period (control arm, 15 patients). PCD was administered with the goal of suppressing cytokine-induced excessive intramedullary apoptosis of hematopoietic cells. No marked fluctuations of blood counts were noted during the period of observation. Twenty-two patients completed at least 12 weeks of therapy: 18/22 showed some type of hematologic response, 9/18 showing an improvement in absolute neutrophil count only (p = < 0.001) and 9/18 showing multi-lineage responses. No unique category of MDS responded better, however 19/25 patients had refractory anemia (RA)/RA with ringed sideroblasts. The median time to response was 6 weeks and 3/18 responding patients maintained their responses beyond a year. We conclude that hematologic improvement in response to PCD therapy supports the validity of this unique anti-cytokine approach. Future trials should combine PCD therapy with established approaches (growth factors/chemotherapy) and also should focus on identifying more effective ways of suppressing the pro-apoptotic cytokines in MDS. PMID- 11399623 TI - Neurosarcoidosis Associated with Hodgkin's Disease; Malignancy. AB - Sarcoidosis is a granulomatous disease of undetermined origin. The association of sarcoidosis with malignant lymphoma is a rare but possibly nonfortuitous event. The authors report on the case of a 26-year-old male patient presenting with lymphnode sarcoidosis which evolved into neurosarcoidosis. The evolution was responded to corticosteroid therapy. Thirty-three months after the diagnosis of neurosarcoidosis, the patient presented with cervical Hodgkin's disease which was successfully treated by chemotherapy and radiotherapy. The pathophysiological mechanisms and possible associations are briefly discussed. PMID- 11399625 TI - Introductory Commentary on Regulation of LFA-1 Expression by CD34+ Cells and Inducible Growth Factor Production by Stroma Enable Formation of Bone Marrow Compartments, Torensma et al. PMID- 11399624 TI - Hypercalcemia after High-Dose Chemoradiotherapy for Refractory Multiple Myeloma; Subject Heading. AB - A 43-year-old man with refractory myeloma underwent allogeneic bone marrow transplantation from his HLA-matched sibling. He was conditioned with TBI (12 Gy) followed by melphalan (140 mg/m(2) ). Immediately after conditioning was initiated, he began complaining of severe lumbago, and the level of serum calcium rose from 2.25 to 3.34 mmol / l. However, the biochemical markers for tumor-lysis syndrome such as potassium, uric acid, and lactic dehydrogenase remained unchanged. Hydration with saline and pamidronate were started, but he developed acute renal failure requiring hemodialysis for 3 weeks. His plasma parathyroid hormonerelated protein (PTHrP)-NH2-terminal (3.9 pmol/l) and serum PTHrP-C terminal (125.0 pmol / l) levels markedly increased immediately after conditioning. These results suggested that the increased release of PTHrP from myeloma cells, which resulted from destruction of myeloma cells by conditioning, was the primary contributes to the occurrence of hypercalcemia. We should be aware of the occurrence of hypercalcemia when high-dose therapy is to be given to patients with refractory myeloma. PMID- 11399626 TI - Regulation of LFA-1 Expression by CD34 Positive Cells and Inducible Growth Factor Production by Stroma Enable Formation of Bone Marrow Compartments; Subject Heading. AB - Leukocyte function associated antigen 1 (LFA-1) is an adhesion molecule indispensable in immune and inflammatory reactions, but its role in hematopoiesis is poorly understood. LFA-1 is considered as a marker of late stage stem cell maturation when expressed on CD34(+) bone marrow cells. We observed that CD34(+) bone marrow cells express LFA-1, that based on LFA-1 expression several subpopulations can be distinguished, and that the level of expression appeared highly variable among different donors. Unanticipated, in time course experiments we observed that CD34(+) LFA-1(-) cells expressed LFA-1 within 24 hours upon culture. These in vitro findings support the hypothesis that once contacts with bone marrow stroma are lost, LFA-1 is upregulated by default, due to the lack of negative regulating signals from stromal cells. This might also explain the widely variable expression of LFA-1 as a result of crowding of cells in the bone marrow with subsequent loss of contact with stroma and upregulation of LFA-1, thus equipping cells with adequate adhesion receptors to migrate throughout the bone marrow. Interestingly, the expression of the LFA-1 specific activation epitope L16 on these cells is low, even after culture. This demonstrated that LFA 1 is not activated, as was confirmed by low adhesion to ICAM-1. Activation of adhesion molecules is induced by several growth factors. Indeed, we show here that an osteoblastic cell line under normal conditions does hardly produce hematopoietic growth factors but these are rapidly induced after stimulation. Such rapid induction endows the bone marrow stroma with the property to modulate the adhesive strength and enabling migration through the different environments within the stroma. Prove for such compartments within the bone marrow is provided by histological data. PMID- 11399627 TI - Concluding Annotation to Torensma et al. PMID- 11399628 TI - Commentary of Involvement of E-Cadherin in the Development of Erythroid Cells, Armeneau et al. PMID- 11399629 TI - Involvement of E-cadherin in the Development of Erythroid Cells; Subject Heading. AB - The cadherins represent a large family of structurally and functionally related cell adhesion molecules involved in morphogenesis of multicellular organisms and maintenance of solid tissues. In the hematopoietic system, however, almost nothing was known about the involvement of this family. PCR screening of RNA of human bone marrow mononuclear cells with specific primers for different classical cadherins revealed that members of this family are also expressed by bone marrow cells. Here we report that E-cadherin, which is mainly expressed by cells of epithelial origin, plays a critical role in the development of human erythrocytes. FACS analysis with human E-cadherin-specific antibodies and the use of immunoaffinity columns revealed that expression of E-cadherin is restricted to defined maturation stages of the erythropoietic cell lineage. Erythroblasts and normoblasts express E-cadherin, but mature erythrocytes do not. Lymphoid and all the other myeloid cell lineages do not express E-cadherin at any developmental stage. The differentiation of the erythroid lineage in vitro could be influenced by addition of anti-E-cadherin antibodies in a concentration dependent manner indicating a direct involvement of this cell adhesion molecule in the differentiation process. In line with these in vitro data is the finding that E cadherin is down regulated during erythroleukemia on the developing erythroid cells. Our results suggest an unanticipated function of E-cadherin in the hematopoietic system. PMID- 11399630 TI - Concluding Annotation to Armeneau et al. PMID- 11399631 TI - Serum Ferritin is a Reliable, Non-invasive Test for Iron Status in Pregnancy: Comparison of Ferritin with Other Iron Status Markers in a Longitudinal Study on Healthy Pregnant Women; Erythropoiesis. AB - Background and Aims: To assess the true positive and false positive rates of the iron status markers (serum iron, serum transferrin, transferrin saturation, haemoglobin, haematocrit, mean corpuscular volume (MCV), mean cell haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), erythrocyte count) in the diagnosis of depleted iron stores (iron depletion) during normal pregnancy and postpartum. Methods: Among 120 pregnant women, 58 were randomised to placebo treatment and 62 to iron-treatment (66 mg ferrous iron daily from 14 weeks of gestation). Iron status markers were measured every 4th week during pregnancy and 8 weeks postpartum. Iron depletion was defined by a serum ferritin concentration < 16 &mgr;g/L. The 5th percentiles for the other iron status markers in the group of iron-treated women were used as cut-off values. Calculations were made in the 2nd and 3rd trimester, praepartum and postpartum. Results: In general, the true positive rates of other iron status markers in the diagnosis of iron depletion (serum ferritin < 16 &mgr;g/L) were low ranging from 0% to 52% during pregnancy and from 9% to 64% postpartum. Transferrin saturation and MCH displayed the highest true positive rates. The false positive rates ranged from 0% to 13% during pregnancy and from 4% to 17% postpartum. Haemoglobin and MCH displayed the highest false positive rates. Conclusions: The sensitivities of the other iron status markers were too low and the false positive rates too high to be of clinical value in the diagnosis of iron depletion. Despite physiologic variations due to haemodilution, the serum ferritin concentration is currently the most reliable non-invasive marker of iron status in pregnancy and postpartum. PMID- 11399632 TI - Cephalosporin-induced Hemolytic Anemia in a Sicilian Child; Erythropoiesis. AB - A 27-month-old child developed acute hemolysis on two occasions after the administration of cephalosporin. On the first occasion, hemolysis was intravascular and was due to the formation of complexes between antibodies and the drug, which bound to red blood cells and caused severe hemolysis. On the second occasion, hemolysis was extravascular and was probably due to antibody dependent cell-mediated cytotoxicity. Marked increases in levels of CD(19) (+) and CD(57) (+) CD(8) (+) cells were detected among the subpopulations of the patient's lymphocytes but only in the level of CD(19) (+) cells from the patient's father, after incubation of a sample of whole blood with a solution of cephalosporins. These results might explain the differences between the immune response of the patient and those of other members of his family and of an unrelated control. PMID- 11399633 TI - Nitric Oxide Metabolites in Sickle Cell Anemia Patients after Oral Administration of Hydroxyurea; Hemoglobinopathy. AB - The mechanism of action of hydroxyurea (HU) in decreasing the frequency of pain crisis in sickle cell disease (SCD) has not been fully elucidated. In vitro and in vivo studies suggest that nitric oxide (NO), a potent vasodilator, may partly be responsible for the beneficial effect of HU. This study was designed to determine the effect of oral administration of HU on plasma levels of NO metabolites (NO(x) ) in sickle cell patients (SCP). The results indicate that during steady-state plasma levels of NO(x) were significantly higher in HU treated patients compared to non HU-treated patients or normal controls (p <.05). In five inpatients in mild pain plasma levels of NO(x) increased significantly after 2 h of HU administration (p <.05); however, in three inpatients in persistent pain with significantly lower baseline NO(x) there was a minimal NO(x) response to HU at 2 h (p <.01). These observations indicate that HU administration is associated with the production of NO in some SCP, but that further study of the pharmacodynamics of this effect is necessary. PMID- 11399634 TI - Application of Resistance Reversal Agents in Hematologic Malignancies; Malignancy; Current Clinical Practice. AB - The clinical application of resistance reversal drugs for patients with hematologic malignancies is reviewed. The phenomenon of multidrug resistance versus other mechanisms are discussed. The pump-like mechanisms of P glycoprotein, multidrug resistance associated protein, lung resistance protein and of other ATP binding cassette transporter proteins are reviewed briefly, as well as the important substrate drugs and pump-blocking compounds. The problems associated with resistance protein assays in clinical samples and the concept of prognostic versus therapeutic clinical relevance are described, within the context of selected hematologic malignancies. Toxicities and treatment outcomes of phase II and III trials of reversal agents in lymphoma, multiple myeloma, myelodysplastic syndromes, acute myeloid leukemia and blast phase of chronic myeloid leukemia are reviewed. Finally, current options for on-study management of relapsed or refractory hematologic malignancy patients are discussed. PMID- 11399635 TI - P-Glycoprotein Expression in Acute Myeloid Leukaemia Cells at Diagnosis: Its relationship to Daunorubicin or Idarubicin Induction Therapy and Survival; Malignancy. AB - We investigated the expression of P-glycoprotein (P-gp) in 50 adults with de novo diagnosed acute myeloid leukaemia (AML) and the relationship between presence of P-gp in leukaemic cells and efficacy, as remission induction and survival rate, of two different anthracyclines, daunorubicin (DNR) and idarubicin (IDR). We found that 30 out of 50 patients (60%) were negative (Group 1) and 20 (40%) were positive (Group 2) for P-gp expression evaluated by mean of MRK16 MoAb using a cut-off of 10% positive cells. Thirty-five out of 50 patients (70%) obtained complete remission (CR); depending on P-gp expression, the CR rate was 80% for group 1 and 45% for group 2 (p < 0.005). The median duration of overall survival was 20 months for patients in Group 1 as compared with 10 months for patients of Group 2 (p < 0.005). Regarding the anthracycline used, no significant difference in CR was observed in patients of Group 1 (75% of CR with DNR vs. 90% with IDR); Group 2 obtained 40% of CR with DNR vs. 70% with IDR (p < 0.005). The median duration of overall survival (OS) with the two regimens was comparable in Group 1, while it was significantly longer in patients of Group 2 treated with IDR compared with DNR regimen (p < 0.005). These results confirm the prognostic value of P-gp expression in AML at first appearance and we suggest that idarubicin could be a valid anthracycline drug in the treatment of AML to be evaluated as potential drug of choice in patients with primary or drug-induced multidrug resistance. PMID- 11399636 TI - The AML1/MTG8 Fusion Transcript in t(8;21) Positive AML and its Implication for the Detection of Minimal Residual Disease; Malignancy. AB - The reciprocal translocation t(8;21)(q22;q22) is one of the most frequent chromosomal aberrations in acute myeloblastic leukemia (AML). At the molecular level, this aberration rearranges the gene for the AML1-transcription factor on chromosome 21, which is essential for normal hematopoiesis, to the MTG8 gene on chromosome 8, thereby leading to a specific AML1/MTG8 fusion mRNA. This fusion gene is involved in leukemogenesis presumably by interfering with normal AML1 dependent transcriptional regulation. AML patients with t(8;21) have a favourable response to chemotherapy and a relatively good prognosis after intensive consolidation treatment with high dose AraC or autologous stem cell transplantation. RT-PCR for the specific AML1/MTG8 fusion transcripts can be used for the sensitive detection of residual leukemic cells during and after therapy. However, since a considerable proportion of these patients shows a positive PCR result even in long-term complete hematological remission, the prognostic value of qualitative PCR methods is doubtful. In contrast, quantitative PCR methods might be able to identify patients with a high risk of relapse by serial quantification of minimal residual disease (MRD). Because of its high degree of standardisation and automation, the recently developed real time PCR method can be used for the valid and reproducible detection of MRD in large prospective trials. This technology offers the potential to define the antileukemic efficiency of different treatment elements and the prognostic value of MRD in patients with t(8;21) positive AML. PMID- 11399638 TI - Editorial on "Specificity of Hematopoietic Stem and Progenitor Cell Homing to Bone Marrow: A Perspective", Cheryl Hardy. PMID- 11399637 TI - Implication of Granulocyte-Macrophage Colony Stimulating Factor (GM-CSF) and Interleukin-3 (IL-3) in Children with Acute Myeloid Leukaemia (AML); Malignancy. AB - Granulocyte-macrophage colony stimulating factor (GM-CSF) and Interleukin-3 (IL 3) are increasingly used to stimulate granulopoiesis in neutropenic patients but these are rarely used in the lights of knowledge of the endogenous CSF-levels. In this study we measured serum levels of GM-CSF and IL-3 at diagnosis and after remission in children with acute leukaemia, using an enzyme linked immuno-sorbent assay (ELISA) techniques in 14 patients with acute myeloid leukaemia (AML) and 27 patients with acute lymphoblastic leukaemia (ALL). Twelve healthy age-matched children were used as a reference group. AML patients showed a highly significant increase in serum levels of GM-CSF and IL-3 before induction of therapy (p < 0.0001) compared to the reference control group, with a highly significant decline of both GM-CSF and IL-3 (p < 0.0001) after successful remission. On the other hand, ALL patients showed no significant elevation of GM-CSF and IL-3 at diagnosis (p > 0.5), with no significant difference between preinduction and postinduction serum levels of either (p > 0.5). Since these cytokines are known to be fundamental for the growth of AML cells, we postulate that the pretreatment levels of both GM-CSF and IL-3 could play a role in the pathogenesis of AML. PMID- 11399639 TI - Specificity of Hematopoietic Stem and Progenitor Cell Homing to Bone Marrow: A Perspective; Subject Heading. AB - Little is known about the hematopoietic stem and progenitor cell membrane recognition and adhesion molecules which mediate their specific patterns of movement into and out of the marrow compartment during steady state hematopoiesis and during pathological conditions. Implicit in the cellular targeting of these cells to marrow stroma, or "homing", is a high degree of molecular specificity. Identification of homing determinants and knowledge of their function in conferring specificity to these events may provide new insight into the localization of hematopoietic stem cells within the bone marrow, directly impacting clinical stem cell transplantation. In addition, a homing protein gene/promoter complex, or a stromal counter-receptor gene, may provide a valuable target for driving expression of gene constructs in early hematopoietic cells. PMID- 11399640 TI - The Role of Inflammation and Leukocytes in the Pathogenesis of Sickle Cell Disease; Haemoglobinopathy. AB - Acute and chronic vascular occlusion underlies much of the morbidity and mortality in sickle disease. Abnormal polymerization of deoxygenated hemoglobin S (HbS) resulting in stiff, non-deformable erythrocytes is central to sickle cell pathogenesis. However, a complex interplay of many factors determines the balance between adequate blood flow and vessel obstruction. Serum markers of inflammation have provided evidence for a state of chronic inflammation in sickle cell disease (SCD). Inflammation promotes endothelial adherence to sickle erythrocytes. Studies demonstrating a beneficial effect of steroid therapy for painful episodes and acute chest syndrome provide indirect evidence for the role of inflammation in this disease. Leukocytosis, in the absence of infection, is common in SCD patients and predicts for stroke, acute chest syndrome, and overall mortality. Neutrophils and monocytes have been shown to be activated in these patients. Activated leukocytes further promote vascular inflammation and vessel damage. A reduction in leukocytes, and thus inflammation, may partially explain the beneficial effects of hydroxyurea in this disease. These data provide a strong rationale for clinical studies of therapy directed at inflammation and/or leukocytes in sickle cell disease. PMID- 11399641 TI - Physiological regulation of cyclooxygenase-2 in the kidney. AB - In adult mammalian kidney, cyclooxygenase-2 (COX-2) expression is found in a restricted subpopulation of cells. The two sites of renal COX-2 localization detected in all species to date are the macula densa (MD) and associated cortical thick ascending limb (cTALH) and medullary interstitial cells (MICs). Physiological regulation of COX-2 in these cellular compartments suggests functional roles for eicosanoid products of the enzyme. COX-2 expression increases in high-renin states (salt restriction, angiotensin-converting enzyme inhibition, renovascular hypertension), and selective COX-2 inhibitors significantly decrease plasma renin levels, renal renin activity, and mRNA expression. There is evidence for negative regulation of MD/cTALH COX-2 by angiotensin II and by glucocorticoids and mineralocorticoids. Conversely, nitric oxide generated by neuronal nitric oxide synthase is a positive modulator of COX 2 expression. Decreased extracellular chloride increases COX-2 expression in cultured cTALH, an effect mediated by increased p38 mitogen-activated protein kinase activity, and, in vivo, a sodium-deficient diet increases expression of activated p38 in MD/cTALH. In contrast to COX-2 in MD/cTALH, COX-2 expression increases in MICs in response to a high-salt diet as well as water deprivation. Studies in cultured MICs have confirmed that expression is increased in response to hypertonicity and is mediated, at least in part, by nuclear factor-kappaB activation. COX-2 inhibition leads to apoptosis of MICs in response to hypertonicity in vitro and after water deprivation in vivo. In addition, COX-2 metabolites appear to be important mediators of medullary blood flow and renal salt handling. Therefore, there is increasing evidence that COX-2 is an important physiological mediator of kidney function. PMID- 11399642 TI - Optimized suspension culture: the rotating-wall vessel. AB - Suspension culture remains a popular modality, which manipulates mechanical culture conditions to maintain the specialized features of cultured cells. The rotating-wall vessel is a suspension culture vessel optimized to produce laminar flow and minimize the mechanical stresses on cell aggregates in culture. This review summarizes the engineering principles, which allow optimal suspension culture conditions to be established, and the boundary conditions, which limit this process. We suggest that to minimize mechanical damage and optimize differentiation of cultured cells, suspension culture should be performed in a solid-body rotation Couette-flow, zero-headspace culture vessel such as the rotating-wall vessel. This provides fluid dynamic operating principles characterized by 1) solid body rotation about a horizontal axis, characterized by colocalization of cells and aggregates of different sedimentation rates, optimally reduced fluid shear and turbulence, and three-dimensional spatial freedom; and 2) oxygenation by diffusion. Optimization of suspension culture is achieved by applying three tradeoffs. First, terminal velocity should be minimized by choosing microcarrier beads and culture media as close in density as possible. Next, rotation in the rotating-wall vessel induces both Coriolis and centrifugal forces, directly dependent on terminal velocity and minimized as terminal velocity is minimized. Last, mass transport of nutrients to a cell in suspension culture depends on both terminal velocity and diffusion of nutrients. In the transduction of mechanical culture conditions into cellular effects, several lines of evidence support a role for multiple molecular mechanisms. These include effects of shear stress, changes in cell cycle and cell death pathways, and upstream regulation of secondary messengers such as protein kinase C. The discipline of suspension culture needs a systematic analysis of the relationship between mechanical culture conditions and biological effects, emphasizing cellular processes important for the industrial production of biological pharmaceuticals and devices. PMID- 11399643 TI - Recovery after relief of fetal urinary obstruction: morphological, functional and molecular aspects. AB - The effects of obstruction [urinary tract obstruction (UTO)] and relief on renal development were examined in an experimental model in the fetal lamb. Bladder outlet obstruction was performed at 60 days of gestation; relief was performed by vesicoamniotic shunting at 90 days of gestation. Studies were carried out in obstructed (OF60; n = 11), shunted (SF; n = 5), and control fetuses (CF; n = 11) at 120 days of gestation. Fetal UTO produced either hydronephrosis (64%) or dysplasia (36%); dysplasia was always associated with a reduction in the number of glomeruli [950 +/- 99 (dysplasia) vs. 1,852 +/- 249 (CF) glomeruli/section]. Obstructed fetuses had lower creatinine clearance [0.76 +/- 0.41 (OF60) vs. 0.96 +/- 0.21 (CF) ml x min(-1) x kg(-1)], higher sodium fractional excretion [17.2 +/ 20.3 (OF60) vs. 2.4 +/- 3.7% (CF)], and higher urinary concentration [80 +/- 30 (OF60) vs. 43 +/- 22 (CF) micromol/l] than controls. In SF, the number of glomeruli was increased at 120 days of gestation (1,643 +/- 106 glomeruli/section) compared with nondiverted fetuses (1,379 +/- 502 glomeruli/section), and the temporal pattern of PAX2, disrupted after obstruction, was restored. In conclusion, early fetal UTO leads to either renal hydronephrosis with normal glomerular development or dysplasia with a decreased number of glomeruli; in utero urine diversion performed before the end of nephrogenesis may allow a reversal of the glomerulogenesis arrest observed. PMID- 11399644 TI - Mycophenolate mofetil prevents salt-sensitive hypertension resulting from nitric oxide synthesis inhibition. AB - Recent studies have suggested that subtle microvascular and tubulointerstitial injury in the kidney can cause salt-sensitive hypertension. To test this hypothesis, we determined whether the mild renal disease induced by transient blockade of nitric oxide (NO) synthesis would result in salt-sensitive hypertension and whether prevention of the renal injury by coadministration of the immunosuppressive agent mycophenolate mofetil (MMF) would block the development of salt sensitivity. N(omega)-nitro-L-arginine-methyl ester (L-NAME; 70 mg/100 ml in the drinking water) was administered for 3 wk to rats with or without MMF (30 mg x kg(-1) x day(-1) by gastric gavage), followed by a 1-wk "washout" period in which the MMF was continued, which was followed in turn by placement on a high-salt (4% NaCl) diet for an additional 4 wk. Renal histology was examined at 3 and 8 wk, and blood pressure was measured serially. L-NAME treatment resulted in acute hypertension and the development of mild renal injury. During the washout period, blood pressure returned to normal, only to return to the hypertensive range on exposure of the animals to a high-salt diet. MMF treatment prevented the development of hypertension in response to a high salt diet. This correlated with the ability of MMF to inhibit specific aspects of the renal injury, including the development of segmental glomerulosclerosis, the infiltration of T cells and ANG II-positive cells, and the thickening of afferent arterioles. PMID- 11399645 TI - Apical membrane of native OMCD(i) cells has nonselective cation channels. AB - The purpose of this study was to examine cation channel activity in the apical membrane of the outer medullary collecting duct of the inner stripe (OMCD(i)) using the patch-clamp technique. In freshly isolated and lumen-opened rabbit OMCD(i), we have observed a single channel conductance of 23.3 +/- 0.6 pS (n = 17) in cell-attached (c/a) patches with high KCl in the bath and in the pipette at room temperature. Channel open probability varied among patches from 0.06 +/- 0.01 at -60 mV (n = 5) to 0.31 +/- 0.04 at 60 mV (n = 6) and consistently increased upon membrane depolarization. In inside-out (i/o) patches with symmetrical KCl solutions, the channel conductance (22.8 +/- 0.8 pS; n = 10) was similar as in the c/a configuration. Substitution of the majority of Cl- with gluconate from KCl solution in the pipette and bath did not significantly alter reversal potential (E(rev)) or the channel conductance (19.7 +/- 1.1 pS in asymmetrical potassium gluconate, n = 4; 21.4 +/- 0.5 pS in symmetrical potassium gluconate, n = 3). Experiments with 10-fold lower KCl concentration in bath solution in i/o patches shifted E(rev) to near the E(rev) of K+. The estimated permeability of K+ vs. Cl- was over 10, and the conductance was 13.4 +/- 0.1 pS (n = 3). The channel did not discriminate between K+ and Na+, as evidenced by a lack of a shift in the E(rev) with different K+ and Na+ concentration solutions in i/o patches (n = 3). The current studies demonstrate the presence of cation channels in the apical membrane of native OMCD(i) cells that could participate in K+ secretion or Na+ absorption. PMID- 11399646 TI - Renal apoptosis parallels ceramide content after prolonged ureteral obstruction in the neonatal rat. AB - Obstructive nephropathy, the primary cause of renal insufficiency in infants, is characterized by progressive renal apoptosis. Ceramide is a sphingolipid known to stimulate apoptosis in the kidney. We investigated the effects of unilateral ureteral obstruction (UUO) on endogenous renal ceramide content and apoptosis in neonatal and adult rats. Animals were subjected to UUO or sham operation on the first day of life and were studied 3-28 days later. Adult rats were similarly treated and then studied 3 or 14 days later. In additional neonatal rats, the obstruction was removed after 5 days, with study at 14 or 28 days. Renal ceramide content was measured by diacylglycerol kinase assay, and apoptosis was determined by the terminal deoxynucleotidyl transferase dUTP nick-end-labeling technique. Renal ceramide content was 50-fold higher in the 3-day neonatal compared with the adult kidney and 10-fold higher in the 7-day neonatal compared with the adult kidney, but there was no additional effect of UUO on ceramide content at these ages. However, after 14 or 28 days UUO in the neonate, renal ceramide was elevated compared with sham or intact opposite kidneys, and renal apoptosis was directly related to ceramide content (r = 0.99, P < 0.001). Moreover, renal ceramide was reduced by relief of obstruction (P < 0.05). There was less apoptosis in the obstructed kidney of the adult than the neonate, and UUO had no effect on ceramide content at 14 days in the adult. We conclude that prolonged UUO (at least 14 days duration) increases endogenous renal ceramide in the neonatal but not the adult rat. It is likely that this contributes to the prolonged renal apoptotic response of the neonatal obstructed kidney. PMID- 11399647 TI - HGF promotes adhesion of ATP-depleted renal tubular epithelial cells in a MAPK dependent manner. AB - Hepatocyte growth factor (HGF) has been shown to enhance recovery from renal tubular ischemia. We investigated the possibility that HGF improves recovery by preventing ischemia-induced loss of cell adhesion. Murine inner medullary collecting duct-3 (mIMCD-3) cells subjected to 90% ATP depletion demonstrated a 55% decrease in adhesion, an effect that was completely reversed by the addition of HGF. Assays examining release of adherent cells revealed similar results with 30 min of ATP depletion causing loss of adhesion of 25% of mIMCD-3 cells and HGF completely reversing this effect. In contrast, HGF was unable to reverse the loss of adhesion of cells exposed to 99% ATP depletion. Examination of the mitogen activated protein kinase (MAPK) signaling pathway revealed that HGF could induce extracellular signal-regulated kinase (ERK) phosphorylation in control and 90% ATP-depleted cells but not in 99% ATP-depleted cells. Inhibition of ERK activation with U0126 completely blocked the HGF-dependent reversal of ATP depleted cell adhesion. Thus ATP-depleted cells demonstrate a marked decrease in cell adhesion that is reversible by the addition of HGF. This effect of HGF requires activation of the MAPK pathway. PMID- 11399648 TI - Delayed branching of endothelial capillary-like cords in glycated collagen I is mediated by early induction of PAI-1. AB - Development of micro- and macrovascular disease in diabetes mellitus (DM) warrants a thorough investigation into the repertoire of endothelial cell (EC) responses to diabetic environmental cues. Using human umbilical vein EC (HUVEC) cultured in three-dimensional (3-D) native collagen I (NC) or glycated collagen I (GC), we observed capillary cord formation that showed a significant reduction in branching when cells were cultured in GC. To gain insight into the molecular determinants of this phenomenon, HUVEC subjected to GC vs. NC were studied using a PCR-selected subtraction approach. Nine different genes were identified as up- or downregulated in response to GC; among those, plasminogen activator inhibitor 1 (PAI-1) mRNA was found to be upregulated by GC. Western blot analysis of HUVEC cultured on GC showed an increase in PAI-1 expression. The addition of a neutralizing anti-PAI-1 antibody to HUVEC cultured in GC restored the branching pattern of formed capillary cords. In contrast, supplementation of culture medium with the constitutively active PAI-1 reproduced defective branching patterns in HUVEC cultured in NC. Ex vivo capillary sprouting in GC was unaffected in PAI-1 knockout mice but was inhibited in wild-type mice. This difference persisted in diabetic mice. In conclusion, the PCR-selected subtraction technique identified PAI-1 as one of the genes characterizing an early response of HUVEC to the diabetic-like interstitial environment modeled by GC and responsible for the defective branching of endothelial cells. We propose that an upregulation of PAI 1 is causatively linked to the defective formation of capillary networks during wound healing and eventual vascular dropout characteristic of diabetic nephropathy. PMID- 11399649 TI - Atrial natriuretic peptide attenuates ANG II-induced hypertrophy of renal tubular cells. AB - ANG II arrests LLC-PK1 cells in the G1 phase of the cell cycle and induces hypertrophy, an effect mediated by induction of p27Kip1. We studied whether atrial natriuretic peptide (ANP) may modulate ANG II-induced hypertrophy and p27Kip1 expression in tubular LLC-PK1 cells. ANP, through its fragments 3---28 and 4---27, prevented ANG II-induced cell cycle arrest. ANP inhibited >80% of ANG II-induced p27Kip1 protein expression (Western blots). ANP stimulated expression of MKP-1, a phosphatase involved in dephosphorylation of p44/42 mitogen-activated protein (MAP) kinase, up to 12 h. ANP prevented the ANG II-mediated phosphorylation peak of MAP kinase after 12 h of stimulation. 8-Bromo-cGMP mimicked all the effects of ANP. Transfection with MKP-1 antisense, but not sense, oligonucleotides abolished the modifying role of ANP on ANG II-mediated cell cycle arrest. The effect of ANP on ANG II-mediated hypertrophy of LLC-PK1 cells is regulated on the level of MAP kinase phosphorylation, a key step in the induction of p27Kip1. Although ANP and ANG II both stimulate generation of reactive oxygen species, ANP additionally induces expression of MKP-1, leading to interference with ANG II-mediated MAP kinase phosphorylation. PMID- 11399650 TI - cADP-ribose/ryanodine channel/Ca2+-release signal transduction pathway in mesangial cells. AB - Signaling via release of Ca2+ from intracellular stores is mediated by several systems, including the inositol 1,4,5-trisphosphate (IP3) and cADP-ribose (cADPR) pathway. We recently discovered a high capacity for cADPR synthesis in rat glomeruli and cultured mesangial cells (MC). We sought to determine whether 1) cADPR synthesis in MC is regulated by cytokines and hormones, 2) ryanodine receptors (RyRs) are expressed in MC, and 3) Ca2+ is released through RyRs in response to cADPR. We found that ADP-ribosyl cyclase, a CD38-like enzyme that catalyzes cADPR synthesis, is upregulated in MC by tumor necrosis factor-alpha, interleukin-1beta, and all-trans retinoic acid (atRA). [3H]ryanodine binds to microsomal fractions from MC with high affinity in a Ca2+-dependent manner; binding is enhanced by specific RyR agonists and blocked by ruthenium red and cADPR. Western blot analysis confirmed the presence of RyR in MC. Release of 45Ca2+ from MC microsomes was stimulated by cADPR; release was blocked by ruthenium red and 8-bromo-cADPR. ADPR (non-cyclic) was without effect. In MC, TNF alpha and atRA amplified the increment of cytoplasmic Ca2+ elicited by vasopressin. We conclude that MC possess elements of a novel ADP-ribosyl cyclase- >cADPR-->RyR-->Ca2+-release signaling pathway subject to regulation by proinflammatory cytokines and steroid superfamily hormones. PMID- 11399651 TI - Effects of filtration rate on the glomerular barrier and clearance of four differently shaped molecules. AB - The effect of shape on the transglomerular passage of solutes has not been hitherto systematically studied. We perfused isolated rat kidneys to determine the fractional clearances (theta) at various filtration rates for four molecules of different shapes but with similar Stokes-Einstein radii (aSE = 34-36 A). The theta for hyaluronan, bikunin, and Ficoll36 A were 66, 16, and 11%, respectively, at a glomerular filtration rate (GFR) of 0.07 ml x min(-1) x g wet wt(-1) and decreased to 46, 14, and 7%, respectively, on a fivefold increase in GFR. Under the same conditions, theta for albumin increased from 0.15 to 0.74%, and similar behavior was observed for larger Ficolls (aSE >45 A). Pore analysis showed that the "apparent neutral" solute radii of Ficoll, albumin, bikunin, and hyaluronan were 35, 64, 33, and 24 A, respectively, despite similar aSE. In addition, the properties of the glomerular filter changed with increasing GFR and hydrostatic pressure. We conclude that elongated shape, irrespective of size and charge, drastically increases the transglomerular passage of a solute, an effect that is related to its frictional ratio. PMID- 11399652 TI - Functional characterization of basolateral and luminal dopamine receptors in rabbit CCD. AB - Previous studies reported the existence of both D1- and D2-like receptors in the cortical collecting duct (CCD). However, especially with regard to natriuresis, it remains controversial. In the present study, rabbit CCD was perfused to characterize the receptor subtypes responsible for the tubular actions. Basolateral dopamine (DA) induced a dose-dependent depolarization of transepithelial voltage. Basolateral domperidone, a D2-like receptor antagonist, abolished depolarization, whereas SKF-81297, a D1-like receptor agonist, showed no significant change. In addition, bromocriptine, a D2-like receptor agonist, also caused depolarization, whereas SKF-81297, a D1-like receptor agonist, did not depolarize significantly. Moreover, RBI-257, a D4-specific antagonist, reversed the basolateral DA-induced depolarization. In contrast to the basolateral side, luminal DA caused depolarization via a D1-like receptor; however the change was less than that for basolateral DA. For further evaluation, 22Na+ flux (J(Na)) was measured to confirm the effect of DA on Na+ transport. Basolateral DA also caused a suppression of J(Na), and this reaction was abolished by domperidone. These results suggested that the basolateral D2-like receptor is mainly responsible for the natriuretic action of DA in rabbit CCD. PMID- 11399653 TI - Effect of COX-2 inhibitor NS-398 on expression of PGE2 receptor subtypes in M-1 mouse CCD cells. AB - Our present study has investigated the effect of cyclooxygenase-2 (COX-2) inhibition on prostaglandin E2 (PGE2) receptor expression in M-1 cortical collecting duct cells and measured their response to PGE2. Using a semiquantitative titration analysis method, we show that following the addition of the COX-2-specific inhibitor NS-398, E-prostanoid receptor subtype (EP3 and EP4) mRNA expression was found to increase threefold each vs. the vehicle-treated control. We also observed that EP1 but not EP2 is expressed in M-1 cells and EP2 levels are not induced by NS-398. To determine the status of the PGE2 response on exposure to NS-398, we measured cAMP levels in cells after stimulation with varying concentrations of PGE2, then pretreated the cells with 10 microM NS-398 before PGE2 exposure and found a significant rise in the stimulatory effect of PGE2 on cAMP production. Finally, Western blot analysis of the levels of the EP4 receptor protein in control vs. NS-398-treated cells revealed an induction in protein levels in these cells, correlating with the induction in EP4 mRNA. We conclude that NS-398 upregulates the expression of EP3 and EP4 mRNA in M-1 cells. Also, EP4 protein levels are increased, resulting in an increased stimulation of cAMP production by PGE2. PMID- 11399654 TI - 97- and 117-kDa forms of collecting duct urea transporter UT-A1 are due to different states of glycosylation. AB - UT-A1 is an extremely hydrophobic 929-amino acid integral membrane protein, expressed in the renal inner medullary collecting duct, with a central role in the urinary concentrating mechanism. Previous immunoblotting studies in rats have revealed that UT-A1 is present in kidney in 97- and 117-kDa monomeric forms and that the relative abundance of the two forms is altered by vasopressin treatment and other treatments that altered urinary inner medullary urea concentration. The present studies were carried out using protein chemistry techniques to determine the origin of the two forms. Peptide-directed polyclonal antibodies targeted to five sites along the polypeptide sequence from the NH2 to the COOH terminus labeled both forms, thus failing to demonstrate a significant deletion in the primary amino acid chain. The 97- and 117-kDa monomeric forms were both reduced to 88 kDa by deglycosylation with N-glycosidase F, indicating that a single polypeptide chain is glycosylated to two different extents. Studies using nonionic detergents for membrane solubilization or using homobifunctional cross linkers demonstrated that UT-A1 exists as a 206-kDa protein complex in native kidney membranes. The mobility of this complex was also increased by deglycosylation. Both the 97- and 117-kDa proteins, as well as the 206-kDa complex, were immunoprecipitated with UT-A1 antibodies. We conclude that UT-A1 is a glycoprotein and that the two monomeric forms (97 and 117 kDa) in inner medullary collecting duct are the consequence of different states of glycosylation. PMID- 11399655 TI - Evidence for endothelin involvement in the response to high salt. AB - Recent evidence suggests that endothelin-1 (ET-1), perhaps through the ET(B) receptor, may participate in blood pressure regulation through the control of sodium excretion. Mean arterial pressure (MAP) was continuously measured via telemetry implants in male Sprague-Dawley rats. After 1 wk of baseline measurements, rats were given either high (10%) or low (0.08%) NaCl in chow for the remainder of the experiment (n = 5 in each group). MAP was significantly increased in rats on a high-salt diet (115 +/- 2 mmHg) compared with rats on the low-salt diet (103 +/- 2 mmHg; P < 0.05). All rats were then treated with the ET(B) receptor antagonist A-192621 mixed with the food and adjusted daily to ensure a dose of 30 mg x kg(-1) x day(-1). ET(B) blockade produced an increase in MAP within a few hours of treatment and was significantly higher in rats on the high-salt diet over a 1-wk period (170 +/- 3 vs. 115 +/- 3 mmHg, P < 0.01). To determine whether the increase in MAP during A-192621 treatment was due to increased ET(A) receptor activation, all rats were then given the ET(A)-selective antagonist ABT-627 in the drinking water while a low-salt/high-salt diet and ET(B) blockade were continued. ABT-627 decreased MAP within a few hours in rats on either the high-salt (113 +/- 3 mmHg) or low-salt (101 +/- 3 mmHg) diet. These results support the hypothesis that endothelin, through the ET(B) receptor, participates in blood pressure regulation in the response to salt loading. PMID- 11399656 TI - Angiotensin II activates the GFAT promoter in mesangial cells. AB - Expression of glutamine:fructose-6-phosphate amidotransferase (GFAT), the rate limiting enzyme for glucose entry into the hexosamine pathway, is transcriptionally regulated. Immunohistochemical studies of human kidney biopsies demonstrate increased GFAT expression in diabetic glomeruli, but the mechanism responsible for this overexpression is unknown. Given the role of ANG II in diabetic kidney disease, we chose to study the effect of ANG II on GFAT promoter activity in mesangial cells (MC). Exposure of MC to ANG II (10(-7) M) increased GFAT promoter activity (2.5-fold), mRNA (3-fold), and protein (1.6-fold). ANG II mediated GFAT promoter activation was inhibited by the ANG II type I receptor antagonist candesartan (10(-8) M) but was unaffected by the ANG II type II receptor antagonist PD-123319 (10(-8) M). The intracellular calcium chelator 1,2 bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (10(-6) M), protein kinase C (PKC) inhibitors bisindoylmaleimide-4 (10(-6) M) and calphostin C (10(-7) M), protein tyrosine kinase (PTK) inhibitor genistein (10(-4) M), Src family kinase inhibitor PP2 (2.5 x 10(-7) M), p42/44 mitogen-activated protein kinase (MAPK) inhibitor PD-98059 (10(-5) M), and the epidermal growth factor (EGF) inhibitor AG 1478 all attenuated GFAT promoter activation by ANG II. We conclude that the GFAT promoter is activated by ANG II via the AT1 receptor. Promoter activation is calcium dependent and PKC dependent but also involves PTK signaling pathways including Src, the EGF receptor, and p42/44 MAPK. PMID- 11399657 TI - Downregulation of AQP1, -2, and -3 after ureteral obstruction is associated with a long-term urine-concentrating defect. AB - Previously, we demonstrated that 24 h of bilateral ureteral obstruction (BUO) and short-term release of BUO was associated with a decrease in the expression of aquaporin-2 (AQP2), polyuria, and a reduced urinary concentrating capacity (10). The purposes of the present study were to examine whether BUO and the long-term release of BUO (BUO-R) for 3, 14, and 30 days were associated with changes in the expression of renal AQP1, AQP2, and AQP3 and whether such changes were associated with parallel changes in urinary output and urinary concentrating capacity. Rats (n = 4-7 in each group) were kept in metabolic cages for measurements of urinary output. Kidneys were removed to determine the expression levels of AQP1, AQP2, and AQP3 by semiquantitative immunoblotting. AQP2 was downregulated after 24 h of BUO (42 +/- 3%). Downregulation of AQP2 persisted 3 (43 +/- 14%; P < 0.01) and 15 days after BUO-R (48 +/- 11%; P < 0.01) but was normalized 30 days after BUO-R. AQP3 showed a similar pattern. Moreover, AQP1 was downregulated in response to BUO (65 +/- 7%) and remained downregulated 3 days after BUO-R (41 +/- 5%), 14 days after BUO-R (57 +/- 8%), and 30 days after BUO-R (59 +/- 5%). BUO-R resulted in a significant polyuria that gradually decreased, although it remained significant at day 30. Urinary concentrating capacity remained significantly impaired when determined 3, 14, and 30 days after BUO-R in response to a 24-h period of thirst (1,712 +/- 270 vs. 2,880 +/- 91 mosmol/kgH2O at day 30, P < 0.05). In conclusion, the expression of AQP1, AQP2, and AQP3 were long-term downregulated after BUO-R, suggesting that dysregulation of aquaporins located at the proximal tubule, thin descending limb of the loop of Henle, and the collecting duct may contribute to the long-term polyuria and impairment of urinary concentrating capacity associated with obstructive nephropathy. PMID- 11399658 TI - Alpha1-adrenoceptor subtypes on rat afferent arterioles assessed by radioligand binding and RT-PCR. AB - We utilized [3H]prazosin saturation and competition radioligand binding studies to characterize the expression of alpha1-adrenoceptors in preglomerular vessels. mRNA for adrenoceptor subtypes was assayed using RT-PCR. The vessels were isolated using an iron oxide-sieving method. [3H]prazosin bound to a single class of binding sites (Kd 0.087 +/- 0.012 nM, Bmax 326 +/- 56 fmol/mg protein). Phentolamine displaced [3H]prazosin (0.2 nM) with a pK(i) of 8.37 +/- 0.09. Competition with the selective alpha1A-adrenoceptor antagonist 5-methylurapidil fit a two-site model (pK(i) 9.38 +/- 0.21 and 7.04 +/- 0.15); 59 +/- 3% of the sites were high-affinity, and 41 +/- 3% were low-affinity binding sites. Competition with the alpha1D-adrenoceptor antagonist 8-(2-[4-(2-methoxyphenyl)-1 piperazinyl]ethyl)-8-azaspiro[4.5]decane-7,9-dione dihydrochloride (BMY-7378) fit a one-site model with low affinity (pK(i) 6.83 +/- 0.03). The relative contents of alpha1A-, alpha1B-, and alpha1D-adrenoceptor mRNAs were 64 +/- 5, 25 +/- 5, and 11 +/- 1%, respectively. Thus there was a very good correlation between mRNA and receptor binding for the subtypes. These data indicate a predominance of the alpha1A-adrenoceptor subtype in rat renal resistance vessels, with smaller densities of alpha1B- and alpha1D-adrenoceptors. PMID- 11399659 TI - Tetrahydrobiopterin reverses the inhibition of nitric oxide by high glucose in cultured murine mesangial cells. AB - Alterations of intrarenal nitric oxide (NO) synthesis play an important role in the pathogenesis and progression of diabetic nephropathy. We tested the hypothesis that hyperglycemia modulates intrarenal NO synthesis, which might mediate the mesangial cell proliferation and matrix production. Murine mesangial cells were grown in media containing varying glucose concentrations, and cytokine induced NO synthesis was assayed by chemiluminescence using an NO analyzer. High media glucose (25 mM) inhibited NO synthesis in a time-dependent fashion. This inhibition was posttranslational as revealed by analysis of inducible nitric oxide synthase (iNOS) gene and protein expression. L-Arginine supplementation partially reversed the inhibition whereas addition of tetrahydrobiopterin (BH4), a cofactor for NOS, restored the inducibility of NO synthesis. The in vitro [3H]citrulline assay for iNOS activity indicated that high glucose decreased BH4 availability whereas examination of the BH4 synthetic pathway suggested decreased BH4 stability rather than synthesis, a defect that was corrected by ascorbic acid. We conclude that hyperglycemia inhibits NO synthesis in mesangial cells by a posttranslational defect that might involve the stability and hence availability of BH4. PMID- 11399660 TI - Real-time profiling of kidney tubular fluid nitric oxide concentrations in vivo. AB - To directly determine intratubular nitric oxide concentrations ([NO]) in vivo, we modified amperometric integrated electrodes (WPI P/N ISO-NOP007), which are highly sensitive to NO and not affected by ascorbic acid, nitrite, L-arginine, or dopamine. Although reactive lengths were as short as 5 microm long, the electrode still responded rapidly. With the use of kidney surface fluid as the "zero point," the electrode tip was inserted into tubular segments along the track of a perforation made by a beveled glass pipette. The surface fluid zero point was usually stable as distal, late proximal, and early proximal tubule [NO] levels were measured sequentially in the same nephron. In eight normal rats, distal, late proximal, and early proximal [NO] concentrations were each approximately 110 nM. In contrast, in nine 5/6 nephrectomized rats 2 wk postsurgery, although [NO] also did not differ among distal, late proximal, and early proximal segments, levels were approximately fourfold higher than those in normal rats and were significantly reduced after N(G)-monomethyl-L-arginine administration. These are the first quantitative in vivo tubular fluid [NO] measurements and show a significant increase in tubular fluid [NO] after renal ablation. PMID- 11399661 TI - Y-27632, an inhibitor of Rho-kinase, antagonizes noradrenergic contractions in the rabbit and human penile corpus cavernosum. AB - We have examined the effect of an inhibitor of Rho-kinase, (+)-(R)-trans-4-(1 aminoethyl)-N-(4-pyridyl) cyclohexanecarboxamide dihydrochloride monohydrate (Y 27632), on the contractions elicited by noradrenergic nerve stimulation and by phenylephrine in the human and rabbit penile corpus cavernosum. In both tissues, after treatment with scopolamine (10 microM) and N(G)-nitro-L-arginine methyl ester (L-NAME; 300 microM), electrical field stimulation (EFS) elicited noradrenergic contractions. These contractions were inhibited by Y-27632 in a concentration-dependent manner. The compound caused concentration-dependent relaxation of phenylephrine-contracted tissues, which were treated with scopolamine (10 microM), guanethidine (10 microM) and L-NAME (300 microM). These results suggest that Rho-kinase is involved in noradrenergic contractile pathway in the cavernosal smooth muscle of the penis. PMID- 11399662 TI - Influence of the 5-HT(2C) receptor antagonist SB242,084 on behaviour produced by the 5-HT(2) agonist Ro60-0175 and the indirect 5-HT agonist dexfenfluramine. AB - Ro60-0175 has been described as a selective agonist at the 5-HT(2C) receptor, yet it has only 10- fold higher affinity at the 5-HT(2C) compared to the 5-HT(2A) subtype, and equivalent affinity for the 5-HT(2B) receptor. The selective 5 HT(2C) receptor antagonist SB242,084 (0.5 mg kg(-1) i.p.), blocked the hypoactivity and penile grooming induced by Ro60-0175 (1 mg kg(-1) s.c.). The combination of SB242,084 (0.5 mg kg(-1) i.p.) and Ro60-0175 (3 - 10 mg kg(-1)) produced a completely different pattern of behaviours including wet-dog shakes, hyperactivity and back muscle contractions. These latter effects were blocked by the selective 5-HT(2A) receptor antagonist MDL100,907 (0.5 mg kg(-1) i.p.), but not the 5-HT(2B) receptor antagonist SB215,505 (3 mg kg(-1) p.o.). The indirect 5 HT releaser/reuptake inhibitor dexfenfluramine (1 - 10 mg kg(-1) i.p.) produced a mild increase in locomotor activity, penile grooming, and occasional back muscle contractions and wet-dog shakes. Pre-treatment with SB242,084 (0.5 mg kg(-1)), blocked the incidence of penile grooming, and markedly potentiated both the dexfenfluramine-induced hyperactivity, the incidence of back muscle contractions, and to a lesser extent wet-dog shakes. Some toxicity was also evident in animals treated with dexfenfluramine (10 mg kg(-1))/SB242,084 (0.5 mg kg(-1)), but not in any other treatment groups. The hyperactivity and toxicity produced by the dexfenfluramine (10 mg kg(-1))/SB242,084 (0.5 mg kg(-1)) combination was replicated in a further study, and hyperthermia was also recorded. Both hyperthermia and toxicity were blocked by MDL100,907 (0.5 mg kg(-1)) but not SB215,505 (3 mg kg(-1)). An attenuation of the hyperlocomotor response was also observed following MDL100,907. These findings suggest that 5-HT(2C) receptor activation can inhibit the expression of behaviours mediated through other 5-HT receptor subtypes. PMID- 11399663 TI - Dexamethasone inhibits TNF-alpha-induced apoptosis and IAP protein downregulation in MCF-7 cells. AB - Exposure of human mammary carcinoma cell line MCF-7 to TNF-alpha leads to apoptotic cell death within 24 h. In search for apoptosis-preventing signals, we identified glucocorticoids as potent death-preventing compounds. Ten nM dexamethasone provided a significant protective effect whereas 100 nM dexamethasone roughly blocked 80 - 90% of TNF-alpha-induced apoptosis. Surprisingly, dexamethasone exerted a protective effect even when supplied several hours after TNF-alpha. This points to a powerful inhibition of even advanced apoptotic processes by dexamethasone. To further pinpoint the anti apoptotic glucocorticoid action, we investigated the expression levels of several members of the inhibitors of apoptosis (IAPs) family of proteins in response to TNF-alpha and dexamethasone. IAP proteins directly block caspase protease activities including caspase-3, caspase-7, and caspase-9. Exposure of MCF-7 cells to TNF caused an extensive downregulation of cIAP1, cIAP2, and XIAP protein levels. The decline of the IAP protein levels temporally paralleled the appearance of apoptotic DNA fragments which started 12 - 14 h following TNF-alpha addition and maximal effects were seen within 24 h. Coincubation of cells with TNF-alpha and dexamethasone potently blocked cIAP1, cIAP2, and XIAP downregulation. TNF-alpha-mediated IAP protein downregulation was not affected by proteasome inhibitors like lactacystin, ALLN or ALLM, whereas it was blocked by the broad-spectrum caspase inhibitor Z-VAD-fmk which also prevented TNF-alpha induced apoptotic cell death. These data suggest that inhibition of IAP downregulation mediated by a caspase proteolytic activity constitutes the anti apoptotic action of glucocorticoids in MCF-7 carcinoma cells. PMID- 11399664 TI - Protection of vascular wall function in insulin-resistant rats from copper oxidative stress. AB - The effects of oxidative stress on vascular function in the insulin-resistant state were assessed in mesenteric resistance arteries of obese, insulin-resistant (cp/cp) and lean, normal (+/?) JCR : LA-cp rats. Nitric oxide-mediated relaxation of noradrenaline-contracted arteries in response to acetylcholine was impaired after 2 h of incubation with Cu(2+) in both genotypes, with or without the continuing presence of Cu(2+). Relaxation was enhanced on initial exposure to Cu(2+), and post-incubation removal of the Cu(2+) resulted in a greater impairment of relaxation. Arteries from cp/cp rats were less impaired in function by Cu(2+) incubation than were those of +/? controls. Sodium nitroprusside mediated relaxation was impaired by exposure to Cu(2+), with an accompanying increase in EC(50). The impairment in acetylcholine-mediated relaxation in the arteries from both cp/cp and +/? rats was completely inhibited by co-incubation with copper-zinc superoxide dismutase and catalase, confirming that the impairment associated with Cu(2+) incubation was due to oxidative stress. The impairment appears to involve both smooth muscle and the endothelium. The cp/cp rats showed greater resistance to the effects of oxidative stress on arterial function, possibly due to an adaptation to oxidative stress on arterial function associated with the insulin-resistant state. PMID- 11399665 TI - Cyclic AMP elevating agents and nitric oxide modulate angiotensin II-induced leukocyte-endothelial cell interactions in vivo. AB - Angiotensin (Ang-II) is a key molecule in the development of cardiac ischaemic disorders and displays proinflammatory activity in vivo. Since intracellular cyclic nucleotides elevating agents have proved to be effective modulators of leukocyte recruitment, we have evaluated their effect on Ang-II-induced leukocyte endothelial cell interactions in vivo using intravital microscopy within the rat mesenteric microcirculation. Pretreatment with iloprost significantly inhibited (1 nM) Ang-II-induced increase in leukocyte rolling flux, adhesion and emigration at 60 min by 96, 92 and 90% respectively, and returned leukocyte rolling velocity to basal levels. Pretreatment with salbutamol or co-superfusion with forskolin exerted similar effects. When theophylline was administered, leukocyte rolling flux, adhesion and emigration elicited by Ang-II were significantly attenuated by 81, 89 and 71% respectively. Rolipram administration caused similar reduction of Ang-II-induced leukocyte responses. Co-superfusion of Ang-II with the NO-donor, spermine-NO, or 8-Br-cyclic GMP, or pretreatment with a transdermal nytroglycerin patch, resulted in a significant reduction of the leukocyte-endothelial cell interactions elicited by Ang-II. Salbutamol preadministration did not modify leukocyte-endothelial cell interactions elicited by either L-NAME or L-NAME+Ang II, indicating that the inhibitory leukocyte effects caused by cyclic AMP elevating agents are mediated through NO release. In conclusion, we have provided evidence that cyclic AMP elevating agents and NO donors, are potent inhibitors of Ang-II-induced leukocyte-endothelial cell interactions. Thus, they could constitute a powerful therapeutical tool in the control of the leukocyte recruitment characteristic of the vascular lesions that occur in cardiovascular disease states where Ang-II plays a critical role. PMID- 11399666 TI - Acute effect of the dual angiotensin-converting enzyme and neutral endopeptidase 24-11 inhibitor mixanpril on insulin sensitivity in obese Zucker rat. AB - The aim of this study was to determine whether acute dual angiotensin-converting enzyme (ACE)/neutral endopeptidase 24-11 (NEP) inhibition could improve whole body insulin-mediated glucose disposal (IMGD) more than ACE inhibition alone and whether this effect was mediated by the kinin-nitric oxide (NO) pathway activation. We therefore compared in anaesthetized obese (fa/fa) Zucker rats (ZOs) the effects of captopril (2 mg kg(-1), i.v.+2 mg kg(-1) h(-1)), retrothiorphan (25 mg kg(-1), i.v. +25 mg kg(-1) h(-1)), a selective NEP inhibitor, and mixanpril (25 mg kg(-1), i.v. +25 mg kg(-1) h(-1)), a dual ACE/NEP inhibitor, on IMGD using hyperinsulinaemic euglycaemic clamp technique. The role of the kinin-NO pathway in the effects of mixanpril was tested using a bradykinin B2 receptor antagonist (Hoe-140, 300 microg kg(-1)) and a NO-synthase inhibitor (N(omega)-nitro-L-arginine methyl ester, L-NAME, 10 mg kg(-1) i.v. +10 mg kg(-1) h(-1)) as pretreatments. Insulin sensitivity index (ISI) was lower in ZO controls than in lean littermates. Increases in ISI were observed in captopril- and retrothiorphan-treated ZOs. In mixanpril-treated ZOs, ISI was further increased, compared to captopril- and retrothiorphan-treated ZOs. In ZOs, Hoe-140 and L-NAME alone did not significantly alter and slightly reduced the ISI respectively. Hoe 140 and L-NAME markedly inhibited the ISI improvement induced by mixanpril. These results show that in obese insulin-resistant Zucker rats, under acute conditions, NEP or ACE inhibition can improve IMGD and that dual ACE/NEP inhibition improves IMGD more effectively than does either single inhibition. This effect is linked to an increased activation of the kinin-NO pathway. PMID- 11399667 TI - Ergolide, sesquiterpene lactone from Inula britannica, inhibits inducible nitric oxide synthase and cyclo-oxygenase-2 expression in RAW 264.7 macrophages through the inactivation of NF-kappaB. AB - We investigated the mechanism of suppression of inducible nitric oxide synthase (iNOS) and cyclo-oxygenase-2 (COX-2) by ergolide, sesquiterpene lactone from Inula britannica. iNOS activity in cell-free extract of LPS/IFN-gamma-stimulated RAW 264.7 macrophages was markedly attenuated by the treatment with ergolide. Its inhibitory effect on iNOS was paralleled by decrease in nitrite accumulation in culture medium of LPS/IFN-gamma-stimulated RAW 264.7 macrophages in a concentration-dependent manner. However, its inhibitory effect does not result from direct inhibition of the catalytic activity of NOS. Ergolide markedly decreased the production of prostaglandin E(2) (PGE(2)) in cell-free extract of LPS/IFN-gamma-stimulated RAW 264.7 macrophages in a concentration-dependent manner, without alteration of the catalytic activity of COX-2 itself. Ergolide decreased the level of iNOS and COX-2 protein, and iNOS mRNA caused by stimulation of LPS/IFN-gamma in a concentration-dependent manner, as measured by Western blot and Northern blot analysis, respectively. Ergolide inhibited nuclear factor-kappaB (NF-kappaB) activation, a transcription factor necessary for iNOS and COX-2 expression in response to LPS/IFN-gamma. This effect was accompanied by the parallel reduction of nuclear translocation of subunit p65 of NF-kappaB as well as IkappaB-alpha degradation. In addition, these effects were completely blocked by treatment of cysteine, indicating that this inhibitory effect of ergolide could be mediated by alkylation of NF-kappaB itself or an upstream molecule of NF-kappaB. Ergolide also directly inhibited the DNA-binding activity of active NF-kappaB in LPS/IFN-gamma-pretreated RAW 264.7 macrophages. These results demonstrate that the suppression of NF-kappaB activation by ergolide might be attributed to the inhibition of nuclear translocation of NF-kappaB resulted from blockade of the degradation of IkappaB and the direct modification of active NF-kappaB, leading to the suppression of the expression of iNOS and COX 2, which play important roles in inflammatory signalling pathway. PMID- 11399668 TI - Effects of pH on the inhibition of fatty acid amidohydrolase by ibuprofen. AB - The pharmacological properties of fatty acid amidohydrolase (FAAH) at different assay pH values were investigated using [(3)H]-anandamide ([(3)H]-AEA) as substrate in rat brain homogenates and in COS-1 [corrected] cells transfected with wild type and mutant FAAH. Rat brain hydrolysis of [(3)H]-AEA showed pH dependency with an optimum around pH 8-9. Between pH 6.3 and 8.2, the difference in activity was due to differences in the V(max), rather than the K(M) values. For inhibition of rat brain [(3)H]-AEA metabolism by a series of known FAAH inhibitors, the potencies of the enantiomers of ibuprofen and phenylmethylsulphonyl fluoride (PMSF) were higher at pH 5.28 than at pH 8.37, whereas the reverse was true for oleyl trifluoromethylketone (OTMK) and arachidonoylserotonin. At both pH values, (-)ibuprofen was a mixed-type inhibitor of FAAH. The K(i)((slope)) and K(i)((intercept)) values for (-)ibuprofen at pH 5.28 were 11 and 143 microM, respectively. At pH 8.37, the corresponding values were 185 and 3950 microM, respectively. The pH dependency for the inhibition by OTMK and (-)ibuprofen was also seen in COS-1 [corrected] cells transiently transfected with either wild type, S152A or C249A FAAH. No differences in potencies between the wild type and mutant enzymes were seen. It is concluded that the pharmacological properties of FAAH are highly pH-dependent. The higher potency of ibuprofen at lower pH values raises the possibility that in certain types of inflamed tissue, the concentration of this compound following oral administration may be sufficient to inhibit FAAH. PMID- 11399669 TI - The C6-2B glioma cell P2Y(AC) receptor is pharmacologically and molecularly identical to the platelet P2Y(12) receptor. AB - P2Y receptor activation in many cell types leads to phospholipase C activation and accumulation of inositol phosphates, while in blood platelets, C6-2B glioma cells, and in B10 microvascular endothelial cells a P2Y receptor subtype, which couples to inhibition of adenylyl cyclase, historically termed P2Y(AC), (P2T(AC) or P(2T) in platelets) has been identified. Recently, this receptor has been cloned and designated P2Y(12) in keeping with current P2 receptor nomenclature. Three selective P(2T) receptor antagonists, with a range of affinities, inhibited ADP-induced aggregation of washed human or rat platelets, in a concentration dependent manner, with a rank order of antagonist potency (pIC(50), human: rat) of AR-C78511 (8.5 : 9.1)>AR-C69581 (6.2 : 6.0)>AR-C70300 (5.4 : 5.1). However, these compounds had no effect on ADP-induced platelet shape change. All three antagonists had no significant effect on the ADP-induced inositol phosphate formation in 1321N1 astrocytoma cells stably expressing the P2Y(1) receptor, when used at concentrations that inhibit platelet aggregation. These antagonists also blocked ADP-induced inhibition of adenylyl cyclase in rat platelets and C6-2B cells with identical rank orders of potency and overlapping concentration - response curves. RT - PCR and nucleotide sequence analyses revealed that the C6 2B cells express the P2Y(12) mRNA. These data demonstrate that the P2Y(AC) receptor in C6-2B cells is pharmacologically identical to the P2T(AC) receptor in rat platelets. PMID- 11399670 TI - The mechanism for the contraction induced by leukotriene C4 in guinea-pig taenia coli. AB - The mechanism underlying the LTC(4)-induced contraction of guinea-pig taenia coli was determined using the simultaneous measurements of [Ca(2+)](i) and force in whole muscle preparations. Additional experiments were performed in receptor coupled permeabilized preparation. For comparison purposes, the contraction which was induced by a typical G-protein mediated agonist, carbachol was also characterized. LTC(4) induced a contraction in the guinea-pig taenia coli in a concentration-dependent manner. The maximal response was obtained at 100 nM and the EC(50) value was 5.4+/-1.9 nM. Both LTC(4) and carbachol induced increases in [Ca(2+)](i) and force. The maximum force induced by 100 nM LTC(4) was significantly smaller than that induced by 10 microM carbachol, although an increase in [Ca(2+)](i) produced by both agonists was similar. In the permeabilized preparations, carbachol, but not LTC(4), induced an additional force development at a fixed Ca(2+) concentration. LTC(4) induced no increase in [Ca(2+)](i) and force in the Ca(2+)-free solution, while carbachol induced transient increases in both [Ca(2+)](i) and force in a Ca(2+)-free solution. Both diltiazem and SK&F 96365 significantly inhibited the LTC(4)- and carbachol induced increases in [Ca(2+)](i) and force in normal PSS. The inhibitory pattern of [Ca(2+)](i) by these drugs was also similar. We thus conclude that LTC(4) induces the contraction of the guinea-pig taenia coli mainly through Ca(2+) influx via both the diltiazem-sensitive and SK&F 96365-sensitive Ca(2+) channels, without affecting either the Ca(2+)-sensitivity or the intracellular Ca(2+) release. These results indicated that the mechanism underlying the LTC(4)-induced contraction differs greatly from that for conventional G-protein mediated agonists, such as carbachol. PMID- 11399671 TI - Effects of GABA(A) receptor partial agonists in primary cultures of cerebellar granule neurons and cerebral cortical neurons reflect different receptor subunit compositions. AB - Based on an unexpected high maximum response to piperidine-4-sulphonic acid (P4S) at human alpha1alpha6beta2gamma2 GABA(A) receptors expressed in Xenopus oocytes attempts to correlate this finding with the pharmacological profile of P4S and other GABA(A) receptor ligands in neuronal cultures from rat cerebellar granule cells and rat cerebral cortex were carried out. GABA and isoguvacine acted as full and piperidine-4-sulphonic acid (P4S) as partial agonists, respectively, at alpha1beta2gamma2, alpha6beta2gamma2 and alpha1alpha6beta2gamma2 GABA receptors expressed in Xenopus oocytes with differences in potency. Whole-cell patch-clamp recordings were used to investigate the pharmacological profile of the partial GABA(A) receptor agonists 4,5,6,7-tetrahydroisoxazolo-(5,4-c)pyridin-3-ol (THIP), P4S, 5-(4-piperidyl)isoxazol-3-ol (4-PIOL), and 3-(4-piperidyl)isoxazol-5-ol (iso 4-PIOL), and the competitive GABA(A) receptor antagonists Bicuculline Methbromide (BMB) and 2-(3-carboxypropyl)-3-amino-6-methoxyphenyl-pyridazinium bromide (SR95531) on cerebral cortical and cerebellar granule neurons. In agreement with findings in oocytes, GABA, isoguvacine and P4S showed similar pharmacological profiles in cultured cortical and cerebellar neurones, which are known to express mainly alpha1, alpha2, alpha3, and alpha5 containing receptors and alpha1, alpha6 and alpha1alpha6 containing receptors, respectively. 4-PIOL and iso-4-PIOL, which at GABA(A) receptors expressed in oocytes were weak antagonists, showed cell type dependent potency as inhibitors of GABA mediated responses. Thus, 4-PIOL was slightly more potent at cortical neurones than at granule neurones and iso-4-PIOL was more potent in inhibiting isoguvacine-evoked currents at cortical than at granule neurons. Furthermore the maximum response to 4-PIOL corresponded to that of a partial agonist, whereas that of iso-4-PIOL gave a maximum response close to zero. It is concluded that the pharmacological profile of partial agonists is highly dependent on the receptor composition, and that small structural changes of a ligand can alter the selectivity towards different subunit compositions. Moreover, this study shows that pharmacological actions determined in oocytes are generally in agreement with data obtained from cultured neurons. PMID- 11399672 TI - Dominant role of L- and P-selectin in mediating CXC chemokine-induced neutrophil migration in vivo. AB - The role of selectins in neutrophil emigration in response to the CXC chemokines KC and MIP-2 was investigated in wild type and P-selectin deficient mice. Intrapleural injection of KC or MIP-2 induced a rapid and specific neutrophil accumulation. Emigration 2 h after KC or MIP-2 was reduced 83 - 88% by anti-L selectin mAb and 53 - 63% by anti-P-selectin mAb. Co-administration of anti-L- and P-selectin mAbs abolished neutrophil migration induced by either chemokine. An anti-E-selectin mAb tested alone did not affect KC-induced neutrophil migration after 2 or 4 h. Moreover, anti-E-selectin did not have an additive inhibitory effect on KC-induced neutrophil migration compared with P-selectin blockade alone. This was found when neutrophil migration was measured at 2 and 4 h after KC. Despite a blood neutrophilia, neutrophil migration at 2 and 4 h after KC was markedly smaller (by approximately 90%) in P-selectin deficient mice compared with wild type animals. Responses at both time points were not decreased further in animals given E-selectin mAb but were reduced to the PBS control level in the presence of anti-L-selectin. In vitro study of cultured murine endothelial cells demonstrated that KC can directly increase cell surface P-selectin expression. These data suggest that CXC chemokine-induced neutrophil accumulation is dependent on both neutrophil L-selectin and a rapid upregulation of endothelial P-selectin but there is no evidence for E-selectin induction. PMID- 11399673 TI - Pharmacological discrimination between effects of carbamazepine on hippocampal basal, Ca(2+)- and K(+)-evoked serotonin release. AB - To elucidate mechanisms of hippocampal serotonin release and possible mechanisms of clinical action of carbamazepine (CBZ), we determined interaction between antagonists of N-type (omega-conotoxin GVIA:GVIA), P-type (omega-agatoxin IVA:IVA) Ca(2+) channels, Na(+) channel (tetrodotoxin: TTX) and CBZ on hippocampal basal, Ca(2+)- and K(+)-evoked serotonin releases, using microdialysis in freely moving rats. Basal release was reduced by TTX, GVIA and IVA (GVIA>IVA). Ca(2+)-evoked release was reduced by GVIA but unaffected by TTX and IVA. K(+)-evoked release was reduced by TTX, GVIA and IVA (GVIA or = 20% with respect to postdiluent value) was completely abolished by a single dose of salmeterol (T(1); protection index [PI] > or = 50% in all subjects), but it was still present after 1 week of regular treatment with salmeterol (T(2); PI < 50% in all subjects). The maximum FEV(1) percentage fall during sBPT with allergen was significantly lower after withdrawal of regular inhaled salmeterol (T(3)) than after regular treatment with salmeterol (T(2)) (mean, 23% vs 29.5%; range, 4 to 41% vs 18 to 49%, respectively; p < 0.05); a similar result was obtained considering the PI of salmeterol on sBPT with allergen (mean, 44% vs 20%; range, 2 to 86% vs - 11 to 49%, respectively; p < 0.05). However, the maximum FEV(1) percentage fall and PI were significantly different in T(3) than after T(1), and only 4 of 10 patients showed in T(3) a PI > or = 50%. CONCLUSIONS: The bronchoprotective effect of salmeterol on allergen-induced EAR, completely lost after 1 week of regular treatment with salmeterol, may be partially restored by the withdrawal of salmeterol therapy for 3 days (72 h). However, this withdrawal time period is not sufficient to recover the baseline bronchoprotective efficacy of the first dose of salmeterol. PMID- 11399690 TI - The prevalence of exercise-induced bronchospasm among US Army recruits and its effects on physical performance. AB - STUDY OBJECTIVES: To measure the prevalence of exercise-induced bronchospasm (EIB) and to determine its effect on the physical performance response to training in otherwise healthy young adults. DESIGN: Observational, retrospective study. SETTING: Fort Jackson, SC, May to July 1998. PARTICIPANTS: One hundred thirty-seven ethnically diverse US Army recruits undergoing an 8-week Army basic training course. MEASUREMENTS AND RESULTS: Subjects underwent exercise challenge testing at the end of basic training to evaluate for EIB (defined as a decrease in FEV(1) of > or = 15%, 1 or 10 min after running to peak oxygen uptake on a treadmill). Those subjects who were unable to run to peak oxygen uptake, or who were unable to perform two baseline FEV(1) maneuvers the results of which were within 5% of each other, were excluded from analysis. We measured peak oxygen uptake on a treadmill and the scores achieved on the components of the US Army physical fitness test (APFT). Of 137 subjects, 121 (58 men and 63 women) met our inclusion criteria. Eight subjects (7%) had EIB. Subjects who experienced EIB and unaffected control subjects both showed statistically significant gains in performance on the APFT events during basic training. At the end of basic training, peak oxygen uptake levels and APFT event scores were not significantly different between subjects with EIB and unaffected control subjects. CONCLUSIONS: Seven percent of the US Army recruits who were tested had EIB, but this did not hinder their physical performance gains during basic training. EIB per se should not be an absolute reason to exclude individuals from employment in jobs with heavy physical demands. PMID- 11399691 TI - Changes in FVC during methacholine-induced bronchoconstriction in elderly patients with asthma: bronchial hyperresponsiveness and aging. AB - STUDY OBJECTIVE: We evaluated whether aging may produce changes in bronchial hyperresponsiveness, risk of enhanced bronchoconstriction, and changes of bronchoconstriction perception. SETTING: Each subject underwent a methacholine bronchial challenge. Methacholine challenge was stopped when one of the following conditions occurred: (1) plateau of bronchoconstriction; (2) decrease of FEV(1) > 40%; (3) FEV(1) drop below 1 L; or (4) excessive respiratory discomfort. Methacholine dose-response curves were plotted both for FVC and FEV(1). The provocative dose of methacholine causing a 20% decrease in FEV(1) with respect to baseline (PD(20)) and the fall in FVC (DeltaFVC) at PD(20) were computed. The Borg scale was used for scoring the perception of respiratory discomfort. PATIENTS: We compared 17 young asthmatic patients (aged 22 to 45 years) with 17 older asthmatic patients (aged 63 to 78 years) selected on the basis of similar baseline pulmonary function and disease duration. RESULTS: No significant between group difference was found in PD(20) and in plateau development. Conversely, DeltaFVC was significantly higher in the older group (mean +/- SD, 15.5 +/- 3.9% vs 11.6 +/- 5.5% in younger patients). In addition, DeltaFVC showed a positive linear relationship with age (p = 0.0026). Elderly subjects were less aware of bronchoconstriction during the methacholine challenge (p = 0.04). CONCLUSIONS: In elderly patients with asthma having comparable pulmonary function and disease duration, bronchial responsiveness is not different from that observed in younger asthmatic patients. Nevertheless, in such patients, an age-related tendency to an enhanced bronchoconstriction and a reduced perception of the degree of bronchoconstriction exist. PMID- 11399692 TI - Gender bias in the diagnosis of COPD. AB - BACKGROUND: COPD is thought to be more prevalent among men than women, a finding usually attributed to higher smoking rates and more frequent occupational exposures of significance for men. However, smoking prevalence has increased among women and there is evidence that women may be more susceptible to the adverse pulmonary function effects of smoking than men. There may also be underdiagnosis and misdiagnosis of COPD in both sexes because objective measures of lung function are underused. OBJECTIVES: We undertook the present study to determine if there is gender bias in the diagnosis of COPD, such that women are less likely than men to receive a diagnosis of COPD. We also attempted to determine if underuse of lung function measurements was a factor in any bias detected. METHODS: We surveyed a random sample of 192 primary-care physicians (96 American and 96 Canadian; 154 men and 38 women) using a hypothetical case presentation and a structured interview. The case of cough and dyspnea in a smoker was presented in six versions differing only in the age and sex of the patient. After presentation of the history and physical findings, physicians were asked to state the most probable diagnosis and to choose the diagnostic studies needed. Physicians were then presented with spirometric findings of moderate or severe obstruction without significant bronchodilator response, and the questions repeated. Finally, the negative outcome of an oral steroid trial was described. RESULTS: Initially, COPD was given as the most probable diagnosis significantly more often for men than women (58% vs 42%; p < 0.05). The likelihood of a COPD diagnosis increased significantly and initial differences between sexes decreased as objective information was provided. After spirometry, COPD diagnosis rates for men and women were 74% vs 66% (p = not significant); after the steroid trial 85% vs 79% (p = not significant). Only 22% of physicians would have requested spirometry after the initial presentation. CONCLUSIONS: In North America, primary care physicians underdiagnosed COPD, particularly in women. Spirometry reduces the risk of underdiagnosis and gender bias but is underused. PMID- 11399693 TI - Is it really useful to repeat outpatient pulmonary rehabilitation programs in patients with chronic airway obstruction? A 2-year controlled study. AB - STUDY OBJECTIVES: To answer the following questions: in patients with chronic airway obstruction (CAO), (1) can pulmonary rehabilitation lead to similar short term gains at successive, yearly interventions, and (2) is there any real clinical or physiologic long-term benefit by yearly repetition of pulmonary rehabilitation programs (PRPs)? DESIGN: Randomized, controlled clinical study. SETTING: Pulmonary rehabilitation center. PATIENTS: Sixty-one CAO patients studied 1 year after completing an initial 8-week outpatient PRP (PRP1). INTERVENTION: Patients were randomly classified into two groups. A second PRP (PRP2) was completed by the first group (group 1) but not by the second group (group 2). One year later, a third PRP (PRP3) was performed by both groups. MEASUREMENTS: Lung function, cycloergometry, walking test, dyspnea, and health related quality of life (HRQL) were assessed before and after PRP2, and before and after PRP3. The numbers of hospitalizations and exacerbations over the year were also recorded. RESULTS: Complete data sets were obtained from 36 patients (17 patients in group 1 and 19 patients in group 2). The two groups did not differ in any parameter either before PRP1, after PRP1, or at randomization. There was no significant change over time for airway obstruction in either group. After PRP2, exercise tolerance, dyspnea, and HRQL improved in group 1. Nevertheless, 1 year later, patients of group 1 did not differ from patients of group 2 in any outcome parameter, such that in comparison to before PRP1, only HRQL was still better in both groups 24 months after PRP1. Yearly hospitalizations and exacerbations per patient significantly decreased in both groups in the 2 years following PRP1, when compared to the 2 years prior. Nevertheless, at the 24-month follow-up visit, a further reduction in yearly exacerbations was observed only in group 1 but not in group 2 in comparison to what was observed at the 12-month follow-up visit. The PRP3 resulted in improvement in exercise tolerance in both groups. CONCLUSION: In patients with CAO, an outpatient PRP can achieve benefits in HRQL and a decreased number of hospitalizations, which persist for a period of 2 years. Successive, yearly interventions lead to similar short-term gains but do not result in additive long term physiologic benefits. Further reduction in yearly exacerbations seems to be the main benefit of an additional PRP. PMID- 11399694 TI - The effectiveness of outpatient pulmonary rehabilitation in chronic lung disease: a randomized controlled trial. AB - BACKGROUND: Patients with chronic pulmonary disease have been shown to benefit from pulmonary rehabilitation programs. Published work has often been from specialized teaching centers and has involved inpatient stay. We assessed an entirely outpatient-based program of pulmonary rehabilitation in patients with chronic lung disease, using the St. George's Respiratory Questionnaire (SGRQ) (which measures health-related quality of life) as the primary outcome measure. METHODS: We undertook a randomized, prospective, parallel-group controlled study of an outpatient rehabilitation program in 65 patients with COPD (44 men and 21 women; mean age, 69.5 years [SD, 9.2 years]; FEV(1), 41% predicted [SD, 18.5%]). The active group (n = 36) took part in a 6-week program of education (2 h weekly) and exercise (1 h weekly). The control group (n = 29) were reviewed routinely as medical outpatients. The SGRQ was administered under supervision by a blinded observer at study entry, 12 weeks, and 24 weeks. RESULTS: The SGRQ in the active group was 59.9 (SE, 2.0) at study entry (n = 36), 47.4 (SE, 2.3) at 12 weeks (n = 32), and 50.6 (SE, 2.5) at 24 weeks (n = 24). The SGRQ in the control group was 59.3 (SE, 2.5) at study entry and did not change significantly over 24 weeks. There was a difference of 10.4 points (confidence interval [CI], 3.6 to 17.3) between the two groups at 12 weeks (p < 0.001) and of 8.1 points (CI, 1.4 to 14.9) at 24 weeks (p = 0.02) in favor of the active group. CONCLUSIONS: A 6-week outpatient-based program significantly improved quality of life in patients with moderate-to-severe COPD. Benefit was still evident after 24 weeks. PMID- 11399695 TI - Geographic variation in long-term oxygen therapy in Denmark : factors related to adherence to guidelines for long-term oxygen therapy. AB - STUDY OBJECTIVES: To evaluate regional differences in adherence to guidelines for long-term oxygen therapy (LTOT) in Denmark and to determine factors related to compliance with these guidelines. DESIGN: Cross-sectional study and analysis of a nationwide database (Danish Oxygen Register). SETTING: Denmark. PATIENTS: In November 1994, 1,354 COPD patients were receiving LTOT in Denmark. MEASUREMENTS AND RESULTS: Among 16 counties, the prevalence of LTOT for COPD varied from 14 to 53 per 100,000. The prevalence was highest in counties where general practitioners (GPs) were prescribing LTOT. Adherence to national guidelines for LTOT was found in 34.4% of the patients for the whole of Denmark and varied regionally from 14 to 63%. Mean compliance with guidelines was 5.3 (range, 2.9 to 9.1) times as likely if the oxygen was prescribed by a pulmonary department compared to LTOT initiated by a GP. CONCLUSIONS: Marked geographic variations in compliance with LTOT guidelines are present even in a small country as Denmark. In general, the adherence to the guidelines is poor, especially when non-chest physicians prescribe LTOT. We therefore recommend that local and national thoracic societies together with health organizations responsible for treatment should play a more forceful role in implementing the guidelines. This could be done by enhanced educational efforts, by monitoring of adherence, or even by centralizing the prescription right to departments with pulmonary physicians. PMID- 11399696 TI - Influenza pneumonia: a descriptive study. AB - OBJECTIVE: To describe the clinical features and complications of patients hospitalized with influenza during the 1999-2000 influenza season. METHODS: We reviewed all cases of patients with influenza admitted to a large metropolitan referral hospital during the 1999-2000 season. RESULTS: Thirty-five adult patients (15 men and 20 women) tested positive for influenza A by direct enzyme immunoassay. A fourfold to sevenfold increase in the number of influenza cases was observed over previous years. Most patients had serious comorbid illnesses (88%), such as diabetes and chronic respiratory and heart disease. Seventeen patients developed pneumonia; these patients tended to be older (mean +/- SD, 63 +/- 13 years vs 51 +/- 19 years, respectively; p = 0.04) and had a higher incidence of chronic lung disease (41% vs 6%, respectively; p = 0.02) than the patients without pneumonia. Shortness of breath was the only symptom that distinguished patients with pneumonia from those with an upper respiratory tract illness alone. Antiviral treatment was begun 4 +/- 3 days from initiation of symptoms in patients with pneumonia and consisted of oseltamivir (35.2%), rimantadine (5.8%), or a combination of both (17.6%). Respiratory and/or blood culture results were positive in five patients (29%), Staphylococcus aureus was isolated in five patients, and Streptococcus pneumoniae was isolated in one patient. Ten of the patients with pneumonia (58.8%) were admitted to the ICU, and 5 patients (29%) died. The length of ICU stay and mechanical ventilation were 28 +/- 26 days and 21.5 +/- 20.5 days, respectively. Death in most pneumonia patients was attributed to respiratory failure. CONCLUSION: The recognized number of hospital admissions for influenza increased fourfold to sevenfold over previous years, most likely due to the implementation of rapid diagnostic tests for influenza. Patients with signs and symptoms of influenza and shortness of breath should undergo chest radiography. Hospitalization of patients with influenza pneumonia occurred in both previously healthy and immunocompromised patients and had a high mortality. S aureus was the most common bacterial isolate in patients with influenza pneumonia. Empiric antibiotics with staphylococcal activity should be used pending culture results in patients with influenza pneumonia. The effectiveness of oseltamivir and rimantadine in treating patients with influenza pneumonia remains to be determined. PMID- 11399697 TI - BAL induces an increase in peripheral blood neutrophils and cytokine levels in healthy volunteers and patients with pneumonia. AB - STUDY OBJECTIVES: To examine the peripheral effects of BAL on the neutrophil counts and cytokine levels in the circulation. DESIGN AND METHODS: WBC counts and plasma cytokines were measured before and 4 h after fiberoptic bronchoscopy (FOB) without further interventions (n = 6), or combined with BAL in normal volunteer subjects (n = 6), and in patients with bacterial pneumonia (n = 4). The bronchus of the right middle lobe was wedged, and three 50-mL aliquots of sterile saline solution was instilled. There was no endotoxin contamination in the saline solution or the fluid obtained through the working channel of bronchoscope. RESULTS: In volunteers, peripheral WBC counts and the number of nonsegmented and segmented neutrophils increased after the BAL procedure (p < 0.05) associated with the increase in plasma concentration (mean +/- SEM) of interleukin (IL)-6 (0.99 +/- 0.32 pg/mL before BAL and 20.38 +/- 13.42 pg/mL after BAL; p < 0.05) and granulocyte colony-stimulating factor (G-CSF; 14.1 +/- 1.7 pg/mL before BAL and 38.5 +/- 9.7 pg/mL after BAL; p < 0.05). The increase in WBC counts and neutrophil counts was positively correlated to the increase in IL-6 (p < 0.05) and the increase in G-CSF (p < 0.05). In patients with pneumonia, IL-6 and G-CSF levels were higher after BAL than in normal volunteer subjects (p < 0.05). There was no increase in plasma concentration of IL-1beta, tumor necrosis factor-alpha, or IL-8 after BAL in normal volunteer subjects or in patients with pneumonia. FOB without BAL did not increase the WBC count, neutrophil count, or plasma cytokine levels. CONCLUSION: The BAL procedure increases the number of WBCs, and segmented and nonsegmented neutrophils in the peripheral circulation as well as circulating IL-6 and G-CSF levels. PMID- 11399698 TI - Treatment of isoniazid-resistant tuberculosis in southeastern Texas. AB - BACKGROUND: Isoniazid-resistant tuberculosis (INHr-TB) can be treated successfully with several treatment regimens. However, the optimal regimen and duration are unclear. STUDY OBJECTIVE: To analyze the efficacy of treatment regimens used for INHr-TB in the southeastern Texas region. DESIGN: Retrospective cohort study. SETTING: Health-care facilities reporting tuberculosis (TB) patients in the Houston and Tyler areas. SUBJECTS: All patients reported to have INHr-TB from 1991 to 1998. Exclusion criteria included poor compliance, additional first-line drug-resistance (except aminoglycosides), and death before completion of 1 month of treatment. MEASUREMENTS AND RESULTS: Main treatment outcomes were treatment failure, relapse, and TB-related death. Fifty-three of 83 patients were included in the study; aminoglycoside resistance coexisted in 37.5% of isolates. Seven types of treatment regimens were identified. Eighteen patients (34%) received rifampin, pyrazinamide, and ethambutol thrice weekly for 9 months. Four patients (7.5%) had a total effective treatment duration of < 9 months. Thirty patients (56.6%) and 16 patients (30.2%) received thrice-daily and daily treatment regimens, respectively. Forty-nine patients achieved sputum conversion. Treatment failure and death occurred in one patient (1.9%). Three patients (5.7%) experienced relapses. There was a significant difference in total effective treatment time between patients with and without relapses (8.3 +/- 1.1 months vs 11.1 +/- 2.1 months; p < 0.02). Twice-weekly treatment regimens were associated with relapse (p = 0.05). CONCLUSIONS: Several treatment regimens were prescribed for INHr-TB in southeastern Texas. INHr-TB treatment durations were > 7 months, and treatment regimen efficacy was adequate. Twice-weekly treatment was associated with relapse, whereas thrice-weekly and daily treatments performed similarly. A prospective study with different treatment durations is needed to determine the optimal treatment regimen for patients with INHr-TB. PMID- 11399699 TI - Evaluation of polymerase chain reaction for detection of Mycobacterium tuberculosis in pleural fluid. AB - OBJECTIVES: Tuberculosis, a reemergent killer, is threatening to assume serious proportions all over the world, particularly in view of the AIDS pandemic. The detection of mycobacterial DNA by polymerase chain reaction (PCR) in clinical samples is a promising approach for the rapid diagnosis of tuberculous infections. The aims of this study were to evaluate PCR for detection of Mycobacterium tuberculosis in pleural fluids and to correlate the results with adenosine deaminase activity (ADA) estimation and acid-fast bacilli (AFB) screening. METHODS: The sensitivity and specificity of PCR in detection of mycobacterial DNA in 20 samples of tuberculous pleural effusion were evaluated using 40 samples of nontubercular pleural effusion as controls. The results were correlated with the ADA in all 60 pleural fluids. In addition, AFB detection by Ziehl-Neelsen staining on cytospin smears of all pleural fluids was also compared. RESULTS: Of the 20 samples of tuberculous pleural effusion, mycobacterium could be detected by AFB staining in 4 samples. Fourteen samples were PCR positive. None of the samples from the control group were AFB or PCR positive. The sensitivity of PCR, therefore, was 70.0% with specificity of 100% (positive predictive value, 100%; negative predictive value, 86.95%). The sensitivity of AFB screening was at best 20%. The mean of ADA values in tubercular pleural effusions was 63.21 U/L (SD, 33.01), and the mean in the control samples was 51.1 U/L (SD, 29.71). Taking a cut-off value of 50 U/L, both the sensitivity and specificity of ADA estimation in diagnosing tuberculosis were only 55%. CONCLUSION: PCR represents a rapid and sensitive method for the detection of mycobacterial DNA in tuberculous pleural effusions. AFB screening has low sensitivity, and ADA estimation has both low sensitivity and specificity. Therefore, when the clinical suspicion is high and smear result is negative, but the signs and symptoms of M tuberculosis are apparent, PCR is the method of choice for identifying the infection. PMID- 11399700 TI - Safety and efficacy of two courses of OM-85 BV in the prevention of respiratory tract infections in children during 12 months. AB - BACKGROUND: Acute respiratory tract infections (ARTIs) are among the main causes of morbidity and mortality in children. The bacterial extract OM-85 BV (bronchovaxom) has shown protective effect for ARTIs on children. We report a double-blind, placebo-controlled, parallel, prospective clinical trial to assess the safety and efficacy of two courses of OM-85 BV in the prevention of ARTIs in susceptible children during 12 months. METHODS: Fifty-four susceptible children from 1 to 12 years of age living in the metropolitan area of Chihuahua City were selected. They were randomized to receive either OM-85 BV or placebo (one capsule a day for 10 days a month for 3 consecutive months) at the beginning of the trial and 6 months later with the same schedule. Patients were followed up for 12 months, including the administration period. The trial began in July 1997 and ended in April 1999. RESULTS: The number (mean +/- SD) of ARTIs was 5.04 +/- 1.99 (median, 5.0) in the OM-85 BV group vs 8.0 +/- 2.55 (median, 8.0) in the placebo group, with a mean difference of - 2.96 (95% confidence interval [CI], - 4.22 to 1.7). The number of antibiotic courses was 2.46 +/- 2.08 (median, 1.5) in the treatment group vs 4.46 +/- 2.08 (median, 4.0) in the control group, a difference of - 2.0 (95% CI, - 3.14 to - 0.86). The total duration of ARTIs was 35.23 +/- 17.64 days (median, 30.5 days) in the OM-85 BV group vs 60.75 +/- 25.44 days (median, 55.0 days) in the placebo group, ie, a difference of - 25.52 days (95% CI, - 37.56 to - 13.47 days), p < 0.001 by Student's t test and Mann-Whitney U test for all the items. Four patients in the OM-85 BV group had five adverse events. Only one episode of skin rash was related to the medication intake. Six patients in the control group had six adverse events. CONCLUSIONS: OM-85 BV had a preventive effect on ARTI in the susceptible children for 12 months with an important reduction on the antibiotic requirements and the number of days of suffering ARTIs. PMID- 11399701 TI - Urinary leukotriene E(4) excretion during the first month of life and subsequent bronchopulmonary dysplasia in premature infants. AB - BACKGROUND: Inflammation plays an important role in the pathogenesis of bronchopulmonary dysplasia (BPD), but the exact nature of this inflammatory process is incompletely understood. Older infants with established BPD have higher levels of urinary leukotriene E(4) (LTE(4)) compared to healthy infants of the same age. This suggests that cysteinyl leukotrienes may play a role in the abnormalities seen in BPD. OBJECTIVES: To measure urinary LTE(4) levels during the first month of life in premature infants, and to determine whether there are significant differences in premature infants who develop BPD, as compared to those who do not develop BPD. DESIGN: Prospective, blinded, controlled study. SETTING: Neonatal ICUs of a tertiary-care university hospital. METHODS: Thirty seven premature infants (< 33 weeks of gestational age) were enrolled prospectively at birth. Urinary LTE(4) levels were measured blinded, using a standard radioimmunoassay technique at 2 days, 7 days, and 28 days of life. At 1 month of age, infants were classified as with or without BPD, based on need for supplemental oxygen, and characteristic chest radiographs. Clinical features and urinary LTE(4) were compared between the two groups. RESULTS: Mean +/- SD gestational age was 29 +/- 2.6 weeks. None of the infants had a family history of asthma. Thirteen of 37 infants were classified as having BPD at 28 days after birth. Mean gestational age in infants who developed BPD was 27 +/- 2.4 weeks, compared to 30 +/- 2 weeks in infants who did not develop BPD (p < 0.05). In infants with BPD, mean urinary LTE(4) levels of urinary creatinine were 1,762 +/- 2,003 pg/mg, 1,236 +/- 992 pg/mg, and 5,541 +/- 5,146 pg/mg at days 2, 7, and 28, respectively, compared to 1,304 +/- 1,195 pg/mg, 1,158 +/- 1,133 pg/mg, and 2,800 +/- 2,080 pg/mg in infants without BPD. LTE(4) levels at 2 days, 7 days, and 28 days did not correlate with the subsequent development of BPD. LTE(4) levels at day 28 were significantly higher than LTE(4) levels at day 2 and day 7 in both groups, even after correcting for gestational age or birth weight (p < 0.05). There was significant inverse correlation between LTE(4) levels at day 2 with gestational age and birth weight (p < 0.05). All 13 infants with BPD received steroid pulses, compared to 3 of 26 infants without BPD. Gestational age and use of postnatal steroid pulses, diuretics, and theophylline (for apnea of prematurity) were significantly associated with each other and with the subsequent development of BPD. CONCLUSION: Urinary LTE(4) levels measured on the second day of life in very-low-birth-weight infants inversely correlate with gestational age and birth weight. Urinary LTE(4) levels may reflect lung injury and/or inflammation in premature infants, not necessarily related to BPD as it is presently defined. PMID- 11399702 TI - Angiotensin-converting enzyme inhibitor therapy improves respiratory muscle strength in patients with heart failure. AB - STUDY OBJECTIVES: Respiratory muscle strength has been shown to be reduced in patients with chronic heart failure. The purpose of this prospective study was to determine whether long-term therapy with the angiotensin-converting enzyme (ACE) inhibitor perindopril improves respiratory muscle strength in patients with chronic heart failure. PATIENTS AND METHODS: Eighteen patients with stable chronic heart failure were administered perindopril, 4 mg/d, in addition to their standard therapy for a period of 6 months. Fourteen patients completed the study. Maximum inspiratory pressure (PImax) and maximum expiratory pressure (PEmax) expressed in percentage of predicted values, left ventricular ejection fraction (LVEF) determined by means of two-dimensional echocardiography, and pulmonary volumes were obtained before and after therapy. MEASUREMENTS AND RESULTS: As compared to baseline, there was a significant increase in both PImax and PEmax after therapy (57 +/- 27% predicted vs 78 +/- 36% predicted and 62 +/- 20% predicted vs 73 +/- 15% predicted, respectively; each p < 0.05). LVEF increased (34 +/- 5% vs 41 +/- 10%; p < 0.05); functional class improved by > or = 1 New York Heart Association (NYHA) class in five patients. There were no changes in pulmonary volumes. No correlation was found between changes in PImax and PEmax and changes in either LVEF or NYHA functional class. CONCLUSIONS: In patients with chronic heart failure, long-term therapy with the ACE inhibitor perindopril improved respiratory muscle strength, as indicated by significant increases in PImax and PEmax. PMID- 11399703 TI - Impaired left ventricular filling due to right ventricular pressure overload in primary pulmonary hypertension: noninvasive monitoring using MRI. AB - OBJECTIVE: To analyze the effect of primary pulmonary hypertension (PPH) on cardiac function using MRI. METHODS: In 12 patients (9 women; age range, 30 to 56 years), the diagnosis of PPH had been established by catheterization (mean +/- SD pulmonary artery pressure [PAP] was 56 +/- 8 mm Hg). With breath-hold cine MRI, a series of short-axis images was acquired covering the whole left ventricle (LV) and right ventricle (RV). The curvature, defined as 1 divided by the radius of curvature in centimeters, was calculated for the septum and the LV free wall in early diastole. Leftward ventricular septal bowing (LVSB) is denoted by a negative curvature. For the LV and the RV, the end-diastolic volume (EDV), stroke volume (SV), and volumetric filling rate were calculated. The control subjects were all healthy (n = 14; 11 women; age range, 20 to 57 years). RESULTS: In the patients, LVSB was quantified in early diastole by the septal curvature of - 0.14 +/- 0.07 cm(-1), and the septal to free-wall curvature ratio of - 0.42 +/- 0.21. LV EDV and LV SV correlated negatively with diastolic PAP (p = 0.004 and p = 0.04, respectively). In patients vs control subjects, RV SV was reduced (52 +/- 12 mL vs 82 +/- 11 mL, p < 0.0001); LV peak filling rate was smaller (2.2 +/- 0.7 EDV/s vs 3.3 +/- 0.5 EDV/s, p < 0.001); LV EDV was smaller (81 +/- 23 mL vs 117 +/- 19 mL, p = 0.001); and LV SV was smaller (49 +/- 18 mL vs 83 +/- 13 mL, p < 0.0001). CONCLUSION: In PPH, RV pressure overload leads to LVSB and reduced RV output. By decreased blood delivery, LV filling is reduced, which results in decreased LV SV by the Frank-Starling mechanism. PMID- 11399704 TI - Dobutamine echocardiography in patients with aortic stenosis and left ventricular dysfunction: predicting outcome as a function of management strategy. AB - STUDY OBJECTIVE: To prospectively address the question whether the assessment of valvular hemodynamics and myocardial function during low-dose dobutamine infusion can guide decision making in patients with aortic stenosis and left ventricular (LV) dysfunction. PATIENTS AND MEASUREMENTS: Twenty-four patients with aortic stenosis and LV dysfunction (mean ejection fraction, 28%; New York Heart Association class, II to IV) were studied by dobutamine echocardiography assessing mean pressure gradient, aortic valve area, and aortic valve resistance. Patients were prospectively divided into severe and nonsevere aortic stenosis groups according to the response of the valve area to the augmentation of systolic flow. The clinical decision was considered to be concordant with the results of dobutamine echocardiography, when patients with severe aortic stenosis and preserved contractile function were referred by a specialist for aortic valve replacement and when patients with nonsevere aortic stenosis were not. Patients were observed for up to 3 years. RESULTS: All eight patients with severe aortic stenosis who were referred for surgery survived and had good cardiovascular outcomes, and six of eight patients who were not initially referred for surgery had poor outcomes, including heart failure and sudden cardiac death. The eight patients with nonsevere aortic stenosis did comparatively well without valve replacement. Cardiac death or pulmonary edema occurred in 4 of 16 patients (25%) when the clinical decision was concordant with the results of the dobutamine echocardiogram and occurred in 6 of 8 patients (75%) when the clinical decision was discordant (p = 0.019 [chi(2) test]). CONCLUSION: Patients with aortic stenosis, LV dysfunction, and relatively low gradients have better outcomes when management decisions are based on the results of dobutamine echocardiograms. Those patients identified as having severe aortic stenosis and preserved contractile reserve by dobutamine echocardiography should undergo surgery, while patients identified as having nonsevere aortic stenosis can be managed conservatively. PMID- 11399705 TI - Differential transesophageal echocardiographic diagnosis between linear artifacts and intraluminal flap of aortic dissection or disruption. AB - BACKGROUND: The relatively low specificity of transesophageal echocardiography (TEE) for the diagnosis of aortic dissection (AD) or traumatic disruption of the aorta (TDA) has been attributed to linear artifacts. We sought to determine the incidence of intra-aortic linear artifacts in a cohort of patients with suspected AD or TDA, to establish the differential TEE diagnostic criteria between these artifacts and true aortic flaps, and to evaluate their impact on TEE diagnostic accuracy. METHODS AND RESULTS: During an 8-year period, patients at high risk of AD (n = 261) or TDA (n = 90) who underwent a TEE study and had confirmed final diagnoses were studied. In an initial retrospective series, linear artifacts were observed within the ascending and descending aorta in 59 of 230 patients (26%) and 17 of 230 patients (7%), respectively. TEE findings associated with linear artifacts in the ascending aorta were as follows: displacement parallel to aortic walls; similar blood flow velocities on both sides; angle with the aortic wall > 85 degrees; and thickness > 2.5 mm. Diagnostic criteria of reverberant images in the descending aorta were as follows: displacement parallel to aortic walls, overimposition of blood flow, and similar blood flow velocities on both sides of the image. In a subsequent prospective series (n = 121), systematic use of these diagnostic criteria resulted in improved TEE specificity for the identification of true intra-aortic flaps. CONCLUSIONS: Misleading intra-aortic linear artifacts are frequently observed in patients undergoing a TEE study for suspected AD or TDA. Routine use of the herein-proposed diagnostic criteria promises to further improve TEE diagnostic accuracy in the setting of severely ill patients with potential need for prompt surgery. PMID- 11399706 TI - Strategies incorporating spiral CT for the diagnosis of acute pulmonary embolism: a cost-effectiveness analysis. AB - OBJECTIVE: To assess the cost-effectiveness of spiral CT for the diagnosis of acute pulmonary embolism. DESIGN: Computer-based cost-effectiveness analysis. PATIENTS: Simulated cohort of 1,000 patients with suspected acute pulmonary embolism (PE), with a prevalence of 28.4%, as in the Prospective Investigation of Pulmonary Embolism Diagnosis study. INTERVENTIONS: Using a decision-analysis model, seven diagnostic strategies were compared, which incorporated combinations of ventilation-perfusion (V/Q) scans, duplex ultrasound of the legs, spiral CT, and conventional pulmonary angiography. MEASUREMENTS AND RESULTS: Expected survival and cost (in Canadian dollars) at 3 months were estimated. Four of the strategies yielded poorer survival at higher cost. The three remaining strategies were as follows: (1) V/Q +/- leg ultrasound +/- spiral CT, with an expected survival of 953.4 per 1,000 patients and a cost of $1,391 per patient; (2) V/Q +/ leg ultrasound +/- pulmonary angiography (the "traditional" algorithm), with an expected survival of 953.7 per 1,000 patients and a cost of $1,416 per patient; and (3) spiral CT +/- leg ultrasound, with an expected survival of 958.2 per 1,000 patients and a cost of $1,751 per patient. The traditional algorithm was then excluded by extended dominance. The cost per additional life saved was $70,833 for spiral CT +/- leg ultrasound relative to V/Q +/- leg ultrasound +/- spiral CT. CONCLUSIONS: Spiral CT can replace pulmonary angiography in patients with nondiagnostic V/Q scan and negative leg ultrasound findings. This approach is likely as effective as-and possibly less expensive than-the current algorithm for diagnosis of acute PE. When spiral CT is the initial diagnostic test, followed by leg ultrasound, expected survival improves but costs are also considerably higher. These findings were robust to variations in the assumed sensitivity and specificity of spiral CT. PMID- 11399707 TI - Primary pulmonary hypertension in Israel: a national survey. AB - OBJECTIVES: To characterize the incidence of patients with primary pulmonary hypertension (PPH) in Israel and their outcomes. METHODS: We have evaluated retrospectively all the patients in Israel in whom PPH was diagnosed between the years 1988 and 1997. We looked at medical history, hemodynamic data, pulmonary function and gas exchange, and demographic variables. Patients were followed up for survival until November 1997. Life table analysis and Kaplan-Meier statistics were used to estimate the overall survival distribution. Regression analysis was used to examine the relations between survival and selected variables. RESULTS: Overall, we found 44 patients with PPH. The estimated incidence of PPH in Israel is 1.4 new cases per year per million population. The mean (+/- SD) age at diagnosis was 43 +/- 13 years. In the Jewish population, PPH was more frequent among immigrants from Europe and the United States. The mean interval from the onset of symptoms to diagnosis was 3 years (median, 2 years). The median survival time was 4 years. The 1-year, 3-year, and 5-year survival rates were 82%, 57%, and 43%, respectively. The major variables influencing the survival rate were the following: interval from symptom onset to diagnosis; and hemodynamic measurements (ie, mean pulmonary artery pressure, mean right atrial pressure, and cardiac index). In comparison to rates discerned from the National Institutes of Health registry data, the survival rate in Israel is somewhat better and prognosis is influenced by similar hemodynamic variables. CONCLUSION: PPH is a rare and fatal disease in Israel. New therapeutic modalities such as prostacyclin therapy and lung transplantation may improve survival among patients with this malignant disease. PMID- 11399708 TI - Slow-wave activity in sleep apnea patients before and after continuous positive airway pressure treatment: contribution to daytime sleepiness. AB - STUDY OBJECTIVES: To estimate the course of slow-wave activity (SWA), its amount during the night, and its correlation with daytime sleepiness in sleep apnea syndrome (SAS) patients. This study also verified whether continuous positive airway pressure (CPAP) treatment also restores a normal pattern of SWA in severe SAS patients. PARTICIPANTS: Ten patients with a diagnosis of severe SAS who showed a good clinical response to CPAP after approximately 9 months of treatment were included in this study. These patients were matched for sex and age with 10 control subjects. DESIGN: All subjects underwent 1 night of polysomnography (PSG), followed by the multiple sleep latency test (MSLT) the next day. For the SAS patients only, the same procedure was repeated after 9 +/- 0.7 months of CPAP treatment. In addition to traditional scoring of sleep stages, apneas, hypopneas, and microarousals, the SWA, defined as the power in the 0.75- to 4.5-Hz frequency band, was evaluated. RESULTS: A positive correlation between SWA of the first cycle and the MSLT (r = 0.56; p = 0.045) was found before treatment. Moreover, SAS patients significantly increased their mean SWA after CPAP treatment in the first (p = 0.024) and second (p = 0.002) sleep cycles and restored a more physiologic decay of SWA across the night. CONCLUSIONS: These results suggest that daytime sleepiness in SAS patients may be the result of a lack of SWA during the first part of the night, and show that CPAP restores a more physiologic pattern of SWA across the night. PMID- 11399709 TI - Treatment effects on carbon dioxide retention in patients with obstructive sleep apnea-hypopnea syndrome. AB - OBJECTIVES: This study was designed to examine respiratory control in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS), with or without CO(2) retention. METHODS: We recruited 10 body mass index-matched, apnea-hypopnea index matched, age-matched, and lung function-matched OSAHS patients, according to their awake PaCO(2). Five patients were hypercapnic (PaCO(2), > or = 45 mm Hg), and five patients were eucapnic. Hypoxic responses (the ratio of the change in minute ventilation [DeltaV(E)] to the change in arterial oxygen saturation [DeltaSaO(2)] and the ratio of the change in mouth occlusion pressure over the first 100 ms of inspiration against an occluded airway [DeltaP(0.1)] to DeltaSaO(2)) and hypercapnic responses (DeltaV(E)/DeltaPCO(2) ratio and DeltaP(0.1)/DeltaPCO(2) ratio) were tested during wakefulness before treatment in all 10 patients, and before and during treatment (at 2, 4, and 6 weeks) with pressure support in the hypercapnic group. RESULTS: Hypercapnic patients had lower mean (+/- SD) DeltaV(E)/DeltaSaO(2) ratio than eucapnic patients (-0.17 +/- 0.04 vs -0.34 +/- 0.04 L /min/%SaO(2), respectively), lower mean DeltaP(0.1)/DeltaSaO(2) ratio (-0.04 +/- 0.02 vs -0.14 +/- 0.03 cm H(2)O/%SaO(2), respectively), and lower DeltaP(0.1)/DeltaPCO(2) ratio (0.23 +/- 0.1 vs 0.49 +/- 0.1 cm H(2)O/mm Hg, respectively) [p < 0.05]. After receiving noninvasive ventilation treatment, the hypercapnic and hypoxic responses of the hypercapnic patients increased. At 4 to 6 weeks, values for both responses had increased to within the normal range and PaCO(2) had fallen to < 45 mm Hg, while weight was unchanged. CONCLUSIONS: Depressed chemoresponsiveness plays a role that is independent of obesity in the development of CO(2) retention in some OSAHS patients, and it may be a response to sleep-disordered breathing. PMID- 11399710 TI - Underestimation of nocturnal hypoxemia due to monitoring conditions in patients with COPD. AB - STUDY OBJECTIVES: COPD patients run a risk of developing nocturnal oxygen desaturation. When evaluating patients with nocturnal hypoxemia, an unfamiliar hospital environment and the monitoring equipment may cause sleep disturbances. It was hypothesized that increased sleep disruption will lead to fewer instances of desaturation during a night of monitoring. DESIGN: The following forms of monitoring were evaluated prospectively on 3 nights for each patient: oximetry at home; polysomnography (PSG) at home; and PSG in the hospital. SETTING: Department of Pulmonology, Rijnstate Hospital Arnhem, The Netherlands. PATIENTS: Fourteen stable COPD patients (7 men; median age, 71.5 years; age range, 59 to 81 years; FEV(1), 32.5% predicted; FEV(1) range, 19 to 70% predicted) participated in the study. All subjects had significant instances of nocturnal arterial oxygen desaturation. Those patients with a sleep-related breathing disorder or cardiac failure were excluded from the study. MEASUREMENTS AND RESULTS: The mean nocturnal arterial oxygen saturation (SaO(2)) level was higher during PSG monitoring at home (89.7%; range, 77 to 93%) than during oximetry monitoring (88.5%; range, 80 to 92%) [p < 0.025]. The fraction of time spent in hypoxemia (ie, SaO(2) < 90%) was lower during PSG monitoring at home (40.8%; range, 5 to 100%) than during oximetry monitoring (59.9%; range, 6 to 100%) [p < 0.01]. Desaturation time (DeltaSaO(2) > 4%) was lower during PSG monitoring at home (22.1%; range, 3 to 63%) during PSG monitoring at home than during oximetry monitoring (50.4%; range, 4 to 91%) [p < 0.01]. A correction for actual sleep during PSG monitoring reduced the differences between PSG monitoring at home and oximetry monitoring, although a difference in the desaturation time remained (PSG monitoring at home, 31.9% [range, 2 to 75%]; oximetry monitoring, 50.4% [range, 4 to 91%]) [p = 0.041]. A comparison of sleep architectures for nights when PSG was being monitored showed a higher arousal index in the hospital than at home (PSG monitoring in the hospital, 5.6 arousals per hour [range, 2 to 16 arousals per hour]; PSG monitoring at home, 2.5 arousals per hour [range, 1 to 6 arousals per hour]) [p < 0.025], but no differences in SaO(2) levels were found between PSG monitoring at home and PSG monitoring in the hospital. CONCLUSION: The artifacts due to sleep-monitoring equipment may cause an underestimation of the degree of nocturnal hypoxemia in COPD patients. The addition of an unfamiliar environment causes more sleep disruption, but this does not affect nocturnal SaO(2) levels further. PMID- 11399711 TI - Hemodynamic effects of simulated obstructive apneas in humans with and without heart failure. AB - STUDY OBJECTIVES: To determine whether generation of negative intrathoracic pressure during apnea would cause more pronounced and sustained reductions in cardiac output in patients with congestive heart failure (CHF) than in healthy subjects. DESIGN: Physiologic intervention study. SETTING: Cardiorespiratory physiology laboratory. PARTICIPANTS: Nine patients with CHF and nine healthy control subjects matched for age and sex. INTERVENTIONS: Patients with CHF and healthy subjects generated - 30 cm H(2)O of intrathoracic pressure during 15-s Mueller maneuvers (MMs) to simulate the acute hemodynamic effects and aftereffects of obstructive apneas. RESULTS: In both groups, MMs caused an immediate rise in left ventricular transmural pressure during systole (LVPtmsys) [p < 0.05], but in CHF patients, this immediate increase was followed by a significant drop in LVPtmsys (p < 0.05), associated with significantly greater reductions in systolic BP and cardiac index than in healthy subjects (- 25 +/- 3 mm Hg vs - 11 +/- 2 mm Hg [p < 0.05] and - 0.53 +/- 0.11 L/min/m(2) vs - 0.15 +/- 0.11 L/min/m(2) [p < 0.05], respectively). Healthy subjects recovered promptly, but in CHF patients, these adverse hemodynamic effects were sustained following release of the MM. CONCLUSIONS: CHF patients experience more pronounced and sustained reductions in BP and cardiac output both during and following the MM than do healthy subjects. These findings suggest the potential for adverse hemodynamic effects and aftereffects of negative intrathoracic pressure generation during obstructive sleep apnea in patients with CHF. PMID- 11399712 TI - Row-a-boat phenomenon: respiratory compensation in advanced Duchenne muscular dystrophy. AB - BACKGROUND: "Row-a-boat" phenomenon (RBP) is a spontaneous upper-body movement in patients with advanced Duchenne muscular dystrophy (DMD), when sitting upright supported by a belt around the body in a wheelchair. However, the role of RBP has not been clarified. OBJECTIVES: To support the hypothesis that RBP is an abnormal pattern of respiration to compensate for the atrophied respiratory muscles in advanced DMD. PATIENTS AND METHODS: Age, degree of ventilator dependency, and blood gas and spirometry values of 12 patients with spontaneous RBP were compared to those of 8 patients without RBP. All patients were men, and all exhibited a comparable level of motor function (unable to ambulate). Spirometry was undertaken with an ambulatory pneumotachograph in six patients with RBP in two conditions: sitting with RBP and sitting without RBP. In the latter condition, because a patient's shoulders, neck, and head were manually restricted, RBP was prevented. RESULTS: We found that the patients with RBP were older (mean, 25.98 years vs 19.84 years), more dependent on mechanical ventilation (13.96 h/d vs 4.31 h/d), and had lower FVC and percentage of FVC (511.3 mL vs 762.5 mL and 13.37% vs 20.11%, respectively) than those without RBP. We also found that the frequency of RBP was identical with tidal breathing, and FVC was increased by 50.8% by simply allowing RBP. CONCLUSION: We conclude that RBP is a respiratory movement to compensate for the atrophied respiratory muscles in advanced DMD. PMID- 11399713 TI - Predictors of outcome for patients with COPD requiring invasive mechanical ventilation. AB - INTRODUCTION: Accurate outcomes data and predictors of outcomes are fundamental to the effective care of patients with COPD and in guiding them and their families through end-of-life decisions. DESIGN: We conducted a retrospective cohort study of 166 patients using prospectively gathered data in patients with COPD who required mechanical ventilation for acute respiratory failure of diverse etiologies. RESULTS: The in-hospital mortality rate for the entire cohort was 28% but fell to 12% for patients with a COPD exacerbation and without a comorbid illness. Univariate analysis showed a higher mortality rate among those patients who required > 72 h of mechanical ventilation (37% vs 16%; p < 0.01), those without previous episodes of mechanical ventilation (33% vs 11%; p < 0.01), and those with a failed extubation attempt (36% vs 7%; p = 0.0001). With multiple logistical regression, higher acute physiology score measured 6 h after the onset of mechanical ventilation, presence of malignancy, presence of APACHE (acute physiology and chronic health evaluation) II-associated comorbidity, and the need for mechanical ventilation > or = 72 h were independent predictors of poor outcome. CONCLUSIONS: We conclude that among variables available within the first 6 h of mechanical ventilation, the presence of comorbidity and a measure of the severity of the acute illness are predictors of in-hospital mortality among patients with COPD and acute respiratory failure. The occurrence of extubation failure or the need for mechanical ventilation beyond 72 h also portends a worse prognosis. PMID- 11399714 TI - Do blood transfusions improve outcomes related to mechanical ventilation? AB - BACKGROUND: Correcting the decrease in oxygen delivery from anemia using allogeneic RBC transfusions has been hypothesized to help with increased oxygen demands during weaning from mechanical ventilation. However, it is also possible that transfusions hinder the process because RBCs may not be able to adequately increase oxygen delivery. In this study, we determined whether a liberal RBC transfusion strategy improved outcomes related to mechanical ventilation. METHODS: Seven hundred thirteen patients receiving mechanical ventilation, representing a subgroup of patients from a larger trial, were randomized to either a restrictive transfusion strategy, receiving allogeneic RBC transfusions at a hemoglobin concentration of 7.0 g/dL (and maintained between 7.0 g/dL and to 9.0 g/dL), or to a liberal transfusion strategy, receiving RBCs at 10.0 g/dL (and maintained between 10.0 g/dL and 12.0 g/dL). The larger trial was designed to evaluate transfusion practice rather than weaning per se. RESULTS: Baseline characteristics in the restrictive-strategy group (n = 357) and the liberal strategy group (n = 356) were comparable. The average durations of mechanical ventilation were 8.3 +/- 8.1 days and 8.3 +/- 8.1 days (95% confidence interval [CI] around difference, - 0.79 to 1.68; p = 0.48), while ventilator-free days were 17.5 +/- 10.9 days and 16.1 +/- 11.4 days (95% CI around difference, - 3.07 to 0.21; p = 0.09) in the restrictive-strategy group vs the liberal-strategy group, respectively. Eighty-two percent of the patients in the restrictive strategy group were considered successfully weaned and extubated for at least 24 h, compared to 78% for the liberal-strategy group (p = 0.19). The relative risk (RR) of extubation success in the restrictive-strategy group compared to the liberal-strategy group, adjusted for the confounding effects of age, APACHE (acute physiology and chronic health evaluation) II score, and comorbid illness, was 1.07 (95% CI, 0.96 to 1.26; p = 0.43). The adjusted RR of extubation success associated with restrictive transfusion in the 219 patients who received mechanical ventilation for > 7 days was 1.1 (95% CI, 0.84 to 1.45; p = 0.47). CONCLUSION: In this study, there was no evidence that a liberal RBC transfusion strategy decreased the duration of mechanical ventilation in a heterogeneous population of critically ill patients. PMID- 11399715 TI - The efficacy of postoperative incentive spirometry is influenced by the device specific imposed work of breathing. AB - STUDY OBJECTIVES: The study evaluated the impact of the additional imposed work of breathing (WBimp) generated by two different spirometers on postoperative incentive spirometry performance in patients at high risk and moderate risk for postoperative pulmonary complications (PPCs). Additionally, we investigated whether maximal inspiratory pressure (PImax) is an easy estimate of the WBimp imposed by incentive spirometers. DESIGN: Prospective, randomized, single-blind clinical trial. SETTING: ICU of a university hospital. INTERVENTIONS AND MEASUREMENTS: Thirty male patients were assigned to a group at high risk for PPCs (group A; inspiratory capacity [IC], < 1.6 L) or to a group at moderate risk for PPCs (group B; IC, 1.6 to 2.5 L) after upper-abdominal, thoracic, or two-cavity surgery. On the first or second postoperative day WBimp, IC, and PImax were recorded without spirometers (baseline) and during incentive spirometry with the Mediflo spirometer (Medimex; Hamburg, Germany) (high WBimp) and the Coach spirometer (Kendall; Neustadt, Germany) (low WBimp) using a pneumotachograph. In group A, the baseline and the ICs for both spirometers only differed slightly. In group B, the IC was significantly reduced for the Mediflo (p < 0.05), which imposed a WBimp twice as high as the Coach (p < 0.01). PImax was significantly increased for both the Mediflo and the Coach (p < 0.01). PImax was positively correlated with WBimp (r = 0.8). CONCLUSIONS: Incentive spirometers differ considerably in their additional Wbimp with a potential impact on the efficacy of postoperative incentive spirometry performance. PImax might be an easy clinical estimate for the WBimp during incentive spirometry. Incentive spirometers with low WBimp permit increased maximal sustained inspiration and, thus, enhanced incentive spirometry performance, and, therefore, it might be more suitable for use in postoperative respiratory care. PMID- 11399716 TI - Acute respiratory failure associated with pulmonary cryptococcosis in non-aids patients. AB - PURPOSE: To determine the incidence of acute respiratory failure (ARF) in non AIDS patients with pulmonary cryptococcosis (PC). DESIGN: Retrospective cohort study. SETTING: University of Pittsburgh Medical Center, Pittsburgh, PA. SUBJECTS: All patients in whom PC without HIV infection was diagnosed between February 1989 and March 1999. RESULTS: Thirty-three patients with PC were identified, and 11 of those patients (33%) developed ARF and comprised the study group. Underlying diseases included solid-organ transplant recipients (seven patients; 64%) and other underlying medical conditions (four patients; 36%). The most common symptoms were cough, shortness of breath, and temperature elevation. Extrapulmonary involvement was seen in six patients (meningitis, four patients; peritonitis, one patient; laryngeal mass, one patient). Six of the 11 patients (55%) died. CONCLUSION: ARF may develop in one third of non-AIDS patients with PC. This clinical syndrome is associated with the dissemination to extrapulmonary sites and high mortality rates. PC should be recognized as a possible cause of respiratory failure in non-AIDS patients. PMID- 11399717 TI - Pharmacologic properties of brewery dust extracts in vitro. AB - STUDY OBJECTIVES: To study the effects of extracts of brewery dust on isolated guinea pig tracheal smooth muscle in vitro. DESIGN: Parallel pharmacologic intervention on guinea pig tracheal rings that were obtained from the same animal. SETTING: Mount Sinai School of Medicine, Department of Pulmonary Medicine. MATERIAL: The isolated guinea pig tracheal tissue of 18 guinea pigs. INTERVENTIONS: Pretreatment of guinea pig rings by mediator-modifying agents before challenge with the brewery dust extracts. MEASUREMENTS AND RESULTS: The effect of brewery dust extracts on isolated guinea pig tracheal smooth muscle was studied using water-soluble extracts of dust obtained from brewery materials, including hops, barley, and brewery yeast. Dust extracts were prepared as a 1:10 (wt/vol) aqueous solution. Dose-related contractions of nonsensitized guinea pig tracheas were demonstrated using these extracts. The dust extracts contained significant quantities of bacterial components (eg, endotoxin and n-formyl methionyl-leucyl-phenylalanine), but these agents were not thought to contribute directly to the constrictor effect of the dusts. Pharmacologic studies were performed by pretreating guinea pig tracheal tissue with the following drugs known to modulate smooth muscle contraction: atropine; indomethacin; pyrilamine; LY171883; nordihydroguaiaretic acid; captopril; thiorphan; verapamil; and TMB8. The constrictor effects of the dust extracts were inhibited by a wide variety of agents, the patterns of which depended on the dust extract. Atropine consistently and strikingly reduced the contractile effects of these extracts. These observations may suggest an interaction of the extracts with parasympathetic nerves or, more directly, with muscarinic receptors. The inhibition of contraction by the blocking of other mediators was less effective and varied with the dust extract. CONCLUSIONS: We suggest that brewery dust extracts cause a dose related airway smooth muscle constriction by nonimmunologic mechanisms involving a variety of airway mediators and, possibly, cholinergic receptors. This effect is not dependent on presensitization of the guinea pigs. PMID- 11399718 TI - Methacholine-induced temporal changes in airway geometry and lung density by CT. AB - PURPOSE: Electron-beam CT (EBCT) was utilized to assess the time course of changes in airways cross-sectional area (CSA) and lung density during methacholine-induced bronchoconstriction. MATERIALS AND METHODS: EBCT scans (200 ms, 3-mm thickness, 2 mm increments) were obtained before (baseline) and 30 s, 2 min, and 4 min after bolus IV injection of methacholine to pigs receiving mechanical ventilation. A total of seven experiments were analyzed using custom made image analysis software. With each challenge, five different airways and 50 lung regions of interest were studied. RESULTS: The time course of lung density changes paralleled the time course for CSA changes. The maximal response to methacholine, measured in terms of both CSA and lung density changes, occurred 30 s after injection. Lung density changes were unaffected by reconstruction algorithm, normal (standard) or sharp (high resolution). Overall, there was increased air content in the lung during bronchoconstriction. This effect was significantly greater at the dependent lung regions. CONCLUSIONS: EBCT is an effective tool to assess temporal and regional changes in the lung during bronchoconstriction. Measurements of lung density during bronchoconstriction allow for assessment of peripheral changes that are beyond the CT spatial resolution of airways anatomy. PMID- 11399719 TI - Auditory detection of simulated crackles in breath sounds. AB - BACKGROUND: Computerized analysis of breath sounds has relied on human auditory perception as the reference standard for identifying crackles. In this study, we tested the human audibility of crackles by superimposing artificial clicks on recorded breath sounds and having physicians listen to the recordings to see if they could identify the crackles. OBJECTIVES: To establish the audibility of simulated crackles introduced in breath sounds of different intensity, to study the effects of crackle characteristics on their audibility, and to investigate crackle detection within and between observers. METHODS: Fine, medium, and coarse crackles with large and small amplitude were synthesized by computer software. Waveform parameters were based on published characteristics of lung sound crackles. The amplitude for small crackles was defined as just above the threshold of audibility for simulated crackles inserted in sound recorded during breath hold. Simulated crackles were then superimposed on breath sounds recorded at 0 L/s (breath hold), 1 L/s, and 2 L/s airflow. Five physicians listened during playback on two separate occasions to determine if crackles could be heard and to calculate the interobserver and intraobserver variations. RESULTS: Failed detection of crackles was significantly more common in the following conditions: (1) background breath sounds had higher intensity (2 L/s airflow) compared to lower intensity (1 L/s), (2) crackle type was coarse or medium compared to fine, and (3) crackle amplitude was small compared to large. Both intraobserver and interobserver agreements were high (kappa > 0.6). RELEVANCE: The validation of automated techniques for crackle detection in lung sound analysis should not rely on auscultation as the only reference. Detection of crackles is facilitated when patients take slow, deep breaths that generate little breath sounds. PMID- 11399720 TI - Pentoxifylline rescue preserves lung function in isolated canine lungs injured with phorbol myristate acetate. AB - OBJECTIVE: We hypothesized that pentoxifylline, administered after phorbol myristate acetate (PMA), would diminish the severity of lung injury. SETTING: Animal research laboratory. DESIGN: Comparative study. SUBJECTS: Mongrel dogs (n = 33). INTERVENTIONS: Baseline measurements were obtained from the isolated blood perfused dog lung lobes after 1 h of stable perfusion and ventilation. Four different measures of lung compliance were obtained along with WBC and neutrophil counts. Pulmonary vascular resistance (PVR) and capillary filtration coefficient (Kf) were calculated, and the ratio of a normalized maximal enzymatic conversion rate to the Michaelis-Menten constant (Amax/Km) was used to assess perfused capillary surface area. The control lobes (n = 8) were ventilated and perfused for an additional 40 min while the injured lobes (n = 17) received PMA (0.1 microg/mL of perfusate). The pentoxifylline-protected lobes (n = 8) were treated with pentoxifylline (1 mg/mL of perfusate) 10 min after injury with PMA. All measurements were then repeated. MEASUREMENT AND MAIN RESULTS: The three groups did not differ significantly at baseline. The control lobes remained relatively stable over time. The injured lobes demonstrated marked deterioration in compliance: 8.79 +/- 0.7 to 5.97 +/- 0.59 mL/cm H(2)O (p < 0.05) vs 10.1 +/- 1.0 to 8.07 +/- 0.72 mL/cm H(2)O and 9.6 +/- 1.1 to 9.9 +/- 0.85 mL/cm H(2)O in the control and protected lobes, respectively. Both groups receiving PMA had similar drops in WBC and neutrophil counts, but the pentoxifylline-protected lobes had preservation of all four compliance measures. PVR increased from 37.8 +/- 1.8 to 118.6 +/- 12.7 cm H(2)O/L/min (p < 0.05) in the injured lobes vs 35.4 +/- 0.5 to 36.3 +/- 2.8 cm H(2)O/L/min and 40.4 +/- 0.04 to 46.7 +/- 2.8 cm H(2)O/L/min (p < 0.05) in the control and protected lobes, respectively. Kf increased < 25% in the protected group but more than tripled in the injured group. Amax/Km dropped from 559 +/- 36 to 441 +/- 33 mL/min (p < 0.05) in the injured lobes vs 507 +/- 14 to 490 +/- 17 mL/min and 609 +/- 34 to 616 +/- 37 mL/min in the control and pentoxifylline-protected lobes, respectively. CONCLUSIONS: The use of pentoxifylline as a rescue agent prevented the PMA-induced deterioration of lung compliance, vascular integrity, and endothelial metabolic function in this acute lung injury model, despite significant pulmonary neutrophil sequestration. PMID- 11399721 TI - Talc preparations used for pleurodesis vary markedly from one preparation to another. AB - BACKGROUND: At the present time, talc is the one of the agents most commonly used for the production of a pleurodesis. However, there have been several recent reports of acute pneumonitis developing after the intrapleural administration of talc. The incidence of pneumonitis has varied markedly from center to center. OBJECTIVE: To compare the physical characteristics of talc used for the production of pleurodesis in various localities. DESIGN: Eight talc preparations (four from the United States, and one each from Brazil, France, Spain, and Taiwan) were analyzed for the distribution of the particle size and the type and amount of impurities. MEASUREMENTS: The physical characteristics of the talc specimens were determined using radiograph diffraction and scanning electron microscopy. RESULTS: The mean and median particle size varied by more than a factor of three among the eight different talc preparations. In addition, the impurities of the different talc preparations were quite varied. CONCLUSIONS: We conclude that there is marked variation in the physical characteristics of the talc preparations used intrapleurally for the production of a pleurodesis. We speculate that different incidences of acute pneumonitis at various centers after intrapleural administration of talc may be due to differences in the physical characteristics of the talc preparations used for pleurodesis. PMID- 11399722 TI - Do all patients require supplemental oxygen during flexible bronchoscopy? AB - STUDY OBJECTIVES: Some respiratory units routinely administer supplemental oxygen to all patients during flexible bronchoscopy, but other units give oxygen only to those who desaturate (arterial oxygen saturation [SaO(2)], < 90%). We performed a study to examine both the requirement for supplemental oxygen and the effect of IV midazolam therapy on oxygenation during flexible bronchoscopy for patients with a known FEV(1). DESIGN: Data on the SaO(2) of patients during flexible bronchoscopy were collected prospectively for all procedures performed in our respiratory unit for the period 1992 to 1997. RESULTS: There were 1,051 flexible bronchoscopy procedures performed in which the patient had a known FEV(1) and was not receiving supplemental oxygen before the procedure. Supplemental oxygen was commenced during or immediately after the procedure in 151 cases (14.4%), while a further 101 cases (9.6%) had momentary desaturation (ie, < 20 s) not requiring oxygen therapy. The lower the FEV(1), the greater the risk of significant desaturation and the need for supplemental oxygen (p < 0.0001) [supplemental O(2) therapy was administered in 35% of cases if FEV(1) < 1.0 L, in 14% of cases if FEV(1) was 1.0 to 1.5 L, and in 7% of cases if FEV(1) > 1.5 L]. The use of low-to moderate doses of midazolam as sedation did not affect the probability of the occurrence of significant desaturation (p = 0.204). CONCLUSIONS: This study supports guidelines that suggest that all patients should be monitored by pulse oximetry during flexible bronchoscopy. Desaturation may occur at any FEV(1) level even without sedation. The majority of our patients did not require routine oxygen supplementation, especially the group with an FEV(1) above 1 L. PMID- 11399723 TI - Successful bronchoscopic placement of tracheobronchial stents without fluoroscopy. AB - BACKGROUND: Tracheobronchial stenting is performed increasingly often. Fluoroscopic control, which leads to significant radiation exposure for patients and staff, is recommended for the placement of metal stents. METHODS: All consecutive patients referred to two airway centers in need of airway stenting who received stents (Ultraflex; Boston Scientific, Natick, MA) underwent placement using endoscopic guidance only. All data were collected in an ongoing continuous database. RESULTS: One hundred stents were placed in 96 patients for central airway obstruction, and the data were reviewed. Stents were placed in all locations within the central airways for a variety of indications but mainly for malignant obstruction. No complications occurred, and all stents were placed satisfactorily. CONCLUSION: At centers with dedicated airway teams, Ultraflex stents can be quickly and safely inserted without the need for fluoroscopy. This saves radiation exposure to patients and to staff and may lead to a more cost effective procedure. PMID- 11399724 TI - Status asthmaticus in children: a review. AB - About 10% of American children have asthma, and its prevalence, morbidity, and mortality have been increasing. Asthma is an inflammatory disease with edema, bronchial constriction, and mucous plugging. Status asthmaticus in children requires aggressive treatment with beta-agonists, anticholinergics, and corticosteroids. Intubation and mechanical ventilation should be avoided if at all possible, as the underlying dynamic hyperinflation will worsen with positive pressure ventilation. If mechanical ventilation becomes necessary, controlled hypoventilation with low tidal volume and long expiratory time may lessen the risk of barotrauma and hypotension. Unusual and nonestablished therapies for severe asthma are discussed. PMID- 11399725 TI - Top ten list in sarcoidosis. PMID- 11399726 TI - Simple treadmill score to diagnose coronary disease. AB - OBJECTIVE: Our aim was to derive and validate a simplified treadmill score for predicting the probability of angiographically confirmed coronary artery disease (CAD). BACKGROUND: The American College of Cardiology/American Heart Association guidelines for exercise testing recommend the use of multivariable equations to enhance the diagnostic characteristics of the standard treadmill test. Most of these equations use complicated statistical techniques to provide diagnostic estimates of CAD. Simplified scores derived from such equations that require physicians only to add points have been developed for pretest estimates of disease and for prognosis. However, no simplified score has been developed specifically for the diagnosis of CAD using exercise test results. METHODS: Consecutive patients referred for evaluation of chest pain who underwent standard treadmill testing followed by coronary angiography were studied. A logistic regression model was used to predict clinically significant (> or = 50% stenosis) CAD and then the variables and coefficients were used to derive a simplified score. The simplified score was calculated as follows: (6 x maximal heart rate code) + (5 x ST-segment depression code) + (4 x age code) + angina pectoris code + hypercholesterolemia code + diabetes code + treadmill angina index code. The simplified score had a range from 6 to 95, with < 40 designated as low probability, between 40 and 60 was intermediate probability, and > 60 was high probability for CAD. RESULTS: A total of 1,282 male patients without a prior myocardial infarction underwent exercise treadmill testing and coronary angiography in the derivation group, and there were 476 male patients in the validation group from another institution. The area under the receiver operating characteristic curve (+/- SE) for the ST-segment response alone was 0.67 as compared to 0.79 +/- 0.01 for the diagnostic score (p > 0.001). The prevalence of significant disease for the men was 27% in the low-probability group, 62% in the intermediate-probability group, and 92% in the high-probability group, which was similar to the prevalence in the validation group, with 22%, 58%, and 92% in low , intermediate-, and high-probability groups, respectively. The low-probability group had < 4% prevalence of severe disease. In both populations, 7 more patients out of 100 were correctly classified than with the use of ST-segment criteria. When used as a clinical management strategy, the score has a sensitivity of 88% and a specificity of 96%. CONCLUSION: This simplified exercise score that estimates the probability of CAD can be easily applied without a calculator and is a useful and valid tool that can help physicians manage patients presenting with chest pain. PMID- 11399727 TI - Lung cancer resection or aortic graft replacement with simultaneous myocardial revascularization without cardiopulmonary bypass. AB - BACKGROUND: The concomitant occurrence of lung cancer or other thoracic problems requiring surgical treatment in patients with significant coronary artery disease is uncommon. METHODS: Three patients underwent revascularization of the anterior descending artery, without cardiopulmonary bypass, with simultaneous pulmonary lobectomy (two patients) or replacement of an obstructed descending aortic graft (one patient). RESULTS: Postoperative ventilation time was < 3 h, and no morbidity related to the combined procedure occurred during midterm follow-up. CONCLUSIONS: This one-stage approach allowed the immediate solution of two intrathoracic comorbidities, reducing expenses and suffering to the patients and minimizing the risk of bleeding or tumor dissemination secondary to extracorporeal circulation-induced coagulopathy and immunosuppression. PMID- 11399728 TI - Motivating factors in futile clinical interventions. AB - With modern medical technology, it is now possible to sustain life for prolonged periods in critically ill patients, even when there is no reasonable hope of improvement or achieving the goals of therapy. Such futile and medically inappropriate interventions may violate both the ethical and medical precepts generally accepted by patients, families, and physicians. In this study, we sought to determine who was primarily responsible for such interventions, the nature of their motivation, and the role of a timely bioethical consultation. In a retrospective review, we identified 100 patients of 331 bioethical consultations who had futile or medically inappropriate therapy. The average age of patients was 73.5 +/- 32 years (mean +/- 2 SD) with 57% being male. Fifty seven percent of the patients were admitted to the hospital with a degenerative disorder, 21% with an inflammatory disorder, and 16% with a neoplastic disorder. The family was responsible for futile treatment in 62% of cases, the physician in 37% of cases, and a conservator in one case. Unreasonable expectation for improvement was the most common underlying factor. Family dissent was involved in 7 of 62 cases motivated by family, but never when physicians were primarily responsible. Liability issues motivated physicians in 12 of 37 cases where they were responsible but in only 1 of 62 cases when the family was (chi(2) 5 degrees of freedom = 26.7, p < 0.001). When the bioethics consultation resulted in cessation of the therapy, patients died in a median of 2 days as opposed to 16 days if therapy continued (p < 0.001). PMID- 11399729 TI - A 51-year-old man with a complicated medical history and abnormal chest radiographic finding. PMID- 11399730 TI - A patient with palpitations and bilateral pulmonary infiltrates. PMID- 11399731 TI - Respiratory arrest in the eye clinic. PMID- 11399732 TI - Tracheal injury caused by ingested paraquat. AB - Paraquat is a potent herbicide of lethal toxicity. Its injury mechanism is attributed to the generation of very-reactive oxygen species, such as superoxide radicals, through which multiple injuries are produced on mucosa, although there have been no reports of injuries on the trachea. We describe a case of fatal paraquat poisoning with tracheal injuries, where the clinical debut was acute respiratory insufficiency and a spontaneous pneumothorax. PMID- 11399733 TI - Severe pectus excavatum associated with cor pulmonale and chronic respiratory acidosis in a young woman. AB - Pectus excavatum has never been reported to cause hypercapnic respiratory failure. In this report, we describe the first such case in a young woman with severe pectus excavatum who presented with chronic respiratory acidosis, pulmonary hypertension, and chronic cor pulmonale. An extensive diagnostic workup failed to uncover any other cause of respiratory acidosis, which led us to conclude that the severe chest wall deformity and the resulting severe restrictive defect were responsible for the development of chronic respiratory acidosis and cor pulmonale. PMID- 11399734 TI - Severe immune hemolytic anemia in disseminated tuberculosis with response to antituberculosis therapy. AB - Severe hemolytic anemia in patients with disseminated tuberculosis is exceedingly rare. We report an episode of Coombs'-positive hemolytic anemia in a previously healthy young man with miliary tuberculosis, resulting in a hemoglobin level of 5 g/dL and an undetectable haptoglobin level. The patient responded well to treatment with antituberculosis drugs, and the results of the direct Coombs' test became negative without the need of blood transfusion or steroid therapy. PMID- 11399735 TI - Ticlopidine-induced interstitial pulmonary disease: a case report. AB - We report a case of interstitial pulmonary disease that occurred together with lymphocytic colitis during treatment with ticlopidine. The drug was prescribed for transient ischemic cerebrovascular accidents. Ticlopidine treatment was stopped, and a prolonged course of prednisone was necessary to treat the pulmonary and intestinal symptoms. So far, few cases of pulmonary side effects caused by ticlopidine have been reported. This case is unique in that interstitial lung disease evolved in parallel with colitis and caused severe hypoxemia. Special care should be taken when pulmonary symptoms appear in association with ticlopidine treatment. PMID- 11399736 TI - Bilevel nasal positive airway pressure and ballooning of the stomach. AB - We describe a case of severe gastric insufflation in a patient with amyotrophic lateral sclerosis who was receiving bilevel nasal positive airway pressure (BNPAP) ventilation (BiPAP; Respironics; Murrysville, PA). The injection of inspiratory flow into the esophagus, aerophagia, and air trapping below the gastroesophageal junction after a meal are probably the major causes. We suggest that BNPAP ventilation can be a cause of serious gastric insufflation in a patient who lies supine, especially after a meal, and attention should be paid to avoiding this complication by having the patient sit up for about half an hour after a meal. PMID- 11399737 TI - Utility of Wang needle aspiration in the diagnosis of actinomycosis. AB - An 85-year-old man had a 4-year history of recurrent pneumonia with a persistent pleural effusion. He underwent repeated bronchoscopy that revealed a right bronchus intermedius mass, but bronchial washes and biopsies remained nondiagnostic. A repeat bronchoscopy was performed, and a Wang needle aspiration of the mass was obtained that showed sulfur granules, diagnosing actinomycosis. The patient was started on appropriate antibiotic therapy. Actinomycosis must be considered in a patient with recurrent pneumonia and an endobronchial mass. Wang needle aspiration via bronchoscopy may be an important diagnostic tool. PMID- 11399738 TI - The use of endoscopic argon plasma coagulation in airway complications after solid organ transplantation. AB - The objective of the study was to describe a safe and effective treatment option for endobronchial complications after solid organ transplantation. A retrospective analysis was performed in a tertiary-care university hospital. The use of bronchoscopic argon plasma coagulation (APC) for the treatment of endobronchial lesions was studied in five solid organ transplant recipients. Four patients presented with variable degrees of endobronchial obstruction, and one patient presented with massive hemoptysis. Two of the patients with endobronchial obstruction were double lung transplant recipients who developed anastomotic strictures. The strictures were opened with endobronchial stents but became obstructed again by inflammatory granulation tissue overgrowth through the stent mesh. APC was used to maintain airway patency. One kidney transplant recipient developed pulmonary zygomycosis with secondary obstruction of the left main bronchus because of granulation tissue growth through endobronchial stents. Airway patency was reestablished with several treatments with APC. Another kidney transplant recipient developed subglottic and tracheal papillomatosis that was effectively removed with APC. A heart transplant recipient was referred with recurrent massive hemoptysis refractory to bronchial artery embolization. The bleeding was caused by hemorrhagic polypoid lesions, which were completely ablated by APC. Bronchoscopic use of the argon plasma coagulator is a safe and simple technique that can be used effectively to treat endobronchial pathology in solid organ transplant patients. PMID- 11399739 TI - Blebs and/or bullae are of no importance and have no predictive value for recurrences in patients with primary spontaneous pneumothorax. PMID- 11399740 TI - Free range chicken soup. PMID- 11399741 TI - High-frequency ventilation and the prevention of ventilator-associated lung injury. PMID- 11399742 TI - Lipid-lowering agents and artery endothelial function. PMID- 11399743 TI - Pulmonary arterial hypertension and thyroid disease. PMID- 11399744 TI - A survey of the use of herbal agents among Philadelphia Veterans Affairs Medical Center pulmonary outpatients. PMID- 11399745 TI - Elastic recoil of lung and lung reduction surgery. PMID- 11399746 TI - Estrogen and spermatogenesis. AB - Although it has been known for many years that estrogen administration has deleterious effects on male fertility, data from transgenic mice deficient in estrogen receptors or aromatase point to an essential physiological role for estrogen in male fertility. This review summarizes the current knowledge on the localization of estrogen receptors and aromatase in the testis in an effort to understand the likely sites of estrogen action. The review also discusses the many studies that have used models employing the administration of estrogenic substances to show that male fertility is responsive to estrogen, thus providing a mechanism by which inappropriate exposure to estrogenic substances may cause adverse effects on spermatogenesis and male fertility. The reproductive phenotypes of mice deficient in estrogen receptors alpha and/or beta and aromatase are also compared to evaluate the physiological role of estrogen in male fertility. The review focuses on the effects of estrogen administration or deprivation, primarily in rodents, on the hypothalamo-pituitary-testis axis, testicular function (including Leydig cell, Sertoli cell, and germ cell development and function), and in the development and function of the efferent ductules and epididymis. The requirement for estrogen in normal male sexual behavior is also reviewed, along with the somewhat limited data on the fertility of men who lack either the capacity to produce or respond to estrogen. This review highlights the ability of exogenous estrogen exposure to perturb spermatogenesis and male fertility, as well as the emerging physiological role of estrogens in male fertility, suggesting that, in this local context, estrogenic substances should also be considered "male hormones." PMID- 11399747 TI - Environmental signaling: what embryos and evolution teach us about endocrine disrupting chemicals. AB - The term "endocrine disrupting chemicals" is commonly used to describe environmental agents that alter the endocrine system. Laboratories working in this emerging field-environmental endocrine research-have looked at chemicals that mimic or block endogenous vertebrate steroid hormones by interacting with the hormone's receptor. Environmental chemicals known to do this do so most often with receptors derived from the steroid/thyroid/retinoid gene family. They include ubiquitous and persistent organochlorines, as well as plasticizers, pharmaceuticals, and natural hormones. These chemicals function as estrogens, antiestrogens, and antiandrogens but have few, if any, structural similarities. Therefore, receptor-based or functional assays have the best chance of detecting putative biological activity of environmental chemicals. Three nuclear estrogen receptor forms-alpha, beta, and gamma-as well as multiple membrane forms and a possible mitochondrial form have been reported, suggesting a previously unknown diversity of signaling pathways available to estrogenic chemicals. Examples of environmental or ambient estrogenization occur in laboratory experiments, zoo animals, domestic animals, wildlife, and humans. Environmentally estrogenized phenotypes may differ depending upon the time of exposure-i.e., whether the exposure occurred at a developmental (organizational and irreversible) or postdevelopmental (activational and reversible) stage. The term "estrogen" must be defined in each case, since steroidal estrogens differ among themselves and from synthetic or plant-derived chemicals. An "estrogen-like function" seems to be an evolutionarily ancient signal that has been retained in a number of chemicals, some of which are vertebrate hormones. Signaling, required for symbiosis between plants and bacteria, may be viewed, therefore, as an early example of hormone cross-talk. Developmental feminization at the structural or functional level is an emerging theme in species exposed, during embryonic or fetal life, to estrogenic compounds. Human experience as well as studies in experimental animals with the potent estrogen diethylstilbestrol provide informative models. Advances in the molecular genetics of sex differentiation in vertebrates facilitate mechanistic understanding. Experiments addressing the concept of gene imprinting or induction of epigenetic memory by estrogen or other hormones suggest a link to persistent, heritable phenotypic changes seen after developmental estrogenization, independent of mutagenesis. Environmental endocrine science provides a new context in which to examine the informational content of ecosystem-wide communication networks. As common features come to light, this research may allow us to predict environmentally induced alterations in internal signaling systems of vertebrates and some invertebrates and eventually to explicate environmental contributions to human reproductive and developmental health. PMID- 11399748 TI - Male sexual function and its disorders: physiology, pathophysiology, clinical investigation, and treatment. AB - This review is designed to help the reproductive endocrinologist integrate his or her professional activity with those of other disciplines including urology, radiology, neurology, and psychology in order to successfully manage all of the inseparable aspects of male sexual and reproductive functioning. Significant advances in the field of male sexual physiology and pathophysiology and new methods of investigation and treatment of male sexual disorders are outlined. The review synthesizes available data on the following: norms of sexual organs, aging and sexuality, role of central and peripheral neurochemicals in each stage of the sexual cycle, role of corporeal smooth muscles in the hemodynamic control of erection and detumescence, influence of psychological factors, drugs, and disease on all aspects of sexual functioning, and use of nocturnal penile tumescence monitoring, imaging investigations, and neurophysiologic studies in the diagnostic workup of males with sexual dysfunction. Clinical algorithms are presented where appropriate. Extensive discussions on newly developed strategies in psychological and behavioral counseling, drug therapy, tissue engineering, nonsurgical devices, and surgical treatments for all forms of sexual disorders are also provided. Lastly, the effect of sexual dysfunction and its treatment on quality of life in affected men is addressed, along with recommendations for future research endeavors. PMID- 11399749 TI - Are estrogens of import to primate/human ovarian folliculogenesis? AB - The notion that estrogens play a meaningful role in ovarian folliculogenesis stems from a large body of in vitro and in vivo experiments carried out in certain rodent models, (e.g., rats) wherein the stimulatory role of estrogen on granulosa cell growth and differentiation is undisputed. However, evidence derived from these polyovulatory species may not be readily generalizable to the monoovulatory subhuman primates, let alone the human. Only recently, significant observations on the ovarian role(s) of estrogen have been reported for the primate/human. It is thus the objective of this communication to review the evidence for and against a role for estrogens in primate/human ovarian follicular development with an emphasis toward the application of the concepts so developed to contemporary reproductive physiology and to the practice of reproductive medicine. The role(s) of estrogens will be examined not only by analyzing the physiological evidence to the effect that these hormones control ovarian function and follicular growth, but also by summarizing the molecular evidence for the existence and distribution of the cognate receptors. PMID- 11399750 TI - Interaction between aldolase and vacuolar H+-ATPase: evidence for direct coupling of glycolysis to the ATP-hydrolyzing proton pump. AB - Vacuolar H(+)-ATPases (V-ATPases) are essential for acidification of intracellular compartments and for proton secretion from the plasma membrane in kidney epithelial cells and osteoclasts. The cellular proteins that regulate V ATPases remain largely unknown. A screen for proteins that bind the V-ATPase E subunit using the yeast two-hybrid assay identified the cDNA clone coded for aldolase, an enzyme of the glycolytic pathway. The interaction between E subunit and aldolase was confirmed in vitro by precipitation assays using E subunit glutathione S-transferase chimeric fusion proteins and metabolically labeled aldolase. Aldolase was isolated associated with intact V-ATPase from bovine kidney microsomes and osteoclast-containing mouse marrow cultures in co immunoprecipitation studies performed using an anti-E subunit monoclonal antibody. The interaction was not affected by incubation with aldolase substrates or products. In immunocytochemical assays, aldolase was found to colocalize with V-ATPase in the renal proximal tubule. In osteoclasts, the aldolase-V-ATPase complex appeared to undergo a subcellular redistribution from perinuclear compartments to the ruffled membranes following activation of resorption. In yeast cells deficient in aldolase, the peripheral V(1) domain of V-ATPase was found to dissociate from the integral membrane V(0) domain, indicating direct coupling of glycolysis to the proton pump. The direct binding interaction between V-ATPase and aldolase may be a new mechanism for the regulation of the V-ATPase and may underlie the proximal tubule acidification defect in hereditary fructose intolerance. PMID- 11399751 TI - Calcium-stimulated autophosphorylation site of plant chimeric calcium/calmodulin dependent protein kinase. AB - The existence of two molecular switches regulating plant chimeric Ca(2+)/calmodulin-dependent protein kinase (CCaMK), namely the C-terminal visinin like domain acting as Ca(2+)-sensitive molecular switch and calmodulin binding domain acting as Ca(2+)-stimulated autophosphorylation-sensitive molecular switch, has been described (Sathyanarayanan, P. V., Cremo, C. R., and Poovaiah, B. W. (2000) J. Biol. Chem. 275, 30417-30422). Here we report the identification of Ca(2+)-stimulated autophosphorylation site of CCaMK by matrix-assisted laser desorption ionization time of flight-mass spectrometry. Thr(267) was confirmed as the Ca(2+)-stimulated autophosphorylation site by post-source decay experiments and by site-directed mutagenesis. The purified T267A mutant form of CCaMK did not show Ca(2+)-stimulated autophosphorylation, autophosphorylation-dependent variable calmodulin affinity, or Ca(2+)/calmodulin stimulation of kinase activity. Sequence comparison of CCaMK from monocotyledonous plant (lily) and dicotyledonous plant (tobacco) suggests that the autophosphorylation site is conserved. This is the first identification of a phosphorylation site specifically responding to activation by second messenger system (Ca(2+) messenger system) in plants. Homology modeling of the kinase and calmodulin binding domain of CCaMK with the crystal structure of calcium/calmodulin dependent protein kinase 1 suggests that the Ca(2+)-stimulated autophosphorylation site is located on the surface of the kinase and far from the catalytic site. Analysis of Ca(2+)-stimulated autophosphorylation with increasing concentration of CCaMK indicates the possibility that the Ca(2+)-stimulated phosphorylation occurs by an intermolecular mechanism. PMID- 11399752 TI - Sequential activation of protein kinase C (PKC)-alpha and PKC-epsilon contributes to sustained Raf/ERK1/2 activation in endothelial cells under mechanical strain. AB - Endothelial cells (ECs) are constantly subjected to hemodynamic forces including cyclic pressure-induced strain. The role of protein kinase C (PKC) in cyclic strain-treated ECs was studied. PKC activities were induced as cyclic strain was initiated. Cyclic strain to ECs caused activation of PKC-alpha and -epsilon. The translocation of PKC-alpha and -epsilon but not PKC-beta from the cytosolic to membrane fraction was observed. An early transient activation of PKC-alpha versus a late but sustained activation of PKC-epsilon was shown after the onset of cyclic strain. Consistently, a sequential association of PKC-alpha and -epsilon with the signaling molecule Raf-1 was shown. ECs treated with a PKC inhibitor (calphostin C) abolished the cyclic strain-induced Raf-1 activation. ECs under cyclic strain induced a sustained activation of extracellular signal-regulated protein kinases (ERK1/2), which was inhibited by treating ECs with calphostin C. ECs treated with a specific Ca(2+)-dependent PKC inhibitor (Go 6976) showed an inhibition in the early phase of ERK1/2 activation but not in the late and sustained phase. ECs transfected with the antisense to PKC-alpha, the antisense to PKC-epsilon, or the inhibition peptide to PKC-epsilon reduced strain-induced ERK1/2 phosphorylation in a temporal manner. PKC-alpha mediated mainly the early ERK1/2 activation, whereas PKC-epsilon was involved in the sustained ERK1/2 activation. Strained ECs increased transcriptional activity of Elk1 (an ERK1/2 substrate). ECs transfected with the antisense to each PKC isoform reduced Elk1 and monocyte chemotactic protein-1 promotor activity. Our findings conclude that a sequential activation of PKC isoform (alpha and epsilon) contribute to Raf/ERK1/2 activation, and PKC-epsilon appears to play a key role in endothelial adaptation to hemodynamic environment. PMID- 11399753 TI - Conformationally sensitive residues in transmembrane domain 9 of the Na+/dicarboxylate co-transporter. AB - The Na(+)/dicarboxylate co-transporter, NaDC-1, couples the transport of sodium and Krebs cycle intermediates, such as succinate and citrate. Previous studies identified two functionally important amino acids, Glu-475 and Cys-476, located in transmembrane domain (TMD) 9 of NaDC-1. In the present study, each amino acid in TMD-9 was mutated to cysteine, one at a time, and the accessibility of the membrane-impermeant reagent [2-(trimethylammonium)ethyl]methanethiosulfonate (MTSET) to the replacement cysteines was determined. Cysteine substitution was tolerated at all but five of the sites: the A461C mutant was not present at the plasma membrane, whereas the F473C, T474C, E475C, and N479C mutants were inactive proteins located on the plasma membrane. Cysteine substitution of four residues found near the extracellular surface of TMD-9 (Ser-478, Ala-480, Ala-481, and Thr 482) resulted in proteins that were sensitive to inhibition by MTSET. The accessibility of MTSET to the four substituted cysteines was highest in the presence of the transported cations, sodium or lithium, and low in choline. The four mutants also exhibited substrate protection of MTSET accessibility. The MTSET accessibility to S478C, A481C, and A480C was independent of voltage. In contrast, T482C was more accessible to MTSET in choline buffer at negative holding potentials, but there was no effect of voltage in sodium buffer. In conclusion, TMD-9 may be involved in transducing conformational changes between the cation-binding sites and the substrate-binding site in NaDC-1, and it may also form part of the translocation pathway through the transporter. PMID- 11399754 TI - Identification and molecular cloning of a novel brain-specific receptor protein that binds to brain injury-derived neurotrophic peptide. Possible role for neuronal survival. AB - Brain injury-derived neurotrophic peptide (BINP) is a synthetic 13-mer peptide that supports neuronal survival and protects hippocampal neurons in primary cultures from cell death caused by glutamate. We have developed a monoclonal antibody named mAb 6A22 against the 40-kDa BINP-binding protein, p40BBP. mAb 6A22 inhibits binding between BINP and rat brain synaptosomes and abolishes the protective effect of BINP. The antigen of mAb 6A22 should be the BINP-binding protein that mediates the neuroprotective action of BINP. Using an expression cloning approach with mAb 6A22, we isolated a cDNA encoding a novel receptor protein that shows binding activity of BINP. COS7 cells transfected with the cloned cDNA show binding of BINP and cell surfaces that are stained by 6A22. The mRNA for p40BBP is specific for the rat brain and is increased after birth. From immunohistochemical studies using mAb 6A22, p40BBP increased after kainic acid treatment in rat hippocampal neurons. PMID- 11399755 TI - Characterization of Sptrx, a novel member of the thioredoxin family specifically expressed in human spermatozoa. AB - Thioredoxins (Trx) are small ubiquitous proteins that participate in different cellular processes via redox-mediated reactions. We report here the identification and characterization of a novel member of the thioredoxin family in humans, named Sptrx (sperm-specific trx), the first with a tissue-specific distribution, located exclusively in spermatozoa. Sptrx open reading frame encodes for a protein of 486 amino acids composed of two clear domains: an N terminal domain consisting of 23 highly conserved repetitions of a 15-residue motif and a C-terminal domain typical of thioredoxins. Northern analysis and in situ hybridization shows that Sptrx mRNA is only expressed in human testis, specifically in round and elongating spermatids. Immunostaining of human testis sections identified Sptrx protein in spermatids, while immunofluorescence and immunogold electron microscopy analysis demonstrated Sptrx localization in the cytoplasmic droplet of ejaculated sperm. Sptrx appears to have a multimeric structure in native conditions and is able to reduce insulin disulfide bonds in the presence of NADPH and thioredoxin reductase. During mammalian spermiogenesis in testis seminiferous tubules and later maturation in epididymis, extensive reorganization of disulfide bonds is required to stabilize cytoskeletal sperm structures. However, the molecular mechanisms that control these processes are not known. The identification of Sptrx with an expression pattern restricted to the postmeiotic phase of spermatogenesis, when the sperm tail is organized, suggests that Sptrx might be an important factor in regulating critical steps of human spermiogenesis. PMID- 11399756 TI - Akt is activated in response to an apoptotic signal. AB - Akt is a serine-threonine kinase known to exert antiapoptotic effects through several downstream targets. Akt is cleaved during mitochondrial-mediated apoptosis in a caspase-dependent manner. The reason for this is not clear, however, because Akt has not been demonstrated to be activated in response to mitochondrial apoptotic stimuli. Accordingly, we explored whether the well described mitochondrial apoptotic stimuli staurosporine (STS) and etoposide activate Akt and whether such activation impacts apoptosis. Both STS and etoposide activated Akt in NIH 3T3 cells, maximally at 8 and 2 h, respectively, preceding the onset of apoptosis and poly(ADP-ribose) polymerase cleavage. The overexpression of Akt delayed STS-induced apoptosis with an even more pronounced delay observed with overexpression of constitutively active Akt. Akt activation by proapoptotic stimuli lay upstream of mitochondria, because neither caspase inhibitors nor overexpression of Bcl-2 or Bcl-x(L) could prevent it. Activation depended on phosphatidylinositol 3-kinase activity, however. Conversely, inhibition of phosphatidylinositol 3-kinase with wortmannin sensitized cells to apoptosis initiated by STS. These data demonstrate that mitochondrial apoptotic stimuli also activate Akt and such activation modulates apoptosis in this setting. PMID- 11399757 TI - Loss of clumping factor B fibrinogen binding activity by Staphylococcus aureus involves cessation of transcription, shedding and cleavage by metalloprotease. AB - The fibrinogen-binding protein clumping factor B (ClfB) of Staphylococcus aureus is present on the surface of cells from the early exponential phase of growth in greater amounts than on cells from late exponential phase and is barely detectable on cells from stationary phase. Expression of a clfB-lacZ fusion indicated that transcription stopped before the end of exponential phase. Mutations in the global regulators agr and sar had no effect on clfB transcription. The loss of ClfB protein from cells in stationary phase was due to expression ending before cells stopped growing, combined with shedding of some of the protein into the growth medium and dilution of those molecules remaining on the cell surface during the two to three cell division events leading to stationary phase. Two forms of the protein occurred on the cell surface, the smaller of which was generated by loss of a domain from the N terminus. The proportion of the smaller form increased as the cultures grew. The metalloprotease aureolysin was shown to be responsible for cleavage of ClfB. Cleavage was inhibited by EDTA and o-phenanthroline and did not occur in an aureolysin-deficient mutant. Purified aureolysin promoted cleavage of cell surface-located ClfB as well as the recombinant A domain of ClfB. Cleavage was detected at two sites, one located between residues Ser(197) and Leu(198) and the other between Ala(199) and Val(200). The truncated form of ClfB did not bind fibrinogen. PMID- 11399758 TI - Roles of the basic helix-loop-helix genes Hes1 and Hes5 in expansion of neural stem cells of the developing brain. AB - Neural stem cells, which differentiate into neurons and glia, are present in the ventricular zone of the embryonal brain. The precise mechanism by which neural stem cells are maintained during embryogenesis remains to be determined. Here, we found that transient misexpression of the basic helix-loop-helix genes Hes1 and Hes5 keeps embryonal telencephalic cells undifferentiated although they have been shown to induce gliogenesis in the retina. These telencephalic cells later differentiate into neurons and astroglia when Hes expression is down-regulated, suggesting that Hes1- and Hes5- expressing cells are maintained as neural stem cells during embryogenesis. Conversely, in the absence of Hes1 and Hes5, neural stem cells are not properly maintained, generating fewer and smaller neurospheres than the wild type. These results indicate that Hes1 and Hes5 play an important role in the maintenance of neural stem cells but not in gliogenesis in the embryonal telencephalon. PMID- 11399759 TI - Biochemical studies of Zmpste24-deficient mice. AB - Genetic studies in Saccharomyces cerevisiae identified two genes, STE24 and RCE1, involved in cleaving the three carboxyl-terminal amino acids from isoprenylated proteins that terminate with a CAAX sequence motif. Ste24p cleaves the carboxyl terminal "-AAX" from the yeast mating pheromone a-factor, whereas Rce1p cleaves the -AAX from both a-factor and Ras2p. Ste24p also cleaves the amino terminus of a-factor. The mouse genome contains orthologues for both yeast RCE1 and STE24. We previously demonstrated, with a gene-knockout experiment, that mouse Rce1 is essential for development and that Rce1 is entirely responsible for the carboxyl terminal proteolytic processing of the mouse Ras proteins. In this study, we cloned mouse Zmpste24, the orthologue for yeast STE24 and showed that it could promote a-factor production when expressed in yeast. Then, to assess the importance of Zmpste24 in development, we generated Zmpste24-deficient mice. Unlike the Rce1 knockout mice, Zmpste24-deficient mice survived development and were fertile. Since no natural substrates for mammalian Zmpste24 have been identified, yeast a-factor was used as a surrogate substrate to investigate the biochemical activities in membranes from the cells and tissues of Zmpste24 deficient mice. We demonstrate that Zmpste24-deficient mouse membranes, like Ste24p-deficient yeast membranes, have diminished CAAX proteolytic activity and lack the ability to cleave the amino terminus of the a-factor precursor. Thus, both enzymatic activities of yeast Ste24p are conserved in mouse Zmpste24, but these enzymatic activities are not essential for mouse development or for fertility. PMID- 11399760 TI - Human Ca(2+) receptor extracellular domain. Analysis of function of lobe I loop deletion mutants. AB - The G protein-coupled Ca(2+) receptor (CaR) possesses an approximately 600 residue extracellular domain involved in ligand binding and receptor activation. Based on an alignment of the amino acid sequence of the CaR with that of bacterial periplasmic-binding proteins, the first approximately 530 residues of the extracellular domain are believed to form a domain resembling a bilobed Venus's flytrap (VFT). Four insertions in the CaR sequence that do not align with those of bacterial periplasmic-binding proteins correspond to four loops within lobe I of the VFT. We constructed a series of deletion mutants of these four loops and tested their ability to form fully processed CaR as well as their ability to be activated by Ca(2+). As many as 21 residues (365) of loop III could be deleted without impairing receptor expression or activation. Deletion of portions of either loops I (50) or IV (438) did not impair receptor expression but significantly reduced Ca(2+) activation. Deletion of the entire loop II (117) abolished receptor expression and function, but the replacement of even a single residue within this deletion mutant led to expression of a monomeric form of the receptor showing increased Ca(2+) sensitivity but reduced maximal activation. Our results reveal that certain residues within loops I and IV are dispensable in formation of the VFT domain but are critical for Ca(2+) activation of the receptor. In contrast, the residues in loop II are critical for maintaining the inactive state of the CaR. We discuss these results in light of the recently defined crystal structure of the homologous domain of the type 1 metabotropic glutamate receptor. PMID- 11399761 TI - Biochemical characterization of uracil processing activities in the hyperthermophilic archaeon Pyrobaculum aerophilum. AB - Deamination of cytosine to uracil and 5-methylcytosine to thymine represents a major mutagenic threat particularly at high temperatures. In double-stranded DNA, these spontaneous hydrolytic reactions give rise to G.U and G.T mispairs, respectively, that must be restored to G.C pairs prior to the next round of DNA replication; if left unrepaired, 50% of progeny DNA would acquire G.C --> A.T transition mutations. The genome of the hyperthermophilic archaeon Pyrobaculum aerophilum has been recently shown to encode a protein, Pa-MIG, a member of the endonuclease III family, capable of processing both G.U and G.T mispairs. We now show that this latter activity is undetectable in crude extracts of P. aerophilum. However, uracil residues in G.U mispairs, in A.U pairs, and in single stranded DNA were efficiently removed in these extracts. These activities were assigned to a approximately 22-kDa polypeptide named Pa-UDG (P. aerophilum uracil DNA glycosylase). The recombinant Pa-UDG protein is highly thermostable and displays a considerable degree of homology to the recently described uracil-DNA glycosylases from Archaeoglobus fulgidus and Thermotoga maritima. Interestingly, neither Pa-MIG nor Pa-UDG was inhibited by UGI, a generic inhibitor of the UNG family of uracil glycosylases. Yet a small fraction of the total uracil processing activity present in crude extracts of P. aerophilum was inhibited by this peptide. This implies that the hyperthermophilic archaeon possesses at least a three-pronged defense against the mutagenic threat of hydrolytic deamination of cytosines in its genomic DNA. PMID- 11399762 TI - Peroxisome proliferator-activated receptor gamma and chicken ovalbumin upstream promoter transcription factor II negatively regulate the phosphoenolpyruvate carboxykinase promoter via a common element. AB - A heterodimer of peroxisome proliferator-activated receptor gamma (PPARgamma) and retinoid X receptor (RXR) is required for adipocyte differentiation. The gene encoding cytosolic phosphoenolpyruvate carboxykinase (PEPCK) is a PPARgamma/RXR target gene in adipose tissue. Of the two PPARgamma response elements, gAF1/PCK1 and PCK2, only PCK2 is required for PEPCK expression and responsiveness to the PPARgamma agonist, rosiglitazone, in adipose tissue even though both elements bind PPARgamma/RXR in vitro. In contrast, gAF1/PCK1 is essential for glucocorticoid inhibition of PPARgamma-induced PEPCK gene expression in adipocytes. We report that chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) is the predominant nuclear receptor bound to gAF1/PCK1 in preadipocytes. COUP-TFII declines during adipogenesis in reciprocal fashion to PPARgamma. In transiently transfected fibroblasts COUP-TFII acts at gAF1/PCK1 to inhibit PPARgamma/RXR activation via PCK2. In contrast COUP-TFs are transcriptional activators of PEPCK in hepatocytes. PPARgamma/RXR occupies gAF1/PCK1 in adipocytes, and mutation of gAF1/PCK1 enhances PEPCK promoter transactivation by PPARgamma/RXR in fibroblasts, suggesting that this element is also a negative PPARgamma response element. These results indicate that gAF1/PCK1 is a pleiotropic element through which COUP-TFII inhibits premature PEPCK expression, and perhaps adipogenesis in general, and PPARgamma/RXR uses this same element in adipocytes to participate in PEPCK modulation by glucocorticoids. PMID- 11399763 TI - Extracellular matrix-derived peptide binds to alpha(v)beta(3) integrin and inhibits angiogenesis. AB - Angiogenesis is associated with several pathological disorders as well as with normal physiological maintenance. Components of vascular basement membrane are speculated to regulate angiogenesis in both positive and negative manner. Recently, we reported that tumstatin (the NC1 domain of alpha 3 chain of type IV collagen) and its deletion mutant tum-5 possess anti-angiogenic activity. In the present study, we confirm that the anti-angiogenic activity of tumstatin and tum 5 is independent of disulfide bond requirement. This property of tum-5 allowed us to use overlapping synthetic peptide strategy to identify peptide sequence(s) which possess anti-angiogenic activity. Among these peptides, only the T3 peptide (69-88 amino acids) and T7 peptide (74-98 amino acids) inhibited proliferation and induced apoptosis specifically in endothelial cells. The peptides, similar to tumstatin and the tum-5 domain, bind and function via alpha(v)beta(3) in an RGD independent manner. Restoration of a disulfide bond between two cysteines within the peptide did not alter the anti-angiogenic activity. Additionally, these studies show that tumstatin peptides can inhibit proliferation of endothelial cells in the presence of vitronectin, fibronectin, and collagen I. Anti angiogenic effect of the peptides was further confirmed in vivo using a Matrigel plug assay in C57BL/6 mice. Collectively, these experiments suggest that the anti angiogenic activity of tumstatin is localized to a 25-amino acid region of tumstatin and it is independent of disulfide bond linkage. Structural features and potency of the tumstatin peptide make it highly feasible as a potential anti cancer drug. PMID- 11399764 TI - Proteolytic processing and primary structure of Plasmodium falciparum apical membrane antigen-1. AB - Plasmodium falciparum apical membrane antigen-1 (PfAMA-1) is a malaria merozoite integral membrane protein that plays an essential but poorly understood role in invasion of host erythrocytes. The PfAMA-1 ectodomain comprises three disulfide constrained domains, the first of which (domain I) is preceded by an N-terminal prosequence. PfAMA-1 is initially routed to secretory organelles at the apical end of the merozoite, where the 83-kDa precursor (PfAMA-1(83)) is converted to a 66-kDa form (PfAMA-1(66)). At about the time of erythrocyte invasion, PfAMA-1(66) selectively translocates onto the merozoite surface. Here we use direct microsequencing and mass spectrometric peptide mass fingerprinting to characterize in detail the primary structure and proteolytic processing of PfAMA 1. We have determined the site at which processing takes place to convert PfAMA 1(83) to PfAMA-1(66) and have shown that both species possess a completely intact and unmodified transmembrane and cytoplasmic domain. Following relocation to the merozoite surface, PfAMA-1(66) is further proteolytically cleaved at one of two alternative sites, either between domains II and III, or at a membrane-proximal site following domain III. As a result, the bulk of the ectodomain is shed from the parasite surface in the form of two soluble fragments of 44 and 48 kDa. PfAMA 1 is not detectably modified by the addition of N-linked oligosaccharides. PMID- 11399765 TI - Distinct functional surface regions on ubiquitin. AB - The characterized functions of the highly conserved polypeptide ubiquitin are to target proteins for proteasome degradation or endocytosis. The formation of a polyubiquitin chain of at least four units is required for efficient proteasome binding. By contrast, monoubiquitin serves as a signal for the endocytosis of plasma membrane proteins. We have defined surface residues that are important for ubiquitin's vital functions in Saccharomyces cerevisiae. Surprisingly, alanine scanning mutagenesis showed that only 16 of ubiquitin's 63 surface residues are essential for vegetative growth in yeast. Most of the essential residues localize to two hydrophobic clusters that participate in proteasome recognition and/or endocytosis. The others reside in or near the tail region, which is important for conjugation and deubiquitination. We also demonstrate that the essential residues comprise two distinct functional surfaces: residues surrounding Phe(4) are required for endocytosis, whereas residues surrounding Ile(44) are required for both endocytosis and proteasome degradation. PMID- 11399766 TI - Functional interaction of p53 and BLM DNA helicase in apoptosis. AB - The Bloom syndrome (BS) protein, BLM, is a member of the RecQ DNA helicase family that also includes the Werner syndrome protein, WRN. Inherited mutations in these proteins are associated with cancer predisposition of these patients. We recently discovered that cells from Werner syndrome patients displayed a deficiency in p53 mediated apoptosis and WRN binds to p53. Here, we report that analogous to WRN, BLM also binds to p53 in vivo and in vitro, and the C-terminal domain of p53 is responsible for the interaction. p53-mediated apoptosis is defective in BS fibroblasts and can be rescued by expression of the normal BLM gene. Moreover, lymphoblastoid cell lines (LCLs) derived from BS donors are resistant to both gamma-radiation and doxorubicin-induced cell killing, and sensitivity can be restored by the stable expression of normal BLM. In contrast, BS cells have a normal Fas-mediated apoptosis, and in response to DNA damage normal accumulation of p53, normal induction of p53 responsive genes, and normal G(1)-S and G(2)-M cell cycle arrest. BLM localizes to nuclear foci referred to as PML nuclear bodies (NBs). Cells from Li-Fraumeni syndrome patients carrying p53 germline mutations and LCLs lacking a functional p53 have a decreased accumulation of BLM in NBs, whereas isogenic lines with functional p53 exhibit normal accumulation. Certain BLM mutants (C1055S or Delta133-237) that have a reduced ability to localize to the NBs when expressed in normal cells can impair the localization of wild type BLM to NBs and block p53-mediated apoptosis, suggesting a dominant negative effect. Taken together, our results indicate both a novel mechanism of p53 function by which p53 mediates nuclear trafficking of BLM to NBs and the cooperation of p53 and BLM to induce apoptosis. PMID- 11399767 TI - The receptor interaction region of Tyk2 contains a motif required for its nuclear localization. AB - Janus kinases have so far been viewed as enzymatic intermediates that couple a variety of cell surface receptors to downstream substrates with diverse effector functions. Tyk2 is a member of this family that is involved in the interferon alpha/beta and interleukin-12 signaling pathways via its specific interaction with the IFNAR1 and the beta1 receptor subunits. Here, we have analyzed the subcellular distribution of the wild-type Tyk2 protein and of several mutants expressed in Tyk2-deficient human cells. Direct GFP-associated fluorescence and immunostaining showed a diffuse localization of Tyk2 throughout the cell, including the nuclear compartment. The nuclear localization of Tyk2 requires a nuclear localization signal-like motif rich in arginine residues that maps within the region mediating interaction with cytokine receptors. To address the question of the role of the Tyk2 nuclear pool in interferon-alpha/beta-induced biological effects, cells expressing a membrane-targeted form of Tyk2-green fluorescent protein were analyzed for their interferon-alpha responses. Our studies demonstrate that Tyk2 can reside in the nucleus independently of receptor binding and that the nuclear pool is dispensable for the transcriptional and anti vesicular stomatitis virus responses induced by interferon-alpha. PMID- 11399768 TI - Pro-apoptotic cleavage products of Bcl-xL form cytochrome c-conducting pores in pure lipid membranes. AB - During apoptotic cell death, cells usually release apoptogenic proteins such as cytochrome c from the mitochondrial intermembrane space. If Bcl-2 family proteins induce such release by increasing outer mitochondrial membrane permeability, then the pro-apoptotic, but not anti-apoptotic activity of these proteins should correlate with their permeabilization of membranes to cytochrome c. Here, we tested this hypothesis using pro-survival full-length Bcl-x(L) and pro-death Bcl x(L) cleavage products (DeltaN61Bcl-x(L) and DeltaN76Bcl-x(L)). Unlike Bcl-x(L), DeltaN61Bcl-x(L) and DeltaN76Bcl-x(L) caused the release of cytochrome c from mitochondria in vivo and in vitro. Recombinant DeltaN61Bcl-x(L) and DeltaN76Bcl x(L), as well as Bcl-x(L), cleaved in situ by caspase 3-possessed intrinsic pore forming activity as demonstrated by their ability to efficiently permeabilize pure lipid vesicles. Furthermore, only DeltaN61Bcl-x(L) and DeltaN76Bcl-x(L), but not Bcl-x(L), formed pores large enough to release cytochrome c and to destabilize planar lipid bilayer membranes through reduction of pore line tension. Because Bcl-x(L) and its C-terminal cleavage products bound similarly to lipid membranes and formed oligomers of the same size, neither lipid affinity nor protein-protein interactions appear to be solely responsible for the increased membrane-perturbing activity elicited by Bcl-x(L) cleavage. Taken together, these data are consistent with the hypothesis that Bax-like proteins oligomerize to form lipid-containing pores in the outer mitochondrial membrane, thereby releasing intermembrane apoptogenic factors into the cytosol. PMID- 11399769 TI - The independent cue and cus systems confer copper tolerance during aerobic and anaerobic growth in Escherichia coli. AB - Copper is essential but can be toxic even at low concentrations. Coping with this duality requires multiple pathways to control intracellular copper availability. Three copper-inducible promoters, controlling expression of six copper tolerance genes, were recently identified in Escherichia coli. The cue system employs an inner membrane copper transporter, whereas the cus system includes a tripartite transporter spanning the entire cell envelope. Although cus is not essential for aerobic copper tolerance, we show here that a copper-sensitive phenotype can be observed when cus is inactivated in a cueR background. Furthermore, a clear copper-sensitive phenotype for the cus system is revealed in the absence of O(2). These results indicate that the cue pathway, which includes a copper exporter, CopA, and a periplasmic oxidase, CueO, is the primary aerobic system for copper tolerance. During anaerobic growth, however, copper toxicity increases, and the independent cus copper exporter is also necessary for full copper tolerance. We conclude that the cytosolic (CueR) and periplasmic (CusRS) sensor systems differentially regulate copper export systems in response to changes in copper and oxygen availability. These results underscore the increased toxicity of copper under anaerobic conditions and the complex adaptation of copper export in E. coli. PMID- 11399770 TI - The heterotrimer of the membrane-peripheral components of transhydrogenase and the alternating-site mechanism of proton translocation. AB - Transhydrogenase undergoes conformational changes to couple the redox reaction between NAD(H) and NADP(H) to proton translocation across a membrane. The protein comprises three components: dI, which binds NAD(H); dIII, which binds NADP(H); and dII, which spans the membrane. Experiments using isothermal titration calorimetry, analytical ultracentrifugation, and small angle x-ray scattering show that, as in the crystalline state, a mixture of recombinant dI and dIII from Rhodospirillum rubrum transhydrogenase readily forms a dI(2)dIII(1) heterotrimer in solution, but we could find no evidence for the formation of a dI(2)dIII(2) tetramer using these techniques. The asymmetry of the complex suggests that there is an alternation of conformations at the nucleotide-binding sites during proton translocation by the complete enzyme. The characteristics of nucleotide interaction with the isolated dI and dIII components and with the dI(2)dIII(1) heterotrimer were investigated. (a) The rate of release of NADP(+) from dIII was decreased 5-fold when the component was incorporated into the heterotrimer. (b) The binding affinity of one of the two nucleotide-binding sites for NADH on the dI dimer was decreased about 17-fold in the dI(2)dIII(1) complex; the other binding site was unaffected. These observations lend strong support to the alternating-site mechanism. PMID- 11399771 TI - Reflections of a fortunate biochemist. PMID- 11399772 TI - Characterization of glutathione amide reductase from Chromatium gracile. Identification of a novel thiol peroxidase (Prx/Grx) fueled by glutathione amide redox cycling. AB - Among the Chromatiaceae, the glutathione derivative gamma-l-glutamyl-l cysteinylglycine amide, or glutathione amide, was reported to be present in facultative aerobic as well as in strictly anaerobic species. The gene (garB) encoding the central enzyme in glutathione amide cycling, glutathione amide reductase (GAR), has been isolated from Chromatium gracile, and its genomic organization has been examined. The garB gene is immediately preceded by an open reading frame encoding a novel 27.5-kDa chimeric enzyme composed of one N terminal peroxiredoxin-like domain followed by a glutaredoxin-like C terminus. The 27.5-kDa enzyme was established in vitro to be a glutathione amide-dependent peroxidase, being the first example of a prokaryotic low molecular mass thiol dependent peroxidase. Amino acid sequence alignment of GAR with the functionally homologous glutathione and trypanothione reductases emphasizes the conservation of the catalytically important redox-active disulfide and of regions involved in binding the FAD prosthetic group and the substrates glutathione amide disulfide and NADH. By establishing Michaelis constants of 97 and 13.2 microm for glutathione amide disulfide and NADH, respectively (in contrast to K(m) values of 6.9 mm for glutathione disulfide and 1.98 mm for NADPH), the exclusive substrate specificities of GAR have been documented. Specificity for the amidated disulfide cofactor partly can be explained by the substitution of Arg-37, shown by x-ray crystallographic data of the human glutathione reductase to hydrogen-bond one of the glutathione glycyl carboxylates, by the negatively charged Glu-21. On the other hand, the preference for the unusual electron donor, to some extent, has to rely on the substitution of the basic residues Arg-218, His-219, and Arg-224, which have been shown to interact in the human enzyme with the NADPH 2'-phosphate group, by Leu-197, Glu-198, and Phe-203. We suggest GAR to be the newest member of the class I flavoprotein disulfide reductase family of oxidoreductases. PMID- 11399773 TI - Effect of 2'-5' phosphodiesters on DNA transesterification by vaccinia topoisomerase. AB - Vaccinia topoisomerase forms a covalent DNA-(3'-phosphotyrosyl)-enzyme intermediate at a pentapyrimidine target site 5'-CCCTTp downward arrow in duplex DNA. By introducing single 2'-5' phosphodiesters in lieu of a standard 3'-5' phosphodiester linkage, we illuminate the contributions of phosphodiester connectivity to DNA transesterification. We find that the DNA cleavage reaction was slowed by more than six orders of magnitude when a 2'-5' linkage was present at the scissile phosphodiester (CCCTT(2')p downward arrow(5')A). Thus, vaccinia topoisomerase is unable to form a DNA-(2'-phosphotyrosyl)-enzyme intermediate. We hypothesize that the altered geometry of the 2'-5' phosphodiester limits the ability of the tyrosine nucleophile to attain a requisite, presumably apical orientation with respect to the 5'-OH leaving group. A 2'-5' phosphodiester located to the 3' side of the cleavage site (CCCTTp downward arrowN(2')p(5')N) reduced the rate of transesterification by a factor of 500. In contrast, 2'-5' phosphodiesters at four other sites in the scissile strand (TpCGCCCTpT downward arrowATpTpC) and five positions in the nonscissile strand (3'-GGGpApApTpApA) had no effect on transesterification rate. The DNAs containing 2'-5' phosphodiesters were protected from digestion by exonuclease III. We found that exonuclease III was consistently arrested at positions 1 and 2 nucleotides prior to the encounter of its active site with the modified 2'-5' phosphodiester and that the 2'-5' linkage itself was poorly hydrolyzed by exonuclease III. PMID- 11399774 TI - Synergistic transcriptional activation of human Acyl-coenzyme A: cholesterol acyltransterase-1 gene by interferon-gamma and all-trans-retinoic acid THP-1 cells. AB - Acyl-coenzyme A:cholesterol acyltransferase (ACAT) is an intracellular enzyme involved in cellular cholesterol homeostasis and in atherosclerotic foam cell formation. Human ACAT-1 gene contains two promoters (P1 and P7), each located in a different chromosome (1 and 7) (Li, B. L., Li, X. L., Duan, Z. J., Lee, O., Lin, S., Ma, Z. M., Chang, C. C., Yang, X. Y., Park, J. P., Mohandas, T. K., Noll, W., Chan, L., and Chang, T. Y. (1999) J. Biol Chem. 274, 11060-11071). Interferon-gamma (IFN-gamma), a cytokine that exerts many pro-atherosclerotic effects in vivo, causes up-regulation of ACAT-1 mRNA in human blood monocyte derived macrophages and macrophage-like cells but not in other cell types. To examine the molecular nature of this observation, we identified within the ACAT-1 P1 promoter a 159-base pair core region. This region contains 4 Sp1 elements and an IFN-gamma activated sequence (GAS) that overlaps with the second Sp1 element. In the monocytic cell line THP-1 cell, the combination of IFN-gamma and all-trans retinoic acid (a known differentiation agent) enhances the ACAT-1 P1 promoter but not the P7 promoter. Additional experiments showed that all-trans-retinoic acid causes large induction of the transcription factor STAT1, while IFN-gamma causes activation of STAT1 such that it binds to the GAS/Sp1 site in the ACAT-1 P1 promoter. Our work provides a molecular mechanism to account for the effect of IFN-gamma in causing transcriptional activation of ACAT-1 in macrophage-like cells. PMID- 11399775 TI - Phosphorylation of a novel myosin binding subunit of protein phosphatase 1 reveals a conserved mechanism in the regulation of actin cytoskeleton. AB - The myotonic dystrophy kinase-related kinases RhoA binding kinase and myotonic dystrophy kinase-related Cdc42 binding kinase (MRCK) are effectors of RhoA and Cdc42, respectively, for actin reorganization. Using substrate screening in various tissues, we uncovered two major substrates, p130 and p85, for MRCKalpha kinase. p130 is identified as myosin binding subunit p130, whereas p85 is a novel related protein. p85 contains N-terminal ankyrin repeats, an alpha-helical C terminus with leucine repeats, and a centrally located conserved motif with the MRCKalpha-kinase phosphorylation site. Like MBS130, p85 is specifically associated with protein phosphatase 1delta (PP1delta), and this requires the N terminus, including the ankyrin repeats. This association is required for the regulation of both the catalytic activities and the assembly of actin cytoskeleton. The N terminus, in association with PP1delta, is essential for actin depolymerization, whereas the C terminus antagonizes this action. The C terminal effects consist of two independent events that involved both the conserved phosphorylation inhibitory motif and the alpha-helical leucine repeats. The former was able to interact with PP1delta only in the phosphorylated state and result in inactivation of PP1delta activity. This provides further evidence that phosphorylation of a myosin binding subunit protein by specific kinases confers conformational changes in a highly conserved region that plays an essential role in the regulation of its catalytic subunit activities. PMID- 11399776 TI - Caspase-2-induced apoptosis is dependent on caspase-9, but its processing during UV- or tumor necrosis factor-dependent cell death requires caspase-3. AB - Mammalian caspases are a family of cysteine proteases that plays a critical role in apoptosis. We have analyzed caspase-2 processing in human cell lines containing defined mutations in caspase-3 and caspase-9. Here we demonstrate that caspase-2 processing, during cell death induced by UV irradiation, depends both on caspase-9 and caspase-3 activity, while, during TNF-alpha-dependent apoptosis, capase-2 processing is independent of caspase-9 but still requires caspase-3. In vitro procaspase-2 is the preferred caspase cleaved by caspase-3, while caspase-7 cleaves procaspase-2 with reduced efficiency. We have also demonstrated that caspase-2-mediated apoptosis requires caspase-9 and that cells co-expressing caspase-2 and a dominant negative form of caspase-9 are impaired in activating a normal apoptotic response and release cytochrome c into the cytoplasm. Our findings suggest a role played by caspase-2 as a regulator of the mitochondrial integrity and open questions on the mechanisms responsible for its activation during cell death. PMID- 11399777 TI - Kinase insert domain receptor (KDR) extracellular immunoglobulin-like domains 4-7 contain structural features that block receptor dimerization and vascular endothelial growth factor-induced signaling. AB - The vascular endothelial growth factor (VEGF) receptor tyrosine kinase subtype kinase insert domain receptor (KDR) contains seven extracellular Ig-like domains, of which the three most amino-terminal contain the necessary structural features required for VEGF binding. To clarify the functional role of KDR Ig-like domains 4-7, we compared VEGF-induced signaling in human embryonic kidney and porcine aortic endothelial cells expressing native versus mutant receptor proteins in which Ig-like domains 4-7, 4-6, or 7 had been deleted. Western blotting using an anti-receptor antibody indicated equivalent expression levels for each of the recombinant proteins. As expected, VEGF treatment robustly augmented native receptor autophosphorylation. In contrast, receptor autophosphorylation, as well as downstream signaling events, were VEGF-independent for cells expressing mutant receptors. (125)I-VEGF(165) bound with equal or better affinity to mutant versus native receptor, although the number of radioligand binding sites was significantly reduced because a significant percentage of mutant, but not native, receptors were localized to the cell interior. As was the case for native KDR, (125)I-VEGF(165) binding to the mutant receptors was dependent upon cell surface heparan sulfate proteoglycans, and (125)I-VEGF(121) bound with an affinity equal to that of (125)I-VEGF(165) to the native and mutant receptors. It is concluded that KDR Ig-like domains 4-7 contain structural features that inhibit receptor signaling by a mechanism that is independent of neuropilin-1 and heparan sulfate proteoglycans. We speculate that this provides a cellular mechanism for blocking unwanted signaling events in the absence of VEGF. PMID- 11399778 TI - Introduction. PMID- 11399779 TI - The Trp64Arg polymorphism of the beta3-adrenergic receptor gene is not associated with training-induced changes in body composition: The HERITAGE Family Study. AB - OBJECTIVE: To investigate the association between the Trp64Arg polymorphism of the beta3-adrenergic receptor gene and changes in body composition in response to endurance training. RESEARCH METHODS AND PROCEDURES: Adult sedentary white and black subjects participating in the HERITAGE Family Study were measured before and after 20 weeks on endurance training for the body mass index, fat mass, percentage of body fat, fat-free mass, sum of eight skinfolds, and subcutaneous, visceral, and total abdominal fat areas. The association between the Trp64Arg polymorphism and the response phenotypes, computed as the difference between pre- and post-training values, was tested by analysis of covariance separately in men and women. The gene by race interaction term was also tested. RESULTS: No race differences were observed for allelic and genotype frequencies. Training resulted in significant reduction of body fat in both men and women. No association of the Trp64Arg polymorphism was observed with training-induced changes for any of the body composition phenotypes in both men and women. DISCUSSION: These results suggest that the Trp64Arg polymorphism of the beta3-adrenergic receptor gene is not related to changes in body composition in response to exercise training. PMID- 11399780 TI - Treatment of morbid obesity in inner-city women. AB - OBJECTIVE: To explore the use of the very-low-calorie formula diet (VLCD) in the indigent population of Newark, NJ, with the goal of achieving 10% weight loss within a relatively short period of 10 weeks. RESEARCH METHODS AND PROCEDURES: We accepted 131 morbidly obese indigent women into our study program. The study was limited to women only and the average starting weight was 292.3 +/- 5.9 lbs (+/- SE; 50.3 +/- 0.9 body mass index [kg/m(2)]). We used three treatment paradigms: total cost-free program for 10 weeks; cost-free, but compliance requirements; and a weekly charge of $25. The results obtained were compared with two control populations: women enrolled during the same recruitment period in a comparable suburban VLCD program and a historical control population of suburban women treated from 1985 through 1995. RESULTS: In group A (total cost-free), 79% of patients completed the 10-week program, but only 18% of patients achieved the goal of 10% weight loss. In group B when attendance and weight loss requirements were imposed, the dropout rate accelerated such that only 37% of patients completed the 10-week course, and 16% of the women were successful with their weight loss. In group C, imposing $25/wk financial outlay also accelerated dropouts but had little effect on weight loss success, which was 10% of the starting group. By comparison, the suburban patients and the historical control group exhibited 67% and 76% attendance rates after 10 weeks, and 33% and 55% success rates, respectively, in achieving the weight loss goal. DISCUSSION: We conclude that inner-city patients exhibit great interest in weight loss when financial barriers are removed. Successful weight loss was achieved in 10% to 18% of patients using the VLCD approach, approximately one-half of that obtained in affluent suburban women. Imposing financial or compliance restrictions to the inner-city patients served only to enhance dropouts. PMID- 11399781 TI - Modest lifestyle intervention and glucose tolerance in obese African Americans. AB - OBJECTIVE: Previous studies have demonstrated the benefit of short-term diets on glucose tolerance in obese individuals. The purpose of this study was to evaluate the effectiveness of modest lifestyle changes in maintaining improvements in glucose tolerance induced by short-term energy restriction in obese African Americans with impaired glucose tolerance or type 2 diabetes mellitus. RESEARCH METHODS AND PROCEDURES: An intervention group (n = 45; 47 +/- 1 year [mean +/- SE]), 105 +/- 4 kg; body mass index: 39 +/- 1 kg/m(2)) received an energy restricted diet (943 +/- 26 kcal/d) for 1 week, followed by a lifestyle program of reduced dietary fat (-125 kcal/d) and increased physical activity (+125 kcal/d) for 1 year. Body weight and plasma concentrations of glucose, insulin, and C-peptide during an oral glucose tolerance test were measured at baseline, 1 week, and 4-month intervals. A control group (n = 24; 48 +/- 1 year; 110 +/- 5 kg; body mass index: 41 +/- 2 kg/m(2)) underwent these measurements at 4-month intervals. RESULTS: No changes in weight or glucose tolerance were observed in the control group. The intervention group had significant (p < 0.05) improvements in body weight and glucose tolerance in response to the 1-week diet, which persisted for 4 months (p < 0.001 vs. control for change in weight). A total of 19 subjects (42%) continued the intervention program for 1 year, with sustained improvements (weight: -4.6 +/- 1.0 kg; p < 0.001 vs. control; oral glucose tolerance test glucose area: -103 +/- 44 mM. min; p < 0.05 vs. control). DISCUSSION: A modest lifestyle program facilitates weight loss and enables improvements in glucose tolerance to be maintained in obese individuals with abnormal glucose tolerance. However, attrition was high, despite the mild nature of the program. PMID- 11399782 TI - Weight loss attempts and attitudes toward body size, eating, and physical activity in American Indian children: relationship to weight status and gender. AB - OBJECTIVE: This study examined dieting, weight perceptions, and self-efficacy to eat healthy foods and engage in physical activity and their relationships to weight status and gender among American Indian elementary schoolchildren. RESEARCH METHODS AND PROCEDURES: Data for this study were collected as part of the baseline examination for the Pathways study. Participants were 1441 second- through third-grade American Indian children in 41 schools representing seven tribes in Arizona, New Mexico, and South Dakota who filled out a questionnaire and had heights and weights taken. RESULTS: Forty-two percent of the children were overweight or obese. No differences were found between overweight/obese and normal weight children for healthy food intentions or self-efficacy. Heavier children (especially those with body mass index > 95th percentile) were more likely to have tried to lose weight or were currently trying to lose weight. No gender differences were found. Normal weight children chose a slightly heavier body size as most healthy compared with overweight/obese children. DISCUSSION: The results indicate that children are concerned about their weight and that weight modification efforts are common among overweight American Indian children. School, community, and family-based programs are needed to help young people adopt lifelong healthful eating and physical activity practices. PMID- 11399783 TI - Comparative evaluation of fecal fat excretion induced by orlistat and chitosan. AB - OBJECTIVE: To compare the effects of chitosan and orlistat on fecal fat excretion. RESEARCH METHODS AND PROCEDURE: A randomized, open-label, two-period sequential design study was used. A total of 12 healthy adult volunteers within 20% of their ideal body weight entered a 7-day run-in diet period before being randomized to orlistat (120 mg) or chitosan (890 mg) three times daily for 7 days. Subjects then crossed over treatment regimens for an additional 7-day period. Subjects followed a standardized diet (2500 kcal/d, 30% as fat) for the entire 21-day study. Feces were collected on days 4 to 7 of the run-in period (baseline) and during the two treatment periods. Mean daily fecal fat excretion was measured at baseline and during each treatment regimen. RESULTS: Mean baseline fecal fat excretion for all subjects was 1.36 +/- 0.45 g/d. During orlistat treatment, mean fecal fat excretion significantly increased from baseline (+16.13 +/- 7.27 g/d; p < 0.001). No significant effect was observed with chitosan (+0.27 +/- 1.02 g/d; p = 0.379). Fecal fat excretion was significantly greater with orlistat than with chitosan (p < 0.001; 95% confidence intervals: 11.73; 20.00 g/d). DISCUSSION: This study provides additional evidence of the inhibitory effect of orlistat on dietary fat absorption. Chitosan, however, has no effect on fecal fat excretion. PMID- 11399784 TI - Assessment of free-living physical activity in humans: an overview of currently available and proposed new measures. AB - The number of physical activity measures and indexes used in the human literature is large and may result in some difficulty for the average investigator to choose the most appropriate measure. Accordingly, this review is intended to provide information on the utility and limitations of the various measures. Its primary focus is the objective assessment of free-living physical activity in humans based on physiological and biomechanical methods. The physical activity measures have been classified into three categories: Measures based on energy expenditure or oxygen uptake, such as activity energy expenditure, activity-related time equivalent, physical activity level, physical activity ratio, metabolic equivalent, and a new index of potential interest, daytime physical activity level. Measures based on heart rate monitoring, such as net heart rate, physical activity ratio heart rate, physical activity level heart rate, activity-related time equivalent, and daytime physical activity level heart rate. Measures based on whole-body accelerometry (counts/U time). Quantification of the velocity and duration of displacement in outdoor conditions by satellites using the Differential Global Positioning System may constitute a surrogate for physical activity, because walking is the primary activity of man in free-living conditions. A general outline of the measures and indexes described above is presented in tabular form, along with their respective definition, usual applications, advantages, and shortcomings. A practical example is given with typical values in obese and non-obese subjects. The various factors to be considered in the selection of physical activity methods include experimental goals, sample size, budget, cultural and social/environmental factors, physical burden for the subject, and statistical factors, such as accuracy and precision. It is concluded that no single current technique is able to quantify all aspects of physical activity under free-living conditions, requiring the use of complementary methods. In the future, physical activity sensors, which are of low cost, small-sized, and convenient for subjects, investigators, and clinicians, are needed to reliably monitor, during extended periods in free-living situations, small changes in movements and grade as well as duration and intensity of typical physical activities. PMID- 11399785 TI - Redox redux: a closer look at conceptal low molecular weight thiols. PMID- 11399786 TI - Carshalton in the 1960s. PMID- 11399787 TI - Diesel emissions: is more health research still needed? AB - It can legitimately be asked whether we need any more research on the health effects of diesel emissions. However, despite a research effort spanning at least 5 decades and the generation of a huge literature, there are still key uncertainties about the health impacts of present and future diesel emissions. This article briefly characterizes current knowledge and information gaps, and then proposes some key issues requiring further research. These issues include the adjuvant effect, the bioactivity of inhaled emissions at realistic doses, the toxicity of aged diesel exhaust particles, the importance of ultrafine particulate emissions, the need to improve our ability to predict the impacts of changes in emissions, and the placement of diesel health risks in context regarding other exposures. PMID- 11399788 TI - Dose-dependent nonlinear pharmacokinetics of ethylene glycol metabolites in pregnant (GD 10) and nonpregnant Sprague-Dawley rats following oral administration of ethylene glycol. AB - The kinetics of orally administered ethylene glycol (EG) and its major metabolites, glycolic acid (GA) and oxalic acid (OX), in pregnant (P; gestation day 10 at dosing, GD 10) rats were compared across doses, and between pregnant and nonpregnant (NP) rats. Groups of 4 jugular vein-cannulated female rats were administered 10 (P and NP), 150 (P), 500 (P), 1000 (P), or 2500 (P and NP) mg (13)C-labelled EG/kg body weight. Serial blood samples and urine were collected over 24-hr postdosing, and analyzed for EG, GA, and OX using GC/MS techniques. Pharmacokinetic parameters including Cmax, Tmax, AUC, and betat((1/2)) were determined for EG and GA. Pregnancy status (GD 10-11) had no impact on the pharmacokinetic parameters investigated. Blood levels of GA were roughly dose proportional from 10 to 150 mg EG/kg, but increased disproportionately from 500 to 1000 mg EG/kg. EG and GA exhibited dose-dependent urinary elimination at doses > or = 500 mg EG/kg, probably due to saturation of metabolic conversion of EG to GA, and of GA to downstream metabolites. The shift to nonlinear kinetics encompassed the NOEL (500 mg EG/kg) and LOEL (1000 mg EG/kg) for developmental toxicity of EG in rats, providing additional evidence for the role of GA in EG developmental toxicity. The peak maternal blood concentration of GA associated with the LOEL for developmental toxicity in the rat was quite high (363 microg/g or 4.8 mM blood). OX was a very minor metabolite in both blood and urine at all dose levels, suggesting that OX is not important for EG developmental toxicity. PMID- 11399789 TI - Differential activation of hepatic NF-kappaB in rats and hamsters by the peroxisome proliferators Wy-14,643, gemfibrozil, and dibutyl phthalate. AB - Nuclear factor-kappaB (NF-kappaB) is an oxidative stress-activated transcription factor involved in the regulation of cell proliferation and apoptosis. We found previously that the peroxisome proliferator ciprofibrate activates NF-kappaB in the livers of rats and mice. These species are sensitive to the hepatocarcinogenic effects of peroxisome proliferators, whereas other species such as Syrian hamsters are not. In the present study we examined the effects of 3 different peroxisome proliferators on NF-kappaB activation in rats and Syrian hamsters. The peroxisome proliferators Wy-14,643, gemfibrozil, and dibutyl phthalate were administered to animals for 6, 34, or 90 days. NF-kappaB activity was determined using electrophoretic mobility-shift assays and confirmed using supershift assays. Wy-14,643 increased the DNA binding activity of NF-kappaB at all 3 time points in rats and produced the highest activation of the 3 chemicals tested. Gemfibrozil and dibutyl phthalate increased NF-kappaB activation to a lesser extent in rats and not at all times. There were no differences in hepatic NF-kappaB levels between control hamsters and hamsters treated with any of the peroxisome proliferators. This study demonstrates species-specific differences in hepatic NF-kappaB activation by peroxisome proliferators. PMID- 11399790 TI - Wy-14,643-induced hypomethylation of the c-myc gene in mouse liver. AB - The carcinogenic activity of Wy-14,643 in mouse liver appears to be nongenotoxic and could involve a decrease in DNA methylation. The mechanism for Wy-14,643 induced decrease in DNA methylation is proposed to involve increased cell proliferation followed by prevention of the methylation of the newly synthesized DNA. To investigate this mechanism, female B6C3F1 mice were administered daily by oral gavage 50 mg/kg Wy-14,643. Mice were sacrificed at 2, 5, 8, 24, 26, 29, 32, 36, 48, 72, and 96 h after the first dose. Some mice also received 450 mg/kg methionine by ip injection at 30 min after administering Wy-14,643. Hypomethylation of the c-myc gene first occurred at 48 h after the first dose of Wy-14,643. Cell proliferation determined by the Proliferating Cell Nuclear Antigen (PCNA)-Labeling Index started to increase at 36 h and peaked at 72h. Wy14,643 did not affect the liver concentration of either S-adenosyl methionine (SAM) or S-adenosyl homocysteine (SAH). Methionine prevented and reversed the hypomethylation of the c-myc gene induced by Wy-14,643. However, the increased levels of SAM and SAH returned to control levels prior to the prevention by methionine of Wy-14,643-induced hypomethylation. Furthermore, methionine did not prevent Wy-14,643-induced increase in the PCNA-Labeling Index. The activity of nuclear DNA methyltransferase (DNA MTase) was increased at 72 and 96 h after administering Wy14,643. Wy14,643 also increased the activity of DNA MTase when added in vitro to nuclear extracts. The results are consistent with Wy-14,643 decreasing the methylation of the c-myc gene by a mechanism that includes enhancement of cell proliferation followed by prevention of the methylation of the newly synthesized DNA. However, the results indicate that Wy-14,643 does not prevent methylation by decreasing either the availability of SAM or the activity of DNA MTase. PMID- 11399791 TI - Hepatocellular iron accumulation and increased cell proliferation in polychlorinated biphenyl-exposed Sprague-Dawley rats and the development of hepatocarcinogenesis. AB - Polychlorinated biphenyls (PCBs) are liver-tumor promoters in rodents, but the underlying mechanisms have not been fully elucidated. Tissue sections from the PCB bioassay reported by Mayes et al. 1998, Toxicol Sci., 41-66, were evaluated by histopathological techniques that included immunohistochemistry. In females, and to a much lesser extent in males, iron accumulation in hepatocytes was found at the 26th-week sacrifice, which was pronounced in the mid- and high-dose Aroclor-1254 and -1260 groups. At 52 weeks, large accumulations of iron were also present in Kupffer cells of females, and dose-related increases in proliferating cell nuclear antigen (PCNA) hepatocyte labeling indices were found in both males and females. These changes preceded the formation of liver tumors, which were not generally found until 78 weeks. Glutathione S-transferase placental (GSTP) positive foci were present at 52 weeks in high-dose Aroclor-1254 and -1260 female groups, and small foci were found in some Aroclor 1254-exposed female rats at 26 weeks, along with centrilobular hepatocytes expressing GSTP. The results of this study suggest that PCB-induced iron accumulation in hepatocytes is an early event that may be related to tumor formation, especially in female rats. In both males and females, increases in cell proliferation at 52 weeks were statistically significantly correlated with tumor incidences at termination among the various PCB dosage groups. Consequently, iron accumulations producing oxidative damage, and enhanced cell proliferation resulting in tumor promotion may be components in the mode of action for PCB-induced hepatocarcinogenesis in rodents. PMID- 11399792 TI - In utero and lactational treatment with 2,3,7,8-tetrachlorodibenzo-p-dioxin impairs mammary gland differentiation but does not block the response to exogenous estrogen in the postpubertal female rat. AB - These experiments tested whether in utero and lactational exposure to 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) alters mammary gland differentiation, estrogen receptor alpha (ERalpha) expression levels, or the response to estrogen in the female postpubertal rat mammary gland. Pregnant Holtzman rats were administered a single oral dose of 1 microg/kg TCDD or vehicle on gestation-day 15. Exposed and non-exposed female offspring were weaned on postnatal day 21 and ovariectomized at 9 weeks of age. Two weeks later, both TCDD and control animals were divided into 3 groups, receiving treatment with placebo, 0.025, or 0.1 mg 17beta estradiol pellet implants. After 48 h, mammary tissue was removed for analysis following euthanasia. TCDD-exposed mammary glands demonstrated impaired differentiation as measured by the distribution of terminal ductal structures and increased expression levels of ERalpha. The response to exogenous estrogen was tested in TCDD-exposed animals and compared to control non-exposed animals. Estrogen stimulation of the TCDD-exposed glands induced progesterone receptor expression and mammary gland differentiation as measured by a shift in distribution from terminal end buds and terminal ducts to Types I and II lobules. Control glands were better differentiated at baseline and did not exhibit any significant changes in the distribution of terminal ductal structures following estrogen stimulation. The increase in progesterone receptor-expression levels by exogenous estrogen in control glands was similar to the TCDD-exposed glands. These experiments demonstrate that in utero and lactational exposures to TCDD impair mammary gland differentiation but that TCDD-exposed mammary glands retain the ability to differentiate in response to estrogen. PMID- 11399793 TI - Induction of glutathione S-transferase activity and protein expression in brown bullhead (Ameiurus nebulosus) liver by ethoxyquin. AB - The inducibility of hepatic cytosolic glutathione S-transferases (GSTs) was examined in brown bullheads, a freshwater fish that is highly susceptible to hepatic neoplasia following exposure to carcinogen-contaminated sediments. Juvenile bullheads were fed a semi-purified antioxidant-free diet supplemented with ethoxyquin (0.5% w/w dissolved in 3% corn oil), a prototypical rodent GST inducing agent, twice daily for 14 days. Control bullheads received the antioxidant-free diet supplemented with corn oil (3% w/w). A significant increase (1.6-fold, p < or = 0.01) in hepatic cytosolic GST activity toward 1-chloro-2,4 dinitrobenzene (CDNB) was observed in the ethoxyquin-treated bullheads relative to control fish. A trend toward increased GST-NBC activity was observed in the ethoxyquin-treated fish (1.2-fold, p = 0.06), whereas no treatment-related effects were observed on GST activities toward ethacrynic acid (ECA). In contrast, GST activity toward (+/-)-anti-benzo[a]pyrene-trans-7,8-dihydrodiol 9,10-epoxide (BPDE) was repressed in affinity-purified cytosolic fractions prepared from ethoxyquin-treated bullheads relative to control bullheads. Silver staining and densitometric analysis of isoelectric-focused, affinity-purified GST proteins revealed increased expression of two basic GST-like isoforms in ethoxyquin-treated fish. In summary, exposure to ethoxyquin increases brown bullhead GST-CDNB catalytic activity and hepatic cationic GST protein expression. However, the increase in overall GST-CDNB activity by ethoxyquin is associated with repression of GST-BPDE activity, suggesting differential effects on hepatic bullhead GST isoforms by ethoxyquin. The potential repression of bullhead GST isoforms that conjugate the carcinogenic metabolites of PAH metabolism under conditions of environmental chemical exposure could be a contributing factor in the sensitivity of bullheads to pollutant-associated neoplasia. PMID- 11399794 TI - In vivo manipulation of endogenous metallothionein with a monoclonal antibody enhances a T-dependent humoral immune response. AB - Metallothionein (MT) is a small stress response protein that can be induced by exposure to heavy metal cations, oxidative stressors, and acute phase cytokines that mediate inflammation. In previous experiments, we have shown that exogenous MT can affect cell proliferation, macrophage and cytotoxic T lymphocyte function, and humoral immunity to T-dependent antigens. In the studies described here, we have explored the effect of a monoclonal anti-MT antibody (clone UC1MT) on the role that endogenous MT plays in the humoral immune response. In vivo injection of UC1MT significantly increased the humoral response to simultaneous challenge with ovalbumin (OVA). In contrast, mice immunized with OVA in the presence of an isotype-matched antibody control (MOPC 21) showed no change in the anti-OVA humoral response. The predominant anti-OVA response that was enhanced by UC1MT treatment was the IgG(1) response; the IgG(2a) anti-OVA response was not altered by UC1MT treatment. UC1MT treatment increased the numbers of IgG anti-OVA secreting cells as measured by ELISPOT assay, suggesting that blocking the effects of MT synthesized during the immune response augments the differentiation of antigen-specific plasma cells. The percentages of T and B cells in the spleens of animals from each treatment group were not significantly different, suggesting that this regimen of UC1MT treatment does not significantly affect hematopoiesis, but rather alters antigen-induced differentiation of lymphocytes. These observations are compatible with previous results from our laboratory that suggest that endogenous MT synthesized during the normal immune response or as a consequence of toxicant exposure suppresses in vivo immune function. In light of the fact that significant amounts of MT can be synthesized during toxicant exposure, manipulation of MT levels with an anti-MT antibody may ultimately represent an important therapeutic approach to the treatment of immune dysfunctions that result from toxicant exposure. PMID- 11399795 TI - Suppression of allergic immune responses to house dust mite (HDM) in rats exposed to 2,3,7,8-TCDD. AB - Exposure to various xenobiotics, including oxidant gases, diesel exhaust, and certain pesticides, has been reported to exacerbate pulmonary allergic hypersensitivity responses. Increased lymphocyte proliferative responses to parasite antigens or increased antibody responses to sheep erythrocyte have also been reported in rats exposed to TCDD before infection or immunization. As a result, these studies were conducted to test the hypothesis that TCDD exposure exacerbates the allergic response to house dust mite antigen. Brown Norway rats were injected, ip, with 0, 1, 10, or 30 microg TCDD/kg 7 days before intratracheal (it) sensitization to semipurified house dust mite allergen (HDM). Fourteen days later, rats were challenged with HDM and immediate bronchospasm was measured. At this time point, plus 2 and 7 days later, inflammatory cells in bronchoalveolar lavage fluid (BALF), HDM-specific IgE levels in serum, and HDM driven cell proliferation in bronchial lymph nodes and spleen were evaluated. TCDD exposure decreased both immediate bronchoconstriction and specific IgE synthesis after the HDM challenge; 7 days later, HDM-specific IgE responses remained suppressed. Total serum IgE levels were similar in all groups. HDM challenge alone significantly increased cellular and biochemical indicators of lung injury, both of which were suppressed by TCDD exposure. The proliferative response of lymph node cells, but not of spleen cells, to HDM was also suppressed at the highest TCDD dose, although the splenic response to Concanavalin A was elevated. It appears that early events in the response to HDM are affected by TCDD exposure, since message for IL5 was dramatically reduced 2 days after sensitization, but not after challenge. We therefore conclude that TCDD exposure suppressed, rather than enhanced the development of allergic immune responses and the expression of immune-mediated lung disease. PMID- 11399796 TI - Growth and development in rats given recombinant human epidermal growth factor(1 48) as neonates. AB - To assess effects of supraphysiologic doses of human recombinant epidermal growth factor(1-48) (rhEGF(1-48)) on neonatal rats, 10 litters of Wistar rats/treatment group were given 0 (formulated vehicle), 10, 100, or 1000 microg/kg daily by subcutaneous injection on postnatal days (PND) 1 through 6. Clinical signs, body weight, acquisition of developmental landmarks and reflexes, and behavior were monitored during treatment and for 5 weeks thereafter (to PND 42). A subset of animals was euthanized weekly from PND 7-28 and necropsied. Selected tissues were examined microscopically. Body weight gain at 1000 microg/kg during treatment was significantly less than control. Precocious incisor eruption, eye opening, vaginal opening, and preputial separation occurred at 100 and/or 1000 microg/kg. Acquisition of reflexes (negative geotaxis, wire maneuver, acoustic startle reflex, and visual placing) was delayed at 1000 microg/kg. Acquisition of adult locomotion was also delayed at 1000 microg/kg. These effects were transient, as locomotor activity at PND 28 and 42 did not differ from control. Effects on acoustic-startle responding persisted in females to final assessment on PND 42. Habituation to repeated acoustic stimuli was impaired, as well as response inhibition following a prepulse acoustic stimulus. rhEGF(1-48) induced structural changes in the skin, retina, kidney, oral and nasal mucosa, lung, and liver. Many of these changes were consistent with the expected mitogenic activity of rhEGF(1 48) and were transient in nature, as severity and incidence diminished with time. An exception was changes observed in the retina at 1000 microg/kg (rosettes/folds and focal defects in the outer nuclear/photoreceptor layers) that were still present 3 weeks after termination of treatment. Acceleration of developmental landmarks; suppression of reflexes, behavior, and somatic growth; and mitogenic responses in epidermal tissues have been reported in rodents treated with epidermal growth factor (EGF) derived from various mammalian species. These results demonstrate that a 48-amino acid fragment of human EGF produced by recombinant technology also induces such effects. PMID- 11399797 TI - Spatial glutathione and cysteine distribution and chemical modulation in the early organogenesis-stage rat conceptus in utero. AB - Glutathione (GSH), cysteine, and other low-molecular-weight thiols (LMWT) play a vital role in the detoxication of xenobiotics and endogenous chemicals. Differential alterations of LMWT status in various cell types of the developing embryo may underlie cell-specific sensitivity or resistance to xenobiotics and contribute to embryotoxicity. This study describes the spatial and temporal distribution of LMWTs in rat conceptuses and alterations produced by the non teratogenic GSH modulator, acetaminophen (APAP). Pregnant female rats were given 125, 250, or 500 mg/kg APAP (po) on gestational day 9. Conceptal LMWT was localized histochemically using mercury orange in cryosections, and GSH and cysteine concentrations were measured by HPLC analysis. Mercury orange histofluorescence revealed a non-uniform distribution of LMWT in untreated conceptal tissues, with strongest staining observed in the ectoplacental cone (EPC), visceral yolk sac (VYS), and embryonic heart. Less intense staining was observed in the neuroepithelium. Following treatment with APAP, tissue-associated LMWT decreased dramatically except in the EPC, while exocoelomic fluid LMWT, and LMWT within embryonic lumens, increased. Exposure to 250 mg/kg APAP decreased embryonic GSH after 6 and 24 h by 46% and 38%, respectively. Acetaminophen (500 mg/kg) decreased embryonic and VYS cysteine content by 54% and 83%, respectively, after 24 h. Acetaminophen alters the spatial distribution of LMWT in rat conceptuses, particularly with respect to cysteine. The mobilization of cysteine following chemical insult may influence the ability of conceptal cells to maintain normal GSH status due to reduced availability of cysteine for de novo GSH synthesis. PMID- 11399798 TI - Teratogenicity of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in mice lacking the expression of EGF and/or TGF-alpha. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) exposure produces hydronephrosis and cleft palate in mice. These responses are correlated with disruption of expression of epidermal growth factor (EGF) receptor ligands, primarily EGF and transforming growth factor-alpha (TGF-alpha), and altered epithelial cell proliferation and differentiation. This research examined the role of these growth factors in TCDD-induced teratogenicity by using wild type (WT) and knockout (-/-) mice that do not express EGF, TGF-alpha, or both EGF and TGF alpha. Pregnant females were weighed on GD 12 and dosed by gavage with either corn oil or TCDD at 24 microg/kg, 5 ml/kg. On GD 17.5, the maternal parameters evaluated included body weight, body weight gain, liver weight (absolute and adjusted for body weight). The number of implantations, live and dead fetuses, early or late resorptions, the proportion of males, fetal body weight, fetal absolute and relative liver weight, placenta weight, incidence of cleft palate, and the severity and incidence of hydronephrosis were recorded. TCDD did not affect maternal weight gain, fetal weight, or survival, but maternal and fetal liver weights and liver-to-body weight ratios were increased in all genotypes. The WT and TGF-alpha (-/-), but not the EGF (-/-) and EGF + TGF-alpha (-/-) fetuses, developed cleft palate after exposure to 24 microg TCDD/kg. Hydronephrosis was induced by TCDD in all genotypes, with the incidence in EGF + TGF-alpha (-/-) fetuses comparable to that of the WT. The incidence and severity of this defect was substantially increased in EGF (-/-) and TGF-alpha (-/-). In conclusion, this study demonstrated that expression of EGF influences the induction of cleft palate by TCDD. Also, EGF and TGF-alpha are not required for the induction of hydronephrosis, but when either is absent the response of the fetal urinary tract to TCDD is enhanced. PMID- 11399799 TI - alpha-Chlorohydrin inhibits glyceraldehyde-3-phosphate dehydrogenase in multiple organs as well as in sperm. AB - Numerous studies have documented inhibitory effects of alpha-chlorohydrin (ACH) on glyceraldehyde-3-phosphate dehydrogenase (G3PDH) activity in spermatozoa. A sperm-specific G3PDH isoform has been described. The possibility that ACH may inhibit G3PDH in cell types other than sperm was investigated in this work. In addition, the onset of ACH-induced epididymal toxicity was described. Changes to epididymal histology occurred 6 h following a single dose of ACH (50 mg/kg po) and were confined to the proximal initial segment. By 24 h, no epithelial cells lined the basement membrane of that region. Three h after ACH administration (50 mg/kg po), G3PDH activity was significantly decreased in sperm (85%) as well as in kidney (31%), liver (49%), and epididymis (35%). Enzyme activity remained inhibited at 6 and 24 h. G3PDH was immunolocalized in the epididymis and staining was highest in the efferent ducts and initial segment as well as in smooth muscle. Since G3PDH is a microtubule-associated protein and microtubule-dependent endocytosis occurs in the epididymis, beta-tubulin was also immunolocalized. beta tubulin densely stained the apical region of initial segment and caput epithelial cells. Disruption of beta-tubulin immunostaining correlated with the localization and onset of the lesion. Co-localization of G3PDH and beta-tubulin immunostaining was not observed although both antibodies most densely stained the initial segment. Our data indicate that histologic changes to the proximal initial segment of the epididymis occur rapidly, but subsequent to G3PDH inhibition. Moreover, ACH inhibition of G3PDH is not confined to sperm, although the sperm enzyme is most sensitive to inhibition. PMID- 11399800 TI - Proposed occupational exposure limits for select ethylene glycol ethers using PBPK models and Monte Carlo simulations. AB - Methoxyethanol (ethylene glycol monomethyl ether, EGME), ethoxyethanol (ethylene glycol monoethyl ether, EGEE), and ethoxyethyl acetate (ethylene glycol monoethyl ether acetate, EGEEA) are all developmental toxicants in laboratory animals. Due to the imprecise nature of the exposure data in epidemiology studies of these chemicals, we relied on human and animal pharmacokinetic data, as well as animal toxicity data, to derive 3 occupational exposure limits (OELs). Physiologically based pharmacokinetic (PBPK) models for EGME, EGEE, and EGEEA in pregnant rats and humans have been developed (M. L. Gargas et al., 2000, Toxicol. Appl. Pharmacol. 165, 53-62; M. L. Gargas et al., 2000, Toxicol. Appl. Pharmacol. 165, 63-73). These models were used to calculate estimated human-equivalent no adverse effect levels (NAELs), based upon internal concentrations in rats exposed to no observed effect levels (NOELs) for developmental toxicity. Estimated NAEL values of 25 ppm for EGEEA and EGEE and 12 ppm for EGME were derived using average values for physiological, thermodynamic, and metabolic parameters in the PBPK model. The uncertainties in the point estimates for the NOELs and NAELs were estimated from the distribution of internal dose estimates obtained by varying key parameter values over expected ranges and probability distributions. Key parameters were identified through sensitivity analysis. Distributions of the values of these parameters were sampled using Monte Carlo techniques and appropriate dose metrics calculated for 1600 parameter sets. The 95th percentile values were used to calculate interindividual pharmacokinetic uncertainty factors (UFs) to account for variability among humans (UF(h,pk)). These values of 1.8 for EGEEA/EGEE and 1.7 for EGME are less than the default value of 3 for this area of uncertainty. The estimated human equivalent NAELs were divided by UF(h,pk) and the default UFs for pharmacodynamic variability among animals and among humans to calculate the proposed OELs. This methodology indicates that OELs (8-h time weighted average) that should protect workers from the most sensitive adverse effects of these chemicals are 2 ppm EGEEA and EGEE (11 mg/m(3) EGEEA, 7 mg/m(3) EGEE) and 0.9 ppm (3 mg/m(3)) EGME. These recommendations assume that dermal exposure will be minimal or nonexistent. PMID- 11399801 TI - Polycystic kidney disease induced in F(1) Sprague-Dawley rats fed para nonylphenol in a soy-free, casein-containing diet. AB - para-Nonylphenol (NP; CAS #84852-15-3), an alkylphenol with a 9-carbon olefin side chain, is widely used in the manufacture of nonionic surfactants, lubricant additives, polymer stabilizers, and antioxidants. Due to its wide commercial use and putative endocrine activity in humans and wildlife, the NTP elected to assess its effects on reproduction in multigenerational studies. To avoid known estrogenic activity of phytoestrogens in soy and alfalfa, a soy- and alfalfa free, casein-containing diet was used in a range-finding study to determine the doses of NP to be tested further. NP was administered to Sprague-Dawley rats in the diet at 0, 5, 25, 200, 500, 1000, or 2000 ppm to F(0) dams beginning on gestation-day 7. The F(1) pups were weaned at postnatal day (PND) 21, and their exposure via diet was continued at the same dose level as their respective dams. Pup weights from birth through weaning were not significantly different from controls in any dose group, but the average weight of both sexes was significantly less compared to controls, beginning with the PND 28 weighing. The F(1) rats were sacrificed on PND 50 (n = 15, 3 pups of each sex from 5 litters for all dose groups). Terminal body weights of males and females in the 2000-ppm dose group were 74% and 85% of controls, respectively. Severe polycystic kidney disease (PKD) was present in 100% of the 2000 ppm-exposed male and female rats. At 1000 ppm, 67% of males and 53% of females had mild to moderate PKD versus none of either sex in the control and lower-dose groups. The no-adverse-effect level (NOAEL) for PKD was determined to be 500 ppm. Previous studies with comparable duration and route of exposure, but using soy-containing diets, reported either no or only mild PKD at 2000 ppm NP. We conclude that the renal toxicity of NP is highly dependent on the diet on which the animals are maintained. The potential interaction of diet and test compounds on nonreproductive as well as reproductive endpoints should be considered when contemplating the use of special diets formulated to minimize exogenous "hormone" content for the study of the effects of putative endocrine disruptive chemicals. PMID- 11399802 TI - Lack of effect of single high doses of buprenorphine on arterial blood gases in the rat. AB - High dose buprenorphine, a potent semisynthetic agonist-antagonist for opiate receptors, is now used in substitution treatment of human heroin addiction. Deaths have been reported in addicts misusing buprenorphine. We determined the median lethal dose (LD(50)) and studied the effects of high doses of intravenous buprenorphine on arterial blood gases in rats. Male Sprague-Dawley rats were administered buprenorphine intravenously to determine the LD(50) using the up-and down method. Subsequently, catheterized groups of 10 restrained rats received no drug, saline, acid-alcohol aqueous solvent (required to dissolve buprenorphine at a high concentration), or 3, 30, or 90 mg/kg of buprenorphine intravenously. Serial arterial blood gases were obtained over 3 h. The LD(50) determined in triplicate was 146.5 mg/kg (median of 3 series, range: 142.6-176.5). The mean dose received by surviving animals was 96.9 +/- 46.7 mg/kg. There was a significant effect of the acid-alcohol aqueous solvent on arterial blood gases. Excluding the solvent effect, 3, 30, and 90-mg/kg buprenorphine doses had no significant effects on arterial blood gases. The toxicity of intravenous buprenorphine in adult rats, assessed by the LD(50), is low. These data are consistent with a wide margin of safety of buprenorphine. The mechanism of death after the intravenous administration of a lethal dose of buprenorphine remains to be determined. PMID- 11399803 TI - Subchronic inhalation toxicity of the chlorinated propane 1,1,1,3,3,3 hexachloropropane (HCC-230fa). AB - Male and female rats were exposed by inhalation (whole body) to HCC-230fa (1,1,1,3,3,3-hexachloropropane) for 6 h/day, 5 days/week over a 15-week period. Concentrations of 0, 0.50, 2.5, and 25 ppm were studied. A total of eight groups/sex were exposed. Four groups of male and four groups of female rats were used to measure clinical signs and growth, clinical pathology, and tissue pathology. The remaining four groups of male rats were used for immunotoxicological and sperm assessment evaluations, and the remaining four groups of female rats were used for immunotoxicological evaluation. Following the exposure period, surviving male rats were kept for a 1- or 3-month recovery period. Male and female rats exposed to 25 ppm had lower mean body weights, mean body weight gains, and food consumption during the exposure period. Male and female rats exposed to 25 ppm and sacrificed immediately after the exposure period had minimally decreased total leukocyte and lymphocyte counts. These changes were considered to be marginally adverse. Pathologic examination revealed hepatocellular hypertrophy in 0-day recovery males and an increased incidence and/or severity in chronic progressive nephropathy in 0-day, 1-month recovery, and 3-month recovery males at 25 ppm. No other pathological changes, including the testis, epididymis, and other accessory sex organs, were noted in rats during the study. Evaluation of sperm parameters at the end of the exposure period showed statistically significant decreases in epididymal sperm number per cauda epididymis, percent motile sperm, and percent normal sperm morphology at 25 ppm. The biological significance of the slight changes observed in the sperm parameters in the absence of histopathological changes is unclear. After a 1 month recovery period, no biologically significant differences in sperm parameters were noted at 25 ppm compared with controls. Exposure to HCC-230fa did not significantly alter the primary humoral immune response to sheep red blood cell (SRBC). Under the conditions of this study, the no-observed-adverse-effect level (NOAEL) was considered to be 2.5 ppm. PMID- 11399804 TI - Hepatic vitamin a depletion is a sensitive marker of 2,3,7,8-tetrachlorodibenzo-p dioxin (TCDD) exposure in four rodent species. AB - 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD)-treated animals show altered retinoid homeostasis and exhibit signs of toxicity similar to those of vitamin A-deficient animals. In this study we established dose-response curves for sublethal oral doses of TCDD and hepatic vitamin A gain in four rodent species. This was done to evaluate any potential correlation between decreased hepatic vitamin A gain and other TCDD-induced effects, particularly depressed body weight gain and hepatic CYP1A induction. Young Hartley guinea pigs, Sprague-Dawley rats, C57BL/6 mice, and Golden Syrian hamsters were given single oral doses of TCDD at up to 2.5, 100, 1000, and 1000 microg/kg bw, respectively, and killed 28 days after treatment. Hepatic vitamin A gain was decreased 25% compared to controls at estimated doses of 0.1, 0.9, 1.1 and 3.6 microg/kg bw in guinea pigs, hamsters, rats, and mice, respectively. CYP1A induction and hepatic vitamin A gain were affected at similar dose levels and showed similar, but inverse dose-response curves in each of the four species, consistent with the hypothesis that altered vitamin A homeostasis is Ah-receptor mediated. In addition, there was an apparent correlation between the dose-response curves for decreased hepatic vitamin A gain and decreased body weight gain in all species. Taken together with the known importance of vitamin A in body weight regulation, this result was consistent with a contributing role for altered retinoid homeostasis in the wasting syndrome induced by TCDD. PMID- 11399805 TI - Association of quinone-induced platelet anti-aggregation with cytotoxicity. AB - Various anti-platelet drugs, including quinones, are being investigated as potential treatments for cardiovascular disease because of their ability to prevent excessive platelet aggregation. In the present investigation 3 naphthoquinones (2,3-dimethoxy-1,4-naphthoquinone [DMNQ], menadione, and 1,4 naphthoquinone [4-NQ]) were compared for their abilities to inhibit platelet aggregation, deplete glutathione (GSH) and protein thiols, and cause cytotoxicity. Platelet-rich plasma, isolated from Sprague-Dawley rats, was used for all experiments. The relative potency of the 3 quinones to inhibit platelet aggregation, deplete intracellular GSH and protein thiols, and cause cytotoxicity was 1,4-NQ > menadione >> DMNQ. Experiments using 2 thiol-modifying agents, dithiothreitol (DTT) and 1-chloro-2,4-dintrobenzene (CDNB), confirmed the key roles for GSH in quinone-induced platelet anti-aggregation and for protein thiols in quinone-induced cytotoxicity. Furthermore, the anti-aggregative effects of a group of 12 additional quinone derivatives were positively correlated with their ability to cause platelet cytotoxicity. Quinones that had a weak anti-aggregative effect did not induce cytotoxicity (measured as LDH leakage), whereas quinones that had a potent anti-aggregative effect resulted in significant LDH leakage (84 96%). In one instance, however, p-chloranil demonstrated a potent anti aggregative effect, but did not induce significant LDH leakage. This can be explained by the inability of p-chloranil to deplete protein thiols, even though intracellular GSH levels decreased rapidly. These results suggest that quinones that deplete GSH in platelets demonstrate a marked anti-aggregative effect. If this anti-aggregative effect is subsequently followed by depletion of protein thiols, cytotoxicity results. PMID- 11399807 TI - Re: androgen receptor antagonism by the organophosphate insecticide fenitrothion. PMID- 11399808 TI - Transfusion medicine illustrated. The human blood platelet, resting and active. PMID- 11399809 TI - Alloimmune platelet refractoriness: incidence declines, unsolved problems persist. PMID- 11399810 TI - The use of platelet concentrates versus plateletpheresis-the donor perspective. PMID- 11399811 TI - Incidence rates of viral infections among repeat donors:are frequent donors safer? AB - BACKGROUND: Incidence rates (IRs) for viral infections may vary with the frequency of donation among repeat, community, whole-blood (WB) donors, with IRs thought to be lower among donors with higher frequency of donation. STUDY DESIGN AND METHODS: IRs for HIV, HTLV, HCV, and HBV infection were stratified by frequency of donation among 868,403 repeat WB donors who gave approximately 4 million donations at five United States blood centers from 1991 through 96. All donors had given at least 2 donations during those years, with the first donation being nonreactive on confirmatory testing. Frequency of donation was measured in three ways: by the number of donations per year; at the time of donation, by the number of donations given within the preceding 2-year period; and by the number of donations given from 1991 through 1993. RESULTS: The IRs for HIV, HCV, and HBV infection did not appear to differ among donors with lower or higher numbers of donation per year. However, the IR for HTLV infection decreased as the number of donations per year increased (p = 0.0004). IRs for all viral markers remained stable, regardless of the number of donations given within the 2-year period before the donation. Although IRs for HIV, HTLV, and HCV infection did not vary by the number of donations given in 1991 through 1993, the IR for HBV infection appeared to be lower in donors who gave more donations in that period (p = 0.01). CONCLUSION: These findings do not provide evidence of a lower IR for transfusion transmissible viral infections among repeat WB donors who give more frequently. Abbreviated screening histories for frequent repeat donors might not be advisable. PMID- 11399812 TI - Repeat whole-blood and plateletpheresis donors:unreported deferrable risks, reactive screening tests, andresponse to incentive programs. AB - BACKGROUND: Evaluating plateletpheresis (PPH) and repeat community whole-blood (RWB) donors' responses to donation incentive programs is essential for developing effective donor retention programs. STUDY DESIGN AND METHODS: Using data from a 1998 anonymous questionnaire sent to 92,581 US blood donors, the prevalence of unreported deferrable risks, screening test reactivity, and response to incentives were compared in RWB and PPH donors by the use of weighted chi-square tests and logistic regression analyses. RESULTS: From 52,650 respondents, 38,884 RWB and 2,028 PPH donors were identified. Levels of screening test reactivity (1%) and unreported deferrable risks (UDRs, 2-3%) were similar in RWB and PPH donors. RWB and PPH donors were strongly encouraged or discouraged by similar incentives. Of the incentives that would encourage a higher proportion of UDR-free RWB donors to return, cholesterol screening and earning a blood credit appealed to >50 percent. Similar results were obtained for cholesterol screening in PPH donors. Community service or education credits, premarital screening, and cash had limited appeal for PPH and RWB donors, respectively, and would be more likely to differentially encourage donors with a UDR to return. CONCLUSION: Incentives that were associated with the greatest donor appeal and that minimized the potential recruitment of more risky donors were identified. PMID- 11399813 TI - Absence of risk factors for false-positive test results in blood donors with a reactive test result in an automated treponemal test (PK-TP) for syphilis. AB - BACKGROUND: Screening and confirmatory serologic tests for syphilis are known to generate false-positive results in low-risk populations, which include blood donors. This study assessed whether conditions previously reported to cause biological false-positive (BFP) test results for syphilis are relevant to contemporary syphilis testing of blood donors and the extent to which seropositive donors report a history of syphilis. STUDY DESIGN AND METHODS: A history of conditions reported to be associated with BFP syphilis tests or a history of syphilis infection was assessed by a case-control study of donors with reactive and nonreactive automated treponemal test results, using an anonymous mail survey. Analysis of cases was stratified by fluorescent treponemal antibody absorption (FTA-ABS) result. RESULTS: Adjusted ORs (95% CIs) for reported BFP associated conditions were 1.3 (0.8-2.1) for FTA-ABS-positive cases and 0.8 (0.3 1.9) for FTA-ABS-negative cases. Among responding blood donors, syphilis history was reported in 78 (51%) of 153 FTA-ABS-positive cases, 0 of 142 FTA-ABS-negative cases, and 3 (0.4%) of 716 automated treponemal test (PK-TP)-negative controls. CONCLUSION: Approximately half of donors with FTA-ABS-positive test results reported a syphilis history. There was no difference between reported BFP conditions for FTA-ABS-positive or FTA-ABS-negative cases and controls. This information may be useful when providing donors with better predonation or post test counseling information about syphilis testing. PMID- 11399814 TI - Seroprevalence of known and putative hepatitis markers in United States blood donors with ALT levels at least 120 IU per L. AB - BACKGROUND: ALT testing of blood donors was initiated as a surrogate marker for non-A, non-B hepatitis. Increased sensitivity of subsequent HBV and HCV tests used for standard donor screening make any residual value of ALT testing questionable. STUDY DESIGN AND METHODS: A prospective study was conducted in 166 of 645 eligible blood donors from three American Red Cross regions whose ALT was > or =120 IU per L and whose standard donor screening tests were negative. Of these enrolled donors, 124 (75%) completed follow-up. Samples obtained from the index donation, at enrollment (1 month), and at follow-up (6 months) underwent the standard donor screening tests, as well as those for HCV RNA and HGV RNA (RT PCR), antibodies to the virus envelope E2 protein of GB virus type C (GBV-C E2 antibody), and IgM antibody for CMV, parvovirus B19, EBV VCA, and HAV. Participants completed a brief demographic and exposure history questionnaire at follow-up. RESULTS: All study samples were negative in standard donor-screening tests. ALT levels were variable at return visits, with 80 to 86 percent <120 IU per L. No participants were positive for HCV RNA; 4 percent were positive for HGV RNA, and 10 percent were positive for GBV-C E2 antibody. Results of CMV, parvovirus B19, EBV VCA, and HAV testing were similar to published background rates. No demographic or exposure history variables had significant correlation with ALT or other testing results. CONCLUSION: These data suggest that an ALT > or =120 IU per L in blood donors with negative standard screening tests has questionable value as a surrogate marker for seronegative HBV or HCV infection. Continued ALT testing may contribute little, if anything, to the safety of blood components or plasma for further manufacture. PMID- 11399815 TI - Sustained decreases in platelet count associated with multiple, regular plateletpheresis donations. AB - BACKGROUND: Transient but significant decreases in platelet counts have been documented to occur in donors undergoing single and serial short-term plateletpheresis collections. The effect of long-term regular plateletpheresis on donor platelet counts has not been characterized. STUDY DESIGN AND METHODS: A retrospective study was performed to evaluate the effects of long-term regular plateletpheresis donation on donor platelet counts. A computerized database containing records of 11,464 apheresis collections from 939 donors over a 4-year period was queried for serial preapheresis platelet counts. Donors were categorized by sex, age, and cumulative number of donations. The average difference in platelet counts (mDeltaPC) between each donor's first and last platelet count during this period was calculated for each category. A subgroup of frequent donors was selected for analysis of mDeltaPC, using the baseline platelet count obtained before the first plateletpheresis procedure. RESULTS: A significant and sustained decrease in platelet count was identified for all donation frequency categories. The frequency of donation correlated directly with decrease in platelet count for all but the highest-frequency donation group, in which deferrals for low platelet count blunted the extent of the mDeltaPC. A mean decrease of 40,000 per microL from baseline occurred in the frequent-donor subgroup. A total of 84 donors (9%) were deferred for low platelet count. The majority returned to donate successfully after temporary deferral. CONCLUSIONS: Regular plateletpheresis donors develop sustained decreases in platelet count. However, clinically significant thrombocytopenia is unusual when rigorous ongoing review and prudent deferral policies are established and followed. PMID- 11399816 TI - Platelet-specific antibodies in HLA-immunized patients receiving chronic platelet support. AB - BACKGROUND: In HLA-alloimmunized patients, the unexpected failure of HLA-matched platelet transfusions usually raises the suspicion about concomitant platelet specific antibodies. As the reported frequency of platelet-specific antibodies in multitransfused patients varies widely, the aim of this study was to determine the prevalence of such antibodies in a population of chronic thrombocytopenic patients with HLA antibodies. STUDY DESIGN AND METHODS: From 1985 to 1997, 11,777 determinations of HLA antibodies were performed in 1330 hematologic patients receiving chronic platelet support. Fifty-two patients with HLA alloimmunization that lasted more than 1 month were selected. The search for platelet-specific antibodies was performed by using a monoclonal antibody immobilization of platelet antigens assay, thus allowing the identification of platelet-specific antibodies directed against the platelet glycoproteins (GP) Ib/IX, GPIIb/IIIa, and GPIa/IIa. Specificity of the platelet-specific antibodies was further investigated by using a solid-phase assay with chloroquine-treated platelets. RESULTS: Only 2 (3.8%) of the 52 patients had platelet-specific antibodies. One antibody reacted with an epitope of the GPIIb/IIIa that was present in all the panel platelets, and that probably was an autoantibody. The other was an anti-HPA 5b. CONCLUSIONS: The prevalence of platelet-specific antibodies in patients with HLA alloimmunization is very small. The search for concomitant platelet-specific antibodies would be indicated only when other causes of refractoriness to HLA matched platelets are ruled out. PMID- 11399817 TI - Platelet alloantibodies in transfused patients. AB - BACKGROUND: Patients receiving cellular blood components may form HLA antibodies and platelet-specific alloantibodies. STUDY DESIGN AND METHODS: Serum samples from a cohort of 252 patients with hematologic or oncologic diseases who are receiving cellular blood components were studied for platelet-reactive antibodies. Specificity of platelet alloantibodies was determined with a panel of typed platelets RESULTS: Platelet-reactive antibodies were detected in the sera of 113 patients (44.8% of 252), HLA antibodies in the sera of 108 (42.9%), and platelet-specific antibodies in the sera of 20 (8%). The following platelet specific antibodies were identified: anti-HPA-5b (n = 10), anti-HPA-1b (n = 4), anti-HPA-5a (n = 2), anti-HPA-1a (n = 1), anti-HPA-2b (n = 1), anti-HPA-1b+5b (n = 1), and anti-HPA-1b+2b (n = 1). Fifteen sera from the 108 patients with anti HLA (13.9%) contained additional platelet-specific alloantibodies, while in 5 sera, platelet-specific alloantibodies only were detected: anti-HPA-5b (n = 4) and anti-HPA-1a (n = 1). Of the 108 sera with HLA antibodies, 29 (26.9%) showed discordant results when studied with the lymphocytotoxicity test and the glycoprotein-specific immunoassay. Ten sera contained panreactive antibodies against platelet glycoproteins (GP) IIb/IIIa, GPIa/IIa, and/or GPIb/IX. Alloimmunization occurred in 58.3 percent of female patients with previous pregnancies, but in only 23.3 percent of those without previous pregnancies (p = 0.0049). CONCLUSION: Platelet alloantibody specificities in transfused patients (predominantly anti-HPA-5b and -1b with antigen frequencies <30% among whites) differ significantly from those observed in patients with neonatal alloimmune thrombocytopenia or posttransfusion purpura, in whom anti-HPA-1a (antigen frequency >95%) is the most prevalent specificity. HLA antibody detection yields discordant results when the lymphocytotoxicity assay and a glycoprotein-specific immunoglobulin-binding assay are used. PMID- 11399818 TI - Platelet-reactive HLA antibodies associated with low posttransfusion platelet increments:a comparison between the monoclonal antibody-specific immobilization of platelet antigens assay and the lymphocytotoxicity test. AB - BACKGROUND: Platelet-reactive HLA antibodies are a major reason for low posttransfusion platelet increments. The clinical importance and value of the test systems for their in vitro determination is still controversial. STUDY DESIGN AND METHODS: A prospective analysis of HLA antibodies was performed in sera obtained once a week for at least 4 consecutive weeks from 55 patients (female/male, 28/27; age: median, 49 years; range, 18-69) undergoing intensive chemotherapy and in need of prophylactic platelet transfusions. All sera (n = 330) were analyzed by the monoclonal antibody-specific immobilization of platelet antigens (MAIPA) assay and by the standard lymphocytotoxicity test (LCT). RESULTS: In the MAIPA, 24.5 percent of sera (81/330) obtained from 22 patients contained HLA antibodies. These were detected significantly more often by the MAIPA assay than by the LCT (24.5% vs. 8.2%). Fifty-five sera (20 patients) were positive in the MAIPA assay only. In 15 patients, HLA antibodies were transient. In 3 patients, HLA antibodies were detected earlier by the MAIPA assay than by the LCT. Significantly more sera obtained at the time of low posttransfusion platelet increments were positive in MAIPA alone, rather than in both MAIPA and the LCT (44% vs. 17%). CONCLUSION: The MAIPA assay is more sensitive than the standard LCT in detecting platelet-reactive HLA antibodies. These MAIPA positive/LCT-negative HLA antibodies affect the posttransfusion platelet increment. PMID- 11399819 TI - The expression of NA antigens in people with unusual Fcgamma receptor III genotypes. AB - BACKGROUND: The Fcgamma receptor IIIb (FcgammaRIIIB) genes that encode neutrophil specific antigens NA1 and NA2 differ at 5 nucleotides (nts); in 4, the result is an amino acid (AA) difference between the two alleles. The role of each of these differences in antigen expression is not known. Persons with FcgammaRIIIB genes that differ from NA1-FcgammaRIIIB and NA2-FcgammaRIIIB by 1 nt have been described. This study compared NA1 and NA2 expression on granulocytes in persons with variant FcgammaRIIIB genes and in healthy blood donors. STUDY DESIGN AND METHODS: Reactions of NA1- and NA2-specific MoAbs and alloantibodies with granulocytes were assessed by flow cytometry in 74 healthy blood donors and 6 persons with known variant FcgammaRIIIB genes. The granulocytes were tested with 1 NA1-specific MoAb, 1 NA2-specific MoAb, 4 NA1-specific alloantibodies, and 4 NA2-specific alloantibodies. RESULTS: Analysis of granulocytes from persons with variant NA genotypes found that single-base substitutions in FcgammaRIIIB at 141 and at 349 are important in NA1 expression and those at 227 and 277 are important in NA2 expression. Among blood donors, neither age, sex, nor race affected the expression of NA1 or NA2. The NA2-specific MoAb reacted more intensely with granulocytes from NA2-double-dose cells than with those from NA-single-dose cells, but this was not true for the NA2-specific alloantibodies. There was no difference in the reactions of the NA1-specific MoAbs and alloantibodies with donor samples of known NA1-double-dose or NA-single-dose cells. The intensity of reactions of both the NA1- and NA2-specific MoAbs and alloantibodies were strongly correlated on double-dose cells but not on single-dose cells. In fact, granulocytes from 7 healthy blood donors, phenotyped as NA-single-dose with the MoAbs, were phenotyped as NA2-double-dose with the alloantibodies. Variations in FcgammaRIIIB are common in blacks, but 5 of the 6 donors were white. These results suggest that FcgammaRIIIB variations may be common in both whites and blacks. CONCLUSIONS: NA2 expression is affected by polymorphisms in FcgammaRIIIB 227 and FcgammaRIIIB 277, both of which are involved in an FcgammaRIIIb N glycosylation site. Polymorphisms in FcgammaRIIIB at 141 and 349 appear more important to NA1 expression. PMID- 11399820 TI - Immune reconstitution after autologous selected peripheral blood progenitor cell transplantation: comparison of two CD34+ cell-selection systems. AB - BACKGROUND: Selection of CD34+ PBPCs has been applied as a method of reducing graft contamination from neoplastic cells. This procedure seems to delay lymphocyte recovery, while myeloid engraftment is no different from that with unselected PBPC transplants. STUDY DESIGN AND METHODS: Lymphocyte recovery was studied in two groups of patients who underwent autologous CD34+ PBPC transplant with two different technologies (Ceprate SC, Cellpro [n = 17]; CliniMACS, Miltenyi Biotech [n = 13]). The median number of CD34+ cells transfused was 3.88 x 10(6) per kg and 3.32 x 10(6) per kg, respectively. Residual CD3 cells x 10(6) per kg were 4.97 and 0.58, respectively (p = 0.041). Residual CD19 cells x 10(6) per kg were 1.33 and 0.73, respectively (NS). RESULTS: No differences were found between the two groups in total lymphocyte recovery to >0.5 x 10(9) per L, which achieved a stable count by Day 30. During the study period, the CD4+ cell count remained below 0.2 x 10(9) per L, and the B-cell subset showed a trend toward normalization. CD3/HLA-DR+ and CD16/56 increased markedly in both groups by Day 30. An increase in CMV (13%) and adenovirus (17.4%) infection was found in both groups. CONCLUSION: Both CD34+ cell selection technologies used here determined an excellent CD34+ cell purity and an optimal depletion of T cells. The high rate of viral complications is probably due to the inability of residual T cells left from the CD34+ cell selection to generate, immediately after transplant, an adequate number of virus-specific lymphocytes. PMID- 11399821 TI - Association of ABO-mismatched platelet transfusions with morbidity and mortality in cardiac surgery. AB - BACKGROUND: The transfusion of ABO-mismatched platelets has been associated with increased morbidity and mortality during induction therapy for acute leukemia and allogeneic progenitor cell transplantation. STUDY DESIGN AND METHODS: Reported here is a cohort study of 153 patients undergoing primary coronary artery bypass graft or coronary valve replacement surgery by two surgeons in one institution during 1997 and 1998. All statistics employed nonparametric two-sided tests (Mann Whitney; Fisher's exact test). RESULTS: Patients receiving at least one ABO mismatched pool of platelets had a significantly longer hospital stay, more days of fever, greater total hospital charges, and more RBC transfusions. Mortality, hours in the intensive care unit, days on antibiotics, and numbers of platelet transfusions were also greater in recipients of ABO-mismatched platelets, but these differences were of less statistical significance. When the analysis was restricted to the 139 patients who received no more than two pools of platelets, the trends for increased morbidity and mortality (8.6% vs. 1.9%; p = 0.10) in recipients of ABO-mismatched platelets persisted. The number of RBC transfusions required in this latter cohort was 50 percent greater (mean, 6.1 vs. 9.2; p = 0.02), despite the fact that the number of platelet transfusions given was similar (mean, 1.2 vs. 1.3 pools; p = 0.22). CONCLUSIONS: ABO-mismatched platelet transfusions are associated with unfavorable outcomes in cardiac surgery, a relationship that remains unexplained. As this association has been found in three cohort studies in various clinical settings, further investigation of this association is warranted. PMID- 11399822 TI - Number of RBC units and rate of transfusionto anemic HIV-positive patients assigned to receiveWBC-reduced or non-WBC-reduced RBCs: the viral activation transfusion study experience. AB - BACKGROUND: It is known that the use of filtration to reduce WBCs in RBC units is associated with a 6- to 15-percent loss of RBCs. It is not known if the use of such WBC-reduced RBCs results in an increased need for RBC units or in the transfusion of more units per year to patients with anemia. STUDY DESIGN AND METHODS: In the multicenter Viral Activation Transfusion Study (VATS), anemic HIV positive patients were randomly assigned to receive either WBC-reduced or non-WBC reduced RBCs. The number of RBC units transfused per patient and the rate of RBC use were studied. All RBC units given after the enrollment transfusion were counted, until the end of follow-up or the occurrence of bleeding (receiving >5 RBCs within 2 consecutive days). RESULTS: As expected, the WBC-reduced RBC units in VATS were lighter in weight than the non-WBC-reduced units (median weight: WBC reduced, 300 g; non-WBC-reduced, 330 g; p<0.0001). After the enrollment transfusion, 258 WBC-reduced arm patients received 1279 units of RBCs (average, 5.0 units/patient, median, 2 units) while 262 patients in the non-WBC-reduced arm received 1111 RBCs (4.2 units/patient; median, 2 units). The number of units transfused for anemia was slightly greater in the WBC-reduced arm, but the difference was not significant (p = 0.41). Similarly, the rate of RBC use was somewhat higher in the WBC-reduced arm, but the difference was not significant (p = 0.14). The median was 2.3 units per patient per year of follow-up in the WBC reduced arm; the median in the non-WBC-reduced arm was 1.2 units. CONCLUSION: This study confirms that WBC-reduced RBC units are significantly lighter in weight than non-WBC-reduced RBCs. However, in the setting of a large, randomized, blinded study of transfusion for anemia, the smaller size of the WBC-reduced RBC units had no significant effect on the number of RBC units transfused or on the rate at which RBC units were used. In this study, the frequency of blood transfusion may have had a greater relationship to the frequency of routine, scheduled appointments or transfusion orders for a specified Hb trigger than to the actual Hb content of the unit. PMID- 11399823 TI - The effect of external pressure, catheter gauge, and storage time on hemolysis in RBC transfusion. AB - BACKGROUND: RBC transfusions are often administered using externally applied pneumatic pressure. The effect of this practice on RBC hemolysis was studied during simulated transfusion performed in the laboratory. MATERIALS AND METHODS: RBC transfusions were performed sequentially via 16-, 18-, 20-, and 22-gauge cannulas with the application of a pneumatic pressure device at pressures of 150 and 300 mm Hg. Hb concentration, Hct, RBC count, free Hb, potassium, and LDH concentrations were measured in 5 mL of transfusate. RESULTS: Forty-seven RBC units, 20 fresh units (mean age, 10.8 days) and 27 old units (mean age, 28.9 days) were studied under all conditions. Multiple regression analysis revealed that the age of the unit and the external pressure applied were significant determinants of the degree of hemolysis. CONCLUSIONS: The application of an external pressure device results in minimal destruction of transfused RBCs, even under the most stringent conditions examined. Thus, external pressure application to expedite an RBC transfusion is likely to be a safe procedure for the majority of patients. Unusual clinical situations, such as massive transfusion in pediatric patients, should be specifically examined to confirm the safety of this procedure. PMID- 11399824 TI - Platelet transfusions in the neonatal intensive care unit:factors predicting which patients will require multiple transfusions. AB - BACKGROUND: Previous studies suggest that recombinant thrombopoietin (rTPO) will increase platelet production in thrombocytopenic neonates. However, the target populations of neonates most likely to benefit should be defined. Studies suggest that rTPO will not elevate the platelet count until 5 days after the start of treatment. Therefore, the neonates who might benefit from rTPO are those who will require multiple platelet transfusions for more than 5 days. This study was designed to find means of prospectively identifying these patients. STUDY DESIGN AND METHODS: A historic cohort study of all patients in the neonatal intensive care unit (NICU) at the University of Florida who received platelet transfusions from January 1, 1997, through December 31, 1998, was conducted. RESULTS: Of the 1389 patients admitted to the NICU during the study period, 131 (9.4%) received platelet transfusions. Seventeen were treated with extracorporeal membrane oxygenation and were excluded from further analysis. Of the remaining 114 patients, 55 (48%) received one transfusion and 59 (52%) received more than one transfusion (21 had >4). None of the demographic factors examined predicted multiple platelet transfusions. However, two clinical conditions did; liver disease and renal insufficiency. Neonates who received one platelet transfusion had a relative risk of death 10.4 times that in neonates who received none (p = 0.0001). Neonates who received >4 platelet transfusions had a risk of death 29.9 times that in those who received no transfusions (p = 0.0001). CONCLUSION: NICU patients with liver disease or renal insufficiency who receive one platelet transfusion are likely to receive additional transfusions. Therefore, these patients constitute a possible study population for a Phase I/II rTPO trial. PMID- 11399825 TI - The effect of the interruption of agitation on platelet quality during storage for transfusion. AB - BACKGROUND: A considerable amount of data and the CFR suggest that platelet concentrates (PCs) should be stored with continuous, gentle agitation before transfusion. However, there are only limited data concerning the mechanisms of platelet damage that may occur when agitation is interrupted, and there are no CFR guidelines concerning shipment between periods of storage. STUDY DESIGN AND METHODS: PCs were prepared by the platelet-rich plasma method and stored for 5 days at 20 to 24 degrees C; agitation was interrupted for 1 to 3 days either by simply stopping the agitator or by placing the PCs in a stationary shipping container. Measurements of platelet metabolism and quality were made during storage and on Day 5. RESULTS: With interruption on the agitator, the production of lactic acid was increased during the interruption in proportion to the number of platelets in the PC and the duration of the interruption. The pO(2) was increased during agitation interruption, which suggested a decline in oxygen utilization. With the use of the hypotonic shock response and the extent of shape change as reflections of platelet quality, there was no evidence of platelet damage unless the pH fell to or below 6.5. No PC reached this level after an interruption of agitation for only 1 day, irrespective of which day was chosen for interruption. PCs whose agitation was interrupted for 2 and 3 days were at risk of having a pH less than 6.5 if their contents were greater than 1.25 x 10(11) and 0.75 x 10(11) platelets, respectively. Interruption of agitation for 1 day in the shipping container produced results essentially identical to those produced by interruption on the agitator. CONCLUSION: Interruption of agitation of PCs for 1 day, either on the agitator or in the shipping container, produces no platelet damage measurable by these in vitro techniques. However, an interruption of agitation for 2 days can result in significant damage in some components. Further studies will be required to learn more about the mechanisms that lead to the metabolic changes described and to determine if the same generalizations apply to apheresis PCs and PCs prepared from pooled buffy coats. PMID- 11399826 TI - Determination of the degree of bacterial contamination of whole-blood collections using an automated microbe-detection system. AB - BACKGROUND: The prevalence of bacterial contamination in whole-blood collections, either with immediate sampling or sampling after overnight storage as whole blood at 20 degrees C, is determined. STUDY DESIGN AND METHODS: Whole blood was collected under blood bank conditions in special five-bag systems, allowing sampling in a closed system for culture bottles. Samples were taken within 2 hours after collection (Group 1) or after overnight storage of the whole blood at 20 degrees C (Group 2). Culture bottles were incubated for 7 days, and positive samples were entered on agar plates for confirmation and determination. RESULTS: In Group 1, 9219 units were tested; 27 units were positive with positive subculture, that is, 0.29 percent with a 95% CI of 0.19 to 0.42 percent. In Group 2, 9038 units were tested; 36 units were positive with positive subculture, that is, 0.39 percent with a 95% CI of 0.28 to 0.55 percent. No significant difference could be found between the two test groups. The majority of bacteria were either Staphylococcus (all coagulase-negative) or Propionibacterium species. CONCLUSION: For a total of 18,257 units, 0.34 percent (CI, 0.25-0.44) of whole-blood collections appeared to have bacterial contamination (mainly skin-derived). Overnight storage of whole blood at 20 degrees C did not have a significant effect on the prevalence of bacterial contamination. PMID- 11399827 TI - MCV as a guide to phlebotomy therapy for hemochromatosis. AB - BACKGROUND: A multitude of recommendations exist for laboratory assays to monitor the pace and endpoints of phlebotomy therapy for hemochromatosis. All of these recommendations rely on an assessment of storage iron to guide treatment, and none have been prospectively evaluated. STUDY DESIGN AND METHODS: Nine consecutive patients underwent serial monitoring of Hb, MCV, transferrin saturation, and ferritin during weekly phlebotomy to deplete iron stores (induction therapy) and less frequent sessions to prevent iron reaccumulation (maintenance therapy). Changes in MCV and Hb were used to guide the pace of phlebotomy over a median of 7 years of follow-up. RESULTS: During induction therapy, the MCV increased transiently because of reticulocytosis and then stabilized for a prolonged period before decreasing more sharply, which reflected iron-limited erythropoiesis. Iron depletion was achieved after a median of 38 phlebotomies and removal of 9.0 g of iron. Maintenance phlebotomy was targeted to maintain the MCV at 5 to 10 percent below prephlebotomy values and the Hb at >13 g per dL. Transferrin saturation fluctuated considerably during treatment, but remained below 35 percent during MCV-guided maintenance therapy. Ferritin values were not useful guides to the pace of phlebotomy. The median maintenance therapy phlebotomy interval was 7.5 weeks (range, 6-16), which corresponded to an average daily iron removal of 35 to 67 microg per kg. Most patients showed evidence of iron reaccumulation at phlebotomy intervals of 8 weeks or more. CONCLUSION: The MCV is an inexpensive, precise, physiologic indicator of erythropoietic iron availability. When used in conjunction with the Hb, it is a clinically useful guide to the pace of phlebotomy therapy for hemochromatosis. PMID- 11399828 TI - HFE mutations do not account for transfusional iron overload in patients with acute myeloid leukemia. AB - BACKGROUND: Hereditary hemochromatosis (HH) is a HFE gene-linked disorder affecting 1 of 200 to 400 persons in white populations. It has been proposed that patients with a hematologic malignancy who are receiving frequent RBC transfusions should be screened for HFE mutations. This would identify C282Y homozygotes, who have a high risk of developing severe iron overload. STUDY DESIGN AND METHODS: DNA samples from 128 controls and 23 adult long-term survivors of acute myeloid leukemia (AML) treated at the Oulu University Hospital (Oulu, Finland) from 1987 to 2000 were examined for the presence of the C282Y and H63D mutations in HFE. All the patients were severely iron-overloaded, as determined from high serum ferritin values and/or increased storage iron in bone marrow. Phlebotomies were performed in five patients because of the symptoms of iron overload. DNA extracted from the blood was used to amplify HFE gene fragments by the PCR method, after which the amplification products were digested with restriction endonucleases SnaB I and Bcl I, and the restriction fragments were analyzed on agarose gels. RESULTS: No chromosomes with the C282Y mutation were found among the AML patients, and 5 patients (21.7%) were heterozygous for the H63D mutation. In the control group, 13 persons (10.2%) were heterozygous for the C282Y mutation and 26 (20.3%) for the H63D mutation, including 3 C282Y/H63D double heterozygotes. CONCLUSION: HFE mutations do not account for the harmful iron overload that develops in AML patients who receive large quantities of RBC concentrates after intensive chemotherapy. PMID- 11399829 TI - Health and economic consequences of HCV lookback. AB - BACKGROUND: Several countries have conducted or are considering campaigns of lookback on blood recipients who may have acquired posttransfusion HCV (PT-HCV) before the implementation of anti-HCV screening. There is, however, no estimation of the health and economic consequences of the medical interventions triggered by the lookback. STUDY DESIGN AND METHODS: This study used a Monte Carlo simulation of a Markov model representing the natural history of PT-HCV. Unadjusted and quality-adjusted life expectancy and lifetime medical costs were calculated for a cohort of patients in whom PT-HCV is diagnosed through the lookback, and these values were compared with those calculated for a similar cohort on whom lookback is not performed. RESULTS: The model predicts that 47 percent of people who received transfusions of HCV-infective blood 10 years ago are still alive, carry the infection, and have not yet progressed to end-stage liver failure. In this population, forthcoming complications of PT-HCV will reduce the remaining life expectancy by 1.75 years per patient. Medical interventions triggered by the diagnosis of PT-HCV would salvage 0.123 years of life expectancy, at a net cost of $921 per newly diagnosed patient. The health and economic impact of diagnosing a new case of PT-HCV through lookback was sensitive to the patient's age, the efficacy of antiviral therapies, the time elapsed from transfusion to lookback, and the future inflation of costs of treating end-stage liver disease. Under some plausible assumptions, the intervention could result in net financial savings for the health care system, but it may also produce a net health loss for the majority of patients who will be said to be HCV-positive without being offered an effective therapy. CONCLUSION: Diagnosis of PT-HCV through HCV lookback has a potential both to increase patients' life expectancy and to reduce health care costs. However, more effective antiviral therapies and a better knowledge of factors predicting the progression of PT-HCV are needed to attain those goals. Meanwhile, care should be taken to avoid pursuing a health gain for a minority that might result in a health loss for the majority. PMID- 11399830 TI - Cryoprecipitate-reduced plasma:rationale for use and efficacy in the treatment of thrombotic thrombocytopenic purpura. PMID- 11399831 TI - Reducing the residual risk of transfusion-transmitted viruses: mini-pool or single-donation NAT? PMID- 11399833 TI - Study of four families concerning the linkage and inheritance of the allele encoding the granulocyte-specific antigen HNA-1c (SH). PMID- 11399834 TI - The first case of HCV seroconversion in Portugal after the introduction of HCV NAT screening. PMID- 11399837 TI - The improbable "Elephant Man". PMID- 11399838 TI - T.gondii--a reply. PMID- 11399839 TI - Foot and mouth disease--the other factors. PMID- 11399840 TI - Figs and fig wasps: evolution in a microcosm. AB - Fig wasps are tiny insects that both pollinate and feed upon fig plants. Each species requires the other in order to complete its reproductive cycle. The interaction centres on the unique structure of syconium (fig inflorescence), which provides an intriguing and convenient microcosm for studying the action of both natural and sexual selection. PMID- 11399841 TI - The origin of snakes: limits of a scientific debate. AB - The origin of snakes has recently become a palaeontological 'hot potato'. Two groups of scientists have come to very different conclusions from the studying the same fossils. Are there tests capable of rejecting wither one of these alternative conclusions or will human interpretation always allow such differences of opinion? PMID- 11399842 TI - Plant antioxidants: colour me healthy. AB - Plants make a variety of compounds in response to environmental stress, many of which function as antioxidants when consumed. The plants' own defences against oxidative stress can be used for your benefit, prolonging your life by acquiring their protection. By eating plenty of vegetables and fruit, you may help to significantly reduce the risk of many age-related degenerative diseases. PMID- 11399843 TI - Dam worms. AB - Human and animal infection rates with the Oriental schistosome have steadily declined in China over the last half-century, but the Three Gorges Dam may reverse this decline by creating new, or enlarging existing, ideal environments for the worm and its aquatic snail intermediate host. PMID- 11399844 TI - Culture collections in the twenty-first century. AB - Culture collections conserve the living tools for biotechnology. Without them there would be no reference organisms, and no stocks of crucial or rare microorganisms that are so valuable for biotechnology and biomedical research. The expertise that drives these collections is under threat, but the collections themselves may survive by pooling their knowledge. PMID- 11399845 TI - Xenotransplantation: hope or delusion? AB - Having moved from the realms of science fiction to a rapidly growing research area, xenotransplantation promises numerous benefits to patients in the future. However, deep suspicions of this technology are rife. We suggest an alternative viewpoint to that given in E M Engels' article in Biologist 46(2). PMID- 11399846 TI - A foreigner in Ireland. AB - Those beautiful blue eucalyptus leaves in your Valentine's bouquet may be the product of the new cut-foliage industry in County Kerry, Ireland. Initially, commercial plantations were the victims of an insect accidentally imported from Australia, but two years ago a natural enemy was released and has proven to be an effective control agent for the pest. PMID- 11399847 TI - Who was... Emil von Behring? AB - Emil von Behring was first in the line of distinguished immunologists to win the Nobel Prize for Physiology and Medicine. His contributions to our knowledge of immunity ignited an impassioned argument between French and German scientists at the end of the 19th century, the first of many scientific debates in the immunological world. PMID- 11399848 TI - Genetic databases. AB - A simple visit to the doctor in a few years' time might see you taking part in the largest research project ever conducted in the UK. The ethical and logistical challenges of this study are as complex as the scientific ones.' PMID- 11399849 TI - The origin, formation and developmental significance of the epicardium: a review. AB - Questions on the embryonic origin and developmental significance of the epicardium did not receive much recognition for more than a century. It was generally thought that the epicardium was derived from the outermost layer of the primitive myocardium of the early embryonic heart tube. During the past few years, however, there has been an increasing interest in the development of the epicardium. This was caused by a series of new embryological data. The first data showed that the epicardium did not derive from the primitive myocardium but from a primarily extracardiac primordium, called the proepicardial serosa. Subsequent data then suggested that the proepicardial serosa and the newly formed epicardium provided nearly all cellular elements of the subepicardial and intermyocardial connective tissue, and of the coronary vasculature. Recent data even suggest important modulatory roles of the epicardium and of other proepicardium-derived cells in the differentiation of the embryonic myocardium and cardiac conduction system. The present paper reviews our current knowledge on the origin and embryonic development of the epicardium. PMID- 11399850 TI - Cerebellar glia cells induce a correct laminar organization in chicken retinal reaggregates. AB - We investigated the functional role of glia cells during retinogenesis using the rotation culture system. Reaggregating cells from the embryonic chick retina have the unique capacity to reassemble into laminated cellular spheres. These spheres are composed of several compartments holding the constituents of many retinal layers in a topologically correct, yet inverse orientation. However, when these spheres are cultured in the presence of conditioned media derived from monolayers of cerebellar glia cells, the reassembling retinal cells behave totally differently. The anlage of the originally reversed lamina polarity is progressively transformed within a week into a sphere with a compound and correctly laminated orientation. Conditioned media from fibroblasts, other glia cells (except Muller cells) or a set of already characterized retinogenetic factors are not able to produce this dramatic transformation. Additionally, we were able to show that only retinal cells are able to respond with a reorganization process. Reaggregating cells from the chick cerebellum also form spheroids; however, neither in the presence of cerebellar glia cell-derived conditioned medium nor their control counterparts are they able to reassemble histotypically. This indicates that cerebellar glia cells produce diffusible factors to which retinal cells can respond and that these factors can act as important determinants for the correct establishment of the retinal polarity. Since all types of laminar disorganization are of great clinical significance, the knowledge of factors which determine and sustain the normal retinal architecture are biomedically highly relevant. PMID- 11399851 TI - Apoptosis in cultured rat islets of langerhans and occurrence of Bcl-2, Bak, Bax, Fas and Fas ligand. AB - Isolated Langerhans islets are widely used for diabetic transplantation experiments and investigations of the mechanisms leading to the death or survival of insulin-producing cells in cultured islets. The present study was aimed at investigating programmed cell death and the role of apoptosis-associated peptides in insulin and glucagon cells of islets isolated from untreated rats and held in cultured suspension. Islets were removed from medium on days 0, 7, 14, 21 and 29, embedded in Epon, and semi-thin serial sections were prepared. At designated intervals, histologic sections were treated with the direct fluorescein-labelled TUNEL method and immunostained for pancreatic hormones (glucagon, insulin) and apoptotic peptides [Bak, Bax, Fas, Fas ligand (FasL)], as well as for the anti apoptotic peptide Bcl-2. All tissue sections were investigated using confocal laser scanning microscopy under identical setting for semiquantitative estimation of staining intensity. The percentage of apoptotic cells was between 1.6 and 2.1% and most apoptotic cells were beta-cells. Corresponding cells often contained Bak and Bax. Fas and FasL were mostly detected in islet cells within the first week after preparing the cultured suspension. The insulin content was low (1.1 +/- 0.22 ng per islet) directly after isolation. It then increased progressively up to day 14, after which it began to decrease. Glucagon expression, on the other hand, remained high for the entire duration of the investigation. In conclusion, the islet beta-cells may recover after the isolation procedure, but after 4 weeks in culture, both the insulin content and Bcl-2 staining decrease. Moreover, apoptosis is mediated by different mechanisms after the isolation procedure and after culturing the islets for 1 month. The present data may be important for further studies on isolated, cultivated or transplanted islets. PMID- 11399852 TI - Liver regeneration on chicken chorioallantoic membrane. AB - To explore how the liver regenerates, liver pieces from 15-day-old chicken embryos were grafted onto the chorioallantoic membrane (CAM) of 9-day-old chicken embryos and cultured for 11 days at the longest. The cultured liver pieces were examined histologically. The liver implants were gradually engulfed into the CAM and underwent necrosis of hepatocytes, except in their peripheral areas, during the first 1-4 days after grafting. Surviving cells in the peripheral areas began to proliferate 4 days after grafting. Thereafter, the cells were assembled into normal liver tissues and represented almost all the areas of the implants 9 days after grafting. Only after penetration of blood vessels from CAM did the liver implants enter a phase of rapid regeneration to form well-organized liver tissues. At the early stage of regeneration, the cells at the peripheral areas did not produce albumin, but reproduced it in the regenerated liver tissues, implying that hepatocytes restored their functions that were temporarily lost in the process of regeneration. Thus, we concluded that the liver pieces from 15-day old chicken embryos had the ability to form normal liver tissues on CAM and that the blood supply played an important role in liver regeneration. PMID- 11399853 TI - Chemically defined protein-free in vitro culture of mammalian embryo does not restrict its developmental potential for differentiation of skin appendages. AB - In a unique serum- and protein-free chemically defined in vitro culture model of postimplantation mammalian development the epidermis differentiates regularly, although the differentiation of other tissues is impaired due to the lack of the serum. The present study in that model was done to estimate more carefully the degree of epidermal differentiation in defined media supplemented with some growth- or differentiation-stimulating substances. The main objective was to discover by grafting in vivo to the richer environment whether simple protein free culture conditions restrict an inherent embryonic potential for differentiation of skin appendages. Embryonic parts of E9.5 gastrulating Fischer rat embryos were cultivated for 2 weeks in the protein-free Eagle's minimum essential medium supplemented with holotransferrin, apotransferrin, insulin and/or Na(2)SeO(3) and in controls cultivated in protein-free medium or in serum supplemented medium. In all experiments there was a high incidence of differentiation of the epidermis. A high level of epidermal differentiation was confirmed for the first time at the ultrastructural level. A well-differentiated cornified layer and cells connected with desmosomes containing keratohyaline masses and cytokeratin filaments were found. A strong immunohistochemical signal for the proliferating cell nuclear antigen was always detected in the basal layer of the epidermis showing that those cells were still able to proliferate. Finally, embryos precultivated for 1 or 2 weeks in the protein-free medium and media supplemented with apotransferrin or serum were grafted under the kidney capsule for an additional 2 weeks. It was discovered that even after spending 2 weeks in the simple protein-free medium in vitro, embryos retained their developmental potential for differentiation of skin appendages (hair and sebaceous glands). PMID- 11399854 TI - Degeneration and regeneration of the lip mucosal epithelium after cryo treatment in mice. AB - The process of degeneration and regeneration of the lip mucosal epithelium after cryo treatment was observed by transmission electron microscopy. The epithelial cells were degenerated by the formation of ice crystals and subsequently detached from the basement membrane, forming a blister cavity. The separation occurred between the epithelial cells and the lamina densa, leaving a small amount of cell debris on the lamina densa. The surviving cells at the periphery of the blister cavity, especially the cells in the basal half of the epithelium, provided the regeneration cells. They migrated over the cell debris, attached to the lamina densa and gradually phagocytozed it. Finally, they formed hemidesmosomes with the old lamina densa. The connections between the epithelial cells by desmosomes were so tight that desmosomes were preserved even between dead cells and between dead and living cells. Regenerating cells were moving in a multilayered form, remaining connected to each other by the dosmosomes. They were seen to divide by mitosis and thereby increase the number of the cell layer, whilst maintaining their connections with the neighbouring cells. PMID- 11399855 TI - Three-dimensional reconstruction of the anal striated musculature in a human fetus. AB - Conflicting opinions in the recent literature indicate that the morphological organization and function of the anorectal continence organ has, up to now, not been clearly understood. But a clear imagination of the spatial arrangements of this compound muscle system is of clinical relevance for the pediatric surgeon performing reconstructive surgery. We analyzed 18-microm sections of the pelvic region of 4 human fetuses in order to describe the individual components of this muscle complex. A series of 630 Azan-stained sections was the base for the computer-assisted 3D reconstruction of the levator ani and the external sphincter complex in a male human fetus (14th week p.c.). In this context, special attention was paid to the intermediate muscle layer of the puborectalis which develops ventrally from the funnel-shaped levator ani and joins the tripartite ring system of the sphincter muscle dorsally. Our findings lead to a clear imagination of the spatial arrangement of this intermediate layer and characterize the anorectal muscle complex as an integrated ensemble in which the puborectalis holds a key position. PMID- 11399856 TI - Scanning electron-microscopic view of the oral and basal epithelial surfaces of the rat soft palate. AB - We studied corresponding structures on the oral and basal surfaces of the oral epithelial layer, focusing on the microanatomy of gustatory papillae. Specimens for scanning electron microscopy were obtained from the rat soft palates and fixed with a mixture of 2% paraformaldehyde and 1% glutaraldehyde. They were first prepared without postfixation and conductive-staining to study the oral surface. After examination, the epithelium was reinforced by additional sputter coating and treated with 6 N NaOH at 60 degrees C to exfoliate the basal epithelial surfaces without any significant artifacts. The papillae, showing circular, elliptical or fusiform protuberances on the oral surface, were classified into two types: types I and II. The type I and type II papillae contained one and two taste pores, respectively. On the basal epithelial surface, the basal portions of the taste buds were associated with concentrically arranged nerve fibers and Schwann cells. Another characteristic finding on the basal epithelial surface was the presence of excretory ducts of minor salivary glands in a close spatial relationship to taste buds. It is suggested that saliva coming out through the duct is mixed with food, thus enabling intimate contact with the taste pores of the papillae. PMID- 11399857 TI - The microanatomy of the rectal salt gland of the Port Jackson Shark, Heterodontus portusjacksoni (Meyer) (Heterodontidae): suggestions for a counter-current exchange system. AB - A comprehensive anatomical study was undertaken to examine the rectal salt gland in the Port Jackson shark, Heterodontus portusjacksoni, a shark known to invade estuarine environments. The microstructure and vascular organisation of the rectal salt gland was investigated using histological observation and scanning electron microscopy of vascular corrosion casts. Cellular specialisation was observed in the lining of the central lumen of this gland. This may indicate that there is some modification of the principal product of the gland prior to its secretion. The rectal salt gland has a complex structure related to its function. Contrary to previous reports, the flow in secretory tubules is in the opposite direction to that of the capillaries and thus constitutes a counter-current arrangement. The similarity in the organisation of the counter-current and lobulate arrangement of salt-secreting glands through phylogenetically diverse organisms, such as sharks and birds, suggests that this arrangement is important in achieving efficient salt secretion. PMID- 11399858 TI - Investigation of oxacillin-hydrolyzing beta-lactamase in borderline methicillin resistant clinical isolates of Staphylococcus aureus. AB - BACKGROUND: Mechanisms of borderline resistance of Staphylococcus aureus to penicillinase-resistant penicillins (PRPs) may include hyperproduction of classical penicillinase and/or production of beta-lactamase hydrolyzing also PRPs. METHODS: beta-Lactamase activity of whole cells and purified enzymes was estimated spectrophotometrically and in isolated cytoplasmic membranes by bioassay with Bacillus subtilis as test strain. RESULTS: Out of 53 clinical isolates of S. aureus, 18 showed oxacillin MIC values from 0.5 to 2 microg/ml, which were reduced by sulbactam and/or clavulanic acid in the case of four isolates producing large quantities of inducible, type A beta-lactamase. Cytoplasmic membranes isolated from these strains showed oxacillin-hydrolyzing activity. One of these strains was grown also in the presence of globomycin, an antibiotic known to interfere with the anchorage of membrane lipoproteins; this treatment eliminated the oxacillin-hydrolyzing activity. CONCLUSIONS: The resistance in these strains was due to a membrane-bound lipoprotein with oxacillin-hydrolyzing activity. PMID- 11399859 TI - Is levofloxacin as active as ciprofloxacin against Pseudomonas aeruginosa? AB - The in vitro activity of levofloxacin against 300 Pseudomonas aeruginosa isolated from hospitalized patients, with the exception of those recovered in intensive care or hematology units, was compared to ofloxacin, ciprofloxacin, piperacillin, amikacin, ceftazidime and imipenem. Imipenem showed the best activity (81.6%), followed by piperacillin (80.7%). The activity of levofloxacin was equal to that of ciprofloxacin (75.3%) but was more active than ofloxacin (58.1%). Moreover, the MIC values of levofloxacin did not show any statistical difference using two different inocula. Levofloxacin shows an excellent bactericidal activity being generally within one doubling dilution of the MIC. These results were also confirmed by the time-killing studies. In conclusion, according to the in vitro activity, levofloxacin could be considered a good option for the treatment of infections sustained by Pseudomonas aeruginosa, and clinical experiments are required to corroborate our in vitro data. PMID- 11399860 TI - In vitro activity and post-antibiotic effect of quinupristin/dalfopristin (Synercid). AB - The in vitro activities of quinupristin/dalfopristin (Synercid), ampicillin, erythromycin, clarithromycin, vancomycin, teicoplanin, ciprofloxacin and tetracycline were examined and compared against 526 gram-positive bacteria. The minimal inhibitory concentrations (MICs) for quinupristin/dalfopristin against Staphylococcus aureus, including methicillin-resistant strains, were low (MIC(90) = 0.5 mg/l), and were comparable with those of vancomycin and teicoplanin. This compound was superior to the macrolides and highly active against Streptococcus pneumoniae (both penicillin-sensitive and penicillin-resistant strains), with MIC(90) = 2 mg/l. It was also active against other streptococci, with MIC(90) = 4 mg/l. However, this agent is less active against enterococci (MIC(90) = 32 mg/l). Quinupristin/dalfopristin showed high activity against gram-positive anaerobes, including Clostridium spp., Peptococcus spp. and Peptostreptococcus spp., with MIC(90) < or = 2 mg/l. Quinupristin/dalfopristin was also investigated for its post-antibiotic effect (PAE) and bactericidal kinetics against nine strains of gram-positive organisms, including staphylococci, enterococci and pneumococci. Exponentially growing (log phase) cultures were exposed to quinupristin/dalfopristin at 2 x MIC. Growth kinetics was evaluated using viable counting. The drug was uniformly bactericidal against pneumococci and staphylococci within 2 and 8 h of exposure, respectively. The killing activity against enterococci was weak; there was little or no reduction in bacterial count over 24 h of incubation. PAEs ranging from 2.13 to 3.28 h, 0.92 to 3.02 h and 1.89 to 7.07 h were produced on the tested pneumococci, staphylococci and enterococci, respectively. This study showed that quinupristin/dalfopristin is a promising agent active against gram-positive bacteria. The prolonged PAEs also suggest that the drug could be used intermittently at more widely spaced dosing intervals against gram-positive organisms. PMID- 11399861 TI - Inhibitory and bactericidal effects of telithromycin (HMR 3647, RU 56647) and five comparative antibiotics, used singly and in combination, against vancomycin resistant and vancomycin-susceptible enterococci. AB - The inhibitory and bactericidal effects of telithromycin (HMR 3647, RU 66647) were compared with those of gentamicin, ampicillin, erythromycin, azithromycin and vancomycin against 74 strains of enterococci (34 Enterococcus faecalis and 40 Enterococcus faecium) by agar dilution, broth dilution, time kill assays and postantibiotic effect (PAE). The telithromycin MIC(90) for vancomycin-sensitive (VSE) E. faecalis strains tested using the agar dilution method was 8 microg/ml. For a different group of VSE E. faecalis strains tested using the broth dilution method it was 0.06 microg/ml The telithromycin MIC(90)s for vancomycin-resistant (VRE) and VSE E. faecium strains, determined using the agar dilution method, were 4 and 8 microg/ml, respectively, while for a different set of VRE and VSE E. faecium strains tested using the broth macrodilution method, they were 32 and 16 microg/ml, respectively. Telithromycin MBC(90)s for E. faecalis were 4-6 tubes higher and for E. faecium 3-5 tubes higher, respectively, than the MIC(90)s. In time kill assays, telithromycin had bactericidal activity against only 1 of 7 E. faecium strains; for all other E. faecium and E. faecalis strains, only inhibitory activity was demonstrated. Neither synergy nor drug interference was observed when telithromycin was used in combination with ampicillin, vancomycin or gentamicin. At 10 times the MIC, the PAE of telithromycin against E. faecalis was 2.8 h, while for E. faecium it was 1.6 h. Telithromycin should be evaluated for therapy of enterococcal infections, including those caused by VRE organisms. However, because of the strain-to-strain variability in susceptibility to telithromycin, MIC determinations are important, especially for erythromycin resistant strains. PMID- 11399862 TI - In vitro effect of amikacin, imipenem, cefodizime, IFNalpha-2a alone and combinations of antibiotics with IFNalpha-2a on polymorphonuclear leukocyte function in chronic hepatitis patients. AB - The in vitro effect of amikacin (8 microg/ml), imipenem (30 microg/ml), cefodizime (10 microg/ml), interferon alpha-2a (IFNalpha-2a) (10 IU/ml) and antibiotic combinations with IFNalpha-2a on polymorphonuclear leukocyte (PMN) functions (phagocytosis and intracellular killing of Candida albicans blastospores) was investigated in chronic hepatitis B patients. Phagocytosis and candidacidal activity was not affected after pretreatment of PMNs with amikacin and imipenem (p > 0.05). Phagocytic activity was enhanced after pretreatment of PMNs with cefodizime and IFNalpha-2a compared with that of control PMNs (p < 0.05), but candidacidal activity was not affected by the same drugs (p > 0.05). Phagocytic and intracellular activity of PMNs were not affected by combinations of IFNalpha-2a and antibiotics (p > 0.05). PMID- 11399863 TI - Antituberculous activity of norfloxacin mannich bases with isatin derivatives. AB - Mannich bases of norfloxacin were synthesized by reacting them with formaldehyde and several isatin derivatives. The compounds were evaluated in vitro against Mycobacterium tuberculosis H37R(v) at 12.5 microg/ml in BACTEC 12B medium using the BACTEC radiometric system. Among the compounds tested, S-10 showed promising activity, with 100% inhibition at a concentration lower than 6.25 microg/ml. PMID- 11399864 TI - Pharmacokinetic profile of the anti-sickling hydroxyurea in wild-type and transgenic sickle cell mice. AB - The pharmacokinetic profile of hydroxyurea (HU) was investigated by measuring the rate of drug disappearance from the plasma in wild-type and transgenic (Tg) sickle cell mice. The absorption and elimination processes of HU exhibited first order kinetics after intraperitoneal administration of HU at 10, 50, 100 or 200 mg/kg body weight (BW). The dosage had a marked effect on the pharmacokinetics of HU in the Tg sickle cell mice. Although the area under the plasma concentration curve (AUC) increased in direct proportion with the HU dose in the wild-type mice, the AUC increased to a much greater extent at higher doses in the Tg sickle cell mice. In the Tg sickle cell mice, there was a considerable increase in the mean residence time (MRT) and a significant reduction in the apparent clearance (CL/F) at HU dose > or =100 mg/kg BW, when compared to the lower doses. At an HU dose of 200 mg/kg BW, the CL/F in the Tg sickle cell mice was reduced by about 50% of the value obtained at a dose of 10 mg/kg BW. This phenomenon was not noticeable in the wild-type mice. The MRT value in the wild-type mice at all doses was relatively constant. The steady-state distribution volume of HU in both the wild-type and Tg sickle cell mice was relatively constant at all doses of the drug. The AUC, CL/F, MRT, and terminal half-life values at any given HU dose showed significant differences between the wild-type and Tg sickle cell mice. Following intraperitoneal administration of HU at a dose of 10, 50, 100, or 200 mg/kg BW, the mean percentage of HU excreted in the urine of the wild-type and Tg sickle cell mice over 120 min was 84 +/- 6.4% and 50 +/- 8.2%, respectively, indicating a significant difference in the amount of HU excreted in urine in the two kinds of mice. The results obtained in this study may be useful in establishing an optimal dose of HU in the treatment and management of patients with sickle cell disease and other hemoglobinopathies. PMID- 11399865 TI - Evaluation of polyene-azole antagonism in liquid cultures of Candida albicans using an automated turbidometric method. AB - BACKGROUND: A computerized machine, SPECTRAmax 340, was used to evaluate the recently reported phenomenon of antagonism of the polyene amphotericin B (AMB) in Candida albicans pre-exposed to the triazole fluconazole (FLZ). METHODS: We investigated growth inhibition by varying concentrations of AMB in seven isolates of C. albicans pre-exposed to FLZ (50 microg/ml) for 18 h. All isolates were obtained on sequential visits from human immunodeficiency virus-infected patients not treated with FLZ. RESULTS: Antagonism of AMB activity was observed in 5, 4, 2 and a single isolate for 0.5, 1, 2 and 3 microg/ml of the antifungal, respectively. In the majority of Candida isolates, antagonism was seen within a concentration range of 0.5-1.0 microg/ml AMB; 1 Candida strain (HK1-Sa) was resistant to 3 microg/ml AMB. Higher concentrations of AMB (>3 microg/ml) killed both the controls and FLZ-pre-exposed Candida cells. No significant differences were observed between the periods of antagonism observed for any of the sequential isolates or for any of the AMB concentrations or in the maxima of the growth curves obtained for all Candida isolates. CONCLUSION: We conclude that the SPECTRAmax system is a useful tool to evaluate in vitro pharmacodynamic interactions between antifungal regimens within a fluid culture system, and provides information that cannot be obtained using traditional plate assay systems. PMID- 11399866 TI - Piperacillin with and without tazobactam against extended-spectrum beta-lactamase producing Pseudomonas aeruginosa in a rat thigh abscess model. AB - BACKGROUND: We compared the antibacterial effect of piperacillin, piperacillin tazobactam and imipenem in a paired rat thigh abscess model. METHODS: Two abscesses were provoked in the thighs of rats, one on the right with an extended spectrum beta-lactamase (OXA-14)-producing Pseudomonas aeruginosa (Ps-162) and the other on the left thigh with a control strain. RESULTS: The colony counts from the abscesses in log 10 colony-forming units per gram (mean +/- SD) in the imipenem group were 2.78 +/- 1.71 and 3.19 +/- 1.66, in the piperacillin tazobactam group 4.36 +/- 0.23 and 2.44 +/- 1.97, and in the piperacillin group 4.44 +/- 0.21 and 3.71 +/- 0.99 for Ps-162 and the control strain, respectively. The mean colony counts were significantly different (p < 0.05) between Ps-162 and the control strain in the piperacillin and piperacillin-tazobactam groups. CONCLUSION: These data showed that piperacillin and piperacillin-tazobactam were significantly less effective against extended-spectrum beta-lactamase-producing P. aeruginosa, while imipenem was equally effective against both Ps-162 and the control strain in this abscess model. PMID- 11399867 TI - Increased sensitivity to cisplatin in gastric cancer by antisense inhibition of the her-2/neu (c-erbB-2) gene. AB - BACKGROUND: The c-erbB-2 oncogene encodes a transmembrane tyrosine kinase receptor and its abnormal expression may be related to the prognosis of gastric cancer. Gastric cancer is relatively resistant to various drugs, including cisplatin. Cisplatin is widely used in cancer chemotherapy, but the mechanisms of drug resistance are not yet known. METHODS: We used the human gastric cancer cell lines MKN-7 and KATO-III, which express the c-erbB-2 oncogene, as a model for relative resistance to cisplatin. We investigated whether inhibition with antisense oligonucleotides against c-erbB-2 increased the sensitivity of MKN-7 and KATO-III cells to cisplatin. RESULTS: Antisense oligonucleotides for c-erbB-2 inhibited the expression of c-erbB-2 mRNA and protein and increased sensitivity to cisplatin, but not to other drugs, in MKN-7 and KATO-III cells. Cell growth was also inhibited by c-erbB-2 antisense oligonucleotides but not sense oligonucleotides. CONCLUSION: These findings indicate that c-erbB-2 expression in gastric cancer is one of the factors related to cisplatin sensitivity, and that anti-c-erbB-2 antisense oligonucleotides induced increased sensitivity to cisplatin. PMID- 11399868 TI - Evaluation of spontaneous contamination of ocular medications. AB - BACKGROUND: In order to evaluate whether single-dose ophthalmic preparations in 0.5-ml containers can safely be used within 24 h after the first opening, eigth different sterile ocular medications containing timolol, jaluronic acid, diclofenac, ketotifen, pilocarpine, formocortal, formocortal-gentamycin, and tetryzoline-feniramine (Farmigea, Italy) were opened and tested for spontaneous bacterial contamination after exposure to air. METHODS: Samples (10 microl) were collected from exposed ophthalmic preparations after 0, 2, 4, 8 and 24 h. RESULTS: No viable microorganisms were detected during and at the end of the evaluation period. In order to assess whether the resident or pathogenic ocular bacterial population due to repeated handling might contaminate the medications, about 10(5) cells of different species (Staphylococcus aureus, coagulase-negative staphylococci, Streptococcus pneumoniae, Streptococcus spp., Corynebacterium spp., Pseudomonas aeruginosa, Neisseria spp., Acinetobacter spp., Haemophilus influenzae, Escherichia coli and Candida albicans) were added to the containers and incubated at 37 degrees C or at room temperature. Samples were collected and the number of viable bacteria was estimated. The antibacterial effect of the ophthalmic compounds varied depending on the species considered. Tetryzoline feniramine, pilocarpine, ketotifen and formocortal-gentamycin exhibited a frank bactericidal activity (<100 survivors after 18-24 h of exposure) against the great majority of the species tested. CONCLUSION: These results indicate that the risk of spontaneous contamination of ophthalmic preparations after their first opening is low, and that all preparations tested exhibit an aspecific antibacterial activity. As a consequence, the safe usage of these ocular medications could be extended from the recommended 3 h to at least 24 h after the first usage. PMID- 11399869 TI - Mitochondrial ATP-sensitive potassium channel plays a dominant role in ischemic preconditioning of rabbit heart. AB - BACKGROUND: The ATP-sensitive potassium (K(ATP)) channel has been shown to be important in the ischemic preconditioning (IPC) response. Recently, the mitochondrial rather than the sarcolemmal K(ATP) channel has been focused on due to its energy-modulating property. Hence, this study was undertaken to elucidate the role of the mitochondrial K(ATP) channel in IPC by modulating the mitochondrial K(ATP) channel in isolated perfused rabbit hearts. METHODS: Seven hearts served as a control with no interventions. Seven hearts underwent IPC consisting of two 5-min cycles of global ischemia followed by 5 min of reperfusion. Seven hearts received the selective mitochondrial K(ATP) channel blocker 5-dehydroxydecanoate (5-HD, 100 microM) for 5 min before IPC, and 7 hearts received the selective mitochondrial K(ATP) channel opener diazoxide (50 microM) for 5 min. Then, all hearts were subjected to 1 h of left anterior descending coronary artery ischemia and 1 h of reperfusion. Left ventricular pressures, monophasic action potentials and coronary flow were measured throughout the experiment and infarct size was detected at the end of experiment. RESULTS: (1) The mitochondria-selective K(ATP) channel opener diazoxide reduced infarct size as compared to control (p < 0.05); (2) IPC reduced infarct size and preserved postischemic diastolic function as compared to control (p < 0.05), and (3) the mitochondria-selective K(ATP) channel blocker 5-HD reversed these effects. CONCLUSION: The mitochondrial ATP-sensitive potassium channel may be a potential site of cardioprotection. PMID- 11399870 TI - Evaluation of subcutaneous tissue gases and pH during induction of acidosis and alkalosis. An experimental study in pigs. AB - Peripheral tissue oxygen utilization was studied during hypoxic-induced acidosis and sodium bicarbonate-induced alkalosis in 8 domestic pigs by measurements of subcutaneous oxygen tension (PscO2), carbon dioxide tension (PscCO2) and pH (pH(sc)) in relation to central hemodynamic parameters and oxygenation. Hypoxic induced acidosis resulted in a decrease in P(sc)O(2) [corrected] and arterial oxygen tension (P(a)O(2)) to one third of baseline values (p < 0.05), an increase in PscCO2 and arterial carbon dioxide tension (PaCO2) from 41 to 55 and 34 to 39 mm Hg, respectively (p < 0.05), and a decrease in pH(sc) from 7.47 to 7.30 (p < 0.05). PscO2 and PaO2 increased during reversal of hypoxia and infusion of bicarbonate (p < 0.05), without reaching baseline values. In parallel PscCO2 decreased and pH(sc) increased but changes lagged behind changes in blood gases. Alkalosis established by further infusion of bicarbonate resulted in a decrease in PaO2 to 62 mm Hg whereas PscO2 remained below baseline values (p < 0.05). Correction of oxygen utilization in the subcutaneous tissue as measured by the markers PscCO2 and pH(sc) is slower than indicated by changes in tissue oxygen tension, blood gases and pH. Overcompensation of acidosis with bicarbonate resulting in alkalosis impairs oxygenation. PMID- 11399871 TI - Effect of CO2 pneumoperitoneum on the systemic and peritoneal cytokine response in a LPS-induced sepsis model. AB - We studied the effect of carbon dioxide (CO2) pneumoperitoneum on the systemic and peritoneal cytokine response in a rat model of intraperitoneal sepsis. After intraperitoneal injection of bacterial lipopolysaccharide (LPS, 10 mg/kg), rats were divided into 3 groups (n = 49 in each group): control (abdominal puncture); CO2 pneumoperitoneum, and laparotomy. Blood and peritoneal lavage fluid (PLF) were sampled at 0, 1, 2, 3, 4, 6, and 8 h after LPS challenge. Blood cell counts, plasma endotoxin level, and the levels of tumor necrosis factor-alpha (TNF alpha), interleukin-1beta (IL-1beta), and interleukin-6 (IL-6) in the plasma and PLF were measured. Blood cell counts did not differ between the 3 groups. Plasma endotoxin levels in the pneumoperitoneum group were significantly increased immediately after the procedure (p < 0.05). Although peak plasma TNF-alpha levels in the pneumoperitoneum group were seen immediately after the procedure, other changes in plasma cytokine levels did not differ significantly between the 3 groups. PLF TNF-alpha and IL-1beta levels in the pneumoperitoneum group were significantly lower than levels in the control and laparotomy groups soon after the procedure (p < 0.05). PLF IL-6 levels in the pneumoperitoneum group tended to be lower than those in the laparotomy group. In conclusion, CO2 pneumoperitoneum might induce different responses between systemic and peritoneal cytokines soon after the procedure in a rat model of intraperitoneal sepsis. PMID- 11399872 TI - Cecal ligation and puncture as a model of sepsis in the rat: influence of the puncture size on mortality, bacteremia, endotoxemia and tumor necrosis factor alpha levels. AB - BACKGROUND: Cecal ligation and puncture is a widely used experimental model of sepsis. AIM OF THE STUDY: The present study was aimed to evaluate the influence of the size of the cecal puncture on mortality, bacteremia, endotoxemia and plasma TNF-alpha levels. MATERIALS AND METHODS: Female Sprague-Dawley rats underwent cecal ligation and puncture, divided into the following groups, defined by the diameter of the cecal puncture: 0.5-cm blade incision (n = 25), 13-gauge (n = 25), 16-gauge (n = 25), 18-gauge puncture (n = 25) and 4 punctures with a 22 gauge needle (n = 25). A sham operation was performed in another 25 rats. Three animals of each group were sacrificed 5 h after the procedure for blood cultures as well as determination of plasma endotoxin and TNF-alpha. The remaining animals were followed up for a week after cecal ligation and puncture for evaluation of mortality. RESULTS: Five hours after cecal ligation and puncture, bacteremia was present in all animals, independently of the puncture size. Endotoxemia and plasma TNF levels tended to increase along with the diameter of the cecal puncture. Mortality gradually increased with the puncture size, from 27% with a 22-gauge needle to 95% with the blade incision. CONCLUSIONS: The severity of sepsis obtained with cecal ligation and puncture in rats can be easily modulated varying the size of the puncture. PMID- 11399873 TI - Does the administration route of leucovorin have any influence on the impairment of colonic healing caused by intraperitoneal 5-fluorouracil treatment? AB - Intraperitoneal chemotherapy with 5-fluorouracil (5-FU) is a new, promising alternative in adjuvant treatment of advanced colorectal cancer. Leucovorin (LV), a biomodulator of 5-FU, potentiates the antineoplastic effect of 5-FU. The aim of this study was to determine whether the administration routes of LV had any influence on the impairment of colonic healing caused by intraperitoneal 5-FU treatment. 48 male Wistar rats were subjected to left colonic resection and anastomosis, and randomized to 1 of 4 groups: control group (receiving intraperitoneal NaCl, intravenous NaCl); ipFU group (receiving intraperitoneal 5 FU, intravenous NaCl); ipFU+ivLV group (receiving intraperitoneal 5-FU, intravenous LV), and ipFU+LV group (receiving intraperitoneal 5-FU+LV, intravenous NaCl). Treatment was started after surgery and continued for 5 days with daily injections. The animals were sacrificed on the 7th day postoperatively. Anastomotic complications were more common in the ipFU, ipFU+ivLV, and ipFU+LV groups (p < 0.05) compared to the control group. The anastomotic breaking strength was significantly reduced in the ipFU, ipFU+ivLV, ipFU+LV groups (p < 0.05) than in the control group, but it did not differ between the ipFU, ipFU+ivLV, and ipFU+LV groups. The hydroxyproline content of the anastomotic segment was also significantly reduced in the ipFU, ipFU+ivLV and ipFU+LV groups (p < 0.05) compared to the control group. However, there was no difference between the anastomotic hydroxyproline content of the ipFU, ipFU+ivLV, and ipFU+LV groups. In this experiment, colonic healing was impaired after intraperitoneal 5-FU administration as judged by the higher rates of anastomotic complications, reductions in anastomotic breaking strength and hydroxyproline content; but LV administration either intravenously or intraperitoneally did not cause further deterioration in colonic healing. PMID- 11399874 TI - Effect of selective denervation of the rat pancreas on pancreatic endocrine function. AB - The purpose of this study was to investigate the influence of selective denervation of the rat pancreas on hormone secretion and on peripheral insulin sensitivity. Thirteen rats, 7 denervated and 6 sham operated, received an intravenous glucose challenge for 30 min. The basal plasma levels of insulin, glucagon and glucose did not differ between the two groups. An augmented insulin response to glucose was detected in the denervated group, whereas the glucagon response was unaffected. Glucose tolerance was marginally improved. Twenty-four rats, 12 denervated and 12 sham operated, received a constant infusion of glucose, insulin, epinephrine and propranolol in order to inhibit the endogenous insulin release and thus evaluate insulin sensitivity. No significant change in insulin sensitivity could be detected during our experimental conditions. We conclude that selective denervation brings about an increased insulin response to glucose, probably by interrupting a catecholaminergic negative tone on the beta cell. The sympathectomized animals did not disclose any apparent changes in peripheral insulin sensitivity. PMID- 11399875 TI - Effects of zinc deficiency and corticosterone elevation on bone marrow in rats. AB - OBJECTIVE: The effects of zinc deficiency and serum corticosterone elevation, in a zinc-deficient state, on rat bone marrow were studied. MATERIALS AND METHODS: Male Sprague-Dawley rats were divided into the following four groups: zinc adequate (ZA), restricted ZA (RZA), zinc-deficient (ZD) and ZD with the injection of Mifepristone (ZD+Mife). Platelet and reticulocyte counts, levels of serum zinc and corticosterone, bone marrow cellularity and morphological alterations of marrow were examined in all rats. RESULTS: Decreased marrow cellularity, decreased numbers of peripheral platelets and reticulocytes, and increased adipocytes in the marrow were observed in ZD rats. The injection of Mifepristone improved marrow cellularity, and increased the number of platelets, while decreasing adipocytes in the marrow in ZD rats. CONCLUSION: Both zinc deficiency itself and the associated serum corticosterone elevation induced bone marrow suppression in rats. PMID- 11399876 TI - A fetal lung lesion consisting of bronchogenic cyst, bronchopulmonary sequestration, and congenital cystic adenomatoid malformation: the missing link? AB - A fetus was found to have a large left thoracic cyst on routine prenatal ultrasound at 23 weeks of gestation. This lesion caused compression of the normal left lung tissue and contralateral mediastinal shift. At 23 weeks of gestation the cyst was percutaneously aspirated without subsequent reaccumulation of fluid. Serial ultrasounds showed decrease in the size of the cyst. The clinical diagnosis of congenital cystic adenomatoid malformation was made. At birth, the child had no respiratory distress, and a CT scan confirmed the finding of a fluid filled cyst in the left chest. At the time of resection, a nonaerated extralobar bronchopulmonary sequestration (with a systemic arterial blood supply and separate pleural investment) was found. The dominant cyst had ciliated respiratory epithelium with cartilage, indicative of a bronchogenic cyst, and the remainder of the specimen had the histologic hallmarks of a congenital cystic adenomatoid malformation. The coexistence of three separate anomalies in one lesion suggests a common embryological link for these malformations. PMID- 11399877 TI - Prenatal detection of facial clefts. AB - OBJECTIVES: To determine (1) the antenatal detection rate for isolated cleft lip and/or cleft palate during the routine anomaly scan; (2) the correlation between prenatal diagnosis and postnatal findings, and (3) the association of apparently isolated cleft lip and/or cleft palate with other anomalies, in particular chromosomal abnormalities. METHOD: A population-based retrospective analysis of all cases of isolated cleft lip and/or cleft during an 8-year period in an academic teaching hospital in the UK. RESULTS: Thirty-nine cases of isolated cleft lip and/or cleft palate were identified among deliveries at the hospital. Twenty-eight cases had a routine anomaly scan. Fourteen cases were detected prenatally (sensitivity 50%). None of the isolated cleft palates was detected, while 14 of 20 cases of cleft lip (70%) were detected. One of the isolated cases of cleft lip was associated with trisomy 21, while 3 of the isolated cleft palate cases were associated with the Pierre Robin syndrome. In all cases, an antenatal diagnosis of cleft was confirmed following delivery or post-mortem examination (specificity 100%). CONCLUSIONS: Ultrasound is a useful tool in screening for cleft lip with or without cleft palate, but not for cleft palate alone. Even with an isolated cleft lip, there is an increased risk of chromosomal abnormality. The role of prenatal education and support is extremely important in the preparation of prospective parents and can help alleviate the shock which occurs when there is an unexpected cleft at birth. PMID- 11399878 TI - Prenatal sonographic features of congenital lobar emphysema. AB - The prenatal sonographic features of congenital lobar emphysema (CLE) have not been well characterised. Five cases have been reported in the literature and on all these occasions either an echogenic (3) or a cystic (2) lung lesion was detected prenatally and the diagnosis was confirmed after the operation. This is the sixth case of CLE in the literature with prenatal sonographic features documented. The prenatal scans of a 23-year-old lady performed at 22 weeks of gestation revealed cystic lesions and increased echogenicity of the right fetal lung. There were no other anomalies and the karyotype was normal. The lesion decreased in size at 28 weeks and the baby was born by a normal vaginal delivery at 41 weeks. CT scan performed on day 6 confirmed cystic changes on the right lung with compression of the right lower lobe. A repeat CT scan performed at 4 months revealed extensive cystic changes in a hyper-inflated right lung and mediastinal shift to the left. At operation, abnormally inflated right upper and middle lobes were found suggesting a CLE. There were no subsequent complications after removal and histology confirmed CLE. The reported cases are reviewed and the prenatal sonographic features of CLE are discussed. PMID- 11399879 TI - Effects of intra-amniotic meconium exposure on the fetal rat: development of a pathogenic model. AB - OBJECTIVE: To develop an in vivo animal model for the study of the effects of intrauterine meconium exposure on the fetus. METHODS: Timed pregnant Long-Evans rats were purchased on gestational day (GD) 12 and allowed to acclimate for at least 48 h prior to surgery. Laparotomy was performed and both uterine horns were exteriorized through the abdominal incision. A 26-gauge needle was used to inject either 0.1-cm(3) sterile normal saline or a 20% meconium suspension into each individual gestational sac. The uterus was returned to the abdomen and the incision was closed. On GD 21 (term = 21 days) a cesarean section was completed and the number and viability of fetuses in each horn were recorded. RESULTS: A total of 14 animals were involved in this pilot study. One rat underwent sham surgery with only intra-amniotic saline injection and 13/15 fetuses survived to term. Two animals that underwent surgery on day 18 expired < 24 h postinjection. Eleven maternal animals were injected on GD 20 and underwent cesarean delivery at term; survival rates for saline-injected animals were 71.2% compared to 66.2% for meconium-exposed fetuses. CONCLUSION: We have established an in vivo animal model that allows for the examination of the effects of prolonged intrauterine meconium exposure on the fetus. PMID- 11399880 TI - Meconium staining and meconium aspiration syndrome. Is there seasonal variation? AB - OBJECTIVE: To determine whether the incidence of pregnancies complicated by meconium-stained amniotic fluid (MSAF) or meconium aspiration syndrome (MAS) differs with seasonal changes. METHODS: An established perinatal database was used to identify all term (> or = 37 weeks) singleton gestations resulting in a live birth from January 1, 1997 to December 31, 1999. Patients were divided into groups based on the season of delivery: winter (December-February), spring (March May), summer (June-August), and fall (September-November). Rates of MSAF (%MSAF/total deliveries) and MAS (%MAS/total deliveries) were calculated and compared among seasons. Local climatic data (average monthly temperature and monthly precipitation) were obtained from the National Weather Service. Multiple logistic regression analysis was performed to control for the effects of confounding variables and odds ratio (OR) with 95% confidence intervals (CI) were calculated. p < 0.05 was considered significant. RESULTS: Over the 3-year study period there were a total of 14,888 deliveries meeting the criteria. MSAF occurred in 3,206 (21.5%) deliveries and MAS developed in 92 (0.6% of total, 2.9% of MSAF). There were no differences in the rate of MSAF (p = 0.2) or MAS (p = 0.6) between seasons. By logistic regression neither season, temperature, nor precipitation were associated with MSAF or MAS. CONCLUSIONS: Our findings suggest that over the period examined there were no significant seasonal variations in the incidence of MSAF or MAS. PMID- 11399881 TI - Increased nuchal translucency, hydrops fetalis or hygroma colli. A new test strategy for early fetal aneuploidy detection. AB - OBJECTIVES: Nuchal translucency measurement of 3 mm or more (> or = 95th centile for gestation age), hydrops fetalis or hygroma colli between the 11th and 14th weeks of gestation is associated with a higher risk of fetal Down syndrome and other aneuploidies. So far, chromosome preparation of chorionic villi samplings (CVS) after short-term (or direct) culture is the only valid, reliable and rapid method of choice for the early detection of chromosomal aberrations. However, because of the placental mosaicisms detected after short-term culture, CVS has to be confirmed by a second method. Moreover, short-term villi preparation does not always provide a sufficient quantity and quality of metaphases to enable cytogenetic analysis. Unfortunately, a predicative cytogenetic result will be available only after long-term cultivation (usually after 1-2 weeks). An alternative rapid method, inexpensive and suitable for diagnosing autosomal trisomies, is the quantitative fluorescence polymerase reaction (QF-PCR) using different polymorphic small tandem repeats (STRs) on CVS-DNA. Therefore, it was the aim of the study to evaluate whether a new CVS test strategy could be employed in early pregnancies at high risk after the rapid detection of fetal chromosomal abnormalities by QF-PCR for chromosomes 13, 18 or 21 and sexing in conjunction with short-term chromosome analysis. MATERIALS: Nineteen CVS were chosen for QF-PCR detection of trisomy 21, 18 or 13 after an increased nuchal translucency measurement (> or = 95th centile for gestation age), a hydrops fetalis or a hygroma colli. The amelogenin locus of chromosomes X and Y (AMXY) were used for sexing. The QF-PCR results were compared with routine karyotyping after short- and/or long-term cultivation of CVS cells. RESULTS: An informative result was demonstrated in all analysed specimens. Nine CVS were diagnosed as a QF-PCR trisomy either for chromosome 21, 18 and 13. The pathological samples also included 4 cases of mosaicism where the normal cell line was not identified by QF PCR. In 1 additional case with a normal QF-PCR result, short-term CVS chromosome analysis showed a mosaic trisomy 13, whereas longterm CVS culture revealed a normal karyotype. The malformed aborted fetus showed no clinical signs of trisomy 13, confirming the normal results obtained by QF-PCR and long-term CVS chromosome analysis. One pregnancy with a Turner syndrome was not identified by molecular analysis. CONCLUSIONS: This study showed that all early pregnancies with a clinically relevant autosomal trisomy could be detected prenatally in routine practice by QF-PCR. The combined use of both rapid methods - QF-PCR and short term chromosome analysis - optimise the results by minimising the possibility of false-positive or false-negative findings. We believe that after verification of a pathological result obtained by two independent methods (QF-PCR and short-term CVS chromosome analysis), long-term villi cultivation is no longer necessary. However, in all cases with discrepancies, especially in samples with mosaic findings at short-term CVS cultivation, further studies are still necessary. PMID- 11399882 TI - A fetal Wolff-Parkinson-White syndrome diagnosed prenatally by magnetocardiography. AB - We report a case of fetal Wolff-Parkinson-White (WPW) syndrome diagnosed prenatally by magnetocardiography (MCG). At 32 weeks' gestation, the fetus was diagnosed to have a paroxysmal supraventricular tachycardia by ultrasonography and direct fetal electrocardiogram (ECG). Transplacental fetal therapy by maternal oral administration of propranolol resolved the fetal tachyarrhythmia. Although the wave forms of the fetal MCG at 32 weeks' gestation were normal, the fetal MCG at 35 weeks' gestation showed a short PR interval and a long QRS complex duration with a delta wave, indicating WPW syndrome. The findings of the fetal MCG were confirmed by the postnatal ECG. MCG made the prenatal diagnosis of WPW syndrome possible. PMID- 11399883 TI - Ontogenesis of villi and fetal vessels in the human placenta. AB - The ontogenesis of the villous and vascular arborescence in normal pregnancy is reviewed. The emergence of the villi and the fetal vessels is described from early pregnancy to term together with the specializations of the villi, vessels and capillaries observed in the last trimester. The expression, localization and role of the angiogenic growth factors (FGF, VEGF, PLGF, HGF) are described and discussed. Pathological pregnancies with hyper- and hypocapillarization are related to altered oxygenation. The potential roles of growth factors are presented and hypotheses proposed which may provide a molecular basis for the development of the most frequent placental pathologies. PMID- 11399884 TI - Gastroschisis complicated by midgut atresia, absorption of bowel, and closure of the abdominal wall defect. AB - We present a case of gastroschisis that was associated with progressive resorption of the extra-abdominal bowel loops and dilation of intra-abdominal bowel loops. After preterm delivery at 32 weeks, a small paraumbilical remnant was present. There was complete atresia of most of the jejunum, ileum, cecum, and the proximal half of the transverse colon. At laparotomy, the jejunum was anastomosed to the transverse colon. The neonate developed short gut syndrome and eventually received a liver and intestinal transplant. PMID- 11399885 TI - Acardiac twin with preserved brain. AB - The fatal acardiac syndrome is a rare complication of monochorionic twinning and is reported in 1 of 35,000 births. It is caused by arterioarterial and venovenous placental anastomoses leading to circulatory predominance of 1 twin. The donor 'pump' twin provides circulation for itself and for the recipient acardiac twin. The acardiac twin is usually grossly abnormal with severe reduction anomalies of the upper part of the body. We report a twin pregnancy, where a recipient twin initially by ultrasound was misdiagnosed as dead. In the third trimester the supposedly dead twin presented as an edematous acardiac twin without peripheral reduction defects and a nearly normally developed brain. An acardiac twin with a nearly normal external appearance and an almost normally developed brain, nourished by a surviving twin brother, has not previously been described in the literature. PMID- 11399886 TI - Congenital cataract in triplet pregnancy after ivf with frozen embryos: prenatal diagnosis and management. AB - Unilateral congenital cataract was diagnosed at the 2nd trimester ultrasonography in a triplet pregnancy following in vitro fertilization (with frozen embryos). Congenital cataract could be hereditary or related to metabolic and infectious disorders. To our knowledge this is the first antenatal diagnosis of the disorder in triplets after IVF with frozen embryos. PMID- 11399887 TI - Prenatal diagnosis of steroid 21-hydroxylase deficiency by allele-specific amplification. AB - OBJECTIVE: Congenital adrenal hyperplasia due to steroid 21-hydroxylase deficiency (21-OHD) is the most common cause of ambiguous genitalia in females at birth. Here, we report the first prenatal diagnosis of 21-OHD by DNA analysis in Hungary. METHODS: Allele-specific amplification (ASA) of the DNA obtained by chorionic villus sampling was performed. RESULTS: The fetus had a homozygous nonsense mutation (Gln318Stop), suggesting a salt-wasting phenotype. Dexamethasone treatment of the mother was started on the 8th gestational week and, as the fetus was an affected female, it was continued until term. The newborn had normal external genitalia at birth, and severe salt-wasting crisis and postnatal virilization was prevented by mineralo- and glucocorticoid replacement therapy. CONCLUSION: 21-OHD was genotyped by ASA, and virilization of the fetus was prevented by antenatal dexamethasone therapy. PMID- 11399888 TI - Pathogenesis of twin-twin transfusion syndrome: the renin-angiotensin system hypothesis. AB - In spite of active perinatal management, twin-twin transfusion syndrome (TTTS) remains a severe disease with a high risk of neonatal mortality and morbidity. TTTS initially results from an unbalanced blood flow from a donor to a recipient twin. However, its pathogenesis remains unclear, although cardiovascular disturbances and regulation of fetal volemia and diuresis seem central in this syndrome. Previously, we demonstrated that the renin-angiotensin system (RAS) was up-regulated in donor twins as a consequence of hypovolemia, and down-regulated in recipients. This was the first evidence of the implication of the RAS in TTTS. We hypothesize that the RAS plays a key role in the pathogenesis of TTTS. In the donor, RAS up-regulation aggravates oligohydramnios and may increase arterial resistance, which could contribute to placental dysfunction leading to intrauterine growth restriction. In the recipient, paradoxical RAS activation, due to transfer of effectors such as angiotensin II through placental shunts, could explain fetal vascular disturbances and cardiomyopathy. According to our hypothesis, TTTS would appear similar to the classical model of hypertension referred to as '2 kidneys-1 clip' with a donor twin, comparable to the clipped kidney, intoxicating its cotwin, comparable to the normal kidney. PMID- 11399889 TI - Sepsis in second trimester of pregnancy due to an infected myoma. A case report and a review of the literature. AB - Considering the high incidence of uterine myomata in women in reproductive age, myomata are only found in 2% of all pregnancies. Although they frequently lead to complications in pregnancy, cases of pyomyomata during pregnancy are rarely reported. A 44-year-old gravida 1 in her 26th week of gestation was admitted to the hospital for septic temperatures of unknown cause. A 12-cm leiomyoma with solid structures of heterogenic sonographic pattern and cystic spaces had been documented on a prior first trimester sonogram. The myoma now appeared with the same size but an increased echogenicity of the liquid parts. Ultrasound guided aspiration of the fluid within the myoma showed an infection with Klebsiella pneumoniae. A cesarian section with myo- mectomy confirmed the diagnosis of a pyomyoma. PMID- 11399890 TI - Bilateral diaphragmatic hernia followed by fetal ultrasonography. A report of two cases. AB - Two cases of bilateral congenital diaphragmatic hernia (CDH) followed by fetal ultrasonography were described. Although many cases of CDH are diagnosed by fetal ultrasonography, it is difficult to diagnose bilateral CDH in utero, which is a relatively rare and fatal condition. Two fetuses were diagnosed as having left CDH associated with severe anomalies. However, a retrospective review of fetal ultrasonography indicated elevation of the liver in the right posterior chest. Both patients died shortly after surgical repair for left CDH despite the use of extracorporeal membrane oxygenation. Diagnosis of bilateral CDH by fetal ultrasonography and the evaluation of its prognosis were discussed. PMID- 11399891 TI - A rare case of non-immune hydrops fetalis: double-chambered right ventricle. A case report. AB - A double-chambered right ventricle was diagnosed prenatally by ultrasound examination in a case of fetal hydrops and polyhydramnios. Delivery was induced at the 28th week. 2D echocardiography of the newborn confirmed the intrauterine diagnosis. To our knowledge, this is the first reported case of prenatally diagnosed divided right ventricle, causing hydrops. PMID- 11399892 TI - Possible cytotoxic effect of the expression of a connexin 43-LacZ fusion gene in cells of the vascular wall. AB - Connexin 43 (Cx43) gap junctions are hypothesized to play a key role in many aspects of vascular function. In an effort to evaluate the importance of connexins in vascular function we took advantage of the fact that a Cx43-LacZ fusion protein has been reported to effectively reduce dye transfer in NIH 3T3 fibroblasts by acting as a dominant negative construct. We explored the use of this dominant negative construct in cultured vascular smooth muscle (VSM) cells and in transgenic mice. We examined the viability of cultured VSM cells expressing the Cx43-LacZ fusion protein under the control of a cytomegalovirus promoter. We also selectively expressed the dominant negative construct in the endothelial cells of transgenic mice under the control of a Tie 2 promoter. Transient transfection of cultured VSM cells led to good initial expression of the Cx43-LacZ fusion protein as evidenced by X-gal staining. Following 10 days of G418 selection, 300 cell clones were examined. None expressed the fusion protein, based on X-gal staining and Western blot analysis, but all contained the transgene, based on PCR analysis. The fusion protein was expressed in a few isolated cells, suggesting that cell division was inhibited by the fusion protein. In agreement with this finding was the fact that expression of the Cx43 LacZ fusion protein was not observed in any of seven Tie 2-Cx43-LacZ transgenic mouse lines. Moreover, a very low yield of mice carrying the transgene was observed (7/136; 5.1%). Analysis of 65 embryos at embryonic day 11.5 showed similar results. These data strongly suggest that the expression of the Cx43-LacZ fusion protein prevents the formation of both stable clones and transgenic animals. This may be due to a cytotoxic effect of the dominant negative construct or to the fact that successful cell propagation is not possible if gap junctional transmission is completely blocked. PMID- 11399893 TI - Alterations in reactivity of small arterioles in rat skeletal muscle as a result of chronic ischaemia. AB - In a model of chronic hind limb ischaemia, we examined whether impaired muscle blood flow, particularly during exercise, is partly due to modification of the reactivity of skeletal muscle resistance vessels by prolonged low blood flow. Two or 5 weeks after unilateral iliac artery ligation, terminal (A4) and preterminal (A3) arterioles of extensor digitorum longus muscle were viewed by intravital microscopy using epi-illumination, and diameter changes to topical application of endothelium-dependent (bradykinin, acetylcholine) and endothelium-independent (adenosine, sodium nitroprusside and noradrenaline) agonists measured. Chronic ischaemia had no effect on resting diameters of A3 or A4 vessels. Two weeks after ligation, dilation to bradykinin was attenuated by 75% for A3 and 50% for A4 arterioles (p < 0.01 vs. control) and responses to acetylcholine were reversed from dilation to constriction (A3: control diameter change +29%, 2-week-ligated 17%; A4: control 18%, 2-week-ligated -13%). Five weeks after ligation, these effects were still apparent and, additionally, dilation to adenosine and sodium nitroprusside and constriction to noradrenaline were reduced. Thus, impaired dilation, most likely due to endothelial dysfunction, is an early manifestation of altered reactivity in the microcirculation of chronically ischaemic muscles, with functional impairment of vascular smooth muscle as a later consequence. These changes occurred despite modest improvements in muscle blood flow and perfusion pressure over the same time. These changes will act to the detriment of blood flow in contracting muscles and could limit the outcome of interventions to restore flow such as angioplasty or surgical bypass. PMID- 11399894 TI - Nonvasomotor influence of sodium nitroprusside on arteriolar remote response to methacholine. AB - Vascular communication functions to facilitate blood distribution within tissues and can be demonstrated as conducted vasomotor responses. This study was designed to determine if local application of sodium nitroprusside (SNP) would affect arteriolar function at remote sites. In the cheek pouch of anesthetized hamsters, local application of SNP and nifedipine caused arteriolar dilation only at the site of application (7.1 +/- 0.5 and 7.4 +/- 0.6 microm), but not at remote sites. The application of SNP enhanced subsequent remote, nitric oxide (NO) independent dilation in response to methacholine, which was applied at a site upstream from the SNP application site (6.7 +/- 0.7 versus 4.5 +/- 0.7 microm for methacholine alone). This potentiating effect was also observed following application of 3-morpholinosydnonimine, but not following nifedipine. This nonvasomotor influence of SNP was not affected by N(omega)-nitro-L-arginine (L NA), tetrodotoxin (TTX), Gap 27 peptide or halothane. Attenuated local dilation in response to methacholine by L-NA could be partially recovered following downstream application of SNP, suggesting that SNP-induced potentiation was associated with enhanced vasodilatory signals at the methacholine application site. Thus, our results suggest that SNP induces nonvasomotor signals in arterioles to affect the network distribution of blood flow. Intrinsic NO, TTX sensitive Na(+) channels and gap junctional communication do not seem to play a major role in the conduction of the nonvasomotor signals. PMID- 11399895 TI - Life and death cell labeling in the microcirculation of the spontaneously hypertensive rat. AB - Spontaneously hypertensive rats (SHRs) have elevated numbers of apoptotic cells. However, the extent and pattern of cell death at the microvascular level is unexplored. We developed a technique to determine early forms of cell death in vivo in the mesentery by use of the life/death indicator ethidium bromide (EB). The mesenteric microvasculature was superfused with 5 microM EB for a period of 3 min, rinsed and immediately viewed by digital fluorescence microscopy. EB positive cell structures were observed both in the wall of microvessels as well as in the tissue parenchyma. The microvessels had about 2--4 EB-positive cell structures per 100 microm of vessel length. Larger arterioles (>25 microm) in the SHR had an increased EB-positive structure density. After normalization of the blood pressure in the SHR with adrenalectomy, no significant differences remained between Wistar-Kyoto (WKY) rats and SHRs. After dexamethasone treatment, the adrenalectomized SHRs had a higher EB-positive cell density in the smaller class of microvessels than the WKY rats. In addition, EB-positive cell fragments (0.5- 2 microm) were observed in the mesentery microvessel wall, and with TUNEL labeling, they were demonstrated to represent DNA fragments. The percentage of microvessels with EB-positive fragments was higher in the SHR arterioles and capillaries. Capillaries and larger venules (>30 microm) in the SHR had higher levels of cell fragments per vessel length. After adrenalectomy, no significant differences remained between WKY rats and SHRs in any of the microvessel categories. When adrenalectomized rats were treated with dexamethasone, a higher number of EB-positive fragments was detected in the wall of SHR capillaries. These results indicate that the mesentery microcirculation in both strains is subject to an early and nonuniform pattern of cell death, as detected by EB, but is enhanced in selected individual microvascular segments of the SHR by a glucocorticoid-driven mechanism. PMID- 11399896 TI - Assessing the homogeneity of the elastic properties and composition of the pig aortic media. AB - Most previous studies of arterial wall elasticity and rheology have assumed that the properties of the wall are uniform across the thickness of the media and, therefore, that the relationship between stress and strain may be described by a constitutive equation based on a single strain energy function. The few studies where this assumption has been questioned, focussed on differences between the adventitia and the media rather than on differences within the media itself. Here, we report in vitro elasticity and residual strain measurements performed separately on the inner and outer half of the pig aortic media, together with a histomorphometric assessment of the radial distribution of elastin, collagen and smooth muscle cell numbers. Although we found that the pressure-diameter relationships of the two halves were dissimilar, when allowance was made for their different unloaded dimensions, their material properties agreed closely, a result in keeping with the observed uniform radial distribution of scleroprotein and vascular smooth muscle. We also found a difference in the opening angle (which is often taken as a measure of residual strain) between the inner and outer medial halves. However, strain analysis showed that the opening angle is an extremely sensitive measure of residual strain and that the difference in the actual magnitudes of residual strain between the two halves of the media was small. We conclude that the media of the porcine thoracic aorta has similar elastic properties throughout its thickness and that this uniformity is matched by a uniform distribution of matrix protein and vascular smooth muscle cells. Furthermore, the distribution of strain in the media can adequately be described by a single-layer model with uniform elastic properties throughout its thickness. PMID- 11399897 TI - Epoxygenase metabolites contribute to nitric oxide-independent afferent arteriolar vasodilation in response to bradykinin. AB - In the kidney, epoxyeicosatrienoic acids (EETs) have been suggested to be endothelium-derived hyperpolarizing factors (EDHFs). The aim of the present study was to determine the contribution of EETs to the preglomerular vasodilation elicited by bradykinin. Sprague-Dawley rats were studied utilizing an in vitro perfused juxtamedullary nephron preparation. The afferent arteriolar diameter was determined and the diameter averaged 19 +/- 1 microm (n = 26) at a renal perfusion pressure of 100 mm Hg. Addition of 1, 10 and 100 nM bradykinin to the perfusate dose-dependently increased afferent arteriolar diameter by 5 +/- 1, 12 +/- 2 and 17 +/- 2%, respectively. The nitric oxide inhibitor N(omega)-nitro-L arginine reduced bradykinin-induced afferent arteriolar vasodilation by 50%, and the diameter increased by 9 +/- 2% in response to 100 nM bradykinin. Epoxygenase inhibitors N-methylsulphonyl-6-(2-propargyloxyphenyl)hexanamide or miconazole greatly attenuated the nitric oxide-independent component of the vasodilation elicited by bradykinin. Cyclooxygenase (COX) inhibition attenuated the nitric oxide-independent vasodilation elicited by 1 nM bradykinin but did not significantly affect the vascular response to 100 nM bradykinin. Combined inhibition of nitric oxide, COX and epoxygenase pathways completely abolished bradykinin-mediated afferent arteriolar vasodilation. In additional studies, renal microvessels were isolated and incubated with bradykinin and samples were analyzed by NICI/GC/MS. Under control conditions, renal microvascular EET levels averaged 49 +/- 9 pg/mg/20 min (n = 7). In the presence of bradykinin, EET levels were significantly higher and averaged 81 +/- 11 pg/mg/20 min (n = 7). These data support the concept that EETs are EDHFs and contribute to the nitric oxide independent afferent arteriolar vasodilation elicited by bradykinin. PMID- 11399898 TI - A common phenotype associated with atherogenesis in diverse mouse models of vascular lipid lesions. AB - The introduction of a range of different genetic modifications in mice results in altered lipoprotein metabolism and the development of vascular lipid lesions. At present, however, it is unclear to what extent the molecular events underlying lipid lesion formation are similar in these different mouse models of atherosclerosis. The aim of this study was to compare the protein expression pattern of lipid lesions from seven different mouse lines with varying susceptibility to vascular lipid lesion development, to determine to what extent lesions induced by different genetic interventions have a similar composition. The proteins we have measured, using quantitative immunofluorescence, are proteins whose expression is known to be modulated during atherogenesis in humans, including plasminogen activator inhibitor (PAI)-1, transforming growth factor (TGF)-beta 1, osteopontin and the macrophage marker CD11b. In all the mice lines we have investigated, PAI-1 was elevated wherever lesions developed. Active TGF-beta was depressed in the vessel wall of mice which developed lipid lesions, particularly in the intima. In contrast, TGF-beta 1 antigen (active plus latent TGF-beta 1) was increased at lesion sites. Accumulation of osteopontin and, with the marked exception of apolipoprotein(a) transgenic mice, tissue macrophages occurred at sites of lipid deposition in the vessel wall. Each lesion, irrespective of its size and the mouse strain in which it developed, had similar amounts of PAI-1, active TGF-beta and osteopontin per unit area of lesion. These data are consistent with a common phenotype accompanying atherogenesis, irrespective of the genetic basis of susceptibility. PMID- 11399899 TI - Failure of L-nitroarginine to inhibit the activity of aortic inducible nitric oxide synthase. AB - Nitric oxide (NO) is produced by a family of three isoenzymes: the endothelial, inducible and neuronal NO synthases. L-Nitroarginine methyl ester (L-NAME) is the most commonly used inhibitor of NO synthase activity. The goal of the present study was to evaluate to what extent L-nitroarginine (L-NA), the in vivo circulating metabolite of L-NAME, blocks NO production in the rat aorta depending on the NO synthase isoform expressed (and evidenced by Western blotting) and on the presence or absence of the extracellular NO synthase substrate L-arginine (100 microM, i.e. the plasma concentration). Intact [endothelium present (E+)] control aortic rings express mainly endothelial NO synthase. L-NA (30--100 microM) induced a dose-dependent contraction (due to blockade of the relaxant properties of NO) irrespective of the presence or absence of L-arginine. In deendothelialized (E-) control aortic rings, the three isoforms of NO synthase are virtually absent (as demonstrated by Western blotting) and L-NA does not elicit any contractile effect. E- aortic rings from lipopolysaccharide (LPS) treated rats express mainly inducible NO synthase. In these rings, L-NA induced a dose-dependent (0--100 microM) contraction in the absence of extracellular L arginine, whereas L-arginine (100 microM) completely abrogated the contractile effect of the NO synthase inhibitor. Chronic L-NAME administration (50 mg/kg/day for 4 weeks) elicited the aortic expression of inducible NO synthase, but to a lesser extent (about 5-fold) than in LPS-treated rat aorta. The average plasma concentration of L-NA was 50 +/- 10 microM in these rats. In E- rings from these L-NAME-treated rats, L-NA induced a similar contractile response (but smaller in magnitude) to that observed in LPS-treated rat aorta. Altogether, these data suggest that (1) in the presence of a physiological concentration of extracellular L-arginine, L-NA fails to inhibit inducible NO synthase, and (2) chronic L-NAME administration, at a dose commonly given to block NO production in vivo, leaves the activity of inducible NO synthase unaffected. PMID- 11399900 TI - Oleate oxidation and mitochondrial substrate selection in vascular smooth muscle. AB - The purpose of this study was to determine the contribution of long-chain fatty acids relative to other mitochondrial substrates in active vascular smooth muscle (VSM). Hog carotid arteries were isometrically contracted in physiological saline solution containing 0.71 mM U-(13)C-oleic acid (bound to albumin at a ratio of 6.8:1), 5 mM 1-(13)C-glucose and 1 mM acetate in the presence or absence of 5 mM carnitine for 6 h at 37 degrees C. Substrate oxidation was determined using (13)C isotopomer analysis of glutamate. Although oxidation of oleic acid could not be measured at physiological concentrations [0.5 mM (1:1)], oleic acid oxidation was approximately 5% of the total substrates oxidized at the higher concentration examined. Although insignificant, carnitine increased oleic acid oxidation to approximately 8%, and resulted in a decrease in endogenous lipid oxidation, which was 2-12% of the total substrates oxidized. Oxidation of glucose and acetate did not significantly change due to the inclusion of oleic acid in the incubation solutions. Therefore, we conclude that exogenous long-chain fatty acids are minor contributors to substrate oxidation (approximately 5%) in VSM compared to other mitochondrial substrates, such as glucose and acetate, which account for approximately 80% of the substrates oxidized by VSM. PMID- 11399901 TI - The time course of tumor necrosis factor-alpha, inducible nitric oxide synthase and vascular endothelial growth factor expression in an experimental model of chronic myocardial infarction in rats. AB - An injury to the heart due to myocardial infarction may progress to heart failure. Among the cytokines and growth factors whose interactions promote remodeling of the heart, increased expression of tumor necrosis factor (TNF) alpha, inducible nitric oxide synthase (iNOS) and vascular endothelial growth factor (VEGF) has been found. However, little is known about the sequence of gene expression during the progression of heart injury. In the present study, male Sprague-Dawley rats were used for experimental myocardial infarction performed by ligation of the left anterior descending coronary artery. TNF-alpha, iNOS and VEGF expression was assessed by reverse transcription polymerase chain reaction. Localization of TNF-alpha, VEGF and iNOS protein was assessed by immunohistochemistry. An in vitro proliferation (BrdU incorporation) and differentiation (tube formation) assay of human umbilical vein endothelial cells was performed. The expression of TNF-alpha, iNOS, VEGF(164) and VEGF(188) was observed during the whole period after myocardial infarction (on days 1, 4, 11, 28 and 40), whereas VEGF(120) was found only on day 1 and 4. The most intense immunostaining for TNF-alpha was observed at the border zone. The iNOS immunostaining was initially located in the endothelium, whereas later it was also present in the walls of larger vessels. The VEGF protein was present in the border zone. No gene expression or immunostaining was detected in sham-operated rats. The in vitro experiments showed both proangiogenic (low TNF-alpha concentration, short period of incubation) and antiangiogenic (high TNF-alpha concentration, long period of incubation) effects of TNF-alpha. The expression of TNF-alpha and iNOS genes with the concomitant occurrence of a decrease in VEGF(120), VEGF(188) and VEGF(164) protein could be related to insufficient angiogenesis and may suggest the possible involvement of these events in remodeling after myocardial infarction. PMID- 11399902 TI - Expression of corticotropin-releasing hormone type 1 receptor in paraventricular nucleus after acute stress. AB - We have previously proposed the existence of ultrashort loop-positive feedback regulation of corticotropin-releasing hormone (CRH) in the hypothalamus. To gain a better understanding of this effect, we performed double-label in situ hybridization to identify the neurons in the paraventricular nucleus (PVN) that express CRH type 1 receptor (CRH-R1) following stress. We also conducted immunohistochemistry to determine whether CRH-R1 mRNA was translated to CRH-R1 protein in the PVN. Thirty-minute restraint stress given to male Wistar rats increased c-fos mRNA expression primarily in the CRH-producing neurons of the parvocellular PVN. Small numbers of vasopressin and oxytoxin-producing cells were also labeled by c-fos probes. Approximately 70% of CRH-R1 positive neurons exhibited CRH mRNA 2 h after the beginning of stress, while only a small percentage of the vasopressin and oxytocin-producing cells coexpressed CRH-R1 mRNA. CRH-R1 immunoreactivity, which was detected in the perikarya and fibers of PVN neurons, appeared to increase in response to stress, though this was not statistically significant. Pretreatment with a selective CRH-R1 antagonist, CP 154,526, significantly attenuated stress-induced corticotropin (ACTH) secretion as well as c-fos mRNA expression in the PVN. These results demonstrate that acute stress increases neuronal activation and CRH-R1 mRNA expression primarily in CRH producing neurons of the parvocellular PVN, that CRH-R1 message is translated to CRH-R1 protein, and that PVN neurons are activated at least in part through CRH R1 under acute stress. The data further support the possibility of feedback regulation of CRH itself in CRH-producing neurons. PMID- 11399903 TI - A short period of maternal nutrient restriction in late gestation modifies pituitary-adrenal function in adult guinea pig offspring. AB - Altered fetal environment can program the hypophyseal-pituitary-adrenal (HPA) axis development and thus affect endocrine function in later life. We hypothesized that 48 h of maternal nutrient restriction during the period of maximal fetal brain growth alters HPA function in adult offspring and leads to modified blood pressure regulation. Pregnant guinea pigs (n = 15) were deprived of food (water ad libitum) or fed normally (n = 13) on days 50 and 51 of gestation, after which they were all fed normally (birth = 68 days). Carotid artery and jugular vein catheters were implanted in adult guinea pig offspring (day 65). Animals were treated with corticotropin (ACTH(1-24); 0.5 microg/kg), corticotropin-releasing hormone (CRH; 0.5 microg/kg) and insulin (5 units/kg), and pituitary-adrenal responses were measured. Guinea pigs were then euthanized and pituitaries removed for analysis of pro-opiomelanocortin (POMC) and glucocorticoid receptor (GR) mRNA levels. There was no effect of prenatal treatment on body weight, blood pressure or heart rate. In male offspring, both basal ACTH (p < 0.007) and basal cortisol (p < 0.05) levels were significantly reduced in animals whose mothers had been nutrient restricted (NR). In contrast, in female offspring, basal plasma ACTH was not different between offspring from NR mothers and controls; however, basal plasma cortisol concentrations were significantly (p < 0.01) elevated at 13.00 h in females born to NR mothers. Responses to HPA challenge were different between offspring from NR mothers and control offspring, and these differences were consistent with alterations in basal adrenocortical function. There was no effect of prenatal treatment on POMC mRNA levels in the pars distalis or pars intermedia. However, GR mRNA levels were significantly (p < 0.05) reduced in adult female offspring born to NR mothers. In conclusion, 48 h of maternal nutrient restriction during pregnancy has a long term effect on HPA function in adult offspring, and this effect is highly sex specific, but does not result in alteration of blood pressure. PMID- 11399904 TI - Hypothalamo-pituitary-adrenal axis activity is related to the level of central arousal: effect of sleep deprivation on the association of high-frequency waking electroencephalogram with cortisol release. AB - The temporal and quantitative interrelationships between the hypothalamo pituitary-adrenal (HPA) axis activity and the level of central arousal were studied in 10 healthy young men during daytime wakefulness. Two experimental sessions were conducted randomly between 09.00 and 18.00 h, once after nocturnal sleep and once after a night of total sleep deprivation. Spectral analysis of serial waking electroencephalography (EEG) from a short target fixation task repeated every 10 min was undertaken, along with an estimation of cortisol secretory profiles by deconvolution of plasma radioimmunoassay measures obtained from continuous blood withdrawal with regular sampling at a 10-min interval. Following nocturnal sleep, a temporal association between the HPA axis activity and the waking EEG activity was found, cortisol secretory rate following changes in frontal gamma (20-45 Hz) band power by 10 min (average R = 0.458, p < 0.001). Although it remained significant (average R = 0.276, p < 0.05), the association strength decreased significantly following total sleep deprivation (p < 0.05, Wilcoxon test). Cortisol plasma level, secretory rate and pulse amplitude were increased as well as waking EEG power in the delta (0.5-5.5 Hz), theta (5.5-8.5 Hz) and gamma frequency bands (all p values <0.05, Student t tests). The sleep deprivation-related increases in cortisol secretory rate and waking EEG gamma activity were quantitatively associated (R = 0.504, p < 0.05). These results support the existence of a common ultradian regulatory mechanism, co-ordinating HPA axis activity to the level of central arousal in man, which seems involved in the sleep deprivation-induced hyper-arousal. PMID- 11399905 TI - Differential effect of serotonin 5-HT(1A) receptor antagonists on the secretion of corticotropin and prolactin. AB - Serotonin (5-HT) participates in the neuroendocrine regulation of corticotropin (ACTH) and prolactin (PRL) secretion. We investigated the involvement of the 5 HT(1A) receptor in the mediation of ACTH and PRL response to the 5-HT(1A) receptor agonists 8-OH-DPAT and 5-HT, the 5-HT precursor 5-hydroxytryptophan (5 HTP) and restraint stress in male rats. Prior intracerebroventricular (i.c.v.) infusion of the 5-HT(1A) antagonists WAY-100635 and LY-206130 inhibited PRL and ACTH responses to i.c.v. infusion of 8-OH-DPAT, 5-HT as well as to 5-HTP administered systemically in combination with the 5-HT reuptake inhibitor fluoxetine (Flx). Infused i.c.v. NAN-190 inhibited the ACTH response to 8-OH-DPAT i.c.v. Intraperitoneal (i.p.) pretreatment with WAY-100635 inhibited ACTH and PRL responses to 8-OH-DPAT, whereas LY-206130 only inhibited the PRL response and NAN 190 had no effect. Injected i.p., the antagonists had no effect on 5-HT-induced hormone secretion, whereas the ACTH-stimulating effect of 5-HTP + Flx was increased by WAY and NAN. A 5-min restraint stress increased ACTH and PRL secretion; the ACTH, but not the PRL response to stress was inhibited by prior administration of WAY-100635 or LY-206130 either i.c.v. or i.p. NAN-190 had no effect on any stress response tested. It is concluded that (1) the three 5-HT(1A) antagonists used in our study have differential effects on stimulated hormone responses, (2) the antagonists exert different effects when administered systemically or centrally, and (3) the 5-HT(1A) receptor is involved in restraint stress-induced ACTH, but not PRL secretion. PMID- 11399906 TI - GABA(B) receptors in anterior pituitary cells. Mechanism of action coupled to endocrine effects. AB - The activation of pituitary GABA(B) receptors by the specific agonist baclofen inhibits pituitary hormone secretion in vitro. Here we studied the mechanism of action of GABA(B) receptors in rat adenohypophysis. Anterior pituitary cells were obtained by trypsinization and were either plated for hormonal studies and cAMP determination or incubated in FURA 2AM for calcium measurements. Baclofen (BACL: 1 x 10(-5) M) significantly inhibited basal and thyrotropic releasing hormone (TRH)-stimulated (1 x 10(-7) M) PRL secretion in anterior pituitary cells from proestrous rats. In the presence of pertussis toxin (PTX: 150 ng/ml, 20 h), which leads to the uncoupling of the G(i/o)-protein from the receptor, both effects of BACL were abolished while the effect of dopamine (DA: 1 x 10(-8) M), used as an inhibitory control, was reduced from 70 to 25%. PTX also reversed BACL-induced inhibition of gonadotropin-releasing hormone (GnRH)-elicited luteinizing hormone (LH) secretion in anterior pituitary cells from 15-day-old female rats. In addition, though working in a pituitary mixed cell population, in which only some cell types possess GABA(B) receptors, BACL (1 x 10(-5) M) attenuated the forskolin-induced (0.5 microM) increase in cAMP. This effect was prevented by co incubation with the antagonist 2 hydroxysaclofen and by preincubation with PTX. BACL (5 x 10(-5) M) and DA (5 x 10(-7) M) inhibited basal intracellular calcium concentrations ([Ca(2+)](i)) in pituitary cells and the effect of the latter was significantly stronger. The effect of BACL on [Ca(2+)](i) was abolished after preincubation with PTX. In the presence of the potassium channel blocking agents barium (200 microM and 1 mM) and tetraethylammonium (10 mM), BACL was still able to inhibit [Ca(2+)](i). Blockade of voltage-sensitive calcium channels (VSCC) with either verapamil (5 x 10(-6) M) or nifedipine (1 x 10(-6) M) completely abolished the effect of BACL on [Ca(2+)](i). In the presence of 12.5 mM potassium concentration baclofen significantly inhibited [Ca(2+)](i). In conclusion, our results describe the negative coupling of adenohypophyseal GABA(B) receptors to VSCC through PTX-sensitive G-proteins. These characteristics suggest a resemblance of these receptors to the typical presynaptic GABA(B) sites described in the central nervous system. PMID- 11399907 TI - Differential inhibition of growth hormone secretion by analogs selective for somatostatin receptor subtypes 2 and 5 in human growth-hormone-secreting adenoma cells in vitro. AB - Somatostatin (SRIH), a cyclic tetradecapeptide hormone originally isolated from mammalian hypothalamus, is a potent suppressor of pituitary growth hormone (GH) secretion. SRIH acts through a family of G-protein-coupled membrane receptors containing seven transmembrane domains. Five genes encoding distinct SRIH receptor (SSTR) subtypes have so far been cloned in human and other species and termed SSTR1-5. In human somatotrophe pituitary adenomas GH secretion is controlled by both SSTR2 and SSTR5. However, in clinical practice only somatostatin analogs selective for SSTR2 (octreotide and lanreotide) are available. This may explain why clinical and in vitro responses to these analogs in acromegaly are only partial. In this study, we investigated the inhibitory effect of two new SRIH analogs with high selectivity for SSTR2 (NC-4-28B) and SSTR5 (BIM-23268) and compared it to that of native somatostatin (SRIH-14) on a large number of GH-secreting adenomas obtained by transphenoidal neurosurgery. Tissues from 16 adenomas were enzymatically dispersed and plated in 24-well dishes at 50,000 cells/well. After 3 days, groups of three wells were incubated for 4 h with medium alone, SRIH-14 or analogs NC-4-28B or BIM-23268, at the concentrations of 0.01, 0.1 and 1 microM. Our results show that 9 out of 16 adenomas were responsive (GH suppression: 20-40% vs. control, p < 0.05) to SRIH. In this group only 4 adenomas showed similar responses to both selective analogs, with 2 nonresponders (expression of other SRIH receptor subtypes) and 2 responders (concomitant expression of SSTR2 and SSTR5) to both analogs. GH release was selectively inhibited by NC-4-28B in 3 adenomas and by BIM-23268 in the remaining 2 adenomas, suggesting predominant expression of SSTR2 and SSTR5, respectively. SRIH failed to inhibit GH release in 7 adenomas (43%). Interestingly, in that group a better inhibitory effect was obtained with BIM 23268 (5 out of 7 adenomas) than with NC-4-28B, suggesting expression of a few SSTR5 receptors only, or of both SSTR2 and SSTR5, respectively. We conclude that the availability of somatostatin analogs selective for SSTR5 will enhance the treatment potency and spectrum in acromegaly. PMID- 11399908 TI - Distribution and stimulation by gastrin-releasing peptide of protein kinase C subfamilies in insulin-secreting cells. AB - The role of the different isoforms of protein kinase C (PKC) in modulating insulin secretion is still widely unknown. The aim of our studies was to investigate which isoforms are influenced by gastrin-releasing peptide (GRP), a neuropeptide which has been shown to modulate insulin secretion by activating PKC. Presence of PKC isoforms alpha, beta, gamma, delta, epsilon and zeta was tested by immunoblot analysis in whole pancreatic islets of mouse and rat and in the insulinoma cell line RINm5F. Effects of GRP, the truncated peptide GRP1-16 and KCl were also measured on translocation of PKC isoforms. In pancreatic islets of mouse and rat, the PKC isoforms alpha, beta, gamma, delta, epsilon and zeta could be detected. No PKCgamma activity was present in the pancreatic tumor cell line RINm5F. Incubation of mouse or rat islets or of RINm5F cells with GRP induced translocation of the PKC isoforms alpha, beta and zeta. The N-terminal portion of the peptide GRP1-16 induced partial translocation only of the PKC isoforms alpha, beta and zeta in mouse and rat islets in 4 out of 10 cases, but failed to show any effect on PKC isoforms in RINm5F cells. Depolarization of the islets by KCl did not translocate any tested PKC isoform. However, incubation with GRP followed by depolarization with KCl led to translocation of the PKC isoforms alpha, beta and zeta. It is suggested that PKC alpha, beta and/or zeta may play a role in the modulation of insulin secretion by GRP. PMID- 11399909 TI - Both substance P and its receptor are expressed in mouse intestinal T lymphocytes. AB - Substance P (SP), one of the most prevalent neuropeptides in gut, has been reported to have potent immune modulatory effects as a proinflammatory agent. The synthesis of SP and SP receptor expression in intraepithelial and lamina propria T lymphocytes of mouse intestine was investigated. Using RT-PCR analysis, it was demonstrated that SP receptor mRNA was exclusively expressed in intraepithelial and lamina propria T lymphocytes as well as their purified CD4+, CD8+ and CD4-CD8 CD3+ subsets. Messenger RNAs (mRNAs) for the two precursors of SP, beta and gamma preprotachykinin-A, were also detected. These results were consistent in lymphocytes from both epithelium and lamina propria of small and large intestines, although the frequencies and/or intensities of mRNA expression varied. However, none of the findings could be repeated in splenic T lymphocytes. Activation of splenocytes with anti-CD3epsilon-chain mAb and PMA did not induce expression of SP or its receptor mRNAs. Furthermore, both cytoplasmic and surface bound SP was demonstrated in intestinal T lymphocytes using dual color immunocytochemistry and immunoflow cytometry. In vitro treatment with SP did not significantly change the size of the SP-immunoreactive T cell population, indicating the presence of SP receptor on intestinal T lymphocytes as well as in vivo binding of endogenously released SP. Our data suggest that SP production and SP receptor expression are distinctive for mouse intestinal mucosal immunity and that SP may act as a modulator of an ongoing controlled inflammation in normal gut, by acting through its specific receptor on T lymphocytes in an autocrine and/or paracrine pattern. PMID- 11399910 TI - Causes and consequences of adynamic bone disease. PMID- 11399911 TI - Molecular genetics in IgA nephropathy. AB - Evidence from both genotypic and phenotypic perspectives is considered that patients may be genetically predisposed to IgA nephropathy (IgAN) or Henoch Schonlein purpura (HSP) or that a factor(s) might exclusively contribute to their progression to chronic renal failure. In contrast to most other renal diseases, both IgAN and HSP are uncommon in blacks; this is unexplained but is not due to their low frequency of the A2m(1) allotype. The association of these diseases or their progression with a variety of abnormalities of IgA immunobiology in patients and their families has not been linked to any genotype; similarly, no HLA antigen has been positively or negatively associated in any consistent way. Although complement factor 3 universally accompanies IgA glomerular deposition, complement pathway abnormalities are only sporadically reported with either IgA deposition or disease progression. Whether angiotensin-related polymorphism including the converting enzyme alleles have a specific predictable role, particularly in the progression of renal failure in IgAN, remains problematic. The promising possibility that a structural defect in IgA1 due to an as yet unidentified genetic defect accounts for the deposition of IgA is considered in some detail. Nevertheless, the genetic mechanism(s) of progressive renal failure, whether exclusive to IgAN or to glomerular diseases generally, is of paramount importance. PMID- 11399912 TI - Relationship between fluctuation pattern of blood pressure during hemodialysis treatment and cardiovascular morphology: an autopsy study of 53 cases. AB - BACKGROUND: Cardiovascular morphological changes are often conspicuous in autopsy examination of chronic hemodialysis (HD) patients. On the other hand, the fluctuation pattern in blood pressure (BP) during HD treatment varies from one patient to another. Cardiovascular changes may correlate with clinical findings including BP fluctuation patterns during HD, although no autopsy studies have previously examined this issue. METHODS: In this study, 53 autopsies of patients who had been on chronic HD were reviewed. We determined the relationship between BP fluctuation during HD treatment along with stable and cardiovascular morphology, including heart weight, ventricular wall thickness, circumferences of the valves and the severity of aortic arteriosclerosis and coronary stenosis. Patients were divided into 4 groups according to the pattern of BP fluctuation during HD treatment at about 6 months before death: group 1 (n = 13), symptomatic hypotension and/or decline pattern during HD; group 2 (n = 11), continuously high BP during HD treatment; group 3 (n = 17), continuous normal BP during HD treatment, and group 4 (n = 12), continuously low BP without symptomatic hypotension during HD treatment. RESULTS: Heart weight and ventricular wall thickness were greatest in group 2. The scores for aortic arteriosclerosis in groups 1 and 2 were higher than in groups 3 and 4. The coronary stenosis index was significantly higher in group 1 than in the other groups, and that in group 2 was higher than in group 4. Multiple regression analysis showed that age, HD duration and pulse pressure were independent variables for the score of arteriosclerosis, and the decline pattern of BP fluctuation during HD and pulse pressure were independent variables for coronary stenosis index. CONCLUSIONS: Our results suggest that certain clinical parameters including BP during HD may reflect cardiovascular morphological changes in stable HD patients, although further examination, such as 24-hour blood pressure measurement is recommended to elucidate the pathophysiology of cardiovascular diseases in HD patients. PMID- 11399913 TI - Virological and histological responses to one year alpha-interferon-2a in hemodialyzed patients with chronic hepatitis C. AB - BACKGROUND: alpha-Interferon-2a (IFNalpha) alone is a therapy of limited proven benefit for non-uremic patients with chronic hepatitis C virus (HCV) infection. In dialyzed patients, such an effect is suggested on small short-term studies without sufficient clinical and virologic follow-up to document any sustained effect. PROTOCOL: Twelve chronically hemodialyzed patients with chronic hepatitis C and waiting for renal transplantation were included in a prospective open study of treatment with IFNalpha. We used, as did others, doses of 3 million units (MU), three times a week, but for a longer period of treatment of 12 months. Follow-up was continued for 6 months after the end of IFNalpha in order to document any sustained biochemical, virological and histological responses. RESULTS: Aminotransferase levels returned to the normal range within 1-2 months of treatment in all patients in whom they had been elevated at baseline. At 1 month of treatment, serum HCV-RNA was not detected in 5 (41%) patients and in 9 (75%) at 12 months. A sustained virological response was documented in 4 (33%) patients 6 months after the end of treatment. Relapse occurred in 5 patients within 2 months after IFNalpha withdrawal. HCV genotype was not predictive of any sustained response. At inclusion, using the histologic Metavir scoring system, half of the patients had low-grade cytolytic activity and none had cirrhosis. After IFNalpha, liver biopsy specimens were available from 9 patients and showed histologic improvement in 3. IFNalpha tolerance was poor, inducing a 5% mean weight loss and the acute rejection of two nonfunctioning kidney grafts. CONCLUSION: This study documents that administration of IFNalpha at 3 MU three times a week, for 12 months, in hemodialysis patients with chronic hepatitis C was efficient for clearing the serum of HCV-RNA in 75% of the patients. A sustained response was maintained in one third of these patients after cessation of IFNalpha, and was predicted by the early serum clearance of the virus within the first 2 months of treatment. We confirm that a 12-month treatment period carries a higher sustained response rate than shorter treatment periods. These encouraging results call for larger studies in uremic patients, using IFNalpha alone or in association with new antiviral drugs. PMID- 11399914 TI - Evidence for alterations in circulating low-molecular-weight antioxidants and increased lipid peroxidation in smokers on hemodialysis. AB - BACKGROUND/AIM: Cardiovascular disease is the major cause of mortality in dialysis patients, accounting for about 40% of deaths in most large registries. Oxidative stress has been strongly implicated in the pathogenesis of these events. As end-stage renal disease is a state of elevated free radical activity, the aim of the present study was to investigate the negative impact of smoking in 57 male hemodialysis patients. METHODS: The patients, who were 20-85 years of age (mean age 51.0 +/- 14 years), had been on hemodialysis for at least 6 months before participating in this study. Fasting blood sampling for serum lipid, albumin, urate and lipophilic antioxidants such as tocopherols, carotenes, ascorbate and lipid peroxides was performed. RESULTS: The plasma malondialdehyde (MDA) concentration was significantly higher in hemodialysis patients who smoked compared to hemodialysis patients who were nonsmokers (1.92 +/- 0.52 vs. 1.59 +/- 0.42 nmol/ml, p = 0.006). No association was found between levels of MDA in smokers and parameters such as body mass index, serum cholesterol, serum triglycerides and smoking index. There were no significant differences in the plasma levels of uric acid, alpha-tocopherol, gamma-tocopherol, delta-tocopherol, alpha-carotene, beta-carotene and retinol between the two groups. A significantly lower level of plasma ascorbate was observed in hemodialysis patients who smoked compared to the nonsmoking hemodialysis patients or healthy controls (4.59 +/- 4.0 vs. 9.57 +/- 4.0 and 10.16 +/- 4.6 microg/ml, p < 0.05). Moreover, in smokers, the plasma levels of ascorbate were negatively correlated with the levels of plasma MDA (r = -0.43, p < 0.001) of each patient. Partial correlation analysis of the plasma levels of the measured antioxidants and the smoking index revealed a negative correlation between the plasma levels of lipid-normalized lycopene and the smoking index (r = -0.53, p < 0.05). CONCLUSION: Our data suggest that cigarette smoking further increases plasma-circulating products of lipid peroxidation, which are already increased in nonsmoking hemodialysis patients as compared to matched healthy controls. The lower plasma levels of ascorbate in hemodialysis patients who smoke suggest that these patients may be more susceptible to oxidative tissue damage caused by smoking. PMID- 11399915 TI - Appropriate dosing of pilsicainide hydrochloride in patients on hemodialysis. AB - Appropriate dosing of pilsicainide hydrochloride, an anti-arrhythmic drug excreted via the kidney, was investigated in patients on dialysis. Ten chronic hemodialysis patients with coexisting severe palpitation of supraventricular premature contractions (SVPC) were treated with 25 mg of pilsicainide hydrochloride before dialysis. All of their plasma concentrations were maintained within the therapeutic range and their mean dialysis rate was 32%. After 2 weeks, 7 patients were followed with consecutive daily dose treatment. In 3 of them, the dosage was returned to the single pre-dialysis administration because of the elevated plasma concentration reaching the toxic range 1 month after the start of administration. The dose schedule was maintained, and plasma pilsicainide concentrations remained within the therapeutic range during the 6-month follow up. No abnormal findings were found in any parameters of electrocardiography, echocardiography or biochemistry. The number of SVPC diminished > 90% compared to the pretreatment level. PMID- 11399916 TI - Tuberculosis in maintenance dialysis patients. AB - In this investigation, we tried to find the incidence and characteristics of tuberculosis (TB) in dialysis patients previously found only in a small number of cases. We collected the cases of newly diagnosed TB patients in Taiwan during 1997. Simultaneously, all dialysis patients were collected and matched with the TB cases to identify the dialysis patients who had also contracted TB. The annual incidence of the dialysis population was 493.4/100,000, 6.9 times that of the general population (71.1/100,000). The annual incidence for the male dialysis population was 573.3, the incidence was 479.2 for the female dialysis population. The incidence for the general population was 97.1 and 43.7/100,000, respectively. Although the 1-year mortality rate due to TB (1.7 vs. 1.9%, p > 0.05) was similar in both populations, the non-TB mortality was much higher in the dialysis population than that in the general population (25.6 vs. 11.1%, p < 0.05). Finally, the 1-year mortality rate of dialysis patients with TB is 3.3 times higher than that in dialysis patients without TB (27.3 vs. 8.3%, p < 0.05). The findings suggest that uremia modifies the behavior of TB, jeopardizes female and younger dialysis patients, poses a higher risk of extrapulmonary dissemination, and predicts a higher overall mortality. PMID- 11399917 TI - Long-term results of donor-specific blood transfusion with cyclosporine in living related kidney transplantation. AB - Donor-specific blood transfusion (DST) was introduced to achieve better graft survival. However, its benefits are controversial considering the immunosuppression of cyclosporine (CYA) or tacrolimus (Tac), and its long-term effects have not been well discussed. Of the 40 patients who received DST with CYA, 3 (7.5%) became cross-match positive. Of the 37 patients with negative cross match, 34 patients received a one-haplotype-matched kidney and were compared to patients with one-haplotype-matched kidney transplant without preoperative DST (n = 13). Acute rejection within 3 months after transplant was 29.4% in the DST group, and 15.4% in the non-DST group. All rejection episodes were steroid resistant in the non-DST group. If the graft survival rates were calculated excluding non-immunological graft loss, graft survival rate was 91.0 and 72.8% at 5 and 10 years in the DST group, and 83.3% at 5 and 10 years in the non-DST group, respectively. The two graft survival lines converged 7 years and 7 months after transplantation. No beneficial effect of DST was statistically evident under CYA immunosuppression. In terms of the severity of acute rejection or the onset of chronic rejection, DST induced a small benefit, however, which seemed to disappear within 8 years after transplantation. PMID- 11399918 TI - Parathyroidectomy in chronic renal failure: short- and long-term results on parathyroid function, blood pressure and anemia. AB - To evaluate the long-term results of parathyroidectomy (PTX) on parathyroid function, blood pressure and anemia, data of 45 patients with secondary Hyperparathyroidism in dialysis who had undergone PTX were collected retrospectively from 8 different dialysis units. The patients, 25 M and 20 F, mean age 56 +/- 11 years, who were followed up for an average period of 3.3 +/- 2.3 years, were divided into four groups according to the surgical procedure: 19 patients had had a subtotal PTX; 10 patients had undergone total PTX with autotransplantation (AT); 10 patients had had total PTX without AT, and 6 patients had undergone partial PTX. Taking a reduction in intact PTH > 50% as sign of successful PTX, only 5 patients did not attain this result. Considering values of PTH between 20 and 200 pg/ml at the mid-term observation (1-2 years) as the optimal result, values under 20 pg/ml as an expression of permanent hypoparathyroidism, and those above 200 pg/ml as indicating persistent/recurrent hyperparathyroidism, 65.5% of patients operated with subtotal PTX and total PTX + AT had a therapeutic success, versus 31.2% of patients in the other two groups, due to excess permanent hypoparathyroidism and persistent/recurrent hyperparathyroidism; 20 of 45 patients with preoperative hypertension experienced a statistically and clinically significant decrease in blood pressure levels. An increase in serum hemoglobin was also observed, despite a reduction of administered erythropoietin. In conclusion, the results of PTX obtained from this multicenter study are comparable to those reported by single leading centers. Recommended surgical procedures are subtotal PTX and total PTX with AT. The fall in blood pressure in hypertensive patients is clinically significant, and improvement in anemia is also observed with a reduction in erythropoietin dosage. PMID- 11399919 TI - Increased glomerular cell heparan sulfates in vitro by ciclosporin A: a Possible explanation of Its beneficial effect in idiopathic nephrotic syndrome. AB - BACKGROUND/AIM: In idiopathic nephrotic syndrome (INS), ciclosporin A (CsA) was shown to decrease proteinuria, an effect explained by its immunologic and hemodynamic actions. In order to determine whether CsA could have a direct action on glomerular cells, we studied the effect of CsA on glomerular cells in vitro, particularly on glycosaminoglcycans (GAG) and heparan sulfates (HS) which are decreased in INS patients. METHODS: Human glomerular epithelial cells and rat mesangial cells were cultured at various concentrations of CsA. HS were quantified using a cationic membrane after metabolic labeling. RESULTS: Mesangial cell GAG and HS and epithelial cell HS increased significantly when cells were cultured with CsA. For both cell types this increase was prevailing on the secreted fraction of HS in comparison with the cellular fraction. CsA induced also an increase in cellular cAMP levels, but the effect of CsA was not transduced via a cAMP pathway. CONCLUSIONS: CsA is able to increase glomerular GAG and HS in vitro. As this effect of CsA was the opposite effect on glomerular cells to the effect of plasma from INS patients, we conclude that this direct action of CsA on glomerular cells could explain in part the effect of this drug in decreasing proteinuria in INS. PMID- 11399920 TI - Studies on calcium oxalate binding proteins: effect of lipid peroxidation. AB - OBJECTIVE: Urolithiasis and free radicals have long been associated. In this study, we have isolated calcium oxalate monohydrate (COM) binding proteins from rat kidney before and after lipid peroxidation (LPO) and studied its properties on calcium oxalate crystal growth. MATERIALS AND METHODS: LPO was carried out using t-butyl hydroperoxide, cumene hydroperoxide and an ascorbate system. The COM binding proteins from control and peroxidised tissues were isolated using a modified procedure. Protein was extracted using 25 mM EDTA, and the extract was loaded onto a DEAE cellulose column and eluted with Tris-HCl buffer (pH 6.5), 0.05 M NaCl in the above buffer and 0.3 M NaCl in the same buffer. Three major protein fractions were obtained, and they were designated as fractions I, II and III according to their order of elution. The proteins were subjected to calcium oxalate crystal nucleation and aggregation. RESULTS: A positive correlation was observed between LPO and COM adsorption, while a negative correlation was observed between reduced glutathione and COM adsorption. Peroxidised protein did not show any alteration in the elution profile on the DEAE cellulose column. The SH content of the peroxidised fractions were lower than that of the control fractions, but their oxalate binding activities were increased. Peroxidised fraction I promoted crystal growth to a greater extent than the control fraction I. Peroxidised fractions II and III were less inhibitory in nature compared to their control fractions. Light-microscopic examination of the crystals formed in the presence of the peroxidised fractions showed the formation of large aggregates of COM. CONCLUSION: Peroxidation of the renal proteins favoured their adsorption to COM crystals. -SH depletion increased the oxalate binding activity and also their affinity to the COM crystals. The peroxidised fraction I was found to favour the formation of large aggregates, suggesting that peroxidation may be one of the mechanisms altering the crystal inhibitory activity of the proteins in hyperoxaluria. PMID- 11399921 TI - Apolipoprotein(a) phenotype, albumin clearance, and plasma levels of lipoprotein(a) in peritoneal dialysis. AB - This cross-sectional study was undertaken to examine the relationship between plasma lipoprotein(a) [Lp(a)] level and peritoneal dialysis (PD) albumin clearance while controlling for the influence of the apolipoprotein(a) [apo(a)] phenotype. Plasma Lp(a) level, PD albumin clearance, and apo(a) phenotype (high vs. low molecular weight, HMW vs. LMW) were determined in 54 PD patients. Apo(a) phenotypes were 24 LMW and 30 HMW. The plasma Lp(a) level was high (> 65 nmol/l) in 17 of 24 patients with LMW phenotype versus 2 of 30 with HMW phenotype (chi2, p < 0.01). Spearman correlation coefficients of Lp(a) with PD, urine, and total albumin clearances were -0.05 (p = 0.74), -0.04 (p = 0.80), and -0.09 (p = 0.51), respectively. The apo(a) isoform size was the only significant predictor of Lp(a) in multivariate analysis. In this study, there was no association between PD albumin clearance and Lp(a) level. The association between apo(a) phenotype and Lp(a) level is in keeping with studies in the general population. There is a strong genetic influence on Lp(a) level in PD patients. PMID- 11399922 TI - Development of porphyria cutanea tarda in a hemodialysis patient after reactivation of hepatitis C virus infection. AB - Cases of porphyria cutanea tarda (PCT) are occasionally reported in hemodialysis patients. Recently, hepatitis C virus has been recognized as a precipitating factor of PCT. The activity of the liver disease may be critical for the appearance of PCT. In this regard, liver disease reactivation after treatment with interferon alpha (IFN-alpha) is a well-known phenomenon. We report the case of a hemodialysis patient who developed PCT coincidentally with reactivation of liver disease, immediately after treatment with IFN-alpha. Therefore, in the present case, reactivation of hepatitis after IFN-alpha withdrawal could be the triggering factor. The occurrence of a bout of PCT should be considered as a possible complication at the end of IFN-alpha therapy, if reactivation of the disease exists. PMID- 11399923 TI - Membranoproliferative glomerulonephritis associated with hypocomplementemic urticarial vasculitis after complete remission of membranous nephropathy. AB - A 49-year-old-man developed proteinuria in 1978. He was diagnosed as having membranous nephropathy by renal biopsy and was treated with prednisolone. The proteinuria disappeared completely and the treatment was stopped. In 1995, after complete remission, he developed nephrotic syndrome with chronic urticaria and hypocomplementemia. Renal biopsy revealed membranoproliferative glomerulonephritis (type I) and skin biopsy showed leukocytoclastic vasculitis, which was compatible with hypocomplementemic vasculitis syndrome. Steroid therapy was very effective. PMID- 11399924 TI - Evaluation of i-STAT creatinine assay. AB - BACKGROUND: We evaluated a new, 2-min blood creatinine method using the hand-held i-STAT analyzer. Good results have already been reported using this analyzer for 10 methods including electrolytes, TCO2, pH, PCO2, bicarbonate, glucose, hemoglobin and urea for uremic blood, hemodialysate and peritoneal effluent. METHODS: Evaluation included study of imprecision and accuracy. RESULTS: Imprecision studies gave excellent results, including those for reproducibility of 6 solutions with a mean of 10 repeats and coefficients of variation (CVs) of 0.4-3.4%, and also the mean of the differences between 33 duplicate blood specimens which was 2.2% of the specimen mean. To assess accuracy, we compared results of 149 tests by i-STAT and Beckman Synchron CX7 methods. The difference between the two means was 2.6% and the mean of all differences was 10.9% with i STAT results higher, especially when blood creatinine values were < 100 micromol/l (1.1 mg/dl) indicating the need for a slightly higher upper limit of the normal range. The correlation coefficient between the two methods was 0.99, the slope 1.0 and the intercept -5.0 micromol/l (-0.06 mg/dl). We assessed the recommended creatinine correction for variation in PCO2 above and below 40 mm Hg, but our results did not suggest the need for such a correction in our range of 27 64 mm Hg; omission would remove a major method disadvantage. Assays of hemodialysate and peritoneal effluent were also satisfactory. CONCLUSIONS: The i STAT creatinine method is simple and rapid and our evaluation showed satisfactory accuracy and precision. However, results were on average slightly higher than for the Beckman Synchron CX7 method. PMID- 11399925 TI - Antiproteinuric effect of an angiotensin II receptor antagonist in membranous nephropathy. PMID- 11399926 TI - Abdominal aortic aneurysms in hemodialysis patients with autosomal dominant polycystic kidney disease. PMID- 11399927 TI - Ace gene insertion/deletion alleles: transmission from parents to dialysis and nondialysis patients with chronic renal failure. PMID- 11399928 TI - Role of monocyte chemotactic peptide 1 in diabetic nephropathy. PMID- 11399929 TI - Effects of indole on breathing in rats. PMID- 11399930 TI - Early detection, diagnosis and management of choroidal neovascularization in age related macular degeneration: the role of ophthalmologists. AB - Choroidal neovascularization (CNV) secondary to age-related macular degeneration (AMD) is a serious condition which, if unrecognized and untreated, can result in the rapid deterioration of vision. Early detection and prompt referral to a retina specialist may potentially reduce the high risk of severe vision loss. The majority of AMD patients with CNV who present to an ophthalmologist will have been referred by either a primary care physician or an optometrist. Following referral, ophthalmologists will confirm diagnosis and identify the location and composition of the CNV. This information will provide the basis for a decision on what treatment, if any, is indicated. Until last year, laser photocoagulation was the only clinically-proven treatment option for neovascular AMD in large-scale randomized clinical trials, although many patients were not eligible for this treatment. Verteporfin (Visudyne(TM)) therapy is a new treatment option that uses photodynamic therapy in patients with predominantly classic subfoveal CNV secondary to AMD. Retrospective analyses show that the introduction of verteporfin therapy is likely to increase the number of patients with neovascular AMD who can be treated. Early detection, prompt referral and treatment, together with appropriate use of low vision aids, will help to optimize patient outcomes and quality of life. PMID- 11399931 TI - Corneal allograft rejection: current understanding. I. Immunobiology and basic mechanisms. AB - Allograft rejection remains the single largest impediment to success in corneal transplantation. This article briefly reviews our current understanding of some fundamental aspects of corneal immunology and the pathogenetic mechanisms underlying corneal graft rejection. As knowledge increases, it is hoped that a better understanding of the immunobiology may result in improved preventive and therapeutic measures. PMID- 11399932 TI - Transscleral diode laser photocoagulation in refractory glaucoma. AB - BACKGROUND AND OBJECTIVE: Patients with glaucoma refractory to medical or surgical treatment are difficult to treat. The aim of this study was to evaluate the effect of contact transscleral diode laser cyclophotocoagulation (DLPC) on these patients' intra-ocular pressure (IOP) and visual acuity. PATIENTS AND METHODS: 37 eyes of 36 patients who suffered from refractory glaucoma and who had undergone DLPC, were followed at the Eye Clinic of Verona University for an average of 14 +/- 4 months. The average age was 54 +/- 26 years. IOP, visual acuity and ocular complications were analysed. RESULTS: A total of 71 treatments were given. More than 1 treatment was given in 22 eyes (59%). The mean pretreatment IOP was 33.1 +/- 3.3 mm Hg. At the last follow-up, the mean IOP was 21 +/- 1.6 mm Hg. Mean total medications were reduced from 4.4 +/- 0.5 to 2.4 +/- 0.3. No serious complications occurred in 71 treatments. Visual acuity remained stable in 25 eyes (67.5%), improved in 6 eyes (16.2%) and decreased in 6 eyes (16.2%). CONCLUSIONS: DLPC is effective in lowering IOP in eyes with refractory glaucoma. It also serves to reduce the number of antiglaucoma medications. This procedure is relatively safe. Nevertheless, multiple DLPC applications may be needed. PMID- 11399933 TI - Intraocular pressure response to selective laser trabeculoplasty in the treatment of primary open-angle glaucoma. AB - PURPOSE: To evaluate the intraocular pressure (IOP) response to selective laser trabeculoplasty (SLT) in the treatment of uncontrolled primary open-angle glaucoma (POAG) in a prospective clinical study. SLT is a new laser procedure that selectively targets trabecular meshwork (TM) cells without coagulative damage to the TM. METHODS: 50 eyes with uncontrolled POAG were treated with a frequency-doubled, Q-switched Nd:YAG laser (532 nm). A total of approximately 50 nonoverlapping spots were placed over 180 degrees of the TM at energy levels ranging from 0.40 to 0.92 mJ/pulse. After SLT, the eyes were maintained with the identical hypotensive medical therapy as that before treatment. IOP was estimated before and 1 day, 7 days, 1 month, 3 months and 6 months after treatment. RESULTS: The mean pretreatment IOP was 22.48 mm Hg (SD 1.84). At the end of 1 month of follow-up IOP was reduced with a mean of 4.86 mm Hg (SD 2.38) or 21.6%; after 3 months, IOP was reduced with a mean of 5.66 mm Hg (SD 2.40) or 25.2%; at the end of 6 months of follow-up, IOP was reduced with a mean of 5.06 mm Hg (SD 2.37) or 22.5%. CONCLUSIONS: SLT is an effective method for lowering IOP in the treatment of POAG. PMID- 11399934 TI - Combined Phaco-Emulsification Cataract Extraction and Pars plana Vitrectomy without Intra-Ocular Lens Implantation. AB - PURPOSE: To report the indications, surgical technique and visual outcome of combined phaco-emulsification cataract surgery and three-port pars plana vitrectomy (CPPV) in patients unsuitable for primary intra-ocular lens (IOL) implantation. METHOD: A 4-year review in a tertiary referral hospital. RESULTS: Thirty patients (30 eyes) had CPPV without primary IOL insertion. The patients had a mean pre-operative logMAR acuity of 1.81 (SD 0.49) which improved to 1.50 (SD 0.72) postoperatively (p = 0.036). CONCLUSION: CPPV with primary IOL insertion is now an established surgical technique. This series suggests that patients with contra-indications to primary IOL insertion may also benefit from CPPV. PMID- 11399935 TI - The biocompatibility of silicone, fluorosilicone and perfluorocarbon liquids as vitreous tamponades. an ultrastructural and immunohistochemical study. AB - PURPOSE: To investigate the interactions occurring at the interface between some currently used vitreous tamponading substances and the ocular tissues, in an early follow-up after surgery. METHODS: Adult albino rabbit eyes underwent vitrectomy and were injected intravitreally with silicone oils at 1,000 or 3,000 cSt, fluorosilicone oil and perfluorodecalin. Different morphological techniques (light microscopy, scanning and electron microscopy, immunohistochemistry) were applied. RESULTS: All the tested materials, although non-toxic, penetrate the ocular tissues also at the anterior segment level, and in the long-term follow-up this can yield functional impairment. No massive inflammation has been detected in the zones in contact with the materials, but immunoglobulins and complement fractions are anyway present in the stroma of various tissues and around the droplets of emulsified materials, suggesting a local immune reaction. CONCLUSIONS: Data from this study confirmed that a permanent vitreous substitute, showing a perfect biocompatibility, still does not exist. The indication to remove the tamponading substance within 2 months after the surgery emerges from this investigation, confirming previous studies. Despite some negative features, silicone oil still appears the most biocompatible material for vitreous replacement. PMID- 11399936 TI - Thalidomide and prednisolone inhibit growth factor-induced human retinal pigment epithelium cell proliferation in vitro. AB - Thalidomide and prednisolone were recently introduced as treatment modalities in age-related macular degeneration (AMD). Growth factor-induced activation of retinal pigment epithelial (RPE) cells is a crucial event in this disease. The purpose was to examine the effect of thalidomide and prednisolone on growth factor-preactivated RPE cells. Human RPE cells were stimulated with 10 ng/ml platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), or vascular endothelial growth factor (VEGF) for 24 h. Afterwards, thalidomide (50 microg/ml) or prednisolone (100 ng/ml) were added for 24 h. RPE cell proliferation was determined by [3H]-thymidine incorporation. PDGF and bFGF significantly stimulated human RPE cell proliferation (p < 0.005), the value for VEGF stimulation was not significant (p = 0.3). The effect of the growth factors was diminished after addition of thalidomide and prednisolone (p < 0.005). The current study shows that the inhibitory properties of thalidomide and prednisolone remain even after growth factor activation of the cells. PMID- 11399937 TI - Cyclosporine effects on clinical findings and impression cytology specimens in severe vernal keratoconjunctivitis. AB - PURPOSE: To investigate the possible effects of topical cyclosporine eye drops 2% (CsA) on conjunctival immune cells obtained by impression cytology technique and to determine the clinical relevance of this effect if present. METHODS: Ten consecutive patients with severe, resistant, and active vernal keratoconjunctivitis (VKC) were included in this study. All of them were treated with 2% of topical CsA drops. Symptom and clinical scores and conjunctival impression cytology specimens were obtained both before and following treatment. Impression cytology specimens were stained with anti-CD8+, CD4+, CD45RA+, and CD23+ antibodies and the percentages of positive cells were counted. The results were compared and correlation analyses were performed between clinical and laboratory data. RESULTS: Significant reductions were observed with respect to the percentages of CD4+ and CD23+ cells in the conjunctival impression cytology specimens and clinical and symptom scores following treatment with topical CsA, while no change occurred in the percentages of CD8+ and CD45RA+ cells. The percentages of CD4+ and CD23+ cells showed good correlations with symptom and clinical scores. CONCLUSION: Topical CsA treatment is a very effective alternative in severe VKC cases in clinical ground and clinical efficacy of topical CsA treatment in severe, resistant VKC cases can be (at least partly) related to reduction of the CD23+ and CD4+ cell populations on the conjunctival surface. PMID- 11399938 TI - Duration of non-Insulin-dependent diabetes mellitus and the TNF-beta NcoI genotype as predictive factors in proliferative diabetic retinopathy. AB - The object of the study was to investigate the share of the polymorphisms I/D ACE, endothelin 1 4127G/A and TNF-beta NcoI in the susceptibility to proliferative diabetic retinopathy (PDR) in non-insulin-dependent diabetes mellitus (NIDDM). Genotypes were detected by polymerase chain reactions and determined in a set of 246 Caucasian NIDDM subjects with defined PDR status. The relevance of genotypes and clinical characteristics to the PDR occurrence was tested using multiple linear regression models and discrimination analysis. The best predictive value for PDR was given by a combination of two parameters - NIDDM duration and the TNF-beta genotype (p < 1.10(-6) and p = 1.10(-2), respectively) with a correct retrograde prediction of 82.6%. A comparison of the TNF-beta NcoI allele frequencies revealed no difference between NIDDM and nondiabetic subjects (n = 176), but a statistically significant difference was found between PDR and non-PDR NIDDM subjects (after a correction for the number of comparisons p = 0.03), allele beta2 being associated with PDR. Our results identified the allele variant TNF-beta2 being associated with PDR in NIDDM. Diabetes duration and the TNF-beta NcoI genotype were proven to significantly predict PDR occurrence. The TNF-beta2 allele could be regarded as a separate genetic risk factor that increases the relative incidence of PDR in patients with NIDDM. PMID- 11399939 TI - Penetration of amikacin into aqueous humor of rabbits. AB - Amikacin is an aminoglycoside antibiotic that has poor corneal penetration due to its hydrophilic properties. The purpose of this study was to compare and evaluate the penetration of amikacin sulfate into aqueous humor of the rabbit eye when applied by different routes and concentrations, namely 100 or 250 mg/ml topical fortified amikacin eye drops, 100 or 250 mg/ml amikacin-embedded soft contact lenses and 25 mg subconjunctival amikacin injection. One hour after application, amikacin was not detectable in any of the 100 mg/ml concentration groups. High levels of amikacin above the minimum inhibitory concentration for susceptible bacteria were detected when applied subconjunctivally and by 250 mg/ml topical fortified routes. Topical fortified amikacin 250 mg/ml reached the highest value in the aqueous (p < 0.05). Our results point out the poor corneal penetration of amikacin in standard concentrations from the intact rabbit cornea and that subconjunctival injections might provide satisfactory penetration. PMID- 11399940 TI - Effect of bradykinin on cultured bovine corneal endothelial cells. AB - PURPOSE: To clarify the effect of bradykinin on cytosolic free calcium mobilization and cell proliferation in cultured bovine corneal endothelial cells (BCEC). METHODS: The cytosolic free calcium concentration (Ca2+]i) was measured with the InCa(TM) Imaging System after the treatment of bradykinin (10(-11) to 10(-7) M) alone or with the pretreatments of EGTA, bradykinin receptor (Bk1 and Bk2) antagonists and an inhibition of phospholipase C (U-73122). Also, the effect of bradykinin on cell proliferation in BCEC was evaluated using cell counts. RESULTS: In BCEC, [Ca2+]i in the resting state was 87 +/- 9 nM. Bradykinin induced an increment of [Ca2+]i in a concentration-dependent manner and its 50% effective concentration was approximately 5 x 10(-11) M. A [Ca2+]i increment at 10(-8) M bradykinin was inhibited with the pretreatment of EGTA, an extracellular calcium chelator. U-73122 (5 x 10(-6) M) attenuated the bradykinin-induced [Ca2+]i increment. The pretreatment of HOE-140 (Bk2 antagonist) almost attenuated the bradykinin (10(-8) M)-induced [Ca2+]i increase, but des-Arg9-[Leu(8)] bradykinin (Bk1 antagonist) did not suppress it. To investigate the physiological effect of bradykinin, the effect of bradykinin on cell proliferation was studied. 10(-8) M of bradykinin produced a significant increase in cell numbers. This mitogenic effect of bradykinin was inhibited by the Bk2 antagonist. CONCLUSIONS: Bradykinin-induced stimulation of the signal transduction pathway in BCEC is coupled with the Bk2 type receptor. Furthermore, bradykinin produces the mitogenic effect in BCEC. PMID- 11399941 TI - Effect of trimetazidine on retinal ischemia/reperfusion injury in rats. AB - PURPOSE: To investigate the effect of trimetazidine (TMZ), an antioxidant agent, on the ischemia/reperfusion (I/R) injury in rat retina histopathologically. METHODS: The retinal I/R model was carried out by the 4-vessel occlusion method on Wistar albino rats. Twenty-one rats were divided into 7 groups, each comprising 3 rats. The animals in groups 1, 2 and 3 underwent 30 min of ischemia + 4 h of reperfusion and were treated by the administration of saline, TMZ before reperfusion and TMZ before ischemia, respectively. The animals in groups 4, 5 and 6 underwent 90 min of ischemia + 4 h of reperfusion and were treated in the same way as those in groups 1, 2 and 3, respectively. The 7th group was sham operated. RESULTS: Thirty and 90 min of ischemia followed by 4 h of reperfusion induced retinal injury in the rat retina. Histopathologically, the inner plexiform and inner nuclear layers were the most affected parts. TMZ was able to reduce almost all retinal I/R damage when administered before ischemia. A cytoprotective effect of TMZ was partly observed in those animals which were treated before reperfusion. CONCLUSION: TMZ seemed to have a protective effect against retinal I/R injury in rats. PMID- 11399942 TI - Orbital epithelial cyst derived from the lacrimal sac. AB - We describe a 32-year-old female who presented with a painless, moderately hard mass in the left inferior medial canthal region which had been present for about 6 years. She suffered from epiphora in the left eye in that time. The nasolacrimal system was completely patent on irrigation. Computed tomography showed a nodular mass with a diameter measuring about 25 mm located over the upper portion of the left lacrimal sac. The lesion was found to be a cyst located separately with close proximity to the lacrimal sac via anterior orbitotomy. Histopathologic examination revealed a cystic cavity lined by columnar to cuboidal epithelium consistent with a cyst derived from lacrimal sac epithelium. The patient has remained free of symptoms with a patent lacrimal system during the follow-up period of 1 year, without recurrence of the lesion. Orbital cysts of lacrimal sac derivation should be considered in the diagnosis of medial orbital tumors. Careful evaluation clinically with imaging studies and planning surgery constituted adequate therapy in this case. PMID- 11399944 TI - Expression of ETS-1 is correlated with urokinase-type plasminogen activator and poor prognosis in pulmonary adenocarcinoma. AB - Urokinase-type plasminogen activator (u-PA) is known to be involved in proteolytic degradation of extracellular matrix during carcinoma invasion. Similarly Ets-1, a transcription factor, is also known to be important for carcinoma progression, and has been reported to interact with the u-PA gene enhancer. We used immunohistochemistry to analyze the relationship between Ets-1 and u-PA protein expression in patients with pulmonary adenocarcinoma. A significant association was found between Ets-1 expression and u-PA expression. The Ets-1 expression was correlated with T factor, lymph node metastasis and stage. Kaplan-Meier survival analysis revealed that survival times were longer in patients with Ets-1-negative tumors than in patients with Ets-1-positive tumors. Our results suggest an important role for Ets-1 in tumor progression, and the correlation with u-PA protein expression serves as an important prognostic factor in pulmonary adenocarcinomas. PMID- 11399945 TI - Lactacystin inhibits cathepsin A activity in melanoma cell lines. AB - We describe the inhibitory effect of the proteasome inhibitor, lactacystin, on cathepsin A activity in murine melanoma cell lines. In vitro lactacystin metabolite, beta-lactone, at a concentration of 1 microM, significantly suppressed cathepsin A activity in B78 melanoma cell lysates by about 50%. Exposure of three murine melanoma cell lines with different metastatic potential to lactacystin at a concentration of 5 microM for 6 h caused a significant reduction in the carboxypeptidase activity of this enzyme, while the inhibitory activity remained unchanged for at least 12 h. Other proteasome-specific inhibitors, e.g. epoxomicin and N-benzyloxycarbonyl-Ile-Glu(O-tert-Bu)-Ala leucinal (PSI) at a concentration of 1 microM did not affect cathepsin A activity in melanoma cell line lysates. These data support our previous proposal that lactacystin is not a specific inhibitor of the proteasome. Since cathepsin A is also a tumor-associated enzyme, further research is needed to clarify its role and the significance of its inhibition by lactacystin in tumor biology. PMID- 11399946 TI - New immunoassays for MUC1 in breast cancer. AB - Eleven experimental immunofluorometric assays (IFMAs) were made using antibodies previously tested for epitope specificities. These assays were compared with six commercially available immunoassays. The clinical performance of these experimental assays was evaluated by analysing sera from 138 breast cancer patients and 105 female blood donors. The clinical performance of these assays was evaluated at a set specificity of 0.94. The highest overall sensitivity (0.56) was observed in the experimental assay with the antibody BC2 as solid phase and GP1.4 as the tracer antibody. This combination also showed the highest sensitivity in stage I/II breast cancer. The Truquant assay (Biomira) had an overall sensitivity of 0.51, and the highest sensitivity in stages III and IV at 0.65 and 0.94, respectively. The remaining commercial assays, with sensitivity ranging from 0.67 to 0.79, were below the top five experimental assays that showed sensitivity values between 0.79 and 0.85. The findings from our current study suggest that further development in MUC1 immunoassays could improve the detection of relapse in breast cancer patients. PMID- 11399947 TI - Elevated CA125 in breast cancer--A sign of advanced disease. AB - The serum tumor markers CA 125, MUC1 and CEA were measured in 221 breast cancer patients over a period of 2 years. Patients examined on at least three occasions were included in the study. Thirty-three patients had increasing or continuously high concentrations of CA 125. Thirty (91%) of these had involvement of the pleura, either as pleural metastasis or metastasis in surrounding tissue i.e. bone structures in the thorax cavity or lung parenchyma. MUC1 and CEA were elevated in 27 (82%) and 24 (73%) of the 33 patients, respectively. Increased concentrations of these two markers did not relate to the site of metastasis. However, the three tumor markers complemented each other in detecting early metastases. Increased CA 125 was associated with metastasis in or near the pleura, and in stage IV breast cancer it was related to poor prognosis. PMID- 11399948 TI - Effects of dextranation on the uptake of peptides in micrometastases: studies on binding of EGF in tumor spheroids. AB - Four different types of radiolabelled dextranated EGF were added to spheroids consisting of the human glioma cells U-343MGaCl2:6. Binding was analysed both in the peripheral well-nourished regions and in the deeper regions containing mainly quiescent cells. The substances had different molecular weights, and they were characterized regarding hydrophilic properties and isoelectric points. Two of the analysed conjugates, 125I-EGF-dextran (CDAP) and 125I-EGF-allyldextran-BSH, gave very low 125I binding in the deeper regions even after 24 h of incubation while better binding in these regions was found for 125I-EGF-dextran-DTPA, 125I-EGF allyldextran and for the reference substance 125I-EGF. The molecular weight seemed not to be of major importance for the binding properties and there were no clear relationships between binding and the hydrophilic properties or the isoelectric point values. The obtained differences could not be explained by differences in molecular weight or easily measured physicochemical parameters such as hydrophilic properties or isoelectric point values. Thus, other explanations must be found. PMID- 11399949 TI - Effects of vascular endothelial growth factor and E-selectin on angiogenesis in the murine metastatic RCT sarcoma. AB - The relationship between vascular endothelial growth factor (VEGF) and induction of angiogenesis in high-metastatic RCT(+) or low-metastatic RCT(-) clones of the poorly differentiated murine RCT sarcoma was investigated. The association with E selectin in VEGF-induced angiogenesis was also evaluated. RCT(+) cells produced significantly larger amounts of VEGF than RCT(-) cells. In a tube formation assay with murine lung microvascular endothelial (MLE) cells, conditioned medium from RCT(+) cells showed a significantly greater effect on tube formation than that from RCT(-) cells. Induction of tube formation was suppressed by anti-mouse VEGF monoclonal antibody. Furthermore, anti-mouse E-selectin monoclonal antibody suppressed the tube formation induced by recombinant mouse VEGF. In a flow cytometric analysis, the expression of E-selectin on MLE cells was upregulated after pretreatment with conditioned medium from RCT(+) and RCT(-) cells. Conditioned medium from RCT(+) cells induced a higher expression of E-selectin compared to medium from RCT(-) cells. Anti-VEGF monoclonal antibody prevented the upregulation of E-selectin by the RCT cell-conditioned medium. These findings suggest that E-selectin plays an important role in the angiogenesis induced by VEGF. VEGF derived from tumor cells may enhance angiogenesis by upregulating the expression of E-selectin on vascular endothelial cells. PMID- 11399950 TI - Retinoic acids reduce matrilysin (matrix metalloproteinase 7) and inhibit tumor cell invasion in human colon cancer. AB - All-trans retinoic acid (ATRA), 9-cis retinoic acid and 13-cis retinoic acid are naturally occurring retinoids used in the prevention and therapy of various preneoplastic and neoplastic diseases. It was previously reported that matrilysin, one of the matrix metalloproteinases (MMP-7), plays a critical role in the invasion and metastasis of gastrointestinal cancers. Moreover, it has been shown that ATRA downregulates matrilysin expression and prevents in vitro invasion by colon cancer cells. In this study, three retinoids were used, both in Matrigel invasion assays and in subcutaneous xenografts in mice, to evaluate the effects of retinoids on invasion by colon cancer cell lines (CHC-Y1, DLD-1, HT 29, BM314, CaR-1 and WiDr). All three retinoic acids tested reduced matrilysin expression and suppressed the invasiveness of colon cancer cell lines in vitro. Retinoic acids also reduced tumor invasion in mice without influencing tumor growth. Matrilysin expression in these tumors was clearly reduced. These data support the use of retinoic acids as useful reagents to manage patients with colorectal carcinoma. PMID- 11399951 TI - Analysis of epitopes of mouse monoclonal antibodies against human alpha fetoprotein. AB - Thirty-six monoclonal antibodies (MAbs) against human alpha-fetoprotein (AFP) were analyzed for the location of their epitopes by reacting them with a set of yeast recombinant AFP proteins using ELISA. Recombinant AFP proteins containing either one, two or all three domains, i.e. domain I, domain III, domain I-II, domain II-III and domain I-II-III, were produced and secreted into the culture medium of yeast cells harboring the expression plasmids. Epitopes of 13 MAbs were localized on domain I and 17 others were on domain III. However, the exact location of the epitopes of the remaining 6 MAbs could not be defined. The epitope of an antibody, namely AFY6, which was located in domain I, was successfully mapped on an octapeptide, C175KAENAVE182, using synthesized overlapping octapeptides. PMID- 11399952 TI - Absence of microsatellite instability and germline mutations of E-cadherin, APC and p53 genes in Japanese familial gastric cancer. AB - To evaluate the genetic factors of familial predisposition to gastric cancer, genetic alterations in the surgically resected stomach samples from gastric cancer-prone families were investigated. Familial gastric cancer (FGC) was defined as gastric cancer occurring in a family with 3 or more gastric cancer patients over at least two successive generations. We examined replication error (RER) of six microsatellite markers and screened mutations of the 10-(A) repeat sequence in the transforming growth factor-beta receptor type II (TGF-betaRII) gene in individuals from seven unrelated FGC families. Three cases showed RER at one of the six (CA)n microsatellite markers but the other 4 cases showed no RER at any of these loci. No mutation was found in the 10-(A) repeat of the TGF betaRII gene. Additionally, no germline mutation was found by polymerase chain reaction-single strand conformation polymorphism in exons 1-16 of E-cadherin, exons 5-8 of p53 and in the mutation cluster region of APC. These results indicate that disorders in the DNA mismatch repair system, E-cadherin, p53 and APC may be infrequently involved in the carcinogenesis of Japanese FGC. PMID- 11399953 TI - MUC1: antibodies and immunoassays. AB - High molecular weight mucins represent a unique challenge as tumor markers by virtue of their complex array of epitopes. The list is dominated by the high molecular weight mucins MUC1, CEA and CA125. While the currently accepted role for these tumor markers is in the prediction and detection of relapse, it is possible that their sensitivity and specificity can be improved. Although immunoassays detecting the tumor marker MUC1 are both sensitive and specific for predicting relapse in breast cancer, so far they are not in widespread use in the follow-up of this disease. Are there new combinations of conventional reagents that could improve assay sensitivity, or should we be looking for more radical changes in assay design incorporating combinatorial technology? PMID- 11399954 TI - Natural selection results in conservation of HIV-1 integrase activity despite sequence variability. AB - BACKGROUND: Integration of the HIV genome by integrase is absolutely required for productive infection. OBJECTIVE: To determine the role of natural selection on HIV integrase biology. DESIGN: To study the activities of HIV integrases from a limited panel of North American clinical isolates from HIV-infected patients and to compare these proteins with integrases from two laboratory adapted reference strains (HI(VIIIRF) and HIV(NL4--3)). METHODS: HIV was isolated and the particle associated RNA was reverse transcribed and sequenced. Replication kinetics of molecularly cloned viruses containing each variant integrase were studied in tissue culture. The mutant integrase proteins were expressed, purified and specific activities of the enzymes were derived for both 3' end-processing and disintegration reactions. RESULTS: Despite 3--5% variability in integrase at the amino acid level, viruses showed no statistically significant differences in growth kinetics compared with the reference HIV(NL4--3) virus and only minor differences were observed in 3' end-processing and disintegration activities. All integrase proteins demonstrated similar sensitivity to an integrase inhibitor l chicoric acid. CONCLUSIONS: These results demonstrate that integrase genes derived from HIV-infected individuals can differ from reference sequences but these mutations do not result in loss of function, including susceptibility to an integrase inhibitor; therefore, integrase remains an attractive target for antiviral drug design, as mutability appears to be restricted by function. PMID- 11399955 TI - Analysis of HIV-1 reverse transcriptase and protease sequences in paired plasma and lymphoid tissue specimens from HIV-1 infected individuals. AB - OBJECTIVES: To determine how representative the genotype of HIV-1 circulating in plasma is of the genotype of the virus present in lymphoid tissue. METHODS: Paired plasma and tonsillar tissue samples were prospectively obtained from patients with various levels of plasma HIV-1 RNA who were receiving combination antiretroviral therapy. HIV-1 reverse transcriptase and protease sequences were amplified from plasma and lymphoid tissue specimens by nested polymerase chain reaction and analyzed using an automated sequencing system. Results were compared with consensus HIV-1 sequences to determine whether drug-resistance mutations were present in the regions analyzed. RESULTS: HIV-1 protease sequences were compared in 11 plasma/tissue pairs obtained from eight patients; HIV reverse transcriptase sequences were compared in 12 plasma/tissue pairs obtained from nine patients. Sequence homology between plasma and tissue RNA, tissue RNA and DNA, and plasma and tissue DNA ranged from 97% to 100%. Few discrepancies were found when the percentage of mutant sequences at resistance codons was compared among paired samples. In most instances, tissue RNA or plasma contained a higher percentage of mutant sequences than did tissue DNA. CONCLUSION: The genotype of plasma HIV-1 is similar to the genotype of the virus in lymphoid tissue. Resistance studies using plasma samples should provide accurate information regarding the genotype of HIV-1 in lymphoid tissues. PMID- 11399956 TI - HIV in body fluids during primary HIV infection: implications for pathogenesis, treatment and public health. AB - OBJECTIVE: To describe initial viral dissemination to peripheral tissues and infectious body fluids during human primary HIV infection. DESIGN: Observational cohort study. METHODS: Blood plasma, cerebrospinal fluid (CSF), seminal plasma, cervicovaginal lavage fluid and/or saliva were sampled from 17 individuals with primary HIV infection (range of time from symptoms onset to sampling, 8--70 days) and one individual with early infection (168 days). Subjects' HIV-1 RNA levels in each fluid were compared with levels from antiretroviral-naive controls with established HIV infection. For study subjects, correlations were assessed between HIV-1 RNA levels and time from symptoms onset. Responses to antiretroviral therapy with didanosine + stavudine + nevirapine +/- hydroxyurea were assessed in each compartment. RESULTS: HIV-1 RNA levels were highest closest to symptoms onset in blood plasma (18 patients) and saliva (11 patients). CSF HIV-1 RNA levels (five patients) appeared lower closer to symptoms onset, although they were higher overall in primary versus established infection. Shedding into seminal plasma (eight patients) and cervicovaginal fluid (two patients) was established at levels observed in chronic infection within 3--5 weeks of symptoms onset. High-level seminal plasma shedding was associated with coinfection with other sexually transmitted pathogens. Virus replication was suppressed in all compartments by antiretroviral therapy. CONCLUSIONS: Peak level HIV replication is established in blood, oropharyngeal tissues and genital tract, but potentially not in CSF, by the time patients are commonly diagnosed with primary HIV infection. Antiretroviral therapy is unlikely to limit initial virus spread to most tissue compartments, but may control genital tract shedding and central nervous system expansion in primary infection. PMID- 11399957 TI - Increased risk of lipodystrophy when nucleoside analogue reverse transcriptase inhibitors are included with protease inhibitors in the treatment of HIV-1 infection. AB - BACKGROUND: Changes in body fat distribution are an adverse effect of therapy with HIV protease inhibitors (PI). It has been suggested that nucleoside analogue reverse transcriptase inhibitors (NRTI) may also contribute to this so-called lipodystrophy syndrome, but the relative contribution of the two drug classes is unclear as they are usually administered concomitantly. METHOD: The occurrence of lipodystrophy, as reported by physicians using no standardized criteria, was followed in patients randomly assigned to treatment with either a PI alone or a PI combined with an NRTI. The patients were part of a multicenter, open-label, randomized comparison of ritonavir (RTV)/saquinavir (SQV) with or without the addition of stavudine (d4T) in HIV-1-infected patients without prior PI and d4T experience (the Prometheus study). RESULTS: Lipodystrophy was reported in 29 of 175 (17%) patients during 96 weeks of follow up. Overall, it was reported significantly more frequently in patients who were randomized to RTV/SQV/d4T (22/88; 25%), than in patients randomized to RTV/SQV alone (7/87; 8%) (P = 0.003). When the analysis was limited to patients without any prior antiretroviral experience, lipodystrophy likewise was significantly more frequent in patients randomized to RTV/SQV/d4T (12/50; 24%) than in those randomized to RTV/SQV (2/44; 5%) (P = 0.008). CONCLUSION: This randomized clinical trial, in spite of not having been blinded, supports a contributory role of NRTI in the development of antiretroviral therapy-associated lipodystrophy. The low incidence of lipodystrophy in patients with no or limited NRTI exposure supports further evaluation of NRTI-sparing regimens as alternatives to current antiretroviral regimens. PMID- 11399958 TI - Recommendations for the clinical development of topical microbicides: an update. AB - Topical microbicides are products that are being developed to prevent HIV infection and other sexually transmitted diseases (STD) through topical application to the genital and rectal epithelial surfaces. This paper is an update of the clinical section of a general guidance for the development and evaluation of microbicidal products that was first published by the International Working Group on Microbicides (IWGM) in 1996. (The preclinical section of that document will be updated separately later.) All topical microbicides should be clinically evaluated in humans for safety and effectiveness. Safety studies are necessary to evaluate the potential for systemic absorption and toxicity as well as local toxic effects, such as irritation, ulceration, burning, and itching. Reported symptoms of burning and itching are relevant to future product use and acceptability. Irritation and ulceration of the vaginal, cervical, penile, or rectal epithelium have the potential to result in an increased transmission of HIV and other STD. Effectiveness studies to assess the prevention of HIV infection or STD, depending upon the product indication, are subsequently conducted. These trials need to be large enough to detect clinically meaningful levels of protection. For spermicidal microbicides, additional contraceptive effectiveness studies are also needed. PMID- 11399959 TI - Effect of early chemoprophylaxis with co-trimoxazole on nutritional status evolution in HIV-1-infected adults in Abidjan, Cote d'Ivoire. AB - BACKGROUND: In sub-Saharan Africa, malnutrition is a major complication of HIV disease. Measuring accurately the nutritional benefits of a therapeutic intervention could be an easy-to-monitor secondary outcome. METHODS: Anthropometric data were analysed from patients participating in a placebo controlled trial of co-trimoxazole prophylaxis in adults recruited at early stages of HIV-1 infection in Cote d'Ivoire (COTRIMO-CI ANRS 059 trial). Body mass index (BMI), arm muscle circumference (AMC) and percentage of fat mass (FM) were measured at baseline and quarterly during the follow up. Percentage of variation from the baseline value was compared between treatment groups and within the groups using Student t-test. RESULTS: An improvement of all anthropometric indicators was observed in the first 3 months of follow up in both treatment groups, significant in the co-trimoxazole group (P < or = 0.0006) but not in the placebo group (P > or = 0.06). In the co-trimoxazole group, this improvement was maintained for up to 24 months for BMI (P = 0.007), 21 months for AMC (P = 0.02) and only up to 12 months for FM (P = 0.04). The placebo group had a stable anthropometric status up to the end of the trial. Differences between treatment groups were significant for up to 15 months for BMI and AMC and 12 months for FM. CONCLUSION: As co-trimoxazole prophylaxis is now recommended in Africa as part of a minimum package of care for HIV-infected symptomatic subjects, the short-term improvement of these anthropometric indicators in adults who start co-trimoxazole prophylaxis should be considered as an effective clinical outcome. PMID- 11399960 TI - Concurrent sexual partnerships and HIV prevalence in five urban communities of sub-Saharan Africa. AB - OBJECTIVE: To estimate parameters of concurrent sexual partnerships in five urban populations in sub-Saharan Africa and to assess their association with levels of HIV infection and other sexually transmitted infections (STI). METHODS: Data were obtained from a multicentre study of factors which determine the differences in rate of spread of HIV in five African cities. Consenting participants were interviewed on sexual behaviour and at four of the five sites also provided a blood and a urine sample for testing for HIV and other STI. Data on sexual behaviour included the number of partnerships in the 12 months preceding the interview as well as the dates of the start and end of each partnership. Summary indices of concurrent sexual partnerships -- some of which were taken from the literature, while others were newly developed -- were computed for each city and compared to HIV and STI prevalence rates. RESULTS: A total of 1819 adults aged 15 -49 years were interviewed in Dakar (Senegal), 2116 in Cotonou (Benin), 2089 in Yaounde (Cameroon), 1889 in Kisumu (Kenya) and 1730 in Ndola (Zambia). Prevalence rates of HIV infection were 3.4% for Cotonou, 5.9% for Yaounde, 25.9% for Kisumu and 28.4% for Ndola, and around 1% for Dakar. The estimated fraction of sexual partnerships that were concurrent at the time of interview (index k) was relatively high in Yaounde (0.98), intermediate in Kisumu (0.44) and Cotonou (0.33) and low in Ndola (0.26) and in Dakar (0.18). An individual indicator of concurrency (iic) was developed which depends neither on the number of partners nor on the length of the partnerships and estimates the individual propensity to keep (positive values) or to dissolve (negative values) on-going partnership before engaging in another one. This measure iic did not discriminate between cities with high HIV infection levels and cities with low HIV infection levels. In addition, iic did not differ significantly between HIV-infected and uninfected people in the four cities where data on HIV status were collected. CONCLUSION: We could not find evidence that concurrent sexual partnerships were a major determinant of the rate of spread of HIV in five cities in sub-Saharan Africa. HIV epidemics are the result of many factors, behavioural as well as biological, of which concurrent sexual partnerships are only one. PMID- 11399961 TI - HIV infection among youth in a South African mining town is associated with herpes simplex virus-2 seropositivity and sexual behaviour. AB - OBJECTIVES: To determine the seroprevalence of HIV and herpes simplex virus-2 (HSV-2) by age and gender among young people aged 14--24 years in a South African town and to identify risk factors for HIV infection. DESIGN: A community-based, cross-sectional study was conducted on a random sample of men (n = 723) and women (n = 784) living in a township in the Carletonville district of South Africa. METHODS: Potential demographic and behavioural risk factors associated with HIV were recorded by questionnaire and biological tests were performed on serum and urine. Data analysis was performed using multivariate logistic regression. RESULTS: Among men and women the prevalence of HIV infection was 9.4 and 34.4%, respectively, and of positive HSV-2 serology was 17.0 and 53.3%, respectively. Among 24-year-old women the prevalence of HIV was 66.7% [95% confidence interval (CI), 54.6--77.3%]. HSV-2 seropositivity was a strong independent risk factor for HIV infection with odds ratios of 5.3 (95% CI, 2.7--10.3) for men and 8.4 (95% CI, 4.9--14.2) for women. There was no independent effect of age at first sex or serological markers of other sexually transmitted infections on HIV infection. CONCLUSIONS: HIV infection among young women increases rapidly after the onset of sexual activity and reaches extremely high levels by 24 years of age. These findings suggest that rates of HIV transmission from men to women are high and that HSV-2 plays a major role in the spread of HIV in this population. PMID- 11399962 TI - Increasing rates of malarial fever with deteriorating immune status in HIV-1 infected Ugandan adults. AB - BACKGROUND: Falciparum malaria and HIV-1 infection are two of the most important health problems facing sub-Saharan Africa. No convincing evidence of an association between symptomatic malaria and HIV-1 infection has been found. OBJECTIVE: To investigate the effect of HIV-associated immunosuppression on malarial fever rates. DESIGN: An observational cohort study in HIV-specific, primary healthcare clinics in Entebbe, Uganda, on 1371 HIV-1-infected adults participating in a randomized trial of 23-valent pneumococcal vaccine. METHODS: Cohort members underwent routine 6 monthly surveillance and had open clinic access when sick. Episodes of fever were assessed according to standardized protocols. Rates of malaria are described according to HIV immune status determined by CD4 T cell counts. RESULTS: Incidence rates of Plasmodium falciparum malarial fever showed a marked inverse relationship with CD4 T cell count; 140, 93 and 57 cases per 1000 pyo for CD4 T cell groups < 200, 200--499 and > 500 respectively, P < 0.001. Malarial fever definitions incorporating parasite density criteria (derived from asymptomatic surveillance) to correct for chance findings of fever and P. falciparum parasitaemia, did not affect the association of incidence rates with immunosuppression. CONCLUSION: These data support an interaction between symptomatic P. falciparum and HIV. Emphasis on mosquito avoidance measures should be an important component of education and counselling of HIV/AIDS patients in malaria-endemic areas, and suggests an additional HIV-related public health problem in Africa. PMID- 11399963 TI - Declining HIV prevalence and risk behaviours in Zambia: evidence from surveillance and population-based surveys. AB - OBJECTIVE: To examine trends in HIV prevalence and behaviours in Zambia during the 1990s. METHODS: The core Zambian system for epidemiological surveillance and research has two major components: (i) HIV sentinel surveillance at selected antenatal clinics (ANC) in all provinces; and (ii) population-based HIV surveys in selected sentinel populations (1996 and 1999). The former was refined in 1994 to improve the monitoring of prevalence trends, whereas the latter was designed to validate ANC-based data, to study change in prevalence and behaviour concomitantly and to assess demographic impacts. RESULTS: The ANC-based data showed a dominant trend of significant declines in HIV prevalence in the 15--19 years age-group, and for urban sites also in age-group 20--24 years and overall when rates were adjusted for over-representation of women with low education. In the general population prevalence declined significantly in urban women aged 15- 29 years whereas it showed a tendency to decline among rural women aged 15-24 years. Prominent decline in prevalence was associated with higher education, stable or rising prevalence with low education. There was evidence in urban populations of increased condom use, decline in multiple sexual partners and, among younger women, delayed age at first birth. CONCLUSIONS: The results suggested a dominant declining trend in HIV prevalence that corresponds to declines in incidence since the early 1990s attributable to behavioural changes. Efforts to sustain the ongoing process of change in the well-educated segments of the population should not be undervalued, but the modest change in behaviour identified among the most deprived groups represents the major preventive challenge. PMID- 11399964 TI - Using cost-effectiveness league tables to compare interventions to prevent sexual transmission of HIV. AB - Cost-effectiveness information is needed to help public health decision makers choose between competing HIV prevention programs. One way to organize this information is in a 'league table' that lists cost-effectiveness ratios for different interventions and which facilitates comparisons across interventions. Herein we propose a common outcome measure for use in HIV prevention league tables and present a preliminary league table of interventions to reduce sexual transmission of HIV in the US. Fifteen studies encompassing 29 intervention for different population groups are included in the table. Approximately half of the interventions are cost-saving (i.e. save society money, in the long run), and three-quarters are cost-effective by conventional standards. We discuss the utility of such a table for informing the HIV prevention resource allocation process and delineate some of the difficulties associated with the league table approach, especially as applied to HIV prevention cost-effectiveness analysis. PMID- 11399965 TI - Clinical testing of microbicides: a global research priority. PMID- 11399966 TI - To stem HIV in Africa, prevent transmission to young women. PMID- 11399967 TI - Primary genotypic resistance of HIV-1 to the fusion inhibitor T-20 in long-term infected patients. PMID- 11399968 TI - A zero prevalence of anti-HIV in blood donors in Gaza: how can it be sustained? PMID- 11399969 TI - Defective HIV-1 provirus found in peripheral T lymphocytes and granulocytes in an AIDS patient imply viral infection of progenitor cells. PMID- 11399970 TI - Increase of lipid plasma concentrations under protease inhibitor-containing regimens are not related to modifications of the tumour necrosis factor system. PMID- 11399971 TI - Indinavir hair concentration in highly active antiretroviral therapy-treated patients: association with viral load and drug resistance. PMID- 11399972 TI - Structured treatment interruption: a confusion in terminology. PMID- 11399974 TI - Access to HIV/AIDS care and treatment in the south of the world. PMID- 11399973 TI - Metabolic effects of indinavir in healthy HIV-seronegative men. AB - BACKGROUND: Therapy with HIV protease inhibitors (PI) has been associated with hyperglycemia, hyperlipidemia and changes in body composition. It is unclear whether these adverse effects are drug related, involve an interaction with the host response to HIV or reflect changes in body composition. METHODS: Indinavir 800 mg twice daily was given to 10 HIV-seronegative healthy men to distinguish direct metabolic effects of a PI from those related to HIV infection. Fasting glucose and insulin, lipid and lipoprotein profiles, oral glucose tolerance (OGTT), insulin sensitivity by hyperinsulinemic euglycemic clamp, and body composition were measured prior to and after 4 weeks of indinavir therapy. RESULTS: Fasting glucose (4.9 +/- 0.1 versus 5.2 +/- 0.2 mmol/l; P = 0.05) insulin concentrations (61.7 +/- 12.2 versus 83.9 +/- 12.2 pmol/l; P < 0.05), insulin : glucose ratio (12.6 +/- 1.7 versus 15.9 +/- 1.9 pmol/mmol; P < 0.05) and insulin resistance index by homeostasis model assessment (1.9 +/- 0.3 versus 2.8 +/- 0.5;P < 0.05) all increased significantly. During OGTT, 2 h glucose (5.1 +/- 0.4 versus 6.5 +/- 0.6 mmol/l; P < 0.05) and insulin levels (223.1 +/- 48.8 versus 390.3 +/- 108.8 pmol/l;P =0.05) also increased significantly. Insulin mediated glucose disposal decreased significantly (10.4 +/- 1.4 versus 8.6 +/- 1.2 mg/kg x min per microU/ml insulin; 95% confidence interval 0.6--.0;P < 0.01). There was no significant change in lipoprotein, triglycerides or free fatty acid levels. There was a small loss of total body fat (15.8 +/- 1.4 versus 15.2 +/- 1.4 kg;P = 0.01) by X-ray absorptiometry without significant changes in weight, waist : hip ratio, and visceral or subcutaneous adipose tissue by computed tomography. CONCLUSIONS: In the absence of HIV infection, treatment with indinavir for 4 weeks causes insulin resistance independent of increases in visceral adipose tissue or lipid and lipoprotein levels. PMID- 11399975 TI - Future access to HIV vaccines. Report from a WHO-UNAIDS Consultation, Geneva, 2-3 October 2000. AB - Results from the first phase III efficacy trial of an HIV vaccine will be available within the next 2-3 years. Thus, it is imperative to start planning now to address how any effective vaccines should be used. In the absence of definitive information on the characteristics of the first generation of HIV vaccines, the following assumptions were made: the vaccine will (i) have only low to moderate efficacy (on the order of 50%); (ii) not be inexpensive (on the order of 10 to 30 US $ per dose); (iii) require multiple doses; and, (iv) at least initially, be available in limited quantities. A vaccine with that profile would not be suitable for general use in all countries, and it might have to be initially targeted to populations at higher risk of HIV infection. These populations will differ from region to region, according to the epidemiological situation. In most high and middle income countries potential target groups for an initial HIV immunization programme would include intravenous drug users, gay men, commercial sex workers, and high-risk heterosexuals, as well as healthcare workers exposed to blood. In sub-Saharan Africa, future HIV immunization programmes might include larger segments of the population. In order to plan future vaccination programmes it is important to estimate the need (size of target population) and the demand (uptake in target populations) for future HIV vaccines. In addition to the public sector demand for an HIV vaccine (to be used in public health programmes), there will also be a private sector demand driven by the willingness and ability of individuals and employers to pay for the vaccine. HIV vaccines would need to be delivered as part of comprehensive HIV prevention packages, including behavioral and health promotion interventions. This would be especially important with vaccines of moderate efficacy, in order to prevent increased risk behavior among vaccine recipients. To avoid false expectations, the vaccine message would need to be recast as part of the total prevention strategy, rather than the "magic bullet" that people have come to expect. Initial deployment of HIV vaccines could proceed through targeted vaccination campaigns, drawing from experience with other vaccines. These campaigns would be complex and expensive, and would require full participation and collaboration from all levels of the community, as well as considerable strengthening of the infrastructures required for vaccine delivery. Current candidate vaccines in phase III trials may not be appropriate for much of Africa and South Asia, two areas most in need of an HIV vaccine. Credible international efforts (''push and pull" mechanisms) are needed to create incentives for the industry to develop vaccines for these regions. Feasible financing mechanisms may have to be established to cover the cost of production and delivery of vaccines, in order to ensure equitable access to HIV vaccines around the world. In parallel to the deployment of the initial vaccine, additional bridging studies and effectiveness trials may be needed to expand vaccine use. Research should also continue at an increased pace to develop new generations of more effective vaccines, especially vaccines appropriate to Africa. Achieving these goals will require real political commitment from government and international organizations, to be materialized in specific actions and budget allocations. The daunting challenge of making future effective vaccines accessible to all populations in need will require a sustained collaborative effort on the part of all parties involved. PMID- 11399976 TI - nef-deleted HIV-1 inhibits phagocytosis by monocyte-derived macrophages in vitro but not by peripheral blood monocytes in vivo. AB - OBJECTIVE: HIV-1 infection impairs a number of macrophage effector functions, but the mechanism is unknown. We studied the role of HIV-1 Nef in modulating phagocytosis by human monocytes and monocyte-derived macrophages (MDM). DESIGN AND METHODS: Using a flow cytometric assay, phagocytosis of Mycobacterium avium complex (MAC) by monocytes in whole blood of Sydney Blood Bank Cohort (SBBC) members infected with a nef-deleted (Delta nef) strain of HIV-1 was compared with that of monocytes from uninfected or wild-type (WT) HIV-infected subjects. The specific impact of Nef on phagocytosis by MDM was determined by either infecting cells in vitro with Delta nef strains of HIV-1 or electroporating Nef into uninfected MDM. RESULTS: MAC phagocytic capacity of monocytes from SBBC members was equivalent to that of cells from uninfected individuals (P = 0.81); it was greater than that of cells from individuals infected with WT HIV-1 (P < 0.0001), irrespective of CD4 counts and HIV viral load. In contrast, in vitro infection of MDM with either Delta nef or WT strains of HIV-1 resulted in similar levels of HIV replication and equivalent impairment of phagocytosis via Fc gamma and complement receptors. Electroporation of Nef into MDM did not alter phagocytic capacity. CONCLUSIONS: This study provides evidence demonstrating the complex indirect effect of Nef on phagocytosis by peripheral blood monocytes (infrequently infected with HIV-1) in vivo. Conversely, the fact that MDM infected with either Delta nef or WT HIV-1 in vitro (high multiplicity of infection) show comparably impaired phagocytosis, indicates that HIV-1 infection of macrophages can directly impair function, independent of Nef. PMID- 11399977 TI - Loss of memory (CD27) B lymphocytes in HIV-1 infection. AB - OBJECTIVES: The mechanisms of B-cell dysfunction during HIV-1 infection, including polyclonal B-cell activation, are poorly understood. We studied the phenotype and the functionality of peripheral memory B cells in HIV-1-infected subjects. DESIGN: The phenotype of B cells and the responsiveness to T-cell dependent activation in vitro were analysed in 36 HIV-1-infected and 34 healthy subjects. METHODS: Phenotyping of B and T cells was performed by FACS. IgG content was measured in plasma (by nephelometry) and cultures (by enzyme-linked immunosorbent assay) of B lymphocytes activated through CD40 or CD27 ligation. Expression of Fas and Fas ligand was performed by FACS on B-cell subpopulations from five HIV-1-infected and four uninfected subjects. RESULTS: The peripheral memory (CD27) B cells were significantly reduced in HIV-1-infected subjects. The amount of memory B cells was low in both drug-naive subjects and patients undergoing antiretroviral therapy. Ex vivo expression of CD70 (CD27 ligand) on T cells was significantly higher in HIV-1-infected subjects and inversely correlated with the frequency of memory B cells. In spite of the reduced number of memory B cells, in vitro spontaneous and activation-induced IgG secretion was higher in HIV-1-infected patients than in uninfected controls. The hyperactivation status of B lymphocytes in HIV-1-infected patients was further confirmed by the finding of upregulation of Fas and FasL expression on memory B cells. CONCLUSIONS: Memory B lymphocytes are depleted from peripheral blood in HIV-1-infected subjects. Our ex vivo findings suggest that persistent T-cell activation may contribute to loss of memory B cells through upregulation of Fas/FasL on these cells and terminal differentiation into plasma cells. PMID- 11399978 TI - Differential disappearance of inhibitory natural killer cell receptors during HAART and possible impairment of HIV-1-specific CD8 cytotoxic T lymphocytes. AB - BACKGROUND: Highly active antiretroviral therapy (HAART) is associated with a decrease in viral replication to undetectable levels and with an increase in CD4 T lymphocytes. Residual HIV-1 replication occurs together with incomplete recovery of cytotoxic CD8 T lymphocyte (CTL) numbers and function. We sought to determine whether expression of HLA class I-specific inhibitory natural killer receptors (iNKR) on the CTL of patients who had been treated successfully with HAART for 24 months could be involved, at least in part, in residual CTL functional inhibition. METHODS: Two-colour cytofluorometry was used to analyse the expression of six different iNKR including p58.1, p58.2, p70, p140, CD94/NKG2A and LIR1/ILT2 on the CD3, CD8 lymphocytes of eight patients with successful long-term suppression of viral replication before and after 3, 6 and 24 months of HAART. Healthy subjects were analysed as controls. HIV-1-specific cytotoxic activity was determined after 24 months of HAART in the presence and absence of iNKR-masking. RESULTS: No significant reduction of iNKR expression on CD8 T cells was observed by 6 months. Expression of p70 and p140 was inversely correlated with the increasing CD4 numbers. After 24 months CD8 T-lymphocytes expressing p58.1, p58.2, p70, p140 and CD94/NKG2A returned to levels indistinguishable from those of the healthy controls. A significantly increased proportion of CD8 CTL still expressed LIR1/ILT2, a receptor with broad HLA-class I specificity. Functional analysis of freshly separated cells revealed that the disruption of the interaction between LIR1/ILT2 and HLA-class I could partly restore HIV-1-specific lysis. CONCLUSIONS: A decrease in CD3CD8iNKR cells is observed beyond 6 months of HAART. In some patients functional impairment due to LIR1/ILT2 expression may persist even after 24 months of successful HAART. PMID- 11399979 TI - Increased abdominal visceral fat is associated with reduced bone density in HIV infected men with lipodystrophy. AB - OBJECTIVE: To examine the relationship between bone density and changes in regional and whole body composition in HIV-infected men with and without lipodystrophy. DESIGN: Cross-sectional, observational study of HIV-infected men with and without lipodystrophy and matched HIV-negative controls. SETTING: Tertiary care academic medical institution. PATIENTS: A total of 59 men, belonging to three different groups: HIV-positive men with lipodystrophy (n = 21), HIV-positive men without lipodystrophy (n = 20), and age-matched and body mass index-matched HIV-negative controls (n = 18). METHODS: Bone density, markers of bone turnover and indices of calcium metabolism were measured in all subjects. Quantitative computed tomography was used both to determine volumetric bone density of the spine and to quantify abdominal visceral fat. Dual energy X-ray absorptiometry was used to determine whole body composition and bone density. Statistical comparisons were performed according to lipodystrophy categorization and protease inhibitor exposure. RESULTS: Men with HIV-associated lipodystrophy had reduced lumbar spine bone density compared with both HIV-infected non lipodystrophic men [mean +/- SD, 132 +/- 29 versus 154 +/- 30 mg/cm(3); P = 0.02] and HIV-negative controls [mean +/- SD 132 +/- 29 versus 148 +/- 18) mg/cm(3); P = 0.04]. Lumbar spine bone density was reduced significantly in HIV lipodystrophy patients independently of protease inhibitor use. In an analysis among all HIV infected subjects, increased visceral abdominal fat area was associated with decreased lumbar spine bone density (r, -0.47; P = 0.002). The association between visceral fat and bone density remained significant (P = 0.007) after controlling for age, body mass index, lowest body weight, protease inhibitor use, and extremity fat in a multivariate regression model. Markers of bone turnover were not related to bone density or lipodystrophy status. CONCLUSIONS: Lumbar spine bone density is reduced in association with increased visceral fat in HIV infected men with lipodystrophy. Further studies are needed to determine the mechanisms of osteopenia in HIV lipodystrophy and whether increased marrow fat occurs in such patients and affects bone density. PMID- 11399980 TI - When to start highly active antiretroviral therapy in chronically HIV-infected patients: evidence from the ICONA study. AB - OBJECTIVES: To compare the response to highly active antiretroviral therapy (HAART) in individuals starting HAART at different CD4 cell counts. DESIGN: The mean increase in CD4 cell count and rate of virological failure after commencing HAART were measured in antiretroviral-naive patients (1421) in a large, non randomized multicentre, observational study in Italy (ICONA). Clinical endpoints were also evaluated in a subset of patients who started HAART with a very low CD4 cell count. RESULTS: After 96 weeks of therapy, the mean rise in CD4 cell count was 280, 281 and 186 x 10(6) cells/l in patients starting HAART with a CD4 cell count < 200, 201--350 and > 350 x 10(6) cells/l, respectively. Patients starting HAART with a CD4 cell count < 200 x 10(6) cells/l tended to have a higher risk of subsequent virological failure [relative hazard (RH), 1.15; 95% confidence interval (CI), 0.93--1.42] compared with patients starting with > 350 x 10(6) cells/l. There was no difference in risk between the 201--350 and the > 350 x 10(6) cells/l groups (RH, 1.0; 95% CI, 0.79--1.29). The incidence of new AIDS defining diseases/death in patients who started HAART with a CD4 count < 50 was 0.03/person-year (95% CI, 0.10--0.33) during the time in which the patient's CD4 cell count had been raised to > 200 x 10(6) cells/l. CONCLUSIONS: There was no clear immunological or virological advantage in starting HAART at a CD4 cell count > 350 rather than at 200--350 x 10(6) cells/l. The increase in CD4 cells restored by HAART is meaningful in that they are associated with reduced risk of disease/death. PMID- 11399981 TI - Analysis of variation in plasma concentrations of nelfinavir and its active metabolite M8 in HIV-positive patients. AB - OBJECTIVE: To characterize sources of variation in plasma concentrations of nelfinavir and its active metabolite M8 and to evaluate the use of therapeutic drug monitoring for nelfinavir treatment. METHODS: Plasma samples and patient's characteristics were obtained from outpatient clinic. Differences between groups of patients were studied by comparing the observed plasma concentrations with the corresponding concentration on a pharmacokinetic population curve based on median plasma levels. RESULTS: Plasma samples (618) were available from 355 patients taking 1250 mg nelfinavir twice daily. The median ratio between M8 and nelfinavir concentrations was 0.29. This ratio appeared to be independent of the time after ingestion. Statistically significantly lower M8 concentrations were found in Black and Asian patients, or when comedication with CYP3A4 inducers was used. Coadministration of CYP2C19 inhibitors, such as omeprazole, decreased the median M8/nelfinavir ratio. Nevertheless, nelfinavir concentrations and summed concentrations of nelfinavir and M8 were only marginally affected in these patients. Diarrhoea was identified as a cause for lower nelfinavir concentrations, without changing the M8/nelfinavir ratio. In a number of patients with suspected therapy failure or intoxication, abnormal nelfinavir plasma concentrations were found. Dose adjustments based on nelfinavir plasma levels were helpful in a number of patients. CONCLUSION: This study shows that the total concentration of nelfinavir and M8 together is not significantly influenced when variation in M8 levels occurs. Consequently, measuring M8 concentrations in addition to nelfinavir concentrations is not required for the purpose of therapeutic drug monitoring for this drug. PMID- 11399982 TI - Clinical outcome among HIV-infected patients starting saquinavir hard gel compared to ritonavir or indinavir. AB - OBJECTIVE: To compare the clinical response among patients who initiate protease inhibitor therapies with different virological potency. DESIGN: We analysed patients who started indinavir, ritonavir or saquinavir hard gel capsule (hgc) as part of at least triple therapy during prospective follow-up within the EuroSIDA study. METHODS: Changes in plasma viral load (pVL) and CD4 cell count from baseline were compared between treatment groups. Time to new AIDS-defining events and death were compared in Kaplan--Meier models, and Cox models were established to further assess differences in clinical progression (new AIDS/death). Adjustment was made for differences in baseline parameters, in particular pVL, CD4 cell count, and region of Europe. RESULTS: A total of 2708 patients (median follow-up: 30 months) were included, of which 556 started ritonavir (21%), 1342 indinavir (50%), and 810 saquinavir hgc (30%). The three groups were fairly evenly balanced at baseline regarding CD4 count, previous diagnosis of AIDS and pVL, After 12 months, the median changes in CD4 cell count were 90, 96 and 74 x 10(6) cells/l, respectively;P < 0.001, the proportions of patients with pVL < 500 copies/ml were 47, 54 and 41%; P < 0.001, and the proportions with clinical progression were 11.9, 9.2 and 11.9%, respectively; P = 0.20 (log-rank test). In multivariate models the relative risk of clinical progression for indinavir compared with saquinavir hgc was: 0.77 (0.60--0.99); P = 0.043, and for ritonavir 0.83 (0.62--1.11); P = 0.20. CONCLUSIONS: Saquinavir hgc was associated with an inferior long-term clinical response relative to indinavir, which was consistent with the observed differences in virological and immunological responses. PMID- 11399983 TI - Pharmacokinetics and safety of amprenavir and ritonavir following multiple-dose, co-administration to healthy volunteers. AB - OBJECTIVE: To evaluate the safety and pharmacokinetic interaction between amprenavir (APV) and ritonavir (RTV). METHODS: Three open-label, randomized, two sequence, multiple-dose studies having the same design (7 days of APV or RTV alone followed by 7 days of both drugs together) used 450 or 900 mg APV with 100 or 300 mg RTV every 12 h with pharmacokinetic assessments on days 7 and 14. Safety was monitored as clinical adverse events (AEs) and laboratory abnormalities. RESULTS: Relative to APV alone, RTV co-administration resulted in a 3.3- to 4-fold and 10.84 to 14.25-fold increase in the geometric least-square (GLS) mean area under the plasma concentration--time curve (AUC(tau,ss)) and minimum concentration (C(min,ss)), respectively. APV 900 mg with RTV 100 mg resulted in a 2.09-fold and 6.85-fold increase in the GLS mean AUC(tau,ss) and C(min,ss), respectively. On day 14, the geometric mean (95% confidence interval) for 450 mg APV AUC(tau,ss) (micro x h/mL) was 23.49 (19.32--28.57) with 300 mg RTV and 35.42 (30.46--44.42) with 100 microg RTV, and for the 900 mg APV with 100 mg RTV 47.11 (39.47--61.24). The 450 mg APV C(min,ss) (microg/ml) were 1.32 (1.05 -1.67) and 2.01 (1.70--2.61), and 2.47 (2.08--3.32) for 900 mg APV. The most common AEs were mild and included diarrhea, nausea/vomiting, oral parasthesias, and rash. The triglyceride and cholesterol increased significantly from RTV exposure. CONCLUSION: Adding RTV to APV resulted in clinically and statistically significant increases in APV AUC and C(min) with variable effects on maximum concentration. The two RTV doses had similar effects on APV but AEs were more frequent with 300 mg RTV. PMID- 11399984 TI - Sexual dysfunction associated with protease inhibitor containing highly active antiretroviral treatment. AB - BACKGROUND: A high proportion of individuals receiving highly active antiretroviral treatment (HAART) complain of sexual dysfunction (SD), encompassing a lack of desire or erectile dysfunction. OBJECTIVE: To determine whether SD was associated with particular components of the HAART regimens and to identify risk factors for the development of SD in patients on HAART. METHODS: A survey among patients with HIV infection using an anonymous questionnaire was conducted in 10 European countries between December 1998 and December 1999. A total of 904 individuals currently receiving antiretroviral agents were included in the analyses. RESULTS: A decrease in sexual interest was significantly more frequently reported by subjects (men and women) using HAART containing protease inhibitors (PI) (308/766, 40%), compared with PI-naive patients (22/138, 16%; OR 3.55; 95% CI 2.15--5.89). In addition, a significantly larger number of PI experienced men reported a decrease in sexual potency (216/628, 34%) compared with PI-naive men (12/99, 12%; OR 2.56; 95% CI 1.33--5.03). In multivariate analyses the following factors were associated with a decrease in sexual interest: a current PI-containing regimen, a history of a PI regimen, symptomatic HIV infection, age and homosexual contact as HIV transmission mode. Factors associated with a decrease in sexual potency were: current use of a PI-containing regimen, symptomatic HIV disease, age and the use of tranquillisers. CONCLUSION: SD appears to be a common side-effect of HAART regimens containing a PI. The potential association between SD and other side-effects of HAART, such as lipodystrophy syndrome and neuropathy, should be investigated further. PMID- 11399985 TI - Management of the risk of HIV infection in male homosexual couples. AB - OBJECTIVES: To study the ways of managing HIV risk within male homosexual steady relationships (gay couples), including factors associated with consistent condom use during anal sex with the steady partner. METHOD: An anonymous and standardized questionnaire completed by a convenience sample of homosexuals in Switzerland in 1997 (n = 1097). Information on the couple was provided by the 74% (n = 786) of male respondents who reported having a steady partner in the past 12 months. Data were analysed by contingency tables and logistic regression. RESULTS: Different ways of managing HIV risk were reported: negotiated safety (both HIV negative, condoms abandoned) was chosen by one quarter of the couples, but the most frequent solution was reliance on condoms for anal sex, chosen by more than four in 10. Altogether 84% of couples exhibited safe management of HIV risk within their partnership. The 16% of couples showing inadequate management of HIV risk within the couple mostly relied on questionable assumptions about past or present risks. A total of 74% of couples had spoken about managing HIV risk with possible casual partners. Reported behaviour with the steady partner and with casual partners was highly consistent with claimed strategies chosen to manage HIV risk. Consistent condom use with the steady partner was mostly associated with variables characterizing the relationship: initial 2 years of the relationship, discordant or unknown serological HIV status, non-exclusivity. CONCLUSION: Gay couples manage HIV risk in a variety of ways. Most strategies provide adequate protection with casual partners, but leave gaps in protection between the steady partners themselves. PMID- 11399986 TI - Female condom introduction and sexually transmitted infection prevalence: results of a community intervention trial in Kenya. AB - OBJECTIVE: To measure the impact on sexually transmitted infection (STI) prevalence of a female condom introduction and risk-reduction program at Kenyan agricultural sites. DESIGN: We conducted a cluster-randomized trial to determine whether a replicable, community-level intervention would reduce STI prevalence. METHODS: Six matched pairs of tea, coffee and flower plantations were identified. The six intervention sites received an information/motivation program with free distribution of female and male condoms, and six control sites received only male condoms and related information. Participants were tested for cervical gonorrhea and chlamydia by ligase chain reaction on urine specimens, and vaginal trichomoniasis by culture, at baseline, 6 and 12 months. RESULTS: Participants at intervention (n = 969) and control sites (n = 960) were similar; baseline STI prevalence was 23.9%. Consistent male condom use was more than 20% at 12 months. Consistent female condom use was reported by 11 and 7% of intervention site women at 6 and 12 months. Unadjusted STI prevalence was 16.5 and 17.4% at 6 months, and 18.3 and 18.5% at 12 months, at the intervention and control sites, respectively. Logistic regression models confirmed the null effect of the female condom intervention. CONCLUSIONS: Female condom introduction did not enhance STI prevention at these sites. It is unclear which aspects of the intervention -- STI education, condom promotion, case management -- were associated with decreased STI prevalence from baseline to follow-up. PMID- 11399987 TI - Positive and negative life events after counselling and testing: the Voluntary HIV-1 Counselling and Testing Efficacy Study. AB - INTRODUCTION: The results of the Voluntary HIV-1 Counselling and Testing Efficacy Study support the efficacy and cost-effectiveness of HIV voluntary counselling and testing (VCT) for reducing risk behaviors in three developing countries. METHODS: This report explores the social consequences of HIV VCT by examining the incidence of positive and negative life events at the first follow-up (an average of 7.3 months after recruitment). The incidence of positive and negative life events was compared between: (i) those who were randomly assigned to HIV VCT versus health information (HI); (ii) those who tested seronegative and those who tested seropositive; and (iii) those who disclosed their serostatus and those who did not. RESULTS: The occurrence of most negative life events was rare (0--4%); positive life events were more common (17--39%). With few exceptions, those assigned to HIV VCT were no more likely to experience negative life events than those who were assigned to HI. For individuals, positive serostatus was associated with increased support from health professionals, the break-up of a marriage and being neglected or disowned by their family. Serodiscordant couples with an HIV-positive woman were most likely to report the break-up of a marriage (20 versus 0--7% for other groups) and the break-up of a sexual relationship (45 versus 22--38% for other groups). Disclosure was associated with strengthening of a sexual relationship except for HIV-seropositive women. CONCLUSION: These findings, in combination with findings supporting the efficacy and cost effectiveness of HIV VCT, support the dissemination of HIV VCT with appropriate support services in developing countries. PMID- 11399988 TI - Gay men, risk and relationships. PMID- 11399989 TI - Editorial comment on Analysis of variation in plasma concentrations of nelfinavir and its active metabolite M8 in HIV-positive patients. PMID- 11399990 TI - Is long postpartum sexual abstinence a risk factor for HIV? PMID- 11399991 TI - Sperm mitochondrial DNA deletions as a consequence of long term highly active antiretroviral therapy. PMID- 11399992 TI - First CRF01_AE/B recombinant of HIV-1 is found in Thailand. PMID- 11399993 TI - MIP-1 alpha promoter polymorphism in humans and monkeys: identification of two polymorphic regions characterized by the insertion of unique sequences in monkeys. PMID- 11399994 TI - Prevention of invasive aspergillosis in AIDS by sulfamethoxazole. PMID- 11399995 TI - Long-term outcomes of protease inhibitor-based therapy in antiretroviral treatment-naive HIV-infected injection drug users on methadone maintenance programmes. PMID- 11399996 TI - Persistence of zidovudine-resistance mutations in HIV-1 isolates from patients removed from zidovudine therapy for at least 3 years and switched to a stavudine containing regimen. PMID- 11399997 TI - Lactic acid levels in children perinatally treated with antiretroviral agents to prevent HIV transmission. PMID- 11399998 TI - 'Do HIV-infected injecting drug users over-report adherence to highly active antiretroviral therapy?' A comparison between patients' self-reports and serum protease inhibitor concentrations in the French Manif 2000 cohort study. PMID- 11399999 TI - Low incidence of genotypic and phenotypic resistance in paediatric HIV-infected patients on long-term first-line antiretroviral triple therapy. PMID- 11400000 TI - Is indinavir crystalluria an indicator for indinavir stone formation? PMID- 11400001 TI - Homocysteinaemia in HIV-infected patients treated with highly active antiretroviral therapy. PMID- 11400002 TI - Prevalence of antiretroviral resistance in a South London cohort of treatment naive HIV-1-infected patients. PMID- 11400003 TI - Efavirenz-induced photoallergic dermatitis in HIV. PMID- 11400004 TI - Slower decline of plasma HIV-1 RNA after highly active antiretroviral therapy in primary versus chronic infection. PMID- 11400005 TI - AIDS conference world treatment. PMID- 11400017 TI - Prion disease resembling frontotemporal dementia and parkinsonism linked to chromosome 17. AB - OBJECTIVE: To compare the clinical features of a familial prion disease with those of frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP 17). BACKGROUND: Prion diseases are not usually considered in the differential diagnosis of FTDP-17, since familial Creutzfeldt-Jakob disease (CJD), the most common inherited prion disease, often manifests as a rapidly progressive dementia. Conversely, FTDP-17 usually has an insidious onset in the fifth decade, with abnormal behavior and parkinsonian features. METHOD: We present the clinical features of 12 patients from a family with CJD associated with a point mutation at codon 183 of the prion protein gene. RESULTS: The mean age at onset was 44.0 +/- 3.7; the duration of the symptoms until death ranged from two to nine years. Behavioral disturbances were the predominant presenting symptoms. Nine patients were first seen by psychiatrists. Eight patients manifested parkinsonian signs. CONCLUSION: These clinical features bear a considerable resemblance to those described in FTDP-17. PMID- 11400018 TI - Multiple sclerosis: report on 200 cases from Curitiba, Southern Brazil and comparison with other Brazilian series. AB - We reviewed the clinical and laboratory findings of 200 patients in Curitiba, Southern Brazil (25 degrees 25'40" S; 49 degrees 16'23" W-GR), with multiple sclerosis (MS)according to Poser's criteria. The patients were classified as: clinically definite (A1 and A2) - 142 patients (71%); laboratory-supported definite - 42 patients (21%); and clinically probable - 16 patients (8%). Relapsing-remitting (RR) form was the most common clinical presentation, with 182 (91%), followed by primary progressive (PP)(16 cases, 8%), and only 2 cases with secondarily progressive form (SP). Nine women and 7 men totalized the 16 PP cases. The mean age of onset was 32.0+/-9.9 (median 32 years). The gender ratio was female 1.8:1 male. All patients, except 3 African-Brazilian, were white. Seven (3.5%) patients developed a clinical history of Devic's syndrome. The initial clinical picture included brainstem/cerebellar syndrome in 126 (63%) cases, sensorial findings in 106 (53%)patients, motor (pyramidal) syndrome in 102 (49.5%), and optic neuritis in 79 (39.5%) cases. 122 (61%) patients had a final EDSS score < 3.5; 45 (22.5%) a score between 3.5 and 5.5, and 33 (16.5%) a score > or = 6.0. There was no significant correlation between the number of relapses or duration of disease with EDSS scores (Spearman's test). Only 14 (7%) of the total number presented the benign form (EDSS< 3.5 after 10 years of disease). We observed a later age of onset and initial clinical findings with higher frequency of brainstem/cerebellar syndrome and optic neuritis, when compared to other Brazilian and Western series PMID- 11400019 TI - Encoding process in delayed recall impairment and rate of forgetting in Alzheimer's disease. AB - OBJECTIVE: To assess the role of impaired encoding in learning and in delayed recall disturbances, and to evaluate the rate of forgetting in AD. METHOD: Fifteen AD patients with mild or moderate dementia and 15 normal matched controls were assessed with the Buschke Selective Reminding Test. Delayed recall was evaluated after 30 minutes and after 24 hours. RESULTS: AD patients had a poorer performance across the six trials of the learning phase as well as in both delayed recall evaluations, with no difference between recall at 30 minutes and at 24 hours. CONCLUSION: Performance in the learning phase was as specific and almost as sensitive as the performance in delayed recall for AD diagnosis. Encoding impairment was responsible for poorer learning and rapid displacement of previous learned material in the AD group. Finally, we did not find a higher rate of forgetting in AD patients. PMID- 11400020 TI - Validity and reliability of the Portuguese version of the Confusion Assessment Method (CAM) for the detection of delirium in the elderly. AB - This study has tested the validity and reliability of the Portuguese version of the Confusion Assessment Method (CAM), a diagnostic assessment instrument for delirium developed by Inouye et al. (1990). The sample was formed by 100 patients with 60 and more years of age, admitted at the emergency service of Santa Casa de Sao Paulo, in the time periods between July and August, 1996, November and December, 1996 and February and March, 1997. The sensibility was 94.1% and specificity 96.4%. The assessors reliability in a sample of the 24 patients resulted in a kappa = 0.70. We have concluded that CAM is an adequate instrument to assess the presence of delirium, reliable to assess elderly patients at the emergency services. PMID- 11400021 TI - Association between mental health screening by self-report questionnaire and insomnia in medical students. AB - Epidemiological research points to the high prevalence of psychiatric disorders among insomniacs. We carried out a cross-sectional study with medical students with the aim of evaluating the association between insomnia and suspicion of psychiatric disorder; 302 medical students were included (184 males and 118 females; mean age = 20.47+/-1.89 years). The main association was tested by logistic regression analysis. The overall prevalence of positivity in a screening test for psychiatric disorder was 22.19%; and of insomnia, 28.15%. Difficulty initiating sleep (OR=3.45), difficulty maintaining sleep (OR=7.61), falling asleep later (OR=1.99) and waking up earlier (OR=1.91) were associated with suspicion of psychiatric disorder. As a group, the variables difficulty initiating sleep, difficulty maintaining sleep, falling asleep after 11 pm, and waking up before 6 am presented an odds ratio of 5.96 for positivity in the screening for psychiatric disorder. Furthermore, difficulty maintaining sleep (OR=2.24) was associated with "being female," and falling asleep later (OR=0.43) was associated with "being male". These results underscore the importance of determining in what cases difficulty sleeping may have severe clinical repercussions or affect performance. PMID- 11400022 TI - Electrophysiological evaluation in myotonic dystrophy: correlation with CTG length expansion. AB - In myotonic dystrophy (MD), disease severity has been correlated with expansion of CTG repeats in chromosome 19. The aims of this study were to evaluate efficacy of electromyography in the diagnosis of MD, access the frequency and the characteristics of peripheral involvement in the disease and to verify whether the CTG repeats correlated with the electrophysiological abnormalities. Twenty five patients and six relatives at risk of carrying the MD gene were examined. Electrical myotonia (EM) was scored. Sensory and motor conduction velocity (CV) were studied in five nerves. Leukocyte DNA analysis was done in 26 subjects. Myopathy and myotonia were found in 27 cases. EM was most frequent in muscles of hand and in tibialis anterior. No significant correlation was found between EM scores and length of CTG expansions. EM scores correlated significantly with the degree of clinical myopathy, expressed by a muscular disability scale. Peripheral neuropathy was found in eight subjects and was not restricted to those who were diabetics. PMID- 11400023 TI - [A review of technical and physiological aspects of F-wave studies and analysis of the data obtained in a group of diabetic patients]. AB - We reviewed some physiological aspects of the F-wave studies, mainly related to the motoneurone sizes involved in the generation of this potentials and the number of stimuli necessary to analyze the F-wave parameters. F-wave latencies and F-wave conduction velocities obtained in a group of normal volunteers and in a group of diabetic patients are analyzed. PMID- 11400024 TI - [The cognitive potential in normal adults]. AB - The objective of this study was to evaluate the results of the auditory cognitive potential in a group of normal adults. The evoked response was obtained at Cz and Fz. The data collected were statistically analysed. The mean latencies of N1, P2, P2AP, N2, P3 and amplitude of P3, as well the standard deviation of these variables were presented. We briefly comment on some pitfalls related to interpretation of the results as well as the limitations of the technique as a diagnostic tool. PMID- 11400025 TI - [Prevalence and semiologic aspects of the idiopathic stabbing headache in a migraine population]. AB - Idiopathic stabbing headache is a quite unknown disorder. Its main features consist of brief stabbing pains, lasting few seconds. In most cases, idiopathic stabbing headache is underdiagnosed. We have followed up migraine patients during a period of 12 months, aiming to determine the prevalence and main features of idiopathic stabbing headache while occurring apart from migraine attacks. Two hundred and thirty-three of the 280 patients initially surveyed were included in the analysis of the results. Ninety-four patients presented idiopathic stabbing headaches (40.4%), being 72 of them females (76.5%). For migraine with idiopathic stabbing headaches, mean age, age of beginning of migraine and years with migraine were 33, 22.5 and 10.6 years, respectively. Mean duration of the idiopathic stabbing headaches was reported to be 1.42 seconds [ 1 second by 68 patients (72.4%), 2 seconds by 17 (18.1%), 3 seconds by 6 (6.3%), 4 seconds by 1 (1.05%) and 5 seconds by 2 (2.15%)]. Pain paroxysms were reported to be unilateral by 86 (91.4%) and bilateral by 8 (8.6%) of the cases. They were reported to be temporal by 56 patients (60%), occipital by 15 (15.6%), frontal by 8 (8.5%), temporo-occipital by 7 (7.4%), parietal by 5 (5.3%), fronto-temporal by 1 (1.06%), cervical by 1 (1.06%) and ocular by 1 patient (1.06%). The study confirms a high prevalence of idiopathic stabbing headaches in migraineurs. Its main clinical features could be well determined during the interval between migraine attacks. PMID- 11400026 TI - [Dilatation of Virchow-Robin spaces in patients with migraine]. AB - The Virchow-Robin spaces (VRS) are subpial invaginations making a tubular revestment scabbard that contains a vase between the subarachnoid and subpial spaces. The magnetic resonance imaging (MRI) is the only imaging method able for evaluate this anatomic detail. We studied the possible association between dilated VRS and migraine. Seventy patients with ages ranging from 13 to 54 years (mean, 36.5 years), with clinical diagnosis of migraine were studied and compared with a control group, without past of headache, composed by the same number of patients with ages ranged from 14 to 64 years (mean, 42 years). We observed widening of VRS in 28 cases (40%) of the migraine patients. In the control group this feature was demonstrated in only 5 cases (7.1%). Alert for the importance of detecting and recognizing the VRS, as well as, the differentiation from lacunar infarctions and choroid fissure cysts. PMID- 11400027 TI - [EPI-FLAIR sequence in the evaluation of mesial temporal sclerosis]. AB - The purpose of this study is to evaluate morphological and signal intensity changes in the hippocampus in patients with medically intractable temporal lobe epilepsy. We studied 8 patients with mesial temporal sclerosis using a 1.5 -T MR and the following sequences Spin Eco- SE, Fast Spin Echo- FSE, Fluid Attenuation Inversion Recovery Echo Planar Imaging--FLAIR-EPI. We noticed a sensitive increase signal intensity on FLAIR- EPI sequences, particularly, in coronal images, than on SE and FSE sequences. The STIR sequence showed a cortical hippocampus atrophy in half of the cases, in whom signal abnormalities were present. PMID- 11400028 TI - [The most common conditions in a neurology specialty clinic]. AB - OBJECTIVE: To present the most frequent diagnosis among patients referred for neurological evaluation to estimate their labour capacities at the unit of National Institute of Social Security (INSS), Florianopolis-SC, southern Brazil. METHOD: Review of all medical records of 108 patients evaluated between October 97 and May 98. The sample was submitted to judicious medico-legal assessment to define their final diagnosis. RESULTS: Neurological evaluation disclosed as the commonest disorders, in decreasing order of frequency: epilepsy, rheumatic diseases, psychiatric illnesses, neurological disorders related to chronic alcoholism, head trauma syndrome and cerebrovascular diseases. CONCLUSION: Neurological disorders may be responsible for important disability among workers in our society. However, the potential for social rehabilitation, often underestimated, must be considered. Moreover, diverse non-neurological conditions used to be sent for neurological evaluation. PMID- 11400029 TI - [Study of association between the ser-9-gly polymorphism of the D3 dopaminergic receptor and schizophrenia]. AB - Two molecular genetic studies were undertaken to investigate the association between a ser-9-gly polymorphism in the dopamine D3 receptor gene and schizophrenia. The first study analysed 141 schizophrenic patients and 189 matched controls. In addition, an haplotype relative risk study was performed using 35 trios (mother, father, affected offspring). No allelic or genotype association was found in both studies. We conclude that this D3 polymorphism is not a risk factor for schizophrenia in our sample. PMID- 11400030 TI - [Bayonet finger and the use of illicit drugs]. AB - Prevalence of "bayonet finger", a semiologic sign of attention deficit and hyperactivity disorder (ADHD), was determined in male illicit drug users, admitted in psychiatric hospital. A control group, paired in sex, age and social status, was built from the general population. The sign was observed en 165/345 (47.8%) drug users and 13/50 (26%) controls, the difference being statistically significant. Bearers of this sign had 2.61 odds ratio, 1.84 relative risk and 92.7% probability for being drug users. The observations suggest that, for males: (1) ADHD and related syndromes should be present in approximately half of illicit drug users and should represent a vulnerability factor for becoming drug user; (2) individuals presenting "bayonet finger", mainly adolescent, should deserve special attention for prevention of drug use. PMID- 11400031 TI - [Incidence of meningitis in patients 0-12 years old in the Instituto de Medicina Tropical de Manaus]. AB - The author analyzes the incidence of meningitis in patients with age ranging 0-12 years old treated in Instituto de Medicina Tropical de Manaus, during a period of two years. The illness is correlated with the etiological agent, treatment, follow-up, immediate and late complications, as well as the death by the illness. Comparison of results in this hospital experience with the literature allows a profile of the situation of meningitis in the State of Amazonas, Brazil. PMID- 11400032 TI - [Family dynamics of autistic children]. AB - We studied 15 families with autistics, 15 with Down's syndrome and 15 asymptomatic children. Patients' age ranged from 5 to 15 years-old. The parents of these three families' groups were appraised in regard to their family dynamics, to relate those symptoms to the functioning of an autistic family, in a comparative study. Details were provided of the families, the overall autistic features, the autistic's family, the family and the mental health, their limitations and difficulties throughout the vital cycle. An attempt was made to locate the factors that aid the family in hindering the healthy emotional development of its members. The field research was achieved by use of the instruments of the Family Dynamics Evaluation, (Carneiro, 1983). The data gathered were statistically compared. Considering the family population studied (n = 45), we found that the autistics' families and victims of Down's syndrome made it difficult to sustain the emotional health of group members. We conclude that the autistic's family dynamics caused difficulties to the emotional health of the group's members. PMID- 11400033 TI - [Neurological manifestations in the intoxication of infants by dimethicone combined with homatropine: report of 6 cases]. AB - OBJECTIVE: to alert that use of dimethicone plus homatropine in infants up to two months, may cause episodes of transitory extrapyramidal disturbances. METHOD: report 6 infants up to two months old, in daily use of dimethicone plus homatropine, that presented typical symptoms of the basal ganglia dysfunction, characterized by repeated crisis of short duration with tonic back shift of the head (opisthotonos), deviation of the eyes upward with looking fixed and terror expression, maintained in extending hypertonia posture of the 4 members and cry and/or guttural sounds emission. RESULTS: the extrapyramidal symptoms disappeared (and did not return) after interruption of dimethicone plus homatropine. Abnormalities were not verified in neurological examination, EEG and blood tests. CONCLUSIONS: the associated dimethicone/homatropine can produce in infants with less than 2 months, basal ganglia disturbances. The differentiation of these symptoms with the widespread epileptic crisis is essential, in order to avoid the erroneous use of antiepileptic drugs. PMID- 11400034 TI - Epileptic manifestations induced by midazolam in the neonatal period. AB - Antiepileptic drugs may cause worsening of epilepsy by aggravating pre-existing seizures or by triggering new seizure types. There are several reports of adverse effects related to midazolam, but only a few authors reported epileptic manifestations. We report four newborns seen at the Neonatal Intensive Care Unit of our University Hospital, who developed seizures a few seconds after the administration of midazolam. It is difficult to identify the patients at risk, but it is important to be aware and recognize this situation. PMID- 11400035 TI - [Schizencephaly: report of eleven cases]. AB - A retrospective study comparing clinical and computerized tomography (CT) in 11 patients diagnosed as having schizencephaly was conducted. Seven of these patients were girls and four boys. Six of them had tetraparesis, three hemiparesis and one no motor deficits. Six had epilepsy and ten developmental delay. On CT examinations, 7 patients were found as having bilateral clefts and four unilateral defect. Eight had opened lip clefts and four had a closed lip defect. The commonest associated anomaly was an absent septum pellucidum (n=9), followed by subependymal nodules (n=4), hydrocephalus (n=2) and microcephaly (n=1). Despite magnetic resonance image is the gold-standard to diagnose neuronal migration anomalies, CT can be useful in showing typical aspects of schizencephaly. PMID- 11400036 TI - Neuro-Behcet: report of three clinically distinct cases. AB - We report three patients who collectively have very representative clinical forms of neuro-Behcet and different neurological findings. The first case, male, 49 years old, presents symptoms similar to multiple sclerosis. The second case, male 15 years old, presents with parenchymatous compromise and an association with antiphospholipid antibody. And the third case, female 25 years old, presents an acute meningitis. Neuro-Behcet must always be included as a differential diagnosis of neurological disorders that have any difficulties in establishing a definite diagnosis. PMID- 11400037 TI - Late onset temporal lobe epilepsy with MRI evidence of mesial temporal sclerosis following acute neurocysticercosis: case report. AB - The objective of this case report is to describe magnetic resonance imaging (MRI) evidence of mesial temporal sclerosis (MTS) in a patient with new onset temporal lobe epilepsy (TLE) and acute neurocysticercosis with multiple cysts. A 56 years old man with new onset headache, Simple Partial Seizures and Complex Partial Seizures underwent CT scan and lumbar puncture as diagnose proceeding. Multiple cysts and meningitis were identified, with a positive immunology for cysticercosis. Seizures were recorded over the left temporal region in a routine EEG. Treatment with albendazole was performed for 21 days, with clinical improvement and seizure remission after 4 months. An MRI scan 11 months after treatment, showed complete resolution of those cystic lesions and a left hippocampal atrophy (HA) with hyperintense T2 signal. The presence of HA and hyperintense T2 signal in this patient has not, to date, been associated with a poor seizure control. CONCLUSIONS: This patient presented with MRI evidence of left MTS after new onset partial seizures of left temporal lobe origin. Although we did not have a previous MRI scan, it is likely that this hippocampal abnormality was due to the acute inflammatory response to cysticercosis associated to repeated partial seizures. This suggests that acute neurocysticercosis associated with repeated seizures may cause MTS and late onset TLE. PMID- 11400038 TI - Congenital contractural arachnodactyly with neurogenic muscular atrophy: case report. AB - We report the case of a 3-(1/2)-year-old girl with hypotonia, multiple joint contractures, hip luxation, arachnodactyly, adducted thumbs, dolichostenomelia, and abnormal external ears suggesting the diagnosis of congenital contractural arachnodactyly (CCA). The serum muscle enzymes were normal and the needle electromyography showed active and chronic denervation. The muscle biopsy demonstrated active and chronic denervation compatible with spinal muscular atrophy. Analysis of exons 7 and 8 of survival motor neuron gene through polymerase chain reaction did not show deletions. Neurogenic muscular atrophy is a new abnormality associated with CCA, suggesting that CCA is clinically heterogeneous. PMID- 11400039 TI - Cerebrospinal fluid fistula as the presenting manifestation of pituitary adenoma: case report with a 4-year follow-up. AB - We report the case of a young woman who presented with cerebrospinal fluid (CSF) rhinorrhea due to an undiagnosed and untreated pituitary adenoma. The tumor had extended well beyond sella turcica and caused bony erosion. The patient initially refused surgery and was treated with bromocriptine and a radiation therapy. CSF leakage did not improved and she was submitted to surgery by the transsphenoidal approach with removal of a tumor mass located in sphenoid sinus and sellar region. Origin of the leak was localized and repaired with fascia lata and a lumbar subarachnoid drain was left in place for 5 days. After 4 years she has normal serum PRL levels and no rhinorrhea. The management, complications and mechanisms involved in this rare condition are discussed. PMID- 11400040 TI - [Neurosarcoidosis: case report]. AB - We report a case of sarcoidosis in a 21 years old male with history of headache since he was three years old, with absence of abnormalities in a brain computerized tomography (CT). Brain magnetic resonance (MRI) disclosed a meningeal involvement with intense and homogeneous enhancement of the right fronto-temporal convexity and anterior third of the interhemispheric fissure. The brain MRI is more sensitive than brain CT and is the most appropriate method for the evaluation of meningeal and parenchymatous lesions, as well as monitoring the response to therapy and patients follow up. PMID- 11400041 TI - [Vasculitic neuropathy in Wegener granulomatosis: case report]. AB - The neurologic involvement during Wegener granulomatosis reaches 15-50% of patients, however as initial symptomatology is not commented in the literature. We describe a patient with Wegener's granulomatosis and mononeuritis multiplex, emphasizing the fact that neurologic manifestations anticipated systemic symptoms, and also focusing on the diagnostic contribution of sural biopsy and the good outcome of neurologic disease. PMID- 11400042 TI - [Transverse myelopathy in an adult with acute lymphoblastic leukemia: case report]. AB - We report a case of transverse myelopathy in a 31 year old white man with acute lymphoblastic leukemia, subtype L3 (ALL-L3). This is a severe form of leukemia that affects children more often than adults. Less than 1% of leukemic patients develop neurologic complication in the spinal cord. The symptomatology in the present case started with back pain, flaccid paraplegia, and loss of sensibility and vegetative functions below the lesion. The etiologic diagnostic was obtained through peripheral blood study, bone marrow cytology, cerebrospinal fluid analysis and magnetic resonance image of the dorsal cord. The antileukemic treatment with specific drugs had no influence on the fatal outcome of the disease. PMID- 11400043 TI - [Carotid-cavernous fistula with lethal epistaxis: case report]. AB - We report the case of a 32 years old male patient with carotid-cavernous fistula caused by head injury who died from massive epistaxis. Treatment assessment of this case is faced with the literature. PMID- 11400044 TI - [Trans-sphenoidal meningoencephaloventriculocele in an asymptomatic adult: case report]. AB - We report a casual finding of a sphenoethmoidal meningoencephaloventriculocele in an adult patient that the tomography finding was misinterpreted as an expansive neoplastic lesion with malignant features. We alert for the rare site of localization of this malformative pathology. PMID- 11400045 TI - [Parkinsonian, cerebellar, psychotic and demential symptoms in ex-boxer: case report]. AB - 67 years-old male patient, ex-boxer, after the end of his sportive career, began to present neuropsychiatric manifestations possibly due to the successive brain injuries related to the boxing. In report to this case, we discuss the mean organic psychosyndromes related to the boxing. PMID- 11400046 TI - [Sphenoidotomy approach to the sellar tumors: technical note]. AB - Dissection of mucosa from the nasal septum during transphenoidal approach can lead to significant morbidity. We report our experience with sphenoidotomy approach to the sella. This procedure obviates this dissection and its complication as septal perfurations, anosmy and epistaxis. Sphenoidotomy can be done under the microscopic view, endoscopic techniques and even with headlight. This approach is a safe and effective alternative to traditional or endoscopic exposures to the sella. PMID- 11400047 TI - HTLV-I negative tropical spastic paraparesis: a scientific challenge. AB - We reviewed the historical, clinical and etiological aspects of the progressive chronic spastic myelopathies of unknown etiology, disserting on the clinical similarities between HTLV-I seropositive and seronegative tropical spastic paraparesis (TSP), as well as focusing on the PCR studies of the seronegative TSP. PMID- 11400048 TI - Charcot and Brazil. AB - OBJECTIVE: To describe the relationship between Professor Charcot and Brazil. BACKGROUND: During the XIX century, French Neurology and its most prominent figure, Professor Charcot, dominated the area of nervous system diseases in the world. METHOD: We have reviewed some of the main publications about Charcot's life, the biography of Dom Pedro II, Emperor of Brazil and the development of Neurology in Brazil. RESULTS: Among the most important patients in Charcot's practice was the Emperor of Brazil. Dom Pedro II became a close friend of Charcot and he was a distinguished guest at Charcot's house, particularly at Tuesday soirees on boulevard St. Germain. In 1887, during the visit of Dom Pedro II to France, Charcot evaluated him and made the diagnosis of surmenage. In 1889, Dom Pedro II was disposed and went to Paris, where he lived until his death in 1891. Charcot signed the death certificate and gave the diagnosis of pneumonitis. Charcot had a passionate affection for animals, a feeling shared by Dom Pedro II. Dom Pedro II was affiliated to the French Society for the Protection of Animals. It is conceivable that Charcot's little monkey, from South America, was given to him by Dom Pedro II. The Brazilian Neurological School was founded by Professor A. Austregesilo in 1911, in Rio de Janeiro. At the time, of Charcot's death in 1893, his influence was still very important in the whole world. He and his pupils played a major role in the development of Brazilian Neurology. CONCLUSION: Professor Charcot had a close relationship with the Emperor of Brazil, Dom Pedro II. He was his private physician and they were close friends. The neurological school, created by professor Charcot, contributed significantly, albeit in an indirect way, to the development of Brazilian Neurology, starting in 1911, in Rio de Janeiro, by Professor A. Austregesilo. PMID- 11400049 TI - The effect of culture media on antigenic expression in sulfate-reducing bacteria. AB - The importance of sulfate-reducing bacteria (SRB) in nature has been widely recognized for many years. However, little is known about the ecology of SRB. The problem has been detecting, classifying, and quantifying these organisms. There are many shortcomings in the use of culture media for this purpose. As an alternative, fluorescent antibody (FA) techniques were considered as a method for the detection and identification of SRB. Antisera were prepared against whole cells of different species of SRB and evaluated for detection and identification of these organisms. Surface antigens of SRB were species specific. In addition, culture conditions influenced the expression of surface antigens, causing the antisera to be extremely specific. These results were confirmed by the sodium dodecyl sulfate-polyacrylamide-gel electrophoresis (SDS-PAGE) profiles of membrane proteins. On the basis of this specificity, the application of FA produced against culture collection strains would have limited application for detecting, identifying, and enumerating these organisms in nature. PMID- 11400050 TI - Periplasmic PQQ-dependent glucose oxidation in free-living and symbiotic rhizobia. AB - The expression of the pyrroloquinoline quinone (PQQ)-dependent glucose dehydrogenase (GDH) of Rhizobium tropici CIAT899 and Sinorhizobium meliloti RCR2011 was investigated under different nutrient-limiting conditions in continuous cultures, under different conditions of phosphate availability, and in S. meliloti bacteroids. The presence of free PQQ in alfalfa root exudates has also been assayed. It was shown that apo-GDH or holoenzyme was actively synthesized by these rhizobia, with the concomitant production of gluconate from glucose, under certain environmental conditions. GDH activity was also detected in bacteroids from alfalfa root nodules inoculated with either S. meliloti RCR2011 or 102F34. It was also shown that free PQQ was present in root exudates of alfalfa, but its production is ascribed to the activity of Erwinia sp., a normal contaminant of these seeds. PMID- 11400051 TI - Improved method for detection of methanotrophic bacteria in forest soils by PCR. AB - A primer set was designed for the specific detection of methanotrophic bacteria in forest soils by PCR. The primer sequences were derived from highly conservative regions of the pmoA gene, encoding the alpha-subunit of the particulate methane monooxygenase present in all methanotrophs. In control experiments with genomic DNA from a collection of different type I, II, and X methanotrophs, it could be demonstrated that the new primers were specific for members of the genera Methylosinus, Methylocystis, Methylomonas, Methylobacter, and Methylococcus. To test the suitability of the new primers for the detection of particulate methane monooxygenase (pMMO) containing methanotrophs in environmental samples we used DNA extracts from an acid spruce forest soil. For simple and rapid purification of the DNA extracts, the samples were separated by electrophoresis on a low-melting-point agarose gel. This allowed us to efficiently separate the DNA from coextracted humic acids. The DNA from the melted agarose gel was ready for use in PCR reactions. In PCR reactions with DNA from the Ah soil layer, products of the correct size were amplified by PCR by use of the new primers. By sequencing of cloned PCR products, it could be confirmed that the PCR products represented partial sequences with strong similarity to the pmoA gene. The sequence was most related to the pmoA sequence of a type II methanotroph strain isolated from the Ah layer of the investigated soils. PMID- 11400052 TI - Phosphorylation of LHI beta during membrane synthesis in the photosynthetic bacterium Rhodovulum sulfidophilum. AB - Cells of Rhv. sulfidophilum were grown under different conditions in the presence of 32P-phosphate and the corresponding H and L membrane fractions obtained and fractionated by SDS-PAGE. Both membranes showed almost identical polypeptide composition. The bacteriochlorophyll (Bchl) specific content in H was always lower that in L. As described before, oxygen did not regulate gene expression. Under high light, an almost two- to threefold decrease of the cellular specific Bchl content was observed. Pulse and chase experiments showed that transitions from aerobiosis to light-anaerobiosis did not quantitatively affect the Bchl content of the membranes, although a turnover of the 32P-phosphate and 35S methionine was observed. LHI beta was the only polypeptidic subunit of the Bchl binding polypeptides that was phosphorylated in vivo, and phosphotyrosine was the only phosphorylated amino acid detectable. The phosphorylated LHI beta was determined to be insoluble in the organic solvent mixture of (vol/vol) 1:1 chloroform-methanol containing ammonium acetate (0.1 m final concentration). Treatment with a chaotropic agent such as Na2CO3 solubilized the phosphorylated LHI beta, indicating that part of this posttranslationally modified polypeptide was not inserted in a transmembrane position. These results were used to speculate about the regulatory properties of this posttranslational modification of LHI beta on membrane differentiation. PMID- 11400053 TI - Sensitivity of partially purified ice nucleation activity of Fusarium acuminatum SRSF 616. AB - Factors that affect bacterial ice nucleation, including growth medium, growth phase, nutrient deprivation, and cold-temperature exposure, were investigated in the ice nucleation active (INA) fungus Fusarium acuminatum SRSF 616. Ice nucleation activity remained relatively constant throughout the growth cycle, and the cell-free culture supernatant consistently displayed higher ice nucleation activity than the hyphal pellet. Although nutrient starvation and low-temperature exposure enhance bacterial ice nucleation activity, reducing the concentration of C, N, or P in synthetischer nahrstoffarmer broth (SNB) did not increase fungal ice nucleation activity, nor did exposure to 4 degrees C or 15 degrees C. From the SNB supernatant, selected INA chromatography fractions were obtained that demonstrated increased sensitivity to proteinase K and heat compared with culture supernatant. We propose that partial purification of the fungal ice nuclei resulted in removal of low-molecular-weight stabilizing factors. PMID- 11400054 TI - Fluorescein diacetate hydrolysis as a measure of fungal biomass in soil. AB - The fatty acid methyl esters of lipids extracted from an agricultural soil in the preharvest period of soybean or middle growth cycle from wheat were characterized and quantified by gas-liquid chromatography. The fatty acids 18:2omega6 and 16:1omega5 were used as markers of saprotrophic and arbuscular mycorrhizal fungi. In parallel, biomass estimation through plate counts in selective media for cellulolytic and saprotrophic fungi was also performed all throughout a soybean crop or middle growth cycle of wheat. As an enzymatic method, the fluorescein diacetate (FDA) hydrolytic activity of the samples was determined. Owing to the high relationship exhibited by FDA hydrolysis with organic carbon and total nitrogen content of soil, the enzymatic activity was correlated with the microbial biomass estimated through marker lipids or plate counts. The results obtained point out that FDA hydrolysis may be used as a rapid, cheap, and reliable estimator of fungal biomass. PMID- 11400055 TI - Phosphotransbutyrylase expression in Bacillus megaterium. AB - The molecular analysis of a genomic region of B. megaterium revealed the presence of a gene coding for the enzyme phosphotransbutyrylase (Ptb). The enzyme activity was measured throughout the different phases of growth in B. megaterium, and its activity was found to be maximal in the late exponential growth phase. The branched amino acids isoleucine and valine activated Ptb expression. PtbBm was capable of using butyryl-CoA and 2-methyl-propionyl CoA as substrates. ActBm, a final sigma54 regulator from B. megaterium whose gene is situated upstream from the ptb gene, activated its expression. PMID- 11400056 TI - Cloning and sequence analysis of the endopolygalacturonase gene from the pitch canker fungus, Fusarium circinatum. AB - The fungus Fusarium circinatum causes pitch canker disease on mature pine trees and root rot and damping-off of pine seedlings. Endopolygalacturonases (endoPGs) play a major role during penetration of plants by fungi. Digestion of the pectic polysaccharides in the plant primary cell walls is one of the earliest functions of endoPGs during infection. The research objective was to clone and characterize the gene encoding endopolygalacturonase in F. circinatum. A 970-bp DNA fragment was cloned by using degenerate PCR amplification from F. circinatum DNA. Sequence data for this fragment were used to design specific primers for use in genome walking to amplify and sequence the remaining portion of the F. circinatum endoPG gene (Fcpg). The amino acid sequence predicted from this gene showed 90% and 87% similarity to Fusarium oxysporum and Fusarium moniliforme endoPGs, respectively. PMID- 11400057 TI - Molecular evidence for a Fe-hydrogenase in the green alga Scenedesmus obliquus. AB - Fe-hydrogenase-specific degenerate primers were used in polymerase chain reactions with single-stranded Scenedesmus obliquus cDNA as template. A resulting 435-bp fragment was used to screen a cDNA library from S. obliquus. A 1.6-kb cDNA clone, containing the 3'-end of an open reading frame, was obtained. Its deduced amino acid sequence contains the conserved H-cluster motives unique for Fe hydrogenases. Any other FeS-clusters seem to be absent. Phylogenetically, the enzyme is closely related to the Trichomonas vaginalis Fe-hydrogenase. Northern blot analysis shows that the Fe-hydrogenase is constitutively expressed in S. obliquus. Southern blot analysis of plastidic, mitochondrial, and nuclear DNA reveals that the enzyme is a single-copy gene encoded in the nucleus. Effects of transcriptional and translational inhibitors on H2-metabolism reveal the involvement of a chloroplast-encoded protein. PMID- 11400058 TI - Characterization of vitronectin-binding proteins of Staphylococcus epidermidis. AB - Staphylococcus epidermidis is the most common microorganism that is isolated from the cerebrospinal fluid (CSF) shunt infection patients. Vitronectin adsorbed on the surface of implants may mediate bacterial adhesion and colonization. To characterize vitronectin-binding properties, we analyzed S. epidermidis BD5703 isolated from a CSF shunt infection. Expression of vitronectin-binding protein(s) depended on culture media. Two proteins (60 and 52 kDa) were purified from vitronectin affinity chromatography. Two other vitronectin-binding proteins (21 and 16 kDa) were purified from an ion-exchange column. All purified proteins blocked bacterial binding of immobilized vitronectin significantly except the 16 kDa protein. The N-terminal sequences of the 21- and 16-kDa proteins did not show any appreciable amino acid sequence homology. The 52-kDa protein was sequenced by mass spectrometry and identified as an autolysin. This report demonstrates that interaction of vitronectin with multiple recognition sites on BD5703 surface may contribute to bacterial colonization. PMID- 11400059 TI - Substrate specificity of the highly alkalophilic bacterial proteinase esperase: relation to the x-ray structure. AB - Esperase is a highly alkalophilic bacterial proteinase produced by Bacillus lentus. The enzyme hydrolyzes peptide bonds comprising the carboxylic groups of hydrophobic as well as hydrophilic residues in the oxidized insulin B chain. Some of these bonds are not attacked by other alkaline microbial proteinases. P1-P4 specificity was determined by a series of peptide nitroanilides. The S1 recognition loop exhibits a preference for Phe. The "cleft" of the smallest subsite S2 prefers Ala and exhibits low affinity for the larger chain of Leu. S3 is more open than the other subsites and can accept a variety of residues. Hydrophobic interactions predominate in the S4-P4 interactions because S4 can accommodate Phe very well. The results characterize Esperase as an endopeptidase with a broader specificity in comparison with other microbial serine proteinases. This is probably owing to a more flexible substrate binding site. PMID- 11400060 TI - The corynebacterial flora of the normal human conjunctiva is lipophilic. AB - Corynebacterial isolates, obtained by selective and non-selective media from conjunctival swabs, were speciated by using two recently developed systems. All 25 strains from 92 individuals, as well as 12 strains from a previous similar study, belonged to lipophilic species. The significance of this unusual finding is discussed. PMID- 11400061 TI - Enhanced UV sensitivity of Thiobacillus ferrooxidans resulting from caffeine and acriflavine treatment of irradiated cells. AB - This study was aimed at identifying the roles of caffeine and acriflavine, two repair inhibitors, on UV sensitivity of iron-oxidizing Thiobacillus ferrooxidans ATCC 13728. The UV-dose response survival curve was inflected in nature, suggesting the population heterogeneity of the isolate. Caffeine and acriflavine potentiated the UV-induced killing of the organism. With the increase in concentrations of these compounds, the extent of survival decreased. Similarly, the inhibitory effects of caffeine and acriflavine increased with the increase in dose of UV-irradiation. The cells irradiated with 10 s (equivalent to 5.6 x 10( 5) J/m2/s) of UV-exposure tended to become resistant to the inhibitory effects of caffeine and acriflavine, as evidenced by the time course study of recovery. The cells appear to stage a dramatic recovery from UV damage in the presence of caffeine (3.0 mg/ml) and acriflavine (20 microg/ml) over a period of 25-30 h and 35-40 h respectively, when grown in the presence of energy sources. PMID- 11400062 TI - Assimilation of D-malate by Rhodobacter capsulatus E1F1. AB - Rhodobacter capsulatus grew by using either L- or D-malate as carbon sources under light/anaerobic conditions. The cellular yields were the same with D- or L malate. Both L-malate dehydrogenase and L-malic enzyme activities were detected in cell-free extracts from cells grown in both isomers. By contrast, a racemase activity converting D-malate into L-malate was induced only when D-malate was present in the culture medium. This racemase activity was Mn2+-dependent and was measured by coupling it either to the malate dehydrogenase or to the fumarase activities. The racemase activity was partially purified by anion-exchange chromatography. PMID- 11400063 TI - Construction of a shuttle vector and transformation of Xylella fastidiosa with plasmid DNA. AB - We have isolated, cloned, and sequenced a 5823-bp cryptic plasmid from a strain of Xylella fastidiosa. This plasmid encodes five open reading frames (ORF) greater than 400 nucleotides each. ORF 2 encodes a protein with 37% amino acid identity to the replication initiator protein of plasmid pECB2 from Pseudomonas alcaligenes. This RepA protein from X. fastidiosa contains both a leucine zipper and helix turn helix motif characteristic of proteins involved in DNA replication. The sequence 5' of ORF 2 has all of the features characteristic of plasmid origins of replication as well as regulatory elements required for transcription of ORF 2. Open reading frame 2, along with the upstream origin of replication, was cloned as an EcoRI fragment into pUC19 to create a shuttle vector. This construct was introduced into Xylella fastidiosa by electroporation, with selection for carbenicillin resistance. Transformation was verified by both PCR and Southern hybridization experiments. Frequency of transformation was low, but increased ten-fold when the plasmid was grown in X. fastidiosa rather than Escherichia coli prior to transformation. This work represents the first step towards the development of a system for genetic analysis of this important plant pathogen of citrus, grapevines, and other horticultural crops. PMID- 11400064 TI - Effects of mycoplasmal LAMPs on receptor responses to steroid hormones in mammalian cells. AB - Many individuals are chronically infected or parasitically colonized with mycoplasmas in their respiratory or urogenital tracts without apparent clinical significance. However, prolonged close interaction between prokaryotic agents and eukaryotic host cells may gradually and significantly alter normal biological or physiological properties of infected hosts. Steroid hormones are associated with rates of cancer formation in human. The purpose of this study is to establish a sensitive reporting system to examine whether mycoplasmal infections affect biological responses to steroid hormones in mammalian cells. We established pMTV CAT stably transfected cell lines to test the effect of mycoplasmal lipid associated membrane proteins (LAMPs). Results showed that LAMPs (1 microg/ml) from seven different species of human mycoplasmas-M. penetrans, M. fermentans, M. genitalium, M. salivarium, M. pneumoniae, M. orale, and M. hominis-had an inhibitory effect on androgen receptor (AR) response to 5alpha dihydrotestosterone (DHT) in the E82 transfectants. The inhibitory effect of mycoplasmal LAMPs appeared to be dose dependent. LAMPs from M. penetrans, M. genitalium, M. salivarium, M. pneumoniae, and M. orale also had an inhibitory effect on glucocorticoid receptor (GR) response to hormone dexamethasone (Dex) in TSU transfectants. In contrast, LAMPs from M. fermentans and M. hominis showed a stimulatory effect on the GR response to Dex in these TSU cells. The results suggest that colonization or chronic infection by mycoplasmas may significantly affect the responses of mammalian host cells to various steroid hormones, potentially affecting rates of cancer formation. PMID- 11400065 TI - The N-terminal signal sequence and the last 98 amino acids are not essential for the secretion of Bacillus sp. TS-23 alpha-amylase in Escherichia coli. AB - A truncated Bacillus sp. TS-23 alpha-amylase gene lacking 96 and 294 bp at its 5' and 3' end respectively was prepared by polymerase chain reaction and cloned into Escherichia coli expression vector, pQE-30, under the control of T5 promoter. SDS PAGE and activity staining analyses showed that the His6-tagged amylase had a molecular mass of approximately 54 kDa. Isopropyl-beta-d-thiogalactopyranoside (IPTG) induction of E. coli M15 cells bearing the recombinant plasmid resulted in the extracellular production of active amylase. Western blot analysis also revealed that the truncated amylase was present in the periplasmic space and culture medium. PMID- 11400066 TI - Adaptive and cross-protective responses of Pseudomonas sp. DJ-12 to several aromatics and other stress shocks. AB - Pseudomonas sp. DJ-12 cells were subjected to mild treatments of stress such as exposure to biphenyl, 4-chlorobiphenyl (4CB), 4-hydroxybenzoate (4HBA), ethanol, and heat, and then were examined for production of stress-shock proteins and morphological changes. The adapted cells were then subjected to lethal stress conditions such as 200 mm 4CB, 100 mm biphenyl, 10 mm 4HBA, 20% ethanol, and 46 degrees C to examine crossly protective responses to the stresses. Several stress shock proteins including DnaK and GroEL were newly synthesized in the adapted cells. Some of them were commonly produced by those stresses separately treated. The cells treated with these aromatic hydrocarbons showed destructive openings on the cell envelopes. On the other hand, those cells treated with ethanol or heat displayed irregular rod shapes with wrinkled surfaces. The adapted cells to each stress under sublethal conditions exhibited increased resistance to the same stress of lethal conditions. The cells adapted with 5 mm 4HBA showed greater protection for survival than those adapted by other stresses. In addition, those adapted cells showed increased resistance to other stresses as a cross-protection phenomenon. The cells adapted to 42 degrees C exhibited markedly increased resistance to the lethal stresses of 46 degrees C as well as to 20% ethanol. PMID- 11400067 TI - Cyanide production by rhizobacteria and potential for suppression of weed seedling growth. AB - Rhizobacteria strains were characterized for ability to synthesize hydrogen cyanide and for effects on seedling root growth of various plants. Approximately 32% of bacteria from a collection of over 2000 isolates were cyanogenic, evolving HCN from trace concentrations to > 30 nmoles/mg cellular protein. Cyanogenesis was predominantly associated with pseudomonads and was enhanced when glycine was provided in the culture medium. Concentrations of HCN produced by rhizobacteria were similar to exogenous concentrations inhibiting seedling growth in bioassays, suggesting that cyanogenesis by rhizobacteria in the rhizosphere can adversely affect plant growth. Growth inhibition of lettuce and barnyardgrass by volatile metabolites of the cyanogenic rhizobacteria confirmed that HCN was the major inhibitory compound produced. Our results suggest that HCN produced in the rhizospheres of seedlings by selected rhizobacteria is a potential and environmentally compatible mechanism for biological control of weeds. PMID- 11400068 TI - Desulfurization of benzothiophene by the Gram-negative bacterium, Sinorhizobium sp. KT55. AB - Sinorhizobium sp. KT55 was the first Gram-negative isolate to be capable of utilizing benzothiophene as the sole source of sulfur. By GC-MS analysis of metabolites of benzothiophene by this strain, benzothiophene sulfone, benzo[e][1,2]oxathiin S-oxide and o-hydroxystyrene were detected, suggesting that the benzothiophene desulfurization pathway of this strain is benzothiophene- >benzothiophene sulfoxide-->benzothiophene sulfone-->benzo[e][1,2]oxathiin S oxide-->o-hydroxystyrene. Desulfurization activity of this strain was significantly repressed by methionine, cysteine, sulfate, dimethyl sulfoxide, and Casamino acids. PMID- 11400069 TI - Cloning and nucleotide sequence of a new insertion sequence, IS231N, from a non serotypable strain of Bacillus thuringiensis. AB - A new IS231 variant, IS231N, has been isolated from an autoagglutinable, non serotypable strain of B. thuringiensis. IS231N is 1654 bp in length and is delimited by two incomplete 20-bp inverted repeats (IRL and IRR) with two mismatches. No direct repeats (DRs) were found at the right and left borders of IS231N. Surprisingly, IS231N contains three open reading frames (ORFs) that could code for polypeptides of 329 (ORF1), 118 (ORF2), and 17 (ORF3) amino acids, respectively. IS231N lacks the 5th conserved amino acid domain, called C2, owing to the addition of an adenine residue at nucleotide 1319. IS231N shows the highest nucleotide identity (99%) with IS231M, another insertion sequence previously isolated from the same bacterial strain. IS231N, however, shares only 83% amino acid identity with IS231M because of nucleotide substitutions and additions. The ORF1 of IS231N has five fewer amino acids than ORF1 of IS231M. Furthermore, the ORF2-3 putative fusion product in IS231N contains eight fewer amino acids than ORF2 in IS231M. The dendrogram showing the evolutionary relationship between members of the IS231 family and IS231N indicates that IS231N is phylogenetically more closely related to IS231M (83%), followed by IS231F(74%), and is more distant from IS231V and W(46%). PMID- 11400070 TI - Production of a novel compound, 7,10,12-trihydroxy-8(E)-octadecenoic acid from ricinoleic acid by Pseudomonas aeruginosa PR3. AB - The production and its potential use of a novel trihydroxy unsaturated fatty acid, 7,10,12-trihydroxy-8(E)-octadecenoic acid (TOD), were investigated. TOD was formed by Pseudomonas aeruginosa PR3 (NRRL B-18602) in a culture supplied with exogenous ricinoleic acid. The yield of TOD production was always higher in a rich culture medium than in minimal screening medium. Extending the conversion time from 48 to 72 h prior to lipid extraction led to a 65% reduction in yield, indicating that TOD was further metabolized by strain PR3 and that control of reaction time is important to achieving a maximum yield. The optimum culture density, reaction time, pH, temperature, and substrate concentration for the production of TOD were: 20-24 h culture growth, 48 h, 7.0, 25 degrees C, and 1% (vol/vol), respectively. Under optimum conditions, the yield of TOD production was greater than 45%. TOD was found to be an antifungal agent most active against the fungus that causes blast disease in rice plants, the most important fungal disease affecting rice production worldwide. PMID- 11400071 TI - Acanthamoeba can be differentiated by the polymerase chain reaction and simple plating assays. AB - Acanthamoeba are opportunistic pathogens with invasive and noninvasive species. For clinical purposes it is important to differentiate potentially pathogenic from nonpathogenic isolates. For the rapid and sensitive identification of Acanthamoeba at the genus level, we used a polymerase chain reaction (PCR)-based method which detected as few as five cells. Further, we tested nine isolates of Acanthamoeba for their ability to produce cytopathic effects (CPE) on corneal epithelial cells. On the basis of the results, Acanthamoeba were divided into pathogenic or nonpathogenic groups. However, because CPE assays are not available to every diagnostic laboratory, we developed a simple plating assay based on osmotolerance which correlated well with the CPE assays. Pathogenic Acanthamoeba showed growth on higher osmolarity (agar plates containing one molar mannitol), while growth of nonpathogens was inhibited on these plates. In conclusion, we have developed methods for the rapid identification and differentiation of Acanthamoeba. PMID- 11400072 TI - Potential intervention of Campylobacter jejuni in the modulation of murine immune response. AB - Campylobacter jejuni has been reported to produce different toxins that may modulate the immune response in both animals and humans. The effect of C. jejuni enterotoxin on the immune response was investigated in two groups of Balb/c mice. One of them was inoculated intraperitoneally with 1010 colony forming units (CFU) of an enterotoxigenic strain (CCUG 7580), and the second one with a non enterotoxigenic strain (CCUG 7440). The number of polymorphonuclear (PMN) cells from spleen increased in both enterotoxigenic and non-enterotoxigenic strains as a consequence of C. jejuni infection. Notwithstanding, lymphocyte proliferation stimulated by lipopolysaccharide (LPS) was increased by both enterotoxigenic and non-enterotoxigenic strains. Interleukin-2 (IL-2) production from splenic cells was increased significantly by infection with the enterotoxigenic strain. Both enterotoxigenic and non-enterotoxigenic strains reduced the splenic response to sheep erythrocytes; the response was significantly suppressed for immunoglobulin M (Ig M) and for immunoglobulin G (Ig G) synthesis. These results suggest that C. jejuni is able to modify some components of the immune response in mice, and also that the enterotoxigenic strain has more immunomodulating activity than the non enterotoxigenic strain. PMID- 11400073 TI - Differences in Escherichia coli culture conditions can have a large impact on the induction of extreme acid resistance. AB - A recently isolated Escherichia coli strain (3TF4) survived an acid shock that mimicked the low pH of the human gastric stomach (pH 2, 1 h), but this survival was highly influenced by prior growth conditions. Only 0.01% of the stationary phase cells that had been grown anaerobically in a carbonate medium (2 mg glucose and 0.25 mg yeast extract per ml, 40 mm sodium carbonate, final pH 6.5) survived the acid shock, and the survival of exponential phase cultures was even lower (0.0001%). Small amounts of Trypticase (1.5 mg/ml) increased the survival as much as 5000-fold, but cultures that were provided with higher concentrations of Trypticase (7.5 mg/ml) did not reach the stationary phase in 24 h and were more acid sensitive. Sodium acetate (50 mm) also increased acid resistance, and the increased acid shock survival was greater for the cells that had reached the stationary phase (100 versus 1000-fold, respectively). E. coli 3TF4 cultures that had been grown aerobically in Luria broth were already so acid resistant (survivals greater than 40%) that they did not respond to sodium acetate. E. coli 3TF4 cultures that were refrigerated (5 degrees C, 7 days) were nearly as acid resistant as those that were immediately subjected to acid shock (pH 2.0, 1 h). PMID- 11400074 TI - The effect of various carbonate sources on the survival of Escherichia coli in dairy cattle manure. AB - Manure slurries (n = 3) prepared from the feces and urine of lactating dairy cattle (1 part urine, 2.2 parts feces, and 6.8 parts distilled water) had an initial pH of 8.6 +/- 0.1; dissolved carbonate concentrations of 48 +/- 4 mm, and Escherichia coli counts of 5.9 +/- 0.7 logs per ml slurry. The pH of untreated slurries declined to pH 7.0 +/- 0.1 by the 10th day of incubation, and the E. coli count increased approximately 10-fold (P < 0.05). When slurries were treated with Na2CO3, K2CO3, NaHCO3 or Na2CO3.NaHCO3 (0 to 16 g/kg slurry), the dissolved carbonates increased in a linear fashion, but only Na2CO3 and K2CO3 (8 g/kg or greater) or Na2CO3.NaHCO3 (16 g/kg) ensured an alkaline pH. Even relatively low concentrations of Na2CO3 or K2CO3 (8 or 12 g/kg) caused a decrease in E. coli viability (P < 0.05), and E. coli could not be detected if 16 g/kg was added (day 5 or 10 of incubation). Na2CO3.NaHCO3 also caused a decrease in E. coli viability, (P < 0.05), but some E. coli (approximately 104 cells per g) were detected on day 10 even if the concentration was 16 g/kg. NaHCO3 did not prevent the decrease in pH or cause a decrease in E. coli numbers (P > 0.05). Calculations based on the Henderson-Hasselbalch equation (pH and dissolved carbonates) indicated that little E. coli killing was noted until the dissolved carbonate anion concentrations (CO3-2) were greater than 1 mm, but bicarbonate anion (HCO3-) concentrations as high as 180 mm did not affect E. coli viability. These results are consistent with the idea that carbonate anion has antimicrobial properties and can kill E. coli in dairy cattle manure. PMID- 11400076 TI - Absence of clonality of Campylobacter jejuni in serotypes other than HS:19 associated with Guillain-Barre syndrome and gastroenteritis. AB - Guillain-Barre syndrome (GBS) is recognized as a complication that occurs after Campylobacter infection. Certain Penner serotypes, such as HS:19, are linked particularly to GBS in some parts of the world, and there is good evidence for restricted genetic diversity in these isolates. However, GBS also occurs after Campylobacter infection due to other serotypes. Therefore, we asked whether Campylobacter jejuni non-HS:19 serotypes associated with GBS have a clonal structure and differ from strains isolated from patients with Campylobacter gastroenteritis. A worldwide selected population of C. jejuni non-HS:19 strains associated with GBS and gastroenteritis was analyzed by use of multilocus enzyme electrophoresis, automated ribotyping, pulsed-field gel electrophoresis, and flagellin gene typing. The results show that these isolates represent a heterogenic population and do not constitute a unique population across serotypes. No epidemiologic marker for GBS-associated strains was identified. PMID- 11400077 TI - Molecular population genetic analysis of Campylobacter jejuni HS:19 associated with Guillain-Barre syndrome and gastroenteritis. AB - Infection with Campylobacter jejuni serotype HS:19 is associated with the development of Guillain-Barre syndrome (GBS). To determine whether a particular HS:19 clone is associated with GBS, multilocus enzyme electrophoresis (MLEE) was used to analyze a worldwide collection of isolates. There were 34 electropherotypes (ETs) in 3 phylogenetic clusters among 83 C. jejuni isolates. Cluster I contained all HS:19 strains, and a single ET (ET4) accounted for most HS:19 strains. HS:19 strains did not occur in any of the other clusters. ET4 contained isolates from different geographic locations, indicating global spread of this clone. Furthermore, ET4 contained isolates from patients with uncomplicated enteritis and GBS, as well as isolates from animal sources. The results of this study show that HS:19 strains comprise a clonal, although not monomorphic, population, which is distinct from non-HS:19 strains within C. jejuni. A unique clone associated with GBS was not identified by use of MLEE. PMID- 11400082 TI - Hypercoagulable syndromes: evaluation and management strategies for acute limb ischemia. AB - Acute limb ischemia secondary to peripheral arterial thrombosis is a relatively uncommon but ominous form of vascular accident. Select inherited and acquired hypercoagulable states appear to contribute to an initial arterial thrombosis and, more importantly, recurrent thrombotic events. Mounting interest in hypercoagulability, the increased availability of hypercoagulable state "profiles," and enhanced ability to identify an abnormality in tested patients have promoted widespread testing. Unfortunately, widespread testing has had a limited beneficial impact on the management of acute limb ischemia. Ideally, costly and specialized testing should be limited to situations in which the results will have a tangible impact on patient care. Clear goals of testing should be determined before testing is performed. This article addresses a practical approach to hypercoagulable state testing in patients with acute limb ischemia with a focus on abnormalities that impact patient management. PMID- 11400083 TI - Demography and etiology of acute leg ischemia. AB - Acute leg ischemia arguably is the most challenging condition that a vascular surgeon has to manage. Patients have widely different presentations, there are several available treatment options, and, yet, despite best efforts, the outcome remains poor. An understanding of the changes in population demographics in affluent western societies, set in an historical context, enables choices about therapy. A vascular unit serving a community including half a million people can expect to treat approximately 75 patients with acute leg ischemia per year. It is vital that a team approach is used to decide management because endovascular treatment with peripheral thrombolysis is increasingly important. The distinction between etiology of thrombosis or embolism remains important, although the diagnosis can never be conclusive. The severity of arterial ischemia is a more important determinant of outcome; optimal management, therefore, begins with stratification according to the severity of ischemia on admission to the hospital. Subsequent care lends itself to organization by care-pathway or guidelines that take into account the available local facilities. PMID- 11400084 TI - Operation for acute peripheral arterial occlusion: is it still the gold standard? AB - Acute arterial ischemia secondary to peripheral arterial occlusion has been shown to cause severe morbidity and mortality. Debate continues about the best mode for initial therapy of patients presenting with acute limb ischemia (ALI). Surgery traditionally has been used as the sole mode of therapy. Since the introduction of catheter-directed thrombolysis (CDT), role of surgery as the "gold standard" has been questioned. In this report the authors review the role of surgery compared with CDT. They discuss the role of prompt diagnosis on the outcome of the intervention and the results of CDT compared with the surgical standard. The best therapy for ALI is the one that is instituted early; intervention should be tailored based on the initial clinical presentation, and surgery remains the gold standard with CDT, an adjunctive tool for the vascular surgeon dealing with acute peripheral arterial occlusion (PAO). PMID- 11400085 TI - Fluoroscopically assisted thromboembolectomy: should it be routine? AB - The treatment of acute limb ischemia secondary to thromboembolic disease has become significantly more difficult because of the increased proportion of elderly patients presenting with complex patterns of atherosclerotic disease. The presence of multiple medical comorbidities also complicates operative and perioperative management in these patients. Neither the techniques nor catheter designs have changed significantly since the introduction of the balloon embolectomy catheter in 1963, which permitted extraction of clot from remote sites. The authors believe that the use of intraoperative fluoroscopy and the performance of fluoroscopically assisted thromboembolectomy (FATE) greatly improves the results of this treatment in many cases. FATE facilitates catheter passage through tortuous, diseased arteries, identifies residual thrombus and underlying lesions, reduces vessel damage caused by balloon overinflation, and decreases the risk of catheter-induced dissection or atherosclerotic plaque displacement. Intraoperative fluoroscopy helps determine the need for as well as guides adjunctive procedures such as angioplasty and stenting. Such procedures can be performed at the time of the thromboembolectomy simplifying and expediting treatment. PMID- 11400086 TI - Reperfusion and compartment syndromes: strategies for prevention and treatment. AB - Reperfusion syndrome refers to the damage done by restoration of blood flow to ischemic tissues and is distinct from the original ischemic insult itself, whereas compartment syndrome refers to the damage resulting from increased pressure within an enclosed fascial compartment that occurs after blood flow has been restored. Despite extensive experimental work directed toward the treatment of established reperfusion injury and prevention of compartment syndrome, clinical outcome over the past decade has not appreciably changed. Although the systemic insult, thought to be an inevitable result of reperfusion injury, may be less injurious than "conventional wisdom" would suggest, no better strategy for treating compartment syndrome other than early recognition and decompression has yet been developed. PMID- 11400087 TI - Techniques for the percutaneous treatment of acute arterial occlusion. AB - There are many modalities utilized in the treatment of acute arterial occlusions. The purpose of this report will be to review the various studies defining percutaneous chemical thrombolysis, mechanical thrombectomy, and combination therapies. A clinical paradigm for acute limb ischemia will be defined, and a detailed description of techniques utilized in the above percutaneous interventions will be outlined. PMID- 11400088 TI - Mechanical thrombectomy as a first-line treatment for arterial occlusion. AB - The use of percutaneous mechanical thrombectomy devices to restore flow to an acutely ischemic limb is gaining popularity. Potential benefits include the minimally invasive nature of the procedure, rapid blood flow restoration, and a decrease in the dose and duration of adjunctive pharmacologic thrombolytic agents when required. A detailed description of the currently available mechanical thrombectomy devices, preclinical device evaluation, and published clinical trials for the management of acute limb-threatening ischemia are provided. PMID- 11400089 TI - Intraoperative lytic therapy: agents and methods of administration. AB - Intraoperative intraarterial thrombolysis is a valuable adjunct for the removal of residual arterial thrombi after mechanical thromboembolectomy. Early reports by some investigators indicated high rates of bleeding complications, most likely caused by inappropriately high doses of plasminogen activators infused over long periods of time, which led to systemic lytic effects. Animal models and controlled human experiments subsequently have shown the potential efficacy and safety of intraarterial thrombolysis. Since urokinase (UK) has been withdrawn from the market, recombinant tissue-type plasminogen activator (rt-PA) has become the plasminogen activator of choice. Reteplase and other plasminogen activators may be beneficial (and safe); however, data on intraoperative use currently are not available. The most common methods of delivery into the distal arterial tree are bolus infusion with inflow occlusion, drip infusion after restoration of arterial inflow, and the isolated limb perfusion technique. The number of distal vessels involved, the amount of residual thrombus, and severity of ischemia guide the dose of plasminogen activator, volume of perfusate, and the technique and duration of infusion. PMID- 11400090 TI - New strategies in platelet inhibition in noncoronary interventions. AB - Thrombembolic complications frequently occur during and after endovascular procedures because of associated arterial injury and thrombotic characteristics of implanted devices such as stents. New strategies in platelet aggregation inhibition are now available blocking the final and common pathway of platelet aggregation, the glycoprotein IIb/IIIa receptor. This treatment modality seems to be more effective for prophylaxis and prevention of thrombembolic complications than standard antiplatelet therapy. Most of the data provided for glycoprotein IIb/IIIa receptor blockade are derived from studies of coronary interventions. This report reviews the pharmacodynamic differences of classic and new drugs for platelet inhibition and the basic considerations for antiplatelet therapy in noncoronary interventions. PMID- 11400091 TI - Using efficacy, safety, and cost data to support a formulary decision regarding thrombolytic therapy. AB - When urokinase was withdrawn from the market, alternative thrombolytics such as alteplase and reteplase needed to be evaluated for peripheral vascular disease (PVD). The efficacy, safety, and cost of these agents were evaluated formally by the Department of Pharmacy and presented to the Pharmacy and Therapeutics Committee. No published data support a difference in efficacy or safety between these agents. A cost analysis estimated the average total cost of care was higher for patients treated with reteplase ($4,556) compared with alteplase ($2,139). Therefore, alteplase was determined to be a more cost-effective thrombolytic agent to treat PVD. PMID- 11400092 TI - Influence of anaerobic culture acclimation on the degradation kinetics of various substrates. AB - The adaptation of an anaerobic culture (anaerobic sludge) to a specific substrate brings significant changes to its microbial population. These changes can be described by the sludge's ability to treat various substrates such as carbohydrates or proteins or "intermediate" products of anaerobic metabolism such as L-lactic, propionic, and acetic acids. The activity of the sludge with respect to a specific substrate is a critical parameter, because the anaerobic degradability of wastewaters depends strongly on it. This work examines and quantifies the differentiation of two anaerobic sludges of the same origin, following an adaptation period of about 18 months to lactose and gelatin, respectively. The acclimation has a significant effect on the maximum specific utilization rates of various compounds and on their apparent consumption kinetics. It is noticeable, however, that even if the anaerobic cultures were not exposed to a specific substrate for a prolonged period of time (more than a year), they still kept the ability of hydrolyzing or degrading it. In addition, the acclimation has an unquestionable effect on the stoichiometry of the production of volatile fatty acids and L-lactate. Finally, from codigestion experiments it is shown that codigestion of lactose and gelatin appears to have no effect on their hydrolysis kinetics in any of the lactose or gelatin acclimated cultures; specifically, the hydrolysis kinetics remained the same as calculated when lactose or gelatin were the only fed substrates. Similarly, the kinetics of L-lactate and D-glucose biodegradation seemed to be unchanged. On the other hand, codigestion has a significant effect on the production of L-lactic, propionic, and acetic acids, which can be attributed to the increased hydrogen production accompanying gelatin biodegradation. PMID- 11400093 TI - Using isotopomer path tracing to quantify metabolic fluxes in pathway models containing reversible reactions. AB - As a more complete picture of the genetic and enzymatic composition of cells becomes available, there is a growing need to describe how cellular regulatory elements interact with the cellular environment to affect cell physiology. One means for describing intracellular regulatory mechanisms is concurrent measurement of multiple metabolic pathways and their interactions by metabolic flux analysis. Flux of carbon through a metabolic pathway responds to all cellular regulatory systems, including changes in enzyme and substrate concentrations, enzyme activation or inhibition, and ultimately genetic control. The extent to which metabolic flux analysis can describe cellular physiology depends on the number of pathways in the model and the quality of the data. Intracellular information is obtainable from isotopic tracer experiments, the most extensive being the determination of the isotopomer distribution, or specific labeling pattern, of intracellular metabolites. We present a rapid and novel solution method that determines the flux of carbon through complex pathway models using isotopomer data. This time-consuming problem was solved with the introduction of isotopomer path tracing, which drastically reduces the number of isotopomer variables to the number of isotopomers observed experimentally. We propose a partitioned solution method that takes advantage of the nearly linear relationship between fluxes and isotopomers. Whereas the stoichiometric matrix and the isotopomer matrix are invertible, simulated annealing and the Newton Raphson method are used for the nonlinear components. Reversible reactions are described by a new parameter, the association factor, which scales hyperbolically with the rate of metabolite exchange. Automating the solution method permits a variety of models to be compared, thus enhancing the accuracy of results. A simplified example that contains all of the complexities of a comprehensive pathway model is presented. PMID- 11400094 TI - Long-term performance of bioreactors cleaning mercury-contaminated wastewater and their response to temperature and mercury stress and mechanical perturbation. AB - The long-term performance of bioreactors retaining mercury from contaminated industrial wastewater was analyzed at the laboratory scale, and its response to mechanical perturbations (gas bubbles and shaking) as well as to physical (increased temperature and hydraulic load) and chemical stresses (increased mercury concentration) likely to occur during on site operation was studied. Two packed-bed bioreactors with 80-cm(3) lava chips as biofilm carrier were inoculated with nine Hg(II)-resistant natural isolates of alpha- and gamma proteobacteria. Chloralkali wastewater containing ionic mercury (3.0 to 9.7 mg/L Hg(2+)), amended with sucrose and yeast extract, flowed through the bioreactors at 160 mL/h. During the 16-month investigation the bioreactors showed no sign of depleted performance in terms of mercury-retaining capacity. After 16 months, both bioreactors still retained 96% of the mercury load. The performance of the bioreactors was sensitive to mechanical perturbations (e.g., sheer forces of gas bubbles). Shifts to higher Hg(2+) inflow concentrations initially decreased the mercury retention efficacy slightly. However, the bioreactors could adapt to Hg(2+) concentrations of up to 7.6 mg/L within several days. Old biofilms were less affected than the younger ones. The performance of the bioreactors was not affected by an increase in temperature up to 41 degrees C and an increased volumetric load (up to 240 mL/h). The bioreactors regained activity spontaneously after the stress had stopped. Recovery could be accelerated by increased nutrient concentration, although this may lead to blocking of the packed bed. PMID- 11400095 TI - An experimental and theoretical study of the inhibition of Escherichia coli lac operon gene expression by antigene oligonucleotides. AB - Previously, we have developed a genetically structured mathematical model to describe the inhibition of Escherichia coli lac operon gene expression by antigene oligos. Our model predicted that antigene oligos targeted to the operator region of the lac operon would have a significant inhibitory effect on beta-galactosidase production. In this investigation, the E. coli lac operon gene expression in the presence of antigene oligos was studied experimentally. A 21 mer oligo, which was designed to form a triplex with the operator, was found to be able to specifically inhibit beta-galactosidase production in a dose-dependent manner. In contrast to the 21-mer triplex-forming oligonucleotide (TFO), several control oligos showed no inhibitory effect. The ineffectiveness of the various control oligos, along with the fact that the 21-mer oligo has no homology sequence with lacZYA, and no mRNA is transcribed from the operator, suggests that the 21-mer oligo inhibits target gene expression by an antigene mechanism. To simulate the kinetics of lac operon gene expression in the presence of antigene oligos, a genetically structured kinetic model, which includes transport of oligo into the cell, growth of bacteria cells, and lac operon gene expression, was developed. Predictions of the kinetic model fit the experimental data quite well after adjustment of the value of the oligonucleotide transport rate constant (9.0 x 10(-)(3) min(-)(1)) and oligo binding affinity constant (1.05 x 10(6) M(-)(1)). Our values for these two adjusted parameters are in the range of reported literature values. PMID- 11400096 TI - Improved glycosylation of a foreign protein by Tn-5B1-4 cells engineered to express mammalian glycosyltransferases. AB - The major advantages of using the baculovirus-insect cell system for recombinant protein production are its ability to produce large amounts of recombinant proteins and its ability to provide eucaryotic modifications, such as glycosylation. However, the glycans linked to recombinant glycoproteins produced by this system typically differ from those found on native mammalian products. This is an important problem because glycans on mammalian glycoproteins can influence their functions in many different ways. The inability of baculovirus infected insect cells to produce glycans identical to those found on native mammalian glycoproteins is due, in part, to the absence of functional levels of certain glycosyltransferases in insect cells. Thus, the purpose of this study was to engineer these activities into Tn-5B1-4, an established insect cell line that is widely used as a host for baculovirus-mediated protein production. Expression plasmids were constructed in which cDNAs encoding mammalian beta1,4 galactosyltransferase and alpha2,6-sialyltransferase were placed under the transcriptional control of a baculovirus immediate early promoter. These plasmids were then used to isolate two different transgenic Tn-5B1-4 derivatives and the biological and biochemical properties of these cell lines were examined. The results show that both of the engineered insect cell lines have improved glycoprotein-processing capabilities, relative to the parental cell line. PMID- 11400097 TI - Efficient polycyclic aromatic hydrocarbons dihydroxylation in direct micellar systems. AB - Optimization of whole-cell bioconversion of the polycyclic aromatic hydrocarbons (PAHs) anthracene, phenanthrene, and naphthalene to the enantiomerically pure corresponding cis-dihydroxydihydro derivatives by the Escherichia coli JM109 (pPS1778) recombinant strain, carrying the naphthalene dioxygenase and corresponding regulatory genes cloned from Pseudomonas fluorescens N3, in micellar systems, is presented. We show that direct microemulsion systems, where a nonionic surfactant such as 1.5% (v/v) Triton X-100 plus 0.6% to 1.0% (v/v) selected oils are able to solubilize the PAHs tested at relatively high concentrations (initial concentrations in the reaction medium > or =10 mM for naphthalene and phenanthrene and > or =2 mM for anthracene), and allow for more efficient substrate bioconversion. These media, while not affecting bacteria viability and performance, provide increased efficiency and final product yields (100% for naphthalene, >30% for anthracene, >60% for phenanthrene). The phase behavior of the direct microemulsion systems for the different substrates and oils utilized was monitored as a function of their volume fraction by light scattering experiments, and related to the bioconversion results. For anthracene and phenanthrene, the dihydroxylated products have an inhibitory effect on the conversion reactions, thus hindering complete turnover of the substrates. We ascertain that such inhibition is reversible because removal of the products formed allowed the process to start over at rates comparable to initial rates. To allow for complete conversion of the PAHs tested a stepwise or continuous separation of the product formed from the micellar reaction environment is being developed. PMID- 11400098 TI - Enzyme stabilization by covalent binding in nanoporous sol-gel glass for nonaqueous biocatalysis. AB - A unique nanoporous sol-gel glass possessing a highly ordered porous structure (with a pore size of 153 A in diameter) was examined for use as a support material for enzyme immobilization. A model enzyme, alpha-chymotrypsin, was efficiently bound onto the glass via a bifunctional ligand, trimethoxysilylpropanal, with an active enzyme loading of 0.54 wt%. The glass bound chymotrypsin exhibited greatly enhanced stability both in aqueous solution and organic solvents. The half-life of the glass-bound alpha-chymotrypsin was >1000-fold higher than that of the native enzyme, as measured either in aqueous buffer or anhydrous methanol. The enhanced stability in methanol, which excludes the possibility of enzyme autolysis, particularly reflected that the covalent binding provides effective protection against enzyme inactivation caused by structural denaturation. In addition, the activity of the immobilized alpha chymotrypsin was also much higher than that of the native enzyme in various organic solvents. From these results, it appears that the glass-enzyme complex developed in the present work can be used as a high-performance biocatalyst for various chemical processing applications, particularly in organic media. Published by John Wiley & Sons PMID- 11400099 TI - A two-step enzymatic resolution process for large-scale production of (S)- and (R)-ethyl-3-hydroxybutyrate. AB - An efficient two-step enzymatic process for production of (R)- and (S)-ethyl-3 hydroxybutyrate (HEB), two important chiral intermediates for the pharmaceutical market, was developed and scaled-up to a multikilogram scale. Both enantiomers were obtained at 99% chemical purity and over 96% enantiomeric excess, with a total process yield of 73%. The first reaction involved a solvent-free acetylation of racemic HEB with vinylacetate for the production of (S)-HEB. In the second reaction, (R)-enriched ethyl-3-acetoxybutyrate (AEB) was subjected to alcoholysis with ethanol to derive optically pure (R)-HEB. Immobilized Candida antarctica lipase B (CALB) was employed in both stages, with high productivity and selectivity. The type of butyric acid ester influenced the enantioselectivity of the enzyme. Thus, extending the ester alkyl chain from ethyl to octyl resulted in a decrease in enantiomeric excess, whereas using bulky groups such as benzyl or t-butyl, improved the enantioselectivity of the enzyme. A stirred reactor was found unsuitable for large-scale production due to attrition of the enzyme particles and, therefore, a batchwise loop reactor system was used for bench scale production. The immobilized enzyme was confined to a column and the reactants were circulated through the enzyme bed until the targeted conversion was reached. The desired products were separated from the reaction mixture in each of the two stages by fractional distillation. The main features of the process are the exclusion of solvent (thus ensuring high process throughput), and the use of the same enzyme for both the acetylation and the alcoholysis steps. Kilogram quantities of (S)-HEB and (R)-HEB were effectively prepared using this unit, which can be easily scaled-up to produce industrial quantities. PMID- 11400100 TI - Immobilization of Candida rugosa lipase on colloidal gas aphrons (CGAs). AB - A novel technique for immobilization of Candida rugosa lipase onto anionic colloidal gas aphrons (CGAs) is described. CGAs are spherical microbubbles (10 100 microm) composed of an inner gas core surrounded by a surfactant shell. In this initial study, greater than 80% lipase (w/w) was effectively retained on the CGAs. Leakage of protein from the CGAs and the activity of the adsorbed lipase decreased with increasing enzyme loading; this indicates that multilayers of lipase may be adsorbing onto the CGAs. The CGA-immobilised lipase displayed normal Michaelis-Menten dependence on substrate concentration and also exhibited greater activity than the free enzyme. PMID- 11400101 TI - Rapamycin reduces hybridoma cell death and enhances monoclonal antibody production. AB - Rapamycin was used as a medium additive to slow the progression of CRL 1606 hybridomas through the cell cycle, under the hypothesis that such a modulation might reduce cell death. Cell cycle distributions for CRL hybridomas in the G1 phase of the cell cycle ranged from 20% to 35% during batch, fed-batch, and continuous culture experiments, independent of culture time, dilution rate, growth rates, or death rates. Rapamycin, an mTOR signaling inhibitor, immunosuppressant, and G1-phase arresting agent, was identified and tested for efficacy in restraining cell cycle progression in CRL 1606 hybridoma cultures. However, in the presence of 100 nM rapamycin, the percentage of cells in the G1 phase of the cell cycle during fed-batch cultures was only increased from 28% to 31% in control cultures to 37% to 48% for those with rapamycin. Accordingly, rapamycin only slightly reduced culture growth rate. Instead, the use of rapamycin more notably kept viability higher than that of control cultures by delaying cell death for 48 h, thereby enabling viable proliferation to higher maximum viable cell densities. Furthermore, rapamycin enhanced specific monoclonal antibody production by up to 100% during high-viability growth. Thus, over the course of 6-day fed-batch cultivations, the beneficial effects of rapamycin on viable cell density and specific productivity resulted in an increase in final monoclonal antibody titer from 0.25 to 0.56 g/L (124%). As rapamycin is reported to influence a much broader range of cellular functions than cell cycle alone, these findings are more illustrative of the influence that signal transduction pathways related to mTOR can have on overall cell physiology and culture productivity. PMID- 11400102 TI - Swimming characteristics of magnetic bacterium, Magnetospirillum sp. AMB-1, and implications as toxicity measurement. AB - To develop a novel toxicity measurement system using the persistent swimming property of magnetic bacteria along an externally applied magnetic field, certain characteristics of Magnetospirillum sp. AMB-1 cells were examined, including their growth pattern, motility, magnetosensitivity, swimming speed, and cell length distribution. In addition, the effect of toxic compounds on the swimming speed was assessed relative to application as a toxicity sensor. With an inoculum of 1.0 x 10(8) cells/mL, the cells reached the stationary phase with a concentration of about 5 x 10(8) cells/mL after 20 h, under both aerobic and anaerobic conditions. The distribution of the cell length did not vary significantly during the growth period, and both aerobically and anaerobically growing cells showed a similar cell length distribution. Although the cells showed similar growth patterns under both conditions, the anaerobically grown cells exhibited higher motility and magnetosensitivity. Actively growing cells under anaerobic conditions had an average swimming speed of 49 microm/s with a standard deviation of 20 microm/s. When the anaerobically growing cells were exposed to various concentrations of toxic compounds, such as 1-propanol and acetone, the swimming speed decreased with an increased concentration of the toxic compound. Accordingly, the relationship between swimming speed and toxicity can be used as an effective quantitative toxicity measurement; furthermore, the relative sensitivity of the proposed system was comparable to Microtox, which is commercially available. PMID- 11400103 TI - A metabolic model for acetate uptake under anaerobic conditions by glycogen accumulating organisms: Stoichiometry, kinetics, and the effect of pH. AB - A metabolic model for the stoichiometry of acetate uptake under anaerobic conditions by an enriched culture of glycogen accumulating organisms (GAOs) was developed and tested by experimental studies. Glycogen served as the source of both reducing power and energy to drive the process of acetate uptake. The amount of glycogen consumed and poly-beta-hydroxyvalerate (PHV) accumulated in the cells increased with increasing pH, indicating that the energy requirements for acetate uptake increased with pH. The composition of the accumulated poly-beta hydroxyalkanoates (PHAs) was adequately predicted using the assumption that acetyl-CoA and propionyl-CoA condense randomly to produce PHA. In addition, the rate of acetate uptake was strongly affected by the pH. The rate decreased with increasing pH and this dependence could be described with a saturation type of expression. A comparison of the rate of acetate uptake at low pH with the rates observed in enriched cultures of phosphorus accumulating organisms (PAOs) indicated that GAOs are able to compete effectively with PAOs in nutrient removal systems under certain conditions. PMID- 11400104 TI - Stoichiometry and kinetics of acetate uptake under anaerobic conditions by an enriched culture of phosphorus-accumulating organisms at different pHs. AB - The effect of pH on the stoichiometry and kinetics of acetate uptake by phosphorus-accumulating organisms (PAOs) was studied. The stoichiometry of glycogen consumption and poly-beta-hydroxy-alkanoates (PHA) accumulation was independent of the pH over the range 6.5 to 8.0. It was again demonstrated that the amount of phosphorus released per acetate taken up (P/Hac ratio) was linearly dependent on pH, because of additional energy requirements for acetate transport at higher pH. The slope of this relationship was similar to that in previously published work, but the absolute values were different, indicating that the P/Hac ratio is the most variable stoichiometric parameter associated with the anaerobic metabolism of PAOs. A kinetic expression for acetate-uptake rate was developed and tested. It assumes a zero-order form when the polyphosphate content of the biomass is not limiting. When the polyphosphate content becomes low, the rate is significantly decreased. The expression was tested in situations in which polyphosphate was a limiting factor in the rate of acetate uptake, in which the glycogen content of the biomass became very low, and in which both glycogen and polyphosphate were present in excess. The model was able to simulate the three situations adequately. Additionally, the rate of acetate uptake was independent of the pH for the range studied (6.5 to 8.0). PMID- 11400105 TI - Indirect measurement of water content in an aseptic solid substrate cultivation pilot-scale bioreactor. AB - A lack of models and sensors for describing and monitoring large-scale solid substrate cultivation (SSC) bioreactors has hampered industrial development and application of this type of process. This study presents an indirect dynamic measurement model for a 200-kg-capacity fixed-bed SSC bioreactor under periodic agitation. Growth of the filamentous fungus Gibberella fujikuroi on wheat bran was used as a case study. Real data were preprocessed using previously reported methodology. The model uses CO2 production rate and inlet air conditions to estimate average bed water content and average bed temperature. The model adequately reproduces the evolution of the average bed water content and can therefore be used as an on-line estimator in pilot-scale SSC bioreactors. To obtain a reasonable fit of the bed temperature, however, inlet air humidity measurements will have to be adjusted with a data reconciliation algorithm. Good estimation of temperature is important for the future design of improved water content estimation using state observers. The model also provides insight into understanding the complex behavior of the dynamic system, which could prove useful when establishing advanced model-based operational and control strategies. PMID- 11400106 TI - Relationship between cell proliferation, cell-cycle phase, and retroviral vector production in FLYRD18 human packaging cells. AB - The relatively low concentrations of retroviral vectors produced by most packaging cells requires the optimization and intensification of their production to make a commercially viable product for gene therapy. While a number of reports exist concerning target cell-cycle effects on retroviral vector infection efficiency, no studies have been reported on the effects of packaging cell cycle on vector production. We have studied the effect of proliferation of the human packaging cell line, FLYRD18, on vector production. In addition, the titer levels of vector produced by cells in each phase of the cell cycle were compared. Numerous studies suggested progression of the cells through the cell cycle to be essential for vector production. However, vector release was found not to be predominant in any particular phase of the cell cycle. These findings indicate that packaging cell proliferation is important for optimal virus production and that arrest of the cells in any particular phase of the cell cycle affords no benefits in retroviral vector production. In contrast to previous reports (using other cell lines), we observed no temporary inhibition of cell cycle progression after detachment of cells from their substratum and that virus production occurred immediately after re-plating of the cells. The findings in this report are important for determining the optimal culture conditions for vector production by packaging cells in vitro. PMID- 11400107 TI - Toluene biofiltration by the fungus Scedosporium apiospermum TB1. AB - The performance of biofilters inoculated with the fungus Scedosporium apiospermum was evaluated. This fungus was isolated from a biofilter which operated with toluene for more than 6 months. The experiments were performed in a 2.9 L reactor packed with vermiculite or with vermiculite-granular activated carbon as packing material. The initial moisture content of the support and the inlet concentration of toluene were 70% and 6 g/m3, respectively. As the pressure drop increased from 5-40 mm H2O a strong initial growth was observed. Stable operation was maintained for 20 days with a moisture content of 55% and a biomass of 33 mg biomass/g dry support. These conditions were achieved with intermittent addition of culture medium, which permitted a stable elimination capacity (EC) of 100 g/m3(reactor)h without clogging. Pressure drop across the bed and CO2 production were related to toluene elimination. Measurement of toluene, at different levels of the biofilter, showed that the system attained higher local EC (200 g/m3(r)h) at the reactor outlet. These conditions were related to local humidity conditions. When the mineral medium was added periodically before the EC decreases, EC of approximately 258 g/m3(r)h were maintained with removal efficiencies of 98%. Under these conditions the average moisture content was 60% and 41 mg biomass/g dry support was produced. No sporulation was observed. Evaluation of bacterial content and activities showed that the toluene elimination was only due to S. apiospermum catabolism. PMID- 11400108 TI - Optimization of maltodextrin hydrolysis by glucoamylase in a batch reactor. AB - The hydrolysis of maltodextrins (10 DE) by glucoamylase was studied in a batch reactor at temperatures between 40 and 80 degrees C and substrate concentration range from 17 to 300 kg/m(-3). The experimental data were fitted to a model including thermal deactivation of the enzyme. In the model, the reaction rate was correlated with an extended Michaelis-Menten equation including inhibition by product, and the thermal deactivation of glucoamylase was fitted with a first order reaction. The dependence of rate parameters on temperature was correlated using the Arrhenius equation. The differential equation of the model was integrated and the optimal enzyme demand and temperature were determined for isothermal operation. PMID- 11400109 TI - Possible complication regarding phosphorus removal with a continuous flow biofilm system: diffusion limitation. AB - Diffusion limitation of phosphate possibly constitutes a serious problem regarding the use of a biofilm reactor for enhanced biological phosphorus removal. A lab-scale reactor for simultaneous removal of phosphorus and nitrate was operated in a continuous alternating mode of operation. For a steady-state operation with excess amounts of carbon source (acetate) during the anaerobic phase, the same amount of phosphate was released during the anaerobic phase as was taken up during the anoxic phase. The measured phosphorus content of the biomass that detached during backwash after an anoxic phase was low, 2.4 +/- 0.4% (equal to 24 +/- 4 mg P/g TS). A simplified computer model indicated the reason to be phosphate diffusion limitation and the model revealed a delicate balance between the obtainable phosphorus contents of the biomass and operating parameters, such as backwash interval, biofilm thickness after backwash, and phase lengths. The aspect of diffusion is considered of crucial importance when evaluating the performance of a biofilter for phosphate removal. PMID- 11400110 TI - Glutamate synthesis via photoreduction of NADP+ by photostable chlorophyllide coupled with polyethylene-glycol. AB - Chlorophyllide a was coupled with alpha-(3-aminopropyl)-omega methoxypoly(oxyethylene) (PEG-NH2) to form a PEG-chlorophyllide conjugate through an acid-amide bond. The conjugate catalyzed the reduction of methylviologen in the presence of 2-mercaptoethanol. It also catalyzed the photoreduction of NADP+ or NAD+ in the presence of ascorbate as an electron donor and ferredoxin-NADP+ reductase as the coupling enzyme. Utilizing the reducing power of NADPH generated by PEG-chlorophyllide conjugate under illumination, glutamate was synthesized from 2-oxoglutarate and NH4+ in the presence of glutamate dehydrogenase. PEG chlorophyllide conjugate was quite stable toward light illumination compared with chlorophyll a. The increase in the molecular weight of PEG in the PEG chlorophyllide conjugates was accompanied by the enhancement of photostability of the conjugate and also by the increased solubility in the aqueous solution. PMID- 11400111 TI - Analysis of the potential contribution of estrogen receptor (ER) beta in ER cytosolic assay of breast cancer. AB - Estrogen receptor (ER) content is the most useful parameter for predicting hormone response therapy in breast cancer. Assays available for detecting ER in breast tumor cytosol are ligand-binding assay (LBA), which detects both ERalpha and ERbeta, and the enzymatic immunoassay (EIA), in which monoclonal antibodies are directed against ERalpha. As shown in several studies, the 2 assays correlate and both are used routinely. However, some discrepancies between the 2 assays were found and explanations remain controversial. We evaluated ERalpha and ERbeta mRNA coexpression in breast tumors in order to study whether the presence of ERbeta could account for differences between LBA and EIA in the determination of ER protein level. Using HeLa cell lines transfected with either ERalpha or ERbeta, we confirmed that EIA, using H222 and D547 monoclonal antibodies, recognizes only ERalpha expression, whereas LBA detects both isoforms. In 119 breast tumor cytosols, the correlation between ER-EIA and ER-LBA was high (r = 0.72), although some discrepancies were found. When analyzing ER mRNA expression of samples with higher LBA values, no overexpression of ERbeta mRNA relatively to ERalpha mRNA were observed. There was a difference in ERbeta/ERalpha ratio between ER-negative and ER-positive samples, with a 10-fold increased median ratio in ER-negative samples (p = 0.01). We thus confirmed that the major form of ER in breast cancer is the ERalpha at both the protein and mRNA levels. Moreover, our data do not support the hypothesis that ERbeta expression could explain differences between LBA and EIA in the determination of ER protein level. PMID- 11400112 TI - Cyclin D1-CDK4 complex, a possible critical factor for cell proliferation and prognosis in laryngeal squamous cell carcinomas. AB - Cyclin D1 and its catalytic partner CDK4 are known to play important roles in the G1/S checkpoint of the cell cycle. The complex formed by CDK4 and cyclin D1 has been strongly implicated in the control of cell proliferation and prognoses in human malignancies. We investigated the immunohistochemical expression of cyclin D1, CDK4 and proliferating cell nuclear antigen (PCNA) in 102 patients with laryngeal squamous cell carcinoma (LSCC). Cyclin D1 overexpression was observed in 59 cases (57.8%) of LSCC, and was significantly correlated with tumor site, tumor size, lymph node metastasis and advanced stage. CDK4 overexpression was observed in 48 cases (47.1%), and was significantly correlated with tumor size and advanced stage. Cyclin D1 and CDK4 expression was significantly associated with cell proliferation measured by PCNA (r = 0.812, p < 0.0001 and r = 0.725, p < 0.0001, respectively). The Kaplan-Meier analysis showed that cyclin D1 overexpression was significantly associated with disease-free survival and overall survival. CDK4 overexpression was significantly associated with overall survival. When cyclin D1 and CDK4 are combined, the patients with co overexpression of cyclin D1-CDK4 revealed the poorest overall survival. Additionally, in early-stage (I-II) cases, co-overexpression of cyclin D1-CDK4 was also revealed to possess a significant prognostic role. By multivariate analysis, cyclin D1 overexpression, lymph node metastasis and advanced stage were independent prognostic factors for disease-free survival. Cyclin D1 overexpression, CDK4 overexpression, tumor grade, lymph node metastasis and advanced stage were independent prognostic factors for overall survival. These findings indicate that cyclin D1 and CDK4 overexpression and/or co-overexpression of these proteins may play a pivotal role in the biological behavior of LSCC and may provide a strong prognostic implication. PMID- 11400113 TI - Expression of angiomodulin (tumor-derived adhesion factor/mac25) in invading tumor cells correlates with poor prognosis in human colorectal cancer. AB - Angiomodulin (tumor-derived adhesion factor/mac25/insulin-like growth factor binding protein-7), a cell-adhesive glycoprotein, is secreted by cancer cells and vascular endothelial cells. It may be involved in angiogenesis and modulation of the vascular functions necessary for tumor development. Although angiomodulin is expressed in colon cancer, there is limited information on it concerning cancer progression. In the present immunohistochemical study, we examined expression of angiomodulin in human colorectal cancer and its relationship with prognosis. A group of 89 surgically resected colorectal cancers was investigated immunohistochemically. In 37 cases (41.6%), angiomodulin was expressed in invading cancer cells. Early recurrence within 12 months after surgery was higher in patients with angiomodulin-expressing cancer than in those without (p < 0.05). The Kaplan-Meier life table revealed that patients with angiomodulin-positive tumor cells had a shorter survival time than those with negative cells (p < 0.01). The prognosis of patients with Dukes' C and angiomodulin-positive cells was apparently worse than that of patients with Dukes' D and angiomodulin negative cells. Multivariate analysis with logistic regression indicated that only angiomodulin expression in cancer cells, lymph node metastasis and age remained significant prognostic variables for survival (p < 0.05). Angiomodulin showed correlations with poor prognosis, indicating that it may be a useful prognostic marker in patients with colorectal cancer. PMID- 11400114 TI - Molecular analyses of the mitotic checkpoint components hsMAD2, hBUB1 and hBUB3 in human cancer. AB - During the metaphase-anaphase transition, the spindle checkpoint prevents segregation of chromosomes if the spindle assembly is perturbed. Critical components of this checkpoint are the MAD and BUB families of proteins, which prevent the proteolysis of Pds1 and B cyclins, producing mitotic arrest. In the present study, we first intended to resolve the role of the hsMAD2 gene in human cancer by determining the potential presence of hsMAD2 mutations in 44 primary bladder tumors, 42 soft-tissue sarcomas and 10 hepatocellular carcinomas. The entire coding region of the hsMAD2 gene was analyzed using PCR-SSCP and sequencing. One of the bladder tumor samples showed a point mutation consisting of a transition, ATC-->GTC (Ile-->Val) in codon 190 of hsMAD2. However, no differences were found in the mitotic arrest between cells transfected with mutant and wild-type MAD2 cDNA. We also identified mobility shifts in hsMAD2 in both normal and tumor DNA in 3 bladder tumors, 3 soft-tissue sarcomas and 1 hepatocellular carcinoma, consistent with a polymorphism at codon 143, CCA-->CCG (Pro-->Pro). Another polymorphism was identified in a hepatocellular carcinoma case at codon 22, GAG-->GAA (Glu-->Glu). In addition, a subgroup of 67 primary tumors was analyzed by Southern blot hybridization. No deletion or visible re arrangements were detected by comparing tumor and normal DNA band signals. Two other important components of the spindle mitotic checkpoint, hBUB1 and hBUB3, were also screened for mutations: hBUB1 in 43 bladder tumors and 9 bladder cell lines and hBUB3 only in the cell lines. Two polymorphisms were found in hBUB1 at positions 144, CAG-->CAA (Gln-->Gln) in 1 primary tumor and 1 bladder cell line, and 913 (ATC-->ATT, Ile-->Ile) in 1 primary tumor. We did not find sequence alterations in hBUB3. These results suggest that mutations of the hsMAD2, hBUB1 and hBUB3 genes are very rare in bladder tumors and that hsMAD2 alterations are also infrequent in soft-tissue sarcomas and hepatocellular carcinomas. PMID- 11400115 TI - Evaluation of chemotherapy response using technetium-99M-sestamibi scintigraphy in untreated adult malignant lymphomas and comparison with other prognosis factors: a preliminary report. AB - The purpose of this study was to predict chemotherapy response using technetium 99m methoxyisobutylisonitrile (Tc-MIBI) scintigraphy in untreated adult malignant lymphomas (ML) and compare the response with other prognosis factors. Before chemotherapy, 25 adult patients with ML were enrolled in this study. Tc-MIBI scintigraphy was performed 10 min after intravenous injection of Tc-MIBI to calculate tumor-to-background (T/B) uptake ratio and interpret the results visually. Chemotherapy response was evaluated in the first 1 to 2 years after completion of treatment by clinical and radiological methods. The mean T/B uptake ratio of the 15 patients with good response (3.3 +/- 0.6) was significantly higher than that of the 10 patients with poor response (1.2 +/- 0.1). All of the 15 patients with good response had positive Tc-MIBI scintigraphic results. All of the 10 patients with poor response had negative Tc-MIBI scintigraphic results. However, there were no significant differences in the incidences of good and poor responses for other prognosis factors. In our preliminary study, when compared with other prognosis factors, Tc-MIBI scintigraphy was the best tool to predict chemotherapy response in adult patients with ML. PMID- 11400116 TI - Heterogeneous distribution of P53 immunoreactivity in human lung adenocarcinoma correlates with MDM2 protein expression, rather than with P53 gene mutation. AB - Although the tumor suppressor p53 protein (P53) immunoreactivity and its gene (p53) mutation were reported to be significant prognostic indicators for human lung adenocarcinomas, little is known regarding the relationship between the heterogeneous distribution of P53 and its genetic status in each tumor focus and the clinicopathological significance. To determine how P53 is heterogeneously stabilized in patients, we compared P53 expression to both the p53 allelic mutation in exon 2 approximately 9 by polymerase chain reaction-single strand conformation polymorphism using microdissected DNA fractions, and the immunohistochemical MDM2 expression. Of the 48 positive to P53 in 118 lung adenocarcinomas examined, 10 with heterogeneous P53 expression were closely examined. The higher P53 expression foci in 7 of 10 cases were less differentiated, histologically in respective cases, and were frequently associated with fibrous stroma. Two had genetic mutations in exon 7 of the p53 gene in both the high and low P53 expression foci of cancer tissue indicating no apparent correlation between heterogeneous P53 expression and the occurrence of gene mutation. Immunohistochemical expression of MDM2 was significantly lower in high P53 expression areas (p < 0.05, the mean labeling indices of high and low P53 expression areas being 4.2 +/- 5.4% and 13.6 +/- 12.2%, respectively). In addition, among all the 118 cases examined, MDM2 expression was significantly suppressed in cases of p53 gene mutation, simultaneously with P53 overexpression, as compared with cases without both the p53 mutation and expression (p < 0.001). These findings suggest that the heterogeneous stabilization of P53 in human lung adenocarcinomas could be partly due to suppressed MDM2 expression. The overexpression of non-mutated P53 may afford a protective mechanism in human lung adenocarcinomas. PMID- 11400117 TI - Genetic polymorphisms of XRCC1 and risk of the esophageal cancer. AB - A variety of environmental factors were identified to be associated with the risk of esophageal cancer. The variation in capacity of DNA repair might influence environmental chemical-associated carcinogenesis. We hypothesized that the polymorphic XRCC1 genes might modify cancer susceptibility of the esophagus. To investigate the effect of XRCC1 genetic polymorphisms on codons 194, 280 and 399, we evaluated data from 105 patients of esophageal squamous cell carcinoma and 264 healthy controls, matching with age (+/-3 years), gender and ethnicity. The distribution of the 3 genotypes were not significantly different among patients and controls. However, among alcohol drinkers, the XRCC1399 Arg/Arg genotype was more frequently found in patients with esophageal cancer. After adjustment with other environmental confounders, the OR for the genotype of XRCC1399 Arg/Arg was 2.78 (95% CI =1.15-6.67) as compared with the XRCC1(399) Arg/Gln and XRCC1(399) Gln/Gln genotypes in the alcohol drinkers. Similar trends were observed among cigarette smokers and areca chewers. However, they did not reach a statistical significance. Our findings suggest that the polymorphic XRCC1 genes might modify the risk of alcohol-associated esophageal cancers. PMID- 11400118 TI - Phosphorylation of ERK1/2 mitogen-activated protein kinase is associated with poor response to anti-hormonal therapy and decreased patient survival in clinical breast cancer. AB - It is believed that growth factor phosphorylation cascades interact closely with oestrogen receptor (ER) signaling to regulate breast cancer growth, and that alterations in these pathways may underlie resistance to anti-hormones such as tamoxifen. There is an increasing body of experimental evidence implicating the mitogen-activated protein kinase extracellular signal-regulated-kinases ERK1 and ERK2 (ERK1/2 MAPK) in these events. The present study is the first to address the relationship between ERK1/2 MAPK phosphorylation (p-MAPK) and response to anti hormonal agents in clinical breast cancer (n = 90). Immunocytochemical analysis using a phosphorylation state-specific ERK1/2 MAPK antibody revealed 72% of breast tumors to have considerable nuclear p-MAPK immunostaining (designated p MAPK positive), whereas staining was barely detectable or absent in the remaining 28% (designated p-MAPK negative). Comparison with staining in normal breast material obtained from reduction mammoplasty patients (n = 10) demonstrated an increased frequency of higher intensity p-MAPK immunostaining cells within carcinomas (p = 0.002). Significant relationships were revealed between p-MAPK positivity and poorer quality (p = 0.001) and shortened duration (p = 0.006) of anti-hormonal response, as well as with decreased survival time from the initiation of therapy (p = 0.022). These associations were retained in ER positive disease (p = 0.013, p = 0.037 and p = 0.048 respectively), where multivariate analysis demonstrated p-MAPK status to be a significantly independent predictor for response duration (p = 0.034) and patient survival (p = 0.029). Phosphorylated ERK1/2 MAP kinase is thus potentially prognostic for prediction of response to anti-hormonal agents and survival, data providing further evidence that ERK1/2 MAP kinase plays a role in circumvention of anti hormonal response in breast cancer. PMID- 11400119 TI - Decreased expression of BRCA2 mRNA predicts favorable response to docetaxel in breast cancer. AB - The clinical usefulness of BRCA1 and BRCA2 mRNA levels in tumor tissues in the prediction of response to docetaxel (DOC) treatment has been studied in breast cancer patients. Twenty-five patients with locally advanced breast tumors (n = 13) or locally recurrent tumors (n = 12) underwent tumor biopsy and were treated with DOC (60 mg/m2 every 3 weeks). BRCA1 and BRCA2 mRNA levels in the tumors were determined by real-time PCR, and the expression of 6 biological markers (P glycoprotein, p53, erbB2, BCL2, MIB1, estrogen receptor-alpha) in the tumors was determined by immunohistochemistry. BRCA2 mRNA levels (0.547 +/- 0.200, mean +/- SE) of responders to DOC treatment were significantly (p < 0.05) lower than those of non-responders (1.538 +/- 0.358), but there was no significant difference in BRCA1 mRNA levels between responders (0.389 +/- 0.081) and non-responders (0.779 +/- 0.172). Tumors were dichotomized into groups with high or low BRCA2 mRNA levels according to the cut-off value of 0.13. The response rate (25%) of tumors with high BRCA2 mRNA levels was significantly (p < 0.01) lower than that (100%) of tumors with low BRCA2 mRNA levels. Positive predictive value, negative predictive value and diagnostic accuracy of the BRCA2 mRNA assay in the prediction of response to DOC were 100%, 75% and 80%, respectively. No significant difference was found between responders and non-responders in the expression status of any of the other 6 biological markers. These results suggest that BRCA2 mRNA levels in tumor tissues might be clinically useful in the prediction of response to DOC treatment in breast-cancer patients. PMID- 11400120 TI - Intracystic epidermal growth factor level is predictive of breast-cancer risk in women with gross cystic disease of the breast. AB - Women affected by gross cystic disease of the breast have an increased risk of breast cancer. We report here the incidence of breast cancer by cyst type and intracystic epidermal growth factor (EGF) concentration. Our retrospective study included 504 women who had at least 1 cyst aspiration between 1985 and 1993. Cyst fluids were processed for electrolyte concentration (n = 378), EGF concentration (n = 347) or both (n = 337). Age-standardized incidence ratios (SIRs) were estimated using the population of the Genoa Cancer Registry. A multivariate Poisson regression model was used to estimate relative risks (RRs) when the study groups were compared directly. By June 1999, 19 invasive breast cancers had developed in the cohort of women. The age SIR of breast cancer calculated for the whole cohort was 3.32 (95% confidence interval 2.00-5.18). The ratio was not affected by age and was only moderately increased in women with a positive family history of breast cancer and type I cysts (i.e., those with a Na+/K+ ratio <3). However, it was significantly increased in women with high EGF concentrations. Direct comparisons confirmed that age, cyst type and family history only moderately increased the RR, whereas EGF concentration was a strong predictor of risk. Our results confirm that women affected by palpable cysts have an increased risk of developing breast cancer and suggest that the risk is higher in women with high intracystic EGF concentrations. PMID- 11400121 TI - c-erbB-2 (HER-2/neu) gene amplification is a better indicator of poor prognosis than protein over-expression in operable breast-cancer patients. AB - Our aim was to compare the prognostic value of c-erbB-2 gene amplification analyzed by Southern blot with that of protein (p185) over-expression measured by immunohistochemistry in 172 patients with operable breast cancer (BC). Amplification and p185 over-expression were found in 31 (18%) and 51 (30%) BCs, respectively. All but 1 of the tumors showed both amplification and over expression, while 21 (12%) tumors displayed over-expression without amplification. The risk of death associated with c-erbB-2 gene amplification and p185 over-expression was evaluated by multivariate analysis, taking into account tumor size, histoprognostic grade, hormone receptors and axillary node status. During a mean follow-up of 9.5 (+/-2) years, node involvement (p < 0.001), c-erbB 2 gene amplification (p = 0.02) and negative hormone receptors (p = 0.02) were found to be independent prognostic indicators of the risk of death. Over expression of p185 with no amplification was not correlated with this risk. When the risk of death associated with c-erbB-2 amplification was studied according to chemo- and hormone therapy, no significant difference was observed between subgroups of subjects. Amplification was also associated (p = 0.02) with the risk of multifocal distant metastases (i.e., metastases detected concomitantly in at least 2 sites) and, thus, with BC aggressiveness. These data show the importance of c-erbB-2 gene amplification in predicting the long-term outcome of patients and in selecting eligible patients for c-erbB-2-targeted therapies. PMID- 11400122 TI - Association of estrogen receptor and glucocorticoid receptor gene polymorphisms with sporadic breast cancer. AB - We have utilized a cross-sectional association approach to investigate sporadic breast cancer. Polymorphisms in 2 candidate genes, ESRalpha and GRL, were examined in an unrelated breast cancer-affected and age-matched control population. Several polymorphic regions within the ESRalpha gene have been identified, and some alleles of these polymorphisms have been found to occur at increased levels in breast-cancer patients. Additionally, variations in GRL have the potential to disrupt cell transcription and may be associated with cancer formation. We analyzed 3 polymorphisms, from codons 10 (TCT to TCC), 325 (CCC to CCG) and 594 (ACA to ACG) of ESRalpha, and a highly polymorphic dinucleotide repeat, D5S207, located within 200 kb of the GRL. When allelic frequencies of the codon 594 (exon 8) ESR polymorphism were compared between affected and unaffected populations, a significant difference was observed (p = 0.005). Results from the D5S207 dinucleotide repeat located near GRL also indicated a significant difference between the tested case and control populations (p = 0.001). Allelic frequencies of the codon 10 and codon 325 ESR polymorphisms were not significantly different between populations (p = 0.152 and 0.181, respectively). Our results indicate that specific alleles of the ESR gene (alpha subtype) and a marker for the GRL gene locus are associated with sporadic breast-cancer development in the tested Caucasian population and justify further investigation of the role of these and other nuclear steroid receptors in the etiology of breast cancer. PMID- 11400123 TI - The architecture of a collagen coating on a synthetic polymer influences epithelial adhesion. AB - The current study sought to identify a collagen coating methodology for application to polymer surfaces that would provide for the development of adhesive structures responsible for the sustained adhesion of corneal epithelial tissue. We compared an uncoated microporous polycarbonate surface and equivalent surfaces coated with either covalently immobilized collagen I or chemically crosslinked collagen I gel in a corneal explant outgrowth assay over 21 days. Electron microscopy was used to examine the formation of hemidesmosomes, basal lamina, and anchoring fibrils at the tissue-polymer interface. The crosslinked collagen gel preparation supported the overlying epithelial tissue across the pore openings and allowed for the formation of identifiable basal lamina, hemidesmosomes, and anchoring fibrils between the epithelial tissue and the polymer surface. Hemidesmosomal plaque, but no basal lamina or anchoring fibril formation, occurred on the uncoated surface or on that coated with covalently immobilized collagen I. We propose that the collagen matrix provided by the crosslinked collagen gel was reorganized by the epithelial tissue and that this, combined with the secretion of ECM molecules, served to limit the diffusion of basement membrane components, which permitted an increase in the local concentration of these molecules, which favored the assembly of epithelial adhesive structures. PMID- 11400124 TI - Microtubular architecture of biodegradable polymer scaffolds. AB - It is a relatively new approach to generate tissues with mammalian cells and scaffolds (temporary synthetic extracellular matrices). Many tissues, such as nerve, muscle, tendon, ligament, blood vessel, bone, and teeth, have tubular or fibrous bundle architectures and anisotropic properties. In this work, we have designed and fabricated highly porous scaffolds from biodegradable polymers with a novel phase-separation technique to generate controllable parallel array of microtubular architecture. Porosity as high as 97% has been achieved. The porosity, diameter of the microtubules, the tubular morphology, and their orientation are controlled by the polymer concentration, solvent system, and temperature gradient. The mechanical properties of these scaffolds are anisotropic. Osteoprogenitor cells are seeded in these three-dimensional scaffolds and cultured in vitro. The cell distribution and the neo-tissue organization are guided by the microtubular architecture. The fabrication technique can be applied to a variety of polymers, therefore the degradation rate and cell--matrix interactions can be controlled by the chemical composition of the polymers and the incorporation of bioactive moieties. These microtubular scaffolds may be used to engineer a variety of tissues with anisotropic architecture and properties. PMID- 11400125 TI - Periodate-mediated glycosaminoglycan stabilization in bioprosthetic heart valves. AB - Bioprosthetic heart valves (BPHVs) derived from glutaraldehyde-crosslinked porcine aortic valves are frequently used in heart valve replacement surgeries. However, the majority of bioprostheses fail clinically because of calcification and degeneration. We have recently shown that glycosaminoglycan (GAG) loss may be in part responsible for degeneration of glutaraldehyde-crosslinked bioprostheses. In the present studies, we used a mild reaction of periodate-mediated crosslinking to stabilize glycosaminoglycans in the bioprosthetic tissue. We demonstrate the feasibility of periodate reaction by crosslinking major components of extracellular matrix of bioprosthetic heart valve tissue, namely type I collagen and hyaluronic acid (HA). Uronic acid assay of periodate-fixed HA collagen matrices showed 48% of HA disaccharides were bound to collagen. Furthermore, we show that such reactions are also feasible to fix glycosaminoglycans present in the middle spongiosa layer of bioprosthetic heart valves. The periodate reactions were compatible with conventional glutaraldehyde crosslinking and showed adequate stabilization of extracellular matrix as demonstrated by thermal denaturation temperature and collagenase assays. Moreover, uronic acid assays of periodate-fixed BPHV cusps showed 36% reduction in the amount of unbound GAG disaccharides as compared with glutaraldehyde crosslinked cusps. We also demonstrate that calcification of BPHV cusps was significantly reduced in the periodate-fixed group as compared with the glutaraldehyde-fixed group in 21-day rat subdermal calcification studies (periodate-fixed tissue Ca 72.01 +/- 5.97 microg/mg, glutaraldehyde-fixed tissue Ca 107.25 +/- 6.56 microg/mg). We conclude that periodate-mediated GAG fixation could reduce structural degeneration of BPHVs and may therefore increase the useful lifetime of these devices. PMID- 11400126 TI - Chemotactic penetration of keratocytes in ePTFE polymer in vitro. AB - Expanded polytetrafluoroethylene (ePTFE) is used as a support for artificial corneas. Implanted in corneas, most of the time this polymer is colonized by corneal host cells. The absence of colonization often coincides with extrusion of the polymer. Therefore, we decided to introduce keratocytes into ePTFE in vitro before implantation. Because keratocytes do not spontaneously enter ePTFE, we used several chemoattractants, separately and in a mixture, to stimulate the penetration of cultured keratocytes into the polymer. The influence of the passage number on cell penetration was also studied. No significant differences were observed up to the seventh passage, although seventh-passage cells penetrated somewhat more slowly than younger cells. Satisfactory results were obtained with four of the tested chemotactic factors: IL-6, type alpha transforming growth factor (TGF-alpha), platelet derived growth factor isoform BB (PDGF-BB), and fibroblast growth factor-2 (FGF-2). Under our experimental conditions, two to more than six million keratocytes entered the polymer discs with a volume of 706.5 mm(3) in the presence of these four chemoattractants. TGF alpha was the most efficient and was selected for further in vitro and in vivo studies. PMID- 11400127 TI - Interaction between topography and coating in the formation of bone nodules in culture for hydroxyapatite- and titanium-coated micromachined surfaces. AB - Rat osteoblast cultures were maintained from 24 h to 6 weeks on hydroxyapatite (HA)- or titanium (Ti)-coated smooth and micromachined grooved substrata in medium supplemented with L-ascorbic acid-2-phosphate and beta-glycerophosphate to promote mineralization. The HA coatings, approximately 1 microm thick, were characterized using X-ray diffraction, surface roughness, and scanning electron microscopy (SEM). Osteoblasts elongated, aligned, and moved in the direction of the grooves on both Ti and HA grooved surfaces. HA surfaces produced significantly more bone-like nodules than Ti surfaces. All grooved substrata produced significantly more nodules than smooth surfaces. These results are consistent with the hypothesis that substrata can increase osteogenesis by formation of an appropriate microenvironment. There was also a statistically significant interaction between topography and chemistry in the formation of mineralized nodules. A strong correlation (r = 0.958) between alkaline phosphatase (Alk-P) at 2 weeks and nodule counts at 6 weeks was observed, suggesting that Alk-P may possibly be used as a leading indicator of osteogenesis on microfabricated surfaces. The results of this study indicate that surface topography and chemistry can affect osteogenesis, and that interactions between chemistry and topography can occur. PMID- 11400128 TI - Fabrication of porous calcium polyphosphate implants by solid freeform fabrication: a study of processing parameters and in vitro degradation characteristics. AB - Solid freeform fabrication (SFF) involves the creation of a solid 3-D object of desired shape by successively adding raw materials in particles or layers. Its use in fabricating surgical implants is being explored. The objective of this study was to determine the feasibility of using SFF to build porous parts of calcium polyphosphate (CPP), a linear condensed phosphate that has been suggested as a material for forming bioresorbable skeletal replacement implants. CPP powders (<25 microm in particle size) were added to an UV curable monomer (SOMOS 6110) at a solids loading of 25 vol %, with the addition of a commercial dispersant to prevent particle agglomeration and settling. Viscosity and cure depth measurements were performed to insure that CPP suspension met the requirements deemed necessary for use in SFF. The CPP suspension was bulk cured and sintered in molds in order to assess binder removal and sintering parameters. Using a three-point bend test, the ultimate bending strength and energy-to fracture of sintered CPP samples simulating parts to be formed by this strategy were characterized. In vitro degradation studies using 0.1M of tris-buffered solution were performed to assess the effect of aging on mechanical properties of the samples as a function of the processing route and resulting structures. The polymer binder successfully was removed from the cured ceramic suspension by developing a procedure that combined slow heating rates with low temperature dwells. Sintering CPP at 585 degrees C for 1 h produced amorphous material samples with average porosity of 27.7 +/- 2.0%. Sintering CPP at 600 degrees C for 1 h produced a crystalline material with samples having an average porosity of 22.9 +/- 1.3%. Crystalline CPP was found to exhibit superior bend strength and toughness compared with amorphous CPP. Both samples experienced a decline in mechanical properties during in vitro degradation; however, the effects were more pronounced with the amorphous CPP samples. Amorphous CPP was found to degrade four times faster than crystalline CPP, as shown by high levels of phosphate present in the degradation solution and a noticeable increase in the porosity of the samples. Crystalline CPP was more resistant to attack as dissolution was limited to surface features of the sintered particles. PMID- 11400129 TI - Enzyme-induced biodegradation of polycarbonate polyurethanes: dependence on hard segment concentration. AB - Polycarbonate-based polyurethanes with varying hard segment contents were synthesized. The physical and chemical structures were characterized by using gel permeation chromatography, differential scanning calorimetry, water uptake testing, Fourier transform infrared, and attenuated total reflectance--Fourier transform infrared. The polymers were incubated with cholesterol esterase in a phosphate buffer solution at 37 degrees C over 10 weeks. A higher resistance to hydrolytic degradation was observed in polycarbonate-based urethanes with higher hard segment content. The analysis of the material structures revealed that the degradation of polycarbonate-based urethanes was preferentially initiated at non hydrogen-bonded carbonates and urethanes. Although the crystallinity of the polycarbonate soft segment may contribute to reducing the hydrolytic degradation catalyzed by cholesterol esterase, it was found to be relatively minor in comparison to the importance of hydrogen bonding between the carbonate and urethane groups. These observations suggest that the biostability of polyurethanes and specifically polycarbonate-based polyurethanes can be improved by manipulating the degree of hydrogen bonding within the materials. PMID- 11400130 TI - Modification of fibrinogen with poly(ethylene glycol) and its effects on fibrin clot characteristics. AB - The suitability of existing topical fibrin glue preparations for tissue sealing or local drug delivery applications is greatly limited by their poor mechanical properties and the limited capacity of fibrinogen (Fgn) to actively bind growth factors or other therapeutic agents. Poly(ethylene glycol) (PEG) offers potential solutions to these problems by providing a mechanism for increasing the number of crosslinks between adjacent fibrin monomer molecules or for covalently crosslinking Fgn to therapeutic agents. The feasibility of this approach requires the full biological activity, or clottability, of PE glycolated Fgn. This study characterizes the clot characteristics of Fgn modified to varying degrees with monofunctional succinimidyl propionate PEG (5000 Da). The data indicate that, although thrombin clotting times are significantly altered, Fgn maintains 90% of its capacity to clot upon the addition of up to 5 PEG/Fgn. Further derivatization significantly decreases the Fgn clottability. The addition of up to 5 PEG/Fgn has little, if any, effect on the kinetics of degradation by plasmin. The results suggest that limited modification of Fgn with lysine-reactive PEG allows therapeutic enhancement of fibrin glues. PMID- 11400131 TI - Heparin-carrying polystyrene (HCPS)-bound collagen substratum to immobilize heparin-binding growth factors and to enhance cellular growth. AB - Heparin-carrying polystyrene (HCPS), consisting of low molecular weight heparin chains linked to a synthetic polystyrene core, is able to attach to polymeric surfaces. In this study, HCPS has efficiently bound to collagen-coated micro plates and collagen membranes thereby retaining the binding of heparin-binding growth factors, such as vascular endothelial growth factor (VEGF)(165) or fibroblast growth factor (FGF)-2. Both human skin fibroblast cells and human umbilical vein endothelial cells have shown a good adherence to both collagen- and HCPS-bound collagen substrata. The growth rate of the fibroblast cells on the HCPS-bound collagen substratum in the presence of low concentrations of FGF-2 is higher than on a collagen surface. The fibroblast cells grow at a significantly higher rate on the HCPS-bound collagen substratum retained with FGF-2. Similarly, the growth rate of the endothelial cells on the HCPS-bound collagen substrata in the presence of low concentrations of either FGF-2 or VEGF(165) is higher than on collagen. The endothelial cells also grow at a significantly higher rate on the HCPS-bound collagen substratum retained with either FGF-2 or VEGF(165). These results indicate that HCPS-bound collagen substrata with various bioactive heparin-binding molecules may provide novel biomaterials controlling cellular activities such as growth and differentiation. PMID- 11400132 TI - Vascular graft endothelialization: comparative analysis of canine and human endothelial cell migration on natural biomaterials. AB - Canines are typically used as the standard preclinical model to gauge the success of vascular graft materials. However, canines spontaneously re-endothelialize vascular grafts, whereas humans do not, even after years. This raises questions of why there are differences in vascular healing between humans and other species and whether the canine is the appropriate preclinical model. In the present study we evaluated human and canine endothelial cell (EC) migration on the novel cross linked collagen biomaterial PhotoFix(TM) pericardium. We compared in vitro migration of these cells on PhotoFix alone and on PhotoFix adsorbed with various growth factors (aFGF and bFGF) and adhesion proteins (fibronectin, collagen IV, vitronectin, and laminin). We also compared human and canine ECs in terms of their morphologies and prostacyclin production. We found that human umbilical vein ECs (HUVECs) and canine ECs (CECs) migrated well on PhotoFix, suggesting that this biomaterial may be a good vascular graft candidate. Both cell types responded similarly to different growth factors and adhesive proteins, but HUVEC migration was consistently higher than that for CECs. This suggested that human in vivo graft re-endothelialization is likely not hindered by poor endothelial migration but is hindered by other cellular or graft properties. PMID- 11400133 TI - Release of glycosaminoglycans in physiological saline and water by wet-spun chitin--acid glycosaminoglycan fibers. AB - The present study sought to prepare novel chitin- acid glycosaminoglycan fibers that released a portion of the glycosaminoglycans in animal body fluids for use as novel biocompatible dressing materials (artificial skin) in the veterinary and clinical fields. A clear solution of sodium N-acetylchitosan salt (alkaline chitin) mixed with sodium hyaluronate, sodium heparin, sodium chondroitin 4 sulfate, sodium chondroitin 6-sulfate, or sodium dermatan sulfate in 14% aqueous NaOH was spun through a viscose-type spinneret into a 10% aqueous H(2)SO(4) solution saturated with (NH(4))(2)SO(4) at room temperature. The result was chitin fibers containing 5--33% glycosaminoglycans. In a dry state these fibers were white and had a soft feel but they were mechanically weak (0.31--0.69 g/denier tenacity and 3.1--10.6% elongation). Portions (85--97%) of the glycosaminoglycans were released from the fibers by soaking them in physiological saline or distilled water. Scanning electron microscopy analyses revealed a smooth striped surface on the original filaments, and a scaly surface appeared on the chitin filament after soaking. PMID- 11400134 TI - Bioactive titanium: effect of sodium removal on the bone-bonding ability of bioactive titanium prepared by alkali and heat treatment. AB - As reported previously, bioactive titanium is prepared by simple alkali and heat treatment, and can bond to living bone directly. The purpose of this study was to accelerate the bioactivity of bioactive titanium in vivo. In in vitro study, sodium removal by hot water immersion enhanced the apatite-forming ability of bioactive titanium in simulated body fluid dramatically. The specific anatase structure of titania gel was effective for apatite formation in vitro. In the current study, we investigated the in vivo effect of sodium removal on the bone bonding strength of bioactive titanium. Sodium-free bioactive titanium plates were prepared by immersion in an aqueous solution of 5 M NaOH at 60 degrees C for 24 h, followed by immersion in distilled water at 40 degrees C for 48 h before heating them at 600 degrees C for 1 h. Three kinds of titanium plates were inserted into rabbit tibiae, including untreated cp-Ti, conventional alkali- and heat-treated Ti, and sodium-free alkali- and heat-treated Ti. In vivo bioactive performance was examined mechanically and histologically after 4, 8, 16, and 24 weeks. Sodium removal enhanced the bone-bonding strength of bioactive titanium at 4 and 8 weeks postoperatively; however, its bone-bonding strength was inferior to that of conventional alkali- and heat-treated titanium at 16 and 24 weeks. Histological examinations after the detaching test revealed breakage of the treated layer in the sodium-free alkali- and heat-treated titanium group. In conclusion, sodium removal accelerated the in vivo bioactivity of bioactive titanium and achieved faster bone-bonding because of its anatase surface structure, but the loss of the surface's graded structure due to the complete removal of sodium decreased the adhesive strength of the treated layer to the titanium substrate. Further investigations are required to determine the optimum conditions for preparation of bioactive titanium. PMID- 11400135 TI - Bioactive bone cement: effects of phosphoric ester monomer on mechanical properties and osteoconductivity. AB - A new bioactive bone cement, designated GBC, has been developed. It consists of polymethyl methacrylate (PMMA) as an organic matrix and bioactive glass beads as an inorganic filler. The bioactive beads, consisting of MgO--CaO--SiO(2)- P(2)O(5)--CaF(2) glass, have been newly designed, and a novel PMMA powder was selected. The purpose of the present study was to evaluate the effects on mechanical properties and osteoconductivity of adding a phosphoric ester (PE) monomer to the cement as an adhesion-promoting agent. Four kinds of cements were prepared: GBC, GBC with PE (designated GBC/PE), a cement consisting of the same PMMA used in GBC with apatite- and wollastonite-containing glass-ceramic (AW-GC) powder (designated AWC), and AWC with PE (designated AWC/PE). Each filler was added to the cement at 70 wt %. Adding PE to either GBC or AWC resulted in increases in the bending strength and decreases in the Young's modulus compared with the unmodified cements. Cements were packed into the intramedullar canals of rat tibiae to evaluate osteoconductivity as determined by an affinity index. Rats were sacrificed at 4 and 8 weeks after operation. The affinity index (length of bone in direct contact with the cement expressed as a percentage of the total length of the cement surface) was calculated for each cement. Adding PE to either GBC or AWC resulted in significant increases in the affinity index compared with the unmodified cements. The affinity index for GBC was significantly higher than that of AWC, and that for GBC/PE was also significantly higher than that of AWC/PE. The affinity indices for each cement increased significantly with time up to 8 weeks. Our study revealed that the higher osteoconductivity of GBC/PE was due to the large alkyl group in the PE monomer, to the hydrophilicity of the phosphoric acid in the PE monomer, and to the higher bioactivity of the bioactive glass beads at the cement surface. GBC/PE shows promise as an alternative bone cement with improved properties compared with conventional PMMA bone cement. PMID- 11400136 TI - The effect of NaOCl dentin treatment on nanoleakage formation. AB - The term "nanoleakage" has been introduced to explain a penetration pathway within hybrid layers of the dentin-composite junction in the absence of gap formation. This phenomenon is argued in the literature to be a risk factor for the quality of the dentin bonding. NaOCl is a well-known agent used to remove collagen layers that are exposed after acid etching. The purpose of this study was to determine the influence of an NaOCl treatment of dentin to the formation of nanoleakage. Class V cavities were prepared in 40 human molars with the cervical margins located in dentin. After etching with phosphoric acid, 20 samples were treated with 10% NaOCl for 60 s, and 20 samples were used as control. Composite restorations were placed using two different bonding systems. The specimen were exposed in silver nitrate solution and then in developer. After embedding, the teeth were sectioned, dried, coated with gold, and analyzed by SEM. For all the specimen in the control group, backscatter images indicated a silver penetration within the hybrid layer. There was no penetration of silver detectable in the teeth that were treated with NaOCl. We conclude that the removal of the collagen layer by using NaOCl avoids the formation of nanoleakages with the materials tested. PMID- 11400137 TI - Identification and quantification of irradiation in UHMWPE through trans-vinylene yield. AB - trans-Vinylene unsaturations generated in ultrahigh-molecular-weight polyethylene (UHMWPE) after radiation treatment scales linearly with the absorbed dose level, and can serve as an internal dosimeter for determining the radiation dose level for sterilizing or crosslinking UHMWPE. We measured the trans-vinylene concentration using infrared spectroscopy and generated calibration curves for ultrahigh-molecular-weight polyethylene (ram-extruded GUR 1050) after cold e beam, cold gamma, and warm e-beam irradiations. The trans-vinylene content increased linearly with the absorbed dose level in all cases except with cold gamma irradiation at dose levels higher than approximately 70 kGy. At that dose level the trans-vinylene content of the warm irradiated samples was higher than those of the cold irradiated samples. Following postirradiation melting, the trans-vinylene content of the gamma irradiated samples increased. Such trans vinylene quantification can be an internal dosimeter for crosslinked UHMWPE when radiation is used for improving its wear resistance or sterilizing the device made thereof. It can also be utilized to assess the spatial uniformity of the absorbed radiation dose level. PMID- 11400138 TI - Ultrasonic characterization of the curing process of hydroxyapatite-modified bone cement. AB - Ultrasonic parameters such as velocity of sound and broad-band ultrasonic attenuation (BUA) are sensitive to changes in the viscoelastic properties of a material. Bone cement undergoes changes is these properties as it cures. By monitoring the propagation of ultrasonic pulses through a sample of curing bone cement, the curing reaction of polymethylmethacrylate-based (PMMA) bone cement was investigated for hydroxyapatite (HA) concentrations of 0, 10, and 30% (by weight). As the material hardens, the velocity of sound increases by 70%. BUA shows a large peak at the midpoint of the velocity transition. These data are used to compare the cure time and cure duration for PMMA bone cement mixed with hydroxyapatite particles. Measurements of the final sound velocity and BUA were also performed to investigate the mechanical properties of the fully cured cement, and to compare to compression testing data. This is the first time the curing process of bone cement has been investigated as a function of hydroxyapatite concentration. Results indicate that the cure time is not significantly affected by the addition of HA particles, and that both velocity of sound and BUA are sensitive to the curing process. PMID- 11400139 TI - A comparative biocompatibility study of microspheres based on crosslinked dextran or poly(lactic-co-glycolic)acid after subcutaneous injection in rats. AB - Microspheres based on methacrylated dextran (dex-MA), dextran derivatized with lactate-hydroxyethyl methacrylate (dex-lactate-HEMA) or derivatized with HEMA (dex-HEMA) were prepared. The microspheres were injected subcutaneously in rats and the effect of the particle size and network characteristics [initial water content and degree of methacrylate substitution (DS)] on the tissue reaction was investigated for 6 weeks. As a control, poly(lactic-co-glycolic)acid (PLGA) microspheres with varying sizes (unsized, smaller than 10 microm, smaller and larger than 20 microm) were injected as well. A mild tissue reaction to the PLGA microspheres was observed, characterized by infiltration of macrophages (MOs) and some granulocytes. Six weeks postinjection, the PLGA microspheres were still present. However, their size was decreased indicating degradation and many spheres had been phagocytosed. The tissue reaction was hardly affected by size differences, except for particles smaller than 10 microm, which induced an extensive tissue reaction. The initial tissue reaction to nondegradable dex-MA microspheres was stronger than towards the PLGA microspheres, but at day 10 the tissue reactions were comparable for both groups. Six weeks postinjection, the dex-MA microspheres were completely phagocytosed, and no signs of degradation were observed. The size and initial water content of dex-MA microspheres hardly affected the tissue response, although less granulocytes were observed for microspheres with higher DS. Slowly degrading dextran microspheres composed of dex-(lactate(1)-)HEMA induced a tissue reaction comparable to the PLGA microspheres. However, degradation of the dex-(lactate(1,3)-)HEMA microspheres was associated with an increased number of MO's and giant cells, both phagocytosing the microspheres and their degradation products. Similar to PLGA, no adverse reactions were observed for the nondegradable dex-MA and degradable dextran microspheres. This study shows that both nondegradable and degradable dextran-based microspheres are well tolerated after subcutaneous injection in rats, which make them interesting candidates as controlled drug delivery systems. PMID- 11400140 TI - Dysplasia classification: pathology in disgrace? AB - Grading of dysplasia is demanded almost daily from most diagnostically active pathologists. It is also notoriously subjective and lacks intra- and inter observer reproducibility. This is partly due to the lack of validated morphological criteria, upon which pathologists have reached consensus. It is largely due to the biological nature of the evolution of dysplasia, not in discrete steps but as a continuum. Better morphological definition, but also fundamental research into the nature of the process, is necessary to resolve this issue. PMID- 11400141 TI - Expression of cytokine and chemokine genes in Epstein-Barr virus-associated nasopharyngeal carcinoma: comparison with Hodgkin's disease. AB - Nasopharyngeal carcinoma (NPC) and Hodgkin's disease (HD) are characterized by their association with Epstein-Barr virus (EBV) and the presence of an intense lymphoid stroma, consisting of T lymphocytes and other reactive cells. In both entities, the tumour cells express viral proteins known to provide target epitopes for cytotoxic T-cells (CTLs), yet in vivo, the tumour cells appear to escape CTL recognition. A comparative in situ hybridization study of cytokine and chemokine gene expression in NPC and HD has been undertaken, focusing on cytokines which are known to be inducible by EBV in vitro. Hodgkin and Reed Sternberg (HRS) cells expressed interleukin (IL)-6, IL-8, and IL-10, and the thymus and activation regulated chemokine (TARC) in 15/22, 0/22, 5/22, and 16/21 cases, respectively. In NPC, the epithelial tumour cells showed expression of IL 6 in 3/43 cases and of IL-8 in 2/40 cases. There was no detectable expression of IL-10 and TARC in these cases. These data confirm that HRS cells frequently express cytokine and chemokine genes and suggest that this may enable HRS cells to modulate the immune response in their microenvironment and to escape CTL detection. In contrast, NPC tumour cells show only rare expression of IL-6 and IL 8 and no detectable expression of IL-10 and TARC. Thus, the results suggest that the mechanisms employed by the EBV-positive tumour cells to escape immune recognition and destruction differ between HD and NPC. PMID- 11400142 TI - Inter-observer variation between general and specialist gastrointestinal pathologists when grading dysplasia in ulcerative colitis. AB - Histological dysplasia is the cornerstone of colorectal cancer surveillance in ulcerative colitis (UC). Recently, pathologists have received unfavourable media attention concerning other cancer screening programmes. The aim of this study was to determine whether colonic biopsy specimens should be examined by gastrointestinal pathologists as opposed to generalists, by examining inter observer variation between the two groups. Fifty-one coded slides showing varying degrees of dysplasia were mailed to seven gastrointestinal and six general histopathologists. Pathologists allocated each biopsy into one of four categories without the benefit of a clinical history or an opportunity to use the 'indefinite' category that is included in the Riddell classification. The responses were analysed using kappa statistics. The overall kappa statistic for gastrointestinal pathologists was 0.30 [95% confidence interval (CI)=0.26-0.34] and for general pathologists 0.28 (95% CI=0.23-0.32). Agreement was best for high grade dysplasia (kappa of 0.54 and 0.61 for GI and general pathologists, respectively). There was total concordance of the 13 pathologists in only four of the 51 slides (7.8%) (95% CI=0.4-15.2%). It is concluded from these results that gastrointestinal pathologists are no better than generalists when grading dysplasia in UC and that agreement is poor in both groups. There is therefore no evidence that there would be any benefit in having specialist histopathology centres concentrating specifically on the interpretation of all surveillance colonoscopy biopsies from around the UK. It must be made clear to the public that surveillance and screening programmes carry a significant rate of histological error and that perfection cannot be expected or achieved with present methods. PMID- 11400143 TI - Expression of the tumour necrosis factor receptor-associated factors 1 and 2 in Hodgkin's disease. AB - The tumour necrosis factor receptor-associated factors (TRAFs) 1 and 2 participate in the signal transduction of various members of the tumour necrosis factor receptor (TNFR) family, including TNFR1, TNFR2, CD40, CD30, and the Epstein-Barr virus (EBV)-encoded latent membrane protein-1 (LMP1). Previous in situ hybridization studies have demonstrated TRAF1 transcripts in the malignant cells of the majority of Hodgkin's disease (HD) tumours, where the expression of TRAF1 was higher in EBV-associated tumours than in their EBV-negative counterparts. In order to determine whether TRAF1 and also TRAF2 were expressed at the protein level in HD and whether there was any relationship to EBV status, immunohistochemistry has been used to detect these proteins in a series of HD specimens. TRAF1 protein was detected more frequently in Hodgkin/Reed-Sternberg (HRS) cells from EBV-positive tumours than in their EBV-negative counterparts. This difference was statistically significant (p=0.01). In contrast, TRAF2 expression by HRS cells appeared to be independent of EBV status. Using a sequential labelling approach, co-localization of LMP1 with either TRAF1 or TRAF2 was also demonstrated in HRS cells from EBV-positive tumours. PMID- 11400144 TI - Ductal invasive G2 and G3 carcinomas of the breast are the end stages of at least two different lines of genetic evolution. AB - Ductal invasive grade (G) 2 and G3 carcinomas represent the majority of invasive breast cancers. Previous morphological and cytogenetic studies have provided evidence that ductal invasive G2 carcinoma may originate from at least two different genetic pathways. The aim of this study was to evaluate further the heterogeneity of G2 breast cancer in comparison with G3 cancers by cytogenetic and quantitative analysis. To this end, 35 cases of ductal invasive G2 and 42 cases of ductal invasive G3 carcinomas were investigated by means of comparative genomic hybridization (CGH) and these findings were correlated with DNA ploidy status, mitotic activity index (MAI), mean nuclear area (MNA), volume per lumen (VPL), and clinico-pathological parameters. The findings of this study demonstrate that ductal invasive G2 carcinomas, in contrast to ductal invasive G3 carcinomas, have to be interpreted as the morphological end stage resulting from two different cytogenetic and morphological pathways; the loss of 16q material is the cytogenetic key event in the evolution of a subgroup of this entity. By correlating genetic alterations with DNA ploidy status, an extended morphology based cytogenetic progression model is presented, with early and late genetic alterations in the pathogenesis of breast cancer. The correlation with MAI gives rise to the hypothesis that these different genetic pathways significantly differ in their proliferation rate. Further studies will be required to elucidate which genes contribute to an altered proliferation rate in these subgroups and to the associated prognosis. PMID- 11400145 TI - DNA image cytometry and human papillomavirus (HPV) detection help to select smears at high risk of high-grade cervical lesions. AB - Three samples were submitted from women undergoing routine screening (n=910): two smears (one for routine cytology and one for DNA image cytometry) and a scrape for human papillomavirus (HPV) testing. DNA histograms were classified as suspect in cases of aneuploidy, polyploidy, and/or diploidy with a high proliferation rate. Follow-up was available in 239 cases. The primary end-point was the presence of a high-grade squamous intraepithelial lesion (HGSIL) at biopsy. Seventy women (7.7%) had a high-risk (HR) HPV infection and a suspect DNA profile. In 77 women with cytological abnormalities, 28 HGSILs were detected: four with a prior diagnosis of ASCUS (all HR-HPV infected including three with a suspect DNA profile), three with smears evocative of LGSIL (all with HR-HPV infection and a suspect DNA profile), and 21 with smears evocative of HGSIL (all with HR-HPV infection and 20 with a suspect DNA profile). During the follow-up period, out of 239 women with a cytologically normal smear at first entry, five developed a HGSIL; all were HR-HPV-positive and four had a suspect DNA profile at the first smear. HR-HPV detection alone gives a sensitivity of 100% for the detection of HGSIL, with a specificity of 84.3%, whereas DNA measurement associated with HPV testing significantly enhances the specificity to 95.4%. Thus, the combination of HPV testing and DNA measurement provides a highly sensitive and specific evaluation of the risk of HGSIL on cervical smears. PMID- 11400146 TI - New chromosomal regions with high-level amplifications in squamous cell carcinomas of the larynx and pharynx, identified by comparative genomic hybridization. AB - Squamous cell carcinomas of the head and neck generally exhibit complex karyotypes. To gain better knowledge of the changes in the subgroup of laryngeal and pharyngeal squamous cell carcinoma, chromosomal gains and losses were investigated in 42 predominantly late-stage tumours, using comparative genomic hybridization. On average, 11.2 gains and 6.8 losses were found. Gains were detected in high frequencies at 1q, 3q, 5p, 7q, 8q, 11q13, 17q, and 18p, and losses at 3p, 4p, 5q, 11qter, and 18q. Neither the number nor the type of abnormalities, nor the occurrence of specific chromosome changes, was found to be related to DNA ploidy, tumour stage, or degree of differentiation. Apart from low level gains, many high-level amplifications were identified, in particular 3q24 qter (15 cases). Other regions recurrently involved were 11q13 (7 cases), 18p (5 cases), 18q11.2 (4 cases), and 8q23-24 and 11q14-22 (3 cases). Many of these amplified regions have not been reported before. Over half of all loci harbour genes coding for growth factors and growth factor receptors, suggesting an important role for such genes in squamous cell tumourigenesis and in the progression of late-stage tumours. PMID- 11400147 TI - Telomerase activity and telomere length in thyroid neoplasia: biological and clinical implications. AB - Despite several recent studies, the biological status and clinical relevance of telomerase expression in tumours derived from the thyroid follicular cell remain controversial. This study has analysed a series of normal, benign, and malignant thyroid samples using two novel approaches: the use of purified epithelial cell fractions to eliminate false-positives due to telomerase-positive infiltrating lymphocytes; and the simultaneous measurement of telomere length to provide a clearer interpretation of telomere dynamics in thyroid neoplasia. The data obtained support the prediction that the epithelial component of non-neoplastic thyroid and of follicular adenomas is telomerase-negative, any positive results being explicable by lymphocyte infiltration. In contrast, many malignant tumours, both follicular and papillary, were telomerase-positive. However, serial dilution of extracts indicated a wide spectrum of activity in these cancers, possibly related to variation in the proportion of telomerase-positive cells. Furthermore, an unexpectedly high proportion were telomerase-negative, a finding which was not explicable by technical problems such as TRAP (telomeric repeat amplification protocol) assay sensitivity. Many of these apparently telomerase-negative tumours had abnormally long telomeres. Correlation of telomerase and telomere length data suggests that thyroid cancers fall into three biological groups: telomerase positive lesions, consistent with the conventional model of telomere erosion followed by telomerase reactivation; telomerase-negative tumours, which maintain telomere length by a mechanism independent of telomerase; and telomerase-negative tumours which are still undergoing telomere erosion and may therefore be composed of mortal cancer cells. From a clinical standpoint, it is concluded that telomerase detection on unfractionated tissue, such as fine needle aspirates, is of no value as a marker of malignancy in follicular lesions, due to both low sensitivity and specificity. PMID- 11400148 TI - Inducible nitric oxide synthase expression in benign and malignant cutaneous melanocytic lesions. AB - Nitric oxide (NO) is synthesized by nitric oxide synthases (NOS) and plays an important role in tumour growth. In this study, inducible NOS (iNOS) expression was evaluated by immunohistochemistry in 34 melanocytic naevi (13 common melanocytic naevi, six Spitz naevi, and 15 so-called 'dysplastic naevi'), ten cutaneous melanomas in situ, 50 stage I invasive melanomas, and eight subcutaneous metastases of melanoma. In addition, four samples of melanocytic naevi and four samples of invasive melanomas were collected in order to perform western blot and northern blot analysis. By immunohistochemistry, melanocytic naevi never expressed iNOS. Among cases of melanoma in situ, two were negative, seven displayed staining in less than 20% of melanoma cells, and positivity was observed in 21-50% of melanoma cells in only one case. iNOS expression was detected in 46 out of 50 invasive melanomas (92%). Among these cases, 18 showed positivity in less than 20% of melanoma cells, 18 showed positivity in 21-50% of melanoma cells, and ten showed iNOS expression in more than 50% of cells. Statistical analysis revealed a significant difference in iNOS expression between melanocytic naevi and cutaneous melanomas (p<0.001). In addition, iNOS expression was significantly higher in invasive melanomas than in melanomas in situ (p=0.01). Among primary cutaneous melanomas, no significant correlation was found between iNOS expression and histopathological parameters (histotype, level, thickness and presence of regression/inflammatory infiltrate) and disease specific survival. In subcutaneous melanoma metastases, iNOS expression was diffuse in more than 50% of cells. Statistical analysis revealed that subcutaneous melanoma metastases showed greater iNOS immunoreactivity than invasive melanomas (p=0.02). Molecular analyses confirmed that iNOS mRNA and protein were highly expressed in melanoma samples. In conclusion, iNOS was constantly absent in melanocytic naevi, whereas it was frequently expressed in melanomas, with up-regulation of the enzyme paralleling tumour progression. These data suggest that iNOS may play a role in the malignant transformation of melanocytes and in tumour growth. In addition, iNOS may be useful as an immunohistochemical marker for malignant melanocytic lesions. PMID- 11400149 TI - MMP-9 is predominantly expressed in epithelioid and not spindle cell uveal melanoma. AB - Extracellular matrix-degrading enzymes are crucial for cancer metastases. One group of enzymes that has been increasingly implicated in the breakdown of the extracellular matrix, and hence the intravasation and dissemination of tumour cells, is the family of metalloproteinases. In the recent past, increasing efforts have led to the development of more or less specific matrix metalloproteinase (MMP) inhibitors. Data concerning the molecular nature and timing of the contribution of MMPs to tumour spread is of paramount importance in clarifying which MMP is an appropriate target for more selective MMP inhibition in future tumour therapy. This study immunohistochemically characterized the expression pattern of MMP-2, -3, and -9 in 26 uveal melanomas. Forty-six per cent of the uveal melanomas expressed MMP-2 and/or MMP-9. MMP-3 expression was seen in 17 out of 26 uveal melanomas. MMP-9, previously shown to play an important part in tumour dissemination, was predominantly present in epithelioid melanomas (71.4%) or the epithelioid portion of mixed cell uveal melanomas (67%), whereas only one out of ten spindle cell melanomas showed MMP-9 expression (10%). MMP-2 and MMP-9 expression was associated with a significantly higher incidence of metastatic disease. The survival rate of patients with MMP-2-positive melanomas was 31% vs. 85% for patients with MMP-2-negative (p<0.05); for MMP-9-positive uveal melanomas the survival rate was 27% vs. 85% with MMP-9-negative uveal melanomas (p<0.04). The fact that patients suffering from TIMP-1- as well as TIMP 2-positive uveal melanomas tended to show a better survival rate (72% vs. 45% for TIMP-1; 88% vs. 37% for TIMP-2) supports the view that proteolytic enzymes are of importance in tumour spread. PMID- 11400150 TI - Keratinocyte-derived growth factors play a role in the formation of hypertrophic scars. AB - In predisposed individuals, wound healing can lead to hypertrophic scar or keloid formation, characterized by an overabundant extracellular matrix. It has recently been shown that hypertrophic scars are accompanied by abnormal keratinocyte differentiation and proliferation, and significantly increased acanthosis, compared with normal scars. This study addressed the question of whether the development of normal and hypertrophic scars is regulated by differences in the growth factor profiles of both the epidermis and the dermis. The presence of interleukin-1alpha (IL-1alpha), IL-1beta, tumour necrosis factor-alpha (TNF alpha), platelet-derived growth factor (PDGF), transforming growth factor-beta1 (TGF-beta1), and basic fibroblast growth factor (bFGF) was investigated in biopsies taken from breast reduction scars at 3 and 12 months following surgery. The samples were analysed by immunohistological methods and categorized as scars that remained hypertrophic (HH), became normal (HN) or remained normal after 12 months (NN). The epidermal expression of IL-1alpha was significantly increased in NN scars compared with HN and HH scars 3 and 12 months following operation, whereas the dermal expression showed no difference. PDGF was significantly increased in the dermis of normal scars after 3 months and in both the epidermis and the dermis of hypertrophic scars after 12 months. IL-1beta, TNF-alpha, TGF beta and bFGF showed no differences. It is hypothesized that impaired production of keratinocyte-derived growth factors, such as IL-1alpha, leads to a decrease in the catabolism of the dermal matrix, whereas augmented epidermal PDGF production leads to increased formation of the dermal matrix in hypertrophic scars. These observations support the possibility that the epidermis is involved in preventing the formation of hypertrophic scars. PMID- 11400151 TI - Expression and induction of collagenases (MMP-8 and -13) in plasma cells associated with bone-destructive lesions. AB - Matrix metalloproteinases (MMPs) collectively degrade extracellular matrix and basement membrane proteins in chronic inflammation and bone-destructive lesions. This study examined the ability of immunoglobulin-producing plasma cells, typically present in sites of chronic inflammation, to express collagenases (MMP 8 and -13) in vivo and in vitro. Phorbol-12-myristate-13-acetate, interleukin-6, and tumour necrosis factor-alpha and heparin with the tumour promoter or cytokines potently enhanced (up to nine-fold) MMP-8 and -13 expression by the RPMI 8226 myeloma cell line, as evidenced by western blotting and semi quantitative reverse transcriptase-polymerase chain reaction. Immunohistochemical analysis and in situ hybridization revealed that plasma cells expressed MMP-8 and -13 focally in periapical granulomas, odontogenic cysts, and malignant plasmacytomas. MMP-8 and MMP-13 from plasma cells can participate in bone organic matrix destruction at sites of chronic inflammation and neoplastic growth. Since MMP-13 was more frequently expressed than MMP-8 in plasma cells of strongly recurring keratocysts and malignant plasmacytomas, it is concluded that plasma cell MMP-13 has a particularly important role in benign and malignant bone destructive lesions. PMID- 11400152 TI - Expression of MMP2, MMP9, MT1-MMP, TIMP1, and TIMP2 mRNA in valvular lesions of the heart. AB - Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play an important role in several diseases. This study was undertaken to investigate the mRNA synthesis of MMP2, MMP9, membrane-type 1 (MT1)-MMP, and matrix metalloproteinase inhibitors TIMP1 and TIMP2 by in situ hybridization in a set of heart mitral and aortic valves operatively removed due to degenerative or inflammatory valvular diseases. The material consisted of 21 valves, eight with endocarditis and 13 with a degenerative valvular disease. The samples were studied by in situ hybridization with specific probes for MMP2, MMP9, MT1-MMP, TIMP1, and TIMP2. Synthesis of MMP2 mRNA was found in seven valves, five with endocarditis and two with degenerative valvular disease. Signals for MMP9 mRNA were found in two cases with endocarditis and five cases with degenerative valvular disease. No signal for MT1-MMP mRNA was found in the lesions. TIMP1 mRNA, on the other hand, was found in 17 cases, both endocarditis and degenerative valvular disease. TIMP2 mRNA was found in three cases of endocarditis. The signals for MMP2, MMP9, TIMP1, and TIMP2 mRNA were localized in endothelial cells and in fibroblast-like cells expressing alpha-smooth muscle actin, thus showing myofibroblast-type differentiation. The results show that matrix metalloproteinases MMP2 and MMP9, and matrix metalloproteinase inhibitors TIMP1 and TIMP2 mRNAs are synthesized in diseased valves and suggest that they may contribute to matrix remodelling in valvular disease. PMID- 11400153 TI - In vivo collagenase-2 (MMP-8) expression by human bronchial epithelial cells and monocytes/macrophages in bronchiectasis. AB - The purpose of this study was to determine whether other cellular sources than neutrophils can express matrix metalloproteinase (MMP)-8 protein and mRNA in bronchiectatic (BE) lung. The molecular forms of MMP-8 in the BE bronchoalveolar lavage fluid (BALF) and healthy control BALF were analysed by western immunoblotting. MMP-8 expression was demonstrated by immunohistochemistry and in situ hybridization in BE lung tissue and by immunohistochemistry in control lung tissue. In the BE BALF, different MMP-8 species were detected: 70-80 kD MMP-8 apparently of polymorphonuclear leukocyte (PMN) origin and also 40-60 kD MMP-8 from non-PMN cellular sources, such as bronchial epithelial cells, glandular cells or monocytes/macrophages. Both of these MMP-8 species were elevated and converted to a significant extent to activated forms in BE BALF compared with healthy control BALF. The levels of high molecular weight (>80 kD) MMP-8 complexes, evidently representing MMP-8 trapped by endogenous MMP inhibitors and/or MMP-8 dimers, were significantly elevated in BE BALF compared with healthy control BALF. In BE lung tissue, the MMP-8 protein and mRNA expression was found in bronchial ciliated epithelial cells, glandular cells, neutrophils, and monocytes/macrophages infiltrating the bronchial epithelial area. Minimal MMP-8 expression was observed in neutrophils, monocytes/macrophages, and epithelial cells in control lung tissues. In this study, new potential cellular sources have been demonstrated for MMP-8 in the inflamed lung. MMP-8 from multiple cellular sources, including inflamed lung epithelium, was activated to a significant extent in the BE BALF, indicating a major role for MMP-8 in the destruction of lung and bronchial tissues. PMID- 11400154 TI - Potential role for monocyte chemotactic protein-4 (MCP-4) in monocyte/macrophage recruitment in acute renal inflammation. AB - The CC chemokine, monocyte chemoattractant protein-4 (MCP-4), is an important chemoattractant for monocytes and T cells. Recent data indicate a role in renal inflammation. This study has used in situ hybridization and immunohistochemical analysis of cryostat sections of biopsy material taken from patients with acute renal allograft rejection and vasculitic glomerulonephritis to demonstrate renal expression of MCP-4, both at message and protein level. MCP-4 was primarily expressed at peritubular, periglomerular, and perivascular sites, irrespective of the inflammatory condition, and was associated with infiltrating CD3-positive lymphocytes and CD68-positive monocyte/macrophages. In addition, proximal tubular epithelial cells grown in culture from cortical fragments of human kidney showed low levels of constitutive MCP-4 expression, detectable by western blotting; this expression of MCP-4 was up-regulated in response to the pro-inflammatory cytokines, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN gamma). CCR3-, CCR5- and CCR2-expressing leukocyte populations were identified at sites of MCP-4 expression. Double-staining techniques revealed that CC chemokine receptor-expressing cells were primarily CD68-positive. These studies suggest an important role for MCP-4 in the recruitment and retention of monocytes/macrophages in renal inflammation. PMID- 11400155 TI - Nerve growth factor and brain-derived neurotrophic factor mRNAs are regulated in distinct cell populations of rat heart after ischaemia and reperfusion. AB - Neurotrophins play a crucial role in the development of the peripheral nervous system and their mRNAs are often regulated after several types of tissue injury. This study has investigated the regulation of nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), and neurotrophin-3 (NT-3) mRNAs 30 min after myocardial ischaemia followed by reperfusion, by northern blotting, and in situ hybridization in a rat model. Between 2 and 120 h of reperfusion, Ngf mRNA levels showed two- to four-fold up-regulation compared with sham-operated hearts. Scattered Ngf-expressing cells, probably pericytes, were detected in the viable border zone of the myocardium in close association with capillaries, venules, and arterioles. In addition, diffuse Ngf expression was seen in the infarct area after 120 h of reperfusion. Bdnf mRNA showed transient up-regulation after 2 and 5 h of reperfusion and remained at control levels thereafter. Bdnf was expressed in the myocytes of the viable border zone. Nt-3 expression showed no significant changes compared with sham-operated hearts. These results suggest a role for NGF and/or BDNF in the pathogenesis of reperfusion injury or in the alterations of cardiac sensory and sympathetic neuronal function after myocardial ischaemia and reperfusion. PMID- 11400156 TI - Expression of CD31 by cells of extensive ductal in situ and invasive carcinomas of the breast. AB - CD31, an adhesion molecule expressed by endothelial cells, leukocytes, and platelets, is used in surgical pathology as a marker of normal and neoplastic vascularization. During the assessment of angiogenesis in breast carcinomas, CD31 expression was observed in a single case of large (5.2 cm diameter) high nuclear grade ductal carcinoma in situ (HG-DCIS) associated with poorly differentiated invasive ductal carcinoma (G3-IDC). Expression was limited to the cell membrane. This study focused on 32 HG-DCIS> or = 2 cm, either pure or associated with IDC. Cancer cells wereCD31(+) in 11 cases. Double staining using anti-CD31 monoclonal antibody (MAb) and anti-CD44 MAb, the anti-hyaluronate receptor, showed that foci of CD31(+) and CD44(-) tumour cells could be traced throughout the glandular tree, marking the intraductal diffusion of tumour up to Paget's cells at the nipple. The associated G3-IDC and their lymph node metastases were instead CD31(+) and CD44(+). CD31(+) tumours were oestrogen receptor (ER)(-), frequently p53(+) and c-erb-B2(+), and infiltrated by CD4(+) T lymphocytes. Normal and hyperplastic epithelia were constantly CD31(-). Other endothelial markers (e.g Factor VIII-RA and CD34) were not expressed by carcinoma cells, as was CD38, the CD31 ligand. In conclusion, CD31 expression is a feature acquired by breast cancer cells in the DCIS model. CD31 expression mainly correlates with tumour cells spreading within the ductal system. Finally, the invasive phenotype requires the co-expression of CD31 and CD44. PMID- 11400157 TI - Overproduction of perlecan core protein in cultured cells and transgenic mice. AB - Heparan sulphate proteoglycan (HSPG) and amyloid P component are the only macromolecules consistently associated with all varieties of amyloid, irrespective of the type of amyloid protein, suggesting that HSPG may play a pathogenetic role in amyloid formation through a common mechanism. In the case of Alzheimer's disease (AD), HSPG, such as perlecan, co-accumulates with amyloid beta protein (Abeta), a main constituent of amyloid plaques, and paired helical filaments (PHFs). Additionally, in vitro, HSPG accelerates both Abeta fibril and PHF formation and protects Abeta from degradation. Therefore, this study first established lines of P19 mouse embryonic carcinoma cells stably carrying an expression vector encoding the complete perlecan core protein (approximately 400 kD). In the cell lysates, overexpressed perlecan was identified as an approximately 400 kD protein without glycosaminoglycan side-chains, while in the media, secreted perlecan was mostly glycosylated, suggesting that the secretion and glycosylation of perlecan are coupled. Next, transgenic mice were produced using the same expression vector. Marked perlecan overexpression occurred in the cytoplasm of multiple tissues including the brain, heart, kidney, and pancreas, without a discernible increase of perlecan in extracellular matrices. The transgenic mice up to 18 months of age did not develop amyloid or AD-like pathology in the brain or elsewhere, based on histochemical and immunohistochemical analyses. Thus, overproduction of perlecan core protein is insufficient to lead to amyloidosis and AD-like pathology. PMID- 11400158 TI - Characterization of tissue-specific LIM domain protein (FHL1C) which is an alternatively spliced isoform of a human LIM-only protein (FHL1). AB - We have cloned and characterized another alternatively spliced isoform of the human four-and-a-half LIM domain protein 1 (FHL1), designated FHL1C. FHL1C contains a single zinc finger and two tandem repeats of LIM domains at the N terminus followed by a putative RBP-J binding region at the C-terminus. FHL1C shares the same N-terminal two-and-a-half LIM domains with FHL1 but different C terminal protein sequences. Due to the absence of the exon 4 in FHL1C, there is a frame-shift in the 3' coding region. Sequence analysis indicated that FHL1C is the human homolog of murine KyoT2. The Northern blot and RT-PCR results revealed that FHL1 is widely expressed in human tissues, including skeletal muscle and heart at a high level, albeit as a relatively low abundance transcript in brain, placenta, lung, liver, kidney, pancreas, and testis. In contrast, FHL1C is specifically expressed in testis, skeletal muscle, and heart at a relatively low level compared with FHL1. The expression of FHL1C transcripts was also seen in aorta, left atrium, left, and right ventricles of human heart at low level. Immunoblot analysis using affinity-purified anti-FHL1C antipeptide antibodies confirmed a 20 kDa protein of FHL1C in human skeletal muscle and heart. Unlike FHL1B, which is another FHL1 isoform recently reported by our group and localized predominantly in the nucleus [Lee et al., 1999], FHL1C is localized both in the nucleus and cytoplasm of mammalian cell. PMID- 11400159 TI - Adenovirus-mediated BMP2 expression in human bone marrow stromal cells. AB - Recombinant adenoviral vectors have been shown to be potential new tools for a variety of musculoskeletal defects. Much emphasis in the field of orthopedic research has been placed on developing systems for the production of bone. This study aims to determine the necessary conditions for sustained production of high levels of active bone morphogenetic protein 2 (BMP2) using a recombinant adenovirus type 5 (Ad5BMP2) capable of eliciting BMP2 synthesis upon infection and to evaluate the consequences for osteoprogenitor cells. The results indicate that high levels (144 ng/ml) of BMP2 can be produced in non-osteoprogenitor cells (A549 cell line) by this method and the resultant protein appears to be three times more biologically active than the recombinant protein. Surprisingly, similar levels of BMP2 expression could not be achieved after transduction with Ad5BMP2 of either human bone marrow stromal cells or the mouse bone marrow stromal cell line W20-17. However, human bone marrow stromal cells cultured with 1 microM dexamethasone for four days, or further stimulated to become osteoblast like cells with 50 microg/ml ascorbic acid, produced high levels of BMP2 upon Ad5BMP2 infection as compared to the undifferentiated cells. The increased production of BMP2 in adenovirus transduced cells following exposure to 1 microM dexamethasone was reduced if the cells were not given 50 microg/ml ascorbic acid. When bone marrow stromal cells were allowed to become confluent in culture prior to differentiation, BMP2 production in response to Ad5BMP2 infection was lost entirely. Furthermore, the increase in BMP2 synthesis seen during differentiation was greatly decreased when Ad5BMP2 was administered prior to dexamethasone treatment. In short, the efficiency of adenovirus mediated expression of BMP2 in bone marrow stromal cells appears to be dependent on the differentiation state of these cells. PMID- 11400160 TI - Cactus-independent nuclear translocation of Drosophila RELISH. AB - Insects can effectively and rapidly clear microbial infections by a variety of innate immune responses including the production of antimicrobial peptides. Induction of these antimicrobial peptides in Drosophila has been well established to involve NF-kappaB elements. We present evidence here for a molecular mechanism of Lipopolysaccharide (LPS)-induced signaling involving Drosophila NF-kappaB, RELISH, in Drosophila S2 cells. We demonstrate that LPS induces a rapid processing event within the RELISH protein releasing the C-terminal ankyrin repeats from the N-terminal Rel homology domain (RHD). Examination of the cellular localization of RELISH reveals that the timing of this processing coincides with the nuclear translocation of the RHD and the retention of the ankyrin-repeats within the cytoplasm. Both the processing and the nuclear translocation immediately precede the expression of antibacterial peptide genes cecropin A1, attacin, and diptericin. Over-expression of the RHD but not full length RELISH results in an increase in the promoter activity of the cecropin A1 gene in the absence of LPS. Furthermore, the LPS-induced expression of these antibacterial peptides is greatly reduced when RELISH expression is depleted via RNA-mediated interference. In addition, loss of cactus expression via RNAi revealed that RELISH activation and nuclear translocation is not dependent on the presence of cactus. Taken together, these results suggest that this signaling mechanism involving the processing of RELISH followed by nuclear translocation of the RHD is central to the induction of at least part of the antimicrobial response in Drosophila, and is largely independent of cactus regulation. PMID- 11400161 TI - p53 down-regulates human bradykinin B1 receptor gene expression. AB - The tumor suppressor, p53, has been shown to transcriptionally activate or silence a number of target genes. As an activator, p53 relies on its specific consensus sequence within the promoter. It is not clear whether p53 requires a specific DNA binding site in its action as a gene repressor. This report demonstrates that the human BKB1R gene is a p53 target. Expression of p53 in transiently transfected SV40-transformed IMR90 cells strongly suppressed luciferase reporter activity driven by a 1.8 kb BKB1R promoter as well as its minigene. These down-regulations were p53 dose-dependent. p53 reduced both basal and induced promoter activities of the minigene. Expression of p53 abolished the inducibility of the minigene. Induction of endogenous p53 expression by etoposide also inhibited promoter activity and minigene inducibility. Replacing the region containing both the putative p53 binding site and the TATA-box with a basal adenovirus promoter in the 1.8 kb promoter construct did not prevent p53 from inhibiting BKB1R promoter activity. Thus suppression by p53 is not mediated by competition with the TATA-binding protein and is not through interaction with the putative p53-binding site. p53 also does not appear to suppress BKB1R gene expression through interaction with c-Jun which functions in the inducibility of this gene [Yang et al., 2001]. PMID- 11400162 TI - Fibromodulin is expressed by both chondrocytes and osteoblasts during fetal bone development. AB - Fibromodulin, a keratan-sulfate proteoglycan, was first isolated in articular cartilage and tendons. We have identified fibromodulin as a gene regulated during BMP-2-induced differentiation of a mouse prechondroblastic cell line. Because expression of fibromodulin during endochondral bone formation has not been studied, we examined whether selected cells of the chondrocytic and osteoblastic lineage expressed fibromodulin. Fibromodulin mRNA was detected in conditionally immortalized murine bone marrow stromal cells, osteoblasts, and growth plate chondrocytes, as well as in primary murine calvarial osteoblasts. We, therefore, investigated the temporo-spatial expression of fibromodulin in vivo during endochondral bone formation by in situ hybridization. Fibromodulin was first detected at 15.5 days post coitus (dpc) in the perichondrium and proliferating chondrocytes. Fibromodulin mRNA was also detected at 15.5 dpc in the bone collar and periosteum. At later time points fibromodulin was expressed in the primary spongiosa and the endosteum. To determine whether fibromodulin was expressed during intramembranous bone formation as well, in situ hybridization was performed on calvariae. Fibromodulin mRNA was present in calvarial osteoblasts from 15.5 dpc. These results demonstrate that fibromodulin is developmentally expressed in cartilage and bone cells during endochondral and intramembranous ossification. These findings suggest that this extracellular matrix protein plays a role in both endochondral and intramembranous bone formation. PMID- 11400163 TI - Stimulation of hyaluronan synthesis by interleukin-1beta involves activation of protein kinase C betaII in fibroblasts from patients with Graves' ophthalmopathy. AB - Hyaluronan accumulation in the retroorbital connective tissue is one of the pathological features of Graves' ophthalmopathy. Interleukin-1beta (IL-1beta) is known to stimulate hyaluronan synthesis in orbital fibroblasts. In the present study, the intracellular signal transduction pathways involved in this stimulatory effect were investigated in cultured human retroorbital fibroblasts from patients with Graves' ophthalmopathy. IL-1beta-induced hyaluronan synthesis was significantly inhibited by pretreatment of the cells with two protein kinase C (PKC) inhibitors, chlerythrine chloride and H-7. In addition, treatment with phorbol 12-myristate 13-acetate (PMA), a direct PKC activator, also resulted in increased hyaluronan production. IL-1beta- or PMA-stimulated hyaluronan synthesis was blocked by the protein synthesis inhibitor, cycloheximide. Moreover, the intracellular Ca(2+) concentration of the orbital fibroblasts was also involved in the IL-1beta induced transduction pathway, the effect being completely inhibited by BAPTA, an internal calcium chelator. In addition, A23187, a calcium ionophore, increased hyaluronan synthesis in unstimulated cells. These results suggest that the Ca(2+)-dependent PKC signal transduction pathway plays an important role in the IL-1beta-induced hyaluronan synthesis. Moreover, IL-1beta treatment resulted in increased PKC activity and the rapid translocation of PKC betaII from the cytoplasm to the plasma membrane. These results indicate that cytosolic Ca(2+) and PKC betaII are involved in IL-1beta-induced hyaluronan synthesis in cultured orbital fibroblasts from patients with Graves' ophthalmopathy. PMID- 11400164 TI - Inhibition of p38MAP kinase potentiates the JNK/SAPK pathway and AP-1 activity in monocytic but not in macrophage or granulocytic differentiation of HL60 cells. AB - Monocytic differentiation of HL60 cells induced by 1,25-dihydroxyvitamin D(3) (1,25 D(3)) has been recently shown (Exp Cell Res 258, 425, 2000) to be enhanced by an exposure to SB203580 or to SB202190, specific inhibitors of p38MAP kinase, with concomitant up-regulation of the c-jun N terminal kinase (JNK) pathway. In the present study we inquired if this enhancement and the JNK up-regulation are limited to 1,25 D(3)-induced differentiation, or if they occur more generally in HL60 cell differentiation. We found that dimethylsulfoxide (DMSO)-induced differentiation, and to a lesser extent tetradecanoylphorbol acetate (TPA) induced macrophage differentiation were also potentiated by the p38MAPK inhibitors, but that granulocytic differentiation in response to all-trans retinoic acid (RA) was not. The enhancement of differentiation by p38MAPK inhibitors was accompanied by an activation of the JNK MAPK pathway, as shown by the phosphorylation levels of these kinases and by AP-1 binding, but only in 1,25 D(3)-treated cells. This shows that an up-regulation of the JNK stress pathway during 1,25 D(3)-induced monocytic differentiation occurs selectively in this lineage of differentiation and is not necessary for the expression of the differentiated phenotype. PMID- 11400165 TI - Different effects of genistein on molecular markers related to apoptosis in two phenotypically dissimilar breast cancer cell lines. AB - The association between consumption of genistein-containing soybean products and lower risk of breast cancer suggests a cancer chemopreventive role for genistein. Consistent with this suggestion, exposing cultured human breast cancer cells to genistein inhibits cell proliferation, although this is not completely understood. To better understand how genistein works, the ability of genistein to induce apoptosis was compared in phenotypically dissimilar MCF-7 and MDA-MB-231 human breast cancer cells that express the wild-type and mutant p53 gene, respectively. After 6 days of incubation with 50 microM genistein, MCF-7 but not MDA-MB-231 cells, showed morphological signs of apoptosis. Marginal proteolytic cleavage of poly-(ADP-ribose)-polymerase and significant DNA fragmentation were also detected in MCF-7 cells. In elucidating these findings, it was determined that after 2 days of incubation with genistein, MCF-7 but not MDA-MB-231 cells, had significantly higher levels of p53. Accordingly, the expression of certain proteins modulated by p53 was studied next. Levels of p21 increased in both of the genistein-treated cell lines, suggesting that p21 gene expression was activated but in a p53-independent manner, whereas no significant changes in levels of the pro-apoptotic protein, Bax, were found. In MCF-7 cells, levels of the anti-apoptotic protein, Bcl-2, decreased slightly at 18-24 h but then increased considerably after 48 h. Hence, the Bax:Bcl-2 ratio initially increased but later decreased. These data suggest that at the genistein concentration tested, MCF-7 cells in contrast to MDA-MB-231 cells were sensitive to the induction of apoptosis by genistein, but Bax and Bcl-2 did not play clear roles. PMID- 11400166 TI - Calcium oscillations in Xenopus egg cycling extracts. AB - Cell cycle in various types of cells and in early embryos is often accompanied by transient changes in the concentration of free cytosolic calcium. In the present study, using fluorescent indicator fura-2, we demonstrate that Ca(2+) oscillates cyclically with an amplitude of about 100 nM and a period of mitotic cycle in cell-free Xenopus egg cycling extracts. It peaks in early metaphase just preceding mitotic reactivation of Cdc2 kinase and MAPK and reaches a minimum in interphase. The source of Ca(2+) in the extracts is a particulate fraction containing egg intracellular Ca(2+) stores, since the addition of a calcium mobilizing second messenger, inositol 1,4,5-trisphosphate (IP3), induced a transient increase in Ca(2+). The inclusion of heparin, an IP3 receptor antagonist, or ultrafiltration of the extracts prevented Ca(2+)-releasing activity of IP3. The depletion of Ca(2+) in the extracts by the calcium chelator BAPTA resulted in the blockade of cell cycle at different stages, depending on the time of drug administration. The addition of BAPTA late in interphase blocked cell cycle at mitotic entry in prophase, whereas its application in anaphase or telophase blocked the extracts in early interphase. BAPTA administration in metaphase before transition to anaphase brought about a metaphase-like arrest in the cycling extracts. Inhibition of IP3-induced calcium release by heparin also arrested cell cycle progression in the cycling extracts. PMID- 11400167 TI - Human endothelial cells grow poorly on vitronectin: role of PAI-1. AB - The cell adhesive protein vitronectin is a common component of interstitial extracellular matrix and circulates in plasma. It competes effectively with other plasma proteins to adsorb to certain biomaterial surfaces, and is likely to represent an important cell adhesion mediator on the luminal surface of vascular grafts. It is also found associated with certain vascular pathologies. We have shown previously that human endothelial cells grow poorly on a vitronectin surface compared with other extracellular matrix molecules. In this paper we show that endothelial cells seeded on vitronectin and fibronectin produced substantially different profiles of extracellular matrix molecules. The most outstanding difference was in the amount of matrix-localised plasminogen activator-inhibitor-1 which was high on vitronectin and negligible on fibronectin. This was correlated with a small but significant inhibition of cell adhesion to vitronectin compared with fibronectin, and very significant interference with dissociation of cell: extracellular matrix contacts, resulting either from direct inhibition of the proteolytic activity of urokinase, or from interference with urokinase-receptor signaling and consequent focal adhesion turnover. Such interference would inhibit cell proliferation by disabling the cells from loosening their matrix contacts in order to proceed through mitosis. This would seriously compromise endothelial recovery in cases of damage to the vascular wall and placement of stents or grafts, where the presence of surface adsorbed vitronectin is likely to modulate the tissue response. PMID- 11400168 TI - 26S proteasome inhibition induces apoptosis and limits growth of human pancreatic cancer. AB - The 26S proteasome degrades proteins that regulate transcription factor activation, cell cycle progression, and apoptosis. In cancer, this may allow for uncontrolled cell division, promoting tumor growth, and spread. We examined whether selective inhibition of the 26S proteasome with PS-341, a dipeptide boronic acid analogue, would block proliferation and induce apoptosis in human pancreatic cancer. Proteasome inhibition significantly blocked mitogen (FCS) induced proliferation of BxPC3 human pancreatic cancer cells in vitro, while arresting cell cycle progression and inducing apoptosis by 24 h. Accumulation of p21(Cip1-Waf-1), a cyclin dependent kinase (CDK) inhibitor normally degraded by the 26S proteasome, occurred by 3 h and correlated with cell cycle arrest. When BxPC3 pancreatic cancer xenografts were established in athymic nu/nu mice, weekly administration of 1 mg/kg PS-341 significantly inhibited tumor growth. Both cellular apoptosis and p21(Cip1-Waf-1) protein levels were increased in PS-341 treated xenografts. Inhibition of tumor xenograft growth was greatest (89%) when PS-341 was combined with the tumoricidal agent CPT-11. Combined CPT-11/PS-341 therapy, but not single agent therapy, yielded highly apoptotic tumors, significantly inhibited tumor cell proliferation, and blocked NF-kappaB activation indicating this systemic therapy was effective at the cancer cell level. 26S proteasome inhibition may represent a new therapeutic approach against this highly resistant and lethal malignancy. PMID- 11400169 TI - Inactivation of multiple targets by nitric oxide in CD95-triggered apoptosis. AB - Nitric oxide (NO) plays an important anti-apoptotic role by inactivating both upstream and downstream apoptotic molecules. We now report that exogenously supplied NO protected Jurkat T cells from anti-CD95-stimulated apoptosis. We have recently shown that nitrosation of the activator protein-1 (AP-1) transcriptional factor is crucial for NO-mediated inhibition of cell death triggered by etoposide or ceramide. Since the inhibition of apoptosis by NO has been reported to involve AP-1, we evaluated its involvement in in CD95-mediated cell death. Cross-linking of CD95 enhanced AP-1 DNA binding activity and AP-1-dependent CD95L transactivation, which were both significantly reduced by different NO-donors compounds. However, AP-1 induction does not seem to significantly contribute to anti-CD95-triggered apoptosis, as cell death could not be prevented by using the recombinant Fas-Fc fusion protein which inhibits the CD95/CD95L interaction. We observed that caspase 3-like activity was negatively modulated by several NO donors in vitro and that titratable thiol groups of purified caspases 3, 7, and 9 decreased in the presence of NO-releasing compounds. In conclusion, we demonstrated that NO-mediated inhibition of other targets, possibly caspases, but not AP-1, is a crucial event responsible for protection against anti-CD95 stimulated apoptosis. Even though NO affects multiple molecular mechanisms, the relevant target for exerting the cellular effects, may vary among different models. PMID- 11400170 TI - Molecular identification of NMDA glutamate receptors expressed in bone cells. AB - The N-methyl-D-aspartate (NMDA) subtype of the glutamate receptor has recently been identified in bone, but the molecular composition of this receptor expressed by bone cells is unknown. NMDA receptor (NMDAR) is a hetero-oligomeric protein composed of two classes of subunits, the essential subunit NR1 and NR2A to D subunits that do not by themselves produce functional channels but potentiate NR1 activity and confer functional variability to the receptor. These subunits coassemble in different combinations to form functionally distinct NMDAR. In this study, we have investigated the molecular composition of NMDAR expressed by osteoblasts and osteoclasts in culture, using RT-PCR analysis, in situ hybridization and immunocytochemistry. Specific probes were designed for the different subunits of the NMDAR, and we showed by RT-PCR analysis that mammalian osteoclasts expressed NR2B and NR2D subunits mRNAs but not NR2A and NR2C mRNAs. Rat calvaria and MG63 osteoblastic cells also expressed several NR2 subunits mRNAs, namely NR2A, NR2B, and NR2D. In situ hybridization on isolated rabbit osteoclasts and MG63 cells has confirmed the localization of NR1, NR2B, and NR2D transcripts in osteoclasts and NR1, NR2A, NR2B, and NR2D transcripts in MG63 cells. The expression of NR2D protein by bone cells was shown by immunofluorescence. These results demonstrate for the first time that osteoblasts and osteoclasts express several NR2 subunits, suggesting a molecular diversity of NMDAR channels similar to what was shown for brain. The presence of distinct functional NMDAR on bone cells may be associated with various states of bone cell differentiation and function. PMID- 11400171 TI - Non-natural CBP2 binding peptides and peptomers modulate carcinoma cell adhesion and invasion. AB - A combinatorial approach that utilized a repertoire of bacteriophage-peptides has identified a number of non-natural CBP2 binding peptides. Moreover, co localization of some of these peptides with CBP2 in a number of tumor cell lines demonstrated that the peptides were directed to an intracellular location spatially coincident with the normal distribution of CBP2 [Sauk et al., 2000]. From among these sequences WHYPWFQNWAMA and LDSRYSLQAAMY were the most effective CBP2 binding peptides and best fulfilled the combinatorial motif containing deep hydrophobic pockets. When the hydropathic profiles of collagen alpha1(IV) and alpha2 (IV) were compared with these dodecapeptides, the hydropathic profiles of WHYPWFQNWAMA and LDSRYSLQAAMY closely matched those of alpha1(IV) 414-452 and alpha1(IV)531-543. These peptides were shown to be functional peptidomimics and possessed the ability to alter cell adhesion and invasion of human squamous cell carcinoma cell lines. Peptomers were formed of these non-natural peptides to explore the role that a repetitive peptide may have on cell adhesion. The enhanced cell adhesion observed with the peptomers required both CBP2 antibodies and integrin antibodies for inhibition. The enhanced adhesion observed even in the face of combined antibody inhibition was consistent with such complexes possessing correspondingly slower dissociation rates. Thus, suggesting that peptomers may function in a like manner to multimeric peptide MHC complexes (tetramers) binding more than one cell receptor on a specific cell. These findings evoke both peptidomimics of native ligands and their peptomers as potential reagents by which to target tumor cells for chemotherapy, imaging, or retargeting viral vectors for gene therapy. PMID- 11400172 TI - Human T cell responses to endogenously presented HLA-A*0201 restricted peptides of Simian virus 40 large T antigen. AB - Presence of the simian virus 40 (SV40) has recently been demonstrated in a relatively high percentage of human mesotheliomas and it is associated with the development of these malignancies in pleural cavities. Therefore, we have initiated a study to identify candidate peptides presented by the human HLA A*0201 molecule for vaccination approaches against SV40 and monitoring of SV40 directed human immune responses. Initial screening of SV40 large T (Tag) domains required for transformation of cells for HLA-A*0201 binding motifs revealed ten possible binding peptides. Screening of these candidate peptides showed that seven of the ten peptides could bind and stabilize HLA-A*0201 molecules. In an in vitro immunization assay the two peptides with the highest binding affinity for HLA-A*0201, Tag aa 396-405 and aa 577-585, were tested for their ability to induce peptide specific cytotoxic T cells in two healthy donors. One donor developed cytotoxic T cells against Tag aa 396-405 and in T cell cultures of both donors Tag aa 577-585 specific T cells were initiated. The T cells against Tag aa 577-585 not only recognized and killed peptide pulsed cells, but, most importantly, SV40 transformed human mesothelial cells. This is the first demonstration of the induction of SV40 specific human cytotoxic T lymphocytes that recognize endogenously processed peptides from SV40. This peptide identification study opens the possibility to investigate immune responses against SV40 in mesothelioma patients and in individuals exposed to SV40. PMID- 11400173 TI - Mediator caused induction of a human bradykinin B1 receptor minigene: participation of c-Jun in the process. AB - The bradykinin B1 receptor (BKB1R) gene is expressed in selected tissues such as lung and kidney. In these tissues it is expressed at a very low level until induced by inflammatory mediators. Our aim has been to understand the mechanism of this regulatory process. A human BKB1R minigene was constructed. It contained a 1.8 kb promoter, the entire exon I, 1.5 kb of intron I, the entire exon II and intron II, and the luciferase gene as a reporter. Transient transfection of the minigene into SV40-transformed IMR90 cells (IMRSV) resulted in a promoter activity which was activated by the mediators, lipopolysaccharide and (LPS) desArg(10)-kallidin. In contrast, these mediators did not induce the activity of the 1.8 kb promoter construct alone. Thus, motifs exclusive of the promoter such as 5'-UTR and/or intron regions are required for mediator-induced expression of this gene. Promoter activities of both the minigene and the 1.8 kb promoter construct were enhanced in a dose-dependent manner upon cotransfection with c Jun. Furthermore, cotransfecting c-Jun with the minigene achieved the maximal promoter activity with no further increase in response to mediators. Conversely, the induction of the minigene promoter activity by mediators was abolished upon cotransfection with a dominant negative mutant of c-Jun. Other experiments suggest that multiple AP-1 sites are interactive with the c-Jun upregulation of this gene. Taken together, these results point to c-Jun as a key intermediary in the activation of the expression of this gene by mediators. However, participation of motifs outside of the promoter are necessary to obtain this inducible expression. PMID- 11400174 TI - Serum suppresses myeloid progenitor apoptosis by regulating iron homeostasis. AB - The growth and survival of committed hematopoietic progenitors is dependent upon cytokine signaling. However, serum is also required for optimal growth of these progenitors in culture ex vivo. Here we report that serum withdrawal leads to myeloid progenitor cell apoptosis. Although serum deprivation-induced cell death has many hallmarks typical of apoptosis, these cell deaths were not inhibited by hemopoietins, survival factors such as IGF-I, or treatment with a broad-spectrum caspase inhibitor. Rather, apoptosis due to serum withdrawal was associated with damage to mitochondria. Surprisingly the serum factor required for myeloid cell survival was identified as iron, and loss of iron led to marked reductions in ATP production. Furthermore, supplementing serum-deprived myeloid cells with bound or free iron promoted cell survival and prevented mitochondrial damage. Therefore, serum suppresses hematopoietic cell apoptosis by providing an obligate source of iron and iron homeostasis is critical for proper myeloid cell metabolism and survival. PMID- 11400175 TI - Effect of endogenous dopamine on extrastriatal [11C]FLB 457 binding measured by PET. AB - Central dopaminergic systems are known to be implicated in the pathophysiology of schizophrenia and recent in vivo dopamine receptor imaging studies have focused on the measurement of extrastriatal dopamine receptor. However, there are only a limited number of ligands that can measure the low-density D2 receptor in extrastriatal regions and their sensitivity to endogenous dopamine in extrastriatal regions has not yet been fully examined. In this study, the effect of endogenous dopamine on the extrastriatal binding of [11C]FLB 457 was examined in the rhesus monkey after facilitation with 1 mg/kg of methamphetamine (MAP) and was compared with the effect on the striatal binding of [11C]raclopride. The indices of receptor binding were obtained by four methods using cerebellum as a reference region. The bindings of [11C]FLB 457 in the frontal cortex, temporal cortex, and thalamus were not significantly changed after MAP treatment, while the striatal binding of [11C]raclopride was decreased by more than 20%. These results suggest that [11C]FLB 457 is not sensitive to endogenous dopamine in the extrastriatal regions of rhesus monkeys, despite a sufficient dose of MAP to decrease the binding of [11C]raclopride in the striatum. PMID- 11400176 TI - Quantitative ultrastructure of slowly adapting lingual afferent terminals in the principal and oral nuclei in the cat. AB - Previous studies provide evidence that a structure/function correlation exists in the cytoarchitectonically different zones of the trigeminal sensory nuclei. To extend this relationship, we examined the ultrastructural features of trigeminal primary afferent neurons in the cat dorsal principal nucleus (Vpd) and the rostrodorsomedial oral nucleus (Vo.r) using intra-axonal labeling with horseradish peroxidase and morphometric analyses. All labeled boutons contained round synaptic vesicles. Eighty-two percent of the boutons in the Vo.r and 99% of the boutons in the Vpd were presynaptic to nonprimary dendrites. The remaining boutons in the Vo.r were presynaptic to somata (8%) or primary dendrites (10%). The average number of postsynaptic profiles per labeled bouton did not differ in the Vpd and Vo.r. Most labeled boutons in the two nuclei were postsynaptic to unlabeled axon terminals with pleomorphic vesicles (p-ending). The number of p endings per labeled bouton was higher in the Vpd than Vo.r A morphometric analysis indicated that labeled bouton volume and apposed surface area were larger in the Vpd than Vo.r while active zone area and vesicle number did not differ. All these parameters were larger than those of p-endings in each nucleus. In both labeled boutons and p-endings, the parameters were positively correlated with bouton size. These results suggest that sensory information conveyed through trigeminal afferents is more strongly controlled at the level of the first synapse by presynaptic mechanisms in the Vpd than in the Vo.r, while the efficacy of transmission at primary afferent synapses does not differ. PMID- 11400177 TI - Reciprocal synaptic relationships between angiotensin II-containing neurons and enkephalinergic neurons in the rat area postrema. AB - A preembedding double immunostaining technique was used to study synaptic relationships between angiotensin-II-like immunoreactive and enkephalin-like immunoreactive neurons in the rat area postrema. The angiotensin-II-like immunoreactive neurons were detected by silver-gold intensification of the DAB reaction results while the enkephalin-like immunoreactive neurons were detected by simple ABC-DAB reaction. The synaptic relationships were reciprocal between the two neurons. Most of the synapses found between these two neurons were the presynaptic enkephalin-like immunoreactive axon terminals that made synapses on the angiotensin-II-like immunoreactive perikarya and dendrites. Both the axo somatic and axo-dendritic synapses were symmetrical. However, although angiotensin-II-like immunoreactive axon terminals also made synapses on enkephalin-like perikarya and dendrites, the axo-somatic synapses were symmetrical, while the axo-dendritic synapses were asymmetrical. The present results confirm the presence of angiotensin-II-like immunoreactive neurons in the area postrema and suggest that these angiotensinergic neurons in the area postrema may play a role in the regulation of blood pressure via coordinated synaptic interactions with enkephalinergic neurons. PMID- 11400178 TI - Cholinergic synaptic potentials in the supragranular layers of auditory cortex. AB - Receptive-field plasticity within the auditory neocortex is associated with learning, memory, and acetylcholine (ACh). However, the interplay of elements involved in changing receptive-fields remains unclear. Herein, we describe a depolarizing and a hyperpolarizing potential elicited by repetitive stimulation (20-100 Hz, 0.5-2 sec) and dependent on ACh, which may be involved in modifying receptive-fields. These potentials were recorded, using whole cell techniques, in layer II/III pyramidal cells in the rat auditory cortex in vitro. Stimulation at low stimulus intensities can give rise to a hyperpolarizing response and stimulation at higher stimulus intensities can elicit a depolarizing response. The depolarizing response had a reversal potential of -35 mV, and was reduced by the combination of AMPA/kainate and NMDA glutamate receptor antagonists (AMPA/kainate: CNQX, DNQX, and GYKI 52466; NMDA: APV, MK-801) and by the muscarinic ACh receptor antagonist atropine. The hyperpolarizing response had a reversal potential of -73 mV and could be reduced by atropine, GABA(A) receptor antagonists (bicuculline and a Cl(-) channel blocker picrotoxin), and to a small extent a GABA(B) receptor antagonist (saclofen). This suggests that the hyperpolarizing response is likely to be mediated by ACh acting on GABAergic interneurons. Extracellular recordings, also made from layer II/III of cortical slices, yielded a negative-going potential which was reduced by ionotropic glutamate receptor antagonists (same as above) and by the ACh receptor antagonists atropine and scopolamine, suggesting that this potential was the extracellular representation of the depolarizing response. PMID- 11400179 TI - Neuroprotective properties of 17beta-estradiol, progesterone, and raloxifene in MPTP C57Bl/6 mice. AB - Previous work from our laboratory showed prevention of 1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine (MPTP) induced dopamine depletion in striatum of C57Bl/6 mice by 17beta-estradiol, progesterone, and raloxifene, whereas 17alpha-estradiol had no effect. The present study investigated the mechanism by which these compounds exert their neuroprotective activity. The hormonal effect on the dopamine transporter (DAT) was examined to probe the integrity of dopamine neurons and glutamate receptors in order to find a possible excitotoxic mechanism. Drugs were injected daily for 5 days before MPTP (four injections, 15 mg/kg ip at 2-h intervals) and drug treatment continued for 5 more days. MPTP induced a decrease of striatal DAT-specific binding (50% of control) and DAT mRNA in the substantia nigra (20% of control), suggesting that loss of neuronal nerve terminals was more extensive than cell bodies. This MPTP-induced decrease of striatal [(125)I]RTI 121 specific binding was prevented by 17beta-estradiol (2 microg/day), progesterone (2 microg/day), or raloxifene (5 mg/kg/day) but not by 17alpha estradiol (2 microg/day) or raloxifene (1 mg/kg/day). No treatment completely reversed the decreased levels of DAT mRNA in the substantia nigra. Striatal [(125)I]RTI-121 specific binding was positively correlated with dopamine concentrations in intact, saline, or hormone-treated MPTP mice. Striatal NMDA sensitive [(3)H]glutamate or [(3)H]AMPA specific binding remained unchanged in intact, saline, or hormone-treated MPTP mice, suggesting the unlikely implication of changes of glutamate receptors in an excitotoxic mechanism. These results show a stereospecific neuroprotection by 17beta-estradiol of MPTP neurotoxicity, which is also observed with progesterone or raloxifene treatment. The present paradigm modeled early DA nerve cell damage and was responsive to hormones. PMID- 11400180 TI - Afferent synaptic contacts on glycine-immunoreactive neurons in the rat cuneate nucleus. AB - This study was aimed to clarify whether the primary afferent terminals (PATs), GABAergic terminals, and glutamatergic terminals made direct synaptic contacts with glycine-IR neurons in the cuneate nucleus of rats. In this connection, injection of the anterograde tracer WGA-HRP into brachial plexus, antiglycine preembedding immunoperoxidase, and anti-GABA, along with antiglutamate postembedding immunogold labeling, were used to identify the PATs, glycine-IR neurons, GABA-IR terminals, and glutamate-IR terminals, respectively. The present results showed that HRP-labeled PATs, immunoperoxidase-labeled glycine-IR terminals, immunogold-labeled GABA-IR, and glutamate-IR terminals made axodendritic synaptic contacts with immunoperoxidase-labeled glycine-IR neurons. The latter three presynaptic elements also formed axosomatic synapses with glycine-IR neurons. Statistical analysis has shown that the minimum diameter of the glycine-IR dendrites postsynaptic to the above-mentioned four presynaptic elements did not differ significantly. In addition, the synaptic ratio of the glutamate-IR terminals on the glycine-IR dendrites was higher than that of GABA IR terminals. The synaptic ratio of the GABA-IR terminals on glycine-IR dendrite was in turn higher than that of the PATs and glycine-IR terminals. It is suggested that the PATs and glutamate-IR terminals on the glycine-IR neurons may be involved in subsequent postsynaptic inhibition for spatial precision of lateral inhibition. On the other hand, the GABA-IR and glycine-IR terminals which make synaptic contacts with the dendrites of glycine-IR neurons may provide a putative means for disinhibition or facilitation to maintain the baseline neuronal activity in the rat cuneate nucleus. The results of quantitative analysis suggest that glutamate act as the primary excitatory neurotransmitter, while GABA, when compared with glycine, may serve as a more powerful inhibitory neurotransmitter on glycine-IR neurons in the rat cuneate nucleus. PMID- 11400181 TI - Effect of 5-HT on binding of [(11)C] WAY 100635 to 5-HT(IA) receptors in rat brain, assessed using in vivo microdialysis nd PET after fenfluramine. AB - By using a combination of positron emission tomography (PET) and postmortem tissue dissection, the effect of increased endogenous serotonin on specific binding of [(11)C]WAY 100635 to the 5-HT(1A) receptor was investigated in rat brain in vivo. The binding studies were complemented by in vivo microdialysis to monitor 5-HT levels in similarly treated isoflurane-anaesthetised rats, with the dialysis probe locations corresponding to two of the tissues sampled for specific binding of the radioligand. Fenfluramine treatment (10 mg/kg i.p.) resulted in a approximately 5-fold increase in extracellular 5-HT in medial prefrontal cortex and a approximately 15-fold increase in lateral hippocampus, maximal at approximately 40 min after injection. PET scan duration was either 60 or 90 min, beginning 30 min after fenfluramine injection. The specific binding of [(11)C]WAY 100635 was reduced by 10-20% in hippocampus, which showed highest binding in control animals. Specific binding, however, was unaffected in both prefrontal cortex and midbrain raphe, each additional high binding regions. The minimal effects are consistent with a low baseline occupancy of the 5-HT(1A) receptor by 5-HT in vivo, so that only a large change in endogenous agonist concentration will affect radioligand binding. This implies that utilisation of [(11)C]WAY 100635 in human PET to quantify 5-HT(1A) receptor expression can be extended to pathology where synaptic 5-HT levels are altered as a consequence of the disease state. PMID- 11400182 TI - SCH 58261, an A(2A) adenosine receptor antagonist, counteracts parkinsonian-like muscle rigidity in rats. AB - The aim of the present study was to find out whether blockade of adenosine A(2A) receptors by a selective antagonist, SCH 58261, influenced parkinsonian-like muscle rigidity. Muscle tone was examined using a combined mechano- and electromyographic method which simultaneously measured muscle resistance (MMG) of a rat hindfoot to passive extension and flexion in the ankle joint and electromyographic activity (EMG) of the antagonistic muscles of that joint: gastrocnemius and tibialis anterior. Muscle rigidity produced by reserpine (5 mg/kg + alpha-methyl-p-tyrosine, 250 mg/kg) was antagonized by SCH 58261 (0.1-5 mg/kg). SCH 58261 (5 mg/kg) also reduced reserpine-enhanced tonic and reflex EMG activities in both the gastrocnemius and the tibialis muscles. Moreover, SCH 58261 in doses of 1 and 5 mg/kg abolished muscle resistance induced by haloperidol (0.5 mg/kg). However, only the highest dose of SCH 58261 (5 mg/kg) decreased tonic EMG activity enhanced by haloperidol. Administration of L-DOPA (75 and 100 mg/kg) dose-dependently decreased the muscle resistance as well as tonic EMG activity evoked by haloperidol. Combined administration of SCH 58261 (0.1 mg/kg) and L-DOPA (50 mg/kg) in doses which did not affect the haloperidol induced muscle rigidity produced a pronounced synergistic effect. The ability of SCH 58261 to diminish the parkinsonian-like muscle rigidity and to potentiate the effect of L-DOPA in this model seems to indicate a therapeutic value of this compound in the treatment of Parkinson's disease. PMID- 11400183 TI - Acute neuroleptic stimulates DOPA decarboxylase in porcine brain in vivo. AB - The activity of DOPA decarboxylase measured in homogenates from rat striatum, or calculated from the rate of tracer decarboxylation measured ex vivo, is stimulated following acute treatment with antagonists of dopamine D2-like receptors. We used compartmental kinetics to test the hypothesis that utilization of the DOPA decarboxylase substrate [(18)F]fluorodopa is potentiated in living striatum following acute treatment with a typical neuroleptic. The kinetics of the tracer uptake were determined in eight anesthetized female pigs (40 kg) and in three animals receiving an infusion of haloperidol (75 microg kg(-1) h(-1)) for 1 h prior to tracer administration and throughout the 2-h positron emission recording. The relative activity of DOPA decarboxylase in striatum was increased threefold by the treatment. This potentiation of DOPA decarboxylation after pharmacological blockade of dopamine D2-like receptors may be used to optimize the utilization of exogenous DOPA in the treatment of Parkinson's disease. PMID- 11400184 TI - Anticocaine catalytic antibodies have no affinity for RTI compounds: implications for treatment. AB - Potential medications for cocaine abusers include: anticocaine catalytic antibodies, which could serve as circulating peripheral blockers of cocaine that prevents its action in the brain; and 3-phenyltropane cocaine analogs, which could serve as potent, selective, and long-lasting substitutes that reduce drug seeking. In order to evaluate the compatibility of these agents, we measured if a catalytic antibody would bind and interact with some cocaine analogs. Anticocaine catalytic antibody 15A10 had no significant affinity for RTI-51, RTI-112, or RTI 177 as examined by ELISA. They exhibited high affinity for the immunogen TSA1 in the same experiment, as expected. Because the antibody and the RTI compounds do not interact, they are candidates for simultaneous use. PMID- 11400185 TI - Absolute stereochemistry of guaianolides, of matricin and its epimers, of yarrow proazulenes, and of chamazulene carboxylic acid. AB - Known determinations of the absolute configuration of guaianolides were collected and found to be few. The absolute configurations of guaianolides rest on the assumption that 7-H always has alpha-orientation. For matricin, only the relative configuration was determined. On the basis of a detailed study of the NMR spectra of matricin and its epimers, and of synthetic, NMR, and CD studies with its decomposition product, chamazulene carboxylic acid, we were able to reconfirm the accepted 3S,3aR,4S,9R,9aS,9bS configuration of matricin. PMID- 11400186 TI - Stereoselective effects of (R)- and (S)-carvedilol in humans. AB - Carvedilol is currently used as the racemic mixture, (R,S)-carvedilol, consisting of equal amounts of (R)-carvedilol, an alpha-blocker, and (S)-carvedilol, an alpha- and beta-blocker, which have never been tested in their optically pure forms in human subjects. We performed a randomized, double-blind, placebo controlled, crossover study in 12 healthy male volunteers. Subjects received single oral doses of 25 mg (R,S)-carvedilol, 12.5 mg (R)-carvedilol, 12.5 mg (S) carvedilol, and placebo at 8 AM as well as at 8 PM. Exercise was performed at 11 AM, and heart rate and blood pressure were measured at rest and after 10 min of exercise. Urine was collected between 10 AM and 6 PM, as well as between 10 PM and 6 AM, and the amounts of urinary 6-hydroxy-melatonin sulfate (aMT6s) were determined by RIA. Compared to placebo, (R)-carvedilol increased heart rate during exercise (+4%, P < 0.05) and recovery (+10%, P < 0.05); (S)-carvedilol decreased heart rate during exercise (-14%, P < 0.05) and recovery (-6%, P < 0.05), and systolic blood pressure during exercise (-12%, P < 0.05); (R,S) carvedilol decreased heart rate during exercise (-11%, P < 0.05), and systolic blood pressure at rest (-7%, P < 0.05) and during exercise (-10%, P < 0.05). None of the agents had any significant effect on the release of aMT6s. Our results indicate that only (S)-carvedilol causes beta-blockade, whereas (R)-carvedilol appears to increase sympathetic tone, presumably as a physiological reaction to the decrease of blood pressure caused by alpha-blockade. None of the drugs had any influence on melatonin release. The weak clinical net effect of beta-blockade of (R,S)-carvedilol at rest might be one reason why this drug causes fewer side effects than other beta-blockers, such as a reduction of nocturnal melatonin release. PMID- 11400187 TI - Induced circular dichroism in solid-state inclusion compounds. AB - In order to measure the circular dichroism (CD) spectrum in the solid state of a chiral compound which has no chromophore, an induced CD spectral method was developed by measuring the spectrum of the inclusion crystal of the chiral compound with a simple achiral aromatic host compound in the Nujol mulls. PMID- 11400188 TI - Chiral asymmetry in spiral galaxies? AB - Spiral galaxies are chiral entities when coupled with the direction of their recession velocity. As viewed from the Earth, the S-shaped and Z-shaped spiral galaxies are two chiral forms. What is the nature of chiral symmetry in spiral galaxies? In the Carnegie Atlas of Galaxies that lists photographs of a total of 1,168 galaxies, we found 540 galaxies, classified as normal or barred spirals, that are clearly identifiable as S- or Z- type. The recession velocities for 538 of these galaxies could be obtained from this atlas and other sources. A statistical analysis of this sample reveals no overall asymmetry but there is a significant asymmetry in certain subclasses: dominance of S-type galaxies in the Sb class of normal spiral galaxies and a dominance of Z-type in the SBb class of barred spiral galaxies. Both S- and Z-type galaxies seem to have similar velocity distribution, indicating no spatial segregation of the two chiral forms. PMID- 11400189 TI - Chirality transferring [3,3] sigmatropic rearrangement of (1-Acyloxy-2 alkenyl)trianlkylsilane synthesis of optically active vinylsilane-containing alpha-amino acid. AB - A vinylsilane-containing alpha-amino acid and alpha,alpha-disubstituted alpha amino acid 2 having two contiguous asymmetric carbon centers at their alpha and beta positions were synthesized in an optically active form by ester-enolate Claisen rearrangement of the alpha-acyloxysilane 1 as the key step, where the chirality of an alpha-acyloxy-TBDMS group was completely transferred to the rearranged product. PMID- 11400190 TI - Asymmetric catalytic ene-cyclization approach to 2-fluoro-19-nor-1,25 dihydroxyvitamin D(3) A-ring analog with significant transactivation activity. AB - 1alpha,25-Dihydroxyvitamin D(3) (1alpha,25(OH)2D3) has been shown to modulate not only proliferation and differentiation, but also apoptosis in malignant cells, indicating that it could be useful for the treatment of cancer and psoriasis. However, little information has been available on the binding conformation of the 1alpha,25(OH)2D3 molecule and its analogs with the vitamin D receptor (VDR). Therefore, we synthesized 2alpha-fluorinated A-ring analogs of 19-nor 1alpha,25(OH)2D3 in order to investigate the VDR-binding conformation of the A rings on the basis of the (19)F NMR analysis. The 2alpha-fluoro-19-nor-1alpha,25 dihydroxyvitamin D3 A-ring analog thus synthesized via a asymmetric catalytic carbonyl-ene cyclization, shows significant activity in transactivation. PMID- 11400191 TI - Enantioselective inhibition of the binding of rac-profens to human serum albumin induced by lithocholate. AB - The reversible binding of lithocholate to human serum albumin determines a decrease of the binding of rac-ketoprofen. The process was followed by displacement chromatography using increasing concentrations of the competitor, i.e., lithocholate, in the mobile phase. The inhibition of rac-ketoprofen binding resulting was enantioselective and greater displacement was observed for the (S) enantiomer. The displacement process resulting was competitive in nature, the two enantiomers of ketoprofen binding to the same binding site as the modifier. The investigation was extended to other nonsteroidal antiinflammatory drugs. The enantioselective binding inhibition was larger in the case of rac-naproxen and rac-suprofen with respect to the phenomenon observed in the case of rac ketoprofen. The difference in circular dichroism spectroscopy was also used to characterize the binding of lithocholate to human serum albumin. This bile acid was proven to bind to site II on human serum albumin. The results, as obtained by displacement chromatography and difference circular dichroism spectroscopy, strongly support the hypothesized role of bile acids in inducing the enantioselective inhibition of ketoprofen binding to human serum albumin in patients suffering from liver diseases. PMID- 11400192 TI - Novel asymmetric oxy-michael addition reaction of the chiral ketones to the achiral gamma--or delta-hydroxy-alpha,beta-unsaturated carbonyl compounds. AB - A novel asymmetric oxy-Michael addition reaction was developed. In the presence of a catalytic amount of base, chiral ketones 1 and 2, derived from D-glucose and D-fructose, respectively, reacted with omega-hydroxy enones or enoates 3a-e, 17 and 21 to form the hemiacetal-derived alkoxide which underwent stereoselective intramolecular Michael addition to give cyclic acetals. Although the stereoselectivities in the formation of the five-membered acetal rings were modest, six-membered ring formation proceeded with high stereoselectivity and the utility of the reaction was demonstrated by a simple syntheses of natural products. PMID- 11400194 TI - High-throughput semi-automated 96-well liquid/liquid extraction and liquid chromatography/mass spectrometric analysis of everolimus (RAD 001) and cyclosporin a (CsA) in whole blood. AB - A semi-automated high-throughput liquid/liquid extraction (LLE) assay was developed for RAD001 and cyclosporin A (CsA) in human blood. After addition of internal standard and ammonium hydroxide, samples were extracted twice with methyl tert-butyl ether (MTBE). The organic extract was evaporated to dryness and reconstituted in mobile phase. Where possible, sample transfer and LLE steps were automated using a Tomtec Quadra 96 workstation. Samples were analyzed using ESI LC/MS/MS employing the transitions of ([M + NH(4)](+) --> [M + H](+)) for CsA and ([M + NH(4)](+) --> [M + H-(CH(3)OH + H(2)O)](+)) for RAD001, under isocratic chromatographic conditions (75:25, (v/v), acetonitrile/20 mM ammonium acetate) with a run time of 3.6 min. A lower limit of quantitation (LLOQ) of 0.368 ng/mL and 5.23 ng/mL was achieved for RAD001 and CsA, respectively, using a sample volume of 0.3 mL for the analysis. The method was validated over a 3-day period and the resulting calibration curves had a correlation coefficient >0.99 over the concentration range 0.368 to 409 ng/mL and 5.24 to 1748 ng/mL for RAD001 and CsA, respectively. The inter-day coefficient of variation (CV) was less than 15% at the LLOQ for both compounds. The method was applied to the analysis of clinical samples. Under normal working conditions four 96-well plates could be extracted and LC/MS analysis completed in less than 28 h. A marked improvement in sample throughput efficiency was realized with this LLE method when compared to existing solid phase extraction (SPE) methods which deal with both RAD001 and CsA. PMID- 11400195 TI - Electrospray tandem mass spectrometry of lexitropsins. AB - Several compounds, representative of the class of lexitropsins, were analyzed by electrospray tandem mass spectrometry. The study of the fragmentations of the protonated molecular species ([M + H](+)) and of selected fragment ions allowed proposals for the main fragmentation pathways of compounds of this type. The interpretation of the fragmentation pathways of these compounds was complicated because of intramolecular hydrogen migration. In order to better understand the fragmentation pathways, the MS/MS/MS spectra of several compounds, and the MS/MS and MS/MS/MS spectra of the deuterated compounds, were obtained. Accurate mass measurements helped elucidate the structures of smaller fragment ions. Low-energy collision-induced decomposition (CID) tandem mass spectrometry of lexitropsins with electrospray ionization has proven to be a good method for the structural characterization and identification of this class of compounds. Main fragmentation pathways occur by cleavage of the peptide bond followed by the elimination of the substituted pyrrole ring, and their elucidation will facilitate structural characterization of new lexitropsins. PMID- 11400196 TI - Determination of gatifloxacin in human plasma by liquid chromatography/electrospray tandem mass spectrometry. AB - Gatifloxacin is an advanced-generation, 8-methoxyfluoroquinolone that is active against a broad spectrum of pathogens, including antiobiotic resistant Streptococcus pneumoniae. Development of a rapid, sensitive and selective method for the determination of gatifloxacin in human plasma is essential for understanding the pharmacokinetics of the drug when administered orally or intravenously. Solid phase extraction (SPE) using Oasis HLB was used to extract gatifloxacin and the internal standard ciprofloxacin from plasma. A method based on liquid chromatography/electrospray tandem mass spectrometry (LC/ESI-MS/MS) was developed and validated to quantitate gatifloxacin in human plasma. The precursor and major product ions of the analyte were monitored on a triple quadrupole mass spectrometer with positive ion electrospray ionization (ESI) in the multiple reaction monitoring (MRM) mode. Mechanisms for the formation of collision-induced dissociation products of gatifloxacin are proposed. Linear calibration curves were generated from 10--1000 ng/mL with coefficients of determination greater than 0.99. The interday and intraday precision (%RSD) was less than 6.0% and accuracy (%error) was less than 5.4% for gatifloxacin. The limit of detection (LOD) for the method was 500 pg/mL based on a signal-to-noise ratio of 3. PMID- 11400197 TI - Aldehyde analysis by high performance liquid chromatography/tandem mass spectrometry. AB - This paper describes the development of a high performance liquid chromatography/tandem mass spectrometric (MS/MS) procedure for the specific qualitative and quantitative analysis of lipid aldehydes in biological matrices. A derivatisation method, which results in molecules that exhibit a common product ion on MS/MS, permits informative precursor ion scans, at high sensitivity. This has been applied to the examination of plasma in order to examine the production of aldehydes consequent on in vitro lipid oxidation. Quantitative analysis of target molecules using multiple reaction monitoring has been developed to permit quantitation in the same matrices. PMID- 11400198 TI - Liquid chromatography/atmospheric pressure chemical ionization mass spectrometry of terpene lactones in plasma of volunteers dosed with Ginkgo biloba L. extracts. AB - Liquid chromatography/atmospheric pressure chemical ionization mass spectrometry (LC/APCI-ITMS) was applied to evaluate the levels of ginkgolides A and B and bilobalide in plasma of volunteers after administration of Ginkgo biloba extracts in free (Ginkgoselect) or phospholipid complex (Ginkgoselect Phytosome) forms, providing 9.6 mg of total terpene lactones. The maximum plasma concentrations, C(max), of total ginkgolides A, B and bilobalide were 85.0 and 181.8 microg/mL for Ginkgoselect and Ginkgoselect Phytosome, respectively. The C(max) values were reached at 120 min for the free form and at 180--240 min for the phospholipid complex form. In both cases, the mean elimination half-life of each terpene lactone was in the range 120--180 min. Due to its sensitivity (about 1 ng/mL) and specificity, LC/APCI-ITMS proved to be a very powerful tool for pharmacokinetic studies of these phytochemicals. PMID- 11400199 TI - Analysis of plant phosphatidylcholines by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - The use of matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) for the quantitative determination of phospholipid (PL) molecular species has been problematic, due primarily to the formation of multiple signals (corresponding to the molecular ion and other adducts) for some classes of PL. For example, analysis of phosphatidylcholine (PC) yielded signals that corresponded to protonated and sodiated molecules in the MALDI spectrum. The resulting spectral overlap among various molecular species (e.g. [PC(16:0/18:2) + Na] and [PC(18:2/18:3)]) made it impossible to ascertain their relative amounts using this technique. Other spectral ambiguities existed among different structural isomers, such as PC(18:1/18:1) and PC(18:0/18:2). We determined that molecular species could be resolved by MALDI-TOFMS by first removing the polar head (e.g. phosphocholine) from the phospholipid to effect production of only the sodiated molecules of the corresponding diacylglycerols (DAGs). Analysis of the resulting spectrum allowed unequivocal determination of the molecular species profile of PC from potato tuber and soybean. Estimation of fatty acid composition based on the molecular species determined by MALDI-TOFMS analysis agreed with that from GC-FID analysis. Post-source decay (PSD) was used to resolve standard isomers of PC (e.g. 18:1/18:1 vs. 18:0/18:2). Our results indicated that PSD is a useful approach for resolving structural isomers of PL molecular species. PMID- 11400203 TI - Estimation of proton affinity of proline and tryptophan under electrospray ionization conditions using the extended kinetic method. AB - The relative order of the proton affinity (PA) of 20 naturally occurring amino acids has been determined under electrospray ionization conditions and compared with earlier studies of different research workers. The order we obtained is similar to that reported by other groups except in three cases viz., valine- aspartic acid, asparagine--glutamic acid and tryptophan--proline. The PA values of proline and tryptophan are determined by the extended kinetic method using amino acids themselves as reference bases. The PA values we thus obtained for proline and tryptophan are 219.9 and 221.6 kcal/mol, respectively. PMID- 11400204 TI - Expanding the linear dynamic range in quantitative high performance liquid chromatography/tandem mass spectrometry by the use of multiple product ions. AB - A strategy for expanding the linear working range in bioanalysis using quantitative high performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS) is presented. The strategy involves monitoring multiple product ions. Herein we demonstrate the strategy on a rat plasma assay for a proprietary experimental drug where the linear range is expanded from 2 to 4 orders of magnitude. A primary sensitive ion was monitored to obtain a high sensitivity range calibration curve (0.400 to 100 ng/mL) while a less sensitive secondary ion was monitored to obtain a low sensitivity range calibration curve (90.0 to 4000 ng/mL). Each calibration curve gave acceptable linearity (r >0.990). Quality control samples at low, mid and high levels within each calibration curve demonstrated acceptable precision and accuracy (within 20% for all levels). The technique was successfully applied to rat pre-clinical sample analysis. PMID- 11400205 TI - Comparison of electron capture dissociation and collisionally activated dissociation of polycations of peptide nucleic acids. AB - Electron capture dissociation (ECD) in Fourier transform ion cyclotron resonance mass spectrometry coupled with electrospray ionization enhances the sequence elucidation of peptide nucleic acids compared with conventional low-energy collisionally activated dissociation (CAD). Examples are shown where ECD produced complete or extensive sequence coverage in PNAs six to ten nucleobases long. However, facile base losses from the reduced species and low abundances of backbone ECD fragments presented a significant problem. This was rationalized through the lower degree of charge solvation on the backbone compared to polypeptides. Combination of both CAD and ECD data is advantageous, as these techniques produce cleavages at different sites. PMID- 11400206 TI - Preliminary study of the analysis of oligogalacturonic acids by electrospray ionization mass spectrometry. AB - During systematic studies of the behavior of oligogalacturonic acids (OGAs) under different conditions using electrospray ionization mass spectrometry (ESI-MS), cation adduction, fragmentation and non-covalent binding were found to be the three major problems that compromised the analysis of OGAs by ESI-MS. By adjusting solution components, capillary temperature and capillary voltage in the ESI source, an optimized condition was found that gave a clean and clear spectrum of trigalacturonic acid. A direct injection ESI-MS technique based on the use of aqueous acetonitrile and acetic acid and triethylamine (TEA) as modifiers has been applied to analyze a mixture including mono-, di- and trigalacturonic acids, which will facilitate further applications of ESI-MS in the analysis of mixtures of OGAs. PMID- 11400208 TI - Direct analysis of pharmaceutical compounds in human plasma with chromatographic resolution using an alkyl-bonded silica rod column. AB - Monolithic columns have been successfully used with steep gradient and high flow rates for the direct analysis of a candidate pharmaceutical compound in human plasma. The monolithic columns showed excellent robustness with nearly 300 20 microL injections of plasma (diluted 1:1 with water) being made onto one column without significant deterioration in performance. The system gave excellent sensitivity with a limit of quantification of 5 ng/mL being achieved. Unlike previous methods of direct analysis the monolithic columns showed excellent resolution even after nearly 300 plasma injections. The column performance was measured before and after the analysis of the plasma samples. PMID- 11400209 TI - Increased throughput in quantitative bioanalysis using parallel-column liquid chromatography with mass spectrometric detection. AB - The feasibility of quantitative bioanalysis by parallel-column liquid chromatography in conjunction with a conventional single-source electrospray mass spectrometer has been investigated using plasma samples containing a drug and its three metabolites. Within a single chromatographic run time, sample injections were made alternately onto each of two analytical columns in parallel at specified intervals, with a mass spectrometer data file opened at every injection. Thus, the mass spectrometer collected data from two sample injections into separate data files within a single chromatographic run time. Therefore, without sacrificing the chromatographic separation or the selected reaction monitoring (SRM) dwell time, the sample throughput was increased by a factor of two. Comparing the method validation results obtained using the two-column system with those obtained using the corresponding conventional single-column approach, the methods on the two systems were found to be equivalent in terms of accuracy and precision. The parallel-column system is simple and can be implemented using existing laboratory equipment with no additional capital outlays. A parallel column system configured in this manner can be used not only for the within-a-run analysis of two samples containing two different sets of chemical entities, but also for the within-a-run analysis of two samples containing the same set of chemical entities. PMID- 11400210 TI - Electrospray ionisation tandem mass spectrometry in the characterisation of isomeric benzofurocoumarins. AB - A set of aminoalkoxy-substituted, differently annullated furocoumarins, differing in the position of the aminoalkoxy chain and in the unsaturation level of the fused ring, has been subjected to electron impact and electrospray ionisation (ESI) experiments. In order to achieve a distinct characterisation of isomeric compounds, which partially failed under electron impact conditions, collision induced dissociation experiments were performed on protonated molecules. The breakdown curves obtained by varying the tickle voltage on an ion trap ESI instrument led to the desired characterisation. PMID- 11400211 TI - Modulation effects of zinc on the formation of vitamin D receptor and retinoid X receptor alpha-DNA transcription complexes: analysis by microelectrospray mass spectrometry. AB - The vitamin D receptor (VDR) binds zinc, and the activity of vitamin D dependent genes in cells is influenced by intracellular zinc concentrations. To determine whether zinc influences vitamin D action in cells by modulating the formation of VDR and retinoid x receptor alpha (RXR alpha) heterodimer-DNA complexes, we used microelectrospray ionization mass spectrometry (microESI-MS) to assess receptor DNA interactions in the presence of varying amounts of zinc. In the absence of DNA, VDR and RXR alpha proteins were primarily monomeric with small amounts of protein homodimers also observed. Zn(2+) (up to 300 microM) did not change VDR or RXR alpha monomer/homodimer ratios. Mass spectra of VDR combined with RXR alpha were a sum of individual protein spectral data. Zn(2+) had no effect on the interactions of receptors. With increasing amounts of Zn(2+), additional Zn(2+) ions were detected bound to VDR and RXR alpha. microESI-MS analyses of RXR alpha in the presence of an osteopontin vitamin D DNA response element (OP-VDRE) showed RXR alpha homodimer/OP-VDRE complexes. DNA-protein complex formation increased on addition of Zn(2+) up to 200 microM; at 300 microM, Zn(2+) dissociation of the RXR alpha homodimer/OP-VDRE complexes occurred, coincident with the appearance of RXR alpha monomeric protein. When microESI-MS analyses were carried out with VDR and OP-VDRE, VDR homodimer/OP-VDRE complexes were not detected. Addition of Zn(2+) did not result in VDR/OP-VDRE complex formation. Heterodimeric VDR/RXR alpha complexes with OP-VDRE were detected by microESI-MS. Addition of 300 microM Zn(2+) resulted in dissociation of the heterodimeric VDR/RXR alpha/OP-VDRE complex. Addition of Mg(2+) in place of Zn(2+) did not alter protein/OP-VDRE complexes. Our results show that zinc modulates steroid hormone receptor-DNA interactions. PMID- 11400212 TI - Electrospray ionisation tandem mass spectrometric study of hydrogen-bonding interactions of some disaccharides with lysine. AB - The efficiency of formation of protonated heterotrimers of lysine with underivatised sugars (mono-, di-, and trisaccharides) and N-acetylglucosamine (N AcGlc) was studied under electrospray ionisation conditions. The collision induced dissociation spectra of [Lys + sugar + NAcGlc + H](+) resulted in [Lys + NAcGlc + H](+) and [Lys + sugar + H](+) as the major product ions. Relative abundances of these two fragments reflect the extent of adduct formation of protonated lysine plus sugar, with reference to the reference compound NAcGlc. This relative abundance ratio was found to be characteristic of the sugar structure. In this way it was observed that the ability of lysine to form a protonated heterodimer with neutral sugars increases with an increase in the number of acetal oxygens. Lactose showed an anomalously high affinity for protonated Lys, possibly reflecting the axial hydroxyl at C4. The postulated involvement of the epsilon-NH(2) group of lysine in the formation of protonated heterodimers with sugars was supported by similar results of similar experiments with NH(3) in place of lysine. PMID- 11400213 TI - The early glycation products of the Maillard reaction: mass spectrometric characterization of novel imidazolidinones derived from an opioid pentapeptide and glucose. AB - Glucose-substituted imidazolidinones related to the endogenous opioid peptide leucine-enkephalin have been investigated using fast atom bombardment tandem mass spectrometry (FAB-MS/MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS). In addition to Amadori compounds, the studied imidazolidinones represent a novel type of the early glycation products formed in the Maillard reaction. To obtain insight into the fragmentation behavior of these carbohydrate peptide adducts, we also studied synthetic precursors of the glucose-substituted imidazolidinones as well as the corresponding isopropylidene derivatives. The collision-induced dissociation (CID) spectra of [M + H](+) ions of all these imidazolidinones have been compared. Detailed analysis showed that fragmentation of each compound generates two ions at m/z 566 and m/z 598 which are characteristic and undoubtedly confirm the imidazolidinone-type structure. These two significant ions were identified as the M + 10 and M + 42 modifications of the N-terminus of the parent opioid pentapeptide effected by the carbohydrate moiety. Furthermore, the ion at m/z 178 is identified as the M + 42 modification of the immonium ion of the N-terminal amino acid (tyrosine) also effected by the carbohydrate moiety. They can be used as diagnostic ions for imidazolidinone-type compounds in studying the Maillard reaction. Thus, we have demonstrated the utility of FAB-MS/MS and ESI-MS/MS in the structural determination and identification of such novel peptide-carbohydrate adducts, useful in understanding the details of the mechanism of non-enzymatic glycation in vivo. PMID- 11400214 TI - Menopause as a measure of population health: an overview. PMID- 11400215 TI - A critique of the grandmother hypotheses: old and new. AB - The singularity of reproductive senescence in human females has led many investigators to consider menopause an adaptation permitting increased maternal investment in existing progeny. Much of the focus has been on the grandmother hypothesis-the notion that aging women gain an inclusive fitness advantage from investing in their grandchildren. This hypothesis has evolved from an explanation for menopause into an explanation for the exceptionally long postreproductive lifespan in human females. In the old grandmother hypothesis, menopause is an adaptation facilitating grandmothering; it is about stopping early in order to create a postreproductive lifespan. In the new grandmother hypothesis, grandmothering is an adaptation facilitating increased longevity, and menopause is a byproduct. This paper reviews and critically evaluates the evidence for and against both hypotheses, focusing on key predictions of each. If menopause is the result of selection for increased maternal investment, this involved mainly mothers, not grandmothers. PMID- 11400216 TI - Physiology of hot flashes. AB - Hot flashes are the most common symptom of the climacteric, although prevalence estimates are lower in some rural and non-Western areas. The symptoms are characteristic of a heat-dissipation response and consist of sweating on the face, neck, and chest, as well as peripheral vasodilation. Although hot flashes clearly accompany the estrogen withdrawal at menopause, estrogen alone is not responsible since levels do not differ between symptomatic and asymptomatic women. Until recently it was thought that hot flashes were triggered by a sudden, downward resetting of the hypothalamic setpoint, since there was no evidence of increased core body temperature. Evidence obtained using a rapidly responding ingested telemetry pill indicates that the thermoneutral zone, within which sweating, peripheral vasodilation, and shivering do not occur, is virtually nonexistent in symptomatic women but normal (about 0.4 degrees C) in asymptomatic women. The results suggest that small temperature elevations preceding hot flashes acting within a reduced thermoneutral zone constitute the triggering mechanism. Central sympathetic activation is also elevated in symptomatic women which, in animal studies, reduces the thermoneutral zone. Clonidine reduces central sympathetic activation, widens the thermoneutral zone, and ameliorates hot flashes. Estrogen virtually eliminates hot flashes but its mechanism of action is not known. PMID- 11400217 TI - Menstrual cycle variability and the perimenopause. AB - Menopause, the final cessation of menstrual cycling, occurs when the pool of ovarian follicles is depleted. The one to five years just prior to the menopause are usually marked by increasing variability in menstrual cycle length, frequency of ovulation, and levels of reproductive hormones. Little is known about the mechanisms that account for these characteristics of ovarian cycles as the menopause approaches. Some evidence suggests that the dwindling pool of follicles itself is responsible for cycle characteristics during the perimenopausal transition. Another hypothesis is that the increased variability reflects "slippage" of the hypothalamus, which loses the ability to regulate menstrual cycles at older reproductive ages. This paper examines the underlying cause of the increasing variability in menstrual cycle length prior to the menopause. A model of ovarian cycles is developed, based on the process of follicular growth and depletion. Under this model, the follicular phase of each menstrual cycle is preceded by an inactive phase, a period of time when no ovarian follicles have left the resting state and begun secreting steroids in response to gonadotropin stimulation. The model makes predictions about the variability in menstrual cycles across the reproductive life span based on the size of the surviving pool of ovarian follicles. We show that the model can explain several characteristics of the perimenopause in humans and macaques and illustrate how the model can be applied to research on the biological and cultural correlates of the timing of menopause. PMID- 11400218 TI - Marital status and age at natural menopause: considering pheromonal influence. AB - Married women generally report a later mean age at menopause. The results reported here, from a study carried out in Greene County, New York, are no exception. Married and widowed women report a later mean age at natural menopause compared to single and divorced women (P < 0.05). To better understand the relationship between marital status and age at menopause, possible mechanistic and confounding variables are examined, in particular parity, sexual activity, smoking habits, level of education, and income. Parity and income 10 years prior to interview are significant factors, along with marital status, that explain part of the variation in age at natural menopause. An alternative explanation is the pheromonal influence of a male in the household. This would explain the consistency of results across populations. This pilot study supports further biochemical investigation. PMID- 11400219 TI - Effects of age, ethnicity and menopause on ambulatory blood pressure: Japanese American and Caucasian school teachers in Hawaii. AB - Ambulatory blood pressure (BP) measurements of 120 female teachers of Japanese American or Caucasian ethnicity working in public schools located in Hilo, Hawaii, were recorded. BP was measured at 15-min intervals during waking hours and 30-min intervals during sleep over a 24-hr period that included a full work day. These measurements were averaged during three daily settings: at work, at home while awake ("home"), and during sleep. ANCOVAs using ethnicity as a predictor variable of BP, with age and the body mass index (BMI) as covariates, show a significant interaction effect between age and ethnicity in some daily settings. Among Japanese-Americans partial correlations between age and systolic BP controlling for the BMI are significant in these settings, while among Caucasians none of the correlations are significant. Menopausal status is not significantly related to BP when age is controlled in analyses. There was no significant ethnic difference in number of symptoms reported, including frequency of "hot flushes/flashes," within the past two weeks. Those who reported hot flushes had significantly elevated BP in waking settings but not during sleep. The greater increase in BP with age in Japanese-Americans may be related to their elevated risk for development of hypertension. The lack of a significant relationship between menopausal status and BP may be due to the high rate of usage of hormonal replacement therapy in this sample, as well as an unusually high rate of hysterectomy. PMID- 11400220 TI - Menopause, local biologies, and cultures of aging. AB - Menopause marks the end of menstruation, once generally accepted as the closure of women's reproductive lives. The current medical view of menopause, however, is as a pathological event with its own distinct set of symptoms and diseases. Researchers have described women as facing a dramatic increase in the risk of heart disease, osteoporosis, stroke, and Alzheimer's, all as the result of the impact of changing hormone levels, particularly the decline in estrogen. The clinical literature has interpreted these findings in terms of the absolute necessity of replacing these lost hormones for all women who are menopausal regardless of any other physiological, social, or cultural characteristic they might possess. Using research done in Japan, Canada, and the United States, this paper challenges the notion of a universal menopause by showing that both the symptoms reported at menopause and the post-menopause disease profiles vary from one study population to the next. For most of the symptoms commonly associated with menopause in the medical literature, rates are much lower for Japanese women than for women in the United States and Canada, although they are comparable to rates reported from studies in Thailand and China. Mortality and morbidity data from these same societies are used to show that post-menopausal women are also not equally at risk for heart disease, breast cancer, or osteoporosis. Rather than universality, the paper suggests that it is important to think in terms of "local biologies", which reflect the very different social and physical conditions of women's lives from one society to another. PMID- 11400221 TI - Menopausal experiences and bone density of Mayan women in Yucatan, Mexico. AB - This cross-sectional study reports on the menopausal transition of Mayan women from Yucatan, Mexico. A total of 228 women completed the study, and 118 women were classified by history as postmenopausal; the others were premenopausal. Demographic information, reproductive history, physical examination, hormone concentrations, radial bone density, food samples, and history of physical activity were obtained. The average age at which menopause occurred by history was 44.3 +/- 4.4 years; this is reflected in the distribution of FSH levels by age. None of the women reported symptoms of hot flashes, and none recalled any history of significant symptoms associated with their menopausal transition. Hormone levels were similar to U.S. reference values with elevated FSH (66.6 +/- 29.1 mIU/ml), low estradiol (9.4 +/- 8.3 pg/ml) and estrone (13.3 +/- 7.8 pg/ml), E1 > E2, and normal levels of testosterone and androstenedione. BMD declined with age, and values were lower than reference values for United States women. Clinical evidence of fracture was not detected by history or physical examination even for those who were 20 years postmenopausal. The endocrine characteristics of menopause among Mayan women in Yucatan are similar to hormonal changes reported for women in the United States, but signs, symptoms, and apparent consequences are different in the two populations. PMID- 11400222 TI - Associations with age at natural menopause in Blackfeet women. AB - This study seeks to determine the age at natural menopause and factors that may predict that age for Blackfeet women living on the reservation in Montana. Data were obtained from a probability sample of 150 women. Rates of surgical menopause among women who had experienced menopause were similar to those found in NHANES I for U.S. women. Median age at natural menopause by the status quo method (51.2 years) was comparable to that determined for other populations. Within the group of women who already had experienced natural menopause, age at menopause was associated in bivariate models with age at menarche (b = -0.83 +/- 0.44 years, P = 0.07), household income (b = 1.85 +/- 0.71 years, P = 0.01), "ever use" of birth control (b = 4.52 +/- 1.58 years, P = 0.008), "ever use" of oral contraceptives (b = 5.48 +/- 1.91 years, P = 0.007), current or recent unemployment (b = 4.75 +/- 1.97 years, P = 0.02), having been breastfed (b = 6.93 +/- 2.67 years), and educational attainment (ANOVA by tertiles of menopause age, mean 10.0 +/- 3.6 years of education for earliest, 13.0 +/- 2.2 years for latest, P = 0.04). In multivariate models, "ever use" of oral contraceptives, household income, and age at menarche remained significant predictors of age at menopause (R2 = 0.33-0.35, P = 0.002-0.004), with household income modifying the effect of age at menarche (analysis ofjoint effects). The findings are discussed in the context of cohort experiences, examining implications for both early and later-life influences on menopause age. PMID- 11400223 TI - Postmenopausal function in context: biocultural observations on Amish, neighboring non-Amish, and Ifugao household health. AB - Ecological approaches to female post-reproductive function must consider the social context and microenvironment of the household. A model integrating these environmental factors into the analysis of late-life well-being (operationally defined as household health) is presented and explored in three settings: an Old Order Amish community, the neighboring community of non-Amish, and an Ifugao community in the Philippine highlands. Intensive interview and observation of function were completed in a sample of at least 25 households in each community. The complex interaction of social and biological factors is explored in case studies and through multiple regression analysis with household health as the dependent variable. The results emphasize the importance of social support and assessing capabilities within the household. The results also suggest the need for natural field observation of work to better understand the influence of pacing and scheduling, necessity of work, and the role of environmental modification in postmenopausal function. PMID- 11400224 TI - Anger expression, body fat, and blood pressure in adolescents: Project HeartBeat! AB - This study explores the potential influence of growth, body/composition, and sexual maturity on the relation of anger expression and blood pressure in adolescents. Baseline data from Project HeartBeat! (82 boys and 85 girls, 14 years of age) examined the ability of anger expression (STAXI scale) to predict blood pressure, after controlling for the effects of ethnicity (African-American/ non-African-American), height, weight, percentage body fat, and sexual maturity. Blood pressures were unrelated to anger expression in models that included the above developmental variables. However, girls scoring high on healthy anger expression ("anger-control") had significantly lower levels of percentage body fat (P = 0.015) independent of the above factors. The literature suggests that body fat or body mass is often, though not unanimously, associated with unhealthy forms of anger expression in adolescents. Research is required into the biological, social, and behavioral origins of the association between body fat and anger expression. Height and sexual maturity, virtually ignored in this literature, should be included in future research. PMID- 11400225 TI - Body size, composition, and blood pressure of high-altitude Quechua from the Peruvian Central Andes (Huancavelica, 3,680 m). AB - Although much information is available about the effects of high altitude on physiological characteristics, less is know about its effect on body composition. In the present study, anthropometric and body composition variables were investigated in a sample of 77 adult Quechua males from the Peruvian Central Andes (Huancavelica, 3,680 m). The subjects are shorter in relation to body weight than other ethnic groups, whereas body proportions are macrocormic (indicating a long trunk relative to the lower extremities), with intermediate values of the acromial-iliac index. All skinfold thicknesses are low (approximately 15th percentiles of NHANES reference values for the triceps and subscapular skinfolds), but tend to be higher than in the other Quechua populations. Similar results are obtained when percentage fat is estimated. Somatotypes are dominant in mesomorphy with very low ectomorphy. Comparison with a sample of high-altitude Kirghiz (3,200 m), previously studied with the same methods, shows higher values in the Peruvian sample for all variables related to adiposity. The presence of low adiposity in the Quechua population could be associated with stresses of the high-altitude environment. Mean values of blood pressure are very low and there is no correlation with age. PMID- 11400226 TI - Child factor in measurement dependability. AB - A primary consideration in longitudinal growth studies is the identification of growth from error components. While previous research has considered matters of measurement accuracy and reproducibility in detail, few reports have investigated the errors of measurement due to aspects of the physiology and cooperation of the child. The present study directly assesses this source of measurement undependability for the first time. Investigation of total measurement error variance in 925 recumbent length replicates taken over stasis intervals in growth identifies that between 60% and 70% of total measurement unreliability is due to a child factor undependability. Individual differences are significant and longitudinal growth analyses should consider two to three times the technical error of measurement statistic as a reasonable estimate of the total unreliability for any single measurement of an infant's recumbent length. These results raise issues regarding analytic methods as applied to serial growth data. PMID- 11400227 TI - Age and survival from squamous cell carcinoma of the oral tongue. AB - BACKGROUND: A worse outcome for young patients with head and neck squamous cell carcinoma has been previously suggested in the literature. This issue has been investigated with respect to squamous cell carcinoma of the oral tongue. METHODS: The Surveillance, Epidemiology and End Results (SEER) program tumor registries were used. Cases of squamous cell carcinoma of the oral tongue (ICD-9 codes 141.1 141.5) diagnosed from 1988-1993 in which this cancer was the one and only cancer were included (n = 749). Disease-specific survival was evaluated with respect to age, type of surgical treatment, and radiotherapy while stratifying for stage using Cox proportional hazards analysis. A secondary analysis included the additional variables, tumor size and nodal status. (These fields were recorded in SEER for only about half of the cases in the primary analysis.) RESULTS: Analysis revealed that increasing age predicted worse disease-specific survival. A 10-year increase in age was associated with an 18% increase in risk of death. Surgical therapy with excision of the primary tumor alone or excision plus neck dissection predicted improved survival, whereas the use of radiotherapy was associated with worse survival. (The latter may reflect bias associated with positive surgical margins or premorbid conditions.) In the secondary analysis, age, tumor size, and positive lymph node status were associated with significantly worse disease specific survival, whereas surgical excision plus neck dissection was associated with improved survival. CONCLUSION: The SEER database shows increased disease specific mortality with increasing age in patients with cancer of the oral tongue. Surgical therapy is associated with improved survival, whereas the use of radiotherapy, increasing tumor size, and positive lymph node status are associated with worse outcome. PMID- 11400228 TI - Prognostic significance of Bcl-2 and p53 expression in advanced laryngeal squamous cell carcinoma. AB - BACKGROUND: Proteins regulating the cell cycle and cell death are frequently abnormally expressed in cancer. Several of these, particularly p53 and Bcl-2, have been widely suggested as possible prognostic markers in diverse human malignancies. Their role in predicting outcome in squamous cell carcinomas of the head and neck is unclear and may depend on the location, stage, and treatment of the tumor. METHODS: To assess this question specifically for advanced squamous cell carcinoma of the larynx, we studied 69 patients with stage III or IV tumors, all but 6 of whom were treated with surgery plus postoperative irradiation by a single physician. We studied the patients retrospectively to test the association between expression of Bcl-2 and p53, as assessed by immunohistochemistry, with treatment outcome and survival. RESULTS: Twenty of the 69 patients died from their tumor (poor outcome); the rest were alive and tumor free at the last follow up or died of unrelated causes without clinical tumor recurrence (good outcome). Fourteen tumors had detectable Bcl-2 expression, including 8 scored as overexpressors. Thirty-nine tumors overexpressed p53. Expression of neither Bcl-2 nor p53 was associated with outcome, overall survival, or disease-free survival. Only tumor stage was significantly associated with outcome and disease-free survival. CONCLUSION: These data indicate that assessing expression of p53 or Bcl 2 is unlikely to be prognostically useful for surgically treated advanced laryngeal carcinoma. PMID- 11400229 TI - Prognostic significance of serum p53 protein and p53 antibody in patients with surgical treatment for head and neck squamous cell carcinoma. AB - OBJECTIVES: This study aims at investigating the prognostic values of serum p53 protein and anti-p53 antibody in patients undergoing surgical treatment for head and neck squamous cell carcinoma (HNSCC). METHODS: Serum p53 protein and anti-p53 antibody concentrations were determined by an enzyme-linked immunosorbent assay (ELISA) in 75 patients with HNSCC and 28 healthy controls. In 28 patients, formalin-fixed tumor tissues were also available for immunohistochemical staining by an anti-p53 DO7 monoclonal antibody. The results were correlated with the clinicopathologic parameters. RESULTS: The mean preoperative serum concentration of p53 protein in patients with HNSCC was significantly higher than healthy controls (59.45 pg/mL vs 16.4 pg/mL, p =.007). Preoperative serum p53 antibody was present in 23 (31%) patients and was present in one healthy control. Eighteen (62%) tumor tissues showed p53 overexpression by immunohistochemistry. The presence of serum anti-p53 antibody before operation was associated with a significantly higher incidence (65%) of nodal metastasis compared with 27% nodal metastasis in patients with absence of serum anti-p53 antibody (p =.002). CONCLUSION: Preoperative serum p53 antibody was a significant prognostic factor for nodal metastasis of HNSCC. PMID- 11400230 TI - Low expression of p27(Kip1) in advanced mucoepidermoid carcinomas of head and neck. AB - BACKGROUND: p27(Kip1), a cyclin-dependent kinase inhibitor, negatively regulates the G1 phase progression of the cell cycle by binding to the cyclin E/cyclin dependent kinase 2 complex. This study was done to investigate the expression of p27(Kip1) in mucoepidermoid carcinomas and its usefulness as an indicator in tumor progression, aggressiveness, and prognosis. METHODS: Thirty-one patients with mucoepidermoid carcinomas who had surgical resection were studied retrospectively. Clinicopathologic features, including histologic types, T stage, nodal status, perineural invasion, overall AJCC stage, and survival data, were obtained from medical records. Immunohistochemical staining with monoclonal antibodies against p27(Kip1) was performed on the formalin-fixed, paraffin embedded specimens from each patient. The percentage of tumor cells expressing p27(Kip1) (labeling index) was evaluated by counting 1000 cells per slide in at least four different areas and comparing with the patients' clinicopathologic features and survival rates. RESULTS: Significant correlation was found between low p27(Kip1) expression and tumors with high-grade, advanced T stages, positive nodal status, and advanced clinical stages (p =.001 for all) except perineural invasion. Multivariate analysis indicated that p27(Kip1) expression (p =.030) was the most significant, and gender (p =.048) was the next significant predictor of overall survival among the variables. Also patients with low p27(Kip1) expression showed poor prognosis (p =.002). CONCLUSIONS: We suggest that p27(Kip1) is a reliable independent marker of tumor progression, invasiveness, and prognosis in the mucoepidermoid carcinomas. PMID- 11400231 TI - Survival in patients with recurrent squamous cell head and neck carcinoma treated with bio-chemotherapy. AB - BACKGROUND: We have shown that rIL-2 administration in recurrent head and neck cancers induces a tumor-specific T-lymphocyte reactivity and tumor regression; in a pilot study we have shown a safe and effective administration of rIL2 after cisplatin + 5-fluorouracil. Long-term results are not known. METHODS: Thirty patients with recurrent-persistent head and neck cancer were treated with cisplatin (100 mg-m(2)) d.1,5-fluorouracil (1 gr-m(2)-d c.i. 96 h), and SQ rIL-2 (4.5 M IU day 8 to 12 and 15 to 19) every 3 weeks. RESULTS: The overall response rate was 53.3% (95% CI; 34.4-72.3%): 26.6% complete response (CR) (8 patients) and 26.6% partial response (PR) (8 patients); 6 patients had SD (20%), 8 had PD (26.6%). The median follow-up was 36 months (range, 28-44). The median CR duration is 16.2 months (8.5-39+); the median survival duration of this group has not been reached. The median PR duration was 7.2 months (3-10); the median survival was 13.3 months (10-26). The median overall survival was 14 months. CONCLUSIONS: The most impressive finding is the very long survival of CRs patients. This outcome has been reported in other cancer patients with a CR after IL-2 therapy. PMID- 11400232 TI - Evaluation of thyroid nodules with technetium-99m MIBI and technetium-99m pertechnetate. AB - BACKGROUND: Most thyroid centers use fine-needle aspiration (FNA) and technetium 99m pertechnetate for the preoperative assessment of thyroid nodules. This approach is sufficient in most cases other than follicular neoplasm, and follicular carcinoma is more common than papillary carcinoma in developing countries such as in our center. Technetium 99m-methoxyisobutylisonitrile (MIBI) proposed for myocardial perfusion was also found to be taken up by a variety of tumors including thyroid cancer. METHODS: We evaluated MIBI uptake of nodular thyroid disease and compared it with pertechnetate scan, FNA, and histologic findings for the differentiation of malignant thyroid nodules from benign lesions. Seventy-one patients were included in the study. Three-phase pertechnetate scintigraphy was completed after a single injection of 150 MBq. Perfusion/uptake mismatch (uniform perfusion with cold uptake) was regarded as positive for malignancy, whereas perfusion/uptake match (cold perfusion with cold uptake) was regarded as negative. After 1 week, 400 MBq of MIBI was injected, images were obtained at 20 minutes and 2 hours, and evaluated semiquantitatively by use of a 4-point (0-3) scoring system. MIBI scans were considered positive if there was uptake superior to normal thyroid tissue on early and delayed images (score = 3). In the following days and weeks, all patients underwent FNA followed by surgery. RESULTS: Histopathologic diagnosis revealed a total of 23 thyroid carcinomas, 21 (91%) and 19 (83%) were positive on MIBI and pertechnetate, respectively. Of the 48 patients with benign nodules, 11 (23%) and 29 (60%) were positive on MIBI and pertechnetate, respectively. The specificity of MIBI, pertechnetate, and FNA is 77%, 40%, and 90%, respectively. CONCLUSIONS: In combination with FNA and three-phase pertechnetate scan, MIBI could be helpful in preoperative assessment of thyroid nodules. Intense MIBI activity increases the probability of thyroid cancer, whereas reduced activity drastically decreases the probability of malignancy. PMID- 11400233 TI - Surgical treatment of recurrent pleomorphic adenoma of the parotid gland: a clinical analysis of 52 patients. AB - BACKGROUND: Surgical management of recurrent pleomorphic adenoma of the parotid gland has a considerable risk of facial nerve injury and a high re-recurrence rate. To obtain more insight into this issue we evaluated our experiences. METHODS: Medical records and histologic material of all these patients (31 women and 21 men), who had been treated from 1976-1995 were reviewed. Median interval between initial treatment and commencement of recurrences was 3 (0.8-18) years. Last surgery consisted of parotidectomy in 48 patients (92%), including 19 (40%) total procedures and wide local excision with involved skin in four patients. RESULTS: At a median follow-up of 9 years, eight patients (8/52; 15%) had re recurrences develop, including 4 of 21 patients (19%) after a previous parotidectomy (group I) and 4 of 31 patients (13%) without prior parotidectomy (group II). The chance of re-recurrence in a group of patients with a minimal follow-up of 10 years after salvage surgery was 17% (4 of 24). The risk of a new relapse was, respectively, 4% and 8% at 1 and 5 years after treatment of recurrent disease. Acceptable N.VII function was preserved in 45 of the 49 (92%) nerves at risk. The risk of N.VII injuy was higher and more serious in group I (29% vs 10% in group II). The function of four of the five (80%) reconstructed zygomaticotemporal branches of the N.VII was adequate. CONCLUSION: Surgical treatment of recurrent pleomorphic adenoma of the parotid gland, usually consisting of a parotidectomy with wide extent and eventually facial nerve reconstruction, demonstrates favorable results with acceptable morbidity. The risk of new relapse and N.VII injury was higher after previous parotidectomy. PMID- 11400234 TI - Effects of xerostomia on perception and performance of swallow function. AB - BACKGROUND: Head and neck cancer treatment with high-dose chemoradiation may cause xerostomia and affect the patient's perception of swallowing ability. METHOD: Whole saliva production was measured in 36 patients with advanced-stage cancer of the oropharynx before treatment and 3 months after treatment by weighing a 4 x 4 inch gauze before and after a 2-minute chewing period. Presence of multiple eating difficulties was measured by patient interview. Swallowing was examined videofluorographically (VFG). RESULTS: Saliva weight decreased from a mean (SEM) of 5.1 (0.5) g pretreatment to 1.4 (0.5) g after treatment (p<.0001). At 3 months, significantly more patients perceived difficulty swallowing, dry mouth, needing water while eating, food stuck in the mouth or throat, and change in taste. Saliva weight was not correlated with VFG measures of bolus transit or observations of residue. CONCLUSIONS: Chemoradiation treatment results in xerostomia and a significant increase in patient perception of swallowing difficulties. Saliva weight in patients who perceive swallowing problems was lower. Xerostomia did not affect the physiologic aspects of bolus transport. Xerostomia affected the sensory process and comfort of eating more than bolus transport. PMID- 11400235 TI - Powered instrumentation for transcervical removal of gigantic intrathoracic thyroid. AB - BACKGROUND: It is widely accepted that almost all intrathoracic goiters can be removed through the neck. For those rare gigantic goiters that cannot be removed transcervically, median sternotomy is usually recommended. During the last 11 years we used intracapsular volume reduction techniques to facilitate transcervical removal of extremely large intrathoracic goiters. Materials and Methods Of 149 patients with intrathoracic goiters, 11 patients had gigantic lesions that could not be removed transcervically. Instead of sternotomy, we used the arthroscopic or sinus microdebrider or a large-bore suction device for controlled intracapsular volume reduction. This was followed by complete removal of the gland through the neck. RESULTS: The thyroid gland was removed completely in all 11 patients. None of the patients had any evidence of intraoperative spillage of thyroid tissue. No major complications were noted. CONCLUSIONS: We have found the use of the microdebrider and/or suction device for intracapsular volume reduction to be extremely helpful for transcervical removal of gigantic intrathoracic goiters. PMID- 11400236 TI - Lymphatic malformations of the head and neck: a retrospective review and a support for staging. AB - BACKGROUND: Lymphatic malformations (LM) are rare benign congenital tumors appearing mainly in the head and neck with a considerably variable outcome. A need exists to validate a staging system, taking into account the prognosis of the malformation, including preoperative and postoperative complications, long term sequelae, and persistence of the disease to improve parental counseling and evaluate the outcome of a surgical treatment of such tumors. METHODS: Twenty-two patients treated for LM were selected from a series of 129 patients operated on for congenital malformations of the head and neck between 1986 and 1997 at St-Luc University Hospital, Brussels, Belgium. Their charts were reviewed retrospectively, with a special focus on the anatomic location of the lesions and all the complications reported. According to de Serres et al, LM up to now have been divided into six possible categories according to their unilateral or bilateral infrahyoid and/or suprahyoid locations. RESULTS: Stage I (unilateral infrahyoid): nine patients, 11% of complications (without mediastinal extension: 0%, with mediastinal extension: 50%); stage II (unilateral suprahyoid): three patients, 33% of complications; stage III (unilateral suprahyoid and infrahyoid): eight patients, 75% of complications; stage V (bilateral suprahyoid and infrahyoid): two patients, 100% of complications. None of the children was initially seen with stage IV (bilateral suprahyoid) or stage VI (bilateral infrahyoid) LM. Overall complications, preoperative complications, postoperative com- plications, and long-term morbidity showed a significant in- crease from stage I to V (p <.01, p =.002, p =.02, and p =.03, respectively). CONCLUSIONS: A staging system of cervicofacial LM based on the anatomic location can be reliably used for prognostic purposes, allowing a more accurate assessment of the global risk of complications and determination of surgical outcome. Mediastinal extension in stage I patients seems to be associated with a higher rate of complications. Such information can be used to inform parents more appropriately regarding the management and long-term prognosis of their children's malformation. PMID- 11400237 TI - Sympathetic paraganglioma as an unusual cause of Horner's syndrome. AB - BACKGROUND: Paragangliomas are rare tumors arising from paraganglionic tissue of neural crest origin. They are present in any location where autonomic ganglia are found. The most common location in the head and neck is the carotid body, followed by the jugular bulb and vagus nerve. METHODS: A 30-year-old woman with a slowly growing left neck mass, aniscoria, and left eyelid ptosis was found to have a vascular tumor consistent with a paraganglioma arising near the left carotid bifurcation. After preoperative embolization, the patient underwent resection of the tumor. RESULTS: The tumor was found to be arising from the left sympathetic trunk and did not involve any other surrounding structures. Histopathologic analysis revealed the typical findings of a paraganglioma. CONCLUSIONS: Sympathetic paragangliomas are exceedingly rare tumors in the head and neck and should be considered in the differential diagnosis when clinical and radiographic evidence suggest a paraganglioma. The presentation is typically a slow-growing neck mass with the presence of an ipsilateral Horner's syndrome. PMID- 11400238 TI - Human dural replacement with acellular dermis: clinical results and a review of the literature. PMID- 11400239 TI - Surgical treatment of early oral carcinoma-results of a prospective controlled multicenter study. AB - PURPOSE OF THE STUDY: To assess whether early stage (pT1-2,pN0-1) oral cavity carcinoma is adequately treated by radical surgical resection alone. MATERIAL AND METHODS: Prospective multicenter study. Of 105 patients with cT1-2 cN0-1 oral carcinoma treated in conformity with the study design, 12 had to be excluded because of tumor-positive margins or pN stage > N1. The remaining 93 patients were monitored for at least 2 years. RESULTS: Seventeen patients had local or regional recurrence develop. In 12 of the 17 patients locoregional control was achieved by second treatment. Overall, the 4-year disease-specific survival probability was 94%. Patients treated initially without selective neck dissection had significantly higher recurrence rates than those with neck dissection, although the survival probability was not adversely affected. CONCLUSIONS: Early (pT1-2, pN0-1) squamous cell carcinoma of the oral cavity is adequately treated by surgery alone, provided the resection margins are tumor free. On the basis of the presented data, we would also advocate routine selective neck dissection. PMID- 11400240 TI - Prognosis of patients with recurrent laryngeal carcinoma. AB - BACKGROUND: Although recurrent laryngeal carcinoma is a common clinical problem, the data regarding its natural history and prognostic factors are limited. The aim of the study was to describe the clinical course of patients with recurrent laryngeal carcinoma and determine the impact of various factors on their survival. METHODS: The medical files of 65 patients with recurrent laryngeal cancer treated at the Rabin Medical Center between 1975 and 1996 were reviewed. The possible risk factors for survival were analyzed using univariate and multivariate models. RESULTS: The estimated 2- and 5-year survival rates of the patient population were 67% and 56%, respectively. On univariate analysis, primary tumor site, T stage and nodal status, duration of disease-free interval, site of recurrence, and operability of recurrent tumor were all powerful prognostic factors for survival. On multivariate analysis, three variables retained statistical significance: primary tumor site (p < .001), site of recurrence (p < .001), and its operability (p = .005). CONCLUSIONS: We found several disease-related factors to be predictive of poor outcome in patients with recurrent laryngeal cancer. Our data emphasize the need for early detection not only of primary tumors but also of their recurrences. PMID- 11400241 TI - Oral squamous cell carcinoma of the mandibular region: A survival study. AB - BACKGROUND: Oral squamous cell carcinomas (SCC) of the mandibular region present the lowest survival rates of the whole oral cavity. The purpose of this analysis was to evaluate the prognostic significance of several diagnostic and therapeutic variables in the survival rates of these carcinomas. METHODS: Forty-nine patients with oral cancers were treated by primary site surgery, involving mandibular resection in all cases. Thirty-one patients underwent postoperative adjuvant radiotherapy. RESULTS: Overall mean survival time and 5-year survival rate were 56.5 months and 44%, respectively. Patients in stages III and IV showed a statistically significant (p = .01) lower survival rate than those in stage II, and positive surgical margins had an adverse effect on survival (p = .03). No differences were found between patients treated by marginal or segmental mandibulectomy. CONCLUSIONS: Among the prognostic predictors studied, only the status of the surgical resection margin (odds ratio, 5.7) and tumor stage (III and IV vs II showed odds ratios of 2.1 and 3.6, respectively) affected the prognosis for SCC of the mandibular region. Tumor site was not associated with prognosis but was related with the probability that surgical margins were involved. PMID- 11400242 TI - Re-operation for bleeding after thyroidectomy and parathyroidectomy. AB - BACKGROUND: This study was undertaken to determine the frequency and timing of re operation for bleeding following thyroidectomy (THY) and parathyroidectomy (PARA) as well as the implications of this concerning the safety of ambulatory surgery. METHODS: Patients requiring re-operation after THY and PARA were identified from a computerized database of patients undergoing surgery between 3/l/95 and 12/31/99. The medical records of these patients were reviewed in detail. RESULTS: Six of 918 THY (0.7%) and 4 of 350 PARA (1.1%) required re-operation for bleeding. In two cases the wounds were opened emergently at the bedside due to worsening airway obstruction. One patient required an emergency tracheostomy. There were no deaths. Excluding one patient who bled five days post-operatively, the time interval from the completion of surgery to the identification of postoperative hematoma ranged from 2 to 48 hours, the median being 16 hours. CONCLUSIONS: Postoperative bleeding is an uncommon but unavoidable complication of THY and PARA. If treated promptly, serious consequences can be avoided. The relatively long interval between the initial operation and the development of the hematoma needs to be considered when recommending the performance of these procedures on an ambulatory basis. PMID- 11400243 TI - Localized non-Hodgkin's lymphoma of Waldeyer's ring: clinical features, management, and prognosis of 130 adult patients. AB - BACKGROUND: Waldeyer's ring (WR) is the primary site of non-Hodgkin's lymphoma (NHL) involvement in approximately 5% to 10% of all lymphoma patients, and it accounts for more than half of all primary extranodal lymphomas of the head and neck. Materials and Methods A retrospective review was performed of 130 adult patients with localized (stages I and II) WR-NHL seen at a single institution over 18 years. RESULTS: Patients had a median age of 55 years, and the male female ratio was 1:5:1. Seventy five (58%), 46 (35%), and 9 (7%) patients had primary tonsillar, nasopharyngeal, and base of the tongue lymphoma, respectively. Forty-five (35%) and 85 (65%) had stage I and stage II disease, respectively. Most patients (109 patients, 84%) had diffuse large B-cell NHL (DLC). Chemotherapy (CT) was given to 58 (45%) patients, whereas 26 (20%) received radiation therapy (RTX), and 46 (35%) were managed with a combination of chemotherapy and radiotherapy (CMT). One hundred nine (84%), 16 (12%), and 5 (4%) patients attained complete remission (CR), partial remission (PR), and treatment failure, respectively, with no difference in CR rates between the three therapeutic modalities. Of those patients with DLC, 90 (83%), 15 (14%), and 4 (3%) demonstrated CR, PR, and treatment failure, respectively. In a multivariate analysis, the modified International Prognostic Index (IPI) was found to predict the attainment of CR. Over a median follow-up of 49 months; 76 (58%) of the patients were alive and disease-free, 5 (4%) were alive with evidence of disease, and the remaining 49 (38%) were dead. Most distant relapses were in nongastrointestinal extranodal sites. The median overall survival (OS) has not been reached; however, the projected 5-year OS was 58%. No OS difference was noted between patients with stage I and stage II. Cox proportional hazards model identified primary tonsillar site and a low-risk group as defined by the modified IPI were associated with favorable OS. The median event-free survival was 82.3 months, with the primary tonsillar site, and low-risk modified IPI group were associated with favorable EFS in a multivariate analysis. Probably because of the high frequency of patients with DLC, the outcome and the prognostic factors in those patients were not distinctive from those for the whole group. The CMT was not associated with a superior OS compared with either of the single modality treatments; however, it was associated with more favorable EFS. CONCLUSIONS: This series characterized the clinicopathologic features and outcome of adult patients with early stage WR-NHLs. No survival difference was noted between stage I and stage II, and the outcome was favorable. Primary tonsillar site and the low-risk group of the modified IPI predicted favorable OS and EFS. CMT is probably superior to single modality treatment; however, prospective studies are warranted. PMID- 11400244 TI - Radioimmunotherapy in patients with head and neck squamous cell carcinoma: initial experience. AB - BACKGROUND: Despite improvements in locoregional treatment of stages III/IV squamous cell carcinoma of the head and neck (HNSCC), local and distant failure rates remain high. An effective adjuvant therapy is required for these patients. Among novel approaches is radioimmunotherapy, in which monoclonal antibodies (MAbs) are used for selective delivery of radiation to tumor cells. METHODS: The suitability of 186Re-labeled chimeric MAb U36 (186Re-cMAb U36) for radioimmunotherapy was evaluated in a phase I study, with radiation dose escalating steps of 11, 27, and 41 mCi/m2. Tumor targeting was monitored with a gamma camera, and the maximum tolerated dose was established in 13 patients with recurrent or metastatic disease. RESULTS: Administrations were well tolerated, and excellent targeting of tumor lesions was seen. Myelotoxicity was the only toxicity observed, resulting in dose-limiting toxicity in two patients treated with 41 mCi/m2. The MTD was established at 27 mCi/m2. A marked reduction in tumor size was observed in two patients, another showed stable disease for 6 months. CONCLUSIONS: Radioimmunotherapy with 186Re-cMAb U36 seems to be well tolerated, with bone marrow being the dose-limiting organ. The observation of antitumor effects is encouraging for further development of radioimmunotherapy for HNSCC. PMID- 11400245 TI - The significance of comorbidity in advanced laryngeal cancer. AB - INTRODUCTION: Cancer patients often have concurrent diseases and conditions known as comorbidities. The aim of this project is to demonstrate the significance of comorbidity in the treatment and outcomes of advanced laryngeal carcinoma. METHODS: A retrospective medical record review of 182 patients with previously untreated T3 or T4 squamous carcinomas of the larynx treated at M. D. Anderson between 1990 and 1995 was performed. Demographic, patient-specific, tumor specific, and outcome measures information were collected. Comorbidity was coded using the Modified Medical Comorbidity Index. Univariate and multivariate analysis with the use of life survival analysis techniques and logistic regression were performed. RESULTS: The median age at diagnosis was 59.5 years. Most patients were men (69.2%) and Caucasian (73.1%). Laryngeal preservation was performed in 90 patients, and surgical resection was performed in 92 patients. Patients in the two treatment groups had similar comorbidity, locoregional control (65%), and 5-year survival (37.3%). Patients with either moderate or severe comorbidity had significantly worse overall survival (p = .00014) and worse 5-year survival than those with no or mild comorbidity (21.8% vs 46.3%, p = .003). CONCLUSIONS: This study demonstrates that comorbidity is significantly associated with survival in a group of patients with identical histology, site, and stage. Comorbid status should be incorporated into the assessment of prognosis and outcome to improve and optimize the management of head and neck cancer patients. PMID- 11400246 TI - Cytoplasmic accumulation of alpha-catenin in thyroid neoplasms. AB - BACKGROUND: Alpha-catenin (alpha-cat) is one of the anchoring proteins of E cadherin. It has been shown that deviation in its function may alter the cadherin catenin complex leading to disturbed cell-cell adhesion. Immunohistochemical studies have shown that cytoplasmic localization of alpha-cat in tumors is associated with aggressive behavior. In this study, we evaluated the expression of alpha-cat in various thyroid tumors by immunohistochemical analysis. METHODS: Fifty cases were selected:18 papillary carcinoma classic type (PTC), 9 follicular variant of PTC (FVPTC), 6 follicular carcinoma (FCA), 4 anaplastic carcinoma (ACA), 8 follicular adenoma (FA), 3 nodular goiter (NG), and 2 lymphocytic thyroiditis (LT). The staining reaction was classified as membranous, cytoplasmic, or both. The intensity of the staining was graded as negative (0), weak (+), moderate (++), and strong (+++). RESULTS: Staining along the cell membrane was observed in 36 (72%) and cytoplasmic expression was present in 28 (57%) cases. The cytoplasmic staining was more commonly seen in malignant lesions; it was more common in PTC (78% of all PTC) than follicular patterned lesions (FVPTC, FCA). All cases of ACA (4 of 4) showed only cytoplasmic expression. No correlation was found between lymph node involvement and alpha-cat staining patterns. CONCLUSIONS: Cytoplasmic expression of alpha-cat is more common in (1) malignant lesions of thyroid and (2) PTC than FVPTC and FCA. The lack of membrane and presence of cytoplasmic expression suggest a role of alpha catenin in the aggressive biology of ACA. PMID- 11400247 TI - Choosing a concomitant chemotherapy and radiotherapy regimen for squamous cell head and neck cancer: A systematic review of the published literature with subgroup analysis. AB - BACKGROUND: A systematic review was conducted to develop clinical recommendations for concomitant chemotherapy (CT) and radiotherapy (RT) in patients with locally advanced squamous cell head and neck cancer (SCHNC). METHODS: Results of published randomized controlled trials (RCTs) were pooled using Meta analyst(0.988) software. RESULTS: A pooled analysis of 18 RCTs (20 comparisons) involving 3,192 patients detected a reduction in mortality for concomitant therapy compared with RT alone (odds ratio [OR], 0.62; 95% confidence interval [CI], 0.52-0.74; relative risk, 0.83; risk reduction, 11%; p < .00001). Platinum based regimens involving 1,514 patients from nine trials (10 comparisons) were most effective (OR, 0.57; 95% CI, 0.46-0.71; p < .00001; risk reduction, 12%). Concomitant therapy produced more acute adverse effects than RT alone. CONCLUSION: Platinum-based concomitant CT and RT is superior to conventional RT alone in improving survival in locally advanced SCHNC. Subgroup analyses can be used to help in choosing the most appropriate concomitant regimen. PMID- 11400248 TI - Lateral cervical bronchogenic cyst: an unusual cause of a lump in the neck. AB - BACKGROUND: Congenital cysts of the neck in children are not uncommon. Most of these are thyroglossal, branchial cleft, or less commonly, thymic cysts. Bronchial cysts rarely are initially seen as a neck mass. METHODS: Use of an illustrative case of a bronchogenic cyst initially seen as an upper lateral neck mass. CONCLUSIONS: We emphasize that although ectopic bronchogenic cysts are rare lesions of the head and neck, especially in the upper lateral neck, they should be included in the differential diagnosis in the evaluation of congenital neck cysts. PMID- 11400249 TI - Cutaneous botryomycosis of the cervicofacial region. AB - BACKGROUND: Botryomycosis is a rare, chronic, bacterial infection of insidious onset involving the integument or viscera that often mimics actinomycosis or a deep fungal infection. The pathogenesis is thought to be a symbiotic relationship between the host and the infecting organism. METHODS: Case report of a patient with a chronic infection involving the cervicofacial region diagnosed as cutaneous botryomycosis arising from a chronic osteomyelitis of the mandible. The diagnosis was based on the chronicity of the infection along with the identification of botryomycotic (bacteria-containing) granules on histopathologic examination. Special stains excluded fungi and mycobacterium. Cultures identified the offending bacteria, and antibiotic therapy was initiated on the basis of the sensitivities, resulting in resolution of this chronic infectious process. A review of the English language literature revealed that this is the first case of cutaneous botryomycosis arising from a chronic osteomyelitis of the mandible. RESULTS: Medical therapy proved curative at 14 months follow-up. Surgery was performed for diagnostic purposes only. CONCLUSIONS: Botryomycosis is exceedingly rare in the head and neck, and consideration of this entity in the differential diagnosis is critical to the diagnosis. The mainstay of therapy is medical with surgery reserved for biopsy and/or excision of persistent disease. Published 2001 John Wiley & Sons, Inc. PMID- 11400250 TI - Science or marketing at WHO? A response to Williams. PMID- 11400252 TI - The role of permanent income and family structure in the determination of child health in Canada. AB - We use data from the Ontario Child Health Study (OCHS) to provide the first Canadian estimates of how the empirical association between child health and both low-income and family status (lone-mother versus two-parent) changes when we re estimate the model with pooled data. Two waves of data provide a better indication of the family's long-run level of economic resources than does one wave. Our measures of health status include categorical indicators and the health utility score derived from the Health Utilities Index Mark 2 (HUI2) system. Consistent with findings from other countries, we find that most outcomes are more strongly related to low-average income (in 1982 and 1986) than to low current income in either year. Unlike some previous research, we find the quantitative impact of low-income on child health to be modest to large. Lone mother status is negatively associated with most outcomes, but the lone-mother coefficients did not change significantly when we switched from low-current income to low-average income. This implies that the lone-mother coefficient in single cross-sections is not just a proxy for low-permanent income. PMID- 11400253 TI - A framework for cost-effectiveness analysis from clinical trial data. AB - We present a general Bayesian framework for cost-effectiveness analysis (CEA) from clinical trial data. This framework allows for very flexible modelling of both cost and efficacy related trial data. A common CEA technique is established for this wide class of models through linking mean efficacy and mean cost to the parameters of any given model. Examples are given in which efficacy may be measured as a continuous, binary, ordinal or time-to-event outcome, and in which costs are modelled as distributed normally, log-normally, as a mixture or non parametrically. A case study is presented, illustrating the methodology and illuminating the role of prior information. PMID- 11400254 TI - Recognizing diversity in public preferences: the use of preference sub-groups in cost-effectiveness analysis. AB - Public preferences are typically incorporated into cost-effectiveness analyses (CEA) on the basis of the average health state utilities of a sample of individuals drawn from the general public. The cost-effectiveness of a programme is then assessed on an 'all-or-nothing' basis: the programme is declared either cost-effective or not for all patients in clinically homogeneous sub-groups. However, this approach fails to recognize variability between individuals in their preferences. In this paper, we consider how diversity in the preferences of individuals can be handled within CEA when the public's preferences are considered appropriate for defining benefit, with the objective of increasing the efficiency of health care delivery. The concept of preference sub-group analysis is described and some of its implications are assessed. These include the methods that could be used to identify sub-groups from amongst public raters, the appropriate approach to eliciting preferences and the possible implications of preference sub-group analysis for clinical decision making. PMID- 11400255 TI - Provider payment reform in China: the case of hospital reimbursement in Hainan province. AB - This paper develops a simple model of payment incentives and empirically evaluates provider payment reform in Hainan Province, China. We use a pre-post study design with a control group to analyse two years of claims data to assess the impact of a January 1997 change to prospective payment for a sub-sample of the hospitals. This difference-in-difference empirical strategy allows us to isolate the supply-side payment reform effects from demand-side changes, in contrast with previous studies of China's reforms. Our results validate the theory that Chinese providers' behavioural response to payment incentives is similar to that reported in the literature derived from the experience of industrialized countries. We find that prepayment is associated with a slower rate of growth of overall expenditures, programme spending and patient co payments per inpatient admission, compared to fee-for-service (FFS). These findings suggest cautious optimism regarding the effectiveness of prospective payment for controlling costs and should be encouraging for policymakers in developing and transitional economies considering replacement of FFS with more aggregated forms of provider payment. PMID- 11400256 TI - The effect of health on acute care supplemental insurance ownership: an empirical analysis. AB - The empirical effect of health status on private insurance ownership is a mixture of the effect of health on the demand for insurance (subjected to adverse selection) and its effect on the insurer's underwriting practice (subjected to risk-selection). Using bivariate partial observability probit models, this paper provides an empirical identification of health effects on the probability of application and on the probability of rejection in the Israeli market for acute care supplemental health insurance. The analysis shows that while the reduced form health effect on ownership is negligible, the structural effects are sizeable and indicate that sicker individuals are more likely to apply, but are also more likely to be rejected. The policy implications of the above findings are discussed in the context of the Israeli health system. PMID- 11400257 TI - Private and social time preferences for health and money: an empirical estimation. AB - The main objective of this paper is to examine the relationship between individual time preference for health and money. To that end, we tested whether individuals discount their own health at the same rate as their own money and, similarly, whether they discount social health in the same terms as social money. To offer private and social money and health choices is, to the best of our knowledge, new in the literature on the estimation of time preferences and, in our view, represents a valid way in which to respond to the question of the uniform or differential discount of health, as against monetary, consequences. The results of our estimation suggest that a sample of students exhibit higher time preference rates for health than money, this being the case when the exchanges are both private and public. PMID- 11400258 TI - The impact of income inequality on individual and societal health: absolute income, relative income and statistical artefacts. AB - The relative income hypothesis, that relative income has a direct effect on individual health, has become an important part of the literature on health inequalities. This paper presents a four-quadrant diagram, which shows the effect of income, relative income and aggregation bias on individual and societal health. The model predicts that increased income inequality reduces average health regardless of whether relative income affects individual health. If relative income does have a direct effect then societal health will decrease further. PMID- 11400259 TI - Cost prediction models for the comparison of two groups. AB - For trial-based economic evaluation where patient-specific cost data are not routinely available, cost prediction models are commonly used to estimate total cost for each patient. Typically, multiple regression techniques are used on data from diagnosis-matched, non-trial patients (where patient-level cost data are available) to model cost as a function of covariates that are observed on the trial subjects (e.g. length of hospital stay, procedures, etc.). The estimated beta coefficients provide a means of estimating the total cost for each patient in the trial. However, the variability of the beta coefficients due the measurement and sampling error is seldom included in the overall variance expression for mean costs by treatment group. In this paper we provide a method for estimating this variance and provide an example application PMID- 11400260 TI - Flow-chemiluminescence: a growing modality of pharmaceutical analysis. AB - The present article deals with a review of the applications of the chemiluminescence to pharmaceutical analyses by using continuous-flow assemblies. A comprehensive study on the fundamentals of the chemiluminescence is also included. PMID- 11400261 TI - Screening of chemiluminescence constituents of cereals and DPPH radical scavenging activity of gamma-oryzanol. AB - The chemiluminescence (CL) constituents of cereals were detected by CL using the H(2)O(2)-acetaldehyde system. The cereals tested, such as rice, millet and sorghum, exhibited various levels of CL activity. The gamma-oryzanol fraction was extracted from brown rice and separated into four constituents by HPLC. The four constituents were identified as cycloartenyl ferulate, 24-methylenecycroartanyl ferulate, campesteryl ferulate and beta-sitosteryl ferulate. Free radical scavenging activities with 1,1-diphenyl-2-picrylhydrazyl (DPPH) and CL intensities of four constituents (gamma-oryzanol components) were measured and compared with that of gallic acid, which is a typical free radical scavenger. Four constituents scavenged DPPH radicals and scavenging activities were proportional to CL intensities. Concentrations of four CL constituents required to quench 50% (IC(50)) of the free radicals ranged from 0.9 to 1.1 mmol/L. We demonstrated that measurement of CL intensities was a rapid and convenient method for screening DPPH radical scavenging activities of rice. PMID- 11400262 TI - Upper electron-excited states in bioluminescence: experimental indication. AB - The involvement of upper electron-excited states in bacterial bioluminescence process was studied with excitation energy-accepting molecules. The fluorescent aromatic compounds, anthracene and 1.4-bis(5-phenyloxazol-2-yl)benzene, were chosen. Energies of their lowest excited singlet states are higher than the energy of the analogous state of the bioluminescence emitter; their absorption spectra and bioluminescence do not overlap. Hence, the excitation of these molecules by singlet-singlet energy transfer or by light absorption is excluded. Sensitized fluorescence of these compounds in the bioluminescence systems has been recorded, indicating the activity of upper electron-excited states in the bioluminescent process. PMID- 11400263 TI - Detection of Texas red-labelled double-stranded DNA by non-enzymatic peroxyoxalate chemiluminescence. AB - We have found previously that different fluorescent dyes cannot be efficiently excited by the bis(2,4,6-trichlorophenyl)oxalate (TCPO)-H(2)O(2) reaction when they are intercalated between the DNA bases or bound to the minor groove of the double helix. Here we show that the fluorescent dye Texas red, covalently bound to the 3' ends of double-stranded DNA molecules, exhibits a high emission intensity when excited by the TCPO-H(2)O(2) reaction. In this case, the charge transfer between the intermediate produced in the peroxyoxalate chemiluminescent reaction and Texas red can take place because this fluorophore is not buried inside the DNA structure. We describe the application of this chemiluminescent reaction to the detection of blotted DNA on nylon membranes. PMID- 11400264 TI - Increasing the specificity of the forensic luminol test for blood. AB - It is shown that the presumptive luminol chemiluminescence test for the presence of traces of blood can be made more determinative by measuring the peak emission wavelength of the luminol chemiluminescence. When sprayed onto a surface containing traces of human haemoglobin, a 1 g/L solution of aqueous luminol containing 7 g/L sodium perborate gives an emission peak at 455 +/- 2 nm, whereas the same mixture gives an emission peak at 430 +/- 3 nm when sprayed onto a surface containing traces of sodium hypochlorite (household bleach). This spectral difference can readily be determined using spectroscopic equipment that either scans the spectrum before significant luminescence decay occurs or corrects the spectrum for the effects of any decay. It was found that bovine haemoglobin and human haemoglobin showed no significant spectral differences. PMID- 11400265 TI - A message to the scientific community. PMID- 11400266 TI - [Immuno-inflammatory responses and upper gastrointestinal disorders: immuno inflammatory mechanisms of chronic active gastritis]. PMID- 11400267 TI - [Gastrointestinal manifestations in collagen disease]. PMID- 11400268 TI - [The pathogenesis of inflammatory bowel disease viewed from immunological aspects]. PMID- 11400269 TI - [A case of ulcerative colitis along with characteristic features on computed tomography (CT), developed by the treatment with interferon for chronic hepatitis C]. PMID- 11400270 TI - [A case of so-called hamartomatous polyp of the colon]. PMID- 11400271 TI - [A case of multiple liver metastases from rectal carcinoid tumor successfully treated with arterial infusion chemotherapy using degradable starch microspheres (DSM)]. PMID- 11400272 TI - [A case of drug induced liver injury caused by a herbal drug, bofu-tsu-sho-san]. PMID- 11400273 TI - [A case of fulminant hepatitis, possibly caused by benzbromarone]. PMID- 11400274 TI - [A case of acute emphysematous cholecystitis accompanied with subphrenic abscess]. PMID- 11400275 TI - [A case of osteoclastoid type giant cell carcinoma of the pancreas]. PMID- 11400276 TI - [A case of inflammatory fibrosarcoma of the retroperitoneum]. PMID- 11400277 TI - [A case of pathogenic Escherichia coli O157 enteritis: usefulness of abdominal ultrasonography and serological test]. PMID- 11400278 TI - [Chronic hepatitis B]. PMID- 11400279 TI - [Antiviral therapy for patients with chronic hepatitis C]. PMID- 11400280 TI - [Colitis enhances the colorectal carcinogenesis in rats: correlation between the incidence of aberrant crypt foci and the incidence of tumors]. AB - We examined the implication of colitis on the colorectal carcinogenesis in rats. We used 1, 2-dimethylhydrazine (DMH) as a carcinogen and trinitrobenzenesulfonic acid (TNB) as a colitis-inducing agent on F344 rats. After treating the rats with DMH, TNB markedly enhanced the incidence of aberrant crypt foci (ACF), putative preneoplastic lesions, as well as colon cancers in the rats (p < 0.01). There was positive correlation between the incidence of ACF and the incidence of tumors. Furthermore, we treated the rats with two different anti-inflammatory drugs (a non-steroidal anti-inflammatory drug: Fenbufen and a platelet activating factor receptor antagonist: PAF-RA) after pre-treatment with DMH and TNB. Only PAF-RA significantly decreased the incidence of ACF in the rats (p < 0.05). PMID- 11400281 TI - [A case of amyloidosis of the large intestine which showed peculiar appearance and caused hematochezia]. PMID- 11400282 TI - [A case of nodular amyloidosis of the small intestine associated with small intestinal cancer]. PMID- 11400283 TI - [A case of ulcerative colitis complicated with rectovaginal fistula causing perianal abscess]. PMID- 11400284 TI - [A case of ileal heterotopic pancreas causing melena]. PMID- 11400285 TI - [A case of tuberculosis of the small intestine associated with panperitonitis due to intestinal perforation]. PMID- 11400286 TI - [A case of successful treatment of type II hepatorenal syndrome with transjugular intrahepatic portosystemic shunt using carbon dioxide as a contrast medium]. PMID- 11400287 TI - [A case of cryoglobulinemic membranoproliferative glomerulonephritis associated with chronic hepatitis C which was effectively treated by interferon alpha-2b]. PMID- 11400288 TI - [Bleeding gastric varices associated with pancreatic arteriovenous malformation]. PMID- 11400289 TI - [Use of transgenic and gene-targeted mice for research on hematological disorders]. PMID- 11400290 TI - The porphyrias: genetic and acquired aspects. PMID- 11400291 TI - [The mechanism of cell cycle regulation and its clinical relevance]. PMID- 11400292 TI - [Clinical implication of human dendritic cells]. PMID- 11400293 TI - [Role of chemokines in the development of autoimmune diseases]. PMID- 11400294 TI - [Role of a chemokine SDF-1/PBSF and its receptor CXCR4 in hematopoiesis]. PMID- 11400295 TI - [HIV-1 cell entry mediated by chemokine receptors]. PMID- 11400296 TI - [Chemokines in migration of ATL cells]. PMID- 11400297 TI - [Strategy for treatment of malignant lymphoma: from the standpoint of pathology]. PMID- 11400298 TI - [Mantle cell lymphoma]. PMID- 11400299 TI - [Treatment strategy for indolent B-cell lymphoma]. PMID- 11400300 TI - [High-risk aggressive lymphoma (ATL)]. PMID- 11400301 TI - [The role of high-dose chemotherapy with hematopoietic stem cell transplantation for malignant lymphoma]. PMID- 11400302 TI - [Pure red cell aplasia developing during the course of primary macroglobulinemia associated with pseudochylothorax and IgA nephropathy]. AB - A 74-year-old man who had undergone artificial pneumothorax therapy for pulmonary tuberculosis 40 years earlier was admitted because of blurred vision, headache, and numbness of the lower limbs in October 1997. He presented with anemia and leukocytopenia with monoclonal gammopathy of IgM (kappa). His bone marrow was diffusely infiltrated with small lymphocytes, plasmacytoid lymphocytes, and plasma cells expressing IgM, kappa surface immunoglobulin. On the basis of these findings, primary macroglobulinemia (PMG) was diagnosed. CT scan of the chest demonstrated pleural effusion of the right lung encapsulated in a thickened pleura, and pseudochylothorax was diagnosed from a specimen of chyliform fluid which contained numerous cholesterol crystals and was positive for Mycobacterium tuberculosis (MT) on PCR assay. The patient's condition was also complicated by chronic renal failure due to IgA nephropathy, which may have been a consequence of the tuberculosis, possibly due to an abnormal IgA-mediated immune response to MT. The patient gradually developed pure red cell aplasia during the course, probably due to an autoimmune mechanism. Later in the course, immunoglobulin gene analysis of the malignant cells of PMG showed that they were derived from antigenically selected cells. In the context of antigenic stimulation, the role of MT antigen in the pathogenesis of PMG was of interest in this patient. PMID- 11400303 TI - [CD13- and CD33-negative acute myelocytic leukemia (FAB classification; M2) with morphological changes and CD13 expression on recurrence]. AB - A 42-year-old man was diagnosed as having acute myelocytic leukemia in July 1998. The leukemic cells tended to be differentiated, and on the basis of positive peroxidase staining, this case was considered to be AML (M2) according to the FAB classification. t(8;21)(q22;q22) chromosomal abnormality was observed, but surface antigen analysis revealed no expression of either CD13 or CD33, a finding characteristic of myelocytic leukemia. Combination chemotherapy resulted in complete remission, and allogeneic bone marrow transplantation was performed with donor cells from the patient's sister. Unfortunately, however, the patient died about 18 months after the onset of leukemia. Comparison of the findings at recurrence with those at initial diagnosis revealed morphological changes in non differentiated immature cells (AML-M1) and CD13 surface antigen expression. This was considered to be a rare case of AML with neither CD13 nor CD33 expression at onset, but with CD13 expression at recurrence. PMID- 11400304 TI - [Transient nephrotic syndrome after allogeneic bone marrow transplantation for chronic myelogenous leukemia]. AB - A 42-year-old man with chronic myelogenous leukemia underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) from an unrelated donor in January 1998. About 100 days later, he developed skin eruption and a diagnosis of chronic graft-versus-host disease (cGVHD) was made by skin biopsy. The eruption improved with steroid therapy, and the dose of steroid was gradually tapered. On day 151, the patient developed nephrotic syndrome with proteinuria up to 20 g/day. A renal biopsy carried out on day 160 showed minimal change in the glomeruli. The proteinuria disappeared 19 days after the onset of nephrotic syndrome without any additional therapy, and no recurrence was observed upon re tapering of the steroid. In this case, cGVHD might have been related to development of the nephrotic syndrome. Nephrotic syndrome after allo-HSCT is a rare complication, and only ten cases have been reported. The histological findings were mainly membranous nephropathy, and immunosuppressive therapy was effective. As seen in this case, transient nephrotic syndrome with cGVHD may occur after allo-HSCT, and care is necessary to ensure that treatment of cGVHD is sufficient. PMID- 11400305 TI - [Simultaneous occurrence of Basedow's disease and autoimmune hemolytic anemia]. AB - A 43-year-old man with a 7-year history of low antinuclear factor titer developed Basedow's disease and autoimmune hemolytic anemia (AIHA) simultaneously. Such simultaneous occurrence of these autoimmune disorders has been reported only rarely. Administration of methimazole for Basedow's disease and prednisolone for AIHA was effective for ameliorating both conditions. The patient had the HLA DR2, DRB1 1501 and DPB1 0501 alleles, suggesting a genetic predisposition for these diseases. PMID- 11400306 TI - [Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (1)--Single oral and intravenous dose toxicity studies in rats]. AB - A single oral dose toxicity study of Cefmatilen hydrochloride hydrate (S-1090) and a single intravenous dose toxicity study of its sodium salt (S-1090-Na) were conducted in rats. One dose level of 2000 mg potency/kg was set in both studies. Single oral dose toxicity study of S-1090 No deaths occurred. Diarrhea occurred on the dosing day and slightly soft feces lasted until 6 days after administration. These changes were considered to result from changes of intestinal flora induced by the antibiotic activity of S-1090. Reddish-brown feces (due to chelated products of S-1090 or its decomposition products with Fe3+ in the diet) were also observed until the next day after administration. Body weights increased favorably, and no S-1090-related pathological changes were observed. The oral lethal dose of S-1090 was estimated to be more than 2000 mg potency/kg. Single intravenous dose toxicity study of S-1090-Na No deaths occurred. The rats showed characteristic clinical signs such as hypoactivity, abnormal gait and hypopnea immediately after dosing, and some rats showed prone position or paleness of eyeballs and ear auricles in due course. These signs disappeared by 4 hr after administration. Slightly soft feces and reddish-brown feces were observed much the same as in the orally-treated rats. Body weights increased favorably. In the pathological examinations, slight cecal enlargement and increased basophilia, dilatation and calcification of the renal tubules in the kidney were observed. The intravenous lethal dose of S-1090-Na was estimated to be more than 2000 mg potency/kg. PMID- 11400307 TI - [Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (6)--Six-month repeated oral dose toxicity study in dogs]. AB - A six-month repeated oral dose toxicity study of Cefmatilen hydrochloride hydrate (S-1090) at dose levels of 40, 100 and 250 mg potency/kg/day was conducted in male and female beagle dogs. No toxicologically significant changes were observed in general conditions of all animals. Reddish-brown feces (due to chelated products of S-1090 or its decomposition products with Fe3+ in the diet) were observed in all treated groups. Plasma irons showed a tendency to increase in the males and females of the 250 mg potency/kg group. However, as no changes suggesting anemia or hepatic injury were observed in this group, the change of plasma iron was considered to have no toxicological significance. No toxicologically significant changes were observed in other examination items. The plasma S-1090 concentrations increased in a manner less than dose-proportional. Based on the above results, the NOAEL of S-1090 was assessed to be 250 mg potency/kg/day. PMID- 11400308 TI - [Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (3)--One- and three month repeated oral dose toxicity studies in rats]. AB - One- or three-month repeated oral dose toxicity studies of Cefmatilen hydrochloride hydrate (S-1090) in rats were conducted. Doses were set at 80, 200, 500 and 1250 mg potency/kg/day in the one-month toxicity study, and 100, 300 and 1000 mg potency/kg/day in the three-month toxicity study. Body weights increased favorably and no deaths occurred in all treated groups of both studies. The changes observed in both studies were soft feces, abdominal distention, increased food and water consumption, decreases of urine volume and pH, and a decrease of blood neutrophils in almost all treated groups, reddish-brown feces (due to chelated products of S-1090 and its decomposition products with Fe3+ in the diet) in groups dosed at 300 mg potency/kg or more, and a lower mature granulocyte ratio in the bone marrow in groups dosed at 1000 mg potency/kg or more. In necropsy, cecal enlargement with a large amount of muddy content was observed in all treated groups of both studies. In the three-month toxicity study, elevated drug-metabolizing enzyme activities were noted in the liver of the males in the 1000 mg potency/kg group. These changes were slight except for the cecal enlargement and the rats recovered well with drug withdrawal. Since no toxicologically significant changes were noted in either study, the NOAEL of S 1090 was estimated to be 1250 mg potency/kg/day in the one-month toxicity study and 1000 mg potency/kg/day in the three-month toxicity study. PMID- 11400309 TI - [Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (7)--Three-month repeated oral dose toxicity study in juvenile dogs]. AB - To evaluate the repeated oral dose toxicity of Cefmatilen hydrochloride hydrate (S-1090) in juvenile dogs, S-1090 was administered to juvenile beagle dogs at dose levels of 50, 100, 200 and 400 mg potency/kg/day for 3 months. No deaths occurred. Urinalysis in the 400 mg potency/kg group revealed positive reactions of occult blood and protein, and erythrocytes in sediments. Cystitis was observed in the 200 and 400 mg potency/kg groups. In the thyroids, an increased weight in some animals in the groups dosed at 100 mg potency/kg or more and an increased follicular colloid in the 400 mg potency/kg group were observed. However, no related changes were noted in other examination items. Red to dark-red feces (due to chelated products of S-1090 or its decomposition products with Fe3+ in the diet) were observed in all treated groups. Plasma S-1090 concentrations increased in a manner less than dose-proportional. The lesions of urinary bladder were judged as S-1090-induced toxic changes and the NOAEL of S-1090 in this study was assessed to be 100 mg potency/kg/day. PMID- 11400310 TI - [Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (8)--Nephrotoxicity study in rabbits by single oral administration]. AB - A nephrotoxicity study of Cefmatilen hydrochloride hydrate (S-1090) was conducted in rabbits at single oral doses of 250, 500 and 1000 mg potency/kg. All treated groups showed a decreased food consumption and a tendency for the body weight to decrease. Urinary protein and glucose were detected and slight increases of plasma creatinine and urea nitrogen were observed in the 500 mg potency/kg group. Urinary protein was also detected in the 1000 mg potency/kg group. In the histopathological examination of the kidney, tubular necrosis was observed in the 500 and 1000 mg potency/kg groups. No nephrotoxic signs were observed in the 250 mg potency/kg group. The NOAEL on the nephrotoxicity of S-1090 in rabbits was estimated to be 250 mg potency/kg. PMID- 11400311 TI - [Reproductive and developmental toxicity studies of S-1090, cefmatilen hydrochloride hydrate (1)--A study on oral administration prior to and in the early stages of pregnancy in rats]. AB - Cefmatilen hydrochloride hydrate (S-1090) was administered daily by gavage to rats at doses of 100, 300 or 1000 mg potency/kg/day prior to and in the early stage of pregnancy to assess its adverse effects on parental reproductive ability and embryo-fetal development. Loose and/or reddish brown feces were observed in both males and females of all the S-1090 dosing groups, and abdominal distention was also observed in males throughout the dosing period. No drug-related deaths occurred in either males or females. In males, body weight and food consumption were increased at a dose of 1000 mg potency/kg/day throughout the dosing period. In females, body weight gain was restrained during late pregnancy, and food consumption was decreased transiently following the initiation of dosing, and then remained high on the day before parturition in all the S-1090 dosing groups. Necropsy of male and female rats revealed an increase in the cecum weight. The reproductive ability of males and females was normal in all the S-1090 dosing groups. No effects of S-1090 were observed in the implantation ratio, embryo fetal viability, fetal body weight, and incidence of external, skeletal and visceral anomalies. Based on these results, the no observed adverse effect levels of S-1090 are estimated to be less than 100 mg potency/kg/day for parental general toxicity, 1000 mg potency/kg/day for reproductive toxicity, and 1000 mg potency/kg/day for developmental toxicity in embryo-fetuses under the conditions of the present study. PMID- 11400312 TI - [Reproductive and developmental toxicity studies of S-1090, cefmatilen hydrochloride hydrate (2)--A study on oral administration during the period of organogenesis in rats]. AB - Cefmatilen hydrochloride hydrate (S-1090) was administered daily by gavage to female rats at doses of 100, 300 or 1000 mg potency/kg/day from Days 7 to 17 of pregnancy to assess its effects on dams and on development of the embryo-fetuses and offspring. Loose or reddish-brown feces were observed in dams of all the S 1090 dosing groups. Body weight gain was increased from the early stage of administration to the end of pregnancy, food consumption was transiently decreased at the early stage of administration, and water consumption was increased from the middle to the end of pregnancy in all the S-1090 dosing groups. However, no effects on pregnancy, parturition and lactation were observed. Necrospy revealed an increased cecum weight in pregnant and lactating dams of all the S-1090 dosing groups. No effects of S-1090 were observed in viability, growth, incidences of external, skeletal and visceral anomalies, and degree of ossification in F1 fetuses. No effects of S-1090 were observed in such parameters as viability, incidence of external and skeletal anomalies, physical development, sensory functions/reflexes, behavior and reproductive function in F1 offspring. No adverse effects were observed in F2 offspring. On the basis of these results, the no observed adverse effect levels of S-1090 are estimated to be less than 100 mg potency/kg/day for maternal general toxicity, 1000 mg potency/kg/day for maternal reproductive toxicity and the developmental toxicity in the embryo-fetuses and offspring under the conditions of the present study. PMID- 11400313 TI - [Reproductive and developmental toxicity study of S-1090, cefmatilen hydrochloride hydrate (3)--A study on oral administration during the period of organogenesis in rabbits]. AB - Cefmatilen hydrochloride hydrate (S-1090) at dosage levels of 6.25, 12.5 and 25 mg potency/kg/day was administered orally by gavage to groups of 13-16 pregnant rabbits daily during the period of organogenesis, and the effects of S-1090 on dams and fetuses were examined. Control animals were treated with a 0.5 w/v% aqueous solution of methylcellulose. Abortion was noted in 1 of the 16 females of the 12.5 mg potency/kg group and in 1 of the 14 females of the 25 mg potency/kg group, and death was noted in 1 of the 14 females of the 25 mg potency/kg group. Regarding the dams, decreased food consumption was noted in the 12.5 mg potency/kg group in the beginning of the dosing period. Suppressed body weight gain and decreased food consumption were noted in the 25 mg potency/kg group during the pregnancy period. At necropsy, thickening of the gastric mucosa, hemorrhage in the cecum, and higher values of cecum weight were also noted in this group. On the other hand, no effects of S-1090 were noted in general signs, body weight changes, food consumption, necropsy findings, or organ weights in the 6.25 mg potency/kg group. No effects of S-1090 were noted in the number of corpora lutea, number of implantations, implantation rate, death or resorption rate, number of live fetuses, sex ratio of live fetuses, fetal body weight of either sex, incidence of external anomalies, incidence of skeletal anomalies or variations, degree of ossification, or incidence of visceral anomalies. On the basis of the above results, the no observed adverse effect levels of S-1090 are estimated at 6.25 mg potency/kg/day for general toxicity and reproductive functions in dams, and at 25 mg potency/kg/day for development in fetuses under the conditions of the present study. PMID- 11400314 TI - [Reproductive and developmental toxicity studies of S-1090, cefmatilen hydrochloride hydrate (4)--A study on oral administration during the perinatal and lactation periods in rats]. AB - Cefmatilen hydrochloride hydrate (S-1090) was administered daily by gavage to female rats at doses of 100, 300 or 1000 mg potency/kg/day from Day 17 of pregnancy to Day 20 of lactation to assess its effects on pregnant/lactating females and on development of the offspring. In dams, loose feces/reddish brown feces, increased cecum weight, decreased weights of the heart, spleen and submaxillary gland in all the S-1090 dosing groups and a decreased weight of the thymus in the 1000 mg potency/kg dosing group were observed. However, no effects on parturition and lactation were observed in any of the dosing groups. In F1 offspring, although increased cecum weight was found at weaning in all the S-1090 dosing groups, no abnormalities in viability, physical development, sensory functions/reflexes, behavior and reproductive function were observed. No adverse effects were observed in F2 fetuses and offspring. On the basis of these results, the no observed adverse effect levels of S-1090 are estimated to be less than 100 mg potency/kg/day for maternal general toxicity, and 1000 mg potency/kg/day for maternal reproductive toxicity and for developmental and reproductive toxicity in offspring under the conditions of the present study. PMID- 11400315 TI - [Immunological properties of S-1090, cefmatilen hydrochloride hydrate]. AB - S-1090, a cefmatilen hydrochloride hydrate, is being developed as a cephalosporin antibiotic for oral use. Immunogenicity, hypersensitivity-eliciting antigenicity and immunological cross-reactivity with other antibiotics were evaluated by active systemic anaphylaxis (ASA) test, passive cutaneous anaphylaxis (PCA) test and enzyme-linked immunosorbent assay (ELISA) using guinea pigs and mice/rats. In addition, in vitro direct Coombs' test was also performed to examine the possibility of hemolytic anemia in clinical use. Immunogenicity of S-1090 was not observed in guinea pigs after repeated immunization with S-1090 by ASA or PCA tests. Even in ELISA, only weak antibody production against S-1090 was found in some guinea pigs from the intraperitoneal groups showing the antibody titers only 10(1) to 10(2). When the sera collected from C3H/He mice and C57BL/6J mice immunized with S-1090 were tested for immunogenicity, rat PCA was elicited in a C3H/He mouse serum by S-1090 and antibodies against S-1090 were detected in a C57BL/6J mouse serum by ELISA. When adjuvant was used in mice and guinea pigs, the production of antibody against S-1090 was less frequent in comparison with other antibiotics such as cefmetazole (CMZ) and cefotiam (CTM). When hypersensitivity-eliciting antigenicity of S-1090 was examined using S-1090 as an eliciting antigen in ASA and PCA tests, positive ASA and PCA were observed in guinea pigs and positive PCA in a C3H/He mouse. Hypersensitivity-eliciting antigenicity was also observed in other reference antibiotics, i.e. cephalothin (CET), CMZ and CTM. Immunological cross-reactivity among S-1090, penicillin G (PCG), CET, CMZ and CTM was tested by ASA and PCA tests. S-1090 was found to immunologically cross-react only with CET in guinea pigs. In the present study, immunological cross-reactivities were also noted between PCG and CET, PCG and CMZ, PCG and CTM, and between CET and CMZ. In in vitro direct Coombs' test using human red blood cells, S-1090.Na, PCG and CET gave positive reactions at the final concentrations of 40 mg/mL, 20 to 40 mg/mL and 2.5 to 10 mg/mL, respectively. PMID- 11400316 TI - [Genotoxicity studies of cefmatilen hydrochloride hydrate (S-1090)]. AB - Cefmatilen hydrochloride hydrate (S-1090), a new non-ester type of orally active cephem antibiotic synthesized in Shionogi Research Laboratories, was evaluated for its genotoxic potential using three assay systems. In a reverse mutation test with bacteria of Salmonella typhimurium TA100, TA1535, TA98, and TA1537, and Escherichia coli WP2uvrA using the preincubation method, the number of revertant colonies in the S-1090 treated plates was almost equal to that in the negative control plates in all strains with and without metabolic activation system with S9 mix (maximum dose, 100 micrograms/plate in TA98). In a chromosomal aberration test with cultured Chinese hamster lung cells (CHL/IU), S-1090 did not induce structural chromosome aberrations or polyploid cells either in the absence or presence of S9 mix up to the 50% growth inhibition doses. The potential of inducing clastogenicity and/or disruption of mitotic apparatus in vivo by S-1090 was evaluated by a micronucleus test with bone marrow cells of male Jc1:ICR mice. S-1090 suspended in 0.5% aqueous methylcellulose was administered by oral gavage up to 2000 mg/kg/day in single and double dosing groups. No induction of micronucleated polychromatic erythrocytes was observed 24 hr after the last dosing in each group. As all three genotoxicity tests showed negative responses, S-1090 is thought to have no genotoxic potential. PMID- 11400317 TI - [Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (4)--One- and three month repeated oral dose toxicity studies in dogs]. AB - One- or three-month repeated oral dose toxicity studies of Cefmatilen hydrochloride hydrate (S-1090) were conducted in beagle dogs. Doses were set at 25, 100 and 400 mg potency/kg/day in both studies. In both studies, no deaths occurred, and reddish-brown feces (due to chelated products of S-1090 and its decomposition products with Fe3+ in the diet) were observed in all treated groups. A transient excretion of reddish urine was observed in the 400 mg potency/kg group and a slight increase in plasma irons was also observed in the 100 and 400 mg potency/kg groups of both studies. However, as no changes suggesting anemia or hepatic injury were noted in these groups, the change of plasma irons was considered to have no toxicological significance. Plasma S-1090 concentrations increased in a manner less than dose-proportional in both studies. In the one-month toxicity study, no toxicologically significant changes, including the above findings, were noted, so the NOAEL was assessed to be 400 mg potency/kg/day. In the three-month toxicity study, urinalysis in the 400 mg potency/kg group revealed a positive reaction to occult blood and erythrocytes in sediments. In the pathological examinations, submucosal edema, hemorrhage, inflammatory cell infiltration and occasionally focal mucosal thickening were observed in the urinary bladder of the 400 mg potency/kg group. The cystisis was considered to result from chronic stimulation with the metabolite(s) of S-1090 in urine, and the reversibility was demonstrable upon one-month drug withdrawal. From these results, the NOAEL of S-1090 in the three-month toxicity study was assessed to be 100 mg potency/kg/day. PMID- 11400318 TI - [Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (2)--Single oral dose toxicity study in dogs]. AB - Cefmatilen hydrochloride hydrate (S-1090) was administered at 500 and 1000 mg potency/kg once orally to beagle dogs. No deaths occurred. Vomiting, diarrhea or mucous feces occurred on the dosing day, and reddish-brown feces (due to chelated products of S-1090 and its decomposition products with Fe3+ in the diet) were also observed on the dosing and next day. Increases of plasma urea nitrogen and iron were observed on the next day after dosing. No remarkable changes were noted in other examination items. The animals in both groups were considered to be exposed to a similar level of S-1090 based on the toxicokinetic data. The oral lethal dose of S-1090 in dogs was estimated to be more than 1000 mg potency/kg. PMID- 11400319 TI - [Toxicity study of cefmatilen hydrochloride hydrate (S-1090) (5)--Six-month repeated oral dose toxicity study and supplement study in rats]. AB - Cefmatilen hydrochloride hydrate (S-1090) was orally administered to rats at dose levels of 100, 300 and 1000 mg potency/kg once daily for 6 months. All the S-1090 treated groups showed soft feces, reddish-brown feces (due to chelated products of S-1090 or its decomposition products with Fe3+ in the diet), abdominal distention, increased food and water consumption, lower urine pH, and a decrease of white blood cells counts (except for males of the 100 mg potency/kg group). One male in the 300 mg potency/kg group showed mucous feces and marked decrease in body weight, and diet in the middle stage of the administration period. In necropsy of the survivors of all treated groups, marked cecal enlargement was noted. No remarkable changes were observed in the other examination items. From the early stage of the withdrawal period, animals in the 1000 mg potency/kg group showed again soft or mucous feces and a marked decrease in body weight. Of these animals, one male died and another male was sacrificed in a moribund state at about 2 weeks of the withdrawal period. Enterocolitis was observed in these cases. Almost all animals recovered within 3 weeks of withdrawal. A supplemental study of the 6-month toxicity study was conducted to examine the mechanisms of enterocolitis and the changes observable in the 100 or 300 mg potency/kg groups after drug withdrawal. As a reference, cefdinir (CFDN), an oral cephem antibiotic the same as S-1090, was added in the 1000 mg potency/kg group. No deaths occurred in any groups. Decreased intestinal flora were noted in all the groups treated with S-1090 or CFDN at the end of the dosing period. At 2 weeks of the withdrawal period, C. difficile and its D-1 toxin in the cecal contents were highly detected in the S-1090 300 and 1000 mg potency/kg groups and CFDN group. Inflammatory changes in the cecum and colon were observed in these groups. At 4 weeks of the withdrawal period, intestinal flora in the S-1090 groups almost returned to the condition before dosing, but those in the CFDN group were retained highly. Cecal D-1 toxin in the CFDN group was positive and higher than in the S-1090 groups. It was thus considered that the critical condition with enterocolitis resulted from C. difficile, which proliferated more rapidly than the other bacteria and D-1 toxin produced by this bacteria in the withdrawal period. Above changes were commonly observed in the CFDN group. The NOAEL of S-1090 was assessed to be 100 mg potency/kg/day which induced no enteritis. PMID- 11400320 TI - [Does 'selective binding profile' result in 'selective treatment'?]. AB - It has been believed that the 'selective binding profile' may result in the 'selective treatment' for a disease. However, typical antipsychotics with the selective affinity for dopamine D2 receptor produce antipsychotic efficacy against positive symptoms accompanied with extrapyramidal side effects. Serotonin dopamine-antagonists produce fewer EPS, since they have a potent antagonistic activity for 5-HT2A receptor besides the dopamine D2 antagonism. The disinhibition of diminished glutamatergic transmission and alpha 1 antagonism by MARTAs (Multi-Acting-Acting-Receptor-Targeted-Antipsychotics) can reduce positive symptoms of schizophrenic patients, and the 5-HT2A antagonitic and anticholinergic activities of the drugs may reduce the EPS. Moreover, MARTAs increase intracellular concentrations of dopamine and norepinephrine in the prefrontal cortex and disinhibit the diminished glutamatergic transmission, which are expected to improve the negative symptoms, anxiety and depressive symptoms, and cognitive impairment that are difficult to be treated by other existing antipsychotics. These important biological changes are induced by the antagonism of 5-HT2A, 5-HT2C, alpha 1-adrenergic and muscarinic receptors by MARTAs. Thus a multireceptorial profile of MARTA is considered to result in the improved treatment. Therefore 'selective binding profile' does not always mean 'selective treatment.' Instead, if a breakthrough drug is to be developed for the treatment of refractory diseases, it may be necessary to rethink the ordinary strategy targeting for a single receptor. PMID- 11400321 TI - [Review of the research of glia maturation factor and cloning of human and rat glia maturation factor-gamma (GMFG) cDNA]. AB - Glia maturation factor-beta (GMFB) is a 17-kDa protein that was initially identified as a growth and differentiation factor acting on neurons as well as glia in the vertebrate brain. We isolated human and rat glia maturation factor gamma (GMFG) cDNA and examined the tissue distribution of GMFG in human and rat by Northern blots and Western blots. Sequence analysis of the entire cDNA revealed an open reading frame of 426 nucleotides with a deduced protein of 142 amino acid residues. The deduced amino acid sequence of its putative product is highly homologous to GMFB. Northern blot analysis indicated that a 0.9 kb mRNA is predominantly expressed in rat thymus, testis, and spleen. In comparison with GMFB, the current study demonstrated that the tissue distribution of GMFG is not the same as that of GMFB, and GMFG is predominantly in proliferative and differentiative organs. PMID- 11400322 TI - [Molecular pathogenesis of motor neuron diseases]. AB - Spinal and bulbar muscular atrophy (SBMA) and amyotrophic lateral sclerosis (ALS) are representative motor neuron diseases in which selective neuronal degeneration occurs. In this paper, some molecular aspects are discussed related to the pathogenesis of the neuronal degeneration. SBMA is a an X-linked neurodegenerative disease caused by the expansion of a CAG repeat in the first exon of the androgen receptor (AR) gene. To date, eight CAG repeat diseases have been identified, including spinal and bulbar muscular atrophy (SBMA), Huntington's disease (HD), dentatorubralpallidoluysian atrophy (DRPLA), and five spinocerebellar ataxias (SCAs 1, 2, 3, 6, 7). These disorders very likely share a common pathogenesis caused by the gain of a toxic function associated with the expanded polyglutamine tract. Several mechanisms have been postulated as a pathogenic process for neurodegeneration caused by the expanded polyglutamine tract. In SBMA, nuclear inclusions (NIs) containing mutant AR protein have been observed in regions of SBMA central nervous system susceptible to degenerations. Transcriptional factors or their cofactors, such as CREB or creb-binding protein (CBP) sequestrated in NIs, may alter the major intracellular transcriptional signal transduction and ultimately may result in neuronal degeneration. The components in the ubiquitin-proteasome pathway also colocalized in NIs and contribute to the path-ogenesis of SBMA. We generated two types of transgenic mice expressing 239Q under the control of human AR promoter and full-size AR containing 97Q. Marked neurological symptoms and extensive nuclear inclusions were observed in both transgenic lines, but there was no neuronal cell death, suggesting that major neurological phenotype was due to neuronal dysfunction instead of neuronal cell death. As for the therapeutic strategies, the overexpression of Hsp70 and Hsp40 chaperones acted together to protect a cultured neuronal cell model of SBMA from inclusion formation and cell death by mutant AR with expanded polyglutamine tract. In regard to ALS, we are screening the gene expression profiles of the motor neurons from the human ALS and SOD transgenic mouse spinal cord. Motor neurons were microdissected from the spinal cord samples by a lazer-captured microdissection system. Gene expression profiles were screened by cDNA microarray and molecular indexing. Several new molecules were cloned and characterized for their function and relation to neuronal cell dysfunction. Some molecules characterized in this procedure were briefly described. PMID- 11400323 TI - [Gene expression by addictive drugs and drug dependence]. AB - Drug dependence is developed by repeated administration of addictive drugs. This phenomenon resembles to learning and memory, although input stimuli are different. Pur alpha is abundant in the brain and binds to a single stranded DNA with (GGN)n sequence. The DNA-binding activity was enhanced by calmodulin and decreased in the brain treated with chronic morphine. The data suggest that morphine stimulates gene expression by a pathway different from that of cAMP. Arc protein is an "effector protein" and markedly induced by convulsion or methamphetamine. A novel protein, Amida, was isolated as an Arc-binding protein. Amida was transported into the nuclei and induced apoptosis and the inhibition of cell growth. The functions of these protein and the involvement of them in the development of drug dependence were discussed. PMID- 11400324 TI - [The involvement of phosphoinositide 3-kinase (PI3-Kinase) and phospholipase C gamma (PLC gamma) pathway in the morphine-induced supraspinal antinociception in the mouse]. AB - The present study was designed to investigate whether the phospholipase C gamma (PLC gamma)/phosphoinositide 3-kinase (PI3-Kinase) pathway could participate in the expression of the supraspinal antinociception induced by intracerebroventricular (i.c.v.) administration of mu-opioid receptor agonist in the mouse. The i.c.v. pretreatment with PI3-Kinase inhibitors, wortmannin and LY294002, and a specific antibody to PLC gamma 1 significantly attenuated the antinociception produced by either i.c.v. or systemic (s.c.) injection of a prototype of mu-agonist morphine. The s.c. injection of morphine produced a marked increase in the level of membrane-bound PLC gamma 1 isoform as compared to that from the saline-treated mice. This up-regulation of PLC gamma 1 by morphine was significantly inhibited by i.c.v. pretreatment with LY294002, indicating that morphine can activate PLC gamma 1 through the stimulation of PI3-Kinase. Pretreatment with a specific IP3 receptor inhibitor xestospongin C suppressed the morphine-induced antinociception in a dose-dependent manner. Recent studies have demonstrated that PI3-Kinase can be activated by G beta gamma, but not by G alpha subunit. In the present study, i.c.v. pretreatment with specific antibodies to G12 alpha and G beta gamma significantly suppressed the antinociception induced by morphine, whereas the specific antibody to Gq/11 alpha did not affect the antinociception induced by morphine. The present findings suggest that the supraspinal antinociception induced by mu-opioid receptor agonist may be mediated, at least in part, by the activation of PLC gamma through the stimulation of PI3-Kinase modulated by G beta gamma subunit. PMID- 11400325 TI - Better methods needed in detecting ADEs. AB - Rare incidents such as wrong-side surgery may grab the headlines, but the more common injuries--such as those caused by adverse drug events (ADEs)--are the ones that hospitals and health systems should be concerned about. PMID- 11400326 TI - Managing health care variability to achieve quality care. AB - While much has been written about variation and health care, one area that has received little attention is variation within hospitals related to the operations management--which can lead to wasted money and human resources. Two Boston researchers who have been studying this area say that addressing these variations -and using techniques found in other major industries across the country--could give hospitals a new tool in addressing patient safety issues, nursing shortages, cost containment, and overall better quality of care. PMID- 11400327 TI - Nurse staffing found to impact quality of care. AB - The size and mix of nurses in a hospitals throughout the country make a difference in the quality of care provided to patients, according to a Department of Health and Human Services study released last month. The study, Nurse Staffing and Patient Outcomes in Hospitals, found a consistent relationship between nurse staffing and outcomes for four medical conditions--urinary tract infections, pneumonia, shock, upper gastrointestinal bleeding--and length-of-stay. PMID- 11400328 TI - Ras regulation of urokinase-type plasminogen activator. PMID- 11400329 TI - Ras regulation of cyclin D1 promoter. PMID- 11400330 TI - Ras regulation of cyclin-dependent immunoprecipitation kinase assays. PMID- 11400331 TI - STAT proteins: signal tranducers and activators of transcription. PMID- 11400332 TI - Integrin regulation of receptor tyrosine kinase and G protein-coupled receptor signaling to mitogen-activated protein kinases. AB - Assays for use in integrin-mediated or -modulated signaling are essentially the same as those used in signaling studies involving growth factors or hormones. The major differences are the manipulations of the cells to compare the effects of gain or loss of anchorage, or the role of specific adhesion receptors. This needs to be done with some care, and with thought given to the overall biology of the particular cell type under investigation. Harsh treatment of cells, for example, prolonged suspension culture, may result in irreversible nonphysiological effects in some types of cells. PMID- 11400333 TI - Assaying activity of individual protein kinases in crude tissue or cellular extracts. PMID- 11400334 TI - R-Ras regulation of integrin function. PMID- 11400335 TI - Caveolin and Ras function. AB - Experimental protocols that allow confident assignment of signaling proteins to specific subdomains of the plasma membrane are essential for a full understanding of the complexities of signal transduction. This is especially relevant for Ras proteins, where the different membrane anchors of the Ras isoforms target them to functionally distinct microdomains that in turn allow quantitatively different signal outputs from otherwise highly homologous proteins. The methods outlined in this chapter, in addition to being invaluable in addressing Ras function, should also have wide utility in the study of many mammalian signal transduction pathways. PMID- 11400336 TI - Analyses of M-Ras/R-Ras3 signaling and biology. PMID- 11400337 TI - Analyses of TC21/R-Ras2 signaling and biological activity. PMID- 11400338 TI - Characterization of Rheb functions using yeast and mammalian systems. PMID- 11400339 TI - Ras regulation of skeletal muscle differentiation and gene expression. PMID- 11400340 TI - Induction of senescence by oncogenic Ras. PMID- 11400341 TI - Ras and Rho protein induction of motility and invasion in T47D breast adenocarcinoma cells. AB - There has been much interest in how Ras and Rho proteins affect cell migration and invasion. Studies of the molecular mechanisms and signaling pathways that are involved will be aided by the development and use of relevant model systems and simple in vitro assays. While a complete understanding of metastasis will ultimately need to employ in vivo studies, the migration and invasion assays presented here are good initial assessments of events that are relevant to the metastatic cascade. PMID- 11400342 TI - Ras regulation of vascular endothelial growth factor and angiogenesis. AB - Given the multifaceted role of Ras in tumor angiogenesis, pharmacologic targeting of such proteins may bring about at least three important consequences: (1) partial obliteration of the angiogenic competence of tumor cells, (2) an increase in vascular dependence and sensitization to apoptosis, and (3) a direct inhibition of endothelial cell responses to proangiogenic stimuli. Exploration of some of these possibilities, using various pharmacological compounds and antibodies, has already begun. An intriguing possibility is that Ras antagonists and signal transduction inhibitors may synergize with a number of other antiangiogenic modalities such as direct acting antiangiogenic agents (e.g., endostatin) or antivascular regimens involving low-dose continuous chemotherapy as a vasculature-targeting strategy. PMID- 11400343 TI - Recombinant adenoviral expression of dominant-negative Ras N17 blocking radiation induced activation of mitogen-activated protein kinase pathway. PMID- 11400344 TI - Ras regulation of radioresistance in cell culture. PMID- 11400345 TI - Paired human fibrosarcoma cell lines that possess or lack endogenous mutant N-ras alleles as experimental model for Ras signaling pathways. AB - We present here a human cell model for examination of mutant N-ras function. The HT1080 human fibrosarcoma cell line is pseudodiploid and contains a single endogenous mutant N-ras allele. MCH603c8 cells are a variant of HT1080 cells, in which the mutant allele has been deleted. The two cell lines differ dramatically in the constitutive levels of activation of downstream members of the Ras signaling pathways, and in biological features of transformation and tumorigenicity. Downregulation or activation of individual Ras-dependent pathways can be accomplished via transfection of dominant negatives or activated mutant cDNAs into HT1080 and MCH603c8 cells, respectively. The biochemical and biological consequences of expression of these mutant cDNAs can be assessed. There are dramatic effects on both the transformed and tumorigenic phenotype, depending on the cell line and mutant cDNA that is transfected. PMID- 11400346 TI - Determining involvement of Shc proteins in signaling pathways. PMID- 11400347 TI - Orally bioavailable farnesyltransferase inhibitors as anticancer agents in transgenic and xenograft models. AB - The in vivo evaluation process described here was instrumental in the identification of SCH 66336 as a clinical candidate. Our lead FTI, SCH 66336, and several other FTIs are being evaluated in early-phase clinical trials to establish proof-of-principle for farnesyl transferase inhibition in human patients. The preclinical studies described here suggest that FTIs may have utility against a wide array of human cancers as a single agent and may, at least in some cases, lead to tumor regression. In addition, the results to date in combination with cytotoxic chemotherapeutic agents in animal models indicate that these combinations may enhance the clinical efficacy of FPT inhibitors. Further preclinical studies should help to guide the clinical development of this class of novel antitumor agents. PMID- 11400348 TI - Animal models for Ras-induced metastasis. PMID- 11400349 TI - Nonradioactive determination of Ras-GTP levels using activated ras interaction assay. PMID- 11400350 TI - Measurement of GTP-bound Ras-like GTPases by activation-specific probes. PMID- 11400351 TI - Immunocytochemical assay for Ras activity. PMID- 11400352 TI - Ras activation of phosphatidylinositol 3-kinase and Akt. PMID- 11400353 TI - Assays for monitoring p70 S6 kinase and RSK activation. PMID- 11400354 TI - Ras activation of PAK protein kinases. PMID- 11400355 TI - Ras signaling to transcription activation: analysis with GAL4 fusion proteins. PMID- 11400356 TI - Ras regulation of NF-kappa B and apoptosis. PMID- 11400357 TI - Ras activation of NF-kappa B and superoxide. PMID- 11400358 TI - Ras, metastasis, and matrix metalloproteinase 9. PMID- 11400359 TI - Determination of quantity and quality of polyphenol antioxidants in foods and beverages. AB - The methods described in this article are quick, simple, and inexpensive to perform. The Folin quantitation method can determine both free and total polyphenol antioxidants in foods and beverages as described, as well as botanical extracts. This assay may also be used to estimate the daily per capita consumption of polyphenols in foods. The dose-response in vitro lower density lipoprotein antioxidant activity measurement (IC50) can be employed to compare antioxidants as pure compounds, or in mixtures after quantitating the polyphenols. The ex vivo lipoprotein-binding antioxidant activity can be measured simply and rapidly to determine possible in vivo binding of pure compounds or extracts from foods. Supplementation and epidemiology studies can utilize the rapid and inexpensive affinity column isolation method of lower density lipoproteins for the determination of lipoprotein oxidative susceptibility. PMID- 11400360 TI - Preparation and characterization of flavonoid metabolites present in biological samples. PMID- 11400361 TI - Caffeic acid as biomarker of red wine intake. PMID- 11400362 TI - Measurement of trans-resveratrol, (+)-catechin, and quercetin in rat and human blood and urine by gas chromatography with mass selective detection. PMID- 11400363 TI - Absorption of trans-resveratrol in rats. PMID- 11400364 TI - Analysis of complex mixtures of flavonoids and polyphenols by high-performance liquid chromatography electrochemical detection methods. PMID- 11400365 TI - Galvinoxyl method for standardizing electron and proton donation activity. PMID- 11400366 TI - Structure-activity relationships governing antioxidant capacities of plant polyphenols. PMID- 11400367 TI - Antioxidant and prooxidant abilities of foods and beverages. PMID- 11400368 TI - Metal chelation of polyphenols. PMID- 11400369 TI - Mechanism of antioxidant effect of catechins. PMID- 11400370 TI - Free radical scavenging by green tea polyphenols. PMID- 11400371 TI - Polyphenol protection of DNA against damage. PMID- 11400372 TI - Markers for low-density lipoprotein oxidation. PMID- 11400373 TI - Antioxidant activity of hydroxycinnamic acids on human low-density lipoprotein oxidation. PMID- 11400374 TI - Analysis of flavonoids in medicinal plants. PMID- 11400375 TI - Rapid screening method for relative antioxidant activities of flavonoids and phenolics. PMID- 11400376 TI - Nitric oxide formation in macrophages detected by spin trapping with iron dithiocarbamate complex: effect of purified flavonoids and plant extracts. PMID- 11400377 TI - Redox cycles of caffeic acid with alpha-tocopherol and ascorbate. PMID- 11400378 TI - DNA damage by nitrite and peroxynitrite: protection by dietary phenols. PMID- 11400379 TI - Overview of methods for analysis and identification of flavonoids. PMID- 11400380 TI - Repair of oxidized DNA by the flavonoid myricetin. PMID- 11400381 TI - Binding of flavonoids to plasma proteins. PMID- 11400382 TI - Protein binding of procyanidins: studies using polyacrylamide gel electrophoresis and French maritime pine bark extract. AB - The application of PAGE to determine the interaction between procyanidins and proteins, as presented here, enables one to directly determine the binding of either a pure of a complex mixture of flavonoids to a particular protein. If the protein of interest is an enzyme, the combination of PAGE with quantitative activity measurements allows identifying whether a change in the enzyme activity is related to the binding. Data presented suggest that PBE and EGb 761 have protein-binding properties, which, in addition to their redox-based effects, could provide a biochemical basis for their action in biological systems. PMID- 11400383 TI - Characterization of antioxidant effect of procyanidins. PMID- 11400384 TI - Inhibition of in vitro low-density lipoprotein oxidation by oligomeric procyanidins present in chocolate and cocoas. PMID- 11400385 TI - Biological actions of oligomeric procyanidins: proliferation of epithelial cells and hair follicle growth. PMID- 11400386 TI - Effect of polyphenolic flavonoid compounds on platelets. PMID- 11400387 TI - Assessing bioflavonoids as regulators of NF-kappa B activity and inflammatory gene expression in mammalian cells. PMID- 11400388 TI - Interaction between cultured endothelial cells and macrophages: in vitro model for studying flavonoids in redox-dependent gene expression. AB - This article focused on two methods to measure the activity of NF-kB. Both methods evalute "post-IkB phosphorylation" stages in the NF-kB activation cascade. In fact, EMSA performed with nuclear extracts provides an information only on NF-kB nuclear translocation and its ability to bind kB-DNA sequences. Likewise, the reporter gene assay is limited to assessing NF-kB-dependent gene expression no matter the mechanism that originally activated NF-kB. Nevertheless, the latter assay represents a more physiological and more reproducible way of measuring NF-kB activity in mammalian cells than the EMSA does. In order to obtain further insights into NF-kB signal transduction pathways, investigating IkB degradation and phosphorylation are recommended. The cloning and characterization of IkB kinases provided new testing possibilities based on measure of their activity. PMID- 11400389 TI - Determination of cholesterol-lowering potential of minor dietary components by measuring apolipoprotein B responses in HepG2 cells. PMID- 11400390 TI - High-performance liquid chromatography/mass spectrometry analysis of proanthocyanidins in food and beverages. PMID- 11400391 TI - Direct thiolysis on crude apple materials for high-performance liquid chromatography characterization and quantification of polyphenols in cider apple tissues and juices. PMID- 11400392 TI - Enzymes involved in hydroxycinnamate metabolism. PMID- 11400393 TI - Estimation of procyanidin chain length. PMID- 11400394 TI - Determination of flavonols in body fluids. PMID- 11400395 TI - [Report of the "Third International Conference on Priorities in Health Care". Will we really one day have to ration care?]. PMID- 11400396 TI - [Evidence-based medicine or patient-based medicine? And what if these two approaches end up not that far from one another...Or when the Oin-Oin syndrome is revisited!]. PMID- 11400397 TI - [Medical-economic analysis: a necessary complement to "evidence based medicine"]. AB - After an extraordinary technical development, medicine is under close scrutiny and requested to prove that its diagnostic and therapeutic procedures are "efficient, appropriate, and economical". Evidence based medicine allows the optimal use of existing data in the literature, but focuses only on health care benefits. Economical analyses take into account the health care resources needed to get these benefits. The various types of costs, perspectives of analysis, and techniques for assessing benefits and uncertainty about cost estimates, are presented and illustrated with two examples drawn from the fields of primary care and advanced technology. Because the cost benefit ratio of a diagnostic or therapeutic procedure is heavily dependent on the type of patients to which it is applied, as well as on the stage of technological progress, the two types of information are necessary to fully assess medical procedures. PMID- 11400398 TI - [Managed care, control of costs and the patient's health]. AB - The following article aims at reviewing the main historical evolutions of the Managed Care in the United States of America. We will also try to gain a better understanding of the main ethical issues raised by the physicians in the Managed Care context. Those issues have raised important concerns among the population of physicians. Furthermore, we will try to describe some of the mechanisms that have been developed by the HMOs to reduce the costs of care in the U.S.A. Finally, we will see the side-effects of the measures that have been decided to reduce the cost of care and how they affect the quality of care delivered to patients. PMID- 11400399 TI - [American hospitals in the managed care environment: strategy and perspectives]. AB - The following article aims at giving some informations about the solutions that have been launched by the american hospitals in order to improve their situation. These measures appear in the context of a tougher competition. But there are a lot of uncertainties on the future on the american hospitals. PMID- 11400400 TI - [Exposure in utero to immunosuppressives]. AB - The number of pregnant women receiving immunosuppressive therapy is increasing. Use of immunosuppressants during pregnancy is indicated for anti-rejection therapy in transplantation patients and treatment of autoimmune diseases. Despite the maternal and fetal risks of these pregnancies, the proportion of surviving infants is improving and the possibility that a pregnancy could occur in these women during their childbearing years should be considered. All immunosuppressant drugs and their metabolites cross the placenta, raising questions about the long term outcome of the children exposed to these agents in utera. There is no increased risk of congenital anomalies. However, there is an elevated incidence of prematurity, intrauterine growth retardation (IUGR) and therefore low birthweight, as well as maternal hypertension and preeclampsia. The most frequent neonatal complications are those associated with prematurity and IUGR, as well as adrenal insufficiency with corticosteroids, immunological disturbances with azathioprine and cyclosporin, and hyperkalemia with tacrolimus. The long-term follow-up of infants exposed to immunosuppressants in utero is still limited and experimental studies raise the question whether there could be an increased incidence at adult age of some pathologies including renal insufficiency, hypertension and diabetes. The follow-up of these infants should be carefully organized and multidisciplinary, taking the perinatal context into account. PMID- 11400401 TI - [Maternal phenylketonuria]. AB - The child of a phenylketonuric woman is exposed during pregnancy to a high risk of growth retardation and malformation. The frequency of these abnormalities is proportional to the maternal phenylalanine blood concentrations. If a strict low protein diet is followed before conception and throughout gestation the risks of abnormalities are not higher than in the normal population. The maternal blood phenylalanine levels must be maintained between 120 and 250 mumol/l and the tyrosine blood levels between 45 and 90 mumol/l. Weekly blood analyses are mandatory. Regular dietary controls are necessary to assure that the adequate energetic intake and the correct amounts of vitamins and minerals recommended for a pregnant woman are sustained. A case report is the opportunity to discuss certain practical aspects concerning the monitoring of the pregnancy of a phenylketonuric woman and to make general recommendations. PMID- 11400402 TI - [Goiters in children]. PMID- 11400403 TI - ["Urgent" pediatric consultations constantly increasing: a problem that affects all of French-speaking Switzerland]. PMID- 11400404 TI - [Methicillin resistant staphylococcus aureus (MRSA): incidence is growing in French-speaking Switzerland]. PMID- 11400405 TI - [Thrombopenias induced by heparin: diagnostic and therapeutic innovations]. AB - Heparin-induced thrombocytopenia (HIT) is a difficult immune-mediated phenomenon that physicians have to cope with. It is caused by platelet-activating, heparin dependent IgG antibody and may be associated with venous and arterial thrombosis, potentially fatal. Simple tests are now available which can detect most antibodies implicated in HIT. The old term "white clot syndrome" suggested that HIT was mostly associated with arterial thromboembolic events, but more recent publications indicate that HIT is also an important risk factor for venous thromboembolism. In case of suspicion of HIT, treatment by heparin should imperatively be stopped and an alternative anticoagulant treatment should be started before any laboratory confirmation. PMID- 11400406 TI - [Implantable defibrillator. Update]. AB - The implantable cardioverter-defibrillator is a device able to detect and efficiently treat life-threatening ventricular arrhythmias. Its decisive accomplishment in reducing sudden cardiac death and total cardiac mortality, opposed to the insufficient reliability of the traditional therapies explains its present ascendancy. In this review, the working principles and the implant techniques are developed, as well as the complications and the usual problems which could be encountered in implanted patients. Finally, the current indications are discussed in the light of recent clinical trials. PMID- 11400407 TI - [Did you say APLIM? Presenting the Association for Medical Information]. PMID- 11400408 TI - [Medicine based on proof (evidence based medicine)...or the Ouin-Ouin syndrome]. PMID- 11400409 TI - [ About Dr. Claude Miefille's article "who is insane?"]. PMID- 11400410 TI - [The last 10 years of Revista Espanola de Salud Publica in CD-ROM]. PMID- 11400411 TI - [The Goldthorpe Social Class Classification: reference framework for a proposal for the measurement of social class by the Working Group of the Spanish Society of Epidemiology]. AB - Two of the most important theory-based social class classifications are that of the neo-Weberian Goldthorpe and that of the neo-Marxist Wright. The social class classification proposal of the SES Working Group employed the Goldthorpe schema as a reference due to the empirical and mainly pragmatic aspects involved. In this article, these aspects are discussed and it is also discussed the problem of the validation of the measurements of social class and the problem of the use of the social class as an independent variable. PMID- 11400412 TI - [Cordylobia anthropophaga cutaneous myiasis]. AB - The progressive increase in the number of people traveling to tropical countries has led to imported diseases becoming of increasingly greater importance. Cutaneous myasis (or fly larva infestations) are found among this type of diseases particularly frequent in tropical countries. Based on the observation of a case of massive cutaneous myasis by Cordylobia antropophaga of a 34 year old woman upon her return from a trip to Senegal, a review has been made of the major cases of furunculoid cutaneous myasis published in Spain, as well as of the biology, pathology, treatment and prevention of human myasis by Cordylobia anthropophaga. The aforesaid case was characterized by the infestation of an unusually large number of larvae, the etiology of which was not suspected until the final stage of the disease. Although the cases of cutaneous myasis are not as serious as other imported diseases, a knowledge of this disease is necessary from the preventive, diagnostic and curative standpoint. It is important to proceed to the identification of the larvae by distinguishing them from another type of myasis involving different therapeutic implications. PMID- 11400413 TI - [Analysis of the mortality in a Catalonian rural population in the last third of the 19th. Century: Canet d'Adri, 1872-1900]. AB - BACKGROUND: We carried out a descriptive analysis of the mortality in a Catalonian rural town (Canet d'Adri, 12 km from Girona) for the period 1872-1900. The knowledge of the main mortality causes, especially those non-transmissible, could be very illustrative. The objective was to show the distribution of the mortality by age and cause, and to compare mortality due to infections with mortality consequence of the rest of the causes. METHOD: Three aspects were analysed: the distribution of the deaths by age and sex; the temporal and seasonal evolution; and the distribution by cause-specific. RESULTS: 49.3% of the deaths corresponded to women and 50.7% to men. Younger than one-year old were the most important group, 35.6% of the total mortality, followed by older than 65, 23.7%. Respiratory diseases were responsible for 33.4% of deaths; infectious diseases 31.1%, circulatory diseases 13.4% and digestive diseases 2.6%. Amongst younger than 45 year old the main cause of mortality was infectious diseases, above all in women. Respiratory diseases were the main cause of mortality in women older than 45 and in men older than 65 year old. Deaths because circulatory diseases were a very important cause amongst men older than 65. CONCLUSIONS: Besides of the importance of the infectious diseases as the main cause of death, it is surprising the importance of circulatory diseases (the third cause of mortality). PMID- 11400414 TI - [Case-control study of markers of oxidative stress and metabolism of blood iron in Parkinson's disease]. AB - BACKGROUND: Increasingly growing evidence exists of the involvement of oxidative stress mechanisms in Parkinson's disease. Lower levels of GSH in the sustantia nigra, an increase in iron buildup, an increase in the byproducts of lipid peroxidation and alterations in the mitochondrial complex I have been described. However, few studies have been made of levels of antioxidants in the peripheral bloodstream and of the influence of the intake of nutrients on the development of this disease. METHODS: In a group of 79 patients afflicted with idiopathic Parkinson's disease and a control group comprised of 107 subjects, compared by age, sex and place of residence, the lowered levels in the plasma of glutathione (GSH), malon dialdehyde (MDA), uric acid, tocophenol, beta-carotene, lycopene and different iron metabolism parameters were studied. Likewise, the intake of certain antioxidants was estimated based on a dietary survey. RESULTS: Significant differences (p < or = 0.001) were found in the plasma levels of GSH between cases (0.10 mumol/ml +/- 0.06) and controls (0.29 mumol/ml +/- 0.12). Likewise, the plasma levels of uric acid were lower (p < or = 0.05) in the cases (4.96 mg/ml +/- 1.96) than in the control groups (5.39 mg/ml +/- 1.13). No significant difference was found in the plasma levels of MDA, tocopherol, beta carotene and lycopene. With regard to iron metabolism, significantly higher ferritine and transferrin values were found in the patients with EP than in the control group, showing a lower transferrin saturation percentage (p < or = 0.05). The iron showed no significant changes between cases and control groups. CONCLUSIONS: The results of this study support the possible involvement of oxidative stress in the pathogenesis of Parkinson's disease and reveal, in turn, alterations in some peripheral blood parameters in keeping with known findings in the sustantia nigra. PMID- 11400415 TI - [Cytomegalovirus seroepidemiology in the community of Madrid]. AB - BACKGROUND: Cytomegalovirus (CMV) is frequently asymptomatic. However, it constitutes an important cause of congenital disease and severe pathology in immunodepressed patients, thus representing an important problem in Public Health. The object of this work was to study the prevalence of IgG against CMV (IgG-CMV) in the general population from Madrid. METHODS: It is a transversal study, in which IgG-CMV was assayed in a representative sample of the general population from the Region of Madrid, aged 2 to 60 years (n = 2030). Participants were recruited by a two-stage cluster sample procedure from those attending primary health care centres between October, 1993 and February, 1994. For the statistical analysis the chi 2 and chi 2 lineal trend tests were employed. The percentages of seroprevalence and the specific odds ratios were calculated with confidence intervals of 95%. RESULTS: The overall seroprevalence has been 62.8% (IC95% 60.6-64.9), ranging from 58.4% (IC95% 55.2-61.5) in men to 66.7% (IC95% 63.7-69.5) in women. A significant association between increase of the age and increment of the seroprevalence was observed. The consult to dentistry, the antecedent of surgery, as well as tattooing and acupuncture has been identified as risk factors of acquiring the infection. On the other hand, to have carried out University studies seems to act as a significant factor of protection. CONCLUSIONS: Although the risk factors detected indicates a transmission by blood, the high prevalence suggests the existence of other more common ways. The age-dependent seroprevalence increase confirms an important number of infections in the adult age. However, it cannot be exclude that this increase responds to an effect cohort due to socio-economic improvements similar to the detected for other virus. PMID- 11400416 TI - [Factors associated with reported vaccination coverage in early infancy: results of a telephone survey]. AB - BACKGROUND: Barcelona's Continuing Immunization Plan affords the possibility Of monitoring the immunization coverage of the population by means of the voluntary family postal notification system. Prior studies have revealed that some families fail to provide notification while being correctly vaccinated, which can lead to actual coverage being underestimated. The objectives of this study are to estimate the early childhood immunization coverage of the population and to ascertain the factors associated with failure to provide notification of immunization. METHODS: A phone survey was conducted on a sample of 500 children regarding whom there was no record of any notification of the first three childhood vaccine doses (diphtheria, tetanus, whooping cough and oral polio), in addition to a sample of 500 children who were on record as having been immunized. To estimate the actual immunization coverage, all children were considered to have been properly immunized when their family members did provide notification. As regards those who failed to reply, it was considered in the worst of cases that these were cases of children who had not be immunized. In the best of cases scenario, a coverage similar to those of the responses was assumed. RESULTS: The response to the questionnaire was higher among those who had previously provided notification of immunization by way of the postal notification system (79.1%) than among those who had failed to provide notification of immunization (67%). The leading factors associated with failure to report immunization status were the size of the families, the use of private health care services and the place of birth of the parents. Solely six (6) cases of those who had failed to report immunization admitted to not having immunized their children, totaling 1.9% of the responses. The immunization coverage of the population in question would total 99.7% in the best of cases and 93.7% in the worst of cases scenario. CONCLUSIONS: Immunization coverage of the population in question is quite high. The results underline the importance of promoting immunization notification among health care professionals, especially in the private sector. PMID- 11400417 TI - [Evolution of mortality attributable to tobacco in the Canary Islands (1975 1994)]. AB - BACKGROUND: The objective of this study is that of describing the death rate attributable to tobacco on the Canary Islands throughout the 1975-1994 period. METHOD: Deaths by age, sex and cause from 1975 to 1994 were obtained from the Spanish National Institute of Statistics (Natural Movement of Population). Based on the Spanish and Canary Island Health Surveys, the percentages of those who had never smoked, smokers and ex-smokers for the Canary Island population were taken by age and sex. The relative risks of death were taken from the Cancer Prevention Study II carried out in the United States. The percentages of deaths attributable to smoking were calculated for each year, sex and age group based on the attributable fraction of the population. Likewise, the trend in the death rate attributable for the time period in question was calculated and given in the form of the annual mean percentage change in the age-adjusted death rates by way of a log-linear model. RESULTS: During the 1975-1994 period, the number of deaths attributed to smoking rose by 64%. For major causes, a 108% increase in neoplasias, a 32% drop in cardiovascular diseases and a 15.5% increase in respiratory diseases were found for the period under study. The number of deaths was also found to increase with age, the 65 and over age group having been found to be that in which the most deaths caused by smoking occurred. CONCLUSIONS: On the Canary Islands, over 20% of all deaths in 1994 can be attributed to smoking. This suggests that the measures implemented to control the smoking habit are insufficient. PMID- 11400419 TI - [To know in order to transform. Information and knowledge for equity in health]. PMID- 11400418 TI - [Salmonella enteritidis outbreak in a home for the aged]. AB - BACKGROUND: An outbreak of gastroenteritis caused by Salmonella Enteritidis (SE), phagotype 1, reported at a senior citizen living facility in Burgos on November 15, 1999 is discussed. The objective of the research of this outbreak was that of ascertaining the source of infection, the mechanism by which it was transmitted and to propose the suitable measures for preventing any recurrence of this problem. METHODS: An observational retrospective cohort study has been made by surveying 106 of a total of 119 residents and 9 employees at the center in question. The Relative Risk of being affected by this disorder has been estimated in terms of the intake of different foods. Logic Regression was employed for calculating the odds ratio adjusted by age, sex and by the intake of foods and wine. The concordance of the clinical diagnosis with the coproculture diagnosis was studied using the Cohen's Kappa index of agreement. RESULTS: A total of 42 individuals reported symptoms (37 residents and 5 employees). The clinical attack rate was 36.5%. Forty-five (45) of the 91 coprocultures performed (82 on residents and 9 on employees) were positive (41 in residents and 4 in employees). The attack rate for cases confirmed by coproculture was 49.5% among those investigated. Fried breaded milk and flour batter sweets was the food product implicated epidemiologically with the onset of the outbreak: RR 3.25 (C.I at 95% 1.10-9.59). The protective effect of the wine (Odds Ratio: 0.68) was not high enough to be statistically significant (p = 0.36). The Kappa index was 0.43 (p < 0.001), which reveals very little agreement between the clinical diagnosis and the coproculture diagnosis. CONCLUSIONS: Epidemiological evidence exists of the intake of fired breaded milk and flour batter sweets having been the mechanism for the transmission of the infection, and the food product in question having been stored at room temperature having been the determining contributing factor in the start of the outbreak. This reveals that the best way of preventing this type of outbreaks are clean habits and practices on the part of all those involved in the food-handling sector. PMID- 11400420 TI - [The Habana declaration toward to equitable access to information in health]. PMID- 11400421 TI - [Analysis of the publicity campaign of the tobacco brand Fortuna in 1999-2000]. AB - The majority of tobacco advertisement in Spain is directed at youth. This advertisement, carried out on several fronts, is particularly effective with young people. In the present work we analyze different advertising efforts involved in the ad campaign carried out by Fortuna-brand (a product of the company Tabacalera S.A., and sector leader of sales in Spain) in 1999 and 2000. In terms of their marketing strategy, we can conclude that Fortuna's principal commercial objective is presenting tobacco as intrinsically associated with being young. Both the indirect promotion of its products through activities and its commercials, are aimed towards developing the image that tobacco symbolizes the series of core values more appreciated by youth. Relevant legislation will have to be adapted to this reality in order to control specifically the manner in which publicity such as that promoted by Fortuna is directed to the population. PMID- 11400422 TI - [Changes in socioeconomic differences in the utilization of and accessibility to health services in Spain between 1987 and 1995/97]. AB - BACKGROUND: The objective of this study is to evaluate the trend in the use and accessibility of health services in Spain between 1987 and 1995-1997 in groups with different socio-economic characteristics. METHODS: The data used are taken from National Health Surveys carried out on the adult population in 1987 and 1995 97 by the Ministry of Health and Consumer Affairs. The information for 1995 and 1997 has been combined due to the different sample size, in such a way that the estimates so obtained are the mean from both years. Population analysed has been people aged 24 years and older. The following have been studied: visits to the doctor, hospitalization, visits to the dentist and the gynecologist, the time taken to reach the visits to the doctor, and the waiting time in both the visits to the doctor and ordinary hospital admission. The socio-economic characteristics used are the level of education and the socio-economic group of the respondents. The measure of the association calculated between use of services and the socio economic characteristics was the percentage ratio estimated by binomial regression. In addition, the relative index of inequality was estimated as a global measure of inequality. RESULTS: Visits to the doctor were the most frequent factor in people without academic qualifications and the lower socio economic group in the two periods, while visits to the dentist and the gynecologist were more frequent in the higher socio-economic groups in both periods. Statistically significant differences in the frequency of hospitalization in the two periods were not found. Neither in 1987 nor 1995/1997 were statistically significant differences found between the different socio economic groups as regards the time taken to visits to the doctor (p > 0.05), although there were in waiting time in visits to the doctor (p < 0.05). In the second period the socio-economic differences observed in the first period in waiting time for ordinary hospital admission disappeared. CONCLUSIONS: In the second half of the 1990s the same socio-economic profile was observed in the use of health services and the time spent waiting to receive them, as in the second half of the 1980s, with the exception of waiting time for ordinary hospital admission in the second period. PMID- 11400423 TI - [Use of hypolipemic agents in Spain, 1987-2000]. AB - BACKGROUND: Lipid-reducing drugs are currently among those most used and which have the greatest impact on the drug spending of the Spanish National Health Care System. At the same time, constant changeovers are taking place from one type of molecule to another within this group. METHODS: A study was made of all of the Spanish National Health Care System prescriptions filled from the treatment subgroup B04A "Lipid-Reducing Preparations" nationwide throughout the 1987-2000 period. The different active ingredients have been grouped according to the subgroups of the Anatomical Therapeutic Chemical Classification. The measurement parameters employed were the number of packages, the economic cost, the number of Defined Daily Doses (DDD) and the Treatment/Day Cost (TDC). RESULTS: Lipid reducing drug consumption for the timeframe in question totals 389,133,314,142 pesetas (2,338,738,320 euros) and 2,580,179,790 DDD. Consumption rose annually, ranging from 4,504,321,869 pesetas in 1987 to 71,389,377,528 in 2000; and from 63,594,576 to 420,878,797 DDD. The TDC rose, in turn from 71 to 170 pesetas. The subgroup showing the highest consumption was that of the statin drugs, totalling 73.4% of the total consumption by cost and 54.8% of the number of DDD's, showing a trend toward further growth. CONCLUSIONS: Lipid-reducing drugs have undergone a noticeable, constant rise throughout the timeframe under analysis, not only regarding the price, but also the number of DDD's. Statin drugs having started to be used has modified the guidelines of use for this group. PMID- 11400424 TI - [Prevention secondary to ischemic cardiopathy at the lipid level in primary care in Aragon. PRECIAR study 1]. AB - BACKGROUND: Despite the existing evidence of the effectiveness of secondary prevention by modifying lifestyles or using the different drugs which have shown themselves to be clinically beneficial for heart patients, there is little, not highly accurate information available regarding the handling of the treatment of these patients within the scope of primary care in our country. The purpose of this study is that of ascertaining the current status of secondary prevention of heart disease as far as lipid control is concerned. METHODS: A review was made of the clinical records of all of the patients diagnosed at some point in time of their life as having ischemic heart disease, including those under the heading of acute myocardial infarction and angina pectoris within the 11 groups of patients assigned to three urban Health Care Centers in Area II in Zaragoza. The population in question totaled 19,692 patients, 388 cases of ischemic heart disease having been found in the record files data. RESULTS: Based on a study of the lipid control data, 60.8% of the cases analyzed in this study showed a complete lipid profile for the last year. Solely 4.7% of these patients had LDL < 100, the control objective having to be limited to c-LDL < 130 to find a 31.3% of patients with a lower than optimum control of this parameter. CONCLUSIONS: The results of our study reveal that the diagnosis of dyslipemias as well as the control of all other heart disease risk factors in secondary care have not been in keeping, for the most part, with the quality criteria of this health care procedure and therefore reveal the existence of a major degree of room for improvement. PMID- 11400425 TI - [Seroprevalence of three types of hepatotropic virus in a population of adolescents of the province of Guadalajara]. AB - BACKGROUND: Viral forms of hepatitis are one of the most infectious disease groups most often encountered in human pathology, and although most are benign, some can evolve into chronic forms. The purpose of this study is that of ascertaining the prevalence of hepatitis C, Epstein-Barr and human herpesvirus-6 infections among eighth-graders (13-14 years of age) and to related the same to variables regarding the gender and environment (rural or urban) in which they were living. METHODS: Descriptive, transversal study of a representative sample of the eighth-grade population in the province of Guadalajara during the 1998 1999 period. Systematic random sampling layered by gender and school (multi-stage sample format). The rate of specific IgG antibodies was determined using the ELISA technique. RESULTS: A study was conducted of 268 school-age children (46.6% males). The prevalence of Epstein-Barr antibodies was of 73.5% (CI: 67.9%-78.5%); 72% (CI: 64%-80%) among the males and of 74.8% (CI: 67.7%-81.9%) among the females (NS); 65.6% (CI: 57.4%-73.8%) residing in an urban environment and 80.7% in a rural environment (p < 0.05). The prevalence of those who tested positive for the hepatitis virus was that of 0.7%--two individuals, both females, one of whom resided in an urban environment and the other in a rural environment. For herpesvirus-6, the prevalence was of 82.4% (CI: 77.9%-86.9%); 81.5% (CI: 74.7% 88.3%) among the males and of 83.2% (CI: 77.1%-89.3%) among the females (NS), 86.7% (CI: 80.4%-92.2%) residing in an urban environment and 78.5% (CI: 73.1% 86.5%) in a rural environment (p < 0.05). CONCLUSIONS: The prevalence of Epstein Barr and human herpesvirus-6 and low degree as regards VHC is high, which tallies with the characteristics of the environment and age of the individuals under study. PMID- 11400426 TI - [Brucellosis as occupational disease: study of an outbreak of air-born transmission at a slaughter house]. AB - BACKGROUND: The onset of a large number of cases of brucellosis among employees at a slaughterhouse in Zaragoza where sheep from livestock culling procedures were being slaughtered, led to an investigation being made for the purpose of shedding light on the cause of this outbreak. METHODS: This study was organized into three parts: 1) Description of the outbreak 2) Study of the slaughterhouse layout and activity as regards the number of animals slaughtered, the employee workload and the degree of risk involved in each working area and 3) Unmatched case-control study. RESULTS: No significant differences were found among the attack rates by the sections of the slaughterhouse where the employees in question worked. The slaughtering analysis revealed a concomitant fluctuation between the slaughter of culled sheep and the epidemic. The case-control study revealed no significant differences for the risk factors involving working in a risk area, the use of protective measures and cuts/wounds. CONCLUSIONS: This outbreak is related to the slaughter of culled sheep. The fact that no significant difference were found by sections or due to working in a risk area is compatible with an overall explanation. Nor were any differences among the employees found to exist with regard to cuts/wounds or the use of protective measures, which focuses importance on airborne spread. PMID- 11400427 TI - Single-stranded regions in DNA isolated from different developmental stages of the sea urchin. AB - Long-term labeled sea urchin embryo (Strongylocentrotus purpuratus) DNAs were examined for size of recovered pieces, single-strandedness, and length of continuous double-stranded regions. Sizing on neutral sucrose gradients indicates that morula stage DNA sediments predominantly at 31 S, blastula stage DNA at 27 S, and gastrula stage DNA as a broad range of sizes of greater than 29 S. Treatment of [3H]thymidine-labeled DNA with Aspergillus oryzae S1 nuclease removes 19% of the 3H from morula stage DNA, 4% of the 3H from blastula stage DNA, and less than 0.1% of the 3H from gastrula stage DNA. Sedimentation of S1 nuclease treated [3H]DNAs on alkaline sucrose gradients indicates that in native morula stage DNA there is a nick or gap in one strand approximately every 9700 base pairs, in native blastula stage DNA about every 3300 base pairs, and very few nicks or gaps in native gastrula stage DNA. PMID- 11400428 TI - The effect of unilateral phrenicectomy on the rate of protein synthesis in rat diaphragm in vivo. AB - The fractional rate of protein synthesis (ks) in the denervated rat-diaphragm has been measured in vivo by the continuous amino acid infusion technique at 1, 3, 5 and 10 days after nerve section, and compared with the rate determined in normal rats. Similar rates of protein synthesis, 14% per day, were found for both the left and right hemidiaphragms in the control animals. In the denervated rats, the rates of protein synthesis in the contralateral control hemidiaphragms were significantly increased as soon as 1 day after nerve section. This is considered to be evidence of a compensatory synthesis in the control tissues. In the denervated hemidiaphragm, the rate of protein synthesis had doubled by the third day after nerve section, but by the fifth day had fallen slightly to a value some 50% greater than that of the controls, and remained at this level for a further 5 days. Based on these measured values of protein synthetic rate, calculated estimates have been made of the rate of protein degradation necessary to account for the reported (Turner, L.V. and Manchester, K.L. (1972) Biochem. J. 128, 789 801) changes in mass of the denervated tissue. During the first three days after nerve section, the rate constant for degradation increased to more than twice the normal rate for skeletal muscle, and remained at this value throughout the peak of the hypertrophy. PMID- 11400429 TI - Evidence against the incorporation into protein of amino acids directly from the membrane transport system in rat heart. AB - The possibility suggested recently [Hider, R.C., Fern, E.B. and London, D.R. (1969) Biochem. J. 114, 171-178; Hider, R.C., Fern, E.B. and London, D.R. (1971) Biochem. J. 121, 817-827; van Venrooij, W.J., Poort, C., Kramer, M.F. and Jansen, M.T. (1972) Eur. J. Biochem. 30, 427-433; and Adamson, L.F., Herington, A.C. and Bornstein, J. (1972) Biochim. Biophys. Acta 282, 352-365] that protein synthesis takes place using amino acids directly from the membrane transport system and not from an intracellular pool has been investigated in rat heart. The tissue was perfused first for 30 min with either [14C]glycine or [14C]leucine and then for a further 30 min with identical medium containing [3H]glycine or [3H]leucine, respectively. After an initial lag, [14C]glycine was incorporated into protein at a linear rate up to 60 min. The [3H]glycine was accumulated into tissue water and incorporated just as readily as the [14C]glycine had been. The rate of total protein synthesis agrees with literature values only if intracellular and not extracellular specific activity values are used in the calculation. Some glycine was converted to serine or threonine. Leucine influx and efflux were very rapid in contrast to the relatively slow exchange reported for incubated tissues [Hider, R.C., Fern, E.B. and London, D.R. (1969) Biochem. J. 114, 171-178; Hider, R.C., Fern, E.B. and London, D.R. (1971) Biochem. J. 121, 817-827; van Venrooij, W.J., Poort, C., Kramer, M.F. and Jansen, M.T. (1972) Eur. J. Biochem. 30, 427 433]. The results are consistent with the existence of an intracellular precursor pool for glycine. Some possible reasons for the discrepancies between this and the other studies are discussed. PMID- 11400430 TI - Escherichia coli initiation factor IF3 binding to AUG and AUG-containing single strands and hairpin loops, and nonspecific binding to polymers. AB - Nitrocellulose filter binding and equilibrium dialysis detected the binding of Escherichia coli initiation factor IF3 to AUG, An UGUm single strands and hairpin loops, poly(A,U,G), poly(U), and f2 RNA. No binding was detected for GUA, A8 U, or the hairpin loop A5 GC5 U5. AUG-specific binding, per nucleotide, is strong; nonspecific binding, per nucleotide, is weak. PMID- 11400431 TI - Isolation of a short, cytosine-rich repeating unit from the DNA of Escherichia coli. AB - Hybridization of heterologous nucleic acids has provided the means for isolating a repeating sequence which is located next to template regions of DNA. Separated single strands of 32P-labelled DNA from Escherichia coli were to a limited extent able to anneal with DNA of Micrococcus lysodeikticus immobilized on nitrocellulose membrane filters. The resulting hybrid was resistant to enzymes specific for unpaired strands, nuclease S1 (Aspergillus oryzae) and exonuclease I (E. coli). The E. coli DNA so hybridized was isolated and characterized. It contained all four bases with cytosine predominating; strand length was about 50 60 nucleotides. Since these units occupied about 1-2% of the length of the E. coli chromosome, they would have to be repeated about 2000 times in a single cell. Formation of the unusual hybrid was not diminished by prior saturation of the E. coli DNA with homologous 3H-labelled RNA. In fact both RNA and additional increments of DNA were detected on the filters approximately in a 1:1 ratio, showing that some of the repeating sequences were physically continuous with transcribed regions of DNA. PMID- 11400432 TI - Molecular structure of exonuclease I from Escherichia coli B. AB - Exonuclease I of Escherichia coli B (EC 3.1.4.25) was determined to be a monomeric protein of molecular weight approx. 72,000, as estimated by Sephadex gel filtration, sedimentation velocity centrifugation, and sodium dodecylsulfate polyacrylamide gel electrophoresis. PMID- 11400433 TI - The onset of collagen synthesis in sea urchin embryos. AB - In Arbacia punctulata and Strongylocentrotus purpuratus, two species of sea urchins, collagen synthesis begins during gastrulation and increases many-fold before reaching a plateau in the late pluteus stage. A collagen extraction method involving treatment with 0.1 M NaOH and hot 10% trichloroacetic acid provided the basis for a sensitive assay of collagen synthesis. PMID- 11400434 TI - The incorporation of wrong bases by DNA polymerase I following gamma-irradiation of DNA-like templates. AB - The synthesis of polydeoxyribose polymers by Escherichia coli DNA polymerase I has been investigated with control and gamma-irradiated DNA-like polymer templates containing only two bases. The results show that irradiation of a poly(dA) strand leads to the incorporation of dG, whereas irradiation of poly(dC) and poly(dG) strands both lead to the incorporation of dA. Irradiation of poly(dT) does not lead to the incorporation of any wrong base. The wrong bases are incorporated into the complementary strand of the newly synthesised DNA. PMID- 11400435 TI - Poly(A)-associated RNA in plants. AB - The RNA associated with poly(A) sequences from Euglena gracilis and Vicia faba has been isolated by binding to millipore filters and characterized by sedimentation velocity centrifugation and electrophoretic mobility. Poly(A) associated RNA as isolated in solution was highly aggregated. When denatured, it sedimented as a broad peak with a mean value of 16-18 S. This RNA was shown to be covalently linked to poly(A) sequences which are 150-250 nucleotides long. Our size estimates for plant poly(A) and poly(A)-associated RNA are similar to those obtained for animal cells. PMID- 11400436 TI - Differential effects of analogs of cycloheximide on protein and RNA synthesis in Achlya. AB - Analogs of the glutarimide antibiotic cycloheximide were tested for their effect on growth and incorporation of proline and uridine into acid-insoluble material in Achlya bisexualis. Each of the compounds tested had reduced antibiotic activity as compared to cycloheximide. The effects of the antibiotics on protein and RNA synthesis were varied. While cycloheximide inhibited both protein and RNA synthesis immediately, two of the analogs inhibited proline incorporation without effect on uridine incorporation, while three, each representing a modification of the hydroxyl of cycloheximide, stimulated uridine incorporation and either had no effect on or inhibited protein synthesis. These results indicate that the control of RNA synthesis by protein synthesis in Achlya can be released by glutarimide antibiotics. PMID- 11400437 TI - Gel electrophoretic analysis of poly(riboadenylic acid). AB - It is shown that molecular weights and molecular-weight distributions of poly(rA), and by implication other single-stranded polynucleotides, and synthetic and natural polyelectrolytes in general, can be determined by electrophoresis in polyacrylamide gels. It is shown that fractions of very narrow molecular-weight distribution can be obtained by preparative electrophoresis of polydisperse samples. Molecular-weight calibrations based on sedimentation coefficients of such fractions are given, and in aqueous systems do not coincide with calibrations for partially base-paired RNA species. Poly(rU) fractions fall on the same calibration as poly(rA). Relations between mobilities, relative to standard markers, and molecular weight for poly(rA) over a wide range of molecular weights are given, which allow rapid molecular-weight determination on poly(rA) samples, such as the segments found in many types of messenger RNA. PMID- 11400438 TI - Inhibition of uridine transport in cultured mammalian cells by theophylline. AB - Theophylline (theobromine, caffeine) reversibly inhibits the incorporation of labeled RNA precursors both in confluent 37 RC and in exponentially growing HeLa cells. As measured in 37 RC after 2 h labeling, 20 mM theophylline reduces the incorporation of [3H]UTP and [14C]uridine into acid-precipitable material to 5% and 9% of the control, respectively. This reduction is paralleled by a comparably lowered incorporation of the same precursors into the acid-soluble pool. The initial rate of incorporation into total cell material is similarly affected by theophylline, the inhibition being of a simple competitive type. Theophylline does not alter the turnover rate of pulse labeled RNA during actinomycin D chase nor does it preclude the utilization of the endogenous pool of nucleoside phosphates. Upto a concentration of 10 mM, it does not inhibit uridine kinase neither in 37 RC nor in HeLa cells. The mentioned inhibitory effects of theophylline cannot be mimicked by exogenously added cyclic AMP. All the data support the conclusion that theophylline inhibits the transport of uridine into the cell. PMID- 11400439 TI - The isolation from ribonucleic acid of substituted uridines containing alpha aminobutyrate moieties derived from methionine. AB - The RNA of an established line of Chinese hamster cells growing in cell culture contains a small number of uridines substituted at the 3-position with a gamma linked alpha-aminobutyrate residue. Structure has been ascertained by: (a) examination of incorporation of isotopic labels from precursors; (b) degradation with anhydrous hydrazine and comparison of the products with synthetic material or with hydrazinolysis products of known uridines; and (c) comparison of the unknowns as their hydantoin derivatives with the 5-beta-(bromoethyl)hydantoin alkylation products of uridine and of 1- and 3-methylpseudouridine. In this manner it is shown that 18-S RNA of ribosomes contains a single residue of a nucleoside which we tentatively identify as 1-methyl-3-gamma-(alpha-amino-alpha carboxypropyl)pseudouridine per molecule. RNA isolated from the supernatant fraction, sedimenting at 4 S and co-electrophoresing with transfer RNA on polyacrylamide gels, contains several similar bases, one of which is identified as 3-gamma-(alpha-amino-alpha-carboxypropyl)uridine. Each of the above nucleosides derives its alpha-aminobutyrate residue from methionine. PMID- 11400440 TI - The role of aminoacyl-tRNA binding site on the factor EF-T in uncoupled GTPase reaction. AB - The properties of the N-tosyl-L-phenylalanyl chloromethane treated EF-T factor were studied in a ribosomal system in which splitting of GTP occurs. The action of N-tosyl-L-phenylalanyl chloromethane inhibits the binding of aminoacyl-tRNA to the EF-T factor. The binding and exchange of guanosine phosphates continued to be preserved. The inhibited factor is inactive in the GTPase reaction which depends on the participation of ribosomes and aminoacyl-tRNA. The uncoupled GTPase reaction (which is not dependent on the presence of aminoacyl-tRNA) is also sensitive to the effect of the inhibitor. The inhibition of the uncoupled GTPase is incomplete. These findings are attributed to the involvement of the aminoacyl tRNA binding site of the EF-T factor in the uncoupled GTPase reaction. PMID- 11400441 TI - Immunologic studies on nucleic acids and their components. II. Reversible inhibition of anti-nucleoside antibodies in aqueous pyridine and its application in antibody purification. AB - The effect of aqueous pyridine on a hapten-antihapten system was investigated by the quantitative precipitin reaction and by the membrane filtration method. It was found that dilute solutions of pyridine inhibited the reaction between isopentenyladenosine and its antiserum. Other solvents examined were less effective. The effect of pyridine was reversible at concentrations where complete inhibition occurred, thus indicating its use for the dissociation of antigen antibody complexes. The inhibitory effect of pyridine was exploited in a single step purification method for anti-isopentenyladenosine and anti-deoxy-adenylate antibodies. In addition, generally applicable methods for linking nucleosides and nucleotides to aminoethyl-Sepharose are described. PMID- 11400442 TI - Informational ribonucleoprotein particles of newt oocytes: polyribosome associated ribonucleoproteins. AB - Various species of rapidly labelled, informational ribonucleoproteins can be isolated from homogenates of newt oocytes. Polyribosome-associated ribonucleoprotein can be separated from heterogeneous nuclear ribonucleoprotein and free cytoplasmic ribonucleoprotein by sucrose gradient centrifugation. The polyribosome-associated ribonucleoprotein can be released from the ribosome complex by treatment with low concentrations of EDTA and has the following properties: 1. It is rapidly labelled with [3H]uridine under condition (incubation of oocytes for 4 h and less at 20 degrees C) where there is no detectable labelling of ribosomal subunits. 2. It is heterogeneous in size, consisting of particles most of which sediment between 40 S and 80 S. 3. Its sedimentation coefficient is related directly to the size of the polyribosomal complex from which it is derived. 4. Its density ranges from 1.35 g/cm3 to 1.55 g/cm3 irrespective of size. This indicates protein to RNA ratios of 4:1 to 2:1. 5. It is active, when complexed with ribosomes, in cell-free protein synthesis. It is concluded that this polyribosome-associated ribonucleoprotein is functional messenger and its role in oocyte maturation is discussed. PMID- 11400443 TI - [Use of low-frequency ultrasound with special gas agents in acute generalized peritonitis]. AB - Optimal parameters of peritoneal cavity treatment by low-frequency ultrasound with alternative use of carbon dioxide and oxygen were selected in experimental animals. Sanitation of peritoneal cavity in fecal peritonitis led to a significant desintoxication effect. The results of treatment of acute generalized peritonitis by this method in 28 patients were good. On day 2 or 3 after surgery all the patients underwent extracorporeal detoxication with hemosorbent AUSRITC-1 with good result. In severe toxemia and water-electrolyte disturbances plasmapheresis was applied. 4 patients of 28 died, lethality was 14.2%. PMID- 11400444 TI - [Laparoscopic cholecystectomy in elderly and old patients]. AB - The results of cholecystectomy from mini-approach (CEMA) in 111 elderly and old patients with acute and chronic cholecystitis living in European North of Russia were analyzed, and also 84 patients were operated by traditional approach (TCE). Duration of CEMA was less than TCE (75 +/- 3.2 and 95.2 +/- 4.6 min respectively; p < 0.05). Sutures after CEMA were removed on day 8.4 +/- 1.2 (after TCE--on day 13.8 +/- 2.4, p < 0.05). Postoperative period after CEMA was 11.4 +/- 2.1 days vs. 18.8 +/- 3.5 days after TCE (p < 0.05). Complications after CEMA were seen in 1.8% patients, after TCE--in 5.0%. Lethality was 0.9% after CEMA and 3.5% after TCE. The main advantages of CEMA are: reduction of surgery time, early activation of patients, decrease of postoperative complications number and reduction of postoperative treatment time (11.4 +/- 2.1 days after CEMA and 18.8 +/- 3.5 days after TCE, p < 0.05). PMID- 11400445 TI - [Cancer of the gall bladder]. AB - The experience of treatment of 65 patients with cancer of the gall bladder (CGB) is analyzed. The tumor was revealed in women 7.2 times more often than in men. 95.3% patients were older than 60 years. Diagnosis of CBG is difficult before and during operation. Retrograde cholangiopancreatography increases reliability of diagnosis. The survival after radical cholecystectomy was 23.3 months. PMID- 11400446 TI - [Differential diagnosis of hemorrhagic fever with renal syndrome and acute surgical diseases of abdominal organs]. AB - The results of differential diagnosis of hemorrhage fever with renal syndrome and acute surgical diseases of abdominal organs in 1250 patients were analyzed. It was revealed that the course of the disease was similar to acute abdomen in 13.2% of them. The combination of fibrogastroduodenoscopy and diagnostic laparoscopy is optimal for differential diagnosis and permits to reveal typical symptoms of hemorrhage fever with renal syndrome. PMID- 11400447 TI - [Combined local use of miliacil and laser in the combined therapy of trophic ulcer]. AB - The efficacy of local use of a new drug miliacil in combination with laser radiation was studied in 44 patients with trophic ulcer of different origin. It is established by clinical, histological and microbiological examinations that healing of trophic ulcers is better in combined use of miliacil and laser radiation. Mean hospital stay reduced 1.3 times after this treatment. Miliacil and laser radiation were applied for preparation of trophic ulcer for autodermoplasty. Mean time of preparation of trophic ulcer for final closure was 1.5 times less than in isolated use of miliacil. Mean hospital stay of patients who underwent autodermoplasty was 1.3 times shorter. PMID- 11400448 TI - [Treatment of pyonecrotic complications in diabetic macroangiopathy]. AB - Results of treatment of 104 patients aged from 47 to 82 years with stage IV talocrural ischemia (by A.V. Pokrovsky) due to diabetic macroangiopathy were analyzed. Significant depression of immune status was revealed: number of T- and B-lymphocytes decreased by 40-50%, phagocytosis--by 65-75%, number of immunoglobulins--by 25-30%. The majority of the patients had endotoxicosis, hypercoagulation and hyperlipidemia; the correction of it by hemosorption and plasmapheresis was performed depending on the disease stage. 90 patients underwent surgery. Reconstruction in aorto-ileo-femoral zone was performed in 57 patients (in 34--with good results), in femoro-popliteo-femoral zone--in 33 patients (in 19--with good results). The "Gore-Tex" grafts were the plastic material for reconstruction, but autovein in situ was also used in femoro popliteal zone. PMID- 11400449 TI - [Surgery of a purulent wound of the foot]. AB - The experience of plastic closure of purulent wounds in the foot of 55 patients is presented. The average area of wounds was 61 cm2. Treatment of defects of plantar surface was most difficult. The differential policy in plastic surgery of foot wounds was developed. Complete closure of the defect was achieved in 81% patients. The most promising method of plastic closure is dosed tissue expanding. PMID- 11400450 TI - [Laparoscopic rectum repair]. AB - A reliable and simple method of allograft fixation in laparoscopic posterior-loop rectopexy (LPR) was developed. The study was carried out on 40 cadavers, peak effort of separation of the net fixed to the sacrum by various methods was evaluated. Effort of separation in fixation of the net by sutures was 10.1 +/- 2.12 kg, in fixation by hernial stapler--1.13 +/- 0.36 kg, stapler "Pro-Tack"- 6.38 +/- 1.85 kg, in fixation of the net by the new device--8.9 +/- 0.75 kg. From 1995 till 1999 LPR was performed with the new device in 15 patients, the open posterior-loop rectopexy (OPR) with fixation of the net by non-absorbable sutures -in 16 patients. In LPR intraoperative blood flow was twice as small (146.0 +/- 79.2 ml) as in OPR (275.6 +/- 76.9 ml). Mean time of LPR was 183.1 +/- 69.8 min, OPR--211.3 +/- 57.9 min. Relief of pain after LPR enabled with reduction of narcotic analgetics requirement (58.3 +/- 1.5 mg). Mean dose of narcotic analgetics after OPR was 93.2 +/- 1.4 mg. The proposed method of allograft fixation and device for it realization are effective and meet all requirements. PMID- 11400451 TI - [Surgical treatment of gastroduodenal ulcer hemorrhage]. AB - The results of treatment of patients with gastroduodenal ulcerous bleeding over 20-year period were analyzed. From 1997 active individualized policy was used in the treatment of 522 patients. In duodenal ulcer the drug-thermal vagotomy was applied as well as traditional organ-saving operations, in combination with extraduodenisation of ulcer when it penetrates into pancreatic head. In gastric ulcer sparing resection or methods of ulcer's taking outside gastrointestinal tract (extragastration of ulcer) with preservation of gastric angio-, and neuroarchitectonics were preferable. In preoperative period Sandostatin and Hystodil were applied, in postoperative period--Imunofan. PMID- 11400452 TI - [Errors and complications in the treatment of hemorrhoids]. AB - Long-term results of 724 hemorrhoidectomies were followed up from 6 months till 7 years. Various complications after operation were revealed in 172 (23.3%) patients. In 46 (26.7%) patients the failures were associated with unrecognized proctological diseases before operation, in 51 (30%)--with removal of hemorrhoids and in 63 (36.6%) patients the concomitant diseases of other organs and systems were the causes of complications. PMID- 11400453 TI - [Advanced techniques in surgical treatment of penetrating gunshot wounds of the chest]. AB - The results of treatment of 322 wounded with penetrating gun-shot wounds of the chest during local hostility from 1994 till 1996 are analyzed. Aggravation in the condition of the hospitalized wounded was observed due to reduction of time of evacuation in hospitals. The indications for surgery were in 96.3% wounded. The main surgical methods were thoracocentesis and drainage of pleural cavity performed in 80.1% wounded. Thoracotomy was performed in 9.6% wounded. High efficiency of operative videothoracoscopy performed in 4.4% wounded in hospital of the first aid is demonstrated, it allowed to reduce frequency of thoracotomy to 2.4%. PMID- 11400454 TI - [A plastic surgery method in vast defects of the anterior abdominal wall]. AB - The developed method of plastic repair of anterior abdominal wall's vast defects is intended for use in reconstructive-plastic herniology for closure in layers of total defects of the abdominal wall without reduction of abdominal cavities volume, in oncology and traumatology--for repair of abdominal wall's integrity after excision of massive tumors or damaged tissues. In correction by this method of large muscular-aponeurotic defects of abdominal wall the own tissues of the patient (greater omentum, hernial sac) and lavsan explants of author's construction are used maximally. New abdominal white line is formed, all the loted or insufficient tissues of abdominal wall are replaced. 88 patients with defects of anterior abdominal wall of different origin were operated by this method. Long-term results were followed up for 11 years, there was no recurrence, in 3 patients moderate relaxation in the area of hernioplasty was revealed, due to nonobservance of recommendations in early postoperative period. Patients with large hernias and defects of anterior abdominal wall should be operated in specialized surgical clinics. PMID- 11400455 TI - [Long-term observation of Crohn's disease]. PMID- 11400456 TI - [Ten simultaneous operations in one patient]. AB - In a patient, a woman of 62 years, one surgical team has performed consecutively over 3 h 45 min 10 operations for different diseases: extirpation of the uterus with appendages, removal of retroperitoneal fibrosarcoma, appendectomy, 30 cm resection of the small intestine, polypectomy from the rectum, creation of sygmorectoanastomosis, plastic reconstruction of postoperative ventral hernia with duplication, sectoral resection of mammary gland, removal of papilloma and lipoma of abdominal wall. The outcome is favorable. PMID- 11400457 TI - [Neuroendocrine tumors of the thymus]. PMID- 11400458 TI - [History of surgical treatment of gastric and duodenal ulcers in Russia (end of 19th - 20th century)]. PMID- 11400459 TI - [Acute pancreatitis after resection of stomach for low duodenal ulcer]. AB - Results of the treatment of 71 patients with postbulbar ulcer of the duodenum are presented. Stomach resection by Bilrot-II was performed in 22 (30.9%) patients, by Bilrot-I--in 2 (2.8%), by Roux with plastic reconstruction of duodenal stump with intestinal tube--in 16 (22.5%) patients. Precardial proximal vagotomy with drainage operation was performed in 31 (43.6%) patients with massive periulcerous infiltration. Acute postoperative pancreatitis (APP) was seen in 29 (40.8%) patients chiefly after resection of stomach, 6 (8.4%) of them had pancreonecrosis. Postoperative lethality in the group of patients with APP was 12.6%. Ultrasonic examination of the pancreas and its vascular structures is the main method for diagnosis of postoperative pancreatitis. The cause of APP is the mechanic deformation and spasm of major duodenal papilla (MDP) that may be associated with cholinergic denervation and edema of MDP after resection and trauma of tissues. The removal of MDP spasm is an effective prophylactic measure for elimination of pancreatic hypertension. One of the methods of removal of MDP spasm is periarterial sympathectomy of gastroduodenal artery and prolonged drug blockade of celiac plexus in addition to stomach resection. PMID- 11400460 TI - A decision support process to compare riparian revegetation options in Scheu Creek catchment in north Queensland. AB - While riparin vegetation can play a major role in protecting land, water and natural habitat in catchments, there are high costs associated with tree planting and establishment and in diverting land from cropping. The distribution of costs and benefits of riparian revegetation creates conflicts in the objectives of various stakeholder groups. Multicriteria analysis provides an appropriate tool to evaluate alternative riparian revegetation options, and to accommodate the conflicting views of various stakeholder groups. This paper discusses an application of multicriteria analysis in an evaluation of riparian revegetation policy options for Scheu Creek, a small sub-catchment in the Johnstone River catchment in north Queensland, Australia. Clear differences are found in the rankings of revegetation options for different stakeholder groups with respect to environmental, social and economic impacts. Implementation of a revegetation option will involve considerable cost for landholders for the benefits of society. Queensland legislation does not provide a means to require farmers to implement riparian revegetation, hence the need for subsidies, tax incentives and moral suasion. PMID- 11400461 TI - The environmental impact of water markets: an Australian case-study. AB - Water markets are developing as part of a Council of Australian Governments initiative to promote an efficient use of Australia's water resources. The consequences of these policies on river health is yet to be fully understood, but recognised as having significant interrelationships which need to be explored. This paper examines the consequences of introducing trade and allocating water for environmental use in the Border Rivers region of Queensland. The results of this study suggest that: (1) trade in water entitlements is likely to increase the differential between extractive demand and historical flow regimes as extractive water-use concentrates on the most profitable crops, and (2) water markets are likely to limit the effectiveness of water policies aimed at restoring natural flow regimes. As a result, trade-offs between environmental needs and income from extractive use will need to be determined. This work is important and timely in water-policy development demonstrating the linkages and trade-offs between ecological and economic objectives. PMID- 11400462 TI - Prediction of the impact of logging activities on forest cover: a case-study in the east province of Cameroon. AB - The objective of this study was to test the value of the concept of the net commercial value of standing timber in predicting the impact of logging activities on forest-cover modifications. A study area was selected in the East province of Cameroon which contains major primary forests and which contributes strongly to national timber production. A Geographic Information System containing ecological and economic variables was used in combination with remote sensing data to define the net commercial value of standing timber in the East province. Taking account of the potential commercial value of standing timber improves our understanding of the spatial determinants of logging activities and of the resulting forest-cover modifications. The occurrence of logging-induced forest-cover modifications increases with the value of forest rent. In one of the study sites, half of the very high rent areas have already been logged. In that site, therefore, it is mostly the low rent or marginal forest areas that remain unlogged. This was not the case, however, throughout the study area as shown by the observations at another site. PMID- 11400463 TI - A comparison of waste-reduction practices and new environmental paradigm of rural and urban Chinese citizens. AB - This paper compares the public attitudes of three southern Chinese cities/towns, namely Guangzhou, Dongguan, and Yuanzhou on waste recycling and avoidance, and the New Environmental Paradigm (NEP). The latter describes the concept of treating the resources and the ecosystems on Earth as things having their own rights rather than being subordinate and subservient to humans. These three places are chosen to represent areas in different stages of urbanization in the Guangdong province. As Hong Kong is also a southern Chinese city, where data of similar nature are available, comparison with Hong Kong is also made. It was found that the support for source separation of household waste in the rural and urban areas in mainland China was greater than that found in Hong Kong. Concerning actual practices on waste separation, about 85% of the respondents in the cities and almost all of the rural household surveyed have the habit of saving up and selling the recyclables. It is estimated that about 11-13% of the household waste was diverted in such a way. This is higher than the estimated 8% household waste recovery rate of Hong Kong. The majority of the respondents in the three surveyed cities have recovered household recyclables in the past but with higher family incomes, it was now less common. On the contrary, the lower income group tended to recover greater portion of their waste for selling to the waste depots. These findings have implications on the future waste management policy formulation in China. Regarding the acceptance of the New Environmental Paradigm, it was found that the rural population scored better than the urban population and they tended to agree more frequently with the NEP. However, the rural population perceives a greater potential for industrial and urban development without dampening environmental quality. It was also found that the NEP scores of the mainland Chinese are higher, in general, than their Hong Kong counterparts as measured in early 1990s. Insufficient public support has often been a concern for source separation programmes in all countries despite the obvious waste management benefits of recycling. This study shows that there is overwhelming support for source separation of waste in Mainland China, whether it is in rural or urban communities, and that most people are already undertaking source separation due to the redemption value of the recyclables. Therefore, for policy-makers, the concern should not be on whether the public knows how to separate the waste but rather on how to motivate them to separate waste for the good of society, and how to maintain an effective market for recyclables. PMID- 11400464 TI - Exploring red deer culling strategies using a population-specific calibrated management model. AB - Wildlife management is generally carried out under conditions of uncertainty. The exact population size is unknown, its future dynamics are uncertain and clear management objectives are often not formulated. In order to provide management advice in this situation, a framework is presented for combining different sources of information using a Bayesian approach for calibrating a management model. Harvesting strategies can then be explored based on predictions of future populations size and structure which incorporate parameter uncertainty. This method makes it possible to evaluate the probability of achieving certain objectives with different management strategies. The advantage of the approach presented in this paper lies in that both the model and the harvesting strategies are adaptable to any particular population of interest. The approach is illustrated for two Scottish red deer populations for which culling strategies corresponding to different management objectives are explored and their benefits evaluated. It is found that each population requires different culling rates for keeping population number stable, demonstrating the benefits of the population specific calibration of the management model. PMID- 11400465 TI - Developing decision support systems for integrated coastal management in the tropics: is the ICM decision-making environment too complex for the development of a useable and useful DSS? AB - Integrated coastal management in the tropics requires the conservation of vulnerable and diverse ecosystems such as coral reefs and mangroves as well as the management of land and marine-based human activities. Decision-making for integrated coastal management involves multiple decision-makers and multiple stakeholders often with conflicting needs and interests. Decision support systems can be developed to improve our understanding of the inter-relationships between the natural and socio-economic variables and hence result in improved decision making. The question is whether this decision making environment is actually too complex for the development of useful and useable decision support systems. This paper describes the components of the decision making environment and the components of a decision support system. It also explores the various techniques available to deal with different modelling needs, the constraints of inadequate data and the multi-objective decision making environment. In addition, different techniques of developing decision support systems can play important roles within integrated coastal management. Three coastal decision support systems are evaluated in terms of their design and role in integrated coastal management and are used to evaluate the potential to develop decision support systems for integrated coastal management. PMID- 11400466 TI - Economic valuation of urban forest benefits in Finland. AB - Urban development projects may cause loss of amenity values of green areas, which should be taken into consideration in planning. Therefore, quantitative information on residents' valuation concerning urban forests is needed for assessing urban land use. The purpose of this investigation was to study the valuation of urban forests in two different urban environments Joensuu and Salo, Finland. The aims were to study the attitudes towards and benefits related to the use of urban forests and, in particular, to measure the valuations in monetary terms using contingent valuation, i.e. measure the residents' willingness-to-pay for larger wooded recreation areas and for small forested parks. Urban forests were seen in both towns as clearly producing positive benefits rather than causing negative effects. The negative features of forests were related to the management of the areas rather than their existence. The main values were related to nature and social functions of forests. In contrast, timber production achieved a distinctively low priority in both study towns. The results stress the importance of defining urban forest policies for municipalities in Finland. More than two-thirds of the respondents were willing to pay for the use of recreation areas. Good location and active management raised the average WTP. Moreover, approximately half of the respondents were willing to pay for preventing construction in urban forests. The results also show that the monetary value of amenity benefits in recreation areas is much higher than the present maintenance costs. The examples concerning the advantageousness of construction on green areas suggest that a limit could be found where the infill of housing areas is not worthwhile from the point of view of society, if the losses of green space benefits are taken into account. PMID- 11400467 TI - Application of the Australian river bioassessment system (AUSRIVAS) in the Brantas River, East Java, Indonesia. AB - Assessment of river 'health' using biological methods, particularly those based on macro-invertebrates, is now commonplace in most developed countries. However, this is not the case in most developing countries, where physical and chemical methods are used to assess water quality, with very little use of biological assessment methods. This paper reports on a project that aimed to assess the possible introduction of biological assessment of river condition using the Australian River Assessment System (AUSRIVAS) into Indonesia. The paper addresses three components of the project: (1) science--does the bioassessment method work in this tropical region? (2) resources--are they adequate and if not what additional resources are needed? (3) politics--what needs to be done to convince the agencies (both central and provincial) to take up such a new philosophy and approach? A pilot study was run in the upper Brantas River, East Java. A total of 66 reference sites and 15 test sites were sampled and the macro-invertebrates collected were identified to family level. A rigorous quality-control protocol was introduced to ensure the data were reliable and reproducible. The macro invertebrate data were used to develop a predictive model of the AUSRIVAS type for the upper Brantas River, and the model was then used to assess the 'health' of sites that were presumed to be damaged in this section of the river. A number of difficulties were experienced during the study, including: locating reference sites sufficiently unmodified by humans; lack of skills to identify animals collected; and a paucity of facilities required for aquatic macro-invertebrate identification (e.g. identification keys and good quality binocular microscopes). For resources, the major constraint to the introduction of a bioassessment capability in Indonesia is the lack of personnel trained in the bioassessment techniques. An 'on-the-job' training approach was adopted, largely because of the specialist nature of this work. Six Indonesians were trained and will now become the 'trainers' of further Indonesian scientists (we have called this process 'training-the-trainers'). For politics, it was hoped that the AUSRIVAS method would be suitable for introduction into the Indonesian national Clean River Program. A strategy was developed and implemented to ensure the method and its outputs were accepted technically by the Indonesian scientific community, and also by the resource managers and relevant government officials. Experience shows that if the latter do not see how the bioassessment information will be used for management purposes they will not accept the method even if it is scientifically sound. PMID- 11400468 TI - Ascorbic acid loss in vegetables: adequacy of a hospital cook-chill system. AB - This research evaluates the ascorbic acid loss of vegetables prepared in a hospital cook-chill plated system. The ascorbic acid loss of three vegetable samples (carrots, peas and potatoes) were measured at various points in the cook chill system and analysed using the DCPIP method. Substantial losses for all vegetables were identified at the cooking and regeneration stages. The ascorbic acid levels of the selected vegetables when raw ranged from 6 to 10 mg/100 g sample. At the end of the food service cycle the ascorbic acid content of the vegetables was reduced to 1.7-5.8 mg/100 g sample. The largest percentage loss of ascorbic acid, i.e. 76%, was found in mashed potatoes. Peas incurred the least percentage loss, i.e. 42%. This progressive deterioration during the cook-chill plated catering system causes an insubstantial presence of ascorbic acid in the food served to hospitalised patients. PMID- 11400469 TI - Effects of energy density and sweetness of gruels on Burkinabe infant energy intakes in free living conditions. AB - In free living conditions, 24 breastfed infants, aged 6 to 10 months, were given successively five experimental gruels to study the effect of energy density (ED) and sweetness (sweet taste) on energy intakes (EI). Four gruels (G0, G1, G9 and G20) were prepared with experimental flours which were composed of the same local ingredients and which contained different levels of sucrose. The fifth gruel (GC) was prepared with an industrial flour. G0 had an average ED of 45 kcal/100 g (189 kJ/100 g) and the other gruels an average ED of 110 kcal/100 g (461 kJ/100 g). Although the sugar contents of the flours were 1% for G1, 9% for G0 and G9 and 20% for G20, because of flour composition and gruel dry matter content, the gruel G1 had the same sweetness as G0, G20 the same sweetness as GC and G9 a sweetness between that of G1 and G20. The results show that the amounts of G0 consumed were significantly higher than those of high ED gruels (7.84 for G0 vs 6.12, 5.63, 4.46, 4.72 g/kg body weight/meal, respectively for G20, G9, G1 and GC, P < 0.05). However, EI from high ED gruels were significantly (P < 0.001) higher than those from G0 (6.65, 6.10, 4.86, 4.83 kcal/kg/meal, respectively for G20, G9, G1 and GC vs 3.46 for G0). Energy intakes from G9 and G20 gruels were not significantly different but were significantly higher than those from GC and G1 (P < 0.001). So, consumption of sweet gruels with high ED and composed of local ingredients increased, at least by 76%, the EI from gruels in comparison with those from low ED gruels, but the amounts consumed by the infants remained too low to cover more than 15% of their daily total energy needs. PMID- 11400470 TI - Nutritive value of protein from sea urchin, and its effects on intestinal leucine aminopeptidase and intestinal and hepatic gamma-glutamyltranspeptidase. AB - Seafoods are a rich source of easily digestible protein that also provides polyunsaturated fatty acids, vitamins and minerals for human nutrition. The purpose of this study was to determine the nutritive value of diets high in protein from two species of sea urchin (Paracentrotus lividus and Echinus esculentus) as compared to a high-quality reference protein such as casein, as well as the effects of these diets on leucine aminopeptidase and gamma glutamyltranspeptidase, intestinal and hepatic enzymes. The test was carried out on three groups of male rats fed these diets for 23 days. The result of the nutritive parameters indicated that sea urchin, Echinus esculentus, were significantly lower than the casein protein except for food intake, and the nutritive parameters of sea urchin, Paracentrotus lividus, were similar to the values obtained for casein protein except for digestibility and net protein utilization. Intestinal leucine aminopeptidase activity was significantly lower (P < 0.05) in the group fed Paracentrotus lividus as compared to the group fed casein. Intestinal gamma-glutamyltranspeptidase showed a significantly higher activity in the group fed E. esculentus than in the two other groups and the hepatic gamma-glutamyltranspeptidase activity in the groups fed the two species of sea urchin were lower. Nutritive values suggested that these marine species are a good protein source and similar to casein. The activity of the enzymes under study must have been affected by the amino acid composition of the protein. PMID- 11400471 TI - Growth deceleration and bone metabolism in nutritional dwarfing rats. AB - Nutritional status as well as energy and protein intake are critical regulators of IGF-1 and IGFBP-3 and contribute to the modulation of bone remodeling and formation. The purpose of this study was to investigate on an experimental model with nutritional dwarfing (ND), whether the alterations on body growth velocity, energy metabolism and body composition could affect serum concentrations of IGF-1 and IGFBP-3 and bone (tibiae and mandible) histology and histomorphometry. Twenty one male weanling Wistar rats (body weight = 38.20 +/- 0.94 g) were randomized to three groups: seven of them were killed at day = 0 (CO, n = 7); control (C, n = 7); and experimental 80 (E80, n = 7). During 4 weeks, C was fed ad libitum with a 1:1 carbohydrate to fat diet. E80 was being underfed with the same diet by 80% and the following parameters were measured: weight (Wt) for length (L) ratio z score; oxygen consumption (VO2); body composition (BC) by EM-SCAN SA 3000. At t = 28, E80 and C were killed. Serum IGF-1 and IGFBP-3 and bone histology and histomorphometry were performed on C0, E80 and C. E80 showed Wt for L z-score between lean and adequate, a decrease in VO2 according to body proportions, a BC of a delayed puberty individual, IGF-1 and IGFBP-3 decreased by 56 and 53%, respectively. Tibiae's hematopoyetic and adipose bone marrow areas were combined, with sealing trabeculae on metaphyseal areas. This study suggests that there is a relationship among growth deceleration in ND rats and structural alterations on tibiae. PMID- 11400472 TI - A simple bacterial turbidometric method for detection of S. aureus. AB - A simple and rapid turbidometric method based on specificity of lysosotaphin to lyse S. aureus exclusively has been developed for detection of S. aureus contamination in foods. PMID- 11400473 TI - Mineral content of commercial pollen. AB - Pollen is a natural product which is extending its marketing day by day, given that it is considered to be a dietetic product and it is consumed everyday by a broad sector of the Spanish population. In its composition it presents valuable nutrients, among which we can find minerals, which is the main object of this study. We have analysed sodium, potassium, magnesium, copper, iron, manganese and zinc in 15 pollen samples which correspond to different brands. The technique we have used is atomic absortion spectroscopy. The results show us the great potassium contribution of this natural product, with values over 400 mg/100 g, and about microelements, mainly iron and zinc, although with different results, depending on the brand which markets it, with average values of 4.01 +/- 1.00 and 3.66 +/- 1.02, respectively. PMID- 11400474 TI - Characterization and fat migration of palm kernel stearin as affected by addition of desiccated coconut used as base filling centre in dark chocolate. AB - The characterization and fat migration of palm kernel stearin (PKS) and desiccated coconut, used as base filling centre in dark chocolate were studied. C36 and C38 triglycerides of PKS decreased by 11% and 9.6% respectively, whereas C32 and C34 increased by 97% and 48% respectively. The change in the triglycerides composition of PKS shift the melting point of PKS from 33.2 to 31.4 degrees C. Solid fat content (SFC) of PK reduced by 40% at 30 degrees C. The rate of fat migration was very slow at 18 degrees C storage compared to 30 degrees C. The rate of change of C36 in the chocolate layer was 0.1% week-1 and 1.2% week-1 at 18 and 30 degrees C respectively. Chocolate stored at 18 degrees C showed post hardening during storage period and withstood bloom during the storage period, whereas that stored at 30 degrees C became soft and bloomed faster after 3 weeks of storage. PMID- 11400475 TI - Characterization of vitamin B12 compounds from edible shellfish, clam, oyster, and mussel. AB - Vitamin B12 content of various edible shellfish was determined by both Lactobacillus leichmannii ATCC 7830 microbiological and intrinsic factor chemiluminescence methods. The values determined by the microbiological method were 1.2-19.8 (M/C ratio) fold greater in the shellfish than the values determined by the chemiluminescence method. Vitamin B12 compounds were purified from most eaten shellfish, oyster (M/C, 1.5), mussel (M/C, 1.2), and short-necked clam (M/C, 2.7), and partially characterized. TLC and HPLC patterns of each red colored vitamin B12 compound (M/C, 1.0-1.2) purified from these shellfish were identical to those of authentic vitamin B12. Although the higher values in the determination of vitamin B12 by the microbiological method may be due to the occurrence of vitamin B12-substitutive compounds, the edible shellfish would be excellent vitamin B12 sources judging from the values (> or = 6 micrograms/100 g) determined by the chemiluminescence method. PMID- 11400476 TI - Assessment of total energy expenditure in a Chinese population by a physical activity questionnaire: examination of validity. AB - A physical activity questionnaire from which total daily energy expenditure (TEE) could be estimated was developed for adult Hong Kong Chinese subjects, and its reliability and validity examined. The questionnaire was based on questionnaires used in Caucasians, and adapted for local lifestyle after focus group meetings involving subjects of all age groups. The questionnaire was administered to 94 subjects, consisting of healthy adults, the elderly, and two patient groups (those with renal disease on continuous ambulatory peritoneal dialysis and those with cancer). Seventy-one subjects were reinterviewed within 14 days to test reliability. Validity was examined in 31 normal subjects by measuring the basal metabolic rate (BMR) by indirect calorimetry and multiplying by the physical activity level (PAL) obtained from published studies using the doubly labelled water method and also from FAO/WHO/UNU to obtain the TEE. The intraclass correlation coefficient of reliability rages from 0.7 to 0.8 for all subject groups. The mean estimated TEE from the questionnaire was not significantly different from the mean value derived from measured BMR x PAL. The mean bias ranged from an underestimation of 27 kcal to overestimation of 215 kcal. However, the limits of variability were wide. Age was inversely related to the energy expended for occupational activities, but was positively associated with energy expended in leisure activities. Women spent less energy on occupational and exercise activities, and more on caretaking activities. Those with disease were also less likely to participate in caretaking activities. We conclude that this questionnaire may be a useful tool for future studies where energy expenditure needs to be estimated in various settings in the Hong Kong Chinese population. PMID- 11400477 TI - Fatty acids and lipids of camel milk and colostrum. AB - Total fat content was 32.8 +/- 14 g/L in camel milk (10-240 days post partum) or 30.1 +/- 19.5 g/L in colostrum (1-7 days post partum). Triacylglycerols accounted for 96% of the total lipids in milk. Triacyglycerols of camel milk contained saturated fatty acids (66.1%) and unsaturated fatty acids (30.5%). The predominant saturated fatty acids were 16:0 (34.9%) 14:0 (14.5%) and 18:0 (9.7%). The content of these fatty in acids in colostrum was lower (52.4%) than that of mature milk. The colsotrum contained a relatively high amount of 18:1 (25.4%), and 16:1 (13.9%), with the remainder being a mixture of dienoic and trienoic long chain fatty acids. Triacylglycerols contained low amount of short-chain fatty acids (C4-C8). There is a high content of polyunsaturated fatty acids in camel colostrum and milk. PMID- 11400478 TI - Spectrophotometric determination of the tannin contents of various Turkish black tea, beer and wine samples. AB - This study reports tannin contents of various tea (19), beer (6) and wine (6) samples, produced or sold in Turkey under different brand names. Determinations were carried out by employing a previously reported UV-vis spectrophotometric method. The standard addition procedure was also compared to the direct determination procedure and found to be more reliable. The tannin contents of hot water extracts of tea, tea bag and herbal tea samples ranged between 6.20-8.33, 8.03-6.59 and 2.76-6.54 percent (w/w) respectively. Tannin contents of beer and wine samples determined without extraction were found ranging between 66.36-77.26 and 67.18-107.62 micrograms/mL respectively. Iron contents of analysed wine samples were also found ranging between 0.36 and 10.33 ppm by AAS and no relation is found between tannin and iron contents. PMID- 11400479 TI - [Usefulness of p53 in the clinical management of bladder tumors]. AB - The antioncogene p53 has been the subject of many studies in the field of bladder cancer, for many years and there appears to be a good correlation between p53 protein overexpression and pejorative clinicopathological factors, but contradictory results have been reported concerning its prognostic value. These discordances can be partly explained by the various immunohistochemical methods used in published series. Standardization of a highly sensitive and specific immunohistochemical method for the detection of p53 alterations now appears to be essential in order to accurately determine the value of p53 in the clinical management of bladder cancer. PMID- 11400480 TI - [Chromophobe cell renal cancer, a distinct entity. Report of a series of 35 cases]. AB - OBJECTIVE: To describe the clinical and histological characteristics and natural history of clear cell renal carcinoma. PATIENTS AND METHODS: The case files of 35 patients (22 men and 13 women, with a mean age of 58 +/- 13.8 years) operated for clear cell renal carcinoma were reviewed. The circumstances of diagnosis and the histological features of the tumour were analysed. The outcome of the patients was studied with a mean follow-up of 70 +/- 20.2 months. RESULTS: The diagnosis was incidental in 66% of cases. The macroscopic appearance of the tumour was characteristic: homogeneous and beige or white colour. On light microscopy, tumours were composed of variable proportions of clear cells and eosinophilic cells. The were classified as 15 predominantly eosinophilic tumours (43%), 9 predominantly clear cell tumours (26%) and 11 tumours composed of equal number of clear cells and eosinophilic cells (31%). Nuclei were irregular and indented. Hale stain was positive in every case. The majority of tumours were confined to the kidney (T1: 65% and T2: 29%) with a low nuclear grade (91% of grades 1 and 2). No patient developed local recurrence or metastasis during follow-up. The overall 5-year survival rate was 92%. No death related to the disease was reported. CONCLUSION: The histological diagnosis of clear cell renal cancer is now easy and can be suggested even on macroscopic examination of the operative specimen. This tumour appears to have a good prognosis, as it is usually confined to the kidney with a low nuclear grade. PMID- 11400481 TI - [Role of helical tomodensitometry in the early diagnosis of renal infarction]. AB - INTRODUCTION: Renal infarction is a rare and often difficult diagnosis. The objective of this study was to demonstrate that contrast-enhanced spiral CT in patients presenting features of renal colic, can establish the diagnosis by confirming the presence of infarction of the renal parenchyma. MATERIAL AND METHODS: Over a 10-month period, the authors proposed the following decision flow chart for all patients admitted with clinical features of renal colic: plain abdominal x-ray and first-line renal ultrasound, which, in the absence of a diagnosis (stones or dilatation of cavities), were completed by unenhanced spiral CT scan. When these examinations were normal, contrast-enhanced spiral CT scan was then performed. RESULTS: 300 patients were included in this study, and CT was performed in 40 cases. This management allowed the diagnosis of infarction of the renal parenchyma in three patients, who are described here. When unenhanced CT sections do not reveal any abnormality, contrast-enhanced sections are essential to visualize the infarcted zone, seen as a triangular low density lesion with clearly defined margins and a vascular topography. CONCLUSION: Contrast-enhanced spiral CT should now be considered to be the reference examination for the assessment of non-documented renal colic. This recent imaging modality should allow the diagnosis of a greater number of infarctions of the renal parenchyma, which formerly remained undiagnosed. PMID- 11400482 TI - [Retrospective comparative study of extended nephrectomies by surgery and by retroperitoneal laparoscopy]. AB - OBJECTIVE: To evaluate the results of radical retroperitoneal laparoscopic nephrectomy in terms of tolerance, morbidity and oncologic control. MATERIAL AND METHODS: 22 radical nephrectomies have been carried out by retroperitoneal laparoscopy. One surgical conversion has been necessary. This series has been retrospectively compared to 16 radical nephrectomies by open surgery. RESULTS: Both series were comparable. Operative time was not significantly different in the two groups. Blood loss was less with laparoscopic surgery (p < 10-3). We did not see any difference in antalgic consumption during the first post-operative day and in intensive care and hospital stay. Post-operative complications were less in the laparoscopy group (p = 0.04). We observed no death or reccurrence with a mean follow-up of 8.7 months for the laparoscopy group vs 32.2 for the open group. CONCLUSIONS: Radical retroperitoneal laparoscopic nephrectomy is a recent surgical technique which has the same oncologic standards as open surgery. Blood loss and post-operative complications are significantly reduced. Survival needs to be evaluated on long term follow-up. PMID- 11400483 TI - [Natural history of vascular ports for hemodialysis after renal transplantation]. AB - INTRODUCTION: The objective of this study was to assess the long-term outcome of AV shunts in renal transplant recipients, to discuss mechanisms affecting their functioning and the surgical strategy designed to optimally preserve the venous capital in the hypothesis of a return to dialysis. MATERIALS AND METHODS: 160 renal transplant recipients, with a mean age of 47 years, were reviewed. AV shunts were performed at the wrist in 95% of cases and in the cubital fossa in 13% of cases. The AV shunt had been performed an average of 29 months before renal transplantation. RESULTS: 62% of AV shunts were considered to be functional with a mean follow-up of 69 months after transplantation and 95 months after creation. The intraoperative and early and late postoperative thrombosis rates were 6%, 7.5% and 17%, respectively. The AV shunt was subsequently closed in 12 patients (7.5%). CONCLUSION: Native distal AV shunts, although not used after renal transplantation, have a prolonged survival. The main risk is thrombosis which can be prevented intraoperative and perioperatively. These results encourage a conservative attitude to all well tolerated AV shunts. PMID- 11400484 TI - [Arterial embolization in hemorrhagic renal angiomyolipoma]. AB - OBJECTIVE: Renal angiomyolipoma (AML) is a rare benign tumour (2-3%) comprising variable proportions of an adipose component, which is often the most abundant, a smooth muscle contingent and a vascular contingent. The objective of this study is to demonstrate the value of embolization of bleeding angiomyolipomas. MATERIAL AND METHODS: 2 patient, aged 22 and 62 years, presented with perirenal haematoma secondary to bleeding angiomyolipoma and were treated by selective arterial embolization. RESULTS: In one case, partial nephrectomy was subsequently performed because of the size of the angiomyolipoma (10 cm). In the other case, attempted partial nephrectomy at 3 months failed due to the proximity of the renal hilum and nephrectomy was performed. CONCLUSION: Beyond a diameter of 4 cm, the risk of perirenal haemorrhage is proportional to the size of the tumour. When surgery is required, selective arterial embolization is the first stage of treatment allowing partial nephrectomy in the majority of cases. PMID- 11400485 TI - [Interstitial cystitis: reevaluation of patients who do no respond to standard treatments]. AB - OBJECTIVE: Insterstitital cystitis (IC) is an uncommon disease without an existing positive diagnostic test. Our experience has been that it tends to be overly considered, with many patients treated for long periods with several different regimens and no improvement. This impression prompted a review of our recent experience in the exclusionary diagnosis of IC. MATERIAL AND METHODS: A retrospective review of patients referred to our specialty clinic between December 1995 and October 1998 revealed 23 patients (21 women, 2 men) who had been diagnosed with IC, and had received at least one treatment for this disease (intravesical DMSO or Heparin, Elmiron, 1-Arginine, or therapeutic hydrodistension) with little or no benefit. Our own evaluation of these patients included urinalysis, non-invasive uroflow, post void residual determination and office cystoscopy (all patients), filling cystometrogram and pressure-flow-EMG studies (21 patients), voiding cystourethrogram (17 patients), urethral MRI (9 patients), and other studies as indicated. The results of our reevaluation are reported. RESULTS: Of the 23 patients referred with a diagnosis of IC, only 4 were considered to meet the diagnostic criteria as established by the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) after a thorough evaluation. One other patient did not meet the criteria but responded to medications for IC. Urethral pathology was a common finding (distal periurethral fibrosis causing obstruction in 5 patients, intraurethral wall diverticulum in 2, and chronic urethritis in 2), particularly among women with abnormal pressure flow studies during urodynamics. CONCLUSIONS: In our experience, many patients who carried the diagnosis of interstitial cystitis were found to have other causes contributing to their urinary symptoms after careful reassessment. Urodynamic studies were found to be of paramount importance in excluding the diagnosis in many cases. On the basis of our findings, we recommend that if local urethral tenderness is noted on physical examination or an abnormal pressure-flow relationship is seen during urodynamics, further investigation is warranted. PMID- 11400486 TI - [Reduction of sensitivity at the level of the membranous urethra after substitutive orthotopic ileal bladder]. AB - OBJECTIVE: To determine whether the sensitivity of the membranous urethra is an important factor to obtain continence after ileal bladder replacement. MATERIAL AND METHOD: The sensitivity threshold after electrical stimulation of the membranous urethra by a ring electrode placed on a urethral catheter was measured in 47 men after ileal bladder replacement (postoperative group). The control group consisted of 35 men before radical prostatectomy or cystoprostatectomy. RESULTS: The mean sensitivity threshold of the membranous urethra was 9 +/- 2 mA in the control group and 27 +/- 11 mA in the postoperative group (p < 0.001). The sensitivity threshold of daytime continent and incontinent patients was 24 +/- 9 and 39 +/- 10 mA, respectively (p < 0.001). CONCLUSION: The sensitivity of the membranous urethra decreases in patients after cystoprostatectomy and ileal bladder replacement. The sensitivity of the membranous urethra is better in continent patients than in incontinent patients and therefore appears to be an important factor for continence after ileal bladder replacement. PMID- 11400487 TI - [Results of an epidemiologic surgery carried out with men 50-80 years of age to study urinary disorders, quality of life and sexual function]. AB - OBJECTIVE: The prevalence of urinary symptoms, their impact on quality of life and sexuality and the man's attitude in relation to these problem were studied by a self-administered questionnaire (including I-PSS, 6 questions of DAN-PSS-1, BPHQL9 and IIEF). MATERIAL AND METHODS: This questionnaire was sent by mail to a national representative sample of 3,500 French men aged 50 to 80 years. Of the 2,372 men who returned an interpretable questionnaire, 21%, 33% and 42% belonged to the severe or moderate category for symptoms, tolerance of symptoms, and alteration of quality of life, respectively. RESULTS: 81% of men reported sexual activity during the last 4 weeks. 8.3% of men were treated with "antiprostate" medical treatments, and 8% had been operated (16% of them were treated medically after the operation). Only 29% and 17% of men had talked about their sexual and urinary problems, respectively. CONCLUSION: Alteration of functional and perceived sexuality was correlated with age and the severity of symptoms in non operated patients, but the poor correlations between scales evaluating sexuality and symptoms indicate that sexuality is a difficult aspect to investigate. PMID- 11400488 TI - [Multicenter study on dose escalation with conformal and conventional radiotherapy for the treatment of localized prostatic cancer. Preliminary results of tolerance and quality of life]. AB - OBJECTIVE: To study the safety of conformal radiotherapy dose escalation up to 80 Gy for curative treatment of prostate cancer. MATERIAL AND METHODS: A multicentre trial sponsored by the PHRC include 164 patients under the age of 75 years with stage T1b-T3 N0 M0 prostatic adenocarcinoma, between 1995 and 1998. The prostate was treated at 5 dose levels: 18 patients at 66 Gy, 28 at 70 Gy, 49 at 74 Gy, 48 at 78 Gy, 21 at 80 Gy. The acute and delayed toxicity was graded according to a multidisciplinary glossary. Quality of life was evaluated before and one year after treatment using the EORTC QLQ-C30 questionnaire and additional questions. RESULTS: 89% and 55% of mild or moderate gastrointestinal and urinary adverse effects were observed during treatment, respectively. At long-term follow-up, of the 162 evaluable patients, 12.3% presented grade 2 adverse effects and 2.5% presented grade 3 adverse effects (1 case of haematuria, 2 urethral strictures, 1 bladder neck lesion) with no significant difference between the various dose levels. Two successive quality of life questionnaires were available for 119 patients: tiredness, pain and dyspnoea increased in parallel, but non significantly in the groups receiving conventional doses or high doses. The proportion of patients reporting sexual activity without endocrine therapy decreased from 48% before treatment to 35% one year later. The proportion of patients with no clinical or laboratory signs of progression was higher in the high-dose group. CONCLUSION: No significant difference was observed between conventional dose levels and high dose levels in terms of toxicity or quality of life. As the adverse effects were acceptable, a randomized trial comparing 70 Gy and 80 Gy has been initiated. PMID- 11400489 TI - [Secondary biological recurrence after radical prostatectomy: multivariate analysis of prognostic clinical, biological, and histologic factors]. AB - OBJECTIVES: To define the clinical, laboratory and histological variables associated with secondary elevation, after an undetectable phase, of PSA after radical prostatectomy. MATERIALS AND METHODS: This was a prospective study of 83 consecutive patients undergoing retropubic radical prostatectomy with an undetectable postoperative PSA at 3 months. The predictive value of five preoperative criteria (age, total PSA, Gleason score on biopsies, positive apical biopsies, clinical stage) and four postoperative criteria (pathological stage, Gleason score on the operative specimen, positive margins, follow-up) for secondary laboratory progression was studied by univariate and multivariate analysis. RESULTS: With a median follow-up of 36 months, the laboratory recurrence rate was 19%. Laboratory recurrence was associated with a biopsy Gleason score greater than or equal to 7 (p = 0.04), a high pathological stage (p = 0.03), a high histological score (Gleason > or = 7) (p < 0.01) and positive margins (p = 0.04). Logistic regression identified a Gleason score on the operative specimen greater than or equal to 7 to be the only element predictive of secondary laboratory progression. CONCLUSION: The concept of positive margins or high pathological stage is insufficient to identify the risk of laboratory progression after radical prostatectomy. The Gleason score, which evaluates tumour aggressiveness, the risk of micrometastases or periprostatic extension, therefore appears to be more useful. PMID- 11400490 TI - [Periurethral collagen injections: results after 2 years in 25 patients with severe urinary incontinence]. AB - OBJECTIVES: To study the efficacy and safety of periurethral collagen injection in the treatment of severe urinary incontinence. PATIENTS AND METHODS: From 1994 to 1998, 25 periurethral collagen injections were performed in 25 women with a mean age of 65 years presenting with urinary incontinence classified as grade III in 17 cases and grade II in 9 cases, under local anaesthesia in 5 cases, spinal anaesthesia in 10 cases and general anaesthesia in 10 cases. Twenty patients were treated for recurrent incontinence. One injection was performed in 20 cases and two injections were performed in 5 cases. None of the patients presented disorders of pelvic muscles. The preoperative urodynamic assessment did not reveal any cases of detrusor instability, and showed a mean urethral closure pressure of 35 cm H2O. A skin test was performed in each case at least 30 days before the injection. RESULTS: The mean hospital stay was 2.75 days. The mean quantity of collagen injected was 5.7 ml. With a mean follow-up of 24 months, 33% of patients were continent, 39% were improved and 28% were considered to be failures. No complications were observed. CONCLUSION: Due to its safety, this method can be proposed in outpatients with good results, in patients of all ages, either as first-line treatment or for recurrent incontinence, as, in the case of failure, this technique does not comprise subsequent treatment by another operation. PMID- 11400491 TI - [Renal transitional carcinoma and caval tumor thrombus]. AB - Transitional cell carcinoma of the kidney with caval tumour thrombus is extremely rare. The authors describe the radiological signs suggesting the preoperative diagnosis and guiding the therapeutic approach. They present the case of a 51 year-old patient with excluded caliceal stones, and review 17 cases published over a period of 24 years during which considerable progress has been made in radiological investigations. Only CT appears to be able to indicate this aetiology and allows retrograde ureteropyelography looking for a tumour of the urinary tract. MRI provides the best assessment of tumour involvement of the inferior vena cava. Nephroureterectomy with excision of a perimeatal bladder cuff and cavotomy is the only oncological surgical procedure, despite the extremely poor prognosis. PMID- 11400492 TI - [Unusual form of adrenal pheochromocytoma: asymptomatic cystic mass]. AB - We report an adrenal cyst discovered in a 26-year-old woman with abdominal pain. Imaging (ultrasonography, computed tomography) demonstrated a 12 cm adrenal cystic mass suggestive of hydatic cyst of adrenal. She undergoes surgical exploration, when blood pressure increases. Pheochromocytoma diagnosis is reconsidered. Cystic pheochromocytoma is a rare tumour of the adrenal gland that can pose a diagnostic challenge. PMID- 11400493 TI - [Role of multiarray scanner in the choice of therapeutic strategy in kidney cancer. Report of a case of kidney tumor with double thrombus in inferior vena cava]. AB - Renal cancer with caval thrombus is a disease which requires precise preoperative assessment, in order to adapt the surgical technique to the stage and extent of the disease. Recent developments in CT imaging, spiral acquisition and, more recently, the multiarray technique, allow a high quality evaluation. In the light of a case of right renal tumour with bifocal caval extension, the authors discuss the valuable contribution of this imaging modality in the staging assessment and evaluation of arterial (single or multiple artery, anatomical situation in relation to the left renal vein, presence of early bifurcations, etc.), and venous structures (renal vein, genital vein, extension of the thrombus, lumbar vein and hepatic vein). Improvements in CT imaging constitute a major progress in the staging of renal cancers with caval thrombus and make a considerable contribution to the choice of treatment strategy. PMID- 11400494 TI - [Primary carcinoid tumor and horseshoe kidney: potential association]. AB - The authors present the ninth case of primary carcinoid tumour arising in a horseshoe kidney. The diagnosis and treatment were delayed due to the benign cystic appearance of the initial lesion. This exceptional association must be kept in mind, as horseshoe kidney is associated with an increased risk of malignant tumours, especially for carcinoid tumours. The minimally aggressive nature of these tumours generally allows limited surgical resection. PMID- 11400495 TI - [Transitional cell carcinoma on ureteral stump after nephrectomy for pyonephrosis]. AB - The authors report a case of transitional cell carcinoma of the left ureteral stump in a 66-year old man treated by nephrectomy for pyonephrosis 6 years previously and cystoprostatectomy for bladder tumour 13 years previously. In the light of this case and based on a review of the literature, they essentially discuss the diagnostic and aetiopathogenic problems raised by this disease. PMID- 11400496 TI - [Metastasis to trocar site after lymphadenectomy with retroperitoneal laparoscopy]. AB - The authors report a case of trocar site metastasis, one year after laparoscopic retroperitoneal lymphadenectomy. This is the seventh published case of trocar site metastasis in laparoscopic urological surgery. The lymph node resection procedure and the histological type of the tumour appear to be risk factors for the development of wall metastases. PMID- 11400497 TI - [Recurrent peri-stomal varix hemorrhage after diversion using the Bricker's technique]. AB - Peristomal varices can occur in patients with gastrointestinal or urinary diversions associated with portal hypertension. It is now no longer rare to propose radical surgery for invasive bladder tumours in patients also suffering from hepatic cirrhosis, responsible for specific subsequent complications. Less than ten cases of varicose haemorrhages have been described on ileal bladders. The authors report the case of a patient with cirrhosis (Child B) treated surgically (radical cystoprostatectomy and Bricker transileal cutaneous diversion) for invasive bladder tumour. Episodes of bleeding varices occurred two months after surgery. Repeated and abundant haemorrhage led to the placement of an intrahepatic shunt (TIPS) allowing reduction of the portal hypertension and the severity of the bleeding. When local control of the bleeding is no longer possible, reduction of the portosystemic pressure gradient is required. TIPS is an effective alternative to surgical shunts, responsible for high morbidity and mortality in these debilitated patients. PMID- 11400498 TI - [Urachal cyst. Report of 3 cases]. AB - The authors report three cases of cyst of the urachus, one of which was complicated by infection. This diagnosis is only rarely established clinically. The current performance of ultrasonography allows very precise analysis of the anterosuperior surface of the bladder, which may reveal the diagnosis. The authors discuss the problems raised by cysts of the urachus and emphasize the importance of clinical history, physical examination and complementary investigations, particularly CT which should allow the diagnosis of this disease, which needs to be treated surgically. PMID- 11400499 TI - [Post-traumatic complication of trans-appendiceal cystostomy: urinary peritonitis]. AB - Two children who had undergone a transappendicular urinary diversion (type Mitrofanoff) developed bladder rupture, one following abdominal trauma, 4 months after the operation and the other following traumatic self-catheterization at 4 years. The clinical history and standard radiological examinations (ultrasonography, cystography) confirmed the diagnosis of urinary peritonitis. Emergency surgical repair was possible in both cases with an uneventful postoperative course. This serious and rare complication requires emergency surgery and justifies rigorous selection of children suitable for this type of diversion giving preference to increased bladder neck resistance over bladder neck closure. PMID- 11400500 TI - [Pulmonary thromboembolism after chronic bladder distention]. AB - OBJECTIVE: To report a case of pulmonary embolism secondary to chronic over distended bladder, responsible for iliac vein obstruction. CLINICAL CASE: A 73 year-old patient presented with sudden onset of pulmonary embolism, confirmed by isotope lung scan, after drainage of 2,300 cc of urine from an over-distended bladder, responsible for bilateral iliac vein obstruction and a bilateral obstructive uropathy. The patient had a favourable course after anticoagulation with heparin and temporary bladder catheterization, leading to resolution of the peripheral oedema and obstructive uropathy. CONCLUSION: About thirty cases of obstruction of large pelvic vessels due to over-distended bladder have been reported to date. However, and after an extensive review of the literature, this case appears to be the first report of pulmonary embolism secondary to chronic over-distended bladder. PMID- 11400501 TI - [Sarcomatoid carcinoma of the prostate]. AB - The authors report a case of sarcomatoid carcinoma of the prostate in a 49-year old patient. The clinical symptoms were dominated by dysuria and frequency. Digital rectal examination showed a very large, hard and irregular prostate. Serum PSA was normal. Transrectal ultrasonography and abdominopelvic computed tomography showed a heterogeneous mass occupying the vesico-prostatic junction. The patient underwent cystoprostatectomy. Histological examination of the cystoprostatectomy specimen revealed a tumour composed of spindle cells expressing epithelial markers on immunohistochemical analysis and concluded on a diagnosis of sarcomatoid carcinoma of the prostate. The patient died seven months after cystoprostatectomy. PMID- 11400502 TI - [A case of recto-urethral fistula in Crohn's disease]. AB - The authors report the fourteenth case of rectourethral fistula associated with Crohn's disease, in a 53-year-old man with a 20-year history of ileocolonic Crohn's disease. The patient developed a rectourethral fistula following an episode of complex anoperineal suppuration surgically exposed on several occasions. Repeated surgical exposure and urine drainage were not sufficient to close the fistula and cure the perineal suppuration. After 10 months, the patient was treated by the rectal mucosal transposition flap technique to close the rectal aspect of the fistula, which ensured cure of the patient's fistula and anoperineal lesions. This technique has been used successfully on 4 occasions and appears to be an effective treatment for rectourethral fistula in Crohn's disease with healthy rectal mucosa. PMID- 11400503 TI - [Harpoon scrotal injury: unusual accident in submarine fishing]. AB - The authors report a case of transfixing injury of the scrotum while diving. The patient was seen one hour after the accident. The harpoon traversed the right side of the scrotum from below upwards. Emergency surgical exploration only revealed effraction of the sinus of the epididymis and the abdominal muscles. The overall risk for fertility was suggested in view of a possible lesion of the epididymal canal. PMID- 11400504 TI - [Treatment of stress urinary incontinence with percutaneous colposuspension: unsatisfactory technique]. AB - OBJECTIVE: The objective of this study was to evaluate the results of a percutaneous needle colposuspension technique with bone fixation (Vesica system) in the treatment of female urinary stress incontinence (USI) of the woman, with a minimum follow-up of one-year. MATERIAL AND METHODS: 34 women aged 35 to 86 years (mean: 62 years) were treated for USI due to bladder neck hypermobility according to the Vesica technique. All patients had a positive preoperative Bonney manoeuvre without sphincter incompetence on urethral pressure profile. RESULTS: Overall, 1 year postoperatively, 9 patients (26%) were completely dry and did not wear any protection, 19 (56%) were improved, but still presented occasional leaks and 6 (17%) were considered to be failures. Physical examination revealed postoperative leaks in 24 patients with recurrence of hypermobility and positive Bonney manoeuvre in every case. There was one subcutaneous abscess and 2 cases of persistent pain at the bone implant site. No cases of bone infection or screw explantation were observed. No cases of retention beyond the 3rd postoperative day were observed. CONCLUSION: The percutaneous colposuspension technique gives disappointing results at one year due to recurrence of urethral hypermobility in every case. PMID- 11400505 TI - [Cure of cystocele with vaginal patch]. AB - OBJECTIVE: To describe an original technique for transvaginal cystocele repair and to report the anatomical and functional results of the first 47 patients operated by this technique. The operation combines suspension, by six sutures to the tendinous arches of the pelvic fascia, of a vaginal patch measuring 6 to 8 cm long by about 4 cm wide left in contact with the cystocele. The vaginal patch is then buried under the anterior colporraphy suture. Spinal fixation, hysterectomy, levator myorrhaphy or urinary incontinence repair are also performed as necessary. METHODS: Descriptive retrospective study of 47 patients undergoing transvaginal repair of prolapse between October 1997 and June 1998. All patients presented external prolapse with grade III cystocele, associated with urinary incontinence in 38.3% of cases and hysterocele or vaginal prolapse in 87% of cases. The mean age of the patients was 69 years. The uterus was preserved in two cases, but hysterectomy was performed in the other 45 patients, together with Richter spinal fixation in 44 cases An associated levator myorrhaphy was performed in every case. RESULTS: The mean follow-up was 16.4 months (range: 6 to 26 months) in 46 patients. The cystocele was considered to be cured in 93% of patients, one patient had an asymptomatic grade I cystocele and surgery was unsuccessful in two patients who developed recurrent grade II cystocele. CONCLUSION: The technique presented here is a curative treatment for grade III cystocele in menopaused women associated with Richter spinal fixation, and prevents the risk of cystocele described after transvaginal treatment of prolapse by spinal fixation alone (10 to 20%). The short-term results are encouraging, but they need to be confirmed by a follow-up of at least 5 years. PMID- 11400506 TI - [Economic comparison of 2 surgical techniques fr the treatment of stress urinary incontinence in women: Burch's technique versus the TVT technique]. AB - The TVT device (Tension-free Vaginal Tape) is used in our Hospital to treat stress urinary incontinence, resulting in an excess cost for the Pharmacy. The Burch technique, used previously, does not require any specific medical device, but is invasive and requires a longer hospital stay. The objective of this study was to compare the financial impact of these two techniques, by defining the discriminant costs. Seventeen isolated Burch procedures and twenty one TVT procedures were included. The costs analysed concerned medical devices, medicinal products, laboratory procedures, operating time, hospital stay and duration of postoperative follow-up. The Burch procedure cost FFR 26,322 and the TVT procedure cost FFR 10,958. The TVT technique reduces the cost of hospitalisation and represents an economy of operative equipment and nursing workload (reduction of operating time and postoperative stay). PMID- 11400507 TI - [Syringocele: report of 2 cases]. AB - Cowper's syringocele, dilatation of bulbo-urethral glands, is a rare disease, usually congenital. The authors report two cases of perforated syringocele and analyse the clinical, radiological and therapeutic aspects of this disease. PMID- 11400508 TI - [Testicular microlithiasis and cryptorchism: ultrasound analysis after orchidopexy]. AB - OBJECTIVE: To evaluate the testicular ultrasound features and frequency of testicular microlithiasis in a population of patients operated several years before for cryptorchidism, in order to define a group at possible increased risk of testicular cancer. MATERIAL AND METHODS: This was a retrospective study of 202 patients, 63 of whom were reviewed by ultrasound with a mean follow-up of 9 years 3 months (7 years 5 months to 11 years 7 months). RESULTS: 32% of operated testes were hypotrophic compared to the correctly descended side; their mean volume was 9.42 ml versus 11 ml for the nonoperated side. 14.3% of surgically descended testes presented an ultrasonographically heterogeneous parenchyma and microlithiasis were present in 9.52% of cases. CONCLUSION: The association between microlithiasis and cryptorchidism is not an incidental finding. Apart from the relative hypotrophy of the testis, ultrasound also revealed, in our study, that the presence of microlithiasis is not an extremely rare finding. Annual ultrasound surveillance is therefore required due to the risk of malignant transformation of these microlithiases. PMID- 11400509 TI - [Diagnostic and therapeutic standards in adrenal tumors]. PMID- 11400510 TI - [A necessary number or subjects... to conclude]. AB - The fact that a difference is not statistically significant does not mean that it does not exist. Some differences are not demonstrated by studies conducted on small sample sizes due to random sampling fluctuations. To demonstrate a difference which really exists or to conclude on the absence of a difference, the study must have a sufficient power, essentially determined by the number of subjects included. In the case of comparisons of survival curves, the number of events or total deaths at the time of analysis determines the power of the study. Examples derived from the urological or theoretical literature will illustrate the value of calculating the number of subjects required before starting the study and the value of retrospectively evaluating the power of a study in order to interpret its results. PMID- 11400511 TI - [Interleukin-6 and bone metastasis of renal cancer: molecular bases and therapeutic implications]. AB - Interleukin-6 (IL-6) is a multifunctional cytokine which provides multiple signals on various tissues and cells. In addition, IL-6 is produced by some human renal carcinoma cell lines in vitro and is expressed in a majority of primary renal cell carcinoma (RCC). Serum IL-6 influence the response to immunotherapy. IL-6 appears as a target for rational drug design highly promising for development of new cancer therapies. IL-6 effects are mediated by Stat. Stat (Signal transducers and activators of transcription) signaling pathways represent novel molecular targets for therapeutic implications in metastatic renal cell carcinoma. Inhibitors of Stat signaling pathway will not only block tumor growth by inducing apoptosis, but may also increase the sensitivity of tumors to conventional treatment (immunotherapy). The processes involved in tumor associated angiogenesis should lead to compounds able to interfere with angiogenesis. Il-6 has been implicated in the osteoclastic bone resorption and hypercalcemia associated with metastatic RCC. Different agents were shown to be effective in treating lytic bone disease mediated by osteoclast activation: bisphosphonates and osteoprotegerin. PMID- 11400512 TI - [Giving a future to clinical (urologic) research in France. Point of view of the medical director in the pharmaceutical industry]. AB - The regulation of clinical research in France is aimed essentially at the protection of individuals and the maintenance of good clinical practice. However, in an international context, certain obstacles linked to this regulation, as well as other obstacles of a financial and organisational nature, limit the participation of the French investigational centers, particularly in private clinical research. Legal texts and practical concerns are considered in this light; solutions are outlined. PMID- 11400513 TI - [Transverse tubulization of intestinal segments: a catheterizable conduit as an alternative to the Mitrofanoff procedure]. AB - This article describes a procedure for creation of an intestinal conduit (that remains available for a continent diversion) by transverse tubulisation of intestinal segments. This simple, effective technique represents an alternative to Mitrofanoff's procedure in patients with a non-viable appendix. PMID- 11400514 TI - [A new procedure based on de-epithelialization in the treatment of scrotal hypospadias]. AB - OBJECTIVES: To prevent the development of postoperative urethral fistula and to correct bifid scrotum. MATERIALS AND METHODS: Eleven boys with scrotal hypospadias were treated by this technique, which consists of raising a vascularized skin flap from the internal surface of each hemiscrotum, which is then de-epithelialized and closed over the urethroplasty. RESULTS: Two of the 11 patients developed fistulas that resolved spontaneously with a better cosmetic appearance of the scrotum. CONCLUSION: This technique appears to be very effective in the prevention of postoperative urethral fistula, and also allows correction of bifid scrotum. PMID- 11400515 TI - [Integration activities of a central municipal hospital]. AB - Scientifically-based integration of therapeutic and prophylactic institutions is needed for better meeting the population requirement of medical care, and it is desirable to unite the financial resources and personnel; such is the objective reality for municipal therapeutic and prophylactic institutions of local public health systems. In order to make medical care available for the population and to ensure its high quality, structural changes in the public health of a region are needed. These changes include integration of medical services of these territories; creation of a network of therapeutic and prophylactic institutions corresponding to medical demographic structure of population, so that the scope of medical care be increased and specialized care made easier available for the population; and creation of the optimal managing system. Comparative studies of population health and its time course, public health organization at neighboring territories, where therapeutic and prophylactic institutions, such as Central Municipal Hospital and Central Regional Hospital have autonomous managing and financing, confirmed the need in integration of public health units functioning at a certain territory into a universal system, and in development of approaches to overcome the present-day miscellaneous network by integrating the activities of treatment-and-prophylaxis institutions. Identical economic, geographic, and macroeconomic living conditions of the population, similarity of medical demographic structure and similar changes in it, as well as similar morbidity structure, are sufficient grounds for integration of public health services. PMID- 11400516 TI - [Development of monitoring of population health, environment and public health at a regional level]. AB - A structural model of monitoring, intending the creation of a multifunctional multi-level information system, is suggested for creation of a universal information analytical data bank on population health, human environment, and public health activities at a regional level. PMID- 11400517 TI - [Structure of medical work and its evaluation: methodological aspects]. AB - Methodological approaches to studies and description of the structure of medical work are presented with the aim of evaluating it. Studies of labor structure should result in the formation of a complex of its parameters (properties) to be taken account of, which is the priority task in developing a system of evaluations. Medical labor is regarded as a complex integral system requiring a systemic approach. The methodology is based on qualimetric principles and methods. The fundamental principles are necessity and sufficiency. The method for evaluation of medical work is to correspond to the actual potentials and routine of medical institutions. PMID- 11400518 TI - [Evaluation of the efficiency of the activities of therapeutic and preventive services]. AB - Two methodological approaches to evaluation of the efficiency of medical activities, based on estimation of an integral index and expert evaluations, were investigated in 14 hospitals of Chelyabinsk. Both approaches are based on 12 parameters characterizing the work of outpatient clinics and hospitals and are routinely used in medical statistics. Comparison of classifications of treatment and-prophylaxis institutions of Chelyabinsk in 1994-1998 obtained with the use of both methods showed that the method based on estimation of the integral parameter is more objective. Its another advantage is the possibility to modify the number of signs used and thus to evaluate the efficiency of the entire medical institution and its structural units (outpatient center, hospital). The efficiency rating can serve as the basis for adopting appropriate managing decisions. PMID- 11400519 TI - [Features of medical legislation]. AB - Presents the major characteristics of medical legislation as a legal unit. Special attention is paid to manifestations of common principles of systemic legislation and signs characteristic of specialized laws. PMID- 11400520 TI - [Relations between the problem-oriented education of patients and quality of treatment and efficiency of rehabilitation of children with chronic diseases]. AB - Research aimed at development and introduction of effective forms of organization and methods of health improvement, compensation, and rehabilitation of patients with chronic diseases acquire special significance today. However, the actual economic and social conditions in the Russian Federation dictate the development and introduction of the least expensive and at the same time sufficiently effective methods for treatment and rehabilitation. Topical training of chronic patients is such a method. One more aspect necessitating such research is essential increase in the prevalence of chronic diseases and disability among children. Special attention is paid to insulin-dependent diabetes mellitus, asthma, and chronic gastrointestinal diseases, often leading to stable (sometimes for life) disability; treatment, rehabilitation, and follow-up of such patients are very expensive, which makes the problem a pressing one. PMID- 11400521 TI - [Biomedical and social-hygienic risk factors of disability in rural children]. AB - The importance of the problem of disability of children causes no doubts. The author defines the term "inability" which is recommended for wide use. The data of comprehensive studies of the life style and life conditions of the rural families with disabled children are summed up. The totality of factors affecting a child's health is analyzed. Subjective requirements of children with limited potentialities in medical care are investigated. Probable approaches to decrease of children's disability are discussed. PMID- 11400522 TI - [Current problems of delivering medical care on ships]. AB - Medical personnel of ships has been liquidated during the period of public health reforms, and in accordance with International Regulations of Floating Safety, sailors now have to be trained to render first medical care and care of victims on board the ship. This paper discusses the problems essential for such training of sailors and presents the experience gained by the first worldwide department of marine medical training of sailors at S. O. Makarov State Navy Academy. Basic characteristics of navy legislation are presented. Special attention is paid to general laws and laws essential for this sphere. PMID- 11400523 TI - [Problems in teaching population health and public health]. PMID- 11400524 TI - [Legal support of public health development in the Murmansk region]. PMID- 11400525 TI - [Contracts and mutual accounts in therapeutic and preventive institutions]. PMID- 11400526 TI - [Public health reforms in Ukraine: legal bases]. AB - The authors demonstrate the need in a law of the Ukraine on organization of population health protection by using an original methodological approach: they separate organization of medical activity and primary health care. The state is expected to play the leading role in public health as a regulator, but not manager. Philosophy of financial/economic and organization/financial basis of medical activities is described in the context of modern legislation. PMID- 11400527 TI - [Changes in the organization and content of training in connection with the "Additional Resolution on the Medical Department of the Imperial Moscow University" (1845)]. PMID- 11400528 TI - [Military hospitals in Russia at the end of the 19th and beginning of the 20th centuries]. PMID- 11400529 TI - [Legislative acts as a source of research in history of medicine]. PMID- 11400530 TI - [The Botkin Commission (1886-1889)]. PMID- 11400531 TI - [Main trends in the activities of the Medical Department of Ministry of Internal Affairs in the second half of the 19th century]. PMID- 11400532 TI - [History of scientific medical societies in Saratov]. PMID- 11400533 TI - [Therapeutics in the second half of the 18th century--beginning of the 19th century]. PMID- 11400534 TI - [Folk medicine in the Vyatka province in the 18th century--beginning of the 20th century]. PMID- 11400535 TI - [Public health in Belarus in the 1930-ies]. PMID- 11400536 TI - [Specific features of the incidence of ischemic heart disease in Dagestan]. AB - The authors analyze the culture, socio-economic and communal characteristics of life style in Daghestan, determining the population health in this republic. Recommendations on developing the strategies for prevention of coronary disease in the Republic of Daghestan are offered. PMID- 11400537 TI - [Marriage in old age groups (medico-social aspects)]. AB - Recently married elderly people change attitude to hospital treatment and living in asylums; they prefer care rendered at home and hospital-substituting forms of care; hospitalization and ambulance care are less needed and permanent social workers are required less. All this promotes rational utilization of the potentialities of therapeutic institutions and saving of public health and social protection resources. PMID- 11400538 TI - Adult patients with treated complete cleft lip and palate. Methodological and clinical studies. AB - The purpose of the present thesis was to investigate the quality of life, satisfaction with treatment, prevalence of temporomandibular disorders, psychosocial distress, and occlusal stability in a treated group of adults with complete cleft lip and palate (CLP). Sixty-eight adults (44 men and 24 women) with a mean age of 24.2 years (range 19.5-29.2) with treated CLP were compared with a gender- and age-matched group with no clefts. The CLP subjects were born between 1968 and 1977 and had undergone standardised plastic surgery at the Department of Plastic Surgery, University Hospital, Linkoping, Sweden. Logopaedic, phoniatric, otological, and orthodontic examinations and treatment had been provided locally, supervised by the Cleft Plate Team. The subjects answered a multidimensional, self-report, standardised questionnaire regarding psychological and somatic conditions. The subjects underwent a clinical TMD examination and an evaluation of the occlusion. The reliability of the multidimensional questionnaire was analysed for the CLP group by a test-retest study within a 2-3 week interval and most questions showed an overall good reliability. A panel of professionals judged the outcome of the surgical treatment on colour slides of the CLP subjects. The dental plaster casts of 39 subjects born with complete unilateral cleft lip and palate (UCLP) were analysed (mean age 24.7 years, range 20.2-29.3) and compared with the dental plaster casts taken at mean age of 19.1 years (range 16.0-20.6). The overall level of quality of life was rather high in both groups. The CLP group rated some detached aspects, such as life meaning, family life, and private economy, significantly lower than did the group without clefts. Overall aspects such as well-being and social life were affected by having a treated cleft but not the more practical and tangible aspects of their daily living. There was an overall high level of satisfaction with all the different part of the body in both groups, but the CLP group reported significantly more dissatisfaction with their nose, lips, mouth, profile, and overall facial appearance than the group without clefts. The professionals and the subjects with CLP were generally not very satisfied with the results of surgical treatment. Thirty of the subjects with CLP (47%) wished to have more operations. The professional group recommended further operations in 38 of the subjects (59%) in particular, rhinoplasties. The CLP group had significantly higher frequencies of cross-bite than the group without clefts, but no differences regarding TMD pain were found between the two groups. In the subjects with treated UCLP, there was a significant deterioration in the occlusal score and the maxillary arch dimensions between 19 and 25 years. This was irrespective of the type of retention. The persisting morphological malocclusion with a low frequency of interferences has had no influence on TMD symptoms in the group of CLP patients studied. The conclusion is that the CLP subjects in the present study seemed to be psychosocially well adjusted to their disability. However, 47 per cent wished to have further surgical treatment. The persisting malocclusions did not provoke TMD symptoms. PMID- 11400539 TI - Pop culture: dentists decry soda in schools. PMID- 11400540 TI - Biofilms. An introduction. PMID- 11400541 TI - Bacterial biofilm and dentistry. AB - Bacterial biofilms are ubiquitous in nature. Recent studies have demonstrated many unique qualities previously unknown to bacteria and have yielded new insights into relevant dental issues. PMID- 11400542 TI - Periodontitis as a biofilm infection. AB - Microbial biofilms are a major concern of infections associated with implantable medical devices as well as with many non-implant related chronic infectious diseases in humans. Dental plaque is also a biofilm. Dental plaque is probably one of the best characterized biofilms occurring on the surface of human tissues. This article will examine the impact of biofilm research on concepts of microbial etiology and the treatment implications for periodontitis. PMID- 11400543 TI - Modern anti-HIV therapy. AB - Highly active antiretroviral therapies (HAART), usually consisting of two nucleoside reverse transcriptase inhibitors (NRTI) plus an HIV protease inhibitor (PI), have been widely used since 1996. They produce durable suppression of viral replication with undetectable plasma levels of HIV-RNA in more than half of patients. Immunity recovers, and morbidity and mortality fall by more than 80% [1, 2]. Treatment was thought to be particularly effective when started early; therefore, HAART was recommended for essentially all HIV-infected persons willing to commit themselves to lifelong therapy [3, 4]. Besides these successes, however, HAART also produces problems. HIV is not eradicated by present-day drugs, and patients often cannot comply with long-term combination treatment [5, 6]. Moreover, HAART causes unexpected and ill-understood side effects [7]. The dogma of earliest possible treatment has therefore come under attack. Ten principles governing anti-retroviral treatment are summarised in Table 1. Starting and maintaining HAART is complex. Within the last few years, the numbers of antiretrovirals, their known and potential interactions with each other and with non-HIV drugs, and the list of their side effects have all increased exponentially. As a rule a physician specialising in HIV care should be consulted whenever HAART is started or changed. It is his task to ensure that the treatment chosen is optimal for the particular patient. PMID- 11400544 TI - Post-prandial lipaemia and endothelial function among healthy men. AB - BACKGROUND: There is evidence that elevated post-prandial lipoproteins adversely affect progression and outcome of cardiovascular disease. Traditional risk factors are associated with impaired endothelium-mediated vasodilatation. However, studies regarding the relationship between post-prandial lipaemia and endothelial function are divergent. METHODS: Twelve healthy non-smokers were included in this study. Before and after intake of a lipid cocktail rich in dairy fat, we tested endothelial-dependent (acetylcholine 0.8-160 mg/min per 100 ml forearm tissue) and -independent (sodium nitroprussid 0.6 microgram/min) vascular function in the forearm vascular bed with plethysmography. Moreover, we tested the effect of 1-NMMA, a competitive inhibitor of the NO synthetase, on base-line flow. Extent of post-prandial lipaemia was assessed with the increases in triglycerides and retinyl-palmitate, a marker for intestinally derived lipoproteins. RESULTS: Baseline flow was higher after the test meal than during fasting (preprandial 6.5 +/- 0.5 ml/min* 100 ml tissue, post-prandial 8.0 +/- 0.5, p = 0.03), but similar after 1-NMMA (p = 0.85). Before and after intake of the test meal, there was no significant difference in acetylcholine-induced endothelium-dependent vasodilatation (repeated measurement ANOVA, p = 0.22). At the highest acetylcholine dose, forearm flow was very similar (fasting 18.4 +/- 1.9, post-prandial 17.9 +/- 1.9, p = 0.75). At maximum acetylcholine dose, there was a weak inverse but non-significant correlation between forearm flow and post prandial triglyceridaemia (r = -0.38, p = 0.23) and intestinally derived lipoproteins (chylomicrons r = -0.29, p = 0.35, chylomicron remnants r = -0.15, p = 0.63). However, at the lowest acetylcholine dose there was a suggestion for a positive correlation between change in flow and post-prandial lipaemia (triglyceridaemia, r = 0.53, p = 0.07; chylomicrons, r = 0.41, p = 0.18 and remnants, r = 0.51, p = 0.09). Endothelium-independent vasodilatation in response to sodium nitroprusside did not significantly change (p = 0.23). CONCLUSION: Our results suggest that among healthy men post-prandial lipaemia is not associated with a notable impairment of endothelium-mediated vascular function in forearm resistance vessels. PMID- 11400545 TI - Evaluation of the FASTSTART mode for reducing start-up delay in syringe pump infusion systems. AB - OBJECTIVE: The aim of the study was to evaluate the IVAC P7000 FASTSTART mode with regard to start-up performance in a 50-ml infusion syringe at a flow rate of 1 ml.h-1. METHODS: The time from depression of the start button to first fluid flow (T1) and to establishment of a pre-set flow rate (T2) were gravimetrically recorded with and without FASTSTART and with and without priming of the infusion system with a 1-ml fluid bolus prior to connection of the infusion line to the patient. RESULTS: FASTSTART significantly reduced start-up times in the unprimed syringe pump infusion system from (mean [SD]) 9.4 (6.0) to 2.5 (3.5) min for T1 and from 21.8 (9.8) to 9.4 (6.2) min for T2 (all p < 0.001). The greatest improvement in shortening of T1 and T2 was obtained when the system was primed prior to starting (p < 0.0001). After priming the infusion system, FASTSTART shortened T2 by some 50% from 1.4 (1.4) to 0.7 (0.6) min. CONCLUSION: Our data indicate that the FASTSTART procedure is effective and that substantial improvements can be obtained by priming the system prior to starting. PMID- 11400546 TI - BRCA1/2 mutations in Swiss patients with familial or early-onset breast and ovarian cancer. AB - QUESTIONS UNDER STUDY: Germ-line alterations in BRCA1 and BRCA2 genes account for 30-50% of all forms of familial breast and ovarian cancer syndromes. Specific mutations in specific populations and ethnic groups have been identified in BRCA1 and BRCA2. However, it is not known whether such specific mutations prevail in the Swiss population. METHODS: We started to screen patients with primary breast and ovarian cancer and a strong family history of both cancers by sequencing the full-length coding regions of BRCA1 and BRCA2. RESULTS: With the selection criteria used in this study we identified 19 mutations in the first 38 patients screened (50%). These mutations were either defined as deleterious and resulted in a protein truncation (n = 10) or were defined as unclassified variants (n = 9). One novel truncating mutation was found in BRCA2 and two novel unclassified variants were detected in BRCA1. These three mutations are not described in the BIC and HGMD databanks. CONCLUSIONS: We detected three unknown mutations among 38 patients in a Swiss study of BRCA1/2 mutation patterns. One of these novel mutations is clearly deleterious as it leads to protein truncation at nucleotide 133 of BRCA2. PMID- 11400547 TI - Three cases of severe neurotoxicity after cobra bite (Naja kaouthia). PMID- 11400548 TI - Progress in the separation of enantiomers of chiral drugs by TLC without their prior derivatization. PMID- 11400549 TI - Bis-naphthylureas and related compounds: synthesis, chemical properties, DNA affinity and antineoplastic activity. AB - A series of bis-naphthalene derivatives in which these moieties are linked by a symmetric bis-urea functionalized chain or by an asymmetric amide and urea or amino and urea functionalized chain, were synthesized. The tentative synthesis of other types of related compounds did not give the products expected. The compounds were assayed as antineoplastics on human tumor cell lines at the National Cancer Institute (USA). Bis-urea derivatives were active on lung, colon, renal and CNS tumor cell lines. The degree of affinity of these compounds to DNA was also studied, showing low affinity. PMID- 11400550 TI - Possible antineoplastic agents, part 15: synthesis, biological activity and quantitative structure activity relationship of substituted-2-(4' methoxybenzenesulphonamido) glutaric acid analogs against Ehrlich ascites carcinoma. AB - Some substituted-2-(4'-methoxybenzenesulphonamido) glutaric acid analogs (5a-m, 7a-d) have been synthesized and tested for their possible antineoplastic activity against Ehrlich ascites carcinoma (EAC) in Swiss albino mice using tumour (ascitic fluid) weight as activity parameter. Some of these compounds possess encouraging antitumour activity. A QSAR study, performed by the classical Hansch method, explains the significance of hydrophobic binding and electronic influence in the mechanism of antineoplastic action in this group of compounds. PMID- 11400551 TI - Synthesis and antitumor activity of some new phthalimide analogues. AB - The reactive intermediate 1-(chloroalkyl)-1-aza-2-azoniaallene salts 3, which were prepared by treatment of the alpha,alpha'-dichloroazo derivatives 2 with SbCl5, underwent cycloaddition with the N-cyanoalkyl phthalimide compounds 4 and afforded the 1,2,4-triazolium salts 5. These salts rearranged spontaneously to the protonated 1,2,4-triazoles 6, followed by hydrolysis in situ to the 1,2,4 triazolo-alkyl-phthalimide compounds 7. The newly synthesized compounds were then evaluated for their antitumor activity in three cell lines. PMID- 11400552 TI - Synthesis of heterobicyclic nitrogen systems bearing the 1,2,4-triazine moiety as anti-HIV and anticancer drugs, part III. AB - Some new fused heterobicyclic nitrogen systems such as 1,2,4-triazino[3,4-b] [1,3,4] thiadiazolones/thiadiazinones 4-15 and the related compounds 16-21 have been synthesized from treatment of 4-amino-3-mercapto-6-substituted-1,2,4-triazin 5-ones 1 with bifunctional oxygen and halogen compounds via heterocyclization reactions. Structures of the products have been deduced from their elemental analysis and spectral data. Significant anti-HIV and anticancer activities were observed in vitro for some members of the series, compounds 1e, 4e, 4f, 5, 6 and 16 showing a significant activity in Leukemia, Lung, Breast and CNS anticancer evaluation. PMID- 11400553 TI - Synthesis of some pyrrolo[3,4-d]pyridazinones and their preliminary anticancer, antimycobacterial and CNS screening. AB - Starting from 1-substituted-2,5-dimethyl-3,4-pyrroledicarboxylic acid anhydrides 1 and N-methylhydrazine, 1,2,3,4-tetrahydro-6-substituted-2,5,7-trimethyl-6H pyrrolo[3,4-d]pyridazin- 1,4-ones 2 were prepared. Reaction of compounds 2 with alkylating agents give 3-N- or 4-O-alkylated products 3, 4 or mixtures of these isomers in ratios depending on the alkylating agents. Under preliminary pharmacological screening two of four new pyrrolo[3,4-d]pyridazinones were active as cystostatic agents, all the eight compounds displayed moderate activity against Mycobacterium tuberculosis and two compounds were active as CNS depressive agents. PMID- 11400554 TI - Cytotoxic and anticancer properties of some 4-aryl-3-arylcarbonyl-1-ethyl-4 piperidinols and related compounds. AB - A previous investigation revealed that various 4-aryl-3-arylcarbonyl-1-ethyl-4 piperidinols and related vinylogs were cytotoxic to both murine and human tumour cell lines. In particular, 1a and 2a were identified as useful prototypic molecules. Structural modifications of 1a and 2a were accomplished leading to 1b e and 2b-d which displayed cytotoxicity towards murine P388 and L1210 leukemic cells as well as human Molt 4/C8 and CEM T-lymphocytes. Among the new compounds, the greatest average potencies against these four cell lines were displayed by 1b and 2b, having approximately one quarter and one half of the potency of the reference drug melphalan, respectively. The synthesis and bioevaluation of three open chain analogues of 1b-d, namely 3a-c, did not reveal unequivocally whether this molecular modification led to increases in cytotoxicity or not. Compounds 2a d were substantially more active than melphalan using a panel of human tumour cell lines. In addition, several compounds displayed selective toxicity to both colon and leukemic cancer cells. The 4-piperidinol 2d was active in the in vivo hollow fibre assay. This study revealed compounds with greater potency than 1a and 2a and it has confirmed that 1,3,4-trisubstituted-4-piperidinols and related compounds are novel groups of candidate antineoplastic and anticancer agents. PMID- 11400555 TI - Analysis of ubidecarenone (CoQ10) aqueous samples using reversed phase liquid chromatography. AB - A novel method was developed for the quantitative determination of ubidecarenone (CoQ10) in aqueous media using non aqueous reversed phase liquid chromatography. Standards were prepared by melting the compound in Cremophor EL followed by dilution with distilled water. Samples were then analyzed by a Reverse Phase HPLC method using a Waters Novapak C18, 3.9 x 150 mm column. The mobile phase used was methanol: n-hexane (9:1) at a flow rate of 1.5 ml/min. Response of the detector to the analyte was linear (r: 0.999) over the range of 2.5-100 micrograms ml-1, with a limit of detection of 0.17 microgram ml-1. Acetone used to solubilize CoQ10 in the surfactant and Cremophor EL did not interfere with sample analysis. This method provided a convenient and alternative approach to the existing methods that require organic solvent extractions prior to analysis. Besides, this method enabled the separation of major photolytic decomposition products of CoQ10. PMID- 11400556 TI - Column-switching chromatographic determination of itraconazole and its metabolite hydroxy-itraconazole in human serum. AB - A column-switching HPLC assay is described for the determination of the antifungal agent itraconazole and its main metabolite hydroxy-itraconazole in serum samples. Three precolumns packed with alkyl-diol silica sorbents differing in alkyl chain length are compared in the sample clean-up step. Chromatographic separation is achieved with a Symmetry C8 column. The assay presented shows a robust and selective analytical procedure with low requirements for sample quantity, no manual sample treatment and high sample throughput. PMID- 11400557 TI - [Color reactions for identification of sodium cromoglycate]. AB - Chromone-2-carboxylic acid (3) reacts with aminopyrazolone (2) in methanolic hydrochloric acid to yield the orange-red polymethine dye 4. Treating dimethyl cromoglicate (6) with compound 2 and perchloric acid leads to the tetraperchlorate 7 of the red azamerocyanine 8. The phenol 9, obtained from alkaline hydrolysis of sodium cromoglicate (1a), couples with diazotized sulfanilic acid to form the red azo dye 10. The chromone 6 condenses with 1,3 dimethylbarbituric acid (DMBA) in acetic anhydride/acetic acid to the red oxonole 12. Cromoglicinic acid (1b) reacts under these conditions to yield the yellow polymethine dye 14, whose structure is elucidated by X-ray analysis. PMID- 11400558 TI - Prediction of Caco-2 cell permeability using partial least squares multivariate analysis. AB - The permeability across Caco-2 cell monolayers of structurally diverse compounds were predicted using computed molecular descriptors and multivariate. Partial Least Squares (PLS). The molecular descriptors including log polarization, log solvent accessible surface area, hydration energy, heat of formation, and dipole moment were calculated with Hyperchem and ChemPlus QSAR programs for Windows. Other physiochemical properties such as hydrogen acceptor for oxygen atoms, hydrogen acceptor for nitrogen atoms, hydrogen bond donors, hydrogen bond forming ability, molecular weight, and log distribution coefficient were also used as descriptors. Cross validation with internal test set and prediction with external test set indicated the usefulness of the derived model for Caco-2 cell permeability. Hydrogen bonding is one of the important factors associated with permeability. While increased logD and hydration energy facilitate permeability, an increased dipole moment of molecules has a negative effect on permeability. PMID- 11400559 TI - Evaluation of mutagenic activity of several antimalarial extracts from Eupatorium inulaefolium. AB - Eupatorium inulaefolium is used as an antimalarial agent by traditional healers of the Tumaco region (Narino-Colombia). Several extracts of this plant have been tested by our laboratory and in vitro antimalarial activity against the FCB-2 strain of Plasmodium falciparum has been confirmed. For this reason, the mutagenic effect of the methanol, dichloromethane, and hexane extracts of Eupatorium inulaefolium (number 83377 university of Antioquia herbarium) were evaluated using the Ames test. None of the extracts evaluated had mutagenic effects on TA-98 or TA-100 strains of Salmonella typhimurium. PMID- 11400560 TI - A pseudoguaiane sesquiterpene xylopyranoside from Echinops hussoni. AB - A new pseudoguaiane sesquiterpene xylopyranoside ester, named echusoside (1), was isolated from the aerial parts of Echinops hussoni Boiss. (Asteraceae) collected from the southeastern border of Egypt. The structure elucidation of echusoside was based primarily on 1D, 2D-NMR analyses and chemical derivatization. This is the first report for a pseudoguainane sesquiterpene in the genus Echinops. PMID- 11400561 TI - Diterpenoid alkaloids from Delphinium virgatum poiret. AB - From the aerial parts of Delphinium virgatum Poiret, which is an endemic plant growing wildly in Turkey, a new norditerpenoid alkaloid, N-deethylperegrine alcohol, has been isolated along with the known alkaloids peregrine, peregrine alcohol, davisinol, hetisine, hetisinone and atisine. PMID- 11400562 TI - Triterpenoids and other compounds from Salvia roborowskii Maxim. AB - Two new triterpenoids, 2 alpha-hydroxy-3 beta-methoxyurs-12-en-28-oic acid and 3 alpha-hydroxy-2 alpha-methoxyurs-12-en-28-oic acid as well as 25 known compounds were isolated from the whole plant of Salvia roborowskii Maxim. Their structures were elucidated by means of spectral data and chemical transformation. This is the first report on the chemical constituents of this plant. The presence of eugenyl beta-D-glucopyranoside 22 in a plant of the genus Salvia is also reported for the first time. PMID- 11400563 TI - [Reaction of 2-4-aminobenzensulfonamide structures with thymol-sodium hypochlorite]. PMID- 11400564 TI - Capillary electrophoretic analysis of hydroxycinnamic acids from Ononis arvensis L. PMID- 11400565 TI - In vitro neuromuscular effects of droperidol in rats. PMID- 11400566 TI - Rhamnazin-4'-O-beta-[apiosyl(1-->2)] glucoside as a means of antioxidative defense against tetrachloromethane induced hepatotoxicity in rats. PMID- 11400567 TI - DNA-binding properties and cytotoxicity activity of novel aromatic amidines in cultured human skin fibroblasts. PMID- 11400568 TI - Effects of natural products containing acylresorcinol partial structures on cyclooxygenases and 5-lipoxygenase. PMID- 11400569 TI - Passive cigarette smoking induced changes in reactivity of the aorta in rabbits: effect of captopril. PMID- 11400570 TI - [Pathology of acute myocardial infarction]. PMID- 11400571 TI - [Acute heart infarct--new definition and diagnostic techniques]. PMID- 11400572 TI - [Acute heart infarct: epidemiology and pre-hospitalization phase]. PMID- 11400573 TI - [Thrombolytic therapy for acute myocardial infarct]. PMID- 11400574 TI - [Non-invasive acute therapy of cardiac infarction]. PMID- 11400575 TI - [Acute myocardial infarction: thrombolytic therapy in ambulance?]. PMID- 11400576 TI - [Interventional therapy of the acute cardiac infarct]. PMID- 11400577 TI - [Pharmacotherapy after cardiac infarction]. PMID- 11400578 TI - [Secondary prevention after cardiac infarct; therapeutic efficiency--cost-benefit ratio]. PMID- 11400579 TI - [Cutaneous paraneoplasia]. PMID- 11400580 TI - [Diffuse localized lesions of brain MRI in a 33 year old female with cognitive impairments]. PMID- 11400581 TI - [Thyreotoxic hypokalemic paralysis; presentation of symptoms in three case reports]. PMID- 11400582 TI - [Attending physician between therapeutic freedom and practice guidelines]. PMID- 11400583 TI - [Has there really been an exoneration of the breakfast egg?]. PMID- 11400584 TI - [Therapy with cellular blood components]. PMID- 11400585 TI - [Patent rights to human genes. 10th Meeting of the Board of Curators of Gastro Liga e.V. on December 14, 2000 in Munich]. PMID- 11400591 TI - [Repair and revision 4. Cracked tooth and crown fractures: diagnostics and treatment]. AB - Trauma to the teeth can result in the formation of cracks in the crown or root of the tooth. The location and the extent of these cracks determine the proper treatment choice. This treatment is oriented towards saving pulp vitality and/or tooth integrity. In this paper the diagnosis of these cracks is described as well as the desired treatment. These are discussed relative to the location of the cracks, namely in the clinical crown of the tooth, resulting in enamel infraction, cracked tooth and crown fractures, and in the root of the tooth, resulting in a vertical or horizontal root fracture. PMID- 11400592 TI - [Stability of orthodontic-maxillofacial surgical treatment of anterior open bite deformities]]. AB - A sample of 267 patients with maxillary hyperplasia, a Class I or Class II occlusion and anterior open bite, collected from three different institutions, was analysed regarding stability after Le Fort I intrusion osteotomies or bimaxillary osteotomies. Skeletal and dento-alveolar stability of the maxilla, postional changes of the mandible and of incisors were evaluated on cephalometric radiographs. The stability of maxillary arch dimensions after correction of the open bite is measured on dental casts. Patients with anterior open bite, treated with a Le Fort I osteotomy in one-piece or in multi-segments, with or without bilateral sagittal split osteotomy exhibited good skeletal stability of the maxilla. Rigid internal fixation showed better maxillary and mandibular stability than intraosseous wire fixation. Considerable relapse of transverse dimensions, however, was measured after orthodontic and surgical expansion. The mean overbite at the 69 months follow-up was 1.24 mm and lacking of overlap between opposing incisors was present in 19%. PMID- 11400593 TI - [How do Dutch dentists deal with patients' rights in practice?]. AB - In the Netherlands patients rights are regulated in the 'Medical Treatment Contract Act'. This article describes how dentists deal with those rights in daily practice. From the registers of the Dutch Dental Association 806 dental practitioners were drawn at random. They received a questionnaire on this topic; 41.6% responded. The results show that the implementation of patient rights in dental practice is far from ideal. Over a third of the respondents has had complaints about a lack of information from their patients. Respondents who attended post-graduate courses on dentist-patient communication dealt better with a number of patient rights than respondents who did not attend such courses. Confusion over how to interpret the rights of patients and the duties of the dentist could underlie the lack of implementation of those rights. PMID- 11400594 TI - [Cloning: applications in humans 2. Ethical considerations]. AB - Reproductive cloning in adults/children evokes unfavourable reactions. Direct objections are that cloning is unnatural, that it affects human dignity and violates the individual's right to genetic uniqueness. Consequential objections concern unjustified health risks for the progeny, unjustified psychosocial risks for the clone child and the risk of cloning for eugenetic purposes. There is consensus that reproductive cloning of existing persons is unjustify as yet because of the health risks for the offspring. Reproductive cloning of embryos is possible by means of nucleus transplantation and of embryo splitting. The ethical analysis of reproductive cloning of embryos depends on the purposes and applications. At least some of the moral objections against cloning of adults/children are not or not completely applicable to reproductive cloning of embryos. Conditions to be put to reproductive cloning of embryos are efficacy, safety and, at least for the time being, avoidance of asynchrony in transferring identical embryos. The ethical aspects of its application in the context of genetical reproductive techniques must be evaluated separately. Therapeutic cloning may be acceptable if alternatives are lacking. PMID- 11400595 TI - [Head-neck manifestations of syphilis]. PMID- 11400596 TI - [Bipolar disorder]. PMID- 11400597 TI - [Dental education via Internet]. PMID- 11400598 TI - [Is an alcohol-based mouthwash a risk factor for oral cancer?]. PMID- 11400599 TI - [Dr. F. Lobbezoo wins the Annual Prize 2000]. PMID- 11400600 TI - [What all our readers experienced. ... My most exciting emergency]. PMID- 11400601 TI - [Post-zoster neuralgia. Local acetylsalicylic acid stops pain for 5 hours]. PMID- 11400602 TI - [Elderly patients. Patients not fitting into a category are labeled as Alzheimer disease patients]. PMID- 11400603 TI - [New therapy for panic patients. Desensitization with computer? (interview by Dr. Beate Schumacher)]. PMID- 11400604 TI - [Coronary heart disease patients and sex. Death in the extramarital bed]. PMID- 11400605 TI - [Video images from the small intestine. A capsule delivers better results than currend endoscopy]. PMID- 11400606 TI - [Allergy research: news in epidemiology, diagnosis and therapy. What applies to general practice?]. AB - In the last few decades, the prevalence of allergic diseases has increased appreciably--for reasons that have not yet been identified. Currently research is concentrated on genetic basics, on the role of cytokines, T-helper and suppressor cells in the allergic sensitization process, and on the involvement of enzymes and chemokines in the triggering of an allergic reaction in the sensitized organism. New information provided by the results of research of relevance for the doctor's office concern the diagnosis of allergies, the avoidance of allergens, and prevention and immunotherapy. Today, improved methods of detecting IgE are available, and the atopy patch test has been shown to be superior to the prick and radio-allergo-sorbent tests. The measurement of allergens with strip tests in the domestic setting of the patient has improved the chances of avoiding allergen contact and prevention. Therapeutic innovations are to be found in particular in climatotherapy, the development of new antihistaminic agents as well as new allergen extracts and adjuvants for specific hyposensitization. In addition, initial results obtained with monoclonal IgE antibodies and an interleukin-4 receptor antagonist are very promising. PMID- 11400607 TI - [Allergy diagnosis for general practice. Proper testing!]. AB - The aim of allergy testing is to establish unequivocally the (possible) allergenic nature of the condition, and also to identify the triggering allergen(s). The diagnostic work-up of allergic diseases proceeds in accordance with a stepped schema. The individual steps comprise a history, physical examination, skin tests, in vitro diagnostic procedures and challenge (provocative) testing. With regard to skin tests, the major ones are the prick and scratch test, open and closed patch test and the intracutaneous test. The present article identifies the indications for the various tests, and describes how they are performed, and also discusses the usefulness of the individual in vitro methods and challenges. PMID- 11400608 TI - [Increase in practice failure. Are these colleagues to blame?]. PMID- 11400609 TI - [Insect venom allergy. Removing the sting of killer bees!]. AB - Systemic anaphylactic reactions to bee or wasp stings are potentially life threatening. Up to 5% of the population are affected. In the large majority of patients, an insect venom allergy can be diagnosed on the basis of the medical history, skin tests and determination of venom-specific serum IgE antibodies. Sensitization to insect venom (only "positive" test results) and insect venom allergy ("positive" test results and a history of systemic reactions of the immediate type to stings) must be carefully differentiated. Hyposensitization with bee or wasp venom reliably protects almost all patients with systemic sting reactions of the immediate type. Nevertheless, this unproblematic treatment of a potentially life-threatening allergy fails to reach many patients. If a sting challenge or a "field sting" is tolerated during the period of desensitization, treatment can be terminated after three to five years. If, however, the individual patient is at a particular risk, hyposensitization must be continued for a longer period, possibly even life-long. PMID- 11400610 TI - [Emergencies in general practice: medical autopsy. Don't become a homicide accomplice!]. PMID- 11400611 TI - [Hypertensive crisis, hypertensive emergency. Nitrate spray and ambulance--this can also be wrong]. PMID- 11400612 TI - [Morning fatigue in hypertensive snorers. When should obstructive sleep apnea syndrome be excluded?]. PMID- 11400613 TI - [Asthma patient with fever 6 months after a trip to Kenya. Can this still be a tropical disease?]. PMID- 11400614 TI - [67-year-old Ukrainian patient with cough since childhood. What is the cause of severe hemoptysis? Bronchiectasis]. PMID- 11400615 TI - [What are the options other than smoking cessation? Public health illness COPD calls in experts for the plan]. PMID- 11400616 TI - [Effective in more than asthma patients. Good reasons for inhaled steroids]. PMID- 11400617 TI - [Heartburn and dyspeptic symptoms. Almost all sour types respond to antacids]. PMID- 11400618 TI - [Chewable leukotriene antagonist. New therapy option for asthmatic patients of preschool age]. PMID- 11400619 TI - [Heart failure. With the AT1-blocker less frequent rehospitalization]. PMID- 11400620 TI - [Essential hypertension. What is the role of insulin?]. PMID- 11400621 TI - [Forensic physicians complain about invalid death certificates. Carelessness during the last consultation? (interview by Dr. Beate Schumacher)]. PMID- 11400622 TI - [Current recommendations for air travel fitness. "Doctor, may I fly?"]. PMID- 11400623 TI - ["Is there a physician on board?" The hit list of medical emergencies on board airplanes]. PMID- 11400624 TI - [First aid on board passenger airplanes. What is available?]. PMID- 11400625 TI - [Emergencies in general practice: bacterial meningitis. The general practitioner as life saver]. PMID- 11400626 TI - [Exotic pollen now also in Europe. Allergy tests negative, runny nose despite tests]. PMID- 11400627 TI - [Normotensive in general practice, hypertensive at work. Undetected infarct candidates]. PMID- 11400628 TI - [The wound. 2: General principles for treatment]. PMID- 11400629 TI - [Diagnostic quiz. Epigastric pain in unremarkable ultrasound imaging. Cystic duct calculus]. PMID- 11400630 TI - [Precise direction or flexible self-control. How strictly must the asthma patient adhere to medical advice?]. PMID- 11400631 TI - [Therapy of hypertension in general practice. When pressure must be lowered quickly]. PMID- 11400632 TI - [Even the best motivated patient forgets his medicines. What is your concept to control noncompliance?]. PMID- 11400633 TI - [70-year-old patient with isolated systole hypertension. Patient should certainly be treated]. PMID- 11400634 TI - Programming for adolescent health and development. Report of a WHO/UNFPA/UNICEF Study Group on Programming for Adolescent Health. AB - One in every five persons is an adolescent (10-19 years of age). What happens, or does not happen, during the second decade of life has implications that last throughout a lifetime and affect both individual and public health. What sets adolescents apart from children is the increasing autonomy they demonstrate. Their own decisions, behaviours and relationships increasingly determine their health and development. Moreover, adolescence brings with it expanded capacities- for abstract thought and contemplating the future, for empathy and idealism, for critical thought including the questioning of self and others, and for reproduction. Yet the use of these new capacities is dependent on the environment in which adolescents live. So while adolescents display more self-reliance than children, they lack the status and resources of adults. Indeed, they are often dependent on adults to meet many of their basic needs. The consolidation of knowledge and experience acquired through the WHO/UNFPA/UNICEF Study Group on Programming for Adolescent Health is presented in this report. Verified by research, the report describes the guiding concepts and major interventions that are necessary components for country programming for adolescent health and development. The report asserts the value of addressing the health and development of adolescents simultaneously. It indicates the emerging evidence that actions to meet adolescents' needs for development also discourage them from adopting high-risk behaviours and protect them from the situations that lead to the major health problems. The report is illustrated by examples of programming efforts from around the world. PMID- 11400635 TI - Marine macroalgae in polar regions as natural sources for volatile organohalogens. AB - Marine macroalgae species from the polar regions were investigated for their importance as natural sources of volatile halogenated compounds released into the biosphere. Several different halogenated C1 to C4 hydrocarbons were identified and their release rates determined. The compounds contained mainly bromine and iodine, and form was the dominant compound released. Although an annual atmospheric input of approximately 10(8)-10(10) g bromine and 10(7)-10(8) g iodine was calculated from the release rates, marine macroalgae are apparently not the major source on a global scale, as the release is up to four orders of magnitude lower than a presumed annual flow from the oceans. Despite this, macroalgae may be more important on a local scale due to their occurrence at a high biomass in the coastal regions. The present paper gives an overview about studies done on the release of volatile halocarbons by macroalgae from polar regions. Furthermore, the function of these compounds in the macroalgal metabolism is discussed. PMID- 11400636 TI - Pesticide residues in products of plant origin in the European Union. Sampling strategy and results from the co-ordinated EU monitoring programmes in 1996 and 1997. AB - The results of a co-ordinated monitoring programme for pesticide residues in the European Union and Norway carried out in 1996 and 1997 are presented. The aim of this programme is to work towards a system, which makes it possible to estimate actual dietary pesticide intake for the population of the European Union. Based on a statistically derived sampling plan and within the limited number of pesticides/commodities analysed, the most critical pesticides (benomyl group and dithiocarbamates) and commodities (mandarine and lettuce) were identified. In case of detected non-compliances, repeated sampling and, if necessary, enforcement actions are to be taken by national authorities. The programme will be continued in the next years. PMID- 11400637 TI - Environmental risks of chemicals and genetically modified organisms: a comparison. Part I: Classification and characterisation of risks posed by chemicals and GMOs. AB - Risks can be characterised by several parameters. A risk is commonly defined to be the product of the extent of damage and the probability of its occurrence. But there are several other characteristics to be taken into account: degree of certainty in determining extent and probability, persistency, ubiquity, irreversibility, delay effect and mobilisation potential. As potential risks of genetically modified plants (GMPs), resistance to antibiotics, impact on non target organisms, spread of genes and GMOs, and secondary consequences, e.g. on cultivation practice, are discussed in detail. Risks of GMPs are, in general, characterised by high uncertainty of the magnitude and probability of damage, a high mobilisation potential and a delay effect. PMID- 11400638 TI - Using sustainable development indicators as a supplementary measure for the integrated management of environmental information system in Taiwan. AB - Environmental Information System (EIS) transfers environmental data and information to audiences in any field of knowledge or activity interested in environmental problems. Currently, numerous conventional EISs or Environmental Databases (EDs) are available in Taiwan. However, they fail to adequately address the strong correlations among the environmental, social and economic components. Notably, Sustainable Development Indicators (SDIs) combine environmental, social and economic dimensions, as well as illustrate the links among systems. Subsequently, developing a set of SDIs will provide an excellent opportunity to systematically consolidate the information scattered among many archives, as well as promoting an integrated EIS. This work presents an experimental model referred to as the 'Sustainable Development Information System of the Island Taiwan' (SDISIT). This system was developed to assist the integrated management of either EIS or ED. The Factor Analysis Method (FAM) is one tool that may prove useful in easing the developing process. Furthermore, by examining the correlations between the indicators, this method can apply existing SDIs to identify the structure of interrelationships among indicators. PMID- 11400639 TI - Environmental chemistry and environmental engineering education in Hungary. PMID- 11400640 TI - Incineration and human health. State of knowledge of the impacts of waste incinerators on human health. AB - After pollutants from an incineration facility disperse into the air, some people close to the facility may be exposed directly through inhalation or indirectly through consumption of food or water contaminated by deposition of the pollutants from air to soil, vegetation, and water. For metals and other pollutants that are very persistent in the environment, the potential effects may extend well beyond the area close to the incinerator. Persistent pollutants can be carried long distances from their emission sources, go through various chemical and physical transformations, and pass numerous times through soil, water, or food. PMID- 11400641 TI - Bovine spongiform encephalopathy (BSE)/transmissible spongiform encephalopathy/mad cow disease. PMID- 11400642 TI - Dioxin contamination in food. Bayreuth, Germany, from September 28 to October 1, 2000. AB - Dioxin and PCB monitoring programs for food and feeding stuff in most countries of the world, including many European Countries are currently inadequate. Better control of food production lines and food processing procedures is needed to minimize entry of dioxin to the food chain and will help to avoid dioxin contamination accidents. This would also improve the ability to trace back a possible contamination to its source. European guidelines for monitoring programs should be established to ensure comparable and meaningful results. These guidelines should define the minimum requirements for the design of monitoring programs, analytical methods, and quality assurance. Though data from Northern Europe shows that the general population exposure to dioxin and PCB has decreased during the last ten years these compounds continue to be a risk of accidental contamination of the food chain. The most prominent recent example is the Belgian dioxin contamination of feeding stuff in 1999. The Belgian dioxin contamination was not detected due to dioxin monitoring programs but by their direct biological effects seen in animals. Four other cases of dioxin contamination have been detected in Europe since 1997 due to local monitoring programs. One of them (citrus pulp pellets 1998) was in a much larger scale than the Belgian dioxin contamination. The general population's exposure to dioxins and PCBs is still in the same range (1-4 pg WHO-TEQ/kg body weight and day) as the recently revised WHO tolerable daily intake (TDI). There is concern that short-term high level exposure to dioxins, furans, and PCB may cause biological effects on the human fetal development and further research is required. Further actions to control sources building on considerable advances already made in many countries may need to be supplemented by measures to prevent direct contamination of feeding stuff or food to reduce general population exposure further. PMID- 11400643 TI - Tracking of phenol degrading genotype. AB - A simple and rapid protocol has been developed for monitoring of the phenol degrading population in the environmental samples. PCR protocol uses the total DNA prepared from the samples as a template in the amplification reaction. Primers have been designed from the sequence data for Dmp and Phe operon encoding multi-component and single component phenol oxidizing system, respectively. The phenol degrading genotype in various samples have been demonstrated using the developed PCR protocol. PMID- 11400644 TI - Flow analysis of PCDD/Fs for Tarragona Province, Spain. A preliminary inventory. AB - The present study was intended to establish an inventory of PCDD/F emissions in Tarragona Province (Catalonia, NE Spain), as a preliminary phase in the development of a flow analysis of PCDD/Fs in this Province. In 1999, global PCDD/F emissions reached a value of 2.24 g I-TEQ/y, which means a density of 3.8 micrograms I-TEQ/inhabitant/y. The low amount of PCDD/Fs emitted to air by the only municipal solid waste incinerator in the Province (approximately 0.04% of the total) has been one of the most notable results. As a reflection of uncertainties in the estimates for individual sources, the 90th percentiles of PCDD/F releases for 1999 ranged up to 4.1 g I-TEQ/y. PMID- 11400645 TI - Forest ecosystems and the changing patterns of nitrogen input and acid deposition today and in the future based on a scenario. AB - A global assessment of the impact of the anthropogenic perturbation of the nitrogen and sulfur cycles on forest ecosystems is carried out for both the present-day [1980-1990] and for a projection into the future [2040-2050] under a scenario of economic development which represents a medium path of development according to expert guess [IPCC IS92a]. Results show that forest soils will receive considerably increasing loads of nitrogen and acid deposition and that deposition patterns are likely to change. The regions which are most prone to depletion of soils buffering capacity and supercritical nitrogen deposition are identified in the subtropical and tropical regions of South America and Southeast Asia apart from the well known 'hotspots' North-Eastern America and Central Europe. The forest areas likely to meet these two risks are still a minor fraction of the global forest ecosystems, though. But the bias between eutrophication and acidification will become greater and an enhanced growth triggered by the fertilizing effects of increasing nitrogen input cannot be balanced by the forest soils nutrient pools. Results show increasing loads into forest ecosystems which are likely to account for 46% higher acid loads and 36% higher nitrogen loads in relation to the 1980-1990 situation. Global background deposition of up to 5 kg N ha-1 a-1 will be exceeded at more than 25% of global forest ecosystems and at more than 50% of forest ecosystems on acid sensitive soils. More than 33% of forest ecosystems on acid sensitive soils will receive acid loads which exceeds their buffering capacity. About 25% of forest areas with exceeded acid loads will receive critical nitrogen loads. PMID- 11400646 TI - Thrombotic risk in children with acute lymphoblastic leukemia. PMID- 11400647 TI - Unrelated cord blood transplant experience by the pediatric blood and marrow transplant consortium. AB - Cord blood (CB) has emerged as a potential source of hematopoietic stem cells for patients who are in need of hematopoietic stem cell transplant (HSCT). The authors analyzed the Pediatric Blood and Marrow Transplant Consortium's (PBMTC) data of consecutive unrelated CB transplants performed during the initial 2 years of using placental blood grafts. From January 1995 to December 1996 PBMTC performed a total of 44 unrelated CB transplant for a variety of diseases consisting of acute leukemias (n = 29), congenital conditions (n = 9), and bone marrow failure (n = 6). There were 15 females and 29 males with median age of 5 years (range 0.4-20.6 years) and median weight of 18.2 kg (range 6.3-70 kg). The median volume of CB units was 80 mL (range 44.5-215 mL) and the median cell dose given was 4.3 x 10(7)/kg of recipient weight (range 1.1-23 x 10(7)/kg). Techniques used for human leukocyte antigen (HLA) matching were serologic typing for class I HLA antigens and high-resolution molecular typing for HLA-DRB1 alleles. HLA disparities were as follows: 4 were 6/6 matches, 21 were 5/6, 15 were 4/6, and 4 were 3/6. Twenty-nine (66%) of CB units were DRB1 matched with recipients. Conditioning regimens consisted of either total body irradiation containing (n = 31) or chemotherapy only (n = 11) regimens. All but 3 patients receive cyclosporine as part of graft vs. host disease (GvHD) prophylaxis in combination with either methotrexate (MTX) or methylprednisolone (Pred). The other 3 patients had FK506 and MTX for GvHD prophylaxis. Myeloid engraftment (absolute neutrophil count > or = 500) occurred at a median of 21 days (range 10 43 days) and platelet > or = 50,000/mm3 was noted at a median of 44 days (range 16-102 days). Eight patients died too early (< day + 28) for evaluation of engraftment (5 for infection, 2 for multiorgan failure, 1 for toxic epidermolysis). The probability of having grade II-IV acute GvHD for all patients was 44 +/- 0.7%. The incidence of a GvHD is similar for 4/6 and 5/6 antigen when DRB1 matched, at 47 and 52%, respectively. Chronic GvHD was noted in 28% of patients surviving > 90 days. The Kaplan-Meier estimate of 4-year event-free survival was 43%. A Cox model for analysis of factors associated with survival was DRB1 matching, p = .001; cell dose, p = .009; and younger age, p = .03. In conclusion, CB transplant offers a good alternative to bone marrow transplant Although GvHD occurs, it is usually of low severity despite the high frequency of multiple HLA antigen mismatches. It also appears that a 4/6 is as good as a 5/6 matched antigen CB unit when DRB1 matched especially in the pediatric setting. PMID- 11400648 TI - Successful pregnancy after high-dose cyclophosphamide and ifosfamide treatment in two postpubertal women. AB - Alkylating agents, especially cyclophosphamide, are known to have a destructive effect on the ovaries and to result in sterility in many young women treated with these drugs. This is especially true when the treatment is given to postpubertal women. The authors describe 2 postpubertal women aged 16 and 25 suffering from Ewing sarcoma who were treated with the very aggressive Sloan-Kettering protocol, which includes high-dose cyclophosphamide and ifosfamide in addition to other drugs. Both women had spontaneous pregnancies and delivered normal babies. The significance of these cases in view of the experimental various reproductive preservation measures offered to such women is discussed. PMID- 11400649 TI - Prolonged second response to cisplatin, etoposide, and ifosfamide in a child with a recurrent brainstem glioblastoma. AB - The prognosis for patients with malignant brainstem tumors is poor. The authors report on a 6-year-old girl with a biopsy proven pontine glioblastoma, who, after initial chemo-radiotherapy and tumor progression, experienced a prolonged second response to a salvage therapy consisting of cisplatin, etoposide, and ifosfamide. The patient recovered from her life-threatening condition with almost complete resolution of all neurologic deficits paralleled by a dramatic shrinkage of the tumor documented by magnetic resonance imaging. PMID- 11400650 TI - Pleuropulmonary blastoma: an aggressive intrathoracic neoplasm of childhood. AB - A case of a 4-year-old girl with pleuropulmonary blastoma is reported. Surgical resection of the tumor was performed and histologic examination revealed pleuropulmonary blastoma with rhabdomyosarcomatous differentiation. Postoperative chemotherapy was administered and 3 weeks after initiation of treatment protocol a second site of lesion in the retroperitoneum was revealed with extension to the mediastinum, which shared similar mesenchymal neoplastic characteristics to the previously diagnosed primary lesion. The girl died 4 1/2 months after initial evidence of disease because of brain metastasis, indicating a very aggressive neoplasm unresponsive to treatment. PMID- 11400651 TI - Childhood T-cell acute lymphoblastic leukemia with four distinct immunophenotypes representing different stages of T-cell development. AB - The authors report on a 14-year-old boy who developed T-cell acute lymphoblastic leukemia (FAB:L1) displaying 4 immunophenotypically distinct leukemic cell populations by 3-color immunofluorescence staining. Cytogenetic analysis at diagnosis showed 46,XY,add(4)(p16)[12]/46,XY[2]. A single rearrangement of the T cell antigen receptor beta- and gamma-chain genes in these cells indicated monoclonality of the leukemic cells. These findings suggest that leukemic blast cells of monoclonal origin in this case were divided into 4 immunophenotypic populations, representing various stages of differentiation. PMID- 11400652 TI - Fibrolamellar hepatocellular carcinoma with skeletal metastases. AB - Skeletal metastases is relatively rare in hepatocellular carcinoma and accounts for 4-16% of extrahepatic metastases. The authors report a 13-year-old girl with fibrolamellar hepatocellular carcinoma, who rejected further systemic chemotherapy following hepatic lobectomy and experienced sternal and vertebral painful metastases nearly 5 years after the operation. The sternal metastatic lesion was removed surgically, whereas external irradiation was delivered to the lumbar vertebral lesion. The patient received no systemic treatment following metastases and died with widespread disease. Despite metastatic disease, the patient survived 6.5 years following the initial diagnosis. PMID- 11400653 TI - Successful treatment of neonatal rhesus hemolytic anemia with high doses of recombinant human erythropoietin. AB - The authors report the use of high-dose recombinant erythropoietin (r-HuEPO) in a full-term newborn baby with severe postnatal rhesus hemolytic anemia (RHA). Hemoglobin (Hb) value and reticulocyte count at day 13 of life were 59 g/L and 234 x 10(9)/L, respectively. Three days after the r-HuEPO (870 U/kg/d) administration, reticulocyte count had increased more than 4-fold and Hb rose to 73 g/L. r-HuEPO was gradually decreased after 18 days of treatment. No major side effect was observed. In selected cases of severe anemia due to hemolytic disorders, transfusions may be avoided by the use of high doses of r-HuEPO. PMID- 11400654 TI - Unusual presentation of mastoid eosinophilic granuloma in a young patient. AB - Eosinophilic granuloma is a well-recognized form of Langerhans cell histiocytosis, most commonly involving the skull bones, usually with an excellent prognosis. Recurrent and difficult to recognize osteolytic lesions of the skull are encountered only rarely. A patient with recurrent eosinophilic granuloma of the skull is reported. In spite of appropriate multimodality treatment, there were several recurrences, most recently with involvement of the mastoid process. Imaging studies revealed extensive involvement of surrounding structures with expansion of the tumor into the middle cranial fossa and slight pressure on the antero-medial portion of the temporal lobe of the brain. Despite extensive involvement, the patient had no complaints. Because of the rarity of such silent and unpredictable lesions, a systematic approach with regular CT and MRI follow up is suggested. PMID- 11400655 TI - Etoposide as the basic and interferon-alpha as the maintenance therapy for Langerhans cell histiocytosis: a RTC. AB - The treatment of patients who suffer from a disseminated form of Langerhans cell histiocytosis (LCH) is still controversial. So far, few larger randomized studies have been performed. The authors present 3 patients with a disseminated form of LCH--4 months, 9 months, and 2 years old, respectively. The lesional Langerhans cells in each patient showed positive immunohistochemical reaction to S-100 protein and the presence of Birbeck granules was confirmed by electron microscopy. All the patients were treated with etoposide (VP-16), 200 mg/m2 for 3 consecutive days, with 15 cycles at intervals of 3 weeks between each cycle, followed by maintenance therapy with IFN-alpha. All 3 patients reached complete stabile remission. The patients were young, at high risk, with multiple-organ involvement of LCH, and two of them had obvious signs of organ dysfunction at presentation, suggesting a poor prognosis. All remain disease-free several years after therapy. The results suggest that INF-alpha may prevent recurrences in high risk patients. PMID- 11400656 TI - Detection of isolated distant metastasis in soft tissue sarcoma by fluorodeoxyglucose positron emission tomography: case report. AB - Fluorodeoxyglucose (FDG), labeled with F-18, is a glucose analog that accumulates in cells in proportion to the rate of glucose metabolism, and increased carbohydrate metabolism has been recognized as a feature of malignant cells versus normal cells. In addition, it permits the detection of metastases not discovered by bone scan. Although detection of the primary site of disease is usually accomplished well with conventional techniques, the performance of FDG positron emission tomography (PET) may be useful to determine metastases that are not clinically evident. The authors describe a case of early detection of distant metastases by FDG-PET in a young patient diagnosed with rhabdomyosarcoma of the hand. PMID- 11400657 TI - A convenient method of granulocyte transfusion using a hydroxylethyl starch (HESPAN) containing a bag. PMID- 11400658 TI - Pseudohyperkalemia due to reactive thrombocytosis in an infant with yolk sac tumor. PMID- 11400659 TI - [AIDS--a disease we must be prepared to treat]. PMID- 11400660 TI - [Protein design of HIV-1 infection inhibitors]. PMID- 11400661 TI - [A tool for antibiotic resistance detection]. PMID- 11400662 TI - [Hope for a vaccine against ebola virus]. PMID- 11400663 TI - [HIV integrase inhibitors: a promising development in AIDS research]. PMID- 11400664 TI - [New weapons against multiresistant bacteria?]. PMID- 11400665 TI - [How did it all begin? Discovery and origin of HIV]. PMID- 11400666 TI - [Molecular modeling in the battle against AIDS. Drugs design in the development of substrate-like HIV protease inhibitors]. PMID- 11400667 TI - [Only when we know the enemy, can we correctly fight him. The structure of the HIV]. PMID- 11400668 TI - [An important point of attach in AIDS treatment. HIV protease inhibitors: state of the art]. PMID- 11400669 TI - [Alternatives to peptidomimetics. New molecular classes as HIV protease inhibitors?]. PMID- 11400670 TI - [Good, but adverse effect-producing drugs. The importance of HIV protease inhibitors in combination therapy]. PMID- 11400671 TI - [What should we pay attention to in combination treatment of HIV infected patients? Interaction problems of protease inhibitors]. PMID- 11400672 TI - [Characterization of resistance in HIV. Comparison of geno- and phenotypic resistance to HIV-1 protease inhibitors]. PMID- 11400673 TI - [Formulation of HIV protease inhibitors. Pharmaceutical technology and biopharmaceutic aspects of HIV protease inhibitors]. PMID- 11400674 TI - [How should pharmacists assist in combination therapy? Pharmaceutical care of HIV patients]. PMID- 11400675 TI - [Just expensive or also cost effective? Pharmacoeconomic aspects of HIV therapy]. PMID- 11400676 TI - [Parkinson's disease: diagnostic and therapeutic criteria. Consensus conference proceedings. Paris, France, 3 March 2000]. PMID- 11400677 TI - Proceedings of the 6th Equine Colic Research Symposium. Georgia, November 1998. PMID- 11400678 TI - Proceedings of the 2nd International Conference on Scanning Probe Microscopy, Sensors and Nanostructures. Heidelberg, Germany, May 28-31, 2000. PMID- 11400679 TI - Dealing with malaria in the last 60 years: aims, methods and results. Proceedings of a conference. May 11-14, 1998. Sleepy Hollow, New York, USA. PMID- 11400680 TI - 20th Workshop of the Study Group on Artificial Insulin Delivery, Pancreas and Islet Transplantation (AIDPIT) of the European Association for the Study of Diabetes (EASD). Igls, Austria, January 28-30, 2001. Abstracts. PMID- 11400681 TI - The Ovary: Genesis, Function, and Failure. Proceedings of a workshop. Bethesda, Maryland, USA. March 30-31, 2000. PMID- 11400682 TI - Altered expression of estrogen receptor-alpha variant messenger RNAs between adjacent normal breast and breast tumor tissues. AB - Using semiquantitative reverse transcription-polymerase chain reaction assays, we investigated the expression of variant messenger RNAs relative to wild-type estrogen receptor (ER)-alpha messenger RNA in normal breast tissues and their adjacent matched breast tumor tissues. Higher ER variant truncated after sequences encoding exon 2 of the wild-type ER-alpha (ERC-4) messenger RNA and a lower exon 3 deleted er-alpha variant (ERD3) messenger RNA relative expression in the tumor compartment were observed in the ER-positive/PR-positive and the ER positive subsets, respectively. A significantly higher relative expression of exon 5 deleted ER-alpha variant (ERD5) messenger RNA was observed in tumor components overall. These data demonstrate that changes in the relative expression of ER-alpha variant messenger RNAs occur between adjacent normal and neoplastic breast tissues. We suggest that these changes might be involved in the mechanisms that underlie breast carcinogenesis. PMID- 11400683 TI - Essays celebrating the centenary of the pioneer neuroembryologist, Viktor Hamburger. PMID- 11400684 TI - [Two surgical cases of pulmonary aspergilloma with different postoperative prognosis]. PMID- 11400685 TI - [Case of pulmonary aspergillosis with severe hemoptysis successfully treated by right upper lobectomy following TAE]. PMID- 11400686 TI - [Case of pulmonary aspergillosis following neurosurgery treated by pneumonectomy]. PMID- 11400687 TI - [Two surgical cases of pulmonary aspergilloma]. PMID- 11400688 TI - [Surgical treatment of patients with pulmonary aspergilloma]. PMID- 11400690 TI - Annual Conference of Engineering and the Physical Sciences in Medicine. 5-9 November 2000. Newcastle, Australia. Abstracts. PMID- 11400689 TI - [Present status of studies on pulmonary aspergillosis--special reference to the pathogenesis and progression]. PMID- 11400691 TI - [Proceedings of the 7th National Congress "Pollens, Particles, Pollution". April 2000, Strasbourg, France]. PMID- 11400692 TI - The biochemical basis of anthelmintic action and resistance. AB - The most commonly used modern anthelmintics include the benzimidazoles, the nicotinic agonists. praziquantel, triclabendazole and the macrocyclic lactones. These drugs interfere with target sites that are either unique to the parasite or differ in their structural features from those of the homologous counterpart present in the vertebrate host. The benzimidazoles exert their effect by binding selectively and with high affinity to the beta-subunit of helminth microtubule protein. The target site of the nicotinic agonists (e.g. levamisole, tetrahydropyrimidines) is a pharmacologically distinct nicotinic acetylcholine receptor channel in nematodes. The macrocyclic lactones (e.g. ivermectin, moxidectin) act as agonists of a family of invertebrate-specific inhibitory chloride channels that are activated by glutamic acid. The primary mode of action of other important anthelmintics (e.g. praziquantel, triclabendazole) is unknown. Anthelmintic resistance is wide-spread and a serious threat to effective control of helminth infections, especially in the veterinary area. The biochemical and genetic mechanisms underlying anthelmintic resistance are not well understood, but appear to be complex and vary among different helminth species and even isolates. The major mechanisms helminths use to acquire drug resistance appear to be through receptor loss or decrease of the target site affinity for the drug. Knowledge on the mechanisms of drug action and resistance may be exploitable for the development of new drugs and may provide information on ways to overcome parasite resistance, respectively. PMID- 11400693 TI - Aquagenic pruritus relieved by tight fitting clothing. PMID- 11400694 TI - Persistent head lice following multiple treatments. PMID- 11400697 TI - Judging complementary and alternative medicine. PMID- 11400696 TI - Modern management of cancer-related intestinal obstruction. AB - Malignant-associated bowel obstruction remains a common and perplexing problem for patients with advanced gynecologic and gastrointestinal malignancies. The ability to locate and define its cause preoperatively has improved with the advent of computed tomography. Initial clinical experience with half-Fourier acquisition single-shot turbo spin-echo magnetic resonance imaging (HASTE MRI) and virtual colonoscopy is exciting. The surgical approach for primary obstructing colon cancer has become more aggressive, with experienced surgical groups doing one-stage procedures. Yet to be defined are guidelines for surgical management of obstructions occurring in the face of recurrent disease. Stent placement for upper and lower bowel obstructions is an option in nonoperable patients. Pharmacologic symptom management for intestinal obstructions consists of an opioid, an anticholinergic, and an antiemetic. Octreotide, either alone or added to the original regimen, will palliate symptoms that are resistant to the three-drug combination. PMID- 11400698 TI - An indigenous perspective on "Medicine and story". PMID- 11400699 TI - Breast samples are not representative of the breast as a whole. PMID- 11400700 TI - Sharing Communication. Proceedings of a conference. Birmingham, United Kingdom, 17-19 April 2001. PMID- 11400701 TI - Stimulated cord blood lymphocytes have a low percentage of Th1 and Th2 cytokine secreting T cells although their activation is similar to adult controls. PMID- 11400702 TI - Proceedings of the VIII International Symposium on Cardiac Bioprostheses. Cancun, Mexico, November 3-5, 2000. PMID- 11400703 TI - Historical image. Doctor Jacques Daviel. PMID- 11400704 TI - Historical image. Care of the handicapped. Guide dogs for the blind. PMID- 11400706 TI - Recurrent Epstein-Barr virus associated disease in a cardiac transplant patient: evolution from plasmacytic hyperplasia to diffuse large cell lymphoma. AB - An Epstein-Barr virus (EBV)-seronegative 31-year-old male underwent cardiac transplantation in 1991 for congenital cardiomyopathy. He presented with a protracted course of waxing and waning lymphadenopathy beginning four years after transplantation with eventual progression to a fulminant EBV-positive large cell lymphoma eight years after transplantation. Risk factors for the development of post-transplant lymphoproliferative disease in this patient, the importance of a standardized approach to pathology in assessing therapeutic options, and the management strategies used are discussed. PMID- 11400705 TI - Recurrent hernia of Petit's triangle: a case report. AB - The Authors describe a case of recurrent hernia of Petit's triangle in a 43-year old man. The most salient feature of lumbar hernias is the difficulty of their treatment. Numerous, more or less complex techniques have been proposed for the repair of lumbar hernias. The classic procedure is that of Dowd (1907). Repair in this case consists of a flap of fascia and aponeurosis from the gluteus medius muscle which is elevated and rotated to cover the defect. Many modifications of this technique have been proposed. The modern prosthetics mesh materials allow a tension-free repair of lumbar hernias without distortion of the normal anatomy. Correct placement of a prosthetic mesh is relatively simple. Recurrences are due not to the material used, but to patches which are too small or too tight. PMID- 11400707 TI - Early diagnosis of cystic fibrosis. PMID- 11400708 TI - Nursing home residence: quality of life among individuals with spinal cord injury. AB - OBJECTIVE: To examine the quality of life among individuals with spinal cord injury currently living in a nursing home vs. community residence. DESIGN: A case control design matched nursing home and community residents on numerous demographic and medical characteristics. Two groups of 37 individuals (nursing home vs. community residents) were matched case-for-case on age (i.e., within 10 yr), education, gender, race, marital status, and impairment level. Etiology of spinal cord injury, American Spinal Injury Association Impairment Index at the time of rehabilitation discharge, and injury duration, although not specifically matched, were not significantly different between groups (P > 0.05). Outcome measures included the Satisfaction With Life Scale, and the Craig Handicap Assessment and Reporting Technique (CHART). RESULTS: Results indicated that overall self-reported quality of life was significantly lower among nursing home residents compared with matched community-dwelling residents (mean = 18 and 14, respectively). Similarly, self-reported handicap was significantly higher among the nursing home residents compared with community residents (mean = 337 and 181, respectively), particularly in the areas of physical independence, mobility, occupational functioning, and economic self-sufficiency (P < 0.05). In contrast, the score on the CHART social integration subscale was similar across groups (P > 0.05). CONCLUSION: Despite matching numerous variables, the nursing home residents demonstrated significantly lower quality of life across multiple domains as compared with their community-dwelling counterparts. Additional research is needed to examine the specific mechanisms associated with these differences. PMID- 11400709 TI - Evaluating panic-specific factors in the relationship between suicide and panic disorder. AB - Work during the past decade has suggested an association between panic disorder and suicide (i.e., suicidal ideation and suicide attempts) that cannot simply be accounted for by co-occurring depression symptoms. To clarify the linkage between panic disorder and suicide, the association between panic-specific clinical and cognitive variables and suicide indicators were evaluated in patients with panic disorder (N=146). Analyses predicting the presence of suicidal ideation (positive, negative) after covarying the effects of a current mood disorder diagnosis and depression symptoms indicated a number of significant predictors including: (1) overall anxiety symptoms; (2) level of anticipatory anxiety; (3) avoidance of bodily sensations; (4) attentional vigilance toward bodily perturbations; and (5) phrenophobia (i.e., fear of cognitive incapacitation). Anxiety-specific variables did not account for unique variance in predicting prior history of suicide attempts. PMID- 11400710 TI - Effects of varying levels of anxiety within social situations: relationship to memory perspective and attributions in social phobia. AB - Cognitive-behavioral theorists (Clark & Wells, 1995: Clark, D. M. & Wells, A. (1995). A cognitive model of social phobia. In R. G. Heimberg, M. R. Liebowitz, D. A. Hope, & F. R. Schneier (Eds.), Social phobia: Diagnosis, assessment, and treatment (pp. 69-93). New York: Guilford Press; Rapee & Heimberg, 1997: Rapee, R. M., & Heimberg, R. G. (1997). A cognitive-behavioral model of anxiety in social phobia. Behaviour Research and Therapy, 35, 741-756.) propose that individuals with social phobia form mental images of themselves as if from an external point of view. Research by Wells and colleagues has shown that, when recalling anxiety-provoking social situations, individuals with social phobia are more likely to take an observer perspective (seeing oneself as if from an external point of view) whereas control subjects are more likely to take a field perspective (as if looking out through one's own eyes). Furthermore, this pattern is specific to social events, as both groups recall non-social events from a field perspective (see Wells, Clark & Ahmad, 1998: Wells, A., Clark, D. M., & Ahmad, S. (1998). How do I look with my minds eye: perspective taking in social phobic imagery. Behaviour Research and Therapy, 36, 631-634; Wells & Papageorigou, 1999: Wells, A. & Papageorgiou, C. (1999). The observer perspective: Biased imagery in social phobia, agoraphobia, and blood/injury phobia. Behaviour Research and Therapy, 37, 653-658). In the current study, individuals with social phobia took more of an observer perspective than non anxious controls when recalling high anxiety social situations. However, both groups took a predominantly field perspective for memories of medium or low anxiety social situations. As memory perspective has also been shown to be related to causal attributions, we examined this relationship in our sample. Memories of low, medium, and high anxiety social situations were differentially related to attributions for each group. Patients' attributions for their performance became more internal, stable, and global as the anxiety level of the situation increased, while the attributions of control subjects showed the opposite pattern. PMID- 11400711 TI - Evaluation of a cognitive-behavioural program for the management of chronic tic and habit disorders. AB - The aim was to evaluate the efficacy of a manualized cognitive-behavioural program based on habit reversal for the management of chronic tic disorder (CTD) and habit disorder (HD). Forty-seven CTD and 43 HD received a 4-month treatment program. Thirty-eight (22 CTD, 16 HD) were placed on a waitlist control group, which subsequently received treatment. The treatment approach combined awareness training, relaxation (including modification of a tension-producing style of action), and habit-reversal training, with more general cognitive restructuring of anticipations linked to ticcing. Sixty-five percent of completers reported between 75 and 100% control over the tic. At 2-year follow-up, 52% rated 75-100% control. There were also significant changes post-treatment in measures of self esteem, anxiety, depression and style of planning action. Successful tic/habit modification was associated in CTD and HD groups with successful change in style of planning action. There were no consistent differences in any outcome measures between CTD and HD groups. PMID- 11400712 TI - Anxiety sensitivity in the prediction of pain-related fear and anxiety in a heterogeneous chronic pain population. AB - The present study evaluated anxiety sensitivity, along with depression and pain severity, as predictors of pain-related fear and anxiety in a heterogeneous chronic pain population (n=68). The results indicated that the global anxiety sensitivity factor, as indexed by the Anxiety Sensitivity Index (ASI: Reiss, Peterson, Gursky & McNally, 1986: Reiss, S., Peterson, R. A., Gursky, M. & McNally, R. J. (1986). Anxiety, sensitivity, anxiety frequency, and the prediction of fearfulness. Behaviour Research and Therapy, 24, 1-8) total score, was a better predictor of fear of and anxiety about pain relative to the other relevant variables. Additionally, the physical concerns subscale of the ASI was a better predictor of pain-related fear dimensions characterized by high degrees of physiological symptoms and behavioral activation on both the Fear of Pain Questionnaire-III (FPQ-III; McNeil & Rainwater, 1998: McNeil, D. W. & Rainwater, A. J. (1998). Development of the Fear of Pain Questionnaire-III. Journal of Behavioral Medicine.) and Pain Anxiety Symptoms Scale (PASS; McCracken, Zayfert & Gross, 1992: McCracken, L. M., Zayfert, C. & Gross, R. T. (1992). The Pain Anxiety Symptoms Scale: Development and validation of a scale to measure fear of pain. Pain, 50, 67-73). In a related way, the ASI psychological concerns subscale was a better predictor of pain-related anxiety dimensions characterized by cognitive symptoms of anxiety. Overall, these findings reiterate the importance of anxiety sensitivity in understanding pain-related fear and anxiety, and suggest anxious and fearful responding can be predicted more accurately with higher levels of correspondence between a particular anxiety sensitivity domain and events that closely match that fear. PMID- 11400713 TI - Stability of emotions for traumatic memories in acute and chronic PTSD. AB - While memory for central factual information regarding an emotional event is considered to be relatively accurate, memory for emotions seem to be quite inaccurate (Christianson & Safer, 1995). We extended this line of research to examine memory for the emotional intensity surrounding a traumatic event (e.g., memory for the fear and horror of the event). We conducted a series of two studies. In Study 1, we examined memory for the emotional intensity of the traumatic event in recent sexual or non-sexual assault victims with acute PTSD at 2 and 12 weeks following the assault. In Study 2, we compared memory for emotional intensity in sexual and non-sexual assault victims with either acute or chronic PTSD at initial assessment and 12 weeks later. For both studies, participants were asked to recall general emotional intensity, fear intensity, and dissociative intensity of the traumatic event. Results suggested that memory for the fear of the traumatic event did not fluctuate over time. However, memory for the general emotional and dissociative intensity did fluctuate over time, decreasing for individuals with acute PTSD and increasing for individuals with chronic PTSD. PMID- 11400714 TI - Brief cognitive therapy for social phobia: a case series. AB - Social phobia is a common and disabling anxiety disorder. The most effective psychological treatments for social phobia are cognitive therapy and exposure. However, the degree of improvement across these treatments is variable, and their implementation is costly and time-consuming. This study aimed to conduct a preliminary clinical evaluation of the effectiveness of a brief, new form of cognitive therapy based on a recent cognitive model of social phobia. Six consecutively referred patients with social phobia were treated using established single case series methodology. Brief cognitive therapy was effective with all patients demonstrating clinically significant improvements in all measures. Treatment gains were maintained at follow-up. The mean number of treatment sessions delivered was 5.5 and improvements compare favourably with previous treatment studies. Brief cognitive therapy for social phobia appears promising and it is potentially cost-effective. Future randomised and controlled evaluations of this brief treatment are warranted. PMID- 11400715 TI - Worry propensity and the perception of risk. AB - This paper examines how the tendency to worry impacts risk estimation for an upcoming, stressful event. Students in a statistics class completed worry, anxiety, and subjective risk questionnaires on the first day of class. State anxiety and subjective risk questionnaires were again administered immediately before their first examination. Subjects showed significant increases in anxiety and the perceived risk of poor academic performance in the class. Risk domains unrelated to the examination (e.g., physical threats) remained stable across measurement periods. Regression analysis was used to determine the independent contribution of worry propensity in predicting elevations in the perceived risk of poor academic performance. The analysis showed that worry propensity predicted subjective risk bias, even after variance attributable to current mood and trait anxiety had been removed. Results from a second series of regression equations suggested subjective risk biases partially mediated the relationship between worry propensity and current mood. These results are used to argue that the elevations in state anxiety observed when worriers are confronted with a stressful event are largely a function of risk inflation. PMID- 11400716 TI - Validity of the parenting scale for parents of children with attention deficit/hyperactivity disorder. AB - The present study examined the validity of the parenting scale for parents of elementary school-aged children with attention-deficit/hyperactivity disorder (ADHD). Parents from 109 families with children who had been diagnosed with ADHD (106 mothers and 93 fathers) and from 70 families with non-problem children (69 mothers and 59 fathers) completed the parenting scale and reported on their children's behavior problems. Factor analyses revealed two interpretable factors for both mothers and fathers, corresponding to the overreactivity and laxness factors identified in previous studies of the parenting scale. Overreactivity and laxness scores were significantly higher for mothers and fathers of ADHD children than of non-ADHD children; this effect appeared to be accounted for by comorbid aggression and conduct problems among ADHD children. Results support the validity of the parenting scale for use with parents of ADHD children. PMID- 11400717 TI - Predictive validity of the overvalued ideas scale: outcome in obsessive compulsive and body dysmorphic disorders. AB - Overvalued ideas have been theoretically implicated in treatment failure for obsessive-compulsive disorder (OCD). Until recently, there have not been valid assessments for determining severity of overvalued ideas. One recent scale, the Overvalued Ideas Scale (OVIS; Neziroglu, McKay, Yaryura-Tobias, Stevens & Todaro, 1999, Behaviour Research and Therapy, 37, 881-902) has been found to validly measure overvalued ideas. However, its predictive utility has not been determined. Two studies were conducted to examine the extent to which the OVIS predicts treatment response. Study 1 examined the response to behavioral therapy in a group of participants diagnosed with OCD. Residual gain scores showed a significant correlation between treatment outcome for compulsions and pretreatment OVIS scores (28.1% variance accounted). Pretreatment OVIS scores were not significantly correlated with residual gains in obsessions (1.7% variance accounted). The predictive utility of the OVIS was superior to a single item assessment of overvalued ideas available on the Yale-Brown Obsessive Scale in predicting outcome for compulsions. For this item, the variance accounted for compulsions was 6.3% and for obsessions was 3.9%. Study 2 examined the response to behavioral therapy in a group of participants diagnosed with body dysmorphic disorder (BDD), a condition ostensibly linked to OCD and presumed to present with higher levels of overvalued ideas. Residual gains scores showed a significant relationship between obsessions and OVIS (accounting for 34.8% of the variance), but not for compulsions (10.2% variance accounted). As in Study 1, the predictive utility of the OVIS was superior to the single item assessment (with 0.2% variance accounted for compulsions, 2.4% variance accounted for obsessions). Taken together, the studies reported here show that this OVIS is predictive of treatment outcome, and the predictive value depends on which symptoms are used to assess outcome. Further, the scale is more effective in predicting outcome than a widely used single item assessment. PMID- 11400718 TI - Leukocyte rolling in rat mesentery venules: distribution of adhesion bonds and the effects of cytoactive agents. AB - A new method of analyzing in vivo measurements of leukocyte WBC rolling along venular endothelium (EC) has been developed to extract insightful information on the dynamics of WBC-EC bond formation and disruption. The rolling velocity of WBCs was obtained by intravital microscopy of rat mesenteric venules. For the "spontaneous" rolling observed following exteriorization of the mesentery, we estimated that the average distance between clusters of adhesion bonds which tether a rolling cell to the venular wall was about 2 microm, and that the average lifetime of a bond cluster at the trailing edge of the rolling cell, from its exposure to the tensile force to its release, was on the order of 0.05 s. Both the inter-cluster distance and the lifetime were significantly reduced by treatments with the chemoattractant N-formyl-methionyl-leucyl-phenylalanine and the cytokine interleukin-1, while the average lifetime of the stretched bond clusters was not significantly changed by treatment with the cytoskeleton modifying agents cytochalasin B and colchicine. Each of the four treatments significantly reduced the heterogeneity in the cell rolling velocity, presumably by the selective recruitment of WBC subsets from the circulating WBC population or by a reduction in the heterogeneity of endothelial adhesiveness. These results were analyzed in the context of in vitro data in the literature on molecular bonds of cell adhesion. The findings suggest that, in the case of "spontaneous" rolling, there are on average approximately 2-3 clusters of adhesion bonds between a rolling cell and the vessel wall, and approximately five bonds in each cluster. PMID- 11400719 TI - Modeling of early events in T cell signal transduction after controlled T cell activation by peptide major histocompatibility complex. AB - Calcium signaling was observed in murine T cells over time, starting at a precise moment of contact with a layer of fibroblasts expressing a stimulatory major histocompatibility class II-peptide complex. The contact was controlled by a film thinning apparatus. Intracellular calcium levels were followed with the ratiometric dye, Fura-2. The calcium response was highly synchronized and well fitted by a mathematical model. The model includes three components: a sequence of reactions occurring after T cell receptor (TCR) triggering; InsP3-mediated calcium release from intracellular stores (Meyer and Stryer, Proc. Natl. Acad. Sci. USA 85: 5051-5055, 1988); and slow changes in levels phospholipase C-gammal (PLCgammal) reflecting a decrease in receptor triggering rate. Each component in the model controls a different part of the response-the initial delay, the sharp rise, and the slow decay, respectively. Kinetic parameters determined from curve fitting were the initial delay in calcium signaling defined as the time when [PLCgammal] reached its half of its maximum (76 s), the coefficient characterizing calcium efflux from endoplasmic reticulum (ER) (2.86 microM s(-1), expressed per liter of cell volume), and a rate constant characterizing the diminishing yield of production of PLCgammal (0.00046 s(-1)) by active TCR. Only the parameter representing PLCgammal production varied much from cell to cell. PMID- 11400720 TI - Myofilament kinetics in isometric twitch dynamics. AB - To better understand the relationship between kinetic processes of contraction and the dynamic features of an isometric twitch, studies were conducted using a mathematical model that included: (1) kinetics of cross bridge (XB) cycling; (2) kinetics of thin filament regulatory processes; (3) serial and feedback interactions between these two kinetic processes; and (4) time course of calcium activation. Isometric twitch wave forms were predicted, morphometric features of the predicted twitch wave form were evaluated, and sensitivities of wave form morphometric features to model kinetic parameters were assessed. Initially, the impulse response of the XB cycle alone was analyzed with the findings that dynamic constants of the twitch transient were much faster than turnover number of steady-state XB cycling, and, although speed and duration of the twitch wave form were sensitive to XB cycle kinetic constants. parameters of wave shape were not. When thin filament regulatory unit (RU) kinetics were added to XB cycle kinetics, the system impulse response was slowed with only little effect on wave shape. When cooperative neighbor interactions between RU and XB were added, twitch wave shape (as well as amplitude, speed and duration) proved to be sensitive to variation in cooperativity. Importantly, persistence and shape of the falling phase could be strongly modified. When kinetic coefficients of XB attachment were made to depend on sarcomere length, changes in wave shape occurred that did not occur when only sliding filament mechanisms were operative. Indeed, the force-length relationship proved to be highly sensitive to length dependent XB attachment in combination with cooperative interactions. These model findings are the basis of hypotheses for the role of specific kinetic events of contraction in generating twitch wave form features. PMID- 11400721 TI - Improved in vitro quantification of the force exerted by the papillary muscle on the left ventricular wall: three-dimensional force vector measurement system. AB - Recent developments indicate that the forces acting on the papillary muscles can be a measure of the severity of mitral valve regurgitation. Pathological conditions, such as ischemic heart disease, cause changes in the geometry of the left ventricle and the mitral valve annulus, often resulting in displacement of the papillary muscles relative to the annulus. This can lead to increased tension in the chordae tendineae. This increased tension is transferred to the leaflets, and can disturb the coaptation pattern of the mitral valve. The force balance on the individual components governs the function of the mitral valve. The ability to measure changes in the force distribution from normal to pathological conditions may give insight into the mechanisms of mitral valve insufficiency. A unique in vitro model has been developed that allows quantification of the papillary muscle spatial position and quantification of the three-dimensional force vector applied to the left ventricular wall by the papillary muscles. This system allows for the quantification of the global force exerted on the posterior left ventricular wall from the papillary muscles during simulation of normal and diseased conditions. PMID- 11400722 TI - Mechanoelectric feedback in a model of the passively inflated left ventricle. AB - Mechanoelectric feedback has been described in isolated cells and intact ventricular myocardium, but the mechanical stimulus that governs mechanosensitive channel activity in intact tissue is unknown. To study the interaction of myocardial mechanics and electrophysiology in multiple dimensions, we used a finite element model of the rabbit ventricles to simulate electrical propagation through passively loaded myocardium. Electrical propagation was simulated using the collocation-Galerkin finite element method. A stretch-dependent current was added in parallel to the ionic currents in the Beeler-Reuter ventricular action potential model. We investigated different mechanical coupling parameters to simulate stretch-dependent conductance modulated by either fiber strain, cross fiber strain, or a combination of the two. In response to pressure loading, the conductance model governed by fiber strain alone reproduced the epicardial decrease in action potential amplitude as observed in experimental preparations of the passively loaded rabbit heart. The model governed by only cross-fiber strain reproduced the transmural gradient in action potential amplitude as observed in working canine heart experiments, but failed to predict a sufficient decrease in amplitude at the epicardium. Only the model governed by both fiber and cross-fiber strain reproduced the epicardial and transmural changes in action potential amplitude similar to experimental observations. In addition, dispersion of action potential duration nearly doubled with the same model. These results suggest that changes in action potential characteristics may be due not only to length changes along the long axis direction of the myofiber, but also due to deformation in the plane transverse to the fiber axis. The model provides a framework for investigating how cellular biophysics affect the function of the intact ventricles. PMID- 11400723 TI - Dielectrical model of cellular structures in radio frequency and microwave spectrum. Electrically interacting versus noninteracting cells. AB - A model of dielectrical properties of cellular structures of a tissue has been proposed. Cellular structures were presented as a composition of membrane covered spheres and cylinders that do not interact with each other. No restrictions were applied to the thickness of cellular membranes. The model was further generalized into a case of electrically interacting cells. The difference in dielectrical properties calculated with the model of electrically noninteracting versus interacting cells is inversely dependent on frequency. At biological values of cellular volume fraction near 0.7 (packed configuration) the difference is about 10%-15% in resistance and in epsilon' for frequencies near 0.1 MHz. Experimental data for myocardial tissue and theoretical data, for both interacting and noninteracting models, reasonably agree at frequencies of 1-100 MHz. PMID- 11400724 TI - Equivalent dipole source imaging of brain electric activity by means of parametric projection filter. AB - In the present study, spatial filters for inverse estimation of an equivalent dipole layer from the scalp-recorded potentials have been explored for their suitability in achieving high-resolution electroencephalogram (EEG) imaging. The performance of the parametric projection filter (PPF), which we propose to use for high-resolution EEG imaging, has been evaluated by computer simulations in the presence of a priori information on noise. An inhomogeneous three-concentric sphere head model was used in the present simulation study to represent the head volume conductor. An equivalent dipole layer was used to model brain electric sources and estimated from the scalp potentials. Various noise conditions were simulated and the parametric projection filter was compared with standard regularization procedures such as the truncated singular value decomposition (TSVD) and the Tikhonov regularization (TKNV). The present simulation results suggest that the proposed method performs better than that of commonly used inverse regularization techniques, such as the general inverse using the TSVD and the TKNV, when the correlation between the original source distribution and the noise distribution is low, and performs similarly when the correlation is high. A method for determining the optimum regularization parameter, which can be applied to parametric inverse techniques, has also been developed. PMID- 11400725 TI - Discrimination of anesthetic states using mid-latency auditory evoked potential and artificial neural networks. AB - This study was undertaken to determine whether artificial neural network (ANN) processing of mid-latency auditory evoked potentials (MLAEPs) can identify different anesthetic states during propofol anesthesia, and to determine those parameters that are most useful in the identification process. Twenty-one patients undergoing elective abdominal surgery were studied. To maintain general anesthesia, the patients received propofol (3-5 mgkg(-1) h(-1) intravenously). Epidural analgesia at the level of T4-5 blocked painful stimuli. MLAEP was recorded continuously with patients awake, during induction, during maintenance of general anesthesia, and during emergence until the patients were recovered from anesthesia. Latencies of the 5 MLAEP peaks and three peak to peak amplitudes were measured, along with hemodynamic parameters (heart rate, systolic, and diastolic arterial blood pressure). Four-layer ANNs were used to model the relationship between the parameters of the MLAEP and the four different states (awake, adequate anesthesia, during/before intraoperative movement, and emergence from anesthesia). The best identification accuracy was obtained using only the five latencies. The combination of five latencies and three amplitudes did not improve the identification accuracy. Use of the only the three hemodynamic parameters produced a much poorer identification. This study suggests that the MLAEP has useful information for identifying different anesthetic states, especially in its latencies. A nonlinear discrimination approach, such as the ANN, can effectively capture the relation between the MLAEP patterns and the different states of anesthesia. PMID- 11400726 TI - Separation of a phenol carboxylating organism from a two-member, strict anaerobic co-culture. AB - In a culture converting phenol to benzoic acid under anaerobic conditions and previously described as being constituted of only a Clostridium-like strain 6, another bacterium (strain 7) was observed. Each organism was enriched by centrifugation on a Percoll gradient. Strain 6 was purified by dilution and plating. Strain 7 did not grow on solid media, but a strain 7 culture, cleared of strain 6, was obtained by subculturing in the presence of ampicillin and by dilution. In fresh medium, phenol was transformed by the reconstituted co-culture but not by each strain alone. In a supernatant from a co-culture or from a strain 6 culture, strain 7 alone transformed phenol but not strain 6. Maintenance of an active strain 7 in fresh medium instead of co-culture supernatant became possible when phenol was replaced by 4-hydroxybenzoate (4-OHB), which is decarboxylated to phenol before being transformed to benzoate. Even with 4-OHB, the use of co culture (or strain 6 culture) supernatant resulted in faster transformation activity and growth rate. A phylogenetic analysis placed strain 7 in a cluster of uncultivated or nonisolated bacteria (92-96% homology). Strain 7 is also related to Desulfotomaculum, Desulfitobacterium, Desulfosporosinus, Moorella, and Sporotomaculum genera (87-92% homology). PMID- 11400727 TI - The effect of temperature on the fatty acids and isozymes of a psychrotrophic and two mesophilic species of Xenorhabdus, a bacterial symbiont of entomopathogenic nematodes. AB - In the first part of this study, generation times relative to temperature, together with cardinal and conceptual temperatures, were determined for four strains of Xenorhabdus bacteria that represented three geographically distinct species. The data showed that the NF strain of Xenorhabdus bovienii, like the Umea strain of the same species, is psychrotrophic, while Xenorhabdus sp. TX strain resembles Xenorhabdus nematophila All strain in being mesophilic. In the second part, the capacity of these bacteria to adapt to changes in temperature, shown by changes in fatty acid composition, was investigated. As temperature declined, the proportions of the two major unsaturated fatty acids, palmitoleic (16:1omega7) acid and oleic (18:1omega9) acid, increased significantly in all of the strains. The proportion of the prevalent saturated fatty acid, which was palmitic acid (16:0), decreased. In the All, NF, and Umea strains, myristic acid (14:0), margaric acid (17:0), cyclopropane (17:0c), and arachidic acid (20:0) decreased with decreasing temperature. In the third part of the study, the synthesis of isozymes in response to changing temperature was investigated. For the seven enzymes studied, the numbers for which isozyme synthesis was temperature related were as follows: five for Umea, four for All, three for NF, and two for TX. Where the study dealt with fatty acid composition and isozyme synthesis, the results show a broad capacity for physiological temperature adaptation among strains of different climatic origin. PMID- 11400728 TI - Utilization of herpes simplex PCR assays for cerebrospinal fluid in a pediatric health care setting. AB - An assessment was made of the utilization and impact of a diagnostic polymerase chain reaction (PCR) assay for the diagnosis of herpes simplex viruses (HSV) 1 and 2 in cerebrospinal fluid of children who attended a Canadian pediatric referral centre. One hundred and three assays were performed on specimens from 103 patients during the period August 1997 to September 1998. Patient ages ranged from newborn to 16 years. Indications for HSV PCR included seizures with or without fever (56.3%), aseptic meningitis (16.5%), and encephalopathy with or without fever (10.7%). Only 2 of 103 (1.9%) assays were positive, including one each for HSV1 and HSV2. Control specimens that were seeded with virus indicated inhibition for 24.3, 8.8, and 6.8% of assays for HSV1, HSV2, and both HSV1 and HSV2, respectively. The mean turn-around time for HSV PCR was 2.5 days, and 90.3% were completed in less than 5 days. Acyclovir was administered to 78.6% of the patients overall; the results of the HSV PCR impacted on the treatment courses for 36 individuals. Nevertheless, 16.5% of patients continued to receive extended courses of antiviral therapy despite negative HSV PCR assays. Although it is desirable to decrease the frequency of PCR inhibitions and to further decrease the interval to assay completion, HSV PCR does have a significant impact on antiviral use in this setting. PMID- 11400729 TI - Study of lipase-catalyzed hydrolysis of some monoterpene esters. AB - Studies of the hydrolysis of bornyl, citronellyl, geranyl, and terpenyl acetates by commercially available lipases of Candida rugosa, Rhizopus arrhizus, and Mucor miehei are presented. The hydrolysis of these monoterpene esters is investigated at various temperatures and pHs, and the time dependence of the percentage of esters hydrolysed is studied. The catalytic activities of the lipases in hydrolysing the esters are compared and, overall, it is observed that under the experimental conditions used the nonspecific lipase from C. rugosa produces the highest yields of the monoterpene alcohols in comparison to the primary-ester specific lipases such as R. arrhizus and M. miehei. A rate kinetic model has been used to understand the time dependence of the yield of the product acid. PMID- 11400730 TI - Cytokinin production by plant growth promoting rhizobacteria and selected mutants. AB - One of the proposed mechanisms by which rhizobacteria enhance plant growth is through the production of plant growth regulators. Five plant growth promoting rhizobacterial (PGPR) strains produced the cytokinin dihydrozeatin riboside (DHZR) in pure culture. Cytokinin production by Pseudomonas fluorescens G20-18, a rifampicin-resistant mutant (RIF), and two TnphoA-derived mutants (CNT1, CNT2), with reduced capacity to synthesize cytokinins, was further characterized in pure culture using immunoassay and thin layer chromatography. G20-18 produced higher amounts of three cytokinins, isopentenyl adenosine (IPA), trans-zeatin ribose (ZR), and DHZR than the three mutants during stationary phase. IPA was the major metabolite produced, but the proportion of ZR and DHZR accumulated by CNT1 and CNT2 increased with time. No differences were observed between strain G20-18 and the mutants in the amounts of indole acetic acid synthesized, nor were gibberellins detected in supernatants of any of the strains. Addition of 10(-5) M adenine increased cytokinin production in 96- and 168-h cultures of strain G20-18 by approximately 67%. G20-18 and the mutants CNT1 and CNT2 may be useful for determination of the role of cytokinin production in plant growth promotion by PGPR. PMID- 11400731 TI - Iron content of Streptococcus suis and evidence for a dpr homologue. AB - The type strain of Streptococcus suis was investigated for features that might help the organism to tolerate the H2O2 that is produced during growth. Enzyme assays, using soluble extracts, revealed that the type strain, which lacks catalase, lacks NADH peroxidase in both the mid-exponential and stationary phases of the growth cycle. Although iron could not be detected colourimetrically in dense cell suspensions, determination of the cellular iron content following growth to early stationary phase in the presence of 55FeCl3 demonstrated that S. suis does contain iron and hence is incapable of iron exclusion. Gene amplification, using oligonucleotide primers based on dpr of Streptococcus mutans, followed by nucleotide sequencing, revealed in S. suis, the presence of a gene that encodes a Dpr homologue. It is concluded that in S. suis, tolerance of H2O2 is due to iron sequestration by Dpr and the consequent effect of this process on the extent of Fenton chemistry. PMID- 11400732 TI - Production of transferrin receptors by Histophilus ovis: three of five strains require two signals. AB - Five strains of Histophilus ovis (9L, 642A, 714, 5688T, and 3384Y) were investigated with respect to iron acquisition. All strains used ovine, bovine, and goat transferrins (Tfs), but not porcine or human Tfs, as iron sources for growth. In solid phase binding assays, total membranes from only two (9L and 642A) of the five strains, grown under iron-restricted conditions, were able to bind Tfs (ovine, bovine, and goat, but not porcine or human). However, when the organisms were grown under iron-restricted conditions in the presence of bovine transferrin (Tf), total membranes from all strains exhibited Tf binding (as above); competition experiments demonstrated that all three Tfs (ovine, bovine, and goat) were bound by the same receptor(s). Membranes from organisms grown under iron-replete conditions in the presence or absence of bovine Tf failed to bind any of the test Tfs. An affinity-isolation procedure allowed the isolation of two putative Tf-binding polypeptides (78 and 66 kDa) from total membranes of strains 9L and 642A grown under iron-restricted conditions, and from membranes of all strains if the growth medium also contained Tf. It is concluded that all strains tested acquire Tf-bound iron by means of siderophore-independent mechanisms involving surface receptors analogous to the Tf-binding proteins (TbpA and TbpB) found in comparable organisms; although iron restriction alone is sufficient to promote the expression of these proteins by strains 9L and 642A, their production by strains 714, 5688T, and 3384Y appears to require two signals, iron restriction and the presence of Tf. PMID- 11400733 TI - Sulfite oxidation by iron-grown cells of Thiobacillus ferrooxidans at pH 3 possibly involves free radicals, iron, and cytochrome oxidase. AB - Thiobacillus ferrooxidans cells grown on ferrous iron oxidized sulfite to sulfate at pH 3, possibly by a free radical mechanism involving iron and cytochrome oxidase. A purely chemical system with low concentrations of Fe3+ simulated the T. ferrooxidans system. Metal chelators, ethylenediamine tetraacetic acid (EDTA), 4,5-dihydroxy-1-3-benzene disulfonic acid (Tiron), o-phenanthroline, and 2,2' dipyridyl, inhibited both sulfite oxidation systems, but the T. ferrooxidans system was inhibited only after the initial brief oxygen consumption. EDTA and Tiron, strong chelators of Fe3+, inhibited the oxidation at lower concentrations than o-phenanthroline and 2,2'-dipyridyl, strong chelators of Fe2+. Inhibition of Fe3+-catalyzed sulfite oxidation by EDTA and Tiron was instant, but the inhibition by o-phenanthroline and dipyridyl was briefly delayed, presumably for the reduction of Fe3+ to Fe2+. Mannitol, a free radical scavenger, inhibited both systems to the same extent. Cyanide and azide inhibited only the T. ferrooxidans system, suggesting a role of cytochrome oxidase. It is proposed that sulfite is oxidized by a free radical mechanism initiated by Fe3+ on the cell surface of T. ferrooxidans. Cytochrome oxidase is possibly involved in the regeneration of Fe3+ from Fe2+ by the normal Fe2+-oxidizing system of T. ferrooxidans. PMID- 11400734 TI - Monitoring of bacteria in acid mine environments by reverse sample genome probing. AB - A variety of microorganisms can exist in acid mine drainage (AMD) environments, although their contribution to AMD problems is unclear. Environmental strains of Thiobacillus ferrooxidans and Thiobacillus acidophilus were purified by repeated plating and single-colony isolation on iron salts and tetrathionate media, respectively. Thiobacillus thiooxidans was enriched on sulfur-containing media. For the isolation of Leptospirillum ferrooxidans, iron salts and pyrite media were inoculated with environmental samples. However, L. ferrooxidans was never recovered on solid media. Denatured chromosomal DNAs from type and (or) isolated strains of T. ferrooxidans, T. acidophilus, T. thiooxidans, and L. ferrooxidans were spotted on a master filter for their detection in a variety of samples by reverse sample genome probing (RSGP). Analysis of enrichments of environmental samples by RSGP indicated that ferrous sulfate medium enriched T. ferrooxidans strains, whereas all thiobacilli grew in sulfur medium, T. thiooxidans strains being dominant. Enrichment in glucose medium followed by transfer to tetrathionate medium resulted in the selection of T. acidophilus strains. DNA was also extracted directly (without enrichment) from cells recovered from AMD water or sediments, and was analyzed by RSGP to describe the communities present. Strains showing homology with T. ferrooxidans and T. acidophilus were found to be major community components. Strains showing homology with T. thiooxidans were a minor community component, whereas strains showing homology with L. ferrooxidans were not detected. PMID- 11400735 TI - Presence of double-stranded RNA and virus-like particles in Rhizopus isolates. AB - Fungal isolates belonging to four Rhizopus species were screened for the presence of double-stranded RNA (dsRNA) molecules. Five (two R. stolonifer, two R. microsporus, and one R. oryzae) of the 27 isolates examined harboured such genetic elements. Electrophoresis of the nucleic acids revealed five RNA patterns, with 1-5 discrete dsRNA bands. The molecular sizes corresponding to these bands were 2.2-14.8 kb. Gel electrophoresis of purified virus-like particles (VLPs) indicated only one capsid of similar size in all virus harbouring strains; when investigated by electron microscopy, they were found to be polyhedral VLPs 40 nm in diameter. In one of the R. microsporus isolates an uncapsidated large dsRNA segment (14.8 kb) was observed. No phenotypic differences were observed between uninfected and virus-harbouring Rhizopus isolates. PMID- 11400736 TI - Purification, cloning, and sequencing of an enzyme mediating the reductive dechlorination of tetrachloroethylene (PCE) from Clostridium bifermentans DPH-1. AB - An enzyme mediating the reductive dechlorination of tetrachloroethylene (PCE) from cell-free extracts of Clostridium bifermentans DPH-1 was purified, cloned, and sequenced. The enzyme catalyzed the reductive dechlorination of PCE to cis 1,2-dichloroethylene via trichloroethylene, at a Vmax and Km of 73 nmol/mg protein and 12 microM, respectively. Maximal activity was recorded at 35 degrees C and pH 7.5. Enzymatic activity was independent of metal ions but was oxygen sensitive. A mixture of propyl iodide and titanium citrate caused a light reversible inhibition of enzymatic activity suggesting the involvement of a corrinoid cofactor. The molecular mass of the native enzyme was estimated to be approximately 70 kDa. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-assisted laser desorption ionization-time of flight/mass spectrometry (MALDI-TOF/MS) revealed molecular masses of approximately 35 kDa and 35.7 kDa, respectively. A broad spectrum of chlorinated aliphatic compounds (PCE, trichloroethylene, cis-1,2-dichloroethylene, trans-1,2-dichloroethylene, 1,1 dichloroethylene, 1,2-dichloropropane, and 1,1,2-trichloroethane) was degraded. With degenerate primers designed from the N-terminal sequence (27 amino acid residues), a partial sequence (81 bp) of the encoding gene was amplified by polymerase chain reaction (PCR) and sequenced. Southern analysis of C. bifermentans genomic DNA using the PCR product as a probe revealed restriction fragment bands. A 5.0 kb ClaI fragment, harboring the relevant gene (designated pceC) was cloned (pDEHAL5) and the complete nucleotide sequence of pceC was determined. The gene showed homology mainly with microbial membrane proteins and no homology with any known dehalogenase, suggesting a distinct PCE dehalogenase. PMID- 11400737 TI - Rapid detection of Brevibacillus formosus BN53-1 in chicken feces. AB - Here we describe a rapid method for detecting the hydrogen sulfide-decomposing bacterium Brevibacillus formosus BN53-1 in chicken feces. The method, which can be adapted to the specific detection of a variety of useful eubacteria, is based on blot hybridization and polymerase chain reaction (PCR), and makes use of the genus or species hypervariable region of eubacterial 16S rDNA. The approximate limit of detection under the conditions we tested was 1.0 x 10(3) cells in 10 mg of chicken feces. PMID- 11400738 TI - Pine nuts: the mycobiota and potential mycotoxins. AB - The mycobiota of pine nuts was investigated. In total, 1832 fungi belonging to 31 species and 15 genera (Ascomycota, 2; Zygomycota, 3; mitosporic fungi, 10) could be isolated. Cladosporium spp. dominated the mycobiota with 685 isolations followed by Phoma macrostoma with 351 isolations. Overall, 16 potentially mycotoxigenic species were present on pine nuts. PMID- 11400739 TI - IgA and IgM anti-Naegleria fowleri antibodies in human serum and saliva. AB - Antibodies from IgA and IgM classes that recognize Naegleria fowleri (Nf) proteins were detected by the ELISA assay in serum and saliva from three groups of people: (i) subjects with upper respiratory tract infections (URTI) living in the parasite-endemic area, (ii) healthy persons from the same area, and (iii) healthy persons from a parasite-nonendemic area. In serum and in saliva the titers of IgA antibodies to Naegleria fowleri in the group of patients with URTI was significantly higher than that of the healthy group in the parasite-endemic area; also the titers of IgM antibodies in serum were significantly higher in patients. On the contrary, in saliva the antibodies were higher in healthy people from the parasite-endemic area. In all cases the subjects from the parasite nonendemic area had lower antibody titers in serum and saliva. PMID- 11400740 TI - Newborn screening as we move to the twenty-first century. PMID- 11400741 TI - Newborn screening for congenital adrenal hyperplasia in Sapporo City: sixteen years experience. AB - A screening program for congenital adrenal hyperplasia (CAH) in Sapporo began in 1982, 7 years prior to the introduction of the national program. Since its inception, testing has involved the detection of 17-hydroxyprogesterone (17-OHP) in dried blood samples, using ELISA. Up to the end of March 1998, of 298,731 newborn screened, second samples were requested in 1,723 cases (0.6%). This number included 789 newborns who weighed less than 2,000 gm at birth. A total of 14 cases were diagnosed with 21-hydroxylase deficiency (21-OHD). "Salt-wasting type (SW)" outnumbered "simple virilizing type (SV)" by 11:3. The ratio of male to female was a converse. but unrelated, 3:11. Our study from 1982-1997 revealed that the incidence of 21-OHD in Sapporo City was 1:21.338, markedly similar to the worldwide incidence of 1:15,000. In order to improve the program, other type of analysis are also currently in use and under evaluation. These include highly sensitive HPLC analysis for 17-OHP and molecular analysis to identify some mutations associated with the 21-OHD gene (CYP21). These methodologies are very useful for the confirmation of information acquired from dried blood specimens. PMID- 11400742 TI - Screening and diagnosis of congenital adrenal hyperplasia in Basilicata (Italy). AB - Research of the frequency of 21-OH enzyme deficiency, autosomal recessive disease, caused by aberrations in the short arm of chromosome 6 was performed in order to prevent CAH (Congenital Adrenal Hyperplasia) manifested by: 1) possible cerebral damage 2) errors of sex attribution 3) behavioral hyperandrogenism 4) metabolic damage. Radioimmunoassay was used where there is competition between a radioactive and a non-radioactive antigens for a fixed number of antibody binding sites. In an 18 month period of screening 6,000 newborns we found one positive case of CAH which we confirmed by dosaging steroids such as, 4-androstenedione, testosterone, ACTH, PRA and electrolytic activity on the serum. We ascertained that an incidence of 1:6,000 in a 18 month period is high enough to deserve attention. PMID- 11400743 TI - Newborn screening for lysosomal storage disorders. AB - Lysosomal storage disorders (LSD) represent a group of over 40 distinct genetic diseases with a total incidence of approximately 1:7,000 births. Bone marrow transplantation and enzyme replacement therapy are currently in use for the treatment of some disorders and new forms of enzyme and gene replacement therapy are actively being researched. The effectiveness of these therapies, particularly for the LSD involving the central nervous system and bone pathology, will rely heavily upon the early diagnosis and treatment of the disorder, before the onset of irreversible pathology. In the absence of a family history the only practical way to detect these disorders will be by a newborn screening program. One common feature of these disorders is an increase in the number and size of lysosomes within the cell from approximately 1% to as much as 50% of total cellular volume. Associated with this, is a corresponding increase in some lysosomal proteins. We propose that the measurement of one or more of these proteins in blood spots taken from Guthrie cards, will form the basis of a newborn screening program, for the detection of all LSD. We have identified a number of lysosomal proteins as potential markers for LSD. The level of these proteins has been determined in blood spots taken from Guthrie cards and in plasma samples from over 300 LSD affected individuals representing 25 disorders. Based on these results we have proposed a strategy for a newborn screening program involving a two tier system, utilizing time resolved fluorescence immunoquantification of the protein markers in the first tier, followed by tandem mass spectrometry for the determination of stored substrates in the second tier assays. PMID- 11400744 TI - Dilemmas in newborn screening. PMID- 11400745 TI - Pilot neonatal screening program for lysosomal storage disorders, using lamp-1. AB - We have demonstrated that the lysosome associated membrane protein (LAMP-1) is elevated in plasma from approximately 70% of lysosomal storage disorder patients. As part of the development of a newborn screening program for lysosomal storage disorders we have developed a first tier screening assay based upon the level of LAMP-I in blood spots taken from newborn Guthrie cards. To determine the effectiveness of the first-tier marker a prospective pilot Guthrie neonatal screening program for the identification of LSD was commenced in April 1998. Prior to commencement of the pilot program ethical approval was obtained and information leaflets regarding the neonatal screening of LSD were distributed to parents at the time of their infant's Guthrie collection. The LAMP-1 assay utilizes a chicken polyclonal and a mouse monoclonal in a sandwich time resolved fluorescent immunoassay. LAMP-1 blood-spot calibrators and quality control specimens were developed and shown to be stable and reproducible. To date 11,183 infants have been screened using LAMP-1. The population distribution is described with a median and 98th percentile of 220pg/l whole blood and 483microg/l whole blood respectively. Acceptable CV% for intra and inter assay of 8.9% and 10% respectively were obtained. PMID- 11400746 TI - Molecular screening for fragile X syndrome in Thailand. AB - Fragile X syndrome (FXS) is the most common form of inherited mental retardation. We screened for FXS in 237 Thai males (age < or = 15 years) with developmental delay of unknown cause. We found 16 (6.8%) to have FXS using standard molecular analysis. Wc then studied the extended families of these 16 FXS subjects and 4 other independently ascertained FXS cases. We found that there were at least 35 affected males and 8 affected females. In addition we found that there were at least 31 premutation carrier females and 4 premutation males. The CGG repeats numbers in these premutation individuals ranged from 60 to 125. By comparison, the normal CGG repeats were 19-50 with a heterozygosity of 67.2% in 337 randomly selected males. This study providcs insight into the high incidence of FXS in developmentally delayed Thai males and points the way toward the means of prevention of mental retardation by genetic counseling and prenatal diagnosis. PMID- 11400747 TI - Inherited metabolic disorders: from the newborn to the mother and beyond. PMID- 11400748 TI - Molecular diagnosis of inborn errors of metabolism. PMID- 11400749 TI - Inherited metabolic disorders in Thailand--Siriraj experience. AB - The incidence of inborn errors of metabolism (IEM) in Thailand is yet unknown. However, by estimation it is generally accepted to be 1 in 5,000. From a survey in 7 medical schools from different parts of the country and a large pediatric hospital in Bangkok, we found numerous cases of IEM nationwidc. Thesc were amino acid disorders, carbohydrate disorders, urea cycle defects, peroxisomal, lysosomal storage disorders, and many others. Since Thais are quite homogeneous in their genetic make-up; it is, therefore, very likely that IEM is much more prevalent than we realized. With the exception of thalassemias, IFAM is probably very common in Thailand and other countries in the Asia-Pacific region. IEM identified were amino acid disorders eg phenylketonuria, maple syrup urine disease: urea cycle disorders eg ornithine transcarbamylase deficincy (OTC), argininosuccinic lyase deficiency (ALD), argininosuccinic acid synthetasc deficiency (ASD); glycogen storage disorders eg Pompe's discase, Von Gierkc's; organic acid disorders eg, isovaleric acidemia, methylmalonic acidemia. Lysosmal storagc disorders identified were GM1 gangliosidosis, mucolipidosis II, Hurler, Hunter, Maroteaux-Lamy, Sialidosis (neuraminidase deficicncy), Sly, Scheie, Gaucher, Niemann-Pick, Sandhoff and many other neurodegeneraative disorders identified were rhizomelic chondrodysplasia punctata (RCDP) and Zellweger. Recently fatty acid oxidation disorders: MCAD, translocase deficiency and multiple carbosxylase deficiency 9biotinidase deficiency) were also identified. PMID- 11400750 TI - Plasma amino acid analyses in two cases of maple syrup urine disease. AB - Maple syrup urine disease is a rare inborn error of metabolism, characterized by elevated plasma levels of branched chain amino acids and urinary excretion of branched chain keto acids. Plasma amino acid levels in two subjects were followed by deproteinizing plasma, derivatizing the free amino acids with phenylisothiocyanate, and analysis by HPLC. The results indicate that valine, leucine and isoleucine are elevated in Maple syrup urine disease, and that leucine remains high even after dietary treatment. PMID- 11400751 TI - Ethical and social issues in newborn screening. PMID- 11400752 TI - Plasma amino acid and urine organic acid analyses of methylmalonic acidemia in a Thai infant. AB - Methylmalonic acidemia is an inborn error of organic acid metabolism resulting from defects in methylmalonyl CoA mutase. Analysis of plasma free amino acids in a 15-month-old Thai infant by HPLC showed marked elevation of glycine. HPLC analysis of urinary organic acids showed high levels of methylmalonic acid. PMID- 11400753 TI - Gaucher's disease;thirty-two years experience at Siriraj Hospital. AB - Gaucher's disease, a lysosomal disorder, is not a common disease in Thailand. During the period 1966-1998 we saw 20 patients with Gaucher's disease at the Department of Pediatrics. Siriraj Hospital. The patients came from different regions of the country but mostly from the central part of Thailand. There were 8 males and 12 females from 13 families of Thai, Thai-Chinese, Thai-Laos and Chinese-Chinese in origin. A history of consanguinity was present in 2 families. The age of onset was 2 months-4 years and the age when they were diagnosed was 4 months-15 years. The most common clinical features included splenomegaly, hepatomegaly, growth retardation, pallor, bleeding disorders and neurological abnormalities. The diagnosis was made by the clinical manifestations, hematologic complications and demonstration of Gaucher cells in the bone marrow and/or other tissues. In one family, the diagnosis was confirmed by evaluation of glucocerebrosidase activities in skin fibroblasts. The management of these patients was symptomatic ie packed red cell and platelet transfusion, splenectomy and other supportive measures. Most patients died of bleeding or infection at an early age. PMID- 11400754 TI - Screening for hyperhomocysteinemia in young adults with premature coronary artery disease. PMID- 11400755 TI - Studies of mass infant screening for Wilson disease by urinary ceruloplasmin. AB - We found a 4-year-old boy in the screened cohort, who showed a low ceruloplasmin (CP) in urine (17 ng/mg-creatinine) and in blood (0.6 mg/dl), but his urine copper was in the normal range. Furthermore this child was diagnosed as Wilson disease (WD) by genetic analysis. Although no significant correlation was observed between urine and blood levels of CP, it is conceivable that WD may be accompanied by very low concentration of urine CP. PMID- 11400756 TI - Persistent tyrosinemia detected by thin-layer chromatography. AB - Screening for tyrosinemia is not routinely performed worldwide. Using a low expense thin-layer chromatography (TLC) for amino acids we detected a high frequency of transient tyrosinemia with secondary hyperphenylalaninemia in some newborns. Serum follow up showed the need to introduce adequate therapy in these babies. PMID- 11400757 TI - Therapy of metabolic disorders with intravenous (IV) access ports and long term intravenous L-carnitine therapy. AB - With the expansion of newborn screening to include many organic acidurias and fatty acid oxidation defects, effective therapies of these disorders will be needed. Currently severe disorders such as methylmalonic and propionic aciduria. conventional therapy with diet and oral L-camitine often prove ineffective in preventing failure to thrive and recurrent metabolic decompensations. L-carnitine provides a natural pathway for removal of the toxic metabolites in these disorders and is life saving therapy but, with poor oral absorption (25%), it is difficult to supply adequate carnitine to meet the metabolic needs of these patients. Long term intravenous L-carnitine therapy, administered through a subcutaneous venous access port in 5 patients with organic acidurias [propionic aciduria (2), methylmalonic aciduria (2), 3 methylglutaconic aciduria(1)] resulted in improved growth, lower frequency of metabolic decompensations and increased tolerance of natural protein in the diet. An added benefit was the ability to initiate fluid. electrolytes, and antibiotics during metabolic decompensations at home thus averting hospitalizations. PMID- 11400758 TI - Detection of inherited metabolic disorders via tandem mass spectrometry in Thai infants. AB - From a retrospective study in Medical Genetics Unit, Department of Pediatrics, Siriraj Hospital Faculty of Medicine, Mahidol University in Bangkok (1983-1988), the estimated pediatric patients with clinically suspected IEM are approximately 2-4% of total annual pediatrics admission of 5,000 or more. This is, a low estimation since survey from all teaching hospitals in the country including the largest Children's Hospital in Bangkok indicated the presence of numerous IEM. However, most IEM were clinically diagnosed with limited laboratory facilities. We started a collaboration with Magee Womens Hospital of Pittsburgh and NeoGen Screening, USA; using tandem mass spectrometry to diagnose high risk infants and children for IEM from July 1993 to March 1998. Of total 146 samples sent, we detected numerous metabolic disorders (11.2%) eg phenylketonuria, organic acidemia, maple syrup urine disease, isovaleric acidemia, methylmalonic acidemia, albinism, translocase/carnitine palmitoyltransferase type II, G6PD deficiency and lysinuric protein intolerance. PMID- 11400759 TI - Detection of inborn errors of metabolism in Thai infants via gas chromatography and mass spectrometry. AB - We had studied inherited metabolic disorders at the Department of Pediatrics, Siriraj Hospital Faculty of Medicine, Mahidol University since 1987 using limited resources available and collaboration with other laboratories, both in Thailand and The United States. Since April 1998, we started a collaboration with MILS and Kanazawa Medical University, the Japan, studying inborn errors of metabolism in Asian Countries using urine filter paper and a new GC/MS method. We have since successfully discovered several patients with metabolic disorders. Out of 33 (high-risk) cases we sent for biochemical diagnosis (during April-July 1998), 13 abnormal results were found which is approximately 39.4%. Inherited metabolic disorders identified were as follows: medium-chain acyl CoA dehydrogenase deficiency (MCAD), multiple carboxylase deficiency (MCD), methylmalonic acidemia (MMA), Fanconi syndrome, galactosemia and neuroblastoma. PMID- 11400760 TI - Design of a pilot study to evaluate tandem mass spectrometry for neonatal screening. AB - In recent years, tandem mass spectrometry has generated great interest as a method for neonatal screening. The basic principle is electronically controlled separation of analytes by their mass-to-charge ratio. The advantage of this detection system is speed, the capability to analyze for many different compounds in a single analysis, and a minimal need for auxilliary assay reagents. The prevailing screening technique uses stable isotope dilution, butylesterification, and MS/MS analysis to quantify amino acids and acylcamitines in neonatal dried blood spot samples. This allows detection of more than 30 inborn errors of metabolism of amino acids, fatty acids, and other organic acids. In Denmark, a large-scale pilot study is being implemented to evaluate the screening potential of tandem mass spectrometry. National patient registers and medical records from clinical genetics units are used to identify cohorts of healthy infants and infants with selected inborn errors of metabolism. The neonatal screening samples of these infants are retrieved from a biobank and are assayed for amino acids and acylcarnitines using tandem mass spectrometry. This study yields decision values for neonatal screening, which will be evaluated in a subsequent 2-year prospective pilot study, offering the test to 140,000 neonates as a voluntary adjunct to the existing screening program. The organization consists of integrated units for neonatal screening and clinical genetics. The effect of the program will be assessed in terms of screening efficiency, cost and short term clinical outcome. PMID- 11400761 TI - Neonatal screening of phenylketonuria and congenital hypothyroidism in China. AB - Mass screening of newborns for some congenital metabolic diseases is a tremendous achievement in the field of preventive medicine. The pilot study of neonatal screening in China was started in 1981. According to the results of 1.1 million newborn screenings from 8 big cities in the past 6 years, the incidence of phenylketonuria was 1/14,767 and congenital hypothyroidism was 1/5,469. China is a vast country with great economic differences among the different regions, making neonatal screening difficult in some areas. The national neonatal screening coverage rate is only about 2 % at the moment, but in metropolitan centers like Shanghai, the coverage rate reached 95 % in 1997. PMID- 11400762 TI - Screening blood spots for argininosuccinase deficiency by electrospray tandem mass spectrometry. AB - Argininosuccinase deficiency is relatively more common in Saudi Arabia than other urea cycle detects (UCD) and its presentation is usually acute and virtually identical to the clinical presentation of other UCD. We developed a rapid, sensitive, and specific screening method for the diagnosis of argininosuccinase deficiency from blood spots. using electrospray tandem mass spectrometry. A 96 well microplate batch process is used for extraction of argininosuccinic acid (ASA), other amino acids and acylcarnitines (Rashed et al. 1995). ASA and other metabolites are derivatized to the corresponding butyl derivatives. The tris butyl ester of ASA (MH = 459.3) yields two major fragments at m/z 70 and m/z 144 under mild collision induced collision. montitored in the product ion spectrum using a narrow mass range (65-150 kDa). A processing algorithm "CAMPA" is used to automatically calculate the height ratios of selected masses and flags data files as "abnormal" when certain threshold is exceeded. The method is integrated with our existing 2-minute MS/MS method for profiling amino acids and acylcarnitines (Rashed et al. 1997). Using this approach for two years we diagnosed 16 ALD cases from 14 hyperammonemic infants, one high-risk newborn, and one from a regular newborn screening blood spot. PMID- 11400763 TI - Automated, simplified GC/MS data processing system for organic acidemia screening and its application. AB - Gas chromatography mass spectrometry (GC/MS) is widely used in diagnosis of organic acidemias. However, GC/MS has not yet become a routine laboratory test, because of the complexity in interpretation of GC/MS data. We developed a personal computer-based system of automated metabolic profiling and disease detection for the screening of organic acidemias by GC/MS. The data were processed after the GC/MS analysis of urinary organic acids. In this system, 130 kinds of metabolites and 25 disorders of organic acids were enrolled for the search and detection, respectively. Metabolites were identified with methylene unit values (MU). target ions (Q- and C-ions) and their intensity ratios, and semiquantified by peak relative area (%) of the Q-ions to that of an internal standard. Metabolites whose values exceeded the cutoff of the control table were flagged as abnormal. The diseases or pathological condition were automatically evaluated by combination of the abnormal compounds. In this system, index metabolites were categorized into three groups. "AND, "OR" and "NO". The groups, "AND" and "OR" comprised essential and optional compounds, respectively, for the specific diagnosis. The third group, "NO", included compounds which must be absent to reach a diagnosis. We compiled data of MU values and mass spectrum of 130 kinds of index metabolites, and tested the usefulness of this system by analysis of 74 patients with 19 kinds of diseases. In all cases, at least a correct diagnosis could be found among the disease names outputted. We have successfully applied this to a pilot neonatal screening by GC/MS in our regional area, and acylglycine analysis by the stable isotope dilution method with tert butyldimethylsilyl derivatization. With our system, many people can attend for screening programs using GC/MS. PMID- 11400764 TI - Screening pregnancies at risk for single gene disorders. PMID- 11400765 TI - Genetic counseling after unexpected cytogenetic findings on prenatal diagnosis. PMID- 11400766 TI - Prenatal prediction of spinal muscular atrophy by SMN deletion analysis. AB - The objective of this study was to provide prenatal prediction of spinal muscular atrophy (SMA) by survival motor neuron (SMN) gene deletion analysis and genetic counseling in families with previous child affected with SMA. The SMN gene is absent or interrupted in approximately 95% of SMA patients independence of clinical severity. We study four families with one previous child affected in each by performing the SMN deletion analysis in the index case. When a homozygous deletion in exon 7 or exon 8 is found, we offer prenatal prediction to the family. All four index cases had homozygous deletions of the SMN gene. Prenatal diagnosis by amniocentesis was performed in all pregnancies. Two pregnancies were positive for the homozygous deletion of the SMN gene, non-directive counseling was given and the two pregnancies were terminated. The other two pregnancies showed no deletion of the SMN gene. The unborn child is yet to be followed up. The prenatal prediction of SMA shows considerable requirements and potential effectiveness in prevention of the SMA in families at risk which cut the cost of care in this incurable disease. PMID- 11400767 TI - HIV screening in pregnant women. AB - Abstract. The following recommendations are made for prenatal screening for HIV infection 1) Routine voluntary screening for HIV infection in all pregnant women is feasible and worthwhile. 2) Every seropositive result should be repeated for confirmation before coming to a definititve conclusion to avoid a misdiagnosis. 3) Fetal blood sampling in the mid-trimester for PCR and p24 antigen assay seems inappropriate as a diagnostic tool for in utero HIV infection of the HIV seropositive pregnant women as the infection mostly occurs after this time. 4) Routine screening of seronegative pregnant women should be repeated during the third trimester to detect seroconversion since this offers a chance for antiretroviral administration to the seroconverted pregnant women for reduction of perinatal transmission. 5) There should be available the appropriate back up services for seropositive pregnant women. There is sufficient evidence indicating a higher vertical HIV-1 transmission rate in the last trimester and during labour compared with the first and second trimesters. Antiretroviral therapy either single or in combination given to the mother during the last trimester and delivery can reduce the viral load in the maternal circulation. Vertical HIV-1 transmission during delivery can be minimized by appropriate timing and route of delivery. Elective Cesarean section before the onset of labour with an intact bag of forewaters provides the least mother-to-fetus microtransfusion compared to other modes of delivery. Since an effective combination of HIV-1 immunoglobulin and HIV-1 vaccine given to the HIV-1 exposed newborns to prevent HIV-1 transmission similar to the viral hepatitis B model is not firmly established at present, postexposure antiretroviral prophylaxis and nonbreast-feeding are advocated for infants born from the HIV-1 infected mothers. In cases of advanced stage of maternal HIV-1 infection, and in developing areas where malnutrition prevails, an adequate supply of essential micronutrients is proposed as an adjunctive measure to reduce HIV-1 perinatal transmission. PMID- 11400768 TI - HIV antibody measurement of dried blood sample and its application to sero prevalence surveillance in pregnant woman. PMID- 11400769 TI - Prenatal ultrasound diagnosis in Thailand. AB - Ultrasound technology has been extensively employed in obstetric and gynecologic practice for several decades. It has been used not only in gestational age estimation, placental location, amniotic fluid assessment but also in antenatal diagnosis of fetal abnormalities of various systems. The incidence of Congenital Heart Disease (CHD) is currently estimated to be 8 to 9 out of every 1,000 live births. Congenital heart disease is thought to be a multifactorial disorder in over 90% of the cases. Pioneer studies on the ultrasound investigation of the heart were reported in the early 1970s. Since the introduction of high resolution real-time ultrasound in the late 1970s, reports on ultrasound assessment of fetal cardiac anatomy and function have been appearing with increasing frequency in both the obstetric and cardiologic literature. At present, fetal echocardiography is a well established technique for the prenatal diagnosis of CHD. However, the distribution of the technique is still limited as it requires both a very experienced operator and meticulous scanning. Screening the entire obstetric population does not appear possible at present. Therefore fetal echocardiography is as necessarily directed toward selected pregnancies carrying a higher-than normal risk of fetal cardiac anomalies. The progressive developments in cardiac ultrasound during the past two decades have substantially altered the practice of perinatologists, obstetricians and cardiologists. Total cardiac ultrasound studies are now becoming the primary armamentarium in the diagnosis of fetal congenital heart diseases. Since a 92% sensitivity of 4-chamber view in screening fetal heart diseases was first reported in 1987, 4-chamber view has been widely recommended for routine use in fetal sonographic examinations. Recent studies have demonstrated somewhat variable results and have suggested incorporation of the out flow should thoroughly understand the advantages and limitations of fetal cardiac scanning especially the 4-chamber view in order that the appropriate information and management plan can be offered to the patients. Fetal cardiac scanning is well recognized as one of the most tedious scanning of all fetal organ systems. With a tremendous advancement in ultrasound resolution including color imaging capacity several kinds of fetal cardiac anomalies could be diagnosed antenatally which inevitably allows more proper management plan for couples engaged in these. PMID- 11400770 TI - Polymerization of fetal fibrin and alterations in fibrin network characteristics during diabetic pregnancy. AB - The present study compares plasma fibrin network characteristics of fetal blood and that of normal and diabetic pregnant women. Plasma fibrinogen concentration, clotting curves, mass-length ratio of the fibrin fibers, gross permeability and tensile strength of the networks, have been measured. Plasma glucose and glycated hemoglobin are used as glycemic index in diabetic gestational women. The fetal plasma has a lower concentration of fibrin and exhibits delayed clotting, the networks are made up of thinner fibers, are more cross-linked, have lower permeability, and increased tensile strength than in normal adults. The tensile strength of the networks prepared from the plasma of diabetic gestational women are more highly crosslinked than those made from plasma of normal women of corresponding length of pregnancy. The fibrin fiber-thickness is increased during the first and second trimester but is significantly reduced during the third trimester in the diabetic gestational women. The gross permeability of the networks is significantly reduced during the second and third trimester in the diabetic gestational women. The SDS-PAGE shows characteristic pattern of alpha, beta, and gamma-polypeptides in both normal and diabetic gestational women. PMID- 11400771 TI - Newborn screening in Singapore. AB - Neonatal screening in Singapore for G6PD deficiency started in 1965. Screening for congenital hypothyroidism started in 1981 as a pilot research program and by 1990, it became nationwide. Screening for congenital hypothyroidism is by the measurement of TSH in the cord serum with recall of those exceeding the 99th percentile, by about 3-4 days of life. Treatment is usually started within a week of life. Over 400,000 newborns have been screened and the screening rate has been about 99.95%. The incidence rate is about 1 in 3,000. G6PD activity in the cord blood is measured using semi-quantitative rapid screening tests. Those identified are physically protected from environmental triggers by keeping them in hospital for a variable period of time. Parents are counseled. Data obtained from 22,830 newborns from the National University Hospital revealed incidence rates of 1.62% in all newborns, 3.15% in males and 0.11% in females. The Chinese and Malay males had a higher (3.94% and 2.95%) incidence respectively when compared to the Indian males with (0.66% incidence). The application of the preventive measures has resulted in no report of kernicterus in the last 20 years. Our efforts are now focused on minimizing the recall rate in the case of hypothyroidism screening, reducing the period of stay in hospital in those with G6PD deficiency and considering the introduction of a PKU screening program in Singapore. PMID- 11400772 TI - Neonatal screening for hypothyroidism at a university hospital in Thailand. AB - Neonatal screening is an essential program for early identification of congenital hypothyroidism. Between July 1991 and May 1998, 37, 262 infants born at Chulalongkorn Hospital were enrolled to the screening program. Blood TSH levels were determined on infants at > or = 48 hours after birth. They were performed in dried blood sample taken by heel prick on filter papers. TSH levels were measured by fluoroimmunoassay from July 1991 to December 1997 and by immunoradiometric assay from January to May 1998. Infants with TSH screening level higher than the cut off level (20 mu/l) were recalled for re-evaluation which consist of complete physical examination and blood test for serum T4 and TSH. Bone age determination and thyroid scan using technetium-pertechnetate were performed if the serum T4 and TSH levels were abnormal. The recall rate was 0.28% and response rate was only 69%. Primary congenital hypothyroidism was diagnosed in 15 infants. Prevalence was 1: 2,484. Among these infants, 8 had ectopic thyroid, 3 had normal glands and 3 were athyrotic. One infant died before the thyroid scan could be performed and did not receive treatment. The median age at initiation of thyroxin therapy was 29 days (range, 20-67 days). The follow up result was satisfactory. This study demonstrated the potential vulnerability of congenital hypothyroid screening program in Thailand. Improvement of parents' education, communication and monitoring should be emphasized for a large screening program. PMID- 11400773 TI - Neonatal screening program in Rajavithi Hospital, Thailand. AB - A pilot study was conducted in order to identify the cases and determine the incidence of congenital hypothyroidism, phenylketonuria (PKU) and glucose-6 phosphate dehydrogenase (G6PD) deficiency in the newborn infants born at Rajavithi Hospital. During May 1995 - July 1998, 32,407 out of 49,092 (66%) infants were screened for congenital hypothyroidism by measuring thyroid stimulating hormone (TSH) by radioimmunometric assay method. Seven cases of congenital hypothyroidism were identified (incidence of 1 : 4,629 live births). The recall rate was 0.66%. The screening for PKU was done on 17,421 out of 29,443 (59.14%) infants by measuring phenylalanine level by Guthrie method during June 1996 to July 1998. There was no PKU found. From January 1996 to July 1998. 24,714 newborn infants were screened for G6PD deficiency by fluorescent screening technique. The total incidence of 5.13% was found. The incidence in males and females was 9.13% and 1.66% respectively. This study confirms the benefit of the screening program in early detection and treatment of the disorders. PMID- 11400774 TI - Neonatal screening for congenital hypothyroidism and phenylketonuria at Siriraj Hospital, Mahidol University, Bangkok, Thailand--a pilot study. AB - A newborn screening program for congenital hypothyroidism (CH) and phenylketonuria (PKU), a pilot study, was initiated at the Medical Genetics Unit, Department of Pediatrics, Siriraj Hospital Faculty of Medicine, Mahidol University in Bangkok, Thailand from January 1994 to December 1998, using dried blood spots (DBS). A total of 18,739 infants (out of 85,150 livebirths) were screened (22 % coverage). Three cases of congenital hypothyroidism (CH) were identified (incidence of 1: 6,246, livebirths), by enzyme linked immunosorbent (ELISA) and fluoroimmunoassays using a cut-off level of TSH >20 microlU/ml: the recall rate of 0.24%. The screening for PKU was done by fluorometric (Guthrie) and enzyme linked immunosorbent (ELISA) methods; using cut-off levels of phenylalanine > 4 mg/dl and > 3.6 mg/dl respectively, with a recall rate of 0.13%. There was no PKU found. Our study, a voluntary program, emphasizes the importance of parental education and consent; specimen collection and handling; appropriate follow-up and referral to specialists for treatment and counseling. Routine newborn screening for CH and PKU is being established to ascertain the maximum coverage, using recommendations and guidelines from this pilot study. PMID- 11400775 TI - A nationwide follow-up system is essential for a neonatal screening program. AB - A long-term follow-up study gives many benefits to patients detected by neonatal screening, their families, and all members who are involved in neonatal screening program. However, the follow-up system is not functioning satisfactorily enough in Japan. A nationwide long-term follow-up system needs to be established immediately to make the neonatal screening system complete in Japan. PMID- 11400776 TI - Neonatal screening in Australia. PMID- 11400777 TI - International cooperation in neonatal screening: technical training course for newborn and infant screening. AB - We report the outline and results of our experience with a group training course of neonatal screening for health care professionals in developing countries. Sapporo City Institute of Public Health (SCIPH) has been offered a training course on neonatal screening once a year since 1991 under the Technical Training Program of the Japan International Cooperation Agency (JICA). The aims of this training course are to enhance the participants' technical knowledge and skills, and also to deepen their understanding of the principle of neonatal screening as well as the relevant diseases. Lectures and laboratory practice on phenylketonuria (PKU), congenital hypothyroidism (CH), congenital adrenal hyperplasia (CAH) and neuroblastoma are included in the 3-month program. After the completion of the training, participants are expected to play a major role in establishing and expanding neonatal screening system in each of their countries. We have received a total of 67 participants from 25 countries until March 1998: 58 pediatricians; 2 gynecologists; 6 biochemists; 1 administrative officer. After they returned to their countries, 11 engaged in neonatal screening and started PKU and CH screening in their institute, city or province in Argentina, Brazil, Mexico, Peru and Thailand. We believe that these results fulfilled our objectives. Also, for follow-up, SCIPH has been giving information and consultation to the participants on requests. This international cooperation network could also benefit our present network of the International Society Screening in the future. PMID- 11400778 TI - Newborn screening (NBS). Everyone screens for something! Pitfalls for the unwary! PMID- 11400779 TI - A comprehensive screening program in South Brazil. AB - We present the experience and figures about a screening program in South Brazil carried on in Porto Alegre, capital of the Southern Brazilian State. We present the tests performed routinely in our laboratory, the prevalence of some diseases and tests for infectious diseases to be added in the most comprehensive regional program in our country. PMID- 11400780 TI - Screening of the most common medium-chain acyl CoA dehydrogenase (MCAD) deficiency mutation (K329E) in the Czech newborn population. AB - Medium chain acyl-CoA dehydrogenase (MCAD) deficiency is the commonest inherited disorder of fatty acid oxidation. Most clinically ascertained cases are caused by thc point mutation K329E in MCAD gene. The frequency of this mutation as determined by usc of dried blood spots on Guthrie cards and the PCR NeoI digestion method. Using molecular ncwborn screening we found no K329E homozygote and 14 K329E heterozygotes in 2,826 newborns from Moravian area of the Czech Republic. Lower frequency of K329E carriers (1/202)) suggests that the incidence of MCAD deficiency will be probably lower in our population than we expected. PMID- 11400781 TI - Establishment of screening systems in East Asia. PMID- 11400782 TI - Neonatal screen of the world in the 21st century: the Mediterranean area experience. PMID- 11400783 TI - Newborn screening for PKU and congenital hypothyroidism in Latvia. AB - The newborn screening program in Latvia was started in 1980 as pilot study project. Mass screening for phenylketonuria (PKU) in the whole republic was started in 1987, but for congenital hypothyroidism (CH) it begun in 1996. Last two years the Latvian State Medical Genetics Center (SMGC) screened nearly 25,000 newborn dried blood specimens per year. Approximately 25% are repeat and control specimens. Efficiency of the screening program is nearly 97%. Fluorometric method is used for PKU screening, followed by amino acid analysis (HPLC) and analysis of DNA samples for mutations of the PAH gene. Most of the 51 Latvian PKU patients detected from 1980 to 1998 were selected by neonatal screening (38), while the others (13) were diagnosed during genetic counseling. The incidence of PKU in Latvia is 1:8,700 births. CH screening is based on measurement of thyroid stimulating hormone (TSH), using fluoroimmunoassay method, with cut-off value 10 mlU/l, as the primary screening test, and T3 and T4, as confirmatory tests for diagnosis. From 38,684 newborns 37,380 were screened for CH in 1996 and 1997. From these 1,438 (3.8%) newborns were recalled because of initial elevation of TSH level. From these 85 (0.2%) newborns had elevated TSH level during first two weeks of life and were referred to SMGC for repeat testing and follow-up. The diagnosis of CH has been confirmed in 6 babies. The incidence of CH in Latvia is about 1:6,450. PMID- 11400784 TI - Screening for congenital toxoplasmosis in Brazil. AB - Toxoplasmosis acquired at the end of the pregnancy can produce ocular and neurological sequelae. The detection of IgM anti-Toxoplasma gondii in dried blood spots of newborns is an important tool for early treatment and prevention. In this study we showed results of a pilot study about the prevalence of congenital toxoplasmosis in Brazil and concluded that this disease should be included in the menu of neonatal screening programs. PMID- 11400785 TI - The management of phenylketonuria (PKU). PMID- 11400786 TI - A clinical investigation of 228 patients with phenylketonuria in mainland China. AB - Neonatal screening for phenylketonuria (PKU) in Mainland China was started late and is incomplete. At the present time, it only covers 1-2% of the whole nation. There are about 1,500-2,000 PKU patients born in China annually. In this study, 228 PKU patients who were followed in our hospital were reviewed. Only 19 (8.3%) patients were detected by neonatal screening. Two hundred and nine (91.7%) cases were defined by pediatricians at the age of 1 month to 22 years, among them 94.5% had mental retardation, 48.9% developed epilepsy, and 22.9% gave up the treatment because of financial difficulty and loss of confidence in recovery. To help all of the patients, the only feasible way is the popularization of nationwide screening. PMID- 11400787 TI - Life-long treatment for phenylketonuria. AB - Life-long low-phenylalanine diet is recommended as an additional determinant to achieve the maximal success in the treatment of phenylketonuria (PKU). In this context education of teenage patients with PKU is essential. PMID- 11400788 TI - Molecular studies and prenatal diagnosis of phenylketonuria in Chinese patients. AB - Phenylketonuria (PKU) is one of the most common inborn errors of metabolic disorders. Although PKU induced mental retardation can be prevented after neonatal screening by following treatment with low phenylalanine diet, some parents are seeking prenatal diagnosis. We screened for mutations in exon 3 and 7 of the PAH gene using the DGGE and restriction enzyme method, in combination with STR linkage analysis. Prenatal diagnosis was carried out in 8 PKU families. With this strategy, we are able to make prenatal diagnosis in about 65-70% PKU families. All diagnosis was confirmed in the newborn. PMID- 11400789 TI - Mass screening of galactosemia: improved Beutler Test using automated quantitative fluorescence assay. PMID- 11400790 TI - Mass newborn screening for glucose-6-phosphate dehydrogenase deficiency in Singapore. AB - Recognition of kernicterus as a significant contributor to newborn mortality and morbidity in G6PD deficient babies and the ease with which G6PD deficiency could be detected and kernicterus prevented by avoidance of triggers, led to the establishment of mass newborn screening for G6PD deficiency in 1965. G6PD deficient newborns are identified within a day of birth by measuring the enzyme activity in cord blood. They are then physically protected from triggers by keeping them in the hospital for the first 2 weeks of life after their parents are counseled. Enzyme activity is measured using Wong's in-house modification of the Bernstein's technique or the BM G6PD Deficiency Screening test based on the Beutler assay. Close to 1.6 million newborns, representing practically 100% of all births have been screened. Analysis of data from 22,830 newborns at the National University Hospital reveals an incidence of 1.62% in all newborns. 3.15% in males and and 0.11% in females. A distinct racial variation in the incidence of deficiency was observed in males: Chinese 3.94%, Malays 2.95% and Indians 0.66%. Intermediate deficiency was most frequently identified (1.83%) in Chinese females. With the preventive measures. the incidence of kernicterus has dropped dramatically and there has been, over the last 20 years, no reported cases of kernicterus in newborns with G6PD deficiency. We are now looking at issues like comparing different assay techniques and determining a shorter period of stay in hospital. We believe that all Asians, especially Chinese babies, should be screened for G6PD deficiency, irrespective of which country the child is born. PMID- 11400791 TI - Neonatal screening for glucose-6-phosphate dehydrogenase deficiency in Taiwan. AB - Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzymopathic disease in Taiwan. The mass neonatal screening of G6PD deficiency by fluorometric spot test in Taiwan was started with a pilot program in 1984. The nationwide screening was started on July 1, 1987, and a follow-up system comprising of eighteen referral hospitals, including outlying islands, was organized for confirmatory test, medical care and genetic counseling. From July 1987 to December 1997, 2,971,192 heel blood samples collected on filter paper from 1,143 delivery units were screened by four neonatal screening centers. 46,570 cases were confirmed as G6PD deficiency is estimated to be around 2.1% (male 3.1%, female 0.9%) in Taiwan. The coverage rate of neonatal screening was 99% in 1997. To assess the reliability of the confirmatory test, an external quality assurance (QA) program for G6PD assay was developed. Periodically, 3 or 5 lyophilized quality control materials with different activities of G6PD were sent to each referral hospital by speed post delivery in dry ice. From January 1988 to June 1998, 85 QA services were performed. Two hundred and seven (13.5%) abnormal QA results were found, which were attributed to clerk (11.6%), procedural (16.4%), and instrumental errors (47.3%). In aid to confirm G6PD deficiency, a method to detect the G6PD mutation by using the dried blood samples was developed. The frequencies of the mutant alleles in Taiwan were determined to be 46.8% (1376G > T), 16.2% (1388G > A), 7.9% (95A > G), 6.5% (493A > G), 5.6% (392G >T), 4.6% (1024C > T), 0.5% (487G > A) and 0.5% (519C > G), respectively. PMID- 11400792 TI - Glucose-6-phosphate dehydrogenase deficiency in Thailand; its significance in the newborn. AB - The prevalence of G6PD deficiency in Thai males ranges from 3-18% depending upon the geographic region. G6PD "Mahidol" (163 Gly --> Ser) is the most common variant found in the Thai population. Almost all affected Thai individuals are not anemic and are asymptomatic. Severe acute intravascular hemolysis is occasionally seen, for instance, in those cases who have a viral infection, bacterial infection or have been exposed to chemicals or drugs. In Thailand, diagnosis of G6PD deficiency is usually made only in symptomatic cases. Neonatal screening of G6PD deficiency is not practiced nationwide, though studies have been done in several institutes. The assessment of G6PD activity in the newborn is mostly in order to find out the cause of neonatal jaundice. In our experience and that of others. G6PD deficient newborns are more prone to develop neonatal jaundice which is, on its own, no more severe than jaundice from other causes. Kernicterus due to G6PD deficiency, though still seen, is now very rare. Awareness of the hazard of hyperbilirubinemia, whatever the cause, along with active management is needed to prevent the occurrence of kernicterus. Neonatal screening is useful to detect abnormalities in the newborn. Weighing of the cost and benefit of neonatal screening should be made and the families of patients should be offered proper education and counseling to help them understand their babies' condition. PMID- 11400793 TI - Adequacy and pitfalls of G6PD deficiency counseling in Hong Kong. AB - Glucose-6-Phosphate-Dehydrogenase (G6PD) deficiency is common in Hong Kong with an incidence of 4.5% in male and 0.36% in female (Lo et al. 1996).The Neonatal Screening Unit of Clinical Genetic Service started its territory-wide neonatal screening program for G6PD deficiency and congenital hypothyroidism in 1984 (Lam et al, 1986). Because of insufficient manpower and resource, we have been giving health counseling on the phone to parents of G6PD deficient babies and then refer them to nearby maternal and child health centres for monitoring of jaundice. The disease, mode of inheritance, recurrence risk and the precaution against certain medicines (Chan 1996) and chemicals are explained. The purpose (Lam, 1994) is to reassure the parents that their G6PD deficient babies can be as normal as everyone and that they can have normal life. Nevertheless, it has not been established whether telephone counseling has any effect on the affected family in the form of psychological trauma (Marteau, 1989; Fyro, 1987; Li et al, 1996) or significant influence on the decision on future reproduction. This study tried to evaluate the service from the parents' point of view by 1) gathering information on parents' awareness and perception of G6PD deficiency, 2) determination of parents' attitude towards the telephone counseling, and 3) finding out the effect of G6PD deficiency on parents' decision on future reproduction. Over 300 parents were contacted by telephone, and were asked to respond to questions on a questionnaire . The telephone interview focused on parents' understanding of G6PD deficiency, their attitude towards this disease and the possible effect on future reproduction decision. Results showed that over 90% of cases that we had counseled attended the maternal and child health centres. Most of them accepted the presence of G6PD deficiency in their family which did not affect their decision on future pregnancy. Telephone counseling failed to establish a helping relationship with the parent as face to face counseling was more personal. The findings revealed that though telephone counseling had its shortcoming it served the target group effectively. Telephone counseling is still the method of choice for the G6PD deficiency counseling in this locality. PMID- 11400794 TI - A simple, rapid fluorometric assay for the determination of glucose 6-phosphate dehydrogenase activity in dried blood spot specimens. AB - Glucose 6-phosphate dehydrogenase (G6PD) deficiency is the most common enzymopathy of human beings and is the most common cause of jaundice and acute hemolytic anemia in South East Asia. The deficiency causes acute hemolytic anemia following ingestion of 6-amino quinoline antimalarials, phenacetin, and other substances. The rapid identification of infants or patients with this deficiency would help to prevent their exposure to these substances and subsequent risk to health. The assay is relatively simple. A 3mm punch from a dried blood spot sample is placed in a well of a black fluorescent microtiter plate containing calibrators and controls in duplicate. 100 microl of reagent is added and the sample is allowed to react for 30 minutes at ambient temperature after which 200 microl of stop reagent is added. The plate may be read immediately or up to one hour in a fluorescent reader (ex 355 nm: em 460 nm). Glutathione. ascorbate and bilirubin do not affect the assay. hemoglobin does quench the fluorescence by about 1.1 fluorescence units/g/dHb. This would not cause any false negatives and deficients would not be missed. G6PD activity in whole blood normal samples was examined at -20, 6 and 37 degrees C over 14 days. The samples lost about 20% activity after 48 hours and 31% by the end of 14 days. The samples stored at -20 degrees C and 6 degrees C remained relatively stable over this period. In a preliminary study eight diagnosed G6PD deficient samples had a mean value of 2.0 U/gHb (range 0.8 to 4.4) and fell within 3 SD units of the mean. Forty one normal samples had a mean of 6.6 micromol/min/gHb. Only one sample with a low hemoglobin level fell outside of 3 SD units of the mean. The Wallac assay was compared to the Sigma G6PD assay and although the values appeared lower at normal levels, the deficient samples compared well. PMID- 11400795 TI - G6PD deficiency in an unselected Brazilian population. AB - Data about glucose-6-phosphate dehydrogenase (G6PD) deficiency are not available in Brazil, a country characterized by a great mix of races. The disease is associated with ethnic groups. High prevalence (5 to 25%) has been reported in Africa, Asia, Middle East and the Mediterranean. We present here the first report of our one year experience testing for G6PD in an unselected population in the south of Brazil. PMID- 11400796 TI - Detection and structural analysis of abnormal hemoglobins found in Thailand. AB - In Thailand, some 20 different abnormal hemoglobins have been found in the last 30 years. Most are rare except for Hb E and Hb Constant Spring, found with frequencies of 10-53% and 1-8% respectively in different parts of the country. Most mutations are point mutations, but C-terminal elongations and crossing-over are also found. Most mutations do not cause clinical problems, but some can give rise to mild thalassemia syndromes, or cause problems in association with thalassemia. Abnormal hemoglobins may often be diagnosed by electrophoresis, but some variants have the same mobility, so that other techniques are required, such as HPLC and the use of allele-specific polymerase reaction or oligonucleotide probes. Novel variants, not previously described in Thailand, require structural analysis at the protein and DNA level. PMID- 11400797 TI - Preventation of thalassemia in Australia. AB - Screening for thalassemia and other hemoglobinopathies in the major maternity hospitals in Melbourne, Australia has shown that 6% of the patient population carries a clinically significant genetic abnormality. The most common of these are beta-thalassemia (3%). HbS (1.8%), HbE (0.5%) and alpha0 thalassemia (0.4%). Approximately 60 prenatal diagnoses for the clinically significant combinations of these abnormal genes are performed annually in the 2 major centers of Melbourne and Sydney. The majority of these prenatal diagnoses are for beta thalassemia major (65%). whilst 11% are for Bart's hydrops fetalis, 8% for HbE/beta-thalassemia. 6% for HbS/beta-thalassemia, 2% for sickle cell anemia and the remaining 8% for other combinations of thalassemia/hemoglobinopathies. Of the 178 patients with beta-thalassemia major, sickle cell disease or beta-thalassemia in combination with HbE or HbS, only 5 are less than 5 years old, reflecting both the success of the screening program and the increasing acceptance by couples of 1st trimester prenatal diagnosis. PMID- 11400798 TI - Prevalence of HB E from cord blood samples and after one year follow-up. AB - Hemoglobin (Hb) E is the most prevalent hemoglobinopathy in Southeast Asia. The prevalence of this condition varies from 9-60% of the population in different regions of Thailand and has the highest prevalence the northeast of the country. Neonatal diagnosis of Hb E can be made by detecting the Hb band in cord blood samples at the Hb A2 position using starch gel and cellulose acetate electrophoresis. Our study, performed in Bangkok, in the central part of Thailand, resealed that 182 out of 1,015 cord blood samples (17.9%) contained Hb E in amounts of between 1.9 and 10.0%. The cases who had Hb A, F and E with or without Hb Bart's were initially included in the study. These cases were suspected to have the Hb E trait. One hundred and seven cases (58.89%) were available for follow up and in all of these, Hb E could be detected throughout the study. A sharp increase in the amount of Hb E was observed at the 3 months follow-up appointment. One year follow-up could be made in 72 cases (39.6%) when the percentage of Hb E was around 25%. We conclude that measurement of Hb E in cord blood an easily accessible, simple, practical and sensitive procedure which can be used to study the Hb E hemoglobinopathy which is widely distributed in Thailand and Southeast Asia. PMID- 11400799 TI - Treatment of chronic anal fissure with topical glyceryl trinitrate: a double blind, placebo-controlled trial. AB - BACKGROUND: Topical glyceryl trinitrate (GTN) may produce healing of anal fissure by decreasing the high resting anal sphincter pressure in these patients. The present study assessed the efficacy of GTN in chronic anal fissure in a double blind placebo-controlled trial. METHOD: Patients with chronic anal fissure (for more than 8 weeks) underwent measurement of maximum anal resting pressure (MARP) before and 12 minutes after application of either 0.2% GTN or placebo ointment in a randomized manner. They then received twice-daily local application of their respective ointment for 6 weeks. Symptoms and healing of fissure were assessed; patients were evaluated at 3 months for evidence of relapse. RESULTS: 19 adult patients (12 men) were studied; 10 received GTN and 9 placebo. Mean (SD) MARP decreased from 131.0 (32.3) cm H2O to 93.5 (28.4) cm H2O (p<0.05) with GTN and from 150.5 (36.9) cm H2O to 142.8 (35.0) cm H2O (p=ns) with placebo. Fissure healed in 7 of 10 patients treated with GTN and 2 of 9 patients treated with placebo (p<0.05). There was no relapse of fissure in either group. CONCLUSION: Local application of GTN was effective in healing chronic anal fissure. PMID- 11400800 TI - Epidemiology of dyspepsia in the general population in Mumbai. AB - BACKGROUND AND AIMS: Dyspepsia is a common complaint in the general population. The prevalence, demography and economic implications of dyspepsia in India are not known; we studied these using a detailed symptom questionnaire. METHODS: 2549 presumably healthy adults (mean age 37.2 [14.1] years; 1441 men) were interviewed. Gastrointestinal symptoms, their investigation and treatment, dietary history and history of addictions were noted. Dyspepsia was defined as abdominal fullness or upper abdominal pain present for at least one month; irritable bowel syndrome (IBS) was defined by Manning's criteria. Based on the symptom profile, subjects were divided into three broad groups: no dyspepsia (n=1695; 945 men), dyspepsia with (110; 63 men) or without (664; 382 men) IBS, and IBS alone (80; 51 men). RESULTS: 774 subjects (30.4%) had dyspepsia; the median (range) duration of symptoms was 24 (1-360) months. Abdominal fullness (n=614), abdominal pain (374), heartburn (272) and belching (271) were the most common symptoms; significant symptoms (present at least once a week) occurred in 306 subjects (12.0% of the population). More than half the subjects had symptoms suggestive of mixed type of dyspepsia; dysmotility-like dyspepsia was the next most common (n=257; 33.2%). The frequency of dyspepsia was not related to type of diet or consumption of spices. Dyspepsia was more prevalent in subjects who abused tobacco or alcohol. Three hundred and twenty-one subjects with dyspepsia (41.4%) had visited a physician for their complaints and had received treatment with antacids, acid suppressors or prokinetic drugs; 4.5% and 7.2% had undergone previous endoscopy and ultrasonography, respectively; dyspeptic subjects underwent more investigations (p<0.001) than those with IBS. CONCLUSIONS: Dyspepsia is reported by almost one-third of the population in Mumbai; significant symptoms occur in 12%. Forty percent of these subjects receive treatment and only a small number undergo endoscopy or ultrasonography. PMID- 11400801 TI - Cholecystoduodenoplasty for high-output duodenal fistula. AB - External duodenal fistulae from sutured duodenal ulcer perforation sites are difficult to manage and most patients succumb to septicemia and undernutrition. This is due to failure of closure of the perforation site in the duodenum. Most techniques described in the past to facilitate closure have failed to give satisfactory results. We have devised a new procedure where the duodenal ulcer perforation is closed by mobilizing the gall bladder. A hole is made in the fundus of the gall bladder and it is anastomosed to the freshened edges of the duodenal opening. We have treated six patients by this technique. In five patients the leak was satisfactorily sealed. Three patients died - one due to persistent leak and two due to jejunostomy leak. PMID- 11400802 TI - Endocrine cell carcinoma of gall bladder. AB - Primary neuroendocrine carcinoma of the gall bladder is rare. We report a 70-year old woman with a gall bladder mass and liver metastases; fine-needle aspiration cytology from these revealed neuroendocrine carcinoma. There was no evidence of any other primary site. The patient was treated symptomatically; she died within a month of discharge from hospital. PMID- 11400803 TI - Eosinophilic gastritis--an unusual cause of gastric outlet obstruction. AB - Eosinophilic gastroenteritis is a rare entity. We report a 41-year-old man who presented with features of gastric outlet obstruction due to a submucosal lesion in the distal end of the stomach. Distal gastrectomy with Billroth II reconstruction was done. Histology showed eosinophilic gastritis infiltrating the muscular and serosal layers of the pylorus and antrum. PMID- 11400804 TI - Plasmodium falciparum and hepatitis E virus co-infection in fulminant hepatic failure. AB - Acute hepatitis E and falciparum malaria can each present with fulminant hepatic failure and are common in tropical countries. However, co-existence of these two conditions has not been reported. We report a 20-year-old girl who presented with fever and altered sensorium. Peripheral smear was positive for Plasmodium falciparum, and IgM anti-HEV was positive. She died despite antimalarial drugs and supportive management. Postmortem liver tissue showed changes suggestive of acute viral hepatitis. PMID- 11400805 TI - Laparoscopic gastropexy for chronic intermittent gastric volvulus. AB - The traditional surgical treatment of chronic gastric volvulus involves laparotomy for derotation of the stomach and its fixation to the patients. We describe a 36-year-old man with organoaxial gastric volvulus who was treated successfully with laparoscopic gastropexy. He is asymptomatic four months later. PMID- 11400806 TI - Coccidioidomycosis presenting as liver abscess. AB - A 26-year-old man presented with pain in the right upper abdomen. Ultrasonography revealed a large solitary right lobe liver abscess, which was treated by aspiration and antiamebic treatment. The patient continued to be symptomatic with the abscess increasing in size. Repeat aspiration revealed a pure growth of coccidioidomycosis, which was confirmed on culture. He was treated with amphotericin-B; the abscess resolved completely and the patient has remained asymptomatic at one-year follow up. PMID- 11400807 TI - Toxic megacolon in a renal allograft recipient with cytomegalovirus colitis. AB - We report a 35-year-old man, a renal allograft recipient, who presented with toxic megacolon. Segmental biopsies from the colon were consistent with cytomegalovirus colitis. Serum polymerase chain reaction for cytomegalovirus DNA confirmed the diagnosis. He was treated with ganciclovir but, though his abdominal condition improved initially, he worsened later and succumbed to his illness. PMID- 11400808 TI - Gastric and intestinal lactobezoars. AB - Two male full-term infants presented with unusual features of lactobezoar. One had gastric disease while the other had small bowel bezoar. The gastric lactobezoar was managed medically while the intestinal one required surgical intervention. PMID- 11400809 TI - Management of bile duct injuries--experience in a referral unit. PMID- 11400810 TI - Characteristics of gastric malignancy in eastern India. PMID- 11400811 TI - Brown Kelly-Paterson syndrome and other eponymous misnomers. PMID- 11400812 TI - Idiopathic tension pneumoperitoneum after laparotomy. PMID- 11400813 TI - A palpably enlarged hard gall bladder can be tubercular. PMID- 11400814 TI - Cryptogenic chronic liver disease in India: what is the role of hepatitis B? PMID- 11400815 TI - Infrequent occurrence of silent HBV infection among Indian patients with chronic liver disease. AB - OBJECTIVE: To investigate the occurrence of silent hepatitis B virus (HBV) infection among patients with chronic liver disease (CLD). METHODS: Plasma samples from 71 CLD patients including 9 HBsAg-positive individuals were tested for HBV DNA by nested polymerase chain reaction (nPCR), and for HBV serum markers, i.e., anti-HBc antibody, HBeAg and anti-HBe antibody. The individuals were also tested for hepatitis C virus (HCV) RNA and anti-HCV antibody. RESULTS: Among 62 HBsAg-negative patients, silent HBV infection was seen in only two (3.2%). Silent HBV infection was not found in any of the 26 patients who had evidence of HCV infection. One HBsAg-positive patient was positive for anti-HCV in the absence of HCV RNA. CONCLUSIONS: There is a low rate of silent HBV infection among patients with CLD in India, where HBV is moderately endemic. Silent HBV infection is not associated with HCV-related CLD, which is in contrast to reports from other HBV-endemic areas in Asia. PMID- 11400816 TI - Etiology and management of obscure gastrointestinal bleed--an appraisal from eastern India. AB - OBJECTIVES AND METHOD: Forty patients (mean age 45 years; 24 men) attending a tertiary care hospital in eastern India during the period 1996-2000 were investigated to evaluate the etiology and clinical spectrum of obscure gastrointestinal bleed. RESULTS: The patients presented to hospital after mean symptom duration of 2.5 years. They had received an average of 15 units of blood transfusion. Most patients presented with recurrent melena (85%); all had iron deficiency anemia. A total of 230 investigations (89 gastroscopies, 54 colonoscopies, 25 double-contrast meal and follow-through studies, 14 small bowel enemas, 24 radionuclide scans, 16 mesenteric angiographies and 8 intraoperative endoscopies) yielded positive diagnosis in 87.5% of cases. The diseases encountered were small bowel and colonic angiodysplasias (32.5%), ileal Crohn's disease (20%), intestinal tuberculosis (10%), intestinal tumors (10%), nonspecific small bowel ulcers and strictures (7.5%), Meckel's diverticulum (5%) and hemobilia (2.5%). The etiology remained obscure in 5 (12.5%) cases. Overall success of surgery was 63%; in-hospital mortality was 7.5%. CONCLUSION: Though obscure gastrointestinal bleed is commonly caused by angiodysplasias, it can be an atypical presentation of Crohn's disease. PMID- 11400817 TI - Comparison of intradermal and intramuscular administration of hepatitis B vaccine in neonates. AB - BACKGROUND: Hepatitis B virus (HBV) infection and its complications are among the most common diseases in Iran. National mass vaccination of neonates against hepatitis B was started in 1991, but was considered a costly venture. AIM: To compare the efficacy of low-dose intradermal HBV recombinant vaccine with standard intramuscular dose in neonates. METHOD: 165 apparently healthy neonates born in Shiraz were randomized to receive either 10 microg [corrected] of recombinant vaccine intramuscularly (IM; n=82) or 2 microg [corrected] vaccine intradermally (ID; n=83) at months 0, 1, 6. Anti-HBs titers were measured at 6 and 18 months after the first dose. RESULTS: 53 and 51 neonates in the IM and ID groups, respectively, completed the study. Protective anti-HBs titers (>10 IU/L) at 18 months after the first dose were achieved in 98.1% and 96.2% of neonates in IM and ID groups, respectively (p=ns). The only side effect in the ID group was local hyperpigmentation, which was seen in 55%; no significant side effect was reported in the IM group. CONCLUSION: Intradermal vaccination with 20% of standard dose is as effective as IM vaccination when evaluated at 18 months after the first dose. PMID- 11400818 TI - Induction of oxidative stress in antitubercular drug-induced hepatotoxicity. AB - BACKGROUND AND AIM: Oxidative stress could play a role in the pathogenesis of antitubercular drug (ATD)-induced hepatotoxicity. We therefore studied the plasma level of reduced glutathione (GSH) and malondialdehyde (MDA) in patients with ATD induced hepatotoxicity (cases), ATD-treated controls (disease controls) and in healthy volunteers. METHODS: This study was carried out in a case-control design. Twenty-one cases, 21 age- and sex-matched disease controls, and 10 healthy volunteers were enrolled. Plasma levels of GSH and MDA were measured. RESULTS: Plasma levels of GSH (median [range] 11.5 [6.2-21.2] mmol/dL) and MDA (1390 [560 2310] nmol/dL) of cases were significantly different (p<0.01) from GSH (18.4 [10.5-24.4]) and MDA (290 [240-550]) of disease controls. Further, plasma GSH and MDA levels of both the ATD-treated groups were different from those in healthy controls. CONCLUSION: Lower levels of plasma GSH and higher levels of MDA may be due to oxidative stress resulting from ATD therapy. PMID- 11400819 TI - Multidisciplinary care in head and neck cancer. AB - Head and neck cancer treatment is uniquely suited to multidisciplinary care. A comprehensive list of essential participants consists of a broad spectrum of health care professionals. A spirit of cooperation and mutual respect is the foundation for the synergy that results in the best possible patient care. Reconstructive surgery has emerged as a sophisticated subspecialty that fills a critical role restoring the head and neck patient to maximal health and well being. The concept of multidisciplinary care, pioneered at the nation's comprehensive cancer centers, should be adopted universally by clinicians seeking to offer the patient with head and neck cancer the best care. PMID- 11400820 TI - Prefabricated and prelaminated flaps for head and neck reconstruction. AB - Flap prefabrication and prelamination are evolving, new techniques that are useful in reconstructing complex defects of the head and neck. Flap prefabrication involves the introduction of a new blood supply by means of a vascular pedicle transfer into a volume of tissue. After a period of neovascularization, this volume of tissue may be transferred, based only on its implanted vascular pedicle. The transfer may be local transposition or by microsurgical transfer. Flap prelamination refers to a technique in which additional tissue is added to an existing flap (without manipulation of its axial blood supply) to make a multilayered flap that may be used for complex, three dimensional multilayered reconstructions. This technique may be used locally or at a distance, requiring microvascular transfer. Examples of each are described in this article. PMID- 11400821 TI - Aesthetically successful mandibular reconstruction with a single reconstruction plate. AB - Aesthetically successful mandibular reconstruction can be performed with free fibular flaps and with a single low-profile reconstruction plate. The keys to aesthetic success are accurate bending of the reconstruction plate, accurate alignment of the bone, and maintenance of the lower border of the mandible. If only the mandible and overlying oral lining are missing, the results can be indistinguishable from normal. In massive defects that include other structures besides the mandible, however, excellent aesthetic results can be difficult or impossible. Massive soft tissue deficits and heavy doses of postoperative radiation therapy can impact severely the aesthetic quality of the result. Patients should be aware of these limitations and have appropriately realistic expectations. PMID- 11400822 TI - Cheek reconstruction with laterally based flaps. AB - Laterally based cervicofacial advancement flaps that follow the borders of facial aesthetic subunits provide a more aesthetic solution to the reconstruction of anterior cheek defects than do standard cervicofacial flaps or skin grafts. The circulation in these laterally and inferiorly based flaps is improved, because the transverse facial artery is spared. These flaps also provide better contour and possess more aesthetic scar placement than do anteriorly and inferiorly based flaps. PMID- 11400823 TI - Prevention of ectropion in reconstruction of facial defects. AB - Lower eyelid malpositions and ocular damage occur with inadequate reconstructions of facial defects that encroach on the periocular region. Reconstructive principles and techniques are presented that are essential in the prevention of ectropion in these situations. Eyelid and periocular anatomy is reviewed. The use of canthalplasties, canalicular reconstruction, and ancillary techniques for facial flaps are presented. PMID- 11400824 TI - The submental flap: its uses as a pedicled or free flap for facial reconstruction. AB - The submental flap is an alternative to microsurgical flaps for facial and intraoral defects. The submental flap is simple and can be raised rapidly. It produces good color, texture, and contour match without a conspicuous site. Its versatility and its rotational arc explains its large application in facial surgery. The pedicle is reliable when the flap is not used with a reverse flow. The surgeon must know the possible presence of valves in the venous system of the face and plan to perform a venous microsurgical anastomosis if venous congestion jeopardizes the flap's survival. The location of the flap in a lymph node area restricts its uses for traumatic cases, and for benign and malignant tumors, to reduce the potential of cancer spreading to the lymph nodes (e.g., basal cell carcinoma). PMID- 11400825 TI - The use of skin grafts for nasal lining. AB - The satisfactory replacement of missing nasal lining often determines the aesthetic and functional result of a nasal reconstruction. Although full thickness skin grafts have been employed for lining in the prefabricated forehead flap technique, the results are unpredictable because they remain largely unsupported and subject to contraction. Advances in the understanding of the vasculature of the forehead flap, the use of primary and delayed cartilage grafts, and the appropriate use of operative staging permit full-thickness skin grafts to be combined with a forehead flap at the time of transfer, with little risk of loss. The technique is especially useful in the elderly or debilitated patient when a shortened operative time and less intranasal manipulation is appropriate, or when previous injury or rhinoplasty has interfered with septal blood supply, making the use of intranasal lining flaps unreliable. PMID- 11400826 TI - Close-range shotgun and rifle injuries to the face. AB - The treatment of extensive shotgun and rifle injuries to the face is extremely difficult and demands experience in microsurgery and craniofacial surgery. Early aggressive surgery with immediate bone and soft tissue reconstruction is recommended for the management of extensive facial gunshot wounds. Experience has shown that early three-dimensional bone replacement can be achieved with bone grafts in the midfacial area if the bone grafts are covered with well vascularized tissues. Large midfacial defects can be reconstructed safely and effectively with free-tissue transfers, including bone. In the acute stage, microvascular muscle flaps are preferred because of their good vascularity and good filling capacity. These flaps are able to cover the anterior cranial fossa. When vascularized bone is needed, the authors' first choice is a latissimus dorsi flap with scapular bone. Patients treated with an early and aggressive surgical strategy develop fewer problems in form of infection, contraction, scarring, and require fewer secondary corrections. Successful primary treatment allows the surgeon to use multiple modalities, including tissue expansion, free-tissue transfers, and local flaps in a noninfected environment. The treatment is rewarding, and the results are surprisingly good. It is extremely unusual for patients with self-inflicted gunshot injuries to reattempt suicide. PMID- 11400827 TI - Microsurgical reconstruction of the buccal mucosa. AB - This article addresses the reconstruction of the buccal mucosa, which is necessary after tumor resection and contracture release. PMID- 11400828 TI - Reconstruction of the hypopharynx and cervical esophagus. AB - Hypopharynx and cervical esophageal defects are challenging problems for the reconstructive surgeon. Prior surgery and radiation therapy contribute to the difficulty in managing these patients. The surgeon must possess a reconstructive algorithm that varies depending on the defect, available donor sites, and his or her experience. The free jejunal flap is the flap used for most of these defects. The radial forearm flap is reserved for partial defects measuring less than 50% of the circumference of the pharynx. The gastric pull-up is used when an intrathoracic esophagectomy is necessary. The pectoralis flap is reserved for situations when external coverage is necessary in addition to hypopharyngeal reconstruction or when a free-tissue transfer is not appropriate. Reconstruction can offer most patients successful swallowing while minimizing complications. PMID- 11400829 TI - Aesthetic facial contour reconstruction with microvascular free flaps. AB - Reconstruction of challenging surgical problems has undergone a dramatic evolution over the past quarter of a century. Microsurgical correction of facial contour deformities is a prime example of how far microsurgical reconstruction has evolved. This article discusses the current surgical techniques. PMID- 11400830 TI - The lateral arm flap. AB - The authors contend that the conventional flap (LAF) and the ELAF should be considered among the best choices of all possible fasciocutaneous flaps for head and neck reconstruction. Easy and quick dissection under tourniquet, design variability, and potential sensory innervation make these flaps superior to other fasciocutaneous flaps. The ELAF provides the same pliable, thin fasciocutaneous tissue and a longer pedicle than the RFF. In contrast to the RFF, the donor site can be closed primarily and heals with an acceptable scar without any functional deficit. The authors therefore strongly recommend clinical application of these flaps in head and neck reconstruction. PMID- 11400831 TI - Coverage of skull base defects. AB - Successful reconstruction of the cranial base requires a knowledge of this complex anatomic area, a careful assessment of the defect, a healthy respect for the potential for ascending infection and meningitis, and reliable techniques to effectively contain the intracranial space with vascularized tissue. The first step in reconstruction is a secure dural repair, which must be covered by a healthy vascularized layer. The scalp contains galeal and pericranial flaps, which are usually incorporated into the reconstruction. Sometimes, along with local muscles such as the temporalis, these local tissues are all that is needed to complete the reconstruction. When the defects are larger and in irradiated beds, free tissue transfer has emerged as the most reliable method to bolster the dural repair. PMID- 11400832 TI - Reconstruction of the voice after laryngectomy. AB - Voice reconstruction and rehabilitation are important for quality of life for patients after surgical ablation of tumors in the larynx or pharynx. In addition to the esophageal voice, the artificial larynx, and external voice devices, the following procedures have been developed: (1) after laryngectomy with preservation of pharynx, neoglottis or TEP can be performed; (2) after laryngopharyngectomy a forearm flap with TEP, or a jejunal transfer with TEP or voice tube shunt can be selected; and (3) after laryngopharyngoesophagectomy, either pharyngogastrotomy with TEP, or colon segment interposition with TEP can be employed. The voice tube shunt is improving, and allograft transplantation is currently under investigation. PMID- 11400833 TI - Mandibular reconstruction with fibula osteoseptocutaneous free flap and osseointegrated dental implants. AB - The fibula osteoseptocutaneous free flap is ideal for reconstruction of composite mandible defects. Osseointegration is possible and advantageous. Primary osseointegration of dental implants is a safe and reliable procedure in selected groups of patients, promoting early total oral rehabilitation with restoration of both function and cosmesis. Further studies are necessary to assess the specific indications for osseointegration teeth in patients with malignant tumors and in those patients who have received radiation therapy. PMID- 11400834 TI - Platysma flap for oral reconstruction. AB - In the authors' series of 12 consecutive patients who had oral reconstruction using the indications described above, flap survival always was achieved. Venous congestion was observed often: the flap became intensively red, showing a disappointing color that resolved spontaneously with only skin de epithelialization. In one case of floor-of-the-mouth repair, a marginal necrosis of the distal portion of the flap occurred, possibly because of a venous drainage problem. The secondary defect healed spontaneously, without fistula formation. The time required to outline the flap is less than for most other flaps, and donor site morbidity is minimal. In selected cases, a platysma flap is an easy and rewarding solution for repairing various oral defects. PMID- 11400835 TI - Myofibroblasts in the accessory ligament (distal check ligament) and the deep digital flexor tendon of foals. AB - OBJECTIVE: To demonstrate myofibroblasts in the accessory ligament of the deep digital flexor tendon (ie, distal check ligament) and deep digital flexor tendon of clinically normal foals. SAMPLE POPULATION: Tissue specimens from 25 foals that were necropsied for reasons unrelated to this study and unrelated to musculoskeletal disease. PROCEDURE: The distal check ligament and deep digital flexor tendon of both forelimbs were examined histologically. Myofibroblasts were identified by immunohistochemical staining specific for alpha-smooth muscle actin (alpha-SMA). RESULTS: Most of the cells in the distal check ligament and deep digital flexor tendon of all foals stained positive for alpha-SMA. CONCLUSION AND CLINICAL RELEVANCE: Myofibroblasts made up most of the cells in the distal check ligament and deep digital flexor tendon of clinically normal foals. These cells have contractile ability and therefore, may play a role in flexure contracture of these tendons. The ability of tetracycline to chelate calcium or decrease the expression of the contractile protein alpha-smooth muscle actin could inhibit the myofibroblasts' ability to contract, thus providing a rationale for tetracycline administration as a treatment of distal interphalangeal joint flexor deformity in foals. PMID- 11400836 TI - Evaluation of formalin-fixed paraffin-embedded tissues from vaccine site associated sarcomas of cats for polyomavirus DNA and antigen. AB - OBJECTIVE: To determine whether vaccine site-associated sarcomas (VSS) from cats contain polyomavirus antigen or DNA. SAMPLE POPULATION: 50 formalin-fixed paraffin-embedded tissue blocks of VSS from cats. PROCEDURE: Sections from each tissue block were evaluated for polyomavirus antigen by use of an avidin-biotin complex immunohistochemical staining method, using rabbit anti-murine polyomavirus polyclonal antiserum as the primary antibody. The DNA was extracted from sections of each tissue block, and a polymerase chain reaction assay was performed, using primers designed to amplify regions of the bovine polyomavirus genome and consensus polyomavirus primers designed to detect unknown polyomaviruses. RESULTS: Polyomavirus antigen and DNA were not detected in any of the VSS. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that polyomaviruses likely do not have any direct involvement in the pathogenesis of VSS in cats. PMID- 11400837 TI - Evaluation of formalin-fixed paraffin-embedded tissues from vaccine site associated sarcomas of cats for papillomavirus DNA and antigen. AB - OBJECTIVE: To determine whether vaccine site-associated sarcomas (VSS) from cats contain papillomavirus antigen or DNA. SAMPLE POPULATION: 50 formalin-fixed paraffin-embedded tissue blocks of VSS from cats. PROCEDURE: Sections from each tissue block were evaluated for papillomavirus antigen by use of an avidin-biotin complex immunohistochemical staining method, using rabbit anti-bovine papillomavirus type-1 antibody. The DNA was extracted from sections of each tissue block, and polymerase chain reaction assays were performed, using primers designed to amplify regions of the E5 gene of bovine papillomavirus and consensus primers designed to amplify a region of the L1 gene of animal papillomaviruses. Sections from 20 of the tissue blocks were evaluated by use of nonradioactive in situ hybridization for bovine papillomavirus DNA. RESULTS: Papillomavirus antigen and DNA were not detected in any of the VSS. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that papillomaviruses likely do not have any direct involvement in the pathogenesis of VSS in cats. PMID- 11400838 TI - Lectin binding patterns of uterine glands in mares with chronic endometrial degeneration. AB - OBJECTIVE: To evaluate changes of glycoconjugate in uterine glands of endometrial tissues obtained from mares. ANIMALS: adult mares. PROCEDURE: Uterine biopsy samples were collected during the breeding season and analyzed histologically for signs of chronic endometrial degeneration. Stage of the estrous cycle was established, using clinical examination and determination of hormonal status. Uterine tissue samples were analyzed, using lectin histochemical and immunohistochemical techniques (estrogen and progesterone receptors). Connective tissues were stained to determine alterations of ground substance in periglandular fibrosis. RESULTS: Of 50 mares, 30 (60%) were classified as normal or having modest alterations, and 20 (40%) were classified as having moderate or severe endometrial degeneration. In normal equine endometrium, several lectins (Helix pomatia agglutinin, Lotus tetragonolobus agglutinin, Ricinus communis I agglutinin, Ulex europaeus agglutinin, and wheat germ agglutinin) bound to glycoconjugates of the luminal epithelium and openings of uterine glands. Lectin binding patterns of cystic dilated glands or fibrotic glands in endometrial samples were remarkably strong, whereas normal surrounding cells remained unstained. Lotus tetragonolobus lectin was not suitable for detecting endometrial alterations. Connective tissues stained with Alcian blue and results of Hale colloidal-iron binding revealed acidic ground substance in periglandular fibrosis. Estrogen and progesterone receptors were evenly distributed in healthy and affected endometrial samples. CONCLUSIONS AND CLINICAL RELEVANCE: Glycoconjugate patterns of uterine glands were altered in mares with chronic endometrial degeneration. Therefore, uterine secretions are likely to be altered. These changes are not induced by changes in content of estrogen and progesterone receptors in endometrial tissues. PMID- 11400839 TI - Estimation of genetic population variables for six radiographic criteria of hip dysplasia in a colony of Labrador Retrievers. AB - OBJECTIVE: To estimate genetic population variables for 6 radiographic criteria of canine hip dysplasia (CHD). ANIMALS: 664 full- and half-siblings from a colony of Labrador Retrievers. PROCEDURE: Heritability estimates and genetic correlations were calculated for 6 radiographic criteria of CHD. Two evaluation protocols were compared: the grade of the most severely affected hip joint and the sum of the scores for both hip joints. The predictive performance of estimated breeding values was also evaluated. RESULTS: The overall prevalence of CHD (Federation Cynologique Internationale grades C, D, and E) was 29.6%. Median age at radiographic examination was 377 days. Heritability for the total CHD grade, Norberg angle (NA), coverage of the femoral head (COV), craniodorsal acetabular rim (ACR), subchondral bone sclerosis (SUBCH), shape of the femoral head and neck (FHN), and osteoarthritic changes at the insertion site of the joint capsule (JC) was estimated as follows: 0.44, 0.43, 0.46, 0.37, 0.32, 0.21, and 0.05, respectively. Heritability estimates were slightly higher for the sum of the scores for both hip joints. If NA and COV were included as fixed effects in the model for the dependent variables ACR, SUBCH, FHN, and JC , then heritability of these traits significantly decreased (0.08 to 0.15). High scores of NA and COV lead to a significant increase of the scores of the remaining criteria. CONCLUSIONS AND CLINICAL RELEVANCE: Canine hip dysplasia is heritable to a moderate degree. Signs of subluxation revealed the highest heritability estimates. The criteria ACR, SUBCH, FHN, and JC were strongly influenced by NA and COV. PMID- 11400840 TI - Fluorophotometric determination of aqueous humor flow rate in clinically normal dogs. AB - OBJECTIVE: To determine aqueous humor flow rate in clinically normal dogs, using fluorophotometry. ANIMALS: 20 clinically normal Beagles. PROCEDURE: A study was performed on 5 dogs to establish an optimal protocol for fluorophotometric determination of aqueous humor flow rate. This protocol then was used to measure aqueous humor flow rate in 15 dogs. Corneas were loaded with fluorescein by topical application, and corneal and aqueous humor fluorescein concentrations were measured 5, 6.5, and 8 hours after application. Concentration-versus-time plots were generated, and slopes and ratios of the fluorescein concentration in the cornea and aqueous humor from these graphs were used to calculate flow rates. Calculations were performed by use of automated software provided with the fluorophotometer and by manual computation, and the 2 calculation methods were compared. RESULTS: The protocol established for the 5 dogs resulted in semilogarithmic and parallel decay of corneal and aqueous humor concentrations. Manually calculated mean +/- SD aqueous humor flow rates for left, right, and both eyes were 5.58 +/- 2.42, 4.86 +/- 2.49, and 5.22 +/- 1.87 microl/min, respectively, whereas corresponding flow rates calculated by use of the automated software were 4.54 +/- 3.08, 4.54 +/- 3.10, and 4.54 +/- 2.57 microl/min, respectively. Values for the left eye were significantly different between the 2 computation methods. CONCLUSIONS AND CLINICAL RELEVANCE: Aqueous humor flow rates can be determined in dogs, using fluorophotometry. This technique can be used to assess pathologic states and medical and surgical treatments that alter aqueous humor dynamics. PMID- 11400841 TI - Effects of topical application of a 2% solution of dorzolamide on intraocular pressure and aqueous humor flow rate in clinically normal dogs. AB - OBJECTIVE: To evaluate effects of topical application of a 2% solution of dorzolamide on intraocular pressure (IOP) and aqueous humor flow rate in clinically normal dogs. ANIMALS: 15 Beagles. PROCEDURE: The IOP was measured in both eyes of all dogs for 3 days to determine baseline values. In a single-dose study, 50 microl of dorzolamide or control solution was applied in both eyes at 7:00 AM, and IOP was measured 7 times/d. In a multiple-dose study, dorzolamide or control solution was applied to both eyes 3 times/d for 6 days, and IOP was measured 4 times/d during treatment and for 5 days after cessation of treatment. Aqueous humor flow rate was measured for all dogs fluorophotometrically prior to treatment and during the multiple-dose study. RESULTS: In the single-dose study, dorzolamide significantly decreased IOP from 30 minutes to 6 hours after treatment. Mean decrease in IOP during this time span was 3.1 mm Hg (18.2%). Maximal decrease was detected 6 hours after treatment (3.8 mm Hg, 22.5%). In the multiple-dose study, dorzolamide decreased IOP at all time points, and maximal decrease was detected 3 hours after treatment (4.1 mm Hg, 24.3%). Mean aqueous humor flow rate decreased from 5.9 to 3.4 microl/min (43%) after treatment in the dorzolamide group. CONCLUSIONS AND CLINICAL RELEVANCE: Topical application of a 2% solution of dorzolamide significantly decreases IOP and aqueous humor flow rate in clinically normal dogs. Therefore, topical administration of dorzolamide should be considered for the medical management of dogs with glaucoma. PMID- 11400842 TI - Molecular epidemiologic features and antimicrobial susceptibility profiles of various ribotypes of Pseudomonas aeruginosa isolated from humans and ruminants. AB - OBJECTIVES: To assess automated ribotyping for characterization of Pseudomonas aeruginosa isolates and to identify their type prevalence and geographic distribution. SAMPLE POPULATION: 39 human and 56 ruminant P aeruginosa isolates. PROCEDURES: Isolates were identified by use of bacteriologic techniques and automated Pvull-based ribotyping. Susceptibility to antimicrobials was tested in vitro. Data were analyzed for index of discrimination; prevalence ratio; geographic distribution of ribotypes found only in humans, only in cows, or only in goats (single-host ribotypes); and geographic distribution of ribotypes found in humans and ruminants (multihost ribotypes). RESULTS: All isolates were typeable (45 ribotypes, 35 single-host ribotypes). Ribotyping index of discrimination was 0.976. More isolates (45.3%) than expected yielded multihost ribotypes (22% of all ribotypes). Although 8.6% of single-host ribotypes were found in 4 or more isolates, 60% of multihost ribotypes were found in 4 or more isolates. Ninety percent of multihost ribotypes were isolated from different geographic areas, whereas 3.0% of single-host ribotypes were isolated from different geographic areas. All ruminant isolates were susceptible to gentamicin and polymyxin B. In contrast, antibiogram profiles differed for human isolates from different geographic areas. Susceptibility to antimicrobials differentiated 6 isolates not distinguished by ribotyping. CONCLUSIONS AND CLINICAL RELEVANCE: Automated ribotyping with Pvull discriminated more isolates than in vitro antimicrobial susceptibility. In combination, both tests provided more information than either test alone. Given the greater prevalence and geographic distribution of multihost ribotypes, immunocompromised humans and lactating ruminants may have a greater risk for disease if exposed to multihost P aeruginosa ribotypes, compared with single-host ribotypes. PMID- 11400843 TI - A rapid and strong laparoscopic-assisted gastropexy in dogs. AB - OBJECTIVE: To develop a technique for laparoscopic gastropexy in dogs and evaluate effects on stomach position and strength of the adhesion between the stomach and abdominal wall. ANIMALS: 8 healthy dogs. PROCEDURE: Dogs were anesthetized, and the abdomen was insufflated with carbon dioxide. A laparoscope was placed through a cannula inserted on the abdominal midline caudal to the umbilicus. Babcock forceps placed through a cannula inserted lateral to the right margin of the rectus abdominus muscle were used to exteriorize the pyloric antrum, a longitudinal incision was made through the serosa and muscular layer of the pyloric antrum, and the seromuscular layer of the pyloric antrum was sutured to the transversus abdominus muscle. After surgery, positive-contrast gastrography was used to evaluate stomach position and the onset of gastric emptying, and ultrasonography was used to assess stomach wall activity and mobility. Dogs were euthanatized 1 month after surgery, and tensile strength of the adhesion was tested. RESULTS: In all dogs, stomach position and the onset of gastric emptying were normal 25 days after surgery, and the pyloric antrum was firmly attached to the abdominal wall 30 days after surgery. Mean +/- SD ultimate load of the adhesion in tension was 106.5 +/- 45.6 N. CONCLUSIONS AND CLINICAL RELEVANCE: The laparoscopic gastropexy technique described in the present study could be performed quickly and easily by an experienced surgeon, resulted in a strong fibrous adhesion between the stomach and abdominal wall, and appeared to cause minimal stress to the dogs. PMID- 11400844 TI - Biochemical characterization of cartilage affected by osteochondritis dissecans in the humeral head of dogs. AB - OBJECTIVE: To determine glycosaminoglycan (GAG) concentration and immunohistochemical staining characteristics of type-I, -II, and -X collagen from cartilage affected by osteochondritis dissecans (OCD) in dogs. ANIMALS: 31 dogs with OCD and 11 clinically normal purpose-bred dogs. PROCEDURE: Cartilage samples were evaluated microscopically, and GAG content was determined. Immunohistochemical staining was performed for type-I, -II, and -X collagen. Sections were subjectively evaluated for location and intensity of staining. RESULTS: Cartilage affected by OCD had a variety of pathologic changes and significantly lower GAG concentrations than did normal cartilage. Normal cartilage had no detectable type-I collagen. For dogs < 9 months of age, cartilage affected by OCD had significantly more type-I collagen but significantly less type-X collagen than did control cartilage. For dogs > 12 months of age, cartilage affected by OCD contained significantly more type-I collagen than did control cartilage. There was a significant negative correlation between immunoreactivity of type-I collagen and that of type-II and -X collagen. A significant positive correlation was found between immunoreactivity of type-II and -X collagen. CONCLUSIONS AND CLINICAL RELEVANCE: Cartilage affected by OCD contains less GAG, more type-I collagen, and less type-X collagen, compared with normal cartilage. A direct correlation between these changes and the etiopathogenesis of OCD was not established. PMID- 11400845 TI - Safety and efficacy of preoperative administration of meloxicam, compared with that of ketoprofen and butorphanol in dogs undergoing abdominal surgery. AB - OBJECTIVE: To compare the safety and efficacy of preoperative administration of meloxicam with that of ketoprofen and butorphanol in dogs undergoing abdominal surgery. ANIMALS: 36 dogs undergoing laparotomy, splenectomy, or cystotomy. PROCEDURE: Dogs were randomly assigned to 1 of 3 groups. In the first part of the study, dogs were given a single dose of meloxicam, ketoprofen, or a placebo, and buccal mucosal bleeding times were measured. In the second part of the study, dogs were given meloxicam, ketoprofen, or butorphanol prior to surgery. Dogs in the butorphanol group received a second dose immediately after surgery. Pain scores (1 to 10) were assigned hourly for 20 hours after surgery and used to determine an overall efficacy score for each dog. Dogs with a pain score > or =3 were given oxymorphone for pain. Dogs were euthanatized 8 days after surgery, and gross and histologic examinations of the liver, kidneys, and gastrointestinal tract were conducted. RESULTS: Overall efficacy was rated as good or excellent in 9 of the 12 dogs that received meloxicam, compared with 9 of the 12 dogs that received ketoprofen and only 1 of the 12 dogs that received butorphanol. No clinically important hematologic, biochemical, or pathologic abnormalities were detected. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that preoperative administration of meloxicam is a safe and effective method of controlling postoperative pain for 20 hours in dogs undergoing abdominal surgery; the analgesic effects of meloxicam were comparable to those of ketoprofen and superior to those of butorphanol. PMID- 11400846 TI - Metabolic and structural abnormalities in dogs with early left ventricular dysfunction induced by incessant tachycardia. AB - OBJECTIVE: To assess morphologic and metabolic abnormalities in dogs with early left ventricular dysfunction (ELVD) induced by rapid right ventricular pacing (RRVP). ANIMALS: 7 Beagles. PROCEDURE: Plasma carnitine concentrations were measured before and after development of ELVD induced by RRVP. At the same times, transvenous endomyocardial biopsy was performed, and specimens were submitted for determination of myocardial carnitine concentrations and histologic, morphometric, and ultrastructural examination. RESULTS: In 4 dogs in which baseline plasma total carnitine concentration was normal, RRVP induced a decrease in myocardial total and free carnitine concentrations and an increase in myocardial esterified carnitine concentration. In 3 dogs in which baseline plasma total carnitine concentration was low, plasma and myocardial carnitine concentrations were unchanged after pacing. Structural changes associated with pacing included perinuclear vacuolization in 3 dogs. Morphometric analyses indicated there was a decrease in myofiber cross-sectional diameter and area following pacing. Electron microscopy revealed changes in myofibrils and mitochondria following pacing. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that moderate to severe alterations in myocyte cytoarchitecture are present in dogs with ELVD induced by RRVP and that in dogs with normal plasma carnitine concentrations, myocardial carnitine deficiency may be a biochemical marker of ELVD. Results also indicated that transvenous endomyocardial biopsy can be used to evaluate biochemical and structural myocardial changes in dogs with cardiac disease. PMID- 11400847 TI - Effects of ground surface deformability, trimming, and shoeing on quasistatic hoof loading patterns in horses. AB - OBJECTIVE: To determine whether solar load distribution pattern on a solid nondeformable ground surface is the product of contact erosion and is the mirror image of load distribution on a deformable surface in horses. ANIMALS: 30 clinically normal horses. PROCEDURES: Solar load distribution was compared among 25 clinically normal horses during quasistatic loading on a solid nondeformable surface and on a highly deformable surface. Changes in solar load distribution patterns were evaluated in 5 previously pasture-maintained horses housed on a flat nondeformable surface. Changes in solar load distribution created by traditional trimming and shoeing were recorded. RESULTS: Unshod untrimmed horses had a 4-point (12/25, 48%) or a 3-point (13/25, 52%) wall load distribution pattern on a flat solid surface. Load distribution on a deformable ground surface was principally solar and located transversely across the central region of the foot. Ground surface contact areas on solid (24.2 +/- 8.62 cm2) and deformable (69.4 +/- 22.55 cm2) surfaces were significantly different. Maintaining unshod horses on a flat nondeformable surface resulted in a loss of the 3- and 4-point loading pattern and an increase in ground surface contact area (17.9 +/- 2.77 to 39.9 +/- 12.77 cm2). Trimming increased ground surface contact area (24.2 +/- 8.60 to 45.7 +/- 14.89 cm2). CONCLUSION AND CLINICAL RELEVANCE: In horses, the solar surface is the primary weight-loading surface, and deformability of ground surface may have a role in foot expansion during loading. Increased surface area induced by loading on deformable surfaces, trimming, and shoeing protects the foot. PMID- 11400848 TI - Effect of walking velocity on ground reaction force variables in the hind limb of clinically normal horses. AB - OBJECTIVE: To measure the effect of subject velocity on hind limb ground reaction force variables at the walk and to use the data to predict the force variables at different walking velocities in horses. ANIMALS: 5 clinically normal horses. PROCEDURE: Kinematic and force data were collected simultaneously. Each horse was led over a force plate at a range of walking velocities. Stance duration and force data were recorded for the right hind limb. To avoid the effect of horse size on the outcome variables, the 8 force variables were standardized to body mass and height at the shoulders. Velocity was standardized to height at the shoulders and expressed as velocity in dimensionless units (VDU). Stance duration was also expressed in dimensionless units (SDU). Simple regression analysis was performed, using stance duration and force variables as dependent variables and VDU as the independent variable. RESULTS: Fifty-six trials were recorded with velocities ranging from 0.24 to 0.45 VDU (0.90 to 1.72 m/s). Simple regression models between measured variables and VDU were significant (R2 > 0.69) for SDU, first peak of vertical force, dip between the 2 vertical force peaks, vertical impulse, and timing of second peak of vertical force. CONCLUSION AND CLINICAL RELEVANCE: Subject velocity affects vertical force components only. In the future, differences between the forces measured in lame horses and the expected forces calculated for the same velocity will be studied to determine whether the equations can be used as diagnostic criteria. PMID- 11400849 TI - Use of a murine xenograft model for canine transmissible venereal tumor. AB - OBJECTIVE: To develop a murine model for canine transmissible venereal tumor (CTVT). ANIMALS: Thirty-three 6-week-old NOD/LtSz-scid (NOD/SCID) mice and seven 6-week-old C57BL/6J mice. PROCEDURE: Samples of CTVT were excised from a 3-year old dog and inoculated SC into ten 6-week-old NOD/SCID mice to induce growth of xenograft transmissible venereal tumor (XTVT). To establish mouse-to-mouse transmission, samples of XTVT were removed and inoculated SC into 4 groups of 6 week-old NOD/SCID mice and into a control group. Samples of CTVT were also inoculated into immunocompetent C57BL/6J mice for a mouse antibody production (MAP) test. The canine and xenografted tumors were evaluated cytologically and histologically, and polymerase chain reaction was performed for detection of the rearranged LINE/c-MYC junction. RESULTS: 8 of 10 NOD/SCID mice that were inoculated with CTVT developed tumors 3 to 10 weeks after inoculation. In the second-generation xenograft, all mice developed tumors by postinoculation day 47; 1 X 10(6) of XTVT cells were enough to create a xenograft. Metastases developed in 4 of 20 mice. Xenografted and metastatic tumors retained cytologic, histologic, and molecular characteristics of CTVT. Results of the MAP test were negative for all pathogens. CONCLUSION: We established an NOD/SCID murine model for XTVT and metastasis of CTVT. This model should facilitate study of tumor transplantation, progression, and metastasis and should decrease or eliminate the need for maintaining allogenic transfer in dogs. PMID- 11400850 TI - Nitrogen balance in clinically normal dogs receiving parenteral nutrition solutions. AB - OBJECTIVE: To determine nitrogen balance in clinically normal dogs receiving parenteral nutrition solutions. ANIMALS: 8 clinically normal female Beagles. PROCEDURE: Dogs were randomly assigned to receive 4 treatments in random order. Treatment A consisted of IV administration of nonlactated Ringer's solution. Treatments B, C, and D consisted of IV administration of isocaloric parenteral solutions containing 0, 1.36, and 2.04 g of amino acids/kg of body weight/d, respectively, for 7 consecutive days. Urine and feces were collected on days 5, 6, and 7 of each treatment period, and Kjeldahl analysis was used to determine nitrogen balance. RESULTS: Mean nitrogen balance was negative with treatments A and B but was not significantly different from 0 with treatments C and D. Dogs had the lowest nitrogen balance values and lost the most weight while receiving treatment A. Dogs were able to conserve protein and had higher nitrogen balance values when receiving treatment B, compared with treatment A. Dogs lost the least amount of weight while receiving treatment D. Regression analysis indicated that an IV amino acid intake of 2.32 g/kg/d (95% confidence interval, 2.00 to 2.81 g/kg/d), as supplied by the commercial product used in this study, would result in zero nitrogen balance in clinically normal dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that IV amino acid requirement of clinically normal dogs is approximately 2.3 g/kg/d. PMID- 11400851 TI - Effects of oral vitamin E supplementation during late gestation in beef cattle that calved in late winter and late summer. AB - OBJECTIVE: To determine effects of breed and oral vitamin E supplementation during late gestation on serum vitamin E and IgG concentrations in beef cows that calved in late winter and late summer and in neonatal calves. ANIMALS: 73 Angus and 43 Hereford primiparous and multiparous cows and their calves. PROCEDURE: Cows in groups that were homogeneous regarding breed and age distribution were randomly allotted to groups that were orally supplemented (n = 59) or not supplemented (57) with vitamin E beginning 30 days prior to onset of 65-day calving seasons. Supplemental vitamin E was provided in a vitamin-mineral mix offered free-choice until parturition. RESULTS: Cows that calved in late winter and were supplemented orally with vitamin E had higher serum vitamin E concentrations at calving and after calving than did unsupplemented cows; differences between groups before calving were not significant. Calves from supplemented multiparous cows had higher vitamin E concentrations than did calves from unsupplemented cows. Winter-born calves from supplemented Hereford cows had heavier 205-day adjusted weaning weights than did winter-born calves from unsupplemented Hereford cows. Supplementation did not affect vitamin E or IgG concentrations in the herd that calved in late summer and did not affect calf growth. CONCLUSIONS AND CLINICAL RELEVANCE: Oral vitamin E supplementation during late gestation may be economically beneficial in certain cow-calf operations in which late-gestation cows are consuming stored forages. PMID- 11400852 TI - Morphometric and anatomic study of the hind limb of a dog. AB - OBJECTIVE: To obtain the anatomic and morphometric data required for biomechanical analysis of the hind limb in dogs. ANIMALS: A healthy adult mixed breed 23-kg male dog. PROCEDURE: Following euthanasia of the dog, all muscles of the right hind limb were identified and meticulously removed. Physiologic cross sectional areas (PCSA) and architectural indices (AI) were calculated. The coordinates for the origin and insertion of each muscle were determined, using orthogonal right-handed coordinate systems embedded in the pelvis, femur, and tibia. RESULTS: PCSA and AI were calculated for 29 muscles, and coordinates for the origins and insertions of these muscles were determined. CONCLUSIONS: Results provide the morphometric and anatomic data necessary for 3-dimensional biomechanical studies of the hind limb in dogs. PMID- 11400853 TI - Selenium status of cats in four regions of the world and comparison with reported incidence of hyperthyroidism in cats in those regions. AB - OBJECTIVE: To assess selenium (Se) status of cats in 4 regions of the world and to compare results for Se status with reported incidence of hyperthyroidism in cats in those regions. ANIMALS: 50 cats (30 from 2 regions with an allegedly high incidence of hyperthyroidism and 20 from 2 regions in which the disease is less commonly reported). PROCEDURE: Hematologic samples (heparinized whole blood, plasma, and RBC fractions) were obtained from 43 healthy euthyroid cats and 7 hyperthyroid cats. Plasma concentration of Se and activity of glutathione peroxidase (GPX) in whole blood and plasma were determined. RESULTS: Plasma concentration of Se and GPX activity in whole blood or plasma did not differ significantly among cats from the 4 regions. However, cats had a plasma concentration of Se that was approximately 5 times the concentration reported in rats and humans. The GPX activity in whole blood or plasma in cats generally was higher than values reported in rats or humans. CONCLUSIONS AND CLINICAL RELEVANCE: Cats have higher Se concentrations in plasma, compared with values for other species. However, Se status alone does not appear to affect the incidence of hyperthyroidism in cats. High Se concentrations may have implications for health of cats if such concentrations are influenced by the amount of that micronutrient included in diets. PMID- 11400854 TI - Comparison of serum parathyroid hormone and ionized calcium and magnesium concentrations and fractional urinary clearance of calcium and phosphorus in healthy horses and horses with enterocolitis. AB - OBJECTIVE: To evaluate calcium balance and parathyroid gland function in healthy horses and horses with enterocolitis and compare results of an immunochemiluminometric assay (ICMA) with those of an immunoradiometric assay (IRMA) for determination of serum intact parathyroid hormone (PTH) concentrations in horses. ANIMALS: 64 horses with enterocolitis and 62 healthy horses. PROCEDURES: Blood and urine samples were collected for determination of serum total calcium, ionized calcium (Ca2+) and magnesium (Mg2+), phosphorus, BUN, total protein, creatinine, albumin, and PTH concentrations, venous blood gases, and fractional urinary clearance of calcium (FCa) and phosphorus (FP). Serum concentrations of PTH were measured in 40 horses by use of both the IRMA and ICMA. RESULTS: Most (48/64; 75%) horses with enterocolitis had decreased serum total calcium, Ca2+, and Mg2+ concentrations and increased phosphorus concentrations, compared with healthy horses. Serum PTH concentration was increased in most (36/51; 70.6%) horses with hypocalcemia. In addition, FCa was significantly decreased and FP significantly increased in horses with enterocolitis, compared with healthy horses. Results of ICMA were in agreement with results of IRMA. CONCLUSIONS AND CLINICAL RELEVANCE: Enterocolitis in horses is often associated with hypocalcemia; 79.7% of affected horses had ionized hypocalcemia. Because FCa was low, it is unlikely that renal calcium loss was the cause of hypocalcemia. Serum PTH concentrations varied in horses with enterocolitis and concomitant hypocalcemia. However, we believe low PTH concentration in some hypocalcemic horses may be the result of impaired parathyroid gland function. PMID- 11400855 TI - Evaluation of the survival prediction index as a model of risk stratification for clinical research in dogs admitted to intensive care units at four locations. AB - OBJECTIVE: To prospectively evaluate a survival prediction index (SPI) in dogs admitted to intensive care units (ICU) and to generate and test an improved SPI (ie, SPI2). SAMPLE POPULATION: Medical records of 624 critically ill dogs admitted to an ICU. PROCEDURE: Data were collected from dogs within 24 hours after admission to an ICU. Variables recorded reflected function of vital organ systems, severity of underlying physiologic derangement, and extent of physiologic reserve; outcome was defined as dogs that survived or did not survive until 30 days after admission to the ICU. Probabilities of survival were calculated, using an established model (SPI). We then performed another logistic regression analysis, thereby reestimating the variables to create the new SPI2. Cross-validation of the models obtained was performed by randomly assigning the total sample of 624 dogs into an estimation group of 499 dogs and validation group of 125 dogs. RESULTS: Testing of SPI resulted in an area under the curve (AUC) of 0.723. Testing of SPI2 revealed an AUC of 0.773. A backwards-elimination procedure was used to create a model containing fewer variables, and variables were sequentially eliminated. The AUC for the reduced model of SPI2 was 0.76, indicating little loss in predictive accuracy. CONCLUSIONS AND CLINICAL RELEVANCE: The new SPI2 objectively stratified clinical patients into groups according to severity of disease. This index could provide an important tool for clinical research. PMID- 11400856 TI - Evaluation of a tapered-sleeve transcortical pin to reduce stress at the bone-pin interface in metacarpal bones obtained from horses. AB - OBJECTIVE: To evaluate stiffness and bone-pin interface stress for a transcortical tapered-sleeve pin (TSP) that incorporates bilateral tapered sleeves over a transcortical pin. SAMPLE POPULATION: 14 third metacarpal bones (MCIII) collected from adult horses of various breeds. PROCEDURE: Each MCIII was cut in half to provide 2 test specimens. Pins (conventional and TSP) of 3 diameters (6.35, 7.94, and 9.50 mm) were inserted in specimens (3 specimens for each diameter and each type of pin). The test fixture simulated a typical sidebar span skeletal fixation device for horses. Single cycle load-deflection tests were performed. Cyclic fatigue tests of TSP were performed to evaluate fatigue characteristics and stress conditions at the bone-pin interface. Maximum stress and strain were calculated, and results were compared with existing data on fatigue characteristics of bone. RESULTS: Significant increases in stiffness (load-deflection) and higher loads at yield point were detected for the TSP (stiffness for conventional 9.50mm pins, 4,500 N/mm; stiffness for TSP, 19,988 N/mm). Results of cyclic tests revealed a close correlation with existing data on fatigue characteristics. CONCLUSIONS AND CLINICAL RELEVANCE: The TSP described here is stiffer than conventional transcortical pins, and stress across the bone pin interface is more evenly distributed. Use of this TSP should minimize major problems encountered during external fixation associated with the transcortical pin and bone-pin interface (ie, bone necrosis, infection of the pin track, pin loosening, and bone failure). PMID- 11400857 TI - Effects of dilated cardiomyopathy on the renin-angiotensin-aldosterone system, atrial natriuretic peptide activity, and thyroid hormone concentrations in dogs. AB - OBJECTIVE: To evaluate the effect of dilated cardiomyopathy (DCM) on activity of the renin-angiotensin-aldosterone system (RAAS), the N-terminal fragment of proatrial natriuretic peptide (NT-proANP), and thyroid hormone concentrations in dogs. ANIMALS: 15 dogs with clinical signs of DCM, 15 dogs without clinical signs of DCM, and 15 age-, breed-, and sex-matched control dogs. PROCEDURE: Physical examinations, thoracic radiography, ECG, and echocardiography were performed on all dogs, and blood and urine samples were collected. RESULTS: Plasma renin activity (PRA), plasma aldosterone concentration (PAC), urine aldosterone-to creatinine ratio, and NT-proANP concentrations were significantly increased in dogs with clinical signs of DCM, compared with dogs without clinical signs and control dogs. Thyroid-stimulating hormone and total thyroxine concentrations did not differ significantly among groups; however, free thyroxine (FT4) concentrations were significantly decreased in dogs with clinical signs of DCM, compared with control dogs and DCM-dogs without clinical signs. Concentrations of PRA, PAC, FT4, and urine aldosterone-to-creatinine ratio were significantly correlated, whereas plasma concentrations of NT-proANP only correlated with FT4 concentration. CONCLUSION AND CLINICAL RELEVANCE: In dogs with clinical signs of DCM, increased concentrations of components of the RAAS were associated with increased concentrations of NT-proANP Analysis of the neurohormonal system may aid in identification of clinical stages of DCM for groups of dogs, but the range is too great and there are too many dogs that have neurohormonal concentrations within reference ranges to assess dogs on an individual basis. PMID- 11400858 TI - Comparison of the radius of curvature of the ulnar trochlear notch of Rottweilers and Greyhounds. AB - OBJECTIVE: To compare radius of curvature along the ulnar trochlear notch of Rottweilers and Greyhounds to determine whether morphologic differences exist that may contribute to the cause and pathogenesis of fragmented coronoid process in Rottweilers. SAMPLE POPULATION: Paired elbow joints from 13 Rottweilers and 14 Greyhounds. PROCEDURE: Elbow joints were radiographically scored on the basis of severity of osteoarthritic lesions. The articular contour of each ulnar trochlear notch was digitized. The radius of curvature at defined points along the ulnar trochlear notch was compared between breeds. RESULTS: Radius of curvature of the ulnar trochlear notch was not a constant function of arc length in either breed but had a consistent characteristic appearance in both breeds. Radius of curvature was greatest at each end of the ulnar trochlear notch and had 2 peaks in the midportion of the notch in both breeds. These peaks occurred farther distally in the notch and were larger in Rottweiler ulnae than Greyhound ulnae. A significant difference in mean radius of curvature was detected between breeds at these peaks. Greyhounds had significantly greater mean radius of curvature at the end of the medial coronoid process, compared with Rottweilers. CONCLUSIONS AND CLINICAL RELEVANCE: Radius of curvature of the ulnar trochlear notch is a complex function of arc length in Rottweilers and Greyhounds. The waveform has a consistent characteristic appearance in both breeds. Although significant differences were identified between breeds, associations between these differences and cause or pathogenesis of fragmented coronoid process in Rottweilers were not apparent. PMID- 11400859 TI - Expression and distribution of the Kit receptor in bovine bone marrow cells. AB - OBJECTIVE: To characterize the expression and distribution of the Kit receptor in bovine bone marrow cells (BMC) and to define the function of its ligand, stem cell factor (SCF). ANIMALS: Six 7- to 70-day-old healthy male Holstein-Friesian calves. PROCEDURES: Expression and distribution of the Kit receptor were assessed by use of flow cytometry with monoclonal antibodies (mAb) against the bovine Kit protein. Using Giemsa-stained centrifuged preparations, the histologic appearance of Kit receptor positive (Kit+) BMC were evaluated. Semisolid cultures supplemented with granulocyte colony-stimulating factor (G-CSF) and SCF were used to measure the colony formation capacity of Kit+ BMC. RESULTS: The Kit receptor was expressed on approximately 18% of total BMC. Most of Kit+ BMC did not coexpress lineage markers, but a small subset of this population did coexpress CD3. The Kit+CD3- BMC were a heterogeneous cell population comprising blast-like cells such as myeloblasts, promyelocytes, rubriblasts, and prorubricytes. Conversely, Kit+CD3+ BMC had a lymphocyte-like appearance. Kit+ BMC formed colonies in semisolid culture with G-CSF, whereas Kit- BMC failed to grow. Addition of SCF to G-CSF resulted in superadditive enhancement in colony numbers and size. CONCLUSIONS: The Kit receptor is expressed primarily on immature blood cells in bovine bone marrow, and Kit+ BMC contain hematopoietic progenitor cells that are reactive to G-CSF. In addition, SCF synergizes with G-CSF to stimulate colony formation by these cells. Our results suggest that the Kit receptor and its ligand, SCF, are involved in early stages of granulopoiesis in calves. PMID- 11400860 TI - Leukotriene metabolism by intrapulmonary vessels of newborn lambs: effect of platelet-activating factor. AB - Leukotrienes (LTs) are potent vasoconstrictors in the pulmonary circulation. We investigated LTB4 and LTE4 metabolism by intrapulmonary arteries and veins of 2 to 9 days old lambs (n = 6). Paired vessels were incubated under baseline, and stimulated conditions. LTB4 and LTE4 were extracted from media, quantfied by enzyme-linked immunosorbent assay (ELISA), normalized to tissue weight and presented as ng/mg tissue (means +/- SEMs). In arteries, baseline synthesis of LTB4 was 0.15+/-0.20 and increased to 0.96+/-0.04 on stimulation with 1.0 micromol/L A2318, and 1.74+/-0.25 with 0.1 mmol/L arachidonic acid (AA). In veins the corresponding values were 0.28+/-0.10, 2.50+/-0.51, and 5.36+/-0.70. Baseline production of LTB4 was higher in veins. LTE4 synthesis in arteries was 0.25+/ 0.02, which increased to 0.42+/-0.05 with A23187, and further to 0.69+/-0.06 with AA. The corresponding values in veins were 0.23+/-0.05, 0.74+/-0.09, and 1.56+/ 0.28. Baseline metabolism of LTE4 by the vessels was not different. Furthermore, stimulation of vessels with 50 nmol/L PAF led to over 3-fold increase in LTB4 and LTE4 metabolism by the vessels. Smooth muscle cells stimulated with A23187 metabolized LTB4 and LTC4, which was sequentially catabolized to LTD4 and LTE4. Generally, stimulated veins, whether vessels or smooth muscle cells, metabolized more leukotrienes. The selective 5-lipoxygenase inhibitor, AA-861, significantly attenuated synthesis of both leukotrienes. Western analysis of membrane protein showed gReater expression of 5-lipoxygenase in stimulated veins. Our data show that veins produce more leukotrienes due to greater expression of 5-lipoxygenase in the vessels, and suggest that veins of newborn lamb lungs may be more susceptible to LT-induced vascular reactivity in the pulmonary circulation. PMID- 11400861 TI - Metabolism of surfactant phosphatidylcholine molecular species in cftr(tm1HGU/tm1HGU) mice compared to MF-1 mice. AB - In cftr(tmIHGU/m1HGU) mice, an animal model designed to study pathophysiologic alterations due to the CFTR defect found in cysticfibrosis, surfactant phospholipids of bronchoalveolar lavage fluid (BALF) are increased. To study the metabolical basis of such increases, we intraperitoneally injected cft(tm1HGU/tm1HGU) mice [methyl-3H]choline and measured [methyl-3H]choline incorporation into phosphatidylcholine (PC) molecular species of lung tissue and BALF after 1.5 to 24 hours. MF1 and MF1 x cftr(tm1HGU/tm1HGU) hybrid mice served as controls. In tissue [methyl-3H]choline incorporation into total PC was constant for 24 hours and identical in control and cftr(tmIHGU/m1HGU) mice. However, from 7.5 to 24 hours there was a shift of [methyl-3H]choline incorporation from palmitoyloleoyl-PC and palmitoyllinoleoyl-PC towards PC species enriched in surfactant, dipalmitoyl-PC, palmitoylmyristoyl-PC, and palmitoylpalmitoleoyl-PC. The relative and absolute 3H-labels of PC species were identical for cftr(tmIHGU/m1HGU) compared to control mice. In BALF [methyl 3H]choline of total PC increased from 1.5 to 24 hours (R2 > .98), mainly due to [methyl-3H]choline-labelled dipalmitoyl-PC, in all experimental groups. In BALF from cftr(tmIHGU/m1HGU) mice, the [methyl-3H]choline label of total PC and individual PC species was significantly increased over control values after 24 hours, but not after 1.5 to 6 hours. Numbers and composition of BALF cells were not different between controls and cftr(tmIHGU/m1HGU) mice. We, conclude that increased alveolar phospholipid in cftr(tmIHGU/m1HGU) mice is likely due to decreased reuptake of surfactant. PMID- 11400862 TI - Comparison of clearance of particles inhaled with bolus and extremely slow inhalation techniques. AB - Ten healthy nonsmokers inhaled 6-microm (aerodynamic diameter) Teflon particles labelled with 111In twice, once with the shallow bolus technique (volumetic lung depth 76+/-20 mL ([+/- SD]) and once with the extremely slow inhalation technique (0.05 L/s). The radioactivity in the lungs was measured at 1 and 24 hours as well as at 1, 2, and 3 weeks after both inhalations. The 24-hour lung retention a percentage of lung deposition was significantly lower for the bolus inhalation, 46%+/-9% (+/- SD) than for the extremely slow inhalation, 56%+/-11%. The retention after 21 days as a percentage of the 24-hour retention was 55%+/-9% for the shallow bolus inhalation and 56%+/-10% for the extremely slow inhalation. Also within the subjects, clearance was similar for the 2 modes of inhalation. Deposition of particles inhaled with the 2 modes of inhalation was calculated with 2 model, one being based on Monte (Carlo particle transport together with an asymmetric lung model. Deposition predicted with this model agreed well with the experimental data under the assumption that there are large retained fractions only in small ciliated airways (bronchioli) and not in large ones. For the bolus inhalation, the model predicted 43% to 50% deposition in the bronchial (BB) region of initial lung deposition, 33% to 38% in the bronchiolar (bb) region, and 16% to 22% in the alveolar region. For the extremely slow inhalation, the model predicted 31% to 34% deposition in the BB region, 45% to 47% in the bb region, and 21% to 22% in the alveolar region. In addition, it predicted about the same ratio between bb and alveolar depositions for the 2 modes of inhalation. Thus, both the experimental and theoretical data indicate that the shallow bolus particles to a considerable extent reach both the bb and the alveolar regions and that they do that at about the same extent as the particles inhaled extremely slow. This conclusion is concerning the experimental data based on the assumption that there are no large retained fractions in the BB region. Another interpretation of the similar clearance for the two modes of inhalation is that there are large retained fractions in both the BB and the bb regions and that individual charactristics of clearance of these fractions are of importance rather than the site of deposition. PMID- 11400863 TI - Persistent versus transient map kinase (ERK) activation in the proliferation of lung epithelial type 2 cells. AB - Type 2 pneumocytes are progenitor cells of alveolor epithelium and important for reepithelialization following lung injury. This study examined the role of persistent versus transient mitogen-activated protein (MAP) kinase (extracellular signal-regulated kinase; ERK) in type 2 cell proliferation. Three different types of agents; epidermal growth factor (EGF), 12-O-tetradecanoylphorbol-13-acetate (TPA), and fetal bovine serum (FBS) induced different patterns of ERK activation. FBS induced a strong and persistent MAP kinase response, whereas the effect of EGF was transient with a strong activation at 5 minutes and only a slight stimulation at 4 hours. The TPA response was more prolonged than the EGF response, but not by far as strong and persistent as the FBS response. Activation by EGF and TPA and the early response induced by FBS were strongly reduced by the MEK inhibitor PD98059. The sustained FBS-induced ERK activation was inhibited by approximately 50%. The total number of cell, the percentage of cells in S and G2/M phase of the cell cycle and the incorporation of 3H-thymidine into DNA were strongly increased in response to FBS, whereas EGF and TPA were without effect. The proliferation was reduced by approximately 50% after pretreatment with PD98059. The results indicate that a persistent ERK activation of a critical size leads to type 2 cell proliferation, and that the proliferative response may also depend on a MEK-independent ERK activation. PMID- 11400864 TI - A cell-based drug delivery system for lung targeting: I. Preparation and pharmacokinetics. AB - A drug-loaded tumor cell (DLTC) system has been developed for lung metastasis targeting drug delivery. Doxorubicin was loaded into B16-F10 murine melanoma cells (96 microg/10(6) cells). The loading process led to the death of all the carrier cells. The diameter of DLTC was 15.03+/-2.36 microm (mean +/- SD). The amount and rate of doxorubicin being released from the DLTC mainly depended on the drug loading and carrier cell concentration. Over a 6-month storage in phosphate buffered saline (PBS) at 4 degrees C, the decrease in intracellular drug concentration and the carrier cell number were less than 25% and 5%, respectively. After a bolus injection of 30 microg doxorubicin in either DLTC form or free solution into the mice tail veins, drug deposit in the lung from DLTC was 3.6-fold of that achieved by free drug solution. The latter resulted in higher drug content in liver and spleen. Extensive trypsinization of DLTC reduced its lung targeting effect by 30%, and the density of surface adhesion molecule GM3 on DLTC surface by 25%. In conclusion, this DLTC system demonstrated a lung targeting activity that may be partially attributed to its specific surface characteristics. PMID- 11400865 TI - A cell-based drug delivery system for lung targeting: II. Therapeutic activities on B16-F10 melanoma in mouse lungs. AB - The in vitro and in vivo anticancer activities of doxorubicin-loaded B16-F10 murine melanoma cells (DLTC) were evaluated. DLTC showed similar growth inhibitory effects against live B16-F10 cells with doxorubicin solution in cell culture system, with the IC50 of 0.11 microM and 0.17 microM, respectively. However, DLTC demonstrated higher effectiveness than the free solution in treating mouse lung cancer caused by live B16-F10 cells. Syngeneic C57BL mice were inoculated intravenously with live B16-F10 cells first, and then received daily treatment of intravenous injections of doxorubicin in either DLTC or free solution form. Compared with the control group treated with phosphate-buffered saline, DLTC eradicated almost all the lung cancer colonies (>99%), while the free solution form reduced the colonies by 61%, when the treatment was given at an early stage. If the treatment started after the establishment of micrometastatic colonies in the mouse lungs, DLTC and free solution treatment resulted in 85% and 30% cancer reduction, respectively. Additional experiments demonstrated that the reduction of lung cancer colonies by DLTC was related to the initial treatment time: the earlier the treatment, the greater the effect. In conclusion, DLTC showed better therapeutic outcomes than free solution form in treating lung cancer of our animal model. PMID- 11400866 TI - Development of poly(Lactic acid)/chitosan co-matrix microspheres: controlled release of taxol-heparin for preventing restenosis. AB - Smooth muscle cell proliferation plays a major role in the genesis of restenosis after angioplasty or vascular injury. Controlled release of appropriate drugs alone and in combinations is one approach for treating coronary obstructions, balloon angioplasty, restenosis associated with thrombosis, and calcification. We demonstrated the possibility of encapsulating taxol-loaded polylactic acid (PLA) microspheres within heparin-chitosan spheres to develop a prolonged release co matrix form. The in vitro release profile of taxol and heparin from this co matrix system was monitored in phosphate buffered saline pH 7.4, using an ultraviolet spectrophotometer. The amount of taxol/heparin release was initially much higher, followed by a constant slow release profile for a prolonged period. The initial burst release of taxol (15.8%) and heparin (32.7%) from the co-matrix was modified with polyethylene glycol coatings (13.5% and 25.4%, respectively, for 24 hr). From scanning electron microscopy studies, it appears that these drugs diffuse out slowly to the dissolution medium through the micropores of the co-matrix. However, the surface micropores were modified with polyethylene glycol (PEG) coatings for a constant slow release profile. This PEG-coated PLA/chitosan co-matrix may target drug combinations having synergestic effects for prolonged periods to treat restenosis. PMID- 11400867 TI - Probing the dynamics of matrix hydration in the presence of electrolytes. AB - The aim of our work was to probe the mechanisms associated with induced matrix stiffening via textural analysis as a consequence of in situ electrolyte interactions within hydroxypropyl-methylcellulose (HPMC) and polyethylene oxide (PEO) matrices in relation to their role in controlling the release of highly soluble drugs such as diltiazem hydrochloride (>50% water soluble at 25 degrees C). The dynamics of HPMC and PEO matrix swelling during hydration in the presence of appropriate electrolytes intended to induce constant drug release rates from simple monolithic systems are influenced by continuously shifting peripheral matrix stiffening toward the matrix core in a manner dependent on electrolyte content and hydration time. Matrix erosion for HPMC and PEO controls (i.e., without electrolyte) follow linear dissolution kinetics (r2 > 0.97), while formulations with electrolyte characteristically undergo a square root of time decline in weight. The swelling potential of the electrolyte-containing matrices, influenced by the boundary infiltration process, reflected considerable suppression during the first 2 hr of exposure to medium, while subsequent events differed in both polymers. In view of these differences, simultaneous measurements in textural transitions and electrolyte conductivity showed that PEO has a higher affinity for water molecules than does HPMC. PMID- 11400868 TI - Controlled release tacrine delivery system for the treatment of Alzheimer's disease. AB - Alzheimer's disease is a neurodegenerative condition that affects approximately 5 million people and is the fourth leading cause of death in America. Tacrine is one of the three drugs approved by the FDA for the treatment of Alzheimer's disease. However, the drug has a short biologic half-life of 2-3 hr and gastrointestinal, cholinergic, and hepatic adverse reactions that are associated with high doses of the drug. The aim of our study was to formulate a controlled release delivery system of tacrine that could be used to minimize the side effects associated with the drug. Microparticles of tacrine were formulated using poly(D,L-lactide-co-glycolide) (PLG). PLG and tacrine were dissolved in mixed organic solvents and added to a polyvinyl alcohol solution that was stirred at a constant rate. The organic solvent was evaporated overnight and the formed microparticles were collected by filtration, dried, and sieve-sized. The effects of such formulation variables, as molecular weight of polymer, stir speed during preparation, and drug loading on encapsulation efficiency (EEF), and in vitro release profiles of tacrine were investigated. An increase in the molecular weight of polymer from 8,000 to 59,000 and 155,000 resulted in approximately 10 fold increase in EEF, but the rate of release decreased with increasing molecular weight. Stir speed during preparation had an effect on the EEF but not on the rate of release. Drug loading did not have a significant effect on the EEF but had an effect on the rate of tacrine release. The results suggest that tacrine could be delivered at controlled levels for weeks for the treatment of Alzheimer's disease. PMID- 11400869 TI - Formulation and characterization of immunoreactive tetanus toxoid biodegradable polymer particles. AB - Poly lactide-co-glycolide and polylactide polymer particles entrapping immunoreactive tetanus toxoid (TT) were prepared with a view to developing a single shot controlled release vaccine formulation. Denaturation of TT by dichloromethane (DCM) during primary emulsification stage of particle formulation was minimized by incorporation of an optimal amount of rat serum albumin (RSA) in the internal aqueous phase. Incorporation of RSA as a stabilizer during the primary emulsification stage of polymer particle formulation protected the immunoreactivity of TT, enhanced its encapsulation efficiency and also led to uniform polymer particle formation. Use of sonication, both during primary and secondary emulsification processes, resulted in formation of nanoparticles whereas microparticles were formed when the secondary emulsion was carried out by homogenization. Immunoreactive TT particles made from different polymers incorporating stabilizers released antigen continuously for more than four months in vitro. Single injection of both type of particles encapsulating stabilized TT elicited anti-TT antibody titers in rats for more than five months, which was higher than that obtained with TT injected in saline. Anti-TT antibody titers in vivo were in accordance with the in vitro release characteristics of immunoreactive TT from the particles. Immune responses with hydrophobic polymer particles were better than those made using hydrophilic polymers. These results indicate the importance of protecting the immunoreactivity of TT during formation of polymer particles for sustained and improved antibody response. PMID- 11400870 TI - High-energy extracorporeal shock wave treatment of nonunions. AB - Forty-three consecutive patients who did not have healing of tibial or femoral diaphyseal and metaphyseal fractures and osteotomies for at least 9 months after injury or surgery were examined prospectively for use of high-energy extracorporeal shock waves. Former treatment modalities (cast, external fixator, plate osteosynthesis, limitation of weightbearing) remained unchanged. In all cases a 99mTechnetium dicarboxyphosphonate regional two-phase bone scintigraphy was performed before one treatment with 3,000 impulses of an energy flux density of 0.6 mJ/mm2. Radiologic and clinical followups were done at 4-week intervals starting 8 weeks after shock wave treatment. The success criterion was bridging of all four cortices in the anteroposterior and lateral radiographic views, in oblique views, or by conventional tomography. An independent observer described bony consolidation in 31 of 43 cases (72%) after an average of 4 months (range, 2 7 months). Twenty-nine of 35 (82.9%) patients with a positive bone scan had healing of the pseudarthrosis compared with two of eight (25%) patients with a negative bone scan. Six of these eight patients with negative scans were heavy smokers. No complications were observed. High-energy shock wave therapy seemed to be an effective noninvasive tool for stimulation of bone healing in properly selected patients with a diaphyseal or metaphyseal nonunion of the femur or tibia. Additional controlled studies are mandatory. PMID- 11400871 TI - Effect of shock wave therapy on acute fractures of the tibia: a study in a dog model. AB - The effect of shock wave therapy on acute fractures of the tibia was studied in eight adult dogs. A fracture with a 3-mm gap was created in both tibias and the fractures were fixed internally with a small metallic plate and screws. Each of the right limbs received 2,000 impulses of shock waves at 14 kV whereas the left limbs were used as controls. The evaluations included the callus formations based on serial radiographic examinations at 1, 4, 8, and 12 weeks and histologic examinations at 12 weeks for tissue distribution including bone tissues. Based on radiographic findings, there was no statistically significant difference in the amount of callus formation between the treated and the control groups at 8 weeks or less. However, the radiographic findings at 12 weeks statistically showed more callus formations in the treated group. In histologic examinations, there was significantly more cortical bone formation in the treated group at 12 weeks and the bone tissues were thicker, denser, and heavier. Shock wave therapy enhanced callus formation and induced cortical bone formation in acute fractures in dogs at 12 weeks. The effect of shock wave therapy seemed to be time-dependent at 3 months. PMID- 11400872 TI - High-energy shock wave treatment of femoral head necrosis in adults. AB - Adults with Stages I to III osteonecrosis of the femoral head present an overall therapeutic challenge. The objective of the current prospective study was to show the effectiveness of high-energy shock wave therapy in treatment of patients with Association Research Circulation Osseous Stage I to Stage III necrosis of the femoral head by assessing clinical and magnetic resonance imaging results. The current study presents the results of 22 patients with femoral head necrosis 1 year after shock wave treatment. The study population consisted of 10 women and 12 men with an average age of 54.9 years (+/- 12.3). The scores achieved on the visual pain analog scale decreased from 8.5 before treatment to 1.2 after 1 year. Simultaneously, the Harris hip score increased from 43.3 to 92 points. Magnetic resonance imaging visualization of a distinct zone of sclerosis around the necrotic area remaining after treatment with extracorporeal shock wave therapy indicated therapeutic failure. The results obtained so far with high-energy shock wave therapy in these patients suggest that this method may offer an alternative to invasive treatment modalities for femoral head necrosis. A noninvasive and moderately priced method then would be available for the treatment of femoral head necrosis in the early stages of the disease process. PMID- 11400873 TI - Elastofibroma in shoulder osteoarthritis: a theoretical concept of the etiology. AB - A case of unilateral, subscapular elastofibroma dorsi secondary to degenerative osteoarthritis in the ipsilateral glenohumeral joint is presented. A 69-year-old woman had experienced symptoms of osteoarthritis in the right shoulder since contracting septic arthritis when she was 7 years old. The patient noticed a soft tissue mass in the right subscapular region when she was 65 years old. The range of motion of the glenohumeral joint was severely restricted. Histopathologic examination of the excised mass revealed elastofibroma. The authors think the excessive scapulothoracic motion was important in formation of the lesion. This case indicates that elastofibroma is not a true neoplasm but a reactive lesion formed by repetitive minor trauma. PMID- 11400874 TI - Anterior intramuscular transposition with ulnar nerve decompression at the elbow. AB - The effectiveness of decompression and anterior intramuscular transposition of the ulnar nerve for treatment of severe cubital tunnel syndrome was evaluated. A consecutive series of 39 anterior intramuscular transpositions were reviewed. One surgeon performed the transpositions between 1993 and 1997 in 34 patients who presented with clinically severe cubital tunnel syndrome. Clinical outcome and satisfaction with surgery were assessed. The results showed early clinical improvement of 77% of patients (mean followup, 3.34 months). With repeated assessments later, the same group of patients had clinical improvement of 62% (mean followup, 30.9 months). Patients younger than 50 years, individuals who underwent external neurolysis, or patients who had a previously failed subcutaneous transposition had fewer satisfactory results. PMID- 11400875 TI - Minimal access surgery in managing anterior lumbar disorders. AB - Traditional anterior lumbar surgery usually requires a long and sometimes painful skin incision. The current study evaluated the feasibility and safety of minimal access surgery for anterior lumbar disorders, emphasizing indications, operative technique, and the minimum 2-year followup results. From May 1996 to December 1997, the authors used this technique on 25 patients whose indications for surgery included syndromes of failed back surgery, selected cases of lumbar disc herniations, tuberculous or pyogenic spondylitis, selected spondylolisthesis, and vertebral tumors. In 23 of 25 patients, the site of interest was approached through a left flank incision, regardless of the laterality of the lesion. The mean length of the main incision was 5 cm. There were no injuries to great vessels or any neurologic deterioration after the procedures. Solid interbody fusion could be identified radiographically between 3 and 6 months after surgery. At a mean followup of 39.6 months, nine patients had excellent clinical outcomes, 11 patients had good outcomes, two patients had fair outcomes, and one patient had a poor outcome. The authors think such minimal access surgery is simple, effective, and safe for anterior lumbar disorders. The merits of the current technique include no need for endoscopic, microscopic, or complex surgical instruments, a lower amount of radiation exposure during surgery, and a shortened learning curve because the approach is similar to the anterior open lumbar technique, although the skin incision is only 5 cm in length. PMID- 11400876 TI - Supportive strut grafts for diaphyseal bone defects in revision hip arthroplasty. AB - Large femoral cortical strut allografts were used to provide structural support of the femur in 20 patients in whom uncemented, extensively porous-coated, press fit revision femoral stems were applied. The mean followup was 4.7 years. Radiographic analysis of the patients who had revision surgery revealed that in all 20 patients, the cortical femoral strut allografts showed incorporation. Small areas of graft resorption were observed in only two patients. Seventeen of the uncemented femoral revision stems radiographically showed bone ingrowth. Three of the revision femoral stems were interpreted as being fixed only by fibrous ingrowth after early subsidence after surgery. There were no reoperations. Before surgery, all patients were unable to walk and had severe pain. At followup, all patients were ambulatory and had considerable improvement in pain relief and in their ability to walk. The mean Harris hip score after surgery was 75 (range, 57.5-92). Complications consisted of one peroneal nerve palsy, which resolved; one deep venous thrombosis; one gastrointestinal ulcer; and one case of pneumonia. Supportive cortical strut allografts represent an alternative to the use of circumferential bulk allografts and total femoral replacements in patients with large combined proximal and diaphyseal bone defects. PMID- 11400877 TI - Middle-term results of Salter innominate osteotomy. AB - This study evaluated middle-term clinical and radiographic results of Salter innominate osteotomy for treatment of dysplastic, subluxated, or congenitally dislocated hips. Thirty-five hips in 33 patients with a mean age at index surgery of 4 years 7 months were followed up for an average of 16.5 years. Radiographic center-edge angle of Wiberg, acetabular index, and regularity of the femoral head were evaluated. Fifteen (43%) hips were in Group I, 11 (31%) hips were in Group II, six (17%) hips were in Group III, and three (9%) hips were in Group IV according to the Severin classification. Significant differences were found in the preoperative and recent center-edge angle (average, 1.1 degrees and 28.8 degrees in the excellent and good groups versus -5.3 degrees and 15.0 degrees in the fair and poor groups) and regularity of the femoral head (very irregular versus others). The current results indicate that after middle-term followup, Salter innominate osteotomy is an effective treatment. The authors conclude this procedure is indicated in patients who meet the criteria proposed by Salter. PMID- 11400878 TI - Correlation of estradiol in pregnancy and anterior cruciate ligament laxity. AB - The purpose of this study was to determine whether a correlation existed between estradiol in pregnancy and laxity of the anterior cruciate ligament by measuring anterior tibial translation. All patients underwent measurement of anterior tibial translation using KT-1000 knee arthrometer testing and serum estradiol determination during the third trimester of pregnancy and postpartum. Forty knees were studied. The average serum estradiol levels decreased from 10,755.0 ng/L to 50.3 ng/L. There was an average decrease anterior tibial translation with a manual maximum displacement of 3.0 mm (range, 1 mm-5 mm) from the first to second examinations. Average measurement of anterior tibial translation in pregnant women showed a statistically significant increase in laxity in the third trimester of pregnancy compared with the postpartum laxity. The results of this study show that high serum estradiol levels during the third trimester of pregnancy correlate with increased anterior tibial translation and that this anterior tibial translation decreases with the return of serum estradiol to nonpregnant levels. PMID- 11400879 TI - Radial displacement of the medial meniscus in varus osteoarthritis of the knee. AB - The macroscopic and histologic findings for 31 medial menisci and medial tibial plateaus obtained during total knee arthroplasty were examined to clarify the etiology and progression of varus osteoarthritis. Medial menisci were preserved fairly well in cases of severe osteoarthritis in which the medial joint space had already disappeared. The anterior segment was preserved in 26 (84%) menisci and the posterior segment was preserved in 11 (35%). The medial meniscus may have been preserved because of its radial displacement. Exposure of subchondral bone of the medial tibial plateau occurred in all 31 knees. The exposure of subchondral bone was centered in the anterior, middle, and posterior in nine, 10, and 12 medial tibial plateaus, respectively. There was a mechanical inconsistency between the pattern of preservation of the medial menisci and the location of exposure of subchondral bone on the medial tibial plateaus. The inconsistency reflects that the segment of the medial meniscus on which the excessive load was considered to exist was preserved fairly well. The authors' hypothesis for explaining this inconsistency is that radial displacement of the medial meniscus precedes narrowing of the medial joint space during progression of varus osteoarthritis, so that the displaced meniscus is saved from severe degeneration or attrition. PMID- 11400880 TI - Diagnosing deltoid injury in ankle fractures: the gravity stress view. AB - In the lateral malleolar ankle fracture without talar shift there is much uncertainty regarding the diagnosis of deltoid ligament injury severe enough to require surgical treatment. The current study evaluated the mechanical integrity of the ankle using a novel gravity-stress mortise radiographic view, which is practical for clinical use. Eight cadaveric lower extremities were tested under the following conditions: (1) intact ankle, (2) distal fibular oblique osteotomy, (3) plated fibula after osteotomy, (4) transection of the superficial deltoid with fibula osteotomized or plated, and (5) all possible combinations of deep deltoid transection with superficial deltoid transected or repaired and fibula osteotomized or plated. For each condition, a mortise radiograph was taken of the specimen while it was mounted horizontally, lateral side down. Fibular osteotomy with or without transection of the superficial deltoid did not alter the mortise radiograph appearance of the ankles. With combined deep and superficial deltoid transection and fibular osteotomy, the talus always (eight of eight specimens) showed a lateral shift of 2 mm or greater and a valgus tilt of 15 degrees or more. The gravity stress view of the ankle was found to reproducibly document destabilizing deltoid ligament damage. PMID- 11400881 TI - Application of shock waves in medicine. AB - Extracorporeal-generated shock waves were introduced approximately 20 years ago to disintegrate kidney stones. This treatment method substantially changed the treatment of urolithiasis. Shock waves have become the treatment of choice for kidney and ureteral stones. Urology, however, is not the only medical field for the potential use of shock waves for problems. Shock waves subsequently have been used in orthopaedics and traumatology to treat various insertional tendinopathies (enthesiopathies) and delayed unions and nonunions of fracture. Shock wave application also has been used in the treatment of tendinopathies in veterinary conditions (race horses). The concept of orthopaedic disorders is that shock waves stimulate or reactivate healing processes in tendons, surrounding tissue and bones, probably through microdisruption of avascular or minimally vascular tissues to encourage revascularization, release of local growth factors, and the recruitment of appropriate stem cells conducive to more normal tissue healing. The current author will give an overview of history and basic research of the application of shock waves in medicine. PMID- 11400882 TI - Posteroplantar release for congenital clubfoot in children younger than 1 year. AB - One hundred thirty-three resistant congenital clubfeet in 93 patients between 3 and 10 months of age were operated on using a standardized posteroplantar release. Clinical and radiographic assessments were done with a mean followup of 7 years 4 months (range, 3-12 years). Using the McKay score, 79.7% of the surgically treated clubfeet were classified as having a good or excellent result. Three patients had relapse of their clubfoot that required additional surgery. Seventeen feet in 15 patients had residual forefoot adduction at the time of followup. The radiographs showed that the early posteroplantar release led to sufficient hindfoot correction in all but the three patients who had relapse of the clubfoot. With this standardized surgical treatment, satisfactory results can be achieved in most patients younger than 1 year with congenital clubfoot. However, in patients with persistent talonavicular subluxation after conservative treatment, an additional talonavicular release combined with the posteroplantar release is recommended. PMID- 11400883 TI - Is radiotherapy important for low-grade soft tissue sarcoma of the extremity? AB - Radiotherapy and limb-preserving surgery has replaced amputation and compartmental resection for treatment of soft tissue sarcomas. However, the role of radiotherapy in low-grade tumors remains unclear. This study reviews the outcomes of 132 patients who received multimodality treatment for low-grade soft tissue sarcoma. Large primary tumors (> 5 cm) and the absence of radiotherapy correlated with local recurrence. Radiotherapy was most effective in patients operated on with marginal margins. Patients who were treated with wide surgical margins or had small tumors (< or =5 cm) showed no benefit with adjuvant radiotherapy. Size greater than 5 cm and local recurrence correlated with metastasis. Radiotherapy appears to be important in the management of low-grade soft tissue sarcoma. The principles of local treatment for low-grade soft tissue sarcoma should be the same as for high-grade tumors with a combination of surgery and adjuvant radiotherapy. In a subset of patients with small and widely excised tumors, consideration may be given to withholding radiotherapy. Local recurrence and metastasis from low-grade soft tissue sarcoma may occur as long as 1 decade after primary tumor resection. Long-term review of patients with low-grade tumors may be indicated. PMID- 11400884 TI - Extracorporeal shock wave therapy in orthopaedics. PMID- 11400885 TI - Osteoid osteoma that occurred in a former fracture site. AB - Diagnosis of osteoid osteoma often is delayed, despite its high incidence, because of similarities in presenting symptoms with other pathologic entities. The current case report describes a posttraumatic osteoid osteoma. Three years after osteosynthesis of a distal tibial fracture an osteoid osteoma was diagnosed at the former fracture site. After excluding osteomyelitis as a possible diagnosis, the tumor was excised successfully. Based on current knowledge of the pathogenesis of osteoid osteoma, it is unlikely that the lesion observed in the patient was attributable to the previous fracture. PMID- 11400886 TI - Limb salvage surgery with vascular reconstruction. AB - Vascular reconstruction and limb salvage surgery has been the authors' preferred approach when malignancy involves major vessels of the extremities. Treatment of 16 patients involved resection, with vascular grafting in 14 patients and rotationplasty in two patients. The major vessels were surrounded by tumor in six patients, nearly encased in three patients, invaded by tumor in four patients, and widely contaminated by intralesional surgery in three patients. The tumor stage included one Stage IB, 12 Stage IIB, two Stage IIIB sarcomas, and one multiply recurrent carcinoma. The largest average tumor dimension was 9.5 cm, and the length of grafting was 14 cm. Major nerves were sacrificed in eight (50%) patients, flaps or muscle transfers were done in seven (44%), chemotherapy was administered in nine (56%), radiation therapy was used in four (25%), and pulmonary metastasectomy was done in two (12%). At a mean followup of 56 months, 50% (eight of 16) of patients were alive without disease. Local recurrence was 12% (two of 16 patients), and infection was 12% (two of 16 patients). Limb salvage was achieved in 88% (14 of 16 patients), and functional status was judged good or excellent in 81% (13 of 16 patients). The complication rate observed in this subset of patients is significant, yet local control and the incidence of major complications was acceptable. Results observed from this series and data gathered from the literature clearly indicate that patients can avoid amputation, despite malignant involvement of major vessels to their extremities. PMID- 11400887 TI - Computerized analysis of radiographs in Colles' fractures. AB - Several radiographic parameters correlate with clinical outcome in distal radius fractures. This study describes a new, computerized routine that automatically defines the longitudinal axis of the radius, deriving the radial tilt, radial width, and radial height on anteroposterior radiographs of the wrist. The intraobserver and interobserver variability is reported in a series of 33 anteroposterior radiographs of the wrist. An edge-finding filter is used to determine accurately the cortices in several lines of each image. The midpoints between these two edges are used to determine the best-fit line, thereby defining the longitudinal axis of the bone. The operator marks the tip of the radial styloid and the lunate angle of the radius at the distal radioulnar joint. Radial shortening, widening, and loss of angle are calculated. The 33 radiographs were measured twice by two independent observers, and 12 sets of paired readings were analyzed using the two-tailed paired t test. The mean difference between readings was reduced to a fraction of degree or millimeter, virtually eliminating the intraobserver and interobserver errors. Thus, the current study shows the computer is a reliable and effective instrument for measuring radiographs. The computer-aided system of measurements can be extended to prostheses, fractures, and to other orthopaedic measurements. PMID- 11400888 TI - Shock wave therapy (Orthotripsy) in musculoskeletal disorders. AB - Extracorporeal shock wave therapy, which now is used routinely for urolithiasis, has gained increasing acceptance in Europe for some musculoskeletal problems and has led to the inception of clinical studies in the United States. The authors have reviewed the available literature to assess the biologic effects of shock waves on human musculoskeletal tissues, the credibility of published studies on therapeutic applications, and the potential for more widespread application of this modality to various skeletal and near-skeletal disorders. The primary advantage of extracorporeal shock wave therapy is its noninvasive nature and seemingly minimal complications when applied to musculoskeletal tissues. PMID- 11400889 TI - Contamination risk of the surgical team through ROBODOC's high-speed cutter. AB - During cutting of the femoral cavity in the ROBODOC procedure, an aerosol cloud of irrigation fluid, blood, and tissue debris was seen. This cloud potentially is contaminated with bacterial and viral vectors, which is an infection risk for the surgical team. A flat and a ball cutter were tested in four standard situations macroscopically with a colored solution. In a second experiment, the cutter was exposed to a fluid contaminated with Staphylococcus aureus, and bacterial room contamination was detected using standard cultures. The aerosol cloud was seen in a 6 x 3.6-m area. Extension and concentration varied, depending on the irrigation situation. ROBODOC's high-speed cutter produces an aerosol cloud in an area in which all members of the surgical team are affected. Sufficient protection is necessary for everyone in the operating room. PMID- 11400890 TI - Histologic change of the meniscus and cartilage tissue after meniscal suture. AB - The influence of suturing on cell infiltration into the meniscus and surrounding tissue is not well known. Histologic changes in the meniscus after suturing and prediction of histologic changes using magnetic resonance imaging were studied. Forty knees in 20 mongrel dogs were studied using four types of 4-0 suture material: nonabsorbable monofilament, nonabsorbable braided, absorbable monofilament, and absorbable braided. Each type of suture material was used for meniscal suture on eight knees each. The other eight knees were not subjected to meniscal suture. Specimens obtained at 1 and 3 months were studied by magnetic resonance imaging and light microscopic analysis. The maximum width of change of meniscal tissue was measured. Changes from the nonabsorbable suture group were smaller than that of the absorbable suture group in magnetic resonance imaging. Histologic changes as seen by light microscope were larger in the absorbable group than in the nonabsorbable group at 1 and 3 months after surgery. Injury was found in the articular cartilage of the femoral condyle in the nonabsorbable braided suture group. Histologic changes were significantly greater when changes in magnetic resonance imaging signal intensity were larger. The current study showed that non-absorbable monofilament suture material results in the least damage to the meniscus and surrounding tissue. PMID- 11400891 TI - Bernese periacetabular osteotomy. PMID- 11400892 TI - The lithotriptor and its potential use in the revision of total hip arthroplasty. PMID- 11400893 TI - Treatment of painful heel syndrome with shock waves. AB - In a prospective clinical study, the effectiveness of shock waves on painful heel syndrome in 80 patients (20 men and 60 women) with an average age of 48 years was investigated. Six patients had bilateral treatments. Each treatment consisted of 1,000 impulses of shock waves at 14 kV. A 100-point scoring system (70 points for pain and 30 points for function) was used for evaluation. The intensity of pain was measured with a visual analog scale from 0 to 10. The overall results were no complaints in 20.6%, significantly better in 52.9%, slightly better in 17.6%, and unchanged in 8.8% of 64 patients (68 heels) with 12 weeks followup; no complaints in 59.3%, significantly better in 27.7 %, slightly better in 13% of 52 patients (54 heels) with 6 months followup. None of patients' symptoms became worse. Seventeen patients (18 heels) who did not respond favorably to the first treatment had significantly better results after a second treatment. There were no device-related problems, and no systemic or local complications. Shock wave treatment is a new modality of therapy that is safe and effective in the treatment of patients with painful hell syndrome. PMID- 11400894 TI - Shock wave therapy for chronic proximal plantar fasciitis. AB - Three hundred two patients with chronic heel pain caused by proximal plantar fasciitis were enrolled in a study to assess the treatment effects consequent to administration of electrohydraulicall-generated extracorporeal shock waves. Symptoms had been present from 6 months to 18 years. Each treated patient satisfied numerous inclusion and exclusion criteria before he or she was accepted into this study, which was approved by the Food and Drug Administration as a randomized, double-blind evaluation of the efficacy of shock wave therapy for this disorder. Overall, at the predetermined evaluation period 3 months after one treatment, 56% more of the treated patients had a successful result by all four of the evaluation criteria when compared with the patients treated with a placebo. This difference was significant and corroborated the fact that this difference in the results was specifically attributable to the shock wave treatment, rather than any natural improvement caused by the natural history of the condition. The current study showed that the directed application of electrohydraulic-generated shock waves to the insertion of the plantar fascia onto the calcaneus is a safe and effective nonsurgical method for treating chronic, recalcitrant heel pain syndrome that has been present for at least 6 months and has been refractory to other commonly used nonoperative therapies. This technology, when delivered using the OssaTron (High Medical Technology, Kreuz-lingen, Switzerland), has been approved by the Food and Drug Administration specifically for the treatment of chronic proximal plantar fasciitis. The results suggest that this therapeutic modality should be considered before any surgical options, and even may be preferable to cortisone injection, which has a recognized risk of rupture of the plantar fascia and recurrence of symptoms. PMID- 11400895 TI - Treatment of lateral epicondylitis of the elbow with shock waves. AB - In a prospective clinical study, the effectiveness of shock wave treatment for lateral epicondylitis in 56 elbows in 53 patients (27 men and 26 women) with an average age of 46 years was investigated. Three patients received treatment for both elbows. Each elbow was treated with 1,000 impulses of shock waves at 14 kV. A 100-point scoring system was used for evaluation including 40 points for pain, 30 points for function, 20 points for strength, and 10 points for elbow motion. The intensity of pain was measured using a visual analogue scale from 0 to 10. The overall results were 13.2% excellent, 44.7% good, 36.8% acceptable, and 5.3 unchanged in 35 patients with 12 weeks followup; 30.8% excellent, 42.3% good, and 26.9% acceptable in 25 patients with 24 weeks followup. Considerable improvement was observed from 6 weeks to 6 months after the treatment. None of the patients' symptoms became worse. The results of nine patients who also received a second treatment were good in three patients, acceptable in five patients, and unchanged in one patient. There was no device-related problems, systemic, or local complications. Shock wave therapy may offer a new and safer nonoperative treatment for patients with lateral epidoncylitis of the elbow. PMID- 11400896 TI - Correlations between the duration of pain and the success of shock wave therapy. AB - Patients who have had successful treatment for either chronic heel pain (plantar fasciitis) or humeral epicondylitis subsequently were evaluated for a comparable problem in the contralateral heel or elbow. Patients who had experienced symptoms in the contralateral heel or elbow for a shorter period were less likely to have a positive result from shock wave therapy than those patients who had received treatments for more chronic symptoms. PMID- 11400897 TI - Shock wave therapy versus conventional surgery in the treatment of calcifying tendinitis of the shoulder. AB - A prospective quasirandomized study was performed to compare the effects of surgical extirpation (Group I, 29 patients) with the outcome after high-energy extracorporeal shock wave therapy (Group II, 50 patients; 3,000 impulses of an energy flux density of 0.6 mJ/mm2) in patients with a chronic calcifying tendinitis in the supraspinatus tendon. Symptoms and demographic data of the two groups were comparable. According to the University of California Los Angeles Rating System, the mean score in Group I was 30 points with 75% good or excellent results after 12 months, and 32 points with 90% good or excellent results after 24 months. Radiologically, there was no calcific deposit in 85% of the patients after 1 year. In Group II, the mean score was 28 points with 60% good or excellent results after 12 months, and 29 points with 64% good or excellent results after 2 years. Radiologically, complete elimination of the deposit was observed in 47% of the patients after 1 year. Clinically, according to the University of California Los Angeles score, there was no significant difference between both groups at 1 year. At 2 years, there was a significantly better result in Group II. Both groups then were subdivided into patients who had a homogenous deposit as seen on radiographs and patients who had an inhomogenous deposit before treatment. Surgery was superior compared with high-energy shock wave therapy for patients with homogenous deposits. For patients with inhomogenous deposits, high-energy extracorporeal shock wave therapy was equivalent to surgery and should be given priority because of its noninvasiveness. PMID- 11400898 TI - Principles of shock wave therapy. AB - A shock wave is a transient pressure disturbance that propagates rapidly in three dimensional space. It is associated with a sudden rise from ambient pressure to its maximum pressure. A significant tissue effect is cavitation consequent to the negative phase of the wave propagation. The current authors summarize the basic physics of shock waves and the physical parameters involved in assessing the amount of energy delivered to the target tissue and in comparing the various high and low-energy devices being evaluated clinically for musculoskeletal applications. PMID- 11400899 TI - Treatment of calcifying tendinitis of the shoulder with shock wave therapy. AB - In a prospective clinical study, the effectiveness of shock wave treatment for calcifying tendinitis in 31 shoulders in 29 patients (14 women and 15 men) with an average age of 52 years was assessed. Each shoulder was treated with 1,000 impulses of shock waves at 14 kV. A 100-point Constant score system was used for evaluation. The overall results of 20 patients (21 shoulders) with 12 weeks followup were no complaints in 23.8%, significantly improved in 38.1%, some improvement in 14.3%, and unchanged in 23.8%. Considerable improvement was observed between 6 and 12 weeks. The results of seven patients with 24 weeks followup were no complaints in two patients, significantly improved in three, and unchanged in two patients. Radiographs showed complete elimination of the calcium deposits in six patients (28.6%), incomplete elimination in two patients (9.5%), and three patients (14.3%) had fragmentation of the calcium deposits. There was no recurrence of calcium deposits observed at 24 weeks. There was a correlation between the functional improvement and the elimination of calcium deposits. There were no device-related problems, systemic or local complications. Low-energy shock wave therapy may offer a new and safer additional nonoperative treatment for patients with calcifying tendinitis of the shoulder. PMID- 11400900 TI - Extracorporeal shock wave therapy of nonunion or delayed osseous union. AB - One hundred fifteen patients with nonunions or delayed fracture healing were treated with high-energy shock waves. After shock wave treatment, immobilization of the fracture also was done. The followup was at least 3 months and as long as 4 years. In 87 patients (75.7%), one treatment with shock waves resulted in bony consolidation with a simultaneous decrease in symptoms. Besides negligible local reactions (swelling, hematomas, petechial hemorrhages), no complications were observed. The treatment was noninvasive, and personnel and technical requirements were not problematic. The authors concluded that the application of extracorporeal shock wave therapy should be the first choice of treatment for patients with nonunions and delayed bone fracture healing. PMID- 11400901 TI - Treatment of nonunions of long bone fractures with shock waves. AB - A prospective clinical study investigated the effectiveness of shock waves in the treatment of 72 patients with 72 nonunions of long bone fractures (41 femurs, 19 tibias, seven humeri, one radius, three ulnas and one metatarsal). The doses of shock waves were 6,000 impulses at 28 kV for the femur and tibia, 3,000 impulses at 28 kV for the humerus, 2,000 impulses at 24 kV for the radius and ulna, and 1,000 impulses at 20 kV for the metatarsal. The results of treatment were assessed clinically, and fracture healing was assessed with plain radiographs and tomography. The rate of bony union was 40% at 3 months, 60.9% at 6 months, and 80% at 12 months followup. Shock wave treatment was most successful in hypertrophic nonunions and nonunions with a defect and was least effective in atrophic nonunions. There were no systemic complications or device-related problems. Local complications included petechiae and hematoma formation that resolved spontaneously. In the authors' experience, the results of shock wave treatment were similar to the results of surgical treatment for chronic nonunions with no surgical risks. Shock wave treatment is a safe and effective alternative method in the treatment of chronic nonunions of long bones. PMID- 11400902 TI - Rehabilitative management of post-stroke visuospatial inattention. AB - PURPOSE: Visuospatial inattention or sensory neglect is a common impairment following cerebrovascular accident and is thought to negatively impact on functional recovery and long-term outcome. This review examines various rehabilitative interventions available for the management of visuospatial inattention and the literature that supports their efficacy. METHOD: Medical literature review. RESULTS: Therapies geared toward improved visual scanning and 'retraining' of patients to attend to the neglected field are the mainstay of current therapy. While some studies support their efficacy, others have reported limited benefit. Newer approaches that may be efficacious include eye patching techniques, use of video feedback during therapy, training in visual imagery, and pharmacologic therapy with dopamine agonists. Protocols using vestibular, somatosensory and optokinetic stimulation have each been shown to produce transient improvements in neglect. However, therapeutic applications for these techniques have not been studied. CONCLUSIONS: A number of approaches toward the rehabilitation of post-stroke neglect are described in the literature. Further research is needed to better define which techniques may prove most beneficial. PMID- 11400903 TI - Positioning for stroke patients: a survey of physiotherapists' aims and practices. AB - PURPOSE: The survey was undertaken to identify current physiotherapy practice for positioning patients in the first week following stroke. METHOD: A postal questionnaire comprised of closed questions, was sent to 674 physiotherapists identified as working with patients in the first week following stroke, who were employed in 155 randomly selected NHS Trusts throughout England. RESULTS: Response rate was 73%. Specific positions were recommended during the first week following stroke by 98 % of respondents. The most common aims of positioning were modulation of muscle tone (93%), preventing damage to affected limbs (92%) and supporting and stabilizing body segments (91%). The positions sitting in an armchair, side lying on the non-hemiplegic side and side lying on the hemiplegic side were recommended by 98%, 96% and 92% of respondents respectively. The components of the positions considered as 'most important' varied between positions, proximal components were usually preferred to distal components. CONCLUSION: Positioning is still an important part of physiotherapy practice and therefore requires evaluation. The positions used and the aims of positioning identified by clinicians accord with those in the literature. However, there is a lack of consensus regarding key components of the positions. The positions identified in this study should now be systematically evaluated for their ability to achieve different aims. PMID- 11400904 TI - Early interdisciplinary rehabilitation programme for whiplash associated disorders. AB - PURPOSE: To assess the clinical outcomes of a multidisciplinary rehabilitation programme for early intervention of chronic whiplash associated disorders (WAD). The primary aim of the programme was to increase levels of activity and independence in patients suffering WAD. METHODS: Ninety subjects with purported chronic symptoms associated with whiplash associated disorder (WAD) were referred to a multi-centre multi-modal 5- and 8-week rehabilitation programme in 1997 1998, during which prospective and retrospective functional and psychological evaluations were conducted, the follow-up was to 6 months. RESULTS: Subjects indicated they were satisfied with the programme. Retrospective evaluation indicated increased ability to cope with and control pain and, to some extent, psychological aspects. The pain intensity in the neck and upper back were significantly decreased at 6 months follow-up. However, for most of the functional and psychological markers, no significant changes were found. CONCLUSIONS: A multi-modal rehabilitation programme for the chronic suffering attributed to WAD had positive effects according to several aspects of the retrospective evaluations, but according to most of the aspects evaluated prospectively the programme does not appear to have significant benefits. PMID- 11400905 TI - The impact of psychosocial 'markers' on the outcome of rehabilitation. AB - PURPOSE: The aim of this study was to identify substantial psychosocial events- 'psychosocial markers' (PMs) in the clients files and to correlate the PMs to the outcome of the rehabilitation, in order to find out whether this information had any impact on the outcome of vocational rehabilitation (VR). METHOD: Three hundred and seventy-two individuals were included, being an object of a VR measure during 1993-1994. All were impaired by either disc hernia back pain syndrome or other diseases of the loco-motor system. The PM was defined as any psychosocial loaded information of the person's life situation, not immediately related to the original set of medical problems. RESULTS: Twenty-seven PMs were identified among men and 36 for women. Alcohol related problems dominated for men and private life related problems for women. Only approximately 25% of the PM concerned work related matters. After 3 years the OR (odds ratio) for full disability pension was 2.6 (n.s.) for men with the PM and 4.3 (p < 0.05) for women. The outcome was independent of background factors such as unemployment, blue-collar work and low education. CONCLUSIONS: The PM was found to be an important and independent factor to impact the outcome of rehabilitation in women. PMID- 11400906 TI - An integrated physiotherapy/cognitive-behavioural approach to the analysis and treatment of chronic whiplash associated disorders, WAD. AB - PURPOSE: The aim of this paper was to describe a model for an integrated physiotherapy/cognitive-behavioural approach in the analysis and treatment of chronic WAD patients, as well as to evaluate the effectiveness of this approach in three experimental single case studies. METHOD: Three patients with a diagnosis of chronic WAD were included in the study. Psychological and physical functional analyses were used to describe the problem areas and as a basis for the management of WAD. A programme including learning of basic and applied skills, generalization, and maintenance was carried through. RESULTS AND CONCLUSION: The results showed that functional behavioural analyses can be useful in physiotherapy for structured patient assessment and in planning of treatment. It was also shown that physiotherapy integrated with cognitive behavioural components decreased the patients' pain intensity in problematic daily activities. PMID- 11400907 TI - Successful fitting of a prosthesis for a child amputee suffering from scleroderma. AB - A few anecdotal cases of limb amputations owing to scleroderma have been described in the literature. This article outlines the salient problematic features of a residual stump and provides the prosthetic considerations and appropriate design that eventually facilitated ambulation. PMID- 11400908 TI - Circadian rhythms of body temperature and motor activity in rodents their relationships with the light-dark cycle. AB - In rodents, the alternation of light and dark is the main synchronizer of circadian rhythms. The entrainment abilities of the LD cycle could be estimated by experimental modifications of the photoperiod and by following the subsequent temporal distribution of a circadian rhythm. The rate of reentrainment of a rhythm is determined by the nature of the studied variable, by the direction (advance or delay) and the magnitude (or value) of the phase shift. In rodents, core body temperature and motor activity are known to be well synchronized with each other under L:D 12:12 and under constant conditions (LL or DD). There are clear evidences that the circadian pattern of motor activity is generated by two oscillators, one from dusk signal and the other from dawn signal. Whether the circadian rhythms of body temperature and motor activity are generated by a common circadian mechanism or controlled by separate ones still remains unknown. The purpose of this review is to summarize the results obtained on the circadian rhythms of body temperature and motor activity throughout the daily cycle in order to clarify the relationships between these two functions. PMID- 11400909 TI - Brain fatty acids in perinatal asphyxia. AB - In hypoxic or ischemic states the release of fatty acids is proposed to have several harmful effects on brain structure and function. We therefore decided to study brain FFA in a simple, clinically related animal model resembling intrauterine perinatal asphyxia (PA). Cerebral blood flow (CBF), brain fatty acids (C14:0, C16:1, C16:0, C18:1, C1 8:0, sigma C), plasma glucose, lactate, beta-hydroxybutyrate (beta-OHB), non-esterified fatty acids (NEFA) and insulin were determined in PA and compared to the normoxic state. Brain C 14:0 FFA were not significantly different from normoxic rats. Brain FFA C 16:0 were comparable between groups but significantly decreased at 20 min of PA. C 18:0 FFA showed a trend to increase with the length of PA reaching significance at 10 min of asphyxia only and were declining at 20 min, however, not significantly. Brain C 16:1 and C 18:1 FFA concentrations were comparable between groups. The parameters cerebral blood flow, glucose and lactate showed a stepwise and significant increase with the length of PA, whereas beta-HOB, NEFA and insulin showed no changes. CBF, glucose and lactate showed a strong association whereas other parameters failed to correlate with each other. Only inconsistent trends of increased brain FFA were found and the association between brain glucose and brain FFA could be ruled out. Although CBF was manifold and significantly elevated in PA, brain FFA pattern suggests that the increase of CBF is obviously not mediated by brain FFA. We conclude that FFA may not be involved in the early phase-pathogenesis of PA. PMID- 11400910 TI - Protection from drug-induced hepatocellular changes by pretreatment with conjugating enzyme inhibitors in rats. AB - The present paper describes the role of conjugating enzymes in the development of hepatotoxicity after administration of repeated doses of a novel monoamine oxidase type-A (MAO-A) inhibitor, (5R)-3-[2-(( 1S)-3-cyano-1 hydroxypropyl)benzothiazol-6-yl]-5-methoxymethyl-2-oxazolidinone (E2011). The effects of pretreatment with three kinds of conjugating enzyme inhibitors on hepatic lesions induced by E2011 were evaluated in female Sprague-Dawley rats. The inhibitors used were 2,6-dichloro-4-nitrophenol (DCNP; inhibitor of sulfotransferase (ST)), pentachlorophenol (PCP; inhibitor of both ST and acetyltransferase (AT)) or ranitidine (inhibitor of UDP-glucuronosyltransferase (UDP-GT)). Two weeks treatment of E2011 alone at an oral dosage of 150 mg/kg induced hepatocellular changes characterized by nuclear enlargement. Daily pretreatment with DCNP (10 mg/kg, i.p.) enhanced the E2011-induced hepatocellular changes accompanied by single cell necrosis. On the other hand, the hepatotoxicity was clearly diminished by PCP (5 mg/kg, i.p.). Ranitidine pretreatment had no effect. Protection by PCP was attributed to the inhibitory effects of AT in addition to ST; it was considered that the hepatocellular changes caused by E2011 were largely dependent on the formation of acetyl conjugate(s). PMID- 11400911 TI - Increased tumor necrosis factor-alpha and nitric oxide production by rat macrophages following in vitro stimulation and intravenous administration of the delta-opioid agonist SNC 80. AB - Opioids alter immune function by binding to opioid receptors on cells of the immune system, or indirectly by acting on receptors within the central nervous system. Mu-selective opioid agonists are generally associated with immunosuppression, whereas delta-opioid receptor-selective agonists are commonly associated with immunopotentiation. We have previously shown that intracerebroventricular administration of the nonpeptide delta-opioid receptor agonist (+)-4-((alpha R)-alpha-((2S, 5R)-4-allyl-2, 5-dimethyl-1-piperazinyl)-3 methoxybenzyl)-N, N-diethyl-benzamide (SNC 80) did not alter certain parameters of immunocompetence. In the present study, we studied the in vitro and ex vivo effects of SNC 80 on rat macrophage and lymphocyte functions. We showed that SNC 80 at concentrations of 10(-7) M and 10(-6) M, significantly (P < 0.01) stimulated the in vitro production of tumor necrosis factor-alpha (TNF-alpha) (60 100% increase) and nitric oxide (34-67% increase) by resident and LPS-stimulated peritoneal macrophages. Similarly, intravenous administration of SNC 80 (6.8 mg/kg) significantly (P < 0.01) increased the production of TNF-alpha and nitric oxide (2- and 1.5-fold increases respectively, compared with saline-injected control) by LPS-stimulated splenic macrophages. In addition, intravenous injection of SNC 80 plus Con A potentiated ex vivo LPS-stimulated macrophage functions. SNC 80 could potentially be utilized in various clinical situations where immunosuppression is undesirable. PMID- 11400912 TI - Advanced glycation alters expression of the 67kDa laminin receptor in retinal microvascular endothelial cells. AB - The 67kDa laminin receptor (67LR) plays an important role in vascular cell function and dysfunction. The present study has examined 67LR expression in retinal microvascular endothelial cells after exposure to AGEs. Retinal microvascular endothelial cells were exposed to either AGE-BSA, or were grown on methylglyoxal-modified laminin or Matrigel and expression of 67LR analysed by Western Blotting and RT-PCR/Southern blotting. Western blotting of plasma membrane and RT-PCR/Southern blotting revealed a significant upregulation of 67LR protein/mRNA expression after exposure to AGEs (p<0.05-0.01). The results show that 67LR is upregulated in cells exposed to AGEs and suggests a previously unrecognised role for this receptor in retinal microvascular endothelial cell interaction with glycated substrates. PMID- 11400913 TI - Diurnal variation of leptin entry from blood to brain involving partial saturation of the transport system. AB - The blood-brain barrier (BBB) regulates the amount of peripherally produced leptin reaching the brain. Knowing that the blood concentration of leptin has a circadian rhythm, we investigated whether the influx of leptin at the BBB followed the same pattern in three main sets of experiments. (a): The entry of 125I-leptin from blood to brain was measured in mice every 4 h, as indicated by the influx rate of 125I-leptin 1-10 min after an iv bolus injection. The blood concentration of endogenous leptin was measured at the same times. Blood leptin concentrations were higher at night and early morning (peak at 0800 h) and lower during the day (nadir at 1600 h). By contrast, the influx of 125I-leptin was fastest at 2000 h and slowest at 0400 h. Addition of unlabeled leptin (1 microg/mouse) significantly decreased the influx rate of 125I-leptin at all time points, indicating saturability of the transport system. The unlabeled leptin also abolished the diurnal variation of the influx of 125I-leptin. (b): The entry of 125I-leptin into spinal cord was faster than that into brain and showed a different diurnal pattern. The greatest influx occurred at 2400 h and the slowest at 0800 h. In spinal cord, unlike brain, unlabeled leptin (1 microg/mouse) neither inhibited the influx of 125I-leptin nor abolished the diurnal rhythm. (c): Higher concentrations of unlabeled leptin (5 microg/mouse) inhibited the uptake of 125I-leptin in spinal cord as well as in brain, but not in muscle. This experiment measured uptake 10 min after iv injection at 0600 h (beginning of the light cycle) and 1800 h (beginning of the dark cycle). Thus, influx of 125I leptin into the CNS shows diurnal variation, indicating a circadian rhythm in the transport system at the BBB, saturation of the leptin transport system shows differences between the brain and spinal cord, and blood concentrations of leptin suggest that partial saturation of the transport system occurs at physiological concentrations of circulating leptin, contributing to the differing diurnal patterns in brain and spinal cord. Together, the results show that the BBB is actively involved in the neuroendocrine regulation of feeding behavior. PMID- 11400914 TI - Endothelin-1, endothelin-1 receptors and cardiac natriuretic peptides in failing human heart. AB - Endothelin (ET)-1 is a potent vasoconstrictor peptide produced in the myocardium that can exert important effects on cardiac myocyte growth and phenotype; cardiac natriuretic peptides (ANP and BNP) are known to act as physiological antagonists of ET-1. In this study a comparative determination of ET-1 receptors and of the local productions of ET-1 and of ANP and BNP was made in different sites of failing and nonfailing hearts. Tissue from right and left atrium, right and left ventricle and interventricular septum from seven adult heart transplant recipients with end-stage idiopathic dilated cardiomyopathy (functional class III and IV, with ejection fraction < 35%) and from four postmortem subjects without cardiac complications was analyzed. In failing hearts we observed a tendency to increase of density of binding sites, most evident in left ventricle (62.6+/-22.6 fmol/mg protein vs. 29.0+/-3.3, mean +/- SEM, p = ns). A prevalence of ET-A subclass, observed in all samples, resulted more pronounced in failing hearts where this increase, found in all the cardiac regions, was more evident in left ventricle (p = 0.0007 vs nonfailing hearts). The local concentrations of ET-1, ANP and BNP resulted significantly increased in failing hearts with respect to controls in all sides of the heart. In failing hearts we have observed a tendency to increase in endothelin receptor density mainly due to a significant upregulation of ET-A subtype and a parallel increase of the tissue levels of ANP, BNP and ET-1 indicating an activation of these systems in heart failure. PMID- 11400915 TI - Discrimination between peptide and non-peptide opioid agonists on the transcription of opioid receptors in two cell lines. AB - The aim of the present study was to characterize the effects of prolonged use of peptide mu- and delta-receptor agonists [D-Ala2, N-me-phe, Gly5-ol]-enkephalin (DAMGO) and [D-Pen2, D-Pen5]-enkephalin (DPDPE) and non-peptide agonists ohmefentanyl (OMF) and BW373U86 on the transcription of opioid receptors of cultured NG108-15 cell and SHSY5Y cells, respectively using the method of reverse transcription-polymerase chain reaction (RT-PCR). It was found that (1) The abundance of mu- and delta- receptor mRNA decreased significantly up to 48h after the administration of DAMGO and DPDPE, respectively; whereas the inhibitory effect of OMF and BW373U86 lasted only for 24h; (2) DAMGO and DPDPE produced a significant decrease of the mRNA coding for mu-receptor and delta-receptor at concentrations as low as 10(-8) mol/L and 10(-6) mol/L, respectively, whereas OMF and BW373U86 were effective at concentrations one order of magnitude higher, respectively. These results suggested that (1) Long-term administration of either peptide or non-peptide opioid agonist to cultured cell line produced a significant decrease of the gene expression of opioid receptor at transcription level. (2) The effect of peptide agonists was stronger and lasted longer than that of corresponding nonpeptide agonists. PMID- 11400916 TI - The reduction of NADH ubiquinone oxidoreductase 24- and 75-kDa subunits in brains of patients with Down syndrome and Alzheimer's disease. AB - NADH: ubiquinone oxidoreductase (complex I), one of the most complicated multi protein enzyme complexes, is important for energy metabolism because it is the initial enzyme of the mitochondrial respiratory chain. Deficiency of complex I is frequently found in various tissues of patients with neurodegenerative disease. Here we studied the protein levels of complex I 24- and 75-kDa subunits in several brain regions from patients with Down syndrome (DS) and Alzheimer's disease (AD). We determined protein levels of complex I 24-, 75-kDa subunits and mitochondrial marker proteins mitochondrial matrix protein P1 (hsp60) and aconitate hydratase from seven brain regions of patients with DS, AD and controls. Proteins were separated by two-dimensional (2-D) gel electrophoresis and identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). Complex I 24-kDa subunit was significantly reduced in occipital cortex and thalamus in patients with DS and temporal and occipital cortices in patients with AD. Complex I 75-kDa subunit was significantly reduced in brain regions from patients with DS (temporal, occipital and caudate nucleus) and AD (parietal cortex). Reductions of two subunits of complex I may lead to the impairment of energy metabolism and result in neuronal cell death (apoptosis), a hallmark of both neurodegenerative disorders. PMID- 11400917 TI - Significance of GABAergic systems in the action of improgan, a non-opioid analgesic. AB - Improgan is the prototype drug from a new class of non-opioid analgesics chemically related to histamine and histamine antagonists, but the mechanism of action of these compounds has not been identified. Because several classes of analgesics act in the brain by reducing GABAergic inhibition of endogenous pain relieving circuits, and because the activity of these substances is abolished by the GABA(A) agonist muscimol, the present study assessed the effects of muscimol on improgan antinociception in rats. Intracerebroventricular (icv) improgan (80 microg) and morphine (20 microg) both induced 80-100% of maximal analgesic responses on the tail flick test 10 to 30 min later. However, muscimol pretreatment (0.5 microg, icv) completely eliminated the antinociceptive activity of both compounds. Since improgan in vitro lacks activity at opioid and GABA(A) receptors, these findings: 1) confirm earlier literature showing that muscimol inhibits morphine analgesia, and 2) suggest that improgan activates a supraspinal, descending analgesic pathway, possibly via inhibition of GABAergic transmission. Since muscimol is the first compound discovered which inhibits improgan analgesia, muscimol will be a useful tool for the further characterization of this new class of pain-relieving substances. PMID- 11400918 TI - Evidence that L-deprenyl treatment for one week does not inhibit MAO A or the dopamine transporter in the human brain. AB - In this study, we investigated whether treatment with L-deprenyl, a selective monoamine oxidase B (MAO B) inhibitor, also inhibits MAO A or the dopamine transporter in the human brain. Six normal volunteers (age 46+/-6 yrs) had two PET sessions, one at baseline and one following L-deprenyl (10 mg/day) for 1 week. Each session included one scan with [11C]clorgyline (to assess MAO A) and one scan 2 hours later with [11C]cocaine (to assess dopamine transporter availability). A 3-compartment model was used to compare the plasma-to-brain transfer constant, K1 (a function of blood flow) and lambdak3 (a kinetic term proportional to brain MAO A) before and after treatment. Dopamine transporter availability was measured as the ratio of distribution volumes of the striatum to cerebellum (DVR) which is equal to Bmax/KD +1. L-Deprenyl treatment for 1 week did not affect either brain MAO A activity or dopamine transporter availability. There was a non-significant trend for an increase in K1 after L-deprenyl. These results confirm that L-deprenyl after one week of treatment at doses typically used clinically is selective for MAO B and that it does not produce a measurable affect on the dopamine transporter, suggesting that MAO A inhibition and dopamine transporter blockade do not contribute to its pharmacological effects. PMID- 11400919 TI - Prostate specific antigen and screening for early prostate cancer. PMID- 11400920 TI - Follow-up after attempted curative surgery for colorectal cancer; postal survey of New Zealand surgeons' practice. AB - AIMS: The role of follow-up after attempted curative resection of colorectal cancer (CRC) is not clearly defined. We wished to establish the frequency, duration and type of follow-up practised by New Zealand surgeons. METHODS: A postal survey was performed of surgeons on the General Medical Register, asking about the follow-up of asymptomatic patients after potential curative surgery. RESULTS: The response rate was 66%(107/163). There was wide variability in the frequency, duration and type of the indicated follow-up practice. 97% of surgeons followed their patients on average four monthly for the first year. At five years, 79% of surgeons followed their patients. Routine blood tests were performed 54%, while serum carcinoembryonic antigen (CEA) levels were measured by 56% of surgeons for the first two years. 41% performed abdominal imaging in the first two years. 97% of surgeons screened the remaining colon (88% by colonoscopy). 90% performed colonic screening three to five yearly. CONCLUSIONS: Follow-up after potential curative surgery for CRC appears to be widely practised in New Zealand. There is, however, considerable variation between surgeons in the frequency, duration and type of follow-up. This may reflect the conflicting evidence in the literature on the value of follow-up. The outcome of current large prospective randomised trials may confirm whether or not such follow-up is worthwhile and what form it should take. PMID- 11400921 TI - Hip fracture incidence in New Zealand, revisited. AB - AIM: Earlier predictions of the incidence of hip fractures in the older adult population suggested that by 2011 the rate would rise to epidemic proportions. The purpose of this study was to compare the number hip fractures occuring in New Zealand from 1988 to 1999 with the hip fracture rate predicted in 1990 by Rockwood, Horne and Cryer. METHODS: Data on the number of patients admitted to New Zealand hospitals with a diagnosis of fractured neck of femur were obtained, and compared with weighted regression and baseline predictions of Rockwood et al. RESULTS: The numbers of hip fractures for females, from 1988 to 1993, were similar to the number predicted, yet have been significantly lower than stated predictions since 1995. For males, hip fracture numbers since 1995 were less than the weighted regressions predicted (NS). CONCLUSIONS: Numbers of hip fractures since 1995 have been fewer than predicted. Possible reasons for maintaining the rates of hospitalisation due to fractured neck of femur at pre-1995 levels, are discussed. PMID- 11400922 TI - The epidemiology of emergency department attendances in Christchurch. AB - AIM: To describe the characteristics of patients who attended Christchurch Hospital's emergency department (ED) in 1998. METHODS: Non-identifiable data about all the attendances to the ED during 1998 were obtained from Christchurch Hospital's management information services database. Demographic data, time and date of arrival, source of referral, diagnosis, triage category, and discharge destination were analysed. RESULTS: In 1998 there were 65,024 attendances, on average 178 people per day (95% confidence interval 145-211). Children and the elderly were the most frequent attenders except in weekends when visits by young people were more common. The elderly had the highest age specific rates of attendance. Most attendances were between 8am and midnight, and Mondays and July were the day and month with most attendances respectively. 43% of people referred themselves and 38% were referred by their general practitioner (GP). GP referrals were significantly more common during the day, the working week and in the winter. The elderly frequently presented with medical problems, their attendances were classified as more urgent and 75% of their visits resulted in hospital admission. CONCLUSIONS: These findings are generally consistent with the few previously published descriptions of ED attendance in New Zealand and most overseas studies. ED attendance data have important implications for funders and providers of ED services and a national dataset should be established. PMID- 11400923 TI - Frequent attenders at Christchurch Hospital's emergency department. AB - AIM: To describe the characteristics of adult frequent attenders presenting to a major urban emergency department during a twelve month period. METHODS: A retrospective study was performed of all frequent attenders (presenting ten of more times during twelve months) to the Emergency Department (ED) at Christchurch Hospital. RESULTS: The proportion of visits to the ED by frequent attenders increased from 1.38% to 2.08% of all visits between 1996 to 2000. A computer search identified 86 frequent attenders between 1 November 1998 and 31 October 1999: 1,395 visits were made by these patients. The median age was 35 years, both sexes were equally represented and most frequent attenders were single. The most common presenting problem was medical (40%) or psychological (36%), and (86%) had secondary problems related to each attendance. CONCLUSION: This study's findings are generally consistent with those from two previous descriptions of frequent attenders. Evidence from overseas trials suggests that case management may provide the best outcomes for these patients, but further research is needed. PMID- 11400924 TI - Medical care at the Sweetwaters Music Festival. AB - AIMS: To describe medical cover and medical presentations at the four-day 1999 Sweetwaters Music Festival, and make comparisons with other festivals. METHODS: All medical contacts were counted, and patients presenting to the medical tent were included in the study. Case records were studied to determine demographic data, nature of complaint, treatment and disposition. A Medline literature search was performed to obtain information on other festivals. RESULTS: There were 2,231 medical contacts overall (8.9% of estimated attendees) and 217 presentations to the medical tent (0.9% of estimated attendees). 53% of patients presenting to the medical tent were men and the mean patient age was 25 years. Lacerations (16%), intoxication (13%), local infections (12%) and soft tissue injuries (9%) were the most common problems. There were no deaths or cardiac arrests. CONCLUSION: Problems encountered were similar to other music festivals, with minor injuries predominant. PMID- 11400925 TI - Morbidity associated with gastrostomy placement in children demands an ongoing integrated approach to care. AB - AIMS: To evaluate the short and long term morbidity of gastrostomy insertion, and to identify ongoing management requirements. METHODS: A retrospective review was undertaken of the hospital casenotes of children aged up to fifteen years who had a gastrostomy placed in Christchurch over a six year period to March 1998. RESULTS: 42 children had a gastrostomy fashioned, 35 in the last three years of the period reviewed. The most common underlying diagnosis was neurological disease (48%), and the most common indication for tube placement was failure to feed orally. Complications were frequent but minor. Morbidity was related to local erythema and infection around the stoma (85 episodes in 23 children), persistent and major gastric fluid leakage (three episodes), and mechanical failure of the tube (21 episodes). Gastro-oesophageal reflux was seen in fourteen children, nine of whom had primary neurological disease. Complications were seen more after open gastrostomy than after percutaneous endoscopic placement (6.6:4.7). There was no mortality related directly to the gastrostomy tube or tube placement. CONCLUSIONS: An increase in the frequency of gastrostomy placements has been seen over this period. As the number of children with a gastrostomy increases, so too have the demands on medical and nursing staff to care for and manage the devices. The frequency of minor ongoing problems necessitates ongoing support of the child and care of the gastrostomy. A close working relationship between outreach nursing staff, stoma therapists and medical staff is required if morbidity is to be minimised. Education, audit and review remain important additional aspects of care. PMID- 11400926 TI - The competence and performance of medical practitioners. PMID- 11400927 TI - Obesity, oral contraceptives, and fatal pulmonary embolism. PMID- 11400928 TI - Tobacco and taking responsibility. PMID- 11400929 TI - The genetic sonogram in screening for Down syndrome: response to the JAMA study. PMID- 11400930 TI - Use of genetic sonography in screening for trisomy 21. PMID- 11400931 TI - Importance of evaluating organ parenchyma during screening abdominal ultrasonography after blunt trauma. AB - OBJECTIVE: To determine the benefit of screening ultrasonography for parenchymal abnormalities as well as free fluid during screening abdominal ultrasonography in patients with blunt trauma. METHODS: A total of 2693 patients with blunt trauma who were triaged to a level 1 trauma center underwent screening abdominal ultrasonography in the resuscitation suite. Examinations were performed by experienced sonographers and included a screen for free intraperitoneal fluid and evaluation of the abdominal organ parenchyma and heart for traumatic injury. Screening ultrasonographic findings were reviewed and compared with findings from autopsy, laparotomy, diagnostic peritoneal lavage, computed tomography, repeated ultrasonography, cystography, and clinical outcome. Imaging studies of all patients with confirmed or suspected injuries were reviewed to identify those in whom parenchymal findings aided diagnosis. RESULTS: One hundred seventy-two patients were found to have evidence of abdominal injury due to blunt trauma on the basis of clinical data, imaging, laparotomy, or autopsy. Forty-four of these patients had no sonographic evidence of hemoperitoneum at the time of initial ultrasonography. Screening ultrasonographic findings were positive for injury in 19 of 44 patients on the basis of parenchymal findings or small retroperitoneal collections of fluid thought to be indicative of trauma. In the remaining 25 patients, screening ultrasonography showed no abnormalities, and injuries were detected by repeated ultrasonography, subsequent computed tomography, or diagnostic peritoneal lavage performed for suspected occult injury on the basis of clinical parameters. In addition, 47 of 126 injured patients with sonographically detected free fluid had parenchymal findings that helped localize injury. Sixteen of those patients were taken to the operating room on the basis of clinical and sonographic findings without undergoing computed tomography. CONCLUSIONS: The inability to show injuries with no hemoperitoneum or with delayed hemoperitoneum has been shown to be a limitation of ultrasonography in patients with blunt trauma. In our series, 26% of all patients with documented injuries had no free fluid visible on screening ultrasonography Attention to findings other than free fluid allowed detection in 43% of injured patients without sonographic evidence of hemoperitoneum. PMID- 11400932 TI - Sonographic diagnosis of distal biceps tendon rupture: a prospective study of 25 cases. AB - OBJECTIVE: To describe the sonographic findings of distal biceps tendon ruptures and to assess the accuracy of the technique. METHODS: Twenty-five patients with clinically indicated distal biceps tendon ruptures were prospectively studied by sonography. Five patients also underwent magnetic resonance imaging. Surgical confirmation was obtained in 14 patients. RESULTS: Seventeen complete tendon ruptures and 3 partial tears were correctly shown by sonography; 1 complete rupture was incorrectly shown as a partial tear by sonography. Sonographic features of complete rupture were absence of tendon in the expected location, fluid collection in a typical tendon gap, and a mass in the antecubital fossa. Sonographic features of incomplete rupture were intratendinous hypoechogenicity and tendon thinning. Peritendinous fluid was found in complete and incomplete ruptures. CONCLUSIONS: In distal biceps tendon ruptures, sonography is a cost effective method that can confirm the clinical indications with good accuracy and can show tendon lesions when the clinical indications are low. PMID- 11400933 TI - Ultrasonographic evaluation of diaphragmatic motion. AB - OBJECTIVE: To evaluate the technical feasibility and utility of ultrasonography in the study of diaphragmatic motion at our institution. METHODS: The study consisted of 2 parts. For part I, in 23 volunteers we performed 23 studies on 46 hemidiaphragms with excursions documented on M-mode ultrasonography For part II, in 22 patients we performed 52 studies in 102 hemidiaphragms. In 50 studies both hemidiaphragms were studied, and in another 2 studies only 1 hemidiaphragm was studied. Patients' ages ranged from birth to 66 years (mean, 23 years). There were 16 male and 6 female patients. Indications for the study were (1) suggestion of paralysis of the diaphragm (n = 22); (2) if the diaphragm was already known to be paralyzed, for evaluation of response to phrenic nerve or pacer stimulation (n = 9); and (3) follow-up of previous findings (n = 21). Patients were examined in the supine position in the longitudinal semicoronal plane from a subcostal or low intercostal approach. Motion was documented with real-time ultrasonography and measured with M-mode ultrasonography. RESULTS: Of the 102 clinical hemidiaphragms studied, findings included normal motion (n = 42), decreased motion (n = 22), no motion (n = 6), paradoxical motion (n = 10), positive pacer response (n = 13), negative pacer response (n = 2), positive phrenic stimulation (n = 6), and negative phrenic stimulation (n = 1). There were no failures of visualization. CONCLUSIONS: Ultrasonography proved feasible and useful in evaluating diaphragmatic motion. In our practice it has replaced fluoroscopy. Ultrasonography has advantages over traditional fluoroscopy, including portability, lack of ionizing radiation, visualization of structures of the thoracic bases and upper abdomen, and the ability to quantify diaphragmatic motion. PMID- 11400934 TI - Quantifying B-mode images of in vivo rat mammary tumors by the frequency dependence of backscatter. AB - OBJECTIVE: To evaluate the frequency dependence of ultrasonic backscatter for its ability to differentiate between neoplastic and healthy tissue. METHODS: Standard B-mode images were created of 5 rats with spontaneous mammary tumors, and regions of interest in the lesion and surrounding tissue were parameterized by the slope of the backscatter amplitude versus frequency. RESULTS: In 4 of the 5 rats, the averaged backscatter slope of the regions of interest in the tumor was significantly (P < .05) different from that of the surrounding tissue, and the fifth case had a moderate difference (P = .20). The consistency of the averaged slope values (1.2-1.8 dB/MHz) across all but 1 of the mammary tumors was encouraging for the prospect of identifying a tissue type by its backscatter slope. CONCLUSIONS: This work suggests that characterization and diagnosis of tissue types may be possible by using ultrasonographic images quantified by the frequency dependence of backscatter. PMID- 11400935 TI - Ultrasonography of the fetal thyroid: nomograms based on biparietal diameter and gestational age. AB - OBJECTIVE: To describe gestational age-dependent and -independent nomograms for fetal thyroid size. METHODS: Two hundred fetuses were evaluated between 16 and 37 weeks' gestation in this cross-sectional study. RESULTS: Nomograms of fetal thyroid size were created by using the 5th, 10th, 50th, 90th, and 95th percentiles based on biparietal diameter and gestational age. A second-order polynomial fit for biparietal diameter and a linear fit for gestational age best described thyroid circumference measurements. Variations in thyroid circumference measurements increased with both larger biparietal diameter and advancing gestational age. There was no intraobserver or interobserver variability in thyroid circumference measurements (P > .20). CONCLUSIONS: Both biparietal diameter and gestational age serve as good predictors of fetal thyroid circumference. When the biparietal diameter is difficult to measure, gestational age can be used to assess thyroid size. PMID- 11400936 TI - The single umbilical artery in a high-risk patient population: what should be offered? AB - OBJECTIVE: To determine whether fetal echocardiography is warranted in cases of single umbilical artery in a population at risk for aneuploidy. METHODS: All cases of fetal single umbilical artery identified over a 2-year period were reviewed for other sonographically detected abnormalities, fetal echocardiographic results, and karyotype. RESULTS: Sixty-five cases of single umbilical artery were diagnosed on the basis of initial sonograms. Five were subsequently shown to have 3-vessel cords (8% false-positive diagnosis; incidence, 1.2%). Excluding 3 from twin gestations, 57 cases formed the study population. Thirty-one fetuses (54%) were initially thought to have isolated single umbilical arteries, and 26 (46%) had nonisolated single umbilical arteries. Fetal echocardiography was performed in 29 cases (51%), 24 (83%) with normal findings and 5 (17%) with abnormal findings. Four (50%) of 8 nonisolated single umbilical arteries had abnormal echocardiographic findings versus 1 (5%) of 21 apparently isolated single umbilical arteries (P < .05; odds ratio, 20). Karyotypes in 36 cases (63%) showed 25 (69%) euploid and 11 (31%) aneuploid fetuses. An apparently isolated single umbilical artery was never associated with an abnormal karyotype. Eleven (50%) of 22 fetuses with nonisolated single umbilical arteries had aneuploidy (P < .005). The side of the missing umbilical artery did not correlate with other sonographically detected abnormalities, abnormal fetal echocardiographic findings, or aneuploidy. CONCLUSIONS: The rate of cardiac malformations seen with apparently isolated single umbilical arteries is significant, and fetal echocardiography should be performed. PMID- 11400937 TI - A reappraisal of amniotic fluid alpha-fetoprotein measurement at the time of genetic amniocentesis and midtrimester ultrasonography. AB - OBJECTIVE: To evaluate the contemporary utility of amniotic fluid alpha fetoprotein measurement as a complementary test for fetal abnormalities at the time of invasive genetic testing. METHODS: A review of amniotic fluid alpha fetoprotein test results was conducted to determine the frequency with which elevated alpha-fetoprotein values added independent diagnostic information and altered clinical management. Amniotic fluid specimens processed for alpha fetoprotein between 1995 and 1998 were included. Elevated alpha-fetoprotein cases were classified as either incidental to the fetal abnormality diagnosed or central to the identification of a fetal anomaly on the basis of whether an ultrasonographic examination had already identified the anomaly before amniocentesis. The costs associated with alpha-fetoprotein testing were used to estimate the expenditure per pregnancy in which elevated alpha-fetoprotein values would add discriminatory diagnostic value. A hypothetical national cost model was constructed to explore the utility of selective rather than routine amniotic fluid alpha-fetoprotein measurement. RESULTS: Eighty-two (3%) of 2769 amniotic fluid alpha-fetoprotein values were elevated. In only 1 instance was the elevated result found to be partially discriminatory (e.g., an established diagnosis of microcephaly with an associated small encephalocele identified after the elevated amniotic fluid alpha-fetoprotein value prompted repeated ultrasonographic assessment). Sixty-one other neural tube defects were detected by ultrasonography alone (myelomeningocele, n = 28; anencephaly, n = 24; and encephalocele, n = 9). Thus, an elevated alpha-fetoprotein result added diagnostic precision in only 1 (0.036%) of 2769 cases. Cost estimates suggested that routine amniotic fluid alpha-fetoprotein assessment resulted in a $219,000 expenditure per informative case. CONCLUSIONS: Routine measurement of amniotic fluid alpha-fetoprotein during amniocentesis may not be warranted in centers with expertise in targeted ultrasonographic imaging. PMID- 11400938 TI - Normal midtrimester (17-20 weeks) genetic sonogram decreases amniocentesis rate in a high-risk population. AB - OBJECTIVE: To evaluate a screening protocol using advanced maternal age, triple marker screening, and genetic sonography. METHODS: We compared adverse chromosomal outcomes of pregnancy in 1556 women referred for increased risk of aneuploidy because of either advanced maternal age or triple-marker test results. Patients were counseled about the results of the triple-marker test and subsequent sonography, which led to a patient decision of whether to pursue amniocentesis. Fetal measurements and structural abnormalities were compared with chromosomal findings. When patients elected amniocentesis, karyotypes were obtained. RESULTS: Genetic sonography reduced the rate of amniocentesis by 61% overall and by 40% when compared with an alpha-fetoprotein profile alone. The sensitivity of sonography combined with the triple-marker screen for the detection of trisomy 21 was 87% compared with 91% for the triple-marker screen alone. CONCLUSIONS: This study confirmed that sonographic findings in a targeted population, in combination with other risk markers (advanced maternal age and triple-marker screening), can be used to assess the risk of aneuploidy. Biometry provides additional information for assessing the risk of aneuploidy. Combining advanced maternal age, serum triple-marker screening, and sonographic screening may provide better risk prediction for use in clinical counseling. PMID- 11400939 TI - Combined use of genetic sonography and maternal serum triple-marker screening: an effective method for increasing the detection of trisomy 21 in women younger than 35 years. AB - OBJECTIVE: The current standard of practice is to screen women younger than 35 years for trisomy 21 with maternal triple-marker screening, followed by amniocentesis in high-risk (1:10-1:190) patients. Non-high-risk patients are not offered further diagnostic testing. This study was conducted to determine whether genetic sonography of fetuses considered to be at moderate risk (1:190-1:1000) after maternal triple-marker screening increases the detection for trisomy 21, is cost-effective, and reduces the number of amniocenteses required to detect a single fetus with trisomy 21. METHODS: After triple-marker screening, mathematical modeling was used to classify 500,000 theoretical fetuses as high, moderate, or low risk (>1:1001-1:10,000) for trisomy 21. The sensitivity for genetic sonography varied between 60% and 90%, and false-positive rates varied between 5% and 25%. Two programs (I and II) were compared with the control program. The control program included patients with high-risk fetuses (1:10 1:190) who had amniocentesis. Program I consisted of patients in the moderate risk group (1:191-1:1000) who had genetic sonography followed by amniocentesis only when an abnormal sonographic finding was present. Program II used an approach in which genetic sonography was done for both high- and moderate-risk fetuses, and amniocentesis was performed only when an abnormal sonographic finding was present. RESULTS: When added to the control program, genetic sonography significantly increased the detection rate of Down syndrome (range, 68.1%-77.8% versus 49%), reduced the cost of detection, and resulted in a ratio of fetuses with Down syndrome detected to normal fetuses lost because of amniocentesis of greater than 1 (range, 2.1-4.2). Compared with the control program, program II significantly increased the detection of fetuses with trisomy 21 (range, 56%-72%) when the sensitivity of genetic sonography was >70%, reduced the cost of detection, and had ratios of trisomy 21 detection to normal fetuses lost because of amniocentesis of between 2.38 and 17.88. CONCLUSION: Women younger than 35 years and classified as having moderate risk after triple-marker screening could undergo genetic sonography under 1 of 2 approaches, either of which would result in an increased detection rate of trisomy 21 and be cost effective without increasing the loss rate of normal fetuses after genetic amniocentesis. PMID- 11400940 TI - Sonographic markers of fetal trisomies: second trimester. AB - OBJECTIVE: Second-trimester sonographic findings of fetal trisomy may include structural abnormalities or sonographic markers of fetal aneuploidy. Unlike structural anomalies, sonographic markers of fetal aneuploidy are insignificant by themselves with regard to outcome, are nonspecific--most frequently seen in normal fetuses, and are often transient. Our objective was to review the second trimester sonographic findings of the major trisomic conditions, trisomies 13, 18, and 21. METHODS: We reviewed a number of the most commonly accepted markers, including nuchal thickening, hyperechoic bowel, echogenic intracardiac focus, renal pyelectasis, shortened extremities, mild cerebral ventricular dilatation, and choroid plexus cysts. Markers associated with trisomy 21 were emphasized. RESULTS: The sensitivity of sonography for detection of fetal trisomic conditions varies with the type of chromosome abnormality, gestational age at the time of sonography, reasons for referral, criteria for positive sonographic findings, and the quality of the sonography. As an estimate, 1 or more sonographic findings can be identified in approximately 90% of fetuses with trisomy 13, 80% of fetuses with trisomy 18, and 50% to 70% of fetuses with trisomy 21 (Down syndrome). CONCLUSIONS: The presence or absence of sonographic markers can substantially modify the risk of fetal Down syndrome and is the basis of the so-called genetic sonogram. Because maternal biochemical and sonographic markers are largely independent, combined risk estimates will result in even higher detection rates than either alone. PMID- 11400941 TI - Renal Doppler ultrasonographic findings in earthquake victims with crush injury. AB - OBJECTIVE: In the Marmara earthquake, which occurred on August 17, 1999, 639 people had renal complications, and 477 needed hemodialysis treatment because of acute renal failure due to crush injury. Our objective was to use Doppler ultrasonography as a noninvasive procedure to provide renal hemodynamic information. METHODS: We evaluated 9 patients with severe crush injury by using Doppler ultrasonography to measure renal resistive indexes within 16 to 32 hours after the earthquake and 6 weeks later. All patients had acute renal failure and underwent hemodialysis. RESULTS: All measured initial resistive indexes were high (mean +/- SD, 0.83 +/- 0.08), and they were correlated with the number of hemodialysis sessions (r = 0.703; P = .034), the duration of hemodialysis treatment (r = 0.819; P = .007), and oligoanuria (r = 0.937; P < .0001). All patients recovered from the acute renal failure, and repeated Doppler ultrasonography showed significant decreases in resistive indexes (0.59 +/- 0.07; P < .01) and renal lengths (P < .05). CONCLUSIONS: In crush injury, measurement of renal resistive indexes can be useful for the prognosis of recovery from acute renal failure. PMID- 11400942 TI - Imaging of the ductus venosus in neonates: from patency to closure. AB - OBJECTIVE: To familiarize the radiologist with the variable sonographic appearance of the involuting ductus venosus in neonates. METHODS: Five sick neonates ranging in age from 24 to 42 weeks had abdominal sonographic examinations to check for intra-abdominal diseases. RESULTS: Doppler sonography showed a patent ductus venosus in 3 neonates and a ductus venosus with thrombosis in 2 neonates. The ductus venosus was identified in the liver between the left portal vein and the inferior vena cava. In 2 patients, follow-up sonography showed that the ductus venosus progressively had thrombosis and then disappeared. CONCLUSIONS: The ductus venosus in a neonate has a variable sonographic appearance depending on its stage of involution. Its detection in a sick neonate can lead to inadvertent diagnosis of a hepatic varix, an abscess, or a tumor. PMID- 11400943 TI - Color Doppler ultrasonography of the superior mesenteric artery for prenatal ultrasonographic diagnosis of a left-sided congenital diaphragmatic hernia. AB - The incidence of congenital diaphragmatic hernia (CDH) has been estimated as 1 per 2000 to 1 per 4000 births. The etiology of the malformation is unknown, but it has been reported in association with maternal administration of medications such as thalidomide or antiepileptics before closure of the pleuroperitoneal canal at 9 to 10 weeks' gestation as well as having a familial inheritance pattern. Congenital diaphragmatic hernia is associated with other congenital anomalies in 25% to 57% of cases and with chromosomal abnormalities in 10% to 20% of cases. Posterolateral, anterolateral, and pars sternalis defects of closure of the pleuroperitoneal canal encompass the 3 types of CDH. The most frequent type is the left-sided posterolateral defect or Bochdalek's hernia, which accounts for 81% of cases. PMID- 11400944 TI - Prenatal ultrasonographic diagnosis of Sprengel's deformity. AB - Sprengel's deformity is the congenital failure of descent of the scapula. Eulenburg first described this in 1863. Sprengel described 4 cases in 1891; hence, the anomaly has been called Sprengel's deformity. Other terms for this entity are Sprengel's anomaly, Sprengel's shoulder, congenital high scapula, and undescended scapula. In this entity, varying degrees of scapula elevation and scapula hypoplasia occur. In approximately one third of cases, Sprengel's deformity is accompanied by an accessory ossicle, the omovertebral bone, which articulates between the medial border of the scapula and 1 or more of the cervical vertebrae. In the absence of an omovertebral bone, a fibrous fascial sheath extends from the superior angle of the scapula to the spinous process, the lamina, or the transverse process of 1 or more lower cervical vertebrae. This fixation accounts for the medial position and elevation of the scapula. I report a case of prenatal ultrasonographic detection of Sprengel's deformity. PMID- 11400945 TI - A case of chondrodysplasia punctata with features of osteogenesis imperfecta type II. AB - The osteogenesis imperfecta syndromes constitute a group of heterogeneous, heritable skeletal dysplasias. Of the 4 types, type II is the most severe, with an incidence of 1 per 55,000. It is characterized by malformed bones secondary to abnormal collagen type I synthesis. Affected fetuses are divided into 3 groups: A, B, and C. All groups have long bones described as "wrinkled" or "crumpled" secondary to repeated fractures. Many bones also show evidence of demineralization, which is especially evident in the bones of the face and calvaria. In groups A and C, the chest is generally small, with thickened and shortened ribs, and each rib has characteristic "beading" patterns secondary to repeated fracturing. Sonography has traditionally been successful in the diagnosis of osteogenesis imperfecta at an early gestational age. Chondrodysplasia punctata describes a heterogeneous group of skeletal disorders characterized by abnormal mineralization of bones during gestation. There are many different causes of it, but some of the specific subtypes include rhizomelic, X-linked dominant (also known as Conradi-Hunermann syndrome), X linked recessive, and tibia-metacarpal. We report a case of severe X-linked dominant chondrodysplasia punctata, which sonographically had common features with osteogenesis imperfecta type II. PMID- 11400946 TI - Cytomegalovirus-specific cytolytic T-cell lines and clones generated against adenovirus-pp65-infected dendritic cells. AB - Cytomegalovirus (CMV) infection is a serious complication of allogeneic bone marrow transplantation (BMT). CMV disease can usually be prevented by passive immunization with donor-derived CMV-pp65-specific T-cell clones if provided early post-BMT. The classic method of generating CMV-specific T-cell clones requires donor-derived fibroblast lines infected with CMV as stimulators, thus limiting the availability of CMV immunotherapy to those patients for whom a donor skin biopsy can be obtained 6 to 8 weeks pretransplantation. To overcome this limitation we have used monocyte-derived dendritic cells (DCs) to induce donor anti-CMV cytotoxic T lymphocytes (CTLs). Matured, adeno-pp65-infected DCs were added at day 0 and at day 7 of a 2-week culture of donor peripheral blood mononuclear cells. DC-primed cultures were compared with cultures stimulated in an identical fashion with CMV-infected fibroblasts or with adeno-pp65-infected freshly isolated blood monocytes. Specific killing of CMV-infected fibroblasts was detected in all except the culture stimulated with pp65-infected monocytes. DCs infected after maturation elicited greater CTL activity than did DCs matured after infection. A series of 5 CD8+ clones from a fibroblast-stimulated culture and 7 CD8+ clones from a mature-DC-stimulated culture derived from a single HLA A*0201+ individual were characterized. All 12 clones lysed autologous CMV infected fibroblasts. All except 1 clone from the CMV-infected fibroblast arm (fibroblast arm) lysed vaccinia-pp65-infected B-lymphoblastoid cell lines (BLCLs); none lysed vaccinia-pp150-infected or noninfected BLCLs. Ten of 10 CD8+ clones tested were restricted by HLA-A*0201. Seven of the 12 clones were Vbeta6+ (2 from the fibroblast arm and 5 from the DC arm) with an identical Vbeta6.1-J1.4 sequence. Three clones from the fibroblast arm and 5 clones from the DC arm recognized the pp65 peptide NLVPMVATV (amino acids [aa], 495-503). These data show that CMV-specific T-cell clones with similar restriction patterns, T cell receptor usage, and specificity can be generated using monocyte-derived pp65 infected-DC or CMV-infected-fibroblast stimulators. This approach should broaden the applicability of CMV-specific T-cell immunotherapy to a wider spectrum of patients by reducing the time required to generate CMV-specific T-cell clones. PMID- 11400947 TI - Bone density loss after allogeneic hematopoietic stem cell transplantation: a prospective study. AB - The incidence and course of bone density abnormalities following hematopoietic stem cell transplantation are poorly understood and complicated by the impact of multiple factors. Hip, spine, and wrist bone mineral densities (BMDs) were measured in 104 adults (54 women, 54 men; mean age, 40 years [range, 18-64 years]) at 3 and 12 months after allogeneic transplantation. Clinical and laboratory variables were evaluated using univariate and multivariate analyses to determine risk factors for osteoporosis, fracture, and avascular necrosis. At 3 months posttransplantation, combined (male and female) hip, spine, and wrist z scores were -0.35, -0.42, and +0.04 standard deviations, respectively. At 12 months both men and women experienced significant loss of hip BMD (4.2%, P < .0001); changes in the spine and wrist were minimal. The cumulative dose and number of days of glucocorticoid therapy and the number of days of cyclosporine or tacrolimus therapy showed significant associations with loss of BMD; age, total body irradiation, diagnosis, and donor type did not. Nontraumatic fractures occurred in 10.6% of patients and avascular necrosis in 9.6% within 3 years posttransplantation. The decrease in height between pretransplantation and 12 months posttransplantation was significant (P = .0001). Results indicate that loss of BMD after allogeneic stem cell transplantation is common and accelerated by the length of immunosuppressive therapy and cumulative dose of glucocorticoid. An increased incidence of fracture and avascular necrosis may adversely impact long-term quality of life. Prevention of bone demineralization appears warranted after stem cell transplantation. PMID- 11400948 TI - Randomized clinical trial of thalidomide, cyclosporine, and prednisone versus cyclosporine and prednisone as initial therapy for chronic graft-versus-host disease. AB - Chronic graft-versus-host disease (CGVHD) is a major cause of morbidity following allogeneic bone marrow transplantation. Thalidomide is active in salvage therapy for high-risk or resistant CGVHD. In a prospective randomized trial, we tested initial therapy with thalidomide. Patients with extensive CGVHD were randomized to receive either cyclosporine and alternate-day prednisone (n = 27, no thalidomide [no-thal] group) or cyclosporine, prednisone, and thalidomide (200 800 mg/day; n = 27, thal group). Although most patients responded, initial therapy with thalidomide did not improve control of CGVHD. Response rates were 83% versus 89% at 2 months (P = .7), 88% versus 84% at 6 months (P > .8) and 85% versus 73% at 1 year (P = .5) in the thal and no-thal groups, respectively. Multivariate analysis revealed related donor transplant (odds ratio [OR] = 11.3; P =.03) and de novo or quiescent onset of CGVHD (OR = 7.7; P =.04) to be significant predictors of good early response, whereas a platelet count of > or =100,000/microL was a significant predictor of good response (OR = 10.4; P =.04) at 1 year. Survival for the thal and no-thal groups was similar at 1 year (66% versus 74%) and 2 years (66% versus 54%, P = .85). Multivariate analysis revealed progressive onset CGVHD (relative risk [RR] = 4.2; P =.01), unrelated donor (RR = 5.7; P < .01), sex mismatch (RR = 7.9; P < .01), and platelet counts of <100,000/microL (RR = 3.8; P = .01) as significant predictors of poorer survival. These data suggest that despite a high response rate (79% response and 53% complete response) and encouraging survival rates (70% at 1 year and 60% at 2 years), thalidomide offers no clinical benefit when incorporated into initial therapy for CGVHD. The value of thalidomide as salvage therapy requires further study. PMID- 11400949 TI - Inhaled steroids as prophylaxis for delayed pulmonary toxicity syndrome in breast cancer patients undergoing high-dose chemotherapy and autologous stem cell transplantation. AB - PURPOSE: To evaluate the efficacy of inhaled fluticasone propionate (Flovent) as prophylaxis against delayed pulmonary toxicity syndrome (DPTS) and decline in pulmonary function in breast cancer patients undergoing high-dose chemotherapy with the conditioning regimen of cyclophosphamide, cisplatin, and carmustine (CPB) followed by autologous stem cell transplantation (ASCT). PATIENTS AND METHODS: Sixty-three consecutive patients with multinode-positive or metastatic breast cancer undergoing high-dose chemotherapy with CPB and ASCT who were treated at the Duke University Adult Bone Marrow Transplant Program. All patients were started on inhaled fluticasone propionate, 880 microg every 12 hours, for 12 weeks from the start date of their CPB conditioning regimen. Pulmonary function tests (PFTs) with a single-breath diffusing capacity of carbon monoxide (DLCO) were performed pre-ASCT as well as approximately 6 and 12 weeks post-ASCT. DPTS was defined as follows: (1) development of a nonproductive cough and dyspnea with or without fever, plus a fall in DLCO to less than 60% predicted; or (2) decline in DLCO to less than 50% predicted with or without symptoms. RESULTS: Pulmonary function tests were done on all patients pre-ASCT, on 56 of the 63 patients at a median of 44 days (range, 25 to 73 days) post-ASCT, and on 51 of the 63 patients at a median of 96 days (range, 50 to 190 days) post-ASCT. The PFTs showed an average of an 8% (+/-26%) and 21% (+/-22%) decline in DLCO. These declines compare favorably with our historical control group of 45 consecutive breast cancer patients undergoing ASCT with CPB as a conditioning regimen, who experienced average declines in DLCO of 29% (+/-18%) (P < .001) and 33% (+/-18%) (P < .001) at comparable time periods post-ASCT. Delayed pulmonary toxicity syndrome occurred in 35% of treated patients compared to 73% of the historical controls (P = .0003). No patients died of DPTS or pulmonary problems, and there were no fungal pneumonias. CONCLUSION: Inhaled fluticasone propionate may decrease the incidence of DPTS in patients treated with CPB as a conditioning regimen for ASCT, as well as help to preserve pulmonary function as measured by DLCO. These results are worthy of further study in a randomized clinical trial. PMID- 11400950 TI - Male sexual function after autologous blood or marrow transplantation. AB - Although endocrine dysfunction has been reported in survivors of allogeneic bone marrow transplantation (alloBMT), data for autologous BMT (autoBMT) recipients are lacking. Because information on male potency in particular is scanty, we prospectively assessed male sexual function after autoBMT. We identified 16 men who were < or =50 years of age at the time of evaluation and disease free for at least 6 months after autoBMT. Nine had Hodgkin's disease, 4 had acute myelogenous leukemia, and 3 had non-Hodgkin's lymphoma. Blood samples were assayed for luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone. Patients were surveyed with a modified version of the Pyschosocial Adjustment to Illness Scale regarding erectile dysfunction and loss of interest in sexual activities. Seventy five percent of the men reported normal interest in sexual activities and 87.5% reported normal erectile function; however, 4 of 16 reported a moderate loss of interest in sexual activities, and another 2 of 16 reported frequent loss of erectile function. All 4 men with decreased libido and both men with impaired erectile function had Hodgkin's disease. Fourteen (88%) of 16 patients had an elevated FSH level, 7 (47%) of 15 had elevated LH, and 6 (38%) of 16 had decreased testosterone levels. Decreased testosterone levels correlated with a moderate or total loss of libido (P = .008) and a diagnosis of Hodgkin's disease (P = .01). Thus, after transplantation, most men have abnormal levels of gonadotrophins. Decreased levels of testosterone and symptoms of sexual dysfunction correlated with a diagnosis of Hodgkin's disease and may be related to the induction and salvage therapy received prior to autoBMT. PMID- 11400951 TI - Tandem-cycle high-dose melphalan and cisplatin with peripheral blood progenitor cell support in patients with breast cancer and other malignancies. AB - We evaluated the feasibility of tandem-cycle high-dose chemotherapy (HDCT) with cisplatin, melphalan, and peripheral blood progenitor cells (PBPCs). Fifty patients with high-risk primary (n = 17) or stage IV breast cancer (n = 29) or other malignancies (n = 4) received 2 cycles of intravenous melphalan, 20 to 151.8 mg/m2, and cisplatin, 200 mg/m2, followed by granulocyte-macrophage colony stimulating factor (GM-CSF) or G-CSF. Starting at 40 mg/m2 of melphalan, patients also received PBPCs. Delayed platelet recovery defined the maximum tolerated dose (MTD) for melphalan at 101.2 mg/m2 per cycle. There were no treatment-related deaths. Cycle 2 was delivered at a median of 1.7 months after cycle 1; 72% of patients treated at the MTD received both cycles. Cycle 2 was omitted when patients refused it or had disease progression or toxicities, primarily prolonged thrombocytopenia. Complete response rates in stage IV breast cancer patients increased from 28% pre-HDCT to 55% after cycle 2. At a median follow-up of 4.6 years (range, 1.5-8.1 years), 11 of 29 patients with stage IV breast carcinoma were alive with 5-year projected progression-free and overall survival rates of 19% (95% confidence interval [CI], 7%-41%) and 39% (95% CI, 20%-62%), respectively. Five-year projected progression-free and overall survival rates for patients with stage IV breast cancer in complete response following HDCT versus all others were 35% (95% CI, 15%-70%) versus 0% (P = .01) and 61% (95% CI, 35% 91%) versus 10% (95% CI, 2%-60%) (P = .003; log-rank test), respectively. Estrogen-receptor positivity was predictive of reduced risk of progression (relative risk [RR], 0.25; 95% CI, 0.10-0.65; P = .003) and death (RR, 0.27; 95% CI, 0.10-0.72; P = .009) after adjusting for response status. Five-year projected relapse-free and overall survival rates were 71% (95% CI, 43%-96%) and 82% (95% CI, 56%-100%), respectively, for the 17 patients with high-risk primary breast cancer. Tandem-cycle high-dose melphalan and cisplatin with PBPCs is feasible. Preliminary data suggest significant activity in selected patients with stage IV responding breast carcinoma. PMID- 11400952 TI - High-dose therapy and autologous hematopoietic-cell transplantation for follicular lymphoma beyond first remission: the Stanford University experience. AB - A retrospective analysis was performed to investigate the outcome of high-dose therapy (HDT) and autologous hematopoietic cell transplantation in patients with follicular lymphomas beyond first remission. Ninety-two patients with primary induction failure or relapsed follicular low-grade lymphoma (FLGL), follicular large cell lymphoma (FLCL), and transformed follicular lymphoma (TFL) were treated with myeloablative therapy consisting of etoposide (60 mg/kg), cyclophosphamide (100 mg/kg), and either carmustine (BCNU;15 mg/kg) or fractionated total body irradiation (FTBI; 1200 cGy) followed by transplantation of purged autologous bone marrow or peripheral blood hematopoietic cells. For the 49 patients with relapsed FLGL, the median age was 49 years and the median interval from diagnosis to HDT was 30 months. The 4-year estimate of overall survival (OS) was 60% (95% confidence interval [CI], 45%-75%) and of disease-free survival (DFS) was 44% (95% CI, 29%-59%). Treatment with the FTBI-containing HDT regimen was associated with significantly longer DFS (P = .04) and OS (P = .04) in our multivariate analysis. OS was also significantly longer among those treated with 3 or fewer chemotherapy regimens. For the 26 FLCL patients, the median age was 51 years and in 31% the indication for HDT was primary induction failure. For FLCL patients, the 4-year estimate of OS was 58% (95% CI, 37%-79%) and of DFS was 51% (95% CI, 30%-72%). Among the 17 patients with TFL, 13 (76%) transformed at first relapse, and only 6 patients (35%) achieved complete remission with salvage therapy prior to HDT. For TFL patients, the 4-year estimate of OS was 50% (95% CI, 24%-76%) and of DFS 49% (95% CI, 20%-78%). There were 3 occurrences of myelodysplasia (1 after treatment with TBI, 2 after BCNU treatment), yielding an estimated incidence of 7% (95% CI, 0%-16%) at 56 months. This analysis shows that relapsed FLGL patients treated with 3 or fewer different chemotherapy regimens show inferior survival. The HDT regimen containing FTBI appears to be superior to the BCNU-based regimen for relapsed FLGL, although longer follow-up is needed to evaluate late effects. Lastly, patients with TFL or induction failure and relapsed FLCL can achieve survival outcome comparable to those observed with the indolent follicular lymphomas. PMID- 11400953 TI - Tobacco use among adults--Arizona, 1996 and 1999. AB - In 1994, Arizona passed the Tobacco Tax and Healthcare Act (Proposition 200) that increased the tax on cigarettes from $0.18 to $0.58, and allocated 23% of the resulting revenues to tobacco-control activities. Since 1995, Arizona has used the tobacco-control funds (approximately $30 million per year) to support the Arizona Department of Health Services (ADHS) Tobacco Education and Prevention Program (TEPP), a comprehensive program to prevent and reduce tobacco use. To track changes in tobacco use, the knowledge and opinions of Arizona residents about tobacco use, and the proportion of smokers advised to quit smoking by health-care providers, ADHS conducted the Arizona Adult Tobacco Survey (ATS) in 1996 and a follow-up survey in 1999. This report compares results of these two surveys, which indicate that prevalence of tobacco use among adults decreased, and the proportion of adults who were both asked about tobacco use and advised to quit by health-care providers and dentists increased. On the basis of these findings, if all states implemented comprehensive programs similar to those in Arizona, the national health objective for 2010 of reducing the adult smoking rate by half during this decade could be achieved. PMID- 11400954 TI - Protracted outbreaks of cryptosporidiosis associated with swimming pool use--Ohio and Nebraska, 2000. AB - Swimming is the second most popular exercise in the United States with approximately 400 million pool visits annually. During the summer of 2000, five outbreaks of cryptosporidiosis linked to swimming pools were reported to CDC. This report summarizes the investigations of two of these outbreaks involving approximately 1000 cases and provides recommendations to reduce the transmission of pool-related disease. PMID- 11400955 TI - Prevalence of parasites in fecal material from chlorinated swimming pools--United States, 1999. AB - As a result of the 1998 outbreak of infection with the chlorine-sensitive pathogen Escherichia coli O157:H7 at a waterpark in Georgia, many public health departments updated their guidelines for disinfecting pools following a fecal accident. Many of these guidelines recommended treating all fecal accidents as if they contained the highly chlorine-resistant parasite Cryptosporidium parvum, generally resulting in hyperchlorination and pool closures of up to a day. To determine whether fecal accidents commonly contained Cryptosporidium, the prevalence of this parasite and the moderately chlorine sensitive parasite Giardia intestinalis was assessed by asking swimming pool operators throughout the United States to collect formed stools from fecal accidents in their pools. This report summarizes the results of this study and provides recommendations for disinfecting pools following fecal accidents. PMID- 11400956 TI - Drowning--Louisiana, 1998. AB - Drowning is the third leading cause of death from unintentional injuries in Louisiana. In 1998, the fatality rate from drowning for Louisiana residents was 3.1 per 100,000 population, higher than the U.S. rate of 1.9 per 100,000, and more than twice the 2000 national target of 1.3 per 100,000 population. This report describes the demographics and risk factors associated with drownings in Louisiana in 1998. Findings indicate that alcohol or illicit drug use was found in approximately 60% of tested victims aged > or = 13 years and that none of the victims of boating-related drowning were correctly wearing a personal flotation device (PFD). Prevention efforts should focus on decreasing alcohol and illicit drug use and increasing the proper use of PFDs among boaters and others involved in water recreation. PMID- 11400957 TI - Responding to fecal accidents in disinfected swimming venues. AB - These recommendations are solely for management of fecal accidents in disinfected recreational water venues. The recommendations do not address use of other nonchlorine disinfectants because there is limited pathogen inactivation data for many of these compounds. Because improper handling of chlorinated disinfectants could cause injury, appropriate occupational safety and health requirements should be followed. PMID- 11400958 TI - Deferral of routine booster doses of tetanus and diphtheria toxoids for adolescents and adults. AB - A shortage of tetanus and diphtheria toxoids (Td) and tetanus toxoid (TT) in the United States has resulted because one of two manufacturers discontinued production of tetanus toxoid-containing products. Aventis Pasteur (Swiftwater, Pennsylvania) is the only major manufacturer of tetanus and Td in the United States. In response to the shortage, Aventis Pasteur has increased production of Td to meet national needs; however, because 11 months are required for vaccine production, the shortage is expected to last for the remainder of 2001. PMID- 11400959 TI - Compendium of Animal Rabies Prevention and Control, 2001. National Association of State Public Health Veterinarians, Inc. AB - The purpose of this compendium is to provide rabies information to veterinarians, public health officials, and others concerned with rabies prevention and control. These recommendations serve as the basis for animal rabies-control programs throughout the United States and facilitate standardization of procedures among jurisdictions, thereby contributing to an effective national rabies-control program. This document is reviewed annually and revised as necessary. Vaccination procedure recommendations are contained in Part I; all animal rabies vaccines licensed by the United States Department of Agriculture (USDA) and marketed in the United States are listed in Part II; Part III details the principles of rabies control. PMID- 11400960 TI - Trocar injuries in laparoscopic surgery. AB - BACKGROUND: Disposable trocars with safety shields are widely used for laparoscopic access. The aim of this study was to analyze risk factors associated with injuries resulting from their use as reported to the Food and Drug Administration. STUDY DESIGN: Manufacturers are required to report medical device related incidents to the Food and Drug Administration. We analyzed the 629 trocar injuries reported from 1993 through 1996. RESULTS: There were three types of injury: 408 injuries of major blood vessels, 182 other visceral injuries (mainly bowel injuries), and 30 abdominal wall hematomas. Of the 32 deaths, 26 (81%) resulted from vascular injuries and 6 (19%) resulted from bowel injuries. Eighty seven percent of deaths from vascular injuries involved the use of disposable trocars with safety shields and 9% involved disposable trocars with a direct viewing feature. The aorta (23%) and inferior vena cava (15%) were the vessels most commonly traumatized in the fatal vascular injuries. Ninety-one percent of bowel injuries involved trocars with safety shields and 7% involved direct-view trocars. The diagnosis of an enterotomy was delayed in 10% of cases, and the mortality rate in this group was 21%. In 41 cases (10%) the surgeon initially thought the trocar had malfunctioned, but in only 1 instance was malfunction subsequently found when the device was examined. The likelihood of injury was not related to any specific procedure or manufacturer. CONCLUSIONS: These data show that safety shields and direct-view trocars cannot prevent serious injuries. Retroperitoneal vascular injuries should be largely avoidable by following safe techniques. Bowel injuries often went unrecognized, in which case they were highly lethal. Device malfunction was rarely a cause of trocar injuries. PMID- 11400961 TI - Sentinel lymph node biopsy for breast cancer: impact of the number of sentinel nodes removed on the false-negative rate. AB - BACKGROUND: Numerous studies have demonstrated that sentinel lymph node (SLN) biopsy can accurately determine axillary nodal status for breast cancer, but unacceptably high false negative rates have also been reported. Attention has been focused on factors associated with improved accuracy. We have previously shown that injection of blue dye in combination with radioactive colloid reduces the false negative rate compared with injection of blue dye alone. We hypothesized that this may be from the increased ability to identify multiple sentinel nodes. The purpose of this analysis was to determine whether removal of multiple SLNs results in a lower false negative rate. STUDY DESIGN: The University of Louisville Breast Cancer Sentinel Lymph Node Study is a prospective multiinstitutional study. Patients with clinical stage T1-2, N0 breast cancer were eligible for enrollment. All patients underwent SLN biopsy using blue dye alone, radioactive colloid alone, or both agents in combination, followed by completion level I and II axillary dissection. RESULTS: A total of 1,436 patients were enrolled in the study from August 1997 to February 2000. SLNs were identified in 1,287 patients (90%), with an overall false negative rate of 8.3%. A single SLN was removed in 537 patients. Multiple SLNs were removed in 750 patients. The false negative rates were 14.3% and 4.3% for patients with a single sentinel node versus multiple sentinel nodes removed, respectively (p = 0.0004, chi-square). Logistic regression analysis revealed that use of blue dye injection alone was the only factor independently associated with identification of a single SLN (p<0.0001), and patient age, tumor size, tumor location, surgeon's previous experience, and type of operation were not significant. CONCLUSIONS: The ability to identify multiple sentinel nodes, when they exist, improves the diagnostic accuracy of SLN biopsy. Injection of radioactive colloid in combination with blue dye improves the ability to identify multiple sentinel nodes compared with the use of blue dye alone. PMID- 11400962 TI - The breast cancer patient with multiple sentinel nodes: when to stop? AB - BACKGROUND: During sentinel lymph node (SLN) biopsy for breast cancer, most authors report identifying a mean of 1 to 3 SLNs, but a range of 1 to 8 (or more) SLNs per patient. A significant minority of patients have 4 or more SLNs. Here we seek to determine the significance for the breast cancer patient of finding multiple SLNs, and whether there is an optimal threshold number of SLNs that should be removed. STUDY DESIGN: 1,561 patients who underwent successful SLN biopsy using blue dye and radioisotope in combination. Each SLN site was categorized prospectively by the operating surgeon as a dye success, an isotope success, or both. All SLNs containing counts at least four times greater than the postexcision axillary background were considered to be isotope successes. RESULTS: Fifteen percent of patients (241) had multiple (>3) SLNs. Ninety-eight percent of node-positive patients (440 of 449) were identified within the first three SLN sites examined. In 2% of all SLN positive patients (9 of 449) or 4% of patients with multiple SLN (9 of 241), a positive SLN was detected at site four or more. In eight patients the first positive SLN was found at sites four or more. Blue dye and isotope were equally effective in identifying metastases in patients with multiple SLNs. CONCLUSIONS: Fifteen percent of patients having SLN biopsy for breast cancer have multiple SLNs. Although 98% of positive SLNs were identified within the first three sites sampled, a small number of patients had their first positive SLN at sites 4 to 8. These data suggest that there is no absolute upper threshold for the number of SLNs that should be removed. Sampling a few additional SLNs probably adds little morbidity to the procedure, yet may significantly alter the treatment of some individuals. SLN biopsy should be continued until all blue and hot nodes are removed. PMID- 11400963 TI - The consequence of undertreating breast cancer in the elderly. AB - BACKGROUND: Recent studies have noted that a large fraction of elderly patients do not receive conventional treatment for breast cancer. The consequences of undertreatment of the elderly have not been adequately assessed. STUDY DESIGN: The senior author's database (PIT) was used to identify women undergoing potentially curative operations for breast cancer between 1978 and 1998. Risk factors, presentation, pathologic findings, treatment, and outcomes of 206 women aged over 70 years were compared with those of 920 younger patients. In addition, conventionally treated and "undertreated" elderly patients were identified, and their characteristics and outcomes were compared. RESULTS: Older patients' cancers were more often visible on mammography, usually as a mass; younger patients' mammograms were less frequently positive, presenting more often with calcifications (p = 0.002). Cancers of the elderly were better differentiated (p < 0.001) and more likely to be estrogen- and progesterone-receptor positive (p < 0.001; p = 0.007). Patients over 70 had fewer mastectomies (19% versus 33%; p < 0.001) and were also less likely to undergo axillary node dissection (71% versus 81%, p = 0.006), postoperative radiation (69% versus 92%, p < 0.001), and chemotherapy (18% versus 48%, p < 0.001). Fifty-seven percent of older patients were treated with tamoxifen compared with 36% of younger patients (p < 0.001). Elderly patients' rates of local and distant recurrence were comparable to those of younger patients after both mastectomy and breast conservation. Ninety-eight patients (54%) over 70 were undertreated by conventional criteria. Undertreated elderly patients were significantly older (78 versus 76 years, p = 0.003), were diagnosed with excisional biopsy more often (69% versus 57%, p = 0.069) and with fine-needle aspiration less frequently (22% versus 38%, p = 0.069), and were more likely to have breast conservation (90% versus 73%, p = 0.004). Local and distant disease-free survival rates of both groups were comparable. Tamoxifen treatment significantly reduced the chance of developing distant metastasis in node negative elderly patients with invasive tumors (p = 0.028). Omission of chemotherapy had no impact on disease control in the elderly. Axillary node status and estrogen-receptor status were significantly related to local disease free survival, and axillary node status was significantly related to distant disease-free survival in multivariate analysis in the elderly. CONCLUSIONS: Elderly breast cancer patients are frequently treated with breast conservation, omitting axillary dissection, radiation therapy, and chemotherapy. Despite undertreatment by conventional criteria, the rates of local recurrence and distant metastasis are not increased in comparison with conventionally treated elderly patients. Tamoxifen should be administered to elderly breast cancer patients with invasive tumors because it significantly improves distant control. PMID- 11400964 TI - Prognostic significance of CyclinD1 and E-Cadherin in patients with esophageal squamous cell carcinoma: multiinstitutional retrospective analysis. Research Committee on Malignancy of Esophageal Cancer, Japanese Society for Esophageal Diseases. AB - BACKGROUND: Although many molecular biologic molecules have been analyzed for their prognostic influence on patients with esophageal cancer, previous studies have not been able to raise statistically significant prognostic factors. STUDY DESIGN: Immunohistochemical analysis of CyclinD1 expression and E-Cadherin expression was performed retrospectively in 416 esophageal squamous cell cancer patients who underwent curative resection of esophageal cancer at 10 major surgical departments in Japan, where more than 30 esophagectomies are performed in a year. The prognostic impact of these molecules and their relationship to clinicopathologic data of the patients were evaluated. RESULTS: Univariate analysis revealed that pN (pTNM), pT (pTNM), CyclinD1 expression, and E-Cadherin expression were significant prognostic factors, and multivariate analysis revealed that pN (risk ratio (RR) 2.19), pT (RR 3.35), CyclinD1 (RR 1.42), and E Cadherin (RR 0.71) were significant prognostic factors. Combination analysis of these genes revealed that E-Cadherin-preserved and CyclinD1-negative patients had the best prognosis; E-Cadherin-reduced and CyclinD1-positive patients had the worst prognosis. CONCLUSIONS: Increased CyclinD1 expression and reduced E Cadherin expression were significant prognostic factors in patients with esophageal squamous cell carcinoma. PMID- 11400965 TI - Emergency surgery for obstructing colorectal cancers: a comparison between right sided and left-sided lesions. AB - BACKGROUND: Fifteen to twenty percent of patients with primary colorectal cancers present with intestinal obstruction. Traditionally, different approaches have been used in the management of right-sided and left-sided colonic obstruction. Recently, single-stage resection with primary anastomosis in left colonic obstruction has been shown to have good results. The objective of this study was to compare the operative results of patients who had emergency operations for right-sided and left-sided obstructions from primary colorectal cancers. STUDY DESIGN: This is a retrospective study including 243 patients who underwent emergency operations for obstructing colorectal cancers from 1989 to 1997. Primary resection of the tumor-bearing segment followed by primary anastomosis was attempted when the conditions were feasible. The operative results of patients with right-sided tumors were compared with those of patients with left sided tumors. RESULTS: One hundred seven patients had obstruction at or proximal to the splenic flexure (right-sided lesions), and 136 had lesions distal to the splenic flexure (left-sided lesions). The primary resection rate was 91.8%. Of the 223 patients with primary resection, primary anastomosis was possible in 197 patients. Among the 101 primary anastomoses in patients with left-sided obstruction, segmental resection with on-table lavage was performed in 75 patients and subtotal colectomy was performed in 26. The overall operative mortality rate was 9.4%, although that of the patients with primary resection and anastomosis was 8.1%. The anastomotic leakage rate for those with primary resection and anastomosis was 6.1%. There were no differences in the mortality or leakage rates between patients with right-sided and left-sided lesions (mortality: 7.3% versus 8.9%, p = 0.79; leakage: 5.2% versus 6.9%, p = 0.77). Colocolonic anastomosis did not show a significant difference in leakage rate when compared with ileocolonic anastomosis (6.1% versus 6.0%, p = 1.0). CONCLUSIONS: This study showed that primary resection and anastomosis for left sided malignant obstruction, either by segmental resection with on-table lavage or subtotal colectomy, was not more hazardous than primary anastomosis for right sided obstruction. The single-stage procedure should be the objective for the treatment of patients with obstructing colorectal cancers, except when patients are hemodynamically unstable during surgery or when the condition of the bowel is not optimal for primary anastomosis. PMID- 11400966 TI - The effect of preoperative biliary drainage on postoperative complications after pancreaticoduodenectomy. AB - BACKGROUND: The benefit of preoperative biliary drainage in jaundiced patients undergoing pancreaticoduodenectomy for a suspected malignancy of the periampullary region is still under debate. This study evaluated preoperative biliary drainage in relation to postoperative outcomes. STUDY DESIGN: At the Academic Medical Center, Amsterdam, the Netherlands, a cohort of 311 patients undergoing pancreaticoduodenectomy from June 1992 up to and including December 1999 was studied. Of this cohort 21 patients with external or surgical biliary drainage were excluded and 232 patients who had received preoperative internal biliary drainage were divided into three groups corresponding with severity of jaundice according to preoperative plasma bilirubin levels: < 40 microM (n = 177), 40 to 100 microM (n = 32), and > 100 microM (n = 23) were designated as groups 1, 2, and 3, respectively. These groups were compared with patients who underwent immediate surgery (n = 58) without preoperative drainage. RESULTS: The median number of stent (re)placements was 2 (range 1 to 6) with a median drainage duration of 41 days (range 2 to 182 days) and a stent dysfunction rate of 33%. Although patients in group 1 were better drained than patients in groups 2 and 3 (median reduction of bilirubin levels 82%, 57%, and 37%, respectively, p < 0.01), there was no difference in overall morbidity among the drained groups (50%, 50%, and 52%, respectively). There was no significant difference in overall morbidity between patients with and without preoperative biliary drainage (50% and 55%, respectively). CONCLUSIONS: Preoperative biliary drainage did not influence the incidence of postoperative complications, and although it can be performed safely in jaundiced patients it should not be used routinely. PMID- 11400967 TI - Oral examinations and grading sessions promote faculty and resident enthusiasm for student evaluation and teaching. PMID- 11400968 TI - What's new in general thoracic surgery. PMID- 11400969 TI - What's new in otolaryngology. PMID- 11400970 TI - The Lynn Sage Memorial Lecture: new developments in breast cancer management. PMID- 11400971 TI - The surgical suite meets the new health economy. PMID- 11400972 TI - Fundamentals of contemporary surgical oncology: biologic principles and the threshold concept govern treatment and outcomes. PMID- 11400973 TI - Correlation of functional MRI with intraoperative cortical mapping in patient with cerebral arteriovenous malformation. PMID- 11400974 TI - Gangliocytic paraganglionoma of the duodenum. PMID- 11400975 TI - Gastroesophagostomy after proximal gastrectomy using a double stapling technique. PMID- 11400976 TI - Duct-to-duct biliary reconstruction in right lobe adult living donor liver transplantation. PMID- 11400977 TI - Solitary hepatic cysts. PMID- 11400978 TI - Frequency of prophylactic contralateral mastectomy among breast cancer patients. PMID- 11400979 TI - Scientists find new tumor suppressor gene involved in breast, prostate, and other cancers. PMID- 11400980 TI - Scientists repair damage from heart attack using adult bone marrow stem cells in mice. PMID- 11400981 TI - FSA sets out its five-year plan. PMID- 11400982 TI - FMD: CVO expresses concern about Yorkshire outbreaks. PMID- 11400983 TI - Foot-and-mouth disease: lessons for the future. PMID- 11400984 TI - Use of a reverse-transcriptase polymerase chain reaction for monitoring the shedding of feline coronavirus by healthy cats. AB - The pattern of shedding of feline coronavirus (FCoV) was established in 155 naturally infected pet cats from 29 households over periods of up to five years. Viral RNA was detected in faeces by reverse-transcriptase PCR (RT-PCR), and plasma antiviral antibodies by immunofluorescence. The cats rarely shed FCoV in their saliva. Three patterns of FCoV shedding were observed. Eighteen of the cats shed virus continuously, so were persistent, and possibly lifelong, carriers; none of them developed feline infectious peritonitis. Fifty-six cats ceased shedding virus, although they were susceptible to reinfection, and 44 shed intermittently or were being continuously reinfected. Four of the cats were resistant to infection. Seventy-three per cent of the virus shedding episodes lasted up to three months and 95 per cent up to nine months. There was a correlation between shedding and antibody titre but the cats could remain seropositive for some time after they had ceased shedding virus. One-off testing for FCoV by RT-PCR is inappropriate. Identification of longterm carriers requires that a positive result be obtained by RT-PCR on faecal samples for at least eight consecutive months. A cat should be shown to be negative over five months, or to have become seronegative, to ensure that it has ceased shedding virus. PMID- 11400985 TI - Aetiology of reduced milk ejection in cows after transport and the use of a long acting analogue of oxytocin for prophylaxis. AB - Milk flow was recorded in 21 cows for three days after they were admitted to a large animal hospital. When the spontaneous flow of milk had stopped, a physiological dose (1 iu) of oxytocin was administered intravenously. Five of the cows were, in addition, treated with 0.35 mg of a long-acting analogue of oxytocin (carbetocin) one hour before the first milking after they were admitted. In the 16 cows not treated with carbetocin, only about 30 per cent of the total milk yield was released spontaneously on the first day, and the injection of 1 iu of oxytocin released approximately another 60 per cent of the total milk yield. On the second day, the proportion of the total milk yield released spontaneously increased and the fraction released after the injection of 1 iu oxytocin decreased. In contrast, the five cows treated with carbetocin released on average 94 per cent of the total milk yield spontaneously during the first milking. PMID- 11400986 TI - Serological evidence of Brucella species infection in odontocetes from the south Pacific and the Mediterranean. AB - Sera from 58 odontocetes taken in fisheries off Peru in 1993 to 1995 and from 24 cetaceans stranded along the Spanish coast of the Mediterranean in 1997 to 1999 were tested for the presence of Brucella species antibodies in competitive and indirect ELISAS (cELISA and iELISA). Among the animals from Peru, 21 of 27 (77.8 per cent) Lagenorhynchus obscurus, three of six Delphinus capensis, one of two inshore and two of three offshore Tursiops truncatus and five of 20 (25 per cent) Phocoena spinipinnis were positive in the cELISA. Brucella species antibodies were also observed in two of 16 (12.5 per cent) Stenella coeruleoalba and in one of two Ttruncatus from the Mediterranean. These data provide the first evidence for the presence of cetacean brucellae in the south Pacific Ocean and the Mediterranean Sea. PMID- 11400987 TI - Treatment of canine atopic dermatitis with cyclosporine: a pilot clinical study. PMID- 11400988 TI - Report on 31 caseous lymphadenitis infected sheep farms in England and Wales. PMID- 11400989 TI - Seroprevalence of Ehrlichia canis antibodies among dogs in Turkey. PMID- 11400990 TI - Outbreaks of contagious agalactia in small ruminants in the Republic of Macedonia. PMID- 11400991 TI - Implementaton of 'the TSE directive'. PMID- 11400992 TI - FMD control strategies. PMID- 11400993 TI - FMD control strategies. PMID- 11400994 TI - FMD control strategies. PMID- 11400995 TI - Learning from FMD. PMID- 11400996 TI - Burring elodont cheek teeth in small herbivores. PMID- 11400997 TI - Veterinary public health teaching. PMID- 11400998 TI - Effect of oral nicotine dosing forms on cigarette withdrawal symptoms and craving: a systematic review. AB - RATIONALE: Oral nicotine dosing forms such as nicotine gum have been found to be effective in helping smokers to stop. Some, but not all studies have also found that they reduce the severity of withdrawal discomfort. With new oral nicotine products being developed and use of existing products widening, it is important to determine the strength of evidence that these forms of nicotine replacement reduce overall withdrawal discomfort and individual withdrawal symptoms including craving. OBJECTIVES: To assess the strength of evidence that oral nicotine reduces the severity of overall withdrawal discomfort and individual withdrawal symptoms, including craving. METHODS: All published studies reporting effects of nicotine gum, inhaler, lozenge and sublingual tablet on recognised withdrawal symptoms were scanned and those that met a set of quality criteria were included in the review. The key characteristics of these studies were summarised and their findings tabulated. RESULTS: Of 27 studies that reported effects of oral nicotine products on at least one withdrawal symptom, 12 met the quality criteria (eight for nicotine gum, three for inhaler, one for microtab, none for 1 mg lozenge). Because of limitations on the reporting of the studies it was not possible to carry out meta-analyses and good data were available only for the first week of abstinence. Six out of seven studies reporting it found an effect on total withdrawal discomfort, nine out of nine found an effect on irritability, three out of four found an effect on anxiety and the only study that looked at it found an effect on depressed mood. Seven of 11 studies that looked at it found an effect on craving, but only three out of seven of the studies of nicotine gum. For other withdrawal symptoms the findings were more mixed. CONCLUSIONS: We conclude that the strength of evidence that oral nicotine forms reduce total withdrawal discomfort, irritability and anxiety is high. There is some evidence for an effect on depressed mood and craving although in the latter case the evidence is less good for gum than other forms. Future studies of nicotine effects on withdrawal should meet minimum quality standards for design and reporting to enable results to be combined and compared across studies. PMID- 11400999 TI - Tryptophan depletion in SSRI-recovered depressed outpatients. AB - RATIONALE: Recently, a number of studies have challenged the finding that acute tryptophan depletion (TD) increases depressive symptoms in medicated, formerly depressed patients. The present study examined the effects of acute nutritional TD on remitted depressed patients currently treated with selective serotonin reuptake inhibitors. In an attempt to clarify conflicting earlier findings, the effects of a number of clinical variables on outcome were also investigated. METHODS: Ten patients underwent TD in a double-blind, controlled, balanced crossover fashion. The control session followed the procedure of Krahn et al. (1996 Neuropsychopharmacology 15:325-328). Sessions were 5-8 days apart. RESULTS: TD was significantly related to increased scores on clinician-rated depression and anxiety scales, and on self-rated depression, anxiety, and somatic symptoms. The control challenge had no effect, despite the fact that the reductions in plasma tryptophan during the control session were unexpectedly high. Some evidence was found for a threshold in the relationship between reduction of plasma tryptophan and mood response. CONCLUSIONS: The mood effect of TD in medicated, formerly depressed patients was confirmed. A threshold may exist for mood effects following TD, implying that recent negative findings may have been caused by insufficient depletion. No other predicting or mediating factors were identified, although the variable "history of response pattern to medication" deserves further study. PMID- 11401000 TI - Intramuscular (IM) ziprasidone 20 mg is effective in reducing acute agitation associated with psychosis: a double-blind, randomized trial. AB - RATIONALE: Intramuscular (IM) conventional antipsychotics and/or benzodiazepines are effective in the short-term treatment of acutely agitated psychotic patients but may be associated with adverse effects. A short-acting IM formulation of the novel antipsychotic, ziprasidone, which may offer advantages over conventional agents, has been developed. OBJECTIVE: To compare ziprasidone IM 2 mg (n=38) and 20 mg (n=41) in the acute control and short-term management of agitated psychotic patients. METHODS: A prospective, randomized, double-blind, 24-h study assessed efficacy using the Behavioral Activity Rating Scale (BARS) and the PANSS. The BARS is a validated rating scale for the assessment of treatment response in acute agitation associated with psychosis. Following the initial dose, three more doses could be given 4 h apart if needed during the 24-h period. RESULTS: The mean BARS score had decreased 15 min after the first 20 mg IM dose and was statistically significantly lower than the 2 mg group at 30 min post-dose. The improvement with the 20 mg dose increased until 2 h, and was maintained until at least 4 h post-dose (P<0.001). Two hours after the first injection, almost all of the patients receiving ziprasidone 20 mg were BARS responders compared with just one-third of those receiving 2 mg ziprasidone (P<0.001). The calming effect of ziprasidone was also evident by the significant reduction in PANSS agitation items (P<0.05) and CGI-severity at 4 h (P=0.008). Both ziprasidone doses were very well tolerated. Ziprasidone IM 20 mg was not associated with EPS, dystonia, akathisia, respiratory depression or with excessive sedation. CONCLUSIONS: Ziprasidone IM 20 mg substantially and significantly reduced the symptoms of acute agitation in patients with psychotic disorders. Ziprasidone 20 mg IM was very well tolerated and produced no dystonia or akathisia. PMID- 11401001 TI - Clozapine enhances breakpoint in common marmosets responding on a progressive ratio schedule. AB - RATIONALE: Motivational effects of psychotropic drugs may contribute to their therapeutic profile and progressive ratio (PR) schedules provide a method of measuring these effects in animals. OBJECTIVE: Determine effects of selected antipsychotic, psychotomimetic, anxiolytic and antidepressant drugs on PR performance in common marmosets. METHOD: Marmosets were trained to lever press for banana milkshake reinforcement using a PR schedule, in which the number of lever presses to achieve successive reinforcements increased by one until responding ceased (breakpoint). RESULTS: Clozapine administered intramuscularly (0.01-2 mg/kg IM; 30 min pretreatment time (ptt) or by oral gavage (0.1-4 mg/kg PO; 60 min ptt) significantly increased the breakpoint. Independent tests of fluid consumption failed to show enhanced fluid intake after clozapine pretreatment, suggesting this effect was not due to drug induced polydipsia. Neither haloperidol (0.005-0.1 mg/kg PO; 60 min ptt) nor risperidone (0.0025-0.05 mg/kg PO; 60 min ptt) altered breakpoint. Olanzapine (0.01-1 mg/kg PO; 60 min ptt) significantly enhanced the breakpoint at 0.05 mg/kg PO, but the effect was not robust. Amphetamine (0.2-2 mg/kg SC; 30 min ptt) significantly reduced the breakpoint at 2 mg/kg and fluoxetine (0.1-1 mg/kg PO; 60 min ptt) was without effect. Diazepam significantly increased the breakpoint at 0.5 mg/kg PO. Drug induced polydipsia might play a role in this response as independent tests showed increased fluid consumption following diazepam. CONCLUSIONS: These results demonstrate that, unlike other antipsychotics, clozapine over a wide dose range increased the motivational state of marmosets to respond for banana milkshake. This motivational aspect of clozapine's actions may contribute to its unique clinical profile and the PR procedure may provide a method for detecting novel antipsychotics with a clozapine-like profile. PMID- 11401002 TI - Pharmacological evidence of muscarinic-cholinergic sensitization following chronic stress. AB - RATIONALE: Although much evidence supports a major role of brain cholinergic transmission in memory consolidation processes, little is known about cholinergic functioning under environmental pressure. OBJECTIVES: The present experiments were aimed at investigating possible functional adaptation of muscarinic receptors promoted by a chronic stressful procedure in an inbred strain of mice highly susceptible to stress. METHODS: We tested the effects of post-trial administration of a cholinergic agonist and a muscarinic antagonist on the retention of a passive avoidance task in control animals and compared these effects with those observed following food restriction. RESULTS: Food restriction enhanced the facilitatory effects of oxotremorine and reduced the impairing effects of atropine on memory consolidation. CONCLUSION: Our results support the view that chronic sensitization of muscarinic receptors occurs following chronic stress. PMID- 11401003 TI - Pharmacokinetics of reboxetine in healthy volunteers with different ethnic descents. AB - RATIONALE: Ethnicity can affect the pharmacokinetics and pharmacodynamics of psychopharmacologic drugs. OBJECTIVES: Reboxetine disposition differences among Asians, blacks, and Caucasians were examined. METHODS: Healthy subjects (12 Asians, 12 blacks, 12 Caucasians) received a single oral dose of one 4-mg reboxetine tablet in an open label, parallel study design. Plasma concentrations of reboxetine enantiomers [R,R(-) reboxetine and predominantly active S,S(+) reboxetine] were quantified using HPLC-MS-MS. Plasma unbound fractions of reboxetine enantiomers were evaluated by equilibrium dialysis. Ethnic group effects on pharmacokinetic parameters were assessed by ANOVA. RESULTS: Mean S,S(+) reboxetine CLPO for blacks was significantly greater, compared to Asians and Caucasians (154+/-82 ml/min, 101+/-19 ml/min and 101+/-18 ml/min, respectively). Mean S,S(+) reboxetine free fractions (fu) were significantly greater for Asians and blacks, compared to Caucasians (3.04+/-1.28%, 2.89+/ 0.69%, and 1.99+/-0.58%, respectively). S,S(+) Reboxetine unbound clearance (CLu) was significantly less for Asians, compared to blacks and Caucasians (3742+/-1468 ml/min, 5187+/-2027 ml/min, and 5294+/-1163 ml/min, respectively). S,S(+) Reboxetine mean unbound AUC (AUCu) in these groups were 20.2+/-7.1 ng.h/ml, 14.6+/-5.1 ng.h/ml, and 13.2+/-3.2 ng.h/ml, respectively. AUCu was significantly greater for Asians. CLu and AUCu did not differ significantly between blacks and Caucasians. Ethnic effects of R,R(-) reboxetine were similar to those observed for S,S(+) reboxetine. CONCLUSIONS: The AUCu difference between Asian and black and Caucasian subjects was modest. Tolerability differences among groups were not observed. No dosage adjustment is necessary for Asians or blacks. PMID- 11401004 TI - Conditioned antinociception and freezing using electrical stimulation of the dorsal periaqueductal gray or inferior colliculus as unconditioned stimulus are differentially regulated by 5-HT2A receptors in rats. AB - RATIONALE: Electrical stimulation of the dorsal periaqueductal gray matter (dPAG) and the inferior colliculus (IC) has been used as an aversive unconditioned stimulus. However, studies on the behavioral, sensorial and autonomic components of the conditioned fear elaborated in the midbrain tectum are lacking. OBJECTIVES: This study was undertaken to investigate the nature of the aversiveness of stimulation of the dPAG and IC as well as the modulation by 5-HT mechanisms of the fear conditioned responses to these stimulations. METHODS: Animals chronically implanted with an electrode glued to a guide cannula into the dPAG or the IC were submitted to one, two or three sessions of conditioning. Each session consisted of ten pairings of the light in a distinctive chamber (CS) with the electrical stimulation of one of these regions at the escape threshold determined previously. Control groups were submitted to the same procedure, except for the conditioning sessions in which the conditioned stimuli were presented alone in one case and performed in a different context in the other. On the next day, each animal was exposed only to the CS (testing) and the duration of freezing, number of rearings, grooming, bouts of micturition and fecal boli were recorded for 5 min. Before and after the testing session, the animals were submitted to the tail-flick test. RESULTS: The data showed that the conditioning with electrical stimulation of the dPAG and the IC caused significant increases in the time of freezing, defecation and micturition, and significant reductions in the number of rearings and grooming. On the other hand, only the conditioning with electrical stimulation of the dPAG produced significant conditioned antinociception. Microinjections of methysergide, a non-specific antagonist of 5 HT receptors, or ketanserin, an antagonist of 5-HT2A receptors, into the dPAG before testing significantly inhibited the antinociception without affecting any of the behavioral or autonomic conditioned responses. CONCLUSIONS: 1) Conditioned freezing may be produced using the electrical stimulation of the dPAG or IC as unconditioned stimuli, 2) only the pairing of CS plus dPAG but not with IC stimulation, produces significant conditioned antinociception, 3) blockade of 5 HT2A receptors inhibits conditioned antinociception but not the conditioned defensive behavior using the electrical stimulation of the dPAG as unconditioned stimulus. PMID- 11401005 TI - Threshold doses for nicotine discrimination in smokers and non-smokers. AB - RATIONALE: Tobacco use during initial experimentation often involves modest nicotine exposure, escalating to larger doses and more frequent exposure with the onset of tobacco dependence. Threshold doses for nicotine discrimination therefore may differ between naive and experienced tobacco users. OBJECTIVES: We determined the lowest (threshold) dose of nasal spray nicotine that smokers and non-smokers could reliably discriminate from placebo spray. METHODS: Male and female smokers (n=18) and non-smokers (n=17) were initially trained to discriminate 20 microg/kg from placebo before proceeding to threshold determination sessions, which involved discrimination of progressively lower doses below 20 microg/kg ("descending order" subgroup) or higher doses above 1 microg/kg ("ascending order" subgroup). Threshold was determined by the lowest dose reliably discriminated from placebo (correct on > or =80% of testing trials) and by failure to discriminate the next lowest dose. RESULTS: Threshold doses for nicotine discrimination were low and not different between smokers and non smokers (median thresholds of 3 versus 2 microg/kg and approximate blood levels of 2.6 versus 1.6 ng/ml, respectively). Thresholds were similar between descending and ascending order subgroups. Several subjective responses differentiated threshold dose from the dose just below threshold, particularly in non-smokers. CONCLUSIONS: Threshold doses for nasal spray nicotine discrimination in humans are low, well below the typical nicotine delivery of most cigarette brands, and may not change after long-term smoking exposure. PMID- 11401006 TI - Bupropion SR worsens mood during marijuana withdrawal in humans. AB - RATIONALE: Symptoms of withdrawal after daily marijuana smoking include increased ratings of irritability and depression. Similar mood symptoms are reported by cigarette smokers during nicotine abstinence. OBJECTIVE: Given the successful use of sustained-release bupropion in treating nicotine dependence, this study investigated how maintenance on bupropion influenced symptoms of marijuana withdrawal compared to maintenance on placebo. METHODS: Marijuana smokers (n=10) were maintained outpatient on active (300 mg/day) or placebo (0 mg/day) bupropion for 11 days, and were then maintained inpatient on the same bupropion dose for 17 days. For the first 4 inpatient days, participants smoked active marijuana [2.8% delta9-tetrahydrocannabinol (THC)] 5 times/day. For the remaining inpatient days, participants smoked placebo marijuana (0.0% THC) 5 times/day. Participants were then maintained outpatient on the alternate dose of bupropion for 11 days, followed by a second inpatient residential stay, paralleling the first. Medication administration was double-blind. Mood, psychomotor task performance, food intake, and sleep were measured daily during each inpatient phase. The order of active and placebo bupropion maintenance was counterbalanced between groups. RESULTS: Bupropion had few behavioral effects when participants smoked active marijuana. During placebo marijuana smoking, i.e., active marijuana withdrawal, ratings of irritability, restlessness, depression, and trouble sleeping were increased by bupropion compared to placebo maintenance. CONCLUSIONS: These data suggest that bupropion does not show promise as a potential treatment medication for marijuana dependence. PMID- 11401007 TI - Differential interaction of GBR 12909, a dopamine uptake inhibitor, with cocaine and methamphetamine in rats discriminating cocaine. AB - RATIONALE: Inhibitors of neuronal dopamine uptake, such as GBR 12909, decrease IV cocaine self-administration by laboratory animals and have been proposed as potential therapeutic agents for abuse of psychomotor stimulant drugs. OBJECTIVES: This study was performed to determine how GBR 12909 alters the discriminative stimulus effects of methamphetamine and cocaine. METHODS: Rats were trained to discriminate between IP injections of 10 mg/kg cocaine and saline and were tested for stimulus generalization to cocaine, GBR 12909, and methamphetamine. Based upon the ED50 of the individual drugs, combinations of GBR 12909 and either cocaine or methamphetamine were tested that comprised a) 1 part GBR 12909 and 2 parts cocaine or methamphetamine, or b) 2 parts GBR 12909 and 1 part cocaine or methamphetamine. RESULTS: GBR 12909 and cocaine were equipotent and 30-fold less potent than methamphetamine in producing cocaine-like discriminative effects. GBR 12909 and cocaine produced cocaine-like discriminative effects synergistically in the ratio of 1 part GBR 12909:2 parts cocaine (0.16+0.32 to 1.92+ 3.87 mg/kg) and nearly synergistically in the ratio of 2 parts GBR 12909:1 part cocaine (0.32+0.16 to 3.92+ 1.91 mg/kg). GBR 12909 and methamphetamine (0.32+0.02 to 3.20+0.22 mg/kg or 0.65+0.01 to 6.53+0.1 mg/kg) were simply additive in both sets of fixed-ratio dose combinations. CONCLUSIONS: The synergy of GBR 12909 and cocaine and the additivity of GBR 12909 and methamphetamine run counter to the presumed mechanisms of action of these drugs at dopamine nerve terminals, which might have implications for the use of GBR 12909 in the treatment of addiction to cocaine or amphetamines. PMID- 11401008 TI - Distinct temporal pattern of the effects of the combined serotonin-reuptake inhibitor and 5-HT1A agonist EMD 68843 on the sleep EEG in healthy men. AB - RATIONALE: EMD 68843 (EMD) has properties of a serotonin (5-HT)-reuptake inhibitor and a partial 5-HT1A agonist in one molecule in order to combine antidepressive and anxiolytic properties. OBJECTIVE: We investigated the effects of EMD on the sleep EEG in order to characterize how the complex interaction between these two properties influences the sleep EEG. METHODS: We performed a randomized crossover study in ten young normal male controls (20-30 years), receiving a single dose of 20 mg EMD or placebo orally at 2100 hours. Sleep EEG was recorded from 2300 to 0700 hours. RESULTS: After EMD, rapid eye movement (REM) sleep was nearly totally abolished. In the course of the night other effects on the sleep EEG occurred in distinct intervals. Slow wave sleep and EEG delta power increased in the first and third one-third of the night, whereas wakefulness was enhanced in the second and third one-third of the night. CONCLUSION: The sleep EEG effects of EMD fit with its pharmacological profile, which might lead to adaptive changes suggested to characterize an antidepressive substance. PMID- 11401009 TI - Quantifying drug-related 5-HT1A receptor occupancy with. AB - RATIONALE: There is an increased interest in measuring the interaction of new or established drugs with their targets, in order to gain a better understanding of their mechanisms of action. PET can provide this information if an appropriate radioligand is available. [18F]MPPF (4-(2'-methoxyphenyl)-1-[2'-(N-2"-pyridinyl) p-[18F]fluorobenzamido]ethylpiperazine) is a selective radioligand for serotonin 5-HT1A receptors. We have established that the binding potential (BP=Bmax/KD) of [18F]MPPF for cerebral 5-HT1A receptors can be assessed in human brain without arterial sampling. OBJECTIVES: The aim of this study was to assess if 5-HT1A receptor occupancy can be measured through calculation of a drug-related decrease in BP with [18F]MPPF and PET. METHODS: Six volunteers were scanned twice using a Siemens Exact HR+ camera following injection of 70+/-18 MBq [18F]MPPF (baseline and medicated conditions). Before the second scan, volunteers orally received either 3x10 mg pindolol at T=-15.5 h, T=-6.5 h, and T=-1.5 h (n=3) or 10 mg buspirone in a single dose at T=-1.5 h (n=3). Binding potentials were calculated using the simplified reference tissue model with the cerebellum as reference. RESULTS: Administration of 30 mg pindolol led to a significant reduction in [18F]MPPF binding potential of 42+/-17%. In contrast, no significant reduction of [18F]MPPF binding potential was observed following administration of buspirone (5+/-17%). CONCLUSIONS: These results show that [18F]MPPF can be used for measurement of drug-related 5-HT1A receptor occupancy and may be of particular interest in determining the 5-HT1A receptor interaction of new or established drugs in phase 1 and early phase 2 drug trials. Apparently, the 5-HT1A partial agonist buspirone is already clinically effective at low levels of 5-HT1A receptor occupancy. PMID- 11401010 TI - Relationship between variations in estradiol and progesterone levels across the menstrual cycle and human performance. AB - RATIONALE: Studies about whether or not the cognitive performance of women is influenced by changes in levels of sex steroid hormones across the menstrual cycle have produced ambiguous results. OBJECTIVES: This study tested whether flight simulator performance differs significantly between the menstrual and the luteal phase of the menstrual cycle. METHODS: In a within-subjects design, 24 female pilots were tested twice during their menstrual cycle: once during the menstrual and once during the luteal phase. On both test days they performed a 75 min simulator flight in a Frasca 141, a popular pilot training device. RESULTS: Despite highly significant differences in estradiol (E2) as well as progesterone (P) levels on the 2 test days, and despite excluding subjects with anovulatory cycles from the analyses, there were no significant differences in overall flight performance between the menstrual and luteal phases. We found no significant correlations between E2 or P levels and flight performance. CONCLUSIONS: We found no evidence that the tested menstrual cycle phases and their associated E2 and P levels significantly influence flight simulator performance. We consider these negative findings based on 24 subjects meaningful because previous studies on the influence of menstrual cycle on cognitive performance have not involved complex "real world" tasks such as piloting an aircraft and they obtained inconsistent results. PMID- 11401011 TI - Lorazepam and diazepam impair true, but not false, recognition in healthy volunteers. AB - RATIONALE: The deleterious effects of benzodiazepine on memory are well documented. However, their effects on false memories are unknown. OBJECTIVE: The aim of this study was to investigate the effects of lorazepam and diazepam on false memories and related states of awareness in healthy volunteers. METHODS: The Deese/Roediger-McDermott procedure was used in 36 healthy volunteers randomly assigned to one of three parallel groups (placebo, diazepam 0.3 mg/kg, lorazepam 0.038 mg/kg). Subjects studied lists of words semantically related to a non presented theme word (critical lure). On a recognition memory task with both previously presented words and non presented critical lures, they were asked to give Remember, Know or Guess responses to items that were recognized on the basis of conscious recollection, familiarity, or guessing, respectively. RESULTS: The proportions of studied words correctly recognized and the proportions of Remember responses associated with true recognition were lower in the benzodiazepine groups than in the placebo group. In contrast, benzodiazepines did not significantly influence the proportions of critical lures falsely recognized or the proportions of Remember responses associated with false recognition. CONCLUSION: These results indicate that diazepam and lorazepam impair conscious recollection associated with true, but not false, memories. PMID- 11401012 TI - Cholinergic modulation of covert attention in the rat. AB - RATIONALE: Nicotine is known to facilitate attentional processing, but its role in processing spatial and non-spatial cues is not well established in rodents. OBJECTIVE: These experiments tested the hypothesis that nicotine facilitates the orienting of attention in space but has no effect on non-spatial cues and that the benefits are blocked by the nicotinic antagonist mecamylamine. METHODS: Eight male rats were trained to insert their noses in an opening, which triggered the presentation of cue and target lights in a modified covert orienting task. Four types of trials were presented: valid cues (cue and target lights on the same side of the nose hole), invalid cues (cue and targets on opposite sides), double cues (both cue lights illuminated, target on either side), and no cue (cue lights omitted, targets on either side). The reaction time required to withdraw the nose from the fixation hole (RT) and the time for the rat to move to the feeder (MT) were measured. RESULTS: Nicotine decreased all RTs in a dose-dependent manner but significantly lowered the invalid cue RTs and the validity effect (invalid-valid cue RT). Mecamylamine slowed RTs in a dose-dependent fashion and reduced the validity effect by significantly slowing the valid cue RTs. With mixtures of a fixed strength of nicotine and an increasing dose of mecamylamine, RTs showed nicotine-like effects at low doses and mecamylamine-like effects at high doses. Neither of these drugs had a major effect on non-orienting trials (double and no cue RTs), and the alerting effect (no cue RTs-double cue RTs). CONCLUSIONS: Taken together with recent work in humans and non-human primates, these results suggest that the nicotinic cholinergic modulation of visual covert orienting is conserved across species despite different ecological niches. PMID- 11401013 TI - Interaction of olanzapine with fluvoxamine. PMID- 11401014 TI - Lung cancer due to passive smoking--a review. AB - OBJECTIVES: Even from the scientific literature it is difficult to conclude whether the increased risk of lung cancer due to exposure to environmental tobacco smoke (ETS), as reported in many epidemiological studies, is based on sound data from reliable studies, or rather on passionate assertions derived from unsound investigations. To shed some light on this matter the differences between cigarette mainstream smoke (MS)--inhaled by the smoker- and ETS--inhaled by everyone exposed-, the concentration of ETS under real life conditions, the internal dose of toxic compounds due to ETS exposure, and the risk of lung cancer as found in epidemiological studies are discussed. RESULTS: MS and ETS differ considerably in their physical, chemical and toxicological characteristics because of the different conditions under which they are generated, the dilution in air, and the degree of ageing. Based on toxicological data, a very low internal dose of potentially genotoxic compounds can be measured in people after ETS exposure. The epidemiological data suggest a slightly increased risk of lung cancer in non-smokers chronically exposed to ETS. However, it is equally well known, that none of these studies is free from bias and confounding effects. CONCLUSION: The average intake of toxic and genotoxic compounds due to ETS exposure is that low that it is difficult, if not impossible, to explain the increased risk of lung cancer as found in epidemiological studies. The uncertainty is further increased because the validity of epidemiological studies on passive smoking is limited severely by numerous bias and confounding factors which cannot be controlled for reliability. The question of whether or not ETS exposure is high enough to induce and/or promote the carcinogenic effects observed in epidemiological studies thus remains open, and the assumption of an increased risk of lung cancer due to ETS exposure is, at present, more a matter of opinion than of firm scientific evidence. PMID- 11401015 TI - Flow cytometric analysis of lymphocyte subpopulations in bronchoalveolar lavage fluid after repeated ozone exposure. AB - As known from studies in animal and human subjects, ozone can exert effects on the immune response including allergic sensitisation and allergen responsiveness. The objective of the present study was to assess the changes in lymphocyte subsets in bronchoalveolar lavage fluid (BALF) after single and repeated ozone exposures. Twenty-three healthy subjects underwent single exposures to 200 ppb ozone or filtered air (FA), as well as repeated exposures to 200 ppb ozone on four consecutive days, each during 4 h of intermittent exercise. Bronchoalveolar lavage was performed 20 h after the single exposure or the last of the repeated exposures. Lymphocytes were identified by sideward scatter and CD45 expression, and their subsets by eight different panels of antibodies. Checksums were calculated to assess the validity of the results. The percentage and the absolute number of lymphocytes, mostly comprising T-lymphocytes (CD2+; overall mean 98.8%), increased after single (P < 0.05; each), but not after repeated ozone exposure, compared with FA (7.4 vs 5.8 vs 6.5%; 680 vs 419 vs 301 x 10(3)). In addition, we observed small but statistically significant changes in the proportions of lymphocyte subpopulations. The percentage of CD4+ lymphocytes increased after single (P < 0.05) and repeated ozone exposure (P < 0.01), whereas the percentage of CD8+ cells decreased after repeated exposure (P < 0.05). The proportion of activated lymphocytes (CD25+) was elevated after repeated, compared with single, ozone exposure (P < 0.01), and the percentages of natural killer (NK) cells were decreased after both single (P < 0.05) and repeated (P < 0.01) exposures. Our data suggest that single but not repeated ozone exposures cause a change in absolute numbers of lymphocytes in BALF, whereas the proportions of lymphocyte subsets are affected by single as well as repeated exposures. PMID- 11401016 TI - Albumin, transferrin and alpha2-macroglobulin in bronchoalveolar lavage fluid following exposure to organic dust in healthy subjects. AB - OBJECTIVES: The purpose of the present study was to investigate leakage of plasma proteins in connection with the inflammatory airway reaction following exposure to dust in a pig house. Inhalation of swine-house dust causes intense inflammation with influx of inflammatory cells, predominantly neutrophils, into the airways. The aim of the study was to compare the concentration of three different proteins in bronchoalveolar lavage (BAL) fluid as markers for the inflammation. METHODS: In twenty healthy, non-allergic, non-smokers, not previously exposed to farm dust, BAL was performed approximately 2 weeks before and 24 h after 3 h of exposure to swine dust in a swine-confinement building. Differential cell count and protein concentration were assessed in BAL fluid. Albumin (66.5 kDa) and alpha2-macroglobulin (720 kDa) were quantified by the use of enzyme-linked immunosorbent assay (ELISA) techniques, and transferrin (80 kDa) by zone immunoelectrophoresis assay. The coefficient of variation for repeated protein measurements was <9%. RESULTS: alpha2-Macroglobulin concentration increased six-fold, from 68.0 (36.1-99.9) microg/l, mean (95% CI) before exposure to 411.2 (254.0-568.4) microg/l after exposure (P < 0.001). Transferrin and albumin increased from 19.7 (16.2-23.1) mg/l and 1.8 (1.4-2.2) mg/l, 2.6 and 1.9 times, respectively (P < 0.001). There was significant correlation between the exposure-induced increased protein levels in BAL fluid, although alpha2 macroglobulin was a better discriminator of pre- and post-exposure concentrations than were albumin and transferrin. There was a significant correlation between the exposure-induced BAL-fluid neutrophilia and the increase in alpha2 macroglobulin and transferrin, but not for albumin. This correlation was found only when pre- and post- differences, but not ratios, of plasma proteins were compared. CONCLUSIONS: The levels of plasma proteins increased in BAL fluid following exposure to swine-house dust. alpha2-Macroglobulin was a better marker of this plasma leakage than were albumin and transferrin. PMID- 11401017 TI - Mortality and cancer incidence among a population previously exposed to environmental cadmium. AB - OBJECTIVES: This paper evaluates the associations of previous exposure to environmental cadmium (Cd) and renal function with total mortality and cancer incidence. METHODS: The study population comprised 275 residents (aged 40-92 years at baseline) in a Cd-polluted area located on Tsushima Island, Nagasaki, Japan. In the study area, the dietary intake of Cd decreased because the soil of the Cd-polluted rice fields was replaced with new soil between 1980 and 1983. The mortality rate from 1982 to 1997 and cancer incidence from 1985 to 1996 were investigated. Standardized mortality and incidence ratios (SMR and SIR) were calculated by using regional reference rates. The associations of renal function and urinary Cd levels with total mortality and cancer incidence were evaluated with Cox regression models. RESULTS: The SMR for all subjects, and those with a urinary beta2-microglobulin (U-beta2M) concentration > or = 1,000 microg/g creatinine (Cr) and < 1,000 microg/g Cr was estimated at 90 [95% confidence interval (CI) 73-109], 138 (95% CI 101-183) and 66 (95% CI 49-87), respectively. After adjustment for age and other potential confounders, in men, serum beta2M (S beta2M) (> or = 2.3 mg/l) and in women, serum Cr (> or = 21.2 mg/ 100 ml), relative clearance of beta2M (> or = 21%) and U-beta2M (> or = 1,000 microg/g Cr), were associated with a significantly increased risk of mortality, with hazard ratios exceeding 2.0. After further adjustment for log(U-beta2M), the rate ratio of deaths associated with, in men, increased S-beta2M was 2.53 (95% CI 0.97 6.65) and, in women, increased serum Cr (S-Cr) concentrations was 2.75 (95% CI 1.24-6.14). Urinary Cd concentrations (> or = 10 microg/g Cr) were not significantly associated with mortality. The overall SIR of all malignant neoplasms was 71 (95% CI 44-107). CONCLUSIONS: These findings suggest that renal tubule dysfunction and a reduced glomerular filtration rate are predictors of mortality among persons previously exposed to environmental Cd. However, the results also suggest that overall mortality rates in Cd-polluted areas are not necessarily increased, because of the low mortality among those with no, or only slight, signs of low-molecular weight proteinuria. Overall cancer incidence may not be increased among residents in Cd-polluted areas. PMID- 11401018 TI - Levels of metals and organic substances in blood and urine of workers at a new hazardous waste incinerator. AB - OBJECTIVE: To assess baseline concentrations of a number of metals and organic compounds in blood and urine of 28 workers employed at a new hazardous waste incinerator (HWI), before operation of the plant. METHODS: Plasma analyses of hexachlorobenzene (HCB), polychlorinated biphenyls (PCB 28, 52, 101, 138, 153 and 180), benzene, toluene, ethylbenzene, m-xylene, and polychlorinated dibenzo-p dioxins and dibenzofurans (PCDD/Fs) were carried out. The urinary levels of 2,4- and 2,5-dichlorophenol (DCPs), 2,4,5- and 2,4,6-trichlorophenol (TCPs) and pentachlorophenol (PCP), as well as those of 1-hydroxypyrene (1-HP) were also measured. Beryllium (Be), manganese (Mn), mercury (Hg) and lead (Pb) concentrations were determined in total blood, while the levels of arsenic (As), cadmium (Cd), chromium (Cr), nickel (Ni) and vanadium (V) were measured in urine. RESULTS: The levels of benzene, toluene, ethylbenzene and m-xylene were under their respective detection limits. The concentrations of HCB, PCBs and PCDD/Fs in plasma of the HWI workers, chlorophenols (CLPs) and 1-HP urinary concentrations, as well as those of metals in blood and urine are shown. PCDD/F concentrations in plasma of the new HWI workers ranged between 13.4 and 84.0 pg international toxic equivalents (I-TEQ)/g lipid, with a geometric mean value of 24.4 pg I-TEQ/g lipid. DISCUSSION: PCDD/F levels, as well as the concentrations of PCBs were of the same order of magnitude as those recently obtained for non-occupationally exposed populations of Catalonia (Spain). No significant gender differences were found for the levels of HCB, PCBs and PCDD/Fs. Although the present results showed a notable variability in the levels of CLPs, geometric mean values of these compounds were similar or even lower than those considered as potential reference values. Metal concentrations in blood and urine were also of the same order of magnitude than previously reported metal levels in the same area. CONCLUSION: The present results should be useful in future surveys in which internal exposure of the HWI workers will be determined. PMID- 11401019 TI - Sick building syndrome in relation to building dampness in multi-family residential buildings in Stockholm. AB - OBJECTIVES: The aim was to study relationships between symptoms compatible with sick building syndrome (SBS) on one hand, and different indicators of building dampness in Swedish multi-family buildings on the other. METHODS: In Stockholm, 609 multi-family buildings with 14,235 dwellings were identified, and selected by stratified random sampling. The response rate was 77%. Information on weekly symptoms, age, gender, population density in the apartment, water leakage during the past 5 years, mouldy odour, condensation on windows, and high air humidity in the bathroom was assessed by a postal questionnaire. In addition, independent information on building characteristics was gathered from the building owners, and the central building register in Stockholm. Multiple logistic regression analysis was applied, and adjusted odds ratios (OR) were calculated, adjusted for age and gender, population density, and selected building characteristics. RESULTS: Condensation on windows, high air humidity in the bathroom, mouldy odour, and water leakage was reported from 9.0%, 12.4%, 7.7% and 12.7% of the dwellings, respectively. In total 28.5% reported at least one sign of dampness. All indicators of dampness were related to an increase of all types of symptoms, significant even when adjusted for age, gender, population density, type of ventilation system, and ownership of the building. A combination of mouldy odour and signs of high air humidity was related to an increased occurrence of all types of symptoms (OR = 3.7-6.0). Similar findings were observed for a combination of mouldy odour and structural building dampness (water leakage) (OR = 2.9 5.2). In addition, a dose-response relationship between symptoms and number of signs of dampness was observed. In dwellings with all four dampness indicators, OR was 6.5, 7.1, 19.9, 5.8, 6.1, 9.4, 15.0 for ocular, nasal, throat, dermal symptoms, cough, headache and tiredness, respectively. CONCLUSION: Signs of high air humidity, as well as of structural building dampness, are common in multi-family buildings in Stockholm. Reports of building dampness in dwellings is related to a pronounced increase of symptoms compatible with the SBS, even when adjusted for possible confounding by age, gender, population density, and building-related risk factors. PMID- 11401020 TI - Nasal symptoms and pathophysiology in farmers. AB - OBJECTIVES: Increased morbidity and mortality in lower airway diseases have been reported among farmers. The aim of this study was to assess upper airway problems in farmers. SUBJECTS AND METHODS: Twenty-five dairy farmers, 20 pig farmers, 21 grain farmers and 19 control subjects were studied, by use of questionnaire, skin prick test, dynamic spirometry, nasal inspection, acoustic rhinometry (before and after a decongestant) as well as by determinations of the olfactory threshold and nasal lavage (NAL) concentrations of eosinophil cationic protein (ECP), myeloperoxidase (MPO) and albumin. RESULTS: Compared with the control group, farmers had more complaints of work-related symptoms from the lower airways, and symptoms of smell impairment, and more often had nasal polyps and hyperaemia of the nasal mucosa. They also had higher levels of MPO in NAL (especially dairy farmers and pig farmers), and a tendency to more swollen nasal mucosa and lower olfactory threshold (especially grain farmers). CONCLUSIONS: The farmers had more pathological findings in their nasal mucosa, possibly indicating effects of allergens and irritants in their work environment. More studies are needed to evaluate work environment factors causing these pathological findings in farmers. PMID- 11401021 TI - Cognitive functions in workers exposed to toluene: evaluation at least 48 hours after removal from exposure. AB - OBJECTIVES: Long-term exposure to toluene may result in subtle impairment of cognitive functions. However, it is not clear whether this impairment is due to the presence of the solvent in the body or if it persists after its elimination from blood. The aim of this study is to compare cognitive functions between toluene-exposed workers (at least 48 h after removal from exposure) and non exposed workers. METHODS: Seventy-two workers exposed for at least 5 years to toluene (9 to 467 ppm) completed a questionnaire and psychometric tests. The results were compared with those of 61 non-exposed workers. An alveolar air sample was taken just before the tests to ensure the absence of toluene. RESULTS: Results of the vocabulary test were slightly better in exposed (correct words: 21 +/- 0.6) than in non-exposed workers (19 +/- 0.8) (P < 0.05). No differences were found for simple reaction time, digit symbol, digit span, continuous tracking test, color word and switching attention test. CONCLUSIONS: The results of this study do not support the notion of the persistence of cognitive effects of toluene after elimination of the solvent from blood. PMID- 11401022 TI - N-methylsuccinimide in plasma and urine as a biomarker of exposure to N-methyl-2 pyrrolidone. AB - OBJECTIVE: N-Methyl-2-pyrrolidone (NMP) is a selective and powerful organic solvent. The aim of this study was to investigate whether the NMP metabolite N methylsuccinimide (MSI) in plasma and urine can be used as a biomarker of exposure to NMP. METHODS: Six healthy subjects were exposed to 10, 25, and 50 mg NMP/m3 in an exposure chamber for 8 h. The air levels were monitored by XAD-7 solid sorbent sampling, and analysed by gas chromatography (GC). Plasma and urine were sampled for two days following the exposure, and the levels of MSI were analysed by GC with mass spectrometric detection. RESULTS: The concentration of MSI in plasma and urine rose during the exposure, and reached a peak at about 4 h after the end of the exposure. The concentration then decayed according to a one compartment model with a half-time of approximately 8 h. About 1% of the inhaled NMP was excreted in urine as MSI. There were very close correlations between the NMP air levels and, on the one hand, the MSI concentrations in plasma collected at the end of exposure (r = 0.98), or the urinary MSI concentration collected during the last 2 h of exposure (r = 0.96), on the other. CONCLUSIONS: MSI in plasma or urine is applicable as a biomarker of exposure to NMP. The concentration in plasma and urine mainly reflects the exposure over one day. PMID- 11401023 TI - Experiences on persistent organic pollutants under the Law concerning the Examination and Regulation of Manufacture, etc. of Chemical Substances, Japan, with references to biodegradation and bioaccumulation. AB - OBJECTIVES: The purpose of this report is to outline current regulations to control chemical environmental pollution in Japan, with special references to internationally defined 12 persistent organic pollutants (POPs). MATERIALS: Law concerning the Examination and Regulation of Manufacture, etc. of Chemical Substances [(LERCS); enacted in 1973] and related administrative activities of monitoring of the environment in Japan. RESULTS: Among the existing chemicals identified by the 1972 Chemicals Inventory, LERCS designates aldrin, chlordanes, DDT, dieldrin, endrin, HCB, PCBs, poly(Cln; n = 3 or more)-chlorinated naphthalene (PCNs) and bis(tributyltin) oxide (TBTO) as Class 1 specified chemical substances which are under strict regulation, such as prohibition of production, import, or use in principle. In addition LERCS designates 23 Class 2 specified chemical substances (including 13 tributyltin and seven triphenyltin compounds) for which notification of scheduled and past production, compliance with technical guidelines and compliance with labeling standards is requested. When compared with the 12 POPs, the designation covers most of them except for mirex and toxaphene, which have never been used in Japan. The regulation has been effective in reducing substantially the levels of the designated chemical substances (and therefore the 12 POPs except for dioxins and furans) in the general environment in Japan. Efforts are currently focused under a newly enacted law to reduce the emission of the two non-intentionally produced pollutants of dioxins and furans, especially from city waste incinerators, so that emission in 2002 should be 10% of that in 1997. CONCLUSION: Regulations to control chemical emissions have been effective in reducing POPs levels in the environment in Japan, and further efforts have been made under a new law to reduce the emission of dioxins, furans and co-planar PCBs. PMID- 11401024 TI - Targeting properties of an anti-CD16/anti-CD30 bispecific antibody in an in vivo system. AB - Bispecific antibodies are currently being used in clinical trials in increasing numbers in the areas of breast cancer, prostate cancer, non-Hodgkin's lymphoma and Hodgkin's lymphoma. We have previously performed two clinical trials in patients with Hodgkin's disease with an anti-CD30/anti-CD16 bispecific antibody and demonstrated a 30% response rate in a cohort of patients otherwise resistant to standard therapeutic modalities. However, no surrogate marker could be defined in these trials indicative of optimal antibody dosing/scheduling or predictive for favorable response. In order to evaluate accurately the potential biodistribution properties of bispecific antibody in patients, we have performed a detailed analysis of the binding properties and animal model in vivo characteristics of these constructs. For this purpose, the parental antibodies (anti-CD30 and anti-CD16) and the bispecific antibody (anti-CD30/anti-CD16) were radiolabeled with either 125I or 111In. Antibody integrity and binding properties after labeling were confirmed by Scatchard plot and Lindmo analysis. 111In labeled antibodies revealed superior targeting properties in a standard SCID mouse tumor model. Both the bivalent parental anti-CD30 monoclonal antibody and the monovalent anti-CD30/anti-CD16 bispecific antibody showed excellent uptake in CD30+ tumors which did not differ significantly between the two (maximum uptake 16.5%+/-4.2% vs. 18.4%+/-3.8% injected dose/gram tissue). The equivalent targeting properties of the bispecific antibody compared with the parental anti CD30 antibody encourages the further clinical development of this bispecific antibody, and might help to explain the clinical responses seen with this antibody so far in patients suffering from Hodgkin's disease. PMID- 11401025 TI - Generation of human anti-MUC3 IgG antibodies after in vitro immunization of naive peripheral blood B-lymphocytes. AB - It has been demonstrated that IgG antibodies can be generated to self-antigen peptides as well as against viral antigens by an antigen-specific in vitro immunization system of resting human peripheral B-lymphocytes. Using a synthetic peptide from the consensus variable tandem-repeat region of the MUC3 mucin (TSSITTTGTTSHSTPSP) as the B cell epitope, we immunized blood donor B-lymphocytes in vitro and tested for MUC3-specific antibodies by ELISA. After the primary activation step all antibodies were IgM. At the end of the secondary immunization step we obtained 1.8% (21/1138) of the cultures with IgG-switched antibodies. In a competitive inhibition ELISA using the MUC1, MUC2, MUC3, MUC4 and PIP2 peptides, only one culture (F8.1) gave satisfactory specific inhibition. Using this antibody in fluorometric studies, it stained cells from two colon carcinoma cell lines predominantly in the cytoplasm, whereas those from a breast cancer cell line stained predominantly the cell surface. In a preliminary immunohistological evaluation with formalin-fixed sections, the antibody appeared to moderately stain colon sections, but not breast sections or lymph node. This method of in vitro immunization may be a useful tool in generating IgG antibodies specific to self-antigens and could find applications in tumour targeting and immunotherapy. PMID- 11401026 TI - Expression of L-selectin and efficient binding to high endothelial venules do not modulate the dissemination potential of murine B-cell lymphoma. AB - The homing receptor L-selectin is essential for the migration of naive lymphocytes into peripheral lymph nodes. In contrast to naive lymphocytes, activated and memory cells down-regulate L-selectin and enter peripheral lymph nodes by an L-selectin-independent mechanism. In view of the concept that lymphomas present the malignant counterparts of normal lymphocytes at a defined stage of differentiation, it has been suggested that in contrast to lymphomas with a memory/activated cell phenotype, L-selectin is essential for dissemination of lymphomas that represent naive cells. 38C-13 is a murine B-cell lymphoma with an immature naive cell phenotype. 38C-13 cells express high levels of L-selectin and bind to lymph node high endothelial venules in an L-selectin-dependent manner. In this study we demonstrate that treatment of 38C-13 tumor-bearing mice with anti-L-selectin antibodies did not inhibit tumor dissemination to peripheral lymph nodes. Moreover, L-selectin-negative 38C-13 variant cells disseminated as efficiently as wild-type cells. Thus, in spite of its expression, L-selectin is not required and does not affect the metastatic potential of the tumor. L selectin of the malignant cells and of normal lymphocytes appears to be functionally different. Thus, whereas antibody cross-linking of L-selectin resulted in down-modulation of the receptor in normal lymphocytes, cross-linking had no effect on L-selectin expression in 38C-13 cells, suggesting that, in spite of comparable levels of surface expression in normal and malignant cells, L selectin may be functionally impaired in some malignant cells. PMID- 11401027 TI - Tributyrin enhances the cytotoxic activity of interleukin-2/interleukin-12 stimulated human natural killer cells against LS 174T colon cancer cells in vitro. AB - Tributyrin has been shown to be cytostatic to tumor cells by inducing differentiation and apoptosis. On the other hand, immunological NK cells can kill tumor cells, particularly when stimulated with interleukin-2 (IL-2) and/or interleukin-12(IL-12). However, little is known about whether and how both antitumor mechanisms act together, although in vivo such an interaction must exist. Here we demonstrate in vitro, that pretreatment of human LS 174T colon cancer cells with nontoxic concentrations of tributyrin augments the sensitivity to spontaneous NK cell activity two-fold. However, when NK cells have been activated with an optimized combination of IL-2 and IL-12, the immunocytotoxicity increases up to five-fold (from 14% to 70%), versus a 3.8-fold increase against untreated cancer cells. These effects are accompanied by increased IFN-gamma secretion and decreased TGF-beta1 secretion. Tributyrin is found to be a potent inducer of ICAM-1, LFA-3 and Fas on target cells corresponding to an increase of the FasL expression by IL-2/IL-12 on the effector cells. Our data suggest a synergistic link between induction of tumor cell differentiation and immunological defense mechanisms that may provide a rational basis for the improvement of clinical protocols, especially for colon cancer. PMID- 11401028 TI - Relationship between tumour morphology, antigen and antibody distribution measured by fusion of digital phosphor and photographic images. AB - Antibody-directed cancer therapy has achieved encouraging responses despite poor localisation in tumour. This discrepancy may be attributed to heterogeneity of antibody delivery within tumours: preferential localisation in the better perfused and more radio- and chemosensitive areas provides a therapeutic advantage. Antibody distribution depends upon the interactions of many complex mechanisms. We have started to investigate this by studying the single and combined influence of two tumour-associated parameters, morphology and antigen, on antibody distribution. Tumours were taken from mice at 24 and 48 h after 125I labeled anti-CEA antibody injection. Images of antibody distribution, antigen distribution and tumour morphology were acquired by radioluminography, radioimmunoluminography and digitisation of morphology, respectively. Image registration allowed correlation of pixel values of antibody distribution with corresponding values of antigen distribution and morphology. At 24 h there was little correlation between antibody and antigen distribution, but strong positive correlation between antibody distribution and morphology, with preferential localisation in viable tumour areas. Correlation between antibody distribution and morphology fell significantly between 24 and 48 h, while that between antibody and antigen distribution remained low. However, the combination of morphology and antigen distribution showed the largest influence on antibody distribution. This novel technique demonstrates potential for combining multi factor information in order to provide a greater understanding of antibody distribution in tumours, facilitating the optimisation of clinical treatments. PMID- 11401029 TI - Long-term treatment with low doses of interleukin-2 and interferon-alpha: immunological effects in advanced renal cell cancer. AB - PURPOSE: We aimed to determine the immunological effects of low doses of recombinant interleukin-2 (rIL-2) and recombinant interferon-alpha (rIFN-alpha) in patients bearing advanced renal cell carcinoma. METHODS: Twenty-seven patients received therapeutic cycles consisting of subcutaneous rIL-2 for 5 days per week and intramuscular rIFN-alpha twice weekly, for 4 consecutive weeks. The cycle was repeated indefinitely at regular 4-month intervals, for all patients. rIL-2 (1 x 10(6) IU/m2) was administered every 12 h on days 1 and 2 and once a day on days 3 5 of each week; rIFN-alpha (1.8 x 10(6) IU/m2) was given on days 3 and 5. In the enrolled patients, total and differential white blood cell counts, phenotypic analysis of some lymphocyte subsets, and soluble IL-2 receptor (sIL-2R), were investigated before and after each of the first six cycles of therapy (about 24 months of follow-up). RESULTS: The cycles of immunotherapy induced a significant increase of total lymphocytes (37%, P < 0.001), eosinophils (222%, P < 0.001), CD25+ cells (27%, P=0.004), sIL-2R (174%, P < 0.001) and natural killer (NK) cells (CD3-CD56+) (61%, P < 0.001); the subset that expresses CD56 with high density (CD56+ bright) expanded more (233%, P < 0.001) than the subset expressing the same marker with low density (CD56+ dimmer) (15%, P = 0.043). Unlike the previous subsets, the treatment decreased significantly T-lymphocytes with NK cell marker (CD3+ CD56+) (28%, P = 0.011). No significant differences of effectiveness were found among the subsequent treatment cycles, except for CD25+ cells and sIL-2R (P = 0.036 and P = 0.005, respectively): the increase induced by immunotherapy was maximum after the first cycle and decreased progressively thereafter. CONCLUSIONS: Long-term repeated cycles of low-dose immunotherapy induced repeated and significant expansion of one of the most important lymphocyte subsets for the non-MHC-restricted immune response to the tumour mass: CD3-CD56+ cells. PMID- 11401030 TI - Active idiotypic vaccination in a patient with biclonal follicular lymphoma. AB - Specific immunological responses to the idiotypic epitopes present in the surface immunoglobulin (Ig) of the clonal tumour population can be induced for active immunotherapy in patients with B-cell non-Hodgkin lymphoma (NHL). The clonality of the tumour cells should have important implications for the success of the implemented therapy. Here we report on the case of a patient enrolled in a protocol of active idiotypic immunotherapy in which previous cytofluorometric analysis showed a major IgM+, kappap+ population in the tumoral cell suspensions. However, sequence analysis of both tumour sample and tumour-derived hybrids revealed the presence of two unrelated clones that used different VH and VK gene segments. It was possible to obtain hybridomas secreting these two different IgM, kappap idiotypic proteins. The patient was immunised with a mixture of these two idiotypic Igs conjugated to keyhole limpet haemocyanin. Anti-idiotypic antibodies directed against both tumour-associated proteins were detected. This is the first case of anti-idiotypic therapy in a patient with a biclonal NHL. Our work calls attention to the question of clonality in the context of idiotypic vaccination in NHL patients. PMID- 11401031 TI - Human scFv antibody fragments specific for the epithelial tumour marker MUC-1, selected by phage display on living cells. AB - New anti-cancer agents are being developed that specifically recognise tumour cells. Recognition is dependent upon the enhanced expression of antigenic determinants on the surface of tumour cells. The tumour exposure and the extracellular accessibility of the mucin MUC-1 make this marker a suitable target for tumour diagnosis and therapy. We isolated and characterised six human scFv antibody fragments that bound to the MUC-1 core protein, by selecting a large naive human phage display library directly on a MUC-1-expressing breast carcinoma cell line. Their binding characteristics have been studied by ELISA, FACS and indirect immunofluorescence. The human scFv antibody fragments were specific for the tandem repeat region of MUC-1 and their binding is inhibited by soluble antigen. Four human scFv antibody fragments (M2, M3, M8, M12) recognised the hydrophilic PDTRP region of the MUC-1 core protein, which is thought to be an immunodominant region. The human scFv antibody fragments were stable in human serum at 37 degrees C and retained their binding specificity. For imaging or targeting to tumours over-expressing MUC-1, it might be feasible to use these human scFv, or multivalent derivatives, as vehicles to deliver anti-cancer agents. PMID- 11401032 TI - Signal transduction and Dictyostelium development. PMID- 11401033 TI - Meeting report: Molecular Parasitology XI, Woods Hole, USA, September 17-21, 2000. PMID- 11401034 TI - From parasites to life in space! PMID- 11401035 TI - Involvement of a 40-kDa glycoprotein in food recognition, prey capture, and induction of phagocytosis in the protozoon Actinophrys sol. AB - A 40-kDa glycoprotein (gp40) was identified as a Con A-binding adhesive substance of the heliozoon Actinophrys sol for immobilizing and ingesting prey flagellates. Isolation and partial characterization of gp40 showed that: 1) gp40 is a major Con A-binding protein of Actinophrys with a molecular weight of 40 kDa, and is stored in secretory granules called extrusomes; 2) gp40 was purified by Con A affinity chromatography, and the N-terminal amino acid sequence was determined as H2N-KVLK-FEDDFDTFDLQ; 3) prey flagellates became adhered to gp40-immobilized agarose beads; 4) phagocytosis of Actinophrys was induced against gp40 immobilized agarose beads; and 5) solubilized gp40 induced exocytosis of extrusomes and cell fusion of heliozoons. These results indicate that gp40 is a multi-functional secretory protein of Actinophrys, which is required in correct targeting of the heliozoon to food organisms as well as in self-recognition. PMID- 11401036 TI - A phylogenetic analysis of the SSU rRNA from members of the Plasmodiophorida and Phagomyxida. AB - The Plasmodiophorida and Phagomyxida are orders of zoosporic eukaryotes of uncertain phylogenetic affinities. We have obtained ribosomal DNA sequence information from several species in these groups. Phylogenetic analyses of these sequences have confirmed that they form a monophyletic grouping that clusters most closely with a rhizopod assemblage consisting of sarcomonads and chlorarachneans. The Phagomyxida sequences appear distant enough from those of the Plasmodiophorida to justify a separate order. The relationships between the Plasmodiophorida remain unclear, with species from the same genus showing significant degrees of divergence. PMID- 11401037 TI - The dynamics of filamentous structures in the apical band, oral crescent, fission line and the postoral meridional filament in Tetrahymena thermophila revealed by monoclonal antibody 12G9. AB - The ciliate Tetrahymena thermophila possesses a multitude of cytoskeletal structures whose differentiation is related to the basal bodies - the main mediators of the cortical pattern. This investigation deals with immunolocalization using light and electron microscopy of filaments labeled by the monoclonal antibody 12G9, which in other ciliates identifies filaments involved in transmission of cellular polarities and marks cell meridians with the highest morphogenetic potential. In Tetrahymena interphase cells, mAb 12G9 localizes to the sites of basal bodies and to the striated ciliary rootlets, to the apical band of filaments and to the fine fibrillar oral crescent. We followed the sequence of development of these structures during divisional morphogenesis. The labeling of the maternal oral crescent disappears in pre-metaphase cells and reappears during anaphase, concomitantly with differentiation of the new structure in the posterior daughter cell. In the posterior daughter cell, the new apical band originates as small clusters of filaments located at the base of the anterior basal bodies of the apical basal body couplets during early anaphase. The differentiation of the band is completed in the final stages of cytokinesis and in the young post-dividing cell. The maternal band is reorganized earlier, simultaneously with the oral structure. The mAb 12G9 identifies two transient structures present only in dividing cells. One is a medial structure demarcating the two daughter cells during metaphase and anaphase, and defining the new anterior border of the posterior daughter cell. The other is a post-oral meridional filament marking the stomatogenic meridian in postmetaphase cells. Comparative analysis of immunolocalization of transient filaments labeled with mAb12G9 in Tetrahymena and other ciliates indicates that this antibody identifies a protein bound to filamentous structures, which might play a role in relying polarities of cortical domains and could be a part of a mechanism which governs the positioning of cortical organelles in ciliates. PMID- 11401038 TI - Two most remarkable Amoeba men: Joseph Leidy (1823-1891) of Philadelphia and Eugene Penard (1855-1954) of Geneva. PMID- 11401039 TI - Grazing-associated infochemicals induce colony formation in the green alga Scenedesmus. PMID- 11401040 TI - Practice and policy of measuring quality of life and health economics in cancer clinical trials: a survey among co-operative trial groups. AB - BACKGROUND: Co-operative groups have played an important role in the advance of health-related quality of life (HRQL) research. However, definitions of the concept, criteria for selection of existing instruments and methods for data collection and interpretation remain poorly defined in the literature. A survey was conducted amongst the major cancer co-operative groups in order to gain a better understanding of their current policy and processes to ensure optimal HRQL data collection within cancer clinical trials. The topic of health economics was similarly addressed. METHODS: A written questionnaire was addressed to 16 major European and North American cancer co-operative groups. Eleven groups responded (response rate: 69%). however, one group could not provide information for the survey, thus ten questionnaires were available for analysis. RESULTS: The results from this survey among co-operative groups show that HRQL (more than health economics) is recognized as an important, although usually secondary, outcome measure in oncology trials. On the whole, co-operative groups have a rather flexible policy towards the inclusion of HRQL (and HE) into their clinical trials, and practice is very much on a case-by-case basis, but use standard practice guidelines and internal procedures is to ensure well-defined study protocols and enhance good quality studies. PMID- 11401041 TI - Assessing sexual functioning in patients with chronic disorders by using a generic health-related quality of life questionnaire. AB - This study was an assessment of sexual functioning by making cross-sectional surveys of patients aged 45 84 years with chronic disorders; 397 had diabetes, 248 had glaucoma and 1914 had chronic, stable, angina pectoris; 1669 came from a standard population sample (SPS). Sexual functioning was assessed on one scale from the Swedish health-related quality of life survey (SWED-QUAL), adapted from the medical outcomes study (MOS). Both sexual functioning in general and specific sexual items were studied. Values were adjusted for non-respondents and standardised to the Swedish population. Response rates in the samples were 54-64% among men and 22-41% among women, decreasing with age for women. General sexual functioning decreased with age and was most apparent in patients with diabetes and angina pectoris, while glaucoma patients reported better results than the SPS. Loss of male erectile function was as common in diabetes (30%) as in angina pectoris (29%) and significantly higher than in the SPS (20%) (p < 0.001). Besides age, significant factors for erectile dysfunction were, in diabetes, the presence of microvascular complications, treatment with long-acting nitroglycerine and psychiatric disease, and in angina, the presence of diabetes and peripheral artery disease. PMID- 11401042 TI - Multiple chronic health problems are negatively associated with health related quality of life (HRQoL) irrespective of age. AB - PURPOSE: To examine HRQoL measured by EORTC QLQ-C30 with respect to an increasing number of self-reported chronic health problems in the general Swedish population and to study the association between HRQoL, chronic health problems and age, gender, income, marital status and employment status. METHOD: A postal survey among a large random sample of 4000 adults aged 18-79 years. The study material contained EORTC QLQ-C30 core questionnaire supplemented by a sociodemographic questionnaire including questions about 13 chronic health problems of which four categories, 'No', 'Few', 'Some' or 'A lot of chronic health problems were constructed. RESULTS: Multiple chronic health problems were significantly associated with reduced HRQoL. The increased number of chronic health problems was also associated with age. When the number of chronic health problems was accounted for, the influence of age diminished. Low income and unemployment were associated with greater decline in HRQoL with respect to increasing number of problems among the respondents in working age. CONCLUSION: The impact of increased number of chronic health problems had varying consequences in different age groups. Moreover, sociodemographic and economic factors showed to interact differently with chronic health problems and HRQoL in various age groups. It appears from our results that an assessment and a careful consideration of these factors will be valuable in order to facilitate the interpretation of the effects of cancer and treatment on long-term HRQoL of cancer patients. PMID- 11401043 TI - The relationship between asthma severity, family functioning and the health related quality of life of children with asthma. AB - This study compared the health-related quality of life (HRQL) of 236 children with mild or moderate/severe asthma with that of a large representative sample of children in the general community. The study also examined the relationship between the HRQL of children with asthma and their demographic characteristics, asthma severity and family functioning. Children with asthma had a significantly poorer HRQL than other children in the community. Amongst the children with asthma, parents reported that children living in single-parent families had poorer physical health, mental health and social functioning than children in two parent families. There was a significant relationship between the mental health of children with asthma and family functioning but no significant relationship between their physical health and family functioning. These findings suggest that the domains comprising the HRQL of children with asthma are related to both disease and non-disease factors. A better understanding of these relationships will facilitate the development of new interventions to help children with asthma. PMID- 11401044 TI - Functional abilities and continence: the use of proxy respondents in research involving older people. AB - As the use of proxy respondents is sometimes necessary in research involving older or disabled people it is important to assess the impact of this on research results. This study examined the concordance of proxy responses and index responses on questions concerning functional abilities and continence. Index respondents were women aged 65 years and older who were interviewed as part of a larger study of outcomes of fractured neck of femur, at 5 days post surgery. They were asked to nominate a proxy respondent who was interviewed using the same questions within a few days. Responses of proxy and index respondents were compared using percentage agreement and Kappa statistics. On questions regarding functional ability, proxy responses were found to be more reliable for personal care activities than for instrumental activities of daily living. This may be a result of questions concerning instrumental activities being somewhat more ambiguous and open to individual interpretation. Item non-response was low for all questions thus resulting in little missing data for proxy respondents. There was a tendency for more distant relationship and contact to produce better agreement, which is contrary to previous findings. Proxy responses were biased in the direction of an overestimation of functional incapacity and so researchers should be cautious in combining data from proxy and index respondents. Concordance was good for questions concerning urinary and faecal incontinence although non-response was higher than for functional ability questions. Concordance was not as great for more detailed questions concerning the timing and frequency of incontinence as these used graded response options, rather than simple yes/no responses. PMID- 11401045 TI - Assessing health-related quality of life in cataract patients: the relationship between utility and health-related quality of life measurement. AB - The relationship between health-related quality of life (HQL) measures and patient preference for their health status was studied. Study subjects consisted of 132 patients at four hospitals who were scheduled for cataract surgery. Generic and disease-specific health status measures were determined in study subjects. The Medical Outcomes Study Short-form 36 (SF-36) item health status instrument was used to measure generic health status and the Visual Function 14 (VF-14) item visual health status instrument was used as the disease-specific health measure. Preference for general health and visual health was measured by assessing utilities assigned by patients to certain health states. Utilities assigned for general health were correlated with all categories of the SF-36 and VF-14 scores. Utilities assigned for visual health were correlated with four categories of the SF-36 (role limitation due to emotional health, general health, physical functioning, and vitality) and VF-14 scores. Utilities assigned for visual health were more strongly correlated with VF-14 scores than generic measures of health. Verbal ratings for visual health were correlated with Snellen visual acuity (SVA) (r = 0.20), utilities assigned for visual health (r = 0.58), VF-14 scores (r = 0.74), all categories of the SF-36 (r values ranging from 0.21 to 0.28), utilities assigned for general health (r = 0.19), and verbal ratings for general health (r = 0.29). Utility measures and verbal ratings for general and visual health were shown to be appropriate HQL measures. These measures were strongly correlated with other established generic and disease-specific health measures and should be included in the array of health status measures. PMID- 11401046 TI - Re-validation and shortening of the Functional Assessment of Anorexia/Cachexia Therapy (FAACT) questionnaire. AB - PURPOSE: The original Functional Assessment of Anorexia/Cachexia Therapy (FAACT) was designed to measure general aspects of quality of life (QOL) as well as specific anorexia/cachexia-related concerns. Our primary purpose was to reduce the number of anorexia/cachexia subscale items in a manner that either retains or improves reliability, validity and precision. METHODS: The FAACT was administered using an interactive computer program that allowed immediate entry of the data. A total of 213 patients were recruited. RESULTS: A combined empirical and conceptual approach led to the reduction of the anorexia/cachexia subscale (A/CS) from 18 to 12 items. A 26-item trial outcome index (TOI) combining physical well being (PWB), functional well-being (FWB), and the A/CS-12 was highly reliable and sensitive to change in performance status rating (PSR). We found that PWB, FWB, and A/CS-12 subscales performed differently. Specifically, PWB and FWB scores decreased in patients whose (PSR) worsened. However, although A/CS-12 scores were responsive to change in PSR over time, average A/CS-12 scores of all patients, even those whose PSR worsened, improved over the course of treatment. CONCLUSIONS: Elimination of six items from the anorexia/cachexia subscale of the FAACT was accomplished without loss of internal consistency or sensitivity to change in performance status. The A/CS-12 subscale provides unique, important information not captured by a generic chronic illness questionnaire. PMID- 11401047 TI - The EORTC quality of life questionnaire-head and neck 35 in Italian laryngectomized patients. European organization for research and treatment of cancer. AB - The aim of this study was to test the validity and reliability of the European organization for research and treatment of cancer (EORTC) quality of life questionnaire (QLQ)-head and neck (H&N) 35 in Italian laryngeal cancer patients. The original questionnaire was developed by the EORTC quality of life (QoL) study group and tested in H&N cancer patients from Norway, Sweden, and the Netherlands. The Italian translation of the questionnaire used in this study was made by a team of the CRO, National Cancer Institute, using a double-back translation method between independent translators. The translated EORTC QLQ-H&N35 was given to 99 patients with H&N cancer who had undergone total laryngectomy 1-26 years before and had been then treated with radiotherapy and, in some cases, chemotherapy. The questionnaire was re-administrated to 33 patients after 1 month to test its stability over time. It was structurally made up of seven scales (pain, swallowing, sense, speech, social eating, social contact, and sexuality) and 11 single items that considered the most important clinical aspects characterizing the QoL in H&N cancer patients. The statistical analysis of the indexes of validity and reliability confirmed the results obtained with other linguistic versions of the questionnaire. Our Italian version of the EORTC QLQ H&N35 proved to be a statistically valid instrument to assess QoL in laryngectomized patients. PMID- 11401048 TI - Comparative studies of inhaled corticosteroids in asthma. PMID- 11401049 TI - The reproducibility and sample size requirements of exercise-induced bronchoconstriction measurements. AB - Dry air exercise challenges are frequently used to screen medications that have potential utility in the management of exercise-induced bronchoconstriction (EIB). The purpose of this study was to determine the reproducibility of three outcome measurements made using such challenges, and sample size requirements for drug evaluation studies based on these outcomes. Forty adult subjects with asthma, who tested positively on a screening exercise challenge, were subjected to two further identical challenges, separated by 1 to >35 days. Outcome measurements included the maximum per cent fall in forced expiratory volume in one second (FEV1), after exercise (% fallmax), and the area under the per cent fall in FEV1/time curve for 30 min (AUC30) and 60 min (AUC60) after exercise. The reproducibility of these outcomes, as assessed by intraclass correlation coefficients was 0.72, 0.53 and 0.35 for % fallmax, AUC30 and AUC60 measurements, respectively. The sample size requirements to demonstrate an attenuation of EIB equivalent to a 50% reduction in % fallmax was 9, 14 and 19 subjects for the % fallmax, AUC30 and AUC60 responses, respectively (90% power). It is concluded that the maximum percentage fall in forced expiratory volume in one second has greater reproducibility and results in greater power in clinical trials than area under the curve measurements. Sample size calculation curves are provided which may be used in study design and interpretation of published studies. PMID- 11401050 TI - Is overall asthma control being achieved? A hypothesis-generating study. AB - The efficacy of asthma therapy is traditionally measured using single end-points. In contrast, the aim of therapy is to achieve overall control, defined by management guidelines as achieving a number of treatment goals. These goals reflect expert opinion, rather than being evidence based. The objective of this study was to determine whether guideline-defined asthma control is achievable. Eight studies of salmeterol/fluticasone propionate combination therapy were analysed using three asthma control measures of varying stringency, derived from the guideline goals. For each measure, only patients meeting all goals were classified as controlled. The analysis demonstrated that asthma control, as defined by management guidelines, can be achieved. For a given therapy, similar proportions of patients achieved control irrespective of disease severity, suggesting that outcome expectations should not be reduced for patients with more severe disease. Substantially more patients achieved the target values for individual goals than achieved overall control, indicating that reliance on individual end-points is likely to result in significant overestimation of true control. The findings of this hypothesis-generating study should be prospectively tested. Future research will include a randomized controlled study designed to assess the proportion of patients able to achieve overall control of asthma when treatment is titrated appropriately. PMID- 11401051 TI - Prevalence of nasal symptoms and their relation to self-reported asthma and chronic bronchitis/emphysema. AB - Little information is available on associations between rhinitis and chronic bronchitis/emphysema (CBE). Self-reported upper airway symptoms, asthma, and CBE were examined in 12,079 adults living in southern Sweden. The response rate was 70% (n=8,469), of whom 33% reported significant nasal symptoms: a blocked nose was reported by 21%; sneezing by 18%; nasal discharge by 17%; and thick yellow nasal discharge by 5.7%. Nasal symptoms and combined nasal and self-reported bronchial disease were generally more common among smokers than nonsmokers. There was little overlap between asthma and CBE, but 46% of those with asthma and 40% of those with CBE had significant nasal symptoms. Best predicting factors (odds ratios >3) for asthma and CBE were nasal symptoms due to exposure to animals and damp/cold air, respectively. One-third of an adult, southern Swedish population, had significant allergic and/or nonallergic nasal symptoms. Nasal symptoms were frequently found to coexist with both asthma and chronic bronchitis/emphysema, suggesting that pan-airway engagement is common in both diseases. Differing associations between types of nasal symptoms and allergic and irritant triggers of nasal symptoms, with regard to asthma and chronic bronchitis/emphysema, emphasize the different natures of these bronchial diseases. PMID- 11401052 TI - The effect of sulphur dioxide exposure on indices of heart rate variability in normal and asthmatic adults. AB - Sulphur dioxide (SO2) is an important air pollutant and causes bronchoconstriction in normal and asthmatic adults. This paper has explored the autonomic consequences of SO2 exposure using the spectral analysis of heart rate variability. Electrocardiogram recordings were made in 12 normal and 12 asthmatic adults undergoing pollutant exposures. Exposures were of a 1 h duration, double blind, in random order, > or = 2 weeks apart and included air and 200 parts per billion SO2. Spectral analysis of R-R intervals was performed. SO2 exposure was associated with an increase in total power (TP) and high (HF) and low frequency (LF) power in the normal subjects, and a reduction in these indices in the subjects with asthma. The difference in TP with SO2 exposure compared to air was +1730 ms2 in the normal group and -1021 ms2 asthmatic group (p<0.003). For HF the respective values were +964 ms2 and -539 ms2 (p=0.02) and for LF, +43 7 ms2 and 57 2 ms2 (p=0.01). No change in lung function or symptoms was observed in either group. This suggests that SO2 exposure at concentrations which are frequently encountered during air pollution episodes can influence the autonomic nervous system. This may be important in understanding the mechanisms involved in SO2 induced bronchoconstriction, and of the cardiovascular effects of air pollution. PMID- 11401053 TI - Pulmonary gas exchange responses to histamine and methacholine challenges in mild asthma. AB - Histamine (HIST) produces greater changes in bronchial and pulmonary vasculature, and so may produce more gas exchange abnormalities, than methacholine (MTH) after inhalational challenge. The goals of this study were to compare the effects of HIST and MTH challenge on pulmonary gas exchange in patients with mild asthma at an equivalent degree of bronchoconstriction. Eleven patients were studied (mean+/ SEM age, 22+/-1 yr; forced expiratory volume in one second (FEV1), 91+/-5% pred) using a randomized, double-blind cross-over design. Respiratory system resistance (Rrs), arterial blood gases, and ventilation-perfusion distributions were measured before and after HIST/MTH challenges when cumulative doses caused a 30% fall in FEV1. Compared with baseline, HIST and MTH provoked similar moderate to severe increases in Rrs (p<0.005 each), and mild to moderate decreases in arterial oxygen tension (Pa,O2) due to ventilation-perfusion abnormalities (dispersion of pulmonary blood flow -log SDQ-, 0.40+/-0.03-0.71+/-0.08 and 0.47+/ 0.04-0.89+/-0.06; normal values <0.60-0.65), respectively, similar to those shown in mild to moderate acute asthma, without differences between them. For the same degree of airflow obstruction, both histamine and methacholine bronchoprovocations induce, in patients with mild asthma, very similar disturbances in ventilation-perfusion distribution and respiratory system resistance, suggesting similar mechanisms of airway narrowing. PMID- 11401054 TI - Effect of whole-body x-irradiation on antigen-induced airway response in sensitized guinea pigs. AB - The objectives of this study were to test the hypothesis that x-irradiation inhibits the late asthmatic response (LAR) without influencing the early asthmatic response (EAR) and to examine the mechanism of the inhibitory effect. Twenty sensitized guinea pigs were irradiated at a dose of 8 Gy. The next day, one-half of the animals were injected intravenously with spleen cells (2 x 10(8)) collected from unirradiated sensitized guinea pigs, whilst the other half were injected with vehicle only. Ten additional unirradiated sensitized guinea pigs also received vehicle only. Antigen inhalation challenge took place two days later. Pulmonary resistance was measured for 6 h after antigen exposure, and bronchoalveolar lavage and lung fixation were then undertaken. The area under the percentage pulmonary resistance curve 2-6 h after allergen inhalation was used for analysis of the LAR, while the maximal percentage change in pulmonary resistance was used for analysis of the EAR. Irradiation abolished the LAR (364.4+/-49.4 versus 62.8+/-10.4) without inhibiting the EAR (229.3+/-27.2 versus 278.7+/-40.2) and significantly inhibited the accumulation of eosinophils and lymphocytes in the airways. Transfer of spleen cells restored the LAR (334.4+/ 66.8) and the recruitment of cells to the levels seen in unirradiated sensitized guinea pigs. In addition, transfer of only CD4+ T-lymphocytes separated from the spleen cells restored the LAR (439.4+/-62.1) and the cell infiltration into the airways. These inhibitory effects of x-irradiation were due to decreases in numbers of CD4+ T-lymphocytes. PMID- 11401055 TI - Serum procalcitonin in pneumococcal pneumonia in children. AB - Serum procalcitonin (PCT), a marker of bacterial infection, was measured in children with pneumonia to examine whether PCT can be used to screen pneumococcal (PNC) from viral pneumonia. The number of patients was 132; mean age 3.0 yrs, and 64% were males. In all cases, pneumonia was radiologically confirmed, being alveolar in 46 and interstitial in 86 cases. The aetiology of infection was studied by a panel of serological tests for PNC, for five other respiratory bacteria and for seven common respiratory viruses. PNC infection was found in 25, mixed viral-PNC infections in 13 and viral infection in 17 cases. In general, serum PCT was not associated with the type or aetiology of pneumonia. PCT values were >1.0 mg.L(-1) in 40% of PNC cases, as compared to 12-15% in viral or mixed cases, respectively (p<0.05). PCT values were significantly higher in >2 yrs old children than in younger ones. The cut-off limits of 0.5 ng.mL(-1), 1.0 ng.mL(-1) and 2.0 ng.mL(-1) were tested for screening between PNC and viral pneumonia. The highest sensitivity of 55% was found at the 0.5 ng.mL(-1) cut-off level, whereas the highest specificity of 88% was reached at the level of 1.0 ng.mL(-1). The likelihood ratios, however, were far from optimal for both the positive and negative results. Although marginally higher in pneumococcal pneumonia than in viral pneumonia, serum procalcitonin cannot be used to discriminate between these two types of pneumonia. PMID- 11401056 TI - Pulmonary sarcoidosis in children: a follow-up study. AB - Progression of pulmonary sarcoidosis in children remains poorly documented. The aim of this work was to gather follow-up information on pulmonary outcomes in children with sarcoidosis and to obtain data of relevance to a discussion of the optimal length and regimen of glucocorticoid therapy. In the present study, the authors experience of pulmonary sarcoidosis in 21 children referred to the paediatric pulmonary department over a 10-yr period is reported with a documented follow-up of at least 4 yr. Evaluation of the disease during the follow-up included analysis of clinical manifestations, chest radiographs, pulmonary function tests with measurements of the vital capacity (VC), dynamic lung compliance (CL,dyn), lung transfer for CO (TL,CO), and arterial blood gases, as well as bronchoalveolar lavage (BAL) with determination of total and differential cell counts. After initial evaluation the decision was a careful observation of four children without therapy. Corticosteroid treatment was initiated in 17 children. Analysis of results indicated that after 6-12 months of treatment most clinical manifestations of the disease and chest radiograph abnormalities disappeared, and beneficial effects on VC and TL,CO were apparent. After 18 months of steroids no benefit on pulmonary function tests could be noticed, with mainly persistence of alterations of CL,dyn. Results of BAL studies documented the presence of an alveolitis with increased lymphocyte populations throughout the follow-up. Relapses were observed in four children during tapering of prednisone; they were not reported after discontinuation of steroid therapy. Taken together data obtained in the presented population can lead to the following suggestions for the management of pulmonary sarcoidosis in children. BAL should be performed at the initial evaluation to document alveolitis; however, nothing seems to be gained from repeating this investigation during follow-up in the absence of specific reasons. Once the decision to initiate glucocorticoid therapy is made, 18 months may be a reasonable treatment duration. Discontinuation of therapy can be decided even if the pulmonary function tests remain abnormal, but the child should then be carefully monitored for a relapse. PMID- 11401057 TI - Nosocomial bacterial pneumonia in human immunodeficiency virus infected subjects: incidence, risk factors and outcome. AB - The presented study examined the incidence, risk factors and outcome of nosocomial bacterial pneumonia (NBP) in human immunodeficiency virus (HIV) infected subjects. Forty-two cases of NBP were ascertained by a 5-yr prospective surveillance and were matched to 84 controls. NBP incidence was 10.8 per 10,000 hospital patient-days. In particular, the incidence of NBP was 13.9 per 10,000 patient-days in the period 1994-1996 and 5.6 per 10,000 patient-days in the period 1997-1998 (p=0.01). By using regression analysis, predictors for developing NBP were an increasing value of Acute Physiology and Chronic Health Evaluation (APACHE) III score (p<0.01) and the presence of acquired immune deficiency syndrome (AIDS)-related central nervous system (CNS) diseases (p=0.01). The additional hospital stay attributable to NBP was 15 days. The attributable mortality rate was estimated to be 29%. Nosocomial bacterial pneumonia is more common in patients with advanced human immunodeficiency virus infection, high Acute Physiology and Chronic Health Evaluation III score and central nervous system diseases. Although the incidence of nosocomial bacterial pneumonia, as well of other opportunistic infections, decreased considerably in the era of highly active antiretroviral therapy, it still represents an important cause of mortality. PMID- 11401058 TI - Sparfloxacin in the treatment of drug resistant tuberculosis or intolerance of first line therapy. AB - Patients with multiresistant tuberculosis (TB) and patients with intolerance of first line antituberculosis drugs present a major treatment problem. Sparfloxacin is highly active against mycobacteria, but the use is restricted by side effects and the contribution to antituberculosis therapy is unclear. A prospective study has therefore been performed to analyse the efficacy and tolerability of sparfloxacin in cases of resistant TB or intolerance of first line therapy. Between April 1993 and April 1999, 30 TB patients (28 with pulmonary TB and two with lymph node TB) were treated with combinations of sparfloxacin and at least two other drugs at the Chest Hospital Heckeshorn, Berlin. Sixteen patients were infected by resistant mycobacteria (one single drug resistance (SDR), one polyresistance, and 14 multidrug resistances (MDR); 14 males (age range 23-53 yrs), 2 females (68-74 yrs)). Twelve patients (11 males, one female, 27-80 yrs) had not tolerated first line antituberculosis drugs. Two additional male patients had continuous proof of Mycobacterium tuberculosis in sputum without resistance during therapy The duration of sparfloxacin therapy during hospitalization ranged 2.5-4 months. Twenty-five patients completed therapy and were cured according to this study's definition. Although sparfloxacin was generally well tolerated, five mild phototoxic reactions and six moderate prolongations of the electrocardiographic QT-interval (30-40 ms compared to baseline < or = 450 ms) were registered without clinical symptoms in the patient group. In summary, sparfloxacin proved an effective and safe alternative antituberculosis drug for complicated tuberculosis. PMID- 11401059 TI - Oxygen desaturation on the six-minute walk test and mortality in untreated primary pulmonary hypertension. AB - There are no reliable predictors of mortality in primary pulmonary hypertension (PPH). This study assessed whether exercise oxygen desaturation and distance achieved during a six-minute walk are associated with mortality in moderately symptomatic patients with PPH. Thirty-four patients with PPH underwent a pretreatment six-minute walk test, and an invasive haemodynamic assessment of pulmonary vasodilator reserve, to select the best treatment option (epoprostenol in 27 and nifedipine in 7). Median follow-up was 26 months (12 months for the nonsurvivors was 26%), and median survival, >46 months by Kaplan-Maier estimate. The mean+/-SD distance walked was 275+/-155 m and reduction in arterial oxygen saturation (Sa,O2) at maximal distance (deltaSa,O2) was 8.4+/-4.5%). A distance < or =300 m increased mortality risk by 2.4, and a deltaSa,O2 of > or = 10% increased mortality risk by 2.9. Only Sa,O2 at peak distance, deltaSa,O2 and pulmonary vascular resistance (PVR) were related to mortality. After adjusting for PVR, there remained a 27% increase in risk of death for each per cent decrease in Sa,O2. The six-minute walk distance and exercise oxygen saturation may be helpful in selecting patients with primary pulmonary hypertension for whom transplant listing is appropriate. PMID- 11401060 TI - The influence of fractional pulse pressure on the outcome of pulmonary thromboendarterectomy. AB - Although pulmonary thromboendarterectomy is an effective modality for the treatment of chronic thromboembolic pulmonary hypertension (CTEPH), the mortality in patients with severe haemodynamic disease is still high. Recently it was reported that fractional pulse pressure (pulmonary arterial pulse pressure/mean pulmonary arterial pressure) was higher in CTEPH than in primary pulmonary hypertension (PPH). It was hypothesized that fractional pulse pressure might be low in CTEPH with inaccessible distal thrombi and/or secondary pulmonary hypertensive change, resulting to the high operative mortality. To determine the influence of fractional pulse pressure to the outcome of surgery, 32 patients with CTEPH who had thromboendarterectomy between 1985 and 1998 were studied. Pulmonary haemodynamics and fractional pulse pressure were compared between survivors (n=26) and nonsurvivors (n=6) postoperatively. Those parameters in PPH (n=18) and large vessel pulmonary arteritis (n=6) were also analysed. Fractional pulse pressure in CTEPH (1.23+/-0.21) was significantly higher than in PPH (0.93+/-0.22; p=0.0017) and lower than in pulmonary arteritis (1.69+/-0.32; p=0.03). Fractional pulse pressure in survivors (1.26+/-0.21) was significantly higher than in nonsurvivors (1.06+/-0.16; p=0.03). Fractional pulse pressure is a significant predictor for mortality in patients with high pulmonary vascular resistance >1100 dynes.sec.cm(-5). To conclude fractional pulse pressure in addition to pulmonary vascular resistance might be useful in predicting for the outcome of surgery, especially in patients with severe haemodynamic impairment. PMID- 11401061 TI - Prognostic significance of p53 and bcl-2 abnormalities in operable nonsmall cell lung cancer. AB - The association of p53 abnormalities and bcl-2 protein expression with clinical data and prognosis in 102 patients with resected nonsmall cell lung cancer (NSCLC) was investigated. Deoxyribonucleic acid analysis of exons 5-8 of the p53 gene showed mutations (p53-M) in 47% of resected NSCLC, serum p53 antibodies (p53 Abs) were detected in 25%, p53 protein overexpression (p53-PE) in 54%, and bcl-2 protein overexpression (bcl-2-PE) in 48%. A statistically significant association was found between p53-PE, serum p53-Abs and the presence of a p53 gene alteration. No significant associations were found between results of the p53-M, p53-Abs, bcl-2-PE tests and clinicopathological parameters. In the case of the p53-PE test there were significantly fewer positive results for adenocarcinoma than for squamous cell carcinoma and large cell carcinoma. Survival analysis showed that both p53 abnormalities and negative staining for bcl-2, when analysed separately, were associated with poor overall survival. In a multivariate analysis, only the positive result of the p53-M test remained an independent, statistically significant, unfavourable prognostic factor for survival. When the p53 mutation test was removed from the model, positive results of the p53-PE test and the p53-Abs test became statistically significant, unfavourable prognostic factors. To conclude, among p53 and bcl-2 abnormalities, only p53 gene mutations seem to have a strong and independent effect on prognosis. When deoxyribonucleic acid sequence information is not available, p53 protein expression and the presence of p53 antibodies in serum may be used to obtain important prognostic information. PMID- 11401062 TI - Prognostic significance of blood coagulation tests in lung cancer. AB - Previous studies have shown that activation of coagulation has an impact on the clinical course of lung cancer. This study was carried out to assess the potential prognostic significance of platelet count (P), prothrombin time (PT), partial thromboplastin time (PTT), anti-thrombin III (AT-III), fibrinogen (F), D dimer (DD), factor II (F-II), factor VII (F-VII), factor X (F-X), protein C clotting (PCC), plasminogen (PL), and antiplasmin (AP) in 343 consecutive new lung cancer patients. A set of 32 anthropometric, clinical, physical, laboratory, radiological, and pathological variables was recorded prospectively for all patients. Patients were carefully followed-up, and their subsequent clinical course recorded. The most frequent abnormalities were of DD, F, and AT-III followed by F-VII, F-X, and F-II. Among the 12 clotting variables, the strongest relationships were those of F-II and F-X (Spearman rank (rs)=0.565), PT and F-VII (rs=0.562), F-VII and F-X (rs=0.514), PL and AP (rs=0.515), F and P (rs=0.490), AT-III and PCC (rs=0.476). Univariate analyses of survival showed that prolonged PT (p<0.043), and abnormally elevated DD (p<0.003), F (p<0.031), and P (p<0.047) were all associated with a poor prognosis. The multivariate model, however, did not confirm the prognostic significance of the coagulation factors. The results show subclinical activation of blood coagulation in lung cancer patients with early disease. In addition, clotting activation is confirmed as a predictor of survival, although not independently of other prognostic factors. PMID- 11401063 TI - Diaphragm fatigue during exercise at high altitude: the role of hypoxia and workload. AB - The effect of high altitude (HA) on exercise-induced diaphragm fatigue in normal subjects was examined. Eight normal subjects completed an incremental exercise test at sea level (SL) and at 3,325 m. Before (baseline), during, and after exercise (recovery), maximal transdiaphragm pressure (Pdi,sniff), breathing pattern, and diaphragmatic effort (PTPdi) were measured. Arterialized blood lactate was measured at baseline and during recovery. At maximal exercise (WRmax) Pdi,sniff fell to 72% and 61% of baseline at SL and HA respectively, recovering to baseline in 60 min at SL, and >60 min at HA. At the 5th min of recovery, circulating lactate was six-fold and seven-fold baseline at SL and HA, respectively. The time course of circulating lactate recovery was as for Pdi,sniff. At WRmax PTPdi was 80.74+/-9.87 kPa.s(-1) at SL and 64.13+/-8.21 kPa.s(-1) at HA. HA WRmax compared to isowork rate, SL data showed a lower Pdi,sniff (8.90+/-0.68 versus 11.24+/-0.59 kPa) and higher minute ventilation (117+/-11 versus 91+/-13 L.min(-1)), PTPdi being equal. To conclude, in normal subjects hypoxia-related effects, and not an increase in diaphragm work, hastens exercise-induced diaphragm fatigue and delays its recovery at high altitude compared to sea level. PMID- 11401064 TI - Diaphragmatic angiogenic growth factor mRNA responses to increased ventilation caused by hypoxia and hypercapnia. AB - This study investigates the effect of increased ventilation on the expression of messenger ribonucleic acid (mRNA) levels of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and transforming growth factor beta1 (TGF-beta1) in the diaphragm of intact, awake, spontaneously breathing rats, compared with responses in paralysed, mechanically-ventilated animals at similar blood gas and ventilatory levels. Four groups of intact, rats were studied in a body box, each group breathing one of four gases: room air, 12% oxygen (O2), 5% carbon dioxide (CO2), or 12% O2+5% CO2 for 1 h. Another 4 groups of paralysed, mechanically-ventilated animals were matched for arterial blood gas and ventilatory level. The results showed that VEGF mRNA abundance was increased three-fold and that of bFGF 1.5-fold when 12% O2+5% CO2 were breathed, but TGF beta1 did not change. A significant linear relationship of VEGF and bFGF mRNA to minute ventilation was observed in awake animals (r=0.98, p<0.02 and r=0.87, p<0.03, respectively). The paralysed, mechanically-ventilated animals showed no mRNA increases for any probe. Systemic hypoxia had no additional effect on VEGF or bFGF levels in the diaphragm. It was concluded that messenger ribonucleic acid for vascular endothelial growth factor and basic fibroblast growth factor in the diaphragm rises significantly as a result of active ventilation and not due to blood gas/pH changes or to passive muscle shortening per se. PMID- 11401065 TI - Whistle mouth pressure as test of expiratory muscle strength. AB - Expiratory muscle strength is a determinant of cough function. Mouth pressures during a maximal static expiratory effort (PE,max) are dependent on patient motivation and technique and low values are therefore difficult to interpret. This study hypothesized that a short, sharp and maximal expiration through a narrow aperture, a "whistle", might provide a complementary test of expiratory muscle strength. To obtain a maximal whistle, subjects (27 healthy volunteers and 10 patients with amyotrophic lateral sclerosis) were asked to perform a short, sharp blow as hard as possible, from total lung capacity, through a reversed paediatric inhaler whistle, connected to a flange-type mouthpiece. In both healthy subjects and patients, whistle mouth pressure (Pmo,W) was closely related to the pressure measured in the oesophagus and stomach during the same manoeuvre. In healthy subjects, Pmo,W and PE,max correlated with wide limits of agreement, although Pmo,W values were significantly higher than PE,max (131+/-31 cmH2O versus 101+/-27 cmH2O, p<0.0001). In patients, it was also found that Pmo,w and PE,max values were strongly related (r=0.937, p<0.0001). In healthy subjects, the intraclass correlation coefficient and the variation coefficient for Pmo,W repeated measurements were respectively 0.88 and 7.0%. However Pmo,W and PE,max were always smaller than the gastric pressure generated by a maximal cough. It is concluded that mouth whistle pressure, a noninvasive, reproducible and simple test, provides a reliable measure of expiratory muscle strength in healthy subjects that is acceptable to patients and can be used in a complementary fashion to maximal static expiratory effort. PMID- 11401066 TI - Familial aggregation and heritability of adult lung function: results from the Busselton Health Study. AB - Decreased spirometric indices are characteristic of asthma and other respiratory diseases. The aim of this study was to investigate the genetic and environmental components of variance of forced expiratory volume in one second (FEV1) and forced vital capacity (FVC) measured in adulthood in an Australian population based sample of 468 Caucasian nuclear families. The inter-relationships of the genetic determinants of these traits with asthma and atopic rhinitis were also investigated. Serial cross-sectional studies were conducted in the town of Busselton in Western Australia between 1966 and 1981 and follow-up of previous attendees was undertaken in 1995. Data from each subject included in this study were from a single survey in adulthood (25-60 yrs of age) when the subject was as close to age 45 yrs as possible. Multivariate analysis suggested that FEV1 and FVC levels were associated with age, sex, height, tobacco smoke exposure, asthma and atopic rhinitis. After adjustment for relevant covariates, FEV1 levels had a narrow-sense heritability (h2N) of 38.9% (SE 9.1%). FVC levels had an h2N of 40.6% (SE 8.9%). Extended modelling demonstrated little overlap in the genetic determinants of asthma or atopic rhinitis and either FEV1 or FVC levels. The results of this study were consistent with the existence of important genetic determinants of adult lung function that are independent of asthma or other atopic disease, cigarette smoking, height, age or sex. PMID- 11401067 TI - Elastic properties of the lung and the chest wall in young and adult healthy pigs. AB - Understanding of the elastic pressure/volume (Pel/V) curve is still limited in health and disease. The aim of the present study was to elucidate the Pel/V curve and elastance of the respiratory system (ERS) lung (EL) and chest wall (ECW) in healthy pigs. Six young (20.8 kg) and seven adult (58.9 kg), anaesthetized, paralysed and ventilated pigs were studied. Pel/V curves were recorded at zero end-expiratory pressure (ZEEP) and at positive end-expiratory pressure (PEEP) up to 40 cmH2O with a computer controlled ventilator during an insufflation at a low, constant flow. Pel/V curves of the respiratory system showed a complex pattern in both young and adult pigs. During the insufflation, ERS decreased, increased, fell, and increased again. A second Pel/V curve recorded immediately after the first one showed lower elastance and only one early fall in ERS. ECW fell over the initial segment and was then nearly stable. Difference between 1st and 2nd curves reflected changes in EL caused by recruitment during the 1st insufflation. At PEEP, such signs of collapse and recruitment were reduced. A strong tendency to lung collapse contributes to a complex pattern of elastic pressure/volume curves. At low volumes and distending pressures the chest wall contributes significantly to changes in respiratory system elastance. PMID- 11401068 TI - Pulmonary function, inflammation, exercise capacity and quality of life in cystic fibrosis. AB - The aim of the study was to determine the extent to which treatment induced changes in exercise capacity and quality of life (QoL) are related to spirometric measures of lung function and other measures of disease impairment. Twenty patients admitted to hospital with an exacerbation of pulmonary disease were recruited. Measures of disease impairment, disability and QoL were obtained at the beginning and end of an intravenous course of antibiotic therapy. Intravenous antibiotic treatment resulted in a significant improvement in all measures of disease impairment, disability and handicap. The only significant predictor of treatment induced change in exercise capacity was C-reactive protein (CRP) and this explained 28% of the variance in change in exercise capacity. In the case of QoL, two predictors (change in exercise capacity and sputum output) contributed significantly to the change in QoL and collectively explained 54% of the variance in QoL. Lung function provides a limited index of treatment outcome. Exercise capacity and quality of life assessment have the potential to make a significant contribution to the decision making process regarding treatment choices in cystic fibrosis and should be measured directly if a comprehensive evaluation of the effect of treatment is required. PMID- 11401069 TI - Surfactant protein A and other bronchoalveolar lavage fluid proteins are altered in cystic fibrosis. AB - Inflammation and proteolytic processes play an important role in the progression of cystic fibrosis (CF) lung disease. The goal of this study was to describe bronchoalveolar lavage fluid (BALF) protein pattern of CF patients in comparison to controls and to assess if there is proteolytic degradation of surfactant protein A (SP-A), an important innate host defence component of the lungs. BALFs from 17 clinically stable CF patients and from eight healthy children were separated by two-dimensional gel electrophoresis. Silver staining was used to show BALF proteins and Western blotting to detect SP-A isoforms. In CF, BALF proteins of a low molecular weight < or = 20 kD were more abundant than in controls. Various proteins were seen in CF which were not present in controls and vice versa. Degradation of SP-A was present in 15 of 17 CF BALFs but in none of the controls, in contrast polymeric isoforms were seen in all controls and in four of 17 CF patients. Proteolytic damage to surfactant protein A and significant changes of normal bronchoalveolar lavage fluid proteins occur in lungs of cystic fibrosis patients. Identification of altered bronchoalveolar lavage fluid proteins may give new insights into pathogenic mechanisms and provide new targets for therapy. PMID- 11401070 TI - Sleep fragmentation: comparison of two definitions of short arousals during sleep in OSAS patients. AB - The measurement of arousals during sleep is useful to quantify sleep fragmentation. The criteria for electroencephalography (EEG) arousals defined by the American Sleep Disorders Association (ASDA) have recently been criticized because of lack of interobserver agreement. The authors have adopted a scoring method that associates the increase in chin electromyography (EMG) with the occurrence of an alpha-rhythm in all sleep stages (Universite Catholique de Louvain (UCL) definition of arousals). The aim of the present study was to compare the two scoring definitions in terms of agreement and repeatability and the time taken for scoring in patients with obstructive sleep apnoea syndrome (OSAS) of varying severity. Two readers using both ASDA and UCL definitions scored twenty polysomnographies (PSGs) each on two occasions. The PSGs were chosen retrospectively to represent a wide range of arousal index (from 6-82) in OSAS patients. There was no difference in the arousal indices between readers and between scoring methods. The mean+/-SD difference between the two definitions (the bias) was 1.1+/-3.76 (95% confidence interval: -0.66-2.86). There was a strong linear relationship between the arousal index scored with the two definitions (r=0.981, p<0.001). Mean+/-SD scoring duration was significantly shorter for UCL than for ASDA definitions (18.5+/-5.4 versus 25.3+/-6.6 min, p<0.001). In conclusion, it has been found that in obstructive sleep apnoea syndrome patients, the American Sleep Disorders Association and Universite Catholique de Louvain definitions were comparable in terms of agreement and repeatability. PMID- 11401071 TI - Angiotensin converting enzyme in patients with sleep apnoea syndrome: plasma activity and gene polymorphisms. AB - The prevalence of several cardiovascular diseases is increased with obstructive sleep apnoea syndrome (OSAS), due to, as yet, unclear reasons. Angiotensin converting enzyme (ACE) abnormalities have been implicated in the pathogenesis of various cardiovascular diseases. In this study, plasma ACE activity and the distribution of an insertion (I)/deletion (D) polymorphism of the ACE gene were determined in OSAS patients and in healthy controls. A total of 63 patients with OSAS (mean+/-SEM 54.5+/-2.5 apnoea/hypopnoeas.h(-1)) and 32 healthy subjects were studied. To avoid potential confounding factors, patients treated with ACE inhibitors or continuous positive airway pressure were excluded, as well as controls in whom a blood sample was not obtained early in the morning. ACE activity was determined spectrophotometrically in 46 OSAS patients and 25 controls. The I/D ACE polymorphism was determined by polymerase chain reaction in 44 patients and 32 controls. ACE activity was higher in OSAS patients (53.9+/-2.5 IU.L(-1)) than in healthy controls (42.4+/-3.1 IU.L(-1), p<0.01). This was independent of the presence of arterial hypertension. The frequency distribution of the DD, II and ID genotypes in OSAS patients (30%, 16%, 54%, respectively) was not significantly different from that seen in healthy subjects (31%, 28%, 41%, respectively, p=0.356). These results indicate that ACE plasma activity is increased in untreated OSAS patients. This increased activity may contribute to the pathogenesis of the cardiovascular disease in these patients. PMID- 11401072 TI - Health effects of diesel exhaust emissions. AB - Epidemiological studies have demonstrated an association between different levels of air pollution and various health outcomes including mortality, exacerbation of asthma, chronic bronchitis, respiratory tract infections, ischaemic heart disease and stroke. Of the motor vehicle generated air pollutants, diesel exhaust particles account for a highly significant percentage of the particles emitted in many towns and cities. This review is therefore focused on the health effects of diesel exhaust, and especially the particular matter components. Acute effects of diesel exhaust exposure include irritation of the nose and eyes, lung function changes, respiratory changes, headache, fatigue and nausea. Chronic exposures are associated with cough, sputum production and lung function decrements. In addition to symptoms, exposure studies in healthy humans have documented a number of profound inflammatory changes in the airways, notably, before changes in pulmonary function can be detected. It is likely that such effects may be even more detrimental in asthmatics and other subjects with compromised pulmonary function. There are also observations supporting the hypothesis that diesel exhaust is one important factor contributing to the allergy pandemic. For example, in many experimental systems, diesel exhaust particles can be shown to act as adjuvants to allergen and hence increase the sensitization response. Much of the research on adverse effects of diesel exhaust, both in vivo and in vitro, has however been conducted in animals. Questions remain concerning the relevance of exposure levels and whether findings in such models can be extrapolated into humans. It is therefore imperative to further assess acute and chronic effects of diesel exhaust in mechanistic studies with careful consideration of exposure levels. Whenever possible and ethically justified, studies should be carried out in humans. PMID- 11401073 TI - Respiratory symptoms in European animal farmers. AB - Farmers are known to be at high risk for the development of occupational airway disease. The aim of this European study was to determine which airway symptoms predominate in different types of animal farmers (cattle, pigs, poultry, sheep) and to compare the prevalence of symptoms to the general population. A total of 6,156 randomly selected animal farmers in Denmark, Germany (Schleswig-Holstein, Niedersachsen), Switzerland, and Spain completed a questionnaire on respiratory symptoms and farming characteristics in 1995-1997. The prevalence of general respiratory symptoms was compared to the results of the European Community Respiratory Health Survey (ECRHS) obtained in the same regions. Pig farmers were at highest risk for the development of work-related symptoms. A significant dose response relationship between daily hours worked inside animal houses and symptoms was established for pig and poultry farmers. Additionally, self-reported nasal allergies (odds ratio (95% confidence interval): 3.92 (3.26-4.71)) and nasal irritation during work (3.98 (3.35-4.73)) were shown to be associated with the development of chronic phlegm. The prevalence of wheezing, shortness of breath, asthma and nasal allergies was signficantly lower among all farmers in the age group 20-44 yrs than among the general population. However, the prevalence of usually bringing up phlegm in winter among farmers was significantly higher than in the general population (9.4 (8.3-10.5%) versus 7.5 (6.5-8.5%)). Individual factors have been shown to be related to the prevalence of chronic phlegm among farmers. Additionally, this study could support the hypothesis that farming could be negatively related to allergic diseases. PMID- 11401074 TI - The infant lung function model: a mechanical analogue to test infant lung function equipment. AB - To facilitate international multicentre studies and quality control of infant pulmonary function measurements, the European Respiratory Society-American Thoracic Society (ERS-ATS) working group for infant lung function testing aims to develop specifications for standardized infant lung function equipment and software. However, a standardized test device is also needed to test whether existing infant lung function equipment is able to meet these requirements. The authors have built a "mechanical model baby" consisting of a linear pump which can reproduce prerecorded tidal flow waveforms with a precision of 0.5% (full stroke), enabling the simulation of tidal and forced flow patterns. This linear pump can be connected to a series of copper lung volumes (range 50-300 mL) with known time constants, so that lung volumes can be reproduced with a precision of +/-1% at frequencies 10-120bpm. Five airflow resistors were built using sinter material. When assessed using flows 0-300 mL.s(-1) all resistors showed a quasilinear pressure/ flow relationship, with slopes 1.0-5.6 kPa.L(-1).s. These resistances could be reproduced with a precision of +/-2.5%. The infant lung model can also be used to assess frequency responses of infant lung function equipment, since the pump is capable of delivering low amplitude volumes up to 20 Hz in a pseudorandom noise manner. In summary, based on error estimations, this infant lung model is able to test whether or not infant lung function equipment meets the requirements suggested by the European Respiratory Society-American Thoracic Society standardization group, that is: flow measurements within +/ 2.5%, volume and resistance measurements within +/-5%, frequency response: magnitude attenuation <+/-10% and phase shift <+/-3 degrees at 10 Hz. PMID- 11401075 TI - Assessment of an infant whole-body plethysmograph using an infant lung function model. AB - In order to facilitate international multicentre studies and improve the quality control of infant pulmonary function measurements, the European Respiratory Society-American Thoracic Society Task Force for infant lung function testing has recently developed specifications for standardized infant lung function equipment and software. A mechanical infant lung model analogue has been developed to assess whether infant lung function equipment is able to meet these requirements. However, the practical testing of infant lung function equipment using such models is highly complex because of the need to use very small pressure and flow changes, and the numerous potentially confounding factors associated with both the design of the device and the testing procedure. The aim of this study was to determine whether the infant lung model is capable of assessing the overall function of an whole-body infant- plethysmograph, using the only infant plethysmograph that was commercially available at the time as an example. The mechanical characteristics of the model such as vibrations or noise did not disturb the delicate plethysmographic measurements and thereby allowed a reliable assessment of the system. A series of tests revealed that the plethysmograph was able to measure airway resistance 1-3.5 kPa.L(-1).s with an accuracy of +/-2.5% and lung volumes 75-300 mL with an accuracy of +/-2.5% under in vitro conditions. To conclude, the infant lung model is a useful means of assessing the overall in vitro performance of infant whole-body plethysmographs, but thermal, mechanical and frequency response characteristics of such a device must be taken into account when interpreting the results of such assessments. PMID- 11401076 TI - Allergy markers in respiratory epidemiology. AB - Assessing allergy by measurement of serum immunoglobulin (Ig) E antibodies is fast and safe to perform. Serum antibodies can preferably be assessed in patients with dermatitis and in those who regularly use antihistamines and other pharmacological agents that reduce skin sensitivity. Skin tests represent the easiest tool to obtain quick and reliable information for the diagnosis of respiratory allergic diseases. It is the technique more widely used, specific and reasonably sensitive for most applications as a marker of atopy. Measurement of serum IgE antibodies and skin-prick testing may give complimentary information and can be applied in clinical and epidemiological settings. Peripheral blood eosinophilia is less used, but is important in clinical practice to demonstrate the allergic aetiology of disease, to monitor its clinical course and to address the choice of therapy. In epidemiology, hypereosinophilia seems to reflect an inflammatory reaction in the airways, which may be linked to obstructive airflow limitation. PMID- 11401077 TI - Microbial investigation in ventilator-associated pneumonia. AB - Ventilator-associated pneumonia (VAP) is a serious infectious condition in intensive care unit (ICU) patients, currently related to a high mortality rate. Therefore, this complication of mechanical ventilation requires a prompt diagnosis and adequate antibiotic treatment. The detection of the causative organism is imperative for guiding an appropriate therapy as there is strong evidence of the adverse effect of inadequate empirical treatment on outcome. The major difficulty of the microbial investigation is to obtain the sample from the lower respiratory tract, mainly because of the potential contamination with upper airways flora, which may result in a misinterpretation of the cultures. Microbial investigation in VAP is based on the culture of samples obtained from lower respiratory tract by noninvasive or invasive methods. The most common techniques of sampling are the endotracheal aspirate (ETA), which is considered a noninvasive method, the protected specimen brush (PSB) and the bronchoalveolar lavage (BAL), both being invasive methods of investigation. The latter were designed as an attempt to avoid the colonizing flora of the upper airways. The best of these diagnostic approaches is still controversial. In terms of outcome, there is strong evidence that the impact of both invasive and noninvasive methods seems to be similar. In terms of cost, however, the endotracheal aspirate is less expensive compared to BAL or PSB. On the other hand, invasive methods could be particularly beneficial in patients who are not responding to the initial empirical antibiotic treatment. The rationale for the quantitative culture of the respiratory samples is to differentiate between infection and colonization of lower airways, because the bacterial colonization is a frequent event in mechanically ventilated patients. The thresholds currently employed for the diagnosis of the pneumonia are the following: ETA samples, > or = 10(5)-10(6) colony forming units (cfu).mL(-1); PSB samples, > or =10(3) cfu.mL(-1); and BAL samples, > or =10(4) cfu.mL(-1). Intending to provide a practical approach to the issue, the present manuscript reviews the available noninvasive (blood culture, endotracheal aspirate) and invasive (protected specimen brush, bronchoalveolar lavage, blinded methods and lung biopsy) techniques used for the diagnosis of ventilator-associated pneumonia. PMID- 11401078 TI - The role of positron emission tomography with 18F-fluoro-2-deoxy-D-glucose in respiratory oncology. AB - In the past 5 yrs, positron emission tomography (PET) with 18F-fluoro-2-deoxy-D glucose (FDG) has become an important imaging modality in lung cancer patients. At this time, the indication of FDG-PET as a complimentary tool to computed tomography in the diagnosis and staging of nonsmall cell lung cancer has gradually gained more widespread acceptance and also reimbursement in many European countries. This review focuses on the data of FDG-PET in the diagnosis of lung nodules and masses, and in locoregional and extrathoracic staging of nonsmall cell lung cancer. Emphasis is put on the potential clinical implementation of the currently available FDG-PET data. The use of FDG-PET in these indications now needs further validation in large-scale multicentre randomized studies, focusing mainly on treatment outcome parameters, survival and cost-efficacy. Interesting findings with 18F-fluoro-2-deoxy-D-glucose-positron emission tomography have also been reported for the evaluation of response to radio- or chemotherapy, in radiotherapy planning, recurrence detection and assessment of prognosis. Finally, a whole new field of application of positron emission tomography in molecular biology, using new radiopharmaceuticals, is under extensive investigation. PMID- 11401079 TI - The use of positive pressure devices by physiotherapists. AB - The aim of this paper is to review the indications for use by physiotherapists, such as physiological rationale and the comparative efficacy of intermittent positive pressure breathing (IPPB) and continuous positive airway pressure (CPAP). A brief discussion of nasal intermittent positive airway pressure is also included. The use of IPPB for post operative prophylaxis has not been supported in the literature. In patients with low lung volumes resulting from neuromuscular disease or spinal injury, IPPB may be useful in the acute phase to improve tidal volume and cough effectiveness. The physiological benefits of CPAP to improve lung volumes are well documented in the literature. Physiotherapists use CPAP as an intermittent application in patients with low lung volumes following surgery. It is predominantly used as a second line intervention in the presence of refractory atelectasis and poor gas exchange. It may also be indicated in other patient groups with similar physiological problems. Nasal intermittent positive airway pressure combines the beneficial effects of intermittent positive pressure breathing and continuous positive airway pressure. There have been many studies evaluating its effectiveness. These have been supportive for patients with neuromuscular disease and sleep disordered breathing, but more research is needed in patients with acute respiratory failure. PMID- 11401080 TI - Successful treatment of severe respiratory papillomatosis with intravenous cidofovir and interferon alpha-2b. AB - In contrast to uncomplicated juvenile laryngeal papillomas which may regress spontaneously, papillomatosis involving pulmonary parenchyma is associated with a poor outcome. This report represents the case of a 34 yr-old female in whom respiratory papillomatosis resulted in voice problems and recurrent pneumonias due to bronchial obstruction. A computed tomographic scan of the chest showed worsening bilateral round cavitating consolidations. Bronchoscopy revealed polypoid lesions of the right vocal cord and the midtrachea which were confirmed as squamous papillomatosis by histology. Interferon (IFN)alpha-2b treatment was inefficient as was cidofovir monotherapy on a maintenance basis. Six months of IFN-alpha-2b and cidofovir in combination led to a complete macroscopic disappearance of the laryngeal lesions and to an impressive regression of the tracheal papillomas and of the intrapulmonary consolidations. These data provide support that severe respiratory papillomatosis can be safely treated by interferon alpha-2b and cidofovir in combination. Probably the same mechanisms as in ribavirin plus interferon alpha-2b, in the treatment of patients with chronic hepatitis C, are responsible for the therapeutic success in this case. PMID- 11401081 TI - Sputum induction in young cystic fibrosis patients. PMID- 11401082 TI - Course and prognosis of sarcoidosis in African-Americans versus Caucasians. PMID- 11401083 TI - Thrombin converts singlet oxygen (1O2)-oxidized fibrinogen into a soluble t-PA cofactor. A new method for preparing a stimulator for functional t-PA assays. AB - Activated phagocytes, particularly polymorphonuclear leukocytes (neutrophils), by means of oxidative photonic burst, i.e., the combined activation of NADPH-oxidase and myeloperoxidase, generate large amounts of oxidants of the hypochlorite/chloramine type that are an important physiologic source for the nonradical, photon-emitting oxidant singlet oxygen (1O2), which (in the dark blood stream) is both a signal and an agent of defense against bacteria or fibrin. 1O2-oxidized fibrinogen or oxidized fibrin monomer has previously been shown to be unpolymerizable, and methionine to methionine sulfoxide-oxidized fibrinogen occurs in circulating blood. The present study demonstrates that thrombin converts oxidized fibrinogen into a soluble stimulator of tissue-type plasminogen activator (t-PA). After addition of 0.1 IU thrombin to 25 microl oxidized normal human plasma and an incubation time of 10 min (room temperature), t-PA activity increases about 20-fold when compared with oxidized plasma without the addition of thrombin. Thus, since oxidized fibrin monomer is a t-PA cofactor, thrombin-degraded oxidized fibrinogen can be used as a stimulator in functional t PA assays. PMID- 11401084 TI - Anemia of chronic disease is the more frequent type of anemia seen in patients with chronic idiopathic neutropenia of adults. AB - This study describes the frequency and the type of anemia seen in patients with nonimmune chronic idiopathic neutropenia of adults (NI-CINA). We found that NI CINA patients had low hemoglobin levels and increased serum concentrations of erythropoietin (EPO), tumor necrosis factor-alpha (TNF-alpha), and interleukin 1beta (IL-1beta). The hemoglobin levels correlated positively with the number of circulating neutrophils and inversely with the levels of EPO and TNF-alpha but not of IL-1beta. Anemia, defined as the reduction of the hemoglobin below 12.0 g/dl for women and 13.3 g/dl for men, was found in 23 out of 148 patients studied, a proportion of 15.5%. Two of the anemic patients had iron deficiency anemia (8.7%), 11 had anemia of chronic disease (ACD; 47.8%) presenting with normal or slightly reduced erythrocytic indices, low serum iron, and increased serum ferritin, and the remaining ten had anemia of undefined pathogenesis (AUP; 43.5%) with normal or slightly decreased erythrocytic indices, serum iron ranging from 43 to 88 microg/dl, and ferritin values ranging from 12 to 50 ng/ml. We conclude that ACD is the more frequent type of anemia seen in patients with NI CINA, and that pro-inflammatory cytokines, notably TNF-alpha, may be involved in the pathogenesis of both ACD and AUP, given that serum levels of the cytokine were significantly increased and that the EPO response to anemia was blunted in these patients. These findings further support our previously reported suggestion for the possible existence, in NI-CINA patients, of an unrecognized low-grade chronic inflammatory process that may be involved in the pathogenesis of the disorder. PMID- 11401085 TI - Analysis of stem cell apheresis products using intermediate-dose filgrastim plus large volume apheresis for allogeneic transplantation. AB - Previously, a dose-dependent influence of recombinant human granulocyte colony stimulating factor (rhG-CSF) on CD34+ mobilization was demonstrated. In this single-center prospective analysis, 52 healthy donors were investigated to determine the efficacy of intermediate-dose rhG-CSF 2x8 microg/kg donor body weight (bw) and intermediate large volume apheresis (LVA, median 12 l) to mobilize peripheral blood progenitor cells (PBPC) for allogeneic transplantation. The median number of CD34+ cells in apheresis products was 0.45% and 2.2x10(6)/kg recipient bw per single apheresis. A total of 5.4x10(6)/kg CD34+ cells were collected with two (range: one to three) LVA. In the analysis of donor subgroups, higher peripheral blood (PB) and apheresis results were obtained in male vs female donors; however, donor weight significantly differed in both groups. Heavier donors displayed higher PB and apheresis CD34+ counts; however, when CD34+ cells/kg were adjusted to a constant bw, similar harvest results were calculated in males and females, demonstrating that gender per se does not, whereas bw does affect apheresis results. Younger donors had significantly higher PB CD34+ counts, higher CD34+ numbers per single apheresis, increased CFU, more T, B, and CD61+, comparable NK, and less CD14+ cells. A correlation analysis of donor age and apheresis results displayed an age-related decline of 0.46x10(6)/kg CD34 cells per decade of donor aging. Cell subsets in apheresis products were CD14 (49%), CD3 (22%), CD4 (13%), CD8 (7%), CD61 (20%), CD19 (5%), and CD16/56+ (3%) cells, with increasing CD14+ cells and decreasing CD3, CD4, CD8, CD61, CD19, and CD16/56+ cells on subsequent days of apheresis. Compared to our previous analysis using high- (2x12 microg) and low-dose (1x10 microg) rhG-CSF for allogeneic PBPC mobilization, the intermediate-dose showed a similar CD34+ mobilization potential to 1x10 microg rhG-CSF; however, with use of LVA, two instead of three (p<0.05) aphereses were sufficient to mobilize > or =4x10(6)/kg bw CD34+ cells in most donors. Taken together, our results demonstrate that intermediate-dose rhG-CSF sufficiently mobilizes > or =4x10(6)/kg x bw CD34+ cells with use of LVA and that especially younger donors display increased CD34+ cell numbers. PMID- 11401086 TI - Anti-thymocyte-globulin as part of the preparative regimen prevents graft failure and severe graft versus host disease (GvHD) in allogeneic stem cell transplantation from unrelated donors. AB - To reduce the incidence of GvHD and the rate of graft failure in unrelated stem cell transplantation, we incorporate anti-thymocyte globulin in the preparative regimen in 98 patients with hematological or inherited storage disease. The median age was 32 years (range: 1-56) and 84 patients underwent transplantation from HLA-A,-B and DR identical donor, while in 14 patients the donor were mismatched either in HLA- A, -B or -DR locus. Only one patient with chronic myelocytic leukemia (CML) and blast crisis had a primary graft failure (1%). Grade II-IV acute GvHD occurred in 37 patients (37%), grade III/IV GvHD developed in 15 patients (15%). Chronic GvHD was observed in 29%, and only 12 patients had extensive GvHD (17%). After a median follow-up of 34 months (range, 9-90), the estimated overall survival at 3 years for all patients is 58% (CI 95%: 48%-68%), and the estimated disease-free survival at 3 years is 49% (CI 95%: 38%-60%). For patients with CML transplanted in first chronic phase or accelerated phase (n=40), the estimated overall survival at 3 years is 70% (CI 95%: 56%-84%), and the estimated disease-free survival at 3 years is 58% (CI 95%: 17%-85%). ATG in unrelated stem cell transplantation reduces the risk of severe acute and chronic GvHD and of graft failure without an obvious increase of severe infection. Further follow-up is mandatory to determine the incidence of late relapse. PMID- 11401087 TI - Clinical and immunological evaluation of primary splenic irradiation in chronic lymphocytic leukemia: a study of 24 cases. AB - Little is known about the effects of primary splenic irradiation (SI) in B-cell chronic lymphocytic leukemia (B-CLL) on subpopulations of lymphocytes, immunoglobulins, and the induction of autoantibodies against erythrocytes and platelets. Twenty-four untreated patients with B-CLL were studied prospectively. One patient was excluded from analysis because of intercurrent death. Of the remaining 23 patients, 7 were female and 16 were male, with an average age of 68.4 and 61.2 years, respectively. Treatment consisted of a weekly dose of I Gy up to a total dose of 10 Gy to the spleen. Standardized evaluation was done 2 weeks before and 6 weeks after SI. Twenty patients completed the course of SI. In 14 patients there was a partial response and in 9 patients the disease remained stable. The number of leukocytes decreased rapidly and significantly, with a decrease in the fraction of lymphocytes and an increase in the neutrophil count (all P<0.0001). There was a significant increase in platelet count, but not in hemoglobin level. Nausea, slight diarrhea, pleuropneumonia, granulopenia with fever (all n=1), urinary tract infection and high fever (n=1), and thrombocytopenia occurred (n=2). One patient received transfusion of packed cells because of upper digestive tract bleeding for which cessation of SI was not necessary. The number of malignant cells showed a significant decrease (P<0.01). No significant changes in CD4/CD8 ratio were found. The number of CD4+ and CD8+ cells, however, decreased significantly (P<0.01 and 0.03, respectively). DAT remained unchanged in most (93%) and tests for antibodies to platelets in 83% of the patients. No significant changes in immunoglobin levels were observed. PMID- 11401088 TI - Intravascular B-cell lymphoma in a 38-year-old woman: a case report. AB - Intravascular lymphoma (IVL) is a rare aggressive disease characterised by the presence of lymphoma cells only in the lumina of small vessels, particularly capillaries. Only about 200 cases have been reported in the world (some of them retrospectively). IVL is predominantly of B-cell lineage origin but occasionally T-cell lineage occurs. Multiple organs may be involved and a variety of clinical presentations have been described. These include nephrotic syndrome, pyrexia and hypertension, breathlessness and haemolytic anaemia, leukopoenia, pancytopoenia and disseminated intravascular coagulation. We report a case of a 38-year-old woman with a highly aggressive clinical course of IVL. She was admitted to the Department of Neurosurgery because of spondylolisthesis of L5-S1 qualified to surgery. During hospitalisation haemolytic anaemia, thrombocytopoenia and splenomegaly were observed and she was admitted to the Department of Haematology for diagnosis. During her staying in the hospital, new symptoms, such as kidney and liver failure, occurred and the central nervous system was involved. The clinical course was very rapid and progressive. Corticosteroid therapy was started but the disease soon led to the fatal outcome. Diagnosis was established at post-mortem examination. PMID- 11401089 TI - Agranular CD4+CD56+ blastic natural killer leukemia/lymphoma. AB - Blastic natural killer cell leukemia/lymphoma (blastic NKL/L) is characterized by blastic morphology and a distinctive immunophenotype combining blastic features and cytologically resembling acute myeloid or lymphoid leukemia. The clinical, pathologic, and cytogenetic features of blastic NKL/L have not yet been systematically identified. We report herein a case of blastic NKL/L with skin lesion, adenopathy, and systemic lymphoadenopathy. The identified tumor cells were positive for CD4 and CD56, and negative for T-cell, B-cell, and myeloid markers. T-cell receptor beta, gamma, delta, and immunoglobulin heavy chain genes in the bone marrow cells showed germ-line configurations. Southern blot analysis with a terminal probe did not reveal any Epstein-Barr virus infection. Although patients diagnosed as blastic NKL/L have generally shown chemotherapy resistance and poor prognosis, our patient was treated with a combined chemotherapy, which is also used for acute lymphoblastic leukemia, and has maintained complete remission (CR) for more than 13 months. In addition to clinical investigations, we thoroughly analyzed his karyotype by using a combination of G-banding and a new technique, spectral karyotyping. The karyotype was described as 45, XY, der(1)t(1;20)(p32;q11.2), der(6) (1pter-->1p32:: 6p21.1-->6q13:: 7q11.2-->7qter), der(7) t(7;20)(q11.2;q11.2), t(13;14)(q14;q32), der(13)t(6;13) (p21.1; q14), -20. PMID- 11401090 TI - Paraneoplastic polyneuropathy preceding the diagnosis of Hodgkin's disease and non-small cell lung cancer in a patient with concomitant Borrelia burgdorferi infection. AB - A patient with painful peripheral neuropathy is presented, whose symptoms were thought to result from an infection with Borrelia burgdorferi sensu lato. Investigations of the cerebrospinal fluid for signs of inflammation and borrelial antibodies were negative, and the patient did not benefit from repeated antibiotic treatment. Electrophysiologic studies and sural nerve biopsy showed axonal neuropathy consistent with a paraneoplastic syndrome. Further workup revealed mediastinal Hodgkin's disease (HD; nodular sclerosing subtype) Ann Arbor stage II and non-small cell cancer of the lung (stage T1N0M0). Surgical resection of the lung cancer and combined chemo- and radiotherapy for HD resulted in complete remission of both malignancies. While the preexisting neurologic symptoms persisted during treatment, neurography showed some improvement of the distal nerves. During radiation therapy the patient developed transient left sided brachial plexopathy. This case illustrates that the diagnosis of borreliosis in patients with isolated painful peripheral neuritis cannot be based solely upon positive IgG titers and supports the requirement for a thorough workup for an underlying--potentially curable--disease. In addition, singular pulmonary lesions in the setting of HD should be suspected to have a separate cause. PMID- 11401091 TI - "Pseudotumor cerebri" following allogeneic bone marrow transplantation (BMT). AB - Pseudotumor cerebri, an uncommon complication following BMT, has been generally associated with cyclosporin A neurotoxicity. However, it has not previously been reported as a clinical presentation of sinusitis in spite of its high incidence after BMT. We report a case of pseudotumor cerebri secondary to sinusitis in a child with acute lymphoblastic leukemia and who underwent unrelated bone marrow transplantation. PMID- 11401092 TI - Avascular necrosis in the femoral head secondary to bone marrow infarction in a patient with graft-versus-host disease after unrelated bone marrow transplantation. AB - We previously reported a case of bone marrow infarction attributable to acute graft-versus-host disease (GVHD) in a patient with acute lymphoblastic leukemia after unrelated bone marrow transplantation (BMT). Although the bone marrow infarction-induced arthralgia in this patient improved, severe arthralgia appeared again with exacerbation of chronic GVHD, and the arthralgia was strongly correlated with the clinical course of chronic GVHD, i.e., the course of symptoms such as dermal and hepatic GVHD and ocular dryness. Finally, the patient developed avascular necrosis (AVN) in the right femoral head. Serum interleukin (IL)-6 and IL-10 levels were high at the onset of arthralgia but low during remission, and levels of interferon-gamma were undetectable throughout the period of arthralgia. Based on the clinical course and these data, chronic GVHD was thought to have been the major cause of the AVN. Since IL-10 antagonizes various other cytokines that induce GVHD, the increase in IL-10 might have inhibited the development of GVHD. PMID- 11401093 TI - Concurrent pernicious anemia and myelodysplastic syndrome. AB - Megaloblastic anemia (MA) due to vitamin B12 deficiency is a reversible form of ineffective hematopoiesis. Myelodysplastic syndrome (MDS) is an acquired, irreversible disorder of ineffective hematopoiesis, characterized by stem cell dysfunction as a consequence of DNA damage manifested in part by karyotype anomalies. Importantly, MA and MDS are generally considered mutually exclusive diagnoses. We report the case of a 73-year-old woman with a profound macrocytic anemia, monocytosis and neurologic symptoms. Low cobalamin levels and the presence of anti-intrinsic-factor antibodies definitively established a diagnosis of pernicious anemia. Replacement therapy resulted in resolution of neurologic findings and macrocytosis; however, the anemia and monocytosis persisted. Bone marrow biopsy revealed trilineage myelodysplasia, which together with the peripheral monocytosis suggested a diagnosis of chronic myelomonocytic leukemia. Karyotype analysis revealed a clone with 45, XX, +der(1;7)(q10;p10)-7 [20]. Eighteen months after documented vitamin B12 replenishment her MDS transformed to terminal acute myeloid leukemia with the same clonal abnormality. Reversible cytogenetic abnormalities have been observed with MA, occasionally including karyotypes typically associated with MDS or myeloid leukemias. These abnormalities, like the anemia, resolve with vitamin replacement. This case suggests that MA and MDS can occur simultaneously; clinicians should be aware that this phenomenon occurs. Whether acquired karyotype abnormalities from the MA were related to the MDS and subsequent myeloid leukemia in this woman is a speculative but intriguing consideration that is discussed. PMID- 11401094 TI - Acute promyelocytic leukemia with additional chromosome abnormalities in a renal transplant case. AB - Some cases of acute myeloid leukemia following organ transplant (PT-AML) have been published in the literature. We report the second case of acute promyelocytic leukemia (APL), which developed post-transplant and immunosuppressive treatment, in a 50-year-old male who had undergone a renal transplant. At diagnosis he presented typical t(15;17)(q12;q13) with additional abnormalities, including +8,t(13;22)(q12;q13) and an abnormal chromosome 1 which was better characterized by fluorescence in situ hybridization (FISH). He obtained cytological, karyotypic and molecular complete remission (CR) with induction treatment according to the all-trans retinoic acid + idarubican (AIDA) protocol; after 12 months, he relapsed (molecular relapse) and achieved molecular remission with all-trans retinoic acid (ATRA) plus mitoxantrone and cytosine arabinoside. After a further 14 months, he was treated with arsenic trioxide for cytological relapse and obtained a third CR; at the cytological relapse the karyotype showed 47,XY,+8, t(15;17)(q22;q21),t(13;22)(q12;q13),der(22)t(1;22)(p22;q13). He is alive 3.3 years after diagnosis of APL. Cyclosporin A (CsA) was given during all cycles of chemotherapy. We did not observe any severe infections or kidney failure during treatments. The use of conventional cytogenetic analysis plus FISH may identify complex karyotype also in transplanted patients receiving immunotherapy, and may also contribute to a better assessment of PT-AL. PMID- 11401095 TI - Antiangiogenesis in hematologic diseases. PMID- 11401096 TI - Hydrate or dehydrate. PMID- 11401097 TI - Palliative care at the National Cancer Institute of Milan. PMID- 11401098 TI - Patient education in pain control. AB - Patients' concerns about reporting pain and using analgesics have been cited as major contributors to the problem of inadequate pain management. The purpose of this paper is to describe a program of research in which we have focused on these patient concerns, or as we refer to them, "patient-related barriers to pain management." We present a summary of descriptive studies on this topic, describe educational interventions that have been tested, and suggest future avenues of research and practice. PMID- 11401099 TI - Music therapy in palliative medicine. AB - A partnership between The Cleveland Clinic Foundation and The Cleveland Music School Settlement has resulted in music therapy becoming a standard part of the care in our palliative medicine inpatient unit. This paper describes a music therapy program and its impact on patients, their families, and staff. A service delivery model is suggested for implementation and integration of music therapy within palliative medicine. Specific music therapy interventions, evaluation and documentation techniques are also mentioned. A description of patient and family responses to music therapy, staff satisfaction, and effectiveness of interventions is presented. PMID- 11401100 TI - Multidisciplinary symptom control clinic in a cancer center: a retrospective study. AB - Our objective in this study was to review the characteristics, symptom intensity and satisfaction of patients referred to a half-day symptom control clinic (SCC) for advanced cancer patients. This was a retrospective study. The setting was a multidisciplinary symptom control clinic in a cancer centre. Those taking part were 166 consecutive advanced cancer patients referred to the half-day multidisciplinary SCC because of symptom distress. Patients referred to the clinic were assessed in a private room by a physician, a nurse, a pharmacist, a psychologist, and social, rehabilitation, nutrition, respiratory and pastoral care workers. Symptom distress (multiple visual analogue scales), cognition, and CAGE (alcoholism) were determined. Recommendations were given to the patient and sent to the oncologist, family physician and home care nurse. For 110 patients a second assessment was carried out 1 week later, and 64 patients underwent a telephone assessment 2 weeks after the second visit. Symptom intensity was determined during initial and follow-up visits, as well as during two follow-up telephone assessments. In addition, demographics and patient satisfaction with the SCC were determined. Overall symptom distress, depression, anxiety and sensation of wellbeing improved significantly from the first (n = 166) to the second clinic visit (n = 110). Further significant improvement was observed in overall symptom distress, pain, anxiety, sense of wellbeing and depression at the 2- (n = 64) and 4-week (n = 38) telephone follow-up assessments. Mean satisfaction with the SCC (0-10) was 7.7 +/- 2. Our findings suggest that the work of the SCC results in long-term effectiveness in symptom control and high levels of patient satisfaction. The SCC allows for better integration of care between a cancer center and community-based physicians and nurses. It also allows patients access to multiple disciplines that are not available outside tertiary centers. PMID- 11401101 TI - The impact of pamidronate on inpatient and outpatient services among metastatic breast cancer patients. AB - Our goal was to evaluate the impact of pamidronate therapy on medical resource utilization for treatment of bone metastases among patients with breast cancer. In this 12-center retrospective study, inpatient and outpatient resource utilization was abstracted from the medical charts of 295 patients with breast cancer who were diagnosed with bone metastases between July 1996 and April 1999. Data were abstracted from the time of bone metastasis diagnosis (baseline) to the present. The analysis compared non-pamidronate patients against pamidronate patients, who were stratified on the basis of whether their pamidronate therapy had been initiated within 3 months (early pamidronate group) or more than 3 months (late pamidronate group) after diagnosis. Resource utilization was compared among groups using multivariate regression analyses. A total of 101 early pamidronate, 72 late pamidronate, and 122 non-pamidronate patients were included in the analysis. The results showed that the early pamidronate group was roughly one-half as likely to have unplanned office visits attributable to bone metastases as the late pamidronate and non-pamidronate groups. The groups had a similar likelihood of ever being hospitalized for bone-related conditions; however, among those hospitalized, there were roughly one-half as many bone related hospitalizations in the late pamidronate group as in the non-pamidronate group. Also, the mean length of stay was approximately 50% shorter in both pamidronate groups than in the non-pamidronate group. We conclude that pamidronate therapy may be associated with less medical resource utilization, particularly among patients hospitalized for bone-related conditions. PMID- 11401102 TI - Determinants of the sensation of thirst in terminally ill cancer patients. AB - While a sensation of thirst causes severe distress for a certain proportion of cancer patients in the terminal stage, the factors contributing to this symptom have not been established. To clarify the association between sensation of thirst and medical factors, especially dehydration, a cross-sectional observational study was performed on terminally ill cancer patients receiving inpatient hospice care. On admission to a palliative care unit, 88 consecutive patients underwent blood sampling and were requested to rate the intensity of thirst on a visual analogue scale (VAS). Physicians prospectively evaluated factors that might potentially be contributing to the symptom. The mean VAS score for thirst was 5.0+/-2.8, and 18% of the patients complained of severe thirst with a VAS score of > or = 8. No significant correlations were observed between the VAS score for thirst and the values of total protein, blood urea nitrogen (BUN), creatinine, sodium, osmolality, hematocrit, atrial natriuretic peptide (ANP), and biochemical dehydration defined by the levels of BUN, creatinine, sodium and osmolality. On the other hand, dehydration defined by ANP level (< or = 15 pg/ml), hyperosmolality (> or = 300 mosmol/kg), gastrointestinal cancer, survival, performance status, oral intake, vomiting, and stomatitis were significantly associated with the severity of thirst. In addition, mouth breathing and opioids were determined to be a potential clinical cause of severe thirst when a retrospective chart review was carried out. In conclusion, sensation of thirst is a frequent symptom in terminally ill cancer patients and is associated with dehydration, hyperosmolality, poor general conditions, stomatitis, oral breathing, and opioids. Careful assessments and treatment of underlying causes is important to alleviate patients' distress. PMID- 11401103 TI - Reliability and responsiveness of a prostate cancer questionnaire for radiotherapy-induced side effects. AB - Few self-assessment cancer-specific questionnaires/modules have yet been developed for radiotherapy-induced side effects. The aim of the present study was to test the reliability and responsiveness of a prostate cancer (PC)-specific questionnaire. Thirty-one patients with PC graded their urinary and intestinal symptoms and their sexual function on the questionnaire. A doctor and a nurse performed a structured interview and graded the patient's symptoms with the same questions. The procedure was performed at both the start and the end of the treatment. A high concordance regarding symptom detection was seen between the patient, nurse and the doctor. The inter-rater test shows intraclass correlation coefficient (ICC) values above 0.60 in all scales. The internal reliability exceeded the lower limit (Cronbach a >0.70) for all scales. The test-retest gave acceptable reliability for all scales (ICC > or = 0.60). All scales indicated increased problems during radiotherapy. The questionnaire was proven to be valid for the evaluations of urinary and intestinal problems and for sexual function in PC patients. PMID- 11401104 TI - Follow-up of patients with haematological malignancies and their families in regional, rural and remote Queensland: the GPs' perspective. AB - In rural areas of Australia the range of health services is limited and major inequities exist in health care provision. Consequently, general practitioners (GPs) are forced to play a pivotal role as key providers of health and medical services. Patients with a haematological malignancy returning home from the lengthy ordeal of high-tech specialist treatment in the metropolitan area will have a strong need for competent holistic support. For these patients and their families, establishing a positive relationship with the local GP will be an important factor in their post-treatment rehabilitation. The findings presented in this article are from recent Queensland research that looks at the support provided by GPs in regional, rural, and remote areas for patients with a haematological malignancy after they return home from specialist treatment in the metropolitan area. The findings indicate the need for support and education for GPs coping with such patients. Newsletters and telephone contact are recorded as the most popular means of support by the GPs surveyed. PMID- 11401105 TI - Gabapentin for opiod-related myoclonus in cancer patients. AB - Chronic opioid medication has been found to cause myoclonus in patients taking it for cancer pain. Gabapentin seemed a likely candidate for the treatment of this myoclonus and has indeed proved useful, as illustrated in this paper by two case histories. PMID- 11401106 TI - The desire to be fashionable. Should we consider it a risk factor for infection in adolescents with acute leukaemia? AB - Three cases of skin infection, two with Pseudomonas aeruginosa and one with Staphylococcus aureus in three adolescents with acute leukaemia are described. In all cases the infection was clearly related to the latest fashion in shoes. This report underline the fact that factors related to everyday life and less frequently considered than those related to the underlying illness can also put increase the risk of developing severe infections in immunocompromised patients. PMID- 11401107 TI - Liposomal nystatin (L-NYS) in therapy of pulmonary aspergillosis refractory to conventional amphotericin B in cancer patients. PMID- 11401108 TI - A remarkable medical story: benefits of angiotensin-converting enzyme inhibitors in cardiac patients. AB - The development of angiotensin-converting enzyme inhibitors (ACE inhibitors) has been one of the most remarkable stories in the treatment of cardiovascular diseases. Angiotensin converting enzyme inhibitors have several acute and sustained hemodynamic effects that are beneficial in the presence of left ventricular (LV) dysfunction. They increase cardiac output and stroke volume and reduce systemic vascular resistance as well as pulmonary capillary wedge pressure. The hemodynamic benefits are associated with improvement in the signs and symptoms of congestive heart failure (CHF) as well as decreased mortality, regardless of the severity of CHF. In patients with asymptomatic LV dysfunction, therapy with ACE inhibitors prevented the development of CHF and reduced hospitalization and cardiovascular death. They also increase survival when administered early after an acute myocardial infarction (MI). Most recently, ACE inhibition was associated with improved clinical outcomes in a broad spectrum of high-risk patients with preserved LV function. The mechanism of ACE inhibitors benefits is multifactorial and includes prevention of progressive LV remodeling, prevention of sudden death and arrhythmogenicity and structural stability of the atherosclerotic process. Evidence suggests that ACE inhibitors are underutilized in patients with cardiovascular diseases. Efforts should be directed to prescribe ACE inhibitors to appropriate patients in target doses. It is reasonable to believe that ACE inhibitors have a class effect in the management of LV dysfunction with or without CHF and acute MI. Whether the same is true for ACE inhibitors in the prevention of ischemic events is not known yet. PMID- 11401109 TI - Chronic heart failure and micronutrients. AB - Heart failure (HF) is associated with weight loss, and cachexia is a well recognized complication. Patients have an increased risk of osteoporosis and lose muscle bulk early in the course of the disease. Basal metabolic rate is increased in HF, but general malnutrition may play a part in the development of cachexia, particularly in an elderly population. There is evidence for a possible role for micronutrient deficiency in HF. Selective deficiency of selenium, calcium and thiamine can directly lead to the HF syndrome. Other nutrients, particularly vitamins C and E and beta-carotene, are antioxidants and may have a protective effect on the vasculature. Vitamins B6, B12 and folate all tend to reduce levels of homocysteine, which is associated with increased oxidative stress. Carnitine, co-enzyme Q10 and creatine supplementation have resulted in improved exercise capacity in patients with HF in some studies. In this article, we review the relation between micronutrients and HF. Chronic HF is characterized by high mortality and morbidity, and research effort has centered on pharmacological management, with the successful introduction of angiotensin-converting enzyme inhibitors and beta-adrenergic antagonists into routine practice. There is sufficient evidence to support a large-scale trial of dietary micronutrient supplementation in HF. PMID- 11401110 TI - The effect of correction of mild anemia in severe, resistant congestive heart failure using subcutaneous erythropoietin and intravenous iron: a randomized controlled study. AB - OBJECTIVES: This is a randomized controlled study of anemic patients with severe congestive heart failure (CHF) to assess the effect of correction of the anemia on cardiac and renal function and hospitalization. BACKGROUND: Although mild anemia occurs frequently in patients with CHF, there is very little information about the effect of correcting it with erythropoietin (EPO) and intravenous iron. METHODS: Thirty-two patients with moderate to severe CHF (New York Heart Association [NYHA] class III to IV) who had a left ventricular ejection fraction (LVEF) of < or =40% despite maximally tolerated doses of CHF medications and whose hemoglobin (Hb) levels were persistently between 10.0 and 11.5 g% were randomized into two groups. Group A (16 patients) received subcutaneous EPO and IV iron to increase the level of Hb to at least 12.5 g%. In Group B (16 patients) the anemia was not treated. The doses of all the CHF medications were maintained at the maximally tolerated levels except for oral and intravenous (IV) furosemide, whose doses were increased or decreased according to the clinical need. RESULTS: Over a mean of 8.2+/-2.6 months, four patients in Group B and none in Group A died of CHF-related illnesses. The mean NYHA class improved by 42.1% in A and worsened by 11.4% in B. The LVEF increased by 5.5% in A and decreased by 5.4% in B. The serum creatinine did not change in A and increased by 28.6% in B. The need for oral and IV furosemide decreased by 51.3% and 91.3% respectively in A and increased by 28.5% and 28.0% respectively in B. The number of days spent in hospital compared with the same period of time before entering the study decreased by 79.0% in A and increased by 57.6% in B. CONCLUSIONS: When anemia in CHF is treated with EPO and IV iron, a marked improvement in cardiac and patient function is seen, associated with less hospitalization and renal impairment and less need for diuretics. PMID- 11401111 TI - Plasma N-terminal pro-brain natriuretic peptide and adrenomedullin: prognostic utility and prediction of benefit from carvedilol in chronic ischemic left ventricular dysfunction. Australia-New Zealand Heart Failure Group. AB - OBJECTIVES: We sought to assess plasma concentrations of the amino (N)-terminal portion of pro-brain natriuretic peptide (N-BNP) and adrenomedullin for prediction of adverse outcomes and responses to treatment in 297 patients with ischemic left ventricular (LV) dysfunction who were randomly assigned to receive carvedilol or placebo. BACKGROUND: Although neurohormonal status has known prognostic significance in heart failure, the predictive power of either N-BNP or adrenomedullin in chronic ischemic LV dysfunction has not been previously reported. METHODS: Plasma N-BNP and adrenomedullin were measured in 297 patients with chronic ischemic (LV) dysfunction before randomization to carvedilol or placebo, added to established treatment with a converting enzyme inhibitor and loop diuretic (with or without digoxin). The patients' clinical outcomes, induding mortality and heart failure events, were recorded for 18 months. RESULTS: Above-median N-BNP and adrenomedullin levels conferred increased risks (all p < 0.001) of mortality (risk ratios [95% confidence intervals]: 4.67 [2 10.9] and 3.92 [1.76-8.7], respectively) and hospital admission with heart failure (4.7 [2.2-10.3] and 2.4 [1.3-4.5], respectively). Both of these predicted death or heart failure independent of age, New York Heart Association functional class, LV ejection fraction, previous myocardial infarction or previous admission with heart failure. Carvedilol reduced the risk of death or heart failure in patients with above-median levels of N-BNP or adrenomedullin, or both, to rates not significantly different from those observed in patients with levels below the median value. CONCLUSIONS: In patients with established ischemic LV dysfunction, plasma N-BNP and adrenomedullin are independent predictors of mortality and heart failure. Carvedilol reduced mortality and heart failure in patients with higher pre-treatment plasma N-BNP and adrenomedullin. PMID- 11401112 TI - Effects of posture on cardiac autonomic nervous activity in patients with congestive heart failure. AB - OBJECTIVES: We aimed to clarify which recumbent position is preferred by patients with congestive heart failure (CHF) and to evaluate whether cardiac autonomic nervous activity is different among three recumbent positions (supine, left lateral decubitus, right lateral decubitus) in patients with CHF. BACKGROUND: It remains unclear whether cardiac autonomic nervous activity is different among three recumbent positions in patients with CHF. METHODS: We studied 17 male CHF patients (66+/-7 years) and 17 age- and gender-matched healthy subjects (66+/-7 years). Each subject underwent 24-h ambulatory electrocardiographic monitoring. A channel was used to record the CM5 lead, and another to record the signal of the patient's posture with use of a newly developed small-sized detector (3.2 cm x 3.2 cm). By using spectral analysis of heart rate variability, frequency-domain measures were calculated and compared among the three recumbent positions. Normalized high-frequency (HF: 0.15 to 0.40 Hz) power was used as an index of vagal activity and the low frequency (0.04 to 0.15 Hz)/HF power ratio was used as an index of sympathovagal balance. RESULTS: In patients with CHF, the time for the right lateral decubitus position was two-fold longer than that for the supine and left lateral decubitus positions. The increased cardiac sympathetic activity and decreased vagal tone in CHF patients were normalized in the right lateral decubitus position. CONCLUSIONS: The right lateral decubitus position in patients with CHF may be a self-protecting mechanism of attenuating the imbalance of cardiac autonomic nervous activity. PMID- 11401113 TI - Exercise performance in patients with end-stage heart failure after implantation of a left ventricular assist device and after heart transplantation: an outlook for permanent assisting? AB - OBJECTIVES: We sought to study exercise capacity at different points in time after left ventricular assist device (LVAD) implantation and subsequent heart transplantation (HTx). BACKGROUND: The lack of donor organs warrants alternatives for transplantation. METHODS: Repeat treadmill testing with respiratory gas analysis was performed in 15 men with a LVAD. Four groups of data are presented. In group A (n = 10), the exercise capacities at 8 weeks and 12 weeks after LVAD implantation were compared. In group B (n = 15), the data at 12 weeks are presented in more detail. In group C (n = 9), sequential analysis of exercise capacity was performed at 12 weeks after LVAD implantation and at 12 weeks and one year after HTx. In group D, exercise performance one year after HTx in patients with (n = 10) and without (n = 20) a previous assist device was compared. RESULTS: In group A, peak oxygen consumption (Vo2) increased from 21.3+/-3.8 to 24.2+/-4.8 ml/kg body weight per min (p < 0.003), accompanied by a decrease in peak minute ventilation/ carbon dioxide production (VE/Vco2) (39.4+/ 10.1 to 36.3+/-8.2; p < 0.03). In group B, peak Vo2 12 weeks after LVAD implantation was 23.0+/-4.4 ml/kg per min. In group C, levels of peak Vo2 12 weeks after LVAD implantation and 12 weeks and one year after HTx were comparable (22.8+/-5.3, 24.6+/-3.3 and 26.2+/-3.8 ml/kg per min, respectively; p = NS). In group D, there appeared to be no difference in percent predicted peak Vo2 in patients with or without a previous LVAD (68+/-13% vs. 74+/-15%; p < 0.37), although, because of the small numbers, the power of this comparison is limited (0.45 to detect a difference of 10%). CONCLUSIONS: Exercise capacity in patients with a LVAD increases over time; 12 weeks after LVAD implantation, Vo2 is comparable to that at 12 weeks and one year after HTx. Previous LVAD implantation does not seem to adversely affect exercise capacity after HTx. PMID- 11401114 TI - Circadian variation in the effects of aldosterone blockade on heart rate variability and QT dispersion in congestive heart failure. AB - OBJECTIVES: The study was designed to comprehensively evaluate the circadian effects of aldosterone blockade on autonomic tone and QT dispersion in chronic heart failure (CHF). BACKGROUND: Spironolactone therapy given in addition to angiotensin-converting enzyme inhibitors improved survival in CHF, but the mechanism of its benefit is uncertain. Experimental evidence suggests that aldosterone may have detrimental effects on the autonomic nervous system, especially during the morning hours. METHODS: Twenty-eight patients with New York Heart Association class II to IV CHF received spironolactone 50 mg daily and placebo for four weeks each in a double-blind crossover fashion. After each treatment phase, a full circadian assessment was undertaken of spironolactone's autonomic effects. The assessment included monitoring heart rate, QT dispersion, continuous Holter recordings, heart rate variability (HRV) and norepinephrine kinetics. RESULTS: Spironolactone significantly reduced all indices of QT dispersion. The reductions in QTcmax, QTd and QTcd were greatest at 6 AM. In addition, spironolactone had favorable autonomic effects, which were limited to the morning (6-10 AM), including heart rate reduction and an improvement in HRV. CONCLUSIONS: Spironolactone reduced heart rate and improved HRV and QT dispersion in CHF. Its effects were particularly prominent during the morning hours. PMID- 11401115 TI - Failure of aldosterone suppression despite angiotensin-converting enzyme (ACE) inhibitor administration in chronic heart failure is associated with ACE DD genotype. AB - OBJECTIVES: The objective of this study was to assess whether the angiotensin converting enzyme (ACE) gene insertion/deletion (I/D) polymorphism influences the adequacy of the neurohormonal response to ACE inhibitors in patients with chronic heart failure (CHF). BACKGROUND: The renin-angiotensin-aldosterone system (RAAS) plays an important role in the pathophysiology of CHF, and aldosterone levels closely relate to outcome in patients with CHF. Angiotensin-converting enzyme inhibitors suppress the RAAS, but a significant proportion of patients exhibit elevated serum levels of aldosterone despite long-term administration of apparently adequate doses of these agents. METHODS: We prospectively studied 132 patients with CHF (ejection fraction <45%) receiving long-term therapy with ACE inhibitors for over six months. Patients taking aldosterone antagonists were excluded from the study. "Aldosterone escape" was defined as being present when plasma aldosterone levels were above the normal range in our laboratory (>42 nmol/L). Patients were then divided into two subgroups according to the presence (group 1) or absence (group 2) of aldosterone escape. Genotype analysis for the ACE I/D polymorphism was performed by polymerase chain reaction. RESULTS: The prevalence of aldosterone escape in our patients was 10% (13/132). The two groups of patients did not differ regarding the dose of ACE inhibitor, diuretics and their renal function. There was a statistically significant different distribution of genotypes between the two groups, with a higher proportion of DD genotype in group 1 compared with group 2 (62% vs. 24%, p = 0.005). CONCLUSIONS: Patients with CHF with aldosterone escape have a higher prevalence of DD genotype compared with patients with aldosterone within the normal limits. Angiotensin converting enzyme gene polymorphism contributes to the modulation and adequacy of the neurohormonal response to long-term ACE-inhibitor administration in CHF. PMID- 11401116 TI - Accurate noninvasive estimation of pulmonary vascular resistance by Doppler echocardiography in patients with chronic failure heart failure. AB - OBJECTIVES: This study was undertaken to explore further the relationship between Doppler-derived parameters of pulmonary flow and pulmonary vascular resistance (PVR) and to determine whether PVR could be accurately estimated noninvasively from Doppler flow velocity measurements in patients with chronic heart failure. BACKGROUND: The assessment of PVR is of great importance in the management of patients with heart failure. However, because of the inconclusive and conflicting data available, Doppler estimation of PVR is still considered unreliable. METHODS: Simultaneous Doppler echocardiographic examination and right heart catheterization were performed in 63 consecutive sinus rhythm heart failure patients with severe left ventricular systolic dysfunction. Hemodynamic PVR was calculated with the standard formula. The following Doppler variables on pulmonary flow and tricuspid regurgitation velocity curve were correlated with PVR: maximal systolic flow velocity, pre-ejection period (PEP), acceleration time (AcT), ejection time, total systolic time (TT), velocity time integral, and right atrium-ventricular gradient. RESULTS: At univariate analysis, all variables except maximal systolic flow velocity and velocity time integral showed a significant, although weak, correlation with PVR. The best correlation found was between AcT and PVR (r = -0.68). By regression analysis, only PEP, AcT and TT entered into the final equation, with a cumulative r = 0.87. When the function (PEP/AcT)/TT was correlated with PVR, the correlation coefficient further improved to 0.96. Of note, this function prospectively predicted PVR (r = 0.94) after effective unloading manipulations. CONCLUSIONS: The analysis of Doppler derived pulmonary systolic flow is a reliable and accurate tool for estimating and monitoring PVR in patients with chronic heart failure due to left ventricular systolic dysfunction. PMID- 11401117 TI - Intravenous atrial natriuretic peptide prevents left ventricular remodeling in patients with first anterior acute myocardial infarction. AB - OBJECTIVES: The study evaluates the effect of atrial natriuretic peptide (ANP) compared with nitroglycerin (GTN) on left ventricular (LV) remodeling after first anterior acute myocardial infarction (AMI). BACKGROUND: Compared with GTN, ANP suppresses the renin-angiotensin-aldosterone system and endothelin-1 (ET-1), which stimulate LV remodeling. METHODS: Sixty patients with a first anterior AMI were randomly divided into the ANP (n = 30) or GTN (n = 30) groups after direct percutaneous transluminal coronary angioplasty. We evaluated LV ejection fraction (LVEF), end-diastolic volume index (LVEDVI) and end-systolic volume index (LVESVI) at the acute phase and after one month. We also measured neurohumoral factors during study drug infusion. RESULTS: There was no difference in the baseline characteristics or LVEF (46.9+/-1.0 vs. 46.8+/-1.3%) between the two groups. Although there was no difference in hemodynamics during the infusion periods, the LVEF was significantly improved after one month compared with the baseline value in both groups, but it was improved more in the ANP group than in the GTN group (54.6+/-1.1%, 50.8+/-1.3%, p < 0.05). Left ventricular enlargement was prevented in the ANP group (LVEDVI, 85.8+/-3.1 ml/m2 to 87.3+/-2.7 ml/m2; p = ns, LVESVI, 45.6+/-1.8 ml/m2 to 41.0+/-2.1 ml/m2, p < 0.05) but not in the GTN group (LVEDVI, 86.2+/-4.1 to 100.2+/-3.7, p < 0.01; LVESVI, 46.3+/-2.8 ml/m2 to 51.1+/-3.0 ml/m2, p = ns). During the infusion, ANP suppressed plasma levels of aldosterone, angiotensin II and ET-1 compared with GTN. CONCLUSIONS: These findings indicate that in patients with a first anterior AMI, an ANP infusion can prevent LV remodeling better than can GTN, and effectively suppresses aldosterone, angiotensin II and ET-1. PMID- 11401118 TI - Primary angioplasty versus intravenous thrombolysis in acute myocardial infarction: can we define subgroups of patients benefiting most from primary angioplasty? Results from the pooled data of the Maximal Individual Therapy in Acute Myocardial Infarction Registry and the Myocardial Infarction Registry. AB - OBJECTIVES: We sought to determine the effectiveness of primary angioplasty compared with thrombolysis in clinical practice. BACKGROUND: In clinical practice, primary angioplasty for the treatment of acute myocardial infarction (AMI) has not yet been proven more effective than intravenous thrombolysis, nor have subgroups of patients been identified who would perhaps benefit from primary angioplasty. METHODS: The pooled data of two AMI registries--the Maximal Individual TheRapy in Acute myocardial infarction (MITRA) study and the Myocardial Infarction Registry (MIR)--were analyzed. A total of 9,906 lytic eligible patients with AMI, with a pre-hospital delay of < or =12 h, were treated with either primary angioplasty (n = 1,327) or thrombolysis (n = 8,579). RESULTS: Despite differences in the patients' characteristics and concomitant diseases between the two groups, the prevalence of adverse risk factors was balanced. Univariate analysis of hospital mortality showed a more favorable course for patients treated with primary angioplasty: 6.4% versus 11.3% (odds ratio [OR] 0.54, 95% confidence interval [CI] 0.43 to 0.67). This was confirmed by logistic regression analysis (multivariate OR 0.58, 95% CI 0.44 to 0.77). Primary angioplasty was associated with a lower mortality in all subgroups analyzed. We observed a significant correlation between mortality and absolute risk reduction (r = 0.82, p < 0.0001) in the different subgroups: as mortality increased, there was an increase in absolute benefit of primary angioplasty compared with thrombolysis. CONCLUSIONS: These large registry data showed the effect of primary angioplasty to be more favorable than thrombolysis for the treatment of patients with AMI in clinical practice. This effect was not restricted to special subgroups of patients. As mortality increased, the absolute benefit of primary angioplasty also increased. PMID- 11401119 TI - The effectiveness of primary PTCA: does patient risk matter? PMID- 11401120 TI - Myocardial infarction after vascular surgery: the role of prolonged stress induced, ST depression-type ischemia. AB - OBJECTIVES: The goal of this study was to investigate the nature of the association between silent ischemia and postoperative myocardial infarction (PMI). BACKGROUND: Silent ischemia predicts cardiac morbidity and mortality in both ambulatory and postoperative patients. Whether silent stress-induced ischemia is merely a marker of extensive coronary artery disease or has a closer association with infarction has not been determined. METHODS: In 185 consecutive patients undergoing vascular surgery, we correlated ischemia duration, as detected on a continuous 12-lead ST-trend monitoring during the period 48 h to 72 h after surgery, with cardiac troponin-I (cTn-I) measured in the first three postoperative days and with postoperative cardiac outcome. Postoperative myocardial infarction was defined as cTn-I >3.1 ng/ml accompanied by either typical symptoms or new ischemic electrocardiogram (ECG) findings. RESULTS: During 11,132 patient-hours of monitoring, 38 patients (20.5%) had 66 transient ischemic events, all but one denoted by ST-segment depression. Twelve patients (6.5%) sustained PMI; one of those patients died. All infarctions were non-Q-wave and were detected by a rise in cTn-I during or immediately after prolonged, ST depression-type ischemia. The average duration ofischemia in patients with PMI was 226+/-164 min (range: 29 to 625), compared with 38+/-26 min (p = 0.0000) in 26 patients with ischemia but not infarction. Peak cTn-I strongly correlated with the longest, as well as cumulative, ischemia duration (r = 0.83 and r = 0.78, respectively). Ischemic ECG changes were completely reversible in all but one patient who had persistent new T wave inversion. All ischemic events culminating in PMI were preceded by an increase in heart rate (delta heart rate = 32+/-15 beats/min), and most (67%) of them began at the end of surgery and emergence from anesthesia. CONCLUSIONS: Prolonged, ST depression-type ischemia progresses to MI and is strongly associated with the majority of cardiac complications after vascular surgery. PMID- 11401121 TI - Pre-infarction angina elicits greater myocardial viability on reperfusion after myocardial infarction: a dobutamine stress echocardiographic study. AB - OBJECTIVES: We sought to evaluate myocardial viability (inotropic reserve) after myocardial infarction (MI) and its relationship with the presence of unstable pre infarction angina (PIA). BACKGROUND: Several studies have suggested that PIA can limit infarct size, but it is not known whether PIA can elicit myocardial viability after an acute MI, with left ventricular function improvement. METHODS: Before discharge from the hospital, 91 patients with a reperfused MI (either fibrinolysis or primary coronary angioplasty) had low-dose dobutamine echocardiography performed to assess the myocardial inotropic reserve of the infarct-related area. RESULTS: Twenty-nine patients (31.9%) had PIA in the 24-h period before the onset of MI. Nine patients were treated with primary coronary angioplasty: five (8.1%) in the group with PIA and four (13.8%) in the group without PIA. There were no other significant differences in the baseline characteristics of the patients. There were more viable segments in patients with PIA (44.9% vs. 30.7%, p = 0.007), and the number of patients with significant viability was higher in the PIA group (73.9% vs. 46.3%, p = 0.026). This occurred despite a similar number of segments with segmental wall abnormalities at baseline in both groups (46.1% vs. 46.9%, p = NS). CONCLUSIONS: The presence of previous unstable PIA induces greater myocardial viability of the infarct-related area upon reperfusion and, as such, could have considerable therapeutic and clinical implications. PMID- 11401122 TI - Stable angina and acute coronary syndromes are associated with nitric oxide resistance in platelets. AB - OBJECTIVES: The study examined possible clinical determinants of platelet resistance to nitric oxide (NO) donors in patients with stable angina pectoris (SAP) and acute coronary syndromes (ACS), relative to nonischemic patients and normal subjects. BACKGROUND: We have shown previously that platelets from patients with SAP are resistant to the antiaggregating effects of nitroglycerin (NTG) and sodium nitroprusside (SNP). METHODS: Extent of adenosine diphosphate (1 micromol/liter)-induced platelet aggregation (impedance aggregometry in whole blood) and inhibition of aggregation by NTG (100 micromol/liter) and SNP (10 micromol/liter) were compared in normal subjects (n = 43), nonischemic patients (those with chest pain but no fixed coronary disease, (n = 35) and patients with SAP (n = 82) or ACS (n = 153). Association of NO resistance with coronary risk factors, coronary artery disease (CAD), intensity of angina and current medication was examined by univariate and multivariate analyses. RESULTS: In patients with SAP and ACS as distinct from nonischemic patients and normal subjects, platelet aggregability was increased (both p < 0.01), and inhibition of aggregation by NTG and SNP was decreased (both p < 0.01). Multivariate analysis revealed that NO resistance occurred significantly more frequently with ACS than with SAP (odds ratio [OR] 2.3:1), and was less common among patients treated with perhexiline (OR 0.3:1) or statins (OR 0.45:1). Therapy with other antianginal drugs, extent of CAD, intensity of angina and coronary risk factors were not associated with variability in platelet responsiveness to NO donor. CONCLUSIONS: Patients with symptomatic ischemic heart disease, especially ACS, exhibit increased platelet aggregability and decreased platelet responsiveness to the antiaggregatory effects of NO donors. The extent of NO resistance in platelets is not correlated with coronary risk factors. Pharmacotherapy with perhexiline and/or statins may improve platelet responsiveness to NO. PMID- 11401123 TI - A randomized double-blind placebo-controlled trial of the effect of homocysteine lowering therapy with folic acid on endothelial function in patients with coronary artery disease. AB - OBJECTIVES: This study was designed to determine the effects of folic acid therapy on endothelial function in patients with coronary artery disease (CAD). BACKGROUND: Hyperhomocysteinemia, a risk factor for CAD, may cause atherosderosis by oxidative endothelial injury. Folic acid reduces plasma homocysteine, but the effect on adverse vascular events is unknown. METHODS: In a double-blind placebo controlled trial, 90 patients (mean age [range] 63 [46 to 79] years, 79 men) with CAD were randomized to either folic acid 5 mg or placebo daily for 12 weeks. Endothelial function was assessed by measuring: 1) flow-mediated endothelium dependent dilation (EDD) of the brachial artery; 2) combined serum nitrite/nitrate (NOx) concentrations and; 3) plasma von Willebrand factor (vWF) concentration. RESULTS: At the end of the study, plasma homocysteine was lower in the folic acid group compared with the placebo group (mean [95% confidence interval] 9.3 (8.5 to 10.1) vs. 12.3 [11.3 to 13.4] micromol/l, p < 0.001). Although there were no significant differences in EDD, serum NOx or plasma vWF between the two groups, there was a greater increase in EDD from baseline in the folic acid group compared to placebo (1.2 [0.7 to 1.8] vs. 0.4 [-0.3 to 1.1]%, p = 0.07). CONCLUSIONS: Folic acid reduced plasma homocysteine and was associated with a trend toward improved endothelial function in patients with CAD. The absence of an unequivocally positive result may have been due to inadequate sample size or chance. This reinforces the need for the results of large randomized controlled trials before the implementation of routine folic acid supplementation. PMID- 11401124 TI - Plaque distribution and vascular remodeling of ruptured and nonruptured coronary plaques in the same vessel: an intravascular ultrasound study in vivo. AB - OBJECTIVES: This study was designed to identify potential differences between the intravascular ultrasound (IVUS) characteristics of spontaneously ruptured and nonruptured coronary plaques. BACKGROUND: The identification of vulnerable plaques in vivo may allow targeted prevention of acute coronary events and more effective evaluation of novel therapeutic approaches. METHODS: Intravascular ultrasound was used to identify 29 ruptured plaques in arteries containing another nonruptured plaque in an adjacent segment. Intravascular ultrasound characteristics of these plaques were compared with plaques of computer-matched controls without evidence of plaque rupture. Plaque distribution was assessed by measuring the eccentricity of lumen location (inside the total vessel). Lumen cross-sectional area narrowing was calculated as [1 - (target/reference lumen area)] x 100%. A remodeling index was calculated as lesion/reference arterial area (>1.05 = compensatory enlargement, <0.95 = shrinkage). RESULTS: Among the three groups of plaques, there was no significant difference in quantitative angiographic parameters, IVUS reference dimensions and IVUS lumen cross-sectional area narrowing. There was a difference in plaque distribution; lumen location by IVUS was significantly more eccentric in ruptured than in nonruptured (p = 0.002) and control plaques (p < 0.0001). The arc of disease-free vessel wall was larger in ruptured than in control plaques (p < 0.0001). The remodeling pattern of ruptured and nonruptured plaques differed significantly from that of the control plaques (p = 0.0001 and 0.003); compensatory enlargement was found in 66%, 48%, and 17%, whereas shrinkage was found in 7%, 10% and 48%, respectively. CONCLUSIONS: Intravascular ultrasound assessment of plaque distribution and vascular remodeling may help to classify plaques with the highest probability of spontaneous rupture. PMID- 11401125 TI - Antibody against oxidized low density lipoprotein may predict progression or regression of atherosclerotic coronary artery disease. AB - OBJECTIVES: This study aimed to elucidate whether an antibody against oxidized low density lipoprotein (anti-Ox-LDL) could predict short-term coronary artery atherosclerotic lesion progression. BACKGROUND: It is still controversial whether higher levels of the anti-Ox-LDL titer are associated with atherosclerotic coronary artery disease. METHODS: In 52 patients undergoing coronary angioplasty and six-month follow-up angiography, we performed quantitative coronary angiographic analysis of a lesion on a branch away from the intervention site vessel and assessed lesion progression or regression using the Progression Regression score calculated as the baseline minimal lumen diameter minus the follow-up minimal lumen diameter. The serum anti-Ox-LDL titer was measured using an enzyme-linked immunosorbent assay method just before the initial angiography in all patients. RESULTS: The anti-Ox-LDL titer was 16.6+/-1.5 AcU/ml in the progression group (Progression-Regression score >0.15 mm; n = 20), which was significantly higher (p < 0.001) than the value of 9.5+/-1.2 in the regression group (< or =-0.15 mm; n = 14) and also higher (p < 0.01) than the value of 11.4+/-1.3 in the no-change group (-0.15 to 0.15 mm; n = 18). The Progression Regression score was correlated with the antibody titer in all patients (r = 0.56, p < 0.001). Multiple regression analysis showed that the Progression Regression score was independently correlated with the antibody titer (r = 0.44, p < 0.01) as well as lipoprotein (a) (r = 0.33, p < 0.05). CONCLUSIONS: Anti-Ox LDL may be an independent predictor of coronary atherosclerotic lesion progression in the short term. PMID- 11401126 TI - Percutaneous and surgical interventions for in-stent restenosis: long-term outcomes and effect of diabetes mellitus. AB - OBJECTIVE: We examined long-term outcomes of patients with in-stent restenosis (ISR) who underwent different percutaneous interventions at the discretion of individual operators: balloon angioplasty (BA), repeat stent or rotational atherectomy (RA). We also examined long-term outcomes of patients with ISR who underwent coronary artery bypass surgery (CABG). BACKGROUND: In-stent restenosis remains a challenging problem, and its optimal management is still unknown. METHODS: Symptomatic patients (n = 510) with ISR were identified using cardiac catheterization laboratory data. Management for ISR included BA (169 patients), repeat stenting (117 patients), RA (107 patients) or CABG (117 patients). Clinical outcome events of interest included death, myocardial infarction, target vessel revascularization (TVR) and a combined end point of these major adverse cardiovascular events (MACE). Mean follow-up was 19+/-12 months (range = 6 to 61 months). RESULTS: Patients with ISR treated with repeat stent had significantly larger average post-procedure minimal lumen diameter compared with BA or RA (3.3+/-0.4 mm vs. 3.0+/-0.4 vs. 2.9+/-0.5, respectively, p < 0.05). Incidence of TVR and MACE were similar in the BA, stent and RA groups (39%, 40%, 33% for TVR and 43%, 40%, 33% for MACE, p = NS). Patients with diabetes who underwent RA had similar outcomes as patients without diabetes, while patients with diabetes who underwent BA or stent had worse outcomes than patients without diabetes. Patients who underwent CABG for ISR, mainly because of the presence of multivessel disease, had significantly better outcomes than any percutaneous treatment (8% for TVR and 23% for MACE). CONCLUSIONS: In this large cohort of patients with ISR and in the subset of patients without diabetes, long-term outcomes were similar in the BA, repeat stent and RA groups. Tissue debulking with RA yielded better results only in diabetic patients. Bypass surgery for patients with multivessel disease and ISR provided the best outcomes. PMID- 11401127 TI - Achieving optimal results with standard balloon angioplasty: can baseline and angiographic variables predict stent-like outcomes? AB - OBJECTIVES: To predict which patients might not require stent implantation, we identified clinical and angiographic characteristics associated with repeat revascularization after standard balloon angioplasty. BACKGROUND: Stents reduce the risk of repeat revascularization but are costly and may lead to in-stent restenosis, which remains difficult to treat. Identification of patients at low risk for repeat revascularization may allow clinicians to reserve stents for patients most likely to benefit. METHODS: Data from five interventional trials (5,146 patients) were pooled for analysis. We identified patients with optimal angiographic results (final diameter stenosis < or =30% and no dissection) after balloon angioplasty and determined the multivariable predictors of repeat revascularization. RESULTS: Optimal angiographic results were achieved in 18% of patients after angioplasty. The repeat revascularization rate at six months was lower for patients with optimal results (20% vs. 26%, p < 0.001) but still higher than observed in stent trials. Independent predictors of repeat revascularization were female gender (odds ratio [OR] 1.67, p = 0.01), lesion length > or =10 mm (OR 1.62, p = 0.03) and proximal left anterior descending coronary artery lesions (OR 1.62, p = 0.03). For the 8% of patients with optimal angiographic results and none of these risk factors, the repeat revascularization and target vessel revascularization rates were 14% and 8% respectively, similar to rates after stent implantation. Cost analysis estimated that $78 million per year might be saved in the U.S. with a provisional stenting strategy using these criteria compared with elective stenting. CONCLUSIONS: A combination of baseline characteristics and angiographic results can be used to identify a small group of patients at very low risk for repeat revascularization after balloon angioplasty. Provisional stenting for these low risk patients could substantially reduce costs without compromising clinical outcomes. PMID- 11401128 TI - Exercise training intervention after coronary angioplasty: the ETICA trial. AB - OBJECTIVES: The goal of this study was to determine the effects of exercise training (ET) on functional capacity and quality of life (QOL) in patients who received percutaneous transluminal coronary angioplasty (PTCA) or coronary stenting (CS), the effects on the restenosis rate and the outcome. BACKGROUND: It is unknown whether ET induces beneficial effects after coronary angioplasty. METHODS: We studied 118 consecutive patients with coronary artery disease (mean age 57+/-10 years) who underwent PTCA or CS on one (69%) or two (31%) native epicardial coronary arteries. Patients were randomized into two matched groups. Group T (n = 59) was exercised three times a week for six months at 60% of peak VO2. Group C (n = 59) was the control group. RESULTS: Only trained patients had significant improvements in peak VO2 (26%, p < 0.001) and quality of life (26.8%, p = 0.001 vs. C). The angiographic restenosis rate was unaffected by ET (T: 29%; C: 33%, P = NS) and was not significantly different after PTCA or CS. However, residual diameter stenosis was lower in trained patients (-29.7%, p = 0.045). In patients with angiographic restenosis, thallium uptake improved only in group T (19%; p < 0.001). During the follow-up (33+/-7 months) trained patients had a significantly lower event rate than controls (11.9 vs. 32.2%, RR: 0.71, 95% confidence interval [CI]: 0.60 to 0.91, p = 0.008) and a lower rate of hospital readmission (18.6 vs. 46%, RR: 0.69, 95% CI: 0.55 to 0.93, p < 0.001). CONCLUSIONS: Moderate ET improves functional capacity and QOL after PTCA or CS. During the follow-up, trained patients had fewer events and a lower hospital readmission rate than controls, despite an unchanged restenosis rate. PMID- 11401129 TI - Value of programmed ventricular stimulation for prophylactic internal cardioverter-defibrillator implantation in postinfarction patients preselected by noninvasive risk stratifiers. AB - OBJECTIVES: The aim of this prospective study was to evaluate the role of programmed ventricular stimulation (PVS) after noninvasive risk stratification to identify a subgroup of acute myocardial infarction (AMI) survivors considered at risk for ventricular arrhythmias and whether these patients could benefit from internal cardioverter-defibrillators (ICDs). BACKGROUND: The predictive value of noninvasive and invasive risk stratifiers after AMI has been questioned. The question of whether the group of patients with inducible monomorphic ventricular tachycardia (VT) after AMI could profit from ICD implantation is unanswered. METHODS: A consecutive series of 1,436 AMI survivors was screened noninvasively by Holter monitoring, heart rate variability, ventricular late potentials, and ejection fraction. A subgroup of 248 patients (17.3%) were identified as high risk patients and scheduled for PVS. Due to the study design, 54 patients >75 years were excluded; thus, 194 patients were eligible for PVS. Triple extrastimuli at two paced cycle lengths (600 ms and 400 ms) were applied. RESULTS: In a subgroup of 98 (51%) high-risk patients, PVS was performed; 21 patients had an abnormal response, and in 20 patients an ICD was implanted. During a mean follow-up of 607 days the arrhythmic event rate (sudden cardiac death, symptomatic VT, cardiac arrest) was 33% with a positive electrophysiological test versus 2.6% (p < 0.0001) with a negative electrophysiological test. A subgroup of 96 high-risk patients declined electrophysiological study. In this nonconsent group, cardiac mortality (combined sudden and nonsudden) was significantly higher (log-rank chi-square 9.38, p = 0.0022, relative risk 4.7, 1.6 to 13.9) compared to the group guided by electrophysiological testing and consecutive ICD implantation. CONCLUSIONS: After a two-step risk stratification, PVS is helpful in selecting a subgroup of AMI survivors without spontaneous ventricular arrhythmias who benefit from prophylactic ICD implantation. PMID- 11401130 TI - Is programmed stimulation in survivors of myocardial infarction helpful? PMID- 11401131 TI - Recurrence of symptomatic ventricular arrhythmias in patients with implantable cardioverter defibrillator after the first device therapy: implications for antiarrhythmic therapy and driving restrictions. CARE Group. AB - OBJECTIVES: The purpose of this study was to investigate whether clinical or electrophysiologic characteristics could predict initial and subsequent implantable cardioverter defibrillator (ICD) therapy. BACKGROUND: Identification of markers to predict subsequent ICD therapy and symptoms after the first event could affect patient management. METHODS: We analyzed baseline and follow-up data on 125 ICD patients followed for 408+/-321 days. Medications and ICD programming were not changed after first ICD therapy. RESULTS: Implantable cardioverter defibrillator therapy occurred in 58 patients (46%). Clinical features were as follows: mean left ventricular ejection fraction (LVEF) 29%+/-15%; coronary artery disease 84%; presenting arrhythmia with sustained monomorphic ventricular tachycardia (SMVT) in 68%. In a multivariate analysis the relative risk for ICD therapy in patients presenting with SMVT versus cardiac arrest (CA) was 2.57 (range, 1.32 to 5.01), and for patients with LVEF < or =25%, 1.95 (1.11 to 3.45), respectively (p < 0.05). Implantable cardioverter defibrillator therapy was not predicted by any other variable. Forty-six patients had second ICD therapy. Mean time to second ICD therapy was only 66+/-93 days compared with 138+/-168 days for first ICD therapy (p < 0.05). No predictor for second ICD therapy was found. Regarding symptoms, impaired consciousness during initial ICD therapy was predicted only by SMVT cycle length <250 ms at electrophysiologic testing. In contrast, symptoms were similar between first and second ICD therapy (p = 0.0001). Of note, ventricular tachycardia cycle length preceding first and second ICD therapy was similar (r = 0.76, p = 0.001). CONCLUSIONS: First ICD therapy tends to occur in patients presenting with SMVT and LVEF < or =25%. Subsequent therapy occurs sooner and is unpredictable, suggesting that antiarrhythmic drug therapy should be considered after the first symptomatic ICD therapy. Symptoms during first ICD therapy predict subsequent symptoms, and patients presenting with SMVT and asymptomatic first ICD therapy are at very low risk for future syncopal ICD therapy. PMID- 11401132 TI - Three-year follow-up of patients with right bundle branch block and ST segment elevation in the right precordial leads: Japanese Registry of Brugada Syndrome. Idiopathic Ventricular Fibrillation Investigators. AB - OBJECTIVES: We sought to determine the prevalence of right bundle branch block (RBBB) and ST segment elevation in the working Japanese population, as well as the event rate during a three-year prospective follow-up period. BACKGROUND: A poor prognosis of RBBB and ST segment elevation has been reported in Europe and South America, even in asymptomatic patients; however, a large population of asymptomatic patients with sporadic RBBB and ST segment elevation has not been studied. METHODS: Ten thousand 12-lead electrocardiograms (ECGs) were obtained during annual check-ups of working adults in the Tokyo area. This three-year prospective follow-up study consisted of 105 patients, including 20 with ventricular fibrillation, 18 with syncope and 67 who were asymptomatic. They were registered from 46 institutions in Japan. RESULTS: The prevalence of ECG abnormalities in working adults was 0.16%. A coved-type ST segment elevation was related to a history of cardiac events, and 18% of registered patients had PR prolongation and 9.5% had left-axis deviation. The cumulative cardiac event-free rate was 67.6% in the symptomatic group and 93.4% in the asymptomatic group (p = 0.0004) after three years. CONCLUSIONS: The recurrence rate of cardiac events in symptomatic patients was similar to that reported previously, but it was very low in sporadic asymptomatic patients. The ECG findings may help us to select patients for further examination and more accurate evaluation of their prognoses. PMID- 11401133 TI - Diagnostic value of history in patients with syncope with or without heart disease. AB - OBJECTIVES: We sought to establish what historical findings are predictive of the cause of syncope. BACKGROUND: The clinical features of the various types of syncope have not been systematically investigated. METHODS: Three hundred forty one patients with syncope were prospectively evaluated. Each patient was interviewed using a standard questionnaire. A cause of syncope was assigned using standardized diagnostic criteria. RESULTS: A cardiac cause of syncope was established in 23% of the patients, a neurally mediated cause in 58% and a neurologic or psychiatric cause in 1%, and in the remaining 18%, the cause of syncope remained unexplained. In a preliminary analysis including age, gender and the presence of suspected or certain heart disease after the initial evaluation, only heart disease was an independent predictor of a cardiac cause of syncope (odds ratio 16, p = 0.00001), with a sensitivity of 95% and a specificity of 45%. In contrast, the absence of heart disease allowed us to exclude a cardiac cause of syncope in 97% of the patients. In patients with certain or suspected heart disease, the most specific predictors of a cardiac cause were syncope in the supine position or during effort, blurred vision and convulsive syncope. Significant and specific predictors of a neurally mediated cause were time between the first and last syncopal episode >4 years, abdominal discomfort before the loss of consciousness and nausea and diaphoresis during the recovery phase. In the patients without heart disease, palpitation was the only significant predictor of a cardiac cause. CONCLUSIONS: The presence of suspected or certain heart disease after the initial evaluation is a strong predictor of a cardiac cause of syncope. A few historical findings are useful to predict cardiac and neurally mediated syncope in patients with and without heart disease. PMID- 11401134 TI - Post-prandial remnant lipids impair arterial compliance. AB - OBJECTIVES: We sought to examine the effects of plasma lipids, especially in remnants after a fat meal, on systemic arterial compliance (SAC), a newly recognized cardiovascular risk factor. BACKGROUND: Post-prandial remnants correlate with coronary heart disease events through mechanisms that may include vascular dysfunction, although the effect on SAC has not been studied. METHODS: Systemic arterial compliance was measured non-invasively over 6 h after a fat meal in 16 subjects with varying plasma triglyceride levels. Changes were related to rises in plasma lipids and remnant lipids. Systemic arterial compliance was measured in 20 subjects after a control low-fat meal. RESULTS: The fat meal induced increments in plasma triglyceride and remnant cholesterol and triglyceride (respectively +54%, 50% and 290% at 3 h, analysis of variance <0.001). Systemic arterial compliance fell at 3 h and 6 h by 25% and 27% (analysis of variance <0.001). Baseline SAC correlated significantly with all lipid concentrations at 0, 3 h and 6 h, but only with triglyceride on stepwise regression analysis. The SAC response to the low-fat meal was very small and not significant. CONCLUSIONS: This is the first demonstration of SAC becoming impaired after a fat meal. Remnant lipids and plasma total triglyceride appeared to contribute to the fall in SAC. PMID- 11401135 TI - Difference in the respiratory variation between pulmonary venous and mitral inflow Doppler velocities in patients with constrictive pericarditis with and without atrial fibrillation. AB - OBJECTIVES: The goal of this study was to evaluate the difference in the respiratory change from expiration to inspiration (%E) between pulsed Doppler mitral inflow (MV) and pulmonary venous flow (PV) velocities in patients with constrictive pericarditis (CP) and to describe the influence of atrial fibrillation (AF). BACKGROUND: The difference in %E between MV and PV velocities as well as the influence of AF on %E has not been well described. METHODS: Pulsed wave Doppler transesophageal echocardiography (TEE) was performed with respiratory monitoring in 31 patients with CP and sinus rhythm (SR) and in 10 patients with CP and AF. The MV early (E) and late diastolic (A) velocities and their velocity time integral (VTI) as well as PV systolic (S) and diastolic (D) velocities and their VTI were measured. RESULTS: Regardless of the cardiac rhythm: 1) The MV-E velocity and E-VTI as well as PV-D velocity and D-VTI significantly decreased from expiration to inspiration; 2) the %E in PV-D velocity (27% in SR and 35% in AF) and D-VTI (38% in SR and 45% in AF) was significantly greater than that in MV-E velocity (18% in SR and 15% in AF) and E VTI (21% in SR and 19% in AF), respectively; 3) the PV S/D and S/D-VTI significantly increased from expiration to inspiration. CONCLUSIONS: A significant respiratory variation was observed in both MV and PV velocities in CP, not only in patients with SR but also in those with AF. Moreover, the %E was greater in the PV velocities than it was in the MV velocities. Evaluation of the %E in the PV velocities using TEE can be a sensitive diagnostic strategy for evaluation of patients with CP, even in patients with AF. PMID- 11401136 TI - The impact of diabetes on left ventricular filling pattern in normotensive and hypertensive adults: the Strong Heart Study. AB - OBJECTIVES: We sought to determine the effect of diabetes mellitus (DM) on left ventricular (LV) filling pattern in normotensive (NT) and hypertensive (HTN) individuals. BACKGROUND: Diastolic abnormalities have been extensively described in HTN but are less well characterized in DM, which frequently coexists with HTN. METHODS: We analyzed the transmitral inflow velocity profile at the mitral annulus in four groups from the Strong Heart Study: NT-non-DM (n = 730), HTN-non DM (n = 394), NT-DM (n = 616) and HTN-DM (n = 671). The DM subjects were further divided into those with normal filling pattern (n = 107) and those with abnormal relaxation (AbnREL) (n = 447). RESULTS: The peak E velocity was lowest in HTN-DM, intermediate in NT-DM and HT-non-DM and highest in the NT-non-DM group (p < 0.001), with a reverse trend seen for peak A velocity (p < 0.001). In multivariate analysis, E/A ratio was lowest in HTN-DM and highest in NT-non-DM, with no difference between NT-DM and HTN-non DM (p < 0.001). Likewise, mean atrial filling fraction and deceleration time were highest in HTN-DM, followed by HTN-non-DM or NT-DM and lowest in NT-non-DM (both p < 0.05). Among DM subjects, those with AbnREL had higher fasting glucose (p = 0.03) and hemoglobin A1C (p = 0.04). CONCLUSIONS: Diabetes mellitus, especially with worse glycemic control, is independently associated with abnormal LV relaxation. The severity of abnormal LV relaxation is similar to the well-known impaired relaxation associated with HTN. The combination of DM and HTN has more severe abnormal LV relaxation than groups with either condition alone. In addition, AbnREL in DM is associated with worse glycemic control. PMID- 11401137 TI - Effectiveness of beta-blocker therapy after acute myocardial infarction in elderly patients with chronic obstructive pulmonary disease or asthma. AB - OBJECTIVES: We evaluated the use and effectiveness of beta-blocker therapy after acute myocardial infarction (AMI) for elderly patients with chronic obstructive pulmonary disease (COPD) or asthma. BACKGROUND: Because patients with COPD and asthma have largely been excluded from clinical trials of beta-blocker therapy for AMI, the extent to which these patients would benefit from beta-blocker therapy after AMI is not well defined. METHODS: Using data from the Cooperative Cardiovascular Project, we examined the relationship between discharge use of beta-blockers and one-year mortality in patients with COPD or asthma who were not using beta-agonists, patients with COPD or asthma who were concurrently using beta-agonists and patients with evidence of severe disease (use of prednisone or previous hospitalization for COPD or asthma) compared with patients without COPD or asthma. RESULTS: Of 54,962 patients without contraindications to beta blockers, patients with COPD or asthma (20%) were significantly less likely to be prescribed beta-blockers at discharge after AMI. After adjusting for demographic and clinical factors, we found that beta-blockers were associated with lower one year mortality in patients with COPD or asthma who were not on beta-agonist therapy (relative risk [RR] = 0.85, 95% confidence interval [CI] 0.73 to 1.00), similar to patients without COPD or asthma (RR = 0.86, 95% CI 0.81 to 0.92). A survival benefit for beta-blockers was not found among patients concurrently using beta-agonists or with severe COPD or asthma. CONCLUSIONS: Beta-blocker therapy after AMI may be beneficial for COPD or asthma patients with mild disease. A survival benefit was not found for elderly AMI patients with more severe pulmonary disease. PMID- 11401138 TI - The importance of age and obesity on the relation between diabetes and left ventricular mass. AB - OBJECTIVES: The study investigated the relation of age with diabetes, obesity and hypertension on left ventricular mass (LVM). BACKGROUND: Epidemiological studies demonstrate a general rise of LVM with aging, but whether this phenomenon is independent or a function of coexisting diseases that accompany the aging process is unclear. Although obesity, hypertension and diabetes often coexist and increase in prevalence with age, studies of LVM in diabetics have been reported in mostly nonobese populations, and with little regard to the age-hypertension obesity interactions and effects on LVM. METHODS: We prospectively measured LVM in 875 consecutive, mostly obese individuals (673 men, 202 women). Clinical data were obtained by chart review and clinical history. Echocardiographic measurements of LVM (American Society of Echocardiography criteria) were calculated using the Devereux formula and corrected for height2.7 (LVM/Ht). RESULTS: Mean age was 49.3+/-12.3 years, body mass index 33.3+/-8.0 kg/m2, and LVM/Ht2.7 41.7+/-13.4 g/m2.7. Of the total cohort, 673 patients were men, 519 obese, 228 hypertensive, and 52 diabetic. Of the 519 obese, 183 were hypertensive and 44 were diabetic (22 of those were hypertensive). Of the 228 hypertensives, 183 were obese and 26 were diabetic. On multivariate analysis, obesity (p = 0.0001), age (p = 0.0001), hypertension (p = 0.0003) and diabetes (p = 0.62) were all independently associated with LVM/Ht2.7. Obesity was the most potent independent predictor of LVM/Ht2.7, associated with an increase of 8.1 g/m2.7 in LVM/Ht2.7. In diabetics, obesity had a synergistic effect on LVM/Ht2.7 (p = 0.006), which was further amplified by age (p = 0.03). CONCLUSIONS: Age, obesity, hypertension and diabetes are all independent determinants of LVM. The magnitude of the effect of diabetes on LVM is mainly consequent to a significant interaction of diabetes with obesity and age. PMID- 11401139 TI - Quality of life in aortic valve replacement: pulmonary autografts versus mechanical prostheses. AB - OBJECTIVES: We sought to determine whether the quality of life (QoL) is different in patients after aortic valve replacement with mechanical prostheses or pulmonary autografts. BACKGROUND: Quality of life after mechanical valve replacement may be affected by the risk of thromboembolism and anticoagulation, and after autograft implantation, by the risk of degeneration and re-operation especially of the homograft. METHODS: Two groups of 40 patients each--one after the autograft procedure (group I) and one after mechanical valve implantation (group II)--were matched for age, gender and length of follow-up. At latest follow-up, all patients underwent routine echocardiography, the short-form health survey (SF-36) QoL survey and an extensive psychological investigation. RESULTS: Patients with an autograft showed better QoL scales, as compared with mechanical valve recipients. The difference was significant for both the physical (72.72+/ 20.00 vs. 60.27+/-26.07, p = 0.021) and psychological health sum scores (74.71+/ 21.03 vs. 64.71+/-23.49, p = 0.046) and for the subtests of physical functioning (73.72+/-22.44 vs. 62.77+/-25.42, p = 0.049), physical pain (88.39+/-19.13 vs. 73.36+/-27.08, p < or = 0.006), general health perception (64.37+/-17.88 vs. 51.86+/-22.86, p < or = 0.008) and health change (61.89+/-18.94 vs. 50.11+/ 24.37, p = 0.02). The QoL variables did not correlate to pressure gradients, ejection fraction and New York Heart Association functional class. Psychometric tests revealed no meaningful differences between the groups. CONCLUSIONS: This study provides some evidence that patients with pulmonary autografts have greater benefit in terms of QoL, as compared with recipients of mechanical valve substitutes. PMID- 11401140 TI - Influence of ventricular morphology on aerobic exercise capacity in patients after the Fontan operation. AB - OBJECTIVES: This study investigated the influences of ventricular morphology, hemodynamics and clinical findings on exercise capacity in patients after the Fontan operation. BACKGROUND: Determinants of exercise capacity after the Fontan operation remain unclear. METHODS: Peak oxygen uptake (PVo2) was determined in 105 patients by exercise test and compared to hemodynamics and clinical findings. Patients were divided into three groups based on ventricular morphology: those with a right ventricle (group RV), a biventricle (group BV) and a left ventricle (group LV). RESULTS: Ten patients with atrioventricular valve regurgitation (AVVR) or hypoxia exhibited a low PVo2. After excluding these patients, although PVo2 did not correlate with hemodynamics, except ventricular ejection fraction (p < 0.02), it correlated with age at the Fontan operation and exercise test (p < 0.002). The PVo2 was higher in group LV (63+/-9%) than in groups RV (55+/-9%) and BV (55+/-12%) (p < 0.01), while an inverse correlation between PVo2 and age at operation was demonstrated only in group RV (p < 0.05). Groups RV or BV and age at exercise test were associated with a lower PVo2, whereas group LV was an independent predictor of a higher PVo2 (p < 0.01). During 4.2 years of follow-up, a decrease in peak heart rate was related to a decrease in PVo2 (p < 0.05). The PVo2 decreased in group RV (p < 0.01). CONCLUSIONS: In addition to AVVR, hypoxia, and heart rate response, ventricular morphology is related to exercise capacity. Early Fontan operation may be beneficial in terms of exercise capacity, especially in the group RV patients. PMID- 11401141 TI - High-efficiency endovascular gene delivery via therapeutic ultrasound. AB - OBJECTIVES: We studied enhancement of local gene delivery to the arterial wall by using an endovascular catheter ultrasound (US). BACKGROUND: Ultrasound exposure is standard for enhancement of in vitro gene delivery. We postulate that in vivo endovascular applications can be safely developed. METHODS: We used a rabbit model of arterial mechanical overdilation injury. After arterial overdilation, US catheters were introduced in bilateral rabbit femoral arteries and perfused with plasmidor adenovirus-expressing blue fluorescent protein (BFP) or phosphate buffered saline. One side received endovascular US (2 MHz, 50 W/cm2, 16 min), and the contralateral artery did not. RESULTS: Relative to controls, US exposure enhanced BFP expression measured via fluorescence 12-fold for plasmid (1,502.1+/ 927.3 vs. 18,053.9+/-11,612 microm2, p < 0.05) and 19-fold for adenovirus (877.1+/-577.7 vs. 17,213.15+/-3,892 microm2, p < 0.05) while increasing cell death for the adenovirus group only (26+/-5.78% vs. 13+/-2.55%, p < 0.012). CONCLUSIONS: Endovascular US enhanced vascular gene delivery and increased the efficiency of nonviral platforms to levels previously attained only by adenoviral strategies. PMID- 11401142 TI - Cessation of platelet-mediated cyclic canine coronary occlusion after thrombolysis by combining nitric oxide inhalation with phosphodiesterase-5 inhibition. AB - OBJECTIVES: We sought to evaluate the ability of type 5 phosphodiesterase (PDE5) inhibitors to augment the antithrombotic effects of inhaled nitric oxide (NO) in a canine model of platelet-mediated coronary thrombosis after thrombolysis. BACKGROUND: Type 5 phosphodiesterase inhibitors potentiate the ability of NO to inhibit platelet aggregation in vitro by preventing platelet cyclic guanosine monophosphate catabolism. We previously reported that breathing low concentrations of NO gas attenuated, but did not prevent, cyclic flow reductions (CFRs) in a canine model of coronary thrombosis after thrombolysis. METHODS: Cyclic flow reductions were induced after creation of a left anterior descending coronary artery stenosis, endothelial injury, thrombus formation and thrombolysis. Dogs were either untreated or treated with inhaled NO (20 ppm by volume), intravenous zaprinast, intravenous dipyridamole or the combination of inhaled NO with either PDE5 inhibitor (n = 4 per group). RESULTS: Cyclic flow reductions ceased, and complete coronary patency was achieved in all dogs after they breathed NO combined with zaprinast (by 12.0+/-4.7 min [mean +/- SEM]) or dipyridamole (by 9.8+/-4.7 min). The frequency of CFRs was unaffected by NO, dipyridamole or zaprinast alone. Systemic arterial blood pressure and bleeding time were unchanged with any treatment. Ex vivo thrombin-induced platelet aggregation in dogs breathing NO and receiving dipyridamole was reduced by 75+/ 7% (p < 0.05). CONCLUSIONS: The PDE5 inhibitors potentiated the antithrombotic properties of inhaled NO in a canine model of platelet-mediated coronary artery thrombosis after thrombolysis, without prolonging the bleeding time or causing systemic hypotension. PMID- 11401143 TI - Stability of hibernating myocardium in pigs with a chronic left anterior descending coronary artery stenosis: absence of progressive fibrosis in the setting of stable reductions in flow, function and coronary flow reserve. AB - OBJECTIVES: This study was performed to determine whether hibernating myocardium is adaptive or is destined to undergo progressive irreversible injury. BACKGROUND: Previous studies have suggested that hibernating myocardium eventually results in progressive dysfunction. Since serial studies cannot be performed in humans, the temporal progression of physiologic and structural adaptations was evaluated in pigs with hibernating myocardium. METHODS: Pigs were instrumented with a left anterior descending coronary artery (LAD) stenosis (1.5 mm) and underwent physiologic studies three to five months later to quantify regional function, perfusion and 18F-2-deoxyglucose (FDG) uptake. Viability was confirmed by histology and contractile reserve. RESULTS: Hibernating myocardium was characterized by severe regional dysfunction (centerline score, -1.9+/-0.1), reduced resting subendocardial flow (LAD: 0.85+/-0.03 vs. normal: 1.02+/-0.03 ml/min/g, p < 0.01), critically reduced subendocardial flow reserve (adenosine flow: 1.04+/-0.09 ml/min/g, p = NS vs. rest; epinephrine flow: 0.88+/-0.07 ml/min/g, p = NS vs. rest) and increased FDG uptake (0.022+/-0.002 vs. 0.014+/ 0.001 ml/g/min, p < 0.01). Physiologic parameters were not different among animals studied at three (93+/-1 days, n = 27), four (118+/-2 days, n = 26) or five months (150+/-6 days, n = 9). Pathology revealed a small increase in LAD connective tissue (6.4+/-0.4% vs. 4.0+/-0.2%, p < 0.001), with no change over this time frame. CONCLUSIONS: Thus, physiologic and structural features of hibernating myocardium remain constant for at least two months. The absence of functional deterioration or progressive fibrosis suggests that hibernation is adaptive rather than an unstable physiology destined to progress to irreversible injury. The stability of this model appears ideally suited for interventions targeted to improve flow and function in chronically dysfunctional myocardium. PMID- 11401144 TI - Angiotensin-converting enzyme inhibition enhances a subthreshold stimulus to elicit delayed preconditioning in pig myocardium. AB - OBJECTIVES: We assessed the effect of angiotensin-converting enzyme (ACE) inhibition in combination with a subthreshold preconditioning (PC) stimulus to elicit delayed preconditioning against infarction in pig myocardium. BACKGROUND: Bradykinin triggers early PC. Angiotensin-converting enzyme inhibitors increase local bradykinin levels via inhibition of kinin breakdown and have been shown in experimental studies to augment early protection afforded by PC. A role for bradykinin in eliciting delayed PC has not so far been identified. METHODS: We used a two-day protocol. On day 1 (closed chest), pigs were either sham-operated (group 1) or preconditioned, using balloon catheter inflation of the left anterior descending (LAD) coronary artery, with either a full (4 x 5 min PC, group 2) or subthreshold PC stimulus (2 x 2 min PC, group 3). Additional groups were pre-treated with perindoprilat (0.06 mg/kg i.v.) before sham (group 4) or subthreshold PC (group 5). On day 2 (open chest), all pigs were subjected to 40 min occlusion of the LAD followed by 3 h of reperfusion. Infarct size was determined by tetrazolium staining. RESULTS: Group 1 had a mean infarct size of 42.8+/-3.2% of the risk zone. Preconditioning with 4 x 5 min reduced the infarct size to 19.5+/-3.9% (p < 0.05). Groups 3 and 4 had infarct sizes not statistically different from group 1. However, combining perindoprilat with subthreshold PC resulted in a significant limitation of the infarction (18.4+/ 3.1% p < 0.05), comparable with group 2. CONCLUSIONS: This is the first study to show that ACE inhibition can augment a mild ischemic stimulus to induce a protected state 24 h later. PMID- 11401145 TI - Who are the best role models for the fellows? PMID- 11401146 TI - President's page: the Neighborhood Health Watch Program: Save A Victim Everywhere (SAVE). PMID- 11401147 TI - Limitation of vWf meta-analysis in LMWH comparison. PMID- 11401148 TI - Enoxaparin for acute coronary syndromes? PMID- 11401149 TI - Failure of right ventricular recovery of fallot patients after pulmonary valve replacement: delay of reoperation or surgical technique? PMID- 11401150 TI - A real smoker's paradox. PMID- 11401151 TI - Ideal weight, body composition and lipid levels: an unresolved dilemma? PMID- 11401152 TI - Focus on field-portable and miniature mass spectrometers. Presentations from the 12th Sanibel Conference on Mass Spectrometry. PMID- 11401153 TI - Sub-miniature ExB sector-field mass spectrometer. AB - A novel sub-miniature double-focusing sector-field mass spectrometer has been fabricated at the University of Minnesota using a combination of conventional machining methods and thin film patterning techniques typically used in the sensor technology industry. Its design is based on the mass separation capabilities of a 90 degrees cylindrical crossed electric and magnetic sector field analyzer with a 2-cm radius, which under proper conditions is able to effectively cancel the angular and chromatic dispersion of the ion beam, thus improving the resolving power of the instrument. Simulations using finite element analysis and computer modeling were employed to verify and optimize the performance of the proposed instrument before and during its fabrication. The prototype was able to attain a resolving power of 106 full-width at half-maximum (FWHM), a detection limit close to 10 parts per million, a dynamic range of 5 orders of magnitude and a mass range up to 103 Da. Its overall size, including the magnet assembly, is 3.5 cm wide, 6 cm long and 7.5 cm tall, it weighs 0.8 kg, and its power consumption was measured to be 2.5 W. The performance of the instrument was found to be comparable to that of commercial residual gas analyzers, at a fraction of the cost. All these characteristics make this miniature mass spectrometer suitable for portable and low-cost analytical instrumentation. PMID- 11401154 TI - Trade-offs in miniature quadrupole designs. AB - Pressing needs for miniature mass spectrometers became apparent during the last decade in process monitoring and control, space exploration, and environmental screening. Besides the small footprint, common requirements include low cost, low power consumption, field portability, reliability, autonomy, and ease-of-use. Design concepts and construction technologies of miniaturized quadrupole sensors were guided by cost reduction requirements without sacrifice of performance. The first miniature and complete quadrupole mass spectrometer system was introduced as the Micropole sensor. The concept featured a novel technique to assemble and operate multiple miniature quadrupoles in parallel. The short analyzer length offers a significant advantage by enabling direct mass filtering at pressures up in the 10(-2) torr range. High voltages at higher frequencies (10-20 MHz) are required for acceptable mass resolving powers. Additional trade-offs were uncovered in miniature sensors leading to designs optimized for each class of applications. Real time ray tracing of ions injected and filtered in the quadrupole field is used early in the design stage to predict the performance and reliability of the device. PMID- 11401155 TI - Miniaturized EI/Q/oa TOF mass spectrometer. AB - A miniaturized orthogonal time-of-flight mass spectrometer with an electron impact ionization ion source and a rf quadrupole ion guide has been developed. A mass resolving power of m/deltam = 5500 has been obtained in a 0.4 m instrument. The addition of helium at pressures of about 4.0 mtorr into the ion source showed collisional focusing taking place in the rf quadrupole. An automated gas chromatograph designed for air monitoring applications has been coupled to the time-of-flight mass analyzer and tested for the detection of simulants of chemical-warfare agents. PMID- 11401156 TI - Field-portable, high-speed GC/TOFMS. AB - This work is focused on developing a fast gas chromatograph, time-of-flight mass spectrometer (GC/TOFMS) for man-portable field use. The goal is to achieve a total system solution for meeting performance, size, weight, power, cost, and ruggedness requirements for a laboratory in the field. The core technology will also be adaptable to specific applications including real-time point detection for hazardous chemical releases (e.g., chemical weapons), for biological agent signature identification, and for mobile monitoring platforms (e.g., air, ship, truck). Previously we presented results of a feasibility demonstration for a 30 lb field-portable TOFMS system. In this work we present recent progress in integrating a low-power, high-speed GC and show the capability for accurately recording fast GC transients for targeted compound detection using a quadrupole ion trap, time-of-flight instrument (QitTof). PMID- 11401157 TI - Mass spectrometry in the U.S. space program: past, present, and future. AB - Recent years have witnessed significant progress on the miniaturization of mass spectrometers for a variety of field applications. This article describes the development and application of mass spectrometry (MS) instrumentation to support of goals of the U.S. space program. Its main focus is on the two most common space-related applications of MS: studying the composition of planetary atmospheres and monitoring air quality on manned space missions. Both sets of applications present special requirements in terms of analytical performance (sensitivity, selectivity, speed, etc.), logistical considerations (space, weight, and power requirements), and deployment in perhaps the harshest of all possible environments (space). The MS instruments deployed on the Pioneer Venus and Mars Viking Lander missions are reviewed for the purposes of illustrating the unique features of the sample introduction systems, mass analyzers, and vacuum systems, and for presenting their specifications which are impressive even by today's standards. The various approaches for monitoring volatile organic compounds (VOCs) in cabin atmospheres are also reviewed. In the past, ground based GC/MS instruments have been used to identify and quantify VOCs in archival samples collected during the Mercury, Apollo, Skylab, Space Shuttle, and Mir missions. Some of the data from the more recent missions are provided to illustrate the composition data obtained and to underscore the need for instrumentation to perform such monitoring in situ. Lastly, the development of two emerging technologies, Direct Sampling Ion Trap Mass Spectrometry (DSITMS) and GC/Ion Mobility Spectrometry (GC/IMS), will be discussed to illustrate their potential utility for future missions. PMID- 11401158 TI - Underwater mass spectrometers for in situ chemical analysis of the hydrosphere. AB - Underwater mass spectrometry systems can be used for direct in situ detection of volatile organic compounds and dissolved gases in oceans, lakes, rivers and waste water streams. In this work we describe the design and operation of (1) a linear quadrupole mass filter and (2) a quadrupole ion trap mass spectrometer interfaced, in each case, with a membrane introduction/fluid control system and packaged for underwater operation. These mass spectrometry systems can operate autonomously, or under user control via a wireless rf link. Detection limits for each system were determined in the laboratory using pure solutions. The quadrupole mass filter system provides detection limits in the 1-5 ppb range with an upper mass limit of 100 amu. Its power requirement is approximately 95 Watts. The ion trap system has detection limits well below 1 ppb, an upper mass limit of 650 amu and MS/MS capability. Its power consumption is on the order of 150 Watts. The present membrane limits analysis to non-polar compounds (<300 amu) with analysis cycles of 5-15 minutes. Deployments of both types of instruments are described, along with a discussion of the challenges associated with in-water mass spectrometry and descriptions of alternative in-water mass spectrometer configurations. PMID- 11401159 TI - Environmental and forensic applications of field-portable GC-MS: an overview. AB - GC-MS can provide analytical information that is most reliable for many types of organic analyses. As field-portable GC-MS analytical systems evolve, the application scenarios have diversified as well. With the development of rugged fieldable systems, these instruments were demonstrated to be usable in the harsh environment of the jungle and in chemical demilitarization or military reconnaissance situations. Continuous unattended operations of a GC-MS for 12- or 24-hour monitoring applications in the field have been shown to be possible. A real-time algorithm strategy is proposed, which can be developed to aid in the advancement of field-portable mass spectrometry applied to chemical warfare agent analysis in military vehicles and can be used to raise the standard for field data quality. Each of these capabilities is discussed with the intent on reviewing analysis situations that can be expanded because of developments in field GC-MS instrumentation. PMID- 11401160 TI - Reactions of O*- with methyl benzoate: a negative ion chemical ionization and Fourier transform ion cyclotron resonance study. AB - The reactions of O*- with methyl benzoate have been examined by the measurement of negative ion chemical ionization (NICI) mass spectra using a CI source, with confirmatory studies carried out on a Fourier transform ion cyclotron resonance mass spectrometer. Reaction mechanisms have been elucidated using isotopically labeled esters. Nucleophilic attack at the carbonyl carbon and the aromatic ring were important reaction pathways. Nucleophilic attack at the carbonyl carbon was followed by the production of products (C6HsCO2- and CH3OCO2-) characteristic of radical, beta-fragmentation. Using 18O-labeled methyl benzoate, the SN2 reaction was found to account for a smaller percentage, 21(+/-1)%, of the benzoate product. Aromatic ring attack resulted in formation of [M + O - H]- and [M - 2H]* ions. Although aryl hydrogens accounted for most H2*+ abstracted by O*-, evidence for abstraction of HarylH*+alkyl and HalkylH*+alkyl was also found. Although present at much lower abundance, dehydrobenzoate, dehydrophenoxy, and C7H6*- ([M - 2H - CO2]*-) radical anions were also observed. An Haryl/Halkyl exchange associated with formation of the benzoate anion was attributed to an Halkyl abstraction that occurred within the methanol/dehydrobenzoate ion-dipole complex. The [M - 2H]*-, dehydrobenzoate, dehydrophenoxy, and [M - 2H - CO2]*- ion signals were quenched by reaction with O2. Conditions required for production of O*- spectra under NICI conditions were also examined. PMID- 11401161 TI - Determination of flavone, flavonol, and flavanone aglycones by negative ion liquid chromatography electrospray ion trap mass spectrometry. AB - Eleven naturally occurring flavonoid aglycones, belonging to the representative flavone, flavonol, and flavanone types were separated by high performance liquid chromatography and analyzed on-line with negative ion electrospray ionization tandem mass spectrometry (ESI-MS/MS). In order to resolve the MS/MS spectra obtained, each compound was reinvestigated by direct loop injections using an ion trap mass spectrometer. The MSn spectra obtained allowed us to propose plausible schemes for their fragmentation supported by the analysis of five complementary synthetic flavonoid aglycones. The negative ion ESI-MS/MS behavior of the different aglycones investigated in this study revealed interesting differences when compared with the previously described patterns obtained using various ionization techniques in positive ion. Thus, concerning the retro Diels-Alder (RDA) fragmentation pathways, several structurally informative anions appeared highly specific of the negative ion mode. In addition, a new lactone-type structure, instead of a ketene, was proposed for a classic RDA diagnostic ion. We also observed unusual CO, CO2, and C3O2 losses which appear to be characteristic of the negative ion mode. All these results and these unusual neutral losses show that the negative ion mode was a powerful complementary tool of the positive ion mode for the structural characterization of flavonoid aglycones by ESI-MS/MS. PMID- 11401162 TI - Protein fluorescence measurements within electrospray droplets. AB - The conformation of cytochrome c molecules within electrospray droplets is investigated by monitoring the laser induced fluorescence of its single tryptophan residue (Trp-59). By increasing the alcohol concentration of the electrosprayed solutions, protein denaturation is induced, giving rise to significant changes in the intensity of the detected fluorescence. Comparison with analogous denaturation experiments in solution provides information about the relative protein conformations and differences between the bulk-solution and droplet environments. Both electrospray-plume and bulk-solution fluorescence measurements using low methanol concentration solutions indicate the presence of folded protein structures. At high methanol content, fluorescence measurements are consistent with the presence of partly denatured or unfolded conformations. At intermediate methanol content, differences are observed between the extent of denaturation in solution and that within the droplets, suggesting electrosprayed proteins have more compact structures than those detected in bulk measurements using solutions of similar composition. This infers that some fraction of the proteins within the droplets have refolded relative to their bulk-solution conformation. Protein denaturation experiments using the low vapor pressure solvent 1-propanol indicate that differences between the droplet and solution measurements are not due to solvent evaporation effects. It is suggested that different droplet conformations are more likely the result of protein diffusion to the droplet surface and effects of the droplet/air interface. To our knowledge, these are the first reported measurements of protein fluorescence within electrospray droplets. PMID- 11401163 TI - The mechanism of matrix to analyte proton transfer in clusters of 2,5 dihydroxybenzoic acid and the tripeptide VPL. AB - Intracluster proton transfer from the matrix-assisted laser desorption/ionization matrix 2,5-dihydroxybenzoic acid (DHB) to the peptide valyl-prolyl-leucine has been investigated as a function of excitation laser wavelength and power. Ionization laser power studies at 308 nm indicate that cluster ionization occurs with a two-photon dependence, whereas matrix-to-analyte proton transfer and cluster dissociation requires an additional photon. At 266 nm, two-photon absorption leads to both cluster ionization and cluster dissociation/proton transfer. A consideration of these results clearly indicates that analyte protonation occurs following ionization of the cluster to produce a radical cation matrix/analyte cluster. Mass spectral features also indicate that mixed DHB/peptide cluster ionization can occur via two-photon ionization at wavelengths as long as 355 nm. These results suggest a reduction in the ionization potential of larger mixed DHB/peptide clusters of greater than 1 eV. The reduced ionization potential seen in these clusters suggests that radical cation initiated proton transfer remains a viable mechanism for analyte protonation in matrix-assisted laser desorption/ionization at these longer wavelengths. PMID- 11401164 TI - Silver cluster interferences in matrix-assisted laser desorption/ionization (MALDI) mass spectrometry of nonpolar polymers. AB - Potential difficulties associated with background silver salt clusters during matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) of nonpolar polymers are reported. Silver salt cluster ions were observed from m/z 1500 to 7000 when acidic, polar matrices, such as 2,5-dihydroxybenzoic acid (DHB), all-trans-retinoic acid (RTA) or 2-(4-hydroxyphenylazo)benzoic acid (HABA), were used for the analysis of nonpolar polymers. These background signals could be greatly reduced or eliminated by the use of nonpolar matrices such as anthracene or pyrene. Representative examples of these background interferences are demonstrated during the analysis of low molecular weight nonpolar polymers including polybutadiene and polystyrene. Nonpolar polymers analyzed with acidic, polar matrices (e.g., RTA) and silver cationization reagents can yield lower quality mass spectral results when interferences due to silver clusters are present. Replacing the polar matrices with nonpolar matrices or the silver salts with copper salts substantially improved the quality of the analytical results. In addition, it was found that silver contamination cannot be completely removed from standard stainless steel sample plates, although the presence of silver contamination was greatly reduced after thorough cleaning of the sample plate with aluminum oxide grit. Carry-over silver may cationize polymer samples and complicate the interpretation of data obtained using nonpolar matrices in the absence of added cationization reagents. PMID- 11401165 TI - Elucidation of the initial step of oligonucleotide fragmentation in matrix assisted laser desorption/ionization using modified nucleic acids. AB - To probe the mechanism of gas-phase oligonucleotide ion fragmentation, modified oligonucleotides were studied using matrix-assisted laser desorption/ionization. The oligonucleotides were of the form 5'-TTTTXTTTTT, where X was a modified nucleotide. Modifications included substitution of hydroxy, methoxy, amino, and allyl groups at the 2'-position of the deoxyribose. The modified ribose contained adenine, guanine, cytosine, or uracil bases. For comparison, we studied oligomers where X was an unmodified adenosine, guanosine, cytidine, thymidine, or uridine deoxyribonucleotide. We found a very strong dependence of the matrix-to-analyte ratio on fragmentation for these oligomers. Analysis of these modifications suggests that the initial fragmentation step in MALDI-MS involves a two-step (E1) elimination of the base. PMID- 11401166 TI - Elimination of the concentration dependence in mass isotopomer abundance mass spectrometry of methyl palmitate using metastable atom bombardment. AB - An important problem in mass isotopomer abundance mass spectrometry (MIAMS) is the dependence of measured mass isotopomer abundances on sample concentration. We have evaluated the role of ionization energy on mass isotopomer abundance ratios of methyl palmitate as a function of sample concentration. Ionization energy was varied using electron impact ionization (EI) and metastable atom bombardment (MAB). The latter generates a beam of metastable species capable of ionizing analyte molecules by Penning ionization. We observed that ionization of methyl palmitate by EI (70 eV) showed the greatest molecular ion fragmentation and also showed the greatest dependence of relative isotopomer abundance ratios on sample concentration. Ionization using the 3P2 and 3P0 states of metastable krypton (9.92 and 10.56 eV, respectively) resulted in almost no molecular ion fragmentation, and the isotopomer abundances quantified were essentially independent of sample concentration. Ionization using the 3P2 and 3P0 states of metastable argon (11.55 and 11.72 eV, respectively) showed molecular ion fragmentation intermediate between that of EI and MAB(Kr) and showed an isotopomer concentration dependence which was less severe than that observed with EI but more severe than that observed with MAB(Kr). The observed decrease in the dependence of isotopomer abundance on sample concentration with a decrease in molecular ion fragmentation is consistent with the hypothesis that proton transfer from a fragment cation to a neutral molecule is the gas phase reaction mechanism responsible for the concentration dependence. Alternative explanations, e.g., hydrogen abstraction from a neutral molecule to a molecular cation, is not supported by these results. Moreover, the MAB ionization technique shows potential for eliminating one source of error in MIAMS measurements of methyl palmitate, in particular, and of fatty acids methyl esters, in general. PMID- 11401167 TI - Biodegradable interference screw augmentation reduces tunnel expansion after ACL reconstruction. AB - This study assessed 40 patients who underwent anterior cruciate ligament reconstruction with hamstring autografts using our standard technique. Twenty patients underwent tibial fixation augmentation with a biodegradable interference screw, and the remaining 20 patients did not. Patient groups were compared using radiographic evaluation of bone tunnel diameter, physical examination, and arthrometer measurements. The two groups differed in only one way: the placement of the tibial biodegradable interference screw. Biodegradable interference screw augmentation resulted in decreased distal tibial and some femoral tunnel diameters at both 3- and 12-month follow-up. There was no significant difference in the Lachman and arthrometer testing in either group of patients at the termination of this study. These results indicate that although distal tibia tunnel diameter reduction is significant, it does not appear to have any clinical significance in the first year. PMID- 11401168 TI - Retrograde femoral nailing: a focus on the knee. AB - A consecutive series of 23 patients with reamed retrograde femoral nails was reviewed. Nails were placed through the intercondylar notch with a minimal incision. Nineteen patients with retrograde femoral nails were available at an average follow-up of 19.3 months. The union rate was 100% with no infections or malunions. No second surgeries were required for union. Knee range of motion averaged 109 degrees and was greater in those patients with shaft fractures (117 degrees) than in those with supracondylar-intercondylar fractures (91.3 degrees) (P=.02). Pain (0-3 scale) averaged 0.36. Hospital for Special Surgery knee scores averaged 80.4 (90% good or excellent results). Minor knee pain (55%) and secondary surgeries (35%) were common. The only fair or poor results were in patients with preexisting osteoarthritis. A literature review of 14 papers and abstracts was conducted. Exposure, often extensive initially, is more recently percutaneous. The infection rate is acceptable (0-14%), with knee sepsis uncommon. Lower union rates were observed for supracondylar femur fractures (80% 84%) than for femoral shaft fractures (85%-100%) after a single surgery. Second surgeries are common (14%-60%). Varus/valgus malunion, common (12%-29%) with the initial extrarticular entry site, occurs less with the intercondylar entry site. The antegrade femoral nail allows for better control of proximal shaft fractures, while the retrograde femoral nail is more reliable in controlling distal shaft fractures. Rotational malunion still remains a problem. Mild knee pain is common (13%-60%). The treatment of supracondylar femur nonunions with retrograde femoral nailing is disappointing. PMID- 11401169 TI - Spontaneous repair of infrapatellar fat pad after resection of para-articular chondroma occupying infrapatellar space. PMID- 11401170 TI - Subcutaneous migration of bioabsorbable meniscal arrows. PMID- 11401171 TI - Fractures of the tibial plateau. PMID- 11401172 TI - Operative management of fractures of the tibial plateau. PMID- 11401173 TI - Hybrid external fixation for tibial plateau fractures. PMID- 11401174 TI - Component size asymmetry in bilateral total knee arthroplasty. AB - A review of 268 consecutive patients undergoing bilateral total knee arthroplasty (TKA) was performed to determine whether component size asymmetry exists in patients undergoing bilateral TKAs. Component sizes were selected based on preoperative radiographic templating and intraoperative sizing measurements irrespective of the component sizes chosen for the other knee. All radiographs were evaluated according to described criteria. Component sizes used for the femur, tibia, and patella were compared between the right and left knees. Of the 268 bilateral TKAs, 18 (6.7%) femoral components varied in size between right and left knees. There were no statistical differences for patellar or tibial component size asymmetry or knee function pre- or postoperatively. Patients with asymmetrically sized femoral components had no statistical difference between left and right knees with respect to knee score, pain, function, range of motion, incidence of lateral release, or complications. PMID- 11401175 TI - Current practices and opinions in ACL reconstruction and rehabilitation: results of a survey of the American Orthopaedic Society for Sports Medicine. AB - Many different surgical techniques and rehabilitation protocols have evolved for the treatment of anterior cruciate ligament (ACL) injuries, and there is a lack of agreement as to which approach results in the best outcome. Members of the American Orthopaedic Society for Sports Medicine (AOSSM) were surveyed to determine their current ACL reconstruction technique and opinions regarding preoperative and postoperative management. In 1999, members of the AOSSM were mailed surveys asking about their current treatment of ACL injuries. Approximately 76% of the active members responded to the survey, of which a large percentage (92%) currently performs ACL reconstructions. Both the experience of the surgeon and annual number of ACL reconstructions performed were recorded. Most responding surgeons routinely perform ACL reconstructions 3-6 weeks following an acute ACL injury using an endoscopic technique. Bone-patellar tendon bone (BPTB) with interference screw fixation was the technique of choice for most respondents, with the majority performed on an outpatient basis. Rehabilitation protocols showed more variation regarding postoperative weight bearing, immobilization and bracing, length of physical therapy, and return to sport. Most surgeons prefer early postoperative full weight bearing with an average of 3.8 weeks of postoperative bracing. Physical therapy typically ranged from 1-4 months with return to sport at 6-7 months, generally with a functional brace. Patients with BPTB reconstruction were allowed the earliest return to full activity. Although previous clinical and biomechanical studies show good-excellent results with different ACL reconstruction and rehabilitation techniques, currently most surgeons practicing as members of the AOSSM continue to prefer BPTB grafts with metal interference screw fixation. However, there is less consensus regarding the specific postoperative rehabilitation protocol. PMID- 11401176 TI - The relationship of femoral intercondylar notch width to height, weight, and sex in patients with intact anterior cruciate ligaments. AB - Intercondylar notch width, femoral bicondylar width, height, and weight of patients with intact anterior cruciate ligaments were measured to determine whether intercondylar notch width was related to body size. A 45 degrees weight bearing posteroanterior radiograph was obtained for 315 men and 163 women. Notch width and bicondylar width was measured at one-half notch height. Mean notch width for men was statistically significantly wider than for women (17.1 mm versus 14.7 mm, respectively; P<.01). There was no statistically significant correlation between height and notch width for men (r=-0.0019; P=.97) or women (r=0.1308; P=.10). No significant correlation existed between weight and notch width for men (r=-0.0311; P=.58) or women (r=0.0523; P=.51). Analysis of variance showed height and weight were not significant covariates in notch width for either men (P=.44) or women (P=.91). Women of the same height and weight as men had significantly narrower notches (P<.01). There was a statistically significant correlation between wider femoral bicondylar widths and higher weight for men (r=0.694; P<.01) and women (r=0.821; P<.01). Similarly, there was a statistically significant correlation between wider femoral bicondylar widths and increased height for men (r=0.670; P<.01) and women (r=0.785; P<.01). These data demonstrate height and weight are poor predictors of intercondylar notch width. Therefore, one cannot assume body size is a predictor of notch width. Furthermore, because mean notch width does not increase with increasing height and weight, the notch width index calculation cannot accurately reflect the size of the intercondylar notch. PMID- 11401177 TI - Reliability of the quadriceps angle measurement. AB - The quadriceps angle (Q-angle) is used to determine patellofemoral alignment. Although this measurement has been used to evaluate and treat patellofemoral joint pathology, few studies have examined its reliability. This study evaluated the interobserver and intraobserver reliability of the Q-angle measurement. To investigate the interobserver reliability of the Q-angle, 25 individuals of varying levels of training served as observers and participants as each measured the other 24 participants. To investigate the intraobserver reliability of the Q angle, 3 of the observers measured 13 of the participants an additional 2 times. Additionally, clinically derived Q-angle measurements were compared with radiographically derived measurements. The reliability analysis was performed using intraclass correlation coefficients. For interobserver measurements, the intraclass correlation coefficients ranged from 0.17-0.29 for the four variables evaluated (right and left, extension and flexion). For intraobserver measurements, the intraclass correlation coefficients ranged from 0.14-0.37. The average intraclass correlation coefficient between the clinically and radiographically derived measurements ranged from 0.13-0.32. This study demonstrates poor interobserver and intraobserver reliability of Q-angle measurement and poor correlation between clinically and radiographically derived Q-angles. PMID- 11401178 TI - Bioavailability of the oral selective phosphodiesterase 4 inhibitor cilomilast. AB - STUDY OBJECTIVE: To determine the absolute bioavailability of cilomilast, and assess the effects of food, dosing time, and coadministration of antacid agents on its bioavailability and pharmacokinetics in healthy volunteers. SETTING: Clinical pharmacology unit. DESIGN: Five prospective pharmacokinetic studies: one single-blind, dose-escalating, placebo-controlled trial; four open-label, randomized studies. SUBJECTS: Ninety-six healthy adult volunteers who were nonsmokers. INTERVENTION: In the first study, four subjects received intravenous cilomilast 1, 2, and 4 mg. In the second study, 16 subjects received oral cilomilast 15 mg or intravenous cilomilast 4 mg. In the other three studies, a total of 76 subjects were given single oral 15-mg doses; one study compared its effects in fed versus fasted subjects, one looked for differences of morning versus evening dosing, and one examined coadministration with aluminum hydroxide magnesium hydroxide. MEASUREMENTS AND MAIN RESULTS: After intravenous administration of cilomilast, plasma concentrations increased in an approximately dose-proportional manner; the half-life, approximately 6.5 hours, was dose independent. Cilomilast clearance and volume of distribution were small. After oral dosing, the absolute bioavailability was consistently close to 100%. Absorption was slower in fed subjects than in fasted (median 2-hr delay in time to reach maximum plasma concentration, average 39% reduction in maximum plasma concentration), but the area under the concentration-time curve from time zero to infinity (systemic availability) was unaffected. Pharmacokinetic parameters were not influenced by time of dosing or coadministration of antacid. CONCLUSION: The absolute bioavailability of oral cilomilast was 100%; it was not adversely affected by time of dosing or coadministration with food or antacid. PMID- 11401179 TI - Hyperbilirubinemia during quinupristin-dalfopristin therapy in liver transplant recipients: correlation with available liver biopsy results. AB - STUDY OBJECTIVE: To review the liver histopathology in transplant recipients who developed hyperbilirubinemia during therapy with quinupristin-dalfopristin, a new streptogramin antibiotic, and to ascertain whether objective histologic evidence of adverse drug effect could be correlated to serum bilirubin levels. DESIGN: Retrospective analysis. SETTING: University of Pittsburgh Medical Center. PATIENTS: From a database of 34 liver recipients who received quinupristin dalfopristin for vancomycin-resistant Enterococcus faecium infection who were prospectively enrolled in a multicenter, open-label, emergency-use protocol, the data for a subset of 25 patients who underwent one or more liver biopsies during therapy were reviewed for this study. INTERVENTIONS: Quinupristin-dalfopristin was administered intravenously at 7.5 mg/kg every 8 hours. Available serum bilirubin levels from before, during, and 1 week after therapy were tabulated. Liver biopsy results obtained within 1 week before and during therapy were retrospectively reviewed. Histopathologic results were characterized and correlated to bilirubin level. MEASUREMENTS AND MAIN RESULTS: Cholestatic changes were already present in 15 of 17 patients who underwent biopsy before therapy. During therapy, the most common findings from 40 biopsies (25 patients) were cholestasis (33 biopsies), acute rejection (10), and periportal inflammation (8). There was no evidence of drug-specific histopathologic injury. CONCLUSION: Hyperbilirubinemia in these patients was likely multifactorial and most frequently due to sepsis and prior graft injury. PMID- 11401180 TI - Active transport of nitrofurantoin into human milk. AB - STUDY OBJECTIVE: To determine the extent to which nitrofurantoin is transferred into human milk. DESIGN: Prospective, single-dose pharmacokinetic study. SETTING: University-affiliated clinical research center. PATIENTS: Four healthy lactating women 8-26 weeks postpartum. INTERVENTION: All subjects received a single, oral, 100-mg dose of nitrofurantoin macrocrystals with food. Serial serum and milk samples were obtained and analyzed by high-performance liquid chromatography. MEASUREMENTS AND MAIN RESULTS: Milk pH, milk fat partitioning, and protein binding in serum and milk were determined. Predicted milk:serum ratio (M:S) was compared with the observed M:S. Nitrofurantoin M:S predicted was 0.28+/-0.05, whereas M:S observed was 6.21+/-2.71. Average milk concentration was 1.3 mg/L, and estimated suckling infant dosage was 0.2 mg/kg/day or 6% of maternal dose (mg/kg). CONCLUSIONS: Nitrofurantoin is actively transported into human milk, achieving concentrations in milk greatly exceeding those in serum. Concern is warranted for suckling infants younger than 1 month old, or for infants with a high frequency of glucose-6-phosphate dehydrogenase deficiency or sensitivity to nitrofurantoin. PMID- 11401181 TI - Drug therapy and prevalence of erectile dysfunction in the Massachusetts Male Aging Study cohort. AB - STUDY OBJECTIVE: To examine the association of commonly used drugs with erectile dysfunction (ED) at two time points. DESIGN: Population-based, cross-sectional, survey analysis. PARTICIPANTS: Randomly selected cohort of men in the Massachusetts Male Aging Study (MMAS) that included 1476 men for the baseline (1987-1989) and 922 for the follow-up (1995-1997) analyses. INTERVENTION: Crude associations between specific drug categories were examined with chi2 statistics. Logistic regression analysis was used to separate the effect of drugs from the influence of heart disease, hypertension, untreated diabetes, or depressive symptoms. MEASUREMENTS AND MAIN RESULTS: In the MMAS, medical history, current drug use, and erectile function status were ascertained with in-home interviews. In unadjusted analyses, thiazide and nonthiazide diuretics, beta-blockers, calcium channel blockers, angiotensin-converting enzyme inhibitors, benzodiazepines, digitalis, nitrates, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, and histamine2 receptor antagonists were associated with prevalent ED. Adjustment for comorbidities and health behaviors attenuated these associations, with only nonthiazide diuretics and benzodiazepines remaining statistically significant. CONCLUSION: Several common drugs may increase prevalence of ED; however, additional data from larger populations are needed to determine whether these associations are independent of underlying health conditions and to explore the effects of dosage and duration of use. PMID- 11401182 TI - Economic impact of neutropenia and febrile neutropenia in breast cancer: estimates from two national databases. AB - STUDY OBJECTIVE: To estimate the economic burden of neutropenia and febrile neutropenia in female breast cancer hospital admissions in the United States. DESIGN: Retrospective database analysis. PATIENTS: Female admissions with a breast cancer diagnosis. MEASUREMENTS AND MAIN RESULTS: By reviewing two national databases (Healthcare Costs and Utilization Project, MarketScan), length of stay and charge or payment/admission were estimated from 1994-1996. Neutropenic and febrile neutropenic admissions were longer and incurred higher charges and payments than nonneutropenic and afebrile neutropenic admissions, respectively (p<0.05). The difference in mean charges between neutropenic and nonneutropenic admissions decreased from $13,143 in 1994 to $6913 in 1996, whereas the difference in payment was $4957 (adjusted to 1996 dollars). The difference in mean charges between febrile and afebrile neutropenic admissions decreased from $11,570 in 1994 to $2873 in 1996, whereas the difference in payment was $2390 (adjusted to 1996 dollars). CONCLUSION: There was a trend toward decreased charges for inpatient admissions with neutropenia in patients with breast cancer (1994-1996). Interventions that reduce the frequency of neutropenia and febrile neutropenia could reduce hospitalization costs of breast cancer admissions. PMID- 11401183 TI - Update on strategies to improve thrombolysis for acute myocardial infarction. AB - Therapy for acute myocardial infarction involves rapid restoration of blood flow through a coronary artery that has been occluded by a ruptured atherosclerotic plaque. Thrombolytic therapy, the pharmacologic standard to achieve this outcome, significantly improves survival; however, current regimens have limitations: they can fail to achieve complete reperfusion, they can cause significant bleeding events, and they can result in reocclusion. In addition, complex regimens of some agents can cause dosing errors. Accordingly, newer compounds were developed to address some of these issues, and alternative strategies are being implemented. The combination of platelet glycoprotein IIb-IIIa receptor inhibitors plus thrombolytic agents produced promising results in clinical trials, including faster clot lysis and greater flow rates than either therapy alone. The addition of unfractionated heparin or low-molecular-weight heparin to thrombolytic antiplatelet therapy is being evaluated, as is administration of thrombolytic antiplatelet before percutaneous coronary intervention. PMID- 11401184 TI - Ziprasidone, a new atypical antipsychotic drug. AB - Although the introduction of antipsychotic drugs in 1954 was a breakthrough in the treatment of patients with schizophrenia, these agents have a number of adverse effects that limit effectiveness and compliance. The atypical antipsychotic drugs provide an improved tolerability profile, particularly in minimizing extrapyramidal side effects; however, they are associated with significant weight gain, which may be related to growing evidence linking the atypical agents with diabetes and hyperlipidemia. Ziprasidone, a new atypical antipsychotic drug, was demonstrated in clinical trials to be more efficacious than placebo and similar in efficacy to haloperidol in the treatment of schizophrenia. Like the existing atypical agents, ziprasidone has a rate of extrapyramidal side effects similar to that of placebo and does not cause significant elevations in prolactin levels. In contrast, ziprasidone has a low propensity for causing weight gain. For patients requiring an antipsychotic drug, ziprasidone represents a new treatment option with a limited adverse effect profile. PMID- 11401185 TI - Practice-based research: lessons from community pharmacist participants. AB - We designed this project to determine community pharmacists' opinions regarding the challenges and motivations of their recent participation in a pharmacy practice-based research study At the conclusion of a randomized, multicenter study, 87 community pharmacist-investigators were sent a questionnaire that explored four areas: motivating factors to participate, barriers to participation, communication tools used by study coordinators, and design issues for future studies. Fifty-eight (67%) completed questionnaires were returned. Key factors motivating participation in the study were desire to improve the profession and opportunity to learn. Time was the greatest barrier to participation. Pharmacy practice-based research has two distinct advantages. First, it translates clinical knowledge into direct application in the community. Second, it provides needed data to demonstrate the value of enhanced pharmacy practice. Thorough understanding of pharmacists' opinions is necessary to optimize the design of future studies. PMID- 11401186 TI - Implementation and evaluation of guidelines for use of enoxaparin as deep vein thrombosis prophylaxis after major trauma. AB - Although enoxaparin is more efficacious than many other deep vein thrombosis (DVT) prevention strategies after trauma, its routine use in trauma patients at low risk for venous thrombosis is unlikely to be cost-effective and may be deleterious if risk factors for bleeding are present. By way of consensus of opinion of trauma surgeons and pharmacists, enoxaparin DVT prophylaxis guidelines were developed, implemented, and evaluated. Fifty patients with major orthopedic or spinal trauma were followed throughout hospitalization. Enoxaparin use and frequency of DVT, pulmonary embolism (PE), thrombocytopenia, and enoxaparin related major bleeding (overt bleeding associated with a hemoglobin decrease > or = 2 g/dl, need for > or = 2 units of packed red blood cells, or need for surgery) were recorded. All pharmacist interventions pertaining to enoxaparin prophylaxis were collected. Average patient age was 45.6+/-19.5 years, average Injury Severity Score was 19.0+/-11.2, and average length of hospitalization was 14.3+/ 10.0 days. Most injuries were related to motor vehicles (52%) and falls (30%). Sites of injury were femur or tibia (52%), pelvis or acetabulum (32%), hip (20%), and spinal cord (12%). Two-thirds (72%) of patients received enoxaparin during part of their hospital stay (on average, for 53% of the duration of hospitalization). Sequential compression devices and vena caval filters were used in 86% and 10% of patients, respectively. Duplex-proven DVT occurred in two patients, and angiography-proven PE developed in one patient. Enoxaparin-related major bleeding and thrombocytopenia occurred in three and one patient(s), respectively. Pharmacists recommended enoxaparin initiation in nine (18%) patients and discontinuation of the agent in seven (14%) patients (one for bleeding; six for lack of indication). Most recommendations (78%) were accepted. Data from the 50 patients in this study showed fewer thrombotic complications but more bleeding than the frequencies found in controlled studies. It is unclear whether the large number of days that patients did not receive enoxaparin was due to fears of enoxaparin-related bleeding or other factors. PMID- 11401187 TI - Cerebrospinal fluid concentrations of quinupristin-dalfopristin in a patient with vancomycin-resistant Enterococcus faecium [correction of faecalis] ventriculitis. AB - A 44-year-old man was treated successfully for vancomycin-resistant Enterococcus faecium (VREF) ventriculitis with intrathecal quinupristin-dalfopristin 1 mg, 2 mg, and 4 mg, and other intravenous antibiotics. Cerebrospinal fluid samples were collected before and after the 1-mg and 2-mg doses to determine the concentrations of quinupristin-dalfopristin and its active metabolites. Concentrations were above the minimum inhibitory concentration for VREF immediately after unclamping the extraventricular drain and were quantifiable for at least 7 hours. PMID- 11401188 TI - Pegylated liposomal doxorubicin: tolerability and toxicity. AB - We evaluated the tolerability and toxicity attributed to pegylated liposomal doxorubicin (PL-DOX) in women with recurrent or refractory ovarian cancer, and reviewed procedures to prevent or treat toxicity induced by the agent. Medical records of 13 women who received PL-DOX between October 1997 and December 2000 were reviewed. Patients 1-8 received PL-DOX once it was added to the hospital formulary in 1997. Patients 9-13 received it after medical staff education. Data on premedications, number of cycles, dosage, length of infusion, tolerability, side effects, and indicators for response were collected. The median number of cycles and cumulative dose/patient of PL-DOX were higher (6 and 420 mg) in the second group than in the first group (2 and 240 mg). Patient factors such as duration of disease and number of chemotherapy cycles influenced tolerability. One patient experienced a life-threatening adverse reaction within minutes of receiving the first dose. Treatment was discontinued, and she was resuscitated successfully. Other dose- or treatment-limiting complications (neutropenia, stomatitis, plantar-palmar erythrodysesthesia) were documented. Toxicity management consisted of dosage reduction or treatment delay; treatment often was discontinued. Patients with recent disease tolerated more cycles of PL-DOX when given early in recurrence compared with heavily pretreated women with long standing disease. Tolerability was not necessarily indicative of response. The agent is simple to administer, but its tolerability and lack of uniform toxicity management remain concerns. PMID- 11401189 TI - Fatal hemolysis after high-dose etoposide: is benzyl alcohol to blame? AB - A 53-year-old African-American man with relapsed non-Hodgkin's lymphoma developed seizures and respiratory arrest 2 hours after an infusion of high-dose etoposide in preparation for an autologous bone marrow transplant. Laboratory tests revealed both rapid hemolysis and severe metabolic acidosis. The patient died the following day. Based on toxicities observed, we suspect that our patient possessed an ethnic polymorphism of the enzyme alcohol dehydrogenase. Further research is required to determine the relationship between the benzyl alcohol metabolic rate and toxicity and genetic polymorphisms of alcohol dehydrogenase in African-Americans. PMID- 11401190 TI - Simvastatin-associated memory loss. AB - The statins are widely used to treat dyslipidemias. They are generally associated with mild adverse effects, but rarely, more serious reactions may occur. A 51 year-old man experienced delayed-onset, progressive memory loss while receiving simvastatin for hypercholesterolemia. His therapy was switched to pravastatin, and memory loss resolved gradually over the next month, with no recurrence of the adverse effect. PMID- 11401191 TI - Adulteration: its various meanings. PMID- 11401193 TI - New developments in the prevention and treatment of venous thromboembolism. AB - Although unfractionated heparin has been widely used for preventing and treating venous thromboembolism, low-molecular-weight heparins (LMWHs) have been extensively studied. In particular, LMWHs have been valuable in the prevention of venous thromboembolism in high-risk surgical patients, such as those undergoing total joint replacement, and in high-risk medical patients. Recent studies indicated that the extended use of LMWHs after discharge in patients undergoing total hip replacement significantly decreases the frequency of venous thrombosis compared with placebo. Furthermore, LMWHs have been shown to be as effective and safe as unfractionated heparin for the initial treatment of both deep vein thrombosis and pulmonary embolism and have extended the treatment of these conditions into the outpatient setting. New recommendations from the sixth American College of Chest Physicians Consensus Conference on Antithrombotic Therapy and the rationale for change are discussed. PMID- 11401194 TI - Low-molecular-weight heparins are essentially the same for treatment and prevention of venous thromboembolism. AB - The pharmacodynamic properties of low-molecular-weight heparins differ. Data from randomized clinical trials show that despite these differences, the agents have similar efficacy and safety profiles in preventing and treating new and recurrent venous thromboembolism in patients who underwent general surgery or total hip or knee replacement. Dalteparin, enoxaparin, or tinzaparin, when administered at the dosages used in the reviewed clinical trials, are essentially indistinguishable. PMID- 11401195 TI - Differentiation of the low-molecular-weight heparins. AB - The three low-molecular-weight heparins (LMWHs) available in the United States have been extensively evaluated for a wide array of indications. Properties associated with one LMWH cannot be assumed to be the same as those associated with another LMWH, as they are different pharmacologic entities. Therefore, therapeutic interchange of these agents is inappropriate. The pharmacokinetic and pharmacodynamic differences among LMWHs can be explained by comparing methods of preparation, molecular structures, half-lives, antithrombin- and non-antithrombin mediated actions, effect on thrombus, and dosing interval. The Food and Drug Administration-approved indications and their respective levels of clinical evidence further differentiate these agents. A dichotomy in the results of clinical trials has been observed with the LMWHs. As the LMWHs are distinct compounds that each possess unique pharmacokinetic and pharmacodynamic profiles, treatment decisions should be based on the available safety and efficacy data for each LMWH. Agents should be prescribed only for those indications for which they have been shown to be effective and only at dosages that have been studied. PMID- 11401196 TI - Chemorepellent signaling through the PACAP/lysozyme receptor is mediated through cAMP and PKC in Tetrahymena thermophila. AB - Pituitary adenylate cyclase-activating peptide and lysozyme are potent chemorepellents which act through the same receptor in Tetrahymena. Using in vivo behavioral studies, we have found that the pituitary adenylate cyclase-activating peptide/lysozyme receptor appears to signal through a G-protein pathway which is mediated through both adenosine 3'5'monophosphate and protein kinase C. Avoidance to pituitary adenylate cyclase-activating peptide and lysozyme is inhibited by the G-protein inhibitor, guanosine 5'-O-(2thiodiphosphate), the adenosine 3'5'monophate analog, Rp-adenosine-3', 5' cyclic monophosphorothioate, and the protein kinase C inhibitors, calphostin C and bisindolylmaleimide IV. A proposed model for signaling through the pituitary adenylate cyclase-activating peptide/lysozyme receptor is briefly outlined. PMID- 11401197 TI - The ontogenetic development of auditory sensitivity, vocalization and acoustic communication in the labyrinth fish Trichopsis vittata. AB - The anabantoid fish Trichopsis vittata starts vocalizing as 8-week-old juveniles. In order to determine whether juveniles are able to detect conspecific sounds, hearing sensitivities were measured in six size groups utilizing the auditory brainstem response-recording technique. Results were compared to sound pressure levels and spectra of sounds recorded during fighting. Auditory evoked potentials were present in all size groups and complete audiograms were obtained starting with 0.18 to 0.30 g juveniles. Auditory sensitivity during development primarily increased between 0.8 kHz and 3.0 kHz. The most sensitive frequency within this range shifted from 2.5 kHz to 1.5 kHz, whereas thresholds decreased by 14 dB. Sound production, on the other hand, started at 0.1 g and sound power spectra at dominant frequencies increased by 43 dB, while dominant frequencies shifted from 3 kHz to 1.5 kHz. Comparisons between audiograms and sound power spectra in similar-sized juveniles revealed no clear match between most sensitive frequencies and dominant frequencies of sounds. This also revealed that juveniles cannot detect conspecific sounds below the 0.31 to 0.65 g size class. These results indicate that auditory sensitivity develops prior to the ability to vocalize and that vocalization occurs prior to the ability to communicate acoustically. PMID- 11401198 TI - Auditory frequency discrimination in the squirrel monkey. AB - Four squirrel monkeys (Saimiri sciureus) were tested for their frequency discrimination capacity using an eyeblink classical conditioning procedure, with air puff against the eye as unconditioned stimulus and 600-ms pure tones as conditioned stimuli. Absolute frequency difference thresholds showed a minimum (20-41 Hz, mean 30 Hz) at 4,000-8,000 Hz and increased towards higher as well as lower frequencies (70-90 Hz, mean 80 Hz at 300 Hz; 44-120 Hz, mean 82 Hz at 16,000 Hz). Relative frequency difference thresholds increased from higher to lower frequencies, with values as low as 0.3-0.8% (mean 0.5%) at 16,000 Hz and as large as 24-30% (mean 27%) at 300 Hz. The squirrel monkey's frequency discrimination function thus shows a severe deviation from Weber's law. The frequency difference thresholds are comparable to human's in the 4,000-8,000 Hz range, but are 65-80 times higher in the 500- to 300-Hz range. Individuals with high auditory thresholds do not necessarily also have high frequency difference thresholds. PMID- 11401199 TI - Temperature compensation of postsynaptic currents from the extraocular muscle of temperate teleost fishes. AB - Miniature end plate currents were recorded from white inferior oblique extraocular muscle fibres of one temperate marine teleost (Aldrichetta forsteri, Family Mugilidae) and two temperate freshwater teleosts (Galaxias fasciatus, Family Galaxiidae and Oncorhynchus mykiss, Family Salmonidae). Miniature end plate currents were digitised and averaged over a temperature range of 5-25 degrees C. For each species, decay of miniature end plate currents was exponential and exhibited a strong temperature dependence. Lower temperatures resulted in prolonged decay phases, which decreased exponentially as a function of absolute temperature. Although values of the exponential time constant tau (tau) obtained for each species at 5 degrees C, 15 degrees C and 25 degrees C were significantly different, at any given temperature, there were no significant differences between tau values for the three species, despite differences in phylogeny (different families) and habitat (marine versus freshwater). At their normal temperature of 15 degrees C, mean values of tau for the three species ranged from 840 micros to 940 micros, and apparent activation energies ranged from -41 kJ mol(-1) deg(-1) to 50 kJ mol(-1) deg(-1). These observations confirm earlier reports that teleost miniature end plate currents are consistently shorter than those of other vertebrates. PMID- 11401200 TI - Electrophysiological responses of taste cells to nutrient mixtures in the polyphagous caterpillar of Grammia geneura. AB - In a normally feeding insect, the taste receptors are exposed to complex mixtures of chemicals, not single compounds. We investigate the responses of neurons in the galeal sensilla of the caterpillar of Grammia geneura to mixtures of nutrient compounds at concentrations occurring in plants. Compounds that stimulated the same neuron were generally additive in their effects in binary mixtures. Amino acids that did not stimulate usually had no effect in mixtures with a stimulating compound, but glutamic acid reduced the response to serine in the medial sensillum. Nutrient compounds that stimulated different cells in a sensillum acted independently of each other. Complex mixtures of amino acids resembling samples of free amino acids from three host plants were less stimulating than expected from their molar concentrations. In host plant selection, the response from the medial sensillum is probably dominated by sucrose; unless sucrose levels are low, amino acids will contribute little to sensory input because they stimulate the same cell as sucrose. In the lateral sensillum, amino acids act independently of sugars. The limited contact chemosensory array of caterpillars seems inadequate to allow them to make fine distinctions between plants on the basis of their free amino acids. PMID- 11401201 TI - Detection of coloured patterns by honeybees through chromatic and achromatic cues. AB - We asked whether the detection range of two-coloured centre-surround patterns differs from that of single-coloured targets. Honeybees Apis mellifera were trained to distinguish between the presence and absence of a single-coloured disc or a coloured pattern at different visual angles. The patterns presented colours which were either different in chromatic and L-receptor contrasts to the background, equal in chromatic but different in L-receptor contrasts, or vice versa. Patterns with colours presenting only chromatic contrast were also tested. Patterns with higher L-receptor contrast in its outer than in its inner element were better detected than patterns with a reversed L-contrast distribution. However, both were detected worse than single-coloured discs of the respective colours. When the L-receptor contrast was the same for both elements, the detection range of the two-coloured and single-coloured targets was the same. Patterns whose colours lacked L-receptor contrast were detected just as single coloured targets of the same colours. These results demonstrate that both chromatic and L-receptor contrasts mediate the detection of coloured patterns and that particular distributions of L-receptor contrast within a target are better detected than others. This finding is consistent with the intervention of neurons with centre-surround receptive fields in the detection of coloured patterns. PMID- 11401202 TI - Sound-localization experiments with barn owls in virtual space: influence of broadband interaural level different on head-turning behavior. AB - Interaural level differences play an important role for elevational sound localization in barn owls. The changes of this cue with sound location are complex and frequency dependent. We exploited the opportunities offered by the virtual space technique to investigate the behavioral relevance of the overall interaural level difference by fixing this parameter in virtual stimuli to a constant value or introducing additional broadband level differences to normal virtual stimuli. Frequency-specific monaural cues in the stimuli were not manipulated. We observed an influence of the broadband interaural level differences on elevational, but not on azimuthal sound localization. Since results obtained with our manipulations explained only part of the variance in elevational turning angle, we conclude that frequency-specific cues are also important. The behavioral consequences of changes of the overall interaural level difference in a virtual sound depended on the combined interaural time difference contained in the stimulus, indicating an indirect influence of temporal cues on elevational sound localization as well. Thus, elevational sound localization is influenced by a combination of many spatial cues including frequency-dependent and temporal features. PMID- 11401203 TI - Two thermosensors in Drosophila have different behavioral functions. AB - Insects inhabit extreme temperature environments and have evolved mechanisms to survive there. Small insects are especially susceptible to rapid changes in body temperature. Therefore, the rapid detection of environment and body temperature is important for their survival. Little, however, is known about the thermosensors that detect those temperatures. Using rapid thermosensitivity assays with temperature step gradients and a spatial learning paradigm (the heat box) in which elevated temperature serves as the negative reinforcer, two thermosensors were identified and their behavioral functions assessed. A low temperature thermosensor is located on the antenna, detects relatively low temperatures, and can detect spatial temperature gradients directly. Thus, the antennae can be used by Drosophila to quickly orient with respect to temperature cues. A high-temperature thermosensor of unknown location appears to have a roughly similar sensitivity to temperature differences as the low-temperature thermosensor (< or = 3 degrees C) and is both necessary and sufficient for memory formation in the heat-box spatial learning paradigm. Therefore, the high temperature thermosensor is important for remembering spatial positions in which dangerously high temperatures were encountered. PMID- 11401204 TI - Orientation and movement patterns of the wood mouse (Apodemus sylvaticus) in its home range are not altered by olfactory or visual deprivation. AB - Movements of wood mice (Apodemus sylvaticus) were recorded in the field in natural (non-deprived) and in vision- or olfaction-deprived conditions. Both visually and olfactorily deprived animals were able to efficiently orient themselves within their home ranges. There were no, or only nearly, significant differences between the movement characteristics of non-deprived and deprived animals. Moreover, movements of deprived animals were significantly more concentrated than expected in familiar places, i.e. places previously visited in non-deprived conditions (with respect to the null hypothesis that a deprived animal cannot recognize if it is moving through a familiar place). These results suggest that visually or olfactorily deprived wood mice recognize if the current place is familiar or not, and can move and orient themselves within their home ranges mostly as non-deprived wood mice do. PMID- 11401205 TI - Acute myeloid leukemia following chronic low-dose cyclophosphamide for metastatic breast cancer. AB - Unconventional treatments are commonly considered by the scientific community to have unproven efficacy but at least no toxicity. Here we report on the case of a breast cancer patient with lung and liver metastases who developed acute myeloid leukemia after treatment with Di Bella multitherapy, leading rapidly to death due to cerebral hemorrhage. Although an increased susceptibility to malignancy could not be excluded, we considered the possible etiologic role of the treatment received. The drug most likely to be associated with the development of leukemia was the cyclophosphamide contained in the Di Bella multitherapy regimen at a dose of 50 mg daily. The clinical features of this leukemia were therefore compared with those expected for secondary leukemia related to alkylating agents. A preceding myelodysplastic phase and the development of the leukemia after the intake of a cumulative cyclophosphamide dose of 15 g were typical chacteristics of secondary leukemia, but the interval between the start of therapy and the onset of leukemia was only 10 months. We conclude that long-term low-dose cyclophosphamide may have leukemogenic potential and the latency period may be shorter than that commonly reported. PMID- 11401206 TI - Recurrence of infantile supratentorial ependymoma after 23-year remission following surgical removal and radiation therapy. AB - We report on a patient with ependymoma who had a recurrence after long-term remission. The patient developed frontoparietal ependymoma at the age of one year and ten months. The tumor was radically removed and postoperative radiation therapy was performed. A calcified area adjacent to the area of surgical removal remained unchanged until the patient was 18 years old. The patient was healthy except for mild hemiparesis until an MRI scan performed when he was 25 years old showed regrowth of the tumor. The patient underwent surgery with additional radiation therapy and was discharged. The 23-year interval until tumor recurrence in this case is far beyond the so-called risk period of "Collins' law". Immunohistochemical study with MIB-1 and anti-p53 antibody showed a high proliferative potential of the primary and recurrent tumors and possible p53 mutation in the primary tumor. This is the first report to describe the detailed clinical course and histological features of a recurrent infantile ependymoma that progressed after Collins' risk period. It seems that follow-up of ependymoma patients after initial treatment should be performed regularly for a longer period in cases showing radiological evidence of a residual lesion. PMID- 11401207 TI - Peripheral primitive neuroectodermal tumor (PPNET) of pelvic origin: report of a case arising from an unusual location. AB - A peripheral primitive neuroectodermal tumor arising from the abdominopelvic cavity is reported in a 24-year-old young male without any previous remarkable pathology. He was admitted to our hospital with complaints of urinary symptoms (acute urinary retention) and mild intestinal occlusion that had been present for three months. Physical examination and CT scan revealed a pelvic mass occupying the entire pelvic cavity. The diagnostic workup included a CT-guided biopsy which defined the tumor as a sarcomatous type. Radical surgery was performed including tumor resection, pelvic exenteration (bladder and prostate gland) and urinary and fecal diversion. Adjuvant chemotherapy (VAIA) was delivered once the histology was confirmed. We reviewed the available literature focusing on the varied nomenclature of this tumor (peripheral neuroepithelioma, Askin's tumor, Ewing's extraosseous tumor, peripheral adult neuroblastoma, peripheral primitive extracranial neuroectodermal tumor (PPNET), the clinical features, the role of diagnostic imaging techniques, pathologic assessment and controversial therapeutic management. In addition, the prognosis and survival of this rare condition were analyzed. PMID- 11401208 TI - Correspondence re: Cicchetti S, Jemec B, Gault DT: two case reports of vinorelbine extravasation: management and review of the literature. Tumori, 86: 289-292, 2000. PMID- 11401209 TI - Peritoneal carcinomatosis from an unknown primary site. Management of 15 patients. AB - AIMS AND BACKGROUND: Peritoneal carcinomatosis from an unknown primary site is a rare and ill-defined entity. This work attempts to identify clinical and pathological features of patients with this disease and report the results of an aggressive combined treatment modality. METHODS: Retrospective analysis was performed of medical records of 15 patients with peritoneal carcinomatosis with no primary site identified at a single institution between 1989 and 2000. A primary gastrointestinal cancer was ruled out after a thorough endoscopic and radiologic work-up and complete exploratory surgery. RESULTS: Four women and 11 men were identified; the average age was 49 years. All patients had cytoreductive surgery with peritonectomies; 4 patients underwent a second-look operation. Perioperative intraperitoneal chemotherapy was given to 10 of the 15 patients, and 9 patients received post-cytoreduction chemotherapy given intraperitoneally (1), systemically (7) or both intraperitoneally and systemically (1). Overall median survival from diagnosis was 19.0 months; 1 patient is alive with disease at 21 months; and 3 patients are disease-free at 17, 38, and 60 months from diagnosis. Significant positive predictive factors for survival were a small volume of ascites (P = 0.02), a large number of peritonectomies performed (P = 0.001), second-look cytoreduction (P = 0.003), perioperative intraperitoneal chemotherapy (P = 0.008) and postoperative chemotherapy (P= 0.01), either intraperitoneal or systemic. CONCLUSIONS: Peritoneal carcinomatosis from an unknown primary site is a rare subset of primary peritoneal malignancy. Aggressive treatment may provide prolonged palliation with occasional long-term survival. PMID- 11401210 TI - The clinicopathologic characteristics of colorectal cancer in patients under 50 years of age: experience of an oncology center. AB - BACKGROUND/AIM: Colorectal cancer is seen mostly among patients older than 50 years of age. An aggressive behavior is a frequently cited as characteristic of colorectal cancer in young patients. The purpose of the present study was to reveal the clinicopathologic characteristics of colorectal cancer among patients under 50 years of age. METHODS: Two hundred and seventy-one patients with colorectal cancer admitted to our oncology center were evaluated, and clinicopathologic findings of the young and old patients were compared. Patient gender, site distribution, tumor stage classification, lymph node involvement, metastatic site, histologic classification, histologic differentiation, family history of malignant tumors, presenting symptoms and survival rates were compared. RESULTS: One hundred patients were 50 years of age or under. Clinical, histopathologic characteristics and overall survival of the two groups did not differ. A higher rate of familial cancer syndromes was detected among young patients. CONCLUSIONS: The presentation and outcome of the disease in young patients do not differ from those of older patients. A significant family history of colorectal cancer in the young patients showed the need for screening whereas the outcome of metastatic disease was poor. In order to anticipate long survival, early detection and aggressive treatment is necessary. PMID- 11401211 TI - Quality of life in patients with early stage breast carcinoma treated with conservation surgery and radiotherapy. An Italian monoinstitutional study. AB - AIMS: To evaluate the quality of life (QOL) in patients with early stage invasive carcinoma of the breast treated with conservative surgery and postoperative irradiation. METHODS: A mailed survey to examine QOL was conducted in 227 subjects with breast cancer treated in 1990 and 1994 with conservation surgery plus definitive irradiation. The self-compiled questionnaire was developed based on a series of 38 items assessing six core areas of QOL. Furthermore, the patients were requested to evaluate the degree of information provided by the medical staff concerning the disease, the treatment and related side effects and to evaluate the effects of the treatment on their social, overall QOL, and health status. RESULTS: The questionnaire was completed by 156 patients (68.7%) who had a median age of 56 years (range, 28-75 years) at the time of treatment and 59 years (range, 31-82 years) at the time of the study. The physical condition was reported to be good. Data relating to sexual life were provided by more than 90% of the patients. Some limitations in sexuality, some interference with sexual desire, and some modifications during intercourse were reported by 11, 11, and 10 patients, respectively. The subjective evaluations of the cosmetic results of the therapies were judged good-excellent by 56% of the patients, 12 (8%) only had a negative perception of their body image. Twenty-five percent of the sample declared that they felt tense, 19% nervous, 18% lonely, 27% anxious and 16% depressed. Only six patients (4%) declared that the treatment had had a bad on their social life, and 18 (11%) thought that their health status has been affected by the treatment. A worsening of QOL due to the disease or the treatment was reported by 8% of the responders. The amount of information received concerning the treatment and its side effects was considered sufficient by most of the patients. CONCLUSIONS: The results of the study revealed a satisfactory health-related QOL in patients treated with breast conservation and postoperative irradiation. A preserved favorable body image and lack of a negative impact on sexuality was observed, even though about half of the patients reported a negative judgement on esthetic outcome. Some patients had persistent psychosocial concerns. No significant additional problems attributable to radiation therapy capable of affecting QOL outcomes were reported. PMID- 11401212 TI - Irradiation fields and doses in glioblastoma multiforme: are current standards adequate? AB - AIMS AND BACKGROUND: The optimum conventional radiotherapy in glioblastoma multiforme patients has not been clearly defined by prospective trials. To better characterize a standard radiotherapy in glioblastoma multiforme, the impact on survival of different fields and doses was analyzed in a retrospective single center series. METHODS: One hundred and forty-seven patients with glioblastoma multiforme, submitted to biopsy only (n = 15), subtotal (n = 48) or total resection (n = 82) and who completed the planned postsurgical radiotherapy, were considered. The median age was 57 years, the male/female ratio 1.5/1, and the performance status > or =70 in 76%. Whole brain irradiation, followed by a boost to partial brain, was used in 75 cases with a whole brain dose of 44-50 Gy (median, 46) and a partial brain dose of 56-70 Gy (median, 60 Gy). Partial brain irradiation alone was used in 72 patients with a dose of 56-70 Gy (median, 61 Gy). Ninety-eight patients received 56-60 Gy (median, 59 Gy) to partial brain whereas 49 patients received 61-70 Gy (median, 63 Gy). RESULTS: There was an almost significantly longer survival in patients irradiated to the partial brain alone with respect to those also receiving whole brain radiotherapy (P = 0.056). Doses >60 Gy significantly prolonged survival (P = 0.006). Multivariate analysis confirmed that the impact on survival of radiation dose was independent of age, performance status, extent of surgery, field of irradiation and the use of chemotherapy. The extent of irradiation field was not independently related to improved survival. CONCLUSIONS: Our retrospective findings suggest that we reflect on the adequacy of the current standard irradiation parameters. Well designed prospective trials are necessary to standardize the radiotherapy control group in patients with glioblastoma multiforme to be compared in phase III trials with innovative therapeutic approaches. PMID- 11401213 TI - The role of a conventional simulator in multileaf-plan simulation: a proposal. AB - Modern computer networks provide satisfying levels of data recording and verification between the treatment planning system (TPS) and the accelerators, while the main weakness of the preparation chain remains the simulation. When a conventional simulator is employed, it may adversely affect the three-dimensional treatment planning system (3DPS) process because of the difficulty to document the leaf positions on the simulator location films and on the patient's skin. With a conventional simulator, hard copies of the DRRs of each field and CT scans at isocenter level are needed. In an attempt to transfer more information displayed from a BEV perspective from the 3DPS to simulator radiographs, this study aimed to reduce the quality loss by using a 2D conventional simulator in a 3DPS process. We realized an acetate photocopy of TPS data for each field, from a BEV perspective, containing: DRR, wire frames of the PTV, organs at risk and MLC aperture. The photocopies, with an appropriate magnification factor to obtain a correct projective value (ratio 1:1) at isocenter level, are carefully placed on the radiographic images on the same hard copy which allows us to better understand possible setup errors and obliges us to correct these. The method provides reliable documentation, facilitates treatment verification, and fulfils the criteria for MLC simulation. It is accurate, simple, and very inexpensive. PMID- 11401214 TI - Reproducibility of histopathologic diagnosis and classification of non melanocytic skin cancer: a panel exercise in the framework of the multicenter southern European study HELIOS. AB - AIMS AND BACKGROUND: The reproducibility of histologic diagnoses, and in particular of the distinction between basal and squamous cell forms, has been evaluated in the framework of a multicenter case-control study (in Italy, France, Spain and Switzerland) aimed to assess the causes of non-melanocytic skin cancers. METHODS: A panel composed of 10 pathologists from the collaborating centers was appointed. A total of 1,774 slides of routine diagnoses were blindly reviewed by a second panelist; discordant diagnoses underwent a third examination. Controversial and interesting cases were discussed during general sessions. RESULTS: The results showed a high degree of concordance (99.5%), with a Cohen's kappa of 0.85 (95% CI, 0.77-0.94) in the assessment of malignancy of lesions. The concordance in the distinction between large morphologic groups, basal cell carcinoma and squamous cell carcinoma was also high (Cohen's kappa = 0.85; 95% CI, 0.82-0.89). The assessment of histologic subtypes, degree of invasion and differentiation showed a lower degree of concordance, presumably as a consequence of a weaker consistency in the relevant classifications. CONCLUSIONS: The reproducibility study has therefore validated the epidemiologic study and in particular allowed to correct some misclassifications that could have lessened the analysis of the case-control study. In general, because of its characteristics (number of pathologists and variety of their origins, the large number of cases examined, blind examination of histologic slides), the conclusions of the study may show a certain degree of generalization, at least with regard to the countries represented. Routine histologic diagnoses of cutaneous carcinoma showed a high degree of reliability with reference to the assessment of malignancy and the distinction between basal and squamous cell carcinoma morphotypes. PMID- 11401215 TI - Thrombocytopenia and its correction by platelet concentrates. PMID- 11401216 TI - Italian guidelines for the appropriate use of plasma. PMID- 11401217 TI - The immunomodulatory effects of blood transfusion. PMID- 11401218 TI - Noninfective complications of blood transfusions. PMID- 11401219 TI - Management of venous thrombosis in cancer. Recent developments in diagnosis, prevention, and treatment. PMID- 11401220 TI - Hepatic venoocclusive disease: a major complication of hematopoietic stem cell transplantation in cancer patients. PMID- 11401221 TI - Indications for autotransfusion in surgical oncology. PMID- 11401222 TI - Leukocyte-depleted blood components. PMID- 11401223 TI - Peripheral blood stem cell collection, preparation and selection. PMID- 11401224 TI - Anemia of cancer: an overview. PMID- 11401225 TI - High throughput HLA sequence-based typing (SBT) utilizing the ABI Prism 3700 DNA Analyzer. AB - AIMS AND BACKGROUND: The genetic complexity of the human major histocompatibility complex (MHC) has required the development of various molecular typing methods. The purpose of this paper is to compare the results of two of these molecular methods: sequenced based typing (SBT) and polymerase chain reaction (PCR) using sequence specific primers (PCR-SSP). METHODS: The SBT method described utilizes an ABI Prism 3700 DNA Analyzer, which has been designed fro high throughput production of sequence data through highly automated operation with significant walk-away time. The ABI Prism 3700 DNA Analyzer is a 96-capillary electrophoresis instrument with the capability of running four 96-well plates black to back in a sixteen-hour period. Potentially, data from this machine can produce Class I sequences for A or B loci for 64 samples in this time frame. The SBT method encompassed exons 2, 3, and 4 with forward and reverse sequence orientation reactions using the PE Biosystems HLA-A and HLA-B Sequenced Based Typing Kits (PE Applied Biopsystems/Perkin-Elmer, Foster City, CA, USA). Most SBT methods previously employed only gather data from exons 2 and 3 which distinguishes most of the polymorphism necessary to identify the majority of alleles in the HLA region. However, in an effort to discern numerous null alleles in the HLA region, exon 4 data is also included. The PCR-SSP method utilized consists of one 96 well tray, with 95 primer mixes and one negative control, per sample designed to produce an intermediate/high resolution HLA-A, B typing. RESULTS: Data from one 96-well capillary run on the ABI Prism 3700 DNA Analyzer, which consists of results from 16 samples for HLA-A or HLA-B loci, was compared to data derived from sixteen HLA-A and HLA-B PCR-SSP typings. 75% of loci tested achieved a higher resolution HLA typing by the SBT method. DISCUSSION: The ability to provide allele level HLA typing results can have significant functional implications for the bone marrow transplant community and numerous vaccine studies. PMID- 11401226 TI - The health and safety of the donors. PMID- 11401227 TI - Screening and diagnosis of blood-borne infections in Italy. PMID- 11401228 TI - The European dimension of voluntary blood donation. PMID- 11401229 TI - Transfusion transmitted infections. PMID- 11401230 TI - The right to privacy and access to information about one's genetic origins. AB - The right to privacy contains different dimensions. Best known are the possibility to restrict the flow of personal information to others (limited access by others) and the right not to know certain information gathered about oneself. Since the privacy right is meant to protect the identity, self-image and personal life of the person, he should also have the right to know certain facts about himself. Information about genetic origins is potentially important for the child conceived by means of donor insemination because it tells him something about himself and about the relationships he has with others. However, the positive privacy right of the donor offspring may be overridden by the negative privacy right of the parents in which case they may keep the method of conception a secret and by the negative privacy of the donor which gives him the right to remain anonymous. PMID- 11401231 TI - Should Canadian physicians have a mandatory duty to report woman abuse? AB - As in other countries, woman abuse is a leading public health issue in Canada. In recognition of its seriousness and the need to interrupt the cycle of violence, several jurisdictions outside of Canada have enacted mandatory reporting laws for woman abuse that require health care workers to report cases. This article reviews the major debates surrounding mandatory reporting of woman abuse with an emphasis on where knowledge is incomplete but would be of benefit when weighing the pros and cons of such laws. We conclude with comments and recommendations with respect to Canada and its health legislation. PMID- 11401232 TI - How can we avoid some psychotic patients becoming violent offenders? AB - Every year a number of psychotic patients commit violent crimes and are recommended for forensic psychiatric treatment. Many of these patients already show risk symptoms before their fatal offense. Despite these risk signs the physicians and psychiatrists, who have the task of intervening in crisis situations of psychotic patients, are frequently waiting too long before compulsory admission of potentially dangerous psychotic patients to a general psychiatric hospital. A physician or psychiatrist, who admits a "harmless" patient against his or her will may risk prosecution. Therefore, many involved professionals are very careful with regard to a forced admission of problematic psychotic patients to a general psychiatric hospital. It is timely, that some risk signs, which are evidently linked to a significantly, increased risk of violent behavior in psychotic patients, become recognized officially by law as a good reason for compulsory admission to a general psychiatric hospital. PMID- 11401233 TI - Ethical acceptability, health policy and foods biotechnology based foods: is there a third way between the precaution principle and an overly enthusiastic dissemination of GMO? AB - The demand for consumer safety with regard to the food-processing industry is becoming, legitimately, more and more urgent. If ingested drugs can carry deleterious effects that exceed the beneficial effect that the research was initially undertaken for, then the same can only be the case for foods that stem from the same new biotechnologies, zero risk being non existent. There are two conflicting viewpoints about the possible risks linked to genetically modified organisms: a posteriori protection (based on vigilance once the product is on the market) and an a priori protection (at present usually supported by the precaution principle). We suggest a third way, which ensures consumer safety, but doesn't hinder scientific progress. Just as there are regulations for the protection of human subjects in biomedical research and regulations for the use of drugs after they are marketed, so should such regulations be introduced in the domains of food production that use biotechnologies. We therefore suggest that the scientific community and the food-processing industry develop evaluation protocols for new foods like the ones that exist for drugs. We thus offer thirteen regulations, based on the Helsinki declaration, in order to establish these protocols. These proposals, applied to food-processing research, would enable the industry to return confidence to consumers and thus avoid the random blocking of scientific progress, which is a source of health for the greater population. PMID- 11401234 TI - Patients' experiences and views of clinical trials. AB - This article explores whether the experience of participating in a clinical trial was similar to what patient volunteers had expected, prior to agreeing to take part. It also considers patients' views on four aspects of participation that may help to determine whether the decision to become a research subject was truly an autonomous one, namely: whether patients feel that (i) doctors 'know best' when it comes to taking part in a trial; (ii) refusing to take part would upset the doctor; (iii) there is little risk associated with participation, and (iv) people have a moral duty to be involved in such trials. FINDINGS: More than a third of patients had no particular expectations of what trial participation would be like. Over half did not believe that doctors knew best about whether or not they should participate, and the vast majority rejected the proposition that a refusal to participate would have upset the doctor. Half of the patients believed that there was little risk in testing new drugs, and most felt that people do not have a moral duty to become research subjects. PMID- 11401235 TI - Handicap assessment: setting the grounds for an effective intervention in the community. AB - Many instruments have been invented specially since the 70's to study and measure the consequences of illness and trauma; many of them simply evaluate the severity of lesions and the prediction of mortality, others evaluate morbidity. Nevertheless, none of these instruments evaluate the handicap globally. The classification of the World Health Organization, despite the fact that it contemplates the handicap in all its aspects, does not distinguish its different levels rigorously and it is not a valid instrument to quantify handicaps. The authors comment on their research and proposals for identification and quantification of the handicap. Three levels, related to "Body", "Capacities" and "Life situations", have been described and through them a useful inventory was created (published in 1998). This inventory makes it possible to establish a true medical-social and global evaluation of handicaps, providing a uniform record of the consequences of diseases, trauma or physiological features of the individual and a useful instrument for public health intervention and the promotion of human well-being. PMID- 11401236 TI - A survey of patients' rights representatives in Israeli Hospitals: 1999-2000. AB - Israel's legislature enacted a Patients' Rights Act in 1996. The Act directed hospitals to designate a representative responsible for supervising protection of patients' rights under the new law, investigating complaints, and educating medical staff regarding the provisions of the Act. This survey was conducted in order to evaluate implementation of that section of the law pertaining to hospital representatives of patients' rights. The authors conducted interviews with responsible individuals at government hospitals in Israel, inquiring about their duties, training, complaint procedures, and practices. The authors found that while the representatives shared some basic knowledge of their legal duty, methods of implementation differed widely. It is recommended that a comprehensive program of training be devised for hospital patients' rights representatives, in order to realize the potential of the Act. PMID- 11401237 TI - Genetics, data protection and non-discrimination: some reflections from an international human rights law perspective. AB - As a result of progress in medical research genetic traits can increasingly be used as predictive factors and selection criteria. This calls into question the appropriateness, validity and legitimacy of genetic traits as (co-)determinants in decision-making processes. This paper confines itself to examining the implications of genetics on the right to privacy--notably data protection--and the right not to be discriminated against. PMID- 11401238 TI - Health care workers with AIDS: the patient's right to know. AB - This paper discusses the relationship between a hospital patient and his healthcare worker who is HIV positive or has active AIDS. What are the actual and potential risks involved? How much is a patient entitled to know about the HIV status of his doctor, dentist or nurse? How much does a carer have to reveal in this regard to a patient? Case law is adduced and discussed in the light of current approaches to this problem. PMID- 11401239 TI - Physicians' attitudes towards patients' rights legislation. AB - Patients' rights laws, bills and charters aim at delineating the patient physician relationship in regard to consent to medical treatment, confidentiality and related issues. The need to shape such an intimate relationship by way of legislation seems anomalous to some, but imperative to others. We present for the first time an insight into Israeli physicians' attitudes towards Israel's patients' rights laws, in a changing medical and socio-legal environment. The research results suggest that physicians are reluctant to participate in the implementation of such laws, demonstrated by the level of their misunderstanding of the law's norms and regulations, and subjective attitudes and perceptions. In order to ensure the medical community's participation in augmenting patients' rights, efforts should focus on improved legal and ethical education, enhanced cooperation of professional associations and joint action with legislators to assure a productive composition of these important acts. PMID- 11401240 TI - Acute care crisis simulation for jury education. AB - A major impediment to appropriate adjudication in medical malpractice cases is the large difference in knowledge and background of the expert witness compared to judges, other litigants and juries. Such disparities make it plausible for an expert witness to support an issue, even in defiance of common sense. The most basic understanding of the atmosphere and practices surrounding critical events and the routine procedures, techniques and equipment which are employed in critical situations is absent. (McAbee GN: Improper expert medical testimony: existing and proposed mechanisms of oversight. Journal of Legal Medicine 19: 257 272, 1998). PMID- 11401241 TI - The fitness of the mental patient to be a witness. AB - The determination of whether a patient has criminal responsibility or is able to stand trial is routine psychiatric work. Cases in which we- psychiatrists- are asked to express our opinion on whether a patient can testify, however, are quite rare. We shall attempt to clarify some of the issues relating to the testimony of mentally ill patients through a case presentation. In this case, the Court agreed to consider our patient's testimony, only after receiving our expert opinion. We find that the Court's reversal of its original decision and its willingness to consider the testimony of a mental patient in the same trial in which he was initially found unable to stand trial, marks an important precedent. The question of the credibility of the mental patient as a witness has significance beyond this case concerning mental patients' rights in regard to the judicial system. We believe there is a need for further discussion between psychiatrists andjurists regarding the fitness of the mental patient to testify as a witness. PMID- 11401242 TI - The insanity defense: examination of the extent of congruence between psychiatric recommendation and adjudication. AB - The insanity plea is a known defense often utilized in courts of law. In such cases the accused may be referred for an outpatient psychiatric examination or for hospital observation. In this study, we examined the extent of the accord between the medical recommendations of the forensic unit of the Yehuda Abarbanel Mental Health Center and judicial decisions. It was found that in 99.4% of the cases, the court accepted the psychiatric recommendation. In only 2 cases the recommendations were not accepted. We discuss this issue and recommend improvements and strengthening of the relationship between the psychiatric and court systems. PMID- 11401243 TI - Criteria document for evaluating the work-relatedness of upper-extremity musculoskeletal disorders. PMID- 11401244 TI - Delay of natural bone loss by higher intakes of specific minerals and vitamins. AB - For early prevention or inhibition of postmenopausal and age-related bone loss, nutritional interventions might be a first choice. For some vitamins and minerals an important role in bone metabolism is known or suggested. Calcium and vitamin D support bone mineral density and are basic components in most preventive strategies. Magnesium is involved in a number of activities supporting bone strength, preservation, and remodeling. Fluorine and strontium have bone-forming effects. However, high amounts of both elements may reduce bone strength. Boron is especially effective in case of vitamin D, magnesium, and potassium deficiency. Vitamin K is essential for the activation of osteocalcin. Vitamin C is an important stimulus for osteoblast-derived proteins. Increasing the recommended amounts (US RDA 1989), adequate intakes (US DRI 1997), or assumed normal intakes of mentioned food components may lead to a considerable reduction or even prevention of bone loss, especially in late postmenopausal women and the elderly. PMID- 11401245 TI - Chemical composition and potential health effects of prunes: a functional food? AB - Prunes are dried plums, fruits of Prunus domestica L., cultivated and propagated since ancient times. Most dried prunes are produced from cultivar d'Agen, especially in California and France, where the cultivar originated. After harvest, prune-making plums are dehydrated in hot air at 85 to 90 degrees C for 18 h, then further processed into prune juice, puree, or other prune products. This extensive literature review summarizes the current knowledge of chemical composition of prunes and their biological effects on human health. Because of their sweet flavor and well-known mild laxative effect, prunes are considered to be an epitome of functional foods, but the understanding of their mode of action is still unclear. Dried prunes contain approximately 6.1 g of dietary fiber per 100 g, while prune juice is devoid of fiber due to filtration before bottling. The laxative action of both prune and prune juice could be explained by their high sorbitol content (14.7 and 6.1 g/100 g, respectively). Prunes are good source of energy in the form of simple sugars, but do not mediate a rapid rise in blood sugar concentration, possibly because of high fiber, fructose, and sorbitol content. Prunes contain large amounts of phenolic compounds (184 mg/100 g), mainly as neochlorogenic and chlorogenic acids, which may aid in the laxative action and delay glucose absorption. Phenolic compounds in prunes had been found to inhibit human LDL oxidation in vitro, and thus might serve as preventive agents against chronic diseases, such as heart disease and cancer. Additionally, high potassium content of prunes (745 mg/100 g) might be beneficial for cardiovascular health. Dried prunes are an important source of boron, which is postulated to play a role in prevention of osteoporosis. A serving of prunes (100 g) fulfills the daily requirement for boron (2 to 3 mg). More research is needed to assess the levels of carotenoids and other phytochemicals present in prunes to ensure correct labeling and accuracy of food composition tables in order to support dietary recommendations or health claims. PMID- 11401246 TI - Agricultural produces: synopsis of employed quality control methods for the authentication of foods and application of chemometrics for the classification of foods according to their variety or geographical origin. AB - A review of quality control methods and applications of multivariate statistical techniques on the authentication and classification of agricultural products is presented. The products reported within the frame of this article were vegetables, fruits, juices, jams, wines, cereals, bakery products, oils, tea, coffee, honey, sugar-syrups, salad dressings, and gums. The perspective of multivariate statistics as a promising tool to authenticate and classify these food products according to their geographical origin or variety was demonstrated. Several representative figures and informative synoptical tables for agricultural food products were provided both for the quality control methods employed and the multivariate analyses implemented. PMID- 11401247 TI - Phosphorus losses in furrow irrigation runoff. AB - Phosphorus (P) often limits the eutrophication of streams, rivers, and lakes receiving surface runoff. We evaluated the relationships among selected soil P availability indices and runoff P fractions where manure, whey, or commercial fertilizer applications had previously established a range of soil P availabilities on a Portneuf silt loam (coarse-silty, mixed, superactive, mesic Durinodic Xeric Haplocalcid) surface-irrigated with Snake River water. Water soluble P, Olsen P (inorganic and organic P), and iron-oxide impregnated paper extractable P (FeO-Ps) were determined on a 0.03-m soil sample taken from the bottom of each furrow before each irrigation in fall 1998 and spring 1999. Dissolved reactive phosphorus (DRP) in a 0.45-microm filtered runoff sample, and iron-oxide impregnated paper-extractable P (FeO-Pw), total P, and sediment in an unfiltered runoff sample were determined at selected intervals during a 4-h irrigation on 18.3-m field plots. The 1998 and 1999 data sets were combined because there were no significant differences. Flow-weighted average runoff DRP and FeO-Pw concentrations increased linearly as all three soil P test concentrations increased. The average runoff total P concentration was not related to any soil P test but was linearly related to sediment concentration. Stepwise regression selected the independent variables of sediment, soil lime concentration, and soil organic P extracted by the Olsen method as related to average runoff total P concentration. The average runoff total P concentration was 1.08 mg L(-1) at a soil Olsen P concentration of 10 mg kg(-1). Soil erosion control will be necessary to reduce P losses in surface irrigation runoff. PMID- 11401248 TI - Factors affecting alkalinity generation by successive alkalinity-producing systems: regression analysis. AB - Use of successive alkalinity-producing systems (SAPS) for treatment of acidic mine drainage (AMD) has grown in recent years. However, inconsistent performance has hampered widespread acceptance of this technology. This research was conducted to determine the influence of system design and influent AMD chemistry on net alkalinity generation by SAPS. Monthly observations were obtained from eight SAPS cells in southern West Virginia and southwestern Virginia. Analysis of these data revealed strong, positive correlations between net alkalinity generation and three variables: the natural log of limestone residence time, influent dissolved Fe concentration, and influent non-Mn acidity. A statistical model was constructed to describe SAPS performance. Subsequent analysis of data obtained from five systems in western Pennsylvania (calibration data set) was used to reevaluate the model form, and the statistical model was adjusted using the combined data sets. Limestone residence time exhibited a strong, positive logarithmic correlation with net alkalinity generation, indicating net alkalinity generation occurs most rapidly within the first few hours of AMD-limestone contact and additional residence time yields diminishing gains in treatment. Influent Fe and non-Mn acidity concentrations both show strong positive linear relationships with net alkalinity generation, reflecting the increased solubility of limestone under acidic conditions. These relationships were present in the original and the calibration data sets, separately, and in the statistical model derived from the combined data set. In the combined data set, these three factors accounted for 68% of the variability in SAPS systems performance. PMID- 11401249 TI - Relationships between biosolids treatment process and soil phosphorus availability. AB - Laws mandating phosphorus (P)-based nutrient management plans have been passed in several U.S. Mid-Atlantic states. Biosolids (sewage sludge) are frequently applied to agricultural land and in this study we evaluated how biosolids treatment processes and biosolids P tests were related to P behavior in biosolids amended soils. Eight biosolids generated by different treatment processes, with respect to digestion and iron (Fe), aluminum (Al), and lime addition, and a poultry litter (PL), were incubated with an Elkton silt loam (fine-silty, mixed, active, mesic Typic Endoaquult) and a Suffolk sandy loam (fine-loamy, siliceous, semiactive, thermic Typic Hapludult) for 51 d. The amended soils were analyzed at 1 and 51 d for water-soluble phosphorus (WSP), iron-oxide strip--extractable phosphorus (FeO-P), Mehlich-1 P and pH. The biosolids and PL were analyzed for P, Fe, and Al by USEPA 3050 acid-peroxide digestion and acid ammonium oxalate, Mehlich-1, and Mehlich-3 extractions. Biosolids and PL amendments increased extractable P in the Suffolk sandy loam to a greater extent than in the Elkton silt loam throughout the 51 d of the incubation. The trend of extractable WSP, FeO-P, and Mehlich-1 P generally followed the pattern: [soils amended with biosolids produced without the use of Fe or Al] > [PL and biosolids produced using Fe or Al and lime] > [biosolids produced using only Fe and Al salts]. Mehlich-3 P and the molar ratio of P to [Al + Fe] by either the USEPA 3050 digestion or oxalate extraction of the biosolids were good predictors of changes in soil-extractable P following biosolids but not PL amendment. Therefore, the testing of biosolids for P availability, rather than total P, is a more appropriate tool for predicting extractable P from the biosolids-amended soils used in this study. PMID- 11401250 TI - TurfPQ, a pesticide runoff model for turf. AB - Environmental assessments of golf courses and other turf systems must often rely on mathematical modeling. However, in the case of pesticide runoff, successful modeling applications are rare. Available models were developed for agricultural applications and have seen very limited testing for turf. TurfPQ is a pesticide runoff model developed exclusively for turf. The model is based on a curve number calculation for runoff volume and linear partitioning of pesticide into adsorbed and dissolved components during a precipitation or irrigation event. Calibration is optional, so the model can be applied, using default parameter values, to situations where runoff and chemical loss data are unavailable. TurfPQ was tested with default parameter values for 52 pesticide runoff events involving six pesticides measured in plot studies in four states. The model typically produced conservative overpredictions of pesticide runoff, particularly with strongly adsorbed pesticides. Mean predicted pesticide runoff was 2.9% [corrected] of application, compared with an observed mean of 2.1%. TurfPQ captured the dynamics of the pesticide runoff events well with R2 = 0.65 [corrected]. Sensitivity analyses indicated that prediction errors could be reduced by better estimates of adsorption parameters and runoff curve numbers. However, even with default parameters, TurfPQ predictions are at least as accurate as those produced by more complex models. PMID- 11401251 TI - Aging effects on cadmium transport in undisturbed contaminated sandy soil columns. AB - Limited information is available on the effects of contaminant aging (i.e., the contact time of Cd with the soil) on Cd transport in soils. We conducted displacement experiments in which indigenous Cd and freshly applied Cd were leached simultaneously from undisturbed samples of three Spodosol horizons. Sorption of Cd was described using Freundlich isotherms, whereas transport was described as a convection-dispersion process. Parameter optimization analysis using a mobile-immobile transport model applied to nonsorbing tracer displacement data showed that 16 to 22% of the water in the columns was immobile. The low dimensionless mass transfer coefficients in the mobile-immobile model were indicative of diffusion-limited transfer between mobile and immobile water, and hence physical nonequilibrium. A two-site kinetic sorption model could be fitted closely to breakthrough curves of the non-aged Cd for three soil horizons. No conclusive evidence was found that contaminant aging in soil affects cadmium transport. On the one hand, predictions of aged Cd leaching, using parameters estimated from displacement experiments with nonaged Cd, differed from those for the aged Cd in the E horizon. On the other hand, no meaningful differences in transport behavior between aged and non-aged Cd were found for the humus Bh and Bh/C horizons. The two-site kinetic rate coefficient alphac was found to depend on water flux, further indicating that mass transfer between sorption sites and the liquid is limited by diffusion rather than by kinetic sorption. PMID- 11401252 TI - Denitrification losses from outdoor piglet production: spatial and temporal variability. AB - Animal welfare considerations have stimulated the development of outdoor piglet (Sus scrofa) production systems, but the high levels of nutrients excreted suggest that nutrient losses from this system may be high. This study first described the spatial distribution of denitrification activity in a 5- x 5-m grid within and outside a paddock immediately after the sows (32 sows ha(-1) for 6 mo) were removed in October 1997, and again the following March. Denitrification rates averaged 0.01 kg N ha(-1) d(-1) outside, and 0.5 kg N ha(-1) d(-1) inside the paddock in October, while the corresponding figures in March were 0.01 and 0.1 kg N ha(-1) d(-1). The highest denitrification rates were observed around the feeder, and this was also the case for concentrations of dissolved organic C and inorganic N in the soil. A statistical model that included both soil parameters and distance to feeder and huts gave the best description of the variability, but there was no significant autocorrelation between sampling points. In a second phase, seasonal variation of denitrification activity within a paddock (12 sows ha(-1) yr(-1)) was quantified; 10 soil cores were sampled along a transect 11 times between March 1998 and February 1999. There was a significant positive effect of dissolved organic carbon (DOC) on denitrification at <25% gravimetric soil moisture (i.e., to November in this study). Both climate and management (position of huts and feeder) appeared to influence denitrification, which was estimated to be 69 kg N ha(-1) yr(-1), or 11% of the N surplus of this production system. PMID- 11401253 TI - Biosolids effects on phosphorus retention and release in some sandy Florida soils. AB - The soil solid phase components most responsible for P sorption in Florida soils are Fe and Al oxides. Thus, we hypothesized that land application of biosolids would significantly increase a soil's P retention by increasing its content of P sorbing solids, especially when biosolids with high Fe and Al concentrations are applied to soils that sorb P poorly. Biosolids effects were quantified by a series of single-point isotherms on soils from two field studies sampled for up to 4 yr after initial biosolids application. Biosolids additions had little effect on P retention in a soil with abundant oxalate-extractable Fe and Al and a correspondingly large native P-sorbing capacity. However, biosolids significantly increased P retention in a soil with low oxalate-extractable Fe and Al content and low native P-sorbing capacity. Biosolids effects on P retention lasted 1 to 3 yr after application, depending on biosolids source and rate of application, and generally mimicked persistence of increased extractable Fe and Al concentrations in the poorly P-sorbing soil. Disappearance of added Fe and Al (and, hence, P retention capacity) from the surface horizons over time was relatively rapid, perhaps due to abundant organic acid production associated with biosolids degradation. Phosphorus in biosolids containing (or tailored to contain) abundant Fe and/or Al can be expected to behave as a slowly available P source, and to be less subject to leaching losses than completely soluble P sources. PMID- 11401254 TI - Tree species for recovering nitrogen from dairy-farm effluent in New Zealand. AB - Land treatment of dairy-farm effluent is being widely adopted as an alternative to disposal into surface waters in New Zealand. This study investigated water balances and associated N leaching from short-rotation forest (SRF) species irrigated with dairy-farm effluent. Single trees were grown in lysimeters filled with Manawatu fine sandy loam (mixed mesic Dystric Eutrochrept). Dairy-farm effluent was applied during two irrigation periods at 21.5 mm wk(-1) with a total loading equivalent to 870 kg N ha(-1) occurring over 17 mo. Following tree harvest in April 1997, measurements continued until August 1997 to monitor tree reestablishment. Cumulative N leached did not differ between lysimeters in which evergreen Sydney blue gum (Eucalyptus saligna Sm.) and shining gum [Eucalyptus nitens (H. Deane & Maiden) Maiden] and deciduous kinu-yanagi (Salix kinuyanagi Kimura) were grown. Leachate N concentrations of all treatments were on average higher than the New Zealand drinking water standard of 11.3 mg N L(-1). The E. nitens and S. kinuyanagi treatments leached 33 and 35 kg N ha(-1) yr(-1) in 1996 following application of 236 kg N ha(-1) during the first irrigation season. Leaf area was strongly correlated to evapotranspiration, drainage volume, and nitrogen leached. The majority of leaching in the tree treatments occurred after harvest. Reducing the leaching in the regrowth phase may be achieved through timing harvest in the spring when growth rates are higher and leaching potential is lower. Based on N uptake rates observed in this study and average pond discharge, a plantation of 5.4 ha would be required for N recovery on a typical dairy farm in New Zealand. PMID- 11401255 TI - Effect of flue gas desulfurization residue on plant establishment and soil and leachate quality. AB - Effects on soil quality and crop establishment after incorporation of flue gas desulfurization by-product (FGD) into soil as an amendment was assessed in a mesocosm study. Mesocosm units received applications equivalent to 0, 2.5, 5.0, 7.5, and 10% FGD residue [0, 25, 50, 75, and 100 tons acre(-1)]. Germination, biomass production, and elemental composition of corn (Zea mays L. var. Dekalb DK 683), soybean [Glycine max (L.) Merr. var. Haskell Pupa 94], radish (Raphanus sativus L. var. Sparkler), and cotton (Gossypius hirsutus L. var. Deltapine 51) were determined. The quality of leachates and soil were also determined periodically. Flue gas desulfurization residue did not affect germination and all application rates stimulated aboveground biomass. Plants grown in FGD-amended soil contained significantly elevated tissue concentrations of As, B, Se, and Mo. The FGD residue elevated surface soil pH from 5.5 (Control) to 8.1 (at 10% FGD). Leachate pH was unaffected by FGD, but salinity rose sharply with increasing application rates of FGD. Leachates contained higher concentrations of B, with small increases in Se and As. Flue gas desulfurization residue application caused an increase in total B, As, Mo, Se, and extractable Ca in the soil, but decreased Mn and Zn. Using FGD residues could have beneficial effects on crop establishment without detrimental effects on soil or leachate quality, at an optimum rate of approximately 2.5%. This material could alleviate surface acidity, and B and Mo deficiencies in plants. PMID- 11401256 TI - Selenium distribution and fluxes in intertidal wetlands, San Francisco Bay, California. AB - Selenium (Se) concentrations exceeding ecological guidelines for sediments and suspended particulate matter (SPM) have been observed in the northern reach of the San Francisco Bay estuary. Longterm availability of elevated Se in wetland sediments depends in part on the fluxes controlling Se distribution. The relative contribution of sedimentary vs. post-depositional Se fluxes in two San Francisco Bay intertidal wetlands was estimated. Selenium concentrations on surface wetland sediments were compared with levels on SPM, and with previously established background levels in San Francisco Bay sediments. Sediment Se fluxes to the wetlands were measured directly using sediment traps. Although dissolved Se concentrations are higher than particulate Se concentrations in San Francisco Bay water, sediment input into the system provides the major flux of Se. Strong correlation between Se and C on SPM (r2 = 0.81) indicates the importance of organic particulate deposition. Dependence on sediment texture was qualitatively established by measuring Se on particle-size separates. Normalization to Al showed that 65% of Se spatial variability is related to sediment texture. Selenium is further enriched in the marsh via post-depositional inputs, probably due to in situ adsorption from overlying water and chemical reduction. According to sediment flux measurements, enrichment in the marsh is equivalent to 20 to 25% of the particulate Se flux, thereby defining the marsh as a Se sink. These findings highlight the need for more intensive monitoring of SPM as the major source of Se to intertidal wetlands. PMID- 11401257 TI - Monitoring nitrate leaching from submerged drains. AB - Monitoring nitrate N (NO3-N) leaching is important in order to judge the effect that agricultural practices have on the quality of ground water and surface water. Measuring drain discharge rates and NO3-N concentrations circumvents the problem of spatial variability encountered by other methods used to quantify NO3 N leaching in the field. A new flow-proportional drainage water sampling method for submerged drains has been developed to monitor NO3-N leaching. Both low and high discharge rates can be measured accurately, and are automatically compensated for fluctuations in ditch-water levels. The total amount of NO3-N leached was 10.6 kg N ha(-1) for a tile-drained silt-loam soil during the 114-d monitoring period. The NO3-N concentrations fluctuated between 5 mg L(-1) at deep ground water levels and 15 mg L(-1) at shallow levels, due to variations in water flow. A flow-proportional drainage water sampling method is required to measure NO3-N leaching accurately under these conditions. Errors of up to 43% may occur when NO3-N concentrations in the drainage water are only measured at intervals of 30 d and when the precipitation excess is used to estimate cumulative NO3-N leaching. Measurements of NO3-N concentrations in ground water cannot be used to accurately estimate NO3-N leaching in drained soils. PMID- 11401258 TI - Off-site movement of endosulfan from irrigated cotton in New South Wales. AB - The fate and transport of endosulfan (6,7,8,9,10,10-hexachloro-1,5, 5a,6,9,9a hexahydro-6,9-methano-2,4,3-benzodioxathiepin 3-oxide) applied to cotton (Gossypium hirsutum L.) fields were studied throughout three consecutive years on two selected locations in New South Wales (Australia). Rates of dissipation from foliage and soil, volatilization from the field, and transport of residues in irrigation and/or storm runoff waters were measured in order to estimate a total field balance. Dissipation of endosulfan from both foliage and soil is best explained by a two-phase process rather than by a first-order decay. Half-lives of total endosulfan toxic residues (alpha- and beta-endosulfan and the sulfate product) in the first phase were 1.6 d in foliage and 7.1 d in soil, and could be explained by the rapid volatilization of the parent isomers in the first 5 d (up to 70% of endosulfan volatilizes). In the second phase, half-lives were 9.5 d in foliage and 82 d in soil, mostly due to the persistence of the sulfate product. Concentration of endosulfan residues in runoff water varied from 45 to 2.5 microg L(-1) depending on the residue levels present on field soil at the time of the irrigation or storm events. These in turn are related to the total amounts applied, the cotton canopy cover at application, and the time since last spraying. Most of the endosulfan in runoff was found in the water phase (80%), suggesting it was bound to colloidal matter. Total endosulfan residues in runoff for a whole season accounted for no more than 2% of the pesticide applied on field. PMID- 11401259 TI - Spray drift of pesticides arising from aerial application in cotton. AB - This paper presents results from field studies carried out during the 1993-1998 Australian cotton (Gossypium hirsutum L.) seasons to monitor off-target droplet movement of endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9 methano-2,4,3-benzodioxathiepin 3-oxide) insecticide applied to a commercial cotton crop. Averaged over a wide range of conditions, off-target deposition 500 m downwind of the field boundary was approximately 2% of the field-applied rate with oil-based applications and 1% with water-based applications. Mean airborne drift values recorded 100 m downwind of a single flight line were a third as much with water-based application compared with oil-based application. Calculations using a Gaussian diffusion model and the U.S. Spray Drift Task Force AgDRIFT model produced downwind drift profiles that compared favorably with experimental data. Both models and data indicate that by adopting large droplet placement (LDP) application methods and incorporating crop buffer distances, spray drift can be effectively managed. PMID- 11401260 TI - Simulating endosulfan transport in runoff from cotton fields in Australia with the GLEAMS model. AB - Endosulfan (6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9methano-2,4,3 benzodioxathiepin 3-oxide), a pesticide that is highly toxic to aquatic organisms, is widely used in the cotton (Gossypium hirsutum L.) industry in Australia and is a risk to the downstream riverine environment. We used the GLEAMS model to evaluate the effectiveness of a range of management scenarios aimed at minimizing endosulfan transport in runoff at the field scale. The field management scenarios simulated were (i) Conventional, bare soil at the beginning of the cotton season and seven irrigations per season; (ii) Improved Irrigation, irrigation amounts reduced and frequency increased to reduce runoff from excess irrigation; (iii) Dryland, no irrigation; (iv) Stubble Retained, increased soil cover created by retaining residue from the previous crop or a specially planted winter cover crop; and (v) Reduced Sprays, a fewer number of sprays. Stubble Retained was the most effective scenario for minimizing endosulfan transport because infiltration was increased and erosion reduced, and the stubble intercepted and neutralized a proportion of the applied endosulfan. Reducing excess irrigation reduced annual export rates by 80 to 90%, but transport in larger storm events was still high. Reducing the number of pesticide applications only reduced transport when three or fewer sprays were applied. We conclude that endosulfan transport from cotton farms can be minimized with a combination of field management practices that reduce excess irrigation and concentration of pesticide on the soil at any point in time; however, discharges, probably with endosulfan concentrations exceeding guideline values, will still occur in storm events. PMID- 11401261 TI - Endosulfan transport: I. Integrative assessment of airborne and waterborne pathways. AB - To reduce endosulfan (C9H6O3Cl6S; 6,7,8,9,10,10-hexachloro-1,5, 5a,6,9,9a hexahydro-6,9-methano-2,4,3-benzodioxathiepin 3-oxide) contamination in rivers and waterways, it is important to know the relative significances of airborne transport pathways (including spray drift, vapor transport, and dust transport) and waterborne transport pathways (including overland and stream runoff). This work uses an integrated modeling approach to assess the absolute and relative contributions of these pathways to riverine endosulfan concentrations. The modeling framework involves two parts: a set of simple models for each transport pathway, and a model for the physical and chemical processes acting on endosulfan in river water. An averaging process is used to calculate the effects of transport pathways at the regional scale. The results show that spray drift, vapor transport, and runoff are all significant pathways. Dust transport is found to be insignificant. Spray drift and vapor transport both contribute low-level but nearly continuous inputs to the riverine endosulfan load during spraying season in a large cotton (Gossypium hirsutum L.)-growing area, whereas runoff provides occasional but higher inputs. These findings are supported by broad agreement between model predictions and observed typical riverine endosulfan concentrations in two rivers. PMID- 11401262 TI - Endosulfan transport: II. Modeling airborne dispersal and deposition by spray and vapor. AB - Endosulfan (C9H6O3Cl6S; 6,7,8,9,10,10-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9 methano-2,4,3-benzodioxathiepin 3-oxide) and other agricultural chemicals can be transported from farms to rivers by several airborne pathways including spray drift and vapor transport. This paper describes a modeling framework for quantifying both of these airborne pathways, consisting of components describing the source, dispersion, and deposition phases of each pathway. Throughout, the framework uses economical descriptions consistent with the need to capture the major physical processes. The dispersion of spray and vapor is described by similarity and mass-conservation principles approximated by Gaussian solutions. Deposition of particles to vegetation is described by a single-layer model incorporating contributions from settling, impaction, and Brownian diffusion. Vapor deposition to water surfaces is described by a simple kinetic formulation dependent on an exchange velocity. All model components are tested against available field and laboratory data. The models, and the measurements used for comparisons, both demonstrate that spray drift and vapor transport are significant pathways. The broader context, described in another paper, is an integrative assessment of all transport pathways (both airborne and waterborne) contributing to endosulfan transport from farms to rivers. PMID- 11401263 TI - Profenofos residues in wild fish from cotton-growing areas of New South Wales, Australia. AB - The organophosphorus (OP) pesticide profenofos (O-4-bromo-2-chlorophenyl O-ethyl S-propyl phosphorothioate) is used heavily in cotton-growing areas of eastern Australia toward the end of the growing season. European carp (Cyprinus carpio), bony bream (Nematalosa erebi), and mosquitofish (Gambusia holbrooki) were collected from the cotton-growing areas around Wee Waa, New South Wales, to determine the relationship between profenofos residues and acetyl-cholinesterase (AChE) activity in wild fish. Profenofos concentrations in water, sediment, and fish tissue reflected its general level of use; levels in March 1994 were significantly higher than in 1993 and generally decreased in May, 6 wk after cessation of spraying. Residues in carp and bony bream generally correlated with concentrations in water and sediment, although residues in fish tend to persist longer at some sites. Acetylcholinesterase inhibition was a useful indicator of profenofos exposure within a season, particularly if linked with residue measurements. Bony bream and gravid female mosquitofish recovered AChE levels more slowly than carp or nongravid mosquitofish. Recovery in creeks was generally more rapid than in lagoons. PMID- 11401264 TI - Fate and toxicity of endosulfan in Namoi River water and bottom sediment. AB - Endosulfan (6,7,8,9,10,10,-hexachloro-1,5,5a,6,9,9a-hexahydro-6,9-methano-2,4,3 benzodioxathiepine-3-oxide) sorption (standardized to 1% total organic carbon and dry weight) was significantly (P < 0.05) more concentrated on the large (>63 microm) particle fraction compared with smaller size fractions (<5 microm and 5 24 microm) of bottom sediments from the Namoi River, Australia. Following completion of the particle size fractionation (6 to 12 wk) and a sediment toxicity assessment (2 wk), the sediments showed large decreases in concentrations of alpha-endosulfan that coincided with an increase in endosulfan sulfate concentrations and minimal changes in beta-endosulfan concentrations. In the Namoi River, similar patterns were observed in the composition of total endosulfan in monthly measurements of bottom sediments and in passive samplers placed in the water column following runoff from cotton (Gossypium hirsutum L.) fields. The toxicity of endosulfan sulfate in river water indicated by the nymphs of the epibenthic mayfly Jappa kutera, was more persistent than the alpha- and beta-endosulfan parent isomers due to its longer half-life. This suggests that endosulfan sulfate would contribute most to previously observed changes in population densities of aquatic biota. Measured concentrations of total endosulfan in river water of up to 4 microg L(-1) following storm runoff, exceed the range of the 96-h median lethal concentration (LC50) values in river water for both alpha-endosulfan (LC50 = 0.7 microg L(-1); 95% confidence interval [CI] = 0.5 to 1.1) and endosulfan sulfate (LC50 = 1.2 microg L(-1); 95% CI = 0.4 to 3.3). In contrast, the 10-d LC50 value for total endosulfan in the sediment toxicity test (LC50 = 162 microg kg(-1); 95% CI = 120 to 218 microg kg(-1)) was more than threefold higher than the highest measured concentration of total endosulfan in field samples of bottom sediment (48 microg kg(-1)). This suggests that pulse exposures of endosulfan in the water column following storm runoff may be more acutely toxic to riverine biota than in contaminated bottom sediment. PMID- 11401265 TI - Olfactory response to mushroom composting emissions as a function of chemical concentration. AB - Odor pollution is a major problem facing mushroom [Agaricus bisporus (Lange) Imbach] compost production. Techniques for quantifying mushroom composting odors are needed to assess the effectiveness of odor control measures. Odor samples were obtained in nalophane bags from 11 mushroom composting sites. Samples were collected 0.2 m downwind from the pre-wetting heaps (aerated or unaerated) of raw composting ingredients (wheat straw, poultry and horse manures, and gypsum) and subsequent Phase I composting windrows or aerated tunnels. The odor concentrations (OCs) of the samples were assessed using serial dilution olfactometry and the chemical composition of the samples was determined using gas chromatography-mass spectrometry (GC-MS), both 24 h after sampling. Gas detector tubes were used for on-site measurement of gaseous compounds. Odorants that exceeded their published olfactory detection thresholds by the greatest order of magnitude, in decreasing order, were: H2S, dimethyl sulfide (DMS), butanoic acid, methanethiol, and trimethylamine. Concentrations of NH3 were not significantly correlated with OC, and they were not significantly affected by the use of aeration. Aeration reduced the OC and the combined H2S + DMS concentrations by 87 and 92%, respectively. There was a very close correlation (r = 0.948, P < 0.001) between the OC of bag samples and the combined H2S + DMS concentrations, measured on-site with detector tubes. This relationship was unaffected by the NH3 concentration or the type of compost: aerated or unaerated, pre-wet or Phase I, poultry manure-based or horse and poultry manure-based compost. Prediction of the OC will enable rapid and low-cost identification of odor sources on mushroom composting sites. PMID- 11401266 TI - Atmospheric movements of lindane (gamma-hexachlorocyclohexane) from canola fields planted with treated seed. AB - Lindane (gamma-hexachlorocyclohexane [gamma-HCH]) is used as an insecticide in many countries. Concentrations of gamma-HCH have been found in air, water, soil, snow, and tissue samples throughout the world and concerns have been raised for its potential effects on human and ecosystem health. In Canada, gamma-HCH is primarily used as a treatment on canola (Brassica napus L) seed with an estimated 455.3 Mg applied in 1997 and 510.4 Mg in 1998. The purpose of this study was to measure gamma-HCH volatilization from fields planted with treated canola seed. Atmospheric dry and wet deposition and soil samples were collected for two growing seasons (1997 and 1998) from a canola field planted with treated seed. Atmospheric concentrations as high as 16.1 and 7.4 ng m(-3) were measured at 1 m above the canola field compared with maximum concentrations of 2.9 and 2.7 ng m( 3) measured above a grass field located 2 km away (1997 and 1998, respectively). On the basis of measurements made in this study it was estimated that between 12 and 30% of the gamma-HCH applied as canola seed treatment may volatilize and be released to the atmosphere. This would create an atmospheric loading of 66.4 to 188.8 Mg for the 6-wk period following planting, estimated from the quantity of seed sown on the Canadian prairies in 1998. Dry deposition rates and rain concentrations as high as 2,203 ng m(-2) d(-1) and 170 ng L(-1) were measured adjacent to the canola field. PMID- 11401267 TI - A micrometeorological technique to monitor total hydrocarbon emissions from landfarms to the atmosphere. AB - Landfarming is used to treat petroleum hydrocarbon-contaminated soils and a variety of waste streams from industrial operations. Wastes are applied to a soil surface and indigenous soil microorganisms utilize the hydrocarbons in the applied waste as a carbon source for metabolism, thereby biodegrading the applied material. Concerns have been expressed that abiotic losses, such as volatilization, play a significant role in hydrocarbon reduction within the soil. To assist in better defining atmospheric releases of total hydrocarbons from landfarms treating petroleum hydrocarbons, a flux gradient micrometeorological approach was developed and integrated with a custom-built total hydrocarbon detector, and a novel air sampling system and averaging algorithm. The micrometeorological technique offers unobtrusive spatially averaged real-time continuous measurements, thereby providing a time history of emissions. This provides opportunities to investigate mechanisms controlling emissions and to evaluate landfarm management strategies. The versatility of the technique is illustrated through measurements performed at a remote landfarm used to treat diesel fuel-contaminated soil in northern Ontario and during routine operations at two active refinery landfarms in southwestern Ontario. PMID- 11401268 TI - Effect of land use on methane flux from soil. AB - The precise effects of natural and disturbed terrestrial systems on the atmospheric CH4 pool are uncertain. This study was conducted to quantify and compare CH4 fluxes from a variety of ecosystems in central Iowa. We investigated agricultural systems under different management practices, a hardwood forest site, native and restored prairies, and a municipal landfill. Flux measurements were obtained using a closed-chamber method, and measurements were compiled by sampling over the 1993 and 1994 growing seasons. In 1993, most of the agricultural sites were net CH4 producers with cumulative CH4 fluxes ranging from -0.02 to 3.19 g m(-2) over the 258-d sampling season, while the natural ecosystems were net CH4 consumers, with cumulative seasonal fluxes ranging from 0.27 to -0.07 g m-2 258 d(-1). In 1994, only the landfill and the agricultural site treated with broadcast liquid swine manure (LSM) were net CH4 producers, while the remainder of the natural and agricultural ecosystems were net CH4 consumers, with mean seasonal flux rates ranging from -0.43 to -0.008 g m(-2) 271 d(-1). We hypothesize that the differences in CH4 fluxes between the two years are due to differences in rainfall. To illustrate the integration between land use and CH4 flux, we computed an area-weighted soil CH4 flux for the state of Iowa. Our calculations yielded a net average soil CH4 flux of 139,000 Mg CH4 for 1993 and 1994. PMID- 11401269 TI - Pesticide risk reduction on crops in the Province of Ontario. AB - We analyzed the changes in pesticide use and risk in the Province of Ontario, Canada, from 1973 to 1998 to monitor the success of Food Systems 2002, a program to reduce pesticide use by 50%. Pesticide risk was calculated by multiplying the amount of pesticide used (kilograms of active ingredient) by the Environmental Impact Quotient (EIQ), a score for the potential risk of pesticides to farmworkers, consumers, and the environment. Pesticide use increased by 46% from 1973 to 1983. From 1983, the baseline year for Food Systems 2002, to 1998, pesticide use decreased by 38.5% and risk declined 39.5%. The reductions in pesticide use and risk were primarily on corn (Zea mays L.) and tobacco (Nicotiana tabacum L.), the crops with the highest pesticide use in 1983. Total pesticide use on soybean [Glycine max (L.) Merr.] did not change, but the mean application rate (kg ha(-1)) decreased by 57%. Corn and soybean account for 65% of pesticide use, but have a relatively low pesticide use and risk per hectare and per tonne of production. Total pesticide use on tobacco, fruits, and vegetables was lower than on corn or soybean, but the pesticide use and risk per hectare were much higher. Small reductions in pesticide use on corn and soybean may allow a 50% reduction in pesticide use, but greater reductions in risk can be achieved by reducing the use of "high risk" pesticides on fruit and vegetables. PMID- 11401270 TI - Spray deposition of two insecticides into surface waters in a South African orchard area. AB - Drift from pesticide spray application can result in contamination of nontarget environments such as surface waters. Azinphos-methyl (AZI) and endosulfan (END) deposition in containers of water was studied in fruit orchards in the Western Cape, South Africa. Additionally, attention was given to the contamination in farm streams, as well as to the resulting contamination of the subsequent main channel (Lourens River) approx. 25 km downstream of the tributary stream inlets. Spray deposit decreased with increasing distance downwind and ranged from 4.7 mg m(-2) within the target area to 0.2 mg m(-2) at 15 m downwind (AZI). Measured in stream concentrations of both pesticides compared well with theoretical values calculated from deposition data for the respective distances. Furthermore, they were in the range of values predicted by an exposure assessment based on 95th percentile values for basic drift deposition (German Federal Biological Research Centre for Agriculture and Forestry [BBA] and USEPA). Pesticide deposition in the tributaries was followed by a measurable increase of contamination in the Lourens River. Mortality of midges (Chironomus spp.) exposed for 24 h to samples obtained from the AZI trials decreased with decreasing concentrations (estimated LC50 from field samples = 10 microg L(-1) AZI; lethal distance: LD50 = 13 m). Mortality in the tributary samples averaged 11% (0.5-1.7 microg L(-1) AZI), while no mortality was discernible in the Lourens River samples (0.041 microg L(-1)). The sublethal endpoint failure to form tubes from the glass beads provided was significantly increased at all sites in comparison with the control (analysis of variance [ANOVA], Fisher's protected least significant difference [PLSD], p < 0.01). PMID- 11401271 TI - EPIC tile flow and nitrate loss predictions for three Minnesota cropping systems. AB - Subsurface tile drains are a key source of nitrate N (NO3-N) losses to streams in parts of the north central USA. In this study, the Erosion Productivity Impact Calculator (EPIC) model was evaluated by comparing measured vs. predicted tile flow, tile NO3-N loss, soil profile residual NO3-N, crop N uptake, and yield, using 4 yr of data collected at a site near Lamberton, MN, for three crop rotations: continuous corn (Zea mays L.) or CC, corn-soybean [Glycine max (L.) Merr.] or CS, and continuous alfalfa (Medicago sativa L.) or CA. Initially, EPIC was run using standard Soil Conservation Service (SCS) runoff curve numbers (CN2) for CC and CS; monthly variations were accurately tracked for tile flow (r2 = 0.86 and 0.90) and NO3-N loss (r2 = 0.69 and 0.52). However, average annual CC and CS tile flows were underpredicted by -32 and -34%, and corresponding annual NO3-N losses were underpredicted by -11 and -52%. Predicted average annual tile flows and NO3-N losses generally improved following calibration of the CN2; tile flow underpredictions were -9 and - 12%, whereas NO3-N losses were 0.6 and -54%. Adjusting a N parameter further improved predicted CS NO3-N losses. Predicted monthly tile flows and NO3-N losses for the CA simulation compared poorly with observed values (r2 values of 0.27 and 0.19); the annual drainage volumes and N losses were of similar magnitude to those measured. Overall, EPIC replicated the relative impacts of the three cropping systems on N fate. PMID- 11401272 TI - Major herbicides in ground water: results from the National Water-Quality Assessment. AB - To improve understanding of the factors affecting pesticide occurrence in ground water, patterns of detection were examined for selected herbicides, based primarily on results from the National Water-Quality Assessment (NAWQA) program. The NAWQA data were derived from 2,227 sites (wells and springs) sampled in 20 major hydrologic basins across the USA from 1993 to 1995. Results are presented for six high-use herbicides--atrazine (2-chloro-4-ethylamino-6-isopropylamino-s triazine), cyanazine (2-[4-chloro-6-ethylamino-1,3,5triazin-2-yl]amino]-2 methylpropionitrile), simazine (2-chloro-4,6-bis-[ethylamino]-s-triazine), alachlor (2-chloro-N-[2,6-diethylphenyl]-N-[methoxymethyl]acetamide), acetochlor (2-chloro-N-[ethoxymethyl]-N-[2-ethyl-6-methylphenyl]acetamide), and metolachlor (2-chloro-N-[2-ethyl-6-methylphenyl]-N-[2-methoxylethyl]acetamide)--as well as for prometon (2,4-bis[isopropylamino]-6-methoxy-s-triazine), a nonagricultural herbicide detected frequently during the study. Concentrations were <1 microg L( 1) at 98% of the sites with detections, but exceeded drinking-water criteria (for atrazine) at two sites. In urban areas, frequencies of detection (at or above 0.01 microg L(-1)) of atrazine, cyanazine, simazine, alachlor, and metolachlor in shallow ground water were positively correlated with their nonagricultural use nationwide (P < 0.05). Among different agricultural areas, frequencies of detection were positively correlated with nearby agricultural use for atrazine, cyanazine, alachlor, and metolachlor, but not simazine. Multivariate analysis demonstrated that for these five herbicides, frequencies of detection beneath agricultural areas were positively correlated with their agricultural use and persistence in aerobic soil. Acetochlor, an agricultural herbicide first registered in 1994 for use in the USA, was detected in shallow ground water by 1995, consistent with previous field-scale studies indicating that some pesticides may be detected in ground water within 1 yr following application. The NAWQA results agreed closely with those from other multistate studies with similar designs. PMID- 11401273 TI - Adsorption of pesticides onto quartz, calcite, kaolinite, and alpha-alumina. AB - The fate of pesticides in aquifers is influenced by the small but not insignificant adsorption of pesticides to mineral surfaces. Batch experiments with five pesticides and four minerals were conducted to quantify the contributions to adsorption from different mineral surfaces and compare adsorption characteristics of selected pesticides. Investigated mineral phases included quartz, calcite, kaolinite, and alpha-alumina. Selected pesticides comprised atrazine (6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine), isoproturon [3-(4-isopropylphenyl)-1,1-dimethylurea)], mecoprop [(RS)-2-(4-chloro 2-methylphenoxy)propionic acid], 2,4-D (2,4-dichlorophenoxyacetic acid), and bentazone [3-isopropyl-1H-2,1,3-benzothiadiazin-4-(3H)-one 2,2-dioxide]. Specific surface area and mineral surface charge proved to be important for the adsorption of these pesticides. Detectable adsorption of the anionic pesticides (mecoprop, 2,4-D, and bentazone) was only measured when positive sites were present on the mineral surface. However, when CaCl2 was added as an electrolyte, a detectable adsorption of mecoprop and 2,4-D was also measured on kaolinite (which exhibits a negative surface charge), probably due to formation of Ca-pesticide--surface complexes. Adsorption of the uncharged pesticides (atrazine and isoproturon) was detected only on kaolinite. The lack of adsorption on alpha-alumina indicates that the uncharged pesticides have a greater affinity for the silanol surface sites (=SiOH) than for the aluminol surface sites (=AlOH) in kaolinite. No measurable effect of ionic strength was found for the uncharged pesticides. The results indicate that quartz and calcite play a smaller role than clay minerals. PMID- 11401274 TI - Atrazine, isoproturon, mecoprop, 2,4-D, and bentazone adsorption onto iron oxides. AB - Iron oxides are important components influencing the adsorption of various inorganic and organic compounds in soils and sediments. In this study the adsorption on iron oxides of nonionic and ionic pesticides was determined as a function of solution pH, ionic strength, and pesticide concentration. The investigated iron oxides included two-line ferrihydrite, goethite, and lepidocrocite. Selected pesticides comprised atrazine (6-chloro-N2-ethyl-N4 isopropyl-1,3,5-triazine-2,4-diamine), isoproturon [3-(4-isopropylphenyl)-1,1 dimethylurea)], mecoprop [(RS)-2-(4-chloro-2-methylphenoxy)propionic acid], 2,4-D (2,4-dichlorophenoxyacetic acid), and bentazone [3-isopropyl-1H-2,1,3 benzothiadiazin-4(3H)-one 2,2-dioxide]. The adsorption of the nonionic pesticides (atrazine and isoproturon) was insignificant, whereas the adsorption of the acidic pesticides (mecoprop, 2,4-D, and bentazone) was significant on all investigated iron oxides. The adsorption capacity increased with decreasing pH, with maximum adsorption reached close to the pKa values. The addition of CaCl2 in concentrations from 0.0025 to 0.01 M caused the adsorption capacity to diminish. The adsorption of bentazone was significantly lower than the adsorption of mecoprop and 2,4-D, illustrating the importance of a carboxyl group in the pesticide structure. The adsorption capacity on the iron oxides increased in the order: lepidocrocite < goethite < two-line ferrihydrite. The maximum adsorption capacities of meco-prop and 2,4-D on goethite were found to be equivalent to the site density of singly coordinated hydroxyl groups on the faces of the dominant (110) form, suggesting that singly coordinated hydroxyl groups are responsible for adsorption. Differences in adsorption capacities between iron oxides can be explained by differences in the surface site density of singly coordinated hydroxyl groups. The maximum measured adsorption capacity of mecoprop on two-line ferrihydrite was equivalent to 0.2 mol/mol Fe. PMID- 11401275 TI - Plant uptake of cadmium-109 and zinc-65 at different temperature and organic matter levels. AB - The uptake of 109Cd and 65Zn and their stable isotopes by ryegrass (Lolium multiflorum Lam.), grown on two different soil types, was investigated in climatically controlled growth chambers at 9 and 21 degrees C. The soils were treated with 0 and 4% organic matter (pig [Sus scrofa] manure) and spiked with 109Cd and 65Zn before sowing. The organic matter addition resulted in increased uptake of the 109Cd, Cd, and Zn by ryegrass, but the uptake of 65Zn was decreased. The latter effect was ascribed to isotopic dilution of 65Zn as the amount of stable Zn in the plant tissues increased with the organic matter addition. The effect of temperature was more pronounced than that of organic matter addition, and the uptake of both 109Cd and 65Zn and their stable isotopes was higher in ryegrass grown at 21 degrees C than that grown at 9 degrees C. Results from fractionation and speciation analysis of soil cadmium and zinc were correlated with plant uptake, and there was a good consistency between observed plant uptake and the physico-chemical forms of cadmium and zinc in soil and soil solution presumed to be plant available. PMID- 11401276 TI - Effect of dissolved organic matter from sludge and sludge compost on soil copper sorption. AB - Interaction of Cu with dissolved organic matter (DOM) is an important physicochemical process affecting Cu mobility in soils. The aim of this study was to investigate the effects of DOM from anaerobically digested dewatered sludge and sludge compost on the sorption of Cu on an acidic sandy loam and a calcareous clay loam. In the presence of DOM, Cu sorption capacity decreased markedly for both soils, especially for the calcareous soil. The Cu sorption isotherms could be well described by the Freundlich equation (r2 = 0.99), and the binding intensity parameter of soils in the presence of sludge DOM was lower than compost DOM. An increase in DOM concentration significantly reduced the sorption of Cu by both soils. Within the Cu and DOM concentration range studied, the decrease in Cu sorption caused by sludge DOM was consistently greater than that of compost DOM. This might be attributed to the greater amount of hydrophobic fraction of DOM in the compost. Moreover, the reduction of Cu sorption caused by DOM was more obvious in the soil with higher pH. In addition, the sorption of Cu increased with an increase in pH for both soils without the addition of DOM, while Cu sorption in the presence of DOM was unexpectedly decreased with an increase in pH at a pH >6.8. This implied that DOM produced by sludge or other C-enriched organic wastes heavily applied on calcareous soils might facilitate the leaching loss of Cu because of the formation of soluble DOM-metal complexes. PMID- 11401277 TI - Photosynthesis, chlorophyll integrity, and spectral reflectance in lichens exposed to air pollution. AB - The major objective of the present study was to identify the relationship of physiologial parameters of the photosynthetic system with the elemental content of the lichen Ramalina lacera (With.) J.R. Laund. Thalli of R. lacera were collected in an unpolluted site and transplanted in a national park and an industrial region in Israel for 8 mo. Analyses of photosynthetic activity, chlorophyll integrity, spectral reflectance, and amount of 11 metals were performed after this period of exposure. The normalized difference vegetation index (NDVI), indicative of the spectral reflectance response of the thallus, correlated with photosynthetic rate and chlorophyll and K content and correlated inversely with amounts of Ba, Cr, Cu, and Ni. The NDVI appears to enable the detection of early signs of pollutant-induced stress before changes in other physiological parameters become apparent. Elevated amounts of Cr, Cu, Fe, Mn, Ni, and Zn in lichens transplanted to an industrial area and the correlation of Mn and Ni, Mn and V, Ni and V, Fe and Mn, Fe and V, and Fe and Zn point for the greater part to metal processing in a steel smelter. Correlations of Cr and Ni, Cu and Ni, Zn and Cu, Cu and Mn, and Zn and Ni could be related to metal processing in the industrial area but indicate also vehicular activity as a possible originator. PMID- 11401278 TI - Using rank-order geostatistics for spatial interpolation of highly skewed data in a heavy-metal contaminated site. AB - The spatial distribution of a pollutant in contaminated soils is usually highly skewed. As a result, the sample variogram often differs considerably from its regional counterpart and the geostatistical interpolation is hindered. In this study, rank-order geostatistics with standardized rank transformation was used for the spatial interpolation of pollutants with a highly skewed distribution in contaminated soils when commonly used nonlinear methods, such as logarithmic and normal-scored transformations, are not suitable. A real data set of soil Cd concentrations with great variation and high skewness in a contaminated site of Taiwan was used for illustration. The spatial dependence of ranks transformed from Cd concentrations was identified and kriging estimation was readily performed in the standardized-rank space. The estimated standardized rank was back-transformed into the concentration space using the middle point model within a standardized-rank interval of the empirical distribution function (EDF). The spatial distribution of Cd concentrations was then obtained. The probability of Cd concentration being higher than a given cutoff value also can be estimated by using the estimated distribution of standardized ranks. The contour maps of Cd concentrations and the probabilities of Cd concentrations being higher than the cutoff value can be simultaneously used for delineation of hazardous areas of contaminated soils. PMID- 11401279 TI - Adsorption of cadmium on biosolids-amended soils. AB - Debate exists over the biosolid phase (organic or inorganic) responsible for the reduction in phytoavailable Cd in soils amended with biosolids as compared with soils amended with inorganic salts. To test the importance of these two phases, adsorption isotherms were developed for soil samples (nine biosolids-amended soils and their five companion controls) and two biosolids samples from five experimental sites with documented histories of biosolids application. Subsamples were treated with 0.7 M NaClO to remove organic carbon. Cadmium nitrate was added to both moist soil samples and their soil inorganic fractions (SIF) in a 0.01 M Ca(NO3)2 solution at three pH levels (6.5, 5.5, and 4.5), and equilibrated at 22 +/- 1 degrees C for at least 48 h. Isotherms of Cd adsorption for biosolids amended soil were intermediate to the control soil and biosolids. Decreasing pH did not remove the difference between these isotherms, although adsorption of Cd decreased with decreasing pH level. Organic matter removal reduced Cd adsorption on all soils but had little influence on the observed difference between biosolids-amended and control soils. Thus, increased adsorption associated with biosolids application was not limited to the organic matter addition from biosolids; rather, the biosolids application also altered the adsorptive properties of the SIF. The greater affinity of the inorganic fraction of biosolids-amended soils to adsorb Cd suggests that the increased retention of Cd on biosolids-amended soils is independent of the added organic matter and of a persistent nature. PMID- 11401280 TI - Accumulation, distribution, and toxicity of copper in sediments of catfish ponds receiving periodic copper sulfate applications. AB - Copper sulfate (CuSO4) is applied periodically to commercial channel catfish (Ictalurus panctatus) ponds as an algicide or parasiticide. Current understanding of the chemistry of copper in soil-water systems suggests that copper may accumulate in pond sediments, although the forms and potential bioavailability of copper in catfish pond sediments are not known. This study investigated the accumulation and distribution of copper in the sediment of catfish ponds receiving periodic additions of CuSO4.5H2O. All ponds were constructed in Sharkey (very-fine, smectitic, thermic Chromic Epiaquert) soil. Nine 0.40-ha ponds received 59 applications of 2.27 kg CuSO4.5H2O per application per pond over 3 yr; no CuSO4.5H2O applications were made to nine additional ponds. Total Cu concentration in the sediments of CuSO4.5H2O-amended catfish ponds (172.5 mg kg( 1)) was four to five times higher than that in the sediments of nonamended ponds (36.1 mg kg(-1)). Copper accumulated in catfish pond sediments at a rate of 41 microg kg(-1) dry sediment for each 1 kg ha(-1) of CuSO4. 5H2O applied to ponds. Copper in the sediments of amended ponds was mainly in the organic matter-bound (30.7%), carbonate-bound (31.8%), and amorphous iron oxide-bound (22.1%) fractions with a considerable fraction (3.4%; 3 to 8 mg kg(-1)) in soluble and exchangeable fractions. This indicates that Cu accumulates differentially in various fractions, with proportionally greater initial accumulation in potentially bioavailable forms. However, toxicity bioassays with amphipods (Hyallela azteca) and common cattail (Typha latifolia L.) indicated that the effect of exposure to amended or nonamended pond sediments was not different. PMID- 11401281 TI - Elevated lead and zinc contents in remote alpine soils of the Swiss National Park. AB - Weathering of bedrock and pedogenic processes can result in elevated heavy metal concentrations in the soil. Small-scale variations in bedrock composition can therefore cause local variations in the metal content of the soil. Such a case was found in the remote alpine area of the Swiss National Park. Soil profiles were sampled at an altitude of about 2,400 m, representing soils developed above different bedrocks. The concentration of lead in the profiles was found to be strongly dependent on the metal content in the bedrock underlying the soil and was strongly enriched in the top 10 cm. The dolomitic bedrock in the study area contains elevated lead concentrations compared with other dolomites. Dissolution of dolomite and accumulation of weathering residues during soil formation resulted in high lead concentrations throughout the soil profile. The enrichment of lead in the topsoil, however, is largely attributed to atmospheric input. The isotopic signature of the lead clearly indicates that it is mainly of natural origin and that atmospheric deposition of anthropogenic lead contributed to about 20 to 40% to the lead concentration in the topsoil on the bedrock with elevated lead concentrations. In the soils on bedrock with normal lead concentrations, the anthropogenic contribution is estimated to be about 75%. Also, zinc was very strongly enriched in the topsoil. This enrichment was closely correlated with the organic matter distribution in the profiles, suggesting that recycling through plant uptake and litter deposition was a dominant process in the long-term retention of this metal in the soil. PMID- 11401282 TI - Copper and calcium transport through an unsaturated soil column. AB - To determine the relative importance of the physical and chemical factors that influence the movement of heavy metals through soils, leaching experiments were carried out under conditions of constant molarity during unsaturated steady-state water flow through a Manawatu fine sandy loam (a Dystric Fluventic Eutochrept). The movement and exchange of copper was studied in a binary Cu-Ca system. The movement of the associated anions, namely chloride and sulfate, was also monitored. The measurements were compared with predictions from the convection dispersion equation (CDE), linked with cation exchange theory. The agreement between the measured and predicted breakthrough of sulfate and copper was good. This indicates that copper retardation in the Manawatu soil is closely related to the cation exchange capacity, and that exchange between Ca and Cu is the main process of Cu retardation in the Manawatu soil. However, copper appeared slightly later in the effluent than predicted, indicating that non-exchange processes are also involved in copper transport. Measurements of suction cups could also be used to obtain the parameters for the CDE to describe sulfate movement through the soil. Time domain reflectometry (TDR) measurements of the bulk-soil electrical conductivity could be used to monitor the movement of both sulfate and copper. This indicates that TDR can also be used to monitor cation transport and exchange through the soil, provided the percolating solution causes a sufficient change in the electrical conductivity. PMID- 11401283 TI - Availability of arsenic, copper, lead, thallium, and zinc to various vegetables grown in slag-contaminated soils. AB - To anticipate a possible hazard resulting from the plant uptake of metals from slag-contaminated soils, it is useful to study whether vegetables exist that are able to mobilize a given metal in the slag to a larger proportion than in an uncontaminated control soil. For this purpose, we studied the soil to plant transfer of arsenic, copper, lead, thallium, and zinc by the vegetables bean (Phaseolus vulgaris L. 'dwarf bean Modus'), kohlrabi (Brassica oleracea var. gongylodes L.), mangold (Beta vulgaris var. macrorhiza ), lettuce (Lactuca sativa L. 'American gathering brown'), carrot (Daucus carota L. 'Rotin', 'Sperlings's'), and celery [Apium graveiolus var. dulce (Mill.) Pers.] from a control soil (Ap horizon of a Entisol) and from a contaminated soil (1:1 soil-slag mixtures). Two types of slags were used: an iron-rich residue from pyrite (FeS2) roasting and a residue from coal firing. The metal concentrations in the slags, soils, and plants were used to calculate for each metal and soil-slag mixture the plant-soil fractional concentration ratio (CRfractional,slag), that is, the concentration ratio of the metal that results only from the slag in the soil. With the exception of TI, the resulting values obtained for this quantity for As, Cu, Pb, and Zn and for all vegetables were significantly smaller than the corresponding plant-soil concentration ratios (CRcontrol soil) for the uncontaminated soil. The results demonstrate quantitatively that the ability of a plant to accumulate a given metal as observed for a control soil might not exist for a soil-slag mixture, and vice versa. PMID- 11401284 TI - Timing of phosphorus fertilizer application within an irrigation cycle for perennial pasture. AB - Irrigated pastures are significant contributors of phosphorus (P) to inland watercourses, with much of the P coming from applied fertilizer. It was hypothesized that the timing of P fertilizer application relative to irrigation regulates P concentrations in runoff and infiltrating water. To test this hypothesis, a two-by-two factorial experiment was conducted on twelve 8- x 30-m border-irrigated bays growing perennial pasture. Phosphorus fertilizer in the form of single superphosphate (44 kg P ha(-1)) was surface-broadcast onto the bays when the nominal change in soil water deficit reached 0 or 50 mm (U.S. Class A pan evaporation minus rainfall). Following fertilizer application, the bays were again irrigated when the nominal soil water deficit between fertilizing and the subsequent irrigation reached either 0 or 50 mm. The volume of water applied, runoff volume, and changes in soil water content were recorded for the three irrigations following fertilizer application. Total phosphorus (TP) and filtrable reactive phosphorus (FRP, <0.45 microm) concentrations in runoff and at depths of 0.1, 0.3, and 0.6 m in the soil were also measured. Soil water content at fertilizer application had less effect on P concentrations in runoff and soil water than the additional time between fertilizing and irrigating. By allowing a deficit of 50 mm between fertilizer application and irrigation, the average concentration of P in runoff and moving below a soil depth of 0.1 m was approximately halved. To maximize fertilizer use efficiency and minimize environmental effects, a delay should occur between applying P fertilizer and irrigating perennial pasture. PMID- 11401285 TI - Laboratory degradation studies of bentazone, dichlorprop, MCPA, and propiconazole in Norwegian soils. AB - Laboratory degradation studies were performed in Norwegian soils using two commercial formulations (Tilt and Triagran-P) containing either propiconazole alone or a combination of bentazone, dichlorprop, and MCPA. These soils included a fine sandy loam from Hole and a loam from Kroer, both of which are representative of Norwegian agricultural soils. The third soil was a highly decomposed organic material from the Froland forest. A fourth soil from the Skuterud watershed was used only for propiconazole degradation. After 84 d, less than 0.1% of the initial MCPA concentration remained in all three selected soils. For dichlorprop, the same results were found for the fine sandy loam and the organic-rich soil, but in the loam, 26% of the initial concentration remained. After 84 d, less than 0.1% of the initial concentration of bentazone remained in the organic-rich soil, but in the loam and the fine sandy loam 52 and 69% remained, respectively. Propiconazole was shown to be different from the other pesticides by its persistence. Amounts of initial concentration remaining varied from 40, 70, and 82% in the reference soils after 84 d for the organic-rich soil, fine sandy loam, and loam, respectively. The organic-rich soil showed the highest capacity to decompose all four pesticides. The results from the agricultural soils and the Skuterud watershed showed that the persistence of propiconazole was high. Pesticide degradation was approximated to first-order kinetics. Slow rates of degradation, where more than 50% of the pesticide remained in the soil after the 84-d duration of the experiment, did not fit well with first-order kinetics. PMID- 11401286 TI - Application of calorimetry to microbial biodegradation studies of agrochemicals in oxisols. AB - Calorimetry was used to monitor the inhibitory effect caused by the bipyridynium diquaternary salts paraquat, diquat, and phosphamidon on microbial activity in a Red Latosol soil (Oxisol). The thermal effect was recorded on samples composed of 1.50 g of soil, 6.0 mg of glucose, 6.0 mg of ammonium sulfate, and different masses of an inhibitor ranging from zero to 8.00 mg, under a controlled moisture content of 35%. Thermal effects of each pollutant on the degradation curves of glucose in the soil were compared. Increasing amounts of the inhibitor caused a decrease in the thermal effect from -2234 to -1987 kJ mol(-1) for paraquat, -1670 to -1306 kJ mol(-1) for diquat, and -2239 to -589 kJ mol(-1) for phosphamidon. The last xenobiotic agent caused a significant inhibitory effect on the microbial activity of the soil. The results of relative efficiency, eta = deltaH/deltaH', referring to the enthalpic value with (deltaH) and without (deltaH') agrochemical in the soil, exhibited a significant correlation. From this correlation obtained for the ranges 2.00 to 8.00, 1.30 to 8.00, and 1.20 to 5.80 mg of the agrochemicals paraquat, diquat, and phosphamidon, respectively, the following eta values were calculated: 0.993 to 0.894, 0.668 to 0.522, and 0.896 to 0.236, respectively, during the degradation of glucose in the soil. The largest relative efficiency for paraquat implies that this agrochemical can be metabolized by microbial activity. PMID- 11401287 TI - Assessment of the risk of phosphorus loading due to resuspended sediment. AB - Resuspension is a multiphase phenomenon where suspended solids encounter water layers differing in physico-chemical properties that affect the reactions of phosphorus (P). The role of resuspended sediment as a sink or source of dissolved P was determined in a laboratory study of P desorption-sorption equilibria. Gradual mixing was simulated using decreasing solid concentrations and varying environmental conditions (pH, redox, ionic strength). To describe the P exchange when the particles encounter dissimilar water layers, the extent of P sorption to or desorption from solids was expressed as a function of P concentration in the bath solutions. The equilibrium phosphorus concentration (EPC), at which there is no net P release from or retention to the particles, proved to be a suitable parameter for assessment of P load risk. Under oxic conditions at pH 7, commonly prevailing in lakes, the EPC values ranged from 11 to 27 microg P L(-1). The larger the water volume the suspended material was mixed with, the higher the P concentration, allowing desorption to occur. As for chemical factors affecting P mobilization, EPC followed the order: pH 7 < pH 7 anoxic < pH 9. A separate extraction experiment revealed that elevated pH enhanced P mobilization more as the concentration of solids decresed. The results demonstrate that high pH (a common characteristic in eutrophic lakes during summer), when linked with intensive resuspension, may markedly increase the internal P loading risk. As for the risk assessment, the quantification of the internal P loading would be improved by isotherm studies combined with field observations. PMID- 11401288 TI - Precipitation and river water chemistry of the Piracicaba River basin, southeast Brazil. AB - Annual precipitation and river water volumes and chemistry were measured from 1995 to 1998 in a mesoscale agricultural area of southeast Brazil. Precipitation was mildly acidic and solute concentrations were higher in the west than in the east of the basin. Combustion products from biomass burning, automobile exhaust, and industry typically accumulate in the atmosphere from March until October and are responsible for seasonal differences observed in precipitation chemistry. In river waters, the volume-weighted mean (VWM) concentrations of major solutes at 10 sites across the basin were generally lower at upriver than at downriver sampling sites for most solutes. Mass balances for major solutes indicate that, as a regional entity, the Piracicaba River basin was a net sink of H+, PO4(3-), and NH4+, and a net source of other solutes. The main stem of the Piracicaba River had a general increase in solute concentrations from upriver to downriver sampling sites. In contrast, NO3- and NH4+ concentrations increased in the mid reach sampling sites and decreased due to immobilization or utilization in the mid-reach reservoir, and there was denitrification immediately downriver of this reservoir. Compared with tributaries of the Chesapeake Bay estuary, the Piracicaba River is affected more by point-source inputs of raw sewage and industrial wastes than nonpoint agricultural runoff high in N and P. Inputs of N and C are responsible for a degradation of water quality at downriver sampling sites of the Piracicaba River drainage, and water quality could be considerably improved by augmenting sewage treatment. PMID- 11401289 TI - Herbicide transport to surface waters at field and watershed scales in a Mediterranean vineyard area. AB - The contamination of soil and runoff water by two herbicides, diuron [N'-(3,4 dichlorphenyl)-N,N-dimethylurea] and simazine (6-chloro-N,N'-diethyl-1,3,5 triazine-2,4-diamine), were monitored on two fields, one no-till and one tilled. Experiments were carried out in a 91.4-ha watershed in southern France during the 1997 growing season in order to understand the patterns of pesticide transport from field to watershed. The persistence of the herbicides in soil was prolonged due to the climatic conditions. At the field scale, annual herbicide loads were due to overland flow and amounted to 65.6 and 6.3 g ha(-1) of diuron for the no till and tilled field, respectively, and to 29.6 and 1.83 g ha(-1) of simazine. Maximum herbicide concentrations exceeded 580 microg L(-1) during the first storm event after application and decreased thereafter but remained for 8 mo above 0.1 microg L(-1). At the watershed outlet, estimated annual loads amounted to 4.12 g ha(-1) of diuron and 0.56 g ha(-1) of simazine. Among them, 96% of the losses in diuron and 83% of those in simazine were caused by the fast transmission through the network of ditches of the overland flow exiting the fields. For diuron, which was sprayed over most of the vineyards, its in-stream concentrations during storm flow were close to those at the outlet of the fields. The herbicide loads in baseflow were smaller than 0.2 g ha(-1). The patterns of the loads at the field and watershed scales suggested that a major part of the herbicides leaving the fields reinfiltrated to the ground water by seepage through the ditches, and was there degraded or adsorbed. PMID- 11401290 TI - Effects of alum and aluminum chloride on phosphorus runoff from swine manure. AB - Phosphorus (P) runoff from fields fertilized with swine (Sus scrofa domesticus) manure may contribute to eutrophication. The objective of this study was to evaluate the effect of aluminum sulfate (alum) and aluminum chloride applications to swine manure on P runoff from small plots cropped to tall fescue (Festuca arundinacea Shreb.). There were six treatments in this study: (i) unfertilized control plots, (ii) untreated manure, (iii) manure with alum at 215 mg Al L(-1), (iv) manure with aluminum chloride at 215 mg Al L(-1), (v) manure with alum at 430 mg Al L(-1), and (vi) manure with aluminum chloride at 430 mg Al L(-1). Manure application rates were equivalent to approximately 125 kg N ha(-1). Alum and aluminum chloride additions lowered soluble reactive phosphorus (SRP) levels from about 130 mg P L(-1) to approximately 30 mg P L(-1) at low rates. At high rates, SRP levels in swine manure were around 1 mg P L(-1). Soluble reactive P concentrations in runoff were 5.50, 3.66, 3.00, 0.87, 0.87, and 0.55 mg P L(-1), for normal manure, low alum, low aluminum chloride, high alum, high aluminum chloride, and unfertilized control plots, respectively. Hence, high alum and aluminum chloride reduced SRP concentrations in runoff by 84% and were not statistically different from SRP concentrations in runoff from unfertilized control plots. These data indicate that treating swine manure with alum or aluminum chloride could result in significant reductions in nonpoint-source P runoff. PMID- 11401291 TI - Tillage and nutrient source effects on surface and subsurface water quality at corn planting. AB - This study quantified the effects of tillage (moldboard plowing [MP], ridge tillage [RT]) and nutrient source (manure and commercial fertilizer [urea and triple superphosphate]) on sediment, NH4+ -N, NO3- -N, total P, particulate P, and soluble P losses in surface runoff and subsurface tile drainage from a clay loam soil. Treatment effects were evaluated using simulated rainfall immediately after corn (Zea mays L.) planting, the most vulnerable period for soil erosion and water quality degradation. Sediment, total P, soluble P, and NH4+ -N losses mainly occurred in surface runoff. The NO3- -N losses primarily occurred in subsurface tile drainage. In combined (surface and subsurface) flow, the MP treatment resulted in nearly two times greater sediment loss than RT (P < 0.01). Ridge tillage with urea lost at least 11 times more NH4+ -N than any other treatment (P < 0.01). Ridge tillage with manure also had the most total and soluble P losses of all treatments (P < 0.01). If all water quality parameters were equally important, then moldboard plow with manure would result in least water quality degradation of the combined flow followed by moldboard plow with urea or ridge tillage with urea (equivalent losses) and ridge tillage with manure. Tillage systems that do not incorporate surface residue and amendments appear to be more vulnerable to soluble nutrient losses mainly in surface runoff but also in subsurface drainage (due to macropore flow). Tillage systems that thoroughly mix residue and amendments in surface soil appear to be more prone to sediment and sediment-associated nutrient (particulate P) losses via surface runoff. PMID- 11401292 TI - Seeing the wood through the trees: a review of techniques for distinguishing green fluorescent protein from endogenous autofluorescence. PMID- 11401293 TI - HIV-1 reverse transcriptase interaction with model RNA-DNA duplexes. AB - HIV-1 reverse transcriptase (HIV-1 RT) is a multifunctional enzyme responsible for converting viral RNA into preintegrative DNA during the early stages of viral infection. DNA polymerase and RNase H activities are required, and several conformationally distinct primer-templates must be accommodated by the enzyme during the process. Parameters of interaction between model substrates (ligands) and HIV-1 RT (wild type p66/p51 and the RNase H-deficient mutant p66(E478Q)/p51) (analytes) were estimated by surface plasmon resonance at 25 degrees C, pH 8.0. Binding of RT to the ligands is specific and can be analyzed using a conventional 1:1 binding algorithm. RNA-DNA hybrids with 5'-template overhangs of 6 and 12 nucleotides bind to RT approximately one order of magnitude stronger than the corresponding 36-mer with blunt ends due to slower dissociation. Immobilization of the latter through either the 5'-end of RNA or DNA strand does not change the equilibrium constant (K(D)) for wild-type RT but the values of kinetic constants of association and dissociation differ significantly. For the p66(E478Q)/p51 enzyme, orientation effects are notable even altering the K(D) value. Binding of the p66(E478Q)/p51 to any RNA-DNA hybrids is slightly stronger compared with wild type. Data can be interpreted in terms of the mechanism of reverse transcription. PMID- 11401294 TI - Determination of interaction kinetic constants for HIV-1 protease inhibitors using optical biosensor technology. AB - The interaction between HIV-1 protease and inhibitors has been studied with optical biosensor technology. Optimized experimental procedures and mathematical analysis permitted determination of association and dissociation rate constants. A sensor surface with native enzyme was unstable and exhibited a drift that was influenced by the binding of inhibitor. This was hypothesized to be due to a specific mechanism involving autoproteolysis and/or dimer dissociation. The use of a mutant predicted to be less susceptible to autoproteolysis (Q7K) than wild type enzyme resulted in a minor effect on surface stability, while a completely stable surface was obtained by treatment of the immobilized enzyme with N-ethyl N'-(dimethylaminopropyl)-carbodiimide and N-hydroxysuccinimide; the most stable surface was achieved by chemically modifying the Q7K enzyme. The stabilized surface was enzymatically active and the interaction with inhibitors was similar to that for native enzyme. Several of the inhibitors had very high association rates, and estimation of kinetic constants was therefore performed with a binding equation accounting for limited mass transport. Of the clinical inhibitors studied, saquinavir had the highest affinity for the enzyme, a result of the lowest dissociation rate. Although the dissociation rate for ritonavir was sixfold faster, the affinity was only twofold lower than that for saquinavir since the association rate was threefold faster. Nelfinavir and indinavir exhibited lower affinities relative to the other inhibitors, a consequence of a slower association for nelfinavir and a relatively fast dissociation for indinavir. These results show that biosensor-based interaction studies can resolve affinity into association and dissociation rates, and that these are characteristic parameters for the interaction between enzymes and inhibitors. PMID- 11401295 TI - A duplexed microsphere-based fluorescent immunoassay. AB - Microsphere-based immunoassays are described for the simultaneous measurement of the clinically important drugs digoxin and theophylline. Competitive immunoassays were performed using haptenized microspheres and antibodies labeled with horseradish peroxidase. Enzyme-catalyzed reporter deposition (CARD) resulted in immunofluorescence signal amplification. Two encoding dyes were used to differentiate analytical signals from microspheres containing assays for the two analytes. An epifluorescence microscope and a CCD camera interfaced with a computer were utilized to measure fluorescence signals of individual microspheres. The microspheres from a duplexed assay were mounted on microscope slides as well as inserted into wells etched into the distal ends of optical imaging fibers. Fluorescence images from both formats were captured. In the experiments using microscope slides, the immunoassays were successfully duplexed and only marginal interferences at high analyte concentrations were observed. Preliminary results suggest that simultaneous determination of the two analytes using a fiber-based sensor-array format is feasible, but requires further development before precise quantitative analyses are possible. PMID- 11401296 TI - Pulse electrophoresis of muscle myosin heavy chains in sodium dodecyl sulfate polyacrylamide gels. AB - We have developed a new method that provides enhanced resolution of myosin heavy chain (MHC) isoforms by sodium dodecyl sulfate--polyacrylamide gel electrophoresis (SDS-PAGE). The key feature of this protocol involves the application of current to slab SDS gels in a pulsatile, repetitive manner rather than continuously as in standard gel systems. This protocol, designated pulse electrophoresis, was achieved by means of a device that intermittently gates the output of a conventional power supply. When used in long (32 cm) separating gels, pulse electrophoresis not only significantly improves the resolution of MHC isoforms compared to conventional systems, but also reduces common artifacts associated with long running times, such as blurred bands and comingling of closely spaced bands. In addition to the increased resolution of protein bands, pulse electrophoresis also allows detection of bands corresponding to previously unidentified MHC isoforms in mammalian and avian tissue. In rat myocardium, for example, pulse electrophoresis revealed three MHC isoform bands, two of which appeared to correspond to two alpha-MHC subspecies. Alternative splicing of the rat alpha-MHC gene is known to generate two isoform species differing by inclusion (or exclusion) of a single glutamine residue, whose relative levels of expression correspond nicely with the amounts of each band identified in this study. Therefore, we cannot rule out that the system presented here may be sufficiently sensitive to differentiate between high molecular weight proteins differing in a single amino acid. PMID- 11401297 TI - Determination of the relative expression levels of rubisco small subunit genes in Arabidopsis by rapid amplification of cDNA ends. AB - Multigene families are common in higher organisms. However, due to the close similarities between members, it is often difficult to assess the individual contribution of each gene to the overall expression of the family. In Arabidopsis thaliana, there are four genes encoding the small subunits (SSU) of ribulose-1.5 bisphosphate carboxylase oxygenase (rubisco) whose nucleotide sequences are up to 98.4% identical. In order to overcome the technical limitations associated with gene-specific probes (or primers) commonly used in existing methods, we developed a new gene expression assay based on the RACE (rapid amplification of cDNA ends) technique with a single pair of primers. With this RACE gene expression assay, we were able to determine the relative transcript levels between four Arabidopsis SSU genes. We found that the relative SSU gene expression differed significantly between plants grown at different temperatures. Our observation raises the possibility that an adaptation of rubisco to the environment may be achieved through the specific synthesis of the SSU proteins, which is determined by the relative expression levels between the SSU genes. PMID- 11401298 TI - Directly coupled high-performance liquid chromatography and nuclear magnetic resonance spectroscopic with chemometric studies on metabolic variation in Sprague--Dawley rats. AB - We report here the first combined use of NMR-PR (pattern recognition) analysis and directly coupled HPLC--NMR analysis to identify metabolic subpopulations in normal laboratory animals and their discriminating endogenous urinary biomarkers. Urine samples obtained from control Sprague-Dawley rats (n = 68) were analyzed using (1)H NMR spectroscopy and principal components (PC) analysis to investigate physiological variability. Two distinct subpopulations of animals were classified based on metabolite excretion profiles. Analysis of the PC loadings established the spectral regions that were responsible for classification of the subpopulations and was used to direct the identification of biomarkers using a directly coupled HPLC--NMR analysis. One population had low urinary hippurate levels together with an increased concentration of 3-(3-hydroxyphenyl)propionic acid (3-HPPA)and 3-hydroxycinnamic acid (3-HCA). The other subpopulation excreted high levels of hippurate. Thus, we report the bimodal occurrence of hippuric acid and chlorogenic acid metabolites in a genetically homogeneous population of rats maintained under identical conditions, which may have significance in relation to the understanding of the consequences of biochemical variation in animals used for drug toxicity testing. PMID- 11401299 TI - High-throughput fluorogenic assay for determination of botulinum type B neurotoxin protease activity. AB - Botulinum neurotoxins are responsible for botulism, a flaccid muscular paralysis caused by inhibition of acetylcholine release at the neuromuscular junction. This occurs by cleavage of conserved proteins involved in exocytosis such as synaptobrevin by the zinc metallopeptidase activity of the light chain of some botulinum neurotoxins. Botulism, for which there is presently no therapy available, is a relatively widespread disease that may result in death. Consequently, the development of drugs able to inhibit the hydrolytic activity of these neurotoxins is of great interest. Design and screening of such inhibitors could be largely facilitated by using high-throughput assays. With this aim, a novel in vitro test for quantifying the proteolytic activity of botulinum type B neurotoxin was developed. The substrate is the 60--94 fragment of human synaptobrevin-1 which was modified by introduction of the fluorescent amino acid l-pyrenylalanine in position 74 and a p-nitrophenylalanyl residue as quenching group in position 77. The cleavage of Syb 60-94 [Pya(74), Nop(77)] by the toxin active chain occurs selectively between residues 76 and 77 as in the case of the unmodified synaptobrevin and is directly quantified by measuring the strong fluorescence of the formed metabolite Syb 60-76 [Pya(74)]. This is the easiest, quickest, and cheapest assay described to date for measuring the proteolytic activity of botulinum type B neurotoxin. It can be easily automated for high throughput screening. Moreover, amounts of about 3.5 pg/ml of botulinum type B neurotoxin could be detected by this method. PMID- 11401300 TI - Analysis of recombinant adenoviruses using an integrated microfluidic chip-based system. AB - The use of recombinant virus for gene therapy requires rigorous quality control methods to ensure that the viral vector preparations are functional and safe. A viral identity test is performed in which the viral payload, or transgene, is PCR amplified, followed by digestion with restriction enzymes that yields a characteristic "fingerprint." These DNA fragments are typically analyzed by agarose gel electrophoresis. The ethidium bromide-stained gels are photographed or scanned and the results are sufficient for a qualitative or semiquantitative identity confirmation of the viral product. We have investigated the use of an integrated microfluidic chip-based system as a new tool in the quality control testing of a recombinant, adenoviral, gene therapy product. The chip-based method was found to be very sensitive, requiring 100-fold less sample and only one-third the time compared to the agarose gel method. The automated data analysis sizes and quantitates the DNA fragments, thus yielding a more thorough, reproducible, sensitive, and rapid analysis. PMID- 11401301 TI - Development of a robust scintillation proximity assay for protein tyrosine phosphatase 1B using the catalytically inactive (C215S) mutant. AB - Protein tyrosine phosphatases are a class of enzymes that function to modulate tyrosine phosphorylation of cellular proteins and play an essential role in regulating cell function. PTP1B has been implicated in the negative regulation of the insulin signaling pathway by dephosphorylating the activated insulin receptor. Inhibiting this phosphatase and preventing the insulin-receptor downregulation has been suggested as a target for the treatment of Type II diabetes. A high-throughput screen for inhibitors of PTP1B was developed using a scintillation proximity assay (SPA) with GST-- or FLAG--PTP1B((1-320)) and a potent [(3)H]-tripeptide inhibitor. The problem of interference from extraneous oxidizing and alkylating agents which react with the cysteine active-site nucleophile was overcome by the use of the catalytically inactive C215S form of the native enzyme (GST--PTP1B(C215S)). The GST--PTP1B was linked to the protein A scintillation bead via GST antibody. The radiolabeled inhibitor when bound to the enzyme gave a radioactive signal that was competed away by the unknown competitive compounds. Further utility of PTP1B(C215S) was demonstrated by mixing in the same well both the catalytically inactive GST--PTP1B(C215S) and the catalytically active FLAG--CD45 with an inhibitor. Both a binding and kinetic assay was then performed in the same 96-well plate with the inhibition results determined for the PTP1B(C215S) (binding assay) and CD45 (activity assay). In this way inhibitors could be differentiated between the two phosphatases under identical assay conditions in one 96-well assay plate. The use of a mutant to reduce interference in a binding assay and compare with activity assays is also amenable for most cysteine active-site proteases. PMID- 11401302 TI - High-performance liquid chromatography-based determination of total isothiocyanate levels in human plasma: application to studies with 2-phenethyl isothiocyanate. AB - Dietary and pharmacologic isothiocyanates (ITCs) may play a role in reducing the risk of certain cancers. The quantification of ITCs in humans is important both for epidemiological and pharmacokinetic studies. We describe a modification of an HPLC-based assay of urinary ITCs for use with human plasma. The assay utilizes the cyclocondensation reaction of 1,2-benzenedithiol with ITCs present in human plasma, followed by a two-step hexane extraction and analysis by HPLC using UV detection at 365 nm. The method shows linearity and reproducibility with human plasma over a range of 49-3003 nM phenethyl isothiocyanate (PEITC) (r(2) = 0.996 +/- 0.003). A similar degree of linearity was seen with two other biologically occurring conjugates of PEITC: PEITC--N-acetylcysteine (PEITC--NAC) and PEITC- glutathione (PEITC--GSH). The recovery of PEITC assessed on multiple days was 96.6 +/- 1.5% and was 100% for PEITC--GSH and PEITC--NAC. The reproducibility of the assay on multiday samplings showed a mean %CV of 6.5 +/- 0.3% for PEITC, 6.4 +/- 4.3 for PEITC--NAC and 12.3 +/- 3.9 for PEITC--GSH. In clinical studies, mean plasma ITC level of 413 +/- 193 nM PEITC equivalents was determined for a non dietary-controlled group of 23 subjects. Multiday analysis data from pharmacokinetic plasma sets of 3 subjects taking a single dose of PEITC at 40 mg showed a good CV (range: 16-21%). The applicability of the methodology to pharmacokinetic studies of PEITC in humans is demonstrated. PMID- 11401303 TI - Selective and sensitive detection of pectin lyase activity using a colorimetric test: application to the screening of microorganisms possessing pectin lyase activity. AB - Several methods have been described for the detection and quantification of polygalacturonase (PG) and pectin lyase (PL) activities. The most frequently used tests are the Nelson method using copper(II) and an arsenomolybdate reagent to detect PG activity, and the colorimetric method using thiobarbituric acid (TBA) to detect PL activity. We observed that none of these methods are suitable to differentiate between these two enzymatic activities. Therefore, we optimized the test conditions of the TBA method. As a result, the detection of the enzymatic beta-elimination (PL activity) became sensitive and selective. A basic pretreatment at 80 degrees C for 5 min of the solution which contains the pectin fragments of the PL activity furnished aldehydes which were condensed with TBA or its derivatives. After acidification of the medium, a pink fluorescent dye was detected spectrophotochemically (lambda = 550 nm). The interference of galacturonic acid or oligomers resulting from PG activity was completely eliminated. The most sensitive reagent was N-(pyridin-2-yl)-thiobarbituric acid. The application of this method with the new reagent was extended to the screening of microorganisms possessing the PL activity. The obtained results confirm that Aspergillus niger strain and a Saccharomyces cerevisiae SCPP strain possess this activity. PMID- 11401304 TI - An automated high-performance liquid chromatography procedure for the quantitation of L- and D-amino acids by means of stepwise precolumn derivatization. PMID- 11401305 TI - Modified silver staining for RNA and DNA in agarose gels. PMID- 11401306 TI - Polyacrylamide gel electrophoresis without a stacking gel: use of amino acids as electrolytes. PMID- 11401307 TI - A facile synthesis of homocysteine-cysteine mixed disulfide. PMID- 11401308 TI - Patients' preferences for adjuvant chemotherapy in early-stage breast cancer: is treatment worthwhile? AB - When making decisions about adjuvant chemotherapy for early-stage breast cancer, costs and benefits of treatment should be carefully weighed. In this process, patients' preferences are of major importance. The objectives of the present study were: (1) to determine the minimum benefits that patients need to find chemotherapy acceptable, and (2) to explore potential preference determinants, namely: positive experience of the treatment, reconciliation with the treatment decision, and demographic variables. Preferences were elicited from patients scheduled for adjuvant chemotherapy (chemotherapy group: n = 38) before (T(1)), during (T(2)), and 1 month after chemotherapy (T(3)), and were compared to responses from patients not scheduled for chemotherapy (no-chemotherapy group: n = 38). The patients were asked, for a hypothetical situation, to indicate the minimum benefit (in terms of improved 5-year disease-free survival) to find adjuvant chemotherapy acceptable. In the chemotherapy group, the median benefit was 1% at all 3 measurement points. In the no-chemotherapy group the attitude towards chemotherapy became more negative over time, although not statistically significantly so (T(1): 12%, T(2): 15%, T(3): 15%; P = 0.10). At all measurement points, the patients in the chemotherapy group indicated that they would accept chemotherapy for significantly (P< 0.01) less benefit than the patients in the no chemotherapy group. Of the demographic variables, age was related to preferences, but only at T(2)and only in the no-chemotherapy group. The more positive attitude towards chemotherapy and the stability of preferences in the chemotherapy group indicated that reconciliation with the treatment decision was a more important determinant of patients' preferences than positive experience of the treatment. PMID- 11401309 TI - Pamidronate improves the quality of life and induces clinical remission of bone metastases in patients with thyroid cancer. AB - Skeletal metastases from thyroid cancer are poorly responsive to medical or radioiodine treatment. Bone destruction in skeletal metastases results from osteoclast-induced bone resorption. Therefore, a new approach in the therapy of bone metastases consists in using aminobisphosphonates, such as pamidronate, which are potent inhibitors of osteoclastic activity. In the present study, 10 thyroid cancer patients with painful osteolytic bone metastases were administered pamidronate (90 mg, as a 2 hour intravenous infusion) monthly for 12 consecutive cycles. Bone pain, quality of life, performance status, analgesic consumption and disease staging were evaluated before and during the trial. The patients who had been administered pamidronate showed a significant decrease in bone pain (P = 0.0052). Performance status improved nearly significantly (P = 0.051), while the quality of life showed a remarkable amelioration. However, no significant decrease in analgesic consumption was recorded. Partial radiographic response of bone lesions was observed in 2/10 patients. The side effects of pamidronate were mild and transient. In conclusion, monthly infusion of pamidronate is a well tolerated treatment that induces significant relief from bone pain and improves the quality of life of thyroid cancer patients with symptomatic and osteolytic bone metastases. PMID- 11401310 TI - High-dose paclitaxel in combination with doxorubicin, cyclophosphamide and peripheral blood progenitor cell rescue in patients with high-risk primary and responding metastatic breast carcinoma: toxicity profile, relationship to paclitaxel pharmacokinetics and short-term outcome. AB - We assessed the feasibility and pharmacokinetics of high-dose infusional paclitaxel in combination with doxorubicin, cyclophosphamide, and peripheral blood progenitor cell rescue. Between October 1995 and June 1998, 63 patients with high-risk primary [stage II with >or= 10 axillary nodes involved, stage IIIA or stage IIIB inflammatory carcinoma (n = 53)] or with stage IV responsive breast cancer (n = 10) received paclitaxel 150-775 mg/m(2)infused over 24 hours, doxorubicin 165 mg/m(2)as a continuous infusion over 96 hours, and cyclophosphamide 100 mg kg(-1). There were no treatment-related deaths. Dose limiting toxicity was reversible, predominantly sensory neuropathy following administration of paclitaxel at the 775 mg/m(2) dose level. Paclitaxel pharmacokinetics were non-linear at higher dose levels; higher paclitaxel dose level, AUC, and peak concentrations were associated with increased incidence of paraesthesias. No correlation between stomatitis, haematopoietic toxicities, and paclitaxel dose or pharmacokinetics was found. Kaplan-Meier estimates of 30-month event-free and overall survival for patients with primary breast carcinoma are 65% (95% CI; 51-83%) and 77% (95% CI; 64-93%). Paclitaxel up to 725 mg/m(2) infused over 24 hours in combination with with doxorubicin 165 mg/m(2) and cyclophosphamide 100 mg kg(-1) is tolerable. A randomized study testing this regimen against high-dose carboplatin, thiotepa and cyclophosphamide (STAMP V) is currently ongoing. PMID- 11401311 TI - Proteolysis-inducing factor is expressed in tumours of patients with gastrointestinal cancers and correlates with weight loss. AB - Proteolysis-inducing factor (PIF), a novel cachectic factor, is detectable in the urine of cancer patients experiencing weight loss. We report the expression of PIF in gastrointestinal cancers, and a correlation between PIF expression in tumours, its detection in urine and weight-loss. These data provide the first direct evidence that tumours are the source of PIF in humans. PMID- 11401312 TI - Pathological serosa and node-based classification accurately predicts gastric cancer recurrence risk and outcome, and determines potential and limitation of a Japanese-style extensive surgery for Western patients: a prospective with quality control 10-year follow-up study. AB - UICC classification accurately predicts overall survival but not recurrence-risk. We report here data of overall and first site-specific recurrence following curative surgery useful for the development of recurrence-oriented preventive target therapies. Patients who underwent resection for gastric cancer were stratified according to curability of surgery [curative (R0) vs non-curative resection], extent of surgery [limited (D1) vs extended (D2) node dissection] and pathological nodal/serosal status. The intent-to-treat principle, log-rank test and Cox regression analysis were used for statistical analysis of time-to-event (recurrence, death) endpoints. Curative resection only produced a chance of cure whereas survival was very poor following non-curative resection (P < 0.0001). For D2 R0 subgroup of patients, a pathological serosa and a node state-based classification into three groups, proved to be of clinical implication. Risk of recurrence after a median follow-up of 92 months was low among patients with both serosa and node-negative cancer (first group; 11%), moderate among those with either serosa or node-positive cancer (second group; 53%) and very high among those with both serosa and node-positive cancer (third group; 83%). In multivariate analysis, the relative risks of recurrence and death from gastric cancer among patients in the second and third groups, as compared to those in the first, were 7.07 (95% CI, 2.36-21.17; P = 0.0002) and 16.19 (95% CI, 5.76-45.54; P < 0.0001) respectively. First site-specific recurrence analysis revealed: low rate of loco-regional recurrence alone (12%), serosa state determinant factor of the site-recurrence (peritoneal for serosa-positive and haematogenous for serosa negative cancers) and dramatic increase of all types of recurrence by the presence of nodal metastases. Our findings demonstrate that a pathological serosa and node-based classification is very simple and predicts accurately site specific recurrence-risks. Furthermore they reveal that risk of recurrence following curative D2 surgery alone is low for serosa- and node-negative cancers, but very high in serosa- and node-positive cancers suggesting the need for new therapeutic strategies in this subgroup of patients. PMID- 11401313 TI - Concentration of vascular endothelial growth factor in the tumour tissue as a prognostic factor of soft tissue sarcomas. AB - Previous studies have shown that the prognosis of patients who have tumours with high microvessel density (MVD) is worse than that of patients who have a lower density in a variety of cancers. In this study, we investigated the clinical relevance of neovascularity assessed by MVD and the concentration of vascular endothelial growth factor (VEGF) in the tumour tissue of patients with soft tissue sarcoma in comparison with major clinicohistologic parameters by univariate and multivariate analysis. In 115 patients with soft tissue sarcoma, MVD was measured by counting vessels stained with factor VIII antibody. The concentration of VEGF in the tumour tissue was determined by enzyme-linked immunosorbent assay. These parameters were then compared with disease outcome. The concentration of VEGF in the tumour tissue, but not MVD, was found to be correlated with disease outcome in patients with soft tissue, sarcoma. VEGF concentration in the tumour tissue showed a relationship with the clinical stage and histologic grade of the tumour. There was no significant difference in the levels of tissue VEGF concentration and MVD among soft tissue sarcomas classified according to histologic type. The level of tissue VEGF concentration in patients who had subsequent local recurrence and metastasis were significantly higher than the respective values in patients who did not have such disease outcome. No significant correlation existed between MVD and the concentration of VEGF in the tumour tissue. Univariate analysis showed that a high tissue VEGF concentration was associated with poor overall survival of the patient and a greater probability that local recurrence and metastasis had occurred. Multivariate analysis revealed that the tissue concentration of VEGF is an independent prognostic factor for the disease outcome of patients with soft tissue sarcoma. VEGF concentration in the tumour tissue, but not MVD, is an additional prognostic parameter for disease outcome in patients with soft tissue sarcoma, regardless of histologic type. PMID- 11401314 TI - Human papillomavirus testing in primary screening for the detection of high-grade cervical lesions: a study of 7932 women. AB - High-risk human papillomaviruses (HR-HPV) are the necessary cause of cervical carcinomas. To determine whether HPR-HPV DNA detection in primary routine screening could represent a sensitive and reliable technique for the detection of high-grade squamous intraepithelial lesions (HGSIL), laboratory analysis using 2 cytologic techniques (conventional and liquid-based), HPV testing with Hybrid Capture II assay (HC-II), followed by colposcopic examination of women with abnormal cervical finding and/or persistent HR-HPV infection, was conducted in 7932 women who had routine cervical examination. The sensitivity of HPV testing for detecting a histologically proven HGSIL was 100%, higher than that of conventional (68.1%) and liquid-based (87.8%) cytology. The low specificities of 85.6% and 87.3% of HPV testing slightly increased to 88.4% and 90.1% if HPV testing was reserved for woman >30 years old. The quantitative approach provided by the HC-II assay for the assessment of the viral load was not reliable for predicting HGSIL in normal smears. HR-HPV testing could be proposed in primary screening in association with cytology. With conventional cytology it significantly improves the detection of HGSIL. With the use of the same cervical scrape for HPV testing and liquid-based cytology, HR-HPV testing would allow to select positive samples treated in a second time for cytology which gives a good specificity. PMID- 11401315 TI - Modulation of TcR/CD3-zeta chain expression by a circulating factor derived from ovarian cancer patients. AB - In women with ovarian cancer, suppression of components of the immune system may promote tumour development. Previous studies in ovarian cancer have demonstrated that decreased expression and function of the T-cell receptor (TcR)-associated signal transducing zeta-chain correlates with deficient immune responsiveness of T cells. In this study, sera and ascitic fluids obtained from woman with advanced ovarian cancer were found to suppress the expression of TcR-associated zeta chain. This suppression of zeta chain expression was dose-dependent and was not observed with biologic fluids obtained from healthy women. The factor responsible for the loss of zeta chain was purified from ascites and characterized as a protein with an appropriate molecular weight of 14 kD. Suppression of T-cell TcR zeta was specific, since neither lck nor ZAP-70 expression was affected, while zeta chain was almost completely suppressed. This selective suppression of TcR zeta expression by the 14 kD ascites-derived factor was shown to operate at the mRNA level. By defining the mechanism through which this protein modulates TcR zeta chain levels, it might be possible to ultimately prevent the suppressive influences of the tumour microenvironment and restor immune competence in patients with ovarian carcinoma. PMID- 11401316 TI - Detection of functional PTEN lipid phosphatase protein and enzyme activity in squamous cell carcinomas of the head and neck, despite loss of heterozygosity at this locus. AB - The human tumour suppressor gene PTEN located at 10q23 is mutated in a variety of tumour types particularly metastatic cases and in the germline of some individuals with Cowdens cancer predisposition syndrome. We have assessed the status of PTEN and associated pathways in cell lines derived from 19 squamous cell carcinomas of the head and neck. Loss of heterozygosity is evident at, or close to the PTEN gene in 5 cases, however there were no mutations in the remaining alleles. Furthermore by Western analysis PTEN protein levels are normal in all of these SCC-HN tumours and cell lines. To assess the possibility that PTEN may be inactivated by another mechanism, we characterized lipid phosphatase levels and from a specific PIP3 biochemical assay it is clear that PTEN is functionally active in all 19 human SCCs. Our data strongly suggest the possibility that a tumour suppressor gene associated with development of SCC-HN, other than PTEN, is located in this chromosomal region. This gene does not appear to be MXI-1, which has been implicated in some other human tumour types. PTEN is an important negative regulator of PI3Kinase, of which subunit alpha is frequently amplified in SCC-HN. To examine the possibility that PI3K is upregulated by amplification in this tumour set we assessed the phosphorylation status of Akt, a downstream target of PI3K. In all cases there is no detectable increase in Akt phosphorylation. Therefore there is no detectable defect in the PI3K pathway in SCC-HN suggesting that the reason for 3q26.3 over-representation may be due to genes other than PI3K110alpha. PMID- 11401317 TI - p53 missense but not truncation mutations are associated with low levels of p21(CIP1/WAF1) mRNA expression in primary human sarcomas. AB - Many growth-suppressing signals converge to control the levels of the CDK inhibitor p21(CIP1/WAF1). Some human cancers exhibit low levels of expression of p21(CIP1/WAF1) and mutations in p53 have been implicated in this down-regulation. To evaluate whether the presence of p53 mutations was related to the in vivo expression of p21(CIP1/WAF1) mRNA in sarcomas we measured the p21(CIP1/WAF1) mRNA levels for a group of 71 primary bone and soft tissue tumours with known p53 status. As expected, most tumours with p53 mutations expressed low levels of p21(CIP1/WAF1)mRNA. However, we identified a group of tumours with p53 gene mutations that exhibited normal or higher levels of p21(CIP1/WAF1) mRNA. The p53 mutations in the latter group were not the common missense mutations in exons 4 9, but were predominantly nonsense mutations predicted to result in truncation of the p53 protein. The results of this study suggest that different types of p53 mutations can have different effects on the expression of downstream genes such as p21(CIP1/WAF1) in human sarcomas. PMID- 11401318 TI - Peroxisome proliferator-activated receptor gamma ligand-induced growth inhibition of human hepatocellular carcinoma. AB - Peroxisome proliferator-activated receptor gamma (PPARgamma) ligands have been implicated in the growth inhibition and differentiation of certain human cancers with diverse tissue origin. In this study, expression of PPARgamma in human hepatocellular carcinoma (HCC) and the effect of PPARgamma ligands on HCC cells were investigated in vitro using Hep G2, HuH-7, KYN-1 and KYN-2 cell lines. All cell lines were found to express functionally active PPARgamma and a marked growth inhibition was induced by thiazolidinedione ligands troglitazone, and pioglitazone as well as with its natural ligand 15-deoxy-Delta(12,14) prostaglandin J(2). The growth inhibitory effect was associated with a dose dependent inhibition of DNA synthesis, cell cycle progression and alpha fetoprotein expression. PMID- 11401319 TI - Effect of a tumour-produced lipid-mobilizing factor on protein synthesis and degradation. AB - Treatment of murine myoblasts, myotubes and tumour cells with a tumour-produced lipid mobilizing factor (LMF), caused a concentration-dependent stimulation of protein synthesis, within a 24 h period. There was no effect on cell number or [(3)H] thymidine incorporation, but a similar concentration-dependent stimulation of 2-deoxyglucose uptake. LMF produced an increase in intracellular cyclic AMP levels, which was linearly (r(2)= 0.973) related to the increase in protein synthesis. The effect of LMF was attenuated by the adenylate cyclase inhibitor MDL(12330A), and was additive with the stimulation produced by forskolin. Both propranolol (10 microM) and the specific beta(3)-adrenergic receptor antagonist SR 59230A (10(-5)M), significantly reduced the stimulation of protein synthesis induced by LMF. Protein synthesis was also increased by 69% (P = 0.006) in soleus muscles of mice administered LMF, while there was a 26% decrease in protein degradation (P = 0.03). While LMF had no effect on the lysosomal enzymes, cathepsins B and L, there was a decrease in proteasome activity, as determined both by the 'chymotrypsin-like' enzyme activity, as well as expression of proteasome alpha-type subunits, determined by Western blotting. These results show that in addition to its lipid-mobilizing activity LMF also increases protein accumulation in skeletal muscle both by an increase in protein synthesis and a decrease in protein catabolism. PMID- 11401320 TI - FGF-1 and FGF-2 modulate the E-cadherin/catenin system in pancreatic adenocarcinoma cell lines. AB - Fibroblast growth factors (FGFs) and fibroblast growth factor receptors (FGFRs) have been increasingly recognized to play an important role in the pathobiology of pancreatic malignancy. We have investigated the effects of FGF-1 and FGF-2 on the behaviour and adhesion properties of human pancreatic adenocarcinoma cell lines (BxPc3, T3M4 and HPAF) that were previously characterised for the expression of FGFRs. Here we show that exposure to FGF-1 and FGF-2 leads to significant and dose-dependent increase in E-cadherin-dependent cell-cell adhesion, tubular differentiation, and a reduced capacity to invade collagen gels. FGF stimulation produces phosphorylation of E-cadherin and beta-catenin on tyrosine residues, as well as increased E-cadherin localisation to the cytoplasmic membrane and association with FGFR1 demonstrable by coimmunoprecipitation. These results demonstrate that FGF-1 and FGF-2 may be involved in the regulation of cell adhesion, differentiation and invasion of pancreatic cancer. PMID- 11401321 TI - Spectrum of matrix metalloproteinase expression in primary and metastatic colon cancer: relationship to the tissue inhibitors of metalloproteinases and membrane type-1-matrix metalloproteinase. AB - The matrix metalloproteinases, MMP-2 and MMP-9, are capable of degrading components of the basement membrane, a vital barrier breached during the progression of colorectal cancer. The regulation of MMP-2 activation and subsequent targets is vital to understanding the metastatic process. MMP-2 was not expressed by colorectal cancer cells (C170 and C170HM(2)) in vitro but by stromal fibroblasts (46BR.1GI). There was induction of this MMP upon transwell co cultivation of the colon cancer cells with the fibroblasts but in vivo growth did not lead to a similar increase in the metastatic tumour cells (C170HM(2)), MMP-2 again being attributed to the stromal cells. MMP-2 mRNA was overexpressed in human colorectal tumours compared to normal colorectal tissue, which correlated with Dukes' stage and immunolocalized to the stromal compartment of the tumour tissue. The active form of the MMP-2 enzyme was also present in the colorectal tumour tissue (7/8) but essentially absent in all normal colon samples examined (1/8). MMP-2 activation was not related to an increase in MT-1-MMP mRNA or a decrease in the specific inhibitor TIMP-2 in human tissue. There was however an increase in MMP-2/TIMP-2 ratio in tumour compared to normal. MMP-9, a target of active MMP-2, was present in the metastatic cell line but expression was down regulated in the tumour cells in vivo, gelatin analysis revealed that MMP-9 was almost entirely attributable to the murine host, confirmed by PCR. There was no increase in mRNA for MMP-9 or its specific inhibitor TIMP-1 in colorectal tumour tissue compared to normal, MMP-9 protein localized to the inflammatory infiltrate. Fibroblast cells may provide malignant epithelial cells with a ready source of enzyme which is crucial to the metastatic process. PMID- 11401322 TI - Measurement of the critical DNA lesions produced by antibody-directed enzyme prodrug therapy (ADEPT) in vitro, in vivo and in clinical material. AB - An antibody-directed enzyme prodrug therapy (ADEPT) system against CEA-positive tumours is currently in phase I clinical trials. It consists of a prodrug, 4-[N,N bis(2-iodoethyl) amino] phenoxycarbonyl L -glutamic acid (ZD2767P) and a conjugate of the F(ab')(2) anti-CEA antibody A5B7 and the bacterial enzyme carboxypeptidase G2 (CPG2). ZD2767P is converted by antibody-targeted CPG2 into an active bifunctional alkylating drug (ZD2767) at the tumour site. The IC(50) value of the prodrug against the human colorectal tumour LS174T cell line was 55 +/- 9 microM following a 1 h exposure. In contrast, co-incubation of ZD2767P with CPG2 resulted in 229-fold increase in activity. Using a modified comet assay, DNA interstrand cross links (ISC) were detected within 1 h of ZD2767P + CPG2 treatment and were repaired by 24 h. A clear dose-response was seen between the level of ISC, growth inhibition and ZD2767 concentration. Administration of a therapeutic dose of ZD2767P 72 h after the F(ab')(2) A5B7 conjugate to mice bearing LS147T xenografts resulted in extensive ISC in the tumour after 1 h; repair was seen at 24 h. Tumour biopsies and peripheral lymphocytes were studied in 5 patients on the ADEPT phase I clinical trial. In 4 patients no ISC were detected. These patients also demonstrated poor localization of conjugate and no tumour response was seen. However a significant level of ISC was detected in one tumour biopsy, which also showed evidence of conjugate localization and clinical response. These studies demonstrate the application of the comet assay in the measurement of ISC in vitro and in clinical material and confirm that activation of ZD2767P results in the formation of DNA crosslinks. PMID- 11401323 TI - Fluoropyrimidine sensitivity of human MCF-7 breast cancer cells stably transfected with human uridine phosphorylase. AB - The relationship between uridine phosphorylase (UP) expression level in cancer cells and the tumour sensitivity to fluoropyrimidines is unclear. In this study, we found that UP overexpression by gene transfer, and the subsequent efficient metabolic activation of 5-fluorouracil (5-FU) by the ribonucleotide pathway, does not increase the fluoropyrimidine sensitivity of MCF-7 human cancer cells. PMID- 11401324 TI - Tumour enhancement with newly developed Mn-metalloporphyrin (HOP-9P) in magnetic resonance imaging of mice. AB - The purpose of the study is to evaluate the tumour enhancing characteristics and biodistribution of a newly developed metalloporphyrin derivative, HOP-9P (13, 17 bis (1-carboxypropionyl) carbamoylethyl-3, 8-bis (1-phenylpropyloxyethyl) 2,7,12,18-tetra- methyl-porphynato manganese (III)). Seven mice bearing SCC VII tumours were imaged using T1-weighted conventional spin echo magnetic resonance images before and 5 min, 2 h and 24 h after intravenous injection of 0.1 mmol/kg of HOP-9P. For the acquired images, signal intensities of the tumour, muscle and oil-phantom were measured. Then, tumor/oil and tumor/muscle signal intensity ratios were calculated. Nineteen mice were sacrificed before or after the administration of HOP-9P (at 5 min, 2 h and 24 h), and the biodistribution of manganese in the tumour, muscle, liver, blood and kidneys was measured using optical emission spectrometers and was expressed as micrograms of manganese per gram of tissue. The tumour/muscle signal intensity ratio at 24 h (3.18 +/- 0.34) was significantly higher than precontrast ratio (1.77 +/- 0.20) (P < 0.05). The biodistribution assessment of manganese demonstrated that HOP-9P gradually and consistently accumulated in the tumour to reach the highest concentration at 24 h (3.49 +/- 1.22 micro gMn/g). It is concluded that HOP-9P is a potential tumour specific MR contrast agent. PMID- 11401326 TI - Laminin-2/4 from human placenta is a better adhesion agent for primary keratinocytes than laminin-1 from EHS sarcoma. AB - A comparison of the adhesion of human primary keratinocytes to laminin-1 from murine EHS sarcoma and laminin-2/4 from human placenta was carried out using two methods, cell adhesion to substrates covered with the laminin isoforms, and interaction of keratinocytes from suspension with latex beads coated with the proteins. Laminin-2/4 was considerably more potent as a promoter of attachment of primary human keratinocytes than laminin-1 (and fibronectin), with increased attachment of cells correlating well with the number of latex bead binding sites. Only small cells of diameter of less than 20 microm bound more than 5 beads. Staining of keratinocytes with involucrin antibodies confirmed the existence of an inverse relationship between laminin-2/4-coated bead binding and differentiation. PMID- 11401327 TI - Participation of the Ca(2+)-calmodulin-activated Kinase II in the control of metaphase-anaphase transition in human cells. AB - Treatment of HeLa S3 cells growing in suspension, and of endothelial cells ECV 304 growing as a monolayer, with the inhibitor of the Ca(2+)-calmodulin activated Kinase II KN-93, blocks cells at metaphase for 15 min (HeLa cells) and 30 min (ECV cells). Thereafter cells resume mitosis and enter anaphase. The inactive isomer KN-92 does not show such effects. The results show the involvement of the CaM K II system in the regulation of the metaphase-anaphase transition whereby the activation of the Kinase II, in particular by calmodulin appears to be affected, the residual autophosphorylation of the CaM K II system apparently sufficing after 15 to 30 min to release the cells into anaphase. The results are compared with the metaphase-blocking effects of the noble gas xenon, where the xenon-induced block can be overcome by small intracellular increases of Ca(2+), thus indicating the CaM K II system as a possible target for xenon. PMID- 11401328 TI - Altered matrix metalloproteinase expression associated with oncogene-mediated cellular transformation and metastasis formation. AB - Matrix metalloproteinase expression was examined in a series of mammalian cell lines of varying degrees of malignant progression. The expression of MMP-2 and MMP-9 was found to correlate with ras-mediated cellular transformation and as a function of malignant potential. Altered MMP-2 and MMP-9 expression was found to correlate also in other oncogene transformed cell lines and the level of expression of both MMP-2 and MMP-9 correlated with metastatic potential. Increased expression of both MMP-2 and MMP-9 was also found in cells which constitutively over-express MAP kinase kinase suggesting that one of the consequences of the persistent activation of the MAP kinase pathway is elevated expression of MMP-2 and MMP-9. Additionally, this study demonstrated a correlation between the expression of MMP-3 (stromelysin-1) and the level of ras expressed in cells and with the cells' ability to form tumors and with malignant potential. The existence of a novel 80 kDa caseinase activity which correlates with ras expression and the ability of the cell to form tumors was also demonstrated. The growth status of transformed cells was also found to be important in determining the expression of MMP-2 mRNA but not MMP-9 mRNA expression, and this expression was cell-type specific. This study also demonstrates that oncogenes can interact to influence and to determine the nature of the matrix metalloproteinases expressed and that this interaction results in a tumorigenic phenotype and, most importantly, contributes to the metastatic phenotype. Alterations in the expression and the regulation of MMPs, particularly MMP-2 and MMP-9, constitute an integral part of the altered growth regulatory program found within transformed cells and in particular, in transformed cells capable of malignant progression. PMID- 11401329 TI - Tyrosine phosphorylation translocates beta-catenin from cell-->cell interface to the cytoplasm, but does not significantly enhance the LEF-1-dependent transactivating function. AB - beta-catenin plays an essential role in cells, not only as a cadherin-associated complex, but also as a signaling molecule in the nucleus. Tyrosine phosphorylation of beta-catenin has been shown to correlate with tumorigenesis, cell migration, and developmental processes. However, its exact effects on downstream targets in the nucleus are not yet clear. In this study, we used HCT 15 colon carcinoma and NIH 3T3 fibroblasts as models to investigate the effects of a phosphotyrosine phosphatase (PTPase) inhibitor on the localization of beta catenin, the binding affinity to LEF-1 (Lymphoid Enhancer Factor), and on LEF-1 dependent transactivation function. Treatment with a PTPase inhibitor, pervanadate, increased the tyrosine phosphorylation of beta-catenin in a time dependent manner and led to its relocation from cell-cell interfaces to the cytoplasm. This phosphorylation/dephosphorylation of beta-catenin does not require its presence at cell-cell interfaces. However, tyrosine phosphorylation of beta-catenin does not change its binding affinity to LEF-1 nor enhance cyclin D1 transactivation, a nuclear target of beta-catenin/LEF-1. This result suggests that tyrosine phosphorylation of beta-catenin has effects on the binding to cadherins in the cytoplasm but not on its LEF-1-dependent transactivating function in the nucleus. PMID- 11401330 TI - Effect of bone morphogenetic protein-6 on haemopoietic stem cells and cytokine production in normal human bone marrow stroma. AB - Normal human bone marrow stroma cells include stem cells for both haemopoietic and osteochondrogenic lineages and express both bone morphogenetic protein (BMP) type I and type II receptors. As a member of the TGF-beta super-family, BMP-6 binds to both BMP type I and type II receptors and is involved in the developmental processes of renal and hepatic systems as well as of human foetal intestine. Also, BMP-6 induces osteoblastic differentiation of pluripotent mesenchymal cells and is an autocrine stimulator of chondrocyte differentiation. The present study was carried out to investigate the effect of BMP-6 on human cobblestone-area-forming cells (CAFC), that represent the functional primitive repopulating haemopoietic stem cell in long-term bone marrow culture. Also, the effect of BMP-6 on marrow stroma production of interleukin-6, -11 and their common receptor gp130 that is expressed in haemopoietic stem cells and is indispensable for their proliferation and tri-lineage differentiation was examined. Moreover, the effect of BMP-6 on marrow stroma release of soluble adhesion molecule VCAM-1 mediating the primitive haemopoietic stem cell adhesion to marrow stroma was examined. The number of CAFC was significantly reduced after BMP-6 treatment from 88+/-10 per 10(5)cells in control cultures in a dose dependent manner to only 48+/-3 per 10(5)cells in 50 ng/ml BMP-6-treated cultures, P< 0.01. Quantitative ELISA measurement revealed 50 ng/ml BMP-6 was able to significantly reduce IL-6 and IL-11 production from marrow stroma, P< 0.01. Also, BMP-6 significantly increased soluble gp130 release by 7.4-fold in 50 ng/ml BMP-6-treated marrow stroma cultures. The profound rapid increase in this natural antagonist of human IL-6 cytokine family may reduce the gp130 signaling. Also, the soluble VCAM-1 released increased by two-fold in 50 ng/ml BMP-6-treated marrow stroma cultures. The marked increase in the soluble form may exert an antagonist effect on the function of VCAM-1 (ligand for VLA4). Recently, blocking the VLA4/VCAM-1 adhesion pathway was shown to mobilise haemopoietic CD34 positive cells in normal individuals. Also, we previously observed a significantly lower expression of VLA4 (CD49d) on G-CSF-mobilised blood CD34 positive cells than on bone marrow CD34 positive cells before mobilisation in the same normal donors. Since BMP are currently being used in clinical trials for bone repair and fracture healing, the present results suggest a possible role for BMP-6 in mobilising CD34 positive cells for transplantation. Further in vitro tests are required to evaluate this potential mobilising role of BMP-6 in human long-term bone marrow culture. PMID- 11401331 TI - Characterization of myogenic cell membrane: spontaneous formation of heterokaryotic myotubes between two different kinds of myoblasts. AB - In a previous study, it has been shown that presumptive mouse C2 myoblast cells are strongly resistant to HVJ (hemaglutinating virus of Japan, Sendai virus) mediated cell fusion, but do become capable of fusion upon differentiation. Quail myoblasts transformed with a temperature-sensitive mutant of Rous sarcoma virus (QM-RSV cells) also become more sensitive to HVJ-mediated cell fusion during differentiation. Investigations were undertaken to see whether heterokaryotic myotubes were formed spontaneously by co-culture of two different kinds of myogenic cells, QM-RSV cells and C2 cells. When both cells were committed to myotube formation, they spontaneously fused without HVJ on co-culture. On the other hand, when both or one of the cells were in the presumptive state, heterokaryons were not formed by co-culturing. Furthermore, committed QM-RSV cells did not fuse with non-myogenic cells. These results indicate that the membranes of myogenic cells change to become capable of fusion for myotube formation during differentiation. PMID- 11401333 TI - Utilization of amino acids in growing kidney proximal tubule cell cultures. AB - The growth of rat kidney proximal tubule cells was monitored continuously by the cellular incorporation of [methyl-(14)C] thymidine using scintillating microplates. The radioisotope had no effect on cell proliferation over a 5 day period, neither was it extensively converted to thymine. Leibovitz L-15 medium supplemented with bicarbonate proved a good growth medium and its high levels of carbohydrates and amino acids facilitated the appearance of intermediates in the cells' metabolism of additional radioactive amino acids. Kidney proximal tubule cells had a greater potential to process amino acids than BHK-21 cells. The utilization of amino acids by proximal tubule cells differed from that of other organs. The amino acids could be classified into three classes. Members of the first type were only used for protein synthesis (arginine, lysine, histidine and tyrosine). The second class of amino acids yielded only one or two metabolites (leucine and isoleucine), while the last type gave more than two metabolites (alanine, aspartate, glycine, methionine, proline and valine). PMID- 11401332 TI - Increased survival of mesothelial cells from the peritoneum in peritoneal dialysis fluid. AB - Peritonitis remains the most important factor in patient morbidity and technical failure associated with continuous ambulatory peritoneal dialysis (CAPD). In vitro examination of bacterial infection of cultured human peritoneal mesothelial cells (HPMC) is an attractive approach to the study of peritonitis in CAPD, yet there are few reports on this subject. Previous studies have shown two limitations: (i) cell cultures of HPMC lasted for days only when incubated in culture medium and (ii) short-term studies of <30 min were done in HPMC when incubated with peritoneal dialysis fluid (PDF). Human peritoneal mesothelial cells, maintained in a conventional single chamber culture system with PDF alone, were unable to survive more than 40 min. The present study was designed to prolong the viability of HPMC cultured in PDF, with the object of using cells under different conditions, such as that of simulating CAPD. HPMC were cultured using plastic microtiter plates, where they were grown to confluence and growth was arrested. PDF containing different concentrations of NaHCO3and human serum albumin was added. Cell viability after exposure for up to 24 h was measured by trypan blue, Cell Death Detection ELISA and Annex-V flow cytometry. The data confirmed the 'toxic' effect of PDF, with cell viability being <40% after 2 h incubation in 4.25% glucose in PDF. However, the survival time of HPMC increased significantly in 4.25% glucose PDF at a physiological pH and even further after the addition of human albumin. These experimental conditions simulating CAPD may allow future in vitro studies of mesothelial physiology and peritonitis related to CAPD treatment. PMID- 11401334 TI - Protein tyrosine kinase-dependent regulation of adenylate cyclase and phosphatidylinositol 3-kinase activates the expression of glial fibrillary acidic protein upon induction of differentiation in rat c6 glioma. AB - Glial fibrillary acidic protein (GFAP) is expressed upon cAMP-mediated induction of differentiation of glial progenitor cells into type II astrocytes. The protein is regulated by hormones, growth factors and cytokines but the signal transduction pathways involved in the regulation of GFAP expression are largely unknown. Specific protein kinase inhibitors were used to study their effect on the expression of GFAP in rat C6 glioma cells. Herbimycin A, a selective protein tyrosine kinase inhibitor, reduced GFAP mRNA and protein expression upon cAMP analog or beta-adrenergic receptor-mediated induction of differentiation. The latter inhibitor attenuated the elevation of cAMP by adenylate cyclase and abolished the activity of phosphatidylinositol 3-kinase (PI 3-K). These data indicate that GFAP expression is regulated by protein tyrosine phosphorylations, modulating the cAMP concentration and PI 3-K activity in C6 glioma cells. PMID- 11401335 TI - Low doses of biologically active substances: effects, possible mechanisms, and features. AB - Some substances can produce a response in biological systems at extraordinarily low concentrations. Often called the 'low' or 'small' dose effect, the meaning or definition of what is meant by 'low' in this context is discussed. For the purposes of this article the expression is taken to be a concentration substantially below the equilibrium dissociation constant of the effector-target complex, but extremely low concentrations (<10(-19) m) are excluded from consideration. The main features of very low or 'small' dose action are described. Possible mechanisms of the effects of very low doses are suggested. The main thrust of the paper concerns the explanation of bimodal and polymodal dose-effect curves. A general formal kinetic model for such curves and its application will be discussed. PMID- 11401336 TI - Possible role of fas antigen (CD 95) in human amniotic epithelial cell death: an in vitro study. AB - Treatment of human amniotic epithelial (HAE) cells with anti-Fas monoclonal antibody (CH 11) at 100 ng/ml or 1 microg/ml for 12 or 24 h increased necrotic cell death. Apoptotic cell death induced by this antibody was significantly increased, although far fewer cells underwent apoptosis, as determined by the TUNEL method. This study suggests that Fas antigen is an important mediator in HAE cell death. PMID- 11401337 TI - Early membrane depressions in hepatocytes cultured on Primaria supports. AB - The topography of spreading hepatocytes on positively charged Primaria plates was examined using scanning electron microscopy. The cells acquired a uniform morphology within 2 h. The spreading was rapid and the surface of the cells showed early prominent depressions or dips. The hepatocytes had either one or two of these structures which corresponded to the frequency of mononuclear and binuclear cells, respectively. The nuclear origin of the dips was strengthened after 6 h. They contained solid structures, whose number, size and shape were the same as nucleoli. The membrane dipping was independent of cell density and took place under conditions where phenotypic changes can occur. Kidney proximal tubule cells had no dips. Co-cultures of hepatocytes and kidney proximal tubule cells showed that the cell types behave differently. PMID- 11401338 TI - Tapping of Granular Packs: A Model Based on Local Two-Level Systems. AB - Two-level systems are known to be important for the low-temperature properties of glasses. We suggest here that they might explain some remarkable properties of powders under repeated tapping, as discovered by the Chicago group. Following the ideas of S. F. Edwards, the relevant variables here are (1) the volumes V(alpha) (V(beta)) occupied in the states alpha, beta (including distant reorganizations); (2) the magnitude B of the "activated volume" during a transition from alpha to beta; (3) the analog of temperatures, i.e., the compactivity (or free volume) v. Tapping induces alpha-->beta transitions, and these in turn reduce the compactivity. At low tapping strengths GammaGamma>Gamma(*)), the system freezes before reaching the alpha-beta equilibrium, and the density grows with the observed logarithmic law. At higher tapping strengths (Gamma0.1 microM), MT-III also antagonizes the trophic effect of Abeta(1-40) on cerebral cortical neurons in early cultures. Because only the fibrillar, SDS-resistant form of Abeta aggregates are thought to be neurotoxic, we analyzed and compared Abeta(1-40) aggregates formed in the presence and absence of MT-III using SDS PAGE. Results show that aggregates formed in the absence of MT-III are predominantly SDS-resistant whereas those formed in its presence are mostly SDS soluble. Neither MT-I nor -II exhibits any of the effects of MT-III. On the basis of these results, we propose that MT-III alleviates Abetas' neurotoxic effect by abolishing the formation of toxic aggregates of Abetas and that it may play a specific and important role in protecting the brain from the deleterious effects of Abetas. PMID- 11401475 TI - WISP-2: a serum-inducible gene differentially expressed in human normal breast epithelial cells and in MCF-7 breast tumor cells. AB - WISP-2 is a Wnt-1-induced signaling protein identified as a member of CCN growth factor family. A role for this molecule during tumorigenesis is suspected but remains unproven. Here we show that WISP-2 expression was undetectable, or minimally detectable, in nontransformed human mammary epithelial cells, but was overexpressed in MCF-7 cells. Expression of WISP-2 in MCF-7 cells was modulated by serum and correlated with the serum-induced MCF-7 tumor cell proliferation, suggesting that WISP-2 is serum responsive and may be a positive regulator of tumor cell proliferation. PMID- 11401476 TI - Differentiation in chronic myelogenous leukemia cell K562 by spongean sesterterpene. AB - Scalarane-type sesterterpenes, PHC-1-PHC-7, which have been isolated from a marine sponge, increased hemoglobin production in human chronic myelogenous leukemia cell line K562 at the concentration of 0.1-5 microg/ml. PHC-1, the major constituent, induced the expression of glycophorin A and the enucleation for K562 cells. These sesterterpenes were found to induce erythroid differentiation in K562 cells. In addition, PHC-1 induced G1 arrest of the cell cycle of K562 cells. PMID- 11401477 TI - Factor Xa-evoked relaxation in rat aorta: involvement of PAR-2. AB - Protease-activated receptor-2 (PAR-2) and/or effector cell protease receptor-1 (EPR-1) may mediate the direct cellular actions of coagulation factor Xa in some cultured cell lines. The present study examined if factor Xa could actually evoke relaxation through either of these receptor systems in isolated rat aorta. Factor Xa at 8.5-85 nM, like the PAR-2-activators trypsin and SLIGRL-NH(2), produced nitric oxide-dependent relaxation in the precontracted aortic rings. PAR-2 desensitization abolished relaxation responses to factor Xa as well as trypsin in the rings. The factor Xa interepidermal growth factor synthetic peptide L(83)FTRKL(88)(G)-NH(2), known to block factor Xa binding to EPR-1, failed to inhibit factor Xa-evoked relaxation in the preparations. Our findings provide evidence that factor Xa evokes relaxation by activating PAR-2, but independently of EPR-1, in the rat aorta. The factor Xa-PAR-2 pathway might thus contribute to the severe hypotension during sepsis, in which multiple coagulation factors including factor X would become activated and PAR-2 would be induced. PMID- 11401478 TI - In situ proteolytic digestion of inclusion body polypeptides occurs as a cascade process. AB - Misfolded proteins undergo a preferent degradation ruled by the housekeeping bacterial proteolytic system, but upon precipitation as inclusion bodies their stability dramatically increases. The susceptibility of aggregated polypeptides to proteolytic attack remains essentially unexplored in bacteria and also in eukaryotic cells. We have studied here the in vitro proteolysis of beta galactosidase fusion proteins by trypsin treatment of purified inclusion bodies. A cascade digestion process similar to that occurring in vivo has been observed in the insoluble fraction of the digestion reaction. This suggests that major protease target sites are not either lost or newly generated by protein precipitation and that the digestion occurs in situ probably on solvent-exposed surfaces of inclusion bodies. In addition, the sequence of the proteolytic attack is influenced by protein determinants other than amino acid sequence, the early digestion steps having a dramatic influence on the further cleavage susceptibility of the intermediate degradation fragments. These observations indicate unexpected conformational changes of inclusion body proteins during their site-limited digestion, that could promote protein release from aggregates, thus partially accounting for the plasticity of in vivo protein precipitation and solubilization in bacteria. PMID- 11401479 TI - Multivalent DNA-based immunization against hepatitis B virus with plasmids encoding surface and core antigens. AB - The immune response against hepatitis B surface and core antigens was evaluated by either coinoculation or independent intramuscular administration of pAEC compact DNA immunization vectors carrying their genes. The pAEC vectors bear just the essential elements for mammalian expression and bacterial amplification. Balb/c mice were immunized with 100 microg of each construct, either alone or in combination. In spite of lacking known immunostimulatory sequences (e.g., AACGTT), significant cellular (proliferative) and humoral immune responses were raised against both antigens. Coadministration of both plasmids maintained the immune response against the two antigens, without interference between them. Modulation of the antigen expression and further immune response, by using the Kozak's translation initiation sequence, was also analyzed. No differences due to its presence or absence were observed. PMID- 11401480 TI - MEK/ERK-mediated proliferation is negatively regulated by P38 map kinase in the human pancreatic cancer cell line, PANC-1. AB - Pancreatic carcinoma is characterized by a poor prognosis and lack of response to conventional therapy. The regulatory mechanisms for the rapid proliferation of pancreatic cancer cells and the particular aggressiveness of this cancer are still not fully understood. In mammalian cells, three MAPK families including ERK, JNK, and P38 MAPK have been characterized. ERK is known to play an important role in regulating pancreatic cancer cell proliferation. However, the role of P38 kinase in pancreatic cancer cell proliferation and its relationship with ERK are unclear. Using the specific P38 inhibitor, SB203580 we found that blockade of P38 MAP kinase significantly enhanced proliferation of the pancreatic cancer cell line, PANC-1 cell, in a concentration-dependent manner. In parallel with the stimulation of proliferation, blockade of P38 MAP kinase markedly induced MEK and ERK1/2 phosphorylation, indicating an interaction between MEK/ERK and P38 MAP kinase signaling. Clearly, the interaction between these kinase pathways does not involve transcription and translation because MEK/ERK was activated immediately upon SB203580 treatment. Furthermore, inhibition of the MEK/ERK cascade using the MEK inhibitor, PD098059 abolished SB203580-induced PANC-1 cell proliferation. From these results, we conclude that a MEK/ERK and P38 MAP kinase interaction is important for pancreatic cancer cell proliferation. Breaking the balance between these two signaling pathways will modify pancreatic cancer cell proliferation. PMID- 11401481 TI - Retention of glucose on oligosaccharide chains linked to the LH/hCG receptor prevents cell surface expression. AB - In the first step of asparagine-linked oligosaccharide chain maturation, terminal glucose residues are removed from the high mannose oligosaccharide core by glucosidases I and II. The role that glucose residues play in trafficking the luteinizing hormone/human chorionic gonadotropin (LH/hCG) receptor from the endoplasmic reticulum to the cell surface was investigated. Glucosidases I and II were inhibited by incubating 293 T cells transiently transfected with LH/hCG receptor cDNA with 5 mM 1-deoxynojirimycin (DNJ). DNJ treatment resulted in a marked reduction in cell surface [(125)I]hCG binding. Similar results were obtained from glucosidase I-deficient Lec 23 Chinese hamster ovarian (CHO) cells and wild-type CHO cells that were transiently transfected with LH/hCG receptor cDNA. Immunoprecipitation followed by Western blotting of transfected 293 T cells incubated in the presence or absence of 5 mM DNJ revealed that there is substantially less receptor in DNJ-treated cells than in control cells. These results show that the removal of glucose residues is necessary for trafficking the LH/hCG receptor to the cell surface. PMID- 11401482 TI - Perivascular cells regulate endothelial membrane type-1 matrix metalloproteinase activity. AB - Angiogenic stimuli selectively induced expression of membrane type-1 matrix metalloproteinase (MT1-MMP) transcripts and protein in human umbilical vein endothelial cells (HUVECs). Pro-MMP-2 activation was blocked by treatment with tissue inhibitor of metalloproteinases-2 (TIMP-2), but not by TIMP-1 or inhibitors of other proteinase classes. Anti-MT1-MMP antibodies abrogated recombinant pro-MMP-2 activation by plasma membranes, indicating that MT1-MMP is the main mediator of pro-MMP-2 activation in HUVECs. Cocultures of HUVECs with smooth muscle cells (SMC) or pericytes (PC) resulted in the suppression of HUVEC pro-MMP-2 activation. Treatment of A10 SMC conditioned media with a neutralising anti-TIMP-2 antibody prevented the suppression of HUVEC pro-MMP-2 activation. Inhibition of HUVEC MT1-MMP function by PC and SM3 SMC correlated with elevated TIMP-3 expression. Thus, perivascular supporting cells regulate the functions of proangiogenic MMPs elaborated by endothelial cells via selective expression of TIMPs. This interplay may be important for maintenance of blood vessel architecture and neovascularisation. PMID- 11401483 TI - Attenuation of tissue P-selectin and MCP-1 expression and intimal proliferation by AT(1) receptor blockade in hyperlipidemic rabbits. AB - Angiotensin-II (Ang-II) participates in the development and progression of atherosclerosis by activating type 1 (AT(1)) receptors. In vitro studies show that inflammatory factors, such as P-selectin and MCP-1, which can be upregulated by Ang-II, play an important role in atherogenesis. We examined the effect of AT(1) receptor blockade with losartan on the expression of P-selectin and MCP-1 in hypercholesterolemic rabbits. Since AT(1) receptor blockade is associated with feedback upregulation of renin-angiotensin system (RAS), we also examined alterations in plasma Ang-II levels by losartan therapy. Male NZW rabbits were fed regular chow (high cholesterol diet or high cholesterol diet + losartan 25 mg/kg/day). As expected, there was a marked intimal proliferation in association with increase in serum cholesterol (P < 0.001). In addition, there was a modest increase in plasma Ang-II levels (P < 0.05), and a significant increase in the expression of AT(1) receptors, P-selectin and MCP-1 in aortas of high cholesterol diet rabbits. Concurrent administration of losartan with high cholesterol diet attenuated aortic intimal proliferation induced a fivefold increase in plasma Ang II levels and caused a marked decrease in expression of P-selectin and MCP-1 without change in serum lipid levels and aortic AT(1) receptor expression. These observations in hypercholesterolemic animal models show that AT(1) receptor blockade is associated with modulation of P-selectin and MCP-1 expression concurrent with reduction in intimal proliferation. The rise in plasma Ang-II does not appear to limit the potential beneficial effect of losartan. PMID- 11401484 TI - Phospholipase A(2)-mediated fusion of neutrophil-derived membranes is augmented by phosphatidic acid. AB - Phosphatidic acid (PA), the product of phospholipase D (PLD) metabolism, is not only an important second messenger in neutrophil signal transduction but PA generation increases membrane fusogenicity. Following neutrophil stimulation, PA formation can be detected in azurophil, specific, and plasma membrane vesicle subcellular fractions, suggesting a potential role for PA formation in granule plasma membrane fusion. Neutrophil stimulation also activates phospholipase A(2) (PLA(2)) and the release of arachidonic acid. In vitro fusion of plasma membrane vesicles and specific granules with complex liposomes were dependent on PLA(2) (<10 microM Ca(2+)) while the presence of PA in the liposomes augmented the effects of PLA(2). Azurophil granules were extremely resistant to fusion (no fusion at 12 mM Ca(2+) even in the presence of PLA(2)). However, in the presence of both PA and PLA(2) fusion could be detected at <5 microM Ca(2+), suggesting a direct role for phospholipid metabolism in neutrophil degranulation. PMID- 11401485 TI - Ectopic expression of negative ARNT2 factor disrupts fish development. AB - ARNT factors are a cluster of bHLH-PAS factors that heterodimerize with other specific bHLH-PAS factors to mediate a wide range of biological responses. Previously, we obtained a truncated form of ARNT2-like factor, ARNT2A, from zebrafish, which encompasses the basic-helix-loop-helix and PAS A/B domains, but lacks a transactivation domain at its carboxyl end. Herein, we report another truncated ARNT2-like factor, ARNT2X, in zebrafish, which differs from ARNT2A at its N-terminal region. In cultured ZLE cells, transiently expressed ARNT2X and ARNT2A inhibited 2,3,7,8-TCDD-activated cyp1a1 transcription with different efficiencies. In the developing embryo, arnt2X mRNA was consistently expressed in the retinal and neural tube regions until the hatching stages, but it exhibited a more specific pattern at larval stages, including expression in the brain, eyes, hypothalamus, pharyngeal skeleton, heart, liver, pronephros duct, pectoral fin, and epithelial cells of the swim bladder. In contrast, arnt2A transcription diminished after hatching. Microinjecting a recombinant arnt2X-expression vector into fertilized eggs before cleavage stages caused severe defects in brain, eyes, pectoral fin, heart, and gut development. This suggests that the ARNT-mediated signal transduction pathways play important roles in fish tissue development. PMID- 11401486 TI - Induction of telomere shortening and replicative senescence by cryopreservation. AB - Cryopreservation can alter cellular function under certain conditions. In this report, we demonstrate the induction of cellular senescence after cells have been cryopreserved using a standard protocol. A retinal pigment epithelial cell line frozen at a specific freezing rate and subsequently thawed showed severely impaired proliferation compared to cells that were not cryopreserved. The induction of senescence was suggested by senescent associated beta-galactosidase activity and diminished bromo-deoxyuridine incorporation. A remarkable increase of single-strand DNA breaks in terminal restriction fragment (TRF) were found in cryopreserved cells immediately after thawing. The rate of mean TRF length shortening was accelerated after cryopreservation. Given this evidence, we hypothesize that cryopreservation may cause telomere shortening and cellular senescence under certain freezing conditions. PMID- 11401487 TI - Androgens regulate the mammalian homologues of invertebrate sex determination genes tra-2 and fox-1. AB - Androgens, like other steroid hormones, exert profound effects on cell growth and survival by modulating the expression of target genes. In vertebrates, androgens play a critical role downstream of the testis determination pathway, influencing the expression of sexually dimorphic traits. Among cells of the nervous system, motor neurons respond to trophic effects of androgen stimulation, with a subpopulation of spinal motor neurons exhibiting sexually dimorphic survival. To study the mechanisms of androgen action in these cells, we performed a subtractive screen for genes upregulated by androgen in a motor neuron cell line. We show androgen-inducible expression of two RNA-binding proteins that are the mammalian homologues of invertebrate sex determination genes. Androgens upregulate the expression of tra-2alpha, an enhancer of RNA splicing homologous to Drosophila tra-2, and promote redistribution of the protein from a diffuse to a speckled pattern within the nucleus. Similarly, androgens upregulate the expression of a novel gene homologous to Caenorhabditis elegans fox-1. These data indicate that androgens exert their effects, in part, by modulating the expression and function of genes involved in RNA processing, and identify homologues of invertebrate sex determination genes as androgen-responsive genes in mammals. PMID- 11401488 TI - Acute inhibition of proinsulin biosynthesis at the translational level by palmitic acid. AB - The effects of fatty acids on pancreatic beta cell are still controversial. Here, in order to determine whether free fatty acids acutely affect beta cell functions, we studied the effect of palmitic acid (PA) on proinsulin biosynthesis and insulin secretion using rat islets in vitro. Exposure of islets to PA for 1 h reduced glucose-stimulated proinsulin biosynthesis in a dose-dependent manner; in contrast, no change in insulin secretion was observed after 1 h incubation with PA. Furthermore, PA treatment did not cause any change of preproinsulin mRNA level during 1-h incubation period. Thus, our data indicate that PA primarily suppresses glucose-induced proinsulin biosynthesis within 1 h at the translational level. PMID- 11401489 TI - Effect of membrane fluidity on tyrosine kinase activity of reconstituted epidermal growth factor receptor. AB - Epidermal growth factor receptor (EGFR) was functionally reconstituted into liposome membrane. Triton X-100 was removed by Bio-beads SM-2. More than 80% of the reconstituted EGFR possessed right-side-out orientation with the EGF binding side facing the medium. The tyrosine kinase assay of the EGFR was carried out in the presence of the antibiotic alamethicin. The reconstituted EGFR tyrosine kinase was well activated by EGF. The influence of lipid composition on tyrosine kinase activity was investigated. Introduction of cholesterol into the dioleoylphophatidylcholine (DOPC) liposome membrane resulted in the decrease of tyrosine kinase activity. The tyrosine kinase activity of EGFR in distearylphosphatidylcholine liposome was much lower than that of EGFR-DOPC proteoliposome. Results indicated the importance of membrane fluidity on the apparent tyrosine kinase activity of reconstituted EGFR. PMID- 11401490 TI - PACT, a double-stranded RNA binding protein acts as a positive regulator for type I interferon gene induced by Newcastle disease virus. AB - Virus infection triggers innate responses to host cells including production of type I interferon (IFN). Since IFN production is also induced by treatment with poly(I:C), viral double-stranded (ds) RNA has been postulated to play a direct role in the process. In the present study, we investigated the effect of dsRNA binding proteins on virus-induced activation of the IFN-beta gene. We found that PACT, originally identified as protein activator for dsRNA-dependent protein kinase (PKR) and implicated in the regulation of translation, augmented IFN-beta gene activation induced by Newcastle disease virus. Concomitantly with the augmented activity of IFN-beta enhancer, increased activity of NF-kappaB and IRF 3 and IRF-7 was observed. For the observed effect, the dsRNA-binding activity of PACT was essential. We identified residues of PACT that interact with a presumptive target molecule to exert its function. Furthermore, PACT colocalized with viral replication complex in the infected cells. Thus the observed effect of PACT is novel and PACT is involved in the regulation of viral replication and results in a marked increase of cellular IFN-beta gene expression. PMID- 11401491 TI - Apoptosis in dissociation between DNA synthesis and cellular functions of activated hepatic stellate cells--a study with carbon tetrachloride-induced rat liver injury. AB - It is widely believed that DNA synthesis and expressions of smooth muscle alpha actin and TGF-beta are all together increased in activated hepatic stellate cells both in vitro and in vivo. Our previous reports disclosed that these increases did not always coexist under experimental conditions. Liver necrosis was induced in rats by oral administration of carbon tetrachloride. Hepatic stellate cells were isolated from these rats 2 days later. When these cells were cultured on plastic dishes for 3 days, they showed marked DNA synthesis and smooth muscle alpha actin and TGF-beta mRNA expressions assessed by (3)H-thymidine incorporation and Northern blotting, respectively. In the cells further cultured for 7 days, the DNA synthesis was decreased, whereas both smooth muscle alpha actin and TGF-beta mRNA expressions were increased, compared to the cells cultured for 3 days. The cells cultured for 10 days showed apoptotic nuclei positive for nick-end labeling, and DNA extracted from the cells revealed laddering patterns on agarose gels by electrophoresis. Apoptotic nuclei were also immunohistochemically found in stellate cells in the liver of rats 4 days after the intoxication. We conclude that apoptosis developed in activated hepatic stellate cells both in vitro and in vivo, and this may contribute to the discrepancy between DNA synthesis and cellular functions of the cells. PMID- 11401492 TI - Target gene therapy for alpha-fetoprotein-producing hepatocellular carcinoma by E1B55k-attenuated adenovirus. AB - Gene therapy using replication-competent adenovirus that selectively propagates in tumor cells may be an effective treatment for cancer. We developed an adenovirus that would be replication specific for hepatocellular carcinoma (HCC). Based on our finding that the E1B55k-deficient adenovirus was able to replicate in human primary hepatocytes, we therefore designed an adenovirus carrying E1A and attenuated E1B gene driven by the alpha-fetoprotein promoter (Adv-AFP-E1AdB), thus restricting the replication specificity in AFP-producing HCC. Replication of Adv-AFP-E1AdB in primary hepatocytes was practically negligible 4 days after infection. Although Adv-AFP-E1AdB replicated slowly in AFP-producing HCC, it efficiently destroyed HCC cells independent of their p53 status. Experiments were conducted in vivo using systemic administration of Adv-AFP-E1AdB and we observed tumor size reduction in nude mice having liver cancer. The use of replication competent adenovirus deficient of the E1B gene coupled to an AFP-targeting strategy may be a safe and efficacious treatment for HCC. PMID- 11401493 TI - Hormonally-regulated expression of voltage-operated Ca(2+) channels in osteocytic (MLO-Y4) cells. AB - Voltage operated calcium channels (VOCCs) are important in stimulus-response coupling in osteoblasts. We have investigated the expression of VOCCs in the mouse osteocyte cell line, MLO-Y4. Using the whole-cell patch clamp technique we were unable to detect any VOCC currents (n = 436) even in the presence of the L type VOCC agonist Bay K 8644 (n = 350). Reverse transcription polymerase chain reaction (RT-PCR), using primers to detect alpha(1C), alpha(1D), and alpha(1G) VOCC subunits (all of which are expressed in primary osteoblasts), did not generate detectable products with mRNA from MLO-Y4 cells. However, after treatment with physiological levels of hormones, VOCC alpha(1) subunit mRNAs were detected in MLO-Y4 cells. PTH, 17beta-estradiol, and dexamethasone-treatment induced expression of L-type (alpha(1C), alpha(1D)) subunit transcripts. ATP treatment induced expression of T-type (alpha(1G)) transcripts. Using whole-cell patch clamp we detected VOCC currents in 5-10% of cells after treatment. Current characteristics (L- or T-type) were consistent with the transcript expressed. PMID- 11401494 TI - A PDE6A promoter fragment directs transcription predominantly in the photoreceptor. AB - Rod photoreceptor cGMP phosphodiesterase (PDE6) is a key enzyme in the phototransduction cascade. Lines of transgenic mice were established to determine the spatial expression pattern directed by an upstream fragment of the PDE6A gene. RT-PCR analysis showed that three of four lines analyzed transcribed the transgene predominantly in the retina and weakly in brain. The line showing no transgene transcription did not contain an intact transgene. Transcription of the transgene in the three lines was found in retina and weakly in brain, but not in heart, kidney, liver, or lung. Transcripts were most predominant in the photoreceptors of the retina. These results demonstrate that a short segment of the upstream region of the PDE6A gene comprises a functional promoter that is most active in photoreceptors. PMID- 11401495 TI - Secreted Abeta does not mediate neurotoxicity by antibody-stimulated amyloid precursor protein. AB - Antibodies against APP, a precursor of Abeta deposited in Alzheimer's disease brain, have been shown to cause neuronal death. Therefore, it is important to determine whether Abeta mediates antibody-induced neurotoxicity. When primary neurons were treated with anti-APP antibodies, Abeta40 and Abeta42 in the cultured media were undetectable by an assay capable of detecting 100 nM Abeta peptides. However, exogenously treated Abeta1-42 or Abeta1-43 required >3 microM to exert neurotoxicity, and 25 microM Abeta1-40 was not neurotoxic. Glutathione ethyl-ester inhibited neuronal death by anti-APP antibody, but not death by Abeta1-42, whereas serum attenuated toxicity by Abeta1-42, but not by anti-APP antibody. Using immortalized neuronal cells, we specified the domain responsible for toxicity to be cytoplasmic His(657)-Lys(676), but not the Abeta1-42 region, of APP. This indicates that neuronal cell death by anti-APP antibody is not mediated by secreted Abeta. PMID- 11401496 TI - 4-Hydroxy-2-nonenal is a powerful endogenous inhibitor of endothelial response. AB - There is increasing evidence that lipid peroxidation is involved in many of the pathophysiologies associated with cardiovascular diseases, such as atherosclerosis and the long-term complications of diabetes. Among the products which originate from the peroxidation of cellular membrane lipids, 4-hydroxy-2 nonenal (HNE) is believed to be largely responsible for the cytopathological effects observed during oxidative stress in vivo. Here we found that HNE dramatically inhibited the expression of adhesion molecules induced by inflammatory stimuli in human aortic endothelial cells. The inhibition was found to be accompanied by a significant reduction of NF-kappaB activation followed by nuclear localization. This and the observation that the HNE treatment of the cells resulted in a rapid reduction of intracellular glutathione levels suggest that redox regulation of NF-kappaB may be involved in the modulation of the endothelial response by reactive aldehydes. PMID- 11401497 TI - A GrpE mutant containing the NH(2)-terminal "tail" region is able to displace bound polypeptide substrate from DnaK. AB - A key feature to the dimeric structure for the GrpE heat shock protein is the pair of long helices at the NH(2)-terminal end followed by a presumable extended segment of about 30 amino acids from each monomer. We have constructed a GrpE deletion mutant protein that contains only the unique tail portion (GrpE1-89) and another that is missing this region (GrpE88-197). Circular dichroism analysis shows that the GrpE1-89 mutant still contains one-third percent alpha-helical secondary structure. Using an assay that measures bound peptide to DnaK we show that the GrpE1-89 is able to lower the amount of bound peptide, whereas GrpE88 197 has no effect. Additionally, when the same peptide binding assay is carried out with the COOH-terminal domain of DnaK, the full-length GrpE and the two GrpE deletion mutants show little to no effect on peptide release. Furthermore, the GrpE88-197 mutant is able to enhance the off-rate of nucleotide from DnaK and the 1-89 mutant has no effect on the nucleotide release. Similar results of nucleotide release are observed with the NH(2)-terminal ATPase domain mutant of DnaK. The results presented show directly that there is interaction between the GrpE protein's "tail" region and the substrate COOH-terminal peptide binding domain of DnaK, although the effect is only fully manifest with an intact full length DnaK molecule. PMID- 11401498 TI - Fungicidal effect of antimicrobial peptide, PMAP-23, isolated from porcine myeloid against Candida albicans. AB - The antifungal activity and mechanism of a 23-mer peptide, PMAP-23, derived from pig myeloid was investigated. PMAP-23 displayed strong antifungal activity against yeast and mold. To investigate the antifungal mechanism of PMAP-23, fluorescence activated flow cytometry and confocal laser scanning microscopy were performed. Candida albicans treated with PMAP-23 showed higher fluorescence intensity by propidium iodide(PI) staining, which was similar to that of Melittin than untreated cells. Confocal microscopy showed that the peptide was located in the plasma membrane. The action of peptides against fungal cell membranes was examined by treating prepared protoplasts of C. albicans with the peptide and lipid vesicle titration test. The result showed that the peptide prevented the regeneration of fungal cell walls and induced release of the fluorescent dye trapped in the artificial membrane vesicles, indicating that the peptide exerts its antifungal activity by acting on the plasma lipid membrane. PMID- 11401499 TI - Rottlerin induces a transformed phenotype in human keratinocytes. AB - PKCdelta plays a fundamental role in cell cycle control. Consistent with its proposed tumour suppressor function, ras transfection of the human keratinocyte cell line HaCaT results in a loss of PKCdelta expression mediated by TGFalpha (Exp. Cell Res., 219, 299, 1995). To get more insight into the role of PKCdelta in keratinocytes, we investigated the effects of Rottlerin, a specific inhibitor of protein kinase Cdelta, in HaCaT cells. After Rottlerin treatment, HaCaT cells lost their cobble-stone morphology and displayed a spindle-shaped, fibroblastic phenotype. Additionally, the establishment of cell-cell contacts was prevented. This was caused by an internalization of E-cadherin and beta-catenin as assessed by immunofluorescence. A similar phenotype was observed in the presence of a neutralizing anti-E-cadherin antibody. Rottlerin-treated HaCaT cells proliferated like transformed cells in a three-dimensional cell culture system. We therefore conclude that PKCdelta is involved in mediating cell-cell contacts via E-cadherin and hence regulates differentiation in HaCaT cells. PMID- 11401500 TI - Organization of human ACAT-2 gene and its cell-type-specific promoter activity. AB - Acyl-CoA:cholesterol acyltransferase (ACAT) plays important roles in cellular cholesterol homeostasis. Two ACAT genes exist in mammals. We report here the genomic organization of human ACAT-2 gene and analysis of its promoter activity in various cell lines. The human ACAT-2 gene spans over 18 kb and contains 15 exons. Three transcription start sites and one poly(A) site are identified by the 5'/3'-RACE. In addition, the human ACAT-2 gene is linked to the insulin-like growth factor binding protein 6 (IGFBP-6) gene in a head-to-tail manner with a small intergenic region of about 1.2 kb. The 5'-flanking region of human ACAT-2 gene contains many potential cis-acting elements for multiple transcriptional regulatory factors but lacks TATA and CCAAT boxes. Using promoter-luciferase reporter assays, we demonstrate the transcriptional activity of ACAT-2 gene promoter is high in Caco-2 cells, especially after these cells become postconfluent and behave as intestinal enterocytes. PMID- 11401501 TI - The genes for anabolic 2-oxoglutarate: ferredoxin oxidoreductase from Hydrogenobacter thermophilus TK-6. AB - 2-Oxoglutarate: ferredoxin oxidoreductase (OGOR) of a thermophilic, chemolithoautotrophic hydrogen-oxidizing bacterium, Hydrogenobacter thermophilus TK-6, is involved in carbon dioxide fixation via a reductive TCA cycle. The enzyme is a heterodimer comprising subunits of 70 and 35 kDa. The structural genes for the subunits (korAB) were cloned with primers designed from N-terminal sequences of the purified enzyme. The korAB genes were followed by two open reading frames of unknown function (orf3 and orf4). KorA carried a binding motif for coenzyme A, and KorB carried binding motifs for Fe-S cluster and thiamine pyrophosphate. Active recombinant enzyme from korAB was produced in E. coli under the control of the lac promoter. orf3 and orf4 were not necessary for the expression of active enzyme in E. coli. The recombinant enzymes had high substrate specificity toward 2-oxoglutarate as in the case of the native enzyme purified from strain TK-6. PMID- 11401502 TI - Actinohivin, a novel anti-HIV protein from an actinomycete that inhibits syncytium formation: isolation, characterization, and biological activities. AB - Blocking human immunodeficiency virus (HIV) entry into target cells is an important goal of HIV and acquired immune deficiency syndrome (AIDS) therapies. We have searched for anti-HIV substances from microorganisms using a syncytium formation assay system constructed with HeLa/CD4/Lac-Z and HeLa/T-env/Tat cells. We discovered a novel anti-HIV protein that inhibits syncytium formation, designated as actinohivin, from a cultured broth of a soil isolate, actinomycete strain K97-0003. ESI mass spectrometry of actinohivin isolated from the culture filtrate showed an ion with molecular mass of 12,520.3 Da. The amino acid sequence was determined by N-terminal Edman degradation of the intact protein and peptide fragments formed by endoproteinase digestions. Actinohivin consists of a 114-amino-acid chain that exhibits internal sequence triplication. Actinohivin inhibited both T-cell and macrophage tropic syncytium formation, with IC(50) values of 60 and 700 nM, respectively, and the cytopathic effect of HIV-1(IIIB) in MT-4 cells, with IC(50) value of 230 nM. PMID- 11401503 TI - Hypoxia regulates avian cardiac Arnt and HIF-1alpha mRNA expression. AB - The aryl hydrocarbon receptor nuclear translocator (Arnt) and hypoxia-inducible factor (HIF)-1alpha mediate cellular responses to hypoxia. We investigated the ability of hypoxia to regulate Arnt and HIF-1alpha mRNA in the heart in vivo. We cloned avian Arnt, developed an in vivo model of chronic cardiac hypoxia, and measured expression of cardiac Arnt and HIF-1alpha mRNA by quantitative RT-PCR. Chronic hypoxic exposure (24 h to 15% O(2)) of day 9 chick embryos resulted in a 30-fold increase in covalent binding of (3)H-misonidazole, a hypoxic tissue marker, to cardiac tissue, and a 2-fold induction of cardiac inducible nitric oxide synthase mRNA, compared to normoxic controls. In this same model, cardiac Arnt mRNA expression decreased by 35%, while HIF-1alpha mRNA expression increased 400%. These data suggest that regulation of Arnt and HIF-1alpha mRNA expression may contribute to the physiological responses of the heart during prolonged hypoxia. PMID- 11401504 TI - Apolipoprotein E in macrophages and hepatocytes is eegraded via the proteasomal pathway. AB - Macrophage-derived apolipoprotein E (apoE) influences the susceptibility of the arterial wall to atherosclerosis. Previous studies have shown that production of apoE in these cells is regulated at a posttranscriptional level and is increased by inhibitors of proteasomal degradation. To further investigate this mechanism, we stably transfected RAW 264.7 macrophages and HepG2 cells with a construct overexpressing ubiquitin, the peptide targeting proteins to the proteasome, fused to an influenza virus hemagglutinin epitope tag. Ubiquitination of apoE was investigated by immunoprecipitation and Western blot analysis. In both cell types, apoE was ubiquitinated, and inhibition of proteasome function by lactacystin led to accumulation of ubiquitinated apoE. These studies provide strong evidence for proteasomal degradation of apoE in the two main cell types responsible for its production and indicate a possible new level of regulation of this important protein. PMID- 11401505 TI - Insulin activates phospholipase C-gamma1 via a PI-3 kinase dependent mechanism in 3T3-L1 adipocytes. AB - Previously we have shown that the insulin receptor and phospholipase C-gamma1 physically interact in the 3T3-L1 adipocyte cell line. In this study, we investigated the ability of insulin and PDGF to stimulate PLC-gamma1 enzyme activity as measured by PI-(4,5)P(2) hydrolysis. Both insulin and PDGF caused a rapid (<1 min) increase in PLC activity associated with the respective receptor. PDGF treatment resulted in a higher and more sustained stimulation of PLC-gamma1 activity compared to insulin (0.95 pmol/min/mg vs 0.68 pmol/min/mg). Furthermore, insulin and PDGF promoted increases in total cellular DAG, one of the products of PI-(4,5)P(2) hydrolysis. Insulin-stimulated PLC activity appears to be downstream of PI-3Kinase as the DAG increase was partially blocked by Wortmannin and addition of PI-(3,4,5)P(3) activated PLC-gamma1 in vitro. Inhibition of PLC using U73122 or an inhibitory peptide caused a decrease in insulin-stimulated 2 deoxyglucose transport and GLUT4 translocation that was rescued by the addition of OAG, a cell-permeable synthetic DAG. PMID- 11401506 TI - Protein kinase C-dependent supply of secretory granules to the plasma membrane. AB - To elucidate the mechanism for supplying secretory granules to the cell membrane, chromaffin cells isolated from the bovine adrenal medulla were observed by the evanescent wave microscopy after staining their granules with acridine orange. The secretory granules showed only a very small fluctuation, indicating their docking to the plasma membrane. The rate and range of movement increased greatly by application of botulinum toxin A or C. The number of secretory granules docked to the plasma membrane significantly decreased by botulinum toxin C. Conversely, the number increased greatly by activation of protein kinase C with phorbol 12,13 dibutyrate (PDBu). In the presence of an anti-actin reagent cytochalasin D, no increasing effect of PDBu on the number of docked granules was observed. While in the presence of an anti-mitotic reagent, colchicine, a clear increasing effect of PDBu was observed. The final step for supplying granules to the plasma membrane in endocrine cells is concluded to be mediated by a phosphorylation-dependent and actin-based transport system. PMID- 11401507 TI - Molecular dissection of the importin beta1-recognized nuclear targeting signal of parathyroid hormone-related protein. AB - Produced by various types of solid tumors, parathyroid hormone-related protein (PTHrP) is the causative agent of humoral hypercalcemia of malignancy. The similarity of PTHrP's amino-terminus to that of parathyroid hormone enables it to share some of the latter's signalling properties, but its carboxy-terminus confers distinct functions including a role in the nucleus/nucleolus in reducing apoptosis and enhancing cell proliferation. PTHrP nuclear import occurs via a novel importin beta1-mediated pathway. The present study uses several different direct binding assays to map the interaction of PTHrP with importin beta using a series of alanine mutated PTHrP peptides and truncated human importin beta1 derivatives. Our results indicate that PTHrP amino acids 83-93 (KTPGKKKKGK) are absolutely essential for importin beta1 recognition with residues 71-82 (TNKVETYKEQPL) additionally required for high affinity binding; residues 380-643 of importin beta1 are required for the interaction. Binding of importin beta1 to PTHrP is reduced in the presence of the GTP-bound but not GDP-bound form of the guanine nucleotide binding protein Ran, consistent with the idea that RanGTP binding to importin beta is involved in the release of PTHrP into the nucleus following translocation across the nuclear envelope. This study represents the first detailed examination of a modular, non-arginine-rich importin beta1 recognized nuclear targeting signal. PMID- 11401508 TI - Role of intercellular adhesion molecule 1 in indomethacin-induced ileitis. AB - Adhesion molecules have been implicated in the pathogenesis of inflammatory bowel diseases. We investigated their expression and contribution to leukocyte recruitment in experimental intestinal inflammation. Ileitis was induced in Sprague-Dawley rats by two injections of indomethacin (7.5 mg/kg), given 24 h apart. Endothelial intercellular adhesion molecule-1 (ICAM-1) expression was quantified using the dual radiolabeled monoclonal antibody technique and Mac-1 (CD11b/CD18) expression on leukocytes by flow cytometry. Leukocyte infiltration was monitored by tissue myeloperoxidase (MPO) activity. The first indomethacin injection induced a time- and site-dependent increase of ICAM-1 expression in ileal mucosa and muscularis. The second injection resulted in a reduction of ICAM 1 expression below constitutive levels whereas Mac-1 was upregulated. MPO changes paralleled lesion development over 48 h. ICAM-1 and MPO values were correlated for the first 24 h. Immunoneutralization of either ICAM-1 or Mac-1 attenuated mucosal injury. We conclude that (i) indomethacin-induced ileitis is associated with a temporally disassociated upregulation of ICAM-1 and (ii) despite a reduction in ICAM-1 after 24 h, ICAM-1, in concert with Mac-1, contributes to mucosal injury and leukocyte infiltration elicited by indomethacin. PMID- 11401509 TI - A significant participation of leukemia inhibitory factor in regulating the reproductive function in rats: a novel action of the pleiotropic cytokine. AB - Leukemia inhibitory factor (LIF) is a pleiotropic cytokine exhibiting diverse biological activities in various tissues and cell types. Accumulating evidence suggests a crucial role for LIF in regulating the hypothalamo-pituitary-adrenal axis, but information is limited regarding whether the cytokine also exerts a significant influence on other endocrine systems. In this study, we examined a possible involvement of LIF in the generation of ovarian steroid-induced luteinizing hormone (LH) and prolactin (PRL) surges in the rat. Experiments were performed on both normally fed and 3-day-fasted rats, which were ovariectomized and primed with estradiol and progesterone. From 11:00 to 18:00 h, blood was collected every 30 min to measure LH and PRL. All the following substances were given intracerebroventricularly at 11:00 h. Compared to control serum, undiluted and 5-times diluted preparations of anti-rat LIF serum caused a partial but significant suppression of LH surge to a similar extent. The two different concentrations of antibody also delayed the onset of PRL surge to a comparable degree. Fasted rats were devoid of significant surges of the hormones, while 1.0 and 3.0 nmol, but not 0.3 nmol, of recombinant murine LIF given to these animals led to a partial but significant recovery of both LH and PRL surges. This stimulatory potency of LIF on both hormones was already maximal at its 1.0-nmol dose. These results demonstrate for the first time a significant participation of LIF in the generation of LH and PRL surges in the rat. A novel action of the pleiotropic cytokine is reported herein. PMID- 11401510 TI - Serial analysis of gene expression in chronic hepatitis C and hepatocellular carcinoma. AB - Hepatitis C virus (HCV) causes chronic hepatitis C (CH-C) and is epidemiologically linked with the occurrence of hepatocellular carcinoma (HCC). To elucidate the comprehensive gene expression profiles of CH-C and HCC, serial analysis of gene expression (SAGE) libraries were made from CH-C and HCC tissues of a patient, and compared with a reported SAGE library of a normal liver (NL). Scatter plots of the distribution of tags from the HCC library exhibited the existence of many differentially expressed genes compared with those from the CH C and NL libraries. Up-regulation of IFN-gamma inducible genes and oxidative stress-inducible genes were identified in both the CH-C and HCC libraries, and some unpublished new genes were specifically up- or down-regulated in the HCC library. This genome-wide scanning study discloses the molecular portraits of CH C and HCC, and provides novel candidate genes that should help clarify the mechanism of hepatocarcinogenesis in the chronically HCV-infected liver. PMID- 11401511 TI - The Gal/GalNAc-specific lectin from the plant pathogenic basidiomycete Rhizoctonia solani is a member of the ricin-B family. AB - The lectin isolated from the phytopathogenic basidiomycete Rhizoctonia solani (RSA) is a homodimer of two noncovalently associated monomers of 15.5 kDa. RSA is a basic protein (pI > 9) which consists mainly of beta-sheets. A presumed relationship with ricin-B is supported by the sequence similarity between the N terminus of RSA and the N-terminal subdomain of ricin-B. Hydrophobic cluster analysis confirms that the N-terminus of both proteins has a comparable folding. RSA exhibits specificity towards Gal/GalNAc whereby the hydroxyls at the C3', C4', and C6' positions of the pyranose ring play a key role in the interaction with simple sugars. The carbohydrate-binding site of RSA apparently accommodates only a single sugar unit. Our results demonstrate an obvious evolutionary relationship between some fungal and plant lectins, but also provide evidence for the occurrence of a lectin consisting of subunits corresponding to a single subdomain of ricin-B. PMID- 11401512 TI - Inefficient processing of an olfactomedin-deficient myocilin mutant: potential physiological relevance to glaucoma. AB - Mutations in TIGR/MYOC (myocilin), a secretory protein of unknown function, have been recently linked to glaucoma. Most known mutations map to the C-terminus, an olfactomedin-like domain. We have previously shown that, in contrast to the wild type, a truncated form of myocilin lacking the olfactomedin domain is not secreted. In this study, we present evidence that the mutant protein is not correctly processed in the endoplasmic reticulum (ER) and accumulates into insoluble aggregates. In addition, we show that the presence of increasing amounts of mutant protein induces a fraction of the soluble, native myocilin to move to the insoluble fraction. Given the importance of such protein aggregates in the etiology of several aging-related diseases, we propose that olfactomedin defective mutants might contribute to the pathology of glaucoma through a mechanism involving intracellular accumulation of misfolded proteins. PMID- 11401513 TI - Aspirin inhibits matrix metalloproteinase-2 activity, increases E-cadherin production, and inhibits in vitro invasion of tumor cells. AB - Aspirin (acetylsalicylic acid) is a widely used anti-inflammatory drug. Recently, aspirin was shown to reduce the risk of development of cancer and mortality from it. Tumor metastasis is the most important cause of cancer death. The aim of the present study was to investigate if aspirin affects the invasion of cancer cells. Matrix metalloproteinases (MMPs) and cell adhesion molecules play important roles in the modulation of tumor invasion. Gelatin-based zymography assay showed that aspirin inhibited MMP-2 activity of SK-Hep-1 cancer cells. Matrigel-based chemoinvasion assay showed that aspirin inhibited in vitro invasion of SK-Hep-1 cancer cells. Aspirin treatment also increased the production of the cell adhesion molecule, E-cadherin, in Hep G2 cancer cells. Aspirin is a cyclooxygenase (COX) inhibitor. Treatment of cells with another COX inhibitor, sulindac, also inhibited MMP-2 activity and increased E-cadherin production of cells. These results indicate that aspirin can modulate both MMP-2 and E-cadherin production and therein may possess antimetastatic effect. PMID- 11401514 TI - Chemoenzymatic synthesis of neoglycoproteins using transglycosylation with endo beta-N-acetylglucosaminidase A. AB - A novel chemoenzymatic approach to synthesize neoglycoproteins containing high mannose-type oligosaccharides is described. p-Isothiocyanatophenyl-beta-d glucopyranoside (Glc-ITC) was transferred to the reducing end of the high-mannose type oligosaccharides using a transglycosylation activity of endo-beta-N acetylglucosaminidase A (Endo-A). A novel oligosaccharide, Man(6)GlcNAc-Glc-ITC, was synthesized as a coupling reagent for lysyl and N-terminal residues of the protein moiety. The neoglycoconjugate was coupled with several nonglycosylated proteins such as ribonuclease A, lysozyme, and alpha-lactalbumin. Between one and four high-mannose-type oligosaccharides were incorporated per molecule of these proteins. This method should be very useful for the synthesis of neoglycoproteins with homogeneous high-mannose-type oligosaccharides. PMID- 11401515 TI - Detection of aquaporin-2 in the plasma membranes of oocytes: a novel isolation method with improved yield and purity. AB - Aquaporin-2 (AQP2) water channel mutations cause autosomal recessive and dominant nephrogenic diabetes insipidus (NDI). Expressed in oocytes, a mutant in dominant (AQP2-E258K), but not in recessive (AQP2-R187C), NDI conferred a specific dominant-negative effect on wild-type (wt) AQP2 water permeability (Pf) only at low expression levels. Since at these levels, the yield of conventional-isolated plasma membranes was too low, an improved technique to semiquantify AQP2 in the plasma membrane was needed. Antibodies against the C-loop of AQP2 were not applicable since they were unspecific and introduction of a tag into this loop caused misfolding and ER retardation. Membrane-impermeable biotin analogues turned out to label intracellular AQP2 proteins. Therefore, a method has been developed which generates a high yield of nearly pure plasma membranes, which enables semiquantification of plasma membrane proteins expressed at low levels in oocytes. Our new method allows for phenotype-genotype correlation studies in a wide range of channelopathies. PMID- 11401516 TI - Anionic phospholipids are involved in membrane targeting of PI 3-kinase gamma. AB - Phosphoinositide 3-kinases (PI 3-kinases) have critical roles in diverse cellular signaling processes and in protein trafficking. In contrast to the class I PI 3 kinases alpha, beta, and delta which bind via src homology 2 (SH2) domains of adaptor proteins to tyrosine kinase receptors, the mechanism of recruitment of the PI 3-kinase gamma to membranes is unknown. We report in vitro experiments using immobilized proteins and small unilamellar vesicles which suggest an involvement of anionic phospholipids in membrane association of PI 3-kinase gamma. Furthermore we provide evidence that the enzyme displays beside the catalytic center a phospholipid binding domain which is essential for enzyme activity. PMID- 11401517 TI - Both platelet-derived growth factor receptor (PDGFR)-alpha and PDGFR-beta promote murine fibroblast cell migration. AB - Cell motility plays a critical role for many physiological and pathological processes including wound healing, fibrosis, angiogenesis, and tumor metastasis. Platelet-derived growth factor (PDGF) is among the most potent stimuli for mesenchymal cell migration. The PDGF B-chain homodimer PDGF BB activates both alpha- and beta-receptor subunits (alpha-PDGFR and beta-PDGFR), and promotes cell migration in many cell types including fibroblasts and smooth muscle cells. PDGF A chain homodimer PDGF AA activates alpha-PDGFR only, and its role for cell migration is still debatable. PDGF BB, but not PDGF AA, induces smooth muscle cell migration. Interestingly, alpha-PDGFR was shown to antagonize beta-PDGFR induced smooth muscle cell migration. In the present study, we investigated the role of alpha-PDGFR and beta-PDGFR in PDGF-mediated cell migration of murine fibroblasts (NIH 3T3). Unlike smooth muscle cells, both PDGF AA and PDGF BB promoted NIH 3T3 cell migration. The effect of PDGF BB activation of beta-PDGFR alone for cell migration was examined using previously established NIH 3T3 clones in which alpha-PDGFR signaling is inhibited by a dominant-negative alpha-PDGFR, or an antisense construct of alpha-PDGFR. PDGF BB activation of beta-PDGFR alone was sufficient to induce cell migration, but the efficiency was significantly lower compared to PDGF activation of both receptors. These results showed that both alpha- and beta-PDGFRs promote fibroblast cell migration and their effects are additive. Taken together, we propose that cell-type specific alpha-PDGFR signaling is critical for regulation of mesenchymal cell migration in response to PDGF isoform, whereas beta-PDGFR mainly promotes cell migration. PMID- 11401518 TI - Identification of putative downstream genes of Oct-4 by suppression-subtractive hybridization. AB - As a step toward understanding how toti/pluripotence is maintained by Oct-4, we have first constructed a cell model with differentially expressed Oct-4 in embryonic stem cells, and then used suppression-subtractive hybridization (SSH) method to identify the downstream genes of Oct-4. Among the 384 clones we screened, 40 clones were detected as differentially expressed genes with colony hybridization, and 13 clones were confirmed as the putative downstream genes of Oct-4 by Northern blot analysis. Sequencing showed 12 different genes, 8 known genes (Oct-4, Rex-1, Sox-2, Creatine kinase B, Makorin 1, Importin beta, Histone H2A.Z, Ribosomal protein S7) and 4 new genes. Except Oct-4 and Rex-1, the other genes have not been reported to be regulated by Oct-4. These results showed that SSH provides a very efficient means to identify the downstream genes of transcription factor. Some known genes identified may provide new insight of the function of Oct-4 in stem cells. PMID- 11401519 TI - Mice with only rat mtDNA are required as models of mitochondrial diseases. AB - We examined the possibility of generation of mice expressing mitochondrial dysfunction by introduction of exogenous mtDNA from different species using mouse mtDNA-less (rho(0)) cells as mtDNA recipients. For determination of how genetically distant species of mtDNA could replicate in cells with only the mouse nuclear genome, we introduced mtDNA of the Syrian hamster (Mesocricetus auratus) into mouse rho(0) cells, and found that its replication was not sufficient to propagate to following generations, probably due to significant incompatibility between mouse-nuclear and Syrian hamster-mitochondrial genomes. On the other hand, rat mtDNA, which propagated stably and expressed mitochondrial dysfunction in mouse cells, also disappeared rapidly by exogenous introduction of mouse mtDNA, suggesting that mouse mtDNA in mouse cells must be excluded completely before introduction of rat mtDNA for generation of mice with rat mtDNA as mitochondrial disease models. PMID- 11401520 TI - alphaA- and alphaB-crystallins protect glucose-6-phosphate dehydrogenase against UVB irradiation-induced inactivation. AB - alpha-Crystallin, a major eye lens protein, has been shown to function like a molecular chaperone by suppressing the aggregation of other proteins induced by various stress conditions. Ultraviolet (UV) radiation is known to cause structural and functional alterations in the lens macromolecules. Earlier we observed that exposure of rat lens to in vitro UV radiation led to inactivation of many lens enzymes including glucose-6-phosphate dehydrogenase (G6PD). In the present paper, we show that alpha-crystallin (alphaA and alphaB) protects G6PD from UVB irradiation induced inactivation. While, at 25 degrees C, there was a time-dependent decrease in G6PD activity upon irradiation at 300 nm, at 40 degrees C there was a complete loss of activity within 30 min even without irradiation. The loss of activity of G6PD was prevented significantly, if alphaA- or alphaB-crystallin was present during irradiation. At 25 degrees C, alphaB crystallin was slightly a better chaperone in protecting G6PD against UVB inactivation. Interestingly, at 40 degrees C, alphaA- and alphaB-crystallins not only prevent the loss of G6PD activity but also protect against UVB inactivation. However, alphaA- and alphaB-crystallins were equally efficient at 40 degrees C in protecting G6PD. PMID- 11401521 TI - Dual function of troglitazone in ICAM-1 gene expression in human vascular endothelium. AB - Our previous work has shown that troglitazone (an antidiabetic, thiazolidione drug and a synthetic ligand for peroxisome proliferator-activated receptor gamma, PPARgamma) stimulated basal level of intercellular adhesion molecule-1 (ICAM-1) protein expression in the absence of cytokine stimulation in human vascular endothelial cells. In this study, we examine the molecular mechanism of troglitazone on the basal and TNFalpha-induced ICAM-1 gene expression. Activation of transcription factors, NF-kappaB and AP-1 proteins, known to regulate ICAM-1 gene expression upon external stimulators, was examined. In human vascular endothelial cells (ECV304 cells), troglitazone inhibited TNFalpha-induced ICAM-1 gene expression by suppressing NF-kappaB/DNA binding activity, NF-kappaB transcriptional responses, c-Fos mRNA and protein levels via a ligand-dependent, PPARgamma-activated manner. In contrast, both troglitazone (at 10 microM) and 15 deoxy-Delta(12,14)-prostaglandin J(2) (15d-PGJ(2), at 15 microM), a natural ligand for PPARgamma, induce c-Jun phosphorylation by activation of c-Jun N terminal kinase (JNK) through a posttranslational regulation of c-Jun activity, therefore increasing AP-1/DNA binding activity and transcriptional responses as results of increasing basal ICAM-1 gene expression. These findings suggest dual function of troglitazone in the modulation of both basal and stimulated ICAM 1gene expression in human vascular endothelial cells. PMID- 11401522 TI - Expression of anaphase-promoting complex 5 in balloon angioplasty-injured rat carotid arteries and mitogen-stimulated human vascular smooth muscle cells. AB - We utilized differential display to identify differentially expressed mRNAs induced by balloon angioplasty injury. A recently described subunit of the anaphase-promoting complex, APC5, is not expressed in uninjured rat carotid arteries. Expression of APC5 mRNA is detectable as early as 1 day post-injury, reached maximal levels by 3-7 days, and declined by 14 days post-injury. APC5 is not expressed in serum-starved cultured human vascular smooth muscle cells (VSMC), but is strongly induced by mitogenic factors, and to a lesser extent by nonmitogenic cytokines. The kinetics of APC5 expression is associated with cell cycle progression, and corresponds with PCNA expression in PDGF-stimulated human VSMC. Interestingly, APC5 is expressed in most human tissues examined, regardless of their proliferative state. These data are the first description of the cytokine-inducible expression of APC5 and suggest that expression of this gene may represent an important event in the pathogenesis of vascular proliferative diseases. PMID- 11401523 TI - CaT1 expression correlates with tumor grade in prostate cancer. AB - Ca(2+) signaling is important for growth and survival of prostatic carcinoma (PCa) cells. Here we report that the gene for CaT1, a channel protein highly selective for Ca(2+), is expressed at high levels in human PCa and in the LNCaP PCa cell line. CaT1 mRNA levels were elevated in PCa specimens in comparison to benign prostatic hyperplasia (BPH) specimens and positively correlated with Gleason grade in a PCa series. CaT1 mRNA was suppressed by androgen and was induced by a specific androgen receptor antagonist in LNCaP cells, suggesting that the gene is negatively regulated by androgen. These findings are the first to implicate a Ca(2+) channel in PCa progression and suggest that CaT1 may be a novel target for therapy. PMID- 11401524 TI - Expression of IL-17 mRNA in ovarian cancer. AB - IL-17 is considered as a proinflammatory cytokine. We have demonstrated IL-17 is an angiogenic factor and promotes tumor growth in murine tumor models. In this report, we investigated the expression of IL-17 mRNA by RT-PCR and the relationship between IL-17 expression and microvascular density in ovarian cancer. IL-17 mRNA was expressed in 11 (64.7%) of 17 ovarian cancer. And the average number of blood vessels observed in IL-17 positive tumors (173.4 +/- 55.1/mm(2)) was significantly higher than that in negative tumors (107.7 +/- 57.8/mm(2)). These results indicated IL-17 is expressed in a considerable proportion of ovarian cancer and promotes tumor angiogenesis. There was no significant relationship between IL-17 expression and clinicopathologic parameters. PMID- 11401525 TI - Requirement of the basic region of N-WASP/WAVE2 for actin-based motility. AB - WASP family proteins activate nucleation by the Arp2/3 complex, inducing rapid actin polymerization in vitro. Although the C-terminal portion of WASP family proteins (VCA) activates nucleation by the Arp2/3 complex in pure systems, we find that this fragment lacks activity in cell extracts. Thus, polystyrene beads coated with VCA did not move in brain cytosol, while beads coated with N-WASP or WAVE2 did move. The basic clusters between the WH1 domain and the CRIB domain of N-WASP were critical for movement since beads coated with N-WASP or WAVE2 constructs missing the basic clusters (Delta basic) also did not move. Furthermore, VCA and N-WASP/WAVE2 Delta basic constructs were much less able than wild-type N-WASP and WAVE2 to induce actin polymerization in cytosol. All of the proteins, with or without the basic domain, were potent activators of nucleation by purified Arp2/3 complex. PMID- 11401526 TI - Activin A induces expression of rat Sel-1l mRNA, a negative regulator of notch signaling, in rat salivary gland-derived epithelial cells. AB - We previously established a rat submandibular gland (SMG)-derived epithelial cell line (RSMG-1) to study the mechanism of morphogenesis in salivary gland development and regeneration. We found that activin A regulated the branching morphogenesis of RSMG-1 cells, suggesting that it is involved in SMG morphogenesis. We used a subtraction cloning procedure with activin-A-treated and untreated RSMG-1 cells to identify activin-A-induced genes. One of the genes detected encoded a rat homologue of Sel-1l (rSel-1l). rSel-1l is a mammalian homologue of C. elegans sel-1, which is a negative regulator of Notch signaling. In this study, we confirmed that activin A induces rSel-1l mRNA expression in RSMG-1 cells, and that rSel-1l is expressed in SMG acinar cells. These results suggest that activin A regulates the differentiation of RSMG-1 cells to acinar cells. PMID- 11401527 TI - FZD4S, a splicing variant of frizzled-4, encodes a soluble-type positive regulator of the WNT signaling pathway. AB - Frizzled-1 (FZD1)-FZD10 are seven-transmembrane-type WNT receptors, and SFRP1 SFRP5 are soluble-type WNT antagonists. These molecules are encoded by mutually distinct genes. We have previously isolated and characterized the 7.7-kb FZD4 mRNA, encoding a seven-transmembrane receptor with the extracellular cysteine rich domain (CRD). Here, we have cloned and characterized FZD4S, a splicing variant of the FZD4 gene. FZD4S, corresponding to the 10.0-kb FZD4 mRNA, consisted of exon 1, intron 1, and exon 2 of the FZD4 gene. FZD4S encoded a soluble-type polypeptide with the N-terminal part of CRD, and was expressed in human fetal kidney. Injection of synthetic FZD4S mRNA into the ventral marginal zone of Xenopus embryos at the 4-cell stage did not induce axis duplication by itself, but augmented the axis duplication potential of coinjected Xwnt-8 mRNA. These results indicate that the FZD4 gene gives rise to soluble-type FZD4S as well as seven-transmembrane-type FZD4 due to alternative splicing, and strongly suggest that FZD4S plays a role as a positive regulator of the WNT signaling pathway. PMID- 11401528 TI - Molecular characterization of a novel glycoprotein hormone G-protein-coupled receptor. AB - We report the molecular characterization of a novel G-protein-coupled receptor, GPR48, that resembles proteins in the glycoprotein hormone receptor family. The full-length human GPR48 cDNA is comprised of 951 amino acids. The large extracellular amino terminus of 538 residues is composed of seventeen leucine rich repeats (LRR). The genomic structure of GPR48 has several features in common with genes in the glycoprotein hormone receptor family. Analogous to these receptors, most of the LRR are encoded on single small exons, and the last exon encodes the seven transmembrane segments. The complete gene spans more than 60 kb with 18 exons and 17 introns. Northern blot analysis demonstrated high expression of GPR48 in the adult human pancreas, with moderate levels of expression in placenta, kidney, brain, and heart. Additionally, this receptor is expressed as early as 7 days post coitus in the mouse, indicating its potential involvement in development. PMID- 11401529 TI - Lysophosphatidic acid- and Gbeta-dependent activation of Dictyostelium MAP kinase ERK2. AB - Exogenous lysophosphatidic acid (LPA) has been shown to evoke a chemotactic response in aggregative cells of the social amoeba Dictyostelium discoideum. In this paper, we demonstrate that extracellular LPA is also able to induce activation of mitogen-activated protein (MAP) kinase DdERK2 (extracellular signal regulated kinase 2) in these cells. This activation is independent of cyclic AMP receptors, yet fully dependent on the single Gbeta subunit, hinting to the presence of functional heptahelical LPA receptors in a primitive eukaryote. We did not observe LPA-dependent cyclic GMP accumulation, which suggests that the pathways for LPA-induced and "classical" chemotaxis of D. discoideum cells are substantially different. PMID- 11401530 TI - CaM-kinase II dephosphorylates Thr(286) by a reversal of the autophosphorylation reaction. AB - Autophosphorylation of CaM-kinase II produces a form of the enzyme not requiring Ca(2+)/calmodulin for sustained activity. We report that autophosphorylated CaM kinase II dephosphorylates itself in the presence of ADP (termed autodephosphorylation). The dephosphorylation was unaffected by phosphatase inhibitors and was nucleotide specific, occurring with ADP but not with AMP, GTP, or GDP. (32)P-ATP was produced when ADP was added to (32)P-labeled CaM-kinase II, indicating that the enzyme was undergoing dephosphorylation through a reversal of the autophosphorylation reaction. ATP addition also produced loss of (32)P from the autophosphorylated enzyme while maintaining the kinase in a phosphorylated state. This indicates that the enzyme was undergoing cycles of autophosphorylation and dephosphorylation in the activated state. Autothiophosphorylated CaM-kinase II was resistant to autodephosphorylation. Site directed mutants were used to show that Thr(286) was the predominant site dephosphorylated. Additionally, CaM-kinase II composed of beta subunits exhibited autodephosphorylation. Ca(2+)/CaM-independent activity expressed by the autophosphorylated alpha and beta holoenzymes was reversed following autodephosphorylation. PMID- 11401531 TI - Significance of ecto-cyclase activity of CD38 in insulin secretion of mouse pancreatic islet cells. AB - Cyclic ADP-ribose (cADPR), a product of CD38, has a second messenger role for in intracellular Ca(2+) mobilization from microsomes of pancreatic islets as well as from a variety of other cells. ADP-ribosylation of CD38 by ecto-mono ADP ribosyltransferase in activated T cells results in apoptosis as well as inactivation of its activities. We, therefore, examined the effect of ADP ribosylation of CD38 in mouse pancreatic islet cells. NAD-dependent inactivation and ADP-ribosylation of CD38, intracellular concentrations of cADPR and Ca(2+), and insulin secretion were measured following incubation of mouse pancreatic islet cells with NAD. ADP-ribosylation of CD38 inactivated its ecto-enzyme activities, and abolished glucose-induced increase of cADPR production, intracellular concentration of Ca(2+), and insulin secretion. Taken together, ecto-cyclase activity of CD38 to produce intracellular cADPR seems to be indispensable for insulin secretion. PMID- 11401532 TI - Purification and structural analysis of the hepatitis B virus preS1 expressed from Escherichia coli. AB - The preS1 of hepatitis B virus (HBV) is located at the outermost part of the envelope protein and possesses several functionally important regions such as hepatocyte receptor-binding site and virus-neutralizing epitopes. As the first step to understand the structure-function relationship for the preS1 antigen, we have purified the preS1 and performed its structural characterization by circular dichroism (CD) spectroscopy. The preS1 was purified to near homogeneity from bacterially expressed glutathione S-transferase (GST)-preS1 fusion protein by two step purification, affinity chromatography on glutathione-agarose column, and cation-exchange chromatography on Mono S column. The CD analysis showed that the purified preS1, which was largely unstructured in aqueous solution, acquired a significant (16%) alpha-helical structure when analyzed in 50% trifluoroethanol or 20 mM SDS. The results suggest that the preS1 assumes a mainly unstructured conformation and may form induced secondary structures upon binding to target proteins or under hydrophobic environment. PMID- 11401533 TI - Thiouracil antithyroid drugs as a new class of neuronal nitric oxide synthase inhibitors. AB - Two established antithyroid drugs, 6-propyl-2-thiouracil and 6-methyl-2 thiouracil, as well as S-methylthiouracil, are shown to be competitive inhibitors of nitric oxide synthase (NOS) (K(I) values ranging from 14 to 60 microM), with moderate selectivity for the neuronal isoform. Other thioureylene and thioamide containing heterocyclic systems proved virtually ineffective as NOS inhibitors. Besides offering novel useful leads for inhibitor design as well as to probe the active site of neuronal NOS, the results of this study may have interesting implications in relation to the antithyroid activity of thiouracils and their possible neurological effects. PMID- 11401534 TI - Phosphate decreases osteoclastogenesis in coculture of osteoblast and bone marrow. AB - Alkaline phosphatase (ALP) activity increases dramatically during osteoblast maturation, a phenomenon that is related to calcification of teeth and bone. Although the relation between ALP and calcification is widely known, there is a paucity of date relating ALP and osteoclast formation. Very recently, we showed that osteoblast maturation suppresses osteoclast formation. However, the relation between osteoblast maturation and osteoclastogenesis remains unclear. In the present study, we examined the effect of extracellular phosphate on osteoclastogenesis. As osteoblasts matured, osteoclast formation decreased, and ALP activity and inorganic phosphate in extracellular matrix increased. Inorganic phosphate in extracellular matrix and extracellular phosphate was suppressed osteoclast formation. These results suggest that phosphate released by ALP may be related not only to calcification but also to suppression of osteoclastogenesis. PMID- 11401535 TI - Alternative splicing of novel exons of rat heart-type fructose-6-phosphate 2 kinase/fructose-2,6-bisphosphatase gene. AB - Three C-terminal sequences (VSIPVV, VCKWT, and LTLLS) of rat heart-type fructose 6-phosphate 2-kinase/fructose-2,6-bisphosphatase have been reported. To elucidate the mechanism of generating the heterogeneity of the enzyme, we carried out reverse transcriptase PCR using three pairs of specific primers. The existence of mRNAs encoding VSIPVV and LTLLS was confirmed but not that of VCKWT. In addition to these cDNAs, we found four novel mRNAs that encode the C-termini of the enzyme. The genomic sequence reveals that the heterogeneity is generated by alternative splicing of exon 15 and four novel exons (exon 16a-d). PMID- 11401536 TI - A pH-sensitive interaction of troponin I with troponin C coupled with strongly binding cross-bridges in cardiac myofilament activation. AB - Slow skeletal muscle troponin I (ssTnI) expressed predominantly in perinatal heart confers a marked resistance to acidic pH on Ca(2+) regulation of cardiac muscle contraction. To explore the molecular mechanism underlying this phenomenon, we investigated the roles of TnI isoforms (ssTnI and cardiac TnI (cTnI)) in the thin filament activation by strongly binding cross-bridges, by exchanging troponin subunits in cardiac permeabilized muscle fibers. Fetal cardiac muscle showed a marked resistance to acidic pH in activation of the thin filament by strongly binding cross-bridges compared to adult muscle. Exchanging ssTnI into adult fibers altered the pH sensitivity from adult to fetal type, indicating that ssTnI also confers a marked resistance to acidic pH on the cross bridge-induced thin filament activation. However, the adult fibers containing ssTnI or cTnI but lacking TnC showed no pH sensitivity. These findings provide the first evidence for the coupling between strongly binding cross-bridges and a pH-sensitive interaction of TnI with TnC in cardiac muscle contraction, as a molecular basis of the mechanism conferring the differential pH sensitivity on Ca(2+) regulation. PMID- 11401537 TI - Role of phospholipase C-gamma1 in insulin-like growth factor I-induced muscle differentiation of H9c2 cardiac myoblasts. AB - Insulin-like growth factor-I (IGF-I) regulates muscle differentiation through phosphatidylinositol 3-kinase (PI 3-kinase). Also it was recently reported that PI 3-kinase is involved in the activation of phospholipase C-gamma1 (PLC-gamma1). We investigated whether PLC-gamma1 therefore plays a role in IGF-I-induced muscle differentiation using H9c2 rat cardiac myoblasts as a model. IGF-I was able to activate PLC-gamma1 via both PI 3-kinase-dependent and tyrosine phosphorylation dependent mechanisms in this model. However, PI 3-kinase appeared to play a more important role than tyrosine phosphorylation in IGF-I activation of PLC-gamma1. In addition, PLC-gamma1 activation was independent of Akt/protein kinase B (Akt/PKB). Importantly, PLC-gamma1 was involved in IGF-I-induced muscle differentiation in parallel with Akt/PKB. Taken together, these results suggest that IGF-I regulation of muscle differentiation is dependent on the activation of PLC-gamma1 and Akt/PKB, both of which are downstream mediators of PI 3-kinase. PMID- 11401538 TI - The 5' flanking region of the human bone morphogenetic protein-7 gene. AB - We describe here the cloning and characterization of the 5' flanking region of the human Bone Morphogenetic Protein-7 (BMP-7) gene from a 3.3 kb genomic DNA fragment. Functional analysis by transient transfection using the luciferase reporter gene indicated that this region had a low basal promoter activity in human Wilm's tumor derived renal (G401) and rat osteoblast (ROS 17/2.8) cell lines. Sequential deletion analysis of the promoter revealed sequences whose presence correlated with decreased expression of the reporter gene. Coexpression of transcription factors involved in epithelial/mesenchymal interactions during kidney and eye development dramatically stimulated the expression of the reporter gene from the putative BMP-7 promoter. Finally, a subset of agents that upregulated the expression of the reporter gene from the cloned promoter were also shown to increase the expression of the endogenous BMP-7 in G401 and ROS cell lines in vitro. PMID- 11401539 TI - Isolation and characterization of acetoacetyl-CoA thiolase gene essential for n decane assimilation in yeast Yarrowia lipolytica. AB - Yarrowia lipolytica is a yeast which can utilize n-alkane as a sole carbon source. We isolated a Y. lipolytica peroxisomal acetoacetyl-CoA thiolase gene, PAT1, by complementation of a mutant that cannot utilize n-decane as a sole carbon source. We found that the putative PAT1 product had conserved features of peroxisomal acetoacetyl-CoA thiolase. We showed that the PAT1 disruptant was not able to grow on n-decane, and that n-decane-inducible acetoacetyl-CoA thiolase activity largely depended on PAT1. The original mutant carried a mutation involving the replacement of Gly382 with Glu. This mutation inactivated the ability of PAT1 to complement the defective n-decane utilization of the disruptant. These results indicate that PAT1 encodes peroxisomal acetoacetyl-CoA thiolase and is essential for n-decane utilization in Y. lipolytica. PMID- 11401540 TI - The SH2 domain containing inositol 5-phosphatase SHIP2 controls phosphatidylinositol 3,4,5-trisphosphate levels in CHO-IR cells stimulated by insulin. AB - The lipid phosphatase SHIP2 (SH2 domain containing inositol 5-phosphatase 2) has recently been shown to be a potent negative regulator of insulin signaling and insulin sensitivity in vivo. We show here that SHIP2 is expressed in Chinese hamster ovary cells overexpressing the insulin receptor (CHO-IR cells) and tyrosine phosphorylated upon insulin stimulation. We show that SHIP2, which is recruited in anti-phosphotyrosine immunoprecipitates in insulin-stimulated cells, accounts for the insulin sensitivity or apparent increase in activity reported by Guilherme et al. (J. Biol. Chem. 271, 29533-29536, 1996). Overexpression of SHIP2 led to a decrease of the insulin-dependent PIP3 production as well as Akt/PKB activation and MAPK stimulation. PMID- 11401541 TI - Repression of AIM-1 kinase mRNA as part of a program of genes regulated by Mpl ligand. AB - Megakaryocytes give rise to platelets that are essential for thrombosis and hemostasis. During development, megakaryocytes undergo an endomitotic cell cycle by which they skip late anaphase and cytokinesis to yield high ploidy cells. This process is regulated by the c-Mpl receptor ligand. In the current study we used differential display PCR as well as degenerate cloning of kinases to identify part of the program of genes regulated during Mpl ligand-induced differentiation. Several of the induced genes were identified as encoding metabolic proteins as carnitine palmitolytransferase, while other altered genes were identified as encoding kinases. Of these, AIM-1 kinase mRNA was severely downregulated by Mpl ligand at the onset of polyploidy in megakaryocytes. This effect was not related to message stability, but rather to a change in transcriptional rate. These data point to the potential importance of the transcriptional regulation of the AIM-1 gene for promoting megakaryocyte polyploidization. PMID- 11401542 TI - Molecular structure of dihydroorotase: a paradigm for catalysis through the use of a binuclear metal center. AB - Dihydroorotase plays a key role in pyrimidine biosynthesis by catalyzing the reversible interconversion of carbamoyl aspartate to dihydroorotate. Here we describe the three-dimensional structure of dihydroorotase from Escherichia coli determined and refined to 1.7 A resolution. Each subunit of the homodimeric enzyme folds into a "TIM" barrel motif with eight strands of parallel beta-sheet flanked on the outer surface by alpha-helices. Unexpectedly, each subunit contains a binuclear zinc center with the metal ions separated by approximately 3.6 A. Lys 102, which is carboxylated, serves as a bridging ligand between the two cations. The more buried or alpha-metal ion in subunit I is surrounded by His 16, His 18, Lys 102, Asp 250, and a solvent molecule (most likely a hydroxide ion) in a trigonal bipyramidal arrangement. The beta-metal ion, which is closer to the solvent, is tetrahedrally ligated by Lys 102, His 139, His 177, and the bridging hydroxide. L-Dihydroorotate is observed bound to subunit I, with its carbonyl oxygen, O4, lying 2.9 A from the beta-metal ion. Important interactions for positioning dihydroorotate into the active site include a salt bridge with the guanidinium group of Arg 20 and various additional electrostatic interactions with both protein backbone and side chain atoms. Strikingly, in subunit II, carbamoyl L-aspartate is observed binding near the binuclear metal center with its carboxylate side chain ligating the two metals and thus displacing the bridging hydroxide ion. From the three-dimensional structures of the enzyme-bound substrate and product, it has been possible to propose a unique catalytic mechanism for dihydroorotase. In the direction of dihydroorotate hydrolysis, the bridging hydroxide attacks the re-face of dihydroorotate with general base assistance by Asp 250. The carbonyl group is polarized for nucleophilic attack by the bridging hydroxide through a direct interaction with the beta-metal ion. During the cyclization of carbamoyl aspartate, Asp 250 initiates the reaction by abstracting a proton from N3 of the substrate. The side chain carboxylate of carbamoyl aspartate is polarized through a direct electrostatic interaction with the binuclear metal center. The ensuing tetrahedral intermediate collapses with C O bond cleavage and expulsion of the hydroxide which then bridges the binuclear metal center. PMID- 11401543 TI - RNA-catalyzed amino acid activation. AB - We have selected RNAs that perform a new reaction that chemically activates amino acids, paralleling mixed phosphate anhydride synthesis by protein aminoacyl transfer RNA synthetases. Care with recovery of the unstable reaction product was apparently essential to this selection. The best characterized RNA, KK13, requires only Ca2+ for reaction and is optimally active at low pH with KM = 50 mM and kcat = 1.1 min(-1) for activation of leucine. In conjunction with previous RNA-catalyzed aminoacyl-RNA synthesis, peptide bond formation, and RNA-based coding, these amino acid-activating RNAs complete an experimental demonstration that the four fundamental reactions of protein biosynthesis can be RNA-mediated. The appearance of translation in an RNA world is therefore supported. PMID- 11401544 TI - A stem-loop of Tetrahymena telomerase RNA distant from the template potentiates RNA folding and telomerase activity. AB - The ribonucleoprotein enzyme telomerase adds telomeric repeats to the ends of linear chromosomes. The Tetrahymena telomerase reverse transcriptase (TERT) protein and the telomerase RNA can be reconstituted into an active complex in vitro in rabbit reticulocyte lysates. We have probed the structure of the telomerase RNA in the reconstituted complex with RNases T1 and V1. Upon TERT binding to the RNA, sites of both protection and enhancement of cleavage were observed, suggesting potential protein-binding sites and conformational changes in the RNA. Especially prominent was a large region of RNase V1 protection in stem-loop IV. A number of loop IV mutants still bound TERT but showed drastic decreases in the level of telomerase activity and the loss of protein-dependent folding of the pseudoknot region of the telomerase RNA. The telomerase activity defect and the misfolding of the pseudoknot were partially separable, leading to the proposal of two functions for stem-loop IV: to aid in the folding of the pseudoknot and to function more directly in the active site of telomerase. Thus an RNA element far from the template makes a major contribution to Tetrahymena telomerase enzyme activity. PMID- 11401545 TI - Cloning, overexpression, and purification of aminoglycoside antibiotic nucleotidyltransferase (2'')-Ia: conformational studies with bound substrates. AB - Aminoglycoside nucleotidyltransferase (2'')-Ia [ANT (2'')-Ia] was cloned from Pseudomonas aeruginosa and purified from overexpressing Escherichia coli BL21(DE3) cells. The first enzyme-bound conformation of an aminoglycoside antibiotic in the active site of an aminoglycoside nucleotidyltransferase was determined using the purified aminoglycoside nucleotidyltransferase (2' ')-Ia. The conformation of the aminoglycoside antibiotic isepamicin, a psuedo trisaccharide, bound to aminoglycoside nucleotidyltransferase (2' ')-Ia has been determined using NMR spectroscopy. Molecular modeling, employing experimentally determined interproton distances, resulted in two different enzyme-bound conformations (conformer 1 and conformer 2) of isepamicin. Conformer 1 was by far the major conformer defined by the following average glycosidic dihedral angles: PhiBC = -65.26 +/- 1.63 degrees and PsiBC = -54.76 +/- 4.64 degrees. Conformer 1 was further subdivided into one major (conformer 1a) and two minor components (conformers 1b and 1c) based on the comparison of glycosidic dihedral angles PhiAB and PsiAB. The arrangement of substrates in the enzyme.metal-ATP.isepamicin complex was determined on the basis of the measured effect of the paramagnetic substrate analogue Cr(H2O)4ATP on the relaxation rates of substrate protons which were used to determine relative distances of isepamicin protons to the Cr3+. Both conformers of isepamicin yielded arrangements that satisfied the NOE restraints and the observed paramagnetic effects of Cr(H2O)4ATP. It has been suggested that aminoglycosides use both electrostatic interactions and hydrogen bonds in binding to RNA and that the contacts made by the A and B rings to RNA are the most important for binding [Fourmy, D., Recht, M. I., Blanchard, S. C., and Puglisi, J. D. (1996) Science 274, 1367-1371]. Comparisons based on the determined conformations of enzyme-bound aminoglycoside antibiotics also suggested that interactions of rings A and B with enzymes may be the major determinant in aminoglycoside binding to enzymes [Serpersu, E. H., Cox, J. R., DiGiammarino, E. L., Mohler, M. L., Ekman, D. R., Akal-Strader, A., and Owston, M. (2000) Cell Biochem. Biophys. (in press)]. The conformation of isepamicin bound to the aminoglycoside nucleotidyltransferase (2' ')-Ia, determined in this work, lent further support to this theory. Furthermore, comparison of enzyme-bound conformations of isepamicin to the RNA-bound conformation of gentamycin C1a also showed remarkable similarities between the enzyme-bound and RNA-bound aminoglycoside antibiotic conformations. These studies should aid in the design of effective inhibitors possessing a broad range of aminoglycoside-modifying enzymes as targets. PMID- 11401546 TI - Human alpha spectrin II and the FANCA, FANCC, and FANCG proteins bind to DNA containing psoralen interstrand cross-links. AB - Repair of DNA interstrand cross-links is a complex process critical to which is the identification of sites of damage by specific proteins. We have recently identified the structural protein nonerythroid alpha spectrin (alphaSpIISigma) as a component of a nuclear protein complex in normal human cells which is involved in the repair of DNA interstrand cross-links and have shown that it forms a complex with the Fanconi anemia proteins FANCA, FANCC, and FANCG. Using DNA affinity chromatography, we now show that alphaSpIISigma, present in HeLa cell nuclei, specifically binds to DNA containing psoralen interstrand cross-links and that the FANCA, FANCC, and FANCG proteins are bound to this damaged DNA as well. That spectrin binds directly to the cross-linked DNA has been shown using purified bovine brain spectrin (alphaSpIISigma1/betaSpIISigma1)2. Binding of the Fanconi anemia (FA) proteins to the damaged DNA may be either direct or indirect via their association with alphaSpIISigma. These results demonstrate a role for alpha spectrin in the nucleus as well as a new function for this protein in the cell, an involvement in DNA repair. alphaSpIISigma may bind to cross-linked DNA and act as a scaffold to help in the recruitment of repair proteins to the site of damage and aid in their alignment and interaction with each other, thus enhancing the efficiency of the repair process. PMID- 11401547 TI - Inhibition of the aminopeptidase from Aeromonas proteolytica by L leucinephosphonic acid. Spectroscopic and crystallographic characterization of the transition state of peptide hydrolysis. AB - The nature of the interaction of the transition-state analogue inhibitor L leucinephosphonic acid (LPA) with the leucine aminopeptidase from Aeromonas proteolytica (AAP) was investigated. LPA was shown to be a competitive inhibitor at pH 8.0 with a K(i) of 6.6 microM. Electronic absorption spectra, recorded at pH 7.5 of [CoCo(AAP)], [CoZn(AAP)], and [ZnCo(AAP)] upon addition of LPA suggest that LPA interacts with both metal ions in the dinuclear active site. EPR studies on the Co(II)-substituted forms of AAP revealed that the environments of the Co(II) ions in both [CoZn(AAP)] and [ZnCo(AAP)] become highly asymmetric and constrained upon the addition of LPA and clearly indicate that LPA interacts with both metal ions. The X-ray crystal structure of AAP complexed with LPA was determined at 2.1 A resolution. The X-ray crystallographic data indicate that LPA interacts with both metal centers in the dinuclear active site of AAP and a single oxygen atom bridge is absent. Thus, LPA binds to the dinuclear active site of AAP as an eta-1,2-mu-phosphonate with one ligand to the second metal ion provided by the N-terminal amine. A structural comparison of the binding of phosphonate-containing transition-state analogues to the mono- and bimetallic peptidases provides insight into the requirement for the second metal ion in bridged bimetallic peptidases. On the basis of the results obtained from the spectroscopic and X-ray crystallographic data presented herein along with previously reported mechanistic data for AAP, a new catalytic mechanism for the hydrolysis reaction catalyzed by AAP is proposed. PMID- 11401548 TI - C-terminal region amino acid substitutions contribute to catalytic differences between murine class alpha glutathione transferases mGSTA1-1 and mGSTA2-2 toward anti-diol epoxide isomers of benzo[c]phenanthrene. AB - The molecular basis for catalytic differences between structurally closely related murine class alpha glutathione (GSH) transferases mGSTA1-1 and mGSTA2-2 in the GSH conjugation of anti-diol epoxide isomers of benzo[c]phenanthrene (anti B[c]PDE) was investigated. GSH conjugation of both (-)- and (+)-enantiomers of anti-B[c]PDE was observed in the presence of mGSTA1-1 (60 and 40% GSH conjugation, respectively), whereas mGSTA2-2 exhibited a preference for the (-) anti-isomer (>97%). In addition, the specific activity of mGSTA2-2 toward the (-) anti-B[c]PDE isomer was relatively higher than that of mGSTA1-1. The amino acid sequences of mGSTA1-1 and mGSTA2-2 differ at 10 positions that are distributed in three sections. Section I contains amino acid residues in positions 65 and 95; section II contains residues in positions 157, 162, and 169, and section III contains residues in positions 207, 213, 218, 221, and 222. Enzyme activity measurements with chimeras of mGSTA1-1 and mGSTA2-2 revealed that amino acid substitutions in section III account for their differential enantioselectivity and catalytic activity toward anti-B[c]PDE. Site-directed mutagenesis of amino acid residues in section III of mGSTA2-2 with corresponding residues of mGSTA1-1 followed by activity measurements of the wild type and mutated enzymes indicates that leucine 207 and phenylalanine 221 may be critical for the high catalytic activity of mGSTA2-2 toward (-)-anti-B[c]PDE. Molecular modeling studies demonstrated that the active site of mGSTA1-1 accommodates both enantiomers of anti-B[c]PDE, whereas the (-)-anti-isomer interacts more favorably with active site residues in mGSTA2-2. The results of this study clearly indicate that amino acid substitutions in the C-terminal region contribute to catalytic differences between mGSTA1-1 and mGSTA2-2 with respect to anti-B[c]PDE. PMID- 11401549 TI - Calcium-dependent binding of annexin 12 to phospholipid bilayers: stoichiometry and implications. AB - Annexins (ANXs) are a superfamily of proteins whose functional hallmark is Ca2+ dependent binding to anionic phospholipids. Their core domains are usually composed of a 4-fold repeat of a conserved amino acid sequence, with each repeat containing a type II Ca2+ binding site that is generally thought to mediate Ca2+ dependent binding to the membrane. We now report that ANX12 binding to phospholipid vesicles is highly cooperative with respect to Ca2+ concentration (Hill constant approximately 7), thereby suggesting that more than the four well characterized type II Ca2+ binding sites are involved in phospholipid binding. Two independent approaches, a novel 45Ca2+ copelleting assay and isothermal titration calorimetry, indicate a stoichiometry of approximately 12 mol of Ca2+/mol of ANX12 for binding to phospholipid vesicles. On the basis of the "low affinity" Ca2+-binding sites in a number of ANX X-ray crystal structures, we propose a model for ANX12 bilayer binding that involves three types of Ca2+ sites in each of the four repeats. In this model, there is a complementarity between the spacing of the ANX12 Ca2+ binding sites and the spacing of the phospholipid headgroups in bilayers. We tested the implications of the model by manipulating the physical state of vesicles composed of phospholipids with saturated acyl chains with temperature and measuring its influence on ANX12 binding. ANX12 bound to vesicles in a Ca2+-dependent manner when the vesicles were in the liquid crystal phase but not when the phospholipid was in the gel phase. Furthermore, ANX12 bound initially to fluid bilayers remained bound when cooled to 4 degrees C, a temperature that should induce the gel phase transition. Overall, these studies suggest that ANX12 is well suited to being a Ca2+ sensor for rapid all-or none intercellular membrane-related events. PMID- 11401550 TI - The 2.7 A crystal structure of the activated FERM domain of moesin: an analysis of structural changes on activation. AB - Moesin binds to a large range of proteins through its N terminal FERM (band 4.1, ezrin, radixin, moesin) domain. In full-length moesin isolated from cells, this binding is masked by binding to the C-terminal domain of moesin (C-ERMAD). Activation takes place by phosphorylation of Thr 558 in the C-ERMAD, which releases the C-ERMAD. A recently determined crystal structure of a noncovalent complex of the FERM and C-ERMAD domains showed for the first time that the structure of the FERM domain consists of three subdomains, each of which is similar to known structures. The structure reported here also contains a unique 47-residue helix pointing away from the FERM domain at the start of the alpha domain, in agreement with secondary structure predictions. Removal of the C-ERMAD does not result in a huge rearrangement of the FERM domain, but comparison with the activated radixin structure shows a consistent set of small changes. Not surprisingly, the exposed C-ERMAD binding area interacts in crystal contacts. More interestingly, a negatively charged peptide binds to the inositol site in a crystal contact and causes a greater conformational change in the structure than inositol. PMID- 11401551 TI - Interaction of yeast iso-1-cytochrome c with cytochrome c peroxidase investigated by [15N, 1H] heteronuclear NMR spectroscopy. AB - The interaction of yeast iso-1-cytochrome c with its physiological redox partner cytochrome c peroxidase has been investigated using heteronuclear NMR techniques. Chemical shift perturbations for both 15N and 1H nuclei arising from the interaction of isotopically enriched 15N cytochrome c with cytochrome c peroxidase have been observed. For the diamagnetic, ferrous cytochrome c, 34 amides are affected by binding, corresponding to residues at the front face of the protein and in agreement with the interface observed in the 1:1 crystal structure of the complex. In contrast, for the paramagnetic, ferric protein, 56 amides are affected, corresponding to residues both at the front and toward the rear of the protein. In addition, the chemical shift perturbations were larger for the ferric protein. Using experimentally observed pseudocontact shifts the magnetic susceptibility tensor of yeast iso-1-cytochrome c in both the free and bound forms has been calculated with HN nuclei as inputs. In contrast to an earlier study, the results indicate that there is no change in the geometry of the magnetic axes for cytochrome c upon binding to cytochrome c peroxidase. This leads us to conclude that the additional effects observed for the ferric protein arise either from a difference in binding mode or from the more flexible overall structure causing a transmittance effect upon binding. PMID- 11401552 TI - Charge translocation coupled to electron injection into oxidized cytochrome c oxidase from Paracoccus denitrificans. AB - Electrons were discretely injected into oxidized cytochrome c oxidase in liposomes by laser flash excitation of bound ruthenium [II] bispyridyl, and the membrane potential was recorded by time-resolved electrometry. Membrane potential is generated in a fast phase when an electron is transferred from the excited dye, via the CuA center, to heme a at a relative dielectric depth d inside the membrane [Zaslavsky, D., Kaulen, A. D., Smirnova, I. A., Vygodina, T., and Konstantinov, A. A. (1993) FEBS Lett. 336, 389-393]. Subsequently, membrane potential may develop further in a slower event, which is due to proton transfer into the enzyme from the opposite side of the membrane [Ruitenberg, M., Kannt, A., Bamberg, E., Ludwig, B., Michel, H., and Fendler, K. (2000) Proc. Natl. Acad. Sci. U.S.A. 97, 4632-4636]. Here, we confirm that injection of the first electron into the fully oxidized cytochrome c oxidase from Paracoccus denitrificans is associated with a fast electrogenic 11 micros phase, but there is no further electrogenic phase up to 100 milliseconds when special care is taken to ensure that only fully oxidized enzyme is present initially. A slower electrogenic 135 micros phase only becomes apparent and grows in amplitude upon increasing the number of light flashes. This occurs in parallel with a decrease in amplitude of the 11 micros phase and correlates with the number of enzyme molecules that are already reduced by one electron before the flash. The electrogenic 135 micros phase does not appear with increasing flash number in the K354M mutant enzyme, where electron and proton transfer into the binuclear center is delayed. We conclude that the 135 micros phase, and its associated proton uptake, take place on electron injection into enzyme molecules where the binuclear heme a3-CuB site is already reduced by one electron, and that it is accompanied by oxidation of heme a with a similar time constant. Reduction of heme a is not associated with electrogenic proton uptake into the enzyme, neither in the fully oxidized nor in the one-electron-reduced enzyme. The extent of the electrogenic 135 micrcos phase also rules out the possibility that reduction of the binuclear center by the second electron would be coupled to proton translocation in addition to the electrogenic uptake of a proton. PMID- 11401553 TI - Structural determinants for potent, selective dual site inhibition of human pp60c src by 4-anilinoquinazolines. AB - The kinetic mechanisms for the inhibition of pp60(c-src) tyrosine kinase (Src TK) by 4-anilinoquinazolines, an important class of chemicals as protein kinase inhibitors, were investigated. 4-Anilinoquinazolines with a bulky group at the 4' position of the anilino group were shown to be competitive with both ATP and peptide, whereas molecules lacking such a bulky group only displayed an inhibition pattern typical of those competitive with ATP and noncompetitive with peptide. Modifications of the substituents on the carbocyclic ring did not perturb the inhibition pattern although the affinities of these modified inhibitors for Src TK were affected. Structural modeling of Src TK with inhibitor and peptide substrate bound indicated a direct atomic conflict between the bulky 4-position group and the hydroxy of the peptide tyrosyl to which the gamma phosphate of ATP is transferred during the kinase reaction. This atomic conflict would likely prevent simultaneous binding of both inhibitor and peptide, consistent with the observed kinetic competitiveness of the inhibitor with peptide. The dual site inhibitors appeared to have both enhanced potency and selectivity for Src TK. One such inhibitor, 4-(4'-phenoxyanilino)-6,7 dimethoxyquinazoline, had a 15 nM potency against Src TK and was selective over receptor tyrosine kinases VEGFR2 by 88-fold and C-fms by 190-fold. PMID- 11401554 TI - Cyclization of backbone-substituted peptides catalyzed by the thioesterase domain from the tyrocidine nonribosomal peptide synthetase. AB - The excised C-terminal thioesterase (TE) domain from the multidomain tyrocidine nonribosomal peptide synthetase (NRPS) was recently shown to catalyze head-to tail cyclization of a decapeptide thioester to form the cyclic decapeptide antibiotic tyrocidine A [Trauger, J. W., Kohli, R. M., Mootz, H. D., Marahiel, M. A., and Walsh, C. T. (2000) Nature 407, 215-218]. The peptide thioester substrate was a mimic of the TE domain's natural, synthetase-bound substrate. We report here the synthesis of modified peptide thioester substrates in which parts of the peptide backbone are altered either by the replacement of three amino acid blocks with a flexible spacer or by replacement of individual amide bonds with ester bonds. Rates of TE domain catalyzed cyclization were determined for these substrates and compared with that of the wild-type substrate, revealing that some parts of the peptide backbone are important for cyclization, while other parts can be modified without significantly affecting the cyclization rate. We also report the synthesis of a modified substrate in which the N-terminal amino group of the wild-type substrate, which is the nucleophile in the cyclization reaction, is replaced with a hydroxyl group and show that this compound is cyclized by the TE domain to form a macrolactone at a rate comparable to that of the wild-type substrate. These results demonstrate that the TE domain from the tyrocidine NRPS can catalyze cyclization of depsipeptides and other backbone-substituted peptides and suggest that during the cyclization reaction the peptide substrate is preorganized for cyclization in the enzyme active site in part by intramolecular backbone hydrogen bonds analogous to those in the product tyrocidine A. PMID- 11401555 TI - Generality of peptide cyclization catalyzed by isolated thioesterase domains of nonribosomal peptide synthetases. AB - The C-terminal thioesterase (TE) domains from nonribosomal peptide synthetases (NRPSs) catalyze the final step in the biosynthesis of diverse biologically active molecules. In many systems, the thioesterase domain is involved in macrocyclization of a linear precursor presented as an acyl-S-enzyme intermediate. The excised thioesterase domain from the tyrocidine NRPS has been shown to catalyze the cyclization of a peptide thioester substrate which mimics its natural acyl-S-enzyme substrate. In this work we explore the generality of cyclization catalyzed by isolated TE domains. Using synthetic peptide thioester substrates from 6 to 14 residues in length, we show that the excised TE domain from the tyrocidine NRPS can be used to generate an array of sizes of cyclic peptides with comparable kinetic efficiency. We also studied the excised TE domains from the NRPSs which biosynthesize the symmetric cyclic decapeptide gramicidin S and the cyclic lipoheptapeptide surfactin A. Both TE domains exhibit expected cyclization activity: the TE domain from the gramicidin S NRPS catalyzes head-to-tail cyclization of a decapeptide thioester to form gramicidin S, and the TE domain from the surfactin NRPS catalyzes stereospecific cyclization to form a macrolactone analogue of surfactin. With an eye toward generating libraries of cyclic molecules by TE catalysis, we report the solid-phase synthesis and TE mediated cyclization of a small pool of linear peptide thioesters. These studies provide evidence for the general utility of TE catalysis as a means to synthesize a wide range of macrocyclic compounds. PMID- 11401556 TI - Thermodynamics of electron transfer in oxygenic photosynthetic reaction centers: a pulsed photoacoustic study of electron transfer in photosystem I reveals a similarity to bacterial reaction centers in both volume change and entropy. AB - The thermodynamic properties of electron transfer in biological systems are far less known in comparison with that of their kinetics. In this paper the enthalpy and entropy of electron transfer in the purified photosystem I trimer complexes from Synechocystis sp. PCC 6803 have been studied, using pulsed time-resolved photoacoustics on the 1 micros time scale. The volume contraction of reaction centers of photosystem I, which results directly from the light-induced charge separation forming P(700+F(A)/F(B-) from the excited-state P700*, is determined to be -26 +/- 2 A3. The enthalpy of the above electron-transfer reaction is found to be -0.39 +/- 0.1 eV. Photoacoustic estimation of the quantum yield of photochemistry in the purified photosystem I trimer complex showed it to be close to unity. Taking the free energy of the above reaction as the difference of their redox potentials in situ allows us to calculate an apparent entropy change (TDeltaS) of +0.35 +/- 0.1 eV. These values of DeltaV and TDeltaS are similar to those of bacterial reaction centers. The unexpected sign of entropy of electron transfer is tentatively assigned, as in the bacterial case, to the escape of counterions from the surface of the particles. The apparent entropy change of electron transfer in biological system is significant and cannot be neglected. PMID- 11401557 TI - Thermodynamics of electron transfer in oxygenic photosynthetic reaction centers: volume change, enthalpy, and entropy of electron-transfer reactions in manganese depleted photosystem II core complexes. AB - We have previously reported the thermodynamic data of electron transfer in photosystem I using pulsed time-resolved photoacoustics [Hou et al. (2001) Biochemistry 40, 7109-7116]. In the present work, using preparations of purified manganese-depleted photosystem II (PS II) core complexes from Synechocystis sp. PCC 6803, we have measured the DeltaV, DeltaH, and estimated TDeltaS of electron transfer on the time scale of 1 micros. At pH 6.0, the volume contraction of PS II was determined to be -9 +/- 1 A3. The thermal efficiency was found to be 52 +/ 5%, which corresponds to an enthalpy change of -0.9 +/- 0.1 eV for the formation of the state P680+Q(A-) from P680*. An unexpected volume expansion on pulse saturation of PS II was observed, which is reversible in the dark. At pH 9.0, the volume contraction, the thermal efficiency, and the enthalpy change were -3.4 +/- 0.5 A3, 37 +/- 7%, and -1.15 +/- 0.13 eV, respectively. The DeltaV of PS II, smaller than that of PS I and bacterial centers, is assigned to electrostriction and analyzed using the Drude-Nernst equation. To explain the small DeltaV for the formation of P680+Q(A-) or Y(Z*)Q(A-), we propose that fast proton transfer into a polar region is involved in this reaction. Taking the free energy of charge separation of PS II as the difference between the energy of the excited-state P680* and the difference in the redox potentials of the donor and acceptor, the apparent entropy change (TDeltaS) for charge separation of PS II is calculated to be negative, -0.1 +/- 0.1 eV at pH 6.0 (P680+Q(A-)) and -0.2 +/- 0.15 eV at pH 9.0 (Y(Z*)Q(A-)). The thermodynamic properties of electron transfer in PS II core reaction centers thus differ considerably from those of bacterial and PS I reaction centers, which have DeltaV of approximately -27 A3, DeltaH of approximately -0.4 eV, and TDeltaS of approximately +0.4 eV. PMID- 11401558 TI - Thermodynamics of electron transfer in oxygenic photosynthetic reaction centers: volume change, enthalpy, and entropy of electron-transfer reactions in the intact cells of the cyanobacterium Synechocystis PCC 6803. AB - The volume and enthalpy changes for charge transfer in the 0.1-10 micros time window in photosynthetic reaction centers of the intact cells of Synechocystis PCC 6803 were determined using pulsed, time-resolved photoacoustics. This required invention of a method to correct for the cell artifact at the temperature of maximum density of water caused by the heterogeneous system. Cells grown under either white or red light had different PS I/PS II molar ratios, approximately 3 and approximately 1.7, respectively, but invariable action spectra and effective antenna sizes of the photosystems. In both cultures, the photoacoustic measurements revealed that their thermodynamic parameters differed strongly in the spectral regions of predominant excitation of PS I (680 nm) and PS II (625 nm). On correcting for contribution of the two photosystems at these wavelengths, the volume change was determined to be -27 +/- 3 and -2 +/- 3 A3 for PS I and PS II, respectively. The energy storage on the approximately 1 micros time scale was estimated to be 80 +/- 15% and 45 +/- 10% per trap in PS I and PS II, respectively. These correspond to enthalpies of -0.33 +/- 0.2 and -1 +/- 0.2 eV for the assumed formation of ion radical pairs P700+F(AB-) and Y(Z*)P680Q(A-), respectively. Taking the free energy of the above reactions as the differences of their redox potentials in situ, apparent entropy changes were estimated to be +0.4 +/- 0.2 and -0.2 +/- 0.2 eV for PS I and PS II, respectively. These values are similar to that obtained in vitro for the purified reaction center complexes on the microsecond time scale [Hou et al. (2001) Biochemistry 40, 7109-7116, 7117 7125]. The constancy of these thermodynamic values over a 2-fold change of the ratio of PS I/PS II is support for this method of in vivo analysis. Our pulsed PA method can correct the "cell" or heterogeneous artifact and thus opens a new route for studying the thermodynamics of electron transfer in vivo. PMID- 11401559 TI - Identification of essential residues for catalysis of rat intestinal phospholipase B/lipase. AB - Intestinal brush border membrane-associated phospholipase B/lipase (PLB/LIP) consists of four tandem homologous domains (repeats 1 through 4) and a COOH terminal membrane binding domain, and repeat 2 is the catalytic domain that catalyzes phospholipase A2, lysophospholipase, and lipase activities. We examined the structural basis of the catalysis of PLB/LIP with this unique substrate specificity by site-directed mutagenesis of recombinant repeat 2 enzyme. Ser414 and Ser459 within the active serine-containing consensus sequence G-X-S-X-G in the best-established lipase family were dispensable for activity. In contrast, substitution of Ala for Ser404 almost completely inactivated the three lipolytic activities of PLB/LIP, even though the gross conformation was not altered as determined by CD spectroscopy. Notably, this Ser is located within the conserved G-D-S-L sequence on the NH2-terminal side in lipolytic enzymes of another group proposed recently. Furthermore, mutagenesis and CD spectroscopic analyses suggested that Asp518 and His659, lying within conserved short stretches in the latter group of lipolytic enzymes, were essential for activity. These three essential residues are conserved in the known PLB/LIP enzymes, suggesting that they form the catalytic triad in the active site. These results indicate that PLB/LIP represents a distinct class of the lipase family. PLB/LIP is the first mammalian member of that family. Repeat 2 is equipped with the triad, but not the other repeats, accounting for why only repeat 2 is the catalytic domain. Replacing Thr406 with Gly, matching the enzyme's sequence to the lipase consensus sequence exactly, led to a great decrease in secretion and accumulation of inactive enzyme in the cells, suggesting a role of Thr406 in the structural stability. PMID- 11401560 TI - Catalytic reaction profile for alcohol oxidation by galactose oxidase. AB - Galactose oxidase is a remarkable enzyme containing a metalloradical redox cofactor capable of oxidizing a variety of primary alcohols during enzyme turnover. Recent studies using 1-O-methyl alpha-D-galactopyranoside have revealed an unusually large kinetic isotope effect (KIE) for oxidation of the alpha deuterated alcohol (kH/kD = 22), demonstrating that cleavage of the 6,6' di[2H]hydroxymethylene C-H bond is fully rate-limiting for oxidation of the canonical substrate. This step is believed to involve hydrogen atom transfer to the tyrosyl phenoxyl in a radical redox mechanism for catalysis [Whittaker, M. M., Ballou, D. P., and Whittaker, J. W. (1998) Biochemistry 37, 8426-8436]. In the work presented here, the enzyme's unusually broad substrate specificity has allowed us to extend these investigations to a homologous series of benzyl alcohol derivatives, in which remote (meta or para) substituents are used to systematically perturb the properties of the hydroxyl group undergoing oxidation. Quantitative structure-activity relationship (QSAR) correlations over the steady state rate data reveal a shift in the character of the transition state for substrate oxidation over this series, reflected in a change in the magnitude of the observed KIE for these reactions. The observed KIE values have been shown to obey a log-linear correlation over the substituent parameter, Hammett sigma. For the relatively difficult to oxidize nitro derivative, the KIE is large (kH/kD = 12.3), implying rate-limiting C-H bond cleavage for the oxidation reaction. This contribution becomes less important for more easily oxidized substrates (e.g., methoxy derivatives) where a much smaller KIE is observed (kH/kD = 3.6). Conversely, the solvent deuterium KIE is vanishingly small for 4-nitrobenzyl alcohol, but becomes significant for the 4-methoxy derivative (kH2O/kD2O = 1.2). These experiments have allowed us to develop a reaction profile for substrate oxidation by galactose oxidase, consisting of three components (hydroxylic proton transfer, electron transfer, and hydrogen atom transfer) comprising a single-step proton-coupled electron transfer mechanism. Each component exhibits a distinct substituent and isotope sensitivity, allowing them to be identified kinetically. The proton transfer component is unique in being sensitive to the isotopic character of the solvent (H2O or D2O), while hydrogen atom transfer (C-H bond cleavage) is independent of solvent composition but is sensitive to substrate labeling. In contrast, electron transfer processes will in general be less sensitive to isotopic substitution. Our results support a mechanism in which initial proton abstraction from a coordinated substrate activates the alcohol toward inner sphere electron transfer to the Cu(II) metal center in an unfavorable redox equilibrium, forming an alkoxy radical which undergoes hydrogen atom abstraction by the tyrosine-cysteine phenoxyl free radical ligand to form the product aldehyde. PMID- 11401561 TI - Carbon monoxide adducts of KatG and KatG(S315T) as probes of the heme site and isoniazid binding. AB - KatG, the catalase peroxidase from Mycobacterium tuberculosis, is important in the activation of the antitubercular drug, isoniazid. About 50% of isoniazid resistant clinical isolates contain a mutation in KatG wherein the serine at position 315 is substituted with threonine, KatG(S315T). The heme pockets of KatG and KatG(S315T) and their interactions with isoniazid are probed using resonance Raman (rR) spectroscopy to characterize their ferrous CO complexes. Three vibrational modes, C-O and Fe-C stretching and Fe-CO bending, are assigned using 12CO and 13CO isotope shifts. Two conformers are observed for KatG-CO and KatG(S315T)-CO. Resonance Raman features assigned to form I are consistent with it having a neutral proximal histidine ligand and the Fe-C-O moiety hydrogen bonded to a distal residue. The nu(C-O) band for form I is sharp, consistent with a conformationally homogeneous Fe-CO unit. Form II also has a neutral proximal histidine ligand but is not hydrogen bonded. This appears to result in a conformationally disordered Fe-CO unit, as evidenced by a comparatively broad C-O stretching band. The 13CO-sensitive bands assigned to form II are predominant in the KatG(S315T)-CO rR spectrum. Isoniazid binding is apparent from the resonance Raman signatures of both WT KatG-CO and KatG(S315T)-CO. Moreover, isoniazid binding elicits an increase in the form I population of wild-type KatG-CO while having little, if any, effect on the already low population of form I of KatG(S315T)-CO. Since oxyKatG (compound III) also contains a low-spin diatomic ligand-heme adduct (heme-O2), it is reasonable to suggest that it too would exist as a mixture of conformers. Because the small form I population of KatG(S315T)-CO correlates with its inability to activate INH, we hypothesize that form I plays a role in INH activation. PMID- 11401562 TI - Preferential binding of human full-length XPA and the minimal DNA binding domain (XPA-MF122) with the mitomycin C-DNA interstrand cross-link. AB - Nucleotide excision repair (NER) is an important cellular mechanism that removes radiation-induced and chemically induced damage from DNA. The XPA protein is involved in the damage recognition step of NER and appears to function by binding damaged DNA and recruiting other proteins to the site. It may also play a role in subsequent steps of NER through interaction with other repair proteins. Interstrand cross-links are of particular interest, since these lesions involve both strands of duplex DNA and present special challenges to the repair machinery. Using 14 and 25 bp duplex oligonucleotides containing a defined, well characterized single mitomycin C (MMC)-DNA interstrand cross-link, we have shown through gel shift analysis that both XPA and a minimal DNA binding domain of XPA (XPA-MF122) preferentially bind to MMC-cross-linked DNA with a greater specificity and a higher affinity (>2-fold) than to the same undamaged DNA sequence. This preferential binding to MMC-cross-linked DNA occurs in the absence of other proteins from the NER complex. Differences in binding affinity and specificity were observed among the different protein-DNA combinations that were both protein and DNA specific. Defining XPA-MMC-DNA interactions may aid in elucidating the mechanism by which DNA cross-links and other forms of DNA damage are recognized and repaired by the NER machinery in eukaryotic cells. PMID- 11401563 TI - Recognition of 16S rRNA by ribosomal protein S4 from Bacillus stearothermophilus. AB - Protein S4 is essential for bacterial small ribosomal subunit assembly and recognizes the 5' domain (approximately 500 nt) of small subunit rRNA. This study characterizes the thermodynamics of forming the S4-5' domain rRNA complex from a thermophile, Bacillus stearothermophilus, and points out unexpected differences from the homologous Escherichia coli complex. Upon incubation of the protein and RNA at temperatures between 35 and 50 degrees C under ribosome reconstitution conditions [350 mM KCl, 8 mM MgCl2, and 30 mM Tris (pH 7.5)], a complex with an association constant of > or = 10(9) M(-1) was observed, more than an order of magnitude tighter than previously found for the homologous E. coli complex under similar conditions. This high-affinity complex was shown to be stoichiometric, in equilibrium, and formed at rates on the order of magnitude expected for diffusion controlled reactions ( approximately 10(7) M(-1) x s(-1)), though at low temperatures the complex became kinetically trapped. Heterologous binding experiments with E. coli S4 and 5' domain RNA suggest that it is the B. stearothermophilus S4, not the rRNA, that is activated by higher temperatures; the E. coli S4 is able to bind 5' domain rRNA equally well at 0 and 37 degrees C. Tight complex formation requires a low Mg ion concentration (1-2 mM) and is very sensitive to KCl concentration [- partial differential[log(K)]/partial differential(log[KCl]) = 9.3]. The protein has an unusually strong nonspecific binding affinity of 3-5 x 10(6) M(-1), detected as a binding of one or two additional proteins to the target 5' domain RNA or two to three proteins binding a noncognate 23S rRNA fragment of the approximately same size. This binding is not as sensitive to monovalent ion concentration [- partial differential[log(K)]/partial differential(log[KCl]) = 6.3] as specific binding and does not require Mg ion. These findings are consistent with S4 stabilizing a compact form of the rRNA 5' domain. PMID- 11401564 TI - The human interferon- and estrogen-regulated ISG20/HEM45 gene product degrades single-stranded RNA and DNA in vitro. AB - The human ISG20/HEM45 gene was identified independently on the basis of its increased level of expression in response to either interferon or estrogen hormone. Notably, the encoded protein is homologous with members of the 3' to 5' exonuclease superfamily that includes RNases T and D, and the proofreading domain of Escherichia coli DNA polymerase I. We provide here direct biochemical evidence that Isg20 acts as a 3' to 5' exonuclease in vitro. This protein displays a pH optimum of approximately 7.0, prefers Mn2+ as a metal cofactor, and degrades RNA at a rate that is approximately 35-fold higher than its rate for single-stranded DNA. Along with RNase L, Isg20 is the second known RNase regulated by interferon. Previous data showed that Isg20 is located in promyelocytic leukemia (PML) nuclear bodies, known sites of hormone-dependent RNA polymerase II transcription and oncogenic DNA viral transcription and replication. The combined data suggest a potential role for Isg20 in degrading viral RNAs as part of the interferon regulated antiviral response and/or cellular mRNAs as a regulatory component of interferon and estrogen signaling. PMID- 11401565 TI - Dynamics of translesion DNA synthesis catalyzed by the bacteriophage T4 exonuclease-deficient DNA polymerase. AB - The mechanism and dynamics of translesion DNA synthesis were evaluated using primer/templates containing a tetrahydrofuran moiety designed to mimic an abasic site. Steady-state kinetic analysis reveals that the T4 DNA polymerase preferentially incorporates dATP across from the abasic site with 100-fold higher efficiency than the other nucleoside triphosphates. Under steady-state conditions, the catalytic efficiency of dATP incorporation across from an abasic site is only 220-fold lower than that across from T. Surprisingly, misincorporation across from T is favored 4-6-fold versus replication across an abasic site, suggesting that the dynamics of the polymerization cycle are differentially affected by formation of aberrant base pairs as opposed to the lack of base-pairing capabilities afforded by the abasic site. Linear pre-steady state time courses were obtained for the incorporation of any dNTP across from an abasic site, indicating that chemistry or a step prior to chemistry is rate limiting for the polymerization cycle. Low elemental effects (<3) measured by substituting the alpha-thiotriphosphate analogues for dATP, dCTP, and dGTP indicate that chemistry is not solely rate-limiting. Single-turnover experiments yield kpol/Kd values that are essentially identical to kcat/Km values and provide further evidence that the conformational change preceding chemistry is rate limiting. Extension beyond an A:abasic mispair is approximately 20-fold and 100 fold faster than extension beyond a G:abasic mispair or C:abasic mispair, respectively. Extension from the G:abasic or A:abasic site mispair generates significant elemental effects (between 5 and 20) and suggests that chemistry is at least partially rate-limiting for extension beyond either mispair. PMID- 11401566 TI - An essential residue in the flexible peptide linking the two idiosynchratic domains of bacterial tyrosyl-tRNA synthetases. AB - Tyrosyl-tRNA synthetase (TyrRS) from Bacillus stearothermophilus comprises three sequential domains: an N-terminal catalytic domain, an alpha-helical domain with unknown function, and a C-terminal tRNA binding domain (residues 320-419). The properties of the polypeptide segment that links the alpha-helical and C-terminal domains, were analyzed by measuring the effects of sequence changes on the aminoacylation of tRNA(Tyr) with tyrosine. Mutations F323A (Phe323 into Ala), S324A, and G325A showed that the side chain of Phe323 was essential but not those of Ser324 and Gly325. Insertions of Gly residues between Leu322 and Phe323 and the point mutation L322P showed that the position and precise orientation of Phe323 relative to the alpha-helical domain were important. Insertions of Gly residues between Gly325 and Asp326 and deletion of residues 330-339 showed that the length and flexibility of the sequence downstream from Gly325 were unimportant but that this sequence could not be deleted. Mutations F323A, -L, -Y, and -W showed that the essential property of Phe323 was its aromaticity. The Phe323 side chain contributed to the stability of the initial complex between TyrRS and tRNA(Tyr) for 2.0 +/- 0.2 kcal x mol(-1) and to the stability of their transition state complex for 4.2 +/- 0.1 kcal x mol(-1), even though it is located far from the catalytic site. The results indicate that the disorder of the C-terminal domain in the crystals of TyrRS is due to the flexibility of the peptide that links it to the helical domain. They identified Phe323 as an essential residue for the recognition of tRNA(Tyr). PMID- 11401567 TI - Diverse roles of metal ions in acyl-transferase ribozymes. AB - The dependence on metal ions for catalysis is one of the hallmark characteristics of ribozymes. Yet despite this universal reliance, the functional role of divalent ions in promoting RNA catalysis is manifold. In this study we elucidate some different roles metal ions play as catalytic cofactors, by comparing two functionally co-evolved acyl-transferase ribozymes. Earlier studies performed on the in vitro selected acyl-transferase ribozyme, E18 [Suga, H., Cowan, J. A., and Szostak, J. W. (1998) Biochemistry 28, 10118-10125], revealed the requirement of a fully hydrated (outer-sphere) Mg2+ ion for catalytic activity. Interestingly, one class of acyl-transferase ribozymes isolated from the same RNA pool as E18 displays a unique metal dependency and is believed to be interacting with inner sphere coordinated Mg2+ ions. New results show that one of these inner-sphere coordinating ribozymes, HS01, assumes a cloverleaf secondary structure closely resembling E18, yet apparently facilitates a distinct catalytic mechanism. Furthermore, the nature of the RNA-metal interaction(s) in HS01 seems to be dictating a unique reaction mechanism that exhibits a titratable moiety at a near neutral pK(a). In light of the critical role metal ions play in biochemistry and the proper function of RNAs, these results compare two distinct manners by which metals serve to promote the catalysis of the same reaction. PMID- 11401568 TI - Interaction of fluorescence labeled single-stranded DNA with hexameric DNA helicase RepA: a photon and fluorescence correlation spectroscopy study. AB - Fluorescence correlation spectroscopy (FCS) was used to characterize the interaction of fluorescence labeled single-stranded DNA (ssDNA) with hexameric RepA DNA-helicase (hRepA) encoded by plasmid RSF1010. The apparent dissociation constants, Kd(app), for the equilibrium binding of 12mer, 30mer, and 45mer ssDNA 5'-labeled with BFL to hRepA dimer in the presence of 0.5 mM ATPgammaS at pH 5.8 and 25 degrees C were determined to be 0.58 +/- 0.12, 0.52 +/- 0.07, and 1.66 +/- 0.32 microM, respectively. Binding curves are compatible with one binding site for ssDNA present on hRepA dimer, with no indication of cooperativity. At pH 7.6 in the presence of ATPgammaS and at pH 5.8 in the absence of ATPgammaS, complex formation between ssDNA and hRepA was too weak for measuring complete binding curves by FCS. Under these conditions, the dissociation constant, Kd(app), is in the range between 10 and 250 microM. The kinetics of complex formation at pH 5.8 are faster than the time resolution (approximately 10-20 s) of FCS experiments under pseudo-first-order conditions, with respect to BFL-ssDNA. Photon correlation spectroscopy (PCS) experiments yielded, within the experimental error range, the same values for the apparent hydrodynamic radii, R(h), of hRepA dimer and its complex with ssDNA as determined by FCS (R(h) = 6.6 +/- 1 nm). hRepA starts to aggregate under acidic conditions ( W60 > W104. This order changes to W60 > W104 > W118 in the molten globule state. Remarkably, the solvent accessibility of W118 in the alpha-LA molten globule is lower than that in the native state. The dynamic properties of the three tryptophan residues were examined by time-resolved fluorescence anisotropy decay studies. The overall rotation of the molecule can be observed in both the native and molten globule states. In the molten globule state, there is an increase in the extent of local backbone fluctuations with respect to the native state. However, the fluctuation is not sufficient to result in complete motional averaging. The three tryptophan residues in the native and molten globule states have different degrees of motional freedom, reflecting the folding pattern and dynamic heterogeneity of these states. Taken together, these studies provide new insight into the structure and dynamics of the alpha-LA molten globule, which serves as a prototype for partially folded proteins. PMID- 11401571 TI - Slow motion in the CAA*TTG sequence of a DNA decamer duplex studied by NMR. AB - In DNA duplexes, pyrimidine-purine steps are believed to be flexible or conformationally unstable. Indeed, several DNA crystal structures exhibit a multitude of conformations for CpA*TpG steps. The question arises of whether this structural flexibility is accompanied by dynamical flexibility, i.e., a question pertaining to the energy barrier between conformations. Except for TpA steps, slow motions on the microsecond-to-millisecond time scale have not been detected in duplexes until now. In the present study, such slow motion was investigated by 1H, 13C, and 15N NMR relaxation measurements on a DNA decamer d(CATTTGCATC)*d(GATGCAAATG). The DNA decamer was enriched with 15% 13C and 98% 15N isotopes for each adenosine and guanosine residue. Three lines of evidence support the notion of slow motion in the CAA*TTG moiety. Analysis of (15)N relaxation showed that the order parameter, S2, of guanosine imino NH groups was about 0.8, similar to that of CH groups for this oligomer. The strong temperature dependence of guanosine NH S2 in the CAA*TTG sequence indicated the presence of a large-amplitude motion. Signals of adenosine H8 protons in the CAA*TTG sequence were broadened in 2D 1H NOESY spectra, which also suggested the existence of slow motion. As well as being smaller than for other adenine residues, the 1H T2 values exhibited a magnetic field strength dependence for all adenosine H8 signals in the ATTTG*CAAAT region, suggesting slow motions more pronounced at the first adenosine in the CAA*TTG sequence but extending over the CAAAT*ATTTG region. This phenomenon was further examined by the pulse field strength dependence of the 1H, 13C, and 15N T1rho values. 1H and 13C T1rho values showed a pulse field strength dependence, but 15N T1rho did not. Assuming a two-site exchange process, an exchange time constant of 20-300 micros was estimated for the first adenosine in the CAA sequence. The exact nature of this motion remains unknown. PMID- 11401572 TI - The A245K mutation exposes another stage of the bacterial L-lactate dehydrogenase reaction mechanism. AB - The A245K mutant of Bacillus stearothermophilus L-lactate dehydrogenase has been expressed in Escherichia coli and purified. A qualitative change in the reaction mechanism prior to the hydride transfer step in the reverse direction in the mutant is revealed. Both transient and steady state characteristics of the mutant are presented and show in contrast to the wild-type enzyme where a rearrangement of an enzyme-NADH-pyruvate complex is rate-limiting that in the mutant the rearrangement is much faster and hydride transfer is the first slow step. The steady state is limited by a new second slower conformation change involving an NAD+ complex. The mutation may provide a valuable framework for inhibitor and drug design research. PMID- 11401573 TI - Spectral studies of tert-butyl isothiocyanate-inactivated P450 2E1. AB - Inactivation of cytochrome P450 2E1 by tert-butyl isothiocyanate (tBITC) resulted in a loss in the spectrally detectable P450-reduced CO complex. The heme prosthetic group does not appear to become modified, since little loss of the heme was observed in the absolute spectra or the pyridine hemochrome spectra, or in the amount of heme recovered from HPLC analysis of the tBITC-inactivated samples. Prolonged incubations of the inactivated P450 2E1 with dithionite and CO resulted in a recovery of both the CO complex and the enzymatic activity. Inactivated samples that were first reduced with dithionite for 1 h prior to CO exposure recovered their CO spectrum to the same extent as samples not pretreated with dithionite, suggesting that the major defect was an inability of the inactivated sample to bind CO. Spectral binding studies with 4-methylpyrazole indicated that the inactivated P450 2E1 had an impaired ability to bind the substrate. Enzymatic activity could not be restored with iodosobenzene as the alternate oxidant. EPR analysis indicated that approximately 24% of the tBITC inactivated P450 2E1 was EPR-silent. Of the remaining tBITC-inactivated P450 2E1, approximately 45% exhibited an unusual low-spin EPR signal that was attributed to the displacement of a water molecule at the sixth position of the heme by a tBITC modification to the apoprotein. ESI-LC-MS analysis of the inactivated P450 2E1 showed an increase in the mass of the apoprotein of 115 Da. In combination, the data suggest that tBITC inactivated P450 2E1 by binding to a critical active site amino acid residue(s). This modified amino acid(s) presumably acts as the sixth ligand to the heme, thereby interfering with oxygen binding and substrate binding. PMID- 11401574 TI - Binding and oxidation of alkyl 4-nitrophenyl ethers by rabbit cytochrome P450 1A2: evidence for two binding sites. AB - Although most cytochrome P450 (P450) reactions demonstrate saturation kinetics that fit to the standard Michaelis-Menten equation, there are important exceptions where sigmoidal or nonhyperbolic behavior is observed and have been fit instead to kinetic models involving two binding sites. To assess these models, we demonstrate the consistency of a two binding site model to interpret both steady-state kinetics and binding events. Rates of 4-nitrophenol and formaldehyde production from the O-demethylation of 1-methoxy-4-nitrobenzene by P450 1A2 isolated from rabbit liver produced biphasic plots, when plotted against substrate concentration. Experiments confirmed the absence of the further oxidation of the products. Recombinant rabbit P450 1A2 yielded the same maximal velocity and more marked biphasicity. Overall, these steady-state data fit well to kinetic models involving two binding sites. Steady-state studies of substrates with bulkier O-ethyl or O-isopropoxy groups indicated decreased affinity for the second site. Based on binding studies, the affinity of P450 1A2 for these substrates increased 200-fold with the larger alkyl groups. To analyze the single binding site model, competition studies were conducted with 1,4 phenyldiisocyanide and the alkyl 4-nitrophenyl ethers. Although the observed dissociation constants and the competing titrant demonstrated a linear dependence, the affinity for the competing titrant depended on the presence of the other titrant, which violates the single binding site model. Alternatively, we applied a two binding site model to these data to obtain dissociation constants for the binary and ternary complexes. The agreement between the dissociation constants for the heterogeneous complexes supports the appropriateness of the two binding site model. This novel finding for P450 1A2 may be more common than originally perceived for P450s. PMID- 11401575 TI - Effects of substitution at serine 40 of tyrosine hydroxylase on catecholamine binding. AB - Phosphorylation of Ser40 in the regulatory domain of tyrosine hydroxylase activates the enzyme by increasing the rate of dissociation of inhibitory catecholamines [Ramsey, A. J., and Fitzpatrick, P. F. (1998) Biochemistry 37, 8980-8986]. To probe the structural basis for this effect and to ascertain the ability of other amino acids to functionally replace serine and serine phosphate, the effects of replacement of Ser40 with other amino acids were determined. Only minor changes in the Vmax value and the Km values for tyrosine and tetrahydropterin were seen upon replacement of Ser40 with alanine, valine, threonine, aspartate, or glutamate, in line with the minor effects of phosphorylation on steady-state kinetic parameters. More significant effects were seen on the binding of dopamine and dihydroxyphenylalanine. The affinity of the S40T enzyme for either catecholamine was very similar to that of the wild-type enzyme, while the S40E enzyme was similar to the phosphorylated enzyme. The S40D enzyme had an affinity for DOPA comparable to the phosphorylated enzyme but a higher affinity for dopamine than the latter. With both catecholamines, the S40V and S40A enzymes showed intermediate levels of activation. The results suggest that the serine hydroxyl contributes to the stabilization of the catecholamine inhibited enzyme. In addition, the S40E enzyme will be useful in further studies of the effects of multiple phosphorylation on tyrosine hydroxylase, while the alanine enzyme does not provide an accurate mimic of the unphosphorylated enzyme. PMID- 11401576 TI - Structural basis for thermostability in aporubredoxins from Pyrococcus furiosus and Clostridium pasteurianum. AB - The structures of apo- and holorubredoxins from Pyrococcus furiosus (PfRd) and Clostridium pasteurianum (CpRd) have been investigated and compared using residual dipolar couplings to probe the origin of thermostability. In the native, metal (Fe or Zn) containing form, both proteins can maintain native structure at very high temperatures (>70 degrees C) for extended periods of time. Significant changes in either structure or backbone dynamics between 25 and 70 degrees C are not apparent for either protein. A kinetic difference with respect to metal loss is observed as in previous studies, but the extreme stability of both proteins in the presence of metal makes thermodynamic differences difficult to monitor. In the absence of metal, however, a largely reversible thermal denaturation can be monitored, and a comparison of the two apoproteins can offer insights into the origin of stability. Below denaturation temperatures apo-PfRd is found to have a structure nearly identical to that of the native holo form, except immediately adjacent to the metal binding site. In contrast, apo-CpRd is found to have a structure distinctly different from that of its holo form at low temperatures. This structure is rapidly lost upon heating, unfolding at approximately 40 degrees C. A PfRd mutant with the hydrophobic core mutated to match that of CpRd shows no change in thermostability in the metal-free state. A metal-free chimera with residues 1-15 of CpRd and the remaining 38 residues of PfRd is severely destabilized and is unfolded at 25 degrees C. Hence, the hydrophobic core does not seem to be the key determinant of thermostability; instead, data point to the hydrogen bond network centered on the first 15 residues or the interaction of these 15 residues with other parts of the protein as a possible contributor to the thermostability. PMID- 11401577 TI - The reversible two-state unfolding of a monocot mannose-binding lectin from garlic bulbs reveals the dominant role of the dimeric interface in its stabilization. AB - Allium sativum agglutinin (ASAI) is a heterodimeric mannose-specific bulb lectin possessing two polypeptide chains of molecular mass 11.5 and 12.5 kDa. The thermal unfolding of ASAI, characterized by differential scanning calorimetry and circular dichroism, shows it to be highly reversible and can be defined as a two state process in which the folded dimer is converted directly to the unfolded monomers (A2 if 2U). Its conformational stability has been determined as a function of temperature, GdnCl concentration, and pH using a combination of thermal and isothermal GdnCl-induced unfolding monitored by DSC, far-UV CD, and fluorescence, respectively. Analyses of these data yielded the heat capacity change upon unfolding (DeltaC(p) and also the temperature dependence of the thermodynamic parameters, namely, DeltaG, DeltaH, and DeltaS. The fit of the stability curve to the modified Gibbs-Helmholtz equation provides an estimate of the thermodynamic parameters DeltaH(g), DeltaS(g), and DeltaC(p) as 174.1 kcal x mol(-1), 0.512 kcal x mol(-1) x K(-1), and 3.41 kcal x mol(-1) x K(-1), respectively, at T(g) = 339.4 K. Also, the free energy of unfolding, DeltaG(s), at its temperature of maximum stability (T(s) = 293 K) is 13.13 kcal x mol(-1). Unlike most oligomeric proteins studied so far, the lectin shows excellent agreement between the experimentally determined DeltaC(p) (3.2 +/- 0.28 kcal x mol(-1) x K(-1)) and those evaluated from a calculation of its accessible surface area. This in turn suggests that the protein attains a completely unfolded state irrespective of the method of denaturation. The absence of any folding intermediates suggests the quaternary interactions to be the major contributor to the conformational stability of the protein, which correlates well with its X-ray structure. The small DeltaC(p) for the unfolding of ASAI reflects a relatively small, buried hydrophobic core in the folded dimeric protein. PMID- 11401578 TI - Role of phylogenetically conserved amino acids in folding of Na,K-ATPase. AB - This paper focuses on the amino acid sequence 708-TGDGVNDSPALKK in pig kidney Na,K-ATPase as one of the best conserved among P-type ATPases. In Ca-ATPase this sequence forms a strand-loop-helix structure as part of a Rossman fold next to the phosphorylation site. Substitution of polar residues in the investigated sequence interfered with high-level accumulation of mutant protein. Mutant alpha1 subunit protein only accumulated in membranes from yeast cells grown at 15 degrees C whereas wild-type protein accumulated at both 15 and 35 degrees C. A systematic screen for the molecular mechanism behind lack of accumulation of mutant protein at 35 degrees C showed that transcription and translation were unaffected by the mutations. To demonstrate in vivo protein folding problems, an unfolded protein response reporter system was constructed in yeast. In this strain, only expression of mutant Na,K-ATPase alpha1-subunit caused induction of the unfolded protein response at 35 degrees C, indicating folding problems in the ER. Lowering the expression temperature to 15 degrees C prevented induction of the unfolded protein response after mutant protein expression, indicating correct folding at this temperature. At the permissive temperature mutant proteins were able to escape the endoplasmic reticulum quality control, reach the plasma membrane, and bind ouabain with high affinity. Since mutants in the 708 TGDGVNDSPALKK segment had a thermo inactivation profile identical to that of wild type, they were classified as temperature-sensitive synthesis mutants. The results indicate that this segment contributes side chains of importance for overall folding and maturation of Na,K-ATPase and all other P-type ATPases. PMID- 11401579 TI - Soret-excited Raman spectroscopy of the spinach cytochrome b6f complex. Structures of the b- and c-type hemes, chlorophyll a, and beta-carotene. AB - Soret-excited resonance Raman (RR) spectra of the spinach cytochrome b6f complex (cyt b6f) are reported for the oxidized, native, ascorbate-reduced, and dithionite-reduced forms. Using excitations at 441.6, 413.1, and 406.7 nm, RR contributions of chlorophyll a, beta-carotene, the c-type heme of cytochrome f, and the b-type hemes of cytochrome b6 of the b6f complex were identified and the data compared to those previously obtained for the Rhodospirillum rubrum bc1 complex [Le Moigne, C., Schoepp, B., Othman, S., Vermeglio, A., and Desbois, A. (1999) Biochemistry 38, 1066-1076]. RR bands arising from the b(6)f-associated chlorophyll a and beta-carotene pigments were found to be particularly intense in the spectra excited at 441.6 nm. The frequencies of the phorbin skeleton of chlorophyll a at 1606, 1552, and 1525 cm(-1) are typical of a Mg atom with a single axial ligand. Strong RR bands corresponding to stretching or deformation modes of beta-carotene were detected at 1137, 1157, 1191, 1216, and 1531 cm(-1) in the different forms of cyt b6f. This set of frequencies is assigned to an all trans configuration of the polyene chain. The redox titrations of the b(6)f complex allow the characterization of RR bands of the three hemes. The nu10, nu2, nu3, and nu8 modes of reduced cyt f are detected at 1619, 1591, 1492, and 356 cm( 1), respectively. From this set of frequencies, one can conclude that the particular histidine/amine heme coordination found in the truncated soluble domain of cyt f is a specific feature of the entire cyt f included in the b6f complex. The frequencies of the nu2, nu8, and nu10 marker modes are consistent with different conformations for the two b-type hemes of cyt b6f. One of these hemes is strongly distorted (nu2, nu8, and nu10 at 1581, 351, and 1610 cm(-1), respectively), while the other one is planar (1586, 345, and 1618 cm(-1), respectively). Largely different structures for the b-type hemes appear to be a common property for the bc1/b6f complexes. PMID- 11401580 TI - Sodium-dependent steps in the redox reactions of the Na+-motive NADH:quinone oxidoreductase from Vibrio harveyi. AB - The Na+-translocating NADH:ubiquinone oxidoreductase (Na+-NQR) from Vibrio harveyi was purified and studied by EPR and visible spectroscopy. Two EPR signals in the NADH-reduced enzyme were detected: one, a radical signal, and the other a line around g = 1.94, which is typical for a [2Fe-2S] cluster. An E(m) of -267 mV was found for the Fe-S cluster (n = 1), independent of sodium concentration. The spin concentration of the radical in the enzyme was approximately the same under a variety of redox conditions. The time course of Na+-NQR reduction by NADH indicated the presence of at least two different flavin species. Reduction of the first species (most likely, a FAD near the NADH dehydrogenase site) was very rapid in both the presence and absence of sodium. Reduction of the second flavin species (presumably, covalently bound FMN) was slower and strongly dependent on sodium concentration, with an apparent activation constant for Na+ of approximately 3.4 mM. This is very similar to the Km for Na+ in the steady-state quinone reductase reaction catalyzed by this enzyme. These data led us to conclude that the sodium-dependent step within the Na+-NQR is located between the noncovalently bound FAD and the covalently bound FMN. PMID- 11401581 TI - Requirements for osmosensing and osmotic activation of transporter ProP from Escherichia coli. AB - Transporter ProP of Escherichia coli, a solute-H+ symporter, can sense and respond to osmotic upshifts imposed on cells, on membrane vesicles, or on proteoliposomes that incorporate purified ProP-(His)6. In this study, proline uptake catalyzed by ProP was used as a measure of its osmotic activation, and the requirements for osmosensing were defined using the proteoliposome system. The initial rate of proline uptake increased with decreasing external pH and increasing DeltaPsi, lumen negative. Osmotic upshifts increased DeltaPsi by concentrating lumenal K+, but osmotic activation of ProP could be distinguished from this effect. Osmotic activation of ProP resulted from changes in Vmax, though osmotic shifts also increased the KM for proline. Osmotic activation could be described as a reversible, osmotic upshift-dependent transition linking (at least) two transporter protein conformations. No correlation was observed between ProP activation and the position of the anions of activating sodium salts within the Hofmeister series of solutes. Both the magnitude of the osmotic upshift required to activate ProP and the ProP activity attained were similar for membrane-impermeant osmolytes, including NaCl, glucose, and PEG 600. The membrane permeant osmolytes glycerol, urea, PEG 62, and PEG 106 failed to activate ProP. Two poly(ethylene glycol)s, PEG 150 and PEG 200, were membrane-permeant and did not cause liposome shrinkage, but they did partially activate ProP-(His)6. PMID- 11401582 TI - Regulatory properties of tropomyosin effects of length, isoform, and N-terminal sequence. AB - The regulatory properties of naturally occurring tropomyosins (Tms) of differing lengths have been examined. These Tms span from 4 to 7 actin subunits. Native proteins have been used to study the common 7 actin-spanning skeletal and smooth muscle variants and expressed recombinant proteins to study the shorter fibroblast 5a, 5b, yeast Tm1 and yeast Tm2 Tms (6, 6, 5, and 4 actin-spanning variants, respectively). The yTm2 has been overexpressed in Escherichia coli with N-terminal constructs equivalent to those previously used for yTm1 [Maytum, R., et al. (2000) Biochemistry 39, 11913]. The regulation of myosin subfragment 1 (S1) binding to actin by Tm has been assessed using a sensitive S1 binding titration. The equilibrium between closed and open (C to M states, KT = 0.1-0.14) was similar for all vertebrate Tms. Apart from skTm where the apparent cooperative unit size (n) is the same as the structural size (n = 7 actin sites), the other vertebrate Tms that were studied exhibited large n values (n = 12-14). The yeast Tms also exhibited large values of n (6-9) in comparison to their structural sizes (4-5). The determined value of KT depended on the N-terminal sequence (KT = 0.15-1). These results are compared with the effect of S1 upon Tm's affinity for actin. The yeast Tms have regulatory parameters similar to those of skTm, but unlike skTm, S1 has little effect upon their actin affinity. This shows that an actin state with a high affinity for S1 and Tm is not necessary for regulation, and the higher affinity of S1 for actin in the presence of vertebrate Tms is probably the result of a direct interaction of S1 with Tm. PMID- 11401583 TI - Heparin antagonists are potent inhibitors of mast cell tryptase. AB - Tryptase may be a key mediator in mast cell-mediated inflammatory reactions. When mast cells are activated, they release large amounts of these tetrameric trypsin like serine proteases. Tryptase is present in a macromolecular complex with heparin proteoglycan where the interaction with heparin is known to be essential for maintaining enzymatic activity. Recent investigations have shown that tryptase has potent proinflammatory activity, and inhibitors of tryptase have been shown to modulate allergic reactions in vivo. Many of the tryptase inhibitors investigated previously are directed against the active site. In the present study we have investigated an alternative approach for tryptase regulation. We show that the heparin antagonists Polybrene and protamine are potent inhibitors of both human lung tryptase and of recombinant mouse tryptase (mouse mast cell protease 6). Protamine inhibited tryptase in a competitive manner whereas Polybrene showed noncompetitive inhibition kinetics. Treatment of tetrameric, active tryptase with Polybrene caused dissociation into monomers, accompanied by complete loss of enzymatic activity. The present report thus suggests that heparin antagonists potentially may be used in treatment of mast cell-mediated diseases such as asthma. PMID- 11401586 TI - Influential ideas and experimental progress in schizophrenia genetics research. PMID- 11401588 TI - Surgery for temporal lobe epilepsy is regaining favor. PMID- 11401587 TI - Researchers optimistic about sea change in cancer treatment. PMID- 11401589 TI - Worldwide interest in global access to drugs. PMID- 11401594 TI - MMR immunization and autism. PMID- 11401596 TI - Thyroid disease and primary pulmonary hypertension. PMID- 11401598 TI - Causes of traumatic death during pregnancy. PMID- 11401603 TI - Ultrasound markers of fetal Down syndrome. PMID- 11401604 TI - Ultrasound markers of fetal Down syndrome. PMID- 11401602 TI - Ultrasound markers of fetal Down syndrome. PMID- 11401600 TI - Interobserver agreement about cervical cytologic and histologic diagnosis. PMID- 11401606 TI - Association between the T29-->C polymorphism in the transforming growth factor beta1 gene and breast cancer among elderly white women: The Study of Osteoporotic Fractures. AB - CONTEXT: Transgenic animal experiments suggest that increased expression of transforming growth factor beta1 (TGF-beta1) is protective against early tumor development, particularly in breast cancer. A T-->C (thymine to cytosine) transition in the 29th nucleotide in the coding sequence results in a leucine to proline substitution at the 10th amino acid and is associated with increased serum levels of TGF-beta1. OBJECTIVE: To determine whether an association exists between this TGF-beta1 polymorphism and breast cancer risk. DESIGN, SETTING, AND PARTICIPANTS: The Study of Osteoporotic Fractures, a prospective cohort study of white, community-dwelling women aged 65 years or older who were recruited at 4 US centers between 1986 and 1988. Three thousand seventy-five women who provided sufficient clinical information, buffy coat samples, and adequate consent for genotyping are included in this analysis. MAIN OUTCOME MEASURE: Breast cancer cases during a mean (SD) follow-up of 9.3 (1.9) years, verified by medical chart review and compared by genotype. RESULTS: Risk of breast cancer was similar in the 1124 women with the T/T genotype (56 cases; 5.4 per 1000 person-years) and the 1493 women with the T/C genotype (80 cases; 5.8 per 1000 person-years) but was significantly lower (P =.01) in the 458 women with the C/C genotype (10 cases; 2.3 per 1000 person-years). In analyses that adjusted for age, age at menarche, age at menopause, estrogen use, parity, body mass index, and bone mineral density, women with the C/C genotype had a significantly lower risk of developing breast cancer compared with women with the T/T or T/C genotype (hazard ratio [HR], 0.36; 95% confidence interval [CI], 0.17-0.75). There was no significant difference between the risk for women with the T/C genotype compared with women with the T/T genotype (adjusted HR, 1.04; 95% CI, 0.73-1.48). CONCLUSIONS: Our findings suggest that TGF-beta1 genotype is associated with risk of breast cancer in white women aged 65 years or older. Because the T allele is the common variant and confers an increased risk, it may be associated with a large proportion of breast cancer cases. PMID- 11401611 TI - Hormone replacement therapy and prevention of nonvertebral fractures: a meta analysis of randomized trials. AB - CONTEXT: Hormone replacement therapy (HRT) is widely considered to reduce fractures, but this belief is based on observational data; evidence from randomized trials is lacking. OBJECTIVE: To conduct a systematic review of all randomized trials of HRT that have reported or collected nonvertebral fracture data but that may not have focused on fracture prevention. DATA SOURCES: The MEDLINE, EMBASE, Science Citation Index, and Cochrane Controlled Trials Register databases were searched from 1997 through 2000 and a search was conducted of all recent systematic reviews to identify older studies. Authors were contacted to establish whether fracture data had been collected but not reported. Researchers in the field and pharmaceutical companies also were contacted to try to identify unpublished studies. STUDY SELECTION: Trials were included in which participants had been randomized to at least 12 months of therapy and data on nonvertebral fractures at any other site and due to any cause were available. Of 70 initially identified studies, 22 were included in the analysis. DATA EXTRACTION: Both investigators extracted data independently and appraised trial quality according to the Jadad scale, which assesses the methods of randomization, concealment allocation, and reporting of withdrawals and dropouts. Disagreements were resolved by discussion. DATA SYNTHESIS: There was an overall 27% reduction in nonvertebral fractures in a pooled analysis (reduction favoring HRT in relative risk [RR], 0.73; 95% confidence interval [CI], 0.56-0.94; P =.02). This effect was greater among women randomized to HRT who had a mean age younger than 60 years (RR, 0.67; 95% CI, 0.46-0.98; P =.03). Among women with a mean age of 60 years or older, there was a reduced effect (RR, 0.88; 95% CI, 0.71-1.08; P =.22). For hip and wrist fractures alone, the effectiveness of HRT appeared more marked (RR, 0.60; 95% CI, 0.40-0.91; P =.02), particularly for women younger than 60 years (RR, 0.45; 95% CI, 0.26-0.79; P =.005). CONCLUSIONS: Our meta-analysis of randomized controlled trials of HRT noted a statistically significant reduction in nonvertebral fractures. However, this effect may be attenuated in older women. PMID- 11401607 TI - Validation of clinical classification schemes for predicting stroke: results from the National Registry of Atrial Fibrillation. AB - CONTEXT: Patients who have atrial fibrillation (AF) have an increased risk of stroke, but their absolute rate of stroke depends on age and comorbid conditions. OBJECTIVE: To assess the predictive value of classification schemes that estimate stroke risk in patients with AF. DESIGN, SETTING, AND PATIENTS: Two existing classification schemes were combined into a new stroke-risk scheme, the CHADS( 2) index, and all 3 classification schemes were validated. The CHADS( 2) was formed by assigning 1 point each for the presence of congestive heart failure, hypertension, age 75 years or older, and diabetes mellitus and by assigning 2 points for history of stroke or transient ischemic attack. Data from peer review organizations representing 7 states were used to assemble a National Registry of AF (NRAF) consisting of 1733 Medicare beneficiaries aged 65 to 95 years who had nonrheumatic AF and were not prescribed warfarin at hospital discharge. MAIN OUTCOME MEASURE: Hospitalization for ischemic stroke, determined by Medicare claims data. RESULTS: During 2121 patient-years of follow-up, 94 patients were readmitted to the hospital for ischemic stroke (stroke rate, 4.4 per 100 patient years). As indicated by a c statistic greater than 0.5, the 2 existing classification schemes predicted stroke better than chance: c of 0.68 (95% confidence interval [CI], 0.65-0.71) for the scheme developed by the Atrial Fibrillation Investigators (AFI) and c of 0.74 (95% CI, 0.71-0.76) for the Stroke Prevention in Atrial Fibrillation (SPAF) III scheme. However, with a c statistic of 0.82 (95% CI, 0.80-0.84), the CHADS( 2) index was the most accurate predictor of stroke. The stroke rate per 100 patient-years without antithrombotic therapy increased by a factor of 1.5 (95% CI, 1.3-1.7) for each 1-point increase in the CHADS( 2) score: 1.9 (95% CI, 1.2-3.0) for a score of 0; 2.8 (95% CI, 2.0-3.8) for 1; 4.0 (95% CI, 3.1-5.1) for 2; 5.9 (95% CI, 4.6-7.3) for 3; 8.5 (95% CI, 6.3 11.1) for 4; 12.5 (95% CI, 8.2-17.5) for 5; and 18.2 (95% CI, 10.5-27.4) for 6. CONCLUSION: The 2 existing classification schemes and especially a new stroke risk index, CHADS( 2), can quantify risk of stroke for patients who have AF and may aid in selection of antithrombotic therapy. PMID- 11401610 TI - Association between apolipoprotein E epsilon4 and sleep-disordered breathing in adults. AB - CONTEXT: Apolipoprotein E epsilon4(ApoE epsilon4) is a well-known risk factor for Alzheimer disease and cardiovascular disease. Sleep-disordered breathing occurs in Alzheimer disease patients and increases risks for cardiovascular disease. Complex interactions among sleep, brain pathology, and cardiovascular disease may occur in ApoE epsilon4 carriers. OBJECTIVE: To study whether genetic variation at the level of ApoE is associated with sleep-disordered breathing or sleep abnormalities in the general population. DESIGN, SETTING, AND PARTICIPANTS: Ongoing longitudinal cohort study of sleep disorders at a US university beginning in 1989, providing a population-based probability sample of 791 middle-aged adults (mean [SD] age, 49 [8] years; range, 32-68 years). MAIN OUTCOME MEASURE: Nocturnal polysomnography to evaluate apnea-hypopnea index. RESULTS: The probability of moderate-to-severe sleep-disordered breathing (apnea-hypopnea index >/=15%) was significantly higher in participants with epsilon4, independent of age, sex, body mass index, and ethnicity (12.0% vs 7.0%; P =.003). Mean (SEM) apnea-hypopnea index was also significantly higher in participants with ApoE epsilon4 (6.5 [0.6] vs 4.8 [0.3]; P =.01). These effects increased with the number of ApoE epsilon4 alleles carried. CONCLUSIONS: A significant portion of sleep-disordered breathing is associated with ApoE epsilon4 in the general population. PMID- 11401608 TI - Improving quality improvement using achievable benchmarks for physician feedback: a randomized controlled trial. AB - CONTEXT: Performance feedback and benchmarking, common tools for health care improvement, are rarely studied in randomized trials. Achievable Benchmarks of Care (ABCs) are standards of excellence attained by top performers in a peer group and are easily and reproducibly calculated from existing performance data. OBJECTIVE: To evaluate the effectiveness of using achievable benchmarks to enhance typical physician performance feedback and improve care. DESIGN: Group randomized controlled trial conducted in December 1996, with follow-up through 1998. SETTING AND PARTICIPANTS: Seventy community physicians and 2978 fee-for service Medicare patients with diabetes mellitus who were part of the Ambulatory Care Quality Improvement Project in Alabama. INTERVENTION: Physicians were randomly assigned to receive a multimodal improvement intervention, including chart review and physician-specific feedback (comparison group; n = 35) or an identical intervention plus achievable benchmark feedback (experimental group; n = 35). MAIN OUTCOME MEASURE: Preintervention (1994-1995) to postintervention (1997-1998) changes in the proportion of patients receiving influenza vaccination; foot examination; and each of 3 blood tests measuring glucose control, cholesterol level, and triglyceride level, compared between the 2 groups. RESULTS: The proportion of patients who received influenza vaccine improved from 40% to 58% in the experimental group (P<.001) vs from 40% to 46% in the comparison group (P =.02). Odds ratios (ORs) for patients of achievable benchmark physicians vs comparison physicians who received appropriate care after the intervention, adjusted for preintervention care and nesting of patients within physicians, were 1.57 (95% confidence interval [CI], 1.26-1.96) for influenza vaccination, 1.33 (95% CI, 1.05-1.69) for foot examination, and 1.33 (95% CI, 1.04-1.69) for long-term glucose control measurement. For serum cholesterol and triglycerides, the achievable benchmark effect was statistically significant only after additional adjustment for physician characteristics (OR, 1.40 [95% CI, 1.08-1.82] and OR, 1.40 [95% CI, 1.09-1.79], respectively). CONCLUSION: Use of achievable benchmarks significantly enhances the effectiveness of physician performance feedback in the setting of a multimodal quality improvement intervention. PMID- 11401612 TI - Perspectives on care at the close of life. Psychological considerations, growth, and transcendence at the end of life: the art of the possible. AB - Patients with life-threatening illnesses face great psychological challenges and frequently experience emotional distress. Yet, the end of life also offers opportunities for personal growth and the deepening of relationships. When physical symptoms and suffering are controlled, it is easier to address patients' central concerns-about their families, about their own psychological integrity, and about finding meaning in their lives. Optimal end-of-life care requires a willingness to engage with the patient and family in addressing these distinct domains. In addition to supporting growth of patients and their caregivers, physicians need to recognize the impact of psychiatric disorders such as depression, anxiety, and delirium at the end of life and develop skills in diagnosing and treating these syndromes. Comments of a patient with pancreatic cancer, his son, and his physician help illuminate the potential opportunities presented when coping with life-threatening illness. Enhanced understanding of the common psychological concerns of patients with serious illness can improve not only the clinical care of the patient, but also the physician's sense of satisfaction and meaning in caring for the dying. PMID- 11401609 TI - End-of-life discussions and preferences among persons with HIV. AB - CONTEXT: Deficits in advance care planning leave many patients and their physicians unprepared for decisions about end-of-life care. Even though the prognosis has improved for many persons with human immunodeficiency virus (HIV) infection, a need for planning remains. OBJECTIVE: To evaluate prevalence of end of-life discussions, use of advance directives, and preferences concerning end-of life care and their relationship with patient demographics, clinical status, psychosocial variables, and practitioner characteristics among HIV-infected persons. DESIGN, SETTING, AND PATIENTS: Cross-sectional survey of a US probability sample of 2864, which represents 231 400 adults receiving care for HIV, conducted from January 1996 to April 1997. MAIN OUTCOME MEASURES: Communication with physician regarding end-of-life issues, completion of an advance directive, preference for aggressiveness of care, and willingness to tolerate future permanent adverse health states. RESULTS: A total of 1432 patients (50%) discussed some aspect of end-of-life care with their practitioner and 1088 (38%) completed an advance directive. Patients were more likely to complete an advance directive after a physician discussion (odds ratio [OR], 5.82; 95% confidence interval [CI], 4.50-7.52). Practitioners discussed end-of life care less with blacks (OR, 0.57; 95% CI, 0.39-0.83) and Latinos (OR, 0.74; 95% CI, 0.55-0.98) than with whites. Women (OR, 1.39; 95% CI, 1.05-1.84) and patients with children in the household (OR, 1.53; 95% CI, 1.12-2.10) communicated the most with practitioners about end-of-life issues. Patients infected with HIV via injection drug use (OR, 0.64; 95% CI, 0.45-0.89) and those with less education communicated the least with physicians about end-of-life issues. Less denial, greater trust in one's practitioner, and longer patient practitioner relationship were associated with more advance care planning. CONCLUSIONS: Half of all persons infected with HIV are at risk of making end-of life decisions without prior discussions with their health care practitioners. Blacks, Latinos, intravenous drug users, and less educated individuals need advance care planning interventions in clinical HIV programs. PMID- 11401613 TI - Perspectives on care at the close of life. Initiating end-of-life discussions with seriously ill patients. PMID- 11401615 TI - Postmenopausal hormone therapy for prevention of fractures: how good is the evidence? PMID- 11401619 TI - Issues in securing the community's sanction before making an intervention. AB - Achieving successful community change is a complex and daunting task. Indeed there is substantial concern today that American communities may be less capable in dealing with challenges than in many times during the past. The article reviews the community from a systems perspective and identifies the major properties that must be identified before any community intervention. This includes the initiation of both internal and external change. It concludes with a checklist of items that must be addressed to bring community change and a call for increasing community efficacy to deal with challenges. PMID- 11401614 TI - Genetic susceptibility to breast cancer: from the roll of the dice to the hand women were dealt. PMID- 11401620 TI - Ethics and community-based education: balancing respect for the community with professional preparation. AB - Community-based education and service learning are becoming increasingly common in health and human services education. As students enter the community, several ethical dilemmas arise regarding the university's interaction with the community. This article explores clinical, agency, and community placements in terms of the relationships they engender between the university and the community. The article then outlines some ethical obligations of universities and faculty members and ethical dilemmas that arise in different placements. Finally, a fundamental ethical framework that may guide universities and faculty members in planning community-based educational experiences is proposed. PMID- 11401621 TI - Ethical issues in intervention outcomes. AB - This article discusses selected ethical issues involved with the outcomes of interventions. An intervention is a process of intrusion into value systems based on a relationship of mutual trust and shared expectations between the intervener and the participant. Too often, interveners focus on the outcome without considering the mediating variables that can influence the outcome; therefore, intervention outcomes may fail or be short-lived. Many ethical issues with respect to how interventions are designed and implemented can affect their success or failure. PMID- 11401622 TI - The ethical dimensions of cultural competence in border health care settings. AB - Through thematic stories of patient and provider interactions on the U.S.-Mexico border, this article challenges the commonly understood definition of culture. It explores areas of concern related to cultural competency and medical ethics. Stories outline issues related to communication and comprehension, use of interpreters, gender and sexual orientation, traditional health care practices, socioeconomic status, age, health care settings, and involvement of community representatives. Policy recommendations address language, continuity of care, and health care professions education. PMID- 11401623 TI - Two major ethical issues in health education and promotion: assessing stage of change and cancer screening. AB - In community health education for behavioral change, valid assessment of stage of change is critical; in health promotion screening for cancer, health insurance coverage is also critical because some communities do not have the freedom to choose to change or to get their cancer treated. Proceeding without valid assessment of stage of change or insurance coverage can cause us harm. Harm comes to the patients when providers label them negatively or cause them anxiety if they cannot afford diagnosis and treatment beyond the screening. Not doing harm, (nonmaleficence) is our most compelling ethics principle. The authors analyze these two issues through the principles of autonomy, justice, and nonmaleficence in both their practice and that of their student with US-Mexico border communities and give recommendations. PMID- 11401625 TI - Ethical considerations in research with bereaved families. AB - This article describes a research project aimed at delivering an intervention to bereaved family members living in the community. The issues covered are access to subjects, recruitment and retention of subjects, and random assignment to control and experimental groups. PMID- 11401624 TI - Ethical issues in community-based cancer control: considerations in designing interventions. AB - Intervention studies are essential in the drive to reduce the burden of cancer in the United States. The means to accomplish primary and secondary cancer prevention is possible through health education focused on smoking, dietary changes, and the judicious application of screening technologies. The goal is to demonstrate these can work in the "real world" of the community workplace. The challenge of designing and conducting effective studies must include practical solutions to ethical as well as methodological issues. PMID- 11401626 TI - Big is significantly more beautiful? PMID- 11401627 TI - Optic neuritis and multiple sclerosis. AB - PURPOSE: Review of the association between optic neuritis (ON) and multiple sclerosis (MS). RESULTS: MS often presents as acute unilateral ON. While it is clear that many patients with ON suffer from a generalized disease of the central nervous system that will go on to clinically definite MS (CDMS), it is also clear that others do not. With more and more well-informed patients and the emerging pharmacotheraphy for MS, the distinction between those patients with ON who have MS and those who do not, has become more important than ever before. Recently, a large randomized clinical trial on patients with ON or other clinically isolated syndromes suggestive of MS and evidence of prior subclinical demyelination on magnetic resonance imaging of the brain, found that treatment with recombinant interferon-beta-1a is beneficial by reducing the development of CDMS. CONCLUSION: Ophthalmologists should refer their patients with acute ON to a neurologist for MS-directed investigations and decisions regarding early institution of disease modifying therapy. PMID- 11401628 TI - The development of myopia in Hong Kong children. AB - PURPOSE: The purpose of this two and one-half year follow-up study is to evaluate the relationship between early academic activities and the development of myopia. METHODS: 128 children randomly selected from 51 pre-schools completed an education and vision assessment study. At the end of the period, the children were 8.5 years old. Data were collected on developmental performance in the areas of cognition, language, social, academic and motor skills. The prevalence of myopia, mean refraction and change of refraction over this period were described in terms of the gender, geographical areas, type of pre-schools and type of primary schools. RESULTS: The prevalence of myopia shifted towards a higher proportion of myopia in all the above categories. The mean refraction at the initial visit was 0.5 D and became about -0.50 D at the follow-up visit. There was no difference in the changes of refraction in relation to the type of pre school or primary school. The change of refraction was also not related to the educational scores. However, there was a statistically significant difference in the academic scores, in particular the reading scores, between the activity and conventional primary schools. There was also gender difference in language scores between the girls and the boys of the activity schools. There was no difference in all scores between the myopes and non-myopes. CONCLUSION: The present study did not demonstrate a relationship between early academic activities and development of myopia. However, the higher reading scores and the slightly faster progression rate in the conventional schools may indicate an influence of the curriculum on the refractive development. A longer follow-up study may elucidate a clearer trend. PMID- 11401631 TI - Visual screening of Swedish children: an ophthalmological evaluation. AB - PURPOSE: This study describes the various ophthalmological conditions detected in the Swedish visual screening program for children. METHODS: The study was longitudinal and retrospective. All children (3126) born in 1982 in three Swedish municipalities have been followed from birth to ten years of age. Visual acuity was examined at the ages of 4, 5.5, 7 and 10 years. Before the age of 4, a gross examination of the eyes was performed. RESULTS: The prevalence of ametropia in the population was 7.7%, the prevalence of strabismus 3.1%, and the prevalence of organic lesions 0.6%. Seven children (0.2%) were visually handicapped (visual acuity or = 30 mmHg occurred in 39.5% of the eyes. After one week, IOP had returned to the preoperative level. After 4 months, IOP had further decreased to 16.1 +/- 3.8 mmHg (p = 0.0027). After a mean follow-up of 1-3.7 (mean 2.8) years, the average postoperative IOP was 15.1 +/- 2.9 mmHg, being significantly (p = 0.001) lower than the preoperative IOP with 86% of the patients having a mean of 1.6 drugs on average. The type of incision (scleral vs. corneal) did not affect the postoperative IOP level. Using the criteria of Bigger and Becker (1971) the long-term IOP control after PHACO-IOL surgery was improved or unchanged in 86% and worse in 14% of the preoperatively well-controlled OAG eyes. CONCLUSIONS: In OAG eyes PHACO IOL is associated with a significant decrease in IOP with less medication up to 1-3.7 (mean 2.8) years. PMID- 11401648 TI - Macular star associated with posterior hyaloid detachment. AB - PURPOSE: To report a patient with a Leber's idiopathic stellate neuroretinitis like lesion that was caused by vitreous traction accompanying a posterior hyaloid detachment. METHODS: We present a 49-year-old woman who showed segmental optic disc edema, peripapillary retinal detachment, and a macular star. RESULTS: An incomplete posterior hyaloid detachment was present, and the posterior hyaloid membrane was attached to the nasal margin of the optic disc. Fluorescein angiography revealed a bent retinal artery on the upper margin of the optic disc, and leakage of fluorescein was observed from this area. CONCLUSION: Vascular damage to the optic disc due to vitreous traction should also be considered as a mechanism for the ophthalmoscopic appearance of the fundus when lesions suggestive of Leber's idiopathic stellate neuroretinitis are present in an adult. PMID- 11401652 TI - Alzheimer's peptide: a possible link between glaucoma, exfoliation syndrome and Alzheimer's disease. PMID- 11401650 TI - Uveal melanomas with optic nerve extension: report of two cases diagnosed by transvitreal biopsy, one of them with a multicentric tumour. AB - PURPOSE: To report two cases of invasive uveal melanomas, one of which showed multifocality. METHODS: Clinical examination, ultrasonography, colour Doppler analysis, cytological and histopathological evaluations. RESULTS: Transvitreal biopsy (case 1) or fine needle aspiration biopsy (FNAB) (case 2) revealed malignant melanomas in both patients. Light microscopy and immunohistochemical examinations substantiated the diagnosis of mixed cell type melanomas. In addition, one patient had a multifocal melanoma with papilloedema and colour Doppler findings suggestive of optic disc involvement. CONCLUSION: Transvitreal biopsy for histology or cytology is a reliable procedure to obtain an accurate diagnosis without delay of a lesion adjacent to the optic nerve head. In our two cases the biopsy findings led to enucleation. PMID- 11401651 TI - Removal of a large non-functional bleb and reconstruction with free conjunctival autograft after trabeculectomy. PMID- 11401653 TI - Antenatal sonographic diagnosis of dacryocystocele. PMID- 11401654 TI - A study about late age onset depression. PMID- 11401655 TI - The genetics of autism. AB - OBJECTIVE: To review systematically the empirical evidence for the involvement of genetic risk factors in infantile autism. METHOD: We aimed at including all relevant papers written in English. We conducted a Medline search in September 2000. In addition we searched the reference lists of related papers. RESULTS: A relatively small number of reports including family and twin studies, comorbidity, cytogenetic and molecular genetic studies were reviewed. CONCLUSION: As well family, twin, cytogenetic and molecular genetic studies supported the importance of genetic risk factors in infantile autism. In most individual cases probably at least a few gene variants simultaneously determine the genetic risk. Presently the most interesting chromosome regions concerning the aetiology of autism are chromosomes 7q31-35, 15q11-13 and 16p13.3 which have been suggested by different lines of genetic research. PMID- 11401656 TI - Screening for autism spectrum disorders in adult psychiatric out-patients: a preliminary report. AB - OBJECTIVE: To estimate the prevalence of autism spectrum disorders (ASD) among adult psychiatric out-patients; to evaluate the efficacy of a new brief screening questionnaire (ASDASQ). METHOD: 1323 adult psychiatric out-patients were screened by staff. Analysis of psychiatric records of patients (n = 66) scoring high on the ASDASQ yielded 31 patients with a suspected ASD. Twenty-two of these patients were clinically examined. Three psychometric aspects of the questionnaire were studied. RESULTS: Seventeen patients were found by clinical examination to have an ASD. Since two patients scoring low on the ASDASQ were known to have an ASD, at least 19 patients in this population (1.4%) had a definite ASD. Seventeen of the ASD patients had been previously diagnosed with other psychiatric disorders, most frequently schizophrenia (n = 5). Of patients attending a treatment centre for severe psychiatric disabilities (n = 499), 3.2% had an ASD. The ASDASQ showed good reliability across and within raters. Internal consistency was excellent. CONCLUSION: Adult psychiatric patients sometimes have undiagnosed autism spectrum disorders. The ASDASQ can be useful for screening. PMID- 11401657 TI - Citalopram versus nortriptyline in late-life depression: a 12-week randomized single-blind study. AB - OBJECTIVE: The aim of this single-blind study was to examine the efficacy and tolerability of citalopram compared to nortriptyline in moderate to severe major depressive patients aged 60 years or over. METHOD: In- and out-patients (N=58) with unipolar major depression were randomized to 12-week flexible dose treatment with nortriptyline or citalopram. RESULTS: No significant differences between the number of drop-outs in either group were observed, but the autonomic side-effects were significantly higher for nortriptyline than for citalopram. A significantly higher remission rate to nortriptyline than to citalopram was demonstrated, particularly if severe patients (endogenous or psychotic patients) were assessed. CONCLUSION: The remission rate to a therapeutic plasma level of nortriptyline appears to be higher than the remission rate to a standard dose of citalopram in a group of elderly major depressed patients, especially those with endogenous or psychotic features. On the other hand, citalopram appears to be better tolerated. PMID- 11401658 TI - The Parma High School Epidemiological Survey: obsessive-compulsive symptoms. AB - OBJECTIVE: This study was undertaken to assess the prevalence of obsessive compulsive symptoms in a population of Italian adolescents. METHOD: A sample of 2877 high-school students, 1463 males (51%) and 1414 females (49%), aged 16-21 years were assessed with the Leyton Obsessional Inventory--Child Version (LOI CV). Two groups of subjects were defined as 'positive' on the screen: the high interference (HI) (i.e. all subjects who scored 25 or more in the interference score), and the supernormals (Sn). RESULTS: Females scored higher than males both on yes and interference scores. One hundred and nineteen (4.1%) and 87 (3.0%) constitute, respectively, the HI and the Sn groups. The most interfering symptoms were obsessions linked to dirt phobia, rumination and nail biting. CONCLUSION: The prevalence estimates of OCD symptoms confirmed the recent data of the literature that adolescent OCD symptoms are more frequent than was believed previously. Females showed more symptoms and more interference on personal functioning than males. PMID- 11401660 TI - Maastricht Assessment of Coping Strategies (MACS-I): a brief instrument to assess coping with psychotic symptoms. AB - OBJECTIVE: To examine the reliability of a brief instrument to assess coping with symptoms by patients with psychotic illness. METHOD: A semi-structured interview (MACS) was developed to assess the amounts of distress, control and coping in relation to psychotic symptoms. Two raters interviewed 23 symptomatic but stable patients with a diagnosis of chronic schizophrenia on two separate occasions. Case managers were also interviewed. RESULTS: Both the number of coping strategies used for different groups of symptoms, and the amount of coping used in different domains of coping strategies could be assessed reliably between interviewers (intraclass correlation coefficients range: 0.90-0.97) and between interview sessions (ICCs range: 0.75-0.80). Reliability with case managers, however, was low. CONCLUSION: The MACS may be a reliable and useful instrument to assess coping in relation to subjective experience of distress by and control over psychotic symptoms. PMID- 11401662 TI - A structured interview guide increases Brief Psychiatric Rating Scale reliability in raters with low clinical experience. AB - OBJECTIVE: To assess the beneficial impact of a structured interview on the reliability of BPRS ratings in raters with low clinical experience. METHOD: Each patient was rated once a week in two separate interviews, conducted on the same day. The first interview was conducted by a rater with low clinical experience (recruited from a group of five residents in psychiatry and one clinical psychologist in training). All second interviews were conducted by the same highly experienced psychiatrist. RESULTS: The number of items with full agreement between observers increased with the use of SIG. The value of intraclass correlation coefficients for individual items and the total score also increased, approaching reported studies with experienced raters. CONCLUSION: These results suggest that the use of SIG reduces variability of information gathering in reliability testing of BPRS with less experienced raters. PMID- 11401663 TI - OCD and transvestism: is there a relationship? AB - OBJECTIVE: There have been reports of obsessive-compulsive disorder (OCD) patients with comorbid paraphilias. In this paper, two cases of comorbidity between OCD and transvestism are reported with the aim of discussing possible explanations for this association. METHOD: Longitudinal case studies of two patients with OCD and tranvestism according to the DSM-IV were assessed using semi-structured interviews. RESULTS: Both patients presented with OCD and comorbid transvestism with different clinical features and treatment response. The case whose cross-dressing was more egodystonic responded better to treatment. CONCLUSION: It is possible that some cases of transvestism are OCD-related while others are more related to a gender identity disorder. A dimensional approach focusing on common phenomenological and neurobiological substrates is suggested as particularly useful for clinical management and future research of both disorders. PMID- 11401661 TI - MACS-II: does coping enhance subjective control over psychotic symptoms? AB - OBJECTIVE: Associations between subjective experience of control and the use of self-initiated coping strategies were examined in patients with psychotic symptoms. METHOD: Twenty-three patients were interviewed to assess (i) the subjective experience of distress with and control over symptoms and (ii) the coping strategies used. RESULTS: There was a positive association between coping type and control (OR = 1.07, 95% CI: 1.03-1.12). Active coping strategies had the strongest association with experience of control (active problem-solving: OR = 1.41 95% CI: 1.18-1.68; active problem-avoiding: OR = 1.45 95% CI: 1.18-1.68). Symptomatic coping was negatively associated with control (OR = 0.40, 95% CI: 0.30-0.55), but was the most frequently used strategy. Depressive symptoms were associated with the highest number of coping strategies. CONCLUSION: Coping strategies differ in the degree and direction of associated subjective control, and symptoms differ in the degree with which coping is mobilized. Assessment of coping strategies may be useful as a prelude to psychological therapy. PMID- 11401659 TI - A prospective study of 86 new patients with social anxiety disorder. AB - OBJECTIVE: To determine the prognosis in 86 new patients with social anxiety disorder. METHOD: Untreated subjects with social anxiety were recruited by advertising in Stockholm, randomized to 3 months treatment with paroxetine or placebo, and then offered continued specialist care. Metabolizing capacity was determined by genotyping CYP2D6 in the subjects on paroxetine. After a mean 32 months all were contacted for a personal interview. RESULTS: Of the 92 evaluable subjects, 86 (93%) were interviewed. A favourable prognosis was seen in the subjects randomized to paroxetine who chose to continue with serotonergic medication. The least favourable prognosis was in those given placebo who chose not to be treated after the trial. Twenty-four subjects were still symptomatic and dysfunctional and had not sought treatment. Drug-induced adverse effects caused treatment termination in six subjects, one of whom had a poor metabolizing genotype. CONCLUSION: Due to their condition, some subjects with social anxiety refrain from effective treatments. The efficacy of serotonergic medication was maintained and augmented after a mean period of 32 months. PMID- 11401665 TI - Sea-urchin granuloma: histologic profile. A pathologic study of 50 biopsies. AB - BACKGROUND: Sea-urchin granuloma is a chronic granulomatous skin lesion caused by injury with sea-urchin spines. Frequently these lesions occur on the hands and develop several months after the initial injury. Classified as an allergic foreign-body reaction, their most common histological pattern resembles sarcoid. The purpose of this study was to evaluate the light microscopic features of biopsies from lesions clinically diagnosed of sea-urchin granolomas. METHODS: We retrospectively reviewed 50 biopsy specimens corresponding to 35 patients with sea-urchin granulomas. These lesions were caused by injuries with the spines of the sea-urchin Paracentrotus lividus. Data were collected between 1990 and 1999 from patients in the seashore of Galicia (NW Atlantic coast, Spain). RESULTS: The cohort consisted of 35 patients (31 males, 4 females), with a median age of 35 years (range 14-60 years). The median duration of the disease was 7.5 months (range 2-60 months). We identified different histopathologic patterns. A granulomatous reaction was observed in 39 biopsies (78%). In 70% corresponding to 35 biopsies this granulomatous reaction was predominant. Foreign-body, sarcoidal, tuberculoid, necrobiotic and suppurative granulomas were identified. The remaining 15 biopsies (30%) showed a predominant inflammatory reaction with features of non-specific chronic inflammation or suppurative dermatitis. A panel of histopathologic features, including epidermal and dermal changes were evaluated. Presence of focal necrosis and microabscesses were common findings. In 50% of our specimens we found umbilication and/or perforation. Additional features included the presence of inclusion epidermoid cysts in four cases and squamous syringometaplasia in one case. CONCLUSIONS: Our observations suggest that sea-urchin granuloma span a wide morphologic spectrum. A granulomatous inflammatory reaction was predominant, with the foreign body and sarcoidal types the most frequent patterns. Other histopathologic patterns with non granulomatous inflammation can be noted. Some features, such as the frequency of perforation and the presence of necrobiotic granulomas have not previously been recognized in the literature. PMID- 11401666 TI - S100A6 expression in fibrohistiocytic lesions. AB - BACKGROUND: S100A6, an S100 calcium-binding protein, has been found in a variety of cutaneous and extracutaneous lesions including: melanocytic nevi, melanoma, some salivary gland and epithelial tumors, and malignant fibrous histiocytoma (MFH). Dermal dendrocytes (DD) in the papillary dermis of skin also express S100A6 protein. We evaluated a variety of cutaneous fibrohistiocytic lesions to determine if the immunophenotype of S100A6 positivity can be expanded to include some or all of these lesions. METHODS: Formalin-fixed, paraffin-embedded tissues from fibrous papules (FP, 20), dermatofibromas (DF, 20), dermatofibrosarcoma protuberans (DFSP, 5), atypical fibroxanthomas (AFX, 5), oral fibromas (3), digital fibroma (1), and dermatomyofibroma (1) were evaluated with antibodies to S100A6, S100B, factor XIIIa, and MAC387 using a one-hour capillary action-based immunohistochemical procedure. RESULTS: DD in 20/20 FP, 19/20 DF, and 4/4 fibromas stained positively with anti-S100A6 in a pattern similar to anti-factor XIIIa. No DFSP cases stained with anti-S100A6. Anti-S100A6 showed superior staining to anti-factor XIIIa in 4/5 AFX cases. CONCLUSIONS: The immunophenotypes of some fibrohistiocytic lesions can be expanded to include S100A6 protein. With the exception of AFX, the use of anti-S100A6 does not appear to offer added benefit over anti-factor XIIIa in the differential diagnosis of fibrohistiocytic lesions. PMID- 11401667 TI - Lupus profundus, indeterminate lymphocytic lobular panniculitis and subcutaneous T-cell lymphoma: a spectrum of subcuticular T-cell lymphoid dyscrasia. AB - INTRODUCTION: The diagnosis and classification of lymphocytic lobular panniculitis (LLP) has historically proven to be a difficult challenge. We encountered 32 cases of primary LLP which could be categorized as: 1) lupus erythematosus profundus (LEP) (19 patients); 2) an indeterminate group termed indeterminate lymphocytic lobular panniculitis (ILLP) (6 patients); and 3) subcutaneous T-cell lymphoma (SCTCL) (7 patients). OBJECTIVE: We attempted to better define the subtypes of LLP by morphologic, phenotypic and genotypic features and to correlate those features to clinical presentation and outcome. METHOD: Skin biopsy material was studied by conventional light microscopy, through immunophenotyping performed on sections from paraffin-embedded, formalin fixed tissue and in some cases on sections of tissue frozen after receipt in physiological (Michel's) medium, and by polymerase chain reaction single-stranded conformational polymorphism analysis to assess for clonality of T-lymphocytes. Clinical features were correlated to histologic, phenotypic, and genotypic analyses. RESULTS: Patients with LEP had a prior diagnosis of LE or overlying skin changes which light microscopically were characteristic of LE. Patients with ILLP had no concurrent or prior history of LE, no systemic symptoms or cytopenias, and a clinical course not suggestive of lymphoma. Cases of SCTCL showed hemophagocytic syndrome and/or lesional progression with demise attributable to the disease. Lesions in all groups showed proximal extremity predilection. Females predominated in the LEP group. The average age of onset was 38, 40 and 55 years in the LEP, ILLP and SCTCL groups, respectively. Cytopenia was seen in 4 LEP patients; 1 also developed fever. In LEP and ILLP, lesions resolved with hydroxychloroquine and/or steroid therapy, with recurrences following cessation of therapy. In the SCTCL group 4 developed hemophagocytic syndrome, 4 died within 2 years of diagnosis, and 3 went into remission following chemotherapy. The LEP and SCTCL groups manifested histological similarities: dense perieccrine and lobular lymphocytic infiltration, lymphoid atypia, histiocytes with ingested debris, eosinophilic necrosis of the fat lobule and thrombosis. The atypical lymphocytes although pleomorphic did not have a cerebriform morphology. The infiltrate in ILLP had a similar cytomorphology and distribution with variable angioinvasion which in all save one case was of lesser intensity and was not associated with significant fat necrosis or vasculitis. Germinal centers, dermal/subcuticular mucin deposition and an atrophying interface dermatitis with hyperkeratosis and follicular plugging were largely confined to the LEP group. Erythrophagocytosis, characteristic of SCTCL, usually indicated a supervening subcuticular lymphoid dyscrasia when encountered in ILLP and LEP. SCTCL showed a selective loss of CD5 expression with or without diminution in CD7 and monoclonal CD3 expression. Of 4 cases studied, 3 showed a CD8 dominant infiltrate while 2 others exhibited CD56 and CD30 positivity, respectively. All cases of SCTCL with amplifiable DNA showed T-cell clonality. Similar molecular and phenotypic features indicative of subcuticular lymphoid dyscrasia were encountered in cases of LEP and ILLP including a reduction in CD5, CD7, and/or monoclonal CD3 expression, a preponderance of CD8 lymphocytes within the subcutaneous fat and T-cell clonality. These cases showed lymphoid atypia with variable erythrophagocytosis. Cases of phenotypically abnormal and/or clonal LEP showed one or more of local destruction, lesional size progression, fever, and cytopenias, but lesions responded to hydroxychloroquine and/or prednisone therapy and death attributable to panniculitis could not be documented. Cases that were phenotypically normal and without clonality had none of the aforesaid atypical clinical features. CONCLUSION: Lymphoid atypia, erythrophagocytosis, loss of certain pan T-cell markers, a reduced CD4/8 ratio and TCR rearrangement define subcuticular T-cell lymphoid dyscrasia, including a subset of LEP and ILLP. The subcuticular lymphoid infiltrates represent a spectrum of histologic, immunophenotypic, and molecular abnormalities which range from those which are clearly benign to those which are clearly neoplastic, and also encompasses those cases which defy precise classification into the two aforesaid poles. PMID- 11401668 TI - Utility of HHV8 RNA detection for differentiating Kaposi's sarcoma from its mimics. AB - BACKGROUND: The diagnostic distinction of atypical vascular lesion or angiomatoid/hemosiderotic dermatofibroma and Kaposi's sarcoma can be difficult, especially in AIDS patients. Given the strong association between human herpes virus 8 (HHV8) and Kaposi's sarcoma, this study attempts to determine if HHV8 RNA detection by reverse transcription (RT) in situ polymerase chain reaction (PCR) could help in differentiating these entities. METHODS: Twenty-three Kaposi's sarcoma cases, 13 dermatofibromas, including 7 angiomatoid or hemosiderotic variants and eight atypical vascular lesions were tested for HHV8 by RT in situ PCR; five of the patients in the latter two groups were known to have AIDS. RESULTS: HHV8 RNA was detected in each of the 23 cases considered on histologic grounds to represent Kaposi's sarcoma; viral RNA and DNA localized to the majority of the endothelial and stromal spindle cells. None of the cases of dermatofibroma were HHV8 positive, whereas two of the atypical vascular lesions were viral positive, leading to a final diagnosis of Kaposi's sarcoma. CONCLUSIONS: RT in situ PCR for HHV8 RNA is a useful tool in the differential diagnosis between Kaposi's sarcoma and its mimics. PMID- 11401669 TI - Cutaneous lymphocyte-associated antigen (CLA) expression in a lymphoblastoid mantle cell lymphoma presenting with skin lesions. Comparison with other clinicopathologic presentations of mantle cell lymphoma. AB - BACKGROUND: Cutaneous lymphocyte-associated antigen (CLA) is a lymphocyte homing receptor selectively expressed by T cells of the cutaneous immune system and their malignant counterpart, that is to say, cutaneous T-cell lymphomas. However it is absent in the vast mayority of other T-cell malignancies and B-cell lineage lymphomas irrespective on primary tumor site. METHODS: Expression of CLA was investigated on six cases of mantle cell lymphoma (MCL) which differed in their histopathological subtype (typical or blastic) and their tendency to infiltrate skin and/or central nervous system (CNS). RESULTS: CLA immunostaining on neoplastic cells was only observed in a 61-year-old female suffering from a lymphoblastoid MCL which clinically presented with specific skin lesions and further developped CNS disease. In this patient, coexpression of CLA with MCL markers (CD20 and CD5) was confirmed by conventional immunohistochemistry and double immunofluorescence studies. CONCLUSIONS: To our knowledge, CLA immunoreactivity on B-cell lymphomas has not beeen previously reported. The expression of this skin-related adhesion molecule on malignant MCL cells could explain the clinical behavior of our case which presented and relapsed with cutaneous lesions. However, CLA seems not to be a MCL marker nor a CNS-related adhesion molecule. The authors review the clinical and histopathological characteristics of MCL-specific skin lesions and their diagnostic clues based on cell morphology, immunohistochemistry and molecular investigations. PMID- 11401670 TI - Cardiac epithelioid angiosarcoma presenting as cutaneous metastases. AB - BACKGROUND: Cardiac angiosarcoma is a rare tumor that has a predilection for middle-aged males and a marked predominance in the right atrium. The tumor may present abruptly with a fulminant clinical course. Initial presentation with metastatic disease is rare. Only one case has been reported of a patient who presented with cutaneous metastases. METHODS AND RESULTS: We here report the case of a 51-year-old man who initially presented with cutaneous metastases in the absence of cardiac symptoms. The skin biopsy was diagnosed as metastatic undifferentiated sarcoma. The patient died 26 days later with widely disseminated disease. At autopsy a tumor arising in the wall of the left atrium and in the interatrial septum was found. After an immunohistochemical study including CD31 and CD34 stains the diagnosis of cardiac pure epithelioid angiosarcoma was made. To the best of our knowledge this tumor variant has not been documented in the heart until now. CONCLUSION: Although cardiac angiosarcoma is a rare neoplasm, its presence should be suspected in patients with cutaneous metastatic angiosarcoma without an evident source of the tumor, even in absence of cardiac symptoms. PMID- 11401671 TI - Olmsted syndrome. AB - Olmsted syndrome is an uncommon inherited disorder of keratinization that presents mutilating palmoplantar keratoderma, perioral hyperkeratosis, leukokeratosis and alopecia. We report a case of this rare syndrome diagnosed in a 48-year-old woman and confirms the existence of a generalized abnormality in keratin expression. Immunoreactivity in our case suggests an abnormal expression of keratins 5 and 14 similar to the observed in other hyperproliferative disorders. PMID- 11401672 TI - Upregulation of P-cadherin expression in the lesional skin of pemphigus, Hailey Hailey disease and Darier's disease. AB - BACKGROUND: Autoimmune blistering diseases, pemphigus vulgaris (PV) and pemphigus foliaceus (PF), are known to be caused by binding of autoantibodies to the desmosomal cadherins, desmoglein 3 and desmoglein 1, respectively. Recently, mutations in the genes coding Ca2+ pumps leads to inherited blistering diseases, Hailey-Hailey disease (HHD) and Darier's disease (DD). Cadherins are a family of Ca2+-dependent cell adhesion molecules and P-cadherin is one of the major cadherins expressed in the epidermis. Although detailed mechanisms of acantholysis of these blistering diseases have not been fully clarified, abnormal expression of cadherins caused by altered Ca2+ concentration due to the binding of autoantibodies to cell surface or by mutations in Ca2+ pumps is suggested to be involved in mechanisms of acantholysis of these autoimmune and inherited blistering diseases. The purpose of the present study was to determine whether altered P-cadherin expression is present in these diseases. METHOD: Distribution patterns of P-cadherin in skin specimens from patients with PV (n=2), PF (n=2), HHD (n=4) and DD (n=3), were examined with confocal laser scanning microscopy using two anti-P-cadherin antibodies, 6A9 and NCC-CAD-299. RESULTS: In normal control skin, P-cadherin expression was restricted to the basal layer. In contrast, positive immunostaining of P-cadherin was observed not only in the basal cells, but also in the suprabasal cells in lesional skin of all the acantholytic diseases. CONCLUSIONS: The present results clearly demonstrated that upregulation of P-cadherin expression occurs in the acantholysis in all the four blistering diseases PV, PF, HHD and DD. Upregulation of P-cadherin may be involved in the pathomechanism of both the autoimmune blistering diseases and the inherited blistering diseases. PMID- 11401673 TI - Expression of PGP 9.5 in granular cell nerve sheath tumors: an immunohistochemical study of six cases. AB - BACKGROUND: Protein gene product 9.5 (PGP 9.5) is expressed in brain at 20 to 50 times the levels detected in other organs. Immunohistochemical studies reveal this protein is localized to both central and peripheral neurons. Recently, PGP 9.5 is reported to be a useful marker for cellular neurothekeomas. Herein we test whether PGP 9.5 is a new marker for granular cell nerve sheath tumors. MATERIAL AND METHODS: An immunohistochemical analysis for PGP 9.5 expression was carried out on all cases with the diagnosis of granular cell nerve sheath tumor seen over a 2-year period. In addition, we compared expression of PGP 9.5 with other accepted markers for neuroectodermal tumors including anti-S-100 protein and NKI/C3 monoclonal antibodies. RESULTS: Six granular cell nerve sheath tumors were diagnosed in over 80,000 dermatopathology specimens in the two-year period. These cases were all positive for PGP 9.5 as well as for S-100 protein and NK1/C3. CONCLUSION: These findings identify PGP 9.5 as a new immunohistochemical marker for use in the diagnosis of granular cell tumors. They also strengthen the histogenetic relationship between granular cell nerve sheath tumors and tumors of Schwann cell or perineurial origin. PMID- 11401674 TI - Expression of the human erythrocyte glucose transporter Glut-1 in areas of sclerotic collagen in necrobiosis lipoidica. AB - BACKGROUND: Glut-1 is the human erythrocyte glucose transporter which mediates facilitative transport of glucose across epithelial and endothelial barrier tissues. A primary abnormality in glucose transport and Glut-1 cell-surface content has been observed in fibroblasts from NIDDM and obese individuals. With the strong correlation between necrobiosis lipoidica and diabetes mellitus, we investigated the expression of Glut-1 in diabetic individuals with necrobiosis lipoidica (NL). METHODS: A polyclonal anti-Glut-1 antibody was used with a standard immunoperoxidase technique to determine Glut-1 expression by fibroblasts in areas of sclerotic collagen from specimens taken from diabetic individuals with necrobiosis lipoidica,and non-diabetic individuals with scars and granuloma annulare (GA). RESULTS: Our results showed Glut-1 expression in the areas of sclerotic collagen in patients with NL, possibly contributing to insulin resistance in these tissues. CONCLUSIONS: Our findings raises the question as to whether abnormalities in glucose transport by fibroblasts in individuals with necrobiosis lipoidica contribute to the histopathologic changes. PMID- 11401675 TI - The role of the urokinase-type plasminogen activator (uPA) and its receptor (CD87) in lipodermatosclerosis. AB - BACKGROUND: Lipodermatosclerosis refers to a sclerosing panniculitis and dermopathy of the lower extremities sometimes seen in association with venous ulceration. Matrix metalloproteinases are implicated in the pathogenesis of venous leg ulcers and the in vitro activation of recombinant MMP-2 is controlled by the plasminogen activation system. To better understand the role of plasminogen activation in the pathogenesis of venous leg ulcers we investigated fibrinolytic factors and their inhibitors in tissue samples of lipodermatolsclerosis. METHODS: The expression and the functional state of the urokinase-type plasminogen activator (uPA), the tissue-type plasminogen activator (tPA), the urokinase receptor (CD87), the plasminogen activator inhibitors-1 and 2 (PAI-1 and PAI-2) were assayed using reverse transcription polymerase chain reaction, Western blot, fibrin zymography and immunohistochemistry analyses in tissue samples of lipodermatosclerosis. RESULTS: Our results provide direct evidence of elevated expression of uPA (p<0.01) and CD87 (p<0.01) mRNA and protein level in lipodermatosclerosis in comparison with healthy skin. By immunohistochemistry, elevated expression of uPA and CD87 could be detected. Fibrin zymography showed significantly elevated endogenous uPA activity (p<0.01) in liposclerotic lesions compared to healthy controls. CONCLUSION: Our findings indicate that elevated plasminogen activation in lipodermatosclerotic tissue may play a crucial role in the pathogenesis of venous leg ulceration. PMID- 11401676 TI - Immunohistochemical expression of cyclooxygenase-2 in skin cancers. AB - BACKGROUND: Cyclooxygenase (COX), also known as prostaglandin endoperoxide synthase, catalyses the conversion of arachidonic acid to prostanoids. There are two different isoforms of COX, referred to as COX-1 and COX-2. Overexpression of COX-2 has been demonstrated in various neoplasms, such as experimentally promoted tumors, gastrointestinal cancers and breast tumors. METHODS: In this study, we used immunohistochemistry to investigate COX-2 expression in a series of basal cell epitheliomas (BCE), Bowen's disease, squamous cell carcinomas (SCC) and metastatic tumors of the skin. RESULTS: Four of 16 BCE showed a positive reaction for COX-2 and the adenoid type of BCE was the most strongly positive. In Bowen's disease, the extent of positive staining for COX-2 was even higher than that in BCE. Eleven of 15 SCC showed a positive reaction for COX-2 and the pattern of staining was heterogeneous with more intense staining in the center of the tumor nests. In metastatic tumors, the percentage of COX-2-positive tumor cells and the intensity of their staining was low compared with Bowen's disease and SCC. CONCLUSIONS: These results indicate that the intensity of COX-2 staining and its heterogeneous distribution are related to the degree of cellular differentiation and the various phenotypes of tumor cells, but the extent of COX-2 staining did not correlate with the degree of malignancy. PMID- 11401677 TI - Volar melanotic macules in a Japanese man with histopathological postinflammatory pigmentation: the volar counterpart of mucosal melanotic macules. AB - BACKGROUND: Volar melanotic macules are asymptomatic light-brown or tannish-grey macules usually found on the palms and/or soles of blacks, although they have also been reported on the volar surfaces of whites. Similar lesions have not been reported before in Japanese people. Since the cause is as yet unknown, it remains to be discussed whether they are a distinct entity. METHODS: In this report, a 52 year-old Japanese man with volar melanotic macules is reported with the clinical and histopathological findings. RESULTS: A 52-year-old Japanese man presented with many light-brown macules on his bilateral soles. He had a 20-year history of tinea pedis. Histopathological examination revealed melanophages and inflammatory infiltrates in the superficial dermis. There was a slight increase in melanin granules around the acrosyringium. Fontana-Masson stain revealed a slight increase in melanin granules in the basal layer including the acrosyringium and superficial dermis. These changes corresponded with postinflammatory pigmentation. CONCLUSIONS: This is the first report of volar melanotic macules in Japanese people. We suggest that volar melanotic macules is not an independent entity but a clinicopathological one that includes postinflammatory pigmentation, and that the condition is the volar counterpart of mucosal melanotic macules. PMID- 11401678 TI - Disseminated acanthamoebiasis presenting as lobular panniculitis with necrotizing vasculitis in a patient with AIDS. AB - BACKGROUND: Disseminated acanthamoebiasis is a rare entity, almost exclusively occurring in the immunocompromised host. METHODS: We report an unusual case of a 35-year-old female with recurrent sinusitis and multiple skin nodules demonstrating a necrotizing panniculitis, shown to be due to disseminated acanthamoebiasis. RESULTS: Histologic sections showed a neutrophilic lobular panniculitis with 20- to 30-microm trophozoites consistent with Acanthamoeba species. CONCLUSIONS: A review the literature shows that the histopathological presentation of acanthamoebiasis often eludes initial diagnostic attempts and that central nervous system (CNS) involvement is frequent and ultimately fatal. When amoebiasis is suspected, knowledge of the trophozoite and cyst forms may be helpful in distinguishing Acanthamoeba species from Entamoeba histolytica. PMID- 11401679 TI - Tumor of the follicular infundibulum with sebaceous differentiation. AB - BACKGROUND: Tumor of the follicular infundibulum (TFI) is a relatively rare tumor which clinically presents as a solitary keratotic papule usually on the head and neck which on microscopic examination typically reveals a plate-like fenestrated epithelial tumor composed of pale staining cells. METHODS: We describe a new variant of TFI. An 80-year-old male with a history of multiple basal cell carcinomas and a squamous cell carcinoma presented with a 2-year history of a red, scaly, slightly elevated plaque on the lateral aspect of his right buttock. RESULTS: Histopathological examination revealed plate-like reticulate epithelial outgrowths of large and pale cells with foci of sebaceous differentiation and numerous colloid bodies. Differential diagnosis included superficial basal cell carcinoma with sebaceous and ductal differentiation, tumor of the follicular infundibulum, an unusual fibroepithelioma of Pinkus or an eccrine fibroadenoma with sebaceous differentiation. CONCLUSION: This case illustrates a hybrid adnexal tumor with histologic features common to both tumor of the follicular infundibulum and superficial epithelioma with sebaceous differentiation. PMID- 11401680 TI - Follicular mycosis fungoides: a case report and review of the literature. AB - BACKGROUND: Follicular mycosis fungoides is an unusual variant of mycosis fungoides (MF). Unlike classic MF where atypical lymphocytes show a predilection for the epidermis (epidermotropism), follicular MF displays a malignant lymphocytic infiltrate tropic for hair follicles (folliculotropism). This malignant lymphocytic infiltrate results in follicular disruption typically manifesting clinically as plaques, comedones and follicular papules. METHODS: This report describes a 40-year-old patient with follicular MF presenting as nodules on the face and chest. Histologic examination of the patient's biopsy revealed a folliculocentric infiltrate of atypical lymphocytes with sparing of the epidermis. RESULTS AND CONCLUSIONS: Our case is discussed in the context of previously reported cases of follicular MF; we also include a review of all cases of follicular MF published to date that meet the strict criterion for this diagnosis. PMID- 11401681 TI - Pseudoepitheliomatous hyperplasia of follicular origin and malignant melanoma. PMID- 11401683 TI - Cutaneous leukocytoclastic vasculitis: the dynamic nature of the infiltrate and the expression of adhesion molecules. PMID- 11401685 TI - Co-ordinate regulation of the Escherichia coli cell cycle or The cloud of unknowing. AB - A discussion of some aspects of the regulation of chromosome replication, segregation and cell division in Escherichia coli. PMID- 11401686 TI - An analysis of the factory model for chromosome replication and segregation in bacteria. AB - Recent advances in microscopy have given us important clues as to the nature of chromosome segregation in bacteria. Most current observations favour the view that the process is co-replicational: DNA replication forks are anchored at the cell centre, and the newly replicated DNA is moved towards the cell poles. This scheme can account for orderly segregation even at high growth rates where multiple replication cycles overlap. We argue that there are five distinct activities directly involved in co-replicational segregation dynamics. These we refer to as Push, Direct, Condense, Hold and Clear. We attempt to assign one of these roles to each protein implicated in chromosome segregation. The proposed process is very different from mitosis in eukaryotic cells and perhaps more closely resembles the formation of separate sister chromatids during DNA replication. PMID- 11401687 TI - Bacterial cell division: regulating Z-ring formation. AB - The earliest stage of cell division in bacteria is the formation of a Z ring, composed of a polymer of the FtsZ protein, at the division site. Z rings appear to be synthesized in a bi-directional manner from a nucleation site (NS) located on the inside of the cytoplasmic membrane. It is the utilization of a NS specifically at the site of septum formation that determines where and when division will occur. However, a Z ring can be made to form at positions other than at the division site. How does a cell regulate utilization of a NS at the correct location and at the right time? In rod-shaped bacteria such as Escherichia coli and Bacillus subtilis, two factors involved in this regulation are the Min system and nucleoid occlusion. It is suggested that in B. subtilis, the main role of the Min proteins is to inhibit division at the nucleoid-free cell poles. In E. coli it is currently not clear whether the Min system can direct a Z ring to the division site at mid-cell or whether its main role is to ensure that division inhibition occurs away from mid-cell, a role analogous to that in B. subtilis. While the nucleoid negatively influences Z-ring formation in its vicinity in these rod-shaped organisms, the exact relationship between nucleoid occlusion and the ability to form a mid-cell Z ring is unresolved. Recent evidence suggests that in B. subtilis and Caulobacter crescentus, utilization of the NS at the division site is intimately linked to the progress of a round of chromosome replication and this may form the basis of achieving co ordination between chromosome replication and cell division. PMID- 11401688 TI - A connection between stress and development in the multicellular prokaryote Streptomyces coelicolor A3(2). AB - Morphological changes leading to aerial mycelium formation and sporulation in the mycelial bacterium Streptomyces coelicolor rely on establishing distinct patterns of gene expression in separate regions of the colony. sigmaH was identified previously as one of three paralogous sigma factors associated with stress responses in S. coelicolor. Here, we show that sigH and the upstream gene prsH (encoding a putative antisigma factor of sigmaH) form an operon transcribed from two developmentally regulated promoters, sigHp1 and sigHp2. While sigHp1 activity is confined to the early phase of growth, transcription of sigHp2 is dramatically induced at the time of aerial hyphae formation. Localization of sigHp2 activity using a transcriptional fusion to the green fluorescent protein reporter gene (sigHp2-egfp) showed that sigHp2 transcription is spatially restricted to sporulating aerial hyphae in wild-type S. coelicolor. However, analysis of mutants unable to form aerial hyphae (bld mutants) showed that sigHp2 transcription and sigmaH protein levels are dramatically upregulated in a bldD mutant, and that the sigHp2-egfp fusion was expressed ectopically in the substrate mycelium in the bldD background. Finally, a protein possessing sigHp2 promoter-binding activity was purified to homogeneity from crude mycelial extracts of S. coelicolor and shown to be BldD. The BldD binding site in the sigHp2 promoter was defined by DNase I footprinting. These data show that expression of sigmaH is subject to temporal and spatial regulation during colony development, that this tissue-specific regulation is mediated directly by the developmental transcription factor BldD and suggest that stress and developmental programmes may be intimately connected in Streptomyces morphogenesis. PMID- 11401689 TI - Pathogenicity island-dependent activation of Rho GTPases Rac1 and Cdc42 in Helicobacter pylori infection. AB - Helicobacter pylori has been identified as the major aetiological agent in the development of chronic gastritis and duodenal ulcer, and it plays a role in the development of gastric carcinoma. Attachment of H. pylori to gastric epithelial cells leads to nuclear and cytoskeletal responses in host cells. Here, we show that Rho GTPases Rac1 and Cdc42 were activated during infection of gastric epithelial cells with either the wild-type H. pylori or the mutant strain cagA. In contrast, no activation of Rho GTPases was observed when H. pylori mutant strains (virB7 and PAI) were used that lack functional type IV secretion apparatus. We demonstrated that H. pylori-induced activation of Rac1 and Cdc42 led to the activation of p21-activated kinase 1 (PAK1) mediating nuclear responses, whereas the mutant strain PAI had no effect on PAK1 activity. Activation of Rac1, Cdc42 and PAK1 represented a very early event in colonization of gastric epithelial cells by H. pylori. Rac1 and Cdc42 were recruited to the sites of bacterial attachment and are therefore probably involved in the regulation of local and overall cytoskeleton rearrangement in host cells. Finally, actin rearrangement and epithelial cell motility in H. pylori infection depended on the presence of a functional type IV secretion system encoded by the cag pathogenicity island (PAI). PMID- 11401690 TI - Signalling substitutions in the periplasmic domain of chemoreceptor Trg induce or reduce helical sliding in the transmembrane domain. AB - We used in vivo oxidative cross-linking of engineered cysteine pairs to assess conformational changes in the four-helix transmembrane domain of chemoreceptor Trg. Extending previous work, we searched for and found a fourth cross-linking pair that spanned the intrasubunit interface between transmembrane helix 1 (TM1) and its partner TM2. We determined the effects of ligand occupancy on cross linking rate constants for all four TM1-TM2 diagnostic pairs in conditions that allowed the formation of receptor-kinase complexes for the entire cellular complement of Trg. Occupancy altered all four rates in a pattern that implicated sliding of TM2 relative to TM1 towards the cytoplasm as the transmembrane signalling movement in receptor-kinase complexes. Transmembrane signalling can be reduced or induced by single amino acid substitutions in the ligand-binding region of the periplasmic domain of Trg. We determined the effects of these substitutions on conformation in the transmembrane domain and on ligand-induced changes using the diagnostic TM1-TM2 cysteine pairs. Effects on rates of in vivo cross-linking showed that induced signalling substitutions altered the relative positions of TM1 and TM2 in the same way as ligand binding, and reduced signalling substitutions blocked or attenuated the ligand-induced shift. These results provide strong support for the helical sliding model of transmembrane signalling. PMID- 11401691 TI - Different localization of SeqA-bound nascent DNA clusters and MukF-MukE-MukB complex in Escherichia coli cells. AB - MukF, MukE and MukB proteins form a complex that may participate in the organization of folded sister chromosomes in Escherichia coli. We have found that a MukB-GFPuv4 fusion protein is observed as discrete fluorescent foci, which are localized within cellular spaces occupied by nucleoids, but not at the constriction site of cell division in living cells. In contrast, MukB-GFPuv4 is distributed throughout the whole cell when either MukF or MukE is absent. Statistical analysis revealed that most newborn cells have two foci of mukB gfpUV4 at one-quarter and three-quarter positions in the cell length and one focus of SeqA-bound nascent DNA at or near the middle of the cell. Subsequently, the single SeqA focus divides into two foci, and then these migrate to the one quarter and three-quarter positions. Before cell division, most long cells have two SeqA foci and four MukB-GFPuv4 foci. In early stationary phase, SeqA foci disappear, but one or two foci of MukB-GFPuv4 remain. We discuss the reorganization and proper arrangement of folded sister chromosome in the cell quarter positions, which are performed after release from the long-time cohesion of sister chromosomes. PMID- 11401692 TI - The adhesive property of the type IV pilus-associated component PilC1 of pathogenic Neisseria is supported by the conformational structure of the N terminal part of the molecule. AB - Neisserial PilC proteins are key elements in type IV pili biogenesis and adhesion. Two pilC alleles are usually present in Neisseria meningitidis. At least one of the PilC proteins is required for pilus assembly and competence for transformation. In addition, meningococcal PilC1, but not PilC2, modulates adhesiveness, whereas, in N. gonorrhoeae, both alleles are adhesive. The meningococcal pilC genes are differently regulated, and it was shown that the expression of pilC1, but not that of pilC2, is transiently induced by bacteria cell contact. The aim of this work was to determine whether, besides regulation, PilC1-mediated adhesion was conferred by some specific protein pattern not present in the meningococcal PilC2 protein. We demonstrate first that differences within the primary sequence of the meningococcal PilC1 and PilC2 are responsible for different adhesion phenotypes, thus eliminating the regulation of transcription being solely responsible for the adhesive phenotype of PilC1. To identify the regions of PilC1 responsible for adhesion, we engineered meningococcal strains expressing various PilC1-PilC2 hybrids at the pilC1 locus. Our data demonstrate that the specific PilC1 adhesion-promoting regions are located in the amino-terminal part of the molecule and that several domains within this region probably interact with each other to promote adhesion to human cells. PMID- 11401693 TI - In vitro and in vivo nucleotide exchange directed by chimeric RNA/DNA oligonucleotides in Saccharomyces cerevisae. AB - Targeted gene repair directed by chimeric RNA/DNA oligonucleotides has proven successful in eukaryotic cells including animal and plant models. In many cases, however, there has been a disparity in the levels of gene correction or frequency. While the delivery of these chimera into the nucleus and the long-term stability or purity of these molecules may contribute to this variability, understanding the molecular regulation of conversion is the key to improving or stabilizing frequency. To this end, we have identified genes that control targeted repair, using the genetically tractable organism, Saccharomyces cerevisae and a bank of yeast mutants. Results from experiments in cell-free extracts focused our attention on RAD52, RAD1 and RAD59 as central regulatory factors. RAD1 and RAD59 appear to be required for high levels of conversion whereas RAD52 appears to act, surprisingly, in a suppressive fashion. Results from the in vitro experiments were translated into targeting experiments in vivo. Here, mutations in a fusion construct, containing a marker gene, were converted to wild type, evidenced by the expression of green fluorescence in converted cells. Because the repaired fusion gene contains a corrected neomycin sequence, cells were subsequently placed under G418 selection and conversion confirmed at the genetic level. Taken together, these results establish, for the first time, genes that participate in the regulation of targeted gene repair and provide a novel system for evaluating true frequencies of correction. Importantly, this system enables visualization of corrected (green) and uncorrected (clear) cells enabling measurements of conversion in real time. PMID- 11401694 TI - Overlapping sense and antisense transcription units in Trypanosoma brucei. AB - Procyclins are the major surface glycoproteins of insect-form Trypanosoma brucei. The procyclin expression sites are polycistronic and are transcribed by an alpha amanitin-resistant polymerase, probably RNA polymerase I (Pol I). The expression sites are flanked by transcription units that are sensitive to alpha-amanitin, which is a hallmark of Pol II-driven transcription. We have analysed a region of 9.5 kb connecting the EP/PAG2 expression site with the downstream transcription unit. The procyclin expression site is longer than was previously realized and contains an additional gene, procyclin-associated gene 4 (PAG4), and a region of unknown function, the T region, that gives rise to trans-spliced, polyadenylated RNAs containing small open reading frames (ORFs). Two new genes, GU1 and GU2, were identified in the downstream transcription unit on the opposite strand. Unexpectedly, the 3' untranslated region of GU2 and the complementary T transcripts overlap by several hundred base pairs. Replacement of GU2 by a unique tag confirmed that sense and antisense transcription occurred from a single chromosomal locus. Overlapping transcription is stage specific and may extend > or = 10 kb in insect-form trypanosomes. The nucleotide composition of the T. brucei genome is such that antisense ORFs occur frequently. If stable mRNAs can be derived from both strands, the coding potential of the genome may be substantially larger than has previously been suspected. PMID- 11401695 TI - Regulation of catalase-peroxidase (KatG) expression, isoniazid sensitivity and virulence by furA of Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis has two genes for ferric uptake regulator orthologues, one of which, furA, is situated immediately upstream of katG encoding catalase peroxidase, a major virulence factor that also activates the prodrug isoniazid. This association suggested that furA might regulate katG and other genes involved in pathogenesis. Transcript mapping showed katG to be expressed from a strong promoter, with consensus -10 and -35 elements, preceding furA. No promoter activity was demonstrated downstream of the furA start codon, using different gene reporter systems, indicating that furA and katG are co-transcribed from a common regulatory region. The respective roles of these two genes in the isoniazid susceptibility and virulence of M. tuberculosis were assessed by combinatorial complementation of a Delta(furA-katG) strain that is heavily attenuated in a mouse model of tuberculosis. In the absence of furA, katG was upregulated, cells became hypersensitive to isoniazid, and full virulence was restored, indicating that furA regulates the transcription of both genes. When furA alone was introduced into the Delta(furA-katG) mutant, survival in mouse lungs was moderately increased, suggesting that FurA could regulate genes, other than katG, that are involved in pathogenesis. These do not include the oxidative stress genes ahpC and sodA, or those for siderophore production. PMID- 11401696 TI - Conformational changes play a role in regulating the activity of the proline utilization pathway-specific regulator in Saccharomyces cerevisiae. AB - In Saccharomyces cerevisiae, the ability to use proline as a nitrogen source requires the Put3p transcriptional regulator, which turns on the expression of the proline utilization genes, PUT1 and PUT2, in the presence of the inducer proline and in the absence of preferred nitrogen sources. Changes in target gene expression occur through an alteration in activity of the DNA-bound Put3p, a member of the Zn(II)2Cys6 binuclear cluster family of proteins. Here, we report that the 'on' conformation can be mimicked in the absence of proline by the insertion of an epitope tag in several different places in the protein, as well as by specific amino acid changes that suppress a put3 mutation leading to non inducibility of the pathway. In addition, the presence of proline causes a conformational change in the Put3 protein detected by increased sensitivity to thrombin or V8 protease. These findings suggest that Put3p shifts from an inactive to an activate state via conformational changes. PMID- 11401697 TI - Heterologous expression of archaeal selenoprotein genes directed by the SECIS element located in the 3' non-translated region. AB - Previous in silico analysis of selenoprotein genes in Archaea revealed that the selenocysteine insertion (SECIS) motif necessary to recode UGA with selenocysteine was not adjacent to the UGA codon as is found in Bacteria. Rather, paralogous stem-loop structures are located in the 3' untranslated region (3' UTR), reminiscent of the situation in Eukarya. To assess the function of such putative SECIS elements, the Methanococcus jannaschii MJ0029 (fruA, which encodes the A subunit of the coenzyme F420-reducing hydrogenase) mRNA was mapped in vivo and probed enzymatically in vitro. It was shown that the SECIS element is indeed transcribed as part of the respective mRNA and that its secondary structure corresponds to that predicted by RNA folding programs. Its ability to direct selenocysteine insertion in vivo was demonstrated by the heterologous expression of MJ0029 in Methanococcus maripaludis, resulting in the synthesis of an additional selenoprotein, as analysed by 75Se labelling. The selective advantage of moving the SECIS element in the untranslated region may confer the ability to insert more than one selenocysteine into a single polypeptide. Evidence for this assumption was provided by the finding that the M. maripaludis genome contains an open reading frame with two in frame TGA codons, followed by a stem-loop structure in the 3' UTR of the mRNA that corresponds to the archaeal SECIS element. PMID- 11401698 TI - Transcription of essential cell division genes is linked to chromosome replication in Escherichia coli. AB - Cell division normally follows the completion of each round of chromosome replication in Escherichia coli. Transcription of the essential cell division genes clustered at the mra region is shown here to depend on continuing chromosomal DNA replication. After chromosome replication was blocked by either nalidixic acid treatment or thymine starvation, the transcription of these cell division genes was repressed significantly. This suggests a way in which cell division is controlled by chromosome replication. PMID- 11401699 TI - The CbrA-CbrB two-component regulatory system controls the utilization of multiple carbon and nitrogen sources in Pseudomonas aeruginosa. AB - A novel two-component system, CbrA-CbrB, was discovered in Pseudomonas aeruginosa; cbrA and cbrB mutants of strain PAO were found to be unable to use several amino acids (such as arginine, histidine and proline), polyamines and agmatine as sole carbon and nitrogen sources. These mutants were also unable to use, or used poorly, many other carbon sources, including mannitol, glucose, pyruvate and citrate. A 7 kb EcoRI fragment carrying the cbrA and cbrB genes was cloned and sequenced. The cbrA and cbrB genes encode a sensor/histidine kinase (Mr 108 379, 983 residues) and a cognate response regulator (Mr 52 254, 478 residues) respectively. The amino-terminal half (490 residues) of CbrA appears to be a sensor membrane domain, as predicted by 12 possible transmembrane helices, whereas the carboxy-terminal part shares homology with the histidine kinases of the NtrB family. The CbrB response regulator shows similarity to the NtrC family members. Complementation and primer extension experiments indicated that cbrA and cbrB are transcribed from separate promoters. In cbrA or cbrB mutants, as well as in the allelic argR9901 and argR9902 mutants, the aot-argR operon was not induced by arginine, indicating an essential role for this two-component system in the expression of the ArgR-dependent catabolic pathways, including the aruCFGDB operon specifying the major aerobic arginine catabolic pathway. The histidine catabolic enzyme histidase was not expressed in cbrAB mutants, even in the presence of histidine. In contrast, proline dehydrogenase, responsible for proline utilization (Pru), was expressed in a cbrB mutant at a level comparable with that of the wild-type strain. When succinate or other C4-dicarboxylates were added to proline medium at 1 mM, the cbrB mutant was restored to a Pru+ phenotype. Such a succinate-dependent Pru+ property was almost abolished by 20 mM ammonia. In conclusion, the CbrA-CbrB system controls the expression of several catabolic pathways and, perhaps together with the NtrB-NtrC system, appears to ensure the intracellular carbon: nitrogen balance in P. aeruginosa. PMID- 11401700 TI - Experimental genome evolution: large-scale genome rearrangements associated with resistance to replacement of a chromosomal restriction-modification gene complex. AB - Type II restriction enzymes are paired with modification enzymes that protect type II restriction sites from cleavage by methylating them. A plasmid carrying a type II restriction-modification gene complex is not easily replaced by an incompatible plasmid because loss of the former leads to cell death through chromosome cleavage. In the present work, we looked to see whether a chromosomally located restriction-modification gene complex could be replaced by a homologous stretch of DNA. We tried to replace the PaeR7I gene complex on the Escherichia coli chromosome by transducing a homologous stretch of PaeR7I modified DNA. The replacement efficiency of the restriction-modification complex was lower than expected. Some of the resulting recombinant clones retained the recipient restriction-modification gene complex as well as the homologous DNA (donor allele), and slowly lost the donor allele in the absence of selection. Analysis of their genome-wide rearrangements by Southern hybridization, inverse polymerase chain reaction (iPCR) and sequence determination demonstrated the occurrence of unequal homologous recombination between copies of the transposon IS3. It was strongly suggested that multiple rounds of unequal IS3-IS3 recombination caused large-scale duplication and inversion of the chromosome, and that only one of the duplicated copies of the recipient PaeR7I was replaced. PMID- 11401701 TI - The role of HetN in maintenance of the heterocyst pattern in Anabaena sp. PCC 7120. AB - The gene hetN encodes a putative oxidoreductase that is known to suppress heterocyst differentiation when present on a multicopy plasmid in Anabaena sp. PCC 7120. To mimic the hetN null phenotype and to examine where HetN acts in the regulatory cascade that controls heterocyst differentiation, we replaced the native chromosomal hetN promoter with the copper-inducible petE promoter. In the presence of copper, heterocyst formation was suppressed in undifferentiated filaments. When hetN expression was turned off by transferring cells to media lacking copper, the filaments initially displayed the wild-type pattern of single heterocysts but, 48 h after the induction of heterocyst formation, a pattern of multiple contiguous heterocysts predominated. Suppression of heterocyst formation by HetN appears to occur both upstream and downstream of the positive regulator HetR: overexpression of hetN in undifferentiated filaments prevents the wild-type pattern of hetR expression as well as the multiheterocyst phenotype normally observed when hetR is expressed from an inducible promoter. Green fluorescent protein fusions show that the expression of hetN in wild-type filaments normally occurs primarily in heterocysts. We propose that HetN is normally involved in the maintenance of heterocyst spacing after the initial heterocyst pattern has been established, but ectopic expression of hetN can also block the initial establishment of the pattern. PMID- 11401702 TI - Erythromycin inhibition of 50S ribosomal subunit formation in Escherichia coli cells. AB - The effects of erythromycin on the formation of ribosomal subunits were examined in wild-type Escherichia coli cells and in an RNase E mutant strain. Pulse-chase labelling kinetics revealed a reduced rate of 50S subunit formation in both strains compared with 30S synthesis, which was unaffected by the antibiotic. Growth of cells in the presence of [14C]-erythromycin showed drug binding to 50S particles and to a 50S subunit precursor sedimenting at about 30S in sucrose gradients. Antibiotic binding to the precursor correlated with the decline in 50S formation in both strains. Erythromycin binding to the precursor showed the same 1:1 stoichiometry as binding to the 50S particle. Gel electrophoresis of rRNA from antibiotic-treated organisms revealed the presence of both 23S and 5S rRNAs in the 30S region of sucrose gradients. Hybridization with a 23S rRNA-specific probe confirmed the presence of this species of rRNA in the precursor. Eighteen 50S ribosomal proteins were associated with the precursor particle. A model is presented to account for erythromycin inhibition of 50S formation. PMID- 11401704 TI - Repression of virulence genes by phosphorylation-dependent oligomerization of CsrR at target promoters in S. pyogenes. AB - csrRS encodes a two-component regulatory system that represses the transcription of a number of virulence factors in Streptococcus pyogenes, including the hyaluronic acid capsule and pyrogenic exotoxin B. CsrRS-regulated virulence factors have diverse functions during pathogenesis and are differentially expressed throughout growth. This suggests that multiple signals induce CsrRS mediated gene regulation, or that regulated genes respond differently to CsrR, or both. As a first step in dissecting the csrRS signal transduction pathway, we determined the mechanism by which CsrR mediates the repression of its target promoters. We found that phosphorylated CsrR binds directly to all but one of the promoters of its regulated genes, with different affinities. Phosphorylation of CsrR enhances both oligomerization and DNA binding. We defined the binding site of CsrR at each of the regulated promoters using DNase I and hydroxyl radical footprinting. Based on these results, we propose a model for differential regulation by CsrRS. PMID- 11401703 TI - Oligopeptide permease is required for expression of the Bacillus thuringiensis plcR regulon and for virulence. AB - PlcR is a pleiotropic regulator of virulence factors in the insect pathogen Bacillus thuringiensis and in the opportunistic human pathogen Bacillus cereus. It activates the transcription of at least 15 genes encoding extracellular proteins, including phospholipases C, proteases and enterotoxins. Expression of the plcR gene is autoregulated and activated at the onset of stationary phase. Here, we used mini-Tn10 transposition to generate a library of B. thuringiensis mutants, with the goal of characterizing genes involved in the expression of the plcR gene. Three mutant strains were identified carrying distinct mini-Tn10 insertions. The mutations impaired plcR expression and caused a deficient haemolytic phenotype, similar to the phenotype of a B. thuringiensis strain in which the plcR gene had been disrupted. The insertion sites of the three mini Tn10 transposons mapped in a five-gene operon encoding polypeptides homologous to the components of the oligopeptide permease (Opp) system of Bacillus subtilis, and with a similar structural organization. By analogy, the five B. thuringiensis genes were designated oppA, B, C, D and F. In vitro disruption of the B. thuringiensis oppB gene reproduced the effect of the mini-Tn10 insertions (i.e. the loss of haemolytic activity) and reduced the virulence of the strain against insects. These phenotypes are similar to those of a DeltaplcR mutant. Opp is required for the import of small peptides into the cell. Therefore, plcR expression might be activated at the onset of stationary phase by the uptake of a signalling peptide acting as a quorum-sensing effector. The opp mutations impaired the sporulation efficiency of B. thuringiensis when the cells were cultured in LB medium. Thus, Opp is on the pathway that ultimately regulates Spo0A phosphorylation, as is the case in B. subtilis. However, analysis of plcR expression in DeltaoppB, Deltaspo0A and DeltaoppB Deltaspo0A mutants indicates that Opp is required for plcR expression via a Spo0A-independent mechanism. PMID- 11401705 TI - Mutational analysis of the high-affinity BvgA binding site in the fha promoter of Bordetella pertussis. AB - In order to define a consensus binding sequence for the response regulator BvgA, we have undertaken a systematic analysis of contributions made by each nucleotide within the heptad half-sites that are present in an inverted orientation at the promoter for the fha operon. Using in vitro binding assays, we examined the full complement of 21 single point mutations symmetrically arranged in this heptad repeat. Both gel shift and nitrocellulose filter-binding assays provided evidence that nucleotides at positions 3 (thymidine), 4 (cytosine) and 7 (adenine) in the binding heptad contribute substantially to sequence-specific recognition by BvgA. Furthermore, a T to A conversion at position 6 reduced binding. Selected binding site mutations were introduced into a modified fha promoter and examined for their effects on BvgA activation of promoter activity in vivo. Only those substitutions most severely affecting binding in vitro affected promoter activity in vivo. The in vivo effects of substitutions that had a significant effect on binding in vitro but did not severely affect in vivo promoter activity under standard culture conditions could be detected in vivo either in combination with additional substitutions or from their effect on the sensitivity of the mutant promoters to modulation by magnesium sulphate. PMID- 11401706 TI - Heterogeneity of stress gene expression and stress resistance among individual cells of Saccharomyces cerevisiae. AB - Knowledge of gene expression and cellular responses in microorganisms is derived from analyses of populations consisting of millions of cells. Analytical techniques that provide data as population averages fail to inform of culture heterogeneity. Flow cytometry and fluorescence techniques were used to provide information on the heterogeneity of stress-responsive gene expression and stress tolerance in individual cells within populations. A sequence of DNA encoding the heat shock and stress response elements of the Saccharomyces cerevisiae HSP104 gene was used to express enhanced green fluorescent protein (EGFP). When integrated into the genome of yeast strain W303-1A, intrinsic expression of EGFP increased about twofold as cells progressed from growth on glucose to ethanol utilization in aerobic batch cultures. Staining of cells with orange/red fluorescent propidium iodide (PI), which only enters cells that have compromised membrane integrity, revealed that the population became more tolerant to 52 degrees C heat stress as it progressed from growth on glucose and through the ethanol utilization phase of aerobic batch culture. Exposure of cultures growing on glucose to a mild heat shock (shift from 25 degrees C to 37 degrees C) resulted in significantly increased expression of EGFP in the population. However, there was heterogeneity in the intensity of fluorescence of individual cells from heat-shocked cultures, indicating variability in the strength of stress response in the clonal population. Detailed analysis of the heterogeneity showed a clear positive trend between intensity of stress response and individual cell resistance, measured in terms of PI exclusion, to heat stress at 52 degrees C. Further experiments indicated that, although the mean gene expression by a population is influenced by the genetic background, the heterogeneity among individual cells in clonal populations is largely physiologically based. PMID- 11401707 TI - RNA polymerase II and TBP occupy the repressed CYC1 promoter. AB - Saccharomyces cerevisiae CYC1 gene expression has been studied in great detail with regard to the response to oxygen availability and carbon source. In the absence of oxygen and the presence of glucose, the CYC1 gene is completely repressed. Chromatin structure is thought to play an important role in CYC1 gene regulation, as nucleosome depletion results in 94-fold derepression. In addition, the CYC1 core promoter has been used extensively in hybrid constructs to study activation by heterologous transcription factors. Therefore, we set out to map the chromatin structure of the CYC1 promoter and determine its role in CYC1 gene regulation. We report here that the repressed CYC1 promoter contains no positioned nucleosomes over the core promoter. However, we did find TFIID and RNA polymerase II bound in a complex on the repressed promoter. These results indicate that recruitment of TFIID and RNA polymerase II are not rate-limiting steps in CYC1 activation. PMID- 11401708 TI - Multiple effects of protein phosphatase 2A on nutrient-induced signalling in the yeast Saccharomyces cerevisiae. AB - The trehalose-degrading enzyme trehalase is activated upon addition of glucose to derepressed cells or in response to nitrogen source addition to nitrogen-starved glucose-repressed yeast (Saccharomyces cerevisiae) cells. Trehalase activation is mediated by phosphorylation. Inactivation involves dephosphorylation, as trehalase protein levels do not change upon multiple activation/inactivation cycles. Purified trehalase can be inactivated by incubation with protein phosphatase 2A (PP2A) in vitro. To test whether PP2A was involved in trehalase inactivation in vivo, we overexpressed the yeast PP2A isoform Pph22. Unexpectedly, the moderate (approximately threefold) overexpression of Pph22 that we obtained increased basal trehalase activity and rendered this activity unresponsive to the addition of glucose or a nitrogen source. Concomitant with higher basal trehalase activity, cells overexpressing Pph22 did not store trehalose efficiently and were heat sensitive. After the addition of glucose or of a nitrogen source to starved cells, Pph22-overexpressing cells showed a delayed exit from stationary phase, a delayed induction of ribosomal gene expression and constitutive repression of stress-regulated element-controlled genes. Deletion of the SCH9 gene encoding a protein kinase involved in nutrient induced signal transduction restored glucose-induced trehalase activation in Pph22-overexpressing cells. Taken together, our results indicate that yeast PP2A overexpression leads to the activation of nutrient-induced signal transduction pathways in the absence of nutrients. PMID- 11401709 TI - Identifying regulators of transcription in an obligate intracellular pathogen: a metal-dependent repressor in Chlamydia trachomatis. AB - A prominent feature exhibited by Chlamydia trachomatis growing in an iron limiting environment is a differential pattern of protein expression. In many bacteria, iron-responsive proteins are regulated at the level of transcription by a family of repressors resembling the Escherichia coli ferric uptake regulator (Fur) protein. Although the chlamydial genome sequencing project did not unveil an obvious Fur homologue, a detailed examination indicated five unassigned open reading frames (ORFs) that would encode products with limited sequence homology to Fur. In this report, each chlamydial ORF was engineered in E. coli, and recombinant proteins were examined for functional characteristics resembling Fur. A Fur-specific polyclonal antiserum revealed that the protein encoded by ORF CT296 shares antigenic cross-recognition. Moreover, this protein forms dimers in solution in a fashion analogous to E. coli Fur. Further studies confirmed that the product of ORF CT296 is able to (i) complement Fur activity in a mutant strain of E. coli; and (ii) specifically bind to a 19 bp consensus sequence found in promoters of iron-regulated genes in E. coli. We propose a designation of dcrA (divalent cation-dependent regulator A) for ORF CT296, which encodes a protein distantly related to E. coli Fur. DcrA represents the first repressor described for this obligate intracellular bacterium. PMID- 11401710 TI - Variations in the surface proteins and restriction enzyme systems of Mycoplasma pulmonis in the respiratory tract of infected rats. AB - Restriction and modification (R-M) systems are generally thought to protect bacteria from invasion by foreign DNA. This paper proposes the existence of an alternative role for the phase-variable R-M systems encoded by the hsd loci of Mycoplasma pulmonis. Populations of M. pulmonis cells that arose during growth in different environments were compared with respect to R-M activity and surface antigen production. When M. pulmonis strain X1048 was propagated in laboratory culture medium, > 95% of colony-forming units (cfu) lacked R-M activity and produced the variable surface protein VsaA. Mycoplasmas isolated from the nose of experimentally infected rats also lacked R-M activity and produced VsaA. In contrast, the cell population of mycoplasmas isolated from the lower respiratory tract of the infected rats was more complex. The most dramatic results were obtained for mycoplasmas isolated from the trachea. At 14 days postinfection, 38% of mycoplasma isolates produced a Vsa protein other than VsaA, and 34% of isolates had active restriction systems. These data suggest that differences in selection pressures in animal tissues affect the surface proteins and the R-M activity of the mycoplasmal cell population. We propose that variations in the production of R-M activity and cell surface proteins are important for the survival of the mycoplasma within the host. PMID- 11401711 TI - Sortase-catalysed anchoring of surface proteins to the cell wall of Staphylococcus aureus. AB - Many surface proteins of Gram-positive bacteria are anchored to the cell wall envelope by a transpeptidation mechanism, requiring a C-terminal sorting signal with a conserved LPXTG motif. Sortase, a membrane protein of Staphylococcus aureus, cleaves polypeptides between the threonine and the glycine of the LPXTG motif and catalyses the formation of an amide bond between the carboxyl-group of threonine and the amino-group of peptidoglycan cross-bridges. S. aureus mutants lacking the srtA gene fail to anchor and display some surface proteins and are impaired in the ability to cause animal infections. Sortase acts on surface proteins that are initiated into the secretion (Sec) pathway and have their signal peptide removed by signal peptidase. The S. aureus genome encodes two sets of sortase and secretion genes. It is conceivable that S. aureus has evolved more than one pathway for the transport of 20 surface proteins to the cell wall envelope. PMID- 11401712 TI - Transcriptional control of the GAL/MEL regulon of yeast Saccharomyces cerevisiae: mechanism of galactose-mediated signal transduction. AB - In the yeast Saccharomyces cerevisiae, the interplay between Gal3p, Gal80p and Gal4p determines the transcriptional status of the genes needed for galactose utilization. The interaction between Gal80p and Gal4p has been studied in great detail; however, our understanding of the mechanism of Gal3p in transducing the signal from galactose to Gal4p has only begun to emerge recently. Historically, Gal3p was believed to be an enzyme (catalytic model) that converts galactose to an inducer or co-inducer, which was thought to interact with GAL80p, the repressor of the system. However, recent genetic analyses indicate an alternative 'protein-protein interaction model'. According to this model, Gal3p is activated by galactose, which leads to its interaction with Gal80p. Biochemical and genetic experiments that support this model provided new insights into how Gal3p interacts with the Gal80p-Gal4p complex, alleviates the repression of Gal80p and thus allows Gal4p to activate transcription. Recently, a galactose-independent signal was suggested to co-ordinate the induction of GAL genes with the energy status of the cell. PMID- 11401713 TI - The Saccharomyces cerevisiae Sko1p transcription factor mediates HOG pathway dependent osmotic regulation of a set of genes encoding enzymes implicated in protection from oxidative damage. AB - A major part of the transcriptional response of yeast cells to osmotic shock is controlled by the HOG pathway and several downstream transcription factors. Sko1p is a repressor that mediates HOG pathway-dependent regulation by binding to CRE sites in target promoters. Here, we report five target genes of Hog1p-Sko1p: GRE2, AHP1, SFA1, GLR1 and YML131w. The two CREs in the GRE2 promoter function as activating sequences and, hence, bind (an) activator protein(s). However, the two other yeast CRE-binding proteins, Aca1p and Aca2p, are not involved in regulation of the GRE2 promoter under osmotic stress. In the absence of the co-repressor complex Tup1p-Ssn6p/Cyc8p, which is recruited by Sko1p, stimulation by osmotic stress is still observed. These data indicate that Sko1p is not only required for repression, but also involved in induction upon osmotic shock. All five Sko1p targets encode oxidoreductases with demonstrated or predicted roles in repair of oxidative damage. Altered basal expression levels of these genes in hog1Delta and sko1Delta mutants may explain the oxidative stress phenotypes of these mutants. All five Sko1p target genes are induced by oxidative stress, and induction involves Yap1p. Although Sko1p and Yap1p appear to mediate osmotic and oxidative stress responses independently, Sko1p may affect Yap1p promoter access or activity. The five Sko1p target genes described here are suitable models for studying the interplay between osmotic and oxidative responses at the molecular and physiological levels. PMID- 11401714 TI - SUT1p interaction with Cyc8p(Ssn6p) relieves hypoxic genes from Cyc8p-Tup1p repression in Saccharomyces cerevisiae. AB - SUT1 is a hypoxic gene encoding a nuclear protein that belongs to the Zn[II]2Cys 6 family. It has been shown that constitutive expression of SUT1 induces exogenous sterol uptake in aerobically growing Saccharomyces cerevisiae cells. A differential display approach was used to identify genes whose transcription is modified upon SUT1 induction. Within the promoter sequence of one of these genes, DAN1, we identified the region responsive to SUT1 and showed that it has a strong repressive activity when cloned in the vicinity of distinct promoters. Upon SUT1 constitutive expression in aerobiosis, the repression is released, allowing enhanced transcription of the reporter gene. We provide evidence that the repression is promoted by the Cyc8p(Ssn6p)-Tup1p co-repressor and that release of repression is the result of a physical interaction between Sut1p and Cyc8p. Moreover, genetic data suggest that complete derepression of the reporter gene requires a functional Cyc8p. In addition, we show that Sut1p is involved in the induction of hypoxic gene transcription when the cells are shifted from aerobiosis to anaerobiosis. PMID- 11401715 TI - Protein regions important for plasminogen activation and inactivation of alpha2 antiplasmin in the surface protease Pla of Yersinia pestis. AB - The plasminogen activator, surface protease Pla, of the plague bacterium Yersinia pestis is an important virulence factor that enables the spread of Y. pestis from subcutaneous sites into circulation. Pla-expressing Y. pestis and recombinant Escherichia coli formed active plasmin in the presence of the major human plasmin inhibitor, alpha2-antiplasmin, and the bacteria were found to inactivate alpha2 antiplasmin. In contrast, only poor plasminogen activation and no cleavage of alpha2-antiplasmin was observed with recombinant bacteria expressing the homologous gene ompT from E. coli. A beta-barrel topology model for Pla and OmpT predicted 10 transmembrane beta-strands and five surface-exposed loops L1-L5. Hybrid Pla-OmpT proteins were created by substituting each of the loops between Pla and OmpT. Analysis of the hybrid molecules suggested a critical role of L3 and L4 in the substrate specificity of Pla towards plasminogen and alpha2 antiplasmin. Substitution analysis at 25 surface-located residues showed the importance of the conserved residues H101, H208, D84, D86, D206 and S99 for the proteolytic activity of Pla-expressing recombinant E. coli. The mature alpha-Pla of 292 amino acids was processed into beta-Pla by an autoprocessing cleavage at residue K262, and residues important for the self-recognition of Pla were identified. Prevention of autoprocessing of Pla, however, had no detectable effect on plasminogen activation or cleavage of alpha2-antiplasmin. Cleavage of alpha2-antiplasmin and plasminogen activation were influenced by residue R211 in L4 as well as by unidentified residues in L3. OmpT, which is not associated with invasive bacterial disease, was converted into a Pla-like protease by deleting residues D214 and P215, by substituting residue K217 for R217 in L4 of OmpT and also by substituting the entire L3 with that from Pla. This simple modification of the surface loops and the substrate specificity of OmpT exemplifies the evolution of a housekeeping protein into a virulence factor by subtle mutations at critical protein regions. We propose that inactivation of alpha2-antiplasmin by Pla of Y. pestis promotes uncontrolled proteolysis and contributes to the invasive character of plague. PMID- 11401716 TI - The Legionella pneumophila IcmR protein exhibits chaperone activity for IcmQ by preventing its participation in high-molecular-weight complexes. AB - A key event in legionellosis is the ability of Legionella pneumophila to survive and proliferate inside alveolar macrophages. The dot/icm genes, which are necessary for intracellular growth, show sequence similarity to genes encoding conjugative transfer systems, and it is believed that they are responsible for the formation of a secretion apparatus. Evidence is provided here that the IcmR and IcmQ proteins participate in a chaperone-substrate relationship similar to that observed for translocated proteins in type III and type IV secretion apparatuses. Immobilized IcmQ was found to bind IcmR from crude bacterial extracts efficiently. Furthermore, purified IcmR and IcmQ bind with high affinity. This interaction was also observed in vivo by co-immunoprecipitation. The presence of IcmR was found to affect the physical state of IcmQ directly. In the absence of IcmR, IcmQ formed high-molecular-weight complexes both in vivo and in vitro, whereas IcmR prevented and reversed the formation of these complexes. PMID- 11401717 TI - Visualization of secreted Hrp and Avr proteins along the Hrp pilus during type III secretion in Erwinia amylovora and Pseudomonas syringae. AB - Pili are required for protein and/or DNA transfer from bacteria to recipient plant or bacterial cells, based on genetic evidence. However, it has never been shown directly that the effector proteins or DNA are localized along or inside the pili in situ. Failure to visualize an association of effector proteins/DNA with pili is the central issue in the debate regarding the exact function of pili in protein and DNA transfer. In this study, a newly developed in situ immunogold labelling procedure enabled visualization of the specific localization of type III effector proteins of Erwinia amylovora and Pseudomonas syringae pv. tomato along the Hrp pilus, but not along the flagellum or randomly in the intercellular space. In contrast, PelE, a pectate lyase secreted via the type II protein secretion system, was not associated with the Hrp pilus. These results provide direct evidence that type III secretion occurs only at the site of Hrp pilus assembly and that the Hrp pilus guides the transfer of effector proteins outside the bacterial cell, favouring the 'conduit/guiding filament' model. PMID- 11401718 TI - Systematic mutagenesis of the DNA binding sites for SoxS in the Escherichia coli zwf and fpr promoters: identifying nucleotides required for DNA binding and transcription activation. AB - SoxS is the direct transcriptional activator of at least 15 genes of the Escherichia coli superoxide regulon. SoxS is small (107 amino acids), binds DNA as a monomer and recognizes a highly degenerate DNA binding site, termed 'soxbox'. Like other members of the AraC/XylS family, SoxS has two putative helix turn-helix (HTH) DNA-binding motifs, and it has been proposed that each HTH motif recognizes a highly conserved recognition element of the soxbox. To determine which nucleotides are important for SoxS binding, we conducted a systematic mutagenesis of the DNA binding sites for SoxS in the zwf and fpr promoters and determined the effect of the soxbox mutations on SoxS DNA binding and transcription activation in vivo by measuring beta-galactosidase activity in strains with fusions to lacZ. We found that the sequences GCAC and CAAA, termed recognition elements 1 and 2 (RE 1 and RE 2), respectively, are critical for SoxS binding, as mutations within these elements severely hinder or eliminate SoxS dependent transcription activation; substitutions within RE 2 (CAAA), however, are tolerated better than changes within RE 1 (GCAC). Although substitutions at the seven positions separating the two REs had only a modest effect on SoxS binding, AT basepairs were favoured within this 'spacer' region, presumably because, by facilitating DNA bending, they help bring the two recognition elements into proper juxtaposition. We also found that the 'invariant A' present at position 1 of 14/15 functional soxboxes identified thus far is important for SoxS binding, as a change to any other nucleotide at this position reduced SoxS dependent transcription by approximately 50%. In addition, positions surrounding the REs seem to show a context effect, in that certain substitutions there have little or no effect when the RE has the optimal binding sequence, but produce a pronounced effect when the RE has a suboptimal sequence. We propose that these nucleotides play an important role in effecting differential expression from the various promoters. Lastly, we used gel retardation assays to show that alterations in transcription activation in vivo are caused by effects on DNA binding. Based on this exhaustive mutagenesis, we propose the following optimal sequence for SoxS binding: AnVGCACWWWnKRHCAAAHn (n = A, C, G, T; V = A, C, G; W = A, T; K = G, T; R = A, G; H = A, C, T). PMID- 11401719 TI - Poly(A) polymerase activity and RNA polyadenylation in Streptomyces coelicolor A3(2). AB - The Streptomyces coelicolor genome sequence was searched for open reading frames (ORFs) similar to Escherichia coli poly(A) polymerase I, revealing an ORF with 36% amino acid sequence identity to that protein. Mycelial extracts prepared from S. coelicolor cultures incorporated radioactive ATP into an acid-insoluble form, and some of the products of this incorporation had the properties expected of poly(A). [3H]-uridine and [3H]-adenosine were used to label the RNA in S. coelicolor cultures of different ages, and total RNA was fractionated by oligo dT cellulose chromatography. Approximately 3% of the total uridine-labelled RNA and 11% of the adenosine-labelled RNA were retained by the oligo dT cellulose columns. Enzymatic digestion of the retained RNA supported the conclusion that a significant fraction of the adenosine label was present in 3'-poly(A) chains. Measurement of poly(A) tail lengths by end labelling of total RNA and RNase digestion revealed a maximum length of approximately 18 residues. Radioactive cDNA prepared from the RNA fraction retained by oligo dT cellulose hybridized to the 16S and 23S genes from a streptomycete ribosomal RNA operon but not to the 5S gene. Reverse transcription-polymerase chain reaction (RT-PCR) revealed the presence of mRNAs in the RNA fraction retained by oligo dT cellulose. PMID- 11401720 TI - The Candida glabrata Amt1 copper-sensing transcription factor requires Swi/Snf and Gcn5 at a critical step in copper detoxification. AB - The yeast Candida glabrata rapidly autoactivates transcription of the AMT1 gene in response to potentially toxic copper levels through the copper-inducible binding of the Amt1 transcription factor to a metal response element (MRE) within a positioned nucleosome. Our previous studies have characterized the role of a 16 bp homopolymeric dA:dT DNA structural element in facilitating rapid Amt1 access to the AMT1 promoter nucleosomal MRE. In this study, we have used the genetically more facile yeast Saccharomyces cerevisiae to identify additional cellular factors that are important for promoting rapid autoactivation of the AMT1 gene in response to toxic copper levels. We demonstrate that the Swi/Snf nucleosome remodelling complex and the histone acetyltransferase Gcn5 are both essential for AMT1 gene autoregulation, and that the requirement for these chromatin remodelling factors is target gene specific. Chromatin accessibility measurements performed in vitro and in vivo indicate that part of the absolute requirement for these factors is derived from their involvement in facilitating nucleosomal access to the AMT1 promoter MRE. Additionally, these data implicate the involvement of Swi/Snf and Gcn5 at multiple levels of AMT1 gene autoregulation. PMID- 11401721 TI - Accumulation of manganese in Neisseria gonorrhoeae correlates with resistance to oxidative killing by superoxide anion and is independent of superoxide dismutase activity. AB - As a facultative aerobe with a high iron requirement and a highly active aerobic respiratory chain, Neisseria gonorrhoeae requires defence systems to respond to toxic oxygen species such as superoxide. It has been shown that supplementation of media with 100 microM Mn(II) considerably enhanced the resistance of this bacterium to oxidative killing by superoxide. This protection was not associated with the superoxide dismutase enzymes of N. gonorrhoeae. In contrast to previous studies, which suggested that some strains of N. gonorrhoeae might not contain a superoxide dismutase, we identified a sodB gene by genome analysis and confirmed its presence in all strains examined by Southern blotting, but found no evidence for sodA or sodC. A sodB mutant showed very similar susceptibility to superoxide killing to that of wild-type cells, indicating that the Fe-dependent SOD B did not have a major role in resistance to oxidative killing under the conditions tested. The absence of a sodA gene indicated that the Mn-dependent protection against oxidative killing was independent of Mn-dependent SOD A. As a sodB mutant also showed Mn-dependent resistance to oxidative killing, then it is concluded that this resistance is independent of superoxide dismutase enzymes. Resistance to oxidative killing was correlated with accumulation of Mn(II) by the bacterium. We hypothesize that this bacterium uses Mn(II) as a chemical quenching agent in a similar way to the already established process in Lactobacillus plantarum. A search for putative Mn(II) uptake systems identified an ABC cassette-type system (MntABC) with a periplasmic-binding protein (MntC). An mntC mutant was shown to have lowered accumulation of Mn(II) and was also highly susceptible to oxidative killing, even in the presence of added Mn(II). Taken together, these data show that N. gonorrhoeae possesses a Mn(II) uptake system that is critical for resistance to oxidative stress. PMID- 11401722 TI - Molecular characterization of the surface layer proteins from Clostridium difficile. AB - Many bacteria express a surface-exposed proteinaceous layer, termed the S-layer, which forms a regular two-dimensional array visible by electron microscopy. Clostridium difficile is unusual in expressing two S-layer proteins (SLPs), which are of varying size in a number of strains. In an approach combining molecular biology with mass spectrometric sequencing strategies, we have identified the structural gene (slpA) for the S-layer from three strains of C. difficile. Both proteins are derived from a common precursor, and processing involves the removal of a signal peptide and a second cleavage to release the two mature SLPs. To our knowledge, this is the first example in which two SLPs have been shown to derive from a single gene product through post-translational processing, rather than from the expression of separate genes. The higher molecular weight (MW) SLP is highly conserved among the three strains, whereas the lower MW SLP shows considerable sequence diversity, reflecting the results from Western blotting. The high-MW SLP shows weak homology to N-acetyl muramoyl-L-alanine amidase from Bacillus subtilis, and both the native SLP from C. difficile and a recombinant protein expressed in Escherichia coli were found to display amidase activity by zymography. The high-MW SLPs showed evidence of glycosylation, whereas the lower MW proteins did not. A family of genes with sequence homology to the amidase domain of the high-MW SLP was identified in the C. difficile strain 630 genome, some of which are located in the same region of the genome as slpA and were shown by reverse transcription-polymerase chain reaction (RT-PCR) analysis to be transcribed. PMID- 11401723 TI - RpoS co-operates with other factors to induce Legionella pneumophila virulence in the stationary phase. AB - Legionella pneumophila replicates within amoebae and macrophages and causes the severe pneumonia Legionnaires' disease. When broth cultures enter the post exponential growth (PE) phase or experience amino acid limitation, L. pneumophila accumulates the stringent response signal (p)ppGpp and expresses traits likely to promote transmission to a new phagocyte. The hypothesis that a stringent response mechanism regulates L. pneumophila virulence was bolstered by our finding that the avirulent mutant Lp120 contains an internal deletion in the gene encoding the stationary phase sigma factor RpoS. To test directly whether RpoS co-ordinates virulence with stationary phase, isogenic wild-type, rpoS-120 and rpoS null mutant strains were constructed and analysed. PE phase L. pneumophila became cytotoxic by an RpoS-independent pathway, but their sodium sensitivity and maximal expression of flagellin required RpoS. Likewise, full induction of sodium sensitivity by experimentally induced (p)ppGpp synthesis required RpoS. To replicate efficiently in macrophages, L. pneumophila used both RpoS-dependent and -independent pathways. Like those containing the dotA type IV secretory apparatus mutant, phagosomes harbouring either rpoS or dotA rpoS mutants rapidly acquired the late endosomal protein LAMP-1, but not the lysosomal marker Texas red ovalbumin. Together, the data support a model in which RpoS co-operates with other regulators to induce L. pneumophila virulence in the PE phase. PMID- 11401724 TI - Pseudomonas aeruginosa GacA, a factor in multihost virulence, is also essential for biofilm formation. AB - We have investigated a potential role for GacA, the response regulator of the GacA/GacS two-component regulatory system, in Pseudomonas aeruginosa biofilm formation. When gacA was disrupted in strain PA14, a 10-fold reduction in biofilm formation capacity resulted relative to wild-type PA14. However, no significant difference was observed in the planktonic growth rate of PA14 gacA(-). Providing gacA in trans on the multicopy vector pUCP-gacA abrogated the biofilm formation defect. Scanning electron microscopy of biofilms formed by PA14 gacA(-) revealed diffuse clusters of cells that failed to aggregate into microcolonies, implying a deficit in biofilm development or surface translocation. Motility assays revealed no decrease in PA14 gacA(-) twitching or swimming abilities, indicating that the defect in biofilm formation is independent of flagellar-mediated attachment and solid surface translocation by pili. Autoinducer and alginate bioassays were performed similarly, and no difference in production levels was observed, indicating that this is not merely an upstream effect on either quorum sensing or alginate production. Antibiotic susceptibility profiling demonstrated that PA14 gacA(-) biofilms have moderately decreased resistance to a range of antibiotics relative to PA14 wild type. This study establishes GacA as a new and independent regulatory element in P. aeruginosa biofilm formation. PMID- 11401725 TI - Pleiotropic transcriptional repressor CodY senses the intracellular pool of branched-chain amino acids in Lactococcus lactis. AB - Proteolysis is essential for supplying Lactococcus lactis with amino acids during growth in milk. Expression of the major components of the L. lactis proteolytic system, including the cell wall proteinase (PrtP), the oligopeptide transport system (Opp) and at least four intracellular peptidases (PepO1, PepN, PepC, PepDA2), was shown previously to be controlled negatively by a rich nitrogen source. The transcription of prtP, opp-pepO1, pepN and pepC genes is regulated by dipeptides in the medium. Random insertion mutants derepressed for nitrogen control in the expression of the oligopeptide transport system were isolated using an opp-lacZ fusion. A third of the mutants were targeted in the same locus. The product of the inactivated gene shared 48% identity with CodY from Bacillus subtilis, a pleiotropic repressor of the dipeptide permease operon (dpp) and several genes including genes involved in amino acid degradation and competence induction. The signal controlling CodY-dependent repression was searched for by analysing the response of the opp-lux fusion to the addition of 67 dipeptides with different amino acid compositions. Full correlation was found between the dipeptide content in branched-chain amino acids (BCAA; isoleucine, leucine or valine) and their ability to mediate the repression of opp-pepO1 expression. The repressive effect resulting from specific regulatory dipeptides was abolished in L. lactis mutants affected in terms of their transport or degradation into amino acids, showing that the signal was dependent on the BCAA pool in the cell. Lastly, the repression of opp-pepO1 expression was stronger in a mutant unable to degrade BCAAs, underlining the central role of BCAAs as a signal for CodY activity. This pattern of regulation suggests that, in L. lactis and possibly other Gram-positive bacteria, CodY is a pleiotropic repressor sensing nutritional supply as a function of the BCAA pool in the cell. PMID- 11401726 TI - Modulation of intracellular calcium and proliferative activity of invertebrate and vertebrate cells by ethylene. AB - BACKGROUND: Ethylene is a widely distributed alkene product which is formed enzymatically (e.g., in plants) or by photochemical reactions (e.g., in the upper oceanic layers from dissolved organic carbon). This gaseous compound was recently found to induce in cells from the marine sponge Suberites domuncula, an increase in intracellular Ca2+ level ([Ca2+]i) and an upregulation of the expression of two genes, the potential ethylene-responsive gene, SDERR, and a Ca2+/calmodulin dependent protein kinase. RESULTS: Here we describe for the first time, that besides sponge cells, mammalian cell lines (mouse NIH-3T3 and human HeLa and SaOS 2 cells) respond to ethylene, generated by ethephon, with an immediate and strong, transient increase in [Ca2+]i level, as demonstrated using Fura-2 imaging method. A rise of [Ca2+]i level was also found following exposure to ethylene gas of cells kept under pressure (SaOS-2 cells). The upregulation of [Ca2+]i was associated with an increase in the level of the cell cycle-associated Ki-67 antigen. In addition, we show that the effect of ethephon addition to S. domuncula cells depends on the presence of calcium in the extracellular milieu. CONCLUSION: The results presented in this paper indicate that ethylene, previously known to act as a mediator (hormone) in plants only, deserves also attention as a potential signaling molecule in higher vertebrates. Further studies are necessary to clarify the specificity and physiological significance of the effects induced by ethylene in mammalian cells. PMID- 11401727 TI - Dimerization of receptor protein-tyrosine phosphatase alpha in living cells. AB - BACKGROUND: Dimerization is an important regulatory mechanism of single membrane spanning receptors. For instance, activation of receptor protein-tyrosine kinases (RPTKs) involves dimerization. Structural, functional and biochemical studies suggested that the enzymatic counterparts of RPTKs, the receptor protein-tyrosine phosphatases (RPTPs), are inhibited by dimerization, but whether RPTPs actually dimerize in living cells remained to be determined. RESULTS: In order to assess RPTP dimerization, we have assayed Fluorescence Resonance Energy Transfer (FRET) between chimeric proteins of cyan- and yellow-emitting derivatives of green fluorescent protein, fused to RPTPalpha, using three different techniques: dual wavelength excitation, spectral imaging and fluorescence lifetime imaging. All three techniques suggested that FRET occurred between RPTPalpha -CFP and -YFP fusion proteins, and thus that RPTPalpha dimerized in living cells. RPTPalpha dimerization was constitutive, extensive and specific. RPTPalpha dimerization was consistent with cross-linking experiments, using a non-cell-permeable chemical cross-linker. Using a panel of deletion mutants, we found that the transmembrane domain was required and sufficient for dimerization. CONCLUSIONS: We demonstrate here that RPTPalpha dimerized constitutively in living cells, which may be mediated by the transmembrane domain, providing strong support for the model that dimerization is involved in regulation of RPTPs. PMID- 11401728 TI - Imiquimod for the treatment of genital warts: a quantitative systematic review. AB - OBJECTIVE: To review published randomised controlled trials to assess the benefit and harm of imiquimod in the treatment of external genital warts. DATA SOURCES: MEDLINE (1966 - December 2000), Cochrane Library (Issue 3, 2000) and PubMed (December 15, 2000), review articles and reference lists. REVIEW METHODS: Included studies had to be randomised trials of imiquimod, to be full published papers, and to have a comparison group. Quality of trial reporting was assessed. Relative benefit and number needed to treat were calculated for the main outcomes of wart clearance at the end of therapy, of at least 50% reduction in wart area, and of complete clearance at the end of treatment and no recurrence of warts during a follow-up period, as well as for adverse effect withdrawal or lack of efficacy withdrawal. RESULTS: There were six trials, all with quality scores of 3 (out of 5) or greater. In five trials with HIV-negative patients complete clearance of warts at the end of treatment occurred in 51% of patients treated with imiquimod 2% or 5% cream and 6% of placebo treated patients. The number needed to treat was 2.2 (95% confidence interval 2.0 to 2.6). In four trials at least 50% wart area reduction occurred with 72% of patients treated with imiquimod 5% cream and 20% of placebo treated patients. The number needed to treat was 1.9 (1.7 to 2.2). In three trials complete clearance of warts at the end of treatment plus no recurrence occurred in 37% of patients treated with imiquimod 5% cream and 4% of those treated with placebo. The number needed to treat was 3.0 (2.5 to 3.8). Adverse event withdrawal was rare and no more likely with imiquimod than with placebo. Imiquimod was not effective in one trial in HIV positive patients. CONCLUSION: The evidence base for imiquimod in treating genital warts is of high quality and the necessary size from which to draw useful conclusions. Imiquimod is effective in home application, though not in patients with HIV infection with the evidence presently available. PMID- 11401729 TI - Counting the costs: comparing depot medroxyprogesterone acetate and norethisterone oenanthate utilisation patterns in South Africa. AB - BACKGROUND: In South Africa, where health care resources are limited, it is important to ensure that drugs provision and use is rational. The Essential Drug List includes depot medroxyprogesterone acetate (DMPA) and norethisterone oenanthate (NET-EN) as injectable progestagen-only contraceptives (IPCs), and both products are extensively used. OBJECTIVES AND METHODS: Utilisation patterns of the injectable contraceptive products DMPA and NET-EN are compared in the context of current knowledge of the safety and efficacy of these agents. Utilisation patterns were analysed by means of a Pareto (ABC) analysis of IPCs issued from 4 South African provincial pharmaceutical depots over 3 financial years. A case study from rural KwaZulu-Natal, South Africa, is used to examine utilisation patterns and self-reported side effects experienced by 187 women using IPCs. RESULTS: IPCs accounted for a substantial share of total state expenditure on drugs. While more DMPA than NET-EN was issued, NET-EN distribution from 2 depots increased over the 3-year period. Since DMPA was cheaper, if all NET-EN clients in the 1999/2000 financial year (annualised) had used DMPA, the 4 depots could have saved 4.95 million South African Rands on product acquisition costs alone. The KZN case study showed slightly more NET-EN (54%) than DMPA (46%) use; no significant differences in self-reported side effects; and that younger women were more likely to use NET-EN than DMPA (p = 0.0001). CONCLUSIONS: Providing IPCs on the basis of age is not appropriate or cost effective. Rational use of these products should include consideration of the cost of prescribing one over another. PMID- 11401730 TI - Are antibiotics appropriate for the treatment of acute sinusitis in adults? PMID- 11401731 TI - How accurate is a second-trimester prenatal ultrasound in the diagnosis of Down syndrome? PMID- 11401732 TI - Should patients with nonulcer dyspepsia and Helicobacter pylori be treated with antibiotics? PMID- 11401733 TI - Does treatment with topical metronidazole improve seborrheic dermatitis? PMID- 11401734 TI - Clinical inquiries. Are antibiotics effective for travelers' diarrhea? PMID- 11401735 TI - Clinical inquiries. What is the differential diagnosis of an elevated alkaline phosphatase (AP) level in an otherwise asymptomatic patient? PMID- 11401736 TI - Beta-blockers to reduce mortality in patients with systolic dysfunction: a meta analysis. AB - OBJECTIVE: The researchers reviewed published clinical trials and performed a meta-analysis to assess if therapy with adrenergic beta-antagonists (beta blockers) reduces the risk of mortality in patients with systolic dysfunction. STUDY DESIGN: A systematic review was performed with meta-analysis where appropriate. Clinical trials were reviewed with respect to the quality of the research methods, including patient population and end points. Two independent reviewers calculated relative risk, relative risk reduction, absolute risk reduction, and number needed to treat for the total mortality end point reported in each trial. A meta-analysis was performed. DATA SOURCES: The study team searched pertinent indexing services and references from published articles for relevant literature. The selected clinical trials were randomized, double blinded, and controlled, and included patients with systolic heart failure. Mortality was assessed as a primary or secondary end point. OUTCOMES MEASURED: The primary outcome was mortality. RESULTS: Statistically and clinically significant improvement, including a statistically significant reduction in mortality, has been noted in patients receiving therapy with either bisoprolol, carvedilol, or metoprolol. Pooled analysis revealed a statistically significant reduction in the risk of total mortality (odds ratio [OR]MH=0.66; 95% confidence interval [CI], 0.58-0.75) and sudden death (ORMH=0.61; 95% CI, 0.5-0.75) for patients receiving beta-blocker therapy. CONCLUSIONS: All patients with New York Heart Association class II and III heart failure should receive beta-blocker therapy with bisoprolol, carvedilol, or metoprolol. Additional clinical trials are ongoing and will provide further data on which patients receive the greatest benefit from therapy and which beta-blocker may be preferred. PMID- 11401737 TI - Weight management: what patients want from their primary care physicians. AB - OBJECTIVE: The researchers wanted to determine the weight management experiences of patients in primary care, and what those patients want from their physicians. STUDY DESIGN: Patients completed a survey in a primary care waiting room. Afterward they were measured for body mass index (BMI). POPULATION: A total of 410 consecutive adult patients in 2 primary care practices at the University of California, San Francisco, were approached, and 366 (89%) completed the survey. OUTCOMES MEASURED: The primary outcomes were patient attitudes about weight loss, previous weight management experiences with their current physicians, and future preferences for weight management within the primary care relationship. RESULTS: Ninety-seven percent of the obese patients (BMI > 30), 84% of the overweight patients (BMI=25-30), and 39% of the non-overweight patients (BMI < 25) thought they needed to lose weight. Forty-nine percent of the obese patients, 24% of the overweight patients, and 12% of the non-overweight patients had discussed weight with their current physicians. The types of weight management assistance that patients most wanted from their physicians were: (1) dietary advice, (2) help with setting realistic weight goals, and (3) exercise recommendations. CONCLUSIONS: Although most patients believe they should lose weight, this is often not discussed during office visits. Most patients (especially those who are overweight or obese) want more help with weight management than they are getting from their primary care physicians. PMID- 11401738 TI - Obesity management in primary care: changing the status quo. PMID- 11401739 TI - The patient's perspective of irritable bowel syndrome. AB - OBJECTIVE: The researchers wanted to understand how irritable bowel syndrome (IBS) affects patients' lives and their interactions with physicians and the health care system. STUDY DESIGN: A qualitative study was performed using focus groups of people with physician-diagnosed IBS. Immersion/crystallization was used to identify overriding themes. POPULATION: Adult volunteers with a previous physician diagnosis of IBS were included. OUTCOMES MEASURED: The outcomes were patient-reported symptoms, episode triggers, treatments, lifestyle changes, and interactions with their physicians that were related to IBS, and overriding themes identified from the focus groups. RESULTS: The subjects described IBS as a chronic episodic illness that affects their daily lives. Interaction with the medical community seldom clarified understanding of the condition or improved its management. Three overriding themes emerged from the groups: a sense of frustration, a sense of isolation, and a search for a niche in the health/sick role continuum. Frustration was evident in the perceived inability to control symptoms, prevent episodes, identify episode triggers, and obtain medical validation of the condition. The constant anticipation of the next IBS episode, the need for immediate access to toilet facilities, and the nature of the bowel symptoms often required withdrawal from social activities and resultant isolation. CONCLUSIONS: IBS is perceived as a chronic condition resulting in frustration and social isolation, and physicians are perceived to be providing inadequate medical information or support to patients with IBS. PMID- 11401740 TI - Improving care for depression in organized health care systems. PMID- 11401741 TI - Treating depression in primary care practice: applications of research findings. PMID- 11401742 TI - Evaluation of the patient with dyspepsia. PMID- 11401743 TI - What is the clinical utility of obtaining a folate level in patients with macrocytosis or anemia? PMID- 11401744 TI - Can a clinical decision aid be useful in determining when to discontinue in hospital cardiac resuscitation? PMID- 11401745 TI - Does hormone replacement therapy (HRT) improve cognitive function or either delay or prevent dementia in postmenopausal women? PMID- 11401746 TI - Can 2 screening questions accurately detect alcohol and other drug abuse in the primary care setting? PMID- 11401747 TI - Multiple-problem patient visits. PMID- 11401748 TI - A new suicide. PMID- 11401749 TI - The silver lining. PMID- 11401751 TI - Improving depression care: barriers, solutions, and research needs. AB - Potential solutions for barriers to improved organization of care of depressive illness were identified. These included (1) aligning efforts to improve depression care with broader strategies for improving care of other chronic conditions; (2) increasing the availability of depression case management services in primary care; (3) developing registries and reminder systems to ensure active follow-up of depressed patients; (4) achieving agreement on how depression outcomes should be measured to provide outcomes-based performance standards; (5) providing greater support from mental health specialists for management of depressed patients by primary care providers; (6) campaigns to reduce the stigma associated with treatment of depressive illness; (7) increased dissemination of interventions that activate and empower patients managing a depressive illness; (8) redefining the lack of time of primary care providers for high-quality depression care as issues in organization of care and provider training; and (9) development of incentives (organizational or financial) for high-quality depression care. Research needs were identified according to what has been learned to date. Identified research needs included: studies of approaches to organization of case management, research in new populations (e.g., new diagnostic groups, rural populations, the disadvantaged, the elderly, and those with chronic medical illnesses), research on stepped care and relapse prevention strategies, evaluation of the societal benefits of improved depression care, and multisite trials and meta-analytic approaches that can provide adequate statistical power to assess societal benefits of improved care. PMID- 11401752 TI - Treating depressive disorders: who responds, who does not respond, and who do we need to study? AB - Numerous studies show that pharmacotherapy and psychotherapy are efficacious and effective in treating depression. Even though that is true for many people, there are some who do not respond to these therapies, whether it is because they possess characteristics that make them resistant to treatment or because the treatment is not acceptable to them. Also, there are other populations for whom research has yet to determine if guideline-level treatment is effective. People with comorbid Axis I disorders, certain minority groups, and children have been under-represented in treatment research. Future research should focus on treatment-resistant depression and special populations that have been understudied. PMID- 11401753 TI - Is peroxisome proliferation an obligatory precursor step in the carcinogenicity of di(2-ethylhexyl)phthalate (DEHP)? AB - Di(2-ethylhexyl)phthalate (DEHP), a peroxisome proliferator, has been listed by the International Agency for Research on Cancer (IARC) and by the National Toxicology Program as a possible or reasonably anticipated human carcinogen because it induces dose-related increases in liver tumors in both sexes of rats and mice. Recently, the suggestion has been advanced that DEHP should be considered unlikely to be a human carcinogen because it is claimed that the carcinogenic effects of this agent in rodents are due to peroxisome proliferation and that humans are nonresponsive to this process. An IARC working group recently downgraded DEHP to "not classifiable as to its carcinogenicity to humans" because they concluded that DEHP produces liver tumors in rats and mice by a mechanism involving peroxisome proliferation, which they considered to be not relevant to humans. The literature review presented in this commentary reveals that, although our knowledge of the mechanism of peroxisome proliferation has advanced greatly over the past 10 years, our understanding of the mechanism(s) of carcinogenicty of peroxisome proliferators remains incomplete. Most important is that published studies have not established peroxisome proliferation per se as an obligatory pathway in the carcinogenicity of DEHP. No epidemiologic studies have been reported on the potential carcinogenicity of DEHP, and cancer epidemiologic studies of hypolipidemic fibrate drugs (peroxisome proliferators) are inconclusive. Most of the pleiotropic effects of peroxisome proliferators are mediated by the peroxisome proliferator activated receptor (PPAR), a ligand activated transcription factor that is expressed at lower levels in humans than in rats and mice. In spite of this species difference in PPAR expression, hypolipidemic fibrates have been shown to induce hypolipidemia in humans and to modulate gene expression (e.g., genes regulating lipid homeostasis) in human hepatocytes by PPAR activation. Thus, humans are responsive to agents that induce peroxisome proliferation in rats and mice. Because peroxisome proliferators can affect multiple signaling pathways by transcriptional activation of PPAR regulated genes, it is likely that alterations in specific regulated pathways (e.g., suppression of apoptosis, protooncogene expression) are involved in tumor induction by peroxisome proliferators. In addition, because DEHP also induces biological effects that occur independently of peroxisome proliferation (e.g., morphologic cell transformation and decreased levels of gap junction intercellular communication), it is possible that some of these responses also contribute to the carcinogenicity of this chemical. Last, species differences in tissue expression of PPARs indicate that it may not be appropriate to expect exact site correspondence for potential PPAR-mediated effects induced by peroxisome proliferators in animals and humans. Because peroxisome proliferation has not been established as an obligatory step in the carcinogenicity of DEHP, the contention that DEHP poses no carcinogenic risk to humans because of species differences in peroxisome proliferation should be viewed as an unvalidated hypothesis. PMID- 11401754 TI - Evolutionary biology of plant defenses against herbivory and their predictive implications for endocrine disruptor susceptibility in vertebrates. AB - Hormone disruption is a major, underappreciated component of the plant chemical arsenal, and the historical coevolution between hormone-disrupting plants and herbivores will have both increased the susceptibility of carnivores and diversified the sensitivities of herbivores to man-made endocrine disruptors. Here I review diverse evidence of the influence of plant secondary compounds on vertebrate reproduction, including human reproduction. Three of the testable hypotheses about the evolutionary responses of vertebrate herbivores to hormone disrupting challenges from their diet are developed. Specifically, the hypotheses are that a) vertebrate herbivores will express steroid hormone receptors in the buccal cavity and/or the vomeronasal organ; b) absolute sex steroid concentrations will be lower in carnivores than in herbivores; and c) herbivore steroid receptors should be more diverse in their binding affinities than carnivore lineages. The argument developed in this review, if empirically validated by support for the specific hypotheses, suggests that a) carnivores will be more susceptible than herbivores to endocrine-disrupting compounds of anthropogenic origin entering their bodies, and b) diverse herbivore lineages will be variably susceptible to any given natural or synthetic contaminant. As screening methods for hormone-disrupting potential are compared and adopted, comparative endocrine physiology research is urgently needed to develop models that predict the broad applicability of those screening results in diverse vertebrate species. PMID- 11401755 TI - Gold-mining activities and mercury contamination of native amerindian communities in French Guiana: key role of fish in dietary uptake. AB - In 1994, the French National Public Health Network reported significant mercury exposure of native Amerindians in French Guiana. In 1997, a study was conducted in the Wayana community to quantify the dietary intake and to identify the fish species contributing the most to the contamination. The study was completed by an impregnation analysis based on Hg determination in hair samples. The methodology used was a detailed familial dietary study associated with Hg measurements in fish and some game. The study was conducted over 7 days in two different seasons in the four most populated Wayana villages on the upper part of the Maroni River (521 people; 70% of the Wayana population in French Guiana). Analysis was based on data on consumption obtained from 165 people in a 1-14 day period (i.e., 940 persons [times] days) and involved 270 fish samples from 48 species. Total Hg and monomethylmercury (MMHg) were also determined in hair samples (235 samples for total Hg). The results confirm mercury exposure of the Wayana population related to a diet rich in fish, which are relatively highly contaminated for certain species (up to 1.62 mg/kg fresh weight or 8.1 mg/kg dry weight in skeletal muscle). Results from hair samples showed that 57% of the Amerindians had Hg levels above the World Health Organization (WHO) safety limit (10 microg/g); all those over 1 year of age had a Hg intake greater than the WHO safety limit (200 microg MMHg/week for a 60-kg male). Hg concentrations in fish muscle were closely linked to the feeding regime and position of fish in the food webs. Overall, 14.5% of the fish collected exceeded the 0.5 mg/kg (fresh weight) safety limit. Four carnivorous species accounted for no less than 72% of the metal ingested by the Wayana families, although these represented only 28% of the consumed fish biomass. In conclusion, this study revealed excessive exposure to mercury in the Wayana population in French Guiana related to the consumption of contaminated fish. PMID- 11401756 TI - Identification of a P2X7 receptor in GH(4)C(1) rat pituitary cells: a potential target for a bioactive substance produced by Pfiesteria piscicida. AB - We examined the pharmacologic activity of a putative toxin (pPfTx) produced by Pfiesteria piscicida by characterizing the signaling pathways that induce the c fos luciferase construct in GH(4)C(1) rat pituitary cells. Adenosine-5' triphosphate (ATP) was determined to increase and, at higher concentrations, decrease luciferase activity in GH(4)C(1) rat pituitary cells that stably express c-fos luciferase. The inhibition of luciferase results from cytotoxicity, characteristic of the putative P. piscicida toxin (pPfTx). The actions of both pPfTx and ATP to induce c-fos luciferase were inhibited by the purinogenic receptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS). Further characterization of a P2X receptor on the GH(4)C(1) cell was determined by the analog selectivity of P2X agonists. The P2X1/P2X3 agonist alpha,beta methylene ATP (alpha,beta-MeATP) failed to increase or decrease c-fos luciferase. However, the P2X7 agonist 2',3'-(4-benzoyl)benzoyl ATP (BzATP), which had a predominant cytotoxic effect, was more potent than ATP. Immunoblot analysis of GH(4)C(1) cell membranes confirmed the presence of a 70-kDa protein that was immunoreactive to an antibody directed against the carboxy-terminal domain unique to the P2X7 receptor. The P2X7 irreversible antagonist oxidized-ATP (oxATP) inhibited the action of ATP, BzATP, and pPfTx. These findings indicate that GH(4)C(1) cells express purinogenic receptors with selectivity consistent with the P2X7 subtype and that this receptor pathway mediates the induction of the c fos luciferase reporter gene by ATP and the putative Pfiesteria toxin PMID- 11401757 TI - The impact of heat waves and cold spells on mortality rates in the Dutch population. AB - We conducted the study described in this paper to investigate the impact of ambient temperature on mortality in the Netherlands during 1979-1997, the impact of heat waves and cold spells on mortality in particular, and the possibility of any heat wave- or cold spell-induced forward displacement of mortality. We found a V-like relationship between mortality and temperature, with an optimum temperature value (e.g., average temperature with lowest mortality rate) of 16.5 degrees C for total mortality, cardiovascular mortality, respiratory mortality, and mortality among those [Greater and equal to] 65 year of age. For mortality due to malignant neoplasms and mortality in the youngest age group, the optimum temperatures were 15.5 degrees C and 14.5 degrees C, respectively. For temperatures above the optimum, mortality increased by 0.47, 1.86, 12.82, and 2.72% for malignant neoplasms, cardiovascular disease, respiratory diseases, and total mortality, respectively, for each degree Celsius increase above the optimum in the preceding month. For temperatures below the optimum, mortality increased 0.22, 1.69, 5.15, and 1.37%, respectively, for each degree Celsius decrease below the optimum in the preceding month. Mortality increased significantly during all of the heat waves studied, and the elderly were most effected by extreme heat. The heat waves led to increases in mortality due to all of the selected causes, especially respiratory mortality. Average total excess mortality during the heat waves studied was 12.1%, or 39.8 deaths/day. The average excess mortality during the cold spells was 12.8% or 46.6 deaths/day, which was mostly attributable to the increase in cardiovascular mortality and mortality among the elderly. The results concerning the forward displacement of deaths due to heat waves were not conclusive. We found no cold-induced forward displacement of deaths. PMID- 11401758 TI - A rat mammary tumor model induced by the organophosphorous pesticides parathion and malathion, possibly through acetylcholinesterase inhibition. AB - Environmental chemicals may be involved in the etiology of breast cancers. Many studies have addressed the association between cancer in humans and agricultural pesticide exposure. Organophosphorous pesticides have been used extensively to control mosquito plagues. Parathion and malathion are organophosphorous pesticides extensively used to control a wide range of sucking and chewing pests of field crops, fruits, and vegetables. They have many structural similarities with naturally occurring compounds, and their primary target of action in insects is the nervous system; they inhibit the release of the enzyme acetylcholinesterase at the synaptic junction. Eserine, parathion, and malathion are cholinesterase inhibitors responsible for the hydrolysis of body choline esters, including acetylcholine at cholinergic synapses. Atropine, a parasympatholytic alkaloid, is used as an antidote to acetylcholinesterase inhibitors. The aim of this study was to examine whether pesticides were able to induce malignant transformation of the rat mammary gland and to determine whether alterations induced by these substances increase the cholinergic activation influencing such transformation. These results showed that eserine, parathion, and malathion increased cell proliferation of terminal end buds of the 44-day-old mammary gland of rats, followed by formation of 8.6, 14.3, and 24.3% of mammary carcinomas, respectively, after about 28 months. At the same time, acetylcholinesterase activity decreased in the serum of these animals from 9.78 +/- 0.78 U/mL in the control animals to 3.05 +/- 0.06 U/mL; 2.57 +/- 0.15 U/mL; and 3.88 +/- 0.44 U/mL in the eserine-, parathion-, and malathion-treated groups, respectively. However, atropine alone induced a significant (p < 0.05) decrease in the acetylcholinesterase activity from the control value of 9.78 +/- 0.78 to 4.38 +/- 0.10 for atropine alone, to 1.32 +/- 0.06 for atropine in combination with eserine, and 2.39 +/- 0.29 for atropine with malathion, and there was no mammary tumor formation. These results indicate that organophosphorous pesticides induce changes in the epithelium of mammary gland influencing the process of carcinogenesis, and such alterations occur at the level of nervous system by increasing the cholinergic stimulation. PMID- 11401759 TI - Quantifying the effects of exposure to indoor air pollution from biomass combustion on acute respiratory infections in developing countries. AB - Acute respiratory infections (ARI) are the leading cause of burden of disease worldwide and have been causally linked with exposure to pollutants from domestic biomass fuels in developing countries. We used longitudinal health data coupled with detailed monitoring and estimation of personal exposure from more than 2 years of field measurements in rural Kenya to estimate the exposure-response relationship for particulates < 10 microm diameter (PM(10)) generated from biomass combustion. Acute respiratory infections and acute lower respiratory infections are concave, increasing functions of average daily exposure to PM(10), with the rate of increase declining for exposures above approximately 1,000-2,000 microg/m(3). This first estimation of the exposure-response relationship for the high-exposure levels characteristic of developing countries has immediate and important consequences for international public health policies, energy and combustion research, and technology transfer efforts that affect more than 2 billion people worldwide. PMID- 11401760 TI - Coupling between annual and ENSO timescales in the malaria-climate association in Colombia. AB - We present evidence that the El Nino phenomenon intensifies the annual cycle of malaria cases for Plasmodium vivax and Plasmodium falciparum in endemic areas of Colombia as a consequence of concomitant anomalies in the normal annual cycle of temperature and precipitation. We used simultaneous analyses of both variables at both timescales, as well as correlation and power spectral analyses of detailed spatial (municipal) and temporal (monthly) records. During "normal years," endemic malaria in rural Colombia exhibits a clear-cut "normal" annual cycle, which is tightly associated with prevalent climatic conditions, mainly mean temperature, precipitation, dew point, and river discharges. During historical El Nino events (interannual time scale), the timing of malaria outbreaks does not change from the annual cycle, but the number of cases intensifies. Such anomalies are associated with a consistent pattern of hydrological and climatic anomalies: increase in mean temperature, decrease in precipitation, increase in dew point, and decrease in river discharges, all of which favor malaria transmission. Such coupling explains why the effect appears stronger and more persistent during the second half of El Nino's year (0), and during the first half of the year (+1). We illustrate this finding with data for diverse localities in Buenaventura (on the Pacific coast) and Caucasia (along the Cauca river floodplain), but conclusions have been found valid for multiple localities throughout endemic regions of Colombia. The identified coupling between annual and interannual timescales in the climate-malaria system shed new light toward understanding the exact linkages between environmental, entomological, and epidemiological factors conductive to malaria outbreaks, and also imposes the coupling of those timescales in public health intervention programs. PMID- 11401761 TI - Biomarker correlations of urinary 2,4-D levels in foresters: genomic instability and endocrine disruption. AB - Forest pesticide applicators constitute a unique pesticide use group. Aerial, mechanical-ground, and focal weed control by application of herbicides, in particular chlorophenoxy herbicides, yield diverse exposure scenarios. In the present work, we analyzed aberrations in G-banded chromosomes, reproductive hormone levels, and polymerase chain reaction-based V(D)J rearrangement frequencies in applicators whose exposures were mostly limited to chlorophenoxy herbicides. Data from appliers where chlorophenoxy use was less frequent were also examined. The biomarker outcome data were compared to urinary levels of 2,4 dichlorophenoxyacetic acid (2,4-D) obtained at the time of maximum 2,4-D use. Further comparisons of outcome data were made to the total volume of herbicides applied during the entire pesticide-use season.Twenty-four applicators and 15 minimally exposed foresters (control) subjects were studied. Categorized by applicator method, men who used a hand-held, backpack sprayer in their applications showed the highest average level (453.6 ppb) of 2,4-D in urine. Serum luteinizing hormone (LH) values were correlated with urinary 2,4-D levels, but follicle-stimulating hormone and free and total testosterone were not. At the height of the application season; 6/7 backpack sprayers, 3/4 applicators who used multinozzle mechanical (boom) sprayers, 4/8 aerial applicators, and 2/5 skidder radiarc (closed cab) appliers had two or more V(D)J region rearrangements per microgram of DNA. Only 5 of 15 minimally exposed (control) foresters had two or more rearrangements, and 3 of these 5 subjects demonstrated detectable levels of 2,4-D in the urine. Only 8/24 DNA samples obtained from the exposed group 10 months or more after their last chlorophenoxy use had two rearrangements per microgram of DNA, suggesting that the exposure-related effects observed were reversible and temporary. Although urinary 2,4-D levels were not correlated with chromosome aberration frequency, chromosome aberration frequencies were correlated with the total volume of herbicides applied, including products other than 2,4-D. In summary, herbicide applicators with high urinary levels of 2,4-D (backpack and boom spray applications) exhibited elevated LH levels. They also exhibited altered genomic stability as measured by V(D)J rearrangement frequency, which appears reversible months after peak exposure. Though highly detailed, the limited sample size warrants cautious interpretation of the data. PMID- 11401762 TI - Examination of the melatonin hypothesis in women exposed at night to EMF or bright light. AB - It has been hypothesized that the increased incidence of breast cancer in industrial societies is related to greater exposure to power-frequency electric and magnetic fields (EMF) and/or the presence of high levels of light at night (LAN). EMF and LAN are said to reduce circulating levels of the hormone melatonin which, in turn, allows estrogen levels to rise and stimulate the turnover of breast epithelial stem cells and increase the risk for malignant transformation. Three laboratory-based studies, in which a total of 53 healthy young women were exposed at night to EMF or to LAN under controlled exposure conditions, were performed to determine whether such exposures reduce melatonin and are associated with further alterations in estrogen. All-night exposure to industrial-strength magnetic fields (60 Hz, 28.3 microT) had no effect on the blood levels of melatonin or estradiol. In contrast, nocturnal melatonin levels were profoundly suppressed, and the time of peak concentration was significantly delayed in women exposed to LAN, regardless of whether they were in the follicular or luteal phase of the menstrual cycle. These changes, however, were not associated with alterations in point-for-point matching measures of estradiol. Women who chronically secrete high or low amounts of melatonin each night (area-under-curve range: 86-1,296 pg/mL) also did not differ in their blood levels of estradiol. Taken together, these results are consistent with a growing body of evidence which generally suggests that environmental EMF exposure has little or no effect on the parameters measured in this report. PMID- 11401763 TI - Data quality in predictive toxicology: reproducibility of rodent carcinogenicity experiments. AB - We compared 121 replicate rodent carcinogenicity assays from the two parts (National Cancer Institute/National Toxicology Program and literature) of the Carcinogenic Potency Database (CPDB) to estimate the reliability of these experiments. We estimated a concordance of 57% between the overall rodent carcinogenicity classifications from both sources. This value did not improve substantially when additional biologic information (species, sex, strain, target organs) was considered. These results indicate that rodent carcinogenicity assays are much less reproducible than previously expected, an effect that should be considered in the development of structure-activity relationship models and the risk assessment process. PMID- 11401764 TI - Diesel exhaust particles suppress macrophage function and slow the pulmonary clearance of Listeria monocytogenes in rats. AB - In this study, we tested the hypothesis that exposure to diesel exhaust particles (DEP) may increase susceptibility of the host to pulmonary infection. Male Sprague-Dawley rats received a single dose of DEP (5 mg/kg), carbon black (CB, 5 mg/kg), or saline intratracheally. Three days later, the rats were inoculated intratracheally with approximately 5,000 Listeria monocytogenes and sacrificed at 3, 5, and 7 days postinfection, and we determined the number of viable Listeria in the left lobe of lungs. The remaining lungs underwent bronchoalveolar lavage (BAL) and the retrieved BAL cells were identified and counted. Luminol-dependent chemiluminescence, a measure of reactive oxygen species (ROS) formation, generated by BAL cells was monitored and the levels of nitric oxide and tumor necrosis factor (TNF)-[alpha] produced by macrophages in culture were determined. At 7 days postinfection, we excised the lung-draining lymph nodes and phenotyped the lymphocyte subpopulations. Exposure of rats to DEP, but not to CB, decreased the clearance of Listeria from the lungs. Listeria-induced generation of luminol dependent chemiluminescence by pulmonary phagocytes decreased by exposure to DEP but not CB. Similarly, Listeria-induced production of NO by alveolar macrophages was negated at 3, 5, and 7 days after inoculation in DEP-exposed rats. In contrast, CB exposure had no effect on Listeria-induced NO production at 3 days after infection and had a substantially smaller effect than DEP at later days. Exposure to DEP or CB resulted in enlarged lung-draining lymph nodes and increased the number and percentage of CD4(+) and CD8(+) T cells. These results showed that exposure to DEP decreased the ability of macrophages to produce antimicrobial oxidants in response to Listeria, which may play a role in the increased susceptibility of rats to pulmonary infection. This DEP-induced suppression is caused partially by chemicals adsorbed onto the carbon core of DEP, because impaired macrophage function and decreased Listeria clearance were not observed following exposure to CB. PMID- 11401765 TI - Formamidopyrimidine-DNA glycosylase enhances arsenic-induced DNA strand breaks in PHA-stimulated and unstimulated human lymphocytes. AB - To confirm that arsenic (As) induces oxidative DNA damage in phytohemagglutinin (PHA)-stimulated and unstimulated human lymphocytes, we used the alkaline comet assay combined with specific enzyme [formamidopyrimidine-DNA glycosylase (FPG)] digestion to measure As-induced base damage. The results showed that the enzyme sensitive sites were readily detected with the alkaline comet assay after the cells were treated with 10 microM As for 2 hr. The repair patterns observed for FPG-created DNA single strand breaks (SSBs) in As-treated cells were comparable to those in hydrogen peroxide (H(2)O(2))-treated cells. The enzyme-created SSBs, As-induced base damage, were more significant in PHA-stimulated lymphocytes. About 63% and 68% of SSBs induced by As and H(2)O(2), respectively, were repaired in PHA-stimulated lymphocytes by 2-hr repair incubation, but about 34% and 43%, respectively, were repaired in unstimulated cells. About 40% and 49% of base damage induced by As and H(2)O(2), respectively, were repaired in PHA-stimulated lymphocytes, but about 19% and 21%, respectively, were repaired in unstimulated cells. These results indicated that As induced oxidative DNA damage in human lymphocytes at micromolar concentrations. The damaged bases could be chiefly purines or formamidopyrimidines. Like the damage induced by H(2)O(2), As-induced DNA damage was repaired more slowly in unstimulated lymphocytes. PMID- 11401766 TI - Interrelations of lead levels in bone, venous blood, and umbilical cord blood with exogenous lead exposure through maternal plasma lead in peripartum women. AB - Recent research has raised the possibility that fetal lead exposure is not estimated adequately by measuring lead content in maternal whole blood lead because of the variable partitioning of lead in whole blood between plasma and red blood cells. Lead in maternal plasma may derive in large part from maternal bone lead stores. In this study we aimed to estimate the contribution of maternal whole blood lead, maternal bone lead levels, and environmental lead to umbilical cord blood lead levels (as a measure of fetal lead exposure). In the model, we assumed that lead from all of these sources reaches the fetus through the maternal plasma lead pathway. In 1994-1995, we recruited 615 pregnant women for a study of lead exposure and reproductive outcomes in Mexico City. We gathered maternal and umbilical cord blood samples within 12 hr of each infant's delivery and measured maternal lead levels in cortical bone and trabecular bone by a K-X ray fluorescence (K-XRF) instrument within 1 month after delivery. We administered a questionnaire to assess use of lead-glazed ceramics (LGC) to cook food and we obtained data on regional air lead levels during the 2 months before delivery. We used structural equation models (SEMs) to estimate plasma lead as the unmeasured (latent) variable and to quantify the interrelations of plasma lead, the other lead biomarkers, and environmental lead exposure. In the SEM analysis, a model that allowed plasma lead to vary freely from whole blood lead explained the variance of cord blood lead (as reflected by a total model R(2); R(2) = 0.79) better than did a model without plasma lead (r(2) = 0.67). Cortical bone lead, trabecular bone lead, use of LGC, and mean air lead level contributed significantly to plasma lead. The exchange of lead between plasma and red blood cells was mostly in the direction of plasma to cells. According to the final model, an increase in trabecular bone lead and cortical bone lead was associated with increases in cord blood lead of 0.65 and 0.25 microg/dL, respectively. An increase of 0.1 microg/m(3) in air lead was associated with an increase in the mean level of fetal cord blood lead by 0.67 microg/dL. With one additional day of LCG use per week in the peripartum period, the mean fetal blood lead level increased by 0.27 microg/dL. Our analyses suggested that maternal plasma lead varies independently from maternal whole blood lead and that the greatest influences on maternal plasma lead are maternal bone lead stores, air lead exposures, and recent cooking with LGC. The contributions from endogenous (bone) and exogenous (environmental) sources were relatively equal. Measurement of plasma and bone lead may be important in accurately assessing fetal lead exposure and its major sources, particularly if exogenous exposures decline. PMID- 11401767 TI - Work characteristics and pesticide exposures among migrant agricultural families: a community-based research approach. AB - There are few data on pesticide exposures of migrant Latino farmworker children, and access to this vulnerable population is often difficult. In this paper we describe a community-based approach to implement culturally appropriate research methods with a migrant Latino farmworker community in Oregon. Assessments were conducted in 96 farmworker homes and 24 grower homes in two agricultural communities in Oregon. Measurements included surveys of pesticide use and work protection practices and analyses of home-dust samples for pesticide residues of major organophosphates used in area crops. Results indicate that migrant farmworker housing is diverse, and the amounts and types of pesticide residues found in homes differ. Azinphos-methyl (AZM) was the pesticide residue found most often in both farmworker and grower homes. The median level of AZM in farmworker homes was 1.45 ppm compared to 1.64 ppm in the entry area of grower homes. The median level of AZM in the play areas of grower homes was 0.71 ppm. The levels of AZM in migrant farmworker homes were most associated with the distance from fields and the number of agricultural workers in the home. Although the levels of AZM in growers and farmworker homes were comparable in certain areas, potential for disproportionate exposures occur in areas of the homes where children are most likely to play. The relationship between home resident density, levels of pesticide residues, and play behaviors of children merit further attention. PMID- 11401768 TI - Possible estuary-associated syndrome: symptoms, vision, and treatment. AB - The human illness designated as possible estuarine-associated syndrome (PEAS) by the Centers for Disease Control and Prevention (CDC) has been associated with exposure to estuaries inhabited by toxin-forming dinoflagellates, including members of the fish-killing toxic Pfiesteria complex (TPC), Pfiesteria piscicida and Pfiesteria shumwayae. Humans may be exposed through direct contact with estuarine water or by inhalation of aerosolized or volatilized toxin(s). The five cases reported here demonstrate the full spectrum of symptoms experienced during acute and chronic stages of this suspected neurotoxin-mediated illness. The nonspecific symptoms most commonly reported are cough, secretory diarrhea, headache, fatigue, memory impairment, rash, difficulty in concentrating, light sensitivity, burning skin upon water contact, muscle ache, and abdominal pain. Less frequently encountered symptoms are upper airway obstruction, shortness of breath, confusion, red or tearing eyes, weakness, and vertigo. Some patients experience as few as four of these symptoms. The discovery that an indicator of visual pattern-detection ability, visual contrast sensitivity (VCS), is sharply reduced in affected individuals has provided an objective indicator that is useful in diagnosing and monitoring PEAS. VCS deficits are present in both acute and chronic PEAS, and VCS recovers during cholestyramine treatment coincident with symptom abatement. Although PEAS cannot yet be definitively associated with TPC exposure, resolution with cholestyramine treatment suggests a neurotoxin mediated illness. PMID- 11401769 TI - Toxic beryllium and genetic testing. PMID- 11401770 TI - Cellular telephones and brain cancer: current research. PMID- 11401772 TI - Baby ills from beauty aids? PMID- 11401773 TI - CDC unveils body burden. PMID- 11401774 TI - Boston pee party. PMID- 11401775 TI - EC says shhh! PMID- 11401776 TI - Health oasis in the desert southwest. PMID- 11401777 TI - Ancient teachings, modern lessons. PMID- 11401778 TI - Indi-gene-ous conflict. PMID- 11401779 TI - Sound science: ferries keep watch on coastal waters. PMID- 11401780 TI - Embryonic genome activation. AB - Genome activation is one of the first critical events in the life of the new organism. Both the timing of genome activation and the array of genes activated must be controlled correctly. Genome activation occurs in a stepwise manner, with some genes being transcribed well in advance of the major genome activation event, in which most housekeeping genes become activated. Changes in chromatin protein content, particularly histone proteins, and chromatin structure appear to regulate the availability of the genome for transcription and provide for specificity of transcription. Gene enhancers are not initially required for transcription, but become necessary as the chromatin structure is modified. Changes in transcription factor content or activity are also required, and protein synthesis is essential for genome activation during both early and later phases of transcriptional activation. Both the changes in chromatin structure and availability of transcription factors are regulated by cell cycle-dependent mechanisms, thus providing the necessary coordination between these processes and other processes such as DNA replication and cleavage. PMID- 11401781 TI - Bioimmunotherapeutic targets on angiogenetic blood vessels in solid malignangies. AB - Physiological angiogenesis is a tightly regulated process that occurs mainly during reproduction, development and wound healing. Although angiogenesis is a continuous process, different consecutive steps can be identified, including: i) release of pro-angiogenetic factors; ii) release of proteolytic enzymes; iii) endothelial cell migration, morphogenesis and proliferation. Angiogenesis is also a hallmark of malignant diseases, and an inverse correlation between tumor vascularity and survival was demonstrated. Thus, strategies aimed at interfering with tumor blood supply by targeting tumor vasculature, presently represent promising new approaches for the treatment of solid malignancies. In fact, at least 30 angiogenetic inhibitors, utilized alone or in combination with other therapeutic agents, are currently being tested in clinical trials in humans. In this paper, we will review current knowledges on selected molecules expressed by endothelial cells and involved in distinct steps of the angiogenetic process, that represent potential targets for bioimmunotherapeutic approaches in human malignancies. PMID- 11401782 TI - Designer cancer vaccines made easy: protein transfer of immunostimulatory molecules for use in therapeutic tumor vaccines. AB - Advances in the understanding of the immune response to tumors has led to the development of new strategies to design therapeutic vaccines. One of these strategies is the development of protein transfer of immunostimulatory molecules onto the surface of tumor cells, thereby directing the immune response to the tumor antigens carried by the modified tumor cells. This strategy has been developed as an alternative to gene transfer, the more classical technique of introducing immunostimulatory molecules onto tumors. In this report we briefly review current strategies for immunotherapy and then focus on several approaches to protein transfer and their historical basis. Finally, the application of these protein transfer approaches to develop cancer vaccines are reviewed and discussed. PMID- 11401783 TI - Entry mechanisms of mycobacteria. AB - Since many mycobacteria are facultative intracellular pathogens, their ability to cause disease involves entry, survival and replication within host cells. Despite the fact that mycobacteria were first associated with disease more than 125 years ago, the first step in the production of an infection, entry into host cells, is not well understood. Mycobacteria have the ability to enter a number of different cell types, but the primary cell type that they are thought to replicate within during human disease is macrophages. Since macrophages have a large number of receptors that are designed for relatively non-specific uptake of foreign particles, there are multiple routes by which nearly any bacteria can be taken up. The outcome of mycobacterial entry into macrophages via different mechanisms is unclear. Although it is thought that mycobacteria may enter macrophages by a mechanism that allows them to avoid lysosomal fusion, it remains possible that mycobacteria enter by more than one mechanism, yet remain viable and replicate intracellularly through modification of the phagosome. In the current discussion we will review mycobacterial research specifically relating to the mechanisms of entry into host cells. Although much progress has been made in our understanding of entry by mycobacteria, we anticipate that clarification of the role of entry in pathogenesis will require further application of newly developed molecular tools to dissect each of the proposed mechanisms. PMID- 11401784 TI - Study of the mechanism underlying the difference in motility between the large and small intestine: the "single" and "multiple" pacemaker theory. AB - The common movement of the small intestine (SI) is peristalsis and of the large intestine (LI) are giant migrating contractions. The mechanism underlying the difference in the type of motility between the SI and LI is yet to be elucidated; the current communication investigated this point. 7 pigs (4 male) were anesthetized, abdomen opened and a balloon-ended catheter was introduced into the right colon through ileotomy. Balloon was filled in increments of 10 ml of saline and the EMG activity of the colonic longitudinal and circular muscle layers was recorded before and after myotomy performed between 2 of the 3 electrodes fixed to the colon. The balloon was then withdrawn, located in the terminal ileum and distended in increments of 2 ml; the ileal EMG activity was registered before and after myotomy done between 2 of the 3 electrodes applied to the ileum. The LI showed slow waves or pacesetter potentials (PPs) and action potentials (APs) which had the same frequency, amplitude and conduction velocity from the 3 electrodes of the same animal. The waves were recorded from the longitudinal and not the circular muscle coat. Upon LI distension, the electric activity increased and was recorded also from the circular muscle. At 40-50 ml distension, the balloon was dispelled to the transverse colon. Electric activity from SI was similar to that of the LI, but was not the same from the 3 electrodes; it diminished aborally. It increased with increasing balloon distension until, at 8 10 ml distension, the balloon moved slowly aborally. Electric waves were recorded proximally but not distally to colonic myotomy, and the balloon moved up to the cut. In the SI, waves were recorded both proximally and distally to the ileal myotomy, and the balloon moved across the cut. The fact that the colonic electric waves displayed the same variables from the 3 electrodes and that they were not recorded distally to the colonic myotomy, would suggest the presence of a "single" colonic pacemaker, probably situated in the cecum. This is in contrast to the hypothesis of the "multiplicity" of the small intestinal pacemakers, which is based on the fact that the electric activity diminished as the waves propagated aborally and on the existence of slow waves distal to the ileal myotomy. The concept of "single" and "multiple" pacemakers explaining the difference in the motility of small from that of large gut needs further studies. PMID- 11401785 TI - Management theory and applications. AB - Management is critical as an organization pursues its mission. There are many theories of management, but all agree that an effective organizational structure can facilitate the operation of a company. The author describes the typical functional areas found in most organizations (finance, operations, marketing, information systems, legal, and human resources); examines how the organization of tasks and people are inter-linked; and shows that administrators who have a working knowledge of management theory tend to be effective in the performance of their jobs. PMID- 11401786 TI - An overview of economics. AB - Economics influences health care at every level. Part of that influence is the result of the interaction of buyers and sellers in markets. Another part is the interaction among aggregates of different markets, the government, and international actors. This article provides a short, non-technical introduction to basic theories of both micro and macro economics as a necessary precursor to the understanding of more complex theories and debates about the economics of health care. PMID- 11401787 TI - Information systems. AB - Information is critical for the successful operation of healthcare facilities. Data must be accurate, available on a timely basis, and accessible. An information system should connect all potential suppliers and users of data; be accessible to users, but secure from unauthorized persons; have equipment that can interface with sources and users throughout an organization; archive data in a logical and accessible manner; and support users throughout the organization it serves. Dr. Fallon discusses operator training, strategic planning, and upgrading of equipment. PMID- 11401788 TI - An overview of accounting and budgeting. AB - Accounting is inextricably linked with daily professional practice and commerce. No individual, organization, or business can survive without some understanding of the basic concepts of accounting. Having knowledge of fundamental concepts of accounting will relieve some of the pressures imposed by governmental regulatory agencies and legislative requirements, and assist healthcare workers in operating more effectively and competing more successfully. PMID- 11401789 TI - Essentials of finance for occupational physicians. AB - Comprehending the principles of finance is paramount to understanding the way an organization chooses to generate and use its financial resources. Financial principles may be employed in the same way a physician reviews fundamental systems to gauge a person s health. Just as basic anatomical and physiological components are used to assess the health of an individual, basic financial elements exist to ascertain the health of an organization. This chapter explains risk assessment, accounts receivable management, inventory, depreciation, capital formation, ratio analysis, and more. PMID- 11401790 TI - Concepts of organizational behavior applied to occupational medicine. AB - People in organizational settings exhibit predictable patterns of behavior. Effective managers understand the psychological underpinnings of group functions. Formal organizational rules and job expectations influence employees. Informal or group dynamics also exert powerful influences. Effective managers try to reduce conflict between organizational and group norms. Informal group cohesion can be harnessed and channeled towards achieving organizational goals and objectives, and informal leaders can be assets to organizational managers. While formal organizational structures can be changed, supervisors should try to accommodate informal group structures when possible. Understanding subordinates is an important skill of successful managers. PMID- 11401791 TI - Marketing fundamentals. AB - This chapter outlines current marketing practice from a managerial perspective. The role of marketing within an organization is discussed in relation to efficiency and adaptation to changing environments. Fundamental terms and concepts are presented in an applied context. The implementation of marketing plans is organized around the four P's of marketing: product (or service), promotion (including advertising), place of delivery, and pricing. These are the tools with which marketers seek to better serve their clients and form the basis for competing with other organizations. Basic concepts of strategic relationship management are outlined. Lastly, alternate viewpoints on the role of advertising in healthcare markets are examined. PMID- 11401792 TI - Human resource management. AB - The occupational health physician can take advantage of the broad functions of human resource management to offer care and treatment in a very practical way in the day-to-day workplace. Understanding the capabilities and services of the human resource department can aid the occupational health practitioner in maintaining employee health and productivity. Thus, the individual is helped as well as the organization. This article describes the functions of the human resource department and delineates the close connection between it and the occupational health physician. PMID- 11401793 TI - Fundamentals of compensation and benefits. AB - The authors discuss the objectives of compensation programs--including direct and indirect forms of reward--and employee motivation. Job descriptions and job evaluations are also explored. PMID- 11401794 TI - The older worker. AB - About one person in eight remains employed past 65, the average age for retirement in the U.S. These persons tend to be highly reliable. They can adapt and learn new technology, but may require extra time to do so. Older workers have particular needs in the workplace due to physiological changes that accompany aging. They may require more lighting, and they may have decreased mobility, physical strength, and dexterity. These factors often have no impact on their ability to accomplish job duties. This chapter underscores the significant contributions that older workers often provide, and also addresses retirement planning. PMID- 11401795 TI - Administrative issues related to addiction in the workplace. AB - Drug abuse is an important problem in the workplace. Seventy percent of current illegal drug users are employed, and approximately 7% of Americans employed in full-time work report heavy drinking. Drug-using employees are twice as likely to request time off, and 3.6 times more likely to be involved in a workplace accident. Individuals who use alcohol or other drugs in the workplace annually cost American business 81 billion dollars in lost productivity; 86% of these costs are attributed to drinking. Although alcohol is the primary source of problems for the workplace, other drugs such as amphetamines, cocaine, marijuana, and tobacco are also troublesome. Employee assistance programs and drug treatment hold great promise for coping with these problematic substances. PMID- 11401796 TI - Ethics in the practice of occupational medicine. AB - Ethical considerations are often important when managers make organizational decisions. There are two ethical approaches commonly employed when making decisions. The first is deontology or the existence of an absolute standard that does not change over time. Decisions made by reference to a creed or professional code are uniformly consistent. The second ethical system is teleology or optimization. Decisions are made to optimize the outcome, and they can be inconsistent over time. Health professionals must often choose their allegiance from several alternatives. These conflicts must be recognized and reconciled. Students can benefit from the inclusion of ethical training during their professional preparation. PMID- 11401797 TI - Can special circumstances justify republication? A difference of opinions. PMID- 11401798 TI - Student-initiated forum on abortion. PMID- 11401800 TI - Confronting racial and ethnic disparities in health care. PMID- 11401801 TI - Innovative approaches to educating medical students for practice in a changing health care environment: the National UME-21 Project. AB - In today's continually changing health care environment, there is serious concern that medical students are not being adequately prepared to provide optimal health care in the system where they will eventually practice. To address this problem, the Health Resources and Services Administration (HRSA) developed a $7.6 million national demonstration project, Undergraduate Medical Education for the 21st Century (UME-21). This project funded 18 U.S. medical schools, both public and private, for a three-year period (1998-2001) to implement innovative educational strategies. To accomplish their goals, the 18 UME-21 schools worked with more than 50 organizations external to the medical school (e.g., managed care organizations, integrated health systems, Area Health Education Centers, community health centers). The authors describe the major curricular changes that have been implemented through the UME-21 project, discuss the challenges that occurred in carrying out those changes, and outline the strategies for evaluating the project. The participating schools have developed curricular changes that focus on the core primary care clinical clerkships, take place in ambulatory settings, include learning objectives and competencies identified as important to providing care in the future health care system, and have faculty development and internal evaluation components. Curricular changes implemented at the 18 schools include having students work directly with managed care organizations, as well as special demonstration projects to teach students the knowledge, skills, and attitudes necessary for successfully managing care. It is already clear that the UME-21 project has catalyzed important curricular changes within 12.5% of U.S. medical schools. The ongoing national evaluation of this project, which will be completed in 2002, will provide further information about the project's impact and effectiveness. PMID- 11401802 TI - Vanquishing virtue: the impact of medical education. AB - North American physicians emerge from their medical training with a wide array of professional beliefs and values. Many are thoughtful and introspective. Many are devoted to patients' welfare. Some bring to their work a broad view of social responsibility. Nonetheless, the authors contend that North American medical education favors an explicit commitment to traditional values of doctoring empathy, compassion, and altruism among them-and a tacit commitment to behaviors grounded in an ethic of detachment, self-interest, and objectivity. They further note that medical students and young physicians respond to this conflict in various ways. Some re-conceptualize themselves primarily as technicians and narrow their professional identities to an ethic of competence, thus adopting the tacit values and discarding the explicit professionalism. Others develop non reflective professionalism, an implicit avowal that they best care for their patients by treating them as objects of technical services (medical care). Another group appears to be "immunized" against the tacit values, and thus they internalize and develop professional virtue. Certain personal characteristics of the student, such as gender, belief system, and non-medical commitments, probably play roles in "immunization," as do medical school features such as family medicine, communication skills courses, medical ethics, humanities, and social issues in medicine. To be effective, though, these features must be prominent and tightly integrated into the medical school curriculum. The locus of change in the culture of medicine has now shifted to ambulatory settings and the marketplace. It remains to be seen whether this move will lessen the disjunction between the explicit curriculum and the manifestly contradictory values of detachment and entitlement, and the belief that the patient's interest always coincides with the physician's interest. PMID- 11401803 TI - A synthesis of nine major reports on physicians' competencies for the emerging practice environment. AB - Medical education and training programs generally have been slow to introduce curriculum content that reflects important changes in practice organization and health services delivery. However, impetus for curricular reform is gaining momentum as national organizations endorse new content for both medical school and residency education. The authors and colleagues at Tufts Managed Care Institute reviewed nine reports by key national organizations to assess their positions on curricular reform in light of changes in practice and the system of care. The reports agree generally on the evolving nature of practice, the need to address these changes during medical school and residency training, and the description of the new curriculum content that they advocate. The authors grouped these reports' specific recommendations under ten curriculum domains: health care system overview; population-based care; quality measurement and improvement; medical management; preventive care; physician-patient communication; ethics; teamwork and collaboration; information management and technology; and practice management. They describe the reports' rationales and cite specific knowledge and skills that these national organizations identify within each domain. This domain based framework synthesizes and complements the recommendations of these national organizations. The authors conclude that implementing curricular reform remains a challenge. The information and competencies need to be organized and sequenced for stage of training and specialty, and barriers to change require strategic and operational planning. Having a common nomenclature and framework will facilitate the introduction of new content within schools and programs, across departments, and among institutions nationwide. PMID- 11401804 TI - "Bench-to-bedside"--the wrong paradigm for patient-oriented investigation. PMID- 11401805 TI - Is the master clinician dead? PMID- 11401806 TI - The spirit catches you and you fall down: a Hmong child, her American doctors, and the collision of two cultures. PMID- 11401808 TI - Surveying graduates of one school to determine regional workforce demand. AB - PURPOSE: To study the demand for physician graduates from one school in one region of the country. The use of demand as a measure of potential regional variation should be of interest to medical educators and policymakers. METHOD: All residency graduates of the University of Washington School of Medicine between 1975 and 1995 (n = 3,824) were surveyed about their ability to gain employment in a timely manner and whether they were recruiting physicians for their practices. RESULTS: The response rate was 50.29%. A non-responder survey was done using a subsample (n = 200), with a 28% return. Over 95% of the graduates had found employment in their desired specialties and locations within two years of finishing their residencies. This was the same for graduates over all years. Approximately 30% of all practices of respondents within the Northwest region were recruiting for new physicians (26% of specialty practices and 34% of generalist practices were recruiting). There was no difference between recruitment in the urban and rural practices or between respondents to the initial survey and those responding to the follow up. CONCLUSIONS: Despite a significant oversupply of specialist physicians and at least a sufficient supply of generalist physicians nationally, there appears to be a strong demand for both specialists and generalists in the Northwest region of the country. This raises questions concerning the use of national averages to inform the education policies in specific regions of the country. More validated measures of demand are needed for future studies. PMID- 11401809 TI - Effects of perceptions and mentorship on pursuing a career in academic medicine in obstetrics and gynecology. AB - PURPOSE: To understand the perceptions of residents and Fellows in obstetrics and gynecology about the impacts of race or ethnicity, gender, and mentorship experiences on pursuing careers in academic medicine. METHOD: Two surveys were administered: one to a sample of 2,000 Fellows of the American College of Obstetricians and Gynecologists, and one to the 4,814 obstetrics and gynecology residents taking the 1998 in-training examination. The questionnaires asked about demographics, perceptions about careers in academic medicine, and residents' experiences with mentorship. RESULTS: Response rates were 96.8% for residents and 40.6% for FELLOWS: Of the residents, 26.1% indicated they would not consider a career in academic medicine. First-year women residents were more inclined to pursue careers in academic medicine than were first-year men (p =.042), but their interest declined during residency. Women residents (43%)-especially minorities felt that men were mentored and recruited more for faculty positions, while men (38%) felt that women were mentored and recruited more. Fellows' reports of recruitment did not differ by gender. Most white residents did not perceive racial or ethnic bias in mentoring or recruiting, while most non-white residents did. Almost one third of non-white women residents felt that supervisors were more likely to condescend to women and minority individuals. CONCLUSIONS: It is likely that neither men nor women residents in obstetrics and gynecology receive adequate mentorship for careers in academic medicine. Perceptions of bias are a serious barrier to developing racial, ethnic, and gender diversity in leadership positions. PMID- 11401810 TI - A pilot study assessing the influences of charge data and group process on diagnostic test ordering by residents. AB - PURPOSE: Providing charge data to resident physicians has been shown to reduce the amounts spent on diagnostic testing. This pilot study sought to determine the influences of charge data and group decision making on diagnostic test ordering by internal medicine residents. METHOD: In an interactive workshop, 23 internal medicine residents received a hypothetical case. They completed an 18-item questionnaire estimating charges for diagnostic tests and then "ordered" tests. The residents were then randomly divided into groups that either received charge data, received charge data after ordering tests, or received no charge data. The groups ordered tests by consensus. Tests were weighted for appropriateness (+1 to +6) and inappropriateness (-1 to -6). Analyses compared individual and group decisions and effect of availability of charge data. RESULTS: Residents with access to charge data spent less on tests, but also had lower appropriateness scores. The appropriateness of the diagnostic workup was better by groups than by individuals, but cost more. CONCLUSION: Cost-containment interventions targeted towards doctors in training need to address the effect on quality of care and the influence of the group process in clinical decision making. Group diagnostic decisions may be more costly, but more appropriate. PMID- 11401811 TI - Preliminary reports and the rates of publication of follow-up reports in peer reviewed, indexed journals. AB - PURPOSE: To test the hypothesis that articles published as "preliminary" or "pilot" reports are followed by more definitive publications in only a minority of cases. METHOD: A survey of Medline was performed for reports published in 1992 in journals listed in the Abridged Index Medicus that had the word "preliminary" or "pilot" in the title. For identified reports, a Medline search of publications in 1992 through 1999 was performed, using lead author's name, second author's name, and senior (last) author's name, and at least one keyword based on the publication title. Preliminary and pilot publications were subdivided by type of study (controlled clinical study, case series, laboratory or nonclinical) and by the report of either positive or negative results. Rates of publication based on study design and publication bias were compared using the chi-square test for statistical significance. RESULTS: The rate of publication of follow-up reports within seven years of the initial publication was 27%. Follow-up studies of controlled clinical studies (40%) were published more frequently than were those of laboratory or nonclinical studies (31%) or case series (22%), but these differences were not significant (p >.10). There was no statistically significant difference in follow-up publication rates based on publication bias. CONCLUSION: Only 27% of studies published as preliminary or pilot reports were subsequently followed by a more definitive publication. While the words preliminary and pilot suggest that publication of further, refined work is pending, this is often not the case. PMID- 11401812 TI - Effects of a signature on rates of change: a randomized controlled trial involving continuing education and the commitment-to-change model. AB - PURPOSE: Physicians frequently are asked to sign commitments to change practice, based upon their involvement in continuing medical education (CME) activities. Although use of the commitment-to-change model is increasingly widespread in CME, the effect of signing such commitments on rates of change is not well understood. METHOD: Immediately after a CME session, 110 physicians were asked to specify a change they intended to make in practice and to designate a level of commitment to change. To determine the effects of a signature on rates of change, physicians were randomly assigned to control (signature) and experimental (non-signature) groups. Follow-up surveys were conducted at two and three months to determine rates of change. RESULTS: In all, 88 physicians completed the first questionnaire, and 64 of them completed the follow-up. Consistent with prior studies involving the commitment-to-change model, those expressing an intention to change were significantly more likely to change on follow-up (p =.035). There was no significant difference between signature and non-signature groups (p =.99), regardless of age or gender. CONCLUSIONS: Signatures appear unimportant to assuring compliance with commitments to change used in CME conferences. A physician's behavior can be expected to change if the specified change is consistent with the physician's beliefs and sense of what is important. The relative influences of components of the commitment-to-change model require further study to determine more clearly their roles in causation and measurement. PMID- 11401813 TI - Retention and use of patient-centered interviewing skills after intensive training. AB - PURPOSE: Studies show that residents trained in patient-centered interviewing (PCI) are more effective in handling patients' emotions and are more skillful in gathering patients' data. This study evaluated the long-term use of PCI skills. METHOD: Fourteen residents received PCI training during internship, and their skills were evaluated before, immediately after, and two years after their training through directly observed patient interviews. A confidential survey evaluated the residents' actual use of PCI two years after the intensive training. Control groups of 14 interns prior to PCI training and 14 residents from another program not trained in PCI were also surveyed. RESULTS: Residents' use of PCI skills (optimization of setting, establishment of narrative thread, open-to-closed-ended questioning cone, avoid asking more than one question at a time, and (facilitation) were significantly improved, even two years after their training in PCI. The residents who received intensive block training reported using PCI techniques more frequently than did those in the control groups. However, the only significant difference in use of PCI skills between the intervention and control groups was found in reflection of patient's emotions. CONCLUSION: Medical residents retained PCI skills for two years. Further studies are needed to determine whether successful postgraduate training of physicians in PCI translates into a change in behaviors during their professional lives. PMID- 11401814 TI - Letters of recommendation: comparing perceptions of premedical advisors and admission committee members. AB - A comparison of admission committee members' and premedical advisors' perceptions of recommendation letters revealed similar ratings of the usefulness of letters and impressions of applicants. PMID- 11401815 TI - Implementation and evaluation of a telemedicine course for physician assistants. AB - Clinical telemedicine uses interactive video technologies and telecommunications networks to deliver medical consultations to distant patients and their primary care providers. Telemedicine provides real-time access to specialists whose services might not otherwise be available in rural or medically underserved areas. While recently there has been dramatic growth in the use of telemedicine, there is little evidence that telemedicine as a patient care delivery system has been incorporated into the medical school curriculum. The authors describe the current status of telemedicine in medical curricula and report on efforts at the University of Iowa to incorporate telemedicine into the curriculum of its Physician Assistant Program. PMID- 11401816 TI - Twenty years of experience using trigger films as a teaching tool. AB - Trigger films or trigger videos that depict patient-physician encounters can be used to provoke reflection, stimulate discussion, help learners confront their feelings and give learners practice in responding to challenges. For more than 20 years at the B. Rappaport Faculty of Medicine, the authors have used trigger films to teach/demonstrate the doctor-patient relationship, medical ethics, diagnostic thinking, professional behavior, and the application of the principles of the Israeli Patient Bill of Rights, and have found them to be an excellent tool for provoking active participation in small-group discussions. The authors describe how they have effectively produced and used trigger films in an Introduction to Clinical Medicine course. They highly recommend the "homemade" production of trigger films. PMID- 11401817 TI - Challenges and strategies for early professional experience: Case Western Reserve University's Family Clinic Program. AB - Through Case Western Reserve University's Family Clinic Program, students are introduced early in their education to the professional responsibilities, attributes, and interpersonal skills needed by a physician. While this is the longest running instructional program in the school's curriculum, until 1998 it had not been assessed for points of improvement. The authors used focus groups and qualitative surveys of students to identify program deficiencies and to create an educational intervention to alleviate these problems. Although the program has always been popular with students, those surveyed noted three key areas where improvements were needed: feedback from preceptors, clarification of the goals of the program, and certain aspects of dealing with patients in the clinic setting. PMID- 11401818 TI - "Physician, heal thyself": how teaching holistic medicine differs from teaching CAM. AB - The term complementary and alternative medicine (CAM) has been adopted to describe a system of health care not generally recognized as part of mainstream medical practice. It is often conflated with an older term, holistic medicine, which can briefly be defined as the art and science of healing the whole person body, mind, and spirit-in relation to that person's community and environment. Coursework in CAM is now offered in at least two thirds of U.S. medical schools. There is also a growing number of courses in the medical humanities and in spirituality and health. However, courses explicitly designed to introduce students to the principles and practices of holistic medicine are unusual. The author describes the fundamental differences between CAM and holistic medicine, highlighting holistic medicine's emphasis on the promotion of healthy lifestyles for practitioners and patients alike. He argues that offering physicians-to-be more coursework in holistic medicine could lay the groundwork for future physicians' adopting and modeling healthy lifestyles. PMID- 11401819 TI - Results of the National Resident Matching Program for 2001. PMID- 11401820 TI - Inorganic pyrophosphate generation and disposition in pathophysiology. AB - Inorganic pyrophosphate (PP(i)) regulates certain intracellular functions and extracellular crystal deposition. PP(i) is produced, degraded, and transported by specialized mechanisms. Moreover, dysregulated cellular PP(i) production, degradation, and transport all have been associated with disease, and PP(i) appears to directly mediate specific disease manifestations. In addition, natural and synthetic analogs of PP(i) are in use or currently under evaluation as prophylactic agents or therapies for disease. This review summarizes recent developments in the understanding of how PP(i) is made and disposed of by cells and assesses the body of evidence for potentially significant physiological functions of intracellular PP(i) in higher organisms. Major topics addressed are recent lines of molecular evidence that directly link decreased and increased extracellular PP(i) levels with diseases in which connective tissue matrix calcification is disordered. To illustrate in depth the effects of disordered PP(i) metabolism, this review weighs the roles in matrix calcification of the transmembrane protein ANK, which regulates intracellular to extracellular movement of PP(i), and the PP(i)-generating phosphodiesterase nucleotide pyrophosphatase family isoenzyme plasma cell membrane glycoprotein-1 (PC-1). PMID- 11401821 TI - Focus on "contrasting effects of cPLA2 on epithelial Na+ transport". PMID- 11401822 TI - A potential second permeability barrier of the epithelial Na+ channel. Focus on "point mutations in the post-M2 region of human alpha-ENaC regulate cation selectivity". PMID- 11401823 TI - ANG II-mediated inhibition of neuronal delayed rectifier K+ current: role of protein kinase C-alpha. AB - It was previously determined that ANG II and phorbol esters inhibit Kv current in neurons cultured from newborn rat hypothalamus and brain stem in a protein kinase C (PKC)- and Ca2+-dependent manner. Here, we have further defined this signaling pathway by investigating the roles of "physiological" activators of PKC and different PKC isozymes. The cell-permeable PKC activators, diacylglycerol (DAG) analogs 1,2-dioctanoyl-sn-glycerol (1 micromol/l, n = 7) and 1-oleoyl-2-acetyl-sn glycerol (1 micromol/l, n = 6), mimicked the effect of ANG II and inhibited Kv current. These effects were abolished by the PKC inhibitor chelerythrine (1 micromol/l, n = 5) or by chelation of internal Ca2+ (n = 8). PKC antisense (AS) oligodeoxynucleotides (2 micromol/l) against Ca2+-dependent PKC isoforms were applied to the neurons to manipulate the endogenous levels of PKC. PKC-alpha-AS (n = 4) treatment abolished the inhibitory effects of ANG II and 1-oleoyl-2 acetyl-sn-glycerol on Kv current, whereas PKC-beta-AS (n = 4) and PKC-gamma-AS (n = 4) did not. These results suggest that the angiotensin type 1 receptor-mediated effects of ANG II on neuronal Kv current involve activation of PKC-alpha. PMID- 11401824 TI - Hypoxic vasodilation in porcine coronary artery is preferentially inhibited by organ culture. AB - Hypoxia (95% N2-5% CO2) elicits an endothelium-independent relaxation (45-80%) in freshly dissected porcine coronary arteries. Paired artery rings cultured at 37 degrees C in sterile DMEM (pH approximately 7.4) for 24 h contracted normally to KCl or 1 microM U-46619. However, relaxation in response to hypoxia was sharply attenuated compared with control (fresh arteries or those stored at 4 degrees C for 24 h). Hypoxic vasorelaxation in organ cultured vessels was reduced at both high and low stimulation, indicating that both Ca2+-independent and Ca2+ dependent components are altered. In contrast, relaxation to G-kinase (sodium nitroprusside) or A-kinase (forskolin and isoproterenol) activation was not significantly affected by organ culture. Additionally, there was no difference in relaxation after washout of the stimulus, indicating that the inhibition is specific to acute hypoxia-induced relaxation. Simultaneous force and intracellular calcium concentration ([Ca2+]i) measurements indicate the reduction in [Ca2+]i concomitant with hypoxia at low stimulus levels in these tissue is abolished by culture. Our results indicate that organ culture at 37 degrees C specifically attenuates hypoxic relaxation in vascular smooth muscle by altering dynamics of [Ca2+]i handling and decreasing a Ca2+-independent component of relaxation. Thus organ culture can be a novel tool for investigating the mechanisms of hypoxia-induced vasodilation. PMID- 11401825 TI - Expression and characterization of the anion transporter homologue YNL275w in Saccharomyces cerevisiae. AB - A search of the yeast Saccharomyces cerevisiae genome has revealed an open reading frame, YNL275w, which encodes a 576-amino acid protein that shows sequence similarity to the family of mammalian Cl-/HCO3- anion exchangers and Na+/HCO3- cotransporters. This yeast protein also has a very similar hydropathy profile to the mammalian HCO3- transporters, indicating a similar membrane topology and structure. A V5 epitope and His6-tagged version of Ynl275wp was expressed in yeast and was localized to the plasma membrane by subcellular fractionation and immunofluorescence labeling. The protein was purified by nickel affinity chromatography and was found not to be N-glycosylated. The protein's mobility on SDS-PAGE gels was not altered by treatment with N-glycanase F, alpha mannosidase, or by mutation of each of the five consensus N-glycosylation sites. The protein did not bind to concanavalin A by lectin blotting or lectin affinity chromatography. The expressed protein bound specifically to a stilbene disulfonate inhibitor resin (SITS-Affi-Gel), and this binding could be competed by certain anions (HCO3-, Cl-, NO3-, and I-) but not by others (SO4(2-) and PO4(3 )). These results suggest that the yeast gene YNL275w encodes a nonglycosylated anion transport protein, localized to the plasma membrane. PMID- 11401826 TI - ClC-2 Cl- channels in human lung epithelia: activation by arachidonic acid, amidation, and acid-activated omeprazole. AB - ClC-2 Cl- channels represent a potential target for therapy in cystic fibrosis. Key questions regarding the feasibility of using ClC-2 as a therapeutic target are addressed in the present studies, including whether the channels are present in human lung epithelia and whether activators of the channel can be identified. Two new mechanisms of activation of human recombinant ClC-2 Cl- channels expressed in HEK-293 cells were identified: amidation with glycine methyl ester catalyzed by 1-ethyl-3(3-dimethylaminopropyl) carbodiimide (EDC) and treatment with acid-activated omeprazole. ClC-2 mRNA was detected by RT-PCR. Channel function was assessed by measuring Cl- currents by patch clamp in the presence of a cAMP-dependent protein kinase (PKA) inhibitor, myristoylated protein kinase inhibitor, to prevent PKA-activated Cl- currents. Calu-3, A549, and BEAS-2B cell lines derived from different human lung epithelia contained ClC-2 mRNA, and Cl- currents were increased by amidation, acid-activated omeprazole, and arachidonic acid. Similar results were obtained with buccal cells from healthy individuals and cystic fibrosis patients. The ClC-2 Cl- channel is thus a potential target for therapy in cystic fibrosis. PMID- 11401827 TI - Hypertonicity-induced expression of aquaporin 3 in MDCK cells. AB - Aquaporins (AQPs) are water channel proteins that participate in water transport. In the principal cells of the kidney collecting duct, water reabsorption is mediated by the combined action of AQP2 in the apical membrane and both AQP3 and AQP4 in the basolateral membrane, and the expression of AQP2 and AQP3 is regulated by antidiuretic hormone and water restriction. The effect of hypertonicity on AQP3 expression in Madin-Darby canine kidney (MDCK) epithelial cells was investigated by exposing the cells to hypertonic medium containing raffinose or NaCl. Northern blot and immunoblot analyses revealed that the amounts of AQP3 mRNA and AQP3 protein, respectively, were markedly increased by exposure of cells to hypertonicity. These effects were maximal at 12 and 24 h, respectively. Immunofluorescence and immunoelectron microscopy also demonstrated that the abundance of AQP3 protein was increased in cells incubated in hypertonic medium and that the protein was localized at the basolateral plasma membrane. These results indicate that the expression of AQP3 is upregulated by hypertonicity. PMID- 11401828 TI - Point mutations in the post-M2 region of human alpha-ENaC regulate cation selectivity. AB - We tested the hypothesis that an arginine-rich region immediately following the second transmembrane domain may constitute part of the inner mouth of the epithelial Na+ channel (ENaC) pore and, hence, influence conduction and/or selectivity properties of the channel by expressing double point mutants in Xenopus oocytes. Double point mutations of arginines in this post-M2 region of the human alpha-ENaC (alpha-hENaC) led to a decrease and increase in the macroscopic conductance of alphaR586E,R587Ebetagamma- and alphaR589E,R591Ebetagamma-hENaC, respectively, but had no effect on the single channel conductance of either double point mutant. However, the apparent equilibrium dissociation constant for Na+ was decreased for both alphaR586E,R587Ebetagamma- and alphaR589E,R591Ebetagamma-hENaC, and the maximum amiloride-sensitive Na+ current was decreased for alphaR586E,R587Ebetagamma-hENaC and increased for alphaR589E,R591Ebetagamma-hENaC. The relative permeabilities of Li+ and K+ vs. Na+ were increased 11.25- to 27.57-fold for alphaR586E,R587Ebetagamma-hENaC compared with wild type. The relative ion permeability of these double mutants and wild-type ENaC was inversely related to the crystal diameter of the permeant ions. Thus the region of positive charge is important for the ion permeation properties of the channel and may form part of the pore itself. PMID- 11401829 TI - Intercellular communication in cultured human vascular smooth muscle cells. AB - Intercellular communication through gap junction channels plays a fundamental role in regulating vascular myocyte tone. We investigated gap junction channel expression and activity in myocytes from the physiologically distinct vasculature of the human internal mammary artery (IMA, conduit vessel) and saphenous vein (SV, capacitance vessel). Northern and Western blots documented the presence of connexin43 (Cx43) in frozen tissues and cultured cells from both vessels. Northern blots also confirmed the presence of Cx40 mRNA in cultured IMA and SV myocytes. Dual whole cell patch-clamp experiments revealed that macroscopic junctional conductance was voltage dependent and characteristic of that observed for Cx43. In the majority of records, in both vessels, single-channel activity was dominated by a main-state conductance of 120 pS, with subconducting events comprising less than 10% of the amplitude histograms. However, some records showed "atypical" unitary events that had a conductance similar to Cx40 (approximately 140-160 pS), but gating behavior like that of Cx43. As such, it is conceivable that the presence and coexpression of Cx40 and Cx43 in IMA and SV myocytes may result in heteromeric channel formation. Nonetheless, in terms of gating, Cx43-like behavior clearly dominates. PMID- 11401830 TI - Regulation of I(Cl,swell) in neuroblastoma cells by G protein signaling pathways. AB - Guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) activated the I(Cl,swell) anion channel in N1E115 neuroblastoma cells in a swelling-independent manner. GTPgammaS induced current was unaffected by ATP removal and broadly selective tyrosine kinase inhibitors, demonstrating that phosphorylation events do not regulate G protein-dependent channel activation. Pertussis toxin had no effect on GTPgammaS induced current. However, cholera toxin inhibited the current approximately 70%. Exposure of cells to 8-bromoadenosine 3',5'-cyclic monophosphate did not mimic the effect of cholera toxin, and its inhibitory action was not prevented by treatment of cells with an inhibitor of adenylyl cyclase. These results demonstrate that GTPgammaS does not act through Galpha(i/o) GTPases and that Galpha(s)/Gbetagamma G proteins inhibit the channel and/or channel regulatory mechanisms through cAMP-independent mechanisms. Swelling-induced activation of I(Cl,swell) was stimulated two- to threefold by GTPgammaS and inhibited by 10 mM guanosine 5'-O-(2-thiodiphosphate). The Rho GTPase inhibitor Clostridium difficile toxin B inhibited both GTPgammaS- and swelling-induced activation of I(Cl,swell). Taken together, these findings indicate that Rho GTPase signaling pathways regulate the I(Cl,swell) channel via phosphorylation-independent mechanisms. PMID- 11401831 TI - Phosphatidylinositol 3-kinase-dependent stabilization of alpha1(I) collagen mRNA in human lung fibroblasts. AB - We investigated the role of phosphatidylinositol 3-kinase (PI3K) in the expression of alpha1(I) collagen mRNA. We report that the basal level of alpha1(I) collagen mRNA was reduced when PI3K activity was inhibited by either LY 294002 or wortmannin. These PI3K inhibitors also blocked increases of alpha1(I) collagen mRNA levels after the addition of transforming growth factor-beta. The effect of PI3K inhibition was abolished by the removal of the inhibitor or by the addition of cycloheximide. Inhibition of PI3K activity decreased the stability of the alpha1(I) collagen mRNA with no change in the rate of transcription of the alpha1(I) collagen gene as assessed by Northern blotting with actinomycin D treated fibroblasts and nuclear run-on assays. Expression of a truncated alpha1(I) collagen minigene driven by a cytomegalovirus promoter in murine fibroblasts was decreased by LY-294002 treatment. These data indicate that PI3K activation results in increased stabilization of alpha1(I) collagen mRNA. In vivo, the PI3K activity in fibroblasts may regulate basal levels of alpha1(I) collagen mRNA expression. PMID- 11401833 TI - Deficiency in parvalbumin increases fatigue resistance in fast-twitch muscle and upregulates mitochondria. AB - The soluble Ca2+-binding protein parvalbumin (PV) is expressed at high levels in fast-twitch muscles of mice. Deficiency of PV in knockout mice (PV -/-) slows down the speed of twitch relaxation, while maximum force generated during tetanic contraction is unaltered. We observed that PV-deficient fast-twitch muscles were significantly more resistant to fatigue than were the wild type. Thus components involved in Ca2+ homeostasis during the contraction-relaxation cycle were analyzed. No upregulation of another cytosolic Ca2+-binding protein was found. Mitochondria are thought to play a physiological role during muscle relaxation and were thus analyzed. The fractional volume of mitochondria in the fast-twitch muscle extensor digitorum longus (EDL) was almost doubled in PV -/- mice, and this was reflected in an increase of cytochrome c oxidase. A faster removal of intracellular Ca2+ concentration ([Ca2+]i) 200-700 ms after fast-twitch muscle stimulation observed in PV -/- muscles supports the role for mitochondria in late [Ca2+]i removal. The present results also show a significant increase of the density of capillaries in EDL muscles of PV -/- mice. Thus alterations in the dynamics of Ca2+ transients detected in fast-twitch muscles of PV -/- mice might be linked to the increase in mitochondria volume and capillary density, which contribute to the greater fatigue resistance of these muscles. PMID- 11401832 TI - [Ca2+]i as a potential downregulator of alpha2beta1-integrin-mediated A2058 tumor cell migration to type IV collagen. AB - We have investigated cellular Ca2+ regulation during A2058 human melanoma cell chemotaxis to type IV collagen (CIV). We have identified alpha2beta1-integrin as the primary mediator of A2058 cell response to CIV in vitro. Integrin ligation initiated a characteristic intracellular Ca2+ concentration ([Ca2+]i) response consisting of an internal release and a receptor-mediated Ca2+ entry. Thapsigargin (TG) pretreatment drained overlapping and CIV-inducible internal Ca2+ stores while initiating a store-operated Ca2+ release (SOCR). CIV-mediated Ca2+ entry was additive to TG-SOCR, suggesting an independent signaling mechanism. Similarly, ionophore application in a basal medium containing Ca2+ initiated a sustained influx. Elevated [Ca2+]i from TG-SOCR or ionophore significantly attenuated cell migration to CIV by recruiting the Ca2+/calcineurin mediated signaling pathway. Furthermore, low [Ca2+]i induced by EGTA application in the presence of ionophore fully restored cell motility to CIV. Together, these results suggest that [Ca2+]i signaling accompanying A2058 cell response to alpha2beta1-integrin ligation is neither necessary nor sufficient and that elevated [Ca2+]i downregulates cell motility via a calcineurin-mediated mechanism in A2058 cell chemotaxis to CIV. PMID- 11401834 TI - p21-activated kinase 1 participates in tracheal smooth muscle cell migration by signaling to p38 Mapk. AB - Cell migration contributes to many physiological processes and requires dynamic changes in the cytoskeleton. These migration-dependent cytoskeletal changes are partly mediated by p21-activated protein kinases (PAKs). At least four closely related isoforms, PAK1, PAK2, PAK3, and PAK4, exist in mammalian cells. In smooth muscle cells, little is known about the expression, activation, or ability of PAKs to regulate migration. Our study revealed the existence of three PAK isoforms in cultured tracheal smooth muscle cells (TSMCs). Additionally, we constructed adenoviral vectors encoding wild type and a catalytically inactive PAK1 mutant to investigate PAK activation and its role in TSMC migration. Stimulation of TSMCs with platelet-derived growth factor (PDGF) increased the activity of PAK1 over time. Overexpression of mutant PAK1 blocked PDGF-induced chemotactic cell migration. Phosphorylation of p38 mitogen-activated protein kinase (MAPK) in cells overexpressing wild-type PAK1 was similar to vector controls; however, p38 MAPK phosphorylation was severely reduced by overexpression of the PAK1 mutant. Collectively, these results suggest a role for PAK1 in chemotactic TSMC migration that involves catalytic activity and may require signaling to p38 MAPK among other pathways. PMID- 11401835 TI - Modulation of Na+/H+ exchange activity by Cl-. AB - Na+/H+ exchanger (NHE) activity is exquisitely dependent on the intra- and extracellular concentrations of Na+ and H+. In addition, Cl- ions have been suggested to modulate NHE activity, but little is known about the underlying mechanism, and the Cl- sensitivity of the individual isoforms has not been established. To explore their Cl- sensitivity, types 1, 2, and 3 Na+/H+ exchangers (NHE1, NHE2, and NHE3) were heterologously expressed in antiport deficient cells. Bilateral replacement of Cl- with nitrate or thiocyanate inhibited the activity of all isoforms. Cl- depletion did not affect cell volume or the cellular ATP content, which could have indirectly altered NHE activity. The number of plasmalemmal exchangers was unaffected by Cl- removal, implying that inhibition was due to a decrease in the intrinsic activity of individual exchangers. Analysis of truncated mutants of NHE1 revealed that the anion sensitivity resides, at least in part, in the COOH-terminal domain of the exchanger. Moreover, readdition of Cl- into the extracellular medium failed to restore normal transport, suggesting that intracellular Cl- is critical for activity. Thus interaction of intracellular Cl- with the COOH terminus of NHE1 or with an associated protein is essential for optimal activity. PMID- 11401836 TI - Preconditioning improves function and recovery of single muscle fibers during severe hypoxia and reoxygenation. AB - Reperfusion following prolonged ischemia induces cellular damage in whole skeletal muscle models. Ischemic preconditioning attenuates the deleterious effects. We tested whether individual skeletal muscle fibers would be similarly affected by severe hypoxia and reoxygenation (H/R) in the absence of extracellular factors and whether cellular damage could be alleviated by hypoxic preconditioning. Force and free cytosolic Ca2+ ([Ca2+]c) were monitored in Xenopus single muscle fibers (n = 24) contracting tetanically at 0.2 Hz during 5 min of severe hypoxia and 5 min of reoxygenation. Twelve cells were preconditioned by a shorter bout of H/R 1 h before the experimental trial. In preconditioned cells, force relative to initial maximal values (P/P(o)) and relative peak [Ca2+]c fell (P < 0.05) during 5 min of hypoxia and recovered during reoxygenation. In contrast, P/P(o) and relative peak [Ca2+]c fell more during hypoxia (P < 0.05) and recovered less during reoxygenation (P < 0.05) in control cells. The ratio of force to [Ca2+]c was significantly higher in the preconditioned cells during severe hypoxia, suggesting that changes in [Ca2+]c were not solely responsible for the loss in force. We conclude that 1) isolated skeletal muscle fibers contracting in the absence of extracellular factors are susceptible to H/R injury associated with changes in Ca2+ handling; and 2) hypoxic preconditioning improves contractility, Ca2+ handling, and cell recovery during subsequent hypoxic insult. PMID- 11401837 TI - Contrasting effects of cPLA2 on epithelial Na+ transport. AB - Activity of the epithelial Na+ channel (ENaC) is the limiting step for discretionary Na+ reabsorption in the cortical collecting duct. Xenopus laevis kidney A6 cells were used to investigate the effects of cytosolic phospholipase A2 (cPLA2) activity on Na+ transport. Application of aristolochic acid, a cPLA2 inhibitor, to the apical membrane of monolayers produced a decrease in apical [3H]arachidonic acid (AA) release and led to an approximate twofold increase in transepithelial Na+ current. Increased current was abolished by the nonmetabolized AA analog 5,8,11,14-eicosatetraynoic acid (ETYA), suggesting that AA, rather than one of its metabolic products, affected current. In single channel studies, ETYA produced a decrease in ENaC open probability. This suggests that cPLA2 is tonically active in A6 cells and that the end effect of liberated AA at the apical membrane is to reduce Na+ transport via actions on ENaC. In contrast, aristolochic acid applied basolaterally inhibited current, and the effect was not reversed by ETYA. Basolateral application of the cyclooxygenase inhibitor ibuprofen also inhibited current. Both effects were reversed by prostaglandin E2 (PGE2). This suggests that cPLA2 activity and free AA, which is metabolized to PGE2, are necessary to support transport. This study supports the fine-tuning of Na+ transport and reabsorption through the regulation of free AA and AA metabolism. PMID- 11401838 TI - Bcl-2 decreases voltage-gated K+ channel activity and enhances survival in vascular smooth muscle cells. AB - Cell shrinkage is an incipient hallmark of apoptosis in a variety of cell types. The apoptotic volume decrease has been demonstrated to attribute, in part, to K+ efflux; blockade of plasmalemmal K+ channels inhibits the apoptotic volume decrease and attenuates apoptosis. Using combined approaches of gene transfection, single-cell PCR, patch clamp, and fluorescence microscopy, we examined whether overexpression of Bcl-2, an anti-apoptotic oncoprotein, inhibits apoptosis in pulmonary artery smooth muscle cells (PASMC) by diminishing the activity of voltage-gated K+ (Kv) channels. A human bcl-2 gene was infected into primary cultured rat PASMC using an adenoviral vector. Overexpression of Bcl-2 significantly decreased the amplitude and current density of Kv currents (I(Kv)). In contrast, the apoptosis inducer staurosporine (ST) enhanced I(Kv). In bcl-2 infected cells, however, the ST-induced increase in I(Kv) was completely abolished, and the ST-induced apoptosis was significantly inhibited compared with cells infected with an empty adenovirus (-bcl-2). Blockade of Kv channels in control cells (-bcl-2) by 4-aminopyridine also inhibited the ST-induced increase in I(Kv) and apoptosis. Furthermore, overexpression of Bcl-2 accelerated the inactivation of I(Kv) and downregulated the mRNA expression of the pore-forming Kv channel alpha-subunits (Kv1.1, Kv1.5, and Kv2.1). These results suggest that inhibition of Kv channel activity may serve as an additional mechanism involved in the Bcl-2-mediated anti-apoptotic effect on vascular smooth muscle cells. PMID- 11401839 TI - Dynamics of nuclear localization sites for COX-2 in vascular endothelial cells. AB - We investigated the relationships among expression, activity, and spatial organization of cyclooxygenase (COX-1 and COX-2) in endothelial cells from porcine and human cerebral microvessels and from human umbilical vein. In quiescent cells, COX-1 was detected in the perinuclear zone and the cytoplasm, while COX-2 was mainly a nuclear resident possibly connected with the nuclear matrix. COX-2 immunogold labeling was situated in the nuclear envelope, at the nuclear pores, and in connection with the perichromatin regions of the nucleus, considered to be the sites of active transcription. In human endothelial cells transcriptionally activated by interleukin (IL)-1beta, the nucleus remained a major COX-2 localization site during the first 12 h of stimulation, when COX-2 expression was maximally induced. The continuous rise in prostanoid synthesis at 17-23 h of stimulation was associated with COX-2 relocation from the nucleus to the nuclear envelope and the cytoplasm. IL-1beta did not affect COX-1 expression, activity, and localization. COX-2 nuclear localization sites and trafficking between the nucleus and the cytoplasm in endothelial cells may indicate a novel function of COX-2 in regulating gene expression. PMID- 11401840 TI - Regulation of translation factors during hindlimb unloading and denervation of skeletal muscle in rats. AB - In the rat, denervation and hindlimb unloading are two commonly employed models used to study skeletal muscle atrophy. In these models, muscle atrophy is generally produced by a decrease in protein synthesis and an increase in protein degradation. The decrease in protein synthesis has been suggested to occur by an inhibition at the level of protein translation. To better characterize the regulation of protein translation, we investigated the changes that occur in various translation initiation and elongation factors. We demonstrated that both hindlimb unloading and denervation produce alterations in the phosphorylation and/or total amount of the 70-kDa ribosomal S6 kinase, eukaryotic initiation factor 2 alpha-subunit, and eukaryotic elongation factor 2. Our findings indicate that the regulation of these protein translation factors differs between the models of atrophy studied and between the muscles evaluated (e.g., soleus vs. extensor digitorum longus). PMID- 11401841 TI - COX-2 expression and cell cycle progression in human fibroblasts. AB - Cyclooxygenase-2 (COX-2) is continuously expressed in most cancerous cells where it appears to modulate cellular proliferation and apoptosis. However, little is known about the contribution of transient COX-2 induction to cell cycle progression or programmed cell death in primary cells. In this study we determined whether COX-2 regulates proliferation or apoptosis in human fibroblasts. COX-2 mRNA, protein, and prostaglandin E2 (PGE2) were not detected in quiescent cells but were expressed during the G0/G1 phase of the cell cycle induced by serum. Inhibition of COX-2 did not alter G0/G1 to S phase transition or induce apoptosis at concentrations that diminished PGE2. Addition of interleukin-1beta to serum enhanced COX-2 expression and PGE2 synthesis over that by serum alone but had no effect on the progression of these cells into S phase. Furthermore, platelet-derived growth factor drove the G0 fibroblasts into the cell cycle without inducing detectable levels of COX-2 or PGE2. Collectively, these data show that transient COX-2 expression in primary human fibroblasts does not influence cell cycle progression. PMID- 11401842 TI - Incorporation of caged cysteine and caged tyrosine into a transmembrane segment of the nicotinic ACh receptor. AB - The nonsense codon suppression technique was used to incorporate o-nitrobenzyl cysteine or o-nitrobenzyl tyrosine (caged Cys or Tyr) into the 9' position of the M2 transmembrane segment of the gamma-subunit of the muscle nicotinic ACh receptor expressed in Xenopus oocytes. The caged amino acids replaced an endogenous Leu residue that has been implicated in channel gating. ACh-induced current increased substantially after ultraviolet (UV) irradiation to remove the caging group. This represents the first successful incorporation of caged Cys into a protein in vivo and the first incorporation of caged amino acids within a transmembrane segment of a membrane protein. The bulky nitrobenzyl group does not prevent the synthesis, assembly, or trafficking of the ACh receptor. When side chains were decaged using 1-ms UV light flashes, the channels with caged Cys or caged Tyr responded with strikingly different kinetics. The increase in current upon photolysis of caged Cys was too rapid for resolution by the voltage-clamp circuit [time constant (tau) <10 ms], whereas the increase in current upon photolysis of caged Tyr was dominated by a phase with tau approximately 500 ms. Apparently, the presence of a bulky o-nitrobenzyl Tyr residue distorts the receptor into an abnormal conformation. Upon release of the caging group, the receptor relaxes, with tau approximately 500 ms, into a conformation that allows the channel to open. Tyr at the 9' position of the gamma-subunit greatly increases the ability of ACh to block the channel by binding within the channel pore. This is manifested in two ways. 1) A "rebound," or increase in current, occurs upon removal of ACh from the bathing medium; and 2) at ACh concentrations >400 microM, inward currents are decreased through the mutated channel. The ability to incorporate caged amino acids into proteins should have widespread utility. PMID- 11401843 TI - Effects of a domain peptide of the ryanodine receptor on Ca2+ release in skinned skeletal muscle fibers. AB - Mutations in the central domain of the skeletal muscle ryanodine receptor (RyR) cause malignant hyperthermia (MH). A synthetic peptide (DP4) in this domain (Leu 2442-Pro-2477) produces enhanced ryanodine binding and sensitized Ca2+ release in isolated sarcoplasmic reticulum, similar to the properties in MH, possibly because the peptide disrupts the normal interdomain interactions that stabilize the closed state of the RyR (Yamamoto T, El-Hayek R, and Ikemoto N. J Biol Chem 275: 11618-11625, 2000). Here, DP4 was applied to mechanically skinned fibers of rat muscle that had the normal excitation-contraction coupling mechanism still functional to determine whether muscle fiber responsiveness was enhanced. DP4 (100 microM) substantially potentiated the Ca2+ release and force response to caffeine (8 mM) and to low [Mg2+] (0.2 mM) in every fiber examined, with no significant effect on the properties of the contractile apparatus. DP4 also potentiated the response to submaximal depolarization of the transverse tubular system by ionic substitution. Importantly, DP4 did not significantly alter the size of the twitch response elicited by action potential stimulation. These results support the proposal that DP4 causes an MH-like aberration in RyR function and are consistent with the voltage sensor triggering Ca2+ release by destabilizing the closed state of the RyRs. PMID- 11401844 TI - Transmembrane domain length determines intracellular membrane compartment localization of syntaxins 3, 4, and 5. AB - Insulin recruits glucose transporter 4 (GLUT-4) vesicles from intracellular stores to the plasma membrane in muscle and adipose tissue by specific interactions between the vesicle membrane-soluble N-ethylmaleimide-sensitive factor attachment protein target receptor (SNARE) protein VAMP-2 and the target membrane SNARE protein syntaxin 4. Although GLUT-4 vesicle trafficking has been intensely studied, few have focused on the mechanism by which the SNAREs themselves localize to specific membrane compartments. We therefore set out to identify the molecular determinants for localizing several syntaxin isoforms, including syntaxins 3, 4, and 5, to their respective intracellular compartments (plasma membrane for syntaxins 3 and 4; cis-Golgi for syntaxin 5). Analysis of a series of deletion and chimeric syntaxin constructs revealed that the 17-amino acid transmembrane domain of syntaxin 5 was sufficient to direct the cis-Golgi localization of several heterologous reporter constructs. In contrast, the longer 25-amino acid transmembrane domain of syntaxin 3 was sufficient to localize reporter constructs to the plasma membrane. Furthermore, truncation of the syntaxin 3 transmembrane domain to 17 amino acids resulted in a complete conversion to cis-Golgi compartmentalization that was indistinguishable from syntaxin 5. These data support a model wherein short transmembrane domains (< or =17 amino acids) direct the cis-Golgi localization of syntaxins, whereas long transmembrane domains (> or =23 amino acids) direct plasma membrane localization. PMID- 11401845 TI - Moderation of skeletal muscle reperfusion injury by a sLe(x)-glycosylated complement inhibitory protein. AB - The role of the sialyl Lewis(x) (sLe(x))-decorated version of soluble complement receptor type 1 (sCR1) in moderating skeletal muscle reperfusion injury, by antagonizing neutrophil endothelial selectin interaction and complement activation, is examined. Mice underwent 2 h of hindlimb ischemia and 3 h of reperfusion. Permeability index (PI) was assessed by extravasation of 125I labeled albumin. Neutrophil depletion and complement inhibition with sCR1 reduced permeability by 72% (PI 0.81 +/- 0.10) compared with a 42% decrease (PI 1.53 +/- 0.08) observed in neutropenic mice, indicating that part of the complement mediated injury is neutrophil independent. sCR1sLe(x) treatment reduced PI by 70% (PI 0.86 +/- 0.06), an additional 20% decrease compared with sCR1 treatment (PI 1.32 +/- 0.08). Treatment with sCR1sLe(x) 0.5 and 1 h after reperfusion reduced permeability by 63% (PI 0.09 +/- 0.07) and 52% (PI 1.24 +/- 0.09), respectively, compared with the respective decreases of 41% (PI 1.41 +/- 0.10) and 32% (PI 1.61 +/- 0.07) after sCR1 treatment. Muscle immunohistochemistry stained for sCR1 only on the vascular endothelium of sCR1sLe(x)-treated mice. In conclusion, sCR1sLe(x) is more effective than sCR1 in moderating skeletal muscle reperfusion injury. PMID- 11401846 TI - Regions in the carboxy terminus of alpha-bENaC involved in gating and functional effects of actin. AB - Gating differences occur between the alpha-subunits of the bovine and rat clones of an amiloride-sensitive epithelial Na+ channel (ENaC). Deletion of the carboxy terminus of bovine alpha-ENaC (alpha-bENaC) at R567 converted the gating properties to that of rat alpha-ENaC (alpha-rENaC). The equivalent truncation in alpha-rENaC was without effect on the gating of the rat homologue. The addition of actin to ENaC channels composed of either alpha-rENaC or alpha-bENaC alone produced a twofold reduction in conductance and an increase in open probability. Neither alpha-rENaC (R613X) nor alpha-bENaC (R567X) was responsive to actin. Using a chimera consisting of alpha-rENaC1-615 and alpha-bENaC570-650, we examined several different carboxy-terminal truncation mutants plus and minus actin. When incorporated into planar bilayers, the gating pattern of this construct was identical to wild-type (wt) alpha-bENaC. Premature stop mutations proximal to E685X produced channels with gating patterns like alpha-rENaC. Actin had no effect on the E631X truncation, whereas more distal truncations all interacted with actin, as did wt alpha-bENaC. Key findings were confirmed using channels expressed in Xenopus oocytes and studied by cell-attached patch-clamp recording. Our results suggest that the site of actin regulation at the carboxy terminus of the chimera is located between residues 631 and 644. PMID- 11401847 TI - TNF receptor I is required for induction of macrophage heat shock protein 70. AB - Expression of heat shock proteins (HSP) is an adaptive response to cellular stress. Stress induces tumor necrosis factor (TNF)-alpha production. In turn, TNF alpha induces HSP70 expression. However, osmotic stress or ultraviolet radiation activates TNF-alpha receptor I (TNFR-I) in the absence of TNF-alpha. We postulated that TNF-alpha receptors are involved in the induction of HSP70 by cellular stress. Peritoneal Mphi were isolated from wild-type (WT), TNF-alpha knockout (KO), and TNFR (I or II) KO mice. Cells were cultured overnight and then heat stressed at 43 +/- 0.5 degrees C for 30 min followed by a 4-h recovery at 37 degrees C. Cellular HSP70 expression was induced by heat stress or exposure to endotoxin [lipopolysaccharide (LPS)] as determined by immunoblotting. HSP70 expression induced by either heat or LPS was markedly decreased in TNFR-I KO Mphi, whereas TNFR-II KO Mphi exhibited HSP70 expression comparable to that in WT mice. Expression of HSP70 after heat stress in TNF-alpha KO Mphi was also similar to that in WT mice, suggesting that induction of HSP70 by TNFR-I occurs independently of TNF-alpha. In addition, levels of steady-state HSP70 mRNA were similar by RT-PCR in WT and TNFR-I KO Mphi despite differences in protein expression. Furthermore, the effect of TNFR-I appears to be cell specific, since HSP70 expression in splenocytes isolated from TNFR-I KO was similar to that in WT splenocytes. These studies demonstrate that TNFR-I is required for the synthesis of HSP70 in stressed Mphi by a TNF-independent mechanism and support an intracellular role for TNFR-I. PMID- 11401848 TI - Inhibition of VRAC by c-Src tyrosine kinase targeted to caveolae is mediated by the Src homology domains. AB - We used the whole cell patch-clamp technique in calf pulmonary endothelial (CPAE) cells to investigate the effect of wild-type and mutant c-Src tyrosine kinase on I(Cl,swell), the swelling-induced Cl- current through volume-regulated anion channels (VRAC). Transient transfection of wild-type c-Src in CPAE cells did not significantly affect I(Cl,swell). However, transfection of c-Src with a Ser3Cys mutation that introduces a dual acylation signal and targets c-Src to lipid rafts and caveolae strongly repressed hypotonicity-induced I(Cl,swell) in CPAE cells. Kinase activity was dispensable for the inhibition of I(Cl,swell), since kinase deficient c-Src Ser3Cys either with an inactivating point mutation in the kinase domain or with the entire kinase domain deleted still suppressed VRAC activity. Again, the Ser3Cys mutation was required to obtain maximal inhibition by the kinase-deleted c-Src. In contrast, the inhibitory effect was completely lost when the Src homology domains 2 and 3 were deleted in c-Src. We therefore conclude that c-Src-mediated inhibition of VRAC requires compartmentalization of c-Src to caveolae and that the Src homology domains 2 and/or 3 are necessary and sufficient for inhibition. PMID- 11401849 TI - Regulation by GDI of RhoA/Rho-kinase-induced Ca2+ sensitization of smooth muscle myosin II. AB - We characterized the role of guanine nucleotide dissociation inhibitor (GDI) in RhoA/Rho-kinase-mediated Ca2+ sensitization of smooth muscle. Endogenous contents (approximately 2-4 microM) of RhoA and RhoGDI were near stoichiometric, whereas a supraphysiological GDI concentration was required to relax Ca2+ sensitization of force by GTP and guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS). GDI also inhibited Ca2+ sensitization by GTP. G14V RhoA, by alpha-adrenergic and muscarinic agonists, and extracted RhoA from membranes. GTPgammaS translocated Rho-kinase to a Triton X-114-extractable membrane fraction. GTP. G14V RhoA complexed with GDI also induced Ca2+ sensitization, probably through in vivo dissociation of GTP. RhoA from the complex, because it was reversed by addition of excess GDI. GDI did not inhibit Ca2+ sensitization by phorbol ester. Constitutively active Cdc42 and Rac1 inhibited Ca2+ sensitization by GTP. G14V RhoA. We conclude that 1) the most likely in vivo function of GDI is to prevent perpetual "recycling" of GDP. RhoA to GTP. RhoA; 2) nucleotide exchange (GTP for GDP) on complexed GDP. RhoA/GDI can precede translocation of RhoA to the membrane; 3) activation of Rho-kinase exposes a hydrophobic domain; and 4) Cdc42 and Rac1 can inhibit Ca2+ sensitization by activated GTP. RhoA. PMID- 11401850 TI - Modeling of transcellular Ca transport in rat duodenum points to coexistence of two mechanisms of apical entry. AB - Employing realistic parameters, we have demonstrated that a relatively simple mathematical model can reproduce key features of steady-state Ca2+ transport with the assumption of two mechanisms of Ca2+ entry: a channel-like flux and a carrier mediated transport. At low luminal [Ca2+] (1-5 mM), facilitated entry dominates and saturates with Km = 0.4 mM. At luminal [Ca2+] of tens of millimolar, apical permeability is dominated by the channel flux that in turn is regulated by cytosolic Ca2+. The model reproduces the linear relationship between maximum Ca2+ transport rate and intestinal calbindin D9K (CaBP) content. At luminal [Ca2+] > 50 mM, local sensitivity analysis shows transcellular transport to be most sensitive to variations in CaBP. At low luminal [Ca2+], transport becomes sensitive to apical entry regulation. The simulations have been run within the Virtual Cell modeling environment, yielding the time course of external Ca2+ and spatiotemporal distributions of both intracellular Ca2+ and CaBP. Coexistence of two apical entry mechanisms accords with the properties of the duodenal Ca2+ transport protein CaT1 and the epithelial Ca2+ channel ECaC. PMID- 11401851 TI - Apical P2Y4 purinergic receptor controls K+ secretion by vestibular dark cell epithelium. AB - It was previously shown that K+ secretion by vestibular dark cell epithelium is under control of G protein-coupled receptors of the P2Y family in the apical membrane that are activated by both purine and uridine nucleotides (P2Y2, P2Y4, or P2Y6). The present study was conducted to determine the subtype of purinergic receptor and to test whether these receptors undergo desensitization. The transepithelial short-circuit current represents electrogenic K+ secretion and was found to be reduced by UTP, ATP, and diadenosine tetraphosphate, but not UDP. Neither pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS, 30 microM) nor suramin (100 microM) inhibited the effect of UTP. The potencies of the agonists were consistent with rodent P2Y4 and P2Y2, but not P2Y6, receptors. The ineffectiveness of suramin was consistent with P2Y4, but not P2Y2. Transcripts for both P2Y2 and P2Y4 were found in vestibular labyrinth. Sustained exposure to ATP or UTP for 15 min caused a constant depression of short-circuit current with no apparent desensitization. The results support the conclusion that regulation of K+ secretion across vestibular dark cell epithelium occurs by P2Y4 receptors without desensitization of the response. PMID- 11401852 TI - KChAP/Kvbeta1.2 interactions and their effects on cardiac Kv channel expression. AB - KChAP and voltage-dependent K+ (Kv) beta-subunits are two different types of cytoplasmic proteins that interact with Kv channels. KChAP acts as a chaperone for Kv2.1 and Kv4.3 channels. It also binds to Kv1.x channels but, with the exception of Kv1.3, does not increase Kv1.x currents. Kvbeta-subunits are assembled with Kv1.x channels; they exhibit "chaperone-like" behavior and change gating properties. In addition, KChAP and Kvbeta-subunits interact with each other. Here we examine the consequences of this interaction on Kv currents in Xenopus oocytes injected with different combinations of cRNAs, including Kvbeta1.2, KChAP, and either Kv1.4, Kv1.5, Kv2.1, or Kv4.3. We found that KChAP attenuated the depression of Kv1.5 currents produced by Kvbeta1.2, and Kvbeta1.2 eliminated the increase of Kv2.1 and Kv4.3 currents produced by KChAP. Both KChAP and Kvbeta1.2 are expressed in cardiomyocytes, where Kv1.5 and Kv2.1 produce sustained outward currents and Kv4.3 and Kv1.4 generate transient outward currents. Because they interact, either KChAP or Kvbeta1.2 may alter both sustained and transient cardiac Kv currents. The interaction of these two different classes of modulatory proteins may constitute a novel mechanism for regulating cardiac K+ currents. PMID- 11401853 TI - Smooth muscle length-dependent PI(4,5)P2 synthesis and paxillin tyrosine phosphorylation. AB - We studied effects of increasing the length of porcine trachealis muscle on 5.5 microM carbachol (CCh)-evoked phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] synthesis and other parameters of phosphatidylinositol (PI) turnover. PI(4,5)P2 resynthesis rates in muscle held at 1.0 optimal length (L(o)), measured over the first 6 min of CCh stimulation, were 140 +/- 12 and 227 +/- 14% of values found in muscle held at 0.5 L(o) and in free-floating muscle, respectively. Time dependent changes in cellular masses of PI(4,5)P2, PI, and phosphatidic acid, and PI resynthesis rates, were also altered by the muscle length at which contraction occurred. In free-floating muscle, CCh did not evoke increases in tyrosine phosphorylated paxillin (PTyr-paxillin), an index of beta1-integrin signaling; however, there were progressive increases in PTyr-paxillin in muscle held at 0.5 and 1.0 L(o) during contraction, which correlated with increases in PI(4,5)P2 synthesis rates. These data indicate that PI(4,5)P2 synthesis rates and other parameters of CCh-stimulated inositol phospholipid turnover are muscle length dependent and provide evidence that supports the hypothesis that length-dependent beta1-integrin signals may exert control on CCh-activated PI(4,5)P2 synthesis. PMID- 11401854 TI - Smad4 mediates activation of mitogen-activated protein kinases by TGF-beta in pancreatic acinar cells. AB - Transforming growth factor-beta (TGF-beta) inhibits pancreatic acinar cell growth. In many cell types, TGF-beta mediates its growth inhibitory effects by activation of Smad proteins. Recently, it has been reported that Smad proteins may interact with the mitogen-activated protein (MAP) kinase signaling pathways. In this study, we report on the interactions between the TGF-beta and MAP kinase signaling pathways in isolated rat pancreatic acinar cells. TGF-beta activated the MAP kinases extracellular signal-related kinases (ERKs) and p38 in pancreatic acinar cells, but had no effect on c-jun NH2-terminal kinase activity. Activation of MAP kinase by TGF-beta was maximal 4 h after treatment. The ability of TGF beta to activate ERKs was concentration dependent and dependent on protein synthesis. TGF-beta's stimulation of ERK activation was blocked by PD-98059, an inhibitor of MAP kinase kinase 1, and by adenoviral transfer of dominant negative RasN17. Furthermore, adenoviral-mediated expression of dominant negative Smad4 blocked the ability of TGF-beta to activate acinar cell MAP kinase, demonstrating that this activation is downstream of Smads. The biological relevance of ERK activation by TGF-beta was indicated by demonstrating that inhibition of ERK signaling by PD-98059 blocked the ability of TGF-beta to activate the transcription factor activator protein-1. These studies provide new insight into the signaling mechanisms by which TGF-beta mediates biological actions in pancreatic acinar cells. PMID- 11401855 TI - Functional aspects of creatine kinase isoenzymes in endothelial cells. AB - To characterize the isoenzyme distribution of creatine kinase (CK) in endothelial cells (ECs) and its functional role during substrate depletion, ECs from aorta (AECs) and microvasculature (MVECs) of pig and rat were studied. In addition, high- energy phosphates were continuously monitored by (31)P NMR spectroscopy in pig AECs attached to microcarrier beads. CK activity per milligram of protein in rat AECs and MVECs (0.08 +/- 0.01 and 0.15 +/- 0.08 U/mg, respectively) was <3% of that of cardiomyocytes (6.46 +/- 1.02 U/mg). Rat and pig AECs and MVECs displayed cytosolic BB-CK, but no MM-CK. Gel electrophoresis of mitochondrial fractions of rat and pig ECs indicated the presence of mitochondrial Mi-CK, mostly in dimeric form. The presence of Mi(a)-CK was demonstrated by indirect immunofluorescence staining using Mi(a)-CK antibodies. When perifused with creatine-supplemented medium, phosphocreatine (PCr) continuously increased with time (1.2 +/- 0.6 nmol x h(-1) x mg x protein(-1)), indicating creatine uptake and CK activity. Glucose withdrawal from the medium induced a rapid decrease in PCr, which was fully reversible on glucose addition, demonstrating temporal buffering of an energy deficit. Because both cytosolic and mitochondrial CK isoforms are present in ECs, the CK system may also contribute to energy transduction ("shuttle hypothesis"). PMID- 11401856 TI - Polyamine transport system mediates agmatine transport in mammalian cells. AB - Agmatine is a biogenic amine with the capacity to regulate a number of nonreceptor-mediated functions in mammalian cells, including intracellular polyamine content and nitric oxide generation. We observed avid incorporation of agmatine into several mammalian cell lines and herein characterize agmatine transport in mammalian cells. In transformed NIH/3T3 cells, agmatine uptake is energy dependent with a saturable component indicative of carrier-mediated transport. Transport displays an apparent Michaelis-Menten constant of 2.5 microM and a maximal velocity of 280 pmol x min(-1) x mg(-1) protein and requires a membrane potential across the plasma membrane for uptake. Competition with polyamines, but not cationic molecules that utilize the y+ system transporter, suppresses agmatine uptake. Altering polyamine transporter activity results in parallel changes in polyamine and agmatine uptake. Furthermore, agmatine uptake is abrogated in a polyamine transport-deficient human carcinoma cell line. These lines of evidence demonstrate that agmatine utilizes, and is dependent on, the polyamine transporter for cellular uptake. The fact that this transport system is associated with proliferation could be of consequence to the antiproliferative effects of agmatine. PMID- 11401857 TI - Neutrophils injure cultured skeletal myotubes. AB - The purpose of the study was to test the hypothesis that neutrophils can injure cultured skeletal myotubes. Human myotubes were grown and then cultured with human blood neutrophils. Myotube injury was quantitatively and qualitatively determined using a cytotoxicity (51Cr) assay and electron microscopy, respectively. For the 51Cr assay, neutrophils, under non-in vitro-stimulated and N-formylmethionyl-leucyl-phenylalanine (FMLP)-stimulated conditions, were cultured with myotubes at effector-to-target cell (E:T) ratios of 10, 30, and 50 for 6 h. Statistical analyses revealed that myotube injury was proportional to the E:T ratio and was greater in FMLP-stimulated conditions relative to non-in vitro-stimulated conditions. Transmission electron microscopy, using lanthanum as an extracellular tracer, revealed in cocultures a diffuse appearance of lanthanum in the cytoplasm of myotubes and a localized appearance within cytoplasmic vacuoles of myotubes. These observations and their absence in control cultures (myotubes only) suggest that neutrophils caused membrane rupture and increased myotube endocytosis, respectively. Myotube membrane blebs were prevalent in scanning and transmission electron micrographs of cultures consisting of neutrophils and myotubes (E:T ratio of 5) and were absent in control cultures. These data support the hypothesis that neutrophils can injure skeletal myotubes in vitro and may indicate that neutrophils exacerbate muscle injury and/or delay muscle regeneration in vivo. PMID- 11401858 TI - Anti-adrenergic effects of nitric oxide donor SIN-1 in rat cardiac myocytes. AB - We studied how the nitric oxide (NO*) donor 3-morpholinosydnonimine (SIN-1) alters the response to beta-adrenergic stimulation in cardiac rat myocytes. We found that SIN-1 decreases the positive inotropic effect of isoproterenol (Iso) and decreases the extent of both cell shortening and Ca2+ transient. These effects of SIN-1 were associated with an increased intracellular concentration of cGMP, a decreased intracellular concentration of cAMP, and a reduction in the levels of phosphorylation of phospholamban (PLB) and troponin I (TnI). The guanylyl cyclase inhibitor 1H-8-bromo-1,2,4-oxadiazolo (3,4-d)benz(b)(1,4)oxazin 1-one (ODQ) was not able to prevent the SIN-1-induced reduction of phosphorylation levels of PLB and TnI. However, the effects of SIN-1 were abolished in the presence of superoxide dismutase (SOD) or SOD and catalase. These data suggest that, in the presence of Iso, NO-related congeners, rather than NO*, are responsible for SIN-1 effects. Our results provide new insights into the mechanism by which SIN-1 alters the positive inotropic effects of beta adrenergic stimulation. PMID- 11401859 TI - Ionomycin causes activation of p38 and p42/44 mitogen-activated protein kinases in human neutrophils. AB - Many receptor-linked agents that prime or activate the NADPH oxidase in polymorphonuclear neutrophils (PMNs) elicit changes in cytosolic Ca2+ concentration and activate mitogen-activated protein (MAP) kinases. To investigate the role of Ca2+ in the activation of p38 and p42/44 MAP kinases, we examined the effects of the Ca2+-selective ionophore ionomycin on priming and activation of the PMN oxidase. Ionomycin caused a rapid rise in cytosolic Ca2+ that was due to both a release of cytosolic Ca2+ stores and Ca2+ influx. Ionomycin also activated (2 microM) and primed (20-200 nM) the PMN oxidase. Dual phosphorylation of p38 MAP kinase and phosphorylation of its substrate activating transcription factor-2 were detected at ionomycin concentrations that prime or activate the PMN oxidase, while dual phosphorylation of p42/44 MAP kinase and phosphorylation of its substrate Elk-1 were elicited at 0.2-2 microM. SB-203580, a p38 MAP kinase antagonist, inhibited ionomycin-induced activation of the oxidase (68 +/- 8%, P < 0.05) and tyrosine phosphorylation of 105- and 72-kDa proteins; conversely, PD-98059, an inhibitor of MAP/extracellular signal-related kinase 1, had no effect. Treatment of PMNs with thapsigargin resulted in priming of the oxidase and activation of p38 MAP kinase. Chelation of cytosolic but not extracellular Ca2+ completely inhibited ionomycin activation of p38 MAP kinase, whereas chelation of extracellular Ca2+ abrogated activation of p42/44 MAP kinase. These results demonstrate the importance of changes in cytosolic Ca2+ for MAP kinase activation in PMNs. PMID- 11401860 TI - Characterization of a novel 132-bp exon of the human maxi-K channel. AB - The large-conductance Ca2+-activated voltage-dependent K+ channel (maxi-K channel) induces a significant repolarizing current that buffers cell excitability. This channel can derive its diversity by alternative splicing of its transcript-producing isoforms that differ in their sensitivity to voltage and intracellular Ca2+. We have identified a novel 132-bp exon of the maxi-K channel from human myometrial cells that encodes 44 amino acids within the first intracellular loop of the channel protein. Distribution analysis reveals that this exon is expressed predominantly in human smooth muscle tissues with the highest abundance in the uterus and aorta and resembles the previously reported distribution of the total maxi-K channel transcript. Single-channel K+ current measurements in fibroblasts transfected with the maxi-K channel containing this novel 132-bp exon demonstrate that the presence of this insert attenuates the sensitivity to voltage and intracellular Ca2+. Alternative splicing to introduce this 132-bp exon into the maxi-K channel may elicit another mode to modulate cell excitability. PMID- 11401861 TI - Are polymorphisms the answer in hypersensitivity pneumonitis? PMID- 11401862 TI - How best to measure airway responsiveness. PMID- 11401863 TI - Intravascular catheter use. How to tell when the medicine is worse than the malady. PMID- 11401864 TI - Defining the lung's response to endotoxin. PMID- 11401865 TI - Is the measurement of sleepiness the Holy Grail of sleep medicine? PMID- 11401866 TI - Adrenal insufficiency in critically ill patients. PMID- 11401867 TI - Practice expense costs in pulmonary and critical care practices. Is providing patient care still economically feasible? PMID- 11401868 TI - Major histocompatibility complex and tumor necrosis factor-alpha polymorphisms in pigeon breeder's disease. AB - Pigeon breeders disease (PBD) is caused by the exposure of a susceptible host to avian antigens. However, genetic factors determining individual predisposition are unknown. In this work, polymorphisms of the major histocompatibility complex (MHC) class II alleles and tumor necrosis factor alpha (TNF-alpha) promoter were evaluated in 44 patients with PBD, 99 healthy unrelated controls (HC), and 50 exposed but asymptomatic subjects (EAS). MHC typing was performed by PCR-specific sequence oligonucleotide analysis, and TNF-alpha polymorphism at -238 and -308 positions by amplification refractory mutation system-PCR. PBD patients showed a significant increase of the alleles HLA-DRB1*1305 (p < 0.001, OR = 15.4, 95% CI = 3.18-102.6 [HC], and OR = 17.05, 95% CI = 2.25-357.8 [EAS]) and HLA-DQB1*0501 (p < 0.05, OR = 2.93, 95% CI = 1.21-7.15 [HC], and OR = 2.96, 95% CI = 1.0-9.14 [EAS]). A decrease of HLA-DRB1*0802 was also noticed in patients when compared with both control groups (p < 0.05). Haplotype analysis revealed an increase of DRB1*1305-DQB1*0301 and a decrease of DRB1*0802-DQB1*0402. PBD patients had an increased frequency of TNF-2(-)(308) compared with both control groups (p < 0.05). Patients exhibiting the TNF-2(-)(308) allele were younger (33.9 +/- 14.6 versus 44.2 +/- 10.4 yr; p < 0.05), and displayed more lymphocytes in their bronchoalveolar lavages (88.0 +/- 12.1 versus 68.9 +/- 17.2; p < 0.05). These results suggest that genetic factors located within the MHC region contribute to the development of PBD. PMID- 11401869 TI - Role of molds in farmer's lung disease in Eastern France. AB - Farmer's lung disease (FLD) is common in the east of France. In the absence of the primary recognized FLD agent, Saccharopolyspora rectivirgula, its etiology remains unknown. A prospective case-control study was performed to find the etiology of FLD in this area. Eleven patients were matched with 11 healthy control farmers. Twenty-two urban subjects constituted the nonexposed control group. Microorganisms from cowshed air and fodder were identified and counted. The antigens of the microorganisms most frequently isolated at the 22 farms were used for serological tests. Farms of patients with FLD contained more Absidia corymbifera than those of healthy farmers (p < 0.05 in air, p < 0.01 in fodder). Electrosyneresis, performed with A. corymbifera somatic antigen, differentiated 9 of 11 patients with FLD from control subjects (p < 0.01). Other significant results were obtained with Eurotium amstelodami (p < 0.01) and Wallemia sebi (p < 0.05). In contrast, no significant results were obtained with the other seven antigens tested, including S. rectivirgula. Absidia corymbifera and, to a lesser degree, W. sebi or E. amstelodami are likely to be the main causes of FLD in this area. Modifications in working conditions over time could explain the emergence of these new contributing etiologies. PMID- 11401870 TI - Highly activated T-cell receptor AV2S3(+) CD4(+) lung T-cell expansions in pulmonary sarcoidosis. AB - Sarcoidosis is a systemic disorder of unknown origin, primarily affecting the lungs. The granulomatous inflammation is driven by the interplay between various molecules and cells, including T cells. Previously, our group reported a close correlation between lung-restricted T-cell receptor (TCR) AV2S3 CD4-positive T cell expansions and HLA-DR17 in active sarcoidosis. The aim of this study was to characterize phenotypically such AV2S3 lung T cells, to obtain more information about the state of activation of this intriguing T-cell subset. Bronchoalveolar lavage (BAL) was performed on sarcoidosis patients with active disease and on healthy control subjects (HC). The expression of activation and subset markers was evaluated and compared between BAL AV2S3-positive and AV2S3-negative T cells of patients with lung-restricted AV2S3 T-cell expansions, and between BAL and peripheral blood lymphocytes (PBL) of patients and HC. The frequency of cells expressing activation markers CD26, CD28, CD69, and HLA-DR was enhanced in AV2S3 positive versus AV2S3-negative BAL CD4(+) T-cell subsets. In contrast, CD25 (Il 2R) and CD27 were expressed at lower levels by the AV2S3-positive CD4(+) lung T cells. Our data confirm a substantial activation of BAL CD4(+) T cells of patients with sarcoidosis. Furthermore, the AV2S3 CD4-positive lung cells display a pattern of activation markers, suggesting that they are significantly more activated compared with lung CD4(+) T cells expressing other TCR V gene segments as well as compared with BAL CD4(+) T cells of HC. These results support our hypothesis of an ongoing and selective stimulation of AV2S3 T cells by a specific antigen and the participation of this subset in the inflammatory process in the lungs of patients with sarcoidosis. PMID- 11401871 TI - PC(20) adenosine 5'-monophosphate is more closely associated with airway inflammation in asthma than PC(20) methacholine. AB - Inhalation of a direct stimulus such as histamine or methacholine is generally used to measure bronchial hyperresponsiveness (BHR). Provocation with adenosine 5'-monophosphate (AMP), an indirect airway challenge, has been suggested to be a better marker of airway inflammation than direct challenges. However, so far little information on this subject is available. The aim of our study was to assess whether the concentration of AMP causing the FEV(1) to drop by 20% (PC(20)) is more closely associated with inflammatory parameters in asthma than PC(20) methacholine. In 120 patients with atopic asthma (median FEV(1) 81% predicted [pred], median age 27 yr), PC(20) methacholine and PC(20) AMP as well as sputum induction, blood sampling, and measurement of nitric oxide in exhaled air were performed. PC(20) methacholine was predominantly predicted by FEV(1) %pred (explained variance [ev] = 18%) with the percentage of peripheral blood monocytes being a weak additional independent predictor (total ev = 23%). By contrast, PC(20) AMP was predominantly predicted by the percentage of eosinophils in sputum (ev = 25%), while FEV(1) %pred was only an additional independent predictor (total ev = 36%). PC(20) AMP reflects more closely the extent of airway inflammation due to asthma than PC(20) methacholine. PMID- 11401872 TI - Distal lung dysfunction at night in nocturnal asthma. AB - We have previously shown that patients with nocturnal worsening of asthma (nocturnal asthma) exhibit increased parenchymal inflammation at night. To evaluate the functional significance of this parenchymal inflammation, 10 subjects with nocturnal asthma (NA), four subjects with non-nocturnal asthma (NNA), and four normal control subjects underwent bronchoscopy with measurement of peripheral airways resistance (Rp) at 4:00 P.M. and at 4:00 A.M. Employing a wedged bronchoscope technique, Rp was measured. Flow was stopped, and the pressure reached after 10 s of decay was termed the plateau pressure. The time constant of this decay (tau) was measured, and the peripheral compliance (Cp) was calculated as tau/Rp. The NA group exhibited the highest Rp values at 4:00 P.M. and at 4:00 A.M. as compared with the NNA and control groups, but all groups were significantly different from each other at 4:00 P.M.: NA, 0.113 +/- 0.02 cm H(2)O/ml/min; NNA, 0.033 +/- 0.005 cm H(2)O/ml/min; Control subjects, 0.010 +/- 0.001 cm H(2)O/ ml/min; p = 0.0001; and at 4:00 A.M.: NA, 0.129 +/- 0.023 cm H(2)O/ ml/min; NNA, 0.035 +/- 0.007 cm H(2)O/ml/min; Control subjects, 0.009 +/- 0.002 cm H(2)O/ml/min; p = 0.0003. None of the groups exhibited statistically significant differences in Rp between 4:00 P.M. and 4:00 A.M. The plateau pressure increased significantly from 4:00 P.M. to 4:00 A.M., but only in the NA group (7.7 +/- 0.9 cm H(2)O at 4:00 P.M. versus 16.9 +/- 4.6 cm H(2)O at 4:00 A.M.; p = 0.0004). Cp was decreased in the NA group as compared with the NNA and control groups at both 4:00 P.M. (p = 0.0003) and 4:00 A.M. (p = 0.003). The Rp positively correlated with the residual volume at both 4:00 P.M. (r = 0.71, p = 0.004) and 4:00 A.M. (r = 0.59, p = 0.03). We conclude that the distal lung units, specifically the collateral channels, and may be functionally altered at night in NA. PMID- 11401873 TI - Extended exhaled NO measurement differentiates between alveolar and bronchial inflammation. AB - Lower respiratory tract inflammation can be detected by measuring exhaled nitric oxide (NO) concentration at a single exhalation flow rate, but this does not differentiate between alveolar and bronchial NO production. We assessed alveolar NO concentration and bronchial NO flux with an extended method of measuring exhaled NO at several exhalation flow rates in 40 patients with asthma, 17 patients with alveolitis, and 57 healthy control subjects. Bronchial NO flux was higher in asthma (2.5 +/- 0.3 nl/s, p < 0.001) than in alveolitis (0.7 +/- 0.1 nl/s) and healthy control subjects (0.7 +/- 0.1 nl/s). Alveolar NO concentration was higher in alveolitis (4.1 +/- 0.3 ppb, p < 0.001) than in asthma (1.1 +/- 0.2 ppb) and healthy control subjects (1.1 +/- 0.1 ppb). In asthma, bronchial NO flux correlated with serum level of eosinophil protein X (EPX) (r = 0.60, p < 0.001) and bronchial hyperresponsiveness (r = 0.55, p < 0.001). In alveolitis, alveolar NO concentration correlated inversely with pulmonary diffusing capacity (r = 0.55, p = 0.022) and pulmonary restriction. Glucocorticoid treatment or allergen avoidance normalized bronchial NO flux in asthma and decreased alveolar NO concentration toward normal in alveolitis. In conclusion, extended exhaled NO measurement can be used to separately assess alveolar and bronchial inflammation and to assess disease activity/severity in asthma and alveolitis. PMID- 11401874 TI - Lung function 5 yr after lung volume reduction surgery for emphysema. AB - Current datum more than 2 yr after lung volume reduction surgery (LVRS) for emphysema is limited. This prospective study evaluates pre-LVRS baseline and 5-yr results in 26 symptomatic patients (mean age 67 +/- 6 yr) (mean +/- SD) who underwent bilateral, targeted upper lobe stapled LVRS using video-assisted thoracoscopy. Baseline forced expiratory volume in 1 s (FEV(1)) was 0.7 +/- 0.2 L (mean +/- SD), 29 +/- 10% predicted. Following LVRS, with none lost to follow-up, mortality due to respiratory failure at 0.5, 1, 2, 3, 4, and 5 yr was 4%, 4%, 19%, 31%, 46%, and 58%, respectively. Increase above baseline for FEV(1) > 200 ml and/or FVC > 400 ml at 1, 2, 3, 4, and 5 yr post-LVRS was noted in 73%, 46%, 35%, 27%, and 8% of all patients; decrease in dyspnea grade >/= 1 in 88%, 69%, 46%, 27%, and 15%; and elimination of initial oxygen dependence in 18 patients in 78%, 50%, 33%, 22%, and 0%, respectively. Expiratory airflow improved due to the increase in both lung elastic recoil and small airway intraluminal caliber. Five patients decreased FEV(1) 141 +/- 60 ml/yr and FVC 102 +/- 189 ml/yr over 3.8 +/- 1.2 yr post-LVRS, similar to their pre-LVRS rate of decline. In the 11 patients who survived 5 yr, at 0.5-1.0 yr post-LVRS peak increase in FEV(1) was 438 +/- 366 ml, with a decline of 149 +/- 157 ml the following year and 78 +/- 59 ml/yr over 4.0-4.5 yr. Bilateral LVRS provided palliative clinical and physiological improvement in 9 of 26 patients at 3 yr, 7 at 4 yr, and 2 at 5 yr. PMID- 11401875 TI - Multivariable assessment of the 6-min walking test in patients with chronic obstructive pulmonary disease. AB - Functional exercise tolerance in patients with chronic obstructive pulmonary disease (COPD) is often assessed by the 6-min walking test (6MWT). To assess if the use of multiple factors adds to walking distance in describing performance in the 6MWT, an exploratory factor analysis was performed on physiological measurements and dyspnea ratings recorded during testing. Eighty-three patients with mild to severe COPD performed repeated 6MWTs before inpatient pulmonary rehabilitation. Factor analysis on 15 variables yielded a stable four-factor structure explaining 78.4% of the total variance. Recorded heart rate variables contributed to factor 1 (heart rate pattern), walking distance, heart rate increase, and decrease contributed to factor 2 (endurance capacity), oxygen desaturation variables contributed to factor 3 (impairment of oxygen transport), and dyspnea and effort variables contributed to factor 4 (perceived symptoms). Walking distance decreased in half of the 53 patients measured posttreatment, but self-perceived change in exercise tolerance improved in 84% and was explained by change in walking distance, by less desaturation, and by less dyspnea (R(2) = 0.55, p = 0.005). Qualitative analysis showed that 29 of 53 patients improved in three or four factors. Performance in the 6MWT can be described with four statistically independent and clinically interpretable factors. Because clinically relevant changes consist of more than only walking distance, assessment of functional exercise tolerance in patients with COPD improves by reporting multiple variables. PMID- 11401876 TI - Occupation, chronic bronchitis, and lung function in young adults. An international study. AB - We studied the relationship between occupational exposures, chronic bronchitis, and lung function in a general population survey in 14 industrialized countries, including 13,253 men and women aged 20 to 44 yr. We studied associations between occupational group, occupational exposures, bronchitis symptoms (cough and phlegm production for at least 3 mo each year), FEV(1), and nonspecific bronchial responsiveness (NSBR) separately in lifetime nonsmokers, cigarette smokers, and ex-smokers. Occupational exposure to vapors, gas, dust, or fumes, estimated with a job exposure matrix (JEM), was associated with chronic bronchitis among current smokers only (prevalence ratio (PR): 1.2 to 1.7). The interaction of occupational exposure with smoking, however, was not statistically significant (p > 0.1). Self reported exposure was related to chronic bronchitis in all smoking groups. An increased risk for chronic bronchitis was found in agricultural, textile, paper, wood, chemical, and food processing workers, being more pronounced in smokers. Lung function and NSBR were not clearly related to occupational exposures. Findings were similar for asthmatic and nonasthmatic subjects. In conclusion, occupational exposures contributed to the occurrence of chronic (industrial) bronchitis in young adults. Fixed airflow limitation was not evident, probably due to the relatively young age of this population. PMID- 11401877 TI - Defective natural killer and phagocytic activities in chronic obstructive pulmonary disease are restored by glycophosphopeptical (inmunoferon). AB - We have investigated both modifications in natural (innate) immunity caused by chronic obstructive pulmonary disease (COPD) and the effects of a glycophosphopeptical immunomodulator (Inmunoferon) treatment on COPD-associated immunoalterations. In a double-blinded clinical trial, 60 patients with COPD received glycophosphopeptical or placebo during 90 consecutive days at oral doses of 3 g/d. Fifty-six sex- and age-matched healthy control subjects were included as a reference group for immunologic parameters. Peripheral blood natural killer (PBNK) cell cytotoxic activity and phagocytic activity of peripheral monocytes/macrophages (Mo/Ma) and polymorphonuclear (PMN) cells were assessed at baseline and then again at the end of treatments. We found both PBNK activity and phagocytic activity to be significantly decreased in patients with COPD compared with levels in healthy volunteers. The treatment with glycophosphopeptical provoked significant stimulatory effects on PBNK cytotoxic activity. This stimulation was not mediated by an increase in CD3(-)CD56(+) NK cells. Further, glycophosphopeptical significantly increased the percentage of monocytes and PMNs that phagocytize Escherichia coli in vitro, as well as increased phagocytic indices. We conclude that peripheral blood cells of patients with COPD show clear defects in natural immunity that are partially rescued by glycophosphopeptical. PMID- 11401878 TI - Outcomes of primary and catheter-related bacteremia. A cohort and case-control study in critically ill patients. AB - We studied a cohort of 2201 patients hospitalized in 15 French intensive care units (ICUs) for > or = 48 h during a 4-mo period to assess the incidence and outcomes of primary and definite catheter-related bloodstream (CRB) or secondary nosocomial bloodstream infection (NBSI). Variables associated with ICU death and duration of stay were determined by logistic regression, and attributable mortality and length of stay (LOS) from a nested matched case-control (96 pairs) study, stratified on the source of bacteremia. Bacteremia occurred in 5% (95% CI 4.1-6%) of patients with > or = 48 h ICU stay. Primary, CRB, and secondary NBSI accounted for 29%, 26%, and 45% of the 111 episodes, respectively. NBSI was associated with a markedly increased risk of death (OR = 4.6; 95% CI 2.9-7.1) and an attributable mortality of 35% (95% CI, 28%-47%). In the case-control study, the excess mortality was 20% (p = 0.03) in patients with primary bacteremia and CRB, and 55% (p < 0.001) for secondary bacteremia; in patients with CRB only, the excess mortality was 11.5%. The median excess ICU LOS in survivors of NBSI was 9.5 d, and was similar, irrespective of its source. The risk of mortality associated with primary and catheter-related bacteremia appears much lower than that of secondary bacteremia, but is sizable, and the excess LOS incurred by the various categories of bacteremia is comparable. Differentiating catheter-related bacteremia from both primary and other secondary bacteremia appears warranted in studies conducted in critically ill patients. PMID- 11401879 TI - Local inflammatory responses following bronchial endotoxin instillation in humans. AB - To study local lung inflammation, 34 subjects had endotoxin (1-4 ng/kg) instilled into a lung segment and saline instilled into a contralateral segment followed by bronchoalveolar lavage (BAL) at 2 h, 6 h, 24 h, or 48 h. Endotoxin instillation resulted in a focal inflammatory response with a distinct time course. An early phase (2 h to 6 h) revealed an increase in neutrophils (p = 0.0001) with elevated cytokines (tumor necrosis factor [TNF]-alpha, TNF receptors [TNFR], interleukin [IL]-1beta, IL-1 receptor antagonist, IL-6, granulocyte-colony-stimulating factor [G-CSF], all p < or = 0.002, but no change in IL-10) and chemokines (IL-8, epithelial neutrophil activating protein-78, monocyte chemotactic protein-1, macrophage inflammatory protein [MIP]-1alpha, MIP-1beta, all p < or = 0.001, but no change in growth-regulated peptide-alpha). A later phase (24 h to 48 h) showed increased neutrophils, macrophages, monocytes, and lymphocytes (all p < or = 0.02), and a return to basal levels of most mediators. Elevated levels of inflammatory markers (TNFR(1), TNFR(2), L-selectin, lactoferrin, and myeloperoxidase) persisted in the BAL at 48 h (p < or = 0.001). Increased permeability to albumin occurred throughout both phases (p = 0.001). Blood C reactive protein, serum amyloid A, IL-6, IL-1ra, G-CSF, but not TNF-alpha increased by 8 h (all p < or = 0.008). The local pulmonary inflammatory response to endotoxin has a unique qualitative and temporal profile of inflammation compared with previous reports of intravenous endotoxin challenges. This model provides a means to investigate factors that initiate, amplify, and resolve local lung inflammation. PMID- 11401880 TI - Septic shock and respiratory failure in community-acquired pneumonia have different TNF polymorphism associations. AB - Genetic factors are likely to contribute to the variable presentation of community-acquired pneumonia (CAP). The purpose of this prospective cohort study was to determine whether the LTalpha+250 (TNFbeta+250) and TNFalpha-308 gene polymorphisms are associated with different presentations of CAP. Septic shock (SS) was defined using American College of Chest Physicians/Society of Critical Care Medicine (ACCP-SCCM) criteria. Type I respiratory failure (T1RF) was defined as an O(2) saturation on room air of < 90% with a normal PCO(2). A total of 280 patients were genotyped; 31 had SS, 80 had T1RF. Genotype proportions are given in the order of AA/GA/ GG. The proportion of patients in each genotype developing SS was as follows: LTalpha+250 0.19/0.07/0.09 (p = 0.01 AA versus non-AA); TNFalpha-308 0.16/0.06/0.12 (p = NS). Carrying at least one AA (tumor necrosis factor [TNF] high secretor) genotype had an 18.0% risk of SS versus 6.8% (p = 0.006). GG homozygotes (TNF low secretors) at both loci had only a 2.9% risk of SS. Septic shock was associated with the LTalpha+250:TNFalpha-308 A:G haplotype but not the A:A haplotype, suggesting that LTalpha+250 is a marker, rather than a causative polymorphism. Carriage of the G:G haplotype had a significant protective effect against the development of septic shock (p = 0.011). T1RF was not associated with LTalpha+250 AA genotype. In the absence of septic shock, there was a significant trend to greater T1RF in patients with LTalpha+250 GG (TNFalpha hyposecretor) genotype (p = 0.03). Our finding of different genotype associations for SS and T1RF has important implications for immunotherapy in both CAP and sepsis, as well as for the definition of the systemic inflammatory response syndrome (SIRS). PMID- 11401881 TI - Comparative effects of red blood cell transfusion and increasing blood flow on tissue oxygenation in oxygen supply-dependent conditions. AB - Red blood cell (RBC) transfusion is usually administered to improve oxygen delivery (DO(2)) in order to sustain tissue oxygen demand. However, this practice is not supported by firm clinical or experimental data. Using a randomized two period crossover design, this study compared the efficacy of "fresh" RBC transfusion and increased blood flow to restore tissue oxygenation in oxygen supply-dependent conditions. In 12 ketamine-anesthetized mongrel dogs submitted to nonpulsatile normothermic cardiopulmonary bypass, DO(2) was reduced by a progressive decrease in pump flow. DO(2) dependency was defined as an O(2) uptake (V O(2)) decrease by more than 15% from baseline value. Then, intervention consisted of a 40% increase in DO(2) obtained either by transfusion of "fresh" dog's RBC (stored < 3 d) or by increase in pump flow. Animals received both interventions sequentially in a random order, while O(2) saturation was maintained constant. In O(2) supply-dependent conditions, rising pump flow from 1.6 +/- 0.4 to 2.7 +/- 0.7 L/ min increased DO(2) from 5.4 +/- 1.1 to 9.0 +/- 1.3 ml/kg/min (p < 0.01) and V O(2) from 3.5 +/- 0.4 to 4.1 +/- 0.5 ml/kg/min (p = 0.02). "Fresh" RBC transfusion, which increased the hemoglobin concentration from 6.4 +/- 0.9 to 11.1 +/- 1.3 g/dl, increased DO(2) from 5.4 +/- 1.2 to 9.0 +/- 1.4 ml/kg/min (p < 0.01) and V O(2) from 3.6 +/- 0.4 to 4.1 +/- 0.5 ml/kg/min (p = 0.02). There was no difference in V O(2) resulting from both interventions. In oxygen supply-dependent conditions, "fresh" RBC transfusion and increased blood flow are equally effective in restoring tissue oxygenation. PMID- 11401882 TI - Influence of tidal volume on alveolar recruitment. Respective role of PEEP and a recruitment maneuver. AB - Both reduction in tidal volume (VT) and alveolar recruitment may be important to limit ventilator-associated lung injury during mechanical ventilation of patients with the acute respiratory distress syndrome (ARDS). The aim of this study was to assess the risk of alveolar derecruitment associated with VT reduction from 10 to 6 ml/kg. Whether this VT-related derecruitment could be reversed, either by a recruitment maneuver or by an increase in positive end-expiratory pressure (PEEP) level, was also investigated. Fifteen patients with ARDS were successively ventilated using conventional VT (CVT = 10 +/- 1 ml/kg) and low VT (LVT = 6 +/- 1 ml/ kg); total PEEP (PEEPtot) was individually set at the lower inflection point (Plip) of the pressure-volume curve (PEEPtot = 11 +/- 4 cm H(2)O). Pressure volume curves were recorded from zero PEEP (ZEEP) and from PEEP, and recruited volume (Vrec) was calculated as the volume difference between the two curves for a given pressure. Despite a similar PEEPtot, Vrec was significantly lower with LVT than with CVT, indicating low VT-induced alveolar derecruitment. Reduction in VT was associated with a reduced Sa(O(2)). In 10 patients, Vrec was also measured before and after a recruitment maneuver (two sustained inflations at 45 cm H(2)O), and after an increase in PEEP (by 4 cm H(2)O). Low VT-induced derecruitment was reversed by a recruitment maneuver and by increasing PEEP. We conclude that a reduction in VT could be responsible for alveolar derecruitment, which may be transiently reversed by a reexpansion maneuver or prevented by a PEEP increase above Plip. PMID- 11401883 TI - Physiologic effects of negative pressure ventilation in acute exacerbation of chronic obstructive pulmonary disease. AB - To assess the physiologic effects of continuous negative extrathoracic pressure (CNEP), negative pressure ventilation (NPV), and negative extrathoracic end expiratory pressure (NEEP) added to NPV in patients with acute exacerbation of chronic obstructive pulmonary disease (COPD), we measured in seven patients ventilatory pattern, arterial blood gases, respiratory mechanics, and pressure- time product of the diaphragm (PTPdi) under four conditions: (1) spontaneous breathing (SB); (2) CNEP (-5 cm H(2)O); (3) NPV; (4) NPV plus NEEP. CNEP and NPV were provided by a microprocessor-based iron lung capable of thermistor triggering. Compared with SB, CNEP improved slightly but significantly Pa(CO(2 ))and pH, and decreased PTPdi (388 +/- 59 versus 302 +/- 43 cm H(2)O. s, respectively, p < 0.05) and dynamic intrinsic positive end-expiratory pressure (PEEPi) (4.6 +/- 0.5 versus 2.1 +/- 0.3 cm H(2)O, respectively, p < 0.001). NPV increased minute ventilation (V E), improved arterial blood gases, and decreased PTPdi to 34% of value during SB (p < 0.001). NEEP added to NPV further slightly decreased PTPdi and improved patient-ventilator interaction by reducing dynamic PEEPi and nontriggering inspiratory efforts. We conclude that CNEP and NPV, provided by microprocessor-based iron lung, are able to improve ventilatory pattern and arterial blood gases, and to unload inspiratory muscles in patients with acute exacerbation of COPD. PMID- 11401884 TI - Microsleep during a simplified maintenance of wakefulness test. A validation study of the OSLER test. AB - Daytime somnolence is both a symptom in many patients and a prevalent complaint in the general population, but its objective assessment remains elusive. The current available tests are technically complex and thus inadequate for routine clinical use or epidemiological studies. A simplified behavioral maintenance of wakefulness test (OSLER test) has been recently described that could allow for widespread availability of objective measurements of this symptom. We verified the occurrence of (micro)sleep (episodes of sleep of > or = 3 s duration) during the performance of the OSLER test in 10 normal subjects after a non-sleep deprived night and a sleep-deprived night in randomized order. Sleep was assessed electrophysiologically according to standard methods. The OSLER test (mean of four measurements) was significantly shorter after the sleep-deprived night (25 min versus 38 min). Single missed stimuli were frequent with or without (micro)sleep, but (micro)sleep was almost always present when four or more consecutive stimuli were missed. The sensitivity and specificity of the test in detecting sleep (of > or = 3 s duration) are 85% and 94%, respectively. The total number of missed stimuli per minute duration of the test could add valuable information to the simpler mean test result. Sensitivity and specificity are not altered when only three measurements are performed. We conclude that the OSLER test appears as a simple, easy, and reliable method to objectively assess daytime somnolence. PMID- 11401885 TI - Relation of measures of sleep-disordered breathing to neuropsychological functioning. AB - Sleep-disordered breathing (SDB) has been associated with neuropsychological (NP) deficits. The extent to which such effects are attributable to unmeasured confounders or selection biases, or are manifest across a range of SDB is unclear. The relationship of SDB with a broad range of NP functions was examined in 100 volunteers with a spectrum of SDB and without underlying comorbidity. Factor analysis suggested that the NP tests could be summarized as four constructs: declarative memory, signal discrimination, working memory, and set shifting. These factors plus vigilance were dependent variables. Independent variables were age, the respiratory disturbance index (RDI), a sleepiness score, the arousal index, and sleep-associated hypoxemia. Factors "declarative memory" (measuring 25% of the common variance, alpha = 0.95), "signal discrimination" (10% variance, alpha = 0.70), and "working memory" (9% variance, alpha = 0.52) were each significantly, linearly predicted by hypoxemia and/or the RDI, with no evidence for significant threshold effects. SDB measures accounted for 4-6% of the variance in NP constructs. In contrast, sleepiness best predicted vigilance. Thus, adverse exposures (hypoxemia or RDI) during sleep may negatively influence NP functions in a dose-response relationship, and, other than vigilance, these effects may not be directly attributable to sleepiness. PMID- 11401886 TI - Impact of obstructive sleep apnea on left ventricular mass and diastolic function. AB - We wished to determine if obstructive sleep apnea (OSA) is associated with increased left ventricular mass (LVM) and impaired left ventricular diastolic function (LVDF) independently of coexisting obesity, hypertension (HTN), and diabetes mellitus (DM). Patients without primary cardiac disease, referred for evaluation of OSA (n = 533), had overnight polysomnography and Doppler echocardiography while awake. Patients were divided, according to the apnea hypopnea index (AHI), into an OSA group (AHI > or = 5/h, n = 353) and a non-OSA group (AHI < 5/h, n = 180). In men, LVM was greater in the OSA group (98.9 +/- 25.6 versus 92.3 +/- 22.5 g/m, p = 0.023) despite exclusion of those with HTN and DM. A similar trend was noted in women. Regression analysis revealed that LVM was correlated with body mass index (BMI) (beta = 0.480, p < 0.0005), age (beta = 0.16, p = 0.001), and the presence of HTN (beta = 0.137, p = 0.003) in men and with BMI (beta = 0.501, p < 0.0005) in women, but not with AHI or oxygen saturation during sleep. The ratio of peak early filling velocity to peak late filling velocity (E/A), an index of LVDF, was similar in both groups (1.28 +/- 0.32 versus 1.34 +/- 0.31, p = 0.058); it was correlated with age (beta = -0.474, p < 0.0005), but not with AHI or oxygen saturation during sleep. We conclude that OSA is not associated with increased LVM or impaired LVDF independently of obesity, HTN, or advancing age. PMID- 11401887 TI - Diaphragm activation during exercise in chronic obstructive pulmonary disease. AB - Although it has been postulated that central inhibition of respiratory drive may prevent development of diaphragm fatigue in patients with chronic obstructive pulmonary disease (COPD) during exercise, this premise has not been validated. We evaluated diaphragm electrical activation (EAdi) relative to maximum in 10 patients with moderately severe COPD at rest and during incremental exhaustive bicycle exercise. Flow was measured with a pneumotachograph and volume by integration of flow. EAdi and transdiaphragmatic pressures (Pdi) were measured using an esophageal catheter. End-expiratory lung volume (EELV) was assessed by inspiratory capacity (IC) maneuvers, and maximal voluntary EAdi was obtained during these maneuvers. Minute ventilation (V E) was 12.2 +/- 1.9 L/min (mean +/- SD) at rest, and increased progressively (p < 0.001) to 31.0 +/- 7.8 L/min at end exercise. EELV increased during exercise (p < 0.001) causing end-inspiratory lung volume to attain 97 +/- 3% of TLC at end-exercise. Pdi at rest was 9.4 +/- 3.2 cm H(2)O and increased during the first two thirds of exercise (p < 0.001) to plateau at about 13 cm H(2)O. EAdi was 24 +/- 6% of voluntary maximal at rest and increased progressively during exercise (p < 0.001) to reach 81 +/- 7% at end exercise. In conclusion, dynamic hyperinflation during exhaustive exercise in patients with COPD reduces diaphragm pressure-generating capacity, promoting high levels of diaphragm activation. PMID- 11401888 TI - Maintenance of cAMP in non-heart-beating donor lungs reduces ischemia-reperfusion injury. AB - Studies suggest that pulmonary vascular ischemia-reperfusion injury (IRI) can be attenuated by increasing intracellular cAMP concentrations. The purpose of this study was to determine the effect of IRI on capillary permeability, assessed by capillary filtration coeficient (Kfc), in lungs retrieved from non-heart-beating donors (NHBDs) and reperfused with the addition of the beta(2)-adrenergic receptor agonist isoproterenol (iso), and rolipram (roli), a phosphodiesterase (type IV) inhibitor. Using an in situ isolated perfused lung model, lungs were retrieved from NHBD rats at varying intervals after death and either ventilated with O(2) or not ventilated. The lungs were reperfused with Earle's solution with or without a combination of iso (10 microM) and roli (2 microM). Kfc, lung viability, and pulmonary hemodynamics were measured. Lung tissue levels of adenine nucleotides and cAMP were measured by HPLC. Combined iso and roli (iso/roli) reperfusion decreased Kfc significantly (p < 0.05) compared with non iso/roli-reperfused groups after 2 h of postmortem ischemia. Total adenine nucleotide (TAN) levels correlated with Kfc in non-iso/roli-reperfused (r = 0.89) and iso/roli-reperfused (r = 0.97) lungs. cAMP levels correlated with Kfc (r = 0.93) in iso/roli-reperfused lungs. Pharmacologic augmentation of tissue TAN and cAMP levels might ameliorate the increased capillary permeability observed in lungs retrieved from NHBDs. PMID- 11401889 TI - Bleomycin-induced chronic lung damage does not resemble human idiopathic pulmonary fibrosis. AB - Administration of bleomycin into the lungs of experimental animals has been utilized as a model to understand human pulmonary fibrosis. Most of the studies, however, have focused on early stages of the lung reaction. We hypothesized that chronic stages of the model may not mimic idiopathic pulmonary fibrosis, since in preliminary studies, lung volume and compliance were not decreased. Eight male Sprague-Dawley rats receiving intratracheal bleomycin (0.5 U/100 g body weight) underwent measurement of FRC, inspiratory capacity, and lung compliance 120 d later. Lung histologic changes were evaluated using light microscopy. Eight rats without intervention served as controls. Results show that our model, in early stages, has histologic changes no different from those previously described elsewhere. In chronic stages, however, the model does not behave as a restrictive syndrome: FRC is normal or increased, whereas lung compliance is normal. Focal peribronchiolar inflammation and fibrosis associated with paracicatricial emphysematous changes are the main histologic features of long-term lung remodeling after bleomycin. We conclude that while the chronic stages of the model may be informative in understanding mechanisms of fibrosis, care should be taken not to extrapolate to human idiopathic pulmonary fibrosis. We speculate that the model might resemble a particular subgroup of human interstitial lung disease, namely, those involving peribronchiolar structures. PMID- 11401890 TI - Diaphragm injury in individuals with airflow obstruction. AB - The purpose of this study was to describe the nature of diaphragm injury, to quantify the injury and number of macrophages at the light microscopic level, and to determine their association with airflow obstruction in humans. Partial thickness diaphragm biopsies were obtained from 21 subjects going for thoracotomy surgery (FEV(1): 74 +/- 34% predicted; range: 16 to 122% predicted). Cross sections cut from frozen diaphragm were processed with H&E or processed for immunohistochemistry using the monoclonal antibody Ber-MAC3 (DAKO Corp., Carpinteria, CA) to label macrophages. Area fractions (A(A)) or the proportions of the cross- sectional area were determined by point counting all viable fields of H&E-stained diaphragm cross sections. A(A) were 66.2 +/- 9.0% for normal muscle, 17.6 +/- 7.2% for abnormal muscle, and 16.3 +/- 4.2% for connective tissue. Percent predicted FEV(1) was inversely related to the A(A) of abnormal muscle (r = -0.53, p < 0.01) and directly related to the A(A) of normal muscle (r = 0.37, p < 0.05). The number of macrophages was not related to % predicted FEV(1) (mean +/- SD: 0.41 +/- 0.18/fiber; 52 +/- 19/mm(2)). We conclude that increasing severity of airflow obstruction is associated with an increased A(A) of abnormal diaphragm and a decreased A(A) of normal diaphragm. PMID- 11401891 TI - Intratracheal administration of activated protein C inhibits bleomycin-induced lung fibrosis in the mouse. AB - It is well recognized that activation of the coagulation system plays an important role in bleomycin (BLM)-induced lung injury and fibrosis. The protein C (PC) pathway is an important regulator of the coagulation system. In this study, we evaluated the bronchoalveolar lavage fluid (BALF) concentration of activated PC (APC) and the therapeutic effect of the intratracheal administration of APC on BLM-induced lung fibrosis in mice. APC levels in BALF were significantly lower in BLM-treated animals than in the saline-treated group. Fibrotic changes were progressive in mice treated with BLM and intratracheal instillation of vehicle (BLM/Veh) after 14 and 21 d of BLM infusion. Compared with the BLM/Veh group, histologic findings on Days 14 and 21 in mice treated with BLM and intratracheal instillation of APC (BLM/APC) showed less fibrotic lesions in the subpleural and central areas of the lung. The mean Aschcroft's fibrosis score in the BLM/Veh group was significantly (p < 0.05) higher than in the BLM/APC group. The lung hydroxyproline content on Day 21 was significantly higher (p < 0.05) in the BLM/Veh group (1.78 +/- 0.07 micromol/lung weight) than in the BLM/APC (1.30 +/- 0.06 micromol/lung weight) group. The ratio of plasminogen activator activity to thrombin level in BALF was significantly increased in the BLM/APC group compared with the BLM/ Veh group on Day 21. The expression of tumor necrosis factor-alpha and interleukin-1beta was significantly decreased in the lungs of the BLM/APC group compared with the BLM/Veh group on Day 14 after BLM infusion. These results showed that intratracheal APC administration inhibits the development of lung fibrosis in BLM-induced lung injury, giving further support to the important role that the PC pathway plays in the mechanism of lung fibrosis. PMID- 11401892 TI - Eotaxin expression after segmental allergen challenge in subjects with atopic asthma. AB - Expression of pulmonary eotaxin protein and mRNA was determined in six subjects with atopic asthma and five nonatopic normal subjects. Levels of eotaxin expression and eosinophil mobilization were compared before and after segmental allergen challenge in subjects with atopic asthma. In the absence of allergen challenge, we found significantly higher levels of eotaxin in the bronchoalveolar lavage (BAL) fluid of subjects with asthma than in that of normal subjects (25 +/ 3 versus 15 +/- 2 pg/ml, p < 0.05). BAL eotaxin levels increased after segmental allergen challenge in all six subjects with atopic asthma tested, with a mean increase from 22 +/- 4 to 53 +/- 10 pg/ml (p = 0.013). Segmental allergen challenge was associated with a significant increase in the percentage of BAL macrophages and eosinophils that were immunopositive for eotaxin. Eotaxin mRNA was detectable by northern analysis in BAL cells exclusively from allergen challenged segments. Allergen- induced increases in eotaxin levels were strongly associated with increases in BAL eosinophil recovery (r(2) = 0.88, p = 0.0036). Segmental allergen challenge also increased eotaxin expression in airway epithelial and endothelial cells obtained by endobronchial biopsy. These findings demonstrate, for the first time, that the airways of subjects with allergic asthma respond to allergen by increasing eotaxin expression. The tissue loci of eotaxin expression, the levels of eotaxin recovered in BAL fluid, and the association of eotaxin levels with eosinophil mobilization suggest either that eotaxin plays a mechanistic role in allergen-induced airway eosinophilia or that it serves as a biomarker for the causal mechanisms. PMID- 11401893 TI - Long-term inhalation of high-dose nitric oxide increases intraalveolar activation of coagulation system in mice. AB - Inhalation of nitric oxide (NO) is useful for the treatment of patients with pulmonary hypertension. However, the potential toxicity of inhaled NO is still unclear. Coagulation activation plays an important role in lung injury. We assessed the effect of low- and high-dose inhaled NO on the coagulation system in the intraalveolar space of mice. The animals were assigned to five groups (n = 6): [RA] group, mice exposed to fresh air alone; [RA+2 ppm NO] group, fresh air and 2 ppm NO; [RA+40 ppm NO] group, fresh air and 40 ppm NO; [RA+2 ppm NO+O(2)] group, fresh air, 2 ppm NO and O(2); and [RA+40 ppm NO+O(2)] group, fresh air, 40 ppm NO and O(2). Each group was treated for 3 wk. Lung specimens of [RA+40 ppm NO] and [RA+40 ppm NO+O(2)] groups showed significant nitrotyrosine immunoreactivity. BALF concentrations of total protein, thrombin and soluble tissue factor were significantly increased in mice of [RA+40 ppm NO] and [RA+40 ppm NO+O(2)] groups compared with [RA] group. However, BALF concentrations of total protein, thrombin, and soluble tissue factor were not significantly increased in mice of [RA+2 ppm NO] and [RA+2 ppm NO+O(2)] groups compared with [RA] group. Lung tissue factor mRNA expression was higher in the high-dose NO group than in the low-dose NO group. NO donor increased significantly tissue factor activity on alveolar epithelial cells. This study has shown for the first time that long-term inhalation of high, but not low, concentration of NO may activate the clotting system by increasing the lung expression of tissue factor. PMID- 11401894 TI - Evidence that systemic gentamicin suppresses premature stop mutations in patients with cystic fibrosis. AB - Here we report the effects of gentamicin treatment on cystic fibrosis transmembrane regulator (CFTR) production and function in CF airway cells and patients with CF with premature stop mutations. Using immunocytochemical and functional [6-methoxy-N- (3-sulfopropyl) quinolinium (SPQ)-based] techniques, ex vivo exposure of airway cells from stop mutation CF patients led to the identification of surface-localized CFTR in a dose-dependent fashion. Next, five patients with CF with stop mutations and five CF control subjects were treated with parenteral gentamicin for 1 wk, and underwent repeated in vivo measures of CFTR function (nasal potential difference [PD] measurements and sweat chloride [Cl(-)] testing). During the treatment period, the number of nasal PD readings in the direction of Cl(-) secretion was increased approximately 3-fold in the stop mutation patient group compared with controls (p < 0.001), and four of five stop mutation patients with CF had at least one reading during gentamicin treatment with a Cl(-) secretory response of more than -5 mV (hyperpolarized). A response of this magnitude was not seen in any of the CF control subjects (p < 0.05). In an independent series of experiments designed to test the ability of repeat nasal PDs to detect wild-type CFTR function, evidence of Cl(-) secretion was seen in 88% of control (non-CF) nasal PDs, and 71% were more than -5 mV hyperpolarized. Together, these results suggest that gentamicin treatment can suppress premature stop mutations in airway cells from patients with CF, and produce small increases in CFTR Cl(-) conductance (as measured by the nasal PD) in vivo. PMID- 11401895 TI - Exhaled markers of pulmonary disease. PMID- 11401896 TI - Bronchopulmonary dysplasia. PMID- 11401897 TI - Guidelines for the management of adults with community-acquired pneumonia. Diagnosis, assessment of severity, antimicrobial therapy, and prevention. PMID- 11401898 TI - The role of hypoventilation and ventilation-perfusion redistribution in oxygen induced hypercapnia during acute exacerbations of chronic obstructive pulmonary disease. PMID- 11401899 TI - Effect of unplanned extubation on outcome of mechanical ventilation. PMID- 11401900 TI - Biologic variability in mechanical ventilation in a canine oleic acid lung injury model. PMID- 11401901 TI - Complications of evaluating nodules. PMID- 11401902 TI - The problem of dose-response and therapeutic ratio of inhaled steroids. PMID- 11401903 TI - Fev(6) as a surrogate for FVC: authors should have included ROC-curve analyses. PMID- 11401905 TI - Genetic determinants of serum prostate-specific antigen levels in healthy men from a multiethnic cohort. AB - We recently reported an association between prostate cancer risk and polymorphisms in the prostate-specific antigen (PSA) and androgen receptor (AR) genes. The purpose of this study is to test whether these two polymorphisms, AR CAG and PSA ARE1, influence serum PSA levels in healthy men. Serum PSA and the two genotypes were assayed for 420 healthy men from a multiethnic cohort, and regression models were fit to estimate the effects of AR CAG genotype and PSA ARE1 genotype on serum PSA levels. Predicted serum PSA decreased 3.5% with each additional AR CAG repeat decile (P = 0.01). Serum PSA was also associated with PSA ARE1 genotype, with PSA levels higher among men with the PSA AA genotype compared with men with the AG or GG genotypes (P = 0.02). The relationship between serum PSA level and AR CAG length differed according to PSA genotype (P = 0.049): for genotype GG, the slope was not significantly different from zero (P = 0.74); for genotype AG, serum PSA increased 4.5% with each decrease of one CAG repeat decile (P = 0.03); for genotype AA, serum PSA increased 7% with each decrease of one CAG repeat decile (P = 0.02). These results indicate that in healthy men, genetic variants in the PSA and AR genes contribute to variation in serum PSA levels. Men with the PSA AA genotype and short AR CAG alleles have, on average, higher serum PSA levels. PMID- 11401906 TI - Biomarkers of polycyclic aromatic hydrocarbon-DNA damage and cigarette smoke exposures in paired maternal and newborn blood samples as a measure of differential susceptibility. AB - Human and experimental evidence indicates that the developing fetus may be more susceptible than the adult to the effects of certain carcinogens, including some polycyclic aromatic hydrocarbons (PAHs). Factors that can modulate susceptibility include proliferation rates, detoxification capabilities, and DNA repair capacity. Biomarkers can facilitate quantification of age-related susceptibility among human populations. In this study, we report on three biomarkers measured in paired blood samples collected at birth from 160 Polish mothers and newborns: 70 pairs from Krakow (a city with high air pollution including PAHs) and 90 pairs from Limanowa (an area with lower ambient pollution but greater indoor coal use). Biomarkers were: WBC aromatic-DNA adducts by (32)P-postlabeling and PAH-DNA adducts by ELISA (as indicators of DNA damage from PAHs and other aromatics) and plasma cotinine (as an internal dosimeter of cigarette smoke). Correlations were assessed by Spearman's rank test, and differences in biomarker levels were assessed by the Wilcoxon signed-ranks test. A significant correlation between paired newborn/maternal samples was seen for aromatic-DNA adduct levels (r = 0.3; P < 0.001) and plasma cotinine (r = 0.8; P < 0.001) but not PAH-DNA adduct levels (r = 0.14; P = 0.13). Among the total cohort, levels of the three biomarkers were higher in newborn samples compared with paired maternal samples. The difference was significant for aromatic-DNA adduct levels (16.6 +/- 12.5 versus 14.21 +/- 15.4/10(8) nucleotides; P = 0.002) and plasma cotinine (14.2 +/- 35.5 versus 8.3 +/- 24.5 ng/ml; P < 0.001) but not for PAH-DNA adduct levels (7.9 +/- 9.9 versus 5.9 +/- 8.2/10(8) nucleotides; P = 0.13). When analyses were restricted to the 80 mother/newborn pairs from whom the blood sample was drawn concurrently (within 1 h of each other), levels of all of the three biomarkers were significantly higher in the newborn compared with paired maternal blood samples (P < 0.05). Results suggest reduced detoxification capabilities and increased susceptibility of the fetus to DNA damage, especially in light of experimental evidence that transplacental exposures to PAHs are 10-fold lower than paired maternal exposures. The results have implications for risk assessment, which currently does not adequately account for sensitive subsets of the population. PMID- 11401907 TI - Individual differences in urinary cotinine levels in Japanese smokers: relation to genetic polymorphism of drug-metabolizing enzymes. AB - Urinary cotinine, one of the main metabolites of nicotine, has been widely used as a biomarker for assessment of direct or passive exposure to cigarette smoke. However, there is wide variation of the cotinine level among smokers who smoke the same number of cigarettes. To use urinary cotinine as a proper exposure biomarker for cigarette smoke, interindividual variations of cotinine formation must be considered. Therefore, we studied the effects of genetic polymorphisms in drug metabolic enzymes on urinary cotinine levels among 190 male Japanese smokers (ages 19-66 years; mean, 40.6 years). Genetic polymorphisms in cytochrome P-450s (CYP1A1, CYP2A6, CYP2E1), and aldehyde dehydrogenase 2 (ALDH2) were determined by analyzing DNA isolated from peripheral blood. Cotinine in morning spot urine was analyzed by high-performance liquid chromatography. Lifestyle, i.e., smoking, alcohol consumption, and intake of coffee or tea, was examined using a questionnaire. The number of cigarettes smoked and CYP2A6 polymorphism were significantly associated with the urinary cotinine level. Especially, the urinary cotinine levels was drastically lower in CYP2A6-deleted homozygous (CYP2A6*4/*4) subjects than in CYP2A6*1 allele-positive subjects. The polymorphism in the CYP2E1 5'-flanking region was related to the urinary cotinine level in intermediate smokers (who smoke 11-20 cigarettes/day; P < 0.01). Polymorphisms in CYP1A1 or ALDH2, and consumption of alcohol, coffee, or tea were not associated with the urinary cotinine level. PMID- 11401908 TI - Identification of retinamides that are more potent than N-(4 hydroxyphenyl)retinamide in inhibiting growth and inducing apoptosis of human head and neck and lung cancer cells. AB - The synthetic retinoid, N-(4-hydroxyphenyl)retinamide (4HPR), which is currently being evaluated in clinical trials for cancer prevention and therapy, inhibits the growth of a variety of malignant cells through induction of apoptosis. However, in the majority of tumor cells, this inhibitory effect of 4HPR requires high concentrations (>1 microM), which exceed the peak plasma level measured in humans. In the present study, we compared and contrasted the effects of several synthetic retinamides on the growth of human lung and head and neck cancer cells in vitro. We found that some retinamides, especially N-(2 carboxyphenyl)retinamide (2CPR), exhibited better growth inhibitory effects than 4HPR in some of the cell lines. 2CPR exerted potent growth inhibitory effects in 5 of 10 head and neck cancer cell lines and in 1 of 10 lung cancer cell lines (IC(50), <0.8 microM). 2CPR (1 microM) induced apoptosis ranging from 10 to 60% in four of five cell lines, whereas 4HPR was ineffective at the same concentration. Unlike 4HPR, 2CPR (up to 10 microM) failed to induce reactive oxygen species production in these sensitive cell lines but could activate caspases 3 and 7 as well as increase poly(ADP-ribose)polymerase cleavage. Interestingly, the effect of 2CPR on cell growth could be suppressed by the specific retinoic acid receptor pan antagonist AGN193109. Our results suggest that 2CPR acts via retinoic acid receptors and may be a good candidate for prevention and treatment of some head and neck and lung cancers. PMID- 11401909 TI - p53 protein accumulation and genomic instability in head and neck multistep tumorigenesis. AB - Head and neck cancer develops in a multistep process and is associated with increasing frequencies of p53 alterations and with increasing genomic instability. To study the relationship of p53 alterations and genomic instability during head and neck tumorigenesis, we analyzed p53 protein expression and chromosome 9 and 17 polysomy in 48 squamous cell carcinomas of the head and neck and their adjacent normal epithelium (31 sites), hyperplastic (24 sites), and dysplastic lesions (26 sites). Normal oral epithelium obtained from seven nonsmoking, cancer-free individuals served as negative controls. Six (19%) of 31 lesions in adjacent normal epithelium, 7 (29%) of 24 hyperplastic lesions, 12 (46%) of 26 dysplastic lesions, and 28 (58%) of 48 squamous cell carcinomas expressed p53. In contrast, no normal control epithelium had detectable p53 expression. To determine the relationship between dysregulated p53 expression and genomic instability during tumorigenesis, we compared p53 immunohistochemistry distributions and chromosome polysomy levels (by chromosome in situ hybridization) in different histological groups associated with tissue progression. Although the degree of chromosome polysomy increased for all of the groups during histological progression, lesions with dysregulated p53 expression showed nearly 2-4-fold increased levels of chromosome polysomy. This trend was significant for dysplastic lesions (P = 0.005 and P = 0.002 for chromosomes 9 and 17, respectively) and for squamous cell carcinoma (P = 0.005 and P = 0.002 for chromosomes 9 and 17, respectively). Image analysis studies for 28 p53-expressing tumors and their adjacent premalignant lesions demonstrated a strong spatial correlation between stepwise transitions from low to high p53 expression and increased chromosome polysomy frequencies in 13 (46%) of 28 cases. These findings suggest that altered p53 expression is associated with increased genetic instability in preneoplastic epithelium and may play a driving force for increasing the rate of accumulation of genetic events during head and neck tumorigenesis. PMID- 11401910 TI - Diabetes and endometrial cancer in the Iowa women's health study. AB - Diabetes has been associated with increased risk of endometrial cancer in some epidemiological studies. Body mass index (BMI) and other measures of obesity have been associated positively with both diabetes and endometrial cancer. It is not clear whether or not the association of diabetes with endometrial cancer is explained entirely by obesity. Thus, we sought to test the hypothesis that diabetes is not associated with endometrial cancer independent of obesity. We examined the association between self-reported diabetes (onset at >30 years of age) and incident endometrial cancer in a prospective cohort study of 24,664 postmenopausal women in Iowa. Over 12 years of follow-up, 346 cases occurred among the cohort at risk. Data were analyzed using proportional hazards regression models. Diabetes was analyzed as reported at baseline and as a time dependent variable using information obtained during follow-up. After adjustment for BMI, waist:hip ratio, and other covariates, the relative risk (RR) for women with diabetes versus women without diabetes was 1.43 [95% confidence interval (CI), 0.98-2.1]. The diabetes association was confined to women in the upper two BMI quintiles (RR, 1.47; 95% CI, 0.98-2.20), but a formal test of interaction was not statistically significant. Analyses that included diabetes ascertained at baseline and at follow-up gave similar results; the diabetes-associated RR in the higher BMI strata was 1.64 (95% CI, 1.16-2.31). We conclude that after adjustment for other risk factors, diabetes is associated with a modestly increased risk for endometrial cancer among women in this cohort. PMID- 11401911 TI - Hepatitis B, aflatoxin B(1), and p53 codon 249 mutation in hepatocellular carcinomas from Guangxi, People's Republic of China, and a meta-analysis of existing studies. AB - The incidence of hepatocellular carcinomas (HCC) varies widely worldwide, with some of the highest incidence rates found in China. Chronic infection with the hepatitis B virus (HBV) and exposure to aflatoxins in foodstuffs are the main risk factors. A G to T transversion at codon 249 of the p53 gene (249(ser)) is commonly found in HCCs from patients in regions with dietary aflatoxin exposure. Because HBV infection is often endemic in high aflatoxin exposure areas, it is still unclear whether HBV acts as a confounder or as a synergistic partner in the development of the 249(ser) p53 mutation. Our report has two aims. First, we contribute data on HCCs from southern Guangxi, a high aflatoxin exposure area. Using DNA sequencing, we found that 36% (18 of 50) of tumors had a 249(ser) mutation. Also, 50% (30 of 60) were positive for p53 protein accumulation and 78% (28 of 36) were positive for HBV surface antigen, as detected by immunohistochemistry. Second, we present a meta-analysis, using our results along with those from 48 published studies, that examines the interrelationships among aflatoxin exposure, HBV infection, and p53 mutations in HCCs. We used a method that takes into account both within-study and study-to-study variability and found that the mean proportion of HCCs with the 249(ser) mutation was positively correlated with aflatoxin exposure (P = 0.0001). We found little evidence for an HBV-aflatoxin interaction modulating the presence of the p53 249(ser) mutation or any type of p53 mutation. PMID- 11401912 TI - Urinary estrogen metabolites and mammographic parenchymal patterns in postmenopausal women. AB - It has been hypothesized that women who metabolize their endogenous estrogens predominantly via 16(alpha)-hydroxylation rather than via 2-hydroxylation and, as a result, have a low ratio of 2-hydroxyestrone (2-OHE1):16(alpha)-hydroxyestrone (16(alpha)-OHE1) are at an increased risk of breast cancer. Epidemiological evidence in support of this hypothesis is scarce and mostly based on measurements made after the onset of the disease. To gain insight into the role of these metabolites in the etiology of breast cancer, we assessed their relationship with high-density Wolfe mammographic parenchymal patterns (P2/DY), a recognized indicator of risk of this tumor. The study was nested within a large cross sectional survey on determinants of mammographic patterns carried out in a population-based breast screening program in Northern Greece. Urinary levels of 2 OHE1 and 16(alpha)-OHE1 were measured in a random sample of 70 postmenopausal women with P2/DY mammographic patterns and in a random sample of 70 women with N1 mammographic patterns, individually matched to the P2/DY women on year of birth, years since menopause and date of urine collection. Women with a P2/DY pattern had, on average, 58% higher levels of 2-OHE1 (P = 0.002) and 15% higher levels of 16(alpha)-OHE1 (P = 0.37) than those with an N1 pattern. The ratio of 2 OHE1:16(alpha)-OHE1 was 35% higher (P = 0.005) in women with a P2/DY pattern. Women in the highest one-third of this ratio were six times more likely to have a P2/DY pattern than those in the lowest one-third after adjusting for potential confounders (prevalence odds ratio, 6.2; 95% CI, 1.7-22.9; test for linear trend, P = 0.002). These findings seem to suggest that a high, rather than a low, 2 OHE1:16(alpha)-OHE1 ratio may be associated with an increase in breast cancer risk at postmenopausal ages, unless the pathway through which estrogen metabolites may affect breast cancer risk is unrelated to mammographic parenchymal patterns. PMID- 11401913 TI - Polymorphic catechol-O-methyltransferase gene and breast cancer risk. AB - We examined 483 Finnish breast cancer cases and 482 population controls to determine the potential effect of catechol-O-methyltransferase (COMT) genotype in individual susceptibility to breast cancer. Odds ratios (ORs) and 95% confidence intervals (CIs) were estimated by unconditional logistic regression after adjustment for known or suspected risk factors for breast cancer. When studied separately by menopausal status, the COMT-L allele-containing genotypes were inversely associated with premenopausal breast cancer, especially with advanced stage of the disease (OR, 0.44; 95% CI, 0.22-0.87). Among postmenopausal women a similar decreased risk was seen for local carcinoma associated with the COMT-LL genotype (OR, 0.55; 95% CI, 0.31-0.98). The lowest breast cancer risk was seen in the postmenopausal women with the COMT-LL genotype and low body-mass index (30 months) use of estrogen (OR, 4.02; 95% CI, 1.13-14.3), or with the COMT-L allele-containing genotypes and early age (50 years of age (P = 0.05). Given the same duration of exposure, women had higher anti-HMdU aAbs and also reached peak levels at a lower cumulative smoking exposure (30 years) compared with male smokers (40 years). Subjects smoked an average of 28.9 +/- 0.81 CPD and initiated smoking at 17.2 +/- 0.33 (SE) years of age. Therefore, smokers who reported smoking for 30 years were typically <50 years old. Women 23 kb), in contrast to 55-61% of the brush extracts. PCR success rates for amplification of beta-globin gene fragments (268, 536, and 989 bp) were similar for cytobrush and mouthwash phenol-chloroform extracts (range, 94.4-100%). Also, we obtained high success rates in determining the number of CAG repeats in the androgen receptor gene, characterizing tetranucleotide microsatellites in six gene loci, and screening for mutations in the BRCA1/2 genes in a subset of phenol-chloroform DNA extracts. Relative to DNA extracted by phenol-chloroform from cytobrush samples, DNA extracted by NaOH had lower molecular weight, decreased PCR success rates for most assays performed, and unreliably high spectrophotometer readings for DNA yields. In conclusion, although DNA isolated from either mouthwash or cytobrush samples collected by mail from adults is adequate for a wide range of PCR-based assays, a single mouthwash sample provides substantially larger amounts and higher molecular weight DNA than two cytobrush samples. PMID- 11401921 TI - Whole genome amplification increases the efficiency and validity of buccal cell genotyping in pediatric populations. AB - The collection of buccal cells provides a noninvasive method for obtaining DNA for genetic studies. Here we report the results on buccal cell genotyping from our ongoing study of childhood leukemia in Northern California. We have collected buccal samples from children ranging in age from 4 months to 15 years using an interviewer- or nurse-administered protocol using a cytology brush. Initial results of the genotyping, including the glutathione S-transferase mu, glutathione S-transferase theta, NAD(P)H:quinone oxidoreductase, and methylenetetrahydrofolate reductase polymorphisms, were disappointing because many specimens contained little DNA, failed repeated attempts at PCR amplification, and produced unreliable results. Here we evaluate a solution to the problem that involves whole genome amplification using the improved primer extension preamplification methodology. Sixty cases of pediatric acute leukemia were studied; five PCR-based genotypes were attempted using buccal cell DNA and whole genome amplified (WGA) buccal DNA. Results were compared with genotyping results using DNA isolated from peripheral whole blood or bone marrow for each child. The standard buccal protocol failed to yield successful PCR reactions in 30-57% of specimens, whereas WGA-buccal was markedly more efficient (2-5% failed PCR). A success rate of 100% was achieved with one repeat test of the failed WGA PCR reactions. Misclassification of genotype was common for the glutathione S transferase theta marker using the standard buccal procedure. The WGA-buccal protocol, however, produced genotyping results fully concordant with the referent blood or bone marrow DNA results for all five loci. DNA yields were increased by WGA to allow for approximately 900 PCR reactions/brush. WGA is very useful for improving the efficiency and validity of PCR-based genotyping in pediatric populations. PMID- 11401922 TI - Feasibility of collecting buccal cell DNA by mail in a cohort study. AB - This study assessed the feasibility of obtaining buccal cell DNA by mail from participants in a large, community-based cohort study in Hawaii. Mouthwash collection kits were sent to a total of 355 randomly selected Japanese, Caucasian, and Hawaiian cohort members. Subjects were requested to swish 10 ml of mouthwash in their mouth for 60 s and expel it into a collection cup, which they mailed back to our laboratory. Half of the subjects were requested to collect a second sample. After up to two mailings and two reminder phone calls, two-thirds of the subjects returned a sample. The participation rate was lower for Hawaiians (59.0%) than for Caucasians (68.1%) and Japanese (76.3%). Participation was not affected by requesting two specimens. Participants did not differ from the total sample in terms of education and smoking status. The mean DNA yield was lower in females (41.7 microg) than males (53.4 microg) and in Japanese (37.8 microg) as compared with Hawaiians (51.9 microg) and Caucasians (54.5 microg). For subjects who returned two samples, the DNA yields were similar when both specimens were extracted in the same batch. All samples were successfully genotyped for polymorphisms in the CYP1A1, CYP2E1, GSTM1, GSTT1, and NQO1 genes by PCR-RFLP. From these and previous data, we conclude that, in situations where blood samples cannot be obtained, mail collection of mouthwash samples should be considered because it yields substantial amounts of high-quality genomic DNA for large numbers of study subjects. PMID- 11401923 TI - Epstein-Barr Virus and HLA-DPB1-*0301 in young adult Hodgkin's disease: evidence for inherited susceptibility to Epstein-Barr Virus in cases that are EBV(+ve). AB - Cases of Hodgkin's disease (HD) may be distinguished by whether they do [EBV positive ((+ve)) cases] or do not [EBV-negative ((-ve)) cases] have evidence of EBV DNA in the Reed-Sternberg cells. Only one study has attempted to distinguish epidemiological risk factors for EBV(+ve) and EBV(-ve) HD, and none have compared inherited susceptibility. The present study involves a population-based case series of HD, diagnosed in patients between 16-24 years of age in the United Kingdom (n = 118), of whom 87% were classified by EBV status (EBV(+ve), 19, EBV( ve), 84). History of infectious illness, EBV antibody titers, and HLA-DPB1 type have been compared in EBV(+ve) and EBV(-ve) cases. Reported infectious mononucleosis was more frequent in EBV(+ve) cases (odds ratio (OR), 5.10; 95% confidence interval (CI), 1.12-24.4). EBV antibody titers to viral capsid antigen were significantly higher in EBV(+ve) cases (P for trend = 0.02). Higher proportions of EBV(+ve) (43%) than EBV(-ve) (31%) cases typed positive for HLA DPB1*0301, but this was not statistically significant; the association of infectious mononucleosis with EBV(+ve) cases was stronger in this HLA subgroup (OR, 17.1; 95%CI, 1.06-1177) than in other cases (OR, 1.24; 95% CI, 0.02-15.4). Although these results are based on small numbers of HD cases, they provide suggestive evidence that the etiology of EBV(+ve) HD may involve inherited susceptibility to EBV. PMID- 11401924 TI - Association of genetic polymorphisms in UGT1A1 with breast cancer and plasma hormone levels. AB - UDP-glucuronosyltransferases (UGTs) catalyze the detoxification and the elimination of a large number of endogenous and exogenous compounds in the liver and extrahepatic tissues. One of the UGT1A family members, UGT1A1, is involved in estradiol metabolism and, therefore, represents a candidate gene in breast carcinogenesis. A common insertion/deletion polymorphism in the TATA-box of the promoter region of UGT1A1 results in decreased initiation of transcription. In a previous study, we found a positive association between the UGT1A1 low transcriptional alleles and premenopausal breast cancer risk in an African American population. In the present study, we sought to determine whether the low transcription UGT1A1 promoter allele, UGT1A1*28 [A(TA)(7)TAA], was associated with increased breast cancer risk among primarily Caucasian women in a nested case-control study within the Nurses' Health Study cohort. No significant association between the UGT1A1*28 [A(TA)(7)TAA] allele and breast cancer was observed. Compared with women homozygous for the UGT1A1*1 [A(TA)(6)TAA] allele, the relative risk was 0.80 (confidence interval, 0.49-1.29) for women homozygous for the UGT1A1*28 allele. The effect of the UGT1A1 genotype on plasma hormone levels in postmenopausal women not using hormone replacement was also evaluated, and overall, no significant differences in hormone levels by genotypes were observed. When restricted to women who had at least one UGT1A1*28 allele and a body mass index at blood draw of >27 kg/m(2), particularly in combination with the cytochrome p450c17alpha genotype, estrone and estradiol levels tended to vary by UGT1A1 genotypes. The results presented do not support a strong association between the UGT1A1 promoter polymorphism and the risk of breast cancer. PMID- 11401925 TI - Colonic mucosal concentrations of folate are accurately predicted by blood measurements of folate status among individuals ingesting physiologic quantities of folate. AB - Folate status is inversely related to the risk of colorectal cancer. Whether conventional blood measurements of folate status accurately reflect folate concentrations in the colorectal mucosa has been a controversial topic. This is an important issue because accurate measures of folate status in the colorectal mucosa are important for ascertaining the risk of colorectal cancer in epidemiological studies and for determining the effects of folate supplementation in clinical trials. We examined whether conventional blood measurements of folate and a more sensitive, inverse indicator of systemic folate status, serum homocysteine, accurately reflect folate concentrations in human colonic mucosa obtained by endoscopic biopsy. Study subjects (n = 20) were participants in a randomized trial that investigated the effect of folate supplementation (5 mg daily for 1 year) on provisional molecular markers of colon cancer. Blood samples and biopsies of normal rectosigmoid mucosa were obtained at baseline, at 6 months, and at 1 year. Serum, RBC, and colonic mucosal folate and serum homocysteine concentrations were determined. Colonic mucosal folate concentrations correlated directly with serum folate concentrators at each time point (r = 0.572-0.845; P < 0.015) and with RBC folate concentrations at 6 months and 1 year (r = 0.747-0.771; P < 0.001). Colonic mucosal folate concentrations correlated inversely with serum homocysteine concentrations at each time point (r = -0.622-0.666; P < 0.008). Systemic measures of folate status did not correlate with colonic mucosal folate concentrations among individuals receiving supplemental folate. Our observations indicate that colonic mucosal concentrations of folate may be predicted accurately by blood measurements of folate status only among individuals not ingesting supraphysiological quantities of folate. PMID- 11401926 TI - Cholecystokinin and gastrin levels are not elevated in human pancreatic adenocarcinoma. PMID- 11401927 TI - In-hospital initiation of lipid-lowering therapy for patients with coronary heart disease: the time is now. PMID- 11401928 TI - Correlation of cerebral metabolites with clinical outcome among patients with severe congestive heart failure. PMID- 11401929 TI - Go with the flow. PMID- 11401930 TI - Mobilization of endothelial progenitor cells in patients with acute myocardial infarction. AB - BACKGROUND: Endothelial progenitor cells (EPCs) circulate in adult peripheral blood (PB) and contribute to neovascularization. However, little is known regarding whether EPCs and their putative precursor, CD34-positive mononuclear cells (MNC(CD34+)), are mobilized into PB in acute ischemic events in humans. METHODS AND RESULTS: Flow cytometry revealed that circulating MNC(CD34+) counts significantly increased in patients with acute myocardial infarction (n=16), peaking on day 7 after onset, whereas they were unchanged in control subjects (n=8) who had no evidence of cardiac ischemia. During culture, PB-MNCs formed multiple cell clusters, and EPC-like attaching cells with endothelial cell lineage markers (CD31, vascular endothelial cadherin, and kinase insert domain receptor) sprouted from clusters. In patients with acute myocardial infarction, more cell clusters and EPCs developed from cultured PB-MNCs obtained on day 7 than those on day 1. Plasma levels of vascular endothelial growth factor significantly increased, peaking on day 7, and they positively correlated with circulating MNC(CD34+) counts (r=0.35, P=0.01). CONCLUSIONS: This is the first clinical demonstration showing that lineage-committed EPCs and MNC(CD34+), their putative precursors, are mobilized during an acute ischemic event in humans. PMID- 11401931 TI - Visualization of discrete microinfarction after percutaneous coronary intervention associated with mild creatine kinase-MB elevation. AB - BACKGROUND: Mild elevations in creatine kinase-MB (CK-MB) are common after successful percutaneous coronary interventions and are associated with future adverse cardiac events. The mechanism for CK-MB release remains unclear. A new contrast-enhanced MRI technique allows direct visualization of myonecrosis. METHODS AND RESULTS: Fourteen patients without prior infarction underwent cine and contrast-enhanced MRI after successful coronary stenting; 9 patients had procedure-related CK-MB elevation, and 5 did not (negative controls). The mean age of all patients was 61 years, 36% had diabetes, 43% had multivessel coronary artery disease, and all had a normal ejection fraction. Twelve patients (86%) received an intravenous glycoprotein IIb/IIIa inhibitor; none underwent atherectomy, and all had final TIMI 3 flow. Of the 9 patients with CK-MB elevation, 5 had a minor side branch occlusion during stenting, 2 had transient ECG changes, and none developed Q-waves. The median CK-MB was 21 ng/mL (range, 12 to 93 ng/mL), which is 2.3x the upper limit of normal. Contrast-enhanced MRI demonstrated discrete regions of hyperenhancement within the target vessel perfusion territory in all 9 patients. Only one developed a new wall motion abnormality. The median estimated mass of myonecrosis was 2.0 g (range, 0.7 to 12.2 g), or 1.5% of left ventricular mass (range, 0.4% to 6.0%). Hyperenhancement persisted in 5 of the 6 who underwent a repeat MRI at 3 to 12 months. No control patient had hyperenhancement. CONCLUSIONS: Contrast-enhanced MRI provides an anatomical correlate to biochemical evidence of procedure-related myocardial injury, despite the lack of ECG changes or wall motion abnormalities. Mild elevation of CK-MB after percutaneous coronary intervention is the result of discrete microinfarction. PMID- 11401932 TI - Prognostic significance of cerebral metabolic abnormalities in patients with congestive heart failure. AB - BACKGROUND: Cerebral metabolic abnormalities were proposed as a potential marker of disease severity in congestive heart failure (CHF), but their prognostic significance remains uncertain. METHODS AND RESULTS: We investigated the prognostic value of cerebral metabolic abnormalities in 130 consecutive patients with advanced CHF (100 men aged 42.6+/-11.9 years; left ventricular ejection fraction, 22.2+/-6.2%). Proton magnetic resonance spectroscopy data were obtained from localized regions ( approximately 8 mL) of the occipital gray matter and the parietal white matter. The primary end point was the occurrence of death after the proton magnetic resonance spectroscopy. During follow-up (18.5+/-14.4 months), 21 patients died and 15 underwent urgent heart transplantation. In the Cox proportional model, occipital metabolites (N-acetylaspartate, creatine, choline, and myoinositol), parietal N-acetylaspartate level, and the duration of CHF symptoms (>12 months) were validated as univariate predictors of death. In multivariate Cox analyses, however, the occipital N-acetylaspartate level was an independent predictor of death (hazard ratio, 0.52; 95% CI, 0.41 to 0.67; P<0.001). An analysis with respect to the combined end point of death or urgent transplantation showed similar results. The best cutoff value (9.0 mmol/kg) for occipital N-acetylaspartate level had 75% sensitivity and 67% specificity to predict mortality. CONCLUSIONS: The occipital N-acetylaspartate level is a powerful and independent predictor of CHF mortality, suggesting that cerebral metabolic abnormalities may be used as a new prognostic marker in the assessment of patients with CHF. PMID- 11401933 TI - Low circulating vitamin B(6) is associated with elevation of the inflammation marker C-reactive protein independently of plasma homocysteine levels. AB - BACKGROUND: Lower vitamin B(6) concentrations are reported to confer an increased and independent risk for cardiovascular disease (CVD). The mechanism underlying this relationship, however, remains to be defined. Other diseases, such as rheumatoid arthritis, are associated with reduced vitamin B(6) levels. Despite a clear distinction in pathophysiology, inflammatory reaction may be the major link between these diseases. We hypothesized a relationship between pyridoxal 5' phosphate (PLP), the active form of vitamin B(6), and the marker of inflammation C-reactive protein (CRP). We also evaluated whether total plasma homocysteine (tHcy), a well-defined risk factor for CVD and a major determinant of plasma PLP levels, had a possible role as a mediator of this hypothesized relationship. METHODS AND RESULTS: Data from 891 participants from the population-based Framingham Heart Study cohort were analyzed. Subjects were divided into 2 groups according to normal or elevated CRP values: group 1, CRP <6 mg/L; group 2, CRP >/=6 mg/L. Plasma PLP levels were substantially lower in group 2 than in group 1 (mean values in group 2, 36.5 nmol/L versus 55.8 nmol/L in group 1, P<0.001). In a multiple logistic regression model adjusted for tHcy, the association of PLP with CRP remained highly significant (P=0.003). CONCLUSIONS: Low plasma PLP is associated with higher CRP levels independently of tHcy. This observation may reflect a vitamin B(6) utilization in the presence of an underlying inflammatory process and represent a possible mechanism to explain the decreased vitamin B(6) levels in CVD. PMID- 11401934 TI - Select flavonoids and whole juice from purple grapes inhibit platelet function and enhance nitric oxide release. AB - BACKGROUND: Moderate red wine consumption is inversely associated with coronary ischemia, and both red wine and purple grape juice (PGJ) contain flavonoids with antioxidant and antiplatelet properties believed to be protective against cardiovascular events. Acute cardiac events are also associated with decreased platelet-derived nitric oxide (NO) release. In this study, the effects of PGJ and PGJ-derived flavonoids on platelet function and platelet NO production were determined. METHODS AND RESULTS: Incubation of platelets with dilute PGJ led to inhibition of aggregation, enhanced release of platelet-derived NO, and decreased superoxide production. To confirm the in vivo relevance of these findings, 20 healthy subjects consumed 7 mL. kg(-1). d(-1) of PGJ for 14 days. Platelet aggregation was inhibited after PGJ supplementation, platelet-derived NO production increased from 3.5+/-1.2 to 6.0+/-1.5 pmol/10(8) platelets, and superoxide release decreased from 29.5+/-5.0 to 19.2+/-3.1 arbitrary units (P<0.007 and P<0.05, respectively). alpha-Tocopherol levels increased significantly after PGJ consumption (from 15.6+/-0.7 to 17.6+/-0.9 micromol/L; P<0.009), and the plasma protein-independent antioxidant activity increased by 50.0% (P<0.05). Last, incubation of platelets with select flavonoid fractions isolated from PGJ consistently attenuated superoxide levels but had variable effects on whole-blood aggregation, platelet aggregation, and NO release. CONCLUSIONS: Both in vitro incubation and oral supplementation with PGJ decrease platelet aggregation, increase platelet-derived NO release, and decrease superoxide production. These findings may be a result of antioxidant-sparing and/or direct effects of select flavonoids found in PGJ. The suppression of platelet-mediated thrombosis represents a potential mechanism for the beneficial effects of purple grape products, independent of alcohol consumption, in cardiovascular disease. PMID- 11401935 TI - Iron chelation improves endothelial function in patients with coronary artery disease. AB - BACKGROUND: Some epidemiological studies have shown that increased iron stores are associated with increased cardiovascular events. Redox-active iron may contribute to lipid peroxidation, endothelial cell activation, and generation of reactive oxygen species (especially hydroxyl radical, via Fenton chemistry). Increased oxidative stress is associated with impaired action of endothelium derived nitric oxide in patients with atherosclerosis. METHODS AND RESULTS: To test the hypothesis that reducing vascular iron stores would reverse endothelial dysfunction, we examined the effects of the iron chelator deferoxamine (500 mg intra-arterially over 1 hour) on vasomotor function in forearm resistance vessels of patients with coronary artery disease by venous occlusion plethysmography. Patients with coronary artery disease had impaired endothelium-dependent vasodilation in response to methacholine compared with healthy control subjects (P<0.001). Deferoxamine infusion decreased serum iron levels (P<0.001). Deferoxamine improved the blood flow response to methacholine in patients with coronary artery disease (P<0.01 by 2-way repeated-measures ANOVA) but had no effect on the response to sodium nitroprusside. In normal volunteers, deferoxamine had no effect on the response to methacholine. The nitric oxide synthase inhibitor N(G)-monomethyl-L-arginine abolished augmentation of the methacholine response associated with deferoxamine. The hydroxyl radical scavenger mannitol had no effect on the methacholine response. CONCLUSIONS: Deferoxamine improved nitric oxide-mediated, endothelium-dependent vasodilation in patients with coronary artery disease. These results suggest that iron availability contributes to impaired nitric oxide action in atherosclerosis. PMID- 11401936 TI - Triggering myocardial infarction by marijuana. AB - BACKGROUND: Marijuana use in the age group prone to coronary artery disease is higher than it was in the past. Smoking marijuana is known to have hemodynamic consequences, including a dose-dependent increase in heart rate, supine hypertension, and postural hypotension; however, whether it can trigger the onset of myocardial infarction is unknown. METHODS AND RESULTS: In the Determinants of Myocardial Infarction Onset Study, we interviewed 3882 patients (1258 women) with acute myocardial infarction an average of 4 days after infarction onset. We used the case-crossover study design to compare the reported use of marijuana in the hour preceding symptoms of myocardial infarction onset to its expected frequency using self-matched control data. Of the 3882 patients, 124 (3.2%) reported smoking marijuana in the prior year, 37 within 24 hours and 9 within 1 hour of myocardial infarction symptoms. Compared with nonusers, marijuana users were more likely to be men (94% versus 67%, P<0.001), current cigarette smokers (68% versus 32%, P<0.001), and obese (43% versus 32%, P=0.008). They were less likely to have a history of angina (12% versus 25%, P<0.001) or hypertension (30% versus 44%, P=0.002). The risk of myocardial infarction onset was elevated 4.8 times over baseline (95% confidence interval, 2.4 to 9.5) in the 60 minutes after marijuana use. The elevated risk rapidly decreased thereafter. CONCLUSIONS: Smoking marijuana is a rare trigger of acute myocardial infarction. Understanding the mechanism through which marijuana causes infarction may provide insight into the triggering of myocardial infarction by this and other, more common stressors. PMID- 11401937 TI - Increased particulate air pollution and the triggering of myocardial infarction. AB - BACKGROUND: Elevated concentrations of ambient particulate air pollution have been associated with increased hospital admissions for cardiovascular disease. Whether high concentrations of ambient particles can trigger the onset of acute myocardial infarction (MI), however, remains unknown. METHODS AND RESULTS: We interviewed 772 patients with MI in the greater Boston area between January 1995 and May 1996 as part of the Determinants of Myocardial Infarction Onset Study. Hourly concentrations of particle mass <2.5 microm (PM(2.5)), carbon black, and gaseous air pollutants were measured. A case-crossover approach was used to analyze the data for evidence of triggering. The risk of MI onset increased in association with elevated concentrations of fine particles in the previous 2-hour period. In addition, a delayed response associated with 24-hour average exposure 1 day before the onset of symptoms was observed. Multivariate analyses considering both time windows jointly revealed an estimated odds ratio of 1.48 associated with an increase of 25 microg/m(3) PM(2.5) during a 2-hour period before the onset and an odds ratio of 1.69 for an increase of 20 microg/m(3) PM(2.5) in the 24-hour period 1 day before the onset (95% CIs 1.09, 2.02 and 1.13, 2.34, respectively). CONCLUSIONS: The present study suggests that elevated concentrations of fine particles in the air may transiently elevate the risk of MIs within a few hours and 1 day after exposure. Further studies in other locations are needed to clarify the importance of this potentially preventable trigger of MI. PMID- 11401938 TI - Intracoronary stenting and angiographic results: strut thickness effect on restenosis outcome (ISAR-STEREO) trial. AB - BACKGROUND: Increased thrombogenicity and smooth muscle cell proliferative response induced by the metal struts compromise the advantages of coronary stenting. The objective of this randomized, multicenter study was to assess whether a reduced strut thickness of coronary stents is associated with improved follow-up angiographic and clinical results. METHODS AND RESULTS: A total of 651 patients with coronary lesions situated in native vessels >2.8 mm in diameter were randomly assigned to receive 1 of 2 commercially available stents of comparable design but different thickness: 326 patients to the thin-strut stent (strut thickness of 50 microm) and 325 patients to the thick-strut stent (strut thickness of 140 microm). The primary end point was the angiographic restenosis (>/=50% diameter stenosis at follow-up angiography). Secondary end points were the incidence of reinterventions due to restenosis-induced ischemia and the combined rate of death and myocardial infarctions at 1 year. The incidence of angiographic restenosis was 15.0% in the thin-strut group and 25.8% in the thick strut group (relative risk, 0.58; 95% CI, 0.39 to 0.87; P=0.003). Clinical restenosis was also significantly reduced, with a reintervention rate of 8.6% among thin-strut patients and 13.8% among thick-strut patients (relative risk, 0.62; 95% CI, 0.39 to 0.99; P=0.03). No difference was observed in the combined 1 year rate of death and myocardial infarction. CONCLUSIONS: The use of a thinner strut device is associated with a significant reduction of angiographic and clinical restenosis after coronary artery stenting. These findings may have relevant implications for the currently most widely used percutaneous coronary intervention. PMID- 11401939 TI - Autosomal recessive catecholamine- or exercise-induced polymorphic ventricular tachycardia: clinical features and assignment of the disease gene to chromosome 1p13-21. AB - BACKGROUND: Catecholaminergic polymorphic ventricular tachycardia (PVT) is characterized by episodes of syncope, seizures, or sudden death in response to physiological or emotional stress. In 2 families with autosomal dominant inheritance, the disease gene was mapped to chromosome 1q42-43. The objectives of this study were to characterize the clinical features of the disease in a Bedouin tribe from Israel and to map the disease gene. METHODS AND RESULTS: In this Bedouin tribe, 9 children (age, 7+/-4 years) from 7 related families have died suddenly during the past decade, and 12 other children suffered from recurrent syncope and seizures starting at the age of 6+/-3 years. Parents of affected individuals were asymptomatic and were all related (first-, second-, or third degree cousins). Segregation analysis suggested autosomal recessive inheritance. All 12 symptomatic patients and 1 asymptomatic sibling (mean age, 13+/-7 years) were found to have a relative resting bradycardia (64+/-13 bpm, versus 93+/-12 bpm in the unaffected siblings), as well as PVT induced by treadmill or isoproterenol infusion and appearing at a mean sinus rate of 110+/-10 bpm. Patients responded favorably to treatment with beta-blockers. A genome-wide search using polymorphic DNA markers mapped the disease locus to a 16-megabase interval on chromosome 1p13-21. A maximal lod score of 8.24 was obtained with D1S189 at theta=0.00. Sequencing of KCND3, a gene that encodes an I(tO) potassium channel transporter, did not reveal any significant sequence alterations. CONCLUSIONS: This unique form of autosomal recessive PVT affects young children and may be lethal if left untreated. Linkage analysis maps this disorder to chromosome 1p13-21. PMID- 11401940 TI - Reduction in stroke with gemfibrozil in men with coronary heart disease and low HDL cholesterol: The Veterans Affairs HDL Intervention Trial (VA-HIT). AB - BACKGROUND: A low level of HDL cholesterol has been identified as a risk factor for stroke in observational studies. METHODS AND RESULTS: Our objective was to determine whether treatment aimed at raising HDL cholesterol and lowering triglycerides reduces stroke in men with coronary heart disease and low levels of both HDL and LDL cholesterol. The study was a placebo-controlled, randomized trial conducted in 20 Veterans Affairs medical centers. A total of 2531 men with coronary heart disease, with mean HDL cholesterol 0.82 mmol/L (31.5 mg/dL) and mean LDL cholesterol 2.9 mmol/L (111 mg/dL), were randomized to gemfibrozil 1200 mg/d or placebo and were followed up for 5 years. Strokes were confirmed by a blinded adjudication committee. Relative risks were derived from Cox proportional hazards models. There were 134 confirmed strokes, 90% of which were ischemic. Seventy-six occurred in the placebo group (9 fatal) and 58 in the gemfibrozil group (3 fatal), for a relative risk reduction, adjusted for baseline variables, of 31% (95% CI, 2% to 52%, P=0.036). The reduction in risk was evident after 6 to 12 months. Patients with baseline HDL cholesterol below the median may have been more likely to benefit from treatment than those with higher HDL cholesterol. CONCLUSIONS: In men with coronary heart disease, low HDL cholesterol, and low LDL cholesterol, gemfibrozil reduces stroke incidence. PMID- 11401941 TI - Chlamydia pneumoniae infection does not induce or modify atherosclerosis in mice. AB - BACKGROUND: Seroepidemiological studies have linked Chlamydia pneumoniae (CP) to coronary heart disease, and recent experimental studies suggest that it may accelerate or even induce atherosclerosis. We therefore evaluated the effect of CP infection on atherosclerosis in atherosclerosis-prone apolipoprotein E knockout (apoE-KO) and wild-type C57BL/6J mice. METHODS AND RESULTS: Six- to 8 week-old female mice were infected intranasally with live CP and then fed a standard chow diet for 22 weeks. A subgroup of mice was reinfected 18 weeks after primary infection. Polymerase chain reaction analysis of lung tissue confirmed successful infection with CP, and ELISA assays demonstrated development of a humoral immune response. Despite this, no statistically significant differences in aortic atherosclerotic lesions were found between CP-infected and control apoE KO mice. Furthermore, CP infection did not induce atherosclerosis in C57BL/6J mice. CONCLUSIONS: CP does not induce atherosclerosis in wild-type mice and does not accelerate atherosclerosis in chow-fed apoE-KO mice. Further studies will be necessary to clarify the explanation for the seroepidemiological association between CP and coronary heart disease in humans. PMID- 11401942 TI - Dysfunctional regulation of endothelial nitric oxide synthase (eNOS) expression in response to exercise in mice lacking one eNOS gene. AB - BACKGROUND: Previous data suggest that 1 endothelial NO synthase (eNOS) gene is sufficient to allow normal expression and function of eNOS under basal conditions. We hypothesized that this might not hold true for conditions known to increase eNOS gene expression, such as exercise. METHODS AND RESULTS: Male mice heterozygous for a disruption of the eNOS gene (eNOS(+/)(-)) and normal C56Bl/6J mice (eNOS(+/+)), 3 to 4 months of age, underwent exercise training for 3 weeks. Nontrained mice were exposed to the exercise environment (noise and vibration of the treadmill) without exercise for an identical period. In eNOS(+/+) mice (n=7), exercise increased aortic eNOS protein expression by 3.4+/-0.4-fold (P<0.002). This was associated with a greater vascular cGMP accumulation on stimulation with acetylcholine (P<0.05). Furthermore, exercise training increased eNOS mRNA (1.78+/-0.4-fold) and protein (1.76+/-0.17-fold) in left ventricular tissue, as determined by competitive reverse transcription-polymerase chain reaction and Western analysis (P<0.05 for both). In striking contrast, exercise had no effect on aortic eNOS expression and cGMP accumulation in eNOS(+/)(-) mice (P>0.05). Thus, although eNOS expression appears to be normal in eNOS(+/)(-) mice under basal conditions, these mice are unable to increase eNOS expression during exercise. CONCLUSIONS: These findings show that regulation of eNOS expression during exercise requires the presence of both alleles of the gene and may have implications for conditions in which polymorphisms of eNOS are present in only 1 allele in humans. These individuals may have a normal vascular reactivity under basal conditions but may be unable to adapt their vascular reactivity in response to exercise training. PMID- 11401943 TI - Combined angiotensin II receptor antagonism and angiotensin-converting enzyme inhibition further attenuates postinfarction left ventricular remodeling. AB - BACKGROUND: ACE inhibition (ACEI) attenuates post-myocardial infarction (MI) LV remodeling, but the effects of angiotensin II type 1 receptor (AT(1)) antagonism alone or in combination with ACEI are unclear. Accordingly, we investigated the effects of AT(1) antagonism, ACEI, and their combination in a well-characterized ovine postinfarction model. METHODS AND RESULTS: Beginning 2 days after transmural anteroapical MI, 62 sheep were treated with 1 of 5 treatment regimens: no therapy (control, n=12), standard-dose ACEI (sACEI; ramipril 10 mg/d, n=14), high-dose ACEI (hACEI; ramipril 20 mg/d, n=8), AT(1) blockade (losartan 50 mg/d, n=13), and combination therapy with sACEI+AT(1) blockade (CT; ramipril 10 mg/d+losartan 50 mg/d, n=15). MRI was performed before and 8 weeks after MI to quantify changes in LV end-diastolic and end-systolic volume indices (DeltaEDVI, DeltaESVI) and ejection fraction (DeltaEF). Change in regional percent intramyocardial circumferential shortening in noninfarcted segments adjacent to the infarct (Adj Delta%S) was measured by tagged MRI. CT resulted in the most marked blunting of LV remodeling: DeltaESVI (+1.0+/-0.4, +0.7+/-0.4, +0.6+/-0.3, +0.9+/-0.5, and +0.4+/-0.2* mL/kg); DeltaEDVI (+0.9+/-0.4, +0.7+/-0.5, +0.6+/ 0.5, +0.9+/-0.5, and +0.4+/-0.3 mL/kg); DeltaEF (-24+/-7, -18+/-6, -14+/-7, -18+/ 10, and -11+/-9* %); and Adj Delta%S (-8+/-4, -7+/-3, -5+/-3, -5+/-3, and -2+/-3* %) for Control, sACEI, hACEI, AT(1) blockade, and CT, respectively (*P<0.04 versus sACEI, AT(1) blockade, and control; P<0.05 versus control; P<0.002 versus AT(1) blockade and control). EDVI and ESVI at 8 weeks after MI were smallest with CT (P<0.02 versus all). CONCLUSIONS: Combination therapy with sACEI+AT(1) blockade shows promise in attenuating postinfarction LV remodeling but was not clearly superior to hACEI in the present study. PMID- 11401944 TI - Phase 2 early afterdepolarization as a trigger of polymorphic ventricular tachycardia in acquired long-QT syndrome : direct evidence from intracellular recordings in the intact left ventricular wall. AB - BACKGROUND: This study examined the role of phase 2 early afterdepolarization (EAD) in producing a trigger to initiate torsade de pointes (TdP) with QT prolongation induced by dl-sotalol and azimilide. The contribution of transmural dispersion of repolarization (TDR) to transmural propagation of EAD and the maintenance of TdP was also evaluated. METHODS AND RESULTS: Transmembrane action potentials from epicardium, midmyocardium, and endocardium were recorded simultaneously, together with a transmural ECG, in arterially perfused canine and rabbit left ventricular preparations. dl-Sotalol preferentially prolonged action potential duration (APD) in M cells dose-dependently (1 to 100 micromol/L), leading to QT prolongation and an increase in TDR. Azimilide, however, significantly prolonged APD and QT interval at concentrations from 0.1 to 10 micromol/L but shortened them at 30 micromol/L. Unlike dl-sotalol, azimilide (>3 micromol/L) increased epicardial APD markedly, causing a diminished TDR. Although both dl-sotalol and azimilide rarely induced EADs in canine left ventricles, they produced frequent EADs in rabbits, in which more pronounced QT prolongation was seen. An increase in TDR by dl-sotalol facilitated transmural propagation of EADs that initiated multiple episodes of spontaneous TdP in 3 of 6 rabbit left ventricles. Of note, although azimilide (3 to 10 micromol/L) increased APD more than dl-sotalol, its EADs often failed to propagate transmurally, probably because of a diminished TDR. CONCLUSIONS: This study provides the first direct evidence from intracellular action potential recordings that phase 2 EAD can be generated from intact ventricular wall and produce a trigger to initiate the onset of TdP under QT prolongation. PMID- 11401945 TI - Assessment of global atrial fibrillation organization to optimize timing of atrial defibrillation. AB - BACKGROUND: We hypothesized that frequency domain analysis of a wide bipolar interatrial electrogram describes the global organization of atrial fibrillation (AF) and should vary over time. By timing shocks to periods of high organization of AF, cardioversion efficacy should improve. METHODS AND RESULTS: A total of 15 dogs (weight, 28.2+/-3.4 kg) were rapidly paced for 48 to 72 hours to induce AF. Coil electrodes with a surface area of 1.80 cm(2) were then placed in the left and right atria to form a wide bipole. Wide bipolar electrograms were digitally filtered, and a fast Fourier transform was performed over a sliding 2-s window every 0.5 s. The organization index (OI) was calculated as the ratio of the area of the dominant peak and its harmonics to the total area of the magnitude spectrum. The atrial defibrillation threshold (ADFT(50)) was determined using a 3 ms/3-ms biphasic shock and an up-down-up protocol. Additional shocks with higher and lower energies were delivered in a random sequence to develop a distribution curve. The OI varied over time, with a mean of 0.42+/-0.03, a maximum of 0.65+/ 0.07, and a minimum of 0.20+/-0.06. The OI changed rapidly, with durations of high organization (OI>0.5) ranging from 1 to 5 s. The ADFT(50) for QRS complex synchronized shocks was 183+/-56 V, versus 142+/-49 V for shocks synchronized to an OI>0.5 (P<0.001). The distribution curve shifted leftward when shocks were synchronized to an OI>0.5. CONCLUSIONS: AF signals show a high degree of variability. Shock efficacy is increased when shocks are delivered during periods of high AF organization as determined by the OI method. PMID- 11401946 TI - Fibrinolysis for acute myocardial infarction: current status and new horizons for pharmacological reperfusion, part 1. PMID- 11401947 TI - Images in cardiovascular medicine: Rare cause of cardiomyopathy. PMID- 11401948 TI - Noninvasive assessment of pseudo-pulmonary artery sling by echocardiography and computerized tomography. PMID- 11401949 TI - Vitamin C and coronary microcirculation. PMID- 11401950 TI - Beta-blockade in chronic heart failure. PMID- 11401951 TI - Sildenafil (Viagra) prolongs cardiac repolarization by blocking the rapid component of the delayed rectifier potassium current. PMID- 11401952 TI - Continuous positive airway pressure in patients with congestive heart failure and Cheyne-Stokes respiration with central sleep apnea. PMID- 11401953 TI - Beta-blocker trials seem to be in conflict. PMID- 11401954 TI - Tuberculosis: latency and reactivation. PMID- 11401955 TI - Differential gene expression from two transcriptional units in the cag pathogenicity island of Helicobacter pylori. AB - Infection with Helicobacter pylori strains containing the cag Pathogenicity Island (cag PAI) is strongly correlated with the development of severe gastric disease, including gastric and duodenal ulceration, mucosa-associated lymphoid tissue lymphoma, and gastric carcinoma. Although in vitro studies have demonstrated that the expression of genes within the cag PAI leads to the activation of a strong host inflammatory response, the functions of most cag gene products and how they work in concert to promote an immunological response are unknown. We developed a transcriptional reporter that utilizes urease activity and in which nine putative regulatory sequences from the cag PAI were fused to the H. pylori ureB gene. These fusions were introduced in single copies onto the H. pylori chromosome without disruption of the cag PAI. Our analysis indicated that while each regulatory region confers a reproducible amount of promoter activity under laboratory conditions, they differ widely in levels of expression. Transcription initiating upstream of cag15 and upstream of cag21 is induced when the respective fusion strains are cocultured with an epithelial cell monolayer. Results of mouse colonization experiments with an H. pylori strain carrying the cag15-ureB fusion suggested that this putative regulatory region appears to be induced in vivo, demonstrating the importance of the urease reporter as a significant development toward identifying in vivo-induced gene expression in H. pylori. PMID- 11401956 TI - Diepitopic construct of functionally and epitopically complementary peptides enhances immunogenicity, reactivity with glucosyltransferase, and protection from dental caries. AB - Coimmunization with peptide constructs from catalytic (CAT) and glucan-binding (GLU) domains of glucosyltransferase (GTF) of mutans streptococci has resulted in enhanced levels of antibody to the CAT construct and to GTF. We designed and synthesized a diepitopic construct (CAT-GLU) containing two copies of both CAT (B epitope only) and GLU (B and T epitope) peptides. The immunogenicity of this diepitopic construct was compared with that of individual CAT and GLU constructs by immunizing groups of Sprague-Dawley rats subcutaneously in the salivary gland vicinity with the CAT-GLU, CAT, or GLU construct or by treating rats by sham immunization. Levels of serum immunoglobulin G (IgG) antibody to GTF or CAT in the CAT-GLU group were significantly greater than in GLU- or CAT-immunized groups. Immunization with CAT-GLU was compared to coimmunization with a mixture of CAT and GLU in a second rodent experiment under a similar protocol. CAT-GLU immunization resulted in serum IgG and salivary IgA responses to GTF and CAT which were greater than after coimmunization. Immunization with the diepitopic construct and communization with CAT and GLU constructs showed proliferation of T lymphocytes to GTF. Immunization with either the CAT or GLU construct has been shown to elicit significant protection in a rodent dental caries model. Similarly in this study, the enhanced response to GTF after immunization with the CAT-GLU construct resulted in protective effects on dental caries. Therefore, the CAT-GLU diepitopic construct can be a potentially important antigen for a caries vaccine, giving rise to greater immune response than after immunization with CAT, GLU, or a mixture of the two. PMID- 11401957 TI - Whole-cell but not acellular pertussis vaccines induce convulsive activity in mice: evidence of a role for toxin-induced interleukin-1beta in a new murine model for analysis of neuronal side effects of vaccination. AB - Immunization with the whole-cell pertussis vaccine (Pw), while effective at preventing whooping cough in infants, has been associated with local, systemic, and neuronal reactions, including fevers and convulsions in children. In contrast, the new acellular pertussis vaccines (Pa) have a considerably improved safety profile. The lack of an appropriate animal model has restricted investigations into the mechanisms by which neurological reactions are induced by vaccination. Here we describe a novel murine model wherein seizure-like behavioral changes are induced following parenteral administration of Pw. The proinflammatory cytokine interleukin-beta (IL-1beta), production of which has been associated with many neurodegenerative conditions, was significantly increased in the hippocampus and hypothalamus of vaccinated animals. Accompanying this change was a decrease in release of the inhibitory neurotransmitters gamma aminobutyric acid and adenosine in the hippocampus. Seizure-like behavioral changes were significantly reduced following inhibition of IL-1beta production by the administration of an inhibitor of IL-1beta-converting enzyme and were almost completely abrogated in IL-1 receptor type I knockout mice. These results suggest a causal relationship between IL-1beta induction and convulsive behavior following Pw vaccination. Significantly, Pa neither increased IL-1beta nor induced behavioral changes in mice, but did induce the anti-inflammatory cytokine IL-10. In contrast, administration of active pertussis toxin and lipopolysaccharide, residual in Pw but absent from Pa, also induced convulsive activity. Our findings provide the first direct evidence of an immunological basis for pertussis vaccine reactogenicity and suggest that active bacterial toxins are responsible for the neurologic disturbances observed in children immunized with Pw. PMID- 11401958 TI - Characterization of Haemophilus ducreyi-specific T-cell lines from lesions of experimentally infected human subjects. AB - Haemophilus ducreyi is the etiologic agent of chancroid, a sexually transmitted genital ulcer disease that facilitates the transmission of human immunodeficiency virus. In the human model of infection, the histopathology of infected sites in part resembles a delayed-type hypersensitivity (DTH) response. In this study, T cells were isolated from skin biopsy specimens obtained from 24 subjects who were infected for 7 to 14 days. One clone and 12 lines that responded to H. ducreyi antigens were obtained from 12 of the subjects. Fluorescence-activated cell sorter analysis showed that the antigen-responsive lines and clone were predominantly CD3(+) and CD4(+). The lines and clone responded to H. ducreyi antigen in a dose-dependent manner and produced gamma interferon (IFN-gamma) alone or IFN-gamma and interleukin-10 (IL-10) but no IL-4 or IL-5 in response to H. ducreyi. Proliferation of T cells was dependent on the presence of autologous antigen-presenting cells. The lines showed little response to antigens prepared from other members of the Pasteurellaceae and responded to different fractions of H. ducreyi separated by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis. We conclude that T cells that recognize H. ducreyi antigens are recruited to sites experimentally infected with the organism. The lack of cross reactivity to the Pasteurellaceae and the response of the lines to different antigen fractions suggest that subjects are sensitized to H. ducreyi during the course of infection. PMID- 11401959 TI - Helicobacter hepaticus-induced colitis in interleukin-10-deficient mice: cytokine requirements for the induction and maintenance of intestinal inflammation. AB - We have previously shown that specific-pathogen-free interleukin-10 (IL-10) deficient (IL-10 KO) mice reconstituted with Helicobacter hepaticus develop severe colitis associated with a Th1-type cytokine response. In the present study, we formally demonstrate that IL-12 is crucial for disease induction, because mice deficient for both IL-10 and IL-12 p40 show no intestinal pathology following H. hepaticus infection. By using monoclonal antibodies (MAbs) to IL-12, gamma interferon (IFN-gamma), and tumor necrosis factor alpha (TNF-alpha), we have further analyzed the role of these cytokines in the maintenance of the Th1 response and inflammation in IL-10 KO mice with established H. hepaticus-induced colitis. Treatment of infected colitic IL-10 KO mice with anti-IL-12 p40 resulted in markedly reduced intestinal inflammation, colonic IFN-gamma, TNF-alpha, and inducible nitric oxide synthase (iNOS) mRNA levels, and H. hepaticus-specific IFN gamma secretion by mesenteric lymph node (MLN) cells compared to the findings in control MAb-treated mice. Moreover, the diminished pathology was associated with decreased numbers of colonic CD3(+) T cells and significantly reduced frequencies of Helicobacter-reactive CD4(+) Th1 cells in MLN. In contrast, anti-IFN-gamma and/or anti-TNF-alpha had no effect on intestinal inflammation in IL-10 KO mice with established colitis. Using IL-10/IFN-gamma double-deficient mice, we further show that IFN-gamma is not required for the development of colitis following H. hepaticus infection. MLN cells from infected IL-10/IFN-gamma KO animals secreted elevated amounts of IL-12 and TNF-alpha following bacterial antigen stimulation, indicating alternative pathways of disease induction. Taken together, our results demonstrate a crucial role for IL-12 in both inducing and sustaining intestinal inflammation through recruitment and maintenance of a pool of pathogenic Th1 cells. PMID- 11401960 TI - Calprotectin expression in vitro by oral epithelial cells confers resistance to infection by Porphyromonas gingivalis. AB - Calprotectin, an S100 calcium-binding protein with broad-spectrum antimicrobial activity in vitro, is expressed in neutrophils, monocytes, and gingival keratinocytes. In periodontitis, calprotectin appears upregulated and is detected at higher levels in gingival crevicular fluid and tissue specimens. How calprotectin contributes to the pathogenesis of periodontal diseases is unknown. To isolate the effects of calprotectin, a calprotectin-negative oral epithelial cell line was transfected with calprotectin genes to enable expression. Porphyromonas gingivalis was permitted to bind and invade transfected cells expressing calprotectin and sham transfectants. Rates of invasion into both cell lines were compared using the antibiotic protection assay. Transfected cells expressing calprotectin showed 40 to 50% fewer internalized P. gingivalis than sham transfectants. Similarly, binding to calprotectin expressing cells was reduced approximately twofold at all time points (15, 30, 45, and 60 min) as estimated by immunofluorescence analysis. Independent of invasion, however, prolonged exposure to P. gingivalis induced epithelial cell rounding and detachment from the substratum. These morphological changes were delayed, however, in cells expressing calprotectin. Using P. gingivalis protease-deficient mutants, we found that Arg-gingipain and Lys-gingipain contributed to epithelial cell rounding and detachment. In conclusion, expression of calprotectin appears to protect epithelial cells in culture against binding and invasion by P. gingivalis. In addition, cells expressing calprotectin are more resistant to detachment mediated by Arg-gingipain and Lys-gingipain. In periodontal disease, calprotectin may augment both the barrier protection and innate immune functions of the gingival epithelium to promote resistance to P. gingivalis infection. PMID- 11401961 TI - S-Fimbria-encoding determinant sfa(I) is located on pathogenicity island III(536) of uropathogenic Escherichia coli strain 536. AB - The sfa(I) determinant encoding the S-fimbrial adhesin of uropathogenic Escherichia coli strains was found to be located on a pathogenicity island of uropathogenic E. coli strain 536. This pathogenicity island, designated PAI III(536), is located at 5.6 min of the E. coli chromosome and covers a region of at least 37 kb between the tRNA locus thrW and yagU. As far as it has been determined, PAI III(536) also contains genes which code for components of a putative enterochelin siderophore system of E. coli and Salmonella spp. as well as for colicin V immunity. Several intact or nonfunctional mobility genes of bacteriophages and insertion sequence elements such as transposases and integrases are present on PAI III(536). The presence of known PAI III(536) sequences has been investigated in several wild-type E. coli isolates. The results demonstrate that the determinants of the members of the S-family of fimbrial adhesins may be located on a common pathogenicity island which, in E. coli strain 536, replaces a 40-kb DNA region which represents an E. coli K-12 specific genomic island. PMID- 11401962 TI - Motility and the polar flagellum are required for Aeromonas caviae adherence to HEp-2 cells. AB - Aeromonas caviae is increasingly being recognized as a cause of gastroenteritis, especially among the young. The adherence of aeromonads to human epithelial cells in vitro has been correlated with enteropathogenicity, but the mechanism is far from well understood. Initial investigations demonstrated that adherence of A. caviae to HEp-2 cells was significantly reduced by either pretreating bacterial cells with an antipolar flagellin antibody or by pretreating HEp-2 cells with partially purified flagella. To precisely define the role of the polar flagellum in aeromonad adherence, we isolated the A. caviae polar flagellin locus and identified five polar flagellar genes, in the order flaA, flaB, flaG, flaH, and flaJ. Each gene was inactivated using a kanamycin resistance cartridge that ensures the transcription of downstream genes, and the resulting mutants were tested for motility, flagellin expression, and adherence to HEp-2 cells. N terminal amino acid sequencing, mutant analysis, and Western blotting demonstrated that A. caviae has a complex flagellum filament composed of two flagellin subunits encoded by flaA and flaB. The predicted molecular mass of both flagellins was approximately 31,700 Da; however, their molecular mass estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was approximately 35,500 Da. This aberrant migration was thought to be due to their glycosylation, since the proteins were reactive in glycosyl group detection assays. Single mutations in either flaA or flaB did not result in loss of flagella but did result in decreased motility and adherence by approximately 50%. Mutation of flaH, flaJ, or both flagellin genes resulted in the complete loss of motility, flagellin expression, and adherence. However, mutation of flaG did not affect motility but did significantly reduce the level of adherence. Centrifugation of the flagellate mutants (flaA, flaB, and flaG) onto the cell monolayers did not increase adherence, whereas centrifugation of the aflagellate mutants (flaH, flaJ, and flaA flaB) increased adherence slightly. We conclude that maximum adherence of A. caviae to human epithelial cells in vitro requires motility and optimal flagellar function. PMID- 11401963 TI - Interleukin-6 enhances production of anti-OspC immunoglobulin G2b borreliacidal antibody. AB - Protection against infection with Borrelia burgdorferi is dependent primarily on induction of complement-dependent antibody that can kill the spirochete. Measuring the production of sustained high levels of borreliacidal antibody is thus paramount for determining potential vaccine efficacy. We investigated the borreliacidal antibody response in sera and the amount of antibody produced by cultured lymph node cells of C3H/HeJ mice vaccinated with outer surface protein C (OspC). We showed that recombinant OspC was a weak stimulant of borreliacidal antibody production compared to whole cells of OspC-expressing B. burgdorferi. Mice vaccinated with B. burgdorferi in adjuvant produced a high level (titer, 5,120) of anti-OspC borreliacidal antibody, which waned rapidly. Similarly, borreliacidal antibody production by cultured lymph node cells from vaccinated mice peaked soon after vaccination and then decreased. Treatment of lymph node cells with interleukin-6 (IL-6) augmented borreliacidal antibody production, particularly immunoglobulin G2b, whereas treatment with anti-IL-6 inhibited the borreliacidal response. These findings demonstrate a previously unrecognized role for IL-6 in borreliacidal antibody production that may have important implications for vaccine development. PMID- 11401964 TI - Identification of Legionella pneumophila rcp, a pagP-like gene that confers resistance to cationic antimicrobial peptides and promotes intracellular infection. AB - In the course of characterizing a locus involved in heme utilization, we identified a Legionella pneumophila gene predicted to encode a protein with homology to the product of the Salmonella enterica serovar Typhimurium pagP gene. In Salmonella, pagP increases resistance to the bactericidal effects of cationic antimicrobial peptides (CAMPs). Mutants with insertions in the L. pneumophila pagP-like gene were generated and showed decreased resistance to different structural classes of CAMPs compared to the wild type; hence, this gene was designated rcp for resistance to cationic antimicrobial peptides. Furthermore, Legionella CAMP resistance was induced by growth in low-magnesium medium. To determine whether rcp had any role in intracellular survival, mutants were tested in the two most relevant host cells for Legionnaires' disease, i.e., amoebae and macrophages. These mutants exhibited a 1,000-fold-decreased recovery during a Hartmannella vermiformis coculture. Complementation of the infectivity defect could be achieved by introduction of a plasmid containing the intact rcp gene. Mutations in rcp consistently reduced both the numbers of bacteria recovered during intracellular infection and their cytopathic capacity for U937 macrophages. The rcp mutant was also more defective for lung colonization of A/J mice. Growth of rcp mutants in buffered yeast extract broth was identical to that of the wild type, indicating that the observed differences in numbers of bacteria recovered from host cells were not due to a generalized growth defect. However, in low-Mg(2+) medium, the rcp mutant was impaired in stationary-phase survival. This is the first demonstration of a pagP-like gene, involved in resistance to CAMPs, being required for intracellular infection and virulence. PMID- 11401965 TI - Down-modulation of L-selectin by lipopolysaccharide is not required for lipopolysaccharide-induced expression of CD14 in mouse bone marrow granulocytes. AB - We established in previous studies that a constitutive lipopolysaccharide (LPS) receptor of low affinity is present on mouse bone marrow granulocytes (BMG). This yet-unidentified receptor is involved in the LPS-induced expression of a second LPS receptor, CD14. Because it has been claimed that L-selectin (CD62L) is a low affinity LPS receptor in mature granulocytes (polymorphonuclear leukocytes), it may be asked whether this molecule could be the constitutive LPS receptor in BMG. We show in this study that L-selectin is constitutively present on BMG and is down-regulated after exposure of the cells to LPS. A phorbol ester induced a down regulation of CD62L and blocked the LPS-induced expression of CD14. However, a metalloproteinase inhibitor (BB-3103) blocked the former but not the latter effect of PMA. We also observed an absence of cross-reactivity between LPS and a CD62L ligand (fucoidan) in binding studies with radiolabeled derivatives of the two agents. Furthermore, BMG from L-selectin-deficient mice expressed normal levels of CD14 in response to LPS. Taken together, these results demonstrate that in BMG, L-selectin is not the constitutive LPS receptor required for the LPS induced expression of CD14. PMID- 11401966 TI - Subcellular localization of the Iitracellular survival-enhancing Eis protein of Mycobacterium tuberculosis. AB - Mycobacterium tuberculosis is a facultative intracellular pathogen that has evolved the ability to survive and multiply within human macrophages. It is not clear how M. tuberculosis avoids the destructive action of macrophages, but this ability is fundamental in the pathogenicity of tuberculosis. A gene previously identified in M. tuberculosis, designated eis, was found to enhance intracellular survival of Mycobacterium smegmatis in the human macrophage-like cell line U-937 (J. Wei et al., J. Bacteriol. 182:377-384, 2000). When eis was introduced into M. smegmatis on a multicopy vector, sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the appearance of a unique 42-kDa protein band corresponding to the predicted molecular weight of the eis gene product. This band was electroeluted from the gel with a purity of >90% and subjected to N terminal amino acid sequencing, which demonstrated that the 42-kDa band was indeed the protein product of eis. The Eis protein produced by M. tuberculosis H37Ra had an identical N-terminal amino acid sequence. A synthetic polypeptide corresponding to a carboxyl-terminal region of the deduced eis protein sequence was used to generate affinity-purified rabbit polyclonal antibodies that reacted with the 42-kDa protein in Western blot analysis. Hydropathy profile analysis showed the Eis protein to be predominantly hydrophilic with a potential hydrophobic amino terminus. Phase separation of M. tuberculosis H37Ra lysates by the nonionic detergent Triton X-114 revealed the Eis protein in both the aqueous and detergent phases. After fractionation of M. tuberculosis by differential centrifugation, Eis protein appeared mainly in the cytoplasmic fraction but also in the membrane, cell wall, and culture supernatant fractions as well. Forty percent of the sera from pulmonary tuberculosis patients tested for anti-Eis antibody gave positive reactions in Western blot analysis. Although the function of Eis remains unknown, evidence presented here suggests it associates with the cell surface and is released into the culture medium. It is produced during human tuberculosis infection and therefore may be an important M. tuberculosis immunogen. PMID- 11401967 TI - Impact of genotypic variation of Borrelia burgdorferi sensu stricto on kinetics of dissemination and severity of disease in C3H/HeJ mice. AB - Various genotypes of Borrelia burgdorferi sensu stricto have been previously identified among a large collection of isolates cultured from patients with Lyme disease in the United States. Furthermore, association of specific genotypes with hematogenous dissemination early in the disease course has been observed. The present study assessed kinetics of spirochete dissemination and disease severity in C3H/HeJ mice infected with two different genotypes of B. burgdorferi. Spirochete load in plasma and ear and other tissue samples of infected mice was measured by quantitative PCR, and these data were compared to those obtained by culture and histopathologic analysis. In mice infected with isolate BL206 (a type 1 strain), the peak number of spirochetes was observed in plasma between day 4 and 7, in heart and ear tissue on day 14, and in joints on day 28 postinoculation. There was a correlation between the peak number of spirochetes in plasma on day 4 or 7 and that in ear biopsy and joint specimens on day 14. By contrast, spirochete burdens in plasma of mice infected with isolate B356 (a type 3 strain) were 16- and 5-fold lower than those of BL206-infected mice on days 7 and 14 of infection, respectively. Similarly, approximately 6- and 13-fold fewer spirochetes were detected in the heart tissues of B356-infected mice compared to BL206-infected mice. Histopathologically, severe arthritis and aortitis were noted only in mice infected with isolate BL206. Spirochete dissemination and disease severity vary significantly in mice infected with distinct genotypes of B. burgdorferi, suggesting that genotypic differences in the infecting spirochetes play a key role in the pathogenesis and development of clinical disease. PMID- 11401968 TI - Down-regulated lymphoproliferation coincides with parasite maturation and with the collapse of both gamma interferon and interleukin-4 responses in a bovine model of onchocerciasis. AB - Onchocerciasis is a debilitating parasitic infection caused by the filarial nematode Onchocerca volvulus. Infections are chronic, and persistence of the parasites for several years argues for highly adapted mechanisms of immune evasion. Due to the restricted host repertoire of O. volvulus, we have used the cattle parasite Onchocerca ochengi to investigate the nature of immunomodulation underpinning these long-term infections. Cattle were infected with a single inoculation of 350 infective-stage larvae under laboratory conditions (n = 6). Intradermal nodules containing immature adult worms were detected from 110 days postinfection, and microfilariae in skin were detected from day 280 postinfection. Parasite-specific responses during early infection were nonpolarized with respect to the major Th cytokines (interleukin-4 [IL-4], IL-2, and gamma interferon [IFN-gamma]) produced by antigen-stimulated peripheral blood mononuclear cells (PBMC) or serum antibody isotypes. Antigen-induced proliferation of PBMC peaked shortly after exposure and remained high during the prepatent infection. As the parasites matured and animals developed patent infections, there was a profound down-regulation of lymphoproliferation, accompanied by sharp falls in the expression of both IL-4 and IFN-gamma and a gradual decline in IL-2. Levels of immunoglobulin G2 (IgG2) fell, while those of IgG1 remained high. We conclude that neither a classical Th2 response nor a simple Th1-to-Th2 switch is sufficient to explain the immunomodulation associated with patent Onchocerca infections. Instead, there is an initial Th0 response, which matures into a response with some, but not all of the features of a Th2 response. The natural host-parasite relationship of O. ochengi in cattle may be useful as both a descriptive and predictive tool to test more refined models of immunomodulation in onchocerciasis. PMID- 11401969 TI - CD8(+) T cells participate in the memory immune response to Mycobacterium tuberculosis. AB - The contribution of CD8(+) T cells to the control of tuberculosis has been studied primarily during acute infection in mouse models. Memory or recall responses in tuberculosis are less well characterized, particularly with respect to the CD8 T-cell subset. In fact, there are published reports that CD8(+) T cells do not participate in the memory immune response to Mycobacterium tuberculosis. We examined the CD8(+) T-cell memory and local recall response to M. tuberculosis. To establish a memory immunity model, C57BL/6 mice were infected with M. tuberculosis, followed by treatment with anti-mycobacterial drugs and prolonged rest. The lungs of memory immune mice contained CD4(+) and CD8(+) T cells with the cell surface phenotype characteristic of memory cells (CD69(low) CD25(low) CD44(high)). At 1 week postchallenge with M. tuberculosis via aerosol, > or =30% of both CD4(+) and CD8(+) T cells in the lungs of immune mice expressed the activation marker CD69 and could be restimulated to produce gamma interferon (IFN-gamma). In contrast, <6% of T cells in the lungs of naive challenged mice were CD69(+) at 1 week postchallenge, and IFN-gamma production was not observed at this time point. CD8(+) T cells from the lungs of both naive and memory mice after challenge were cytotoxic toward M. tuberculosis-infected macrophages. Our data indicate that memory and recall immunity to M. tuberculosis is comprised of both CD4(+) and CD8(+) T lymphocytes and that there is a rapid response of both subsets in the lungs following challenge. PMID- 11401971 TI - Primary and booster mucosal immune responses to meningococcal group A and C conjugate and polysaccharide vaccines administered to university students in the United Kingdom. AB - Meningococcal group A+C capsular polysaccharide (PS) conjugate vaccines may prime for serum immunoglobulin G (IgG) memory responses to meningococcal capsular PS. It is not known whether these vaccines induce immunological memory at the mucosal level, which may be important in reducing nasopharyngeal carriage. Mucosal immune responses to meningococcal conjugate and PS vaccines in young adults were investigated. Healthy university students were randomized to receive either a groups A+C meningococcal conjugate vaccine (MACconj, n = 100) or a group A+C meningococcal PS vaccine (MACPS, n = 95). One year after the primary immunization, both groups were randomized again to receive a MACconj or a MACPS booster vaccination. Saliva samples were collected before and 1 month after the primary and booster vaccinations. Anti-meningococcal A (MenA) and C (MenC) PS IgA and IgG antibody levels were measured by a standard enzyme-linked immunosorbent assay. After the primary vaccination, salivary MenA and MenC IgG and MenA IgA concentrations were significantly increased after immunization with both MACconj and MACPS vaccines, but the salivary Men C IgA level was increased only after MACPS vaccine (P < 0.01). IgA responses to both serogroups were greater for MACPS than MACconj vaccine (P < 0.05), whereas no significant differences were seen for IgG responses. MenA IgG titers were higher after the MACPS booster in MACconj primed subjects than after the MACPS primary vaccination, suggesting the presence of IgG memory. Antibody responses to a dose of either MACPS or MACconj were not significantly reduced in those previously given MACPS compared to the primary responses to those vaccines. Meningococcal A+C conjugate and PS vaccines induce significant mucosal responses in young adults. MACconj priming may induce IgG memory at the mucosal level, which is likely to be a reflection of an anamnestic serum IgG response. No evidence of mucosal hyporesponsiveness was observed after MACPS priming in this study. PMID- 11401970 TI - Antigenic diversity among Helicobacter pylori vacuolating toxins. AB - Helicobacter pylori vacuolating cytotoxin (VacA) is a secreted protein that induces vacuolation of epithelial cells. To study VacA structure and function, we immunized mice with purified type s1-m1 VacA from H. pylori strain 60190 and generated a panel of 10 immunoglobulin G1kappa anti-VacA monoclonal antibodies. All of the antibodies reacted with purified native VacA but not with denatured VacA, suggesting that these antibodies react with conformational epitopes. Seven of the antibodies reacted with both native and acid-treated VacA, which suggests that epitopes present on both oligomeric and monomeric forms of the toxin were recognized. Two monoclonal antibodies, both reactive with epitopes formed by amino acids in the carboxy-terminal portion of VacA (amino acids 685 to 821), neutralized the cytotoxic activity of type s1-m1 VacA when toxin and antibody were mixed prior to cell contact but failed to neutralize the cytotoxic activity of type s1-m2 VacA. Only 3 of the 10 antibodies consistently recognized type s1 m1 VacA toxins from multiple H. pylori strains, and none of the antibodies recognized type s2-m2 VacA toxins. These results indicate that there is considerable antigenic diversity among VacA toxins produced by different H. pylori strains. PMID- 11401972 TI - Polysaccharide biosynthesis locus required for virulence of Bacteroides fragilis. AB - Bacteroides fragilis, though only a minor component of the human intestinal commensal flora, is the anaerobe most frequently isolated from intra-abdominal abscesses. B. fragilis 9343 expresses at least three capsular polysaccharides polysaccharide A (PS A), PS B, and PS C. Purified PS A and PS B have been tested in animal models and are both able to induce the formation of intra-abdominal abscesses. Mutants unable to synthesize PS B or PS C still facilitate abscess formation at levels comparable to those of wild-type 9343. To determine the contribution of PS A to abscess formation in the context of the intact organism, the PS A biosynthesis region was cloned, sequenced, and deleted from 9343 to produce a PS A-negative mutant. Animal experiments demonstrate that the abscess inducing capability of 9343 is severely attenuated when the organism cannot synthesize PS A, despite continued synthesis of the other capsular polysaccharides. The PS A of 9343 contains an unusual free amino sugar that is essential for abscess formation by this polymer. PCR analysis of the PS A biosynthesis loci of 50 B. fragilis isolates indicates that regions flanking each side of this locus are conserved in all strains. The downstream conserved region includes two terminal PS A biosynthesis genes that homology-based analyses predict are involved in the synthesis and transfer of the free amino sugar of PS A. Conservation of these genes suggests that this sugar is present in the PS A of all serotypes and may explain the abscessogenic nature of B. fragilis. PMID- 11401973 TI - Dendritic cell activation and cytokine production induced by group B Neisseria meningitidis: interleukin-12 production depends on lipopolysaccharide expression in intact bacteria. AB - Interactions between dendritic cells (DCs) and microbial pathogens are fundamental to the generation of innate and adaptive immune responses. Upon stimulation with bacteria or bacterial components such as lipopolysaccharide (LPS), immature DCs undergo a maturation process that involves expression of costimulatory molecules, HLA molecules, and cytokines and chemokines, thus providing critical signals for lymphocyte development and differentiation. In this study, we investigated the response of in vitro-generated human DCs to a serogroup B strain of Neisseria meningitidis compared to an isogenic mutant lpxA strain totally deficient in LPS and purified LPS from the same strain. We show that the parent strain, lpxA mutant, and meningococcal LPS all induce DC maturation as measured by increased surface expression of costimulatory molecules and HLA class I and II molecules. Both the parent and lpxA strains induced production of tumor necrosis factor alpha (TNF-alpha), interleukin-1alpha (IL 1alpha), and IL-6 in DCs, although the parent was the more potent stimulus. In contrast, high-level IL-12 production was only seen with the parent strain. Compared to intact bacteria, purified LPS was a very poor inducer of IL-1alpha, IL-6, and TNF-alpha production and induced no detectable IL-12. Addition of exogenous LPS to the lpxA strain only partially restored cytokine production and did not restore IL-12 production. These data show that non-LPS components of N. meningitidis induce DC maturation, but that LPS in the context of the intact bacterium is required for high-level cytokine production, especially that of IL 12. These findings may be useful in assessing components of N. meningitidis as potential vaccine candidates. PMID- 11401974 TI - CdtA, CdtB, and CdtC form a tripartite complex that is required for cytolethal distending toxin activity. AB - Campylobacter jejuni encodes a cytolethal distending toxin (CDT) that causes cells to arrest in the G(2)/M transition phase of the cell cycle. Highly related toxins are also produced by other important bacterial pathogens. CDT activity requires the function of three genes: cdtA, cdtB, and cdtC. Recent studies have established that CdtB is the active subunit of CDT, exerting its effect as a nuclease that damages the DNA and triggers cell cycle arrest. Microinjection of CdtB into target cells led to G(2)/M arrest and cytoplasmic distention, in a manner indistinguishable from that caused by CDT treatment. Despite this progress, nothing is known about the composition of the CDT holotoxin or the function of CdtA and CdtC. We show here that, when applied individually, purified CdtA, CdtB, or CdtC does not exhibit toxic activity. In contrast, CdtA, CdtB, and CdtC when combined, interact with one another to form an active tripartite holotoxin that exhibits full cellular toxicity. CdtA has a domain that shares similarity with the B chain of ricin-related toxins. We therefore proposed that CDT is a tripartite toxin composed of CdtB as the enzymatically active subunit and of CdtA and CdtC as the heterodimeric B subunit required for the delivery of CdtB. PMID- 11401975 TI - Role of Enterococcus faecalis surface protein Esp in the pathogenesis of ascending urinary tract infection. AB - Enterococcus faecalis bacteria isolated from patients with bacteremia, endocarditis, and urinary tract infections more frequently express the surface protein Esp than do fecal isolates. To assess the role of Esp in colonization and persistence of E. faecalis in an animal model of ascending urinary tract infection, we compared an Esp(+) strain of E. faecalis to its isogenic Esp deficient mutant. Groups of CBA/J mice were challenged transurethrally with 10(8) CFU of either the parent or mutant strain, and bacteria in the urine, bladder, and kidneys were enumerated 5 days postinfection. Significantly higher numbers of bacteria were recovered from the bladder and urine of mice challenged with the parent strain than from the bladder and urine of mice challenged with the mutant. Colonization of the kidney, however, was not significantly different between the parent and mutant strains. Histopathological evaluations of kidney and bladder tissue done at 5 days postinfection did not show marked histopathological changes consistent with inflammation, mucosal hyperplasia, or apoptosis, and there was no observable difference between the mice challenged with the parent and those challenged with the mutant. We conclude that, while Esp does not influence histopathological changes associated with acute urinary tract infections, it contributes to colonization and persistence of E. faecalis at this site. PMID- 11401976 TI - Establishing a direct role for the Bartonella bacilliformis invasion-associated locus B (IalB) protein in human erythrocyte parasitism. AB - The invasion-associated locus A and B genes (ialAB) of Bartonella bacilliformis were previously shown to confer an erythrocyte-invasive phenotype upon Escherichia coli, indirectly implicating their role in virulence. We report the first direct demonstration of a role for ialB as a virulence factor in B. bacilliformis. The presence of a secretory signal sequence and amino acid sequence similarity to two known outer membrane proteins involved in virulence suggested that IalB was an outer membrane protein. To develop an antiserum for protein localization, the ialB gene was cloned in frame into an expression vector with a six-histidine tag and under control of the lacZ promoter. The IalB fusion protein was purified by nickel affinity chromatography and used to raise polyclonal antibodies. IalB was initially localized to the bacterial membrane fraction. To further localize IalB, B. bacilliformis inner and outer membranes were fractionated by sucrose density gradient centrifugation and identified by appearance, buoyant density (rho), and cytochrome b content. Inner and outer membrane proteins were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and IalB was positively identified by Western blot. Contrary to expectations, IalB was localized to the inner membrane of the pathogen. To directly demonstrate a role for IalB in erythrocyte parasitism, the B. bacilliformis ialB gene was disrupted by insertional mutagenesis. The resulting ialB mutant strain was complemented in trans with a replicative plasmid encoding the full-length ialB gene. PCR and high-stringency DNA hybridization confirmed mutagenesis and transcomplementation events. Abrogation and restoration of ialB expression was verified by SDS-PAGE and immunoblotting. In vitro virulence assays showed that mutagenesis of ialB decreased bacterial association and invasion of human erythrocytes by 47 to 53% relative to controls. Transcomplementation of ialB restored erythrocyte association and invasion rates to levels observed in the parental strain. These data provide direct evidence for IalB's role in erythrocyte parasitism and represent the first demonstration of molecular Koch's postulates for a Bartonella species. PMID- 11401977 TI - Type I Helicobacter pylori lipopolysaccharide stimulates toll-like receptor 4 and activates mitogen oxidase 1 in gastric pit cells. AB - Guinea pig gastric pit cells express an isozyme of gp91-phox, mitogen oxidase 1 (Mox1), and essential components for the phagocyte NADPH oxidase (p67-, p47-, p40 , and p22-phox). Helicobacter pylori lipopolysaccharide (LPS) and Escherichia coli LPS have been shown to function as potent activators for the Mox1 oxidase. These cells spontaneously secreted about 10 nmol of superoxide anion (O(2)(-))/mg of protein/h under LPS-free conditions. They expressed the mRNA and protein of Toll-like receptor 4 (TLR4) but not those of TLR2. LPS from type I H. pylori at 2.1 endotoxin units/ml or higher stimulated TLR4-mediated phosphorylations of transforming growth factor beta-activated kinase 1 and its binding protein 1 induced TLR4 and p67-phox and up-regulated O(2)(-) production 10-fold. In contrast, none of these events occurred with H. pylori LPS from complete or partial deletion mutants of the cag pathogenicity island. Lipid A was confirmed to be a bioactive component for the priming effects, while removal of bisphosphates from lipid A completely eliminated the effects, suggesting the importance of the phosphorylation pattern besides the acylation pattern for the bioactivity. H. pylori LPS is generally accepted as having low toxicity; however, our results suggest that type I H. pylori lipid A may be a potent stimulator for innate immune responses of gastric mucosa by stimulating the TLR4 cascade and Mox1 oxidase in pit cells. PMID- 11401978 TI - Naturally acquired antibody responses to Plasmodium falciparum merozoite surface protein 4 in a population living in an area of endemicity in Vietnam. AB - Merozoite surface protein 4 (MSP4) of Plasmodium falciparum is a glycosylphosphatidylinositol-anchored integral membrane protein that is being developed as a component of a subunit vaccine against malaria. We report here the measurement of naturally acquired antibodies to MSP4 in a population of individuals living in the Khanh-Hoa region of Vietnam, an area where malaria is highly endemic. Antibodies to MSP4 were detected in 94% of the study population at titers of 1:5,000 or greater. Two forms of recombinant MSP4 produced in either Escherichia coli or Saccharomyces cerevisiae were compared as substrates in the enzyme-linked immunosorbent assay. There was an excellent correlation between reactivity measured to either, although the yeast substrate was recognized by a higher percentage of sera. Four different regions of MSP4 were recognized by human antibodies, demonstrating that there are at least four distinct epitopes in this protein. In the carboxyl terminus, where the single epidermal growth factor like domain is located, the reactive epitope(s) was shown to be conformation dependent, as disruption of the disulfide bonds almost completely abolished reactivity with human antibodies. The anti-MSP4 antibodies were mainly of the immunoglobulin G1 (IgG1) and IgG3 subclasses, suggesting that such antibodies may play a role in opsonization and complement-mediated lysis of free merozoites. Individuals in the study population were drug-cured and followed up for 6 months; no significant correlation was observed between the anti-MSP4 antibodies and the absence of parasitemia during the surveillance period. As a comparison, antibodies to MSP1(19), a leading vaccine candidate, were measured, and no correlation with protection was observed in these individuals. The anti-MSP1(19) antibodies were predominantly of the IgG1 isotype, in contrast to the IgG3 predominance noted for MSP4. PMID- 11401979 TI - Growth phase-dependent invasion of Pseudomonas aeruginosa and its survival within HeLa cells. AB - Clinical isolates of Pseudomonas aeruginosa are classified into invasive and noninvasive (cytolytic) strains. In a noninvasive PA103 background, ExoS and ExoT have recently been shown to function as anti-internalization factors. However, these two factors seemed not to have such a function in an invasive strain PAK background. In this study, using HeLa tissue culture cells, we observed that the internalization of invasive strain PAK is dependent on its growth phases, with the stationary-phase cells internalized about 100-fold more efficiently than the exponential-phase cells. This growth phase-dependent internalization was not observed in the noninvasive PA103 strain. Further analysis of various mutant derivatives of the invasive PAK and the noninvasive PA103 strains demonstrated that ExoS or ExoT that is injected into host cells by a type III secretion machinery functions as an anti-internalization factor in both types of strains. In correlation with the growth phase-dependent internalization, the invasive strain PAK translocates much higher amount of ExoS and ExoT into HeLa cells when it is in an exponential-growth phase than when it is in a stationary-growth phase, whereas the translocation of ExoT by the noninvasive strain PA103 is consistently high regardless of the growth phases, suggesting a difference in the regulatory mechanism of type III secretion between the two types of strains. Consistent with the invasive phenotype of the parent strain, an internalized PAK derivative survived well within the HeLa cells, whereas the viability of internalized PA103 derivative was dramatically decreased and completely cleared within 48 h. These results indicate that the invasive strains of P. aeruginosa have evolved the mechanism of intracellular survival, whereas the noninvasive P. aeruginosa strains have lost or not acquired the ability to survive within the epithelial cells. PMID- 11401980 TI - Deletion of wboA enhances activation of the lectin pathway of complement in Brucella abortus and Brucella melitensis. AB - Brucella spp. are gram-negative intracellular pathogens that survive and multiply within phagocytic cells of their hosts. Smooth organisms present O polysaccharides (OPS) on their surface. These OPS help the bacteria avoid the bactericidal action of serum. The wboA gene, coding for the enzyme glycosyltransferase, is essential for the synthesis of O chain in Brucella. In this study, the sensitivity to serum of smooth, virulent Brucella melitensis 16M and B. abortus 2308, rough wboA mutants VTRM1, RA1, and WRR51 derived from these two Brucella species, and the B. abortus vaccine strain RB51 was assayed using normal nonimmune human serum (NHS). The deposition of complement components and mannose-binding lectin (MBL) on the bacterial surface was detected by flow cytometry. Rough B. abortus mutants were more sensitive to the bactericidal action of NHS than were rough B. melitensis mutants. Complement components were deposited on smooth strains at a slower rate compared to rough strains. Deposition of iC3b and C5b-9 and bacterial killing occurred when bacteria were treated with C1q-depleted, but not with C2-depleted serum or NHS in the presence of Mg-EGTA. These results indicate that (i) OPS-deficient strains derived from B. melitensis 16M are more resistant to the bactericidal action of NHS than OPS deficient strains derived from B. abortus 2308, (ii) both the classical and the MBL-mediated pathways are involved in complement deposition and complement mediated killing of Brucella, and (iii) the alternative pathway is not activated by smooth or rough brucellae. PMID- 11401981 TI - Tapeworm infection reduces epithelial ion transport abnormalities in murine dextran sulfate sodium-induced colitis. AB - The rat tapeworm Hymenolepis diminuta was used to test the hypothesis that helminth infection could modulate murine colitis. Mice were infected with five H. diminuta cysticercoids, and colitis was evoked via free access to 4% (wt/vol) dextran sulfate sodium (DSS)-containing drinking water for 5 days. BALB/c mice were either infected with H. diminuta and 7 days later exposed to DSS (prophylactic strategy) or started on DSS and infected with H. diminuta 48 h later (treatment strategy). Naive and H. diminuta-only-infected mice served as controls. On autopsy, colonic segments were processed for histological examination and myeloperoxidase (MPO) measurement or mounted in Ussing chambers for assessment of epithelial ion transport. Cytokines (gamma interferon [IFN gamma], interleukin 12 [IL-12], and IL-10) were measured in serum and colonic tissue homogenates. DSS treatment resulted in reduced ion responses (indicated by short-circuit current [Isc]) to electrical nerve stimulation, the cholinergic agonist carbachol, and the adenylate cyclase activator forskolin compared to controls. H. diminuta infection, either prophylactic or therapeutic, caused a significant (P < 0.05) amelioration of these DSS-induced irregularities in stimulated ion transport. In contrast, the histopathology (i.e., mixed immune cell infiltrate, edema, and ulcerative damage) and elevated MPO levels that accompany DSS colitis were unaffected by concomitant H. diminuta infection. Similarly, there were no significant differences in levels of IFN-gamma, IL-12, or IL-10 in serum or tissue from any of the treatment groups at the time of autopsy. We suggest that abolishment of colitis-induced epithelial ion transport abnormalities by H. diminuta infection provides proof-of-principle data and speculate that helminth therapy may provide relief of disease symptoms in colitis. PMID- 11401982 TI - Activation of interleukin-1 receptor-associated kinase by gram-negative flagellin. AB - Flagellin from various species of gram-negative bacteria activates monocytes to produce proinflammatory cytokines. We have analyzed the pathway by which Salmonella enteritidis flagellin (FliC) activates murine and human monocyte/macrophage-like cell lines. Since lipopolysaccharide (LPS), the principal immune stimulatory component of gram-negative bacteria, is known to signal through Toll-like receptor 4 (TLR4), we tested the possibility that FliC also signals via TLR4. When murine HeNC2 cells were stimulated with LPS in the presence of a neutralizing anti-TLR4 monoclonal antibody, tumor necrosis factor alpha (TNF-alpha) and nitric oxide (NO) production were markedly reduced. In contrast, FliC-mediated TNF-alpha and NO production were minimally affected by the anti-TLR4 antibody. Furthermore, FliC, unlike LPS, stimulated TNF-alpha production in the TLR4 mutant cell line, GG2EE, indicating that TLR4 is not essential for FliC-mediated signaling. To test the possibility that FliC signals via another TLR, we measured FliC-mediated activation of interleukin-1 (IL-1) receptor-associated kinase (IRAK), a central component in IL-1R/TLR signaling. FliC induced IRAK activation in HeNC2 and GG2EE cells as well as in the human promonocytic cell line THP-1. IRAK activation was rapid in HeNC2 cells, with maximal activity observed after 5 min of treatment with FliC. In addition, FliC mediated IRAK activation exhibited the same concentration dependence as was demonstrated for the induction of TNF-alpha. These results represent the first demonstration of IRAK activation by a purified bacterial protein and strongly suggest that a TLR distinct from TLR4 is involved in the macrophage inflammatory response to FliC. PMID- 11401983 TI - Cloning, expression, and characterization of a neuraminidase gene from Arcanobacterium pyogenes. AB - Arcanobacterium pyogenes is an opportunistic pathogen, associated with suppurative infections in domestic animals. In addition to pyolysin, a pore forming, cholesterol-binding toxin, A. pyogenes expresses a number of putative virulence factors, including several proteases and neuraminidase activity. A 3,009-bp gene, nanH, was cloned and sequenced and conferred neuraminidase activity on an Escherichia coli host strain. The predicted 107-kDa NanH protein displayed similarity to a number of bacterial neuraminidases and contained the RIP/RLP motif and five copies of the Asp box motif found in all bacterial neuraminidases. Recombinant His-tagged NanH was found to have pH and temperature optima of 5.5 to 6.0 and 55 degrees C, respectively. Insertional deletion of the nanH gene resulted in the reduction, but not absence, of neuraminidase activity, indicating the presence of a second neuraminidase gene in A. pyogenes. NanH was localized to the A. pyogenes cell wall. A. pyogenes adhered to HeLa, CHO, and MDBK cells in a washing-resistant manner. However, the nanH mutant was not defective for adherence to epithelial cells. The role of NanH in host epithelial cell adherence may be masked by the presence of a second neuraminidase in A. pyogenes. PMID- 11401984 TI - Cloning, overexpression, purification, and immunobiology of an 85-kilodalton outer membrane protein from Haemophilus ducreyi. AB - We have identified an 85-kDa outer membrane protein that is expressed by all tested strains of Haemophilus ducreyi. Studies of related proteins from other pathogenic bacteria, including Haemophilus influenzae, Pasteurella multocida, Neisseria gonorrhoeae, Neisseria meningitidis, and Shigella dysenteriae, suggested a role for these proteins in pathogenesis and immunity. In keeping with the first such described protein from Haemophilus influenzae type B, we termed the H. ducreyi protein D15. The gene encoding the H. ducreyi D15 protein was cloned and sequenced, and the deduced amino acid sequence was found to be most similar to sequences of the D15-related proteins from other Pasteurella spp. The arrangement of the flanking genes was similar to that of H. influenzae Rd and suggested that D15 was part of a multigene operon. Attempts to make a null mutation of the D15 gene were unsuccessful, paralleling results in other D15 gene studies. Overexpression of H. ducreyi D15 in Escherichia coli resulted in a source of recombinant D15 (rD15) from which it was readily purified. rD15 was immunogenic, and it was found that immunization of rabbits with an rD15 vaccine preparation conferred partial protection against a virulent challenge infection. Antisera to an N-terminal peptide recognized all tested strains of H. ducreyi. PMID- 11401985 TI - Novel type of fimbriae encoded by the large plasmid of sorbitol-fermenting enterohemorrhagic Escherichia coli O157:H(-). AB - Sorbitol-fermenting (SF) enterohemorrhagic Escherichia coli (EHEC) O157:H(-) have emerged as important causes of diarrheal diseases and the hemolytic-uremic syndrome in Germany. In this study, we characterized a 32-kb fragment of the plasmid of SF EHEC O157:H(-), pSFO157, which differs markedly from plasmid pO157 of classical non-sorbitol-fermenting EHEC O157:H7. We found a cluster of six genes, termed sfpA, sfpH, sfpC, sfpD, sfpJ, and sfpG, which mediate mannose resistant hemagglutination and the expression of fimbriae. sfp genes are similar to the pap genes, encoding P-fimbriae of uropathogenic E. coli, but the sfp cluster lacks homologues of genes encoding subunits of a tip fibrillum as well as regulatory genes. The major pilin, SfpA, despite its similarity to PapA, does not cluster together with known PapA alleles in a phylogenetic tree but is structurally related to the PmpA pilin of Proteus mirabilis. The putative adhesin gene sfpG, responsible for the hemagglutination phenotype, shows significant homology neither to papG nor to other known sequences. Sfp fimbriae are 3 to 5 nm in diameter, in contrast to P-fimbriae, which are 7 nm in diameter. PCR analyses showed that the sfp gene cluster is a characteristic of SF EHEC O157:H(-) strains and is not present in other EHEC isolates, diarrheagenic E. coli, or other Enterobacteriaceae. The sfp gene cluster is flanked by two blocks of insertion sequences and an origin of plasmid replication, indicating that horizontal gene transfer may have contributed to the presence of Sfp fimbriae in SF EHEC O157:H( ). PMID- 11401986 TI - Analysis of the capsule biosynthetic locus of Mannheimia (Pasteurella) haemolytica A1 and proposal of a nomenclature system. AB - A 16-kbp DNA region that contains genes involved in the biosynthesis of the capsule of Mannheimia (Pasteurella) haemolytica A1 has been characterized. The gene cluster can be divided into three regions like those of the typical group II capsule biosynthetic clusters in gram-negative bacteria. Region 1 contains four genes (wzt, wzm, wzf, and wza) which code for an ATP-binding cassette transport apparatus for the secretion of the capsule materials across the membranes. The M. haemolytica A1 wzt and wzm genes were able to complement Escherichia coli kpsT and kpsM mutants, respectively. Further, the ATP binding activity of Wzt was demonstrated by its affinity for ATP-agarose, and the lipoprotein nature of Wza was supported by [(3)H]palmitate labeling. Region 2 contains six genes; four genes (orf1/2/3/4) code for unique functions for which no homologues have been identified to date. The remaining two genes (nmaA and nmaB) code for homologues of UDP-N-acetylglucosamine-2-epimerase and UDP-N-acetylmannosamine dehydrogenase, respectively. These two proteins are highly homologous to the E. coli WecB and WecC proteins (formerly known as RffE and RffD), which are involved in the biosynthesis of enterobacterial common antigen (ECA). Complementation of an E. coli rffE/D mutant with the M. haemolytica A1 nmaA/B genes resulted in the restoration of ECA biosynthesis. Region 3 contains two genes (wbrA and wbrB) which are suggested to be involved in the phospholipid modification of capsular materials. PMID- 11401987 TI - Cloning and expression of two novel hemin binding protein genes from Treponema denticola. AB - Treponema denticola does not appear to produce siderophores, so it must acquire iron by other pathways. Indeed, T. denticola has been shown to have an iron regulated 44-kDa outer membrane protein (HbpA) with hemin binding ability. To characterize the HbpA protein, its gene was cloned from genomic DNA libraries of T. denticola. Sequence analysis of the hbpA open reading frame indicated that it encoded a 42.8-kDa protein with a 23-amino-acid signal peptide. HbpA has no significant homology to any proteins in the databases. Southern blot analysis demonstrated that hbpA is present in several T. denticola ATCC strains and clinical isolates, but not in Treponema pectinovorum, Treponema socranskii, or Escherichia coli. HbpA, expressed as a recombinant protein in E. coli and purified by antibody affinity chromatography, has hemin binding activity as determined by lithium dodecyl sulfate-polyacrylamide gel electrophoresis with tetramethylbenzidine staining. Northern blot analysis showed that there were two hbpA-containing transcripts, of approximately 1.3 and 2.6 kb, and that the RNA levels were low-iron induced. Interestingly, the 2.6-kb mRNA also encoded a second protein with significant homology to hbpA. This downstream gene, called hbpB, was cloned and sequenced and its product was expressed as a fusion protein in E. coli. The hbpB gene product is 49% identical to HbpA and binds hemin. Thus, T. denticola has two novel hemin binding proteins which may be part of a previously unrecognized iron acquisition pathway. PMID- 11401988 TI - Adherence properties of Staphylococcus aureus under static and flow conditions: roles of agr and sar loci, platelets, and plasma ligands. AB - Global regulatory genes in Staphylococcus aureus, including agr and sar, are known to regulate the expression of multiple virulence factors, including cell wall adhesins. In the present study, the adherence of S. aureus RN6390 (wild type), RN6911 (agr), ALC136 (sar), and ALC135 (agr sar) to immobilized fibrinogen, fibronectin, von Willebrand factor (vWF), extracellular matrix (ECM), and human endothelial cells (EC) EAhy.926 was studied. Bacteria grown to postexponential phase were subjected to light oscillation (static condition) or to shear stress at 200 s(-1) (flow condition) on tissue culture polystyrene plates coated with either protein ligands, ECM, or EC. Adherence of nonlabeled bacteria to immobilized ligands was measured by an image analysis system, while adherence of [(3)H]thymidine-labeled S. aureus to ECM and EC was measured by a beta-scintillation counter. The results showed increased adherence of agr and agr sar mutants to immobilized fibrinogen and higher potential of these mutants to induce platelet aggregation in suspension, decreased adherence of sar and agr sar mutants to immobilized fibronectin and vWF as well as to ECM and EC, increased adherence of both S. aureus wild type and sar mutant to EC treated with platelet rich plasma (PRP) compared to platelet-poor plasma (PPP) and to EC treated with PPP compared to the control, and increased adherence of S. aureus wild type to EC coated with PRP in which platelets were activated with phorbol 12-myristate 13 acetate compared to intact PRP. This finding paralleled the increased adherence to EC of activated compared to intact platelets. It is suggested that platelet mediated S. aureus adherence to EC depends on platelet activation and the number of adherent platelets and available receptors on the platelet membrane. In conclusion, the agr locus downregulates S. aureus adherence to fibrinogen, while the sar locus upregulates S. aureus adherence to fibronectin, vWF, ECM, and EC. The effect of both agr and sar on S. aureus adherence properties develops primarily under flow conditions, which suggests different adhesion mechanisms in static and flow conditions. PMID- 11401989 TI - Evidence of recombination in Porphyromonas gingivalis and random distribution of putative virulence markers. AB - The association of Porphyromonas gingivalis to periodontal disease is not clearly understood. Similar proportions of P. gingivalis may be cultivated from both inactive and actively degrading periodontal pockets. Differences in virulence among strains of P. gingivalis exist, but the molecular reason for this remains unknown. We examined the population structure of P. gingivalis to obtain a framework in which to study pathogenicity in relation to evolution. Phylogenetic trees derived from the sequencing of fragments of four housekeeping genes, ahp, thy, rmlB, and infB, in 57 strains were completely different with no correlation between clustering of strains in the four dendrograms. Combining the various alleles of the four gene fragments sequenced resulted in 41 different sequence types. The index of association, I(A), based on a single representative of each sequence type was 0.143 +/- 0.202, indicating a population at linkage equilibrium. Inclusion of all isolates for the calculation of I(A) resulted in a value of 0.206 +/- 0.171. This suggests an epidemic population structure supported by the finding of genetically identical strains in different parts of the world. We observed a random distribution of two virulence-associated mobile genetic elements, the ragB locus and the insertion sequence IS1598, among 132 strains tested. In conclusion, P. gingivalis has a nonclonal population structure characterized by frequent recombination. Our study suggests that particular genotypes, possibly with increased pathogenic potential, may spread successfully in the human population. PMID- 11401990 TI - Distinct cytokine regulation by cholera toxin and type II heat-labile toxins involves differential regulation of CD40 ligand on CD4(+) T cells. AB - Cholera toxin (CT) and the type II heat-labile enterotoxins (HLT) LT-IIa and LT IIb act as potent systemic and mucosal adjuvants and induce distinct T-helper (Th)-cell cytokine profiles. In the present study, CT and the type II HLT were found to differentially affect cytokine production by anti-CD3-stimulated human peripheral blood mononuclear cells (PBMC), and the cellular mechanisms responsible were investigated. CT suppressed interleukin-2 (IL-2), tumor necrosis factor alpha (TNF-alpha), and IL-12 production by PBMC cultures more than either LT-IIa or LT-IIb. CT but not LT-IIa or LT-IIb reduced the expression of CD4(+) T cell surface activation markers (CD25 and CD69) and subsequent proliferative responses of anti-CD3-stimulated T cells. CT but not LT-IIa or LT-IIb significantly reduced the expression of CD40 ligand (CD40L) on CD4(+) T cells. In a coculture system, CT-treated CD4(+) T cells induced significantly less TNF alpha and IL-12 p70 production by both autologous monocytes and monocyte-derived dendritic cells than either LT-IIa- or LT-IIb-treated CD4(+) T cells. These findings demonstrate that CT, LT-IIa, and LT-IIb differentially affect CD40-CD40L interactions between antigen-presenting cells and T cells and help explain the distinct cytokine profiles observed with type I and type II HLT when used as mucosal adjuvants. PMID- 11401991 TI - Porphyromonas gingivalis gingipain-R enhances interleukin-8 but decreases gamma interferon-inducible protein 10 production by human gingival fibroblasts in response to T-cell contact. AB - Proteases produced by Porphyromonas gingivalis, an oral pathogen, are considered important virulence factors and may affect the responses of cells equipped with proteinase-activated receptors. The aim of this study was to investigate the effect of the arginine-specific cysteine protease gingipain-R produced by P. gingivalis on chemokine production by human gingival fibroblasts (HGF) and the effect of gingipain-R treatment on the subsequent contact-dependent activation of HGF by T cells. HGF incubated in the presence of purified 47-kDa gingipain-R showed increased levels of interleukin-8 (IL-8) mRNA. Cyclooxygenase-2 (COX-2) mRNA was also induced. Further exposure of HGF to activated T cells resulted in the dose- and time-dependent enhancement of IL-8 transcription and release. T cell membrane-bound tumor necrosis factor (TNF) was the ligand inducing IL-8 production by HGF, since TNF neutralization abrogated HGF responses to T-cell contact. The enhanced IL-8 release was due, at least in part, to prostaglandin E(2) production, which was mostly blocked by indomethacin. Gingipain-R proteolytic activity was required since heat inactivation, specific synthetic protease inhibitors, and the natural substrate competitor histatin 5 abrogated its effects. The enhanced production of IL-8 in response to T-cell contact was specific since monocyte chemotactic protein-1 (MCP-1) production was unaffected while interferon-gamma-inducible protein-10 (IP-10) was inhibited. The sum of these activities may result in the recruitment of differential cell types to sites of inflammation since IL-8 preferentially recruits neutrophils and IP-10 attracts activated T cells and may be relevant to the pathogenesis of periodontitis. PMID- 11401992 TI - Immune response induction and new effector mechanisms possibly involved in protection conferred by the Cuban anti-meningococcal BC vaccine. AB - This report explores the participation of some afferent mechanisms in the immune response induced by the Cuban anti-meningococcal vaccine VA-MENGOC-BC. The induction of delayed-type hypersensitivity in nursing babies and lymphocyte proliferation after immunization is demonstrated. The presence of gamma interferon IFN-gamma and interleukin-2 (IL-2) mRNAs but absence of IL-4, IL-5, and IL-10 mRNAs were observed in peripheral blood mononuclear cells from immunized subjects after in vitro challenge with outer membrane vesicles. In addition, some effector functions were also explored. The presence of opsonic activity was demonstrated in sera from vaccinees. The role of neutrophils as essential effector cells was shown. In conclusion, we have shown that, at least in the Cuban adult population, VA-MENGOC-BC induces mechanisms with a T-helper 1 pattern in the afferent and effector branches of the immune response. PMID- 11401993 TI - Protection against anthrax lethal toxin challenge by genetic immunization with a plasmid encoding the lethal factor protein. AB - The ability of genetic vaccination to protect against a lethal challenge of anthrax toxin was evaluated. BALB/c mice were immunized via gene gun inoculation with eucaryotic expression vector plasmids encoding either a fragment of the protective antigen (PA) or a fragment of lethal factor (LF). Plasmid pCLF4 contains the N-terminal region (amino acids [aa] 10 to 254) of Bacillus anthracis LF cloned into the pCI expression plasmid. Plasmid pCPA contains a biologically active portion (aa 175 to 764) of B. anthracis PA cloned into the pCI expression vector. One-micrometer-diameter gold particles were coated with plasmid pCLF4 or pCPA or a 1:1 mixture of both and injected into mice via gene gun (1 microg of plasmid DNA/injection) three times at 2-week intervals. Sera were collected and analyzed for antibody titer as well as antibody isotype. Significantly, titers of antibody to both PA and LF from mice immunized with the combination of pCPA and pCLF4 were four to five times greater than titers from mice immunized with either gene alone. Two weeks following the third and final plasmid DNA boost, all mice were challenged with 5 50% lethal doses of lethal toxin (PA plus LF) injected intravenously into the tail vein. All mice immunized with pCLF4, pCPA, or the combination of both survived the challenge, whereas all unimmunized mice did not survive. These results demonstrate that DNA-based immunization alone can provide protection against a lethal toxin challenge and that DNA immunization against the LF antigen alone provides complete protection. PMID- 11401994 TI - Long-term pertussis-specific immunity after primary vaccination with a combined diphtheria, tetanus, tricomponent acellular pertussis, and hepatitis B vaccine in comparison with that after natural infection. AB - The aim of this study was to compare pertussis-specific humoral and cellular immunity in children 5 years after a primary vaccination with a combined diphtheria, tetanus, tricomponent acellular pertussis, and hepatitis B vaccine (DTaP-HBV; InfanrixHepB; SmithKline Beecham) with immunity after natural infection. The subjects were 38 children aged 5 to 6 years who received DTaP-HBV at 3, 5, and 11 months of life and 21 subjects of similar ages and sex who acquired pertussis in the first year of life. Immunoglobulin G (IgG) antibody titers against Bordetella pertussis antigens, peripheral blood mononuclear cell specific proliferation, and the secretion of cytokines were evaluated. After 5 years, only a small proportion of vaccinated and infected children had significant specific concentrations of IgG in serum against all three B. pertussis antigens, and T-cell responses persisted in a minority of subjects. A preferential type 1 cytokine response with the secretion of gamma interferon was observed in the pertussis group, whereas a type 2 skewed response was observed in the vaccinated children; however, the quantitative differences in the cytokines produced by DTaP-HBV and natural infection were minimal. In conclusion, our results show that the immune responses induced by primary pertussis vaccination are qualitatively and quantitatively similar to those seen in children who recovered from natural infection and highlight the need for booster immunization with pertussis vaccines in order to maintain adequate levels of a specific immune response to B. pertussis. PMID- 11401995 TI - Protection against pulmonary infection with Pseudomonas aeruginosa following immunization with P. aeruginosa-pulsed dendritic cells. AB - To develop a Pseudomonas aeruginosa vaccine that allows the host immune system to select the antigens, we hypothesized that dendritic cells (DC) pulsed with P. aeruginosa would induce protective immunity against pulmonary infections with P. aeruginosa. Incubation of murine bone marrow-derived DC with P. aeruginosa in vitro led to uptake of P. aeruginosa and activation of the DC. Spleen-derived CD4(+) cells from mice immunized with P. aeruginosa-pulsed DC showed increased proliferation, demonstrating that DC pulsed with P. aeruginosa were capable of eliciting a P. aeruginosa-specific immune response. To evaluate if P. aeruginosa pulsed DC can induce protective immunity against P. aeruginosa pulmonary infection, DC incubated with P. aeruginosa in vitro were administered systemically to syngeneic mice, and the mice were then challenged by intrapulmonary infection with P. aeruginosa (5 x 10(4) CFU/mouse) 13 days later. Unimmunized control mice and mice who had previously received naive DC or DC stimulated with lipopolysaccharide or Escherichia coli died within 72 h. In contrast, 45% of mice receiving P. aeruginosa-pulsed DC demonstrated prolonged survival (>14 days). Finally, DC-pulsed with heat-inactivated P. aeruginosa protected CD8(-/-) but not CD4(-/-) mice, demonstrating that CD4(+) T cells were required for the DC pulsed with P. aeruginosa to induce protective immunity. PMID- 11401996 TI - Brucella abortus cyclic beta-1,2-glucan mutants have reduced virulence in mice and are defective in intracellular replication in HeLa cells. AB - Null cyclic beta-1,2-glucan synthetase mutants (cgs mutants) were obtained from Brucella abortus virulent strain 2308 and from B. abortus attenuated vaccinal strain S19. Both mutants show greater sensitivity to surfactants like deoxycholic acid, sodium dodecyl sulfate, and Zwittergent than the parental strains, suggesting cell surface alterations. Although not to the same extent, both mutants display reduced virulence in mice and defective intracellular multiplication in HeLa cells. The B. abortus S19 cgs mutant was completely cleared from the spleens of mice after 4 weeks, while the 2308 mutant showed a 1.5-log reduction of the number of brucellae isolated from the spleens after 12 weeks. These results suggest that cyclic beta-1,2-glucan plays an important role in the residual virulence of the attenuated B. abortus S19 strain. Although the cgs mutant was cleared from the spleens earlier than the wild-type parental strain (B. abortus S19) and produced less inflammatory response, its ability to confer protection against the virulent strain B. abortus 2308 was fully retained. Equivalent levels of induction of spleen gamma interferon mRNA and anti lipopolysaccharide (LPS) of immunoglobulin G2a (IgG2a) subtype antibodies were observed in mice injected with B. abortus S19 or the cgs mutant. However, the titer of anti-LPS antibodies of the IgG1 subtype induced by the cgs mutant was lower than that observed with the parental S19 strain, thus suggesting that the cgs mutant induces a relatively exclusive Th1 response. PMID- 11401997 TI - Traversal of Candida albicans across human blood-brain barrier in vitro. AB - Candida albicans is an opportunistic pathogen, which primarily affects neonates and immunocompromised individuals. The pathogen can invade the central nervous system, resulting in meningitis. At present, the pathogenesis of C. albicans meningitis is unclear. We used an in vitro model of the human blood-brain barrier to investigate the interaction(s) of C. albicans with human brain microvascular endothelial cells (BMEC). Binding of C. albicans to human BMEC was time and inoculum dependent. Invasion of C. albicans into human BMEC was demonstrated by using an enzyme-linked immunosorbent assay based on fluorescent staining of C. albicans with calcoflour. In contrast, avirulent Candida mutant strains and nonpathogenic yeast Saccharomyces cerevisiae were not able to bind and invade human BMEC. Morphological studies revealed that on association with human BMEC, C. albicans formed germ tubes and was able to bud intracellularly. Transmission electron microscopy showed various stages of C. albicans interactions with human BMEC, e.g., pseudopod-like structures on human BMEC membrane and intracellular vacuole-like structures retaining C. albicans. Of interest, C. albicans was able to bud and develop pseudohyphae inside human BMEC without apparent morphological changes of the host cells. In addition, C. albicans penetrates through human BMEC monolayers without a detectable change in transendothelial electrical resistance and inulin permeability. This is the first demonstration that C. albicans is able to adhere, invade, and transcytose across human BMEC without affecting monolayer integrity. A complete understanding of the interaction(s) of C. albicans with human BMEC should contribute to the understanding of the pathogenic mechanism(s) of C. albicans meningitis. PMID- 11401998 TI - Safety and immunogenicity of a proteosome-Shigella flexneri 2a lipopolysaccharide vaccine administered intranasally to healthy adults. AB - We studied the safety and immunogenicity of a Shigella flexneri 2a vaccine comprising native S. flexneri 2a lipopolysaccharide (LPS) complexed to meningococcal outer membrane proteins-proteosomes-in normal, healthy adults. A two-dose series of immunizations was given by intranasal spray, and doses of 0.1, 0.4, 1.0, and 1.5 mg (based on protein) were studied in a dose-escalating design. The vaccine was generally well tolerated. The most common reactions included rhinorrhea and nasal stuffiness, which were clearly dose related (P < or = 0.05). These reactions were self-limited and generally mild. The vaccine elicited S. flexneri 2a LPS-specific immunoglobulin A (IgA), IgG, and IgM antibody-secreting cells (ASCs) in a dose-responsive manner. At doses of 1.0 or 1.5 mg, highly significant (P < 0.001) increases in ASCs of all antibody isotypes occurred and 95% of subjects had an ASC response in at least one antibody isotype. Dose related serum antibody responses were observed, with geometric mean two- to fivefold rises in specific serum IgA and IgG titers and two- to threefold rises in IgM in the 1.0- and 1.5-mg-dose groups (P < 0.0001 for each isotype). Elevated serum antibody levels persisted through day 70. Increases in fecal IgG and IgA and also in urinary IgA specific for S. flexneri 2a LPS were demonstrated. These were most consistent and approached statistical significance (P = 0.02 to 0.12 for various measures) on day 70 after the first dose. The magnitude of immune responses to intranasally administered proteosome-S. flexneri 2a LPS vaccine is similar to those reported for live vaccine candidates associated with protective efficacy in human challenge models, and further evaluation of this product is warranted. PMID- 11401999 TI - Brain granulomas in neurocysticercosis patients are associated with a Th1 and Th2 profile. AB - Neurocysticercosis (NCC) is a common central nervous system (CNS) infection caused by Taenia solium metacestodes. Despite the well-documented importance of the granulomatous response in the pathogenesis of this infection, there is limited information about the types of cells and cytokines involved. In fact, there has been limited characterization of human brain granulomas with any infectious agent. In the present study a detailed histological and immunohistochemical analysis of the immune response was performed on eight craniotomy specimens where a granuloma surrounded each T. solium metacestode. The results indicated that in all the specimens there was a dying parasite surrounded by a mature granuloma with associated fibrosis, angiogenesis, and an inflammatory infiltrate. The most abundant cell types were plasma cells, B and T lymphocytes, macrophages, and mast cells. Th1 cytokines were prevalent and included gamma interferon, interleukin-18 (IL-18), and the immunosuppressive, fibrosis-promoting cytokine transforming growth factor beta. The Th2 cytokines IL-4, IL-13, and IL 10 were also present. These observations indicate that a chronic immune response is elicited in the CNS environment with multiple cell types that together secrete inflammatory and anti-inflammatory cytokines. In addition, both collagen type I and type III deposits were evident and could contribute to irreversible nervous tissue damage in NCC patients. PMID- 11402000 TI - Endogenous interleukin-10 is required for prevention of a hyperinflammatory intracerebral immune response in Listeria monocytogenes meningoencephalitis. AB - To analyze the role of interleukin-10 (IL-10) in bacterial cerebral infections, we studied cerebral listeriosis in IL-10-deficient (IL-10(-/-)) and wild-type (WT) mice, the latter of which express high levels of IL-10 in both primary and secondary cerebral listeriosis. IL-10(-/-) mice succumbed to primary as well as secondary listeriosis, whereas WT mice were significantly protected from secondary listeriosis by prior intraperitoneal immunization with Listeria monocytogenes. Meningoencephalitis developed in both strains; however, in IL-10( /-) mice the inflammation was more severe and associated with increased brain edema and multiple intracerebral hemorrhages. IL-10(-/-) mice recruited significantly increased numbers of leukocytes, in particular granulocytes, to the brain, and the intracerebral cytokine (tumor necrosis factor, IL-1, IL-12, gamma interferon, and inducible nitric oxide synthase) and chemokine (crg2/IP-10, RANTES, MuMig, macrophage inflammatory protein 1alpha [MIP-1alpha], and MIP 1beta) transcription was enhanced compared to that in WT mice. Despite this prominent hyperinflammation, the frequencies of intracerebral L. monocytogenes specific CD8(+) T cells were reduced and the intracerebral bacterial load was not reduced in IL-10(-/-) mice compared to WT mice. Following intraperitoneal infection, IL-10(-/-) mice exhibited hepatic hyperinflammation without better bacterial clearance; however, in contrast to the mice with cerebral listeriosis, they did not succumb, illustrating that intrinsic factors of the target organ have a strong impact on the course and outcome of the infection. PMID- 11402001 TI - Establishment of a persistent Escherichia coli reservoir during the acute phase of a bladder infection. AB - The vast majority of urinary tract infections are caused by strains of uropathogenic Escherichia coli that encode filamentous adhesive organelles called type 1 pili. These structures mediate both bacterial attachment to and invasion of bladder epithelial cells. However, the mechanism by which type 1 pilus mediated bacterial invasion contributes to the pathogenesis of a urinary tract infection is unknown. Here we show that type 1-piliated uropathogens can invade the superficial epithelial cells that line the lumenal surface of the bladder and subsequently replicate, forming massive foci of intracellular E. coli termed bacterial factories. In response to infection, superficial bladder cells exfoliate and are removed with the flow of urine. To avoid clearance by exfoliation, intracellular uropathogens can reemerge and eventually establish a persistent, quiescent bacterial reservoir within the bladder mucosa that may serve as a source for recurrent acute infections. These observations suggest that urinary tract infections are more chronic and invasive than generally assumed. PMID- 11402002 TI - Decreased apoptosis in the ileum and ileal Peyer's patches: a feature after infection with rabbit enteropathogenic Escherichia coli O103. AB - Significant changes occur in intestinal epithelial cells after infection with enteropathogenic Escherichia coli (EPEC). However, it is unclear whether this pathogen alters rates of apoptosis. By using a naturally occurring weaned rabbit infection model, we determined physiological levels of apoptosis in rabbit ileum and ileal Peyer's patches (PP) and compared them to those found after infection with adherent rabbit EPEC (REPEC O103). Various REPEC O103 strains were first tested in vitro for characteristic virulence features. Rabbits were then inoculated with the REPEC O103 strains that infected cultured cells the most efficiently. After experimental infection, intestinal samples were examined by light and electron microscopy. Simultaneously, ileal apoptosis was assessed by using terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) and caspase 3 assays and by apoptotic cell counts based on morphology (hematoxylin-and-eosin staining). The highest physiological apoptotic indices were measured in PP germinal centers (median = 14.7%), followed by PP domed villi (8.1%), tips of absorptive villi (3.8%), and ileal crypt regions (0.5%). Severe infection with REPEC O103 resulted in a significant decrease in apoptosis in PP germinal centers (determined by TUNEL assay; P = 0.01), in the tips of ileal absorptive villi (determined by H&E staining; P = 0.04), and in whole ileal cell lysates (determined by caspase 3 assay; P = 0.001). We concluded that REPEC O103 does not promote apoptosis. Furthermore, we cannot rule out the possibility that REPEC O103, in fact, decreases apoptotic levels in the rabbit ileum. PMID- 11402003 TI - Gram-positive and gram-negative bacteria do not trigger monocytic cytokine production through similar intracellular pathways. AB - Toll-like receptors (TLRs) are involved in human monocyte activation by lipopolysaccharide (LPS) and Staphylococcus aureus Cowan (SAC), suggesting that gram-positive and gram-negative bacteria may trigger similar intracellular events. Treatment with specific kinase inhibitors prior to cell stimulation dramatically decreased LPS-induced cytokine production. Blocking of the p38 pathway prior to LPS stimulation decreased interleukin-1alpha (IL-1alpha), IL 1ra, and tumor necrosis factor alpha (TNF-alpha) production, whereas blocking of the ERK1/2 pathways inhibited IL-1alpha, IL-1beta, and IL-1ra but not TNF-alpha production. When cells were stimulated by SAC, inhibition of the p38 pathway did not affect cytokine production, whereas only IL-1alpha production was decreased in the presence of ERK kinase inhibitor. We also demonstrated that although LPS and SAC have been shown to bind to CD14 before transmitting signals to TLR4 and TLR2, respectively, internalization of CD14 occurred only in monocytes triggered by LPS. Pretreatment of the cells with SB203580, U0126, or a mixture of both inhibitors did not affect internalization of CD14. Altogether, these results suggest that TLR2 signaling does not involve p38 mitogen-activated protein kinase signaling pathways, indicating that divergent pathways are triggered by gram positive and gram-negative bacteria, thereby inducing cytokine production. PMID- 11402004 TI - Homogeneity of antibody responses in tuberculosis patients. AB - The goals of the present study were twofold: (i) to compare the repertoires of antigens in culture filtrates of in vitro-grown Mycobacterium tuberculosis that are recognized by antibodies from noncavitary and cavitary tuberculosis (TB) patients and (ii) to determine the extent of variation that exists between the antigen profiles recognized by individual TB patients. Lipoarabinomannan-free culture filtrate proteins of M. tuberculosis were fractionated by one-dimensional (1-D) and 2-D polyacrylamide gel electrophoresis, and the Western blots were probed with sera from non-human immunodeficiency virus (non-HIV)-infected cavitary and noncavitary TB patients and from HIV-infected, noncavitary TB patients. In contrast to earlier studies based on recombinant antigens of M. tuberculosis which suggested that antibody responses in TB patients were heterogeneous (K. Lyashchenko et al., 1998, Infect. Immun. 66:3936-3940, 1998), our studies with native culture filtrate proteins show that the antibody responses in TB patients show significant homogeneity in being directed against a well-defined subset of antigens. Thus, there is a well-defined subset of culture filtrate antigens that elicits antibodies during noncavitary and cavitary disease. In addition, another set of antigens is recognized primarily by cavitary TB patients. The mapping with individual patient sera presented here suggests that serodiagnostic tests based on the subset of antigens recognized during both noncavitary and cavitary TB will enhance the sensitivity of antibody detection in TB patients, especially in difficult-to-diagnose, smear-negative, noncavitary TB patients. PMID- 11402005 TI - Salmonella-induced cell death is not required for enteritis in calves. AB - Salmonella enterica serovar Typhimurium causes cell death in bovine monocyte derived and murine macrophages in vitro by a sipB-dependent mechanism. During this process, SipB binds and activates caspase-1, which in turn activates the proinflammatory cytokine interleukin-1beta through cleavage. We used bovine ileal ligated loops to address the role of serovar Typhimurium-induced cell death in induction of fluid accumulation and inflammation in this diarrhea model. Twelve perinatal calves had 6- to 9-cm loops prepared in the terminal ileum. They were divided into three groups: one group received an intralumen injection of Luria Bertani broth as a control in 12 loops. The other two groups (four calves each) were inoculated with 0.75 x 10(9) CFU of either wild-type serovar Typhimurium (strain IR715) or a sopB mutant per loop in 12 loops. Hematoxylin and eosin stained sections were scored for inflammation, and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL) positive cells were detected in situ. Fluid accumulation began at 3 h postinfection (PI). Inflammation was detected in all infected loops at 1 h PI. The area of TUNEL-labeled cells in the wild-type infected loops was significantly higher than that of the controls at 12 h PI, when a severe inflammatory response and tissue damage had already developed. The sopB mutant induced the same amount of TUNEL-positive cells as the wild type, but it was attenuated for induction of fluid secretion and inflammation. Our results indicate that serovar Typhimurium induced cell death is not required to trigger an early inflammatory response and fluid accumulation in the ileum. PMID- 11402006 TI - In vivo visualization of bacterial colonization, antigen expression, and specific T-cell induction following oral administration of live recombinant Salmonella enterica serovar Typhimurium. AB - Live attenuated Salmonella strains that express a foreign antigen are promising oral vaccine candidates. Numerous genetic modifications have been empirically tested, but their effects on immunogenicity are difficult to interpret since important in vivo properties of recombinant Salmonella strains such as antigen expression and localization are incompletely characterized and the crucial early inductive events of an immune response to the foreign antigen are not fully understood. Here, methods were developed to directly localize and quantitate the in situ expression of an ovalbumin model antigen in recombinant Salmonella enterica serovar Typhimurium using two-color flow cytometry and confocal microscopy. In parallel, the in vivo activation, blast formation, and division of ovalbumin-specific CD4(+) T cells were followed using a well-characterized transgenic T-cell receptor mouse model. This combined approach revealed a biphasic induction of ovalbumin-specific T cells in the Peyer's patches that followed the local ovalbumin expression of orally administered recombinant Salmonella cells in a dose- and time-dependent manner. Interestingly, intact Salmonella cells and cognate T cells seemed to remain in separate tissue compartments throughout induction, suggesting a transport of killed Salmonella cells from the colonized subepithelial dome area to the interfollicular inductive sites. The findings of this study will help to rationally optimize recombinant Salmonella strains as efficacious live antigen carriers for oral vaccination. PMID- 11402007 TI - Complete DNA sequence of Yersinia enterocolitica serotype 0:8 low-calcium response plasmid reveals a new virulence plasmid-associated replicon. AB - The complete nucleotide sequence and organization of the Yersinia enterocolitica serotype 0:8 low-calcium-response (LCR) plasmid, pYVe8081, were determined. The 67,720-bp plasmid encoded all the genes known to be part of the LCR stimulon except for ylpA. Eight of 13 intact open reading frames of unknown function identified in pYVe8081 had homologues in Yersinia pestis plasmid pCD1 or in Y. enterocolitica serotype 0:9 plasmid pYVe227. A region of approximately 17 kbp showed no DNA identity to pCD1 or pYVe227 and contained six potential new genes, a possible new replicon, and two intact insertion sequence (IS) elements. One intact IS element, ISYen1, was a new IS belonging to the IS256 family. Several vestigial IS elements appeared different from the IS distribution seen in the other LCR plasmids. The RepA proteins encoded by Y. enterocolitica serotype 0:8 pYVeWA and pYVe8081 were identical. The putative pYVe8081 replicon showed significant homology to the IncL/M replicon of pMU407.1 but was only distantly related to the replicons of pCD1 and pYVe227. In contrast, the putative partitioning genes of pYVe8081 showed 97% DNA identity to the spy/sopABC loci of pCD1 and pYVe227. Sequence analysis suggests that Yersinia LCR plasmids are from a common ancestor but that Y. enterocolitica serotype 0:8 plasmid replicons may have evolved independently via cointegrate formation following a transposition event. The change in replicon structure is predicted to change the incompatibility properties of Y. enterocolitica serotype 0:8 plasmids from those of Y. enterocolitica serotype 0:9 and Y. pestis LCR plasmids. PMID- 11402008 TI - Granulomatous skin lesions in moray eels caused by a novel Mycobacterium species related to Mycobacterium triplex. AB - An outbreak of granulomatous dermatitis was investigated in a captive population of moray eels. The affected eels had florid skin nodules concentrated around the head and trunk. Histopathological examination revealed extensive granulomatous inflammation within the dermis and subcutaneous fascial plane between the fat and axial musculature. Acid-fast rods were detected within the smallest lesions, which were presumably the ones that had developed earliest. Eventually, after several months of incubation at room temperature, a very slowly growing acid-fast organism was isolated. Sequencing of the 16S rRNA gene identified it as a Mycobacterium species closely related (0.59% divergence) to M. triplex, an SAV mycobacterium. Intradermal inoculation of healthy green moray eels with this organism reliably reproduced the lesion. Experimentally induced granulomatous dermatitis appeared within 2 weeks of inoculation and slowly but progressively expanded during the 2 months of the experiment. Live organisms were recovered from these lesions at all time points, fulfilling Koch's postulates for this bacterium. In a retrospective study of tissues collected between 1993 and 1999 from five spontaneous disease cases, acid-fast rods were consistently found within lesions, and a nested PCR for the rRNA gene also demonstrated the presence of mycobacteria within affected tissues. PMID- 11402009 TI - Expression and functional properties of the Streptococcus intermedius surface protein antigen I/II. AB - Streptococcus intermedius is associated with deep-seated purulent infections. In this study, we investigated expression and functional activities of antigen I/II in S. intermedius. The S. intermedius antigen I/II appeared to be cell surface associated, with a molecular mass of approximately 160 kDa. Northern blotting indicated that the S. intermedius NCTC 11324 antigen I/II gene was transcribed as a monocistronic message. Maximum expression was seen during the early exponential phase. Insertional inactivation of the antigen I/II gene resulted in reduced hydrophobicity during early exponential phase, whereas no effect was detected during mid- and late exponential phases. Binding to human fibronectin and laminin was reduced in the isogenic mutant, whereas binding to human collagen types I and IV and to rat collagen type I was not significant for either the wild type or the mutant. Compared to the wild type, the capacity of the isogenic mutant to induce interleukin 8 (IL-8) release by THP-1 monocytic cells was significantly reduced. The results indicate that the S. intermedius antigen I/II is involved in adhesion to human receptors and in IL-8 induction. PMID- 11402010 TI - Normal IncA expression and fusogenicity of inclusions in Chlamydia trachomatis isolates with the incA I47T mutation. AB - To investigate the correlation between the incA I47T mutation in Chlamydia trachomatis and the nonfusogenic phenotype, the incA genes of 25 isolates were sequenced. Four major sequence types were identified. Seven isolates (28%) had the I47T mutation. Isolates representing the four sequence types expressed IncA in the membrane of one large single inclusion. In conclusion, the incA I47T mutation is not associated with the nonfusogenic phenotype. PMID- 11402011 TI - Variation in the infectivity of Listeria monocytogenes isolates following intragastric inoculation of mice. AB - The infectivities of 66 Listeria monocytogenes isolates were assessed by intragastric inoculation of mice. Eight were poorly infective. Serovars 4b and 1/2 were more infective than serovars 3 and 4nonb. A noninfective isolate was cleared more rapidly from the cecum than were infective isolates, suggesting that survival in the gut may relate to infectivity. PMID- 11402012 TI - Distribution of quorum-sensing genes in the Burkholderia cepacia complex. AB - The distribution of quorum-sensing genes among strains from seven genomovars of the Burkholderia cepacia complex was examined by PCR. cepR and cepI were amplified from B. cepacia genomovars I and III, B. stabilis, and B. vietnamiensis. cepR was also amplified from B. multivorans and B. cepacia genomovar VI. bviIR were amplified from B. vietnamiensis. All genomovars produced N-octanoyl-L-homoserine lactone and N-hexanoyl-L-homoserine lactone. B. vietnamiensis and B. cepacia genomovar VII produced additional N-acyl-L homoserine lactones. PMID- 11402013 TI - Expression of genes encoding Th1 cell-activating cytokines and lymphoid homing chemokines by chlamydia-pulsed dendritic cells correlates with protective immunizing efficacy. AB - We studied the expression of cytokines, chemokines, and chemokine receptors by the RNase protection assay in chlamydia-pulsed dendritic cells to better understand their potent anti-chlamydial immunizing properties. We found that chlamydia-pulsed dendritic cells express a complex profile of inflammatory and immunomodulatory molecules. These include CCR-7, interleukin-12, and interferon induced protein 10, molecules that might influence the homing of pulsed dendritic cells to the site of chlamydial infection and the induction of a local protective CD4(+) Th1 cellular immunity. PMID- 11402014 TI - In vitro and in vivo assessment of Salmonella enterica serovar Typhimurium DT104 virulence. AB - Multidrug-resistant Salmonella enterica serovar Typhimurium phage type DT104 has become a widespread cause of human and other animal infection worldwide. The severity of clinical illness in S. enterica serovar Typhimurium DT104 outbreaks has led to the suggestion that this strain possesses enhanced virulence. In the present study, in vitro and in vivo virulence-associated phenotypes of several clinical isolates of S. enterica serovar Typhimurium DT104 were examined and compared to S. enterica serovar Typhimurium ATCC 14028s. The ability of these DT104 isolates to survive within murine peritoneal macrophages, invade cultured epithelial cells, resist antimicrobial actions of reactive oxygen and nitrogen compounds, and cause lethal infection in mice were assessed. Our results failed to demonstrate that S. enterica serovar Typhimurium DT104 isolates are more virulent than S. enterica serovar Typhimurium ATCC 14028s. PMID- 11402015 TI - Dr operon-associated invasiveness of Escherichia coli from pregnant patients with pyelonephritis. AB - We used a gentamicin protection assay to assess the ability of gestational pyelonephritis isolates of Escherichia coli to invade HeLa cells. The ability to enter HeLa cells was strongly associated with the presence of Dr operons coding for Dr adhesins. In contrast, the nonivasive isolates predominantly expressed papG, coding for P fimbriae. PMID- 11402016 TI - Vibrio cholerae tolC is required for bile resistance and colonization. AB - TolC and its homologues are outer membrane proteins that are essential for the transport of many molecules across the cell envelope. In this study we characterized the gene encoding Vibrio cholerae TolC. V. cholerae tolC mutants failed to secrete the RTX cytotoxin, were hypersensitive to antimicrobial agents, and were deficient in intestinal colonization. PMID- 11402017 TI - Development of a gene inactivation system for Bacteroides forsythus: construction and characterization of a BspA mutant. AB - Bacteroides forsythus is a gram-negative anaerobic bacterium associated with periodontitis. The bspA gene encoding a cell surface associated leucine-rich repeat protein (BspA) involved in adhesion to fibronectin and fibrinogen was recently cloned from this bacterium in our laboratory. We now describe the construction of a BspA-defective mutant of B. forsythus. This is the first report describing the generation of a specific gene knockout mutant of B. forsythus, and this procedure should be useful in establishing the identity of virulence associated factors in these organisms. PMID- 11402018 TI - Francisella tularensis induces cytopathogenicity and apoptosis in murine macrophages via a mechanism that requires intracellular bacterial multiplication. AB - The murine macrophage-like cell line J774.A1 ingests and allows intracellular growth of Francisella tularensis. We demonstrate that, after 24 h of infection, a pronounced cytopathogenicity resulted and the J774 cells were undergoing apoptosis. Despite this host cell apoptosis, no decrease in bacterial numbers was observed. When internalization of bacteria was prevented or intracellularly located F. tularensis bacteria were eradicated within 12 h, the progression of host cell cytopathogenicity and apoptosis was prevented. PMID- 11402019 TI - Hyphae and yeasts of Candida albicans differentially regulate interleukin-12 production by human blood monocytes: inhibitory role of C. albicans germination. AB - The role of Candida albicans yeast-to-hyphae transition in interleukin-12 (IL-12) production by monocytes was investigated. Germinating C. albicans not only failed to induce IL-12 p70 but also suppressed IL-12 production induced by heat-killed C. albicans. Comparison of the abilities of germinating C. albicans and agerminating mutants to inhibit IL-12 production showed that germination of C. albicans plays a critical role in the inhibition of IL-12 production. PMID- 11402020 TI - Synthetic polysaccharide type 3-related di-, tri-, and tetrasaccharide-CRM(197) conjugates induce protection against Streptococcus pneumoniae type 3 in mice. AB - Di-, tri-, and tetrasaccharides, synthesized according to the chemical structure of pneumococcal polysaccharide type 3 (PS3), were coupled to the cross-reactive material (CRM(197)) of modified diphtheria toxin in different molar carbohydrate/protein ratios using the squarate coupling method. To study protective immunity, female BALB/c mice were subcutaneously immunized twice (with a 3-week interval) using the amount of conjugates corresponding to 2.5 microg of oligosaccharide per mouse. The conjugates evoked PS3 binding immunoglobulin G antibodies that lasted for at least 7 weeks after the booster. Immunogenicity was not influenced by the carbohydrate/protein ratio. All mice with PS3-specific antibodies survived the intraperitoneal challenge with Streptococcus pneumoniae type 3. Therefore, synthetic oligosaccharide-protein conjugates might have potential as vaccines. PMID- 11402021 TI - Characterization of an N-terminally truncated cyclin A isoform in mammalian cells. AB - Cyclin A is essential for regulating key transitions in the eukaryotic cell cycle including initiation of DNA replication and mitosis. This paper describes the characterization of a truncated cyclin A isoform (cyclin A(t)) in vitro in cultured mammalian cells and in mouse tissues. The presence of cyclin A(t) in specific cell types correlates with the ability of cell extracts to cleave in vitro translated cyclin A. In CHO-K1 cells, cyclin A processing to cyclin A(t) occurs at the N terminus; it does not involve the 26 S proteasome, nor could it be induced by conditional overexpression of the cyclin-dependent kinase inhibitor p27(Kip1). However, high cell densities lead to increased cyclin A(t) levels. Unlike full-length cyclin A, cyclin A(t) localizes to the cytoplasm, where it binds Cdk2. The data suggest that cyclin A processing occurs in vivo to yield an N-terminally truncated isoform by an unknown mechanism that is regulated by cell density. Differential subcellular localization may provide the first insights into the physiological role of cyclin A(t). PMID- 11402022 TI - The crystal structure of the MJ0796 ATP-binding cassette. Implications for the structural consequences of ATP hydrolysis in the active site of an ABC transporter. AB - The crystal structure of the MJ0796 ATP-binding cassette, a member of the o228/LolD transporter family, has been determined at 2.7-A resolution with MgADP bound at its active site. Comparing this structure with that of the ATP-bound form of the HisP ATP-binding cassette (Hung, L. W., Wang, I. X., Nikaido, K., Liu, P. Q., Ames, G. F., and Kim, S. H. (1998) Nature 396, 703-707) shows a 5-A withdrawal of a phylogenetically invariant glutamine residue from contact with the gamma-phosphate of ATP in the active site. This glutamine is located in a protein segment that links the rigid F(1)-type ATP-binding core of the enzyme to an ABC transporter-specific alpha-helical subdomain that moves substantially away from the active site in the MgADP-bound structure of MJ0796 compared with the ATP bound structure of HisP. A similar conformational effect is observed in the MgADP bound structure of MJ1267 (Karpowich, N., et al. (2001) Structure, in press), establishing the withdrawal of the glutamine and the coupled outward rotation of the alpha-helical subdomain as consistent consequences of gamma-phosphate release from the active site of the transporter. Considering this subdomain movement in the context of a leading model for the physiological dimer of cassettes present in ABC transporters indicates that it produces a modest mechanical change that is likely to play a role in facilitating nucleotide exchange out of the ATPase active site. Finally, it is noteworthy that one of the intersubunit packing interactions in the MJ0796 crystal involves antiparallel beta-type hydrogen bonding interactions between the outermost beta-strands in the two core beta sheets, leading to their fusion into a single extended beta-sheet, a type of structural interaction that has been proposed to play a role in mediating the aggregation of beta-sheet-containing proteins. PMID- 11402023 TI - Parathyroid hormone-related peptide stimulates osteogenic cell proliferation through protein kinase C activation of the Ras/mitogen-activated protein kinase signaling pathway. AB - We investigated the mechanisms of parathyroid hormone-related peptide (PTHrP) mediated effects on osteogenic cells in primary rat bone marrow cell (BMC) cultures. We first demonstrated by reverse transcriptase-polymerase chain reaction and immunocytochemistry that BMCs express the type I parathyroid hormone/PTHrP receptor. Treatment with PTHrP increased osteogenic cell proliferation as determined by [(3)H]thymidine and bromodeoxyuridine incorporation and augmented osteogenic colonies. Immunocytochemistry and Western blotting revealed no direct effect on expression of the osteoblast markers, type I collagen, bone sialoprotein, and osteocalcin, indicating that PTHrP did not directly stimulate differentiation in this system. PTHrP increased mitogen activated protein kinase (MAPK) activity in BMC and MAPK activity, and PTHrP induced osteogenic cell proliferation could be blocked by the MEK inhibitor PD 098059. PTHrP also increased Ras activity in BMC. Although wortmannin and H8, inhibitors of phosphoinositol 3-kinase and protein kinase A, respectively, did not block PTHrP-stimulated Ras or MAPK activity, chelerythrin chloride, a known protein kinase C inhibitor, did block these PTHrP actions as well as PTHrP induced osteogenic cell proliferation. These results demonstrate that PTHrP stimulates osteogenic cell proliferation in rat marrow mesenchymal progenitor cells through protein kinase C-dependent activation of the Ras and MAPK signaling pathway. PMID- 11402024 TI - Caspase-dependent cleavage of ErbB-2 by geldanamycin and staurosporin. AB - The geldanamycin-induced degradation of ErbB-2 produces a 23-kDa carboxyl terminal fragment, which has been isolated and subjected to amino-terminal microsequencing. The obtained sequence indicates that the amino terminus of this fragment corresponds to Gly-1126 of ErbB-2. Analysis of the residues immediately before Gly-1126 suggests that cleavage may involve caspase activity. Site directed mutagenesis of Asp-1125 in ErbB-2 prevents geldanamycin-provoked formation of the 23-kDa fragment, consistent with the requirement of this residue for caspase-dependent cleavage in known substrates. Also, the addition of the pan caspase inhibitor Z-VAD-FMK blocks formation of the 23-kDa ErbB-2 fragment in cells exposed to geldanamycin. Interestingly, staurosporin and curcumin are also shown to provoke the degradation of ErbB-2 with formation of the 23-kDa carboxyl terminal fragment. The generation of this fragment by staurosporin or curcumin is likewise blocked by caspase inhibition. Caspase inhibition does not prevent accelerated degradation of the 185-kDa native ErbB-2 in geldanamycin-treated cells but does significantly prevent staurosporin-stimulated metabolic loss of ErbB-2. PMID- 11402025 TI - Inhibition of proprotein convertases is associated with loss of growth and tumorigenicity of HT-29 human colon carcinoma cells: importance of insulin-like growth factor-1 (IGF-1) receptor processing in IGF-1-mediated functions. AB - Proprotein convertases (PCs) of the subtilisin/kexin family are responsible for the activation of prohormones, protrophic factors, and their receptors. We sought to determine whether loss of PC-mediated activities might affect the malignant phenotypes of cancer cells. Stable transfectants of alpha(1)-antitrypsin Portland (alpha(1)-PDX) cDNA, coding for a potent PC inhibitor, were analyzed in model HT 29 cells (HT-29/PDX) and in other cell lines. Expression of alpha(1)-PDX resulted in a proinsulin-like growth factor-1 receptor (pro-IGF-1R) processing blockade, hence inhibiting the ability of exogenous IGF-1 to induce tyrosine phosphorylation of its beta-subunit and insulin-related substrate-1. Coexpression of IGF-1R with four different PCs or the novel convertase SKI-1 in the furin defective LoVo-C5 cells demonstrated that pro-IGF-1R ( approximately 200 kDa) cleavage into IGF-1R (beta-subunit, approximately 105 kDa) can be achieved by furin and PC5A, but not by PACE4, PC7, or SKI-1. Expression of alpha(1)-PDX resulted in reduction of DNA synthesis and in anchorage-independent growth. Following serum deprivation, the alpha(1)-PDX transfectants exhibited an enhanced apoptotic phenotype and were insensitive to IGF-1-mediated [(3)H]thymidine incorporation and protection against apoptosis. These cells showed reduced invasiveness that paralleled decreased mRNA levels of urokinase-type plasminogen activator and its receptor, tissue-type plasminogen activator, and plasminogen activator inhibitor-1. Comparative subcutaneous inoculation of cells in nude mice revealed that animals injected with HT-29/PDX cells exhibited delayed and lower incidence of tumor development as well as reduced tumor size. Immunohistochemical analysis of CD31 antigen expression, a marker of endothelial cells, revealed reduced HT-29/PDX tumor vascularization. These findings indicate that PCs actively contribute to the growth and malignant phenotypes of HT-29 tumors, suggesting that PC inhibition strategies may be a useful adduct to the arsenal of colorectal anticancer gene therapies. PMID- 11402026 TI - In vivo detection of hetero-association of glycophorin-A and its mutants within the membrane. AB - Protein recognition within the membrane is a crucial process for numerous biological activities. Detection of such interaction is limited because of difficulties that arise from the hydrophobic environment of the membrane. We detected direct hetero-oligomerization of the glycophorin-A (GPA) transmembrane segments in vivo through inhibition of ToxR transcription activator dimer formation. We investigated the amino acids important for hetero-oligomerization within the membrane, using peptide analog segments of the transmembrane domain of glycophorin A. The wild type ([WT]GPA) and alanine mutant ([A]GPA) were able to interfere with and inhibit the proper dimerization of the ToxR-GPA transcription factor. Conversely, a second alanine mutant ([A(2)]GPA), a glycine mutant ([G]GPA), and a scrambled analog ([SC]GPA) were virtually inactive. Binding studies reveal similar membrane partitions for [WT]GPA, [G]GPA, and [SC]GPA, whereas membrane partition of [A]GPA and [A(2)]GPA are lower. Spectral analysis of fluorescent-labeled analogs revealed a significant blue shift, indicating membrane insertion. Our results suggest that the GXXXG motif, found in homo oligomerization, is not sufficient for hetero-oligomerization in a biological membrane, whereas an extended motif, LIXXGXXXGXXXT, is sufficient. Interfering with hetero-oligomerization within the membrane can be a useful strategy for characterizing such interactions and possibly modulating membrane protein activity. PMID- 11402027 TI - c-Myc-mediated regulation of telomerase activity is disabled in immortalized cells. AB - Myc overexpression is a hallmark of human cancer and promotes transformation by facilitating immortalization. This function has been linked to the ability of c Myc to induce the expression of the catalytic subunit of telomerase, telomerase reverse transcriptase (TERT), as ectopic expression of TERT immortalizes some primary human cell types. c-Myc up-regulates telomerase activity in primary mouse embryonic fibroblasts (MEFs) and myeloid cells. Paradoxically, Myc overexpression also triggers the ARF-p53 apoptotic program, which is activated when MEFs undergo replicative crises following culture ex vivo. The rare immortal variants that arise from these cultures generally suffer mutations in p53 or delete Ink4a/ARF, and Myc greatly increases the frequency of these events. Alternative reading frame (ARF)- and p53-null MEFs have increased telomerase activity, as do variant immortal clones that bypass replicative crisis. Similarly, immortal murine NIH 3T3 fibroblasts and myeloid 32D.3 and FDC-P1.2 cells do not express ARF and have robust telomerase activity. However, Myc overexpression in these immortal cells results in remarkably discordant regulation of TERT and telomerase activity. Furthermore, in MEFs and 32D.3 cells TERT expression and telomerase activity are regulated independently of endogenous c-Myc. Thus, the regulation of TERT and telomerase activity is complex and is also regulated by factors other than Myc, ARF, or p53. PMID- 11402028 TI - Lipopolysaccharide induces Rac1-dependent reactive oxygen species formation and coordinates tumor necrosis factor-alpha secretion through IKK regulation of NF kappa B. AB - Reactive oxygen species (ROS) are important second messengers generated in response to many types of environmental stress. In this setting, changes in intracellular ROS can activate signal transduction pathways that influence how cells react to their environment. In sepsis, a dynamic proinflammatory cellular response to bacterial toxins (e.g. lipopolysaccharide or LPS) leads to widespread organ damage and death. The present study demonstrates for the first time that the activation of Rac1 (a GTP-binding protein), and the subsequent production of ROS, constitutes a major pathway involved in NFkappaB-mediated tumor necrosis factor-alpha (TNFalpha) secretion following LPS challenge in macrophages. Expression of a dominant negative mutant of Rac1 (N17Rac1) reduced Rac1 activation, ROS formation, NFkappaB activation, and TNFalpha secretion following LPS stimulation. In contrast, expression of a dominant active form of Rac1 (V12Rac1) mimicked these effects in the absence of LPS stimulation. IKKalpha and IKKbeta were both required downstream modulators of LPS-activated Rac1, since the expression of either of the IKK dominant mutants (IKKalphaKM or IKKbetaKA) drastically reduced NFkappaB-dependent TNFalpha secretion. Moreover, studies using CD14 blocking antibodies suggest that Rac1 induces TNFalpha secretion through a pathway independent of CD14. However, a maximum therapeutic inhibition of LPS-induced TNFalpha secretion occurred when both CD14 and Rac1 pathways were inhibited. Our results suggest that targeting both Rac1- and CD14-dependent pathways could be a useful therapeutic strategy for attenuating the proinflammatory cytokine response during the course of sepsis. PMID- 11402029 TI - Bipartite binding of a kinase activator activates Cdc7-related kinase essential for S phase. AB - Dfp1/Him1 protein of fission yeast, Schizosaccharomyces pombe, encodes the regulatory subunit for Hsk1 kinase, a homologue of budding yeast Cdc7 kinase essential for initiation and progression of the S phase of the cell cycle. This protein binds and activates Hsk1 kinase, which phosphorylates the MCM2 protein. Comparison of the amino acid sequences of the Cdc7 regulatory subunits from various eukaryotes revealed the presence of three small stretches of conserved amino acid sequences, namely Dbf4 motifs N, M, and C. We report here that the Dbf4 motif M, a unique proline-rich motif, and the Dbf4 motif C, a C(2)H(2)-type zinc finger motif, are essential for mitotic functions of Dfp1/Him1 protein as well as for full-level activation of Hsk1 kinase. In vitro, a small segment containing the Dbf4 motif M or C alone binds to and partially activates Hsk1. Co expression of these two segments augments the extent of activation. Furthermore, a fused polypeptide containing only Dbf4 motifs M and C without any spacer can activate Hsk1 and is capable of rescuing the growth defect of him1 null cells. Insertion of a long stretch of amino acids between the motif M and motif C can be tolerated for mitotic functions. On the other hand, internal deletion of Dbf4 motif N, which has some similarity with the BRCA C-terminal domain motif, results in a defect in hydroxyurea-induced checkpoint responses and sensitivity to methyl methane sulfonate, yet mitotic functions and kinase activation are intact. In one hybrid assays with budding yeast Dbf4, motif N mutants exhibit reduced interaction with a replication origin. Our observations suggest the molecular architecture of Cdc7.Dbf4-related kinase complexes at the origins, in which they are tethered to replication machinery through Dbf4 motif N and the catalytic subunits are activated through bipartite binding of Dbf4 motifs M and C of the regulatory subunits. PMID- 11402030 TI - Chloroplasts have a novel Cpn10 in addition to Cpn20 as co-chaperonins in Arabidopsis thaliana. AB - Previously, we characterized a mitochondrial co-chaperonin (Cpn10) and a chloroplast co-chaperonin (Cpn20) from Arabidopsis thaliana (Koumoto, Y., Tsugeki, R., Shimada, T., Mori, H., Kondo, M., Hara-Nishimura, I., and Nishimura, M. (1996) Plant J. 10, 1119-1125; Koumoto, Y., Shimada, T., Kondo, M., Takao, T., Shimonishi, Y., Hara-Nishimura, I., and Nishimura, M. (1999) Plant J. 17, 467 477). Here, we report a third co-chaperonin. The cDNA was 603 base pairs long, encoding a protein of 139 amino acids. From a sequence analysis, the protein was predicted to have one Cpn10 domain with an amino-terminal extension that might work as a chloroplast transit peptide. This novel Cpn10 was confirmed to be localized in chloroplasts, and we refer to it as chloroplast Cpn10 (chl-Cpn10). The phylogenic tree that was generated with amino acid sequences of other co chaperonins indicates that chl-Cpn10 is highly divergent from the others. In the GroEL-assisted protein folding assay, about 30% of the substrates were refolded with chl-Cpn10, indicating that chl-Cpn10 works as a co-chaperonin. A Northern blot analysis revealed that mRNA for chl-Cpn10 is accumulated in the leaves and stems, but not in the roots. In germinating cotyledons, the accumulation of chl Cpn10 was similar to that of chloroplastic proteins and accelerated by light. It was proposed that two kinds of co-chaperonins, Cpn20 and chl-Cpn10, work independently in the chloroplast. PMID- 11402031 TI - Human DNA polymerase iota promiscuous mismatch extension. AB - Human DNA polymerase iota is a low-fidelity template copier that preferentially catalyzes the incorporation of the wobble base G, rather than the Watson-Crick base A, opposite template T (Tissier, A., McDonald, J. P., Frank, E. G., and Woodgate, R. (2000) Genes Dev. 14, 1642-1650; Johnson, R. E., Washington, M. T., Haracska, L., Prakash, S., and Prakash, L. (2000) Nature 406, 1015-1019; Zhang, Y., Yuan, F., Wu, X., and Wang, Z. (2000) Mol. Cell. Biol. 20, 7099-7108). Here, we report on its ability to extend all 12 possible mispairs and 4 correct pairs in different sequence contexts. Extension from both matched and mismatched primer termini is generally most efficient and accurate when A is the next template base. In contrast, extension occurs less efficiently and accurately when T is the target template base. A striking exception occurs during extension of a G:T mispair, where the enzyme switches specificity, "preferring" to make a correct A:T base pair immediately downstream from an originally favored G:T mispair. Polymerase iota generates a variety of single and tandem mispairs with high frequency, implying that it may act as a strong mutator when copying undamaged DNA templates in vivo. Even so, its limited ability to catalyze extension from a relatively stable primer/template containing a "buried" mismatch suggests that polymerase iota-catalyzed errors are confined to short template regions. PMID- 11402032 TI - pEg2 aurora-A kinase, histone H3 phosphorylation, and chromosome assembly in Xenopus egg extract. AB - In eukaryotes cell division is accompanied by phosphorylation of histone H3 at serine 10. In this work we have studied the kinase activity responsible for this histone H3 modification by using cell-free extracts prepared from Xenopus eggs. We have found that the Xenopus aurora-A kinase pEg2, immunoprecipitated from the extract, is able to phosphorylate specifically histone H3 at serine 10. The enzyme is incorporated into chromatin during in vitro chromosome assembly, and the kinetics of this incorporation parallels that of histone H3 phosphorylation. Recombinant pEg2 phosphorylates efficiently histone H3 at serine 10 in reconstituted nucleosomes and in sperm nuclei decondensed in heated extracts. These data identify pEg2 as a good candidate for mitotic histone H3 kinase. However, immunodepletion of pEg2 does not interfere with the chromosome assembly properties of the extract nor with the pattern of H3 phosphorylation, suggesting the existence of multiple kinases involved in this H3 modification in Xenopus eggs. This hypothesis is supported by in gel activity assay experiments using extracts from Saccharomyces cerevisiae. PMID- 11402033 TI - Modification of the beta 2-adrenergic receptor to engineer a receptor-effector complex for gene therapy. AB - Depressed G-protein-coupled receptor (GPCR) signaling has been implicated as a component of the pathophysiology of a number of complex diseases including heart failure and asthma, and augmentation or restoration of signaling by various means has been shown to improve organ function. Because some properties of native GPCRs are disadvantageous for ectopic therapeutic expression, we utilized the beta(2) adrenergic receptor (beta(2)AR) as a scaffold to construct a highly modified therapeutic receptor-effector complex (TREC) suitable for gene therapy. Altogether, 19 modifications were made to the receptor. The ligand-binding site was re-engineered in TM-3 so that a beta-hydroxylmethyl side chain acts as a proton donor for the binding of a novel ligand. In addition, sites critical for agonist-promoted down-regulation in the amino terminus and for phosphorylation by GPCR kinases, and protein kinases A and C, in the third intracellular loop and the carboxyl terminus of the receptor were altered. These modifications of the receptor resulted in depressed agonist-stimulated adenylyl cyclase activity (26.8 +/- 2.1 versus 41.4 +/- 8 pmol/min/mg for wild-type beta(2)AR). This was fully restored by fusing the carboxyl terminus of the modified receptor to G alpha(s) (43.3 +/- 2.7 pmol/min/mg). The fully modified fused receptor was not activated by beta-agonists but rather by a nonbiogenic amine agonist that itself failed to activate the wild-type beta(2)AR. This two-way selectivity thus provides targeted activation based on physiologic status. Furthermore, the TREC did not display tachyphylaxis to prolonged agonist exposure (desensitization was 1 +/- 5% versus 55 +/- 4% for wild-type beta(2)AR). Thus, despite extensive alterations in regions of conformational lability, the beta(2)AR can be tailored to have optimal signaling characteristics for gene therapy. As a general paradigm, TRECs for enhancement of other G-protein signaling appear to be feasible for modification of other pathologic states. PMID- 11402034 TI - Mapping the functional domains of HAP95, a protein that binds RNA helicase A and activates the constitutive transport element of type D retroviruses. AB - The complex retroviruses such as human immunodeficiency virus, type 1, employ a virally encoded protein, Rev, to mediate the nuclear export of unspliced and partially spliced mRNA. In contrast, the simian type D retroviruses act through a cis-acting constitutive transport element (CTE) that presumably interacts directly with cellular export proteins. We first reported that RNA helicase A (RHA) is a shuttle protein that binds to functional CTE in vitro and in vivo. Recently, we isolated a novel protein, HAP95, that specifically binds to the nuclear transport domain of RHA and up-regulates CTE-mediated gene expression. Here, using truncation and deletion mutations, we mapped the domains of HAP95 that are important for RHA binding, transactivation of CTE, and nuclear cytoplasmic shuttling. We report evidence for a novel nuclear export signal in HAP95 and showed that the domains involved in RHA binding and nuclear localization are required for CTE activation. Finally, we showed that HAP95 synergizes significantly with RHA on CTE-mediated reporter gene expression and promotes nuclear export of unspliced mRNA in transfected cells. Taken together, these data support the proposal that HAP95 specifically facilitates CTE-mediated gene expression by directly binding to RHA. PMID- 11402035 TI - Glycogen synthase kinase-3beta regulates presenilin 1 C-terminal fragment levels. AB - The majority of familial Alzheimer's disease cases have been attributed to mutations in the presenilin 1 (PS1) gene. PS1 is synthesized as an inactive holoprotein that undergoes endoproteolytic processing to generate a functional N- and C-terminal heterodimer (NTF and CTF, respectively). We identified a single residue in PS1, Ser(397), which regulates the CTF levels in a population of dimer that has a rapid turnover. This residue is part of a highly conserved glycogen synthase kinase-3beta (GSK-3beta) consensus phosphorylation site within the loop domain of PS1. Site-directed mutagenesis at the Ser(397) position increased levels of PS1 CTF but not NTF or holoprotein. Similar increases in only CTF levels were seen when cells expressing wild type PS1 were treated with lithium chloride, an inhibitor of GSK-3beta. Both wild type and PS1 S397A CTF displayed a biphasic turnover, reflecting rapidly degraded and stable populations. Rapid turnover was delayed for mutant PS1 S397A, causing increased CTF. These data demonstrate that PS1 NTF.CTF endoproteolytic fragments are generated in excess, that phosphorylation at Ser(397) by GSK-3beta regulates the discard of excess CTF, and that the disposal of surplus NTF is mediated by an independent mechanism. Overall, the results indicate that production of active NTF.CTF dimer is more complex than limited endoproteolysis of PS1 holoprotein and instead involves additional regulatory events. PMID- 11402036 TI - DNA synthesis by HIV-1 reverse transcriptase at the central termination site: a kinetic study. AB - Human immunodeficiency virus, type 1 (HIV-1) reverse transcriptase (RT) terminates plus-strand DNA synthesis at the center of the HIV-1 genome, a process important for HIV-1 infectivity. The central termination sequence contains two termination sites (Ter1 and Ter2) located at the 3'-end of A(n)T(m) motifs, and the narrowing of the DNA minor groove generated by these motifs is responsible for termination. Kinetic data associated with the binding of RT and its ability to elongate in vitro various DNA duplexes and triplexes surrounding the Ter2 terminator were analyzed using a simple kinetic scheme. At Ter2, RT still displays a reasonable affinity for the corresponding DNA, but the binding of the next nucleotide and above all its incorporation rate are markedly hampered. Features affecting the width of the minor groove act directly at this last step. The constraint exerted against elongation by the A(n)T(m) tract persists at two positions downstream of the terminator. PMID- 11402037 TI - Structures of complexes formed by HIV-1 reverse transcriptase at a termination site of DNA synthesis. AB - This study presents structural parameters associated with termination of human immunodeficiency virus, type 1 (HIV-1) reverse transcriptase (RT) at Ter2, the major termination site located in the center of the HIV-1 genome. DNA footprinting studies of various elongation complexes formed by RT around wild type and mutant Ter2 sites have revealed two major structural transformations of these complexes when the enzyme gets closer to Ter2. First, the interactions between RT and the DNA duplex are less extended, although the global affinity of the enzyme for this duplex is only decreased by 2-fold. Second, there is an atypical positioning of the RT RNase H domain on the DNA duplex. We interpret our data as indicating that the A(n)T(m) motif located upstream of Ter2 prevents a classical positioning of the enzyme on the double-stranded part of the DNA duplex at some precise positions of elongation downstream of this motif. Instead, novel species of binary and/or ternary complexes, characterized by atypical footprints, are formed. The new rate-limiting step of the reaction, characterized in the preceding paper (Lavigne, M., Polomack, L., and Buc, H. (2001) J. Biol. Chem. 276, 31429-31438), would be a transition leading from these new species to a catalytically competent ternary complex. PMID- 11402038 TI - Recruitment of the male-specific lethal (MSL) dosage compensation complex to an autosomally integrated roX chromatin entry site correlates with an increased expression of an adjacent reporter gene in male Drosophila. AB - Drosophila dosage compensate (equalize X-linked gene products) by doubling the transcription of most X-linked genes in males. The MSL (male-specific lethal) ribonucleoprotein complex consisting of at least five proteins and two non-coding RNAs (roX1 and roX2) is essential for this transcription response. Recently it has been shown that the X-linked roX1 and roX2 genes each contain at least one chromatin entry site for the MSL complex. In this study we show that insertion of either roX1 or roX2 DNA sequences, upstream of an insulated lacZ reporter gene controlled with the constitutive armadillo promoter (arm-lacZ), results in a significant elevation of expression of lacZ in males. However, full compensation, that is a precise doubling of lacZ expression in males relative to females, was only observed in some lines carrying autosomal insertions of either roX1-arm-lacZ or roX2-arm-lacZ transgenes. Furthermore, we found that a 419-base pair fragment of roX1 that contains an MSL binding site was sufficient to cause a modest elevation of expression of lacZ in males, but this response was significantly less than obtained with a full-length roX1 cDNA. This is the first direct demonstration that insertion of an MSL chromatin entry site on an autosome results in elevated expression in males of genes near the entry site. PMID- 11402039 TI - Cloning and characterization of three Xenopus slug promoters reveal direct regulation by Lef/beta-catenin signaling. AB - In amphibians and birds, one of the first steps of neural crest cell (NCC) determination is expression of the transcription factor Slug. This marker has been used to demonstrate that BMP and Wnt molecules play a major role in NCC induction. However, it is unknown whether Slug expression is directly or indirectly regulated by these signals. We report here the cloning and characterization of three Xenopus Slug promoters: that of the Xenopus tropicalis slug gene and those of two Xenopus laevis Slug pseudoalleles. Although the three genes encode proteins with almost identical amino acid sequences and are expressed with similar spatiotemporal patterns, their 5'-flanking regions are quite different. A striking difference is a deletion in the X. tropicalis gene located precisely at the transcription initiation site that results in the X. tropicalis promoter being inefficient in X. laevis. Additionally, we identified two regions common to the three promoters that are necessary and sufficient to drive specific expression in NCCs. Interestingly, one of the common regulatory regions presents a functional Lef/beta-catenin-binding site necessary for specific expression. As the Lef.beta-catenin complex is a downstream effector of Wnt signaling, these results suggest that Xenopus Slug is a direct target of NCC determination signals. PMID- 11402040 TI - Endocytosed HSP60s use toll-like receptor 2 (TLR2) and TLR4 to activate the toll/interleukin-1 receptor signaling pathway in innate immune cells. AB - Heat shock proteins (HSPs) require no adjuvant to confer immunogenicity to bound peptides, as if they possessed an intrinsic "danger" signature. To understand the proinflammatory nature of HSP, we analyzed signaling induced by human and chlamydial HSP60. We show that both HSP60s activate the stress-activated protein kinases p38 and JNK1/2, the mitogen-activated protein kinases ERK1/2, and the I kappaB kinase (IKK). Activation of JNK and IKK proceeds via the Toll/IL-1 receptor signaling pathway involving MyD88 and TRAF6. Human fibroblasts transfected with TLR2 or TLR4 plus MD-2 gain responsiveness to HSP60, while TLR2- or TLR4-defective cells display impaired responses. Initiation of signaling requires endocytosis of HSP60 that is effectively inhibited by serum component(s). The results revealed that adjuvanticity of HSP60 operates similar to that of classical pathogen-derived ligands. PMID- 11402041 TI - Proteomic analysis of the mammalian mitochondrial ribosome. Identification of protein components in the 28 S small subunit. AB - The mammalian mitochondrial ribosome (mitoribosome) has a highly protein-rich composition with a small sedimentation coefficient of 55 S, consisting of 39 S large and 28 S small subunits. In the previous study, we analyzed 39 S large subunit proteins from bovine mitoribosome (Suzuki, T., Terasaki, M., Takemoto Hori, C., Hanada, T., Ueda, T., Wada, A., and Watanabe, K. (2001) J. Biol. Chem. 276, 21724-21736). The results suggested structural compensation for the rRNA deficit through proteins of increased molecular mass in the mitoribosome. We report here the identification of 28 S small subunit proteins. Each protein was separated by radical-free high-reducing two-dimensional polyacrylamide gel electrophoresis and analyzed by liquid chromatography/mass spectrometry/mass spectrometry using electrospray ionization/ion trap mass spectrometer to identify cDNA sequence by expressed sequence tag data base searches in silico. Twenty one proteins from the small subunit were identified, including 11 new proteins along with their complete cDNA sequences from human and mouse. In addition to these proteins, three new proteins were also identified in the 55 S mitoribosome. We have clearly identified a mitochondrial homologue of S12, which is a key regulatory protein of translation fidelity and a candidate for the autosomal dominant deafness gene, DFNA4. The apoptosis-related protein DAP3 was found to be a component of the small subunit, indicating a new function for the mitoribosome in programmed cell death. In summary, we have mapped a total of 55 proteins from the 55 S mitoribosome on the two-dimensional polyacrylamide gels. PMID- 11402042 TI - The negative effects of bile acids and tumor necrosis factor-alpha on the transcription of cholesterol 7alpha-hydroxylase gene (CYP7A1) converge to hepatic nuclear factor-4: a novel mechanism of feedback regulation of bile acid synthesis mediated by nuclear receptors. AB - Bile acids regulate the cholesterol 7alpha-hydroxylase gene (CYP7A1), which encodes the rate-limiting enzyme in the classical pathway of bile acid synthesis. Here we report a novel mechanism whereby bile acid feedback regulates CYP7A1 transcription through the nuclear receptor hepatocyte nuclear factor-4 (HNF-4), which binds to the bile acid response element (BARE) at nt -149/-118 relative to the transcription start site. Using transient transfection assays of HepG2 cells with Gal4-HNF-4 fusion proteins, we show that chenodeoxycholic acid (CDCA) dampened the transactivation potential of HNF-4. Overexpression of a constitutive active form of MEKK1, an upstream mitogen-activated protein kinase (MAPK) module triggered by stress signals, strongly repressed the promoter activity of CYP7A1 via the consensus sequence for HNF-4 embedded in the BARE. Similarly, MEKK1 inhibited the activity of HNF-4 in the Gal4-based assay. The involvement of the MEKK1-dependent pathway in the bile acid-mediated repression of CYP7A1 was confirmed by co-transfecting a dominant negative form of the stress-activated protein kinase kinase, SEK, which abolished the effect of CDCA upon CYP7A1 transcription. Treatment of transfected HepG2 cells with tumor necrosis factor alpha (TNF-alpha), an activator of the MEKK1 pathway, led to the repression of CYP7A1 via the HNF-4 site in the BARE. TNF-alpha also inhibited the transactivation potential of HNF-4. Collectively, our results demonstrate for the first time that HNF-4, in combination with a MAPK signaling pathway, acts as a bile acid sensor in the liver. Furthermore, the effects of CDCA and TNF-alpha converge to HNF-4, which binds to the BARE of CYP7A1, suggesting a link between the cascades elicited by bile acids and pro-inflammatory stimuli in the liver. PMID- 11402043 TI - Inhibition of p70(S6) kinase during transforming growth factor-beta 1/vitamin D(3)-induced monocyte differentiation of HL-60 cells allows tumor necrosis factor alpha to stimulate plasminogen activator inhibitor-1 synthesis. AB - We investigated intracellular mechanisms involved in the up-regulation of plasminogen activator inhibitor I (PAI-1) synthesis by human recombinant tumor necrosis factor-alpha (TNF) during monocyte differentiation of HL-60 cells triggered by the transforming growth factor-beta1/vitamin D(3) (TGF/D3) mixture. TGF/D3-treated cells expressed surface monocytic markers and produced noticeable amounts of PAI-1 but stopped to proliferate. A reduced p70 S6 kinase (p70(S6K)) phosphorylation was also observed and, in this situation, TNF dramatically enhanced PAI-1 synthesis. Similarly, TNF significantly up-regulated PAI-1 synthesis when p70(S6K) phosphorylation was inhibited by rapamycin. This phenomenon was not due to a general decrease in protein synthesis but involved the activation of gene transcription rather than PAI-1 mRNA stabilization. The level of the transcriptional regulator factor E2F1, a repressor of PAI-1 gene expression, was shown to be down-modulated in TGF/D3- as well as in rapamycin treated cells. Furthermore, the apoptotic effect of TNF in HL-60 cells appeared to be prevented by the addition of either TGF/D3 or rapamycin. In conclusion, these results indicate that inhibition of p70(S6K) phosphorylation during TGF/D3 induced monocyte differentiation of HL-60 cells is a determinant factor that allows TNF to exert its up-regulating effect on PAI-1 synthesis while protecting cells from apoptosis. PMID- 11402044 TI - Polar residues of the second transmembrane domain influence cation permeability of the ATP-gated P2X(2) receptor. AB - P2X receptors are simple polypeptide channels that mediate fast purinergic depolarizations in both nerve and muscle. Although the depolarization results mainly from the influx of Na(+), these channels also conduct a significant Ca(2+) current that is large enough to evoke transmitter release from presynaptic neurons. We sought to determine the molecular basis of this Ca(2+) conductance by a mutational analysis of recombinant P2X(2) receptors. Wild type and 31 mutant P2X(2) receptors were expressed in HEK-293 cells and studied under voltage-clamp. We found that the relative Ca(2+) permeability measured from the reversal potentials of ATP-gated currents was unaffected by neutralizing fixed charge (Asp(315), Asp(349)) near the mouths of the channel pore. By contrast, mutations that changed the character or side chain volume of three polar residues (Thr(336), Thr(339), Ser(340)) within the pore led to significant changes in P(Ca)/P(Cs). The largest changes occurred when Thr(339) and Ser(340) were replaced with tyrosine; these mutations almost completely abolished Ca(2+) permeability, reduced P(Li)/P(Cs) by about one-half, and shifted the relative permeability sequence of Cs(+), Rb(+), K(+), and Na(+) to their relative mobility in water. Our results suggest that the permeability sequence of the P2X(2) receptor arises in part from interactions of permeating cations with the polar side chains of three amino acids located in a short stretch of the second transmembrane domain. PMID- 11402045 TI - Recombinant forms of tetanus toxin engineered for examining and exploiting neuronal trafficking pathways. AB - Tetanus toxin is a fascinating, multifunctional protein that binds to peripheral neurons, undergoes retrograde transport and trans-synaptic transfer to central inhibitory neurons where it blocks transmitter release, thereby, causing spastic paralysis. As a pre-requisite for exploiting its unique trafficking properties, a novel recombinant single chain was expressed at a high level in Escherichia coli as a soluble, easily purifiable protein. It could be activated with enterokinase to produce a dichain that matched native toxin in terms of proteolytic and neuroinhibitory activities, as well as induction of spastic paralysis in mice. Importantly, nicking was not essential for protease activity. Substitution of Glu(234) by Ala created a protease-deficient atoxic form, which blocked the neuroparalytic action of tetanus toxin in vitro, with equal potency to its heavy chain; but, the mutant proved >30-fold more potent in preventing tetanus in mice. This observation unveils differences between the intoxication processes resulting from retrograde transport of toxin in vivo and its local uptake into peripheral or central nerves in vitro, dispelling a popularly held belief that the heavy chain is the sole determinant for efficient trafficking. Thus, this innocuous mutant may be a useful vehicle, superior to the heavy chain, for drug delivery to central neurons. PMID- 11402046 TI - Intracellular Ca2+ and Zn2+ levels regulate the alternative cell density dependent secretion of S100B in human glioblastoma cells. AB - In recent years, protein translocation has been implicated as the mechanism that controls assembly of signaling complexes and induction of signaling cascades. Several members of the multifunctional Ca(2+)- (Zn(2+)- and Cu(2+))-binding S100 proteins appear to translocate upon cellular stimulation, and some are even secreted from cells, exerting extracellular functions. We transfected cells with S100B-green fluorescent fusion proteins and followed the relocation in real time. A small number of cells underwent translocation spontaneously. However, the addition of thapsigargin, which increases Ca(2+) levels, intensified ongoing translocation and secretion or induced these processes in resting cells. On the other hand, EGTA or BAPTA (1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid), the Ca(2+)-chelating agents, inhibited these processes. In contrast, relocation of S100B seemed to be negatively dependent on Zn(2+) levels. Treatment of cells with TPEN (N,N,N',N'-tetrakis(2-pyridylmethyl)ethylenediamine), a Zn(2+) binding drug, resulted in a dramatic redistribution and translocation of S100B. Secretion of S100B, when measured by ELISA, was dependent on cell density. As cells reached confluence the secretion drastically declined. However, an increase in Ca(2+) levels, and even more so, a decrease in Zn(2+) concentration, reactivated secretion of S100B. On the other hand, secretion did not decrease by treatment with brefeldin A, supporting the view that this process is independent of the endoplasmic reticulum-Golgi classical secretion pathway. PMID- 11402047 TI - A dual signaling cascade that regulates the ectodomain shedding of heparin binding epidermal growth factor-like growth factor. AB - Ectodomain shedding is an important mechanism to regulate the biological activities of membrane proteins. We focus here on the signaling mechanism of the ectodomain shedding of heparin-binding epidermal growth factor (EGF)-like growth factor (pro HB-EGF). Lysophosphatidic acid (LPA), a ligand for seven transmembrane G protein-coupled receptors, stimulates the shedding of pro HB-EGF, which constitutes a G protein-coupled receptor-mediated transactivation of the EGF receptor. Experiments using a series of inhibitors and overexpression of mutant forms of signaling molecules revealed that the Ras-Raf-MEK signal is essential for the LPA-induced shedding. In addition, the small GTPase Rac is involved in the LPA-induced shedding, possibly to promote MEK activation. 12-O Tetradecanoylphorbol-13-acetate is another potent inducer of pro HB-EGF shedding. We also demonstrate that the LPA-induced pathway is distinct from the 12-O tetradecanoylphorbol-13-acetate-induced pathway and that these pathways constitute a dual signaling cascade that regulates the shedding of pro HB-EGF. PMID- 11402048 TI - Insulin-stimulated activation of eNOS is independent of Ca2+ but requires phosphorylation by Akt at Ser(1179). AB - Vasodilator actions of insulin are mediated by activation of endothelial nitric oxide synthase (eNOS) and subsequent production of NO. Phosphatidylinositol 3 kinase and Akt play important roles in insulin-signaling pathways leading to production of NO in vascular endothelium. Here we dissected mechanisms whereby insulin activates eNOS by using the fluorescent dye DAF-2 to directly measure NO production in single cells. Insulin caused a rapid increase in intracellular NO in NIH-3T3(IR) cells transiently transfected with eNOS. The stimulation of NO production by lysophosphatidic acid (LPA) was abrogated by pretreatment of cells with the calcium chelator 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid. Remarkably, in the same cells, insulin-stimulated production of NO was unaffected. However, cells expressing the eNOS-S1179A mutant (disrupted Akt phosphorylation site) did not produce detectable NO in response to insulin, whereas the response to LPA was similar to that observed in cells expressing wild type eNOS. Moreover, production of NO in response to insulin was blocked by coexpression of an inhibitory mutant of Akt, whereas the response to LPA was unaffected. Phosphorylation of eNOS at Ser(1179) was observed only in response to treatment with insulin, but not with LPA. Interestingly, platelet-derived growth factor treatment of cells activated Akt but not eNOS. Results from human vascular endothelial cells were qualitatively similar to those obtained in transfected NIH 3T3(IR) cells, although the magnitude of the responses was smaller. We conclude that insulin regulates eNOS activity using a Ca(2+)-independent mechanism requiring phosphorylation of eNOS by Akt. Importantly, phosphorylation-dependent mechanisms that enhance eNOS activity can operate independently from Ca(2+) dependent mechanisms. PMID- 11402049 TI - Mechanisms of pH regulation in the regulated secretory pathway. AB - A precise pH gradient between organelles of the regulated secretory pathway is required for sorting and processing of prohormones. We studied pH regulation in live endocrine cells by targeting biotin-based pH indicators to cellular organelles expressing avidin-chimera proteins. In AtT-20 cells, we found that steady-state pH decreased from the endoplasmic reticulum (ER) (pH(ER) = 7.4 +/- 0.2, mean +/- S.D.) to Golgi (pH(G) = 6.2 +/- 0.4) to mature secretory granules (MSGs) (pH(MSG) = 5.5 +/- 0.4). Golgi and MSGs required active H(+) v-ATPases for acidification. ER, Golgi, and MSG steady-state pH values were also dependent upon the different H(+) leak rates across each membrane. However, neither steady-state pH(MSG) nor rates of passive H(+) leak were affected by Cl(-)-free solutions or valinomycin, indicating that MSG membrane potential was small and not a determinant of pH(MSG). Therefore, our data do not support earlier suggestions that organelle acidification is primarily regulated by Cl(-) conductances. Measurements of H(+) leak rates, buffer capacities, and estimates of surface areas and volumes of these organelles were applied to a mathematical model to determine the H(+) permeability (P(H+)) of each organelle membrane. We found that P(H+) decreased progressively from ER to Golgi to MSGs, and proper acidification of Golgi and MSGs required gradual decreases in P(H+) and successive increases in the active H(+) pump density. PMID- 11402050 TI - Crystal structure of baculovirus P35 reveals a novel conformational change in the reactive site loop after caspase cleavage. AB - Baculovirus P35 is a universal suppressor of apoptosis that stoichiometrically inhibits cellular caspases in a novel cleavage-dependent mechanism. Upon caspase cleavage at Asp-87, the 10- and 25-kDa cleavage products of P35 remain tightly associated with the inhibited caspase. Mutations in the alpha-helix of the reactive site loop preceding the cleavage site abrogate caspase inhibition and antiapoptotic activity. Substitution of Pro for Val-71, which is located in the middle of this alpha-helix, produces a protein that is cleaved at the requisite Asp-87 but does not remain bound to the caspase. This loss-of-function mutation provided the opportunity to structurally analyze the conformational changes of the P35 reactive site loop after caspase cleavage. We report here the 2.7 A resolution crystal structure of V71P-mutated P35 after cleavage by human caspase 3. The structure reveals a large movement in the carboxyl-terminal side of the reactive site loop that swings down and forms a new beta-strand that augments an existing beta-sheet. Additionally, the hydrophobic amino terminus releases and extends away from the protein core. Similar movements occur when P35 forms an inhibitory complex with human caspase-8. These findings suggest that the alpha helix mutation may alter the sequential steps or kinetics of the conformational changes required for inhibition, thereby causing P35 loss of function. PMID- 11402051 TI - Identification of a putative sordarin binding site in Candida albicans elongation factor 2 by photoaffinity labeling. AB - Candida albicans EF-2 binds sordarin to a single class of binding sites with K(d) = 1.26 microm. Equimolar mixtures of EF-2 and ribosomes, in the presence of a non hydrolyzable GTP analog, reveal two classes of high affinity sordarin binding sites with K(d) = 0.7 and 41.5 nm, probably due to the existence of two ribosome populations. Photoaffinity labeling of C. albicans EF-2 in the absence of ribosomes has been performed with [(14)C]GM258383, a photoactivatable sordarin derivative. Labeling is saturable and can be considered specific, because it can be prevented with another sordarin analog. The fragment Gln(224)-Lys(232) has been identified as the modified peptide within the EF-2 sequence, Lys(228) being the residue to which the photoprobe was linked. This fragment is included within the G"-subdomain of EF-2. These results are discussed in the light of the high sordarin specificity toward fungal systems. PMID- 11402052 TI - Identification of inactivation determinants in the domain IIS6 region of high voltage-activated calcium channels. AB - We have recently reported that transfer of the domain IIS6 region from rapidly inactivating R-type (alpha(1E)) calcium channels to slowly inactivating L-type (alpha(1C)) calcium channel confers rapid inactivation (Stotz, S. C., Hamid, J., Spaetgens, R. L., Jarvis, S. E., and Zamponi, G. W. (2000) J. Biol. Chem. 275, 24575-24582). Here we have identified individual amino acid residues in the IIS6 regions that are responsible for these effects. In this region, alpha(1C) and alpha(1E) channels differ in seven residues, and exchanging five of those residues individually or in combination did not significantly affect inactivation kinetics. By contrast, replacement of residues Phe-823 or Ile-829 of alpha(1C) with the corresponding alpha(1E) residues significantly accelerated inactivation rates and, when substituted concomitantly, approached the rapid inactivation kinetics of R-type channels. A systematic substitution of these residues with a series of other amino acids revealed that decreasing side chain size at position 823 accelerates inactivation, whereas a dependence of the inactivation kinetics on the degree of hydrophobicity could be observed at position 829. Although these point mutations facilitated rapid entry into the inactivated state of the channel, they had little to no effect on the rate of recovery from inactivation. This suggests that the development of and recovery from inactivation are governed by separate structural determinants. Finally, the effects of mutations that accelerated alpha(1C) inactivation could still be antagonized following coexpression of the rat beta(2a) subunit or by domain I-II linker substitutions that produce ultra slow inactivation of wild type channels, indicating that the inactivation kinetics seen with the mutants remain subject to regulation by the domain I-II linker. Overall, our results provide novel insights into a complex process underlying calcium channel inactivation. PMID- 11402053 TI - Amino acid determinants in cyclooxygenase-2 oxygenation of the endocannabinoid 2 arachidonylglycerol. AB - The endocannabinoid, 2-arachidonylglycerol (2-AG), is an endogenous ligand for the central (CB1) and peripheral (CB2) cannabinoid receptors and has been shown to be efficiently and selectively oxygenated by cyclooxygenase (COX)-2. We have investigated 2-AG/COX-2 interactions through site-directed mutagenesis. An evaluation of more than 20 site-directed mutants of murine COX-2 has allowed for the development of a model of 2-AG binding within the COX-2 active site. Most strikingly, these studies have identified Arg-513 as a critical determinant in the ability of COX-2 to efficiently generate prostaglandin H(2) glycerol ester, explaining, in part, the observed isoform selectivity for this substrate. Mutational analysis of Leu-531, an amino acid located directly across from Arg 513 in the COX-2 active site, suggests that 2-AG is shifted in the active site away from this hydrophobic residue and toward Arg-513 relative to arachidonic acid. Despite this difference, aspirin-treated COX-2 oxygenates 2-AG to afford 15 hydroxyeicosatetraenoic acid glycerol ester in a reaction analogous to the C-15 oxygenation of arachidonic acid observed with acetylated COX-2. Finally, the differences in substrate binding do not alter the stereospecificity of the cyclooxygenase reaction; 2-AG-derived and arachidonic acid-derived products share identical stereochemistry. PMID- 11402054 TI - Dual role of inflammatory stimuli in activation-induced cell death of mouse microglial cells. Initiation of two separate apoptotic pathways via induction of interferon regulatory factor-1 and caspase-11. AB - We have previously shown that mouse microglial cells undergo apoptosis upon inflammatory activation and that nitric oxide (NO) is the major autocrine mediator in this process (Lee, P., Lee, J., Kim, S., Yagita, H., Lee, M. S., Kim, S. Y., Kim, H., and Suk, K. (2001) Brain Res. 892, 380-385). Here, we present evidence that interferon regulatory factor-1 (IRF-1) and caspase-11 are the essential molecules in activation-induced cell death of microglial cells. The apoptogenic action of inflammatory stimuli such as lipopolysaccharide (LPS) and interferon-gamma (IFNgamma) was mediated through the induction of IRF-1 and caspase-11 expression in two separate events. Although IRF-1 was required for NO synthesis, caspase-11 induction was necessary for NO-independent apoptotic pathway. Microglial cells from IRF-1-deficient mice showed markedly decreased NO production, and they were partially resistant to apoptosis in response to LPS/IFNgamma but were sensitive to NO donor exposure. LPS/IFNgamma treatment resulted in the induction of caspase-11 followed by activation of caspase-11, -1, and -3. Inactivation of caspase-11 by the transfection of dominant-negative mutant or treatment with the caspase inhibitors rendered microglial cells partially resistant to LPS/IFNgamma-induced apoptosis. Inhibition of both NO synthesis and caspase-11 completely blocked LPS/IFNgamma-induced cytotoxicity. These results indicated that LPS/IFNgamma not only induced the production of cytotoxic NO through IRF-1 but also initiated the NO-independent apoptotic pathway through the induction of caspase-11 expression. PMID- 11402055 TI - Disassociation of MAPK activation and c-Fos expression in F9 embryonic carcinoma cells following retinoic acid-induced endoderm differentiation. AB - Retinoic acid induces cell differentiation and suppresses cell growth in a wide spectrum of cell lines, and down-regulation of activator protein-1 activity by retinoic acid contributes to these effects. In embryonic stem cell-like F9 teratocarcinoma cells, which are widely used to study retinoic acid actions on gene regulation and early embryonic differentiation, retinoic acid treatment for 4 days resulted in suppression of cell growth and differentiation into primitive and then visceral endoderm-like cells, accompanied by a suppression of serum induced c-Fos expression. The MAPK (ERK) pathway was involved in mitogenic signaling in F9 cells stimulated with serum. Surprisingly, although c-Fos expression was reduced, the MAPK activity was not decreased by retinoic acid treatment. We found that retinoic acid treatment inhibited the phosphorylation of Elk-1, a target of activated MAPK required for c-Fos transcription. In F9 cells, the MAPK/MEK inhibitor PD98059 suppressed Elk-1 phosphorylation and c-Fos expression, indicating that MAPK activity is required for Elk-1 phosphorylation/activation. Phosphoprotein phosphatase 2B (calcineurin), the major phosphatase for activated Elk-1, is not the target in the disassociation of MAPK activation and c-Fos expression since its inhibition by cyclosporin A or activation by ionomycin had no significant effects on serum-stimulated c-Fos expression and Elk-1 phosphorylation. Thus, we conclude that retinoic acid treatment to induce F9 cell differentiation uncouples Ras/MAPK activation from c Fos expression by reduction of Elk-1 phosphorylation through a mechanism not involving the activation of phosphoprotein phosphatase 2B. PMID- 11402056 TI - The stability of the TFIIA-TBP-DNA complex is dependent on the sequence of the TATAAA element. AB - To determine the mechanistic differences between canonical and noncanonical TATA elements, we compared the functional activity of two sequences: TATAAA (canonical) and CATAAA (noncanonical). The TATAAA element can support high levels of transcription in vivo, whereas the CATAAA element is severely defective for this function. This dramatic functional difference is not likely to be due to a difference in TBP (TATA-binding protein) binding efficiency because protein-DNA complex studies in vitro indicate little difference between the two DNA sequences in the formation and stability of the TBP-DNA complex. In addition, the binding and stability of the TFIIB-TBP-DNA complex is similar for the two elements. In striking contrast, the TFIIA-TBP-DNA complex is significantly less stable on the CATAAA element when compared with the TATAAA element. A role for TFIIA in distinguishing between TATAAA and CATAAA in vivo was tested by fusing a subunit of TFIIA to TBP. We found that fusion of TFIIA to TBP dramatically increases transcription from CATAAA in yeast cells. Taken together, these results indicate that the stability of the TFIIA-TBP complex depends strongly on the sequence of the core promoter element and that the TFIIA-TBP complex plays an important function in recognizing optimal promoters in vivo. PMID- 11402057 TI - Electron microscopy of cells: a new beginning for a new century. PMID- 11402059 TI - Multiple distinct targeting signals in integral peroxisomal membrane proteins. AB - Peroxisomal proteins are synthesized on free polysomes and then transported from the cytoplasm to peroxisomes. This process is mediated by two short well-defined targeting signals in peroxisomal matrix proteins, but a well-defined targeting signal has not yet been described for peroxisomal membrane proteins (PMPs). One assumption in virtually all prior studies of PMP targeting is that a given protein contains one, and only one, distinct targeting signal. Here, we show that the metabolite transporter PMP34, an integral PMP, contains at least two nonoverlapping sets of targeting information, either of which is sufficient for insertion into the peroxisome membrane. We also show that another integral PMP, the peroxin PEX13, also contains two independent sets of peroxisomal targeting information. These results challenge a major assumption of most PMP targeting studies. In addition, we demonstrate that PEX19, a factor required for peroxisomal membrane biogenesis, interacts with the two minimal targeting regions of PMP34. Together, these results raise the interesting possibility that PMP import may require novel mechanisms to ensure the solubility of integral PMPs before their insertion in the peroxisome membrane, and that PEX19 may play a central role in this process. PMID- 11402060 TI - Tom40, the pore-forming component of the protein-conducting TOM channel in the outer membrane of mitochondria. AB - Tom40 is the main component of the preprotein translocase of the outer membrane of mitochondria (TOM complex). We have isolated Tom40 of Neurospora crassa by removing the receptor Tom22 and the small Tom components Tom6 and Tom7 from the purified TOM core complex. Tom40 is organized in a high molecular mass complex of approximately 350 kD. It forms a high conductance channel. Mitochondrial presequence peptides interact specifically with Tom40 reconstituted into planar lipid membranes and decrease the ion flow through the pores in a voltage dependent manner. The secondary structure of Tom40 comprises approximately 31% beta-sheet, 22% alpha-helix, and 47% remaining structure as determined by circular dichroism measurements and Fourier transform infrared spectroscopy. Electron microscopy of purified Tom40 revealed particles primarily with one center of stain accumulation. They presumably represent an open pore with a diameter of approximately 2.5 nm, similar to the pores found in the TOM complex. Thus, Tom40 is the core element of the TOM translocase; it forms the protein conducting channel in an oligomeric assembly. PMID- 11402061 TI - Tumor suppressor PTEN inhibits nuclear accumulation of beta-catenin and T cell/lymphoid enhancer factor 1-mediated transcriptional activation. AB - beta-Catenin is a protein that plays a role in intercellular adhesion as well as in the regulation of gene expression. The latter role of beta-catenin is associated with its oncogenic properties due to the loss of expression or inactivation of the tumor suppressor adenomatous polyposis coli (APC) or mutations in beta-catenin itself. We now demonstrate that another tumor suppressor, PTEN, is also involved in the regulation of nuclear beta-catenin accumulation and T cell factor (TCF) transcriptional activation in an APC independent manner. We show that nuclear beta-catenin expression is constitutively elevated in PTEN null cells and this elevated expression is reduced upon reexpression of PTEN. TCF promoter/luciferase reporter assays and gel mobility shift analysis demonstrate that PTEN also suppresses TCF transcriptional activity. Furthermore, the constitutively elevated expression of cyclin D1, a beta-catenin/TCF-regulated gene, is also suppressed upon reexpression of PTEN. Mechanistically, PTEN increases the phosphorylation of beta catenin and enhances its rate of degradation. We define a pathway that involves mainly integrin-linked kinase and glycogen synthase kinase 3 in the PTEN dependent regulation of beta-catenin stability, nuclear beta-catenin expression, and transcriptional activity. Our data indicate that beta-catenin/TCF-mediated gene transcription is regulated by PTEN, and this may represent a key mechanism by which PTEN suppresses tumor progression. PMID- 11402062 TI - Focal contacts as mechanosensors: externally applied local mechanical force induces growth of focal contacts by an mDia1-dependent and ROCK-independent mechanism. AB - The transition of cell-matrix adhesions from the initial punctate focal complexes into the mature elongated form, known as focal contacts, requires GTPase Rho activity. In particular, activation of myosin II-driven contractility by a Rho target known as Rho-associated kinase (ROCK) was shown to be essential for focal contact formation. To dissect the mechanism of Rho-dependent induction of focal contacts and to elucidate the role of cell contractility, we applied mechanical force to vinculin-containing dot-like adhesions at the cell edge using a micropipette. Local centripetal pulling led to local assembly and elongation of these structures and to their development into streak-like focal contacts, as revealed by the dynamics of green fluorescent protein-tagged vinculin or paxillin and interference reflection microscopy. Inhibition of Rho activity by C3 transferase suppressed this force-induced focal contact formation. However, constitutively active mutants of another Rho target, the formin homology protein mDia1 (Watanabe, N., T. Kato, A. Fujita, T. Ishizaki, and S. Narumiya. 1999. Nat. Cell Biol. 1:136-143), were sufficient to restore force-induced focal contact formation in C3 transferase-treated cells. Force-induced formation of the focal contacts still occurred in cells subjected to myosin II and ROCK inhibition. Thus, as long as mDia1 is active, external tension force bypasses the requirement for ROCK-mediated myosin II contractility in the induction of focal contacts. Our experiments show that integrin-containing focal complexes behave as individual mechanosensors exhibiting directional assembly in response to local force. PMID- 11402063 TI - Intracellular retention of newly synthesized insulin in yeast is caused by endoproteolytic processing in the Golgi complex. AB - An insulin-containing fusion protein (ICFP, encoding the yeast prepro-alpha factor leader peptide fused via a lysine-arginine cleavage site to a single chain insulin) has been expressed in Saccharomyces cerevisiae where it is inefficiently secreted. Single gene disruptions have been identified that cause enhanced immunoreactive insulin secretion (eis). Five out of six eis mutants prove to be vacuolar protein sorting (vps)8, vps35, vps13, vps4, and vps36, which affect Golgi<-->endosome trafficking. Indeed, in wild-type yeast insulin is ultimately delivered to the vacuole, whereas vps mutants secrete primarily unprocessed ICFP. Disruption of KEX2, which blocks intracellular processing to insulin, quantitatively reroutes ICFP to the cell surface, whereas loss of the Vps10p sorting receptor is without effect. Secretion of unprocessed ICFP is not based on a dominant secretion signal in the alpha-leader peptide. Although insulin sorting mediated by Kex2p is saturable, Kex2p functions not as a sorting receptor but as a protease: replacement of Kex2p by truncated secretory Kex2p (which travels from Golgi to cell surface) still causes endoproteolytic processing and intracellular insulin retention. Endoproteolysis promotes a change in insulin's biophysical properties. B5His residues normally participate in multimeric insulin packing; a point mutation at this position permits ICFP processing but causes the majority of processed insulin to be secreted. The data argue that multimeric assembly consequent to endoproteolytic maturation regulates insulin sorting in the secretory pathway. PMID- 11402064 TI - HCP-4, a CENP-C-like protein in Caenorhabditis elegans, is required for resolution of sister centromeres. AB - The centromere plays a critical role in the segregation of chromosomes during mitosis. In mammals, sister centromeres are resolved from one another in the G2 phase of the cell cycle. During prophase, chromosomes condense with sister centromeres oriented in a back to back configuration enabling only one chromatid to be captured by each half spindle. To study this process, we identified a centromere protein (CENP)-C-like protein, holocentric protein (HCP)-4, in Caenorhabditis elegans based on sequence identity, loss of function phenotype, and centromeric localization. HCP-4 is found in the cytoplasm during interphase, but is nuclear localized in mitosis, where it localizes specifically to the centromere. The localization of HCP-4 to the centromere is dependent on the centromeric histone HCP-3; in addition, HCP-3 and HCP-4 are both required for localization of a CENP-F-like protein, HCP-1, indicating an ordered assembly pathway. Loss of HCP-4 expression by RNA-mediated interference resulted in a failure to generate resolution of sister centromeres on chromosomes, suggesting that HCP-4 is required for sister centromere resolution. These chromosomes also failed to form a functional kinetochore. Thus, the CENP-C-like protein HCP-4 is essential for both resolution sister centromeres and attachment to the mitotic spindle. PMID- 11402065 TI - Functional analysis of kinetochore assembly in Caenorhabditis elegans. AB - In all eukaryotes, segregation of mitotic chromosomes requires their interaction with spindle microtubules. To dissect this interaction, we use live and fixed assays in the one-cell stage Caenorhabditis elegans embryo. We compare the consequences of depleting homologues of the centromeric histone CENP-A, the kinetochore structural component CENP-C, and the chromosomal passenger protein INCENP. Depletion of either CeCENP-A or CeCENP-C results in an identical "kinetochore null" phenotype, characterized by complete failure of mitotic chromosome segregation as well as failure to recruit other kinetochore components and to assemble a mechanically stable spindle. The similarity of their depletion phenotypes, combined with a requirement for CeCENP-A to localize CeCENP-C but not vice versa, suggest that a key step in kinetochore assembly is the recruitment of CENP-C by CENP-A-containing chromatin. Parallel analysis of CeINCENP-depleted embryos revealed mitotic chromosome segregation defects different from those observed in the absence of CeCENP-A/C. Defects are observed before and during anaphase, but the chromatin separates into two equivalently sized masses. Mechanically stable spindles assemble that show defects later in anaphase and telophase. Furthermore, kinetochore assembly and the recruitment of CeINCENP to chromosomes are independent. These results suggest distinct roles for the kinetochore and the chromosomal passengers in mitotic chromosome segregation. PMID- 11402066 TI - HIM-10 is required for kinetochore structure and function on Caenorhabditis elegans holocentric chromosomes. AB - Macromolecular structures called kinetochores attach and move chromosomes within the spindle during chromosome segregation. Using electron microscopy, we identified a structure on the holocentric mitotic and meiotic chromosomes of Caenorhabditis elegans that resembles the mammalian kinetochore. This structure faces the poles on mitotic chromosomes but encircles meiotic chromosomes. Worm kinetochores require the evolutionarily conserved HIM-10 protein for their structure and function. HIM-10 localizes to the kinetochores and mediates attachment of chromosomes to the spindle. Depletion of HIM-10 disrupts kinetochore structure, causes a failure of bipolar spindle attachment, and results in chromosome nondisjunction. HIM-10 is related to the Nuf2 kinetochore proteins conserved from yeast to humans. Thus, the extended kinetochores characteristic of C. elegans holocentric chromosomes provide a guide to the structure, molecular architecture, and function of conventional kinetochores. PMID- 11402067 TI - Spindle checkpoint protein Bub1 is required for kinetochore localization of Mad1, Mad2, Bub3, and CENP-E, independently of its kinase activity. AB - The spindle checkpoint inhibits the metaphase to anaphase transition until all the chromosomes are properly attached to the mitotic spindle. We have isolated a Xenopus homologue of the spindle checkpoint component Bub1, and investigated its role in the spindle checkpoint in Xenopus egg extracts. Antibodies raised against Bub1 recognize a 150-kD phosphoprotein at both interphase and mitosis, but the molecular mass is reduced to 140 upon dephosphorylation in vitro. Bub1 is essential for the establishment and maintenance of the checkpoint and is localized to kinetochores, similar to the spindle checkpoint complex Mad1-Mad2. However, Bub1 differs from Mad1-Mad2 in that Bub1 remains on kinetochores that have attached to microtubules; the protein eventually dissociates from the kinetochore during anaphase. Immunodepletion of Bub1 abolishes the spindle checkpoint and the kinetochore binding of the checkpoint proteins Mad1, Mad2, Bub3, and CENP-E. Interestingly, reintroducing either wild-type or kinase deficient Bub1 protein restores the checkpoint and the kinetochore localization of these proteins. Our studies demonstrate that Bub1 plays a central role in triggering the spindle checkpoint signal from the kinetochore, and that its kinase activity is not necessary for the spindle checkpoint in Xenopus egg extracts. PMID- 11402068 TI - A novel integrin-linked kinase-binding protein, affixin, is involved in the early stage of cell-substrate interaction. AB - Focal adhesions (FAs) are essential structures for cell adhesion, migration, and morphogenesis. Integrin-linked kinase (ILK), which is capable of interacting with the cytoplasmic domain of beta1 integrin, seems to be a key component of FAs, but its exact role in cell-substrate interaction remains to be clarified. Here, we identified a novel ILK-binding protein, affixin, that consists of two tandem calponin homology domains. In CHOcells, affixin and ILK colocalize at FAs and at the tip of the leading edge, whereas in skeletal muscle cells they colocalize at the sarcolemma where cells attach to the basal lamina, showing a striped pattern corresponding to cytoplasmic Z-band striation. When CHO cells are replated on fibronectin, affixin and ILK but not FA kinase and vinculin concentrate at the cell surface in blebs during the early stages of cell spreading, which will grow into membrane ruffles on lamellipodia. Overexpression of the COOH-terminal region of affixin, which is phosphorylated by ILK in vitro, blocks cell spreading at the initial stage, presumably by interfering with the formation of FAs and stress fibers. The coexpression of ILK enhances this effect. These results provide evidence suggesting that affixin is involved in integrin-ILK signaling required for the establishment of cell-substrate adhesion. PMID- 11402069 TI - Bax and Bak coalesce into novel mitochondria-associated clusters during apoptosis. AB - Bax is a member of the Bcl-2 family of proteins known to regulate mitochondria dependent programmed cell death. Early in apoptosis, Bax translocates from the cytosol to the mitochondrial membrane. We have identified by confocal and electron microscopy a novel step in the Bax proapoptotic mechanism immediately subsequent to mitochondrial translocation. Bax leaves the mitochondrial membranes and coalesces into large clusters containing thousands of Bax molecules that remain adjacent to mitochondria. Bak, a close homologue of Bax, colocalizes in these apoptotic clusters in contrast to other family members, Bid and Bad, which circumscribe the outer mitochondrial membrane throughout cell death progression. We found the formation of Bax and Bak apoptotic clusters to be caspase independent and inhibited completely and specifically by Bcl-X(L), correlating cluster formation with cytotoxic activity. Our results reveal the importance of a novel structure formed by certain Bcl-2 family members during the process of cell death. PMID- 11402070 TI - CD44 is a major E-selectin ligand on human hematopoietic progenitor cells. AB - E-selectin plays a critical role in mediating tissue-specific homing of T cells into skin, and of primitive hematopoietic progenitor cells (HPCs) into bone marrow (BM). Though it is known that a glycoform of PSGL-1 (CLA) functions as the principal E-selectin ligand on human T lymphocytes, the E-selectin ligand(s) of human HPCs has not been identified. We used a shear-based adherence assay to analyze and define the E-selectin ligand activity of membrane proteins from human HPCs. Our data show that PSGL-1 expressed on human HPCs is an E-selectin ligand, and that HPCs also express a previously unrecognized E-selectin ligand, CD44. The E-selectin ligand activity of CD44 is conferred by the elaboration of sialylated, fucosylated binding determinants on N-glycans. This glycoform of CD44 is expressed on primitive CD34+ human HPCs, but not on more mature hematopoietic cells. Under physiologic flow conditions, this molecule mediates E-selectin dependent rolling interactions over a wider shear range than that of PSGL-1, and promotes human HPC rolling interactions on E-selectin expressed on human BM endothelial cells. These findings offer new insights into the structural biology and physiology of CD44, and into the molecular basis of E-selectin-dependent adhesive interactions that direct homing of human HPC to BM. PMID- 11402071 TI - Subdomain-specific localization of CLIMP-63 (p63) in the endoplasmic reticulum is mediated by its luminal alpha-helical segment. AB - The microtubule-binding integral 63 kD cytoskeleton-linking membrane protein (CLIMP-63; former name, p63) of the rough endoplasmic reticulum (ER) is excluded from the nuclear envelope. We studied the mechanism underlying this ER subdomain specific localization by mutagenesis and structural analysis. Deleting the luminal but not cytosolic segment of CLIMP-63 abrogated subdomain-specific localization, as visualized by confocal microscopy in living cells and by immunoelectron microscopy using ultrathin cryosections. Photobleaching/recovery analysis revealed that the luminal segment determines restricted diffusion and immobility of the protein. The recombinant full-length luminal segment of CLIMP 63 formed alpha-helical 91-nm long rod-like structures as evident by circular dichroism spectroscopy and electron microscopy. In the analytical ultracentrifuge, the luminal segment sedimented at 25.7 S, indicating large complexes. The complexes most likely arose by electrostatic interactions of individual highly charged coiled coils. The findings indicate that the luminal segment of CLIMP-63 is necessary and sufficient for oligomerization into alpha helical complexes that prevent nuclear envelope localization. Concentration of CLIMP-63 into patches may enhance microtubule binding on the cytosolic side and contribute to ER morphology by the formation of a protein scaffold in the lumen of the ER. PMID- 11402072 TI - SERCA1 truncated proteins unable to pump calcium reduce the endoplasmic reticulum calcium concentration and induce apoptosis. AB - By pumping calcium from the cytosol to the ER, sarco/endoplasmic reticulum calcium ATPases (SERCAs) play a major role in the control of calcium signaling. We describe two SERCA1 splice variants (S1Ts) characterized by exon 4 and/or exon 11 splicing, encoding COOH terminally truncated proteins, having only one of the seven calcium-binding residues, and thus unable to pump calcium. As shown by semiquantitative RT-PCR, S1T transcripts are differentially expressed in several adult and fetal human tissues, but not in skeletal muscle and heart. S1T proteins expression was detected by Western blot in nontransfected cell lines. In transiently transfected cells, S1T homodimers were revealed by Western blot using mildly denaturing conditions. S1T proteins were shown, by confocal scanning microscopy, to colocalize with endogenous SERCA2b into the ER membrane. Using ER targeted aequorin (erAEQ), we have found that S1T proteins reduce ER calcium and reverse elevation of ER calcium loading induced by SERCA1 and SERCA2b. Our results also show that SERCA1 variants increase ER calcium leakage and are consistent with the hypothesis of a cation channel formed by S1T homodimers. Finally, when overexpressed in liver-derived cells, S1T proteins significantly induce apoptosis. These data reveal a further mechanism modulating Ca(2+) accumulation into the ER of nonmuscle cells and highlight the relevance of S1T proteins to the control of apoptosis. PMID- 11402073 TI - Localization of calmodulin and dynein light chain LC8 in flagellar radial spokes. AB - Genetic and in vitro analyses have revealed that radial spokes play a crucial role in regulation of ciliary and flagellar motility, including control of waveform. However, the mechanisms of regulation are not understood. Here, we developed a novel procedure to isolate intact radial spokes as a step toward understanding the mechanism by which these complexes regulate dynein activity. The isolated radial spokes sediment as 20S complexes that are the size and shape of radial spokes. Extracted radial spokes rescue radial spoke structure when reconstituted with isolated axonemes derived from the radial spoke mutant pf14. Isolated radial spokes are composed of the 17 previously defined spoke proteins as well as at least five additional proteins including calmodulin and the ubiquitous dynein light chain LC8. Analyses of flagellar mutants and chemical cross-linking studies demonstrated calmodulin and LC8 form a complex located in the radial spoke stalk. We postulate that calmodulin, located in the radial spoke stalk, plays a role in calcium control of flagellar bending. PMID- 11402074 TI - Prion filament networks in [URE3] cells of Saccharomyces cerevisiae. AB - The [URE3] prion (infectious protein) of yeast is a self-propagating, altered form of Ure2p that cannot carry out its normal function in nitrogen regulation. Previous data have shown that Ure2p can form protease-resistant amyloid filaments in vitro, and that it is aggregated in cells carrying the [URE3] prion. Here we show by electron microscopy that [URE3] cells overexpressing Ure2p contain distinctive, filamentous networks in their cytoplasm, and demonstrate by immunolabeling that these networks contain Ure2p. In contrast, overexpressing wild-type cells show a variety of Ure2p distributions: usually, the protein is dispersed sparsely throughout the cytoplasm, although occasionally it is found in multiple small, focal aggregates. However, these distributions do not resemble the single, large networks seen in [URE3] cells, nor do the control cells exhibit cytoplasmic filaments. In [URE3] cell extracts, Ure2p is present in aggregates that are only partially solubilized by boiling in SDS and urea. In these aggregates, the NH(2)-terminal prion domain is inaccessible to antibodies, whereas the COOH-terminal nitrogen regulation domain is accessible. This finding is consistent with the proposal that the prion domains stack to form the filament backbone, which is surrounded by the COOH-terminal domains. These observations support and further specify the concept of the [URE3] prion as a self-propagating amyloid. PMID- 11402076 TI - Here, there, and everywhere: kinetochore function on holocentric chromosomes. PMID- 11402077 TI - Early improvement during manual-guided cognitive and dynamic psychotherapies predicts 16-week remission status. AB - This study examined the extent to which improvement from baseline to weeks 2, 3, and 4 on the Beck Depression Inventory and Beck Anxiety Inventory predict week 16 clinical remission for patients with major depressive disorder, generalized anxiety disorder, and/or obsessive-compulsive or avoidant personality disorders who were receiving manual-based psychotherapies. Logistic regression and receiver operator characteristic analyses revealed relatively accurate identification of remitters and nonremitters based on improvement from baseline to sessions 2 to 4 in both original and cross-validation samples. Predictive success did not vary as a function of diagnosis, treatment type (cognitive or dynamic), or treatment status (short-term or long-term). The clinical implications of the results are discussed. PMID- 11402078 TI - Contribution of patient defense mechanisms and therapist interventions to the development of early therapeutic alliance in a brief psychodynamic investigation. AB - This preliminary study examined how patients' defense mechanisms and psychotherapists' techniques influence early alliance formation. The authors assessed the relationships among defense mechanisms, therapist interventions, and the development of alliance in a sample of 12 patients undergoing Brief Psychodynamic Investigation (4 sessions). Alliance development occurred rapidly and was clearly established by the third session. Neither defensive functioning nor supportive or exploratory interventions alone differentiated early alliance development. However, the degree of adjustment of therapists' interventions to patients' level of defensive functioning discriminated a low alliance from both improving and high alliances. The adjustment of therapeutic interventions to patients' level of defensive functioning is a promising predictor of alliance development and should be examined further, alongside other predictors of outcome. PMID- 11402079 TI - Supportive techniques: are they found in different therapies? AB - Therapists of different persuasions use various techniques. Although many of these techniques are specific to their theory of treatment, others are practiced in common among different forms of psychotherapy. Many of these common techniques have been previously described, but supportive techniques have been largely ignored. The authors distinguish between the use of supportive techniques and the therapeutic alliance. Using Luborsky's definition of supportive techniques, they examine the empirical literature on the use of these supportive techniques in various therapies. They conclude that supportive techniques are often used in different forms of psychotherapy or counseling. PMID- 11402080 TI - The factor structure of the working alliance inventory in cognitive-behavioral therapy. AB - Studies of the therapeutic alliance in cognitive-behavioral therapy (CBT) have varied in their results, necessitating a deeper understanding of this construct. Through an exploratory factor analysis of the alliance in CBT, as measured by the Working Alliance Inventory (shortened, observer-rated version), the authors found a two-factor structure of alliance that challenges the commonly accepted one general factor of alliance. The results suggest that the relationship between therapist and client (Relationship) may be largely independent of the client's agreement with and confidence in the therapist and CBT (Agreement/ Confidence), necessitating independent measures of these two factors, not one measure of a general alliance factor. PMID- 11402081 TI - Psychotherapy of adults with comorbid attention-deficit/hyperactivity disorder and psychoactive substance use disorder. AB - Psychotherapy for comorbid attention-deficit/ hyperactivity disorder (ADHD) and psychoactive substance use disorder (PSUD) is described. The authors suggest that relapse prevention is an appropriate initial treatment because it is well suited to manage both substance abuse and comorbid symptomatology such as impulsivity, distractibility, and avoidance associated with ADHD. Clinical vignettes describe typical interactions between patients and their therapists, highlighting opportunities for therapists to focus on overlapping symptoms. ADHD is one of the most common comorbid diagnoses with PSUD, and it is important that efficacious psychotherapies be developed to complement psychopharmacological approaches. Clinicians should consider psychotherapy as part of a multimodal treatment approach that includes medication and perhaps family therapy. Additional contributions from clinicians who have experience conducting psychotherapy with this population are needed in order to develop effective treatments. PMID- 11402082 TI - Putting suffering into perspective: implications of the patient's world view. AB - The need for suffering patients to reexamine their assumptions about life presents therapists with unique challenges and opportunities. Patients with a religious world view often struggle with whether God cares about, or has sent, their pain. Atheistic patients also search for the meaning in their lives but reject the answers offered by traditional authorities. Patients who are uncertain or ambivalent about their world view may challenge a therapist to provide an audience, insight, or direction. Using case examples, the author explores the therapist's role in helping patients with differing world views to integrate their suffering. PMID- 11402083 TI - Informed consent for case reports: the ethical dilemma of right to privacy versus pedagogical freedom. AB - A new international standard of editorial policy calls for written informed consent by the subject of every case report. Although this appears to be ethically appealing, the authors posit that in some situations, requesting informed consent may be unethical, can harm patients, and may erode the use of case reports as a valuable teaching method in psychiatry and psychotherapy. The authors discuss concerns regarding this new policy for mental health publication based on issues of transference, countertransference, best interest of the patient, and practicality. PMID- 11402085 TI - Development and characterization of novel erythropoiesis stimulating protein (NESP). AB - Studies on human erythropoietin (EPO) demonstrated that there is a direct relationship between the sialic acid-containing carbohydrate content of the molecule and its serum half-life and in vivo biological activity, but an inverse relationship with its receptor binding affinity. These observations led to the hypothesis that increasing the carbohydrate content, beyond that found naturally, would lead to a molecule with enhanced biological activity. Hyperglycosylated recombinant human EPO (rHuEPO) analogues were developed to test this hypothesis. Darbepoetin alfa (novel erythropoiesis stimulating protein, NESP), which was engineered to contain five N-linked carbohydrate chains (two more than rHuEPO), has been evaluated in preclinical animal studies. Due to its increased sialic acid-containing carbohydrate content, NESP is biochemically distinct from rHuEPO, having an increased molecular weight and greater negative charge. Compared with rHuEPO, it has an approximately 3-fold longer serum half-life, greater in vivo potency, and can be administered less frequently to obtain the same biological response. NESP is currently being evaluated in human clinical trials for treatment of anaemia and reduction in its incidence. PMID- 11402086 TI - An overview of the efficacy and safety of novel erythropoiesis stimulating protein (NESP). AB - Novel erythropoiesis stimulating protein (NESP, also known as darbepoetin alfa) is a molecule that stimulates erythropoiesis by the same mechanism as both native and recombinant human erythropoietin (rHuEPO). The extra sialic residues on NESP, however, allow it to be more stable in vivo with a 2- to 3-fold longer elimination half-life. Thus, following intravenous administration, the mean elimination half-life of NESP is 25.3 vs 8.5 h for rHuEPO. After subcutaneous administration, the mean terminal half-life for NESP is 48.8 h. The mean bioavailability of NESP after subcutaneous administration is approximately 37%, similar to that reported for rHuEPO. The pharmacokinetic data suggested that patients with renal anaemia would require less frequent dosing with NESP than with rHuEPO. NESP 0.45 microg/kg administered once weekly either intravenously or subcutaneously has been evaluated for the correction of chronic renal failure (CRF)-associated anaemia. The study population included CRF patients not receiving dialysis, along with those on haemodialysis or peritoneal dialysis. In patients who are rHuEPO-naive, NESP has a similar effect in correcting the anaemia as is seen with rHuEPO, but with less frequent dosing. Similarly, in patients previously receiving rHuEPO, NESP (whether administered intravenously or subcutaneously) is as effective as rHuEPO treatment for maintaining haemoglobin concentration when administered at a reduced frequency (i.e. either once weekly or once every other week). NESP is well tolerated, adverse effects are similar to those seen with rHuEPO, and no antibodies have been detected in >1500 patients exposed to NESP thus far. PMID- 11402087 TI - Practical guidelines for the use of NESP in treating renal anaemia. PMID- 11402089 TI - Morphogenesis of joint beta 2-microglobulin amyloid deposits. PMID- 11402090 TI - Advanced glycation and lipoxidation end products: reactive carbonyl compounds related uraemic toxicity. AB - Most studies on uraemic toxins have focused on disorders of enzymatic biochemistry. Recent studies to elucidate the pathogenesis of dialysis-related amyloidosis have provided new insights in progressive, irreversible protein modifications resulting from non-enzymatic biochemistry in uraemia. This paper focuses on two types of irreversible alterations of proteins: advanced glycation and lipoxidation. We investigate the causal role of various reactive carbonyl compounds (RCOs) accumulating in the serum, speculate on its clinical consequences, and discuss the therapeutic perspectives. PMID- 11402091 TI - Scintigraphic methods to detect beta2-microglobulin associated amyloidosis (Abeta2-microglobulin amyloidosis). AB - beta2-Microglobulin-derived amyloidosis (Abeta2m) represents a major cause or morbidity in patients with end-stage renal disease. Symptoms of Abeta2m amyloid are mainly related to (peri-) articular amyloid deposition. Conventional non invasive diagnostic techniques, i.e. clinical evaluation, joint ultrasonography or X-ray, computed tomography or magnetic resonance imaging findings, as well as conventional bone scans, suffer from relative non-specificity and/or low sensitivity. Two recent methods, namely scintigraphy with radiolabelled serum amyloid P component (SAP) or with the radiolabelled Abeta2m-precursor protein, beta2-microglobulin (beta2m), yield more specific information. Using (123)I labelled SAP, Abeta2m deposits have been visualized in several long-term haemodialysis (HD) patients. However, this scan did not show tracer accumulation in other frequently involved sites, such as hips or shoulders, but did frequently label the spleen, which is usually spared from Abeta2m deposits. Scanning with (131)I-labelled beta2m, in contrast, yielded tracer accumulations corresponding to the typical distribution pattern of Abeta2m. Specificity of this method was shown by several methods, and the sensitivity was found to markedly exceed that of combined clinical and radiological investigations. Recently, both the radiation exposure and the optical resolution of this latter scan have been further refined by substituting (131)I with (111)In. In a final step we generated recombinant human beta2m (rhbeta2m). (111)In-rhbeta2m again failed to show significant tracer accumulation over joint regions in patients on short-term HD without evidence of Abeta2m amyloidosis. In contrast, local tracer accumulations similar to those observed with natural, (111)In-labelled beta2m could be demonstrated in long-term HD patients with evidence of Abeta2m amyloidosis. In conclusion, scintigraphy for Abeta2m with (111)In-labelled rhbeta2m provides a homogenous and safe recombinant protein source, and allows for the sensitive and specific non-invasive detection of Abeta2m-amyloid deposits in dialysis patients. PMID- 11402092 TI - Efficacy of ultra-pure dialysate in the therapy and prevention of haemodialysis associated amyloidosis. PMID- 11402093 TI - Efficacy of choice of dialysis membrane. PMID- 11402094 TI - Efficacy of haemodiafiltration. PMID- 11402095 TI - Reduction of circulating beta2-microglobulin level for the treatment of dialysis related amyloidosis. AB - Dialysis-related amyloidosis (DRA) is a common complication associated with long term haemodialysis therapy. The elimination of beta2-microglobulin (beta2m), the major constituent of the amyloid fibrils in DRA, from circulation has been expected to bring some clinical benefit. Recently, a direct haemoperfusion method using selective beta2m absorption column to eliminate circulating beta2m has been introduced into clinical practice. According to a recently performed, prospective, multicentre study, joint pain, stiffness and daily activities were significantly improved in patients with established DRA after the introduction of selective beta2m absorption therapy. Meanwhile, although osteoarticular lesions progressed in the control group, there was no significant progression in the selective beta2m absorption therapy group. The absorptive affinity of the column for beta2m is not quite specific and therefore some other unknown uraemic toxins might be removed also. However, the improvement of joint pain and the ability to undertake daily activities showed reversed correlations against plasma beta2m clearance. Symptoms associated with the increased amount of extracorporeal circulation and increased economical burden are areas of concern for this therapy. In conclusion, selective beta2m absorption therapy was suggested to have the potential to ameliorate established DRA symptoms and simultaneously prevent its local development. The mechanism by which DRA symptoms are ameliorated remains obscure. PMID- 11402096 TI - Predicting medical intractability of epilepsy in children: how certain can we be? PMID- 11402097 TI - Carpal tunnel syndrome: complex issues with a "simple" condition. PMID- 11402099 TI - Early development of intractable epilepsy in children: a prospective study. AB - BACKGROUND: Little is known about early prediction of intractable epilepsy (IE) in children. Such information could help guide the early use of new therapies in selected patients. METHODS: Children with newly diagnosed epilepsy (n = 613) were prospectively identified from child neurology practices in Connecticut (1993- 1997) and followed-up for the occurrence of IE (failure of > or = 2 drugs, > or = 1 seizure/month, over 18 months) [corrected]. Etiology and epilepsy syndromes were classified per International League Against Epilepsy guidelines. RESULTS: The median follow-up is 4.8 years, and 599 (97.7%) have been followed for more than 18 months. Sixty children (10.0%) have met the criteria for IE, including 34.6% with cryptogenic/symptomatic generalized, 2.7% with idiopathic, 10.7% with other localization-related, and 8.2% with unclassified epilepsy (p < 0.0001). After multivariable adjustment for epilepsy syndrome, initial seizure frequency (p < 0.0001), focal EEG slowing (p = 0.02), and acute symptomatic or neonatal status epilepticus (p = 0.001) were associated with an increased risk of IE, and age at onset between 5 and 9 years was associated with a lowered risk (p = 0.03). The absolute number of seizures and unprovoked or febrile status epilepticus were not associated substantially with IE. CONCLUSIONS: Approximately 10% of children meet criteria for IE early in the course of their epilepsy. Cryptogenic/symptomatic generalized syndromes carry the highest risk and idiopathic syndromes the lowest. Half of IE occurs in children with nonidiopathic localization-related syndromes. Initial seizure frequency is highly predictive of IE. By contrast, absolute number of seizures and unprovoked or febrile status epilepticus are not. PMID- 11402100 TI - Distribution of partial seizures during the sleep--wake cycle: differences by seizure onset site. AB - OBJECTIVE: To evaluate the effects of sleep on partial seizures arising from various brain regions. METHODS: The authors prospectively studied 133 patients with localization-related epilepsy undergoing video-EEG monitoring over a 2-year period. Seizure type, site of onset, sleep/wake state at onset, duration, and epilepsy syndrome diagnosis were recorded. Periorbital, chin EMG, and scalp/sphenoidal electrodes were used. A subset of 34 patients underwent all night polysomnography with scoring of sleep stages. RESULTS: The authors analyzed 613 seizures in 133 patients. Forty-three percent (264 of 613) of all partial seizures began during sleep. Sleep seizures began during stages 1 (23%) and 2 (68%) but were rare in slow-wave sleep; no seizures occurred during REM sleep. Temporal lobe complex partial seizures were more likely to secondarily generalize during sleep (31%) than during wakefulness (15%), but frontal lobe seizures were less likely to secondarily generalize during sleep (10% versus 26%; p < 0.005). CONCLUSIONS: Partial-onset seizures occur frequently during NREM sleep, especially stage 2 sleep. Frontal lobe seizures are most likely to occur during sleep. Patients with temporal lobe seizures have intermediate sleep seizure rates, and patients with seizures arising from the occipital or parietal lobes have rare sleep-onset seizures. Sleep, particularly stage 2 sleep, promotes secondary generalization of temporal and occipitoparietal, but not frontal, seizures. These findings suggest that the hypersynchrony of sleep facilitates both initiation and propagation of partial seizures, and that effects of sleep depend in part on the location of the epileptic focus. PMID- 11402101 TI - Multiperspective follow-up of untreated carpal tunnel syndrome: a multicenter study. AB - OBJECTIVE: To assess the course of untreated carpal tunnel syndrome (CTS). METHODS: The Italian CTS Study Group prospectively followed up (10 to 15 months) 196 untreated patients (274 hands) with idiopathic CTS with multiple measurements of CTS. Baseline factors were used to predict the evolution of untreated CTS in multiple regression analysis. RESULTS: Comparison of baseline and follow-up data showed a significant spontaneous improvement of patient-oriented and neurophysiologic measurements. A significant correlation between evolution and initial severity of CTS was observed. CTS measurements improved in patients with more severe initial impairment whereas they worsened in patients with milder initial impairment. The main positive prognostic factor was short duration of symptoms. Similarly, spontaneous improvement was more frequently associated with young age. Conversely, baseline bilateral symptoms and positive Phalen predicted a poor prognosis. CONCLUSIONS: Some patients with CTS improve spontaneously without surgical treatment. PMID- 11402102 TI - Gastroenteritis-associated Guillain-Barre syndrome on the Caribbean island Curacao. AB - BACKGROUND: The number of patients with Guillain-Barre syndrome (GBS) who have been observed in Curacao, the Netherlands Antilles, may be increasing. METHODS: Clinical and serologic data were obtained from records of patients admitted between 1987 and 1999 and fulfilling National Institute of Neurological and Communicative Disorders and Stroke criteria for GBS. When possible, serum and stool samples were collected. The results were compared with a large Dutch epidemiologic study. RESULTS: The authors identified 49 patients, an overall crude incidence rate (IR) in Curacao of 2.53/100,000 inhabitants (95% CI 1.87 to 3.35) (Dutch study 1.18, rate ratio (RR) of 2.14, p < 0.001). The IR in Curacao increased from 1.62 in 1987 to 1991 to 3.10 in 1992 to 1999, RR 5.22 (95% CI 2.48 to 10.2, p = 0.02). The IR showed a curvilinear shape within a year. In comparison with the Dutch group, patients from Curacao had a more severe course of the disease, with a mortality rate of 23% (3.4% in the Dutch group, p < 0.001), a higher percentage of preceding gastroenteritis (p < 0.001), and less sensory involvement (p < 0.001). In 8 of 10 serum samples, evidence was found for a recent infection with Campylobacter jejuni. CONCLUSIONS: The authors found a steady increase in incidence of GBS over the years in association with a more pronounced seasonal preponderance and a more severe course. The clinical characteristics suggest a role for C jejuni. PMID- 11402103 TI - The earliest pathologic alterations in dysferlinopathy. AB - BACKGROUND: Dysferlinopathies are associated with proximal or distal muscular dystrophy. Dysferlin immunolocalizes to the muscle fiber periphery but does not associate with the dystrophin--glycoprotein complex; its function in humans, and the mechanism by which it causes muscle fiber injury, are not known. The authors therefore searched for pathogenetic clues by examining early abnormalities in nonnecrotic muscle fibers in dysferlinopathy. Five dysferlin-deficient patients were investigated. Weakness was distal in two, proximal in one, and both proximal and distal in two. Patient 5 was only mildly affected. METHODS: Immunoblot analysis, membrane attack complex (MAC) immunolocalization, and quantitative electron microscopy. RESULTS: In Patients 1 through 4, but not in 5, part or the entire surface of isolated nonnecrotic muscle fibers immunostained for MAC. Quantitative electron microscopy of 175 nonnecrotic muscle fibers revealed one or more of the following: 1) small (0.11 to 1.8 microm) plasmalemmal defects in 64% of fibers; 2) thickened basal lamina over some defects; 3) replacement of the plasma membrane by one to multiple layers of small vesicles in 57% of fibers; 4) papillary projections, frequently disintegrating, in 24 to 36% of fibers in Patients 1 through 4 but absent in fibers of Patient 5; 5) small subsarcolemmal vacuoles, some undergoing exocytosis, in 57% of fibers; and 6) infrequent subsarcolemmal regions containing rough endoplasmic reticulum and abundant free ribosomes. CONCLUSIONS: Dysferlin is likely required for maintaining the structural integrity of the muscle fiber plasma membrane, and plasma membrane injury is an early event in the pathogenesis of dysferlinopathy. MAC activation can participate in but is not an initial or primary event causing muscle fiber injury. PMID- 11402104 TI - An atypical intronic deletion widens the spectrum of mutations in hereditary spastic paraplegia. AB - OBJECTIVE: To identify the genetic mutation responsible for autosomal dominant spastic paraplegia (HSP) in a large family with a "pure" form of the disorder. BACKGROUND: The disease locus in most families with HSP is genetically linked to the SPG4 locus on chromosome 2p21-p22. Some of these families have mutations in the splice-site or coding regions of the spastin gene (SPAST). METHODS: Linkage and mutational analyses were used to identify the location and the nature of the genetic defect causing the disorder in a large family. After the disease phenotype was linked to the SPG4 locus, all 17 coding regions and flanking intronic sequences of SPAST were analyzed by single-strand conformation polymorphism analysis (SSCP) and compared between affected and normal individuals. Direct sequencing and subcloning methods were used to investigate incongruous mobility shifts. RESULTS: The genomic sequence of SPAST showed a heterozygous four--base pair deletion (delTAAT) near the 3' splice-site of exon three in all 11 affected individuals but not in 21 normal family members or in 50 unrelated controls (100 chromosomes). CONCLUSIONS: This study identifies an atypical intronic microdeletion in SPAST that causes HSP and widens the spectrum of genetic abnormalities that cause the disorder. PMID- 11402105 TI - MeCP2 mutations in children with and without the phenotype of Rett syndrome. AB - BACKGROUND: Rett syndrome (RTT) is a neurodevelopmental disorder caused by mutations in the X-linked methyl CpG binding protein 2 (MeCP2) gene. METHODS: One hundred sixteen patients with classical and atypical RTT were studied for mutations of the MeCP2 gene by using DHPLC and direct sequencing. RESULTS: Causative mutations in the MeCP2 gene were identified in 63% of patients, representing a total of 30 different mutations. Mutations were identified in 72% of patients with classical RTT and one third of atypical cases studied (8 of 25). The authors found 17 novel mutations, including a complex gene rearrangement found in one individual involving two deletions and a duplication. The duplication was identical to a region within the 3' untranslated region (UTR), and represents the first report of involvement of the 3' UTR in RTT. The authors also report the identification of MeCP2 mutations in two males; a Klinefelter's male with classic RTT (T158M) and a hemizygous male infant with a Xq27-28 inversion and a novel 32 bp frameshift deletion [1154(del32)]. Studies examining the relationship between mutation type, X-inactivation status, and severity of clinical presentation found significant differences in clinical presentation between different types of mutations. Mutations in the amino-terminus were significantly correlated with a more severe clinical presentation compared with mutations closer to the carboxyl-terminus of MeCP2. Skewed X-inactivation patterns were found in two asymptomatic carriers of MeCP2 mutations and six girls diagnosed with either atypical or classical RTT. CONCLUSION: This patient series confirms the high frequency of MeCP2gene mutations causative of RTT in females and provides data concerning the molecular basis for clinical variability (mutation type and position and X-inactivation patterns). PMID- 11402106 TI - Randomized controlled trial of interferon- beta-1a in secondary progressive MS: Clinical results. AB - BACKGROUND: The beneficial effect of interferon beta on exacerbations in relapsing-remitting MS has been demonstrated repeatedly, but results concerning disability vary. OBJECTIVE: This multicenter, randomized, parallel-group, placebo controlled study tested two doses of interferon beta-1a in patients with secondary progressive MS, which may include relapses but is dominated by accumulating disability. METHODS: A total of 618 patients received subcutaneous placebo or interferon beta-1a, 22 or 44 microg three times weekly for 3 years. Patients were assessed every 3 months. RESULTS: The primary outcome, time to confirmed progression in disability, was not significantly affected by treatment (hazard ratio, 0.83; 95% CI, 0.65 to 1.07; p = 0.146 for 44 microg versus placebo). Relapse rate was reduced from 0.71 per year with placebo to 0.50 per year with treatment (p < 0.001 for both doses). Significant treatment effects were seen on other exacerbation-related outcomes and on a composite measure incorporating five separate clinical and MRI outcomes. The hazard ratio for time to progression for the combined interferon beta-1a groups compared with placebo was 0.74 among patients reporting relapses in the 2 years before study (p = 0.055), and 1.01 for those without prestudy relapses (p = 0.934). An unexpected treatment-by-sex interaction favored women. The drug was well tolerated. CONCLUSIONS: Treatment with interferon beta-1a did not significantly affect disability progression in this cohort, although significant treatment benefit was observed on exacerbation-related outcomes. Exploratory post hoc analyses suggested greater benefit in women and in patients who had reported at least one relapse in the 2 years before the study. PMID- 11402107 TI - Randomized controlled trial of interferon-beta-1a in secondary progressive MS: MRI results. AB - OBJECTIVE: To examine MRI changes resulting from treatment of secondary progressive MS (SPMS) with two doses of interferon-beta-1a (Rebif). BACKGROUND: Interferon-beta (IFN-beta) reduces relapses and delays progression in relapsing remitting MS, but there are conflicting results on its clinical benefit in SPMS. METHODS: In a double-blind, randomized, multicenter, placebo-controlled study (SPECTRIMS), 618 patients received IFN-beta-1a 22 microg, 44 microg, or placebo subcutaneously three times weekly for 3 years. T2 activity and burden of disease (BOD) were measured in 617 patients by using semiannual proton density/T2 weighted (PD/T2) MRI scans. A cohort of 283 patients also had 11 monthly PD/T2 and T1-weighted gadolinium-enhanced (T1-Gd) scans at study start. RESULTS: Treatment reduced median numbers of active lesions per patient per scan (semiannual T2 activity: 0.17, 0.20 and 0.67 for the high dose, low dose, and placebo, p < 0.0001; monthly combined unique activity [T1+T2]: 0.11, 0.22, and 1.00, p < 0.0001) and accumulation of BOD (percent change from baseline to month 36: -1.3, -0.5, and 10.0 for the high dose, low dose, and placebo, respectively; p = 0.0001). MRI benefit was most evident in the subgroup of patients who reported relapses in the 2 years before the study. Neutralizing antibody development was associated with reduction in treatment effect: antibody-positive patients did not show significant differences from placebo at either dose. CONCLUSIONS: Interferon-beta-1a used in SPMS showed significant effects on all MRI measures, particularly in patients with relapses in the 2 years before the study. PMID- 11402108 TI - A randomized trial of intravenous immunoglobulin in inflammatory demyelinating optic neuritis. AB - OBJECTIVE: To determine whether IV immunoglobulin (IVIg) reverses chronic visual impairment in MS patients with optic neuritis (ON). METHODS: In this double blind, placebo-controlled Phase II trial, 55 patients with persistent acuity loss after ON were randomized to receive either IVIg 0.4 g/kg daily for 5 days followed by three single infusions monthly for 3 months, or placebo. RESULTS: The trial was terminated by the National Eye Institute because of negative results when 55 of the planned 60 patients had been enrolled. Fifty-two patients completed the scheduled infusions, and 53 patients completed 12 months of follow up. Analysis of this data indicated that a difference between treatment groups was not observed for the primary outcome measure, improvement in logMAR visual scores at 6 months (p = 0.766). Exploratory secondary analyses suggested that IVIg treatment was associated with improvement in visual function (including logMAR visual scores at 6 months and visual fields at 6 and 12 months) in patients with clinically stable MS during the trial. CONCLUSIONS: IVIg administration does not reverse persistent visual loss from ON to a degree that merits general use. PMID- 11402109 TI - A randomized, double masked, controlled trial of botulinum toxin type A in essential hand tremor. AB - OBJECTIVE: To evaluate the safety and efficacy of botulinum toxin type A injection in essential tremor of the hand. BACKGROUND: Botulinum toxin type A is an effective treatment for dystonia, spasticity, and other movement disorders and has been found to be useful in open-label studies and one double-masked study of essential hand tremor. METHODS: One hundred thirty-three patients with essential tremor were randomized to low-dose (50 U) or high-dose (100 U) botulinum toxin type A (Botox) or vehicle placebo treatment. Injections were made into the wrist flexors and extensors. Patients were followed for 16 weeks. The effect of treatment was assessed by clinical rating scales, measures of motor tasks and functional disability, and global assessment of treatment. Hand strength was evaluated by clinical rating and by a dynamometer. RESULTS: Both doses of botulinum toxin type A significantly reduced postural tremor on the clinical rating scales after 4 to 16 weeks. However, kinetic tremor was significantly reduced only at the 6-week examination. Measures of motor tasks and functional disability were not consistently improved with botulinum toxin type A treatment. Grip strength was reduced for the low- and high-dose botulinum toxin type A groups as compared with the placebo group. Adverse reactions consisted mainly of dose-dependent hand weakness. CONCLUSION: Botulinum toxin type A injections for essential tremor of the hands resulted in significant improvement of postural, but not kinetic, hand tremors and resulted in limited functional efficacy. Hand weakness is a dose-dependent significant side effect of treatment at the doses used in this study. PMID- 11402110 TI - CSF detection of the 14-3-3 protein in unselected patients with dementia. AB - OBJECTIVE: To determine the usefulness of the 14-3-3 test in patients with dementia of various causes. BACKGROUND: Recent reports have suggested that the detection of the 14-3-3 protein in the CSF of patients with Creutzfeldt--Jakob disease is a highly sensitive and specific marker of the disease that might be used as a diagnostic criterion. We examined the validity of this test when applied to a cohort of unselected patients prospectively examined for an ongoing dementing process. METHODS: One hundred patients underwent an extensive neurologic examination for dementia, including a CSF 14-3-3 protein immunoblotting assay. Final clinical diagnoses were compared with the qualitative results of the test, and statistical measures of test validity were carried out. RESULTS: We found a positive test in 14 of 100 patients, only two of whom had definite Creutzfeldt--Jakob disease. Positive results were found in patients with various degenerative dementias, including AD (4), frontotemporal dementia (2), and dementia with Lewy body (1), and in patients with vascular dementia (1), carcinomatous meningitis (1), and anoxic encephalopathy (1). In two other positive patients, the dementia could not be confidently classified. Sensitivity, specificity, and negative predictive value were fairly good, but positive predictive value was poor. Similar results were found independently of the disease duration. There was no correlation between intensity nor pattern of the 14-3-3 protein expression and diagnosis. CONCLUSIONS: The 14-3-3 test is not valid for discriminating between Creutzfeldt--Jakob disease and non-Creutzfeldt- Jakob disease in unselected patients with dementia. Positive results are found in various degenerative and secondary, prion-unrelated dementias. PMID- 11402111 TI - Memory decline in healthy older people: implications for identifying mild cognitive impairment. AB - BACKGROUND: Criteria for mild cognitive impairment require objective evidence of a memory deficit but do not require objective evidence of memory decline. Application of these criteria may therefore result in the misclassification of older patients with memory decline as normal because their neuropsychological test performance at a single point in time may be within normal limits. This study aimed to identify and characterize older people with memory decline. METHOD: Word list delayed recall (WLDR) test performance was assessed on five occasions during a 2-year period in a cohort of healthy older individuals. Older people with declining (n = 35) and nondeclining (n = 66) WLDR scores were identified. Both subgroups were then compared on apoE genotype, Clinical Dementia Rating, and neuropsychological test performance at the fifth assessment. RESULTS: Thirty-four percent of the group with declining memory recorded a Clinical Dementia Rating of 0.5, compared with 5% of the nondeclining memory group. No between-group differences were observed in cognitive domains other than memory, self-reported cognitive failures, or the proportion of each group carrying the apoE epsilon 4 allele. CONCLUSIONS: A large proportion of healthy older individuals show memory decline, which may represent the early stages of a potentially more severe cognitive impairment. Further investigation is necessary to determine the relationship between apoE genotype, self-reported cognitive impairment, and memory decline in older people. PMID- 11402112 TI - Cerebral white matter lesions and subjective cognitive dysfunction: the Rotterdam Scan Study. AB - OBJECTIVE: To determine the relationship between cerebral white matter lesions (WML) and subjective cognitive dysfunction. BACKGROUND: Subjective cognitive dysfunction is present when a person perceives failures of cognitive function. When annoying enough, these failures will be expressed as complaints. Subjective cognitive dysfunction may be a prelude to or coincide with objective cognitive impairment. WML have been related to objective cognitive impairment and dementia, but their relationship with subjective cognitive dysfunction is not clear. Previous population-based studies on the latter relationship have been limited in sample size, recording of subjective cognitive function, and assessment of WML severity. METHODS: We randomly sampled 1,049 elderly nondemented participants from the general population. Data on subjective cognitive dysfunction and its progression were derived from a 15-item questionnaire. Objective cognitive performance was assessed using a series of neuropsychological tests. WML were scored on MRI for periventricular and subcortical regions separately. RESULTS: WML were associated with more subjective cognitive failures. WML were more severe for participants reporting progression of these failures compared with participants without these failures, especially within participants with better than average cognitive performance (p = 0.008, for periventricular WML). Participants with severe WML reported progression of cognitive failures more than twice as often than did those with little or no WML. The relationship between the severity of WML and subjective cognitive failures was present for periventricular and subcortical WML. CONCLUSIONS: WML are associated with subjective cognitive failures and in particular with reporting progression of these failures, even in the absence of objective cognitive impairment. PMID- 11402113 TI - Presenilin-1-associated abnormalities in regional cerebral perfusion. AB - OBJECTIVE: To investigate the influence of the presenilin-1 gene (PS-1) mutation on regional cerebral perfusion, SPECT was evaluated in 57 individuals. The subjects were members of a large pedigree from Colombia, South America, many of whom carry a PS-1 mutation for early-onset AD. METHODS: Members of this large kindred who were cognitively normal and did not carry the PS-1 mutation (n = 23) were compared with subjects who were carriers of the mutation but were asymptomatic (n = 18) and with individuals with the mutation and a clinical diagnosis of AD (n = 16). Cerebral perfusion was measured in each subject using hexamethylpropyleneamine oxime SPECT. The data were analyzed in two ways: 1) Mean cerebral perfusion in each of 4320 voxels in the brain was compared among the groups using t-tests (t-maps); and 2) each individual received a weighted score on 20 vectors (factors), based on a large normative sample (n = 200), using a method known as singular value decomposition (SVD). RESULTS: Based on t-maps, subjects with the PS-1 mutation who were asymptomatic demonstrated reduced perfusion in comparison with the normal control subjects in the hippocampal complex, anterior and posterior cingulate, posterior parietal lobe, and anterior frontal lobe. The AD patients demonstrated decreased perfusion in the posterior parietal and superior frontal cortex in comparison with the normal control subjects. Discriminant function analysis of the vector scores derived from SVD (adjusted for age and gender) accurately discriminated 86% of the subjects in the three groups (p < 0.0005). CONCLUSION: Regional cerebral perfusion abnormalities based on SPECT are detectable before development of the clinical symptoms of AD in carriers of the PS-1 mutation. PMID- 11402114 TI - CNS manifestations of Nasu-Hakola disease: a frontal dementia with bone cysts. AB - BACKGROUND: Nasu-Hakola disease or polycystic lipomembranous osteodysplasia with sclerosing leukoencephalopathy (PLOSL) is a genetically heterogeneous disease characterized by a combination of systemic bone cysts and dementia. OBJECTIVE: The authors present a neurologic, neuroradiologic, and neuropathologic analysis of a series of PLOSL patients in which the diagnosis has been confirmed by molecular genetic methods. METHODS: Clinical, neurophysiologic, and imaging follow-up data on eight patients as well as autopsy samples of three patients were analyzed in this study. All eight patients were homozygous for a loss-of function mutation in the DAP12 gene. RESULTS: In most patients, the disease debuted with pain in ankles and wrists after strain during the third decade, followed by fractures caused by cystic lesions in the bones of the extremities. Frontal lobe syndrome and dementia began to develop by age 30, leading to death by age 40. Neuroimaging disclosed abnormally high and progressively increasing bicaudate ratios and calcifications in the basal ganglia as well as increased signal intensities of the white matter on T2-weighted MR images even before the appearance of clinical neurologic symptoms. Three patients who had undergone autopsies showed an advanced sclerosing leukoencephalopathy with frontal accentuation, widespread activation of microglia, and microvascular changes. CONCLUSIONS: Although PLOSL in most patients manifests by bone fractures, some patients do not show any osseous symptoms and signs before the onset of neurologic manifestations. Consequently, patients with frontal-type dementia of unknown origin should be investigated by x-ray of ankles and wrists. The current results suggest early basal ganglia involvement in PLOSL. PMID- 11402115 TI - Influence of L-dopa and pramipexole on striatal dopamine transporter in early PD. AB - BACKGROUND: Animal data indicate that chronic exposure to dopaminergic drugs can alter levels of the dopamine transporter (DAT), which is critically involved in regulation of synaptic dopamine levels. DAT changes could influence the response to therapy in PD. METHODS: A randomized, assessor-blinded, placebo-controlled clinical trial was performed in subjects with early PD to determine whether L dopa or pramipexole might regulate striatal DAT binding as measured by PET with [(11)C]RTI-32. Thirty clinically asymmetrical patients were randomly assigned to receive 6 weeks of L-dopa (300/75 mg/d), pramipexole (1.5 mg/d), or placebo; PET studies were performed before and after treatment. RESULTS: Mean interval change in DAT binding was significantly reduced by 16% to 22% in all striatal regions (caudate, anterior and posterior putamen) of the L-dopa-treated patients, whereas significant changes in the pramipexole-treated patients were limited to the contralateral caudate (-15%), ipsilateral anterior putamen (-14%), and posterior putamen (-20%). In the placebo group there were significant changes in contralateral caudate (-11%) and ipsilateral anterior putamen (-12%). L-dopa and pramipexole produced similar clinical benefit. CONCLUSIONS: Short-term therapy with L-dopa and, to a lesser extent, pramipexole can modestly down-regulate striatal DAT in patients with early PD. Decreased striatal DAT could increase dopaminergic neurotransmission with potential benefit, but might also play a role in the development of dopamine-related response fluctuations in patients with advanced disease. Our data also suggest caution in interpretation of longitudinal imaging studies employing DAT to assess disease progression and the efficacy of neuroprotective agents. PMID- 11402116 TI - Local vs systemic corticosteroids in the treatment of carpal tunnel syndrome. AB - The authors compared the effectiveness of low-dose, short-term oral prednisolone vs local methylprednisolone injection in a prospective, double-blinded, parallel treatment study of carpal tunnel syndrome (CTS). A single injection of 15 mg methylprednisolone resulted in significant improvement in global symptom scores over a 12-week period. This study demonstrated the superiority of local steroid injection to oral steroid in the treatment of CTS. PMID- 11402117 TI - The frequency of carpal tunnel syndrome in computer users at a medical facility. AB - A survey was done of employees who were identified as frequent computer users. Although 29.6% of the employees reported hand paresthesias, only 27 employees (10.5%) met clinical criteria for carpal tunnel syndrome, and in 9 (3.5%) the syndrome was confirmed by nerve conduction studies. Affected and unaffected employees had similar occupations, years using a computer, and time using the computer during the day. The frequency of carpal tunnel syndrome in computer users is similar to that in the general population. PMID- 11402118 TI - Large-fiber neuropathy in distal sensory neuropathy with normal routine nerve conduction. AB - Near-nerve needle sensory nerve conduction of plantar nerves in 100 patients with distal sensory neuropathy with normal routine nerve conduction (DSN-NNC) found the definite neuropathy pattern (abnormality in more than three of six tested nerves) in 65%, axonal neuropathy in 35%, and the known cause in 37% of patients. Absent or diminished reflexes were a reliable indicator for large fiber neuropathy (LFN). This near-nerve needle plantar nerve study provides useful and unequivocal evidence of its value in identifying neuropathy in DSN-NNC by finding LFN in 65% of patients. PMID- 11402119 TI - Parkin gene causing benign autosomal recessive juvenile parkinsonism. AB - Autosomal recessive juvenile parkinsonism (AR-JP) is an early-onset parkinsonism caused by exonic deletions or point mutations in the parkingene. The relationship between the type of the genetic defect and the clinical presentation, the response to therapy, and the evolution have not been yet determined. The authors describe a single-basepair deletion at nucleotide 202 in exon 2 of the parkin gene in a kindred with a benign clinical course. PMID- 11402120 TI - Lesion of thalamic centromedian--parafascicular complex after chronic deep brain stimulation. AB - A patient with PD who exhibited disabling tremor and prominent dyskinesia underwent deep brain stimulation (DBS) of the left thalamic ventral intermediate nucleus. The electrode migrated and was replaced but with suboptimal clinical response. Two years later, postmortem analysis found the second electrode tip had entered the thalamic centromedian-parafascicular complex. There was a small thalamotomy and cell loss exceeding that found in PD. Thalamic damage may occur in association with DBS for PD. PMID- 11402121 TI - Prevalence of primary blepharospasm in a community of Puglia region, Southern Italy. AB - The authors ascertained the prevalence of primary blepharospasm (BSP) in a community located in Puglia, a region in Southern Italy, by focusing on neurologists and ophthalmologists. The crude prevalence rates were 133 per million (95% CI, 61--153) for both focal and segmental BSP, 74 per million (95% CI, 24--173) for focal BSP, and 59 per million (95% CI, 16--151) for segmental BSP. Prevalence rate increased with age. Apraxia of eyelid opening and BSP coexisted in one third of cases. PMID- 11402122 TI - Thrombolysis of extracranial and intracranial arteries after IV abciximab. AB - Abciximab was administered intravenously to three patients to treat thrombus associated with atherosclerotic stenosis of a supra-aortic artery. In two cases, the thrombi had been identified on a previous angiography more than a week before treatment. Successful thrombolysis was achieved with abciximab in all three cases. In this small series, abciximab had a thrombolytic effect on established thrombi that were refractory to previous antithrombotic treatment attempts. PMID- 11402123 TI - Rescue treatment with abciximab in acute ischemic stroke. AB - In acute ischemic stroke, reocclusion after an initially successful thrombolysis treatment can occur and is associated with increased morbidity and mortality. The authors present the successful use of abciximab, a platelet glycoprotein IIb/IIIa receptor inhibitor, in a patient with a thrombotic occlusion of the proximal middle cerebral artery, which was refractory to combined IV and intra-arterial thrombolysis and percutaneous intracranial balloon angioplasty. PMID- 11402124 TI - Unilateral swollen disc due to increased intracranial pressure. AB - Truly unilateral papilledema is rare and poses a diagnostic problem. The authors have prospectively looked for patients with truly unilateral papilledema and found 15 patients, 10 of whom had idiopathic intracranial hypertension. Neuroimaging did not indicate a reason for the lack of swelling in the other nerve. The visual deficits and outcomes were similar to those of patients with bilateral papilledema. Although monocular papilledema is uncommon, a lumbar puncture with opening pressure measurement should be considered. PMID- 11402125 TI - Postictal language dysfunction in complex partial seizures: effect of contralateral ictal spread. AB - The authors report postictal language evaluation in patients monitored with bitemporal depth electrodes. Patients whose seizures began in the nondominant temporal lobe and propagated to the contralateral temporal lobe had a prolonged postictal language delay (PILD) with paraphasic errors compared with seizures that did not spread. Shorter propagation time was also associated with a longer PILD. Our study suggests that ictal involvement of the dominant temporal lobe is important in postictal language behavior. PMID- 11402126 TI - The angiotensin I converting enzyme gene as a susceptibility factor for dementia. AB - The frequency of a genetic susceptibility vascular factor, the deletion (D) allele of the angiotensin I converting enzyme gene (ACE), coding for a key enzyme of the renin angiotensin system, was characterized in two independent case- control studies. The results of the current study suggest that bearing at least one ACE D allele is a risk factor to develop dementia for subjects older than 74 years. This observation reinforces the hypothesis of a major implication of vascular risk factors in the occurrence of all types of dementia. PMID- 11402127 TI - Rate of cognitive decline in AD is accelerated by the interleukin-1 alpha -889 *1 allele. AB - The reason for differences in rate of cognitive decline in AD is unknown. The interleukin-1 alpha (IL-1 alpha) -889 *2 allele is associated with increased risk for AD. Surprisingly, in a sample of 114 patients followed for an average of 3.8 years, individuals homozygous for the IL-1 alpha -889 *1 allele declined significantly more rapidly on the Mini-Mental State Examination than did others. There was no difference in rate of decline between patients with and without the APOE epsilon 4 allele. These results support the hypothesis that inflammation is important in the clinical course of AD. PMID- 11402128 TI - Hypothalamic amnesia with spontaneous confabulations: a clinicopathologic study. AB - Previous studies demonstrated that patients producing spontaneous confabulations fail to suppress currently irrelevant memory traces and have anterior limbic lesions, particularly involving the orbitofrontal cortex or the basal forebrain. Here, a woman is described who had sarcoidosis damaging the medial hypothalamus and endocrine dysfunction, and a severe memory failure characterized by spontaneous confabulation, disorientation, and severely impaired free recall with preserved recognition. Isolated hypothalamic damage may produce the same type of memory disorder as orbitofrontal damage. PMID- 11402129 TI - Epileptic intermittent snoring. PMID- 11402130 TI - Dopaminergic changes in human brain following acute exposure to gamma hydroxybutyrate. PMID- 11402131 TI - Acute motor radiculopathy as a first symptom of MS: anatomic-radiologic correlation. PMID- 11402132 TI - The Babinski-Nageotte syndrome. PMID- 11402133 TI - Clinical neurophysiology training and certification in the United States: 2000. PMID- 11402134 TI - Pyridostigmine-induced microcephaly. PMID- 11402135 TI - Syndrome of cerebral spinal fluid hypovolemia: clinical and imaging features and outcome. PMID- 11402138 TI - Multifocal motor neuropathy improved by IVIg. PMID- 11402136 TI - Cauda equina syndrome from intradiscal electrothermal therapy. PMID- 11402141 TI - A multidisciplinary approach to Pick's disease and frontotemporal dementia. Conference proceedings. May 6-7, 1999. Philadelphia, Pennsylvania, USA. PMID- 11402142 TI - Pick's disease: a clinical overview. AB - What is to be understood by the term Pick's disease? Is this a clinical syndrome(s) of frontotemporal lobar atrophy, or a more specific clinicopathological concept of frontotemporal lobar atrophy with Pick bodies and/or Pick cells on neuropathology? The author discusses these concepts in an historical context as an introduction to this symposium. PMID- 11402143 TI - Frontotemporal dementia (Pick's disease): clinical features and assessment. AB - The clinical presentation in frontotemporal dementia (FTD) reflects the distribution of the pathologic changes rather than the exact histologic subtype of the disease. Three major clinical syndromes can be identified: 1) frontal variant FTD (dementia of frontal type) in which changes in social behavior and personality predominate, reflecting the orbitobasal frontal lobe focus of the pathology. Traditional cognitive tests are insensitive, but more specific measures are under development; 2) semantic dementia (progressive fluent aphasia) in which there is a breakdown in the conceptual database which underlies language production and comprehension, although deficits in nonverbal semantic knowledge can also be shown on neuropsychologic testing. Patients with semantic dementia have asymmetric anterolateral temporal atrophy with relative sparing of the hippocampal formation, which is typically worse on the left side. A variant of this syndrome affecting the right temporal lobe presents with progressive prosopagnosia; 3) progressive nonfluent aphasia in which the phonologic and syntactic components of language are affected in association with left peri Sylvian atrophy. The assessment of patients with potential FTD involves a multidisciplinary approach. The development of comprehensive caregiver-based neuropsychiatric instruments, neuropsychologic tasks sensitive to semantic memory and other key cognitive impairments, and functional (hexamethyly-propyleneamine SPECT) and structural (MRI) brain imaging represent significant advances in the field. PMID- 11402144 TI - The influence of right frontotemporal dysfunction on social behavior in frontotemporal dementia. AB - BACKGROUND: Frontotemporal dementia (FTD) is associated with a variety of cognitive and behavioral dysfunctions. Symptoms may be influenced by the relative involvement of the right versus the left hemisphere, with left-sided FTD manifesting language changes and right-sided FTD presenting with aggressive, antisocial, and other socially undesirable behaviors. OBJECTIVE: To test the hypothesis that right-sided FTD is associated with socially undesirable behavior. METHODS: The authors assessed 41 patients with FTD diagnosed by the new research criteria for FTD(1) including behavioral, neuropsychologic, and neurologic testing as well as SPECT and MRI. Based on visual inspection of SPECT scans, 12 patients were classified as having predominantly right-sided and 19 patients were classified as having predominantly left-sided FTD. A clinician blinded to the imaging data reviewed medical records to tabulate the frequency of the following socially undesirable behaviors: criminal behavior, aggression, loss of job, alienation from family/friends, financial recklessness, sexually deviant behavior, and abnormal response to spousal crisis. RESULTS: Eleven of 12 right sided and 2 of 19 left-sided FTD patients had socially undesirable behavior as an early presenting symptom (chi = 23.3, p < 0.001). CONCLUSION: The authors conclude that right-sided frontotemporal degeneration is associated with socially undesirable behavior. The early presence of socially undesirable behavior in FTD differentiates right-sided from left-sided degeneration. The results highlight the importance of the right hemisphere, especially frontotemporal regions, in the mediation of social behavior. The potential mechanism for these social losses with right-sided disease is discussed. PMID- 11402145 TI - Neuropathology of Pick's disease. AB - The author reviews the gross, microscopic, and biochemical pathology associated with classic Pick's disease, as defined by Constantinidis' Type A Pick's disease. This is contrasted with other Pick's disease subtypes, including Constantinidis' Type B (corticobasal degeneration) and Type C (dementia lacking distinctive histology). PMID- 11402146 TI - Missense and splice site mutations in tau associated with FTDP-17: multiple pathogenic mechanisms. AB - Recent identification of mutations in the gene encoding the microtubule associated protein tau in the inherited frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17) has demonstrated that tau dysfunction can lead to neurodegeneration. At least nine missense mutations and one deletion mutation (DeltaK280) have been identified in exons 9 through 13 that encode the microtubule-binding domains of tau. In addition, five mutations have been found close to the 5' splice site of exon 10. The FTDP-17 missense and splice site mutations have multiple effects on the biology and function of tau. It is likely that these varied pathogenic mechanisms explain the wide range of clinical and neuropathologic features observed in the FTDP-17 tauopathies. PMID- 11402147 TI - Transgenic mouse models of tauopathies: prospects for animal models of Pick's disease. AB - Because filamentous neuronal tau inclusions are neuropathologic hallmarks of Pick's disease and a number of other neurodegenerative disorders known as tauopathies, the authors generated lines of transgenic (Tg) mice that overexpress the shortest human tau isoform in the CNS. These Tg mice accumulated argyrophilic and filamentous tau immunoreactive neuronal inclusions with age in cortex and brainstem as well as in spinal cord, where these inclusions were most abundant and associated with gliosis. The ventral roots of affected spinal cord segments showed axonal degeneration, whereas residual axons exhibited reduced microtubules and reduced fast axonal transport. The inclusions were composed of 10 to 20 nm tau immunopositive straight filaments. In addition, the Tg mice developed age related motor weakness as well as progressive hyperphosphorylation and decreased solubility of brain and spinal cord tau proteins. Thus, these Tg mice recapitulate phenotypic features of human tauopathies. In this article the authors review the phenotype of these Tg mice and discuss how the availability of relevant animal model of tauopathies will facilitate discovery of more effective therapies to treat Pick's disease and related disorders characterized by filamentous tau pathology in selectively vulnerable regions of the CNS. PMID- 11402148 TI - Frontotemporal dementia: report of a familial case. AB - The authors describe a 49-year-old woman (R.K.) who presented with one year of progressive frontal lobe dysfunction, including signs of expressive aphasia. Signs of parkinsonism were absent until late in the clinical course. Neuropsychologic testing and neuroimaging studies are described. The patient died at age 55, after 7 years of symptoms. Family history was remarkable for a mother who died at the age of 45, after experiencing 7 years of progressive aphasia. R.K.'s brain showed asymmetric frontotemporal atrophy, which was more severe on the left side. Histopathologic analysis was remarkable for numerous tau-positive neurons with some classic-appearing Pick bodies and many ballooned neurons. Tau positive glial cells were also present. The authors suggest that the abnormal tau aggregates are related to the symptoms experienced by affected members of this family. PMID- 11402149 TI - Bcl-2 family protein behavior in frontotemporal dementia implies vascular involvement. AB - Frontotemporal dementia (FTD) is a neurodegenerative disease associated with aging for which the etiology is unclear. Relatively little is known about the pathology of this disease, which has only recently been a topic of investigation for dementia researchers. Though the known pathology of FTD includes neuron loss, the mechanism of neuronal death is not known. In this study, the authors investigated apoptotic pathways as a possible mechanism of neuronal cell death in FTD. They evaluated immunoreactivity for Bcl-2 family protein members Bcl-x and Bax in postmortem frontal cortex from FTD, AD, and control cases. Bcl-x(L), Bcl x(S), and Bax all exhibited altered immunoreactivity in FTD cases as compared with control cases. Bcl-x immunoreactivity varied widely among both controls and FTD cases. However, Bcl-x(L) showed strong white matter immunoreactivity in all FTD cases, whereas white matter immunoreactivity was absent in controls. These trends in Bcl-x immunoreactivity suggest a strong white matter involvement in the pathology of FTD. Bax immunoreactivity also varied across all cases. Bax immunoreactivity was observed in terminal transferase dUTP nick ending labeling (TUNEL) positive neurons in both FTD and AD cases. However, one notable finding was immunoreactivity to Bax in astrocytes of FTD cases, as well as endothelial cells of the cerebrovasculature. Neither astrocytic nor endothelial cell immunoreactivity to Bax was exhibited in control or AD cases. Because Bax is a pro-apoptotic protein, this finding suggests the presence of a cerebrovascular component in the pathology of FTD. These findings, coupled with the proposed functions of the Bcl-2 family proteins, suggest that an apoptotic pathway may be responsible for neuron, and possibly astrocyte, death in FTD. PMID- 11402150 TI - Therapy and management of frontal lobe dementia patients. AB - Patients with frontal lobe dementia (FLD) include those who suffer from Pick's disease, corticobasal degeneration, FLD without specific histopathologic features, as well as the infrequent families with frontotemporal dementia and parkinsonism associated with chromosome 17. Currently there have been no systematic efforts to manage and to treat patients with FLD. Drawing on the accumulated experience of clinicians and the known therapeutic approaches for patients with other neurodegenerative disorders such as AD and PD, the author discusses possible neurotransmitter replacement and biologic therapeutic approaches for patients with FLD. PMID- 11402151 TI - Behavior and treatment in frontotemporal dementia. AB - Behavioral syndromes are central to the clinical presentation of frontotemporal dementia. The authors review the behavioral changes seen in frontotemporal dementia and describe pharmacologic interventions for these behavioral syndromes. PMID- 11402152 TI - Responding to safety issues in frontotemporal dementias. AB - As frontotemporal dementia progresses in individuals, safety issues related to behaviors and injury become a paramount concern. In addition to self-care deficits, frontotemporal dementias are often characterized by behavioral manifestations that include aggression and disinhibition. These behaviors may place the patient and caregivers at risk of injury, stress, and social embarrassment, and frequently lead to institutionalization. Additionally, motor disturbances associated with frontotemporal dementias may contribute to risk of injury from falls. The authors present an integrated biopsychosocial model to guide assessment of needs that may be expressed through behavior. Environmental, behavioral, and psychosocial strategies to assist caregivers in preventing and responding to behaviors and risks are discussed, with the goal of promoting maximum function and quality of life and minimizing caregiver strain. The authors discuss the dangers of physical restraints, which are commonly suggested as a response to fall risk and behavioral symptoms without an awareness of research based data regarding their lack of efficacy. Benefits and risks of a variety of need-based interventions are presented in a practical, clinically relevant manner. The discussion of diverse safety-enhancing interventions is intended to enable clinicians and caregivers to identify individualized care strategies for patients with frontotemporal dementia. PMID- 11402153 TI - Rehabilitation applications in caring for patients with Pick's disease and frontotemporal dementias. AB - The basic principles of rehabilitation philosophy relevant to the care of patients with neurodegenerative diseases, specifically Pick's disease and frontotemporal dementias, are briefly defined. These principles are illustrated by the case study of a patient with a clinical diagnosis of Pick's disease. PMID- 11402154 TI - An algorithm (decision tree) for the management of Parkinson's disease (2001): treatment guidelines. PMID- 11402155 TI - A role for PsbZ in the core complex of photosystem II. PMID- 11402156 TI - Functional evolutionary genetics and plant adaptation: linking phenotype and genotype. PMID- 11402157 TI - An evolutionist visits the 43rd annual maize genetics conference. PMID- 11402158 TI - Uncovering secretory secrets: inhibition of endoplasmic reticulum (ER) glucosidases suggests a critical role for ER quality control in plant growth and development. PMID- 11402159 TI - The arabidopsis serrate gene encodes a zinc-finger protein required for normal shoot development. AB - Organogenesis in plants depends upon the proper regulation of many genes, but how such necessary changes in gene expression are coordinated is largely unknown. The serrate (se) mutant of Arabidopsis displays defects in the initiation and elaboration of cotyledons and post-embryonic lateral organs. Cloning the SE gene revealed that it encodes a protein with a single, C(2)H(2)-type, zinc finger related to genes in other eukaryotes. Consistent with a role in organogenesis, the SE gene is transcribed in shoot meristems and in emerging organ primordia throughout development. Expression of the SE cDNA under the control of a heterologous promoter caused both accelerated and arrested plant growth, and these phenotypes were due to overexpression and co-suppression of the SE gene, respectively. Our analysis of the se mutant and the SE gene suggests a role for the SE gene product in regulating changes in gene expression via chromatin modification. Consistent with this proposed function, a synergistic double mutant phenotype was seen for plants mutant at both the SE locus and the locus encoding the largest subunit of chromatin assembly factor I. PMID- 11402160 TI - EARLY FLOWERING3 encodes a novel protein that regulates circadian clock function and flowering in Arabidopsis. AB - Higher plants use photoperiodic cues to regulate many aspects of development, including the transition from vegetative to floral development. The EARLY FLOWERING3 (ELF3) gene is required for photoperiodic flowering and normal circadian regulation in Arabidopsis. We have cloned ELF3 by positional methods and found that it encodes a novel 695-amino acid protein that may function as a transcriptional regulator. ELF3 transcript level is regulated in a circadian manner, as is expected of a zeitnehmer input pathway component. Overexpression of the LATE ELONGATED HYPOCOTYL gene, which has been proposed to function as a clock component, did not abolish circadian regulation of ELF3 transcription, providing further evidence that ELF3 is a circadian clock input pathway component. PMID- 11402161 TI - ELF3 encodes a circadian clock-regulated nuclear protein that functions in an Arabidopsis PHYB signal transduction pathway. AB - Many aspects of plant development are regulated by photoreceptor function and the circadian clock. Loss-of-function mutations in the Arabidopsis EARLY FLOWERING 3 (ELF3) and PHYTOCHROME B (PHYB) genes cause early flowering and influence the activity of circadian clock-regulated processes. We demonstrate here that the relative abundance of the ELF3 protein, which is a novel nucleus-localized protein, displays circadian regulation that follows the pattern of circadian accumulation of ELF3 transcript. Furthermore, the ELF3 protein interacts with PHYB in the yeast two-hybrid assay and in vitro. Genetic analyses show that ELF3 requires PHYB function in early morphogenesis but not for the regulation of flowering time. This suggests that ELF3 is a component of a PHYB signaling complex that controls early events in plant development but that ELF3 and PHYB control flowering via independent signal transduction pathways. PMID- 11402162 TI - ELF3 modulates resetting of the circadian clock in Arabidopsis. AB - The Arabidopsis early flowering 3 (elf3) mutation causes arrhythmic circadian output in continuous light, but there is some evidence of clock function in darkness. Here, we show conclusively that normal circadian function occurs with no alteration of period length in elf3 mutants in dark conditions and that the light-dependent arrhythmia observed in elf3 mutants is pleiotropic on multiple outputs normally expressed at different times of day. Plants overexpressing ELF3 have an increased period length in both constant blue and red light; furthermore, etiolated ELF3-overexpressing seedlings exhibit a decreased acute CAB2 response after a red light pulse, whereas the null mutant is hypersensitive to acute induction. This finding suggests that ELF3 negatively regulates light input to both the clock and its outputs. To determine whether ELF3's action is phase dependent, we examined clock resetting by using light pulses and constructed phase response curves. Absence of ELF3 activity causes a significant alteration of the phase response curve during the subjective night, and constitutive overexpression of ELF3 results in decreased sensitivity to the resetting stimulus, suggesting that ELF3 antagonizes light input to the clock during the night. The phase of ELF3 function correlates with its peak expression levels in the subjective night. ELF3 action, therefore, represents a mechanism by which the oscillator modulates light resetting. PMID- 11402163 TI - Antisense expression of a cell wall-associated protein kinase, WAK4, inhibits cell elongation and alters morphology. AB - The Arabidopsis cell wall-associated receptor-like kinase (WAK) gene family contains five highly related members whose products are suited for exchanging signals between the intracellular and extracellular compartments. WAK members are expressed in specific organs and regulated differentially by various biotic and abiotic factors. To gain further insight into how WAKs function during development, we used a glucocorticoid-inducible system to express ectopically the WAK4 antisense gene. The induced expression of the WAK4 antisense gene resulted in a significant decrease of WAK proteins. Ninety-six hours after the induction of WAK4 antisense expression, WAK proteins became undetectable. Cell elongation was impaired, and lateral root development was blocked. The level of WAK protein could be controlled by the concentration of the applied inducer, dexamethasone, and was correlated with the severity of the cell elongation inhibition phenotype. These results suggest that the WAKs serve a vital role in cell elongation and are required for plant development. PMID- 11402164 TI - ADF proteins are involved in the control of flowering and regulate F-actin organization, cell expansion, and organ growth in Arabidopsis. AB - Based mostly on the results of in vitro experiments, ADF (actin-depolymerizing factor) proteins are thought to be key modulators of the dynamic organization of the actin cytoskeleton. The few studies concerned with the in vivo function of ADF proteins that have been reported to date were performed almost exclusively using single-cell systems and have failed to produce consistent results. To investigate ADF functions in vivo and during the development of multicellular organs, we generated transgenic Arabidopsis plants that express a cDNA encoding an ADF protein (AtADF1) in the sense or the antisense orientation under the control of a strong constitutively active promoter. Selected lines with significantly altered levels of AtADF protein expression were characterized phenotypically. Overexpression of AtADF1 resulted in the disappearance of thick actin cables in different cell types, caused irregular cellular and tissue morphogenesis, and reduced the growth of cells and organs. In contrast, reduced AtADF expression promoted the formation of actin cables, resulted in a delay in flowering, and stimulated cell expansion as well as organ growth. These results are consistent with the molecular functions of ADF as predicted by in vitro studies, support the global roles of ADF proteins during the development of a multicellular organism, and demonstrate that these proteins are key regulators of F-actin organization, flowering, and cell and organ expansion in Arabidopsis. PMID- 11402165 TI - The chloroplast gene ycf9 encodes a photosystem II (PSII) core subunit, PsbZ, that participates in PSII supramolecular architecture. AB - We have characterized the biochemical nature and the function of PsbZ, the protein product of a ubiquitous open reading frame, which is known as ycf9 in Chlamydomonas and ORF 62 in tobacco, that is present in chloroplast and cyanobacterial genomes. After raising specific antibodies to PsbZ from Chlamydomonas and tobacco, we demonstrated that it is a bona fide photosystem II (PSII) subunit. PsbZ copurifies with PSII cores in Chlamydomonas as well as in tobacco. Accordingly, PSII mutants from Chlamydomonas and tobacco are deficient in PsbZ. Using psbZ-targeted gene inactivation in tobacco and Chlamydomonas, we show that this protein controls the interaction of PSII cores with the light harvesting antenna; in particular, PSII-LHCII supercomplexes no longer could be isolated from PsbZ-deficient tobacco plants. The content of the minor chlorophyll binding protein CP26, and to a lesser extent that of CP29, also was altered substantially under most growth conditions in the tobacco mutant and in Chlamydomonas mutant cells grown under photoautotrophic conditions. These PsbZ dependent changes in the supramolecular organization of the PSII cores with their peripheral antennas cause two distinct phenotypes in tobacco and are accompanied by considerable modifications in (1) the pattern of protein phosphorylation within PSII units, (2) the deepoxidation of xanthophylls, and (3) the kinetics and amplitude of nonphotochemical quenching. The role of PsbZ in excitation energy dissipation within PSII is discussed in light of its proximity to CP43, in agreement with the most recent structural data on PSII. PMID- 11402166 TI - Nodule-specific regulation of phosphatidylinositol transfer protein expression in Lotus japonicus. AB - Phosphatidylinositol transfer proteins (PITPs) modulate signal transduction pathways and membrane-trafficking functions in eukaryotes. Here, we describe the characterization of a gene family from Lotus japonicus that encodes a novel class of plant PITP-like proteins (LjPLPs) and that is regulated in an unusual nodule specific manner. Members of this gene family were identified based on their nucleotide sequence homology with a previously described cDNA, LjNOD16, which encodes the L. japonicus late nodulin Nlj16. Nlj16 or highly related amino acid sequences are shown to constitute C-terminal domains of LjPLPs and are suggested to function as specific plasma membrane targeting modules. The expression patterns of one member of this gene family (LjPLP-IV) revealed that LjNOD16 mRNA synthesis in nodules is the result of the transcriptional activity of a nodule specific promoter located in an intron of the LjPLP-IV gene. This intron-borne bidirectional promoter also generates nodule-specific antisense transcripts derived from the N-terminal PITP domain coding region of the LjPLP-IV gene. We propose that Nlj16 protein synthesis and LjPLP-IV antisense transcript generation are components of an elaborate mechanism designed to control LjPLP synthesis and/or functioning in nodules. PMID- 11402167 TI - Molecular characterization of functional domains in the protein kinase SOS2 that is required for plant salt tolerance. AB - The SOS3 (for SALT OVERLY SENSITIVE3) calcium binding protein and SOS2 protein kinase are required for sodium and potassium ion homeostasis and salt tolerance in Arabidopsis. We have shown previously that SOS3 interacts with and activates the SOS2 protein kinase. We report here the identification of a SOS3 binding motif in SOS2 that also serves as the kinase autoinhibitory domain. Yeast two hybrid assays as well as in vitro binding assays revealed a 21-amino acid motif in the regulatory domain of SOS2 that is necessary and sufficient for interaction with SOS3. Database searches revealed a large family of SOS2-like protein kinases containing such a SOS3 binding motif. Using a yeast two-hybrid system, we show that these SOS2-like kinases interact with members of the SOS3 family of calcium binding proteins. Two-hybrid assays also revealed interaction between the N terminal kinase domain and the C-terminal regulatory domain within SOS2, suggesting that the regulatory domain may inhibit kinase activity by blocking substrate access to the catalytic site. Removal of the regulatory domain of SOS2, including the SOS3 binding motif, resulted in constitutive activation of the protein kinase, indicating that the SOS3 binding motif can serve as a kinase autoinhibitory domain. Constitutively active SOS2 that is SOS3 independent also was produced by changing Thr(168) to Asp in the activation loop of the SOS2 kinase domain. Combining the Thr(168)-to-Asp mutation with the autoinhibitory domain deletion created a superactive SOS2 kinase. These results provide insights into regulation of the kinase activities of SOS2 and the SOS2 family of protein kinases. PMID- 11402168 TI - Production of 6-methylsalicylic acid by expression of a fungal polyketide synthase activates disease resistance in tobacco. AB - Salicylic acid (SA) has been shown to act as a signal molecule that is produced by many plants subsequent to the recognition of potentially pathogenic microbes. Increases in levels of SA often trigger the activation of plant defenses and can result in increased resistance to subsequent challenge by pathogens. We observed that the polyketide 6-methylsalicylic acid (6-MeSA), a compound that apparently is not endogenous to tobacco, can mimic SA. Tobacco leaves treated with 6-MeSA show enhanced accumulation of the pathogenesis-related (PR) proteins PR1, beta 1,3-glucanase, and chitinase and also develop increased resistance to tobacco mosaic virus. We transformed tobacco with 6msas, the 6-methylsalicylic acid synthase (6MSAS) gene from Penicillium patulum, to generate plants that constitutively accumulate 6-MeSA. Analysis of primary transformants and the first generation progeny of 6MSAS tobacco revealed that plants can be engineered to accumulate significant amounts of 6-MeSA as a conjugate. Levels of total 6-MeSA increased with plant age. Increased 6-MeSA accumulation correlated with increased levels of PR1 and chitinase proteins and resulted in enhanced resistance of NN genotype 6MSAS tobacco to tobacco mosaic virus. Our results demonstrate that a multistep biosynthetic pathway can be engineered into plants using a single fungal polyketide synthase gene. The functional expression of 6msas can be used to activate disease resistance pathways that normally are induced by SA. PMID- 11402169 TI - A clock- and light-regulated gene that links the circadian oscillator to LHCB gene expression. AB - We have identified and characterized a novel tobacco gene, called ZGT (from the Chinese phrase zhong guang tiaokong, or clock and light controlled), that is regulated by the circadian clock and light. ZGT transcripts have alternate forms that are differentially expressed in different tissues. ZGT is expressed rhythmically in light/dark cycles and in constant light. Constitutive expression of ZGT sustains the expression of the clock-controlled LHCB1*1 gene in constant darkness, when it would normally dampen, but does not affect LHCB1*1 expression in constant light. ZGT expression is induced rapidly by light, and overexpression of ZGT increases the sensitivity of the circadian oscillator to brief light pulses. The ZGT promoter includes a G-box motif that is found in many light regulated promoters in plants and is the same as the E box described for rhythmically regulated promoters of animal circadian clock genes. The ZGT promoter also includes "evening element" motifs that are correlated with circadian control of plant genes. We postulate that light- and clock-regulated expression of ZGT acts as a coupling agent between the central circadian oscillator and rhythmic LHCB1*1 expression and that it may function as a component in plant phototransduction pathways. PMID- 11402170 TI - FPA, a gene involved in floral induction in Arabidopsis, encodes a protein containing RNA-recognition motifs. AB - FPA is a gene that regulates flowering time in Arabidopsis via a pathway that is independent of daylength (the autonomous pathway). Mutations in FPA result in extremely delayed flowering. FPA was identified by means of positional cloning. The predicted FPA protein contains three RNA recognition motifs in the N-terminal region. FPA is expressed most strongly in developing tissues, similar to the expression of FCA and LUMINIDEPENDENS, two components of the autonomous pathway previously identified. Overexpression of FPA in Arabidopsis causes early flowering in noninductive short days and creates plants that exhibit a more day neutral flowering behavior. PMID- 11402171 TI - Proliferating cell nuclear antigen transcription is repressed through an E2F consensus element and activated by geminivirus infection in mature leaves. AB - The geminivirus tomato golden mosaic virus (TGMV) amplifies its DNA genome in differentiated plant cells that lack detectable levels of DNA replication enzymes. Earlier studies showed that TGMV induces the accumulation of proliferating cell nuclear antigen (PCNA), the processivity factor for DNA polymerase delta, in mature cells of Nicotiana benthamiana. We sought to determine if PCNA protein accumulation reflects transcriptional activation of the host gene. RNA gel blot analysis detected an approximately 1200-nucleotide PCNA transcript in young leaves. The same RNA was found in mature leaves of infected but not healthy plants. Reporter gene analysis showed that a 633-bp promoter fragment of the N. benthamiana PCNA gene supports high levels of expression in cultured cells and in young but not mature leaves of healthy transgenic plants. In contrast, PCNA promoter activity was detected in both young and mature leaves of TGMV-infected plants. Developmental studies established a strong relationship between symptom severity, viral DNA accumulation, PCNA promoter activity, and endogenous PCNA mRNA levels. Mutation of an E2F consensus element in the PCNA promoter had no effect on its activity in young leaves but increased transcription in healthy mature leaves. Unlike the wild-type PCNA promoter, TGMV infection had no detectable effect on the activity of the mutant E2F promoter. Together, these results demonstrate that geminivirus infection induces the accumulation of a host replication factor by activating transcription of its gene in mature tissues, most likely by overcoming E2F-mediated repression. PMID- 11402172 TI - Internal aluminum block of plant inward K(+) channels. AB - Aluminum (Al) inhibits inward K(+) channels (K(in)) in both root hair and guard cells, which accounts for at least part of the Al toxicity in plants. To understand the mechanism of Al-induced K(in) inhibition, we performed patch clamp analyses on K(in) in guard cells and on KAT1 channels expressed in Xenopus oocytes. Our results show that Al inhibits plant K(in) by blocking the channels at the cytoplasmic side of the plasma membrane. In guard cells, single-channel recording revealed that Al inhibition of K(in) occurred only upon internal exposure. Using both "giant patch" recording and single-channel analyses, we found that Al reduced KAT1 open probability and changed its activation kinetics through an internal membrane-delimited mechanism. We also provide evidence that a Ca(2)+ channel-like pathway that is sensitive to antagonists verapamil and La(3)+ mediates Al entry across the plasma membrane. We conclude that Al enters plant cells through a Ca(2)+ channel-like pathway and inhibits K(+) uptake by internally blocking K(in). PMID- 11402174 TI - Plant physiology's best paper award 2000. PMID- 11402173 TI - Directed proteomics identifies a plant-specific protein rapidly phosphorylated in response to bacterial and fungal elicitors. AB - The perception of microbial signal molecules is part of the strategy evolved by plants to survive attacks by potential pathogens. To gain a more complete understanding of the early signaling events involved in these responses, we used radioactive orthophosphate to pulse-label suspension-cultured cells of Arabidopsis in conjunction with two-dimensional gel electrophoresis and mass spectrometry to identify proteins that are phosphorylated rapidly in response to bacterial and fungal elicitors. One of these proteins, AtPhos43, and related proteins in tomato and rice, are phosphorylated within minutes after treatment with flagellin or chitin fragments. By measuring (32)P incorporation into AtPhos43 immunoprecipitated from extracts of elicitor-treated hormone and defense response mutants, we found that phosphorylation of AtPhos43 after flagellin treatment but not chitin treatment is dependent on FLS2, a receptor-like kinase involved in flagellin perception. Induction by both elicitors is not dependent on salicylic acid or EDS1, a putative lipase involved in defense signaling. PMID- 11402175 TI - Playing with arabidopsis. PMID- 11402176 TI - A superfamily of proteins with novel cysteine-rich repeats. PMID- 11402177 TI - Fibers. A model for studying cell differentiation, cell elongation, and cell wall biosynthesis. PMID- 11402178 TI - High-throughput screening for induced point mutations. PMID- 11402179 TI - Flavonoid biosynthesis. A colorful model for genetics, biochemistry, cell biology, and biotechnology. PMID- 11402180 TI - Cutting loose. Abscission and dehiscence in Arabidopsis. PMID- 11402181 TI - Challenges and prospects of plant proteomics. PMID- 11402182 TI - Plant cytokinesis. Exploring the links. PMID- 11402183 TI - beta-Aminobutyric acid-induced protection of Arabidopsis against the necrotrophic fungus Botrytis cinerea. AB - The non-protein amino acid beta-aminobutyric acid (BABA) protects numerous plants against various pathogens. Protection of Arabidopsis plants against virulent pathogens involves the potentiation of pathogen-specific defense responses. To extend the analysis of the mode of action of BABA to necrotrophs we evaluated the effect of this chemical on Arabidopsis plants infected with the gray mold fungus Botrytis cinerea. BABA-treated Arabidopsis were found to be less sensitive to two different strains of this pathogen. BABA protected mutants defective in the jasmonate and ethylene pathways, but was inactive in plants impaired in the systemic acquired resistance transduction pathway. Treatments with benzo-(1,2,3) thiadiazole-7-carbothioic acid S-methyl ester, a functional analog of salicylic acid (SA), also markedly reduced the level of infection. Moreover, BABA potentiated mRNA accumulation of the SA-associated PR-1, but not the jasmonate/ethylene-dependent PDF1.2 gene. Thus, besides jasmonate/ethylene dependent defense responses, SA-dependent signaling also contributes to restrict B. cinerea infection in Arabidopsis. Our results also suggest that SA-dependent signaling is down-regulated after infection by B. cinerea. The observed up regulation of the PDF1.2 gene in mutants defective in the SA-dependent signaling pathway points to a cross-talk between SA- and jasmonate/ethylene-dependent signaling pathways during pathogen ingress. PMID- 11402184 TI - Flavonoids act as negative regulators of auxin transport in vivo in arabidopsis. AB - Polar transport of the plant hormone auxin controls many aspects of plant growth and development. A number of synthetic compounds have been shown to block the process of auxin transport by inhibition of the auxin efflux carrier complex. These synthetic auxin transport inhibitors may act by mimicking endogenous molecules. Flavonoids, a class of secondary plant metabolic compounds, have been suggested to be auxin transport inhibitors based on their in vitro activity. The hypothesis that flavonoids regulate auxin transport in vivo was tested in Arabidopsis by comparing wild-type (WT) and transparent testa (tt4) plants with a mutation in the gene encoding the first enzyme in flavonoid biosynthesis, chalcone synthase. In a comparison between tt4 and WT plants, phenotypic differences were observed, including three times as many secondary inflorescence stems, reduced plant height, decreased stem diameter, and increased secondary root development. Growth of WT Arabidopsis plants on naringenin, a biosynthetic precursor to those flavonoids with auxin transport inhibitor activity in vitro, leads to a reduction in root growth and gravitropism, similar to the effects of synthetic auxin transport inhibitors. Analyses of auxin transport in the inflorescence and hypocotyl of independent tt4 alleles indicate that auxin transport is elevated in plants with a tt4 mutation. In hypocotyls of tt4, this elevated transport is reversed when flavonoids are synthesized by growth of plants on the flavonoid precursor, naringenin. These results are consistent with a role for flavonoids as endogenous regulators of auxin transport. PMID- 11402185 TI - Flavonoid accumulation patterns of transparent testa mutants of arabidopsis. AB - Flavonoids have been implicated in the regulation of auxin movements in Arabidopsis. To understand when and where flavonoids may be acting to control auxin movement, the flavonoid accumulation pattern was examined in young seedlings and mature tissues of wild-type Arabidopsis. Using a variety of biochemical and visualization techniques, flavonoid accumulation in mature plants was localized in cauline leaves, pollen, stigmata, and floral primordia, and in the stems of young, actively growing inflorescences. In young Landsberg erecta seedlings, aglycone flavonols accumulated developmentally in three regions, the cotyledonary node, the hypocotyl-root transition zone, and the root tip. Aglycone flavonols accumulated at the hypocotyl-root transition zone in a developmental and tissue-specific manner with kaempferol in the epidermis and quercetin in the cortex. Quercetin localized subcellularly in the nuclear region, plasma membrane, and endomembrane system, whereas kaempferol localized in the nuclear region and plasma membrane. The flavonoid accumulation pattern was also examined in transparent testa mutants blocked at different steps in the flavonoid biosynthesis pathway. The transparent testa mutants were shown to have precursor accumulation patterns similar to those of end product flavonoids in wild-type Landsberg erecta, suggesting that synthesis and end product accumulation occur in the same cells. PMID- 11402186 TI - Alteration of auxin polar transport in the Arabidopsis ifl1 mutants. AB - The INTERFASCICULAR FIBERLESS/REVOLUTA (IFL1/REV) gene is essential for the normal differentiation of interfascicular fibers and secondary xylem in the inflorescence stems of Arabidopsis. It has been proposed that IFL1/REV influences auxin polar flow or the transduction of auxin signal, which is required for fiber and vascular differentiation. Assay of auxin polar transport showed that the ifl1 mutations dramatically reduced auxin polar flow along the inflorescence stems and in the hypocotyls. The null mutant allele ifl1-2 was accompanied by a significant decrease in the expression level of two putative auxin efflux carriers. The ifl1 mutants remained sensitive to auxin and an auxin transport inhibitor. The ifl1-2 mutant exhibited visible phenotypes associated with defects in auxin polar transport such as pin-like inflorescence, reduced numbers of cauline branches, reduced numbers of secondary rosette inflorescence, and dark green leaves with delayed senescence. The visible phenotypes displayed by the ifl1 mutants could be mimicked by treatment of wild-type plants with an auxin polar transport inhibitor. In addition, the auxin polar transport inhibitor altered the normal differentiation of interfascicular fibers in the inflorescence stems of wild-type Arabidopsis. Taken together, these results suggest a correlation between the reduced auxin polar transport and the alteration of cell differentiation and morphology in the ifl1 mutants. PMID- 11402187 TI - The biological functions of glutathione revisited in arabidopsis transgenic plants with altered glutathione levels. AB - A functional analysis of the role of glutathione in protecting plants from environmental stress was undertaken by studying Arabidopsis that had been genetically modified to have altered glutathione levels. The steady-state glutathione concentration in Arabidopsis plants was modified by expressing the cDNA for gamma-glutamyl-cysteine synthetase (GSH1) in both the sense and antisense orientation. The resulting plants had glutathione levels that ranged between 3% and 200% of the level in wild-type plants. Arabidopsis plants with low glutathione levels were hypersensitive to Cd due to the limited capacity of these plants to make phytochelatins. Plants with the lowest levels of reduced glutathione (10% of wild type) were sensitive to as little as 5 microM Cd, whereas those with 50% wild-type levels required higher Cd concentrations to inhibit growth. Elevating glutathione levels did not increase metal resistance. It is interesting that the plants with low glutathione levels were also less able to accumulate anthocyanins supporting a role for glutathione S-transferases for anthocyanin formation or for the vacuolar localization and therefore accumulation of these compounds. Plants with less than 5% of wild-type glutathione levels were smaller and more sensitive to environmental stress but otherwise grew normally. PMID- 11402188 TI - AtCSLD3, a cellulose synthase-like gene important for root hair growth in arabidopsis. AB - A member of the cellulose synthase-like (subfamily D) gene family of Arabidopsis, AtCSLD3, has been identified by T-DNA tagging. The analysis of the corresponding mutant, csld3-1, showed that the AtCSLD3 gene plays a role in root hair growth in plants. Root hairs grow in phases: First a bulge is formed and then the root hair elongates by polarized growth, the so-called "tip growth." In the mutant, root hairs were initiated at the correct position and grew into a bulge, but their elongation was severely reduced. The tips of the csld3-1 root hairs easily leaked cytoplasm, indicating that the tensile strength of the cell wall had changed at the site of the tip. Based on the mutant phenotype and the functional conservation between CSLD3 and the genuine cellulose synthase proteins, we hypothesized that the CSLD3 protein is essential for the synthesis of polymers for the fast-growing primary cell wall at the root hair tip. The distinct mutant phenotype and the ubiquitous expression pattern indicate that the CSLD3 gene product is only limiting at the zone of the root hair tip, suggesting particular physical properties of the cell wall at this specific site of the root hair cell. PMID- 11402189 TI - shygrl1 is a mutant affected in multiple aspects of photomorphogenesis. AB - We have used a counter-selection strategy based on aberrant phytochrome regulation of an Lhcb gene to isolate an Arabidopsis mutant designated shygrl1 (shg1). shg1 seedlings have reduced phytochrome-mediated induction of the Lhcb gene family, but normal phytochrome-mediated induction of several other genes, including the rbcS1a gene. Additional phenotypes observed in shg1 plants include reduced chlorophyll in leaves and additional photomorphogenic abnormalities when the seedlings are grown on medium containing sucrose. Mutations in the TATA proximal region of the Lhcb1*3 promoter that are known to be important for phytochrome regulation affected reporter gene expression in a manner similar to the shg1 mutation. Our results are consistent with the possibility that the mutation either leads to defective chloroplast development or to aberrant phytochrome regulation. They also add to the evidence of complex interactions between light- and sucrose-regulated pathways. PMID- 11402190 TI - The mitochondrial isovaleryl-coenzyme a dehydrogenase of arabidopsis oxidizes intermediates of leucine and valine catabolism. AB - We recently identified a cDNA encoding a putative isovaleryl-coenzyme A (CoA) dehydrogenase in Arabidopsis (AtIVD). In animals, this homotetrameric enzyme is located in mitochondria and catalyzes the conversion of isovaleryl-CoA to 3 methylcrotonyl-CoA as an intermediate step in the leucine (Leu) catabolic pathway. Expression of AtIVD:smGFP4 fusion proteins in tobacco (Nicotiana tabacum) protoplasts and biochemical studies now demonstrate the in vivo import of the plant isovaleryl-CoA dehydrogenase (IVD) into mitochondria and the enzyme in the matrix of these organelles. Two-dimensional separation of mitochondrial proteins by blue native and SDS-PAGE and size determination of the native and overexpressed proteins suggest homodimers to be the dominant form of the plant IVD. Northern-blot hybridization and studies in transgenic Arabidopsis plants expressing Ativd promoter:gus constructs reveal strong expression of this gene in seedlings and young plants grown in the absence of sucrose, whereas promoter activity in almost all tissues is strongly inhibited by exogeneously added sucrose. Substrate specificity tests with AtIVD expressed in Escherichia coli indicate a strong preference toward isovaleryl-CoA but surprisingly also show considerable activity with isobutyryl-CoA. This strongly indicates a commitment of the enzyme in Leu catabolism, but the activity observed with isobutyryl-CoA also suggests a parallel involvement of the enzyme in the dehydrogenation of intermediates of the valine degradation pathway. Such a dual activity has not been observed with the animal IVD and may suggest a novel connection of the Leu and valine catabolism in plants. PMID- 11402191 TI - Transgene expression patterns indicate that spaceflight affects stress signal perception and transduction in arabidopsis. AB - The use of plants as integral components of life support systems remains a cornerstone of strategies for long-term human habitation of space and extraterrestrial colonization. Spaceflight experiments over the past few decades have refined the hardware required to grow plants in low-earth orbit and have illuminated fundamental issues regarding spaceflight effects on plant growth and development. Potential incipient hypoxia, resulting from the lack of convection driven gas movement, has emerged as a possible major impact of microgravity. We developed transgenic Arabidopsis containing the alcohol dehydrogenase (Adh) gene promoter linked to the beta-glucuronidase (GUS) reporter gene to address specifically the possibility that spaceflight induces the plant hypoxia response and to assess whether any spaceflight response was similar to control terrestrial hypoxia-induced gene expression patterns. The staining patterns resulting from a 5-d mission on the orbiter Columbia during mission STS-93 indicate that the Adh/GUS reporter gene was activated in roots during the flight. However, the patterns of expression were not identical to terrestrial control inductions. Moreover, although terrestrial hypoxia induces Adh/GUS expression in the shoot apex, no apex staining was observed in the spaceflight plants. This indicates that either the normal hypoxia response signaling is impaired in spaceflight or that spaceflight inappropriately induces Adh/GUS activity for reasons other than hypoxia. PMID- 11402192 TI - Male meiotic spindle lengths in normal and mutant arabidopsis cells. AB - Spindle elongation is crucial to normal chromosome separation in eukaryotes; in particular, it is required for or associated with the extension of distance between spindle poles and the further moving apart of the already separated chromosomes. However, little is known about the relationship between spindle elongation and the status of chromosome separation, and it is unknown whether spindle elongation in different organisms shares any quantitative feature. The Arabidopsis ask1-1 mutant might be a unique material for addressing these questions because it appears to have functional spindles, but a severe defect in homolog separation at male anaphase I (M. Yang, Y. Hu, M. Lodhi, W.R. McCombie, H Ma [1999] Proc Natl Acad Sci USA 96: 11416-11421). We have characterized male meiotic spindle lengths in wild-type and the ask1-1 mutant plants. We observed that during meiosis I some ask1-1 cells had spindles that were similar in length to fully elongated normal spindles, but the chromosomes in these cells did not show appreciable movement from the equator. Furthermore, greater movement of chromosomes from the equator was usually found in the ask1-1 cells that had longer than normal spindles. These results suggest that additional elongation of ask1-1 spindles occurred; one possible reason for the extra-long spindles may be that it is a consequence of chromosome non-separation. We also found that normal and ask1-1 spindle lengths are clustered at discrete values, and their differences are of multiples of 0.7 microm. A search of the literature revealed that in each of several organisms, spindle lengths also differ by multiples of 0.7 microm. These findings strongly suggest that the spindle elongates in response to status of chromosome separation, and perhaps there are conserved mechanisms controlling the extent of spindle elongation. PMID- 11402193 TI - The three-dimensional structure of cystathionine beta-lyase from Arabidopsis and its substrate specificity. AB - The pyridoxal 5'-phosphate-dependent enzyme cystathionine beta-lyase (CBL) catalyzes the penultimate step in the de novo biosynthesis of Met in microbes and plants. Absence of CBL in higher organisms makes it an important target for the development of antibiotics and herbicides. The three-dimensional structure of cystathionine beta-lyase from Arabidopsis was determined by Patterson search techniques, using the structure of tobacco (Nicotiana tabacum) cystathionine gamma-synthase as starting point. At a resolution of 2.3 A, the model was refined to a final crystallographic R-factor of 24.9%. The overall structure is very similar to other pyridoxal 5'-phosphate-dependent enzymes of the gamma-family. Exchange of a few critical residues within the active site causes the different substrate preferences between Escherichia coli and Arabidopsis CBL. Loss of interactions at the alpha-carboxyl site is the reason for the poorer substrate binding of Arabidopsis CBL. In addition, the binding pocket of Arabidopsis CBL is larger than that of E. coli CBL, explaining the similar binding of L cystathionine and L-djenkolate in Arabidopsis CBL in contrast to E. coli CBL, where the substrate binding site is optimized for the natural substrate cystathionine. PMID- 11402194 TI - The arabidopsis ATHB-8 HD-zip protein acts as a differentiation-promoting transcription factor of the vascular meristems. AB - ATHB-8, -9, -14, -15, and IFL1/REV are members of a small homeodomain-leucine zipper family whose genes are characterized by expression in the vascular tissue. ATHB-8, a gene positively regulated by auxin (Baima et al., 1995), is considered an early marker of the procambial cells and of the cambium during vascular regeneration after wounding. Here, we demonstrate that although the formation of the vascular system is not affected in athb8 mutants, ectopic expression of ATHB 8 in Arabidopsis plants increased the production of xylem tissue. In particular, a careful anatomical analysis of the transgenic plants indicated that the overexpression of ATHB-8 promotes vascular cell differentiation. First, the procambial cells differentiated precociously into primary xylem. In addition, interfascicular cells also differentiated precociously into fibers. Finally, the transition to secondary growth, mainly producing xylem, was anticipated in transgenic inflorescence stems compared with controls. The stimulation of primary and secondary vascular cell differentiation resulted in complex modifications of the growth and development of the ATHB-8 transgenic plants. Taken together, these results are consistent with the hypothesis that ATHB-8 is a positive regulator of proliferation and differentiation, and participates in a positive feedback loop in which auxin signaling induces the expression of ATHB-8, which in turn positively modulates the activity of procambial and cambial cells to differentiate. PMID- 11402195 TI - The heme-oxygenase family required for phytochrome chromophore biosynthesis is necessary for proper photomorphogenesis in higher plants. AB - The committed step in the biosynthesis of the phytochrome chromophore phytochromobilin involves the oxidative cleavage of heme by a heme oxygenase (HO) to form biliverdin IXalpha. Through positional cloning of the photomorphogenic mutant hy1, the Arabidopsis HO (designated AtHO1) responsible for much of phytochromobilin synthesis recently was identified. Using the AtHO1 sequence, we identified families of HO genes in a number of plants that cluster into two subfamilies (HO1- and HO2-like). The tomato (Lycopersicon esculentum) yg-2 and Nicotiana plumbaginifolia pew1 photomorphogenic mutants are defective in specific HO genes. Phenotypic analysis of a T-DNA insertion mutant of Arabidopsis HO2 revealed that the second HO subfamily also contributes to phytochromobilin synthesis. Homozygous ho2-1 plants show decreased chlorophyll accumulation, reduced growth rate, accelerated flowering time, and reduced de-etiolation. A mixture of apo- and holo-phyA was detected in etiolated ho2-1 seedlings, suggesting that phytochromobilin is limiting in this mutant, even in the presence of functional AtHO1. The patterns of Arabidopsis HO1 and HO2 expression suggest that the products of both genes overlap temporally and spatially. Taken together, the family of HOs is important for phytochrome-mediated development in a number of plants and that each family member may uniquely contribute to the phytochromobilin pool needed to assemble holo-phytochromes. PMID- 11402196 TI - The Rop GTPase switch controls multiple developmental processes in Arabidopsis. AB - G proteins are universal molecular switches in eukaryotic signal transduction. The Arabidopsis genome sequence reveals no RAS small GTPase and only one or a few heterotrimeric G proteins, two predominant classes of signaling G proteins found in animals. In contrast, Arabidopsis possesses a unique family of 11 Rop GTPases that belong to the Rho family of small GTPases. Previous studies indicate that Rop controls actin-dependent pollen tube growth and H(2)O(2)-dependent defense responses. In this study, we tested the hypothesis that the Rop GTPase acts as a versatile molecular switch in signaling to multiple developmental processes in Arabidopsis. Immunolocalization using a general antibody against the Rop family proteins revealed a ubiquitous distribution of Rop proteins in all vegetative and reproductive tissues and cells in Arabidopsis. The cauliflower mosaic virus 35S promoter-directed expression of constitutively active GTP-bound rop2 (CA-rop2) and dominant negative GDP-bound rop2 (DN-rop2) mutant genes impacted many aspects of plant growth and development, including embryo development, seed dormancy, seedling development, lateral root initiation, morphogenesis of lateral organs in the shoot, shoot apical dominance and growth, phyllotaxis, and lateral organ orientation. The rop2 transgenic plants also displayed altered responses to the exogenous application of several hormones, such as abscisic acid-mediated seed dormancy, auxin-dependent lateral shoot initiation, and brassinolide-mediated hypocotyl elongation. CA-rop2 and DN-rop2 expression had opposite effects on most of the affected processes, supporting a direct signaling role for Rop in regulating these processes. Based on these observations and previous results, we propose that Rop2 and other members of the Rop family participate in multiple distinct signaling pathways that control plant growth, development, and responses to the environment. PMID- 11402197 TI - Pollen germinates precociously in the anthers of raring-to-go, an Arabidopsis gametophytic mutant. AB - Pollen hydration is usually tightly regulated and occurs in vivo only when desiccated pollen grains acquire water from the female, thus enabling pollen tube growth. Pollen tubes are easily visualized by staining with decolorized aniline blue, a stain specific for callose. We identified a mutant, raring-to-go, in which pollen grains stained for callose before anther dehiscence. When raring-to go plants are transferred to high humidity, pollen tubes dramatically elongate within the anther. As early as the bicellular stage, affected pollen grains in raring-to-go plants acquire or retain water within the anther, and precociously germinate. Thus, the requirement for contact with the female is circumvented. We used pollen tetrad analysis to show that raring-to-go is a gametophytic mutation, to our knowledge the first gametophytic mutation in Arabidopsis that affects early events in the pollination pathway. To aid in identifying raring-to-go alleles, we devised a new technique for screening pollen in bulk with decolorized aniline blue. We screened a new M(1) mutagenized population and identified several additional mutants with a raring-to-go-like phenotype, demonstrating the usefulness of this technique. Further, we isolated other mutants (gift-wrapped pollen, polka dot pollen, and emotionally fragile pollen) with unexpected patterns of callose staining. We suggest that raring-to-go and these other mutants may help dissect components of the pathway that regulates pollen hydration and pollen tube growth. PMID- 11402198 TI - Inventory of the superfamily of P-type ion pumps in Arabidopsis. AB - A total of 45 genes encoding for P-type ATPases have been identified in the complete genome sequence of Arabidopsis. Thus, this plant harbors a primary transport capability not seen in any other eukaryotic organism sequenced so far. The sequences group in all five subfamilies of P-type ATPases. The most prominent subfamilies are P(1B) ATPases (heavy metal pumps; seven members), P(2A) and P(2B) ATPases (Ca(2+) pumps; 14 in total), P(3A) ATPases (plasma membrane H(+) pumps; 12 members including a truncated pump, which might represent a pseudogene or an ATPase-like protein with an alternative function), and P(4) ATPases (12 members). P(4) ATPases have been implicated in aminophosholipid flipping but it is not known whether this is a direct or an indirect effect of pump activity. Despite this apparent plethora of pumps, Arabidopsis appears to be lacking Na(+) pumps and secretory pathway (PMR1-like) Ca(2+)-ATPases. A cluster of Arabidopsis heavy metal pumps resembles bacterial Zn(2+)/Co(2+)/Cd(2+)/Pb(2+) transporters. Two members of the cluster have extended C termini containing putative heavy metal binding motifs. The complete inventory of P-type ATPases in Arabidopsis is an important starting point for reverse genetic and physiological approaches aiming at elucidating the biological significance of these pumps. PMID- 11402199 TI - Networking senescence-regulating pathways by using Arabidopsis enhancer trap lines. AB - The last phase of leaf development, generally referred to as leaf senescence, is an integral part of plant development that involves massive programmed cell death. Due to a sharp decline of photosynthetic capacity in a leaf, senescence limits crop yield and forest plant biomass production. However, the biochemical components and regulatory mechanisms underlying leaf senescence are poorly characterized. Although several approaches such as differential cDNA screening, differential display, and cDNA subtraction have been employed to isolate senescence-associated genes (SAGs), only a limited number of SAGs have been identified, and information regarding the regulation of these genes is fragmentary. Here we report on the utilization of enhancer trap approach toward the identification and analysis of SAGs. We have developed a sensitive large scale screening method and have screened 1,300 Arabidopsis enhancer trap lines and have identified 147 lines in which the reporter gene GUS (beta-glucuronidase) is expressed in senescing leaves but not in non-senescing ones. We have systematically analyzed the regulation of beta-glucuronidase expression in 125 lines (genetically, each contains single T-DNA insertion) by six senescence promoting factors, namely abscisic acid, ethylene, jasmonic acid, brassinosteroid, darkness, and dehydration. This analysis not only reveals the complexity of the regulatory circuitry but also allows us to postulate the existence of a network of senescence-promoting pathways. We have also cloned three SAGs from randomly selected enhancer trap lines, demonstrating that reporter expression pattern reflects the expression pattern of the endogenous gene. PMID- 11402200 TI - The Arabidopsis embryo mutant schlepperless has a defect in the chaperonin 60alpha gene. AB - We identified a T-DNA-generated mutation in the chaperonin-60alpha gene of Arabidopsis that produces a defect in embryo development. The mutation, termed schlepperless (slp), causes retardation of embryo development before the heart stage, even though embryo morphology remains normal. Beyond the heart stage, the slp mutation results in defective embryos with highly reduced cotyledons. slp embryos exhibit a normal apical-basal pattern and radial tissue organization, but they are morphologically retarded. Even though slp embryos are competent to transcribe two late-maturation gene markers, this competence is acquired more slowly as compared with wild-type embryos. slp embryos also exhibit a defect in plastid development-they remain white during maturation in planta and in culture. Hence, the overall developmental phenotype of the slp mutant reflects a lesion in the chloroplast that affects embryo development. The slp phenotype highlights the importance of the chaperonin-60alpha protein for chloroplast development and subsequently for the proper development of the plant embryo and seedling. PMID- 11402201 TI - Nitrite reductase gene enrichment improves assimilation of NO(2) in Arabidopsis. AB - Transgenic plants of Arabidopsis bearing the spinach (Spinacia oleracea) nitrite reductase (NiR, EC 1.7.7.1) gene that catalyzes the six-electron reduction of nitrite to ammonium in the second step of the nitrate assimilation pathway were produced by use of the cauliflower mosaic virus 35S promoter and nopaline synthase terminator. Integration of the gene was confirmed by a genomic polymerase chain reaction (PCR) and Southern-blot analysis; its expression by a reverse transcriptase-PCR and two-dimensional polyacrylamide gel electrophoresis western-blot analysis; total (spinach + Arabidopsis) NiR mRNA content by a competitive reverse transcriptase-PCR; localization of NiR activity (NiRA) in the chloroplast by fractionation analysis; and NO(2) assimilation by analysis of the reduced nitrogen derived from NO(2) (NO(2)-RN). Twelve independent transgenic plant lines were characterized in depth. Three positive correlations were found for NiR gene expression; between the total NiR mRNA and total NiR protein contents (r = 0.74), between the total NiR protein and NiRA (r = 0.71), and between NiRA and NO(2)-RN (r = 0.65). Of these twelve lines, four had significantly higher NiRA than the wild-type control (P < 0.01), and three had significantly higher NO(2)-RN (P < 0.01). Each of the latter three had one to two copies of spinach NiR cDNA per haploid genome. The NiR flux control coefficient for NO(2) assimilation was estimated to be about 0.4. A similar value was obtained for an NiR antisense tobacco (Nicotiana tabacum cv Xanthi XHFD8). The flux control coefficients of nitrate reductase and glutamine synthetase were much smaller than this value. Together, these findings indicate that NiR is a controlling enzyme in NO(2) assimilation by plants. PMID- 11402202 TI - Signaling events in the hypoxic induction of alcohol dehydrogenase gene in Arabidopsis. AB - Expression of the alcohol dehydrogenase gene (ADH) of Arabidopsis is induced during hypoxia. Because many plants increase their ethylene production in response to hypoxic stress, we examined in this report whether ethylene is involved in the hypoxic induction of ADH in Arabidopsis. We found that the hypoxic induction of ADH can be partially inhibited by aminooxy acetic acid, an inhibitor of ethylene biosynthesis. This partial inhibition can be reversed by the addition of 1-aminocyclopropane-1-carboxylic acid, a direct precursor of ethylene. In addition, the hypoxic induction of the ADH gene is also reduced in etr1-1 and ein2-1, two ethylene insensitive mutants in ethylene-signaling pathways, whereas the addition of exogenous ethylene or an increase in cellular ethylene alone does not induce ADH under normoxic conditions. Kinetic analyses of ADH mRNA accumulation indicated that an ethylene signal is required for the induction of ADH during later stages of hypoxia. Therefore, we conclude that ethylene is needed, but not sufficient for, the induction of ADH in Arabidopsis during hypoxia. PMID- 11402203 TI - Transgenic Arabidopsis plants with decreased activity of fructose-6-phosphate,2 kinase/fructose-2,6-bisphosphatase have altered carbon partitioning. AB - The role of fructose-2,6-bisphosphate (Fru-2,6-P(2)) as a regulatory metabolite in photosynthetic carbohydrate metabolism was studied in transgenic Arabidopsis plants with reduced activity of Fru-6-phosphate,2-kinase/Fru-2,6-bisphosphatase. A positive correlation was observed between the Fru-6-phosphate,2-kinase activity and the level of Fru-2,6-P(2) in the leaves. The partitioning of carbon was studied by (14)CO(2) labeling of photosynthetic products. Plant lines with Fru 2,6-P(2) levels down to 5% of the levels observed in wild-type (WT) plants had significantly altered partitioning of carbon between sucrose (Suc) versus starch. The ratio of (14)C incorporated into Suc and starch increased 2- to 3-fold in the plants with low levels of Fru-2,6-P(2) compared with WT. Transgenic plant lines with intermediate levels of Fru-2,6-P(2) compared with WT had a Suc-to-starch labeling ratio similar to the WT. Levels of sugars, starch, and phosphorylated intermediates in leaves were followed during the diurnal cycle. Plants with low levels of Fru-2,6-P(2) in leaves had high levels of Suc, glucose, and Fru and low levels of triose phosphates and glucose-1-P during the light period compared with WT. During the dark period these differences were eliminated. Our data provide direct evidence that Fru-2,6-P(2) affects photosynthetic carbon partitioning in Arabidopsis. Opposed to this, Fru-2,6-P(2) does not contribute significantly to regulation of metabolite levels in darkness. PMID- 11402204 TI - Rapid accumulation of phosphatidylinositol 4,5-bisphosphate and inositol 1,4,5 trisphosphate correlates with calcium mobilization in salt-stressed arabidopsis. AB - The phosphoinositide phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P(2)] is a key signaling molecule in animal cells. It can be hydrolyzed to release 1,2 diacyglycerol and inositol 1,4,5-trisphosphate (IP(3)), which in animal cells lead to protein kinase C activation and cellular calcium mobilization, respectively. In addition to its critical roles in constitutive and regulated secretion of proteins, PtdIns(4,5)P(2) binds to proteins that modify cytoskeletal architecture and phospholipid constituents. Herein, we report that Arabidopsis plants grown in liquid media rapidly increase PtdIns(4,5)P(2) synthesis in response to treatment with sodium chloride, potassium chloride, and sorbitol. These results demonstrate that when challenged with salinity and osmotic stress, terrestrial plants respond differently than algae, yeasts, and animal cells that accumulate different species of phosphoinositides. We also show data demonstrating that whole-plant IP(3) levels increase significantly within 1 min of stress initiation, and that IP(3) levels continue to increase for more than 30 min during stress application. Furthermore, using the calcium indicators Fura-2 and Fluo-3 we show that root intracellular calcium concentrations increase in response to stress treatments. Taken together, these results suggest that in response to salt and osmotic stress, Arabidopsis uses a signaling pathway in which a small but significant portion of PtdIns(4,5)P(2) is hydrolyzed to IP(3). The accumulation of IP(3) occurs during a time frame similar to that observed for stress-induced calcium mobilization. These data also suggest that the majority of the PtdIns(4,5)P(2) synthesized in response to salt and osmotic stress may be utilized for cellular signaling events distinct from the canonical IP(3) signaling pathway. PMID- 11402205 TI - Brassinosteroid-6-oxidases from Arabidopsis and tomato catalyze multiple C-6 oxidations in brassinosteroid biosynthesis. AB - Brassinosteroids (BRs) are steroidal plant hormones that are essential for growth and development. It has been proposed that BRs are synthesized via two parallel pathways, the early and late C-6 oxidation pathways according to the C-6 oxidation status. The tomato (Lycopersicon esculentum) Dwarf gene encodes a cytochrome P450 that has been shown to catalyze the C-6 oxidation of 6 deoxocastasterone to castasterone. We isolated an Arabidopsis ortholog (AtBR6ox gene) of the tomato Dwarf gene. The encoded polypeptide has characteristics of P450s and is classified into the CYP85 family. The AtBR6ox and tomato Dwarf gene were expressed in yeast and the ability of the transformed yeast cells to metabolize 6-deoxo-BRs was tested. Metabolites were analyzed by gas chromatography-mass spectrometry. Both enzymes catalyze multiple steps in BR biosynthesis: 6-deoxoteasterone to teasterone, 3-dehydro-6-deoxoteasterone to 3 dehydroteasterone, 6-deoxotyphasterol to typhasterol, and 6-deoxocastasterone to castasterone. Our results indicate that the AtBR6ox gene and the tomato Dwarf gene encode steroid-6-oxidases and that these enzymes have a broad substrate specificity. This suggests that the BR biosynthetic pathway consists of a metabolic grid rather than two separate parallel pathways. PMID- 11402206 TI - Ultraviolet B radiation enhances a phytochrome-B-mediated photomorphogenic response in Arabidopsis. AB - Ultraviolet B radiation (UV-B, 290-315 nm) can cause damage and induce photomorphogenic responses in plants. The mechanisms that mediate the photomorphogenic effects of UV-B are unclear. In etiolated Arabidopsis seedlings, a daily exposure to 2.5 h of UV-B enhanced the cotyledon opening response induced by a subsequent red light (R) pulse. An R pulse alone, 2.5 h of UV-B terminated with a far-red pulse, or 2.5 h of continuous R caused very little cotyledon opening. The enhancing effect of UV-B increased with fluence rate up to approximately 7.58 micromol m(-2) s(-1); at higher fluence rates the response to UV-B was greatly reduced. The phyA, phyA cry1, and cry1 cry2 mutants behaved like the wild type when exposed to UV-B followed by an R pulse. In contrast, phyB, phyB cry1, and phyB phyA mutants failed to open the cotyledons. Thus, phytochrome B was required for the cotyledon opening response to UV-B --> R treatments, whereas phytochrome A and cryptochromes 1 and 2 were not necessary under the conditions of our experiments. The enhancing effect of low doses of UV-B on cotyledon opening in uvr1 uvr2 and uvr1 uvr3 mutants, deficient in DNA repair, was similar to that found in the wild type, suggesting that this effect of UV-B was not elicited by signals derived from UV-B-induced DNA lesions (cyclobutane pyrimidine dimers and 6-4 photoproducts). We conclude that low doses of UV-B, perceived by a receptor system different from phytochromes, cryptochromes, or DNA, enhance a de-etiolation response that is induced by active phytochrome B. PMID- 11402207 TI - Comprehensive expression profile analysis of the Arabidopsis Hsp70 gene family. AB - We isolated cDNA clones for two nuclear-encoded, organellar members of the Arabidopsis hsp70 gene family, mtHsc70-2 (AF217458) and cpHsc70-2 (AF217459). Together with the completion of the genome sequence, the hsp70 family in Arabidopsis consists of 14 members unequally distributed among the five chromosomes. To establish detailed expression data of this gene family, a comprehensive reverse transcriptase-polymerase chain reaction analysis for 11 hsp70s was conducted including analysis of organ-specific and developmental expression and expression in response to temperature extremes. All hsp70s showed 2- to 20-fold induction by heat shock treatment except cpHsc70-1 and mtHsc70-1, which were unchanged or repressed. The expression profiles in response to low temperature treatment were more diverse than those evoked by heat shock treatment. Both mitochondrial and all cytosolic members of the family except Hsp70b were strongly induced by low temperature, whereas endoplasmic reticulum and chloroplast members were not induced or were slightly repressed. Developmentally regulated expression of the heat-inducible Hsp70 in mature dry seed and roots in the absence of temperature stress suggests prominent roles in seed maturation and root growth for this member of the hsp70 family. This reverse transcriptase-polymerase chain reaction analysis establishes the complex differential expression pattern for the hsp70s in Arabidopsis that portends specialized functions even among members localized to the same subcellular compartment. PMID- 11402208 TI - Molecular characterization of At5PTase1, an inositol phosphatase capable of terminating inositol trisphosphate signaling. AB - The inositol triphosphate (IP(3))-signaling pathway has been associated with several developmental and physiological processes in plants, but we currently know little about the regulation of this pathway. Inositol 5' phosphatases (5PTases) are enzymes that remove a 5' phosphate from several potential second messengers, including IP(3). In catalyzing the removal of a 5' phosphate from second messenger substrates, 5PTases can act to terminate signal transduction events. We describe the molecular analysis of At5PTase1, a 5PTase gene from Arabidopsis. When expressed transiently in Arabidopsis leaf tissue or ectopically in transgenic plants, At5PTase1 allowed for the increased hydrolysis of I(1,4,5)P(3) and I(1,3,4,5)P(4) substrates. At5PTase1 did not hydrolyze I(1)P, I(1,4)P(2), or PI(4,5)P(2) substrates. This substrate specificity was similar to that of the human Type I 5PTase. We identified 14 other potential At5PTase genes and constructed an unrooted phylogenetic tree containing putative Arabidopsis, human, and yeast 5PTase proteins. This analysis indicated that the Arabidopsis 5PTases were grouped in two separate branches of the tree. The multiplicity of At5PTases indicates that these enzymes may have different substrate specificities and play different roles in signal termination in Arabidopsis. PMID- 11402209 TI - Genetic control of natural variation in Arabidopsis glucosinolate accumulation. AB - Glucosinolates are biologically active secondary metabolites of the Brassicaceae and related plant families that influence plant/insect interactions. Specific glucosinolates can act as feeding deterrents or stimulants, depending upon the insect species. Hence, natural selection might favor the presence of diverse glucosinolate profiles within a given species. We determined quantitative and qualitative variation in glucosinolates in the leaves and seeds of 39 Arabidopsis ecotypes. We identified 34 different glucosinolates, of which the majority are chain-elongated compounds derived from methionine. Polymorphism at only five loci was sufficient to generate 14 qualitatitvely different leaf glucosinolate profiles. Thus, there appears to be a modular genetic system regulating glucosinolate profiles in Arabidopsis. This system allows the rapid generation of new glucosinolate combinations in response to changing herbivory or other selective pressures. In addition to the qualitative variation in glucosinolate profiles, we found a nearly 20-fold difference in the quantity of total aliphatic glucosinolates and were able to identify a single locus that controls nearly three-quarters of this variation. PMID- 11402210 TI - The enhancement of phototropin-induced phototropic curvature in Arabidopsis occurs via a photoreversible phytochrome A-dependent modulation of auxin responsiveness. AB - The induction of phototropism in etiolated (dark-grown) seedlings exposed to an unidirectional pulse or extended irradiation with low fluence rate blue light (BL) requires the action of the phototropin (nph1) BL receptor. Although cryptochromes and phytochromes are not required for phototropic induction, these photoreceptors do modulate the magnitude of curvature resulting from phototropin activation. Modulatory increases in the magnitude of phototropic curvature have been termed "enhancement." Here, we show that phototropic enhancement is primarily a phytochrome A (phyA)-dependent red/far-red-reversible low fluence response. This phyA-dependent response is genetically separable from the basal phototropin-dependent response, as demonstrated by its retention under extended irradiation conditions in the nph4 mutant background, which normally lacks the basal BL-induced response. It is interesting that the nph4 mutants fail to exhibit the basal phototropin-dependent and phyA-dependent enhancement responses under limiting light conditions. Given that NPH4 encodes a transcriptional activator, auxin response factor 7 (ARF7), we hypothesize that the ultimate target(s) of phyA action during the phototropic enhancement response is a rate limiting ARF-containing transcriptional complex in which the constituent ARFs can vary in identity or activity depending upon the irradiation condition. PMID- 11402211 TI - Proteomic analysis of arabidopsis seed germination and priming. AB - To better understand seed germination, a complex developmental process, we developed a proteome analysis of the model plant Arabidopsis for which complete genome sequence is now available. Among about 1,300 total seed proteins resolved in two-dimensional gels, changes in the abundance (up- and down-regulation) of 74 proteins were observed during germination sensu stricto (i.e. prior to radicle emergence) and the radicle protrusion step. This approach was also used to analyze protein changes occurring during industrial seed pretreatments such as priming that accelerate seed germination and improve seedling uniformity. Several proteins were identified by matrix-assisted laser-desorption ionization time of flight mass spectrometry. Some of them had previously been shown to play a role during germination and/or priming in several plant species, a finding that underlines the usefulness of using Arabidopsis as a model system for molecular analysis of seed quality. Furthermore, the present study, carried out at the protein level, validates previous results obtained at the level of gene expression (e.g. from quantitation of differentially expressed mRNAs or analyses of promoter/reporter constructs). Finally, this approach revealed new proteins associated with the different phases of seed germination and priming. Some of them are involved either in the imbibition process of the seeds (such as an actin isoform or a WD-40 repeat protein) or in the seed dehydration process (e.g. cytosolic glyceraldehyde-3-phosphate dehydrogenase). These facts highlight the power of proteomics to unravel specific features of complex developmental processes such as germination and to detect protein markers that can be used to characterize seed vigor of commercial seed lots and to develop and monitor priming treatments. PMID- 11402212 TI - Jasmonate-dependent induction of indole glucosinolates in Arabidopsis by culture filtrates of the nonspecific pathogen Erwinia carotovora. AB - Elicitors from the plant pathogen Erwinia carotovora trigger coordinate induction of the tryptophan (Trp) biosynthesis pathway and Trp oxidizing genes in Arabidopsis. To elucidate the biological role of such pathogen-induced activation we characterized the production of secondary defense metabolites such as camalexin and indole glucosinolates derived from precursors of this pathway. Elicitor induction was followed by a specific increase in 3 indolylmethylglucosinolate (IGS) content, but only a barely detectable accumulation of the indole-derived phytoalexin camalexin. The response is mediated by jasmonic acid as shown by lack of IGS induction in the jasmonate insensitive mutant coi1-1. In accordance with this, methyl jasmonate was able to trigger IGS accumulation in Arabidopsis. In contrast, ethylene and salicylic acid seem to play a minor role in the response. They did not trigger alterations in IGS levels, and methyl jasmonate- or elicitor-induced IGS accumulation in NahG and ethylene-insensitive ein2-1 mutant plants was similar as in the wild type. The breakdown products of IGS and other glucosinolates were able to inhibit growth of E. carotovora. The results suggest that IGS is of importance in the defense against bacterial pathogens. PMID- 11402213 TI - Seed-specific over-expression of an Arabidopsis cDNA encoding a diacylglycerol acyltransferase enhances seed oil content and seed weight. AB - We recently reported the cloning and characterization of an Arabidopsis (ecotype Columbia) diacylglycerol acyltransferase cDNA (Zou et al., 1999) and found that in Arabidopsis mutant line AS11, an ethyl methanesulfonate-induced mutation at a locus on chromosome II designated as Tag1 consists of a 147-bp insertion in the DNA, which results in a repeat of the 81-bp exon 2 in the Tag1 cDNA. This insertion mutation is correlated with an altered seed fatty acid composition, reduced diacylglycerol acyltransferase (DGAT; EC 2.3.1.20) activity, reduced seed triacylglycerol content, and delayed seed development in the AS11 mutant. The effect of the insertion mutation on microsomal acyl-coenzyme A-dependent DGAT is examined with respect to DGAT activity and its substrate specificity in the AS11 mutant relative to wild type. We demonstrate that transformation of mutant AS11 with a single copy of the wild-type Tag1 DGAT cDNA can complement the fatty acid and reduced oil phenotype of mutant AS11. More importantly, we show for the first time that seed-specific over-expression of the DGAT cDNA in wild-type Arabidopsis enhances oil deposition and average seed weight, which are correlated with DGAT transcript levels. The DGAT activity in developing seed of transgenic lines was enhanced by 10% to 70%. Thus, the current study confirms the important role of DGAT in regulating the quantity of seed triacylglycerols and the sink size in developing seeds. PMID- 11402214 TI - Phosphate availability regulates root system architecture in Arabidopsis. AB - Plant root systems are highly plastic in their development and can adapt their architecture in response to the prevailing environmental conditions. One important parameter is the availability of phosphate, which is highly immobile in soil such that the arrangement of roots within the soil will profoundly affect the ability of the plant to acquire this essential nutrient. Consistent with this, the availability of phosphate was found to have a marked effect on the root system architecture of Arabidopsis. Low phosphate availability favored lateral root growth over primary root growth, through increased lateral root density and length, and reduced primary root growth mediated by reduced cell elongation. The ability of the root system to respond to phosphate availability was found to be independent of sucrose supply and auxin signaling. In contrast, shoot phosphate status was found to influence the root system architecture response to phosphate availability. PMID- 11402215 TI - Induction of ApL3 expression by trehalose complements the starch-deficient Arabidopsis mutant adg2-1 lacking ApL1, the large subunit of ADP-glucose pyrophosphorylase. AB - The disaccharide trehalose has strong effects on plant metabolism and development. In Arabidopsis seedlings, growth on trehalose-containing medium leads to an inhibition of root elongation, an accumulation of starch in the shoots, an increased activity of ADP-Glc pyrophosphorylase (AGPase), and an induction of the expression of the AGPase gene, ApL3 (A. Wingler, T. Fritzius, A. Wiemken, T. Boller, R.A. Aeschbacher [2000] Plant Physiol 124: 105-114). We used Arabidopsis mutants deficient in starch synthesis to examine whether the primary effect of trehalose was to affect carbohydrate allocation by the induction of AGPase in the photosynthetic tissue. In a mutant lacking the large AGPase subunit, ApL1, (adg2-1 mutant) growth on trehalose restored AGPase activity and led to a strong accumulation of starch in the shoots. In contrast, starch synthesis could not be induced in a mutant lacking the small AGPase subunit, ApS, (adg1-1 mutant) or in a mutant lacking plastidic phosphoglucomutase (pgm1-1 mutant). These results indicate that ApL3 can substitute for ApL1 in the AGPase complex. In addition, root elongation in the mutants, especially in the adg1-1 mutant, was partially resistant to trehalose, suggesting that the induction of ApL3 expression and the resulting accumulation of starch in the shoots were partially responsible for the effects of trehalose on the growth of wild-type plants. PMID- 11402216 TI - The TASTY locus on chromosome 1 of Arabidopsis affects feeding of the insect herbivore Trichoplusia ni. AB - The generalist insect herbivore Trichoplusia ni (cabbage looper) readily consumes Arabidopsis and can complete its entire life cycle on this plant. Natural isolates (ecotypes) of Arabidopsis are not equally susceptible to T. ni feeding. While some are hardly touched by T. ni, others are eaten completely to the ground. Comparison of two commonly studied Arabidopsis ecotypes in choice experiments showed that Columbia is considerably more resistant than Landsberg erecta. In no-choice experiments, where larvae were confined on one or the other ecotype, weight gain was more rapid on Landsberg erecta than on Columbia. Genetic mapping of this difference in insect susceptibility using recombinant inbred lines resulted in the discovery of the TASTY locus near 85 cM on chromosome 1 of Arabidopsis. The resistant allele of this locus is in the Columbia ecotype, and an F(1) hybrid has a sensitive phenotype that is similar to that of Landsberg erecta. The TASTY locus is distinct from known genetic differences between Columbia and Landsberg erecta that affect glucosinolate content, trichome density, disease resistance, and flowering time. PMID- 11402217 TI - Novel patterns of ectopic cell plate growth and lipid body distribution in the Arabidopsis gemini pollen1 mutant. AB - The nature of aberrant gametophytic cell divisions and altered pollen cell fate in the gemini pollen1 (gem1) mutant was investigated through ultrastructural analysis. The earliest noticeable defect in gem1 was the appearance of extended membrane profiles at the early bicellular stage. These were replaced by ectopic internal walls, which divided the cytoplasm into twin or multiple cell compartments. Complete or partial internal walls were callosic with highly complex profiles, indicating failed guidance or deregulated cell plate growth. Extended membrane profiles and delayed callose synthesis at division sites further suggested a novel pattern of cell plate assembly in gem1. Multiple cell compartments in gem1 adopted vegetative cell fate with regard to lipid body distribution. In the wild type, lipid bodies appear specifically in the vegetative cell, whereas in gem1, lipid bodies accumulated in all cytoplasmic compartments. Our results support the hypothesis that altered pollen cell fate in gem1 results from abnormal inheritance of cell fate determinants as a result of disturbed cytokinesis. PMID- 11402218 TI - Cloning and expression pattern of a gene encoding an alpha-xylosidase active against xyloglucan oligosaccharides from Arabidopsis. AB - An alpha-xylosidase active against xyloglucan oligosaccharides was purified from cabbage (Brassica oleracea var. capitata) leaves. Two peptide sequences were obtained from this protein, the N-terminal and an internal one, and these were used to identify an Arabidopsis gene coding for an alpha-xylosidase that we propose to call AtXYL1. It has been mapped to a region of chromosome I between markers at 100.44 and 107.48 cM. AtXYL1 comprised three exons and encoded a peptide that was 915 amino acids long, with a potential signal peptide of 22 amino acids and eight possible N-glycosylation sites. The protein encoded by AtXYL1 showed the signature regions of family 31 glycosyl hydrolases, which comprises not only alpha-xylosidases, but also alpha-glucosidases. The alpha xylosidase activity is present in apoplastic extractions from Arabidopsis seedlings, as suggested by the deduced signal peptide. The first eight leaves from Arabidopsis plants were harvested to analyze alpha-xylosidase activity and AtXYL1 expression levels. Both increased from older to younger leaves, where xyloglucan turnover is expected to be higher. When this gene was introduced in a suitable expression vector and used to transform Saccharomyces cerevisiae, significantly higher alpha-xylosidase activity was detected in the yeast cells. alpha-Glucosidase activity was also increased in the transformed cells, although to a lesser extent. These results show that AtXYL1 encodes for an apoplastic alpha-xylosidase active against xyloglucan oligosaccharides that probably also has activity against p-nitrophenyl-alpha-D-glucoside. PMID- 11402219 TI - Use of nucleic acid amplification tests in investigating child sexual abuse. PMID- 11402220 TI - Chlamydia trachomatis and cancer. PMID- 11402221 TI - Sector-wide approaches and STI control in Africa. PMID- 11402223 TI - Cohort studies in sexual health. PMID- 11402222 TI - Mitochondrial toxicity and HIV therapy. AB - Nucleoside reverse transcriptase inhibitors (NRTIs) remain the cornerstone of highly active antiretroviral therapy (HAART) combination regimens. However, it has been known for some time that these agents have the potential to cause varied side effects, many of which are thought to be due to their effects on mitochondria. Mitochondria, the key energy generating organelles in the cell, are unique in having their own DNA, a double stranded circular genome of about 16 000 bases. There is a separate enzyme present inside the cell that replicates mitochondrial DNA, polymerase gamma. NRTIs can affect the function of this enzyme and this may lead to depletion of mitochondrial DNA or qualitative changes. The study of inherited mitochondrial diseases has led to further understanding of the consequences of mutations or depletion in mitochondrial DNA. Key among these is the realisation that there may be substantial heteroplasmy among mitochondria within a given cell, and among cells in a particular tissue. The unpredictable nature of mitochondrial segregation during cellular replication makes it difficult to predict the likelihood of dysfunction in a given tissue. In addition, there is a threshold effect for the expression of mitochondrial dysfunction, both at the mitochondrial and cellular level. Various clinical and in vitro studies have suggested that NRTIs are associated with mitochondrial dysfunction in different tissues, although the weight of evidence is limited in many cases. The heterogeneity in the tissues affected by the different drugs raises interesting questions, and possible explanations include differential distribution or activation of these agents. This article reviews the major recognised toxicities associated with NRTI therapy and evidence for mitochondrial dysfunction in these complications. Data were identified through searching of online databases including Medline and Current Contents for relevant articles, along with abstracts and posters from recent conferences in the HIV and mitochondrial fields. PMID- 11402224 TI - An investigation into the pathogenesis of vulvo-vaginal candidosis. AB - OBJECTIVE: To monitor yeasts isolated from women during and between episodes of recurrent vulvo-vaginal candidosis (VVC) to determine whether vaginal relapse or re-infection occurred. METHODS: Women presenting at the genitourinary medicine clinic with signs and symptoms of VVC were recruited to the study (n = 121). A vaginal washing, high vaginal swab (HVS) and rectal swab were taken and the women treated with a single 500 mg clotrimazole pessary. Women were asked to re-attend after 1, 4, and 12 weeks, or when the VVC recurred, when vaginal washings and HVS were repeated. Candida isolates recovered were strain typed using the Ca3 probe and their similarity assessed. Antifungal susceptibility to fluconazole and clotrimazole were determined. RESULTS: Of the women recruited, 47 completed the study, either returning for four visits or suffering a recurrence during the study period. Of the 22 women who experienced recurrence, the same strain was responsible for the initial and recurrent episode in 17 women. For the remaining five women, four had strain replacement and one had a change of species. None of the isolates recovered from the women demonstrated resistance to either clotrimazole or fluconazole. CONCLUSIONS: Our findings support the theory of vaginal relapse and thus may support the use of more prolonged courses of antifungal therapy initially to increase the chances of eradication of the yeast. PMID- 11402225 TI - Increase in sexually transmitted infections among homosexual men in Amsterdam in relation to HAART. AB - OBJECTIVES: We investigated if a rise in rectal gonorrhoea and early syphilis among men who have sex with men (MSM) in Amsterdam coincided with the introduction of highly active antiretroviral therapies (HAART) in July 1996 and determined risk factors for these sexually transmitted infections (STI). METHODS: Subjects were patients of the STI clinic of the municipal health service in Amsterdam. Surveillance data (1994-9) represented consultations (n=11 240) of MSM (n=6103). For analyses we used logistic regression. RESULTS: Comparing the periods before and after the introduction of HAART, the infection rate for rectal gonorrhoea increased from 4% to 5.4% (p=.001) and for syphilis, from 0.5% to 0.8% (p = 0.050). Independent risk factors for rectal gonorrhoea (younger age, western nationality, and concurrent infection with another STI) and for early syphilis (non-western nationality and concurrent infection with rectal gonorrhoea) did not change after HAART became available. For rectal gonorrhoea, however, the infection rate increased only among men who had exclusively homosexual contacts (OR 1.38, p<0.01), compared with bisexual men. For early syphilis, the infection rate increased only among men of western nationality (OR 3.38, p<0.01) compared to men of non-western nationality. CONCLUSIONS: Infection rates of rectal gonorrhoea and early syphilis increased, indicating a change in sexual behaviour, possibly as a result of the introduction of HAART. For now, it is important to find out how sexual behaviour is changing and to keep monitoring trends in STIs (including HIV) among MSM in Amsterdam. PMID- 11402226 TI - Partner notification in HIV-1 infection: a population based evaluation of process and outcomes in Scotland. AB - OBJECTIVES: To evaluate the process and outcomes of HIV partner notification (PN) activity in Scotland. DESIGN: Retrospective population based study. SUBJECTS: 114 adults newly diagnosed with HIV infection (index patients) in Scotland between September 1995 and August 1996. SETTING: Healthcare settings in which all 114 new HIV diagnoses were made: 42 (37%) from genitourinary medicine; 32 (28%) infectious diseases; 18 (16%) general practice; and 22 (19%) from other sites. MAIN OUTCOME MEASURES: Number of partners notified and tested up to 9 months after initial diagnosis. RESULTS: Of 114 index patients (IPs), information on current partners was available for 102 (89%). PN was not appropriate for 47 of the 102 IPs. The remaining 55 IPs identified 63 current partners at risk, of whom 51 were notified: 44 underwent HIV testing, which yielded 11 new HIV positive diagnoses. Information on previous partners was available for only 56 IPs (49%). PN was not appropriate for 30 of the 56 IPs; the remaining 26 IPs identified 46 previous partners at risk, of whom 12 were notified: four were tested, but yielded no new diagnoses. CONCLUSIONS: Notification of current partners was performed well and was an effective strategy for identification of HIV positive individuals at a presymptomatic stage. Notification of previous partners was limited. Partner notification was attempted in a wide range of healthcare settings. Given the clinical effectiveness of antiretroviral therapy, partner notification as a tool towards early diagnosis of HIV disease deserves renewed attention. PMID- 11402227 TI - Prevalence and risk factors of cervicovaginal HIV shedding among HIV-1 and HIV-2 infected women in Dakar, Senegal. AB - OBJECTIVES: To assess the risk determinants and prevalence of cervicovaginal shedding of HIV-1 and HIV-2 among women in Dakar, Senegal. METHODS: We conducted a cross sectional study of 153 HIV seropositive female sex workers (FSW) and another 142 HIV seropositive women attending an infectious diseases unit, based on an interview, physical examination, and laboratory screening for major sexually transmitted infections (STI). Cervicovaginal lavage fluid was tested for HIV-RNA by means of nested PCR. Links between cervicovaginal shedding of HIV-1 and HIV-2 and sociodemographic, clinical, and laboratory variables were identified by using odd ratios and 95% confidence intervals. Logistic regression analysis was used to identify independent links with HIV shedding. RESULTS: The detection rate of HIV-RNA in cervicovaginal lavage fluid was low among FSW, with no difference between HIV-1 (7/90: 8%) and HIV-2 (3/48: 6%). The rate was far higher among the other women (41%, 48/117; 33%, 7/21 for HIV-1 and HIV-2, respectively). In multivariate analysis, high plasma viral load (>40 000 copies/ml) (AOR = 2.4 (1.0-5.6) p = 0.04) and basic vaginal pH (AOR = 2.2 (1.3 3.7) p = 0.002) were independently associated with HIV-1 shedding. For HIV-2 a CD4 count < 200 cells x 10(6)/l was the only factor associated with the shedding of HIV-2 (AOR = 9.0 (0.9-93)). The genital shedding rate was higher with HIV-1 than with HIV-2 (OR = 2.1 (0.9-4.8), but this difference disappeared after adjustment for the CD4+ cell count (AOR = 1.2 (0.5-2.9)). CONCLUSION: Advanced disease stage and immunosuppression are the major risk determinants for shedding of both HIV-1 and HIV-2. Basic vaginal pH is also a risk determinant for HIV-1 shedding. PMID- 11402228 TI - The acceptability of urinary LCR testing for Chlamydia trachomatis among participants in a probability sample survey of sexual attitudes and lifestyles. AB - OBJECTIVES: To examine the factors that influence respondents' willingness to participate in urinary testing for Chlamydia trachomatis in a general population feasibility survey of sexual attitudes and lifestyles. METHODS: 199 sexually experienced, 18-44 year old participants, recruited as part of a larger (n=901) methodological study of sexual attitudes and lifestyles, were invited to provide a urine sample for chlamydial infection testing using ligase chain reaction (LCR) techniques. Analysis of the survey data and in-depth qualitative interviews were undertaken to explore the factors that influenced participants' decisions to participate. RESULTS: 143/199 (72%) participants agreed to provide a urine sample. The likelihood of providing a urine sample was reduced if other individuals were present in the home at the time of interview (OR 0.42, 95% confidence interval 0.20-0.90, p=0.03). Trust and rapport with the interviewer, understanding the aims of the test, sense of obligation, and perceived importance of the test were identified as additional influencing factors in the in-depth interviews. CONCLUSIONS: Survey respondents' uncertainty or embarrassment at participating in urine testing can be overcome if they are well informed, motivated by the potential health gain, and briefed by trained and confident interviewers. PMID- 11402229 TI - Symptomatic HIV seroconverting illness is associated with more rapid neurological impairment. AB - OBJECTIVES: To establish whether symptomatic seroconverting illness in HIV infected people is associated with more rapid development of neurological impairment. METHODS: 166 HIV infected subjects with a known date of HIV infection enrolled in a longitudinal study of neurocognitive function were stratified by whether or not they had experienced a symptomatic serconverting illness. RESULTS: 29 of 166 (17.5%) dated HIV seroconverters had a history of symptomatic seroconverting illness. Though baseline neurocognitive function was similar, subjects with a symptomatic seroconverting illness developed clinical neurocognitive impairment significantly more rapidly than their asymptomatic counterparts in a survival analysis model (636 v 1075 days till impaired). CONCLUSION: Symptomatic seroconverting illness predisposes to more rapid neurocognitive impairment. PMID- 11402230 TI - Deproteinised latex condoms are well tolerated by latex allergic patients. PMID- 11402231 TI - Graded challenge in an aciclovir allergic patient. AB - An immunocompetent woman presented with a hypersensitivity skin reaction following suppressive therapy with aciclovir for recurrent culture proved genital herpes simplex virus infection. She developed a similar reaction when treatment was changed to famciclovir. Without antiviral suppression her recurrences were frequent and distressing. Graded challenge was performed and she became tolerant to aciclovir. She successfully continued suppressive therapy for 1 year with no further hypersensitivity reactions or recurrences. PMID- 11402232 TI - Relation of health literacy to gonorrhoea related care. AB - OBJECTIVE: To assess the relation between health literacy and receipt of a screening test for gonorrhoea in the past year. METHODS: Study design was multisite, cross sectional survey of subjects enrolled from clinics, from community based organisations, and by street intercept. Data were obtained using face to face interview. The dependent variable was self reported receipt of a test for gonorrhoea in the past year. Health literacy was measured by the Rapid Estimate of Adult Literacy in Medicine (REALM), recoded to represent 8th grade or lower reading or 9th grade and higher reading level. Statistical analyses were adjusted to account for selection bias in literacy assessment. RESULTS: 54% of the sample reported at least one gonorrhoea test in the previous year. 65% of the sample read at a 9th grade level or higher. REALM score was moderately correlated with the respondent's years of education. After adjustment for missing REALM data, past suspicion of gonorrhoea, self inspection for gonorrhoea, self efficacy for care seeking, REALM score of 9th grade reading level or higher, and younger age were independently associated with gonorrhoea testing in the previous year. For the average respondent, REALM reading grade level of 9th grade or higher is associated with a 10% increase in the probability of having a gonorrhoea test in the past year. CONCLUSIONS: Low literacy appears to pose a barrier to care for sexually transmitted infections such as gonorrhoea. PMID- 11402233 TI - Combined topical flucytosine and amphotericin B for refractory vaginal Candida glabrata infections. AB - Patients with vaginitis due to highly azole resistant Candida glabrata can be particularly difficult to treat. We describe three cases of longstanding vaginal candidiasis due to C glabrata. These had failed to respond to local and systemic antifungals. Flucytosine (1 g) and amphotericin B (100 mg) formulated in lubricating jelly base in a total 8 g delivered dose, was used per vagina once daily for 14 days with significant improvement, both clinically and microbiologically. PMID- 11402234 TI - The control of syphilis, a contemporary problem: a historical perspective. AB - The control of syphilis in the United Kingdom and United States has been managed in different ways in each country over the course of the last century. Older more established measures including contact tracing, serological surveillance, and health education strategies together with effective antibiotic therapy have had some success. However, changing social structures on both sides of the Atlantic Ocean have brought newer mathematical and epidemiological methods to the fore. This review looks at the past management of syphilis control in the United Kingdom and United States, and speculates on future prospects for disease management in these countries. PMID- 11402235 TI - The antibiotic susceptibility of Neisseria gonorrhoeae isolated in Ulaanbaatar, Mongolia. PMID- 11402236 TI - Prevalence of STDs and HIV infection among immigrant sex workers attending an STD centre in Bologna, Italy. PMID- 11402237 TI - Wood for the trees! PMID- 11402239 TI - Chlamydia trachomatis reinfection rate: a forgotten aspect of female genital chlamydia management. PMID- 11402240 TI - The Society of Apothecaries Diploma examination in Genitourinary Medicine: death of the viva voce? PMID- 11402241 TI - Methicillin resistant Staphylococcus aureus (MRSA) balanoposthitis in an insulin dependent diabetic male. PMID- 11402242 TI - HIV positive and negative homosexual men have adopted different strategies for reducing the risk of HIV transmission. PMID- 11402243 TI - Detection of chlamydia on meatal swabs. PMID- 11402244 TI - Chaperoning male patients. PMID- 11402245 TI - A mobile phone text message and Trichomonas vaginalis. PMID- 11402246 TI - Third trimester screening or safer sex to prevent mother to child transmission of HIV. PMID- 11402247 TI - Lichen sclerosus of the glans is significantly associated with penile carcinoma. PMID- 11402248 TI - Economic advantages of ligase chain reaction for diagnosis of genital Chlamydia trachomatis infection in GUM clinic attenders. PMID- 11402249 TI - Minimal influence of carbohydrate ingestion on the immune response following acute resistance exercise. AB - The effect of carbohydrate supplementation (CHO) on the lymphocyte response to acute resistance exercise was examined in 10 resistance-trained males. Subjects completed a randomized double-blind protocol with sessions separated by 14 days. The exercise session consisted of a high intensity, short rest interval squat workout. Subjects consumed 1.0 g a kg body mass(-1) CHO or an equal volume of placebo (PLC) 10 min prior to and 10 min following exercise. Blood was collected at rest (REST), immediately post exercise (POST), and at 1.5 hours and 4.0 hours of recovery, and analyzed for plasma glucose, serum cortisol, leukocyte subsets, and phytohemagglutinin (PHA)-stimulated lymphocyte proliferation. A significant Treatment 3 Time effect was observed for lymphocyte proliferation between CHO and PLC, but post hoc analyses revealed no between-treatment differences at any post exercise time point. Lymphocyte proliferation was significantly depressed below REST at POST (-39.2% for PLC, -25.7% for CHO). Significant fluctuations in leukocyte subset trafficking were observed for both treatments at POST, 1.5 hours, and 4.0 hours. Plasma glucose was significantly increased POST in CHO compared to PLC. Cortisol was significantly increased from REST to POST in both treatments. These data support a minimal effect of carbohydrate ingestion on the lymphocyte response to high-intensity resistance exercise. PMID- 11402250 TI - Body composition profiles of elite American heptathletes. AB - This study characterized body composition profiles of elite American heptathletes and cross-validated skinfold (SKF) and bioelectrical impedance analysis (BIA) field method equations for estimation of percent body fat (%Fat) using dual energy x-ray absorptiometry (DXA) as the criterion. Weight, height, fat mass (FM), fat-free mass (FFM), bone mineral density (BMD), and %Fat were measured in 19 heptathletes using standard measurement protocols for DXA, SKFs and BIA. The ages, heights, and weights were respectively 25.5 +/- 3.5 years, 175.0 +/- 6.6 cm, 67.3 +/- 7.1 kg. DXA estimates of mean +/- SD values for body composition variables were 57.2 +/- 6.1 kg FFM, 10.1 +/- 2.6 kg FM, 114 +/- 7% BMD for age/racial reference group, and 15 +/- 3.0 %Fat. Ranges of bias values for %Fat (DXA minus SKF or BIA) were, respectively, -0.5 to 1.6% and -5.5 to -1.2%. Ranges for standard errors of estimate and total errors were, respectively, SKF 2.4 2.5%, 2.4 - 2.8% and BIA 3.0%, 5.0-6.5%. Regression analyses of the field methods on DXA were significant (p < .05) for all SKF equations but not BIA equations. This study demonstrates that elite American heptathletes are lean, have high levels of BMD, and that SKF equations provide more accurate estimates of %Fat relative to DXA than estimates from BIA equations. PMID- 11402251 TI - Nutrition knowledge, opinions, and practices of coaches and athletic trainers at a division 1 university. AB - The purpose of this investigation was to assess nutrition knowledge, opinions, and practices of coaches and trainers at a Division I university. Participants (n = 53) completed questionnaires regarding nutrition knowledge, opinions, and practices. Descriptive statistics and analysis of variance were used to analyze data. Overall, participants responded correctly to 67% of nutrition knowledge questions. Participants who coached/trained female athletes tended to score better than respondents who coached/trained male athletes. Strength and conditioning coaches and participants with greater than 15 years of experience scored higher than other participants. Nutrition opinions/practices responses revealed that nutritional supplements were provided for all but 6% of participants' athletes. Participants rated body weight as more important than body composition to athletes' performances. Over 30% of participants perceived at least one case of disordered eating within the past year. Some participants (53%) felt that athletes may consume more nutritious meals on team-sponsored trips if given larger food allowances. Thirty percent of participants reported dietitians were available to them; the same percentage reported utilizing dietitians. Coaches and trainers are knowledgeable about some appropriate nutritional recommendations, but registered dietitians or qualified sports nutrition professionals may complement the nutrition-related education and counseling of athletes (23). PMID- 11402252 TI - Interactions among dietary fat, mineral status, and performance of endurance athletes: a case study. AB - In a pilot study, performance measures and mineral metabolism were assessed in 3 male endurance cyclists who consumed isoenergetic, isonitrogenous diets for 28 day periods in a randomized, crossover design in which dietary carbohydrate, polyunsaturated, or saturated fat contributed about 50% of daily energy intake. Peak aerobic capacity [62 ml/(kg a min)] was unaffected by diet. Endurance capacity at 70-75% peak aerobic capacity decreased with the polyunsaturated fat diet. Copper retention tended to be positive only with saturated fat. Less iron and zinc were retained (intake - losses), and fecal losses of these minerals increased with the polyunsaturated fat. Blood biochemical measures of trace element nutritional status were unaffected by diet, except serum ferritin, which tended to decrease during consumption of the polyunsaturated fat diet. These preliminary results suggest that diets high in polyunsaturated fat, particularly linoleic acid, impair absorption and utilization of iron and zinc, and possibly magnesium, and may reduce endurance performance. PMID- 11402253 TI - Food sources of calcium in a sample of African-American an Euro-American collegiate athletes. AB - Adequate calcium intake is integral to bone health as well as for optimal athletic performance. This study was conducted to investigate: (a) food sources of calcium in a sample of collegiate athletes, (b) gender and/or ethnic differences in food sources of calcium, and (c) whether athletes that derive less of their calcium intake from dairy sources increase their calcium intake from supplements or other food sources. Participants were African-American and Euro American NCAA Division 1-A athletes. Eighty-five men and 59 women participated. Calcium intake for the previous 7-day period was assessed with a brief calcium screen. Men consumed significantly more calcium than women (1,354 vs. 898 mg/day), with female cross-country runners exhibiting the lowest average intake (605 mg/day). Both men and women obtained the majority of their calcium from dairy products and mixed dishes, while men consumed significantly more calcium fortified foods. Several gender and ethnic interactions for calcium intake from food groups were found. Mean total dairy calcium intake was found to vary according to total calcium intake in men, and supplemental calcium was not used to augment low dairy intakes of calcium in any group. While African-Americans and Euro-Americans athletes were consuming similar levels of calcium, the female athletes in the sample did not get adequate amounts. PMID- 11402254 TI - High-fat diet versus habitual diet prior to carbohydrate loading: effects of exercise metabolism and cycling performance. AB - We examined the effects of a high-fat diet (HFD-CHO) versus a habitual diet, prior to carbohydrate (CHO)-loading on fuel metabolism and cycling time-trial (TT) performance. Five endurance-trained cyclists participated in two 14-day randomized cross-over trials during which subjects consumed either a HFD (> 65% MJ from fat) or their habitual diet (CTL) (30 +/- 5% MJ from fat) for 10 day, before ingesting a high-CHO diet (CHO-loading, CHO > 70% MJ) for 3 days. Trials consisted of a 150-min cycle at 70% of peak oxygen uptake (VaO2peak), followed immediately by a 20-km TT. One hour before each trial, cyclists ingested 400 ml of a 3.44% medium-chain triacylglycerol (MCT) solution, and during the trial, ingested 600 ml/hour of a 10% 14C-glucose + 3.44% MCT solution. The dietary treatments did not alter the subjects' weight, body fat, or lipid profile. There were also no changes in circulating glucose, lactate, free fatty acid (FFA), and b-hydroxybutyrate concentrations during exercise. However, mean serum glycerol concentrations were significantly higher (p < .01) in the HFD-CHO trial. The HFD CHO diet increased total fat oxidation and reduced total CHO oxidation but did not alter plasma glucose oxidation during exercise. By contrast, the estimated rates of muscle glycogen and lactate oxidation were lower after the HFD-CHO diet. The HFD-CHO treatment was also associated with improved TT times (29.5 +/- 2.9 min vs. 30.9 +/- 3.4 min for HFD-CHO and CTL-CHO, p <.05). High-fat feeding for 10 days prior to CHO-loading was associated with an increased reliance on fat, a decreased reliance on muscle glycogen, and improved time trial performance after prolonged exercise. PMID- 11402255 TI - Free living energy expenditure in post menopausal women before and after exercise training. AB - The aim of the study was to determine the effects of 8 weeks of moderate exercise training, on 24-hour free living energy expenditure in previously sedentary post menopausal women. The experimental group (EX) included 9 women. Ten non exercising control subjects (CON) were recruited to undergo pre- and post testing. Estimated total daily energy expenditure (TDEE), total 24-hour heart beats (HB), total energy intake (TEI), resting metabolic rate, maximal oxygen consumption (VaO2max), body composition, and submaximal heart rate were measured before and after the exercise intervention. Body composition did not change (body fat % in CON 34.0 +/- 4.0% vs. 33.9 +/- 3.6% and EX 34.1 +/- 4.0% vs. 34.0 +/- 3.4%). Mean submaximal heart rate during steady-state exercise in EX was lower after training compared to CON (p < .05); however, VaO2max did not increase significantly (CON 1.96 +/- 0.23 vs. 1.99 +/- 0.24 l L O2/min and EX 1.86 +/- 0.39 vs. 1.94 +/- 0.30 L O2/min). Neither estimated TDEE (CON, 11.6 +/- 2.0 vs. 11.4 +/- 2.78 MJ; and EX 11.4 +/- 3.3 vs. 11.5 +/- 2.5 MJ, pre vs. post, respectively), RMR (CON 134.2 +/- 9.4 vs. 136.9 +/- 15.0 KJ/kgFFM/day, and EX 138.4 +/- 6.4 vs. 140.7 +/- 14.2 KJ/kgFFM/day, pre vs. post, respectively), TEI (CON 7.9 +/- 2.2 vs. 8.2 +/- 2.5 MJ, and EX 9.4 +/-1.6 vs. 8.3 +/- 2.8 MJ), nor HB (CON 110,808 +/- 12,574 vs. 107,366 +/- 12,864 beats, and EX 110,188 +/- 9,219 vs. 114,590 +/- 12,750 beats) change over 8 weeks in either group. These data suggest that a moderate exercise program may not impact on TDEE, RMR, TEI, or HB in previously sedentary, older women. PMID- 11402256 TI - The effect of low-carbohydrate diet on the pattern of hormonal changes during incremental, graded exercise in young men. AB - The purpose of this study was to discover whether severe dietary carbohydrate (CHO) restriction modifies the relationship between exercise intensity and hormonal responses to exercise. Changes in the plasma adrenaline (A), noradrenaline (NA), growth hormone (hGH), testosterone (T), and blood lactate (LA) during an incremental exercise performed until volitional exhaustion were determined in 8 physically active volunteers after 3 days on low CHO (< 5% of energy content; L-CHO) and isocaloric mixed (M) diets. Following L-CHO diet, the basal plasma A, NA, and hGH concentrations were increased, whilst T and LA levels were decreased. During exercise all the hormones increased exponentially, with thresholds close to that of LA. Neither the magnitude nor the pattern of the hormonal changes were affected by L-CHO diet except the NA threshold, which was lowered. Blood LA response to exercise was diminished and LA threshold was shifted towards higher loads by L-CHO diet. It is concluded that restriction of CHO intake (a) does not affect the pattern of changes in plasma A, hGH, and T concentrations during graded exercise but lowers NA threshold, indicating increased sensitivity of the sympathetic nervous system to exercise stimulus; (b) alters the basal and exercise levels of circulating hormones, which may have an impact on the balance between anabolic and catabolic processes and subsequently influence the effectiveness of training. PMID- 11402257 TI - Nutritional supplements as a source for positive doping cases? AB - We report the findings of the analysis of 75 different nutritional supplements bought through the internet. Seven products (all from the class of prohormones) contained other hormone substances than indicated on the labels, and two further products contained ephedrine and caffeine without a clear indication on the labels. PMID- 11402258 TI - Gleevec (STI-571) for chronic myeloid leukemia. PMID- 11402259 TI - Ziprasidone (Geodon) for schizophrenia. PMID- 11402261 TI - Use of telehealth to deliver home health services. PMID- 11402262 TI - Home health care, ethics, and the family. PMID- 11402263 TI - Positive inotropic drug infusions for the patient with heart failure. PMID- 11402264 TI - Outliers in home care. PMID- 11402265 TI - HCFA reform. Health Care Financing Administration. PMID- 11402266 TI - The road to success in PPS: guiding care for efficiency and effectiveness. Prospective payment system. AB - This article features one organization's attempt to interpret and form operational definitions for the initiation of the prospective payment system (PPS) by October 1, 2000. Data collection, initiated with the October 28, 1999, proposed regulations, showed inconsistent use of resources when patient needs were similar based on the initial 20 indicators used to calculate acuity and payment levels in PPS. A decision to use a clinical guidance system with the potential to control outcomes and use rates resulted in the development of a system called Care Guides. These tools help home care staff develop and implement efficient and effective home care plans. PMID- 11402267 TI - Palliative care: long-term solution for long-term care. Part I. PMID- 11402268 TI - Nonsteroidal anti-inflammatory drugs: the need for assessment and education. PMID- 11402269 TI - Evidence-based endodontics: the one-visit treatment idea. PMID- 11402270 TI - Image resolution. PMID- 11402272 TI - Human papillomavirus as a risk factor for oral squamous cell carcinoma: a meta analysis, 1982-1997. AB - OBJECTIVE: Human papillomavirus (HPV) infection is a significant risk factor for uterine cervical carcinoma. However, the role of HPV infection in oral squamous cell carcinoma (OSCC) is less well defined. To determine the significance of the relationship of this virus in the progressive development of oral cancer, we estimated the risk of HPV detection in normal oral mucosa, precancerous oral tissue, and oral carcinoma using meta-analysis. STUDY DESIGN: Case reports and clinical series published in English-language journals were retrieved by searching MEDLINE (January 1980-August 1998). Review articles were also examined to identify additional studies. Studies that used biochemical, immunologic, microscopic, or molecular analyses to detect HPV in tissue or cells derived from normal oral mucosa (n = 25), benign leukoplakia (n = 21), intraepithelial neoplasia (ie, dysplasia and carcinoma in situ; n = 27), and oral cancer (n = 94) were included in the meta-analysis. Information on sample size, age, sex, method of tissue preservation (ie, fresh, frozen, paraffin-embedded), assay, primer amplification region (early, late), high-risk versus low-risk genotype, and use of tobacco or alcohol was abstracted by one author (C.S.M.). RESULTS: Data from 94 reports that analyzed 4680 samples were included in the meta-analysis. Analyses made by means of a random-effects model with and without adjustments for assay sensitivity showed increased probability of HPV detection in tissue with precancerous and cancerous features compared with normal mucosa. The likelihood of detecting HPV in normal oral mucosa (10.0%; 95% confidence interval [CI], 6.1% 14.6%) was significantly less than of detecting benign leukoplakia (22.2%; 95% CI, 15.7%-29.9%), intraepithelial neoplasia (26.2%; 95% CI, 19.6%-33.6%), verrucous carcinoma (29.5%; 95% CI, 23%-36.8%), and OSCC (46.5%; 95% CI, 37.6% 55.5%). Adjustment of findings for differences in assay sensitivity indicated that these estimates may be conservative. Overall, HPV was between 2 and 3 times more likely to be detected in precancerous oral mucosa and 4.7 times more likely to be detected in oral carcinoma than in normal mucosa. The pooled odds ratio for the subset of studies directly comparing the prevalence of HPV in normal mucosa and OSCC was 5.37, confirming the trend observed in the overall sample. The probability of detecting high-risk HPVs in OSCCs was 2.8 times greater than that of low-risk HPVs. CONCLUSION: This meta-analysis indicates that HPV is detected with increased frequency in oral dysplastic and carcinomatous epithelium in comparison with normal oral mucosa. The findings provide further quantitative evidence that oral infection with HPV, particularly with high-risk genotypes, is a significant independent risk factor for OSCC. PMID- 11402273 TI - Treatment of a central giant cell lesion of the mandible with intralesional glucocorticosteroids. PMID- 11402274 TI - Accuracy of the Third Molar Eruption Predictor in predicting eruption. AB - OBJECTIVE: To evaluate the possibility of applying the Third Molar Eruption Predictor to all panoramic radiographs. STUDY DESIGN: Panoramic radiographs were retrospectively analyzed from a 4-year follow-up study of third molars carried out at the University of Copenhagen, Denmark. The radiographs, taken at a mean age of 20.6 years, included 45 unerupted or partially erupted mandibular third molars in 28 subjects. Because the device was calibrated both with simple proportions and by use of the methods of Bayes' Decision Theory, the separation point of the device was therefore adjusted at 12 mm from the distal surface of the second molar. RESULTS: The predictions of future eruption or impaction made with the calibrated device and the actual clinical outcome 4 years later were in conformity for 80% of the mandibular third molars. CONCLUSION: The Third Molar Eruption Predictor may be applied to all panoramic radiographs, but it seems to require calibration before use. PMID- 11402275 TI - Evaluation of oral antimicrobial agent levels in tooth extraction sites. AB - OBJECTIVE: The purpose of this study was to evaluate various oral antimicrobial agent levels in tooth extraction sites. STUDY DESIGN: The concentration of dental alveolar blood in extraction wounds after the oral administration of talampicillin (500 mg), cefaclor (500 mg), cefteram pivoxil (200 mg), cefuroxime axetil (250 mg), cefdinir (200 mg), and ofloxacin (100 mg) was determined in 338 patients and was assessed on the basis of its antimicrobial activity against Streptococcus isolated in odontogenic infections. RESULTS: The percentage of patients whose concentrations exceeded the minimum inhibitory concentration for 90% of Streptococcus was 62.5% to 100% for talampicillin at 30 to 360 minutes, 0% to 12.5% for cefaclor at 30 to 360 minutes, 18.2% to 100% for cefteram pivoxil at 30 to 480 minutes, 50% to 100% for cefuroxime axetil at 30 to 480 minutes, 0% to 50% for cefdinir at 16 to 290 minutes, and 0% to 40% for ofloxacin at 30 to 480 minutes. CONCLUSION: These results indicate that talampicillin, cefteram pivoxil, and cefuroxime axetil have minimum inhibitory concentration levels for 90% of Streptococcus in tooth sockets. PMID- 11402276 TI - Ameloblastoma: a clinical, radiographic, and histopathologic analysis of 71 cases. AB - OBJECTIVES: The purpose of this study was to compare the clinical, radiologic, and histopathologic features of 71 intraosseous ameloblastomas. STUDY DESIGN: Data with respect to the patients' ages, sex, tumor locations, and surgical treatment history, as well as the radiographic findings and number of recurrences, were analyzed. The histologic types of and radiologic findings regarding tumors with higher recurrence rates were also investigated. RESULTS: The patients' ages at biopsy ranged from 11 to 70 years (mean, 30.4 years). Thirty-nine (54.9%) of the 71 subjects were males, and 32 (45.1%) were females. Sixty-two (87.3%) of the 71 ameloblastomas were located in the mandible. Swelling was the most common symptom and was experienced by 27 (38.0%) patients. Radiographically, 42 (59.2%) of the 71 tumors were unilocular with a well demarcated border. Of the remaining 29 cases, 14 were multilocular, 2 were of soap-bubble shape, and 13 were unknown in appearance. The most common histologic pattern was plexiform, rather than follicular or acanthomatous. Sixteen cases of ameloblastoma had developed in a cyst. The overall recurrence rate was 21.1%, and the average age of the patient at recurrence was 26.4 years. CONCLUSIONS: When the diagnosis of ameloblastoma in young people remains in doubt after clinical and radiologic examination, a biopsy is necessary. Long-term follow-up at regular intervals after surgery is also recommended. PMID- 11402277 TI - Anxiety and pain suppress the natural killer cell activity in oral surgery outpatients. AB - OBJECTIVE: Psychological stress has an influence on natural killer cell (NK) activity, which plays a central role in protection against microbial infection and cancer. Anxiety concerning cancer is a typical type of psychological stress observed in patients and is associated with various diseases. In this study, we examined whether anxiety about cancer reduces the NK activity or quality of life (QOL), or both, of outpatients. PATIENTS AND METHODS: NK activity was determined by means of a 4-hour 51 chromium-release assay, and a QOL questionnaire by the World Health Organization called QOL-BRIEF was also used. One hundred forty-four patients were asked a variety of questions about anxiety with respect to cancer and pain on their first visit to the outpatient clinic of our Department of Oral Surgery. RESULTS: The NK activity in patients with anxiety about cancer was significantly lower (P < .001) than that in those with no such anxiety. In contrast, NK activity was not influenced by any other factor tested, including differences in diseases and QOL scores. CONCLUSION: Anxiety about cancer may be one of the primary factors for suppression of NK activity in oral surgery outpatients. PMID- 11402278 TI - Candida dubliniensis in radiation-induced oropharyngeal candidiasis. AB - Candida dubliniensis is a recently described species that has been shown to cause oropharyngeal candidiasis in patients with HIV. We present a detailed evaluation of a patient undergoing head and neck radiation for oral cancer who developed oropharyngeal candidiasis from a mixed infection of C dubliniensis and Candida albicans. To our knowledge, this is the first described case of C dubliniensis contributing to oropharyngeal candidiasis in this patient population. PMID- 11402279 TI - In vitro studies of the efficacy of antimicrobials against fungi. AB - OBJECTIVE: The purpose of this study was to compare the efficacy of Listerine Antiseptic, Tartar Control Listerine Antiseptic, and Peridex mouthrinses and a 0.2% chlorhexidine digluconate solution against known pathogenic fungi. STUDY DESIGN: Standardized methods were used to compare the antimicrobial efficacy of the above agents versus representative fungal species. Minimum inhibitory concentration-minimum fungicidal concentrations in macrobroth dilutions, suspension kill-time, and effectiveness against an artificial biofilm-attached population were studied. RESULTS: All antimicrobials tested were effective against the fungal species under investigation at the concentration available commercially. Listerine Antiseptic showed a greater efficacy against attached artificial biofilm populations than the other antimicrobials tested. CONCLUSIONS: Listerine Antiseptic, Tartar Control Listerine Antiseptic, and Peridex mouthrinses show promise as a means to control the pathogenic fungal species under investigation and may have applications to reduce oral colonization. PMID- 11402280 TI - Randomized clinical trial of primary treatment for temporomandibular joint disk displacement without reduction and without osseous changes: a combination of NSAIDs and mouth-opening exercise versus no treatment. AB - OBJECTIVE: The effectiveness of nonsteroidal anti-inflammatory drugs (NSAIDs) and physical therapy for disk displacement without reduction is unknown. This study compared this treatment method with nontreatment controls. METHODS: Sixty patients with painful disk displacement without reduction and without osseous changes were randomly divided in 2 groups, consisting of NSAID and physical therapy and a nontreated control group. Both groups were observed at 2 weeks and, for those patients who did not show any improvement, again at 4 weeks. RESULTS: There was 60% improvement in the treatment group compared with 33% in the control group during the entire 4 weeks of the study. The number needed to treat for benefit was 3.75, with a 95% CI 2.103 to 65.935. CONCLUSIONS: A combination of NSAID and physical therapy for 4 weeks is effective as a primary treatment of patients with disk displacement without reduction and without osseous changes. PMID- 11402281 TI - Cardiac risk stratification for postmyocardial infarction dental patients. AB - Traditional dental management guidelines of myocardial infarction survivors mandate a 6-month waiting period before elective treatment can be considered. Technological advances in cardiac disease diagnosis, management, and revascularization treatment may make this older mandatory 6-month waiting period obsolete. The purposes of this literature review are to provide an overview of the historical development of cardiac risk stratification and discuss current developments and guidelines in cardiac risk assessment. We hope that this review and update will stimulate the development of updated dental guidelines for treating the cardiac patient. PMID- 11402282 TI - Adolescent gingival squamous cell carcinoma: Report of a case and review of the literature. AB - In this article we describe a case of adolescent squamous cell carcinoma of the gingiva. The tumor presented as an ulcerative lesion of the gingiva that was originally thought to be pseudoepitheliomatous hyperplasia. The pathologic differentiation between squamous cell carcinoma and pseudoepitheliomatous hyperplasia is discussed and a review of the literature with respect to pediatric and adolescent oral squamous cell carcinoma is presented. The reported case illustrates the difficulties that may be encountered in attempting to histopathologically distinguish between pseudoepitheliomatous hyperplasia and well-differentiated squamous cell carcinoma of the oral cavity. PMID- 11402283 TI - Heterotopic gastrointestinal cyst partially lined with dermoid cyst epithelium. AB - We report a rare heterotopic gastrointestinal cyst located in the right submandibular/submental area with histopathologic features that included portions resembling a dermoid cyst. Some theories of pathogenesis are discussed, and an origin of this lesion in entrapped undifferentiated endodermal cells is suggested. PMID- 11402284 TI - Intraoral salivary duct carcinoma: case report with immunohistochemical observations. AB - Salivary duct carcinoma is an uncommon malignant salivary gland tumor that occurs predominantly in the parotid gland. Oral involvement is extremely rare, with few cases having been reported in the literature. The tumor is characterized by an aggressive behavior and has a poor prognosis. We describe a case of salivary duct carcinoma arising in the hard palate of a 63-year-old man. Immunohistochemical analysis revealed that tumor cells tested positive for cytokeratin, epithelial membrane antigen, proliferating cell nuclear antigen, Ki67, p53, laminin, and collagen IV. Despite radical surgical resection, bilateral neck dissection, and postoperative radiotherapy, liver metastases developed, and the patient subsequently died of his disease. PMID- 11402285 TI - Metastatic calcification of the nasal septum presenting as an intraoral mass: a case report with a review of the literature. AB - Metastatic calcification is a pathologic condition characterized by deposition of calcified product in otherwise normal tissues as a result of hyperphosphatemia with or without concurrent hypercalcemia. Metastatic calcification presenting clinically as an oral lesion is extremely rare. To date, only 7 cases of metastatic calcification involving the oral soft tissues have been described. This report describes a case of metastatic calcification of the nasal septum presenting as a mass of the anterior maxillary vestibule in a patient with end stage renal disease. The case reported is only the second example with nasal septum involvement. A brief review of the clinical and histopathologic features of previously reported intraoral cases is also presented. PMID- 11402286 TI - Comparison of clinical outcome of periapical surgery in endodontic and oral surgery units of a teaching dental hospital: a retrospective study. AB - OBJECTIVES: The aims of this retrospective study were (1) to compare the outcome of periapical surgery performed in endodontic and in oral surgery units of a teaching dental hospital and (2) to evaluate the influence of factors affecting outcome. STUDY DESIGN: A total of 176 teeth (endodontic unit, 83; oral surgery unit, 93) surgically treated more than 4 years previously were examined clinically and radiographically by means of strict criteria. Multiple logistic regression analysis was used. RESULTS: The rate of complete healing for patients treated in the endodontic unit (37.4%) was significantly (P = .009) higher than that for patients treated in the oral surgery unit (19.4%). The technical quality of surgery (P < .001), placement of root-end filling (P = .039), absence of a preoperative periapical lesion (P = .042), absence of a post (P = .047), and presence of an adequate coronal restoration (P = .056, odds ratio = 3.71) had significant effects on treatment outcome. CONCLUSION: The technical quality of periapical surgery, the presence of a periapical lesion, and adequate apical and coronal seal are important prognostic determinants of successful periapical surgery. PMID- 11402287 TI - Bone repair of experimentally induced through-and-through defects by Gore-Tex, Guidor, and Vicryl in ferrets: a pilot study. AB - OBJECTIVE: The purpose of this pilot study was to evaluate whether improved bone regeneration can be achieved in experimentally induced through-and-through mandibular defects in ferrets and whether there is a quantitative and qualitative difference of regenerated bone with different guided tissue regeneration membranes. STUDY DESIGN: Through-and-through osseous defects were created at the apices of 16 mandibular premolars of 8 ferrets. The transosseous defects were covered with Gore-Tex, Vicryl, or Guidor membranes. As control, the defects were covered with mucoperiosteal flaps without any membranes. RESULTS: The control group showed ingrowth of sulcular epithelium into the defect. The Gore-Tex group showed good bone formation, whereas the Vicryl group showed the highest degree of bone formation. Six weeks after the operation, the defect had nearly completely filled with fibered and lamellar bone. Twelve weeks after the operation, mainly lamellar bone was observed. In contrast, the Guidor group was found to have limited bone regeneration. CONCLUSION: The results of this in vivo study suggest that guided tissue regeneration membranes generally promote and improve bone regeneration in osseous defects of endodontic origin. PMID- 11402288 TI - A comparison of Greater Taper files, ProFiles, and stainless steel files to shape curved root canals. AB - OBJECTIVE: The purpose of this study was to evaluate the ability of engine-driven nickel-titanium files to maintain the original curvature of root canals during canal preparation. STUDY DESIGN: A total of 36 simulated curved canals on resin blocks were instrumented with Greater Taper (GT) files, ProFiles, and stainless steel files. With the mounting device, which can reproduce the same position of the resin blocks and the camera, the unprepared and prepared canals were accurately compared by means of a double-exposure photographic technique. The analysis of variance test was used for the statistical analysis of data obtained. RESULTS: The canals prepared by means of a ProFile with a 6% taper up to the working length were excellent in taper and in maintaining the original curvature. The canals prepared with GT files were also found to be excellent in taper and in maintaining the original curvature; however, these canals showed a slight enlargement toward the inner side at the beginning of the curvature. The canals prepared with stainless steel files showed nontapered widening and severe transportation. CONCLUSION: The canals prepared with GT files and ProFiles were excellently tapered and maintained the original curvature of the canals in comparison with the ones prepared with stainless steel files. PMID- 11402289 TI - Possibility of sialographic sonography: a Doppler phantom study. AB - OBJECTIVES: The purpose of this study was to develop a new diagnostic method that has the merits of both sialography and sonography. STUDY DESIGN: Saline solution and various contrast media (Urografin 76%; 100%, 90%, and 67% Lipiodol Ultra Fluide; 5% and 1% barium sulfate; and Levovist) were injected into thin tubes at a rate of approximately 0.001 to 0.1 mL/s. The relationship between the Doppler signal intensity and the kind, concentration, and velocity of the fluid was analyzed. RESULTS: Levovist, 90% and 67% Lipiodol Ultra-Fluide, and the barium sulfate solutions produced Doppler signals. The mixture of Lipiodol Ultra-Fluide and saline solution produced high signals at any concentration, in contrast with the barium sulfate solutions. Signals could be observed at any speed, from the speed of normal sialography down to 0.001 mL/s, and there was a proportional relationship between signal intensity and velocity for all fluids producing signals. CONCLUSION: The fact that we could obtain high signals with several fluids indicates potential clinical diagnostic usefulness of sialographic sonography. PMID- 11402290 TI - Ewing's sarcoma of the mandible: radiologic features with emphasis on magnetic resonance appearance. AB - Ewing's sarcoma is an uncommon malignancy that usually occurs in children. A case of Ewing's sarcoma of the mandible is presented, and the radiologic appearance is described, with special consideration given to the magnetic resonance imaging features. PMID- 11402291 TI - Comparative dose measurements by spiral tomography for preimplant diagnosis: the Scanora machine versus the Cranex Tome radiography unit. AB - Objective. The purpose of this study was to determine the dose profile of the Cranex Tome radiography unit and compare it with that of the Scanora machine. Study design. The radiation dose delivered by the Cranex Tome radiography unit during the cross-sectional mode was determined. Single tooth gaps in regions 3 (16) and 30 (46) were simulated. Dosimetry was carried out with 2 phantoms, a head and neck phantom and a full-body phantom loaded with 142 thermoluminescent dosimeters (TLD) and 280 TLD, respectively; all locations corresponded to radiosensitive organs or tissues. The recorded local mean organ doses were compared with those measured in another study evaluating the Scanora machine.Results. Generally, dose values from the Cranex Tome radiography unit reached only 50% to 60% of the values measured for the Scanora machine. The effective dose was calculated as 0.061 mSv and 0.04 mSv for tooth regions 3 (16) and 30 (46), respectively. Corresponding values for the Scanora machine were 0.117 mSv and 0.084 mSv.Conclusion. Cross-sectional imaging in the molar region of the upper and the lower jaw can be performed with the Cranex Tome unit, which delivers only approximately half of the dose that the Scanora machine delivers. PMID- 11402292 TI - A radiologic analysis of dentigerous cysts and odontogenic keratocysts associated with a mandibular third molar. AB - OBJECTIVE: The purpose of this study was to discriminate radiographically between dentigerous cysts (DCs) and odontogenic keratocysts (OKCs) associated with a mandibular third molar. STUDY DESIGN: The material consisted of panoramic radiographs of dentigerous cysts (44 patients, 45 cysts) and odontogenic keratocysts (15 patients, 16 cysts), all of which were related to a mandibular third molar. The radiographic images were analyzed with reference to the patients' ages and symptoms. RESULTS: The mean age of patients in the OKC group was less than that of patients in the DC group. The mean area of the cysts in the OKC group was larger than that of those in the DC group. The mean distance from the second to the third molar in the DC group was greater than that in the OKC group. Although there was a significant correlation between the area and distance in the DC and OKC groups, the patients' ages did not significantly correlate to the area and distance of either cyst. CONCLUSIONS: The OKCs had a tendency toward rapid growth in the patient's youth but short movement of a third molar compared with the DCs. The DCs and OKCs do not appear to develop gradually from the period when follicles or dental lamina were formed but arise at various periods randomly. PMID- 11402295 TI - Protein transduction: an alternative to genetic intervention? AB - Protein transduction, an emerging technology with potential applications in gene therapy, can best be described as the internalisation of proteins into the cell, from the external environment. This process relies on the inherent property of a small number of proteins and peptides of being able to penetrate the cell membrane. The transducing property of these molecules can be conferred upon proteins which are expressed as fusions with them and thus offers an alternative to gene therapy for the delivery of therapeutic proteins into target cells. This review describes the three most commonly used protein transduction vehicles; the antennapedia peptide, the herpes simplex virus VP22 protein and HIV TAT protein transduction domain. The future prospects for the application of this technology in gene therapy are also discussed. PMID- 11402296 TI - Novel gene delivery systems: complexes of fusigenic polymer-modified liposomes and lipoplexes. AB - We have previously developed the succinylated poly(glycidol)-modified liposome which becomes fusigenic under weakly acidic condition. In this report, we describe that complexation of this pH-sensitive, fusigenic liposome with a lipoplex consisting of 3beta-(N-(N',N'-dimethylaminoethane) carbamoyl)cholesterol, dioleoylphosphatidylethanolamine and plasmid DNA gives efficient gene delivery systems. In this study, we prepared the complexes, which are termed SucPG-complexes, with a positively or negatively charged surface by mixing the lipoplex with varying amounts of the SucPG-modified liposomes. The positively charged SucPG-complexes either bearing or not bearing a cell-specific ligand, transferrin, could transfect HeLa cells efficiently. In contrast, the negatively charged complexes hardly transfected the cells when transferrin was not conjugated to them. However, the negatively charged SucPG complexes bearing transferrin exhibited high transfection ability against HeLa and K562 cells, indicating that this gene delivery was achieved through their binding to the cellular receptors. These transferrin-attached, negatively charged complexes retained the high transfection ability in the presence of serum. Thus, this negatively charged complex may be useful as nonviral vectors in vivo. PMID- 11402297 TI - Central nervous system gene therapy with interleukin-4 inhibits progression of ongoing relapsing-remitting autoimmune encephalomyelitis in Biozzi AB/H mice. AB - Multiple sclerosis (MS) is an immune-mediated inflammatory disease of the central nervous system (CNS) that might benefit from anti-inflammatory therapies. However, systemic delivery of anti-inflammatory drugs in MS patients has so far been disappointing, mostly due to the limited capacity of these molecules to enter the CNS. We injected into the cisterna magna (i.c.) of Biozzi AB/H mice affected by a relapsing-remitting form of experimental autoimmune encephalomyelitis (EAE), the animal model of MS, a non-replicative herpes simplex virus (HSV) type-1-derived vector containing the interleukin (IL)-4 gene (d120:LacZ:IL-4). CNS delivery of the d120:LacZ:IL-4 vector, after EAE onset, induced the in situ production of IL-4 by CNS-resident cells facing the cerebrospinal fluid (CSF) spaces and reduced by 47% (P < 0.02) the disease related deaths. Compared with mice treated with the control d120:lacZ vector, IL 4-treated mice also showed a shorter duration of the first EAE attack, a longer inter-relapse period, and a reduction in the severity and duration of the first relapse. Protection from EAE progression in IL-4-treated mice was associated with activation of microglia in spinal cord areas where mRNA content of the pro inflammatory chemokines, macrophage chemoattractant protein-1 (MCP-1) and Rantes, was reduced and that of the anti-inflammatory cytokine IL-4 was increased. Finally, CNS-infiltrating mononuclear cells from IL-4-treated mice produced lower levels of MCP-1 mRNA compared with control mice. Our results, showing that IL-4 gene delivery using HSV-1 vectors induces protection from EAE by in situ modulating the cytokine/chemokine-mediated circuits sustaining effector cell functions, indicate that the intrathecal 'therapeutic' use of nonreplicative HSV 1-derived vectors containing anti-inflammatory molecules might represent an alternative strategy in inflammatory diseases of the CNS. PMID- 11402298 TI - Efficient repetitive gene delivery to skeletal muscle using recombinant adenovirus vector containing the Coxsackievirus and adenovirus receptor cDNA. AB - To improve adenovirus-mediated gene delivery to skeletal muscle, we have used a recombinant adenovirus vector encoding the human Coxsackievirus and adenovirus receptor (hCAR). Because CAR is expressed at a lower level in rodent myoblasts and muscle fibers than in other tissues, we expected that elevated expression of CAR in skeletal muscle would improve the efficacy of adenovirus-mediated gene transfer. Since the mouse myoblasts, C2C12 cells, showed low sensitivity to infection by recombinant adenovirus 5, we initially infected these cells at a high multiplicity of infection (MOI) of 250 with the recombinant adenovirus containing hCAR cDNA and LacZ gene. Subsequent infection by recombinant adenovirus containing the marker gene, green fluorescence protein, became efficient even at a low MOI of 25. Thus, elevated hCAR expression in mouse muscle fibers made a second virus inoculation at low doses possible. We also demonstrated that the elevated hCAR expression did not influence muscle membrane integrity. Our results suggest that co-expression of CAR and a therapeutic gene by adenovirus vector constitutes a novel strategy to advance gene therapy for hereditary muscle diseases. PMID- 11402299 TI - Polyethylenimine/DNA complexes shielded by transferrin target gene expression to tumors after systemic application. AB - Systemic application of positively charged polycation/DNA complexes has been shown to result in predominant gene expression in the lungs. Targeting gene expression to other sites, eg distant tumors, is hampered by nonspecific interactions largely due to the positive surface charge of transfection complexes. In the present study we show that the positive surface charge of PEI (25 kDa branched or 22 kDa linear)/DNA complexes can be efficiently shielded by covalently incorporating transferrin at sufficiently high densities in the complex, resulting in a dramatic decrease in nonspecific interactions, eg with erythrocytes, and decreased gene expression in the lung. Systemic application of transferrin-shielded PEI/DNA complexes into A/J mice bearing subcutaneously growing Neuro2a tumors via the tail vein resulted in preferential (100- to 500 fold higher) luciferase reporter gene expression in distant tumors as compared with the major organs including the lungs. Tumor targeting is also demonstrated by DNA uptake and beta-galactosidase gene expression in tumor cells. Assessing DNA distribution following systemic application significant amounts of DNA were found in the liver and tumor. However, in the liver, DNA was mainly taken up by Kupffer cells and degraded without significant transgene expression. In the tumor, DNA was associated mainly with tumor cells and frequently found near structures which resemble primitive blood vessels. PMID- 11402300 TI - Identification of polyamides that enhance adenovirus-mediated gene expression in the urothelium. AB - Adenovirus-mediated gene therapy of bladder diseases has been limited by the inability to transduce the urothelium successfully using adenoviral vectors. We have sought to identify agents that would increase adenovirus-mediated transgene expression in the bladder. We have utilized a rat model to screen compounds for their ability to enhance viral transgene expression in the rat bladder. Rats received intravesical administration of replication-deficient adenovirus (rAd) formulated in various agents, and transgene expression was evaluated after 48 h by determining the amount of lacZ expression in the luminal epithelium of the bladder. We report the identification of two different polyamides, each capable of dramatically increasing viral transgene expression in the bladder without causing detectable alteration of the umbrella cell layer of the urothelium. We have utilized a carcinogen-induced rat bladder tumor model to demonstrate that these polyamides are also capable of enhancing viral transgene expression in tumor tissue. The identification of these polyamides potentiates the use of adenovirus-mediated gene therapy for the treatment of superficial bladder cancer or other bladder diseases. PMID- 11402301 TI - Influence of adenoviral fiber mutations on viral encapsidation, infectivity and in vivo tropism. AB - Targeting of adenovirus (Ad)-encoded therapeutic genes to specific cell types has become a major goal in gene therapy. Redirecting the specificity of infection requires the abrogation of the natural interaction between the viral fiber and its cellular receptors (CAR) and the simultaneous introduction of a new binding specificity into the viral capsid. To abrogate the natural affinity of the fiber, we have mutated residues presumed to be directly or indirectly involved in CAR binding in the knob domain of the fiber protein. These residues are located in the AB loop (Ser408) and in the DG loop (Tyr491, Ala494, Ala503). The mutations Ser408Glu, Tyr491Asp, Ala494Asp and Ala503Asp did not prevent the incorporation of trimeric fibers in the viral capsid but led to loss of CAR binding in vitro. Infectivity of the mutant viruses could be restored in vitro by introducing a ligand at the C-terminal end of the knob, confirming that the reduced infectivity of the fiber-modified virus was due to an impaired interaction of the viral particle with the CAR receptor. However, after systemic delivery, the in vivo biodistribution of impaired CAR-binding viruses without addition of a specific ligand was not altered when compared with wild-type Ad. PMID- 11402302 TI - A self-immunomodulating myoblast cell line for erythropoietin delivery. AB - The transplantation of genetically engineered cells faces limitations associated with host immunity. Allogeneic cells are typically rejected in response to inherent histo-incompatibility. Even autologous cells can induce an immune response toward antigenic molecules expressed following transfer of foreign genes. The goal of the present study was to investigate the ability of immunomodulating molecules co-expressed with biotherapeutic factors to overcome these limitations both in syngeneic and allogeneic cell transplantation. The C(2)C(12) mouse myoblast cell line was engineered to express CTLA4Ig, a soluble factor blocking T cell costimulation, in conjunction with erythropoietin (Epo), a reporter biotherapeutic protein. In syngeneic C3H mice, myoblasts expressing only mouse Epo were mostly rejected within 2 weeks, as indicated by the transient increase in host hematocrit. In allogeneic recipients, the same cells induced only a 1-week increase in the hematocrit reflecting an acute rejection process. CTLA4Ig expression significantly extended the survival of mouse Epo-secreting myoblasts in approximately half of syngeneic hosts, whereas it led only to a 1 week improvement effect in allogeneic recipients. When combined with a transient anti-CD154 treatment, CTLA4Ig expression prevented Epo-secreting C(2)C(12)myoblasts from being rejected in allogeneic DBA/2J recipients for at least 1 month. In contrast, the same anti-CD154 treatment alone induced only a 1 week improvement. These results demonstrate that CTLA4Ig co-expression associated with a transient anti-CD154 treatment can prolong the delivery of recombinant proteins via transfer of ex vivo modified cells in allogeneic recipients. PMID- 11402303 TI - Systemic gene therapy with anti-angiogenic factors inhibits spontaneous breast tumor growth and metastasis in MMTVneu transgenic mice. AB - Tumor growth and metastasis are angiogenesis-dependent. The possibility of inhibiting tumor growth by interfering with the formation of new vessels has recently raised considerable interest. We previously reported that it is possible to inhibit primary tumor growth and metastasis in a transgenic model of spontaneous breast tumor, which shows many similarities to its human counterpart (including ability to metastasize) by intratumoral administration of a DNA construct carrying the murine angiostatin cDNA driven by liposomes. Here we report that it is also possible to achieve this goal by a systemic (intraperitoneal) delivery of therapeutic DNA constructs carrying genes coding for mouse and human anti-angiogenic factors which include angiostatin, endostatin and TIMP-2. These findings may be relevant to the design of therapeutic interventions in humans. PMID- 11402304 TI - Efficient in vivo gene transfer by PCR amplified fragment with reduced inflammatory activity. AB - There are many problems associated with plasmid DNA that may limit its use in systemic gene transfer. These problems could be solved by the use of synthetic genes. As a model to test the feasibility of using synthetic genes for gene therapy, we PCR-amplified a fragment containing the CMV promoter, the luciferase gene and a polyadenylation signal. The in vivo expression efficiency of the PCR fragment was determined by using two different methods, a hydrodynamics-based gene transfer of naked DNA to the liver and LPD (a lipid-based vector) mediated gene transfer to the lung. Our results show that linear fragments are at least as active as plasmid DNA following systemic delivery by LPD. However, PCR fragments are much less inflammatory than plasmid DNA as shown by a three-fold reduction in serum levels of both TNF-alpha and IL-12. Our results also showed that PCR fragments are highly efficient in liver gene transfer following systemic administration in a large volume. Thus, these results support the idea of using synthetic genes for gene therapy. Since gene sequence can be easily obtained as a PCR fragment, our results also imply that it may provide a useful and convenient method for determining the physiologic function of a putative gene in intact animals. PMID- 11402305 TI - Prevention of spontaneous neu-expressing mammary tumor development in mice transgenic for rat proto-neu by DNA vaccination. AB - The HER-2/neu proto-oncogene is overexpressed in 20-30% of human breast cancers and is associated with high recurrence risk. The oncogenic potential of HER 2/neu, together with its elevated expression in tumors, cell surface localization, and immunogenicity in some patients, make this oncoprotein an ideal target for immunotherapeutic approaches. To test the efficacy of immune-based strategies in eliciting an antitumor response, we used the N#202 transgenic mouse model engineered to overexpress the rat neu proto-oncogene under the control of the mouse mammary tumor virus promoter; females of this line develop spontaneous focal mammary tumors by 6 months of age. Transgenic mice immunized intramuscularly with a HER-2 cDNA ligated into the VR1012 (VICAL) expression vector under the control of the cytomegalovirus promoter developed significantly fewer spontaneous tumors as compared with mice injected with the empty vector (P < 0.0001) or not injected (P = 0.0006). However, this protection was observed only when immunization was started in 3-month-old but not in 6-month-old mice. These data suggest that the xenogeneic HER-2 DNA sequence can break immune tolerance to rat neu in transgenic N#202 mice and induce protective immunity that impairs the neu oncogene-driven progression of mammary carcinogenesis. The preventive effect achieved by our immunological approach appeared not to be based on anti-neu specific B and T cell immune attacks but was more possibly based on different mechanisms including aspecific and inflammatory immunological responses. PMID- 11402306 TI - HVJ-liposome-mediated transfection of HSVtk gene driven by AFP promoter inhibits hepatic tumor growth of hepatocellular carcinoma in SCID mice. AB - Suicide gene therapy using ganciclovir (GCV) with transfection of the herpes thymidine kinase (HSVtk) gene has been studied for cancer therapy. The present study demonstrates an efficient method of suicide gene therapy for multiple hepatic tumors, involving repetitive transfection of the HSVtk gene driven by the alpha-fetoprotein (AFP) promoter using hemagglutinating virus of Japan (HVJ) liposomes. AFP-producing cells (HUH7) and AFP-nonproducing cells (LS180) were injected subcutaneously (s.c.) to establish tumors in nude mice. Two plasmid constructs, bacterial LacZ gene driven by the AFP promoter (AFPLacZ), and HSVtk gene driven by the AFP promoter (AFPTK1) were encapsulated into the HVJ-liposome and used. When AFPLacZ was injected into the s.c. tumors, expression of LacZ gene was confined to HUH7 tumors. Repeated transfection of AFPTK1 followed by GCV treatment markedly suppressed growth of HUH7 tumors, and apoptosis of HUH7 cells was recognized in the tumor. Next, HUH7 cells were injected into the portal vein in severe combined immunodeficiency mice to establish a hepatic tumor model. After inoculation with the tumor, HVJ-liposomes containing the AFPTK1 plasmid vector were injected into the portal vein via the splenic hilum, followed by GCV treatment. This gene therapy significantly inhibited the growth of tumors in the liver and markedly improved survival. Three injections of the AFPTK1 plasmid vector completely inhibited tumor growth. This procedure seems to have great potential for the treatment of multiple hepatic tumors. PMID- 11402307 TI - In vivo nuclear delivery of oligonucleotides via hybridizing bifunctional peptides. AB - Linking proteins directly to nucleic acids has been a complex task. By hybridizing a bifunctional peptide nucleic acid (PNA) consisting of a nucleic acid binding moiety and a nuclear localization signal (NLS) we have previously demonstrated that it is possible to link protein functions directly to nucleic acids containing a PNA target site. By hybridizing fluorescently labeled oligonucleotides to PNA-NLS molecules and subsequently transfecting different organs in vivo we demonstrate an active nuclear translocation of the PNA NLS/oligonucleotide complex in different mouse organs. PMID- 11402314 TI - Role of Gab proteins in phosphatidylinositol 3-kinase activation by thrombopoietin (Tpo). AB - In this study, we show that upon thrombopoietin (Tpo) stimulation the two adapter proteins Gab1 and Gab2 are strongly tyrosine phosphorylated and associated with Shc, SHP2, PI 3-kinase and Grb2 in mpl-expressing UT7 cells. Although Gab1 and Gab2 seem to mediate overlapping biological signals in many cells, only Gab1 is expressed and phosphorylated in response to Tpo in primary human megakaryocytic progenitors; furthermore, it associates with the same proteins. Although a low level of tyrosine phosphorylated IRS-2 protein is also detected in PI 3-kinase immunoprecipitates, Gab proteins are the essential proteins associated with PI 3 kinase after Tpo stimulation. We demonstrate that, albeit no association is detected between the Tpo receptor mpl and Gab proteins, Y112 located in the C terminal cytoplasmic domain of mpl is required for Gab1/2 tyrosine phosphorylation. Gab proteins are not tyrosine phosphorylated after Tpo stimulation of UT-7 and Ba/F3 cells expressing a mpl mutant lacking Y112. Moreover, no activation of the PI 3-kinase/Akt pathway is observed in cells expressing this mpl mutant. Finally, we show that this mutant does not allow cell proliferation, thereby confirming that PI 3-kinase activation is required for Tpo induced cell proliferation. PMID- 11402315 TI - Induction of the AP-1 members c-Jun and JunB by TGF-beta/Smad suppresses early Smad-driven gene activation. AB - Smad proteins transduce signals from TGF-beta receptors and regulate transcription of target genes. Among the latter are c-jun and junB, which encode members of the AP-1 family of transcription factors. In this study, we have investigated the functional interactions of the Smad and AP-1 transcription factors in the context of Smad-specific gene transactivation in both fibroblasts and keratinocytes. We demonstrate that overexpression of either junB or c-jun prevents TGF-beta- or Smad3-induced transactivation of the Smad-specific promoter construct (SBE)(4)-Lux. Inversely, Smad-driven promoter transactivation by TGF beta/Smad is significantly enhanced when c-jun expression is abolished in HaCaT keratinocytes, and when junB expression is prevented in fibroblasts, consistent with the cell-type specific induction of jun members by TGF-beta. We also demonstrate that Smad-specific gene transactivation in junB(-/-) mouse embryonic fibroblasts is significantly higher than in embryonic fibroblasts from the control parental mouse line, and that this difference is abolished by rescuing junB expression in junB(-/-) cells. Finally, we have determined that off-DNA interactions between Smad3 and both c-Jun and JunB result in the reduction of Smad3/DNA interactions. From these results, we provide a model in which jun expression in response to the initial Smad cascade represents a negative feed back mechanism counteracting Smad-driven gene transactivation. PMID- 11402316 TI - Efficient rejoining of radiation-induced DNA double-strand breaks in vertebrate cells deficient in genes of the RAD52 epistasis group. AB - Rejoining of ionizing radiation (IR) induced DNA DSBs usually follows biphasic kinetics with a fast (t(50): 5-30 min) component attributed to DNA-PK-dependent non-homologous endjoining (NHEJ) and a slow (t(50): 1-20 h), as of yet uncharacterized, component. To examine whether homologous recombination (HR) contributes to DNA DSB rejoining, a systematic genetic study was undertaken using the hyper-recombinogenic DT40 chicken cell line and a series of mutants defective in HR. We show that DT40 cells rejoin IR-induced DNA DSBs with half times of 13 min and 4.5 h and contributions by the fast (78%) and the slow (22%) components similar to those of other vertebrate cells with 1000-fold lower levels of HR. We also show that deletion of RAD51B, RAD52 and RAD54 leaves unchanged the rejoining half times and the contribution of the slow component, as does also a conditional knock out mutant of RAD51. A significant reduction (to 37%) in the contribution of the fast component is observed in Ku70(-/-) DT40 cells, but the slow component, operating with a half time of 18.4 h, is still able to rejoin the majority (63%) of DSBs. A double mutant Ku70(-/-)/RAD54(-/-) shows similar half times to Ku70(-/-) cells. Thus, variations in HR by several orders of magnitude leave unchanged the kinetics of rejoining of DNA DSBs, and fail to modify the contribution of the slow component in a way compatible with a dependence on HR. We propose that, in contrast to yeast, cells of vertebrates are 'hard-wired' in the utilization of NHEJ as the main pathway for rejoining of IR-induced DNA DSBs and speculate that the contribution of homologous recombination repair (HRR) is at a stage after the initial rejoining. PMID- 11402317 TI - DNA microarrays identification of primary and secondary target genes regulated by p53. AB - The transcriptional program regulated by the tumor suppressor p53 was analysed using oligonucleotide microarrays. A human lung cancer cell line that expresses the temperature sensitive murine p53 was utilized to quantitate mRNA levels of various genes at different time points after shifting the temperature to 32 degrees C. Inhibition of protein synthesis by cycloheximide (CHX) was used to distinguish between primary and secondary target genes regulated by p53. In the absence of CHX, 259 and 125 genes were up or down-regulated respectively; only 38 and 24 of these genes were up and down-regulated by p53 also in the presence of CHX and are considered primary targets in this cell line. Cluster analysis of these data using the super paramagnetic clustering (SPC) algorithm demonstrate that the primary genes can be distinguished as a single cluster among a large pool of p53 regulated genes. This procedure identified additional genes that co cluster with the primary targets and can also be classified as such genes. In addition to cell cycle (e.g. p21, TGF-beta, Cyclin E) and apoptosis (e.g. Fas, Bak, IAP) related genes, the primary targets of p53 include genes involved in many aspects of cell function, including cell adhesion (e.g. Thymosin, Smoothelin), signaling (e.g. H-Ras, Diacylglycerol kinase), transcription (e.g. ATF3, LISCH7), neuronal growth (e.g. Ninjurin, NSCL2) and DNA repair (e.g. BTG2, DDB2). The results suggest that p53 activates concerted opposing signals and exerts its effect through a diverse network of transcriptional changes that collectively alter the cell phenotype in response to stress. PMID- 11402318 TI - Multiple oral squamous epithelial lesions: are they genetically related? AB - The development of second primary tumors (SPTs) in patients with head and neck squamous cell carcinoma (HNSCC) has become an increasingly important factor in clinical treatment decisions. Currently, clinical and histologic parameters are used to determine whether or not SPT is present. Recent studies suggest that many SPTs in the upper aerodigestive tract have a common clonal origin, challenging the longstanding multiclonal origin concept. To determine genetic relationships among multiple oral cancerous and precancerous lesions (MOCP), we analysed 100 lesions from 26 Japanese patients. Lesion development was synchronous and metachronous. We looked for patterns of microsatellite alterations (MA) using seven markers at chromosomes 3p14, 9p21, and 17p13, where MA occurs early in oral carcinogenesis. Loss of heterozygosity (LOH) was found in 52.6% (41/78), 62.5% (60/96), and 59.3% (32/54) of informative MOCP at 3p14, 9p21, and 17p13, respectively. Microsatellite instability (MI) was observed in 11, 26 and 13% of the samples at 3p14, 9p21, and 17p13 markers, respectively. Patterns of MA were concordant in only nine (14%) of 63 lesions from four (18%) of 22 patients who initially presented with noninvasive lesions. However, two of four patients with invasive cancer as indexed lesion showed 16 (43%) clonally related MOCP among 37 lesions (P=0.003). The results suggest that the majority of MOCP arise from clonally independent cells affected by field cancerization. However, the probability of mucosal spread of clonal malignant or premalignant cells may increase along with malignant progression. PMID- 11402319 TI - Expression of ring finger-deleted TRAF2 sensitizes metastatic melanoma cells to apoptosis via up-regulation of p38, TNFalpha and suppression of NF-kappaB activities. AB - Mechanisms underlying radiation and chemotherapy resistance, the hallmark of human melanoma, are not well understood. Here we demonstrate that expression levels of signal adaptor protein TRAF2 coincide with melanoma resistance to UV irradiation. Altered TRAF2 signaling by a form of TRAF2, which lacks the ring finger domain (TRAF2DeltaN), increases activities of p38 MAPK, ATF2, and the level of TNFalpha expression. Forced expression of TRAF2DeltaN in HHMSX highly metastatic melanoma cells that lack Fas expression and thus utilize the TNFalpha TNFR1 as the major apoptotic pathway sensitized cells to UV-induced apoptosis. An over twofold increase in degree of apoptosis was observed in TRAF2DeltaN expressing cells that were treated with actinomycin D, anisomycin or with the radiomimetic drug neocarzinostatin. Sensitization by TRAF2DeltaN is selective since it was not observed in response to either Taxol or cis-platinum treatment. TRAF2DeltaN effects are primarily mediated via p38 since inhibition of p38 reduces, whereas activation of p38 promotes the level of UV-induced apoptosis. Conversely, activation of IKK attenuates the sensitization of melanoma by TRAF2DeltaN, indicating that p38-mediated suppression of NF-kappaB activity is among TRAF2DeltaN effects. Our finding identifies p38, TNFalpha and NF-kappaB among key players that efficiently sensitizes melanoma cells to UV-, ribotoxic (anisomycin) and radiomimetic chemicals-induced programmed cell death in response to aberrant TRAF2 signaling. PMID- 11402320 TI - Inhibition of anchorage-independent growth and lung metastasis of A549 lung carcinoma cells by IkappaBbeta. AB - To evaluate the role of the NF-kappaB signaling pathway in oncogenic transformation, we expressed IkappaBbeta, a specific inhibitor of NF-kappaB, in two human lung adenocarcinoma cell lines, A549 and H441. Expression of IkappaBbeta significantly reduced NF-kappaB activation induced by cotransfection with p65/RelA or TNF-alpha and abrogated the basal NF-kappaB activity in A549 cells. Transfection of IkappaBbeta into A549, H441 and K-ras-transformed NIH3T3 cells suppressed anchorage-independent growth as measured by colony formation in soft agar. Anchorage-independent growth of vector-transfected A549 cells in reduced serum could be enhanced by both EGF and IGF-I. In contrast, only EGF but not IGF-I could induce anchorage-independent growth of IkappaBbeta-expressing A549 cells, suggesting that the IGF-I signaling pathway regulating growth and survival may be blocked by IkappaBbeta. Interestingly, expression of IkappaBbeta suppressed growth of A549 cells in low serum in vitro without affecting in vivo growth subcutaneously in nude mice. However, metastatic growth of IkappaBbeta expressing A549 cells in the lungs of nude mice was significantly inhibited. These results provide evidence that NFkappaB plays an important role in anchorage independent growth and metastatic growth of lung carcinoma cells. PMID- 11402321 TI - Reducing mammary cancer risk through premature stem cell senescence. AB - The reproductive capacity of the mammary epithelial stem cell is reduced coincident with the number of symmetric divisions it must perform. In a study of FVB/N mice with the transgene, WAP-TGFbeta1, we discovered that mammary epithelial stem cells were prematurely aged due to ectopic expression of TGF beta1. To test whether premature aging of mammary epithelial stem cells would have an impact on susceptibility or resistance to mammary cancer, female littermates from FVB/N x WAP-TGF-beta1 mating were injected with mouse mammary tumor virus (MMTV) at 8-10 weeks of age. A total of 44 females were inoculated, maintained as breeders and observed for tumor development for up to 18 months. Only one mammary tumor appeared in 17 TGF-beta1 females while 15 were collected from 29 wild type sisters. Premalignant mammary epithelial cells in infected glands were identified by transplantation of single cell (1 x 10(5)) suspensions into nulliparous hosts and testing for hyperplastic outgrowth. Although the number of positive takes was significantly reduced with TGF-beta1 cells, both MMTV-infected TGF-beta1 and wild type cells produced hyperplastic outgrowths suggesting that premalignant transformation was achieved in each group. The results suggest a positive correlation between the procreative life-span of mammary epithelial stem cells and mammary cancer risk. PMID- 11402322 TI - Limiting the location of putative human prostate cancer tumor suppressor genes on chromosome 18q. AB - We studied loss of heterozygosity (LOH) on the long arm of human chromosome 18 in prostate cancer to determine the location of a putative tumor suppressor gene (TSG) and to correlate these losses with the pathological grade and stage of the cancer. Of 48 specimens analysed 17 (35.4%) lost at least one allele on chromosome 18q. All the specimens with allelic losses lost at least one allele within chromosomal region 18q21. Allelic losses picked at D18S51 (19%) and D18S858 (17%). A 0.58 cM DNA segment that includes the D18S858 locus and is flanked by the microsatellite loci D18S41 and D18S381, was lost in eight (47%) of 17 specimens with allelic losses. This segment was designated as a LOH cluster region 1 (LCR 1). Although Smad2 resides within LCR 1, it was not mutated in any of the six prostate cell lines (five prostate cancer cell lines and one immortalized prostate epithelial cell line) analysed, suggesting that it is not a candidate TSG in prostate cancer. A second LCR at 18q21, LCR 2, includes the D18S51 locus and is flanked by the D18S1109 and D18S68 loci, which are separated by 7.64 cM. LCR 2 was lost in six (35%) of the 17 specimens with chromosome 18q losses. These results suggest that chromosome 18q21 may harbor two candidate prostate cancer TSGs. The candidate TSGs DCC and Smad4 are located centromeric to the LCRs. No alleles were lost within or in close proximity to these genes, suggesting that they are not targets for inactivation by allelic losses in prostate cancer. Although there was no obvious correlation between chromosome 18q LOH and the pathological grade or stage, three (37.5%) of eight low-grade cancers and nine (32.1%) of 28 organ-confined cancers lost alleles at 18q21, suggesting that allelic losses are relatively early events in the development of invasive prostate cancer. PMID- 11402323 TI - Immediate early genes induced by H-Ras in thyroid cells. AB - Expression of oncogenic v-H-Ras in the thyroid cell line FRTL-5 (FRTL-5(Ras)) results in uncontrolled proliferation, loss of thyroid-specific gene expression and tumorigenicity. Concomitant expression of constitutively activated MEK and Rac, two major H-Ras downstream effectors, in FRTL-5 (FRTL-5(MEK/Rac)) recapitulates H-Ras effects on proliferation and morphology. In contrast to FRTL 5(Ras), however, FRTL-5(MEK/Rac) cells remain differentiated and are not tumorigenic. To find H-Ras induced genes potentially responsible for tumorigenicity and loss of differentiation, we have used subtractive suppression hybridization (SSH), a PCR-based cDNA subtraction technique, between de differentiated and tumorigenic FRTL-5(Ras) cells and differentiated and non tumorigenic FRTL-5(MEK/Rac) cells. We examined 800 of the cDNA clones obtained after subtraction and verified their levels of expression in the two cell lines by reverse northern, identifying 337 H-Ras induced genes. By sequence analysis, we clustered 57 different genes. Among these, 39 were known genes (involved in diverse signal transduction processes regulating mitogenic activity, cell survival, cytoskeletal reorganization, stress response and invasion) while the remaining 18 clones were novel genes. Among the 57 H-Ras specific clones, we identified those genes whose expression is induced early by H-Ras. We suggest that these immediate-early genes may play a crucial role in H-Ras-mediated transformation in thyroid epithelial cells. PMID- 11402324 TI - Bcl-2 expression in F-MuLV-induced erythroleukemias: a role for the anti apoptotic action of Bcl-2 during tumor progression. AB - Erythroleukemias induced by various strains of Friend virus are multistage malignancies that result from the accumulation of genetic mutations, including the activation of proto-oncogenes and the inactivation of tumor suppressor genes. In this study, we demonstrate that Bcl-2 expression is activated in the majority of F-MuLV-induced erythroleukemia cell lines. In contrast, Bcl-2 was not expressed in any of the FV-P-induced erythroleukemia cell lines and protein levels were low or negligible in FV-A-induced erythroleukemia cell lines examined. In vivo, Bcl-2 expression levels gradually increased in F-MuLV-induced erythroleukemic cells prior to adaptation to culture. High expression of Bcl-2 in F-MuLV-induced erythroleukemic cells was shown to proceed the emergence of p53 mutation suggesting that Bcl-2 expression may delay p53 mutation in the leukemic cells. This is further supported by the demonstration that the majority of F-MuLV induced erythroleukemia cell lines established from primary tumors induced in p53 mutant mice express low to negligible levels of Bcl-2. We have shown that the high levels of Bcl-2 expression in FV-P-induced erythroleukemic cells inhibited apoptosis induced by etoposide, low serum and p53 expression. Similarly, ectopic Bcl-2 expression within these cells also provided protection from apoptosis induced by etoposide and growth in low serum. These results suggest that the anti apoptotic action of Bcl-2 may confer a selective in vivo and in vitro growth advantage to F-MuLV-induced erythroleukemic cells, which is not shared by FV-P/FV A-induced erythroleukemic cells. The observed induction of Bcl-2 expression in vivo constitutes a novel but late oncogenic event associated with the progression of F-MuLV-induced erythroleukemias. PMID- 11402325 TI - The constitutive activation of the CEF-4/9E3 chemokine gene depends on C/EBPbeta in v-src transformed chicken embryo fibroblasts. AB - The CEF-4/9E3 chemokine gene is expressed constitutively in chicken embryo fibroblasts (CEF) transformed by the Rous sarcoma virus (RSV). This aberrant induction is controlled at the transcriptional and post-transcriptional levels. Transcriptional activation depends on multiple elements of the CEF-4 promoter composing a Src-responsive-Unit or SRU. The SRU includes a TPA responsive element, a PRDII/kappaB domain and a CAAT box. In this report, we identify C/EBPbeta as a component of the trans-acting factor interacting with the CAAT box of the CEF-4 promoter. In addition, we show that C/EBPbeta binds to a second element located in proximity of the TRE. A mutation of this distal CAAT box impaired the activation of the CEF-4 promoter by pp60(v-src) indicating that this element is also part of the SRU. Using the RCASBP retroviral vector, we expressed a dominant negative mutant of C/EBPbeta (designated Delta184-C/EBPbeta) in RSV transformed CEF. Delta184-C/EBPbeta decreased the accumulation of the CEF-4 mRNA and activation of the CEF-4 promoter by pp60(v-src). The induction of the Cox-2 gene (CEF-147) was also reduced by Delta184-C/EBPbeta. The effect of the dominant negative mutant was observed within 1 h of the activation of a thermolabile pp60(v-src) suggesting that C/EBPbeta is an early target of v-src transformation. The dominant negative mutant did not inhibit the transformation of CEF by RSV and in fact accentuated the transformed cell phenotype. Therefore, the activation of C/EBPbeta is important for the expression of v-src regulated genes but is not required for the in vitro transformation of CEF by pp60(v-src). PMID- 11402326 TI - In vitro and in vivo inhibition of epidermal growth factor receptor-tyrosine kinase pathway by photodynamic therapy. AB - PDT, a new therapeutic procedure for the management of many malignant conditions including skin cancer, involves the administration of a photosensitizing compound followed by illumination of the lesion with visible light. We earlier showed an involvement of: (i) WAF1/p21-cyclins (D1 and E)-cdk (2 and 6) network; and (ii) Rb/E2F-DP machinery during silicon phthalocyanine (Pc4)-PDT-mediated cell cycle dysregulation and apoptosis of human epidermoid carcinoma (A431) cells. Here, we investigated the involvement of EGFR-pathway during antiproliferative responses of Pc4-PDT in A431 cells and during ablation of murine skin papillomas. Pc4-PDT of A431 cells was found to result in a time-dependent down-modulation of the protein expression and phosphorylation of EGFR and Shc (an immediate downstream molecule in EGFR-pathway), during progressive increase in apoptotic response. To establish the relevance of these in vitro findings to in vivo situations, we subjected chemically- as well as ultraviolet B radiation-induced squamous papillomas in SENCAR and SKH-1 hairless mice, respectively, to Pc4-PDT, and assessed its effect on EGFR-pathway during ablation of these tumors. Pc4-PDT was found to result in a time-dependent: (i) inhibition of protein expressions of EGFR; and (ii) tyrosine phosphorylation of EGFR and Shc; and (iii) induction of apoptosis, during the regression of these tumors. These data suggest the involvement of EGFR-pathway during the antiproliferative effects of PDT. It is tempting to speculate that inhibitors of EGFR could enhance the therapeutic efficacy of PDT. PMID- 11402327 TI - The 'wildtype' conformation of p53: epitope mapping using hybrid proteins. AB - The function of p53 correlates with its 'wildtype' conformation, specifically recognized by antibodies PAb1620 and PAb246, and many cancer-associated mutations cause loss of this conformation. The epitopes of these antibodies were identified using hybrid p53 proteins created by a new method. Plasmids carrying homologous genes cut at appropriate sites recombined efficiently when transformed into RecE(+) E. coli. PAb1620 and PAb246 recognize mouse but not chicken p53; we created mouse-chicken hybrids of the p53 core domain and tested antibody reactivity. PAb246 binding mapped to residues 201-212, while PAb1620 required both residues 145-157 and 201-212 (human p53 numbering used throughout). An alanine-scan showed that the key residues for PAb246 and PAb1620 are completely distinct: PAb246 recognizes residues 202-204 (Tyr-Pro-Glu) while PAb1620 recognizes residues Arg156, Leu206, Arg209, and Gln/Asn210, the last two residues being essential. Both antibody epitopes are far from the p53 interface with DNA, but near the epitope of the 'mutant' conformation antibody PAb240. These epitope locations may help in dissecting the interactions of p53, including those with E6/E6-AP and in its DNA-bound state. PMID- 11402328 TI - The SV40 small t-antigen prevents mammary gland differentiation and induces breast cancer formation in transgenic mice; truncated large T-antigen molecules harboring the intact p53 and pRb binding region do not have this effect. AB - We report here for the first time, that the SV40 small t-antigen inhibits mammary gland differentiation during mid-pregnancy and that about 10% of multiparous WAP SVt transgenic animals develop breast tumors with latencies ranging from 10-17 months. Cyclin D1 is deregulated and over expressed in the small t-antigen positive mammary gland epithelial cells (ME-cells) and in the breast tumor cells. SV40 small t-antigen immortalized ME-cells (t-ME-cells) exhibit a strong intranuclear cyclin D1 staining, also in the absence of external growth factors and the cells continue to divide for several days without serum. In addition, the expression rate of cyclin E and p21(Waf1) but not of p53 is increased. Coimmunoprecipitation experiments revealed that p21(Waf1) is mainly associated with the cyclin D/CDK4 but not with the cyclin E/CDK2 complex. WAP-SVT transgenic animals exhibit an almost regular mammary gland development until late pregnancy but the majority of the ME-cells are eliminated by apoptosis during the early lactation period. Tumor formation is delayed and less efficient than in T/t antigen positive animals. Sequestration of p53 and pRb by the N-terminal truncated T-antigen molecules (T1-antigen and T2-antigen) does not affect mammary gland differentiation and the transgenic animals (WAP-SVBst-Bam) do not develop breast tumors. PMID- 11402330 TI - AP-1--Introductory remarks. AB - This issue attempts to give a 'state of the art' overview of the AP-1 transcription factor family, a fundamental class of transcriptional regulators. The AP-1 family consists of several bZIP (basic region leucine zipper) domain proteins, the Jun, the Fos, and the ATF subfamilies, which all have to dimerize before they can bind to their DNA target sites. AP-1 has been a paradigm for transcription factors that regulate many aspects of cell physiology in response to environmental changes such as stress, radiation, or to growth factor signals thereby acting like an environmental biosensor. Although we have come a long way from discovering its major components, the heterodimer between c-Fos and c-Jun, it is daunting to realize that we still lack a detailed molecular knowledge of how these factors interact with DNA to activate or repress genes in the nucleus. It is also not clear how the response of AP-1 to growth factor signaling from the cell surface to the nucleus is interpreted at the molecular level and whether AP 1 is relevant for human disease. PMID- 11402331 TI - Redox control of AP-1-like factors in yeast and beyond. AB - Cells have evolved complex and efficient strategies for dealing with variable and often-harsh environments. A key aspect of these stress responses is the transcriptional activation of genes encoding defense and repair proteins. In yeast members of the AP-1 family of proteins are required for the transcriptional response to oxidative stress. This sub-family of AP-1 (called yAP-1) proteins are sensors of the redox-state of the cell and are activated directly by oxidative stress conditions. yAP-1 proteins are bZIP-containing factors that share homology to the mammalian AP-1 factor complex and bind to very similar DNA sequence sites. The generation of reactive oxygen species and the resulting potential for oxidative stress is common to all aerobically growing organisms. Furthermore, many of the features of this response appear to be evolutionarily conserved and consequently the study of model organisms, such as yeast, will have widespread utility. The important structural features of these factors, signaling pathways controlling their activity and the nature of the target genes they control will be discussed. PMID- 11402332 TI - Drosophila AP-1: lessons from an invertebrate. AB - In recent years, studies in the model organism Drosophila melanogaster have contributed significant insights into the molecular and developmental biology of the AP-1 transcription factors Jun and Fos. Powerful genetic and biochemical approaches uncovered a baffling complexity and variability of the signaling connections to and from AP-1. The range of biological processes that Jun and Fos regulate in this organism is equally multi-faceted. Regulatory interactions between AP-1 and JNK, ERK, TGFbeta, Notch or other signaling systems have been implicated in the control of a multitude of embryonic and adult events, including tissue closure processes, patterning of eye, gut and wing, as well as apoptosis. Here we review the information that has been gathered on Drosophila AP-1 in signal transduction and on the developmental and cellular functions controlled by AP-1-mediated signals in the fly. Lessons learned from the studies on AP-1 in Drosophila may contribute to our general understanding, beyond species boundaries, of this fundamental class of transcriptional regulators. PMID- 11402333 TI - Jun, the oncoprotein. AB - Cellular Jun (c-Jun) and viral Jun (v-Jun) can induce oncogenic transformation. For this activity, c-Jun requires an upstream signal, delivered by the Jun N terminal kinase (JNK). v-Jun does not interact with JNK; it is autonomous and constitutively active. v-Jun and c-Jun address overlapping but not identical sets of genes. Whether all genes essential for transformation reside within the overlap of the v-Jun and c-Jun target spectra remains to be determined. The search for transformation-relevant targets of Jun is moving into a new stage with the application of DNA microarrays technology. Genetic screens and functional tests remain a necessity for the identification of genes that control the oncogenic phenotype. PMID- 11402334 TI - The mammalian Jun proteins: redundancy and specificity. AB - The AP-1 transcription factor is composed of a mixture of homo- and hetero-dimers formed between Jun and Fos proteins. The different Jun and Fos family members vary significantly in their relative abundance and their interactions with additional proteins generating a complex network of transcriptional regulators. Thus, the functional activity of AP-1 in any given cell depends on the relative amount of specific Jun/Fos proteins which are expressed, as well as other potential interacting proteins. This diversity of AP-1 components has complicated our understanding of AP-1 function and resulted in a paucity of information about the precise role of individual AP-1 members in distinct cellular processes. We shall discuss recent studies which suggest that different Jun and Fos family members may have both opposite and overlapping functions in cellular proliferation and cell fate. PMID- 11402335 TI - AP-1 in cell proliferation and survival. AB - A plethora of physiological and pathological stimuli induce and activate a group of DNA binding proteins that form AP-1 dimers. These proteins include the Jun, Fos and ATF subgroups of transcription factors. Recent studies using cells and mice deficient in individual AP-1 proteins have begun to shed light on their physiological functions in the control of cell proliferation, neoplastic transformation and apoptosis. Above all such studies have identified some of the target genes that mediate the effects of AP-1 proteins on cell proliferation and death. There is evidence that AP-1 proteins, mostly those that belong to the Jun group, control cell life and death through their ability to regulate the expression and function of cell cycle regulators such as Cyclin D1, p53, p21(cip1/waf1), p19(ARF) and p16. Amongst the Jun proteins, c-Jun is unique in its ability to positively regulate cell proliferation through the repression of tumor suppressor gene expression and function, and induction of cyclin D1 transcription. These actions are antagonized by JunB, which upregulates tumor suppressor genes and represses cyclin D1. An especially important target for AP-1 effects on cell life and death is the tumor suppressor p53, whose expression as well as transcriptional activity, are modulated by AP-1 proteins. PMID- 11402336 TI - AP-1 in mouse development and tumorigenesis. AB - Genetically modified mice have provided important insights into the biological functions of the dimeric transcription factor complex AP-1. Extensive analyses of mice and cells with genetically modified Fos or Jun proteins provide novel insights into the physiological functions of AP-1 proteins. Using knock-out strategies it was found that some components, such as c-Fos, FosB and JunD are dispensable, whereas others, like c-Jun, JunB and Fra-1 are essential in embryonic development and/or in the adult organism. Besides the specific roles of AP-1 proteins in developmental processes, we are beginning to obtain a better molecular understanding of the cell-context dependent function of AP-1 in cell proliferation and apoptosis, in bone biology as well as in multistep tumorigenesis. PMID- 11402337 TI - Function and regulation of AP-1 subunits in skin physiology and pathology. AB - The mouse skin has become the model of choice to study the regulation and function of AP-1 subunits in many physiological and pathological processes in vivo and in vitro. Genetically modified mice, in vitro reconstituted skin equivalents and epidermal cell lines were established, in which AP-1-regulated genetic programs of cell proliferation, differentiation and tumorigenesis can be analysed. Since the epidermis, as our interface with the environment, is subjected to radiation and injury, signal transduction pathways and critical AP-1 members regulating the mammalian stress response could be identified. Regulated expression of important components of the cytokine network, cell surface receptors and proteases, which orchestrate the process of wound healing has been found to rely on AP-1 activity. Here we review our current knowledge on the function of AP-1 subunits and AP-1 target genes in these fascinating fields of skin physiology and pathology. PMID- 11402338 TI - AP-1 proteins in the adult brain: facts and fiction about effectors of neuroprotection and neurodegeneration. AB - Jun and Fos proteins are induced and activated following most physiological and pathophysiological stimuli in the brain. Only few data allow conclusions about distinct functions of AP-1 proteins in neurodegeneration and neuroregeneration, and these functions mainly refer to c-Jun and its activation by JNKs. Apoptotic functions of activated c-Jun affect hippocampal, nigral and primary cultured neurons following excitotoxic stimulation and destruction of the neuron-target axis including withdrawal of trophic molecules. The inhibition of JNKs might exert neuroprotection by subsequent omission of c-Jun activation. Besides endogenous neuronal functions, the c-Jun/AP-1 proteins can damage the nervous system by upregulation of harmful programs in non-neuronal cells (e.g. microglia) with release of neurodegenerative molecules. In contrast, the differentiation with neurite extension and maturation of neural cells in vitro indicate physiological and potentially neuroprotective functions of c-Jun and JNKs including sensoring for alterations in the cytoskeleton. This review summarizes the multiple molecular interfunctions which are involved in the shift from the physiological role to degenerative effects of the Jun/JNK-axis such as cell type specific expression and intracellular localization of scaffold proteins and upstream activators, antagonistic phosphatases, interaction with other kinase systems, or the activation of transcription factors competing for binding to JNK proteins and AP-1 DNA elements. PMID- 11402339 TI - Close encounters of many kinds: Fos-Jun interactions that mediate transcription regulatory specificity. AB - Fos and Jun family proteins regulate the expression of a myriad of genes in a variety of tissues and cell types. This functional versatility emerges from their interactions with related bZIP proteins and with structurally unrelated transcription factors. These interactions at composite regulatory elements produce nucleoprotein complexes with high sequence-specificity and regulatory selectivity. Several general principles including binding cooperativity and conformational adaptability have emerged from studies of regulatory complexes containing Fos-Jun family proteins. The structural properties of Fos-Jun family proteins including opposite orientations of heterodimer binding and the ability to bend DNA can contribute to the assembly and functions of such complexes. The cooperative recruitment of transcription factors, coactivators and chromatin remodeling factors to promoter and enhancer regions generates multiprotein transcription regulatory complexes with cell- and stimulus-specific transcriptional activities. The gene-specific architecture of these complexes can mediate the selective control of transcriptional activity. PMID- 11402340 TI - Distinct roles of Jun : Fos and Jun : ATF dimers in oncogenesis. AB - Jun : Fos and Jun : ATF complexes represent two classes of AP-1 dimers that (1) preferentially bind to either heptameric or octameric AP-1 binding sites, and (2) are differently regulated by cellular signaling pathways and oncogene products. To discriminate between the functions of Jun : Fos, Jun : ATF and Jun : Jun, mutants were developed that restrict the ability of Jun to dimerize either to itself, or to Fos(-like) or ATF(-like) partners. Introduction of these mutants in chicken embryo fibroblasts shows that Jun : Fra2 and Jun : ATF2 dimers play distinct, complementary roles in in vitro oncogenesis by inducing either anchorage independence or growth factor independence, respectively. v-Jun : ATF2 rather than v-Jun : Fra2 triggers the development of primary fibrosarcomas in the chicken wing. Genes encoding extracellular matrix components seem to constitute an important subset of v-Jun : ATF2-target genes. Repression of the matrix component SPARC by Jun is essential for the induction of fibrosarcomas. Avian primary cells transformed by either Jun : Fra2 or Jun : ATF2 thus provide powerful tools for the investigation of the downstream pathways involved in oncogenesis. Further genetic studies with Jun dimerization mutants will be required to be precise and extend the specific roles of the Jun : Fos and Jun : ATF dimers during cancer progression in avian and mammalian systems. PMID- 11402341 TI - Cross-talk between glucocorticoid receptor and AP-1. AB - Cross-talk between different transcription factors, notably between the glucocorticoid receptor and AP-1, has been discovered more than 10 years ago: a bona fide transcription factor, without apparent need for its own direct DNA contact, influences the activity of another transcription factor. Recent experiments have added interesting aspects: in addition to major insights into the mechanism of cross-talk, it is now clear that the cross-talk ability of glucocorticoid receptor is essential for mouse development, while the activation of target promoters carrying a glucocorticoid response element (GRE), is surprisingly, dispensable for survival under animal house conditions. Interestingly, the cross-talk function is responsible for almost all regulatory actions of cortisol in the immune system. It is possible that the two functions of the glucocorticoid receptor can be activated separately by specific ligands. Future goals will be to define whether adverse effects of long-term corticosteroid treatment, e.g. osteoporosis, joint necroses, metabolic effects, can be ascribed to GRE-target gene activation and thus be dissociated from the desirable actions in the treatment e.g. of autoimmune disease. PMID- 11402342 TI - Partners in transcription: NFAT and AP-1. AB - Combinatorial regulation is a powerful mechanism that enables tight control of gene expression, via integration of multiple signaling pathways that induce different transcription factors required for enhanceosome assembly. The four calcium-regulated transcription factors of the NFAT family act synergistically with AP-1 (Fos/Jun) proteins on composite DNA elements which contain adjacent NFAT and AP-1 binding sites, where they form highly stable ternary complexes to regulate the expression of diverse inducible genes. Concomitant induction of NFAT and AP-1 requires concerted activation of two different signaling pathways: calcium/calcineurin, which promotes NFAT dephosphorylation, nuclear translocation and activation; and protein kinase C (PKC)/Ras, which promotes the synthesis, phosphorylation and activation of members of the Fos and Jun families of transcription factors. A fifth member of the NFAT family, NFAT5, controls the cellular response to osmotic stress, by a mechanism that requires dimer formation and is independent of calcineurin or of interaction with AP-1. Pharmacological interference with theNFAT:AP-1 interaction may be useful in selective manipulation of the immune response. Balanced activation of NFAT and AP-1 is known to be required for productive immune responses, but the role of NFAT:AP-1 interactions in other cell types and biological processes remains to be understood. PMID- 11402343 TI - Signal transduction by MAP kinases in T lymphocytes. PMID- 11402344 TI - Preparing for the worst. PMID- 11402345 TI - The long march to primary health care in China: from collectivism to market economics. PMID- 11402346 TI - The incidence and aetiology of stroke in the Caerphilly and Speedwell Collaborative Studies I: methods and incidence of events. AB - Stroke mortality and morbidity remain high despite downward trends in incidence and case fatality. Population-based longitudinal studies which include collection of risk factor data are required for a better understanding of stroke aetiology. From a representative cohort of men from South Wales and South-west England, followed up for a median of 17 y, details of possible cerebrovascular events were collected from questionnaires, hospital admission data, general practitioner records, death certificates, radiology records and post-mortem reports. Radiology records, and strokes and transient ischaemic attacks were independently validated. There were 433 strokes and 163 transient ischaemic attacks identified during follow-up. Of these, 333 were the first ever in a lifetime strokes of which 139 were definite ischaemic, 20 were haemorrhagic and 168 were probable ischaemic strokes. The crude incidence rate for stroke was 445 (95% confidence interval 398-493) per 100 000 person years. The age-standardised rates for 10 y age-bands were: 45-54 y 91 (10-172); 55-64 y 351 (269-432) and 65-74 y 855 (669 1040). The 30 d case-fatality rate was 21.0% (70/333) for all strokes and 19.2% (60/312) for ischaemic strokes. For transient ischaemic attacks the age standardised incidence rates for the same 10 y age bands were 92 (4-179), 111 (64 157), and 273 (167-80), respectively. These rates for stroke transient ischaemic attack are likely to be accurate given the high ascertainment of events in this representative population of middle-aged men. Such studies, reporting reliable measures of cerebrovascular events, are important for measuring burden of disease, and for analysis of risk factor associations to help improve understanding of stroke aetiology and inform preventive efforts. PMID- 11402347 TI - The incidence and aetiology of stroke in the Caerphilly and Speedwell Collaborative Studies II: risk factors for ischaemic stroke. AB - Reduction of stroke burden requires preventive interventions targeted at important risk factors. This report presents the analysis of risk factors for ischaemic stroke from a representative cohort of middle aged men from South Wales and south-west England. Data on risk factors were collected through validated questionnaires and physical and clinical measurements. Details of possible cerebrovascular events were retrieved, classified into ischaemic, haemorrhagic and uncertain subtypes, and validated. The ratio of definite ischaemic to definite haemorrhagic strokes was calculated. This showed that the vast majority of strokes of unknown subtype were likely to ischaemic. After exclusion of known haemorrhagic strokes and subarrachnoid haemorrhages the remaining strokes were labelled ischaemic. Hazard ratios for possible risk factors were calculated for all ischaemic, and for fatal and non-fatal strokes. There were 293 ischaemic strokes. Statistically significant age-adjusted hazard ratios were: 1.50 (95% confidence interval 1.16-1.95) for being in a manual social class, 1.82 (1.24 2.67) if smoking >15 cigarettes/d at enrollment, 1.19 (1.13-1.24) and 1.23 (1.14 1.34) per 10 mmHg increase in systolic and diastolic blood pressure, respectively, 0.67 (0.46-0.96) for the top quintile high density lipoprotein cholesterol:cholesterol ratio compared to the bottom quintile, 2.04 (1.40-2.99) for presence of angina, 3.90 (2.01-7.58) for presence of atrial fibrillation, and 3.35 (1.90-5.80) for presence of diabetes. Risk factors were more strongly associated with fatal than non-fatal strokes. Multivariate analyses revealed that, while there was some attenuation of the effect of social class, angina and elevated BP, the risks from atrial fibrillation and diabetes were increased. PMID- 11402348 TI - Implementation and evaluation of local-level priority setting for stroke. AB - We aimed to develop and evaluate a prioritisation process to combine the evidence base with stakeholder involvement within a stroke programme for a Health Improvement Programme (HImP). Implementation involved: formation of a district stroke group (DSG); review of the evidence; survey of DSG members; survey of other key professionals; consensus within the DSG; consultation with local users of the service. Evaluation was through semi-structured interviews and documentary analysis. The process was accepted as appropriate and valuable by the majority of participants, and a district HImP implementation group allocated pound sterling 100 000 for stroke development as a result of this process. However, some felt that stroke itself had been an imposed, rather than an agreed, local priority. The priority setting process was not clear to all participants and change of personnel, particularly in the NHS trusts, led to some perceived lack of ownership. Professionals from secondary care participated, but later criticised the process when they felt that the priorities in the HImP could limit their ability to access money for other service developments. The user consultation days occurred too late to influence the 1999/2002 HImP. We have shown that it is possible to develop an approach that is broadly accepted by stakeholders and balance the evidence base with local ownership. The participation of stakeholders, clarity of procedures, local ownership and awareness of local politics are important in effective priority setting. The model developed will be of value in other settings. PMID- 11402349 TI - Older school children are not necessarily healthier: analysis of medical consultation pattern of school children from a territory-wide School Health Surveillance. AB - It is important to maintain the health of our school children as health is a major factor affecting learning. In developed countries, we assume that school children, especially in higher grades, are healthy because they are relatively free of diseases. However, many of the health problems encountered by school children are not reported in routine health data as they seek help in primary care or by self-medication. In this paper, the medical consultation pattern of school children in Hong Kong was analyzed. It was revealed that school children had a high rate of consultation and self-medication, and the health status of older school children is not better. They were not offered adequate preventive advice. The piecemeal approach to focus on curative medicine will only lead to a high rate of episodic consultation and self-medication without empowering the young people with the skills of self-care and self-help, and positive health behavior. The concept of a health-promoting school with emphasis on more extra curricular health promotion activities, improved communication between health and education sectors, active involvement by pupils and parents, and changing policies and practices would help to make the co-ordinated school health program a greater success. PMID- 11402350 TI - Bicycle-related injuries among the elderly--a new epidemic? AB - Bicycling in Sweden has almost doubled between 1980 and 1992/3 among persons aged 25-64 y. The upward trend is continuing. For the elderly (65 or older) the bicycle is a common means of transport, in both Sweden and a number of other countries. Almost one-third of the Swedish population aged 65 or older bike at some time during the year. The objectives of this study are to describe the pattern of, and trend in, bicycle-related injuries among the elderly in Sweden and to discuss possible means of injury prevention. Mortality data come from official death certificates (1967-96). Hospital-discharge data (1985-96) are also employed, divided into three age groups (-14, 15-64 and 65-), by external cause according to the ICD-9, and also into 'all diagnoses' and 'head injuries'. Number of cases and days of hospital care for persons aged 65 or more, on aggregate and by gender, are reported for 1996. The whole of Sweden and its northern and southern parts separately were investigated. 2830 bicyclists were killed over the period 1967-96, of which 47% were 65 or older. The risk of dying due to bicycling was about 3.7 times greater among the elderly than among children aged 14 or under. There were significant changes in injury trends for all age groups between 1985 and 1996 with regard to hospital care. Annual average decreases for children, of 2.2% for all diagnoses and of 3.4% for head injuries, were observed. For the other two age groups there were increases in all injuries of 3.4% (15-64) and of 2.9% (65-), and increases in head injuries of 4.6% (15-64) and 2.7% (65-). For the elderly living in the southern part of Sweden, there was an increase on average of 2.2% per year over the period, compared with 4.2% for those in the northern part. Males showed a higher incidence of injuries and received longer periods of care than females. Do we have an epidemic of bicycle injuries among the elderly? They face a greater risk of being injured or killed than his/her younger counterpart. For all ages the risk is 7.4 times higher for a bicyclist than for a car driver. The risk for the elderly is about 3 times greater than for the average bicyclist, and as much as 6 times higher for the age group 75-84 y. With some few exceptions, there is no doubt that society has neglected the problem. Decision-makers have a tendency to focus on the relatively young. But people are living longer today and the elderly are healthier, which indicates the need for greater interest and more intervention. We have signs of an epidemic, but one that can be ameliorated or prevented. Just waiting for injury to occur leads only to premature death or lifelong disability. PMID- 11402351 TI - Canadian Aboriginal communities and medical service patterns for the management of injured patients: a basis for surveillance. AB - Growing attention has been placed on injury as a major public health problem which has served to highlight the need for relevant injury data for preventive purposes at the community level. In the case of reserve-based Aboriginal communities in Canada, available injury data, from large datasets, often has little or no relevance at the community level. In addition, the availability of local data is complicated by unique health service and community infrastructures. As such, a prerequisite to establishing injury surveillance requires an understanding of Medical Service Patterns (MSPs) for injured patients intrinsic to a community's health service infrastructure. In determining patterns, cultural and environmental contexts are integral to methodological considerations as historically, Canada's Aboriginal population has been 'controlled' by others in the areas of health, education and social services. The objective of the study was to investigate MSPs in a Canadian Aboriginal community, specific to the management of injured patients, for the purpose of identifying data sites, sources, and collectors. The method relied on a four-step qualitative process designed explicitly for the study community, comprising: (1) semi-structured interviews with key informants; (2) a flow diagram process; (3) focus group discussions; and (4) a summary matrix diagram. This methodology was later replicated with three additional pilot communities. Three major MSPs were identified from nine original patterns generated through the initial data collection process. MSPs were found to be most directly impacted by severity of injury and the proximity of health service providers. Data collection practices were inconsistent, sporadic and poorly coordinated. Data was exclusive to respective data sources and off-reserve documentation was not reported back to the community. MSPs identified key data sites, sources, and collectors relevant to the study population. In conclusion, the four-step qualitative methodology employed in the study was found to be reliable and feasible in identifying community MSPs. Empirical findings confirm the need to investigate MSPs in communities considering surveillance activities, as intra-national differences may be considerable given social inequalities, geographic uniqueness and cultural factors. The use of sophisticated methodologies may detract rather than promote collaborative efforts. PMID- 11402352 TI - Contraceptive use at the family planning clinic of the University of Nigeria Teaching Hospital, Enugu, Nigeria. AB - During the five year review period (January 1993-December 1997), 19,470 clients visited the family planning clinic of the University of Nigeria Teaching Hospital, Enugu. Of these, 2402 clients (12%) were new patients and 17,068 (88%) were old patients. Among the new clients, 2262 (94%) eventually accepted a contraceptive method. The majority of the women (60%) chose the intrauterine contraceptive device (IUCD), 20% chose the injectables, while bilateral tubal ligation and norplant were chosen by 8% and 7%, respectively, of the clients. The oral contraceptive pill was the least popular (1%). Variations in the pattern of contraceptive use among clients at the family planning clinic were discussed. Measures to increase the contraceptive prevalence, and particularly strategies to meet the specific contraceptive needs of clients at the clinic, were also examined. PMID- 11402353 TI - Knowing the risk: relationships between risk behaviour and health knowledge. AB - Communicating risk is a key public health strategy. The implicit assumptions are that the public interprets risk information in a logical fashion and adopts behavioural changes to reduce risk. We assessed risk behaviour, and knowledge and perception of voluntary and involuntary risks using an anonymous questionnaire completed by 472 students. Risk-taking behaviour was measured as the number of different risk behaviours undertaken in the previous 12 months. Knowledge and perception were measured by the extent to which subjects agreed with statements of risk-related information. These varied in complexity from simple statements linking a behaviour with a health risk to numerical statements describing the strength of such relationships. Risk-taking behaviour was highest amongst younger people, males, people whose parents were in non-manual occupations, and people who believed in God (risk-taking behaviour was not related to voting preference or birth order). Overall, knowledge was not significantly related to risk-taking behaviour (P=0.889). However, risk-taking was positively related to more accurate responses to numerical risk questions (P<0.001) and risk-takers were also more likely to perceive both voluntary and involuntary risks as less risky (P<0.05). At least in this cohort, more information about risk is not related to lower risk behaviour. In fact, those individuals with a better understanding of the precise risk associated with certain behaviours were more likely to be higher risk-takers while those who consistently over-estimated risks were low risk-takers. Overall, knowledge and perception of risk explained relatively little of the variance in behaviour. Although these findings need further examination within the general population, public health measures should not assume that information campaigns will necessarily lead to a reduction in risk behaviour. PMID- 11402354 TI - Smoking and alcohol consumption in a UK Chinese population. AB - Little research has been conducted on health in Chinese communities in the UK and there are few representative data on smoking, alcohol consumption or other aspects of lifestyle. We undertook a cross sectional population-based study of 380 Chinese and 625 European men and women aged 25 to 64 y, using self-completion and interview questionnaires in Newcastle upon Tyne, UK between 1991 and 1995. We measured self-reported prevalence of cigarette smoking, number of cigarettes smoked per week and age at starting smoking; self-reported prevalence of alcohol consumption and units of alcohol consumed per week. In age-adjusted comparisons smoking was less common in Chinese (24%) than European men (35%) (P=0.00002) and among Chinese (1%) compared with European women (33%) (P<0.00001). Number of cigarettes smoked was similar among Chinese and European male smokers. Median age at starting smoking was higher among Chinese (18.5 y) compared with European men (15 y) (P=0.00001). Smoking was commonest in older Chinese and in younger Europeans. The prevalence of alcohol consumption was lower among Chinese (63%) than European men (93%) (P<0.00001) and among Chinese (29%) compared to European women (89%) (P<0.00001). Median alcohol consumption was significantly lower among Chinese (2 units/week) than European men (16 units/week) (P<0.00001), and among Chinese (1 unit/week) compared to European women (6 units/week) (P<0.00001). Among those who drank alcohol, Chinese men were less likely to drink above recommended limits than European men (1% vs 39%; P<0.00001). Chinese men and women currently have relatively favourable patterns of smoking compared to European adults in Newcastle. Average alcohol consumption among Chinese who drink is lower than among Europeans, and a substantial proportion of the Chinese population in Newcastle drink no alcohol. Patterns of health related behaviour should be tracked over time in ethnic minority populations to identify changes that pose risk to health and which deserve appropriate intervention. PMID- 11402355 TI - Pros and cons of an illicit drug users' registration system by means of judicial data in Flanders (Belgium). AB - The main objective of this study was to evaluate the advantages and shortcomings of a new kind of registration of illicit drug users in Flanders, Belgium. Data about the profile of drug users who have come into contact with the law were collected by examining the records kept by the District Courts. On the one hand, published data on illicit drug users in Flanders are scarce. On the other hand, a lot of unused judicial information is available. All District Courts in Flanders (n=13) and Brussels (n=1) participated in the study. The data show that a simple registration of verbalised drug users with a short questionnaire can provide useful information for prevention campaigns and police investigation and shows the importance of prevention programmes starting at school. Notwithstanding some shortcomings, the illicit drug users' registration system by means of judicial data in Flanders forms a solid basis upon which an integrated registration of illicit drug use can be built. PMID- 11402356 TI - Characteristics of non-responders to diabetes service use questionnaires. AB - Postal surveys of service use are likely to overestimate health service use and may also underestimate health needs in the population. A historical cohort study, using postal questionnaires and medical records, showed that non-respondents are registered at addresses in more-deprived wards, they are less likely to have attended a hospital diabetes clinic (38% vs 45%) and much less likely to have had a diabetes review in general practice (11% vs 26%). An analysis based on questionnaire respondents would only therefore both underestimate the level of material deprivation and overestimate the proportion receiving routine reviews in general practice in a population with a chronic condition. PMID- 11402357 TI - The cost effectiveness of including pencils and erasers with self-completion epidemiological questionnaires. AB - It is cheap to process epidemiological data from optical mark read (OMR) questionnaires. Respondents should use a pencil to complete OMR questionnaires, but many will not unless these are supplied. Sending pencils and erasers is expensive. Does sending pencils and erasers increase the response rate as cost effectively as sending reminders, or does this decrease the error rate and offset data checking costs? We mailed 300 smokers and half were randomised to receive pencils and erasers. The relative risk (95% confidence intervals) for the response rate for the pencil group relative to the non-pencil group was 0.77 (0.46-1.29) and for the error rate was 1.31 (0.78-2.21). Sending pencils and erasers was not cost-effective in sensitivity analysis with any response rate or using the confidence intervals. Including pencils with mailed epidemiological questionnaires probably has no benefit and any plausible benefit does not offset the costs of sending pencils and erasers. PMID- 11402361 TI - Pain following spinal cord injury. AB - Chronic pain is an important problem following spinal cord injury (SCI) and is a major impediment to effective rehabilitation. The reported prevalence of chronic SCI pain is variable but averages 65% with around one third of these people rating their pain as severe. The mechanisms responsible for the presence of pain are poorly understood. However, evidence from clinical observations and the use of animal models of SCI pain suggests that a number of processes may be important. These include functional and structural plastic changes in the central nervous system following injury, with changes in receptor function and loss of normal inhibition resulting in an increased neuronal excitability. A number of specific types of SCI pain can be distinguished based on descriptors, location and response to treatment. Nociceptive pain can arise from musculoskeletal structures and viscera and neuropathic pain can arise from spinal cord and nerve damage. The role of psychological and environmental factors also needs to be considered. Accurate identification of these pain types will help in selecting appropriate treatment approaches. Current treatments employ a variety of pharmacological, surgical, physical and psychological approaches. However, evidence for many of the treatments in use is still limited. It is hoped that future research will identify effective treatment strategies that accurately target specific mechanisms. PMID- 11402362 TI - Spinal cord blood flow changes following systemic hypothermia and spinal cord compression injury: an experimental study in the rat using Laser-Doppler flowmetry. AB - STUDY DESIGN: It is well known that changes of the body temperature as well as trauma influence the blood flow in the brain and spinal cord. However, there is still a lack of knowledge concerning the levels of blood flow changes, especially during hypothermia. OBJECTIVES: This investigation was carried out to examine the effects of systemic hypothermia and trauma on spinal cord blood flow (SCBF). METHODS: Twenty-four rats were randomized either to thoracic laminectomy only (Th VII-IX) or to 35 g spinal cord compression trauma. The animals were further randomized to either constant normothermia (38 degrees C) or to a systemic cooling procedure, ie reduction of the esophageal temperature from 38 to 30 degrees C. SCBF was recorded 5 mm caudal to the injury zone using Laser-Doppler flowmetry which allows a non-invasive continuous recording of local changes in the blood flow. The autoregulation ability was tested at the end of the experiments by inducing a 30-50 mmHg blood-pressure fall, using blood-withdrawal from the carotid artery. RESULTS: The mean SCBF decreased 2.8% and 3.5% per centigrade reduction of esophageal temperature in the animals sustained to hypothermia with and without trauma, respectively. This could be compared to a decrease of 0.2%/min when only trauma was applied. No significant differences were seen between the groups concerning auto regulatory ability. CONCLUSIONS: Our results indicate that the core temperature has a high impact on the SCBF independent of previous trauma recorded by Laser-Doppler flowmetry. This influence exceeds the response mediated by moderate compression trauma alone. PMID- 11402363 TI - Vestibular caloric stimulation evokes phantom limb illusions in patients with paraplegia. AB - STUDY DESIGN: Prospective study. OBJECTIVES: To determine the mechanisms of body illusions in paraplegia patients as compared with the amputee phantom phenomena. METHODS: A vestibular caloric stimulation was performed in 10 consecutive patients with complete section of the spinal cord. Perception of body, before and after stimulation, was classed as illusion of a normal body (lower limbs with normal morphological, postural and kinetic characteristics perceived as before spinal injury), normal phantom (overly vivid perception of all or part of the lower limbs), deformed phantom (perception of all or part of the limbs below the injury level as abnormal in shape, posture, movement or even number), or painful phantom. RESULTS: After vestibular caloric stimulation, nine out of 10 patients stated their perception of body segments below the injury level had changed to normal phantoms or to deformed phantoms (morphological, postural or kinetic changes). Among the four patients who initially had painful limbs, two stated the stimulation greatly relieved their pain. CONCLUSION: The normal or deformed phantom evoked by vestibular stimulation would result from use of identity data or instantaneous data as is observed in amputees. Cerebral remapping following deafferentation could be the origin of the deformed phantoms. Illusions corresponding to phenomena perceived at the time of the accident corresponding to autobiographical engrammes do not appear to be evoked by vestibular stimulation, as is also the case in amputees. PMID- 11402364 TI - Prophylaxis of thromboembolism in spinal injuries--results of enoxaparin used in 276 patients. AB - OBJECTIVE: To evaluate the results of thromboembolic prophylaxis using enoxaparin in acute spinal injury patients. BACKGROUND: Deep vein thrombosis and pulmonary embolism are major causes of morbidity and mortality in patients with acute spinal injuries. A wide range of thromboprophylactic measures have been proposed. The present study describes the outcome of a regime of enoxaparin and antithromboembolic stockings in acute spinal injuries irrespective of neurological damage. SETTING: Scotland, UK. METHODS: Eighteen-month retrospective review of acute spinal injury patients admitted to a national spinal injuries unit. A thromboembolic prophylactic regimen of early mobilisation, use of antithromboembolic stockings, and subcutaneous administration of enoxaparin 40 mg once a day until patients could be mobilised for more than 4 h per day, was used. Patients with clinical suspicion of deep venous thrombosis or pulmonary embolism were investigated as appropriate. RESULTS: Out of 146 (53% of total) patients with spinal injuries with no neurological deficit only one patient (0.4%) developed clinical evidence of pulmonary embolism and out of 130 (47% of total) with spinal cord injury two (0.7%) developed clinical evidence of deep venous thrombosis while still on enoxaparin. Four patients (1.5%) developed deep venous thrombosis and one (0.4%) pulmonary embolism after discontinuing enoxaparin. There were no fatal pulmonary emboli and one suspected intraspinal bleeding. CONCLUSIONS: The present study suggests that, in addition to physical and mechanical measures, low molecular weight heparin in the form of enoxaparin 40 mg administered once daily compares favourably with previous studies for thromboprophylaxis in acute spinal injuries. PMID- 11402365 TI - Vesico-ureteric reflux in adults with neuropathic bladders treated with Polydimethylsiloxane (Macroplastique). AB - OBJECTIVE: To establish the efficacy of Macroplastique in treating vesico ureteric reflux (VUR) in adults with neuropathic bladder dysfunction. PATIENTS AND METHODS: Fifteen patients (12 male and three female), age range 19 to 80 years (mean age 38) were included in this study. Diagnosis was confirmed by videourodynamics. In seven patients reflux was present bilaterally. Twenty-two refluxing ureters were treated. Twelve patients had detrusor hyper-reflexia, two had areflexic bladders and one had loss of bladder wall compliance. According to the International Grading System, 10 ureters had grade IV reflux, five had grade III reflux, five had grade II reflux, and two had grade I reflux. Macroplastique (0.5-1.5 ml) was injected submucosally under each ureteric orifice to convert the opening to a slit like shape. The patients were followed up from 9 to 68 months. RESULTS: VUR was completely resolved in 72.7% (16) ureters following a single injection and in a further 4.5% (1) ureter following a second injection. 9.1% (2) ureters were improved and treatment failed in 13.7% (3) ureters. Two patients showed a recurrence of reflux 1 and 4 years after primary injection and subsequently had a curative second injection. Most of the patients in whom VUR was cured or improved showed a reduction in laboratory proven urinary infection rates. CONCLUSION: Macroplastique produced an excellent result (86% with complete resolution or improvement of reflux) in treating VUR in adult neuropathic bladders. This is comparable to larger studies carried out on the paediatric population. This is an easy procedure, which avoids major surgery and can be performed as a day case. In cases of failure or recurrence, repeat injection or open surgery can be undertaken without any added complications. PMID- 11402366 TI - The spinal cord independence measure (SCIM): sensitivity to functional changes in subgroups of spinal cord lesion patients. AB - BACKGROUND: The spinal cord independence measure (SCIM) is a newly developed disability scale specific to patients with spinal cord lesions (SCL). Its sensitivity to functional changes in a whole cohort of SCL patients was found to be better than that of the functional independence measure (FIM). OBJECTIVE: o compare the sensitivity to functional changes of the SCIM and the FIM in SCL subgroups. DESIGN: A comparative self-controlled study. SETTING: The Spinal Department, Loewenstein Rehabilitation Hospital, Raanana, Israel. SUBJECTS: 22 SCL inpatients. INTERVENTIONS: Monthly SCIM and FIM assessments of the subgroups. MAIN OUTCOME MEASURES: Functional change detection rate (FDR) and mean differences between consecutive scores (DCS). RESULTS: The outcome measures of the SCIM were higher than those of the FIM for tetraplegia and paraplegia, complete and incomplete lesions (the FIM missed 25-27% of the functional changes detected by the SCIM; DSC 8.2-11.4 vs 5.2-9; P<0.05 in most comparisons). The SCIM did not exhibit this advantage, however, in the functional areas of self care and mobility in the room and toilet. Further subgrouping yielded similar results. CONCLUSIONS: The SCIM is more sensitive than the FIM to functional changes in the subgroups studied, and has the potential to serve as a universal tool for disability assessment of SCL patients. PMID- 11402367 TI - One-stage posterior decompression and fusion using a Luque rod for occipito cervical instability and neural compression. AB - STUDY DESIGN: Original article. OBJECTIVE: The authors present seven cases who underwent one-stage suboccipital, C1 and/or C2 posterior decompression and fusion by Luque rod, wiring and autogenic bone graft for occipito-cervical instability and neural compression. SETTING: Chang Gung University and Medical Center at Kaohsiung, Taiwan. METHOD: Since January 1996, 20 cases of craniovertebral and upper cervical spinal instability were encountered. Seven of these cases had no neurological improvment under Cone-Barton Skeletal traction. Imaging studies revealed poor reduction with persistent neural compression. One patient underwent unsuccessful anterior trans-oral vertebrectomy for decompression. All patients underwent posterior suboccipital craniectomy, C1 and/or C2 laminectomy for decompression. Contoured Luque rod with wiring and autogeneic bone graft was used for craniocervical fusion. After surgical treatment, halo-vest or sterno-occipito mandibular immobilizer (SOMI) was used for 3-6 months. RESULTS: Suboccipital craniectomy and C1 laminectomy could afford a 30-50% increment of anteroposterior diameter in the neural canal and effective decompression of the low medulla and cord. All patients made neurological improvement. The ASIA-MIS scores improved from pre-operative 49 (mean) to 78. Four patients can walk without any support. There were no major complications except for one case requiring readjustment of the halo-vest brace. CONCLUSION: The authors recommend this procedure as one choice for relieving craniocervical instability with neurological compromise. A programmed rehabilitation will afford better neurological improvement. PMID- 11402368 TI - A tribute to the Midlands Centre for Spinal Injuries with the help of princes, pioneers and poets. PMID- 11402369 TI - Consequences of neck manipulation performed by a non-professional. AB - Case report. Documentation of complication of neck manipulation by an untrained person. Tertiary care referral teaching hospital at Lucknow, India. Clinical evaluation, plain radiography of cervical spine, spinal MRI.A 30-year-old man who fainted after neck manipulation by a barber and developed spinal cord and brainstem dysfunction. His MRI revealed an extramedullary, intradural dumbbell shaped mass on the right side at C1 and C2 level compressing the spinal cord. Public awareness should be increased about the danger of neck manipulation by an untrained person especially in the communities where it is commonly practiced. PMID- 11402370 TI - The role of plasma transglutaminase (F XIII) in wound healing of complicated pressure sores after spinal cord injury. AB - STUDY DESIGN: A case report. OBJECTIVES: To demonstrate stimulating action of F XIII in wound healing of complicated pressure sores. SETTING: A Spinal Cord Injury Center in Germany. METHODS: Clinical exam, clinical and photographic wound control, biochemical serum monitoring. RESULTS: Recurrent pressure sores in plegic patients are common complications requiring long-standing conservative or operative therapy. Additional risk factors such as diabetes increase the complication rate for surgery. Surgery itself may be difficult in recurrent pressure sores due to limited remaining soft tissues. We report the case and treatment of a 47-year-old patient with long-standing and recurrent ulcers and complications after flap surgery. As a final option we added plasma transglutaminase (factor XIII) to our treatment scheme which changed the course of the disease dramatically and we achieved complete and rapid healing. CONCLUSION: Our experience suggests that F XIII has a positive role in treating pressure sores as shown already in several other surgical fields. Its use is giving the surgeon an additional tool in complicated cases. PMID- 11402371 TI - An implantable upper extremity neuroprosthesis in a growing child with a C5 spinal cord injury. AB - OBJECTIVES: To implement a functional electrical stimulation (FES) hand neuroprosthesis called the Freehand System in a growing child with spinal cord injury (SCI) using extra lead wire to accommodate limb growth, and to evaluate the performance of the Freehand System during the subject's maturation. SETTING: Pediatric orthopedic hospital specializing in SCI rehabilitation. SUBJECT: Ten year-old female patient with a C5 level SCI. METHOD: The Freehand System was implanted. Eight electrodes were implanted to targeted forearm and hand muscles to provide grasp and release function. The lead wire associated with each electrode was pathed subcutaneously up the arm with 4 cm of extra lead distributed throughout the path to accommodate expected limb growth. All leads were attached to a stimulator placed in the upper chest. Measures of lead unwinding, limb growth, stimulated muscle strength, and hand function were made at 6 and 16 months after implant. RESULTS: By 16 months post implant, the upper limb growth plates were closed and humeral and radial bone growth combined was 2.7 cm from the time of surgery. For all eight leads, lead unwinding in the upper arm was approximately 1.2 cm and was comparable to humeral bone growth (1.4 cm). Lead unwinding in the lower arm was also measurable for the two electrodes in hand muscles. Six of eight electrodes maintained grade 3 or better stimulated muscle strength throughout the growth period according to a manual muscle test. Of the two other electrodes, one appeared to have lost function due to depletion of excess lead. However, hand function with FES was comparable at 6 and 16 months post implant suggesting that growth did not negatively impact performance with the FES system. Hand function with FES was improved over voluntary hand function as well. Using the Freehand System, a pinch force of approximately 15 N was achieved compared to 1.3 N of voluntary tenodesis pinch force. Scores on the Functional Independence Measure (FIM) increased by 9 points when FES was used as compared to voluntary function. Improvements occurred primarily in eating and grooming. Independence in writing was achieved only with FES. CONCLUSIONS: For this child, hand function with the Freehand System was sustained over the growth period and was a significant functional improvement over voluntary hand function. By using excess lead wire, the Freehand System was successfully implemented before skeletal maturity, affording the child improved hand function earlier than would be otherwise indicated. PMID- 11402373 TI - Studies on the expression of mRNA for anion transport related proteins in corneal endothelial cells. AB - PURPOSE: Chloride and bicarbonate are necessary for maintenance of fluid transport by the corneal endothelium, however there is little information on the identity of anion transport proteins that could serve as anion efflux mechanisms in endothelial cells. Therefore, we ask whether mRNA for the anion transport related proteins, CFTR, CLC-2, ClC-3, ClC-5 and AE2, are expressed in human, bovine or rabbit corneal endothelium. METHODS: RT-PCR was performed for CFTR, CLC 2, ClC-3, ClC-5 and AE2 using total RNA from fresh human, bovine and rabbit corneal endothelium as well as cultured bovine corneal endothelial cells (CBCEC). Specificity of PCR products was confirmed by sequencing. RESULTS: RT-PCR analysis gave positive bands at the predicted size for CLC-3 and CLC-5 from fresh human, rabbit and bovine as well as CBCEC. However, for CLC-2, no band was apparent around the predicted size from fresh and cultured corneal endothelium. A band at the predicted size was obtained for CFTR from fresh human, rabbit and bovine endothelium, as well as from CBCEC. RT-PCR analysis for AE2 produced specific bands from fresh human, rabbit and bovine corneal endothelium, but no positive band was obtained from CBCEC. Sequencing analysis further confirmed the identities of CLC-3, CLC-5, CFTR and AE2 in corneal endothelium. CONCLUSIONS: CFTR, CLC-3 and ClC-5 are expressed in fresh and cultured corneal endothelial cells. However, consistent with previous immunoblots studies, AE2 is only expressed in fresh corneal endothelium. These results have implications for modeling possible apical anion efflux mechanisms in corneal endothelium. PMID- 11402374 TI - The conjunctival epithelium in dry eye subtypes: effect of preserved and non preserved topical treatments. AB - PURPOSE: To determine the effect of topical treatments on the conjunctiva in dry eye. METHODS: N = 134 dry eye subjects were diagnosed using a protocol of McMonnies dry eye symptom survey score > 14, fluorescein break up time (FBUT) < 10 s and presence of rose Bengal staining. Differential diagnosis of dry eye subtypes was based on biomicroscopic signs and ocular/medical history. Superficial perilimbal bulbar conjunctival epithelial samples were collected using impression cytology. The nucleo-cytoplasmic ratio (N/C), goblet cell density (GCD) and expression of monoclonal antibodies HLA DR and CD23 were determined. The ocular surface characteristics of untreated subjects, those receiving preserved dry eye treatments and those receiving non-preserved treatments were compared with each other and with controls. Ocular surface characteristics of dry eye subtypes were also examined. RESULTS: An increase in N/C (p = 0.011), reduction in GCD (p = 0.0001) and increase in expression of HLA DR (p = 0.0001) and CD23 (p = 0.0001) were detected in the untreated group compared to controls. No significant differences were found between the group receiving non-preserved dry eye treatments and untreated dry eye group. The group receiving preserved treatments had a reduced GCD (p = 0.0003) and increased expression of HLA DR (p = 0.0003) and CD23 (p = 0.0001) compared to the group receiving non-preserved treatments. Dry eye subtype specific differences in HLA DR and CD23 expression were noted. CONCLUSIONS: The conjunctival inflammation and reduced goblet cell density of dry eye is exacerbated by use of preserved topical agents, and is not significantly improved by use of non-preserved artificial tear supplements alone. Therapeutic strategies for dry eye should aim to increase goblet cell density and control ocular surface inflammation. PMID- 11402375 TI - Human corneal epithelial extracellular matrix perlecan serves as a site for Pseudomonas aeruginosa binding. AB - PURPOSE: Previous data has shown that basement membrane associated perlecan serves as a binding site for Pseudomonas aeruginosa in the wounded mouse cornea. The current study determined whether it also provides a binding site for Pseudomonas aeruginosa in transformed human corneal epithelium. METHODS: Bacterial adherence to transformed human corneal epithelial cells grown in normal or in media containing various inhibitors of glycosaminoglycan synthesis was tested. Bacterial binding was similarly tested in wild-type and in mutant Chinese hamster ovary cell lines naturally deficient in glycosaminoglycan synthesis. Transformed human corneal epithelial extracellular matrix also was tested before and after treatment with anti-proteoglycan monoclonal antibodies or heparinase III before bacterial inoculation. Scanning electron microscopy was used to quantitate adherent bacteria. Intact transformed human corneal epithelial cells or extracellular matrix, the latter either treated or not treated with heparinase III or chondroitin ABC lyase were stained to localize perlecan. RESULTS: Examination of the binding of bacteria to transformed human corneal epithelial cells (normal media vs with inhibitors) and Chinese hamster ovary cell lines suggested that bacterial binding was not associated with the surface of either cell type. In contrast, anti-perlecan antibody, as well as heparinase III decreased the binding of bacteria to corneal extracellular matrix. Fluorescence staining localized perlecan to the extracellular matrix beneath the corneal epithelial cells. CONCLUSIONS: Perlecan localized to the extracellular matrix but not the apical surface of transformed human corneal epithelial cells, provides a binding site for Pseudomonas aeruginosa. PMID- 11402376 TI - Expression of sex steroid hormone receptors in human cornea. AB - PURPOSE: Previously we reported the occurrence of estrogen receptor alpha (ER beta), estrogen receptor beta (ER beta) and androgen receptor (AR) in mouse corneas. The present study was designed to investigate the occurrence of various sex steroid hormone receptors, including ER alpha, progesterone receptor (PR) and AR, in human corneas. METHODS: We used reverse transcription-polymerase chain reaction (RT-PCR) to look for sex hormone receptor mRNAs (ER alpha, PR and AR) in human corneal epithelial cells obtained at autopsy. Next, using an immunocytochemical technique, we localized these receptors in donor human corneas. RESULTS: mRNAs encoding all receptors tested for were found in corneal epithelial cells obtained from male and female donor eyes. Immunocytochemical examination revealed that the receptors were located in the nuclei of corneal epithelial, stromal, and endothelial cells. CONCLUSIONS: Since receptors for both male and female sex hormones are present in human corneas of both genders, we postulate that the receptors may influence the biological functions of corneal cells through direct interaction with specific hormones. PMID- 11402377 TI - Muller cells in the human foveal region. AB - PURPOSE: To investigate the distribution of Muller cells in the foveal region of the human retina. METHODS: After fixation with 4% glutaraldehyde, the percentages of the area of Muller cells were calculated at the macula, posterior pole, equator, and periphery by electron microscopy. After fixation with 4% paraformaldehyde, the silver enhancing technique was applied to show glutamine synthetase (GS) and L-glutamate/L-apartate transporter (GLAST). Furthermore, for the solubilized retinas at each region, Western blot analysis and enzyme-linked immunosorbent assay (ELISA) were performed to detect GS and GLAST in the extracts. RESULTS: The percentages of the area of Muller cells in the outer nuclear layer (ONL) were 12.3 +/- 2.7, 22.1 +/- 4.5, 23.6 +/- 1, and 26.7 +/- 4.5%, respectively. We confirmed less GS and GLAST immunoreactivity in the foveal region. ELISA and Western blot analysis revealed that the amounts of GS and GLAST in the foveal region were smaller than those in any other region. CONCLUSIONS: These results showed that the density of Muller cells is low in the foveal region. PMID- 11402378 TI - Characterization of acute and delayed ocular lesions induced by sulfur mustard in rabbits. AB - PURPOSE: To establish an experimental model for sulfur mustard-induced acute and delayed ocular lesions in rabbits. METHODS: Rabbit eyes were exposed to sulfur mustard (HD) vapor (370, 420 microg/l) for a period of two minutes. A three months follow-up study was carried out, based on the evaluation of clinical, biochemical and histological parameters. RESULTS: HD exposure initiated typical clinical symptoms within 2-6 hrs, characterized by eye closure, eyelid swelling, conjunctival hyperemia, corneal erosions and inflammation. The clinical signs were significantly dose-dependent and reached a peak at 24--72 hrs post exposure. Biochemical evaluation of the aqueous humor exhibited an inflammatory reaction and oxidative stress at 4 hrs after exposure, subsiding at 28 hrs after exposure. Histological examination of corneas at 48 hrs revealed epithelial denudation and marked stromal edema, accompanied by cellular infiltration. Epithelial regeneration started after 72 hrs, and recovery was almost completed within 1--2 weeks, depending on the HD dose. A second phase of pathological processes started as early as two weeks post exposure and was characterized by corneal edema, opacity, recurrent erosions and neovascularization. The delayed injuries were found in 25 and 40% of the eyes respectively, and when appearing, were more severe than the initial ones. CONCLUSIONS: The development of HD-induced ocular lesions in rabbits is similar to the lesions described in human casualties. Quantitative analysis of the various clinical parameters emphasizes the contribution of each tissue to the overall toxic picture. Our experimental model is useful for studying the pathological mechanisms of HD-ocular lesions, and may serve for testing potential therapies. PMID- 11402379 TI - Clinical application of the multifocal VEPs. AB - PURPOSE: To determine whether visual field defects can be detected by the multifocal VEP technique. METHODS: Multifocal VEPs were elicited by a pseudorandom binary m-sequence stimulus (VERIS II). The stimulus was a dartboard like pattern of 61 sectors, and the luminance of each sector alternated between white and black. The stimulus area subtended approximately 25 degrees. Each recording was divided into 8 equal segments, and the total recording time was about 4 min. Multifocal VEPs were recorded from 25 normal subjects and six patients with visual field loss. The responses summed within 4 quadrants were used in the analysis and were compared with the visual fields obtained by perimetry. RESULTS: In six perimetrically-documented visual field defects, the responses summed over each quadrant of the field were reduced in the corresponding affected quadrants. In addition, recovery of the visual field loss following treatment was accompanied by a recovery of the responses. CONCLUSIONS: Multifocal VEPs summed within four quadrants can be used for an objective evaluation of the visual fields. The testing can be obtained in 4 min with no pain or discomfort to the patient. PMID- 11402380 TI - Human beta-defensin 2 is up-regulated during re-epithelialization of the cornea. AB - PURPOSE: Human beta-defensins 1 and 2 (hBD-1, -2) are antimicrobial peptides found in several epithelia including corneal epithelium. Breach of the epithelium leaves the cornea vulnerable to infection so we sought to determine if there is an increase in defensin expression after injury. METHODS: The epithelium from human cadaver corneas was collected by scraping (original samples). The corneas were then placed into organ culture to permit regeneration of the epithelium. Samples of re-grown epithelium were collected when healing was partially and 100% complete as determined by fluorescein staining. Total RNA was extracted from original and re-grown samples and used in RT-PCR reactions using primers specific for hBD-1 and hBD-2 and the constitutively expressed gene glyceraldehyde-3 phosphate dehydrogenase. Immunoblotting was performed to detect defensin peptide in original and re-grown samples. RESULTS: hBD-1 mRNA was detected in all original epithelial tissue samples (n = 10) examined suggesting that it is constitutively expressed. hBD-2 mRNA was detectable in only two of the ten samples. Of six corneas placed in to organ culture, hBD-1 mRNA expression was unchanged in the re-grown epithelial samples compared to the original epithelium samples, however the expression of hBD-2 mRNA was markedly increased. hBD-1 and hBD-2 peptides showed the same pattern of expression as their respective transcripts. CONCLUSIONS: These data show that hBD-2 mRNA and peptide is up regulated in the corneal epithelium during re-epithelialization which is in keeping with the role of this defensin as an antimicrobial peptide. PMID- 11402381 TI - Recovery from hyperemia after overnight wear of low and high transmissibility hydrogel lenses. AB - PURPOSE: To measure the limbal vascular response after 8 hours of eye closure while wearing high and low permeability lenses compared to control eyes without lenses. METHOD: Twenty neophyte participants wore lotrafilcon A silicone hydrogel lenses (HDk; Dk = 140) or etafilcon A hydrogel lenses (LDk; Dk = 18). On two different nights the lenses were randomly worn for 8 hours during sleep in the right eyes only. Left eyes were non-lens wearing controls. Biomicroscopic images of the temporal limbal area were videotaped at baseline, on eye opening and every 20 minutes for 3 hours. A masked observer graded digitized images of the limbal area. RESULTS: On waking and after lens removal there were no differences in hyperemia between the HDk and LDk lens wearing eyes. There were also no differences at any time between the HDk lens wearing eyes and their control eyes (p > 0.05). On waking the eyes wearing the LDk lens were more hyperemic compared to baseline (p < 0.001) and compared to their control eyes at 20 (p < 0.001) and 180 minutes (p = 0.01), indicating slower recovery from hyperemia. The HDk lens wearing eyes recovered to their baseline levels by 180 minutes (p = 0.99), compared to the LDk lens wearing eyes, which had not recovered to baseline levels by 180 minutes (p = 0.04). CONCLUSION: The reduction in hyperemia over time of the HDk lens wearing eyes was the same as the controls. The LDk lens wearing eyes were more hyperemic than the controls on waking and the reduction in hyperemia over time was slower. This suggests that the slower recovery from hyperemia may be affected by the lower oxygen transmissibility of the LDk lens. PMID- 11402382 TI - Quantitative analysis of intravitreal injections in the rat. AB - Intravitreal injections are currently used in the rat to introduce a therapeutic factor in the eye, especially for experimental treatments of retinal degenerations. The injected volume and its location can influence the quantification of results. We have investigated the quantitative effect of a single intravitreal injection in rats at different ages and for different volumes. Albinos rats aged three weeks or two months received intravitreal injections of 1, 3, 5 or 10 microl China ink. Animals were sacrificed immediately after injection, eyes were enucleated, fixated, embedded in paraffin and microtomy was performed in a sagittal plane. Regularly spaced sections were analyzed to reconstruct the vitreous and injected dye volumes. The measured vitreous volume was 6.76 +/- 0.37 mm(3) in three weeks old rats and 13.36 +/- 0.64 mm(3) in two months old rats. Mean intravitreal ink volumes immediately after injection were 0.8 mm(3) for 1 ml injections, 2 mm( 3) for 3 ml, 2.3 to 2.6 mm(3) for 5 microl and 3.2 mm(3 ) for 10 microl. The percentage of vitreous volume involved by the injection ranged from 4.4% to 33.2%. The injected volume is limited by the large lens size of the rat. Extraocular loss of injected solution increases for higher injected volumes, with larger standard deviations. In this model, the dye tends to localize behind the lens. A 3 or 5 microl volume appears to have the best reproducibility with minimum loss of solution. PMID- 11402383 TI - Myopia and contrast sensitivity function. AB - PURPOSE: to measure the contrast sensitivity function of varying degrees of myopia with two types of optical correction (spectacle lens and contact lens). METHODS: One hundred and five myopic eyes and twenty-eight emmetropic eyes were collected. The myopic group included 105 eyes corrected with spectacle lenses and 71 eyes of them corrected with contact lenses, too; all had corrected vision acuity of 20/20 or better. The myopic eyes were divided into four groups: group 1 (-1D to -3D), low myopia; group 2 (-3.25D to -6D), medium myopia; group 3 (-6.25D to -12D), high myopia; and group 4 (> -12D), severe myopia. Spatial contrast sensitivity was measured using the OPTEC 2000 Contrast Sensitivity System. RESULTS: In groups 1 and 2, no statistically significant difference was found between myopes and emmetropes. In group 3, statistically significant loss of contrast sensitivity at higher spatial frequencies was found for myopic subjects corrected with spectacle lens, but not for myopes corrected with contact lens. In group 4, myopic subjects corrected with spectacle lenses showed significantly reduced contrast sensitivity function at all spatial frequencies; subjects corrected with contact lenses showed statistical sensitivity losses at 6, 12, 18 cycle/degree spatial frequencies. CONCLUSIONS: 1. We may suppose that low and medium myopes (groups 1 and 2), who showed normal contrast sensitivity functions, had no retinal dysfunction. 2. For high myopes, contact lens correction could reduce optical defocus and improve contrast sensitivity function in high spatial frequencies. 3. As retinal function disturbances occurred in severe myopes, the diminished contrast sensitivity was not fully compensated by contact lens correction. 4. Loss of contrast sensitivity might be interpreted as evidence for early retinal function disruption before retinal pathological events occur in severe myopes. PMID- 11402384 TI - Isolation of human fetal cones. AB - PURPOSE: To describe a method for isolating a monolayer of human fetal cone photoreceptors and to compare their structure and ultrastructure before and after preparation. METHODS: Eyes from human fetuses (fetal week 20 to 24) were dissected and the neural retina of the developing fovea identified, cut out and placed on 10% gelatin. A VISX Star excimer laser was used to remove the inner retinal layers. The isolated cone monolayers were cultured for 18 hours and compared with untreated retinas by light microscopy and transmission electron microscopy. RESULTS: Excimer laser ablation removed the inner nuclear and ganglion cell layers leaving a monolayer of fetal cones. These cones survive in culture for at least 18 hours. The laser ablation disorganized the ultrastructure of the synaptic pedicles of these cones, left their plasma membranes intact. CONCLUSIONS: The developing central retina of human fetal eyes provides a source of fetal cones, which can be isolated from inner retinal cells using the excimer laser. Such a monolayer of human fetal cones may be useful for transplantation or biochemical studies. PMID- 11402385 TI - Do urinary levels of vascular endothelial growth factor predict proliferative retinopathy? AB - PURPOSE: Vascular endothelial growth factor (VEGF) is elevated in the vitreous of patients with proliferative retinopathies (PR). Angiogenic factors like VEGF are elevated in the urine of subjects with cancers, including those distant from the genitourinary tract. We hypothesized that local increases in VEGF in the vitreous would be reflected in the urine of subjects with PR. METHODS: Urine samples were collected from adults with absent, mild, or severe (requiring laser photocoagulation) PR. VEGF was measured by enzyme linked immunosorbent assay. RESULTS: Of 42 subjects, 16 had no PR and 26 had PR (8 mild, 18 severe). Thirty subjects had diabetes mellitus; 24 of these had PR. Subjects with PR were older than controls. Subjects with PR tended to have higher urinary VEGF (median 123 pg/ml Cr, range 3--1738) than controls without PR (median 93 pg/ml Cr, range 2- 200) (p = 0.08). None of 16 controls, but 11/15 subjects with PR had >200 mg VEGF/mg Cr (p = 0.003), yielding high specificity (100%), but poor sensitivity (42%) of elevated urinary VEGF for PR. Urinary VEGF was also modestly correlated with urinary protein excretion (r(2 ) = 0.23). Correction of VEGF values for urinary protein abrogated any correlation with PR. CONCLUSIONS: Urinary levels of VEGF are associated with PR, but this relationship may be caused by concurrent renal diseases that result in proteinuria and/or renal VEGF production. The insensitivity of the association may preclude its use in screening to avoid eye examinations. PMID- 11402386 TI - The mechanism and change in the optic nerve head (ONH) circulation in rabbits after glucose loading. AB - PURPOSE: To assess the influence of the blood glucose level on ocular capillary circulation, we induced clinically significant hyperglycemia (200--300 mg/dl) in rabbits and investigated the changes in the optic nerve head (ONH) circulation. METHODS: Hyperglycemia was induced by injection of glucose (5.6 mmol/kg) into an auricular vein of healthy albino rabbits and changes in the ONH circulation were measured by the laser speckle method. In order to examine the role of nitric oxide (NO), glucose was administered after intravenous injection of an NO synthetase inhibitor (N(G)-nitro-L-arginine methyl ester, L-NAME, 1 mg/kg), then changes in the ONH circulation were measured. RESULTS: The blood glucose level reached a peak at 30 min after glucose loading and returned to its initial level by 2 hours. ONH circulation showed a 60% increase compared with its initial level at 15 min after glucose loading and subsequently remained almost unchanged throughout the 2-hour observation period. There were no significant changes of the blood pressure, heart rate, or intraocular pressure. The glucose-induced increase of ONH circulation was completely inhibited by pretreatment with L-NAME. CONCLUSIONS: ONH circulation was increased by administration of glucose to healthy rabbits. A high blood glucose level seems to promote ocular capillary circulation and NO as well as insulin appear to have a role in this process. PMID- 11402387 TI - Modeling the vertical motion of a soft contact lens. AB - PURPOSE: The motion of a soft contact lens in the up-down or vertical (inferior superior) and in-out (anterior-posterior) directions drives mixing in the post lens tear film. Thus, it is important to obtain an accurate assessment of lens motion. The commonly used experimental technique to measure the vertical motion, video microscopy, only gauges motion during the interblink period. Since most of the eyelid force, which drives the motion, is exerted during the blink, it is reasonable to assume that the majority of the lens motion in the up-down direction takes place during the blink and is, therefore, hidden from view. Thus, experimentally measured values of vertical lens travel are currently underpredicted. METHODS: In this paper, we use a simple mechanical force balance on the lens to predict its motion in the vertical direction. The forces included in our model are due to the upper and the lower eyelids, gravity, elasticity, and viscous stresses. We input the lens physical parameters, the various tear film thicknesses, and the upper eyelid velocity. Then we integrate a macroscopic force balance to obtain the lens vertical position as a function of time. RESULTS: The proposed model predicts that the downward lens motion during a blink is about 2- 3 times the downward motion during the interblink (centration). CONCLUSIONS: The up-down motion observable during the interblink period is only a small fraction of the total lens vertical travel. PMID- 11402388 TI - Acute effects of H-7 on ciliary epithelium and corneal endothelium in monkey eyes. AB - PURPOSE: Topical or intracameral administration of H-7 doubles outflow facility and reduces intraocular pressure in cynomolgus monkeys, by relaxing and expanding the trabecular meshwork (TM) and Schlemm's canal (SC). Since H-7 may have anti glaucoma potential, we determined its effects on the corneal endothelium and ciliary epithelium for safety considerations. METHODS: Following topical H-7, aqueous humor flow (AHF), corneal endothelial transfer coefficient (k(a)) and anterior chamber (AC) entry of i.v. fluorescein were measured by fluorophotometry; AC aqueous protein concentration ([Protein](AC)) was determined by Lowry assay; and corneal thickness and endothelial cell density and morphology were measured by ultrasonic pachymetry and specular microscopy respectively. Following intracameral H-7, specular and/or light and electron microscopy of the corneal endothelium or ciliary epithelium were performed. RESULTS: Following unilateral topical H-7: (1) AHF and k(a) were essentially unchanged at 0.5--3.0, 3.5--6.0, and 0.5--6.0 hr, with an insignificant increase from 0.5--1.5 hr; (2) [Protein]( AC) was insignificantly increased at 1-1.5 hr but had returned to baseline by 2.5 hr; (3) entry of i.v. fluorescein into aqueous or cornea was modestly and transiently increased; (4) the central cornea thickened significantly at 1--2.5 hr, gradually returning to baseline 2.5 hr after H-7, while peripheral corneal thickness was less affected; (5) corneal endothelial cell borders became indistinct by 1 hr, but cell morphology was recovering by 3- 5 hr and had completely returned to normal by 24 hr; (6) corneal endothelial cell density was unchanged at 5--24 hr. Following intracameral H-7, no significant changes were observed in corneal endothelial cell density or morphology by specular microscopy, nor in corneal endothelial or ciliary epithelial morphology by light and electron microscopy. CONCLUSIONS: A facility-effective intracameral dose of H-7 had no discernible structural effect on the corneal endothelium or ciliary epithelium. It is not yet clear whether carefully chosen topical doses of H-7 or analogues can enhance outflow facility without meaningfully affecting the cornea and ciliary processes. PMID- 11402389 TI - Identification of choroidal ovotransferrin as a potential ocular growth regulator. AB - PURPOSE: In an effort to identify choroidal factors potentially involved in the regulation of ocular growth, proteins released into culture medium of organ cultured choroids were compared between control eyes and eyes recovering from form deprivation myopia. METHODS: The choroids were obtained from the posterior poles of control and recovering chick eyes, and placed into organ culture containing ( 35)S-methionine/(35)S-cysteine. Culture medium was collected after 24 hours and proteins were separated and identified by SDS-polyacrylamide gel electrophoresis (SDS-PAGE), fluorography, immunoprecipitation, western blot analysis and by amino acid sequencing. Choroidal proteins were tested for their effect on scleral proteoglycan synthesis by measuring (35)SO( 4) incorporation into scleral glycosaminoglycans (GAG) in vitro. Choroidal thickness and axial elongation were measured in control and recovering eyes using high frequency A scan ultrasound. RESULTS: The synthesis of an 80 kD protein was greatly increased in the choroids of recovering eyes compared with those of control eyes. Amino acid sequencing and immunoprecipitation indicated that the newly synthesized 80 kD protein was ovotransferrin (transferrin, conalbumin). Ovotransferrin release into the culture medium by isolated recovering choroids was associated with a decrease in the rate of axial elongation in recovering eyes. When tested in vitro, ovotransferrin (500 ng/micro) inhibited scleral proteoglycan synthesis in the sclera by 62% in a dose-dependent manner. CONCLUSIONS: Chick choroids of recovering eyes synthesize and release ovotransferrin during the recovery from form deprivation myopia. Ovotransferrin significantly inhibited proteoglycan synthesis by the sclera, indicating that ovotransferrin may play a role in slowing the rate of vitreous chamber elongation and facilitating the recovery from induced myopia. PMID- 11402390 TI - Postnatal growth retardation exacerbates acidosis-induced retinopathy in the neonatal rat. AB - PURPOSE: We have previously described a metabolic acidosis-induced retinopathy in the neonatal rat, similar to retinopathy of prematurity (ROP). We also have reported exacerbation of oxygen-induced retinopathy by postnatal growth retardation, produced by raising newborn rats in 'expanded' litters. In the present study, we investigated the effect of postnatal growth retardation on the incidence and severity of acidosis-induced retinopathy. METHODS: 100 newborn Sprague-Dawley rats were randomly assigned to two expanded litters of 25 pups each and five standard control litters of 10 pups each. All rats were gavaged with 10 mM/kg NH(4)Cl twice daily from days two to seven. Following five days of recovery, retinal vasculature was assessed using ADPase staining, light microscopy, and computer-assisted image analysis. The presence of neovascularization (NV), severity of NV (clock hours), and vascularized retinal areas, were evaluated in a masked manner. RESULTS: NV occurred in 52% of rats in expanded litters versus 18% of rats in standard control litters (p = 0.005). Postnatal growth retardation of pups in expanded litters was confirmed by comparing total body weight of pups raised in expanded and standard control litters (10.8g vs 13.4g on day 8, p < 0.001; 20.8g vs 25.2g on day 13, p = 0.002). CONCLUSIONS: Postnatal growth retardation increases the incidence of acidosis-induced retinopathy in the neonatal rat. Our study provides further evidence that postnatal growth retardation is a risk factor for preretinal neovascularization in immature retinae and is consistent with the clinical observation that the smallest and sickest premature infants are more likely to suffer from ROP. PMID- 11402391 TI - Intravitreal VEGF and bFGF produce florid retinal neovascularization and hemorrhage in the rabbit. AB - PURPOSE: Vascular endothelial growth factor (VEGF) causes widespread retinal vascular dilation, produces breakdown of the blood-retinal barrier, and is implicated in ocular neovascularization (NV). Basic fibroblast growth factor (bFGF) also has been implicated in the production of ocular NV. This study was performed to investigate the ability of simultaneous sustained intravitreal release of both VEGF and bFGF to induce robust retinal NV in the rabbit. METHODS: Intravitreal implantation of sustained-release Hydron polymeric pellets containing both 20 microg of VEGF and 20 microg of bFGF was performed on adult male Dutch belted rabbits. In other animals either 20 microg or 50 microg bFGF containing pellets was implanted intravitreally; also, either 20 microg VEGF or 50 microg VEGF-containing pellets was implanted. Control rabbits received either blank polymeric pellets or a pellet containing 30 microg bovine serum albumin. Eyes were examined by indirect ophthalmoscopy after surgery at 24 hrs, 48 hrs, 4 days, 7 days, 14 days, 21 days, and 28 days. Findings were documented by color fundus photography and fluorescein angiography (FA). Eyes were enucleated and prepared for histologic analysis at 28 days following intravitreal implantation of the VEGF/bFGF-containing pellets. RESULTS: In all eyes implanted with VEGF/bFGF pellets, dilation and tortuosity of existing blood vessels were observed within 48 hrs after pellet implantation. The progression of retinal vascular changes was rapid and occurred over the entire optic disk and medullary rays between 4 and 7 days. Hemorrhage occurred as early as 14 days after VEGF/bFGF pellet implantation. In eyes with massive hemorrhage, total traction retinal detachment developed after the second week. The presence of abnormal tissues at the vitreo-retinal interface within 28 days was demonstrated by light microscopy while FA showed profuse leakage of dye from anomalous vessels within the first week. Neither bFGF-exposed eyes nor control eyes showed any vascular changes. Eyes that received only VEGF-containing pellets exhibited tortuosity of existing vessels, but neither hemorrhaging nor retinal detachment occurred. CONCLUSIONS: These results demonstrate that retinal vascular changes leading to hemorrhaging is produced rapidly in the rabbit by simultaneous intravitreal release of both VEGF and bFGF. Understanding how these growth factors induce retinal NV may suggest novel therapeutic treatment strategies. PMID- 11402392 TI - Deamidation of Asn-143 of gamma S crystallin from protein aggregates of the human lens. AB - PURPOSE: Determine if gamma S crystallin is preferentially deamidated in the high molecular weight aggregate fraction of old human lenses. METHODS: The high molecular weight aggregate and lower molecular weight fractions were prepared from water soluble proteins of old human lenses. Synthetic peptides corresponding to expected amidated and deamidated tryptic fragments of gamma S crystallin, together with reverse phase chromatography, were used to resolve and quantitate possible deamidation of glutamine-92, glutamine-96 and asparagine-143 from this protein. RESULTS: Analyses of the high molecular weight aggregate from lenses of different ages consistently demonstrated deamidation of asparagine-143, with no deamidation of glutamine-92 or glutamine-96, while analyses of the lower molecular weight fraction from the same lenses showed no detectable deamidation of any of these three residues. CONCLUSIONS: Preferential deamidation of asparagine-143 of gamma S crystallin is present in the high molecular weight fraction of human lenses, consistent with the possibility that modification of this residue may play a role in the aggregation process occurring in vivo. PMID- 11402393 TI - Energy-filtering transmission electron microscopy (EFTEM) in the elemental analysis of pseudoexfoliative material. AB - PURPOSE: To obtain more information on the basic nature of the pathological matrix product accumulating in pseudoexfoliation (PEX) syndrome by analyzing its elemental composition at the subcellular level. METHODS: Energy-filtering transmission electron microscopy (EFTEM), combining the two microanalytical techniques of electron spectroscopic imaging (ESI) and energy-loss spectroscopy (EELS), were performed on ultrathin sections of lens specimens with PEX syndrome using a transmission electron microscope equipped with an integrated electron energy filter. EFTEM is based on inner shell ionization of elements present in the sample giving rise to characteristic signals in well-defined energy-loss regions. The EEL-spectra, demonstrating the presence of a particular element by its specific electron energy-loss edge, were recorded with an integrated scintillator-photomultiplier-system. ESI generated graphic images of elemental localization in the sections after a process of background correction with an IBAS image analysis program. Energy-dispersive X-ray (EDX) analysis of PEX deposits on hydrated lenses was conducted by variable pressure scanning electron microscopy. RESULTS: The ESI element distribution images of both intracapsular and supracapsular PEX material displayed high signals for nitrogen, sulfur, calcium, chlorine, and zinc in clear association with the PEX fibrils. The corresponding EEL-spectra confirmed the data obtained by ESI and showed the presence of the element-specific energy-loss edges. The presence of these elements in PEX fibrils was further confirmed by EDX analysis. No specific signals were obtained for phosphorus, oxygen, or aluminum. CONCLUSIONS: This study demonstrates the presence of nitrogen, sulfur, chlorine, zinc, and calcium both in mature and in aggregating PEX fibrils of the lens capsule. EFTEM proved to be a highly sensitive method for the microanalytical study of biological material with unknown composition, such as PEX material, at the subcellular level. PMID- 11402394 TI - Verbal and affective laterality effects in P-dyslexic, L-dyslexic and normal children. AB - Lateralization of verbal and affective processes was investigated in P-dyslexic, L-dyslexic and normal children with the aid of a dichotic listening task. The children were asked to detect either the presence of a specific target word or of words spoken in a specific emotional tone of voice. The number of correct responses and reaction time were recorded. For monitoring words, an overall right ear advantage was obtained. However, further tests showed no significant ear advantage for P-types, and a right ear advantage for L-types and controls. For emotions, an overall left ear advantage was obtained that was less robust than the word-effect. The results of the word task are in support of previous findings concerning differences between P- and L-dyslexics in verbal processing according to the balance model of dyslexia. However, dyslexic children do not differ from controls on processing of emotional prosody although certain task variables may have affected this result. PMID- 11402395 TI - Response inhibition and measures of psychopathology: a dimensional analysis. AB - On the basis of Quay's (1988a, 1988b, 1993, 1997) model in which the Behavioral Inhibition System (BIS) and the Behavioral Activation System (BAS) are linked to various forms of child psychopathology, predictions were made regarding the relation between inhibitory control and two dimensions of psychopathology: externalizing and internalizing behavior. Inhibitory control was measured using two versions of Logan and Cowan's stop signal paradigm (1984; Logan, Cowan, & Davis, 1984; Osman, Kornblum, & Meyer, 1986, 1990). The primary outcome measure for the stop tasks was stop signal reaction time (SSRT) which measures the latency of the inhibition process. A positive relationship was predicted for externalizing behavior, whereas a negative relationship was predicted for internalizing behavior. A total of 42 non-clinical elementary school children, in the age range of 6 to 12 years, participated in the study. Externalizing behavior was positively related to response inhibition. Symptoms of ADHD seem to be better at predicting inhibitory functioning than symptoms of aggressive behavior disorders. Some support was found for a negative relation between internalizing behavior and inhibitory control. These findings support Quay's model and the discriminant validity of inhibitory control with regard to externalizing and internalizing behavior. PMID- 11402396 TI - The association of neurofibromatosis type 1 and attention deficit hyperactivity disorder. AB - Some research and clinical observations have linked Neurofibromatosis Type 1 (NF 1) and Attention Deficit Hyperactivity Disorder (ADHD). In order to investigate whether ADHD is part of the phenotype of NF-1 or is a separate, unrelated disorder within families, we compared the ADHD status of children affected with NF-1 to that of their unaffected-NF-1 siblings and to that of their biological parents. Results of matched-pair analyses were calculated and revealed a significant with-in pair discordance, when comparing children with NF-1 and their siblings and when comparing children with NF-1 and their biological parents (in families with a sporadic, non-familial NF-1 child). These findings suggest that ADHD may occur as a component of the NF-1 phenotype. PMID- 11402397 TI - Prevalence and correlates of depressive symptoms following traumatic brain injuries in children. AB - The prevalence and correlates of depressive symptoms following childhood traumatic brain injuries (TBI) were examined using data drawn from a prospective longitudinal study. Participants included 38 children with severe TBI, 51 with moderate TBI, and 55 with orthopedic injuries (OI). Assessments occurred shortly after injury (baseline) and at 6- and 12-month follow-ups. Children completed the Child Depression Inventory (CDI). Parents rated depressive symptoms using the Child Behavior Checklist (CBC), with baseline ratings reflecting premorbid status. Assessments also included measures of children's neurocognitive functioning and the family environment. The three groups did not differ overall in self-reported symptoms on the CDI, but did display different trends over time. The three groups did not differ on parent ratings of premorbid depressive symptoms on the CBC, but parents reported more depressive symptoms in the TBI groups than in the OI group at 6- and 12-month follow-ups. Child and parent reports were correlated for children in the TBI groups, but not for those in the OI group. Depressive symptoms were related to socioeconomic status in all groups. Socioeconomic status also was a significant moderator of group differences, such that the effects of TBI were exacerbated in children from more disadvantaged homes. Although self-reports of depressive symptoms were related inconsistently to children's verbal memory, parent reports of depressive symptoms were unrelated to IQ or verbal memory. The findings suggest that TBI increases the risk of depressive symptoms, especially among more socially disadvantaged children, and that depressive symptoms are not strongly related to post-injury neurocognitive deficits. PMID- 11402398 TI - A clinical comparative study evaluating the effect of epilepsy versus ADHD on timed cognitive tasks in children. AB - Both children with epilepsy and children with ADHD may be characterized by slowing on reaction-time measurement. This is of particular interest, as neuropsychological assessment is often requested in the differential diagnosis between children with short non-convulsive epileptic seizures and children with ADHD. In this study we attempt to identify patterns of impairment on timed tasks that are specific for epilepsy, relative to ADHD. This study was an open, controlled parallel-group clinical investigation which included two groups of patients: 60 children with ADHD and 60 children with epilepsy. These children were compared with a control group (n=30) on two types of timed cognitive tasks: tasks with low information load (simple reaction-time measurement) and tasks with high information load (multiple decision reaction-time measurement). The simple reaction-time measurements show significant differences between ADHD and controls (all except for visual RT non-dominant hand) and between epilepsy and controls (only one test). No significant differences were found between epilepsy and ADHD. The two tests with high information load show significant slowing compared with the controls for epilepsy on the Binary Choice Reaction-Time Test and for ADHD on the Visual Searching Test. On both tests also the differences between epilepsy and ADHD are significant. The two tests in combination have a relatively satisfactory potential to classify the children with ADHD (75% correct classification) and the children with epilepsy (55% correct classification). We may conclude that complex reaction-time tests (i.e., timed tasks with high information load) have potential for assessing the differential impact of ADHD and epilepsy on attentional function. These tasks specifically reveal general slowing for children with epilepsy and slowing as an effect of failures of inhibitory self control on unstructured tasks for ADHD. PMID- 11402399 TI - Processing speed in children with attention deficit/hyperactivity disorder, inattentive type. AB - Attention Deficit/Hyperactivity Disorder (ADHD) is among the most common and most often reconceptualized neurobehavioral disorders of childhood. In the most recent DSM-IV, a primarily inattentive subtype of ADHD (AD) has again been identified. This study explores the neuropsychological profile of this group of children. Eighty-two children referred for school-related problems participated. Twenty five met criteria for AD; 52 met criteria for reading disability (RD); 9 were comorbid for RD and AD. AD children performed poorly on measures of information processing speed. Children with comorbid AD/RD were distinguishable from those with RD on speed of processing measures only. Vulnerability to information processing load may be at the root of many of the behavioral manifestations of AD. PMID- 11402400 TI - A common ancestral origin of the frequent and widespread 2299delG USH2A mutation. AB - Usher syndrome type IIa is an autosomal recessive disorder characterized by mild to-severe hearing loss and progressive visual loss due to retinitis pigmentosa. The mutation that most commonly causes Usher syndrome type IIa is a 1-bp deletion, described as "2299delG," in the USH2A gene. The mutation has been identified in several patients from northern and southern Europe and from North America, and it has been found in single patients from South America, South Africa, and China. Various studies have reported a range of frequencies (.16-.44) among patients with Usher syndrome, depending on the geographic origin of the patients. The 2299delG mutation may be the one that most frequently causes retinitis pigmentosa in humans. Given the high frequencies and the wide geographic distribution of the mutation, it was of interest to determine whether the mutation resulted from an ancestral mutational event or represented a mutational hotspot in the USH2A gene. Haplotype analysis was performed on DNA samples from 116 unrelated patients with Usher syndrome type IIa; the patients were from 14 countries and represented 148 2299delG alleles. On the basis of six single-nucleotide polymorphisms within the USH2A gene, 12 core haplotypes were observed in a panel of normal chromosomes. However, in our analysis, only one core haplotype was found to be associated with the 2299delG mutation. The data indicate that the widespread geographic distribution of the 2299delG mutation is the result of an ancestral mutation that has spread throughout Europe and into the New World as a result of migration. PMID- 11402401 TI - Posterior cruciate-sacrificing versus posterior cruciate-substituting total knee arthroplasty: a study of clinical and functional outcomes in matched patients. AB - Since the introduction of the Total Condylar Prosthesis (TCP) in 1974, concomitant improvements in surgical technique and prosthetic design have occurred. The individual effects of each of these variables have not been investigated, however. This study evaluates 2 different knee designs using the same operative technique by a single surgeon. All primary total knee arthroplasties performed between 1986 and 1989 were entered into a database of 2 cohorts, TCP and Press Fit Condylar (PFC) knees, matched for age, gender, body weight, and diagnosis. Follow-up data within 12 months of each other were used, evaluating patients clinically and using a self-administered questionnaire. In each cohort, 74 knees were matched by these criteria. Follow-up time was 4.04 and 4.45 years for the TCP and PFC cohorts. Range of motion averaged 107 degrees and 112 degrees for the TCP and PFC cohorts. This difference was statistically significant. Total knee score and functional score improved significantly. Anterior knee pain was present in 9 TCP and 3 PFC knees. Lateral release was performed in 30 TCP and 18 PFC knees. The PFC showed an advantage in ROM, stair function, anterior knee pain, and use of lateral release. Both designs showed comparable pain relief and walking ability. PMID- 11402402 TI - Cementless femoral fixation in the rheumatoid patient undergoing total hip arthroplasty: minimum 5-year results. AB - Sixty-two total hip arthroplasties in 49 patients with a diagnosis of rheumatoid arthritis were performed between November 1986 and December 1992. All components were titanium alloy with a circumferential plasma-spray porous coating. Four patients (4 hips) died before 5-year follow-up, and 6 patients (8 hips) were lost to follow-up, leaving 39 patients (50 hips) for review at a minimum 5-year follow up after surgery (mean, 8 years; range, 5-12 years). There were 12 men and 27 women, with a mean age at time of surgery of 55 years (range, 25-77 years) and a mean weight of 69 kg (range, 42-109 kg). Compared with the preoperative Charnley scores, there was significant improvement in the postoperative scores: pain, from 2.7 to 5.7, and function, from 3.2 to 5.3. Thigh pain was present in 1 patient (1 hip) (2.0%). No femoral fractures occurred intraoperatively with the insertion of the prosthesis. Spot welds consistent with bone ingrowth were identified in all of the femoral components. No femoral components showed evidence of radiographic loosening or required revision for aseptic loosening or incapacitating thigh pain, but 7 acetabular revisions were performed. Uncemented femoral fixation with this component design in rheumatoid patients appears to be a promising treatment. PMID- 11402403 TI - Instability after total knee arthroplasty with the Miller-Gallante II total knee: 5- to 7-year follow-up. AB - A consecutive series of 64 posterior cruciate-retaining hybrid total knee arthroplasties in 50 patients were reviewed with an average 57-month follow-up. Seven patients died or were lost to follow-up. One patient was revised for infection in the early postoperative period, and there were no cases of aseptic loosening. Four revisions were performed for recurrent effusions and progressive instability at a mean of 55 months postoperatively. There were statistically significant correlations between manual stability testing (anteroposterior, mediolateral, and pivot shift) and Hospital for Special Surgery and Knee Society scores. At intermediate follow-up, the Miller-Gallante II total knee arthroplasty is functioning well except in a subgroup of patients with progressive instability resulting from a combination of patient and implant factors. PMID- 11402404 TI - Management of knee pain and stiffness after total knee arthroplasty. AB - This article reviews 33 patients who presented with persistent pain, inadequate knee motion, or both after total knee arthroplasty. Of 33 patients, 26 had inadequate motion treated by closed manipulation, arthroscopic manipulation, or a modified open release manipulation. In 23 patients, these procedures were successful. Of the 26 patients, 85% had a history of previous knee surgery or diabetes mellitus. The average gain in range of motion was not different in comparing early (<12 weeks) manipulation versus late (>12 weeks) manipulation. Seven of the 33 patients presenting with pain, swelling, inadequate motion, or snapping sensations had either failure of bonding between the polymethyl methacrylate and the components (4 patients) or painful fibrous intra-articular bands (3 patients). These patients were treated successfully either by recementing the components or by arthroscopic fibrous band release. PMID- 11402405 TI - Continuous femoral blocks improve recovery and outcome of patients undergoing total knee arthroplasty. AB - This study was designed to determine the effects of continuous femoral infusion (CFI) on total knee arthroplasty recovery. A total of 92 patients were distributed in 3 groups: Patients in group 1 received general anesthesia followed by patient-controlled analgesia (PCA) with morphine (n = 33), patients in group 2 received 3-in-1 and sciatic blocks followed by CFI (n = 29), and patients in group 3 received epidural analgesia (n = 30). Blocks reduced postoperative morphine requirement by 74% (vs group 1; P<.05) and 35% (vs group 3; P<.05). Blocks provided better recovery than PCA with morphine or an epidural. The use of CFI was associated with a reduction of postoperative bleeding by 72% (vs group 1; P<.05) and allowed better performance on continuous passive motion. CFI was associated with a 90% decrease in serious complications and a 20% decrease in the length of hospitalization. CFI represents a better alternative than PCA or epidural analgesia for postoperative pain management and immediate rehabilitation after total knee arthroplasty. PMID- 11402406 TI - Activity level and wear in total knee arthroplasty: a study of autopsy retrieved specimens. AB - We assessed the correlation between activity level, length of implantation (LOI), and wear in total knee arthroplasty. Twenty-eight implants were retrieved at autopsy from 8 men and 15 women. Linear, volumetric, and visual wear and the presence or absence of creep were quantitated. Functional level was classified using the Knee Society, the standard Charnley classification, and the UCLA activity level scale. The average age at surgery was 68 years +/- 14.0 SD and average LOI was 74 months +/- 38 SD. The average linear and volumetric wear rates were 0.127 mm/y +/- 0.104 SD and 31.80 mm3/y +/- 42.8 SD. LOI (B coefficient = 0.656 +/- 0.0 SE; P<.001) correlated with linear, volumetric, and visual wear rates. Charnley C patients showed decreased volumetric wear in the lateral compartment (P=.01). Decreased activity level (UCLA) correlated with areas of less extent and severity of creep (P=.001 and P<.001). PMID- 11402407 TI - Subvastus approach for total knee arthroplasty: a prospective, randomized, and observer-blinded trial. AB - A prospective, randomized, and blinded trial was conducted with 89 consecutive primary knee arthroplasties comparing standard medial parapatellar arthrotomy with the subvastus approach. All patients received the same prosthesis (Insall Burstein II) inserted by one surgeon using an identical technique with the only difference being the approach. The parapatellar approach (group I) was used in 43 knees, and in the remaining 46 knees the subvastus approach (group II) was used. Assessment revealed significantly earlier return of straight-leg raise (3.2 days vs 5.8 days, P <.001), lower consumption of opiates in the first week (78 mg vs 102 mg, P <.001), less blood loss (527 mL vs 748 mL, P <.001), and greater knee flexion at 1 week (78 degrees vs 55 degrees, P <.001) in group II (subvastus approach). The subvastus approach offers early advantages over the standard parapatellar arthrotomy. It preserves the integrity of the vastus medialis and peripatellar plexus of vessels. We advise its wider use in primary total knee arthroplasty. PMID- 11402408 TI - Comparison between 1-hour and 24-hour drain clamping using diluted epinephrine solution after total knee arthroplasty. AB - A prospective study was conducted to determine the optimal clamping time in the drain-clamping method after total knee arthroplasty. In a randomized trial, 44 primary total knee arthroplasties were studied after the drain-clamping method using diluted epinephrine solution was applied for 1 hour or 24 hours. The mean blood loss into the drains was less in the 24-hour group compared with the 1-hour group (35 mL in the 24-hour group and 247 mL in the 1-hour group). No statistically significant difference was found in hemoglobin or hematocrit levels between the 2 groups after surgery. Only 1 patient (4.5%) in the 1-hour group and 2 patients (9.1%) in the 24-hour group received blood transfusion. There were significantly more complications in the 24-hour group (P <.05). Two patients (9.1%) in this group required additional surgery. One-hour clamping is preferable to 24-hour clamping in the drain-clamping method for minimizing complications. PMID- 11402409 TI - Head penetration into Hylamer acetabular liner sterilized by gamma irradiation in air and in a nitrogen atmosphere. AB - We reviewed 25 consecutive primary cementless total hip arthroplasties with Hylamer acetabular liners (Hylamer group) and 12 with conventional ultra-high molecular weight polyethylene (Enduron group). Two-dimensional penetration of the femoral head into the liner was determined from anteroposterior radiographs of the pelvis. Head penetration rate was 0.37 mm/y in the Hylamer group sterilized by gamma irradiation in air (n = 6; mean length of follow-up, 3 years), 0.21 mm/y in the Hylamer group sterilized by gamma irradiation in a nitrogen atmosphere (n = 19; mean length of follow-up, 2.7 years), and 0.11 mm/y in the Enduron group (n = 12; mean length of follow-up, 3.9 years). Osteolysis was identified in 6 of the 25 hips with Hylamer liners and 1 of the 12 hips with conventional liners. There was a positive linear correlation between period from production to operation and head penetration rate with Hylamer liner sterilized by gamma irradiation in air and no correlation in a nitrogen atmosphere. Rapid oxidation by irradiation in air might not be the main cause of high rate of wear in Hylamer liners. PMID- 11402410 TI - Primary total knee arthroplasty for supracondylar/condylar femoral fracture in osteoarthritic knees. AB - Supracondylar or condylar femoral fractures require extended treatment and often result in loss of knee range of motion. We present 3 cases in which femoral fractures and preexisting painful knees secondary to osteoarthritis were treated using total knee arthroplasty with a stemmed femoral implant. The patients were all women aged 83, 84, and 87 years old. All fractures united within 3 months, and the patients could walk with 1 cane within 2 months. Primary total knee arthroplasty should be considered as a treatment for supracondylar/condylar femoral fractures in osteoarthritic knees. PMID- 11402411 TI - Patient satisfaction compared with general health and disease-specific questionnaires in knee arthroplasty patients. AB - When assessing the health status of patients after orthopaedic surgery, such as knee arthroplasty, general health and disease-specific questionnaires are gaining in popularity because of their precision in detecting subtle differences. Self administered postal surveys using extensive questionnaires have associated patient burden, however, which may have an impact on response rate and completeness. When a high response rate is important or when the use of comprehensive questionnaires is not practical, it may be possible to gain useful outcome data after a surgical procedure by simpler means. Two postal surveys to knee arthroplasty patients were performed. In the first survey, we posed a simple question regarding patient satisfaction to 27,114 patients. A second survey was sent 9 months later to 3,600 of the same patients; the same simple satisfaction question was posed along with several previously validated general health (NHP, SF36, SF12) and disease/site-specific (Oxford-12, WOMAC) outcome questionnaires. We found that patient satisfaction correlates significantly with general health and disease-specific outcome measures, with the highest correlation to the domains that relate to pain and function. When sent a simple satisfaction questionnaire, 95% of the patients answered, whereas the usable return rate of the more comprehensive questionnaires was 18% to 45% lower. Patients not responding to the comprehensive questionnaires were more often unsatisfied with their operated knee than patients responding. PMID- 11402412 TI - Rehabilitation after total shoulder arthroplasty. AB - Physical therapy constitutes an essential determinant of clinical outcome after total shoulder arthroplasty. We reviewed our results in 81 shoulders at a minimum of 2 years' follow-up, with specific focus on the maintenance of motion and the development of soft tissue healing problems. Our findings show that our graduated rehabilitation program allows most patients to obtain motion comparable to that possible intraoperatively with few complications. Of patients, 70% maintained their elevation, and 90% maintained external rotation. Patients with a diagnosis of rheumatoid arthritis, traumatic arthritis, and osteonecrosis were identified as being at risk for failure to regain motion and for tendon healing complications. PMID- 11402413 TI - The effect of a tibial baseplate undersurface peripheral lip on cement penetration in total knee arthroplasty. AB - Many total knee arthroplasty (TKA) tibial baseplates have a peripheral lip or cement pocket theoretically to increase cement penetration by decreasing edge escape of the cement squeeze film on baseplate impaction. We reviewed 177 consecutive TKAs (89 nonlipped and 87 lipped baseplates) performed by a single surgeon, using the same third-generation tibial cementation techniques. Cement penetration patterns of the lateral 12 mm of proximal tibia were examined by a semiautomatic computerized digital image analysis technique. Penetration in the lipped baseplate was double that of the nonlipped baseplate at the component's lateral edge (P<.05) and continued to be statistically significantly greater to 4 mm from edge. From 5 to 7 mm, the difference was no longer statistically significant, and from 8 to 12 mm, the penetration was equal. PMID- 11402414 TI - Computed tomography measurement of the surgical and clinical transepicondylar axis of the distal femur in osteoarthritic knees. AB - To assess rotational alignment of the distal femur, 2 types of transepicondylar axes have been used in the literature. We compared surgical and clinical epicondylar axes in the measurement of rotational alignment of the femur in planning for total knee arthroplasty (TKA). We examined 48 patients who were candidates for TKA. Computed tomography images of both knees were obtained, and condylar twist angle and posterior condylar angle were measured. The medial sulcus was detected in only 33 knees. The more severe the grade of osteoarthritis, the more difficult it was to detect the medial sulcus. The most prominent point of medial epicondyle was detectable in all knees. We recommend the use of the clinical epicondylar axis in computed tomography measurement in selective planning for TKA. PMID- 11402415 TI - Aspergillus fumigatus infection in a mega prosthetic total knee arthroplasty: salvage by staged reimplantation with 5-year follow-up. AB - Fungal infection after total joint arthroplasty is an extremely serious complication and a challenge to the treating physician. When a fungal infection is compounded by a massive allograft or a metallic segmental replacement of the femur or other long bone, treatment options become increasingly limited and commonly have led to arthrodesis or amputation of the infected limb. We present the first case report of a low-grade osteosarcoma treated with a segmental distal femoral allograft prosthetic composite knee arthroplasty, which was complicated by infection with Aspergillus fumigatus. This complication was treated successfully with a staged reimplantation procedure, intravenous amphotericin, and oral fluconazole suppression. At 5 years after reimplantation, the patient has had no evidence of infection, no pain, and excellent range of motion without extensor lag. The Knee Society knee score improved from 50 to 100, and the function score improved from 40 to 100 (for both scores, 100 is the maximum). PMID- 11402416 TI - Closed reduction of a dislocation of a constrained acetabular component. AB - A constrained acetabular liner occasionally is used in management of chronic instability after total hip arthroplasty. If dislocation occurs out of a constrained liner, open reduction is indicated. A case is presented of closed reduction of a femoral component into a constrained liner. PMID- 11402417 TI - Simultaneous management of ipsilateral gonarthritis and ununited tibial stress fracture: combined total knee arthroplasty and internal fixation. AB - Nonunion of a proximal tibial stress fracture is rare and can be difficult to manage, especially if associated with ipsilateral gonarthritis. Three patients with nonunion of a proximal tibial stress fracture adjacent to an arthritic knee joint were managed by performing simultaneous total knee arthroplasty and internal fixation of the fracture site. The technique involved performing a total knee arthroplasty with angular correction at the site of the extra-articular and intra-articular deformity, bone grafting of the nonunion site, and stabilization of the fracture with a long uncemented intramedullary stemmed tibial component and a unicortical plate. Fibular ostectomy was required during the index surgery in 2 cases to achieve the desired angular correction. In all 3 patients, healing of the nonunion site and limb realignment was achieved. There were no complications or infections associated with the surgery. All patients progressed to full weight bearing at 3 months and had clinical and radiographic union of the nonunion site at 6 months. PMID- 11402418 TI - Rare case of sciatic nerve palsy in a modular total hip arthroplasty. AB - A rare case of sciatic nerve palsy in an uncemented modular hip arthroplasty is described. We believe that a delay in recognizing liner dissociation causing granuloma was responsible for the nerve palsy. Delay in diagnosis can be avoided by a careful inspection of prereduction and postreduction radiographs. PMID- 11402419 TI - Surgical treatment of obturator nerve palsy resulting from extrapelvic extrusion of cement during total hip arthroplasty. AB - A female patient was successfully treated surgically for obturator nerve palsy resulting from extrapelvic extrusion of cement during total hip arthroplasty. Obturator nerve palsy developed because of the extrapelvic extrusion of cement medially in the anterior part of the incisura acetabuli in the zone of the extrapelvic exit of the obturator tunnel. Compression of the obturator nerve was confirmed by electromyography. After surgical removal of a piece of cement, 2.5 cm in length, which was done 11 months after total hip arthroplasty, and after 6 months of intensive postoperative rehabilitation, the patient was able to return to everyday activities and normal working habits. After 1 year, clinical and electromyography results were almost completely normal. A review of current knowledge regarding obturator nerve injuries resulting from total hip arthroplasty is presented. PMID- 11402420 TI - Parsonage-Turner syndrome after total-hip arthroplasty. AB - A 68-year-old patient developed Parsonage-Turner syndrome after total hip arthroplasty. There was an acute onset of intense pain in the shoulder 48 hours after surgery followed by complete paralysis of the shoulder and almost complete loss of strength in the arm. Recovery was slow, and mild weakness persisted 7 years after the operation. Parsonage-Turner syndrome has been associated with various surgical procedures but has not been reported after total hip arthroplasty. For medicolegal reasons, it should be distinguished from a traction injury of the brachial plexus or compression of the radial or ulnar nerve resulting from positioning of the patient. Parsonage-Turner syndrome should be considered in the differential diagnosis when a patient describes pain or weakness in the shoulder after joint arthroplasty. PMID- 11402421 TI - Intravenous polymethyl methacrylate after cemented hemiarthroplasty of the hip. AB - Current cementing techniques used during hip arthroplasty aim to maximize the bond at the bone-cement interface in an effort to increase the longevity of the prosthesis. To accomplish this, one must generate high intramedullary pressures, which are known to be associated with complications such as cement implantation syndrome. We record a rare complication following cement pressurization of a hip hemiarthroplasty that resulted in intravenous polymethyl methacrylate (PMMA). This complication however, is not associated with a significant morbidity or mortality, but it is important to identify and distinguish from a femoral cortical defect, which can be created during surgery. PMID- 11402422 TI - A new technique of limb salvage after infected revision total knee arthroplasty: artificial fusion. AB - Limb salvage after infection and multiple knee revision arthroplasty is difficult. The treatment options and a brief history of artificial fusion are reviewed, and a case with 4-year follow-up is described. PMID- 11402423 TI - Staged reimplantation of total knee arthroplasty after Candida infection. AB - Prosthetic joint infection with Candida is uncommon. Only 28 cases have been reported in the English literature. Successful reimplantation after eradication of Candida infection has been reported in 3 hip joints and only 1 knee. We present the case of a 68-year-old woman with chronic Candida parapsilosis infection of a prosthetic knee joint. Removal of the prosthesis, thorough debridement, and antifungal therapy treated the infection successfully. Antifungal therapy included 6 weeks of parenteral administration of fluconazole followed by 4 weeks of oral fluconazole. The involved knee joint was reimplanted 3 months after initial treatment. The prosthetic joint was pain free and functioned satisfactorily during the ensuing 4 years. No recurrence of infection was noted. The principle in treating Candida prosthetic infection generally has been the same as that of bacterial prosthetic infection. In chronic cases, removal of implants, thorough debridement, and effective antifungal therapy are mandatory for the eradication of infection. Reimplantation of the prosthesis can be performed successfully in a staged surgical procedure with the interval between the 2 stages shortened to 3 months. PMID- 11402424 TI - Revision knee arthroplasty using impaction grafting and primary implants. AB - A technique is described for revision knee arthroplasty with impaction grafting of cancellous bone grafts to treat bone loss and the use of primary implants. Specially designed impaction instruments were used. Bone loss was classified according to the Anderson Orthopaedic Research Institute classification. Four year follow-up of 11 knees in 9 patients showed good clinical and radiographic results. PMID- 11402425 TI - A simple technique to remove well-fixed, all-polyethylene cemented acetabular component in revision hip arthroplasty. AB - Well-fixed, all-polyethylene acetabular components may have to be removed in cases of recurrent dislocations, infection, or fracture of ceramic femoral heads. We describe a simple technique using acetabular reamers that allow cup thinning. Through this technique, the polyethylene rigidity is diminished considerably, allowing its easy removal without any risk of fracture, particularly of the acetabulum walls. PMID- 11402426 TI - The effect of polymethyl methacrylate on latex-free surgical gloves. AB - In the current medical environment of increased awareness about latex sensitivity, more patients are presenting with known latex allergy. One such patient presented to us for total knee arthroplasty. During this procedure, we noticed holes in our latex-free surgical gloves when polymethyl methacrylate (PMMA) was being handled. Through subsequent testing of different brands of latex free surgical gloves with various brands of bone-cement, we found that Allergard latex-free surgical gloves are destroyed by contact with PMMA. Biogel Neotech and Duraprene latex-free gloves show no such destruction. These results indicate that Allergard gloves should not be used in procedures in which bone-cement is employed. Biogel Neotech and Duraprene latex-free gloves are an acceptable alternative. PMID- 11402427 TI - The use of the cystoscope in revision hip arthroplasty. PMID- 11402428 TI - Accuracy of soft tissue balancing in total knee arthroplasty. PMID- 11402430 TI - Breast cancer at the millennium: the new frontier. PMID- 11402431 TI - Breast cancer imaging: the future. AB - Imaging technology has changed rapidly over the last four decades. This is especially true in the area of breast cancer. While anatomic delineation of breast tumors was considered a crowning achievement of the late 20th century, most recently the major focus has shifted toward physiologic and even molecular level tumor detection. The goals of imaging are threefold: (1) the earliest possible detection of primary breast tumors, (2) correlation of imaging results with other clinical parameters to assess disease biology, and (3) accurate staging and follow-up after treatment. Among the newer imaging technologies to be discussed here are digital mammography, computer-aided diagnosis (CAD), power Doppler ultrasound, magnetic resonance imaging (MRI), isotope imaging: methoxyisobutyl-isonitrile (MIBI) and positron emission tomography (PET). Semin Oncol 28:221-228. PMID- 11402432 TI - Minimal invasive staging for breast cancer: clinical experience with sentinel lymph node biopsy. AB - During the 1990s, considerable research and development resulted in reasonably reliable methods to target the set of lymph nodes most likely to contain metastases in patients with breast cancer. The methods of identification of these "sentinel nodes" (SNs) involve injection of a visual-based dye or a radioactive tracer. The tracer/dye enters the lymphatics and labels the SNs so that they can be selectively removed. SNs can be successfully identified in > or =90% of patients. In breast cancer patients with clinically negative lymph nodes, the accuracy of the SNs to predict whether any nodal metastases are present is > or =97%. The false-negative rate, however, ranges from 0% (in smaller series) to 11%. Clinical trials are in progress that will determine the long-term safety and predictive value of SN resection in patients with breast cancer. Successful application of SN surgery should allow elimination of conventional axillary lymphadenectomy in at least 75% of patients with breast cancer. Semin Oncol 28:229-235. PMID- 11402433 TI - Bone marrow staging for breast cancer: is it better than axillary node dissection? AB - Numerous studies have shown tumor cells in bone marrow to be a new and independent prognostic factor in primary breast cancer. The presence of such cytokeratin-positive or mucin-positive cells reflects the biology and systemic character of breast cancer much better than lymph node status. Axillary lymphadenectomy is associated with a considerable number of complications and is completely unnecessary in about 50% of cases (in node-negative patients). Whether axillary node dissection contributes to improved survival is highly controversial. However, since nearly all patients with primary breast cancer now receive adjuvant systemic therapy, the value of the classic prognostic factors must be discussed and re-evaluated. Much information can now be determined from primary tumor (HER-2/neu, etc). Tumor cell detection in bone marrow is a simple method that can be performed on an outpatient basis and that can be repeated if necessary (for monitoring therapy). The main disadvantage of the technique is that it has not been possible to standardize the laboratory methods and to find the ideal antibody--one that is not only able to recognize an epithelial cell, but which can also describe its metastatic potential. Semin Oncol 28:236-244. PMID- 11402434 TI - Postmastectomy locoregional radiotherapy: is it here to stay? AB - Postmastectomy radiotherapy (PMRT) clearly reduces the risk of locoregional recurrence for patients with invasive breast cancer with involved axillary lymph nodes. For many years it has been controversial whether or not it also decreases the risks of distant failure and, ultimately, death due to cancer, especially for patients receiving systemic therapy. There is now clear evidence that PMRT confers such benefits. However, published series vary substantially in describing the incidence of locoregional failure (LRF) following mastectomy with regards to particular patient subsets (such as those defined by the number of involved axillary nodes). Details of surgery and systemic therapy also affect this risk. This chapter will review the available data on these subjects, as well as their interpretation and clinical use. Much remains uncertain with regards to the benefits of PMRT in specific situations. Nonetheless, it appears that PMRT is indeed "here to stay." Semin Oncol 28:245-252. PMID- 11402435 TI - Breast cancer chemoprevention: current status and future directions. AB - Cancer prevention is a relatively young concept. Perhaps the greatest strides in cancer research that have been made in the prevention arena have been in breast cancer. In this article we examine the meaning of "prevention" and discuss several of the trials under way or completed, including the National Surgical Adjuvant Breast and Bowel Project (NSABP) Breast Cancer Prevention Trial. Semin Oncol 28:253-259. PMID- 11402436 TI - Tamoxifen to raloxifene and beyond. AB - Tamoxifen, a selective estrogen receptor modulator (SERM), is the treatment of choice for all stages of hormone-responsive breast cancer and has been shown to prevent breast cancer in high-risk women. Despite acting as an antiestrogen on the breast, tamoxifen has partial estrogenic effects on other target tissues. These partial estrogen agonistic actions produce beneficial effects on bones and the lipid profile in postmenopausal women. However, tamoxifen is associated with an increase in endometrial cancer. Additionally, its antiestrogenic effects in the central nervous system result in hot flashes in postmenopausal women. Raloxifene is another SERM approved for the prevention of osteoporosis in postmenopausal women. Like tamoxifen, raloxifene appears to prevent breast cancer in high-risk women and has not, to date, been noted to increase the incidence of endometrial cancer. The Study of Tamoxifen and Raloxifene will compare the effects of the two agents on breast cancer prevention and endometrial cancer risk. A number of new agents are being developed for breast cancer treatment and prevention and osteoporosis prevention. These include other SERMs, selective estrogen receptor downregulators (SERDs), and aromatase inhibitors. It is hoped that one of these new agents will be the ideal agent, acting as an antiestrogen on breast and endometrium while having estrogenic effects on bones, the lipid profile, and the central nervous system. Semin Oncol 28:260-273. PMID- 11402437 TI - Menopause and breast cancer: symptoms, late effects, and their management. AB - Menopause and breast cancer are common independent health events, and coincide in some women. This article will describe what is known about the impact of menopause on breast cancer survivors, focusing on short-term symptoms and other late effects. Common menopausal syndromes in breast cancer survivors will be examined, with suggestions for their management. Strategies for prevention of the late effects of menopause in breast cancer survivors will be considered. Semin Oncol 28:274-283. PMID- 11402438 TI - The evolving role of bisphosphonates. AB - Bisphosphonates, analogs of pyrophosphate, bind to bone at sites of active bone remodeling. In clinical settings of rapid bone turnover and/or excessive osteolytic activity, they have been shown to have beneficial clinical effects. These settings include Paget's disease of bone, osteoporosis from a variety of clinical causes, and malignant bone disease. Bisphosphonate inhibition of osteolysis in cancer has been shown to be effective therapy for malignancy associated hypercalcemia and as adjunctive therapy for the delay or prevention of cancer-related skeletal morbidity, including bone pain, pathologic fractures, and need for radiation therapy. Animal models of bone metastasis prevention by bisphosphonate treatment have provided the preclinical background for the adjuvant use of bisphosphonates in primary cancers. The success of these clinical trials has provided strong impetus for new research on bone disease and malignancy, as well as the development and testing of new and more potent bisphosphonates. Semin Oncol 28:284-290. PMID- 11402439 TI - Endocrine therapy in the treatment of metastatic breast cancer. AB - The goals of treating patients with metastatic breast cancer are to prolong survival, slow or halt disease progression, and enhance the patient's quality of life. In patients with estrogen receptor (ER)-positive cancers that are not progressing rapidly, endocrine therapy is generally the first treatment option. If a patient initially responds to an endocrine agent and then progresses, another endocrine agent may still provide benefit. Tamoxifen has been used as first-line therapy for metastatic breast cancer for many years. Until recently, no other endocrine agent has shown superiority to tamoxifen in this setting. The nonsteroidal aromatase inhibitors, anastrozole and letrozole, have been widely accepted as second-line therapy after failure of tamoxifen; they have replaced megestrol acetate in this setting. Recently, anastrozole was shown to have at least equivalent efficacy and a superior side effect profile compared with tamoxifen for treating postmenopausal women in the first-line setting. Thus, this aromatase inhibitor has become a viable option for first-line therapy in postmenopausal women. Trials of letrozole in this setting are nearing completion. Exemestane has been shown to be an effective second-line agent and to have at least some efficacy as a third-line agent even after failure of a nonsteroidal aromatase inhibitor. Results are anxiously awaited from trials of new endocrine agents including the first member of a new class of endocrine agent, the estrogen receptor downregulator class. Semin Oncol 28:291-304. PMID- 11402440 TI - Incorporating genomics into the cancer clinical trial process. AB - The effectiveness of current chemotherapeutic approaches for the treatment of solid tumors has reached a near plateau, suggesting we are nearing the limit of cytoreduction. It is hypothesized that this may be due to "subset effect," and that drugs administered according to responses predicted for particular subgroups within the population being treated could overcome what appears to be a limit of cytoreduction. However, the clinical trial process, as currently structured, prevents efficient discovery and validation of predictive markers of treatment response. An alternative process is proposed, based on preoperative therapy and high-throughput multiplexing of markers to provide a built-in, unbiased discovery and validation process for predictive markers. Semin Oncol 28:305-309. PMID- 11402441 TI - [A study of Wilson's disease gene encoded products and gene mutations]. AB - OBJECTIVE: To investigate the pathogenesis of Wilson's disease(WD). METHODS: Hepatocytes were isolated from WD patients' bioptic hepatic samples and cultured in vitro; WD proteins, the gene putative encoded products, were detected by SDS PAGE in conjunction with Western blotting in liver samples of three patients and two controls. Their genomic DNAs were analyzed by means of direct DNA sequencing of WD gene (ATP7B) on exon 8. RESULTS: The WD proteins lanes from two WD patients were found to be much weaker than that from the control, from which one WD patient was proven as heterozygote of 778 position CGG-->CTG(Arg778Leu) and 770 position CTC-->CTG change of ATP7B. CONCLUSION: WD is highly heterogeneous in clinical manifestations and inheritance pattern. Abnormally expressed putative WD proteins in WD patients might be the results of ATP7B mutations, and the study of ATP7B products would help to probe into the pathogenesis of WD. PMID- 11402442 TI - [Study of susceptibility loci located within Xp11 in attention deficit hyperactivity disorder]. AB - OBJECTIVE: To detect the genetic relationship between monoamine oxidase(MAO) A type gene and attention deficit hyperactivity disorder(ADHD) in Chinese. METHODS: The haplotype-based haplotype relative risk(HHRR) and the transmission disequilibrium test(TDT) methods were used to analyze the genetic association and linkage in 60 ADHD children and their parents. RESULTS: In this sample were found significant association (chi(2)=4.90, P<0.05) and linkage (chi(2)=4.84, P<0.05) between the MAOCA 114bp allele and DSM-III-R-diagnosed ADHD in trios composed of father, mother and affected offspring. CONCLUSION: The above results suggested that ADHD was associated and in linkage with MAO A gene, and the susceptibility loci might reside in chromosome Xp11 for ADHD. PMID- 11402443 TI - [Analysis on genetic polymorphism of 13 STR loci on chromosomes 11 and 19 in Chinese Hans]. AB - OBJECTIVE: To analyze the genetic polymorphism of 8 STR loci on chromosome 11 and 5 STR loci on chromosome 19 in Chinese Hans. METHODS: Polymerase chain reaction single strand length polymorphism(PCR-SSLP) was used to genotype 100 randomly selected individuals of the Han nationality at 8 STR loci(D11S1984, D11S1999, D11S1392, D11S1985, D11S2002, D11S1986, D11S4464 and D11S2359) on chromosome 11 and 5 STR loci(D19S247, D19S714, D19S433, D19S246, D19S254) on chromosome 19. RESULTS: Eight alleles and 26 genotypes, 9 alleles and 15 genotypes, 6 alleles and 16 genotypes, 12 alleles and 40 genotypes, 6 alleles and 19 genotypes, 12 alleles and 48 genotypes, 8 alleles and 20 genotypes, 7 alleles and 13 genotypes were observed at D11S1984, D11S1999, D11S1392, D11S1985, D11S2002, D11S1986, D11S4464 and D11S2359. The heterozygosities for the 8 STR loci were 87%, 68%, 73%, 92%, 71%, 86%, 75% and 71%, respectively. Ten alleles and 19 genotypes, 10 alleles and 26 genotypes, 10 alleles and 24 genotypes, 11 alleles and 29 genotypes, 8 alleles and 18 genotypes were observed at D19S247, D19S714, D19S433, D19S246, D19S254. The heterozygosities for the 5 STR loci were 63%, 82% 72%, 81% 74%, respectively. CONCLUSION: The distribution of allele frequencies of 8 STR loci on chromosome 11 and 5 STR loci on chromosome 19 was consistent with the Hardy-Weinberg equilibrium and the highly genetic polymorphism was observed in Chinese Han population. PMID- 11402444 TI - [Reduction of FHIT gene expression in primary lung cancer: relationship with the proliferation and apoptosis of tumor cell]. AB - OBJECTIVE: To investigate the role of FHIT(fragile histidine triad) gene in oncogenesis and progression of human lung cancer and explore the relationship of FHIT gene expression with the proliferation and apoptosis of tumor cells. METHODS: The expression of FHIT gene and that of Ki-67 were detected in 166 lung cancer samples and 37 lung benign lesion tissues as control by immunohistochemistry, and the apoptotic level of tumor cell was detected by terminal deoxynucl neotidyl transferase mediated dUTP nick end labelling(TUNEL). RESULTS: (1)The expression level of FHIT gene was found to be significantly lower in lung cancer tissues than in benign lesion tissues (P<0.01); (2)The expression level of FHIT gene was closely related to histological classification, cancer cell differentiation, P-TNM stages and lymph node involvement in lung cancer patients (P<0.05); (3)Lasting and heavy smoking might be one of the important reasons of reduction of FHIT gene expression in lung cancer patients; (4)The postoperative survival time of patients in high FHIT expression group was significantly longer than those in low expression group (P<0.05); (5)The reduction of FHIT gene expression might be related with the over-proliferation and suppression of apoptosis of lung cancer cell(P<0.01). CONCLUSION: FHIT gene may play a role in the regulation of proliferation and apoptosis of lung cancer cell. And reduction of FHIT gene expression may be related to the pathogenesis and development of lung cancer. PMID- 11402445 TI - [Study of a familial insertional translocation involving chromosomes 1 and 7 by using fluorescence in situ hybridization]. AB - OBJECTIVE: To determine the karyotype of a case with a history of spontaneous abortion and terminal deletion by using of conventional G-banding method and search the cause of insertional translocation of chromosomal terminal region. METHODS: Fluorescence in situ hybridization (FISH) technique was performed to analyze the case by using whole chromosome 7 painting probe and subterminal probe of 7q36-->qter which was generated by chromosome microdissection technique. RESULTS: The case was a carrier with a very rare insertional translocation involving chromosomes 1 and 7. The region of chromosome 7q36-->qter was not inserted into chromosome 1. The abnormal chromosome was inherited from her mother. CONCLUSION: The present authors provided an experiment evidence that in this case the chromosome insertional translocation including the terminal region was still a three breakage rearrangement and the terminal deletion found by cytogenetics should be an interstitial deletion. Combining with chromosome microdissection, FISH technique is a powerful diagnostic method for detecting the chromosome structural abnormality. PMID- 11402446 TI - [Telomere length and DCC gene mRNA expression of human large intestine cancers]. AB - OBJECTIVE: To evaluate the role of telomere and DCC in tumor transformation and progression. METHODS: Telomere length and DCC gene mRNA expression were examined by southern blot hybridization and RT-PCR analysis in 46 adenomas of large intestine, 62 cancers of large intestine and corresponding normal mucosa. RESULTS: Shortening of the telomere was present in the tissues of 41.3% of the adenomas and 53.2% of the cancers, and their average TRF lengths were significantly shorter than those of corresponding normal mucosa(P<0.05, P<0.01), whereas the telomere elongation was only detected in 4.4% and 6.5% of the adenomas and cancers respectively. In addition, the average telomere length in colon carcinomas was also shorter than that in rectal carcinomas. Moreover, the average telomere lengths of the colorectal cancer mucosa became shorter with age. The rates of DCC mRNA expression deletion were 34.8% and 62.9% in the tissues of adenomas and cancers respectively. The DCC mRNA expression deletion occurred more frequently in poorly differentiated and Dukes C, D carcinomas than in well differentiated and Dukes A, B carcinomas (P<0.05, P<0.01). However, no significant correlation was found between the length of telomere and the deletion of DCC mRNA expression in the cancers of large intestine. CONCLUSION: The telomere shortening and DCC mRNA deletion may represent the biologic behavior of transformation and development of the large intestine cancers. PMID- 11402447 TI - [Intron 44 is not the most unstable intron in the "central deletion hot spot" of dystrophin gene]. AB - OBJECTIVE: To understand the distributional characteristics of dystrophin gene deletion breakpoints in "central deletion hot spot" and analyze the instability of introns 44-51 after excluding the effect of intron's length. METHODS: Fifty nine Duchenne/Becker muscular dystrophy(DMD/BMD) patients were detected by polymerase chain reactions with the primers to amplify exons 44-52 of dystrophin gene. The amount of actual breakpoints, expected breakpoints according to its length, and the ratios of actual breakpoints to expected values(A/E) for introns 44-51 were calculated respectively. RESULTS: In "central deletion hot spot", about 30.8% of breakpoints fell in intron 44, about 23.1%, 17.9%, 10.3%, 10.3% of breakpoints fell in introns 50,51, 45, 48, respectively. But the amount of actual breakpoints of intron 44 was less than that of expected breakpoints according to its length, the ratio of A/E was 0.7. The amount of actual breakpoints of introns 48, 50, 51, 45 were more than that of length expected value. The ratios of A/E were 2.7, 2.0, 1.9, 1.1, respectively. CONCLUSION: Intron 44 is more stable than the whole molecular region of "central deletion hot spot". Introns 48, 50 and 51 are comparatively instable in "central deletion hot spot". PMID- 11402448 TI - [Association analysis of attempted suicide and 5-HT2A receptor gene]. AB - OBJECTIVE: To examine the relationship of attempted suicide with A-1438G and T102C polymorphisms of 5-HT2A receptor gene. METHODS: Genotypes and allele frequencies of A-1438G and T102C polymorphisms of 5-HT2A receptor gene in 149 attempted suicides and 190 normal controls were examined by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). RESULTS: No differences in genotypes and allele frequencies of A-1438G and T102C polymorphisms of 5-HT2A receptor gene were observed between attempted suicides and the controls. However, an association between male attempted suicides and genotype(GG) of A-1438G polymorphism was found. CONCLUSION: These results suggested that liability to suicide behavior might be associated with A-1438G polymorpyhism of 5-HT2A receptor gene in male subjects. PMID- 11402449 TI - [Comparison of denaturing high performance liquid chromatography with direct sequencing in the detection of single nucleotide polymorphism]. AB - OBJECTIVE: Investigate the sensitivity and accuracy of the denaturing high performance liquid chromatography(DHPLC) technique for the detection of single nucleotide polymorphism(SNP). METHODS: Forty-one samples were detected by both DHPLC and direct sequencing. RESULTS: The comparison demonstrated that DHPLC detected all heterozygous sequences found by direct sequencing. No false-positive signals were seen in the cases of homozygous sequences. Furthermore, no false negative results were ever obtained with heterozygous mutations or polymorphisms, or both. CONCLUSION: DHPLC is a potent method for SNP identification especially SNP typing in large scale screening. PMID- 11402450 TI - [Genotype polymorphism and its implications of mannose-binding protein allele in 5 Chinese nationalities]. AB - OBJECTIVE: To detect the genotypes and sequences of the exon 1 of human mannose binding protein (MBP) allele in 5 Chinese nationalities. METHODS: The genotypes of MBP gene of 5 Chinese nationalities were detected by polymerase chain reaction restriction fragment length polymorphism(PCR-RFLP). The exon 1 of the MBP gene of 22 Chinese Hans was analyzed by using ABI 310 genetic analyzer. RESULTS: The DNA sequences of exon 1 of Chinese MBP gene were acquired. The allele frequencies of the codon 54 of the MBP gene (MBP-54) of 5 Chinese nationalities were 0.181(Hans), 0.128(Uygurs), 0.181(Mongols), 0.179(Tibetans) and 0.181(Yis). The allele distribution for MBP-54 mutation of 5 Chinese nationalities was in good agreement with Hardy-Weinberg equilibrium. Compared with the Hans, Uygurs had a lower MBP-54 mutation rate. There were no differences in the allele frequencies between the chronic hepatitis B patients and health controls in Chinese Hans. The mutations of the codons 52 and 57 were not detected in this study. CONCLUSION: A higher prevalence of MBP-54 mutation was found in 5 Chinese nationalities, MBP-54 mutation was not associated with the persistence of hepatitis B. PMID- 11402451 TI - [Study on the relationship between methylenetetrahydrofolate reductase gene C677T mutation and coronary heart disease]. AB - OBJECTIVE: To investigate whether methylenetetrahydrofolate reductase (MTHFR) gene C677T polymorphism is linked with coronary heart disease (CHD). METHODS: Transmission/disequilibrium test(TDT), sib transmission/disequilibrium test(STDT), and sibship disequilibrium test(SDT) were used. Forty-five CHD pedigrees with at least one CHD patient in the first degree relatives of probands were recruited from Oct. 1998 to Feb. 1999. Among those recruited were 21, 2 and 22 pedigrees with the genotypes of both parents known, one parental genotype unknown and both unknown, respectively. MTHFR genotype was measured by PCR-RFLP technique. RESULTS: Neither the TDT for 23 nuclear families with at least one parental genotype known or the STDT and SDT for 40 sibships found significant difference between the transmitted and untransmitted MTHFR gene 677T allele distributions. CONCLUSION: The above results suggest that MTHFR gene 677T allele is probably not linked with CHD in Chinese population. PMID- 11402452 TI - [Study on the association of transporter associated with antigen processing gene with ankylosing spondylitis and HLA-B27 in Han nationality]. AB - OBJECTIVE: To investigate the association between transporter associated with antigen processing(TAP) gene and ankylosing spondylitis(AS) as well as HLA-B27. METHODS: PCR-SSO was used to analyze the frequencies of TAP alleles and variant amino acids of TAP in 48 patients (B27(+)) with AS and 123 normal individuals (B27(+) or B27(-)). RESULTS: The phenotypes of Ile/Ile, Asp/Asp were over presented for TAP1 and Val/Val, Ala/Thr, Stop/Stop for TAP2 in Han nationality. For each TAP there were four alleles: TAP1*0101, TAP1*0201, TAP1*0301, TAP1*0401, and TAP2*0101, TAP2*0102, TAP2*0201, TAP2*0202. In the research subjects, hybridization blanks accounted for 9.9%(17/171) of TAP1 and 15.8%(27/171) of TAP2. No difference was found in the distribution of TAP alleles between the AS patients(B27(+)) and normal controls(B27(+) or B27(-)). In the patients group the frequency of phenotype in codon of 333 (Val/Val) was significantly decreased as compared with that of the healthy controls(B27(+))(P<0.05). In addition, the frequency of phenotype Ala/Ala(665) was significantly decreased and that of Ala/Thr was significantly increased in normal controls(B27(-)), compared with those in normal controls(B27(+))(P<0.05). CONCLUSION: It is possible that there are new TAP alleles in Han nationality. No association was found between TAP alleles and AS. And the homozygosity for codon 333 which is Val/Val seems to offer resistance to AS among those HLA-B27(+) individuals. In normal individuals, negative correlation was found between heterozygote Ala/Thr and B27, and possibly there was positive correlation between phenotype Ala/Ala and B27. PMID- 11402453 TI - [Gene diagnosis of facioscapulohumeral muscular dystrophy]. AB - OBJECTIVE: Perform gene diagnosis for Chinese facioscapulohumeral muscular dystrophy(FSHD). METHODS: Digest genome DNA with restriction enzymes EcoR I only and EcoR I associated with Bln I. Use 0.6% agarose gel electrophoresis and Southern blotting hybridization with probe P13E11. RESULTS: For FSHD patients, the sizes of EcoR I + Bln I/P13E11 DNA fragments ranged from 15kb to 33kb. For normal controls, they were over 41kb. Two presymptomatic patients were found. CONCLUSION: It is feasible to perform gene diagnosis and presymptomatic diagnosis for most Chinese FSHD patients by Southern blotting hybridization with probe P13E11, following double digestion of genome DNA with restriction enzymes EcoR I and Bln I. PMID- 11402454 TI - [Rapid detection of the common alpha-thalassemia-2 determinants by PCR assay]. AB - OBJECTIVE: Most frequent molecular lesions of alpha-thalassemia-2 determinants are deletions of one alpha-globin gene. Satisfactory PCR methodologies for detecting the deletions are required for molecular diagnosis and genetic screening since there was no internal control in most published PCR-based strategies. The purpose of this study is to develop a reliable PCR protocol specific for the two common alpha-thalassemia-2 determinants with internal control. METHODS: The multiple repeat elements and the high GC-content of the alpha-globin locus impose severe limitations on designing suitable primers and optimizing stable conditions for PCR. In this study, two multiplex PCR systems were successfully set up. One was designed to detect the rightward deletion ( alpha(3.7)/) with two pairs of primers including one newly optimized pair for amplification of the internal standard to indicate the success of failure of PCR amplification. The other, to the leftward deletion(-alpha(4.2)/) with three primers, which were designed according to the newly sequenced data of the alpha(4.2) and HbQ-alpha(4.2) deletions in this lab(Genbank Accession No. AF221717). In the PCR system, one is used as a common upstream primer and the other two are used as specific downstream primers for typing the normal allele and the deletion one, respectively. RESULTS: Easily interpretable, unambiguous amplifications were observed by using the multiplex PCR systems for the detection of the two common alpha-thalassemia-2 determinants. The three or four primers were run in the same tube under the same condition and both of these two systems could be used at the same thermal cycle parameters. For typing the rightward deletion, a mutant-specific amplification of 1.7 kb and a 1140 bp amplified band as a normal and system control were produced. For typing the leftward deletion, two PCR-amplified bands, a 956 bp fragment specific for a -alpha(4.2) gene and a 1140 bp one for a normal allele were found. CONCLUSION: Two sets of PCR systems with internal controls for detecting the most common two alpha-thalassemia-2 determinants have been established and may be suitable for molecular diagnosis and population screening. PMID- 11402455 TI - [Cyclin-dependent kinase 4 inhibitor a and tumor]. AB - INK4a/ARF locus encodes at least two products, P16(INK4a) and P19(ARF), which are very important in controlling cell-cycle arrest and apoptosis. Many human cancers have been found to be highly related to the mutations at this locus. Here is a review of its structure, function and role in tumor suppression. PMID- 11402456 TI - Intra-pocket antibiotic therapy using resorbable and non-resorbable slow-release devices containing tetracycline. AB - Since it is a disease mainly caused by plaque--an aggregate of various bacteria- periodontal disease can be considered a local infection. Thus, it has seemed reasonable to utilize antibiotics to suppress the intrapocket bacteria, specifically or nonspecifically. When antibiotics are administered orally, however, massive doses over a prolonged period of time are needed to attain a therapeutic effect. This increases the risk of adverse reactions as well as developing resistant strains of bacteria. To overcome these problems, local drug delivery systems (LDDS) were devised to combat the local infection. However, the intrapocket antibiotic delivery systems have yet to be fully evaluated for clinical effectiveness; to prove the therapeutic effectiveness of locally administered antibiotics, the drug must reach the base of the periodontal pocket and the effective concentration of the antibiotic against the pathogenic bacteria must be maintained for a long time. This concise review presents with figures, tables, and a comprehensive list of references the many studies which have used the various tetracyclines as LDDS to treat periodontal disease. PMID- 11402457 TI - The effect of a collagen membrane in regenerative therapy of two-wall intrabony defects in dogs. AB - Guided tissue regeneration that supports the periodontal ligament and bone cells in achieving healthy attachment between teeth and alveolar bone following periodontal therapy has been repeatedly described in the literature. The aim of the present study was to assess the effect of an absorbable collagen membrane used in guided tissue regeneration procedures in two-wall intrabony defects. For this purpose, periodontal defects were surgically created around mandibular teeth in nine dogs. In a randomly chosen quadrant in each dog, a collagen membrane was shaped to cover the interproximal bone defect and adjacent root surface. No collagen membrane was placed over the control defects. Block biopsies of test and control sites were obtained from three dogs at 30 days, three dogs at 60 days, and three dogs at 90 days after the procedures. Histomorphologic and histometric evaluations were performed. We observed that both collagen membrane treated and control defects demonstrated similar amounts of new attachment and bone. However, gingival recession and postoperative keratinized tissue loss were observed in most of the sites. Although there was a tendency towards new attachment in both groups, the gingival tissue loss due to recession led to limited regeneration. PMID- 11402458 TI - Osseous repair with and without the use of bone grafts: long-term follow-up. PMID- 11402459 TI - [Functional anatomy and evaluation of hand function]. PMID- 11402460 TI - [Drug administration routes for the hand and wrist]. PMID- 11402461 TI - [Imaging studies of the hand and wrist]. PMID- 11402462 TI - [Role of arthroscopy of the wrist]. PMID- 11402463 TI - [Dystonia of the hand. Semiological analysis and management]. PMID- 11402464 TI - [Wrist injuries]. PMID- 11402465 TI - [Avascular necrosis of the carpal bones]. PMID- 11402466 TI - [Osteoarthritis of the fingers and trapeziometacarpal joints]. PMID- 11402467 TI - [Osteoarthritis of the fingers: challenges for the surgeon]. PMID- 11402468 TI - Survival and patterns of relapse following treatment of small-cell lung cancer at a community cancer center. AB - BACKGROUND: Although data from previously published trials have suggested an incremental improvement in response rate and survival in patients with small-cell lung cancer, few of these trials originate from private practice/community cancer center settings. The outcomes of patients with small-cell lung cancer treated in such a practice setting are reviewed and compared to prior studies with particular attention to the potential role of prophylactic cranial irradiation (PCI). METHODS: One hundred eighty patients treated between 1988 and 1998 comprise the basis of this analysis. In each individual, the specifics of therapy were recorded as well as the best response, sites of relapse, and survival. RESULTS: The disease was limited in extent (LD) in 99 patients (55%) and extensive (ED) in 81 (45%). The overall/complete response rates for LD patients and ED patients respectively were 90.9%/72.7% and 61.7%/18.5%. Median survivals for LD and ED patients were 20.2 and 9.59 months respectively. The one-year, two year, and five-year actual survival rates for the entire group were 59.1%, 22.2%, and 8.7%. Central nervous system relapses occurred in 42.9% of this largely nonprophylactic cranial irradiation treated patient population. The central nervous system was the only site of relapse in 13.6%. CONCLUSIONS: The outcomes of patients treated in this private practice/community cancer center setting are similar to those previously published from single university and multi institutional studies. Although these data support the use of cranial irradiation in patients who achieve a complete response, the impact of such an intervention will remain relatively insignificant until there is an improvement in systemic therapy. PMID- 11402469 TI - Definitive surgical trauma care live porcine session: a technique for training in trauma surgery. AB - BACKGROUND: A new Definitive Surgical Trauma Care course was developed to educate surgeons in operative management of injuries. The course consists of an interactive CD-ROM and a live porcine animal laboratory. METHOD: A five-hour session was conducted. Penetrating injuries to stomach, bowel, diaphragm, spleen, pancreas, kidney, ureter, inferior vena cava, liver, and heart were created by the senior surgeon and managed by the junior surgeon. Participants rated their expertise in 26 maneuvers pre- and post-lab. The evaluation scale used was: no prior experience; able to perform skill with assistance; proficient at procedure; able to teach procedure to another surgeon. RESULTS: In 26 procedures, a maximum score of 78 was possible. There was an increase from pre- to post-session scores of 22.6 for PGY-4 residents (n = 3); 23.3 for PGY-5 residents (n = 4); 11.25 for fellows (n = 4); and 0 for attendings (n = 4). CONCLUSION: The operative animal session had the greatest educational benefit among surgeons without formal training in trauma surgery. By exposing them to a range of trauma-induced surgical conditions, the DSTC course develops their operative repertoire and should increase their effectiveness in managing trauma patients. PMID- 11402470 TI - Dexmedetomidine, a novel alpha 2-adrenergic agonist for short-term sedation in the intensive care unit. PMID- 11402471 TI - The autopsy and medical fallibility: a historical perspective. AB - The three major factors causing the decline of nationwide autopsy rates to 0% to 5% are presented in historical perspective. One is the evolution of a perception by the public and doctors that infallible medical practice is a realistic goal in spite of the inevitability of unavoidable errors. Another is the structure of our tort-claim/liability insurance system that results in holding physicians culpable and financially responsible for injuries to patients by unavoidable errors. The third factor is the reluctant neglect of the autopsy by pathologists whose higher priorities are patient-care services. Inadequate funding for services and facilities to offset the lack of any income production has heightened neglect of the service. Remedies are presented for consideration. PMID- 11402472 TI - "Sum ergo cogito". PMID- 11402473 TI - "Can't we all just get along? PMID- 11402474 TI - Factors affecting the sterility of work areas in barrier isolators and a biological safety cabinet. AB - Factors affecting the sterility of the work area in barrier isolators and a biological safety cabinet (BSC) were studied. A study in a mother-and-child tertiary care teaching hospital in Canada examined the effects of a range of variables on the sterility of work areas in four barrier isolators and a standard BSC. The variables consisted of type of equipment (barrier isolator or BSC), day of the week (Monday through Thursday), time of day (0800-1000, 1000-1200, 1200 1400, and 1400-1600), sampling site (16 surfaces and 5 air sedimentation zones), type of product prepared (antimicrobial, total parenteral nutrient solution, etc.), cleaning procedure (before or after primary cleaning), and level of product preparation activity (none to intense). A total of 657 surface and air sedimentation samples, 327 plated onto Trypticase soy agar (TSA) and 330 onto Sabouraud dextrose agar (SAB-D), were taken during a 20-day period. Thirty-three (5%) of the samples yielded microbial growth when cultured (24 on TSA and 9 on SAB-D). A total of 74 isolates were identified, including Bacillus, Staphylococcus, Penicillium, Micrococcus, Corynebacterium, and Mucor species. Single-variable analysis showed that sampling site, sample type, the time of day samples were taken, and the types of equipment contributed significantly to microbial growth in the samples taken. Several variables were associated with microbial growth in samples from the work areas of barrier isolators and a BSC. More study is needed to compare BSCs and barrier isolators with respect to sterility. PMID- 11402475 TI - Alternative therapy use and adherence to antihyperlipidemic drugs in a lipid clinic. PMID- 11402476 TI - Stability of amphotericin B in an extemporaneously compounded oral suspension. PMID- 11402477 TI - Japanese pharmaceutical education: implications for Americans teaching Japanese pharmacy students. PMID- 11402478 TI - Changes in home care accreditation: enhancing patient care at alternative sites. PMID- 11402479 TI - Marketing a hospital-based drug information center. PMID- 11402480 TI - Individualized health care and the pharmaceutical industry. PMID- 11402481 TI - Yellowing of metabisulfite-containing propofol emulsion. PMID- 11402482 TI - Yellowing of metabisulfite-contaning propofol emulsion. PMID- 11402483 TI - Scope of home care guidelines. PMID- 11402484 TI - Adverse drug events in children analyzed. PMID- 11402485 TI - Oral prodrug of ganciclovir approved for CMV retinitis. PMID- 11402486 TI - Changes recommended in adverse event reporting on dietary supplements. PMID- 11402487 TI - Severe depression does not respond to St. John's wort, study says. PMID- 11402488 TI - Prescription drug misuse highlighted as national problem. PMID- 11402489 TI - Privacy regulations go into effect, with modifications to come later. PMID- 11402490 TI - Developing and maintaining up-to-date training for pharmacy technicians. PMID- 11402491 TI - Achieving provider recognition. PMID- 11402492 TI - Kelsey's story. PMID- 11402493 TI - Sharing responsibility for patient safety. PMID- 11402494 TI - Pantoprazole. AB - The pharmacology, pharmacokinetics, clinical efficacy, adverse effects, and dosage and administration of pantoprazole are reviewed. Pantoprazole is a gastric hydrogen-potassium adenosine triphosphatase (H+/K(+)-ATPase) inhibitor. It shares the same core structure as other currently available proton-pump inhibitors (PPIs). The FDA-labeled indication is the short-term treatment of erosive esophagitis. PPIs act by selectively inhibiting H+/K(+)-ATPase in the secretory canaliculus of the stimulated parietal cell. Understanding the pharmacodynamics of PPIs is more relevant than knowing their pharmacokinetics, since the duration of action depends on the rate of de novo proton-pump regeneration, not the duration of drug circulation in the body. Pantoprazole is well absorbed, undergoes little first-pass metabolism, and has an absolute bioavailability of approximately 77%. Pantoprazole has been evaluated in more than 100 clinical trials involving more than 11,000 patients. It is effective in treating erosive esophagitis and duodenal and gastric ulcers. It is also effective as adjunctive treatment with antimicrobials in patients infected with Helicobacter pylori. Pantoprazole has been shown to control acid production in Zollinger-Ellison syndrome. Pantoprazole is well tolerated. The most commonly reported adverse effects are headache, diarrhea, and abdominal pain. The recommended oral dosage for erosive esophagitis is 40 mg once a day for up to eight weeks. The recommended i.v. dose is 40 mg given over 15 minutes once a day in patients with gastroesophageal reflux disease who are unable to take oral medication. Pantoprazole appears to be as safe and effective as other PPIs in acid-related disorders. PMID- 11402495 TI - Effect of peptide-carrier coupling on peptide-specific immune responses. AB - Synthetic peptides are covalently linked to immunogenic carrier proteins to enhance the anti-peptide immune response. To investigate whether the method of conjugation influences the immune response, we evaluated two distinctly different choices of linker for a peptide-carrier construct. HPG-30, a synthetic peptide derived from the p17 gag protein of human immunodeficiency virus 1, was covalently linked to keyhole limpet hemocyanin by either glutaraldehyde or a maleimide ester. Glutaraldehyde linkage enhanced the anti-peptide antibody and native protein response compared to maleimide. The maleimide-linked conjugate was more effective at inducing a peptide-specific cellular response. Thus, manipulation of the conjugation method can modify the magnitude and character of the immune response to a synthetic peptide vaccine. PMID- 11402496 TI - Signal transduction via both human low-affinity IgG Fc receptors, Fc gamma RIIa and Fc gamma RIIIb, depends on the activity of different families of intracellular kinases. AB - In contrast to IgG Fc receptor II (Fc gamma RIIa), the function of Src-family kinases, Syk and phosphoinositide 3-kinase (PI3K) in signal transduction of glycosylphosphatidylinositol-anchored Fc gamma RIIIb has not been analyzed in detail. Therefore pharmacological inhibitors were used to define the role of specific kinases in signalling of Fc gamma RIIa and Fc gamma RIIIb in myeloid cells. We demonstrate that the broad tyrosine kinase inhibitor genistein, the Src family kinase inhibitor PP2, and the Syk kinase inhibitor piceatannol inhibit [Ca2+]i rise induced by both low-affinity Fc gamma R in neutrophils. Genistein and PP2 additionally prevent Fc gamma R-triggered hydrogen peroxide generation. In contrast, wortmannin, a PI3K inhibitor, which blocks Fc gamma RIIIb activation completely, abolishes Fc gamma RIIa-mediated [Ca2+]i flux only in the beginning. In addition, herbimycin A, a further specific inhibitor of Src-family kinases leads to a delayed Fc gamma RIIa-induced [Ca2+]i rise in THP-1 cells. In summary, our data demonstrate differences between both low-affinity IgG Fc receptors, and provide evidence for an essential role of Src-family kinases, Syk and PI3K in Fc gamma RIIIb-mediated signalling. PMID- 11402497 TI - Induction of apoptosis by 7,8-dihydroneopterin: involvement of radical formation. AB - Interferon-gamma is a cytokine released in large amounts during cell-mediated immune response. It induces the expression of proinflammatory cytokines and enhances macrophage capacity to secrete reactive oxygen intermediates and the pteridines neopterin and 7,8-dihydroneopterin. To assay the role of these pteridines in the immune system several studies were performed. Thereby, 7,8 dihydroneopterin was found to induce apoptosis in T lymphocytes. In this study we report that caspases are involved in 7,8-dihydroneopterin-mediated apoptosis in Jurkat T cells. In connection with this result we found that 7,8-dihydroneopterin can increase Fas ligand expression detected in Western blot analysis and promoter reporter assays. Antioxidants potently reduced the effect of 7,8-dihydroneopterin on Fas ligand promoter activation suggesting an involvement of oxidative stress. In further investigations, ESR-measurements were performed to evaluate the role of 7,8-dihydroneopterin in the formation of radicals. We found that the pteridine in combination with the spin trap DMPO induces the production of DMPO-OH spin adducts. This reaction was sensitive to the presence of chelated metal ions and could completely be blocked by the addition of superoxide dismutase. These data suggest that 7,8-dihydroneopterin in aqueous solution leads to the formation of .OH radicals via generation of superoxide anion. We hypothesize that an overproduction of radicals caused by high levels of 7,8-dihydroneopterin is likely to be responsible for the pro-apoptotic effects observed in cell cultures and possibly contributes to the pathogenesis of diseases involving immune activation and elevated concentrations of neopterin-derivatives. PMID- 11402498 TI - Neopterin production in SCID mice injected with human peripheral blood mononuclear cells. AB - Intraperitoneal transfer of peripheral blood mononuclear cells (PBMC) from human EBV+ donors into severe combined immunodeficiency (SCID) mice is a suitable model for studying some aspects of lymphomagenesis and immune activation. Neopterin is a soluble immune marker which was found to be a useful indicator for immune activation processes in humans, e.g. to monitor immunological complications in allograft recipients or to predict prognosis in HIV-infected individuals. In contrast, this pteridine compound is normally synthesized in murine organism in only very low amounts. The measurement of neopterin concentrations in serum and urine should be feasible in SCID mice reconstituted with human PBMC. In this study, we examined the usability of this experimental model for monitoring human T cell activation by neopterin measurements. The production of neopterin by SCID mice after injection of freshly isolated human PBMC, purified B or T cells and cultured Epstein-Barr virus (EBV)+ lymphoblastoid cells (LCL) was determined. It was found that neopterin can be detected early after injecting SCID mice with PBMC, whereas injection of purified human T or B cells did not result in neopterin production. Highest neopterin levels were detected in mice treated with LCL cells when developing lymphoma. We discuss the possible sources of neopterin along this process and its usefulness in this model. PMID- 11402499 TI - Unexpected anti-alpha GalNAc antibodies in alpha-galactosyl transferase-deficient mice: complex relationship between genotype and the natural antibody repertoire. AB - Mice lacking the alpha-galactosyl transferase gene (GalT(-/-) mice) have been used extensively as a model for xenotransplantation. Unlike wild type (WT) mice, GalT(-/-) mice do not produce Gal alpha 1-3Gal and are known to produce natural IgM specific for Gal alpha 1-3Gal, as do humans and higher primates. In addition to natural anti-Gal alpha 1-3Gal IgM in GalT(-/-) mice, we identified natural IgM which bound alpha-N-acetylgalactosamine (alpha GalNAc) but not Gal alpha 1-3Gal or blood group A. Although unexpected, these antibodies were expressed at 10-fold greater concentrations in GalT(-/-) mice than in WT mice. One explanation for this unexpected observation is that the production of natural antibodies is affected by self-antigen(s) that are similar but not identical to targets recognized by the natural antibody. Thus, the natural humoral immune system may be unresponsive to "near-self" antigens even though the individual is not tolerant to those antigens. Another explanation for the unexpected results is that there may be unanticipated and uncharacterized differences between GalT(-/-) mice and WT mice. These studies underscore the need to extensively characterize phenotypes in KO mice and indicate that the relationship between genotype and the natural immune repertoire can be complex. PMID- 11402500 TI - Engagement of CD147 molecule-induced cell aggregation through the activation of protein kinases and reorganization of the cytoskeleton. AB - CD147 is a broadly expressed cell surface molecule of the immunoglobulin superfamily whose expression is up-regulated upon T cell activation. Engagement of CD147 by CD147 monoclonal antibodies (mAbs) has been shown to induce homotypic aggregation of U937 cells. To study intracellular signal transduction induced by the engagement of CD147 molecules, protein kinase C (PKC) and protein tyrosine kinase (PTK) inhibitors were used to inhibit cell aggregation. The results indicated that a PKC inhibitor, sphingosine, and a PTK inhibitor, herbimycin A, inhibited CD147 mAb-induced cell aggregation in a dose-dependent manner. In contrast to herbimycin A, a PTK inhibitor, genistein, enhanced cell aggregation. This discrepancy may be due to the differential effect of herbimycin A and genistein on the target cells. Effect of actin filament polymerization blocking agent, cytochalasin B, was also studied and it was found that cytochalasin B completely inhibited CD147 mAb-induced cell aggregation. This result implied that U937 cell aggregation induced by CD147 mAbs is associated with cytoskeleton reorganization. Thus, our observations suggest that cell aggregation induced by the engagement of CD147 with specific mAbs depend upon the activation of protein kinases and a functional cytoskeleton. PMID- 11402501 TI - Enhancement of complement-mediated lysis by a peptide derived from SCR 13 of complement factor H. AB - Complement factor H (fH) is an important regulator of complement activation. It contributes to protection of cells against homologous complement attack. In this study we tested the effect of fH-depletion of normal human serum (NHS) on lysis of antibody-coated sheep and human erythrocytes (EshA and EhuA). In the absence of fH, lysis of sensitised Esh and Ehu was clearly increased. Addition of fH to fH-depleted serum re-established protection of cells against complement similar to that seen with NHS. A fH-derived peptide (pepAred), covering the N-terminal half of SCR 13 in fH, was able to enhance complement-mediated lysis of EhuA significantly. However, the oxidised form of this peptide (pepAox) had no effect. Biotinylated pepAred, but not pepAox, was able to directly bind to cells. Additionally, pepAred competed with direct fH-cell interaction which was observable only after treatment of purified fH with mercaptoethanol. Only pepAred increased the amount of C3 fragments and reduced levels of fH detectable on cells as shown by FACS analysis and radio-immuno assay. Furthermore, fH and factor I (fI)-mediated cleavage of agarose bound C3b into iC3b was decreased in the presence of pepAred. These data indicate that a fH-derived peptide can enhance complement-mediated lysis. We will continue to investigate whether the use of a fH peptide can be exploited for therapeutical purposes. PMID- 11402502 TI - A novel anti-CD18 mAb recognizes an activation-related epitope and induces a high affinity conformation in leukocyte integrins. AB - Monoclonal antibody MEM-148 was previously shown to recognize CD18 chains in a free form unassociated within leukocyte integrin heterodimers, but yet it is paradoxically able to induce a high-affinity conformation in the native, cell surface expressed LFA-1 molecules. Our results based on kinetics of binding, immunoprecipitation and cell-aggregation experiments demonstrate that the mAb does bind to and stabilizes a specific conformation of LFA-1 heterodimers apparently distinguished by an increased affinity to its cellular ligand(s). A similar high-affinity conformation of LFA-1, in which the MEM-148 epitope becomes exposed, is induced also by a Mg2+/EDTA or low pH (5.5-6.5) treatments which may mimic physiologically relevant situations in normal or inflamed tissues. Thus, mAb MEM-148 is a novel valuable tool for detection and induction of specific conformations of human leukocyte integrins. PMID- 11402503 TI - The relationship between HA-1 compatibility and the activation of helper T lymphocyte precursors. AB - The relationship between the compatibility in minor histocompatibility HA-1 antigen and the activation of helper (IL-2 producing) T lymphocyte precursors in vitro was studied in the group of 17 HLA-A2 positive HLA identical siblings. Although the number of pairs studied is still small, no correlation has been found between HA-1 compatibility and helper T lymphocyte precursors activation. The results presented here could suggest the possibility that the HTLp assay does not have to be a relevant parameter for the detection of HA-1 mismatches in HLA identical siblings. PMID- 11402504 TI - Paroxysmal bi-parietal activity induced by full field stroboscopic photic stimulation during sleep. AB - Paroxysmal bi-parietal (vertex) discharges are evoked during sleep by full field stroboscopic light stimulation in approximately 20 per cent of children with a diagnosis of various seizure phenomena. PMID- 11402505 TI - Mechanomyographic responses to continuous, constant power output cycle ergometry. AB - PURPOSE: The purpose of the present investigation was to examine the mechanomyographic (MMG) and electromyographic (EMG) responses to continuous, constant power output cycle ergometer workbouts. METHODS: Eight adult male volunteers (mean age +/- SD = 22 +/- 2 yrs) performed three continuous, one-hour workbouts at 28, 35, and 42% of peak power (Ppeak). RESULTS: The slope coefficients for the mean normalized MMG amplitude versus time relationships were significantly (p < 0.05) less than zero, while the slope coefficients for the mean normalized EMG amplitude versus time relationships were significantly (p < 0.05) greater than zero. CONCLUSION: The results indicated dissociation between the patterns of the mechanical (MMG) and electrical (EMG) activity of the vastus lateralis during continuous cycle ergometry at low power outputs. The increases in EMG amplitude were likely due to the recruitment of additional motor units. The decreases in MMG amplitude across time may have been due to the phenomenon known as "muscular wisdom" and/or decreases in muscular compliance. PMID- 11402506 TI - Electrophysiologic correlates of weakness in L5/S1 radiculopathy. AB - INTRODUCTION: Management of patients with radiculopathy involves estimating the degree of physiologic and anatomic injury, and weighing that to predict the likely clinical course. OBJECTIVE: To determine whether low distal peroneal and tibial CMAP amplitudes correlate with weakness and fibrillations of functionally relevant muscles in L5/S1 radiculopathy (LSR). METHODS: We reviewed clinical and electrophysiologic data in 66 consecutive patients with LSR. RESULTS: A significantly greater number of patients with low peroneal CMAP amplitudes had weakness of L5 (p = 0.025) and S1 innervated leg muscles (p < 0.001). Low tibial CMAP amplitudes were also associated with weakness of S1 innervated muscles (p < 0.038). The association of low peroneal CMAP amplitudes with weakness persisted when weakness of at least 3 muscles was considered in the analysis for L5 (p < 0.0001) and S1 (p = 0.014) innervated muscles. CONCLUSIONS: Low peroneal and tibial CMAP amplitudes may serve as surrogate measures for segmental weakness of functionally relevant muscles in LSR. PMID- 11402507 TI - Comparison of the shock artifacts induced by tripolar and bipolar electrical stimulation techniques. AB - Tripolar and bipolar electrical stimulation procedures were performed on the upper limbs of eight subjects. The mid-forearm was stimulated electrically (tripolar or bipolar) by surface electrodes, and the induced stimulus shock artifacts were recorded simultaneously from the wrist and elbow. During tripolar stimulation, two types of stimulating configurations were utilized: with the center electrode designated as the cathode and the two outermost electrodes connected to a common anode, and vice versa. During bipolar stimulation, the center electrode served as one pole of the stimulator, and one of the two outermost electrodes of the tripolar stimulator was disconnected. The stimulus intensity was kept constant in all stimulating procedures. Artifacts were reduced significantly during tripolar compared to bipolar stimulation, if the outermost electrodes of the tripolar stimulator (which were facing the recording electrodes) were also oriented toward the recording sites during bipolar stimulation and had the same stimulus polarity. Artifacts were slightly reduced in amplitude from tripolar stimulation, if the center electrode were oriented toward the recording sites during bipolar stimulation and had the same stimulus polarity as previously used during tripolar stimulation. PMID- 11402508 TI - Imaginary motor movement EEG classification by Accumulative-Autocorrelation Pulse. AB - Analysis of motor imaginary electroencephalogram (EEG) signals provide a feasible low-level communication channel for handicap people. We propose a classification method for imaginary right and left motor EEG using the Accumulative Autocorrelation-Pulse (AAP) technique. This technique is based on the spatio temporal pulse patterns generated from the accumulative autocorrelation values of selected electrodes in the ongoing EEG data. A feed forward neural network trained with the back propagation learning algorithm is used for classification. The network structure preserves and extracts the pulse-temporal feature patterns of the signal. Classification results reach 100% generalization accuracy in some single subjects and a 91% generalization over all subjects when the correct pair of electrodes are selected. Robust generalization results indicate that the autocorrelation nature of the human EEG signal contains typical patterns for classification in imaginary left and right motor events. PMID- 11402509 TI - The influence of eccentric contractions and stretch on alpha motoneuron excitability in normal subjects and subjects with spasticity. AB - Application of eccentric contractions and muscle stretch are clinically effective in reducing spasticity and increasing ROM (7). This may be explained by a change in the excitability of motoneurons supplying the spastic muscle. Excitability of motoneurons can be indirectly assessed using the H-reflex. Experiments were performed on 20 normal subjects and 17 subjects with spasticity resulting from neurological disorder. Subjects were seated in a secure position and the ankle joint was moved from 30 degrees plantarflexion to 20 degrees dorsiflexion at a velocity of 30 degrees/sec. Sixty eccentric contractions of the triceps surae muscle were performed using a Kin-Com dynamometer (Chattanooga Corp, Tennessee). Two protocols were used: (1) eccentric contractions only, and (2) eccentric contractions with a 5s stretch of the relaxed triceps surae after each contraction. Two sets of 10 H-reflexes were collected from the soleus muscle before (trial 1 & 2) and after (trial 3 & 4) eccentric and eccentric + stretch protocols. The mean peak to peak H-reflex amplitude was calculated for each trial and compared using ANOVA. Eccentric contractions resulted in a significant and maintained increase in the H-reflex in neurological compared to normal subjects (P < 0.05). Eccentric contractions in subjects with spasticity resulted in an increase in motoneuron excitability which may assist in corticospinal activation of motoneurons during voluntary movement. The eccentric + stretch protocol, resulted in a decrease in the mean amplitude of H-reflexes in neurological subjects, however, this was not significant. The application of a stretch following eccentric contractions decreased motoneuron excitability and may thus be beneficial to decrease spasticity whilst strengthening muscle. PMID- 11402510 TI - Electromyographic validation of the trapezius and serratus anterior muscles in military press exercises with open grip. AB - It was studied the trapezius muscle and serratus anterior muscle in 24 male volunteers using a 2-channel TECA TE 4 electromyograph and Hewlett Packard surface electrodes, during the execution of four different modalities of military press exercises with open grip. The results showed that TS acted significantly in the modalities standing and sitting press behind neck, while SI acted in all the modalities, i.e., standing and sitting press behind neck and forward, justifying their inclusion as basic exercises for physical conditioning programmes. PMID- 11402511 TI - Subclinical peripheral neuropathy associated with chronic obstructive pulmonary disease. AB - Peripheral nervous system disorders in COPD (chronic obstructive pulmonary disease) have been found more frequently than usual neurologic practice. We planned this prospective clinical and electrophysiological study to determine the incidence and characteristics of neuropathy in patients with COPD. We studied 49 patients with COPD in whom other causes of polyneuropathy had been excluded. COPD patients were divided into two groups: 21 hypoxemic and 28 normoxemic. Age and sex matched, nonsmoker, 21 healthy subjects were included as the control group. We investigated the results of clinical (neurological symptom score-NSS, neurological disability score-NDS and vibration perception thresholds- VPT) and neurophysiological evaluations in COPD patients and the control group. A value over the mean +/- 2.5 SD of control group were accepted pathologic. NSS results were pathologic in 34% of COPD patients, NDS in 42% and VPT in 94%. Carpal tunnel syndrome was found in 24% of the patients, neuropathy in 55%, and polyneuropathy in 44.8%. In conclusion, the incidence of neuropathy was more than expected, the rate of axonal neuropathy was significantly higher in the hypoxemic group than normoxemic group and the severity of neuropathy was correlated with the degree of hypoxemia. PMID- 11402512 TI - [Economy class syndrome: myth or reality and what prevention?]. PMID- 11402513 TI - [Myelodysplasias and systemic diseases. A non-fortuitous association]. AB - PURPOSE: Myelodysplastic syndromes are clonal hematologic disorders, expanded from myeloid stem cells. A primitive immunologic disorder is discussed. This hypothesis could explain a non-casual association with systemic diseases. The aim of our study is to test this hypothesis. METHODS: We retrospectively investigated the data of 60 patients with myelodysplastic syndromes (group I) hospitalized in our unit from 1990 to 1999. The frequency of systemic disorders was screened and compared to controls (group II). Group II consisted of 120 patients matched for age and sex and hospitalized in the same hospital during the same period. RESULTS: Sixty patients were included (mean age: 83 years old). Myelodysplastic syndrome subtypes were refractory anemia with excessive blasts (52%), refractory anemia (43%) and sideroblastic anemia (5%). Fourteen cases of systemic manifestations were reported in group I (23%) and five in the controls (4%) (P < 0.0001). Systemic manifestations in group I included vasculitis in six cases (42%), polyarthritis in three cases (21%), systemic amyloidosis AA in two cases (14%), relapsing polychondritis in one case, pyoderma gangrenosum in one case and celiac disease associated with a systemic granulomatosis in one case. In the controls, vasculitis was present in four cases and polyarthritis in one. Median age at onset of myelodysplastic syndrome was not influenced by the association with systemic disorders which, in return, have not influenced the myelodysplastic syndromes' subtypes. Myelodysplastic syndromes succeeded to systemic manifestations in 71.4% of cases and could not be attributed to immunosuppressive therapy. CONCLUSIONS: The association of myelodysplastic syndromes with systemic manifestations seems not to be casual. It raises the hypothesis of a primitive immunological disorder in both diseases. Moreover, the description of two cases of systemic amyloidosis and one case of pyoderma gangrenosum might suggest an additional disorder of macrophages or granular cells. PMID- 11402514 TI - [Evaluation of a hospitalization unit opened during the winter influenza epidemic of 1999-2000]. AB - PURPOSE: Last winter, a great many patients with influenza-like infection were admitted to our hospital, leading us to open a specific unit for 6 weeks. We report the evaluation of medical care given to these patients. METHODS: Useful data for evaluating care to patients presenting respiratory infections were determined beforehand by the retrospective analysis of patients' charts. RESULTS: Fifty-seven out of 185 admitted patients (31%) had infectious respiratory symptoms. The mean age was 81 years. Six cases of influenza virus infection, 43 cases of viral bronchitis, six cases of bacterial pneumonia, one superinfected asthma and one septic shock were diagnosed. All patients presented with cardiac and/or chronic pulmonary diseases. Influenza vaccination had been performed in 28 patients (49%). Before hospitalisation, 30 patients (52%) had received antibiotics, and 17 (30%) a steroid therapy. In contrast, only 12 patients (21%) have received anti-infective agents during the hospitalisation. Twenty-five patients were able to go back home and a nursing home was required for 27 patients (47%); five patients died. Tools for improving this specific department in a public hospital are discussed. CONCLUSION: Vaccinations in the elderly appear to be poorly utilized; meanwhile, antibiotic treatments, as well as steroid therapy, are overused. Managing epidemic infections requires attention from the public hospital system. PMID- 11402515 TI - [Sickle cell disease in adults: which emergency care by the internists?]. AB - INTRODUCTION: Sickle cell disease is an inherited disease characterized by the presence of an abnormal haemoglobin. It is the most prevalent genetic disease at birth in the Ile-de-France area. Internists are involved in the management of acute complications, particularly acute vaso-occlusive crisis. CURRENT KNOWLEDGE AND KEY POINTS: Sickle cell disease can be complicated by acute vaso-occlusive crisis, chronic visceral involvement related to the ischaemic process, and infectious complications. In adults, acute vaso-occlusive crisis is the major clinical problem prompting admission to the hospital and the main cause of death. It mainly manifests by osteoarticular pain but other clinical complications can be observed such as acute chest syndrome, priapism, ischaemic or haemorrhagic stroke, abdominal pain and acute multivisceral failure. The treatment of acute vaso-occlusive crisis is symptomatic. Simple transfusion or partial exchange transfusion is required in the more severe form of vaso-occlusive crisis. FUTURE PROSPECTS AND PROJECTS: The management of adult patients with sickle cell disease must be based on a multidisciplinary approach. At the present time, more than 50% of patients survive beyond the fifth decade. This better and longer life in developed countries has resulted from basic investigations and symptomatic treatments. PMID- 11402516 TI - [Large granular lymphocyte proliferations. Clinical and pathogenic aspects]. AB - INTRODUCTION: The clinical course, biological features, and recent data on the pathogenesis of large granular lymphocyte (LGL) leukemia are reviewed. CURRENT KNOWLEDGE AND KEY POINTS: Clonal diseases of LGL disorders can arise from a CD3+, CD57+ T-cell lineage, which are the most frequent, or from a CD3-, CD56+ NK-cell lineage. The diagnosis of LGL leukemia is suspected on the basis of a persistent excess of LGL, usually with neutropenia and splenomegaly. It is assessed by immunophenotypic and molecular studies of T-cell receptor clonality (southern blot, PCR). Association with autoimmune diseases (rheumatoid arthritis, erythroblastopenia, etc.) is a main feature of chronic LGL proliferation. Questions about a viral agent (HTLV1?), facilitation of clonal expansion by cytokines (IL-12, IL-15), and the defective Fas apoptotic pathway are discussed. Treatment of symptomatic LGL proliferations is based on immunosuppressive agents (principally methotrexate and cyclophosphamide). FUTURE PROSPECT AND PROJECTS: The epidemiology, prognosis factors, therapeutics and the pathogenesis of LGL leukemia are unknown. We proposed the creation of a French register of LGL expansions to explore these different aspects. PMID- 11402517 TI - [Hyperparathyroidism induced by lithium. A new case]. AB - INTRODUCTION: Lithium salts, used for the first time in 1949, had proved to be a highly effective preventive measure in bipolar illness. The first report of lithium-induced hyperparathyroidism was suggested by Garfinkel et al. in 1973. About 40 cases have been reported since, suggesting an enhancement of occurrence of hyperparathyroidism in patients cured by lithium carbonate. We report here a new case discovered by a systematic measurement of calcemia after a surgical intervention for a hip joint prosthesis. EXEGESIS: Unusual metabolic features associated with this case of hyperparathyroidism include low urinary calcium excretion, normal cyclic AMP excretion and lack of calcic nephrolithiasis. The mechanism probably results from lithium linking with the calcium receptor on the parathyroid and then stimulating PTH secretion. In the same way it could enhance the tubular reabsorption of urinary calcium. Lithium withdrawal is often inefficient in clinical and laboratory test abnormalities and surgery is usually required. CONCLUSION: It is very important to recognise this particular secondary effect of lithium therapy because clinical symptoms of hypercalcemia can simulate a worsening of the bipolar illness. PMID- 11402518 TI - [Gastroduodenal metastases: an unusual manifestation of lung cancer. Study of two cases and review ot the literature]. AB - INTRODUCTION: Diagnosis of gastroduodenal metastases is rare. Primary tumors are essentially melanomas and breast cancer, and exceptionally lung cancer. EXEGESIS: We report two patients who have a diagnosis of gastroduodenal metastases as initial manifestation of lung cancer. In one case, the patient died 3 weeks after the diagnosis. In the other case, chemotherapy was performed and complete response was obtained for the gastric metastasis. After a few months, node recurrence was diagnosed and the patient died 8 months after the diagnosis. CONCLUSION: We review the endoscopic and non-endoscopic literature and discuss the different histological types and therapeutic strategies concerning these unusual manifestations of lung cancer. PMID- 11402519 TI - [A disseminated form of Langerhans histiocytosis associated with diabetes insipidus and diabetes mellitus]. AB - INTRODUCTION: Langerhans' cell histiocytosis is a rare disorder of unknown etiology characterized by a wide clinical spectrum and varied behavior. Diabetes insipidus is a relatively common feature in Langerhans' cell histiocytosis. The presence of both diabetes insipidus and mellitus associated with histiocytosis in an adult is rare. To our knowledge, only three previous cases have been reported. EXEGESIS: We report the clinical presentation, pathologic findings and clinical progress in an adult female who had disseminated Langerhans' cell histiocytosis (hypothalamic infiltration, multifocal bone involvement) associated with both diabetes insipidus and mellitus. CONCLUSION: The pathogenesis of diabetes mellitus in such an association will be discussed. PMID- 11402520 TI - [Thymoma and disseminated lupus erythematosus. Two new cases and review of the literature]. AB - INTRODUCTION: Thymoma is a tumour originating in the epithelial cells of the thymus, associated with several immunologic disorders. The association of thymoma with systemic lupus erythematosus has rarely been described. We report two cases of this association. EXEGESIS: Description of two cases and a review of the literature. Mr T. was 41 years old when the diagnosis of thymoma and lupus was made. The thymectomy did not influence the evolution of his lupus. Mrs G. had been treated because of a lupus for 8 years prior to developing a thymoma. One year later she presented with erythroblastopenia, which was only sensitive to cyclosporin. CONCLUSION: The association between lupus and thymoma has been reported in 36 cases in the literature. Thymoma is benign in 59% of the cases. The clinical presentation of lupus is nonspecific except for age, median 48 years, and sex ratio, 4:3. The clinical outcome of the lupus is not influenced by the thymectomy. Thymoma may precede lupus with a delay of several years or it may be diagnosed concurrently or several years later. This association is not accidental, though the pathogenic link between these conditions remains unknown. One could suppose that the decrease of the thymic function in the course of thymoma could enhance the expression of autoreactive T lymphocytes as well as the activation of B cells. Patients should be followed after thymectomy because autoimmune diseases, particularly lupus, may develop belatedly. On the other hand, thymoma may be suspected mainly when lupus occurs in patients around 50 years of age. PMID- 11402521 TI - [A neglected tumor. A pleomorphic adenoma of the right parotid gland]. PMID- 11402522 TI - [Horton disease manifesting as periaortitis]. PMID- 11402523 TI - [Hodgkin's disease and acute nephrotic syndrome association]. PMID- 11402524 TI - [Primary cutaneous large cell lymphoma B of the legs and rheumatoid polyarthritis]. PMID- 11402525 TI - [Toxic epidermal necrolysis treated with intravenous immunoglobulins]. PMID- 11402526 TI - Evaluation of certain veterinary drug residues in food. AB - This report presents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of residues of certain veterinary drugs in food and to recommend maximum levels for such residues in food. The first part of the report considers the interpretation of data on inhibition of cholinesterase activity and recommendations arising from an informal meeting on harmonization with the Joint FAO/WHO Meeting on Pesticide Residues. A summary follows of the Committee's evaluations of toxicological and residue data on a variety of veterinary drugs: one anthelminthic agent (ivermectin); four antimicrobial agents (flumequine, lincomycin, oxytetracycline and tilmicosin); six insecticides (cyhalothrin, cypermethrin, alpha-cypermethrin, dicyclanil, permethrin and metrifonate (trichlorfon)); and one production aid (melengestrol acetate). Annexed to the report are a summary of the Committee's recommendations on these drugs, including Acceptable Daily Intakes and Maximum Residue Limits and further information required. PMID- 11402527 TI - [The future of urology]. PMID- 11402528 TI - [The future of urology in Europe: a perspective from the European Urology Association]. AB - INTRODUCTION: In order to be able to influence and monitor future developments for urologists, strategies should be promoted in advance to guarantee the future of the speciality and to accommodate the inevitable changes. Faced with this challenge, the EAU, through its Strategy Planning Office (SPO), has prepared a document which is offered, here, in abbreviated form, to the European and international urological communities for general consideration. MATERIAL AND METHODS: A group of subjects, related to the domains and internal consistency of urology as a speciality, were selected and discussed among the members of the SPO and later submitted to open consultation among distinguished members of the urological community. The topics selected for discussion included: what is urology; urology in the university; sub-specialization in urology; training in urology; does kidney transplantation belong to urology, and others. RESULTS: It is shown that urology is going through an exciting and hazardous transition period. Urology has conflicting problems in its traditional domains due to changes in health care policy, and internal identification problems due to its permanent expansion and sub-specialization options. Weaker points are its relation with primary care medicine (shared care options), the presence and role of urology in institutions such as the university, department of surgery, children's hospitals, administration, etc.; the desegregating effect of the sub specialities; the increasing encroachment of other specialities, and the increasing outpatient effect of technological progress. CONCLUSIONS: An action plan is proposed to confront these changes without losing manpower, internal consistency or social image and improving patient care quality, excellence of training and scientific progress. PMID- 11402529 TI - [Effects of chronic alcohol drinking on rat's prostate. Experimental study]. AB - OBJECTIVE: We have studies both the morphologic and morphometric modifications that the alcohol induces in subjected rats to chronic intake of alcohol and to evaluate the reversibility of this alterations after suppressing the ingesta of alcohol. MATERIAL AND METHODS: The animals underwent diverse outlines of chronic intake of alcohol and diverse morphometric parameters of the prostate were valued at the level of optic microscopy. RESULTS: Two morphologic prostatic patterns was observed according to the studied group of animals (experimental or control). It seems to be that the dose of alcohol was the factor that more it influenced in the morphometric variations of the cells. PMID- 11402530 TI - [Value of a second transurethral resection in the assessment and treatment of patients with bladder tumor]. AB - OBJECTIVE: To analyse the value of a second transurethral resection, repeated within 3 to 6 weeks after the initial resection, in the treatment and the classification of patients with bladder tumour. MATERIAL AND METHODS: We analyse the results of 72 repeated transurethral resection in 23 (32%) patients with T1 G1-2 bladder tumor, 9 (12.5%) with T1 G3 tumor, 31 (43%) with T2a G2-3 tumor and 9 (12.5%) with T2b G2-3 tumor. The evaluated patients are not associated with CIS. RESULTS: Of all 23 T1 G1-2 tumors, 13 (57%) had residual Ta T1 tumor and 1 (4%) T2 tumor. In this group, the second resection changed the treatment in 1 (4%) patient. Of all 9 T1 G3 tumors, 4 (44%) had residual T1 tumor and 1 (11%) T2 tumor. In this group the second resection changed the treatment in 1 (11%) patient. Of all 31 T2a G2-3 tumors, 5 (16%) had residual T1 tumor, 4 (13%) T2 tumor and 6 (19%) T3-T4 tumor. In this group, the second resection changed the treatment in 10 (32%) patient. Of all 9 T2b G2-3 tumors, 2 (22%) had residual T1 tumor, 1 (11%) T2 tumor and 2 (22%) T3-T4 tumor. In this group, the second resection changed the treatment in 3 (33%) patients. CONCLUSIONS: In T1 G1-2 and T1 G3 tumors, a second transurethral resection detect residual tumor in 36% of patients and change the treatment in 6% of patients. In T2a-b tumors, a second transurethral resection detect residual tumor in 50% of patients and change the treatment in 33% of patients. PMID- 11402531 TI - [Preliminary results of the treatment of high grade (T1G3) superficial tumors of the bladder with transurethral resection]. AB - OBJECTIVE: To examine the results of monotherapy with TUR in the treatment of primary T1G3 transitional cell carcinoma (TCC). METHODOLOGY: Thirty-two patients with primary TCC of the bladder were allocated into a surveillance program. Risk factors for progression to muscle-invasive disease were determined. Immediately, projections of disease-free and progression-free survival were calculated. RESULTS: Five patients (15.6%) were lost in follow-up. Twenty-three (85%) had superficial recurrences. Four patients (14.8%) progressed to muscle-invasive or metastatic disease. No independent risk-factors for progression were disclosed. Median disease-free survival was 8 months. Projection of the risk of recurrence at 79 months was 84.9%. Median time to progression has not been reached yet. Projection of progression at 79 months was 46.3%. CONCLUSIONS: The above mentioned treatment schedule is associated with very high recurrence rates. In addition, recurrences are very frequent. Nevertheless, in the medium run, projections of progression suggest that surveillance can be an alternative to other treatments in the management of T1G3 TCC of the bladder. PMID- 11402532 TI - [Role of PSA velocity in the detection of prostate cancer. A study of 986 males]. AB - OBJECTIVE: To analyze the role of PSA velocity (PSAV) in the detection of prostate cancer (Pca) when compared to other valid alternatives. PATIENTS AND METHODS: From a Pca screening program, 986 men were evaluated in two visits (601 of them agreed for a third visit). Serum PSA was performed in every visit (PSA1, PSA2 and PSA3). All Pca diagnosed after PSA1 were excluded. Criteria for biopsy (PSA2 and PSA3) were PSA > 4 ng/ml, or PSAV > 0.7 ng/ml/year. Diagnostic performance of PSAV was compared with other options (PSA alone, DRE, and PSA density -PSAD-). RESULTS: Median age was 57 years. Median interval between visits were 679 days and 852.5 days respectively. During PSA2, 122 biopsies were indicated (91 performed). After PSA3, 78 were indicated and 24 done. This great proportion of not biopsied men was due to refusal. Seven Pca were detected during PSA2, and 5 during PSA3. Sensitivity of PSAV (two draws) was 0.86, specificity 0.95, missed 1 cancer of 7 and needed 7.5 biopsies per cancer. When three PSA samples available, PSAV missed 2 cancers of 5, and 2.7 biopsies per cancer needed. PSA alone detected 86% of the cancers, multiplying by 2 the number of biopsies needed. Not DRE, nor PSAD improved the diagnostic performance of PSAV when combined with this parameter. CONCLUSIONS: Diagnostic performance of PSAV was found to be unacceptable in our hands. The need for a third biopsy in these studies make them difficult to reproduce. Validation of PSAV is a difficult task to achieve, we think its role remains questioned. PMID- 11402533 TI - [Socioclinical and diagnostic characteristics of prostatism in Spain at the end of the 20th century]. AB - RATIONALE: To study sociodemographic characteristics and current diagnostic and therapeutic behaviour with Benign Prostate Hyperplasia (BPH) patients in Spain at the end of the 20th century. METHODS: Sociodemographic, clinical, therapeutic and quality of life (QoL) data from study ESECI-98, conducted in Spain (1998-1999) in patients with BPH. RESULTS: Nine hundred and forty nine patients were evaluable, with a mean (+/- SD) age of 65.6 (+/- 7.8) years, with concomitant diseases (40%), pain/discomfort (34%) and anxiety/depression (23%); QoL score (mean +/- SD) was 72.1 +/- 16.1 on a total possible score of 100. BPH diagnosis was based on symptoms (93%), digital rectal examination (93%) and abdomino-pelvic ultrasonography (76%). PSA was mentioned for the diagnosis of BPH in 54% of the patients and 77% were receiving pharmacological treatment (61% alpha-blockers) during less than 6 months (38%) or more than a year (35%). CONCLUSIONS: In this study the diagnosis of BPH was mainly based on symptoms, digital rectal examination and abdomino-pelvic ultrasonography. It is worth mentioning that PSA for the diagnosis of BPH, was used in a half of the total number of patients. Pharmacological therapy was used in 3 out of every 4 patients. Compared to a previous national study, there is a change on diagnosis and therapy of BPH. PMID- 11402534 TI - [Surgical procedures of the urinary tract and male genital organs. Rates of surgical intervention: variability and its relationship with distance and income]. AB - OBJECTIVE: To valorate the existence of the variability, in surgical rates on urinary tract and masculine genital organs, between the population of Health Centres (HC) adscribed to the Hospital of Ciudad Real. It so, quantify the influence of the distance (from Health Center to Hospital) or the income have in the variability of these rates. MATERIAL AND METHOD: Surgical activity of the period 1996-98. It has been analyzed for every procedure: residence, sex, age and type (coded by CIE 9th MC). The surgical rates, standardized for age and sex, (TFQ) for every HC has been compared whit the mean (t alpha/2 n-1 alpha = 0.99) of the distribution of rates by HC. The relation between the TFQ and distance or income has been analyzed by lineal regression and ANOVA test for one variable, using the Newman Keuls test to assess the difference of the rates between the HC at different distance or income. It has been considered as significative every p < 0.05. RESULTS: The population studied has a TFQ significatively different (p < 0.01). significant relationship exists between the TFQ and the distance. There were no relations between the TFQ for HC and the income. CONCLUSIONS: 42% of the variance (variability) of the distribution of TFQ for HC is due to the distance. PMID- 11402535 TI - [Skin metastasis of long-course prostatic cancer]. AB - OBJECTIVE: To present a case of prostate cancer of long evolution and follow up time, in the one which has been evidenced the appearance of skin metastases at hypogastrium level, inferior extremities root and inguinal zone. METHODS: Patient diagnosed of prostate cancer, treated with radical prostatectomy and followed in conferences during ten years by the neoplasica disease. RESULTS: Appearance of skin metastases of a prostate cancer, after ten years of a radical surgery, in the one which the pathological anatomy demonstrated the local infiltration at seminal bladders level. CONCLUSIONS: The skin metastases of a prostate cancer are extremely uncommon, appearing in most of the occasions in process of very long evolution and in those which the disease has not been able be controlled. PMID- 11402536 TI - [Leiomyoma of the bladder. Report of a case]. AB - Benign bladder tumors of mesenchymal origin have a very low appearance incidence, being the most frequent the leiomyoma. We report new case, making a discussion about the complementary tests needed for the diagnose. PMID- 11402537 TI - [Pelvic solid masses of uncertain etiology. A clinical case]. AB - We report the case of a 73 year old male with a bulky endovesical mass infiltrating near organs at the radiologic studies and urinary obstruction. We specify the diagnostic tests which end in the diagnosis of giant benign prostatic hyperplasia. Regarding the case, we consider the causes of solic pelvic masses and the diagnostic methods available, making a stand on the paramount importance of the hystologic exam. PMID- 11402538 TI - [Juvenile gangrenous vasculitis of the scrotum. Report of a new case and review of the literature]. AB - Juvenile gangrenous vasculitis of the scrotum is a peculiar form of scrotal gangrene of undetermined etiology, with clinical and pathological features of its own, described in 1973 by Pinol et al. We report a new case, considering this entity as part of the differential diagnosis of scrotal gangrene, and a review of relative literature. PMID- 11402539 TI - [Scrotal leiomyoma]. AB - A case of a 32-year-old patient with a diagnosis of leiomyoma of the scrotum was studied. Histologic analyses did not yield evidence of malignancy. Full excision of tumor was achieved and the postoperative period was uneventful. PMID- 11402540 TI - [Benign presacral schwannoma producing obstructive uropathy. Clinical case and review of the literature]. AB - We report a case of a presacral benign schwannoma which causes right hydroureteronephrosis without other clinical findings. After a fine-needle aspiration (FNA) biopsy under computed tomography (CT) guidance, a fusocellular tumour without athypia was demonstrated. With the initial diagnosis of benign schwannoma the patient was operated on, removing the tumour, and with the immunohistochemical examination (reactivity for S-100) this diagnosis was confirmed. PMID- 11402541 TI - [Stevens-Johnson syndrome, unusual urologic involvement]. AB - Stevens-Johnson Syndrome is an adverse drug reaction, considered a severe type of erythema exsudativum multiforme, fatal in about 5%. Characterized by erythema with vesiculobullous lesions in skin and mucous membranes. Implicated drugs are mainly anticonvulsivants and sulphonamides. Prompt recognition and withdrawal of the suspected drug is essential in treatment. Denuded skin areas must be treated with the same principles of burns. Systemic immunosuppression, mainly corticosteroids, remain controversial. We report a case of Stevens-Johnson Syndrome in a 43 year-old male who developed disuria and genital lesions with a detachment of foreskin and glans penis. PMID- 11402542 TI - [Prostatic blue nevus. Terminology standardization of prostatic pigmented lesions]. AB - We expose a new case of prostatic blue nevus. The existing confusion in the terminology of the prostate pigmentarias injuries in reviewed literature is commented and we concluded, based on the opinion of most of authors, with a possible unification of creteries the same ones. PMID- 11402543 TI - [On "The first 20 years of history of Actas Urologicas Espanolas (1977-1996)"]. PMID- 11402544 TI - [Genetic consequences of irradiation of one or both parents (results of experiments on Wistar rats) Exitus lethalis in Wistar rat's progeny after irradiation of one or both parents]. AB - Examination of 2563 offsprings of Wistar rats after irradiation of one or both parents with doses of 0.25, 0.5, 1, 2, 3 and 4 Gy was carried out; the manifestation of lethal effects in the progeny of the first generation in ontogenesis was studied. The level of embryonic death was the highest after irradiation of germ cells of parents at stages of spermatids, spermatozoids and matured oocytes. Following irradiation of both parents with doses of 0.25, 0.5 and 1 Gy at these stages of gametogenesis and 4 Gy at the stage of spermatids and matured oocytes there was a trend of increasing radiation effects caused by the participation of two irradiated germ cells. After irradiation of both parents with doses of 2, 3 and 4 Gy the embryonic death F1 was essentially the same as rates for irradiated females and non-irradiated males. The F1 death rate in early postnatal development exceeded the control only after irradiation with doses of 2, 3 and 4 Gy. The increase in radiation effects in the F1 due to the mating of two irradiated parents appears to be associated with a mechanism demonstrating additivity or synergism. PMID- 11402545 TI - [Effect of mitochondrial energetic state on radiation-induced DNA internucleosomal fragmentation in irradiated thymocytes]. AB - One of the earliest features of apoptosis is a decrease of mitochondrial membrane potential. Here we show that when apoptosis is induced in thymocytes by ionizing radiation, inhibitors of mitochondrial energy production suppress DNA internucleosomal fragmentation. The suppression of fragmentation by inhibitors does not correlate with their effect on mitochondrial membrane potential, as it was observed when membrane potential was decreased (in the presence of inhibitors of respiratory chain, uncouplers of oxidative phosphorylation) or non affected (in the presence of oligomycin, inhibitor of mitochondrial ATPase). PMID- 11402546 TI - [Analysis of DNA status in bone marrow cells of mice internally irradiated with 90Sr ]. AB - The response of bone marrow cells of CBA mice injected with 22.2, 222 and 592 kBq/animal to additional gamma-irradiation (3 Gy) for testing purposes was evaluated using SCG (Comet assay). A decrease in induction of DNA damage right after additional gamma-irradiation was determined. It correlated with bone marrow cell quantity and the tail length before additional gamma-irradiation. The results support the suggestion about the activation of DNA repair in bone marrow cells under exposure to 90Sr in vivo. PMID- 11402547 TI - [The protective effect of thermal treatment before irradiation on CFUs from bone marrow of mice: a potential involvement of heat shock proteins]. AB - It was studied on mice how prior whole body hyperthemia affects a colony-forming ability of bone marrow after gamma-irradiation. It was found that heating of the animals (42 degrees C, 10 min) 18-22 h before their total irradiation (4 Gy) increases 2-fold the level of CFUs8 and CFUs12 determined in the spleen exotest. The induced radioresistance correlated with accumulation of heat shock proteins, HSP70 and HSP25, in tissues of preheated mice. Injection of quercetin (a selective inhibitor of the heat shock protein synthesis) 0.5 h before the heating fully abolished both the subsequent heat shock protein accumulation and the rise in CFUs populations as compared with control. It is suggested that heat shock proteins, whose expression increases in response to hyperthermia, can play a role of endogenous radioprotectors. Possible mechanisms of their protective action under irradiation are discussed. PMID- 11402548 TI - [Effect of prolonged dietary zeolites on survival rate and intestinal reaction toward irradiation in mice of different age]. AB - In model experiments on mice the influence of prolonged dietary consumption of zeolites on a survival rate, an average lifetime and a status of adherent mucous layer of a digestive tube was investigated. It was found that zeolite increased a survival rate and average lifetime in 55- and 95-day old mice exposed to a dose of 4 Gy. Zeolite shivirtuin caused more expressed effect, than pegasin. Apparently, shivirtuin causes the more expressed radioprotective effect, because it does not hinder in the development of compensatory responses in a structurally functional status of adherent mucous layer of an intestine after the exposure. PMID- 11402549 TI - [A possibility to predict the severity of individual damage following the exposure to supralethal radiation dosage. Prediction by early radiation response ]. AB - The experiments on dogs and monkeys have shown that the severity of radiation sickness following the exposure to supralethal radiation doses can be predicted by early response of an individual. For such prediction, the most informative criteria include general severity of clinical manifestations during primary radiation response and evaluated within a few post-damage hours, the degree of tension in cardiac rhythm control, the cortisol level, the endocrine status index. PMID- 11402550 TI - [A possibility to predict the severity of individual damage following the exposure to supralethal radiation dosage. Prediction of working capacity of irradiated rats and dogs by response to pre-exposure physical load]. AB - The experiments on rats and dogs exposed to radiation doses which cause intestinal and cerebral forms of radiation sickness have shown that, within the first 24 hours post-exposure, efficiency impairment can be predicted by changes in some indices of the cardiovascular and endocrine systems after physical load used as a non-radiation testing agent. The goodness of fit of experimental data to the expected result is up to 50-70%, still rising if a number of indices for different sections of the neuroendocrine system are used at a time. PMID- 11402551 TI - [A possibility to predict the severity of individual damage following the exposure to supralethal radiation dosage. Prediction of the clinical manifestation of radiation injury in laboratory animals by response to pre exposure hypobaric hypoxia]. AB - The experiments on rats, dogs and monkeys exposed to radiation doses which cause intestinal and cerebral forms of radiation sickness have shown that the severity of a clinical state can be predicted by cardiovascular and endocrine changes when a pre-exposure test with moderate hypobaric hypoxia is performed. The goodness of fit of experimental data to the exposed result is up to 60-80%, still rising if a set of indices of the neuroendocrine status is used. PMID- 11402552 TI - [A possibility to predict the severity of individual damage following the exposure to supralethal radiation dosage. Prediction of working capacity of irradiated rats and dogs by response to pre-exposure hypobaric load]. AB - In experiments on rats and dogs exposed to radiation doses which cause intestinal and cerebral forms of radiation sickness, hypobaric hypoxia used as an unconventional test of animals' reactivity has enabled the relationship between reactivity and radiosensitivity to be established. Higher post-radiation efficiency is observed in animals with less pronounced changes during a barotest. The goodness of fit of experimental data to the expected result is up to 70%, rising if a set parameters for one index or the sum total of indices are used. PMID- 11402553 TI - [UV-sensitivity of hemoglobin dimers in free state and in valency hybrids: modification by serotonin]]. AB - Changes of oxygen-binding activity of hemoglobin dimers modified by the therapeutical doses of UV-light and serotonin in free state and in valency hybrids are analyzed. The prior role of photodissociation to dimers at the UV radiation action on heme-protein molecules has been shown. It has been observed that the complex between hemoglobin and serotonin is formed in fields of alpha beta-dimers contacts. PMID- 11402554 TI - [Personal computer: physical factors, effect on the user]. AB - The characteristic of the physical factors of a personal computer at user's working place was presented. The bioeffects of these factors were considered and the influence of these factors on health was estimated. PMID- 11402555 TI - [The delayed effects of modulated and non-modulated electromagnetic field on epileptiformic activity in rats]. AB - The modifying influence of the weak electromagnetic field on the development of the audiogenic spasmodic activity in rats was shown. The decrease of lifetime of experimental rats exposed to electromagnetic fields with different parameters and development of tumours (in one set of experiments) was found. PMID- 11402556 TI - [Dynamics of the factors of 137Cs transfer to natural and sown grasses in the long term after the accident at the Chernobyl power plant]. AB - The regularities of changes in factors of 137Cs transfer to natural and sown grasses in the longterm period after the Chernobyl accident are described. The dynamics of changes in factors of 137Cs transfer to sown grasses over the period of 1993 to 1999 was of a non-uniform character. On average, these transfer factors (TF) tended to decrease in that period. A significant tendency for an increase in TF to sown grasses was noted for 1997-1999. The half-life periods of decrease in TF to natural grasses in the period 1993-1999 were shown to be ranged from 4.6 to 24.6 years. It was demonstrated that the differences in the TF dynamics between sown and natural grasses were related to the peculiarities of their development and application of countermeasures. PMID- 11402557 TI - [The simulation of 137Cs distribution in forest ecosystems and prediction of its accumulation by forest products]. AB - A mathematical model of 137Cs migration in forest ecosystem is presented, which describes the behaviour of this radionuclide in the forest litter-soil system, trees, understory and forest animals. The model's parameters for different types of forest ecosystems are estimated and model's adequacy is tested through the use of independent experimental data. The sensitivity of the model's output variables is analyzed to variations in the most significant parameters. The differences in the seasonal and mean annual dynamics of 137Cs concentration in muscles of roe deers and mooses are shown to be defined by specific features of the diets of these animals and variations in 137Cs content in the main diet components. PMID- 11402558 TI - [Simulation of 137Cs accumulation in plants and fungi from soil with irregular vertical characteristics and distribution of radionuclide]. AB - A method of calculation of radiocaesium concentration ratio in plants and mushrooms as a function of radionuclide and soil properties distribution within the root (myzelium) zone is presented. The method allows to predict the long-term dynamics of radiocaesium content in field and forest plants and mushrooms. PMID- 11402559 TI - [90Sr and 137Cs in higher aquatic plants of the Chernobyl nuclear plant exlusion zone]]. AB - The content of radionuclides 90Sr and 137Cs in higher aquatic plants of water objects within Chernobyl NPP exclusion zone has been analysed. Biodiversity of phytocenose was studied and species-indicators of radioactive contamination were revealed. The seasonal dynamics of radionuclide content in macrophytes was studied and the role of main aquatic plant clumps in processes of 137Cs and 90Sr distribution in abiotic component of biohydrocenose was demonstrated. PMID- 11402560 TI - [Laboratory features of acute myelogenous leukemia in elderly patients]. AB - The hematologic features of 103 AML patients older than 60 years(elderly patients) were analysed. Prior to AML a myelodysplastic state was noted in 28% of those patients, 39.3% of patients had a white blood cell count below 3000/microliter, and in 25%, no blast cells were detected in the peripheral blood. Hypoplastic marrows were noted in 25.2%. In regard to the subtypes, M3 was noted less frequently in the elderly patients than in the younger patients. CD34 in blast cells was observed in 53% of patients and dysplastic changes in bone marrow cells other than blast cells were observed in 46% of elderly patients. Chromosomal abnormalities with good and poor prognosis were noted in 3.1% and 25.0% of patients respectively, and the most commonsites for chromosomal abnormalities were chromosomes 5 and 7. MDR1 abnormality was detected more frequently in the elderly patients(68%) than in younger AML patients(38%). We speculated that the laboratory features of the elderly AML patients may be related to the aging of bone marrow and the effects of environmental factors. PMID- 11402561 TI - [Plasma lipid level in the elderly]. AB - We report here the plasma lipid levels and the incidence of hyperlipidemia in the elderly. The prevalence of hyperlipidemia was 20 to 40% and almost the same in the elderly compared to that in young adults. The most important role of hyperlipidemia is that it is a risk factor for atherosclerosis in the elderly as well as in young adults. However, the level at which treatment is started and treatment goals are different between young adults and elderly patients because the incidence of other diseases and complications and the length of life remaining are different. We also describe the management of hyperlipidemia in the elderly. PMID- 11402562 TI - [Technological innovations for the 21st century: synthesis of optical and biotechnologies]. AB - As the population ages in many developing countries, preventive-rather treatment based medicine would be the key to improved and cost-effective health care in the 21st century. To accomplish this goal, we have developed a fully automated DNA assay system, called "optical rapid assay(ORA)" utilizing and combining micromachine and optical technologies. This system consists of innovative preparatory modules, a free flow electrophoresis for separation of DNA from other contaminants in the specimens, an extremely compact and effective microreactor for PCR, and two reflection null comparative ellipsometry for direct detection of hybridized target DNA without any labelled reagents. The total system will undoubtedly contributes to early diagnosis of serious infections such as viral hepatitis, and future standardization of DNA assays. Simultaneously, each module of the system provides a potential tool for R & D of DNA based diagnoses and therapies. PMID- 11402563 TI - [The factor of spread of POCT in USA]. AB - In the U.S., with the adaptation of the Diagnostic Related Group/Prospective Payment System(DRS/PPS) of Medicare, and the spread of the Managed Care such as the Health Maintenance organization(HMO), medical services have become limited, and remuneration of medical practice has been markedly restricted. As a result, hospital management has been pressed, and cooperation, merger, and absorption of hospitals is observed among general enterprises. Hospitals have been making efforts to reduce expenses for survival. I introduced a part of the influence of such revisions on the clinical examination field, and the factor of the spread of point of care testing(POCT). PMID- 11402564 TI - [Present state and future of the self-test in preventive medicine]. AB - The preventive medicine changes from secondary prevention of early detection and early treatment to primary prevention of the avoidance of pathopoiesis and health Crisis. In present state, there is no a clinical laboratory which should respond to this proposition of the fact with the convention at all. As well as clinical laboratory in the hospital, the clinical laboratory in preventive medicine has also been centralized to the Health screening facilities. The immediacy of The clinical laboratory which can be measured, when it wants to measure it, Is required in order to grasp "bio-information" for avoiding the crisis of the disease. It become important that simultaneously, some relating inspection items are measured a case in which it wants to attempt. It waits for the possibility of a disease state grasp from the self measuring device that it is convenient and that the accuracy is high for health-crisis and pathopoiesis avoidance as a primary prevention in order to rapidly improve the accuracy of the weather forecast by the launch of the meteorological satellite. PMID- 11402565 TI - [Self-monitoring of blood glucose]. AB - As the point of care testing(POCT), devices for self-monitoring blood glucose(SMBG) are most common. The problems in SMBG can be found in the popularity of SMBG itself. The reasons for the problems are three. First is the innovation of technology. As each maker has developed devices based on their own intention, device to device difference is too large, so that the standardization is needed. The second reason is the immediate measurement which, done independently of laboratory technicians, is insufficient in the quality control. The third reason is that although the health insurance covers the cost of SMBG, medical institutions buy devices and give them to patients who have limited right to choose for themselves. It is expected that specialists in laboratory medicine should be positively concerned with the standardization, introduction, and quality control of SMBG devices. PMID- 11402566 TI - [From the perspective of the Japan Association of Clinical Reagents Industries]. AB - Simple testing conducted in the hospital laboratory and the Commercial Lab is not considered "time-consuming testing" but prompt testing that enables obtaining data near the patient. In the field of medical care, the need for POCT(Point-of Care Testing) which can test in prompt and minimally invasive ways has been increasing. In a related move, new techniques are being developing one after another to meet those needs. In the future, we will need to appropriately provide intensive service at the central laboratory and prompt service at the bedside while increasing economic efficiency. Because the 21st century is being promoted as the age of self-responsibility, we need to create systems and an environment for patients to manage their own health. In terms of the reduction of medical service fees, it seems that the method of payment directs forward like DRG definitely. Therefore, cooperation among industry and academia is needed to promote OTC test agents as testing before getting a disease and take an active part enlightening the importance of home testing. Currently, long-term medication is administered for an increasing number of chronic ailments going with the aging population. It seems desirable to develop and popularize easy home tests for monitoring chronic ailment and easy test agents for the checking side effects by drug. PMID- 11402567 TI - [The subject matters concerned with use of simplified analytical systems from the perspective of the Japanese Association of Medical Technologists]. AB - The subject matters concerned with use of so-called simplified analytical systems for the purpose of useful utilizing are mentioned from the perspective of a laboratory technician. 1. The data from simplified analytical systems should to be agreed with those of particular reference methods not to occur the discrepancy of the data from different laboratories. 2. Accuracy of the measured results using simplified analytical systems is hard to be scrutinized thoroughly and correctly with the quality control surveillance procedure on the stored pooled serum or partly-processed blood. 3. It is necessary to present the guide line to follow about the contents of evaluation to guarantee on quality of simplified analytical systems. 4. Maintenance and manual performance of simplified analytical systems have to be standardized by a laboratory technician and a selling agent technician. 5. It calls attention, further that the cost of simplified analytical systems is much expensive compared to that of routine method with liquid reagents. 6. Various substances in human serum, like cytokine, hormone, tumor marker, and vitamin, etc. are also hoped to be measured by simplified analytical systems. PMID- 11402568 TI - [Comparison of various reticulocyte counting methods and the standardization of reticulocyte counting]. AB - The reticulocyte count has been conventionally determined using a visual method. Recently, due to technological advances an automated method has come into general use in laboratories. The precision in terms of reproducibility of automated reticulocyte counters has also made rapid progress compared with the visual method. The measurements of reticulocyte counts showed good correlation among the five automated reticulocyte counters and the visual method although the accuracy of an automated method is problematical since it is not sufficient to find the true value. It remains difficult to standardize reticulocyte counting because automated reticulocyte counting is performed in accordance with the procedure specified by the respective manufactures; thus, an external quality control survey was performed for an accuracy assurance of reticulocyte counts with the participation of many laboratories. In the future, it is necessary to develop biological reference materials equal to the true value or to establish other standardization using new technology. PMID- 11402569 TI - [The measurement of immature reticulocyte fractions and clinical significance]. AB - We evaluated automated analysis for the measurement of immature reticulocyte fraction(IRF) in 222 blood samples. IRF was performed by R-3000 & XE 2100(Sysmex), VEGA RETIC LC141(Horiba), ADVIA 120(Bayer medical), CELL DYN4000(Abbott) or GEN'S(Coulter), and compared with measurements obtained by visual counting. A subfraction of IRF was also determined using different modalities on all analyzers. The values of IRF were stable after blood storage for 48 hours at 4 degrees C and 8 hours at room temperature. A good correlation of IRF among these analyzers was obtained(r = 0.629-0.869), but the normal ranges in the percentage of IRF varied according to analyzers. However, an elevation in the percentage of IRF is the first sign of bone marrow recovery after chemotherapy and the bone marrow transplantation in the majority of leukemic patients. We suggest that IRF counting is useful for the estimation of early phase erythropoiesis in patients who have successful bone marrow engraftment following bone marrow transplantation. PMID- 11402570 TI - [Reticulocyte as indication of the erythroid hematopoiesis: reticulocyte fractions in peripheral blood and bone marrow]. AB - Reticulocyte fractions of peripheral blood and bone marrow were measured for hematological disease in 235 patients using automated fluorescent reticulocyte analysis of Sysmex R-3000. The comparison with R-3000 and the manual method of bone marrow measurement showed an excellent agreement with a correlation coefficient of r = 0.933. In the ratio of reticulocyte fractions of marrow blood and peripheral blood, the marrow blood rate of reticulocytes, HFR, MFR, and LFR was 3.3, 6.2, 1.6, and 0.9 times higher than the peripheral blood rate, respectively. Cases in which the marrow reticulocyte rate was over ten times higher than the peripheral blood rate were observed in MDS and megaloblastic anemia at ratio of 55% and 100%, respectively. These findings suggest ineffective hematopoiesis in bone marrow. Immature reticulocyte fraction(10% or more) showed an excellent agreement with granulocyte(500/microliter or more) as an indication of the engraftment of allotransplantation of bone marrow, in which the analysis of reticulocyte fractions showed a useful indication of engraftment. In cases of death after bone marrow transplantation unlike in survivors, reticulocyte fractions decreased after engraftment. PMID- 11402571 TI - [Validity of reticulocyte count testing in reimbursement practice: a cost effectiveness analysis]. AB - Recently reticulocyte count testing has markedly decreased in Japan. We studied the current status of the reduction of reimbursement for reticulocyte count testing and the cost-effectiveness of the test in an academic hospital. The orders of complete blood counts(CBC) accompanied by reticulocyte count testing in Tokai University Hospital declined from 58.9 to 13.3% during the last decade. Among a total of 15,538 test orders for reticulocyte count in outpatients, full reimbursement was refused in 123(0.79%) during a one year period from April 1999 to March 2000. The cost-effectiveness of reticulocyte count testing was evaluated in 719 new outpatients who visited Tokai University Hospital during this period. A useful result(UR) of testing was defined as that which contributed to a change in a physician's diagnosis- or decision-making, relating to a "tentative initial diagnosis" obtained from patient history and physical examination alone. Cost effectiveness for either CBC alone or in combination with reticulocyte count testing was evaluated by calculating clinical effectiveness(the number of URs yielded by a test per patient), economic efficiency(cost required for a test per UR generated), and the cost-effectiveness values(cost required for additional test(s) per additional UR generated). There was a wide disparity in the clinical effectiveness of reticulocyte count twisting among the hospital departments, ranging from 0.53 UR/patient(Gastroenterology-2) to 0(Oral Surgery, Neurology, Urology). Poor clinical effectiveness shown in particular hospital departments endorsed the increasing percentages in reimbursement rejection by the payer of health insurance. The test should be ordered more carefully in consideration of reimbursement and in regard to its cost-effectiveness among different patients groups. PMID- 11402572 TI - [Procedures for the hemoglobin value assignment to guarantee the traceability against the international hemoglobin standard: a proposed plan]. PMID- 11402573 TI - [An approach for the quality control in clinical microbiology laboratory by antibiotic susceptibility]. AB - It is important to develop a system for the prevention of various errors and the maintenance of the high quality level in the department of laboratory medicine. We developed a computer program to infer the species of microorganisms by assessing the minimal inhibitory concentration of respective microorganisms to various antibiotics, antibiotic susceptibility patterns(ASP). Retrospective analysis of 3,405 strains from 39 species using this program resulted in a sensitivity of 91.4% and specificity of 98.9%. Furthermore, in 1,660 strains isolated from September 1999 to August 2000, the efficacy of this program was evaluated using both this program and conventional identifying method such as VITEK, api and ATB. The discrepancy between this inferring method and conventional method was noted in 179 strains, revealing errors in 12 strains. We conclude that this ASP inferring program is useful for preventing errors and the maintenance of high quality examination by combining with conventional methods in the laboratory. PMID- 11402574 TI - [Clinical laboratory data and reference intervals standardized in Fukuoka]. AB - In Fukuoka whose population is approximately five million inhabitants, surveys on the accuracy of laboratory data have been performed by the Fukuoka Prefecture Medical Association for the last 30 years. We have been attempting to evaluate the data for routine use since 1988, and it has become possible to share laboratory data between all institutions in Fukuoka prefectures. As a result, reference intervals for 23 clinical chemistry analytes were established in 1995, to which were added in 1996 five serum protein constituents that have been utilized for clinical examinations. Methods for documentations and monitorings the data obtained in the prefecture were also established, standardization of the above analytes extended to 97% of the institutions in the prefecture. Results for 14 of the 23 clinical chemistry analytes have become highly reliable and clinically useful as differences between institutions in terms of results have narrowed. Standardization of other analytes is now in progress. PMID- 11402575 TI - Epidemiology of erectile dysfunction. AB - Epidemiologic studies are descriptive and analytical. The prevalence of erectile dysfunction in various community studies has varied from as low of 7% to as high as 52%. The prevalence of erectile dysfunction increases with age. Incidence data are scarce but a recent study of white males in the United States described an incidence of 26 cases per 1000 man-years. Risk factors for erectile dysfunction include certain medications, such as cardiovascular medications and psychotropic drugs, and chronic diseases, particularly neurologic diseases and diabetes mellitus. PMID- 11402576 TI - Integrative erectile biology. The effects of age and disease on gap junctions and ion channels and their potential value to the treatment of erectile dysfunction. AB - Initiation, maintenance, and modulation of corporal smooth muscle tone are critically dependent upon agonist-induced changes in intracellular calcium levels and mobilization as well as transmembrane calcium flux. The transient control of myocyte excitability and contractility at the cellular level is inextricably linked to membrane potential, which, in turn, is modulated by potassium ion efflux through one of the four known corporeal smooth muscle potassium ion channels. Corporal tissue responses are subsequently coordinated by means of the movement of intracellular second messenger molecules (i.e., IP3, cAMP, cGMP) and ions (i.e., K+ and Ca2+) among the corporal myocytes by means of intercellular communication through gap junction channels. Knowledge of the critical contribution of these interlinking cellular (nonjunctional ion channels [e.g., maxi-K]) and tissue (gap junction channels [e.g., connexin 43]) systems to the modulation of erectile capacity has provided the scientific rationale for the promulgation of the successful preclinical testing of hSlo ion channel gene therapy for the normalization of erectile status in both aged and diabetic rats. PMID- 11402577 TI - Pharmacology of erectile function and dysfunction. AB - Central regulation of the erectile process involves several transmitters, including dopamine, serotonin, noradrenaline, and nitric oxide, and peptides, such as oxytocin and ACTH/alpha-MSH. These systems may be targets for future drugs designed to treat erectile dysfunction. Peripherally, the different steps involved in neurotransmission, impulse propagation, and intracellular transduction of neural signals in penile smooth muscles need further investigation. Continued studies of the interactions between different transmitters/modulators may reveal new combination therapies. Increased knowledge of the changes in penile tissues associated with erectile dysfunction may explain the pathogenetic mechanisms and help to prevent the disorder. PMID- 11402578 TI - Physician-patient dialogue and clinical evaluation of erectile dysfunction. AB - Erectile dysfunction affects 31% to 52% of all men. Although considerable advances have been made in the diagnosis and management of erectile dysfunction, the inadequate knowledge of erectile dysfunction by health care providers still precludes them from initiating candid discussions with their patients. This article provides the health care professional with the ability to establish comfortable doctor-patient dialogue and to clinically evaluate erectile dysfunction in a goal-directed manner. The goal of evaluation is to find the appropriate treatment for the individual, dictated primarily by factors like the patients' preferences, comfort, cost, and the availability of treatment modalities. PMID- 11402579 TI - Smooth muscle electromyography. AB - This article addresses smooth muscle physiology, electromechanical and pharmacomechanical coupling, computed-based interpretations, and the role of electromyography in the diagnosis of erectile dysfunction. PMID- 11402580 TI - Psychogenic erectile dysfunction. Classification and management. AB - Psychogenic factors are involved alone or in combination with organic causes in a substantial number of cases of erectile dysfunction. Epidemiologic studies have implicated the role of depressed mood, loss of self-esteem, and other psychosocial stresses in the cause of erectile dysfunction. A new definition and classification of psychogenic erectile dysfunction has been proposed based on recent clinical and research findings. According to this new classification, psychogenic erectile dysfunction is categorized as generalized or situational type, with subcategories of each type proposed. Traditional treatment approaches for psychogenic erectile dysfunction have included anxiety reduction and desensitization procedures, cognitive-behavioral interventions, guided sexual stimulation techniques, and couples' or relationship counseling. Recently, these approaches increasingly have been combined with pharmacologic therapy such as sildenafil. Special situations have been identified in which combining psychosocial interventions with medical therapy is recommended. These situations include problems of sexual initiation, low sexual desire, other sexual dysfunctions, and significant couples' or relationship problems. More research is needed on the role of psychosocial interventions in the treatment of erectile dysfunction. PMID- 11402581 TI - Hormonal erectile dysfunction. Evaluation and management. AB - The clinical diagnosis of hypogonadism in the adult is difficult to establish on the basis of a history and physical examination and universally requires biochemical investigations. A serum testosterone determination is justified in men complaining of erectile dysfunction with or without alterations in sexual desire. Among the causes of erectile dysfunction, hypotestosteronemia rates are low. The prevalence of erectile dysfunction particularly is common at a period in life when alterations occur in male hormonal environment. The treatment of hypogonadal erectile dysfunction, regardless of age, is readily available, safe, and effective. The positive impact of treatment on the overall quality of life can be significant. The presence of erectile dysfunction in an aging man (> 55 years) does not imply the presence of hypogonadism, and, even if the two conditions are present, the indications for treatment require good clinical judgment. Persistent low testosterone levels may have significant detrimental effects in other organ systems; therefore, a timely diagnosis of androgen deficiency and appropriate treatment may have significant effects outside the narrow field of sexual performance. PMID- 11402582 TI - Neurologic erectile dysfunction. AB - Neurologic erectile dysfunction presents a diagnostic and treatment challenge to the internist and urologist. Multiple chronic disease modalities and traumatic etiologies exist. Education regarding these conditions and a detailed and thorough history and office work-up are the best resources for the clinician. Treatment can follow the model of proceeding from the least to most invasive procedure (process of care), taking into account patient and partner satisfaction. Because the psychology of grief and loss may enter into treatment of some neurologic conditions (e.g., erectile dysfunction after radical retropubic prostatectomy, spinal cord injury, or chronic diseases), a whole patient approach encompassing psychotherapy is warranted. PMID- 11402583 TI - Vasculogenic impotence. Arterial and venous surgery. AB - Arterial revascularization and venous ligation procedures have been introduced within the past 2 decades. Each procedure has in common with the other the fact that initial applications of the operations were widespread among the population of men with vasculogenic erectile dysfunction. In each case, disappointing long term results led to more limited use of surgery targeting specific groups that clearly would benefit from the procedures. The wider application of these procedures in vasculogenic erectile dysfunction is not supported by the available results. The Clinical Guidelines Panel of the American Urological Association supported this view in 1996 after a meta-analysis of literature reports and declared that venous and arterial surgery was not justified in routine use, especially in patients with arteriosclerosis. Further studies are likely to refine patient selection but are unlikely to expand the therapeutic use of these procedures. PMID- 11402584 TI - Oral drug therapy for erectile dysfunction. AB - Oral drugs are a well-established, first-line therapy for erectile dysfunction. As a result of the success of sildenafil, a plethora of new drugs for erectile dysfunction are on the horizon. Apomorphine and IC351 are in late phase III development. Vardenafil (Bayer, New Haven, CT), a PDE5 inhibitor, and the combination of yohimbine and L-arginine (NitroMed, Boston, MA) are in early phase III development. Early clinical and preclinical studies are investigating new phosphodiesterase inhibitors, cyclic AMP activators, alpha-adrenergic antagonists, dopamine agonists, melanocyte-stimulating hormone, potassium channel modulators, endothelin antagonists, and new nitric oxide donors. The future is bright for this infant field of sexual pharmacotherapy. PMID- 11402585 TI - Vacuum constriction and external erection devices in erectile dysfunction. AB - The development of external erection devices has evolved as the social stigma surrounding the treatment of erectile dysfunction has gradually disappeared during the late twentieth century. Although the success of surgical and medical therapy for erectile dysfunction has been documented, especially since the introduction of sildenafil citrate, patient demand for an effective, noninvasive, drug-free management of erectile dysfunction has remained. As the population continues to age, acquiring the comorbidities commonly associated with erectile dysfunction, such as hypertension, diabetes mellitus, and atherosclerotic vascular disease, the demand for such treatment should persist. This article examines the development, mechanism of action, efficacy, and patient satisfaction with regard to vacuum constriction and external erection devices. PMID- 11402586 TI - Intracavernosal injection and intraurethral therapy for erectile dysfunction. AB - Although the search for newer and more effective oral therapies is continually expanding, intracavernosal and intraurethral treatments continue to keep pace and are expected to remain in the clinician's armamentarium in the years to come. Unless there are contraindications, oral therapies are effective, have minimal side effects, and are first-line treatment. Some patients who have failed intraurethral and intracavernosal injection therapies previously have been shown to have successful outcomes with oral sildenafil in 56% to 57% of cases. If oral therapies are ineffective or have undesirable side effects, the intraurethral or intracavernosal routes are the next approach that should be prescribed. PMID- 11402587 TI - Penile prosthesis implantation. AB - The development of effective systemic therapy for the treatment of erectile dysfunction has resulted in a significant increase in the number of men presenting for treatment. Not all men with erectile dysfunction will respond to systemic therapy; those who fail may be candidates for penile prosthesis implantation if second and third lines of treatment also fail or are rejected by the patient and his partner. Penile prosthesis implantation continues to play a role in the treatment of erectile dysfunction. There is a potential for the number of penile prosthesis implantation procedures to actually increase. The ideal penile prosthesis is a three-piece inflatable device that permits good penile flaccidity and increases in size and becomes rigid with inflation. PMID- 11402588 TI - Ejaculatory physiology and dysfunction. AB - The sequence of events encompassing ejaculation has been well described. Multiple disease processes can result in ejaculatory dysfunction. Evaluation and subsequent treatment of ejaculatory dysfunction is possible using behavioral, mechanical, and medical and surgical modalities. Further elucidation of ejaculation is now taking place at the molecular level. PMID- 11402589 TI - Peyronie's disease. AB - Peyronie's disease remains one of the most perplexing diseases in urology. With continued basic research in wound healing and scarring, the understanding and management of this frustrating disease will improve. Initial treatment of Peyronie's disease should be conservative, with expectant therapy and medical management. Once the penile curvature and plaque have stabilized, patients with severe deformity can be offered surgery depending on their symptoms and complaints. Patient selection is the key to proper treatment. Less experienced surgeons should limit themselves to medical management or simple surgical management of the disease, including plication or Nesbit procedures. Penile lengthening procedures or the placement of complicated penile prosthesis should be reserved for surgeons with familiarity and expertise in this type of reconstruction. Education on the pathogenesis and natural history of the disease will allow the patient and his partner to make an informed decision in regards to treatment options and expected outcomes. PMID- 11402590 TI - Priapism. From Priapus to the present time. AB - Advances in the pharmacotherapeutic options available to treat erectile dysfunction over the past decade have transformed the field of impotence. The initial foray into this field with intracavernous injections of papaverine rapidly expanded the number of men seeking attention for priapism, a previously rare clinical condition. The recent widespread use and acceptance of oral agents for the treatment of erectile dysfunction, with a reduced incidence of priapism has decreased the number of men at risk for injection-related prolonged erections. The use of recreational drugs (cocaine) and perineal trauma leading to presentations of priapism seem to be rising in incidence. The urologist remains the consultant-specialist ultimately responsible for these men and should be cognizant of the array of treatments described for this condition. Early determination of the state of corporal oxygenation is essential and will define the optimal management approach. A wide range of medical conditions and risk factors may be etiologic and should be elicited from the patient at the initial interview. Low-flow ischemic priapism requires a rapid resolution, often achieved through use of alpha-agonists orally or by direct injection into the penile circulation, whereas nonischemic priapism can be treated more conservatively. Research into this condition has only recently been initiated. Through greater understanding of the pathophysiology of priapism, the clinician may become armed with etiology-specific medical alternatives providing timely detumescence for men with prolonged erections. PMID- 11402591 TI - Female sexual dysfunction. AB - The ideal approach to female sexual dysfunction would be a collaborative effort between therapists and physicians and would include a complete medical and psychosocial evaluation, and inclusion of the partner spouse in the evaluation and treatment process. Despite significant anatomic and embryologic parallels between men and women, the multifaceted nature of female sexual dysfunction clearly is distinct from that of the man. The clinician cannot approach female patients or their sexual function problems in the same fashion as in male patients. The context in which a woman experiences her sexuality is equally if not more important than the physiologic outcome she experiences, and these issues should be determined before beginning medical therapy or determining treatment efficacies. PMID- 11402592 TI - Socioeconomic considerations in erectile dysfunction treatment. AB - The socioeconomics of erectile dysfunction have been changed greatly by the introduction of the successful oral therapy, sildenafil. The population of patients presenting for treatment of erectile dysfunction has expanded significantly and rapidly. This article addresses the descriptive and analytic economic implications of erectile dysfunction treatment. PMID- 11402593 TI - Laparoscopic radical prostatectomy: technique. AB - Ongoing progress in laparoscopy has permitted extension of its indications for performance of complex oncologic and reconstructive surgeries such as radical prostatectomy. Laparoscopic radical prostatectomy is now feasible when performed by urologists trained in laparoscopy. Although the procedure is technically rewarding, it is more important to evaluate it in regards to efficacy. The short term oncologic and functional efficacy rates (regarding urinary continence) seem equivalent to the rates for open surgery. Longer follow-up is needed to assess the preservation of potency with the laparoscopic technique. The integration and application of open nerve-sparing techniques to the laparoscopic approach should be feasible owing to the excellent visual accuracy at the prostatic apex. Initially, the operating time is lengthy (> 7 hours), but the length improves considerably with each series of 20 or more cases. Overall, the learning curve for laparoscopic radical prostatectomy is approximately 50 cases. Whether the laparoscopic approach will ever be as time efficient for the urologist as open radical prostatectomy remains to be seen. The authors' preliminary impression from the patient's perspective is that laparoscopic prostatectomy may offer significant advantages over open surgery in terms of the immediate postoperative quality-of-life concerns. PMID- 11402594 TI - [Serotonin may be the key to the treatment of "irritable bowel disease"]. PMID- 11402595 TI - [High time to test hypothermia in the treatment of stroke]. AB - There is a revived clinical interest in hypothermia as a neuroprotective intervention in brain ischemia. This originates from the experimental finding that hypothermia of only 3 degrees C-4 degrees C below normal body temperature completely abolishes neuronal damage from an ischemic insult that is lethal in normothermia. The idea that hypothermia protects cells from ischemic damage mainly by lowering metabolic rate is challenged. We propose that detrimental protein-lipid interactions in brain cells, that are activated during and following brain ischemia and that mediate adverse cellular signaling and lead to cell death, are blunted or prevented by mild hypothermia. PMID- 11402596 TI - [Drugs affect and are affected by body temperature]. AB - There are several connections between drugs and cold. Drugs can lower body temperature; ambient temperature together with the adaptive capacity of the organism can influence various physiological and also pharmacological processes; furthermore, hypothermia can have an effect on drug pharmacokinetics and pharmacodynamics which is of considerable clinical importance, since the application of hypothermia as a therapeutic adjuvant is on the rise. The present paper discusses different aspects of these problems, concluding with a proposal that it may now be time to ask for the results of a "cold effect test" before a new drug is accepted for common use, especially in hypothermic patients. PMID- 11402597 TI - [Burn injuries in Sweden 1987-1996. The number of hospitalized patients has been reduced and the mortality lowered by 70 per cent]. AB - Continuous improvement in burn trauma care has led to decreasing mortality, morbidity, and length of hospital stay. Current data from the Swedish population is lacking, which was the reason for this investigation. Data was gathered from the Swedish database for medical care (based on ICD-9). All Swedish care providers are enrolled and assumed to report data. Hence, the quality of the database is considered reasonably good, although data from individual patients may be incorrectly recorded due to clerical errors. Available data indicate that the number of burn victims seeking medical care has decreased by 16% during the past decade, whereas mortality has decreased by 70%! The demographic data is otherwise similar to what has previously been reported in the international literature. PMID- 11402598 TI - [Arthrosis among the elderly. Conservative therapy as a first choice]. AB - Osteoarthritis, a collective term for late joint destruction, is one of the most common causes of disability among the elderly. Since the pathogenesis is unknown, there is no causal therapy. Treatment is therefore primarily conservative and focused on pain control. However, among elderly people, overall results after joint replacement are excellent. Moreover, outcome is continually improving, especially thanks to national quality registers. PMID- 11402599 TI - [Primary hyperparathyroidism is common among postmenopausal women. Identification of genetic risk factors can contribute to individualized treatment]. AB - Primary hyperparathyroidism (pHPT) is commonly seen in postmenopausal women. Along with the clinical characterisation of the disease, studies of molecular genetics have contributed to increased understanding of the etiology of pHPT. Genetic association studies have revealed that certain vitamin D receptor polymorphisms relate to the development of sporadic pHPT. A new type of familial pHPT was recently discovered. Studies of parathyroid adenomas have demonstrated that the tumor suppressor gene MEN1 and the oncogene cyclinD1 are of importance for the tumorigenesis. PMID- 11402600 TI - [Silent salpingitis. Does it exist?]. AB - The diagnosis of salpingitis based solely on clinical criteria is inaccurate, with both low specificity and sensitivity. Laparoscopy has therefore become a valuable diagnostic tool in clinical practice and essential in clinical research on salpingitis. Different types of evidence indicate that atypical salpingitis without abdominal pain and discrete or absent symptoms is a common etiology of tubal factor infertility. A low threshold for suspecting salpingitis has been recommended to augment the sensitivity of clinical diagnosis. This leads to lower specificity and thereby a greater number of false positive diagnoses and unnecessary antibiotic treatment. Outpatient biopsy from the endometrium for histopathology and chlamydia testing might augment the specificity in cases with discrete symptoms, and should be investigated further. The sensitivity of laparoscopy is low for endosalpingitis without affection of the serosa, and might be augmented by using minibiopsies and chlamydia PCR from the tubal mucosa. The most significant measure toward reducing the sequelae of salpingitis is the combatting of chlamydia infection through screening programs and qualified partner management. PMID- 11402601 TI - [A case report. Endometriosis caused colonic ileus, ureteral obstruction and hypertension]. AB - Laparoscopy and palpation offer only a rough estimate of the extent of endometriosis. Consequently, endometriosis involving the bowels and urinary tract is under-diagnosed. Bowel obstruction and retroperitoneal endometriosis with obstruction of the ureter are uncommon conditions, but awareness of them is important. Ureteric obstruction develops slowly from periureteral fibrosis and often results in an asymptomatic hydronephrosis, loss of renal function and hypertension. Although renography is the first line of choice in investigation of the upper urinary tract in cases of suspected ureteric obstruction, ultrasound of the kidneys may be useful in the hands of the experienced gynecologist as a screening tool at consultation. The rationale for this recommendation is that ureteric obstruction and hydronephrosis often occur simultaneously. We present a case with bowel obstruction mimicking sigmoid carcinoma, ureteric obstruction and hypertension, caused by endometriosis, where the diagnostic difficulties are illustrated. Collaboration between gynecologist and urologist is essential in selected cases of endometriosis. PMID- 11402602 TI - [Reduced sick leave without formal change of rules]. PMID- 11402603 TI - [Distribution of health care services. Almost the same costs of health care services for women than for men]. PMID- 11402604 TI - [Are only general practitioners lost? Identity crisis of general practitioners- crisis of health care services?]. PMID- 11402605 TI - [Alcoholism--uncurable or curable foolishness?]. PMID- 11402606 TI - [Folic acid and the risk of neural tube defects]. PMID- 11402607 TI - [Debriefing--just in any way?]. PMID- 11402608 TI - [An example of interactions between SSRI preparations and tolterodine?]. PMID- 11402609 TI - [Better information on vaccinations at maternal-child health centers--but surely it takes time!]. PMID- 11402610 TI - [Why did not the county commissioner laugh?]. PMID- 11402611 TI - Full mouth reconstruction in two visits. PMID- 11402612 TI - Time marches on. PMID- 11402613 TI - Clinical and laboratory evaluation of penicillin resistant Streptococcus pneumoniae in relation to the mutations of pbp 1a, pbp2b and pbp2x. AB - A total of 210 isolates of Streptococcus pneumoniae (S. pneumoniae) were selected randomly to examine drug susceptibility which were obtained from out- and in patients who visited Kurume University Hospital and other affiliated hospitals through May 1998 to September 1999. The prevalence of penicillin resistant S. pneumoniae in this study was 39.5% and was compatible with those of previous reports in Japan. From the clinical aspects, the resistant strains of S. pneumoniae have been shown not to be so highly virulent comparing with sensitive strains that only few severe or mortal cases had been reported. Carbapenems, glycopeptides, and fluoroquinolones were shown to be highly active against penicillin-resistant S. pneumoniae strains as evidenced by the low minimum inhibitory concentrations (MICs) though LVFX showed 4 micrograms/ml or higher MICs against some strains. Regarding to the mechanisms of penicillin resistance, penicillin binding proteins coding gene (pbp1a, pbp2x and pbp2b) mutations that cause modifications in these proteins were also examined for all isolates. The multivariate analysis showed statistically significant correlation between higher MIC of penicillin and cephem and pbp1a mutation while no significant contribution of pbp2x and pbp2b to the resistance was demonstrated. Additionally, no significant correlation between pbp mutation and MIC of carbapenem was observed. Furthermore, there were two highly penicillin resistant S. pneumoniae strains with no pbp mutations. Thus the pbp mutations alone were not responsible for the elevation of MIC all beta-lactams. Nevertheless the pbp mutations were detected in advance of actual MIC elevations by inducement experiment in vitro. It indicated that penicillin resistance might be detected earlier than conventional methods. In previous reports some other responsible genes for penicillin resistance were demonstrated. Therefore, it might be possible to presume exact values of MICs of beta-lactams against resistant strains of S. pneumoniae by detecting still unknown genes other than pbps. PMID- 11402614 TI - The significance of bronchoscopy for the diagnosis of Mycobacterium avium complex (MAC) pulmonary disease. AB - To investigate the usefulness of bronchoscopy for the diagnosis of Mycobacterium avium complex (MAC) pulmonary disease, we retrospectively reviewed the clinical charts, and radiographic and bacteriologic findings of all patients who were admitted to our hospital between 1994 and 2000, and who fulfilled the 1997 American Thoracic Society (ATS) criteria for MAC pulmonary infection. A total of 132 patients were diagnosed as affected by MAC pulmonary disease during that period. Of these, bronchoscopic examination was performed in those patients who showed negative sputum smear for mycobacteria on three consecutive days (n = 43) or who could not expectorate sputum (n = 2). Of 42 patients, sputum culture was positive for MAC in 34 patients (81.0%). Bronchial washing sample was smear positive for MAC in 17 of 39 patients (43.6%), and culture-positive for MAC in 33 of the 39 patients (84.6%). Transbronchial lung biopsy (TBLB) specimens revealed specific findings (epithelioid cell granuloma and/or acid-fast bacilli) in 14 of 38 patients (36.9%). Bronchial washing of all patients who showed specific histology in TBLB grew MAC in culture. Based on the bronchoscopic examination, we could diagnose MAC pulmonary disease in 36 patients. In addition, smear and polymerase chain reaction (PCR) results of bronchial washing made possible an early diagnosis of MAC pulmonary disease in 15 patients. We examined the relation of CT findings to bronchial washing results. Isolation of MAC in bronchial washing is significantly related to small nodular opacity around the ectatic bronchi on the CT scan (p = 0.016). In our retrospective study, in sputum smear negative patients with MAC pulmonary disease, MAC isolation by culture of bronchial washing was no more frequent than that with sputum culture. However, bronchial washing is useful to differentiate infection from casual isolation of MAC. In addition, we could make early diagnosis of MAC pulmonary disease based on smear and PCR results of bronchial washing. To make a diagnosis of MAC, bronchial washing is superior to TBLB, and should be done in the bronchus which drains the area revealing small nodular opacity around ectatic bronchi. PMID- 11402615 TI - Evaluation of simultaneous excision of pancreatic cancer and the surrounding blood vessels. AB - Of the 139 patients who underwent excision for invasive cancer in the pancreatic duct at Kurume University Hospital between January 1965 and December 1998, the subjects were 38 patients in whom blood vessels around the cancer were simultaneously excised. The surgical methods were pancreatoduodenectomy (PD) in 31 patients, distal pancreatectomy (DP) in 5, and total pancreatectomy (TP) in 2. The excised blood vessels were the portal vein alone in 32 patients, the artery alone in 1, and both portal vein and artery in 5. Excision of the portal vein was performed by circumcision in 25 patients and by segmentectomy in 12. The range of circumcision was 1.0-7.0 cm (mean, 3.5 +/- 1.4 cm), and the blocking time of the portal vein was 8-36 min (mean, 19.5 +/- 8.8 min). Of the 25 patients who underwent circumcision, reconstruction was performed by end-to-end anastomosis in 23 and by transplantation of the autologous vein between the ends in 2. Of the 12 patients who underwent segmentectomy, direct suture was performed in 10, and transplantation of an autologous vein patch was performed in 2. Postoperative complications occurred in 14 of the 32 patients. In 5 of the 6 patients who underwent excision of the artery, reconstruction was performed by end-to-end anastomosis in 3 and by transplantation of the autologous vein between the ends in 2. Postoperative complications did not occur in the patient who had undergone excision of the artery alone, but 4 of the 5 patients who had undergone simultaneous excision of the portal vein and artery had postoperative complications, of whom 2 died during the period of hospitalization. Three patients with pv0, pv1 or pv2 survived for more than 3 years. Because some of the patients who had undergone excision of the portal vein alone survived for a long time and this method is relatively safe, this surgery can be generally applied, but simultaneous excision of the portal vein and artery should be carefully applied because the incidences of postoperative complications and death during the period of hospitalization are high. With the development of surgical techniques and postoperative control, simultaneous excision of pancreatic cancer and the surrounding blood vessels has become safe, but this method should only be applied to patients who have the potential to recover completely. PMID- 11402616 TI - Efficacy of targeted screening for childhood hypercholesterolemia in Japan. AB - The aim of this study is to evaluate the efficacy of targeted screening on the basis of the results from the universal screening of Japanese children. A total of 698 children consisting of 348 in the 3rd grade of elementary school (third graders) and 350 in the 3rd grade of junior high school (ninth graders) were enrolled in one local school district, during the period 1985 to 1998. When body mass index was used as the sole indicator, the sensitivity in detecting hypercholesterolemia was 14.3% in the case of third graders, and 14.7% in that of ninth graders. Family history was investigated through a questionnaire inquiring about the history of 6 atherosclerosis-related diseases in 6 relatives. The sensitivity on the basis of family history was calculated as 0% to 54.5%. According to the atherosclerotic index calculated from the measured levels of total and HDL cholesterol, sensitivity of abnormal levels of atherosclerotic index in children was almost the same as those of hypercholesterolemia. Using targeted screening, it was possible to detect only around half or less of the subjects with hypercholesterolemia. With respect to the sensitivity of the screening, universal screening is recommended for detecting hypercholesterolemia in Japanese children. PMID- 11402617 TI - Transient heart murmur in the late neonatal period: its origin and relation to the transition from fetal to neonatal circulation. AB - To elucidate the origin of transient heart murmur during the late neonatal period, we examined 50 neonates with this type of heart murmur and compared them with 50 controls. We serially examined the morphology of and blood flow in the main pulmonary artery (MPA), the right pulmonary artery (RPA), and the left pulmonary arteries (LPA) using two-dimensional and Doppler echocardiography. The heart murmurs were first noticed at 6 to 60 days after birth (mean 33 +/- 14). At that time, the diameters of both the RPA and the LPA in the heart murmur group were significantly smaller than those in the control group, and the velocities of blood flow in the right and left pulmonary arteries in the heart murmur group were significantly greater than those in the control group. When the heart murmur disappeared, the diameters and the flow velocities of both the RPA and the LPA were not different compared with the control group. Two cases in the heart murmur group continued to have a heart murmur and were diagnosed as having intrinsic congenital peripheral pulmonary artery stenosis. In conclusion, our findings suggest that a transient heart murmur in the late neonatal period is caused by transient branch pulmonary arteries stenosis during the transitional circulation from fetus to neonates. PMID- 11402618 TI - In vitro transmission of HCMV between fibroblasts and peripheral blood leukocytes in the presence of IL-4. AB - We demonstrated transmission of human cytomegalovirus (HCMV) from the human lung fibroblast MRC-5 to peripheral blood leukocytes (PBLs). mRNA of the HCMV immediately-early (IE) antigen was detected in PBLs cultured with IL-2 or IL-2 + IL-4 that made direct contact with HCMV-infected MRC-5, whereas it was not detected in PBLs prevented from making cell-to-cell contact. However, mRNA of HCMV IE was not detected in PBLs cultured with IL-2 and IFN-gamma that made direct contact with HCMV-infected MRC-5. Transmission of the pp65 antigen was increased in culture medium containing IL-4. At a higher viral infection titer, cell-free HCMV infected adherent PBLs cells. The subset, which did not adhere, did not infect cell-free viruses even at a very high multiplicity of infection. Moreover, the adhered subset of PBLs infected with HCMV was able to transmit HCMV to non-infected fibroblasts. Our results suggest that cell-to-cell contact (when PBLs make direct contact with HCMV-infected cells) is important in the mechanism of HCMV transmission and that the adherent cells of PBLs are one of the most important vehicles for HCMV infection. Moreover, we suggest that type 2 cytokines such as IL-4 enhance the transmission of HCMV to PBLs. PMID- 11402619 TI - Vascular endothelial growth factor (VEGF) serum concentration changes during chemotherapy in patients with lung cancer. AB - Vascular endothelial growth factor (VEGF) is one of the most important angiogenic factors. This study aimed at clarifying the clinical significance of the changes in the serum VEGF (S-VEGF) concentrations in patients with lung cancer during anticancer chemotherapy. Subjects comprised 29 patients with lung cancer (13 adenocarcinomas, 7 squamous cell carcinomas, and 9 small cell carcinomas) who were treated with cisplatin-based chemotherapy. S-VEGF was measured by ELISA. We compared the S-VEGF concentrations between the responders and nonresponders to anticancer chemotherapy. S-VEGF concentrations before treatment of the chemotherapy (pretreatment S-VEGF concentrations) were correlated with the number of WBC, neutrophil count, monocyte count and platelet count but not the lymphocyte count. The mean pretreatment S-VEGF concentrations in responders and those in nonresponders were not significantly different, 509.7 pg/ml in the former and 382.8 pg/ml in the latter, respectively., The S-VEGF concentrations in the responders decreased to a mean of 356.0 pg/ml and 304.1 pg/ml during and at the end of therapy, respectively while those in the nonresponders increased to a mean of 474.2 pg/ml and 598.4 pg/ml during and at the end of therapy. The S-VEGF concentration changes in the responders were significantly different from those in the nonresponders (p = .006). The S-VEGF concentration may relate to tumor burden, however it may not be a good marker for tumor burden, because it can be influenced by various factors such as neutrophil which increases during infection. A decrease in S-VEGF concentrations may improve neoangiogenesis and the immune response, and may correlate with improvements in the quality of life and survival of patients. PMID- 11402620 TI - Interleukin-4 induces mouse cytomegalovirus interstitial pneumonia in a latent infection model. AB - To better understand immune mechanisms involved in onset of cytomegalovirus pneumonia, we initially examined the replication of a low virulence strain of mouse cytomegalovirus (MCMV) in nude and BALB/c mice infected by intranasal inoculation. MCMV was detected by plaque assay in the salivary glands of nude mice from days 3 to 16, and in those of BALB/c mice from days 7 to 11. Nude mice became infected with MCMV earlier than BALB/c mice. Moreover, MCMV-DNA was detected in the salivary glands until day 16 after MCMV inoculation in nude and BALB/c mice. However, we did not find evidence of interstitial pneumonia at day 16 in either BALB/c or nude mice. These results suggest that this system represents a latent infection model in BALB/c mice and a persistent infection model in nude mice. We treated latently infected BALB/c mice with methylprednisolone or IL-4 every other day. The mice treated with IL-4 developed interstitial pneumonia, whereas those treated with m-PSL did not. In the present study, we constructed a model of MCMV latent infection that could be used to induce development of interstitial pneumonia. IL-4 appears to be a key cytokine for onset of interstitial pneumonia in mice with latent MCMV infection. PMID- 11402621 TI - Expression of the SART3 antigens in oral cancers. AB - It was previously reported that SART3 is a tumor-rejection antigen recognized by HLA-A24-restricted cytotoxic T lymphocytes (CTLs). This study investigated expression of the SART3 antigen in oral cancer as a candidate of tumor antigens for use in specific immunotherapy. The SART3 antigen was detected in all of the oral cancer cell lines tested, 24 of 31 (77%) oral cancer tissue samples, 0 of 3 oral benign tumors and 0 of 3 normal oral tissues. Oral cancer cell lines had the ability to stimulate IFN-gamma production by the HLA-A24-restricted and SART3 specific CTLs that were established from the peripheral blood mononuclear cells (PBMCs) of an esophageal cancer patient. Therefore, the SART3 antigen could be an appropriate vaccine candidate for a large number of oral cancer patients. PMID- 11402622 TI - Doppler echocardiographic studies of diastolic cardiac function in the human fetal heart. AB - Fetal echocardiography has been used for the noninvasive evaluation of human fetal cardiac anatomy, function, and hemodynamics. The purpose of the present study was to use Doppler echocardiographic methods to measure diastolic flow velocity patterns across the tricuspid and mitral valves in human fetuses during gestation. Fifty normal fetuses, 35 fetuses with intrauterine growth retardation (IUGR), and 30 fetuses of diabetic mothers (DM) were studied. Peak flow velocities during early diastole (peak E wave) and peak flow velocities during atrial contraction (peak A wave) were measured, and the peak E/A ratio was calculated. The peak E/A ratio of the left ventricle in fetuses increased gradually with increasing gestational age (r = 0.57, p < 0.05), and the peak E/A ratio of the right ventricle in fetuses increased linearly with increasing gestational age (r = 0.48, p < 0.05). In early gestational age, the peak E/A ratios of both the left and right ventricle in fetuses with IUGR and of DM were not significantly different from those in the controls. However, in late gestational age, the peak E/A ratios of both the left and right ventricle in fetuses with IUGR and of DM were significantly smaller than those in the controls. This gradual decrease in the peak E/A ratio during gestation may represent the maturational or developmental alteration of diastolic cardiac properties in utero. Fetuses with IUGR and of DM may have abnormal diastolic cardiac function in later gestation. PMID- 11402623 TI - Clinical significance of serum CD44 measurement in malignant lymphoma. AB - We monitored the effectiveness of lymphoma therapy by measuring the serum levels of soluble CD44std (sCD44std) and soluble CD44v6 (sCD44v6). Furthermore, we measured the level of soluble interleukin 2 receptor (sIL-2R). A total of 24 patients with non-Hodgkin's lymphoma were enrolled. sCD44std, sCD44v6, and sIL-2R on serum were measured using ELISA system. In all patients, only the sIL-2R level decreased significantly following therapy. However, an analysis of CR and PR showed that the degree of decrease in the sCD44std level was significantly greater than that in the sIL-2R level. Furthermore, among the CS IV cases, only the CD44std level decreased significantly after therapy. These findings suggest that the level of serum sCD44std reflects clinical pathology more closely than the level of serum sIL-2R in CS IV patients and those who respond well to therapy. Moreover, when T-cell and B-cell lymphomas were analyzed separately, the levels of sCD44std and sIL-2R decreased significantly after therapy in patients with B-cell lymphomas, and the degree of decrease in the sCD44std level was very significant with a p-value of 0.0003. This suggests that when sCD44std is used as an index of treatment, it more closely reflects the treatment of B-cell lymphomas. Level of serum sCD44std should prove to be a useful marker for assessing the effectiveness of lymphoma therapy. PMID- 11402624 TI - The biochemical study of intermaxillary fixation (IMF) stress in oral surgery inpatients. AB - Although intermaxillary fixation (IMF) is performed to treat the patients with maxillary fracture, this procedure is very stressful to the patients. IMF has been reported to increase noradrenaline (NA) release in the brain and elevate plasma corticosterone contents in the rat. These changes were significantly attenuated by diazepam, an anxiolytic of the benzodiazepine family. These results suggest that IMF could greatly affect the pituitary-adrenal system as a stress. In the present study, in order to examine the influence of IMF on the human body function, we measured levels of 17-hydrocorticosteroids (17-OHCS) and 17 ketosteroid (17-KS), which are metabolites of the adreno-cortical hormone cortisol, in the urine of inpatients undergoing IMF. The subjects were requested to fill out a questionnaire on irritableness caused by IMF. In these patients, urinary 17-OHCS levels were significantly increased after IMF and well correlated to the results of the questionnaire. The finding suggested that urinary 17-OHCS levels reflect stress related to IMF, and that such stress mainly causes an irritated feeling. Natural killer cell activity (NK activity), which is considered to be related to stress, was measured in these patients. The relationship between 17-OHCS levels and NK activity was examined in reference to the results of the questionnaire. Questionnaire showed that most patients noted insomnia and an irritated feeling during IMF. To examine the influence of anxiolytic agents on stress related to IMF, an anxiolytic agent, ethyl loflazepate, was administered during IMF, and urinary 17-OHCS levels were measured. There was no correlation between 17-OHCS levels and NK activity in the patients. Furthermore, no correlation was observed between visual analogue scale (VSA) and NK activity. Increases in 17-OHCS levels in the group treated with ethyl loflazepate, an anxiolytic of the benzodiazepine family, were significantly lower than in the untreated group. This suggests that ethyl loflazepate reduced stress responses to IMF. It has been reported that NK activity is reduced inpatients with depression or chronic fatigue syndrome. However, NK activity may not be affected by mechanical stress such as IMF. The finding that an anxiolytic agent, ethyl loflazepate, inhibited stress responses to IMF further suggests that anxiolytic drugs are very useful for treatment of irritated feeling of the patients undergoing IMF. PMID- 11402625 TI - Molecular genetic analysis of reovirus isolated in Korea. AB - Reovirus isolates from human, striped-field mouse (Apodemus agrarius) and Korean field mouse (A. peninsulae) in Korea showed extensive variability in the patterns of electrophoretic migration of the double-stranded RNA (dsRNA) genome segments. Hemagglutination inhibition (HI) test was performed for serotype determination of 12 reovirus isolates. To clarify genetic diversity and molecular phylogeny of Korean reoviruses, L1, S3 and S4 genomic segments of reoviruses were amplified by RT-PCR and directly sequenced. Among 12 reovirus strains, 9 strains were type 3 and 3 strains were type 2. The L1 was highly conserved showing 91.5-100%, 94.7 100% similarities among Korean isolates, and 77.5-97.9%, 92.6-96.8% similarities compared to other reference strains of each genotypes at nucleotide and amino acid levels, respectively. In S3 and S4 segments, 84.4-99.3%, 72.3-99% nucleotide sequence similarities and 92-99.3%, 89.1-98.4% amino acid sequence similarities among Korean isolates were observed, and 70.8-93.9%, 72.3-98.7% nucleotide sequence similarities and 81.8-100%, 88.3-97.7% amino acid sequence similarities compared with other reference strains of each genotype were observed, respectively. Phylogenetic analyses based on the S3 and S4 nucleotide sequences indicate that genotypes of reovirus are more related with geographic differences rather than host species or date of isolation. PMID- 11402626 TI - Fever and survival with drug-treated non-small cell lung cancer. AB - Patients with non-small cell lung cancer who develop fever during anti-neoplastic chemotherapy are treated empirically, before the cause of fever has been determined. Little study has been made of episodes of infection during chemotherapy with respect to antineoplastic response, other adverse reactions, or survival time. In particular, the relationship between fever and long-term survival is not clear. The prognostic significance of febrile episodes was investigated in patients with non-small cell lung cancer receiving full-dose chemotherapy. Most were treated only with chemotherapy during the period of investigation, although some had concurrent or subsequent radiotherapy, and a few had surgery following chemotherapy. No significant difference was seen between febrile and afebrile patients regarding therapeutic modality. Patients with poor performance status (PS) had a higher incidence of febrile episodes (P < .001). Multivariate analysis, however, showed febrile episodes to be a major prognostic factor independent of PS on clinical stage. PS and stage affected short-term survival, while febrile episodes were related to decreased long-term survival (203 vs. 366 days; with deaths in the first 3 months excluded, 241 vs. 377 days). Although existing evidence is inconclusive, our study suggests that prevention of infection during antineoplastic treatment by administration of granulocyte stimulating factors and early initiation of antimicrobials might improve long term survival. PMID- 11402627 TI - Role of veterinary medicine in public health: antibiotic use in food animals and humans and the effect on evolution of antibacterial resistance. AB - Veterinary public health is another frontier in the fight against human disease. The veterinary public health scope includes the control and eradication of zoonoses, diseases that are naturally transmitted between vertebrate animals and man. These diseases pose a continuous hazard to the health and welfare of the public. More than 100 diseases are categorized as zoonoses, including salmonellosis. It is important to understand how antibiotics are used in humans and in food animals and how these uses affect the evolution of antibacterial resistance. Appropriate use of antibiotics for food animals will preserve the long-term efficacy of existing antibiotics, support animal health and welfare, and limit the risk of transfer of antibiotic resistance to humans. An understanding of the epidemiology of antimicrobial resistance allows development of preventive strategies to limit existing resistance and to avoid emergence of new strains of resistant bacteria. Risk assessments are being used by the Center for Veterinary Medicine at the U.S. Food and Drug Administration as regulatory tools to assess potential risk to humans resulting from antibiotic use in food producing animals and to then develop microbial safety policies to protect the public health. The veterinary public health scope, in addition to the control and eradication of zoonoses, also includes the development and supervision of food hygiene practices, laboratory and research activities, and education of the public. Thus, it may be seen that there are many ways in which veterinary medicine plays a very important role in public health. PMID- 11402628 TI - The effect of 3-month ingestion of Ginkgo biloba extract (EGb 761) on pancreatic beta-cell function in response to glucose loading in individuals with non-insulin dependent diabetes mellitus. AB - In the first report (Journal of Clinical Pharmacology 2000; 40:647-654), it was shown that ingestion of 120 mg of Ginkgo biloba extract (EGb 761) daily for 3 months by normal glucose-tolerant individuals caused a significant increase in pancreatic beta-cell insulin and C-peptide response, measured as the area under the curve (AUC0-->120) during a 2-hour standard (75 g) oral glucose tolerance test (OGTT). This follow-up study was designed to determine the effect of the same Ginkgo biloba treatment on glucose-stimulated pancreatic beta-cell function in non-insulin-dependent diabetes mellitus (NIDDM) subjects. In diet-controlled subjects (fasting plasma glucose [FPG], 117 +/- 16 mg/dl; fasting plasma insulin [FPI], 29 +/- 8 microU/ml; n = 6), ingestion of Ginkgo biloba produced no significant effect on the insulin AUC0-->120 (193 +/- 53 vs. 182 +/- 58 microU/ml/h, before and after ingesting Ginkgo biloba, respectively). In hyperinsulinemic NIDDM subjects taking oral hypoglycemic medications (n = 6) (FPG 143 +/- 48 mg/dl; FPI 46 +/- 13 microU/ml), ingestion of Ginkgo biloba caused blunted plasma insulin levels from 30 to 120 minutes during the OGTT, leading to a reduction of the insulin AUC0-->120 (199 +/- 33 vs. 147 +/- 58 microU/ml/h, before and after Ginkgo biloba, respectively). The C-peptide levels increased, and so the AUC0-->120 did not parallel the insulin AUC0-->120, creating a dissimilar insulin/C-peptide ratio indicative of an enhanced hepatic extraction of insulin relative to C-peptide. Thus, in pancreatic beta-cells that are already maximally stimulated, ingestion of Ginkgo biloba may cause a reduction in plasma insulin levels. Only in NIDDM subjects with pancreatic exhaustion (FPG 152 +/- 46 mg/dl; FPI 16 +/- 8 microU/ml; n = 8), who also took oral hypoglycemic agents, did Ginkgo biloba ingestion significantly increase pancreatic beta-cell function in response to glucose loading (insulin AUC0-->120 increased from 51 +/- 29 to 98 +/- 20 microU/ml/h, p < 0.0001), paralleled by a C-peptide AUC0-->120 increase from 7.2 +/- 2.8 to 13.7 +/- 6.8 (p < 0.0001). Whether this increase is due to "resuscitation" of previously exhausted islets or increased activity of only the remaining functional islets is unclear. However, not even in this group did increased pancreatic beta-cell activity cause a reduction of blood glucose during the OGTT. It is concluded that ingestion of Ginkgo biloba extract by an NIDDM subject may increase the hepatic metabolic clearance rate of not only insulin but also the hypoglycemic agents. The result is reduced insulin-mediated glucose metabolism and elevated blood glucose. PMID- 11402629 TI - Montelukast dose selection in children ages 2 to 5 years: comparison of population pharmacokinetics between children and adults. AB - Montelukast, a leukotriene receptor antagonist, has demonstrated efficacy and tolerability in the treatment of asthma in patients age 6 years and older. The purpose of this open, one-period, multicenter population pharmacokinetic study was to identify a chewable tablet (CT) dose of montelukast for administration to children ages 2 to 5 years with asthma, yielding a single-dose pharmacokinetic profile (area under the plasma concentration-time curve [AUC]) comparable to that of the 10 mg film-coated tablet (FCT) dose in adults. Because patient numbers were small and the volume of blood that could be collected from individual 2- to 5-year-old patients was limited, a population pharmacokinetic approach was used to estimate population AUC (AUCpop). The 4 mg CT dose of montelukast was well tolerated and yielded an AUCpop (2721 ng.h/mL) similar to that of the adult AUCpop (2595 ng.h/mL) observed after a 10 mg FCT dose. These results support the selection of a 4 mg once-daily CT dose of montelukast for future efficacy and safety studies in children ages 2 to 5 years with asthma. PMID- 11402630 TI - Caffeine metabolism in premature infants. AB - Caffeine has been used frequently in the treatment and prevention of apnea of prematurity. The metabolism of caffeine depends on the activities of the hepatic enzymes that vary from one infant to another. The objective of this study was to determine the influence of postnatal age (PNA), birth weight (BW), study weight (SW), gestational age (GA), postconceptual age (PCA), and gender on the maturation of caffeine metabolism in premature infants. The caffeine base was administered orally as a loading dose of 10 mg/kg, followed by a maintenance dose of 2 mg/kg every 24 hours. The steady-state concentration of caffeine and metabolites was measured in plasma taken on the 5th-day postloading dose. The molar concentration ratios for the N3 (N3-), N7 (N7-), N1 (N1-), and all methyl (Nall-) demethylation processes; clearance (CL); and the percentage of molar concentration of caffeine found in plasma to that of the total caffeine and metabolites (%CAF) were calculated from samples collected from 80 neonatal infants. The 48 male and 32 female premature infants had median (range) BW (g), GA (weeks), SW (g), PCA (weeks), and PNA (days) of 1300 (650-2260), 30 (24-34), 1630 (980-2670), 34 (29-40), and 28 (5-60), respectively. The median (range) of the ratios for the %CAF, CL, and the N3-, N7-, N1-, and Nall- were 86.9 (52.9 99.0), 0.127 (0.046-0.503) ml.kg-1.min-1, 0.032 (0-0.438), 0.070 (0.007-0.471), 0.026 (0-0.283), and 0.0463 (0.003-0.303), respectively. When the patients were stratified into four PNA age groups, each older group showed a consistently higher level of caffeine metabolic activity for the N3-, N7-, and Nall- pathways with a corresponding decrease in the %CAF, whereas no significant differences were seen for the N1-pathway or for CL. No pattern of significant differences between the demethylation process ratios, %CAF, or CL was seen between groups of infants when they were stratified according to BW, SW, PCA, or GA. The female infants were found to have significantly higher rates of caffeine metabolism as shown by %CAF, N1-, N3-, and Nall- processes but not the N7-. Multivariate linear regression analysis by two methods demonstrated that PNA is significantly related to %CAF and Nall-, whereas the female patients had higher levels of metabolic activity for the %CAF and N1- process. The authors conclude that the N7-demethy lation process is the predominate caffeine metabolic process in premature infants. Furthermore, the maturation of the caffeine metabolism in premature infants with a PNA of less than 60 days increases with postnatal age, regardless of birth weight, gestational age, postconceptual age, and study weight. The female neonatal patients demonstrated a higher rate of caffeine metabolism than the males. PMID- 11402631 TI - Comparative tacrolimus pharmacokinetics: normal versus mildly hepatically impaired subjects. AB - Tacrolimus (FK506, Prograf), marketed for the prophylaxis of organ rejection following allogenic liver or kidney transplantation, is virtually completely metabolized. The major metabolic pathways are P450 3A4-mediated hydroxylation and demethylation. Since P450 hepatic drug-metabolizing enzymes may be impaired in hepatic dysfunction, a study was conducted to characterize oral and intravenous tacrolimus pharmacokinetics in 6 patients with mild hepatic dysfunction and compared with parameters to those from normal subjects obtained in a separate study. Patients received two treatments: a single 0.020 mg/kg ideal body weight (IBW) i.v. dose infused over 4 hours and approximately 0.12 mg/kg IBW orally; normal subjects were dosed at 0.02 mg/kg 4-hour i.v. and 5 mg (0.065 mg/kg) p.o. Mean blood pharmacokinetic parameters with mild hepatic dysfunction were as follows: clearance = 0.035 L/h/kg, terminal exponential volume of distribution = 2.59 L/kg, terminal exponential half-life = 60.6 hours (i.v.), p.o. maximum blood concentration = 48.2 ng/mL, time of p.o. maximum blood concentration = 1.5 hours, and absolute bioavailability = 22.3%. The respective parameters in normal subjects were as follows: 0.040 L/h/kg, 1.91 L/kg, 34.2 hours (i.v.), 29.7 ng/mL, 1.6 hours, and 17.8%. Inasmuch as clearance and bioavailability were not substantially different from that in normal subjects, patients with mild hepatic impairment may initially be treated with conventional tacrolimus doses, with subsequent dosage adjustments based on response, toxicity, and therapeutic drug monitoring. PMID- 11402632 TI - Effects of tolterodine, trospium chloride, and oxybutynin on the central nervous system. AB - Antimuscarinic compounds are increasingly used to treat the symptoms of overactive bladder; however, their use is often restricted by peripheral adverse effects (AEs). On the other hand, data regarding their influence on the central nervous system (CNS) are limited. This randomized, single-blind, parallel-group quantitative-topographical EEG (qEEG) study of clinical phase I investigates the potential CNS adverse effects of the three antimuscarinic drugs--tolterodine, oxybutynin, and trospium chloride--in comparison to placebo. Overall, 4 x 16 (total 64) young, healthy male volunteers were included in the study. The subjects were given either placebo or the clinically recommended daily doses of the drugs dispensed in three doses on a single day (tolterodine 2 mg bid and once placebo, total 4 mg/d; oxybutynin 5 mg tid, total 15 mg/d; and trospium chloride 15 mg tid, total 45 mg/d). The qEEG was recorded prior to and up to 4 hours after each intake of the trial medication (a total of 10 qEEG sessions) under three different conditions: at rest with eyes open, eyes closed, and under mental demand. The drug tolerability was subjectively evaluated by the volunteer and the investigator. In comparison to placebo (10% confidence interval), tolterodine and trospium chloride did not induce changes of the qEEG power in five of the six frequency bands (i.e., delta, alpha 1, alpha 2, beta 1, and beta 2). Isolated power decreases were only observed in the theta frequency band. In contrast, oxybutynin caused significant power reductions in four frequency bands (theta, alpha 1, alpha 2, and beta 1; p < 0.01). The subjectively evaluated drug tolerability was comparable between all treatment groups, although differences in the AE occurrence existed, with the AE frequency being higher in the oxybutynin group. The results of this study support the findings that oxybutynin as a tertiary amine crosses the blood-brain barrier, causing significant qEEG activity changes and more pronounced central adverse effects. Although tolterodine is also a tertiary amine, it shows limited effects on qEEG activity (i.e., slight theta power reductions), comparable to the effects of trospium chloride, a quarternary amine, which barely crosses the blood-brain barrier. The minimal qEEG changes observed with tolterodine and trospium chloride reflect most probably a rebound message from the peripheral target organs. Prescription of oxybutynin thus implicates a higher risk of CNS side effects. PMID- 11402633 TI - The pharmacokinetics of colistin in patients with cystic fibrosis. AB - The safety and pharmacokinetics of colistin were determined after first dose (n = 30) and again under steady-state conditions (n = 27) in 31 patients with cystic fibrosis receiving the drug as a component of their treatment for an acute pulmonary exacerbation of their disease. Patients ranged in age from 14 to 53 years and received colistin for 6 to 35 days. Each patient was started on colistin 5 to 7 mg/kg/day administered intravenously in three equally divided doses. Elimination half-life (t1/2), mean residence time (MRT), steady-state volume of distribution (Vdss), total body clearance (Cl), and renal clearance (Clr) after first-dose administration averaged 3.4 hours, 4.4 hours, 0.09 l/kg, and 0.35 and 0.24 ml/min/kg, respectively. No differences in colistin disposition characteristics between first-dose and steady-state evaluations were observed. Sputum sampling was incomplete and confounded by previous aerosol administration but revealed colistin concentrations that markedly exceeded observed plasma concentrations. Twenty-one patients experienced one or more side effects attributed to colistin administration. The most common reactions involved reversible neurologic manifestations, including oral and perioral paresthesias (n = 16), headache (n = 5), and lower limb weakness (n = 5). All of these apparent colistin-induced neurologic adverse effects, though bothersome, were benign and reversible. Intermittent proteinuria was observed on urinalysis in 14 patients, and 1 patient developed reversible, colistin-induced nephrotoxicity. No relationship between the occurrence of any colistin-associated adverse effect and plasma colistin concentration or colistin pharmacokinetic parameter estimate was observed. These data provide no basis for routine monitoring of colistin plasma concentrations to guide dosing for patient safety and suggest slow upward dose titration to minimize the incidence and severity of associated side effects. PMID- 11402634 TI - Pharmacokinetics of metformin gastric-retentive tablets in healthy volunteers. AB - The single-dose pharmacokinetics of two gastric-retentive, extended-release tablet formulations of metformin hydrochloride in fed, healthy volunteers were compared with those of the currently marketed immediate-release metformin hydrochloride product. The plasma concentration-time profiles demonstrated extended-release characteristics from the gastric-retentive tablets. The mean bioavailability from each gastric-retentive tablet was approximately 115%, relative to the immediate-release (IR) product. Cmax values were lower and tmax values were greater for the gastric-retentive tablets compared with the IR product. In contrast to conventional extended-release metformin tablets reported in the literature, these gastric-retentive tablets showed extended-release plasma concentration profiles of metformin hydrochloride and increased bioavailability compared with the immediate-release tablet. PMID- 11402635 TI - Thalidomide dose proportionality assessment following single doses to healthy subjects. AB - Thalidomide is approved in the United States for treating erythema nodosum leprosum, a complication of leprosy. The present study determined the single-dose oral pharmacokinetics and dose proportionality from 50 to 400 mg of Celgene's commercial Thalomid thalidomide formulation in an open-label, single-dose, three way crossover study. Fifteen healthy subjects were given 50, 200, and 400 mg of thalidomide on three occasions, and blood samples were collected over 48 hours. Pharmacokinetic parameters were determined using noncompartmental methods, and dose proportionality was assessed by linear regression of dose-normalized Cmax and AUC0-infinity. No serious or unexpected adverse events occurred. The most common adverse events were dizziness, somnolence, headache, and nausea. One patient was discontinued because of pharyngitis. There was a significant deviation from proportionality for Cmax with increases being less than proportional than changes in dose. AUC0-infinity increased proportionally with dose, suggesting that the overall amount of thalidomide absorbed, as well as its clearance, is independent of dose over the range used. V/F was found to increase with dose. This was most likely due to the terminal rate constant, which is used to calculate V/F, actually representing the absorption process rather than elimination (i.e., flip-flop phenomenon). The terminal rate constant (absorption rate constant) for the highest dose was 50% less than for the other two lower doses. The less than proportional increases in Cmax were most likely due to thalidomide's low aqueous solubility. Thalidomide shows reasonable dose proportionality with respect to AUC from 50 to 400 mg. PMID- 11402636 TI - Pharmaceutical care services for asthma patients: a controlled intervention study. AB - As asthma is associated with an enormous social, psychological, and economic burden, various patient education programs have been developed to improve outcomes, including quality of life. The authors evaluated the effectiveness of community pharmacy-based interventions on lung function, health-related quality of life, and self-management in asthma patients in a 12-month controlled intervention study in 26 intervention and 22 control pharmacies. Pharmacies opted whether to take part as intervention or control pharmacies. According to this, patients (ages 18-65) with mild to severe asthma attending the pharmacies were allocated to the intervention (n = 161) or control group (n = 81), respectively. Intervention patients were educated on their disease, pharmacotherapy, and self management; inhalation technique was assessed and, if necessary, corrected. Pharmaceutical care led to significantly improved inhalation technique. Asthma specific quality of life and the mental health summary score of the SF-36 improved significantly in the intervention group. At 12 months, the intervention group showed significant improvements with regard to evening peak flow, self efficacy, and knowledge. PMID- 11402637 TI - A pilot study assessing the impact of heat on the transdermal delivery of testosterone. AB - This study evaluated the effect of locally applied heat on the transdermal delivery of testosterone. Six healthy adult volunteers were tested three times while receiving a 5 mg androgen patch, the same patch with a heat-generating patch, and no patch at all over 12 hours. Statistically significant differences in mean maximum serum testosterone concentration values were seen. Heat plus patch resulted in a mean maximum serum testosterone concentration of 939 ng/dl versus 635 ng/dl (patch only) and 425 ng/dl (no patch). (Heat + patch vs. no patch: p < 0.001; heat + patch vs. patch: p < 0.001; patch vs. no patch: p = 0.003.) The area under the curve of plasma testosterone concentration versus time values were means of 4114 ng/dl.h versus 1985 ng/dl.h for the patch-only group (p = 0.001). The use of heat improved absorption of transdermal testosterone and decreased time to peak serum testosterone concentrations, resulting in a statistically significant difference in mean maximum serum testosterone concentrations compared with the use of the patch without heat. PMID- 11402638 TI - Lack of effect of rosiglitazone on the pharmacokinetics of oral contraceptives in healthy female volunteers. AB - The effect of rosiglitazone (Avandia [BRL 49653C]) on the pharmacokinetics of ethinylestradiol and norethindrone was evaluated after repeat dosing of rosiglitazone with an oral contraceptive (OC; Ortho-Novum 1/35 containing norethindrone 1 mg and ethinylestradiol 0.035 mg) in a randomized, double-blind, placebo-controlled crossover study. Thirty-four healthy female volunteers received oral rosiglitazone (RSG) 8 mg + OC or matched placebo (P) + OC daily on days 1 to 14 of a 28-day OC dosing cycle; the alternate regimen was administered during a second cycle. Ethinylestradiol and norethindrone pharmacokinetics were determined from plasma concentrations on day 14. Lack of pharmacokinetic effect was prospectively defined as 90% CI for the point estimate (PE) of the ratio (RSG + OC):(P + OC) contained within a 20% equivalence range for both ethinylestradiol and norethindrone (analyzed by ANOVA). For RSG + OC and P + OC, respectively, mean ethinylestradiol AUC(0-24) was 1126 and 1208 pg.h/mL (PE: 0.92; 90% CI: 0.88 0.97), and mean norethindrone AUC(0-24) was 178 and 171 ng.h/mL (PE: 1.04; 90% CI: 1.00-1.07). Thus, rosiglitazone had no significant effects on the pharmacokinetics of ethinylestradiol or norethindrone. Coadministration of rosiglitazone with OCs does not induce metabolism of these synthetic sex steroids and is not expected to impair the efficacy of OCs or hormone replacement therapy. PMID- 11402639 TI - Low molecular weight heparin-induced liver toxicity. AB - The authors report two cases of hepatotoxicity induced by low molecular weight heparin. A 26-year-old woman and a 33-year-old man were treated with low molecular weight heparin for pulmonary embolism and cerebral infarction, respectively. They both developed derangement in liver function tests a few days after commencement of the low molecular weight heparin. The derangement in liver function tests was associated with a decreased serum complement 3 activity. Their liver functions recovered over a period of 2 to 3 months after low molecular weight heparin was stopped. Liver biopsy in the woman demonstrated balloon degeneration with scattered foci of hepatocytic necrosis, suggesting a complement mediated hepatocellular damage. PMID- 11402640 TI - Characterisation of some Ornithobacterium rhinotracheale strains and examination of their transmission via eggs. AB - The biochemical characteristics and antibiotic susceptibility of 12 Ornithobacterium rhinotracheale strains isolated from chickens and turkeys suffering from respiratory clinical signs and the survival of some isolates on egg-shell and within chicken eggs during hatching were examined. All O. rhinotracheale strains showed typical biochemical characteristics. Among the 16 drugs examined, penicillin G, ampicillin (MICs ranging from < or = 0.06 microgram/ml to 1 microgram/ml), ceftazidim (with MICs from < or = 0.06 microgram/ml to 0.12 microgram/ml), erythromycin, tylosin, tilmicosin (with some exceptions MICs ranged from < or = 0.06 microgram/ml to 1 microgram/ml) and tiamulin (MICs varied from < or = 0.06 microgram/ml to 2 micrograms/ml) were the most effective. Lincomycin, oxytetracycline and enrofloxacin also gave good inhibitions, but with most strains in a higher concentration (MICs ranged in most cases from 2 micrograms/ml to 8 micrograms/ml). The other antibiotics inhibited the growth of O. rhinotracheale only in very high concentrations (colistin) or not at all (apramycin, spectinomycin, polymyxin B). At 37 degrees C, O. rhinotracheale did not survive on egg-shell for more than 24 hours, while upon inoculation into embryonated chicken eggs it killed embryos by the ninth day, and from the 14th day post-inoculation no O. rhinotracheale could be cultured from the eggs at all. These results suggest that O. rhinotracheale is not transmitted via eggs during hatching. PMID- 11402641 TI - Comparison of erythromycin and oxytetracycline for the treatment of ovine footrot. AB - A microbiological study of 25 cases of ovine footrot was performed. Cultures belonging to Dichelobacter nodosus were isolated in 48% of the sampled animals. The sensitivity of the 99 strict anaerobic bacterial isolates to 5 antibiotics (penicillin G, amoxycillin, spiramycin, erythromycin and oxytetracycline) was studied. The percentage of resistant cultures was in all cases higher than 30%. The efficacy of erythromycin and oxytetracycline in the treatment of ovine footrot was studied. To conduct this test, an intramuscular injection was applied, of one antimicrobial or the other, at the beginning of the treatment. The tolerance of animals to the antimicrobials, the success rate of treatment and the severity of lameness were evaluated. The percentage of animals cured within 15 days was around 75%. In contrast, only 44% improvement was achieved in the lameness. No differences were found between the two antimicrobials in the above indices. PMID- 11402642 TI - Ultrasonography and surgery of canine biliary diseases. AB - Findings of hepatic and gallbladder ultrasonography were analyzed in 12 dogs with gallbladder and/or extrahepatic biliary tract obstruction and compared with the results of exploratory laparotomy. Hepatic ultrasonography demonstrated normal liver in 2 dogs and hepatic abnormalities in 10 animals. The following ultrasonographic diagnoses were established compared to surgical findings: gallbladder obstruction caused by bile sludge (correct/incorrect: 1/2, surgical diagnosis: choleliths in one case), gallbladder obstruction caused by neoplasm (0/1, surgical diagnosis: mucocele), gallbladder and extrahepatic biliary tract obstruction due to choleliths (3/3), extrahepatic biliary tract obstruction caused by pancreatic mass (1/1) and small intestinal volvulus (1/1). Bile peritonitis caused by gallbladder rupture (4/4) was correctly diagnosed by ultrasound, aided with ultrasonographically-guided abdominocentesis and peritoneal fluid analysis. Rupture of the gallbladder should be suspected in the presence of a small, echogenic gallbladder or in the absence of the organ together with free abdominal fluid during ultrasonography. Laparotomy was correctly indicated by ultrasonography in all cases. However, the direct cause of obstruction could not be determined in 2 of the 12 dogs by ultrasonography alone. PMID- 11402643 TI - Bacteriological recovery after intramuscular or intracisternal spiramycin-based drying-off therapy. AB - Systemic (intramuscular, i.m.) vs. local (intracisternal, IC) routes of spiramycin-based drying-off therapy were compared for efficacy on 65 Staphylococcus aureus infected udder quarters of 38 dairy cows. Single-dose (30,000 IU/kg) i.m. treatment (single i.m. group) resulted in a similarly low bacteriological recovery rate (14%) as seen in the untreated controls (18%). I.m. treatment (30,000 IU/kg) on 4 consecutive days (4 i.m. group) resulted in significantly higher quarter-based recovery rates than that in the single i.m. group. The bacteriological recovery rates obtained in the intracisternal and 4 i.m. groups were quite similar but remained below 50%. Based on these findings as well as on the high cost of the repeated intramuscular treatment regime there is no reason to give extra preference to the systemic application of spiramycin at drying off in the practice. PMID- 11402644 TI - Histological evaluation of immune organs in chicken embryos inoculated with Marek's disease virus and lymphokines. AB - The aim of the present study was to evaluate the presence of lymphocytes and granulocytes in different stages of embryonic development and on the first posthatching day. The lymphocytes present in the bursa of Fabricius and thymus were evaluated by histological analysis of the yolk sac, bursa of Fabricius, thymus, liver and bone marrow of 100 chicken embryos divided into groups and treated with: (I) Marek's disease vaccine as viral antigen, (II) Marek's disease vaccine plus lymphokines, (III) lymphokines, and (IV) vaccine diluent. Group V was not treated. Samples were taken on days 14, 17 and 20 of incubation and on the first posthatching day. An increase in the number of epithelial matrix as precursors of lymphoid follicles was observed in the bursa of Fabricius of embryos inoculated with lymphokines compared to embryos in all the other groups (p < 0.05). In addition, a higher amount of granulocytes was found in the yolk sac and liver of embryos inoculated with lymphokines than in the embryos of all other groups (p < 0.05). In the bone marrow, no significant difference was observed among the treated groups concerning the amount of granulocytes. The results suggest that administration of antigens or protein molecules at an early stage of embryonic development increases the presence of granulocytes in the liver and granulopoiesis in the yolk sac, and also increases the number of epithelial matrixs in the bursa of Fabricius. PMID- 11402645 TI - Characterisation and effects of a xylanase enzyme preparation extracted from Thermomyces lanuginosus cultures. AB - This paper describes the production of an enzyme preparation from the fungus Thermomyces lanuginosus. Thermal resistance, pH stability and lignocellulolytic activity of the enzyme preparation high in xylanase were studied on a variety of grains and forages. The enzyme preparation preserved more than 70% of its original xylanase activity for 4 and 1 h at 60 and 70 degrees C, respectively. The xylanase activity remained over 80% when the preparation was incubated for 30 min at pH 4.5. In vitro digestibility studies indicated that the enzyme digested 7.5, 8.5 and 8.0% of the dry matter (DM) of barley meal, wheat bran and oat meal samples, respectively. When applying 60-min incubation, 7.5, 7.3 and 8.4% of DM of the oat straw, alfalfa hay and triticale straw was digested, respectively. When the time of digestion was increased to 360 min, the sunflower hull showed 15.8% DM digestibility. PMID- 11402646 TI - Bovine abortion associated with Neospora caninum in Hungary. AB - The authors report the first case of bovine neosporosis in Hungary based on investigations made on an aborted fetus. The diagnostic methods included traditional as well as molecular techniques. This record extends further the geographic range of the disease. PMID- 11402647 TI - Identification of Trypanosoma evansi by DNA hybridisation using a non-radioactive probe generated by arbitrary primer PCR: short communication. AB - A highly reproducible, dominant, monomorphic fragment of 473 base pair (bp) amplified from the genome of Trypanosoma evansi by arbitrary primer-polymerase chain reaction (AP-PCR) was labelled with digoxigenin and investigated for its potential as DNA probe. Dot-blot hybridisation of total genomic DNA with the probe proved useful in detecting bubaline, cameline and equine strains of T. evansi down to 10 pg of parasite template DNA. No cross-hybridisation was seen with Babesia bigemina, Theileria annulata and the bubaline host DNA. This probe may facilitate laboratory identification of T. evansi in developing countries, without the inherent risk associated with radioisotopes. PMID- 11402648 TI - Rate of lipid peroxidation in brain and liver tissues and the total antioxidant status of blood plasma in developing chicks. AB - Age-related changes of tissue lipid peroxidation (LPO) of liver and brain, as well as plasma antioxidant capacity of broiler chicken cockerels were investigated. Tissue LPO was characterised by the spectrophotometric assessment of thiobarbituric acid reactive substances (TBARS). Plasma antioxidant power was evaluated by the measurement of total antioxidant status (TAS). Newly hatched broiler chicks had similar TAS value (1.19 mmol/l) as newborns of mammalian species. Significant changes (p < 0.05) were observed in the time course of all parameters. Tissue TBARS concentration was higher in the brain than in the liver at hatching, while the latter organ was found to have more effective antioxidant defence during embryonic life. The concentration of TBARS increased up to the 10th day in the liver but only up to the 21st day in the brain, and the former was accompanied by an approximately 50% decrease of plasma antioxidant capacity. This suggests that the liver plays an important role in forming the antioxidant defence mechanisms of the blood plasma in broiler chicks. PMID- 11402649 TI - Absorption of inorganic, trivalent and hexavalent chromium following oral and intrajejunal doses in rats. AB - The intestinal absorption of trivalent and hexavalent chromium (Cr) given orally (experiment I) or infused in the intestine (experiment II) was investigated in rats. The nonabsorbable form of chromium (51Cr2O3) and water-soluble and more absorbable Na2(51)CrO4 (the hexavalent form of Cr) were compared. Total retention of chromium given orally ranged around 15 percent of the dose, regardless of the chromium compounds applied. The absorption rate of chromic oxide, which is considered a nonabsorbable compound, was 14.4 as a percentage of chromium intake. This result indicates that some loss of chromium has to be taken into account in metabolic trials made by the indicator method. In isolated rat intestine, from the injected Cr 2.5% of chromic oxide and 43.2% of sodium chromate were absorbed during an hour (experiment II). The absorbed chromium was transferred to the liver where the liver tissue retained 10.9% of chromic oxide and 51.1% of sodium chromate. Radioactivity of v. cava caudalis following intestinal injection of Na2CrO4 was thirtyfold greater than after Cr2O3 dosing. This phenomenon can be explained by the lower blood clearance of chromate. Different absorption rate of chromate depending on the route of administration could be due to the fact that the hexavalent form given orally was reduced to Cr3+ in the acidic environment of the stomach. When Na2CrO4 was infused directly in the intestine of rats, such reduction could not occur. This means that the acidic gastric juice might play a role in inhibiting the intestinal absorption of Na2CrO4 when this compound is given orally. PMID- 11402650 TI - Effect of low-dose zearalenone exposure on luteal function, follicular activity and uterine oedema in cycling mares. AB - The effect of 10-day zearalenone administration starting 10 days after ovulation was studied in 6 cycling trotter mares in the summer period. After an entire oestrous cycle (Cycle 1), mares were given 7 mg purified zearalenone per os daily (1 mg/ml in ethyl alcohol) beginning on Day 10 of Cycle 2. Toxin exposure was continued until the subsequent ovulation. Luteal function and follicular activity were monitored daily by rectal palpation, ultrasonography and blood sampling for progesterone. During toxin exposure, all animals were in good physical condition. The toxin had no effect on the length of the interovulatory intervals, luteal and follicular phases. It did not influence significantly the plasma progesterone profiles (logistic curve parameters A1 to A6), the follicular activity (growth rate, maximum size of the ovulatory follicles, maximum number and the time of first increase in the number of large follicles) and the uterine oedema. It is concluded that in cyclic mares the methods used in this study could not detect any adverse effect of zearalenone (administered at a low dose similar to natural exposure) on reproduction. PMID- 11402651 TI - Evaluation of membrane integrity of frozen/thawed deer spermatozoa: short communication. AB - A simultaneous live/dead and acrosome staining, originally described for domestic mammals, was successfully applied on red deer (Cervus elaphus) and fallow deer (Dama dama) spermatozoa collected from the cauda epididymidis and vas deferens of shot stags. The staining is simple enough for routine application. Seven classes of spermatozoa were distinguished in the smears of frozen/thawed semen samples. Morphology, including cytoplasmic droplets, was evaluated as well. Percentage of live cells with intact acrosomes and with no other morphological aberrations might be a practical index of semen quality. PMID- 11402652 TI - Therapeutic efficacy of oxime reactivators in fenitrothion toxicity in buffalo calves (Bubalus bubalis). AB - The therapeutic efficacy of 2-pyridine aldoxime methochloride and diacetylmonoxime (DAM) alone and in combination with atropine was determined in oral fenitrothion toxicity in buffalo calves. DAM alone and in combination with atropine constitute the most effective therapy of fenitrothion poisoning. As compared to 2-pyridine aldoxime methochloride, DAM was also more effective in reactivating the fenitrothion-inhibited erythrocyte and plasma acetylcholinesterase and serum carboxylesterase enzymes and reversing fenitrothion-induced hyperglycaemia, hyperproteinaemia and hypercreatinaemia in animals. PMID- 11402653 TI - Sequence analysis of the membrane protein gene and nucleocapsid gene of porcine reproductive and respiratory syndrome virus isolated from a swine herd in Hungary. AB - Porcine reproductive and respiratory syndrome virus (PRRSV) was isolated from blood samples taken at a pig farm in Hungary from pigs showing clinical signs of the disease. The virus (ABV 32) was identified as belonging to the European genotype by using type-specific monoclonal antibodies. This was confirmed by comparing the sequence of the membrane protein gene (ORF 6) and the nucleocapsid gene (ORF 7) with the American VR2332 and the European LV genotype reference strain, respectively. Analysis of the amino acid sequence of the ORF 6 and ORF 7 of ABV 32 revealed five amino acid changes in both ORFs when compared with LV, of which two changes in ORF 7 were only found in the Spanish isolates. Additionally, the ORF 7 sequence was compared with corresponding sequences of a total of 21 other European strains. Phylogenetic analysis using the PHYLIP package confirmed the close relationship between the Hungarian and the Spanish isolates. Of all the isolates analysed, ABV 32 and LV were the least related. PMID- 11402654 TI - Characterisation of the fiber gene and partial sequence of the early region 4 of bovine adenovirus 2 (short communication). AB - The full sequence of the fiber gene and partial sequence of the putative 17 kD protein gene of bovine adenovirus-2 (BAdV-2) were determined. The size of the fiber gene of BAdV-2 proved to be 561 amino acids, of which the amino acids 37 to 385 form a typical shaft domain of 22 repetitive motifs. On the complementary strand, a gene homologous to the 17 kD protein coded in the E4 region of several human adenoviruses was found. The sequence analysis seems to confirm the presence of an intron in the sequenced part of the E4 region. PMID- 11402655 TI - Screening of Hungarian cattle herds for Mycoplasma mycoides subspecies mycoides small colony infection with negative results. AB - At abattoirs and farms, 1248 sera were collected from animals representing 121 farms, and examined by complement fixation test using Mycoplasma mycoides subspecies mycoides small colony type (MmmSC) antigen. All sera were negative except seven from four farms, giving ++ reactions in the serum dilution of 1:10. On retesting, these sera and additional 30 sera collected repeatedly in both farms gave negative results. In isolation attempts, 953 lung samples collected from slaughtered cattle at the same abattoirs, and 326 nasal swabs collected from 11 herds proved to be negative for the presence of MmmSC, but M. bovis was isolated frequently. In the small farms 23.95% of the animals had pleurisy and/or pneumonia while in the large herds 34.69% had lesions. DNA extracted from 50 nasal swabs and 430 lung samples was examined by polymerase chain reaction (PCR) using M. mycoides cluster-specific primers. DNA from further 325 lung samples was tested by the more specific M. mycoides subspecies mycoides small colony/large colony/capri specific primers and 196 samples by nested PCR specific for MmmSC. All gave negative results. The detection level of cluster-specific primers and the more specific primers was 33.4 pg of DNA, whereas that of nested PCR was 0.33 pg. PMID- 11402656 TI - Clinical study of the disease of calves associated with Mycoplasma bovis infection. AB - Clinical, bacteriological and serological examination of 35 calves from the age of 5 to 26 days was performed in a Holstein-Friesian dairy herd endemically infected with Mycoplasma bovis. M. bovis was isolated from 48.6% of nasal swabs taken from the calves at the age of 5 days, and from 91.4% of the same calves at the age of 26 days, indicating the gradual spread of infection. The isolation rate of Pasteurella multocida did not change much, and varied from 28.6 to 25.7%. No P. haemolytica could be detected. In addition to M. bovis and P. multocida, the herd was also infected with different viruses (including bovine viral diarrhoea virus, infectious bovine rhinotracheitis virus, bovine adenoviruses, parainfluenza-3 virus, and bovine respiratory syncytial virus) as a large proportion of the sera of newborn calves contained colostral antibodies against these viruses. In most of the newborn calves severe clinical signs (fever, depression, inappetence, hyperventilation, dyspnoea, nasal discharge and coughing) due to M. bovis infection developed. The clinical signs appeared already on the fifth day of life, and their incidence was the highest at the age of 10 to 15 days. Three calves (8.6%) died as a result of severe serofibrinous pneumonia. The surviving calves showed very poor weight gain (ranging from 1.5 to 3.5 kg) during the first two weeks of life. PMID- 11402657 TI - The pathological effect of the Bordetella dermonecrotic toxin in mice. AB - The effect of dermonecrotic toxin (DNT) expression of Bordetella bronchiseptica was studied in mice by comparing the pathology induced by a wild type strain with that induced by an isogenic DNT- strain in which part of the structural gene has been replaced by an antibiotic resistance cassette. While extracts of strain B58 proved toxic in intravenously inoculated mice, similar extracts from strain B58GP had lost toxic activity. The parent (B58) and the mutant (B58GP) strains of B. bronchiseptica each possessed comparable virulence for mice. These findings confirmed that DNT production was successfully abolished in strain B58GP while other virulence characteristics required for pathogenicity in mice remained intact, at a comparable level to the parent strain. Turbinate atrophy was observed in mice infected with the DNT+ strain, but not in those infected with the DNT- strain. This indicates that DNT is the cause of turbinate atrophy in the mice and not other factors produced by phase I strains of B. bronchiseptica. B. bronchiseptica DNT showed a lienotoxic effect (lymphocyte depletion and a reduction in the intensity of extramedullar haemocytopoieis) that is considered to adversely alter the immune function of the host animal. In mice infected with strain B58GP, catarrhal pneumonia with characteristic lympho-histiocytic peribronchial and perivascular infiltration was noticed. In mice infected with strain B58, large necrotic areas were seen surrounded by an inflammatory reaction. The DNT appears to directly damage lung tissues, at least in mice. DNT production seems to enhance the establishment of B. bronchiseptica in the lungs, presumably by reducing the local resistance and causing severe local damage to the lung tissues. PMID- 11402658 TI - Epidemiology and characterization of animal Salmonella enterica subspecies Enterica serotype typhimurium DT104 in Hungary. AB - Reports on the internationally emerging significance of multiresistant zoonotic Salmonella in animals and man prompted studies to estimate the significance of multiresistant Salmonella enterica subspecies enterica serotype Typhimurium (S. Typhimurium) phage type DT104 of animal origin in Hungary. A collection of 231 strains (primarily of goose, turkey, poultry and porcine origin from the years 1997-1998) was tested for resistance against 7 selected antibiotics (ampicillin, chloramphenicol, enrofloxacin, nalidixic acid, streptomycin, tetracycline and sulphamethoxazole). Strains with resistance against 3 or more were defined as multiresistant. All strains were phage typed using Felix-Callow's S. Typhimurium phage typing system, and 91 of them (suspect DT104) were also typed according to Anderson's definitive typing (DT) system. In this study, 14% of animal strains from 1997-1998 was classified as DT104, for which turkey, pig and duck seemed to be the main carriers, and the multiresistant non-DT104 strains represented a further 6% of this collection. The prevalence of DT104 was highest among strains of turkey origin (50%), followed by strains of pig (29%), chicken (25%), duck (19%), and goose (3%) origin. The other DT104 related phage types (DT12 and U302) were only detected in the case of 4 strains (2 of porcine, and one each of turkey and of goose origin). The DT104 corresponded to the Felix-Callow types 2/3 or 2c/3 in each case, except in the case of 3 turkey strains where they corresponded to type 35/3. Nalidixic acid resistance was detected in all multiresistant turkey strains and in some of other animal origin but none of these strains were resistant to enrofloxacin. A retrospective analysis (based on the above relationship) indicated that S. Typhimurium strains corresponding to DT104 could be present and increase in the Hungarian farm animal population from about 2% to 20% between 1985 and 1990, in a manner similar to the emergence of human DT104, as reported elsewhere (Paszti et al., 2000). The 91 suspect DT104 strains were also tested for plasmid profile and for spvC gene indicating the presence of the large serotype specific plasmid (Ssp). No characteristic plasmid profile could be attributed to S. Typhimurium DT104. The serovar-specific large plasmid was detected by PCR for spvC in 100% of DT104 strains and in 77% of the non-DT104 strains. The virulence of two DT104 strains was tested in orally infected day-old chicks and compared with virulence of 4 non-DT104 strains. Higher colonizing virulence of DT104 strains could be established as compared to the other strains. PMID- 11402659 TI - Survey on Myxobolus infection of the bleak (Alburnus alburnus L.) in the river Danube and in Lake Balaton. AB - In a three-year survey of myxosporean infections of the bleak (Alburnus alburnus), involving the examination of 205 fish specimens from the River Danube and 50 from Lake Balaton, four Myxobolus species (two gill parasites, one fin parasite and a species parasitising the skeletal muscles) were detected. Two of the species could be identified as M. alburni and M. obesus. Of the other two species, the gill parasite proved to be a hitherto undescribed species which is described here as a new species by the name of M. margitae. One of the two gill parasitic species, M. obesus, formed plasmodia in the respiratory lamellae of the gill filaments, while the plasmodia of M. margitae n. sp. were formed in the afferent artery of the primary gill filaments. The plasmodia containing spores morphologically identifiable with the species M. alburni were located in the connective tissue between the fin rays. The less frequently found muscle parasitic Myxobolus species has not been identified precisely. The plasmodia of M. obesus were found in the fish in May and June, while those of M. alburni and M. margitae n. sp. in July and August. The prevalence of infection in fish examined in these periods was 15.5% for M. obesus, 11.5% for M. margitae and 14.0% for M. alburni. PMID- 11402660 TI - First report on the occurrence of an actinosporean stage (Myxozoa) in oligochaetes from Spanish freshwaters. AB - Oligochaetes living in the Mijares River close to the Sitjar reservoir, and polychaetes from the brackish and marine waters of a channel flowing into the Mediterranean Sea, both in the province of Castellon (Spain), were examined for the presence of actinosporea. An aurantiactinomyxon was isolated from 60 specimens of the oligochaete Branchiura sowerbyi collected from the river, but no actinospores were isolated from 160 polychaetes collected from the sea channel. The aurantiactinospores were detected by the cell-well plate method. The detected species are not identical with any of the aurantiactinomyxon forms hitherto described in the literature. This is the first report on the occurrence of an actinosporea in Spanish waters. PMID- 11402661 TI - Pathological and immunological study of an in ovo complex vaccine against infectious bursal disease. AB - The appearance of very virulent strains of infectious bursal disease (IBD) virus at the end of the 1980s made it necessary to develop more effective immunization procedures. To facilitate this, the immunogenicity and the immunosuppressive effect of a mild (G-87), an intermediate (LIBD) and an intermediate-plus (IBDV 2512) IBDV strain were tested after the in ovo inoculation of 18-day-old SPF and broiler chicken embryos. It was established that no noteworthy difference existed between the immunized and the control embryos in hatching rate and hatching weight. The higher the virulence of the vaccine virus strain, the more severe damage it caused to the lymphocytes of the bursa of Fabricius. In SPF chickens, the haemagglutination inhibition (HI) titres induced by a Newcastle disease (ND) vaccine administered at day old decreased in inverse ratio to the virulence of the IBD vaccine strain, while in broiler chickens this was not observed. Despite the decrease of the HI titre, the level of protection did not decline, or did so only after the use of the 'hot' strain. SPF chickens immunized in ovo with a complex vaccine prepared from strain IBDV 2512 and IBD antibody showed the same protection against Newcastle disease as the broilers. In broiler chicken embryos immunized in ovo, only strain IBDV 2512 induced antibody production, and such chickens were protected against IBD at 3 weeks of age. The complex vaccine administered in ovo has been used successfully at farm hatcheries as well. PMID- 11402662 TI - Studies on the pathogenesis of chicken infectious anaemia virus infection in six week-old SPF chickens. AB - The pathogenesis of chicken infectious anaemia virus (CAV) infection was studied in 6-week-old and one-day-old SPF chickens inoculated intramuscularly with graded doses of Cux-1 strain (10(6)-10(2) TCID50/chicken). Viraemia, virus shedding, development of virus neutralizing (VN) antibodies and CAV distribution in the thymus were studied by virus isolation, polymerase chain reaction (PCR), immunocytochemistry (IP) and in situ hybridization until postinfection day (PID) 28. In 6-week-old chickens infected with high doses of CAV, viraemia and VN antibodies could be detected 4 PID and onward without virus shedding or contact transmission to sentinel birds. However, virus shedding and contact transmission were demonstrated in one-day-old infected chickens. In the 6-week-old groups infected with lower doses, VN antibodies developed by PID 14, transient viraemia and virus shedding were detected. The thymus cortex of all 1-day-old inoculated chickens stained with VP3-specific mAb. Cells with positive in situ hybridization signal were fewer and scattered throughout the thymus tissue of the one-day-old inoculated chickens as compared to IP-positive cells. These results suggest that early immune response induced by high doses of CAV in 6-week-old chickens curtails viral replication and prevents virus shedding. PMID- 11402663 TI - New pig disease in Hungary: postweaning multisystemic wasting syndrome caused by circovirus (short communication). AB - Postweaning multisystemic wasting syndrome (PMWS), a new disease in Hungary, was recognized in a swine herd located in Southeast Hungary, during the early winter of 1999. The first clinical signs of paleness, anaemia, and leanness appeared immediately after weaning, at the age of 40-50 days. Pustules were frequently observed on the skin of the trunk, and signs of necrotic dermatitis were also visible. A syndrome of poor growth and wasting was characteristic of the affected pigs. A porcine circovirus (PCV), the suspected causative agent, was detected by polymerase chain reaction (PCR). Sequencing data and restriction endonuclease (RE) analysis of the PCR products suggested that the virus belonged to the PCV-II group where all the causative agents of PMWS are also grouped. PMID- 11402664 TI - Comparison of the genome of ovine adenovirus types 1 through 5 by restriction enzyme analysis and DNA hybridisation. AB - The DNA of the prototype strains of ovine adenovirus (OAdV) 1 through 5 was analysed by restriction enzyme (RE) digestion. The RE patterns generated by HindIII and PstI enzymes were characteristic of the examined strains. OAdV-2 and 3 resembled each other the most, and their EcoRI and HindIII patterns seemed to be identical. Considering the number of comigrating fragments, serotypes OAdV-2, 3, 4 and 5 looked more closely related to each other than to OAdV-1. This finding was strengthened by Southern blot hybridisations probed with random HindIII clones of OAdV-3. The estimated genome size of the examined OAdV types ranged between 31.9 and 32.8 kilobase pairs. The results supported the new genus classification of OAdVs. PMID- 11402665 TI - Reptile adenoviruses in cattle? AB - This paper describes a hypothesis on the origin of the members of the recently established adenovirus genus, Atadenovirus, invading cattle, sheep, deer, duck and poultry. Comparison of the phylogenetic trees of adenoviruses and their hosts suggests a very ancient but common origin for the atadenoviruses. The surprisingly large difference between these virus types and other adenoviruses infecting the same host can be easily understood by assuming their separate evolution in different hosts (e.g., in reptiles versus a coevolution with mammals and birds, respectively) followed by a later host switch. PMID- 11402666 TI - Porcine adenoviruses: an update on genome analysis and vector development. AB - Although porcine adenoviruses (PAdV) are present in the swine populations worldwide, they usually do not cause any disease, or the infection is only manifested in a mild diarrhoea or respiratory signs. The importance of adenoviruses, however, is constantly growing as there is a possibility of developing them into viral vector vaccines against more significant swine pathogens. A short summary of the well-established facts of porcine adenoviruses is given and recent developments of the genetic analysis of these viruses are discussed in detail. The possibilities of vector development and examples of vector vaccines already reported in the literature are mentioned. PMID- 11402667 TI - Bovine herpesvirus type 4: a special herpesvirus (review article). AB - This paper summarizes the history of and information on bovine herpesvirus type 4 (BoHV-4) from the first isolation to the most recent results. For almost twenty years BoHV-4 has been considered a typical herpes 'orphan' virus, which infects several species but causes no illness. The latest experiments revealed the close relationship of this virus with the immune system and other tissues. The virus was even considered as a possible candidate for a vector vaccine. BoHV-4 as a strange herpesvirus has several features which are not characteristic of other herpesviruses, such as several latency sites, persistence in serum, dividing cells necessary for virus replication, and the wide host range. In addition to describing the main features of the virion, replication, clinical signs, nomenclature problems, this review intends to concentrate on the new and strange results coming out from several laboratories worldwide. It is also suggested that because the virus combines several properties of various herpesvirus subfamilies and because of its close relationship with the immune system, it may deserve further attention as a representative of a potentially new genus within the Gammaherpesvirinae subfamily. PMID- 11402668 TI - Assessment of ovulation of common carp (Cyprinus carpio) females selected for induced spawning on the basis of external morphological characteristics. AB - Secondary sexual characteristics such as softening and rounding of the abdomen as well as reddening and protrusion of the anal papilla and vent can be of help to breeders in selecting common carp (Cyprinus carpio) females prepared for propagation. To assess the reliability of this method, long-term data obtained on induced spawning of common carp at a large-scale fish hatchery were evaluated. The average spawning ratio of 2,620 females receiving hormonal injections was 79.8%. The average pseudogonadosomatic index (PGSI) calculated from data on the egg production of 2,086 females was 16.3 +/- 5.87% (mean +/- SD) for the same period. There was a correlation between fish weight and the time of induction determined by the breeder on the basis of external morphological characteristics. The similarity of the responses of females, including both spawning ratio and PGSI, among the different weight categories proved the reliability of this method for identification. PMID- 11402669 TI - Monoamine oxidase activity in some brain areas of sheep after hormonal stimulation. AB - The influence of hormonal superovulatory preparations Folistiman (450 IU FSH, Spofa, Prague) and serum gonadotropin (1500 IU PMSG, Spofa, Prague) on monoamine oxidase (MAO), the degradative enzyme of catecholamines, was investigated in some areas of the brain regulating reproductive functions (area preoptica of the hypothalamus, pituitary gland, and pineal gland) in ewes with synchronized oestrus (20 mg chlorsuperlutin) during the oestrous period using a radiochemical method. After intramuscular administration of 1500 IU PMSG, marked increase of MAO activity was found in the area preoptica (p < 0.05) and in the pituitary gland (p < 0.01) in comparison with the control group. No change occurred in MAO activity after ovarian stimulation with FSH. Administration of the above superovulatory preparations failed to induce MAO activity in the pineal gland of sheep. PMID- 11402670 TI - Interaction of Dithane M-45 (mancozeb) and lead acetate during a teratogenicity test in rats. AB - The teratogenic effects of lead acetate (Trial 1) and the possible teratogenic effect of this compound administered in combination with a fungicide containing 80% mancozeb (Trial 2) were studied in rats. The test substances were administered by gavage on Days 6-15 of gestation. In Trial 1, five groups were treated with lead acetate administered at doses of 0.1, 0.5, 1.0, 10.0 and 1000.0 mg/kg body weight (bwkg), respectively. In Trial 2, lead acetate was applied at doses of 0.1, 10.0 and 1000.0 mg/bwkg, respectively. In the latter case the dose of the pesticide was 750 mg/bwkg in all treated groups. Lead acetate was not teratogenic after a single administration. Combined administration of lead acetate and mancozeb gave rise to the following toxic effects: average maternal weight decreased during pregnancy, the ratio of live fetuses decreased after the two lowest doses, and fetal mortality increased in the lowest and in the highest dose groups. The ratio of fetal resorption was higher in all the treated groups than in the control group. A significant decrease occurred in average fetal and placental weight in each treated group as compared to the control. Maternal toxicity was expressed in paralysis of the hindlimbs in the two lowest dose groups. Maternal mortality was between 16.7 and 23.3% at the three dose levels. Phocomelia and hernia cerebri occurred as characteristic fetal developmental anomalies in all the treated groups. It is concluded that the joint administration of lead acetate and a mancozeb-containing fungicide can cause maternal toxicity, embryotoxicity and characteristic teratogenic effects. PMID- 11402671 TI - Sequence and expression analyses of the UL37 and UL38 genes of Aujeszky's disease virus. AB - Previously, we sequenced the HSV-1 Ul39-Ul40 homologue genes of Aujeszky's disease virus (ADV), also designated as pseudorabies virus (Kaliman et al., 1994a, b). Now we report the nucleotide sequence of the adjacent DNA that encodes Ul38, the 5'-region (750 bp) of Ul37, and the promoter regions between these divergently arranged two genes. The ADV Ul38 gene encodes a protein of 368 amino acids. Amino acid sequence comparison of ADV Ul38 with that of other herpesviruses revealed significant structural homology. In a transcription study using RNase protection assay and Northern blot hybridization, we found that the Ul38 gene had one initiation site, but the Ul37 gene was initiated at two transcription sites with two potential initiator AUGs, one of which was dominant. Comparison of ADV Ul37, Ul38 and ribonucleotide reductase gene expression showed that these genes belong to the same temporal class with early kinetics. Data of structural and transcriptional studies suggest that regulation of the expression of these two ADV genes could differ from that of the HSV-1 virus. PMID- 11402672 TI - Defence reaction in dental pulp after pulp capping and partial pulpectomy in dogs. AB - The dental pulp was capped indirectly or directly, or partial vital pulpectomy was performed on the 12 functionally most important teeth of 24 beagle dogs. For pulp capping, calcium hydroxide was used, followed by zinc phosphate as a lining, and the preparation was restored with amalgam or composite material. Histological sections were prepared and examined for degree and type of pulp inflammation (hyperaemia, pulpitis, necrosis or gangrene). Degenerative changes of dental pulp (vacuolation, calcification, amyloid or hyaline changes) were also determined. For indirect pulp capping the width of predentine in crown and root dental pulp was measured, and for direct pulp capping and partial pulpectomy (vital pulpotomy) the width and quality of the dentinal bridge were graded. The radical method of partial vital pulpectomy of the coronal part of the dental pulp in dogs produced better quality and continuity of the dentinal bridge than the less radical method of direct pulp capping. This was particularly obvious in small single root teeth such as incisors. PMID- 11402673 TI - Effects of classical swine fever virus infection on the porcine leukocyte subsets. AB - The effects of classical swine fever (CSF) virus infection on the porcine leukocyte subsets were investigated by flow cytometry in acute, chronic and convalescent forms of the disease. The virus antigen could be first detected in the monocytes on postinfection (p.i.) day 10 while in the lymphocytes on p.i. day 13. It could be established that the ratio of CD6+ cells decreased until p.i. day 6, but afterwards it started to increase and reached different values. The CD4+CD8+, the CD8+ and the CD6- cells were obviously higher virus positive than the CD4+ and the CD4-CD8-subsets, but essentially all subsets could be infected. The ratio of CD8+ cells increased during the disease, while the number of double positive cells decreased, and that of the CD4+ cells was variable. The viral antigen could be detected in a lower percentage of the CD4+CD8+, CD8+, CD6+ and CD6- cells of the pigs affected with the chronic form of the disease than in those with the acute form. During the experiments no viral antigen could be detected in the leukocytes of the pig that became convalescent, though the changes in its leukocyte subsets were very similar to those seen in pigs in which the viral antigen could be detected. The studies have revealed that essentially all leukocyte subsets can be infected with the CSF virus, but in very different amounts. PMID- 11402674 TI - Newcastle disease antibody titre is dependent on serum calcium concentration. AB - Chickens were fed diets having optimal, high, and low levels of calcium for 42 days. Serum samples were collected at 14, 28 and 42 days of age, and serum calcium and haemagglutination inhibition titres for Newcastle disease virus were measured. The chickens were vaccinated at 14 days for Newcastle disease. Antibody titres were significantly increased by high dietary calcium and depressed by low dietary calcium. Mean titre was 2.5 (log2) for the optimal diet, 3.2 for the high calcium diet, and 1.6 for the low-calcium diet. Antibody titres were dependent on serum calcium concentration (r2 = 0.98 at 14 days, 0.99 at 28 days, and 0.78 at 42 days). PMID- 11402675 TI - Dog feeding test for assessing the nutritional adequacy of practical diets. AB - The nutritive value of dog foods declared by the manufacturer as nutritionally complete and balanced can be best assessed by feeding trials with dogs. A protocol of a feeding trial has been developed and tested with working dogs fed two different commercial complete and balanced diets for 8 weeks. The parameters used for evaluating the effect of diets were general health status, body and hair coat condition, change of body weight, haematological parameters (white blood cell (WBC) count, red blood cell (RBC) count, haemoglobin, packed cell volume), and biochemical parameters in blood serum (alanine aminotransferase, urea, albumin). The trial protocol proved to be appropriate to monitor the dogs' nutritional status and to reveal differences between diets. This method of evaluation is recommended for use in supporting the nutritional claims (labelling) of dog foods. PMID- 11402676 TI - Cholangiohepatitis in broiler chickens in Japan: histopathological, immunohistochemical and microbiological studies of spontaneous disease. AB - Forty-five broiler carcasses from 6 different flocks were condemned due to liver lesions at processing meat inspection, and collected for pathological and bacterial examinations. All affected chickens showed liver enlargement with discolouration and an apparent acinar pattern. The enlarged gallbladder and the extrahepatic bile ducts contained yellow inspissated cream-coloured material. Histopathologically, extensive proliferation of bile ductules with fibrosis was observed in interlobular connective tissue, and it spread to form bridges with adjoining triads. Destruction and obstruction of portal bile ducts with multiple granulomas due to bacterial infection and outflow of the bile were frequently observed. Many Gram-positive bacilli were seen in the lesions, and they were identified as Clostridium perfringens by indirect immunofluorescence staining technique. Clostridium perfringens was isolated from affected livers. These findings are consistent with cholangiohepatitis. Therefore, it is suggested that C. perfringens might be important in the pathogenesis of cholangiohepatitis in broiler chickens. PMID- 11402677 TI - Effects of saturated and unsaturated fats with vitamin E supplementation on the antioxidant status of broiler chicken tissues. AB - The influence of fish oil (highly unsaturated) and beef tallow (highly saturated) with vitamin E (100 IU/kg) supplementation on the antioxidant status of broiler chicken cockerels was investigated. Chicks were fed a control diet with no added fat, 40 g/kg each of fish oil and beef tallow diets, respectively, from 11 to 42 days of age. Tocopherol concentration and the rate of lipid peroxidation, thiobarbituric acid reactive substance (TBARS) in liver, fatty acid composition of the liver lipids, blood serum total antioxidant status (TAS), and reduced glutathione (GSH) content were determined. Vitamin E supplementation of the diet increased liver alpha-tocopherol content in chicks regardless of the type of dietary fat. Fish oil diet resulted in higher liver TBARS value while beef tallow diet showed lower values compared to the control diet. Vitamin E supplementation reduced liver TBARS as well as serum GSH, and raised serum TAS for all diets. Serum GSH was the same for vitamin E supplemented diets regardless of the fat supplement. Fish oil diets resulted in a significant increase in hepatic lipid n 3 PUFA content. A significant positive correlation was found between liver TBARS and n-3 PUFA content. No relationships were established, however, between liver TBARS and n-6 PUFA or saturated fatty acids. The results suggest that feeding oils rich in n-3 PUFA increases tissue concentration of these fatty acids, consequently increasing tissue lipid peroxidation and reducing the antioxidative status of broiler chickens. Supplementing high levels of vitamin E with such oils may increase tissue oxidative stability. Serum TAS or GSH may be used as a measure of antioxidative status in chickens. PMID- 11402678 TI - Effect of large dietary doses of beta-carotene on plasma retinoid and beta carotene levels and on progesterone production in the granulosa cells of Japanese quail. AB - An experiment was conducted to study the effect of large-dose beta-carotene supplementation on blood retinoid and beta-carotene levels as well as on the progesterone secretion of the granulosa cells in Japanese quail. Laying quails were assigned to three dietary groups. The control group (Group C) received the basal diet (laying feed containing 9000 IU vitamin A/kg). In the treated groups (Groups BC1 and BC2) the basal diet was supplemented with 10(2) and 10(3) mg/kg beta-carotene (BC), respectively. At the end of the two-week feeding period, 10 birds from each group were euthanised. Blood samples were analysed for retinol, retinyl palmitate and beta-carotene concentrations. Granulosa cells were isolated from ovarian follicles (F1 and F2), and PMSG-induced in vitro progesterone (P4) secretion was measured. Similar retinol concentrations were found in both beta carotene supplemented groups, indicating saturation of the retinol-transporting system. beta-carotene supplementation was accompanied by hypercarotenaemia, but did not increase the retinyl palmitate levels in the blood. PMSG-induced P4 production of the granulosa cells decreased significantly in Groups BC1 and BC2 in a dose-dependent manner. PMID- 11402679 TI - Relationships between force-feeding and some physiological parameters in geese bred for fatty liver. AB - The susceptibility of geese of different genotypes and sexes to force-feeding, some plasma biochemical parameters (thyroid hormones, cholesterol, retinoids, total protein and albumin) of force-fed geese, and the relationship between force feeding, fat storage and the above-mentioned parameters were studied. Sixty (30 male and 30 female) geese of three genotypes (Hungarian, Landes and their crossbred called Babat Hybrid) were divided in two groups at 12 weeks of age. Geese in one group (5 males and 5 females from each genotype) received mixed feeding ad libitum. Birds in the other group were force-fed with maize. After 3 weeks all birds were bled, blood samples were taken, and the above-mentioned plasma parameters were determined. Thyroxine (T4) levels were significantly lower in force-fed (11.6 +/- 3.5 ng/ml) than in control geese (22.7 +/- 4.09 ng/ml). Plasma triiodothyronine (T3) level was also lower in the force-fed than in the control group, but the difference was not significant (1.87 +/- 0.23 ng/ml and 2.11 +/- 0.28 ng/ml, respectively). Plasma total protein (TP, 45.2 +/- 4.5 g/l), albumin (ALB, 16.51 +/- 2.8 g/l), beta-carotene (BC, 3504 +/- 3107 micrograms/l), retinol (ROL, 1160 +/- 505 micrograms/l), retinyl palmitate (RP, 1745 +/- 405 micrograms/l) and total cholesterol (TCh, 4.32 +/- 0.55 mmol/l) levels were elevated in the force-fed group as compared to the control (TP = 36.4 +/- 5.1 g/l, ALB = 15.6 +/- 0.9 g/l, BC = 1657 +/- 1681 micrograms/l, ROL = 687 +/- 375 micrograms/l, RP = 1398 +/- 607 micrograms/l, and TCh = 2.83 +/- 1.98 mmol/l). All differences were significant except those found for albumin and beta carotene. No significant sex- or genotype-related effects were observed for these parameters. PMID- 11402680 TI - Gastrointestinal ultrasonography of the dog: a review of 265 cases (1996-1998). AB - The findings of ultrasonography of the gastrointestinal (GI) tract of 265 dogs with GI disorders were analysed retrospectively. The sonographic changes associated with various inflammatory and neoplastic conditions and mechanical obstruction of the GI system were recorded and discussed. Sonographic alterations of the pancreas and the tissues adjacent to the GI tract were also included in the study. Ultrasonographic alterations of the GI tract were classified into three main categories: thickening of the GI wall, changes in peristalsis and dilation of the lumen. Localised thickening of the GI wall with disruption of its structure was caused by both neoplastic diseases and by inflammatory disorders. However, diffuse thickening with retained wall structure was generally associated with inflammatory diseases. The criteria previously established for the ultrasonographic diagnosis of intestinal obstruction were successfully applied to a large number of GI disorders. Pancreatitis was most often associted with hyperchoic mesentery and hypoechoic pancreas mass, but similar alterations were encountered in some cases of gastric or duodenal ulceration. Except in cases of invaginations and intestinal obstructions, the observed ultrasonographic changes were not specific enough for a definitive diagnosis. Nevertheless, ultrasonography proved to be a valuable technique in the diagnostic process of GI disorders of the dog. PMID- 11402681 TI - Influence of electrical stimulation on regeneration of the radial nerve in dogs. AB - The effects of biphasic electric fields on nerve regeneration that follows injury to the left radial nerve were studied in dogs by electromyography (EMG). Left and right radial nerves were crushed with a serrated haemostat. Stimulating electrodes were positioned proximally and distally to the site of the injury. The left nerves received rectangular, biphasic and current pulses (30 microA, 0.5 Hz) through the injury for two months. The right radial nerves were treated as controls and regenerated without electrical stimulation. EMG activities were recorded intramuscularly from the left and right musculus extensor digitalis communis (MEDC). Results obtained at the end of the two-month stimulation period showed a significant difference in EMG activity between the left (stimulated) and the right (non-stimulated) MEDC, suggesting that electrical treatment enhanced nerve regeneration. PMID- 11402682 TI - Questionnaire assessment of the welfare of pigs. AB - A questionnaire study was performed involving 76 farms with a total of 380,207 pigs to assess the welfare of pigs kept in Hungary based on the animal welfare legislation of the European Union. Most significant deficiencies were found in the fulfillment of the provisions relating to space requirements, the stall microclimate and the behavioural needs of the animals. PMID- 11402683 TI - DNA sequence of a small, unidentified plasmid isolated from a Haemophilus somnus strain: short communication. AB - One of the plasmids present in a Haemophilus somnus strain isolated from nasal discharge of a cattle with respiratory disease was purified and cloned for DNA sequencing. The plasmid was found to be 1065 base pairs long with 39.2% G + C content, and showed no homology to any DNA sequenced so far. It has no capacity to code any protein longer than 43 residues. It is not clear yet if this plasmid codes Haemophilus somnus specific factors. PMID- 11402684 TI - The surgical technique and the age of the horse both influence the outcome of mosaicplasty in a cadaver equine stifle model. AB - Six pieces of grafts, 6.5 mm in diameter, 20 mm in length, were taken from each of 170 cadaver hindlimbs, using the cranial surface of the medial femoral trochlea for harvesting. The age of the horses varied between 4 months and 23 years. 30 limbs under the age of 12 years were selected for transplantation. Three of six grafts were transplanted into the medial femoral condyle using different combinations of tunnel depth and dilation. With ageing, a significant decline in transplantability was detected. In general, mosaicplasty cannot be recommended in horses above 11 years. Based on a previous clinical case (Bodo et al., 2000), a good surface alignment was indeed achieved with a combination of graft length drilling and dilation in most cases. However, the occasional entrapment of cartilage debris under the graft prevented perfect alignment in the present cadaver study in 27% of the grafts transplanted in this manner. Since the protrusion of grafts never exceeded 1.5 mm, we conclude that drilling 3-5 mm deeper than graft length with graft length deep dilation can avoid disadvantageous protrusion of the transplanted hyaline cartilage caps, achieving bone decompression at the same time. PMID- 11402685 TI - Pathogenicity of a bovine viral diarrhoea virus strain in pregnant sows: short communication. AB - The biological properties of bovine viral diarrhoea virus (BVDV) strain Oregon C24V were studied after intranasal and subcutaneous infection of pregnant sows. This virus strain is widely used in Hungary for immunising cattle against bovine viral diarrhoea (BVD). Based upon the results of the clinical, gross pathological, histopathological and virological examinations it can be established that the given strain caused asymptomatic infection and serological conversion in sows that were in the second third of gestation. The virus caused clinically apparent disease in some of the piglets born at term, which indicates that it had crossed the placenta. More than half (57%) of the live-born piglets died within 60 days of birth. The sows and their progeny did not shed the virus. BVDV infection has great differential diagnostic importance in pigs, as classical swine fever (CSF) virus strains of reduced virulence cause similar clinical symptoms and gross and histopathological changes. PMID- 11402686 TI - Evaluation of vaccine-induced maternal immunity against classical swine fever. AB - The vaccine-induced maternal immunity against classical swine fever (CSF) was investigated in this study. Eight sows were vaccinated with the Chinese strain of classical swine fever virus (CSFV). The length of time between vaccination and farrowing was 167-217 days. Milk samples from the front, middle and back udder sections and blood samples were taken from the sows on days 3 and 14 after farrowing. Blood samples were obtained from the piglets at the age of 3, 6 and 10 weeks. The antibody level of the milk was examined by ELISA, and that of blood samples by the virus neutralization (VN) test as well. In all 3-week-old piglets and in 80% of the 6-week-old animals the neutralizing antibody level reached the titre of 1:40. In none of the 10-week-old piglets did the titre exceed the value of 1:20, but in about 25% of the piglets it reached 1:20; the half of these piglets came from two litters. In none of the piglets did the antibody level reach the negative threshold in the ELISA test during the study. No significant differences were found between the udder sections in milk antibody level by ELISA. PMID- 11402687 TI - Age- and sex-determined differences in the establishment of tetanus antitoxin production in guinea-pigs. AB - For lack of relevant data of the literature, the tetanus immunisation results obtained in the two sexes were compared in an animal model. Complete immunisation series of weaned, adult and aged guinea-pigs (20-25 animals/group) were performed with aluminium phosphate (AlPO4) adsorbed purified tetanus toxoid (PTAP) as well as with typhoid-tetanus vaccine (TY-TE) containing lipopolysaccharide (LPS). Both vaccines contained 5.0 Lf (limes flocculans, Ramon) per single dose of tetanus toxoid, purity degree: 1500 Lf/mg protein nitrogen (PN). Tetanus antitoxin titres (TAT) were measured after the first shot, and subsequently before and after booster. Compared to TAT of male animals, significantly lower titres were found in female animals after basic immunisation with PTAP in all the three age groups: 1.03 vs. 0.57, 8.75 vs. 5.64, and 0.27 vs. 0.15 IU (international units, related to the Copenhagen International Standard) per ml (sex-chromosome-dependent differences?), as well as in adult animals immunised with TY-TE, before booster: 0.07 vs. 0.02 IU/ml (hormone-dependent differences?). In the latter case the TAT results after booster were 14.49 vs. 12.89 IU/ml. Thus, the lower female prebooster titres were counterbalanced by a quick and effective increase of titres following booster. These results are in accordance with our previous observations in humans (Rethy and Rethy, 1986). From our observations with tetanus immunisation series on guinea-pigs it can be concluded that TAT may be influenced by the effects of sex chromosomes as well as of sexual hormones. During active anti-tetanus immunisation with LPS-containing vaccine (TY-TE) the lower adult female prebooster titres are presumably counterbalanced by the better functionality of the female immune memory. PMID- 11402688 TI - Pharmacokinetics and bioavailability of doxycycline in fasted and nonfasted broiler chickens. AB - The pharmacokinetics and the influence of food on the kinetic profile and bioavailability of doxycycline was studied after a single intravenous (i.v.) and oral dose of 10.0 mg/kg body weight in 7-week-old broiler chickens. Following i.v. administration the drug was rapidly distributed in the body with a distribution half-life of 0.21 +/- 0.01 h. The elimination half-life of 6.78 +/- 0.06 h was relatively long and resulted from both a low total body clearance of 0.139 +/- 0.007 L/h.kg and a large volume of distribution of 1.36 +/- 0.06 L/kg. After oral administration to fasted chickens, the absorption of doxycycline was quite fast and substantial as shown by the absorption half-life of 0.39 +/- 0.03 h, the maximal plasma concentration of 4.47 +/- 0.16 micrograms/mL and the time to reach the Cmax of 1.73 +/- 0.06 h. The distribution and the final elimination of the drug were slower than after i.v. administration. The absolute bioavailability was 73.4 +/- 2.5%. The presence of food in the intestinal tract reduced and extended the absorption (t1/2a = 1.23 +/- 0.21 h; Cmax = 3.07 +/- 0.23 micrograms/mL; tmax = 3.34 +/- 0.21 h). The absolute bioavailability was reduced to 61.1% +/- 4.4%. PMID- 11402689 TI - Mechanism of action of noradrenaline on secretion of progesterone and oxytocin by the bovine corpus luteum in vitro. AB - The present studies were conducted: (1) to determine which beta-adrenoceptor subtypes are involved in progesterone and oxytocin (OT) secretion, (2) to examine whether noradrenaline (NA) acts directly on the cytochrome P-450scc and 3 beta hydroxysteroid dehydrogenase (3 beta-HSD), and (3) to study the effect of prostaglandin F2 alpha (PGF2 alpha) on NA-stimulated steroidogenesis in luteal cells. The effect of NA on progesterone secretion from luteal slices of heifers on days 8-12 of the oestrous cycle was blocked by both atenolol (beta 1 antagonist) and ICI 118.551 hydrochloride (beta 2-antagonist). OT secretion was blocked only after treatment with ICI 118.551 hydrochloride (P < 0.05). Dobutamine (10(-4)-10(-6) M), a selective beta 1 agonist and salbutamol (10(-4) 10(-6) M), a selective beta 2 agonist, both increased progesterone production (P < 0.01) with an efficiency comparable to that produced by NA (P < 0.01). The increase of OT content in luteal slices was observed only after treatment with salbutamol at the dose of 10(-5) M (P < 0.01). Dobutamine had no effect on OT production at any dose. A stimulatory effect of NA on cytochrome P-450scc activity (P < 0.05) was demonstrated using 25-hydroxycholesterol as substrate. 3 beta-HSD activity also increased following NA (P < 0.01) or pregnenolone (P < 0.05) and in tissue treated with pregnenolone together with NA (P < 0.01). PGF decreased progesterone synthesis (P < 0.05) and 3 beta-HSD activity (P < 0.01) in tissue treated with NA. We conclude that NA stimulates progesterone secretion by luteal beta 1- and beta 2-adrenoceptors, while OT secretion is probably mediated only via the beta 2-receptor. NA also increases cytochrome P-450scc and 3 beta HSD activity. PGF inhibits the luteotropic effect of NA on the luteal tissue. PMID- 11402690 TI - Peripheral circulating insulin-like growth factor-I and -II in cattle. AB - Interrelationships between circulating concentrations of the insulin-like growth factors (IGF-I and IGF-II) were investigated in 235 blood samples taken from 145 healthy beef or dairy calves, bulls and cows of different breeds and ages. Autoradiography of Western ligand blots indicated different IGF binding protein (IGFBP) profiles between sera from different categories of cattle. Each IGF radioimmunoassay was validated by determining the effects of IGFBPs, ligand and contraligand, as well as serial dilution and comparison with results obtained after molecular sieve chromatography in acid. In female cattle mean values for IGF-I varied from 5.1 nmol/l in postparturient Holstein cows to 18.5-20.5 nmol/l in growing beef heifers, while mean IGF-II concentrations ranged from 30.0 nmol/l in the cows to 14.7-15.7 nmol/l in the beef heifer calves. In male cattle mean serum IGF-I ranged widely from 8.2 nmol/l in 1-day-old Holstein calves to 67.4 nmol/l in 16-month-old Simmental-type bulls. Mean IGF-II concentrations decreased from 22.9 nmol/l in 1-day-old Holstein bull calves to 11.9 nmol/l in 12-month-old beef bulls. Thus, total molar IGF concentrations were fairly stable in female cattle (24.7-35.1 nmol/l) but extended from 27.3 nmol/l to 81.8 nmol/l in the male cattle. The tendency for a reciprocal relationship between serum concentrations of these growth factors was most obvious in the periparturient cows. PMID- 11402691 TI - Insulin-like growth factor binding proteins and mitogenic activity of partially fractionated sheep amniotic fluid. AB - Amniotic fluid collected from ewes on various days of gestation was examined for the presence of insulin-like growth factor (IGF) binding proteins. IGF-binding proteins with a molecular mass of 40-45 kDa appeared at day 41 of gestation. The level of these major IGF-binding proteins increased during pregnancy and reached a maximum at day 106. Smaller IGF-binding molecules with an approximate molecular mass of 35 kDa and 25 kDa appeared at day 90, also reaching a concentration peak at day 106. The mitogenic activity of sheep amniotic fluid after chromatography on Sephadex G-50 was separated into two peaks. The peak having lower molecular mass corresponded to an elution profile of 125I-IGF-I. The first peak, having higher molecular mass, was eluted immediately after the void volume of column. Electrophoresis and ligand blotting showed that proteins in the first peak had similar properties as IGF-binding proteins. PMID- 11402692 TI - Apoptotic cell death in the porcine endometrium during the oestrous cycle. AB - It has been reported that apoptosis plays an essential role in controlling the physiological cell kinetics in the human and rodent endometrium but this type of death has never been studied in the porcine endometrium. The aim of this study was to investigate the apoptotic cell death in the porcine endometrium during the middle (Days 9-11) and late (Day 13) luteal phase, during the luteolysis (Day 15) and early follicular phase (Days 17-19) of the oestrous cycle. Apoptotic cells were identified by in situ DNA 3'-end labelling method. It was revealed that the greatest number of apoptotic cells in the luminal and glandular epithelium was found on Days 17-19 and on Day 15 of the oestrous cycle, respectively. In the stroma, the greatest number of these cells was found on Days 9-11. Our data have shown that in the porcine endometrium, both epithelial and stromal cells undergo apoptosis and that the number of apoptotic cells varies depending on the phase of the oestrous cycle. PMID- 11402693 TI - Absorption of leucine, alanine and lysine from the rumen. AB - The absorption of three amino acids (leucine, alanine and lysine) from the washed, closed rumen was studied in a short-term (75 min) experiment in situ. The concentration of leucine and alanine did not change in the rumen during the experiment, while that of lysine continuously decreased, and 40% of the total lysine placed in the rumen was absorbed during the experimental period. The rate of absorption decreased in proportion to the fall of amino acid concentration. PMID- 11402694 TI - Radiodiagnostic examination of the swimbladder of some fish species. AB - Radiodiagnostic methods have not been used previously for studying the anatomy and diseases of the swimbladder of freshwater fish species. In this study, the radiographic anatomy of the swimbladder and species-related differences in swimbladder structure were studied on plain radiographs taken of 12 Hungarian fish species of major economic importance. Changes observed by radiography were also studied by conventional parasitological methods. The radiodiagnostic method reported here appears to be a useful complement to diagnostic examinations that have been based merely on dissection so far. It enables evaluation of the pathological lesions in live condition, without causing damage to the fish. PMID- 11402695 TI - Preimplantation genetic diagnosis in cattle: a review. AB - Preimplantation Genetic Diagnosis (PGD) is reviewed and novel fields where it may be applied are investigated. Technical advances of PGD in cattle embryos have already enabled its integration as a part of the MOET (Multiple Ovulation Embryo Transfer) breeding system. PGD for well-defined selection targets can enhance cattle breeding and embryo trade. It allows embryo selection according to their sex, and it may be used to breed special cow lines, or top bulls, by selecting embryos for valuable production traits using Marker Assisted Selection (MAS). A good allelic profile and/or the insertion of a transgene can be detected by PGD. This review article presents the technical requirements for PGD, and shows that this biotechnological method has great economic potential. PMID- 11402696 TI - Determination of residues of pyrethroid and organophosphorous ectoparasiticides in foods of animal origin. AB - Analytical methods were introduced for the determination of residues of ectoparasiticides containing pyrethroid and organophosphate active ingredients in foods. Milk and edible tissues of cows treated with three experimental ectoparasiticides (containing cypermethrin + diazinon, deltamethrin + diazinon and alphamethrin + diazinon, respectively) were assayed for the presence of active ingredient residues. Synthetic pyrethroid residues were not detected in any of the samples processed. Diazinon residues could only be detected in milk samples taken on the first day after treatment (0.005-0.025 mg/kg) and in liver and fat tissue samples taken on the day of slaughtering (0.12 and 0.01 mg/kg, respectively). Permethrin and propetamphos residues were determined in the skin, meat and liver of chickens kept on 'Blotic-B' treated litter and in eggs collected at different times after the treatment of layer houses. Permethrin residues could not be detected in any of the samples (< 0.01 mg/kg). Meat and fat tissues of chickens slaughtered on the day after treatment contained small amounts of propetamphos (0.003 and 0.02 mg/kg, respectively). In the case of chickens kept on the treated litter and slaughtered after one week, active ingredient was not detected in meat, but 0.006 mg/kg propetamphos was present in the fat. The residue content of other samples (liver, egg) was below the detection limit of the applied method at all sampling times. From the food toxicological point of view these pesticide combinations can be used safely if the recommended withdrawal period is observed between ectoparasiticide administration and slaughter. PMID- 11402697 TI - Natural deoxynivalenol (DON) contamination of wheat samples grown in 1998 as determined by high-performance liquid chromatography. AB - A high-performance liquid chromatography--diode array detection (HPLC-DAD) method was developed for determining the deoxynivalenol (DON) content of wheat and other cereals. The samples were extracted with a mixture of acetonitrile and water (84 + 16). Part of the extract was evaporated and purified on Florisil and activated charcoal columns. HPLC separation was performed on a C18 column, using acetonitrile-water (8 + 92) as eluent. Diode array detection (DAD) was performed at 218 and 236 nm, by determination of the UV spectrum. Quantitative analysis was carried out by the external standard method, using the UV spectrum obtained by DAD for confirmation. The recovery rate of DON was 75 +/- 3.1% and the detection limit was 0.05 mg/kg DON. Using this method, the DON content of 99 feeding wheat samples grown in the northeastern part of Hungary in 1998 was determined. Eighty eight percent of the samples originating from three counties contained 0.94 mg/kg DON on the average. The highest individual value was 4.3 mg/kg. DON contamination of wheat was of higher prevalence (100%) and severity (0.27-4.3 mg/kg) in the southeastern county of Bekes than in Szabolcs county located in the northeastern part of Hungary (ratio of positive samples: 82%; DON concentration: 0.05-1.3 mg/kg). The higher than usual DON contamination of feeding wheat can be explained by the rainy summer weather. DON contamination of feeding wheat poses a major risk to the production and animal health status of pig herds. PMID- 11402698 TI - Effects of dietary protein and carbohydrate source on rumen fermentation and nutrient flow in sheep. AB - The effects of decreasing levels of rumen degradable protein (RDP) and nonstructural carbohydrate (NSC) (Diet 1: 74% RDP and 38% NSC; Diet 2: 57% RDP and 32% NSC; Diet 3: 48% RDP and 23% NSC) were studied in cannulated sheep. Total volatile fatty acid (VFA) content rose in response to increasing NSC content. The molar ratio of acetate to propionate was the narrowest for Diet 1. Ruminal concentrations of ammonia and urea increased in response to the rising level of RDP. Flow of organic matter (OM) to the duodenum was increased for Diet 3, which resulted in the lowest apparent and true ruminal digestion of OM. Duodenal flow of total nitrogen (N) increased as RDP content decreased. The highest quantity of undegraded feed protein in duodenal digesta was measured in sheep fed Diet 3. Microbial N flow and microbial efficiency were unaffected by the diets. These results indicate that an NSC level lower than 25% and an RDP content lower than 50% did not exert any negative effect on microbial N production. This phenomenon supports the theory that if the level of RDP is lowered with a concomitant decrease in NSC, uncoupled fermentation cannot be observed. PMID- 11402699 TI - Potential diagnostic test for experimental and natural ovine Taenia hydatigena cysticercosis. AB - An ion-exchange chromatographic fraction of Taenia hydatigena metacestode was evaluated for use in the immunodiagnosis of ovine cysticercosis. Analysis of the fraction by sodium dodecyl sulphate--polyacrylamide gel electrophoresis revealed the presence of a 68 KDa protein. Antibodies against the isolated protein were detected in 7 out of 10 experimentally infected lambs. The diagnostic potential of the 68 KDa protein was further confirmed by testing sera from naturally infected post-mortem positive (PM+) and from apparently healthy groups of animals. Eighty % and 8% of animals were found positive by enzyme-linked immunosorbent assay (ELISA) in the groups of PM+ and apparently non-infected lambs, respectively. PMID- 11402700 TI - A survey of chickens for viable toxoplasms in Croatia. AB - Brain tissues of 716 slaughtered domestic chickens (524 broilers and 192 hens) were bioassayed for viable toxoplasms. Each tissue was homogenized and subcutaneously injected into 4 SPF mice. Six weeks later the mice were euthanatized and their brains microscopically examined for Toxoplasma gondii tissue cysts. Three (0.4%) out of a total of 716 birds were positive. All positive cases were hens. This is the first isolation of T. gondii from chickens in Croatia. PMID- 11402701 TI - Pharmacokinetics, urinary excretion and dosage regimen of diminazene in crossbred calves. AB - The pharmacokinetics, urinary excretion and dosage regimen of diminazene were investigated in crossbred male calves following a single intramuscular dose (3.5 mg x kg-1). Following intramuscular administration, the pharmacokinetics of diminazene was described with a one-compartment open model. The absorption rate constant and absorption half-life were 9.86 +/- 3.06 h-1 and 0.121 +/- 0.40 h, respectively. The value of elimination half-life was 107.5 +/- 8.50 h. The apparent volume of distribution was 0.74 +/- 0.07 L x kg-1. Systemic availability following intramuscular administration was 91.7%. Approximately 65% of the administered dose of diminazene was eliminated in the urine within 24 h of its intramuscular administration. Diminazene was bound to plasma proteins to the extent of approximately 32%. The satisfactory intramuscular dosage regimen of diminazene for calves would be 2.24 mg x kg-1 followed by 1.5 mg x kg-1 at 7 days. PMID- 11402702 TI - Formation of a secondary corpus luteum after ultrasound-guided follicular aspiration in cows. AB - This paper reports the observed formation of a secondary corpus luteum (CL) in the presence of the cyclic corpus luteum, on the ovaries of a cow after ultrasound-guided follicular aspiration for oocyte recovery. The secondary structure, although smaller and lighter (4.97 g vs. 6.02 g) than the natural one, had the typical macroscopic appearance of a corpus luteum. Histological examination of the structure using electron microscopy revealed typical structural features of a natural CL. Mean tissue progesterone concentration was significantly lower in the secondary CL (31.15 +/- 3.11 compared with 58.29 +/- 6.32 micrograms/g tissue of the cyclic CL) and oestradiol-17 beta significantly higher than in the natural CL (108 +/- 11.6 compared with 74.2 +/- 7.81 pg/g tissue). P450scc and P450(17 alpha) mRNA was detected in both structures while P450arom and full-length mRNA FSH receptor were detected only in the secondary structure. PMID- 11402703 TI - Effect of exogenous ovine placental lactogen on basal and prostaglandin stimulated progesterone production by porcine luteal cells. AB - The ability of ovine placental lactogen (oPL) to stimulate progesterone secretion of porcine luteal cells isolated from ovaries in different stages of the oestrous cycle and to support the luteotropic action of PGE2 or to protect the corpus luteum (CL) against the luteolytic action of PGF2 alpha was investigated. oPL in all doses used had no effect on progesterone production of cells isolated from early developing corpora lutea while in doses of 1 and 10 ng/ml it increased oestradiol secretion by this type of cells. In doses of 1, 10 and 100 ng/ml it also increased progesterone secretion of cells isolated from mature corpora lutea in a dose-dependent manner. No influence on progesterone production of cells isolated from regressing corpora lutea was observed. oPL added to the culture media had no effect on PGE2-stimulated progesterone production by cells isolated from mature corpora lutea. However, it exerted a protective effect against the luteolytic action of PGF2 alpha observed in cultures treated with PGF2 alpha alone or in combination with PGE2 in a ratio of 4:1. These studies provide evidence that oPL is luteotropic and supports progesterone production in swine. The fact that oPL acted directly on ovarian steroidogenesis suggests that it may also play some role under non-pregnant physiological conditions. Future studies of structural and functional proteins secreted by the porcine conceptus will help resolve this uncertainty. PMID- 11402704 TI - Studies on the toxic interaction between monensin and tiamulin in rats: toxicity and pathology. AB - The characteristics of the toxic interaction between monensin and tiamulin were investigated in rats. A three-day comparative oral repeated-dose toxicity study was performed in Phase I, when the effects of monensin and tiamulin were studied separately (monensin 10, 30, and 50 mg/kg or tiamulin 40, 120, and 200 mg/kg body weight, respectively). In Phase II, the two compounds were administered simultaneously to study the toxic interaction (monensin 10 mg/kg and tiamulin 40 mg/kg b.w., respectively). Monensin proved to be toxic to rats at doses of 30 and 50 mg/kg. Tiamulin was well tolerated up to the dose of 200 mg/kg. After combined administration, signs of toxicity were seen (including lethality in females). Monensin caused a dose-dependent cardiotoxic effect and vacuolar degeneration of the skeletal muscles in the animals given 50 mg/kg. Both compounds exerted a toxic effect on the liver in high doses. After simultaneous administration of the two compounds, there was a mild effect on the liver (females only), hydropic degeneration of the myocardium and vacuolar degeneration of the skeletal muscles. The alteration seen in the skeletal muscles was more marked than that seen after the administration of 50 mg/kg monensin alone. PMID- 11402705 TI - In vitro ocular irritation toxicity study of some pesticides. AB - The use of animals in toxicological screening is a controversial issue. The Draize eye irritation test receives particular criticism because of the injuries inflicted on the test animals. In recent years various in vitro methods have been developed to replace the heavily criticised Draize rabbit eye test for irritation testing. One of the best-studied alternative methods is the Hen's Egg Test Chorioallantoic Membrane (HET-CAM). In the present studies comparative screening was performed with a set of pesticides to establish parallel data on in vitro (HET-CAM) and in vivo (Draize) results. The tested pesticides included Arelon 500 FW (isoproturon), Banvel 480 (dicamba), Dikamin D (2.4 D), Karathane LC (dinocap), Ronstar (oxadiazon) and Modown 4 F (bifenox). In most cases a good correlation was found between the HET-CAM assessment and results of the Draize rabbit eye test. Although the current form of the HET-CAM test is a valuable prescreen method for predicting the ocular irritation potential of chemicals, and can be used for reducing the number of experimental animals, a number of technical problems must still be addressed before these systems can replace whole animal tests. The HET-CAM test can be a useful component of a battery of tests needed for replacing the Draize rabbit eye test. PMID- 11402706 TI - Gene immunization of mice with plasmid DNA expressing rabies virus glycoprotein. AB - Gene immunization can be an effective vaccine strategy eliciting both humoral and cell-mediated immune responses. We constructed plasmid vectors expressing the full-length Vnukovo-32 rabies virus glycoprotein G under the control of CMV IE promoter and enhancer, adenovirus tripartite leader sequences and poly A signal of SV40. The gene vaccines were evaluated for the ability to elicit neutralizing antibodies and to protect BALB/c mice against lethal rabies virus challenge. First, mice were injected intramuscularly (i.m.) into the left hind leg and by the intradermoplantar (i.d.p.) route with equal amounts of plasmid DNA (0.25-0.1 mg). Two weeks later, immunization was boosted with an additional dose of the DNA. The immunized mice were challenged by intracerebral (i.c.) inoculation of CVS-27 (10-50 LD50) rabies virus. All mice produced anti-rabies virus neutralizing antibodies with a titre of > or = 1:45 after immunization with 0.1 0.4 mg of DNA. In challenge experiments, 83 to 91.6% protection was observed. These results confirm that a DNA vaccine could be a simple and effective solution for preventing the spread of rabies. PMID- 11402707 TI - Immunostimulatory effects of the muramyl dipeptide analogue LK415 in chickens immunized with a vaccine strain of infectious bursal disease virus. AB - The effects of muramyl dipeptide (MDP) synthetic analogue LK415 on the immune response of chickens immunized with a live vaccine against infectious bursal disease (IBD) were studied in two independent trials, using levamisole hydrochloride as comparative immunostimulant. Groups of five-week-old commercial chickens (Isa Brown) were immunized orally with 10 doses of the vaccine strain of IBDV (Winterfield strain). The chickens were then given four injections of the MDP analogue LK415 in a dosage of either 0.25 mg/kg body weight (b.w.) or 2.5 mg/kg b.w. or levamisole at a daily dose of 15 mg/kg b.w. for four consecutive days, starting from the day of immunization. Histological examinations of bursal tissue collected on days 2, 4 and 7 postimmunization (p.i.) showed a lower degree of destruction of bursal follicles and earlier renewal of bursal tissue in LK415 treated chickens compared to levamisole-treated and untreated immunized groups. Compared to the other groups, the LK415-treated chickens showed a significantly higher antibody response to IBDV on days 14 and 28 p.i. (P < 0.01) as measured by commercial ELISA. The present study indicates some potent immunostimulatory effects of the MDP analogue LK415 on the chicken immune system. PMID- 11402708 TI - Characterisation of Rhodococcus equi strains isolated from foals and from immunocompromised human patients. AB - The cultural, morphological, biochemical, serological characteristics and antibiotic susceptibility of 25 Rhodococcus equi strains isolated from lungs and lung abscesses of pneumonic foals and 5 R. equi strains isolated from immuno compromised human patients were examined. All R. equi strains showed common cultural, morphological and biochemical characteristics both with conventional tests and on the basis of their enzyme profile. The R. equi strains examined were resistant to penicillins with the exception of ampicillin, to sulphamethazine and several strains also to sulphamethoxazole-trimethoprim. All strains were susceptible to erythromycin and rifampicin. The strains isolated from humans showed somewhat higher rate of antibiotic resistance to penicillin, cefotaxime, kanamycin, streptomycin, lincomycin, and oxytetracycline. The overwhelming majority (96%) of the equine isolates belonged to serotype 1 in Prescott's serotyping system, while the human isolates could not be serotyped. PMID- 11402709 TI - Abdominal ultrasonography in pet guinea pigs. AB - Ultrasonographic examination of 40 pet guinea pigs was performed in the framework of this study. Preparation of the patient for ultrasonography and the examination procedure are described. Normal ultrasonographic findings of the abdominal organs including the liver, spleen, kidney and urinary bladder are presented and illustrated for the first time in this species. Ultrasonography was found to be a useful investigative tool in the diagnosis of abdominal diseases in guinea pigs. In comparison to other diagnostic methods like physical examination including palpation, haematological examination, radiography and diagnostic laparotomy, ultrasonography offers a rapid, non-invasive and reliable means of diagnosing abdominal diseases in this species. PMID- 11402710 TI - Ultrasonographic detection of abdominal abscess in two guinea pigs. AB - Two guinea pigs (Cavia porcellus) with clinical signs of anorexia, weight loss, depression and abdominal enlargement were examined. During ultrasound examination, a fluid-filled anechogenic structure 3-4 cm in diameter, with echogenic spots and a highly echogenic thick wall, was found in the pelvic region in one case and connected to the liver in the other case. An abscess or a cyst was suspected and surgical treatment including laparotomy was performed. By histopathological examination performed after surgery, a liver abscess was diagnosed in one guinea pig and an abscess in the pelvic region in the other animal. PMID- 11402711 TI - Disease caused by Mycoplasma mycoides subspecies mycoides LC in Hungarian goat herds. AB - The occurrence of a goat disease caused by Mycoplasma mycoides subsp. mycoides LC in Hungary is reported. The disease occurred in two goat herds in the spring of 1999. In one herd 25% of the 4-12 weeks old kids (10 animals) while in the other herd 33% of the 6-12 weeks old kids (20 animals) became affected. The goat kids developed polyarthritis. The most severe lesions developed in the carpal joints. All animals died after 3-8 days of disease. Four dead kids were necropsied. All of them had serofibrinous and purulent polyarthritis, and in two animals bronchopneumonia, fibrinous pleuritis and meningitis were also found. In the articular exudates the presence of mycoplasmas was detected by PCR using a general mycoplasma primer. Mycoplasmas were cultured from the joints of all animals, from the abdominal parenchymal organs of two kids and from the lungs of one animal. The cultured mycoplasmas grew in strikingly large colonies, proved to be glucose positive, arginine negative and phosphatase positive, and liquefied the coagulated serum. They survived incubation at 45 degrees C for more than 24 h. Based upon their biochemical properties, the results of the immunofluorescence (IF) and growth inhibition tests and the sequence analysis of the PCR product, the cultured strains were identified as M. mycoides subsp. mycoides LC. Animals purchased in the previous autumn had been introduced to both farms. The disease may have been introduced with asymptomatic carrier animals, as earlier no similar disease had been observed at either farm. PMID- 11402712 TI - A new, modern, cost-saving micro/macro method for the determination of serum fructosamine. AB - Serum/plasma fructosamine (SeFa) concentration is a reliable indicator used in human diabetic control. Tests for monitoring the carbohydrate/energy metabolism of (farm) animals are less commonly performed in veterinary laboratories, since most of the reliable determinations, both automated and manual, are relatively expensive. The aim of this study was to develop a precise, money- (and time-) saving automated micro method for measuring SeFa. ELISA microplates (20 microL samples and 200 microL reagents) and an automatic microplate autoreader were used. The classical nitroblue tetrazolium (NBT) stain reagent solution of Johnson et al. (1982) was modified using a SIGMA reagent to render it stable for up to one year. SeFa concentrations measured by the new method in 30 human blood plasma samples were compared with values obtained by the standard (generally used) LaRoche kit procedure. Fifteen cow, 13 dog and 18 chicken plasma samples were assayed by the new automated 'micro' method as well as by the manual test tube 'macro' method commonly used earlier. The modified reagent was applied for both methods. The coefficient of correlation (r) between the results obtained by the two methods was consistently between 0.94 and 0.98 (p < 0.001). PMID- 11402713 TI - Hepatic biotransformation profiles of sulphamonomethoxine in food-producing animals and rats in vitro. AB - Hydroxylation and acetylation of sulphamonomethoxine (SMM) and deacetylation of N4-acetyl SMM (N4-AcSMM) were estimated in liver post-mitochondrial supernatants (S-9) from laying hens, female cattle, swine and rats. The formation of hydroxylated SMM, 2,6-dihydroxy SMM (2,6-diOH-SMM), was found only with hen S-9s. N4-acetylation rate of SMM was the highest in pig S-9s, followed by rat, then hen or cow S-9s. All S-9s from the four species deacetylated N4-AcSMM. In hen S-9s, the rate of 2,6-dihydroxylation was higher during incubation at 41 degrees C than at 37 degrees C. PMID- 11402714 TI - Effect of beta-carotene and nucleotide base supplementation on blood composition and immune response in weaned pigs. AB - The effect of synthetic beta-carotene and synthetic nucleotide base on daily weight gain, feed consumption and certain haematological, biochemical and immunological parameters of piglets were studied in a 3-week experiment. Beginning one week prior to weaning, the diet fed to one experimental group of piglets was supplemented with 10% Rovimix Beta-carotene at 875 mg/kg of diet. Synthetic uracil and adenine (98%, Sigma-Aldrich) were mixed into the diet of the other experimental group at doses of 500 mg/kg of diet for each substance. The control group received the basic diet without any supplementation. The changes observed over time in the haematological parameters and in certain biochemical variables could be regarded as physiological. By day 21 of the experiment, beta carotene supplementation had significantly lowered the neutrophilic granulocyte percentage and elevated the lymphocyte percentage, while in the other two groups a change of opposite tendency occurred. At the end of the experimental period there was a decrease in plasma vitamin E concentration due to carotene supplementation (control: 6.1 +/- 1.5, nucleotide: 6.3 +/- 2.5, carotene: 2.3 +/- 1.5 mg/L). Lymphocyte blastogenesis induced by phytohaemagglutinin and concanavalin A increased by 50 and 130%, respectively, in the nucleotide group and by 60 and 30%, respectively, in the carotene group, while it did not change in the control group. The supplements exerted no positive effect on the in vivo cellular immune response. PMID- 11402715 TI - Importance of assisted reproductive technologies in the conservation of wild, rare or indigenous ungulates: review article. AB - Biodiversity is increasingly threatened by intensive agriculture, environmental pollution, extinction of natural habitats and several other factors. Several mammalian species including ungulates have disappeared or are threatened by extinction. However, ungulates play an important role both in the ecosystem and in the economy. In general, species or breeds are considered endangered if their population does not exceed 1,000 individuals. In these cases conservation programmes should be initiated in order to maintain or even increase their number. This review deals with the possibilities and limitations of assisted reproductive technologies (ART) in the conservation of ecologically valuable wild, rare and indigenous ungulates. The methods discussed here are artificial insemination, cryopreservation of semen and embryos, embryo recovery and transfer, in vitro production of embryos, as well as micromanipulation techniques including sperm injection, assisted hatching and cloning. Some of these procedures are already being exploited in the breeding of farm ungulates, but more basic information about the reproductive patterns of wild, rare and indigenous animal species is needed before the routine use of ARTs. PMID- 11402716 TI - Comparative study of different methods for dog semen cryopreservation and testing under clinical conditions. AB - The extenders and freezing rates from three different freezing protocols were combined and compared to each other in order to study the post-thawing acrosome integrity and fertility of frozen dog sperm. A commercial bovine TRIS-base extender (TRILADYL) and two self-made canine semen extenders (Norwegian and Dutch) were combined with a conventional bovine and two canine freezing regimes, and acrosome integrity of frozen/thawed spermatozoa was assessed by fluorescein isothiocyanate conjugated peanut agglutinin staining (FITC-PNA). Differences between freezing/thawing protocols were reflected in the proportion of cells with acrosomal damage and not based on motility results. It was concluded that during dog semen cryopreservation extenders had less influence on the post-thawing sperm quality than did the freezing rates. The optimal extender/freezing rate combination (TRILADYL/Norwegian) was used in the clinical practice to evaluate the fertility of frozen sperm administered by intrauterine insemination using a surgical approach. The pregnancy rate was 57% (4/7), but the average litter size was low (2.8). This may have been due to the insufficient sperm numbers contained in an insemination dose and/or to the incorrect timing of artificial insemination (AI). The final conclusion is that the commercial bovine extender is useful for freezing dog semen, and the TRILADYL/Norwegian freezing protocol is recommended as the most advantageous combination for the freezing of canine semen in the clinical practice. PMID- 11402717 TI - Ram seminal plasma and fertility: results from an ongoing field study. AB - The effects of partial replacement of ram semen diluent with ram seminal plasma on the fertility of ewes were studied. Crossbred Chios ewes (n = 152) were assigned to six groups. The oestrous cycles of the ewes were synchronised at the peak (Groups A, B, C and D) and at the end (Groups E and F) of the breeding season by means of intravaginal sponges impregnated with fluorogestone acetate (FGA) for 14 days. Four hundred IU of PMSG were injected intramuscularly at the time of sponge removal. Ewes of Groups A, C and E were artificially inseminated with ram semen diluted with skim milk extender, while those of Groups B, D and F with ram semen diluted with 50% skim milk and 50% ram seminal plasma. The addition of ram seminal plasma induced a significant increase (P < 0.05) in litter size in Groups B and D when compared with that of Groups A and C (1.85 and 1.88 vs. 1.39 and 1.52, respectively). This increase was not significant when insemination was performed at the end of the breeding season (2.0 vs. 1.4). These results indicate that the addition of seminal plasma can influence the fertility of ewes or the fertilising capacity of extended ram semen to some extent. PMID- 11402718 TI - Arthroscopic autologous osteochondral mosaicplasty for the treatment of subchondral cystic lesion in the medial femoral condyle in a horse. AB - An 11-year-old, Hungarian half-bred stallion was presented with a history of mixed left hindlimb lameness of 6 months duration. Subchondral bone cyst of the medial femoral condyle and injury of the medial meniscus were diagnosed. Osteochondral autograft transplantation (mosaic arthroplasty) was performed, taking grafts from the less weight-bearing medial border of the medial femoral trochlea of the affected limb, and transplanting them into the cyst during arthroscopy. The lameness was evaluated prior to and one year after the operation with a motion analysis system during treadmill exercise. Considerable improvement of the lameness and the clinical signs as well as successful transplantation of the grafts, and a new hard joint cartilage surface of the medial femoral condyle could be detected during follow-up arthroscopy. Osteochondral autograft transplantation seems to bee a possible alternative for treating subchondral cystic lesions of the medial femoral condyle in horses. A new technique for the surgical treatment of a subchondral cystic lesion of the medial femoral condyle in the horse is described. PMID- 11402719 TI - Experimental autologous substitute vascular graft for transplantation surgery. AB - Vascular complications in liver transplantation are a major cause of graft failure and mortality. The aim of the study was to create autologous vascular graft without risk of rejection. Posterior rectus fascia sheath lined with peritoneum was used for iliac artery replacement in seven mongrel dogs. The patency was followed by palpation and Doppler ultrasound. The grafts were removed after one month. Five grafts remained patent. The Doppler showed good, relatively increased flow (median flow rate: 383 cm/sec) after one month in all of the cases. Slight increase in diameter was present in all cases. By microscopy the five patent grafts showed viable morphology, fibroblasts, smooth muscle cells and thin fibrin layer in the wall. The grafts were lined partially with a neoendothelial monolayer and a thin fibrin layer. In conclusion, this graft presents an acceptable patency rate and low thrombogenicity, and could be useful in transplantation. Further investigations are needed to study the effect of immunosuppression and rejection on long-term morphology and patency of the grafts. PMID- 11402720 TI - Studies on the toxic interaction between monensin and tiamulin in rats: effects on P450 activities. AB - Studies were carried out to investigate the effects of monensin and tiamulin, and the simultaneous administration of both compounds on microsomal enzymes in rats. In Phase I of the experiments the effects of monensin and tiamulin were studied separately (monensin 10, 30, and 50 mg/kg or tiamulin 40, 120, and 200 mg/kg body weight, respectively), while in Phase II the two compounds were administered simultaneously (monesin 10 mg/kg and tiamulin 40 mg/kg b.w., respectively). When monensin was administered by itself, it exerted no significant effect on microsomal liver enzymes. In a few cases, slight inhibition of certain enzyme activities was seen. Tiamulin provoked a dose-dependent hepatic enzyme induction. The combined administration of monensin and tiamulin at low doses (10 and 40 mg/kg, respectively) resulted in marked elevation of P450-related enzyme activities. The enzyme induction was more pronounced in females than in males. The results suggest that the simultaneous administration of tiamulin may influence the biotransformation of monensin, possibly increasing the amount of reactive metabolite(s) of the ionophore antibiotic. PMID- 11402721 TI - Effect of immersion fluid temperature on the chicken embryo in teratogenicity tests: short communication. AB - The influence of immersion fluid temperature on the development of the chicken embryo was studied on the day most commonly used for treating incubated eggs in teratological trials. Embryonated eggs were immersed in tap water for 30 min on the 12th day of incubation at 22-25 degrees C or at incubation temperature without a waiting time or after 30 min. The incubation was then continued and the eggs were processed on day 19 of the incubation period. Treatment of eggs at 22 25 degrees C caused a significant increase in embryonic mortality, while the 30 min waiting time did not exert an influence on embryogenesis. PMID- 11402722 TI - [Studies on the chemical constituents of Fissistigma oldhamii]. AB - A morphinandienone alkaloid was isolated from the stems of Fissistigma oldhamii (Hemsl.) Merr. Its structure was identified on the basis of UV, MS, IR, 1HNMR, 13CNMR, DEPT and 13C-1H COSY data as N-methyl-2,3,6-trimethoxyg morphinandien-7 one. PMID- 11402723 TI - [Lignans from Schisandra propinqua (Wall.) Hook. f. et Thoms]. AB - Three dibenzocyclooctadiene lignans were isolated from the stems of Schisandra propinqua (Wall.) Hook.f.et Thoms. Their structures were identified as interiotherin A (I), benzoylisogomisin O (II) and gomisin G (III) by spectroscopic analysis. Compound I was first discovered in Schisandra genus. Compounds II and III were originally isolated from S. propinqua. PMID- 11402724 TI - [Determination of four fractions of Lycium barbarum polysaccharides in different varieties]. AB - The contents of four fractions of Lycium barbarum polysaccharides (LBP) in different varieties were analysed. This method can be used for quality evaluation of Fractus Lycii. PMID- 11402725 TI - [Effect of Cordyceps militaris on the damage of rats induced by n-hexane]. AB - Acutely inhaling n-hexane model was adopted to study the effect of Cordyceps militaris and the damage induced by n-hexane. The results showed that Cordyceps militaris was effective on the resisting lipid peroxidation. The results also showed that acutely inhaling n-hexane could lead to or enhance the damage reaction of oxygen free radicals, which could lead to injure of lipid peroxidation. That may be one of the mechanism of n-hexane toxicity effect on living things. PMID- 11402726 TI - [Effect of shenfukang on nephropathy rats induced by adriamycin]. AB - Adriamycin-induced nephropathy (AIN) model in rats (mainlining 7.5 mg/kg avoirdupois adiramycine) was adopted to explore the effect of Shenfukang (SFK). The results showed that SFK could remarkably reduce the content of urine protein, cholesterol, creatinine and urinary nitrogen in serum, MDA in serum and renal cortex of AIN model rats. And SFK also could increase the content of album and golbulin in serum, the activities of SOD in serum and renal cortex of AIN model rats. CONCLUSION: SFK played an important role on AIN. PMID- 11402727 TI - [Effect of unprepared and prepared "he zi" on contraction of isolated trachea smooth muscle in rabbits]. AB - Isolated trachea smooth muscle in rabbits induced by Ach or KCl was adopted to study the effect of unprepared and prepared "He Zi". The results showed that unprepared "He Zi" couldn't affect the contraction of isolated trachea smooth muscle in rabbit induced by Ach or KCl, and that prepared "He Zi" couldn't affect the contraction of isolated trachea smooth muscle in rabbit induced by KCl. The results also showed that prepared "He Zi" could significantly inhibit the contraction induced by Ach. The inhibition effect may not be associated with cGMP, adrenergic receptor, and release of NO and prostaglandin, and was epithelium independent. Maybe the different content of tannic acid contributed to the different effect between unprepared and prepared "He Zi". PMID- 11402728 TI - [Anti-oxidation effect of water extract of Flos chrysanthemi on heart and brain in vivo and in vitro]. AB - This paper reported the effect of water extract of Flos Chrysanthemi on lipid per oxidation (LPO) of heart and cerebral homogenate from rats incubated in 37 +/- 0.5 degrees C, of heart and cerebral homogenate from mice induced by .OH, of heart and cerebral mitochondria member induced by .OH, which was compared with Radix Salviae Miltiorrhizae. The results showed that the inhibition effect of water extract of Flos Chrysanthemi on above-mentioned LPO was equal to Radix Salviae Miltiorrhizae. It was concluded that water extract of Flos Chrysanthemi could restrain the produce of free radicals and LPO induced by free radicals. PMID- 11402729 TI - [Protective effects of total paeony glycoside on cerebral ischemia mice]. AB - OBJECTIVE: To investigate effects of total paeony glycoside (TPG) on cerebral ischemia mice. METHODS: The models of decapitative mice and acute cerebral ischemia mice caused by ligation of bilateral common carotid arteries were used. RESULTS: TPG could prolong gasp time of decapatative mice, lessen cerebral water content and decrease permeability of cerebral capillary significantly. CONCLUSION: TPG possesses obvious protective effects on mice of acute cerebral ischemia. PMID- 11402730 TI - [Effects of fruits of Hovenia dulcis Thunb on acute alcohol toxicity in mice]. AB - OBJECT: To study the effects of fruits of Hovenia dulcis (FHD) on acute alcohol toxicity in mice. METHOD: Acute alcohol toxicity in mice was used to observe the effects of FHD on sleep time and alcohol concentration in blood. RESULTS: FHD could shorten sleep time distinctly and reduce alcohol concentration in blood of acute alcohol toxicity mice. CONCLUSION: FHD has an obvious effect on acute alcohol toxicity. PMID- 11402731 TI - [Effects of yinchenhao decoction on normal animals and animal models of diabetes mellitus]. AB - OBJECT: To study the effects of yinchenhao Decoction on the blood glucose level in a variety of animal models and its mechanism. METHODS: Normal mice, rats model of ailoxan (ALX)-induced diabetes mellitus (DM), mice model of ALX-DM and rats model of dexamethasone (DX)-induced insulin resistance (IR) were used. RESULTS: yinchenhao Decoction was able to inhibit the hyperglycemia by ALX-induced mice(P < 0.01), significantly reduce level of FBG in normal mice, ALX-DM mice and rats (P < 0.05-0.01), improve IGT and lower 2hBG (P < 0.01) after OGTT in DX-induced insulin-resistant rats. CONCLUSIONS: The results suggest that yinchenhao Decoction possesses similar hypoglycemic action to meftormin and gliclazied. PMID- 11402732 TI - [Study on the inclusions of xindonin A-beta-cyclodextrin and xindonin B-beta cyclodextrin]. AB - The inclusions compounds of Xindonin A-beta-cyclodextrin and Xindonin B-beta cyclodextrin were prepared by the liquid-phase method in aqueous solution. They were confirmed by thin layer chromatography, differential thermal analysis and specific rotation. Changes of ultraviolet spectrum were determined. The composition ratios of inclusion compounds were obtained by the continuous variation method. Solubility of the inclusions were determined. PMID- 11402733 TI - [Primary discussion on the imported materia medica]. AB - The features, with respect to medical use, dosage form, manufacture technology and quality, of licensed imported materia medica and botanical preparation in market were discussed in this paper. The international market competition of Chinese Materia Medica was analyzed, and the countermeasure to resist the imported materia medica was also included. PMID- 11402734 TI - [Pests on zingiberaceous plants and their control in south China]. AB - The pests on the zingiberaceous plants and their occurrence and damage as well as control have been investigated in the ginger plantation of South China Botanical Garden recently. 12 insect pests and 1 mite pest are common. To control them effectively, such as selecting loose, fertile soil as planting field with proper shade, applying healthy and vigorous seeds, seedling and rhizomes for propagation, regulating plant density, sanitizing the field in time are mostly recommended. The application of chemical and biological pesticides and the protection on the beneficial organisms are also stressed. PMID- 11402735 TI - [Wild plants used for the folk dietotherapy in Arhorchin Mongolians]. AB - There are 13 species of native wild plants used for folk dietotherapy by Arhorchin Mongolians in Inner Mongolia. The local people have used those plants as vegetables, fruits and beverages to nourish body, cure scurvy, high blood pressure and 'xira-wusu' disease, repress 'hei' and maintain stomach. PMID- 11402736 TI - [The descriptions and microscopy identification of three allied species of Dysosma versipellis]. AB - The descriptions, histological and microscopical characteristics of three allied species of Dysosma versipellis were described. PMID- 11402737 TI - [The descriptions, microscopical and TLC identification of Scurrula parasitica L. var. graciliflora]. AB - This paper reported the descriptions, microscopical characters and TLC of Scurrula parasitica L. var. graciliflora. PMID- 11402738 TI - [The pharmacognostical identification of peel of Gymnema sylvestre]. AB - This paper reported the descriptions, microscopic UV and TLC identification of peel of Gymnema sylvestre. PMID- 11402739 TI - [Identification of seeds of Cuscuta australis and C. chinensis by TLC and HPLC]. AB - Identification of seeds of Cuscuta australis R. Br. and C. chinensis Lam. was carried out by TLC and HPLC. Polyamide membrane was used as stationary phase, MeOH-HOAc-H2O and CHCl3-MeOH-HOAc were used as mobile phase for TLC. For HPLC, Hypersil-ODS column was used; the mobile phase was MeOH-0.025 M H3PO4; the flow rate was 1.0 ml.min-1; detection wavelength was 360 nm; and column temperature was 40 degrees C. Both methods represented significant identification characteristics, and were simple, accurate and reproducible. PMID- 11402740 TI - [The doctoral degree--devalued or only changed?]. PMID- 11402741 TI - [Stronger centralization of medical laboratories?]. PMID- 11402742 TI - [Supporting belts--prevention gone astray?]. PMID- 11402743 TI - [Treatment of gynecologic cancer]. PMID- 11402744 TI - [Forensic medical aspects of intracranial hemorrhage]. AB - BACKGROUND: Intracranial haemorrhage may be due to pathological processes or trauma even in the absence of a fracture. Since the circumstances surrounding death in such cases are often obscure, the police are notified and a forensic autopsy is performed. MATERIAL AND METHODS: We have retrospectively analysed 116 cases (66 males and 50 females) of intracranial haemorrhages without a concomitant fracture occurring in a series of 4,040 forensic autopsies. RESULT: Intracerebral haemorrhage was most frequently encountered (46%), while subarachnoidal haemorrhage occurred in 38 cases (32.8%), of which only 27 (71%) had aneurysms. In two cases, traumatic subarachnoidal haemorrhage occurred after a fight. Subdural haematomas were found in 24 cases (21%); they were significantly more frequent among men than among women. Intracerebral haemorrhage was often associated with cardiac hypertrophy, while subdural haematomas were strongly associated with alcohol abuse and falls. Intracerebral bleeding in the younger age group was related to arteriovenous malformations, abuse of heroin or amphetamine. In the majority of cases, the haemorrhage occurred at home. In 21 cases there was information about accidents, mainly falls. Eight cases were later brought to court and the perpetrator charged with manslaughter or murder. Hospital admission occurred in 18% of the cases; only half of the patients underwent surgery and died from cerebral complications. INTERPRETATION: This investigation underlines the importance of performing a forensic autopsy in fatalities with head injury or intracranial haemorrhage in order to specify the type of bleeding and possible aetiological factors involved. PMID- 11402745 TI - [Acupuncture contra antiphlogistics in acute lumbago]. AB - BACKGROUND: Acute low back pain is one of the most frequent complaints presented in general practice. This study compares acupuncture and antiphlogistica in the treatment of acute low back pain in general practice. MATERIAL AND METHODS: Among 60 consecutively included patients with acute low back pain, 30 patients were randomized to standardised acupuncture treatment for two weeks, and 30 patients to entero-soluble naproxen 500 mg twice daily for ten days. Effects were observed over six months, and observed for a further 12 months with regard to relapse of low back pain and number of days on sickness leave. RESULTS: There were no differences in pain or stiffness (VAS, physical tests) at inclusion, nor in the reduction of pain or stiffness over a six month evaluation. However, patients receiving acupuncture used significantly less analgetic drugs during the first week after start of treatment than those receiving naproxen (2/28 versus 11/29, p < 0.01). Patients receiving acupuncture also reported fewer new episodes of low back pain (11/28 versus 30/29, p < 0.05) during the 6 + 12 month follow-up. Side effects were frequent in the naproxen group, especially gastro-enteric side effects (0/28 versus 15/29, p < 0.01). INTERPRETATION: Standardised acupuncture treatment seems to be safe and effective in the treatment of acute low back pain in general practice. PMID- 11402746 TI - [Voiding problems and urinary tract complications in adolescents and adults with myelomeningocele]. AB - BACKGROUND: The aim of this study was to describe voiding dysfunction and urinary tract complications in a population above 16 years of age with myelomeningocele. MATERIAL AND METHODS: 51 persons were included in the study. Data were obtained by questionnaires, ultrasound and glomerular filtration rate; in those with intact urinary bladder, by cystometry and videocystography. RESULTS: 30 out of 33 persons with intact urinary bladder were incontinent. Those with daily incontinence described this as a major problem. Cystometry concluded with normal detrusor contractions in three, detrusor hyperreflexia in five, and a detrusor hyporeflexia in 25 persons. Three out of 30 had vesicoureteral reflux. Ultrasound showed mild hydronephrosis and/or scarring in three persons. Average glomerular filtration rate was 86% (50-131%). 11 had Bricker diversion and seven continent reservoirs. 15 out of 18 persons with urinary diversion were satisfied with this solution. In persons with urinary diversion, the average glomerular filtration rate was 78% (44-109%). Ultrasound showed hydronephrosis and/or scarring in seven out of 16. Overall, urinary tract infections last year were reported by 56%, and pyelonephritis was more common in those with urinary diversion. INTERPRETATION: Incontinence is a common problem in adults with myelomeningocele. About one third had upper urinary tract changes, but none had renal failure. PMID- 11402747 TI - [Acute dialysis-dependent renal failure at the Rikshospital in 1998]. AB - BACKGROUND: There are few epidemiological studies on acute renal failure, and as far as we know none have previously been conducted in Norway. The objective of this study was to evaluate the scope, etiology, treatment and results of dialysis dependent acute renal failure based on data from the Norwegian National Hospital in 1998. MATERIAL: This study is based on data from a questionnaire applied to all patients (n = 44) treated with dialysis for acute renal failure at the Norwegian National Hospital in 1998. The questionnaire was designed by a group of Norwegian nephrologists in cooperation with the Norwegian Society of Nephrology. RESULTS: The incidence of dialysis-dependent acute renal failure was 54.4 per million population. Men constituted 75% of the patient population. The most common underlying disorders were heart disease, peripheral vascular disease, and cancer. 90% of patients had oliguric acute renal failure, most frequently due to postoperative complications, septicaemia or drugs, and in most cases the aetiology was multifactorial. Patients received on average nine intermittent hemodialysis treatments and stayed on average ten days in the intensive care unit. 30% of the patients were either heart, lung, liver or bone marrow transplanted (renal transplant patients were excluded). The 60 day mortality rate was 41%. INTERPRETATION: The incidence of dialysis-dependent acute renal failure was relatively high in the Norwegian National Hospital patient pool, combined with a significant mortality rate. These findings are comparable with those of other studies. PMID- 11402748 TI - [Inhalation of a prostacyclin analog (iloprost) in primary and secondary pulmonary hypertension]. AB - BACKGROUND: Vasodilative therapy in the form of calcium channel blockers and, recently, continuous intravenous prostacyclin has improved exercise capacity and reduced mortality in primary pulmonary hypertension. Their clinical value is limited by either low rate of response or serious side effects. These shortcomings could be overcome by the use of iloprost, a stable prostacyclin analogue. Administering it by inhalation, we assessed its short-term efficacy in patients with primary and secondary pulmonary hypertension. MATERIAL AND METHODS: We studied six patients with primary and six with secondary pulmonary hypertension, all with New York Heart Association functional class III or IV symptoms of congestive heart failure. Iloprost was nebulised with 8 l/min of oxygen and administered in increasing doses from 10 to 40 micrograms via a facemask. The haemodynamic effects of iloprost was assessed by right-heart catheterisation. RESULTS: Inhalation of iloprost was well tolerated, and produced a median reduction in mean pulmonary artery pressure from 52 (42-63) to 41 (35 56) mm Hg (p < 0.05). Cardiac output increased from 3.5 (2.8-4.3) to 4.1 (3.1 5.1) l/min (p < 0.05) and pulmonary vascular resistance decreased from 1036 (722 1526) to 753 (446-1107) dyn.sek.cm-5 (p < 0.01). No changes occurred in heart rate, systemic blood pressure or pulmonary wedge pressure. INTERPRETATION: Drug testing with inhalation of iloprost is safe and causes beneficial haemodynamic changes with selective pulmonary vasodilatation. Since the long-term effect of medical intervention is based on the degree of acute pulmonary vascular reactivity, inhalation of iloprost may be a new therapeutic option for severe pulmonary hypertension. PMID- 11402749 TI - [Measurement of gene activity by DNA microarrays]. AB - BACKGROUND: DNA microarray is a tool that can be used to measure in one single analysis simultaneous changes in the activity of tens of thousands of genes. MATERIAL AND METHODS: The method is based upon advanced robotic techniques; High density arrays of DNA probes are placed on a solid surface; this is followed by hybridisation with a fluorescence labelled sample and analysis of fluorescence signals. RESULTS: The analysis create huge data sets which have to be transformed into formats that can be interpreted and correlated with existing knowledge. This means that bioinformatics is an integrated part of microarray analysis. INTERPRETATION: DNA microarray may be used to examine complex physiological and pathological conditions and will most likely be very important in functional studies addressing the structural knowledge of genes obtained through the Human Genome Project. Dedicated microchips are already being tested in the diagnosis of malignant and premalignant diseases and being used to characterize HIV viruses with respect to choice of therapy. PMID- 11402750 TI - [New knowledge derived from measurement of gene expression with the DNA microarray method]. AB - BACKGROUND: The cDNA microarray method offers the first possibility of obtaining a global understanding of biological processes in living organisms, by simultaneous read-outs of tens of thousands of mRNAs. Initial experiments suggest that genes with similar function have similar expression patterns. MATERIAL AND METHODS: Understanding this level of biological complexity will, however, require completely new approaches to data analysis. Computer science methods, such as data mining and knowledge discovery, can synthesize interpretable if-then rules that model the relation between gene expressions and functions and use the rules to classify unknown genes. The huge body of existing biological and medical knowledge makes it necessary to develop methods for extracting knowledge from such repositories. RESULTS: Models of relations between gene expressions and gene functions in a data set from a publicly available source are synthesized semiautomatically and applied to classify unknown genes. Encouraging results have been achieved. The method is applied in the analysis of data from our microarray system which has recently become operational. INTERPRETATION: The principles are of general importance and will be used to evaluate a wide range of complex data sets like decision support in clinical medicine, for situations in which physicians need to handle a large volume of data for each patient. PMID- 11402751 TI - [Fertility preservation in gynecologic cancer]. AB - Recent advances in both the staging and the understanding of the natural history of gynecologic cancers have led to new approaches to treatment. The treatment can now often be tailored to the extent of the disease, and preservation of child bearing ability and/or sexual function may be possible for certain women with early invasive cancers of the ovary, cervix, endometrium, vagina and vulva. Better understanding of the tumour biology and clinicopathologic factors of prognostic significance will allow for individualization of treatment. Management of patients with early gynecologic cancer should be individualized with the risks of conservative therapy balanced against the dangers and advantages of more radical therapy. Specialists in gynecologic oncology and infertility together with an informed patient and her family should make treatment decisions. In this article we present an overview of the therapeutic management of early invasive cancers of the ovary, cervix and endometrium, and present guidelines that may help preserve childbearing capacity. PMID- 11402752 TI - [Role of lymphadenectomy in the surgery of gynecologic cancer]. AB - BACKGROUND: Complete surgical removal of cancer tissue is a goal in modern cancer treatment, and this is one of the arguments for total lymphadenectomy in gynaecologic cancer surgery. MATERIAL AND METHODS: Review of the literature identified through limited searches on Medline, Cochrane, Bibsys and the Internet. RESULTS: Lymphadenectomy improves the staging and represent advanced surgical treatment with considerable complication rates, related to the surgeon's experience. It is likely that lymphadenectomy directly influences survival for vulva and cervical cancer. The effect on survival among endometrial and ovarian cancer patients is unsettled, but prospective randomized trials are ongoing. INTERPRETATION: Correct staging is especially important in randomized trials of treatment alternatives, and it is the basis for individualised treatment. However, the effect of available treatment is not always well documented, hence the mortality and complication rates related to the procedure should be considered in relation to the potential benefit from the relevant treatment in each patient. Special training in the surgical technique is necessary. For rare conditions, this implies centralization. There are promising new approaches like the sentinel node and laparoscopic techniques, and studies of prognostic tumour markers, that may partially replace total lymphadenectomy in the future. PMID- 11402753 TI - [Follow up of urinary tract problems in adults with myelomeningocele]. AB - BACKGROUND: More than 90% of persons with myelomeningocele have a neurogenic bladder disturbance with incontinence and risk of upper urinary tract deterioration. Both aspects need to be considered when planning treatment and follow-up. MATERIAL AND METHODS: The study is based on review of articles and clinical experience. RESULTS: A thorough examination of the patient's voiding methods and incontinence is necessary. Examinations for renal function, reflux and hydroureteronephrosis as well as cystometry should also be carried out. The results of such examinations, together with an assessment of the patient's motor and cognitive function, as well as motivation, will provide a basis for further treatment and follow-up. We suggest a flow-chart for treatment and follow-up of persons above 16 years of age with myelomeningocele. INTERPRETATION: Patients with myelomeningocele should have a thorough examination and an individual plan for treatment and follow-up of their urinary tract dysfunction. Depending on the pathological findings, routine follow-up should be in done intervals from six months to five years. PMID- 11402754 TI - [Routine follow up after treatment for gynecological cancer]. AB - BACKGROUND: Cancer patients are traditionally given follow-up. An earlier diagnosis of recurrent disease is thought to improve chances for curative treatment. The aim of this study was to investigate the approach to routine follow-up after treatment for gynecological cancer. MATERIALS AND METHODS: A questionnaire was sent to all gynaecologic departments in Norway. RESULTS: The number of examinations offered varies from eight to 15 during the first five-year period after primary treatment. Most departments offer four examinations during the first year, two during the second and third years, and one yearly thereafter. There are variations in the routine use of additional tests. Most departments use cytological tests for vulvar, cervical and uterine cancer patients. Chest roentgenography is used more frequently among cervical and uterine cancer patients compared with vulvar and ovarian cancer patients. For ovarian cancer patients, the use of ultrasound investigations is frequent; more than 90% of the departments use blood tests, CA-125 being the most common. INTERPRETATION: There are large variations in follow-up procedures. Advantages and disadvantages with routine examinations are discussed. Most recurrences are detected when they cause symptoms, most often in the interval between visits, and their effect on survival is uncertain. The protocols applied seem to be in line with the pattern of recurrence. Controlled trials investigating alternative protocols in relation to sensitivity, survival, cost and quality of life are important, especially for patients for whom potentially curative treatment is available. PMID- 11402755 TI - [Establishment of the Cancer Registry]. AB - In 1948, the WHO proposed that five countries, Norway among them, should study the incidence of cancer. Three years later the Norwegian Ministry of Health and Social Affairs issued instructions for the reporting of all cases of cancer to the new Cancer Registry of Norway. Its first director was Dr. Einar Pedersen, who was succeeded by Dr. Froydis Langmark in 1983. Between 1957-61 and 1992-96, the number of cancer cases increased from 41,648 to 99,601. Most types of cancer were up; stomach cancer was an exception to this trend. The increase was highest for malignant melanoma and lung cancer, for both types higher among women than among men. There are considerable variations in cancer incidence between Norwegian counties, between the municipalities within the counties, and between urban and rural communities. PMID- 11402756 TI - [A woman with Addison's disease and a bump on the neck]. PMID- 11402757 TI - [Acute adrenal insufficiency--symptoms and treatment]. PMID- 11402758 TI - [Treatment of false croup with racemic adrenaline]. PMID- 11402759 TI - [Physician as a body technician]. AB - "The physician as a technician" is a metaphor for a significant critique of modern medicine. This article approaches some of these critiques. The notion of medicine as an art is sometimes used to remind us of and in praise of the non technical aspects of medicine. After indicating some difficulties with modern conceptions of medicine as art, I bring in the ancient concept of art, "techne", arguing that it avoids some of the difficulties of the modern art concept. Techne actually integrates the scientific, technical and normative aspects of medicine and gives a positive interpretation of the claim that the physician has become a technician. The concept provides medicine with a fruitful framework for understanding both technology and philosophy. PMID- 11402760 TI - [How competent is the physician?]. AB - Medicine's primary concern is ethical; science and technology must be subordinated human concerns. The article argues that ethical wisdom should be considered the most important quality in a doctor, being necessary in order to ensure that the doctor's theoretical knowledge and practical skills are used for the benefit of the patient. A young doctor describes his attempts to make use of philosophical insight--in particular some elements of Aristotle's ethics--while reflecting on his own clinical experiences, trying to counterbalance medicine's "scientific culture". The case is made that knowledge of human nature, respect for the patient's views and values, and acknowledgement of the doctor's fallibility are central to ethical wisdom in medicine. PMID- 11402761 TI - [DNA microarrays and the right not to know]. PMID- 11402762 TI - [Research courses at the faculties--step backward two hundred years?]. PMID- 11402763 TI - [Malaria epidemics in Tanzania]. PMID- 11402765 TI - [The cover picture of the Tidsskriftet nr 7/2001]. PMID- 11402766 TI - [Sjogren's syndrome]. PMID- 11402767 TI - They're back! PMID- 11402768 TI - Comprehensive treatment of traumatic fracture and luxation injuries in the anterior permanent dentition. AB - The increased incidence of traumatic injuries in anterior teeth is a consequence of modern leisure activities. The key factors to successful functional and aesthetic rehabilitation of such injuries are proper diagnosis and adequate primary and subsequent treatment. In severe cases, sequential multidisciplinary treatment is required. All-ceramic crown and post-and-core systems allow for superior aesthetics without compromising function. This article addresses fractures and luxation injuries in the anterior permanent dentition and discusses updated treatment options. The multidisciplinary rehabilitation of a clinical case for optimal function and aesthetics is presented. PMID- 11402769 TI - The one-step versus the two-step impression technique. PMID- 11402770 TI - Application of metal-ceramic technology for anterior fixed prosthodontic restoration. PMID- 11402771 TI - Restoration of enamel and dentin erosion due to gastroesophageal reflux disease: a case report. AB - Gastroesophageal reflux disease (GERD) is a condition where acid contents of the stomach are regurgitated into the oral cavity, which results in continual exposure of the teeth to these acids. Knowledge of the relationship between GERD and dental erosion enables the appropriate diagnosis and treatment of the underlying medical condition as well as the affected teeth. This article details a case report where severe dental erosion was present due to GERD. After management of the disease, treatment (i.e., diagnosis, treatment planning, and restoration) of the eroded dentition is described. PMID- 11402772 TI - Stone models without faces--part II: an international interview. Laboratory perspectives. Interview by Douglas A. Terry. PMID- 11402773 TI - Pulpal healing following acute dental trauma: clinical and radiographic review. AB - Successful treatment of tooth luxations, crown fractures, and root fractures is often compromised by the emergency nature of the injury. Precise radiographic and clinical evaluation is required to facilitate healing and treatment selection. While traditional treatment protocols indicate that dental injuries implying pulpal ischemia require immediate endodontic therapy to avoid healing complications, the clinical investigations discussed herein suggest that a variety of etiologies for pulp survival/pulp necrosis exist. This article discusses the long-term prognosis of pulp necrosis following traumatic dental injuries, and clinical and radiographic criteria for its diagnosis. PMID- 11402774 TI - The use of teledentistry for remote learning applications. PMID- 11402775 TI - Manipulation of the gingival complex to enhance aesthetic treatment. PMID- 11402776 TI - Using clinical practice analysis to improve care. AB - BACKGROUND: Improving clinical outcomes requires that physicians examine and change their clinical practice. Sustaining outcome improvements requires a dedicated and dynamic program of analyzing and improving patient care. In 1992 North Mississippi Health Services (NMHS) implemented a program to improve physicians' clinical efficiency. CLINICAL PRACTICE ANALYSIS ( CPA): CPA uses evidenced-based guidelines and examines each physician's resource utilization, processes, and outcomes for a diagnosis or procedure. Clinical practice profiles are developed, and individual performance is compared to local and national benchmarks and presented to physicians. The CPA process is used on its own or as a component of more comprehensive performance improvements projects. Physicians have been engaged in outcome improvement by more than 55 CPA projects. RESULTS: NHMS has progressively reduced its Medicare loss and its length of stay (LOS) to 4.9 days. Mortality and readmission rates have been reduced in specific diagnoses. The community-acquired pneumonia project reduced the LOS from 7.7 to 5.1 days, decreaesed the mortality rate from 8.9% to 5.0%, and decreased the cost of care from $4,269 to $3,834. The ischemic stroke project reduced the aspiration pneumonia rate from 6.4% to 0% and mortality from 11.0% to 4.6%. Patients' average LOS decreased from 10.7 days to 6.5 days, and their cost of care was reduced by $1,100 per patient. DISCUSSION: Providing individualized data has engaged physicians in improving outcomes. The program has evolved from improving efficiency to managing outcomes and from simple CPA projects to integrated performance improvement projects; however, the CPA process remains the cornerstone of the current process. PMID- 11402777 TI - Applying the domain management model in treating patients with chronic diseases. AB - BACKGROUND: In the health care system in the United States, the management of chronic health conditions and their functional consequences challenge and frustrate patients, caregivers/families, health care providers, and physicians. Contributing factors include a lack of physician and health care provider training and a health system that emphasizes diagnosis and management of acute illnesses. A broader patient care model is required for patients with chronic disease(s). USING THE DOMAIN MANAGEMENT MODEL (DMM) TO CLASSIFY PATIENTS' CLINICAL PROBLEMS: The DMM is a synthesis of approaches used in internal medicine, geriatric medicine, and physical medicine and rehabilitation. All clinical problems, their treatments, and their outcomes can be classified and followed over time in a multiaxial model with four domains-medical/surgical issues, mental status/emotions/coping, physical function, and living environment. APPLICATIONS OF THE DMM IN MEDICAL RECORD TEMPLATES: Use of the four domain headings in standard templates can lead to an improved awareness of all the relevant issues in the management of chronic illnesses. This awareness precedes a physician's implementation of better care processes. Also, good patient care decisions require good information. MANAGEMENT OF FUNCTIONAL PROBLEMS: The DMM can be used to educate care providers and organize care in terms of important and common functional problem (for example, trouble walking, which lacks a standard approach in health care). CONCLUSION: This common framework for the organization, documentation, and communication of patients' care over time will help teach systematic mangement of chronic health conditions and help with future research on complex patient management. PMID- 11402778 TI - Listening to patients and responding with care: a model for teaching communication skills. AB - BACKGROUND: Listening to patients and responding with care requires answering two simple but not so obvious questions: Does the clinician know how to listen? and Does the clinician know how to show patients that he or she cares? The initial task for the clinican is to learn to use listening skills to identify the way in which a patient is most comfortable interacting. Once this is accomplished, the clinician uses this information to move closer to where the pateint is comfortable communicating. MODEL FRAMEWORK: The framework for this model is the continuum of two intersecting axes. The horizontal axis poles are inquisitive and assertive. The vertical axis poles are objective and subjective. People who are most comfortable communicating in an inquisitive way and tend to be objective are called investigators. Those who are most comfortable communicating in an inquisitive way but are subjective in nature are called unifiers. People who are most comfortable communicating in an assertive way and favor subjectivity are called energizers. Those who are most comfortable communicating in an assertive way and tend to be objective are called enterprisers. PUTTING THE MODEL TO WORK: The progression of the typical medical interview, which involves gathering information, making decisions, addressing patient concerns, and discussing long term outcomes requires the clinician to communicate in all four communication modes. Learning to identify how he or she is most comfortable communicating may help the clinician make a smooth transition from one mode to another. PMID- 11402779 TI - Patient-controlled analgesic infusion pumps. AB - Patient-controlled analgesic (PCA) infusion devices allow patients to self administer narcotic analgesics within the limits prescribed by the physician. PCA therapy is typically used for postoperative, obstetric, terminally ill, and trauma patients. PCA pumps deliver solutions intravenously, subcutaneously, or epidurally and allow patient activation by means of a pendant button on a cord connected to the pump or a button directly on the pump. We evaluated nine PCA pumps from six suppliers. Three of these pumps are syringe-type, while the others use cassette-based fluid delivery. Because PCA pumps have often been cited as examples of devices that contribute to medical error (the most significant risk connected with PCA infusion is overmedication), the accident resistance of each device weighed heavily in our testing. The pumps we tested exhibit varying levels of performance, resistance to accidents and tampering, and ease of use. We rate six of them Acceptable. While none of the six units stands out as ideal, they meet most of our criteria, and we consider them somewhat better choices than the rest. We rate one other pump Acceptable (with Conditions) because, in one of its operating modes, it has a drawback that could be dangerous to patients; we consider its use acceptable only if the hospital doesn't employ the operating mode in question. Finally, we rate two pumps Not Recommended because they both have a significant number of disadvantages. PMID- 11402780 TI - Hazard report. I.v. pole clamp on Baxter Flo-Gard 6201 and 6301 infusion pumps- and other brands--may become loose. PMID- 11402781 TI - Focus on infusion pumps. PMID- 11402782 TI - User experience network. Users may inadvertently enter certain Alaris, IVAC, and Siemens multichannel infusion pumps into service mode. PMID- 11402783 TI - Coming of age: Medwire 2001. TMA's new cyber-presence increases its value and visibility. PMID- 11402784 TI - Loss prevention case of the month. Outrageous conduct? PMID- 11402785 TI - The TMA diagnosis and prescription for TennCare. PMID- 11402786 TI - An unusual cause of pelvic mass. AB - BACKGROUND: Pelvic pain with an associated pelvic mass is a common problem in the emergency room (ER) or physician's office. Concerns about ectopic pregnancy, infection, or malignancy usually dominate the diagnostic evaluation. At the same time, domestic violence as the cause of a pelvic mass is seldom suspected by physicians. CASE: A 38-year-old woman came to the ER with left lower quadrant pain and a left pelvic mass. After four hospital days and multiple diagnostic imaging studies, the diagnosis of hematoma caused by physical trauma to the abdomen was elucidated. CONCLUSION: Proper diagnosis of the cause of the pelvic mass could have been made earlier by careful attention to the social history and by recognizing the high incidence of domestic violence as opposed to the relative infrequency of some other diagnostic entities for which the patient was tested. PMID- 11402787 TI - Aspirin in cardiovascular disease. AB - The historic genesis of aspirin as a therapeutic agent is discussed. The use of aspirin as a primary and secondary chemo-preventive agent is reviewed. Those who introduced aspirin in the late 19th century would be surprised by the uses of aspirin today. It is anticipated that existing and further studies on the use of aspirin as a primary preventive agent will establish its utility and a more precise niche for it as a chemo-preventive agent. Reports indicate that it may be useful as a chemo-preventive agent in the prevention and possibly in the treatment of cancer of the colon and other malignancies. This plebeian drug, aspirin, has earned a position, some would say an egalitarian position, along with thrombolysis and primary coronary angioplasty in the emergency treatment of AMI. There are those who would add an intravenous beta blocker and an ACE inhibitor to this regimen. The laborious history of aspirin should again remind clinicians that they should continue to heed the admonition of Alexander Pope when he asserted, "Be not the first by whom the new is tried nor yet the last to lay the old aside." PMID- 11402788 TI - Osteoporosis. PMID- 11402789 TI - Driven to distraction: cellular phones and traffic accidents. PMID- 11402790 TI - Obesity in Canadian children. PMID- 11402791 TI - Obesity in Canadian children. PMID- 11402792 TI - Obesity in Canadian children. PMID- 11402793 TI - D is for drug addiction--and disability. PMID- 11402794 TI - Abuse: a risk factor for low birth weight? A systematic review and meta-analysis. AB - BACKGROUND: Abuse during pregnancy is considered to be a potentially modifiable risk factor for low birth weight (LBW). We conducted a systematic review and meta analysis to determine the strength of association between physical, sexual or emotional abuse during pregnancy and LBW. METHODS: We selected papers for review from an electronic search of MEDLINE (1966-1999), CINAHL (1982-1997) and the Cochrane Library. We retrieved articles using the following MeSH headings and keywords: "infant low birth weight," "fetus," "perinatal care," "pregnancy," "prenatal care," "infant mortality," "violence," "battered women," "spouse abuse," "infant morbidity," "antenatal" and "neonatal." When necessary, we contacted authors to obtain data that were not included in the published material. We analyzed the methodological quality of each eligible study and selected those of the highest quality for meta-analysis. RESULTS: We reviewed 14 studies, of which 8 were selected for meta-analysis. Using a fixed-effects model, we found that women who reported physical, sexual or emotional abuse during pregnancy were more likely than nonabused women to give birth to a baby with LBW (odds ratio 1.4, 95% confidence interval 1.1-1.8). INTERPRETATION: Abuse may be part of a complex interaction of factors that contribute to LBW. PMID- 11402795 TI - Reviewing the reviewers: the quality of reporting in three secondary journals. AB - BACKGROUND: Secondary journals such as ACP Journal Club (ACP), Journal Watch (JW) and Internal Medicine Alert (IMA) have enormous potential to help clinicians remain up to date with medical knowledge. However, for clinicians to evaluate the validity and applicability of new findings, they need information on the study design, methodology and results. METHODS: Beginning with the first issue in March 1997, we selected 50 consecutive summaries of studies addressing therapy or prevention and internal medicine content from each of the ACP, JW and IMA. We evaluated the summaries for completeness of reporting key aspects of study design, methodology and results. RESULTS: All of the summaries in ACP reported study design, as compared with 72% of the summaries in JW and IMA (p < 0.001). In summaries of randomized controlled trials the 3 secondary journals were similar in reporting concealment of patient allocation (none reported this), blinding status of participants (ACP 62%, JW 70% and IMA 70% [p = 0.7]), blinding status of health care providers (ACP 12%, JW 4% and IMA 4% [p = 0.4]) and blinding status of judicial assessors of outcomes (ACP 4%, JW 4% and IMA 0% [p = 0.4]). ACP was the only one to report whether investigators conducted an intention-to treat analysis (in 38% of summaries [p < 0.001]), and it was more likely than the other 2 journals to report the precision of the treatment effect (as a p value or 95% confidence interval) (ACP 100%, JW 0% and IMA 55% [p < 0.001]). INTERPRETATION: Although ACP provided more information on study design, methodology and results, all 3 secondary journals often omitted important information. More complete reporting is necessary for secondary journals to fulfill their potential to help clinicians evaluate the medical literature. PMID- 11402796 TI - The underuse of probiotics by family physicians. PMID- 11402797 TI - Abuse during pregnancy: a quintessential threat to maternal and child health--so when do we start to act? PMID- 11402798 TI - Reports of reports: how good are secondary publications in medicine? PMID- 11402799 TI - Car phones and car crashes: some popular misconceptions. PMID- 11402800 TI - Rationing medical care: rhetoric and reality in the Oregon Health Plan. AB - The Oregon Health Plan (OHP) has been widely heralded as an important innovation in medical care policy and rationing. Oregon's pioneering method of prioritizing funding for health care through systematic and public ranking of medical services has drawn substantial international interest. This paper reviews the experience of the Oregon plan since it began operation in 1994. We argue that widespread misconceptions persist about the significance of the OHP. In particular, there is little evidence that the OHP has operated as a model of explicit rationing. In reality, Oregon has not rationed services, nor has its policy of cutting public coverage for services produced substantial savings. These findings have important implications regarding the desirability and feasibility of adopting a policy of removing items from the list of insured medicare services in Canada. Oregon's experience suggests that drawing the line on medicare coverage would be more difficult and less financially rewarding than advocates claim. PMID- 11402801 TI - Bioethics for clinicians: 26. Assisted reproductive technologies. AB - Assisted reproductive technologies (ARTs) can be very helpful for certain patients, but ethical concerns have been raised about the inherent nature of specific techniques and the contexts in which many techniques are used. Physicians play important roles in supporting those who wish to become parents and in educating patients about impediments to fertilization and way to promote conception. We discuss various ethical issues surrounding ARTs, including family relationships, informed choice, gender issues, embryo status and the commercialization of reproduction, as well as legal and policy issues. We examine the empirical evidence of the effectiveness of ARTs and suggest ways to approach ARTs in practice. PMID- 11402802 TI - Rheumatology: 14. Diagnosis and management of anterior knee pain. AB - A 32-year-old male runner visits his family physician in late spring complaining of pain in his knees over the past month. The pain is mostly anterior but is not well localized. It is aggravated by running, especially on hills, but subsides after the patient stops running. The patient has kept fit during the winter by swimming and occasional cross-country skiing. He has no history of obvious trauma or a noticeable injury. PMID- 11402803 TI - Death but one unintended consequence of gene-therapy trial. PMID- 11402804 TI - The sodium-restricted DASH diet lowers blood pressure. PMID- 11402805 TI - Measles in your office. PMID- 11402806 TI - The pectoralis major myocutaneous flap in head and neck reconstructive surgery: 16 years of experience. AB - Wide defects resulting after resection of malignant tumors of the head and neck need an adeguate closure. In the last 16 years, 85 pectoralis major myocutaneous island flap procedures were carried out for the immediate reconstruction of surgical defects following extirpation of malignant tumors at various sites of the upper aerodigestive tract. The final functional and cosmetic results were satisfactory. Partial necrosis was observed in four cases. We did not have any cases of total flap necrosis. Post-operative fistulas were encountered in 14 cases (surgical closure was not necessary). The application and complications of the pectoralis major myocutaneous flap placed at cervical level are reviewed. The aspects of postoperative swallowing function of such surgery are discussed. Reconstruction with the pectoralis major myocutaneous flap is a safe and versatile procedure, yielding good clinical and functional results in patients with advanced head and neck tumors. PMID- 11402807 TI - [Preliminary data on early discharge after delivery]. AB - Puerpera and baby's early discharging falls within a bigger project that deals with the reduction of medical treatment and with the razionalization of economic resources. We can talk about early discharging when it occurs by 48 hours after a vaginal childbirth and by 96 hours after a cesarean section. When the mother and the baby have been discharged from hospital, their health will chiefly depend on the efficiency of the sanitary organization that works on the territory. If the nursing service at home is competent and specialized, the stay in hospital will get shorter and shorter. The most deepen studies about early discharging after vaginal childbirth agree upon the lack of any risk both for the mother and for the baby. Early discharging is possible without any risk for the mother's health after a cesarean section as well, but only if the woman is well-disposed, if she is at low risk, if she answers to predetermined criteria of eligibility, and if she has an adequate follow-up when she comes back home. On the basis of these evidences, we have traced a clinical outline that draws the early discharging in obstetrics that, in the last five years, has led to a progressive reduction of the stay in hospital both after vaginal childbirth and after cesarean section. PMID- 11402808 TI - [Prevention of preeclampsia with low-dose acetyl salicylic acid: critical assessment]. AB - The Authors present a critical review of the published literature about the effect of low dose of acido acetilsalicilico on prevention and treatment of preeclampic. Beginning from the effects of low daily dose of acido acetilsalicilico on the pregnancy, the Authors present the published datas from 1970 until today, and suggest the present directions for use of acido acetilsalicilico in pregnancy. PMID- 11402809 TI - [High frequency oscillatory ventilation (HFOV) in the newborn. Study of 10 cases]. AB - OBJECTIVE: To evaluate clinical effectiveness of high frequency oscillatory ventilation (HFOV) with a high-volume strategy in severe neonatal respiratory failure from different causes. PATIENTS AND METHODS: Infants with respiratory failure with oxygenation index (O.I.) > 10, independently from weight, gestation age, kind of respiratory disease, previous treatment with conventional mechanical ventilation (IPPV). Patients were treated with respirator Sensormedics 3100 A. Values of O.I. were recorded at start of HFOV and after 1/2, 6 and 24 hours. Also blood gases, arterial blood pressure and vital data were monitored. If HFOV failed infants were allowed to be shifted to IPPV. RESULTS: 10 infants were selected whose gestation age was comprised between 25-38 weeks and birth weight between 640-2620 grams. Mean value of O.I. at the beginning of HFOV was 31.5 +/- 25.1. In all cases but one O.I. decreased rapidly after HFOV and improvement was significant after 6 hours (10.7 +/- 6.3; p < 0.003) still improving after 24 hours (9.5 +/- 5; p < 0.002). The neonate that did not respond to HFOV had severe congenital valvular aortic stenosis and weighted 930 grams. Four infants died: 3 after response of respiratory failure to HFOV and 1 for cardiopathy. Side-effects were: i) edema in all infants that was treated with furosemide, and ii) transient decrease of sistemic blood pressure after start of HFOV in 6 infants that was treated easily with low-dosage dopamine infusion. No significant increase of the rate of complicating disease was observed in survived patients. CONCLUSION: HFOV with high-volume strategy was able to improve rapidly and significantly O.I. in severe neonatal respiratory failure without increasing complications in survivors. Edema was present in all infants and this might depend on the relatively tardy use of HFOV that required higher mean airway pressures. A more precocious intervention has to be considered in clinical practice. PMID- 11402810 TI - Craniofacial fibrous dysplasia associated with primary hyperparathyroidism. AB - Isolated presentation of fibrous dysplasia or primary hyperparathyroidism is a common finding. Only few cases of craniofacial dysplasia associated with hyperparathyroidism have been reported in the literature. A case of a patient with fibrous dysplasia of craniofacial bones associated with primary hyperparathyroidism without additional endocrinophaties or associated disorders, will be presented. Beside the facial dysmorphism caused by dysplasia, the only clinical symptoms were due to the primary hyperparathyroidism. PMID- 11402811 TI - [Cartilage cell stimulation with low-power laser: experimental assessment]. AB - The aim of this study was to verify the effects of laser therapy performed with Ga-Al-As diode laser (780 nm, 2500 mW) on cartilage cells in vitro. The cartilage sample used for biostimulation was taken from the knee of an adult patient. The cultures were divided into four groups: Groups I, II, III were subjected to biostimulation with different laser parameters; Group IV did not received any treatment. The laser biostimulation was conducted for five consecutive days. At the end of the treatment, cell count and MTT tests were performed to assess cell metabolism. The data showed good results in terms of cell viability in the groups treated with laser biostimulation compared to the untreated group. The results obtained with the use of this new low-power diode laser Ga-Al-As device in the biostimulation of the cartilage tissue, permits us to consider the use of this device clinically. PMID- 11402812 TI - [Trauma of the peripheral nervous system: experimental assessments with guided tissue regeneration]. AB - Guided tissue regeneration open interesting perspectives in reconstructive surgery of peripheral nerves. Artificial conduits for nerve repair can be obtained with biodegradable polymers. Lactic and caproic acid copolimers and poliphosphazenes are biocompatible materials and have a slow reabsorption rate. Two types of conduits obtained with Poli[L-lactide-co-6-caprolactone] and poli [bis (etilalanate) phosphazene] were evaluAted as guides for nerve regeneration in an experimental model on two groups of six Wistar rats. Under general anesthesia and with microsurgical technique, the ischiatic nerve was bilaterally isolated. On the right side a segment of the nerve was removed to create a 10 mm gap. The defect was then repaired using the conduit. On the controlateral limb after the creation of the same defect, the nerve continuity was restored using as an autograft the segment removed from the right side. Control were performed at 30, 90, 180 days and consisted in histological and electron microscopy investigations. They showed the gradual degradation of both the conduits without signs of local toxicity. The regeneration of the nerve fibers in the lumen was not significantly different from that observed in the autologous grafts. Both the conduits may be considered effective for guided nerve regeneration, but polyphosphazenes allow the possibility of use the polymer as a carrier for neurite-promoting factors. PMID- 11402813 TI - [Immediate results of surgical treatment of proximal aortic dissection]. AB - From 1984 to 1997, 112 patients have been operated at the Department of Cardiac Surgery of the Azienda Ospedaliera of Parma for acute dissection of ascending aorta. They were 73 males and 39 females of mean age 59.2 years (range 19-79); in the majority they presented a history of essential hypertension and atherosclerotic disease: the first diagnostic suspicion has been confirmed either by angio-CT scan or by CT scan plus aorthography and, from 1994, with transesophageal color doppler echocardiography that is becoming a valuable and precise tool wich can be used in emergency room. Bentall type composite repair or modified Bentall technique has been possible in 37 patients, while in the remaning 75, surgical aortic repair has been achieved by interposition of a dacron vascular prosthesis. Cardiac procedures that were performed in addition were coronary bypass grafthing, resuspension of the aortic valve, AVR. Total arch replacement with reimplantation of brachiocephalic vessels were performed in 6 patients and partial in 10, when intimal tear extended into the arc. Thirthythree in hospital deaths (29.4%) occurred (15 in S.O. and 18 in UCI). Follow up of the hospital survivors was conducted at a mean postoperative interval of 55 months (range 2 to 149 months with 6 late death related to aortic dissection). Our experience shows an increased incidence hypertension and aortic atherosclerosis, compared with marfan's syndrome as a source of acute aortic dissection in the late years, transesophageal echocardiography is a excellent tool for aortic dissection but it may miss dissection localized in other aortic portion, the need of an individualized and conservative surgical approach tailored to the pathology encountered. PMID- 11402814 TI - [Recent myocardial infarction and heart wall rupture: analysis of 21 cases surgically treated]. AB - Left ventricular free wall rupture is a catastrophic complication of acute myocardial infarction and prognosis will depend on the prompt diagnosis by echocardiography, extension of infarct size and prompt surgical treatment. From november 1984, 21 patients (14 M, 7 F, mean age 68.4 +/- 6 years) underwent emergency operation at our Institution. Surgical technique included resection of necrotic or scarred myocardium and repair by Teflon strips-reinforced sutures, or by patching with Dacron or autologous pericardium. Associated procedures were LV aneurysmectomy, CABG, LV endoaneurysmorraphy and mitral valve replacement. Hospital mortality was 33% (7/17 pts). The postoperative course was prolonged in one patient, who needed hemodialysis and in 5 patients who needed IAPB counterpulsation. The mean length of stay in UCI was 3-10 days. Mean follow up is 70.4 +/- 41 months and 3 patients died of non cardiac related death. We conclude that despite high hospital mortality, satisfactory results may be obtained by surgical repairy under emergency condition, with good-long term survival, depending on prompt diagnosis and infarct size extension. PMID- 11402815 TI - Pre-operative evaluation and treatment of patients with chronic venous insufficiency. AB - OBJECTIVE: To describe the use of radionuclide venography (RV) in the detection of the site(s) of blood reflux through incompetent perforating veins (IPVs) of the leg of patients with chronic venous insufficiency (CVI). EXPERIMENTAL DESIGN: Prospective study in which we evaluated our experience in the assessment and treatment of patients with moderate or severe CVI with a follow-up ranging between 6 and 48 months. SETTING: A surgical service of the Department of Surgery of the University of Rome "Tor Vergata". PATIENTS: Fifty-three patients with CVI that were referred to our service to be surgically treated. INTERVENTIONS: Physical examination, RV, and surgical procedures of ligation of IPVs. MEASURES: Based on pre- and post-operative physical examination and RV. RESULTS: The patency of deep venous system and the presence of 1 to 3 leg IPVs was confirmed preoperatively in all the limbs either with physical examination and RV. After surgery, non-ulcerated skin changes improved and all the ulcers healed within 45 days. Postoperative RV showed that the IPVs had disappeared from the affected areas in 56 limbs. Two ulcers recurred after two and six months respectively. CONCLUSIONS: RV showed to be a reliable method to exactly detect IPVs. This method is useful to guide the ligation of IPVs in patients with moderate or severe CVI. PMID- 11402816 TI - [Characterization of the HUMTPOX and HUMCSF1PO polymorphic systems in a sample of the Parma population]. AB - A population study in a sample from the Province of Parma was carried out to investigate the polymorphic systems HUMTPOX and HUMCSF1PO. Separation of duplex PCR co-amplified fragments was performed by denaturing PAGE followed by silver staining; seven and six allelic variants respectively were found and frequency distribution was in good agreement with previous studies on Caucasian. The power of Discrimination (PD) was 0.82 and 0.88 respectively and the chance of exclusion (MEC) 0.401 and 0.502; the robustness of the polymorphic systems and informative strength, though not particularly high, suggest the use of these polymorphisms in forensic investigations. PMID- 11402817 TI - [Malformative giant emphysema bulla. Report of a case in childhood]. AB - Bullous pneumopathy is considered a surgical disease, although her treatment is still discussed. The initial attempts of surgical treatment were founded on erroneous physiopathological concepts and date back to early years of 1900. These surgical treatments intended to external drainage of giant bullae, their marsupialized or to reduction of extension of bullae by pneumoperitonaeum, section of phrenic nerve or thoracoplasty. The definition of emphysematous bullae has been much improved with the development of computed tomography. A precise study of their size, of their position and of condition of residual pulmonary parenchyma may be considered important to decide the surgical treatment: this must determine the removal of bulla and the reexpansion of compressed pulmonary parenchyma. The Authors report a case of giant bulla, initially wrong interpreted as pneumothorax, of exceptional observation for her malformative origin, diagnoses and surgical treated on an eleven years old child. PMID- 11402818 TI - [One-lobe Caroli's disease (report of a clinical case]. AB - A patient affected by Caroli's disease without congenital fibrosis, 80 y. old is reported. The Caroli's disease was asymptomatic all life long; this was a intraoperative finding and only in the last year the patient presented recurrent cholangitis. The "simple" Caroli's disease is less common than other forms. We analyze the literature and the ethiopathogenetic mechanisms hypotisized. PMID- 11402819 TI - [Prevention of perinatal infection caused by group B beta-hemolytic streptococcus]. AB - Streptococcus agalactiae strains or group B streptococci (GBS) are the leading cause of bacterial pneumoniae, sepsis and meningitis in neonates. GBS is also a major cause of bacteriemia in pregnant women. Colonization of the human rectovaginal tract with GBS is a risk factor associated with chorioamnionitis and transmission of the infection to the infant. Neonatal exposure to high concentrations of GBS, mainly during vaginal delivery, leads to colonisation of the lung airways and subsequent onset of severe diseases like pneumonia, sepsis and menigitis. GBS is present in the genitourinary tract of 10% to 40% of pregnant women, about 50% of the newborns of these mothers will be colonised during delivery and of these neonates, 1% to 2% present a severe invasive disease. The early-onset disease, appear in the neonates within 7 days of life and more than 90% occur within the first day of life. Fatal infection is associated commonly with fulminat and overwhelming early-onset disease. Maternal intrapartum chemoprophylaxis is able to prevent the transmission of GBS to the newborn and to reduce the frequency and the severity of early onset disease. In many countries, in particular in US, several recommendations have been proposed to prevent the perinatal GBS infection. In this paper some recommendations to prevent GBS disease of the newborn, performed in collaboration with Italian Society of Perinatal Medicine, are presented. The most important problem in the prevention programme is the identification of the cases to treat, since it is not possible to give antibiotics to all the women. We combine two strategies for the identification of the women to be treated, one risk based and the other screening based. Intra-partum administration of ampicillin or penicillin is recommended for the women with one or more risk-factors (labour < 37 weeks of gestation, duration of ruptured membranes > = 18 hours, intrapartum temperature > = 38 degrees C, previous infant with invasive GBS disease, diabetes) and for women with collect vaginal and rectal swab for GBS culture at 36-38 weeks' gestation, positive for GBS. No treatment is required for the babies of women intrapartum treated or with negative culture performed near term. Treatment with ampicillin is necessary, only in the new-borns of women with incomplete or unknown results or not done cultures and in those born from mothers with positive cultures, but not intrapartum treated. Collection of swabs for GBS is recommended before antibiotic administration. If the culture is negative, we suggest to stop the antibiotic therapy, otherwise the treatment must be continuated for 5-7 days. In conclusion, a written protocol for prevention of GBS infection in new-born must be adopted in every delivery centre and one possible protocol is proposed in this paper. PMID- 11402820 TI - [Allelic distribution of the HUMCD4 polymorphic system in the Province of Parma]. AB - Allele frequencies of polymorphic system HUMCD4 were determined in a sample from the Province of Parma. Good separation of PCR amplified fragments was performed by non denaturing PAGE; seven allelic variants were found and frequency distribution was in good agreement with previous studies on Caucasian. The statistic tests (Hardy-Weinberg equilibrium -H-W-., Power of Discrimination -PD- and Mean Exclusion Chance -MEC-) and the robustness of the technique suggest the use of this polymorphism in forensic investigations. PMID- 11402821 TI - I did this to you, Mrs. Jones? PMID- 11402822 TI - Computerizing your operatory ... and living to tell about it. PMID- 11402823 TI - Simple diagnosis and treatment planning of the implant patient. PMID- 11402824 TI - [Using high resolution sectional imaging in diagnosis of the eye and orbit]. PMID- 11402825 TI - [Papillary hemodynamics in patients with normal pressure glaucoma and hemorrhage of the optic papilla circumference]. AB - BACKGROUND: Optic disc hemorrhages in patients with normal-pressure glaucoma (NPG) are usually regarded as a sign of vascular dysfunction and as an indicator for glaucoma damage progression. METHODS: Optic nerve head blood flow was measured in 21 patients suffering from NPG with acute optic disc hemorrhages by scanning laser Doppler flowmetry at various locations of the optic disc. Intraocular pressure and mean deviation of the visual field were also monitored. Two groups served as control: 21 patients with NPG matched for age, sex, and stage of the disease and in addition the contralateral eye without any hemorrhages. RESULTS: Optic nerve head blood flow as a mean of several locations was significantly lower in eyes with optic disc hemorrhages than in controls and differed significantly from the contralateral eye. CONCLUSION: Optic nerve blood flow was lower in NPG eyes with optic disc hemorrhages than in the contralateral eye and in controls. PMID- 11402826 TI - [Ocular hemodynamics and visual field in glaucoma treated with dorzolamide]. AB - OBJECTIVE: It has been shown that oral carbonic anhydrase inhibitors improve visual function in glaucoma. Furthermore topical dorzolamide might improve ocular hemodynamics, as was demonstrated previously. This study was undertaken to evaluate whether topical dorzolamide affects visual function and ocular hemodynamics in glaucoma. METHODS: In a retrospective, open clinical trial, dorzolamide eye drops were administered to 28 patients with confirmed primary open angle glaucoma (POAG) in both eyes, 3 times daily for a mean follow up of 9 months. One eye was randomly chosen for evaluation. IOP, blood pressure, heart rate, pulsatile ocular blood flow (POBF) and Humphrey 30-2 visual fields were measured at baseline and after the start of the therapy. POBF was determined by pneumotonography. For statistical analysis the Wilcoxon-matched-paired test and the Bonferoni-Holm adjustment were used. RESULTS: In dorzolamide-treated patients the IOP dropped from 18 mmHg to 15.5 mmHg after 9 months therapy (p < 0.01) and the visual field improved significantly by 18% (p < 0.05). A statistically significant change was found for POBF from 543 microliters/min to 675 microliters/min (p < 0.05). CONCLUSIONS: The results showed the expected drop in intraocular pressure. Visual function and pulsatile ocular blood flow improved significantly which might be explained by an analogous, vasodilatory effect as was observed in orally applied carbonic anhydrase inhibitors. PMID- 11402827 TI - [Analysis of the aqueous humor in keratoplasty patients with keratitis. Initial results]. AB - OBJECTIVE: Herpetic keratitis is a common indication for corneal transplantation. In this patient group especially, there is a relatively high risk of graft failure, partly because of viral recurrence. It can be difficult to clinically distinguish stromal herpetic recurrence from early endothelial allograft rejection. Also a perioperative observation of viral activity seems advisable because of therapeutic consequences. For these reasons we use aqueous humor analysis in certain corneal transplant patients to determine intraocular antibody production. The aim of this study was to evaluate the diagnostic value of such an analysis of aqueous humor. MATERIAL AND METHODS: A total of 28 samples of aqueous humor were obtained from 24 eyes and all samples were tested for antibodies against herpes simplex virus (HSV), most samples (26/28) were tested for antibodies against varizella zoster virus (VZV) and some samples (6/28) for antibodies against cytomegalovirus. We used a modified micro-ELISA technique to detect intraocular IgG production. RESULTS: In 14 samples (50%) we found antibodies against HSV, in 7 samples (25%) against HSV and VZV, in 1 sample (3.6%) against VZV and 6 samples (21.4%) were negative for all antibodies tested. CONCLUSION: The results of aqueous humor analysis led to a specific local or systemic antiviral therapy perioperatively or in the case of postoperative recurrence of herpetic keratitis in most patients. Some patients could be spared long-term treatment with antiviral agents because of negative results in the aqueous humor analysis. PMID- 11402828 TI - [Screening of myopic LASIK patients with increased epithelial wound healing]. AB - BACKGROUND: Wound healing of the cornea is critical for the refractive outcome of myopic laser-assisted in situ keratomileusis (LASIK). As epidermal growth factor (EGF) is important for the origin of epithelial hyperplasia, this study examined preoperative EGF mRNA concentrations in the corneal epithelial cells to detect patients with increased epithelial wound healing response. PATIENTS AND METHODS: The epithelium was biopsied before LASIK in 35 eyes with myopia of -10.0 D. The EGF mRNA concentration in the epithelial cells was quantified by polymerase chain reaction and enzyme-linked oligosorbent assay, and the correlation with postoperative refraction at 6 months was assessed. RESULTS: All eyes were around emmetropia 3 weeks after the surgery. At 6 months postoperatively 27 eyes were within +/- 1.0 D of emmetropia while 8 showed regression of 2.0-4.0 D. Higher EGF mRNA levels were found in eyes with regression than in eyes with postoperative emmetropia. CONCLUSIONS: Preoperative EGF mRNA concentration in the corneal epithelial cells may be an indicator of postoperative refractive outcome of myopic LASIK and offers a new possibility for pharmaceutical manipulation. PMID- 11402829 TI - [Virus relevant physicochemical parameters of viscoelastics in cataract operation]. AB - PURPOSE: To define the physicochemical parameters of viscoelastics that are most important for visual acuity after phacoemulsification. MATERIAL AND METHODS: Eleven commercially available viscoelastics (ophthalmic viscosurgical device, OVD) were used in consecutive cataract operations. In addition to clinical findings, we assessed visual acuity, intraocular pressure, endothelial cell density, and pachymetric data preoperatively and on the first postoperative day. RESULTS: On the first postoperative day the loss of endothelial cells in all phacoemulsifications was 0.661%, corrected for 1 s phacotime, while corneal thickness increased by 0.2384% and visual acuity by 5.206%, and intraocular pressure decreased by 0.306%. The most important parameters for visual acuity were the concentration of sodium hyaluronate and viscosity (100/s.) of the OVD. Pachymetric data were more relevant for postoperative visual acuity than endothelial cell counts. CONCLUSION: Early visual rehabilitation after phacoemulsification depends on stable pachymetric data. OVD with a high content of sodium hyaluronate and high viscosity significantly reduces corneal swelling and improves visual acuity after phacoemulsifications on the first postoperative day. PMID- 11402830 TI - [Results of the Hughes operation]. AB - BACKGROUND: There are several methods for the repair of large defects of the upper and lower eyelids, but we prefer the original procedure by Hughes. This operation has been used traditionally for partial and total reconstruction of lid defects after tumour resection in 7-10%. The advantage of this technique is the rebuilding of the eyelid with the lid tissue itself. We present the results of operations by the Hughes method over a 10-year-period. PATIENTS AND METHODS: Between 1986 and 1996 we operated on 60 out of 600 patients with neoplastic tumours of the eyelids and 1 patient with a large traumatic defect of the upper eyelid by the original operation method of Hughes. We report on a retrospective study of 55 patients including 30 males and 25 females with a mean age of 68.6 years. The duration of the histories of the tumours ranged from 2 month to 10 years. The lower eyelids were involved in 46 patients and the upper lids in 9. In 80% (45 patients) we found basal cell carcinomas and in 17% (9 patients) squamous cell carcinomas. RESULTS: Therapy of tumours: 50 out of 54 tumours (92.6%) were resected in sano and 4 not. During the dissection of the lids we carried out a tissue biopsy but no further tumour tissue was found in three patients. The fourth patient needed three further operations before recovery. In six patients we recognised a new tumour, one of them suffered from Lewandowsky-Lutz-Syndrom, developed a new tumour in the orbit and required exenteration orbitae. Early complications: arterial bleeding in one case, in four cases mild dehiscence of the wound between the graft and the remaining lid, in two patients with high risk, infection of the wound, one rejection of the skin graft, one small infiltration of the cornea. Late complications: In six cases we recognised recurrence of the tumour and three patients with entropium and seven patients with heavy trichiasis needed revision. Minor complications such as mild distichiasis, partial incomplete closing of the lids, a smaller lid fissure or notching of the canthus of the lid which did not require revision. The results were quite satisfactory from a functional and aesthetic point of view. CONCLUSIONS: We recommend the operation method of Hughes for reconstruction of large defects of the eyelids of any reason. PMID- 11402831 TI - [Vision and perception in traffic. Contribution of psychological traffic diagnosis and the section of ophthalmology]. PMID- 11402832 TI - [Scanning microscopy and immunohistochemical studies of an explanted phacic posterior chamber lens (ICL-M, STAAR)]. AB - OBJECTIVE: This study was performed to find out if an explanted phacic collamer IOL (ICL/STAAR) showed structural or surface quality changes after an intraocular implantation. MATERIAL AND METHODS: We explanted an ICL 6 months after implantation and compared it to a new reference lens using scanning electron microscopy and immunohistochemical investigations. RESULTS: The explanted ICL showed no changes in comparison to a new ICL except for a coating with an unidentified material. Both the new and explanted ICL presented grooves on the surface and superficial holes of unknown origin measuring roughly 10 mu. Neither pigment nor macrophages could be found on the explanted lens. Although the lens material contained 0.1% collagen, the immunohistochemical staining for collagen was negative. CONCLUSIONS: The collagen content of the ICL could not be proven using immunohistochemistry. No changes were found between the new and the explanted lens, except for a coating of the explanted lens with an unidentified material. PMID- 11402833 TI - [Keratopathy as a sign of multifocal congenital sensory polyneuropathy. A case report]. AB - PATIENT: A 63-year-old female with bilateral recurrent corneal ulcerations for 10 years, suffered from vascularisation of the cornea and absence of corneal sensitivity. Other symptoms were multifocal hypoaesthesia with hypalgesia of hands and legs, generalised lack of deep tendon reflexes, absence of somato sensory evoked potentials (SSEP) and of sensory nerve action potentials (SNAP) in these regions. A sural biopsy demonstrated extreme lack of myelinated fibres. Acquired causes for polyneuropathy were excluded. THERAPY: Subsequent to local ocular treatment we carried out a perforating corneal transplantation of the left eye because of corneal scars. This had to be repeated 2 years later because of vascularisation of the transplant. The visual function of the left eye could be stabilised at values between 0.2 and 1/50. CONCLUSION: Anamnesis and clinical symptoms of the patient are compatible with the diagnosis of hereditary sensory neuropathy type II (HSN II) affecting the trigeminal nerves. In patients with neuropathy and impaired corneal sensitivity, a favourable prognosis may be achieved by a corneal transplant. PMID- 11402834 TI - [Bilateral vision loss during a stay in tropical surroundings. Bilateral choroiditis in stage II syphilis ]. PMID- 11402835 TI - [Eyelid edema and somnolence after hay fork injury of the eye. Penetrating orbital and brain injury]. PMID- 11402836 TI - [Intraocular (oculocerebral) non-Hodgkin lymphoma]. PMID- 11402837 TI - Status and problems of fusion reactor development. AB - Thermonuclear fusion of deuterium and tritium constitutes an enormous potential for a safe, environmentally compatible and sustainable energy supply. The fuel source is practically inexhaustible. Further, the safety prospects of a fusion reactor are quite favourable due to the inherently self-limiting fusion process, the limited radiologic toxicity and the passive cooling property. Among a small number of approaches, the concept of toroidal magnetic confinement of fusion plasmas has achieved most impressive scientific and technical progress towards energy release by thermonuclear burn of deuterium-tritium fuels. The status of thermonuclear fusion research activity world-wide is reviewed and present solutions to the complicated physical and technological problems are presented. These problems comprise plasma heating, confinement and exhaust of energy and particles, plasma stability, alpha particle heating, fusion reactor materials, reactor safety and environmental compatibility. The results and the high scientific level of this international research activity provide a sound basis for the realisation of the International Thermonuclear Experimental Reactor (ITER), whose goal is to demonstrate the scientific and technological feasibility of a fusion energy source for peaceful purposes. PMID- 11402838 TI - Differentially expressed regulatory genes in honey bee caste development. AB - In the honey bee, an eminently fertile queen with up to 200 ovarioles per ovary monopolizes colony level reproduction. In contrast, worker bees have only few ovarioles and are essentially sterile. This phenotype divergence is a result of caste-specifically modulated juvenile hormone and ecdysteroid titers in larval development. In this study we employed a differential-display reverse transcription (DDRT)-PCR protocol to detect ecdysteroid-regulated gene expression during a critical phase of caste development. We identified a Ftz-F1 homolog and a Cut-like transcript. Ftz-F1 could be a putative element of the metamorphic ecdysone response cascade of bees, whereas Cut-like proteins are described as transcription factors involved in maintaining cellular differentiation states. The downregulation of both factors can be interpreted as steps in the metamorphic degradation of ovarioles in worker-bee ovaries. PMID- 11402839 TI - Spider silk has an ice nucleation activity. AB - Several ice nucleating substances have been identified, which exist in vivo or can be extracted from biological materials. Spider silk, which has a strong ability for water condensation, has also been found to possess an ice nucleation activity. The freezing temperature of water droplets was higher in the presence than in the absence of spider silk. Moreover, by means of environmental scanning electron microscopy, it was observed that the activity is not due to foreign matter attached to the silk but to the silk fibroin itself. PMID- 11402840 TI - Possible postcranial pneumaticity in the last common ancestor of birds and crocodilians: evidence from Erythrosuchus and other Mesozoic archosaurs. AB - Birds and crocodilians (extant archosaurs) have differing, distinctive morphologies. Birds have respiratory airsacs with diverticula that pneumatize the postcranial skeleton, a feature absent in crocodilians. Bony correlates of pneumatic sinuses are known in the vertebrae of some non-avian dinosaurs and in pterosaurs--taxa more closely related to birds than crocodilians. This and the apparent absence of pneumatic postcranial bones in fossil archosaurs more closely related to crocodilians than to birds, has been interpreted as evidence that postcranial pneumaticity is a derived character of birds and their nearest fossil relatives. The presence of apparent osteological correlates of postcranial pneumaticity is here reported in some non-crown-group archosaurs, and some of the fossil taxa more closely related to crocodilians than to birds. This suggests that the last common ancestor of birds and crocodilians might have had a pneumatized postcranium, and that the absence of this feature in crocodilians might be derived. PMID- 11402841 TI - The "Linh Duong" Pseudonovibos spiralis (Mammalia, Artiodactyla) is a new buffalo. AB - The controversial phylogenetic position of the recently described South-East Asian endemic bovid, Pseudonovibos spiralis, was evaluated on the basis of phylogenetic analyses of originally obtained nearly complete 12S mitochondrial rDNA sequences for this species and Bubalus bubalis and 26 sequences of Bovidae from the Genbank using Cervus elaphus (Cervidae) as outgroup. In most of the phylogenetic analyses performed using PAUP 4.0 (maximum parsimony, maximum likelihood and neighbour-joining), Bovidae consisted of two major clades: Bovinae including the tribes Bovini, Tragelaphini and Boselaphini, and Antilopinae + Caprinae, incorporating all other bovids. In most trees P. spiralis fell within the buffalos (subtribe Bovina) between Bubalus and Syncerus. Therefore, our phylogenetic analyses of bovid mitochondrial 12S rRNA gene sequences suggest the close relationship of this enigmatic species with the buffalos and its placement within the subtribe Bovina. PMID- 11402842 TI - Effects of bright or dim light during the daytime on digestive activity in humans. PMID- 11402843 TI - Perceptual deformation induced by visual motion. AB - The perceived position of a moving object can be misleading because the object has advanced while its previous retinal image has been transmitted through the visual stream, leading to a mismatch between actual location and its neural representation. It has been suggested that the human visual system compensates for neural processing delays to retrieve instantaneous position. However, such a mechanism would require a precise measure of the actual delay in order to provide a reliable position estimate. A novel illusory deformation of moving contours demonstrates that humans misjudge the spatial relationship between parts of coherently moving targets, and therefore do not perfectly account for neural delays. The size of this deformation increases with growing speed. In some subjects this illusion can be reversed by varying the luminance of individual dots; a manipulation that affects the neural delays. Our experiments agree with other evidence that the capacity of the visual system to compensate for processing delays is limited. PMID- 11402844 TI - Asynchronous emergence by loggerhead turtle (Caretta caretta) hatchlings. AB - For many decades it has been accepted that marine turtle hatchlings from the same nest generally emerge from the sand together. However, for loggerhead turtles (Caretta caretta) nesting on the Greek Island of Kefalonia, a more asynchronous pattern of emergence has been documented. By placing temperature loggers at the top and bottom of nests laid on Kefalonia during 1998, we examined whether this asynchronous emergence was related to the thermal conditions within nests. Pronounced thermal variation existed not only between, but also within, individual nests. These within-nest temperature differences were related to the patterns of hatchling emergence, with hatchlings from nests displaying large thermal ranges emerging over a longer time-scale than those characterised by more uniform temperatures. In many egg-laying animals, parental care of the offspring may continue while the eggs are incubating and also after they have hatched. Consequently, the importance of the nest site for determining incubation conditions may be reduced since the parents themselves may alter the local environment. By contrast, in marine turtles, parental care ceases once the eggs have been laid and the nest site covered. The positioning of the nest site, in both space and time, may therefore have profound effects for marine turtles by affecting, for example, the survival of the eggs and hatchlings as well as their sex (Janzen and Paukstis 1991). During incubation, sea turtle embryos grow from a few cells at oviposition to a self-sufficient organism at hatching some 50-80 days later (Ackerman 1997). After hatching, the young turtles dig up through the sand and emerge typically en masse at the surface 1-7 nights later, with a number of stragglers following over the next few nights (Christens 1990). This contrasts with the frequently observed pattern of hatching asynchrony in birds. It has been suggested that the cause of mass emergence in turtles is that eggs within a clutch are fertilised within a short period of time and then, when thermal conditions within the nest are uniform, develop at very similar rates and hence hatch and emerge together (Porter 1972). As a corollary of this idea, it would be predicted that when there are pronounced within-nest thermal gradients, development rates of siblings will be different and hence asynchronous hatching and emergence might occur. While it may be energetically beneficial for hatchlings to emerge in a group (Carr and Hirth 1961), if the extent of hatching asynchrony is marked then there may be severe costs for individuals if they wait for all their siblings to hatch before attempting to dig out of the sand (Hays and Speakman 1992). Under such conditions, the protracted emergence of small groups of hatchlings over several nights may be favoured. Examination of the literature suggests that emergence asynchrony may be more widespread than generally considered. For example, Witherington et al. (1990) described loggerhead turtle hatchlings (Caretta caretta) emerging over 4 days in Florida; for green turtles (Chelonia mydas), Hendrickson (1958) documented that nests in Malaysia and Sarawak produced hatchlings for up to 8 days; whilst Diamond (1976) found that hawksbill (Eretmochelys imbricata) nests on Cousin Island, Seychelles, were active for up to 4 days. Similarly, on the Greek Island of Kefalonia, we have shown that emergence from individual loggerhead turtle nests may occur on up to 11 nights (Hays and Speakman 1992). It is logical to suppose that asynchronous emergence relates to thermal gradients within nests, since the incubation duration of sea turtle eggs is related to temperature, with eggs hatching quicker when the temperature is higher. Here we test this hypothesis by measuring thermal variations within loggerhead turtle nests and comparing these variations to the patterns of hatchling emergence. PMID- 11402845 TI - Multimodular biocatalysts for natural product assembly. AB - Nonribosomal peptides and polyketides represent a large class of natural products that show an extreme structural diversity and broad pharmacological relevance. They are synthesized from simple building blocks such as amino or carboxy acids and malonate derivatives on multimodular enzymes called nonribosomal peptide synthetases (NRPSs) and polyketide synthases (PKSs), respectively. Although utilizing different substrates, NRPSs and PKSs show striking similarities in the modular architecture of their catalytic domains and product assembly-line mechanism. Among these compounds are well known antibiotics (penicillin, vancomycin and erythromycin) as well as potent immunosuppressive agents (cyclosporin, rapamycin and FK 506). This review focuses on the modular organization of NRPSs, PKSs and mixed NRPS/PKS systems and how modules and domains that build up the biosynthetic templates can be exploited for the rational design of recombinant enzymes capable of synthesizing novel compounds. PMID- 11402846 TI - [History and effect of the Kiel lymph node register. Prehistory]. PMID- 11402847 TI - [Histopathological aspects of Langerhans cell histiocytosis. With emphasis on the pulmonary form]. AB - Langerhans cell histiocytosis (LCH) occurs principally but not exclusively in younger age groups. It can present as a single lesion or as multiple lesions and can affect one or several organ systems. Diagnosis of LCH requires the identification of CD1a and S-100 positive histiocytes. The range of synonyms used for LCH reflects both the variable clinical presentation and the variations in the histological "development stage" of the individual lesions from one case to another. The term LCH has now been accepted for all manifestations of the disease. The extent of the disease in a given case is specified by determining the number and location of foci and by ascertaining whether one or several organ systems are affected, and whether organ dysfunction exists. Among cases of LCH in adults, pulmonary LCH has a special status, as this usually results from cigarette smoking and is generally limited to the lungs. The lungs may also be affected in "conventional" LCH, and organ dysfunction may also exist in this form of the disease. In general, the morphology of LCH is sufficiently characteristic that the differential diagnosis versus other types of histiocytosis should not be difficult, provided an adequate specimen is available for examination. PMID- 11402848 TI - [Histopathological diagnosis of primary liver tumors]. AB - Although modern imaging can predict the diagnosis of the vast majority of liver neoplasms, liver biopsy remains the safest, most accurate and most economical diagnostic gold standard for therapeutic implications. Refined preparation techniques using fine-needle biopsy lead to the correct diagnosis by conventional staining in most cases. Metastases and rare mesenchymal liver tumors that mimic primary hepatocellular tumors morphologically can be distinguished by use of a panel of antibodies in immunohistochemistry. Precursors of hepatocellular carcinoma (HCC) remain difficult to categorize in small biopsy specimens. Differentiation of HCC from regenerative nodules and liver cell adenomas can be achieved by molecular techniques applicable to formalin-fixed and paraffin embedded tissue (FISH or ISH). The grading and staging of resected malignant liver neoplasms according the pTNM system still have the most important prognostic impact, but screening of changes in the cell cycle by immunohistochemistry and application of molecular techniques may prove to have prognostic value in the near future. PMID- 11402849 TI - [Clinical and genetic aspects of hereditary hemochromatosis]. AB - Hereditary hemochromatosis (HH) is an autosomal recessive disease in which increased iron absorption causes iron overload and irreversible tissue damage. As laboratory parameters for measuring HH lack specificity, and HH remains asymptomatic for a long time, methods for genetic screening are highly valuable. Mutations in two genes, hfe and TfR2, have recently been found to be responsible for HH. The mutation C282Y in the hfe gene is detected in 70-95% of German and Austrian HH patients. Mutations in the TfR2 gene have been detected only very recently, and results of larger epidemiological studies are not yet available. Molecular methods permit molecular diagnosis and genetic screening. PMID- 11402850 TI - [Pulmonary lymphangioleiomyomatosis. Morphologic and immunohistochemical findings]. AB - Pulmonary lymphangioleiomyomatosis (LAM) is a rare disease affecting only women which is characterised by cystic parenchymal changes and smooth muscle proliferation and has to be considered in the differential diagnosis of interstitial lung disease. The LAM cell is classified as a so-called perivascular epithelioid cell (PEC) showing immunohistochemical co-expression of smooth muscle and melanocytic markers (melanomyopericyte). A total of 18 cases of LAM and 56 cases of various other pulmonary diseases were analysed by immunohistochemistry. For the diagnosis of LAM in transbronchial or open lung biopsies, immunohistochemistry using the antibody HMB 45 is decisive because it shows a highly specific and sensitive, but often only faint reaction for pulmonary LAM. The immunohistochemical detection of nuclear localised microphthalmia transcription factor (MiTF) in LAM as an additional melanocytic marker further emphasises the melanocytic (as well as smooth muscle) line of differentiation within the cells of LAM. Using the concept of a melanomyopericyte, pulmonary LAM could be classified as a hamartomatous interstitial lung disease showing perivascular smooth muscle and partial melanocytic differentiation. PMID- 11402851 TI - [Expression of galectin-3 in thyroid gland and follicular cell tumors of the thyroid. A critical study of its possible role in preoperative differential diagnosis]. AB - Galectin-3 belongs to a group of endogenous lectins with an affinity to glycoconjugates containing beta-galactoside residues. It has been detected in numerous tissues and studied in connection with tumor growth, dedifferentiation, and metastasis. Only few studies have dealt with galectin-3 detection in tumors of the thyroid gland and with its possible role for differential diagnosis. We studied 118 cases of thyroid gland tumors with a monoclonal antibody against galectin-3; we compared the preparations by a semiquantitative score to determine differences in expression. Normal thyroid gland tissue, goiter tissue, and tissue with functional enhancement were largely negative for galectin-3. Adenomas with a typical cytological pattern were predominantly negative, but a focal positive reaction in single cells and cell groups or follicles was possible. Almost all papillary carcinomas showed a distinct galectin-3 expression. While findings in follicular carcinomas and oxyphilic adenomas and carcinomas were very uneven, with both positive and negative tumors, the galectin-3 reaction can be helpful in recognizing follicular variants of the papillary carcinoma. Investigation of the biological significance of tumors should always be cautious and consider known histological criteria for malignancy. PMID- 11402852 TI - [Desmosis coli in adults. Differential diagnosis of chronic constipation]. AB - Desmosis coli is a disturbance of the intramural connective tissue mesh network of the colonic wall which can lead to a hypoperistalsis syndrome with chronic constipation in the absence of any anomaly of the vegetative gut innervation. The condition typically occurs in infants and adolescents; however, as an incomplete form, desmosis coli can also cause chronic constipation in adults, as demonstrated in this case report. PMID- 11402853 TI - [Structural changes in pathology. The Institute of Pathology in Essen-Steele]. PMID- 11402854 TI - Comparative pathogenesis of acute and latent infections of calves with bovine herpesvirus types 1 and 5. AB - This study was conducted to compare the pathogenesis of acute and latent infections with closely related bovine herpesvirus types 1 (BHV-1) and 5 (BHV-5) in their natural host. Two groups of eight calves were inoculated intranasally with BHV-1 or BHV-5. Although BHV-1 and BHV-5 similarly replicate in the nasal mucosa after inoculation, both viruses differ markedly in their ability to cause disease, BHV-5 being responsible of some fatal encephalitis while BHV-1 inducing rhinotracheitis. Virus isolation and immunohistochemistry demonstrated that BHV-5 replicates extensively in neurons of the central nervous system (CNS) and in respiratory cells of lungs, tracheal and nasal mucosae. Invasion of the CNS likely occurs through the trigeminal and olfactory pathways. Both groups developed cross-neutralising antibodies during this experiment suggesting partial clinical cross-protection afforded by the two infections. Three months after primary infection, experimental reactivation showed that BHV-5 was able to establish latency in the trigeminal ganglia but also the CNS of surviving calves. Moreover, laboratory findings suggested that BHV-5 could also persist in the tracheal and nasal mucosae. These results indicate that, after primary infection, BHV-1 and BHV-5 displayed similar biological features and consequently need to be considered together for the control of BHV-1 infection. PMID- 11402855 TI - The formation of empty shells upon pressure induced decapsidation of turnip yellow mosaic virus. AB - The stability of turnip yellow mosaic virus (TYMV) was investigated under pressure, using solution neutron small angle scattering. Dissociation products were characterized by analytical ultracentrifugation and electron microscopy. At pH 6.0, TYMV remained unaffected by pressure, up to 260 Megapascals (MPa), the highest pressure reached in these experiments. At pH 8.0, TYMV remained unaffected by pressure up to 160 MPa, but decapsidated irreversibly above 200 MPa, giving rise to more and more empty shells upon increasing pressure. The organization of these empty shells was similar to that of the capsid of native virions, apart from the presence of a hole corresponding to the loss of a group of 5-8 coat protein subunits, through which the RNA may have escaped. At variance with other small isometric viruses, the capsid of TYMV never dissociated under pressure into subunits or small aggregates of subunits. This exceptional behavior of TYMV is probably due to the importance of van der Waals contacts and hydrogen bonds in the stability of its capsid. PMID- 11402856 TI - Reduced expression of surface glycoproteins in mouse fibroblasts persistently infected with human respiratory syncytial virus (HRSV). AB - BCH4 cells, persistently infected with Human Respiratory Syncytial Virus (HRSV), were obtained by Fernie et al. [12] after infection of a BALB/c mouse embryo cell line with the Long strain of HRSV. To understand the basis of HRSV persistence, the expression of HRSV RNAs and proteins was evaluated in BCH4 cells and infected parental BALB/c and fully permissive HEp-2 cells. Production of viral mRNAs was severely impaired in BCH4 cells. In addition, the expression level of the surface glycoproteins F and G was markedly reduced relative to internal viral proteins. However, virus recovered from BCH4 cells could lytically infect HEp-2 cells and expressed normal levels of surface glycoproteins. No evidence of defective genomes or interfering particles was found in BCH4 cells. Taken together, these data indicate that reduction of both viral mRNA accumulation and surface glycoprotein biosynthesis are at the basis of HRSV persistence in BCH4 cells. PMID- 11402857 TI - Combination of amino acids in the 3a protein and the coat protein of cucumber mosaic virus determines symptom expression and viral spread in bottle gourd. AB - Bottle gourd plants infected with an isolate of cucumber mosaic virus (CMV-KM) developed severe chronic mosaic symptoms (SCMS) with stunting, but two other isolates (CMV-Y and CMV-D8) did not. CMV-KM and CMV-D8 induced enlarged chlorotic spots and rapidly spread over the inoculated cotyledons, whereas CMV-Y elicited a hypersensitive response (HR) producing pin-point necrotic lesions. Reassortment analysis among the three isolates revealed that the local and systemic symptoms on the plants were regulated by RNA3. Reciprocal recombination and site-directed point mutation analyses of the three RNA3s demonstrated that a combination of genetic information encoded by the movement protein (MP) gene and the coat protein (CP) gene determines the induction of SCMS in bottle gourd. SCMS occurred when Ser51 in the MP of CMV-D8 was changed to Asn51, whereas substitution of Ser51 for Asn51 in the MP of CMV-KM eliminated its ability to induce SCMS. Furthermore, Ser129 in the CPs was shown to be responsible for induction of HR and blocking of efficient cell-to-cell and long-distance movement. PMID- 11402858 TI - Characterisation of a chicken anaemia virus variant population that resists neutralisation with a group-specific monoclonal antibody. AB - A variant population of chicken anaemia virus (CAV), termed P310 2A9-resist, that resists neutralisation by the monoclonal antibody (MAb) 2A9, was selected from Cux-1 virus that had been passaged 310 times (P310) in MDCC-MSB1 cells. Substantially higher concentrations of MAb 2A9 were required to neutralise the selected virus compared to those required to neutralise a low-passage (P13) Cux-1 isolate. Virus neutralisation tests showed that serum from chickens infected with the P310 2A9-resist virus neutralised P13 virus and that serum from chickens infected with P13 virus conversely neutralised the P310 2A9-resist virus. MDCC MSB1 cells infected with the P310 2A9-resist virus produced no staining with low dilutions (1:100) of Mab 2A9 in an indirect immunofluorescence (IF) test, whereas cells infected with P13 virus reacted positively at high MAb dilutions (1:80,000). Experimental infections of 1-day-old SPF chicks showed that the P310 2A9-resist virus was substantially attenuated. Chimaeric viruses constructed using PCR-amplified regions from the P310 2A9-resist and a pathogenic low-passage cloned Cux-1 isolate showed that the reduced MAb reactivity and attenuation exhibited by the P310 2A9-resist virus were mainly associated with a region encoding the N-terminal half of the 50 kDa capsid protein VP1 and C-terminal regions of VP2 and VP3. The nucleotide sequence of the protein-coding region of the P310 2A9-resist virus is reported and the amino acid sequences of the 3 encoded proteins compared with those of other Cux-1 isolates. PMID- 11402859 TI - Down-regulation of translation driven by hepatitis C virus internal ribosomal entry site by the 3' untranslated region of RNA. AB - The genome of hepatitis C virus (HCV) is a single-stranded RNA of positive polarity that has a poly(U/C) tract followed by a highly conserved 98-nt sequence, termed the X region, in the 3' untranslated region (UTR). To investigate the effect of the 3'UTR on the HCV translation that depends on the internal ribosomal entry site (IRES), we prepared a deletion HCV RNA, MA delta, that lacked the RNA region from nt 1286 to 8785. A series of MA delta RNAs that differ in the primary structure of their 3'UTR, were generated and examined for their translation efficiencies in reticulocyte lysates. Deletion of the poly(U/C) tract and/or stem-loop structure (SL) 3 region of 3'X resulted in enhancement of the translation efficiency. Translation of MA delta RNA was inhibited by the addition of recombinant polypyrimidine tract-binding protein (PTB). A similar inhibition by PTB, however, was observed when an RNA lacking the poly(U/C) tract or SL3 region was used. The inhibitory effect by PTB was not obvious for MA delta (1041) RNA composed of nt 1 to 1041 but MA delta (8928) RNA composed of nt 1 to 1285 and nt 8786 to 8928. These results suggest that the observed down-regulation of HCV translation by the 3'UTR is mediated by some host factor(s) other than PTB, and that a PTB site for inhibition resides in the coding sequence of nt 1042 to 8928 of MA delta RNA. PMID- 11402860 TI - A single amino acid change at Leu-188 in the reverse transcriptase of HIV-2 and SIV renders them sensitive to non-nucleoside reverse transcriptase inhibitors. AB - Non-nucleoside reverse transcriptase inhibitors (NNRTIs) are selective for human immunodeficiency virus type 1 (HIV-1) and generally not effective on HIV-2 or simian immunodeficiency virus (SIV). Only SIVagm was found to be sensitive to NNRTIs. When the amino acid differences in RT between SIVmac and SIVagm were compared with the known amino acid substitutions of NNRTI-resistance variants of HIV-1, we came to consider that the amino acid residue Leu-188 of HIV-2 and SIVmac might be related to their resistance to NNRTIs. To test this hypothesis, we substituted Leu-188 to Cys or Tyr in HIV-2 and SIVmac, and examined sensitivity of the mutant molecular clones to NNRTIs. The L188Y mutant of HIV-2 became completely sensitive to delavirdine and efavirenz, while that of SIVmac was also significantly sensitive to these NNRTIs. We further isolated NNRTI resistant variants from these mutant viruses and determined amino acid substitutions in RT. The roles of the observed substitutions in NNRTI-resistance were further confirmed by site-directed mutagenesis. Our study reveals the crucial role of L188 in the natural resistance of HIV-2 and SIVmac to NNRTIs. Furthermore, the observed substitutions in RT of HIV-2 and SIVmac support the common mechanism of action of NNRTIs against HIV-1, HIV-2 and SIV. PMID- 11402861 TI - A universal PCR primer to detect members of the Potyviridae and its use to examine the taxonomic status of several members of the family. AB - A universal primer (Sprimer: 5'-GGX AAY AAY AGY GGX CAZ CC-3', X = A, G, C or T; Y = T or C; Z = A or G), designed from the consensus sequences that code for the conserved sequence GNNSGQP in the NIb region of members of the family Potyviridae, was used to amplify by RT-PCR the 3'-terminal genome regions from infected plant samples representing 21 different viruses in the family. Sequencing of some of the fragments (c. 1.7 kb) showed that the type strain (ATTC PV-107) of Oat necrotic mottle virus is not a distinct species in the genus Rymovirus, but is synonymous with Brome streak mosaic virus (genus Tritimovirus) and that Celery mosaic virus is a distinct member of the genus Potyvirus not closely related to any other sequenced species. Potyviruses infecting crops in China were also investigated, showing that viruses on cowpea and maize in Hangzhou, Zhejiang province were respectively Bean common mosaic virus and Sugarcane mosaic virus and that one on garlic in Nanjing, Jiangsu province was Onion yellow dwarf virus. Fragments were also sequenced from Chinese isolates of Lettuce mosaic virus and Soybean mosaic virus (from Hangzhou), Turnip mosaic virus (2 different isolates from Zhejiang province) and RNA1 of Wheat yellow mosaic virus (from Rongcheng, Shandong province). PMID- 11402862 TI - Replication of Chilo iridescent virus in the cotton boll weevil, Anthonomus grandis, and development of an infectivity assay. AB - The boll weevil, Anthonomus grandis Boheman, is a devastating pest of cotton. Chemical pesticides are problematic due to relative lack of target specificity and resistance. Microbial pesticides may provide viable alternatives because of their narrow host range. Chilo iridescent virus (CIV) is the type species for genus Iridovirus, family Iridoviridae: large, icosahedral cytoplasmic viruses containing a double-stranded DNA genome. Earlier work suggested that CIV replicated in the boll weevil; however, efficiency or production of infectious virus was not established. We showed that CIV undergoes a productive cycle in A. grandis. CIV DNA levels in boll weevil pupae increased significantly from 0 to 3 days post infection. Moreover, virogenic stromata and complete virus particles were observed in the cytoplasm by 7 days. An endpoint dilution assay using viral DNA replication as indicator suggested a 10(5)-fold increase in infectious virus titer over 7 days. This is the first such demonstration in larval infections with genus Iridovirus. Our study establishes that CIV undergoes a productive cycle in the boll weevil and provides an important and useful model system for replication at the organismal level. These results have important implications for the potential of CIV and its components in boll weevil control. PMID- 11402863 TI - Effect of fatty acids on arenavirus replication: inhibition of virus production by lauric acid. AB - To study the functional involvement of cellular membrane properties on arenavirus infection, saturated fatty acids of variable chain length (C10-C18) were evaluated for their inhibitory activity against the multiplication of Junin virus (JUNV). The most active inhibitor was lauric acid (C12), which reduced virus yields of several attenuated and pathogenic strains of JUNV in a dose dependent manner, without affecting cell viability. Fatty acids with shorter or longer chain length had a reduced or negligible anti-JUNV activity. Lauric acid did not inactivate virion infectivity neither interacted with the cell to induce a state refractory to virus infection. From mechanistic studies, it can be concluded that lauric acid inhibited a late maturation stage in the replicative cycle of JUNV. Viral protein synthesis was not affected by the compound, but the expression of glycoproteins in the plasma membrane was diminished. A direct correlation between the inhibition of JUNV production and the stimulation of triacylglycerol cell content was demonstrated, and both lauric-acid induced effects were dependent on the continued presence of the fatty acid. Thus, the decreased insertion of viral glycoproteins into the plasma membrane, apparently due to the increased incorporation of triacylglycerols, seems to cause an inhibition of JUNV maturation and release. PMID- 11402864 TI - Feline and canine coronaviruses are released from the basolateral side of polarized epithelial LLC-PK1 cells expressing the recombinant feline aminopeptidase-N cDNA. AB - In this study feline (FECV and FIPV) and canine (CCoV) coronavirus entry into and release from polarized porcine epithelial LLC-PK1 cells, stably expressing the recombinant feline aminopeptidase-N cDNA, were investigated. Virus entry appeared to occur preferentially through the apical membrane, similar to the entry of the related porcine coronavirus transmissible gastroenteritis virus (TGEV) into these cells. However, whereas TGEV is released apically, feline and canine coronaviruses were found to be released from the basolateral side of the epithelial cells. These observations indicate that local infections as caused by TGEV, FECV and CCoV do not strictly correlate with apical release, as suggested by earlier work. PMID- 11402865 TI - Group C rotavirus NSP4 induces diarrhea in neonatal mice. AB - Nonstructural glycoprotein NSP4 of group A rotavirus induces diarrhea in neonatal mice by functioning as an enterotoxin. Previously, our laboratory reported that the structural features of group A and group C rotavirus NSP4 proteins are well conserved despite a lack of sequence homology between group A and group C rotavirus NSP4 proteins [Horie Y, et al., Arch Virol (1997) 142: 1865-1872]. To test whether group C rotavirus NSP4 has an enterotoxigenic activity, we expressed in Escherichia coli the carboxy two-thirds (corresponding to amino acid residues 55-150) of the NSP4 protein derived from group C rotavirus strain Ehime 9301. This truncated NSP4 protein was able to induce diarrhea in 5-day-old CD-1 mice when administered intraperitoneally. Thus, group C rotavirus NSP4 acts as an enterotoxin like group A rotavirus NSP4. PMID- 11402866 TI - Sequence diversity and interrelationships among isolates of satsuma dwarf-related viruses. AB - The 3'-region of RNA2 of three viruses (Natsudaidai dwarf virus (isolate ND-1), and two unidentified isolates (LB-1, Az-1)), which were related to Satsuma dwarf virus (SDV), were sequenced. Phylogenetic analysis including the previously reported SDV-related viruses (Citrus mosaic virus (CiMV, Ci-968), Navel orange infectious mottling virus (NIMV, NI-1)) showed that they were classified into three groups, SDV (S-58), CiMV (Ci-968, LB-1, Az-1, ND-1), and NIMV (NI-1). The results suggested these groups might correspond to the three distinct virus species. ND-1, LB-1, and Az-1 were considered strains of CiMV, although they do not induce citrus mosaic on the fruit rind. PMID- 11402867 TI - Measles virus circulation in Argentina: 1991-1999. AB - Nucleoprotein (N) and Haemagglutinin (H) genes from measles viruses isolated from Argentina before and after the 1993 and 1998 massive vaccination campaigns were characterised to determine genetic variations that occurred from 1991 to 1999. Measles viruses from the 1991-94 period were clustered with the C1 genotype and those from 1997-99 with D6. Genetic variations within the 1997-99 outbreak were less than 1.2% and 0.79% for the N and H sequences respectively. The C1 genotype has not been detected since 1994 and the finding that a single D6 virus was found in November 1999 demonstrates that wild type viruses are still circulating among a partially covered population. PMID- 11402868 TI - The RNA 5 of Prunus necrotic ringspot virus is a biologically inactive copy of the 3'-UTR of the genomic RNA 3. AB - In addition to the four RNAs known to be encapsidated by Prunus necrotic ringspot virus (PNRSV) and Apple mosaic virus (ApMV), an additional small RNA (RNA 5) was present in purified preparations of several isolates of both viruses. RNA 5 was always produced following infection of a susceptible host by an artificial mixture of RNAs 1, 2, 3, and 4 indicating that it was a product of viral replication. RNA 5 does not activate the infectivity of mixtures that contain the three genomic RNAs (RNA 1 + RNA 2 + RNA 3) nor does it appear to modify symptom expression. Results from hybridization studies suggested that RNA 5 had partial sequence homology with RNAs 1, 2, 3, and 4. Cloning and sequencing the RNA 5 of isolate CH 57/1-M of PNRSV, and the 3' termini of the RNA 1, RNA 2 and RNA 3 of this isolate indicated that it was a copy of the 3' untranslated terminal region (3'-UTR) of the genomic RNA 3. PMID- 11402869 TI - Isolation and characterization of porcine circovirus 2 from cases of sow abortion and porcine dermatitis and nephropathy syndrome. AB - We report the isolation and characterisation of porcine circovirus 2 (PCV2) from cases of sow abortion and porcine dermatitis and nephropathy syndrome. The results suggest that the clinical scope of PCV2 infections requires continuous re evaluation. PMID- 11402870 TI - [MRI diagnosis of injuries and diseases of peroneal tendons]. AB - Injuries of the peroneus tendons are common and both the athlete and the older population are at risk. MR imaging is a useful technique for revealing injuries of the peroneus tendons as well as showing anatomic factors associated with these lesions. This article reviews clinical factors and MR imaging characteristics of injuries of the peroneus tendons. PMID- 11402871 TI - [Gorham syndrome--massive osteolysis of the cervical vertebrae]. PMID- 11402872 TI - [Detection of liver lesions: magnetic resonance tomography using superparamagnetic iron particles (AMI-25, endorem) as a contrast medium in comparison with ultrasound diagnosis, computerized tomography and CT portography]. PMID- 11402873 TI - [Pulmonary lymphangioleiomyomatosis--review and personal results]. AB - Pulmonary lymphangioleiomyomatosis (LAM) is characterised by the proliferation of smooth muscle cells in the peribronchial, perivascular and perilymphatic tissue resulting in severe destructive changes of the lung parenchyma. We reviewed the literature and compared the findings in chest radiograph and CT with our own results obtained in a group of 12 patients suffering from histological proven LAM. The main findings of LAM in chest radiograph are a diffuse reticular opacity and an increased lung volume, often large cysts are visible. The CT scans reveal that cystic lesions are present in all patients, but the reported size of the cystic lesions is contradictory. We postulate two main types of LAM: the one resulting in a diffuse distribution of multiple small cysts, partially demonstrating a honey-combing pattern, the other showing severe destruction of the lung parenchyma due to large cysts. The presence of fibrotic changes and architectural distraction does not exclude LAM. So chest radiograph as well as computed tomography reveal neither uniform nor pathognomonic findings. Differential diagnosis of LAM seems to be more difficult than most authors believe. PMID- 11402874 TI - ["Phtisis atra"--a now seldom disease picture as a special disease course of silicosis]. AB - Progressive massive fibrosis (PMF) of the lung is caused by coalescence of fibrotic nodules. The center of the PMF often displays necrotic areas. If the necrosis gets in contact to the bronchial system cavitation may occur. We report on a 68 year old patient suffering from severe silicosis of the lung and metastatic spread of a histologically proven lung cancer into the brain. The patient who was administered to the hospital under the intention of cerebral radiotherapy showed a colliquative PMF in the right upper lobe with cavitation and expectoration of large amounts of black-stained sputum (melanoptysis). PMID- 11402875 TI - [Time needs in evaluating digital thoracic images on the monitor in comparison with alternator]. AB - PURPOSE: Evaluation of time efficiency in softcopy reading versus hardcopy reading of digital chest x-rays. MATERIAL AND METHODS: 130 normal and pathologic chest x-rays in two plains were analyzed by 4 experienced radiologist at both a digital workstation and the light box. Reading time and switch time between two patient folders were measured as well as the frequency of post-processing at the monitor. RESULTS: Reading time at the workstation slightly exceeded reading time at the light box; differences were not statistically significant. Post-processing (frequency between 2% and 83%) did not significantly prolong reading time. Mean switch time between two patient folders was 4.3 sec at the workstation and 13.7 sec at the light box. CONCLUSION: As compared to hardcopy reading, softcopy reading of digital chest x-rays does not significantly increase reading time. Switch time between patient folders can be reduced at the workstation by a factor of two to four. PMID- 11402876 TI - An update on functional electrical stimulation after spinal cord injury. AB - Recent advances in biomedical engineering as applied to neurologic rehabilitation have finally borne clinically relevant fruit. Nowhere is this more evident than in the field of functional electrical stimulation (FES). This article highlights the remarkable clinical progress that has been made in the use of electrical stimulation for restoring movement and function in individuals with spinal cord injury. Specific attention is given to respiratory-assist devices, hand-grasp systems, standing and walking, and bladder control. This review article features discussion of eight devices that have gone through the United States Food and Drug Administration (FDA) regulatory approval process. PMID- 11402877 TI - Neurotrophic factors and gene therapy in spinal cord injury. AB - Although it was once thought that the central nervous system (CNS) of mammals was incapable of substantial recovery from injury, it is now clear that the adult CNS remains responsive to various substances that can promote cell survival and stimulate axonal growth. Among these substances are growth factors, including the neurotrophins and cytokines, and growth-supportive cells such as Schwann cells, olfactory ensheathing glia, and stem cells. We review the effects of these substances on promoting axonal growth after spinal cord injury, placing particular emphasis on the genetic delivery of nervous system growth factors to specific sites of injury as a means of promoting axonal growth and, in limited instances, functional recovery. PMID- 11402878 TI - Phantom finger phenomena and the effects of toe-to-finger transplantation. AB - Phantom finger phenomena and the effects of toe-to-finger transplantation were studied in 76 patients who had had traumatic finger amputation. Phantom finger phenomena were observed in 48 (63%) patients with the presence of phantom finger only in 30, phantom finger with sensation in nine, and phantom finger with motion also in nine. After toe transplantation, phantom finger phenomena disappeared immediately in about half of the transplanted fingers that had phantom phenomena before toe transplantation, and also in about half of the amputated fingers without the surgery. Conversely, phantom toe phenomena occurred in 13 (17%) patients. Although some patients had mild-to-moderate unpleasant phantom sensations, none had severe or distressing phantom finger pain or phantom toe pain. It is concluded that phantom phenomena occurred in both finger and toe amputations, and that toe-to-finger transplantation appeared to facilitate the disappearance of phantom phenomena not only in the transplanted fingers but also in the amputated but untransplanted fingers. Possible mechanisms for these observations are discussed. PMID- 11402879 TI - An analysis of changes in sensory thresholds to mild tactile and cold stimuli after experimental spinal cord injury in the rat. AB - Changes in sensory function including chronic pain and allodynia are common sequelae of spinal cord injury (SCI) in humans. The present study documents the extent and time course of mechanical allodynia and cold hyperalgesia after contusion SCI in the rat using stimulation with graded von Frey filaments (4.97 50.45 g force) and ice probes. Fore- and hind-paw withdrawal thresholds to plantar skin stimulation were determined in rats with a range of SCI severities (10-g weight dropped from 6.25, 12.5, or 25 mm using the MASCIS injury device); animals with 25-mm injuries most consistently showed decreased hind-paw withdrawal thresholds to touch and cold, which developed over several weeks after surgery. Stimulation of the torso with graded von Frey hairs was performed at specified locations on the back and sides from the neck to the haunch. Suprasegmental responses (orientation, vocalization, or escape) to mechanical stimulation of these sites were elicited infrequently in the laminectomy control rats and only during the first 3 weeks after surgery, whereas in 25-mm SCI rats, such responses were obtained for the entire 10 weeks of the study. These data suggest that rats with contusion SCI may exhibit sensory alterations relevant to human spinal cord injuries. PMID- 11402880 TI - Factors contributing to motor impairment and recovery after stroke. AB - The goal of the present study was to examine factors affecting motor impairment and recovery in a primate model of cortical infarction. Microelectrode stimulation techniques were used to delineate the hand representation in the primary motor cortex (M1). Microinfarcts affecting approximately 30% of the hand representation were made by electrocoagulation of surface vessels. Electrophysiologic procedures were repeated at 1 month after the infarct to examine changes in motor map topography. Before the infarct, and at approximately 1 week (early period) and 1 month (late period) after the infarct, manual performance was assessed on a reach-and-retrieval task that required skilled use of the digits. Contrary to the expected outcome, early impairment was inversely related to the amount of digit representation destroyed by the infarct. That is, animals with less involvement of the M1 digit area demonstrated the greatest motor deficit in the early postinfarct period. In addition, improvement in motor performance between early and late postinfarct periods was directly related to a decrease in the extent of the digit + wrist/forearm area in the final postinfarct map. These results suggest that specific aspects of motor-map remodeling are expressions of adaptive mechanisms that underlie functional recovery after stroke. Further, they suggest that the adaptive mechanisms underlying postinjury recovery differ in detail from those that operate in normal motor learning. The potential role of compensatory mechanisms in these phenomena is discussed. PMID- 11402881 TI - Fibroblasts genetically modified to produce BDNF support regrowth of chronically injured serotonergic axons. AB - Cells genetically modified to release a variety of growth and/or neurotrophic factors have been used for transplantation into the injured spinal cord as a means to deliver therapeutic products. Axon growth into and through such transplants has been demonstrated after intervention after an acute injury. The present study examined their potential to support regeneration in a chronic injury condition. Five weeks after a cervical hemisection in adult rats, the lesion site was debrided of scar tissue and expanded in both rostral and caudal directions. Animals received a transplant of cultured normal fibroblasts (control) or fibroblasts genetically modified to produce brain-derived neurotrophic factor (BDNF). Six weeks later, animals were killed to determine the extent of growth of serotonergic axons into the transplant. Axons immunoreactive for serotonin (5-HT-ir) were found to cross the rostral interface of host spinal cord readily with either type of fibroblast cell transplant, but the number and density of 5-HT-ir axons extending into the BDNF-producing transplants was markedly greater than those in the control fibroblasts. Axons coursed in all directions among normal fibroblast transplants, whereas growth was more oriented along a longitudinal plane when BDNF was being released by the transplanted cells. The length of growth and the percentage of the transplant length occupied by 5-HT-ir axons were significantly greater in BDNF-producing transplants than in the normal fibroblasts. Many serotonergic axons approached the caudal end of the BDNF-producing cell transplants, although most failed to penetrate the host spinal cord distal to the lesion. These results indicate that whereas fibroblast cell transplants alone can support regrowth of axons from chronically injured supraspinal neurons, modification of these cells to produce BDNF results in a significant increase in the extent of growth into the transplant. PMID- 11402882 TI - Single injections of a DNA plasmid that contains the human Bcl-2 gene prevent loss and atrophy of distinct neuronal populations after spinal cord injury in adult rats. AB - Spinal cord injury in adult mammals causes atrophy or loss of axotomized neurons. We have previously found that the product of the antiapoptotic gene Bcl-2, delivered by intraspinal injection of a DNA plasmid, reduces atrophy and loss of axotomized Clarke's nucleus neurons in adult rats. Here we studied whether the same treatment protects axotomized red nucleus (RN) neurons. Two months after the right dorsolateral funiculus was ablated in adult Sprague-Dawley rats by C3/C4 subtotal hemisection, there was approximately 48% loss of RN neurons in the magnocellular portion of the RN contralateral to the lesion and atrophy of many surviving neurons. When a DNA plasmid encoding the human Bcl-2 gene and the bacterial reporter gene LacZ, complexed with cationic lipids, was injected just rostral to the subtotal hemisection site, 87% of RN neurons survived, and there was partial, but robust, protection from atrophy. These and our previous results indicated that intraspinal administration of the Bcl-2 gene can prevent retrograde cell loss and reduce atrophy of axotomized RN and Clarke's nucleus neurons in adult rats and provide an effective means to rescue neurons whose survival depends on different growth factors. PMID- 11402883 TI - Effects of sacrocaudal spinal cord transection and transplantation of fetal spinal tissue on withdrawal reflexes of the tail. AB - Reflex responses to electrocutaneous stimulation of the tail were characterized in awake cats, before and after transection of the spinal cord at sacrocaudal levels S3-Ca1. Consistent with effects of spinal transection at higher levels, postoperative cutaneous reflexes were initially depressed, and the tail was flaccid. Recovery ensued over the course of 70-90 days after sacrocaudal transection. Preoperative and chronic postlesion reflexes elicited by electrocutaneous stimulation were graded in amplitude as a function of stimulus intensity. Chronic postlesion testing of electrocutaneous reflexes revealed greater than normal peak amplitudes, peak latencies, total amplitudes (power), and durations, particularly for higher stimulus intensities. Thus, sacrocaudal transection produced effects representative of the spastic syndrome. In contrast, exaggerated reflex responsivity did not develop for a group of cats that received transplants of fetal spinal cord tissue within sacrocaudal transection cavities at the time of injury, in conjunction with long-term immunosuppression by cyclosporine. We conclude that gray matter replacement and potential neuroprotective actions of the grafts and/or immunosuppression prevent development of the spastic syndrome. This argues that the spastic syndrome does not result entirely from interruption of long spinal pathways. PMID- 11402884 TI - Activation of supplementary motor area during imaginary movement of phantom toes. AB - To evaluate changes in the human cerebral cortex after lower limb amputation, we studied repetitive toe movements using functional magnetic resonance imaging. The subject did not experience any phantom pain but had a vivid sensation of the phantom limb's presence and was able to imagine the movement of her phantom toes and ankle. Actual movement of her normal limb activated the contralateral supplementary motor area (SMA), the primary motor cortex (M1), and the primary somatosensory cortex (S1). Movement of her phantom limb activated the contralateral SMA and the M1. Imaginary movement of her normal toes without actual movement activated the contralateral SMA. The slice level that was activated by the movement of the phantom limb was shifted 8 mm caudally, suggesting that cortical reorganization had occurred after the lower limb amputation. PMID- 11402885 TI - Should we operate on all patients with advanced gastric cancer? PMID- 11402886 TI - The attitudes of parents to traditional medicine and the surgeon. PMID- 11402887 TI - Celecoxib--the debate rages on. PMID- 11402888 TI - Appalling ultrasound knowledge? PMID- 11402889 TI - Stroke in the young in South Africa. PMID- 11402890 TI - Vaginal foreign body. PMID- 11402891 TI - Sharp eyes on cholera flashpoints. PMID- 11402892 TI - S.A. team to train Indian rescuers. PMID- 11402893 TI - Judasa's allocation alternative for C.S. PMID- 11402894 TI - HPCSA AIDS cases under scrutiny. PMID- 11402895 TI - Understaffing at rural hospitals. PMID- 11402896 TI - 1st Congress of the SA Heart Association. Stellenbosch, 26-29 November 2000. PMID- 11402897 TI - So you want to surf the Internet? Birth and rise of the Web. Part 1. PMID- 11402898 TI - ICU care in AIDS patients--is there any benefit? PMID- 11402899 TI - Hypercalcaemia and bony lesions in association with parathyroid and prostatic carcinoma. PMID- 11402900 TI - Presence of malice--Hanson redivivus. PMID- 11402901 TI - Penetrating nail gun injury of the head and chest with incidental pericallosal artery aneurysm. PMID- 11402902 TI - Nosocomial endocarditis due to extended-spectrum beta-lactamase-producing Klebsiella pneumoniae in a child. PMID- 11402903 TI - Genetic factors in pterygium in South Africans. PMID- 11402904 TI - A comparison of the perinatal mortality rates for infants weighing 500 g or more at birth and those weighing 1,000 g or more. PMID- 11402905 TI - Implications of the thrifty phenotype hypothesis for the health of societies undergoing acculturation--lessons for South African health planning. PMID- 11402906 TI - Compulsory community service for doctors in South Africa--an evaluation of the first year. PMID- 11402907 TI - Incidence and nature of epidemiological injuries to elite South African cricket players. AB - OBJECTIVES: The study investigated the incidence and nature of injuries sustained by elite cricketers during a season and the possible risk factors associated with these injuries. METHODS: Fourteen physiotherapists and three doctors recorded 163 injuries sustained by 88 cricketers. RESULTS: Bowling accounted for 40.5% of the injuries, while fielding and batting accounted for 25.6% and 21.5% of the injuries, respectively. The injuries occurred while practising for or playing in club (3.7%), provincial 'B' (8.0%), provincial (51.5%) and international (36.8%) matches and occurred primarily during 4- and 5-day (33.1%) and 1-day (27.0%) matches. First-time injuries made up 63.6% of the injuries, recurrent injuries from previous seasons made up 28.4% and 8.0% of the injuries recurred again at a later stage during the season. Of the injuries 62.6% were acute, 14.7% chronic and 22.7% were acute injuries on existing chronic injuries. The majority of the injuries were muscle (37.4%), joint (20.9%) and tendon (15.3%) injuries. The primary mechanisms of injury were as a result of overuse (30.7%); running to catch or field (15.3%); during the bowling delivery or follow-through (12.3%); and being struck by the ball while batting (11.0%). CONCLUSIONS: Coaches, players and the medical support team need to be aware of the incidence, nature and cause of injuries, so that the risk of injury can be further reduced. PMID- 11402908 TI - Cost-effectiveness of hepatitis B vaccination in haemodialysis patients. AB - BACKGROUND: Vaccination against hepatitis B virus is an important means of controlling the infection, but its role in haemodialysis patients has been questioned due to the latter's impaired immune response. METHODS: Forty-eight of 79 haemodialysis patients who were negative for antibodies to both hepatitis B surface and core antigens were entered into a vaccination programme. Standard doses of a plasma-derived vaccine were administered into the deltoid muscle at 0, 1, 2 and 4 months, and the antibody response was measured at 1 and 2 months after the third and fourth doses. RESULTS: The peak mean antibody titre of 372 IU/l was recorded at 1 month after the fourth dose, and the maximum response rate was achieved at 2 months after the final dose. Seroconversion occurred in 26 of 36 patients (72%) who completed the programme, and protective levels of antibody above 10 IU/l were found in 25 of 36 patients (69%). Cost analysis of the project revealed a net saving of +/- R90/patient entered at the end of the first year, due to the reduced number of patients requiring monthly surveillance tests for hepatitis B surface antigen. After that, an annual saving of +/- R380/patient is projected. CONCLUSION: In view of the high prevalence of chronic hepatitis B carriers in the South African population, the reduction in the number of patients at risk of infection, combined with a net cost saving, makes it reasonable to recommend vaccination in all non-immune haemodialysis patients despite a reduced response rate. PMID- 11402909 TI - Tumours and cancers in Graeco-Roman times. AB - In Graeco-Roman times all tumours (Greek: onkoi, abnormal swellings) were considered to be of inflammatory origin, the result of unfavourable humoural fluxes, and caused by an extravascular outpouring of fluid into tissue spaces. The neoplastic nature of tumours is a more recent concept, barely two centuries old. In Hippocratic literature tumours were mainly classified as karkinomata, phumata, and oidemata. Phumata included a large variety of tumours, inflammatory and neoplastic in origin, and mostly benign (in modern terms), while oidemata were soft, painless tumours and even included generalised oedema (dropsy). Although all categories possibly included occasional cancers, the vast majority of what appears to have been malignant tumours were called karkinoi karkinomata (Latin: cancrum/carcinoma). There was, however, no recognition of benign and malignant, primary and secondary tumours, in the modern sense. PMID- 11402910 TI - [Change from ACE inhibitor, Ca-antagonist or beta-blocker to candesartan cilexetil: better efficacy and tolerance. SWITCH study (German study segment)]. AB - BACKGROUND AND OBJECTIVE: Lack of efficacy in the treatment of hypertension with only one drug presents a problem in general practice and often requires switching to another type of drug, because higher dosage of currently used antihypertensives increases the frequency of side effects. Angiotensin II antagonists are well tolerated and there is no evidence of dose-related increase in side effects. This study in 574 hypertensives under therapy with ACE inhibitors, beta-blockers or calcium channel blockers was undertaken to determine whether direct switching to the Angiotensin II-antagonist candesartan cilexetil at its maximal dose of 16 mg is as effective and tolerable as starting therapy with candesartan cilexetil 8 mg followed by up-titration to 16 mg after 4 weeks. PATIENTS AND METHODS: 258 men (mean age 57 +/- 11 years) and 316 women (58 +/- 12) with essential hypertension (blood pressure < 180/95 mm Hg) under ambulatory therapy with ACE-inhibitors, beta-blockers or calcium channel blockers with inadequate efficacy or tolerability were switched to monotherapy with candesartan cilexetil. Half of the patients were treated with 8 mg for 4 weeks (n = 284), the other half received 16 mg (n = 290). Both groups then were treated with candesartan cilexetil, 16 mg, for further 4 weeks. Choice of treatment was doubly blinded and randomised. RESULTS: After 4 weeks significant blood pressure reduction was observed in both treatment groups (p < 0.0001 for each pretreatment group). A tendency for more adequate blood pressure reduction under initial therapy with candesartan cilexetil 16 mg was observed. There was a small further blood pressure reduction in both treatment groups after 8 weeks. In comparison with the previous medications the proportion of patients with blood pressure reduction < 90 mm Hg diastolic was doubled in both treatment arms after 4 weeks: after initial dose of candesartan cilexetil 8 mg from 36.7% to 78.8%, after initial dose of candesartan cilexetil 16 mg from 43.9% to 81.1%. Clinically relevant side effects were not observed. CONCLUSION: Switching of antihypertensive monotherapy with ACE inhibitors, beta-blockers or calcium channel blockers to candesartan cilexetil 8 mg or 16 mg under ambulatory conditions is safe and equally well tolerated and effectively reduces blood pressure. PMID- 11402911 TI - [Benign metastasizing leiomyoma of the lung--a rare differential diagnosis of pulmonary space-occupying lesions]. AB - HISTORY AND ADMISSION FINDINGS: A 45-year-old woman was refferred for diagnosis of an accidentally found symptomless space-occupying lesions in the central part of the right lung. She had undergone a hysterectomy 4 years before and reported smoking 15 cigarettes daily since the age of 17 years. Physical examination was normal. INVESTIGATIONS: As primary bronchial carcinoma or metastasis to the lung was suspected she underwent a series of diagnostic tests: sonography, computed tomography (CT), gastroscopy, coloscopy, bronchoscopy, skeletal scintigraphy, gynaecological examination and various laboratory tests, none of which indicated a primary extrapulmonary tumour. CT-guided fine-needle biopsy then suggested benign metastasizing pulmonary leiomyoma (BMPL). TREATMENT AND COURSE: The largest of the tumours were surgically removed, confirming BMPL. Hormone receptors (for oestrogen, progesterone) having been demonstrated, progesterone treatment was initiated as prophylaxis against recurrences. CT 6 months later revealed new intrapulmonary foci. Administration of luteinizing hormone-releasing hormone analog to stop completely any oestrogen effect, and CT of the thorax 6 months later showed that both tumour numbers and their size had been reduced. The patient remained asymptomatic and the findings had not changed in the subsequent 12 months. CONCLUSION: BMPL is a rare cause of a space-occupying pulmonary lesion, predominantly affecting middle-aged women after hysterectomy for uterine myoma. The pathogenesis remains unclear, hormone-dependent tumour growth being discussed as a possible mechanism. Anti-oestrogen administration is the treatment of choice to achieve remission and effective prevention of recurrences. PMID- 11402912 TI - [Chronic anemia and abdominal pain as a sequela of lead poisoning]. AB - HISTORY AND ADMISSION FINDINGS: 3 years ago, a 63-year-old man underwent laparotomia due to clinical signs of acute abdominal pain. The anemia was treated with erythrocyte concentrates. Several weeks before the current admission to our outpatient department, the patient was again hospitalized because of acute abdominal pain and anemia. The patient was then transferred to the University clinic for further medical tests. INVESTIGATIONS: In addition to a normocytic anemia, a secondary porphyria was found. Further medical tests demonstrated lead poisoning. DIAGNOSIS, TREATMENT AND COURSE: Initially, the cause of the lead poisoning remained unclear because the patient had reported neither an occuppational nor an environmental exposure to lead. Finally, a prescription prepared ointment was evaluated, which the patient had applied to his feet and lips over the past 3 years. CONCLUSION: Lead poisoning can be found outside the realm of occupational exposure; therefore, the knowledge of lead poisoning is still important. PMID- 11402913 TI - [Repaglinide]. PMID- 11402914 TI - [Longitudinal melanonychia. Diagnosis, differential diagnosis and therapy]. PMID- 11402915 TI - [Statins as new therapeutic possibility in osteoporosis?]. PMID- 11402916 TI - [Disposition of the estate of a patient to the treating physician. Decision of the Karlsruhe high court 18 February 1999]. PMID- 11402917 TI - [Eradication therapy of Helicobacter pylori in antibiotic resistance]. PMID- 11402918 TI - [Therapy of angina pectoris: morphine or thalamonal?]. PMID- 11402919 TI - [Virus transmission by stab wounds]. PMID- 11402920 TI - [Polycythemia vera--current status of therapy]. PMID- 11402921 TI - [Felodipine in hypertensive patients with mild encephalopathies]. PMID- 11402922 TI - [Diabetes mellitus: the therapeutic challenge of the 21st century]. PMID- 11402923 TI - [Polymorphism of pro12Ala in peroxisome proliferator activated receptor gamma 2 (PPAgamma2): beta cell function and insulin sensitivity]. AB - BACKGROUND AND OBJECTIVE: The peroxisome proliferator-activated receptor isoform gamma (PPAR gamma) is a key regulator in lipid and glucose homoeostasis. A common polymorphism (Pro12Ala in PPAR gamma 2, prevalence ca. 25%) was shown to be associated with a decreased risk of type 2 diabetes. Generally, both beta-cell dysfunction and insulin resistance contribute to the development of type 2 diabetes. Therefore, the aim of the present study was to assess the mechanism by which the Ala allele of this polymorphism contributes to the reduced risk for type 2 diabetes. PATIENTS AND METHODS: We studied 51 subjects without (Pro/Pro) and 26 subjects with this polymorphisms (X/Ala) (both groups non-diabetic) by a modified hyperglycaemic clamp which permitted determination of both insulin secretion (in response to glucose, GLP-1 and arginine) and insulin sensitivity. RESULTS: None of the various phases of insulin secretion was significantly different between the 2 genotype groups (all p values > 0.13). In contrast, insulin sensitivity was significantly greater in X/Ala (0.19 +/- 0.03 U) compared to Pro/Pro (0.14 +/- 0.01 U, p = 0.04). In a two-dimensional assessment of insulin sensitivity and secretion, the homozygous alanine carriers appeared to have the most favourable constellation. CONCLUSION: These simultaneously obtained data for insulin secretion and sensitivity strongly suggest that the mechanism by which the Ala allele contributes to a risk reduction for type 2 diabetes most likely involves an increase in insulin sensitivity. PMID- 11402924 TI - [Costs of type 2 diabetes in Germany. Results of the CODE-2 study]. AB - BACKGROUND AND OBJECTIVE: Diabetes is an increasing and widespread chronic disease causing considerable costs for the health care system. In the CODE-2 Study (Costs of Diabetes in Europe-Type 2) the total expenses for type 2 diabetics in Germany were evaluated and analyzed for the first time. PATIENTS AND METHODS: The CODE-2-study has been performed in eight European countries. In the German arm of the study, medical, demographic, and economic data of 809 patients were obtained retrospectively for a one year period, using face-to-face interviews with 135 physicians. These results were projected for the overall population of type 2 diabetes patients in Germany. RESULTS: The annual costs caused by type 2 diabetes patients in Germany in 1998 amount to 31.4 billion DM. The majority of these costs (61%) were covered by statuatory and private health insurance. The annual expenses of the statuatory Health Insurance (SHI) for these patients amounted to 18.5 billion DM. These costs divided in 50% spent for inpatient treatment, 13% for ambulatory care, and 27% for medication. Diabetes medication (insulin, oral antidiabetic drugs) accounted for only 7% of total SHI costs. Only 26% of all diabetic patients were adjusted to HbA1c values < 6.5% according to the therapeutic targets of the European Diabetes policy group. 50% of the type 2 diabetic patients exhibited severe macro- and/or microvascular complications. The costs per patient--compared to the average expenses for SHI insured patients--increased with complication state from the 1.3-fold (no complications) up to the 4.1-fold (macro- and microvascular complications). CONCLUSIONS: The overall costs for patients with type 2 diabetes are higher than expected from previous estimates. Diabetes related complications and concomitant diseases are the predominant reasons for these high costs. Control of blood glucose is inadequate for the majority of diabetic patients. To prevent long-term complications, an optimized treatment of type 2 diabetes is imperative not only from a medical but also from a health economics point of view. PMID- 11402925 TI - [Spinal meningioma as differential diagnosis of diabetic polyneuropathy]. AB - HISTORY AND ADMISSION FINDINGS: A 70 year old woman had suffered from diabetes mellitus type 2 since she was 52. Three years before the surgery she had begun to experience weakness together with altered sensitivity in the right leg, which was regarded as having been caused by diabetic polyneuropathy. During the admission examination the level for algesia on the right-hand side was at about D 11, a distal paraparesis of the leg (3-4 degrees, Janda's classification), more intense on the right, hyperactive deep tendon reflexes, Babinski's reflex on both sides, and depressed abdominal cutaneous reflexes. The sensitivity to vibrations on the Malleolus medialis on both sides was 0/8. The patient could walk only with the help of a Rollator. INVESTIGATIONS: Over the three-year period following onset of symptoms the following tests were carried out: motor nerve conduction speeds of the N. tibialis and N. peronaeus, electromyogram of the N. tibialis anterior and the M. gastrocnemius, somatosensory evoked potentials (SSEP) of the N. tibialis, which indicated a lesion in the peripheral nerves or nerve roots. Cranial computed tomography (CCT), CT scan of the lumbar spine (L3-S1) and angiological investigation elicited no significant pathological findings. An MRI of the thoracal spine showed a vertebra-sized dorsal tumor pressing on the spinal cord from left to right. TREATMENT AND COURSE: By means of microsurgery the spinal tumor was completely removed. Suspected meningeoma was confirmed by histological analysis. During the post-surgical period, the incomplete paraplegia quickly regressed, and 7 weeks after the removal of the spinal meningeoma the patient was able to climb stairs. CONCLUSION: In case of slowly-developing paresis of the legs in diabetic patients, diabetic polyneuropathy should not be diagnosed without careful consideration, and rare spinal tumors should be considered as part of the differential diagnosis, especially if the blood glucose level is normal, and intensive physiotherapy brings no improvement in the patient's condition. PMID- 11402926 TI - [Prediction and prevention of type 1 diabetes]. PMID- 11402927 TI - [Clinical effects and pharmacodynamics of insulin analogs lispro, aspart and glargin]. PMID- 11402928 TI - [Value of glitazones in therapy of type 2 diabetes: pro]. PMID- 11402929 TI - [Use of glitazones in type 2 diabetes: a critical position]. PMID- 11402930 TI - [Metformin and anesthesia--how great is the risk of lactic acidosis]. PMID- 11402931 TI - [Significance of HbA1c]. PMID- 11402932 TI - [Methotrexate]. PMID- 11402933 TI - [Depression in HIV positive women]. PMID- 11402934 TI - [Diuretics in the treatment of arterial hypertension]. AB - BACKGROUND: Diuretic compounds, which comprise the basis of any cardiovascular prevention in subjects with hypertension, have never been studied in terms of clinical pharmacology according to the classical and well-established rules used nowadays with other anti-hypertensive agents. FINDINGS IN THE LITERATURE: Because edema was the first historical indication for diuretic compounds, very high doses were initially given for drug treatment of chronic hypertension. It was only 10 15 years ago that lower doses were first used due to the effectiveness of treatment in old subjects with hypertension and to the development of fixed associations with either converting enzyme inhibitors or beta-blocking agents. However, only a very small number of pathophysiological studies have been conducted in parallel, particularly to determine the effects on vessels and the endothelium. CONCLUSION: Diuretics are the most effective and the least expensive drugs for cardiovascular prevention of hypertension but the basic mechanisms of their action remains to be evaluated. PMID- 11402935 TI - [VO2 max: technique and practical importance]. AB - NORMAL SITUATION: Ergonometric spirometric measurement of O2 consumption and CO2 production (VO2max) can be used to assess adaptation of the oxygen transportation system to exercise. In the normal situation, the capacity to sustain exertion in everyday physical activities depends directly on the capacity of this system to adapt, i.e. the level of physical training. PATHOLOGICAL CONDITIONS: In diseased patients, this test can measure the degree of non-adaptation and the efficacy of compensatory mechanism of the oxygen transport system. It can be used as a pathophysiological diagnostic tool for following divers disease states, particularly heart conditions. Spirometry is a key test used in all cardiology centers to assess the degree of heart failure. PMID- 11402936 TI - [Cardiac manifestations of sickle cell anemia]. AB - HEMOGLOBINS S AND C: Drepanocytosis, the occurrence of sickle cells (drepanocytes) in the blood, is an inherited condition. Electrophoresis demonstrates hemoglobin SS in homozygous subjects who present the typical clinical features of severe hemolytic sickle-cell anemia. Heterozygous subjects have sickle-cell anemia trait, an asymptomatic condition associated with a 50% hemoglobin S and 50% hemoglobin C at electrophoresis. Hemoglobin S and C are transmitted by Mendelian inheritance. CARDIAC DISORDERS: Well-known, cardiac disorders occur in more than 82% of homozygous subjects while only 2% of heterozygous subjects are affected. Heart murmur, radiological cardiomegaly, or eletrocardiographic anomalies are often the only signs. There is a risk of fatal heart failure in children and neonates. Acute rheumatic fever or infectious endocarditis, particularly due to pneumococcal or Haemophilus influenzae infection, may trigger heart failure. CARDIAC ANOMALIES: Patients with sickle cell anemia can develop an "anemic heart" expressed by an elevated cardiac output and systemic ejection volume at rest and a fall in arteriolar peripheral resistance. Patients who develop cor pulmonae have an elevated pulmonary pressure at exercise and experience venous occlusive events with a progressive reduction in the pulmonary vascular bed and development of a left-right shunt. Myocardiopathy leads to left ventricular dysfunction contrasting with the dilated right heart seen at echocardiography and rare cases of transmural infarction. PMID- 11402937 TI - [Intracoronary radiotherapy (brachytherapy) for treatment of restenosis]. AB - RESTENOSIS AFTER CORONARY ANGIOPLASTY: Restenosis after coronary angioplasty remains a common clinical problem. Even after the advent of stenting, the rate of restenosis is still to the order of 20% or even higher in certain patient subgroups. Treatment is difficult, particularly in case of diffuse restenosis within stent lumens. POTENTIAL CONTRIBUTION OF BRACHYTHERAPY: After stent implantation, restenosis is generally due to excessive intimal proliferation producing a "benign tumor" formation that progressively obstructs the arterial lumen. Endocoronary curietherapy has been proposed for its "antiproliferative" effect. SYSTEMS USED: Two radiation sources are used: gamma emitters with powerful tissue penetration properties, and beta emitters which exhibit less tissue penetration. Radioprotection is a greater problem with gamma emitters but dosimetry is more difficult to control with beta emitters. In clinical practice, the sources are placed close to the target site via conventional percutaneous catheterization and are employed as a complement to standard angioplasty used to destroy the stenosis. Endocoronary brachytherapy requires close collaboration between the interventional cardiologist and the radiotherapist. CLINICAL TRIALS: After the preliminary feasibility studies, several randomized trials have assessed the contribution of brachytherapy in the prevention of restenosis. Using gamma emitters (192Ir), the SCRIPPS, WRIST and GAMMA-1 studies demonstrated good immediate safety and a real effect on restenosis with 50% reduction. Inversely, groups of treated patients have a higher rate of severe complications (death and infarction) related to late acute coronary thrombosis after interruption of antiplatelet therapy combining ticlopidin and aspirin. These complications result from the frequent introduction of stents during angioplasty procedures and by delayed endothelialization of the stents leaving a starting point for thrombosis formation longer than usual. The same type of complications have been observed with beta brachytherapy in the PREVENT trial, while in the START trial that used the 90Sr/y source in 476 patients, severe events were not more frequent in the irradiated group. In the latest trial, the frequency of new stent implantations was lower (21%) and antiplatelet treatment was maintained longer. Finally, the rate of restenosis was significantly lower (29% versus 45%). PRACTICAL APPLICATIONS: Current data confirm the efficacy of endocoronary brachytherapy to limit intimal proliferation after angioplasty and reduce the rate of restenosis. The question of safety remains open, with the potential risk of late coronary thrombosis; no long-term data (more than 5 years) are available. It is undoubtedly too early to propose routine use of brachytherapy, excepting perhaps cases where interventional cardiology raises particularly difficult problems, for example in case of diffuse restenosis within an endoprosthesis. PMID- 11402938 TI - [Acute respiratory distress syndrome after antineoplastic chemotherapy. Probable role of gemcitabine]. AB - BACKGROUND: A 4-week interval between radiotherapy and gemcitabin chemotherapy is recommended due to the risk of severe radiosensitization. Gemcitabin can also have severe lung toxicity late after or without prior radiotherapy. CASE REPORT: A patient was treated with thoracic radiotherapy for non-small-cell lung cancer. Five weeks later gemcitabin was given. A few days after the second gemcitabin cycle the patient developed severe respiratory distress. The clinical course was rapidly fatal despite corticosteroid therapy. DISCUSSION: About 20 cases of severe lung toxicity due to gemcitabin have been reported in the literature, occurring late after radiotherapy or without radiotherapy. Corticosteroid therapy, whether given for prevention or cure, is not always effective. Four of these cases had a fatal outcome. The development of brief mild episodes of dyspnea is considered to be common after delivery of gemcitabin. If unexplained dyspnea persists for more than a few hours, severe lung toxicity is highly likely and gemcitabin should be interrupted. PMID- 11402939 TI - [Homonymous lateral hemianopsia revealing cerebral thrombophlebitis. Role of a deficiency of protein S induced by l-asparaginase]. AB - BACKGROUND: Cerebral thrombosis associated with protein S deficiency is very rare and is mainly related to hereditary form of protein S deficiency. CASE REPORT: A 19-year-old girl with acute lymphoblastic leukemia presented hemianopsy within a few days after the first administration of L-asparaginase. Magnetic resonance of the brain showed a cortical infarct. A marked decrease of the level of protein S was documented. Few days later, the patient was free of symptoms and protein S level was restored to the normal suggesting that the cerebral thrombosis was caused by transient protein S deficiency induced by L-asparaginase administration. DISCUSSION: Patients with neurological complication caused by L asparaginase should be tested for protein S and other anticoagulant deficiencies. PMID- 11402940 TI - [Superficial injections of anti-inflammatory drugs and severe infectious complications]. PMID- 11402941 TI - [Impaired visual acuity in Mediterranean boutonneuse fever]. PMID- 11402942 TI - [ABO-incompatible heart transplantation in infants]. PMID- 11402943 TI - [Medical record, source of conflict]. AB - Patients' right to information is strictly regulated by the French Code of Public Health that establishes the legal framework for delivering medical information. The Code of Medical Deontology also contains several articles concerning patients' rights, medical information, and medical secret. The patient's right to information must not be confounded with medical file property rights which is an ambiguous notion that has not yet acquired a perfectly acceptable legal definition satisfying all concerned. Currently, the physician, designated by the patient, is the only person legally authorized to deliver to that patient the desired information. PMID- 11402944 TI - [Implantable defibrillators. Sudden death: high risk]. AB - INCIDENCE: The incidence of sudden death remains high. The basic causes are related to ischemic heart disease and heart failure. Automatic defibrillation is the best treatment, as first conceived and implanted by Mirovski twenty years ago. MARKERS OF RISK OF SUDDEN DEATH: Left ventricular dysfunction, functional class and spontaneous and induced ventricular hyperexcitability identify a high risk population. THERAPEUTIC APPROACH: Drug treatments used in the post infarction period or for heart failure (beta-blockers, converting enzyme inhibitors and amiodarone) have demonstrated a certain efficacy in preventing sudden death. However, different therapeutic trials comparing the efficacy of defibrillators and drug treatments have demonstrated the 20 to 46% superiority of defibrillation in terms of reduced mortality. PMID- 11402945 TI - [Implantable defibrillators. Role of the defibrillator in treatment and prevention of sudden death]. AB - PROVEN EFFICACY: Several studies have demonstrated that implantable defibrillators improve survival in patients with a very high risk of sudden death. INDICATIONS FOR PRIMARY PREVENTION: The most obvious indication is for young patients with severe recurrent rhythm disorders and generally good left ventricular function. However, most of the candidates have severe left ventricular dysfunction due to post infarction ischemia or dilated cardiomyopathy who develop syncopal or hemodynamically poorly tolerated ventricular tachycardia or who have survived a first episode of sudden death. PROPHYLACTIC INDICATIONS: Automatic defibrillation should be proposed for patients with factors of risk of sudden death such as left ventricular dysfunction or infarction sequelae. PMID- 11402946 TI - [Clinical experimentation with contrast media in Italy]. PMID- 11402947 TI - [Pulse inversion ultrasonography with ultrasonography contrast media (levovist) in the evaluation of hepatic metastasis]. AB - PURPOSE: To evaluate if Pulse Inversion Harmonic Imaging with contrast agent injection (Levovist) is able to determine an increase of echogenicity in normal hepatic parenchyma, and to compare its capability to detect metastatic focal lesions with that of standard US and spiral CT. To define and standardize the technical and methodological aspects of this new technique. MATERIAL AND METHODS: A selected group of 72 patients (42 males and 30 females) with clinical and instrumental suspect of hepatic metastatic lesions was included in the prospective study. Each patient was examined by conventional ultrasound (US), Pulse Inversion Harmonic Imaging (PI) and spiral-CT (spCT). US examination was performed using an HDI 5000 (ATL, Bothell, USA) equipped with a broadband probe (5.0-2.0 MHz). 2.5 g Levovist (Schering AG-Berlin) was administered intravenously, at concentration of 300 mg/ml and a flow rate of 7 ml/min. Video clips obtained with the acquired images before and after contrast medium administration were transferred to a magnet optic disk unit. Examinations were evaluated by an experienced radiologist blindly on a workstation that allowed a qualitative-quantitative analysis. SpCT images were evaluated separately by another experienced radiologist. US images were evaluated qualitatively (number of lesions, site of lesion, diameter of the smallest lesion detectable) and quantitatively (increase in parenchymal echogenicity 20", 40", 60" and 5' after the injection of contrast agent). RESULTS: In all cases examined, the echogenicity of normal hepatic parenchyma increased after contrast agent administration, reaching a peak of about 250% (1DS) at 60 s and a decreasing gradually in 5 minutes. Conventional US detected 195 focal lesions, CT 231, and US with pulse inversion technique and contrast agent (US-PI) 287. Mean differences among the number of lesions detected by the different techniques per each patient were also calculated. Wilcoxon Signed Rank Test showed a statistically significant difference between US-PI and US (p < .0001), CT and US (p = .0052) and US-PI and CT (p = .0121). US-PI detected the smallest lesions, which went undetected by the CT and conventional US examinations. DISCUSSION AND CONCLUSIONS: In 10 cases (14.3%) contrast enhanced US-PI demonstrated the presence of a number of lesions greater than 5 (diffuse metastatization and inoperability) in comparison to that detected by spCT). The greater echogenicity of normal hepatic parenchyma using pulse inversion technique after Levovist infusion allowed to better demonstrate hepatic metastases. This technique showed a higher identification rate of small lesions in comparison to basal examination and to spiral CT. Contrast enhanced US-PI demonstrated a remarkable increase in echogenicity of hepatic parenchyma in portal phase. The technique significantly improves the detection of focal lesions allowing visualization of smaller lesions compared to CT and US. These results indicate that the technique could be used in the staging of liver metastasis. However, large multicentric trials are required to validate US-PI and better define its role in the work-up of patients with neoplastic disorder. PMID- 11402948 TI - [Study of the liver and the portal venous system with digital rotational angiography]. AB - PURPOSE: Digital subtraction angiography is of common clinical use for the evaluation of vascular disease. The rotation of the X-ray tube around the patient's body during contrast medium injection was first proposed in the early seventies in the neuroradiologic field; only recently it has been applied to the evaluation of vascular structures and abdominal parenchymatous organs. We investigated the potential clinical value of digital rotational subtraction angiography in the evaluation of the hepatic arteries and of the portal venous system. MATERIALS AND METHODS: Digital rotational subtraction angiography was performed in 46 patients (34 males and 12 females), mean age 59.3 years (range: 43-72). All the patients underwent digital rotational subtraction angiography after ultrasonographic, CT and/or MRI imaging for evaluation of hepatocellular carcinoma (31 patients) prior to trans-arterial chemoembolization, hepatic metastases from gastrointestinal cancer (9 patients) and pre-surgical study in portal hypertension (6 patients). Digital rotational subtraction angiography was performed using the following technical parameters: a maximum frame rate of 10 views per second, a 1024 x 1024 matrix, a rotation time of 5 seconds, a rotational arch of 90 degrees with a speed of 30 degrees/second. Digital rotational subtraction angiography of the liver was carried out after positioning of a Cobra angiographic catheter in the proper hepatic artery or in the left or right hepatic artery, and subsequent injection of 20-30 ml on contrast medium at a flow rate of 4-7 ml/sec. Conversely, in the portal study the catheter was placed in the splenic or superior mesenteric artery and contrast was administered at 10 ml/sec for an amount of 40-60 ml. Conventional, non-rotational angiography was always obtained with the same catheter and less contrast medium (15-25 at 4-7 ml/sec in the hepatic study, 25-40 ml at 5-7 ml/sec in the portal study). We have evaluated the diagnostic quality (rated as equal, superior or inferior) and the presence of image noise of digital rotational subtraction angiography when compared to digital non-rotational subtraction angiography. We also evaluated the tolerability and the mean time to perform the examination. RESULTS: Compared to non-rotational digital subtraction angiography, the diagnostic quality of digital rotational subtraction angiography was superior in 26 cases, equal in 20 and never inferior: these results are particularly evident in cine-mode. Diagnostic efficacy was similar in the arterial phase and generally better in the venous phase. Image noise was always perceptible, mostly in lateral and oblique views and is related to the patient's size. Noise especially hindered evaluation of the portal venous phase. Digital rotational subtraction angiography was well tolerated by all patients, although its most significative drawback was the prolonged apnea time required (about 8 seconds per single rotation) which can sometimes be difficult for elderly patients. Examination duration is about 5 to 10 minutes. Contrast medium doses required never exceeded 20-60 ml. DISCUSSION: Current evaluation of an hepatic lesion requires injections and multiple views to fully delineate arterial anatomy. This requires the radiologist to create a mental 3-D rendering based upon a 2-D view, obtained on the basis of the radiologist's experience. Rotational angiography, when reviewed in cine-loop, allows a better 3-D rendering than conventional angiography, increasing the advantages of the multiple views obtained from a single angiographic run and allowing an exact imaging of the course and direction of the hepatic arterial branching, making selective catheterization during trans-arterial chemoembolization or other interventional procedures easier. Magnification further improves the evaluation of a mass and of the arterial tree. In hepatic surgery, the most important problem is the precise knowledge of the segment involved and the position of the lesion inside the segment, since a mass in the middle of the segment requires a segmentectomy while a mass near the borders is treated by a larger resection. Digital rotational subtraction angiography improves the visibility of vascular anatomy, allows a better knowledge of hepatic artery branches and improves the angiographic investigation of the liver, permitting a correct spatial assessment. CONCLUSION: Digital rotational subtraction angiography is a useful tool for the evaluation of the liver as well as for transarterial chemoembolization or other interventional procedures. Images are obtained during a single contrast injection, with a better 3-D rendering of the hepatic artery and the portal venous system: no other method provides as complete a visualization of liver vascular anatomy after a single injection of contrast medium in one examination series. (ABSTRACT TRUNCATED) PMID- 11402949 TI - [Role of videofluorography in the study of esophageal motility disorders]. AB - PURPOSE: To repropose the importance of videofluoroscopy in the study of esophageal motor disorders, comparing the radiologic and manometric results; the manometric results are considered the reference parameters. MATERIAL AND METHODS: From 1996 to 1999, 76 patients (42 males and 34 females), were studied first using manometry and then videofluoroscopy. The patients had symptoms like dysphagia, thoracic pain or both. The manometric study was performed with a perfusional system equipped with 6 tips (4 radial for the study of the esophageal sphincters and 2 placed longitudinally for the study of the esophageal peristalsis). With the patient in a supine position we analysed 5-10 deglutitions with 5 ml water bolus at 20-25 degrees C, administered using a graduated syringe. The radiologic study was performed with a remote-control digital television system, connected to a video recorder. Three 7.5 ml bolues of high density barium suspension (250% weight/volume) were injected orally in the upright position and other three were injected in the prone position following the passage from the oral cavity to the stomach. RESULTS: The comparison of the manometric and videofluoroscopic results suggests that the total sensitivity of the radiological study in the detection of esophageal motor disorders was 92%. In particular dynamic radiologic investigation diagnosed the normal esophageal functionality in 100% of the cases, nonspecific esophageal motility disorders in 89.6%, diffuse esophageal spasm in 100% of the cases, the presence of achalasia in 90%, whereas "nutcraker esophagus" only in 50%. Videofluoroscopy therefore showed high sensitivity in four groups of the five considered. It has some limitations in the diagnosis of initial achalasia, and is not sufficiently sensitive in the diagnosis of "nutcracker esophagus". CONCLUSIONS: Videofluoroscopy is a simple method which presents high sensitivity and specificity in the detection of motility disorders of the esophagus and could therefore be proposed as the first diagnostic method in patients with specific symptoms. PMID- 11402950 TI - [Lung base disease after orthotopic liver transplantation. Functional ultrasonography of the diaphragm and correlated alterations]. AB - PURPOSE: To demonstrate the greater accuracy of B- and M-probe ultrasound (US) compared to traditional examinations in the evaluation of diseases of the lung base, a frequent localisation of pathology in patients undergoing orthotopic liver transplantation (OLT). MATERIAL AND METHODS: Twenty-one patients, 13 males and 8 females aged 24 to 63, awaiting OLT were examined using the three modalities. B-mode US was performed as a preliminary study to identify the districts of interest and the profile of the diaphragm wall, searching for any related alterations. This was followed by M-mode US, with an approach along the left and right posterior axillary lines during spontaneous and forced maximal expiration, to calculate and document the curve representing diaphragm mobility. All patients were also studied pre- and postoperatively by standard chest X-ray double projections. The parameters evaluated by US were diaphragmatic inspiratory slant and diaphragm range while the standard chest X-ray was used to assess hypoventilation, diaphragm range and pleural effusion. RESULTS: For each parameter considered we obtained the following results: presence or absence of pleural effusion (sensitivity: 100% with US vs 64% with chest X-ray) and diaphragmatic hypomobility with related hypoventilatory phenomena (sensitivity: 85% with US, with 15% false negatives). In 15 cases the chest X-ray revealed a clear elevation of the diaphragm, a finding supported by US in 11 cases. In 7 cases US showed a reduction in the diaphragm range curve without, however, any radiological evidence of any ventilatory dysfunction of the lung base and/or elevation of the corresponding hemidiaphragm. CONCLUSION: Our results suggest that radiology, B-mode US and color Doppler US, which are widely used for monitoring OLT patients, can be usefully integrated by M-mode US to evaluate diaphragmatic mobility both pre- and post-operatively. This method is fast, easy to use and widely available. PMID- 11402951 TI - [Vertebral morphometry: evaluation of osteoporosis-caused fractures]. AB - PURPOSE: To compare visual reading of spine radiographs and quantitative morphometric approach for assessing the prevalence of vertebral fractures in postmenopausal osteoporosis. MATERIAL AND METHODS: In 473 postmenopausal women afferent to our Centre of Osteoporosis under-went lateral thoracic and lumbar spine radiograph to identify vertebral fractures and dual energy X-ray absorptiometry (DXA) to measure bone mineral density (BMD) of the lumbar spine (L1-L4). Osteoporosis was defined according to the World Health Organization (WHO) guidelines. To identify vertebral fractures the radiographs were visually analyzed by two radiologists; a woman was judged as fractured only if both readers independently found at least one vertebral fracture on her films. Then the spine radiographs were digitized by means of a scanner to perform quantitative vertebral morphometry (QVM) using specific software. An expert operator manually located the calipers on the vertebral bodies from T4 to L5 and the computer automatically calculated the anterior, middle and posterior vertebral heights and their ratios. A vertebral fracture was defined by morphometry as a reduction by at least 20%, with an absolute decrease of at least 4 mm, in one of three height ratios of any vertebral body compared to the corresponding reference ratio for fertile women. RESULTS: Visual reading by radiologists detected 9.5% (45/473) women with vertebral fractures and QVM detected 13.7% (65/473) with statistical significance (p < 0.001). In the 75-80 years age group the prevalence of vertebral fractures reaches the maximum value, 26.3% by visual reading and 36.8% by QVM. Among fractured women, 34 were osteoporotic by DXA; 11 women found fractured by visual reading and 21 by QVM were osteopenic women, with bone mineral densities between -1 and -2.5 SD of the T-score. CONCLUSION: This study showed that quantitative assessment of spine radiographs by vertebral morphometry is an objective method that allows to identify a larger number of vertebral fractures compared to visual inspection. This is very important not only for epidemiological studies, but also for clinical use because a previous vertebral fracture increases the risk of subsequent fractures significantly. Therefore, to improve the risk assessment of vertebral fractures for osteoporotic patients it is necessary to combine the use of QVM and BMD. PMID- 11402952 TI - Dynamic contrast enhanced magnetic resonance imaging subtraction in evaluating osteosarcoma response to chemotherapy. AB - PURPOSE: To evaluate the results of a new technique of dynamic contrast enhanced Magnetic Resonance (MR) imaging subtraction in the assessment of osteosarcoma response to chemotherapy. METHODS: 24 patients with high grade osteosarcoma, treated with preoperative neo-adjuvant chemotherapy, underwent MR at high field strength (1.5 T). Both unenhanced conventional SE T1- and T2-weighted sequences in the coronal and axial plane and dynamic Gd-DTPA-enhanced SE T1-weighted sequences in the coronal plane were performed. Image postprocessing included subtraction of unenhanced image from enhanced images (arbitrary called "angiographic subtraction") and subtraction of each enhanced image from the last enhanced image (arbitrary called "pathologic area" subtraction). The early enhancing areas detected in the angiographic subtraction and the pathologic areas detected in the pathologic area subtraction were correlated with histopathological findings on histological macrosections obtained from the resected specimen. The sensitivity, specificity, accuracy, positive and negative predictive value of both the subtraction techniques were calculated. RESULTS: The early enhancing areas of angiographic subtraction were related not only to viable tumor but also to the host reactions such as flogosis and granulation tissue. The pathologic areas detected at the pathologic area subtraction correlated in most cases with viable tumor, while in 3 cases they did not correspond to viable tumor tissue and in 1 case a small area of residual viable tumor was missed. In assessing response to chemotherapy, pathologic areas subtraction had an accuracy of 95% (specificity: 100%, sensitivity: 93%, PPV: 100%, NPV: 88%), whereas angiographic subtraction had an accuracy of 79% (specificity: 37%, sensitivity: 100%, PPV: 76%, NPV: 100%). CONCLUSIONS: Pathologic area subtraction may be a useful technique for assessing the response of osteosarcoma to chemotherapy and for detecting residual viable tumor tissue. PMID- 11402953 TI - [Anatomic and radiological observations of th bifid mandibular condyle]. AB - PURPOSE: To determine the morphology and the frequency of bifid mandibular condyle (BMC) in anatomic material and to study its diagnosis in living persons. MATERIAL AND METHODS: The morphology and the frequency of BMC were studied on a sample of 210 intact mandibular condyles belonging to a collection of 200 dry male adult skulls held by the Institute of Anatomy of University of Bologna. Diagnostic imaging was verified retrospectively on 6 cases of BMC observed in living subject. RESULTS: In the anatomic material, we found 29/210 mandibular condyles with the articular surface simply parted in two (13.80%) and 1 condyle consisting of two separate articular heads (0.47%) at different levels and without the neck. Therefore BMC with separate neck is very rare. In living persons diagnosis was established by radiology using orthopantomography and demonstrated 2 cases of bilateral BMC (one dysplasic and the other traumatic) and 4 of unilateral BMC. DISCUSSION AND CONCLUSIONS: We believe that primitive BMC is a consequence of a dysplasic process of the condyle cartilage at the early stages of its development, while secondary traumatic BMC is caused by bone remodelling after articular trauma or repeated microtraumas from malocclusion. In dysplasic BMC, as evidenced by the observation made on a living subject, a distinction should be made between the complete type in which the accessory articular head is separate from the principal head, at a different level and with its own separate neck, and the other, incomplete types. Orthopantomography, which proved to be a valid diagnostic tool, must be integrated with CT and MR if surgery is planned. PMID- 11402954 TI - [Embolization of the uterine artery in the treatment of uterine myoma]. AB - PURPOSE: To propose uterine myoma embolization as an alternative to myomectomy or hysterectomy in the treatment of symptomatic myomas; to evaluate the efficacy of the procedure in terms of clinical outcome, adopting all procedural and technical precautions to ensure minimal X-ray exposure and preserve reproductive potential. MATERIAL AND METHODS: Between April 1998 and February 2000, 26 patients, age range 32-54 years (mean 41 years), underwent uterine arterial embolization for menorrhagia, pelvic pain, and sensation of mass and pressure. Inclusion criteria were: single myomas, intramural localization and rich vascolarization of the lesion. Dose to patient was obtained by placing a thermoluminescent dosimeter (Harshaw, Solon, Ohio) both placed in posterior fornix of the vagina and on the skin at the beam entrance site. The procedure was performed under peridural anesthesia; polyvinil alcohol particles 355-500 mu (Contour) (Target Therapeutics, Boston Scientific Corporation, Fremont CA, USA) were employed as embolic agent. The uterine arteries were incannulated with a 5F (Glidecath, Terumo, Europe NV, Belgium) and successively 3F coaxial microcatheter (Target, Boston Scientific Corporation, Fremont CA, USA); the embolic material was injected as distally as possible. Color Power Doppler Ultrasound follow-up before and after i.v. contrast media administration (Levovist SHU 508 A, Shering, Berlin, Germany) was carried out at 15 days, at 1, at 3, at 6 months, and at 1 year from embolization. Pre-procedural evaluation and follow-up at 1 year was performed by MRI using T1 and T2 weighted images before and after Gadolinium (GdDTPA Shering, Berlin, Germany) administration. RESULTS: The technical success of the interventional procedure was 100% (26/26 cases). The mean fluoroscopy time was 20 minutes, and the mean number of angiographic exposures was 10. The mean estimated ovarian dose was 18.75 cGy and the mean adsorbed skin dose was 126.71 cGy. The imaging follow-up showed a 55% reduction of myoma volume at 6 months and a 75% reduction at 1 year. All patients reported a marked decrease in symptoms. No major complications were observed. The appearance of pelvic pain in the 24-48 hours after the procedure required sedation by analgesic pump; transitorial amenorrhea was observed in 3 patients. As for term complications, 2 patients have eliminated necrotic material through the vagina four weeks after procedure. The patients reported great satisfaction with the procedure. DISCUSSION: Many treatment options are currently available for symptomatic uterine myomas. One is surgical myomectomy which is associated with increased blood loss, pain and post operative morbidity and requires an additional surgical procedure for fibroma recurrence in 20-25% of patients. Another alternative treatment is hormonal therapy, which drammatically improves symptoms and reduces fibroid size although leiomyomas regrow to their original size within a few months of discontinuing treatment. Uterine embolization is a relatively new treatment for uterine fibroids that can be considered as an alternative to surgical and medical procedures. The radiation exposure adsorbed by the patient is reduced by using pulsed fluoroscopy and taking all the precautionary measures required to minimize the dose. CONCLUSIONS: The technical success, the patient' satisfation, the short hospitalization time and preservation of fertility confer to uterine artery embolization the role of a new alternative therapy for the treatment of symptomatic uterine myomas. PMID- 11402955 TI - [Percutaneous treatment of varicocele. 13-year experience with the transbrachial approach]. AB - PURPOSE: To report our experience using the transbrachial approach, which is easily accepted by the patient, in the treatment of varicocele. MATERIAL AND METHODS: Between January 1986 and December 1998, 1490 patients with clinical or subclinical varicocele, but with seminal fluid alterations, underwent spermatic phlebography using the transbrachial approach. Since 1991 the procedure has also been adopted at the Unit of Pediatric Surgery of our hospital, which proposes it as a first choice treatment in adolescents with varicocele. The procedure consists in accessing the basilic vein at the elbow level percutaneously and using a hydrophile guidewire and multipurpose angiographic catheter to reach and catheterise the spermatic vein responsible for the varicocele. During the first years, we used sclerotherapy alone; subsequently, if the varicocele recurred or if the reflux was refractory to sclerotherapy or if the veins were large we adopted vein embolisation. Follow-up was one year and consisted of testicular ultrasound, Doppler flowmetry and/or color Doppler ultrasound at one, six and twelve months after the procedure. Patients were considered restored if they were free of symptoms, showed no venous reflux and/or had normal seminal fluid parameters and improved if they were free of symptoms but still presented venous reflux. Varicocele was considered persistent if the procedure failed to produce any beneficial effects, and recurrent if, although absent at the first follow up, it reappeared after the fifth month. RESULTS: We found 1296 (86.9%) cases of left varicocele, 25 of right varicocele and 169 (11.3%) of bilateral varicocele. In all cases, the symptoms disappeared after the percutaneous procedure. Duration of radioscopy was reduced to 3.5'; the procedure lasted 90' for the monolateral varicoceles and 120' for the bilateral forms. 313 diagnostic procedures were performed (20.7%). The procedure could not be completed in 104 patients (6.8%) due to basilic vein spasms, difficulties encountered in catheterizing the spermatic vein and, particularly in pediatric patients, anatomic variations. A total of 1195 (79.2%) procedures were completed: sclerotherapy alone in 642 patients and sclerotherapy followed by scleroembolisation in 527. Sclerotherapy alone was sufficient to restore 524 patients (86.6%), while 384 (78.5%) required scleroembolization. A small number of patients underwent scleroembolization alone, which brings the success rate for the two procedures to 82% and 84%, respectively. No serious side-effects were noted. DISCUSSION: The transbrachial approach in spermatic phlebography has proved to be a safe and effective technique for the treatment of both monolateral and bilateral varicocele. Furthermore, the procedure is well accepted by patients and can be performed in a day-care setting. In some cases, we only obtained partial results because of the large caliber of the spermatic vein; in other cases, we were unable to complete the procedure due to anatomic variations or to the spasm of the basilic vein. CONCLUSION: The safety and effectiveness of this procedure make it a valid alternative to traditional surgery, that should be considered as a possible first choice treatment for varicocele in adolescents. PMID- 11402956 TI - [Personal experience with computerized tomography of shock and death in emergency situations]. AB - PURPOSE: The wide availability of CT machines in hospitals specialising in emergency care has made it possible to observe CT features of severe shock and death of the patient during the CT examination. Unique radiological signs can be evaluated that testify to the state of shock or indicate arrest of the contrast medium in the cardiovascular system and parenchymatous organs, with different hemodynamic features from those found in living persons. This paper aims to report our personal experience with the role of CT in documenting states of shock or death. Our series, which is relatively large considering the infrequency of cases, seems to be of interest as it is one of the few to describe these rare CT findings, which have ethical implications for all healthcare providers and the field of diagnostic imaging. MATERIAL AND METHODS: In the last 5 years we have observed 16 cases (12 males and 4 females; mean age 41 years; range 17-79) of death during CT examinations. Of these, 9 were severely polytraumatized patients, 2 had ruptured aneurysms of the abdominal aorta, and one had a cardiac arrest due to cardiac infarction. Death occurred in 3 patients with head trauma, in whom the CT examination had shown signs of inoperability. RESULTS: The most frequent CT signs of death observed in the patients who died of polytrauma were contusive hemorrhagic foci in the lung associated with pneumomediastinum and hemothorax (9 cases), diffuse subcutaneous emphysema of the chest wall and pneumothorax (8 cases). CT revealed bilateral costal fractures and areas of pulmonary subatelectasia in 8 cases, and rupture of the diaphragm in one. Multiple lacerations of the liver and spleen with marked hemoperitoneum, incompatible with survival, were observed in 3 cases. Vertebral traumatic alterations were present in 2 cases. In 5 cases, the last CT scans of the heart and hepatic cupula revealed massive enhancement of contrast medium within the heart and great vessels due to contrast-medium hypostasis. The diaphragm was hypotonic, an indirect sign of death, with the left hemicardium and hemidiaphragm in a lower position than found in living persons; the abdominal aorta had a reduced caliber. DISCUSSION AND CONCLUSIONS: Such CT findings will probably become increasingly frequent in the future, because the need to avoid exploratory laparotomies or thoracotomies and unnecessary neurosurgical procedures will entail a greater demand for highly selective diagnostic examinations to be carried out on critical patients. This tendency is matched by the technological advances in diagnostic imaging, and particularly the availability of fast CT and spiral CT machines that give, in only a few seconds, an accurate diagnostic and clinical picture, on which immediate therapeutic decisions can be made. PMID- 11402957 TI - [Role of computerized tomography in the diagnosis of peritoneo-intestinal lesions resulting from closed trauma. Experience at 2 emergency departments]. AB - INTRODUCTION: Small and large bowel mesenteric injuries from blunt abdominal trauma are rare and often difficult to diagnose. Computed Tomography used in cases of blunt abdominal trauma has been found sensitive in detection of bowel and mesenteric injuries and discrimination of operable from nonoperable candidates. PURPOSE: A retrospective study of the CT examination of 24 patients, with surgically confirmed bowel and mesenteric injuries, was performed. Our goal was to evaluate the various CT signs of blunt bowel and mesenteric injury and the related frequency. MATERIAL AND METHODS: Our series includes 24 patients, 16 of them (first group) were investigated with CT at Cardarelli Hospital, Naples, while the remaining 8 (second group) at University of Bari. Patients of the first group, 11 men and 5 women, age ranging from 18 to 77 years, were submitted to a conventional abdominal CT performed after i.v. administration of contrast media. Patients of the second group, 7 men and 1 woman, age ranging from 4 to 81 years, were submitted to helical CT performed with the following parameters: 10 mm slice thickness, 5-mm contiguous intervals from the level of the diaphragm to the pubic bone, pitch 1.5. Helical CT was performed in all cases before and after i.v. administration of contrast material injected at a rate of 3.5 ml/sec. A scanning delay of 40 seconds after the beginning of contrast injection was routinely used. In all patients the following CT signs were retrospectively searched on: peritoneal or retroperitoneal fluid, mesenteric hematoma, hazy streaky changes in mesenteric fat, high-density clot (sentinel clot) adjacent to the involved bowel, pneumoperitoneum, retropneumoperitoneum, extravasation of intravenous contrast material, bowel wall thickening. RESULTS: In the first group the following CT signs were observed: mesenteric hematoma (87.5%), hazy streaky changes in mesenteric fat (56.25%), peritoneal or retroperitoneal fluid (37.5%), sentinel clot (25%), bowel wall thickening (18.75%), extravasation of intravenous contrast material (12.5%). In the second group the following CT signs were observed: peritoneal or retroperitoneal fluid (87.5%), bowel wall thickening (50%), mesenteric hematoma (37.5%), sentinel clot (25%), pneumoperitoneum (12.5%), retropneumoperitoneum (12.5%), hazy streaky changes in mesenteric fat (12.5%). DISCUSSION AND CONCLUSIONS: Bowel and mesenteric injuries from blunt trauma are infrequent and difficult to diagnose clinically, as the physical and laboratory findings may be subtle and are often overshadowed by other injuries in patients with multisystem trauma. CT represents a proven modality in the evaluation of bowel and mesenteric injuries: careful inspection and technique are required to detect often subtle findings. In our series of 24 patients with surgically confirmed bowel and mesenteric injuries, the presence of mesenteric hematoma and of peritoneal or retroperitoneal fluid were the more frequent CT signs observed. Radiologists may play a crucial role in the timely diagnosis of these injuries, allowing prompt and appropriate management of these patients. PMID- 11402958 TI - Effects in vivo of iohexol and diatrizoate on human plasma acetyl- and butyryl cholinesterase activity. AB - PURPOSE: The aim of this study was to evaluate the effects of two iodinate contrast agents (CA), iohexol and diatrizoate, on human plasma acetyl-(AC) and butyrylcholinesterase(BC) activity. MATERIAL AND METHODS: Forty-eight patients (24 males and 24 females) scheduled for intravenous pyelography were randomly divided into four groups of 6 males and 6 females each, receiving as CA, respectively: iohexol (Omnipaque, Schering) 0.6 ml/kg body weight (G1); iohexol 1.2 mg/kg (G2); sodium and meglumine diatrizoate 58% (Urografin, Schering) 0.6 ml/kg (G3); sodium and meglumine diatrizoate 58% 1.2 ml/kg (G4). Blood samples were taken before and 5, 10, and 20 min after the injection. Enzymatic activity of AC and BC were measured by spectrophotometry. Plasma concentration of K, Na, Ca, and Mg was measured in all blood samples; blood pressure and plasma pH were measured after each sample collection. Statistical analysis was performed by Student's test. RESULTS: In G1 a reversible decrease of AC (12.9%) and BC (8.2%) plasma activity was observed at 10 min. In G2 a progressive decrease of AC (13.9%) and BC (18.4%) plasma activity was observed with a maximum at 20 min. In G3 a modest reversible decrease of BC plasma activity (5.4%) was observed. In G4 a modest progressive decrease of AC (7.3%) and BC (6.5%) plasma activities was observed. In all cases, AC and BC plasma activities remained within the normal range of values. Plasma concentration of K, Na, Ca, and Mg, as well as pH and systolic and diastolic pressure, did not show any change. No adverse effects was observed in our patients. CONCLUSIONS: Iohexol and diatrizoate induce in vivo a significant decrease of AC and BC plasma activities. The decrease is more pronounced for iohexol, a non ionic CA, which has a lower pharmacotoxicity than diatrizoate and adverse effects rate. No inference can be drawn about the relationship between plasma cholinesterase activity and adverse effects. PMID- 11402959 TI - [Physical-dosimetric characterization of a multi-leaf collimator system for clinical implementation in conformational radiotherapy]. AB - INTRODUCTION: In the present paper we discuss the main dosimetric characteristics of the multileaf collimator (MLC) installed on the Elekta SLi Precise accelerators. To evaluate the effectiveness of the MLC in conformal radiotherapy, beam transmission through leaves and/or diaphragms, leakage between the leaves, central axis depth dose, surface dose, effective penumbra, scalopping effect and field size factors were measured. MATERIALS AND METHODS: The MLC installed on the dual energy (4 and 6 MV) linear accelerator Elekta SLi Precise consists of 40 opposed pairs of 75 mm thick tungsten leaves, set in two raws mounted in place of the upper collimator. Each leaf has a nominal projected width of 10 mm. The maximum field size attainable is 40 x 40 cm2 at 100 cm SAD. Beam transmission through leaves and/or diaphragms and field size factors were measured in RW3 phantom with a ionization chamber, leakage between the leaves and effective penumbra were instead evaluated with radiographic films (X-Omat-V) and a laser scanning photodensitometer. Percentage depth doses were measured in an automatic water phantom. RESULTS: For both energies, approximately 1% of the incident radiation on the multileaf collimator is transmitted through the backup collimator, while the transmission through the different combinations of leaves and collimators is between 0.03 and 0.14%. These values show a good agreement with literature data and are in general lower than the peak values specified by the manufacturer. The peak value of the leakage between the leaves was about 2% for both energies, without significative variation with gantry or collimator angle or distance from the axis. MLC shaped fields show a skin dose less (about 3%) than the one of cerrobend block shaped fields, because of the electronic contamination due to the plexiglass tray of the cerrobend blocks; in both cases, the depth doses are similar, as are flatness and symmetry of irradiation fields. The effective penumbra increases with field dimension, depth and leaves positioning, with a mean value of about 9 mm for both energies. The different beam configurations do not significantly affect the values of the field size factors. CONCLUSIONS: The dosimetric characteristics and the case of use of the Elekta multileaf collimator make its application to conformal radiotherapy convenient and reliable, able to improve the accuracy and the effectiveness of radiation therapy and to develop new kinds of treatments. However, because of the complexity of the MLC, its implementation in radiotherapic practice requires careful dosimetric characterization to evaluate those parameters (transmission, penumbra and output factors) that play a fundamental role in the accuracy of the treatment. PMID- 11402960 TI - [Unusual cerebral ultrasonographic feature in a case of neonatal hypoxia-ischemia distress]. PMID- 11402961 TI - [Flexor sheath hemorrhage of the hand during anticoagulant therapy: CT and MR]. PMID- 11402962 TI - [Evaluation with color-Doppler ultrasonography and spiral CT of a case of adventitial cystic disease of the popliteal artery. Clinical considerations and differential diagnosis]. PMID- 11402963 TI - [Wandering spleen: unusual case of pelvic mass]. PMID- 11402964 TI - [Role of spiral computerized tomography and of ventilatory and perfusion scintigraphy in the diagnosis of pulmonary embolism: method comparison in 2 cases]. PMID- 11402965 TI - [Simple paradigms of obesity]. PMID- 11402966 TI - [Obesity, diet and physical activity]. PMID- 11402967 TI - [Obesity among children--with particular reference to Danish circumstances]. AB - Obesity is one of the most important predictors and causes of such lifestyle diseases as metabolic syndrome, type 2 diabetes, and cardiovascular disease. It is thus imperative to follow the trend longitudinally in the population. Measurement of obesity in children and adolescents is difficult; many studies use the body mass index as a measure, regardless of the fact that it is inappropriate. Skinfolds or the ponderal index should be used, instead. Recent Danish studies suggest that the incidence of childhood and adolescent obesity is increasing and that those who are overweight are even more so today than earlier. Obesity is caused by a number of factors, genetic, social, environmental, and lifestyle, all of which play an important role. One of the main causes of the increase in childhood obesity in Denmark today is the lower level of physical activity than formerly. The prospects for the future are an increase in obesity and incidence of lifestyle diseases with a poorer quality of life and a shorter life time expectancy. PMID- 11402968 TI - [Fatty tissue as a secretory organ. Significance for obesity-related diseases]. AB - Adipose tissue is not only a passive storage organ for excessed energy intake, it is also able to produce and release several substances with local (autocrine) and systemic (endocrine) actions. An up-to-date review of our knowledge in this area is given here. Several of the compounds deriving from adipose tissue have been shown to play a role in obesity-related health complications. The production of cytokines (TNF-alpha, IL-6, IL-8) is implicated in the development of insulin resistance and atherosclerosis. All elements in the renin-angiotensin system are produced in adipose tissue, which is thus related to hypertension. The production of PAI-1 could be related to enhanced thrombogenesis. The release of the compounds described is generally higher from adipocytes in the visceral depot, which could explain the close association between this depot and health complications. PMID- 11402969 TI - [Appetite regulation--leptin and other signal substances]. PMID- 11402970 TI - [Fatty tissue cell metabolism]. PMID- 11402971 TI - [Atherosclerosis and blood lipoproteins]. PMID- 11402972 TI - [What are the characteristics of overweight and obese individuals?]. PMID- 11402973 TI - [Slimming behavior among Danes assessed by telephone interviews in 1992 and 1998]. AB - INTRODUCTION: With the increasing prevalence of overweight and obesity, it is assumed that slimming is on the increase. The aim of the present study was to elucidate the prevalence of slimming attempts, the methods used, and whether the weight loss was maintained. METHODS: Two independent telephone interviews were conducted in 1992 and in 1998. About 1200 adults selected at random were entered in each survey in a balanced design, which ensured an equal distribution of age, gender, and geographical regions in Denmark. RESULTS: The prevalence of obesity increased from 10% to 13% between 1992 and 1998. The proportion of subjects who had been slimming at least once in their lives increased from 45% to 47%, with a distribution of 25% in the underweight, 38% in those of normal weight, 54% of the overweight, and 76% of the obese (p < 0.0001). The use of anti-obesity medicine on prescription and support by a primary care physician or dietician increased with increasing body mass index. Subjects who chose to change their diet or increase their physical activity were more successful in achieving and maintaining a weight loss than those who did not. Changes in diet rose significantly from 1992 to 1998, whereas the use of prescription medicine and over-the-counter protein formulas and supplements decreased. The use of prescription medicine by subjects of normal weight decreased from 11% to 2%. CONCLUSION: About half of all adult Danes have been on a slimming diet. Those who choose to change their diet or increase physical activity are more successful in achieving a weight loss and maintaining it. Notably, one in four underweight subjects have been on a slimming diet, and one in four obese subjects have never dieted, particularly obese males. The use of prescription slimming compounds by individuals of normal weight seems to be a decreasing problem. PMID- 11402974 TI - [The effect of sibutramine for the maintenance of weight loss. A randomized controlled clinical trial]. AB - INTRODUCTION: Sibutramine is a tertiary amine that has been shown to induce dose dependent weight loss and enhance the effects of a low-calorie diet for up to a year. We did a randomised, double-blind trial to assess the usefulness of sibutramine in maintaining substantial weight loss over 18 months. METHODS: Eight European centres recruited 605 obese patients (BMI 30-45 kg/m2) for a 6-month period of weight loss with sibutramine (10 mg/day) in combination with an individualised dietary deficit programme of 600 kcal/day based on the measured resting rates of energy expenditure. Of these 605, 467 (77%) patients with more than a 5% weight loss were then randomly assigned to 10 mg/day sibutramine (n = 352) or placebo (n = 115) for a further 18 months. Sibutramine was increased up to 20 mg/day if a weight regain occurred. RESULTS: One hundred and forty-eight (42%) subjects in the sibutramine group and 58 (50%) in the placebo group dropped out. Of the 204 subjects receiving sibutramine who completed the trial, 89 (43%) maintained 80% or more of their original weight loss, compared with nine (16%) of the 57 subjects in the placebo group (odds ratio 4.64, p < 0.001). Substantial decreases were seen over the first six months in triglycerides, VDL cholesterol, insulin, C peptide, and uric acid; changes, which were sustained in the sibutramine group, but not in the placebo group. Concentrations of HDL cholesterol rose substantially in the second year: overall increases were 20.7% (sibutramine) and 11.7% (placebo, p < 0.001). Twenty (3%) patients were withdrawn because of raised blood pressure; in the sibutramine group, from baseline to two years systolic blood pressure rose by 0.1 mmHg (SD 12.9), diastolic blood pressure by 2.3 mmHg (9.4), and pulse rate by 4.1 beats/min (11.9). CONCLUSION: This individualised management programme achieved a weight loss in 77% of obese patients and a sustained weight loss in most patients continuing therapy for two years. Changes in the concentrations of HDL cholesterol, VDL cholesterol, and triglyceride, but not in the LDL cholesterol, exceed those expected either from a weight loss alone or when induced by other selective therapies for low concentrations of HDL cholesterol relating to coronary heart disease. PMID- 11402975 TI - [The significance of diet and physical activity for the development of obesity in Denmark from 1985 to 1995]. AB - MATERIAL AND METHODS: Trends in the prevalence of overweight and obesity in Denmark were compared to changes in diet and physical activity by using data from the nationwide dietary surveys of 1985 and 1995 comprising 2,000 adults, aged 15 80 years. RESULTS: The prevalence of overweight and obesity has increased by more than 20% in men (1985: 35.7% vs 1995: 44.1%) and women (1985: 22.3% vs 1995: 29.4%). The prevalence of obesity rose especially in men from 1985 (5.0%) to 1995 (8.8%), less dramatically in women (1985: 5.3% vs 1995: 6.6%). A decrease of 20% and 5% in energy intake has been registered, respectively in men and women. At the same time, sedentary work increased considerably from 1995 (19.5%) to 1985 (38.2%), whereas leisure time physical activity was higher than before. DISCUSSION: The increasing prevalence of overweight and obesity is probably explained by a decrease in the total energy expenditure, owing to the fact that sedentary work has doubled. Hence, a rise in the level of leisure time physical activity did not compensate for the lower expenditure of energy during working hours. PMID- 11402976 TI - [Beer, wine, spirits and mortality. Results from a prospective population study]. AB - INTRODUCTION: The aim of the present population-based cohort study was to examine the association between alcohol intake and mortality from all causes, coronary heart disease, and cancer. METHODS: A prospective population study with baseline assessment of beer, wine and spirit consumption, smoking habits, educational level, physical activity, and body mass index in a total of 257,859 person-years follow-up on mortality. RESULTS: A total of 4833 participants died, 1075 of these from coronary heart disease and 1552 of cancer. Compared with non-drinkers, light drinkers, who avoided wine, had a relative risk of death from all causes of 0.90 (0.82-0.99) and those who drank wine had a relative risk of 0.66 (0.55-0.77). Heavy drinkers, who avoided wine, were at higher risk of death from all causes than were heavy drinkers, who included wine in their alcohol consumption. Wine drinkers had a significantly lower mortality from both coronary heart disease and cancer than had non-wine drinkers (p = 0.007 and p = 0.004, respectively). CONCLUSION: A moderate consumption of wine may have a beneficial effect on all causes of mortality, which is additive to that of alcohol. This effect may be attributable to a reduction in death from both coronary heart disease and cancer. PMID- 11402977 TI - [Resistin, another brand-new hormone]. PMID- 11402978 TI - [Bartonella bacterium is suspected as the cause of sudden death among Swedish cross-country runners]. PMID- 11402979 TI - [When radiation waves are more hazardous than microwaves]. PMID- 11402980 TI - [Modern treatment of psychoses?]. PMID- 11402981 TI - [There are still children who are abused--in spite of new laws and increased awareness concerning children's rights]. PMID- 11402982 TI - [Munchausen by proxy syndrome. When a parent who seems to engaged falsifies symptoms of his child]. PMID- 11402983 TI - [Increasing number of child abuse cases in Sweden--in accordance with reality?]. AB - The number of police reports on child assault shows an increasing trend during the last two decades in Sweden. The purpose of this article is to present possible explanations. Increased awareness of child abuse, legislative reforms, changes in attitudes toward corporal punishment and violence in general, and changed routines within schools can explain the increasing trend in police reporting. An actual increase in the rate of child abuse is possible but less likely. Reports of increasingly violent behavior among young people must be taken seriously. PMID- 11402984 TI - [Munchausen syndrome by proxy. Child abuse by falsification of symptoms]. AB - Munchausen syndrome by proxy (MSbP) is a potentially lethal form of child abuse. Mortality rates around 10 percent have been reported. A knowledge of the characteristics of MSbP is needed in order to reveal the diagnosis and to handle the management correctly. Swedish experience is in accordance with international reports. PMID- 11402985 TI - [Antisocial behavior of adolescents and home environment--need for a multidisciplinary model]. AB - In Swedish child and adolescent psychiatry there is a more than 60-year long tradition of using longitudinal methods in research on juvenile delinquency. Since the 1940's, results have been presented using either prospective or retrospective longitudinal designs for this purpose. Starting from genetics as scientific paradigm, new approaches including neuropsychiatry and social psychiatry showed the need for a multidisciplinary view at the border between medicine and behavioral sciences. Both Swedish and international research in the area has clearly demonstrated that factors relating to gender, maturation, resilience vs. vulnerability, the mental health of the parents, the social network and the organization of the school are of importance when trying to understand, prevent and treat juvenile delinquents. The challenge presented to today's and future researchers is to understand how the complexity of the modern western society will impact already established knowledge. PMID- 11402986 TI - [Experiences from the county of Norrbotten: screening of hearing at the age of four crucial for detecting mild hearing impairment]. AB - Since the eighties the formerly well established hearing screening performed at the age of four has been debated, partly for economic reasons. One quarter of Sweden's counties have since then abolished it. But how many children do we detect by this screening? What significance does it have for a child that those around it are aware of a mild hearing impairment? In Norrbotten, in all, we found 123 children born 1989-1992 with a hearing impairment. The prevalence was 7.9 per thousand. 30 percent of the children were detected at the 4-years hearing screening, and of these 0.9/1,000 had a hearing impairment that was bilateral, 1.0/1,000 unilateral and 0.5/1,000 impairment only in the high frequencies. Also children with mild bilateral or unilateral hearing impairment can experience difficulties in school and among friends, especially in noisy surroundings, and benefit by knowing early about their hearing impairment. PMID- 11402987 TI - [Measurements in child health services. Also experienced nurses obtain different results]. AB - Growth reflects general health but also the nutritional and environmental health of a child. Disordered growth may be an early manifestation of disease or deficiency. In Sweden, the newly updated growth charts are highly valued at the child health services, yet measurements of height, weight and head circumference of four infants showed that even experienced nurses achieved noticeably large differences in their individual results. In several cases the plotted data also deviated concerning the exact age of the child or the registered value. Awareness of methodological problems should lead to more accurate advice and referral. PMID- 11402988 TI - [A study at a health center. "Guide" in the Fass can result in overprescription of penicillin to children]. PMID- 11402989 TI - [Hepatitis B outbreak in a day care center affected several families. It could have been prevented by vaccination of all children]. AB - An outbreak of hepatitis B originating in a family day nursery affected several children with Somali background. The transmission chain was confirmed by sequence analysis of the S gene. In Africa hepatitis B is often spread horizontally among children of pre-school age, a pattern of transmission that was retained in this outbreak. To limit the outbreak 126 children in the nursery and 50 members of staff had to be vaccinated. The total cost for this intervention was estimated to about 300,000 SEK. Considering the great number of immigrants in Sweden from areas highly endemic for hepatitis B the inclusion of vaccination in the general child immunisation program seems to be the most cost effective measure for long term prevention not only of hepatitis B transmission among children but also of venereal spread in early adulthood. PMID- 11402990 TI - [A case report. Drugs can be administered via colostomy but the scientific support is negligible]. PMID- 11402991 TI - [50th anniversary of the import of physicians from Austria to Sweden]. PMID- 11402992 TI - [A study of the utilization of radiology at health centers in Jarfalla. The average cost of radiology was 111 crowns per capita]. PMID- 11402993 TI - [Bologna--a city with close ties to medical history]. PMID- 11402994 TI - [What happened with the problem of hearing impaired persons?]. PMID- 11402995 TI - [Too optimistic view in mass media on statin treatment in acute myocardial infarction]. PMID- 11402996 TI - [The debate on circumcision--some interpretations take precedence]. PMID- 11402997 TI - [Safe care is a right also in connection with male circumcision]. PMID- 11402998 TI - [Circumcision or genital mutilation--more than a terminological dispute]. PMID- 11402999 TI - [A wish to change the tradition]. PMID- 11403001 TI - American Diabetes Association Clinical Practice Recommendations 2001. PMID- 11403000 TI - [Painful procedures performed in children who were not informed]. PMID- 11403002 TI - Web alert. PMID- 11403003 TI - Web alert. PMID- 11403004 TI - Web alert. PMID- 11403005 TI - Seminar on Health Research Ethics in Africa. 1-4 November 1999, Arusha, Tanzania. PMID- 11403006 TI - The global drug gap: policy implications for microbicides. PMID- 11403007 TI - Chemokine receptors and HIV entry. PMID- 11403008 TI - The development of novel vaginal microbicides: from the bench to the clinic. PMID- 11403009 TI - The ethics of testing microbicides: are we protecting the volunteers or the investigators? PMID- 11403010 TI - Microbicides 2000 conference. Panel discussion: streamlining the process and regulatory issues. Microbicide research-streamlining the process. PMID- 11403011 TI - 91st annual meeting of the American Association for Cancer Research. PMID- 11403012 TI - [Intraoperative hypothermia]. PMID- 11403013 TI - [23rd International Congress of the Association of French-speaking Dermatologists. 22-26 February 2001, Cayenne. Abstracts]. PMID- 11403014 TI - European Congress of Radiology. March 2-6, 2001. Vienna, Austria. Abstracts. PMID- 11403015 TI - Biochemical Society 671st meeting. University of Leeds. Abstracts. PMID- 11403016 TI - The 41st Annual Meeting of the Japanese Respiratory Society. April 4-6, 2001. Tokyo, Japan. Abstracts. PMID- 11403018 TI - 74th Annual Meeting of the Japanese Pharmacological Society. Yokohama, Japan, March 21-23, 2001. Abstracts. PMID- 11403017 TI - 33rd Annual Meeting of the American Burn Association. April 18-21, 2001. Boston, Massachusetts, USA. Abstracts. PMID- 11403019 TI - 1st International Workshop on Adverse Drug Reactions and Lipodystrophy in HIV. 26 28 June 1999, San Diego, California, USA. Abstracts. PMID- 11403020 TI - [Diabetes meeting. Montpellier, France, 27-31 March 2001. Abstracts]. PMID- 11403021 TI - Integrated bioinformatics- high-throughput interpretation of pathways and biology. PMID- 11403022 TI - Challenges in bioinformatics: infrastructure, models and analytics. PMID- 11403023 TI - William Farr's influence on Florence Nightingale. PMID- 11403024 TI - 67th Annual Meeting of the German Society of Cardiology--Heart and Circulatory Research. 19-21 April 2001, Mannheim. Abstracts. PMID- 11403025 TI - British Pharmacological Society Meeting. 18-21 December 2000, University of Birmingham. Abstracts. PMID- 11403027 TI - 29th European Symposium on Clinical Pharmacy: New Technologies, Pharmacists and Patients. 11-14 October 2000, Basel, Switzerland. Abstracts. PMID- 11403026 TI - Dental schools or institutes: upon whom will we rely? PMID- 11403028 TI - Progress in practice: United Kingdom Clinical Pharmacy Association autumn symposium. Blackpool, 24-26 November 2000. Abstracts. PMID- 11403029 TI - American Society of Hypertension 16th annual meeting. May 15-19, 2001. San Francisco, California, USA. Abstracts. PMID- 11403030 TI - Annual Clinical Genetics Meeting. March 9-12, 2000. Palm Springs, California, USA. Abstracts. PMID- 11403031 TI - [Francophile meeting of digestive pathology. Paris, France, 24-28 March 2001. Abstracts]. PMID- 11403032 TI - The 7th ESACP (European Society for Analytical Cellular Pathology) Congress. Caen, France. April 1-5, 2001. Abstracts. PMID- 11403033 TI - 2000 Andrea Prader Prize. Summary of Andrea Prader Lecture 2000. PMID- 11403034 TI - Recommendations for the registration of drugs used in the treatment of osteoarthritis: an update on biochemical markers. PMID- 11403035 TI - CFC transition. PMID- 11403036 TI - CFC transition. PMID- 11403037 TI - Time to consign cromoglycate to history? PMID- 11403038 TI - 141st Meeting of the Acoustical Society of America. Chicago, Illinois, USA. 4-8 June 2001. Abstracts. PMID- 11403039 TI - SHOX point mutations in dyschondrosteosis. PMID- 11403040 TI - Phenotypic heterogeneity of CYP1B1: mutations in a patient with Peters' anomaly. PMID- 11403041 TI - Role of TP53 P72R polymorphism in human papillomavirus associated premalignant laryngeal neoplasm. PMID- 11403042 TI - Predisposing chromosome for spinocerebellar ataxia type 6 (SCA6) in Japanese. PMID- 11403043 TI - Prenatal testing for Huntington's disease: experience within the UK 1994-1998. PMID- 11403044 TI - Pregnancy outcome and long term prognosis in 868 children born after second trimester amniocentesis for maternal serum positive triple test screening and normal prenatal karyotype. PMID- 11403045 TI - A syndrome of overgrowth and acromegaloidism with normal growth hormone secretion is associated with chromosome 11 pericentric inversion. PMID- 11403046 TI - Maternally inherited duplication of the possible imprinted 14q31 region. PMID- 11403047 TI - A novel missense mutation in the GTPase activating protein homology region of TSC2 in two large families with tuberous sclerosis complex. PMID- 11403048 TI - Interstitial deletion of 3p22.2-p24.2: the first reported case. PMID- 11403049 TI - [The latest information of the schools of public health in USA: with reference to a proposed school of public health in Japan at the 21st century]. PMID- 11403050 TI - Post-traumatic stress disorder. Doctors should relieve suffering, not debate its existence. PMID- 11403051 TI - Post-traumatic stress disorder. Disorder takes away human dignity and character. PMID- 11403052 TI - Post-traumatic stress disorder. Accountable methods of validation are needed. PMID- 11403053 TI - Post-traumatic stress disorder. Logic is flawed. PMID- 11403054 TI - Post-traumatic stress disorder. Social usefulness of any diagnosis needs consideration. PMID- 11403055 TI - Post-traumatic stress disorder. Diagnostic label is missing. PMID- 11403056 TI - Post-traumatic stress disorder. Baby should not be thrown out with bath water. PMID- 11403057 TI - Post-traumatic stress disorder. Questions about current status of psychiatric classification systems arise. PMID- 11403058 TI - How policy informs the evidence. Comprehensive evidence is needed in decision making. PMID- 11403059 TI - How policy informs the evidence. Lessons have also been learnt in disciplines outside medicine. PMID- 11403060 TI - Riluzole for motor neurone disease. More trials are needed. PMID- 11403061 TI - Riluzole for motor neurone disease. Large, more economical trial is needed. PMID- 11403062 TI - Riluzole for motor neurone disease. Ambiguous diagnostic criteria mean that wide variations in elgibility may still persist. PMID- 11403063 TI - French cardiovascular mortality did not increase during 1996 European football championship. PMID- 11403064 TI - Athlete's foot and fungally infected toenails. Authors should use familiar drug names. PMID- 11403065 TI - Medical students see that academic misconduct is common. PMID- 11403066 TI - Single use disposable equipment is not being used everywhere. PMID- 11403067 TI - Time required for partial pressure of arterial oxygen equilibration during mechanical ventilation after a step change in fractional inspired oxygen concentration. AB - OBJECTIVE: To determine the time required for the partial pressure of arterial oxygen (PaO2) to reach equilibrium after a 0.20 increment or decrement in fractional inspired oxygen concentration (FIO2) during mechanical ventilation. SETTING: A multi-disciplinary ICU in a university hospital. PATIENTS AND METHODS: Twenty-five adult, non-COPD patients with stable blood gas values (PaO2/FIO2 > or = 180 on the day of the study) on pressure-controlled ventilation (PCV). Following a baseline PaO2 (PaO2b) measurement at FIO2 = 0.35, the FIO2 was increased to 0.55 for 30 min and then decreased to 0.35 without any other change in ventilatory parameters. Sequential blood gas measurements were performed at 3, 5, 7, 9, 11, 15, 20, 25 and 30 min in both periods. The PaO2 values measured at the 30th min after a step change in FIO2 (FIO2 = 0.55, PaO2[55] and FIO2 = 0.35, PaO2[35]) were accepted as representative of the equilibrium values for PaO2. Each patient's rise and fall in PaO2 over time, PaO2(t), were fitted to the following respective exponential equations: PaO2b + (PaO2[55]-PaO2b)(1-e-kt) and PaO2[55] + (PaO2[35]-PaO2[55])(e-kt) where "t" refers to time, PaO2[55] and PaO2[35] are the final PaO2 values obtained at a new FIO2 of 0.55 and 0.35, after a 0.20 increment and decrement in FIO2, respectively. Time constant "k" was determined by a non-linear fitting curve and 90% oxygenation times were defined as the time required to reach 90% of the final equilibrated PaO2 calculated by using the non-linear fitting curves. RESULTS: Time constant values for the rise and fall periods were 1.01 +/- 0.71 min-1, 0.69 +/- 0.42 min-1, respectively, and 90% oxygenation times for rises and falls in PaO2 periods were 4.2 +/- 4.1 min-1 and 5.5 +/- 4.8 min-1, respectively. There was no significant difference between the rise and fall periods for the two parameters (p > 0.05). CONCLUSION: We conclude that in stable patients ventilated with PCV, after a step change in FIO2 of 0.20, 5-10 min will be adequate for obtaining a blood gas sample to measure a PaO2 that will be representative of the equilibrium PaO2 value. PMID- 11403069 TI - [Painful joint diseases]. PMID- 11403068 TI - Hirudin versus heparin for anticoagulation in continuous renal replacement therapy. AB - OBJECTIVE: To compare the efficacy and safety of hirudin and heparin for anticoagulation during continuous renal replacement therapy (CRRT) in critically ill patients. DESIGN: Prospective, randomized controlled pilot study. SETTING: Single centre; interdisciplinary intensive care unit at a university hospital. PATIENTS: Seventeen patients receiving CRRT. INTERVENTIONS: Patients were randomly allocated to two groups. Heparin group (nine patients): continuous administration of 250 IU/h heparin; dose was adjusted in 125 IU/h steps with a targeted activated clotting time (ACT) of 180-210 s. Hirudin group (eight patients): continuous infusion of 10 micrograms/kg/h hirudin, dose was adjusted in 2 micrograms/kg/h steps with a targeted ecarin clotting time (ECT) of 80-100 s. Observation time was 96 h. MEASUREMENTS AND MAIN RESULTS: Measured filter run patency and haemofiltration efficacy did not significantly differ between the two groups. Three bleeding complications were observed in the hirudin group, none in the heparin group (P < 0.01). At the onset of bleeding, which occurred 60 or more hours after the start of therapy, only one patient was still under continuous hirudin administration but levels were either in therapeutic range or below. CONCLUSIONS: Hirudin can be used efficiently for anticoagulation in CRRT. Late bleeding complications may have been caused by possible hirudin accumulation, but this was not evident from hirudin plasma and ECT levels. Since bleeding complications were observed only in the presence of documented coagulation disorders, not only adequate drug monitoring but also the plasmatic and cellular coagulation status of the patient should be taken into consideration for adjusting hirudin dosage. PMID- 11403071 TI - Leprosy--forgotten but not eradicated. PMID- 11403072 TI - Methods for studying biofilms produced by Staphylococcus epidermidis. PMID- 11403073 TI - Age of puberty among girls and the diagnosis of precocious puberty. PMID- 11403074 TI - Adverse sedation events in pediatrics. PMID- 11403075 TI - The two Lyme camps. PMID- 11403076 TI - Privacy protection. PMID- 11403077 TI - Benefits of asthma education programs. PMID- 11403078 TI - Increasing identification of psychosocial problems. PMID- 11403079 TI - Esophageal coins. PMID- 11403080 TI - Use of psychoactive medication during pregnancy and possible effects on the fetus and newborn. PMID- 11403081 TI - Warning: are you aware of "holding therapy"? PMID- 11403082 TI - A facile and effective synthesis of alpha-(1-->6)-linked mannose di-, tri-, tetra , hexa-, octa-, and dodecasaccharides, and beta-(1-->6)-linked glucose di-, tri-, tetra-, hexa-, and octasaccharides using sugar trichloroacetimidates as the donors and unprotected or partially protected glycosides as the acceptors. AB - Reaction of 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl trichloroimidate with allyl alpha-D-mannopyranoside in the presence of TMSOTf selectively gave allyl 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl-(1-->6)-alpha-D-mannopyranoside through an orthoester intermediate. Benzoylation of 3, followed by deallylation, and then trichloroimidation afforded the disaccharide donor 2,3,4,6-tetra-O acetyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl trichloroimidate, while benzoylation of 3 followed by selective removal of acetyl groups yielded the disaccharide acceptor allyl alpha-D-mannopyranosyl-(1-->6) 2,3,4-tri-O-benzoyl-alpha-D-mannopyranoside. Coupling of 5 with 6 gave the tetrasaccharide allyl 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4 tri-O-benzoyl-alpha-D-mannopyranosyl-(1-->6)-alpha-D-mannopyranosyl-(1-->6)-2,3,4 tri-O-benzoyl-alpha-D-mannopyranoside, which were converted into the tetrasaccharide donor 2,3,4,6-tetra-O-acetyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4 tri-O-benzoyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl trichloroimdate and the tetrasaccharide acceptor allyl alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O benzoyl-alpha-D-mannopyranosyl-(1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranosyl (1-->6)-2,3,4-tri-O-benzoyl-alpha-D-mannopyranoside, respectively, by the same strategies as used for conversion of 3 into 5 and 6. Condensation of 5 with 13 gave the hexasaccharide 14, while condensation of 12 with 13 gave the octasaccharide 17. Dodecasaccharide 21 was obtained by the coupling of 12 with the octasaccharide acceptor 20. Similar strategies were used for the syntheses of beta-(1-->6)-linked glucose di-, tri-, tetra-, hexa-, and octamers. Deprotection of the oligosaccharides in ammonia-saturated methanol yielded the free alpha-(1- >6)-linked mannosyl and beta-(1-->6)-linked glucosyl oligomers. PMID- 11403083 TI - Synthesis of C-mannopyranosylphloroacetophenone derivatives and their anomerization. AB - The reaction of 2,3,4-tri-O-benzyl-alpha-L-rhamnopyranosyl fluoride (6-deoxy 2,3,4-tri-O-benzyl-alpha-L-mannopyranosyl fluoride) with 2,4 dibenzylphloroacetophenone, in the presence of boron trifluoride.diethyl etherate, afforded both the 3-C-alpha-L- and the 3-C-beta-L rhamnopyranosylphloroacetophenone derivatives. The 3-C-alpha-L-rhamnoside was produced as a major product, while the 3-C-beta-L-rhamnoside was produced as a minor product via anomerization of the 3-C-alpha-L-rhamnoside. Alternatively, the reaction of 2,3,4,6-tetra-O-benzyl-alpha-D-mannopyranosyl fluoride with 2,4 dibenzylphloroacetophenone afforded both the 3-C-alpha-D- and the 3-C-beta-D mannnopyranosylphloroacetophenone derivatives under identical conditions. The 3-C beta-D-mannoside was produced as a major product via anomerization of the 3-C alpha-D-mannoside during the reaction. These differences in composition result apparently from the magnitude of the 1,3-diaxial interactions between the C-3 and C-5 positions in these sugar moieties. PMID- 11403084 TI - Structural elucidation of a new arabinogalactan from the leaves of Nerium indicum. AB - A polysaccharide fraction, NIB-2, was obtained from the 3% aqueous sodium carbonate extract of Nerium indicum leaves using anion-exchange chromatography and gel-permeation chromatography. It was found to be composed of rhamnose, arabinose, galactose, in the ratios of 1.0:10.4:4.4, along with 4% of galacturonic acid. The results of methylation analysis, periodate oxidation, partial acid hydrolysis, pectinase treatment, and 13C and 1H NMR spectroscopy indicate that it is mainly an arabinogalactan having a backbone of 1,6-linked beta-Galp, with branches at O-3, consisting of terminal, 1,5-, and 1,3,5-linked arabinofuranosyl residues, and a small proportion of galactosyl residues at the termini. Rhamnose and galacturonic acid arose from a contaminating rhamnogalacturonan. PMID- 11403085 TI - Synthesis of a carbocyclic sialic acid analogue for the inhibition of influenza virus neuraminidase. AB - The influenza virus neuraminidase (NA) is essential for viral infection and offers a potential target for antiviral drug development. We prepared a carbocyclic sialic acid analogue, potentially able to inhibit NA. Its structure is an analogue of the transition-state of the reaction catalysed by NA. As starting material, quinic acid was selected owing to its ready availability and its stereochemical feature suitable for the target structure. The quinic acid was first converted in the shikimic acid; then two of the three hydroxyl functions of this product were selectively functionalised to obtain the target molecule (3R,4S,5R)-4-acetamido-3-guanidino-5-hydroxycyclohex-1-ene-1-carboxylic acid. PMID- 11403086 TI - Transglucosidation of methyl and ethyl D-glucofuranosides by alcoholysis. AB - The acid catalyzed ethanolysis of methyl 5-O-methyl-alpha- and -beta-D glucofuranoside and the analogous methanolysis of ethyl 5-O-methyl-alpha- and beta-D-glucofuranoside have been investigated. For all four reactions, the primarily formed transglycosylation product was a single glycoside that had the opposite anomeric configuration to the starting material. This strongly indicates that a D-glucose methyl ethyl acetal is first formed and is then ring closed by a nucleophilic attack by HO-4, giving either the starting material or a transglycosylation product with the opposite anomeric configuration. Low percentages of the methyl ethyl acetals and of dimethyl acetals were also observed in the reaction product during the methanolysis reactions. PMID- 11403087 TI - Synthesis of linear-type chondroitin clusters having a C8 spacer between disaccharide moieties and enzymatic transfer of D-glucuronic acid to the artificial glycans. AB - Newly designed linear-type glycoclusters were synthesized which involve a chondroitin repeating disaccharide ligand and a hydrophobic octyl ether spacer. The spacer mimics the corresponding disaccharide unit. Repeating elongation of the pseudo-tetrasaccharide that was derived from the common cluster unit [-->8) octyl-(1-->3)-beta-D-Gal-NAc-(1-->4)-beta-D-GlcA-(1-->] allowed the syntheses of up to the pseudo-decasaccharide analog of chondroitin. An enzymatic D-GlcA transfer at the non-reducing end of the synthesized artificial glycans by GlcATase II was observed. PMID- 11403088 TI - Xylopyranoside-based agonists of D-myo-inositol 1,4,5-trisphosphate receptors: synthesis and effect of stereochemistry on biological activity. AB - The synthesis of a series of tetrahydrofuranyl alpha- and beta-xylopyranoside trisphosphates, designed by excision of three motifs of adenophostin A is reported. The synthetic route features improved preparations of allyl alpha-D xylopyranoside and its 2-O-benzyl ether, and gives access to four diastereoisomeric trisphosphates, which show a range of abilities to mobilise Ca2+ from the intracellular stores of hepatocytes. A comparison of the potencies of the four trisphosphates provides useful information relating to the effects of stereochemical variation on the recognition of carbohydrate-based trisphosphates by D-myo-inositol 1,4,5-trisphosphate receptors. 1-O-[(3'S,4'R)-3 hydroxytetrahydrofuran-4-yl] alpha-D-xylopyranoside 3,4,3'-trisphosphate (8) is the most active member of the series with a potency close to Ins(1,4,5)P3; a beta linked analogue, 1-O-[(3'R,4'S)-3-hydroxytetrahydrofuran-4-yl] beta-D xylopyranoside 3,4,3'-trisphosphate, is ca. 20-fold weaker than Ins(1,4,5)P3, and the other compounds are much less active. While no compound attained a potency close to that of adenophostin A, we believe that 8 represents the minimal structure for potent Ca2+-releasing activity in this type of carbohydrate-based analogue. PMID- 11403089 TI - Structural and immunological studies of a major polysaccharide from spores of Ganoderma lucidum (Fr.) Karst. AB - A polysaccharide isolated from spores of the fungus, Ganoderma lucidum, was found to be a complex glucan. On the basis of compositional and methylation analyses, periodate oxidation, Smith degradation, 1D and 2D NMR, and ESIMS experiments of the native polysaccharide and its degraded products, the polysaccharide was shown to have a backbone of beta-(1-->3)-linked D-glucopyranosyl residues, with branches of mono-, di- and oligosaccharide side chains substituting at the C-6 of the glucosyl residues in the main chain. Conformational analysis in aqueous solution and immunological activities of the native and degraded glucans were also investigated. The results suggested that the degree of substitution on the main chain and the length of side chains may be very important factors in determining the conformation and the biological activities of beta-(1-->3)-linked glucans. PMID- 11403090 TI - Anticoagulant properties of dextranmethylcarboxylate benzylamide sulfate (DMCBSu); a new generation of bioactive functionalized dextran. AB - Dextranmethylcarboxylate benzylamide sulfate (DMCBSu), a functionalized dextran, exhibits anticoagulant properties. Its synthesis involves three steps: a carboxymethylation with monochloroacetic acid in alkaline water-iso-propanol, a benzylamidification of some of the methylcarboxylate groups with benzylamine in the presence of a water soluble carbodiimide and a partial sulfation of the remaining hydroxyl groups with SO3-pyridine in dimethylformamide. This procedure yields reproducibly DMCBSu with degrees of substitution in methylcarboxylate (MC), benzylamide (B) and sulfate (Su) groups, respectively, up to 1.61, 0.35 and 1.5, each obtained in one step. For a degree of substitution of methylcarboxylate ca. 1, the presence of sulfate groups is absolutely necessary to confer anticoagulant activities to the samples. In addition, the anticoagulant ability is higher for derivatives bearing benzylamide groups. The anticoagulant ability of DMCBSu increases with the degree of sulfation, reaching 20% of heparin activity for a degree of substitution of Su groups about 1.3. PMID- 11403091 TI - Structure of the complex of beta-cyclodextrin with beta-naphthyloxyacetic acid in the solid state and in aqueous solution. AB - The structure of the complex of beta-cyclodextrin (cyclomaltoheptaose) with beta naphthyloxyacetic acid was studied in solid state by X-ray diffraction and in aqueous solution by 1H NMR spectroscopy. The complex crystallizes in the channel mode, space group C2, with a stoichiometry of 2:1; two beta-cyclodextrin molecules related by a twofold crystal axis form dimers, in the cavity of which one guest molecule is found on average. The above stoichiometry indicates one guest per beta-CD dimer statistically oriented over two positions or two guest molecules in pi-pi interactions in half of the beta-CD dimers and the rest of the beta-CD dimers empty. In addition, occupancy of 0.5 for the guest per every beta CD dimer is in accord with the occupancy of the two disordered primary hydroxyls. These two hydroxyl groups, to which the carboxylic oxygen atoms of the guest are hydrogen bonded, point towards the interior of the beta-CD cavity. In aqueous solution, the 1H NMR spectroscopic study indicated that there is a mixture of complexes with host-guest stoichiometries both 1:1 and 2:1. PMID- 11403092 TI - Sugar-based anionic surfactants: synthesis and micelle formation of sodium methyl 2-acylamido-2-deoxy-6-O-sulfo-D-glucopyranosides. AB - The following sequence of reactions has been employed to synthesize the title anionic surfactants: [chemical reaction: see text] where R=C7H15; C11H23; and C15H31, respectively, and Py refers to pyridine. Aggregation of the surfactants synthesized (predominantly alpha anomers) in water was studied at 40 degrees C by conductivity measurements. Increasing the chain length of R decreases the critical micelle concentration (CMC) and the degree of counter-ion dissociation. The dependence of the Gibbs free energy of micellization and CMC on the length of R is similar to other ionic surfactants, but the head-group, i.e., the sulfated sugar moiety is less hydrophilic than the structurally related group -(OCH2-CH2)2 OSO3-Na+, most probably because of intermolecular H-bonding in the micellar pseudo-phase PMID- 11403093 TI - A distal radius lesion in a 21-year-old-woman. PMID- 11403094 TI - Physiological roles of corticotropin-releasing hormone receptor type 2. AB - Recent investigations of the physiological roles of CRH-R2 are reviewed and summarized in Fig. 5. VMH CRH-R2 is more important than CRH-R1 in mediating anorexic effect of CRH or urocortin (UCN) and stress-induced reduction of food intake. CRH-R2 mediates a central anxiolytic response, opposing the anxiogenic effect of CRH mediated by CRH-R1. Hippocampal CRH-R1 mediates stress-induced enhancement of learning, while CRH-R2 in the lateral intermediate septum may act to impair learning. CRH-R1 mediates CRH-induced blood pressure elevation, while peripheral CRH-R2 mediates the hypotensive effect of systemically administered UCN and CRH. It is likely that CRH-R2 does not play an important role in hypothalamic-pituitary adrenal axis regulation, though it has been reported that CRH-R2-deficient mice showed hyperresponse of ACTH and corticosterone. Peripheral CRH-R2 mediates UCN-induced mast cell degranulation, vascular permeability, and abdominal surgery-induced gastric stasis. These recent investigations have revealed that the existence of two CRH receptors, which mediate some opposite effects, provides the CRH and UCN systems a high flexibility and dynamic role in the adaptation of the body to environmental challenge. PMID- 11403095 TI - A case of aldosterone-producing adrenocortical adenoma associated with preclinical Cushing's syndrome and hypersecretion of parathyroid hormone. AB - A rare case of aldosterone-producing adrenocortical adenoma with preclinical Cushing's syndrome and hypersecretion of parathyroid hormone (PTH) is described. A 64-year-old male patient had a history of hypertension for two decades and hypokalemia for 4 years. He suffered from left hemiparesis and aphasia due to cerebral hemorrhage, but his appearance was not Cushingoid. His plasma renin activity was below the normal range, while plasma aldosterone concentration was high. They did not respond to furosemide-upright test. His plasma cortisol level in the morning was at the upper limit of the normal range, but it did not show a diurnal rhythm nor was it suppressed by 1 mg and 8 mg of dexamethasone. Computed tomography showed a low density tumor in the right adrenal gland. An adrenal scintigram under dexamethasone treatment revealed an uptake of the tracer on the right side, and plasma aldosterone and cortisol concentrations in the adrenal vein were higher on the right side than on the opposite. The diagnosis of right aldosterone-producing adrenal adenoma with an autonomous production of cortisol was confirmed by right adrenalectomy. Histological findings showed an adenoma consisting mostly of clear cells, but that the nests of compact cells were scattered. Analysis of an extract from the adenoma revealed that the adenoma contained an excess amount of aldosterone and that the cortisol/corticosterone ratio was higher than that of aldosterone-producing adenoma. Both serum calcium and PTH levels remained high one year after adrenalectomy. Ultrasonography revealed the swelling of a parathyroid gland on the left side, indicating the coexistence of an autonomous hyperparathyroidism. PMID- 11403096 TI - Inverse correlation between the changes of lumbar bone mineral density and serum undercarboxylated osteocalcin after vitamin K2 (menatetrenone) treatment in children treated with glucocorticoid and alfacalcidol. AB - We have reported that alfacalcidol plus menatetrenone, a vitamin K2 with four isoprene units (menaquinone-4), treatment is useful for improving bone problems in children with skeletal unloading. The aim of this study was to evaluate the effect of menatetrenone on bone metabolism in long-term glucocorticoid-treated children with alfacalcidol treatment. Twenty children who had been treated with fixed dosages of prednisolone and alfacalcidol (0.03 microg/kg/day) for 24 weeks were enrolled in a prospective pilot study, and assigned to receive alfacalcidol (0.03 microg/kg/day) or alfacalcidol (0.03 microg/kg/day) plus menatetrenone (approximately 2 mg/kg/day). Bone biochemical markers and bone mineral density (BMD) were measured at baseline and after the 12-week treatment. In the group receiving alfacalcidol plus menatetrenone, serum carboxylated osteocalcin (OC) (p =0.0022) and lumbar BMD (p=0.0029) increased and serum undercarboxylated OC (p=0.0004) decreased significantly in comparison to the group receiving alfacalcidol; further, the change of lumbar BMD showed an inverse correlation to the change of serum undercarboxylated OC (r=-0.744, p=0.0134) and positive correlations to the baseline values of bone turnover markers such as serum levels of intact OC, bone-specific alkaline phosphatase and type I procollagen carboxyl extension peptide and urinary levels of deoxypyridinoline and N-telopeptide of type I collagen. No adverse effect was observed. This is a small short-term study, but its results suggest that menatetrenone effectively and safely increases lumbar BMD probably through carboxylation of OC in long-term prednisolone-treated children with alfacalcidol treatment who have a high bone turnover. Randomized double-blind controlled trials are needed to confirm our findings. PMID- 11403097 TI - Severe hypercholesterolemia in a double heterozygote for lipoprotein lipase deficiency (LPL(Arita)) and apolipoprotein epsilon4: a report of a family with LPL(Arita). AB - Although heterozygous lipoprotein lipase (LPL) deficiency is not rare, only part of the phenotypes may have been reported in Japan. Here we describe a Japanese family with LPL(Arita), the most common mutation linked to familial LPL deficiency in Japan, and show for the first time a heterozygote for the mutation who had marked hypercholesterolemia due to increased low-density lipoprotein (LDL) cholesterol. The proband's mother, one of the heterozygotes for LPL(Arita) in the family, had both severe hypercholesterolemia (total cholesterol 306 mg/dl) with an especially increase in LDL-cholesterol and mild hypertriglyceridemia (180 mg/dl). She had normal LDL receptor activity and did not show clear evidence of possible causes of secondary hyperlipidemia. In addition to being heterozygous for LPL deficiency, she was also heterozygous for apo epsilon4. Because the epsilon4 allele is known to be associated with higher LDL-cholesterol, heterozygous apo epsilon4 may be one of causes of her LDL-cholesterol elevation. The other three heterozygotes for LPL(Arita) were moderate drinkers, and all of them had both remarkable hypertriglyceridemia and mild hypercholesterolemia due to increased very-low-density lipoproteins (VLDL). The results suggest that heterozygotes for LPL(Arita) can exhibit various phenotypes of hyperlipidemia, that is, hypertrigliceridemia and/or hypercholesterolemia due to not only increased VLDL but also increased LDL. The phenotypes appear to depend on some other genetic and environmental factors. PMID- 11403098 TI - Increased ACTH levels do not alter renal 11beta-hydroxysteroid dehydrogenase type 2 gene expression in the sheep. AB - The regulation of renal 11beta-hydroxysteroid dehydrogenase type 2 (11betaHSD2) gene expression is poorly understood. Inhibition of expression can result in hypertension. An example of this is in ectopic adrenocorticotropin (ACTH) syndrome (EAS). Inhibition of 11betaHSD2 activity is suggested by the observed increased ratio of cortisol to cortisone in both plasma and urine. To investigate whether ACTH or ACTH-dependent steroids can modulate renal 11betaHSD2 gene expression we analysed renal 11betaHSD2 mRNA levels after treatment with ACTH of 1 H and 24 H and demonstrated no change in the levels of gene expression. We have demonstrated in this study that the expression of 11betaHSD2 in the kidney is unaltered by ACTH. The reduced inactivation of cortisol by 11betaHSD2 observed in EAS is likely to be in part due to end product inhibition or substrate overload of the enzyme by endogenous substrates (cortisol, corticosterone, etc) rather than inhibition of 11betaHSD2 at the transcriptional level by either ACTH or ACTH regulated steroids. PMID- 11403099 TI - Ectopic adrenocorticotropin syndrome exhibiting paradoxical adrenocorticotropin responsiveness to gonadotropin-releasing hormone. AB - In a 37-year-old man who had Cushing's syndrome, investigations, including overnight dexamethasone suppression test, corticotropin-releasing hormone (CRH) test, pituitary MRI and inferior petrosal sinus sampling suggested the presence of ectopic adrenocorticotropin (ACTH) production. Interestingly, gonadotropin releasing hormone (GnRH) increased plasma ACTH from 73 pg/ml to 708 pg/ml at 15 min. Furthermore, desmopressin also increased plasma ACTH whereas CRH and thyrotropin-releasing hormone (TRH) had no effect. Such paradoxical responses of plasma ACTH were observed repeatedly. A thoracic CT scan revealed a right anterior mediastinal mass, which was surgically resected. Histological and immunohistochemical examination confirmed that the tumor was an ACTH-producing carcinoid. ACTH and cortisol decreased immediately following surgery. Neither desmopressin nor GnRH administration resulted in elevation of plasma ACTH while ACTH-responsiveness to dexamethasone and CRH was restored. To our knowledge, this is the first report documenting GnRH responsiveness in ectopic ACTH syndrome. PMID- 11403100 TI - Plasma angiotensin II, renin activity and serum angiotensin-converting enzyme activity in non-insulin dependent diabetes mellitus patients with diabetic nephropathy. AB - The renin-angiotensin system (RAS) has been unequivocally implicated as a mediator of diabetic complications. The present study was designed to evaluate the RAS in non-insulin dependent diabetic patients with diabetic nephropathy. Plasma renin activity, plasma angiotensin II and serum angiotensin-converting enzyme (ACE) activity were measured in 45 non-insulin dependent diabetes mellitus (NIDDM) patients and 15 healthy non-diabetic controls. Diabetics were subdivided into 15 normoalbuminuric NIDDM subjects, 15 NIDDM patients with microalbuminuria and 15 diabetics with macroalbuminuria. Mean plasma renin activity for macroalbuminuric diabetics (0.65+/-0.10 ng/ml/hr) was significantly reduced than the controls (1.28+/-0.37 ng/ml/hr) (P<0.001), the diabetic group with microalbuminuria (1.08+/-0.48 ng/ml/hr) (P<0.05) and normoalbuminuric patients (1.56+/-0.82 ng/ml/hr) (P<0.001). A significant negative correlation was obtained between serum creatinine and plasma renin activity (r=-0.842, p<0.001) in macroalbuminuric NIDDM patients. Plasma angiotensin II was significantly decreased in non-complicated diabetics compared to healthy controls (4.36+/-1.49 pg/ml vs 14.87+/-3.48 pg/ml respectively, p<0.001). Non-insulin dependent diabetic patients with nephropathy had significantly higher plasma angiotensin II levels (28.99+/-5.88 pg/ml) than non-complicated diabetics (p<0.001). Serum ACE activity was increased in 53.3% of NIDDM patients. All diabetic groups showed increased serum ACE activity (normoalbuminuric NIDDM 114.9+/-28.3 nmol/min/ml, microalbuminuric NIDDM 127.9+/-31.2 nmol/min/ml and macroalbuminuric NIDDM 127.0+/-29.3 nmol/min/ml) when compared to the normal control group (76.3+/-16.5 nmol/min/ml) (p<0.001). No significant difference in serum ACE activity was obtained between normoalbuminuric and nephropathic diabetics or between diabetics with and without retinopathy. No significant correlation was obtained between serum ACE activity and blood pressure, blood glucose level and duration of diabetes. Thus plasma renin activity is decreased in diabetic nephropathy and negatively correlates with serum creatinine. Plasma angiotensin II is decreased in normoalbuminuric diabetics and elevated in diabetic nephropathy. Serum ACE activity is raised in NIDDM patients with no relation to albumin excretion rate. The role of increased ACE activity in NIDDM remains to be established. PMID- 11403101 TI - Sex difference in platelet aggregation detected by new aggregometry using light scattering. AB - Earlier studies in platelet aggregation have shown that females seemed to have greater aggregability than males as detected by conventional aggregometry which used light transmission (LT), but controversy still remains. This study was performed to determine whether sex difference exists in platelet aggregation by using the recently developed laser light scattering (LS) method, which can detect small aggregates (i.e., two or three platelets). Blood was drawn from healthy volunteers (10 male and 10 female in follicular phase after menstruation), and platelet aggregation was detected by either LT or LS method in platelet rich plasma. Platelet aggregation was stimulated by increasing concentration of adenosine 5'-diphosphate (ADP, 0, 0.5, 1 and 2 microM). To detect the effect of sex hormones, platelets were incubated with estradiol (10 nM) or testosterone (40 nM) for 30 min, then platelet aggregation studies were performed. LT method revealed that female had greater aggregability than male. With weak stimuli (< or = 1 microM ADP), LS method showed that females had more medium aggregates than males, and that testosterone decreased small aggregates, and that estradiol decreased all sizes of aggregates. These data suggest that the female is more conductive to platelet aggregation than the male at a physiologic concentration of ADP (< or = 1 microM), but that both estradiol (10 nM) and testosterone (40 nM) have countereffects on platelet aggregation at the same condition. Therefore, the reason why females have greater aggregability than males may partly be explained by their lack of testosterone, but the mechanism still remains to be elucidated. PMID- 11403102 TI - Adrenal insufficiency after incomplete resection of pituitary macrocorticotropinoma of Cushing's disease: role of high molecular weight ACTH. AB - A 15-year-old girl with Cushing's disease exhibited adrenal insufficiency following incomplete trans-sphenoidal resection of a large pituitary corticotropinoma, approximately 35 mm in diameter. Within two weeks following surgery, her plasma ACTH level decreased from 42 to 13 pmol/l, while, her plasma cortisol levels and urinary excretion of free cortisol decreased from 607 nmol/l and 1112 nmol/day to 94 nmol/l and 55 nmol/day, respectively. Immunoreactive ACTH was characterized in plasma using Sephadex G-75 column chromatography and measuring ACTH with immunoradiometric assay (IRMA) and radioimmunoassay (RIA) to determine additional peaks, other than the one demonstrated for 1-39 ACTH. In particular, when measured with RIA, a broad peak including the high molecular weight ACTH was detected as well as 1-39 ACTH. The bioactivity of the high molecular weight ACTH in patient plasma was lower than the reference range of 1 39 ACTH, which is determined by the ability of dispersed rat adrenocortical cells to secrete corticosterone. The large pituitary corticotropinoma found in this patient secreted not only 1-39 ACTH but also high molecular weight proopiomelanocortin (POMC)-derived peptides, which could be detected by measuring with IRMA and RIA for ACTH. Based on the results of biological activity and molecular ratios, no positive evidence could be found to support the hypothesis that the high molecular weight ACTH induced any postoperative adrenal insufficiency in this patient. However, based on this study, the possibility of adrenal insufficiency should be carefully monitored, even when post-operative remnant tumor tissue is clearly present in patients with Cushing's disease, accompanied by macrocorticotropinoma. PMID- 11403103 TI - Postoperative plasma cortisol levels predict long-term outcome in patients with Cushing's disease and determine which patients should be treated with pituitary irradiation after surgery. AB - Transsphenoidal surgery is the treatment of choice for ACTH-producing pituitary adenoma (Cushing's disease) and pituitary irradiation is widely considered the most appropriate treatment for patients with Cushing's disease for whom transsphenoidal surgery has been unsuccessful. We studied 49 consecutive patients who underwent transsphenoidal surgery for the treatment of Cushing's disease at Tokyo Women's Medical University from 1977-1997 with a mean follow-up duration of 87.6 months (range, 24-253 months). We examined the relationship between postoperative endocrinological data, assessed between 3 and 8 weeks after surgery, and long-term outcome and efficacy of pituitary irradiation after surgery. Long-term remission was defined as the regression of the symptom and signs of Cushing's syndrome, and restoration of normal levels of plasma ACTH, cortisol and urinary free cortisol, together with adequate suppression of morning plasma cortisol levels following the administration of low dose (1 mg) of dexamethasone. Thirty patients had no additional treatment after pituitary surgery. Only 1 of 25 patients (4%) whose postoperative plasma cortisol level was less than 2 microg/dl developed recurrent disease whereas 3 out of 5 patients with postoperative plasma cortisol levels higher than 2 microg/dl relapsed. Postoperative external pituitary radiation was used to treat the remaining 19 patients. Four patients who received radiation therapy had a low or undetectable postoperative plasma cortisol level (<2 microg/dl, 56 nmol/L) and all of these patients developed hypopituitarism whereas 5 patients with subnormal plasma cortisol levels (2.0-10.0 microg/dl) remained in remission. Among 10 patients with persistent disease after surgery, 6 entered remission 6-47 months after irradiation but one of them subsequently relapsed after 108 months. These results suggest that 1) additional therapy should be avoided in patients with a postoperative plasma cortisol less than 2 microg/dl because relapse is very rare and radiotherapy will frequently induce hypopituitarism, 2) patients with a subnormal cortisol level following surgery should be treated with pituitary irradiation, because the relapse rate is reportedly high and radiotherapy is effective in preventing relapse, 3) radiotherapy in patients with persistent disease after surgery is effective only in 50% (5/10) of the patients. PMID- 11403104 TI - High prevalence of vitamin D deficiency in Japanese female patients with Graves' disease. AB - We reported previously that vitamin D deficiency is a causal mechanism of postoperative tetany in patients with Graves' disease. The aim of the present study was to determine the prevalence of vitamin D deficiency by reviewing serum 25(OH)D levels in 208 patients with Graves' disease (146 women, 62 men) during a 1 year period. Serum 25(OH)D levels were significantly lower (p < 0.001) in female Graves' patients (31.8 +/- 13.3 nmol/l) than in male patients (41.3 +/- 15.0 nmol/l). Vitamin D deficiency (defined as a serum 25(OH)D value below 25 nmol/l) was found in 40% of female patients and in 18% of male patients (p < 0.005). There was a significant seasonal variation in the 25(OH)D concentrations in female patients [amplitude 6.38 (95% CI, 5.42-7.56)], with values below 25 nmol/l found in 58% of female patients during the winter months. There were significant (p < 0.001) differences in serum 25(OH)D levels between age groups in the female patients. The concentrations were lowest in patients in their twenties (25.1 +/- 8.2 nmol/l) and highest in patients in their fifties and sixties (43.2 +/- 13.7 nmol/l). Serum 25(OH)D concentrations might be monitored in patients with Graves' disease during antithyroid drug therapy, and vitamin D and/or calcium supplements are recommended for patients with vitamin D deficiency. PMID- 11403105 TI - Prolactin binding analysis and immunohistochemical localization of prolactin receptor in porcine ovarian cells. AB - In the present study we searched for prolactin receptor (PRL-R) in porcine ovarian theca tissue (Tc) of small, medium and large follicles, as well as in early corpus luteum (ECL). The objectives of this investigation were: 1) comparison of the direct effect of PRL action on progesterone (P4) and estradiol (E2) secretion from Tc and ECL cells in culture with adequate effects caused by luteinizing hormone (LH). 2) detection of the presence and distribution of PRL-R in thecal tissue of porcine follicles and in ECL. Tissues were cultured as monolayers either in control M199 medium with calf serum or in medium either with PRL (100 ng/ml) or with LH (100 ng/ml). After 2 days in vitro cultured media were assayed for steroid concentrations by radioimmunoassays. Content and distribution of PRL-R were evaluated by Scatchard analysis and by an immunohistochemical assay. Separated theca layers as well as fragments of ECL were excised on dry ice, homogenized, and incubated with [125I]-PRL. PRL stimulated P4 secretion from Tc 10-fold versus controls. LH stimulated P4 secretion only 2.5-fold. E2 secretion was stimulated by PRL 2.7-fold and by LH 2.4-fold. LH enhanced P4 secretion from ECL cells by 18% while PRL increased P4 secretion by as much as 73%. Femtomol amounts of PRL-R protein were detected in theca tissues of medium and large follicles and also in ECL, which was in accordance with immunohistochemical results. The results showed for the first time the presence of PRL-R in porcine Tc and ECL. PMID- 11403106 TI - Clinical significance of the insulin resistance index as assessed by homeostasis model assessment. AB - To examine the clinical significance of the insulin resistance index as determined by homeostasis model assessment (HOMA-IR), we investigated the relationship between HOMA-IR and the insulin resistance estimated by the euglycemic-hyperinsulinemic clamp method in various subgroups and compared the significance of HOMA-IR with that of fasting plasma insulin levels (FIRI). HOMA IR was significantly correlated to the inverse of the glucose infusion rate (1/GIR) in both diabetic and non-diabetic subjects (r=0.747, P<0.0001 and r=0.419, P<0.002, respectively). In the diabetic patients, treatment with sulfonylureas did not weaken this correlation (r=0.833, P<0.0001). HOMA-IR was found to be closely related to FIRI (r=0.932, P<0.0001), but HOMA-IR was more closely associated with 1/GIR than FIRI was. HOMA-IR as well as 1/GIR was correlated with the visceral fat area (VFA) more closely than with the subcutaneous fat area (SFA), while FIRI was correlated almost equally with both of them. In conclusion, HOMA-IR is a convenient and beneficial method for evaluating insulin resistance, especially in subjects with visceral fat accumulation, and reflects insulin resistance obtained by euglycemic clamp more accurately than FIRI alone. PMID- 11403107 TI - Clinical implication of serial leptin measurement in subjects with type 2 diabetes mellitus. AB - Plasma leptin concentration is closely associated with body fat in humans, with energy restriction inducing a greater decrease in plasma leptin than in body fat. Since adequate energy restriction is mandatory in diet therapy of diabetes mellitus especially in obese subjects, the present study was undertaken to evaluate the clinical implication of serial leptin measurement in the management of diabetic patients. Fifty-four consecutive subjects with type 2 diabetes, who were subjected to adjusted energy restriction during hospitalization, were enrolled in the study. During their hospitalization period (24+/-4 days), plasma leptin concentrations decreased from 6.9+/-0.7 to 5.7+/-0.6 microg/l (P<0.0001) in the overall subjects, and the %change in plasma leptin (-13.9%) was greater than the %changes in body mass index (BMI) and percent body fat (-1.7% and -4.7%, respectively). The %change in plasma leptin was positively correlated with the %changes in BMI and plasma C-peptide (r=0.526, P<0.0001 and r=0.446, P<0.002, respectively) and negatively with a %change in plasma ketone bodies (r=-0.516, P<0.005). Multiple regression analysis revealed that the %changes in BMI and plasma C-peptide were independent determinants of the %change in plasma leptin. In addition, 38 subjects were followed up after discharge. Three months after discharge, plasma leptin concentrations significantly increased by 25.6%, which was again much greater than the %change in BMI (+0.9%). In 28 subjects who showed increase in plasma leptin levels after discharge, BMI was also increased. In contrast, the remaining 10 subjects without the increase in plasma leptin kept their BMI unchanged. Throughout the observation period, the changes in plasma leptin were prominent in the subjects with BMI greater than 25 kg/m2. In conclusion, plasma leptin concentrations showed greater changes than the alterations in anthropometric indexes during the observation period. Serial leptin measurement may be useful to estimate adherence to energy restriction especially in obese subjects with type 2 diabetes. PMID- 11403108 TI - Immunohistochemical localization of somatostatin receptor type 2A in rat and human tissues. AB - In this study, we elucidated the cellular localization of somatostatin receptor (SSTR) by immunohistochemistry using an antibody specific for SSTR type 2A (SSTR2A) in various organs of rat and human. SSTR2A expression was basically similar in rat and human, except in the pancreas and adrenal cortex. In the pituitary gland, the posterior lobe and the majority of growth hormone cells and some ACTH and TSH cells expressed SSTR2A. In rat adrenal gland, the zona glomerulosa strongly expressed SSTR2A, whereas zone-specific immunoreactivity was not observed in human. The adrenal medulla moderately expressed SSTR2A in both rat and human. SSTR2A immunoreactivity was observed in islet cells and some ductal cells in human pancreas, and also in acinar cells of rat pancreas. In gastrointestinal (GI) tract, the majority of crypt cells and nerve plexuses strongly expressed SSTR2A. The number of SSTR2A positive cells was much more than that of chromogranin A positive endocrine cells. In the kidney, the glomerular capillaries and collecting tubules, but not proximal tubules, showed immunoreactivity. SSTR2A immunoreactivity was observed not only in endocrine cells but also in non-endocrine cells. PMID- 11403109 TI - A systematic review of physical interventions for patellofemoral pain syndrome. AB - OBJECTIVE: Physical interventions (nonpharmacological and nonsurgical) are the mainstay of treatment for patellofemoral pain syndrome (PFPS). Physiotherapy is the most common of all physical interventions and includes specific vastus medialis obliquus or general quadriceps strengthening and/or realignment procedures (tape, brace, stretching). These treatments appear to be based on sound theoretical rationale and have attained widespread acceptance, but evidence for the efficacy of these interventions is not well established. This review will present the available evidence for physical interventions for PFPS. DATA SOURCES: Computerized bibliographic databases (MEDLINE, Current Contents, CINAHL) were searched, including the keywords "patellofemoral," "patella," and "anterior knee pain," combined with "treatment," "rehabilitation," and limited to clinical trials through October 2000. STUDY SELECTION: The critical eligibility criteria used for inclusion were that the study be a controlled trial, that outcome assessments were adequately described, and that the treatment was a nonpharmacological, nonsurgical physical intervention. RESULTS: Of the 89 potentially relevant titles, 16 studies were reviewed and none of these fulfilled all of the requirements for a randomized, controlled trial. Physiotherapy interventions were evaluated in eight trials, and the remaining eight trials examined different physical interventions. Significant reductions in PFPS symptoms were found with a corrective foot orthosis and a progressive resistance brace, but there is no evidence to support the use of patellofemoral orthoses, acupuncture, low-level laser, chiropractic patellar mobilization, or patellar taping. Overall the physiotherapy interventions had significant beneficial effects but these interventions were not compared with a placebo control. There is inconclusive evidence to support the superiority of one physiotherapy intervention compared with others. CONCLUSIONS: The evidence to support the use of physical interventions in the management of PFPS is limited. There appears to be a consistent improvement in short-term pain and function due to physiotherapy treatment, but comparison with a placebo group is required to determine efficacy, and further trials are warranted for the other interventions. PMID- 11403110 TI - Lateral plantar nerve entrapment in a competitive gymnast. PMID- 11403111 TI - Blood testing in sports: hematological profile of a convicted athlete. PMID- 11403112 TI - The spontaneous healing of a torn anterior cruciate ligament. PMID- 11403113 TI - Epiphyseal injuries of the proximal phalanx of the hallux. PMID- 11403114 TI - Effects of testosterone precursor supplementation on intensive weight training. PMID- 11403115 TI - Reducing the progression of disability in older adults. PMID- 11403116 TI - Do textbooks agree on their advice on ice? AB - OBJECTIVE: To study ice therapy guidance in sports medicine textbooks. DATA SOURCES: A systematic search of a convenience sample of textbooks. STUDY SELECTION: 45 general sports medicine texts were included in the study. DATA EXTRACTION: The indices and chapter headings of each text were searched using key words "ice," "cryotherapy," "soft tissue injury," "muscle," and "bruise." DATA SYNTHESIS: In 17 of the textbooks, there was no guidance on the duration, frequency, or length of ice treatment or on the use of barriers between ice and the skin. Advice on treatment duration was given in 28 texts but recommendations differed depending on the particular ice therapy, injury location, or severity. There was considerable variation in the recommended duration and frequency of advised treatments. CONCLUSION: There was little guidance in the standard textbooks on ice application, and the advice varied greatly. There is a need for evidence-based sport and exercise medicine with a consensus on the appropriate use of ice in acute soft tissue injury. PMID- 11403117 TI - Stress fracture sites related to underlying bone health in athletic females. AB - OBJECTIVE: The study tested the hypothesis that females who sustain stress fractures of cancellous bone have decreased bone density. DESIGN: A retrospective, controlled, cross-sectional study. SETTING: The setting of the study was a tertiary care center for Women's Sports Medicine. PATIENTS: 20 female patients under the age of 40 who had suffered a stress fracture and who had a positive diagnostic study (radiograph, bone scan, or magnetic resonance imaging) were included in the study. INTERVENTIONS: Patients who had a positive diagnostic study (radiograph, bone scan, or magnetic resonance imaging) for the diagnosis of stress fracture also underwent dual energy X-ray absorptiometry (DEXA) scans. MAIN OUTCOME MEASURE: Bone density measured by the DEXA scan, as defined by the World Health Organization criteria for osteopenia (greater than one standard deviation from the standard age-matched control). RESULTS: 8 of 9 patients with cancellous stress fractures had DEXA scans indicating osteopenia while only 3 of 11 patients with stress fractures of cortical bone had a scan indicating osteopenia (p = 0.01). CONCLUSIONS: A cancellous stress fracture in a female may be a warning sign of early onset osteopenia. We recommend that young females who have documented stress fractures of cancellous bone or cortical bone (with risk factors for osteopenia) undergo bone density evaluation. PMID- 11403118 TI - Sport and recreation-related head injuries treated in the emergency department. AB - OBJECTIVE: Head injury (HI) in sport is common and can have serious consequences. This study examines the epidemiology of sport/recreational (SR)-related HI presenting to the emergency department (ED). DESIGN: Retrospective review of medical records. SETTING: Five EDs in the Capital Health Region (Edmonton) located in the province of Alberta, Canada. PATIENTS: All persons in a 1-year period reporting to the EDs with an HI. HI was defined as IC9-CM coded skull fracture, loss of consciousness, or concussion. MAIN OUTCOME MEASURES: Hospitalization, utilization of diagnostic testing, and discharge destination. RESULTS: In total, 10,877 (3%) of 288,948 ED encounters were for sports and recreational injuries; 358 (3%) were for HI. Males (71%) were more frequently injured; patients < 20 years old were involved in 66% of all HI cases. The highest proportion of HI occurred during ice hockey (21%), cycling (13%), and playground-related activities (8%). 9% of HI were hospitalized (versus 4% admission rate for other SR injuries: p < 0.01). CONCLUSIONS: These results demonstrate the utility of an ED-based injury registry and indicate that patients with HI presenting to the ED from SR activities are common. These injuries appear to be more severe than other types of SR injuries treated in the ED. PMID- 11403119 TI - An evaluation of head movement in backboard-immobilized helmeted football, lacrosse, and ice hockey players. AB - OBJECTIVE: Improper handling of an unstable neck injury in the prehospital setting may result in iatrogenically induced neurologic injury. Due to helmet design, stabilization of the cervical spine in American football does not require routine removal of the helmet and shoulder pads prior to transport. Adequate data is not available evaluating hockey and lacrosse helmets. This study compares the amount of head movement in American football, lacrosse, and ice hockey helmets during head and neck stabilization procedures. STUDY DESIGN: Prospective. PARTICIPANTS: 12 ice hockey, 9 football, and 9 lacrosse athletes from an National Collegiate Athletic Association Division 1 program. SETTING: On-campus biomechanical laboratory with three HiRes cameras, routinely tested for accuracy. METHODS: Athletes were immobilized on backboards as per protocol. Three motion analysis HiRes cameras follow retroreflective markers placed on the helmet and bite mouthplate to measure relative head and helmet motion. MAIN OUTCOME MEASURES: Helical angles determine the relative range of motion of the head inside the helmets. RESULTS: The mean range of head motion for football players was 4.88 degrees (n = 9, SD 2.07), lacrosse players 6.56 degrees (n = 9, SD 1.61), and ice hockey players 5.54 degrees (n = 12, SD 1.19). These results were not significantly different (p > 0.05). CONCLUSIONS: The rotational head motion seen inside standard immobilized lacrosse and ice hockey helmets is similar to that seen in football helmets. This supports the safety of prehospital stabilization of the potential cervical spine-injured ice hockey and lacrosse athletes with in-line stabilization and helmet in place. Extrapolation of data may not be applicable to other helmet designs, and future studies are needed to determine the safety of emergency procedures in all helmet designs. PMID- 11403120 TI - The effects of creatine dietary supplementation on anterior compartment pressure in the lower leg during rest and following exercise. AB - OBJECTIVE: To examine the effects of creatine supplementation on anterior compartment pressure of the lower leg at rest and following exercise. DESIGN: 14 college-age males received creatine or placebo supplementation for 34 days. At baseline, anterior compartment pressure was measured preexercise, immediately postexercise, and 1, 5, and 15 minutes postexercise after a level treadmill run for 20 minutes at 80% of maximal aerobic power. INTERVENTION: Following baseline testing, subjects began a 6-day creatine or placebo loading phase at a dosage of 0.3 g x kg body mass(-1) x d(-1). This was followed by a 28-day maintenance phase at a dosage of 0.03 g x kg body mass(-1) x d(-1). Subjects and investigators were blinded as to treatment administration. Subjects continued to exercise during the supplementation period. After 6 days and 34 days of supplementation. anterior compartment pressure was measured at rest and following exercise. RESULTS: Creatine supplementation for 6 days significantly increased compartment pressure compared with the placebo group at rest (76%), immediately post- (150%), 1 minute post- (125%), 5 minute post- (106%), and 15 minute postexercise (77%). Anterior compartment pressures continued to remain significantly higher for the creatine group compared with the placebo group at rest (72%), immediately post- (125%) and 1 minute postexercise (180%) after 34 days of creatine supplementation. CONCLUSIONS: These data indicate that creatine supplementation abnormally increases anterior compartment pressure in the lower leg at rest and following 20 minutes of level running at 80% of maximal aerobic power. PMID- 11403121 TI - Gender influences on career opportunities, practice choices, and job satisfaction in a cohort of physicians with certification in sports medicine. AB - OBJECTIVE: To examine the gender differences in practice patterns, experiences, and career opportunities for family physicians who practice sports medicine. DESIGN: Descriptive, self-administered questionnaire. PARTICIPANTS: Family physicians with Certificate of Added Qualification (CAQ) in sports medicine were surveyed. The survey was sent to all women with a CAQ in Sports Medicine and a random sample of 20% of the men with CAQs in sports medicine. MAIN OUTCOME MEASURE: Survey consisted of multiple choice, Likert scale, and opened-ended questions. The data was analyzed with contingency tables, with gender as the dependent variable. RESULTS: Response rate to the survey was 75%, which included 42 females and 102 males. Demographics of our population demonstrated some gender differences. Males were of higher average age (41.1 vs. 38.1), and more likely to be married and have children. Practice types, location, and time spent in sports medicine did not differ with the exception of training room and event coverage. Males were more likely to cover all levels of training room except at the Division I level, where the percent of males and females covering training rooms were equal. Males were also more likely to cover all types of sporting events. Job satisfaction and reasons for choosing current jobs did not show significant gender differences. However, factors affecting career opportunities did vary. Professional relationships with athletic trainers and coaches were perceived to be different by males and females surveyed. CONCLUSIONS: Our survey of sports medicine physicians showed some gender differences in practice patterns relative to training room and sporting event coverage. Surprisingly, there were not many differences in the factors that affected job choice and factors affecting job opportunities with the exception of gender itself. However, our study does not conclude how or when gender begins to affect the female sports medicine physician's career opportunities. PMID- 11403122 TI - Modeling the survival of Escherichia coli O157:H7 in uncooked, semidry, fermented sausage. AB - The data collected from studies to monitor inactivation of Escherichia coli O157:H7 in uncooked fermented salami were used to develop models to describe survival of the organism. Three models were developed that included different variables to best describe E. coli O157:H7 reduction. Model A included the variables water activity (a(w)), pH, time, and quadratic variables pH and time. Model B separated the processing stages into fermentation and drying. The fermentation included the variables pH and temperature x time (ttarea) and interaction between the two variables. The drying stage was modeled using the variables time and a(w) and interaction between the two. Model C looked at variables a(w) and the time at pH 5.3 to achieve a 2-D log reduction of E. coli O157:H7 and the interaction between the variables. The variables selected for inclusion in all the models were significant at the P < 0.0001 level. The predicted values for all models correlated well to the observed values with R2 of 0.888, 0.828, 0.836, and 0.818 for models A. Bferm, Bdrying, and C, respectively. The models were validated using data from a trial not used to develop the model. In general the predicted reduction in E. coli O157:H7 count in uncooked fermented salami was greater than for the observed E. coli O157:H7 log reductions for all models, but the relation between the two was linear. The results demonstrate that modeling can be a useful tool in assessing manufacturing practices for uncooked fermented salami processes. PMID- 11403123 TI - Sublethal sanitizer stress and adaptive response of Escherichia coli O157:H7. AB - The effect of sublethal exposure to peroxyacetic acid (PAA) sanitizer on adaptation to peroxidative stress and development of thermal cross-resistance was investigated in Escherichia coli O157:H7. Acute sublethal PAA sanitizer exposure was used to represent a contact scenario. Cultures were grown in Trypticase soy yeast extract broth. Acute treatment cultures were pretreated with 0.1% PAA, then all cultures were challenged at either 80 mM H202 or 54 degrees C. Acute and peroxide control cultures showed substantially increased peroxidative tolerance (D80mM > 2 h) versus negative control cultures not exposed to sanitizer (D80mM = 0.19+/-0.03 h). The inactivation rate of the acetic acid control (D80mM = 0.21+/ 0.05 h) was similar to the negative control rate. Acute (D54 degrees C = 0.55+/ 0.07 h) cultures did not exhibit increased thermal resistance versus the control (D54 degrees C = 0.54+/-0.07 h). Thermal injury was determined as difference in D54 degrees C value (deltaD54 degrees c) obtained on pyruvate and deoxycholate media. Thermal-induced injury was not observed in either control (deltaD54 degrees C = 0.04 h) or acute (deltaD54 degrees C = 0.05 h) cultures. PMID- 11403124 TI - Survival and death of Salmonella typhimurium and Campylobacter jejuni in processing water and on chicken skin during poultry scalding and chilling. AB - Salmonella Typhimurium and Campylobacter jejuni were inoculated in scalding water, in chilled water, and on chicken skins to examine the effects of scalding temperature (50, 55, and 60 degrees C) and the chlorine level in chilled water (0, 10, 30, and 50 ppm), associated with the ages of scalding water (0 and 10 h) and chilled water (0 and 8 h), on bacterial survival or death. After scalding at 50 and 60 degrees C, the reductions of C. jejuni were 1.5 and 6.2 log CFU/ml in water and <1 and >2 log CFU/cm2 on chicken skins; the reductions of Salmonella Typhimurium were <0.5 and >5.5 log CFU/ml in water and <0.5 and >2 log CFU/cm2 on skins, respectively. The age of scalding water did not significantly (P > 0.05) affect bacterial heat sensitivity. However, the increase in the age of chilled water significantly (P < 0.05) reduced the chlorine effect. In 0-h chilled water. C. jejuni and Salmonella Typhimurium were reduced by 3.3 and 0.7 log CFU/ml, respectively, after treatment with 10 ppm of chlorine and became nondetectable with 30 and 50 ppm of chlorine. In 8-h chilled water, the reduction of C. jejuni and Salmonella Typhimurium was <0.5 log CFU/ml with 10 ppm of chlorine and ranged from 4 to 5.5 log CFU/ml with 50 ppm of chlorine. Chlorination of chilled water did not effectively reduce the bacteria attached on chicken skins. The D-values of Salmonella Typhimurium and C. jejuni were calculated for the prediction of their survival or death in the poultry scalding and chilling. PMID- 11403125 TI - Inactivation of Escherichia coli O1 57:H7, Listeria monocytogenes, and Lactobacillus leichmannii by combinations of ozone and pulsed electric field. AB - Pulsed electric field (PEF) and ozone technologies are nonthermal processing methods with potential applications in the food industry. This research was performed to explore the potential synergy between ozone and PEF treatments against selected foodborne bacteria. Cells of Lactobacillus leichmannii ATCC 4797, Escherichia coli O157:H7 ATCC 35150, and Listeria monocytogenes Scott A were suspended in 0.1% NaCl and treated with ozone, PEF, and ozone plus PEE Cells were treated with 0.25 to 1.00 microg of ozone per ml of cell suspension, PEF at 10 to 30 kV/cm, and selected combinations of ozone and PEF. Synergy between ozone and PEF varied with the treatment level and the bacterium treated. L. leichmannii treated with PEF (20 kV/cm) after exposure to 0.75 and 1.00 microg/ml of ozone was inactivated by 7.1 and 7.2 log10 CFU/ml, respectively; however, ozone at 0.75 and 1.00 microg/ml and PEF at 20 kV/cm inactivated 2.2, 3.6, and 1.3 log10 CFU/ml, respectively. Similarly, ozone at 0.5 and 0.75 microg/ml inactivated 0.5 and 1.8 log10 CFU/ml of E. coli, PEF at 15 kV/cm inactivated 1.8 log10 CFU/ml, and ozone at 0.5 and 0.75 microg/ml followed by PEF (15 kV/cm) inactivated 2.9 and 3.6 log10 CFU/ml, respectively. Populations of L. monocytogenes decreased 0.1, 0.5, 3.0, 3.9, and 0.8 log10 CFU/ml when treated with 0.25, 0.5, 0.75, and 1.0 microg/ml of ozone and PEF (15 kV/cm), respectively; however, when the bacterium was treated with 15 kV/cm, after exposure to 0.25, 0.5, and 0.75 microg/ml of ozone, 1.7, 2.0, and 3.9 log10 CFU/ml were killed, respectively. In conclusion, exposure of L. leichmannii, E. coli, and L. monocytogenes to ozone followed by the PEF treatment showed a synergistic bactericidal effect. This synergy was most apparent with mild doses of ozone against L. leichmannii. PMID- 11403126 TI - Survival of enterohemorrhagic Escherichia coli O157:H7 in retail mustard. AB - Escherichia coli O157:H7 survival in acid foods such as unpasteurized apple cider and fermented sausage is well documented. Researchers have determined that E. coli O157:H7 can survive in refrigerated acid foods for weeks. The potential of acid foods to serve as a vector of E. coli O157:H7 foodborne illness prompted this study to determine the fate of this organism in retail mustard containing acetic acid when stored at room and refrigerated temperatures. Various retail brands of dijon, yellow, and deli style mustard, pH ranging from 3.17 to 3.63, were inoculated individually with three test strains of E. coli O157:H7. Samples were inoculated with approximately 1.0 x 10(6) CFU/g, incubated at room (25+/-2.5 degrees C) and refrigerated (5+/-3 degrees C) temperatures, and assayed for surviving test strains at predetermined time intervals. An aliquot was appropriately diluted and plated using sorbitol MacConkey agar (SMAC). When the test strain was not recoverable by direct plating, the sample was assayed by enrichment in modified tryptic soy broth and recovered using SMAC. Growth of E. coli O157:H7 test strains was inhibited in all retail mustard styles. E. coli O157:H7 was not detected in dijon style mustard beyond 3 h at room and 2 days at refrigerated temperatures. Survival in yellow and deli style mustard was not detected beyond 1 h. Overall, test strain survival was greater at refrigerated than room temperature. Retail mustard demonstrated the ability to eliminate effectively any chance contamination by this organism within hours to days, suggesting that these products are not a likely factor in E. coli O157:H7 foodborne illness. PMID- 11403127 TI - Evaluation of a polymerase chain reaction-based system for detection of Salmonella enteritidis, Escherichia coli O157:H7, Listeria spp., and Listeria monocytogenes on fresh fruits and vegetables. AB - A polymerase chain reaction (PCR)-based detection system, BAX, was evaluated for its sensitivity in detecting Salmonella Enteritidis, Escherichia coli O157:H7, Listeria sp., and Listeria monocytogenes on fresh produce. Fifteen different types of produce (alfalfa sprouts, green peppers, parsley, white cabbage, radishes, onions, carrots, mushrooms, leaf lettuce, tomatoes, strawberries, cantaloupe, mango, apples, and oranges) were inoculated, in separate studies, with Salmonella Enteritidis, E. coli O157:H7, and L. monocytogenes down to the predicted level of 1 CFU per 25-g sample. Detection by BAX was compared to recovery of the inoculated bacteria by culture methods according to the Food and Drug Administration's (FDA) Bacteriological Analytical Manual (BAM). BAX was essentially as sensitive as the culture-based method in detecting Salmonella Enteritidis and L. monocytogenes and more sensitive than the culture-based method for the detection of E. coli O157:H7 on green pepper, carrot, radish, and sprout samples. Detection of the pathogenic bacteria in samples spiked with a predicted number of less than 10 CFU was possible for most produce samples, but both methods failed to detect L. monocytogenes on carrot samples and one of two mushroom and onion samples spiked with less than 100 CFU. Both BAX and the culture method were also unable to consistently recover low numbers of E. coli O157:H7 from alfalfa sprouts. The PCR method allowed detection of Salmonella Enteritidis, E. coli O157:H7, and L. monocytogenes at least 2 days earlier than the conventional culture methods. PMID- 11403128 TI - Elimination of fecal coliforms and F-specific RNA coliphage from oysters (Crassostrea virginica) relaid in floating containers. AB - Declining oyster (Crassostrea virginica) production in the Chesapeake Bay has stimulated aquaculture based on floats for off-bottom culture. While advantages of off-bottom culture are significant, the increased use of floating containers raises public health and microbiological concerns, because oysters in floats may be more susceptible to fecal contamination from storm runoff compared to those cultured on-bottom. We conducted four commercial-scale studies with market-size oysters naturally contaminated with fecal coliforms (FC) and a candidate viral indicator, F-specific RNA (FRNA) coliphage. To facilitate sampling and to test for location effects, 12 replicate subsamples, each consisting of 15 to 20 randomly selected oysters in plastic mesh bags, were placed at four characteristic locations within a 0.6- by 3.0-m "Taylor" float, and the remaining oysters were added to a depth not exceeding 15.2 cm. The float containing approximately 3,000 oysters was relaid in the York River, Virginia, for 14 days. During relay, increases in shellfish FC densities followed rain events such that final mean levels exceeded initial levels or did not meet an arbitrary product end point of 50 FC/100 ml. FRNA coliphage densities decreased to undetectable levels within 14 days (16 to 28 degrees C) in all but the last experiment, when temperatures fell between 12 and 16 degrees C. Friedman (nonparametric analysis of variance) tests performed on FC/Escherichia coli and FRNA densities indicated no differences in counts as a function of location within the float. The public health consequences of these observations are discussed, and future research and educational needs are identified. PMID- 11403129 TI - Use of spent irrigation water for microbiological analysis of alfalfa sprouts. AB - Numerous outbreaks of foodborne illness have been linked to the consumption of raw sprouts. Sprout producers have been advised by the Food and Drug Administration to include microbiological testing of spent irrigation water during production as part of an overall strategy to enhance the safety of sprouts. Alfalfa sprouts and irrigation water were analyzed to show the feasibility of using irrigation water for monitoring the microbiological safety of sprouts. Sprouts and water were produced and harvested from both commercial scale (rotary drum) and consumer-scale (glass jars) equipment. Rapid increases of aerobic mesophiles occurred during the first 24 h of sprouting, with maximum levels achieved after 48 to 72 h. The counts in irrigation water were on average within approximately 1 log of their respective counts in the sprouts. Similar results were obtained for analysis of Escherichia coli O157:H7 in irrigation water and sprouts grown from artificially inoculated seeds. Testing of spent irrigation water indicated the contamination status of alfalfa sprouts grown from seeds associated with outbreaks of Salmonella infection. PMID- 11403130 TI - Continuous online processing of fecal- and ingesta-contaminated poultry carcasses using an acidified sodium chlorite antimicrobial intervention. AB - The objective of this study was to determine the effectiveness of the combined use of an inside-outside-bird-washer for the removal of visible contamination and an online acidified sodium chlorite (ASC) spray system in reducing microbial levels on contaminated poultry carcasses. Specifically, we attempted to determine if this technique (referred to as continuous online processing [COP]) would (i) eliminate the need for offline reprocessing of contaminated carcasses, (ii) meet Zero Fecal Tolerance standards, and (iii) attain significant reductions in titers of some of the commonly found bacterial species. Carcasses were sampled for Ercherichia coli, Salmonella, and Campylobacter at five stations along the processing lines in a series of five commercial plant studies to compare the efficacy of the COP system to that of offline processing. The microbiological quality of fecally contaminated carcasses was found to be significantly better following COP treatment (E. coli, 0.59 log10 CFU/ml; Salmonella, 10.0% incidence) than after standard offline reprocessing (E. coli, 2.37 log10 CFU/ml; Salmonella, 31.6% incidence). Zero Fecal Tolerance requirements were met by all but 2 (0.2%) of the 1.127 carcasses following COP. COP also significantly reduced the titers of Campylobacter; residual titers were 1.14 log10 CFU/ml (49.1% incidence) following COP, compared to 2.89 log10 CFU/ml (73.2% incidence) in carcasses that underwent offline reprocessing. These results support the combined use of an inside-outside-bird-washer for the removal of visible contamination and an online ASC spray system to reduce microbial levels in commercially processed poultry. PMID- 11403131 TI - Bioluminescence ATP assay for estimating total plate counts of surface microflora of whole cantaloupe and determining efficacy of washing treatments. AB - The surface microflora of cantaloupes were estimated using a bioluminescence ATP assay, and results were compared to plate count data. Cantaloupes were treated as follows: (i) water washed, or (ii) washed in solutions of sodium hypochlorite (1,000 mg/liter) or hydrogen peroxide (5%) for 5 min. Bioluminescence ATP assay results showed differences in ATP level/cm2 of cantaloupes dipped in chlorine or hydrogen peroxide solution; ATP levels in these washed samples were lower than in controls due to antimicrobial action of the treatments on the cantaloupe surface. Linear correlations were found between the bioluminescence ATP assay and aerobic plate counts of unwashed cantaloupe (r2 = 0.995) and those washed with water (r2 = 0.990) determined before storage. Lower correlations between the bioluminescence ATP assay and the aerobic plate counts were observed on cantaloupes stored for 120 h at 20 degrees C (r2 = 0.751) than at 4 degrees C (r2 = 0.980) without washing treatment. Lower correlation at 20 degrees C may be the result of clusters or growth that occurred in chains. ATP levels of washed cantaloupes correlated well with bacterial plate counts (r2 = 0.999). A reliable minimum detectable threshold using the bioluminescence ATP assay was established at 3 log10 fg/cm2 corresponding to 4 log10 CFU/cm2. Bioluminescence ATP assay is not recommended for washed samples where the microbial load is near or below the threshold. Therefore, the bioluminescence ATP assay will be recommended for quick estimation of total microbial load on cantaloupe surfaces where the population is expected to exceed this threshold. The assay can save the industry time by eliminating the required incubation required by the conventional methods. PMID- 11403132 TI - Microbiological meat quality in high- and low-capacity slaughterhouses in Sweden. AB - The purpose of this study was to investigate the bacterial count on beef and pork carcasses, a comparison being made between different-sized slaughterhouses in Sweden. Samples were taken from the flank and sternum of 200 beef and 200 pork carcasses, with half of the samples being collected from low-capacity slaughterhouses. Sampling was carried out at the end of the slaughter-line from 25 beef carcasses and/or 25 pork carcasses in each abattoir. Analyses were performed of the aerobic microorganisms, coliform bacteria, coagulase-positive staphylococci, and presumptive Escherichia coli. No significant differences were found in the amount of aerobic microorganisms between pork carcasses from low- and high-capacity slaughterhouses. In beef carcasses, however, there was a significantly greater amount of aerobic microorganisms in beef carcasses slaughtered at low-capacity slaughterhouses. Within the group of high-capacity abattoirs there was a very small variation in the amount of aerobic bacteria between the different slaughterhouses that could be explained by their having almost the same evisceration technique. The evisceration technique differs more among the low-capacity slaughterhouses, which is probably the main reason for the wide variation in the amount of aerobic bacteria. Coliform bacteria, coagulase positive staphylococci, and presumptive E. coli were more common on pork carcasses than on beef carcasses. There were also significantly higher amounts of these bacteria in pork carcasses from the high-capacity slaughterhouses. PMID- 11403133 TI - Traditional versus hazard analysis and critical control point-based inspection: results from a poultry slaughter project. AB - Federal meat and poultry inspection has changed little since the Federal Meat Inspection Act was passed in 1906, followed by the Poultry Products Inspection Act of 1957 and related amendments. These acts mandate sensory or organoleptic (sight, smell, and touch) inspection of all carcasses. For several decades, the U.S. Department of Agriculture's Food Safety and Inspection Service (FSIS) has been urged by various organizations to move to a scientific, risk-based inspection system. In partial response to these calls, the FSIS has developed new slaughter inspection models that are currently being tested with volunteer plants in the hazard analysis and critical control point (HACCP)-based inspection models project. To evaluate whether plants operating under the new inspection models perform at least as well as they did under the current or traditional system, microbial and organoleptic data are being collected before and after the implementation of the new inspection models. In this article, we describe the baseline and models data collection procedures and present the results of the baseline and models data collection for eight plants that slaughter young chickens. The results from the first eight volunteer plants suggest that inspection under the new models is equivalent and in some ways superior to that of traditional inspection. This pilot project suggests that new slaughter inspection systems, which rely on HACCP principles with FSIS oversight and verification services, can maintain or even improve food safety and other consumer protection conditions relative to traditional hands-on inspection methods. PMID- 11403134 TI - Primers and a specific DNA probe for detecting lactic acid bacteria producing 3 hydroxypropionaldehyde from glycerol in spoiled ciders. AB - Of the 40 strains isolated from several spoiled ciders where glycerol was degraded, 36 were identified as Lactobacillus collinoides, three were Lactobacillus hilgardii, and one was Lactobacillus mali. However, only 30 L. collinoides and two L. hilgardii could degrade glycerol. The glycerol dehydratase activity was shown. The main product of the transformation was 1.3 propanediol. Two DNA primers GD1 and GD2 were chosen in the region encoding one of the subunits of glycerol dehydratase of Citrobacter freundii, Klebsiella pneumoniae, Klebsiella oxytoca, Salmonella Typhimurium, and Clostridium pasteurianum. A 279 bp amplicon in polymerase chain reaction amplification was obtained with the genomic L. collinoides IOEB 9527 DNA as template. The amino acid sequence deduced from the amplicon DNA sequence showed a very high similarity and identity with the gene of gram-negative and C. pasteurianum species. After labeling, the amplicon was used as DNA probe in dot-blot hybridization with the genomic DNA of all the tested strains. Only strains that could degrade glycerol hybridized. Moreover, polymerase chain reactions using GDI and GD2 revealed only glycerol dehydratase genes of positive L. collinoides and L. hilgardii strains. The primers and the amplicon proved to be suitable and reliable tools to detect the lactic acid bacteria involved in the deterioration of cider. PMID- 11403135 TI - Inhibition of growth of nonproteolytic Clostridium botulinum type B in sous vide cooked meat products is achieved by using thermal processing but not nisin. AB - The safety of refrigerated processed foods of extended durability (REPFEDs) with respect to nonproteolytic Clostridium botulinum is under continuous evaluation. In the present study, mild (P7.0(85.0) values 0 to 2 min [P, pasteurization value; z-value 7.0 degrees C; reference temperature 85.0 degrees C]) and increased (P7.0(85.0) values 67 to 515 min) heat treatments were evaluated in relation to survival of nonproteolytic C. botulinum type B spores in sous vide processed ground beef and pork cubes. The use of two concentrations of nisin in inhibition of growth and toxin production by nonproteolytic C. botulinum in the same products was also evaluated. A total of 96 samples were heat processed and analyzed for C. botulinum by BoNT/B gene-specific polmerase chain reaction and for botulinum toxin by a mouse bioassay after storage of 14 to 28 days at 4 and 8 degrees C. Predictably, after mild processing all samples of both products showed botulinal growth, and one ground beef sample became toxic at 8 degrees C. The increased heat processing, equivalent to 67 min at 85 degrees C. resulted in growth but not toxin production of C. botulinum in one ground beef sample in 21 days at 8 degrees C: in the pork cube samples no growth was detected. The increased heating of both products resulted in higher sensory quality than the milder heat treatment. Nisin did not inhibit the growth of nonproteolytic C. botulinum in either product; growth was detected in both products at 4 and 8 degrees C, and ground beef became toxic with all nisin levels within 21 to 28 days at 8 degrees C. Aerobic and lactic acid bacterial counts were reduced by the addition of nisin at 4 degrees C. The study demonstrates that the mild processing temperatures commonly employed in sous vide technology do not eliminate nonproteolytic C. botulinum type B spores. The intensity of each heat treatment needs to be carefully evaluated individually for each product to ensure product safety in relation to nonproteolytic C. botulinum. PMID- 11403136 TI - Glove barriers to bacterial cross-contamination between hands to food. AB - Human hands are an important source of microbial contamination of foods. However, published data on the effectiveness of handwashing and glove use in a foodservice setting are limited. Bacterial transfer through foodservice quality gloves was quantified using nalidixic acid-resistant Enterobacter aerogenes (a nonpathogenic surrogate with attachment characteristics similar to Salmonella). Five transfer rates were determined: chicken to bare hand, chicken to hand through gloves, bare hand to lettuce, hand to lettuce through gloves (with low inoculum on hands), and hand to lettuce through gloves (with high inoculum on hands). At least 30 observations were made for each percent transfer rate using 30 individual volunteers. The logarithm of percent transfer data were then fit to distributions: chicken to bare hand, normal (0.71, 0.42); chicken to hand through gloves, gamma (5.91, 0.40, -5.00); bare hand to lettuce, logistic (1.16, 0.30); hand to lettuce through gloves (low inoculum), normal (0.35, 0.88); hand to lettuce through gloves (high inoculum), normal (-2.52, 0.61). A 0.01% transfer was observed from food to hands and from hands to food when subjects wore gloves and a 10% transfer was observed without a glove barrier. These results indicate that gloves are permeable to bacteria although transfer from hands to food through a glove barrier was less than without a glove barrier. Our results indicate that gloves may reduce both bacterial transfer from food to the hands of foodservice workers and in subsequent transfer from hands back to food. PMID- 11403137 TI - Changes in phosphoglyceride composition during storage of ultrahigh-temperature milk, as assessed by 31P-nuclear magnetic resonance: possible involvement of thermoresistant microbial enzymes. AB - Soluble phosphoglycerides were studied in ultrahigh-temperature (UHT) milk by 31P nuclear magnetic resonance. It was shown that, during storage of UHT milk, manufactured from raw milk with poor microbial quality, glycerophosphocholine and glycerophosphoethanolamine disappeared in parallel with an increase in alpha glycerophosphate (GP). Storage at 10, 20, and 30 degrees C showed a faster transformation as the temperature increased. UHT milk samples manufactured from raw milks with better microbial quality and submitted to severe heat processes did not display changes in phosphoglycerides during storage. Screening of commercial UHT milks showed variations regarding the presence of GP, while in pasteurized milk samples, the appearance of GP occurred when the commercial life had been exceeded. Inoculation of sterile milk with Pseudomonas fluorescens NCIB9046 and incubation at 10 degrees C supported that changes in phosphoglycerides could be the consequence of a phosphodiesterase activity of bacterial origin, able to survive UHT processing. A similar behavior was observed between this activity and proteolytic activity. The potential application of the detection of these compounds as spoilage predictor indices is discussed. PMID- 11403138 TI - Viability, efficacy, and storage stability of freeze-dried biocontrol agent Candida sake using different protective and rehydration media. AB - Viability, efficacy against Penicillium expansum on Golden Delicious apples, and storage stability of freeze-dried Candida sake strain CPA-1 were studied. The effect of several protective agents and rehydration media was investigated in the freeze drying of C. sake. Skimmed milk at 10% concentration was a good rehydration medium for all protectants tested. In general, good viability results were obtained when the same solution was used as a protectant and as a rehydration medium. The best survival was obtained when C. sake cells were protected with 10% lactose + 10% skimmed milk and rehydrated with skimmed milk (85% viability). The potential for biocontrol of the best freeze-dried treatments against P. expansum on apples was compared with that of fresh cells. Freeze-dried treatments at 1 x 10(7) CFU/ml reduced the incidence of decay by 45 to 66%. The best biocontrol effect was obtained with cells that had been freeze dried using 10% lactose + 10% skimmed milk as a protectant and 1% peptone as a rehydration medium, with a 66% reduction in rot incidence. However, in all treatments, the efficacy of freeze-dried cells was significantly lower than fresh cells. The stability of freeze-dried samples decreased during storage and was influenced by storage temperature. In the best treatment, storage of C. sake cells for 60 days at 4 degrees C resulte in final concentrations of 2.5 x 10(8) CFU/ml, which was a 10-fold reduction in relation to the initial starting concentration of cells prior to freeze drying. PMID- 11403139 TI - Presence of Escherichia coli O157:H7 in ground beef and ground baby beef meat. AB - A total of 114 beef and baby beef samples were examined. The samples included ground baby beef, mixed ground baby beef and pork, and chopped and shaped meat. The samples were analyzed from 30 different grocery stores in Zagreb, Croatia. The object of this study was to evaluate the prevalence of Escherichia coli O157:H7 in the samples that can enhance the potential risk of outbreaks of hemorrhagic colitis and hemolytic uremic syndrome. The results in all tested samples of E. coli O157:H7 were negative. A single sample was positive in a latex agglutination test using antiserum to O157:H7. It was identified as Proteus vulgaris at the Pasteur Institute, Paris, France. This result correlates positively with cross-contamination with Yersinia enterocolitica 09, Brucella abortus, Salmonella type N, and Pseudomonas maltophila. PMID- 11403140 TI - Multiplication in egg yolk and survival in egg albumen of Salmonella enterica serotype Enteritidis strains of phage types 4, 8, 13a, and 14b. AB - Refrigeration of eggs is vital for restricting the multiplication of Salmonella enterica serotype Enteritidis contaminants, but differences between Salmonella Enteritidis strains or phage types in their survival and multiplication patterns in egg contents might influence the effectiveness of refrigeration standards. The present study compared the abilities of 12 Salmonella Enteritidis isolates of four phage types (4, 8, 13a, and 14b) to multiply rapidly in egg yolk and to survive for several days in egg albumen. The multiplication of very small numbers of Salmonella Enteritidis inoculated into yolk (approximately 10(1) CFU/ml) was monitored during 24 h of incubation at 25 degrees C, and the survival of much larger numbers of Salmonella Enteritidis inoculated into albumen (approximately 10(5) CFU/ml) was similarly evaluated during the first 3 days of incubation at the same temperature. In yolk, the inoculated Salmonella Enteritidis strains multiplied to mean levels of approximately 10(3) CFU/ml after 6 h of incubation and 10(8) CFU/ml after 24 h. In albumen, mean levels of approximately 10(4) CFU/ml or more of Salmonella Enteritidis were maintained through 72 h. Although a few differences in multiplication and survival were observed between individual isolates, the overall range of values was relatively narrow, and no significant differences (P < 0.05) were evident among phage types. PMID- 11403141 TI - Citrus juice composition does not influence radiation sensitivity of Salmonella enteritidis. AB - Food substrate chemistry is known to influence radiation sensitivity of pathogenic bacteria. The sensitivity of a citrus juice outbreak strain of Salmonella Enteritidis to gamma radiation was determined in five commercial orange juice formulations. The juices differed in pH (3.87 to 4.13), calcium concentration (2.1 versus 36.9 mM), juice composition (orange versus orange tangerine blend), and antioxidant power (11,751 to 12,826 microM ferric reducing antioxidant power units). The Dgamma (dose required to achieve 90% destruction) varied only slightly (0.35 to 0.37 kGy), with no significant (P < 0.05) differences among any of the suspending juices. These results indicate that Salmonella Enteritidis sensitivity to gamma radiation is not strongly influenced by the composition of formulated commercial orange juices. PMID- 11403142 TI - Validation of a 5-log10 reduction of Listeria monocytogenes following simulated commercial processing of Lebanon bologna in a model system. AB - Recently, numerous product recalls and one devastating outbreak that claimed 21 lives were attributed to Listeria monocytogenes in ready-to-eat meat products. Consequently, the Food Safety and Inspection Service published a federal register notice requiring manufacturers of ready-to-eat meat and poultry products to reassess their hazard analysis and critical control point plans for these products as specified in 9 CFR 417.4(a). Lebanon bologna is a moist, fermented ready-to-eat sausage. Because of undesirable quality changes. Lebanon bologna is often not processed above 48.9 degrees C (120 degrees F). Therefore, the present research was conducted to validate the destruction of L. monocytogenes in Lebanon bologna batter in a model system. During production, fermentation of Lebanon bologna to pH 4.7 alone significantly reduced L. monocytogenes by 2.3 log10 CFU/g of the sausage mix (P < 0.01). Heating the fermented mix to 48.9 degrees C in 10.5 h destroyed at least 7.0 log10 CFU of L. monocytogenes per g of sausage mix. A combination of low pH (5.0 or lower) and high heating temperatures (> or =43.3 degrees C, 115 degrees F) destroyed more than 5 log10 CFU of L. monocytogenes per g of sausage mix during the processing of Lebanon bologna. In conclusion, an existing commercial process, which was validated for destruction of Escherichia coli O157:H7, was also effective for the destruction of more than 5 log10 CFU of L. monocytogenes. PMID- 11403143 TI - Closed-circuit system for the depuration of mussels experimentally contaminated with hepatitis A virus. AB - In Italy, the consumption of raw or slightly cooked mussels represents the most important risk factor for the transmission of hepatitis A virus (HAV). Although there exist effective methods for the bacterial depuration of contaminated mussels, these methods are poorly effective on enteric viruses. The objective of the present study was to evaluate the effectiveness of a closed-circuit depuration system that uses both ozone and UV light for disinfecting water and that allows salinity and temperature, important parameters for the metabolism of mussels (Mytilus galloprovincialis), to be maintained at constant levels. The results showed that this depuration method decreased the viral load (from 1.72 log 50% tissue culture infective dose [TCID50] ml(-1) to <1 log TCID50 ml(-1) within 24 h and from 3.82 log TCID50 ml(-1) to <1 log TCID50 ml(-1) within 48 h). However, in both cases, after 120 h of depuration, a residual amount of virus capable of replicating in cells was detected. These results show that depuration, even if performed with advanced systems, may not guarantee the absence of virus. PMID- 11403144 TI - Survival of poliovirus in New Zealand green-lipped mussels, Perna canaliculus, on refrigerated and frozen storage. AB - Poliovirus survival in live and frozen mussels during storage was assessed by both viral culture and molecular methods. Live New Zealand green-lipped mussels were incubated overnight at 20 degrees C in an aerated tank of filtered seawater seeded with the poliovirus 2 (PV2) vaccine strain. An extraction and concentration method that preserved viral infectivity was used to recover PV2 taken up by the mussels at day 0, at day 2 after storage at 4 degrees C, and at days 7, 14, and 28 after storage at -20 degrees C. This method allowed both culture and molecular analysis to be carried out. Presence of intact PV2 in each batch of mussels was determined by a pan-enterovirus specific reverse transcription-polymerase chain reaction (RT-PCR) and confirmed by dot-blot hybridization. Survival of infectious PV2 was determined by the monolayer plaque assay. After 48 h at 4 degrees C, infectious PV2 levels were 81% of the original level detected in the mussels. Infective virus levels then declined to 66, 53, and 44% after storage at -20 degrees C for 7, 14, and 28 days, respectively. Generic RT-PCR methods were 10 times more sensitive than cell culture techniques for virus detection but did not give information on virus infectivity. The survival of infectious pathogenic viruses in fresh and frozen mussels on storage constitutes a potential health risk and so is a major concern for public health authorities. PMID- 11403145 TI - Antimicrobial effects of corn zein films impregnated with nisin, lauric acid, and EDTA. AB - Bacterial growth during food transport and storage is a problem that may be addressed with packaging materials that release antimicrobials during food contact. In a series of five experiments, EDTA, lauric acid (LA), nisin, and combinations of the three antimicrobial agents were incorporated into a corn zein film and exposed to broth cultures of Listeria monocytogenes and Salmonella Enteritidis for 48 h (sampled at 2, 4, 8, 12, 24, and 48 h). Four experiments used starting cultures of 10(8) CFU/ml in separate experiments tested against each bacterium; the fifth experiment examined the inhibitory effect of selected antimicrobial agents on Salmonella Enteritidis with an initial inoculum of 10(4) CFU/ml. L. monocytogenes cell numbers decreased by greater than 4 logs after 48 h of exposure to films containing LA and nisin alone. No cells were detected for L. monocytogenes (8-log reduction) after 24-h exposure to any film combination that included LA. Of all film agent combinations tested, none had greater than a 1-log reduction of Salmonella Enteritidis when a 10(8)-CFU/ml broth culture was used. When a 10(4) CFU/ml of Salmonella Enteritidis initial inoculum was used, the films with EDTA and LA and EDTA, LA, and nisin were bacteriostatic. However, there was a 5-log increase in cells exposed to control within 24 h. The results demonstrate bacteriocidal and bacteriostatic activity of films containing antimicrobial agents. PMID- 11403146 TI - Vitamin B1 and B6 retention in milk after continuous-flow microwave and conventional heating at high temperatures. AB - The effect of continuous-flow microwave treatment at high temperatures on the retention of vitamins B1 and B6 in raw milk with different fat content was evaluated. Results were compared with those obtained using a conventional system (tubular heat exchanger) with the same heating and cooling phases. Heat treatment of whole (3.4% fat) and skim (0.5% fat) milk at 90 degrees C produced no losses of vitamin B1 or vitamin B6 (pyridoxamine and pyridoxal). However, at 110 and 120 degrees C, while vitamin B1 content of milk remained constant, pyridoxamine increased (4 to 5% and 9 to 11%, respectively) and pyridoxal decreased (5 to 6% and 9 to 12%, respectively). Under the assayed conditions, no differences were observed between the content of these vitamins in conventionally and continuous flow microwave-treated milk. PMID- 11403147 TI - Detection of hazelnut proteins in foods by enzyme immunoassay using egg yolk antibodies. AB - An enzyme immunoassay (EIA) was developed for the detection of hazelnut proteins in foods. This assay used inexpensive chicken egg yolk antibodies in a sandwich EIA format for the immunospecific capture and detection of hazelnut proteins present in a variety of different food matrices. The assay was able to detect less than 1 ppm of hazelnut protein in most of the foods tested and did not exhibit any appreciable cross-reactivity with other nuts or food matrices. This assay will be a useful tool for the food industry and regulatory agencies that wish to test foods for the presence of undeclared hazelnut allergens. PMID- 11403148 TI - Foodborne illness outbreaks in Korea and Japan studied retrospectively. AB - The average prevalence of reported foodborne illness from 1981 to 1995 was 2.44 per 100,000 population in Korea, and 28.01 in Japan. The mean case fatality rate in Korea was 0.74% and in Japan, 0.03%. When both prevalence and case fatality rates in Korea and Japan were compared during the same period, the prevalence in Japan was much higher than that in Korea. However, the case fatality rate of patients in Korea was much higher than that in Japan. The distribution of monthly and seasonal patterns of foodborne illness outbreaks strongly indicate the outbreaks may be associated with climatic conditions, frequencies of national holidays, and vacation seasons. Comparison study indicates that the foodborne illness outbreaks in Korea most frequently involved homemade foods (47% of the total cases); in Japan, restaurants accounted for 31.3%. Foodborne illness cases of bacterial origin in Korea were 59.3% of the total and included Salmonella spp. (20.7%). Vibrio (17.4%), Staphylococcus (9.7%), pathogenic Escherichia coli (2.4%), and other species (9.1%); in Japan, 72.8% of the total cases and the majority of the bacterial foodborne illness was caused by Vibrio (32.3%), Staphylococcus (15.9%), Salmonella (14.2%), pathogenic E. coli (3.0%), and other species (7.2%). In conclusion, the outbreaks of foodborne illness in Korea and Japan may be mainly caused by improper food handling, and their occurrences may be differentiated according to food sources. PMID- 11403149 TI - Current importance of ochratoxin A-producing Aspergillus spp. AB - Ochratoxin A (OA) is receiving attention worldwide because of the hazard it poses to human and animal health. OA contamination of commodities, such as cereals or pork and poultry meat, is well recognized. Nevertheless, there is an increasing number of articles reporting OA contamination in other food commodities, such as coffee, beer, wine, grape juice, and milk, in the last few years. This continuous and increasing exposure to OA that humans experience is reflected in the high incidence of OA in both human blood and milk in several countries. OA was believed to be produced only by Aspergillus ochraceus and closely related species of section Circumdati and by Penicillium verrucosum; however, in the genus Aspergillus, the production of OA has been recently reported by species outside the section Circumdati. Thus, it has been clearly established as a metabolite of different species of the section Nigri, such as Aspergillus niger and Aspergillus carbonarius. OA production ability by Aspergillus spp. is more widespread than previously thought; therefore, there is the possibility that unexpected species can be new sources of this mycotoxin in their natural substrates. PMID- 11403150 TI - Can the physician predict the neuropathologist? PMID- 11403151 TI - Role of type 10 17beta-hydroxysteroid dehydrogenase in the pathogenesis of Alzheimer's disease. PMID- 11403152 TI - The spectrum of vascular disease in dementia. From ischaemia to amyloid angiopathy. PMID- 11403153 TI - Transgenic mouse models of cerebral amyloid angiopathy. PMID- 11403154 TI - Alpha-synuclein. Axonal transport, ligand interaction and neurodegeneration. PMID- 11403155 TI - Dementia with Lewy bodies. PMID- 11403156 TI - Mouse models of alpha-synucleinopathy and Lewy pathology. Alpha-synuclein expression in transgenic mice. PMID- 11403157 TI - Ubiquitin and the molecular pathology of neurodegenerative diseases. AB - Ubiquitin plays a central role in normal cellular function as well as in disease. It is possible to group ubiquitin-immunostained structures into several main groups, the most distinctive being the ubiquitin/intermediate filament/alphaB crystallin family of inclusions that seem to represent a general cellular response to abnormal proteins recently termed the aggresomal response. While ubiquitin immunohistochemistry is a very useful technique for detecting pathological changes and inclusion bodies in the nervous system this alone is not enough to classify inclusions, and a panel of antibodies is recommended to clarify any findings made by screening tissues with anti-ubiquitin. Several mechanistic possibilities now exist to explain the accumulation of ubiquitinated proteins in cells of the nervous system, understanding of which should lead to new therapeutic advances in the group of chronic neurodegenerative diseases. PMID- 11403158 TI - Perisomatic granules of hippocampal CA1 neurons in Alzheimer's disease, pre Alzheimer stage and Pick's disease: an overlooked pathological entity. PMID- 11403159 TI - Pathological mechanisms in polyglutamine expansion diseases. PMID- 11403160 TI - Tau gene mutations and tau pathology in frontotemporal dementia and parkinsonism linked to chromosome 17. PMID- 11403161 TI - Dementia in the neuronal ceroid-lipofuscinoses. PMID- 11403162 TI - Neurodegeneration-associated proteins and inflammation in sporadic inclusion-body myositis. PMID- 11403163 TI - Mitochondrial dysfunction in neurodegenerative disorders and ageing. PMID- 11403164 TI - Argyrophilic grain disease. A frequent dementing disorder in aged patients. PMID- 11403165 TI - The molecular parameters of tau pathology. Tau as a killer and a witness. PMID- 11403166 TI - Tau pathology in neurons and glial cells of aged baboons. PMID- 11403167 TI - Human tau transgenic mice. Towards an animal model for neuro- and glialfibrillary lesion formation. PMID- 11403168 TI - Regulation of alzheimer beta-amyloid precursor trafficking and metabolism. PMID- 11403169 TI - Myocardial viability assessment in regions of left ventricular dysfunction. Part I: Radionuclide imaging and clinical implications. PMID- 11403170 TI - Simple and reliable preparation of pentavalent 99Tcm-dimercaptosuccinic acid at alkaline pH without oxygen bubbling. AB - We investigated a simple and reliable method for the preparation of 99Tcm dimercaptosuccinic acid (99Tcm(V)-DMSA) without the addition of oxygen. The effect of pH, amount of reducing agent, and oxygen addition in the synthesis of 99Tcm(V)-DMSA were evaluated. At pH 9, we obtained a radiochemical yield of 95% +/- 1.2% within 10 min and a high stability until 7 h, with 92% +/- 1.5% radiochemical purity. However, at a pH lower than 9, the radiochemical yield was below 90% within 10 min, and a longer reaction time was needed to obtain a radiochemical yield above 90%. The addition of oxygen did not have an additional effect on the radiochemical yield or its stability at pH 9, whereas it increased the radiochemical yield of 99Tcm-(V)-DMSA at pH 7. It was noted that the smaller the amount of reducing agent used, the higher was the radiochemical yield obtained at pH 7. However, at pH 9, the radiochemical yield was not dependent on the amount of reducing agent. In conclusion, the synthesis of 99Tcm(V)-DMSA was more dependent on the pH of the reaction mixture than on the amount of reducing agent or the addition of oxygen. The adjustment of pH 9 was the easiest and most effective method for the synthesis of 99Tcm(V)-DMSA using a commercial kit for 99Tcm(III)-DMSA. PMID- 11403171 TI - Protein binding of 99Tcm-DTPA compared with other GFR tracers. AB - 99Tcm-Diethylenetriaminepentaacetic acid (99Tcm-DTPA) has been postulated to be more bound to plasma proteins than are other radiopharmaceuticals used for measurement of glomerular filtration rate (GFR). The results of protein binding experiments are, however, highly related to methodology. The aim of the present study was to compare the protein binding of the 99Tcm-DTPA preparation we use in our daily routine with that of the other five 99Tcm-DTPA preparations available and with other GFR tracers by using the same ultrafiltration method. Protein binding was studied in vitro after incubation in donor plasma and in vivo after single injection in patients. A correction for the uneven distribution of ultrafiltrable ions ('Donnan effect') was included. Quality control of six 99Tcm DTPA preparations performed with thin layer chromatography showed a high labelling efficiency (96.2-99.9%). The in vitro protein binding ranged from 9.25% to 11.12%. The in vitro protein binding of 99Tcm-DTPA was not significantly different from that of 51Cr-ethylenediaminetetraacetic acid (51Cr-EDTA) and 125I iothalamate (10.99% vs 12.15%, and 9.55%). There was no difference in the in vivo protein binding of 99Tcm-DTPA 5 and 40 min after injection (13.28% vs 12.58%), and there was no difference in the protein binding of 99Tcm-DTPA and 51Cr-EDTA (12.93% vs 12.54%). In conclusion, in vitro and in vivo protein binding of 99Tcm DTPA was 10-13%. The protein binding was not different from the binding of 51Cr EDTA and 125I-iothalamate. PMID- 11403172 TI - Does inadequate exercise lower the accuracy of myocardial perfusion scintigraphy? AB - The predictive accuracy of exercise myocardial perfusion scintigraphy (EMPS) in detecting coronary artery disease (CAD) in patients who fail to achieve an adequate level of exercise is not clear. This investigation was carried out in order to compare the sensitivity, specificity and accuracy of EMPS in adequate exercise patients with those in inadequate exercise patients. We have retrospectively compared the results of EMPS with coronary angiography (CAG). One hundred and forty-eight patients with both tests within 6 weeks were included. Adequate exercise was defined as > or = 85% maximally predicted heart rate for age. The overall sensitivity and specificity of EMPS to detect CAD were 92.5% (74/80) and 75%, (51/68), respectively. The sensitivity and specificity in adequate exercise patients were 94.1% (32/34) and 67.6% (23/34), whereas those in inadequate exercise patients were 91.3% (42/46) and 82.4% (28/34). The accuracy was 80.9% (55/68) and 87.5% (70/80), respectively. Patients with inadequate exercise had lower sensitivity but higher specificity of EMPS for detecting CAD, and achieved a higher accuracy than those with adequate exercise. PMID- 11403173 TI - Comparison of 99Tcm-MIBI with 201Tl chloride SPET in patients with malignant brain tumours. AB - The purpose of this study was to evaluate the usefulness of 99Tcm-MIBI accumulation for the differentiation of histological diagnosis of malignant brain tumours in comparison with the findings obtained using 201Tl chloride. A total of 25 patients with malignant brain tumours were investigated. The histological categories of tumours included glioblastoma multiforme (n = 5), anaplastic astrocytoma (n = 4), malignant lymphoma (n = 5), and metastatic tumour (n = 11). Simultaneous dual single photon emission tomography (SPET) images with 99Tcm-MIBI and 201Tl were acquired 15 min (early) and 2 h (delayed) after injection, and the early ratio, delayed ratio and retention index were measured. The new indices 201Tl/99Tcm-MIBI ratios and 201Tl/99Tcm-MIBI retention index were also calculated. With respect to the histological type, a higher retention index using 99Tcm-MIBI was noted in glioblastoma multiforme compared with metastatic tumour. Higher values of both ratios using 201Tl were noted in glioblastoma multiforme compared to metastatic tumour. The value of the delayed ratio obtained using 201Tl was higher in glioblastoma multiforme than in anaplastic astrocytoma, and the value was also higher in malignant lymphoma than in metastatic tumour. The 201Tl/99Tcm-MIBI early ratio of glioblastoma multiforme was significantly higher than that of metastatic brain tumour. The 201Tl/99Tcm-MIBI retention index of malignant lymphoma was significantly higher than that of glioblastoma multiforme. In the histological type of tumour, 99Tcm-MIBI is not superior to 201Tl, but the combined indices using 201Tl/99Tcm-MIBI may add new information about differential diagnosis. PMID- 11403174 TI - A rapid and stable ITLC procedure for the determination of the radiochemical purity of 99Tcm-tetrofosmin. AB - 99Tcm-tetrofosmin (Myoview, Amersham Healthcare) is widely used as a radiopharmaceutical for myocardial perfusion imaging. Control of its radiochemical purity after reconstitution is usually performed by means of ITLC SG paper chromatography in a mobile phase of methylene chloride/acetone (65/35), as recommended by the manufacturer. The present study describes the application of tetrahydrofuran in phosphate buffer for the development of 99Tcm-tetrofosmin on ITLC-SG strips. The use of this mobile phase significantly improves the separation between labelled tetrofosmin and unbound pertechnetate. The time for development is about 1 min and the solvent is stable for at least 1 year. In addition, the volume spotted on the strip does not affect the migration of 99Tcm tetrofosmin. The labelling efficiency of 99Tcm-tetrofosmin can successfully be monitored by means of this method as a daily routine procedure. PMID- 11403175 TI - Verapamil decreases accumulation of 99Tcm-MIBI and 99Tcm-tetrofosmin in human breast cancer and soft tissue sarcoma cell lines. AB - 99Tcm-methoxyisobutylisonitrile (99Tcm-MIBI) and 99Tcm-tetrofosmin are cationic tracers recognized by the efflux pump P-glycoprotein (Pgp). Verapamil has been shown to be a competitive inhibitor of Pgp, and was one of the first multidrug resistant reversing agents identified. The aim of this preclinical in vitro study was to evaluate the effects of verapamil on the accumulation of 99Tcm-MIBI and 99Tcm-tetrofosmin in the human breast cancer cell lines MCF-7 and SK-BR-3 and in the human soft tissue sarcoma cell lines SW 982 and SW 1353, in comparison with respective control cells, i.e. without preincubation with verapamil. After preincubation with 10 or 100 microM of verapamil for 15 or 30 min, the 99Tcm-MIBI and 99Tcm-tetrofosmin accumulation in cells was assessed at 10, 30 and 60 min after incubation with these tracers. Addition of verapamil caused a decline in the accumulation of the two tracers at all incubation times, as compared with control cells. These effects of verapamil were neither dose- nor preincubation time-dependent in most cells. Our data indicate that verapamil is not a promising agent for increasing the sensitivity of scintigraphy with 99Tcm-MIBI or 99Tcm tetrofosmin, or for evaluating Pgp tumour status in these types of tumours. PMID- 11403176 TI - Salivary scintigraphy for assessing the protective effect of pilocarpine in head and neck irradiated tumours. AB - Patients with head and neck cancers can develop salivary hypofunction after radiotherapy. The use of pilocarpine during radiotherapy treatment has been shown to be an effective treatment, although its usefulness is being discussed. The aim of this study was: (1) to determine the value of a semiquantitative scintigraphy method for measuring the uptake and excretory salivary function of patients with head and neck irradiated tumours; and (2) to study the usefulness of pilocarpine as a salivary gland protector during radiotherapy. We prospectively studied 49 patients (mean age 61 years, range 29-87 years) with head and neck cancer in need of radiotherapy. Patients were divided into two groups consecutively: group P (26 patients) received 5 mg of pilocarpine three times per day starting the day before radiation therapy, and group NP (23 patients) received radiotherapy without pilocarpine and were used as the control group. Salivary gland scintigraphy and a visual analogue scale (VAS) of mouth dryness were obtained from each patient before radiotherapy and during the first year after treatment. The most frequent finding after radiotherapy was a quick impairment in parotid and submaxillary excretion (P < 0.001). There were no statistical differences comparing the pilocarpine group against the non-pilocarpine group. Parotid and submaxillary uptake significantly decreased after radiotherapy in both groups (P < 0.001). However, a tendency to recover within the pilocarpine group was observed in both the parotids and the submaxillary glands at 12 months. No differences were found comparing the VAS results in both groups. Strikingly, VAS data did not correlate with salivary gland dysfunction observed by means of scintigraphy. In conclusion, salivary scintigraphy is a useful technique to evaluate objectively the salivary gland function of patients with head and neck irradiated tumours as well as to test the response to pilocarpine. However, despite better results on the salivary uptake at 12 months, pilocarpine did not significantly improve salivary gland function. PMID- 11403177 TI - Diagnostic accuracy of 99Tcm-HIDA with cholecystokinin and gallbladder ejection fraction in acalculous gallbladder disease. AB - The diagnostic accuracy and clinical impact of 99Tcm hepatic iminodiacetic acid (HIDA) imaging with cholecystokinin (CCK) was investigated in a prospective study of 359 patients over an 11 year period. All patients presented with right upper quadrant biliary type pain and had a normal ultrasound investigation prior to imaging. CCK was administered as a 3 min infusion at peak gallbladder uptake of HIDA. A gallbladder ejection fraction (GBEF) was used to quantify the gallbladder response to CCK. Two hundred and forty-four of 359 (68%) patients had an abnormal GBEF (< or = 35%). One hundred and thirty-four of 141 (95%) patients who underwent cholecystectomy had abnormal surgical/histological findings and/or relief of symptoms on long-term (mean 5.7 years) follow-up. Clinical follow-up, mean of 5.9 years, of the patients with GBEF > 35% showed 73/79 (92%) of them with little evidence of gallbladder disease. For a total 261 patients with mean clinical follow-up of 5.7 years the sensitivity of GBEF measurement is 95%, specificity is 92% and overall accuracy is 94%. It is concluded that 99Tcm-HIDA imaging, with a 3 min infusion of CCK, is a highly accurate technique and valuable in the diagnostic management of patients with suspected acalculous gallbladder disease. PMID- 11403178 TI - Assessment of regional myocardial wall motion and thickening by gated 99Tcm tetrofosmin SPECT: a comparison with magnetic resonance imaging. AB - Gated single photon emission computed tomography (SPECT) imaging allows the simultaneous assessment of both perfusion and function by using one single study. The assessment of regional wall motion and thickening pattern with gated SPECT allows viability studies to be performed. Magnetic resonance imaging (MRI) is well validated for the assessment of myocardial wall motion and thickening in patients with normal and impaired ventricular function. The aim of the study was to analyse the concordance between wall motion and thickening scores derived by gated SPECT and MRI imaging. Furthermore, the agreement for myocardial wall motion and thickening according to myocardial perfusion was analysed with both techniques. We studied a group of 21 patients, including 13 with a previous myocardial infarction (all more than 4 months before the study), using both gated SPECT 99Tcm-tetrofosmin myocardial perfusion imaging and MRI. A 13-segment model was used for both gated SPECT and MRI and each segment was visually scored using a scale of 1-3 for wall motion and thickening. There was a high agreement between gated SPECT and MRI for both wall motion (229/273, 84%; k = 0.72, P<0.001) and wall thickening (236/273, 86%; k = 0.77, P<0.001). The agreement for wall motion and thickening was 80% (k = 0.66) and 83% (k = 0.70), respectively, for patients with myocardial infarction; and 90% (k = 0.81) and 92% (k = 0.86), respectively (P = NS), for patients without myocardial infarction. Agreement in segmental wall motion and thickening scores between gated SPECT and MRI was 90% (k = 0.80) and 91% (k = 0.84), respectively, for segments with normal or mild to moderate hypoperfusion; and 71% (k = 0.45) and 77% (k = 0.57), respectively, for segments with severe hypoperfusion or no perfusion. Of the 70 (41%) segments that had severely diminished or no perfusion in post-myocardial infarction patients, 22 (31%) showed preserved wall motion and 17 (24%) showed preserved wall thickening both by gated SPECT and MRI, suggesting residual myocardial viability in malperfused segments. Our results suggest that gated SPECT imaging is a reliable tool for the assessment of regional wall motion and thickening in patients with known or suspected coronary artery disease. In patients with a previous myocardial infarction gated SPECT imaging has the potential to detect preserved wall motion and thickening in regions with fixed perfusion defects indicating the potential presence of residual myocardial viability. PMID- 11403179 TI - Radiopeptide transmitted internal irradiation of non-iodophil thyroid cancer and conventionally untreatable medullary thyroid cancer using. AB - AIM: Differentiated thyroid carcinomas (DTC) and medullary thyroid carcinomas (MTC) overexpress somatostatin receptor subtypes (sstr). The aim of this pilot study was to evaluate the tumour response of thyroid carcinomas to targeted irradiation with the radiolabelled somatostatin analogue [90Y]-1,4,7,10-tetra azacyclododecan-4,7,10-tricarboxy-methyl-1-yl-acetyl-D-Phe1-Tyr3-octreotide ([90Y]-DOTA-D-Phe1-Tyr3-octreotide, or 90Y-DOTATOC) which has a high affinity to subtype 2 and a low affinity to subtype 5. It shows no affinity to sstr1, sstr3 and sstr4. PATIENTS AND METHODS: Twenty patients (mean age 58 years; 50% female, 50% male) with thyroid cancer were included (medullary thyroid cancer (MTC), 12 patients; differentiated thyroid cancer (DTC), seven patients; papillar carcinoma (PC), four patients; follicular carcinoma (FC), three patients; anaplastic carcinoma (AC), one patient). All patients had been therapy resistant and had progressive disease before 90Y-DOTATOC therapy. The dose applied was between totals of 1700 MBq x m(-2) to 7400 MBq x m(-2) 90Y-DOTATOC, administered in one to four injections at intervals of 6 weeks. In the case of tumour progression under therapy, treatment was terminated. RESULTS: The overall antitumour effect (objective response and stable disease) was 35%; in MTC 42%, in DTC 29%, and in AC 0%. The objective overall response rate was 0%. A stable disease was achieved in 35% (7/20), and progressive disease was found in 65% (13/20). The median time to progression was 8 months, with a median follow-up of 15 months. The treatment was very well tolerated. There were no grade III/IV haematological or renal toxicities. CONCLUSION: Targeted radiotherapy using 90Y-DOTATOC is able to stop tumour progression in a small number of patients and therefore may be an alternative treatment option for resistant disease. More significant tumour responses in thyroid and medullary thyroid cancer may be obtained by using radiopeptides with pan-somatostatin characteristics. PMID- 11403180 TI - Correlation of 99mTc-sestamibi uptake with blood-pool and osseous phase 99mTc-MDP uptake in malignant bone and soft-tissue tumours. AB - Technetium-99m-sestamibi (99mTc-MIBI) imaging is a well-established modality in oncologic investigations. The current study aimed to investigate whether any relationship could be found between 99mTc-MIBI uptake and local perfusion in malignant bone and soft-tissue tumours. It also aimed to compare 99mTc-MIBI images with those of technetium-99m-methylene diphosphonate (99mTc-MDP) bone scintigraphy with regard to the activity distribution pattern, intensity and lesion extension. The study group included 24 patients with various bone and soft tissue tumours. Three-phase bone scintigraphy and 99mTc-MIBI studies were performed within the same week before any surgical and therapeutic intervention. Images were evaluated visually and quantitatively using regions of interest (ROIs) over the lesion and adjacent normal tissue. The 99mTc-MIBI study was positive with varying degrees of uptake (range, 1.4-5.3). The mean 99mTc-MIBI uptake and 99mTc-MDP blood-pool and osseous phase activity ratios were 2.5 +/- 0.5, 2.8 +/- 1.0 and 5.5 +/- 4.0, respectively. The correlation between the 99mTc MIBI uptake and blood-pool ratios was 0.70 (P<0.05). While activity distribution patterns were in agreement in 99mTc-MIBI and blood-pool images in the majority of cases, 99mTc-MIBI better delineated tumour viability and extension in five cases. In conclusion, 99mTc-MIBI accumulation shows a reasonable correlation with blood pool uptake assuming the presence of multifactorial mechanisms in addition to local hyperaemia. Better delineation of tumour outlines and cellular activity seems to be an advantage of 99mTc-MIBI scintigraphy which may be helpful in the evaluation of musculoskeletal tumours. PMID- 11403181 TI - Post-stress end-systolic left ventricular dilation: a marker of endocardial post ischemic stunning. AB - Several studies have shown the accuracy of gated single photon emission computed tomography (SPECT) using thallium-201 and technetium tracers in the assessment of myocardial perfusion and function. Gated SPECT has been successfully utilized to detect post-stress left ventricular ejection fraction (LVEF) reduction resulting from post-ischemic stunning in patients with coronary obstruction. The aim of this study was to evaluate whether the post-stress LVEF impairment could be related to the post-stress end-systolic ventricular dilation resulting from post ischemic endocardial stunning. Two hundred and eighty-two consecutive patients were studied by conventional diagnostic 2 day stress/rest gated SPECT following injection of 925 MBq of 99mTc-tetrofosmin using a dual-headed SPECT camera. One hundred and forty-seven of these patients (52%) showed reversible perfusion defects, 69 (24%) permanent defects and the remaining 66 (24%) had normal perfusion. One hundred and thirty-eight of these patients had a history of myocardial infarction (MI) and 19% underwent coronary angiography without an intervening cardiac event. Perfusion was analysed on ungated images using 20 segments scored on a five-point scale (0, normal; 4, no uptake), while wall thickening (WT) was assessed visually on stress/rest end-systolic images using a four-point score (0, normal; 3, absence of WT). LVEF and volumes were calculated using an automatic algorithm. The post-stress and rest ratios were determined for both end-diastolic (EDV) and end-systolic (ESV) volume. Normal values for all these parameters were obtained using data from 149 patients with a low likelihood (<5%) of coronary artery disease (CAD). In 50 of the 147 (34%) of patients with reversible perfusion defects, post-stress LVEF was >5% lower than rest values (stunned group), while the remaining 97 patients did not show a significant LVEF change (group 2A). The percentage of patients who developed exercise-induced angina, the percentage of patients who underwent coronary angiography and the segmental summed perfusion and WT scores were significantly higher in the stunned group compared with group 2A. Only ESV increased significantly post-stress, and this increase occurred only in stunned patients. Both EDV and ESV ratios were significantly higher in the stunned group compared with normal controls (P=0.008 and P<0.000001, respectively) and with the subgroup 2A (P=0.011 and P<10(-12), respectively). The ESV stress/rest ratio correlated significantly with the summed WT difference score by univariate analysis in stunned patients. It can be concluded that the post-stress ESV dilation, obtained by stress/rest gated SPECT, seems to be due to endocardial post-ischemic stunning. The stunned patients showed more severe clinical, angiographic, perfusion and function parameters. PMID- 11403182 TI - Localizing infection with a technetium-99m-labeled peptide: initial results. AB - In vitro-labeled leukocyte imaging is useful for the detection of infection, but an in vivo labeling method is preferable. This study sought to evaluate the safety and efficacy of a leukocyte-avid peptide for the detection of infection, to determine the effects of peptide dose on performance and to compare the peptide with in vitro-labeled leukocytes. A 23-amino acid peptide, P483, containing the platelet factor-4 heparin-binding sequence, was labeled with 99mTc and complexed with heparin (P483H). Thirty patients were injected with 29 microg (n = 11), 145 microg (n = 10) or 290 microg (n = 9) of labeled peptide, and imaged 15 min and 90-120 min later. Early and late images were interpreted individually and jointly. Twenty patients underwent (111)In-labeled leukocyte scintigraphy. Fourteen patients had infection: osteomyelitis (n = 7), vascular graft (n = 2), abscess (n = 2), joint replacement (n = 1), surgical wound (n = 1) and pneumonia (n = 1). There were 10 adverse events in six patients; all were mild and resolved spontaneously, and without any intervention. The sensitivity, specificity and accuracy were the same for both early and late imaging: 0.86, 0.81 and 0.83, respectively. Interpreting early and late images together did not improve the results. No relationship between peptide dose and study accuracy was found. In patients undergoing both examinations, the accuracies of the peptide and in vitro-labeled leukocyte imaging were identical: 0.80. In summary, 99mTc P483H safely, rapidly and accurately detected focal infection, was comparable with in vitro-labeled leukocyte imaging and therefore merits further investigation. PMID- 11403183 TI - Quantification of reconstructed SPECT non-uniformity. AB - A simple method is presented for the quantification of reconstructed image non uniformity which is both reproducible and sensitive to changes in system response. Each reconstructed transaxial slice is converted into a polar image by stacking together radial profiles measured from the image centre at 15 degree intervals. Calculating the intensity variations of each polar image in the horizontal and vertical directions then generates measures of radial and tangential uniformity, respectively. Axial uniformity is assessed by stacking together the average radial profiles for each slice and monitoring the variation in intensity of each one from the average profile for the whole phantom. The reproducibility and sensitivity of the indices were assessed using synthetic projection data generated at different count densities and with varying levels of planar non-uniformity. There is little improvement in either the reproducibility or the sensitivity of the indices for count densities in excess of 1 million counts per slice provided that the data have been prefiltered using an appropriate filter. The indices are capable of detecting a range of artefacts relevant to clinical imaging and can be used as part of a quality control programme or to compare the performance of different single photon emission computed tomography (SPECT) systems. PMID- 11403184 TI - 57Co SPECT, 99mTc-ECD SPECT, MRI and neuropsychological testing in senile dementia of the Alzheimer type. AB - Inflammatory mechanisms contribute to the pathophysiology of senile dementia of the Alzheimer type (sDAT). Previous studies have shown that 57Co single photon emission computed tomography (SPECT) is able to visualize inflammatory lesions, probably by means of the final common pathway of Ca2+ homeostasis disturbance in both neuronal degeneration and inflammation. The aims of this study were: (1) to detect 57Co SPECT changes in sDAT patients; (2) to correlate these findings with those of conventional neuroimaging techniques and neuropsychological testing (NPT); and (3) to compare 57Co SPECT findings in sDAT patients with those in other types of dementia. Six patients suffering from probable sDAT were included and compared with four patients suffering from other types of dementia. All patients had a magnetic resonance imaging (MRI) scan, NPT, 57Co and 99mTc-ethyl cysteinate dimer (ECD) SPECT scan. Perfusion SPECT images were semiquantitatively evaluated by comparison with an age-matched normal database, while 57Co SPECT scans were assessed qualitatively. MRI and 99mTc-ECD SPECT scans yielded conclusive results with regard to the exclusion of other pathologies and the confirmation of the diagnosis. Using visual analysis, 57Co SPECT scans were unable to show any regional raised uptake, irrespective of the disorder, depth or extent of the perfusion defects, presence of atrophy on MRI or the results of NPT. PMID- 11403185 TI - Striatal dopamine transporter binding in early to moderately advanced Parkinson's disease: monitoring of disease progression over 2 years. AB - Imaging of striatal dopamine transporter binding allows differentiation between patients with Parkinson's disease and controls. We investigated the use of this technique to monitor disease progression. We used N-(3-iodopropen-2-yl)-2beta carbomethoxy-3beta-(4-chlorophenyl) tropane (IPT) and single photon emission computed tomography (SPECT) to determine dopamine transporter function in eight patients with Parkinson's disease Hoehn and Yahr stage I to III over time. Patients were recruited from the movement disorder clinic and were studied at entry and after a follow-up period of 1 and 2 years. Specific striatal IPT binding was measured with a manual region of interest technique. At entry, all patients showed a reduction of striatal IPT uptake of approximately 50% compared to controls, with a mean striatum to background ratio of 3.61 +/- 0.72 (controls, 7.34 +/- 1.18). Putamen to background ratios were initially measured as 2.42 +/- 0.74 and caudate to background ratios as 5.00 +/- 0.73 (controls, 6.46 +/- 1.22 for putamen and 8.58 +/- 1.36 for caudate). Specific striatal IPT binding decreased by a mean of 6.6% in the first year and another 5.3% in the second year. Changes of specific IPT binding over time were similar in caudate and putamen. In patients with clinically asymmetric disease, differences between the rate of decline in the ipsilateral and contralateral sides could not be detected. The findings suggest that IPT SPECT can quantify the reduction of dopamine transporter binding over time. This technique seems to be a useful tool in monitoring the intra-individual progression of dopaminergic cell loss in patients with Parkinson's disease and may help to follow the effects of putative neuroprotective drugs in future clinical trials. PMID- 11403186 TI - Molecular epidemiology of Neisseria gonorrhoeae exhibiting decreased susceptibility and resistance to ciprofloxacin in Hawaii, 1991-1999. AB - BACKGROUND: Clinically significant resistance to Centers for Disease Control and Prevention (CDC)-recommended doses of fluoroquinolones (ciprofloxacin and ofloxacin) has been reported for Neisseria gonorrhoeae. In Hawaii, fluoroquinolone-resistant gonococcal isolates were first identified in 1991. GOAL: To assess the diversity, based on phenotypic and genotypic characterization, of gonococcal isolates exhibiting decreased susceptibility (CipI; MICs = 0.125-0.5 microg/ml) or clinically significant resistance (CipR; MICs > or = 1 microg/ml) to ciprofloxacin in Hawaii from 1991 through 1999. STUDY DESIGN: Antimicrobial susceptibilities, auxotype/serovar (A/S) class, GyrA/ParC alteration patterns, and plasmid profiles were determined for gonococci isolated in Honolulu from 1991 through 1999 that exhibited intermediate or clinically significant resistance to ciprofloxacin. Strain phenotypes were defined by A/S class, GyrA/ParC alteration pattern, and penicillin-tetracycline resistance phenotype supplemented with plasmid profiles for beta-lactamase-producing isolates. RESULTS: Altogether, 68 isolates exhibiting intermediate or clinically significant resistance to ciprofloxacin belonged to 23 and 19 strain phenotypes, respectively. Among the CipI and CipR isolates, 4 and 13 GyrA/ParC alterations patterns were identified, respectively. The 91,95/Asp-86 alteration pattern occurred most frequently among CipR isolates. Forty-four strain phenotypes were represented by only one isolate. In addition, seven pairs and two clusters of isolates were identified. CONCLUSIONS: From 1991 through 1997, few gonococcal strains exhibiting intermediate or clinically significant resistance to CDC recommended doses of fluoroquinolones were identified from Hawaii. Isolates belonged to a large number of phenotypic and genotypic types, suggesting that most cases were imported, with only a few instances in which isolate pairs indicated that secondary transmission of infections had occurred in Hawaii. Beginning in 1998, the number of CipR isolates increased markedly, and more isolates belonged to fewer phenotypic and genotypic types, suggesting either more frequent importation of fewer strain types or the possibility that the endemic spread of a few strains is beginning to occur. PMID- 11403187 TI - Syphilis among HIV-infected mothers and their infants in Texas from 1988 to 1994. AB - BACKGROUND: Syphilis was investigated in a group of HIV-infected women and their infants. GOAL: To assess syphilis morbidity among HIV-infected women and their infants. Among women with syphilis during pregnancy, the risks for delivering an infant with congenital syphilis were assessed. STUDY DESIGN: Through the Pediatric Spectrum of HIV Disease project, Texas infants born to HIV-infected women were identified. After the infants were matched with their mothers, it was determined which had been reported as syphilis cases. RESULTS: In this study 18% of the HIV-infected mothers were reported as syphilis cases, most during pregnancy. Half of these mothers delivered infants (n = 49) with congenital syphilis. Inadequate prenatal care was the only significant risk for delivering an infant with congenital syphilis. The congenital syphilis rate among Texas infants of HIV-infected mothers was 48.8 per 1,000 live births. CONCLUSION: The congenital syphilis rate among Texas infants born to HIV-infected mothers was almost 50 times that of the general population. PMID- 11403188 TI - Self-collection of vaginal swabs for the detection of Chlamydia, gonorrhea, and trichomoniasis: opportunity to encourage sexually transmitted disease testing among adolescents. AB - BACKGROUND: Many sexually transmitted diseases (STDs) are prevalent among adolescents, yet compliance to undergo STD testing by this population is suboptimal. Efforts to enhance compliance with testing among at-risk youth are needed. GOAL: To determine the feasibility and acceptability of self-collection of vaginal swabs for the detection of Chlamydia trachomatis, Neisseria gonorrhoeae, and Trichomonas vaginalis among high-school students attending a school health clinic. STUDY DESIGN: Enrolled in the study were 228 female students between the ages of 15 and 19 years. Each student self-collected a single vaginal swab that was tested for C trachomatis, N gonorrhoeae, and T vaginalis by polymerase chain reaction amplification. Acceptability of self collection of vaginal swabs was assessed. RESULTS: The prevalence of any STD was 18%. Trichomoniasis, chlamydia, and gonorrhea were diagnosed in 10%, 8%, and 2% of students, respectively. Nearly 13% of females who had never previously had a gynecologic examination tested positive for an STD, and 51% of infected students would not have pursued testing by traditional gynecologic examination if self collection was not offered. Self-collection of vaginal swabs was almost uniformly reported as easy to perform (99%) and preferable to a gynecologic examination (84%). Nearly all (97%) stated that they would undergo testing at frequent intervals if self-testing were available. CONCLUSIONS: Self-collected vaginal swabs for STD testing can be easily implemented in a high-school setting with high acceptability among students, enabling the detection of many STDs that would otherwise remain undetected and untreated. PMID- 11403189 TI - Occurrence of trichomoniasis in women in Denmark, 1967-1997. AB - BACKGROUND: The Neisseria Unit at Statens Serum Institut (SSI), Copenhagen, Denmark administers laboratory surveillance systems for gonorrhea and chlamydia, however, no similar system has been available for trichomoniasis. OBJECTIVES: To describe changes in the occurrence of trichomoniasis in women and in the distribution according to age, and to compare these changes with those observed in women with gonorrhea or chlamydia. STUDY DESIGN: Data on the epidemiology of trichomoniasis were based on the annual laboratory records at the Neisseria Unit at SSI. Data on gonorrhea (1967-1997) and chlamydia (1994-1997) originated from the National Laboratory Surveillance Systems. RESULTS: During the period 1967 to 1997, the percentage of Trichomonas vaginalis-positive specimens showed a constant decrease from 19 to fewer than 2. The annual number of specimens received varied between a maximum of 31,626 in 1975 and a minimum of 3,291 in 1997. The median age of women with trichomoniasis increased from 31 to 39 years as compared with a change from 21 to 24 years in women with gonorrhea (1967-1997) and a median age of 22 years in women with chlamydia (1994-1997). CONCLUSIONS: A significant decline in the occurrence of trichomoniasis in women concurrent with a significant rise in the median age of women with the disorder was observed. During the periods investigated, women with trichomoniasis were significantly older than women with gonorrhea or chlamydia. PMID- 11403190 TI - Fleeting foreskins: the misclassification of male circumcision status. AB - BACKGROUND: Errors in the classification of male circumcision status could bias studies linking infection to lack of circumcision. GOAL: To determine the frequency and factors associated with the reproducibility of reporting circumcision status. STUDY DESIGN: Secondary analysis of data using logistic regression modeling from a multicenter randomized controlled trial was performed. RESULTS: At follow-up assessment, 15.6% of clinician reports on circumcision status disagreed with baseline reports. Disagreement was more common if both clinicians were women than if both were men (odds ratio [OR], 2.8; 95% CI, 1.9 4.1). As compared with whites reported as circumcised (4%, 19/532 visits), the highest disagreement involved uncircumcised Hispanic (OR, 3.3; 95% CI, 1.7-6.3), white (OR, 12.2; 95% CI, 5.8-25.6), or black (OR, 17.1; 95% CI, 10.4-27.9) men. CONCLUSIONS: This is one study among a small number of studies examining the reproducibility of clinician-reported circumcision status by comparing multiple clinical examinations of the same patient. The magnitude of the misclassification discovered could bias results and indicates the need for greater accuracy in reporting circumcision status in future studies. PMID- 11403191 TI - Federally funded sexually transmitted disease programs and managed care: a review of current and planned partnerships. AB - BACKGROUND: The Centers for Disease Control and Prevention requested that sexually transmitted disease (STD) programs report their current activities and plans to collaborate with managed care organizations in their 1999 applications for federal funding. GOAL: To review CDC STD program applications for funding to assess the number of activities between STD programs and managed care organizations. METHODS: Narrative data on managed care topics were abstracted from 59 funding applications (50 states, 7 cites or counties, and 2 US territories), using standard qualitative methods. A coding system was applied to categorize each managed care activity into one of nine categories (interrater reliability, 93%). An expert panel ranked activities by complexity, and these scores were used to develop an overall complexity score for each program. RESULTS: All but 9 of the 59 applicants reported managed care organization activities. Altogether, 208 activities were specifically documented, 45% of which were classified as operational in 1999. The most frequently reported activities involved gathering and giving information and promoting STD care through legislation and state Medicaid activities. CONCLUSIONS: Considerable information transfer and policy action between STD programs and managed care organizations are taking place. Further integration of services and policies should be studied and encouraged to promote the effective treatment of STD. PMID- 11403192 TI - Efficacy and tolerability of topical 1% cidofovir cream for the treatment of external anogenital warts in HIV-infected persons. AB - BACKGROUND: Treatment options for anogenital warts in patients with HIV-1 are unsatisfactory because they fail to eradicate latent human papillomavirus. GOAL: To determine tolerability and efficacy of topical 1% cidofovir cream for the treatment of external anogenital warts in HIV-infected patients. STUDY DESIGN: A randomized, placebo-controlled, single-blind, crossover pilot study of either 1% cidofovir cream or placebo applied once daily 5 days a week for 2 weeks followed by 2 weeks of observation was performed. RESULTS: Six patients were randomized to 1% cidofovir cream and six to placebo. The latter patients eventually received 1% cidofovir cream. Thus, 12 treatment rounds of cidofovir were compared with six rounds of placebo. A reduction of more than 50% in the total wart area achieved by seven cidofovir treatments (58%), as compared with no placebo regimen (P = 0.02). Local reactions occurred in 10 of the 12 patients treated with cidofovir, as compared with 0 of the 6 subjects in the placebo group (P < 0.001). CONCLUSIONS: For the initial clearance of anogenital warts in HIV-infected patients, 1% cidofovir cream is significantly more effective than vehicle cream. Local mucosal erosion is a common side effect. PMID- 11403193 TI - Effect of Chlamydia trachomatis coinfection on HIV shedding in genital tract secretions. PMID- 11403194 TI - Prevention of sexually transmitted diseases and HIV in young military men: evaluation of a cognitive-behavioral skills-building intervention. AB - BACKGROUND: Military personnel deployed to foreign countries with high endemic rates of sexually transmitted diseases (STDs) are at risk. GOAL: To evaluate a cognitive-behavioral, skills-building intervention to prevent STDs in junior, enlisted, male US Marines deployed to the Western Pacific aboard ships with periodic liberty visits to foreign ports. STUDY DESIGN: This study, using a quasi experimental design, assigned 584 men to a cardiopulmonary resuscitation training (n = 288) or a cognitive-behavioral intervention (n = 296). The intervention aimed to increase prevention knowledge, reduce high-risk psychosocial (motivation) factors, and build decision-making and communication skills to reduce risky sexual behaviors and alcohol consumption. RESULTS: Participation in the intervention was associated with increased knowledge regarding symptoms and treatment of STDs and HIV (P < 0.001), decreased alcohol use (P < 0.01), and sexual risk (P < 0.01) during liberty ports of call. CONCLUSION: A multiple session, cognitive-behavioral, skills-building intervention can be successfully implemented for deployed military personnel. PMID- 11403195 TI - Chlamydia, gonorrhea, and HIV-1 prevalence among five populations of women in the Czech and Slovak Republics. AB - BACKGROUND: Five populations at risk for sexually transmitted diseases (STDs) in the Czech and Slovak Republics were sampled. GOAL: To estimate prevalence of Chlamydia trachomatis, Neisseria gonorrhoeae, and HIV-1 infections. STUDY DESIGN: Urine specimens were collected serially from women at a Prague prenatal clinic (n = 134), a Prague dermatovenerealogy clinic (n = 91), sex workers from northern and central Bohemia (n = 35), students from a northern Bohemian school (n = 217), and Gypsies from Jarovnice, Slovakia (n = 128). These specimens were tested for chlamydia and gonorrhea using a ligase chain reaction pooling algorithm, and for HIV using an enzyme immunoassay confirmed by Western blot. RESULTS: The prevalence of chlamydia was 2.2% (95% CI, 0.4-6.4) in the prenatal clinic, 5.5% (95% CI, 1.8-12.4) in the STD clinic, 22.9% (95% CI, 10.4-40.1) among street sex workers, 8.2% (95% CI, 3.6-15.6) among sexually active female high school students, and 3.9% (95% CI, 1.3-8.9) among Gypsy women. Gonorrhea was found in only two populations: 2.2% (95% CI, 0.3-7.7) in the STD clinic, and 2.9% (95% CI, 0.1-14.9) among sex workers. No HIV-1 infection was detected. CONCLUSIONS: Urine screening was an efficient and accurate method for identifying groups at risk for STDs in the Czech Republic and Slovakia because sample collection was fast and noninvasive, and potential participation bias was reduced by high acceptability. PMID- 11403196 TI - Maximum-likelihood expectation-maximization reconstruction of sinograms with arbitrary noise distribution using NEC-transformations. AB - The maximum-likelihood (ML) expectation-maximization (EM) [ML-EM] algorithm is being widely used for image reconstruction in positron emission tomography. The algorithm is strictly valid if the data are Poisson distributed. However, it is also often applied to processed sinograms that do not meet this requirement. This may sometimes lead to suboptimal results: streak artifacts appear and the algorithm converges toward a lower likelihood value. As a remedy, we propose two simple pixel-by-pixel methods [noise equivalent counts (NEC)-scaling and NEC shifting] in order to transform arbitrary sinogram noise into noise which is approximately Poisson distributed (the first and second moments of the distribution match those of the Poisson distribution). The convergence speed associated with both transformation methods is compared, and the NEC-scaling method is validated with both simulations and clinical data. These new methods extend the ML-EM algorithm to a general purpose nonnegative reconstruction algorithm. PMID- 11403197 TI - Penalized discriminant analysis of [15O]-water PET brain images with prediction error selection of smoothness and regularization hyperparameters. AB - We propose a flexible, comprehensive approach for analysis of [15O]-water positron emission tomography (PET) brain images using a penalized version of linear discriminant analysis (PDA). We applied it to scans from 20 subjects (eight scans/subject) performing a finger movement task and analyzed: 1) two classes to obtain a covariance-normalized baseline-activation image, and 2) eight classes for the mean within subject temporal structure which contained baseline activation and time-dependent changes in a two-dimensional canonical subspace. We imposed spatial smoothness on the resulting image(s) by expanding it in five tensor-product B-spline (TPS) bases of varying smoothness, and further regularized with a ridge-type penalty on the noise covariance matrix. The discrimination approach of PDA provides a probabilistic framework within which prediction error (PE) estimates are derived. We used these to optimize over TPS bases and a ridge hyperparameter (expressed as equivalent degrees of freedom, EDF). We obtained unbiased, low variance PE estimates using modern resampling tools (.632+ Bootstrap and cross validation), and compared PDA of 1) TPS projected, mean-normalized and unnormalized scans and 2) mean-normalized scans with and without additional presmoothing. By examining the tradeoffs between PE and EDF, as a function of basis selection and image smoothing we demonstrate the utility of PDA, the PE framework, and the relationship between singular value decomposition and smooth TPS bases in the analysis of functional neuroimages. PMID- 11403198 TI - Hierarchical estimation of a dense deformation field for 3-D robust registration. AB - A new method for medical image registration is formulated as a minimization problem involving robust estimators. We propose an efficient hierarchical optimization framework which is both multiresolution and multigrid. An anatomical segmentation of the cortex is introduced in the adaptive partitioning of the volume on which the multigrid minimization is based. This allows to limit the estimation to the areas of interest, to accelerate the algorithm, and to refine the estimation in specified areas. At each stage of the hierarchical estimation, we refine current estimate by seeking a piecewise affine model for the incremental deformation field. The performance of this method is numerically evaluated on simulated data and its benefits and robustness are shown on a database of 18 magnetic resonance imaging scans of the head. PMID- 11403199 TI - Contextual clustering for analysis of functional MRI data. AB - We present a contextual clustering procedure for statistical parametric maps (SPM) calculated from time varying three-dimensional images. The algorithm can be used for the detection of neural activations from functional magnetic resonance images (fMRI). An important characteristic of SPM is that the intensity distribution of background (nonactive area) is known whereas the distributions of activation areas are not. The developed contextual clustering algorithm divides an SPM into background and activation areas so that the probability of detecting false activations by chance is controlled, i.e., hypothesis testing is performed. Unlike the much used voxel-by-voxel testing, neighborhood information is utilized, an important difference. This is achieved by using a Markov random field prior and iterated conditional modes (ICM) algorithm. However, unlike in the conventional use of ICM algorithm, the classification is based only on the distribution of background. The results from our simulations and human fMRI experiments using visual stimulation demonstrate that a better sensitivity is achieved with a given specificity in comparison to the voxel-by-voxel thresholding technique. The algorithm is computationally efficient and can be used to detect and delineate objects from a noisy background in other applications. PMID- 11403200 TI - Multistage hybrid active appearance model matching: segmentation of left and right ventricles in cardiac MR images. AB - A fully automated approach to segmentation of the left and right cardiac ventricles from magnetic resonance (MR) images is reported. A novel multistage hybrid appearance model methodology is presented in which a hybrid active shape model/active appearance model (AAM) stage helps avoid local minima of the matching function. This yields an overall more favorable matching result. An automated initialization method is introduced making the approach fully automated. Our method was trained in a set of 102 MR images and tested in a separate set of 60 images. In all testing cases, the matching resulted in a visually plausible and accurate mapping of the model to the image data. Average signed border positioning errors did not exceed 0.3 mm in any of the three determined contours-left-ventricular (LV) epicardium, LV and right-ventricular (RV) endocardium. The area measurements derived from the three contours correlated well with the independent standard (r = 0.96, 0.96, 0.90), with slopes and intercepts of the regression lines close to one and zero, respectively. Testing the reproducibility of the method demonstrated an unbiased performance with small range of error as assessed via Bland-Altman statistic. In direct border positioning error comparison, the multistage method significantly outperformed the conventional AAM (p < 0.001). The developed method promises to facilitate fully automated quantitative analysis of LV and RV morphology and function in clinical setting. PMID- 11403201 TI - Three-dimensional texture analysis of MRI brain datasets. AB - A method is proposed for three-dimensional (3-D) texture analysis of magnetic resonance imaging brain datasets. It is based on extended, multisort co occurrence matrices that employ intensity, gradient and anisotropy image features in a uniform way. Basic properties of matrices as well as their sensitivity and dependence on spatial image scaling are evaluated. The ability of the suggested 3 D texture descriptors is demonstrated on nontrivial classification tasks for pathologic findings in brain datasets. PMID- 11403202 TI - Automated CT image evaluation of the lung: a morphology-based concept. AB - Computed tomography (CT) provides the most reliable method to detect emphysema in vivo. Commonly used methods only calculate the area of low attenuation [pixel index (PI)], while a radiologist considers the bullous morphology of emphysema. The PI is a good, well-known measure of emphysema. But it is not able to detect emphysema in cases in which emphysema and fibrosis occur at the same time. This is because fibrosis leads to a low number of low-attenuation pixels, while emphysema leads to a high number of pixels. The PI takes the average of both and, consequently, may present a result within the normal range. METHOD: The main focus of this paper is to present a new algorithm of thoracic CT image evaluation based on pulmonary morphology of emphysema. The PI is extended, in that it is enabled to differentiate between small, medium, and large bullae (continuous low attenuation areas). It is not a texture-based algorithm. The bullae are sorted by size into four size classes: class 1 being within the typical size of lung parenchyma; classes 2-4 presenting small, medium, and large bullae. It is calculated how much area the different classes take up of all low-attenuation pixels. The bullae index (BI) is derived from the percentage of areas covered, respectively, by small, medium, and large bullae. From the relation of the area of bullae belonging to class 4, to that of those belonging to class 2, a measure of the emphysema type (ET)is calculated. It classifies the lung by the type of emphysema in bullous emphysema or small-sized, diffuse emphysema, respectively. RESULTS: The BI is as reliable as the PI. In cases in which the PI indicates normal values while in fact emphysema is coexisting with fibrosis, the BI, nevertheless, detects the destruction caused by the emphysema. The BI combined with the ET reflects the visual assessment of the radiological expert. CONCLUSION: The BI is an objective and reliable index in order to quantify emphysematous destruction, hence, avoiding interobserver variance. This is particularly interesting for follow-up. The classification of the ET is a helpful and unique approach to achieving an exact diagnosis of emphysema. PMID- 11403203 TI - Fractal analysis of bone X-ray tomographic microscopy projections. AB - Fractal analysis of bone X-ray images has received much interest recently for the diagnosis of bone disease. In this paper, we propose a fractal analysis of bone X ray tomographic microscopy (XTM) projections. The aim of the study is to establish whether or not there is a correlation between three-dimensional (3-D) trabecular changes and two-dimensional (2-D) fractal descriptors. Using a highly collimated beam, 3-D bone X-ray tomographic images were obtained. Trabecular bone loss was simulated using a mathematical morphology method. Then, 2-D projections were generated in each of the three orthogonal directions. Finally, the model of fractional Brownian motion (fBm) was used on bone XTM 2-D projections to characterize changes in bone structure that occur during disease, such a simulation of bone loss. Results indicate that fBm is a robust texture model allowing quantification of simulations of trabecular bone changes. PMID- 11403204 TI - Hypothermia attenuates beta1 integrin expression on extravasated neutrophils in an animal model of meningitis. AB - Brain injury in meningitis occurs in part as a consequence of leukocyte migration and activation. Leukocyte integrins are pivotal in the inflammatory response by mediating adhesion to vascular endothelium and extracellular matrix proteins. We have demonstrated that moderate hypothermia early in the course of meningitis decreases leukocyte sequestration within the brain parenchyma. This study examines whether hypothermia alters neutrophil integrin expression in a rabbit model of bacterial meningitis. Prior to the induction of meningitis, peripheral blood samples were obtained and the neutrophils isolated. Sixteen hours after inducing group B streptococcal meningitis, animals were treated with antibiotics, i.v. fluids, and mechanically ventilated. Animals were randomized to hypothermia (32-33 degrees C) or normothermia conditions. After 10 hours of hypothermia or normothermia, neutrophils were isolated from the blood and cerebral spinal fluid (CSF), stained for beta1 and beta2 integrins, and analyzed using flow cytometry. Cerebral spinal fluid neutrophil beta1 integrin expression was significantly decreased in hypothermic animals. Beta-1 integrins can assume a higher affinity or "activated" state following inflammatory stimulation. Expression of "activated" beta1 integrins was also significantly decreased in hypothermic animals. Beta2 CSF neutrophil integrin expression was decreased in hypothermic animals, but failed to reach significance. These data suggest hypothermia may attenuate extravasated leukocyte expression of both total and "activated" beta1 integrins. PMID- 11403205 TI - Dysregulated cytokine production in human cystic fibrosis bronchial epithelial cells. AB - Although pulmonary inflammation is an important pathologic event in cystic fibrosis (CF), the relationship between expression of the CF gene and the inflammatory response is unclear. We studied tumor necrosis factor (TNF) alpha and IL-1beta stimulated production of IL-6 and IL-8 by CF, corrected CF, and normal human bronchial epithelial cells in culture. During the first 24 hours of TNFalpha stimulation, CF cells produced significantly more IL-8 than normal or corrected CF cells. In the second 24 hours of TNFalpha stimulation, IL-6 and IL-8 generation ceased in normal and corrected CF cells but accelerated in CF cells, resulting in marked IL-6 and IL-8 accumulation in CF cells. Similar results were found when cells were stimulated with IL-1beta. Finally, when CF cells were grown at 27 degrees C (a culture condition which results in transport of CF transmembrane conductance regulator, CFTR, to the cell membrane and normalization of chloride conductance) TNFalpha-stimulated production of IL-6 and IL-8 reverted to normal. We conclude that dysregulation of cytokine generation by CF bronchial epithelial cells is directly related to expression of mutant CFTR and these observations provide a potential mechanism for persistence of airway inflammation in CF. PMID- 11403206 TI - Poly ADP ribose-polymerase inhibitors prevent the upregulation of ICAM-1 and E selectin in response to Th1 cytokine stimulation. AB - We studied the role of poly-ADP-ribose polymerase (PARP) in the mobilization of ICAM-1, VCAM-1, and E-selectin by TNF-alpha and IL-1beta in cultured human endothelial cells. Enzyme linked immunosorbent analysis (ELISA) was used to assess if ICAM-1, VCAM-1, and E-selectin were expressed at the cell surface, and if PARP inhibition (using the selective PARP inhibitor GPI 6150) blocked the induced expression. Endothelial cell adhesion molecule expression was evaluated at 4 and at 24 h after cytokine stimulation. At 4 h ICAM-1 and E-selectin, but not VACM-1, were stimulated by both IL-1beta and TNF-alpha. Blocking PARP via GPI 6150 only affected TNF-alpha induced E-selectin expression at 4 hours. ICAM-1, VCAM-1, and E-selectin expression were all stimulated by both IL-1beta and TNF alpha in the 24 h assays. PARP inhibition with GPI 6150 blocked the IL-1beta mediated stimulation of both ICAM-1 and E-selectin expression, and blocked TNF alpha stimulation of ICAM-1 expression at 24 h. These experiments suggest that specific PARP inhibition may provide a novel method of controlling leukocyte dependent inflammation through the reduction of ICAM-1 and E-selectin expression in endothelial cells in response to cytokines. PMID- 11403207 TI - An intact cytoskeleton is required for prolonged respiratory burst activity during neutrophil phagocytosis. AB - The temporal relationship between phagocytosis and respiratory burst activity was investigated. Neutrophil uptake of yeast particles was synchronized and the kinetics of the oxidative burst was determined using an isoluminol/luminol amplified chemiluminescence system. The reactive oxygen species were mainly generated intracellularly (defined as the activity that remained in an luminol enhanced system in the presence of superoxide dismutase and catalase). Following phagocytosis, the intracellular response rapidly reached a level close to the maximum and the activity was almost constant for the first 10 to 15 min. The response then slowly declined. The presence of cytochalasin B, an inhibitor of actin polymerization, greatly reduced the respiratory burst activity, and this was true also when the inhibitor was added after completion of uptake of yeast particles. Our results thus show that there is a continuous production of oxygen metabolites long after phagocytosis is completed. There is also a requirement for an intact cytoskeleton for prolonged superoxide production inside the phagosome. PMID- 11403208 TI - Species dependence for binding of small molecule agonist and antagonists to the C5a receptor on polymorphonuclear leukocytes. AB - This study investigated the receptor binding affinities of a C5a agonist and cyclic antagonists for polymorphonuclear leukocytes (PMNs) isolated from human, sheep, pig, dog, rabbit, guinea pig, rat and mouse. The affinities of the two small molecule antagonists, F-[OPdChaWR] and AcF-[OPdChaWR], and the agonist, YSFKPMPLaR, revealed large differences in C5a receptor (C5aR) affinities between species. The antagonists bound to human, rat and dog PMNs with similar high affinities, but with lower affinities to PMNs from all other species. The C5a agonist also bound with varying affinities between species, but showed a different affinity profile to the antagonists. In contrast, recombinant human C5a had similar affinity for PMNs of all species investigated. The low correlation between the affinities of the antagonists and the agonist between species either suggests that different receptor residues are important for distinguishing between agonist/antagonist binding, or that the agonist and antagonist peptides bind to two distinct sites within the C5aR. PMID- 11403209 TI - Homocysteine augments cytokine-induced chemokine expression in human vascular smooth muscle cells: implications for atherogenesis. AB - Hyperhomocysteinemia is an independent risk factor for atherosclerosis and atherothrombosis. While in vitro studies have revealed a number of homocysteine mediated alterations in the thromboregulatory properties of endothelial cells, comparatively little is known about homocysteine-modulated smooth muscle cell function. We observed that exposure of human aortic smooth muscle cells to pathophysiologically relevant concentrations of homocysteine results in concentration-dependent increases in cytokine-induced MCP-1 and IL-8 secretion. RNase protection assays revealed that both MCP-1 and IL-8 mRNA concentrations are increased in homocysteine-treated smooth muscle cells when compared to cells activated with cytokines alone. Homocysteine treatment also increased cytosolic to-nuclear translocation of the p65 and p50 subunits of the Rel/NF-kappaB family of transcription factors but had no effect on AP-1 activation. Cumulatively, these data suggest that homocysteine may increase monocyte recruitment into developing atherosclerotic lesions by upregulating MCP-1 and IL-8 expression in vascular smooth muscle cells. PMID- 11403210 TI - Effects of heat pretreatment on histopathology, cytokine production, and surfactant in endotoxin-induced acute lung injury. AB - To determine the effect of heat stress on histopathology of acute lung injury (ALI) caused by administration of lipopolysaccharide (LPS), and to determine the roles of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, interferon (IFN)-gamma, IL-10 and surfactants in heat-induced tolerance to ALI, we administered either saline or LPS (3 mg/kg of body weight) intravenously to male Sprague-Dawley rats without and with heat pretreatment. Five hours after LPS or saline treatment (23 h after heat-pretreatment), samples were obtained. We found that the histopathologic features of LPS-induced ALI were attenuated by heat pretreatment. Heat-pretreatment did not decrease the elevated plasma or BAL fluid levels of TNF-alpha, IL-1beta, and IFN-gamma by LPS. The plasma level of IL-10 in LPS-treated rats with heat-pretreatment, however, was increased compared to that of LPS-treated rats without heat-pretreatment (P = 0.001). There were no differences in the BAL fluid concentrations of light or heavy density pulmonary surfactant phospholipids depending on heat-pretreatment in LPS-treated rats. These observations suggest that IL-10 might play a role in decreasing LPS-induced acute lung injury after heat-pretreatment. PMID- 11403211 TI - Adrenomedullin suppresses fMLP-induced upregulation of CD11b of human neutrophils. AB - In this study we investigated the effect of adrenomedullin (AM) on fMLP-mediated activation of human neutrophils. AM partially, but significantly, suppressed fMLP induced upregulation of CD11b expression. The inhibitory effects of AM upon fMLP induced upregulation of CD11b expression were completely blocked by CGRP [8-37], a CGRP receptor antagonist. AM significantly increased cAMP content in neutrophils and SQ-22,536, an adenylate cyclase inhibitor, and KT-5720, a PKA inhibitor, significantly blocked the inhibitory effects of AM upon fMLP-induced upregulation of CD11b expression. This study indicates that binding of AM to the CGRP receptor suppresses fMLP-induced upregulation of CD11b expression of human neutrophils by increasing intracellular cAMP levels. AM may play an important role in the regulation of inflammatory processes, especially in the binding of neutrophils to vascular endothelial cells and subsequent neutrophil emigration evident in acute pulmonary inflammation. PMID- 11403212 TI - Inhibition of the adherence of T-lymphocytes to epithelial cells by a cyclic peptide derived from inserted domain of lymphocyte function-associated antigen-1. AB - Tissue inflammation is characterized by aggravated leukocyte infiltration into the sites of inflammation. The mechanism requires the interactions of leukocyte adhesion-molecules and their ligands in the inflamed tissues. In this study, we demonstrate that a cyclic peptide cLAB.L [cyclol, 12-PenlTDGEATDSGC], derived from the "inserted" or I-domain of LFA-1 is able to inhibit the adherence of T lymphocytes to the epithelial cell monolayers. This inhibition has been thought to involve the disruption of LFA-1/ICAM-1 interaction. The heterotypic adhesion of phorbol ester-activated Molt-3 cells and IFN-gamma-induced Caco-2 monolayers was inhibited upon treatment of the monolayers with monoclonal antibodies (MAbs) to adhesion molecules or with cLAB.L peptide. The adhesion can be inhibited by MAbs to ICAM-1, ICAM-2, and VCAM-1, and cLAB.L peptide in a concentration dependent manner. However, none of the individual uses of these molecules led to a total inhibition. The inhibitory activity of cLAB.L is greatly reduced by low temperature and the absence of cell activation. Treatment of cLAB.L peptide may trigger an early event of apoptosis on activated but not on non-activated Molt-3 cells; no indication of peptide-induced apoptosis was found on Caco-2 cells. Taken together, data from this work suggest that cLAB.L may have applications to direct cell-targeted delivery during tissue inflammation. PMID- 11403213 TI - Are the salutogenic effects of social supports modified by income? A test of an "added value hypothesis". AB - Older adults (54 men, 113 women; M age = 69.5 years) were examined to test the hypothesis that social supports would be more salutogenic (health promoting) for persons with lower incomes than for persons with higher incomes. Interactions of income and social supports (mean of 3 emotional scales of the Interpersonal Support Evaluation List) at study entry predicted changes 15-18 months later in a cardiovascular composite (linear combination of high-density lipoproteins-mean arterial pressure; p < .05), and natural killer cell activity (p < .05). For both outcomes, emotional supports were salutogenic for persons with lower incomes (< or =$29,000/year), but not for persons with higher incomes (>$29,000/year). In contrast, interactions of the Tangible Support Scale with income did not occur. Persons with lower incomes may derive benefits from social supports that go beyond tangible assistance. PMID- 11403214 TI - Projection in surrogate decisions about life-sustaining medical treatments. AB - To honor the wishes of an incapacitated patient, surrogate decision makers must predict the treatment decisions patients would make for themselves if able. Social psychological research, however, suggests that surrogates' own treatment preferences may influence their predictions of others' preferences. In 2 studies (1 involving 60 college student surrogates and a parent, the other involving 361 elderly outpatients and their chosen surrogate decision maker), surrogates predicted whether a close other would want life-sustaining treatment in hypothetical end-of-life scenarios and stated their own treatment preferences in the same scenarios. Surrogate predictions more closely resembled surrogates' own treatment wishes than they did the wishes of the individual they were trying to predict. Although the majority of prediction errors reflected inaccurate use of surrogates' own treatment preferences, projection was also found to result in accurate prediction more often than counterprojective predictions. The rationality and accuracy of projection in surrogate decision making is discussed. PMID- 11403215 TI - Posttraumatic growth following breast cancer: a controlled comparison study. AB - Cancer may be viewed as a psychosocial transition with the potential for positive and negative outcomes. This cross-sectional study (a) compared breast cancer (BC) survivors' (n = 70) self-reports of depression, well-being, and posttraumatic growth with those of age- and education-matched healthy comparison women (n = 70) and (b) identified correlates of posttraumatic growth among BC survivors. Groups did not differ in depression or well-being, but the BC group showed a pattern of greater posttraumatic growth, particularly in relating to others, appreciation of life, and spiritual change. BC participants' posttraumatic growth was unrelated to distress or well-being but was positively associated with perceived life threat, prior talking about breast cancer, income, and time since diagnosis. Research that has focused solely on detection of distress and its correlates may paint an incomplete and potentially misleading picture of adjustment to cancer. PMID- 11403216 TI - Sex differences in predictors of adolescent smoking cessation. AB - Sex differences in predictors of smoking cessation were investigated among 337 male and 490 female participants in the RAND adolescent panel study. Participants reported smoking at least 11-20 times during the past year at Grade 10, with cessation defined as not smoking during the past year at Grade 12. Controlling for demographics, sex-specific analyses indicated that girls who quit smoking within 2 years had friends who smoked less frequently, perceived less parental approval of their smoking, had weaker intentions to continue smoking, used marijuana less frequently, attended fewer different schools, were more likely to have an intact nuclear family, experienced greater peer support, and rated themselves as healthier. Similar analyses for boys yielded results that were generally weaker and nonsignificant, with smoking quantity accounting for several associations in the sex-specific models. Despite these differences, interaction tests revealed significant sex differences for only three predictors. Implications of these results for understanding adolescent smoking cessation are discussed. PMID- 11403217 TI - The influence of school environment and self-regulation on transitions between stages of cigarette smoking: a multilevel analysis. AB - In this research, hierarchical linear modeling (HLM) was used to address how school context influences the likelihood of transitioning between stages of cigarette smoking as well as modifies the individual-level risk factor of self regulation. Survey data were collected from 25,186 middle and high school students attending 38 public schools in Kentucky. Results show that students are less likely to increase use in schools with higher levels of teacher discipline and faculty involvement. The analyses of the multi-level interactions between self-regulation and school context reveal that students possessing low emotional regulation are more likely to initiate experimental smoking in schools with poor levels of discipline and involvement than similar types of students in schools with higher levels of these characteristics. This study illustrates how psychological risk factors for substance use may vary across social environments. PMID- 11403218 TI - Coping, control, and adjustment in Type 2 diabetes. AB - The relationships of both coping strategies and perceived control to psychological and physiological adjustment were investigated in 115 adults (65 women, 50 men) with Type 2 diabetes. Results showed that (a) emotional preoccupation and palliative coping were positively correlated with depression and state anxiety, whereas perceived control was negatively correlated with depression, state anxiety, and hemoglobin A1c (HbA1c); (b) instrumental coping predicted lower depression; (c) perceived control moderated the relationships between instrumental coping and depression, and emotional preoccupation coping and HbA1c; and (d) emotional preoccupation coping mediated the relationships between perceived control and depression, and perceived control and state anxiety. Results are discussed in terms of the goodness-of-fit hypothesis (V. J. Conway & D. J. Terry, 1992), optimal coping, and the importance of perceived control in psychological and physiological adjustment. PMID- 11403219 TI - Effects of coping style and BRCA1 and BRCA2 test results on anxiety among women participating in genetic counseling and testing for breast and ovarian cancer risk. AB - Using the monitoring process model (MPM), the authors examined the immediate effects of coping style and test results on the psychological distress of women at increased risk for breast and/or ovarian cancers. Cases selected for analysis were 107 probands and relatives of positive probands participating in genetic counseling and testing for heritable cancer risk. Specifically, the authors explored the relationships among coping style (high and low monitoring), test results (BRCA1 and BRCA2 mutation carrier and noncarrier status), and psychological distress (state anxiety). Consistent with the MPM, higher monitoring was associated with greater psychological distress while anticipating genetic test results. After test results were disclosed, greater distress was associated with testing positive for a mutation. The implications of the findings for breast and ovarian cancer patients are discussed. PMID- 11403220 TI - Depressive symptoms, social support, and personal health behaviors in young men and women. AB - This study investigated the relationship of depressive symptoms, social support, and a range of personal health behaviors in 2,091 male and 3,438 female university students from 16 countries. Depressive symptoms and social support were measured using the short Beck Depression Inventory and the Social Support Questionnaire; 9 personal health behaviors were also assessed. After the authors took age, social support, and clustering by country into account, depressive symptoms were significantly associated with lack of physical activity, not eating breakfast, irregular sleep hours, and not using a seat belt in both men and women, and additionally with smoking, not eating fruit, and not using sunscreen among women. Low social support was independently associated with low alcohol consumption, lack of physical activity, irregular sleep hours, and not using a seat belt in men and women. Bidirectional causal pathways are likely to link health behaviors with depressed mood. PMID- 11403221 TI - Religion and health. PMID- 11403222 TI - Immune responses against the myelin/oligodendrocyte glycoprotein in experimental autoimmune demyelination. AB - Myelin/oligodendrocyte glycoprotein (MOG) is a surface-exposed antigen of myelin and an important target for autoimmune responses which mediate inflammatory demyelination in the central nervous system. Experimentally, MOG induces strong pathogenic T cell responses in many strains of laboratory animals. Immunological studies in humans also identify MOG as a surprisingly prevalent antigenic molecule among the myelin proteins. In addition, the encephalitogenic properties of MOG are linked to the induction of antibody responses which have been demonstrated to directly promote central nervous system demyelination, a hallmark neuropathological feature in disorders such as human multiple sclerosis. Factors responsible for autoimmunity to MOG likely include genetic influences as well as other mechanisms, which are the subject of intense investigation. This article reviews experimental data currently available on specificity and pathogenic roles of T cell and antibody responses against MOG, which have implications relevant to multiple sclerosis and related disorders. PMID- 11403223 TI - Human mast cell apoptosis is regulated through Bcl-2 and Bcl-XL. AB - It is well established that human mast cell proliferation and maturation are regulated by kit ligand (stem cell factor). Little is known, however, about how these two processes are negatively regulated and thus, how mast cell number is controlled in normal and pathologic conditions. We therefore first hypothesized that SCF-dependent human mast cells would undergo programmed cell death (apoptosis) on removal of SCF as has been shown for growth factor-dependent rodent mast cells. We then examined whether SCF acts as a survival factor through the regulation of the bcl-2 family of apoptosis-regulatory genes. As hypothesized, elimination of SCF from primary peripheral blood-derived human mast cell cultures resulted in a significant apoptotic process. During apoptosis, down regulation of the two apoptosis-regulatory proteins Bcl-2 and Bcl-XL was observed. Moreover, a deregulated expression of these two proteins was found in two human mast cell lines which are SCF-independent. Thus, SCF functions as a survival factor by repressing apoptosis of human mast cells through Bcl-2 and Bcl XL. Deregulated expression of these antiapoptotic proteins may contribute to proliferation and accumulation of mast cells in certain forms of systemic mast cell disorders. PMID- 11403224 TI - Systemic administration of immunostimulatory DNA sequences mediates reversible inhibition of Th2 responses in a mouse model of asthma. AB - This study investigated whether immunostimulatory DNA sequences (ISS) induce a transient or sustained inhibition of Th2 responses to inhaled antigen. We sensitized mice with subcutaneous injections to develop a Th2 response to ovalbumin (ova) and then administered a dose of ISS prior to ova inhalation challenge. Mice were then rechallenged with ova by inhalation a second time at varying time points after the first ova inhalation (1 to 8 weeks later) to determine whether the ISS dose administered prior to the first ova inhalation protected against a subsequent second ova inhalation challenge. A single dose of ISS inhibited the Th2 response to the first inhalation of ova antigen, as well as 4 weeks later to the second inhalation of ova. However, ISS did not inhibit a Th2 response to the second inhalation of ova 8 weeks later. The reversible inhibition of Th2 responses at 8 weeks suggests the need for repeated ISS administration at monthly intervals. PMID- 11403225 TI - The regulation of FasL expression--a distinquishing feature between monocytes and T lymphocytes/NK cells with possible implications for SLE. AB - Monocytes and lymphocytes from patients with systemic lupus erythematosus (SLE) had a higher cell surface expression of FasL than the corresponding cells from healthy individuals. Inhibitors of metalloproteases upregulated the surface expression of FasL in peripheral blood lymphocytes (PBL), indicating that a metalloprotease is responsible for the cleavage of FasL. The level of sFasL in serum was slightly increased in the patient group compared to the controls. Therefore, the possible contribution of various mononuclear cell types to the release of FasL was analyzed. Isolated NK cells and T lymphocytes released FasL into the medium and the release was prevented by inhibitors of metalloproteases. In contrast, isolated monocytes did not release FasL. FasR expression was elevated in patients with inverted CD4/CD8 ratio, while FasL expression showed no relationship to CD4/CD8 ratio. The absence of FasL release by isolated cells and a high level of surface expression of FasL distinguish monocytes and T lymphocytes/NK cells. PMID- 11403226 TI - Cytokine modulation with immune gamma-globulin in peripheral blood of normal children and its implications in Kawasaki disease treatment. AB - Intravenous immune gamma-globulin (IVIG) is used successfully in the treatment of Kawasaki disease, with dose-dependent rapid resolution of symptoms such as fever and irritability and a decrease in ESR, WBCs, and platelets. The mode of action of IVIG in reducing this inflammatory response is not clearly understood. Recently anticytokine antibodies in IVIG have been demonstrated. Serum levels of proinflammatory cytokines have been shown to be elevated in patients with Kawasaki disease. The cytokine interleukin-6 (IL-6) is involved in the de novo production of acute-phase proteins by hepatocytes and cause thrombocytosis and fever in response to tissue injury. Patients receiving parenteral recombinant human IL-6 have dose-dependently experienced fever, malaise, chills, and acute phase reaction. With high IL-6 concentrations, central nervous system toxicity has also been reported and IL-6 has been thought to mediate endothelial damage. We evaluated the response of stimulated blood cells of 12 normal children to IVIG in the release of the cytokines IL-6, IL-8, TNF-alpha. and IL-6 receptor (sIL 6R). The levels of cytokines IL-6, IL-8, and TNF-alpha (but not sIL-6R) in peripheral blood induced by stimulation with LPS were markedly reduced (P < 0.008) within 3 hr when incubated with IVIG compared to without IVIG. Thus we demonstrated that cells of normal children respond to IVIG in vitro by reducing cytokines such as IL-8, TNF-alpha, and IL-6 without affecting the level of receptor sIL-6R during an acute inflammatory response. We also found significantly higher IL-6 levels in children with Kawasaki disease compared to children with blood culture-negative febrile illnesses. In five children with Kawasaki disease we measured serum IL-6 before and after IVIG and assessed the clinical response to IVIG therapy. Therapy with IVIG was followed by a rapid resolution of symptoms in Kawasaki disease, with a significant decrease in serum IL-6. The attenuation of proinflammatory cytokine responses, especially IL-6, following infusions of IVIG may play an integral role in the rapid resolution of symptoms and decrease in the acute-phase proteins in children with Kawasaki disease. Cells of normal children were found to respond to the IVIG in a manner similar to that of the Kawasaki children. PMID- 11403227 TI - Interferon beta1a treatment modulates TH1 expression in gammadelta + T cells from relapsing-remitting multiple sclerosis patients. AB - A paradigm exists that multiple sclerosis is causally related to dysregulation of TH1 inflammatory cytokines and TH2 antiinflammatory cytokines. The cytokine source(s) that initiate the imbalances are unknown. In this study, gammadelta, CD4, and CD8 T cell receptor-positive (TCR+) cells were isolated from the blood of 26 definitive relapsing-remitting multiple sclerosis patients prior to interferon beta-1a (IFNbeta1a) therapy and following 8-10 weeks of this therapy. The bioactivities of interferon gamma (IFNgamma), interleukin 10 (IL10), and interleukin 12 (IL12) were determined. The concentrations of IFN-gamma, IL10, and IL12 from each cell type did not change significantly with IFNbeta1a treatment. The IL10 secreted by gammadelta TCR+ cells strongly correlated with the IL12 secreted by the same gammadelta TCR+ cells, supporting the paradigm. Furthermore, IFNbeta1a therapy decreased the gammadelta TCR+ cell secretion of TH1 cytokines after 8-10 weeks of therapy. PMID- 11403228 TI - Peripheral blood T lymphocytes from systemic sclerosis patients show both Th1 and Th2 activation. AB - Our objective was to investigate the phenotype of helper T cells in the peripheral blood of patients with systemic sclerosis (SSc). PBMC from 15 patients with SSc and 15 sex- and age-matched controls were investigated for lymphocyte subpopulations (CD3, CD4, CD8, CD19, CD16/CD56, CD3-DR); IL-2, IL-4, and IFN gamma mRNAs; and the relative cytokines in their cytoplasm. The last assay was carried out both in unstimulated and in PMA-activated PBMC. SSc patients presented a higher percentage of activated T cells, CD3+ DR+ (19.7 +/- 9.9 vs 5.1 +/- 2.5%; P < 0.0001); 12 of them presented IFN-gamma mRNA-positive cells; and none IL-2 or IL-4 mRNAs. Under basal conditions, PBMC from six SSc patients contained IL-2, IL-4, and IFN-gamma (i.e., they showed both Th1 and Th2 activation), and 1 IFN-gamma only. PMA-stimulated PBMC of patients differed from those of controls only in the increased percentage of IFN-gamma positive cells (52 +/- 12 vs 37 +/- 11%; P < 0.01). Our study demonstrates that Thl activation occurs in the peripheral blood of SSc patients. This evidence must be faced with from both a pathogenetic and a therapeutical point of view. PMID- 11403229 TI - OKT3 and IL-2 treatment for purging of the latent HIV-1 reservoir in vivo results in selective long-lasting CD4+ T cell depletion. AB - Activation of resting T cells has been proposed to purge the reservoir of HIV-1 infected resting CD4+ T cells. We therefore treated three HIV-1-infected patients on antiretroviral therapy with OKT3, a CD3 monoclonal antibody, and recombinant human IL-2. Here we report the profound and partially long-lasting host responses induced by the OKT3 and IL-2 treatment. OKT3/IL-2 induced a strong but transient release of plasma cytokines and chemokines. The percentage CD4+ and CD8+ cells in the blood expressing the activation marker CD38 transiently increased to almost 100%, and in lymph nodes we "observed" a 10-fold increase in the number of dividing Ki67+ cells and increased numbers of apoptotic cells. Following OKT3/IL 2 treatment, a long-lasting depletion of CD4+ cells in the peripheral blood and lymph nodes occurred, suggesting the physical deletion of these cells. Increases in CD4+T cell numbers during the two year followup period were due mainly to increased memory cell numbers. CD8+ cells were also depleted in the blood, but less severely in lymph nodes, and returned to baseline levels within several weeks. PMID- 11403231 TI - Controlled release of lysozyme from succinylated gelatin microspheres. AB - Gelatin was anionized to increase the carboxylic acid groups through succinylation. Succinylation of gelatin was performed using varying amounts of succinic anhydride. This gave various percentages of substitution. Lysozyme, a cationic antibacterial enzyme, which has important applications in the reduction of prosthetic valve endocarditis, was chosen as a model protein drug. Microspheres were prepared using unmodified gelatin and succinylated gelatin (SG) and lysozyme was incorporated into them. The percentage loading and release profiles of lysozyme for gelatin and SG microspheres were evaluated and compared. It was found that the SG microspheres exhibited higher loading efficiency for lysozyme (50%) than the unmodified gelatin microspheres. The in vitro release of lysozyme from SG microspheres occurred up to 122 h, compared to 96 h for gelatin microspheres, for the release of most of the lysozyme incorporated. This prolonged release of lysozyme from SG microspheres was attributed to the electrostatic interaction between the cationic lysozyme and the anionic SG microsphere carrier. PMID- 11403230 TI - Evidence for a correlation between antibody-dependent cellular cytotoxicity mediating anti-HIV-1 antibodies and prognostic predictors of HIV infection. AB - Using our gp120/41-expressing, NK cell activity-resistant CEM.NKR cell clones as targets in HIV-1-specific antibody-dependent cellular cytotoxicity (ADCC) assays, we demonstrate here that the serum titers of anti-HIV-1 ADCC antibodies bear a significant (P < 0.05) positive correlation with the peripheral blood CD4+ T cell counts and a negative one with the number of copies of HIV-1 RNA in the plasma of HIV-infected individuals. These findings underscore the importance of these antibodies as a protective immune parameter in these infections. PMID- 11403232 TI - HSP 47 and collagen mRNA expression in L929 cells adhered to lipid films. AB - L929 cell adhesion on various lipid films prepared by Langmuir-Blodgett methods (LB method) were studied. L929 cells adhered to every lipid film similar to tissue culture poly(styrene) (TCPS). The mRNA expression of both collagen and HSP47 in adherent cells were evaluated by the RT-PCR method. mRNA expression of collagen was not altered during adhesion and proliferation duration, while HSP47 mRNA expression changed depending on culture time. L929 cells adhered to L-alpha dipalmitoylphosphatidylcholine (DPPC)-films showed little HSP47 mRNA expression. It was suggested that DPPC films regulate L929 cell function via unique serum protein adsorption. PMID- 11403233 TI - The compressive deformation of multicomponent microcapsules: influence of size, membrane thickness, and compression speed. AB - The clinical application of microcapsules for the immunoisolation of living tissue requires knowledge about the mechanical stability of polymer membranes. Microcapsules of 400-1000 microm in diameter were formed through the gelation of sodium alginate/sodium cellulose sulfate droplets through calcium chloride, with the membrane produced via complex coacervation between polyanions and poly(methylene-co-guanidine) hydrochloride. The deformation behavior of these multicomponent microcapsules was investigated by uniaxial compression experiments. Specifically, the influence of the deformation speed, capsule diameter, and membrane thickness on the mechanical properties was evaluated. The bursting force was found to be dependent on the deformation speed. Therefore, the measurement of the bursting work, a speed-independent value of the resistance to high stresses and deformations, was recommended as the most valid for capsule mechanical resistance. Furthermore, the bursting force was positively correlated with membrane thickness only for membrane-radius ratios up to 20%. For thicker membranes, the bursting event occurred because the opposite membranes touched each other, and not, because of insufficient strength. Indeed, the resistance to smaller deformations was positively correlated to the membrane thickness over the whole range of membrane-radius ratios. Moreover, the forces for constant deformation were linearly correlated to the total membrane volume, independently of capsule size and membrane thickness. PMID- 11403234 TI - Evaluation of bioresorbable polymers of lactic acid in a culture of human bone marrow cells. AB - Long term cultures of human bone marrow cells on poly(L-lactide-co-D,L-lactide) 70:30 and 90:10 plates were observed by means of scanning electron microscopy (SEM), SEM-EDX (SEM combined with energy dispersive X-ray analysis), flow cytometry, histochemical stainings, and culture medium analysis. After 14 days culture, cell numbers were only slightly lower compared with our reference material, hydroxyapatite, and much higher compared with polyethylene. There was evidence of collagenous matrix production with osteoblast activity. Acridine orange stainings as well as flow cytometry after incubation with propidium iodide showed only a few non-viable cells. By means of flow cytometry, we found about 30% of cells with granulocyte-markers, some monocyte-derived cells, and only small amounts of lymphocytes. After 9 weeks culture, there was evidence of calcium-phosphate deposition with extracellular matrix. There were only slight differences between the two tested polymers. Our culture system with human bone marrow cells plated on two bioresorbable polymers suggests a biocompatibility almost as good as hydroxyapatite, which is usually well tolerated. There was even evidence of mineralized collagenous matrix after some weeks of culture, which was detected earlier than the mineralization of cell-free controls. PMID- 11403235 TI - Degradation behavior of composite pins made of tricalcium phosphate and poly(L,DL lactide). AB - Combining a bioactive ceramic with a resorbable polymer may improve the biocompatibility and the osseointegration of degradable fracture fixation devices. This study reports on the mechanical properties and degradation behavior of two composite pins made of poly(L,DL-lactide) and 10 and 30% beta-tricalcium phosphate (TCP), respectively. The pins were compared to a pin made of 100% poly(L,DL-lactide). The failure force, bending strength, and molecular weight were determined during in vitro degradation at 37 degrees C up to 78 weeks. The blending with 10 or 30% beta-TCP decreased the initial mechanical properties and led to an accelerated degradation rate. The pins with 30% TCP lost half of their strength after 16 weeks, i.e. faster than the unmodified pin (40 weeks). The pins with 10% TCP, however, showed a decreased initial strength (128+/-9 MPa) compared to the unmodified pin (152+/-9 MPa) but very similar degradation characteristics. The drop of the molecular weight was not significantly different between the three types of pins. It was concluded that the mechanical requirements for a pin for the fixation of small bony fragments with improved biocompatibility were fulfilled by the composite pin with 10% TCP but not by the pin with 30% TCP. PMID- 11403236 TI - Biodegradable polyhydroxyalkanoate implants for osteomyelitis therapy: in vitro antibiotic release. AB - Various random copolyesters of 3-hydroxybutyrate and 3-hydroxyvalerate (PHBV) and 3-hydroxybutyrate and 4-hydroxybutyrate P(3HB-4HB) were used in the construction of biodegradable, implantable rods for the local delivery of antibiotics (Sulperazone and Duocid) in chronic osteomyelitis therapy. Drug loading, type of active agent, and additional coating of the implant surface all have significant contributions to the in vitro release profile. The rate and duration of Sulperazone release from P(3HB-4HB) rods were controlled by the polymer/drug ratio (drug loading). The rate of drug dissolution was substantially higher than that of polymer degradation. Therefore, the release phenomenon was more dependent on drug dissolution rather than on polymer degradation or diffusion. Coating rods with the same type of polymer substantially reduced the initial burst effect observed with the uncoated rods, and significantly decreased the release rate so that the release kinetics became almost zero order. Antibiotic release from coated rods was sustained for over a period of 2 weeks at a constant rate, whereas uncoated rods released their contents in less than a week. Impregnation of Duocid into the hydrophobic polymer matrix yielded a rod with a smoother surface topography. The release from these rods was significantly higher than for rods loaded with Sulperazone and a zero order release could not be obtained with these samples. PMID- 11403237 TI - Complex associates of plasmid DNA and a novel class of block copolymers with PEG and cationic segments as new vectors for gene delivery. AB - Cationic block copolymers, consisting of a poly(ethylene glycol) block and a block deriving from the poly(dimethylamino)ethyl methacrylate were prepared via a two-step procedure, based on the use of macroinitiators. By appropriately changing the experimental conditions and reacting the poly(dimethylamino)ethyl methacrylate block with iodo- or bromo-alkyl derivatives, a variety of ionic block copolymers with tuned physicochemical properties were prepared. These block copolymers are able to spontaneously self-assemble with plasmid DNA to produce oriented and shielded vectors, with physicochemical properties appropriate for in vivo applications. In addition, the formation of a complex between the cationic block copolymer and the plasmid DNA results in a nuclease resistance increase due to the stable nature of the complex. PMID- 11403238 TI - Phase transition and elasticity of protein-based hydrogels. AB - Reported are specific materials characterizations of three protein-based polymers comprised of repeating pentapeptide sequences, namely (GVGVP)251, (GVGIP)260 and (GVGVP GVGVP GEGVP GVGVP GVGVP GVGVP)n](GVGVP) where G = glycine, V = valine, P = proline, I = isoleucine, and E = glutamic acid, which had been previously prepared and gamma-irradiation cross-linked into elastic matrices. These polymers exhibit a hydrophobic folding and assembling transition on raising the temperature above a critical temperature, designated by Tt. Their equilibrium swelling ratio, uniaxial tensile and dynamic shear behavior were studied. The effect of the transition on swelling and mechanical properties was demonstrated. Equilibrium swelling ratio below the transition temperature decreased with the increase of gamma-irradiation dose. Above the transition temperature, hysteresis and frequency dependence were found in tensile loading-unloading tests and dynamic shear measurements, respectively. The rubber elasticity theory of random chain networks was applied to the data only below the transition temperature, where they may properly be considered random network hydrogels, to estimate molecular weight between cross-links and the Flory-Huggins interaction parameter. PMID- 11403239 TI - Interleukin-6 expression by osteoblast-like MG63 cells challenged with four acrylic bone cements. AB - Periprosthetic osteolysis is a major clinical problem in total hip and total knee arthroplasty and polymethylmethacrylate (PMMA) is a possible etiologic factor. Recently, increasing importance was ascribed to interleukin-6 (IL-6) as an agent favouring bone resorption. The aim of the present study was to investigate the role of bone cements on IL-6 production by MG63. The effect of four acrylic bone cements (Sulfix-60, CMW 1, CMW 2, and CMW 3) on the protein release and mRNA expression of IL-6 in osteoblast-like cell line MG63 was examined using IL-1beta (0.2 microg ml(-1)) as the positive control. The extracts in minimum essential medium (MEM) of the cements were tested, following 1-h and 7-day curing. CMW 1 and CMW 2 significantly increased the IL-6 release into the culture media (p < 0.01). The cells incubated with Sulfix-60 and CMW 3 produced no significantly different levels of IL-6 than the basal production. A positive correlation was found between the concentration of IL-6 and the contents of benzoylperoxide (p = 0.0003) and barium sulphate (p < 0.0001). MG63 expressed IL-6 mRNA constitutively, as demonstrated by the positivity of the negative controls too. We conclude that CMW 1 and CMW 2 increase the production of IL-6 in MG63 cells. The response to Sulfix-60 and CMW 3 is not significantly greater than the negative control. PMID- 11403240 TI - Partitioning of SDS in liposomes coated by the exopolymer excreted by Pseudoalteromonas antarctica NF3 as a measure of vesicle protection against this surfactant. AB - The capacity of glycoprotein (GP) excreted by Pseudoalteromonas antarctica NF3, to protect phosphatidylcholine (PC) liposomes against the action of the anionic surfactant sodium dodecyl sulfate (SDS) was studied in detail. To this end, changes in the surfactant partitioning between the lipid bilayer and the aqueous phase (partition coefficients, K) and in the effective surfactant to PC molar ratios (Re) were determined as a function of the amount of GP assembled with liposomes. The permeability of liposomes was determined by monitoring the changes in the fluorescence intensity of liposomes due to the release of the fluorescent dye 5(6)-carboxyfluorescein (CF) from the interior of vesicles to the bulk aqueous phase. Increasing GP amounts in the system resulted in the same interaction step as a rise in Re and a fall in the surfactant partitioning between the lipid bilayer and water. Hence, the higher the proportion of GP, the lower the surfactant ability to alter the permeability of liposomes and the lower its affinity with these bilayer structures. In addition, increasing GP proportions resulted in the same interaction step as a progressive increase of the free surfactant concentration (S(W)). The fact that the S(W) was always lower than the surfactant critical micelle concentration indicates that the interaction of SDS with coated liposomes was mainly ruled by the action of surfactant monomers in all cases. PMID- 11403241 TI - Differential neutrophil traffic in gut and lung after scald injury. AB - Neutrophil recruitment to the lung after thermal injury has been reported by various laboratories. Changes in neutrophil populations in the gut and lung have not been examined simultaneously after burn injury. Mice aged 8 to 10 weeks were anesthetized and subjected to 15% topical scald injury. Animals were sacrificed at 30 minutes and 1, 2, 4, 8, 16, and 32 hours after injury with harvesting of terminal ileum and lung for myeloperoxidase (MPO) assay. Lungs were evaluated after bronchoalveolar lavage and lavage of the vascular bed to remove neutrophils in these compartments. Myeloperoxidase activity was compared between groups of sham-injured and burned animals. Although pulmonary neutrophil recruitment was obvious after scald burn; in the ileum, burned animals showed diminished MPO activity. Histology and bronchoalveolar lavage revealed no evidence of gross organ injury apart from obvious changes in cellular content in the lung. Thermal injury is associated with differential neutrophil movement in the lung and the gut in this model. Pulmonary neutrophil recruitment is confirmed, whereas the gut seems to lose neutrophils as indicated by diminished MPO activity in the initial hours after dorsal scald injury. PMID- 11403242 TI - The effectiveness of risperidone on acute stress symptoms in adult burn patients: a preliminary retrospective pilot study. AB - The traumatic event of a burn injury can precipitate acute stress symptoms of nightmares, flashbacks, hyperarousal and disturbed sleep patterns. In some acutely burned hospitalized patients these symptoms may be significantly distressing, not respond to brief psychotherapeutic interventions, and may require pharmacotherapy. This regional burn center has seen clinically positive results with a small sample of patients using low doses of risperidone. In light of these clinical observations a preliminary retrospective pilot study was undertaken. Ten patients fit the criterion of having clinically significant distressing acute stress symptoms and were treated with risperidone. All 10 reported symptom diminishment or relief 1 to 2 days after starting risperidone. Improvement was defined as decreased sleep disturbances, diminished nightmares/flashbacks, and decreased hyperarousal. None of the patients reported any side effects from the low dose of risperidone (0.5-2 mg at bedtime; average dose was 1 mg). Patients with less clinically distressing symptoms were treated with supportive therapy and guided imagery. Results need to be interpreted cautiously because of the small sample size and lack of a control group. However, the results are encouraging enough to warrant a prospective study in order to better understand the efficacy of the use of risperidone in treating such symptoms in burn patients. PMID- 11403243 TI - The effects of exercise programming vs traditional outpatient therapy in the rehabilitation of severely burned children. AB - The purpose of this study was to compare the efficacy and effects of exercise programming (Study group, n = 11) vs traditional outpatient therapy (Home group, n = 10) in burned children (> 40% body surface area). This was a prospective, randomized, controlled trial in a hospital-based children's wellness center. Twenty-one patients (13 boys and 8 girls) averaging 10.6 +/- 0.9 years and TBSA = 59.7 +/- 3.1% were evaluated 6 and 9 months postburn. Moderate intensity, progressive resistance and aerobic exercise conducted 3 times weekly for 1 hour were a supplement to standard therapy over 12 weeks. Muscular strength and functional outcome significantly increased in both groups (P < .05). Improvements in strength (80.1 vs 37.7%) and distance walked (39.5 vs 12.5%) were significantly greater for Study vs Home groups, respectively, P < .05. We conclude that exercise programming may be safely included in rehabilitation programs for severely burned children and can be effective in increasing muscular strength and functional outcome. PMID- 11403244 TI - Community integration after burn injuries. AB - Evaluation of community integration is a meaningful outcome criterion after major burn injury. The Community Integration Questionnaire (CIQ) was administered to 463 individuals with major burn injuries. The CIQ results in Total, Home Integration, Social Integration, and Productivity scores. The purposes of this study were to determine change in CIQ scores over time and what burn injury and demographic factors predict CIQ scores. The CIQ scores did not change significantly from 6 to 12 to 24 months postburn injury. Home integration scores were best predicted by sex and living situation; Social Integration scores by marital status; and Productivity scores by functional outcome, burn severity, age, and preburn work factors. The data demonstrate that individuals with burn injuries have significant difficulties with community integration due to burn and nonburn related factors. CIQ scores did not improve over time but improvement may have occurred before the initial 6-month postburn injury follow-up in this study. PMID- 11403245 TI - An analysis of 550 hospitalized pediatric burn patients in Hong Kong. AB - A total of 550 acute burn patients under the age of 15 years were admitted to the Burns Unit of Prince of Wales Hospital, Hong Kong between March 1993 and February 1999. There were 337 males and 213 females with a male to female ratio of 1.58:1. The median age was 2.5 years and the median burn size was 5% total body surface area (TBSA). Toddlers of age < 2 years accounted for 235 (42.7%) of admission. Domestic burns resulted in 481 (87.5%) injuries followed by play-related burns that caused 34 (6.2%) admissions. Scalding was the most common cause of injury, which accounted for 497 (90.4%) admissions, followed by flame burns, which resulted in 45 (8.2%) injuries. Nine patients (1.6%) had inhalation injury requiring intubation and ventilatory support. The median hospital stay was 8 days and there was no seasonal variation in admission. The majority of patients (80.2%) had their wounds healed without any operation. Only 19 out of 550 patients (3.5%) had burns of 30% TBSA or larger, and only nine patients (1.9%) had inhalation injuries. Only one patient died in this series, which yielded a mortality rate of 0.2%. PMID- 11403246 TI - Initial experience with a prototype staple detector. AB - Stainless steel staples can be overgrown by granulation tissue or skin grafts and become buried in tissue. Although they are generally asymptomatic, they can on rare occasion erode to the surface or complicate the radiographic evaluation of pain not related to the staples. A device to facilitate detection of retained staples would be desirable, but it has been difficult to develop because stainless steel staples have poor magnetic and conductive properties. A prototype device, based on the presence of metal disturbing a low-power electromagnetic field, was developed. With human studies committee approval, this miniaturized detector was used in 13 burn patients to detect staples during planned removal of large numbers of staples. Staples were first removed using visual inspection and palpation of the wounds. This was followed by use of the staple detector. The age of the patients was 10.8 +/- 3.3 years, and burn size was 54.6 +/- 8.8% of the body surface. In 8 (62%) of the patients one or more additional staples were detected by the device that would otherwise have been overlooked. In 4 (31%) of the patients there was a false-positive signal, possibly related to topical silver in the wounds, that required additional focused physical examination. A portable staple detector has been developed. Initial experience with the device is favorable and warrants an expanded trial, which is in the planning stages. PMID- 11403247 TI - Safety and pharmacokinetics of EMLA in the treatment of postburn pruritus in pediatric patients: a pilot study. AB - The purpose of this study was to determine the safety and pharmacokinetics of a eutectic mixture of local anesthetics (EMLA) used to ameliorate postburn pruritus after application onto newly formed, intact skin in children. EMLA was applied once to an itchy site where healed skin had formed. Serial blood samples were collected to measure lidocaine, prilocaine, o-toluidine, and methemoglobin. Maximal plasma concentration, minimal plasma concentration, time to achieve the maximal plasma concentration, elimination half-life, and area under the concentration-time curve were calculated. Vital signs, oxygen saturation, clinical signs of hypoxia, and itch intensity were measured. Five children had 15.7 +/- 2.54 g (+/- SD) of EMLA applied to a skin surface area of 93.0 +/- 37.0 cm2. Lidocaine and prilocaine concentrations were below toxic levels; o-toluidine was not detected. Methemoglobin remained between 1 and 3%; patients did not exhibit any clinical signs of hypoxia. Mean oxygen saturation was 98.9 +/- 0.01%. The mean number of pruritic episodes and antihistamine breakthrough doses were greater in the 2 prestudy control days compared with study day 3 (P = 0.01 and P = 0.03, respectively). Skin at the site of EMLA application remained anesthetized for 12 to 13 hours. In this small pilot study, EMLA seems to be a safe, novel treatment for postburn pruritus in burned children when applied to newly healed, intact skin. PMID- 11403248 TI - Burns and injuries resulting from the use of gel candles. AB - Scented gel candles are common decorative household items composed of gelled mineral oil, fragrances, and dye. Like traditional wax candles, they have an open flame. Because of defective design, there have been several burns and injuries caused by these products. Here we report our experience with a scald burn from a gel candle and describe 34 additional injuries attributed to gel candles previously unreported in the medical literature. PMID- 11403249 TI - Sodium azide burn: a case report. AB - Chemical burn injuries commonly occur at the workplace and can be caused by a variety of agents. Sodium azide is a volatile compound used in the industrial setting and it is also a constituent of car airbags. The known toxic effects of sodium azide include hypotension, bradycardia, and headaches. At the cellular level, it inhibits of ATP production by blocking the respiratory oxidation cascade. In the burn literature only one previous report documents a sodium azide hand burn caused by airbag malfunction. We report a case of massive exposure and resultant systemic toxicity from a sodium azide canister explosion. PMID- 11403250 TI - Management of an inpatient-outpatient clinic an eight-year review. AB - Healthcare organizations have historically separated outpatient from inpatient facilities. In order to streamline the continuity of high quality care, an outpatient burn clinic was established on our inpatient burn center in 1991. Management of the outpatient clinic required alternate staffing patterns and supply allocation plus training in managed care and third party payors. Budget decisions and health care trends affected the number of full time equivalents (FTEs). Between 1990 and 1998, a 33% RN FTE reduction occurred with an overall 22% decrease in total inpatient care providers. Clinic positions were allocated as patient volume and workload data could justify additional staff. Enhanced flexibility in assignment and use of personnel with varying skill levels led to efficient integration of inpatient and outpatient care with an overall reduction in RN FTEs. The purpose of this study is to review the changes in nursing management strategies required by this consolidation. PMID- 11403251 TI - A developmental perspective on psychological principles of burn care. AB - The treatment of pediatric burn patients requires careful attention to psychological principles underlying good pain management. These principles, and a variety of specific psychological treatments, have been described thoroughly. In this article the cognitive capabilities, emotional understanding, and behavioral competencies that characterize children at various stages of development are outlined. Important age-related developmental issues are noted, and the child's experience of a burn injury at different ages is outlined. Guidelines for interventions tailored to the developmental capacities and frailties of children in different age groups are offered. Various psychological coping techniques are understood in light of their likely value for pain management with a developmental framework in mind. PMID- 11403252 TI - The use of high frequency ultrasonography in the prediction of burn depth. PMID- 11403253 TI - CD8 T cell-mediated rejection of intestinal allografts is resistant to inhibition of the CD40/CD154 costimulatory pathway. AB - BACKGROUND: Disruption of the CD40/CD154 pathway inhibits rejection in numerous models. The importance of this pathway on intestinal allograft rejection was examined in this study. METHODS: Intestinal grafts from B6C3F1 mice transplanted into C57BL/6 recipients were assessed histologically for rejection. RESULTS: The monoclonal antibody to CD154, MR1, failed to inhibit rejection in wild-type mice. Similarly, CD154-/- recipient mice rejected intestinal allografts. MR1 did inhibit early rejection in CD8-/- mice, but had no effect in CD4-/- recipients. All MR1-treated CD8-/- recipients eventually developed rejection. No benefit was observed when blockade of the CD40/CD154 pathway by MR1 was combined with blockade of the CD28/B7 pathway by mCTLA4Ig. CONCLUSIONS: These data suggest that CD4+ T cells mediating intestinal allograft rejection may be more dependent upon the CD40/CD154 pathway than CD8+ T cells. This finding highlights the importance of identifying agents that suppress CD8+ T cell-mediated rejection. PMID- 11403254 TI - Cytodiagnosis of a meningeal fibrosarcoma metastatic to the thyroid gland. AB - A case of primary leptomeningeal fibrosarcoma metastatic to the thyroid gland, diagnosed by fine needle aspiration biopsy is described. The patient, a 39-year old women was initially diagnosed with a leptomeningeal fibrosarcoma. Sections of the primary tumor studied by electron microscopy showed that the tumor cells have the ultrastructure features of a fibroblast. She underwent tumor resection, followed by chemotherapy and local radiotherapy. Four months later, she presented with dysphagia. Ultrasonography confirmed the presence of a 2-cm mass in the right lobe of thyroid. A fine needle aspiration biopsy was performed. Light microscopy showed interwoven bundles of neoplastic spindle cells similar to the primary tumor, consistent with a metastatic fibrosarcoma. To our knowledge, this is the first report of a primary meningeal fibrosarcoma that metastasized to the thyroid gland, which was diagnosed by a fine needle aspiration biopsy. PMID- 11403255 TI - The use of fine needle aspiration biopsy in the evaluation of lymphadenopathy. AB - Fine needle aspiration biopsy is an important diagnostic tool in the evaluation and triage of patients with lymphadenopathy. It offers a simple and inexpensive test for diagnosis of reactive hyperplasia, infections, granulomatous lymphadenopathies, and metastatic diseases. Although previously regarded as limited in its use for diagnosing primary lymphoid malignancies, fine needle aspiration in combination with immunophenotypic and genotype studies is gaining respect in providing accurate diagnosis of lymphoma for primary treatment in selected patients. PMID- 11403256 TI - Diagnostic challenges in aspiration cytology of the salivary glands. AB - The main goal of fine-needle aspiration (FNA) of salivary gland lesions is to assist the clinician in the management of patients who present with a mass lesion. Cytologic examination aims to determine, if a process is inflammatory and/or reactive, benign or malignant neoplasm and if possible renders a specific diagnosis. It has been argued that in the area of salivary gland tumors, surgical management relies less heavily on a specific preoperative diagnosis, because almost all neoplastic salivary gland lesions will undergo surgical excision. However, knowing beforehand if a lesion is malignant or benign, will aid in planning surgery and may prompt or postpone decisions for surgical intervention. The salivary glands are unique in their histologic complexity and morphological variability of tumors, which is reflected in the cytologic material. In addition to the overlapping morphologic patterns of salivary gland tumors, they also represent relatively rare lesions, thus making it more difficult to acquire diagnostic expertise in FNA. Other than approaching salivary gland tumors by a description of single entities in their benign and malignant categories, we favor a more practical approach to diagnosis based on the key morphologic features noted in FNAs. This article addresses differential diagnoses according to the predominant cytologic presentation with attention to the cell type and size, nature of the cytoplasm, and the smear background. PMID- 11403257 TI - Parotid gland mucosa associated lymphomas and their cytologic mimics. AB - Although the parotid is a target for myoepithelial sialadenitis and lymphomas of mucosa associated lymphoid tissue (MALTomas), MALTomas are rare and have not been well characterized in the cytology literature. We examine fine needle aspiration biopsies from 19 lymphoid lesions of the parotid gland with particular emphasis on MALTomas and their cytologic look-a-likes. Purely on cytology it is difficult to distinguish with certainty a MALToma from a late-phase reactive lymph node. Both conditions were predominantly composed of small mature appearing lymphocytes and both occasionally had larger lymphocytes with tingible body macrophages. By contrast, the diagnosis of non-MALT lymphoma was usually straight forward, as they had overt cytologic atypia. In applying fine needle aspiration biopsy to large or persistent lymphoid lesions of extra-nodal sites like the parotid gland, we recommend the liberal use of flow cytometric analysis to distinguish between reactive lymph nodes and MALToma and to more precisely classify non-MALT lymphomas. PMID- 11403258 TI - Thyroid aspiration cytology: a "cell pattern" approach to interpretation. AB - The key to the interpretation of thyroid fine needle aspiration is largely dependent on the recognition of various morphologic patterns of epithelial cells, usually follicular cells, and background elements, such as colloid. These morphologic patterns consist of 3 parts: 1) The arrangement of cells with respect to one another, 2) The cytologic features of individual cells, and 3) The presence of background elements. The cellular arrangements generally encountered in fine needle aspiration of the thyroid include the follicular patterns (macro /normo-follicular and micro-follicular), the papillary pattern, the syncytial pattern, the dispersed cell pattern, and the cystic pattern. This article approaches some of the differential diagnostic challenges encountered while interpreting thyroid aspiration cytology by focusing first on the overall cellular arrangement to generate a differential diagnosis and then narrowing that differential by assessing cellular features of individual cells and the presence of background elements. PMID- 11403259 TI - Concomitant papillary thyroid carcinoma and Hashimoto's thyroiditis. AB - An association between papillary thyroid carcinoma (PTC) and Hashimoto's thyroiditis (HT) is well recognized. Both entities may often display overlapping morphologic features. The aim of this study was to evaluate the accuracy of fine needle aspiration (FNA) of concomitant PTC and HT. Twenty nine thyroid FNAs with a diagnosis of concomitant PTC and HT on follow-up surgical material were retrospectively reviewed (11% of all HT cases diagnosed in the same period of time). The cytologic specimens were evaluated for the presence of diagnostic features of PTC and HT. In 16 of 29 cases, the diagnosis of PTC was made or suggested; however, only in 3 cases were both entities recognized on the FNA material. The review of the remaining cases (13 cases) showed diagnostic features of PTC in 2 cases (interpretation errors), some features of PTC in 8 cases (insufficient diagnostic features), features of only HT in 2 cases, and 1 case was acellular (sampling errors). Originally, 10 cases with features of PTC were diagnosed as either follicular neoplasm or colloid nodule with or without HT. Histologically, 1 of 13 cases was a cystic variant and 7 of 13 cases were follicular variants of papillary carcinoma. It is important to be aware of the coexistence of PTC and HT. Deliberate search for evidences of PTC in every case of HT may be necessary to improve diagnostic accuracy of the FNA. However, the cytologic diagnosis of follicular variant of PTC coexisting with HT can be challenging. The sampling error may also cause false negative results. PMID- 11403260 TI - Antinuclear antibodies: to penetrate or not to penetrate, that was the question. PMID- 11403261 TI - Peripheral blood CD4+ T cells in systemic lupus erythematosus. PMID- 11403262 TI - CD4 positive peripheral T cells from patients with systemic lupus erythematosus (SLE) are clonally expanded. AB - T cell activation was analysed in peripheral CD4+ T cells from both systemic lupus erythematosus (SLE) patients with active and inactive disease as well as in normal healthy donors (NHD) to investigate the involvement of CD4+ T cells in the etiopathogenesis of SLE. CD4+ T cell receptor (TCR) beta-chain transcripts, containing the complementarity determining region 3 (CDR3), were amplified by reverse transcriptase-polymerase chain reaction (RT-PCR) and analysed by high resolution polyacrylamide gel electrophoresis. In addition the CDR3 of both clonally activated as well as heterogeneous Vbeta families from SLE patients were analysed at the molecular level. We observed a restricted CDR3 length polymorphism in peripheral CD4+ T cells from SLE patients compared with NHD, more pronounced in patients with high disease activity. Furthermore, in some Vbeta families single peaks in the histogram indicated nearly monoclonal T cell expansion. Sequencing of selected TCR beta-chains revealed a increased content of acidic amino acids in the CDR3 encoded by either proximal Jbeta elements or N nucleotides. We conclude that CD4+ T cells from peripheral blood of SLE patients display features of a secondary antigen driven immune response. The bias of the CDR3 towards acidic amino acids suggests the involvement of positively charged antigens. PMID- 11403263 TI - Counter-proliferative effects of nucleosomal antigens in cultures from lupus patients. AB - Blood mononuclear cells from 20 lupus patients were cultured in the presence of nucleosomal antigens to determine whether they induce lymphocyte proliferation. The predominant effect seen, however, was one of inhibition of the background proliferation. Such inhibition was rare with cells from female or male controls. Nucleohistone (NH), crude histone and enriched preparations of histones H2A/H4, H2B and H3 showed this effect in approximately one-third of patients, but H1 and single-stranded (ss) DNA had no such activity. Double-stranded (ds) DNA may show this inhibitory action, but further tests are required. ssDNA was the only antigen that showed evidence (two patients) of disease-related stimulation of proliferation. Histones and NH induced proliferation in many subjects but the strongest responders were controls. Patients responded poorly to tuberculin PPD but gave an exceptionally strong proliferative response to pokeweed mitogen. It is suggested that the inhibition of background proliferation in patients is a consequence of the interaction of nucleosomal antigens with sensitised T cells. If T cell sensitisation to histones is an important factor in the development of lupus, the disease may be preventable in those at risk by inducing tolerance to the appropriate peptides. PMID- 11403264 TI - Detection of macroprolactinemia with the polyethylene glycol precipitation test in systemic lupus erythematosus patients with hyperprolactinemia. AB - The objective of this study was to determine the diagnostic performance of the percentage of serum prolactin (PRL) precipitated with polyethylene glycol (PEG) for the detection of macroprolactinemia in systemic lupus erythematosus (SLE) patients with hyperprolactinemia. Serum samples from SLE patients were examined. Serum PRL was measured by immunoradiometric assay (IRMA) and samples with hyperprolactinemia (> 20 ng/ml) were submitted to PEG precipitation, gel filtration chromatography and affinity chromatography with protein-G sepharose. A comparative survey was used. Among 259 consecutive serum samples from SLE patients, PRL was > 20.1 ng/ml in 43 samples (16.6%). Gel filtration showed a predominant pattern of macroprolactinemia (> 100 kDa) in 14 (32.6%), a predominant pattern of monomeric PRL (23 kDa) in 27 (62.7%), and a variable pattern in two (4.7%). All sera with a predominant pattern of macroprolactinemia displayed anti-PRL autoantibodies by affinity chromatography for IgG. The best cut-off point for percentage of serum PRL precipitated with PEG for detection of macroprolactinemia was > or = 58.4%. Sensitivity and specificity were 100 and 96.6%, respectively. We can conclude that PEG precipitation is a convenient and simple procedure to screen for the presence of macroprolactinemia in sera from SLE patients. Precipitations > or = 58.4% are indicative of the presence of, and those < 50% the absence of, macroprolactinemia. However, samples with precipitations between 50 and 58.3% require gel filtration chromatography to characterize the predominant molecular form of PRL. Therefore, it is important to take these findings into account in future studies that aim to establish a relationship between PRL and disease activity in SLE. PMID- 11403265 TI - Anti-oxidized low-density-lipoprotein (OxLDL) antibodies in systemic lupus erythematosus with and without antiphospholipid syndrome. AB - The aim of this study was to examine potential links between antiOxLDL antibodies and the clinical and biological features of secondary antiphospholipid syndrome (II APLS) associated with systemic lupus erythematosus (SLE). A cohort study was done of 98 SLE patients followed-up for 1 y, including 18 with definite II APLS and 13 patients with definite primary APLS (I APLS). IgG anticardiolipin, IgG anti beta2 GPI, lupus anticoagulant, VDRL and IgG antiOxLDL were measured in all 98 study subjects. High antiOxLDL titers were found in seven (39%) of the 18 patients with II APLS vs 10 (12.5%) of the 80 patients without APLS (P < 0.01; OR = 4.45; 95% CI = 1.4-14.1) and none of the 13 patients with I APLS (P < 0.02). The mean antiOxLDL titer was not significantly higher in the SLE patients with than without II APLS (P > 0.05). A high antiOxLDL titer was correlated with deep venous thrombosis (P < 0.01; OR = 5.77; 95% CI = 0.54-61) but not with arterial thrombosis (P > 0.05; OR = 1; 95% CI = 0.29-3.09), thrombocytopenia, central nervous system involvement, livedo reticularis, or a positive Coombs test. The antiOxLDL antibody titer was correlated with the IgG anticardiolipin antibody titer (r = 0.235; P = 0.02) and with the IgG anti-beta2 GPI antibody titer (r = 0.224; P = 0.026). AntiOxLDL elevation was found in 17% of SLE patients and was significantly associated with II APLS and venous thrombosis. We found no evidence suggesting that antiOxLDL may be associated with atherosclerosis. PMID- 11403266 TI - Disassociation of sex hormone levels and cytokine production in SLE patients. AB - This study examines whether changes in the cytokine milieu of patients with systemic lupus erythematosus (SLE) are associated with abnormal levels of sex hormone levels in serum. The concentration of 17beta-estradiol (E2), progesterone (Pg) and dehydroepiandrosterone-sulphate (DHEAS) was monitored in sera from 128 lupus patients and 96 controls, and correlated with the activity of their cytokine secreting cells. Results indicate that SLE patients have (i) significantly fewer cells secreting IFNgamma, (ii) increased serum E2 and Pg levels, and (iii) reduced serum DHEAS levels compared to normal controls. However, the observed abnormalities in the cytokine milieu of SLE patients did not correlate with abnormalities in serum sex hormone levels. Instead, the association between IFNgamma production and DHEAS levels evident in healthy controls is absent in SLE patients, suggesting that cells from lupus patients are defective in their ability to produce IFNgamma in response to physiologic stimuli. Similarly, the normal correlation between IL-4 production and E2 levels was lost in patients with severe disease. Thus, while it remains possible that increased E2 and reduced DHEAS levels in lupus patients may help induce cytokine abnormalities early in disease, the subsequent cytokine imbalance does not correlate with sex hormone levels. PMID- 11403267 TI - Lipid and lipoprotein levels in premenopausal systemic lupus erythematosus patients. AB - The purpose of this study was to assess the prevalence of dyslipoproteinemia and to analyze the clinical variables that are associated with it in a sample of premenopausal systemic lupus erythematosus (SLE) patients. We studied 53 premenopausal (34.5 y) SLE outpatients and 45 controls. Clinical variables studied included patient age, weight, height, body mass index (BMI), age at disease onset, disease duration, clinical activity of SLE, renal involvement and drug therapy. Total cholesterol (TC), high- and low-density lipoprotein cholesterol (HDL-C and LDL-C), and triglycerides were measured using standard enzymatic techniques. Apolipoproteins (apo) A-I and B were determined by radial immunodiffusion. Twenty-nine patients (55%) and 14 controls (30%) had dyslipoproteinemia. An increase in TC, triglycerides, HDL3-C, apo A-I and apo B, and a decrease in HDL2-C and HDL-C/TC index was found in SLE patients in comparison with controls. TC (P = 0.007), apo B (P = 0.02), LDL-C (P = 0.03) and triglycerides (P = 0.0001) were significantly correlated with proteinuria. Patients on prednisone therapy had higher triglycerides levels (P = 0.03) than untreated patients. TC (P = 0.01), LDL-C (P = 0.006) and triglycerides (P = 0.04) were also correlated with the dose of prednisone. Dyslipoproteinemia is a common feature in adult SLE premenopausal patients which is characterized by an increase in TC, triglycerides and apo B, and an abnormal distribution of HDL subclasses. Corticosteroid therapy and proteinuria are the best predictors of dyslipoproteinemia in these patients. PMID- 11403268 TI - Skin lesions--an indicator of disease activity in systemic lupus erythematosus? AB - Cutaneous manifestations have great diagnostic value for systemic lupus erythematosus (SLE). In this study we tried to establish a correlation between lupus erythematosus LE-specific and LE-nonspecific cutaneous lesions and disease activity measured by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Sixty-six patients with SLE were evaluated. They were divided into three groups having: (1) only LE-specific lesions (38 or 58.46%); (2) only LE nonspecific lesions (4 or 6.15%); and (3) both types of lesions (23 or 35.38%). Results were analyzed using the Student t-test. Patients with LE-nonspecific skin manifestations had significantly increased disease activity compared to those with only LE-specific lesions. The number of different skin lesion types also correlated with disease activity. It was significantly increased in a group with three different types of lesion, either specific or nonspecific. Patients with only one type of lesion had mild disease. An intermediate disease activity was found in the group with two different lesion types. Lupus-specific skin manifestations serve primarily as an important diagnostic clue. In conclusion, patients with LE-nonspecific lesions have significantly more active SLE than those with LE-specific lesions and may therefore require more intensive therapy and disease monitoring. PMID- 11403269 TI - Pinguecula and Sjogren's syndrome: two cases. AB - A pinguecula is a small, raised conjunctival nodule at the temporal or nasal limbus classically associated with conjunctival microtrauma from exposure to sunlight and/or to dust. We report two cases occurring during the course of Sjogren's syndrome. To our knowledge this association has never been described. PMID- 11403270 TI - Antiphospholipid syndrome induced by HIV. AB - A 24-y-old male who developed necrotic lesions on the lower extremities together with testicular thrombosis necessitating orchiectomy, demonstrated high level IgG anticardiolipin (aCL) associated with acute HIV infection. This is one of the first cases describing a close relationship between viral infection and the classic antiphospholipid syndrome (APS). It is well documented that HIV patients may produce antiphospholipid antibodies (aPL), but the full-blown picture of the APS is distinctly uncommon with HIV or any other viral infection, possibly due to the overproduction of the IgM isotype rather than IgG aCL as in this case. The induction of thrombosis following infections has been well described in patients with catastrophic antiphospholipid syndrome (CAPS) but not in patients with the 'classic or simple' APS. PMID- 11403271 TI - The Systemic Lupus International Collaborating Clinics Group--origins and outcomes. PMID- 11403272 TI - Lupus nephritis in Lebanon. AB - This is a retrospective study of the clinicopathological characteristics of 50 systemic lupus erythematosus patients with nephritis who underwent a kidney biopsy and were admitted to the American University of Beirut Medical Center, in Lebanon, between 1979 and 1999. There were 43 females and seven males, with a median age of 24 y. Renal histology slides from these patients were assessed according to the World Health Organization classification, and were distributed as follows: class I (n = 3, 6%); class II (n = 14, 28%); class III (n = 11, 22%); class IV (n = 19, 38%); class V (n = 1, 2%); class VI (n = 2, 4%). All the patients received oral prednisone, in addition the following treatments were used: pulse intravenous (i.v.) cyclophosphamide (n = 23, 46%); azathioprine (n = 22, 44%); pulse i.v. steroids (n = 19, 38%); chloroquine sulfate (n = 17, 34%); methotrexate (n = 5, 10%); and plasmapheresis (n = 2, 4%). The median duration of follow-up was 5 y (range 1-33 y). On their last evaluation, out of 37 patients who were followed, 20 patients (54%) had controlled disease, eight patients (22%) were still on active medical treatment, four patients (11%) were on chronic hemodialysis, and five patients (13%) had died. Unlike three other Arab populations studies from Kuwait, United Arab Emirates and Saudi Arabia, where the most frequent histopathologic abnormality was class III, diffuse proliferative LN (class IV) was the most common type of lupus nephritis in Lebanon, similarly to reports from USA, France, Netherlands, South Africa, Thailand and Taiwan. PMID- 11403273 TI - ARDS in catastrophic antiphospholipid syndrome. PMID- 11403274 TI - Analysis of cocaine, benzoylecgonine, codeine, and morphine in hair by supercritical fluid extraction with carbon dioxide modified with methanol. AB - Supercritical fluid extraction (SFE) using CO2 modified with methanol (10%) was employed for the extraction of cocaine from human hair. Extraction conditions were developed by using designed experiments to characterize the effects of pressure, temperature, and percent methanol on the recovery of cocaine from hair. When compared to the conventional acid hydrolysis method for hair analysis, SFE was faster and gave higher recoveries. Amounts of cocaine, benzoylecgonine (cocaine metabolite), codeine, and morphine in a hair standard reference material were determined by SFE-gas chromatography/ mass spectrometry (GC/MS) and found to be in agreement with reported levels. Analyses of hair from forensic case studies are also reported. PMID- 11403275 TI - Ion-pair RP-HPLC determination of sugars, amino sugars, and uronic acids after derivatization with p-aminobenzoic acid. AB - A new, selective, and sensitive ion-pair RP-HPLC method for the simultaneous determination of three classes of natural organic compounds, i.e., carbohydrates, amino sugars, and uronic acids, in environmental samples is presented. p Aminobenzoic acid is used for precolumn derivatization of the analytes, enabling fluorescence (lambda(ex) 313 nm, lambda(em) 358 nm) or photometric detection (303 nm). The dependence of the derivatization yield on the reaction conditions is examined. Derivatives of lactose, galactose, glucose, mannose, xylose, arabinose, galacturonic acid, glucuronic acid, N-acetylglucosamine, and glycerinealdehyde were separated on a RP-C18 column with hydrophilic end capping within 35 min, applying TBAHSO4 as the ion-pair reagent. The concentration detection limits range between 20 and 30 microg L(-1) ((1-2) x 10(-7) M) for fluorescence detection and between 30 and 75 microg L(-1) for UV detection. A good linearity is achieved in the concentration range from 50 microg L(-1) to 100 mg L(-1) (r2 > 0.99). The described method has been applied for the determination of mono /disaccharides, uronic acids, and amino sugars in soil solutions and in landfill leachates. PMID- 11403276 TI - In-tube moleculary imprinted polymer solid-phase microextraction for the selective determination of propranolol. AB - A molecularly imprinted polymer (MIP) material was synthesized for use as an in tube solid-phase microextraction (SPME) adsorbent. The inherent selectivity and chemical and physical robustness of the MIP material was demonstrated as an effective stationary-phase material for in-tube SPME. An automated and on-line MIP SPME extraction method was developed for propranolol determination in biological fluids. This simplified the sample preparation process and the chromatographic separation of several beta-blocker compounds. The method developed for propranolol showed improved selectivity in comparison to alternative in-tube stationary-phase materials, overcoming the limitations of existing SPME coating materials. Preconcentration of the sample by the MIP adsorbent increased the sensitivity, yielding a limit of detection of 0.32 microg/mL by UV detection. Excellent method reproducibility (RSD < 5.0%) and column reusability (> 500 injections) were observed over a fairly wide linear dynamic range (0.5-100 microg/mL) in serum samples. To our knowledge, this is the first report on the automated application of a MIP material for in-tube SPME. The method was inexpensive, simple to set up, and simplified the choice of SPME adsorbent for in-tube extraction. The approach can potentially be extended to other MIPs for the determination of a wide range of chemically significant analytes. PMID- 11403277 TI - High-performance liquid chromatography-electrospray ionization mass spectrometry using monolithic capillary columns for proteomic studies. AB - The use of tetrahydrofuran/decanol as porogens for the fabrication of micropellicular poly(styrene/divinylbenzene) monoliths enabled the rapid and highly efficient separation of peptides and proteins by reversed-phase high performance liquid chromatography (RP-HPLC). In contrast to conventional, granular, porous stationary phases, in which the loading capacity is a function of molecular mass, the loadability of the monoliths both for small peptides and large proteins was within the 0.40.9-pmol range for a 60- x 0.2-mm capillary column. Lower limits of detection obtained by measuring UV-absorbance at 214 nm with a 3-nl capillary detection cell were 500 amol for an octapeptide and 200 amol for ribonuclease A. Upon reduction of the concentration of trifluoroacetic acid in the eluent from the commonly used 0.1-0.2 to 0.05%, the separation system was successfully coupled to electrospray ionization mass spectrometry (ESI-MS) at the cost of only a small decrease in separation efficiency. Detection limits for proteins with ESI-MS were in the lower femtomole range. High-quality mass spectra were extracted from the reconstructed ion chromatograms, from which the masses of both peptides and proteins were deduced at a mass accuracy of 50-150 ppm. The applicability of monolithic column technology in proteomics was demonstrated by the mass fingerprinting of tryptic peptides of bovine catalase and human transferrin and by the analysis of membrane proteins related to the photosystem II antenna complex of higher plants. PMID- 11403278 TI - Detection of in vivo formed DNA adducts at the part-per-billion level by capillary liquid chromatography/microelectrospray mass spectrometry. AB - Capillary liquid chromatography/microelectrospray mass spectrometry has been applied to the detection of deoxyribonucleoside adducts of the food-derived mutagen 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) from in vivo sources. Adjustments were made to a previously described methodology such that analyte detection could be improved by nearly 2 orders of magnitude. These adjustments included changing the electrospray ionization sprayer configuration, increasing the sample injection volume, improving the solid-phase extraction procedure, and increasing peak efficiency by modifying chromatographic conditions. While this scheme for improving analyte detection was targeted for DNA adducts, it could be applied to almost any LC/MS methodology where sensitive analysis is the primary objective. Selective reaction monitoring) techniques with a triple quadrupole mass spectrometer enabled sensitive and specific detection of IQ adducts, with detection limits approaching 1 adduct in 10(9) unmodified bases using approximately 500 microg of DNA. The DNA adducts N-(2'-deoxyguanosin-8-yl)-2 amino-3-methylimidazo[4,5-f]quinoline and 5-(2'-deoxyguanosin-N2-yl)-2-amino-3 methylimidazo[4,5-f]quinoline were detected in pancreas tissue of a cynomolgus monkey sacrificed 24 h after a single administration of 10 mg/kg carcinogen. The LC/MS results were consistent with previously published 32P-postlabeling data (Turesky et al. Chem Res. Toxicol. 1996, 9, 403-408). Thus, capillary tandem LC/MS is a highly sensitive technique, which can be used to screen for DNA adducts in vivo. PMID- 11403279 TI - Determination of vapor pressure of low-volatility compounds using a method to obtain saturated vapor with coated capillary columns. AB - The vapor pressures of O-ethyl S-2-diisopropylaminoethyl methylphosphonothiolate (VX), O-isobutyl S-2-diethylaminoethyl methylphosphonothiolate (RVX), and 2,4 dinitrotoluene (2,4-DNT) were determined with the gas saturation method in temperatures ranging from -12 to 103 degrees C. The saturated vapor was generated using a fused-silica column coated with the compound. This column was placed in a gas chromatograph, and the vapor pressure was determined directly from the detector signal or by sampling on Tenax tubes that were subsequently analyzed. From the linear relationships obtained by plotting log P vs 1/T, the enthalpies of vaporization (deltaHvap) and the vapor pressures at selected temperatures were determined. The vapor pressure of VX at 25 degrees C was 0.110 Pa and the deltaHvap 77.9 kJ x mol(-1). The corresponding results for RVX were 0.082 Pa and 76.6 kJ x mol(-1). The vapor pressure of 2,4-DNT at 72 degrees C (melting point) was determined to 6.0 Pa, and the enthalpies of the solid and the liquid state were 94.2 and 75.3 kJ x mol(-1), respectively. Using capillary columns to generate saturated vapors has three major advantages: short equilibrium time, low consumption of sample, and safe handling of toxic compounds. PMID- 11403280 TI - Evaluation of three-dimensional microchannel glass biochips for multiplexed nucleic acid fluorescence hybridization assays. AB - Three-dimensional, flow-through microchannel glass substrates have a potential for enhanced performance, including increased sensitivity and dynamic range, over traditional planar substrates used in medium-density microarray platforms. This paper presents a methodology for the implementation of multiplexed nucleic acid hybridization fluorescence assays on microchannel glass substrates. Fluorescence detection was achieved, in a first instance, using conventional low-magnification microscope objective lenses, as imaging optics whose depth-of-field characteristics match the thickness of the microchannel glass chip. The optical properties of microchannel glass were shown, through experimental results and simulations, to be compatible with the quantitative detection of heterogeneous hybridization events taking place along the microchannel sidewalls, with detection limits for oligonucleotide targets in the low-attomole range. PMID- 11403281 TI - Time-resolved total internal reflection fluorometry study on polarity at a liquid/liquid interface. AB - The polarity of a water/oil (oil: cyclohexane, carbon tetrachloride, toluene, chlorobenzene, o-dichlorobenzene, or 1,2-dichloroethane) interface was investigated by means of time-resolved total-internal-reflection (TIR) fluorescence spectroscopy of a polarity-sensitive probe: sulforhodamine B (SRB). In bulk solutions, the nonradiative decay rate constant of SRB increased with an increase in a solvent polarity parameter [ET(30)], and this relationship was used to estimate the polarities of water/oil interfaces. For the oil having a relatively low solvent polarity [ET(30) < 35 kcal/mol], the polarity of the water/ oil interface agreed with that of the arithmetic average of the polarities of the two phases [ET(30)calc]. For water/odichlorobenzene and water/1,2 dichloroethane interfaces [ET(30) of the oil > 35 kcal/mol], on the other hand, the interfacial polarity determined by TIR spectroscopy was lower than the ET(30)calc. The results are discussed in terms of thickness/roughness of the water/oil interface. PMID- 11403282 TI - Solid-phase microextraction using fused-silica fibers coated with sol-gel-derived hydroxy-crown ether. AB - A novel solid-phase microextraction (SPME) fiber containing hydroxydibenzo-14 crown-4 (OH-DB14C4)/hydroxy-terminated silicone oil (OH-TSO) was first prepared by a sol-gel method and investigated for the determination of phenols. The possible mechanism is discussed and confirmed by IR spectra. The coating has stable performance in high temperature (to 350 degrees C) and solvents (organic and inorganic) due to the chemical binding between the coating and the fiber surface. The addition of crown ether enhances the polarity of the coating compared with that of the sol-gel OH-terminated silicone oil fiber and, accordingly, provides higher extraction efficiency for polar phenolic compounds. On the other hand, OH-terminated silicone oil in the coating can not only increase the length of network but also help to spread the stationary phase on the silica surface uniformly. The fluorescence microscopy experiment suggests the benefit the more uniform surface of the sol-gel-derived OHDB14C4/OH-TSO fiber in comparison with sol-gelderived OH-DB14C4 fiber. Some parameters of the SPME fiber for the determination of phenols were investigated. Limits of detection of the phenols are below 1.0 ng/mL, and the precisions are from 2.9 to 4.6% (n = 6). Linear ranges were found to be 0.1-10 microg/mL The sensitivity of the method is enhanced at a low-pH level (pH approximately 1) and with the addition of salt. The method was applied to the analysis of wastewater sample from a paper mill. PMID- 11403283 TI - Evaluation of a multidimensional solid-phase extraction platform for highly selective on-line cleanup and high-throughput LC-MS analysis of triazines in river water samples using molecularly imprinted polymers. AB - A novel highly selective sample cleanup procedure based on the use of molecularly imprinted polymers (MIPs) as solid-phase extraction materials has been evaluated with respect to its applicability and routine use in environmental analysis. The method comprises the combination of a restricted access material (RAM) and a MIP allowing a selective sample preparation to be achieved in the online mode. This combination is called the size-selective sample separation and solvent switch (six-SPE). The RAM column combines size exclusion and adsorption chromatography, reducing the concentration of matrix molecules by a cutoff of 15 kDa. The MIP column selectively retains the triazine analytes whereas the residual matrix is not retained and separated completely. Thus, the automated RAM-MIP is capable of excluding all matrix and nontarget compounds. The cleaned and enriched extract is subsequently eluted to an HPLC column and analyzed by LC-MS. A complete on-line analysis cycle including multidimensional solid-phase extraction, separation, and detection takes less than 15 min. Terbuthylazine, atrazine, propazine, simazine, ametryn, prometryn, irgarol, and also the metabolites deethylatrazine and deisopropylatrazine can be determined without any matrix interferences, e.g., by humic acids. The whole setup is fully automated and may be continuously operated. Nonspecific interactions with the polymer are below 1% in all cases. The accuracy of the LC-MIP-LC-MS system was controlled using a certified reference material (Aquacheck). The applicability of the method to the cleanup of real samples was demonstrated by injection of contaminated river water samples. The stability of different polymers was tested by consecutive injections, and it was shown that the performance of the materials did not vary even after more than 300 enrichment and desorption cycles. PMID- 11403284 TI - Stable isotope ratio measurements on highly enriched water samples by means of laser spectrometry. AB - We demonstrate the feasibility of using laser spectrometry (LS) to analyze isotopically highly enriched water samples (i.e., delta2H < or = 15,000/1,000, 18O < or = 1,200/1,000), as often used in the biomedical doubly labeled water (DLW) method to quantify energy metabolism. The method is an important extension of a recently developed infrared laser direct absorption spectrometer. The measurements on highly enriched, small-size (10 microL liquid water) samples show a clearly better accuracy for the 2H/1H ratio. In the case of 18O/16O, the same level of accuracy is obtained as with conventional isotope ratio mass spectrometer (IRMS) analysis. With LS, the precision is better for both 18O/ 16O and 2H/1H. New is the ability to measure 17O/16O with the same accuracy as 18O/16O. A major advantage of the present technique is the absence of chemical sample preparation. The method is proven to be reliable and accurate and is ready to be used in many biomedical applications. PMID- 11403285 TI - An extraction chromatography method for Hf separation prior to isotopic analysis using multiple collection ICP-mass spectrometry. AB - A novel method for the single-step separation of Zr + Hf from all matrix elements of geological samples has been developed for Hf isotopic measurements using multiple collector-ICP-mass spectrometry. The method combines an effective sample decomposition by LiBO2 fusion with a selective separation of Hf + Zr by a solid phase extraction material based on dipentyl pentyl phosphonate, commercially available as U-TEVA.Spec. Using this simple and rapid procedure, Hf and Zr can be isolated in a single separation step with good recoveries (>90%) and satisfactory blank levels (approximately 55 pg of Hf), so that a subsequent isotopic measurement with ICPMS is possible. An excellent separation from rock-forming constituents is achieved, including those elements (Al, P, Ti, Cr, Fe, Mo, etc.) known to interfere in conventional separation methods based on ion-exchange techniques. The potential of this new method for Hf isotopic analysis is demonstrated by replicate MC-ICPMS measurements of 176Hf/177Hf ratios in seven international reference materials of silicate rocks, spanning a range of Hf contents and bulk compositions. PMID- 11403286 TI - Immunochemical approaches for purification and detection of TNT traces by antibodies entrapped in a sol-gel matrix. AB - A highly sensitive immunochemical method for immunoaffinity purification (IAP) and detection of trace amounts of TNT was developed on the basis of antibodies (Abs) in a ceramic matrix (sol-gel). The study resulted in: (i) a highly sensitive and reproducible TNT ELISA (I50 and I20 values of 0.4 +/- 0.09 ppb and 0.12 +/- 0.03 ppb, respectively; n = 12), which is highly specific to TNT; and (ii) successful entrapment of the Abs that bound free analyte from solution. Binding was found to be highly reproducible, dose dependent, and only slightly (1.2-1.8-fold) lower than that in solution. The entrapped Abs did not leach from the matrix and were tolerant of absolute ethanol, acetone, and acetonitrile. Bound analytes could be easily eluted from the sol-gel matrix at high recoveries. The sol-gel-based IAP method described above introduces a simple one-step procedure that has a high potential to serve as a suitable and convenient immunochromatographic device for cleanup and concentration of TNT from "real field" samples in a manner that complies with both chemical and immunochemical residue analysis methods. PMID- 11403287 TI - Determination of diffusion coefficients of electroactive species in time-of flight experiments using a microdispenser and microelectrodes. AB - Two novel methods for the determination of diffusion coefficients of redox species combining the special properties of microdispensing devices and microelectrodes are presented. Both are based on the local application of tiny volumes of the redox-active species by means of a dispenser nozzle at a defined distance from the surface of a microelectrode. The microelectrode, which is inserted through the bottom into an electrochemical cell, is held at a constant potential sufficient to oxidize or reduce the electro-active species under diffusional control. The dispenser, which is filled with the electro-active species, can be positioned by means of micrometer screws over the microelectrode. After dispensing a defined number of droplets near the microelectrode surface, the current through the microelectrode is recorded, usually yielding a peak shaped curve having a defined time delay between the shooting of the droplets and the maximum current. The time that is necessary to attain maximum current, together with the known distance between two dispensing points, can be used to determine the diffusion coefficient of the electroactive species without knowledge of any system parameters, such as concentration of the redox species, diameter of the electroactive surface or number of transferred electrons. A similar method for the determination of diffusion coefficient of redox species involves a second redox species for calibration purposes. A mixture of both species is shot close to the microelectrode surface. Due to the different formal potentials of the redox species that are used, they can be distinguished in sequential experiments by variation of the potentials that are applied to the microelectrode, and it is thus possible to determine the individual transit times of the redox species independently. The difference in the transit times, together with the known diffusion coefficient of one of the redox species, can be used to calculate the unknown diffusion coefficient of the second one. PMID- 11403288 TI - A factorial analysis of silanization conditions for the immobilization of oligonucleotides on glass surfaces. AB - The modification of glass surfaces with (3-mercaptopropyl)trimethoxysilane and the application of this to DNA chip technology are described. A range of factors influencing the silanization method, and hence the number of surface-bound, chemically active thiol groups, were investigated using a design of experiment approach based on analysis of variance. The number of thiol groups introduced on glass substrates were measured directly using a specific radiolabel, [14C]cysteamine hydrochloride. For liquid-phase silanization, the number of surface-bound thiol groups was found to be dependent on both postsilanization thermal curing and silanization time and relatively independent of silane concentration, reaction temperature, and sample pretreatment. Depending on the conditions used in liquid-phase silanization, (1.3-9.0) x 10(12) thiol groups/cm2 on the glass samples were bound. The reliability and repeatability of liquid- and vacuum-phase silanization were also investigated. Eighteen-base oligonucleotide probes were covalently attached to the modified surfaces via a 3'-amino modification on the DNA and subsequent reaction with the cross-linking reagent N (gamma-maleimidobutyryloxy) succinimide ester (GMBS). The resulting probe levels were determined and found to be stoichiometric with that of the introduced thiol groups. These results demonstrate that silanization of glass surfaces under specific conditions, prior to probe attachment, is of great importance in the development of DNA chips that use the simple concept of the covalent attachment of presynthesized oligonucleotides to silicon oxide surfaces. PMID- 11403289 TI - Fabrication and characterization of microwell array chemical sensors. AB - Microwell arrays have been fabricated on the distal face of coherent fiber-optic bundles. A typical microwell array comprises approximately 3,000 individual optical fibers that were etched chemically. Individual microwells were 1 to 14 microm deep with approximately 22-microm widths and were filled partially with a chemical sensing (polymer + dye) layer to produce a microwell array sensor (MWAS). MWASs were fabricated using a technically expedient, photoinitiated polymerization reaction whereby a approximately 2 to 10-microm thick pH-sensitive or O2-sensitive sensing layer was immobilized inside each microwell. The pH sensing layer comprised fluorescein isothiocyanate-dextran conjugate immobilized in a photopolymerizable poly(vinyl alcohol) membrane. The O2-sensing layer comprised a ruthenium metal complex entrapped in a gas-permeable photopolymerizable siloxane membrane. pH and PO2 were quantitated by acquiring luminescence images using an epifluorescence microscope/charge-coupled device imaging system. The pH-sensitive MWAS displayed a pKa of approximately 6.4 and a response time of approximately 2.5 s. The O2-sensitive MWAS behaved according to a nonlinear Stern-Volmer model with a maximum I0/I of approximately 4 and a response time of approximately 2.5 s. MWASs are advantageous in that suitably sized samples such as single biological cells can be co-localized with the sensing matrix in individual microwells. PMID- 11403290 TI - Integrating an ultramicroelectrode in an AFM cantilever: combined technology for enhanced information. AB - We present a novel approach to develop and process a microelectrode integrated in a standard AFM tip. The presented fabrication process allows the integration of an electroactive area at an exactly defined distance above of the end of a scanning probe tip and the subsequent remodeling and sharpening of the original AFM tip using a focused ion beam (FIB) technique (See ref 1 for patent information). Thus, the functionality of scanning electrochemical microscopy (SECM) can be integrated into any standard atomic force microscope (AFM). With the demonstrated approach, a precisely defined and constant distance between the microelectrode and the sample surface can be obtained, alternatively to the indirect determination of this distance usually applied in SECM experiments. Hence, a complete separation of the topographical information and the electrochemical signal is possible. The presented technique is a significant step toward electrochemical imaging with submicrometer electrodes as demonstrated by the development of the first integrated frame submicroelectrode. PMID- 11403291 TI - Optical multibead arrays for simple and complex odor discrimination. AB - A fiber optic bead-based sensor array platform has been employed to discriminate between six different odors and air carrier gas. Six different bead sensor types, with over 250 replicates of each, were monitored before, during, and after odor exposure to produce time-dependent fluorescence response patterns that were unique for each sensor-analyte combination. A total of 2,683 sensors were analyzed with respect to changes in their fluorescence, and signals from identical sensor beads were averaged to improve signal-to-noise ratios. Analyte classification rates of 100% were achieved for three complex (coffee bean) odors and three pure (simple) odors (toluene, acetone, 1,3-dinitrotoluene) measured at their highest relative concentrations. When lower odor concentrations were employed, the system exhibited better than 85% classification rates for analyte discrimination. Sensor response repeatability to these odor stimuli has also been quantified statistically, which is vital in defining the detection limit of the overall system. These results demonstrate, for the first time, the utility of our bead array technology for discriminating between different odor types at various dilution levels. PMID- 11403292 TI - Imaging of electroosmotic flow in plastic microchannels. AB - We have characterized electroosmotic flow in plastic microchannels using video imaging of caged fluorescent dye after it has been uncaged with a laser pulse. We studied flow in microchannels composed of a single material, poly(methyl methacrylate) (acrylic) or poly(dimethylsiloxane) (PDMS), as well as in hybrid microchannels composed of both materials. Plastic microchannels used in this study were fabricated by imprinting or molding using a micromachined silicon template as the stamping tool. We examined the dispersion of the uncaged dye in the plastic microchannels and compared it with results obtained in a fused-silica capillary. For PDMS microchannels, it was possible to achieve dispersion similar to that found in fused silica. For the acrylic and hybrid microchannels, we found increased dispersion due to the nonuniformity of surface charge density at the walls of the channels. In all cases, however, electroosmotic flow resulted in significantly less sample dispersion than pressure-driven flow at a similar velocity. PMID- 11403293 TI - Split luciferase as an optical probe for detecting protein-protein interactions in mammalian cells based on protein splicing. AB - We describe a new method for detecting protein-protein interactions in intact mammalian cells; the approach is based on protein splicing-induced complementation of rationally designed fragments of firefly luciferase. The protein splicing is a posttranslational protein modification through which inteins (internal proteins) are excised out from a precursor fusion protein, ligating the flanking exteins (external proteins) into a contiguous polypeptide. As the intein, naturally split DnaE from Synechocystis sp. PCC6803 was used: The N- and C-terminal DnaE, each fused respectively to N- and C-terminal fragments of split luciferase, were connected to proteins of interest. In this approach, protein-protein interactions trigger the folding of DnaE intein, wherein the protein splicing occurs and thereby the extein of ligated luciferase recovers its enzymatic activity. To test the applicability of this split luciferase complementation, we used insulin-induced interaction between known binding partners, phosphorylated insulin receptor substrate 1 (IRS-1) and its target N terminal SH2 domain of PI 3-kinase. Enzymatic luciferase activity triggered by insulin served to monitor the interaction between IRS-1 and the SH2 domain in an insulin dose-dependent manner, of which amount was assessed by the luminescent intensity. This provides a convenient method to study phosphorylation of any protein or interactions of integral membrane proteins, a class of molecules that has been difficult to study by existing biochemical or genetic methods. High throughput drug screening and quantitative analysis for a specific pathway in tyrosine phosphorylation of IRS-1 in insulin signaling are also made possible in this system. PMID- 11403294 TI - Determination of picomolar levels of iron in seawater using catalytic cathodic stripping voltammetry. AB - A new procedure for the direct determination of picomolar levels of iron in seawater is presented. Cathodic stripping voltammetry (CSV) is preceded by adsorptive accumulation of the iron(III)-2,3-dihydroxynaphthalene (DHN) complex from seawater, containing 20 microM DHN at pH 8.0, onto a static mercury drop electrode, followed by reduction of the adsorbed species. The reduction current is catalytically enhanced by the presence of 20 mM bromate. Optimized conditions include a 60-s adsorption period at -0.1 V and a voltammetric scan using sampled dc modulation at 10 Hz. In these conditions, a detection limit of 13 pM iron in seawater was achieved which can be lowered further by extending the adsorption time to 300 s. The new catalytic CSV method is approximately 5 times more sensitive than existing CSV methods and was tested on samples from the Atlantic Ocean. PMID- 11403295 TI - Construction and analytical characterization of Prussian-Blue-based carbon paste electrodes and their assembly as oxidase enzyme sensors. AB - This paper reports on the development and characterization of Prussian Blue modified carbon paste electrodes. New methods of matrix modification with Prussian Blue are reported. Two different carbon pastes have been prepared, using mineral oil or solid paraffin, thus obtaining different types of sensors whose behavior toward the electrochemical reduction of hydrogen peroxide has been fully characterized. Results obtained with Prussian Blue-modified electrodes showed a long operational lifetime, an excellent stability in a wide range of pH (3-9), a high sensitivity, and a fast response time. In addition, the coupling of solid carbon paste modified with Prussian Blue and the enzymes glucose oxidase and choline oxidase led to the assembling of biosensors that showed an optimum working range at alkaline pH. PMID- 11403296 TI - Development of rotating electrochemical detectors for capillary electrophoresis. AB - A rotating disk electrode (RDE) has been evaluated and optimized for the detection of electroactive species separated by capillary electrophoresis (CE). With catechol as a working model, the limit of detection was estimated to be 0.3 microM, i.e., approximately 2.5-fold better than that of the stationary disk electrode (0.7 microM). Separation efficiency was significantly improved as exemplified by an increase of theoretical plates from 26,000 plates/m at 0 rpm to 67,000 plates/m at 500 rpm. Of particular importance was the capability of RDE to alleviate electrode passivation and electrical interference associated with high separation potential fields. Therefore, rotation amperometry was especially useful for analytes such as phenolic compounds that tended to rapidly foul the electrode surface. The RDE/ CE system was capable of separation and determination of pentachlorophenol in contaminated soils, and the result obtained agreed well with conventional liquid chromatography, an EPA recommended procedure. PMID- 11403297 TI - Analysis of multiple samples using multiplex sample NMR: selective excitation and chemical shift imaging approaches. AB - Two improved approaches for the rapid analysis of multiple samples using multiplex sample NMR are described. In the first approach, frequency-selective 90 degrees radio frequency pulses and large pulsed field gradients are applied to excite and detect multiple samples in rapid succession. This method is advantageous for samples with relatively long longitudinal (T1) relaxation times. In the second approach, chemical shift imaging is applied to acquire both the spectral and spatial information of multiple samples simultaneously. Chemical shift imaging is more time-consuming than selective excitation; however, it is advantageous for detecting samples with short T1's and for signal averaging. Both approaches demonstrate the potential of multiplex sample NMR for carrying out high-throughput NMR detection. PMID- 11403298 TI - Determination of total sulfur at microgram per gram levels in geological materials by oxidation of sulfur into sulfate with in situ generation of bromine using isotope dilution high-resolution ICPMS. AB - We have developed a new, simple, and accurate method for the determination of total sulfur at microgram per gram levels in milligram-sized silicate materials with isotope dilution high-resolution inductively coupled plasma mass spectrometry equipped with a flow injection system. In this method, sulfur can be quantitatively oxidized by bromine into sulfate with achievement of isotope equilibrium between the sample and spike. Detection limits for 32S+ and 34S+ in the ideal solution and silicate samples were 1 and 6 ng mL(-1) and 0.07 and 0.3 microg g(-1), respectively. The total blank was 46 ng, so that a 40-mg silicate sample containing 10 mirog g(-1) sulfur can be measured with a blank correction of < 10%. This total blank can be lowered to 8 ng if a low-blank air system is used for evaporations. To evaluate the applicability of this method, we analyzed not only silicate reference materials with sulfur content of 5.25-489 microg g( 1) and sample sizes of 13-40 mg but also the Allende meteorite with a sulfur content of 2%. The reproducibility for various rock types was < 9%, even though blank corrections in some samples of low sulfur content were up to 24%. This method is suitable for analyzing geological samples as well environmental samples such as soils, sediments, and water samples. PMID- 11403299 TI - Use of ion mobility spectrometry to determine low levels of impurities in gases. AB - The use of ion mobility spectrometry (IMS) for the determination of trace moisture and oxygen in bulk nitrogen has been explored. IMS utilizes atmospheric pressure ionization to ionize trace impurities in the sample gas. Mobility differences between trace impurity ions are exploited to separate these ions. Our results indicate that an IMS can indeed be used to detect < 1 ppb O2 and H2O in bulk nitrogen. Due to the nature of the interaction between trace moisture and oxygen, a multivariate calibration has to be used to obtain quantitative results, even at levels below 2 ppb. PMID- 11403300 TI - Identification of individual proteins in complex protein mixtures by high resolution, high-mass-accuracy MALDI TOF-mass spectrometry analysis of in solution thermal denaturation/enzymatic digestion. AB - Identification of individual proteins in complex protein mixtures by high resolution (HR), high-mass-accuracy matrix-assisted laser desorption/ionization (MALDI) time-of-flight mass spectrometry (TOF-MS) is demonstrated for synthetic protein mixtures. Instead of chemical denaturation, thermal denaturation followed by in-solution trypsin digestion is used to achieve uniform digestion of the constituents of the protein mixture. Protein identification is carried out using protein database searches with search scoring systems, which seems more effective than conventional peptide mass mapping without using a scoring system. Identification of individual proteins by MALDI HR-TOF-MS peptide mass mapping dramatically reduces data acquisition/analysis time and does not require special equipment for sample preparation/transfer prior to mass spectral analysis. PMID- 11403301 TI - Dynamics of iron release from transferrin N-lobe studied by electrospray ionization mass spectrometry. AB - Transferrins constitute a class of metalloproteins that are involved in circulatory iron transport in a variety of species. The metal ion-binding properties of these proteins have been the focus of extensive research efforts in the past decade due to their extreme importance in a variety of biological and healthcare-related fields. The large size of these proteins, as well as the presence of high-spin metal ions (e.g., Fe3+), limits the use of NMR. In this work, we report on the use of electrospray ionization mass spectrometry (ESI MS) to study dynamics of the transferrin system in vitro under conditions that are designed to mimic the endosomal environment. ESI MS is shown to provide valuable insights into the mechanistic aspects of metal ion-binding/release by transferrins and is complementary to other spectroscopic techniques. Conformational stability of the complex is evaluated based on the appearance of the charge-state distribution of protein ions, while the composition of the protein-ligand complex is determined based on the mass of the protein ions. In the absence of iron chelators, a stepwise dissociation of the ternary complex (protein-metal ion-synergistic anion) is observed as the solution pH is gradually decreased. Although the release of synergistic anion from the complex is initiated at typical endosomal pH levels (i.e., 5.5), metal ion remains largely bound to the protein until the pH is lowered to a level of approximately 4.5. Under these conditions, a significant fraction of the protein populates unfolded conformations. In stark contrast to this behavior, addition of an iron chelating agent (citrate) to the protein solution results in facile iron release at typical endosomal pH levels without any detectable unfolding of the protein. The mass spectral data lends further credibility to the notion that the holoprotein samples conformations that are specific to the apo form (e.g., "open conformation"), from which iron dissociation most likely occurs. The results of the present study demonstrate that ESI MS can be used to model metal ion release from transferrin under conditions that are designed to mimic the physiological environment. PMID- 11403302 TI - A photometric screening method for dimeric naphthylisoquinoline alkaloids and complete on-line structural elucidation of a dimer in crude plant extracts, by the LC-MS/LC-NMR/LC-CD triad. AB - An efficient evaluation procedure for the chemical screening and on-line structural elucidation of dimeric naphthylisoquinoline alkaloids has been developed. The method is based on the lead tetraacetate oxidation of the central binaphthalene core of the alkaloids. UV spectra of the extracts after addition of the oxidant show, in the presence of naphthylisoquinoline dimers, the appearance of a characteristic long-wavelength absorption indicative of dinaphthoquinones. The efficiency and relevance of the method has been demonstrated in the discovery of a constitutionally and configurationally new dimeric naphthylisoquinoline alkaloid, named ancistrogriffithine A (4), from the previously uninvestigated Asian liana Ancistrocladus griffithii. After verification of this screening result by LC-ESI-MS/MS, the constitution and the relative configuration of the compound were elucidated on line, by LC-NMR and LC-CD on the extract. Using an LC NMR-WET-ROESY experiment, itwas possible for the first time to determine the relative axial configuration of a natural biaryl compound on line, by observing long-range ROE interactions. Finally, an oxidative degradation right on the extract delivered the absolute configuration of 4, without isolation of the alkaloid. Ancistrogriffithine A is the as yet only dimeric naphthylisoquinoline from an Asian Ancistrocladaceae plant. PMID- 11403303 TI - Phosphoprotein isotope-coded affinity tag approach for isolating and quantitating phosphopeptides in proteome-wide analyses. AB - A method has been developed that utilizes phosphoprotein isotope-coded affinity tags (PhIAT) that combines stable isotope and biotin labeling to enrich and quantitatively measure differences in the O-phosphorylation states of proteins. The PhIAT labeling approach involves hydroxide ion-mediated beta-elimination of the O-phosphate moiety and the addition of 1,2-ethanedithiol containing either four alkyl hydrogens (EDT-D0) or four alkyl deuteriums (EDT-D4) followed by biotinylation of the EDT-D0/D4 moiety using (+)-biotinyl-iodoacetamidyl-3,6 dioxaoctanediamine. The PhIAT reagent, which contains the nucleophilic sulfhydryl and isotopic label covalently linked to a biotin moiety, was synthesized and has the potential utility to reduce the O-phosphorylation derivatization into a one step process. The PhIAT labeling approach was initially demonstrated using the model phosphoprotein beta-casein. After proteolytic digestion, the PhIAT-labeled peptides were affinity isolated using immobilized avidin and analyzed using capillary reversed-phase liquid chromatography-mass spectrometry. PhIAT-labeled beta-casein peptides corresponding to peptides containing known sites of O phosphorylation were isolated and identified. The PhIAT labeling method was also applied to a yeast protein extract. The PhIAT labeling technique provides a reliable method for making quantitative measurements of differences in the O phosphorylation state of proteins. PMID- 11403304 TI - On-line coupling of capillary gel electrophoresis with electrospray mass spectrometry for oligonucleotide analysis. AB - Homooligodeoxyribonucleotides differing one nucleotide in length from 12- to 15 mer and from 17- to 20-mer were separated by size with capillary gel electrophoresis (CGE) using an entangled polymer solution in coated capillaries. The resolved components were analyzed by on-line coupling of CGE with electrospray mass spectrometry (ES-MS), denoted as CGE/ES-MS, in the full-scan negative ion detection mode. Baseline separation was achieved for the 12-15-mer oligonucleotide mixtures. Both synthetic phosphodiester oligonucleotide mixtures as well as their phosphorothioate analogues, serving as model compounds for antisense oligonucleotides, could be analyzed by on-line CGE/ES-MS coupling. Terminally phosphorylated and nonphosphorylated synthetic failure sequences could be electrophoretically separated and mass spectrometically characterized as well. This methodology might be a useful tool for synthesis control of phosphodiester oligonucleotides as well as for analysis of phosphorothioate analogues as they are used in antisense drug development. PMID- 11403305 TI - Implementation and uses of automated de novo peptide sequencing by tandem mass spectrometry. AB - There are several computer programs that can match peptide tandem mass spectrometry data to their exactly corresponding database sequences, and in most protein identification projects, these programs are utilized in the early stages of data interpretation. However, situations frequently arise where tandem mass spectral data cannot be correlated with any database sequences. In these cases, the unmatched data could be due to peptides derived from novel proteins, allelic or species-derived variants of known proteins, or posttranslational or chemical modifications. Two additional problems are frequently encountered in high throughput protein identification. First, it is difficult to quickly sift through large amounts of data to identify those spectra that, due to poor signal or contaminants, can be ignored. Second, it is important to find incorrect database matches (false positives). We have chosen to address these difficulties by performing automatic de novo sequencing using a computer program called Lutefisk. Sequence candidates obtained are used as input in a homology-based database search program called CIDentify to identify variants of known proteins. Comparison of database-derived sequences with de novo sequences allows for electronic validation of database matches even if the latter are not completely correct. Modifications to the original Lutefisk program have been implemented to handle data obtained from triple quadrupole, ion trap, and quadrupole/time-of flight hybrid (Qtof) mass spectrometers. For example, the linearity of mass errors due to temperature-dependent expansion of the flight tube in a Qtof was exploited such that isobaric amino acids (glutamine/lysine and oxidized methionine/ phenylalanine) can be differentiated without careful attention to mass calibration. PMID- 11403306 TI - Accurate mass liquid chromatography/mass spectrometry on quadrupole orthogonal acceleration time-of-flight mass analyzers using switching between separate sample and reference sprays. 2. Applications using the dual-electrospray ion source. AB - A new electrospray dual sprayer, LockSpray, was developed for accurate mass measurements on a quadrupole orthogonal acceleration time-of-flight mass spectrometer (oa-Q-ToF). With the dual-sprayer ion source, both sprays are orthogonal to each other. A mechanism similar to the one employed on the multiplexed electrospray source (MUX) allows switching between reference and sample sprayer. The reference sprayer is optimized for low flow rates, whereas the sample sprayer is a conventional Z-spray type sprayer. Earlier work using a modified MUX ion source on an orthogonal acceleration time-of-flight instrument showed promising results. In this paper, examples obtained with the LockSpray, specifically designed for accurate mass measurements on an oa-Q-ToF, are presented. The examples include results obtained for the identification of impurities in drug substances such as cimetidine and rosiglitazone, using accurate mass tandem mass spectrometry in both positive and negative ion electrospray modes. Good mass accuracies, i.e., within 2 mDa of the theoretical value, were obtained in MS and MS/MS operation. PMID- 11403307 TI - Solvent extraction, ion chromatography, and mass spectrometry of molybdenum isotopes. AB - A procedure was developed that allows precise determination of molybdenum isotope abundances in natural samples. Purification of molybdenum was first achieved by solvent extraction using di(2-ethylhexyl) phosphate. Further separation of molybdenum from isobar nuclides was obtained by ion chromatography using AG1-X8 strongly basic anion exchanger. Finally, molybdenum isotopic composition was measured using a multiple collector inductively coupled plasma hexapole mass spectrometer. The abundances of molybdenum isotopes 92, 94, 95, 96, 97, 98, and 100 are 14.8428(510), 9.2498(157), 15.9303(133), 16.6787(37), 9.5534(83), 24.1346(394), and 9.6104(312) respectively, resulting in an atomic mass of 95.9304(45). After internal normalization for mass fractionation, no variation of the molybdenum isotopic composition is observed among terrestrial samples within a relative precision on the order of 0.00001-0.0001. This demonstrates the reliability of the method, which can be applied to searching for possible isotopic anomalies and mass fractionation. PMID- 11403308 TI - Improving spot homogeneity by using polymer substrates in matrix-assisted laser desorption/ionization mass spectrometry of oligonucleotides. AB - We describe a method for improving the homogeneity of MALDI samples prepared for analysis of small, single-stranded oligonucleotides using the widely used DNA matrix system, 3-hydroxypicolinic acid/picolinic acid/ ammonium citrate. This matrix system typically produces large crystals around the rim of the dried sample and requires tedious searching of this rim with the laser. However, when a substrate is prepared using both Nafion and a hydrophilic, high-molecular-weight polymer, such as linear polyacrylamide, linear poly(ethylene oxide), or methyl cellulose, oligonucleotide-doped matrix crystals tend to be smaller and more uniformly distributed across the entire spot, thus decreasing the time that is required for locating a usable signal. In addition to MALDI characterization of the spatial distribution of "sweet spots," fluorescence microscopy allows for imaging dye-labeled DNA in dried MALDI spots. The mechanism of enhanced uniformity may involve increased viscosity in the MALDI sample droplet due to partial solubilization of the substrate by the MALDI sample solvent as well as partitioning of the matrix or DNA between the solvent and the undissolved portion of the polymer substrate. PMID- 11403309 TI - Stereoselective determination of amino acids in beta-amyloid peptides and senile plaques. AB - A novel method for the determination of the enantiomeric composition of peptides is presented. In this paper, the focus has been on beta-amyloid peptides from deceased Alzheimer's disease patients. The peptides are hydrolyzed using mineral acid. The free amino acids are derivatized with the chiral reagent (+)- or (-)-1 (9-anthryl)-2-propyl chloroformate and subsequently separated using micellar electrokinetic chromatography (MEKC) and detected using laser-induced fluorescence (LIF) detection. The high separation efficiency of the MEKC-LIF system, yielding approximately 1 million theoretical plates/m for most amino acids, facilitates the simultaneous chiral determination of nine amino acids. The samples that have been analyzed were standard 1-40 beta-amyloid peptides, in vitro precipitated beta-amyloid fibrils, and human senile plaque samples. PMID- 11403310 TI - Extended thermodynamic approach to ion interaction chromatography. AB - The chromatographic behavior of charged analytes in ion interaction chromatography (IIC) is theoretically investigated. The chemical modifications of the stationary and mobile phases in the presence of ion interaction reagent (IIR) are theoretically shown to change the partition coefficient for charged molecules. The most reliable literature experimental results concerning retention behavior of charged molecules in IIC were used to test the new theory. Retention equations are compared with those that can be obtained from the most important retention models in IIC. The present exhaustive retention model, which is well founded in physical chemistry, goes further than the previous ones whose retention equations can be viewed as limiting cases of the present theory. The present extended thermodynamic approach reduces to stoichiometric or electrostatic retention models if the surface potential or pairing equilibria are respectively neglected. Moreover, it is able to quantitatively explain experimental evidences that cannot be rationalized by the existing retention models. PMID- 11403311 TI - Capillary electrophoresis of sialic acid-containing glycoprotein. Effect of the heterogeneity of carbohydrate chains on glycoform separation using an alpha1-acid glycoprotein as a model. AB - alpha1-Acid glycoprotein (AGP) showed multiple peaks on separation using capillary electrophoresis in a chemically modified capillary with dimethylpolysiloxane at slightly acidic conditions. We analyzed glycoforms of AGP species after separation by ion-exchange chromatography, Con A affinity chromatography, and Cu(II)-chelating affinity chromatography. The AGP species thus obtained were digested with N-glycosidase F, and the released carbohydrate chains were analyzed by high-performance liquid chromatography after labeling with 3-aminobenzoic acid. The results afforded basic information on the contribution of carbohydrate chains to the separation mechanism of glycoforms of AGP by capillary electrophoresis. In addition, we describe an easy method for AGP analysis in serum samples using the electrokinetic injection. PMID- 11403312 TI - Integrated microfluidic system enabling protein digestion, peptide separation, and protein identification. AB - An integrated platform is presented for rapid and sensitive protein identification by on-line protein digestion and analysis of digested proteins using electrospray ionization mass spectrometry or transient capillary isotachophoresis/capillary zone electrophoresis with mass spectrometry detection. A miniaturized membrane reactor is constructed by fabricating the microfluidic channels on a poly(dimethylsiloxane) substrate and coupling the microfluidics to a poly(vinylidene fluoride) porous membrane with the adsorbed trypsin. On the basis of he large surface area-to-volume ratio of porous membrane media, adsorbed trypsin onto the poly(vinylidene fluoride) membrane is employed for achieving ultrahigh catalytic turnover. The extent of protein digestion in a miniaturized membrane reactor can be directly controlled by the residence time of protein analytes inside the trypsin-adsorbed membrane, the reaction temperature, and the protein concentration. The resulting peptide mixtures can either be directly analyzed using electrospray ionization mass spectrometry or further concentrated and resolved by electrophoretic separations prior to the mass spectrometric analysis. This microfluidic system enables rapid identification of proteins in minutes instead of hours, consumes very little sample (nanogram or less), and provides on-line interface with upstream protein separation schemes for the analysis of complex protein mixtures such as cell lysates. PMID- 11403313 TI - Narrow sample channel injectors for capillary electrophoresis on microchips. AB - In microchip CE, sample injection is generally achieved through cross, double tee, or tee injector structures. In these reported approaches, channel width and depth are uniform at the injection intersection. Here, we present cross and tee injectors having narrow sample channels. Using a cross injector with reduced sample channel width, resolution, column efficiency, and sensitivity are remarkably improved. Furthermore, no leakage control is required in both injection and separation phases, making the microchip CE system more user friendly. Good resolution can also be obtained using tee injectors with narrow sample channels, which would otherwise be impossible using conventional tee injectors. Using the narrow sample channel tee injector instead of conventional cross and double-tee injectors, the number of reservoirs in multiplexed systems can be reduced to N + 2 (N, the number of paralleled CE systems), the real theoretical limit. The virtues of the novel injectors were demonstrated with poly(dimethylsiloxane)-glass chips incorporating eight parallel CE channels. PMID- 11403314 TI - Sensitive and universal indirect chemiluminescence detection for capillary electrophoresis of cations using cobalt(II) as a probe ion. AB - Highly sensitive and universal indirect chemiluminescence detection for capillary electrophoresis of cations was described. This novel method is based on use of the ultrasensitive cobalt(II) as a probe ion in the running buffer. A strong and stable background chemiluminescent signal can be generated by the luminol hydrogen peroxide reaction catalyzed by cobalt(II) ion. Displacement of the cobalt(II) probe ion in the running buffer by a migrating sample cation results in a quantifiable decrease in the background signal. The conditions for electrophoresis and the chemiluminescent reaction were systematically investigated using a commercial capillary electrophoresis instrument with an in house-built chemiluminescence detector. Under the optimal conditions, the detection limits of the concentration for manganese(II), cadmium(II), nickel(II), lead(II), and 14 lanthanides were (3.0-6.0) x 10(-9) mol/L (S/N = 3), which was approximately 3 orders of magnitude better than indirect UV detection and 2 orders better than indirect laser-induced fluorescent detection. A mixture of 18 metal ions including 14 lanthanides was efficiently separated within 3.5 min using lactate to partially complex the metal ions. Our data demonstrated that CE with indirect CL detection was a powerful and universal tool for analysis of inorganic and organic cations. PMID- 11403315 TI - Two-dimensional electrochromatography/capillary electrophoresis on a microchip. AB - A two-dimensional separation system on a microfabricated device was demonstrated using open-channel electrochromatography as the first dimension and capillary electrophoresis as the second dimension. The first dimension was operated under isocratic conditions, and the effluent from the first dimension was repetitively injected into the second dimension every few seconds. A 25-cm separation channel with spiral geometry for open-channel electrochromatography was chemically modified with octadecylsilane and coupled to a 1.2-cm straight separation channel for capillary electrophoresis. Fluorescently labeled products from tryptic digests of beta-casein were analyzed in 13 min with this system. PMID- 11403316 TI - High-throughput microfabricated CE/ESI-MS: automated sampling from a microwell plate. AB - A new design for high-throughput microfabricated capillary electrophoresis/electrospray mass spectrometry (CE/ ESI-MS) with automated sampling from a microwell plate is presented. The approach combines a sample loading port, a separation channel, and a liquid junction, the latter for coupling the device to the MS with a miniaturized subatmospheric electrospray interface. The microdevice was attached to a polycarbonate manifold with external electrode reservoirs equipped for electrokinetic and pressure-fluid control. A computer-activated electropneumatic distributor was used for both sample loading from the microwell plate and washing of channels after each run. Removal of the electrodes and sample reservoirs from the microdevice structure significantly simplified the chip design and eliminated the need both for drilling access holes and for sample/buffer reservoirs. The external manifold also allowed the use of relatively large reservoirs that are necessary for extended time operation of the system. Initial results using this microfabricated system for the automated CE/ESI-MS analysis of peptides and protein digests are presented. PMID- 11403317 TI - Proteolysis in mixed organic-aqueous solvent systems: applications for peptide mass mapping using mass spectrometry. AB - The rate of protein digestion imposes significant limitations on high-throughput protein identification using mass spectrometry. In this report, we demonstrate that proteins are readily digested by trypsin in the presence of organic solvents such as methanol, acetone, 2-propanol, and acetonitrile. The rates of protein digestion in organic solvents, as indicated by the abundances of digest fragment ions in the mass spectrum, are increased relative to aqueous solution. In addition, amino acid coverage for the analyzed proteins increases in the presence of the organic solvents, and proteins that are resistant to proteolysis are readily digested. For example, a 68% amino acid sequence coverage was attained from a tryptic digest of myoglobin in < 5 min from an 80% acetonitrile solution, whereas no digest fragments were detected from a 5 min digestion in an aqueous solution. Moreover, the tryptic digestion of a complex protein mixture in an organic-aqueous solvent system showed significantly enhanced digestion for nearly all of the protein components. Enzymatic digestion in an organic-aqueous solvent system is a rapid, simple, and effective peptide mass-mapping technique. PMID- 11403318 TI - ECL for near-field imaging. PMID- 11403319 TI - Multipurpose nanopore sensors. PMID- 11403320 TI - Putting a fine point on ESI. PMID- 11403321 TI - Flexible immunoassays on a chip. PMID- 11403322 TI - Classifying enzyme inhibitors using MS. PMID- 11403323 TI - Microarray standards adopted. PMID- 11403324 TI - NACLA steps forward. PMID- 11403325 TI - Microfluidics: controlling fluids in small places. PMID- 11403326 TI - Prefractionation techniques in proteome analysis. PMID- 11403327 TI - DNA sequencers rely on CE. PMID- 11403328 TI - Teaching the essential principles. PMID- 11403329 TI - Alzheimer disease and associated disorders: project funding opportunities within the European community. AB - The European Union has funded numerous projects on Alzheimer disease and associated disorders (ADAD). In the funding years of 1995 and 1996, six million Euro were spent on ADAD projects. In 1996, 21% of applications were funded. Having had the opportunity to review all these projects, we offer insight into the funding procedure and some of our conclusions from reviewing all these projects. PMID- 11403330 TI - Environmental lead-210 and bismuth-210 accrue selectively in the brain proteins in Alzheimer disease and brain lipids in Parkinson disease. AB - We studied the occurrence of the environmental radon daughters, 210Po (alpha particles), and 210Bi (beta particles), in the protein and lipid fractions of cortical gray and subcortical white matter from the frontal and temporal lobes of human brains of persons with Alzheimer disease (AD), persons with Parkinson disease (PD), smokers, or persons with no previous evidence of clinical neurologic disease (controls). We found a 10-fold increase in 210Po and 210Pb radioactivity in the protein fraction from both the cortical gray and subcortical white matter in AD and smokers, and a similar increase in the lipid fraction in PD. The pathognomonic distribution of the radon daughters to the lipids in PD and to the proteins in AD was inferred to reflect the increase of local chlorine availability to which radon daughters bound selectively. Cigarette smoking strongly increases radon daughter retention in the central nervous system. PMID- 11403331 TI - The end of Alzheimer disease. PMID- 11403332 TI - Asia-Pacific Consensus Statement on dementia. AB - The 12 national Alzheimer's Associations that contributed to this Consensus Statement will continue to network with one another regionally and will continue to share relevant expertise and practical resources. It is expected that regional consensus on dementia will evolve in individual member countries and in the Asia Pacific region as a whole. It is the hope of the participating members of these two consensus conferences that this document will provide the inspiration, direction, and practical ideas to further advance the goals of national Alzheimer's Associations and to further advance dementia-related medical and service activities within the Asia-Pacific region. PMID- 11403333 TI - Asia regional meeting of the International Working Group for the Harmonization of Dementia Drug Guidelines: meeting report. AB - This meeting successfully catalyzed the establishment of a new working alliance between clinical dementia researches in Asia and identified common goals for the group to attain. The progress toward achieving these goals will be examined at the next Asia regional meeting, which is being planned for October 2002 in Beijing, China. This new regional working group will work with the IWG to overcome the existing methodological and regulatory obstacles impeding dementia treatment trials in Asia. PMID- 11403334 TI - Quality of Life in Dementia: state of the art--report of the International Working Group for Harmonization of Dementia Drug Guidelines and the Alzheimer's Society satellite meeting. PMID- 11403335 TI - Evidence that age-associated memory impairment is not a normal variant of aging. AB - The concept of age-associated memory impairment (AAMI) suggests that clinically recognized memory dysfunction can be a feature of normal aging. To determine whether AAMI represents a variant of normal aging, we longitudinally studied individuals meeting AAMI criteria for development of dementia. Two hundred two community-living individuals (mean age, 77 years) with or without mild memory impairment were assessed annually for an average of 3 years at the Washington University Alzheimer's Disease Research Center. At baseline, no individual was unequivocally demented, as defined by a Clinical Dementia Rating (CDR) score of 1 or greater. Modified National Institute of Mental Health criteria were used to identify individuals with AAMI who otherwise met a criterion for cognitive normality. The Short Blessed Test (SBT) was used as a measure of general cognitive function; conservative (SBT = 5) and permissive (SBT = 10) cutoff scores were used as indicators of cognitive normality. With the more permissive measure of cognitive normality, 59 (29%) of the 202 individuals met AAMI criteria. Progression to dementia by 3 years occurred in 42% of AAMI individuals versus 16% of the individuals who did not meet AAMI criteria. With the more restrictive SBT cutoff of 5, 22% of individuals met AAMI criteria; progression to dementia occurred in 31% of these individuals versus 9% of the individuals without AAMI. Survival times to dementia differed significantly between AAMI and non-AAMI groups defined by either cutoff score. Our findings indicate that individuals with AAMI have a three-fold greater risk for development of dementia than individuals who do not meet AAMI criteria. Hence, AAMI may represent a dementia prodrome rather than a benign variant of aging. PMID- 11403336 TI - Pilot tolerability studies of hydroxychloroquine and colchicine in Alzheimer disease. AB - Anti-inflammatory drugs may be useful in the treatment of Alzheimer disease (AD). In preparation for therapeutic trials, we conducted pilot feasibility studies of hydroxychloroquine alone and in combination with colchicine in subjects with AD. A total of 20 subjects with probable AD were treated with hydroxychloroquine 200 mg twice daily for 11 weeks, or hydroxychloroquine 200 mg twice daily plus colchicine 0.6 mg twice daily for 12 weeks; subjects were monitored for adverse medical, cognitive, or behavioral effects. Neither regimen caused adverse effects on cognitive or behavioral assessment scores. There were no significant side effects in subjects receiving hydroxychloroquine alone; 2 subjects receiving the two drugs together experienced diarrhea. We conclude that these regimens of anti inflammatory therapy are well tolerated in subjects with AD, indicating the feasibility of large-scale therapeutic trials of these agents. PMID- 11403337 TI - Self-control and cooperation: different concepts, similar decisions? A question of the right perspective. AB - Conflicts between an individual's long-term and short-term goals (i.e., self control conflicts) and conflicts between individual and collective interests (mixed-motive situations) share some interesting features. In this article both types of conflicts are approached from a perspective hypothesis. The hypothesis holds that a decision maker's perspective on his or her decision determines whether either the long-term goals (the collective interests) or the short-term goals (the individual interests) will guide behavior. The hypothesis also implies that factors known to enhance cooperative choices in mixed-motive situations should also facilitate self-control. Three such factors are evaluated: group identification, self-efficacy, and mutual trust. We conclude that the perspective in which choice is considered part of an identifiable series of behaviors is most likely to result in cooperation or self-control. PMID- 11403338 TI - The effects of frequency of social interaction and relationship closeness on reward allocation. AB - In this study of how the frequency of interaction affects decisions on reward allocation, 320 Chinese adults were randomly assigned to 8 conditions and instructed to read a scenario describing 2 co-workers completing a task and obtaining a bonus. The 8 conditions differed in the type of relationships, the frequency of past interaction between co-workers, and the possibility of future interaction. Each participant was instructed to act as the allocator and to hypothetically allocate a monetary reward to the co-worker. The results indicated that both past interaction and future interaction between the co-workers significantly influenced the allocation decision. These results also suggested that the frequency of 2 individuals' past interaction can be predictive of the closeness of their relationship. PMID- 11403339 TI - Normative perceptions in relation to substance use and HIV-risky sexual behaviors of college students. AB - In this study, 410 college students completed a survey assessing their personal substance use patterns and sexual behavior and their perceptions of other students' substance use and sexual behavior. Two parallel sets of questions were presented to reflect the two contextual conditions of being drunk or high versus being not drunk or high. Results indicated that number of recent sexual partners and normative perceptions of HIV-risky sexual behavior were the strongest predictors of personal HIV-risky behavior when not drunk or high. HIV-risky behavior when drunk or high was predicted by personal substance use as well as by number of recent partners and normative perceptions of peer sexual behavior. These findings suggest that HIV-prevention programs for college students should aim to correct overinflated perceptions of other students' high-risk sexual behavior. PMID- 11403340 TI - Engaging distortions: are we idealizing marriage? AB - The present study was an investigation of the premarital status of engagement in terms of relationship satisfaction and marital expectations using the Evaluation and Nurturing Relationship Issues, Communication and Happiness (ENRICH) Marital Satisfaction Scale (EMS) and its two subscales of Idealistic Distortion (ID) and Marital Satisfaction (MS) (D. G. Fournier, D. H. Olson, & J. M. Druckman, 1983). There were 104 students (23 men and 81 women), of which 15 were married, 19 were engaged, and 70 had extended dating relationships. On average, participants had been in the relationship for 3.8 years, and the mean age was 22 years. Results demonstrated that individuals engaged to be married had significantly higher idealistic distortion scores (M = 86.89) than did either married individuals (M = 56.67) or those in extended dating relationships (M = 61.19). Finally, a negative relation was found between length of relationships and marital satisfaction subscores. Results are discussed in light of factors contributing to such idealized thinking. PMID- 11403341 TI - Direct reports of motivation for task performance levels: some construct-related evidence. AB - The validity of direct self-reports of motivational force for task performance levels was examined in three studies. Such measures have been used as true scorelike criteria for computed measures of force and have advantages as measures in their own right. Results suggested that simple, direct self-reports should be useful in some research settings, but their validity as assessments of motivational force per se may be influenced by both wording and context. PMID- 11403342 TI - Concurrent validity of the Test of Nonverbal Intelligence in Parkinson's disease patients. AB - The Test of Nonverbal Intelligence (TONI-2; L. Brown, R. J. Sherbenou, & S. Johnsen, 1990) and Raven's Colored Progressive Matrices (RCPM; J. C. Raven, 1965) are defined as language-free measures of cognitive ability. The purpose of the present study was to investigate the relation between the RCPM and the TONI-2 for samples of patients with Parkinson's disease (n = 75) and controls (n = 47). A regression equation was computed to evaluate the relation of the RCPM scores to the TONI-2 quotient. Regression equation results indicate that there is a significant overlapping linear variance between the two measures in both patients and controls. PMID- 11403343 TI - Path model analyzed with ordinary least squares multiple regression versus LISREL. AB - The data of a specified path model using the variables of voice, perceived organizational support, being heard, and procedural justice were subjected to the two separate structural equation modeling analytic techniques--that of ordinary least squares regression and LISREL. A comparison of the results and differences between the analyses is discussed, with the LISREL approach being stronger from both theoretical and statistical perspectives. PMID- 11403344 TI - Organizational commitment, job satisfaction, and effort in the service environment. AB - Investigations of the causal relationship between organizational commitment and job satisfaction have yielded contradictory findings. Little empirical research has looked at this complex relationship in the context of work effort. The purpose of this study was to determine how these variables interact in the service environment. Using a sample of 425 employees in two service organizations, the author tested two structural equation models. The hypothesized model with organizational commitment as a moderator between job satisfaction and service effort fit better than a model with job satisfaction as moderator did. Conceptual implications are discussed, and suggestions for future research are made. PMID- 11403345 TI - Motive profiles of modern and traditional U.S. presidents. PMID- 11403346 TI - Vasodilatation, not hypotension, improves resistance vessel design during treatment of essential hypertension: a literature survey. AB - Correction of structural abnormalities in resistance arteries of patients with essential hypertension is a potential treatment goal, in addition to blood pressure reduction. However, available evidence from human as well as from animal studies indicates that antihypertensive therapy is not always accompanied by normalization of resistance vessel structure, despite normalization of blood pressure. Thus, blood pressure is not the only factor determining resistance vessel structure, and experimental studies show that several factors could play a role, including shear stress and hormonal stimulation. To date, there has been no systematic review of the many published papers which have studied the structural effects of antihypertensive therapy, and it is not known which conditions are best able to normalize resistance vessel structure. We have therefore made a survey of the available literature. The survey shows that change in blood pressure in indeed a poor indicator of change in resistance vessel structure. However, it is a remarkably consistent finding that normalization of resistance vessel structure is obtained with therapeutic regimens which reduce blood pressure by vasodilation rather than by lowering cardiac output Thus, to the extent that normalization of resistance vessel structure is deemed a goal of antihypertensive treatment, the survey points towards the importance of considering not only the treatment effect on blood pressure, but also the haemodynamic effects within patients with essential hypertension. PMID- 11403347 TI - The impact of genetic and environmental influences on blood pressure variance across age-groups. AB - OBJECTIVE: The purpose of the present investigation was to examine the age dependency of blood pressure heritability by use of the twin method in different age groups. DESIGN: In 272 (150 monozygous and 122 dizygous) twin pairs, aged 18 to 76 years, both conventional and ambulatory blood pressure were measured. After correction for possible confounders, model fitting was used to estimate heritability and 95% confidence limits in three age groups of similar size, i.e. 18-29, 30-39 and > or = 40 years. RESULTS: Heritability estimates were significant in each age group, ranging from 35 to 67% for the various blood pressure measurements. The estimates tended to decrease with increasing age, except for conventional diastolic blood pressure, but the intergroup differences did not reach statistical significance, despite adequate power. CONCLUSIONS: In conclusion, blood pressure heritability is significant in various age groups, but does not substantially change with advancing age. Twin analysis does not indicate that models for blood pressure regulation in various age groups should take into account the possibility of age-related changes in the expression of relevant genes, in the presence of relevant environmental agents, or in the susceptibility to the latter (gene-environment interaction). PMID- 11403348 TI - Independent predictors of isolated clinic ('white-coat') hypertension. AB - BACKGROUND: Hypertension guidelines recommend 24 h ambulatory blood pressure (ABP) monitoring in hypertensive subjects with suspected isolated clinic hypertension (ICH). However, the pre-test probability of ICH based on the distribution of its independent predictors has not yet been estimated in hypertensive subjects with mildly elevated blood pressure. OBJECTIVE: To ascertain the independent predictors of ICH in mildly hypertensive subjects. METHODS: In the setting of the HARVEST-PIUMA collaboration, we studied 1564 subjects with hypertension stage I. At entry, all subjects were untreated and all underwent ABP monitoring and echocardiography. Diabetes, hypertension grade > I, renal failure or previous cardiovascular morbid events were exclusion criteria. Clinic BP was 143/92 mmHg (SD 9/5) and 24 h ABP was 128/81 mmHg (SD 10/8). RESULTS: Prevalence of ICH (daytime ABP < 130 mmHg systolic and 80 mmHg diastolic) was 10.4%. In a multivariate logistic regression analysis, sex (P = 0.002), smoking (P = 0.038) and clinic diastolic BP (P = 0.0002) were the sole independent predictors of ICH according to the following equation: Y = 2.6438 + 0.5128 x sex (0 = men; 1 = women) + 0.4543 x current smoking (0 = yes; 1 = no) - 0.0531 x clinic diastolic BP (mmHg) and P (probability of ICH) = exp(Y)/[1 + (exp(Y)]. Left ventricular (LV) mass at echocardiography was a further independent predictor (P = 0.002) of ICH according to the following equation: Y= 3.4343 + 0.4603 x sex + 0.5989 x current smoking - 0.0482 x clinic diastolic BP - 0.0312 x LV mass [g/height (m)2.7]. LV mass was greater (P < 0.01) in the group with ambulatory hypertension [42.3 g/height (m)2.7] than in that with ICH [39.2 g/height (m)2.7] and not dissimilar between the ICH group and a control group of 370 healthy normotensive subjects [38.1 g/height (m)2.7]. CONCLUSIONS: In untreated subjects with stage I hypertension, ICH is most frequent among women, nonsmokers and subjects with low clinic BP and smaller LV mass. These findings allow identification of subjects with indication to ABP monitoring because of suspected ICH. PMID- 11403349 TI - Transient but not sustained blood pressure increments by occupational noise. An ambulatory blood pressure measurement study. AB - OBJECTIVE: Studies on the effects of chronic exposure to industrial noise on clinic blood pressure (BP) at rest have yielded inconsistent results. The aim of this study was to evaluate the effect of occupational noise exposure on ambulatory blood pressure (ABP) in normotensive subjects. METHODS: We studied 476 normotensive workers, aged 20-50 years (systolic blood pressure (SBP) < 140, diastolic blood pressure (DBP) < 90), at a metallurgical factory; 238 were exposed to high levels of noise (> 85 dB), while 238 were not exposed (< 80 dB). Clinical evaluation included measurements of casual BP (by standard mercury sphygmomanometer, Korotkoff sound phase I and V) and heart rate (HR) (by pulse palpation), body height and weight. All subjects underwent a 24 h non-invasive ABP monitoring (by SpaceLabs 90207 recorder; SpaceLabs, Redmond, Washington, USA) twice within 14 days: one during a normal working day and one during a non working day. Measurements were performed every 15 min. Computed analysis of individual recordings provided average SBP, DBP and HR values for 24 h, daytime working hours (0800-1700 h), daytime non-working hours (1700-2300 h) and night time (2300-0800 h). RESULTS: No significant difference in clinic SBP, DBP and HR was observed between exposed and non-exposed subjects. Results obtained by ABP monitoring showed in the exposed workers: (a) a higher SBP (by a mean of 6 mmHg, P < 0.0001 versus controls) and DBP (by a mean of 3 mmHg, P < 0.0001) during the time of exposure and the following 2 or 3 h, whereas no difference between the two groups was found during the non-working day; (b) an increase in HR, which was present not only during the time of exposure to noise (+3.7 beats-per-minute (bpm), P < 0.0001 versus controls), but also during the non-working hours (+2.8 bpm, P < 0.001) and during the day-time hours of the non-working day (+2.8 bpm, P < 0.003); (c) a significant increase in BP variability throughout the working day. CONCLUSIONS: These findings suggest that in normotensive subjects below the age of 50 years, chronic exposure to occupational noise is associated with a transient increase in BP, which is not reflected in a sustained BP elevation. The possible role of repeated BP and HR fluctuations due to frequent and prolonged exposure to noise in accounting for the higher prevalence of hypertension reported in noise-exposed workers above age 50 years, requires longitudinal studies to be clarified. PMID- 11403350 TI - Lessons from an unpleasant surprise: a biochemical strategy for the diagnosis of pheochromocytoma. AB - OBJECTIVE: To audit the performances of the analytes used in the diagnosis of pheochromocytoma and to present a graphical guideline to help the diagnosis. DESIGN: A 5 year retrospective study. SETTINGS: Laboratory and departments of a university hospital. PARTICIPANTS: In-patients, suspected of bearing a pheochromocytoma, were investigated for urinary metanephrines and catecholamines (photometric method) and vanillylmandelic acid, fractionated catecholamines and metanephrines [high pressure liquid chromatography (HPLC) coupled to electrochemical detection (ED)] urinary excretion. MAIN OUTCOME: Patients with a pheochromocytoma (24 out of 2003 patients) were diagnosed by the combination of normetanephrine and metanephrine determination. RESULTS: All analytes but dopamine were significantly elevated in patients with a pheochromocytoma. The area under the receiver operating characteristics (ROC) curves were the highest for total metanephrines, normetanephrine and metanephrine determinations. Because of analytical interferences in the metanephrines determination, the normetanephrine and metanephrine performed better. It is noteworthy that all pheochromocytomas had either normetanephrine or metanephrine levels above their respective optimal threshold (sensitivity 100%). The best optimal threshold performance was reached by the mean of three daily samples. Total or fractionated catecholamines or vanillylmandelic acid were less accurate tools. CONCLUSION: Amongst urinary tests, the combined use of HPLC/ED determination of normetanephrine and metanephrine seems the most effective screening strategy for the diagnosis of pheochromocytoma. The older total metanephrine photometric assay is grieved by analytical interferences. PMID- 11403351 TI - Non-invasive assessment of local arterial pulse pressure: comparison of applanation tonometry and echo-tracking. AB - OBJECTIVES: Pulse pressure is not constant throughout the arterial tree. Use of pulse pressure at one arterial site as surrogate for pulse pressure at another arterial site may be erroneous. The present study compares three non-invasive techniques to measure local pulse pressure: (i) internally calibrated readings from applanation tonometry, (ii) alternative calibration of pressure waves obtained with applanation tonometry and (iii) alternative calibration of arterial distension waves obtained with echo-tracking. Alternative calibration assumes mean and diastolic blood pressure constant throughout the large artery tree. DESIGN AND METHODS: Study 1 used invasive measurements in the ascending aorta as a reference method and internally calibrated tonometer readings and alternatively calibrated pressure waves at the common carotid artery as test methods. Study 2 used alternatively calibrated pressure waves as a reference method and alternatively calibrated distension waves and internally calibrated applanation tonometer readings as test methods. RESULTS: In study 1, pulse pressure from internally calibrated tonometer readings was 10.2+/-14.3 mmHg lower and pulse pressure from alternatively calibrated pressure waves was 1.8+/-5.2 mmHg higher than invasive pulse pressure. Pulse pressure from calibrated distension waves was 3.4+/-6.9 mmHg lower than pulse pressure from alternatively calibrated pressure waves. According to British Hypertension Society criteria, pulse pressure from the internally calibrated tonometer achieved grade D and pulse pressure from alternatively calibrated pressure waves achieved grade A. Pulse pressure from calibrated distension waves achieved grade B when alternatively calibrated pressure waves were used as a reference method. CONCLUSIONS: Pulse pressure obtained from alternatively calibrated tonometer-derived pressure waves and echo tracking-derived distension waves demonstrates good accuracy. Accuracy of pulse pressure from internally calibrated applanation tonometer readings at the carotid artery is poor. PMID- 11403352 TI - Effects of exercise training on home blood pressure values in older adults: a randomized controlled trial. AB - OBJECTIVE: To determine the effects of exercise training on the blood pressure (BP) values of older adults, using home blood pressure measurement. DESIGN: Randomized controlled trial. PARTICIPANTS: A total of thirty-nine free-living older adults (including 19 men) aged from 60-81 years with home systolic blood pressure > 120 mmHg and without significant cardiopulmonary-musculoskeletal disease, were randomly allocated to either 25 weeks of exercise training (exercise group) or to a control program (control group). MAIN OUTCOME MEASUREMENTS: Change in the 2-week averages of home systolic and diastolic blood pressure values measured with a validated automatic device before, during and after the intervention period. RESULTS: Compared with the control group, the exercise group showed a significant decrease in values for home systolic blood pressure (maximum between-group difference = 7.7 mmHg, P = 0.003) and home diastolic blood pressure (4.2 mmHg, P = 0.001). These changes were observed for both genders. CONCLUSIONS: Exercise training was effective for older adults in lowering home blood pressure values. This is the first trial to demonstrate the usefulness of home blood pressure measurement in examining the effect of exercise training on blood pressure values. PMID- 11403353 TI - Blood pressure is linked to salt intake and modulated by the angiotensinogen gene in normotensive and hypertensive elderly subjects. AB - OBJECTIVES: To evaluate salt sensitivity in elderly subjects with different forms of hypertension and controls and to investigate any modulation by genotype DESIGN: Randomized, double-blinded, placebo-controlled latin-square SETTING: Tertiary referral hospital PARTICIPANTS: Community subjects (n = 46) aged > or = 60 years classified as isolated systolic hypertension [ISH; systolic blood pressure (SBP) > or = 160, diastolic blood pressure (DBP) < 90 mmHg, n = 19], diastolic +/- systolic hypertension (SDH; DBP > or = 90 mmHg, n = 10) and normotension (SBP < 160, DBP < 90 mmHg, n = 17). INTERVENTION: Four 14 day treatments, 50, 100, 200 and 300 mmol/day of sodium chloride supplementation interspersed with 14 day washout periods on a salt-restricted diet. MAIN OUTCOME MEASURES: The 24 h blood pressure, heart rate, weight, urinary sodium and creatinine clearance measured during baseline, treatment and washout periods and angiotensinogen (AGT) and angiotensin converting enzyme (ACE) genotypes. RESULTS: For the entire cohort, the mean +/- standard error (SE) of change from baseline in SBP for 50, 100, 200 and 300 mmol/day salt was 7.7+/-2.4, 12.1+/-2.4, 16.6+/ 3.0, 18.5+/-2.6 mmHg, respectively. For DBP, the respective changes were: -0.1+/ 1.5, 2.4+/-1.6, 3.0+/-1.5, 5.8+/-1.7 mmHg. The increase in SBP among ISH subjects was significantly higher than among subjects in the SDH and normotensive groups (P < 0.05). AGT genotype influenced the effect of salt dose on the change in DBP (P = 0.006) but not SBP (P = 0.7). CONCLUSIONS: In healthy, older subjects, a linear increase in BP occurred with increasing salt dose, it appeared most pronounced in ISH subjects and could be modulated by AGT genotype. PMID- 11403354 TI - Genetic isolation of a blood pressure quantitative trait locus on chromosome 2 in the spontaneously hypertensive rat. AB - OBJECTIVES: Total genome scans of genetically segregating populations derived from the spontaneously hypertensive rat (SHR) and other rat models of hypertension have suggested the presence of quantitative trait loci (QTL) regulating blood pressure and cardiac mass on multiple chromosomes, including chromosome 2. The objective of the current study was to directly test for the presence of a blood pressure QTL on rat chromosome 2. DESIGN: A new congenic strain was derived by replacing a segment of chromosome 2 in the SHR between D2Rat171 and D2Arb24 with the corresponding chromosome segment from the normotensive Brown Norway rat. Arterial pressures were directly monitored in conscious rats by radiotelemetry. RESULTS: We found that the SHR congenic strain (SHR-2) carrying a segment of chromosome 2 from the Brown Norway rat had significantly lower systolic and diastolic blood pressures than the SHR progenitor strain. The attenuation of hypertension in the SHR-2 congenic strain versus the SHR progenitor strain was accompanied by significant amelioration of cardiac hypertrophy. CONCLUSIONS: These findings demonstrate that gene(s) with major effects on blood pressure exist in the differential segment of chromosome 2 trapped within the new SHR.BN congenic strain. PMID- 11403355 TI - Mildly oxidized low-density lipoprotein acts synergistically with angiotensin II in inducing vascular smooth muscle cell proliferation. AB - OBJECTIVES: Considerable attention has been focused on both mildly oxidized low density lipoprotein (mox-LDL) and highly oxidized LDL (ox-LDL) as important risk factors for cardiovascular disease. Further, angiotensin II (Ang II) appears to play a crucial role in the development of hypertension and atherosclerosis. We assessed the effect of oxidatively modified LDL and its major oxidative components, i.e., hydrogen peroxide (H2O2), lysophosphatidylcholine (LPC), and 4 hydroxy-2-nonenal (HNE) and their interaction with Ang II on vascular smooth muscle cell (VSMC) DNA synthesis. METHODS: Growth-arrested rabbit VSMCs were incubated in serum-free medium with different concentrations of native LDL, mox LDL, ox-LDL, H2O2, LPC, or HNE with or without Ang II. DNA synthesis in VSMCs was measured by [3H]thymidine incorporation. RESULTS: Ang II stimulated DNA synthesis in a dose-dependent manner with a maximal effect at a concentration of 1 micromol/l (173%). Ang II (0.5 micromol/l) amplified the effect of native LDL at 500 ng/ml, ox-LDL at 100 ng/ml, and mox-LDL at 50 ng/ml on DNA synthesis (108 to 234%, 124 to 399%, 129 to 433%, respectively). H2O2 had a maximal effect at a concentration of 5 micromol/l (177%), LPC at 15 micromol/l (156%), and HNE at 0.5 micromol/l (137%). Low concentrations of H2O2 (1 micromol/l), LPC (5 micromol/l), or HNE (0.1 micromol/l) also acted synergisitically with Ang II (0.5 micromol/l) in inducing DNA synthesis to 308, 304, or 238%, respectively. Synergistic interactions of Ang II (0.5 micromol/l) with mox-LDL, ox-LDL (both 50 ng/ml), H2O2 (1 micromol/l), LPC (5 micromol/l), or HNE (0.1 micromol/l) on DNA synthesis were completely reversed by the combined use of probucol (10 micromol/l), a potent antioxidant and candesartan (0.1 micromol/l), an AT1 receptor antagonist. CONCLUSIONS: Our results suggest that mox-LDL, ox-LDL, and their major components H2O2, LPC, and HNE act synergistically with Ang II in inducing VSMC DNA synthesis. A combination of antioxidants with AT1 receptor blockade may be effective in the treatment of VSMC proliferative disorders associated with hypertension and atherosclerosis. PMID- 11403356 TI - Vascular expression of angiotensin type 2 receptor in the adult rat: influence of angiotensin II infusion. AB - OBJECTIVE: The aim of this study was to investigate the relative role of the angiotensin type 1 (AT1) and type 2 (AT2) receptors in mediating angiotensin II induced regulation of AT2 receptor in mesenteric artery. DESIGN: Sprague-Dawley rats were infused with either angiotensin II or vehicle for 14 days at a dose of 58.3 ng/min. Ang II-infused rats were allocated to receive either an AT1 antagonist, valsartan at a dose of 30 mg/kg per day or the AT2 receptor antagonist PD123319 at a dose of 830 ng/min. METHODS: Gene and protein expression of the AT2 receptor in the mesenteric vasculature was assessed by quantitative reverse transcriptase polymerase chain reaction, immunohistochemistry and by in vitro autoradiography with a specific radioligand, 1251-CGP 42112B. RESULTS: The AT2 receptor mRNA and protein were detected in the mesenteric artery from adult rats. Both nuclear emulsion and immunohistochemical staining showed expression of the AT2 receptor in the adventitial and medial layers. Compared to control rats, angiotensin II infusion was associated with a significant increase in the AT2 receptor expression. Valsartan treatment significantly reduced AT2 receptor gene expression, with no significant effect of PD123319 on this parameter. CONCLUSIONS: This study confirms that the presence of the AT2 receptor in mesenteric arteries in adult rats, shows an up-regulation of the AT2 receptor following angiotensin II infusion and suggests a role for the AT1 receptor in this regulation. In view of the recently demonstrated effects of the AT2 receptor, these findings may be relevant to the role of the AT2 receptor in the pathophysiology of vascular remodeling. PMID- 11403357 TI - Angiotensin converting enzyme inhibitor restrains inflammation-induced vascular injury in mice. AB - OBJECTIVES: Recent advances in molecular genetics made the mouse model important for studying the genetic basis of hypertension and vascular diseases such as the components of the renin-angiotensin system. This study was undertaken to investigate the role of angiotensin converting enzyme (ACE) in the mouse vascular injury model. DESIGN AND METHODS: Inflammation-induced vascular injury was created by placing a polyethylene cuff around the femoral artery of 12-14-week old male FVB/N mice. Cuffed arteries were harvested and applied to reverse transcriptase-polymerase chain reaction analysis and immunohistochemistry for ACE. Subsequently, the effects of an ACE inhibitor, perindopril (3 mg/kg per day), on neointimal thickening were examined 2 weeks after cuff placement The influence of a nitric oxide synthase inhibitor, NG-nitro-L-arginine methyl ester (L-NAME, 20 mg/kg per day) on the effects of perindopril was also examined. RESULTS: ACE mRNA expression increased in a time-dependent manner up to 2 weeks after cuff placement. Immunoreactive ACE was localized in the endothelium in the intact artery, while positive staining was observed in the medial and neointimal layer as well as in the periadventitial region of the cuffed artery. The intimamedia area ratio was significantly decreased by perindopril treatment (vehicle, 0.75+/-0.10; perindopril, 0.32+/-0.04; P< 0.05). The effect of perindopril was abrogated by coadministration of L-NAME whereas L-NAME alone did not affect the intima-media ratio (L-NAME, 0.66+/-0.11; perindopril + L-NAME, 0.72+/-0.09). CONCLUSIONS: This study provides evidence that ACE plays a role in cuff-induced neointimal thickening in mice. Nitric oxide may contribute, at least in part, to the inhibitory effects of perindopril. PMID- 11403358 TI - Leptin interacts with heart rate but not sympathetic nerve traffic in healthy male subjects. AB - OBJECTIVE: Administration of leptin to animals increases sympathetic nerve activity and heart rate. We therefore tested the hypothesis that plasma leptin is linked independently to muscle sympathetic nerve activity (MSNA) and heart rate in healthy humans. METHODS: We measured plasma leptin, plasma insulin, body mass index (BMI), percent body fat, waist: hip ratio, MSNA, heart rate and blood pressure in 88 healthy individuals (50 men and 38 women). RESULTS: In men, plasma leptin concentration correlated significantly with BMI (r = 0.75, P < 0.001), percent body fat (r = 0.70, P< 0.001), waist: hip ratio (r = 0.69, P < 0.001), insulin (r = 0.37, P = 0.009), and age (r = 0.38, P = 0.006). Only BMI and waist: hip ratio were linked independently to plasma leptin concentration (r = 0.78, P < 0.001). Plasma leptin concentrations also correlated with heart rate (r = 0.39, P = 0.006) and mean arterial pressure (MAP; r = 0.38, P = 0.007), but not with MSNA (r = 0.17, P = 0.24). After adjustment for BMI and waist: hip ratio, plasma leptin concentration correlated significantly only with heart rate (r = 0.29, P = 0.04), and not with MAP (r = 0.21, P = 0.14). Individuals were divided into high leptin and low-leptin subgroups on the basis of plasma leptin concentrations adjusted for BMI and waist: hip ratio. Those with high leptin concentrations had significantly faster heart rates than those with low leptin. MAP and MSNA were similar in both subgroups. No relationship between leptin and either heart rate or MSNA was evident in women. CONCLUSIONS: In normal men, heart rate, but not MSNA, is linked to plasma leptin concentration. This sex-specific relationship between heart rate and plasma leptin is independent of plasma insulin, BMI, waist:hip ratio and percentage body fat. PMID- 11403359 TI - Altered growth response to prostaglandin E2 and its receptor signaling in mesangial cells from stroke-prone spontaneously hypertensive rats. AB - OBJECTIVE: Prostaglandin (PG) E2, a major arachidonic acid metabolite in the kidney, acts on four receptor subtypes (EP1, EP2, EP3 and EP4). One of major causes of end-stage renal failure is hypertensive renal disease, in which enhanced renal PGE2 production has been shown. In this study, to explore the pathophysiological significance of EP subtypes in the kidney, we examined the role of EP subtypes on proliferation of mesangial cells (MCs) from stroke-prone spontaneously hypertensive rats (SHRSPs), which show faster growth than those from normotensive Wistar-Kyoto rats (WKYs). DESIGN AND METHODS: Using MCs from SHRSPs and WKYs, we investigated DNA synthesis and its upstream event, the phosphorylation of extracellular signal-regulated kinase (ERK), together with the gene expression of EP subtypes. RESULTS: Sulprostone, an EP1 agonist, dose dependently increased DNA synthesis and the phosphorylation of ERK in MCs from both strains. The EP4 agonist, 11-deoxy-PGE1, inhibited sulprostone-induced phosphorylation of ERK in WKY-MCs. In contrast, 11-deoxy-PGE1 failed to inhibit the ERK activity in SHRSP-MCs. Interestingly, cAMP production mediated by EP4 was markedly attenuated in SHRSP-MCs as compared with that in WKY-MCs, despite the overproduction of endogenous PGE2 in SHRSP-MCs. Similar gene expressions of EP1 and EP4 and only faint expression of EP3 were detected in MCs from both strains. CONCLUSIONS: These results indicate that the PGE2/EP4 system counteracts the PGE2/EP1 system at the level of the intracellular signaling pathway. The altered EP4 signaling may play a critical role in the exaggerated mesangial growth in SHRSPs. PMID- 11403360 TI - The role of haem oxygenase in renal vascular reactivity in normotensive and hypertensive rats. AB - OBJECTIVE: To evaluate the contribution of the haem oxygenase-carbon monoxide (CO) system to renal vascular tone in normotensive Wistar rats and in the stroke prone spontaneously hypertensive rats (SHR-SPs). METHODS: An isolated perfused rat kidney preparation was used in which perfusion pressure/phenylephrine dose vasoconstrictor responses were generated. Haemin was given 24 h previously, to induce haem oxygenase-1 (HO-1), and L-NAME (N-omega-nitro-L-arginine methyl ester) was given to block nitric oxide (NO) production. RESULTS: Haemin pretreatment attenuated the phenylephrine-induced rise in perfusion pressure (P < 0.05) but L-NAME had no effect on the magnitude of the renal vasoconstrictor responses to phenylephrine. This suggested that the effect of haemin incubation on renovascular responses did not involve NO production. Pretreatment of the rats with the haem oxygenase inhibitor, tin protoporphyrin IX (SnPP-IX) had no effect on either the basal tone or the phenylephrine-induced contractions in the renal vasculature. By contrast, the renovascular responses to phenylephrine in haemin treated animals were restored following the coadministration of SnPP-IX. Haemin administration in the SHR-SPs caused a significantly greater reduction in the renovascular responses to phenylephrine compared to those in the normotensive animals (P < 0.05). CONCLUSIONS: Following induction of HO-1, the HO-CO system plays an important role in the regulation of renal responses to an adrenergically induced vasoconstrictor challenge. Moreover, the renal vascular bed of hypertensive animals exhibited a greater propensity to upregulate the HO-CO system, which may provide an important counteractive role against the elevation of blood pressure in hypertension. PMID- 11403361 TI - Appropriate or inappropriate left ventricular mass in the presence or absence of prognostically adverse left ventricular hypertrophy. AB - OBJECTIVES: To evaluate whether assessment of appropriateness of left ventricular mass (LVM) adds to the traditional definition of left ventricular hypertrophy (LVH). DESIGN: Cross-sectional, relational. METHODS: Echocardiographic LVH and appropriateness of LVM were studied in 562 subjects (231 normotensive controls, aged 35+/-11 years, 142 women; 331 hypertensive patients, aged 47+/-11 years, 135 women) classified on the basis of either the presence or the absence of both LVH (LVM index > or = 51 g/m2.7) and inappropriate LVM (LVM > 128% of the value predicted by an equation including age, sex and stroke work). RESULTS: Body mass index was comparable in hypertensive patients and controls. Hypertensive patients without LVH but with inappropriate LVM (n = 21) had higher relative wall thickness and total peripheral resistance than all other groups, whereas cardiac output was lower (all P < 0.001). Midwall mechanics was normal with appropriate LVM, independently of presence of LVH, whereas it was depressed in inappropriate LVM, either with or without LVH (both P < 0.0001). There was no substantial difference in ejection fraction among controls and hypertensive groups. Stress corrected midwall shortening was more closely related to deviation of LVM from the value appropriate for stroke work, body size and gender (r = -0.56, P < 0.0001) than to LVM index (r = -0.26). CONCLUSIONS: Inappropriate LVM is associated with concentric geometry, high peripheral resistance and depressed wall mechanics. The deviation of LVM from the value appropriate for stroke work, body size and sex correlates with measures of myocardial function better than LVM. PMID- 11403362 TI - Postischemic apoptosis and functional recovery after angiotensin II type 1 receptor blockade in isolated working rat hearts. AB - OBJECTIVE: To determine whether chronic angiotensin (AngII) type I receptor (AT1R) blockade inhibits cardiomyocyte (CM) apoptosis and attenuates left ventricular (LV) dysfunction after ischemia-reperfusion (IR) in the isolated working rat heart. METHODS: Postischemic recovery of LV developed pressure, the apoptotic index (terminal deoxynucleotidyl transferase (TdT)-mediated dUTP in situ nick end labeling or TUNEL assay), and changes in expression of apoptotic markers Bcl-2, Bax, p53 and caspase-3 (Western immunoblots) were measured after IR (50 min aerobic perfusion; 25 min global ischemia; 40 min reperfusion) in working rat hearts that were randomized to five groups of six each along 1 week or 3 week pretreatment arms: sham (no drug, no perfusion); no drug, aerobic perfusion; and oral AT1R blockers losartan (30 mg/kg per day) or UP269-6 (3 mg/kg per day), or no drug before IR. RESULTS: Compared to the no drug group after IR, losartan (not UP269-6) preserved functional recovery in 1 and 3 week groups. However, both losartan and UP269-6 reduced the apoptotic index and normalized the increase in Bax, decrease in Bcl-2 and increase in p53 and caspase-3 after IR. A bell-shaped relation between apoptosis and functional recovery after IR was flattened by AT1R blockade. CONCLUSION: The results indicate that IR is associated with LV dysfunction and CM apoptosis involving activation of p53, caspase-3, and increased Bax/Bcl-2 ratio in the working rat heart. Importantly, chronic AT1R blockade inhibited the apoptosis and changes in expression of the markers without improving functional recovery, implying that decrease in apoptosis does not necessarily translate into decreased LV dysfunction. PMID- 11403363 TI - Base heart rate during sleep in hypertensive and normotensive subjects. AB - OBJECTIVE: Many studies have reported that the resting heart rate (HR) is intimately related to the prognosis of cardiovascular disease. However, HR in the waking state is influenced by psychological and physical activity and has low reproducibility. To measure the base HR (minimum and stable HR during sleep) with ease, we developed a wristwatch-type HR recorder. We then investigated the pathophysiological significance of the base HR. SUBJECTS AND METHODS: The subjects were 158 patients (82 men and 76 women) with essential hypertension (HT) on no medication and 204 normotensive (NT) subjects (105 men and 99 women, matched for age). On the basis of pulse waves from the wristwatch-type photoelectric plethysmograph, using a blue sensor with a wavelength of 450 nm, HR was recorded easily minute by minute during sleep time. In addition, 40 (23 men and 17 women) of the HT patients were hospitalized. Their cardiac index (CI) and stroke volume index (SVI) were measured by means of the cuvette method during waking time and night-time deep sleep when their HR had reached a minimum level. RESULTS: Mean base HR (HRo) in NT subjects was 49+/-4 beats per minute (bpm) and tended to rise with increasing age (r = 0.51, P < 0.01). In HT patients, HR0 was significantly higher: 60+/-7 bpm (P < 0.01). CI of HT patients, measured during night-time deep sleep, maintained a stable value of 2.45+/-0.36 l/min per m2. SVI in patients whose HR0 was above 60 was significantly lower than that in patients whose HR0 was below 60 (37+/-4.5: 45+/-4.8 ml/m2, P < 0.01). Corresponding differences in mean HR and SVI during waking or night-time, became smaller. CONCLUSIONS: These findings suggest that there is a close relation between base HR and cardiac function caused by such conditions as age and hypertension. This relation became indistinct during waking time. Moreover, the wristwatch-type HR recorder appears to be a useful tool for measuring base HR. PMID- 11403364 TI - Rationale, design, methods and baseline demography of participants of the Anglo Scandinavian Cardiac Outcomes Trial. ASCOT investigators. AB - OBJECTIVE: To test the primary hypothesis that a newer antihypertensive treatment regimen (calcium channel blocker +/- an angiotensin converting enzyme inhibitor) is more effective than an older regimen (beta-blocker +/- a diuretic) in the primary prevention of coronary heart disease (CHD). To test a second primary hypothesis that a statin compared with placebo will further protect against CHD endpoints in hypertensive subjects with a total cholesterol < or = 6.5 mmol/l. DESIGN: Prospective, randomized, open, blinded endpoint trial with a double blinded 2 x 2 factorial component. SETTING: Patients were recruited mainly from general practices. PATIENTS: Men and women aged 40-79 were eligible if their blood pressure was > or = 160 mmHg systolic or > or = 100 mmHg diastolic (untreated) or > or = 140 mmHg systolic or > or = 90 mmHg diastolic (treated) at randomization. INTERVENTIONS: Patients received either amlodipine (5/ 10 mg) +/- perindopril (4/8 mg) or atenolol (50/ 100 mg) +/- bendroflumethiazide (1.25/2.5 mg) +K+ with further therapy as required to reach a blood pressure of < or = 140 mmHg systolic and 90 mmHg diastolic. Patients with a total cholesterol of < or = 6.5 mmol/l were further randomized to receive either atorvastatin 10 mg or placebo daily. MAIN OUTCOME MEASURE: Non-fatal myocardial infarction (MI) and fatal coronary heart disease (CHD). RESULTS: 19 342 men and women were initially randomized, of these 10297 were also randomized into the lipid-lowering limb. All patients had three or more additional cardiovascular risk factors. CONCLUSIONS: The study has 80% power (at the 5% level) to detect a relative difference of 20% in CHD endpoints between the calcium channel blocker-based regimen and the beta blocker-based regimen. The lipid-lowering limb of the study has 90% power at the 1% level to detect a relative difference of 30% in CHD endpoints between groups. PMID- 11403365 TI - Effects of individual risk factors on the incidence of cardiovascular events in the treated hypertensive patients of the Hypertension Optimal Treatment Study. HOT Study Group. AB - BACKGROUND: The Hypertension Optimal Treatment (HOT) Study has provided information about cardiovascular events in 18790 hypertensives, subjected to pronounced blood pressure (BP) lowering for a mean of 3.8 years. The HOT study data have subsequently been analysed after stratification of the patients according to global cardiovascular risk, and it has been found that, despite intensive blood pressure lowering in all risk strata, morbid event rates increased with increasing risk stratum. OBJECTIVES: Previously analysed global risk strata were based on combinations of risk factors. The analyses presented here were intended to provide information on the relative role that the presence of each individual factor may have in increasing cardiovascular risk, despite good BP control. METHODS: Risk ratios (RR) for patients with and those without a risk factor were calculated with 95% confidence intervals (CI) using a Cox proportional hazard model, and adjusted for all variables except the one under examination. RESULTS: For all risk factors considered and for all types of event, RR were always greater than 1, indicating a greater risk in the presence, compared with that in the absence of each factor. The male gender was a statistically significant risk for cardiovascular (CV) events, CV and total mortality and particularly for myocardial infarction (MI); age > or = 65 years for CV events, stroke, CV and particularly total mortality; smoking for all events analysed, but particularly for total mortality (twice higher in smokers than in non-smokers); high serum cholesterol (> 6.8 mmol/l) for CV events, MI and CV mortality; high serum creatinine (> 155 micromol/l) for CV events, stroke, CV and total mortality; diabetes for CV events, stroke, total mortality and particularly CV mortality; and ischaemic heart disease for all events analysed. Adjusted RR were often close to or greater than 2. CONCLUSIONS: Each of the risk factors considered was found to be an important cause of residual risk, despite good BP control. These findings emphasize the importance of addressing other correctable risk factors, e.g. smoking, hypercholesterolaemia and diabetes, as well as rigorous control of blood pressure, and of initiating antihypertensive therapy before cardiovascular and renal damage becomes manifest. PMID- 11403366 TI - Efficacy of an angiotensin II receptor antagonist in managing hyperaldosteronism. AB - OBJECTIVE: To determine whether an angiotensin II receptor antagonist decreases blood pressure in patients with hyperaldosteronism and hypertension who are taking other antihypertensive agents. DESIGN: A double-blind randomized placebo controlled crossover study. PATIENTS AND METHODS: Blood pressure and hormonal responses to 2-week courses of placebo/irbesartan (150 mg/day given by mouth at 08.05 h) were assessed in 10 patients with hyperaldosteronism. Clinic blood pressure was measured by sphygmomanometer, and plasma concentrations of aldosterone, cortisol, angiotensin II, electrolytes and renin activity (PRA) were determined weekly. Automated 24 h ambulatory blood pressure recordings were made at the end of the active and placebo phases. RESULTS: Irbesartan caused a post dose decrease in ambulatory blood pressure (systolic, P = 0.02; diastolic, P = 0.05) in the period from 10.00 h to 20.00 h. Clinic blood pressure, measured at trough, was not significantly decreased. Plasma aldosterone decreased (P < 0.03) and PRA increased (P < 0.04) in the first week of active treatment with irbesartan, but differences between the placebo and active-treatment groups were not significant in the second week. There were no significant changes in plasma concentrations of angiotensin II, cortisol or potassium in either week. In the second week of irbesartan treatment, there were associations between change in plasma aldosterone and maximal change in ambulatory blood pressure (systolic and diastolic). CONCLUSION: Irbesartan has a role in combination antihypertensive treatment of patients with hyperaldosteronism. PMID- 11403367 TI - Regression of left ventricular hypertrophy in human hypertension with irbesartan. AB - BACKGROUND: The Swedish irbesartan left ventricular hypertrophy investigation versus atenolol (SILVHIA). OBJECTIVE: Angiotensin II induces myocardial hypertrophy. We hypothesized that blockade of angiotensin II subtype 1 (AT1) receptors by the AT1-receptor antagonist irbesartan would reduce left ventricular mass (as measured by echocardiography) more than conventional treatment with a beta blocker. DESIGN AND METHODS: This double-blind study randomized 115 hypertensive men and women with left ventricular hypertrophy to receive either irbesartan 150 mg q.d. or atenolol 50 mg q.d. for 48 weeks. If diastolic blood pressure remained above 90 mmHg, doses were doubled, and additional medications (hydrochlorothiazide and felodipine) were prescribed as needed. Echocardiography was performed at weeks 0, 12, 24 and 48. RESULTS: Baseline mean blood pressure was 162/ 104 mmHg, and mean left ventricular mass index was 157 g/m2 for men and 133 g/m2 for women. Systolic and diastolic blood pressure reductions were similar in both treatment groups. Both irbesartan (P < 0.001) and atenolol (P< 0.001) progressively reduced left ventricular mass index, e.g. by 26 and 14 g/m2 (16 and 9%), respectively, at week 48, with a greater reduction in the irbesartan group (P = 0.024). The proportion of patients who attained a normalized left ventricular mass (i.e. < or = 131 g/m2 for men and < or = 100 g/m2 for women) tended to be greater with irbesartan (47 versus 32%, P = 0.108). CONCLUSIONS: Left ventricular mass was reduced more in the irbesartan group than in the atenolol group. These results suggest that blocking the action of angiotensin II at AT1-receptors may be an important mechanism, beyond that of lowering blood pressure, in the regulation of left ventricular mass and geometry in patients with hypertension. PMID- 11403368 TI - Defining ambulatory and home blood pressure normality: further considerations based on data from the PAMELA study. PMID- 11403369 TI - Are reliability, reproducibility and validity the correct terms to assess the correctness of dietary studies? AB - Nutritional studies often use the terms reliability, reproducibility and validity to indicate the correctness of the study. These terms do not appear to have a universal meaning to all researchers. The components of a dietary study are the input, the data collection instrument and the compiled data. Frequently the data collection questionnaire/tool/instrument is tested for reliability, reproducibility or validity. The data collection questionnaire/tool/instrument is simply a structure, a vehicle for gathering data. An argument is presented that demonstrates the reasons that such a structure cannot be tested for reliability, reproducibility or validity. The logical approach to the use of the terms reliability, reproducibility and validity is presented. Reliability refers to the input component of the study, reproducibility may or may not lead to strengthening the study and validity refers to the truthfulness of the database generated. Validity must be derived from reliable and reproducible data. PMID- 11403370 TI - Assessment of nutritional status using anthropometric methods on 1-4 year old children in an urban ghetto in Lagos, Nigeria. AB - This study assessed the nutritional status using anthropometry of 1-4 year old children in an urban slum in the Mushin Local Government Area of Lagos State, with a view to determining the impact of urbanization on child health. A total of 365 children were enrolled using multistage random sampling techniques. Anthropometric measurements used were weight and height. Height-for-age, weight for-height, and weight-for-age Z-scores below -2.00 SD of the reference NCHS standard were used to define stunting, wasting and underweight, respectively. The study revealed a prevalence of underweight of 39.2%, stunting of 34.5% and wasting of 21.9%. The mean of weight-for-age, height-for-age, weight-for-height and mid-upper-arm-circumference were less than the mean of the NCHS reference population. This difference might be due to the socio economic backgrounds of the two populations. Using the modified Wellcome Classification of malnutrition, 37.8% of the children were malnourished. Most subjects came from homes with inadequate water supply and poor refuse disposal methods. PMID- 11403371 TI - Reducing waiting lists for hospital admission: community nutrition services reduce the need for hospital beds. AB - The number of hospital beds needed is greatly increased by the malnutrition of patients before admission and after discharge. Malnourished patients spend longer in hospital and are more likely to die following surgery or other treatment. Extensive studies show that low blood serum albumin, indicating protein deficiency, is a major risk factor for morbidity and mortality among hospital patients. Community food and nutrition services are extending throughout the developed world as part of public health policies. Such services can reduce costs by reducing the need for hospital beds, can reduce waiting lists and save the lives of many hospital patients. Preventive nutrition services can give financial benefits much exceeding costs. PMID- 11403372 TI - Sociocultural practices influencing under five nutritional status in an urban community in Osun State, Nigeria. AB - Three hundred and forty four preschool children from Ilare district of Ife Central Local Government Area of Osun State, Nigeria were assessed for nutritional status using selected and sensitive anthropometric techniques. The sociocultural practices of the parents were also examined. Approximately 53.8% and 54.7% of the children were identified to be malnourished and stunted respectively. The socioeconomic situation of the family was identified to have a significant influence on the child's nutritional status. Female children were observed not to be as adequately nourished as their male counterparts. However parity and food taboo had no significant influence on the child's nutritional status. The need for improvement in the socioeconomic status of the family and education of the community on the importance of female nutrition are discussed. PMID- 11403373 TI - Modification and improvement of nutritive quality of cornpap "Ogi" with cowpea and groundnut milk. AB - The study aimed at improving the nutritive quality of cornpap, "Ogi", the most popular infant feed in Nigeria. Milk samples of cowpea, groundnut and soyabean and their complementations with cornpap were assessed chemically, organoleptically and anthropometrically. The protein contents of milk from cowpea, groundnut and soyabean were 1.18, 1.6, and 1.23% respectively. On separate complementation of the milk sources with cornpap, the low protein content of "ogi" was increased from 0.3% to 2.79% with cowpea; 3.0% with groundnut and 3.64% with soyabean. The fat content of the complementary feeds also increased remarkably. The anthropometric study revealed that there were significant differences P = 0.001 between the mean weight of children aged 13 to 18 (92 +/- 1.2) and 19 to 24 months (9.9 +/- 1.3) who were fed the three sources of milk with cornpap, and the control group of the same age groups (8.5 + 1.5; 9.3 + 1.11) who received nutrition education and no complementary feeds. Of the three sources of milk, groundnut milk was ranked as the most likeable, the easiest to prepare and the least costly by the nursing mothers. This study has shown that some of the deep rooted cultural food taboos that prohibit locally available and nutritious feeds to infants could be changed through practical food demonstrations. PMID- 11403374 TI - Promotion of exclusive breastfeeding (EBF): the need to focus on the adolescents. AB - This study was designed to assess the knowledge and attitude towards exclusive breast feeding among 377 female students of School of Health Technology, Ilesha and to compare their responses with 60 primigravidae attending antenatal clinic in Ile-Ife, Nigeria. The ages of the subjects ranged from 15 to 34 years. Data were collected using pretested structured questionnaire. Approximately 47% of the total population were grouped under low level of knowledge of exclusive breastfeeding. There was no significant relationship in terms of knowledge between the two groups. There was, however, a significant relationship between the age of subjects and increased level of knowledge about EBF. Seventy percent of the primigravidae were graded as having poor attitudes as compared with 18% of the female students. About 42% of the total population would give water and glucose D water to neonates within 72 hours after delivery. These findings further suggest that planners of the Baby Friendly Initiative need to focus more on adolescents and the primigravidae in the promotion of breastfeeding. PMID- 11403375 TI - Waters-edge evolution. PMID- 11403376 TI - Food consumption pattern and nutritional problems in the Islamic Republic of Iran. PMID- 11403377 TI - Do Blastocystis hominis colony forms undergo programmed cell death? AB - Ultrastructural observations were made on colony forms of the protozoan parasite, Blastocystis hominis. Cross-sections of entire colonies were observed by TEM. Cells within a colony were heterogeneous in morphology, consisting of vacuolar, amoeboid, multivacuolar and unusual forms. Dying cells appeared to be in the process of fragmenting into numerous membranebound vesicles, giving rise to empty spaces within the colony. Interestingly, older colonies appeared to show cell fragmentation which resulted in larger, membranebound structures. Numerous cytoplasmic inclusions were present in the central vacuole of some cells, with many containing mitochondria. Amoeboid forms were observed to harbour small membrane-bound vesicles in endosome-like compartments. Other unusual features included margination of chromatin material and distinct blebbing of nuclei. These ultrastructural features suggest that B. hominis colony forms perhaps undergo a form of programmed cell death. PMID- 11403378 TI - Cryptosporidium parvum genotype 2 infections in free-ranging mountain gorillas (Gorilla gorilla beringei) of the Bwindi Impenetrable National Park, Uganda. AB - For behavioral research and due to growing ecotourism, some populations of free ranging mountain gorillas (Gorilla gorilla beringei) have become habituated to humans. Molecular analysis of two Cryptosporidium sp. oocyst isolates originating from two human-habituated gorilla groups and two oocyst isolates from non habituated gorillas yielded positive identification of C. parvum Genotype 2 (G2; i.e., "cattle", "animal-adapted", or "zoonotic"). As G2 is cross-transmissible between humans and animals, C. parvum infections can be propagated in the habitats of human-habituated, free-ranging gorillas through both zoonotic and anthroponotic transmission cycles. PMID- 11403379 TI - Inhibition of encystation of Entamoeba invadens by wortmannin. AB - Using an axenic encystation system in vitro, we examined the effect of wortmannin, a potent inhibitor of phosphatidylinositol 3-kinase (PI 3-kinase), which is a signaling molecule responsible for numerous cellular responses, on the encystation of Entamoeba invadens. Wortmannin inhibited both encystation and growth of E. invadens strain IP-1 in a dose-dependent manner, the former being more resistant to the drug than the latter. There was little decrease in the number of trophozoites after 3 days of culture in encystation medium containing wortmannin; and the cells remained motile, suggesting that the inhibitory effect of the drug on encystation was not due to its toxic effect on trophozoites. The addition of wortmannin after the induction of encystation was also inhibitory for encystation. Trophozoites incubated for 1 day in encystation medium with wortmannin did not encyst after removal of the drug, suggesting that the drug effect was not reversible in encystation medium. In contrast, trophozoites cultured in growth medium with wortmannin did encyst after their transfer to encystation medium without the drug. Encystation with wortmannin was more strongly inhibited among trophozoites grown in the presence of the drug than among those grown in the absence of the drug. The process of cyst maturation was slightly affected by wortmannin. These results suggest a possible role for PI 3 kinase in the signaling involved in the encystation of E. invadens. PMID- 11403380 TI - The role of saliva of Anopheles stephensi in inflammatory response: identification of a high molecular weight neutrophil chemotactic factor. AB - Mosquito bites can elicit dermal hypersensitivity reactions, but little is known about the chemotactic factors for host leukocytes in mosquito saliva. In this study, we determined that saliva from a malarial vector mosquito, Anopheles stephensi, possesses intense neutrophil chemotactic activity. In contrast, the midgut extract had only marginal neutrophil chemotactic activity. Eosinophil chemotactic activity was detected in the midgut but not in the saliva. According to the results of size-exclusion HPLC on a G3000SW column and Western blot analysis, the apparent molecular weight (MW) of the main neutrophil chemotactic factor (NCF) was estimated to be 200 kDa. NCF could bind with IgG from the pooled serum of Solomon islanders, whereas not with that of healthy Japanese. NCF activity was increased upon heating to 56 degrees C for 30 min or protease digestion, whereas it was affected by periodate treatment. Protease-digested NCF and naive NCF bound to lentil lectin-Sepharose, and both were eluted with a competitive sugar, methyl-alpha-D-glucoside. These results indicate that A. stephensi saliva-derived NCF is a high MW glycoprotein, and its protein moiety is important for neutrophil chemotactic activity. This NCF is thought to contribute to the inflammatory reactions through the accumulation of neutrophils at the site of the mosquito bite. PMID- 11403381 TI - Recombinant dissection of myosin heavy chain of Toxocara canis shows strong clustering of antigenic regions. AB - Myosins from nematode parasites elicit strong humoral and cellular immune responses and have been investigated as vaccine candidates. In this study we cloned and sequenced a cDNA coding for myosin heavy chain from Toxocara canis, a nematode parasite of canids which may also infect humans and cause various unspecific symptoms. To determine the major antigenic regions the myosin heavy chain was systematically dissected into ten overlapping recombinant fusion polypeptides which were purified by metal chelate chromatography. Single fragments were then tested for their IgG reactivity in sera from toxocarosis patients and healthy probands. Two regions, one region at the mid to carboxy terminal end of the head domain and one region in the rod domain, were identified as major antigens, which in combination were positive with 86% of the sera. The other domains were less reactive. This shows that the patients' IgG reactivity was not directed evenly against all parts of the molecule, but was rather clustered in few regions. PMID- 11403382 TI - Heterologous expression and functional characterization of thioredoxin from Fasciola hepatica. AB - The full thioredoxin coding sequence from Fasciola hepatica has been cloned into the pGEX-2T expression vector and produced in Escherichia coli as a fusion protein. The recombinant protein proved to be biologically active, using an insulin reduction assay, and was also able to activate thioredoxin peroxidase from F. hepatica. These observations suggest that this protein could participate in a redox cascade involved in the maintenance of cell homeostasis as well as in parasite protection against reactive oxygen species produced by the host. PMID- 11403383 TI - Cellular immune responses in mice immunized with Anisakis simplex larval antigens. AB - Cellular immune responses to Anisakis simplex L3 antigens were investigated in BALB/c mice injected subcutaneously with a homologous crude extract (CE). Popliteal lymph nodes (PLN) were found to be increased in size and weight after A. simplex CE footpad injection. The effects of A. simplex CE in vitro proliferation were assayed with non-fractionated PLN cells or nylon-wool purified T cells derived from pooled lymph node cells of mice subcutaneously injected with CE. Spleen cells from immunized animals (antigen alone, or larva alone, or antigen plus larva) were studied by flow cytometry. The immunization induced a high proportion of CD4 + and TCRalphabeta + T cells. The number of B cells (CD45 + and TCRalphabeta-) in pre-immunized and infected mice was lower than that observed in animals subjected to infection only. The number of CD4+ T cells increased in the infected and in the pre-immunized and infected mice. In the latter, a decrease of CD8a + T cells was noted. The greatest increase in CD8a+ and TCRalphabeta- T cells was found in mice that had been subjected to infection only. Histological analysis showed that the most prominent lesions were gastric and intestinal in animals infected orally with one larva. PMID- 11403384 TI - Involvement of CD 8+ and CD 3+ lymphocytes in the transport of Eimeria necatrix sporozoites within the intestinal mucosa of chickens. AB - The phenotype of cells transporting sporozoites of Eimeria necatrix during a primary infection was determined using a panel of six monoclonal antibodies to various chicken lymphocyte surface markers. Sporozoites and cells harboring them were examined at 8, 12 and 18 h postinfection using two-color immunoflorescence and confocal microscopy. The majority of parasites observed within lymphocytes were found in CD 8 + (15%) or CD 3+ (13-22%) cells at all time periods examined. Smaller numbers were found within deltagamma TCR+ (5%) and alphabeta TCR+ (5%) lymphocytes. No sporozoites were found within CD 4+ or IgM+ lymphocytes at any of the time periods. PMID- 11403385 TI - Autofluorescence microscopy for the detection of nematode eggs and protozoa, in particular Isospora suis, in swine faeces. AB - Parasites from swine faeces were examined for autofluorescence. Oocysts of Eimeria polita, E. scabra and Isospora suis, cysts of Balantidium coli and eggs of Oesophagostomum dentatum, Strongyloides ransomi and Trichuris suis (but not those of Ascaris suum) emitted light after excitation with UV light. I. suis oocyst counts in McMaster chambers utilising autofluorescence were compared to those from conventional bright field microscopy. Similarly, faecal smears containing I. suis were examined using the same techniques. Autofluorescence was superior to bright field microscopy in detecting oocysts after flotation and was highly significantly more sensitive when direct smears were examined. PMID- 11403386 TI - Taking more care in using different species concepts--an opinion. PMID- 11403387 TI - Molecular identification of Wolbachia from the filarial nematode Mansonella ozzardi. AB - Mansonella ozzardi, a filarial parasite of humans in Latin America, has been shown to harbour intracellular bacteria not yet identified. Here we show that these bacteria, like those of other filarial nematodes, belong to the genus Wolbachia (alpha 2 Proteobacteria; Rickettsiales). Their unambiguous placement in the Wolbachia group was shown by 16S rDNA sequence analysis. However, the exact position of the Wolbachia from M. ozzardi relative to the other wolbachiae is not clear. Indeed, 16S rDNA sequence analysis places this bacterium at a deep branch in Wolbachia evolution. It is interesting that analysis of the 5S rDNA gene spacer of the nematode host also suggests that the genus Mansonella, together with the genus Loa, could represent a deep-branching lineage in filarial evolution. PMID- 11403388 TI - Synthesis of multiple N-acylhomoserine lactones is wide-spread among the members of the Burkholderia cepacia complex. AB - Seventy strains of the Burkholderia cepacia complex, which currently comprises six genomic species, were tested for their ability to produce N-acylhomoserine lactone (AHL) signal molecules. Using thin layer chromatography in conjunction with a range of AHL biosensors, we show that most strains primarily produce two AHLs, namely N-octanoylhomoserine lactone (C8-HSL) and N-hexanoylhomoserine lactone (C6-HSL). Furthermore, some strains belonging to B. vietnamiensis (genomovar V) produce additional long chain AHL molecules with acyl chains ranging from C10 to C14. For B. vietnamiensis R-921 the structure of the most abundant long chain AHL was confirmed as N-decanoylhomoserine lactone (C10-HSL) by liquid chromatography-mass spectrometry (LC-MS) in combination with total chemical synthesis. Interestingly, a number of strains, most notably all representatives of B. multivorans (genomovar II), did not produce AHLs at least under the growth conditions used in this study. All strains were also screened for the production of extracellular lipase, chitinase, protease, and siderophores. However, no correlation between the AHL production and the synthesis of these exoproducts was apparent. Southern blot analysis showed that all the B. cepacia complex strains investigated, including the AHL-negative strains, possess genes homologous to the C8-HSL synthase cepI and to cepR, which encodes the cognate receptor protein. The nucleotide sequence of the cepI and cepR genes from one representative strain from each of the six genomovars was determined. Furthermore, the cepI genes from the different genomovars were expressed in Escherichia coli and it is demonstrated that all genes encode functional proteins that direct the synthesis of C8-HSL and C6-HSL. Given that cepI from the B. multivorans strain encodes a functional AHL synthase, yet detectable levels of AHLs were not produced by the wild-type, this suggests that additional regulatory functions may be present in members of this genomovar that negatively affect expression of cepI. PMID- 11403389 TI - Subtraction hybridization in microplates: an improved method to generate strain specific PCR primers. AB - An improved subtraction hybridization technique was developed and evaluated. The hybridization is performed in a microplate with the subtractor-DNA immobilized in the plate while the probe-DNA is in solution. After hybridization the probe specific DNA can easily be removed from the microwell and submitted to further analysis. This new technique has been successfully applied to generate several strain-specific PCR-primers for Lactococcus lactis subsp. lactis, Pediococcus spec., Saccharomyces spec. and Listeria monocytogenes. PMID- 11403390 TI - Antimicrobial susceptibility of intestinal bacteria from Swiss poultry flocks before the ban of antimicrobial growth promoters. AB - From the crop and the caecum of Swiss broilers slaughtered between November 1997 and January 1998, Escherichia coli, enterococci, staphylococci, lactobacilli and Campylobacter species were isolated. After identification to the genus or species level, their minimal inhibitory concentrations (MIC's) for several clinically used antimicrobial agents were determined with the E-Test stripes and compared to those from studies in other European countries. All strains of Enterococcus faecalis (n = 38), E. faecium (27), staphylococci (n = 39) and lactobacilli (n = 14) showed a hundred percent resistance against bacitracin which was included in the feed of the mother animals, but not in the feed of the investigated animals. E.coli strains (n = 60) showed higher resistance incidences than in comparable studies from Finland and Denmark, but lower than those in studies from Italy and Germany. In staphylococci, low resistance rates were observed. A high susceptibility of the 13 Campylobacter jejuni strains was found against therapeutically used antimicrobials. These data can be used as a baseline to determine antibiotic resistance rates after implementation of the growth promotor ban in 1999 in Switzerland. PMID- 11403391 TI - Comparison of the antimicrobial tolerance of oxytetracycline-resistant heterotrophic bacteria isolated from hospital sewage and freshwater fishfarm water in Belgium. AB - The aim of this study was to investigate the relationship between antimicrobial tolerance and taxonomic diversity among the culturable oxytetracycline-resistant (Ot(r)) heterotrophic bacterial population in two Belgian aquatic sites receiving wastewater either from human medicine or from aquaculture. The study of Ot(r) heterotrophs and mesophilic Aeromonas spp. allowed comparison of tolerance data at the intergenus as well as at the intragenus level. In total, 354 independently obtained Ot(r) isolates were subjected to antimicrobial tolerance testing and identified by GLC analysis of their cellular fatty acid methyl esters (FAMEs), by API 20E profiling and/or by Fluorescent Amplified Fragment Length Polymorphism (FAFLP) DNA fingerprinting. In general, Ot(r) hospital heterotrophs displayed a higher frequency (84%) of ampicillin (Amp) tolerance compared to the Ot(r) heterotrophs from the freshwater fishfarm site (22%). FAME results indicated that this effect was linked to the predominance of intrinsically ampicillin-resistant Ot(r) Aeromonas strains over representatives of Acinetobacter and Escherichia coli within the hospital strain set. Among the Ot(r) mesophilic Aeromonas strain set, the global tolerance profiles of the two sites only differed in a higher number of kanamycin (Kan) -tolerant strains (43%) for hospital aeromonads in comparison with the fishfarm aeromonads (8%). To some extent, this finding was correlated with the specific presence of Aeromonas caviae DNA hybridisation group (HG) 4. Collectively, these results suggest that the profiles for Amp and Kan tolerance observed in both sites arose from taxonomic differences in the culturable Ot(r) bacterial population at the generic or subgeneric level. In addition, our identification data also revealed that Enterobacter sp., Stenotrophomonas maltophilia, and A. veronii biovar sobria HG8 may be considered potential indicator organisms to assess microbial tolerance in various compartments of the aquatic environment. PMID- 11403392 TI - A new sterigmatocystin-producing Emericella variant from agricultural desert soils. AB - An unusual, sterigmatocystin-producing taxon with characteristics of both Emericella nidulans (anamorph Aspergillus nidulans) and Emericella rugulosa (anamorph Aspergillus rugulovalvus, formerly A. rugulosus) was isolated repeatedly during a mycofloral survey of desert cotton field soils where aflatoxin is a chronic problem. Members of this taxon had ascospores with smooth convex walls like E. nidulans but grew slowly like E. rugulosa; moreover, they were similar to an industrial echinocandin B-producing strain which had been classified as "Aspergillus nidulans var. roseus." These new desert isolates were compared with "A. nidulans var. roseus" and representative wild-type isolates of E. nidulans and E. rugulosa using traditional morphological characters, secondary metabolite profiles of mycelial extracts, and Southern blot analysis of genomic DNA. The desert isolates and "A. nidulans var. roseus shared morphological, physiological and molecular characters with E. rugulosa. These isolates constitute a new non-rugulose variant of E. rugulosa. PMID- 11403393 TI - Fusobacterium prausnitzii and related species represent a dominant group within the human fecal flora. AB - The human gut microflora plays a key role in nutrition and health. It has been extensively studied by conventional culture techniques. However these methods are difficult, time consuming and their results not always consistent. Furthermore microscopic counts indicate that only 20 to 40% of the total flora can be cultivated. Among the predominant species of the human gut, Fusobacterium prausnitzii was reported either as one of the most frequent and numerous species or was seldom retrieved. We designed and validated a specific rRNA-targeted oligonucleotide probe, called S-*-Fprau-0645-a-A-23, to accurately detect and quantify F. prausnitzii and relatives within the human fecal microflora. The target group accounted for 5.3 +/- 3% of total bacterial 16S rRNA using dot blot hybridization (10 human fecal samples) and 16.5 +/- 7% of cells stained with Dapi using in situ hybridization (10 other human fecal samples). A specific morphology seemed to be typical and dominant: two cells forming an asymmetrical double droplet. This work showed that F. prausnitzii and phylogenetically related species represent a dominant group within the human fecal flora. PMID- 11403394 TI - Gene transfer systems and their applications in Archaea. AB - Members of the Archaea domain are extremely diverse in their adaptation to extreme environments, yet also widespread in "normal" habitats. Altogether, among the best characterized archaeal representatives all mechanisms of gene transfer such as transduction, conjugation, and transformation have been discovered, as briefly reviewed here. For some halophiles and mesophilic methanogens, usable genetic tools were developed for in vivo studies. However, on an individual basis no single organism has evolved into the "E. coli of Archaea" as far as genetics is concerned. Currently, and unfortunately, most of the genome sequences available are those of microorganisms which are either not amenable to gene transfer or not among the most promising candidates for genetic studies. PMID- 11403395 TI - Structural investigations on the cell surface of Erysipelothrix rhusiopathiae. AB - The cell wall of Erysipelothrix rhusiopathiae was investigated. The corrected structure of the murein, which was published earlier, is reported here. The murein belongs to the B1delta type. It has L-alanine in position three of the peptide subunit and is crosslinked via a glycine-L-lysine-L-lysine interpeptide bridge. As expected by the complex serological situation in E. rhusiopathiae the carbohydrate moiety of the cell wall proofed to be very intricate. As accessory polymers polysaccharide(s) and teichoic acid(s) were identified. The teichoic acid(s) possess two novel polyols of which one could not be identified, the other one behaved like 6-O-methylgalactitol in gas liquid chromatography. Glucose and N acetylgalactosamine were determined as substituents. The polysaccharide(s) is composed of mainly N-acetylfucosamine and smaller amounts of galactose, N acetylglucosamine, and a further unidentified sugar appearing later than hexaacetylsorbitol in GLC. PMID- 11403396 TI - Effects of deflected droplet electrostatic cell sorting on the viability and exoproteolytic activity of bacterial cultures and marine bacterioplankton. AB - The cell-sorting capability of flow cytometers makes it possible to isolate specific populations of cells with pre-defined cytometric characteristics. A better knowledge of the biological effects of the sorting process is necessary for the future cell sorting applications. In this paper we report the effects of flow cytometric sorting on bacterial viability and exoproteolytic activity (EPA) of bacterial cultures and marine bacterioplankton. Sorting bacterial cultures and bacterioplankton samples reduce viability as assessed by plate counts and produce variations in the exoproteolytic activity. These effects indicate that deflected electrostatic sorting may significantly alter the biological properties of the sorted bacteria. PMID- 11403397 TI - Alcaligenes faecalis subsp. parafaecalis subsp. nov., a bacterium accumulating poly-beta-hydroxybutyrate from acetone-butanol bioprocess residues. AB - The authors have previously isolated a solvent tolerant bacterium, strain G(T), (T = type strain) capable to convert acetone-butanol bioprocess residues into poly-beta-hydroxybutyrate. Strain G(T) was initially identified as Alcaligenes spp by standard bacteriological tests. In this study the taxonomic position of the bacterium was investigated in detail. The 165 rDNA sequence analysis, the G + C content of DNA (56 mol%) and the presence of ubiquinone Q-8 confirmed strain G(T) as a representative of the genus Alcaligenes. In the polyamine pattern of the bacterium putrescine and cadaverine were detected, but only trace amounts of 2-hydroxyputrescine. The extremely low content of 2-hydroxyputrescine is remarkable, since this unique diamine is a common marker for beta-proteobacteria. Phylogenetic analyses of 16S rDNA demonstrated that Alcaligenes sp. G(T) is most closely related to the species Alcaligenes faecalis (99.6% sequence similarity to A. faecalis HR4 and 98.7% sequence similarity to A. faecalis [ATCC 8750T = DSM 30030T]. On the basis of DNA-DNA relatedness (56% similarity), the unique polyamine pattern, the physiological and biochemical differences strain G(T) could be distinguished from the species A. faecalis. Therefore, a new subspecies for the species Alcaligenes faecalis is proposed; Alcaligenes faecalis subsp. parafaecalis subsp. nov. PMID- 11403398 TI - Psychrobacter proteolyticus sp. nov., a psychrotrophic, halotolerant bacterium isolated from the Antarctic krill Euphausia superba Dana, excreting a cold adapted metalloprotease. AB - An Antarctic marine bacterium (strain 116) excreting an extracellular cold adapted metalloprotease was subjected to a detailed polyphasic taxonomic investigation. Strain 116 was previously isolated from the stomach of a specimen of the Antarctic krill Euphasia superba Dana and tentatively characterized as Sphingomonas paucimobilis 116. The 16S rDNA sequence analysis showed that the strain is in fact related to species of the genus Psychrobacter, next to Psychrobacter glacincola (97.4% similarity). Sequence similarities between strain 116 and other Psychrobacter species ranged from 96.9% (with P. urativorans) to 95.4% (with P. immobilis). Key phenotypic characteristics as well as chemotaxonomic features of the bacterium were congruent with the description of the genus Psychrobacter i.e. cells were strictly aerobic, strongly oxidase positive, psychrotrophic, halotolerant, gram-negative non-motile coccobacilli, with ubiquinone-8 as the main respiratory lipoquinone and 18:1 cis 9, 16:1 cis and 17:1 (omega8c being the predominant cellular fatty acids. The G+C content of the DNA was 43.6 mol%. DNA-DNA hybridization studies showed that the relatedness between strain 116 and Psychrobacter glacinola is only 62.2%. Further differences were apparent in whole-cell SDS-PAGE protein pattern, cellular fatty acid profile and in a number of physiological and biochemical characteristics as well as in enzymatic activities. Tolerance to 5% bile salts, nitrate reduction, citrate utilization, acid production from carbohydrates, alkaline phosphatase, acid phosphatase, C4 esterase, C14 lipase and valine arylamidase were found to differentiate strain 116 from Psychrobacter glacincola. On the basis of this phenotypic and molecular evidences, strain 116, previously known as Sphingomonas paucimobilis 116, was recognized as a new species of the genus Psychrobacter for which the name Psychrobacter proteolyticus is proposed. Strain 116 has been deposited in the Collection de l'Institut Pasteur, France, as CIP106830T and in the Deutsche Sammlung von Mikroorganismen and Zellkulturen, as DSM13887. PMID- 11403399 TI - Diversity of polyamine patterns in soft rot pathogens and other plant-associated members of the Enterobacteriaceae. AB - Polyamine profiles of 91 pectolytic and other plant-associated strains from 30 taxa of the Enterobacteriaceae were obtained by gradient high performance liquid chromatography (HPLC). Pectobacterium carotovorum, basonym Erwinia carotovora, contained a high amount of putrescine and less diaminopropane. Diaminopropane was absent in Pectobacterium chrysanthemi, basonym E. chrysanthemi, whereas cadaverine was present in addition to the major compound putrescine. This chemotaxonomic difference reflects the deepest phylogenetic branching point within the recently emended genus Pectobacterium which lies between the two species P. carotovorum and P. chrysanthemi. Both important soft rot pathogens are easily distinguishable from each other and from the type species of the genus Erwinia as diaminopropane is the only major polyamine compound in E. amylovora. Chemotaxonomic heterogeneity is also emerging with respect to DYE's Amylovora group proposed in an early phytopathological concept. PMID- 11403400 TI - Genetic characterization of a Chlamydophila pneumoniae isolate from an African frog and comparison to currently accepted biovars. AB - The amphibian isolate DE177 identified as Chlamydophila (C.) pneumoniae was sequenced in five genomic regions: 16S ribosomal RNA gene, 16-23S intergenic spacer, ompA, ompB, and groESL genes. Comparison with corresponding sequences of the currently accepted equine, human and koala biovars of C. pneumoniae revealed that koala strains represented the most closely related taxon, although sequence dissimilarities in the ompA (VD4) and ompB gene regions were noted. In this respect, the present isolate is distinct from a previously described frog isolate (Berger et al., 1999) whose sequence analysis yielded identity to the koala biovar. As three of the nucleotide substitutions in ompA (VD4) of DE177 will be translated into two altered amino acids the possible existence of another biovar is discussed. PMID- 11403401 TI - Distinct electrophoretic isozyme profiles of Fusarium graminearum and closely related species. AB - Cellulose-acetate electrophoresis (CAE) was used to investigate isozyme polymorphisms among different isolates of Fusarium cerealis, F. culmorum, F. graminearum and F. pseudograminearum from around the world. After initial testing of 22 enzymes in three buffer systems for activity and resolution of bands, 12 proved to be appropriate for analysis of the full sample set. Remarkably uniform isozyme patterns were obtained intraspecifically, irrespective of the geographical origin of the isolates or the host/substratum from which they were isolated. This result indicated that isolates within a given species are descendant from a same ancestral population. Comparing the different electrophoretic types (ETs), adenylate kinase (AK), NADP dependent glutamate dehydrogenase (NADP GDH), peptidase B (PEP B), peptidase D (PEP ID) and phosphoglucomutase (PGM) proved to be diagnostic for at least one species examined. However, only PEP D was useful alone as a marker to distinguish the four taxa studied providing a rapid and simple CAE based diagnostic protocol. Cluster analysis of band sharing coefficients grouped the isolates into four distinct groups corresponding to the 4 species studied. Isolates of F. cerealis were clustered between those of F. culmorum and F. graminearum corroborating their known close relationship to both species. For common ETs, the similarity values between F. cerealis and F. culmorum and between F. cerealis and F. graminearum were the same. Furthermore, the similarity values and the resulting phenogram indicated that F. graminearum is more closely related to F. cerealis and F. culmorum than to F. pseudograminearum, thus the morphological similarity of F. graminearum and F. pseudograminearum does not reflect their generic relationship. This fact supports the species status of F. pseudograminearum. PMID- 11403402 TI - Development of 18S rRNA-targeted oligonucleotide probes for specific detection of Hartmannella and Naegleria in Legionella-positive environmental samples. AB - Aquatic protozoa are natural hosts of the human pathogen Legionella pneumophila. The fluorescence labeled 16S rRNA-targeted oligonucleotide probe LEGPNE1 has recently been shown to specifically detect extracellular legionellae as well as intracellular legionellae parasitizing protozoa. In this study we designed oligonucleotide probes which are complementary to distinct regions of the 18S rRNA of the Legionella host organisms of the genera Hartmannella and Naegleria. The specificity of the probes, HART498 and NAEG1088, was tested by in situ hybridization of various laboratory reference strains. In order to evaluate the fluorescent probes for environmental studies three selected Legionella-positive cold water habitats were examined for the presence of these protozoa. Traditional culture methods followed by morphological identification revealed an almost consistent presence of Naegleria spp. in cold water habitats. Other protozoa species including Acanthamoeba spp., Echinamoeba spp., Hartmannella spp., Platyamoeba placida, Saccamoeba spp., Thecamoeba quadrilineata, and Vexillifera spp. were found sporadically. Concomitant analysis of the pH, conductivity and temperature of the water samples revealed no preference of Legionella or the respective protozoa for certain environmental conditions. The specificity of the newly designed 18S rRNA probes demonstrates that they are valuable and rapid tools for the identification of culturable environmental protozoa. PMID- 11403403 TI - Design and application of new 16S rRNA-targeted oligonucleotide probes for the Azospirillum-Skermanella-Rhodocista-cluster. AB - The genera Azospirillum, Skermanella and Rhodocista form a phylogenetic subgroup within the alfa subclass of Proteobacteria. Based on comparative 16S rRNA sequence analysis a nested set of new oligonucleotide probes was designed. It comprises probes for the whole genus cluster Azospirillum-Skermanella-Rhodocista, for the Azospirilli subcluster I including A. lipoferum, A. doebereinerae, A. largimobile, A. brasilense and A. halopraeferens, for the Azospirilli subcluster II including A. amazonense, A. irakense and the genus Skermanella, for the genus Rhodocista as well as for all Azospirilli species or species cluster. The new probes allow a fast and reliable in situ identification of bacteria belonging to the Azospirillum-Skermanella-Rhodocista-cluster at different phylogenetic levels. The specificity of the new probes was tested with 56 strains of the Azospirillum Rhodocista-Skermanella-cluster and selected reference cells from other genera by hybridising with the complete probe set. In addition, applications of the fluorescently labelled probes for in situ identification of isolates and for the in situ localisation of A. brasilense on maize roots were demonstrated using confocal laser scanning microscopy. PMID- 11403405 TI - Nosocomial infections. PMID- 11403404 TI - Characterization of facultative oligotrophic bacteria from polar seas by analysis of their fatty acids and 16S rDNA sequences. AB - One hundred and seventy three bacterial strains, isolated previously after enrichment under oligotrophic, psychrophylic conditions from Arctic (98 strains) and Antarctic seawater (75 strains), were characterized by gas-liquid chromatographic analysis of their fatty acid compositions. By numerical analysis, 8 clusters, containing 2 to 59 strains, could be delineated, and 8 strains formed separate branches. Five clusters contained strains from both poles, two minor clusters were confined to Arctic isolates, and one cluster consisted of Antarctic isolates only. The 16S rRNA genes from 23 strains, representing the different fatty acid profile clusters and including the unclustered strains, were sequenced. The sequences grouped with the alpha and gamma Proteobacteria, the high percent G+C gram positives, and the Cytophaga-Flavobacterium-Bacteroides branch. The sequences of strains from 4 clusters and of 7 unclustered strains were closely related (sequence similarities above 97%) to reference sequences of Sulfitobacter mediterraneus, Halomonas variabilis, Alteromonas macleodii, Pseudoalteromonas species, Shewanella frigidimarina, and Rhodococcus fascians. Strains from the other four clusters and an unclustered strain showed sequence similarities below 97% with nearest named neighbours, including Rhizobium, Glaciecola, Pseudomonas, Alteromonas macleodii and Cytophaga marinoflava, indicating that the clusters which they represent form as yet unnamed taxa. PMID- 11403406 TI - Distribution and expression of the astA gene (EAST1 toxin) in Escherichia coli and Salmonella. AB - The distribution and expression of the astA gene (EAST1 toxin) among 358 strains of Enterobacteriaceae were studied. The gene was found in 32.6% and 11.9% of Escherichia coli and Salmonella strains, respectively. The majority of E. coli EAST1-positive strains were found among EHEC (88.0%), EAggEC (86.6%), and A-EPEC (58.3%). The gene was present in 16.6% of E. coli strains without known virulence genes. There was no significant variation among the different serotypes of E. coli tested regarding the presence of the gene. For EPEC, 13.7% of the tested strains were astA-positive. Among atypical EPEC (eae+, bfp-, EAF-) and (eae+, bfp+, EAF-) 46.2 and 72.7%, respectively, were positive. The majority of the A EPEC (87%) and EaggEC (83%) strains expressed the EAST-1 toxin as judged from Ussing chamber experiments. Of 32 EIEC strains studied, 2 possessed and expressed the gene as determined in Ussing chamber experiments. Among the Salmonella strains studied, five strains isolated from food were positive for astA and one strain of S. agona showed biological activity in Ussing chamber experiments. PMID- 11403407 TI - Investigation of infection with Campylobacter jejuni in a man with hypogammaglobulinaemia using PCR-single-stranded conformational polymorphism (PCR SSCP) typing. AB - This study investigated several episodes of infection of Campylobacter jejuni in an immunocompromised male with hypogammaglobulinaemia, presenting with diarrhoea and bacteraemia over a 16-month period, by employing three phenotyping and four genotyping schemes, including the single-stranded conformational polymorphism (SSCP) technique to establish if infection was reinfection or persistent infection. Four isolates from blood culture and two faecal isolates of Campylobacter jejuni were obtained from the patient by direct selective plating on Skirrow Selective agar. Isolates were characterised at the sub-species level by Penner serology, Preston biotyping, Preston phage-typing, as well as E3CJC2 restriction fragment length polymorphism (RFLP), 16S ribotyping, flagellin (flaA) RFLP and single-stranded conformational polymorphism (SSCP) analyses. Phenotyping and genotyping sub-species analyses demonstrated that the patient was infected with at least two different strains of Campylobacter jejuni, i. e. one strain that persisted throughout the 16-month period and another strain that was transient suggesting reinfection from a different source. SSCP analysis was the most discriminatory of all the typing schemes examined and demonstrated an altered genotype of the persistent strain, whereby there were subtle modifications to the hypervariable regions of the flaA gene. Overall, as SSCP examines the hypervariable region of the flaA gene and as this technique can detect point mutations, differences between SSCP banding patterns may represent markers and thus examine mutations that occur under immune selection, thereby permitting the C. jejuni to evade the host immune response. In conclusion, this study describes the novel use of SSCP genotyping of C. jejuni and demonstrated that this method is a highly discriminatory technique which may be beneficial in outbreak characterisation, but which is not suitable to examine the clonal patterns of C. jejuni over a long period of time. PMID- 11403408 TI - Neutralisation of cytotoxic vacuolating activity by serum antibodies of Helicobacter pylori-infected patients. AB - The study involved 196 H. pylori strains and 196 serum samples taken from the same patients. H. pylori strains were investigated for the production of vacuolating cytotoxin. Antibodies to the vacuolating cytotoxin produced by H. pylori were detected in the sera samples by neutralisation assay (on Intestine 407 cells) and ELISA. Of the 196 H. pylori strains tested, 80 (40.8%) were found to express vacuolating cytotoxic activity. The titres of cytotoxic nonconcentrated broth culture filtrates ranged from 1:2 to 1:128. The vacuolating assay was positive in 37.1% strains isolated from children, and in 50% strains isolated from adults. Cytotoxin-positive H. pylori strains were found more frequently in duodenal ulcer (71%) than in chronic gastritis (35.2%) patients, and this difference was statistically significant p < 0.05. Neutralising antibodies to vacuolating cytotoxin were present in 51% and 49% of the serum samples tested by neutralisation and ELISA, respectively. Duodenal ulcer patients had antibodies to vacuolating cytotoxin more frequently (p < 0.05) than chronic gastritis patients. Antibodies to cytotoxin were detected in the serum samples from patients infected by cytotoxic (100%) and noncytotoxic (18%) H. pylori strains. PMID- 11403409 TI - Zentralblatt fur Bakteriologie--100 years ago: a dispute about priority of forensic blood test. PMID- 11403410 TI - Experimental Helicobacter pylori infection of rhesus macaques (Macaca mulatta). AB - The aim of the present study was to establish an animal model for Helicobacter pylori (H. pylori) infection at the German Primate Centre in rhesus monkeys (Macaca mulatta). During the experiments the susceptibility of three animals to different H. pylori strains of human origin was tested. In a follow-up study gastric biopsies from three different sites were investigated in regular intervals using microbiological, histological, electron microscopical and molecular biological methods to evaluate the presence of bacterial colonization and the occurrence of gastritis. It was possible to establish a persistent experimental infection. The rather long follow-up period of 18 months offered the possibility to demonstrate a permanent H. pylori infection in the gastric mucosa of the test animals. The three animals have now been successfully colonized with H. pylori for 18 months and presented a chronic active gastritis confirmed by microbiological and histological methods. By molecular typing, the identity of the isolates recovered from the animals was shown. It was possible to demonstrate that one infection strain outcompeted the second one. Taken together, prerequisites exist for making use of an attractive and useful animal model in rhesus monkeys especially for long term observations. PMID- 11403411 TI - Virulence of group A streptococci in fertile hens eggs is mainly effected by M protein and streptolysin O. AB - In this study we have investigated whether streptolysin O contributes to the virulence of group A streptococci. For this purpose we generated M-negative and SLO-negative mutants by insertion mutagenesis into the chromosome of an M type 1 strain. The inactivation of M1 protein expression was achieved by the construction of the integrative plasmid pSFABS, which contains the internal fragment abs of the emm1 gene. Integration of pSFABS by homologous recombination into the chromosome of strain 38 541 resulted in the generation of mutant EMM1. Inactivation of slo with plasmid pFWSLOD resulted in two different mutant forms. The homologous recombination with plasmid pFWSLOD carrying the two slo fragments slo1 (899 base pairs in the 5' region) and slo2 (709 base pairs in the downstream part) resulted in mutants SLO3, SLO4 and SLO17. In SLO17 a double crossover event took place with insertion of the spectinomycin resistance gene aad9 between the slo fragments 1 and 2. In mutants SLO3 and SLO4 the homologous recombination with the same plasmid led to the integration of the whole plasmid construct into the chromosome of strain 38 541. Both forms of mutation failed to express SLO. In mutant SLO4 additionally M1 protein expression was significantly decreased. The mutants EMM1 (M-, SLO+) and SLO4 (M decreased, SLO-) showed a reduced binding to collagen-coated surfaces. In contrast the mutants SLO3 and SLO17 (both M+, SLO-) and the wild-type strain 38 541 (M+, SLO+) showed an affinity to collagen similar to purified M1 protein. All mutants were less virulent for chicken embryos compared to the wild-type strain after infection by intravenous injection as well as by application onto the chorioallantoic membrane. The results show that besides M protein SLO can also influence virulence of group A streptococci. Moreover, it became obvious that streptococci need more than one tool to fully develop their infectious potential. PMID- 11403412 TI - Subtyping of MRSA isolates belonging to a widely disseminated clonal group by polymorphism of the dru sequences in mec-associated DNA. AB - During the past 7 years the "Berlin epidemic MRSA" has spread throughout whole Germany. SmaI macrorestriction patterns of this clonal group are rather stable as are the length polymorphisms at the 3' end of the coagulase gene and the x-region of spa. However, the dru region (direct repeat units) of mec-associated DNA exhibits a length polymorphism due to deletion of one or more direct repeat units. Five different subclones could be discriminated by dru region length polymorphism. Location of deletions and of a few point mutations allow a delineation of these subclones. PMID- 11403413 TI - Adhesins as invasins. PMID- 11403414 TI - Protein kinases and kinase-modulated effectors in the late phase of ischemic preconditioning. AB - In the 15 years since the first publication on ischemic preconditioning (PC), our knowledge of this phenomenon has increased exponentially. While the original studies described the early phase of ischemic PC, we now know that a second or late phase of ischemic PC also exists. In particular, the late phase of PC is triggered by factors such as adenosine, opioids, radicals, and nitric oxide. These factors in turn initiate a molecular chain reaction, which includes activation of serine/threonine kinases, tyrosine kinases, and mitogen-activated protein kinases. Subsequently, this cascade of reactions modulates a plethora of cardioprotective proteins including heat shock proteins, KATP channels, nitric oxide synthase, cyclooxygenase-2, and antioxidants. However, despite this phenomenal amount of information, the construction of a unifying hypothesis describing the signaling mechanism of late PC has proved challenging. The purpose of this article, therefore, is to review the current literature and hypothesis regarding the signaling system in the late phase of ischemic PC, to tackle areas of controversy within this model, and to address potential future directions of investigation that will hopefully promote the generation of a unifying paradigm. PMID- 11403415 TI - The role of apoptosis in myocardial ischemia: a critical appraisal. AB - The role of apoptosis in cardiac ischemia is not clarified yet. Own data show that suicidal cell death is apparently not important in global ischemia where it only affects a small number of myocytes (8 %) while the majority of cells, i.e. 92 % die by oncosis. In acute regional ischemia it is most probably not a decisive factor. However, more solid data are needed to justify this statement. Human hibernating myocardium shows an activation of the apoptotic cascade, i.e., apoptosis might contribute to cell loss in this pathophysiological situation of multiple ischemic episodes. Manifold unresolved issues contribute to problems in determining the role of apoptosis in ischemia. These include 1) Uncertainty of the duration of the apoptotic cascade from activation of death receptors at the cellular membrane until DNA fragmentation occurs, 2) The role of the mitochondrial pathway, 3) The mode of removal of myocytes after cell death has occurred, 4) Technical problems such as specificity of the TUNEL method, detection of low abundance proteins such as activated caspases or cytochrome C, statistical considerations. These issues and many others should be clarified before any definite conclusion as to the role of apoptosis in ischemia may be drawn. PMID- 11403416 TI - In vivo myocardial gene transfer: optimization, evaluation and direct comparison of gene transfer vectors. AB - The purpose was to determine the relative efficiency, toxicity and duration of expression following gene delivery by intramyocardial injection of naked DNA, naked DNA complexed to cationic liposomes, naked DNA complexed to cationic liposomes with integrin-targetting peptide, recombinant (E1-/E3-) adenovirus, recombinant adeno-associated virus and recombinant (ICP27-) herpes simplex virus. All vectors incorporated a LacZ reporter driven by a promoter containing the hCMV IE promoter/enhancer. Efficiency was scored by counting positive cells in five standard microscopic sections harvested from the left ventricular apex. Rabbit hearts (n = 100) were examined from 2 to 56 days after injection. Uncomplexed and complexed naked DNA were very inefficient with less than one positive cell visible per heart. The viral vectors all resulted in robust gene expression with adenovirus being the most efficient by at least one order of magnitude before 21 days. However, despite disparate titres, the efficiency beyond 21 days of adenovirus and adeno-associated virus were comparable. In contrast to adeno associated virus, both adenovirus and herpes-simplex virus were associated with a marked inflammatory response. Despite reporter gene activity appearing only after 21 days, adeno-associated virus shows comparative promise as a myocardial gene delivery vector. PMID- 11403417 TI - Electrophysiological characterization of murine myocardial ischemia and infarction. AB - BACKGROUND: Genetically altered mice will provide important insights into a wide variety of processes in cardiovascular physiology underlying myocardial infarction (MI). Comprehensive and accurate analyses of cardiac function in murine models require implementation of the most appropriate techniques and experimental protocols. OBJECTIVE: In this study we present in vivo, whole-animal techniques and experimental protocols for detailed electrophysiological characterization in a mouse model of myocardial ischemia and infarction. METHODS: FVB mice underwent open-chest surgery for ligation of the left anterior descending coronary artery or sham-operation. By means of echocardiographic imaging, electrocardiography, intracardiac electrophysiology study, and conscious telemetric ECG recording for heart rate variability (HRV) analysis, we evaluated ischemic and post-infarct cardiovascular morphology and function in mice. RESULTS: Coronary artery ligation resulted in antero-apical infarction of the left ventricular wall. MI mice showed decreased cardiac function by echocardiography, infarct-typical pattern on ECG, and increased arrhythmia vulnerability during electrophysiological study. Electrophysiological properties were determined comprehensively, but were not altered significantly as a consequence of MI. Autonomic nervous system function, measured by indices of HRV, did not appear altered in mice during ischemia or infarction. CONCLUSIONS: Cardiac conduction, refractoriness, and heart rate variability appear to remain preserved in a murine model of myocardial ischemia and infarction. Myocardial infarction may increase vulnerability to inducible ventricular tachycardia and atrial fibrillation, similarly to EPS findings in humans. These data may be of value as a reference for comparison with mutant murine models necessitating ischemia or scar to elicit an identifiable phenotype. The limitations of directly extrapolating murine cardiac electrophysiology data to conditions in humans need to be considered. PMID- 11403418 TI - Simultaneous in utero assessment of AV nodal and ventricular electrophysiologic parameters in the fetal sheep heart. AB - BACKGROUND: Fetal tachyarrhythmias are usually of supraventricular origin. To investigate whether specific electrophysiologic properties of the fetal heart contribute to this preponderance by either favoring supraventricular tachycardias or by rendering ventricular tachycardias unlikely, we measured fetal electrophysiologic parameters in utero using transuterine fetal transesophageal electrocardiograms in fetal sheep. Since overdrive pacing may help to establish the mechanism of an arrhythmia and may be used to treat fetal tachycardias, different modes of transesophageal pacing in utero were also assessed. METHODS AND RESULTS: Decapolar electrophysiology catheters were fetoscopically inserted into the esophagus of 9 fetal sheep (pregnancy duration 94- 105 days, term = 145 days). Electrocardiograms were recorded simultaneously from all adjacent bipoles and from two pacing wires sutured onto the fetal shoulders. Pacing was attempted either via two adjacent electrodes of the intraesophageal catheter or via the most distal and most proximal electrode. Fetal cycle length, PQ, and QT intervals were close to (approx. 75 %), but fetal QRS duration was < 20 % of maternal values, thus shifting the relation between activation and repolarization towards longer excitation wave lengths. Fetal QT dispersion was small (< or = 10 ms). Atrial pacing was achieved in all fetuses using distant electrodes, and with lower thresholds when compared to closely spaced bipolar electrodes (p < 0.05). CONCLUSIONS: (I) An altered relation between ventricular activation and repolarization and a low dispersion of ventricular repolarization may protect the fetal heart against ventricular reentrant tachycardias. (II) Relatively normal fetal AV nodal conduction delay already provides one of the prerequisites for supraventricular reentrant tachycardias involving the AV node at this stage of fetal development. (III) High-rate esophageal pacing of the fetal atria is best achieved using widely spaced bipolar pacing electrodes. PMID- 11403419 TI - Chronic effects of ACE-inhibition (quinapril) and angiotensin-II-type-1 receptor blockade (losartan) on atrial natriuretic peptide in brain nuclei of rats with experimental myocardial infarction. AB - Alterations of the central nervous system may be important for imbalance of cardiovascular and fluid regulation in heart failure. The central renin angiotensin and atrial natriuretic peptide (ANP) systems act as mutual antagonists. The effects of angiotensin converting enzyme (ACE) inhibition (quinapril, 6 mg/kg/day) and angiotensin II type 1 (AT1) receptor blockade (losartan, 10 mg/kg/day) on ANP levels in 18 selected, microdissected brain nuclei were determined in sham-operated rats and rats with left ventricular dysfunction 8 weeks after myocardial infarction (MI). Plasma ANP tended to increase in MI rats and was further increased by quinapril. ANP was decreased in 12 brain areas of MI rats. ANP concentration was also significantly decreased by quinapril in six brain nuclei including subfornical organ and organum vasculosum laminae terminalis (areas lacking blood-brain barrier), and by losartan in 16 brain nuclei outside and within the blood-brain barrier in sham operated rats. However, both quinapril and losartan prevented a further reduction of central ANP as a result of myocardial infarction. These data suggest that there are effects on central ANP that result from chronic left ventricular dysfunction as well as an ACE-inhibitor and AT1-antagonist. Mechanisms and consequences of central ANP depression remain unclear. They could, however, support systemic vasoconstriction and sodium and fluid retention. PMID- 11403420 TI - Production and metabolism of ceramide in normal and ischemic-reperfused myocardium of rats. AB - Ceramide has been shown to be a key signaling molecule involved in the apoptotic effect of tumor necrosis factor alpha (TNF-alpha) and other cytokines. Given the importance of cytokines such as TNF-alpha in myocardial ischemia-reperfusion injury, we hypothesize that ceramide is increased during ischemia or reperfusion, and that the activity of enzymes responsible for its production or breakdown should be increased and/or decreased, respectively. Therefore, in the present study, we characterized the enzymatic activities responsible for ceramide production and metabolism in the myocardium of rats, and determined the contribution of these enzymes to altered ceramide levels during myocardial ischemia and reperfusion. The basal ceramide concentration in the myocardium of rats was 34.0 pmol/mg tissue. As determined by the conversion of 14C sphingomyelin into ceramide and 14C-choline phosphate, both neutral (N-) and acidic (A-) SMase were detected in the myocardium, with a conversion rate of 0.09 +/- 0.008 and 0.32 +/- 0.05 nmol/min per mg protein, respectively. The activity of A-SMase (78 % of total cellular activity) was significantly higher in microsomes than in cytosol, while the activity of N-SMase was similar in both fractions. Ceramidase, a ceramide-metabolizing enzyme, was also detected in the myocardium of rats. It metabolized ceramide into sphingosine at a rate of 9.94 +/ 0.42 pmol/min per mg protein. In anesthetized rats, 30 min of ischemia had no apparent effect on ceramide concentrations in the myocardium, while 30 min of ischemia followed by 3 h of reperfusion resulted in a significant increase in ceramide by 48 %. The activities of both N- and A-SMase were reduced by 44 % and 32 %, respectively, in the myocardium subjected to ischemia followed by reperfusion, but unaltered in the ischemic myocardium. It was also found that myocardial ischemia followed by reperfusion produced a marked inhibition of ceramidase (by 29 %). These results demonstrate that the myocardium of rats expresses N- and A-SMase and ceramidase, which contribute to the production and metabolism of ceramide, respectively. Tissue ceramide concentrations increased in reperfused myocardium. These increases in ceramide were not associated with enhanced SMase activity, but rather with reduced ceramidase activity. PMID- 11403421 TI - Inhibition of smooth muscle cell proliferation after local drug delivery of the antimitotic drug paclitaxel using a porous balloon catheter. AB - Percutaneous transluminal coronary angioplasty is an accepted treatment for coronary artery disease. The major limitation, however, is the high incidence of restenosis which limits the long-term benefit of this intervention. Paclitaxel is a new antiproliferative agent that has generated considerable scientific interest since it was introduced in clinical trials in the early 1980s. Recent in vitro studies have shown that paclitaxel has considerable antiproliferative activity in human coculture systems. In the present study the efficacy of paclitaxel was investigated after development of an intimal plaque by electrical stimulation and additional cholesterol diet and subsequent balloon angioplasty in 63 New Zealand White rabbits. Local drug delivery of paclitaxel was accomplished in 30 rabbits with a porous balloon catheter (35 holes, hole diameter 75 microm, 2.5 mm catheter diameter). Paclitaxel was administered locally with 4 ml (solution 10( 5) mol/L) using an injection pressure of 2 atm. To study the extent of restenosis and morphological changes, the animals were sacrificed 7, 28 or 56 days after intervention. After staining procedures quantification of SMC proliferation, intimal macrophages and morphological analyses were performed. Paclitaxel plasma concentrations were measured using HPLC technique. One week after balloon angioplasty the arteries treated with local paclitaxel delivery showed an insignificant trend towards a reduction in intimal smooth muscle cell proliferation (untreated 8.4 +/- 4.9 % vs paclitaxel treated 2.4 +/- 2.4 %, p = NS). However, this resulted in a significant reduction of stenosis degree of 66 % 8 weeks after intervention compared to the untreated group (untreated 41 +/- 18 % vs paclitaxel treated 14 +/- 11 %, p = 0.005). In conclusion, locally delivered paclitaxel prevented neointimal thickening in the rabbit carotid artery after balloon angioplasty. Local paclitaxel treatment may therefore be a clinical option for the prevention of restenosis after coronary interventions. However, further preclinical studies have to prove long-term efficacy and safety. PMID- 11403422 TI - Lovastatin controls signal transduction in vascular smooth muscle cells by modulating phosphorylation levels of mevalonate-independent pathways. AB - Lovastatin has been proven to effectively lower circulating LDL cholesterol and to exert antiproliferative effects on various cell lines, the latter effect being only incompletely understood. We found that lovastatin modulates the signal transducing phosphorylation cascade in vascular smooth muscle cells in a mevalonate-independent manner. Lovastatin was found to distinctively increase total phosphotyrosine levels in smooth muscle cells, an effect which could not be restored by mevalonate. At a concentration of 5 micromol/L lovastatin had a highly specific effect on the mitogen-activated protein kinase pathway. The expression of p42/44 mitogen-activated protein kinase (MAPK) was clearly reduced, but could be restored by addition of mevalonate, while the phosphorylation of p44 was mildly suppressed and the phosphorylation of p42 MAPK was reduced to non detectable levels. While the phosphorylation of p44 MAPK could partially be restored by addition of mevalonate, the reduced phosphorylation of p42 MAPK could not be restored by addition of excessive doses of mevalonate or stimulation of the cells with basic fibroblast growth factor. Concurrently the expression of the GTP-binding Ras protein was significantly elevated at 5 and 20 micromol/L lovastatin, this effect being attenuated by addition of mevalonate to cell cultures. The data indicate that lovastatin is capable of modulating cellular signaling independently of the cholesterol synthesis pathway. PMID- 11403423 TI - Impaired beta-adrenergic control of immune function in patients with chronic heart failure: reversal by beta1-blocker treatment. AB - BACKGROUND: In patients with chronic heart failure (CHF) and heightened sympathetic activity alterations in immune function have been described. OBJECTIVES: To find out whether, in CHF patients, beta-blocker treatment might beneficially affect immune function. METHODS: We studied activation of circulating lymphocytes (assessed as concanavalin A (CON A)-induced inositol phosphate (IP) formation and proliferation ([3H]-thymidine incorporation) from 8 CHF patients on standard medication (Group A, mean age 54 +/- 6 yrs, NYHA class II - IV, mean 3.1 +/- 0.3) and in 9 CHF patients on standard medication and additional treatment with the beta1-blocker metoprolol (Group B, mean age: 56 +/- 3 yrs, NYHA class II - IV, mean 2.9 +/- 0.2); 8 age-matched healthy volunteers (mean age 49 +/- 3 yrs) served as controls. RESULTS: Compared to controls, in group A isoprenaline-induced lymphocyte cyclic AMP-increase was reduced, CON A evoked IP formation significantly enhanced and isoprenaline-induced inhibition of CON A-evoked IP formation and proliferation almost abolished. In group B, however, all these parameters were not significantly different from controls. CONCLUSION: In CHF patients lymphocyte cyclic AMP response to beta-adrenoceptor stimulation is blunted and the inhibitory effect of cyclic AMP on lymphocyte activation is almost abolished; this could result in a non-regulated increased production and release of proinflammatory cytokines that might contribute to the progression of the disease. Chronic treatment of CHF patients with the beta1 blocker metoprolol (at least partly) restores lymphocyte cyclic AMP responses to beta-adrenoceptor stimulation and the inhibitory effects of cyclic AMP on lymphocyte activation; the resulting "normalization" of the immune function might contribute to the beneficial effects of beta1-blockers in treatment of CHF. PMID- 11403424 TI - Seasonal variation in cotyledoside concentration of Tylecodon wallichii (Harv.) Tolken subsp. wallichii sampled in a krimpsiekte-prevalent region. AB - Krimpsiekte, an economically important neuromuscular affliction of small stock, follows upon ingestion of certain members of the Crassulaceae (plakkies) containing cumulative neurotoxic bufadienolides. Tylecodon wallichii (Harv.) Tolken subsp. wallichii is probably the most important species of the group of plants causing krimpsiekte. The growing tip of the stem and various other plant parts of T. wallichii, when available, were collected monthly. The seasonal variation in cotyledoside content of the plant was measured. Cotyledoside concentration was determined by high performance liquid chromatographic electrospray mass spectrometry analysis (HPLC-ESMS). The cotyledoside concentration in the plant stems fluctuated substantially during the year, but tended to be higher in the cold winter months and increased again in the spring and early summer. Elevated plant stem concentrations corresponded with natural field outbreaks of krimpsiekte, which usually occur during the winter to early summer. The highest cotyledoside concentrations were detected in the flowering stalk. Cotyledoside was not the only component of this type in the plant, as mass spectrometry revealed the presence of other, possibly related, compounds. PMID- 11403425 TI - Studies on the critical water mass, rehydration capability and potential, acute chill tolerance and supercooling point of Argas (Persicargas) walkerae (Acari: Argasidae). AB - The critical water mass, defined as the water mass remaining in a dehydrated tick in the non-ambulatory state, differed only slightly between light and heavy mass groups of Argas walkerae and averaged 23.6% and 23.2%, respectively, in males and 28.4% and 28.0%, respectively, in females. All ticks survived dehydration to 50%, 75% or 100% of their critical water mass, and 95% of them rehydrated during their subsequent incubation at 95% relative humidity (RH) and 28 degrees C for 14 days and regained their ambulatory status. Unfed adults were able to balance water loss frequently over a period of several months. When ticks were repeatedly dehydrated at 0% RH for 14 days, females and males suffered 50% mortality after 16 and 19 cycles of de- and rehydration, respectively, over a period of 278 days and 337 days, respectively. Water itself was not attractive to either dehydrated or non-dehydrated ticks and drinking was not observed. After submergence in water for 3 days, most of the dehydrated adult ticks gained mass. Judged by 50% mortality, larvae tolerated short-term extreme chilling to -24 degrees C, nymphs 1 to -22 degrees C, nymphs II to -20 degrees C, females and males to -19 degrees C. None survived tissue freezing. At a chilling rate of 0.3 degrees C/min, mean supercooling points (SCP) ranged from -25.9 degrees C in eggs to -16.5 degrees C in unfed females. The SCP of all other stages was significantly higher than that of eggs. Mean SCPs of unfed adult ticks dehydrated to 50% or 75% of their critical water mass were significantly lower than that of fully hydrated ticks. The SCPs of ticks acclimated by several weeks exposure to 0 degrees C or to 38 degrees C were significantly lower than those of adult ticks kept constantly at 28 degrees C. PMID- 11403426 TI - Transmission studies on Trichinella species isolated from Crocodylus niloticus and efficacy of fenbendazole and levamisole against muscle L1 stages in Balb C mice. AB - Forty-four Balb C mice, aged 18 weeks were infected with crocodile (Crocodylus niloticus)-derived Trichinella species. Of the infected mice, 32 were randomly divided into two groups each containing equal numbers of males and females; levamisole treated group and fenbendazole treated group. Each group was randomly subdivided into two subgroups as follows: levamisole group (subgroup 1: treated with levamisole on day 35 post infection, and subgroup 2: treated with levamisole on days 35 and 42 post infection) and fenbendazole group (subgroup 1: treated with fenbendazole on day 35 post infection and subgroup 2: treated with fenbendazole on days 35 and 42 post infection). The first subgroups treated on day 35 post infection were slaughtered on day 42 post infection and the second subgroups were treated on day 35 and day 42 post infection and slaughtered on day 49 post infection. Two female mice were infected a day after mating and were slaughtered together with the offspring on day 64 post-infection. Ten infected control mice were given 1 ml distilled water orally as placebo, and five of these were slaughtered on day 42 post infection. The results showed that the mean reproductive capacity index of this strain (RCI) in Balb C mice was 110. There was a significant reduction (P < 0.01) in larval counts in the single treatment groups (day 35) and in the double treatment groups (days 35 and 42) for both anthelmintics when compared the number of parasites in the control groups. After a single treatment, levamisole reduced the infection by 79.9% and fenbendazole by 76.7%. Following double treatments, levamisole reduced the infection by 95.5% and fenbendazole by 99.1%. There was evidence that the infected pregnant mice transmitted the parasite to their offspring. It is not certain whether the parasite was transmitted congenitally or transmammary Alternative ways of controlling the parasite in crocodile farms in Zimbabwe are discussed. PMID- 11403427 TI - Parasites of domestic and wild animals in South Africa. XXXIX. Helminth and arthropod parasites of Angora goats in the southern Karoo. AB - Parasites were collected from 160 Angora goats and kids on the Jansenville Experimental Farm, Eastern Cape Province. Six nematodes were identified to species level and three to generic level. Nematodirus spathiger was the most numerous of the economically important nematodes recovered. It was always present, and fourth stage larvae were collected from untreated goats from August to March, while animals used as tracers also picked up most infection from August to March. Kids born on the farm during October acquired their first nematode infections between 2 and 3 months of age and the intensity of infection increased erratically thereafter to reach a plateau once the kids were 14 months of age. Eight of the nine kids between 3 and 5 months of age and examined between January and March were infected with Moniezia expansa. Five ixodid tick species were collected from the goats of which Rhipicephalus glabroscutatum was the most numerous and prevalent. Its immature stages were present mainly from March to September and adults from July to January. The goats also harboured the biting louse Damalinia limbata and the sucking louse Linognathus africanus. The greatest intensity of infestation with L. africanus occurred on the kids during the first few months of their lives. The larvae of the nasal bot fly, Oestrus ovis were present in kids at 1 month of age, and infestation took place mainly from spring to late summer. PMID- 11403428 TI - A chemically defined medium for the growth of Cowdria ruminantium. AB - Chemically defined media, termed SFMC-23 and SFMC-36, were devised for the in vitro culture of Cowdria ruminantium, the causative agent of heartwater in domestic ruminants. Both media were based on Dulbecco's modified Eagle's medium nutrient mixture Ham F-12 (DME/F-12) containing various supplements. Medium SFMC 23 and SFMC-36 supported the long-term growth of the Welgevonden stock of C. ruminantium for a total of 55 and 28 passages, respectively, with regular passage intervals of 3 days. Using SFMC-23, split ratios varied from 5-10, depending on which host cell line was used. Other stocks of C. ruminantium (Sankat, Blaauwkrantz, Senegal) were successfully propagated for a test period of ten passages. PMID- 11403429 TI - Tick infestation of Borana cattle in the Borana Province of Ethiopia. AB - A study was conducted to identify and determine burdens of ticks infesting Borana cattle in the Borana Province of Ethiopia. Rhipicephalus pulchellus, Rhipicephalus pravus, Rhipicephalus muhsamae, Rhipicephalus praetextatus, Rhipicephalus evertsi evertsi, Amblyomma gemma, Amblyomma variegatum, Amblyomma cohaerens, Amblyomma lepidum, Hyalomma truncatum, Hyalomma marginatum rufipes and Boophilus decoloratus were identified on the cattle. Their burdens ranged from 658-1,554 with a mean of 1,205 ticks per animal. Out of the total 6,025 ticks collected, about 82% were R. pulchellus, followed by R. pravus (about 8%) and A. gemma (5%). The other nine species of ticks accounted for only 5% of the total burdens. It is suggested that a strategic tick control method, aimed mainly at reducing the R. pulchellus burdens, with the objective of allowing a reasonable number of ticks to remain on cattle for the maintenance of endemic stability to tick-borne diseases be instituted. PMID- 11403430 TI - Closure of the ductus arteriosus of indigenous South African goats at high altitude. AB - The closure of the ductus arteriosus (DA) of 31 indigenous South African goats, whose ages ranged from 30 days prenatal to 60 days postnatal, were studied at an altitude of 1,514 m above sea level by vascular injection as well as histologically and ultrastructurally. The vascular injection results showed that functional occlusion started from the pulmonary end of the DA in kids 6 days old and progressed to the aortic end in kids 8 days old. Histologically, anatomical obliteration was observed in kids from 35 days of age. The functional closure was preceded by enlargement of the subendothelial region, progressive intimal thickening, presence of subendothelial vacuolization and endothelial detachment. There was radial orientation of the subintimal smooth muscle cells and subsequent migration towards the intima. The inner tunica media contained mast cells and areas of cytolysis. Following functional closure, the subendothelial region showed migrating subintimal smooth muscle cells with extensive cytoplasmic processes and, ultrastructurally a fragmented internal elastic lamina. In 15-day old kids there were prominent, progressively enlarged cisternae of the rough endoplasmic reticulum and numerous free, dispersed ribosomes. In kids 19 and 25 days old, there was, additionally, rarefaction of the cell cytoplasm and appearance of intracellular myofibrils and extra cellular collagen in the surrounding amorphous matrix, which culminated in the complete anatomical closure of the DA in 35-day-old kids. PMID- 11403431 TI - Refugia--overlooked as perhaps the most potent factor concerning the development of anthelmintic resistance. AB - Anthelmintic resistance involving particularly the gastrointestinal nematodes of small ruminants is escalating globally, to the extent that in certain countries, such as South Africa, it has already reached alarming proportions, and is affecting practically all the anthelmintics. In this paper it is argued that the high levels of resistance in nematodes of veterinary importance indicate that the drugs have been used incorrectly. It is suggested that the phenomenon of refugia plays a much more important role in the selection of anthelmintic resistance than other phenomena that are more frequently investigated and recommended for counteracting it, such as reduced drenching frequency and avoiding under-dosing. While refugia is commonly mentioned in passing in most papers on anthelmintic resistance, it is, almost without exception, not incorporated in the final control/ management proposals. On the strength of the conclusions arrived at in the present paper strategies such as the drench-and-move system in which all the animals in a flock are drenched before they are moved to pastures containing few or no worms in refugia, and the system of strategic drenching on safe pastures should be condemned and never recommended. If such strategies are indeed unavoidable, the farmer should be warned that the farming system would probably not be sustainable even in the short term, in view of the generally high levels of resistance already present in most of the important sheep-producing regions. Farmers should be educated to consider refugia above all else when designing worm management programmes. Finally there seems to be too much complacency concerning the possibility that anthelmintic resistance may also escalate in cattle, eventually to reach the proportions that it has in sheep. PMID- 11403432 TI - Antibodies to Newcastle disease virus in the sera of indigenous chickens in Oodi, Kgatleng, Botswana. AB - A serological survey was conducted to determine the prevalence of antibodies to Newcastle disease virus in apparently healthy and unvaccinated adult indigenous chickens. Haemagglutination inhibiting antibodies to Newcastle disease virus were found in the sera of 51 out of 89 (57.3%) chickens sampled. PMID- 11403433 TI - The establishment, composition and severity of infection of gastro-intestinal parasites and their impact on productivity of Tswana kids in southern Botswana. AB - The presence of gastro-intestinal parasites in Tswana kids (n = 7) aged 1-3 weeks was studied for a period of 6 months at the Botswana College of Agriculture. The aims of this study were to find the time when they first contracted internal parasite infections, as well as to determine the severity of the infections and also its relation to production indicators (body mass and packed cell volume) of the kids as they grew older. The results indicate that they contracted coccidial and roundworm infections at approximately one month of age or immediately thereafter. The most prevalent internal parasite was coccidia, which occurred throughout the study period followed by roundworms and the least was the tapeworm, Moniezia expansa. Generally, the infection levels of all internal parasites were lower than the critical mean log (faecal oocyst/egg count + 1) of 3.3 inferred to cause reduced production in small stock. The correlation coefficients were all positive; 0.4-0.9 for individual internal parasites and production indicators, indicating that these internal parasites did not have any adverse effects on production. It was concluded that there was no need to treat kids of this age group for internal parasites. PMID- 11403434 TI - Hosts of the immature stages of the rhinoceros tick, Dermacentor rhinocerinus (Acari, Ixodidae). AB - The collection of the immature stages of Dermacentor rhinocerinus from host animals is described for the first time. These are the bushveld gerbil (Tatera leucogaster, from which larvae and nymphs were collected, the red veld rat (Aethomys crysophilus), from which a larva was collected, and the Natal multimammate rat (Mastomys natalensis), from which a nymph was collected. The rodents were examined in a nature reserve in which there are white rhinoceroses (Ceratotherium simum). PMID- 11403435 TI - Oral administration of mebendazole failed to reduce nematode egg shedding in captive African gazelles. AB - Idiosyncracies are observed in captive wild animals as regards the pharmacokinetics and efficacy of anthelmintics. This could be attributed to such factors as differences in host's metabolism, irregular distribution of anthelmintics due to the way they are administered and worm resistance to anthelmintics. Previously mebendazole was found to be poorly effective when administered in feed. An experiment was conducted to evaluate the efficacy of mebendazole when administered at the dosage rate of 15-20 mg/kg body weight to gastrointestinal nematodes in captive gazelles. Fifty-eight adult gazelles (Gazella cuvieri) were divided into four groups: T1 (animals dosed orally, directly into the mouth), T2 (treated orally, mixed in the water of a herd), T3 (treated orally, mixed in the water of one animal), and T4 (not treated). Individual faecal samples were taken before treatment, and 15 days thereafter. Mean percentages of reduction of egg shedding were calculated for Nematodirus spp., other trichostrongyles, total trichostrongylids, Trichuris spp. and total nematodes. No statistically significant differences were detected between the treatment groups and the control group or among the animals in the three treatment groups. PMID- 11403436 TI - Study of retention in micellar liquid chromatography on a C8 column by the use of linear solvation energy relationships. AB - Linear solvation energy relationships (LSERs) are used to investigate the fundamental chemical interactions governing the micellar liquid chromatographic retention of 22 aromatic compounds (11 benzene derivatives and 11 aromatic polycyclic hydrocarbons) in 80 mobile phases on a C8 column. The systems studied involve combinations of 0.050 to 0.140 M sodium dodecyl sulfate or cetyltrimethylammonium bromide, with 0 to 10% methanol, n-propanol, and n-butanol as mobile phase modifiers. The ability of the LSERs to account for the chemical interactions underlying solute retention is shown. A comparison of predicted and experimental retention factors suggests that LSER formalism is able to reproduce adequately the experimental retention factors of the solutes studied in the different experimental conditions investigated. PMID- 11403437 TI - Elution parameters in constant-pressure, single-ramp temperature-programmed gas chromatography. AB - The dependence of the degree of interaction of a solute with the stationary phase at the time of its elution from the column in temperature-programmed GC is best described by interaction level of the solute. The latter represents the fraction of a solute residing in the stationary phase relative to the total amount of the solute. A simple approach to the evaluation of interaction levels of eluting solutes in a single-ramp temperature program is proposed. In a single-ramp temperature program having no preceding temperature plateau, all solutes that elute at temperatures that are about 60 degrees C higher than the initial temperature of the heating ramp elute with nearly the same interaction levels that can be found as exp(-r), where r is dimensionless heating rate. A specially designed temperature plateau preceding the ramp causes all solutes eluting during the entire time of the ramp to elute with nearly the same interaction levels equal to exp(-r). A transformation of the interaction level of a solute into its retention factor or mobility factor (a fraction of a solute in a mobile phase in relation to the total amount of the solute) and vice versa is also described. PMID- 11403438 TI - Influence of solvents and gas chromatographic injector conditions on the detectability of nitroaromatic compounds. AB - We investigated the influence of four common solvents and of several liner packings of a split/splitless injector on the gas chromatographic behavior of trinitrotoluenes and related nitroaromatic compounds. The highest peaks are observed using toluene in combination with an empty liner or with a prepacked CarboFrit liner. In particular, the peaks of trinitrotoluene isomers and 1,3,5 trinitrobenzene significantly decreased or even totally disappeared when using quartz wool or glass wool, even when treated with dimethylchlorosilane. Similiar peak reductions are obtained with methanol or acetonitrile. Effects of decreasing peak are accompanied by the formation of two additional products when using methanol. PMID- 11403439 TI - Study of Croatian non-aromatic naphthalane constituents with skeletons analogous to bioactive compounds. AB - Non-aromatic Croatian naphthalane (NAN), shown to be efficace in the treatment of psoriasis vulgaris, was studied in order to improve our understanding of its constituents, which may be potentially responsible for its bioactivity. The components steranes and hopanes were analysed. Since NAN is a complex mixture of hydrocarbons, high-resolution GC and GC-MS were applied as the methods of choice in the study. The GC chromatogram of NAN showed a remarkable cluster in which sterane peaks prevail and composed 33+/- 1% of the sample. Identified steranes (by GC-MS) represented almost half of the cluster (48%). They were in the range from norcholestanes up to propyl cholestanes. The amount of alpha-steranes (8.9%) was higher than of beta-steranes (6.4%) and regular steranes (15.3%) dominated in the ratio 17:1 over the rearranged ones (0.9%). Steranes conserved the skeleton of bio-precursors and remained analogues of bioactive compounds, such as of vitamins D and some hormones and corticosteroids. Pentacyclic hydrocarbons hopanes, as derivatives of bacteriohopantetraol which is the physiological equivalent of cholesterol, made 4.7+/-0.2% of NAN. PMID- 11403440 TI - Determination and comparison of competitive isotherms by rectangular pulse method and frontal velocity analysis method. AB - Two different approaches, the rectangular pulse method and a new method, the frontal velocity analysis method are applied to the determination of competitive isotherms for two systems on ODS-silica. Both sets of experimental data are found to fit well to the competitive-Langmuir isotherm equation. The data obtained from the rectangular pulse method and the frontal velocity analysis method are in general agreement with each other and the best coefficients of the competitive isotherms obtained with the two methods are close. This shows that the simpler and easier method, the frontal velocity analysis method can be used to determine binary competitive isotherms. PMID- 11403441 TI - Gas chromatographic study of the volatile products from co-pyrolysis of coal and polyethylene wastes. AB - The aim of this study was to determine the volatile products distribution of co processing of coal with two plastic wastes, low-density polyethylene from agriculture greenhouses and high-density polyethylene from domestic uses, in order to explain the observed decrease in coal fluidity caused by polyethylene waste addition. Polymeric materials, although they are not volatile themselves, may be analysed by gas chromatography through the use of pyrolysis experiments. In this way, a series of pyrolysis tests were performed at 400 and 500 degrees C in a Gray-King oven with each of the two plastic wastes, one high-volatile bituminous coal and blends made up of coal and plastic waste (9:1, w/w, ratio). The pyrolysis temperatures, 400 and 500 degrees C, were selected on the basis of the beginning and the end of the coal plastic stage. The organic products evolved from the oven were collected, dissolved in pyridine and analysed by capillary gas chromatography using a flame ionization detector. The analysis of the primary tars indicated that the amount of n-alkanes is always higher than that of n alkenes and the formation of the alkenes is favoured by increasing the pyrolysis temperature. However, this effect may be influenced by the size of the hydrocarbon. Thus, the fraction C17-C31 showed a higher increase of n-alkenes/n alkanes ratio than other fractions. On the other hand, the difference between the experimental and estimated values from tars produced from single components was positive for n-alkanes and n-alkenes, indicating that co-pyrolysis of the two materials enhanced the chemical reactivity during pyrolysis and produced a higher conversion than that from individual components. PMID- 11403442 TI - Selectivity of solid-phase extraction phases in the determination of biodegradation products. AB - The extraction techniques connected with gas chromatography were used to describe quantitatively and qualitatively the biodegradation process. We investigated the biodegradation of hydrocarbons and non-ionic surfactants. Solid-phase extraction (SPE) and liquid-liquid extraction were used for the isolation of the non degraded compounds and their degradation products. The selectivity of SPE has a significant influence on the isolation and preconcentration of organic compounds from water. PMID- 11403443 TI - Wet effluent diffusion denuder technique and determination of volatile organic compounds in air. I. Oxo compounds (alcohols and ketones). AB - The wet effluent diffusion denuder (WEDD) preconcentration technique for the determination of methanol, ethanol, 1-propanol, 2-propanol, 1-butanol, 1 pentanol, acetone, methyl ethyl ketone, diethyl ketone and methyl n-propyl ketone in air is discussed. The compounds are continuously collected into a thin film of deionized water flowing down the inner wall of the cylindrical wet effluent diffusion denuder. The concentrate is analysed by gas chromatography. Detection limits of alcohols and ketones are as low as 1 microg/l (GC-flame ionization detection) and/or 1 ng/l (GC-MS). This technique could be applicable for the continuous monitoring of ppb (v/v) levels of both alcohols and ketones in the air. PMID- 11403444 TI - Solid-phase microextraction of hop volatiles. Potential use for determination and verification of hop varieties. AB - The composition of hop essential oil is an important tool for evaluation of hop quality. As each hop variety has a typical essential oil pattern (fingerprint), hop oil analyses can be used to distinguish between hop varieties. The headspace solid-phase microextraction (SPME) method as described in this contribution is a simple sample preparation technique and represents an alternative procedure for essential oil fingerprint determination. Different SPME parameters (extraction temperature, extraction time and sample mass) were studied and the results were compared with those obtained by the routine distillation method. It is shown that SPME results can be used for determination and verification of varieties grown in Slovenia by means of principal components analysis. PMID- 11403445 TI - Solid-phase microextraction method for gas chromatography with mass spectrometric and pulsed flame photometric detection: studies of organoarsenical speciation. AB - The development, optimization, and application of a novel method for arsenic speciation based on capillary gas-liquid chromatography with simultaneous quadrupole ion-trap mass spectrometric (MS) detection and pulsed flame photometric detection (PFPD) is described. The method couples the sensitive arsenic-selectivity of PFPD with the structure elucidation capability of molecular MS detection for the determination of trace levels of unknown organoarsenicals in complex matrices. The conditions that affect the PFPD response in the presence of interfering species were optimized using the sequential Simplex algorithm for three key factors: gate delay (18.3 ms), gate width (9.1 ms), and combustion gas composition (16.6 ml/min H2). Complete discrimination in the PFPD of the arsenic signal from interfering S-, C-, and OH emitting species that are problematic in existing methods was achieved. Additionally, a revised interpretation of our previously reported mechanism [J. Chromatogr. A 807 (1998) 253] for the dithiol derivatization and subsequent GC-MS determination of dimethylarsinic acid is presented. PMID- 11403446 TI - Determination of organochlorine pesticides and their metabolites in soil samples using headspace solid-phase microextraction. AB - An analytical procedure was developed using headspace solid-phase microextraction (HS-SPME) for the determination of organochlorine pesticides (OCPs) and their metabolites in sandy soil samples. The developed procedures involving fiber selection, temperature effect, absorption time, soil matrix and the addition of solvents of different polarity were optimized. Also, the results were compared to those achieved using Soxhlet extraction standard method. The 100-microm polydimethylsiloxane (PDMS) and 65-microm PDMS-divinylbenzene showed good extraction efficiency for 18 organochlorine pesticides. An increase in the extraction efficiency of organochlorine pesticides and the metabolites was observed when the temperature increased, and an optimum temperature of 70 degrees C for extracting OCPs was obtained. The application of other hydrophilic solvents had different effects on the extraction of organochlorine pesticides and the metabolites. Higher responses of OCPs were obtained when 5 ml of water was added to the soil. Good linearity of OCPs between 0.2 and 4 ng/g soil was observed. The relative standard deviation was found to be lower than 25%. Also the limits of detection were between 0.06 and 0.65 ng/g, which were lower than those obtained using Soxhlet extraction. Moreover, the optimized HS-SPME procedure was applied to the analysis of OCPs in certified reference material (CRM) 804-050 soil and compared with Soxhlet extraction procedure. Results obtained in this study were in good agreement with those obtained using Soxhlet extraction. The mean values obtained using HS-SPME technique were in the range of 16.5 to 1459.6 mg/kg, which corresponds to the recoveries of 68% to 127% of the certified values of CRM soil. PMID- 11403447 TI - Analysis of Origanum vulgare volatiles by direct thermal desorption coupled to gas chromatography-mass spectrometry. AB - Volatile components of samples of a population of Spanish Origanum vulgare have been analyzed by direct thermal desorption coupled to GC-MS. The method is fast and reliable and requires a low amount of sample, allowing analysis of leaves and flowers from a single individual plant. Volatile yield is highly variable among individual plants and concentration also presents a high variation for most Origanum volatile compounds, linalool being the main component in most samples. Statistical analyses are applied in order to find patterns in composition data. PMID- 11403448 TI - Analytical aspects of carrier ampholyte-free isoelectric focusing. AB - The applicability of carrier ampholyte-free isoelectric focusing (CAF-IEF) for analyses of ampholytes is demonstrated. The suggested method is based on the principle of both side regulated ionic matrix in CAF-IEF. A sharp step of pH is created in the column filled with a sample dissolved in a background electrolyte by influence of current and solvolytic fluxes. Here, ampholytes are focused upon. The magnitude of the step, its velocity and direction of its movement can be regulated electrically. In this manner, favorable separation properties of the system can be set up, even during the run. This brings several advantages over conventional methods. The principles of the separation can be easily changed, permitting selective pre-concentration (trapping) of minor components by processing large amounts of a sample to be preformed, effective isotachophoresis or IEF pre-separation and final electrophoretic analysis in one run. Advantages of these combinations are discussed together with the right choice of the working electrolyte. A 1000-fold increase in amount of substance in a column can be achieved for both isotachophoresis and capillary zone electrophoresis combined with CAF-IEF pre-concentration at reasonable working conditions. It enables a limit of detection at the nmol/l level with a concentration factor of about 10(7) to be reached. PMID- 11403449 TI - Determination of sulfametoxazole, sulfadiazine and associated compounds in pharmaceutical preparations by capillary zone electrophoresis. AB - A capillary zone electrophoresis method is presented to separate sulfadiazine, sulfamethoxazole, trimethoprim, bromhexine and guaiacol by using a fused-silica capillary (60.2 cm x 75 microm I.D.). The separation was carried out at 30 kV and 25 degrees C in a 15 mM phosphate buffer adjusted to pH 6.2 as electrolyte. Under these conditions, the run time was 6 min and the limits of quantification were about 1 mg/l for every component. The method was applied to pharmaceutical preparations and the results provided recoveries close to 100%. PMID- 11403450 TI - Capillary electrochromatography of tricyclic antidepressants on strong cation exchangers with different pore sizes. AB - Four cation-exchange materials, possessing propanesulfonic acid ligands, for use in capillary electrochromatography were prepared from different commercially available 5-microm bare-silica particles ranging from 80 to 800 A in pore size. The performance of the materials was investigated at different compositions of the mobile phase (pH, ionic strength, and acetonitrile content) using tricyclic antidepressants and related quaternary ammonium analogues as test analytes. The wide-pore materials promoted pore flow, but this had no positive influence on the performance. The small-pore (highest surface area) particles gave, as could be expected, the best selectivity. PMID- 11403451 TI - Separation of polyethylene glycol oligomers using inverse temperature programming in packed capillary liquid chromatography. AB - Inverse temperature programming in packed capillary liquid chromatography coupled to evaporative light-scattering detection has been used to resolve native polyethylene glycol (PEG) oligomers. The model compound, PEG 1000, was separated on a 300 mm x 0.32 mm I.D. capillary column packed with 3 microm Hypersil ODS particles with acetonitrile-water (30:70, v/v) as mobile phase. The retention of the PEG oligomers increased with increasing temperature, different from what is commonly observed in liquid chromatography. The retention times of the oligomers were approximately doubled for each 25 degrees C increment of the column temperature in the temperature range 30-80 degrees C. The oligomers were almost unretained and co-eluted at a column temperature of 30 degrees C. At 80 degrees C a baseline separation of more than 22 peaks was obtained, but the last eluting peaks were severely broadened and all oligomers did not elute. When a negatively sloped temperature ramp from 80 to 25 degrees C at -1.5 degrees C/min was applied, the peak shapes were improved, additional peaks were detected and the analysis time was reduced by 48%. In the temperature programming mode, the intra day precision of the retention times ranged from 0.5 to 5.8% (n=5). PMID- 11403452 TI - Chlorotrimethylsilane as a reagent for gas chromatographic analysis of fats and oils. AB - Chlorotrimethylsilane can be used as a reagent to transform triglycerides into volatile fatty esters. The volatile esters can then be analysed by GC. The results are fully comparable to those obtained by alternative methods used worldwide. The new one-step method can transesterify acylglycerides and esterify free fatty acids at the same time. Chlorotrimethylsilane is cheaper than BF3-MeOH and is likely to permit the use of different alcohols. PMID- 11403453 TI - Liquid-core waveguide technology for coupling column liquid chromatography and Raman spectroscopy. AB - The on-line coupling of liquid chromatography (LC) and Raman spectroscopy (RS) via an entirely plastic liquid-core waveguide (LCW) was optimized in terms of excitation wavelength of the laser, especially in relation to the fluorescence background, and the length of the LCW. Excitation at 632.8 nm (He-Ne laser) was found to be a good compromise between a wavelength long enough to strongly reduce the fluorescence background and, on the other hand, short enough to avoid (re) absorption of laser light and Raman signals by H2O in LCWs of considerable length. This conclusion is supported by a theoretical discussion on the optimization of LCW lengths as function of the excitation wavelength for H2O and 2H2O. When using the He-Ne laser the optimum length is approximately 50 cm for H2O; this corresponds to a detection cell volume of 19 microl for an LCW of 220 microm I.D., which is fully compatible with conventional-size LC. The influence of an organic modifier, usually necessary for reversed-phase LC, on the free spectral window was evaluated. The potential applicability of LC-LCW-RS was shown for a mixture of adenosine 5'-monophosphate (AMP), guanosine 5'-monophosphate (GMP) and uridine 5'-monophosphate (UMP), utilizing an aqueous eluent without the addition of a modifier. Improved detectability was achieved by using the stopped flow mode and applying a large-volume-injection procedure (injection volume: 200 microl). Under these conditions, the limit of identification for AMP, GMP and UMP was in the 0.1-0.5-mg/ml range. PMID- 11403454 TI - Contribution of axial dispersion to band spreading in perfusion chromatography. AB - A new interpretation of the band spreading data in perfusion chromatography is proposed by investigating the relative importance of axial dispersion in perfusive beds. Elution chromatography of proteins (bovine serum albumin and lysozyme) under non-retained conditions on two kinds of reversed-phase perfusive supports (POROS R1/H and POROS R2/H), which have different pore structures, were carried out to obtain the axial dispersion data. The Knox equation and some empirical correlations for dispersion coefficients in porous media were applied to correlate the experimental data. The influences of particle properties, solute molecular sizes and flow velocity on the dispersion coefficient were elucidated. Axial dispersion was recognised to be the main contributor to peak broadening in perfusion chromatography. The dependence of the height equivalent to a theoretical plate on flow-rate was found to be the result of the velocity dependence of the axial dispersion. The dispersion coefficient in a perfusive column can be well represented both by a power-law relationship and a correlation derived based on stochastic theory. Pursuant to these, it was found that pore size distribution of the perfusive particles and solute molecular size are important parameters, which influenced the dispersion results significantly. PMID- 11403455 TI - Fractionation of low-molecular-mass heparin by centrifugal partition chromatography in the ion-exchange displacement mode. AB - Centrifugal partition chromatography in the ion-exchange displacement mode allowed a preparative and efficient fractionation of low-molecular-mass heparins from enoxaparin sodium. Amberlite LA2--a lipophilic liquid secondary amine--was chosen as a weak anion exchanger. The biphasic system methyl isobutyl ketone water was selected. Protonated LA2 (10%, v/v) was added to the organic stationary phase. Hydroxide (Na+, OH-) was chosen as a displacer in the aqueous mobile phase. The observed pH and concentration profiles are typical of displacement chromatography, as supported by numerical simulation. The Dubois test for the analysis of sugar content and an analysis of sulfur content (and consequently sulfatation rate) were carried out to monitor the effectiveness of the procedure. Moreover, the fractions were analyzed by high-performance size-exclusion chromatography and the 1H NMR spectra confirmed the fractionation of the sample of enoxaparin sodium. PMID- 11403456 TI - Comparison of various liquid chromatographic methods for the analysis of avermectin residues in citrus fruits. AB - Various liquid chromatographic (LC) techniques for analyzing avermectin (Abamectin) were compared after extraction of residues from citrus fruit samples by matrix solid-phase dispersion (MSPD). LC with UV and fluorescence detection were used as also was LC coupled to the mass spectrometer by an electrospray interface. The results obtained by the three methods were compared in terms of sensitivity and selectivity. The combination of MSPD extraction and LC with fluorescence detection have made it possible to quantify 0.5 microg kg(-1) of Abamectin in 0.5 g of orange sample, with an overall average recovery of 94%. The procedure provides a simple and sensitive method for monitoring Abamectin residues in citrus fruit at the levels required by legislation. PMID- 11403457 TI - Determination of polar organophosphorus pesticides in aqueous samples by direct injection using liquid chromatography-tandem mass spectrometry. AB - It was demonstrated that four out of six of the very polar organophosphorus pesticides (OPs), i.e. acephate, methamidophos, monocrotophos, omethoate, oxydemeton-methyl and vamidothion, could not be extracted from water using commonly available SPE cartridges. In addition, GC analysis on all six compounds was found to be troublesome due to their polar and thermolabile character. This initiated the development of an alternative highly sensitive and selective method for the determination of the above mentioned very polar OPs in water, based on LC MS. Large volume (1 ml) water samples were directly injected onto an RP18 HPLC column with a polar endcapping. The latter was essential for obtaining retention and maintaining column performance under 100% aqueous conditions during the sampling. The compounds were ionized using atmospheric pressure chemical ionization and detected on a tandem mass spectrometer operated in multiple reaction-monitoring mode. The detection limits were in the range of 0.01-0.03 microg/l. Compared to conventional GC methods, the developed LC-MS procedure is very straightforward, fast and more reliable. This application demonstrates the applicability of LC-MS for analysis of polar OPs in surface, ground and drinking water, as a more favourable alternative to GC. PMID- 11403458 TI - Determination of benzoylureas in tomato by high-performance liquid chromatography using continuous on-line post-elution photoirradiation with fluorescence detection. AB - A photochemically induced fluorescence post-column method, with HPLC separation and fluorescence detection, was developed for the determination of five naturally non-fluorescent benzoylurea insecticides: diflubenzuron, triflumuron, hexaflumuron, lufenuron and flufenoxuron. The applicability of the method to the determination of insecticides in spiked tomato was evaluated. Samples were extracted into ethyl acetate and further cleaned-up by solid-phase extraction using an aminopropyl-bonded silica cartridge. The interferences due to the matrix effect were eliminated using matrix matched standards. Linear dynamic ranges were established over more than two orders of magnitude. The limits of detection ranged from 5 to 21 ng ml(-1) (or 0.5 and 2.1 microg kg(-1) in the vegetable samples), with relative standard deviations lower than 5.0%, using blank tomato extract. Mean recoveries ranged from 79 to 102%. PMID- 11403459 TI - Speciation of arsenic and selenium compounds by ion-pair reversed-phase chromatography with electrothermic atomic absorption spectrometry. Application of experimental design for chromatographic optimisation. AB - An off-line system is proposed consisting of ion-pair reversed-phase liquid chromatography, collections of fractions at the outflow of the column and furnace atomic absorption spectrometry. The so-called system allowed determination of both arsenic and selenium species mainly found in the environment and in mammals (arsenite, arsenate, monomethylarsonate, dimethylarsinate, selenite, selenate, selenocystamine, selenocystine, selenomethionine and selenoethionine). In order to study the retention behaviour of these compounds and to estimate the optimal conditions for the chromatographic separation, central composite designs were used to evaluate the influence of the eluent parameters such as pH, tetrabutylammonium phosphate (TBA) concentration and sodium hydrogenphosphate amounts. The retention factors of each species and the selectivity were established as response criteria. Response surfaces and isoresponse curves were drawn from the mathematical models and enabled one to determine the optimal conditions and to visualise the method robustness. The predicted optimal zone was situated at pH 5.5-6.5, 4.0 mM Na2HPO4 and 3.0-4.0 mM TBA. Regression models suggested linearity for the studied compounds in the range 25-200 microg selenium and arsenic per litre investigated. PMID- 11403460 TI - Critical study of and improvements in chromatographic methods for the analysis of type B trichothecenes. AB - Various analytical methods used in the analysis of type B trichothecenes (deoxynivalenol, nivalenol, 3- and 15-acetyldeoxynivalenol) in cereals were compared and optimised in this work. These methods use either GC-electron-capture detection (ECD) of trimethylsilyl, trifluoroacetyl and heptafluorobutyryl derivatives or HPLC with UV or photodiode array detection of analytes. A new HPLC procedure using fluorescence detection prior derivatisation with coumarin-3 carbonyl chloride has been also tested. Five extraction solvents and two solid phase extraction cartridges (silica, Florisil) plus a especial clean-up column (MycoSep 225) were compared in order to obtain the best recovery of the mycotoxins with minimal presence of coextractives in the chromatograms. The chosen extraction solvent was a mixture of acetonitrile-water (84:16, v/v). The MycoSep 225 column was chosen as the best alternative for clean-up of grain samples. For GC-ECD analysis, derivatisation of analytes with heptafluorobutyric anhydride prior the final determination was chosen as the most suitable procedure. HPLC-photodiode array (at 221 nm) analysis was more suitable for determination of type B trichothecenes than HPLC of the fluorescent coumarin-3 carbonyl derivatives. Recoveries obtained in spiked corn, rice and wheat are reported. The utility of the proposed methodology was assayed in cereal cultures of various Fusarium strains. PMID- 11403461 TI - Visualization of solute migration in chromatographic columns influence of the frit porosity. AB - The dual-perspective, on-column detection method previously described was used to observe the effects of the inlet frit on the profiles of chromatographic bands. Visualization of bands of iodine was achieved by injecting its dilute solutions in carbon tetrachloride into a glass column packed with a C18-bonded silica and eluted with carbon tetrachloride, which has the same refractive index as the packing material. The bands were photographed on-column with two standard 35-mm SLR cameras oriented at right angle. The photographs were scanned and the digitized images of the sample bands analyzed with proper software. A number of columns, as similar as possible, were fitted with different 2- and 10-microm porosity stainless steel frits. Subsequent analysis of the digitized band images revealed irregularities in the band shape resulting from frit contributions to band dispersion. The 2-microm frits produced more dramatic effects overall than the coarser frits. Local axial dispersion coefficient values, expressed as local reduced plate height, were calculated. The results demonstrate the possibly damaging effects of the frit on the band shape. PMID- 11403462 TI - Determination of heparin on intraocular lens surfaces by ion chromatography. AB - A sensitive and selective method has been developed for the determination of heparin on heparin coated PMMA, poly(methyl methacrylate), intraocular lenses. Heparin was hydrolysed to glucosamine and glucuronic acid, and the content of glucosamine was determined using ion chromatography with pulsed amperometric detection. In order to verify that a complete hydrolysis was obtained for the heparin on the coated intraocular lenses, electron spectroscopy for chemical analysis (ESCA) was used for analysing traces of sulphur on the lens surfaces. The sensitivity of the method allows quantitative determination of 150 ng of heparin on one individual lens. The new method was compared to a standard spectrophotometric method, measuring the colour intensity of a heparin toluidine blue complex. Correlation between the methods was shown for samples prepared from PMMA lenses coated with different amounts of heparin. PMID- 11403463 TI - High-performance anion-exchange chromatography-electrospray mass spectrometry for investigation of the substituent distribution in hydroxypropylated potato amylopectin starch. AB - The use of high-performance anion-exchange chromatography (HPAEC) with pulsed amperometric detection (PAD) coupled on-line with electrospray mass spectrometry (ESI-MS) for analysis of the substitution pattern in chemically modified starch, has been investigated. In order to characterise the distribution of substitution groups along the polymer chain, hydroxypropylated potato amylopectin starch (HPPAP) was subjected to enzymic hydrolysis, followed by analysis of the degradation products by HPAEC-PAD-MS. When using conventional chromatographic techniques for characterisation of enzymic hydrolysates, standard compounds are required for identification of the hydrolysis products. However, the on-line coupling with ESI-MS allowed identification of all products obtained, substituted as well as unsubstituted, and also of those compounds that co-eluted, without the need for standards. Further, HPAEC-PAD-MS was shown to be useful for analysis of the substitution pattern in modified starch; from results obtained it was suggested that the hydroxypropyl groups were homogeneously distributed in the amylopectin molecule. It was also shown that the starch hydrolysing enzymes were hindered by the hydroxypropyl groups and preferentially cleaved glucosidic linkages between unsubstituted glucose units. PMID- 11403464 TI - Effect of the mobile phase on the retention behaviour of optical isomers of carboxylic acids and amino acids in liquid chromatography on bonded Teicoplanin columns. AB - Conditions for separation of enantiomers of underivatized amino acids phenyl glycine and tryptophan and of mandelic acid as test compounds were studied on a Chirobiotic T column packed with amphoteric glycopeptide Teicoplanin covalently bonded to the surface of silica gel. The effects of the mobile phase composition on the retention and selectivity under analytical conditions, on the profile of the adsorption isotherms of the enantiomers and on the overloaded separation were investigated. The concentration of ethanol or of methanol in aqueous-organic mobile phases and the pH of the mobile phase affect not only the retention and selectivity, the saturation capacity and the isotherm profile, but also the solubility of the acids, which should be taken into account in development of preparative separations. A compromise between the separation selectivity and the solubility should be made in selecting the mobile phase suitable to accomplish preparative separations at acceptable production rate and throughput of the operation. PMID- 11403465 TI - Chromatographic purification and characterization of B-phycoerythrin from Porphyridium cruentum. Semipreparative high-performance liquid chromatographic separation and characterization of its subunits. AB - A fast preparative two-step chromatographic method for purification of B phycoerythrin from Porphyridium cruentum is described. This biliprotein was homogeneous as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis yielding three closely migrating bands corresponding to its three subunits. Baseline separation of its alpha-, beta- and gamma-subunits was achieved by a reversed-phase HPLC gradient semipreparative method with a C4 large pore column and a solvent system consisting of 0.05% trifluoroacetic acid (TFA) in water and 0.05% TFA in acetonitrile. B-Phycoerythrin in different aggregation states and its subunits have been spectroscopically characterized. Hexameric B phycoerythrin has similar secondary and tertiary structure than dissociated B phycoerythrin determined by circular dichroism. PMID- 11403466 TI - Validatibility of a capillary isoelectric focusing method for impurity quantitation. AB - A strategy is presented for examining the validatability of a capillary isoelectric focusing (cIEF) method, intended for quantitation of product-related impurities in a protein drug substance, according to guidelines published by the International Conference on Harmonization (ICH). The results of this study demonstrate the suitability of cIEF as an analytical method for the quantitation of two product-related impurities in a protein drug substance: a monodeamidated degradation product and an aggregated form of the parent molecule. A range of impurity levels was generated by spiking the isolated impurity species, into a representative production lot of the drug substance. Six impurity spike levels (0.5-12% impurity for deamidated species and 0.5-8% impurity for aggregated species) were analyzed in triplicate. Measurement of impurity peak area percent in the spiked samples provided the data for computing specificity, accuracy, precision, linearity and limit of quantitation (LOQ) for the impurities. Accuracy, defined as the agreement of peak area percent for impurity species with the theoretical impurity percentage from the spike ratio, was 85-96% for the deamidated species and 73-97% for the aggregated species. A linear relationship was found between the measured area percent and the theoretical percent impurity for both impurity species (coefficient of determination, r2=0.9994 for deamidated species and =0.9827 for aggregated species). Precision (repeatability) studies demonstrated a low relative standard deviation (RSD) value (<6%) at all spike levels for both impurity species. Intermediate precision and reproducibility were evaluated by simulating many of the multivariable testing conditions expected during the life cycle of an analytical method, such as multiple equipment and laboratories. Repeated analyses of the drug substance under these varied conditions, yielded RSD values of <20%, for both impurity species. The LOQ, defined as the lowest impurity level where both accuracy and precision were achieved, was assigned at the 0.5% impurity level for both impurity species. This work illustrates a successful strategy in applying the ICH validation guidelines for impurity analytical methods to a cIEF method. Moreover, the data demonstrate the ability of cIEF to be used reliably as an analytical method for impurity quantitation. PMID- 11403467 TI - Liquid chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry determination of N-methylpyrrolidinone in riverine biofilms. AB - A rugged procedure utilizing reversed-phase liquid chromatography with positive ion electrospray ionization mass spectrometry (LC-MS) along with tandem MS is described for the quantification and confirmation of N-methylpyrrolidinone (NMP) in methanolic extracts of riverine biofilm. The LC-MS method provided a 100-fold improvement in detection limits (2 ng g(-1) with a repeatability of 80-95% based on triplicate analyses) compared to a conventional LC-UV detection procedure and was applicable to quantitative analysis of biofilm samples with little or no clean up. Under low-energy collision induced dissociation (CID) conditions (17 V, laboratory frame of reference, with argon as the collision gas), two product-ions of the [M+H]+ ion were formed at m/z 69 [MH-CH3NH2]+ and m/z 58 [MH-CH3NCH]+ with relative abundances of 30% and 5%, respectively. These CID transitions were used to demonstrate that biofilm uptake of a photocatalytically-generated mixture of NMP was rapid once acclimation was achieved. PMID- 11403468 TI - Development and application of a high resolution liquid chromatographic method for the analysis of complex pigment distributions. AB - Ternary and binary gradient systems have been developed for the high-performance liquid chromatographic analysis of complex pigment distributions typical of natural samples. Improved chromatographic resolution reveals significantly more pigment components in extracts from a sediment (Priest Pot, Cumbria, UK), a microbial mat (les Salines de la Trinital, South Catalonia, Spain) and a culture (C. phaeobacteroides) including novel bacteriochlorophyll derivatives. The methods developed are directly suited to LC-MS analysis and the automated acquisition of MS/MS data for pigments. PMID- 11403469 TI - Characterization of starch polysaccharides by flow field-flow fractionation-multi angle laser light scattering-differential refractometer index. AB - The coupling between flow field-flow fractionation (FFF), multi-angle laser light scattering and differential refractometer index provides a promising technique for fractionation of starch polysaccharides in aqueous conditions. Native starches with different amylose/amylopectin levels (0-70%) as well as a pure amylose sample were characterized. By applying a sudden drop in the cross-flow rate, clear separation was achieved between amylose (which elutes first) and amylopectin. Flow FFF produced correct relationships between the molecular mass or the gyration radius versus elution volume for the fractionated amylopectin population. The results are also considered in terms of the macromolecular composition of starches. PMID- 11403470 TI - Determination of percent composition of a mixture analyzed by gas chromatography. Comparison of a helium pulsed-discharge photoionization detector with a flame ionization detector. AB - We present the results of a study of percent composition for a mixture which has been separated by gas chromatography and analyzed using helium pulsed-discharge photoionization detection (He-PDPID) and flame ionization detection (FID). FID has long been the means by which the percent composition of a hydrocarbon mixture has been determined since it has been previously established as a "carbon counting device". However, in this study we present results which show that He PDPID is more accurate in determining the percent composition of a hydrocarbon mixture and, because it is a universal detection method and can detect compounds that FID cannot, it is also more effective for determining the percent composition of mixtures containing organic compounds with a variety of other functional groups. PMID- 11403471 TI - Miniature radio-frequency mobility analyzer as a gas chromatographic detector for oxygen-containing volatile organic compounds, pheromones and other insect attractants. AB - A high electric field, radio-frequency ion mobility spectrometry (RF-IMS) analyzer was used as a small detector in gas chromatographic separations of mixtures of volatile organic compounds including alcohols, aldehydes, esters, ethers, pheromones, and other chemical attractants for insects. The detector was equipped with a 2 mCi 63Ni ion source and the drift region for ion characterization was 5 mm wide, 15 mm long and 0.5 mm high. The rate of scanning for the compensation voltages was 60 V s(-1) and permitted four to six scans to be obtained across a capillary chromatographic elution profile for each component. The RF-IMS scans were characteristic of a compound and provided a second dimension of chemical identity to chromatographic retention adding specificity in instances of co-elution. Limits of detection were 1.6-55 x 10(-11) g with an average detection limit for all chemicals of 9.4 x 10(-11) g. Response to mass was linear from 2-50 x 10(-10) g with an average sensitivity of 4 pA ng( 1). Separations of pheromones and chemical attractants for insects illustrated the distinct patterns obtained from gas chromatography with RF-IMS scans in real time and suggest an analytical utility of the RF-IMS as a small, advanced detector for on-site gas chromatographs. PMID- 11403472 TI - Characterization of stationary phases for gas chromatography by 29Si NMR spectroscopy. II. Silphenylene-siloxane copolymers. AB - A series of tetramethyl-p-silphenylene-siloxane copolymers with dimethyl, 1H,1H,2H,2H-perfluorodecylmethyl and diphenyl siloxy groups was prepared. 1H and 29Si nuclear magnetic resonance spectroscopy showed that the chosen reaction conditions provided polymers with diphenyl content up to 85%. The theoretical content of the monomer units correlated well with the measured content. Signal assignments of the copolymers and their corresponding chemical shifts are summarized. Information about alternating, randomized or block sequences was obtained by 29Si nuclear magnetic resonance spectroscopy. Limitations of the method for the determination of microstructure parameters are discussed. PMID- 11403473 TI - Suitability of several carbon sorbents for the fractionation of various sub groups of toxic polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins and polychlorinated dibenzofurans. AB - Feasibility of several sorbents, Amoco PX-21, Carbosphere, Carbopack B and C and 2-(1-pyrenyl) ethyldimethylsilylated silica gel (PYE), for the fractionation of polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins and furans was investigated. Selection was based on their suitability for quantitative isolation of the target compounds with special attention for the most toxic mono- and non ortho-CBs to prevent coelution with other congeners during the final determination by gas chromatography. Cost-effectiveness in terms of solvent and time consumption as well as feasibility for routine analysis and automation were considered additional merits of the methods compared. Final evaluation of the procedures providing the best results was done by comparison of the results obtained from the analysis of real-life samples. The results showed that, among the sorbents tested, Carbopack B and PYE were the most suitable for routine analysis. In particular, these sorbents allowed a more reliable determination of the toxic congeners and, consequently, of the toxic equivalents of 2,3,7,8 tetrachlorodibenzo-p-dioxin content in environmental samples. PMID- 11403474 TI - Setting system suitability criteria for detectability in high-performance liquid chromatography methods using signal-to-noise ratio statistical tolerance intervals. AB - For pharmaceutical products, one approach developed to assure that different chromatographic systems are capable of generating valid results is the system suitability test. Typically, a system suitability test involves numerical limits for predefined chromatographic parameters such as theoretical plates, tailing factor, injector reproducibility, etc. An estimation of the ratio of signal compared to baseline is one way to measure system performance, according to a valid method, independent of the instrument. However, since this comparison relates the height of the signal to the height of the noise, it is difficult to relate to the peak area measurements that are typically used for quantification of samples. Additionally, although peak area and peak height may be highly correlated over a wide region, peak area at very low concentrations can be more sensitive to all components of noise due to peak shape. To establish a system suitability criterion, one can use the ratio of the area signal to the baseline noise for replicate injections for samples prepared at concentration equal to the limit of quantitation during the validation studies. A lower limit for this ratio can be derived using statistical tolerance intervals. This lower limit can be applied as a system suitability criterion to measure that any system is performing adequately for measuring low level components in the sample for all future use of the method. PMID- 11403475 TI - Pressurized liquid extraction of lipids for the determination of oxysterols in egg-containing food. AB - Pressurized liquid extraction (PLE, ASE) was compared with the Folch procedure (a solid-liquid extraction with chloroform/methanol 2:1, v/v) for the lipid extraction of egg-containing food; the accuracy of PLE for the quantitative determination of oxysterols in whole egg powder was evaluated. Samples of spray dried whole egg, an Italian vanilla cake (Pandoro) and egg noodles were used. Two different extraction solvents (chloroform/methanol 2:1, v/v, and hexane/isopropanol 3:2, v/v) were tested at different extraction temperatures and pressures (60 degrees C at 15 MPa, 100 degrees C at 15 MPa, 120 degrees C at 20 MPa). No significant differences in the lipid recovery of the egg powder sample using PLE were found. However, PLE of the vanilla cake and egg noodles with the chloroform/methanol mixture was not selective enough and led to the extraction of a non-lipid fraction, including nitrogen-containing compounds. In the same samples, the pressurized hexane/isopropanol mixture gave a better recovery result, comparable to that obtained using the Folch method. Cholesterol oxidation products of the Folch extract and the pressurized liquid extract of spray dried egg powder (obtained with hexane/isopropanol 3:2, v/v, at 60 degrees C and 15 MPa) were determined by gas chromatography. PLE performed under these conditions is suitable to replace the Folch extraction, because the differences between the two methods tested were not statistically significant. Moreover, PLE shows important advantages, since the analysis time was shortened by a factor of 10, the solvent costs were reduced by 80% and the use of chlorinated solvents was avoided. PMID- 11403476 TI - On-site field sampling and analysis of fragrance from living lavender (Lavandula angustifolia L.) flowers by solid-phase microextraction coupled to gas chromatography and ion-trap mass spectrometry. AB - Solid-phase microextraction coupled to gas chromatography and mass spectrometry has been applied as a simple alternative method for the analysis of essential oil directly from lavender intact flowering spikes and genuine oils. All recognised major oil constituents were detected by this procedure, with results comparable to those given by a conventional method (organic solvent extraction). Distinctive chromatographic profiles were found for various species. PMID- 11403477 TI - Structural analysis of commercial ceramides by gas chromatography-mass spectrometry. AB - A simple method using gas chromatography-mass spectrometry was applied to analyse structures of ceramides. Identification of trimethylsilylated ceramides were obtained in short analysis times (derivatization of ceramides in 30 min at room temperature and 20 min gas chromatography mass spectrometry run) even for complex mixtures. For example in ceramide Type III, 18 peaks were observed which represent 27 various structures. The coeluted compounds were ceramides containing the same functional groups and the same carbon number but with a different distribution on the two alkyl chains of the molecule. They were accurately differentiated by mass spectrometry. Therefore, 83 structures of trimethylsilylated ceramides were identified in 11 different commercial mixtures. For 52 structures of these, mass spectral data were not described in the literature, neither full mass spectra nor characteristic fragments. PMID- 11403478 TI - Application of gas chromatography-tandem mass spectrometry to the analysis of inhibition of dimerisation of tributylphosphate under radiolysis. Identification of isomeric tributylphosphate-alkylbenzene inhibitor coupling products. AB - Tributylphosphate (TBP), solvent used as extractant for reprocessing spent nuclear fuel, can dimerise under radiolysis. This occurs by radical radical recombination, leading to 10 isomeric dimers (TBP-TBP). These species are complexation agents and are responsible of fission product retention in the organic phase that increases the solvent degradation. In order to limit their formation two free radical inhibitors (In), isopropyl and 1,4 diisopropylbenzenes, were used. These additives reduce by about 50% the concentration of TBP-TBP dimers but this reduction is not strictly followed by TBP regeneration as mixed coupling products from TBP and inhibitor are detected. By using GC-MS-MS and selectively deuterated compounds, the identification of these different isomers (TBP-In) has been realised. From these identifications and from the analysis of the proportion of the different isomers, the major primary TBP radical generated under radiolysis was determined. PMID- 11403479 TI - Miniaturized automated matrix solid-phase dispersion extraction of pesticides in fruit followed by gas chromatographic-mass spectrometric analysis. AB - In this study a simple and fast miniaturized automated matrix solid-phase dispersion method for the sample preparation and quantitative extraction of pesticides was developed and evaluated. Only 25 mg of sample and 100 microl of organic solvent were used per analysis for this new miniaturized set-up. The extracts were subsequently analysed by GC-MS without any further purification. The method was optimized for oranges and tested for the determination of a variety of organophosphorus pesticides and a pyrethroid at concentration levels below the maximum residue levels set by the European Union and authorities in The Netherlands. The limits of detection were 4-90 microg/kg. The recoveries for pesticides in orange were 83-118% and the relative standard deviations for the total procedure were 10-13% (n=4) at the limit of quantification. The feasibility of the developed method for apple, pear and grapes was also studied. Equally good results were obtained, but for apple the washing step should be omitted. PMID- 11403480 TI - Mixtures of nonionic and anionic surfactants: interactions with low-molecular mass homopeptides. AB - The interaction between low molecular-mass homopeptides and mixtures of nonionic and anionic surfactants has been assessed by using reversed-phase thin-layer chromatography. The relative strength of interaction for mixtures of sodium dodecylsulfate and tridecylalcohol diglycolate (GNX) at the molar ratios of 8:2, 6:4, 4:6 and 2:8 has been calculated and its relationship with the physicochemical parameters (number of amino acid units, hydrophobicity, side chain bulkiness, electronic characteristics) of peptides has been computed by stepwise regression analysis. Each peptide interacted with each surfactant mixture the strength of interaction markedly depending on both the character of the peptide and the composition of the surfactant mixture. The hydrophobicity and electronic properties of the amino acid units exerted the highest influence on the strength of interaction at the highest concentration of the nonionic surfactant (GNX) whereas the number of amino acid units in the peptide molecule and the bulkiness of the amino acid side chain governed the strength of interaction at the lowest concentration of GNX. PMID- 11403481 TI - New family of glutathionyl-biomimetic ligands for affinity chromatography of glutathione-recognising enzymes. AB - Three anthraquinone glutathionyl-biomimetic dye ligands, comprising as terminal biomimetic moiety glutathione analogues (glutathionesulfonic acid, S-methyl glutathione and glutathione) were synthesised and characterised. The biomimetic ligands were immobilised on agarose gel and the affinity adsorbents, together with a nonbiomimetic adsorbent bearing Cibacron Blue 3GA, were studied for their purifying ability for the glutathione-recognising enzymes, NAD+-dependent formaldehyde dehydrogenase (FaDH) from Candida boidinii, NAD(P)+-dependent glutathione reductase from S. cerevisiae (GSHR) and recombinant maize glutathione S-transferase I (GSTI). All biomimetic adsorbents showed higher purifying ability for the target enzymes compared to the nonbiomimetic adsorbent, thus demonstrating their superior effectiveness as affinity chromatography materials. In particular, the affinity adsorbent comprising as terminal biomimetic moiety glutathionesulfonic acid (BM1), exhibited the highest purifying ability for FaDH and GSTI, whereas, the affinity adsorbent comprising as terminal biomimetic moiety methyl-glutathione (BM2) exhibited the highest purifying ability for GSHR. The BM1 adsorbent was integrated in a facile two-step purification procedure for FaDH. The purified enzyme showed a specific activity equal to 79 U/mg and a single band after sodium dodecylsulfate-polyacrylamide gel electrophoresis analysis. Molecular modelling was employed to visualise the binding of BM1 with FaDH, indicating favourable positioning of the key structural features of the biomimetic dye. The anthraquinone moiety provides the driving force for the correct positioning of the glutathionyl-biomimetic moiety in the binding site. It is located deep in the active site cleft forming many favourable hydrophobic contacts with hydrophobic residues of the enzyme. The positioning of the glutathione-like biomimetic moiety is primarily achieved by the strong ionic interactions with the Zn2+ ion of FaDH and Arg 114, and by the hydrophobic contacts made with Tyr 92 and Met 140. Molecular models were also produced for the binding of BM1 and BM3 (glutathione-substituted) to GSTI. In both cases the biomimetic dye forms multiple hydrophobic interactions with the enzyme through binding to a surface pocket. While the glutathioine moiety of BM3 is predicted to bind in the crystallographically observed way, an alternative, more favourable mode seems to be responsible for the better purification results achieved with BM1. PMID- 11403482 TI - A capillary electrophoresis study on the influence of beta-cyclodextrin on the critical micelle concentration of sodium dodecyl sulfate. AB - The influence of beta-cyclodextrin (beta-CD) on the critical micelle concentration (CMC) of sodium dodecyl sulfate (SDS) was investigated by capillary electrophoresis using anionic chlorophenols as probe molecules at pH 7.0. The variations of the electrophoretic mobility of probe molecules as a function of surfactant concentration in both premicellar and micellar regions in the absence and presence of beta-CD was analyzed. The results indicate that, as a consequence of a strong inclusion complexation between beta-CD and SDS, the encapsulation of beta-CD with probe molecules is greatly diminished, or even vanished, in the presence of SDS. The complexes formed between beta-CD and SDS monomers exist predominantly in the form of a 1:1 stoichiometry, while the complexes with a 2:1 stoichiometry reported previously in the literature as a minor component may exist by less than 10%. The elevation of the CMC value of SDS depends not only on the concentration of beta-CD in the buffer electrolyte but also on methanol content in the sample solution. The binding constants of probe molecules to beta CD, to surfactant molecules, and to the complexes formed between beta-CD and SDS are reported. PMID- 11403483 TI - Determination of tobacco alkaloids in single plant cells by capillary electrophoresis. AB - A new capillary electrophoresis system with direct UV detection for the analysis of the tobacco alkaloids nicotine, nornicotine and anabasine in plant microsamples was developed. An electrolyte containing a high concentration of citric acid to provide good buffer capacity at pH 3.6 was found to be most suitable in terms of sensitivity and separation efficiency. At this low pH the tobacco alkaloids are present in cationic form, showing high mobility and increased UV absorption. This system was used for the analysis of nicotine in single epidermal leaf cells of tobacco plants. Only vacuolar concentrations of nicotine were determined, as the vacuole occupies >95% of the entire volume in epidermal cells. The procedure of sample acquisition and preparation for nicotine analysis of vacuolar samples in the pl range is shown. The results indicate a gradient of nicotine from the leaf base to the tip with higher concentrations present in the cells at the tip. Compared to simultaneously measured bulk leaf samples containing all types of cells, tissues and compartments, the concentrations in epidermal cells are much higher. As nicotine is the major defence substance against insects in tobacco and the epidermis is the most exposed leaf tissue this result is physiologically plausible. PMID- 11403484 TI - Capillary electrophoresis of methylderivatives of quinolines. I. AB - Migration behavior of quinoline, isoquinoline and related methylderivatives has been investigated with respect to the influence of running buffer acidity and to the presence of polyethylene glycol (PEG) 2000 as additive. Dissociation constants and ionic mobilities were determined by capillary electrophoresis (CE). Mobility and viscosity measurements in PEG containing buffers show that analyte transport is not in accordance with Walden's rule and microviscosity plays the role in analyte retardation. Variation of pH and PEG concentration provides the optimal conditions for the CE separation of methylquinolines (0.0176 M acetate Tris buffer, pH 5.5, 10% PEG 2000). Analysis of industrial mixture (isoquinoline fraction from distillation of coal tar) was performed and good agreement with gas chromatographic results was found. PMID- 11403485 TI - Determination of the dissociation constants of sulfonated azo dyes by capillary zone electrophoresis and spectrophotometry methods. AB - The dissociation constants of 10 sulfonated azo dyes, six of the most common food colours used as additives (Food Yellow 4, Food Yellow 3, Food Red 9, Food Red 7, Food Red 17 and Food Blue 5), and four commonly used as textile dyes (Acid Orange 7, Acid Orange 12, Acid Red 26 and Acid Red 88), have been determined by two different systems, one by using capillary electrophoresis (CE) with diode array detection and the other by using UV-visible absorption spectrophotometry, which has been used as reference method to obtain the pKa values. The pKa values obtained by CE were determined in two ways, first on the basis of the electrophoretic mobilities (calculated from the migration times), and after we propose a new methodology, in which the dissociation constants are determined from the spectra corresponding to the maxima of electrophoretic peaks. The pKa values obtained by using these CE methods have been compared with those obtained by using the spectrophotometric method. The results show that the pKa values obtained by the CE proposed method are in general closer to the reference values than those obtained from the electrophoretic mobilities. Moreover, the proposed method retains the advantages of CE, as the possibility of working with small amounts of sample, despite its purity. PMID- 11403486 TI - Optimization of the separation of a group of antifungals by capillary zone electrophoresis. AB - Two simple, rapid, and efficient methods for the analysis of seven antifungal compounds have been developed by capillary zone electrophoresis. Resolutions higher than 1.5 were obtained using 0.025 M phosphate buffer (pH 2.30) (analysis time close to 9 min) or 0.2 M formic acid (pH 2.15) (analysis time close to 6 min), with an applied voltage of 20 kV and a temperature of 30 degrees C. The highest sensitivity and selectivity can be obtained using phosphate buffer but the shortest analysis times are achieved in the formic system. The analytical characteristics of the optimized methods were investigated. The reproducibility obtained for migration times (RSD(n = 10) < or = 1.0%) and peak areas (RSD(n = 10) < or = 4.3%) was acceptable, but better reproducibilities were obtained when verapamil was used as internal standard (RSD(n = 10) < 0.4% for relative migration times and RSD(n = 10) < or = 2.2% for peak area ratios). The lowest limit of detection was obtained for clotrimazole (0.12 microg/ml) and the highest for fluconazole and voriconazole (0.90 microg/ml). The lowest and the highest limits of quantitation were, respectively, 0.40 microg/ml for clotrimazole and 3.00 microg/ml for fluconazole and voriconazole. PMID- 11403487 TI - Counter-current chromatographic separation of glycoprotein components from Morchella esculenta (L.) with a polymer phase system by a cross-axis coil planet centrifuge. AB - Using a cross-axis coil planet centrifuge, glycoproteins were separated from fermentation media of Morchella esculenta (L.) by high-speed counter-current chromatography. The performance of the apparatus was optimized with four standard proteins including pepsin, lysozyme, ovalbumin and hemoglobin and a polymer phase system composed of 12.5% (w/w) polyethylene glycol 8000 and 25% (w/w) potassium phosphate in distilled water at various pH values. Separations were performed by eluting the lower phosphate-rich phase at a flow-rate of 1.0 ml/min. Under the optimized conditions three glycoprotein components in Morchella esculenta (L.) were resolved within 6 h. PMID- 11403488 TI - Fast separation of UV absorbing anions using ion-interaction chromatography. AB - Ion-interaction chromatography on a short (30 x 4.6 mm) 3 microm ODS column has been investigated with the aim of developing fast chromatographic separations of selected inorganic anions. Tetrabutylammonium chloride (TBA-Cl) was used as the ion-interaction reagent in mobile phases that also contained up to 20% methanol. Separations of simple test mixtures of up to eight UV absorbing anions illustrated how excellent efficiencies (>50,000 plates/m) could be obtained under optimized conditions. The use of an optimised mobile phase containing 20 mM TBA Cl and 20% methanol resulted in the baseline separation of five important anions (iodate, bromate, nitrite, bromide and nitrate) in a separation window of just 28 s, with a shortest total analysis time of 50 s. The method was briefly applied to the rapid analysis of nitrite and nitrate in both a drinking water and a river water sample with a view to future on-line monitoring. PMID- 11403489 TI - Evaluation of the lipophilicity of bile acids and their derivatives by thin-layer chromatography and principal component analysis. AB - The lipophilic character of bile acids and their glyco- and tauro-conjugates was studied. The classical R(Mo) values were measured by means of reversed thin-layer chromatography using a mixture of methanol-water as the solvent system and compared with the factors scores obtained by principal component analysis based also onto the TLC-retention data. The reliability of the factor scores values as lipophilic indices are shown by their high correlation with the classical R(Mo) values. In addition, a better correlation was observed between scores corresponding to the first principal components and the partition coefficients (log P) of bile acids. Finally, the "lipophilicity chart" described by the first two components has the effect of separating compounds from each other most effectively from the congeneric aspect point of view. PMID- 11403490 TI - Investigation of the storage stability of selected volatile sulfur compounds in different sampling containers. AB - The suitability of various sample containers (i.e. standard Tedlar sample bags, black/clear layered Tedlar sample bags and Silcosteel sample cylinders) was examined for a gaseous multicomponent standard containing methylmercaptan, ethylmercaptan, dimethyl sulfide, ethylmethyl sulfide, 2-propylmercaptan, 1 propylmercaptan, 2-butylmercaptan, diethyl sulfide and 1-butylmercaptan (1 mg/m3 each in nitrogen). In the black/clear layered Tedlar sample bags, significant losses (up to 10% for methylmercaptan as the most critical component) were observed after 2 days, whereas in the standard Tedlar sample bags the recovery of methylmercaptan was approximately 90% even after 1 week. The Silcosteel sample cylinders were suitable for sampling of volatile sulfur compounds with respect to the stability of the analytes, but the recoveries exceeded 100% especially for the higher boiling compounds, which was attributed to enrichment effects on parts of the sampling system. PMID- 11403491 TI - Comparative studies on discrete and concatemeric DNA-sepharose columns for purification of transcription factors. AB - Concatemers, tandem copies of DNA elements ligated together, are widely used for the DNA affinity chromatography of transcription factors. Purification of different transcription factors using discrete, concatemeric and T18:A18 tailed DNA affinity columns was studied. Columns having a discrete DNA sequence bound by cytidylic-adenylic-adenylic-thymidylic oligonucleotide (CAAT) enhancer binding protein (C/EBP) yields significantly more green fluorescent protein-C/EBP (GFP C/EBP) fusion protein than a concatemeric DNA column made from five tandem repeats of the same DNA sequence. For lac repressor protein, the concatemeric and T18:A18 tailed columns show greater retention times than a discrete, untailed DNA affinity column. It was observed that the T18:A18 tailed column gives better resolution than either the discrete or concatemeric columns, of mixtures containing both lac repressor and GFP-C/EBP. Discrete concatemeric and T18:A18 tail columns all bound the Sp1 transcription factor and showed similar retention. The T18:A18 tailed column gives higher yield for Sp1 than the other columns. Our study shows concatemeric columns do not have any distinct advantage for the three different transcription factors we studied including Sp1, the original justification for the concatemeric approach. PMID- 11403492 TI - Bioskin as an affinity matrix for the separation of glycoproteins. AB - Bioskin is a natural product produced by a mixed culture of Acetobacter xylinum, Saccharomyces cerevisiae and S. pombe cultured on media containing sucrose. It is of fibrillar nature able to retain some proteins, such as cytochrome c, by adsorption, and mainly composed of glucosamine and N-acetyl-D-glucosamine. This makes it possible that, at an adequate pH value, proteins charged as polyanionic molecules, such as catalase, can be retained by ionic adsorption using the positively charged amino groups of the matrix. In addition, bioskin can also be used as an affinity matrix to retain glycoproteins able to perform specific affinity reactions with the amino sugars of the matrix, such as invertase, fetuin or ovalbumin. Its possible use as a chromatographic support is discussed. PMID- 11403493 TI - Investigation of pharmaceutical high-performance liquid chromatography assay bias using experimental design. AB - This article presents a systematic approach to investigate, document, and eliminate pharmaceutical HPLC assay bias using experimental design. This is the first article to describe the application of experimental design in the area of assay bias. It is found that both formulation and analytical variables can contribute to pharmaceutical HPLC assay bias using model compounds and formulations. PMID- 11403494 TI - Combination of matrix solid-phase dispersion extraction and direct on-line liquid chromatography-nuclear magnetic resonance spectroscopy-tandem mass spectrometry as a new efficient approach for the rapid screening of natural products: application to the total asterosaponin fraction of the starfish Asterias rubens. AB - A combination of matrix solid-phase dispersion extraction (MSPD) and LC-NMR-MS hyphenation is proposed as a rapid screening method of natural products for unknown compounds. In this report, this new analytical approach is applied for the first time. MSPD represents a significant simplification compared to classical extraction procedures and is thus an excellent complement to the fast and powerful LC-NMR-MS: MSPD yields extracts suitable for LC-NMR-MS in one simple preparation step, while LC-NMR-MS yields a wealth of information in one single chromatographic run. The suitability of this technique to characterise glycosidic compounds in the molecular mass range of 1200 to 1400 a.m.u. is demonstrated. The information on the number of exchangeable protons provided by an additional back exchange experiment proved to be particularly valuable for structural elucidation. The possibility of semi-quantitative LC-NMR measurements through methyl signals H(3)-18 and 19 of the steroidal skeleton is demonstrated and is ensuingly used to provide relative quantitative data of the steroid oligosaccharide fraction. PMID- 11403495 TI - Evaluation on the adsorption capabilities of new chemically modified polymeric adsorbents with protoporphyrin IX. AB - A chemically modified polymeric adsorbent was synthesized to evaluate the availability as an adsorbent for solid-phase extraction (SPE) of phenol and chlorophenols. Commercially available Amberlite XAD-2 and XAD-4 resins were modified with macrocyclic protoporphyrin IX (PPIX) through the ketone linkage. Adsorption isotherms were obtained by batch experiments and the data were fitted to the Freundlich equation to calculate the adsorption parameters. Breakthrough volumes were measured by column experiments. Physical properties such as surface area, average pore diameter and micropore volume of resins were measured to correlate with the adsorption characteristics. As a result, adsorption capacity was increased for the chemically modified resins and it can be concluded that the increase of pi-pi interaction due to the introduction of the porphyrin molecule is the major factor for the increase of the adsorption capacity. PMID- 11403496 TI - High-performance liquid chromatographic determination of methyl anthranilate, hydroxymethylfurfural and related compounds in honey. AB - A high-performance liquid chromatographic method for determining 5-hydroxymethyl 2-furaldehyde (hydroxymethylfurfural), 2-furaldehyde (furfural), furan-2 carboxylic acid (2-furoic acid), furan-3-carboxylic acid (3-furoic acid), furan-3 carboxaldehyde (3-furaldehyde) and 2-aminobenzoic acid methyl ester (methyl anthranilate) in honey and honeydew samples is described. To prevent matrix interference and to isolate the compounds, a clean-up step which implies a solid phase extraction on polymeric cartridges and an elution with 0.5 ml methanol is recommended. The compounds are separated on a reversed-phase column with a gradient of (A) 1% aqueous acetic acid-acetonitrile (97:3, v/v) and (B) acetonitrile-water (50:50, v/v), with UV detection at 250 nm. The method is applied to the analysis of samples from different botanical origin. PMID- 11403497 TI - Differential regulation of prostaglandin endoperoxide synthase-2 transcription in ovine granulosa and large luteal cells. AB - The objective of the present study was to evaluate second messenger regulation of prostaglandin synthase-2 (PGS-2) mRNA and PGS-2 promoter in ovine granulosa cells and large luteal cells. In granulosa cells, PGS-2 mRNA was induced by forskolin (PKA activator) but not by phorbol didecanoate (PDD; PKC activator) with maximal stimulation at 24 h. In contrast, PDD was the more potent inducer in large luteal cells with the most dramatic effect by 4 h. Similarly, forskolin but not PDD increased media PGF2 alpha in granulosa cells at 24 h; whereas, PDD but not forskolin increased PGF2 alpha at 4 h and 24 h in large luteal cells. To evaluate if these effects were due to activation of transcription, a plasmid was constructed containing 1500 bp of PGS-2 promoter linked to a luciferase gene. Forskolin stimulated transcription from this construct in granulosa cells (5 fold); whereas, PDD but not forskolin stimulated transcription in large luteal cells (40-fold). Taking these findings together, we propose that transcriptional regulation of the PGS-2 gene changes from PKA-dependence in granulosa cells to PKC-dependence in large luteal cells after luteinization. PMID- 11403498 TI - Plasma concentrations of estradiol 17beta and PGF2alpha metabolite and placental fatty acid composition and antioxidant enzyme activity in cows with and without retained fetal membranes. AB - Fetal membrane retention is one of the most common problems in Holstein cattle after parturition. To investigate mechanisms involved, the following parameters were studied in the peri-parturition period: plasmatic concentrations of estradiol-17beta (E2) and PGFM (PGF2alpha metabolite), activity of antioxidant enzymes (superoxide dismutase-SOD, catalase-CAT and glutathione peroxidase-GSH Px), thiobarbituric acid reagent substances (TBAR) concentrations and fatty acid composition of the placentae. E2 at parturition in the NPR group (control cows, n = 10) was higher than in PR cows (placental retention, n = 10) (P < 0.05). Activity of SOD in fetal tissue of NPR animals was higher than that of the PR group. In contrast, there was no difference between the two groups in activity of GSH-Px and CAT and the TBAR content of placental tissues. PR maternal tissues had proportionally more arachidonic and linoleic acid than tissues from NPR cows. Therefore, a complex of sequential events may cause placenta retention, starting with an unbalance of antioxidant capacity of the placenta, followed by a decrease in production of estrogen, which leads to the accumulation of arachidonic and linoleic acid in placental tissues. PMID- 11403499 TI - Platelet-activating factor (PAF)-like activity, localization of PAF receptor (PAF R) and PAF-acetylhydrolase (PAF-AH) activity in bovine endometrium at different stages of the estrous cycle and early pregnancy. AB - PAF-like activity in the endometrium increased from days 2-4 to day 12 and day 20 in both cyclic and pregnant cows. There was an increase in platelet aggregation induced by PAF-like activity in the endometrium of pregnant animals on day 20 as compared to cyclic animals at the same point in time. Two major bands of PAF-R protein at 67 kDa and 97 kDa were detected by Western blot analysis. PAF-R was localized mainly in luminal and glandular epithelium of the endometrium, but the staining was markedly increased in the endometrium of pregnant cows on day 20 compared to cyclic animals on the same day. The purified PAF-AH from the endometrium is similar to in plasma. In cyclic cattle, no changes in PAF-AH activity of endometrium were observed, whereas a decrease in enzyme activity occurred in pregnant cows on day 20 as compared to cyclic animals on the same day. We suggest that the bovine endometrium produces PAF-like activity, expresses the PAF-R and possesses a PAF-AH activity which varies during pregnancy. PMID- 11403500 TI - Novel mode of action of iloprost: in vitro down-regulation of endothelial cell adhesion molecules. AB - Iloprost is a stable prostacyclin analog commonly employed in the treatment of peripheral vascular disease and also indicated in the treatment of patients affected by systemic sclerosis (SSc) in the presence of severe Raynaud's phenomenon (RP). Several mechanisms of action of the drug other than vasodilation and antiplatelet effect have been demonstrated that may be involved in the exertion of its clinical efficacy. Aim of the present study was to investigate whether iloprost down-regulated lymphocyte adhesion to endothelium through a modulation of adhesion molecule expression on the surface of endothelial cells. In the presence of iloprost, both lymphocyte adhesion and IL-1 stimulated expression of ICAM-1 and ELAM-1 exhibited a significant reduction, while unstimulated adhesion molecule expression was not significantly affected. Our results confirm that iloprost is able to down-regulate lymphocyte adhesion to endothelial cells and indicate that endothelium itself could be target of iloprost administration. Attenuation of the inflammatory response through modulation of cellular interactions could be suggested as a potential mechanism of action of iloprost, when used in the treatment of pathological conditions characterized by endothelial activation. PMID- 11403501 TI - Oxytocin stimulates prostaglandin F2alpha secretion from porcine endometrial cells through activation of calcium-dependent protein kinase C. AB - The mechanism for oxytocin's (OT) stimulation of PGF2alpha secretion from porcine endometrium is not clear, but is thought to involve mobilization of intracellular Ca2+ and subsequent activation of protein kinase C (PKC). This study determined: (1) if mobilization of inositol trisphosphate-sensitive Ca2+ by thapsigargin or activation of PKC by phorbol 12-myristate 13-acetate (PMA) could stimulate PGF2alpha release from luminal epithelial, glandular epithelial and stromal cells of porcine endometrium and (2) if inhibitors of various PKC isotypes could attenuate the ability of OT, thapsigargin and PMA to stimulate PGF2alpha secretion from these cells. Thapsigargin and PMA each stimulated (P < 0.01) PGF2alpha secretion from all three endometrial cell types examined. However, the effects of thapsigargin and PMA were synergistic (P < 0.05) only in stromal cells. Three protein kinase C inhibitors (i.e. Go6976, Go6983 and Ro-31-8220) differentially attenuated (P < 0.05) the ability of OT, thapsigargin and PMA to stimulate PGF2alpha release. These results are consistent with the hypothesis that OT mobilizes Ca2+ to activate a Ca2+-dependent PKC pathway to promote PGF2alpha secretion from porcine endometrial cells. The differing pattern of response to isotype-specific inhibitors of PKC among cell types suggests that distinct PKC isoforms are differentially expressed in luminal epithelial, glandular epithelial and stromal cells. PMID- 11403502 TI - Secondary prevention antibiotic treatment trials. PMID- 11403503 TI - Clinical benefit of noninvasive viability studies of patients with severe ischemic left ventricular dysfunction. AB - The population of patients who have congestive heart failure of ischemic origin is large and growing. It imposes a heavy burden on human suffering and economic costs such as the chronic use of costly medications, recurrent hospital admissions, and, eventually, death or the necessity of heart transplantation. Therefore, the development of methods for detecting viable myocardium may allow the accurate selection of those patients with coronary artery disease with severe left ventricular dysfunction who are most likely to benefit from revascularization, but also excludes patients who are unlikely to obtain any improvement with revascularization techniques. The presence of reversible dysfunctional myocardium that may improve after revascularization implies the concepts of stunned and hibernating myocardium. Recent evidence suggests that hibernation may not be a stable condition since it might evolve toward an irreversible dysfunction if it is not revascularized at the right moment. The techniques available for viability studies are single-photon emission computed tomography using thallium-201 or compounds labeled with technetium-99m, positron emission tomography, and dobutamine stress echocardiography. Newer and promising techniques are magnetic resonance imaging and contrast echocardiography, whose definitive roles are not clear yet. There is abundant evidence from several important studies showing that patients with a significant amount of viable myocardium have a poor outcome if they are treated medically. Conversely, if these patients are revascularized, their outcomes improve and their symptoms significantly decrease, with less necessity of medication, fewer admissions to the hospital, and even in some cases avoiding heart transplantation. On the other hand, patients with poor or no viability who are revascularized do not obtain significant benefit. PMID- 11403504 TI - The International EECP Patient Registry (IEPR): design, methods, baseline characteristics, and acute results. AB - BACKGROUND: In 1998, the International EECP Patient Registry (IEPR) was organized to document patient characteristics, safety, and efficacy during the treatment period, and long-term outcomes. All centers with EECP facilities were invited to join the voluntary Registry. The Registry population comprises all patients starting EECP therapy for treatment of angina pectoris in participating centers. HYPOTHESIS: The study was undertaken to determine whether EECP is a safe and effective treatment for patients with angina pectoris regardless of their suitability for revascularization by more conventional techniques. METHODS: After 18 months of operation, 43 clinical centers representing over half of clinical sites using the EECP system contributed cases. The data reported here were collected before the first EECP treatment and upon completion of final treatment. EECP can be used for patients ineligible for either coronary artery bypass graft (CABG) or percutaneous coronary intervention (PCI), as well as for those who prefer noninvasive treatment to avoid or delay revascularization. In this report, patients considered to be candidates for revascularization are compared with those not considered suitable. RESULTS: Of the 978 patients analyzed, 70% had Canadian Cardiovascular Society Classification class III or IV angina before starting treatment, and 62% used nitroglycerin. Most (81%) had been previously revascularized, and 69% were considered unsuited for either PCI or CABG at the time of starting EECP. A full treatment course (usually 35 h) was completed in 86%, of whom 81% reported improvement of at least one angina class immediately after the last treatment. CONCLUSION: In a broad patient population, EECP has been shown to be a safe and effective treatment. PMID- 11403505 TI - Effect of coronary risk factors on arterial compensatory enlargement in japanese middle-aged patients with de novo single-vessel disease--an intravascular ultrasound study. AB - BACKGROUND: Compensatory enlargement (CE) of atherosclerotic human arteries has been reported; however, the pattern of arterial remodeling in response to plaque formation is not unique. HYPOTHESIS: The study was undertaken to determine the extent of coronary artery compensatory enlargement at stenotic lesions and to correlate the arterial compensatory enlargement with risk factors. METHODS: We studied 62 patients with stable angina and de novo single-vessel disease using intravascular ultrasound and obtained good images in 42 patients (68%). The vessel cross-sectional area (VA), lumen cross-sectional area (LA), and plaque cross-sectional area (PA) were measured at the lesion site and at proximal and distal reference sites. Positive CE was defined as increase in VA of lesion site > 10% compared with that of proximal reference site (CE group, n = 15); shrinkage was defined as reduction in VA of lesion site > 10% compared with that of proximal reference site (S group, n = 14); inadequate CE was defined as intermediate between CE and S (IE group, n = 13). All subjects had coronary risk factors measured before this study. RESULTS: There was no difference in VA, LA, or PA among the three groups at the proximal and distal reference sites, nor in LA at the lesion site; however, VA and PA were significantly smaller in the S group than in the other groups (p < 0.01). Of coronary risk factors, increased systolic blood pressure (SBP), increased diastolic blood pressure (DBP), and decreased high-density lipoprotein cholesterol (HDL-c) levels had the strongest association with shrinkage (p < 0.05). CONCLUSION: Hypertension and decreased HDL level may contribute to the shrinkage response in middle-aged patients with stable angina. PMID- 11403506 TI - Does higher diastolic augmentation predict clinical benefit from enhanced external counterpulsation?: Data from the International EECP Patient Registry (IEPR). AB - BACKGROUND: Enhanced external counterpulsation (EECP) has been demonstrated to be an effective treatment for stable angina in patients with coronary disease. The hemodynamic effects of EECP are maximized when the ratio of diastolic to systolic pressure area is in the range of 1.5 to 2.0. HYPOTHESIS: It is hypothesized that patients undergoing EECP who are able to achieve higher diastolic augmentation (DA) ratios may derive greater clinical benefit. This study examines the relationship between the DA ratio and clinical outcomes in patients undergoing EECP. METHODS: We analyzed demographic, noninvasive hemodynamic, and clinical outcome data on 1,004 patients enrolled in the International EECP Patient Registry (IEPR) for treatment of chronic angina between January 1998 and August 1999. Blood pressure waveforms were recorded from finger plethysmography. Six month clinical outcomes were obtained by telephone interview. RESULTS: At the end of EECP treatment, 370 (37%) patients had a higher DA ratio (defined as > or = 1.5) and 634 (63%) had a lower DA ratio (defined as < 1.5). Factors associated with a lower DA ratio included age > or =65 years (p <0.001), female gender (p < 0.001), left ventricular ejection fraction < 35% (p < 0.05), hypertension (p < 0.01), prior coronary bypass surgery (p < 0.01), noncardiac vascular disease (p < 0.001), multivessel disease (p < 0.01), congestive heart failure (p < 0.01), current smoking (p < 0.01), unsuitability for further revascularization (p < 0.001), and higher baseline angina class (p < 0.001). There were no significant differences regarding diabetes mellitus, prior coronary angioplasty, prior myocardial infarction, or antianginal medication use between patients with higher or lower DA ratios. Based on a multiple logistic regression model, independent predictors of a DA ratio < 1.5 at the end of EECP included current smoking (odds ratio 3.3; 95% confidence intervals 2.0-5.4); multivessel disease (1.7; 1.3-2.3); female gender (2.2; 1.7-3.0); no prior EECP (1.9; 1.1-3.3); noncardiac vascular disease (2.3; 1.7-2.9); age > or = 65 years (1.7; 1.4-2.2), and patients not suitable for revascularization (1.6; 1.2-2.0). By the end of therapy, there were no significant differences in myocardial infarction, revascularization rates, or nitroglycerin use with respect to higher DA ratios. At 6-month follow-up, patients with higher DA had a trend toward a greater reduction in angina class compared with those with lower DA (p = 0.069). There was a significantly higher rate of unstable angina and congestive heart failure in the group not achieving higher augmentation (p < 0.05). CONCLUSIONS: Patients who are younger, male, nonsmoking, and without multivessel coronary or noncardiac vascular disease are most likely to have higher DA with EECP. Patients with higher DA tended to have a greater reduction in angina class at 6-month follow-up compared with those with lower DA ratios. There is evidence that higher DA ratios are associated with improved short- or long-term clinical outcomes, suggesting that clinical benefit from EECP is associated with the magnitude of DA. PMID- 11403507 TI - Low clinical utility of routine angiographic surveillance in the detection and management of cardiac allograft vasculopathy in transplant recipients. AB - BACKGROUND: Cardiac allograft vasculopathy (CAV), a form of accelerated atherosclerosis, is the major cause of late death in heart transplant recipients. Routine annual coronary angiography has been used as the standard surveillance technique for CAV in most transplant centers. HYPOTHESIS: The aim of this study was to investigate the clinical utility of routine angiographic surveillance in the detection and management of CAV in transplant recipients. METHODS: We reviewed the case notes and angiograms of 230 patients who underwent cardiac transplantation in our unit between January 1986 and January 1996 and survived beyond the first year post transplantation. RESULTS: Significant complications secondary to angiography arose in 19 patients (8.2%). Cardiac allograft vasculopathy was present on none of angiograms performed 3 weeks post transplantation, but was identified in 9 patients (4%) at the first annual angiogram and an additional 25 patients by the fifth annual angiogram. A target lesion suitable for angioplasty was only identified in two patients, and only limited procedural success was achieved in both cases. Twenty-five patients (11%) died during the study period, and the most common cause of late death was graft failure which occurred in 10 patients. All patients who died from graft failure had significant CAV at autopsy, but the most recent coronary angiogram had been normal in eight of these patients. CONCLUSIONS: These data clearly illustrate the limited clinical utility of routine angiographic surveillance for CAV in heart transplant recipients and prompted us to abandon this method of surveillance in our unit. PMID- 11403508 TI - Homocysteine levels in patients with risk factors for atherosclerosis. AB - BACKGROUND: Abundant epidemiological evidence has demonstrated that the presence of mild to moderate hyperhomocysteinemia is an independent risk factor for atherosclerosis in the coronary, cerebral, and peripheral vasculature, and for vascular disease, including coronary disease. It has been demonstrated that plasma total homocysteine level is a strong predictor of mortality in patients with angiographically confirmed coronary artery disease. HYPOTHESIS: The study was undertaken to determine the extent of homocysteine levels in patients without documented coronary artery disease, but with at least one risk factor for atherosclerosis. METHODS: Fasting blood samples were collected prospectively from 160 consecutive patients (50 women and 110 men, mean age 65+/-7 years) who had at least one risk factor for atherosclerosis, but had no documented coronary artery disease. Homocysteine levels were measured by an immunoassay method. RESULTS: Of the patients studied, 78 (48.75%) with at least one risk factor for atherosclerosis had high homocysteine levels; 62 patients had mild hyperhomocysteinemia (15-30 micromol/l); and 16 patients had moderate hyperhomocysteinemia (30-100 micromol/l). CONCLUSIONS: Our data suggest that hyperhomocysteinemia is highly prevalent in patients with risk factors for atherosclerosis. Homocysteine level (an independent convertible risk factor to atherosclerosis) should be measured routinely in patients with risk factors for atherosclerosis and treated appropriately. PMID- 11403509 TI - Low-dose combination therapy with colesevelam hydrochloride and lovastatin effectively decreases low-density lipoprotein cholesterol in patients with primary hypercholesterolemia. AB - BACKGROUND: Colesevelam hydrochloride is a novel, lipid-lowering agent that binds bile acids with high affinity. A multicenter, randomized, double-blind, placebo controlled, parallel-design study was conducted to assess the efficacy and tolerability of combination low-dose colesevelam and lovastatin treatment in patients with primary hypercholesterolemia. HYPOTHESIS: Combination therapy with low doses of colesevelam and lovastatin decreases low density (LDL) cholesterol with minimal adverse events. METHODS: Following a 4- to 6-week dietary lead in, 135 patients were randomized into five groups for a 4-week treatment period: placebo, colesevelam 2.3 g at dinner, lovastatin 10 mg at dinner, the combination of colesevelam and lovastatin given at dinner (dosed together), and combination treatment with colesevelam given at dinner and lovastatin administered at bedtime (dosed apart). RESULTS: Combination colesevelam and lovastatin treatment decreased LDL cholesterol by 34% (60 mg/dl, p < 0.0001) and 32% (53 mg/dl, p < 0.0001) when colesevelam and lovastatin were dosed together or dosed apart, respectively. Both combination therapies were superior to either agent alone (p < 0.05). Decreases in LDL cholesterol exceeded the combined decreases observed for colesevelam alone (13 mg/dl, 7%) and lovastatin alone (39 mg/dl, 22%). Both combination treatments reduced total cholesterol by 21% (p < 0.0001) and apolipoprotein B by 24% (p < 0.0001). Neither combination treatment significantly altered high-density lipoprotein cholesterol or triglycerides. Adverse side effects were not significantly different among randomized groups. CONCLUSIONS: Combination colesevelam and lovastatin was efficacious and well tolerated, resulting in additive decreases in LDL cholesterol levels whether or not both agents were administered simultaneously. PMID- 11403510 TI - Women with high exercise tolerance and the role of myocardial perfusion imaging. AB - BACKGROUND: Myocardial perfusion imaging (MPI) provides incremental diagnostic and prognostic information, even in patients with high exercise tolerance. HYPOTHESIS: Myocardial perfusion imaging provides significant diagnostic value, specifically in women with high exercise tolerance. METHODS: Our study population consisted of all women who underwent exercise MPI in our Department from January 1992 to June 1996 and reached at least Stage IV in the Bruce protocol. Patients were divided into those with known and those with possible coronary artery disease (CAD). All patients were followed for 3 years from the performance of MPI. RESULTS: Of 4,803 women who underwent myocardial perfusion imaging, 3,183 had exercise stressing, and of those, 311 reached at least Stage IV in the Bruce protocol. Of these 311 MPI scans, only 23 (7.4%) were abnormal (reversible, fixed, or mixed) and the remaining 288 (92.6%) were normal. Of the 82 patients with known CAD, 13 (15.8%) had an abnormal MPI, while only 10 (4.4%) of the 229 patients with possible CAD. No myocardial infarction or cardiac death occurred within 3 years; one patient with normal MPI needed revascularization. CONCLUSION: In women with high exercise tolerance, especially in those without already known CAD, the yield of MPI is very low. Women with high exercise tolerance have an excellent prognosis. PMID- 11403511 TI - Porcelain atrium. PMID- 11403512 TI - Cardiac tamponade masking pulmonary embolism. AB - This report describes a patient admitted with shortness of breath due to cardiac tamponade, which masked concomitant pulmonary embolism that was diagnosed only after right heart pressures failed to decrease after successful pericardiocentesis. The patient was found to have widely metastatic adenocarcinoma of colon (with metastases to pericardium) and a paraneoplastic syndrome of deep vein thrombosis. PMID- 11403513 TI - William Evans. PMID- 11403514 TI - Oxidative stress in human aging and mitochondrial disease-consequences of defective mitochondrial respiration and impaired antioxidant enzyme system. AB - Respiratory function of mitochondria is compromised in aging human tissues and severely impaired in the patients with mitochondrial disease. A wide spectrum of mitochondrial DNA (mtDNA) mutations has been established to associate with mitochondrial diseases. Some of these mtDNA mutations also occur in various human tissues in an age-dependent manner. These mtDNA mutations cause defects in the respiratory chain due to impairment of the gene expression and structure of respiratory chain polypeptides that are encoded by the mitochondrial genome. Since defective mitochondria generate more reactive oxygen species (ROS) such as O2- and H2O2 via electron leak, we hypothesized that oxidative stress is a contributory factor for aging and mitochondrial disease. This hypothesis has been supported by the findings that oxidative stress and oxidative damage in tissues and culture cells are increased in elderly subjects and patients with mitochondrial diseases. Another line of supporting evidence is our recent finding that the enzyme activities of Cu,Zn-SOD, catalase and glutathione peroxidase (GPx) decrease with age in skin fibroblasts. By contrast, Mn-SOD activity increases up to 65 years of age and then slightly declines thereafter. On the other hand, we observed that the RNA, protein and activity levels of Mn-SOD are increased two- to three-fold in skin fibroblasts of the patients with CPEO syndrome but are dramatically decreased in patients with MELAS or MERRF syndrome. However, the other antioxidant enzymes did not change in the same manner. The imbalance in the expression of these antioxidant enzymes indicates that the production of ROS is in excess of their removal, which in turn may elicit an elevation of oxidative stress in the fibroblasts. Indeed, it was found that intracellular levels of H2O2 and oxidative damage to DNA and lipids in skin fibroblasts from elderly subjects or patients with mitochondrial diseases are significantly increased as compared to those of age-matched controls. Furthermore, Mn-SOD or GPx-1 gene knockout mice were found to display neurological disorders and enhanced oxidative damage similar to those observed in the patients with mitochondrial disease. These observations are reviewed in this article to support that oxidative stress elicited by defective respiratory function and impaired antioxidant enzyme system plays a key role in the pathophysiology of mitochondrial disease and human aging. PMID- 11403515 TI - The simulation effects of mountain climbing training on selected endocrine responses. AB - The simulation effects of mountain climbing exercise training on plasma testosterone, cortisol and luteinizing hormone (LH) levels were examined in ten recreational mountain male climbers. Subjects underwent a simulating mountain climbing exercise training 3 times a week for a total of eight weeks before an expedition to Mount Muztag Ata (7546 m, Xingian, China). During training, each subject carried a 40 kg back pack while walking on a treadmill at a speed of 1.9 mph for 60 min at sea level. Subjects completed an incremental treadmill test to exhaustion prior to training, after training, and one week after returning from Mount Muztag Ata. Blood samples were collected from antecubital vein at rest and at 5, 60, and 120 min post testing to determine the plasma testosterone, cortisol and LH levels. The basal plasma testosterone and cortisol concentrations were lower in both post-training and after-climbing conditions compared with that in the pre-training condition (p<0.01). The basal plasma LH concentration was remained unchanged after training and after the mountain climbing compared with levels measured in the pre-training phase. No correlation could be established between plasma LH and testosterone level. These results suggest that an eight week period of mountain climbing training protocol may be beneficial in maintaining normal endocrine function during and after high altitude mountain expedition. Our results also indicate the decrease of plasma testosterone was LH independent. PMID- 11403516 TI - Fendiline-Induced Ca2+ movement in A10 smooth muscle cells. AB - The effect of fendiline, an anti-anginal drug, on cytosolic free Ca2+ levels ([Ca2+]i) in A10 smooth muscle cells was explored by using fura-2 as a Ca2+ indicator. Fendiline at concentrations between 10-50 microM increased [Ca2+]i in a concentration-dependent manner with an EC50 of 20 microM. External Ca2+ removal reduced the Ca2+ signal by 75%. Addition of 3 mM Ca2+ increased [Ca2+]i in cells pretreated with fendiline in Ca2+-free medium. The 50 microM fendiline-induced [Ca2+]i increase in Ca2+-containing medium was inhibited by 10 microM of La3+, nifedipine, or verapamil. In Ca2+-free medium, pretreatment with 1 microM thapsigargin (an endoplasmic reticulum Ca2+ pump inhibitor) to deplete the endoplasmic reticulum Ca2+ store partly inhibited 50 microM fendiline-induced Ca2+ release; whereas pretreatment with 50 microM fendiline abolished 1 microM thapsigargin-induced Ca2+ release. Inhibition of phospholipase C activity with 2 microM U73122 did not alter 50 microM fendiline-induced Ca2+ release. Incubation with 50 microM fendiline for 10-30 min decreased cell viability by 10-20%. Together, the findings indicate that in smooth muscle cells fendiline induced [Ca2+]i increases. Fendiline acted by activating Ca2+ influx via L-type Ca2+ channels, and by releasing internal Ca2+ in a phospholipase C-independent manner. Prolonged exposure of cells to fendiline induced cell death. PMID- 11403517 TI - Neonatal chemical sympathectomy attenuates fructose-induced hypertriglyceridemia and hypertension in rats. AB - Experiments were performed to determine the pathogenic contribution of the peripheral sympathetic nervous system to fructose-induced hypertriglyceridemia, hyperinsulinemia and hypertension in rats. Neonatal chemical sympathectomy was performed in neonatal Sprague-Dawley rats (1-week old) by administration of guanethidine (50 microg/g, i.p.) 5 times per week for consecutive 3 weeks and nerve-intact rats were served as controls. Both groups of rats were fed a fructose-enriched diet for 9 weeks. The systolic blood pressure (SBP) and body weight were measured weekly and arterial blood samples were taken weekly for determinations of plasma insulin, glucose and triglyceride levels. The results showed that fructose feeding for one week significantly increased SBP in intact rats and sympathectomized rats (116+/-1 to 119+/-1 mmHg and 116+/-1 to 120+/-1 mmHg, respectively). SBP further increased thereafter in both groups. However, the increased SBP levels were significantly higher in intact group than in sympathectomized group after 5 weeks of fructose feeding. Fructose feeding for one week concurrently produced hypertriglyceridemia that preceded the appearance of hyperinsulinemia in both groups. The elevated plasma triglyceride levels were significantly lower in sympathectomized rats than in intact rats after 3 weeks of fructose feeding, whereas the elevated plasma insulin concentrations were not different between groups throughout fructose feeding period. Plasma glucose concentrations of both groups were comparable and remained unchanged throughout the study. These data indicate that neonatal chemical sympathectomy attenuated, but did not prevent, fructose-induced elevations in blood pressure and plasma triglyceride levels, suggesting a partial dependency of fructose-induced hypertriglyceridemia and hypertension on the integrity of the peripheral sympathetic nervous system (SNS) in rats. PMID- 11403518 TI - Posttraining infusion of norepinephrine and corticotropin releasing factor into the bed nucleus of the stria terminalis enhanced retention in an inhibitory avoidance task. AB - The present study investigated whether the bed nucleus of the stria terminalis (BNST) is involved in formation and retrieval of affective memory. Male Wistar rats with cannulae bilaterally implanted into the BNST were trained on a one trial step-through inhibitory avoidance task. Shortly after training they received bilateral intra-BNST infusion of lidocaine, various noradrenergic drugs, or corticotropin releasing factor (CRF). Results showed that posttraining intra BNST infusion of lidocaine impaired retention. Posttraining intra-BNST infusion of norepinephrine or the alpha1 antagonist prazosin induced a dose- and time dependent retention enhancement or deficit, respectively. The enhancing effect of norepinephrine was mimicked by the alpha1 agonist phenylephrine, and antagonized by prazosin at a non-impairing dose. Posttraining intra-BNST infusion of the alpha2 antagonist idazoxan or the beta antagonist propranolol failed to affect retention. Posttraining intra-BNST infusion of CRF also enhanced retention in a dose-dependent manner. Various drugs infused shortly before testing did not significantly influence locomotor activity and retention. These findings, taken together, suggest that the BNST is involved in memory formation processes for affective experience and norepinephrine released in the BNST acting via alpha1 receptors plays a critical role in this function. PMID- 11403519 TI - Undiagnosed diabetes mellitus in Taiwanese subjects with impaired fasting glycemia: impact of female sex, central obesity, and short stature. AB - AIMS: To study the extent and determinants of undiagnosed diabetes mellitus (DM) in Taiwanese subjects with impaired fasting glycemia (IFG) defined by the newly proposed WHO criteria. METHODS: Oral glucose tolerance tests were carried out for 306 IFG subjects identified from 6632 adult participants of two large scale community-based studies: Nutrition and Health Survey in Taiwan (1993-1996) and the Cardiovascular Disease Two-township Study (1994-1997). Similar protocols were used in these two studies to collect data on glycemic status, anthropometric measurements, and other data used in the present study. RESULTS: Subjects with IFG had a non-trivial undiagnosed diabetes mellitus rate (30% in men and 42% in women) and a high rate of glucose intolerance and undiagnosed DM combined (75% in men and 86% in women). Women with IFG had a 1.6 fold higher risk (p = 0.04) for undiagnosed DM and a 2.1 fold higher risk (p = 0.01) for glucose intolerance and DM when compared to men. There were more women than men with an elevated body mass index in undiagnosed DM patients. Among IFG subjects, undiagnosed DM patients were significantly (p < 0.05) older, more centrally obese and shorter than their normal IFG counterparts, irrespective of gender. In men, height was independent of age and waist circumference in predicting undiagnosed DM (p = 0.037). CONCLUSIONS: A high proportion of impaired glucose tolerance and undiagnosed DM was found in subjects with IFG. Its public health impact should not be overlooked. Central obesity, female sex, and short stature were associated with undiagnosed DM status in IFG subjects. PMID- 11403520 TI - Effects of pinealectomy on baseline sleep and response to sleep deprivation. AB - STUDY OBJECTIVES: We have previously reported that older (24 mo.) Fischer rats manifest a diminished post-sleep deprivation increase in NREM and REM sleep. In order to examine whether this decline reflects an age-related change in pineal function, we are now reporting on baseline and recovery sleep parameters in pinealectomized 3-, 12-, and 24-month old rats following 24 hours of sleep deprivation using the disk-over-water method. DESIGN: Three independent age groups; within each group there were sequential measures of sleep under baseline conditions and during recovery from sleep deprivation. SETTING: The Sleep Research Laboratory at the University of Chicago PARTICIPANTS: 56 male Fisher (F344) rats INTERVENTIONS: 24 hours of total sleep deprivation using the disk over-water method MEASUREMENTS: Sleep staging of EEG and EMG, and power spectral analysis of the EEG RESULTS: Pinealectomized (pinex) rats did not differ from sham-operated (sham) rats in total sleep, REM sleep, super-modal high-amplitude NREM sleep (HS2), a measure of NREM EEG delta power, or circadian rhythm amplitude. In the pinex rats, there was a modest (2.5%) age-independent increase in NREM sleep (p<0.02). The pinex rats of all ages failed to manifest the increase in NREM sleep during recovery seen in the sham-operated animals (p<0.04). CONCLUSIONS: We found no evidence that altered pineal function is responsible for age-related changes in baseline sleep in the rat. These data also suggest that, independent of age, normal pineal function may be relevant to the ability to generate increased NREM sleep in response to prior sleep deprivation. PMID- 11403521 TI - EEG bands during wakefulness, slow-wave, and paradoxical sleep as a result of principal component analysis in the rat. AB - Rat EEG has been empirically divided in bands that frequently do not correspond with EEG generators nor with the functional meaning of EEG rhythms. Power spectra from wakefulness (W), slow-wave sleep (SWS), and paradoxical sleep (PS) of Wistar rats were submitted to Principal Component Analyses (PCA) to investigate which frequencies are covariant. Three independent eigenvectors were identified for SWS: a band between 1-6, an intermediate band between 7-15, and a fast band between 16-32 Hz (90.74% of the variance); two independent eigenvectors were extracted for PS: slow frequencies between 1-6 covarying together with frequencies between 11-16 Hz, and activity between 6-10 covarying together with fast frequencies between 17-32 Hz (80.38% of the variance); four eigen-vectors were obtained for W: 3-7, 8-9, 10-21 and 21-32 Hz (81.47% of the variance). Vigilance states showed significant differences in AP from 1 to 22 Hz. PCA extracted broad bands different for each vigilance state, which included the most representative EEG activities characteristic of them. These results indicate that during SWS, slow oscillations include frequencies up to 6 Hz, and spindle oscillations frequencies down to 7 Hz. No alpha frequencies were identified as an independent band. Frequencies within theta and beta were gathered in the same eigenvector during PS and in different eigenvectors during W suggesting coordinated activation of hippocampal and cortical systems during PS. These bands are consistent with the underlying neurophysiological mechanisms of sleep and wakefulness and with firing frequencies of generators of rhythmic activity obtained in cellular studies in animals. PMID- 11403522 TI - A comparison of three different sleep schedules for reducing daytime sleepiness in narcolepsy. AB - STUDY OBJECTIVE: To determine if the combination of scheduled sleep periods and stimulant medications were more effective than stimulant medications alone in controlling the excessive daytime sleepiness experienced by narcoleptic patients. DESIGN: Twenty-nine treated narcoleptic subjects were randomly assigned to one of three treatment groups: 1) two 15-minute naps per day; 2) a regular schedule for nocturnal sleep; or 3) a combination of scheduled naps and regular bedtimes. Measures of symptom severity and unscheduled daytime were obtained at baseline and at the end of the two-week treatment period, using the Narcolepsy Symptom Status Questionnaire (NSSQ) and 24-hour ambulatory polysomnographic monitoring. No alterations were made in stimulant medications during the study period. SETTING: N/A. PATIENTS OR PARTICIPANTS: N/A. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: The addition of two-15 minute naps did not alter either symptom severity or the duration of unscheduled daytime sleep. Regular times for nocturnal sleep reduced perceived symptom severity, but did not reduce the amount of unscheduled daytime sleep. Only the combination of scheduled naps and regular nocturnal sleep times, significantly reduced both symptom severity and the amount of unscheduled daytime sleep in treated narcoleptic subjects. The type of sleep schedule prescribed, however, was less important than the severity of the patients' pre treatment daytime sleepiness. Subjects with severe daytime sleepiness benefited from the addition of scheduled sleep periods, while those who were only moderately sleepy or able to maintain alertness did not benefit from scheduled sleep periods. CONCLUSIONS: Scheduled sleep periods are helpful for only those patients who remain profoundly sleepy despite stimulant medications and should not be prescribed for all patients with narcolepsy. PMID- 11403523 TI - Insufficient sleep--a population-based study in adults. AB - STUDY OBJECTIVES: Insufficient sleep (sleep deprivation) is a common problem of considerable health, social, and economical impact. We assessed its prevalence and associations, and the role of genetic influences. DESIGN: Panel study based on questionnaires administered in 1981 and 1990. SETTING/PATIENTS: 12.423 subjects aged 33-60 years included in the Finnish Twin Cohort, representative of the Finnish population. INTERVENTIONS: N/A. MEASUREMENTS: A difference of 1 hour between the self-reports of the sleep need and the sleep length was considered insufficient sleep. Associations with education, life style, work, psychological characteristics and sleep-wake variables were assessed. Structural equation modelling techniques were used to compare genetic models among monozygotic and dizygotic twin pairs. RESULTS: In 1990, the prevalence of insufficient sleep was 20.4% (16.2% in men and 23.9% in women). 44% of those with insufficient sleep in 1981 also had it 9 years later (Spearman correlation for persistence 0.334). In multivariate analyses, the strongest positively associated factors were daytime sleepiness (women: odds ratio 3.87, 95% confidence limits 3.24-4.63/men: 3.77, 2.98-4.75), insomnia (2.48, 1.92-3.19/2.91, 2.17-3.90), not able to sleep without disturbance (1.95, 1.47-2.60/2.54, 1.66-3.89), and evening type (2.10, 1.65 2.69/1.73, 1.25-2.41). Among men, also weekly working hours > or =75 was strongly associated (3.23, 1.54-6.78). "Not working" was negatively associated in both genders (0.68, 0.51-0.89/0.59, 0.42-0.83). Two thirds of the interindividual variability in the liability to insufficient sleep was attributed to non-genetic factors. CONCLUSIONS: Insufficient sleep is a common and long-standing condition, most strongly associated with sleep/wake variables. One third of the liability to it is attributed to genetic influences. Sleep sufficiency should be assesssed in health examinations of working adults. PMID- 11403524 TI - Night driving, season, and the risk of highway accidents. AB - Official accident and traffic density statistics on Swedish highways were used to compute the relative risk (Odds Ratio - OR) of being injured or killed in a traffic accident at different times of day. After removing accidents due to alcohol 10344 accidents remained for computations, and the period 10:00h-11:00h was used as the reference point. The highest total risk was seen at 0400h (OR=5.7, Confidence interval = 5.6-5.8), with an OR of 11.4 (Ci=10.3-12.5) for fatal accidents at the same point. The same pattern was exhibited by single vehicle, head-on, and "other" (e.g., turning off the road) accidents, whereas overtaking and rear-end accidents did not show clear 24 hour patterns. Retaining alcohol-related accidents approximately doubled the nighttime peak for total accidents. During the winter, the peak of total accidents occurred at 03:00h (OR=3.8, Ci=3.5-4.0), five hours before sunrise, whereas the summer peak occurred at 04:00h (OR=11.6, Ci=11.3-11.9), shortly after the early summer sunrise and with consistently higher nighttime risk than for winter driving. It was concluded that early morning driving is several times more dangerous than driving during the forenoon. Apart from alcohol the effect seems related to sleepiness, but not to darkness. PMID- 11403525 TI - Treatment preference and patient satisfaction in chronic insomnia. AB - STUDY OBJECTIVES: The purposes of this study were to examine treatment preference and satisfaction with group treatment in individual with chronic insomnia. DESIGN: Correlational. SETTING: The study was conducted in an outpatient hospital setting. PATIENTS OR PARTICIPANTS: Participants were 43 adult volunteers from the community. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Prior to treatment, participants were presented with descriptions of behavioral and pharmacological treatment for the problem of insomnia and asked to rate the acceptability, presumed effectiveness, and presumed side-effects of treatment. A sub-sample of these individuals (n = 37) participated in a 6-week cognitive behavioral treatment group for insomnia. Sleep diary and questionnaire data were collected prior to and following treatment. Results showed that cognitive-behavioral therapy was significantly preferred over pharmacological therapy at pre-treatment and that more favorable assessments of cognitive-behavioral therapy at pre treatment were associated with better adherence but not improved outcome. Of treatment techniques, participants least liked sleep restriction and most liked sleep hygiene. Results indicated that more favorable ratings of the usefulness of sleep restriction were associated with improvements in sleep efficiency, sleep related impairment, and quality of life. CONCLUSIONS: Implications of these findings are that patient preference is important to assess prior to treating insomnia and that more work may be needed to increase patients' awareness of the benefits of sleep restriction. PMID- 11403526 TI - Melatonin in older people with age-related sleep maintenance problems: a comparison with age matched normal sleepers. AB - STUDY OBJECTIVES: To determine whether older people with age-related sleep maintenance problems have significantly lower melatonin levels than comparable normal sleepers. DESIGN: Case-control study. SETTING: A largely urban population, Auckland, New Zealand. PARTICIPANTS: People over the age of 65 years, who either slept normally, or had age-related sleep maintenance problems. Participants were recruited through media advertising, and local interest groups. Initial screening was by mail (Pittsburgh Sleep Quality Index), followed by interviews at a hospital day clinic. Exclusions included those with depression, cognitive impairment, medical and/or environmental problems which might impair sleep. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: A metabolite of plasma melatonin, 6 sulphatoxymelatonin (aMT6s) was measured in the urine of 57 normal sleepers, and 53 people with age-related problems over 24 hours in three aliquots: 12:00 19:00h, 19:00-07:00h, 07:00-12:00h. There were clear differences in self reported quality of sleep but no difference in mean aMT6s 24 hour or total night excretory levels, or night/day ratios. CONCLUSIONS: Older people with age-related sleep maintenance problems do not have lower melatonin levels than older people reporting normal sleep. PMID- 11403527 TI - Insomnia in men-a 10-year prospective population based study. AB - STUDY OBJECTIVES: to prospectively analyze changes in the prevalence of insomnia and the relationship between insomnia, aging, lifestyle, and medical disorders DESIGN: a longitudinal population survey. PARTICIPANTS: a randomly selected population sample of 2,602 men (age 30-69 years) from Uppsala in Sweden. INTERVENTION: all participants answered a questionnaire on sleep disturbances, lifestyle factors, and medical disorders in 1984 and again in 1994. MEASUREMENTS AND RESULTS: The prevalence of INSOMNIA was 10.3% in 1984 and 12.8% in 1994. No significant correlation was found between age and insomnia in any of the two time periods. Insomnia in 1994 was independently related to having insomnia in 1984 (OR=6.45), being over-weight (BMI> 27 kg/m2) (OR=1.35), physical inactivity (OR=1.42), alcohol dependence (OR=1.75), psychiatric disorders (OR=8.27) and joint/low back disorders (OR=2.95). The number of subject with reported insomnia in 1984 but not 1994 was 149. Subjects that quit smoking during the time period had an increased likeliness of remission (OR=2.70) while men who were overweight were less likely to remit (OR=0.43). CONCLUSIONS: We conclude that in men insomnia is related to lifestyle factors such as obesity, physical inactivity and alcohol dependency but not to aging. Medical disorders such as joint and low back disorders and psychiatric illnesses also increase the risk of reporting insomnia. This study demonstrates the close relationship between quality of sleep and overall health status. PMID- 11403528 TI - In vivo efficacy of two heated humidifiers used during CPAP-therapy for obstructive sleep apnea under various environmental conditions. AB - STUDY OBJECTIVES: To investigate the humidification performance-defined as the maximum achievable absolute humidity in the CPAP tube-of two heated humidifier systems (HH) offered as CPAP accessories, as a function of ambient air conditions. DESIGN: In 48 patients undergoing CPAP treatment, temperature (T) and relative humidity (RH) in the distal CPAP tube system were measured, with and without either of the two heated humidifiers A (HH-A, n=23), or B (HH-B, n=25), until a steady state was achieved. At the same time, ambient T and RH in the examination room were recorded. T and RH were used to calculate the absolute humidity (AH). SETTING: University Hospital, Erlangen, Germany. PATICIPANTS: 48 patients with obstructive sleep apnea undergoing CPAP therapy. INTERVENTIONS: N/A. MEASUREMENTS AND RESULTS: Conditions in the examination room during measurement with the HH-A, T = 22.5+2.1 (16.4-26.0) degrees C and AH = 9.3+2.4 (5.3-13.9) g/m3 did not differ significantly from those prevailing during measurements with the HH-B, T = 22.9+1.9 (18.9-26.3) degrees C and AH = 9.9+2.8 (6.2-16.4) g/m3. The mean humidification performance (steady state AH with HH within the CPAP tube) of the HH-A was 23.5+2.9 (19.1-29.9) g/m3, that of the HH-B 26.8+3.9 (21.0-34.4) g/m3. CONCLUSIONS: Under the ambient conditions of humidity and temperature, commonly found in European and North American bedrooms, both HH demonstrate a high humidification performance that even falls within the range recommended for intubated patients. The difference between the two HH is small, and probably not clinical relevant. Thus, it would appear that both HH are suitable for the treatment of dry upper airways under CPAP therapy. PMID- 11403529 TI - Arousal components which differentiate the MWT from the MSLT. AB - STUDY OBJECTIVES: The purpose of this study was to determine the relative contribution of the instruction to maintain wakefulness versus posture change as major components determining sleep latency in the MWT as compared to the MSLT. DESIGN AND SETTING: After adaptation, subjects spent 3 nights and the following days in the laboratory. On each day, Ss had eight sleep latency measurements including four sleep latency tests from two of the following conditions: Lay down and Sleep (MSLT); Lay down and stay Awake; Sit up and Sleep; Sit up and stay Awake (MWT); and sit in a chair in front of a Computer and stay awake. PARTICIPANTS: Participants were 14 young adult normal sleepers. INTERVENTIONS: NA. MEASUREMENT AND RESULTS: Significant differences in sleep latency were found for each condition with respect to all of the others except that the Computer condition did not differ from the Sit-Awake condition. Means for conditions were: Lay-Sleep -11.1 minutes; Sit-Sleep -17.7 minutes; Lay-Awake - 21.7 minutes; Sit Awake - 29.0 minutes; Computer - 30.1 minutes. Correlations between conditions declined as subjects sat up. CONCLUSIONS: The MWT differs from the MSLT by taking advantage of the arousal system (motivation and posture) to maintain alertness (i.e., increase sleep latency). These arousal effects are additive. MSLT results may not always correlate well with MWT results because the MWT measures the combined effects of the sleep and arousal systems while the MSLT, in ideal situations, measures only sleepiness. PMID- 11403530 TI - Practice parameters for the treatment of narcolepsy: an update for 2000. AB - Successful treatment of narcolepsy requires an accurate diagnosis to exclude patients with other sleep disorders, which have different treatments, and to avoid unnecessary complications of drug treatment. Treatment objectives should be tailored to individual circumstances. Modafinil, amphetamine, methamphetamine, dextroamphetamine, methylphenidate, selegiline, pemoline, tricyclic antidepressants, and fluoxetine are effective treatments for narcolepsy, but the quality of published clinical evidence supporting them varies. Scheduled naps can be beneficial to combat sleepiness, but naps seldom suffice as primary therapy. Regular follow up of patients with narcolepsy is necessary to educate patients and their families, monitor for complications of therapy and emergent of other sleep disorders, and help the patient adapt to the disease. PMID- 11403531 TI - Hypopnea in sleep-disordered breathing in adults. PMID- 11403532 TI - Pharmacological and pharmacodynamic essentials of H(2)-receptor antagonists and proton pump inhibitors for the practising physician. AB - The suppression of gastric acid secretion with anti-secretory agents has been the mainstay of medical treatment for patients with acid-related disorders. Although the majority of Helicobacter pylori -related peptic ulcers can be healed with antibiotics, ulcer healing and symptom control can be significantly improved when antibiotics are given with anti-secretory agents, especially with a proton pump inhibitor. There is a dynamic relationship between the suppression of intragastric acidity and the healing of peptic ulcer and erosive oesophagitis and control of acid-related symptoms. The suppression of gastric acid secretion achieved with H(2)-receptor antagonists has, however, proved to be suboptimal for effectively controlling acid-related disorders, especially for healing erosive oesophagitis and for the relief of reflux symptoms. H(2)-receptor antagonists are also not effective in inhibiting meal-stimulated acid secretion, which is required for managing patients with erosive oesophagitis. Furthermore, the rapid development of tolerance to H(2)-receptor antagonists and the rebound acid hypersecretion after the withdrawal of an H(2)-receptor antagonist further limit their clinical use. Although low-dose H(2)-receptor antagonists are currently available as over-the-counter medications for self-controlling acid-related symptoms, their pharmacology and pharmacodynamics have not been well studied, especially in the self-medicating population. Proton pump inhibitors have been proved to be very effective for suppressing intragastric acidity to all known stimuli, although variations exist in the rapidity of onset of action and the potency of acid inhibition after oral administration at the approved therapeutic doses, which may have important clinical implications for the treatment of gastro oesophageal reflux disease and perhaps for eradicating H. pylori infection when a proton pump inhibitor is given with antibiotics. Once-daily dosing in the morning is more effective than dosing in the evening for all proton pump inhibitors with respect to the suppression of intragastric acidity and daytime gastric acid secretion in particular, which may result from a better bio-availability being achieved with the morning dose. When higher doses are needed, these drugs must be given twice daily to achieve the optimal suppression of 24 hour intragastric acidity. Preliminary results have shown that esomeprazole, the optical isomer of omeprazole, given at 40 mg, is significantly more effective than omeprazole 40 mg, lansoprazole 30 mg or pantoprazole 40 mg for suppressing gastric acid secretion. However, more studies in different patient populations are needed to compare esomeprazole with the existing proton pump inhibitors with regard to their efficacy, cost-effectiveness and long-term safety for the management of acid-related disorders. PMID- 11403533 TI - Histamine receptor antagonists, proton pump inhibitors and their combination in the treatment of gastro-oesophageal reflux disease. AB - The medical treatment of gastro-oesophageal reflux disease is accomplished with the appropriate use of anti-secretory therapy, principally H(2)-receptor antagonists and proton pump inhibitors. In fact, there is a direct correlation between the length of time, in terms of the number of hours per day that the intragastric pH is above 4, and the healing of the oesophagitis. Nowadays, H(2) receptor antagonists are of limited use as primary treatment, being inferior to proton pump inhibitors in both healing and symptom relief. Although the majority of patients can be effectively managed with carefully titrated doses of proton pump inhibitors, a small number will continue to show difficulty in the management of their disease, principally because of inadequate nocturnal acid control. These patients may benefit from a combination of proton pump inhibitors twice daily with an H(2)-receptor antagonist at bedtime. This article reviews the use of H(2)-antagonists, proton pump inhibitors and their combination in the management of the patient with gastro-oesophageal reflux disease. PMID- 11403534 TI - Current indications for acid suppressants in dyspepsia. AB - Anti-secretory drug use is common in patients with uninvestigated and functional dyspepsia, but the value of such agents has been controversial. Four large studies have evaluated the symptomatic outcome after a short course of acid inhibition in patients with uninvestigated dyspepsia presenting in primary care. All of these studies demonstrated a superior symptom response to proton pump inhibitor therapy compared with placebo and acid-alginates or H(2)-receptor antagonists. In patients with documented functional dyspepsia, 17 parallel group trials have evaluated an H(2)-receptor antagonist against placebo, with mixed results. A recent Cochrane review based on eight controlled trials concluded that there was a significant benefit of H(2)-blockers over placebo with a relative risk reduction of 30%, but as gastro-oesophageal reflux disease was not excluded, the conclusions are questionable. Six controlled studies have compared symptom relief after a short course of proton pump inhibitor therapy compared with placebo. Overall, there does appear to be a therapeutic gain with proton pump inhibitors over placebo, although how much of this is explained by undiagnosed gastro-oesophageal reflux disease remains unclear. There is conflicting evidence on the value of symptom subgrouping as a predictor of response to acid suppression. Overall, there is little convincing evidence that Helicobacter pylori infection influences the therapeutic outcome of acid-suppressant therapy. In conclusion, there appears to be a subgroup of patients with functional dyspepsia who will respond to acid suppression over and above placebo, but further work is required to characterize these responders. PMID- 11403535 TI - Current indications for acid suppressants in Helicobacter pylori -negative ulcer disease. AB - Although Helicobacter pylori infection remains the single most common cause of peptic ulcer, an increasing proportion of patients have H. pylori -negative ulcers. The proportion is higher in the USA--and possibly Australia--than elsewhere. Although the precise aetiology of these ulcers is often unknown, some are caused by the use of aspirin or non-steroidal anti-inflammatory drugs. In areas with a high prevalence of H. pylori -negative ulcers, the empirical treatment of H. pylori infection for newly diagnosed peptic ulcer disease should be discouraged. All such patients should have documentation of their H. pylori status before treatment. Patients with H. pylori -negative ulcers may have the more serious ulcer diathesis and are likely to require long-term management with acid-suppressing drugs. Proton pump inhibitors are likely to be the drugs of choice; patients may be relatively refractory to H(2)-receptor antagonists. The optimal duration of treatment is undefined but might be lifelong. There are no prospective studies of the efficacy of surgery or mucosal-protective agents in the treatment of H. pylori -negative ulcers. PMID- 11403536 TI - Current role of acid suppressants in Helicobacter pylori eradication therapy. AB - Helicobacter pylori induces chronic active gastritis that may progress to atrophy. Serious clinical consequences are peptic ulcer disease and gastric malignancies. Today, treatment of the infection is an appropriate option and is strongly recommended in various clinical situations. Although many antibiotics are effective against H. pylori in vitro, few substances are suitable for use in vivo. This is because H. pylori lives in a unique environment in which several factors may affect the pharmacokinetic and pharmacodynamic properties of the anti microbial agents. One of the most important factors is gastric acidity. This article reviews the effects of acid suppression on H. pylori and the associated gastritis, the potential mechanisms by which anti-secretory drugs such as proton pump inhibitors might enhance the activity of anti-microbials in vivo, and the results of clinical trials supporting the current view that proton pump inhibitors are a mainstay in the treatment of this infection. PMID- 11403537 TI - Role of acid suppressants in prophylaxis of NSAID damage. AB - Gastric acid contributes to the pathogenesis of non-steroidal anti-inflammatory drug (NSAID)-induced ulceration via several mechanisms, including conversion of superficial to deeper injury, impairment of haemostasis, and interference with ulcer healing. The suppression of acid secretion has been shown to reduce the severity of NSAID-induced mucosal damage in experimental models and clinical studies. Current evidence indicates that proton pump inhibitors (PPIs) are the preferred treatment for the healing of gastric ulcers when NSAIDs cannot be discontinued. PPIs are superior to standard-dose H(2)-receptor antagonists and equivalent to low-dose misoprostol in preventing NSAID-induced gastric ulcers. Whether there is any significant advantage of PPIs over higher doses of H(2) receptor antagonists or misoprostol is unknown. The efficacy of PPIs is enhanced in the presence of H. Pylori infection. Omeprazole has been shown to be effective for the secondary prevention of ulcer bleeding in H. pylori -infected NSAID users. The efficacy of PPIs for the prevention of ulcer complications in H. pylori-negative NSAID users remains uncertain. PMID- 11403538 TI - Role of acid suppressants in intensive care medicine. AB - Bleeding from stress-induced mucosal lesions continues to be a potential problem in critically ill patients, although its incidence has decreased dramatically over the past decade. Patients considered to be at risk are those with respiratory failure, coagulopathy, severe burns or tetraplegia. The most important cause of stress ulcer bleeding is tissue hypoxia. Provided that appropriate dosage regimens are administered, all agents approved for stress ulcer prophylaxis may reduce the incidence of overt as well as clinically important bleeding. However, the efficacy of stress ulcer prophylaxis does not correlate with the efficacy of gastric acid inhibition. Although numerous studies have demonstrated that an alkaline gastric juice is associated with gastric Gram negative bacterial overgrowth, controversy remains over whether the pharmacological suppression of gastric acid in critically ill patients facilitates nosocomial pneumonia. The reasons for these divergent results are discussed, as is a possible association between gastric acid suppression and other systemic infections. Finally, several cost-effectiveness analyses performed over recent years have demonstrated that, in properly selected critically ill patients, stress ulcer prophylaxis is cost-effective. PMID- 11403539 TI - The role of acid suppressants in upper gastrointestinal ulcer bleeding. AB - Re-bleeding and mortality remain significant in peptic ulcer haemorrhage despite the widespread use of endoscopic therapy. The acidic gastric environment interferes with coagulation. In vitro studies show that an intragastric pH of above 6 results in normal blood coagulation and platelet function. Because of induced tachyphylaxis, H(2)-receptor antagonists are not able to maintain a high pH. In addition, in randomized trials using H(2)-receptor antagonists, there was no reduction in re-bleeding and mortality. High-dose intravenous proton pump inhibitors are capable of maintaining a pH above 6. Four randomized trials, using high-dose intravenous proton pump inhibitors, significantly improved the outcome (in terms of a reduction in re-bleeding and surgery) in patients with peptic ulcer haemorrhage. Mortality was, however, not reduced. The additional effect of acid-suppressant agents after successful endoscopic therapy is limited to the reduction of re-bleeding and need for surgery, with no effect on mortality. PMID- 11403540 TI - Gastric acid suppression and treatment of severe exocrine pancreatic insufficiency. AB - Adding either H(2)-receptor antagonists (cimetidine or ranitidine) or proton pump inhibitors to an adequate amount of lipolytic activity improves fat malabsorption in most cases and abolishes steatorrhoea in up to 40% of children and adults with cystic fibrosis and in adults with chronic pancreatitis. Acid suppression improves fat absorption because the resultant increase in pH within the upper gastrointestinal tract improves the survival of lipolytic activity, reduces duodenal volume flow and prevents the precipitation of bile acids. These effects increase the concentration of intraduodenal lipolytic activity and promote the aggregation of bile acids and the micellar solubilization of lipid. The amount of lipase that should be recommended is controversial, but we interpret our studies as indicating that at least 90 000 United States Pharmacopeia (USP) units should be ingested with meals. This amount of lipolytic activity taken with an agent that suppresses gastric acid secretion improves fat absorption in most patients and may even abolish steatorrhoea. PMID- 11403541 TI - Problems related to acid rebound and tachyphylaxis. AB - Rebound acid hypersecretion after taking histamine H(2)-receptor antagonists is a now well-established class phenomenon. It has been demonstrated both basally and in response to meal and gastrin-releasing peptide stimulation, but not in response to peak pentagastrin stimulation. It is present by 3 days after treatment but has resolved by 10 days. A recent study in previously asymptomatic healthy volunteers has suggested that this phenomenon may be clinically relevant. Tachyphylaxis/tolerance after the use of H(2)-receptor antagonists is also now well established. It manifests as a loss of acid inhibitory efficacy and is also a class effect. It is present within a few doses but is not progressive after 29 days. Rebound acid hypersecretion after proton pump inhibitors has been shown for both basal and maximal acid output by 14 days after treatment. It is found in Helicobacter pylori -negative, but not positive, subjects, probably owing to the influence of the enhanced oxyntic gastritis that occurs during proton pump inhibitor therapy. It is a prolonged phenomenon, lasting for at least 2 months after a 2-month treatment course. This duration is likely to reflect its development as a result of trophic effects on the oxyntic mucosa. This trophism is caused by the marked hypergastrinaemia that occurs secondary to the profound acid suppression during proton pump inhibitor treatment. The clinical relevance of this phenomenon remains at present unknown. Tachyphylaxis/tolerance has not yet been shown in several short-term studies after taking proton pump inhibitors. A recent clinical study has, however, suggested that this phenomenon may merit longer-term evaluation. PMID- 11403542 TI - Gastric mucosal morphological consequences of acid suppression: a balanced view. AB - In the chapter, an analysis of the literature on the relationship between Helicobacter pylori, the use of proton pump inhibitors and the development of atrophic gastritis is presented, and the difficulties of classifying gastritis and the new possibilities of quantifying chronic inflammation by morphometric analysis are discussed. The issue surrounding the necessity of eradicating H. pylori in H. pylori-positive patients has still not been solved. Most studies have now accepted that proton pump inhibitors indeed accelerate the onset of atrophic gastritis in H. pylori-positive patients, but evidence against such an association was published in one recent (Scandinavian) study; conclusions from this study have, however, been challenged by several groups. Some data are available on the efficacy of H. pylori eradication with regard to the prevention of atrophy. The limited significance of the development of parietal cell protrusions and fundic gland cysts is better understood, but much less is known of the development and long-term consequence of H. pylori-induced autoimmune gastritis. Finally, recent studies in H. pylori-positive patients indicate that treatment with proton pump inhibitors may promote bacterial N-nitrosation formation. These data taken together suggest that the eradication of H. pylori may be based not only on morphological arguments, but also on bacterial alterations in the gastric milieu. PMID- 11403543 TI - Occurrence and significance of gastric colonization during acid-inhibitory therapy. AB - There are now a wide variety of drugs available that are able profoundly to reduce the production of gastric acid. These drugs are currently widely prescribed for the treatment of peptic ulceration and gastro-oesophageal reflux disease. One of the main functions of gastric acid is to kill ingested bacteria. Colonization of the gastric lumen occurs in patients on anti-secretory medication, the degree of bacterial overgrowth depending upon the degree of elevation of the pH. There have been concerns that these bacteria may produce carcinogenic nitrosamines and increase the risk of gastric cancer, but there is at present no definitive evidence in support of this. A profound suppression of gastric acid may also facilitate the colonization of the upper small intestine, leading to deconjugation of the bile salts and malabsorption. There is some evidence that profound gastric acid suppression may decrease the number of ingested pathogens required to produce enteric disease. This chapter discusses these potential bacterial complications of therapeutic acid suppression and the evidence for them. PMID- 11403544 TI - Alterations in intragastric nitrite and vitamin C levels during acid inhibitory therapy. AB - Most nitrite entering the healthy acid-secreting stomach is derived from dietary nitrate. The latter is absorbed from the small intestine, 25% then being secreted by the salivary glands into the mouth. Buccal organisms subsequently convert 20% of this nitrate to nitrite. When this nitrite is swallowed, the ascorbic acid in the acidic gastric juice reduces it to nitric oxide, which is absorbed by the mucosa. In the process, the ascorbic acid is oxidized to dehydroascorbic acid. When the intragastric pH is elevated by powerful anti-secretory agents, this gastric chemistry is profoundly modified. At a neutral pH, the swallowed nitrite does not react with ascorbic acid but accumulates in the stomach. The level of nitrite in the gastric juice during treatment with anti-secretory medication is particularly high after a nitrate-containing meal. Powerful anti-secretory medication also lowers the intragastric concentration of ascorbic acid and total vitamin C, probably because of the relative instability of the vitamin at a higher pH. These changes in the intragastric concentrations of nitrite and ascorbic acid are most marked in Helicobacter pylori -infected subjects on proton pump inhibitor therapy. It is recognized that an elevated nitrite-to-ascorbic acid ratio predisposes to the formation of potentially carcinogenic N -nitroso compounds. It is, however, unclear at present whether such compounds are formed within the human stomach. PMID- 11403545 TI - Alteration in digestion and absorption of nutrients during profound acid suppression. AB - Gastric acid suppression therapy has for many years been the cornerstone of the treatment of peptic disease. The availability of more potent inhibitors of gastric acid secretion and the increasing demand for maintenance therapy has renewed interest in the potential side-effects of profound and/or long-lasting therapy. This chapter focuses on the potential interference of gastric acid suppression therapy with the process of the digestion and absorption of nutrients. The theoretical mechanisms by which hypochlorhydria resulting from gastric acid suppression therapy may hamper digestion and absorption are multiple and well documented. Clinical studies evaluating the effect of gastric acid suppression therapy on the assimilation of nutrients are, on the other hand, scarce and have, moreover, yielded conflicting results. The reason for the latter may be related, at least in part, to elements of study design. Data indicating overt malabsorption or clear deficiencies in patients on long-term gastric acid suppression therapy are currently lacking. Nevertheless, it seems prudent, while awaiting the results of additional long-term studies, regularly to monitor these patients, especially those with increased nutrient demand, poor intake or suboptimal stores. PMID- 11403549 TI - Discussion: The Eurocleft Project 1996-2000. PMID- 11403550 TI - Levatorplasty, a new technique to treat hypernasality: anatomical investigations and preliminary clinical results. AB - Background: Velopharyngoplasty is the most commonly used operative technique for the treatment of velopharyngeal insufficiency. By attaching a posterior pharyngeal flap to the velum, a nonphysiological situation is created in the upper airway. Purpose: The aim of this investigation was to find a new surgical approach to physiological reconstruction of the velopharyngeal sphincter. Material: Anatomical investigations were performed in four cadavers. From this study a new surgical technique was developed and called 'levatorplasty': the musculus longus capitis was taken to create a new muscular loop leading to (a) an augmentation of the posterior wall, (b) a medial shift of the lateral pharyngeal wall; and (c) stretching of the velum posteriorly. Thus, the velopharyngeal space was reduced and a physiological closure of the nasal airway space could be obtained. Study design: The levatorplasty was employed in nine cleft palate patients with velopharyngeal insufficiency. Pre- and postoperatively the velopharyngeal closure was evaluated by phonetic and radiological examination. Results: The operation was easily performed without major complications. A concentric constriction with decrease of the velopharyngeal space was achieved and a definitive decrease of nasalance and hypernasality resulted. Conclusion: Long-term follow-ups have to verify whether these results will be stable. They also have to be compared with functional improvements following velopharyngoplasty or pharyngoplasty. Of special interest will be evaluation of the altered mobility of the lateral pharyngeal walls. Copyright 2001 European Association for Cranio-Maxillofacial Surgery. PMID- 11403551 TI - Cyclopia: a radiological and anatomical craniofacial post mortem study. AB - Cyclopia is a rare foetal malformation characterized by a single palpebral fissure and a proboscis associated with severe brain malformation. Approximately 1.05 in 100 000 births including stillbirths are identified as cyclopian. Cyclopia is not compatible with life. The authors present an anatomical and histological study of the fronto-orbito-maxillary region carried out after 3-D CT reconstruction in a 21 week-old foetus with cyclopia. Anatomical and histological observations suggest that the integrity of the trigeminal nerves is very important for the normal development of the embryological structures of the face. Fusion of the facial processes in the midline takes place even if central prosencephalic structures are absent. For this reason the face in cyclopia, in both its positive and negative aspects, constitutes a model for the study of the normal development of this region. Copyright 2001 European Association for Cranio Maxillofacial Surgery. PMID- 11403552 TI - Computer-aided 3-D simulation and prediction of craniofacial surgery: a new approach. AB - Background: In plastic and reconstructive craniofacial surgery, careful preoperative planning is essential. In complex cases of craniofacial synostosis, rapid prototyping models are used to simulate the surgery and reduce operating time. Recently, 3-D CT model surgery has been introduced for presurgical planning and prediction of the postoperative result. Objective: For simulation of craniofacial surgery a computer-based system was developed that allows visualization and manipulation of CT-data using computer graphics techniques. Surgical procedures in all areas of the bony skull can be performed interactively. Results: The case of a child with scaphocephalus is presented. Surgery is planned using the craniofacial surgery simulator described above. Conclusion: The computer-based interactive surgery simulation systems presented here allow precise visualization of craniofacial surgery. The accurate computer aided 3-D simulation of bone displacements is also the prerequisite for transfer of the simulated surgery using a navigation system for surgery. Thus the preoperatively planned procedure could be transferred directly to the operating table. Copyright 2001 European Association for Cranio-Maxillofacial Surgery. PMID- 11403553 TI - The fronto-orbital osteotomy as plastic-reconstructive approach to the anterior and middle skull base. AB - Introduction: A combined extra-intracranial access for the operative exploration of tumours of the anterior and middle skull base is indicated when the tumour extends intracranially and simultaneously into the nasal cavity, the paranasal sinuses or the orbit. Methods: Two standardized modifications of the fronto orbital osteotomy, the fronto-orbito-nasal and the fronto-orbito-zygomatic osteotomy, allow safe removal of skull base tumours in these locations. In extensive skull base tumours, a modified bilateral fronto-orbital-zygomatic osteotomy can be used. Results: Between February 1993 and July 2000 skull base tumours in 111 patients were resected using the presented methods. The most frequent tumour type was meningioma in 29 cases. Complications were encountered in 13 cases (11.7%). Conclusion: The advantages over other approaches are good extra- and intracranial overview and minimal cerebral trauma. Additional transfacial incisions are not usually necessary. Exact repositioning of the fronto-orbital segments leads to optimal aesthetic results. Copyright 2001 European Association for Cranio-Maxillofacial Surgery. PMID- 11403554 TI - Original sagittal split osteotomy revisited for mandibular distraction. AB - Introduction: A malformed mandible and an abnormally positioned mandibular foramen make it difficult to plan an ideal osteotomy line for mandibular distraction. In addition, there have been reports of such complications as nonunion, damage and stretch injury of the inferior alveolar nerve and tooth germ damage when conventional osteotomy or corticotomy are used for mandibular distraction. The authors utilized the original sagittal split ramus osteotomy for mandibular distraction. Patients and Methods: Five patients (three unilateral hemifacial microsomia, one bilateral hemifacial microsomia, and one mandibular retrusion) were included in this study of distraction osteogenesis using the sagittal split ramus osteotomy. Extraoral distraction devices were applied to the first four patients. An intraoral device with mono-cortical screw fixation was used for the fifth patient. Result: In all five cases, the results of the distraction were satisfactory. Complications (as listed) of conventional osteotomy when used for distraction were avoided. Satisfactory results were achieved and these were also well maintained postoperatively (mean follow up: 36 months). Conclusion: The authors believe that sagittal osteotomy for mandibular distraction osteogenesis makes it possible, to avoid injury to the inferior alveolar nerve during operation and stretching injury during distraction and to prevent tooth germ injury. It is also possible to diversify the osteotomy line for various force vectors to enlarge the bony contact surface area. Therefore, we suggest that sagittal split ramus osteotomy should be used as a preferred modification of osteotomy for mandibular distraction. Copyright 2001 European Association for Cranio-Maxillofacial Surgery. PMID- 11403555 TI - A complication of submandibular intubation in a panfacial fracture patient. AB - We present the complication of a mucocele in the floor of the mouth caused by a submandibular intubation in a patient with a panfacial fracture. Copyright 2001 European Association for Cranio-Maxillofacial Surgery. PMID- 11403556 TI - Depression, anxiety and quality of life among scheduled cosmetic surgery patients: multicentre prospective study. AB - Objective: To measure cosmetic surgery patients' state of psychological vulnerability. Method: A multicentre prospective study was carried out in three hospital departments. One hundred and three patients scheduled for cosmetic surgery were examined using a structured interview and using three assessment scales: the MADRS (Montgomery and Asberg Depression Rating Scale), the SISST (Social Interaction Self Statement Test) and the EQ-5D (EuroQol) which measures quality of life. Results: The MADRS index was higher than that of the control group (p<0.01) with 20% depressive patients. SISST: the social anxiety was greater than that of the control group (p<0.001). The EQ-5D visual analogue scale average was 77.39% indicating that there was no significant difference when compared with the control group, but the descriptive EQ-5D revealed an over representation of the anxiety/depression category (p<0.01); 50% had already taken psychotropic treatment of which 27% were an antidepressant. Conclusion: The cosmetic surgery population presents a significant state of psychological vulnerability. Copyright 2001 European Association for Cranio-Maxillofacial Surgery. PMID- 11403557 TI - Reasons for patients' discontent and litigation. AB - Introduction: Discontent and litigation among patients is a problem which increasingly preoccupies the medical profession. Aim: We aim to analyse the origin of discontent and litigation and to help avoiding these claims. Material: One hundred and seventy-eight medical expert opinions were evaluated, all made following examination of the complainant. Methods: Depending on the results of the clinical examination and the study of the files it was determined whether there was either a case of malpractice or insufficient informed consent, or no fault at all in a legal sense. In addition the patient and the surgeon were questioned as to their point of view regarding the procedure and their communication and relationship before and after treatment. Results: Frequent complaints were pain (either during treatment or afterwards), major swelling or bleeding, disturbances of trigeminal or facial nerve function, poor scar formation, loss of teeth or fixtures, faulty occlusion and discrepancies between the expected and the actual result of treatment. In 26 cases actual faults made during medical treatment were discovered. In 49 further cases, poor explanation of the proposed procedure was the reason for complaint. In the majority of remaining cases, neither faulty treatment nor insufficient information given to the patient lead to the complaint but the patient's expectations were unrealistically high. Conclusion: A considerable proportion of lawsuits originate from misunderstandings, and not treatment errors: The surgeons often concentrate on the legal requirements of informed consent and neglect to explain the practical consequences of the operation; the patients in turn tend not to ask about possible complications. Copyright 2001 European Association for Cranio Maxillofacial Surgery. PMID- 11403558 TI - Letter to the Editor. PMID- 11403559 TI - In reply. PMID- 11403560 TI - Robust oscillations within the interlocked feedback model of Drosophila circadian rhythm. AB - A mechanism for generating circadian rhythms has been of major interest in recent years. After the discovery of per and tim, a model with a simple feedback loop involving per and tim has been proposed. However, it is recognized that the simple feedback model cannot account for phenotypes generated by various mutants. A recent report by Glossop, Lyons & Hardin [Science286, 766 (1999)] on Drosophila suggests involvement of another feedback loop by dClk that is interlocked with per-tim feedback loop. In order to examine whether interlocked feedback loops can be a basic mechanism for circadian rhythms, a mathematical model was created and examined. Through extensive simulation and mathematical analysis, it was revealed that the interlocked feedback model accounts for the observations that are not explained by the simple feedback model. Moreover, the interlocked feedback model has robust properties in oscillations. PMID- 11403561 TI - A new method for estimating environmental variability for clonal organisms, and the use of fluctuating asymmetry as an indicator of developmental Instability. PMID- 11403562 TI - Local thermodynamic stability scores are well represented by a non-central student's t distribution. AB - Local folding in mRNAs is closely associated w ith biological functions. In this study, we reveal the whole distribution of local thermodynamic stability in the complete genome of the poliovirus P3/Leon/37 and the single-stranded RNA sequences that corresponds to the nucleotide sequence of the complete genome sequence (1 667 867 bp) of Helicobacter pylori (H. pylori) strain 26695. Local thermodynamic stability in the RNA sequences is measured by two standard z scores, significance score and stability score. To estimate the distribution of thermodynamic stability, a model based on the non-central Student's t distribution has been developed. Significant patterns of extremes that are either much more stable or unstable than expected by chance are detected. Our results indicate that the highly stable and statistically more significant folding regions are predominantly in non-coding sequences in the two genome sequences. Moreover, the highly unstable folding regions, on the contrary, are predominantly in the protein coding sequences of H. pylori. The observed differences across the complete genomic sequences are statistically very significant by a chi2-test. These extreme patterns may be useful in searching for target sequences for long chain antisense RNA and for locating potential RNA functional elements involved in the regulation of gene expression including translation, mRNA localization and metabolism. PMID- 11403563 TI - Adaptive value of polymorphism in intracellular self/not-self discrimination? AB - A microbial pathogen species can adapt to its host species to the extent that members of the host species are uniform. Loss of this uniformity would make it difficult for a pathogen species to transfer, from one member of the host species to another, what it had "learned" through selection of its members with advantageous mutations. The existence of major histocompatibility complex (MHC) polymorphism indicates that non-uniformity within a species is an effective host defence strategy. By virtue of this molecular discontinuity among its members the host species can "present a moving target" to the pathogen. Many proteins other than MHC proteins show polymorphism - a phenomenon which has suggested that mutations in regions of protein molecules which do not affect overt function are neutral. However, in the context of the author's differential aggregation theory of intracellular self/not-self discrimination as previously applied to the problem of the antigenicity of cancer cells, such polymorphism should serve for the recruitment of subsets of self-antigens into the antigenic repertoire of an infected cell. These would act as "intracellular antibodies" by virtue of their weak, but specific, aggregation with pathogen proteins. Peptides from the self antigens, as well as (or instead of) those from the antigens of the pathogen, would then serve as targets for attack by cytotoxic T cells. Thus, polymorphism of intracellular proteins should be of adaptive value, serving to amplify and individualize the immune response to intracellular pathogens. PMID- 11403564 TI - Pair approximation for lattice models with multiple interaction scales. AB - Pair approximation has frequently proved effective for deriving qualitative information about lattice-based stochastic spatial models for population, epidemic and evolutionary dynamics. Pair approximation is a moment closure method in which the mean-field description is supplemented by approximate equations for the frequencies of neighbor-site pairs of each possible type. A limitation of pair approximation relative to moment closure for continuous space models is that all modes of interaction between individuals (e.g., dispersal of offspring, competition, or disease transmission) are assumed to operate over a single spatial scale determined by the size of the interaction neighborhood. In this paper I present a multiscale pair approximation which allows different sized neighborhoods for each type of interaction. To illustrate and test the approximation I consider a spatial single-species logistic model in which offspring are dispersed across a birth neighborhood and established individuals have a death rate depending on the population density in a competition neighborhood, with one of these neighborhoods nested inside the other. Analysis of the steady-state equations yields several qualitative predictions that are confirmed by simulations of the model, and numerical solutions of the dynamic equations provide a close approximation to the transient behavior of the stochastic model on a large lattice. The multiscale pair approximation thus provides a useful intermediate between the standard pair approximation for a single interaction neighborhood, and a complete set of moment equations for more spatially detailed models. PMID- 11403565 TI - ESS analysis of mechanistic models for territoriality: the value of scent marks in spatial resource partitioning. AB - In this paper elements of game theory are used to analyse a spatially explicit home range model for interacting wolf packs. The model consists of a system of nonlinear partial differential equations whose parameters reflect the movement behavior of individuals within each pack and whose solutions describe the patterns of space-use by each pack. By modifying the behavioral parameters, packs adjust their patterns of movement so as to maximize their reproductive output. This involves a tradeoff between maximizing prey intake and minimizing conflict with neighbors. Evolutionarily stable choices of the behavioral parameters yields territories that are immune to invasion by groups with alternate behaviors. PMID- 11403566 TI - Computational fluid dynamics in the oral cavity of ram suspension-feeding fishes. AB - We have modeled steady, three-dimensional flow with a no-slip boundary condition in cylindrical and conical oral cavities possessing vertical or slanted branchial slits. These numerical simulations illustrate the transport of food particles toward the esophagus, as well as the velocity profiles of water exiting the oral cavity via the branchial slits. The maximum and average velocities are highest for flow exiting the most posterior branchial slit. The highest volume flow rates also occur in the most posterior slit for the cylindrical simulations, but occur in the most anterior slit for the conical simulations. Along the midline, there is a pronounced bilaterally symmetrical vortex in the posterodorsal region of the cylindrical and conical oral cavities and a second bilaterally symmetrical vortex in the posteroventral region of the cylinder. Particles entrained in the vortices will recirculate in the posterior oral cavity, increasing the probability of encounter with sticky, mucus-covered surfaces such as the oral roof, gill arches, or gill rakers. The posterodorsal vortex could serve to concentrate particles near the entrances of the epibranchial organs. The ventral vortex could be involved in sequestering dense inorganic particles that sink toward the floor of the oral cavity. All vortices are absent in the conical simulation with vertical branchial slits, indicating that the slanted branchial slits between the gill arches are responsible for the formation of the vortex in the conical oral cavity. Experiments using in vivo flow visualization techniques are needed to determine whether ram suspension feeders, pump suspension feeders, and non suspension-feeding fishes possess vortices in the posterior oral cavity that contribute to particle transport, food particle encounter with sticky surfaces, and inorganic particle rejection. PMID- 11403567 TI - Integrative study on chromosome evolution of mammals, ants and wasps based on the minimum interaction theory. AB - There is well-known evidence that in many eukaryotes, different species have different karyotypes (e.g. n=1-47 in ants and n=3-51 in mammals). Alternative (fusion and fission) hypotheses have been proposed to interpret this chromosomal diversity. Although the former has long been accepted, accumulating molecular genetics evidence seems to support the latter. We investigated this problem from a stochastic viewpoint using the Monte Carlo simulation method under the minimum interaction theory. We found that the results of simulations consistently interpreted the chromosomal diversity observed in mammals, ants and wasps, and concluded that chromosome evolution tends to evolve as a whole toward increasing chromosome numbers by centric fission. Accordingly, our results support the fission hypothesis. We discussed the process of chromosome evolution based on the latest theory of the molecular structure of chromosomes, and reconfirmed that the fission burst is the prime motive force in long-term chromosome evolution, and is effective in minimizing the genetic risks due to deleterious reciprocal translocations and in increasing the potential of genetic divergence. Centric fusion plays a biological role in eliminating heterochromatin (C-bands), but is only a local reverse flow in contrast to the previously held views. PMID- 11403569 TI - Blot overlays with 32P-labeled fusion proteins. AB - Proteins labeled with 32P can be used as sensitive "prime" in blot overlays to detect binding proteins or domains. Small G-protein Ras can bind GTP with extremely high affinity (Kd approximately 10(-11)-10(-12) M) in the presence of Mg2+. We have taken advantage of this property of Ras to develop a vector that expresses proteins of interest such as glutathione S-transferase (GST)/Ras fusion proteins for noncovalent labeling with [gamma-32P]GTP. The labeling efficiency of this method is >60% and involves a single short incubation step. We have previously identified several binding proteins for the second SH3 domain of the adaptor Nck using this method. Here we illustrate the overlay method using the GST/Ras system and compare results with the SH3 domain labeled by phosphorylation with [gamma-32P]ATP. Both methods are similarly specific and sensitive; however, we show that signals are dependent primarily on GST-mediated probe dimerization. These dimeric probes allow a more stable probe-target complex similar to immunoglobulin interactions, thus significantly improving the sensitivity of the technique. PMID- 11403570 TI - Using the yeast interaction trap and other two-hybrid-based approaches to study protein-protein interactions. AB - The detection of physical interaction between two or more molecules of interest can be facilitated if the act of association between the interactive partners leads to the production of a readily observed biological or physical readout. Many interacting molecule pairs (X, Y) can be made to induce such a readout if X and Y are each fused to defined protein elements with desired properties. For example, in the yeast forward two-hybrid system, X is synthesized as a translational fusion to a DNA-binding domain (DBD), Y is synthesized as a fusion to a transcriptional activation domain (AD), and coexpression of DBD-X and AD-Y induces transcription of easily scored responsive reporters. Other approaches use paradigms based on the artificial production of two, hybrid, molecules, but substitute a variety of readouts including the repression of transcription, activation of signal transduction pathways, or reconstitution of a disrupted enzymatic activity. In this article, we summarize a number of two-hybrid-based approaches, and detail the use of the forward yeast two-hybrid system in a screen to identify novel interacting partners for a protein of interest. PMID- 11403571 TI - The tandem affinity purification (TAP) method: a general procedure of protein complex purification. AB - Identification of components present in biological complexes requires their purification to near homogeneity. Methods of purification vary from protein to protein, making it impossible to design a general purification strategy valid for all cases. We have developed the tandem affinity purification (TAP) method as a tool that allows rapid purification under native conditions of complexes, even when expressed at their natural level. Prior knowledge of complex composition or function is not required. The TAP method requires fusion of the TAP tag, either N or C-terminally, to the target protein of interest. Starting from a relatively small number of cells, active macromolecular complexes can be isolated and used for multiple applications. Variations of the method to specifically purify complexes containing two given components or to subtract undesired complexes can easily be implemented. The TAP method was initially developed in yeast but can be successfully adapted to various organisms. Its simplicity, high yield, and wide applicability make the TAP method a very useful procedure for protein purification and proteome exploration. PMID- 11403572 TI - Mass spectrometry for the study of protein-protein interactions. AB - The identification of subpicomolar amounts of protein by mass spectrometry (MS) coupled with two-dimensional methods to separate complex protein mixtures is fueling the field of proteomics, and making feasible the notion of cataloging and comparing all of the expressed proteins in a biological sample. Functional proteomics is a complementary effort aimed at the characterization of functional features of proteins, such as their interactions with other proteins. Proteins comprise modular domains, many of which are noncatalytic modules that direct protein-protein interactions. Capturing proteins of interest and their interacting proteins by using high-affinity antibodies presents a simple method to prepare relatively simple protein mixtures easily resolved in one-dimensional formats. Individual or mixtures of proteins identified as stained bands in polyacrylamide gels are subjected to in situ digestion with the protease trypsin, and the extracted peptide fragments are analyzed by MS. The quality, quantity, and complexity of the tryptic digest, the species origin of the proteins, and the quality of the corresponding databases of genomic and protein information greatly influence the subsequent MS analysis in terms of degree of difficulty and methodological approach required to make an unambiguous protein identification. In this article we report the isolation of associated proteins from a complex cell-derived lysate by using an epitope-directed antibody. The protein pICLn engineered to carry an epitope tag was purified from cultured human embryonic kidney cells, and found to associate with a variety of proteins including the spliceosomal proteins smE and smG. By application of this general approach, the systematic identification of protein complexes and assignment of protein function are possible. PMID- 11403573 TI - Screening phage-displayed combinatorial peptide libraries. AB - Among the many techniques available to investigators interested in mapping protein-protein interactions is phage display. With a modest amount of effort, time, and cost, one can select peptide ligands to a wide array of targets from phage-display combinatorial peptide libraries. In this article, protocols and examples are provided to guide scientists who wish to identify peptide ligands to their favorite proteins. PMID- 11403574 TI - TAT-mediated protein transduction into mammalian cells. AB - Manipulation of mammalian cells has been achieved by the transfection of expression vectors, microinjection, or diffusion of peptidyl mimetics. While these approaches have been somewhat successful, the classic manipulation methods are not easily regulated and can be laborious. One approach to circumvent these problems is the use of HIV TAT-mediated protein transduction. Although this technology was originally described in 1988, few improvements were reported in the subsequent 10 years. In the last few years, significant steps have been taken to advance this technology into a broadly applicable method that allows for the rapid introduction of full-length proteins into primary and transformed cells. The technology requires the synthesis of a fusion protein, linking the TAT transduction domain to the molecule of interest using a bacterial expression vector, followed by the purification of this fusion protein under either soluble or denaturing conditions. The purified fusion protein can be directly added to mammalian cell culture or injected in vivo into mice. Protein transduction occurs in a concentration-dependent manner, achieving maximum intracellular concentrations in less than 5 min, with nearly equal intracellular concentrations between all cells in the transduced population. Full-length TAT fusion proteins have been used to address a number of biological questions, relating to cell cycle progression, apoptosis, and cellular architecture. Described here are the fundamental requirements for the creation, isolation, and utilization of TAT fusion proteins to affect mammalian cells. A detailed protocol for production and transduction of TAT-Cdc42 into primary cells is given to illustrate the technique. PMID- 11403575 TI - Intein-mediated ligation and cyclization of expressed proteins. AB - Protein splicing is a posttranslational processing event that releases an internal protein sequence from a protein precursor. During the splicing process the internal protein sequence, termed an intein, embedded in the protein precursor self-catalyzes its excision and the ligation of the flanking protein regions, termed exteins. The dissection of the splicing pathway, which involves the precise cleavage and formation of peptide bonds, and the identification of key catalytic residues at the splice junctions have led to the modulation of the protein splicing process as a protein engineering tool. Novel strategies have been developed to use intein-catalyzed reactions for the production and manipulation of proteins and peptides. These new approaches have broken down the size limitation barrier of chemical synthetic methods and are less technically demanding. The purpose of this article is to describe how to use self-splicing inteins in protein semisynthesis and backbone cyclization. The first two sections of the article provide a brief review of the distinct chemical steps that underlie protein splicing and intein enabled technology. PMID- 11403576 TI - Application of fluorescence resonance energy transfer to the GroEL-GroES chaperonin reaction. AB - Fluorescence resonance energy transfer (FRET) is a sensitive and flexible method for studying protein-protein interactions. Here it is applied to the GroEL-GroES chaperonin system to examine the ATP-driven dynamics that underlie protein folding by this chaperone. Relying on the known structures of GroEL and GroES, sites for attachment of fluorescent probes are designed into the sequence of both proteins. Because these sites are brought close in space when GroEL and GroES form a complex, excitation energy can pass from a donor to an acceptor chromophore by FRET. While in ideal circumstances FRET can be used to measure distances, significant population heterogeneity in the donor-to-acceptor distances in the GroEL-GroES complex makes distance determination difficult. This is due to incomplete labeling of these large, oligomeric proteins and to their rotational symmetry. It is shown, however, that FRET can still be used to follow protein-protein interaction dynamics even in a case such as this, where distance measurements are either not practical or not meaningful. In this way, the FRET signal is used as a simple proximity sensor to score the interaction between GroEL and GroES. Similarly, FRET can also be used to follow interactions between GroEL and a fluorescently labeled substrate polypeptide. Thus, while knowledge of molecular structure aids enormously in the design of FRET experiments, structural information is not necessarily required if the aim is to measure the thermodynamics or kinetics of a protein interaction event by following changes in the binding proximity of two components. PMID- 11403577 TI - Imaging protein-protein interactions using fluorescence resonance energy transfer microscopy. AB - Fluorescence resonance energy transfer (FRET) detects the proximity of fluorescently labeled molecules over distances >100 A. When performed in a fluorescence microscope, FRET can be used to map protein-protein interactions in vivo. We here describe a FRET microscopy method that can be used to determine whether proteins that are colocalized at the level of light microscopy interact with one another. This method can be implemented using digital microscopy systems such as a confocal microscope or a wide-field fluorescence microscope coupled to a charge-coupled device (CCD) camera. It is readily applied to samples prepared with standard immunofluorescence techniques using antibodies labeled with fluorescent dyes that act as a donor and acceptor pair for FRET. Energy transfer efficiencies are quantified based on the release of quenching of donor fluorescence due to FRET, measured by comparing the intensity of donor fluorescence before and after complete photobleaching of the acceptor. As described, this method uses Cy3 and Cy5 as the donor and acceptor fluorophores, but can be adapted for other FRET pairs including cyan fluorescent protein and yellow fluorescent protein. PMID- 11403578 TI - High-throughput yeast two-hybrid assays for large-scale protein interaction mapping. AB - Protein-protein interactions play fundamental roles in many biological processes. Hence, protein interaction mapping is becoming a well-established functional genomics approach to generate functional annotations for predicted proteins that so far have remained uncharacterized. The yeast two-hybrid system is currently one of the most standardized protein interaction mapping techniques. Here, we describe the protocols for a semiautomated, high-throughput, Gal4-based yeast two hybrid system. PMID- 11403579 TI - The value of scholarship. PMID- 11403580 TI - Unpacking the 'value added' impact of continuing professional education: a multi method case study approach. AB - The tangible benefits of continued professional education (CPE) continues to be widely debated within nursing. Whilst there is a plethora of literature relating to CPE, there is an absence of empirical evidence confirming the value of educational activities. Using a case study approach, this study sought to examine the relationship between undertaking a named course (Children's Neuroscience Course) and the perceived benefits to practitioners. Nursing students who had successfully undertaken the course (n = 14) were invited to participate. Self report evaluations (completed at induction, mid point and exit), academic performance, and data collected from a questionnaire were analyzed using non parametric measures. In addition, semi-structured interviews (n = 9) were undertaken and analyzed using a thematic content approach. Second level analysis involved triangulating the various data sets using coding frames and three themes were identified: improved knowledge, improved care delivery and professional development, which were judged as benefits to the practitioners who had undertaken the course. Further, there was a significant relationship between undertaking the course and participant's perception of their increased abilities in delivering care to the child with a neurological problem and their families. This paper will illustrate the benefits of a multi-method case study approach for evaluating the 'value added' impact of educational provision. PMID- 11403581 TI - Clinical staff as mentors in pre-registration undergraduate nursing education: students' perceptions of the mentors' roles and responsibilities. AB - This paper reports part of a multiple-phase action research initiated by a university nursing department in Hong Kong. Local hospitals were invited to collaborate in a joint mentoring programme which recruited clinical staff to mentor pre-registration nursing students during clinical placement. Interviews based on the five roles -assisting, befriending, guiding, advising and counselling, as outlined by the English National Board, were conducted to explore students' views of the mentor's roles. Findings showed that students in general agreed that the five roles depicted were necessary roles of the mentors. They saw the roles of assisting and guiding as most crucial, while the befriending role was useful to facilitate their settling into the ward. The advising and counselling roles were seen as less important. It was concluded that students tended to be more instrumental in viewing their mentor's behaviours; activities that have a direct effect on their learning are considered as more important, while other factors which have a less immediate or less direct effect on their learning, are seen as of lower priority. PMID- 11403582 TI - The role of the personal tutor: a literature review. AB - This paper comprises an extensive review of the literature surrounding the role of the personal tutor, which was conducted prior to an evaluation of the personal tutor system used within my own institution. There is agreement in the literature that student support is an important function of the nurse lecturer, but there is currently no consensus as to the most appropriate system of providing this support. Three distinct aspects of the personal tutor role emerged from the literature and are discussed in this paper:* Clinical role* Pastoral role* Academic role. Few research studies were identified in the literature search; this highlights the need for nurse lecturers to evaluate the effectiveness of support systems currently in place. There is also a need to undertake further research studies, in order to provide the evidence needed by nurse lecturers to determine the most appropriate model for student support. PMID- 11403583 TI - 'Real life' clinical learning on an interprofessional training ward. AB - This paper describes the multi-method evaluation of an interprofessional training ward placement for medical, nursing, occupational therapy and physiotherapy students. Unique in the UK, and an extension of pioneering work in Sweden (Wahlstrom et al. 1997, Wahlstroom & Sanden 1998), this interprofessional clinical placement allowed senior pre-qualifying students, under the supervision of practitioners, to plan and deliver interprofessional care for a group of orthopaedic and rheumatology patients. This responsibility enabled students to develop both their profession-specific skills in a real-world setting and the quality of their interprofessional teamwork. Student teams were supported by facilitators who led reflective sessions and acted as a resource for the students' problem-based learning. The training ward was evaluated by a multi method approach, incorporating interviews, observations and questionnaires with students, patients and clinical staff. The evaluation findings have been grouped into a number of themes which offer an insight into the varying perspectives of training ward students, patients and staff. This paper pays particular attention to the nursing perspective of the interprofessional training ward pilot. PMID- 11403584 TI - Teaching planning and reflection in nurse education. PMID- 11403585 TI - The value of intelligent multimedia simulation for teaching clinical decision making skills. AB - This paper examines the value of using intelligent multimedia simulation for the teaching of nursing clinical decision-making skills. The possibilities of multimedia-based educational resources are examined and the rapid growth and questionable effectiveness of current multimedia computer-based learning applications for nursing students are discussed. The advantages and disadvantages of this technology and the problems developing intelligent agent-based systems are examined. A case study is presented which uses a modular design with an integrated intelligent agent and knowledge base. It is argued that by using this type of approach, the real value of intelligent CBL to provide individual formative advice to students in a simulated experience can be realized. PMID- 11403586 TI - Similarities not differences: an exploration of the impact of change upon a group of nursing lecturers within a university setting. AB - The concepts of collaboration and partnership currently have extensive impact upon health care providers and higher education institutions. One of the challenges is to develop networks which will foster partnerships able to react, and contribute, to an ever-evolving educational culture. These themes are illustrated by using the example of one school of nursing and midwifery, and the collective experience of a number of its academic staff. By focusing on distinct features of collaboration (strategic planning, origins of change, group dynamics and building a community), the authors seek to explore the impact of an educational culture in an attempt to provide meaning to their recent experiences. In so doing, group identity is explored and the prospect for creating partnerships across disciplines ('similarities rather than differences') is considered. PMID- 11403587 TI - An approach to consensus building using the Delphi technique: developing a learning resource in mental health. AB - Health service research is best served through the collaboration of researchers and practitioners. This study utilizes a multi-method approach in developing a learning resource in clinical risk management in mental health. Phase one of the project is reported here; using a Delphi Technique to build the learning resource. Using the Delphi enabled a group of 'experts' to explore a complex issue and move towards consensus. In addition, the Delphi technique enabled the researchers to repeatedly access a dispersed group of 'experts' in an economic way. A three round Delphi was undertaken and results were analyzed (medians, means, standard deviations and thematic content analysis) to facilitate consensus building from a range of perspectives. The analytical model chosen is discussed in the context of prior use of the Delphi as a consensus building technique. The researchers conclude that analysis using medians and means to measure the level of agreement/disagreement with the content and structure of the learning pack, standard deviations to measure 'tightening' of the group view and Thematic Content Analysis of qualitative data was an effective approach. They argue that in the education and health fields, group views are particularly important and that this is a central tenet of the Delphi. PMID- 11403588 TI - An evaluation of a standards based portfolio. AB - In undergraduate and postgraduate nursing studies, the essay type assignment is commonly used to assess student learning. The literature suggests that of the alternatives for the assessment of learning, the use of portfolios may be an effective tool. A portfolio is generally conceptualized as a collection of the students best work or a collection of work that evidences incremental development. For a particular unit of learning, 'Nursing Business', second year undergraduate students in the Bachelor of Nursing programme were required to complete a portfolio. The assessment directed the students to meet specific criteria which in turn reflected the learning outcomes. Therefore the portfolio in this study is more appropriately described as a standards based portfolio (SBP). A SBP is a series of student work that seeks to address pre-determined learning outcomes. This study is an evaluation of student perceptions of a SPB and of interest to the study was the relationship between theory and practice, the availability of resources to complete the assignment and the contribution the portfolio made to the process of learning. PMID- 11403591 TI - Self-assembling, cystine-derived, fused nanotubes based on spirane architecture: design, synthesis, and crystal structure of cystinospiranes. AB - A novel family of cystine-based spirobicyclic peptides (cystinospiranes) has been synthesized by a single-step procedure involving condensation of pentaerythritol derived tetrachloride with either the simple L-cystine dimethyl ester or its C,C' extended bispeptides leading to a variety of 19-membered spirobicyclic peptides or its N,N'-extended bispeptides affording the ring-expanded 25-membered cystinospiranes. The design is flexible with respect to the ring size that can be adjusted depending upon the length of the N,N'-extended cystine bispeptide, and the choice of an amino acid, as illustrated here with the preparation of a large number of cystinospiranes containing a wide variety of amino acids. X-ray crystal structure of the parent spirane (5a) revealed nanotube formation by vertical stacking of relatively flat spirobicyclic molecules through contiguous NH- - O==C hydrogen bonding. The fused pair of parallel nanotubes is open-ended, hollow, and extends to infinity. Crystallographic parameters are the following: C(33)H(52)N(4)O(16)S(4), space group C2, a = 42.181(3) A, b = 5.1165(7) A, c = 11.8687(9) A, beta = 106.23(1) degrees. PMID- 11403592 TI - Different mechanisms of oxidative stress and neurotoxicity for Alzheimer's A beta(1--42) and A beta(25--35). AB - Oxidative stress induced by amyloid beta-peptide (A beta) has been implicated in the neurodegeneration observed in Alzheimer's disease (AD) brain. However, the mechanism by which the predominant form of A beta found in AD brains, A beta(1- 42), causes oxidative stress and neurotoxicity remains unknown. Numerous laboratories have used the smaller 11-amino acid fragment of the full-length peptide, A beta(25--35), as a convenient alternative in AD investigations since the smaller peptide mimics several of the toxicological and oxidative stress properties of the native full-length peptide. Our observation that the truncated peptide is more rapidly toxic and causes more oxidative damage than the parent A beta(1--42) led us to investigate the cause for this enhanced toxicity of A beta(25--35) in order to gain insight into the mechanism of action of these peptides. These studies reveal that two different mechanisms may be operative in the two peptides; however, the single methionine residue in the peptides appears to play a crucial role in both mechanisms. That methionine is C-terminal in A beta(25--35) seems to be the cause for its exaggerated effects. When the next amino acid in the sequence of A beta(1--42) (valine) is appended to A beta(25- 35), the resultant peptide, A beta(25--36), in which methionine is no longer C terminal, is neither toxic to cultured neurons nor does it cause oxidative damage. Additionally, oxidizing the sulfur of methionine to a sulfoxide abrogates the damaging effects of both A beta(25--35) and A beta(1--42). The putative mechanistic role of methionine in the observed properties of A beta peptides is discussed in the context of the obtained results as is the role of A beta(1--42) induced oxidative stress in the neurodegeneration found in AD brain. PMID- 11403593 TI - A novel single-molecule study to determine protein--protein association constants. AB - Atomic force microscopy (AFM) is traditionally used as an imaging technique to gain qualitative information for a biological system. We have successfully used the imaging capabilities of the AFM to determine protein-protein association constants. We have developed a method to measure the molecular weight of a protein based on its volume determined from AFM images. Our volume determination method allows for rapid, accurate analysis of large protein populations. On the basis of the measured volume, the fraction of monomers as dimers was determined for the DNA helicase UvrD, and the dissociation constant (K(d)) for the helicase was calculated. We determined a K(d) for UvrD of 1.4 microM, which is in good agreement with published K(d) data obtained from analytical ultracentrifugation (AUC) studies. Our method provides a rapid method for determining protein-protein association constants. PMID- 11403594 TI - Construction of regulated nanospace around a porphyrin core. AB - A series of 1,3,5-phenylene-based rigid dendritic porphyrins were synthesized by Suzuki coupling between a porphyrin core and dendron units. The intramolecular energy transfer was studied by absorption and fluorescence spectroscopies. The encapsulation of the porphyrin core within the 1,3,5-phenylene dendron units was found to provide highly efficient energy transfer from the dendron units to the porphyrin core. The dendritic wedge structure affected the energy transfer efficiency. The 1,3,5-phenylene-based rigid dendron units act as highly efficient light-harvesting antennae. These dendritic porphyrins have also been examined as C(60) hosts and substrate-selective oxidation catalysts. The attachment of the second generation of 1,3,5-phenylene-based dendron units with the porphyrin core enabled a stable inclusion of C(60) in toluene. Furthermore, the size and shape of the nanospace in the rigid dendritic porphyrins were found to affect the selectivity of substrates in the catalytic olefin oxidations. PMID- 11403595 TI - Binding site elucidation of hydantoin-based antagonists of LFA-1 using multidisciplinary technologies: evidence for the allosteric inhibition of a protein--protein interaction. AB - The binding site on the lymphocyte function-associated antigen-1 (LFA-1) of a class of hydantoin-based antagonists of leukocyte cell adhesion has been identified. This site resides in the inserted-domain (I-domain) of the CD11a chain at a location that is distal to residues known to be required for interactions with the intercellular adhesion molecules. This finding supports the hypothesis that the molecules are antagonizing cell adhesion via an allosteric modification of LFA-1. The binding site was identified using an integrated immunochemical, chemical, and molecular modeling approach. Antibodies that map to epitopes on the I-domain were blocked from binding to the purified protein by the hydantoins, indicating that the hydantoin-binding site resides on the I-domain. Photoaffinity labeling of the I-domain followed by LC/MS and LC/MS/MS analysis of the enzymatic digest identified proline 281 as the primary amino acid residue covalently attached to the photoprobe. Distance constraints derived from this study coupled with known SAR considerations allowed for the construction of a molecular model of the I-domain/inhibitor complex. The atomic details of the protein/antagonist interaction were accurately predicted by this model, as subsequently confirmed by the X-ray crystal structure of the complex. PMID- 11403596 TI - Synthesis, aggregation, and adsorption phenomena of shape-persistent macrocycles with extraannular polyalkyl substituents. AB - The synthesis of shape-persistent macrocycles based on the phenyl-ethynyl backbone containing various extraannular alkyl side chains is described. Although compound solubility increases with increasing size of the side groups, decreasing the solvent polarity induces aggregation of the rings by nonspecific interactions. This was investigated by proton NMR spectroscopy. The magnitude of aggregation can be varied by using solvent mixtures of different hexane content, supporting the model of a solvophobic effect. From 1,2,4-trichlorobenzene solution the macrocycle 1c adsorbs at the surface of highly oriented pyrolitic graphite (HOPG). The two-dimensional order of the structure was investigated by scanning tunneling microscopy (STM) revealing the formation of a two-dimensional lattice of p1(2)mm symmetry with lattice parameters A = 3.6 nm, B = 5.7 nm, and Gamma = 74 degrees. PMID- 11403597 TI - Helix unfolding in unsolvated peptides. AB - The conformations of unsolvated Ac-K(AGG)(5)+H(+) and Ac-(AGG)(5)K+H(+) peptides (Ac = acetyl, A = alanine, G = glycine, and K = lysine) have been examined by ion mobility measurements over a wide temperature range (150-410 K). The Ac K(AGG)(5)+H(+) peptide remains a globule (a compact, roughly spherical structure) over the entire temperature range, while both an alpha-helix and a globule are found for Ac-(AGG)(5)K+H(+) at low temperature. As the temperature is raised the alpha-helix unfolds. Rate constants for loss of the helix (on a millisecond time scale) have been determined as a function of temperature and yield an Arrhenius activation energy and preexponential factor of 38.2 +/- 1.0 kJ mol(-1) and 6.5 +/ 3.7 x 10(9) s(-1), respectively. The alpha-helix apparently does not unfold directly into the globule, but first converts into a long-lived intermediate which survives to a significantly higher temperature before converting. According to molecular dynamics simulations, there is a partially untwisted helical conformation that has both a low energy and a well-defined geometry. This special structure lies between the helix and globule and may be the long-lived intermediate. PMID- 11403598 TI - High-resolution structure and conformational dynamics of rigid, cofacially aligned porphyrin-bridge-quinone systems as determined by NMR spectroscopy and ab initio simulated annealing calculations. AB - The high-resolution solution structure and dynamics of a cofacially aligned porphyrin--phenylene--quinone compound have been determined using (1)H NMR spectroscopy and simulated annealing calculations. Members of this class of pi stacked assemblies feature a 1,8-naphthyl pillaring motif that enforces sub van der Waals interplanar separations between juxtaposed porphyryl, aromatic bridge, and quinonyl components of the donor--spacer--acceptor compound; this structural motif gives rise to a comprehensive set of structurally significant NOE signatures that can be used as constraints in quantitative structural calculations. Examination of such data using ab initio simulated annealing analytical methods shows that 5-[8'-(4' '-[8' "-(2' " ',5' "-benzoquinonyl)-1' " naphthyl]-1' '-phenyl)-1'-naphthyl]-10,20-diphenylporphyrin displays an unusual degree of conformational homogeneity in the condensed phase, and represents a rare example where such an analysis determines unequivocally a single unique structure in solution. PMID- 11403599 TI - A quantum chemical survey of metalloporphyrin-nitrosyl linkage isomers: insights into the observation of multiple FeNO conformations in a recent crystallographic determination of nitrophorin 4. AB - Using density functional theory-based geometry optimizations, we have searched for eta(1)-NO, eta(1)-ON (isonitrosyl), and eta(2)-NO (side-on bound NO) linkage isomers of a number of metalloporphyrin-NO complexes, M(Por)(NO)(L), where Por = porphinato dianion, M = Mn(II), Fe(II), Fe(III), Ru(II), Ru(III), Co(II), and Rh(II), and L = no ligand, SMe, Ph, and imidazole. The eta(1)-NO isomer had the lowest energy in all cases, and the isonitrosyl isomer was also located as a higher energy potential energy minimum in a number of cases. The eta(2)-NO isomer was only located as a minimum for Mn(II) (L = no ligand), Fe(III) (L = no ligand), and Ru(III) (L = Ph, imidazole, pyrdine), suggesting that an [MNO](6) electron count is important for stabilization of the eta(2) mode of ligation. However, in the presence of axial ligands L, the side-on isomers of [FeNO](6) complexes were not stable and opened up to an unusual geometry where the FeN(O) and NO vectors were tilted in opposite directions relative to the heme normal. Exactly such a geometry, as well as a "normal" upright geometry, has been observed in a recent crystallographic determination of nitrophorin 4 (Nature Struct. Biol. 2000, 7, 551), a salivary protein from the blood-sucking insect Rhodnius prolixus. Together, the calculated and experimental result illustrate the extreme softness of the FeNO potential energy surface toward various forms of tilting and bending deformations. PMID- 11403600 TI - Solvent effects on charge transfer bands of nitrogen-centered intervalence compounds. AB - Electron transfer parameters are extracted from the optical spectra of intervalence bis(hydrazine) radical cations. Compounds with 2-tert-butyl-3-phenyl 2,3-diazabicyclo[2.2.2]octyl-containing charge-bearing units that are doubly linked by 4-sigma-bond and by 6-sigma-bond saturated bridges are compared with ones having tert-butylisopropyl- and diphenyl-substituted charge bearing units and others having the aromatic units functioning as the bridge. Solvent effect studies show that the optical transition energy (E(op)) does not behave as dielectric continuum theory predicts but that solvent reorganization energy may be usefully separated from the vibrational reorganization energy by including linear terms in both the Pekar factor (gamma) and the Gutmann donor number (DN) in correlating the solvent effect. Solvation of the bridge for these compounds is too large to ignore, which makes dielectric continuum theory fail to properly predict solvent effects on either E(op) or the free energy for comproportionation. PMID- 11403601 TI - Selective intercalation of charge neutral intercalators into GG and CG steps: implication of HOMO-LUMO interaction for sequence-selective drug intercalation into DNA. AB - We have synthesized naphthopyranone epoxide 4 from D-isoascorbic acid together with its three diastereoisomers. DNA alkylation of ODNs containing 5'XGT3' and 5'TGY3' by 4 (11R, 13R), where X and Y are any nucleotide bases, occurred at all G residues except at G of the 5'TGC3' sequence. In contrast, the three other diastereoisomers of 4 showed only weak G alkylation activity. Differential (1)H NMR NOE of the 4-G adduct confirmed the G-N7 alkylation at the epoxide carbon of 4 with concomitant S(N)2 ring opening of the epoxide. Quantitative HPLC analysis of G alkylation efficiency for 4 showed the order of G alkylation susceptibility as TGGT approximately CGT >> TGA > AGT > TGT >> TGC. The order was fully consistent with those reported for aflatoxin B(1) oxide and kapurimycin A(3), suggesting that the sequence selectivity observed for these DNA alkylating agents is not structure dependent but most likely due to the intrinsic property of DNA sequences. We found that the order of G alkylation susceptibility obtained for 4 completely matched the calculated HOMO energy level of G-containing sequences. These results underscore that 4 is a unique molecular probe for ranking the HOMO level of G-containing sequences by well-known G alkylation chemistry and suggests that the intercalation of charge neutral intercalators is a HOMO-controlled process. PMID- 11403602 TI - Enhanced cage effects in supercritical fluid solvents. the behavior of diffusive and geminate caged-pairs in supercritical carbon dioxide. AB - The behavior of geminate and diffusive radical caged-pairs arising from the photolysis of dicumyl ketone in conventional and supercritical carbon dioxide (SC -CO(2)) solvents has been examined. The results suggest that locally enhanced solvent density about a solute (solvent/solute clustering) can lead to an enhanced cage effect near the critical pressure in supercritical fluid solvents. This enhanced cage effect is similar in magnitude for both diffusive and geminate caged-pairs. PMID- 11403603 TI - Dual-mode EPR study of Mn(III) salen and the Mn(III) salen-catalyzed epoxidation of cis-beta-methylstyrene. AB - Dual-mode electron paramagnetic resonance (EPR), in which an oscillating magnetic field is alternately applied parallel or perpendicular to the static magnetic field, is a valuable technique for studying both half-integer and integer electron spin systems and is particularly useful for studying transition metals involved in redox chemistry. We have applied this technique to the characterization of the Mn(III) salen (salen = N,N'-ethylene bis(salicylideneaminato)) complex [(R,R)-(-)-N,N'-bis(3,5-di-tert butylsalicylidene)-1,2-cyclohexanediaminomanganese(III)], with an S = 2 integer electron spin system. Furthermore, we have used dual-mode EPR to study the Mn salen complex during the Mn(III) salen-catalyzed epoxidation of cis-beta methylstyrene. Our study shows that the additives N-methylmorpholine N-oxide (NMO) and 4-phenylpyridine-N-oxide (4-PPNO), which are used to improve epoxidation yields and enantioselection, bind to the Mn(III) center prior to the epoxidation reaction, as evidenced by the alteration of the Mn(III) parallel mode EPR signal. With these additives as ligands, the axial zero-field splitting values and (55)Mn hyperfine splitting of the parallel mode signal are indicative of an axially elongated octahedral geometry about the Mn(III) center. Since the dual-mode EPR technique allows the observation of both integer and half-integer electron spin systems, Mn oxidation states of II, III, IV, and potentially V can be observed in the same sample as well as any radical intermediates or Mn(III,IV) dinuclear clusters formed during the Mn(III) salen-catalyzed epoxidation reaction. Indeed, our study revealed the formation of a Mn(III,IV) dinuclear cluster in direct correlation with expoxide formation. In addition to showing the possible reaction intermediates, dual-mode EPR offers insight into the mechanism of catalyst degradation and formation of unwanted byproducts. The dual-mode technique may therefore prove valuable for elucidating the mechanism of Mn(III) salen catalyzed reactions and ultimately for designing optimum catalytic conditions (solvents, oxidants, and additives such as NMO or 4-PPNO). PMID- 11403604 TI - Direct evidence for a geometrically constrained "entatic state" effect on copper(II/I) electron-transfer kinetics as manifested in metastable intermediates. AB - The absolute magnitude of an "entatic" (constrained) state effect has never been quantitatively demonstrated. In the current study, we have examined the electron transfer kinetics for five closely related copper(II/I) complexes formed with all possible diastereomers of [14]aneS(4) (1,4,8,11-tetrathiacyclotetradecane) in which both ethylene bridges have been replaced by cis- or trans-1,2-cyclohexane. The crystal structures of all five Cu(II) complexes and a representative Cu(I) complex have been established by X-ray diffraction. For each complex, the cross reaction rate constants have been determined with six different oxidants and reductants in aqueous solution at 25 degrees C, mu = 0.10 M. The value of the electron self-exchange rate constant (k(11)) has then been calculated from each cross reaction rate constant using the Marcus cross relation. All five Cu(II/I) systems show evidence of a dual-pathway square scheme mechanism for which the two individual k(11) values have been evaluated. In combination with similar values previously determined for the parent complex, Cu(II/I)([14]aneS(4)), and corresponding complexes with the two related monocyclohexanediyl derivatives, we now have evaluated a total of 16 self-exchange rate constants which span nearly 6 orders of magnitude for these 8 closely related Cu(II/I) systems. Application of the stability constants for the formation of the corresponding 16 metastable intermediates--as previously determined by rapid-scan cyclic voltammetry--makes it possible to calculate the specific electron self-exchange rate constants representing the reaction of each of the strained intermediate species exchanging electrons with their stable redox partners--the first time that calculations of this type have been possible. All but three of these 16 specific self-exchange rate constants fall within--or very close to--the range of 10(5)-10(6) M(-1) s( 1), values which are characteristic of the most labile Cu(II/I) systems previously reported, including the blue copper proteins. The results of the current investigation provide the first unequivocal demonstration of the efficacy of the entatic state concept as applied to Cu(II/I) systems. PMID- 11403605 TI - Getting a library of activities from a single compound: tunability and very large shifts in acidity constants induced by sol--gel entrapped micelles. AB - Acidity constants (in terms of K(i) values) can be fine-tuned, their values can be pushed to extremes, and their location can be placed either at the basic or acidic range of the pH scale, by tailoring intra-cage properties of SiO(2) sol- gel materials with surfactants. With the entrapped "solid-state pseudomicelles", an 8-orders magnitude K(i) shift--from the high-basic pH region to the acidic one -was obtained for acid fuchsin (AF); and tunability of the pK(i) values of the solvatochromic dye E(T)(30) and of crystal violet over much of the pH scale was achieved. This tailoring of various activities from the same dopant was achieved either by utilizing surfactants of different nature or by utilizing mixtures of anionic/cationic surfactants. The extraction of a wide range of reactivities from a single compound and the ability to fine-tune it is a powerful concept with potential extensions beyond acid/base reactions. PMID- 11403606 TI - Novel microporous europium and terbium silicates. AB - The synthesis and structural characterization of the first examples of microporous europium(III) and terbium(III) silicates (Na(4)K(2)X(2)Si(16)O(38) x 10H(2)O, X = Eu, Tb) are reported. The structure of these solids was solved by powder X-ray diffraction ab initio (direct) methods and further characterized by chemical analysis (EDS), thermogravimetric analysis (TGA), scanning electron microscopy (SEM), (23)Na and (29)Si magic-angle spinning (MAS) NMR, and luminescence spectroscopy. Both materials display interesting photoluminescence properties and present potential for applications in optoelectronics. This work illustrates the possibility of combining in a given silicate microporosity and optical activity. PMID- 11403607 TI - Synthesis of "solid solution" and "core-shell" type cobalt--platinum magnetic nanoparticles via transmetalation reactions. AB - In this article, we report the synthesis of "solid solution" and "core-shell" types of well-defined Co--Pt nanoalloys smaller than 10 nm. The formation of these alloys is driven by redox transmetalation reactions between the reagents without the need for any additional reductants. Also the reaction proceeds selectively as long as the redox potential between the two metals is favorable. The reaction between Co(2)(CO)(8) and Pt(hfac)(2) (hfac = hexafluoroacetylacetonate) results in the formation of "solid solution" type alloys such as CoPt(3) nanoparticles. On the other hand, the reaction of Co nanoparticles with Pt(hfac)(2) in solution results in "Co(core)Pt(shell)" type nanoalloys. Nanoparticles synthesized by both reactions are moderately monodispersed (sigma < 10%) without any further size selection processes. The composition of the alloys can also be tuned by adjusting the ratio of reactants. The magnetic and structural properties of the obtained nanoparticles and reaction byproducts are characterized by TEM, SQUID, UV/vis, IR, EDAX, and XRD. PMID- 11403608 TI - Improved resolution in proton NMR spectroscopy of powdered solids. AB - We present a new solid-state nuclear magnetic resonance experiment that yields, under CRAMPS decoupling conditions, a significant reduction in proton line widths for powdered organic solids. This experiment which relies on a constant-time acquisition of the proton transverse magnetization, removes the contribution of nonrefocusable broadening from the proton line widths. Although this new technique suffers from relatively low sensitivity, we demonstrate in this paper its feasibility on two model samples, L-alanine and the dipeptide Ala-Asp. In both cases a factor of between 2 and 3 in line width reduction is obtained for most of the proton resonances. PMID- 11403609 TI - Trajectory studies of S(N)2 nucleophilic substitution. 8. Central barrier dynamics for gas phase Cl(-) + CH(3)Cl. AB - Quasiclassical direct dynamics trajectories, calculated at the MP2/6-31G level of theory, are used to study the central barrier dynamics for the C1(-) + CH(3)Cl S(N)2 reaction. Extensive recrossings of the central barrier are observed in the trajectories. The dynamics of the Cl(-)-CH(3)Cl complex is non-RRKM and transition state theory (TST) is predicted to be an inaccurate model for calculating the Cl(-) + CH(3)Cl S(N)2 rate constant. Direct dynamics trajectories also show that Cl(-) + CH(3)Cl trajectories, which collide backside along the S(N)2 reaction path, do not form the Cl(-)-CH(3)Cl complex. This arises from weak coupling between the Cl(-)-CH(3)Cl intermolecular and CH(3)Cl intramolecular modes. The trajectory results are very similar to those of a previous trajectory study, based on a HF/6-31G* analytic potential energy function, which gives a less accurate representation of the central barrier region of the Cl(-) + CH(3)Cl reaction than does the MP2/6-31G* level of theory used here. Experiments are suggested for investigating the non-RRKM and non-TST dynamics predicted by the trajectories. PMID- 11403610 TI - A quantitative description of the ground-state wave function of Cu(A) by X-ray absorption spectroscopy: comparison to plastocyanin and relevance to electron transfer. AB - To evaluate the importance of the electronic structure of Cu(A) to its electron transfer (ET) function, a quantitative description of the ground-state wave function of the mixed-valence (MV) binuclear Cu(A) center engineered into Pseudomonas aeruginosa azurin has been developed, using a combination of S K-edge and Cu L-edge X-ray absorption spectroscopies (XAS). Parallel descriptions have been developed for a binuclear thiolate-bridged MV reference model complex ([(L(i)(PrdacoS)Cu)(2)](+)) and a homovalent (II,II) analogue ([L(i)(Pr2tacnS)Cu)(2)](2+), where L(i)(PrdacoS) and L(i)(Pr2tacnS) are macrocyclic ligands with attached thiolates that bridge the Cu ions. Previous studies have qualitatively defined the ground-state wave function of Cu(A) in terms of ligand field effects on the orbital orientation and the presence of a metal--metal bond. The studies presented here provide further evidence for a direct Cu--Cu interaction and, importantly, experimentally quantify the covalency of the ground-state wave function. The experimental results are further supported by DFT calculations. The nature of the ground-state wave function of Cu(A) is compared to that of the well-defined blue copper site in plastocyanin, and the importance of this wave function to the lower reorganization energy and ET function of Cu(A) is discussed. This wave function incorporates anisotropic covalency into the intra- and intermolecular ET pathways in cytochrome c oxidase. Thus, the high covalency of the Cys--Cu bond allows a path through this ligand to become competitive with a shorter His path in the intramolecular ET from Cu(A) to heme a and is particularly important for activating the intermolecular ET path from heme c to Cu(A). PMID- 11403611 TI - Rubbing-induced molecular reorientation on an alignment surface of an aromatic polyimide containing cyanobiphenyl side chains. AB - Surface lamellar decoration (SLD), surface enhanced Raman scattering (SERS) and optical second harmonic generation (SHG) experiments have been utilized to study the molecular orientation and conformation changes at a rubbed polyimide alignment-layer surface. This aromatic polyimide containing pendent cyanobiphenyl mesogens was synthesized via a polycondensation of 2,2'-bis(3,4-dicarboxy phenyl)hexafluoropropane dianhydride (6FDA) with bis[omega-[4-(4 cyanophenyl)phenoxy]hexyl] 4,4'-diamino-2,2'-biphenyldicarboxylate (nCBBP, n = 6), abbreviated as 6FDA--6CBBP. Uniform alignment layers, possessing high pretilt angles ranging from 39 degrees to 43 degrees, have been achieved after mechanical rubbing of the polyimide thin film surface at room temperature and subsequent annealing. This is the first time that high pretilt angles have been detected to possess a negative angle (-theta(c)) with respect to the rubbing direction (i.e., opposite to the rubbing direction), considerably different from the conventional pretilt angle (theta(c)) observed along the rubbing direction. This observation is confirmed using magnetic null and SHG methods. Combined polyethylene (PE) SLD and atomic force microscopy experiments reveal that the azimuthal orientation distribution of the long axis of the edge-on PE lamellar crystals is oriented normal to the rubbing direction, indicating that the PE chains are aligned parallel to the rubbing direction. This SLD technique probes the anisotropic surface orientation of the outermost molecules of the rubbed polyimide layer. The SERS results show that prior to rubbing the surface, both the pendent cyanobiphenyls in the side chains and backbones possess nearly planar chain conformations at the polyimide surface. Mechanical rubbing causes not only tilting of the backbone moieties, such as imide-phenylene structure, but also significant conformational rearrangements of the pendent side chains at the surfaces. The molecular mechanism of this unusual alignment is due to the fact that the pendent cyanobiphenyls forms a uniformly tilted conformation on the rubbed surface, and the polar cyano groups point down toward the layer surface deduced from SHG phase measurements. This conformational rearrangement of the side chains results in the formation of fold-like bent structures on the surface, which directly leads to the long axis of cyanobiphenyls having the -theta(c) pretilt angle with respect to the rubbing direction. PMID- 11403612 TI - A theoretical study on the mechanism and diastereoselectivity of the Kulinkovich hydroxycyclopropanation reaction. AB - A detailed mechanism for the Kulinkovich hydroxycyclopropanation reaction has been explored with density functional theory calculations on the reactions between R(1)COOMe and Ti(OMe)(2)(CH(2)CHR(2)) (R(1) and R(2) are hydrogen and alkyl groups). Addition of ester to titanacyclopropane is found to be fast, exothermic, and irreversible. It has a preference for the alpha-addition manifold over the beta-addition manifold in which its cycloinsertion transition states suffer from the steric repulsion between the R(2) and ester. The following intramolecular methoxy migration step is also exothermic with reasonable activation energy. The cyclopropane-forming step is the rate-determining step, which affords the experimentally observed cis-R(1)/R(2) diastereoselectivity in the alpha-addition manifold by generating cis-R(1)/R(2) 1,2-disubstituted cyclopropanol when R(1) is primary alkyl groups. On the contrary, the unfavored beta-addition manifold offers the diastereoselectivity contradicting the experimental observations. The effects of R(1) and R(2) on the regio- and stereoselectivity are also discussed. PMID- 11403613 TI - Inversion versus retention of configuration for nucleophilic substitution at vinylic carbon. AB - A high-level computational study using CCSD, CCSD(T), and G2(+) levels of theory has shown that unactivated vinyl substrates such as vinyl chloride would afford gas phase, single-step halide exchange by a pure in-plane sigma-approach of the nucleophile to the backside of the C--Cl sigma bond. Geometry optimization by CCSD/6-31+G* and CCSD(T)/6-31+G* confirms the earlier findings of Glukhovtsev, Pross, and Radom that the S(N)2 reaction of Cl(-) with unactivated vinyl chloride in the gas phase occurs by a sigma attack. Complexation of vinyl chloride with Na(+) does not alter this in-plane sigma preference. However, moderately activated dihaloethylenes such as 1-chloro-1-fluoroethylene undergo gas-phase S(N)2 attack by the accepted pi-route where the nucleophile approaches perpendicular to the plane of the C==C. In the latter case a single-step pi pathway is preferred for the Cl(-) + H(2)C==CFCl reaction. This is the first definitive example at a high level of theory where a single-step pi nucleophilic vinylic substitution is preferred over a multistep mechanism in the gas phase. The activation barriers for these gas-phase single-step sigma- and pi-processes involving both naked anions and Na(+) complexes are, however, prohibitively high. Solvation and the presence of a counterion must play a dominant role in nucleophilic vinylic substitution reactions that proceed so readily in the condensed phase. In solution, nucleophilic vinylic substitution reactions involving electron-withdrawing groups on the carbon--carbon double bond (e.g., CN, -CHO, and -NO(2)) would almost certainly proceed via a free discrete carbanionic intermediate in accord with experiment. PMID- 11403614 TI - Matrix photochemistry of syn-(chlorocarbonyl)sulfenyl bromide, syn-ClC(O)SBr: precursor to the novel species anti-ClC(O)SBr, syn-BrC(O)SCl, and BrSCl. AB - The vapor of (chlorocarbonyl)sulfenyl bromide, ClC(O)SBr, was isolated in solid Ar, Kr, N(2), and Ar doped with 5% CO at 15 K, and the matrix was subsequently irradiated with broad-band UV--visible light (200 < or = lambda < or = 800 nm), the changes being followed by reference to the IR spectrum of the matrix. The initial spectrum showed the vapor of ClC(O)SBr to consist of more than 99% of the syn form (with the C==O bond syn with respect to the S--Br bond) in equilibrium with less than 1% of the anti conformer. Irradiation caused various changes to occur. First, conformational randomization took place, leading to a roughly equimolar mixture of the two rotamers, and so affording the first spectroscopic characterization of an anti-ClC(O)S-containing compound. Simultaneously, the novel constitutional isomer syn-BrC(O)SCl was also formed. Continued photolysis resulted in the decay of all these species while revealing a third reaction channel, leading to the elimination of CO and the formation of the new triatomic sulfur halide BrSCl. The assignment of the IR bands to the different products was made on the basis of the usual criteria, taking account (i) of the effects of the naturally occurring isotopic pairs (35)Cl/(37)Cl and (79)Br/(81)Br, (ii) of the vibrational properties of related molecules, and (iii) of the properties predicted for the relevant molecules by quantum chemical calculations. PMID- 11403615 TI - The "wired" laccase cathode: high current density electroreduction of O(2) to water at +0.7 V (NHE) at pH 5. PMID- 11403616 TI - Uracil radicals in the gas phase: specific generation and energetics. PMID- 11403617 TI - Determining stoichiometry in homomultimeric nucleic acid complexes using magnetic field induced residual dipolar couplings. PMID- 11403618 TI - tert-Butoxyl as a model for radicals in biological systems: caveat emptor. PMID- 11403619 TI - Highly substituted ter-cyclopentanes as receptors for lipid A. PMID- 11403620 TI - The binuclear iridium(II) hydride complex [(C(5)Me(5))Ir(mu-H)](2): a novel base for reversible deprotonation of acidic organic compounds and a unique catalyst for C--C bond cleavage of aromatic 1,2-diols and Michael additions. PMID- 11403621 TI - Generation and reaction of metal-containing carbonyl ylides: tandem [3 + 2] cycloaddition--carbene insertion leading to novel polycyclic compounds. PMID- 11403622 TI - C-C bond-forming reductive elimination of ketones, esters, and amides from isolated arylpalladium(II) enolates. PMID- 11403623 TI - Novel bis(oxazoline) pincer ligands: formation of mononuclear rhodium(II) complexes. PMID- 11403624 TI - The theoretical transition state structure of a model complex bears a striking resemblance to the active site structure of DMSO reductase. PMID- 11403625 TI - Remarkably broad substrate tolerance of malonyl-CoA synthetase, an enzyme capable of intracellular synthesis of polyketide precursors. PMID- 11403626 TI - The lesser "burden borne" by o-succinylbenzoate synthase: an "easy" reaction involving a carboxylate carbon acid. PMID- 11403628 TI - Attachment of gold nanoparticles to glassy carbon electrodes via a mercaptobenzene film. PMID- 11403627 TI - Direct synthesis of beta-mannans. A hexameric [-->3)-beta-D-Man-(1](3) subunit of the antigenic polysaccharides from Leptospira biflexa and the octameric (1-->2) linked beta-D-mannan of the Candida albicans phospholipomannan. X-ray crystal structure of a protected tetramer. PMID- 11403629 TI - A polymeric chain in which mu(2),eta(2):eta(2)-ethene symmetrically bridges tetra(trifluoroacetato) dirhodium molecules. PMID- 11403630 TI - Charge and spin ordering in monoclinic Nb(12)O(29). PMID- 11403631 TI - Interactions of anionic carboranylated porphyrins with DNA. PMID- 11403632 TI - Exceptionally low-temperature carbon--hydrogen/carbon--deuterium exchange reactions of organic and organometallic compounds catalyzed by the Cp*(PMe(3))IrH(ClCH(2)Cl)(+) cation. PMID- 11403633 TI - Relativistic DFT calculations of the paramagnetic intermediates of [NiFe] hydrogenase. Implications for the enzymatic mechanism. PMID- 11403634 TI - Catalytic enantioselective synthesis of prostaglandin E(1) methyl ester using a tandem 1,4-addition-aldol reaction to a cyclopenten-3,5-dione monoacetal. PMID- 11403635 TI - The first catalytic asymmetric aza-Henry reaction of nitronates with imines: a novel approach to optically active beta-nitro-alpha-amino acid- and alpha,beta diamino acid derivatives. PMID- 11403636 TI - Mechanism of hydrogen sorption in single-walled carbon nanotubes. PMID- 11403638 TI - C-H...X--R (X = Cl, Br, and I) hydrogen bonds drive the complexation properties of a nanoscale molecular basket. PMID- 11403637 TI - Selective recognition of an alkali halide contact ion-pair. PMID- 11403641 TI - Signal transduction in neutrophil chemotaxis. AB - This review discusses current knowledge on signal transduction pathways controlling chemotaxis of neutrophils and similar cells. Most neutrophil chemoattractants bind to seven-transmembrane-helix receptors. These receptors activate trimeric G proteins of the Gi class in neutrophils to initiate chemotaxis. Phospholipases Cbeta, phosphoinositide 3-kinase gamma, and PH domain containing proteins play various roles in signaling further downstream. The actin cytoskeleton is crucial for cell motility, and is controlled by Rho family GTP binding proteins. PIP 5-kinase, LIM kinase, myosin light chain kinase and phosphatase, or WASP-like proteins may be important links between Rho GTPases and actin during chemotaxis. Newly emerging ideas on the regulation of the "compass" of chemotaxing cells, which may involve Cdc42 and certain PH domain-containing proteins, are also presented. PMID- 11403642 TI - Characterization of the lipopolysaccharide and structure of the O-specific polysaccharide of the bacterium Pseudomonas syringae pv. atrofaciens IMV 948. AB - Lipopolysaccharide (LPS) was isolated from the phytopathogenic bacterium Pseudomonas syringae pv. atrofaciens IMV 948 by mild extraction of the microbial cells with saline, and the properties, composition, and structure of the LPS were studied. The LPS showed low toxicity in D- galactosamine-sensitized mice and low biological activity in plants. Structural components of LPS--lipid A, core oligosaccharide, and O-specific polysaccharide (OPS)--were obtained by mild acid degradation and characterized. The lipid A contained fatty acids 3-HO-C10:0, C12:0, 2-HO-C12:0, 3-HO-C12:0, C16:0, C16:1, C18:0, and C18:1, as well as components of the hydrophilic moiety: GlcN, ethanolamine, phosphate, and phosphoethanolamine. The LPS core contained components typical of pseudomonads: glucose, rhamnose (Rha), L-glycero-D-manno-heptose, GlcN, GalN, 2-keto-3-deoxy-D manno-octonic acid, alanine, and phosphate. The OPS consisted of L-Rha and D GlcNAc in the ratio 4 : 1 and was structurally heterogeneous. The main pentasaccharide repeating unit of the OPS has the following structure: [structure see text]. Immunochemical studies showed that P. syringae pv. atrofaciens IMV 948 is serologically separate from other P. syringae strains, including those that have structurally similar OPS. PMID- 11403643 TI - Isolation of milk-clotting enzyme from transgenic sheep milk and its comparison with calf chymosin. AB - Technology for preparation of chymosin from milk of transgenic sheep has been elaborated. Purification of the preparation by ion-exchange chromatography on aminosilochrom and biospecific chromatography on bacitracin-Sepharose yielded homogeneous active enzyme. Hydrolysis of protein substrates (hemoglobin, BSA, and sodium caseinate) by the transgenic sheep chymosin and stability of the enzyme at various values of pH were studied. Judging by the amino acid composition, the N terminal sequence involving six amino acid residues, molecular mass, stability at various pH values, and the catalytic activity against the protein substrates, the transgenic sheep chymosin is identical to calf chymosin. PMID- 11403644 TI - Soybean bowman--birk inhibitor conjugates with clinical dextran. synthesis and antiproteolytic activity. AB - Conjugates of the classical soybean Bowman-Birk inhibitor (BBI) with clinical dextran were synthesized. Clinical dextran was preliminarily oxidized with periodate to dialdehydedextran (DAD). The effect of the degree of oxidation of DAD on coupling of the inhibitor was evaluated. The binding of the protein was shown to increase with increasing degree of DAD oxidation (5, 10, 20%). Total coupling of the inhibitor occurred when the degree of oxidation of the dextran was 20%. The BBI-DAD (20%) conjugate contained 13% protein with BBI/DAD molar ratio 1 : 1. The conjugates retained the ability to inhibit trypsin (Ki = 0.2-0.3 nM) and alpha-chymotrypsin (Ki = 15-30 nM). Thus, the coupling of BBI with the polymeric carrier caused practically no decrease in the antiproteolytic activity of the inhibitor. PMID- 11403645 TI - Fluorescence decay time distribution analysis reveals two types of binding sites for 1,8-anilinonaphthalene sulfonate in native human oxyhemoglobin. AB - Binding of 1,8-anilinonaphthalene sulfonate (1,8-ANS) with native human oxyhemoglobin (Hb) in 50 mM potassium phosphate buffer (pH 7.4) was studied by steady-state fluorescence spectroscopy and by laser spectrofluorimetry with subnanosecond time resolution. The distribution of fluorescence decay times and parameters of two- and three-exponential deconvolution of the fluorescence kinetics of 1,8-ANS in Hb solution demonstrate that the emission at wavelengths lambdaem of 455-600 nm is not single-exponential and has components with mean decay times < 0.5, 3.1-5.5, and 12.4-15.1 nsec with the amplitudes depending on the emission wavelength. Analysis of time-resolved fluorescence spectra shows that the shortest-lived component should be assigned to 1,8-ANS molecules in the aqueous medium, whereas the two longer-lived components are assigned to two types of binding sites for 1,8-ANS in the Hb molecule characterized by different polarity and accessibility to water molecules. PMID- 11403646 TI - Sialyltransferase activity in normal and atherosclerotic human aorta intima. AB - Sialyltransferase activity has been determined in Golgi membrane fractions isolated from atherosclerotic and normal intima of human aorta by measuring the transfer of N-acetylneuraminic acid (NeuAc) from CMP-NeuAc to asialofetuin. The asialofetuin-sialyltransferase activity was found to be twofold higher in the atherosclerotic intima than in the normal intima. The mean value of the apparent Michaelis constant (Km) for the sialylating enzyme in both tissues did not differ and was 57 microM. In contrast, the maximal velocity (Vmax) was 2-fold higher for the atherosclerotic intima than for the normal intima. These results suggest that expression of asialofetuin-sialyltransferases of the aortal intima may be increased in atherosclerosis. PMID- 11403647 TI - Interaction of loach DNA polymerase delta with DNA duplexes with single-strand gaps. AB - The interaction of DNA polymerase delta purified from eggs of the teleost fish Misgurnus fossilis (loach) with DNA duplexes with single-strand gaps of 1-13 nucleotides was studied. In the absence of template-restricting DNA, the enzyme elongated primers on single-stranded DNA templates in a distributive manner. However, in the presence of the proximal 5;-terminus restricting the template, the enzyme activity significantly increased. In this case, the enzyme was capable of processive synthesis by filling gaps of 5-9 nucleotides in DNA duplexes. These data indicate that DNA polymerase delta can interact with both the 3;- and 5; termini located upstream and downstream from the gap. Analysis of the complexes formed by DNA polymerase delta and different DNA substrates by electrophoretic mobility shift assay confirmed the assumption that this enzyme can interact with the proximal 5;-terminus restricting the gap. DNA polymerase delta displayed much higher affinity in duplexes with gaps of approximately 10 nucleotides compared to the standard template-primer complexes. Maximal affinity was observed in experiments with DNA substrates containing unpaired 3;-tails in primers. The results of this study suggest that DNA polymerase delta exerts high activity in the cell nuclei during repair of DNA intermediates with single-strand gaps and unpaired 3;-termini. PMID- 11403648 TI - Interaction of sulfite with the noncatalytic and catalytic sites of chloroplast coupling factor cf1. AB - The interaction between sulfite, an efficient Mg2+-dependent F1-ATPase activator, and chloroplast CF1-ATPase was studied. The sulfite anion was shown to inhibit ADP and ATP binding to the noncatalytic sites of CF1. The stimulating activity of sulfite persists when all noncatalytic sites are nucleotide-occupied. Phosphate, a competing candidate for binding to CF1 catalytic sites, suppresses this activity. These results support the suggestion that the stimulation of Mg2+ dependent ATPase activity of CF1 is caused by sulfite binding to its catalytic sites. PMID- 11403649 TI - Effects of culture method and growth phase on free lipid composition of Yersinia pseudotuberculosis. AB - The influence of culture method (free-floating cells in liquid nutrient broth or bacteria attached to agar surface on solid agarized medium of the same formulation) and bacterial age on the composition of free lipids in Yersinia pseudotuberculosis (O:Ib serovar, strain KS 3058) grown in the cold (5 degrees C) has been investigated. The specific growth rate of the bacteria on solid medium was about threefold less than that in liquid medium. The qualitative composition of phospholipids and fatty acids only slightly depended on the bacterial culture method. At the same time, the colonially growing cultures contained somewhat more total lipids, they synthesized more phospholipids, in the linear growth phase they contained more lysophosphatides, and they had higher fatty acid unsaturation index and higher pathogenic potential than their "planktonic" counterparts grown in otherwise identical conditions. The bacterial growth phase influenced the amount of 3-hydroxytetradecanoic acid and, indirectly, that of lipopolysaccharide. The dynamics of changes in the amount of this acid with bacterial age was opposite in the surface and broth cultures. PMID- 11403650 TI - Stimulation of mu and delta-Opiate Receptors and Tolerance of Isolated Heart to Oxidative Stress: the Role of NO-Synthase. AB - Preliminary intravenous injection of delta-opiate receptor (OR) agonists DSLET (0.1 mg/kg) or DTLET (0.1 mg/kg) increased tolerance of isolated perfused myocardium to damage by oxidative stress simulated in vivo with FeSO4 + ascorbic acid. This manifested itself by a decreased level of creatine phosphokinase (CPK) in the perfusate flowing out of the heart during the oxidative exposure. The preliminary systemic injection of mu-agonists DAMGO (0.1 mg/kg) or DALDA (0.1 mg/kg) failed to affect the release of CPK from the myocardium. The cardioprotective effect of the delta-agonist DSLET was completely abolished by preliminary intravenous injection of the delta-OR antagonist ICI 174,864 (2.5 mg/kg). The intravenous injection of the NO-synthase inhibitor L-NAME (50 mg/kg) also completely abolished the cardioprotective effect of delta-OR stimulation. The preliminary injection of DSLET but not of DAMGO prevented an increase in the level of diene conjugates and a decrease in the activity of superoxide dismutase (SOD) in the isolated myocardium tissue. Thus, the in vivo stimulation of delta OR increased the tolerance of the heart to oxidative stress through activation of NO-synthase and SOD. PMID- 11403651 TI - Characterization of bovine atrial angiotensin-converting enzyme. AB - Bovine atrial angiotensin-converting enzyme (ACE) was purified to electrophoretic homogeneity. The purification procedure included ion-exchange chromatography on DEAE-Toyopearl 650M, affinity chromatography on lisinopril-agarose and gel filtration on Sephadex G-100. The bovine atrial ACE exhibited similar sensitivities to inhibition by lisinopril and captopril as lung ACE (the Ki values for the atrial and lung enzymes differed insignificantly). However, the kinetic parameters of hydrolysis of some synthetic tripeptide substrates (FA-Phe Gly-Gly, FA-Phe-Phe-Arg, Cbz-Phe-His-Leu, Hip-His-Leu) catalyzed by bovine atrial and lung ACE varied to a greater extent. The enzymes were also characterized by some differences in activation by chloride, nitrate, and sulfate anions. These data support the hypothesis of tissue specificity of ACEs. PMID- 11403652 TI - H+/2e- stoichiometry of the nadh:ubiquinone reductase reaction catalyzed by submitochondrial particles. AB - Mitochondrial NADH:ubiquinone-reductase (Complex I) catalyzes proton translocation into inside-out submitochondrial particles. Here we describe a method for determining the stoichiometric ratio H+/2e- (n) for the coupled reaction of NADH oxidation by the quinone acceptors. Comparison of the initial rates of NADH oxidation and alkalinization of the surrounding medium after addition of small amounts of NADH to coupled particles in the presence of Q1 gives the value of n = 4. Thermally induced deactivation of Complex I [1, 2] results in complete inhibition of the NADH oxidase reaction but only partial inhibition of the NADH:Q1-reductase reaction. N-Ethylmaleimide (NEM) prevents reactivation and thus completely blocks the thermally deactivated enzyme. The residual NADH:Q1-reductase activity of the deactivated, NEM-treated enzyme is shown to be coupled with the transmembraneous proton translocation (n = 4). Thus, thermally induced deactivation of Complex I as well as specific inhibitors of the endogenous ubiquinone reduction (rotenone, piericidin A) do not inhibit the proton translocating activity of the enzyme. PMID- 11403653 TI - Acylation of Bowman-Birk soybean proteinase inhibitor by unsaturated fatty acid derivatives. AB - A procedure was developed for acylation of Bowman-Birk soybean proteinase inhibitor (BBI) by N-hydroxysuccinimide esters of oleic, linoleic, and alpha linolenic acids in a dimethyl sulfoxide-dioxane-pyridine mixture. BBI derivatives containing two acylated amino groups were prepared with high yield. The use of the reversible modifier citraconic anhydride in the first stage of synthesis permitted the synthesis of hydrophobized BBI derivatives retaining high antitrypsin and antichymotrypsin activities. It was found that the insertion of two long-chain moieties in the BBI molecule decreases its thermostability. PMID- 11403654 TI - Kinetics of denaturation of rabbit skeletal muscle glycogen phosphorylase b by guanidine hydrochloride. AB - The kinetics of denaturation and aggregation of rabbit muscle glycogen phosphorylase b in the presence of guanidine hydrochloride (GuHCl) have been studied. The curve of inactivation of phosphorylase b in time includes a region of the fast decline in the enzymatic activity, an intermediate plateau, and a part with subsequent decrease in the enzymatic activity. The fact that the shape of the inactivation curves is dependent on the enzyme concentration testifies to the dissociative mechanism of inactivation. The dissociation of phosphorylase b dimers into monomers in the presence of GuHCl is supported by sedimentation data. The rate of phosphorylase b aggregation in the presence of GuHCl rises as the denaturant concentration increases to 1.12 M; at higher concentration of GuHCl, suppression of aggregation occurs. At rather low concentration of the protein (0.25 mg/ml), the terminal phase of aggregation follows the kinetics of a monomolecular reaction (the reaction rate constant is equal to 0.082 min(-1); 1 M GuHCl, 25 degrees C). At higher concentration of phosphorylase b (0.75 mg/ml), aggregation proceeds as a trimolecular reaction. PMID- 11403655 TI - Resistance of dextran-modified hyaluronidase to inhibition by heparin. AB - Properties of native and aldehyde dextran-modified hyaluronidase (with surface amino group modification about 98%) were investigated. Optimal endoglycosidase activity of the native enzyme was observed at 0.15 M NaCl and pH 5.5 and electrostatic interactions influenced the enzyme activity. The inhibitory effect of heparin on hyaluronidase activity slightly differed at pH 5.5 (1.5-fold inhibition) and 7.5 (1.2-fold inhibition). Ionic strength of the reaction medium only slightly influenced the effect of heparin. Modification of hyaluronidase with dextran increased hydrophobic interactions and steric hindrance. Conjugation with dextran increased the resistance of hyaluronidase activity to denaturing agents (urea, guanidinium hydrobromide) and extended the optimal conditions for maximal endoglycosidase activity (pH 4.5-6.5, the range of NaCl concentration from 0.1 to 0.3 M). The conjugation also reduced electrostatic effects on the active site of hyaluronidase and efficacy of heparin inhibition. At pH 7.5 the enzyme was almost insensitive to heparin. The resistance of dextran-modified hyaluronidase to heparin points to approaches for subsequent studies of the heparin-binding site of this enzyme and biomedical trial of the stabilized enzyme for the treatment of acute cardiovascular lesions. PMID- 11403656 TI - 8th Congress of the International Society for Rotary Pumps. September 2000, Aachen, Germany. Proceedings. PMID- 11403657 TI - Progress and future perspectives in mechanical circulatory support. AB - Progress in several types of artificial organs in the cardiovascular field has significantly contributed to advancements in cardiac surgery. Due to the progress of high technology in fields other than medicine, both cardiac surgery and artificial organs have shown rapid and remarkable advances. In recent years, several types of blood pumps have been developed that are widely used not only as the main pump of cardiopulmonary bypass but also for circulatory support of postcardiotomy cardiogenic shock. In this article, the progress and current status of percutaneous cardiopulmonary support systems and ventricular assist devices (VADs) are described. In addition, new centrifugal and axial pumps, which are compact and implantable clinical use devices, are introduced. I believe that by making developments toward the clinical application of artificial hearts or VADs, not only in Japan but also in cooperation with colleagues at various institutions throughout the world, we will be able to make some contributions to the progress in the field of cardiac surgery. In the 20th century, medical research showed remarkable advances, mainly in medical electronics and pathophysiology. However, in the next century, we have to focus on other research fields, namely artificial organs and gene technology. PMID- 11403658 TI - Indications for rotary blood pumps in clinical practice. AB - To understand the needs and requirements in the development of rotary blood pumps, we have to consider the current use and limitations of blood pumps in cardiac surgery. There are 5 different fields of indications for blood pumps in cardiac surgery. First, there is the possibility to use rotary blood pumps in the cardiopulmonary bypass system. Second, miniaturized rotary blood pumps can be used to support beating heart surgery. Third, we expect a growing use of rotary blood pumps in short-term ventricular support. The same applies to the field of chronic cardiac failure and long-term myocardial support. Finally, miniaturized rotary blood pumps can be used to support the ventricle during myocardial infarction. Rotary blood pumps are expected, based on their features, to progress in all these indications. PMID- 11403659 TI - A new intracardiac microaxial pump: first results of a multicenter study. AB - As coronary artery bypass grafting (CABG) surgery in the beating heart technique is progressing, new devices have been developed to overcome hemodynamic instabilities while tilting the heart for exposure of back wall vessels. A new device for in heart biventricular intracorporeal circulation was applied in 42 patients undergoing CABG surgery (Group 1). The control group consisted of 38 patients operated on using a conventional cardiopulmonary bypass setup (Group 2). The study protocol of the prospective, randomized multicenter study was approved by the local ethics committees. Patients were included following inclusion criteria and patient informed consent. Mean age, procedure time, mean arterial pressure (MAP), and hemolysis by means of plasma free hemoglobin (fHb) were assessed preoperatively, perioperatively, on postoperative Days 1 to 3, at discharge, and at a 3 month follow-up. The mean age was 62.1 (range 59-74) years (Group 1), 62.7 (range 48-72) years (Group 2); procedure time was 112 min +/- 31.9 min (Group 1), 137.4 min +/- 36.2 min (Group 2); and 2.3 +/- 0.6 (Group 1), 2.2 +/- 0.7 (Group 2), vessels were revascularized. The flow on pump was 3.7 (2.5/4.4) L/min (Group 1), 4.9 (3.6/6.2) L/min (Group 2) which resulted in a MAP of 69.8 (4.0/143) mm Hg (Group 1), 58.3 (5.3/94) mm Hg while assessing the vessels of the back wall. Hemolysis defined by fHB was lower than 20 mg/dl at all times pre- and postoperatively. Intraoperative maximum values were up to 100 mg/dl in 4 patients (2 in Group 1 and 2 in Group 2). Body mass index was 26.4 +/- 2.6 (Group 1), 27.9 +/- 3.2. New York Heart Association Class was II to III in both groups. There were no pump related life threatening or severe adverse events. Beating heart procedures with ICC can be reliably and safely achieved. As the device is easy to use, it may deserve a more widespread use in the future. PMID- 11403660 TI - First experiences with outpatient care of patients with implanted axial flow pumps. AB - As known from patients with pulsatile ventricular assist devices (VADs), early mobilization, physical exercise, and return to normal life are essential for optimal recovery. Recently, implantable rotary pumps became available for extended left ventricular support as bridges to transplantation. Modified procedures are essential for patient training and hospital discharge. The MicroMed-DeBakey VAD was implanted in 10 patients with end-stage heart disease. After recovery, regular ergometer training was performed with loads adapted to the patient's condition. Procedures for patient observation under outdoor conditions and a blood pressure measuring device for low pulse pressure conditions were developed. Improvement of physical condition was achieved in 8 patients. In the first 2 patients, exercise capacity was limited due to flow obstruction. In the following patients, an increase of workload on the ergometer up to 120 W was observed. Correlated with training, lactate/load relationship and heart rate decreased. Three patients were discharged from the hospital during support. The DeBakey-VAD system can support patients for extended time periods and is suitable for recovery and exercise. Under optimal patient and environmental conditions, discharge from the hospital can be obtained. PMID- 11403661 TI - Computational fluid dynamics as a development tool for rotary blood pumps. AB - Computational fluid dynamics (CFD) is beginning to significantly impact the development of biomedical devices, in particular rotary cardiac assist devices. The University of Pittsburgh's McGowan Center for Artificial Organ Development has extensively used CFD as the primary tool to analyze and design a novel axial flow blood pump having a magnetically suspended rotor. The blood-contacting surfaces of the pump were developed using a design strategy based on CFD that involved closely coupling a Navier-Stokes solver to a parameterized geometry modeler and advanced mesh movement techniques. CFD-based blood damage models for shear-induced hemolysis as well as surrogate functions describing thrombosis potential were employed to help guide design improvements. This CFD-based design approach resulted in the timely development of a pump subjected to multiple geometric refinements without building expensive physical prototypes for each design iteration. A physical prototype of the final improved pump was fabricated and experimentally analyzed using particle imaging flow visualization. The CFD predicted results correlated well with the experimental data including pressure flow (H-Q) performance and specific flow field features. It is estimated that the present CFD-based design approach shortened the overall design time frame from an order of years to months. PMID- 11403662 TI - Assessment of hemolysis related quantities in a microaxial blood pump by computational fluid dynamics. AB - A computational assessment or even quantification of shear induced hemolysis in the predesign phase of artificial organs (e.g., cardiac assist devices) would largely decrease efforts and costs of design and development. In this article, a general approach of hemolysis analysis by means of computational fluid dynamics (CFD) is discussed. A validated computational model of a microaxial blood pump is used for detailed analysis of shear stress distribution. Several methods are presented that allow for a qualitative assessment of shear stress distribution and related exposure times using a Lagrangian approach and mass distribution in combination with shear stress analysis. The results show that CFD offers a convenient tool for the general assessment of shear-induced hemolysis. The determination of critical regions and an estimation of the amount of blood subject to potential damage in relation to the total mass flow are shown to be feasible. However, awareness of limitations and potential flaws in CFD based hemolysis assessments is crucial. PMID- 11403663 TI - Computational fluid dynamics performance prediction for the hydrodynamic bearings of the ventrassist rotary blood pump. AB - Finite-volume computations are described for laminar flow in the hydrodynamic bearings supporting 2 different versions of the impeller of the VentrAssist rotary pump. Pressure boundary conditions are taken from prior computations of turbulent flow in the whole pump with frictionless sliding of the impeller on the inside of the pump body. By investigating various impeller positions, the true ride height is determined. Net lift and combined drag from all 8 bearings of the 4-bladed impeller are compared with predictions based on 2-D theory. The computations also reveal the extent of net force and moment acting to move the impeller away from its nominal axis of rotation. PMID- 11403664 TI - Development of the MEDOS/HIA DeltaStream extracorporeal rotary blood pump. AB - The DeltaStream blood pump has been developed for extracorporeal circulation with one focus on potential integration into simplified bypass systems (SBS). Its small size and an embedded electric motor are the basic pump properties. A variation of the impeller design has been performed to optimize hydraulic and hematologic characteristics. A simple impeller design was developed which allows flow and pressure generation for cardiopulmonary bypass applications. The option of a pulsatile flow mode for ventricular assist device applications also was demonstrated in vitro. Impeller washout holes were implemented to improve nonthrombogenicity. The pump was investigated for potential thermal hazards for blood caused by the integrated electric motor. It could be demonstrated that there is no thermal risk associated with this design. Durability tests were performed to assess the lifetime of the pump especially with regard to the incorporated polymeric seal. Seal lifetimes of up to 28 days were achieved using different blood substitutes. In animal tests using either the pump as a single device or in an SBS setup, biocompatibility, low hemolysis, and nonthrombogenicity were demonstrated. In summary, the DeltaStream pump shows great potential for different extracorporeal perfusion applications. Besides heart-lung machine and SBS applications, ventricular assist and extracorporeal membrane oxygenation up to several days also appear promising as potential applications. PMID- 11403665 TI - Control strategy for rotary blood pumps. AB - The control strategy for ventricular support with a centrifugal blood pump was examined in this study. The control parameter was the pump rpm that determines pump flow. Optimum control of pump rpm that reflects the body's demand is important for long-term, effective, and safe circulatory support. Moreover, continuous, reliable monitoring of ventricular function will help successfully wean the patients from the ventricular assist device (VAD). The control strategy in this study includes determination of the target pump rpm that can provide the flow required by the body, fine-rpm-tuning to minimize deleterious effects such as suction in the ventricle, and assessment of ventricular function for successful weaning from VADs. To determine the target pump rpm, we proposed to use the relation between the native heart rate and cardiac output, and the relation between the pump rpm and centrifugal pump output. For fine-tuning of the pump rpm, the motor current waveform was used. We computed the power spectral density of the motor current waveform and calculated the ratio of the fundamental to the higher order components. When this ratio was larger than approximately 0.2, we assumed there would be a suction effect in the ventricle. As for assessment of ventricular function, we used the amplitude of the motor current waveform. The control system implemented using a DSP functioned properly in the mock circulatory loop as well as in acute animal experiments. The motor current also showed a good correlation with the ventricular pressure in acute animal experiments. PMID- 11403666 TI - Change in vasoconstrictive function during prolonged nonpulsatile left heart bypass. AB - We investigated changes in vasoconstrictive function accompanying prolonged nonpulsatile left heart bypass (NLHB). After 2-week pulsatile left heart bypass (PLHB) in 11 goats, NLHB was conducted for another 4 weeks (Group N) in 6 goats. In the other 5 goats, PLHB was continued for another 4 weeks (Group P). Systemic vascular resistance at rest (rSVR) was measured on the last days of the second and sixth postoperative week (W2 and W6, respectively). Subsequently, phenylephrine was injected, and the maximum values (SVRmax) and the maximum increasing change in SVR (DeltaSVR) were measured. No significant difference was observed in rSVR between groups at W2 or W6. The SVRmax and the DeltaSVR at W2 were consistent in both groups. However, at W6, the SVRmax and the DeltaSVR of Group N were significantly lower than those of Group P. In conclusion, prolonged NLHB caused a significant decrease in the SVR response to phenylephrine, indicating a dimunition of vasoconstrictive function. PMID- 11403667 TI - A blood pressure sensor for long-term implantation. AB - An implantable flow-through blood pressure sensor prototype has been developed for use with an implantable left ventricular assist device (LVAD). This sensor incorporates a flat pressure-sensing diaphragm that is designed to be integral with the wall of a titanium tube that may be placed in the inlet or the outlet flow path of any LVAD. The interior tube flow geometry is transitioned from a round to a D-shape such that flow separation is eliminated. Bench testing of 3 sensors was performed to characterize the sensor. The worst-case results showed a maximum nonlinearity of 0.64 mm Hg, a maximum hysteresis of 0.87 mm Hg, and a maximum nonrepeatability of 0.87 mm Hg. Long-term drift studies of 2 sensors at 193 days and 112 days resulted in a projected annual drift rate of 1.4 and 2.0 mm Hg, respectively. The APEX pressure sensors were evaluated in 5 ventricular assist acute calf experiments in which the sensor outputs were compared with Millar pressure catheter sensors. Pressure output comparisons showed similar pressure tracings. No visible evidence of thrombus formation was found on the APEX sensor compared with thrombus formation found on the Millar catheter at the entrance to the flow path. Tests demonstrated that the blood pressure sensor can accurately measure blood pressure and indicate that it has long-term stability. PMID- 11403668 TI - The Heartmate II: design and development of a fully sealed axial flow left ventricular assist system. AB - Our group is developing the control and power transmission components required to implement a permanent and fully sealed left ventricular assist system (LVAS). Starting with the percutaneously powered HeartMate II blood pump, our development efforts are focused in the following areas: a complete redesign of the transcutaneous energy transmission system (TETS) to include a rectification network and autonomous voltage regulation within the secondary coil, a hermetically sealed electronics package containing a miniaturized implementation of the existing redundant drive and control electronics with several power-input options, an implanted rechargeable lithium ion battery pack capable of providing up to 1 h of untethered operation, implantable electrical connectors that allow components to be connected after placement in the body or to be replaced if needed, and a radio telemetry subsystem to transmit diagnostic information and to permit remote adjustment of selected parameters. PMID- 11403669 TI - The Heartmate III: design and in vivo studies of a maglev centrifugal left ventricular assist device. AB - A compact implantable centrifugal left ventricular assist device (LVAD) (HeartMate III) featuring a magnetically levitated impeller is under development. The goal of our ongoing work is to demonstrate feasibility, low hemolysis, and low thrombogenicity of the titanium pump in chronic bovine in vivo studies. The LVAD is based on so-called bearingless motor technology and combines pump rotor, drive, and magnetic bearing functions in a single unit. The impeller is rotated (theta z) and levitated with both active (X, Y) and passive (Z, theta x, theta y) suspension. Six prototype systems have been built featuring an implantable titanium pump (69 mm diameter, 30 mm height) with textured blood contacting surfaces and extracorporeal electronics. The pumps were implanted in 9 calves (< or = 100 kg at implant) that were anticoagulated with Coumadin (2.5 < or = INR < or = 4.0) throughout the studies. Six studies were electively terminated (at 27 61 days), 1 study was terminated after the development of severe pneumonia and lung atelectasis (at 27 days) another study was terminated after cardiac arrest (at 2 days) while a final study is ongoing (at approximately 100 days). Mean pump flows ranged from 2 to 7 L/min, except for brief periods of exercise at 6 to 9 L/min. Plasma free hemoglobin ranged from 4 to 10 mg/dl. All measured biochemical indicators of end organ function remained within normal range. The pumps have met performance requirements in all 9 implants with acceptable hemolysis and no mechanical failures. PMID- 11403670 TI - Intravascular micropump for augmented liver perfusion: first in vivo experience. AB - The aim of this study was to assess the in vivo performance of a new microaxial rotary blood pump developed for long-term intraportal implantation. The pump, measuring 7 mm in diameter, has a single stage impeller and is powered by a microelectric motor. The pump was implanted into the portal vein in 13 large white pigs under general anesthesia. All animals recovered after the portal pump implantation, and they were observed until the pump failed. The 2 longest running pumps performed for 40 and 36 h, respectively. Either thrombus formation or technical problems, especially in the bearings, were the main causes of pump failure during the experiment. No local or systemic adverse effects were observed during the portal pumping period. Full recovery of the animals following intraportal pump implantation was achieved. However, further technical improvements to the pump are required to maintain a longer performance in vivo. PMID- 11403671 TI - An ultradurable and compact rotary blood pump with a magnetically suspended impeller in the radial direction. AB - A magnetically suspended centrifugal blood pump has been developed with a self bearing motor for long-term ventricular assist systems. The rotor of the self bearing motor is not only actively suspended in the radial direction, but also is rotated by an electromagnetic field. The pump has a long lifetime because there are no mechanical parts such as seals and motor bearings. An outer rotor mechanism was adopted for the self-bearing motor. The stator was constructed in the central space of the motor. The rotor shaped thin ring was set at the circumferential space of the stator. Six vanes were extended from the upper surface of the rotor toward the center of the pump to construct an open-type impeller. The outer diameter and the height of the impeller are 63 mm and 34 mm, respectively. The magnetic bearing method and the servomotor mechanism were adopted to levitate and rotate the rotor. Radial movements of the rotor and rotation are controlled actively by using electromagnets in the stator. Axial movement and tilt of the rotor are restricted by passive stability to simplify the control. The radial gap between the rotor and the stator is 1 mm. A closed loop circuit filled with water was used to examine basic performance of the pump. Maximum flow rate and pressure head were 8 L/min and 200 mm Hg, respectively. Maximum amplitude of radial displacement of the impeller was 0.15 mm. The impeller could be suspended completely without touching the casing wall during the entire pumping process. Power consumption of the pump was only 9.5 W to produce a flow rate of 5 L/min against a pressure head of 100 mm Hg. We conclude that the pump has sufficient performance for the implantable ventricular assist system. PMID- 11403672 TI - Recent progress on transcutaneous energy transfer for total artificial heart systems. AB - For many years, transcutaneous energy transfer (TET) systems have been developed for energizing total artificial heart systems. Although such a basic system can be developed without too much design effort, optimization toward high power transfer efficiency forces the introduction of novel system topologies and design strategies. In addition, for medical applications, the thermal impact of a TET system on the biological tissue should be taken into account, resulting in limitations on usable coil geometries. This article presents a TET system that has been developed for a power transfer of 25 W over a distance of 1 cm with minimal dimensions of 1 x 6 x 4 cm for the external driver and 5 x 3 x 1 cm for the internal electronics. The coil geometries have a thickness of 2 mm and a diameter of 6 cm. An overall system efficiency of 80% was achieved for an internal load of 25 W. PMID- 11403673 TI - Blood pumps: technologies and markets in transformation. AB - The heart failure industry or sector of blood pump technologies comprised of scientists, medicine, and business has inadequate attention from the financial community due to the perception of unmet market potential and regulatory hurdles. There are more than 20 mechanical circulatory support technologies currently in some stage of development. The application is defined initially for emerging technologies as bridge-to-transplant whereas more mature devices have focused on bridge-to-recovery and alternative-to-transplant. Regulatory hurdles and financing resources have dictated this initial application strategy. Whether a technology has valves, bearings, is magnetically suspended, acts as a pressure cuff, and so on, the future will belong in large part to those technologies that are less expensive, have improved ergonomics, are simpler in surgical application, and can demonstrate efficacy levels that are an improvement over today's approved devices. Today's devices have provided a valuable platform for the emerging technologies. However, there has been little market expansion over these past years, relative to the potential market. The cardiologist is the patient's gatekeeper, and drugs appear to be the therapy of choice as options are considered. Technologies without a business model will have a difficult time attracting much needed funding, the result being protracted development times or project termination. MicroMed has modified its ventricular assist device (VAD) focus, with a broader look at treating the heart failure patient. MicroMed recently entered into an agreement with Chrysalis Biotechnology, Inc., Galveston, TX, covering a synthetic peptide technology (Chrysalin) that when injected into tissue has demonstrated an angiogenic effect without the concerns inherent in recombinant angiogenic/growth factors. We believe a combination therapy of the DeBakey VAD-synthetic peptide could provide sufficient revascularization for bridge-to-recovery or as an alternative-to-transplant in certain patients. PMID- 11403674 TI - Development status of Terumo implantable left ventricular assist system. AB - We have been developing an implantable left ventricular assist system (T-ILVAS) featuring a magnetically suspended centrifugal pump (MSCP) since 1995. In vitro and in vivo studies using a prototype MSCP composed of a polycarbonate housing and impeller (196 ml) have demonstrated long-term durability and excellent blood compatibility for up to 864 days, and excellent stability of the magnetic bearing of the MSCP. These preliminary results strongly suggested that the magnetic bearing of the MSCP is reliable and is a most feasible mechanism for a long-term circulatory assist device. We have recently devised a clinical version pump made of titanium (180 ml) with a new position sensor mechanism and a wearable controller with batteries. Cadaver fit study confirmed that the Type IV pump could be implanted in a small patient with a body surface area as small as 1.3. The in vitro performance tests of the Type IV pump demonstrated excellent hydrodynamic performances with an acceptable hemolysis rate. New position sensors for the titanium housing showed more uniform sensor outputs of a magnetic bearing than in the prototype polycarbonate pump. The Type IV pump then was evaluated in vivo in 6 sheep at the Oxford Heart Centre. Four sheep were electively sacrificed at 3 months and were allowed to survive for more than 6 months for long-term evaluation. In this particular series of experiments, no anticoagulant/antiplatelet regimen was utilized except for a bolus dose of heparin during surgery. There was a left ventricular mural thrombi around the inflow cannula in 1 sheep. Otherwise, there was no mechanical failure nor sign of thromboembolism throughout the study. PMID- 11403675 TI - From a lab type to a product: a retrospective view on Impella's assist technology. AB - A lab type is best described by its value as a result of its handcrafted uniqueness in small numbers. Logically, there is not one lab type like another, and the fact that it has been realized does not mean that this special effort can be easily reproduced. Furthermore, most lab types have undergone stand alone test runs revealing fingerprints rather than universal results at a 20% effort to 80% effect ratio. A product development, on the other hand, is best described by an 80% effort to 20% effect ratio in terms of measurable results. Products are producible and cost effective goods which are well documented and have undergone numerous test runs and test procedures assuring safety and quality, a basic requirement for market approval and cost effective marking. Based on the intravascular pump technology, comprising a sensorized axial flow pump with an integrated micromotor, the iterative dependence of the product development on lab types is demonstrated showing in particular the importance of having highly developed lab types before initiating the product development. By example, we demonstrated that high product quality has a greater impact on the reduction of blood damage than numerous redesigns. Reengineering issues are addressed, which are part of the product development process. Furthermore, the previously mentioned technology serves as a platform leading directly from the perioperative biventricular system to a 7 day pump as well as a miniaturized 12 Fr version. PMID- 11403676 TI - HeartMate left ventricular assist devices: a multigeneration of implanted blood pumps. AB - The HeartMate family of implanted left ventricular assist devices (LVADs) developed by Thermo Cardiosystems, Inc. (TCI) span a time frame that goes back to the beginning of clinical use of mechanical circulatory support and will stretch well into the foreseeable future. Associated blood pump technology employed in the HeartMates range from an original pusher plate concept to the most advanced rotary pump devices. Starting initially with a pneumatic actuated pusher plate pump, clinical use of the HeartMate I began in 1986. In 1990, electric motor actuated versions of the HeartMate I began to be used clinically. Presently, the HeartMate I has been implanted in some 2,300 patients worldwide, and this LVAD is a standard by which all others are currently measured. Following the HeartMate I is TCI's next-generation, the HeartMate II, a rotary-pump-based LVAD that uses an axial flow blood pump having blood immersed mechanical bearings. Clinical trials of the HeartMate II were initiated in 2000. The HeartMate III, representing TCI's next-generation LVAD, is structured around a centrifugal blood pump that uses a magnetically levitated rotating assembly. Compared to the HeartMate II, the HeartMate III has the potential for higher overall efficiency. The pump's operating life is not dependent on bearing wear. Given the significantly advanced LVAD technology represented by HeartMates II and III, coupled with the experience of HeartMate I, TCI is well-poised to keep its LVAD products as industry standards in the future. PMID- 11403677 TI - Glutamate transporter expression in astrocytes of the rat dentate gyrus following lesion of the entorhinal cortex. AB - The glutamate transporters GLT-1 and GLAST localized in astrocytes are essential in limiting transmitter signalling and restricting harmful receptor overstimulation. To show changes in the expression of both transporters following lesion of the entorhinal cortex (and degeneration of the glutamatergic tractus perforans), quantitative microscopic in situ hybridization (ISH) using alkaline phosphatase-labelled oligonucleotide probes was applied to the outer molecular layer of the hippocampal dentate gyrus of rats (termination field of the tractus perforans). Four groups of rats were studied: sham-operated controls, and animals 3, 14 and 60 days following unilateral electrolytic lesion of the entorhinal cortex. The postlesional shrinkage of the terminal field of the perforant path, ipsilateral to the lesion side, was determined and considered in the evaluation of quantitative ISH data. Statistical analysis revealed that ipsilateral to the lesion side there was a significant decrease of the GLT-1 mRNA at every postlesional time-point and of the GLAST mRNA at 14 and 60 days postlesion. The maximal decrease was approximately 45% for GLT-1 and approximately 35% for GLAST. In the terminal field of the perforant path contralateral to the lesion side, no significant changes of ISH labelling were measured. The results were complemented by immunocytochemical data achieved using antibodies against synthetic GLT-1 and GLAST peptides. In accordance with ISH results, there was an obvious decrease of GLT-1 and GLAST immunostaining in the terminal field of the perforant path ipsilateral to the lesion side. From these data we conclude that, following a lesioning of the entorhinal cortex, the loss of glutamatergic synapses in the terminal field of the perforant path resulted in a strong downregulation of glutamate transporters in astrocytes. The decrease of synaptically released glutamate or of other neuronal factors could be involved in this downregulation. PMID- 11403678 TI - Neurotoxicity of channel mutations in heterologously expressed alpha7-nicotinic acetylcholine receptors. AB - Nicotinic acetylcholine receptors (nAChR) composed of chick alpha7 subunits mutated to threonine at amino acid valine-251 in the putative channel-lining M2 domain were expressed heterologously in several neuron-like and non-neuronal mammalian cell lines. Expression of mutant alpha7-nAChR is toxic to neuron-like cells of the human neuroblastoma cell lines SH-SY5Y and IMR-32, but not to several other cell types. Growth in the presence of the alpha7-nAChR antagonist methyllycaconitine (MLA) protects against neurotoxicity, as does gradual downregulation of functional, mutant alpha7-nAChR in surviving transfected SH SY5Y cells. Relative to wild-type alpha7-nAChR, functional alpha7-nAChR mutants show a higher affinity for agonists, slower rates of desensitization, and sensitivity to dihydro-beta-erythroidine (DHbetaE) as an agonist, but they retain sensitivity to MLA as a competitive antagonist. These findings demonstrate that expression of hyperfunctional, mutant forms of Ca2+-permeable alpha7-nAChR is toxic to neuron-like cells. PMID- 11403679 TI - Protective effect of harmaline and harmalol against dopamine- and 6 hydroxydopamine-induced oxidative damage of brain mitochondria and synaptosomes, and viability loss of PC12 cells. AB - The present study elucidated the protective effect of beta-carbolines (harmaline, harmalol and harmine) against oxidative damage of brain mitochondria, synaptosomes and PC12 cells induced by either dopamine or 6-hydroxydopamine. Harmaline, harmalol and antioxidant enzymes (superoxide dismutase/SOD and catalase) decreased the alteration of mitochondrial swelling and membrane potential induced by 200 microM dopamine or 100 microM 6-hydroxydopamine. Deprenyl attenuated the dopamine-induced mitochondrial dysfunction but did not reduce the effect of 6-hydroxydopamine. While beta-carbolines inhibited the electron flow in mitochondria, they did not enhance the depressant effect of catecholamines. beta-Carbolines and antioxidant enzymes reversed the depression of synaptosomal Ca2+ uptake induced by 10 microM catecholamines. The compounds inhibited the catecholamine-induced thioredoxin reductase inhibition, thiol oxidation and carbonyl formation in mitochondria and synaptosomes. beta Carbolines decreased the reactive species-induced deoxyribose degradation. Harmaline and harmalol reduced the catecholamine-induced loss of the transmembrane potential and of cell viability in PC12 cells. beta-Carbolines alone did not show a significant cytotoxic effect on PC12 cells. The results suggest that beta-carbolines may attenuate the dopamine- or 6-hydroxydopamine induced alteration of brain mitochondrial and synaptosomal functions, and viability loss in PC12 cells, by a scavenging action on reactive oxygen species and inhibition of thiol oxidation. PMID- 11403680 TI - Patterned cerebellar Purkinje cell death in a transgenic mouse model of Niemann Pick type A/B disease. AB - Niemann Pick disease is a family of autosomal recessive disorders characterized by cholesterol accumulation. The most common type is Niemann Pick type A/B (NPA/B), resulting from deficient acid sphingomyelinase activity, which leads to sphingomyelin and cholesterol accumulation. The neuropathology of NPA/B includes widespread neuronal degeneration. An acid sphingomyelinase knockout mouse model of NPA/B (ASMKO) has been developed by the targeted deletion of the acid sphingomyelinase gene. When cerebellar morphology was examined in the ASMKO mouse at postnatal day 60 (P60), a dramatic pattern of longitudinal stripes was revealed in which roughly half the Purkinje cells had died, leaving a highly stereotyped, bilaterally symmetrical array of stripes. Antizebrin II immunocytochemistry revealed that the absent Purkinje cells corresponded exactly to the zebrin II-negative subset, leaving the zebrin II-positive subset apparently intact. By P120, some of the zebrin II-positive Purkinje cells had also been eliminated from the posterior vermis and hemispheres. By P180, all Purkinje cells had been lost from the anterior lobe. Finally at P240, almost all Purkinje cells had disappeared to leave a stereotyped distribution in lobules VI, IX-X and the flocculus and paraflocculus. The temporal pattern of Purkinje cell death demonstrates differential susceptibility of morphologically identical cells that appear to be linked to their molecular phenotypes. PMID- 11403681 TI - Identification of a cis-acting dendritic targeting element in the mRNA encoding the alpha subunit of Ca2+/calmodulin-dependent protein kinase II. AB - In mammalian neurons a selected group of mRNAs, including the transcript encoding the alpha subunit of Ca2+/calmodulin-dependent protein kinase II, is found in dendrites. The molecular mechanisms underlying extrasomatic RNA trafficking are not well described. It is thought that dendritic transcripts contain cis-acting elements that direct their selective subcellular sorting. Here we report the identification of an extrasomatic targeting element in the 3' untranslated region of the mRNA encoding the alpha subunit of Ca2+/calmodulin-dependent protein kinase II. In primary hippocampal neurons, this 1200-nucleotide-spanning, cis acting element is sufficient to mediate dendritic localization of chimeric reporter transcripts. The trafficking signal does not share any striking sequence similarity with a previously characterized dendritic targeting element in transcripts encoding the microtubule-associated protein 2. In dendrites of transfected primary neurons, recombinant RNAs form granules with an average diameter of 0.45 microm that may represent preferential RNA docking sites or multimolecular transport units. These findings imply that extrasomatic sorting of individual dendritic mRNAs involves at least partially distinct molecular mechanisms, as well as large trafficking complexes. PMID- 11403682 TI - Efferent axons in the avian auditory nerve. AB - The sensory hair cells of the inner ear receive both afferent and efferent innervation. The efferent supply to the auditory organ has evolved in birds and mammals into a separate complex system, with several types of neurons of largely unknown function. In this study, the efferent axons in four different species of birds (chicken, starling, barn owl and emu) were examined anatomically. Total numbers of efferents supplying the cochlear duct (auditory basilar papilla and the vestibular lagenar macula) were determined; separate estimates of the efferents to the lagenar macula only were also derived and subtracted. The numbers for auditory efferents thus varied between 120 (chicken) and 1068 (barn owl). Considering the much larger numbers of hair cells in the basilar papilla, each efferent is predicted to branch extensively. However, pronounced species specific differences as well as regional differences along the tonotopic gradient of the basilar papilla were documented. Myelinated and unmyelinated axons were found, with mean diameters of about 1 microm and about 0.5 microm, respectively. This suggests two basic populations of efferents, however, they did not appear to be distinguished sharply. Evidence is presented that some efferents lose their myelination at the transition from central oligodendrocyte to peripheral Schwann cell myelin. Finally, a comparison of the four bird species evaluated suggests that the efferent population with smaller, unmyelinated axons is the phylogenetically more primitive one. A new population probably arose in parallel with the evolution and differentiation of the specialized hair-cell type it innervates, the short hair cell. PMID- 11403683 TI - The Cav2.1/alpha1A (P/Q-type) voltage-dependent calcium channel mediates inhibitory neurotransmission onto mouse cerebellar Purkinje cells. AB - The effects of voltage-dependent calcium channel (VDCC) antagonists on spontaneous inhibitory postsynaptic currents (sIPSCs) in mouse Purkinje cells were examined using in vitro cerebellar slices. The inorganic ion Cd2+ reduced sIPSC amplitude and frequency. No additional block was seen with the Na+ channel antagonist tetrodotoxin (TTX) suggesting that all action potential-evoked inhibitory GABA release was mediated by high-voltage-activated VDCCs. No evidence was found for involvement of Cav1/alpha1C and alpha1D (L-type), Cav2.2/alpha1B (N type) or Cav2.3/alpha1E (R-type) high-voltage-activated VDCCs or low-voltage activated Cav3/alpha1G, alpha1H and alpha1I (T-type) VDCCs in mediating presynaptic GABA release. Blockade of sIPSCs by 200 nM omega-agatoxin IVA implicated the Cav2.1/alpha1A (P/Q-type) subtype of high-voltage-activated VDCCs in mediating inhibitory transmission. Inhibition by omega-agatoxin IVA was similar to that seen with Cd2+ and TTX. Selective antibodies directed against the Cav2.1 subunit revealed staining in regions closely opposed to Purkinje cell somata. Cav2.1 staining was colocalized with staining for antibodies against glutamic acid decarboxylase and corresponded well with the pericellular network formed by GABAergic basket cell interneurons. Antibody labelling of Cav2.3 revealed a region-specific expression. In the cerebellar cortex anterior lobe, Cav2.3 staining was predominantly somatodendritic; whilst in the posterior lobe, perisomatic staining was seen primarily. However, electrophysiological data was not consistent with a role for the Cav2.3 subunit in mediating presynaptic GABA release. No consistent staining was seen for other Cav (alpha1) subunits. Electrophysiological and immunostaining data support a predominant role for Cav2.1 subunits in mediating action potential-evoked inhibitory GABA release onto mouse Purkinje cells. PMID- 11403684 TI - Activation of glycogen synthase kinase 3 beta (GSK-3beta) by platelet activating factor mediates migration and cell death in cerebellar granule neurons. AB - Children with vertically acquired HIV-1 can present with a rapidly progressive encephalopathy and neuronal apoptosis in the first 12-18 months of life. Furthermore, abnormal prenatal platelet activating factor (PAF) signalling may result in lissencephaly, a disorder of neuronal migration. PAF, produced from human immunodeficiency virus type 1 (HIV-1) -infected brain-resident macrophages, induces neuronal apoptosis in cultured cerebellar granule neurons (CGNs) in part by activating glycogen synthase kinase 3 beta (GSK-3beta). However, PAF can also inhibit migration of CGNs that are dispersed and allowed to reaggregate. Therefore, we investigated the biological effects following activation of GSK 3beta by PAF, and whether these effects were dependent on the culture conditions of the CGNs. We show here that activation of neuronal GSK-3beta by PAF is receptor-specific, with similar kinetics of activation in both monolayer cultures of CGNs that have ceased to migrate and reaggregate cultures of CGNs that are actively migrating. However, PAF receptor activation in reaggregated CGNs inhibits neuronal migration and induces approximately half the level of neuronal apoptosis compared with PAF-treated CGN cultures that have ceased to migrate. PAF mediated inhibition of neuronal migration in reaggregated CGNs or induction of apoptosis in CGNs that have ceased to migrate can be reversed by either PAF receptor antagonists, or the GSK-3beta inhibitors lithium or valproic acid, in a dose-dependent manner. Abnormal PAF signalling that results in GSK-3beta overactivation may represent a common mechanism for pathological defects in neuronal migration in the prenatal period and neuronal apoptosis in the postnatal period. PMID- 11403685 TI - A single administration of interleukin-1 or amphetamine induces long-lasting increases in evoked noradrenaline release in the hypothalamus and sensitization of ACTH and corticosterone responses in rats. AB - Single administration of the cytokine interleukin-1beta (IL-1) or the psychostimulant amphetamine causes long-term sensitization of the hypothalamus pituitary adrenal (HPA) axis, i.e. enhanced adrenocorticotropine hormone (ACTH) and corticosterone responses weeks later. HPA responses to these stimuli involve activation of hypothalamic corticotropin-releasing hormone (CRH) neurons by noradrenergic projections to the paraventricular nucleus (PVN). In search of the underlying mechanisms, we studied the temporal pattern of HPA sensitization in relation to (1) the reactivity of noradrenergic projections to the PVN and (2) altered secretagogue production in hypothalamic CRH neurons. Single exposure to IL-1 or amphetamine induced cross-sensitization of ACTH and corticosterone responses 11 and 22 days later, but not after 42 days. Amphetamine-induced HPA sensitization was not accompanied by increased costorage of arginine vasopressin (AVP) in CRH terminals, as found previously after IL-1 pretreatment. The reactivity of noradrenergic terminals was assessed by measuring the electrically evoked release of [3H]-noradrenaline from superfused PVN slices. Single administration of amphetamine and IL-1 induced a long-lasting (up to 22 days) increase (up to 165%) of evoked noradrenaline release. This indicates that single exposure to psychostimulants or to cytokines can induce a long-lasting increase in stimulus-secretion coupling in brainstem noradrenergic neurons that innervate the PVN. This common, long-lasting functional change may underlie, at least in part, IL-1- and amphetamine-induced HPA cross-sensitization. In addition, increased AVP signalling by hypothalamic CRH neurons appears to play a role in IL 1-induced, but not in amphetamine-induced, HPA sensitization. PMID- 11403686 TI - Dopaminergic retinal cell differentiation in culture: modulation by forskolin and dopamine. AB - We examined the effects of dopamine and cAMP on the differentiation of dopaminergic retinal cells in the chick retina, using an in vitro system and tyrosine hydroxylase immunocytochemistry. Tyrosine hydroxylase-positive cells were detected in cultures prepared from embryonic day 10 retinas. These increased in number as a function of time in vitro and by treatment for 4 days with forskolin. Besides causing a 3.4-fold increase in the tyrosine hydroxylase positive population, forskolin also caused these cells to developed morphogenetic features of more mature cells. As opposed to forskolin, cultures treated with dopamine exhibited a 55% reduction of the tyrosine hydroxylase-positive cell population, as compared to untreated cultures. Quinpirole was able to mimic the dopamine effect. This dopamine effect could only be blocked by clozapine, whereas raclopride and eticlopride were ineffective. Our results suggest the existence of a narrow window during development when undifferentiated dopaminergic cells are capable of being influenced by specific signals, possibly via cAMP production. The data also indicate that dopamine may act as a regulatory factor limiting the tyrosine hydroxylase-positive population in the retina. PMID- 11403687 TI - Differentiation-dependent sensitivity to cell death induced in the developing retina by inhibitors of the ubiquitin-proteasome proteolytic pathway. AB - The effects of inhibitors of proteasome function were studied in the retina of developing rats. Explants from the retina of neonatal rats at postnatal day (P) 3 or P6 were incubated with various combinations of the proteasome inhibitor carbobenzoxyl-leucinyl-leucinyl-leucinal (MG132), the protein synthesis inhibitor anisomycin, or the adenylyl cyclase activator forskolin. MG132 induced cell death in a subset of cells within the neuroblastic (proliferative) layer of the retinal tissue. The cells sensitive to degeneration induced by either MG132 or anisomycin, were birthdated by bromodeoxyuridine injections. This showed that the MG132-sensitive population includes both proliferating cells most likely in their last round of cell division, and postmitotic undifferentiated cells, at a slightly earlier stage than the population, sensitive to anisomycin-induced cell death. The results show that sensitivity to cell death induced by proteasome inhibitors defines a window of development in the transition from the cell cycle to the differentiated state in retinal cells. PMID- 11403688 TI - Deficient long-term synaptic depression in the rostral cerebellum correlated with impaired motor learning in phospholipase C beta4 mutant mice. AB - Long-term depression (LTD) at parallel fibre-Purkinje cell synapse of the cerebellum is thought to be a cellular substrate for motor learning. LTD requires activation of metabotropic glutamate receptor subtype 1 (mGluR1) and its downstream signalling pathways, which invariably involves phospholipase Cbetas (PLCbetas). PLCbetas consist of four isoforms (PLCbeta1-4) among which PLCbeta4 is the major isoform in most Purkinje cells in the rostral cerebellum (lobule 1 to the rostral half of lobule 6). We studied mutant mice deficient in PLCbeta4, and found that LTD was deficient in the rostral but not in the caudal cerebellum of the mutant. Basic properties of parallel fibre-Purkinje cell synapses and voltage-gated Ca2+ channel currents appeared normal. The mGluR1-mediated Ca2+ release induced by repetitive parallel fibre stimulation was absent in the rostral cerebellum of the mutant, suggesting that their LTD lesion was due to the defect in the mGluR1-mediated signalling in Purkinje cells. Importantly, the eyeblink conditioning, a simple form of discrete motor learning, was severely impaired in PLCbeta4 mutant mice. Wild-type mice developed the conditioned eyeblink response, when pairs of the conditioned stimulus (tone) and the unconditioned stimulus (periorbital shock) were repeatedly applied. In contrast, PLCbeta4 mutant mice could not learn the association between the conditioned and unconditioned stimuli, although their behavioural responses to the tone or to the periorbital shock appeared normal. These results strongly suggest that PLCbeta4 is essential for LTD in the rostral cerebellum, which may be required for the acuisition of the conditioned eyeblink response. PMID- 11403689 TI - Opposing effects of behavioural activity and light on neurons of the suprachiasmatic nucleus. AB - The mammalian circadian pacemaker is located in the suprachiasmatic nuclei. It can be shifted in phase by photic cues and by the behavioural activity of the animal. When presented together, light and behavioural activity attenuate each others' phase-shifting effect. Still unclear is how behavioural activity affects the suprachiasmatic nuclei and how it interacts with photic information. Previously, we reported the occurrence of behaviourally induced suppressions of neuronal activity. The present study investigates the characteristics of these suppressions as a function of circadian time and, additionally, in the presence of photic cues. We performed long-term multiunit activity recordings of neurons in freely moving rats and found that these suppressions of neuronal firing in the suprachiasmatic nucleus occurred at every phase of the circadian cycle. The magnitude of the suppressions showed a circadian variation, with larger suppressions during subjective day. When a light pulse was applied during a suppression, light and activity appeared to oppose each others' effects within the recorded population of neurons. The resulting discharge level appeared to be the sum of both responses. The opposing effects of light and activity were also found in single unit recordings, indicating that photic and behavioural stimuli interact at the level of a single neuron. PMID- 11403690 TI - Lesions of the pedunculopontine tegmental nucleus reduce paradoxical sleep (PS) propensity: evidence from a short-term PS deprivation study in rats. AB - Cholinergic neurons in the mesopontine tegmentum are thought to play a critical role in the generation of paradoxical sleep (PS). However, no study has yet examined whether lesions of these neurons cause deficits of PS in the rat. We describe here the effects of lesions of the pedunculopontine tegmental nucleus (PPT) on spontaneous PS and on PS propensity, expressed during and after a short period of PS deprivation. Lesions were induced by bilateral injections of ibotenate. PS deprivation was performed manually by gently waking rats each time they showed polygraphic signs of PS. Two weeks after lesions, an 8-h baseline recording was performed; the following day, rats were PS deprived for 6 h and polygraphic recordings were then continued for 2 h, to examine recovery sleep. The same protocol was repeated 1 week later. Compared with controls and with rats with limited PPT lesions, rats bearing > 60% NADPH-diaphorase-positive cell loss within the PPT showed unaffected PS under baseline conditions. However, they made fewer attempts to enter PS during deprivation and they exhibited an attenuated rebound increase in PS time after deprivation. The number of PS attempts and the magnitude of PS rebound were negatively correlated with the percent loss of diaphorase-positive neurons within the PPT. Thus, PS propensity that accumulated as a result of PS deprivation was reduced after extensive PPT lesions. In summary, although spontaneous PS was found to be unaltered, the PS deprivation procedure used in this study demonstrated the dysfunctioning of PS caused by PPT lesions. PMID- 11403691 TI - Amphetamine and cocaine induce different patterns of c-fos mRNA expression in the striatum and subthalamic nucleus depending on environmental context. AB - In the dorsal striatum, there are two major populations of medium spiny projection neurons. One population is positive for dynorphin mRNA (DYN+), and these cells project preferentially to the substantia nigra, forming the so-called 'direct pathway'. A second population is positive for enkephalin mRNA (ENK+), and these cells influence the substantia nigra indirectly, via the globus pallidus and subthalamic nucleus. Psychostimulant drugs, such as amphetamine and cocaine, are reported to induce immediate early genes (IEGs) in only one subpopulation of dorsal striatal projection neurons, DYN+ cells. However, this apparent selectivity appears to be a function of environmental context. We found that when given in the animal's home cage, amphetamine and cocaine increased expression of the IEG, c-fos, almost exclusively in DYN+ cells. However, when given in a novel environment, amphetamine and cocaine increased c-fos mRNA in both DYN+ and ENK+ cells. Furthermore, amphetamine and cocaine increased c-fos mRNA expression in the subthalamic nucleus when administered in the novel environment, but not when given at home. We conclude that the neural circuitry engaged by psychostimulant drugs, and their ability to induce specific patterns of gene expression, are determined by the environmental context in which they are experienced. This may be related to the ability of environmental novelty to facilitate psychostimulant drug-induced neuroplasticity. PMID- 11403692 TI - Involvement of the central nucleus of the amygdala and nucleus accumbens core in mediating Pavlovian influences on instrumental behaviour. AB - Pavlovian conditioned cues exert a powerful influence on instrumental actions directed towards a common reward, this is known as Pavlovian-to-instrumental transfer (PIT). The nucleus accumbens (NAcc) has been hypothesized to function as an interface between limbic cortical structures required for associative conditioning, like the amygdala, and response mechanisms through which instrumental behaviour can be selected and performed. Here we have used selective excitotoxic lesions to investigate the involvement of subnuclei of the amygdala as well as the core and shell regions of the nucleus accumbens on PIT in rats. Within the amygdala, selective lesions of the central nucleus (CeN), but not of the basolateral nucleus (BLA), abolished the PIT effect. In addition, selective lesions of the NAcc core, but not the NAcc shell, also abolished PIT. None of the lesions impaired the acquisition of Pavlovian food cup approaches or instrumental responding itself. These data demonstrate that the CeN and NAcc core are central components of the neural system mediating the impact of Pavlovian cues on instrumental responding. We suggest that this effect may depend upon the regulation of the dopaminergic innervation of the NAcc core by projections from the CeN to the ventral tegmental area. PMID- 11403693 TI - Daily variation and appetitive conditioning-induced plasticity of auditory cortex receptive fields. AB - Long-term modification of cortical receptive field maps follows learning of sensory discriminations and conditioned associations. In the process of determining whether appetitive - as opposed to aversive - conditioning is effective in causing such plastic changes, it was discovered that multineuron receptive fields, when measured in rats under ketamine-sedation, vary substantially over the course of a week, even in the absence of classical conditioning and electrode movement. Specifically, a simple correlation analysis showed that iso-intensity frequency response curves of multiunit clusters and local field potentials recorded from auditory cortex are nonstationary over 7 days. Nevertheless, significant plastic changes in receptive fields, due to conditioned pairing of a pure tone and electrical stimulation of brain reward centres, are detectable above and beyond these spontaneous daily variations. This finding is based on a novel statistical plasticity criterion which compares receptive fields recorded for three days before and three days after conditioning. Based on a more traditional criterion (i.e. one day before and after conditioning), the prevalence of learning-induced changes caused by appetitive conditioning appears to be comparable to that described in previous studies involving aversive conditioning. PMID- 11403694 TI - Purely temporal figure-ground segregation. AB - Visual figure-ground segregation is achieved by exploiting differences in features such as luminance, colour, motion or presentation time between a figure and its surround. Here we determine the shortest delay times required for figure ground segregation based on purely temporal features. Previous studies usually employed stimulus onset asynchronies between figure- and ground-containing possible artefacts based on apparent motion cues or on luminance differences. Our stimuli systematically avoid these artefacts by constantly showing 20 x 20 'colons' that flip by 90 degrees around their midpoints at constant time intervals. Colons constituting the background flip in-phase whereas those constituting the target flip with a phase delay. We tested the impact of frequency modulation and phase reduction on target detection. Younger subjects performed well above chance even at temporal delays as short as 13 ms, whilst older subjects required up to three times longer delays in some conditions. Figure-ground segregation can rely on purely temporal delays down to around 10 ms even in the absence of luminance and motion artefacts, indicating a temporal precision of cortical information processing almost an order of magnitude lower than the one required for some models of feature binding in the visual cortex [e.g. Singer, W. (1999), Curr. Opin. Neurobiol., 9, 189-194]. Hence, in our experiment, observers are unable to use temporal stimulus features with the precision required for these models. PMID- 11403695 TI - Post-weaning social isolation of rats leads to an abnormal gait. AB - The purpose of this study was to further characterize the phenotype of rats that have experienced prolonged postweaning social isolation, a paradigm that produces changes relevant to neuropsychiatric disorders. At weaning, male Wistar rats from three litters were housed socially (n = 12) or in isolation (n = 13) for 10 weeks. Isolated rats could see, hear and smell other rats. A sophisticated analysis of gait revealed that the stride duration was increased in isolates (12%, P = 0.0024) as a result of increased stance duration (18%, P = 0.0005), but there was no difference in vertical reaction force or velocity. Adrenal glands were heavier in isolates (19%, P = 0.0047). There was no difference in cross sectional area of the brain or lateral ventricles anywhere along the anterior posterior axis. All experiments and analysis were performed blind to housing condition. This is the first study to demonstrate that socially isolated rats have an abnormal gait. Further analysis, including pharmacological manipulation, is needed in order to understand the nature of the abnormality. PMID- 11403697 TI - Acute exacerbation of idiopathic pulmonary fibrosis and fibroproliferative acute respiratory distress syndrome: similar disease, similar treatment? PMID- 11403698 TI - Evaluation of inspiratory rise time and inspiration termination criteria in new generation mechanical ventilators: a lung model study. AB - INTRODUCTION: Inspiratory rise time adjustment during pressure ventilation and inspiration termination criteria adjustment during pressure support ventilation are available on some of the newest mechanical ventilators. Both are designed to improve patient-ventilator synchrony. However, the function of these adjuncts during pressure ventilation on these ventilators has not been evaluated. METHODS: Three inspiratory rise times (minimum, medium, and maximum) were evaluated in 5 new-generation mechanical ventilators (Hamilton Galileo, Siemens 300A, Puritan Bennett 840, BEAR 1000, and Drager Evita 4) during pressure support and pressure assist/control. Three inspiration termination criteria settings (minimum, medium, and maximum) were also evaluated in 2 mechanical ventilators (Hamilton Galileo and Puritan Bennett 840) during pressure support. All evaluations were performed with a spontaneous breathing lung model (compliance 50 mL/cm H2O, resistance 8.2 cm H2O/L/s, respiratory rate 12 breaths/min, inspiratory time 1.0 s, and lung model peak inspiratory flow 60 L/min). Throughout the evaluation, inspiratory pressure was set at 15 cm H2O and positive end-expiratory pressure at 5 cm H2O, resulting in a peak airway pressure of 20 cm H2O. RESULTS: Significant (p < 0.05) and important (> 10%) differences were found among the ventilators at similar rise times (minimum, medium, and maximum) and for each ventilator as rise time was varied. Also, significant (p < 0.05) and important (> 10%) differences were observed between ventilators and within each ventilator when inspiration termination criteria were varied. There were significant (p < 0.05) differences between pressure support and pressure assist/control, but most were < 10%, except those associated with expiration. CONCLUSIONS: Major differences exist for each ventilator as rise time or inspiration termination criteria are varied and among ventilators at similar settings. Inspiration termination criteria adjustment markedly affects transition to exhalation in the Puritan Bennett 840. PMID- 11403699 TI - Mean alveolar pressure during constant-flow and constant-pressure inflation of diseased lungs. AB - INTRODUCTION: An important goal in managing mechanical ventilation is optimizing key variables such as mean alveolar pressure (PA). PURPOSE: Determine the effects of lung mechanics on PA during constant-flow inflation (CFI) and constant pressure inflation (CPI) in models of nonhomogenous lung disease. METHODS: We postulated a mathematical lung model consisting of 2 parallel lung units with the airways and the chest wall. Analysis was performed for step inputs of inspiratory flow and pressure while maintaining the same tidal volume, respiratory frequency, and positive end-expiratory pressure. The analysis was performed for purely restrictive, purely obstructive, and mixed lung disease. RESULTS: When simulating purely restrictive lung disease, PA was always higher in healthy alveoli than in diseased alveoli, with constant inspiratory flow or constant inspiratory pressure. However, PA in each alveoli was always greater during CPI than during CFI. For purely obstructive lung diseases, PA was always lower in diseased alveoli than in healthy alveoli, with constant inspiratory flow or constant inspiratory pressure. However, PA in each alveoli was always greater during CPI than during CFI. For mixed lung diseases with equal time constants, PA was always higher in diseased alveoli than in healthy alveoli, with constant inspiratory flow or constant inspiratory pressure. However, PA in each alveoli was always the same during CPI as during CFI. CONCLUSIONS: For the same tidal volume, the mean alveolar pressure in different alveoli depends on the type of disease. The difference in mean alveolar pressure between a normal and a diseased alveolus depends on the difference in time constant between those alveoli, regardless of the mode of ventilation. PMID- 11403700 TI - Staffing exercise sessions in pulmonary rehabilitation. AB - INTRODUCTION: There is no published standard for the ratio of staff to patients during exercise sessions in outpatient pulmonary rehabilitation. This lack of a standard raises concern about patient safety and the ability to monitor patients adequately. OBJECTIVE: Determine a staff-to-patient ratio standard by examining current practice. METHODS: The directors of the 46 outpatient pulmonary rehabilitation programs in Ohio were surveyed. Directors of 40 (87%) programs responded, and the responses from the 36 programs that conducted pulmonary exercise sessions separately from cardiac sessions were analyzed. RESULTS: The maximum number of staff in an exercise session was 2, with a median of 1. The maximum number of patients in a session was 20, with median of 4. Directors were also asked to rank the importance of 8 factors in determining the reported ratio. From highest to lowest importance, the factors were patient safety, severity of pulmonary disease, availability of space or equipment, entry test data, pulmonary diagnosis, presence of cardiac problems, convenience, and other. Sixty percent of programs restricted the number of patients per session based on staff-to-patient ratio. CONCLUSIONS: Statewide practice for staff-to-patient ratio in Ohio was described. The most common staff-to-patient ratio during exercise sessions was 1:4 or less (78%). Exercise staff work with small groups in order to assure patient safety, to deal with patients' health problems, and to provide adequate facilities. A majority of programs restrict the number of patients per session so as to enforce the reported ratio. PMID- 11403701 TI - Effect of prolonged low-dose methylprednisolone therapy in acute exacerbation of idiopathic pulmonary fibrosis. AB - We report on a 74-year-old man with an acute exacerbation of idiopathic pulmonary fibrosis (IPF) who was successfully treated with prolonged low-dose methylprednisolone, initiated at a loading dose of 2 mg/kg, followed by 2 mg/kg per day for 14 days. The dose was then tapered. The exacerbation observed on the chest radiograph and high-resolution computed tomography was found to have abated after the treatment. This successful case suggests the feasibility of this methylprednisolone treatment protocol for patients with IPF accompanied by accelerated deterioration. Furthermore, this case suggests possible similarities between acute exacerbation of IPF and late acute respiratory distress syndrome, as the same treatment protocol was previously proved to be beneficial for patients with late acute respiratory distress syndrome. PMID- 11403702 TI - Co-existing granular cell tumor and adenocarcinoma of the lung: a case report and review of the literature. AB - Granular cell tumor (GCT) of the lung is a rare tumor, constituting less than 10% of all GCTs. It is a multicentric tumor and infiltrates into adjoining tissue, but malignant GCT of the lung has not been reported. Diagnosis is usually obtained with bronchoscopic biopsy. Treatment options include bronchoscopic extirpation, laser therapy, and sleeve resection. We present a case of GCT co existing with adenocarcinoma of the lung and review the literature. PMID- 11403703 TI - The future of respiratory care. AB - The term respiratory care has more than one meaning, referring both to a subject area within clinical medicine and to a distinct health care profession. In the light of several fundamental transformations of health care during the 20th century, this article reviews the history of respiratory care in both of these contexts and offers 10 predictions for the future: (1) Less focus on raising P(aO2) as a primary goal in managing patients with acute hypoxemic respiratory failure. (2) More attention to the adequacy of tissue oxygenation in such patients, irrespective of P(aO2), and the emergence of "permissive hypoxemia," analogous to permissive hypercapnia, in managing them. (3) Smarter monitors that display information less but process it more, while interacting directly with ventilators and other devices to modify therapeutic interventions. (4) Increased use of and expertise with noninvasive ventilation, with a corresponding decrease in intubations and complications, in treating patients with acute exacerbations of COPD. (5) Increased use of triage in the intensive care unit, including earlier determination of the appropriateness of maximal supportive intervention. (6) Greater use of protocols in patient assessment and management, in all clinical settings. (7) Increased awareness of, expertise in, and resources for palliative care, with a more active and acknowledged role for respiratory therapists. (8) Accelerating progress in smoking cessation and prevention, and also in early detection and intervention in COPD, led by the respiratory care profession. (9) An increasing presence and impact of respiratory therapists as coordinators and care givers in home care. (10) A continued and enlarging role for the journal Respiratory Care in disseminating research findings, clinical practice guidelines, protocols, and practical educational materials in all areas of the field. PMID- 11403704 TI - Radiosensitivity of DNA in a specific protein-DNA complex: the lac repressor-lac operator complex. AB - PURPOSE: To calculate the probability of radiation-induced frank strand breakage (FSB) at each nucleotide in the Escherichia coli lac repressor-lac operator system using a simulation procedure. To compare calculated and experimental results. To asses the contribution of DNA conformational changes and of the masking by the protein to DNA protection by the repressor. MATERIALS AND METHODS: Two structures of the complex were extracted from the PDB databank: crystallography- and NMR-based structures. Calculations were made of the accessibility of the atoms mainly involved in strand breakage (H4' and H5') to O&Hdot; and of the FSB probabilities, along: (1) DNA in the complex; (2) DNA in the complex depleted of the repressor; and (3) a linear DNA having the same sequence. An 80bp fragment bearing the operator was irradiated alone or in presence of the repressor. The relative probabilities of FSB at each nucleotide were determined using sequencing gel electrophoresis. RESULTS: Calculations predict modulation of the accessibility of H4' and H5' atoms and of the probabilities of FSB along the DNA fragments of complexes. This is due to the protein-induced conformational change and to masking by bound protein. The best agreement with the experimental FSB was observed for calculations that use the crystallography-based structure. CONCLUSIONS: For specific DNA-protein complexes, our calculations can predict the protein radiolytic footprints on DNA. They show the significant contribution of the protein-induced DNA conformational change to DNA protection. PMID- 11403705 TI - Low-dose hypersensitivity after fractionated low-dose irradiation in vitro. AB - PURPOSE: It was demonstrated previously that some radioresistant tumour cell lines respond to decreasing single, low radiation doses by becoming increasingly radiosensitive. This paper reports the response of four radioresistant human glioma cell lines to multiple low-dose radiation exposures given at various intervals. Three of the cell lines (T98G, U87, A7) were proven already to show low-dose hyper-radiosensitivity (HRS) after single low doses; the fourth, U373, does not show HRS after acute doses. MATERIALS AND METHODS: Clonogenic cell survival measurements were made in vitro using the Dynamic Microscopic Image Processing Scanner (DMIPS) or Cell Sorter (CS) following exposure to 240kVp X rays one or more times. RESULTS: A consistent, time-dependent hypersensitive response to a second, or subsequent, dose was observed in the cell lines that demonstrated HRS. This time-dependent change in radiosensitivity did not occur in the radioresistant cell line that did not show HRS (U373). In one cell line that demonstrated strong HRS, T98G, a similar time-dependent hypersensitive response was also seen when the cells were irradiated whilst held in the G1-phase of the cell cycle. In this same cell line, significantly increased cell kill was demonstrated when three very low doses (0.4 Gy) were given per day, 4 h apart, for 5 days, compared with the same total dose given as once-daily 1.2Gy fractions. CONCLUSIONS: These data demonstrate the possibility that a multipledose per day, low-dose per fraction regimen, termed 'ultrafractionation', could produce increased tumour cell kill in radioresistant tumours compared with the same total dose given as conventional-sized 2 Gy fractions. PMID- 11403706 TI - Tumorigenic target cell regions in bone marrow studied by localized dosimetry of 239Pu, 241Am and 233U in the mouse femur. AB - PURPOSE: To study the temporal change in microdistribution of plutonium-239, americium-241 and uranium-233 in the mouse distal femur and to compare and combine calculated radiation doses with those obtained previously for the femoral shaft. Also, to relate doses to relative risks of osteosarcoma and acute myeloid leukaemia. MATERIALS AND METHODS: Computer-based image analysis of neutron induced and alpha-track autoradiographs of sections of mouse femora was used to quantify the microdistribution of (239)Pu, (241)Am and (233)U from 1 to 448 days after intraperitoneal injection. Localized dose-rates and cumulative doses over this period were calculated for different regions of the marrow spaces in trabecular bone. The results were then combined with previous data for doses to the cortical marrow of the femoral shaft. A morphometric analysis of the distal femur was carried out. RESULTS: Initial deposition on endosteal surfaces and dose rates near to the trabecular surfaces at 1 day were two to four times greater than corresponding results for cortical bone. Burial was most rapid for (233)U, about twice the rate in cortical bone. As in cortical bone, subsequent uptake into the marrow was seen for (239)Pu and (241)Am but not (233)U. Cumulative doses to 448 days for different regions of trabecular marrow were greater than corresponding values for cortical marrow for each radionuclide. Combined doses reflected the greater overall volume of cortical marrow. CONCLUSIONS: Cumulative radiation doses to the 10 microm thick band of marrow adjacent to all endosteal surfaces were in the ratio of approximately 7:3:1 for (239)Pu:(241)Am:(233)U. This ratio is not inconsistent with observed incidences of osteosarcoma induction by the three nuclides. Analysis of doses to different depths of marrow, however, showed that although ratios were probably not significantly different to that for a 10 microm depth, better correlations with osteosarcomagenic risk were obtained with 20-40 microm depths. For acute myeloid leukaemia, the closest relationship between relative risk and doses was obtained by considering only the central 5 10% of marrow, which gave a dose ratio of approximately 12:11:1 for (239)Pu:(241)Am:(233)U respectively. PMID- 11403707 TI - Translocation frequencies measured in patients one year after radioactive iodine therapy for thyrotoxicosis. AB - PURPOSE: To investigate the incidence of translocations induced by iodine-131 therapy in thyrotoxicosis patients 1 year after the administration of the radiolabelled compound. MATERIALS AND METHODS: Tricolour FISH with whole chromosome-specific probes for chromosomes 2, 4 and 8 was used for scoring translocations. From the genomic translocation frequencies, derived using the Lucas formula, equivalent whole-body doses were calculated, based on the in vitro (60)Co gamma-ray dose-response curve. RESULTS: A total of 101 translocations were observed in 4864 metaphases, 63% being of the two-way type. In the control group used for obtaining dose-response data, nine translocations were observed in 5278 metaphases, 55% being two-way translocations. No correlation was found between the observed frequency of translocations and administered radioactivity. Using the in vitro dose-response, an estimated average dose for the group of nine patients of 0.79 +/- 0.22Gy was obtained. Compared with frequencies following the assumption that the involvement of a particular chromosome in a two-break exchange-type aberration is proportional to its DNA content, chromosome 4 was more frequently involved and chromosomes 2 and 8 less frequently involved in chromosomal rearrangements. CONCLUSION: This study shows that (131)I therapy for thyrotoxicosis patients induced translocations, especially in chromosome 4, which could be detected 1 year after the administration of the radiolabelled compound. PMID- 11403708 TI - Microsatellite instability in acute myelocytic leukaemia developed from A-bomb survivors. AB - PURPOSE: Genetic alterations, including microsatellite instability (MSI), are ultimate steps toward malignant process. To investigate MSI in A-bomb survivors, leukaemic cells were analysed from 13 acute myelocytic leukaemia patients with a history of radiation exposure and also in 12 de novo patients. MATERIALS AND METHODS: To assess the microsatellite changes, a fluorescent system in 10 loci (BAT40, D3S643, D5S107, IRF1, MYC, D9S171, WT1, TP53, DM, D17S855) was used. RESULTS: MSI analysis revealed a high frequency of multiple microsatellite changes in the exposed patients (84.6%) compared with non-exposed patients (8.3%). There was a significant difference (p < 0.001) between the two groups. CONCLUSIONS: These analyses clearly demonstrate that leukaemic cells from heavily exposed patients contain a number of genetic instabilities that may strongly influence the development of leukaemia among people exposed to the Hiroshima A bomb radiation. PMID- 11403709 TI - Modelling the dose-volume response of the spinal cord, based on the idea of damage to contiguous functional subunits. AB - PURPOSE: To investigate the response of the spinal cord of experimental animals to homogeneous irradiation, the main purpose being to propose a new version of the Critical Volume Normal Tissue Complication Probability (NTCP) model, incorporating spatial correlation between damaged functional subunits (FSU). METHOD: The standard Critical Volume NTCP model and its modified version, the Contiguous Damage model promoted here, are described in mathematical terms. Also, a fiber-like structure of the spinal cord is considered, which is a more complex structure than the standard Critical Volume NTCP model assumes. It is demonstrated that the Contiguous Damage model predicts different responses to two segment irradiation and to single-segment irradiation to the same combined length as observed in experiments on rats, a result that cannot be described by the standard Critical Volume NTCP model. RESULTS AND CONCLUSIONS: Both the Critical Volume model and the Contiguous Damage model, are fitted to two sets of canine spinal cord radiation data corresponding to two different fractionation regimes of irradiation. Whole-organ irradiation as well as partial irradiation to different lengths are considered, allowing the investigation of dose-volume effects. Formal goodness-of-fit investigation shows that both models fit the canine spinal cord data equally well. PMID- 11403710 TI - Level of Flt3-ligand in plasma: a possible new bio-indicator for radiation induced aplasia. AB - PURPOSE: To follow plasma Flt3-ligand (FL) concentrations in irradiated animals in order to evaluate it as an indicator of bone marrow damage for the management of accidental radiation-induced aplasia. MATERIALS AND METHODS: Non-human primates were irradiated at doses ranging from 2 to 8 Gy, using whole- or partial body irradiation. Plasma FL concentrations and blood cell counts were determined daily. RESULTS: FL concentrations increased as early as day 2 after irradiation, whatever the irradiation dose. Increase in plasma FL concentration on day 5 post irradiation was correlated with radiation dose and with the severity of radiation induced aplasia. During the course of aplasia, FL concentrations in plasma were inversely correlated with neutrophil counts. A peak in FL concentration appeared before the neutrophil nadir, and the subsequent decrease in FL concentration was correlated with the recovery of blood-cell populations. CONCLUSIONS: Monitoring plasma FL concentration can be used as an indicator of radiation-induced marrow aplasia, and this may be of use in accidental irradiation situations. PMID- 11403711 TI - Changes of fibrosis-related parameters after high- and low-LET irradiation of fibroblasts. AB - PURPOSE: To investigate the radiation-induced, premature terminal differentiation and collagen production of fibroblasts after heavy ion irradiation. These endpoints are discussed as an underlying cellular mechanism of fibrosis. MATERIALS AND METHODS: Normal human foreskin fibroblasts (AG1522B) were used to determine clonogenic survival, the premature differentiation and synthesis of extracellular matrix (ECM) proteins, e.g. collagen after irradiation with X-rays, 195 and 11.0 MeV u(-1) carbon ions and 9.9 MeV u(-1) nickel ions. Additionally, biopsies from the skin of minipigs were taken. Similar experiments were carried out after irradiation with X-rays and 195 MeV u(-1) carbon ions. RESULTS AND CONCLUSIONS: RBE for clonogenic survival as well as for fibrosis-related parameters for high-energy carbon ions are slightly above unity. Low-energy carbon ions with a higher LET are more efficient than X-rays, whereas the RBE of nickel ions is below unity. The results obtained for the differentiation pattern and protein production of porcine fibroblasts after irradiation with X-rays and high-energy carbon ions are in agreement with those obtained with human fibroblasts. An accumulation of fibrocytes with a concomitant increase in ECM protein production could be seen after in vitro irradiation. There is no indication of a higher RBE for fibrosis-related parameters compared with other endpoints (survival, chromosomal and DNA damage). The dose- and LET-dependence suggest that premature differentiation is a survival strategy after radiation damage. PMID- 11403712 TI - Release of mediators by DMSO-differentiated HL-60 cells exposed to electric interferential current and the requirement of biochemical prestimulation. AB - PURPOSE: Controversial results have been obtained with respect to the influence of biochemical prestimulation upon the action of electromagnetic fields on mammalian cells. Examined in detail has been the influence of a modulated low frequency (interferential) current upon the release of four mediators by DMSO differentiated HL-60 cells with and without prestimulation by the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP). MATERIALS AND METHODS: Human promyelocytes (HL-60) cells differentiated to granulocytes by dimethylsulphoxide were exposed to a so-called interferential current (IFC) in a suspension culture. The influence of IFC treatment, an amplitude-modulated 4000 Hz current used in therapeutic applications, on the release of prostaglandine E2 (PGE2), leukotriene B4 (LTB4), thromboxane B2 (TXB2) and interleukin 8 (IL-8) was investigated. Effects of a static magnetic field were also studied. RESULTS: In no case were significant differences between exposed and sham-exposed samples observed after 15 min exposure to selected modulation frequencies (0-125Hz, 250 microA cm(-2)). Therefore, the influence of prestimulation with different concentrations of fMLP on the release of the four mediators was investigated. fMLP alone enhanced the release of PGE2, LTB4, TXB2 and IL-8 with half of the maximum (ED50) effective concentration of approximately 10nM. Prestimulation with various concentrations of fMLP and subsequent exposure to IFC 125Hz (15 min, 250 microA cm(-2)) had no effect at low or high concentrations of fMLP, while in the middle range of concentration of approximately 100 pM fMLP a significant enhancement of PGE2, LTB4 and IL-8 was observed. This synergistic effect of fMLP and IFC was examined in detail in prestimulated cells (100pM fMLP) by varying the exposure time, current density and modulation frequency in the range 0-50 Hz. Window effects for the release of PGE2, LTB4 and IL-8 were obtained for exposure time, current density and modulation frequency whereas, in the release of TXB2, no field effect was detected. CONCLUSION: The study has demonstrated the synergistic action of chemical stimulation and electric current exposure, and the existence of windows for cellular activation status, exposure time, current density and modulation frequency. PMID- 11403713 TI - Neuropsychological sequelae of 50 Hz magnetic fields. AB - PURPOSE: The effects of occupational levels of 50 Hz magnetic fields on cognitive function were studied on 30 human volunteers. MATERIALS AND METHODS: The exposure system consisted of Merrit-type modified Helmholtz coils forming a 2 m cube, employing two orthogonal sets of coils producing a 28 microT (resultant) circularly polarized 50 Hz magnetic field. Subjects sat at a desk within the coils where they undertook a series of verbal and written tests of cognitive function. After these tests were concluded (approximately 30 min) subjects were either exposed or sham-exposed to fields (double-blinded) for 50 min. A second set of tests (employing alternate versions) was administered 20 min from the start of this period. Each subject returned after 7 days to repeat the sequence, but with the opposite field/sham status. RESULTS AND CONCLUSIONS: The majority of the results indicated no significant effect of exposure on cognition. However, verbal recall of a list of 15 words after an interference (the different list) was significantly impaired in the 'field on' condition. This is indicative of a reduction in short-term memory of words. The mean baseline score was somewhat higher in the 'field on' condition. However, this difference is within the parameters of normal variation. The other significant result occurred during performance of a trail-making task (alternating 1-A-2-B-3-C, etc.), with a decrease in performance as a result of exposure to the 50 Hz field. This task requires executive functioning with a working memory load, involving parietal spatial processing coordinated by prefrontal executive processing. In summary, the data are suggestive of detrimental effects on cognitive processes, particularly short-term learning and executive functioning. However, larger sample sizes are required to demonstrate statistically a more specific pattern of cognitive effects. PMID- 11403716 TI - Differentiation stages of eosinophils characterized by hyaluronic acid binding via CD44 and responsiveness to stimuli. AB - To characterize interleukin (IL)-5-induced eosinophils, we examined the expression of CD44, very late antigen (VLA)-4, and the IL-5 receptor alpha chain, as well as the levels of eosinophil peroxidase and the generation of superoxide. Eosinophils were prepared from IL-5-transgenic mice, then characterized using electron microscopy to determine their responses to stimuli. Whereas CD44 densities remained almost constant, the level of VLA-4 increased in parallel with eosinophil maturation. Although a subset of IL-5-induced eosinophils with high side scatter recovered from bone marrow and rare ones found in blood recognized hyaluronic acid (HA), most did not have this property. Bone marrow eosinophils with high side scatter and lower density contained eosinophil peroxidase, not only in granules, but also in membranous structures for 30% of this population. This population developed HA-binding ability in response to IL-3, IL-4, IL-5, granulocyte-macrophage colony-stimulating factor, macrophage inflammatory protein (MIP)-2, monocyte chemotactic protein (MCP)-1, eotaxin, nerve growth factor (NGF), and opsonized zymosan (OZ). Peripheral blood eosinophils acquired HA binding ability in response to the same stimuli, but their responses were less than those of bone marrow eosinophils with high levels of side scatter. However, splenic eosinophils did not respond to these stimuli. Although peripheral blood eosinophils did not proliferate when stimulated by IL-5, these were the only cells that released eosinophil peroxidase in response to IL-4, MIP-2, MCP-1, eotaxin, NGF, and OZ. With the exception of a subset of bone marrow eosinophils, the ability to acquire HA binding, but not the ability to generate superoxide, correlated with eosinophil peroxidase activity and major basic protein accumulation in the granules of maturing cells. PMID- 11403717 TI - Degenerate PCR-based cloning method for Eph receptors and analysis of their expression in the developing murine central nervous system and vasculature. AB - Eph receptors and their membrane-associated ephrin ligands regulate cell-cell interactions during development. The biochemical and biologic functions of this receptor tyrosine kinase family are still being elucidated but include roles in nervous system segmentation, axon pathfinding, and angiogenesis. To isolate murine orthologs of three zebrafish Eph family members (zek1, zek2, and zek3), we have used a degenerate RT-PCR-based cloning method specific for members of the Eph family. Although this method was effective for isolation of Eph receptor cDNAs, including members of both the A and B subfamilies, our results suggested that zek1 may not have a murine ortholog. The isolated cDNAs were also used to generate RNA in situ hybridization probes to examine the expression patterns of murine EphA2, A3, A4, A7, B1, B2, and B4 in 9.5-dpc mouse embryos. In addition to the expected abundant expression of these Eph receptors in the developing CNS and the presence of EphB receptors in vascular tissues, several of the EphA receptors were expressed in discrete regions of the developing vasculature. These results suggest a role for both EphA and EphB receptors in vascular development. PMID- 11403718 TI - Intraspleen DNA inoculation elicits protective cellular immune responses. AB - DNA immunization or inoculation is a recent vaccination method that induces both humoral and cellular immune responses in a range of hosts. Independent of the route or site of vaccination, the transfer of antigen-presenting cells (APC) or antigens into lymphoid organs is necessary. The aim of this investigation was to test whether intraspleen (i.s.) DNA inoculation is capable of inducing a protective immune response. We immunized mice by a single i.s. injection of a DNA construct expressing the immunoglobulin (Ig) heavy-chain variable domain (VH) in which the complementarity-determining regions (CDR) had been replaced by a Taenia crassiceps T-cell epitope. In these mice, immune responses and protective effects elicited by the vaccine were measured. We have shown here for the first time that i.s. DNA inoculation can induce protective cellular immune responses and activate CD8(+) T cells. Also, Ig V(H) appeared to be the minimal delivery unit of "antigenized" Ig capable of inducing T-cell activation in a lymphoid organ. The strategy of introducing T-cell epitopes into the molecular context of the V(H) domain in combination with i.s. DNA immunization could have important implications and applications for human immunotherapy. PMID- 11403720 TI - Significantly increased cortisol secretion in normal adrenocortical cells transfected with K-ras mutants derived from human functional adrenocortical tumors. AB - Our previous study showed that the mutation hotspots of the K-ras proto-oncogene in human functional adrenocortical tumors are in codons 15, 16, 18, and 31, thus differing from the sites in other tumors. In addition, analyzing the K-Ras protein by a recombinant DNA technique showed that the activity of endogenic GTPase and the GTPase-activating protein (GAP)-binding ability were significantly decreased in patients with these tumors. The aim of this study was to understand whether those K-ras mutants, which were found only in human adrenocortical tumors, play an important role in these tumors. Thus, the mutant K-ras cDNA was constructed with mammalian expression vectors and transfected into normal adrenocortical cells. The amount of cortisol secreted by the transfected cells was 20 to 30 times that of normal cells. Furthermore, Northern blot analysis revealed that the expression of the three steroidogenesis-related genes P450(scc) (cholesterol side-chain cleavage enzyme), P450(C17) (17alpha-hydroxylase/17, 20 lyase), and P450(C21) (steroid 21-hydroxylase) gene increased in the transfected cells. The K-ras oncogene significantly increases cortisol secretion by normal adrenocortical cells. PMID- 11403719 TI - Grb-2-associated binder-1 is involved in insulin-induced egr-1 gene expression through its phosphatidylinositol 3'-kinase binding site. AB - The Grb2-associated binder-1 (Gab1) is one of the major adapter molecules downstream of growth factor receptor signaling. Even though insulin causes tyrosine phosphorylation of Gab1, its role in insulin signaling has not been identified yet. We have demonstrated that insulin increased expression of early growth response gene-1 (egr-1), which is one of the most important transcription factors involved in cell proliferation and differentiation. In the present study, the possible role of Gab1 in insulin-induced egr-1 expression was studied using Rat1 fibroblasts expressing human insulin receptors and wildtype Gab1 (HIRc/Gab1(WT)), Gab1 with three tyrosines in the phosphatidylinositol (PI) 3' kinase binding domain mutated to phenylalanine (HIRc/Gab1(DeltaPI3K)), or histidinol resistance only (HIRc/HIS). Insulin-induced egr-1 expression in HIRc/Gab1(DeltaPI3K) cells was much lower than in the other cells, as determined by Northern blot analysis. These results suggest that Gab1 is involved in the signaling pathway for insulin-induced egr-1 expression through increasing PI3' kinase activity. The MAP kinase activity increased less with insulin treatment in HIRc/Gab1(DeltaPI3K) cells than in other cells. Inhibition of MAP kinase by the MEK inhibitor completely abolished insulin-induced egr-1 expression. These results suggest that Gab1 increases MAP kinase activity through its PI3'-kinase binding site, which then leads to egr-1 expression. Our results indicate that Gab1 is involved in the control of egr-1 expression regulated by insulin. PMID- 11403721 TI - Comparative analysis of the genetic structure and chromosomal location of the murine MyD118 (Gadd45beta) gene. AB - The MyD118 (Gadd45beta) protein is a member of a family of structurally related proteins, including Gadd45 (Gadd45alpha) and CR6 (Gadd45gamma), that have critical roles in regulating growth arrest and apoptosis. The MyD118 and other members of its family display distinct patterns of expression in response to stimuli that induce differentiation, growth arrest, or apoptosis. Species-blot analysis showed that MyD118 is an evolutionarily conserved gene, and comparative sequence analysis showed that MyD118 has a gene structure similar to that of other members of its gene family. Comparison of putative transcription factor binding sites found in sequences of this gene family provides evidence that p53 is involved in regulating the expression of MyD118 and that NF-kappaB may play a role in differential expression of MyD118 and Gadd45(Gadd45alpha). Fluorescence in situ hybridization localized the MyD118 gene to mouse chromosome band 10B5.3, correcting a previous assignment to mouse chromosome 9. PMID- 11403722 TI - What's new in Nicotine & Tobacco Research? PMID- 11403723 TI - Individual differences in smoking reward from de-nicotinized cigarettes. AB - Most studies of cigarette smoking and smoking cessation have focused on the psychopharmacological effects of nicotine; relatively few have explored the role of sensory aspects of cigarette smoke. Sensory aspects of cigarette smoke play a role in the maintenance of smoking behavior, and may be particularly important for certain smokers. This paper presents the results of a pooled analysis of nine studies conducted in our laboratory, in order to explore the influence of demographic and smoking-related variables on ratings of de-nicotinized as compared to nicotine-containing cigarettes. A major finding of this analysis is that ratings of smoking derived from de-nicotinized, but not nicotine-containing, cigarettes appear to vary with level of tobacco dependence, suggesting that sensory factors may be more important to highly dependent, as compared to less dependent, smokers. The implications of these findings for smoking cessation treatment and for future research are discussed. PMID- 11403724 TI - Withdrawal-suppressing effects of a novel smoking system: comparison with own brand, not own brand, and de-nicotinized cigarettes. AB - Tobacco smoking is associated with many health risks, all of which are decreased by smoking cessation. Tobacco companies are marketing novel products (e.g., Phillip Morris' Accord, R.J. Reynolds' Eclipse) intended to reduce these risks. Evaluation of these products is necessary to determine if they increase, decrease, or fail to change smoking's health risks. This study examines the acute effects of the Accord system on cigarette smokers; the study extends a previous preliminary evaluation by controlling for the system's eight-puff/cigarette limit, smokers' brand preference, and the act of smoking. Smokers of light or ultra-light cigarette brands (> or = 10 cigarettes/day, 16 men and 16 women) participated in this within-subjects protocol. Prior to each of the four, Latin square ordered, approximately 2.5-h sessions, subjects abstained from smoking for at least 8 h. In each session, subjects smoked eight puffs, at 30-min intervals, from either their own brand of cigarettes, a marketed brand that was not their own, de-nicotinized tobacco cigarettes, or the novel smoking system. Subjective and physiological effects were assessed regularly. Results confirmed previous observations that the withdrawal suppression, CO intake, and tachycardia produced by the novel system are all less than that produced by normally marketed cigarette brands, even controlling for the system's eight-puff limitation and for brand preference. Incomplete withdrawal suppression may increase smoking frequency, thus potentially offsetting any decreased risks associated with Accord use. Laboratory studies of the acute effects of novel smoking systems are an important part of a comprehensive evaluation program. PMID- 11403725 TI - Measurement of nicotine intake in pregnant women--associations to changes in blood cell count. AB - Self-reported information about smoking is imprecise and subject to bias, with accuracy varying according to circumstances. Biochemical assessment gives much clearer indications of the effects of tobacco intake on physiological parameters. As part of a randomized controlled trial, a new point-of-care test for smoking was used as a tool to reduce smoking in pregnancy. Measurements of nicotine metabolites in urine were related to a physiological effect of smoking, notably changes to blood parameters, assessed as a routine part of antenatal care. One hundred and eighty-seven pregnant women attending outpatient antenatal care were initially questioned by a midwife about smoking habits and later questioned by the investigators, during which the test was performed and the results relayed back to the patient. Self-reported smoking habit and cigarette consumption, either reported to midwives or the investigators, were shown to be poor indicators of the effects of smoking on blood parameters. The biochemical assessment of nicotine intake was significantly related to white blood count, haemoglobin concentration, haematocrit, mean cell volume, and mean cell haemoglobin. Red cell count, mean cell haemoglobin concentration, and platelet count were unrelated to nicotine metabolite measurements. We concluded that the new test was a reliable measure of nicotine intake in pregnancy and the results correlated with smoking-related changes to haematological parameters. PMID- 11403726 TI - Effects of a single dose of baclofen on self-reported subjective effects and tobacco smoking. AB - Baclofen has been reported in uncontrolled clinical studies to reduce craving for abused drugs and reduce their rewarding effects. The objective of the present study was to measure the acute effects of a single dose of baclofen on cigarette smoking, craving for nicotine, cigarette taste, and smoking satisfaction. Tobacco smokers (n = 16) who were not trying to quit received baclofen (20 mg) or placebo after overnight abstinence during two laboratory sessions in a within-subjects design. We measured the subjective effects of baclofen on mood and self-reported ratings of craving for nicotine, and on the number of cigarettes smoked of the subjects' preferred brand during a 3-h ad libitum smoking period. Baclofen did not change the number of cigarettes smoked by the subjects nor did it change ratings of nicotine craving. However, baclofen altered the sensory properties of smoked cigarettes (e.g., increasing ratings of 'harsh' and decreasing ratings of 'like cigarette's effects'). It also produced mild sedative-like subjective effects, such as increases in feeling 'relaxed'. Thus, although baclofen did not reduce cigarette craving or smoking in the present study, it did produce some mood-altering effects and changes in sensory aspects of smoking that may facilitate smoking cessation. PMID- 11403727 TI - Relationship between drug exposure and the efficacy and safety of bupropion sustained release for smoking cessation. AB - A population pharmacokinetic and pharmacodynamic analysis evaluated the relationships of dose, plasma concentrations of bupropion and metabolites, and patient covariates with the safety and efficacy of bupropion sustained release (SR) for smoking cessation. A total of 519 outpatient chronic cigarette smokers were randomized to one of three bupropion SR doses: 100, 150, or 300 mg/day or placebo. The bupropion plasma concentration time data were fit and subject specific bayesian estimates of clearance were obtained. Logistic regression analyses evaluated the role of dose, concentrations, and covariates in predicting efficacy and safety endpoints. For the evaluation of efficacy, patients were classified as quitters or non-quitters on the basis of a 4-week quit variable (defined as complete abstinence for weeks 4-7 of the study). For the evaluation of safety, patients were classified into two categories for each adverse event evaluated, corresponding to whether the patient ever experienced the adverse event during the course of the study or never experienced the event, regardless of whether the event was treatment-emergent. The efficacy of bupropion SR in facilitating smoking cessation was found to be related to dose and a mean metabolite concentration, and quitting in general was found to be related to the number of cigarettes smoked per day at baseline. Smoking cessation was 1.42, 1.69, and 2.84 times more likely in patients receiving 100, 150, and 300 mg/day of bupropion SR, respectively, as compared to placebo (p = 0.0001). As the baseline number of cigarettes smoked per day increased, the likelihood of quitting decreased regardless of the treatment condition. Insomnia and dry mouth were positively associated with mean metabolite concentrations, and dry mouth was inversely related to patient weight. Anxiety was inversely related to predicted steady-state concentration (Cpss), suggesting a positive effect on this withdrawal symptom. Bupropion SR exhibits a statistically significant dose/plasma level-response relationship for smoking cessation. Dry mouth and insomnia, related to concentrations, may be managed with dose reduction, with the realization that smoking cessation may be impaired. PMID- 11403728 TI - Sex differences in the subjective and reinforcing effects of visual and olfactory cigarette smoke stimuli. AB - Although nicotine intake clearly reinforces cigarette smoking behavior, non nicotine smoke stimuli may become conditioned reinforcers of smoking. In Study 1, we compared the acute subjective and reinforcing effects of cigarette smoking in men and women under two conditions: blockade of visual and olfactory/taste smoke stimuli vs. no blockade. Subjective hedonic ratings of 'like puffs' and 'satisfying', but not 'strength', 'high in nicotine', or CO boost, were significantly reduced under the blockade vs. no blockade conditions. During subsequent ad lib puffing, significantly fewer puffs were self-administered under the blockade condition, particularly among women. In Study 2, we examined the influences of these stimuli separately and found that olfactory/taste stimuli, but not visual stimuli, reduced hedonic ratings and puff self-administration in women but not in men. In Study 3, procedures similar to those in Study 1 were used to examine whether this sex difference in responses to conditioned stimuli generalizes to a non-drug consummatory behavior, eating (pizza). However, hedonic ratings and ad lib consumption of pizza were substantially reduced in both men and women following blockade of visual and olfactory/taste food stimuli. These results indicate that the presumably conditioned stimuli of olfactory/taste from cigarette smoke may influence subjective hedonic ratings and reinforcement from smoking more in women than in men. However, this sex difference may not generalize beyond smoking or other drug reinforcement. PMID- 11403729 TI - Nicotine dependence, withdrawal symptoms, and adolescents' readiness to quit smoking. AB - The purpose of this study was to characterize nicotine dependence and nicotine withdrawal symptoms among adolescent smokers and to investigate associations between these key factors and adolescents' readiness to quit smoking. A total of 5624 high school students participated in a school-based survey. Of 1111 adolescents who were current or former smokers, the following stage-of-change distribution for smoking cessation was observed: precontemplation, 52.5%; contemplation, 16.0%; preparation, 7.5%; action, 13.2%; and maintenance, 10.8%. Among current smokers, 18.1% were substantially dependent on nicotine, 45.2% had moderate dependence, and 36.7% had no dependence. Higher proportions of current smokers than successful quitters reported withdrawal symptoms with their most recent quit attempts. Precontemplators exhibited significantly higher mean nicotine dependence scores than did students in the contemplation or preparation stages (F(2,837) = 12.03; p < 0.0001). A similar trend was observed for withdrawal-symptom scores across the stages of change. The nicotine dependence and withdrawal-symptom scores were significantly correlated (r = 0.44, p < 0.001). Nicotine dependence and nicotine withdrawal appear to interfere with adolescents' abilities and readiness to quit smoking, suggesting a potential role for nicotine replacement therapy in the treatment of tobacco use and dependence among adolescents. PMID- 11403730 TI - Tolerability of concurrent use of nicotine gum and smoking in healthy volunteers. AB - Although nicotine replacement therapy (NRT) is mainly licensed as an aid for smoking cessation, many smokers alternate NRT with cigarettes. This study evaluated the cardiovascular tolerability of nicotine gum plus smoking. This open, three-way, randomized, multiple-dose, crossover study involved 19 healthy adult volunteer smokers. Three treatments (placebo gum+cigarette vs. nicotine 4 mg gum+unlit cigarette vs. nicotine 4-mg gum+cigarette) were each administered hourly seven times during one study day. Plasma nicotine, ECG variables (ST60 and ST(slope)), heart rate, and blood pressure were measured at rest and during/after 5 min exercise tests. Mean plasma nicotine levels were higher with gum plus smoking than with either gum or smoking alone. After the final administration, mean levels with gum plus smoking were 39.5 (range 27.6-54.4) ng/ml vs. 19.6 (11.8-30.1) ng/ml with gum and 22.6 (6.6-36.6) ng/ml with smoking alone. No signs of myocardial ischaemia were observed during concomitant use of gum plus smoking, despite high plasma nicotine levels. Decreases of ST60 and ST(slope) were observed in all treatment groups during exercise but were asymptomatic. Concomitant gum plus smoking did not increase the incidence of extrasystoles or arrhythmia. No serious adverse events occurred during the study. In conclusion, there were no signs of myocardial ischaemia during multiple submaximal exercise tests in healthy volunteers while smoking and using nicotine 4-mg gum. PMID- 11403731 TI - Ethnic variation in peer influences on adolescent smoking. AB - Previous research has indicated that the influence of peers on adolescent smoking may differ across ethnic groups. Although many studies have focused on African Americans, Hispanics, and Whites, few studies have included Asian Americans, Pacific Islanders, and multi-ethnic adolescents as distinct groups. Using data from a statewide sample of 5870 eighth-grade adolescents in California, this study examined ethnic differences in the association between peer influence variables and smoking behavior and susceptibility. Informational peer influence (best friends' smoking behavior) and normative peer influence (prevalence estimates of peer smoking) were investigated. We hypothesized that informational peer influences would be stronger among Whites (whose families originate primarily from the individualistic cultures of the USA and Western Europe) than among Asian Americans, Pacific Islanders, Hispanics, and African Americans (whose families originate primarily from collectivist cultures). Conversely, we hypothesized that normative peer influences would be stronger among ethnic minority adolescents from collectivist cultural backgrounds than among Whites. Consistent with previous studies, friends' smoking and prevalence estimates of peer smoking were risk factors for past 30-day smoking and susceptibility to smoking across ethnic groups. The influence of friends' smoking behavior was stronger among Whites than among several other groups: Pacific Islanders, African Americans, and Hispanic/Latinos. The influence of prevalence estimates of peer smoking was stronger among Whites than among multiethnic adolescents. Results indicate that cultural factors may play a role in peer influences on smoking initiation. Smoking prevention interventions for adolescents should address the differences in peer influences across ethnic groups. PMID- 11403732 TI - Cotinine binding to nicotinic acetylcholine receptors in bovine chromaffin cell and rat brain membranes. AB - Cotinine is the major metabolite of nicotine. It has nicotine-like biological activity, but its potency is low. We studied cotinine binding to nicotinic receptors labelled with [3H]epibatidine. In membranes from cultured bovine chromaffin cells [3H]epibatidine bound to two apparent sites with K(d) values of 93 and 1400 pM. The low-affinity binding represented two-thirds of the binding sites. In rat frontal cortex and hippocampus homogenate membranes, only one apparent binding site was detected. The Kd values were 40 and 62 pM, in frontal cortex and hippocampus, respectively. Nicotine displaced [3H]epibatidine 10 times more potently from the brain than from the chromaffin cell membranes, and cotinine had over two orders of magnitude lower affinity than nicotine. In addition, the competitive nicotinic receptor antagonists methyllycaconitine and dihydro beta-erythroidine displaced [3H]epibatidine (100 pM and 1 nM) from the chromaffin cell membranes. Alpha-bungarotoxin did not affect the binding of 100 pM [3H]epibatidine. However, upon labelling with 1 nM [3H]epibatidine alpha bungarotoxin (10 nM to 10 microM) displaced one-sixth of the bound radioligand. Our results demonstrate that 100 pM to 1 nM [3H]epibatidine labels mostly neuronal heteropentameric nicotinic receptors in bovine chromaffin cell membranes, and that cotinine is a low-affinity nicotinic ligand both in the adrenal chromaffin cell and in the brain receptors. PMID- 11403734 TI - Quality-of-life, legal-financial, and disability issues in fibromyalgia. AB - Patients with fibromyalgia have an altered quality of life that is hard to quantitate using existing indices. The principal legal issues associated with the syndrome are: Does fibromyalgia exist? Can it be caused by or flared by stress or trauma? Does disability apply to fibromyalgia and if so, how? These issues are critically reviewed. PMID- 11403735 TI - Classification, epidemiology, and natural history of fibromyalgia. AB - Fibromyalgia (FM), also known as fibromyalgia syndrome (FMS) and fibrositis, is a common form of nonarticular rheumatism that is associated with chronic generalized musculoskeletal pain, fatigue, and a long list of other complaints. Some have criticized the classification of FM as a distinct medical entity, but existing data suggest that individuals meeting the case definition for FM are clinically somewhat distinct from those with chronic widespread pain who do not meet the full FM definition. Clinic studies have found FM to be common in countries worldwide; these include studies in specialty and general clinics. The same is true of general population studies, which show the prevalence of FM to be between 0.5% and 5%. Knowledge about risk factors for FM is limited. Females are at greater risk, and risk appears to increase through middle age, then decline. Although some authors claim that an epidemic of FM has been fueled by an over generous Western compensation system, there are no data that demonstrate an increasing incidence or prevalence of FM; moreover, existing data refute any association between FM prevalence and compensation. Claims that the FM label itself causes illness behavior and increased dependence on the medical system also are not supported by existing research. This article reviews the classification, epidemiology, and natural history of FM. PMID- 11403736 TI - Psychosocial aspects of fibromyalgia. AB - The view that fibromyalgia syndrome (FMS) is a psychiatric disorder or can be caused by stress or abuse is unproven. The construct of posttraumatic FMS has not been adequately validated. Similarly, there is no evidence that communicating the diagnosis to patients causes iatrogenic consequences. Research suggesting a higher rate of posttraumatic stress disorder among those with FMS is weak. More research examining specific psychological processes in FMS is desirable. Because of the potential for harm to patients, clinicians should be cognizant of possible undue influences on medical opinion by agencies providing health care and research funding. PMID- 11403737 TI - Nociceptive aspects of fibromyalgia. AB - Although characterized by a variety of symptoms, chronic widespread pain is the primary complaint bringing most patients with fibromyalgia syndrome (FMS) into the clinic. The etiology of this painful condition is unknown, and any possible relationship between pain and the many other symptoms of FMS is unclear. This article focuses on the unique characteristics of nociception in patients with FMS. The intent is to present criteria that should be considered in the search for biological events that contribute to FMS pain. Based on this approach, examples are proposed of factors that fulfill some criteria and may, therefore, deserve further study for their possible role in pain associated with FMS. PMID- 11403738 TI - Overlap of fibromyalgia with other medical conditions. AB - Fibromyalgia is a multisystem illness. One of its defining features, generalized pain, may also be present in other rheumatic entities. The diagnosis of fibromyalgia is not easy by any means, it requires a profound knowledge of internal medicine. This article discusses the different rheumatic and nonrheumatic diseases that overlap or are prone to be confused with fibromyalgia. It emphasizes the key points in the differential diagnosis. PMID- 11403740 TI - Cervicogenic headache: diagnostic evaluation and treatment strategies. AB - Cervicogenic headache is a chronic, hemicranial pain syndrome in which the source of pain is located in the cervical spine or soft tissues of the neck but the sensation of pain is referred to the head. The trigeminocervical nucleus is a region of the upper cervical spinal cord where sensory nerve fibers in the descending tract of the trigeminal nerve converge with sensory fibers from the upper cervical roots. This convergence of upper cervical and trigeminal nociceptive pathways allows the referral of pain signals from the neck to the trigeminal sensory receptive fields of the face and head. The clinical presentation of cervicogenic headache suggests that there is an activation of the trigeminovascular neuroinflammatory cascade, which is thought to be one of the important pathophysiologic mechanisms of migraine. Another convergence of sensorimotor fibers has been described involving intercommunication between the spinal accessory nerve (CN XI), the upper cervical nerve roots, and ultimately the descending tract of the trigeminal nerve. This neural network may be the basis for the well- recognized patterns of referred pain from the trapezius and sternocleidomastoid muscles to the face and head. Diagnostic criteria have been established for cervicogenic headache but its presenting characteristics may be difficult to distinguish from migraine, tension-type headache, or hemicrania continua. A multidisciplinary treatment program integrating pharmacologic, nonpharmacologic, anesthetic, and rehabilitative interventions is recommended. This article reviews the clinical presentation of cervicogenic headache, its diagnostic evaluation, and treatment strategies. PMID- 11403739 TI - Pharmacologic treatment of fibromyalgia. AB - Fibromyalgia is a chronic syndrome characterized by widespread pain, unrefreshed sleep, disturbed mood, and fatigue. Until such time as we have a clearer understanding of the trigger and/or pathophysiologic mechanisms producing these symptoms, pharmacologic treatment should be aimed at individual symptoms. Such treatment should ideally be offered as part of a multidisciplinary treatment program using both pharmacologic and nonpharmacologic treatment modalities. Critical components of any successful fibromyalgia treatment program include addressing physical fitness, work and other functional activities, and mental health, in addition to symptom-specific therapies. The main symptoms that should be addressed include pain, sleep disturbances including restless leg syndrome, mood disturbances, and fatigue. Pharmacologic therapy should also be considered for syndromes commonly associated with fibromyalgia including irritable bowel syndrome, interstitial cystitis, migraine headaches, temporomandibular joint dysfunction, dysequilibrium including neurally mediated hypotension, sicca syndrome, and growth hormone deficiency. This article provides general guidelines in initiating a successful pharmacologic treatment program for fibromyalgia. PMID- 11403741 TI - Cervicogenic headache: manual and manipulative therapies. AB - This article reviews current literature on the role of manual medicine in the diagnosis and treatment of cervicogenic headache. Manual diagnostic procedures and treatment procedures are described for the cervical spine. Emphasis is placed on accurate diagnosis using a biomechanical model and precise localization of forces. Muscle energy technique is suggested as a safe and effective treatment when somatic dysfunction of the cervical spine is found in association with the diagnostic criteria for cervicogenic headache. Lastly, a suggested clinical approach to this problem from a manual medicine perspective is given. PMID- 11403742 TI - Regional myofascial pain syndrome and headache: principles of diagnosis and management. AB - Myofascial pain is frequently overlooked in dealing with headache pain. Myofascial pain is defined as pain and/or autonomic phenomena referred from active trigger points, with associated dysfunction. The trigger point is a focus of hyperirritability in the muscle, that when compressed, is locally tender, and if sensitized, gives rise to referred pain and tenderness. The therapy for myofascial pain requires enhancing central inhibition through pharmacology or behavioral techniques and simultaneously reducing peripheral inputs through physical therapies including exercises and trigger point-specific therapy. PMID- 11403743 TI - Cervicogenic headache: anatomic basis and pathophysiologic mechanisms. AB - Cervicogenic headache is pain perceived in the head but referred from a primary source in the cervical spine. The physiologic basis for this pain is convergence between trigeminal afferents and afferents from the upper three cervical spinal nerves. The possible sources of cervicogenic headache lie in the structures innervated by the C1 to C3 spinal nerves, and include the upper cervical synovial joints, the upper cervical muscles, the C2-3 disc, the vertebral and internal carotid arteries, and the dura mater of the upper spinal cord and posterior cranial fossa. Experiments in normal volunteers have established that the cervical muscles and joints can be sources of headache. PMID- 11403744 TI - Cervicogenic headache: clinical presentation, diagnostic criteria, and differential diagnosis. AB - Since the first attempt at setting down diagnostic criteria was made in 1990, there has been considerable progress in the field of cervicogenic headache (CEH). CEH makes up a "final common pathway" for several neck disorders that may originate at different levels of the cervical spine. CEH has been defined as being mainly a unilateral headache without sideshift; it may accordingly also be bilateral. Anesthetic blockades are mandatory for scientific work. If the pain is bilateral, it is particularly important that blockades are carried out. Pain stemming from the neck usually spreads to the oculofrontotemporal area. The most characteristic features are symptoms and signs of neck involvement (such as mechanical precipitation of attack, and so forth). Migraine without aura and tension-type headache are the most difficult differential diagnosis problems. PMID- 11403745 TI - The distribution of the effects of genes affecting quantitative traits in livestock. AB - Meta-analysis of information from quantitative trait loci (QTL) mapping experiments was used to derive distributions of the effects of genes affecting quantitative traits. The two limitations of such information, that QTL effects as reported include experimental error, and that mapping experiments can only detect QTL above a certain size, were accounted for. Data from pig and dairy mapping experiments were used. Gamma distributions of QTL effects were fitted with maximum likelihood. The derived distributions were moderately leptokurtic, consistent with many genes of small effect and few of large effect. Seventeen percent and 35% of the leading QTL explained 90% of the genetic variance for the dairy and pig distributions respectively. The number of segregating genes affecting a quantitative trait in dairy populations was predicted assuming genes affecting a quantitative trait were neutral with respect to fitness. Between 50 and 100 genes were predicted, depending on the effective population size assumed. As data for the analysis included no QTL of small effect, the ability to estimate the number of QTL of small effect must inevitably be weak. It may be that there are more QTL of small effect than predicted by our gamma distributions. Nevertheless, the distributions have important implications for QTL mapping experiments and Marker Assisted Selection (MAS). Powerful mapping experiments, able to detect QTL of 0.1sigma(p), will be required to detect enough QTL to explain 90% the genetic variance for a quantitative trait. PMID- 11403746 TI - Power analysis of QTL detection in half-sib families using selective DNA pooling. AB - Individual loci of economic importance (QTL) can be detected by comparing the inheritance of a trait and the inheritance of loci with alleles readily identifiable by laboratory methods (genetic markers). Data on allele segregation at the individual level are costly and alternatives have been proposed that make use of allele frequencies among progeny, rather than individual genotypes. Among the factors that may affect the power of the set up, the most important are those intrinsic to the QTL: the additive effect of the QTL, and its dominance, and distance between markers and QTL. Other factors are relative to the choice of animals and markers, such as the frequency of the QTL and marker alleles among dams and sires. Data collection may affect the detection power through the size of half-sib families, selection rate within families, and the technical error incurred when estimating genetic frequencies. We present results for a sensitivity analysis for QTL detection using pools of DNA from selected half sibs. Simulations showed that conclusive detection may be achieved with families of at least 500 half-sibs if sires are chosen on the criteria that most of their marker alleles are either both missing, or one is fixed, among dams. PMID- 11403747 TI - Genetic components of litter size variability in sheep. AB - Classical selection for increasing prolificacy in sheep leads to a concomitant increase in its variability, even though the objective of the breeder is to maximise the frequency of an intermediate litter size rather than the frequency of high litter sizes. For instance, in the Lacaune sheep breed raised in semi intensive conditions, ewes lambing twins represent the economic optimum. Data for this breed, obtained from the national recording scheme, were analysed. Variance components were estimated in an infinitesimal model involving genes controlling the mean level as well as its environmental variability. Large heritability was found for the mean prolificacy, but a high potential for increasing the percentage of twins at lambing while reducing the environmental variability of prolificacy is also suspected. Quantification of the response to such a canalising selection was achieved. PMID- 11403748 TI - Genetic relationship between cyclic ovarian activity in heifers and cows and beef traits in males. AB - Records were collected in an experimental herd over an 11-year period from purebred Charolais heifers (n=351), cows (n=615) and young entire bulls (n=383). The objective of the study was to estimate the genetic relationship between the components of female ovarian activity (age at puberty and postpartum anoestrus length), their growth rate and body condition score and beef traits measured on related bulls. Two methods were used to estimate age at puberty and postpartum anoestrus length: the detection of oestrous behaviour and a test of cyclicity based on plasmatic progesterone assay. This study shows the existence of significant heritability estimates for the different cyclicity traits (h(2) between 0.11 and 0.38). Most of the genetic correlation coefficients between ovarian activity and growth rate of females and males are negative and favourable (r(g) between - 0.43 and 0.06). Cyclicity is also favourably related with body condition score in young or adult females (r(g) between - 0.65 and - 0.22). The genetic relationship between female ovarian activity and proportion of adipose tissue in the male carcass is, however, close to zero. These results show that an antagonism between male beef traits measured in this study and female ovarian activity is unlikely to be a cause for concern in the short term. PMID- 11403749 TI - Detection of quantitative trait loci for growth and fatness in pigs. AB - A quantitative trait locus (QTL) analysis of growth and fatness data from a three generation experimental cross between Meishan (MS) and Large White (LW) pig breeds is presented. Six boars and 23 F1 sows, the progeny of six LW boars and six MS sows, produced 530 F2 males and 573 F2 females. Nine growth traits, i.e. body weight at birth and at 3, 10, 13, 17 and 22 weeks of age, average daily gain from birth to 3 weeks, from 3 to 10 weeks and from 10 to 22 weeks of age, as well as backfat thickness at 13, 17 and 22 weeks of age and at 40 and 60 kg live weight were analysed. Animals were typed for a total of 137 markers covering the entire porcine genome. Analyses were performed using two interval mapping methods: a line-cross (LC) regression method where founder lines were assumed to be fixed for different QTL alleles and a half-/full-sib (HFS) maximum likelihood method where allele substitution effects were estimated within each half-/full sib family. Both methods revealed highly significant gene effects for growth on chromosomes 1, 4 and 7 and for backfat thickness on chromosomes 1, 4, 5, 7 and X, and significant gene effects on chromosome 6 for growth and backfat thickness. Suggestive QTLs were also revealed by both methods on chromosomes 2 and 3 for growth and 2 for backfat thickness. Significant gene effects were detected for growth on chromosomes 11, 13, 14, 16 and 18 and for backfat thickness on chromosome 8, 10, 13 and 14. LW alleles were associated with high growth rate and low backfat thickness, except for those of chromosome 7 and to a lesser extent early-growth alleles on chromosomes 1 and 2 and backfat thickness alleles on chromosome 6. PMID- 11403750 TI - Genetic diversity measures of local European beef cattle breeds for conservation purposes. AB - This study was undertaken to determine the genetic structure, evolutionary relationships, and the genetic diversity among 18 local cattle breeds from Spain, Portugal, and France using 16 microsatellites. Heterozygosities, estimates of Fst, genetic distances, multivariate and diversity analyses, and assignment tests were performed. Heterozygosities ranged from 0.54 in the Pirenaica breed to 0.72 in the Barrosa breed. Seven percent of the total genetic variability can be attributed to differences among breeds (mean F(st) = 0.07; P<0.01). Five different genetic distances were computed and compared with no correlation found to be significantly different from 0 between distances based on the effective size of the population and those which use the size of the alleles. The Weitzman recursive approach and a multivariate analysis were used to measure the contribution of the breeds diversity. The Weitzman approach suggests that the most important breeds to be preserved are those grouped into two clusters: the cluster formed by the Mirandesa and Alistana breeds and that of the Sayaguesa and Tudanca breeds. The hypothetical extinction of one of those clusters represents a 17% loss of diversity. A correspondence analysis not only distinguished four breed groups but also confirmed results of previous studies classifying the important breeds contributing to diversity. In addition, the variation between breeds was sufficiently high so as to allow individuals to be assigned to their breed of origin with a probability of 99% for simulated samples. PMID- 11403751 TI - Influence of diuretics on the concentration of proteins and other components of pleural transudates in patients with heart failure. AB - PURPOSE: Diuretic therapy increases the total protein and lactate dehydrogenase concentrations in pleural fluid in patients with transudates due to heart failure, but the effect of diuresis on other substances in pleural fluid constituents is not known. SUBJECTS AND METHODS: Twenty-one patients with transudative pleural effusions due to congestive heart failure were prospectively studied. Repeated diagnostic thoracentesis (mean +/- SD = 3 +/- 1; range, 2 to 6) was performed until the effusions were radiographically unapparent (5 +/- 2 days). Thirty-one patients with congestive heart failure who underwent only a single thoracentesis after diuretic therapy served as controls. We measured the concentrations of various components of pleural effusions in the serum and in the pleural fluid, and determined the serum-pleural fluid gradient (serum concentration minus pleural fluid concentration) and ratio (serum concentration divided by pleural fluid concentration). RESULTS: The pleural concentrations of most components increased significantly (P <0.001) from the initial specimen to the final specimen: total protein, from 23 +/- 7 g/L to 33 +/- 9 g/L; albumin, from 13 +/- 4 g/L to 18 +/- 6 g/L; lactate dehydrogenase, from 177 +/- 62 U/L to 288 +/- 90 U/L; cholesterol, from 31 +/- 16 mg/dL to 52 +/- 22 mg/dL; and cholinesterase, from 1,304 +/- 616 U/L to 1,884 +/- 674 U/L. Expressed as percentage change, the increases in the serum-pleural fluid gradients for albumin (12% +/- 22%) and total protein (11% +/- 12%) were significantly less than the increases in their concentrations in pleural fluid (albumin, 47% +/- 49%; total protein, 48% +/- 40%) or in their pleural fluid/serum ratios (albumin, 27% +/- 29%; total protein, 38% +/- 34%). CONCLUSIONS: The concentrations of the biochemical components commonly measured in pleural fluid increase progressively during diuretic therapy. Calculation of the serum-pleural fluid gradients for protein and albumin may be the most useful way to distinguish transudates from exudates in patients with congestive heart failure who have undergone diuresis. PMID- 11403752 TI - Correlation between 25-hydroxyvitamin D3 1 alpha-hydroxylase gene expression in alveolar macrophages and the activity of sarcoidosis. AB - PURPOSE: To demonstrate expression of the 25-hydroxyvitamin D3 1 alpha hydroxylase (1 alpha-hydroxylase) gene in human alveolar macrophages and measure the correlations among the 1 alpha-hydroxylase mRNA level, the activity of sarcoidosis, and calcium metabolism. SUBJECTS AND METHODS: We examined 7 patients with sarcoidosis and 6 control patients with other pulmonary disorders who underwent bronchoalveolar lavage. Levels of 1 alpha-hydroxylase mRNA were measured by semiquantitative polymerase chain reaction amplification. We measured serum levels of calcium, ionized calcium, parathyroid hormone, calcitriol (1,25 dihydroxyvitamin D3), and 25-hydroxyvitamin D3 to evaluate calcium metabolism. To estimate the activity of sarcoidosis, we measured the cell count, the CD4/CD8 ratio in bronchoalveolar lavage cells, and the serum angiotensin-converting enzyme (ACE) activity. RESULTS: Expression of 1 alpha-hydroxylase was demonstrated in purified human alveolar macrophages. The 1 alpha-hydroxylase mRNA levels in bronchoalveolar lavage cells were fivefold higher in sarcoidosis patients than in control patients (10.8 +/- 3.6 vs. 2.2 +/- 1.4, P <0.003). Among all patients studied, there were significant correlations between the 1 alpha hydroxylase mRNA level in bronchoalveolar lavage samples and the percentage of alveolar lymphocytes (r = 0.83, P <0.005), the CD4/CD8 ratio (r = 0.77, P <0.02), serum ACE level (r = 0.58, P <0.05), serum ionized calcium level (r = 0.58, P <0.05), and the calcitriol/25-hydroxyvitamin D3 ratio (r = 0.57, P <0.05). In the sarcoidosis patients, a significant correlation was also observed between 1 alpha hydroxylase mRNA and the percentage of alveolar lymphocytes (r = 0.82, P <0.05). CONCLUSION: There is a correlation between 1 alpha-hydroxylase gene expression in alveolar macrophages with the activity of sarcoidosis and its associated disturbances in calcium metabolism. PMID- 11403753 TI - Early cardioversion of atrial fibrillation facilitated by transesophageal echocardiography: short-term safety and impact on maintenance of sinus rhythm at 1 year. AB - BACKGROUND: For patients presenting with atrial fibrillation of only a few weeks duration, the use of transesophageal echocardiography offers the opportunity to markedly abbreviate the duration of atrial fibrillation before cardioversion. We sought to determine if the shorter duration of atrial fibrillation allowed by a transesophageal echocardiography strategy had an impact on the recurrence of atrial fibrillation and prevalence of sinus rhythm during the first year following cardioversion. METHODS: Transesophageal echocardiography was attempted in 539 patients (292 men, 247 women; 71.6 +/- 13.0 years.) with atrial fibrillation > or =2 days (66.1% <3 weeks) or of unknown duration before elective cardioversion of atrial fibrillation. Therapeutic anticoagulation at the time of transesophageal echocardiography was present in 94.6% of patients, and 73.4% of subjects were discharged on warfarin. RESULTS: Atrial thrombi were identified in 70 (13.1%) patients. Successful cardioversion in 413 patients without evidence of atrial thrombi was associated with clinical thromboembolism in 1 patient (0.24%, 95% confidence interval: 0.0--0.8%). In patients with atrial fibrillation <3 weeks at the time of cardioversion (a duration incompatible with conventional therapy of 3 to 4 weeks of warfarin before cardioversion), the 1-year atrial fibrillation recurrence rate was lower (41.1% vs. 57.9%, P <0.01), and the prevalence of sinus rhythm at 1 year was increased (65.8% vs. 51.3%, P <0.03). No other clinical or echocardiographic index was associated with recurrence of atrial fibrillation or sinus rhythm at 1 year. CONCLUSIONS: Early cardioversion facilitated by transesophageal echocardiography has a favorable safety profile and provides the associated benefit of reduced recurrence of atrial fibrillation for patients in whom the duration of atrial fibrillation is <3 weeks. PMID- 11403754 TI - The association between CagA status and the development of esophagitis after the eradication of Helicobacter pylori. AB - BACKGROUND: Strains of Helicobacter pylori with the cytotoxine-associated gene A (cagA) are linked to severe forms of gastroduodenal disease. Although eradication of H. pylori may predispose to the development of reflux esophagitis, the effects of CagA status on risk of esophagitis after successful H. pylori treatment are not known. METHODS: We studied 50 consecutive patients without esophagitis in whom H. pylori was eradicated successfully. CagA status was determined by immunoblotting sera from patients against H. pylori antigens. Patients underwent upper gastrointestinal endoscopy before eradication and 6, 12, 18, and 24 months after eradication or when reflux symptoms occurred. Biopsy specimens of the antrum and corpus were evaluated for gastritis before H. pylori eradication and at the end of the study. The sum of the scores for acute and chronic inflammation (both measured on a 0 [absent] to 3 [severe] scale) comprised the total gastritis severity score. RESULTS: In a multivariate proportional hazards regression analysis, positive CagA serology (hazard ratio [HR] = 10, 95% confidence interval [CI]: 1.3 to 81) and moderate-to-severe corpus gastritis (total severity score > or =4) before eradication (HR = 2.3, 95% CI: 1.2 to 6.1) were independent risk factors for the development of esophagitis after H. pylori eradication. CONCLUSION: Patients infected with strains of H. pylori that are cagA-positive are at increased risk of developing esophagitis after eradication of H. pylori. PMID- 11403755 TI - Stent-supported recanalization of chronic iliac artery occlusions. AB - PURPOSE: Iliac artery occlusions that are more than a few centimeters in length are normally treated with surgical bypass grafting. The aim of this study was to evaluate the results of primary stent implantation after Excimer laser-assisted recanalization of iliac artery occlusions. SUBJECTS AND METHODS: We studied 212 consecutive patients with chronic unilateral iliac artery occlusions (mean [+/- SD] length 8.9 +/- 3.9 cm) who were treated with Excimer laser-assisted recanalization and stent implantation. Based on the criteria of the Society of Cardiovascular and Interventional Radiology, lesions were graded as class III occlusions (<5 cm) in 46 patients and as class IV (> or =5 cm) in 166 patients. A total of 527 stents (Palmaz stent, 346; Wallstent, 94; Strecker stent, 38; covered stents, 49) were implanted. RESULTS: Technical success was achieved in 190 (90%) patients. There was a clinical improvement of three grades in 112 (53%) patients and of two grades in 67 (32%) patients. The rate of major complications was 1.4%, which included arterial rupture (1) and embolic events (2). Primary patency rates were 84% at 1 year, 81% at 2 years, 78% at 3 years, and 76% at 4 years. Secondary patency rates were 88% at 1 year, 88% at 2 years, 86% at 3 years, and 85% at 4 years. CONCLUSION: Stent-supported angioplasty is an effective treatment for iliac artery occlusions, with less morbidity and mortality than is associated with surgery. However, reported long-term patency rates after bypass surgery are greater than those we observed with interventional treatment. The value of primary stenting as compared with angioplasty alone should be evaluated in a randomized trial. PMID- 11403756 TI - Do statins cause cancer? A meta-analysis of large randomized clinical trials. AB - PURPOSE: Although the short-term safety and tolerability of statins has been well established, their potential carcinogenicity in the long term is still debated. The goal of this study was to determine whether long-term treatment with statins is associated with an increased risk of fatal and nonfatal cancers. METHODS: We searched the Medline database between January 1966 and December 1999 for randomized, controlled trials of human subjects in which monotherapy with a statin was compared with placebo. No language restrictions were applied. Only trials with a minimum treatment duration of 4 years and a minimum of 1,000 subjects were included. Studies that did not provide information on fatal or nonfatal cancers were excluded. Data on fatal and nonfatal cancers and all-cause mortality were extracted by a single nonblinded reviewer. Overall crude estimates of risk difference were computed by summing the numerators and denominators of trial-specific risk estimates. RESULTS: Five trials met the inclusion criteria. The estimated differences in absolute risk between treatment and placebo were as follows (negative risks indicate that treatment was safer than placebo): all nonfatal cancers, 0.0% (95% confidence interval [CI]: -0.8% to 0.8%); all fatal cancers, -0.1% (95% CI: -0.7% to 0.4%); all fatal and nonfatal cancers combined, 0.1% (95% CI: -1.0% to 0.7%); and all-cause mortality, -1.5% (95% CI: 2.8% to 0.2%). CONCLUSION: This study demonstrates no association between statin use over a 5-year period and the risk of fatal and nonfatal cancers. This conclusion is limited by the relatively short follow-up of the studies analyzed. Similar analyses of data from studies with longer follow-up periods would be valuable. PMID- 11403757 TI - Guar gum for body weight reduction: meta-analysis of randomized trials. AB - PURPOSE: To determine the efficacy of the dietary fiber guar gum as a therapeutic option for reducing body weight by conducting a meta-analysis of randomized controlled trials. METHODS: Literature searches were performed on the electronic databases Medline, Embase, Biosis, Amed, and the Cochrane Library. Manufacturers of commercial guar gum preparations and experts on the subject were contacted to provide any published or unpublished trials. For inclusion, trials had to state that they were randomized, double blinded, and placebo controlled, used guar gum monopreparations, and reported body weight as an endpoint. No language restrictions were imposed. Two reviewers independently extracted data in a standardized manner according to predefined criteria and evaluated methodological quality using the scoring system developed by Jadad. Discrepancies were settled through discussion. RESULTS: Thirty-four trials were identified and 20 could be included. Eleven trials provided data that were suitable for statistical pooling. The meta-analysis indicated a nonsignificant difference in patients receiving guar gum compared with patients receiving placebo (weighted mean difference -0.04 kg; 95% confidence interval (CI): -2.2 to 2.1). Analysis of six trials with similar methodologic features corroborates these findings (weighted mean difference -0.3 kg; 95% CI: -4.0 to 3.5). Adverse events most frequently reported were abdominal pain, flatulence, diarrhea, and cramps. Overall, 11 patients (3%) dropped out owing to adverse events. CONCLUSIONS: This meta-analysis suggests that guar gum is not efficacious for reducing body weight. Considering the adverse events associated with its use, the risks of taking guar gum outweigh its benefits for this indication. Therefore, guar gum cannot be recommended as a treatment for lowering body weight. PMID- 11403758 TI - Diagnostic dilemma. Vocal cord dysfunction. PMID- 11403759 TI - Diuresis and transudative effusions--changing the rules of the game. PMID- 11403760 TI - Hypercalcemia and sarcoidosis--another piece of the puzzle falls into place. PMID- 11403761 TI - Statins and cancer: a case of meta-uncertainty. PMID- 11403763 TI - Supply, demand, and the workforce of internal medicine. PMID- 11403764 TI - Dogs shed Neospora caninum oocysts after ingestion of naturally infected bovine placenta but not after ingestion of colostrum spiked with Neospora caninum tachyzoites. AB - An experiment was carried out to determine whether bovine colostrum or placenta could be a source of infection of Neospora caninum for dogs. For this purpose, two dogs were fed bovine colostrum to which culture-derived N. caninum tachyzoites were added and two other dogs were fed placental cotyledonary tissue from N. caninum seropositive cows. One dog served as a negative control during the start of the experiment but this control dog was fed cotyledonary tissue later on. None of the dogs did produce serum antibodies to N. caninum. All three dogs that were fed cotyledonary tissue did shed N. caninum oocysts, but no oocyst shedding was seen in the two dogs that were fed colostrum with N. caninum tachyzoites. Oocyst excretion did not resume in two dogs after repeated feeding of N. caninum infected placenta. The identity of the oocysts was confirmed by a bioassay in gerbils. It is concluded that ingestion of bovine placenta by dogs is an effective mode of transmission of N. caninum from cattle to dogs. PMID- 11403765 TI - DNA-based vaccines against malaria: status and promise of the Multi-Stage Malaria DNA Vaccine Operation. AB - The introduction of DNA vaccine technology has facilitated an unprecedented multi antigen approach to developing an effective vaccine against complex pathogens such as the Plasmodium spp. parasites that cause malaria. We have established the capacity of DNA vaccines encoding Plasmodium antigens to induce CD8(+) cytotoxic T lymphocyte and interferon-gamma responses in mice, monkeys and humans. However, like others, we have found that the first or second generation DNA vaccines on their own are not optimal, and have demonstrated the potential of heterologous prime/boost immunisation strategies involving priming with DNA and boosting with poxvirus or recombinant protein in adjuvant. In this review, we summarise the current status and promise of our programmatic efforts to develop a DNA-based vaccine against malaria, our Multi-Stage Malaria DNA Vaccine Operation, and illustrate the transition of promising developments in the laboratory to clinical assessment in humans. PMID- 11403766 TI - Tumour necrosis factor alpha receptors: role in the physiopathology of protozoan parasite infections. AB - Tumour necrosis factor alpha (TNF alpha) is an important cytokine in immune regulation and resistance to various micro-organisms. It provides signals to the target cells through two different receptors: TNFR1 and TNFR2. The present report reviews the role of TNF receptors (TNFRs) in the immune response against protozoan parasite infections of medical interest (Toxoplasma gondii, Leishmania major, Trypanosoma cruzi, Plasmodium spp.). TNF alpha has been regarded as a modulator cytokine in host defence against protozoans infections and recent findings on experimental gene-deficient mice have showed that TNF alpha/TNFRs pathway may be beneficial for host protection during these infections. PMID- 11403767 TI - Characterisation of an ATP diphosphohydrolase (Apyrase, EC 3.6.1.5) activity in Trichomonas vaginalis. AB - In the present report the enzymatic properties of an ATP diphosphohydrolase (apyrase, EC 3.6.1.5) in Trichomonas vaginalis were determined. The enzyme hydrolyses purine and pyrimidine nucleoside 5'-di- and 5'-triphosphates in an optimum pH range of 6.0--8.0. It is Ca(2+)-dependent and is insensitive to classical ATPase inhibitors, such as ouabain (1 mM), N-ethylmaleimide (0.1 mM), orthovanadate (0.1 mM) and sodium azide (5 mM). A significant inhibition of ADP hydrolysis (37%) was observed in the presence of 20 mM sodium azide, an inhibitor of ATP diphosphohydrolase. Levamisole, a specific inhibitor of alkaline phosphatase, and P(1), P(5)-di (adenosine 5'-) pentaphosphate, a specific inhibitor of adenylate kinase, did not inhibit the enzyme activity. The enzyme has apparent K(m) (Michaelis Constant) values of 49.2+/-2.8 and 49.9+/-10.4 microM and V(max) (maximum velocity) values of 49.4+/-7.1 and 48.3+/-6.9 nmol of inorganic phosphate x min(-1) x mg of protein(-1) for ATP and ADP, respectively. The parallel behaviour of ATPase and ADPase activities and the competition plot suggest that ATP and ADP hydrolysis occur at the same active site. The presence of an ATP diphosphohydrolase activity in T. vaginalis may be important for the modulation of nucleotide concentration in the extracellular space, protecting the parasite from the cytolytic effects of the nucleotides, mainly ATP. PMID- 11403768 TI - Cytochemical localisation of calcium ATPase activity during the erythrocytic cell cycle of Plasmodium falciparum. AB - Using a cytochemical technique, we evaluated the levels of Ca(2+)-ATPase activity in the plasmatic and in the parasitophorous vacuole membrane through the different developmental stages of the Plasmodium falciparum parasitised erythrocyte. We found that the activity is detectable and remains unaltered in the plasma membrane throughout the 48 h cell cycle. However, in the parasitophorous membrane, although the activity was very similar to that measured in the plasma membrane of the young stages (younger than 20-h-old parasites), it diminished gradually with maturation and in schizonts it was almost undetectable. These data suggest that the plasma membrane Ca(2+)-ATPase is important in the maintenance of a low erythrocyte cytoplasmic Ca(2+) concentration, and that in addition it could be a way to supply the vital cation to the parasite at the beginning of the infection, when other transport mechanisms have not yet developed. PMID- 11403769 TI - Immunomicroscopical observations on the nervous system of adult Eudiplozoon nipponicum (Monogenea: Diplozoidae). AB - Neuronal pathways have been examined in adult Eudiplozoon nipponicum (Monogenea: Diplozoidae), using cytochemistry interfaced with confocal scanning laser microscopy, in an attempt to ascertain the status of the nervous system. Peptidergic and serotoninergic innervation was demonstrated by indirect immunocytochemistry and cholinergic components by enzyme cytochemical methodology; post-embedding electron microscopical immunogold labelling revealed neuropeptide immunoreactivity at the subcellular level. All three classes of neuronal mediators were identified throughout both central and peripheral elements of a well-differentiated orthogonal nervous system. There was considerable overlap in the staining patterns for cholinergic and peptidergic components, while dual immunostaining revealed serotonin immunoreactivity to be largely confined to a separate set of neurons. The subcellular distribution of immunoreactivity to the flatworm neuropeptide, GYIRFamide, confirmed neuropeptide localisation in dense-cored vesicles in the majority of the axons and terminal varicosities of both central and peripheral nervous systems. Results reveal an extensive and chemically diverse nervous system and suggest that pairing of individuals involves fusion of central nerve elements; it is likely also that there is continuity between the peripheral nervous systems of the two partner worms. PMID- 11403770 TI - The susceptibility of grayling (Thymallus thymallus) to experimental infections with the monogenean Gyrodactylus salaris. AB - The pathogenic monogenean Gyrodactylus salaris infecting Atlantic salmon (Salmo salar) is found to attach and reproduce under laboratory conditions on several species in the subfamily Salmoninae other than the Atlantic salmon. The gyrodactylid species Gyrodactylus thymalli infecting grayling (Thymallus thymallus) in another subfamily, Thymallinae, is previously said to be very similar to G. salaris based on morphometry and genetical analysis which prompted the present laboratory experiments to test the susceptibility and resistance of grayling to G. salaris. All 0+ and 1+ grayling became infected with G. salaris during the experimental infection procedure. However, both innate resistant and susceptible grayling were found. In susceptible individually isolated fish, parasite reproduction lasted for more than 35 days. Parasite reproduction also occurred among grouped grayling as judged from the duration of infection of more than 50 days. However, grayling susceptibility as judged from G. salaris reproduction, was very limited. Hence, the results indicate significant biological differences between the function of Atlantic salmon and grayling as host for G. salaris. The grayling is interpreted as unable to sustain G. salaris in nature which implies that G. thymalli is not conspecific with G. salaris. However, G. salaris dispersal by grayling cannot be excluded. PMID- 11403771 TI - Role of oligosaccharides in the immune response of sheep vaccinated with Lucilia cuprina larval glycoprotein, peritrophin-95. AB - The larvae of the fly Lucilia cuprina cause a cutaneous myiasis in mammalian hosts, particularly sheep. The glycoprotein, peritrophin-95, isolated from Lucilia cuprina larval peritrophic matrix, is a candidate vaccine antigen. This protein induced an immune response in vaccinated sheep that inhibited larval growth. Recombinant forms of peritrophin-95 were produced in bacteria and baculovirus-infected insect cells. The bacterial protein was not glycosylated and incorrectly folded whereas the insect cell-expressed protein was glycosylated and probably correctly folded. Sheep immunised with purified native peritrophin-95 generated strong larval growth inhibitory activity in their sera, whereas sheep immunised with either recombinant form of peritrophin-95 generated only relatively weak inhibitory activity. Ingested ovine antibodies to native peritrophin-95 mediated the anti-larval growth activity and this was independent of the presence of ovine complement. The activity was associated with IgG(1) and IgG(2) but not IgM. There were strong antibody responses to both the correctly folded native peritrophin-95 polypeptide and the oligosaccharides present on this glycoprotein. Immuno-affinity isolation of antibody to the peritrophin-95 polypeptide and antibody to peritrophin-95 oligosaccharides demonstrated that the larval growth inhibitory activity resided with both antibodies. Lectin blots and ELISA data showed substantial differences between the oligosaccharides attached to native peritrophin-95 and insect cell-expressed recombinant peritrophin-95. It was concluded that the oligosaccharides attached to native peritrophin-95 and its unique polypeptide structure are essential for the induction of larval growth inhibitory activity in the sera of sheep vaccinated with this antigen. PMID- 11403772 TI - Effect of sand fly saliva on Leishmania uptake by murine macrophages. AB - Leishmania promastigotes are introduced into the skin by blood-sucking phlebotomine sand flies. In the vertebrate host, promastigotes invade macrophages, transform into amastigotes and multiply intracellularly. Sand fly saliva was shown to enhance the development of cutaneous leishmaniasis lesions by inhibiting some immune functions of the host macrophages. This study demonstrates that sand fly saliva promotes parasite survival and proliferation. First, macrophages gravitated towards increasing concentrations of sand fly saliva in vitro. Secondly, saliva increased the percentage of macrophages that became infected with Leishmania promastigotes and exacerbated the parasite load in these cells. Thus, during natural transmission, saliva probably reduces the exposure of promastigotes to the immune system by attracting macrophages to the parasite inoculation site and by accelerating the entry of promastigotes into macrophages. Saliva may also enhance lesion development by shortening the generation time of dividing intracellular amastigotes. PMID- 11403773 TI - Differentiation of two locally sympatric Protopolystoma (Monogenea: Polystomatidae) species by temperature-dependent larval development and survival. AB - The developmental response of egg stages to different environmental temperature regimes was studied in Protopolystoma xenopodis and Protopolystoma orientalis (Monogenea: Polystomatidae) isolates from southern Africa. Eggs failed to develop at 10 degrees C, whilst at 15 degrees C only P. xenopodis completed larval development, hatching 49--88 days post-collection. Respective hatching windows were 26--34 (P. xenopodis) and 37--49 (P. orientalis) days at 20 degrees C, and 18--26 and 27--37 days at 25 degrees C. Continuous maintenance at 30 degrees C was lethal for eggs of both species. There were no consistent interspecific differences in the response of egg stages to low and high temperature shocks during early embryonic development. PMID- 11403774 TI - Human giardiasis: genotype linked differences in clinical symptomatology. AB - Giardia duodenalis infection in humans can cause a variety of clinical symptoms. The relation between clinical symptomatology and the Giardia isolate genotype was studied in 18 Dutch patients infected with G. duodenalis who visited their general practitioner. Contrary to earlier studies, a 100% correlation between severity of diarrhoeal complaints and genotype was found: assemblage A isolates were solely detected in patients with intermittent diarrhoeal complaints, while assemblage B isolates were present in patients with persistent diarrhoeal complaints. These results are significant because they show for the first time that genetically linked features of G. duodenalis are major determinants in the severity of infection in human giardiasis. PMID- 11403775 TI - Molecular characterisation of a predominant antigenic region of Giardia lamblia variant surface protein H7. AB - During infection, the intestinal protozoan parasite Giardia lamblia undergoes continuous antigenic variation which is determined by diversification of the parasite's major surface antigen, named VSP (variant surface protein). One member from this protein family, VSP H7, is expressed by G. lamblia clone GS/M-83-H7. In the present study, we characterised a highly antigenic portion of VSP H7 which is positioned inside a 130 amino acid C-terminal region of the protein. This region overlaps with a cysteine-rich motif that is rather conserved within the VSP family. Detailed molecular dissection of the antigenic portion monitored a 12 amino acid peptidyl structure which constitutes a non-conformational epitope of VSP H7. In the murine host, this epitope is recognised relatively early (before day 10 p.i.) during infection and stimulates a strong intestinal immunoglobulin A response. At late infective stages (after day 10 p.i.) this immune reaction is progressively complemented by reactions against 'late' antigenic epitopes which are also located inside the 130 amino acid antigenic portion but in closer proximity to the C-terminal end of VSP H7 than the 12 amino acid epitope. Both the high antigenicity and the conserved character suggest that the 12 amino acid epitope is a key factor within the immunological interplay between G. lamblia and the experimental murine host. PMID- 11403776 TI - A molecular phylogeny of the genus Ichthyocotylurus (Digenea, Strigeidae). AB - Three nucleotide data sets, two nuclear (ribosomal internal transcribed spacer regions 1 and 2, ITS1 and ITS2) and one mitochondrial (cytochrome c oxidase subunit 1, CO1), were analysed using distance matrix and maximum likelihood methods to determine the inter-relationships amongst the four species attributed to the genus Ichthyocotylurus Odening, 1969. Sequence data obtained from all gene loci investigated supported the position of Ichthyocotylurus variegatus as a species discrete from Ichthyocotylurus platycephalus. Phylogenetic analyses yielded congruent trees, with I. variegatus isolates comprising a common clade to which I. platycephalus constitutes a sister taxon. Ichthyocotylurus erraticus and Ichthyocotylurus pileatus were found to demonstrate a similarly close inter specific relationship. The greatest intra-generic divergence occurred in the CO1 region (16% variability), with resultant disparities in three to eight encoded amino acids. PCR amplification yielded multiple ITS1 products for all Ichthyocotylurus spp. Analyses of equivalent-sized amplicons showed 5.4% intra generic variation and several point mutations between I. variegatus isolates from different geographical localities and from different piscine hosts. The ITS2 locus was extremely conserved, with less than 1% variation between species. No intra-specific variation was recorded for any CO1 or ITS2 sequences. PMID- 11403777 TI - The striped skunk (Mephitis mephitis) is an intermediate host for Sarcocystis neurona. AB - Striped skunks, initially negative for antibodies to Sarcocystis neurona, formed sarcocysts in skeletal muscles after inoculation with S. neurona sporocysts collected from a naturally infected Virginia opossum (Didelphis virginiana). Skunks developed antibodies to S. neurona by immunoblot and muscles containing sarcocysts were fed to laboratory-reared opossums which then shed sporulated Sarcocystis sporocysts in their faeces. Mean dimensions for sporocysts were 11.0 x 7.5 microm and each contained four sporozoites and a residuum. Sarcocysts from skunks and sporocysts from opossums fed infected skunk muscle were identified as S. neurona using PCR and DNA sequence analysis. A 2-month-old, S. neurona-naive pony foal was orally inoculated with 5 x 10(5) sporocysts. Commercial immunoblot for antibodies to S. neurona performed using CSF collected from the inoculated pony was low positive at 4 weeks p.i., positive at 6 weeks p.i., and strong positive at 8 weeks p.i. Gamma-interferon gene knockout mice inoculated with skunk/opossum derived sporocysts developed serum antibodies to S. neurona and clinical neurologic disease. Merozoites of S. neurona present in the lung, cerebrum, and cerebellum of mice were detected by immunohistochemistry using polyclonal antibodies to S. neurona. Based on the results of this study, the striped skunk is an intermediate host of S. neurona. PMID- 11403780 TI - Lack of positive interaction between CO2 and hypoxic stimulation for P(CO2)-VAS response slope in humans. AB - To compare the effect of hypoxia on ventilatory responses and respiratory sensation to carbon dioxide, 29 young adults were examined using a modified Read's rebreathing method with four experimental conditions. We used varying gas mixtures and kept PET(O2) constant at >300, 100, 80 and 60 mmHg for each four rebreathing tests. Respiratory sensation was measured by visual analog scale (VAS). The slope of the CO2-ventilation response curve increased significantly with hypoxia, confirming a positive ventilatory interaction between hypoxia and hypercapnia. However, the slope of the CO2-VAS response curve remained unchanged. The V(E)-VAS relation slope tended to become depressed with advancing hypoxia, i.e. the magnitude of VAS elicited by a given ventilation decreased with hypoxia, signifying that dyspneic sensation was effectively mitigated during hypoxic hyperventilation. We suggest that this relief of dyspneic sensation might be due to the inhibitory respiratory effect from augmented pulmonary stretch receptor (PSR) activity. PMID- 11403781 TI - Effects of embryonic CO2 exposure on the adult ventilatory response in quail: does gender matter? AB - To test the hypothesis that the adult ventilatory response to CO2 can be modified by exposure to CO2 during development, we exposed Japanese quail (Coturnix japonica) from two populations to 2% CO2 throughout embryonic development. The ventilatory response to 6% CO2 was subsequently measured in control and CO2 exposed quail following a 6-9 week deacclimation period. In one population of quail, CO2-exposed female, but not male, quail had a reduced ventilatory response to 6% CO2 as adults (P<0.001). Although control and CO2-exposed quail had similar ventilation while breathing air, CO2-exposed females exhibited a 26% reduction in mean ventilation at 6% CO2, primarily due to a reduced tidal volume. In contrast, the identical incubation treatment had no effect on the ventilation of either gender in a second population of quail. It appears that developmental plasticity in the hypercapnic ventilatory response may generally be greater in females, although this conclusion may vary depending on genetic factors. PMID- 11403782 TI - NO and the hypometabolic and hypothermic responses to hypoxia in the rat. AB - Nitric oxide (NO) has been identified as a possible mediator of the thermoregulatory and respiratory responses to hypoxia. The present study was designed to assess the effects of an NO donor (S-nitroso-N-acetylpenicillamine, SNAP) injected systemically (0.5-2.0 mg/kg) in unanesthetized adult rats studied at ambient temperatures (Ta) of 26, 24, and 15 degrees C. The metabolic rate (VO2), ventilation (V), and colonic temperature (Tc) were recorded while the animals were exposed to normoxia or to hypoxia (FI(O2)=0.11). During normoxia, at all values of Ta, SNAP increased VO2 and V and the decrease in Tc observed following saline injection was prevented or attenuated. However, SNAP did not modify the hypometabolic, hypothermic, and hyperventilatory responses to hypoxia. We concluded that, confirming previous studies, NO may play a role in the control of VO2 and V in normoxia. The failure to observe any effect following the systemic NO donor injection during hypoxia may indicate opposing actions of the drug at different sites resulting in no net changes in ventilatory and metabolic responses to hypoxia. PMID- 11403783 TI - Factors influencing the shape of the inspiratory flow. AB - An index (IS), quantitating the departure of the inspiratory flow profile (IFP) from the rectangular one, i.e. the optimal profile (IS=0), was computed from pneumotachograms recorded in 26 normal, anesthetized subjects breathing spontaneously through the endotracheal tube (ETT) or face mask (FM) with or without added resistances (R) and in 27 awake subjects breathing through the mouth and FM in the supine and seated posture at rest and during exercise (40 W) on a cycloergometer, through the nose and FM, and through the mouthpiece (MP). During anesthesia, IS decreased with R both while breathing through the ETT (DeltaIS=-0.037+/-0.006 (SE); P<0.001) and FM (DeltaIS=-0.054+/-0.008; P<0.001). This indicates that (a) the change of IFP towards the optimal shape is reflex in nature and related to the dynamic inspiratory load, and (b) tracheobronchial mechanoreceptors and inspiratory muscles are involved in this response. The reflex is also operative in awake subjects, since IS decreased whenever the inspiratory dynamic load was increased, as on turning from seated to supine posture (DeltaIS=-0.024+/-0.003; P<0.001), shifting from mouth to nose breathing (DeltaIS=-0.034+/-0.003; P<0.05), from rest to mild exercise (DeltaIS=-0.066+/ 0.005; P<0.001). The different IS value between FM and MP breathing (DeltaIS=0.036+/-0.004; P<0.001) indicates, however, that other factors, likely behavioral, also affect the IFP. PMID- 11403784 TI - Convection requirement is established by total metabolic rate in the newborn tammar wallaby. AB - Ventilation (VE) and metabolic rate, determined from both pulmonary and cutaneous gas exchange, were measured in 39 newborn tammar wallabies, Macropus eugenii, aged between 0 and 3 days. In 1-day-old animals both total metabolic rate (skin+lung exchange) and ventilation were approximately 50% of the values predicted for eutherian newborns of equivalent body mass. Hence, the convection requirement (VE/total metabolic rate) of the newborn tammar is close to predicted values for newborns and adult mammals in general. Metabolic rate in the newborn tammar is supported to some extent by cutaneous gas exchange, approximately 30% of the total in the 1-day-old animal. This ratio diminishes with increasing age as the lung takes on an increasingly more important role for respiratory exchange. The early establishment of the convection requirement in the newborn tammar, despite significant cutaneous gas exchange, provides supporting evidence that metabolic rate per se is important in establishing the level of ventilation. PMID- 11403785 TI - Circadian patterns in men acclimatized to intermittent hypoxia. AB - Six men, normally working shifts of 7 days at high altitude (HA, 3800 m, approximately 480 mm Hg barometric pressure) followed by 7 days of rest at sea level (SL), were studied during the last days of their HA and SL shifts with a 24 h constant routine protocol of sustained wakefulness and minimal activity. The amplitude of the circadian oscillations of oxygen consumption, breathing rate, thoracic skin temperature and diastolic pressure did not differ between HA and SL. At HA, the amplitude of the tympanic and calf temperature oscillations, were, respectively, lower and higher than at SL. End-tidal P(CO2) and systolic pressure had larger amplitude oscillations at HA than at SL. Hence, also in humans, as previously shown in animals, hypoxia can affect some circadian patterns, including those involved in thermoregulation. These effects of hypoxia could contribute to sleep disturbances at HA and in patients with cardiorespiratory diseases. PMID- 11403786 TI - Oscillation of the lung by chest-wall vibration. AB - Vibration of the thoracic surface has been shown to modify the drive to breathe and the sensation of dyspnea. It has been suggested that respiratory muscle afferents generate these effects. The possibility that the consequences of chest wall vibration also involve intra-pulmonary afferents led us to investigate whether such vibration reaches the airways. Two vibratory stimuli were independently applied to four chest-wall sites and two control sites on eight healthy subjects. During separate breath holds, the vibrator was held on each site while subjects periodically opened and closed the pharynx. Airway pressure (P(AW)) was measured at the mouth. Spectral analysis of P(AW) showed pressure oscillations occurred at the same frequency as that of the vibrators when the pharynx was open; oscillation amplitude was vastly reduced when the pharynx was closed. Oscillation amplitude was also significantly larger during vibration at greater amplitude. These data demonstrate that vibration over the chest-wall vibrates the lung and could potentially excite intrapulmonary receptors. PMID- 11403787 TI - 'Respiratory' oscillations of cardiovascular parameters during voluntary apnea. AB - The aim of this study was to ascertain the persistence of heart rate and blood pressure oscillations at the onset of voluntary apnea in humans. In 24 young subjects (10 males, 14 females, mean age 20.4 years) RR intervals, systolic blood pressure (SBP) and diastolic blood pressure (DBP) during controlled breathing (CB) of atmospheric air followed by voluntary apnea on FRC level were recorded continuously. The cosine functions were then fitted by nonlinear regression analysis to heart rate, SBP and DBP oscillations during CB and at the onset of apnea. During apnea, the changes of base of RR intervals indicate primary tachycardic reaction followed by a decrease in heart rate. The base of the blood pressure oscillations was higher (hypertensive reaction) in apnea. At least one cosine-like wave oscillation was present at the onset of apnea in heart rate, SBP and DBP and the second wave was present in all assessed parameters at least in 70% of recordings. There were no significant gender differences neither in the duration of breath holding nor in the RR and SBP oscillations parameters. PMID- 11403788 TI - The optic nerve head component of the monkey's (Macaca mulatta) multifocal electroretinogram (mERG). AB - To search for an optic nerve head component (ONHC) in the monkey's (Macaca mulatta) multifocal electroretinogram (mERG), mERGs from three animals were recorded with different electrode configurations. A component with a latency that varied with distance from the optic nerve head was easily identified by eye in recordings from the speculum of a Burian-Allen electrode referenced to a DTL on the unstimulated eye. This component was reasonably well isolated by subtracting a weighted version of a Burian-Allen bipolar recording or by employing the extraction algorithm of Sutter and Bearse (1999, Vision Research, 39, 419-436). The waveform of this component resembles the ONHC reported for the human mERG. PMID- 11403789 TI - Gecko vision--retinal organization, foveae and implications for binocular vision. AB - Geckos comprise both nocturnal and diurnal genera, and between these categories there are several transitions. As their retinae have definitely to be classified as pure cone retinae, they provide an especially attractive model for comparison of organization and regional specializations adapted to very different photic environments. While the visual cells themselves show clear adaptations to nocturnal or diurnal lifestyles, the overall retinal organization is more related to that of diurnal vertebrates. Nocturnal geckos have lost any foveae of their diurnal ancestors, but they have retained a low convergence ratio and a high visual cell density. To enhance visual sensitivity, they exploit binocular - but not necessarily stereoscopic - vision. Diurnal species have retained binocular vision. Most diurnal species have developed new foveae, which are consequently located not in the central but in the temporal region of the retina. PMID- 11403790 TI - Neural synergy in visual grouping: when good continuation meets common fate. AB - A modified version of the 'path finder' display consisting of many small oriented Gabor patches was used to study the joint contributions of spatial and temporal structures to shape perception. A two-interval forced-choice procedure measured detectability of curved 'paths' defined by orientation ('good continuation') and/or by temporal synchrony of change in motion direction ('common fate'). When orientation was completely random (no spatial 'path' cue) temporal synchrony still supported reliable performance, but only when correlation of change among 'path' elements was high. When combined, these two weak spatial and temporal structures yielded performance in excess of probability summation: 'paths' weakly defined by orientation were highly conspicuous when the constituent Gabors underwent synchronized changes in direction of motion, even though the individual directions of path elements were uncorrelated. Spatial grouping from temporal structure may arise from correlated transients associated with synchronized changes in motion direction. Evidently these two mechanisms for promotion of spatial grouping interact synergistically. PMID- 11403791 TI - Target/surround asymmetry in perceptual filling-in. AB - Four experiments examined how differences in the properties of the target and surround affect the time required for perceptual filling-in. They examined differences in luminance, orientation, spatial frequency, and color. A larger target/surround difference delayed filling-in ('feature difference effect'). Interestingly, exchanging the target and surround properties significantly varied the time ('target/surround asymmetry'). Filling-in was facilitated when the target was brighter and closer to the vertical or horizontal than the surround. Little asymmetry was found in the frequency domain, while significant asymmetry was observed for specific color combinations. These effects are discussed with respect to edge adaptation, feature adaptation, balance of neural activities, and contextual modulation. PMID- 11403792 TI - Pulse and steady-pedestal contrast discrimination: effect of spatial parameters. AB - The goal of this study was to establish the spatial summation properties associated with inferred PC- and MC-pathway mediated psychophysical contrast discrimination. Previous work has established two paradigms that reveal characteristic signatures of these pathways. In the pulse paradigm, a four-square array was pulsed briefly, on a constant background. In the steady-pedestal paradigm, the stimulus array was presented continuously as a steady-pedestal within a constant surround. In both paradigms, one square differed from the others, giving the observer a forced choice spatial discrimination task. Area summation functions derived for the pulse paradigm decreased with area, with a slope of -0.25 on a log-log axis. Area summation functions derived for the steady pedestal paradigm decreased as a power function of area, approaching an asymptote above one square degree. The latter are consistent with the classical data of threshold spatial summation. PMID- 11403793 TI - Temporal resolution of orientation-based texture segregation. AB - We analysed the temporal-frequency characteristics of two functional processes involved in orientation-based texture segregation: local orientation coding and subsequent orientation-contrast coding. Two texture images, in which each micropattern was rotated by 90 degrees, were alternated at various temporal frequencies. A micropattern was a second-derivative (D2) of a Gaussian that loses orientation information when temporally fused with the orthogonal D2 pattern. We measured the upper temporal-frequency limits for localising the target region whose mean orientation differed from the background by 90 degrees or by 45 degrees. If the temporal limit of the texture perception is determined by the most sluggish processing stage, the temporal limit for the 90 degrees texture should be determined by local orientation coding or by orientation-contrast coding, depending on which stage has the lower temporal precision. On the other hand, the 45 degrees texture should always be segregated below the temporal limit of local orientation coding regardless of the temporal limit of orientation contrast coding. We found that the temporal limit for the 90 degrees texture was slightly higher than that for the 45 degrees texture under spatial conditions appropriate for texture segregation. Moreover, an orientation-noise analysis of segregation performance for a wide range of temporal frequencies revealed that the temporal-frequency sensitivities for the two textures were nearly identical. These results imply that the temporal limit for orientation-based texture segregation depends only on that of local orientation coding. This conclusion further suggests that the potential temporal resolution of orientation-contrast coding is not lower than that of local orientation coding, which would imply that the orientation-contrast coding is unlikely to be mediated by sluggish neural processes. PMID- 11403794 TI - Filling-in the details on perceptual fading. AB - We examined the perceptual disappearance (or 'filling in') of a peripheral target surrounded by dynamic texture. Targets defined by different visual attributes were used to explore the importance of target properties in determining the time course of fading. Introducing luminance-, motion- or direction-contrast between the target and background increased the time-to-fade. For motion contrast, this was related to target visibility. Targets defined by a difference of texture from the background took longer to fade than those defined by a difference of motion. This might correspond to activity in different visual areas, or could be due to different visibilities in each case. PMID- 11403795 TI - Perceived length in the central visual field: evidence for visual field asymmetries. AB - Visual performance for judging the length of a simultaneously presented pair of radial lines, reciprocally opposed by 180 degrees at a central fixation point, was assessed for 24 radial positions of test lines, for three viewing conditions (binocular, left and right monocular) and five different standard line sizes (1.43-7.13 degrees ). Generally, the results showed underestimation of the test line. Furthermore, clear visual field asymmetries were observed between the upper versus lower visual fields and the left versus right visual fields with greater underestimation for test lines presented in the lower and right visual fields. Also, asymmetries tended to be strongest along the 30 and 150 degrees radial orientations. Fourier analysis indicated that these asymmetries are mainly described by summing up the f0, f1, f2 and f5 components. PMID- 11403796 TI - Three-dimensional eye position during static roll and pitch in humans. AB - We investigated how three-dimensional (3D) eye position is influenced by static head position relative to gravity, a reflex probably mediated by the otolith organs. In monkeys, the torsional component of eye position is modulated by gravity, but little data is available in humans. Subjects were held in different head/body tilts in roll and pitch for 35 s while we measured 3D eye position with scleral coils, and we used methods that reduced torsion artifacts produced by the eyelids pressing on the contact lens and exit wire. 3D eye positions were described by planar fits to the data (Listing's plane), and changes in these planes showed how torsion varied with head position. Similar to findings in monkeys, the eyes counterrolled during roll tilts independent of horizontal and vertical eye position, reaching a maximum torsion of 4.9 degrees. Counterroll was not proportional to the shear force on the macula of the utricles: gain (torsion/sine of the head roll angle) decreased by 50% from near upright to ear down. During pitch forward, torsion increased when subjects looked right, and decreased when they looked left. However, the maximum change of torsion was only 0.06 degrees per degree of horizontal eye position, which is less than reported in monkey. Also in contrast to monkey, we found little change in torsion when subjects were pitched backwards. PMID- 11403797 TI - Isolating the M(y)-cell response in dyslexia using the spatial frequency doubling illusion. AB - The contribution of M(y)-cell activity within a framework of a magnocellular deficit theory of dyslexia is currently unknown. Twenty-one dyslexic readers and 19 control readers were compared on their threshold detection for the frequency doubling illusion - an index of M(y)-cell activity, coherent motion, and a visual acuity task. The dyslexic group performed more poorly on detection of the frequency doubling illusion and coherent motion compared to the control group, but both groups performed comparably on the visual acuity task. The results from this study indicate that if a magno deficit exists in dyslexia, it may originate at a retinal level at least partly mediated by M(y)-cell abnormalities. PMID- 11403798 TI - The premotor region essential for rapid vertical eye movements shows early involvement in Alzheimer's disease-related cytoskeletal pathology. AB - The rostral interstitial nucleus of the medial longitudinal fascicle (riMLF) contains premotor neurons essential for the generation of rapid vertical eye movements. The Alzheimer's disease (AD)-related cytoskeletal changes and beta amyloid deposits in this nucleus were examined in 30 autopsy cases and compared to the involvement of three associated nuclei - Edinger-Westphal nucleus, nucleus of Darkschewitsch and interstitial nucleus of Cajal. The riMLF displays slight cytoskeletal alterations already in the early stages in the development of the cortical cytoskeletal pathology (cortical NFT/NT-stages I-II, representing the preclinical phase of AD). In the cortical NFT/NT-stages III-IV (i.e. incipient phase of AD), the cytoskeletal pathology in the riMLF is pronounced and in stages V-VI (i.e. clinical phase of AD) it is severe. The progression of the cytoskeletal pathology in the riMLF correlates significantly with the cortical NFT/NT-stages I-VI that reflect the clinical course of AD. Isolated beta-amyloid deposits appear in the riMLF for the first time in the final beta-amyloid stage. In the Edinger-Westphal nucleus, in the nucleus of Darkschewitsch and most markedly in the interstitial nucleus of Cajal, the pathological changes were significantly less severe than those in the riMLF. In the event that the cytoskeletal pathology impairs the function of the premotor neurons of the riMLF, one would predict a progressive slowing of vertical saccades corresponding to the advancing cortical NFT/NT-stages. PMID- 11403799 TI - Reaction time in automated kinetic perimetry: effects of stimulus luminance, eccentricity, and movement direction. AB - PURPOSE: To determine the effects of stimulus eccentricity and luminance level on the reaction time (RT) of young normal volunteers during automated kinetic campimetry. METHODS: We used a specially designed video-campimetric device equipped with a continuous infrared (IR) pupillographic fixation control (Tubingen Computer Campimeter) and recorded reaction times upon presenting horizontally moving small circular stimuli (size 26'; constant angular velocity 2 degrees /s) starting at 16 locations within the central 30 degrees -radius of the visual field. Two different levels of stimulus luminance were used (41.6 cd/m(2) and 110 cd/m(2)), while background luminance was 10 cd/m(2). Each stimulus was presented a total of six times in a randomized order. Subjects were 12 healthy young individuals (aged 21-30 years) with normal ophthalmic examinations. An analysis of variance (ANOVA) was performed on the data. RESULTS: RTs showed considerable inter- and intra-individual variation with individual least squares means (LSM, fitted values of a linear model) ranging from 305 to 454 ms, and residual standard deviation (R.S.D.) 66 ms. Reaction times did not differ significantly as a function of stimulus direction (P>0.6). Higher luminance levels produced significantly reduced reaction times for all stimulus locations and directions (mean reduction: 16 ms; P<0.0001). Reaction times increased with increasing eccentricity, in the mean by 1.8 ms per degree of visual angle, from 365+/-4 ms (S.E.M.) foveally, to 407+/-2 ms at 30 degrees eccentricity; (P<0.0001). CONCLUSIONS: Automated kinetic perimetry should be designed to cope with significant, variable interindividual response characteristics. Other stimulus related factors, such as eccentricity or luminance level, have a significant but comparatively small effect on reaction time within the central 30 degrees -radius visual field in healthy young individuals. PMID- 11403800 TI - How consumers can and should improve clinical trials. PMID- 11403801 TI - Endarterectomy or angioplasty for treatment of carotid stenosis? PMID- 11403802 TI - Use of a specific aromatase inhibitor in delayed puberty. PMID- 11403803 TI - Suppression of unwanted memories: repression revisited? PMID- 11403804 TI - Chemokine receptor polymorphism in transplantation immunology: no longer just important in AIDS. PMID- 11403805 TI - Vagal-nerve stimulation for epilepsy. PMID- 11403806 TI - Bringing it all together: Lancet-Cochrane collaborate on systematic reviews. PMID- 11403807 TI - Immunology approaches the bedside. PMID- 11403808 TI - Endovascular versus surgical treatment in patients with carotid stenosis in the Carotid and Vertebral Artery Transluminal Angioplasty Study (CAVATAS): a randomised trial. AB - BACKGROUND: Percutaneous transluminal angioplasty and stenting (endovascular treatment) can be used to treat carotid stenosis, but risks and benefits are uncertain. We therefore compared endovascular treatment with conventional carotid surgery. METHODS: In a multicentre clinical trial, we randomly assigned 504 patients with carotid stenosis to endovascular treatment (n=251) or carotid endarterectomy (n=253). For endovascular patients treated successfully, we used stents in 55 (26%) and balloon angioplasty alone in 158 (74%). An independent neurologist followed up patients. Analysis was by intention to treat. FINDINGS: The rates of major outcome events within 30 days of first treatment did not differ significantly between endovascular treatment and surgery (6.4% vs 5.9%, respectively, for disabling stroke or death; 10.0% vs 9.9% for any stroke lasting more than 7 days, or death). Cranial neuropathy was reported in 22 (8.7%) surgery patients, but not after endovascular treatment (p<0.0001). Major groin or neck haematoma occurred less often after endovascular treatment than after surgery (three [1.2%] vs 17 [6.7%], p<0.0015). At 1 year after treatment, severe (70-99%) ipsilateral carotid stenosis was more usual after endovascular treatment (25 [14%] vs seven [4%], p<0.001). However, no substantial difference in the rate of ipsilateral stroke was noted with survival analysis up to 3 years after randomisation (adjusted hazard ratio=1.04, 95% CI 0.63-1.70, p=0.9). INTERPRETATION: Endovascular treatment had similar major risks and effectiveness at prevention of stroke during 3 years compared with carotid surgery, but with wide CIs. Endovascular treatment had the advantage of avoiding minor complications. PMID- 11403809 TI - Helicobacter pylori and symptomatic relapse of gastro-oesophageal reflux disease: a randomised controlled trial. AB - BACKGROUND: There is little information on the effects of Helicobacter pylori eradication in patients with a primary diagnosis of gastro-oesophageal reflux disease (GORD). Our aim was to investigate the effect of H pylori eradication in this group of patients. METHODS: We did a double-blind, randomised, placebo controlled study in 70 patients with GORD. We assigned individuals to three groups. All patients received lansoprazole 30 mg twice daily for 10 days, followed by 30 mg once daily for 8 weeks. Patients infected with H pylori received either antibiotics (clarithromycin 500 mg and amoxicillin 1000 mg twice daily) or placebo for the first 10 days. Controls were patients not infected with H pylori. Patients were followed up for 6 months at 2-week intervals for GORD symptoms. At the end of the study we repeated endoscopy and oesophageal and gastric 24 h-pH monitoring. FINDINGS: 58 of 70 patients completed our study. At the end of the study 16 of these patients were H pylori-positive (14 placebo and two eradication failures), 13 were negative because of successful H pylori eradication, and 29 were controls. H pylori-positive patients relapsed earlier (54 days) than did those in whom H pylori was eradicated (100 days) (p=0.046). The H pylori-negative control group relapsed after the longest period (110 days). However, time to relapse was also affected by oesophagitis grade (no oesophagitis 127 days, grade III or IV oesophagitis 18 days). When results were corrected for the affect of oesophagitis grade, H pylori-positive patients relapsed earlier (p=0.086) than H pylori-eradiated patients and controls (p=0.001). INTERPRETATION: H pylori infection positively affects the relapse rate of GORD. Eradication of H pylori could, therefore, help to prolong disease-free interval in patients with GORD. PMID- 11403810 TI - A specific aromatase inhibitor and potential increase in adult height in boys with delayed puberty: a randomised controlled trial. AB - BACKGROUND: The role of oestrogens in the closure of growth plates in both sexes is unequivocal. We postulated that inhibition of oestrogen synthesis in boys with delayed puberty would delay maturation of the growth plates and ultimately result in increased adult height. METHODS: We did a randomised, double-blind, placebo controlled study in which we treated boys with constitutional delay of puberty with testosterone and placebo, or testosterone and letrozole. Boys who decided to wait for the spontaneous progression of puberty without medical intervention composed the untreated group. FINDINGS: Letrozole effectively inhibited oestrogen synthesis and delayed bone maturation. Progression of bone maturation was slower in the letrozole group than in the placebo group. In 18 months, bone age had advanced 1.1 (SD 0.8) years in the untreated group and 1.7 (0.9) years in the group treated with testosterone and placebo, but only 0.9 (0.6) years in the letrozole group (p=0.03 between the treatment groups). Predicted adult height did not change significantly in the untreated group and in the placebo group, whereas in the group treated with letrozole the increase was 5.1 (3.7) cm (p=0.004). INTERPRETATIONS: Our findings suggest that if oestrogen action is inhibited in growing adolescents, adult height will increase. This finding provides a rationale for studies that aim to delay bone maturation in several growth disorders. PMID- 11403811 TI - Association between atopic dermatitis and insulin-dependent diabetes mellitus: a case-control study. AB - BACKGROUND: Up to two-thirds of children with atopic dermatitis have IgE-mediated allergic reactions and a Th2 immune reactivity pattern with low production of interferon gamma and high production of interleukin 4 after allergen stimulation of T lymphocytes. Insulin-dependent diabetes mellitus (IDDM) seems to be associated with a Th1 immune reactivity pattern. We therefore postulated that these diseases may be inversely associated. METHODS: We designed a case-control study including 920 children with IDDM, registered in the Danish Registry for Childhood Diabetes, and a sample of 9732 non-diabetic children registered in the Danish Medical Birth Registry. The children were aged 3-15 years. Information on atopic dermatitis was obtained by questionnaires. FINDINGS: The cumulative incidence of atopic dermatitis up to age 15 years was 13.1% among children with IDDM and 19.8% in non-diabetic children (p<0.0001). Among children who developed IDDM, the incidence of atopic dermatitis was significantly lower than in the controls before onset of IDDM (73 cases in 5314 person-months vs 1375 in 57432 person-months; odds ratio 0.49 [0.39-0.63]). After onset of IDDM, diabetic and non-diabetic groups did not differ in incidence of atopic dermatitis (1.36 [0.89 2.07]). INTERPRETATION: Our findings may be explained by different acquired or inherited reactivity patterns associated with atopic dermatitis (Th2) and IDDM (Th1). The results do not allow us to find out whether early development of atopic dermatitis reduces the risk of IDDM, or a propensity for IDDM reduces the risk of early-onset atopic dermatitis. PMID- 11403812 TI - Diagnosis of acute bacterial meningitis in children at a district hospital in sub Saharan Africa. AB - BACKGROUND: The diagnosis of acute bacterial meningitis in children is difficult in sub-Saharan Africa, because the clinical features overlap with those of other common diseases, and laboratory facilities are inadequate in many areas. We have assessed the value of non-laboratory tests and incomplete laboratory data in diagnosing childhood acute bacterial meningitis in this setting. METHODS: We prospectively studied 905 children undergoing lumbar puncture at a rural district hospital in Kenya over 1 year. We related microbiological findings and cerebrospinal-fluid (CSF) laboratory measurements to tests that would typically be available at such a hospital. FINDINGS: Acute bacterial meningitis was proven in 45 children (5.0% [95% CI 3.7-6.6]) and probable in 26 (2.9% [1.9-4.2]). 21 of the 71 cases of proven or probable acute bacterial meningitis had neither neck stiffness nor turbid CSF. In eight of 45 children with proven disease the CSF leucocyte count was less than 10x10(6)/L or leucocyte counting was not possible because of blood-staining. The presence of either a leucocyte count of 50x10(6)/L or more or a CSF/blood glucose ratio of 0.10 or less detected all but two of the 45 children with proven acute bacterial meningitis; these two samples were grossly blood-stained. INTERPRETATION: The diagnosis of childhood acute bacterial meningitis is likely to be missed in a third of cases at district hospitals in sub-Saharan Africa without adequate and reliable laboratory resources. CSF culture facilities are expensive and difficult to maintain, and greater gains could be achieved with facilities for accurate leucocyte counting and glucose measurement. PMID- 11403814 TI - CC chemokine receptor 5 and renal-transplant survival. AB - BACKGROUND: About 1% of white populations are homozygous carriers of an allele of the gene for the CC chemokine receptor 5 (CCR5) with a 32 bp deletion (CCR5Delta32), which leads to an inactive receptor. During acute and chronic transplant rejection, ligands for CCR5 are upregulated, and the graft is infiltrated by CCR5-positive mononuclear cells. We therefore investigated the influence of CCR5Delta32 on renal-transplant survival. METHODS: Genomic DNA from peripheral-blood leucocytes of 1227 renal-transplant recipients was screened by PCR for the presence of CCR5Delta32. Demographic and clinical data were extracted from hospital records. Complete follow-up data were available for 576 recipients of first renal transplants. Graft survival was analysed by Fisher's exact test and Kaplan-Meier plots compared with a log-rank test. FINDINGS: PCR identified 21 patients homozygous for CCR5Delta32 (frequency 1.7%). One patient died with a functioning graft. Only one of the remaining patients lost transplant function during follow-up (median 7.2 years) compared with 78 of the 555 patients with a homozygous wild-type or heterozygous CCR5Delta32 genotype. Graft survival was significantly longer in the homozygous CCR5Delta32 group than in the control group (log-rank p=0.033; hazard ratio 0.367 [95% CI 0.157-0.859]). INTERPRETATION: Patients homozygous for CCR5Delta32 show longer survival of renal transplants than those with other genotypes, suggesting a pathophysiological role for CCR5 in transplant loss. This receptor may be a useful target for the prevention of transplant loss. PMID- 11403815 TI - A swollen right leg. PMID- 11403816 TI - Effect of sensory discrimination training on cortical reorganisation and phantom limb pain. AB - Phantom limb pain is a frequent consequence of the amputation of a body part. Based on the finding that phantom limb pain is closely associated with plastic changes in the primary somatosensory cortex and animal data showing that behaviourally relevant training alters the cortical map, we devised a sensory discrimination training programme for patients with intractable phantom limb pain. Compared with a control group of medically treated patients, the training group had significant reductions in phantom limb pain (p=0.002) and cortical reorganisation (p=0.05) that were positively associated with improved sensory discrimination ability. PMID- 11403817 TI - Fatty fish consumption and risk of prostate cancer. AB - Consumption of fatty fish might reduce the risk of prostate cancer, although epidemiological studies of fish consumption are rare. We studied the association between fish consumption and prostate cancer in a population-based prospective cohort of 6272 Swedish men. During 30 years of follow-up, men who ate no fish had a two-fold to three-fold higher frequency of prostate cancer than those who ate moderate or high amounts did. Our results suggest that fish consumption could be associated with decreased risk of prostate cancer. PMID- 11403818 TI - Congestive heart failure associated with itraconazole. AB - Itraconazole is a synthetic antifungal agent approved in the USA for the treatment of onychomycosis and serious systemic fungal infections. Animal and clinical pharmacology studies suggest negative inotropic effects with itraconazole. Data from the US Food and Drug Administration's Adverse Event Reporting System suggest that use of itraconazole is associated with congestive heart failure. We summarise the details of 58 cases suggestive of congestive heart failure in association with the use of itraconazole. Labelling of itraconazole has been changed to alert physicians to this new finding. PMID- 11403819 TI - Effect of nifedipine in metastatic colon cancer with DNA mismatch repair gene defect. AB - Cancer cells that have DNA mismatch repair deficiency are resistant to many cytotoxic drugs. Calcium channel blockers may inhibit the pathways that cause such resistance. We report a patient with hereditary non-polyposis coli and metastatic colon cancer who had a complete response after treatment with a high dose of nifedipine, a calcium channel blocker. Our findings suggest that drugs that interfere with signal transduction could have a clinical role and deserve further study in selected patients. PMID- 11403820 TI - US physicians and WHO plead for aid to Afghanistan. PMID- 11403821 TI - Natural selection may reduce susceptibility to AIDS. PMID- 11403822 TI - TRAIL leads to apoptosis in acute promyelocytic leukaemia. PMID- 11403826 TI - A century of the US Army yellow fever research. PMID- 11403827 TI - The Bush White House: science advice still out in the cold. PMID- 11403828 TI - Japanese leprosy patients receive official apology. PMID- 11403831 TI - Australian nuclear test veterans produce evidence for compensation. PMID- 11403832 TI - US FDA weighs options for warning on antihypertensive drug. PMID- 11403833 TI - India's trade in fake drugs--bringing the counterfeiters to book. PMID- 11403834 TI - An overview of the immune system. AB - We are continually exposed to organisms that are inhaled, swallowed, or inhabit our skin and mucous membranes. Whether these organisms penetrate and cause disease is a result of both the pathogenicity of the organism (the virulence factors at its disposal) and the integrity of host defence mechanisms. The immune system is an interactive network of lymphoid organs, cells, humoral factors, and cytokines. The essential function of the immune system in host defence is best illustrated when it goes wrong; underactivity resulting in the severe infections and tumours of immunodeficiency, overactivity in allergic and autoimmune disease. In this review we have covered the normal function of the immune system in recognising, repelling, and eradicating pathogens and other foreign molecules. PMID- 11403835 TI - Are disease prevention initiatives working? PMID- 11403836 TI - Graves' disease. PMID- 11403842 TI - The sorrows of young ED. PMID- 11403843 TI - Far-Western based protein-protein interaction screening of high-density protein filter arrays. AB - Even though a rough sketch of the human genome is now available and the number of newly discovered genes, which carry the potential of being biologically and medically relevant is currently greater than ever, only a small proportion has been assigned a biological function. Therefore, enormous attention is now increasingly being drawn towards functional genomics, i.e. the functional characterization of these newly identified sequences. In order to elucidate the role of a particular gene product within its cellular context, we have screened high-density protein filter arrays for protein-protein interactions on the basis of a 'Far-Western' based approach. The methodology described herein easily allows the identification and isolation of cDNAs of proteins, which interact with specific ligands (interacting proteins, antibodies and DNA/RNA sequences), and represents an alternative to tedious conventional protein interaction analyses. Far-Western screening in the context of a whole-genome expression analysis not only facilitates the assignment of biological functions to specific, newly identified protein and DNA sequences, but also is useful in studies that assess the binding capacity of mutant proteins to their interaction partner and in the identification of domains and amino acids involved in known protein-protein interactions. Taken together, we describe an approach that allows the easy and reproducible identification of protein ligands on the basis of a whole-genome expression analysis. PMID- 11403844 TI - Some observations in freeze-drying of recombinant bioluminescent Escherichia coli for toxicity monitoring. AB - A recombinant bioluminescent bacteria, containing a fabA::luxCDABE fusion gene, has been used to characterize freeze-drying methods, which may be conveniently used as a tool for the development of a portable biosensor. Through residual water, viability, biosensing activity and scanning electron microscopy analyses, the characteristics that four cryoprotectants, trehalose, sucrose, sorbitol, and mannitol, conferred on freeze-dried samples were elucidated, including the morphology, water content and activity of the cells. It was found that trehalose showed the best freeze-drying efficiency among the tested cryoprotectants and it might have a specific capacity limitation in protection of the cells during the freeze step. Humidity might result in damage to the cells, according to the viability, when exposed to air during storage, while the water remaining post freeze-drying showed good correlation with damage to the freeze-dried cells when under air-tight storage conditions. The results with other recombinant bioluminescent bacteria indicated that these findings might be general features of the freeze-drying processes. PMID- 11403845 TI - Nucleic acid quantification by delta pH measurement. AB - Nucleic acids are quantitated by UV absorbance measurement, fluorimetry, or hybridization. While the latter method is time-consuming and requires exact knowledge of the sequence, spectroscopic methods require that the sample does not contain UV-absorbing or fluorescent material. An enzymatic method is the measurement of the hyperchromic change upon cleavage of the nucleic acids by nucleases (Kunitz assay). A variation of this assay makes use of the acidification of the solution upon cleavage. We demonstrate here that microgram nucleic acid quantities can be determined when one employs highly active nonspecific nucleases in conjunction with an instrumental setup consisting of a temperature-controlled mixing chamber and miniaturized pH electrodes. Because this method determines the total amount of phosphodiester bonds cleaved, it is independent of the composition or the secondary structure of the nucleic acid and, under certain precautions, represents a simple and robust alternative to optical assays for the determination of either the total nucleic acid concentration or the activity of nucleases in biochemical samples. PMID- 11403846 TI - Antigen presentation using novel particulate organelles from halophilic archaea. AB - A presentation vehicle was developed based on particulate gas vesicles produced by halophilic archaea. Gas vesicle epitope displays were prepared using standard coupling methods or recombinant DNA technology. When presented in the context of gas vesicle preparations, either the hapten, TNP, or a model six amino acid recombinant insert in the outer gas vesicle protein, GvpC was rendered immunogenic. Assays to quantify humoral responses indicated that each preparation elicited strong antibody responses in the absence of exogenous adjuvant. Thus, each preparation elicited a humoral response when injected into mice and this response was long lived and exhibited immunologic memory. Recombinant gas vesicle preparations therefore constitute a new, self-adjuvanting carrier/display vehicle for presentation of an array of peptidyl epitopes. PMID- 11403847 TI - Immobilized salt-tolerant yeasts: application of a new polyethylene-oxide support in a continuous stirred-tank reactor for flavour production. AB - Immobilization of salt-tolerant yeasts considerably decreases the total time required for the flavour development in soy-sauce processes. For immobilization of cells, alginate gel is mostly used as support material. However, alginate is not very suitable for use in soy-sauce processes because alginate is sensitive to abrasion and chemically unstable towards the high salt content of the soy-sauce medium. In contrast, a newly developed polyethylene-oxide gel seems to be more suitable, but this gel has not been used so far for flavour production in a bioreactor with a high salt content. Therefore, this gel was applied with immobilized salt-tolerant yeasts in a continuous stirred-tank reactor, containing more than 12.5% (w/v) salt. In this reactor, the polyethylene-oxide gel particles did not show any abrasion for several days, while alginate gel beads were already destroyed within 1 day. In addition, the polyethylene-oxide gel particles with immobilized salt-tolerant yeasts Candida versatilis and Zygosaccharomyces rouxii showed a good flavour production. From this work, it was concluded that the application of polyethylene-oxide gel in long-term soy-sauce processes is attractive in the case the sticking together of polyethylene-oxide gel particles can be controlled. PMID- 11403848 TI - Long and accurate PCR with a mixture of KOD DNA polymerase and its exonuclease deficient mutant enzyme. AB - DNA polymerase from Thermococcus kodakaraensis KOD1 (previously Pyrococcus sp. KOD1) is one of the most efficient thermostable PCR enzymes exhibiting higher accuracy and elongation velocity than any other commercially available DNA polymerase [M. Takagi et al. (1997) Appl. Environ. Microbiol. 63, 4504-4510]. However, when long distance PCR (>5 kbp) was performed with KOD DNA polymerase, amplification efficiency (product yield) becomes lower because of its strong 3' 5' exonuclease activity for proof-reading. In order to improve a target length limitation in PCR, mutant DNA polymerases with decreased 3'-5' exonuclease activity were designed by substituting amino acid residues in conserved exonuclease motifs, Exo I (Asp141-Xaa-Glu), Exo II (Asn210-Xaa-Xaa-Xaa-Phe-Asp), and Exo III (Tyr311-Xaa-Xaa-Xaa-Asp). Exonuclease activity and amplification fidelity (error rate) of the DNA polymerases were altered by mutagenesis. However, long and accurate PCR by a single-type of mutant DNA polymerase was very difficult. The wild-type DNA polymerase (WT) and its exonuclease deficient mutant (N210D) were mixed in different ratio and their characteristics in PCR were examined. When the mixed enzyme (WT and N210D) was made at the ratio of 1:40, long PCR (15 kbp) at lower mutation frequency could be efficiently achieved. PMID- 11403849 TI - The Zygosaccharomyces rouxii strain CBS732 contains only one copy of the HOG1 and the SOD2 genes. AB - The osmotolerant yeast Zygosaccharomyces rouxii CBS732 contains only one copy of the ZrHOG1 and ZrSOD2-22 genes. Both genes were cloned and sequenced (Acc. Nos. AJ132606 and AJ252273, respectively) and their sequences were compared to homologous pairs of genes from Z. rouxii ATCC42981 (genes Z-HOG1, Z-HOG2, Z-SOD2, Z-SOD22). The CBS732 ZrHog1p is shorter than its ATCC42981 counterparts (380 aa residues vs. 407 and 420 aa, respectively) and is more similar to ATCC42981 Z Hog2p than to Z-Hog1p. Also its promoter region corresponds to that one of Z HOG2. The CBS732 ZrHOG1 promoter region is recognised by Saccharomyces cerevisiae, and the gene product (MAP kinase ZrHog1p) presence fully complements the osmosensitivity of a S. cerevisiae hog1 mutant strain. The CBS ZrSOD2-22 gene is highly similar to ATCC42981 Z-SOD2 but it contains also a segment of 15 aa residues specific for Z-SOD22. Z. rouxii ZrSod2-22 Na(+)/H(+) antiporter expressed in S. cerevisiae shows better activity toward toxic Na(+) and Li(+) cations than does S. cerevisiae's own Nha1 antiporter, and is efficient in improving the halotolerance of some S. cerevisiae wild types. PMID- 11403850 TI - An efficient gene transfer system for hematopoietic cell line using transient and stable vectors. AB - The hematopoietic system represents an interesting model for gene transfer protocols. Here, we have evaluated the efficiency of a gene transfer system using the polycationic compound SuperFect (Qiagen) and the K562 hematopoietic cell line. Transient and stable vectors carrying the enhanced green fluorescent protein (EGFP) reporter gene were employed. The stable vector was constructed based on Epstein-Barr virus sequences such as EBV oriP (origin of replication) and EBNA (EBV nuclear antigen)-1, both for DNA replication. The transfection efficiency of the viable cells was estimated by flow cytometry at approximately 98% for transient and stable vectors. Transiently transfected cells presented optimal EGFP expression until day 2 when fluorescence started to decrease. In contrast, stable transfectants continuously expressed the marker gene product for 10 weeks in the presence of G418. Our results represent an efficient gene transfer method for K562 hematopoietic cells and may be used as an alternative approach for further gene transfer studies involving hematopoietic cells. PMID- 11403851 TI - A P450 BM-3 mutant hydroxylates alkanes, cycloalkanes, arenes and heteroarenes. AB - P450 monooxygenases from microorganisms, similar to those of eukaryotic mitochondria, display a rather narrow substrate specificity. For native P450 BM 3, no other substrates than fatty acids or an indolyl-fatty acid derivative have been reported (Li, Q.S., Schwaneberg, U., Fischer, P., Schmid, R.D., 2000. Directed evolution of the fatty-acid hydroxylase P450BM-3 into an indole hydroxylating catalyst. Chem. Eur. J. 6 (9), 1531-1536). Engineering the substrate specificity of Bacillus megaterium cytochrome P-450 BM3: hydroxylation of alkyl trimethylammonium compounds. Biochem. J. 327, 537-544). We thus were quite surprised to observe, in the course of our investigations on the rational evolution of this enzyme towards mutants, capable of hydroxylating shorter-chain fatty acids, that a triple mutant P450 BM-3 (Phe87Val, Leu188-Gln, Ala74Gly, BM-3 mutant) could efficiently hydroxylate indole, leading to the formation of indigo and indirubin (Li, Q.S., Schwaneberg, U., Fischer, P., Schmid, R.D., 2000. Directed evolution of the fatty-acid hydroxylase P450BM-3 into an indole hydroxylating catalyst. Chem. Eur. J. 6 (9), 1531-1536). Indole is not oxidized by the wild-type enzyme; it lacks the carboxylate group by which the proper fatty acid substrates are supposed to be bound at the active site of the native enzyme, via hydrogen bonds to the charged amino acid residues Arg47 and Tyr51. Our attempts to predict the putative binding mode of indole to P450 BM-3 or the triple mutant by molecular dynamics simulations did not provide any useful clue. Encouraged by the unexpected activity of the triple mutant towards indole, we investigated in a preliminary, but systematic manner several alkanes, alicyclic, aromatic, and heterocyclic compounds, all of which are unaffected by the native enzyme, for their potential as substrates. We here report that this triple mutant indeed is capable to hydroxylate a respectable range of other substrates, all of which bear little or no resemblance to the fatty acid substrates of the native enzyme. PMID- 11403852 TI - The effect of agitation and nitrogen concentration on lignin peroxidase (LiP) isoform composition during fermentation of Phanerochaete chrysosporium. AB - Convective Interaction Media (CIM) monolithic columns were applied for the HPLC monitoring of Phanerochaete chrysosporium lignin peroxidase (LiP) isoforms during cultivation. The influence of the agitation mode (circular, elliptic) and rate (130 and 200 rpm), as well as the initial nitrogen concentration (1.6-6 mM) in the growth medium was investigated. Identical rotation rate but different agitation modes resulted in different LiP activities and isoenzyme compositions. On the other hand, at different agitation types and rates, similar LiP activities were obtained at different isoenzyme compositions. Although LiP H2 and LiP H6/H7 were predominant isoenzymes obtained at various cultivation conditions, relative isoenzyme amounts differ considerably when initial nitrogen concentration was changed between 1.6 and 5 mM. PMID- 11403853 TI - The effect of shaking regime on the rate and extent of enzymatic hydrolysis of cellulose. AB - In an attempt to elucidate the effect of mixing on the rate and extent of enzymatic hydrolysis of cellulosic substrates, alpha-cellulose was hydrolysed using a commercial cellulase preparation at varying levels of substrate concentration (2.5,5 and 7.5% (w/v)) and by using three shaking regimes: continuous at low-speed (25 rpm), continuous at high-speed (150 rpm) and an intermittent regime comprised of high and low-speed shaking intervals. The continuous, high-speed shaking produced the highest conversion yields, whereas the intermittent and low-speed shaking regimes resulted in lower conversions. After 72 h, at all shaking regimes (150 rpm,25 rpm and intermittent), using a low substrate concentration (2.5%) produced conversion yields (82,79 and 80%) higher than those obtained at high (7.5%) substrate concentration (68,63 and 68%). As the substrate concentration increased, the conversion yields at intermittent shaking gradually approached those resulting from high-speed shaking. Thus, it appears that intermittent shaking could be a beneficial process option as it can reduce the mixing energy requirements while producing reasonably high conversion yields. PMID- 11403854 TI - Mapping and promoter sequencing of HNF-1beta gene in diabetes-prone and resistant mice. AB - By using a novel single nucleotide polymorphism (SNP) in the coding sequence, the chromosomal location of Tcf2, encoding hepatic nuclear factor (HNF)-1beta, was determined in F2 intercrosses between Nagoya-Shibata-Yasuda (NSY) mice, an animal model of type 2 diabetes, and control C3H/He mice. The promoter region of Tcf2 gene was sequenced in NSY, non-obese diabetic (NOD) and control C3H/He mice. Tcf2 was mapped between genetic markers D11MIT320 and D11MIT195 with the following distances: D11MIT320-(7.3 cM)-Tcf2-(0.5 cM)-D11MIT195. A variant with insertion of C between -205 and -204 in the promoter region of Tcf2 was identified in NSY mice, but not NOD and C3H/He mice. PMID- 11403855 TI - Insulin action and skeletal muscle blood flow in patients with Type 1 diabetes and microalbuminuria. AB - Our objective was to determine whether Type 1 diabetic patients with microalbuminuria are less sensitive to the effects of insulin on glucose metabolism and skeletal muscle blood flow, compared to those with normal albumin excretion, after careful matching for confounding variables. We recruited 10 normotensive Type 1 diabetic patients with microalbuminuria and 11 with normoalbuminuria matched for age, sex, body mass index, duration of diabetes and HbA(1c). Peripheral and hepatic insulin action was assessed using a two-step euglycaemic hyperinsulinaemic clamp (2 h at 0.4 mU x kg(-1) x min(-1), 2 h at 2.0 mU x kg(-1) x min(-1)) combined with isotope dilution methodology. Skeletal muscle blood flow was determined by venous occlusion plethysmography. During the clamps, glucose infusion rates required to maintain euglycaemia were similar in the microalbuminuric subjects and controls (step 1, 8.2+/-1.4 (SE) vs 9.2+/-1.3 micromol x kg(-1) x min(-1): step 2, 30.9+/-2.7 vs 32.0+/-3.8 micromol x kg(-1) x min(-1)), as was hepatic glucose production basally and at steady state in step 1. In step 2, hepatic glucose production was lower in the microalbuminuric group (2.9+/-0.9 vs 6.4+/-0.7 micromol x kg(-1) x min(-1), P=0.005). During step 2, skeletal muscle blood flow increased significantly above baseline in the normoalbuminuric group (4.1+/-0.5 vs 3.2+/-0.4 ml x 100-ml(-1) x min(-1), P=0.01) but not in the microalbuminuric group (2.4+/-0.3 vs 2.3+/-0.4 ml x 100-ml(-1) x min(-1)). In conclusion, microalbuminuria in Type 1 diabetes was found to be associated with impairment of insulin-mediated skeletal muscle blood flow, but not with insulin resistance. PMID- 11403856 TI - Early alterations of blood pressure in normotensive and normoalbuminuric Type 1 diabetic patients. AB - With the objective to examine patterns of blood pressure (BP) in normotensive and normoalbuminuric Type 1 diabetic patients during 24 h ambulatory blood pressure monitoring (ABPM) we studied 28 Type 1 diabetic patients aged 27+/-7.1 years with a disease duration of 9+/-6.6 years, and 28 non-diabetic normotensive subjects aged 25+/-6.5 years matched to the diabetic group for age, gender, skin color, weight, height, body mass index, clinic BP and absence of microalbuminuria. Systolic BP (sBP) and diastolic BP (dBP) were recorded for 24 h, daytime and nighttime. SBP and dBP burden, night/day BP ratios and percent nighttime BP fall were determined. Subjects with a nocturnal fall in either sBP or dBP of less than 10% of daytime values were classified as non-dippers. Both sBP (111+/-7.1 vs. 104+/-9 mmHg; P=0.003) and dBP nighttime (66+/-6.1 vs. 61+/-5.3 mmHg; P=0.001) were higher in diabetic patients than non-diabetic subjects. Night/day ratios for sBP (0.93+/-0.04 vs. 0.89+/-0.05; P=0.006) and dBP (0.86+/-0.06 vs. 0.82+/-0.06; P=0.007) were higher in diabetics. The loss of a fall in sBP was more prevalent in diabetic subjects (78 vs. 39%; P=0.007). Non-dippers for sBP and dBP in the diabetic group had higher BP burden during the nighttime (21.4+/-16.6 vs. 3.2+/ 3.9%; P=0.01 and 21.9+/-10 vs. 3.7+/-5.5%; P<0.001, respectively). Our data demonstrate higher sBP and dBP during the nighttime and loss of the nocturnal fall in BP in Type 1 diabetic patients. Further prospective studies are needed to define if high BP burden in diabetic non-dippers during the night could represent a risk for nephropathy and cardiovascular disease. PMID- 11403857 TI - Importance of standardization of hemoglobin A1c in the analysis of factors that predict hemoglobin A1c levels in non-diabetic residents of three distinct areas of Japan. AB - We performed a statistical analysis to elucidate effects of standardized measurement of hemoglobin A1c (HbA1c) on analysis of factors that affect HbA1c values. Subjects were participants in the Japan Public Health Center-based Prospective Study on Cancer and Cardiovascular Diseases, and a total of 1789 men and 3150 women in three distinct areas who did not have overt diabetes (HbA1c> or =6.1% or prior diagnosis) were analyzed. A different method of HbA1c assay was used in each area: high-performance liquid chromatography in one area and a different immunochemical method in each of the other two areas. Then, calibration of HbA1c was performed using two HbA1c standards (5.5 and 10.5%) provided by the Japan Diabetes Society. Analysis of co-variance was performed separately in men and women. When raw HbA1c data were used as the outcome, 'area', which represents differences in assay systems, lifestyles, etc. had a significant effect on HbA1c levels. When calibrated HbA1c data were used, however, 'area' was no longer a significant factor. In the latter analysis, age and BMI were the principal contributors to HbA1c, and parental history of diabetes had a weak effect in women. Thus, standardization of HbA1c reduced the difference between assay systems, and uncovered two common factors to determine HbA1c levels. PMID- 11403858 TI - Insulin resistance precedes the appearance of albuminuria in non-diabetic subjects: 6 years follow up study. AB - Microalbuminuria is a marker of increased risk of cardiovascular mortality in type 1 and type 2 diabetes, and in non-diabetic subjects. Little is known about the association between prospective changes of microalbuminuria and the risk factors of atherosclerosis, or gene polymorphism in non-diabetic subjects. We conducted a 6-year prospective study of risk factors for progression of albuminuria in non-diabetic subjects. The participants were 116 non-diabetic subjects who consecutively underwent medical examinations for Japanese-Americans living in Hawaii. In the baseline examination in 1992, normoalbuminuria was found in all subjects. After 6 years, 101 subjects remained normoalbuminuria (non progressors), 15 subjects changing from normoalbuminuria to microalbuminuria or proteinuria and were defined as progressors. In progressors, compared with non progressors, the fasting insulin level and HOMA-R were significantly higher at 3 years follow-up, and the systolic and diastolic pressure and Sigma insulin level were significantly higher at 6 years follow-up. Insulin resistance appeared earlier than the appearance of hypertension and albuminuria. In progressors, there was no significant correlation with angiotensin-converting enzyme (ACE) genotype or angiotensinogen (AGT) genotype compared with non-progressors. Therefore, the appearance of insulin resistance should be regarded as a remarkable mediator of albuminuria. PMID- 11403859 TI - The Poole Diabetes Study: how many cases of Type 2 diabetes are diagnosed each year during normal health care in a defined community? AB - We have investigated the incidence of newly diagnosed Type 2 diabetes in the Poole area and extrapolated it to the rest of the UK. METHODS: this prospective observational study used a surveillance programme in primary and secondary care. We identified all cases of newly diagnosed Type 2 diabetes mellitus occurring from 1st May 1996 to 30th June 1998 through the normal health care process without any active screening in 186889 people registered with 24 primary care practices in the Poole area. RESULTS: the 1996 prevalence of diagnosed Type 2 diabetes in this population was 1.59 (95% CI 1.53-1.65%)%. During the first 24 months of the study, 706 new cases of Type 2 diabetes mellitus, 382 men and 324 women, were identified. The crude annual incidence of newly diagnosed Type 2 diabetes, thus was 1.93/1000 (95% CI 1.73-2.13%) and age/sex adjusted incidence was 1.67/1000 (95% CI 1.49-1.84%). The age-adjusted incidence was higher in men, 1.86/1000 (95% CI 1.60-2.13), than in women, 1.48/1000 (95% CI 1.25-1.71%), relative risk 1.26 (95% CI 0.997-1.527%), but this difference did not reach statistical significance. Mean HbA1c at diagnosis was 10.8 (S.D. 2.9%)%. Men were younger at diagnosis than women (mean age, 62.9 vs. 65.9%, P<0.01). CONCLUSION: in UK, prior to the change in the WHO diagnostic criteria for diabetes, we estimate that over 98000 new cases of Type 2 diabetes were diagnosed each year. PMID- 11403860 TI - Predictors of mortality from type 2 diabetes mellitus in Canterbury, New Zealand; a ten-year cohort study. AB - The aim was to establish mortality rates in a cohort of subjects with type 2 diabetes mellitus over 10 years in Canterbury, New Zealand (NZ) and to determine baseline prognostic factors. Subjects (447) with type 2 diabetes (208 male, 239 female; age range 30-82 years, median 62 years; of predominantly European origin) were characterised in a clinic survey in 1989. Individual status (dead or alive) at June 1 1999 (10 year follow-up) was ascertained. Mortality rates were compared with the general NZ population and the relative risk (RR) of baseline prognostic factors evaluated with Cox's proportional hazards model. At 10 years, 232 subjects were confirmed as alive and 187 as dead - only 28 were untraceable. Ten year survival was 55% (95% CI: 50-60) for the cohort, compared with 70% (95% CI: 65-75) at 6 years. Factors assessed at baseline (1989), that were independently prognostic of total mortality, included age (RR 2.0, 95% CI: 1.6-2.5), pre existing coronary artery disease (CAD; RR 1.7, 95% CI: 1.2-2.4) and albuminuria (RR 1.58, 95% CI: 1.1-2.3). Glycated haemoglobin was not a significant predictor of total mortality, although was a predictor of CAD mortality in those subjects free of CAD in 1989 (RR 1.6, 95% CI: 1.1-2.3). In the latter subset, independent prognostic factors for CAD mortality also included age (RR 2.5, 95% CI: 1.7-3.8), hypertension (RR 1.9, 95% CI: 1.0-3.7), peripheral vascular disease (RR 2.4, 95% CI: 1.3-4.5) and smoking (RR 2.6, 95% CI: 1.2-5.8). Increased mortality in type 2 diabetic subjects is therefore attributable to multiple risk factors. Improved outcomes will depend on interventions targeted at glycaemic and all other remediable factors. PMID- 11403861 TI - Clinical and technical evaluation of a new self-monitoring blood glucose meter: assessment of analytical and user error. AB - Our study aimed to validate (against the standards of the American Diabetes Association and the International Organization for Standardization) the analytical and clinical accuracy of the new MediSense Precision Plus Electrodes on the QID system when compared with a reference method using the Abbott Vision glucose analyzer. Previous studies have shown that the overall accuracy of the device also depends on the proficiency of the operator, so we also assessed the 'ease of use' of the MediSense system, by comparing the results obtained by the patient and health care professional. Accuracy of the self-monitored blood glucose measurements was evaluated over a wide range of glucose readings (2.6 20.0 mmol/l). Between-run CVs (using the manufacturer's quality control material) were found to be 7% at 2.7 mmol/l and 4.8% at 15.5 mmol/l (n=380). We used the error-grid analysis with target range blood glucose, then separated the data into different subsets. We found that 100% of all measurements were in the clinically acceptable zones of A and B. All measured values of the MediSense QID system complied with the requirements for 'blood glucose monitoring meters', as proposed by the International Organization for Standardization. The rating of the patient questionnaire showed a good to very good overall rating and acceptance with a short instruction time. The results indicate that that the MediSense QID/Precision Plus Electrodes is a reliable and easy to use device, which can be recommended for the majority of patients with diabetes mellitus. PMID- 11403863 TI - Speciation. PMID- 11403862 TI - Comparison of peripheral arterial reconstruction in diabetic and non-diabetic patients: a prospective clinic-based study. AB - To assess the efficacy and safety of lower extremity arterial reconstruction in diabetic and non-diabetic subjects during a 3-year period. A prospective clinic based study between 1994-1999 in Area 7, Madrid, with a population of 569307 and an estimated diabetic population of 37932 (15505 men and 22427 women). The level of arterial reconstruction and associated risk factors were ascertained. RESULTS: A total of 588 peripheral revascularization surgical procedures were performed in 481 patients. The diabetic patients (n=174, 36.2%) underwent 222 surgical procedures (including 48 follow-on operations, 21.6%), and 307 non-diabetic subjects underwent 366 surgical procedures (59 follow-on operations, 16.1%). The numbers of surgical procedures per 100000 people at risk and year were 18.8 and 1.8 for non-diabetic men and women, respectively, and 145.1 and 29.0 for men and women with diabetes mellitus (7.7- and 16.2-fold, respectively). Age at reconstruction surgery was 2 and 5 years earlier in non-diabetic than in diabetic men and women, respectively. Diabetic patients had a higher neuropathy score (P<0.05) and were less frequently smokers (P<0.05) than non-diabetic subjects. Diabetic subjects more frequently had distal reconstruction while proximal arterial reconstruction was more often performed in non-diabetic subjects. Between 64.6 and 80.4% of people with diabetes and 82.3 and 88.9% of non-diabetic subjects had no complications during their in-hospital stay. Distal amputation simultaneous to arterial reconstruction was the most frequent morbidity of people with diabetes during the study (P<0.05). Despite a graft occlusion rate after femoropopliteal revascularization significantly higher than in non-diabetic people (P<0.05), diabetic people more often required lower extremity amputations (LEAs) for the same level of bypass (P<0.01). Cumulative limb salvage rates were lower in diabetic patients than in non-diabetic subjects at femoropopliteal (49.2 vs. 89.7%; P<0.001), femorodistal (73.5 vs. 95.2%; P<0.01), and distal reverse (77.9 vs. 87.3%; P<0.05) arterial reconstruction, at the end of the third year, but similar after aorto-iliac reconstruction (93.1 vs. 97.5%). A higher neuropathy score and the presence of foot ulcers were associated to significantly lower limb salvage in diabetic patients (P<0.05), but not in non-diabetic people. Survival rates after 3 years were similar between diabetic and non-diabetic populations after aorto-iliac (93.1 vs. 97.5%), femoropopliteal (97.2 vs. 90.3%), and distal reverse (93.2 vs. 98.1%) revascularization, and slightly lower in diabetic compared to non-diabetic patients after femorodistal revascularization (82.1 vs. 96.3%; P<0.05). CONCLUSION: Although limb salvage after arterial reconstruction is lower in diabetic than in non-diabetic subjects, particularly in those with a higher neuropathy score, this surgical approach can be applied in both diabetic and non-diabetic subjects with otherwise similar outcome. PMID- 11403864 TI - The mind of the species problem. AB - The species problem is the long-standing failure of biologists to agree on how we should identify species and how we should define the word 'species'. The innumerable attacks on the problem have turned the often-repeated question 'what are species?' into a philosophical conundrum. Today, the preferred form of attack is the well-crafted argument, and debaters seem to have stopped inquiring about what new information is needed to solve the problem. However, our knowledge is not complete and we have overlooked something. The species problem can be overcome if we understand our own role, as conflicted investigators, in causing the problem. PMID- 11403865 TI - Theory and speciation. AB - The study of speciation has become one of the most active areas of evolutionary biology, and substantial progress has been made in documenting and understanding phenomena ranging from sympatric speciation and reinforcement to the evolutionary genetics of postzygotic isolation. This progress has been driven largely by empirical results, and most useful theoretical work has concentrated on making sense of empirical patterns. Given the complexity of speciation, mathematical theory is subordinate to verbal theory and generalizations about data. Nevertheless, mathematical theory can provide a useful classification of verbal theories; can help determine the biological plausibility of verbal theories; can determine whether alternative mechanisms of speciation are consistent with empirical patterns; and can occasionally provide predictions that go beyond empirical generalizations. We discuss recent examples of progress in each of these areas. PMID- 11403866 TI - The genetics of species differences. AB - Species are separated by reproductive isolation as well as by more 'ordinary' differences in morphology and behavior that play no necessary role in blocking gene flow. Although a great deal is now known about the genetics of reproductive isolation, we are only beginning to understand the genetic basis of ordinary phenotypic differences between species. I review what is known about the number of genes involved in such differences, as well as about the role of major genes and epistasis in the evolution of these differences. I also compare and contrast these findings with those on the genetics of reproductive isolation. PMID- 11403867 TI - Chromosomal rearrangements and speciation. AB - Several authors have proposed that speciation frequently occurs when a population becomes fixed for one or more chromosomal rearrangements that reduce fitness when they are heterozygous. This hypothesis has little theoretical support because mutations that cause a large reduction in fitness can be fixed through drift only in small, inbred populations. Moreover, the effects of chromosomal rearrangements on fitness are unpredictable and vary significantly between plants and animals. I argue that rearrangements reduce gene flow more by suppressing recombination and extending the effects of linked isolation genes than by reducing fitness. This unorthodox perspective has significant implications for speciation models and for the outcomes of contact between neospecies and their progenitor(s). PMID- 11403868 TI - Gene trees and species trees are not the same. AB - The relationship between species is usually represented as a bifurcating tree with the branching points representing speciation events. The ancestry of genes taken from these species can also be represented as a tree, with the branching points representing ancestral genes. The time back to the branching points, and even the branching order, can be different between the two trees. This possibility is widely recognized, but the discrepancies are often thought to be small. A different picture is emerging from new empirical evidence, particularly that based on multiple loci or on surveys with a wide geographical scope. The discrepancies must be taken into account when estimating the timing of speciation events, especially the more recent branches. On the positive side, the different timings at different loci provide information about the ancestral populations. PMID- 11403869 TI - Sexual selection and speciation. AB - The power of sexual selection to drive changes in mate recognition traits gives it the potential to be a potent force in speciation. Much of the evidence to support this possibility comes from comparative studies that examine differences in the number of species between clades that apparently differ in the intensity of sexual selection. We argue that more detailed studies are needed, examining extinction rates and other sources of variation in species richness. Typically, investigations of extant natural populations have been too indirect to convincingly conclude speciation by sexual selection. Recent empirical work, however, is beginning to take a more direct approach and rule out confounding variables. PMID- 11403870 TI - Ecology and the origin of species. AB - The ecological hypothesis of speciation is that reproductive isolation evolves ultimately as a consequence of divergent natural selection on traits between environments. Ecological speciation is general and might occur in allopatry or sympatry, involve many agents of natural selection, and result from a combination of adaptive processes. The main difficulty of the ecological hypothesis has been the scarcity of examples from nature, but several potential cases have recently emerged. I review the mechanisms that give rise to new species by divergent selection, compare ecological speciation with its alternatives, summarize recent tests in nature, and highlight areas requiring research. PMID- 11403871 TI - Sympatric speciation in animals: the ugly duckling grows up. AB - Sympatric speciation has become increasingly accepted in the past decade, as a result of new models substantiating its plausibility and new evidence that the conditions specified by the models are met in many natural populations. Retrospective phylogenetic and population genetic signatures of sympatric speciation have also been derived, and these are beginning to be tested. This new work has helped increase the acceptance of sympatric speciation as a plausible process, although it remains difficult to show conclusively that specific pairs of taxa have speciated through sympatric processes alone. It might be time for a re-evaluation of the geographical classification of speciation modes in favor of one based primarily on evolutionary mechanisms PMID- 11403872 TI - Phylogenetics and speciation. AB - Species-level phylogenies derived from molecular data provide an indirect record of the speciation events that have led to extant species. This offers enormous potential for investigating the general causes and rates of speciation within clades. To make the most of this potential, we should ideally sample all the species in a higher group, such as a genus, ensure that those species reflect evolutionary entities within the group, and rule out the effects of other processes, such as extinction, as explanations for observed patterns. We discuss recent practical and theoretical advances in this area and outline how future work should benefit from incorporating data from genealogical and phylogeographical scales. PMID- 11403873 TI - Scale and species numbers. AB - One of the main tasks confronting community ecologists is to explain why a particular site harbours a certain number of species. The site might range from a drop of water to the whole Earth, and the species might be drawn from a very restricted taxon or include all living organisms. The common problem, however, is to understand the relative importance of speciation and extinction and, more locally, of immigration and loss. Speciation is the ultimate motor driving biodiversity and ecologists need to know the factors influencing rates of speciation, and whether there is a feedback, positive or negative, between species numbers and the generation of new taxa. However, the relative importance of speciation and other factors determining species numbers varies crucially across different scales of enquiry. Here, we explore some of these issues as we move from a macro- to microscale perspective, focusing on a limited number of studies that we believe make important advances in the field. PMID- 11403874 TI - Speciation in the fossil record. AB - It is easy to claim that the fossil record says nothing about speciation because the biological species concept (which relies on interbreeding) cannot be applied to it and genetic studies cannot be carried out on it. However, fossilized organisms are often preserved in sufficient abundance for populations of intergrading morphs to be recognized, which, by analogy with modern populations, are probably biological species. Moreover, the fossil record is our only reliable documentation of the sequence of past events over long time intervals: the processes of speciation are generally too slow to be observed directly, and permanent reproductive isolation can only be verified with hindsight. Recent work has shown that some parts of the fossil record are astonishingly complete and well documented, and patterns of lineage splitting can be examined in detail. Marine plankton appear to show gradual speciation, with subsequent morphological differentiation of lineages taking up to 500000 years to occur. Marine invertebrates and vertebrates more commonly show punctuated patterns, with periods of rapid speciation followed by long-term stasis of species lineages. PMID- 11403875 TI - Speciation Glossary. PMID- 11403876 TI - P2X receptors in peripheral neurons. AB - P2X receptors are a family of ligand-gated ion channels, activated by extracellular ATP. The seven subunits cloned (P2X1-7) can assemble to form homomeric and heteromeric receptors. Peripheral neurons of neural crest origin (e.g. those in dorsal root, trigeminal, sympathetic and enteric ganglia) and placodal origin (e.g. those in nodose and petrosal ganglia) express mRNAs for multiple P2X subunits. In this review, we summarize the molecular biological, electrophysiological and immunohistochemical evidence for P2X receptor subunits in sensory, sympathetic, parasympathetic, pelvic and myenteric neurons and adrenomedullary chromaffin cells. We consider the pharmacological properties of these native P2X receptors and their physiological roles. The responses of peripheral neurons to ATP show considerable heterogeneity between cells in the same ganglia, between ganglia and between species. Nevertheless, these responses can all be accounted for by the presence of P2X2 and P2X3 subunits, giving rise to varying proportions of homomeric and heteromeric receptors. While dorsal root ganglion neurons express predominantly P2X3 and rat sympathetic neurons express mainly P2X2 receptors, nodose and guinea-pig sympathetic neurons express mixed populations of P2X2 and heteromeric P2X2/3 receptors. P2X receptors are important for synaptic transmission in enteric ganglia, although their roles in sympathetic and parasympathetic ganglia are less clear. Their presence on sensory neurons is essential for some processes including detection of filling of the urinary bladder. The regulation of P2X receptor expression in development and in pathological conditions, along with the interactions between purinergic and other signalling systems, may reveal further physiological roles for P2X receptors in autonomic and sensory ganglia. PMID- 11403877 TI - Molecular pathways involved in the neurotoxicity of 6-OHDA, dopamine and MPTP: contribution to the apoptotic theory in Parkinson's disease. AB - Parkinson's disease (PD) is a neurodegenerative disorder characterized by a preferential loss of the dopaminergic neurons of the substantia nigra pars compacta. Although the etiology of PD is unknown, major biochemical processes such as oxidative stress and mitochondrial inhibition are largely described. However, despite these findings, the actual therapeutics are essentially symptomatical and are not able to block the degenerative process. Recent histological studies performed on brains from PD patients suggest that nigral cell death could be apoptotic. However, since post-mortem studies do not allow precise determination of the sequence of events leading to this apoptotic cell death, the molecular pathways involved in this process have been essentially studied on experimental models reproducing the human disease. These latter are created by using neurotoxic compounds such as 6-hydroxydopamine (6-OHDA), 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or dopamine (DA). Extensive study of these models have shown that they mimick, in vitro and in vivo, the histological and/or the biochemical characteristics of PD and thus help to define important cellular actors of cell death presumably critical for the nigral degeneration. This review reports recent data concerning the biochemical and molecular apoptotic mechanisms underlying the experimental models of PD and correlates them to the phenomena occurring in human disease. PMID- 11403878 TI - Alteration of second messengers during acute cerebral ischemia - adenylate cyclase, cyclic AMP-dependent protein kinase, and cyclic AMP response element binding protein. AB - A variety of neurotransmitters and other chemical substances are released into the extracellular space in the brain in response to acute ischemic stress, and the biological actions of these substances are exclusively mediated by receptor linked second messenger systems. One of the well-known second messenger systems is adenylate cyclase, which catalyzes the generation of cyclic AMP, triggering the activation of cyclic AMP-dependent protein kinase (PKA). PKA controls a number of cellular functions by phosphorylating many substrates, including an important DNA-binding transcription factor, cyclic AMP response element binding protein (CREB). CREB has recently been shown to play an important role in many physiological and pathological conditions, including synaptic plasticity and neuroprotection against various insults, and to constitute a convergence point for many signaling cascades. The autoradiographic method developed in our laboratory enables us to simultaneously quantify alterations of the second messenger system and local cerebral blood flow (lCBF). Adenylate cyclase is diffusely activated in the initial phase of acute ischemia (< or = 30 min), and its activity gradually decreases in the late phase of ischemia (2-6 h). The areas of reduced adenylate cyclase activity strictly coincide with infarct areas, which later become visible. The binding activity of PKA to cyclic AMP, which reflects the functional integrity of the enzyme, is rapidly suppressed during the initial phase of ischemia in the ischemic core, especially in vulnerable regions, such as the CA1 of the hippocampus, and it continues to decline. By contrast, PKA binding activity remains enhanced in the peri-ischemia area. These changes occur in a clearly lCBF-dependent manner. CREB phosphorylation at a serine residue, Ser(133), which suggests the activation of CREB-mediated transcription of genes containing a CRE motif in the nuclei, remains enhanced in the peri-ischemia area, which is spared of infarct damage. On the other hand, CREB phosphorylation at Ser133 rapidly diminishes in the ischemic core before the histological damage becomes manifest. The Ca2+ influx during membrane depolarization contributes to CREB phosphorylation in the initial phase of post-ischemic recirculation, while PKA activation and other signaling elements seem to be responsible in the later phase. These findings suggest that derangement of cyclic AMP-related intracellular signal transduction closely parallels ischemic neuronal damage and that persistent enhancement of this signaling pathway is important for neuronal survival in acute cerebral ischemia. PMID- 11403880 TI - Models of protein crystal growth. AB - The growth of large and well ordered protein crystals remains the major obstacle in protein structure determination by means of X-ray crystallography. One of the reasons is that the physico-chemical aspect of protein crystallization process is not understood. This article reviews efforts towards formulation of models that could become theoretical frameworks for the interpretation of voluminous experimental data collected on protein crystal growth. Special attention is devoted to microscopic models that recognize the role of the shape of protein molecules in crystal formation. PMID- 11403881 TI - Cooperativity of phospholipid reorganization upon interaction of dipyridamole with surface monolayers on water. AB - Results from various surface sensitive characterization techniques suggest a model for the interaction of the piperidinopyrimidine dipyridamole (DIP)--known as a vasodilator and inhibitor of P-glycoprotein associated multidrug resistance of tumor cells--with phospholipid monolayers in which the drug is peripherally associated with the membrane, binding (up to) five phospholipids at a time. These multiple interactions are responsible for a very strong association of the drug with the lipid monolayer even at exceedingly low concentrations (approximately 0.2 mol%). Electrostatic interactions and hydrogen bonding are likely involved in the binding of DIP to DPPC. Cooperative effects among the lipids are invoked to explain the macroscopically measurable changes of lipid monolayer properties even when only one out of 100 DPPC molecules is directly associated with a DIP molecule. A reversal of the observed changes upon drug association with the membrane as the DIP concentration surpasses a threshold concentration (c(crit)approximately 0.5 mol%) may be explained by cooperativity in a different context, the self-aggregation of drug molecules. With its implications for the interaction of DIP with phospholipid films, this work provides a first approach to the explanation of the high sensitivity of cell membranes to piperidinopyrimidine drugs on a molecular level. PMID- 11403882 TI - Chlorpromazine associates with phosphatidylserines to cause an increase in the lipid's own interfacial molecular area--role of the fatty acyl composition. AB - Partition coefficients of the drug chlorpromazine were determined for five different molecular species of diacylglycerophosphatidylserine in a monolayer kept at constant surface pressure (20 mN/m). Two models of adsorption of chlorpromazine in phosphatidylserine monolayers were compared. The first model correlated the amount of inserted drug molecules with the induced increase in area. The second model introduced the effect of drug adsorption on the lipid's own area by comparing the effect of increasing temperature on the lipid's own interfacial area. From the second model, the extrapolated work of insertion of one drug molecule per lipid molecule in a monolayer kept at 20 mN/m was correlated to the partition of the drug in liposomes. The work of insertion of chlorpromazine was insignificant in the unsaturated dioleoylphosphatidylserine and was maximum in the saturated distearoylphosphatidylserine monolayers. The presence of one double bond in the acyl chains dramatically reduces the work of insertion of chlorpromazine between lipid molecules and also reduces the effect chlorpromazine induces on the lipids own interfacial area in monolayers. PMID- 11403883 TI - Theoretical analysis of the molecular determinants responsible for the K(+) channel blocking by aminopyridines. AB - This work presents a theoretical analysis of the molecular determinants responsible for the pharmacological activity (K(+) channel blocking) of aminopyridines. Thus, DFT theory at the B3LYP/cc-pVDZ level is applied to a series of active compounds: 2-aminopyridine, 3-aminopyridine, 4-aminopyridine, 3,4-diaminopyridine, and 4-aminoquinoleine. The two forms present in the biological environment, neutral and cationic (protonated), are considered in vacuum as well as in aqueous solution. The results show pyramidal and planar structures for the neutral and cationic forms, respectively. An analysis of the topology of the electron density show that an increase in conjugation between the pyridine ring and the amine group is responsible for the observed planarity of the protonated forms. By computing the Laplacian of the charge density we found the pyridine nitrogen to be the preferred protonation site, as a consequence of a much higher curvature of the charge density field. Also, from three-dimensional (3D) isoLaplacian diagrams a common reactivity pattern is only found in the charged forms. This reactivity pattern implies that interaction with the biological receptor site is mediated by electrostatic interactions and hydrogen bonding. Development of a physical-mathematical model allows identification of the specific relationship of the pharmacological activity index with the affinity for the receptor and the protonation ability. PMID- 11403884 TI - Effects of intermediates on aggregation of native bovine serum albumin. AB - Protein aggregation has been recognized to be a pathological indicator for several fatal diseases, such as Alzheimer's disease, transmissible spongiform encephalopathies, Creutzfeldt-Jacob disease, etc. Aggregation usually involves conformational changes of proteins that have acquired an intermediate beta structure-rich conformation and can occur even at low protein concentration. Recent work in our laboratory has shown that bovine serum albumin (BSA), even at low-concentration, exhibits self-association properties related to conformational changes, so providing a very convenient model system to study this class of problems. Here we report data (obtained by different experimental techniques) on a mixture of BSA in native and intermediate (beta-structure-rich) form. Results show that the interaction between the two species is responsible for a decrease in the thermodynamic stability of the solution. This occurs without requiring noticeable conformational changes of the native protein. Results presented here can provide new insight on the "protein only" hypothesis proposed for the formation of plaques involved in several neurodegenerative diseases. PMID- 11403885 TI - Important inter-residue contacts for enhancing the thermal stability of thermophilic proteins. AB - Proteins from thermophilic organisms exhibit high thermal stability, but have structures that are very similar to their mesophilic homologues. In order to gain insight into the basis of thermostability, we have analyzed the medium- and long range contacts in mesophilic and thermophilic proteins of 16 different families. We found that the thermophiles prefer to have contacts between residues with hydrogen-bond-forming capability. Apart from hydrophobic contacts, more contacts are observed between polar and non-polar residues in thermophiles than mesophiles. Residue-wise analysis showed that Tyr has good contacts with several other residues, and Cys has considerably higher long-range contacts in thermophiles compared with mesophiles. Furthermore, the residues occurring in the range of 31-34 residues apart in the sequence contribute significant long-range contacts to the stability of thermophilic proteins. PMID- 11403886 TI - Theoretical formalism for bead movement powered by single two-headed motors in a motility assay. AB - Kinesins and dyneins are protein motors that can use the free energy of ATP hydrolysis to carry a cargo and move uni-directionally along a microtubule filament. The purpose of this paper is to derive the formalism connecting the ATP driven translocation reactions of these motors on microtubule filaments and the movement of the bead carried by the motor in a motility assay in which the bead is clamped at an arbitrary constant force. The formalism is thus useful in elucidating the load-dependent kinetic mechanism of the free-energy transduction of the motor using the mechanical data obtained from the motility assay. The formalism is also useful in assessing the effect on the measured motility data of various physical and hydrodynamic parameters of the assay, such as the size of the bead, the viscosity of the medium, the stiffness of the elastic element connecting the motor and the bead, etc. In a previous paper [Biophys. J. 67 (2000) 313] (hereafter referred to as paper I), we have derived the formalism for the case that the motor in the assay has only one head. In this paper we extend the derivation to the case that the motor is two-headed. The formalism is derived based on a simple two-state hand-over-hand model for the movement of the motor on microtubule, but can be easily extended to more complicated kinetic models. Effects of various hydrodynamic parameters on the velocity of the bead are studied with numerical calculations of the model. The difference between the formalism presented in this paper and the widely used "chemical" formalism, in which the movement of the kinesin and the bead is described by pure chemical reactions, is discussed. PMID- 11403887 TI - Denervation alters protein-lipid interactions in membrane fractions from electrocytes of Electrophorus electricus (L.). AB - Protein-lipid interactions are studied in normal and denervated electrocytes from Electrophorus electricus (L.). Structural modifications of the lipid micro environment encircling integral membrane proteins in membrane fractions presenting Na(+),K(+)-ATPase activity are investigated using ESR spectroscopy of stearic acid spin labeled at the 14th carbon (14-SASL). The microsomal fraction derived from the innervated electric organ exhibits, on a discontinuous sucrose gradient, a bimodal distribution of the Na(+),K(+)-ATPase activity, bands a and b. Band b is almost absent in microsomes from the denervated organ, and band a', with the same density as band a has lower Na(+),K(+)-ATPase activity. Band a' presents a larger ratio of protein-interacting lipids than band a. Analysis of the lipid stoichiometry at the protein interface indicates that denervation causes at least a twofold average decrease on protein oligomerization. Physical inactivity and denervation have similar effects on protein-lipid interactions. Denervation also influences the selectivity of proteins for fatty acids. Experiments in decreasing pH conditions performed to verify the influence of stearic acid negative charge on protein interaction revealed that denervation produces loss of charge selectivity. The observed modifications on molecular interactions induced by denervation may have importance to explain modulation of enzyme activity. PMID- 11403888 TI - Aromatase expression and activity in male and female cultured rat hypothalamic neurons: effect of androgens. AB - Aromatase is possibly involved in male brain sexual differentiation. Aim of these experiments was to evaluate the role of testosterone (T) and of DHT, in the regulation of aromatase expression and activity. The experiments were done utilizing rat primary cultures of hypothalamic neurons from 16-day old embryos sex-screened by SRY gene. Aromatase expression was assessed semiquantitatively by RT-PCR using a neuronal marker (MAP2c) as coamplification product; enzymatic activity was estimated by the 3H(2)O method. The results indicate that (1) cultured neurons possess a functional aromatase, which increases significantly during a 5-days culture period; (2) neurons from males possess a higher expression and activity of the enzyme than females; (3) androgens negatively control expression/activity of aromatase in males, DHT is more active than T; (4) on the contrary, in females T produces a small stimulation of aromatase expression, but not of activity (DHT has produced inconsistent results). The results obtained in this model indicate that T does not stimulate aromatase; therefore, it is not responsible for triggering the perinatal enzymatic peak, nor for the sexual dimorphic aromatase expression. A model is proposed in which DHT might induce, at least in males, the descending phase of the aromatase peak. PMID- 11403889 TI - Aromatase expression in prepuberal Sertoli cells: effect of thyroid hormone. AB - Aromatase activity has recently been assumed as a Sertoli cell functional maturation marker since it is maximally expressed in prepuberal age then it dramatically decreases at puberty and is virtually absent in adult age. Neonatal hypothyroidism is associated with a prolonged proliferation of Sertoli cells. This immature stage persists concomitantly with a dramatic enhancement of aromatase activity reversed by triiodothyronine (T3) either in vivo or in vitro administration. Therefore, in the present study, after immunolocalisation of aromatase in the cytoplasm of cultured Sertoli cells as well as in testis section, we investigate the regulatory effects of T3 in the same cells just at the age when aromatase activity is reported to be maximally expressed. In this aim, the effects of thyroid hormone have been evaluated in 2-weeks-old rats, in basal condition and upon stimulation with dibutyryl cyclic AMP [(Bu)(2)cAMP] by simultaneously analysing three functional levels of aromatase, mRNA expression; protein content; enzymatic activity. Western-blot analysis of Sertoli cell extracts revealed a protein, which co-migrated with a 55 kDa protein detected in human placenta used a positive control. The presence of a functional P450 aromatase protein in purified Sertoli cells was confirmed by the measurement [3H]H(2)O released after incubation with [1beta-(3)H]androst-4-3,17-dione. At the dose used, T3 down-regulates basal aromatase activity, while aromatase mRNA expression was apparently not inhibited. It is noteworthy that aromatase content pattern evaluated by Western blot analysis did not tightly parallel the aromatase activity pattern which clearly displays the inhibitory effects of T3, in basal condition ad upon (Bu)(2)cAMP stimulation, simulating FSH stimulation. The detection of mRNA altered transcript coding for putative protein lacking both aromatic and heme-binding regions upon T3 treatment and unable to convert androgens into estrogens, provides a reasonable explanation for the observed discrepancies between aromatase protein pattern, P450arom mRNA levels and aromatase activity. The authors conclude that although the altered transcript induced by prolonged exposure to T3 is a mechanism by which T3 may down regulate aromatase activity, it cannot be ruled out a direct effect of this hormone at the transcription levels since a recognisable emisite for potential TR(s) binding is located in the promoter region of aromatase gene. Thus a further investigation on T3 modulator role on aromatase gene promoter should be pursued even utilising higher doses of T3. PMID- 11403890 TI - Aromatase expression in the human male. AB - The role of estrogens produced by the testis may involve negative feedback regulation of androgen biosynthesis. Estrogens are also associated with contractile processes of seminiferous tubules, and might have mitogenic effects on Sertoli and Leydig cells. To investigate the location of aromatase (estrogen synthetase) in the testes, tissue from normal human subjects, aged 3 months to 72 years were studied using immunocytochemistry. In mature testes, aromatase immunostain was always associated with Leydig cells and was absent from Sertoli cells. Aromatase activity ranged from 0.014-0.55 pmol estrogen per mg/h and was significantly correlated with the immunostain intensity (P<0.02). Activity and immunostain intensity did not correlate with increasing age. Rather, the highest levels were measured in four of six testes of men aged 18-20 years, three of whom also had the strongest immunostain in larger and more prominent Leydig cell clusters than those in the other specimens. A low level of aromatase activity but no immunostain was detected in prepubertal testes. However, in several prepubertal patients with Peutz-Jegher's Syndrome (PJS) with bilateral multifocal sex cord tumors and enlarged seminiferous tubules and Sertoli cells, aromatase was expressed in these Sertoli cells, but absent from normal Sertoli and Leydig cells. Increased aromatase expression in these tissues involved activation of upstream regulatory elements of the gonadal P II promoter of P-450(arom). In a prepubertal boy with gynecomastia but without PJS, aromatase excess appeared to be due to increased aromatization in skin fibroblasts and lymphocytes. Several members of the patient's family including his sister also expressed high levels of aromatase. This condition appears to be inherited in an autosomal dominant manner. PMID- 11403891 TI - Oestrogen, its receptors and function in the male reproductive tract - a review. AB - Oestrogen is synthesized in the male reproductive system by at least three different cell types; Sertoli, Leydig and germ cells. Although testosterone is recognized as the primary sex steroid in man, oestrogen is produced in sizable quantities in the testis, as well as the brain and is found in extremely high concentrations in the semen of several species. The high concentration of oestrogen in rete testis fluid of the rodent is now thought to be derived from the conversion of testosterone to estradiol by P450 aromatase in germ cells of the testis and spermatozoa traversing the reproductive tract. This new major source of oestrogen would target oestrogen receptors in the male reproductive tract, in particular the efferent ductules, which contain the highest concentration of oestrogen receptor-alpha. This recent data raises new hypotheses regarding the role of oestrogen in the function of the male reproductive system. The oestrogen receptor-alpha knockout mouse was used to help define the function of oestrogen in the male. It was found that oestrogen receptor-alpha is essential for fluid reabsorption in the efferent ductules and in the absence of expression the male is infertile. PMID- 11403892 TI - Effects of estrogenic compounds on human spermatozoa: evidence for interaction with a nongenomic receptor for estrogen on human sperm membrane. AB - Estrogens play an important role in the development and regulation of the male reproductive system. We have earlier shown that a nongenomic receptor for estradiol present on sperm plasma membrane mediates the effects exerted by this hormone on sperm intracellular calcium concentrations ([Ca(2+)](i)), as well as on the biological response to progesterone (P). In particular, 17 beta-estradiol (17betaE(2)) shows an inhibitory effect on P-mediated calcium influx and acrosome reaction (AR). In the present study, the effects of different anti-estrogens and xenoestrogens on [Ca(2+)](i) and AR stimulated by P have been investigated in human spermatozoa in order to better define the pharmacological characteristics of the sperm membrane estrogen receptor. The anti-estrogens tamoxifen (Tx) and ICI 164384 (ICI) induce only a slight increase of [Ca(2+)](i), which, however, as in the case of 17betaE(2), results in a reduction of P-stimulated calcium influx. Moreover, both the compounds reduce the calcium response to 17betaE(2) without affecting 17betaE(2)-inhibition of calcium response to P. Concerning AR, Tx alone does not alter either spontaneous or P-stimulated AR but partially revert the inhibitory effect of 17betaE(2). These results indicate that the two estrogens act as pharmacological agonists of the membrane estrogen receptors of human spermatozoa. On the other hand, the xenoestrogens bisphenol A (BPA) and octyiphenol polyethoxilate (OP) do not exert any direct effect on calcium fluxes and AR in human spermatozoa either in basal conditions or in response to P challenge. Moreover, although these environmental estrogens have been suggested to mimic estrogen effects in the other cell types, probably acting through genomic receptors, in human spermatozoa they do not interfere with 17betaE(2) binding to its membrane receptor and with the short-term effects exerted by this steroid. In conclusion, our data indicate that the membrane receptor for estradiol in human spermatozoa shows both biochemical and pharmacological differences respect to the genomic receptor. PMID- 11403893 TI - Estrogen receptor beta expression in human prostate tissue. AB - Estrogen receptor subtype beta (ERbeta) is highly expressed in rat prostate epithelium, but its presence in human prostate needs to be confirmed. Here we investigated the expression of ERbeta in five benign (normal and/or hyperplastic) and 10 malignant (Gleasons' score 2-7) prostate tissue specimens using immunohistochemistry. Immunohistochemistry was performed on formalin-fixed, paraffin-embedded tissue sections, using a commercially available ERbeta polyclonal antibody developed against the C-terminal amino acid residue. Nuclear ERbeta expression was found in the nuclei of glandular epithelium of benign prostate tissue specimens; faint nuclear ERbeta positivity was also present in a few stromal cells around normal epithelium. Nuclear ERbeta specific immunostaining was undetectable in all prostate cancer sections. PMID- 11403894 TI - Estrogens and health in males. AB - Estrogen receptor concentrations are higher in the male reproductive tract than in other organs. Brain structure, neuronal organization and behavioral sex differences result from brain conversion of testosterone into estradiol within the brain. Estrogens modulate hormonal secretion at pituitary level and immune function at thymus level. Estrogens promote vasodilatatory and protective effects on the cardiovascular system by acting on the vascular smooth muscle and endothelium. Adult men with mutations in genes for estrogen receptor or aromatase are affected by osteopenia and tall stature, open epiphysis, which is corrected by estrogen treatment. Over the past few years there has seen a decline in sperm concentration, which has been attributed to exposure of fetal testes to estrogens. Many substances have estrogen-like properties and inhibit the action of estradiol or testosterone action. In conclusion, estrogens play a pivotal role in men also. In particular, taking in account their prevalent origin from testosterone aromatization at tissue and peripheral levels the presence and the distribution of the two receptors (ERalpha and ERbeta) are responsible for different responses in physiological and pathological conditions. PMID- 11403895 TI - Estrogen receptor-alpha is required by the supporting somatic cells for spermatogenesis. AB - The gene for estrogen receptor-alpha (ERalpha) was disrupted in embryonic stem cells by homologous recombination and these cells were used to generate mice with a targeted mutation in the ERalpha gene (alphaERKO mice). It was found that males homozygous for the mutation are infertile, indicating that estrogen signaling through this nuclear hormone receptor is required for male reproductive function. Although spermatogenesis appears normal in juvenile and young adult alphaERKO mice, the sperm produced are unable to fertilize eggs in vitro. To determine whether ERalpha is required by somatic or germ cells in the male reproductive tract, we transplanted germ cells from homozygous mutant (ERalpha(-/-)) males to the testes of wild-type (ERalpha(+/+)) males depleted of germ cells by busulfan treatment. The recipients ('surrogate fathers') sired offspring heterozygous for the mutation (ERalpha(+/-)) and carrying the coat-color marker of the infertile donor males. This indicated that ERalpha(-/-) germ cells are able to produce sperm competent to fertilize when they are supported by ERalpha(+/+) somatic cells. When ERalpha(+/-) offspring produced by germ cell transplantation were mated to produce ERalpha(-/-) males, these mice were found to have the same phenotype as originally reported for alphaERKO males. These studies showed that male germ cells do not require ERalpha for regulation of their own genes for development and function, and strongly imply that somatic cells of the male reproductive tract require ERalpha to support the production of sperm that are capable of fertilization. PMID- 11403896 TI - Germ cells: a new source of estrogens in the male gonad. AB - The cytochrome P450 aromatase (P450arom) is a key enzyme responsible for the formation of estrogens from androgens and is present in the endoplasmic reticulum of various tissues. P450arom has been immunolocalized in Leydig cells of numerous species as well as in germ cells of mouse, bank vole and brown bear. Aromatase activity has been measured in vitro in immature and mature rat Leydig cells and Sertoli cells, whereas in pig, ram and humans the enzyme activity is only present in Leydig cells. In the mature rat testis we have used complementary approaches to demonstrate that not only somatic cells but also germ cells represent a new source of estrogens. In pachytene spermatocytes and Leydig cells, the amount of P450arom mRNA measured by a quantitative competitive RT-PCR method is 10-fold higher than in Sertoli cells. According to the stage of the germ cell maturation, the amount of aromatase transcripts decreases, being more elevated in younger than in mature rat germ cells. By contrast, the aromatase activity in the microsomal fractions is two- to four-fold greater in spermatozoa when compared to the two other enriched germ cell preparations used. We have immunolocalized the P450arom in elongated spermatids and spermatozoa. Moreover, we described the existence of alternative splicing events of P450arom mRNA in pachytene spermatocytes and round spermatids that are not likely to encode functional aromatase molecules. Therefore, the aromatase gene expression and its transduction in a fully active protein in rat germ cells evidences an additional site for estrogen production within the testis of some mammals. Taking into account the large distribution of estrogen receptors in the testicular cells, we begin to understand the physiological role of these female hormones in the male gonad. PMID- 11403897 TI - Modulation of gene expression by androgen and oestrogens in the testis and prostate of the adult rat following androgen withdrawal. AB - Androgens are important for the structural and functional integrity of the testis and the prostate and this may in part be mediated by the aromatisation of testosterone to oestradiol. The aim of the present study was to establish an in vivo model that would allow the identification of genes, the expression of which was regulated acutely by androgen and/or oestrogen in the male reproductive system. In rats in which the Leydig cells were ablated by administration of ethane dimethane sulfonate (EDS) 6 days earlier, testosterone esters (T) were administered from day 0 (To), and additional animals were administered either T, 17beta-oestradiol benzoate (EB) or diethylstilbestrol (DES) for 1 or 4 h on day 6 after EDS-treatment. Nuclear immunoexpression of the androgen receptor (AR) was reduced or absent from the testis but unaffected in the ventral prostate following these treatments. ERbeta immunoexpression in these tissues was unchanged. Northern blot analysis showed that EB and DES as well as T administration 4 h earlier could modulate mRNA expression of two androgen responsive genes, C3 and SGP-2, in the prostate. The co-administration of T or EB with the AR antagonist, flutamide, or with the ER antagonist, ICI 182,780 (ICI), did not block the suppression of SGP-2 mRNA expression by T or EB. In contrast, the upregulation of C3 mRNA expression by T was successfully antagonised by both flutamide and by ICI. A preliminary evaluation of the expression of three Sertoli cell and five germ cell mRNAs revealed that their expression was not steroid regulated. Our results support the hypothesis that the action of testosterone in the male reproductive system may in part be mediated by its conversion to oestradiol. This in vivo model should prove of value in future studies to identify androgen and oestrogen regulated genes in the male reproductive system. PMID- 11403898 TI - Estradiol enhances the stimulatory effect of FSH on testicular maturation and contributes to precocious initiation of spermatogenesis. AB - Male rats were daily injected with human FSH (hFSH) or estradiol benzoate (EB) or hFSH+EB between day 5 and 15 of life and autopsied on day 16. hFSH accelerated testicular growth, increased number of spermatogonia and serum level of testosterone. hFSH stimulated also spermatogonia differentiation, which resulted in 5-fold increase of the number of spermatocytes. EB given alone induced adverse, inhibitory effects on spermatogenesis and serum testosterone, did not influence serum FSH and LH but increased 14-fold the level of prolactin. Except from testosterone, EB given with hFSH not only overcame inhibitions, but multiplied hFSH stimulatory effects on spermatogenesis up to 30-times of control values. In addition, after FSH+EB premeiotic germ cell ratio reached adult type value precociously. Estradiol may play regulatory roles in testicular maturation (1) inhibitory, direct one or resulting from decrease in testosterone secretion; (2) stimulatory, by enhancement of FSH action, with a possible involvement of prolactin that may act in concert with FSH. PMID- 11403899 TI - Environmental xenoestrogens, antiandrogens and disorders of male sexual differentiation. AB - Over the past 20 years, the documented increase in the disorders of male sexual differentiation, such as hypospadias, cryptorchidism, and micropenis, has led to the suspicion that environmental chemicals are detrimental to normal male genital development in utero. Male sexual differentiation is critically dependent on the normal action of androgens, and unbalanced androgen/estrogen ratios can disturb it. Environmental xenoestrogens (such as herbicides, pesticides, PCBs, plasticizers, and polystyrenes) that mimic estrogens or environmental antiandrogens (such as polyaromatic hydrocarbons, linuron, vinclozolin, and pp'DDE) that disturb endocrine balance, cause demasculinizing effects in the male foetus. These environmental chemicals are often referred to as endocrine disruptors: they are thought to mimic endogenous estrogens by entering the cell, binding to the receptor and activating transcription, they may also antagonize normal androgen action. We have established numerous cell lines to assess the estrogenicity and antiandrogenicity of compounds found in the environment and to identify new products present in wastewater effluents that are able to disrupt endocrine functions. Several cell lines responding to estrogens have been obtained in our group, including cells with different enzymatic equipment and cells expressing chimeric receptor or natural estrogen receptors alpha and beta. These cell lines have proved to be useful for assessing the biological activity of pesticides, fungicides, and chemicals found in plastic or discarded in the environment. In order to generate a powerful tool for the investigation of androgen action and the rapid screening of potential antagonists, we developed a new stable prostatic cell line. The PALM cell line is an original cellular model to characterize the response of hAR, and it provides an easy and rapid bioluminescent test to identify new antagonists. We also developed a model based on a fusion protein between the androgen receptor (AR) and the green fluorescent protein (GFP) to study the intracellular dynamics of AR. The GFP-AR model was applied to define the ability of several xenoestrogens and antiandrogens to inhibit the nuclear transfer of AR. The ubiquitous presence of endocrine disruptors in the environment and the increased incidence of neonatal genital malformation support the hypothesis that disturbed male sexual differentiation may in some cases be caused by increased exposure to environmental xenoestrogens and/or antiandrogens. PMID- 11403900 TI - Congenital estrogen deficiency: in search of the estrogen role in human male reproduction. AB - Recently, a remarkable progress has been made in our understanding about the role of sex steroids in male physiology. In this paper, we consider the clinical aspects of congenital estrogen deficiency - notably, estrogen resistance and aromatase deficiency - in men and we discuss both well-established and supposed estrogen roles in the human male reproductive function. These topics include the role of estrogens in the control of gonadotropin secretion, in male fertility determination and psychosexual behavior. Briefly, estrogens play a pivotal role in the control of serum gonadotropin concentrations in the human male. Furthermore, a possible role of estrogens on both human male fertility and sexuality has also been suggested by recent studies, even though the available data are far from being conclusive. Conversely, for what concern fertility and sexual behavior, a well-established effect of estrogens has been provided by recent studies on male rodents, which show impaired sexual behavior and fertility as a consequence of estrogen defect. PMID- 11403901 TI - Aromatase inhibitors: past, present and future. AB - For the cellular physiology of sex steroid sensitive cells, the androgen/estrogen ratio may be more important than only one hormone action per se, in both sexes. This ratio is controlled in vertebrates by aromatase; its gene expression can be inhibited in different ways, and this is crucial for the treatment of estrogen dependent diseases such as breast cancer, or gynecomastia in males for instance. To reach this goal, new steroidal and non-steroidal inhibitors are continuously being developed, and some of them are used as first or second line agents. Aromatase inhibition is also an essential tool for studying the role of estrogens in the adult, or during development. Aromatase inhibitors have shown in particular that estrogens are essential also in males for skeletal maturation and bone mineralization, development of masculine dendritic morphology in male brain linked to mating behaviour, and testicular function. Testosterone is often the prohormone converted in situ in active estrogens, at these levels. Several strategies can be used for aromatase inhibition. The first ones employed were blind screening or deductions from in vivo observations, which led for instance to the discovery of the role of aminoglutethimide in aromatase inhibition. Subsequently, in the years 1975-1990, the molecular modeling of compounds to mimic the substrate shape of the enzyme constituted the major idea. Hundreds of chemicals were synthesized by numerous authors, ranging from the well-known and very efficient 4-OHA to complicated imidazole or indane derivatives tested by sophisticated comparative molecular field analyses. Reticulum-bound active aromatase has not as yet been X-ray analyzed. Thus, aromatase inhibitors were also used more recently to probe and understand the active site conformation of the enzyme and its modelization was obtained from comparisons with bacterial related cytochromes. We developed a mammalian model considerably closer to human aromatase in order to study the active site shape with new potent aromatase non steroidal inhibitors. This model is equine aromatase. This enzyme was biochemically characterized, purified, and cloned by our group. It allowed testing, by site-directed mutagenesis, predictive hypotheses in human aromatase which contributed to designing of new inhibitors. The understanding of the functioning of an essential member of the cytochrome P450 family, which is necessary for cellular detoxification, was also facilitated. Inhibition of aromatase activity has also been carried out with antibodies directed to the catalytic site and at the gene level by knock-out or by control of factor specific promoters. This may result in different mRNA synthesized by alternative splicing. We have also obtained specific inhibition of aromatase activity in human cells with antisense stable phosphorothioate oligodeoxynucleotides directed against aromatase mRNA tertiary structures. Besides known steroidal and non steroidal inhibitors, the antiaromatase effects of compounds found in our daily environment such as dietary flavonoids or xenobiotic pollutants have also been described. Finally, we underline that all these aromatase inhibitors, or methods of aromatase inhibition, can modulate the estrogenic balance essential not only for female, but also for male physiology, including gonadal function. PMID- 11403902 TI - Aromatase gene expression in the stallion. AB - Adult stallion secretes very high estrogen levels in its testicular vein and semen, and the responsible enzyme cytochrome P450 aromatase (P450 arom) is known to be present mainly in Leydig cells. We studied in further details the distribution of equine aromatase in various adult tissues including the brain (hypothalamic area), liver, kidney, small intestine, muscle, bulbourethral gland and testes. The aromatase mRNA was essentially detected by RT-PCR in testis (169+/-14 amol of aromatase mRNA per microg of total RNA) and was barely detectable in brain, or below 0.1 amol/microg RNA in other tissues. This range of expression was confirmed by ELISA (50+/-7 pg/microg total protein) in the testis, and by immunoblot, evidencing a 53 kDA specific protein band in testis and brain only. The corresponding aromatase activity was well detected, by 3H(2)O release from 1beta, 2beta(3)H-androstenedione, in testis and brain (200+/-23 and 25+/-6 pmol/min per mg, respectively) and below 3 pmol product formed/min per mg in other tissues. This study indicates that the testis, among the tissues analyzed, is the major source of aromatase in the adult stallion, and that the aromatase gene expression is specifically enhanced at this level, and is responsible for the high estrogen synthesis observed. Moreover, the study of aromatase in one colt testis has shown lower levels of transcripts, protein and enzyme activity, evidencing that aromatase is regulated during the development and may serve as a useful marker of testicular function. As the second organ where aromatase mRNA and activity are both well detected is brain, this study also underlines the possible role of neurosteroids in stallion on behaviour, brain function or central endocrine control. PMID- 11403903 TI - Regulation of the aromatase gene expression in mature rat Leydig cells. AB - Besides gonadotrophins and testosterone, numerous intratesticular factors, and among them estrogens play a crucial role in the regulation of spermatogenesis in the mammalian testis. The ability of the male gonad to convert androgens into estrogens is well known; the microsomal enzymatic complex involved in this transformation named aromatase, is composed of a specific cytochrome P450 aromatase (P450arom) and a ubiquitous reductase. Using a highly specific reverse transcription polymerase chain reaction (RT-PCR) method, we have measured the amount of P450arom mRNA in purified rat Leydig cells submitted to different treatments during 24 h. In parallel, the estradiol output was determined by RIA. Whatever the concentrations of testosterone used (as substrate of aromatase activity), we noted an increase of the estradiol production, the maximum being obtained for 200 ng/ml (28%). Related to the P450arom mRNA levels, a decrease was first observed for 50 and 100 ng/ml of testosterone then an increase (20%) for the higher doses (200-500 ng/ml). The addition of oLH (0.1-50 ng/ml) to the Leydig cells culture medium induced a dose-related increase of estradiol output till 10 ng/ml. For 50 ng/ml, a decrease was observed. Dealing with the mRNA levels, we first recorded a diminution for 0.1-1 ng/ml of oLH, which was abolished by the addition of testosterone. Then the mRNA levels were increased and reached a maximum for 5-10 ng/ml of oLH (35 and 50%, respectively, in absence and in presence of testosterone). The addition of oLH (50 ng/ml) induced a large augmentation of the quantity of P450arom mRNA (1.9 and 2.1-fold, respectively, in absence or in presence of testosterone). DbcAMP mimicked the effects of oLH. From these data, we confirm the presence of cAMP response-like elements (CRE) and the existence of androgen responsive elements (ARE) sites on the P450arom gene in rat Leydig cells. PMID- 11403904 TI - The role of estrogen and estrogen receptor-alpha in male adipose tissue. AB - Males and females both express estrogen receptor (ER) in white adipose tissue (WAT), and estrogens appear to play an important role in regulating WAT in females. However, the role of ER in male WAT was unclear. In this review, we describe our work, which used wild type (WT) and ERalpha-knockout (alphaERKO) male and female mice to determine the role of ERalpha in regulating WAT and brown adipose tissue (BAT). There were progressive increases in WAT with advancing age in alphaERKO compared with WT males; weights of various WAT depots in alphaERKO males were increased by more than 100% compared with WT controls during adulthood. Conversely, BAT weight was similar in alphaERKO and WT males at all ages. Adipocyte areas and numbers were also increased in WAT from alphaERKO compared with WT males. Compared with WT controls, alphaERKO females also had increases in WAT. The alphaERKO mice also had insulin resistance and impaired glucose tolerance, similar to humans lacking ERalpha or aromatase. The obesity in alphaERKO males appeared to involve decreased energy expenditure rather than hyperphagia. In summary, ERalpha absence causes adipocyte hyperplasia and hypertrophy in WAT, but not BAT, and is accompanied by insulin resistance and glucose intolerance in both males and females. These results are the first evidence that the estrogen/ERalpha signaling system is critical in female and male WAT deposition, and may have clinical implications. PMID- 11403905 TI - Tissue-specific expression of human ERalpha and ERbeta in the male. AB - The important role of estrogens in women in physiological and pathological processes is well accepted, but recently it has become evident that estrogens are also important in male physiology, in particular, within bone metabolism and reproduction. Consequently, it is necessary to identify and to characterize the molecular mechanisms of estrogen action in order to evaluate how the pleiotropic effects of estrogens are mediated in a variety of tissues. We have recently shown that human estrogen receptor alpha (ERalpha) mRNA is transcribed from at least six different promoters (1A-1F). Transcription of ERalpha in bone is exclusively dependent on the F-promoter. To study the regulation of ER expression in this tissue, we examined 1 kbp of the F-promoter region of human ERalpha, which is located more than 70 kbp upstream of the transcription start site of the ERalpha gene. Transient transfection experiments demonstrated a basal activity from the F promoter, which was further increased when ERalpha was cotransfected. We have shown recently that the F-promoter can give rise to at least two ERalpha isoforms in bone. On the contrary, ERbeta expression in primary osteoblasts is extremely low, indicating that this ER isoform plays only a minor role in these cells. In contrast to bone, we have demonstrated that both ERalpha and ERbeta transcripts are readily detected in testis. Here, we report that besides ERalpha, ERbeta transcripts can give rise to two protein isoforms and that this complex situation could have important functional consequences for the signalling of estrogens and their analogs. PMID- 11403906 TI - beta-estradiol 3-benzoate affects spermatogenesis in the adult mouse. AB - beta-estradiol 3-benzoate (E(2)B) (10, 16, 20, 40, 80 and 160 microg/kg body weight) was administered daily to experimental groups of adult mice for the following periods; 2, 3 days, 1, 2, 4, and 8 weeks. Morphological changes in the testes were observed by both light and electron microscopy. Exfoliation of the germ cells was observed in the lumen of the seminiferous tubule. The spermatogenic cycle, especially stage XII, was disordered. Spermatids older than step 6 were severely affected. Detected abnormalities in the spermatids were deformation of the nucleus and acrosome. Partial deletion in the Sertoli spermatid ectoplasmic specialization was also observed. Germ cells younger than step 7 spermatids were not affected morphologically. These abnormalities were not detected in the mice treated with the chemical at less than 16 microg/kg body weight. It is concluded that the chemical seems to affect round spermatids metabolically, but the morphological effect can be detected only from the spermatids older than step 6. The effects of the chemical on adult mice were reversible. PMID- 11403907 TI - Prenatal testosterone and luteinizing hormone levels in male rats exposed during pregnancy to 2,3,7,8-tetrachlorodibenzo-p-dioxin and diethylstilbestrol. AB - Changes in the perinatal testosterone surge have been related to demasculinization of the central nervous system and androgen-dependent growth of the reproductive organs in male mammals. Earlier reports suggest that 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) interferes with androgen production, but the perinatal effects have remained elusive. In the present study we explored in utero-effects of TCDD (0.05, 0.1, 0.5, and 1.0 microg/kg), introduced on day 13.5 of pregnancy, on prenatal (day 19.5 post-conception [p.c.]) testosterone (T) surge and pituitary luteinizing hormone (LH) production in TCDD-resistant Han/Wistar (H/W) and TCDD-sensitive Long-Evans (L-E) rats. To elucidate estrogenic effects on T and LH production, Sprague-Dawley (S-D) fetuses with previously known DES-sensitivity were exposed in utero to diethylstilbestrol (DES, 100-300 microg/kg) on days 13.5, 15.5, and 17.5 p.c. For comparison, H/W fetuses that responded to TCDD treatments were exposed to DES at concentration of 100 microg/kg. It was found that TCDD has a stimulatory effect on testicular T synthesis in the H/W fetuses and that their circulating T concentrations increased significantly. The effect was not seen in the inbred L-E fetuses, which throughout the study showed considerably low testicular T levels. Pituitary LH concentrations also increased in the H/W fetuses exposed to TCDD. Effects of TCDD (1.0 microg/kg) in the H/W fetuses could be confirmed in vitro by human chorionic gonadotropin (hCG) stimulation assay showing the highest response rate in the TCDD exposed testes. Stimulation of cyclic AMP (adenosine-3', 5'-cyclic monophosphate[cAMP]) production was not considerably altered by in utero TCDD exposure. A significant depression in testicular and plasma T content was seen in the DES-exposed S-D and H/W fetuses, but pituitary LH levels did not alter considerably. In the presence of hCG, DES-exposed testes showed lower in vitro T and cAMP production rates compared to the untreated testes. TCDD (1.0 microg/kg) increased and DES decreased the male body weight gain, but the changes were not sex-dependent. It is concluded that TCDD may increase the amplitude of the prenatal testosterone surge in male rats by stimulating pituitary LH production and enhancing the sensitivity of the fetal testis to LH. DES, on the contrary, apparently impairs testicular steroidogenesis and pituitary function. PMID- 11403908 TI - Extracellular ATP stimulates estradiol secretion in rat Sertoli cells in vitro: modulation by external sodium. AB - In this study, we examined the effects of extracellular ATP (ATPe) on [Ca(2+)](i), [Na(+)](i), plasma membrane potential changes and estradiol secretion in rat Sertoli cells. ATPe caused a rapid rise of [Ca(2+)](i) with an initial spike followed by a long lasting plateau. The first rapid spike was dependent on the release of Ca(2+) from internal stores as it also occurred in Ca(2+)-free medium while the long lasting plateau phase was dependent on Ca(2+) influx from the external medium. ATPe stimulated a rapid plasma membrane depolarization that was dependent on an influx of Na(+) from the external medium as demonstrated by plasma membrane potential monitoring in Na(+)-free medium and by [Na(+)](i) measurement with the Na(+)-sensitive fluorescent dye SBFI. ATPe stimulated estradiol secretion in a dose dependent manner and was fully dependent on the presence of Na(+) in the external medium while the presence of Ca(2+) was not necessary. Among the different nucleotides tested, only ATP, ATP-5'-[gamma thio]triphosphate, UTP, alpha,beta-methylene-ATP were effective in stimulating estradiol secretion. These results demonstrate that rat Sertoli cells possess P2 purinergic receptors belonging to the P2X and P2Y subfamily which activation induces [Ca(2+)](i) and [Na(+)](i) rise and Na(+)-dependent plasma membrane depolarization leading to estradiol secretion. PMID- 11403909 TI - Photoperiod-dependent capability of androgen aromatization and the role of estrogens in the bank vole testis visualized by means of immunohistochemistry. AB - Detection of steroid hormone receptors within a target tissue is important for an understanding of their crucial role in regulating of steroids' action. In the light of recent knowledge on the role of estrogens in male gonads the efforts were undertaken to clarify and discuss a role of androgen receptors, aromatase and estrogen receptors (ER) in mediating testosterone and/or estradiol action in testicular cells of bank voles that were kept under short or long light cycles. Immunohistochemistry was performed on paraplast embedded sections of the bank vole testes. First, androgen receptors were immunolocalized in testicular somatic cells while germ cell did not express any immunoreaction. Moreover, the ability to convert androgens to estrogens by various testicular cells was documented; aromatase immunoexpression was found in testis sections, not only in Leydig cells and Sertoli cells but also in germ cells. Finally, the expression of estrogen receptor-alpha (ERalpha) was observed in Leydig cells whereas the presence of estrogen receptor-beta (ERbeta) was detected in Sertoli and germ cells, namely spermatocytes and spermatids. The cellular distribution of androgen receptors appeared to be light -and age-dependent in adults; immunoexpression of aromatase and ERbeta was found to be both age -and photoperiod-dependent in germ cells. PMID- 11403910 TI - Effect of tamoxifen on sperm fertilising ability and preimplantation embryo development. AB - Recent evidences point to a role of estrogens in males. We have earlier reported that tamoxifen, a synthetic non-steroidal antiestrogen, when administered to adult male rats, in the dose range of 0.04-0.4 mg/kg per day, reduced fertility. The reduced fertility was measured in terms of fertility index (a measure of the efficiency of the ovulated ovas to fertilise and implant), fecundity (siring ability) and litter size. The present study was done to investigate whether the reduction in fertility index was due to reduction in fertilising ability or increase in pre-implantation embryo loss. Also a dose related effect of tamoxifen from 0.02 mg to 2 mg/kg per day on the fertility of the male rats was studied. To study the fertilising ability, control and tamoxifen (0.4 mg/kg per day, the most effective dose) treated adult male rats were mated with normal cycling females and the females sacrificed at day 0-4 of gestation. Eggs fertilised/unfertilised were flushed from the oviduct/uterus and the number and types of eggs were noted. The index of fertilisation, a measure of the fertilising ability was determined. The studies demonstrate that the reduction in fertility is not due to decreased fertilising ability but because of the increased pre-implantation embryo loss as evident from an increase in number of abnormal eggs in the treated group with no change in index of fertilisation. A dose related decrease in fertility was observed. The present study suggests that tamoxifen at 0.02-2-mg dose is predominantly estrogenic in males and paternal factor/s sensitive to tamoxifen is involved in embryogenesis. PMID- 11403911 TI - Induction of hepatic aromatase by p,p'-DDE in adult male rats. AB - 1,1-dichloro-2,2-bis(p-chlorophenyl)ethylene (p,p'-DDE, DDE) is a stable metabolite of the pesticide DDT and a persistent environmental pollutant. Earlier reports have demonstrated that DDE is an endocrine-active compound capable of affecting early-stage sexual differentiation in male rats. Experiments based on receptor binding affinity and receptor-mediated transcriptional activation have identified DDE as an androgen receptor antagonist. Other effects of DDE include modulation of the expression and activity of cytochrome P450 (CYP) enzymes, some of which function as steroid hydroxylases, and elevation of serum estrogen levels in treated male rats. These effects suggest the possibility of DDE-caused induction of aromatase, a member of CYP family that catalyzes the conversion of C19 steroids to estrogens. The present study was conducted to determine whether hepatic aromatase was responsive to DDE treatment. We found that hepatic aromatase protein in adult male rats was greatly increased after seven daily oral treatments of DDE at a dosage of 100 mg/kg wt. per day. This induction was seen in both immunoblot and immunohistochemistry of liver tissue sections. Distribution of the aromatase in the liver corresponded to the distribution of hypertrophic hepatocytes in the tissue. Furthermore, we found a large increase in hepatic microsomal aromatase activity in DDE-treated animals, although the difference in serum 17beta-estradiol concentrations between treated animals and controls was not statistically significant. However, an in vitro experiment using primary culture of rat hepatocytes did not show a change in aromatase level after DDE treatment at four concentrations ranging from 0 to 5x10(-6) M for 24 h. Meanwhile, CYP 2B1 induction, a known DDE effect in primary rat hepatocyte culture, was seen in those cells. This study supports the notion that induction of aromatase by DDE is a contributory factor to its reproductive developmental effects. PMID- 11403912 TI - Role of estrogen on bone in the human male: insights from the natural models of congenital estrogen deficiency. AB - The reports of congenital estrogen deficiency - notably, estrogen resistance and aromatase deficiency - have completely changed our knowledge on the role of estrogen on bone in males. Particularly, the bone changes at puberty, which were classically considered androgen-dependent, are now considered to be induced at least in part by estrogen action. Clinical cases of congenital estrogen deficiency have clearly demonstrated that the role of estrogens in epiphyseal closure, skeletal proportions and bone mineralization is crucial not only in women but also in men. In addition progress have been made in the treatment of such a rare disease even though further studies are needed to a definitive understanding of this issue. PMID- 11403914 TI - Telomeres and replicative senescence: Is it only length that counts? AB - Telomeres are well established as a major 'replicometer', counting the population doublings in primary human cell cultures and ultimately triggering replicative senescence. However, neither is the pace of this biological clock inert, nor is there a fixed threshold telomere length acting as the universal trigger of replicative senescence. The available data suggest that opening of the telomeric loop and unscheduled exposure of the single-stranded G-rich telomeric overhang might act like a semaphore to signal senescent cell cycle arrest. Short telomere length, telomeric single-strand breaks, low levels of loop-stabilizing proteins, or other factors may trigger this opening of the loop. Thus, both telomere shortening and the ultimate signalling into senescence are able to integrate different environmental and genetic factors, especially oxidative stress-mediated damage, which might otherwise become a thread to genomic stability. PMID- 11403915 TI - Attenuation by ambroxol of monochloramine-enhanced gastric carcinogenesis: a possible prevention against Helicobacter pylori-associated gastric carcinogenesis. AB - The effects of combined administration of a reactive oxidant, monochloramine, and a mucoregulatory agent, ambroxol, on the development of gastric cancers induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) were investigated in inbred Wistar rats. After receiving oral MNNG and regular chow pellets for 25 weeks, rats received regular chow pellets or chow pellets containing 20% ammonium acetate, and normal tap water or water containing 30 mM sodium hypochlorite, with or without subcutaneous injection of ambroxol at high or low doses, until the end of the experiment at week 52. Treatment with both ammonium acetate and sodium hypochlorite, which produce monochloramine, significantly increased the incidence of gastric cancers at week 52, whereas concomitant administration of ambroxol with ammonium acetate and sodium hypochlorite significantly attenuated this enhanced gastric carcinogenesis. Results also revealed that ambroxol scavenged monochloramine. Because monochloramine is closely related to Helicobacter pylori associated gastric carcinogenesis, these findings suggest that ambroxol may prevent H. pylori-associated gastric carcinogenesis. PMID- 11403916 TI - Inhibition by dietary dibenzoylmethane of mammary gland proliferation, formation of DMBA-DNA adducts in mammary glands, and mammary tumorigenesis in Sencar mice. AB - Dibenzoylmethane (DBM) is a minor constituent of licorice and a beta-diketone analogue of curcumin. Feeding 1% DBM in the diet to Sencar mice during both the initiation and the post-initiation periods strongly inhibited 7,12 dimethylbenz[a]anthracene (DMBA)-induced mammary tumor multiplicity and mammary tumor incidence by 97%. In further in vivo studies to elucidate the possible mechanisms of the inhibitory action of DBM, feeding the 1% DBM in the AIN-76A diet to immature Sencar mice for 4-5 weeks decreased the uterine wet weight by 43%, inhibited the proliferation rate of mammary gland epithelial cells by 53%, uterine epithelium by 23%, and uterine stroma by 77%, when mice were killed during the first estrus phase of estrous cycle. In addition, feeding 1% DBM in the diet to Sencar mice at 2 weeks before, during and 1 week after DMBA treatment (intubation of 1 mg DMBA per mouse once a week for 5 weeks) inhibited formation of total DMBA-DNA adducts in mammary glands by 72% using a post-32P-labeling assay. Thus, feeding 1% DBM diet to Sencar mice inhibited formation of DMBA-DNA adducts in mammary glands and lowered the proliferation rate of the mammary gland in vivo. These results may explain the strong inhibitory actions of dietary DBM on mammary carcinogenesis in mice. PMID- 11403917 TI - Inverse association between cyclooxygenase-2 overexpression and microsatellite instability in gastric cancer. AB - This study examined the association between cyclooxygenase-2 (COX-2) overexpression and microsatellite instability (MSI) in gastric cancer. COX-2 expression was assessed by immunohistochemistry and scored in a semi-quantitative manner whereas MSI status was characterized by nine microsatellite markers. The clinicopathological features of cancers including survival data were analyzed. Of the 109 gastric cancers studied, COX-2 overexpression and high level of MSI (MSI H) was detected in 64.2 and 22.0% cases respectively. Gastric tumors with MSI-H phenotypes had significantly lower level of COX-2 expression levels when compared to MSI-L and MSS tumors (P=0.002). Moreover, COX-2 overexpression was associated with tumor invasion beyond submucosa (P=0.045) and there was a trend favoring better survival in gastric cancers without COX-2 overexpression (P=0.07). The results from this study suggest that gastric cancer with microsatellite instability or COX-2 overexpression present with diverse clinicopathological features. PMID- 11403918 TI - Pro-apoptotic interactions between XK469 and the peripheral benzodiazepine receptor. AB - XK469 (2-[4-(7-chloro-2-quinoxalinyloxy) phenoxy]propionic acid) is a new anti tumor agent with substantial activity against several drug-resistant cell lines. Using murine leukemia L1210 cells in culture, we found the chiral R(+) form of XK469 to be substantially more cytotoxic than the S(-) form, while the herbicide analog 'Assure' was essentially inactive. The cytotoxic response to these agents was accompanied by apoptosis, and was found to be correlated with drug binding to the peripheral benzodiazepine receptor in cell culture, suggesting that receptor binding may be a factor in drug-induced cytotoxicity. PMID- 11403919 TI - Functional maturation of dendritic cells by exposure to CD40L transgenic tumor cells, fibroblasts or keratinocytes. AB - Tumor antigen pulsed dendritic cells (DCs) can induce anti-tumor immunity. We studied strategies for the reliable generation of such a tumor vaccine by functional maturation of DCs via interaction of CD40 with its ligand (CD40L, CD154). Exposure of immature DCs to CD40L transgenic cells, soluble recombinant human CD40L molecules or lipopolysaccharide induced expression of the co stimulatory molecules, CD80 and CD86, and supported an allogeneic mixed leukocyte reaction. In contrast, the release of IL-12, an important mediator of anti-tumor immunity, and antigen-specific expansion and IFNgamma secretion of lymphocytes, was strongly triggered only by DCs exposed to CD40L transgenic cells. PMID- 11403920 TI - Chemoprevention of NMU-induced rat mammary carcinoma with the combination of melatonin and 9-cis-retinoic acid. AB - In experimental trials using the N-nitroso-N-methylurea (NMU)-induced rat mammary tumor model, a significant decrease in tumor incidence (to 5%) was observed in rats treated with melatonin and 9-cis-retinoic acid (9 cRA) compared to controls (55%). Although 9cRA alone decreased tumor incidence to 26%, this response did not reach statistical significance. Tumor incidence was significantly inhibited to 20% in the animals that received melatonin and 9cRA on alternating days. Latency to tumor onset was prolonged in animals receiving either of the combination treatments compared with controls, and tumor multiplicity was also significantly decreased. PMID- 11403921 TI - Cyclin D1 as a proliferative marker regulating retinoblastoma phosphorylation in mouse lung epithelial cells. AB - Elevations in cyclin D1 content increase the phosphorylation status of retinoblastoma (Rb) protein to encourage cell cycle transit. We sought to determine if cyclin D1 content could be used as an index of cell proliferation in mouse lung epithelia following growth manipulations in vitro and in vivo. Rb protein concentration was high in 82-132 and LM2, two fast-growing neoplastic mouse lung epithelial cell lines. The hyperphosphorylated form of Rb predominated in these two cell lines, while Rb in slower-growing cell lines was predominantly hypophosphorylated. Consistent with this, more cyclin D1 protein was expressed in the fast-growing cell lines than in slower-growing cells. We therefore tested whether cyclin D1 content varied with growth status. The amount of cyclin D1 decreased upon serum removal coincident with growth inhibition and then increased upon serum re-addition which stimulated resumption of proliferation. This correlation between cyclin D1 content and growth status also occurred in vivo. Cyclin D1 content increased when lungs underwent compensatory hyperplasia following damage caused by butylated hydroxytoluene administration to mice and in lung tumor extracts as compared with extracts prepared from uninvolved tissue or control lungs. We conclude that elevated cyclin D1 levels account, at least in part, for the hyperphosphorylation of Rb in neoplastic lung cells, and are associated with enhanced lung growth in vitro and in vivo. PMID- 11403922 TI - Heat shock protein 27 was up-regulated in cisplatin resistant human ovarian tumor cell line and associated with the cisplatin resistance. AB - To understand the molecular basis for failure of cisplatin (CDDP) based chemotherapy, we compared gene expressions between CDDP sensitive and resistant ovarian tumor cell line, 2008 and 2008/C13*5.25, by mRNA differential display. We detected both up-regulated and down-regulated bands in the resistant cell and found some of them to be positive on Northern blotting. DNA sequencing revealed one to be mitochondrial heat shock protein 75. We found that HSP27 and HSP70 were also up-regulated in the resistant cell by Western blotting. Further, transient transfection with the HSP27 sense gene made the sensitive cell more resistant, while transient transfection with the antisense gene made it more sensitive. PMID- 11403923 TI - 2,5-Di-tert-butyl-1,4-hydroquinone enhances cell transformation accompanied by an increase in intracellular free calcium ion concentration. AB - Two hydroquinones with similar structures, 2,5-di-tert-butyl-1,4-hydroquinone (DTBHQ) and 2-tert-butyl-1,4-hydroquinone (MTBHQ), are used as antioxidants in the environment. DTBHQ and MTBHQ were examined for their ability to induce cell transformation using BALB/3T3 cells. DTBHQ at concentrations of 2.5-15 microM enhanced cell transformation initiated by a subthreshold dose (0.75 microM) of 3 methylcholanthrene (MCA) in a two-stage cell transformation assay, which mimics a two-stage carcinogenicity test in experimental animals. Because DTBHQ is known to act as a calcium ion mobilizing agent in other cells, we examined the effects of DTBHQ on intracellular free calcium ion concentration ([Ca(2+)]i) in BALB/3T3 cells. DTBHQ elevated [Ca(2+)]i with a dose dependency similar to that of its enhancing effect on the MCA-initiated cell transformation. MTBHQ neither enhanced cell transformation nor induced increase of [Ca(2+)]i. Aberrant calcium signaling produced by DTBHQ might contribute to the enhancement of MCA-initiated transformation in BALB/3T3 cells. PMID- 11403924 TI - Expression of the prothymosin alpha mRNA correlated with that of N-myc in neuroblastoma. AB - Neuroblastoma is the most common malignant solid cancers in early childhood. Overexpression of the proto-oncogene, N-myc, has been reported to be correlated with more malignant course of the disease. Prothymosin alpha, a cellular proliferation-associated gene, is reported to be a target of myc and elevated in several malignant cells and tissues. Expression of prothymosin alpha and N-myc messenger RNAs were evaluated by real-time reverse transcription polymerase chain reaction (RT-PCR) assay in 18 tumor samples from neuroblastoma using LightCycler. The data was analyzed in reference to clinicopathological factors. There was a tendency that higher prothymosin alpha transcripts levels in the tumor samples from younger patients (<1year.) when compared to the older group (>1 year.) (P=0.0845). There was no relationship between prothymosin alpha gene expression and gender (P=0.3029), mass screening case or not (P=0.3007), or stage. The prothymosin alpha mRNA expression levels were correlated with N-myc mRNA levels (P=0.006). Thus we suggest that prothymosin alpha plays an active role as a target of N-myc in neuroblastoma. PMID- 11403925 TI - Expression of a tumor-associated antigen RCAS1 in hepatocellular carcinoma. AB - RCAS1 has been reported as a tumor-associated antigen in uterine and ovarian carcinomas. In vitro studies on RCAS1 indicated that it might function as an apoptosis-inducing factor since binding between RCAS1 and its receptor induced apoptosis in receptor-expressing cells. In this study, 68 surgically resected samples of hepatocellular carcinoma (HCC) were prepared and RCAS1 expression was examined immunohistochemically, because RCAS1 was also positive in all HCC cell lines tested. Clinical and pathological parameters were then compared between RCAS1-positive and -negative HCC cases. As a result, RCAS1 is expressed in 26.5% of HCC cases and vascular invasion is observed at a much higher rate in the RCAS1 positive cases (72.2%) than in RCAS1-negative cases (24.0%). RCAS1 is not an antigen specific for gynecological cancers. In HCC cases, the RCAS1-positive percentage is not high, however, RCAS1-positive HCCs exhibited a trend towards invasive character. PMID- 11403926 TI - Long-term impairment of social memory in the rat after social defeat is not restored by desglycinamide-vasopressin. AB - Repeated social defeat followed by individual housing caused a long-term impairment of social memory in male rats. Social memory, as assessed in the social discrimination test using an intertrial interval of 3 min, was impaired for at least 8 weeks after the social defeat experience. Since social memory of male rodents depends on proper functioning of the sexually dimorphic vasopressin system, it was investigated whether a centrally active vasopressin fragment could restore the impaired social memory. Subcutaneous administration of 6 microg/kg of the vasopressin fragment desglycinamide-vasopressin (VP1-8) 40 days after social defeat slightly improved social memory in both control and socially defeated rats. It is concluded that social defeat followed by individual housing caused a long-term impairment of social memory, which was not restored by treatment with VP1-8. PMID- 11403927 TI - Effects of transient, mild mood states on semantic memory organization and use: an event-related potential investigation in humans. AB - The effects of transient mood states on semantic memory organization and use were investigated using event-related potentials. Participants read sentence pairs ending with (1) the most expected word, (2) an unexpected word from the expected semantic category, or (3) an unexpected word from a different (related) category; half the pairs were read under neutral mood and half under positive mood. Under neutral mood, N400 amplitudes were smallest for expected items and smaller for unexpected items when these came from the expected category. In contrast, under positive mood, N400 amplitudes to the two types of unexpected items did not differ. Positive mood seemed to specifically facilitate the processing of distantly-related, unexpected items. The results suggest that transient mood states are associated with dynamic changes in how semantic memory is used on line. PMID- 11403928 TI - Increased Bcl-w expression following focally evoked limbic seizures in the rat. AB - Control of seizure-induced neuronal death may involve members of the Bcl-2 family of cell death regulating proteins. Bcl-w is a newly described anti-apoptotic member of this family that may confer neuroprotective effects. We therefore investigated Bcl-w expression in rat brain following focally evoked limbic seizures. Seizures were induced by unilateral microinjection of kainic acid into the amygdala of the rat and terminated after 40 min by diazepam. Constitutive Bcl w expression was detected by Western blotting and immunohistochemistry. Bcl-w expression was increased 4-72 h following seizures within the injured hippocampus. Immunohistochemistry determined Bcl-w was predominantly expressed in neurons and seizures increased Bcl-w immunoreactivity within piriform cortex and surviving regions of the injured hippocampus. These data suggest Bcl-w may be involved in the modulation of seizure-induced brain injury. PMID- 11403929 TI - Nerve growth factor and neurotrophin-3 promote chemotaxis of mouse macrophages in vitro. AB - Microchemotaxis chambers were used to explore macrophage chemotaxis in response to the neurotrophins, nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT-3). Both NGF and NT-3, but not BDNF, promoted macrophage chemotaxis that was receptor mediated and dependent on protein phosphorylation. These in vitro observations point to novel roles for neurotrophins that are present in nerve prior to and immediately after injury. PMID- 11403930 TI - Interaction of local anaesthetic agents with the endogenous norepinephrine transporter in SH-SY5Y human neuroblastoma cells. AB - Use of intravenous guanethidine for the treatment of complex regional pain syndrome type I is of variable efficacy. Guanethidine injection is painful, so local anaesthetic is co-administered. We hypothesize that local anaesthetic inhibits uptake of guanethidine and hence reduces its efficacy. In this study we have examined the effects of a range of local anaesthetic agents on the uptake of [3H]norepinephrine ([3H]NE) (as a surrogate for guanethidine) and the binding of [3H]nisoxetine to the NE transporter in cultured SH-SY5Y human neuroblastoma cells. All local anaesthetic agents inhibited NE uptake with a rank order cocaine>tetracaine>procaine(esters), dibucaine > bupivacaine > prilocaine > lidocaine (amides). In addition all anaesthetic agents displaced [3H]nisoxetine with a rank order cocaine > tetracaine > dibucaine > procaine > prilocaine > bupivacaine > lidocaine. There was a positive correlation between [3H]NE uptake and [3H]nisoxetine binding. Our data suggest that when local anaesthetic and guanethidine are co-administered the former may reduce uptake of the latter and hence reduce the clinical efficacy of guanethidine. PMID- 11403931 TI - Complement activation by neurofibrillary tangles in Alzheimer's disease. AB - Brain inflammation is widely documented to occur in Alzheimer's disease (AD), but its sources are still incompletely understood. Here, we present in vitro and in situ evidence that, like amyloid beta peptide (Abeta), tau, the major protein constituent of the neurofibrillary tangle, is a potent, antibody-independent activator of the classical complement pathway. Complement activation, in turn, is known to drive numerous inflammatory responses, including scavenger cell activation and cytokine production. Because Abeta deposits and extracellular tangles are present from early preclinical to terminal stages of AD, their ability to activate complement provides a ready mechanism for initiating and sustaining chronic, low-level inflammatory responses that may cumulate over the disease course. PMID- 11403932 TI - Enriched environment influences brain-derived neurotrophic factor levels in rat forebrain after focal stroke. AB - Tissue levels of brain-derived neurotrophic factor (BDNF) protein were studied using enzyme immunoassay in different forebrain regions in the ipsi- and contralateral hemispheres of rats housed under enriched or standard conditions after the middle cerebral artery ligation. BDNF levels in the ipsilateral to ligation side was significantly higher only in the frontal cortex of standard as compared to enriched rats. However, BDNF overall was more abundant in standard than in enriched group. In addition, BDNF levels detected in the hippocampus and frontal cortex on the ischemic side of standard rats was higher as compared to contralateral side. The present study shows that housing conditions after permanent middle cerebral artery ligation leads to differential regulation of BDNF protein levels in forebrain regions which might have important implication for post-ischemic recovery. PMID- 11403933 TI - Altered laminar distribution of hippocampal zinc in mutant mice lacking neuronal nitric oxide synthase. A histochemical study. AB - The recently discovered role of nitric oxide (NO) in the accumulation of zinc in the central nervous system encouraged us to compare the distribution of zinc in the hippocampal formation and other brain regions of nitric oxide synthase knockout with wild-type mice. The histochemical Timm's staining technique was used. While no changes in the patterns of zinc staining were observed in extrahippocampal brain areas, lamina-specific differences appeared in the distribution of the metal in hippocampi of normal and mutant mice. Less Timm's stainable zinc was detected in the neuropil of strata oriens and radiatum and a loss of Timm's-stained infrapyramidal fibers in knockout mice. The hilar region as well as the suprapyramidal and intrapyramidal blades of the mossy fiber system did not differ between mutant and wild-type mice. Our data show that life-long withdrawal of neuronal-derived NO has effects on the accumulation of zinc in the hippocampal formation of mice. PMID- 11403934 TI - High specificity of human orexin receptors for orexins over neuropeptide Y and other neuropeptides. AB - Orexin- and neuropeptide Y-ergic systems show physiological interaction in the regulation of appetite. In this study we investigate the postulated effect of neuropeptide Y (NPY) and other peptides directly on orexin OX1 and OX2 receptors. None of the tested peptides (NPY-variants, secretin, alpha-melanocortin, pancreatic polypeptide or pituitary adenylyl cyclase-activating peptide-38) induced any Ca2+ elevation at the concentrations up to 300 nM (human NPY, secretin and alpha-melanocortin up to 1 microM). Orexin-A- and -B-mediated Ca2+ elevations were completely unaffected by the peptides. In binding assays, human NPY, secretin and alpha-melanocortin at 1 microM did not induce any displacement of 0.1 nM [125I]orexin-A. Thus, in contrast to the previously reported result on orexin-A binding, our results demonstrate that NPY does not directly interact with orexin receptor in intact cellular settings. PMID- 11403935 TI - Histamine production by cultured microglial cells of the mouse. AB - We previously reported that cells other than mast cells or neurons could synthesize histamine in response to lipopolysaccharide (LPS) or interleukin 1beta in the rat brain. To identify the responsible cells, we examined histidine decarboxylase (HDC) activity and the expression of HDC mRNA in GMI 6-3 mouse microglial cells. Both the activity and mRNA for HDC in GMI 6-3 cells were induced by LPS treatment, and the induction was sensitive to calmodulin-dependent kinase II inhibitor, KN62. These findings indicate that microglia is a third cell type producing histamine in the brain. PMID- 11403936 TI - A quantitative study on the expression of synapsin II and N-ethylmaleimide sensitive fusion protein in schizophrenic patients. AB - The application of DNA array technology to schizophrenic studies enabled us to assess molecular features of this disease. The expression of synapsin II and N ethylmaleimide-sensitive fusion protein (NSF) mRNAs is reported to decrease in the prefrontal cortex of these patients. We attempted to reproduce this result with two distinct approaches. With high quality samples, mRNA and protein levels for synapsin II and NSF were measured by real-time polymerase chain reaction and by immunoblotting. Both experiments led to the same conclusion: The expression of these presynaptic markers is not altered significantly in the prefrontal cortex of our schizophrenic samples, compared to that in control subjects. These observations suggest that the neurochemical impairments of synapses reported in schizophrenia are not evident for all presynaptic markers and needs to be re evaluated at molecular levels. PMID- 11403937 TI - Fighting among C57BL/6J mice and its implications for [3H]8-hydroxy-N, N-dipropyl 2-aminotetralin binding in various brain regions. AB - The binding of [3H]8-hydroxy-N, N-dipropyl-2-aminotetralin ([3H] 8-OH-DPAT) has been studied in various brain regions of male mice with the experience of repeated victory (winners) or defeat (losers) gained over 10 (T10) and 20 (T20) days of daily agonistic confrontations. The Bmax of [3H]8-OH-DPAT binding sites was found to be decreased in the hippocampus of the T20 winners when compared to the T10 winners or the control mice, and increased in the amygdala of the T20 losers. The Kd was increased in the amygdala of the T20 losers when compared to the animals of any other experimental group. No change in [3H]8-OH-DPAT binding was observed in the midbrain or the hypothalamus of the winners or losers. It was concluded that the 20-day agonistic confrontations, arranged in order to categorize the mice as winners or losers, had opposing effects on [3H]8-OH-DPAT binding. PMID- 11403938 TI - Reversal by NS-7, a neuroprotective compound, of the decrease in transcription factor CREB mRNA expression in rat brain after permanent middle cerebral artery occlusion. AB - The effect of a neuroprotective agent NS-7 on changes in mRNA expressions for cyclic AMP responsive element binding protein (CREB) and several neurotrophins was examined in the rat cerebral cortex after permanent middle cerebral artery occlusion (MCAO). Significant reduction in mRNA expressions for CREB was observed at 24h after MCAO. NS-7 (0.5mg/kg), when injected at 6h after MCAO, significantly reversed the decreased expression for CREB mRNA. In addition, the mRNA expression for basic fibroblast growth factor (bFGF) was also significantly enhanced by NS-7 in MCA-occluded but not in sham-operated rats. On the other hand, the mRNAs for interluekin-6 and inducible-type nitric oxide synthase were markedly induced in the cerebral cortex of MCA-occluded rats, which was not significantly reversed by NS-7. Therefore, it is suggested that the reversal of decrease in CREB mRNA and concomitant increase in mRNA expression for bFGF may contribute to the neuroprotective action of NS-7. PMID- 11403939 TI - Developmental differences in cortical and hippocampal vulnerability to intermittent hypoxia in the rat. AB - Obstructive sleep apnea is characterized by intermittent hypoxic events during sleep, and is associated with substantial neurocognitive morbidity, particularly in children. Intermittent hypoxia (IH) leads to increases in apoptosis in the cortex and hippocampus of the adult rat, peaking at 48 h of exposure. To examine whether the susceptibility to IH exhibits developmental differences, rats were exposed to 48 h of IH at ages 2, 5, 10, 15, 20, 25, 30, 60, and 120-day postnatally, and apoptosis was determined by terminal deoxy-nucleotidyl transferase-mediated in situ end labeling and immunohistochemical staining for single-stranded DNA. Although IH induced apoptosis at all postnatal ages, smaller increases were apparent in 2 and 5-day old (P < 0.01 vs. any other age) while peak apoptosis occurred at 10-25 days (P < 0.001 vs. 30, 60, and 120 days). We conclude that a unique window of vulnerability to IH is present in the cortex and hippocampus during post-natal maturation, and may underlie the high frequency of neurobehavioral deficits associated with obstructive sleep apnea in children. PMID- 11403940 TI - Transient massive DNA fragmentation in nervous system during the early course of a murine neurodegenerative disease. AB - In neurodegenerative diseases, such as Alzheimer's disease or HIV encephalitis, neuronal DNA fragmentation has been observed at unexpected high frequencies, without definitive evidence for activation of an irreversible apoptotic pathway. The wobbler mouse is a suggested genetic model of neurodegenerative disease. The mutant mouse develops normally until the fourth week of age when atrophy and weakness of forelimb muscles become apparent. There is a slow progression of the disease and wobbler mice may survive for several months. Spinal cord examination reveals the presence of several motoneurons with perikaryal vacuolar degeneration. In this study, we observed, using terminal dUTP nick-end-labelling staining in mutant spinal cord sections, a massive although very transient DNA fragmentation in different cell types, including glial cells and motoneurons, before the apparition of any clinical symptoms. In older wobbler mice, this DNA fragmentation had completely disappeared and the majority of motoneurons survived. To our knowledge, this is the first example of a massive and transient DNA fragmentation in the central nervous system during the early course of a neurodegenerative disease. PMID- 11403943 TI - Predominant functional activity of the large, neutral amino acid transporter (LAT1) isoform at the cerebrovasculature. AB - In this study, we identify the predominant functional expression of the large, neutral amino acid transporter (LAT1) isoform at the blood-brain barrier (BBB). An in situ rat brain perfusion technique allowed perfusion of the radiotracer [(14)C]-L-Leu (a ligand for both LAT1 and LAT2) alone or competed with excess concentration of either LAT1 or LAT2 specific amino acids. The LAT2 specific amino acid, [(14)C]-L-Asn, was perfused alone or with excess concentration of various amino acids. The brain uptake of [(14)C]-L-Leu was not significantly inhibited by LAT2 specific amino acids, but was inhibited significantly (up to 90%) by the LAT1 specific amino acid, D-Met. L-Asn did not demonstrate saturable brain uptake. These data clearly demonstrate that LAT1 is the functionally predominant isoform expressed at the BBB which is responsible for brain uptake of large, neutral amino acids. In addition, the functional activity of cerebrovascular LAT2 is insignificant, or absent. PMID- 11403944 TI - Levetiracetam inhibits the high-voltage-activated Ca(2+) current in pyramidal neurones of rat hippocampal slices. AB - The effect of the new antiepileptic drug levetiracetam (LEV; KEPPRA) on the neuronal high-voltage-activated (HVA) Ca(2+) current was investigated on pyramidal neurones, visually identified in the CA1 area of rat hippocampal slices. Nystatin-perforated patch clamp recordings were made under experimental conditions designed to study HVA Ca(2+) currents. The HVA current, activated by steadily increasing voltage-ramps, was reversibly eliminated by Cd(2+) and depressed by either nimodipine, or omega-Conotoxin GVIA. After 30 min perfusion of the slices with LEV 32 microM, the current decayed to 55+/-9% (mean+/-SEM; n=9) of the initial value, which is significantly (P<0.05, two-tailed t-test) lower than the rundown to 84+/-10% in a control group (n=10) of neurones. The limited, but significant depression of the neuronal HVA Ca(2+) current, produced by LEV at a clinically relevant concentration, might contribute to the antiepileptic action of the drug. PMID- 11403945 TI - Allopurinol suppresses para-nonylphenol and 1-methyl-4-phenylpyridinium ion (MPP(+))-induced hydroxyl radical generation in rat striatum. AB - We recently demonstrated that para-nonylphenol, an environmental estrogen-like chemical, enhances hydroxyl radical (*OH) generation in the rat striatum. In the present study we have examined whether para-nonylphenol enhanced 1-methyl-4 phenylpyridinium ion (MPP(+))-induced *OH generation in the rat striatum using a microdialysis technique. Para-nonylphenol significantly enhanced MPP(+)-induced *OH generation. Further, we studied the effect of allopurinol, a xanthine oxidase inhibitor, on para-nonylphenol and MPP(+)-induced *OH generation. Allopurinol significantly suppressed para-nonylphenol and MPP(+)-induced *OH generation. The results indicate that para-nonylphenol enhanced *OH generation based on superoxide anion production, and allopurinol may have preventive effect on para nonylphenol and MPP(+)-induced *OH generation. PMID- 11403946 TI - Effect of finasteride on behavioural arousal and somatosensory evoked potentials in fetal sheep. AB - This study examines the effect of inhibiting the synthesis of gamma-aminobutyric acid(A) (GABA(A)) agonist steroids on behavioural activity and somatosensory evoked potentials (SEP) in late gestation fetuses. Pregnane steroid production was suppressed by infusion of the 5alpha-reductase inhibitor, finasteride in chronically catheterised fetal sheep, 130-135 days gestation. Finasteride treatment (160 mg in 10 ml of vehicle over 2 h) significantly increased the incidence of fetal arousal during the period 4-10 h after commencing the infusion (P<0.05, n=6), whereas other behavioural parameters were not effected. In three of four animals, finasteride produced an increase in the amplitude of the N22 peak of the SEP during high voltage electrocortical activity. We conclude that suppression of pregnane steroid synthesis, by inhibition of the 5alpha-reductase enzyme, increases arousal activity in the fetus which is consistent with a reduction in GABA(A) receptor mediated inhibition. PMID- 11403947 TI - Tiagabine treatment and DNA damage in rat astrocytes: an in vitro study by comet assay. AB - We studied in vitro the effects of Tiagabine on genomic DNA of cortical rat astrocytes. To evaluate DNA damage, we used a relatively simple technique called Single Cell Gel Electrophoresis or Comet assay. Tiagabine was dissolved in culture medium and added at concentration of 1, 10, 20 and 50 microg/ml on 12-day old cultured astrocytes. In presence of 1 and 10 microg/ml of Tiagabine, no DNA damage was observed after 48 h of treatment. A moderate DNA damage was instead observed for cells exposed to 20 microg/ml of antiepileptic drug. Finally, DNA fragmentation was more evident after treatment with 50 microg/ml of Tiagabine. We conclude that Tiagabine, at the usual recommended doses, does not appear to influence negatively the cortical rat astrocytes, inducing DNA fragmentation only at very high concentrations. PMID- 11403948 TI - Differential regulation of striatal G protein levels following 1-methyl-4-phenyl 1,2,3,6-tetrahydropyridine administration in C57 BL/6 mice. AB - The dopaminergic neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is known to produce a severe Parkinsonian state in both humans and animals. Unlike idiopathic Parkinson's disease, however, most MPTP models show some degree of behavioral recovery with time. Here we report that stimulatory G proteins are differentially regulated in the striatum of C57 BL/6 mice following systemic MPTP administration. As measured by Western blotting, the striatal stimulatory G proteins Gs and Golf were reduced by 20% and 25% at 10 days following cessation of MPTP treatment, despite a significant impairment in striatal dopamine levels (<90% reduction). Conversely, Gs and Golf levels were upregulated by 15% and 30% at 10 months following MPTP withdrawal. No change was observed in striatal inhibitory G proteins or any cortical G protein at any time post-treatment. These results suggest that G protein upregulation may play a role in mediating behavioral recovery following MPTP administration. PMID- 11403949 TI - Characterization of novel benzodiazepine ligands in Spodotera frugiperda (Sf-9) insect cells. AB - The goals of the present work were to characterize the binding profile of nine benzodiazepine ligands in Spodotera frugiperda (Sf-9) insect cells expressing specific gamma aminobutyric acid (A) (GABA(A)) receptor subunit combinations and compare the affinities to those for the receptors in the rat cerebellum. Three recombinant baculovirus constructs, each harboring a different GABA(A) receptor subunit, were introduced into insect cells by simultaneous infection. Saturation and competition binding assays were carried out in membranes from Sf-9 cells infected with either alpha1beta2gamma2 or alpha6beta2gamma2 subunit combinations. The affinities of the ligands to the alpha1beta2gamma2 or alpha6beta2gamma2 receptors expressed in Sf-9 cells were similar to the affinities previously determined for the alpha1 or alpha6 subunit-containing GABA(A) receptors in the rat cerebellum, respectively, thus confirming the previously assigned receptor types in the cerebellum. PMID- 11403950 TI - Increased variability of paced finger tapping accuracy following repetitive magnetic stimulation of the cerebellum in humans. AB - Imaging and lesion studies suggest that the cerebellum is involved in the self generation of timed motor responses. Using repetitive transcranial magnetic stimulation (rTMS), we studied the effects of transient disruption of the lateral or medial cerebellum on a paced-finger-tapping task (PFT). Results show greater variability on the PFT task following a 5 min train of 1 Hz rTMS to the medial cerebellum. Magnetic stimulation of the lateral cerebellum or motor cortex, and sham stimulation, had no effect on performance. Expanding the results of neuroimaging studies, these data show the causal link between activity in the medial cerebellum and the production of timed movements. This is the first demonstration of the feasibility of transiently disrupting the cerebellum by rTMS and inducing behavioral effects. This method of 'virtual lesions' can expand the study of the role of the cerebellum in motor control and cognition. PMID- 11403951 TI - Elevation of prefrontal acetylcholine is related to the extinction of learned behavior in rats. AB - To investigate whether changes occur in acetylcholine (ACh) levels in the rat medial prefrontal cortex (mPFC) during initial lever-press extinction training, in vivo microdialysis was used to measure mPFC ACh. Elevated ACh was found during this training period. Furthermore, this elevation significantly correlated with the number of responses found in the re-training session the next day, but not with that in the initial training. These results suggest that the mPFC ACh elevation during the initial training period enhances the progress of lever-press extinction across sessions. PMID- 11403952 TI - Involvement of cyclin dependent kinase5 activator p25 on tau phosphorylation in mouse brain. AB - P35 or its truncated fragment p25 is required for cyclin dependent kinase (Cdk)5 activation. It has been reported that p25 is accumulated in the brain of Alzheimer's disease (AD) patients and that p25/Cdk5 induces high phosphorylation of tau and apoptosis in cultured neurons (Nature 402 (1999) 615). Our investigation of AD brain did not show specific accumulation of p25. Exposure to Ca ionophore (A23187) at 10(-6) M induced p25 accumulation in rat primary hippocampal neurons, causing neuronal death without showing hyperphosphorylation of tau. Transgenic mice expressing p25 showed the accumulation of p25 but neither hyperphosphorylation of tau nor neuronal death was shown in these mice. The feature of these mice was the progression of cell growth in pituitary gland. These results suggest that overexpression of p25 lead to the activation of cell cycle but not to the direct phosphorylation of tau. PMID- 11403953 TI - Mutant mice lacking the cholecystokinin2 receptor show a dopamine-dependent hyperactivity and a behavioral sensitization to morphine. AB - Cholecystokinin2 (CCK2) receptor-deficient mice were used to analyze the in vivo function of CCK2 receptor and especially the incidence of this gene invalidation on enkephalinergic and dopaminergic systems. Hyperlocomotor activity of CCK2 receptor-deficient mice was suppressed by a selective D2 antagonist but not by a D1 antagonist. Injection of amphetamine induced a hyperlocomotor activity in both groups of mice while mutant mice were less sensitive to cocaine. Administration of 6 mg/kg of morphine once every 2 days for 5 days significantly (P<0.05) enhanced motor activity the last day compared to the first day, only in CCK2 receptor-deficient mice. These results emphasize the role of CCK2 receptors in counteracting the effects of dopaminergic systems and suggest that CCK2 receptor invalidation could lead to a slight behavioral sensitization. PMID- 11403954 TI - Spontaneously active GABAergic interneurons in the subfornical organ of rat slice preparations. AB - Inhibitory postsynaptic currents (IPSCs) were recorded from subfornical organ (SFO) neurons in slice preparations of rats, using whole-cell voltage clamp techniques. Some SFO neurons showed bimodal distributions in amplitude with the large and small IPSCs. The large IPSCs vanished in the tetrodotoxin perfusion medium, but the small did not. Both sizes of the IPSCs were completely abolished by application of bicuculline and picrotoxin. Further subpopulation of SFO neurons with the bimodal distributions showed intermittent bursts of the large IPSCs. Immunohistochemical approach revealed existence of gamma-aminobutyric acid (GABA)-immunoreactive neurons and axons in the SFO. These suggest that spontaneously-active and intermittently-burst-firing GABA interneurons affect other SFO neurons in slice preparations of rats. PMID- 11403955 TI - Sequence, protein expression and extracellular-regulated kinase association of the hyaladherin RHAMM (receptor for hyaluronan mediated motility) in PC12 cells. AB - Differentiation of PC12 cells by nerve growth factor (NGF) or fibroblast growth factor-2 (FGF2) is dependent on signaling mediated by extracellular regulated kinase (ERK). We investigated the involvement of receptor for hyaluronan mediated motility (RHAMM) in this signaling pathway. A single RHAMM 3.2 kb transcript was detected in PC12 RNA. Reverse transcriptase-polymerase chain reaction generated a 2141 bp cDNA that had identical sequence to rat brain RHAMM and showed no evidence of alternate splicing. Several RHAMM species were identified by Western blotting. Immunohistochemistry showed RHAMM localization to the cytoskeleton, neurites and growth cones. Following stimulation of PC12 cells with NGF or FGF2 RHAMM was co-immunoprecipitated by phosphorylation-specific anti-ERK antibodies, indicating a role for RHAMM in ERK signaling in PC12 cells. PMID- 11403956 TI - Huperzine A and donepezil protect rat pheochromocytoma cells against oxygen glucose deprivation. AB - Huperzine A (HupA) and donepezil, two novel selective acetylcholinesterase inhibitors available for Alzheimer's disease, were tested for their ability to alleviate injury from oxygen-glucose deprivation (OGD) in the rat pheochromocytoma line PC12 cells. OGD for 30 min triggered death in more than 50% of cells, along with major changes in morphology and biochemistry including elevated levels of lipid peroxide, superoxide disamutase activity and lactate. Cells pretreated for 2 h with HupA or donepezil showed improved survival and reduced biochemical and morphologic signs of toxicity (statistically significant over the range from 10 microM down to 1.0 and 0.1 microM, respectively). Our results indicated that HupA and donepezil protected PC12 cells against OGD induced toxicity, most likely by alleviating disturbances of oxidative and energy metabolism. PMID- 11403957 TI - Transplantation of adult rat hippocampus-derived neural stem cells into retina injured by transient ischemia. AB - Neural stem cells are capable of differentiating along multiple central nervous system cell-type lineages, and their use as graft material has provided new strategies for the treatment of neuronal damage. We transplanted adult rat hippocampus-derived neural stem cells into eyes of adult rats that underwent ischemia-reperfusion injury. As control, the cells were also injected into normal rats eyes without ischemic insult. The rats were sacrificed at 1, 2, 4, and 8 weeks, and the eyes were examined histochemically. In eyes with the insult, the transplanted cells were well integrated into the host retinas and expressed Map2ab. In the control, none of the cells migrated into the retina. These results suggest that neural stem cells may be used as donor cells for transplantation to repair ischemic-injured retina. PMID- 11403958 TI - The cerebrovascular response to elevated potassium--role of nitric oxide in the in vitro model of isolated rat middle cerebral arteries. AB - We investigated the role of nitric oxide (NO) in the vascular response to high extraluminal K(+)-concentrations in the in vitro model of isolated rat middle cerebral arteries (MCA). Under control conditions, rat MCA dilated at 20, 30, 40 and 60 mM K(+). At 80 mM K(+), a slight vasoconstriction occurred. The unspecific NO synthase (NOS)-inhibitor L(omega)-nitro-L-arginine (L-NNA) increased the resting tone at 3 mM K(+) by 31+/-5% (P<0.01). While the vasodilatative effect of 20 mM K(+) was unaffected by L-NNA, NOS-inhibition resulted in vasoconstriction at > or = 40 mM K(+) (P<0.01). In presence of L-NNA, the basal vessel diameter was restored by either the NO-donor S-nitroso-N-acetylpenicillamine (SNAP) or the cell-permeable guanosine-3',5'-cyclic monophosphate (cGMP) analogue 8-Br-cGMP. Co application of L-NNA with either SNAP or 8-Br-cGMP resulted in partial restitution of the vasodilatative effect of 40 mM K(+), respectively. In presence of the soluble guanylyl cyclase inhibitor 1 H-[l,2,4]oxadiazolo[4,3-a]quinoxalin 1-one (ODQ), the vascular response to 40 mM K(+) was abolished. Our findings together with findings from the literature indicate a modulator role of NO at K(+) > or = 40 mM K(+), involving a cGMP-dependent mechanism. PMID- 11403959 TI - Sleep and memory deficits in the rat produced by experimental infection with Trypanosoma cruzi. AB - We investigated whether the infection with Trypanosoma cruzi in rats could produce functional alterations of the central nervous system. The experimental group received an injection of 150,000 trypomastigotes / rat, whereas the control group received a saline injection. Spontaneous alternation behavior (SAB) tests and sleep-wake cycle recordings were obtained at the end of the parasitaemia. Results showed that the infected animals had significant sleep impairments, as denoted by an increase in the number of wake periods and a reduction of rapid eye movement sleep amount. SAB performance was also found to be impaired in these animals, as compared to the control group. Our results suggest that the rat is a suitable model for brain dysfunction studies in Chagas' disease. PMID- 11403960 TI - Distinct effect of orphanin FQ in nucleus raphe magnus and nucleus reticularis gigantocellularis on the rat tail flick reflex. AB - The aim of the present study is to investigate the effects of orphanin FQ (OFQ) microinjected into the nucleus raphe magnus (NRM) and the nucleus reticularis gigantocellularis (NGC) on pain modulation. The tail-flick latency (TFL) was used as a behavioral index of nociceptive responsiveness. The result showed microinjection of OFQ into the NRM significantly increased the TFL, whereas microinjection of OFQ into the NGC decreased the TFL, suggesting the analgesic effect of OFQ in the NRM and the hyperalgesic effect of OFQ in the NGC. As there are three classes of putative pain modulating neurons in the rostral ventromedial medulla (RVM), the hyperalgesic or analgesic effect of OFQ in the RVM might depend upon the different class of the neurons being acted. PMID- 11403961 TI - Humans adapt the initial posture in learning a whole-body kicking movement. AB - What strategies are used in learning to control new movements? The present investigation sought to understand this process by analyzing the changes in whole body kinematics that occurred when subjects attempted to learn an unusual kicking movement. Five novices were taught a capoeira kick that involved both the upper and lower body for balance and co-ordination. Subjects performed two sets of 60 consecutive kicks, 24 h apart. Gradual changes in the body movement and the initial posture were found. Four subjects reduced the dynamic counter-twist associated with kick initiation. These subjects also adopted a more forward initial body lean. This gradual change in initial posture appeared to obviate the early counter-twist and to facilitate both the equilibrium and the goal directed components of the kick. PMID- 11403962 TI - Schwann cell apoptosis in experimental autoimmune neuritis of the Lewis rat and the functional role of tumor necrosis factor-alpha. AB - Schwann cell (SC) apoptosis may be a critical factor challenging nerve remyelination and regeneration in experimental autoimmune neuritis (EAN) in the Lewis rat. We therefore analyzed the fate of SC during high-dose antigen therapy of adoptive transfer-(AT-) EAN using rhP2 protein. P2 antigen therapy was associated with an increase of tumor necrosis factor (serum levels 1 h after intravenous (i.v.) injection and an augmentation of T-cell apoptosis. Antigen specific therapy had no clear effect on SC apoptosis. The effects on SC apoptosis were determined by morphological criteria or by in situ tailing (IST) followed by immunocytochemical analysis. Secondly, we neutralized TNF-alpha, released in abundance by antigen treatment but only in small concentrations during natural disease course. We found that the addition of a TNF-alpha neutralizing antiserum resulted in a significant decrease in the rate of SC apoptosis in vivo compared to animals treated with control antigen rhP0 or with rhP2 only. PMID- 11403963 TI - The N-methyl-D-aspartate receptor channel blockers memantine, MRZ 2/579 and other amino-alkyl-cyclohexanes antagonise 5-HT(3) receptor currents in cultured HEK-293 and N1E-115 cell systems in a non-competitive manner. AB - The type 3 serotonin (5-HT(3)) receptor is a ligand-gated ion channel. In concentration-clamp experiments, we investigated the effects of the uncompetitive N-methyl-D-aspartate (NMDA) receptor antagonists memantine, amantadine and MRZ 2/579 on 5-HT receptors stabley expressed in HEK-293 cells and on native 5-HT(3) receptors in the N1E-115 cell line. All agents antagonized serotonin (10 microM) induced inward currents with similar potency to that reported for NMDA receptors. This effect was characterized by inducing a pronounced receptor desensitization, and was probably non-competitive and voltage-independent. In contrast, (S) ketamine was much weaker as an antagonist of 5-HT(3) receptors than NMDA receptors. Similar effects on 5-HT(3) receptors have been reported previously for a variety of anti-depressants and it is possible that the clinical anti depressant effects reported for both memantine and amantadine are mediated, at least in part, by antagonistic effects at 5-HT(3) receptors. PMID- 11403964 TI - Cleaved caspase-3, caspase-7 and poly (ADP-ribose) polymerase are complementarily but differentially expressed in human medulloblastomas. AB - Expression of cleaved caspase-3, cleaved caspase-7 and specific product of caspase-dependent Poly (ADP-Ribose) Polymerase (PARP) cleavage have been examined by immunohistochemistry in seven human medulloblastomas. Cleaved caspase-3 and cleaved PARP expression parallels apoptosis as revealed with classical morphological criteria and with the method of in situ end-labelling of nuclear DNA fragmentation. Cleaved PARP co-localizes cleaved caspase-3 in the majority of tumors and areas thus indicating that caspase-3 is a major effector caspase leading to apoptosis in these tumors. Yet cleaved caspase-7 was also expressed in a small number of cells in four of seven tumors, but was the predominant caspase associated with cleaved PARP in one medulloblastoma. These findings indicate that effector caspase-3 and -7 may act in association, although caspase-7 may be exceptionally dominant in selected tumors. PMID- 11403965 TI - SHIRPA, a protocol for behavioral assessment: validation for longitudinal study of neurological dysfunction in mice. AB - Mouse models of neurological abnormalities are only valuable if accurately assessed. The three-stage SHIRPA procedure is used for the standardised assessment of mouse phenotype and has been reported in a high throughput experiment in which different mutants were ascertained at one age point using stage 1 of the protocol. In this study we have validated SHIRPA using a large cohort with one single mutation, 'legs at odd angles that causes neurological dysfunction. The cohort aged from 1 to 16 months during this study and this is the first longitudinal SHIRPA analysis. PMID- 11403966 TI - Acute and chronic haloperidol treatments increase parkin mRNA levels in the rat brain. AB - Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism. We examined the effects of acute and chronic treatment with haloperidol on parkin mRNA expression in the rat brain by reverse transcription-polymerase chain reaction. Acute haloperidol treatment (2 mg/kg) increased parkin mRNA levels in the striatum and nucleus accumbens but not in the medial prefrontal cortex and substantia nigra. Four-week-treatment with haloperidol decanoate (25 mg eq/kg) produced a significant increase in parkin mRNA levels in the striatum without affecting to those in the medial prefrontal cortex, nucleus accumbens and substantia nigra. These results suggest that Parkin may be involved in the haloperidol-induced synaptic plasticity, since Parkin regulates the turnover of the synaptic protein, CDCrel-1. PMID- 11403967 TI - Vigor of visceromotor responses to urinary bladder distension in rats increases with repeated trials and stimulus intensity. AB - This methodological study characterized visceromotor responses (VMRs) as abdominal contractile responses to urinary bladder distension (UBD) in the female rat. Electromyographic activity of the abdominal musculature was used as a measure of the VMR. Similar to previously characterized cardiovascular responses to UBD, VMRs to UBD demonstrated an initial sensitization period whereby repeated presentation of UBD stimuli led to increase vigor of the VMR. Graded UBD produced graded VMRs, therefore stimulus-response functions could be constructed. The intravenous administration of the opioid fentanyl produced a reduced vigor of the VMR in a fashion consistent with its analgesic effect. The present report supports the utility of this model for studies of urinary bladder nociception. PMID- 11403968 TI - A metabotropic glutamate receptor antagonist, alpha-methyl-4 carboxyphenylglycine, attenuates immediate early gene mRNA expression following traumatic injury in cultured rat cortical glial cells. AB - The effects of three glutamate receptor antagonists, (5R,10S)-(+)-5-methyl-10,11 dihydro-5H-dibenzo[a,d]-cyclohepten-5,10-imine hydrogen maleate (MK-801) for the N-methyl-D-aspartate receptor, 1,2,3,4-tetrahydro-6-nitro-2,3-dioxo-benzo[f] quinoxaline-7-sulfonamide (NBQX) for the alpha-amino-3-hydroxy-5methyl-4 isoxazole propionate /kinate receptor and (S)-alpha-methyl-4-carboxyphenylglycine (MCPG) for the metabotropic receptor, on c-fos and c-jun mRNA expression were investigated in cultured cortical glial cells following traumatic scratch injury. Expression of the two genes along the edges of wounds detected by in situ hybridization was not affected by MK-801 and NBQX. However, 100 and 500 microM of MCPG remarkably reduced the hybridization signals for both c-fos and c-jun mRNAs. The present results suggest that group I metabotropic glutamate receptors might have some association with immediate early gene induction after in vitro traumatic injury in glial cells. PMID- 11403969 TI - Temporal dynamics of the partial pressure of brain tissue oxygen during functional forepaw stimulation in rats. AB - The partial pressure of tissue oxygen (pO2) was measured in rat somatosensory cortex during periodic electrical forepaw stimulation of either 1 min or 4 s in duration, and correlated with simultaneous laser Doppler flowmetry. For both stimulus durations, a transient decrease in tissue pO2 preceded blood flow changes, followed by a peak in blood flow and an overshoot in tissue pO2. With protracted stimulation, tissue pO2 remained only slightly above pre-stimulus baseline, while blood flow was maintained at a reduced plateau phase. A sustained post-stimulus undershoot in tissue pO2 was observed only for the 1 min stimulus. These findings suggest a complex dynamic relationship between oxygen utilization and blood flow. PMID- 11403970 TI - Producing the vowel/a/ prior to speaking inhibits stuttering in adults in the English language. AB - This study investigated the effects of producing and listening to the vowel /a/ on the frequency of overt stuttering moments in eight people who stuttered. Stuttering frequency counts were made for the speech produced in the control condition, and after each of these four experimental conditions: (a) producing a vowel /a/ for 4 s; (b) producing a vowel /a/ for 4 s and waiting for 4 s; (c) listening to a recording of the vowel /a/ for 4 s; and (d) listening to a recording of the vowel /a/ for 4 s and waiting for 4 s. A significant reduction in the stuttering frequency was only observed following production of the vowel /a/ without a 4 s delay (P=0.02), suggesting that the vowel production prior to speech, serves as a temporary fluency enhancer. Its similarity to the occurrence of overt stuttering moments (e.g. discrete part-word repetitions and prolongation's) and its relationship to the fundamental nature of the pathology are discussed. PMID- 11403971 TI - Intraneuronal Abeta accumulation precedes plaque formation in beta-amyloid precursor protein and presenilin-1 double-transgenic mice. AB - beta-Amyloid peptides are key molecules that are involved in the pathology of Alzheimer's disease (AD). The source and place of the neurotoxic action of Abeta, however, is still a matter of controversial debates. In the present report, we studied the neuropathological events in a transgenic mouse model expressing human mutant beta-amyloid precursor protein and human mutant presenilin-1 in neurons. Western blot and immunohistochemical analysis revealed that intracellular Abeta staining preceded plaque deposition, which started in the hippocampal formation. At later stages, many neuritic Abeta positive plaques were found in all cortical, hippocampal and many other brain areas. Interestingly, intraneuronal Abeta staining was no longer detected in the brain of aged double-transgenic mice, which correlates with the typical neuropathology in the brain of chronic AD patients. PMID- 11403972 TI - Identification of differentially expressed genes in the denervated rat hippocampus by cDNA arrays. AB - To elucidate the molecular mechanism underlying the physiological responses to injury in the central nervous system, gene expression profiles in rodent hippocampus following perforant path transection were investigated using cDNA array hybridization. Of the 8000 arrayed clones, 47 exhibited differential expression by >3-fold difference in the denervated hippocampus from control, with 15 up-regulated and 22 down-regulated. They can be functionally assigned into several classes, among which the most prominent are those coding proteins involved in macromolecules synthesis and processing. Northern blot analysis verified the validation of the aforementioned array data. These results throw some new light on the physiological responses of the hippocampus to entorhinal deafferentation at molecular level. PMID- 11403973 TI - The role of preoptic area and anterior hypothalamus and median raphe nucleus on thermoregulatory system in freely moving rats. AB - To clarify the role of the preoptic area and anterior hypothalamus (PO/AH) on thermoregulatory system and the effects of serotonergic innervation from the median raphe nucleus (MRN) on body temperature (Tb), we perfused tetrodotoxin (TTX) solution into the PO/AH or MRN by using a microdialysis technique at different ambient temperatures (5, 23 and 35 degrees C) in freely moving rats. Tb was continuously monitored by using a telemetry system. In the MRN, perfusion of TTX solution induced significant hypothermia in the normal environment, a greater decrease in Tb during cold exposure and had no effect on Tb during heat exposure. In the PO/AH, perfusion of TTX solution induced significant hyperthermia in normal environment, a greater increase in Tb during heat exposure and had no effect on Tb during cold exposure. Our results indicate that the PO/AH regulates mainly heat loss or inhibits the loci regulating heat production. Furthermore, heat production appears to be regulated by other loci receiving serotonergic innervation from the MRN. PMID- 11403974 TI - Allele varepsilon4 of apolipoprotein E gene is less frequent in Down syndrome patient of the Sicilian population and has no influence on the grade of mental retardation. AB - We evaluated the allele (and genotype frequencies in 60 Down syndrome (DS), 25 mothers and 57 controls from Sicily and its relation with mental retardation. DS patients and sex ratio (M:F) was 22.1+/-10.5 and 1.14, respectively. Allele varepsilon4 and varepsilon3 frequencies were respectively lower (P=0.015) and higher (P=0.005) in DS patients compared to controls. Genotype varepsilon3/varepsilon4 and varepsilon3/varepsilon3 were less (P=0.03) and more frequent (P=0.001) in DS patients, with respective odd ratios of 0.31 (CI at 95%: 0.18-0.49) and of 4.4 (CI at 95%: 3.4-5.7). No difference of allele (distribution was found in function of the grades of mental retardation according to DMS-IV. Our results show that the implication of Apo-E4 in the pathogenesis of Alzheimer disease cannot be extrapolated in that of dementia of DS. PMID- 11403975 TI - Reduced gating of middle-latency auditory evoked potentials (P50) in migraine patients: another indication of abnormal sensory processing? AB - Habituation of cortical evoked responses to repetitive stimuli is reduced in migraine between attacks. To explore another aspect of information processing, we measured auditory sensory gating. The amplitude of the P50 response to the second of two homologous stimuli was significantly less reduced in migraineurs than in healthy volunteers. This lack of auditory sensory gating may be due to a hypofunction of monoaminergic subcortico-cortical pathways, which is also supposed to cause the interictal deficit of cortical habituation to repetitive stimuli. PMID- 11403976 TI - Effects of gonadal steroids on peripheral benzodiazepine receptor density in women with PMS and controls. AB - BACKGROUND: GABA receptor-modifying neurosteroids may play a role in premenstrual syndrome (PMS). The peripheral benzodiazepine receptor (PBR) both regulates the formation of neurosteroids and is, in animals, regulated by ovarian steroids. Alterations in PBR density have been observed in association with several psychiatric disorders. METHODS: We examined the effects of gonadal steroids on lymphocytic PBR density in nine women with prospectively confirmed PMS and nine controls. PBR densities were measured during three pharmacologically controlled conditions: gonadotropin releasing hormone agonist (Lupron)-induced hypogonadism, Lupron plus estradiol, and Lupron plus progesterone replacement. Blood samples were obtained after six weeks of Lupron alone and after 3-4 weeks of estradiol and progesterone replacement. RESULTS: No significant hormone state-related changes in PBR density were observed (ANOVA-R: phase-F(2,32)=1.5, P=0.2). Despite mood symptom development in the subjects with PMS, PBR density did not differ in women with PMS compared to controls across hormonal states (ANOVA-R: F(1,16)=0.6, P=0.4). CONCLUSIONS: PBR densities are not altered in women with PMS and are not changed significantly by selective gonadal steroid administration. Changes in PBR density would not appear to underlie the differential sensitivity to the mood destabilizing effects of ovarian steroids in PMS. PMID- 11403977 TI - Binding of [3H]paroxetine to serotonin uptake sites and of [3H]lysergic acid diethylamide to 5-HT2A receptors in platelets from women with premenstrual dysphoric disorder during gonadotropin releasing hormone treatment. AB - Changes in serotonergic parameters have been reported in psychiatric conditions such as depression but also in the premenstrual dysphoric disorder (PMDD). In addition, hormonal effects on serotonergic activity have been established. In the present study, binding of [3H]paroxetine to platelet serotonin uptake sites and binding of [3H]lysergic acid diethylamide ([3H]LSD) to platelet serotonin (5 HT)2A receptors were studied in patients with PMDD treated with a low dose of a gonadotropin releasing hormone (GnRH) agonist (buserelin) or placebo and compared to controls. The PMDD patients were relieved of premenstrual symptoms like depression and irritability during buserelin treatment. The number of [3H]paroxetine binding sites (Bmax) were significantly higher in the follicular phase in untreated PMDD patients compared to controls. When treated with buserelin the difference disappeared. No differences in [3H]LSD binding between the three groups were shown. The present study demonstrated altered platelet [3H]paroxetine binding characteristics in women with PMDD compared to controls. Furthermore, [3H]paroxetine binding was affected by PMDD treatment with a low dose of buserelin. The results are consistent with the hypothesis that changes in serotonergic transmission could be a trait in the premenstrual dysphoric disorder. PMID- 11403978 TI - Repeated morphine administration during pregnancy attenuates maternal behavior. AB - The present study tested the hypothesis that repeated administration of morphine on days 11-18 of pregnancy alters maternal behavior. Saline- and morphine-treated mothers were observed with their pups in two experiments. Rats were always tested twice a day during the light and dark phases of the reverse light/dark cycle. In Experiment 1, 12 types of activities and three types of nursing positions of mothers were recorded ten times during each 50-minute session for the 23-day lactation period. A decrease in nursing and active maternal behavior, and an increase in self-care, rearing and sniffing was found in morphine-treated mothers. Additionally, both saline- and morphine-treated mothers exhibited significantly more maternal behavior during the light, and non-maternal activities during the dark sessions of each day. Moreover, both saline- and morphine-treated mothers displayed significantly less maternal behavior and more non-maternal activities as postpartum time progressed. In Experiment 2, a different group of mothers was tested for pup retrieval from postnatal days 1 through 12. Morphine-treated mothers were slower than saline-treated mothers in retrieving all pups into the nest. However, there were no differences in latency to carry the first pup and return him/her to the nest. No unusual maternal behaviors were observed during the retrieval tests. Thus, the present study suggests that morphine administration during the second half of pregnancy attenuates some components of maternal behavior and increases non-maternal activities of mothers. PMID- 11403979 TI - Prefrontal cortex as the site of estrogen's effect on cognition. AB - The hippocampus has long been presumed the primary site of action of estrogens on cognition; and explicit memory is considered the cognitive function most vulnerable to menopausal loss of estrogen. We hypothesize instead that the prefrontal cortex and its neural circuitry are prime mediators of estrogen's role in cognition. We also propose that previously reported menopausal cognitive decline, presumed to be hippocampally mediated, may be secondary to executive dysfunction. We used a cross sectional design to compare the performance of nine menopausal women on hormone replacement therapy (HRT) and 10 menopausal women with no prior exposure to HRT on a battery of neuropsychological tests. The battery was comprised primarily of tests of memory and executive functioning. Executive functioning is mediated by the frontal lobes and encompasses working memory, directed attention, the inhibition of inappropriate responses, cognitive set switching, and behavioral monitoring. Unlike most previous studies, we used a memory measure that yields multiple scores reflecting various problem-solving strategies and error types, thus isolating spared and impaired cognitive processes. Results yielded both qualitative and quantitative evidence for disruption of cognitive processes subserved by the frontal lobes rather than the hippocampus: 1) despite intact free recall on a list-learning task (CVLT), untreated menopausal women were relatively impaired in correctly recognizing words previously learned and distinguishing them from items not on the list (discriminability), 2) untreated women also had difficulty inhibiting inappropriate responses in the form of perseverative errors, and 3) the non-HRT group consistently performed worse on the N-back test of working memory. The prefrontal cortex is critical for intact working memory and estrogen enhances performance on working memory tasks. In conclusion, this study provides preliminary evidence for executive dysfunction in untreated menopausal women as women with HRT outperformed women without HRT on tests requiring directed attention, inhibition of inappropriate responses, and cognitive set switching. PMID- 11403980 TI - Salivary cortisol and DHEA: association with measures of cognition and well-being in normal older men, and effects of three months of DHEA supplementation. AB - Dehydroepiandrosterone (DHEA) is a steroid that shows a marked age-related decline in humans. Previous research suggests potential for DHEA replacement in old age to enhance cognition and well-being. We conducted a clinical trial to test these hypotheses in a non-clinical sample of 46 men aged 62-76. Participants received either 50 mg DHEA daily for 13 weeks, followed by placebo for 13 weeks, or the reverse, in a randomised double-blind cross-over trial design. Levels of salivary cortisol and DHEA were measured at 0800 h and 2000 h prior to each assessment session. Cognition was assessed with tests of speed, attention and episodic memory. Well-being was measured with questionnaires of mood and perceived health. Mood questionnaires were completed at the assessment session as well as concurrently with saliva sampling.A correlational analysis of baseline behavioural data with hormonal data, controlling for age, revealed that higher morning DHEA was associated with lower confusion (r=-0.33; P=0.04), while higher evening DHEA was associated with lower anxiety (r=-0.35; P=0.03) and lower current negative mood in the morning (r=-0.37; P=0.03). Conversely, higher morning cortisol and a morning cortisol/DHEA ratio were associated with higher anxiety (r=0.35; P=0.03), (r=0.46; P=0.004), general mood disturbance (r=0.32; P=0.046), (r=0.32; P=0.04) and higher current negative mood in the evening (r=0.37; P=0.03), (r=0.38; P=0.03). A higher morning cortisol/DHEA ratio was also associated with higher confusion (r=0.39; P=0.01) and lower visuo-spatial memory performance (r=-0.39; P=0.01). Unexpectedly, higher evening cortisol was associated with faster choice reaction time (r=-0.33; P=0.04). These findings are consistent with an impairing effect of high cortisol on episodic memory and mood in older men, which may be attenuated by DHEA. When treatment effects were analysed, no significant effects of DHEA were observed on any of the trial outcomes, providing no support for benefits of DHEA supplementation for cognition or well-being in normal older men in the shorter-term. PMID- 11403981 TI - Association between time of awakening and diurnal cortisol secretory activity. AB - A 12-hour diurnal profile of salivary free cortisol was measured in healthy adults (n=40) on two consecutive days. Samples were collected at timed intervals synchronised to awakening. The mean profile is characterised by a marked increase in cortisol concentration following awakening, peaking after about 30 min, and a subsequent decline over the remainder of the day. Thus two components of the diurnal cycle were examined: a) the first 45 min post-awakening (the awakening cortisol response) and b) the underlying 12 h profile from immediately until 12 h post awakening (but without the awakening response). Both of these components were analysed in two ways such as to provide an indication of overall cortisol concentration and the degree of change in cortisol concentration, i.e. the rise for the awakening response and the diurnal decline. Both components of the cortisol diurnal profile were negatively correlated with awakening time. Thus, those subjects who awoke earliest had higher levels of cortisol over the 45 min following awakening as well as throughout the rest of the day. They also displayed a more marked diurnal decline to be convergent with late awakeners at the end of diurnal measurement, 12 h following awakening. Hence the diurnal cortisol cycle, which is synchronised to awakening, is significantly related to awakening time. These findings support the notion of a close association between suprachiasmatic nucleus (SCN) control of both awakening and cortisol secretory activity. PMID- 11403982 TI - Fluctuations in spatial recognition memory across the menstrual cycle in female rhesus monkeys. AB - Findings are inconsistent regarding whether women's cognitive performance fluctuates across phases of the menstrual cycle, but differences in methodology and the use of reported cycle phase rather than precise hormonal measures may underlie these disparities. Studies in monkeys may help resolve these discrepant findings, since hormonal status can be reliably determined. We tested four young (5-7 years old) female rhesus monkeys daily during one entire menstrual cycle on three cognitive tasks displayed on a computerized touch-screen system: a Matching to Sample task with a 30 s delay (MTS-30s), a Matching to Sample task without delay (MTS-no delay) and the spatial condition of the Delayed Recognition Span Test (spatial-DRST). Blood samples were collected at specific time intervals throughout the cycle and assayed for estradiol and progesterone in order to identify hormonal status. There was a nonsignificant trend for the MTS-30s scores to be better during the follicular and luteal phases, when estradiol levels were low, than during the peri-ovulatory phase, when estradiol levels were at their highest. MTS-no delay performance did not vary as a function of hormonal status. Spatial-DRST scores were significantly better during the follicular and luteal phases than during the peri-ovulatory phase of the cycle. These data in the female rhesus monkey support the hypothesis that spatial memory performance is sensitive to estradiol variations across the menstrual cycle, with better performance associated with low estradiol levels. PMID- 11403983 TI - Effects of olanzapine and haloperidol on serum prolactin levels in male schizophrenic patients. AB - It has been proposed that new atypical antipsychotics cause minimal prolactin (PRL) elevation compared to traditional antipsychotic agents because they spare dopamine blockade within the brain's tuberoinfundibular tract. The aim of this study was to compare the effects of olanzapine and haloperidol on PRL secretion in male schizophrenic patients. Twenty-nine male schizophrenic inpatients were included in the study. Fifteen of them were given olanzapine in a fixed dose of 10 mg/day PO and 14 of them were given haloperidol in a fixed dose of 10 mg/day PO for 6 weeks after a 2-week drug washout period. Fifteen age-matched healthy control subjects were used as control group. PRL levels were measured both before and after the 6-week treatment period in the patients. At the end of the 6th week, the PRL values observed with olanzapine treatment were significantly less than those observed with haloperidol, but not different from those of the controls. There was a significant positive correlation between the PRL values and the severity of extrapyramidal side effects in only the haloperidol group after the six week's treatment period. Our data indicate that short-term olanzapine treatment at doses of 10 mg/day PO causes minimal elevations in PRL secretion in male schizophrenic patients in contrast to haloperidol. This finding is consistent with the previous reports and may be attributed to olanzapine's differential effects on dopamine neurotransmission. PMID- 11403984 TI - The mammalian pituitary intermediate lobe: an update on innervation and regulation. AB - The pituitary intermediate lobe (IL) in mammals is an area of uniform endocrine cells which synthesize and release specific peptide products of the proopiomelanocortin gene. The lobe receives direct synaptic connections onto the endocrine cells from hypothalamic dopaminergic neurons. This review updates information on the dopaminergic as well as the gamma-aminobutyric acid inhibitory neuroregulation for the IL. It also provides a discussion of stimulatory molecules which are likely to affect peptide release, particularly the neurotransmitter serotonin, which may be present via uptake into the dopaminergic nerve terminals. Other stimulatory molecules discussed which are likely to significantly affect peptide secretion are norepinephrine, corticotropin releasing factor, and several opiate peptides. A new direction of study involves the potential interaction of neurotrophic factors, which are present in all areas of the pituitary, and may be suggested to have a supportive role for the neural elements of the IL. The endocrine cells of the IL and their direct hypothalamic innervation are considered to be an easily accessed peripheral model for study of both neural-endocrine and neurotrophic-target cell interactions. PMID- 11403985 TI - Role of the alpha(1)- and alpha(2)-adrenoceptors of the paraventricular nucleus on the water and salt intake, renal excretion, and arterial pressure induced by angiotensin II injection into the medial septal area. AB - In this study we investigated the influence of alpha-adrenergic antagonists injections into the paraventricular nucleus (PVN) of the hypothalamus on the thirst and salt appetite, diuresis, natriuresis, and pressor effects of angiotensin II (ANG II) stimulation of medial septal area (MSA). ANG II injection into the MSA induced water and sodium intake, diuresis, natriuresis, and pressor responses. The previous injection of prazosin (an alpha(1)-adrenergic antagonist) into the PVN abolished, whereas previous administration of yohimbine (an alpha(2) adrenergic antagonist) into the PVN increased the water and sodium intake, urinary, natriuretic, and pressor responses induced by ANG II injected into the MSA. Previous injection of a nonselective alpha-adrenergic antagonist, regitin, into the PVN blocked the urinary excretion, and reduced the water and sodium intake, sodium intake, and pressor responses induced by ANG II injected into the MSA. The present results suggest that alpha-adrenergic pathways involving the PVN are important for the water and sodium excretion, urine and sodium excretion, and pressor responses, induced by angiotensinergic activation of the MSA. PMID- 11403986 TI - Decreased striatal dopamine efflux after intrastriatal application of benzazepine class D1 agonists is not mediated via dopamine receptors. AB - Previous pharmacological studies have reported that striatal dopamine efflux is negatively modulated not only by presynaptic D2 dopamine autoreceptors but also by striatal D1 dopamine receptors. The present experiments employed in vivo microdialysis to further examine the ability of widely used benzazepine-class D1 agonists to modulate striatal dopamine efflux. In the present study, both the partial D1 agonist (+/-)-SKF 38393 (10 microM) and the full D1 agonist (+/-)-SKF 82958 (10 and 100 microM) significantly reduced striatal dopamine efflux during intrastriatal application. Intrastriatal application of the less active enantiomer, S(-)-SKF 38393 (10 microM) did not decrease striatal dopamine suggesting a selective receptor-mediated mode of action of (+/-)-SKF 38393. Additional experiments were conducted with the full D1 agonist (+/-)-SKF 82958 in order to characterize the receptor(s) mediating the observed decrease in dopamine efflux. Neither local application of the D1 antagonist R(+)-SCH 23390 (100 microM) nor local application of the selective D2 antagonist raclopride (5 microM) blocked the ability of (+/-)-SKF 82958 (10 microM) to decrease striatal dopamine efflux. However, intrastriatal application of the less selective D2 antagonist haloperidol (1 microM) did prevent the decrease in striatal dopamine efflux observed during intrastriatal (+/-)-SKF 82958 application. The present data suggest that the ability of intrastriatally applied benzazepine-class D1 agonists to decrease striatal dopamine efflux is receptor-mediated, but this action apparently is not mediated at D1 or D2 receptors. There is therefore no indication for an intrastriatal population of D1 receptors capable of modulating dopamine efflux. PMID- 11403987 TI - Coriolis-induced cutaneous blood flow increase in the forearm and calf. AB - Using venous occlusion plethysmography, Sunahara et al. reported that Coriolis induced nausea was accompanied by an increase in forearm blood flow, suggesting a decrease in sympathetic activity to this vascular bed. No significant blood pressure and heart rate changes were observed. Vasodilation of the limbs theoretically impairs orthostatic tolerance, particularly if blood flow is shown to increase simultaneously in the lower limbs. This study examined the latter possibility. Seventeen subjects were exposed to the Coriolis cross-coupling effects induced by 20 RPM yaw rotation, and a simultaneous 45 degrees pitch forward head movement in the sagittal plane every 12 s. Forearm and calf skin blood flow were monitored in real-time using laser Doppler flowmetry (PeriFlux 4001). Our results indicated a significant (p < 0.001) simultaneous forearm and calf skin blood flow increase as a result of Coriolis cross-coupling across all 15 susceptible subjects. No significant changes in blood pressure and heart rate were observed. Coriolis-induced cardiovascular changes may confound previous reports on reduced G tolerance using ground-based centrifuges that invariably evoke cross-coupling effects. PMID- 11403988 TI - Collateral projections from the median raphe nucleus to the medial septum and hippocampus. AB - It has previously been shown that the median raphe nucleus (MR) is a source of pronounced projections to the septum and hippocampus. The present study examined collateral projections from MR to the medial septum (MS) and to various regions of the hippocampus. The fluorescent retrograde tracers, Fluororuby and Fluorogold, were injected into the septum and hippocampus, respectively, and the median raphe nucleus was examined for the presence of single- and double-labeled neurons. The dorsal raphe nucleus (DR) was also examined for the presence of single- and double-labeled cells and comparisons were made with the MR. The main findings were: (1) pronounced numbers of retrogradely labeled cells (approximately 50 cells/section) were present in MR with injections in the MS or in various regions of the hippocampus; (2) approximately 8-12% of MR cells were double-labeled following paired injections in the MS-CA1, MS-CA3, and MS-dentate gyrus of the dorsal hippocampus, the lateral MS-dentate gyrus, and the MS-ventral hippocampus; (3) single- and double-labeled cells were intermingled throughout MR and present in greater numbers in the rostral than caudal MR; and (4) significantly more single- and double-labeled cells were present in MR than in DR with all combinations of injections. These findings demonstrate that MR projects strongly to the MS and hippocampus, and that a significant population of MR neurons (8-12%) sends collateral projections to both sites. It is well established that the MR nucleus serves a direct role in the desynchronization of the electroencephalographic (EEG) activity of the hippocampus-or the blockade of the hippocampal theta rhythm. The MR neurons that we have identified with collateral projections to the septum and hippocampus may be critically involved in the modulation/control of the hippocampal EEG. A role for the MR in memory associated functions of the hippocampus is discussed. PMID- 11403989 TI - Alteration of dopamine transport in the striatum and nucleus accumbens of ovariectomized and estrogen-primed rats following N-(p-isothiocyanatophenethyl) spiperone (NIPS) treatment. AB - The ability of N-(p-isothiocyanatophenethyl) spiperone (NIPS, 10 mg/kg, 24 h), a selective, irreversible alkylating agent of the dopamine D(2) receptor, to alter properties of dopamine uptake and clearance in the striatum and nucleus accumbens of ovariectomized and estrogen-primed (estradiol benzoate, 10 microg, 48 h, 24 h) rats was examined using voltammetry. The effectiveness of NIPS was evaluated independently by measuring agonist mediated potentiation of [35S]-guanosine 5' (gamma-thiotriphosphate) ([35S]-GTPgammaS) binding and [3H]-dopamine uptake. A decrease in E(max) for ligand potentiated [35S]-GTPgammaS binding and a loss of quinpirole potentiated [3H]-dopamine uptake was observed consistent with a NIPS mediated alkylation and functional down-regulation of the dopamine D(2) receptor. This down-regulation was associated with an attenuation of the dose dependent uptake of dopamine in both the striatum and the accumbens. Co-administration of estrogen and NIPS resulted in a further attenuation of dopamine potentiated [35S] GTPgammaS binding measured in vitro and dopamine uptake measured in vivo. Analysis of the voltammetric profile revealed that clearance and T(50) times were significantly prolonged in animals treated with estrogen and NIPS compared with those treated with NIPS alone. These data are consistent with both a steroid mediated impairment in dopamine autoreceptor/dopamine transporter coupling and an independent action of estrogen at the level of the dopamine transporter. PMID- 11403990 TI - Cholinergic, noradrenergic, and serotonergic inhibition of fast synaptic transmission in spinal lumbar dorsal horn of rat. AB - It is known that spinal nociceptive sensory transmission receives descending inhibitory and facilitatory modulation from supraspinal structures. Glutamate is the major fast excitatory transmitter between primary afferent fibers and spinal dorsal horn neurons. In whole-cell patch clamp recordings from dorsal horn neurons in spinal slices, we investigated synaptic mechanisms for inhibitory modulation at the lumbar level of the spinal cord. Application of the cholinergic receptor agonist carbachol produced a dose-dependent inhibition of glutamate mediated excitatory postsynaptic currents (EPSCs) (IC(50) 13 microM). Postsynaptic injection of two different types of G-protein inhibitors, guanosine 5'-O-2-thiophosphate or guanosine 5'-O-3-thiotriphosphate, blocked the inhibition produced by carbachol. Clonidine, a selective alpha-adrenergic receptor agonist, also produced a dose-dependent inhibition of EPSCs (IC(50) 7 microM) that was reduced by postsynaptic inhibition of G-proteins. The inhibitory effect of serotonin was likewise mediated by postsynaptic G-proteins. Our results suggest that activation of postsynaptic neurotransmitter receptors plays a critical role in inhibition of glutamate mediated sensory responses by acetylcholine, norepinephrine, and serotonin. Our results support the hypothesis that descending sensory modulation may be mediated by multiple neurotransmitter receptors in the spinal cord. PMID- 11403991 TI - Repeated neonatal maternal separation alters morphine-induced antinociception in male rats. AB - Recent studies indicate that Long-Evans rats separated from their dam for 3 h daily over the first 2 weeks of life (maternally separated [MS] rats) exhibit exaggerated behavioral and neuroendocrine responses to stress as adults compared to handled (H) or non-handled (NH) control animals. Our aim was to determine whether repeated neonatal maternal separation results in altered sensitivity to the opioid agonist morphine in male and female adult rats. Sensitivity to morphine was assessed using hot-plate and tail-flick tests. Morphine was less potent inducing antinociception in MS males compared to same-sex controls in the hot-plate, but not in the tail-flick test. Decrease in sensitivity to morphine in MS females compared to same-sex controls was present only as a trend in the hot plate, but not in the tail-flick test. These results suggest that neonatal maternal separation results in long-lasting changes in opioid responsiveness primarily in male rats. PMID- 11403992 TI - Orphanin FQ/nociceptin binds to functionally coupled ORL1 receptors on human immune cell lines and alters peripheral blood mononuclear cell proliferation. AB - Orphanin FQ/nociceptin (OFQ/N) has been shown to modulate nociception, responses to stress and anxiety. We investigated OFQ/N function in human immune cells. We find that monocytic U937, T lymphocytic CEM, and MOLT-4 cell lines express OFQ/N binding sites at levels comparable to that of human SH-SY5Y neuroblastoma cells. We show that OFQ/N receptors are functionally coupled to G proteins in these cells. Finally OFQ/N decreases proliferation of phytohemagglutinin-stimulated peripheral blood mononuclear cells in vitro at doses ranging from 10(-13) to 10( 8) M. Thus, our data suggest that OFQ/N and OFQ/N receptor may act as an immunomodulatory system. PMID- 11403993 TI - Membrane fluidity effects of estratrienes. AB - Estrogens have demonstrable neuroprotective effects. This fact has lead to the proposed use of estrogens for the prevention and/or treatment of Alzheimer's disease. The exact protective mechanism estrogens provide is not fully understood. In this report, a potential non-genomic mechanism for estratrienes involving alterations in membrane fluidity was studied. Steroids, such as estrogen, are known to be membrane-active and can alter the lipid packing. In this study we used fluorescent methodologies to address the effect of naturally occurring steroids (17alpha and 17beta-estradiol, testosterone, and progesterone) and new estratriene analogs on membrane fluidity using liposomes and HT-22 hippocampal cells. The study's results indicate steroids, based on the estratriene nucleus, can modulate lipid packing as evidenced by (1) decreased membrane fusion events and (2) decreased membrane fluidity. The effects on the membrane were both time and concentration dependent. It was also demonstrated through rational design estratriene analogs can be synthesized with enhanced membrane effects. Finally, in a glutamate-induced toxicity HT-22 model, we also demonstrated cellular protection with the estratriene-based molecules and analogs. The data suggest the plethora of cellular actions of estrogens may relate to or be influenced by membrane effects of the steroid. PMID- 11403994 TI - Enzyme histochemical profile of immunohistochemically identified Renshaw cells in rat lumbar spinal cord. AB - Activity levels of cytochrome oxidase, acid phosphatase, and NADPH diaphorase were examined in the perikarya of immunohistochemically identified Renshaw cells from sections of rat lumbar spinal cord. Renshaw cell profiles were identified on the basis of their characteristic anti-gephyrin-immunofluorescent labelling. Intrasomatic densities of enzyme histochemical reaction product were employed as indicators of relative mitochondrial activity (cytochrome oxidase), intracytoplasmic digestion (acid phosphatase), or putative nitrergic signalling (NAPDH-diaphorase). Approximately half of the Renshaw cell somata examined displayed moderate levels of cytochrome oxidase reaction product (142 of 262 Renshaw cells) or low levels of acid phosphatase activity (156 of 243 Renshaw cells). A majority (160 of 202 cells) of Renshaw cells contained low intrasomatic levels of NADPH-diaphorase activity but most of these cells were closely apposed by at least one NADPH-diaphorase reactive axonal varicosity. Our findings suggest that moderate levels of perikaryal oxidative metabolism and low levels of intracytoplasmic digestion are sufficient for, and support, the unique physiological capabilities of Renshaw cells. The presence of NADPH-diaphorase containing somatic close contacts indicate that nitric oxide may have at least a minor role in the regulation of Renshaw cell activity. These results are complementary and consistent with previous morphological and pharmacological demonstrations of Renshaw cell heterogeneity. PMID- 11403995 TI - Deprenyl stimulates the efflux of monoamines from the rat hypothalamus in vitro. AB - The direct effects of L-deprenyl, a monoamine oxidase inhibitor, on the hypothalamus of male Sprague-Dawley rats was investigated by measuring the efflux of norepinephrine (NE), dopamine (DA), serotonin (5-HT), dihydroxyphenylacetic acid (DOPAC), and 5-hydroxyindoleacetic acid (5-HIAA) using a combination of high performance liquid chromatography with electrochemical detection and an in vitro incubation system. After measuring basal efflux by incubating the hypothalami with Krebs-Ringers Henseleit (KRH) alone during the first incubation period, hypothalami were incubated either with the medium, KRH alone (0 mM), or KRH containing 0.1, 1, and 10 mM L-deprenyl. During the third incubation period, hypothalami were again incubated with KRH alone to measure the residual effects if any. During the final incubation period, the hypothalami were stimulated with high K(+) KRH. Deprenyl produced a dose-dependent increase in the efflux of NE, DA, and 5-HT from the hypothalami. Neurotransmitter efflux returned to pretreatment levels when L-deprenyl was removed from the medium. In contrast to NE, DA, and 5-HT, the efflux of the metabolites DOPAC and 5-HIAA was inhibited in a dose-dependent fashion after incubation with L-deprenyl. Results from this study demonstrate that L-deprenyl is capable of stimulating the efflux of neurotransmitters in vitro by a direct action on the hypothalamus. PMID- 11403996 TI - Male-female differences in rat hypothalamic-pituitary-adrenal axis responses to nicotine stimulation. AB - Hypothalamic-pituitary-adrenal (HPA) axis responsiveness differs physiologically and pharmacologically between the sexes (sexual diergism). Central nicotinic receptors modulate this endocrine axis. Previous studies have established that nicotine (NIC) stimulates the HPA axis; however, only male animals have been used. We have demonstrated that plasma arginine vasopressin (AVP) and adrenocorticotropic hormone (ACTH) concentrations showed greater responsiveness in male than in female rats pretreated with scopolamine (SCOP), a muscarinic antagonist, followed by physostigmine (PHYSO), an acetylcholinesterase inhibitor. These results suggest that the SCOP + PHYSO effects may have resulted from an indirect nicotinic effect caused by increased synaptic acetylcholine with simultaneous muscarinic antagonism. In the present study, we investigated nicotinic cholinergic influences on HPA axis activity in male and female rats by administering NIC (0, 0.03, 0.1, 0.3, or 0.5 mg/kg) and determining plasma AVP, ACTH, and corticosterone (CORT) responses. Male rats had a significantly greater, dose-related AVP response to NIC than did females. In contrast, female rats had significantly greater, dose-related ACTH and CORT responses to NIC than did males. Hormone responses following NIC were similar to hormone responses following SCOP + PHYSO. These results suggest nicotinic receptors influence the HPA axis differentially in male and female rats. PMID- 11403997 TI - The differential effect of angiotensin II and angiotensin 1-7 on norepinephrine, epinephrine, and dopamine concentrations in rat hypothalamus: the involvement of angiotensin receptors. AB - Angiotensin 1-7 has been recently claimed the active member of the angiotensins' family. In the present study we compared the effect of angiotensin II and angiotensin 1-7 on the concentration of dopamine, serotonin, epinephrine, and norepinephrine and some of their metabolites in the rat hypothalamus, where the levels of angiotensins are particularly high. Intracerebroventricular injection of angiotensin II, but not angiotensin 1-7, time-dependently elevated the levels of both epinephrine (p < 0.05) and norepinephrine (p < 0.05) in the hypothalamus and both effects could be prevented by intracerebroventricular injection of either AT(1) (candesartan), AT(2) (PD123319) or AT(1-7) (A-779) receptor antagonist. Neither angiotensin II nor angiotensin 1-7 produced any changes in the level of dopamine, dihydroxyphenylacetic acid, homovanilic acid, serotonin, 5 hydroxyindoleacetic acid, or tryptophan at any time point in comparison with the control groups. However, AT(1) but not AT(2) receptor blockade, unmasked the stimulatory effect of angiotensin 1-7 on dopamine concentration in the hypothalamus. Thus, angiotensin II and its active metabolite angiotensin 1-7 regulate selectively, albeit differentially, adrenergic, noradrenergic and dopaminergic systems in the hypothalamus, the effects that involve AT(1), AT(2) and AT(1-7) angiotensin receptors. PMID- 11403998 TI - Role of the multifunctional CDP/Cut/Cux homeodomain transcription factor in regulating differentiation, cell growth and development. AB - CDP/Cux/Cut proteins are an evolutionarily conserved family of proteins containing several DNA binding domains: one Cut homeodomain and one, two or three Cut repeats. In Drosophila melanogaster, genetic studies indicated that Cut functions as a determinant of cell-type specification in several tissues, notably in the peripheral nervous system, the wing margin and the Malpighian tubule. Moreover, Cut was found to be a target and an effector of the Notch signaling pathway. In vertebrates, the same functions appear to be fulfilled by two cut related genes with distinct patterns of expression. Cloning of the cDNA for the CCAAT-displacement protein (CDP) revealed that it was the human homologue of Drosophila Cut. CDP was later found be the DNA binding protein of the previously characterized histone nuclear factor D (HiNF-D). CDP and its mouse counterpart, Cux, were also reported to interact with regulatory elements from a large number of genes, including matrix attachment regions (MARs). CDP/Cut proteins were found generally to function as transcriptional repressors, although a participation in transcriptional activation is suggested by some data. Repression by CDP/Cut involves competition for binding site occupancy and active repression via the recruitment of a histone deacetylase activity. Various combinations of Cut repeats and the Cut homeodomains can generate distinct DNA binding activities. These activities are elevated in proliferating cells and decrease during terminal differentiation. One activity, involving the Cut homeodomain, is upregulated in S phase. CDP/Cut function is regulated by several post-translational modification events including phosphorylation, dephosphorylation, and acetylation. The CUTL1 gene in human was mapped to 7q22, a chromosomal region that is frequently rearranged in various cancers. PMID- 11403999 TI - Multiple alignment of complete sequences (MACS) in the post-genomic era. AB - Multiple alignment, since its introduction in the early seventies, has become a cornerstone of modern molecular biology. It has traditionally been used to deduce structure / function by homology, to detect conserved motifs and in phylogenetic studies. There has recently been some renewed interest in the development of multiple alignment techniques, with current opinion moving away from a single all encompassing algorithm to iterative and / or co-operative strategies. The exploitation of multiple alignments in genome annotation projects represents a qualitative leap in the functional analysis process, opening the way to the study of the co-evolution of validated sets of proteins and to reliable phylogenomic analysis. However, the alignment of the highly complex proteins detected by today's advanced database search methods is a daunting task. In addition, with the explosion of the sequence databases and with the establishment of numerous specialized biological databases, multiple alignment programs must evolve if they are to successfully rise to the new challenges of the post-genomic era. The way forward is clearly an integrated system bringing together sequence data, knowledge-based systems and prediction methods with their inherent unreliability. The incorporation of such heterogeneous, often non-consistent, data will require major changes to the fundamental alignment algorithms used to date. Such an integrated multiple alignment system will provide an ideal workbench for the validation, propagation and presentation of this information in a format that is concise, clear and intuitive. PMID- 11404000 TI - Mouse fatty acid transport protein 4 (FATP4): characterization of the gene and functional assessment as a very long chain acyl-CoA synthetase. AB - FATP4 (SLC27A4) is a member of the fatty acid transport protein (FATP) family, a group of evolutionarily conserved proteins that are involved in cellular uptake and metabolism of long and very long chain fatty acids. We cloned and characterized the murine FATP4 gene and its cDNA. From database analysis we identified the human FATP4 genomic sequence. The FATP4 gene was assigned to mouse chromosome 2 band B, syntenic to the region 9q34 encompassing the human gene. The open reading frame was determined to be 1929 bp in length, encoding a polypeptide of 643 amino acids. Within the coding region, the exon-intron structures of the murine FATP4 gene and its human counterpart are identical, revealing a high similarity to the FATP1 gene. The overall amino acid identity between the deduced murine and human FATP4 polypeptides is 92.2%, and between the murine FATP1 and FATP4 polypeptides is 60.3%. Northern analysis showed that FATP4 mRNA was expressed most abundantly in small intestine, brain, kidney, liver, skin and heart. Transfection of FATP4 cDNA into COS1 cells resulted in a 2-fold increase in palmitoyl-CoA synthetase (C16:0) and a 5-fold increase in lignoceroyl-CoA synthetase (C24:0) activity from membrane extracts, indicating that the FATP4 gene encodes an acyl-CoA synthetase with substrate specificity biased towards very long chain fatty acids. PMID- 11404001 TI - The chorion genes of the medfly. II. DNA sequence evolution of the autosomal chorion genes s18, s15, s19 and s16 in Diptera. AB - We present a total of approximately 15 kb of DNA sequences, encompassing four chorion genes Ccs18, Ccs15, Ccs19, Cc16 and their flanking DNA in the medfly C. capitata. Comparison of coding regions, introns and intergenic sequences in five Dipteran species, D. melanogaster, D. subobscura, D. virilis, D. grimshawi and C. capitata documented an extensive divergence in introns and coding regions, but few well conserved elements in the proximal 5' flanking regions in all species. These elements are related to conserved regulatory features of three of the genes, including tissue- and temporal regulation. In the fourth, gene s15, significant alterations in the 5' flanking region may be responsible for its changed temporal regulation in C. capitata. One long intergenic sequence, located in the distal 5' flanking region of gene s18, is homologous to ACE3, a major amplification control element and contains an 80-bp A/T-rich sequence, known to stimulate strong binding of the origin recognition complex (ORC) in D. melanogaster. Analysis of the nucleotide composition of all chorion genes in C. capitata and D. melanogaster showed that C. capitata exhibit less biased representation of synonymous codons than does D. melanogaster. PMID- 11404002 TI - Identification and localization of two mouse phosphomannomutase genes, Pmm1 and Pmm2. AB - Phosphomannomutases catalyze the reversible conversion of mannose 6-phosphate to mannose 1-phosphate. In humans, two different isozymes have recently been identified, PMM1 and PMM2. We have previously shown that mutations in the PMM2 gene cause the most frequent type of the congenital disorders of glycosylation, CDG-Ia. Here, we present data on the two mouse orthologous genes, Pmm1 and Pmm2. The chromosomal localization of the two mouse genes has been determined. We also present the gene structure and the exon-intron organization of Pmm1 and Pmm2. Pmm1 maps to mouse chromosome 15, Pmm2 to chromosome 16. These chromosomal regions are syntenic with regions on human chromosomes 22 and 16, respectively. The Pmm1 gene is composed of eight exons and spans approximately 9.5 kb. The genomic structure is extremely well conserved between the human and mouse gene. The Pmm2 gene consists of eight exons and spans a larger genomic region ( approximately 20 kb). An alignment of the human and mouse protein sequences confirms the conservation among this family of phosphomannomutases. The two mouse genes are expressed in many tissues, but the expression pattern is slightly different between Pmm1 and Pmm2. The most striking difference is the high expression of Pmm1 in brain tissue, whereas Pmm2 is only weakly expressed in this tissue. PMID- 11404003 TI - Transfer of high copy number plasmid into mammalian cells by calcium phosphate transfection. AB - Using flow cytometry, single cell sorting, confocal microscopy and fluorescent plasmids, a thorough study of DNA uptake, DNA fate and DNA expression in mammalian cells transfected with the widely used calcium-phosphate precipitation method was executed. We show for the first time that up to 100,000 plasmid molecules can be delivered into individual cells, but also that DNA transfer into cells is a dynamic process that follows a defined kinetics of uptake and intracellular processing. Analyses by flow cytometry and confocal microscopy have also supported results suggesting endocytosis during Ca-Pi transfection. We also demonstrate that expression-enhancing treatment with glycerol during transfection did not result in increased DNA uptake. While cells with maximal DNA load appear to express the highest level of the transgene, these cells are negatively impacted in terms of growth and survival. PMID- 11404004 TI - RAI1 is a novel polyglutamine encoding gene that is deleted in Smith-Magenis syndrome patients. AB - The human chromosomal band 17p11.2 is a genetically unstable interval. It has been shown to be deleted in patients suffering from Smith-Magenis syndrome. Previous efforts of physical and transcriptional mapping in 17p11.2 and subsequent genomic sequencing of the candidate interval allowed the identification of new genes that might be responsible for the Smith-Magenis syndrome. In this report, one of these genes named RAI1, the human homologue of the mouse Rai1 gene, has been investigated for its contribution to the syndrome. Expression analysis on different human adult and fetal tissues has shown the existence of at least three splice variants. Moreover, the most interesting feature of the gene is the presence of a polymorphic CAG repeat coding for a polyglutamine stretch in the amino terminal domain of the protein. PMID- 11404005 TI - Cloning of the bone Gla protein gene from the teleost fish Sparus aurata. Evidence for overall conservation in gene organization and bone-specific expression from fish to man. AB - Bone Gla protein (BGP, Osteocalcin) is a bone-specific vitamin K-dependent protein which has been intensively studied in mammals. Although BGP is the most abundant non-collagenous protein of bone, its mode of action at the molecular level remains unclear. From an evolutionary point of view, the appearance of BGP seems to parallel the appearance of hydroxyapatite-containing bone structures since it has never been found in elasmobranchs, whose skeleton is composed of calcified cartilage. Accordingly, recent work indicates that, in mammalian bone, BGP is required for adequate maturation of the hydroxyapatite crystal. Taken together, these data suggest that teleost fishes, presumably the first vertebrates to develop a BGP-containing skeleton, may be a useful model to further investigate BGP function. In addition, fish offer several advantages over mammalian models, due to a large progeny, external embryonic development and transparency of larvae. In the present work, the BGP cDNA and gene were cloned from a teleost fish, Sparus aurata, and its tissue distribution, pattern of developmental expression and evolutionary pathways analyzed. The molecular organization of the Sparus BGP (spBGP) gene is similar to mammalian BGP genes, and its expression throughout development follows the onset of calcification. The spBGP gene encodes a pre-propeptide of 97 amino acid residues, expressed only in bone and showing extensive homology to its mammalian homologs. Phylogenetic analysis of the available BGP sequences supports the hypothesis that all BGPs have a single origin and share a common ancestor with a related vitamin K dependent protein (Matrix Gla protein). PMID- 11404006 TI - Cloning and sequencing of human Eppin: a novel family of protease inhibitors expressed in the epididymis and testis. AB - In this report we describe the discovery of Eppin (Epididymal protease inhibitor), a gene on human chromosome 20 expressing three mRNAs encoding two isoforms of a cystine-rich protein containing both Kunitz-type and WAP-type four disuffide core protease inhibitor consensus sequences. Analysis of Eppin's genomic sequence from chromosome 20q12-13.2 predicts the existence of all three splice variants of Eppin and that all the exons conform to the AG/GT splicing rule. The presence of single bands on a Southern blot of human genomic DNA suggests that Eppin is a single copy gene. TATA box transcription initiation sites are present for both of the different Eppin 5' UTRs and examination of the promoter region 1800 bp upstream of the start codon revealed a number of putative transcription enhancer binding sites typical of genes expressed in the epididymis or testis. Northern blot and tissue specific PCR data indicate Eppin-1 is expressed only in the testis and epididymis; Eppin-2 is expressed only in the epididymis and Eppin-3 only in the testis. Antiserum prepared against recombinant EPPIN recognizes several strong bands on Western blots of human epididymal extracts from the caput and corpus regions. Immunohistochemistry indicates a strong pattern of expression by the ciliated cells of the efferent ducts and strong staining of ejaculated spermatozoa. Eppin represents the first member of a family of protease inhibitors characterized by dual inhibitor consensus sequences, both WAP-type and Kunitz-type consensus sequences. A second family member is predicted to exist on chromosome 20 approximately 4 kb downstream from Eppin's exon I, which has two WAP-type sequences and one Kumtz-type consensus sequence. PMID- 11404007 TI - Adaptable doxycycline-regulated gene expression systems for Drosophila. AB - We have engineered two new versions of the doxycycline (dox) inducible system for use in Drosophila. In the first system, we have used the ubiquitously expressed Drosophila actin5C promoter to express the Tet-Off transactivator (tTA) in all tissue. Induction of a luciferase target transgene begins 6 h after placing the flies on dox-free food. Feeding drug-free food to mothers results in universal target gene expression in their embryos. Larvae raised on regular food also show robust expression of a target reporter gene. In the second version, we have used the Gal4-UAS system to spatially limit expression of the transactivator. Dox withdrawal results in temporally- and spatially-restricted, inducible expression of luciferase in the adult head and embryo. Both the actin5C and Gal4-UAS versions produce more than 100-fold induction of luciferase in the adult, with virtually no leaky expression in the presence of drug. Reporter gene expression is also undetectable in larvae or embryos from mothers fed dox-containing food. Such tight control may be due to the incorporation of Drosophila insulator elements (SCS and SCS') into the transgenic vectors. These systems offer a practical, effective alternative to currently available expression systems in the Drosophila research community. PMID- 11404008 TI - Phylogeny and origin of 82 zygomycetes from all 54 genera of the Mucorales and Mortierellales based on combined analysis of actin and translation elongation factor EF-1alpha genes. AB - True fungi (Eumycota) are heterotrophic eukaryotic microorganisms encompassing ascomycetes, basidiomycetes, chytridiomycetes and zygomycetes. The natural systematics of the latter group, Zygomycota, are very poorly understood due to the lack of distinguishing morphological characters. We have determined sequences for the nuclear-encoded genes actin (act) from 82 zygomycetes representing all 54 currently recognized genera from the two zygomycetous orders Mucorales and Mortierellales. We also determined sequences for translation elongation factor EF 1alpha (tef) from 16 zygomycetes (total of 96,837 bp). Phylogenetic analysis in the context of available sequence data (total 2,062 nucleotide positions per species) revealed that current classification schemes for the mucoralean fungi are highly unnatural at the family and, to a large extent, at the genus level. The data clearly indicate a deep, ancient and distinct dichotomy of the orders Mucorales and Mortierellales, which are recognized only in some zygomycete systems. Yet at the same time the data show that two genera - Umbelopsis and Micromucor - previously placed within the Mortierellales on the basis of their weakly developed columella (a morphological structure of the sporangiophore well developed within all Mucorales) are in fact members of the Mucorales. Phylogenetic analyses of the encoded amino acid sequences in the context of homologues from eukaryotes and archaebacterial outgroups indicate that the Eumycota studied here are a natural group but provide little or no support for the monophyly of either zygomycetes, ascomycetes or basidiomycetes. The data clearly indicate that a complete revision of zygomycete natural systematics is necessary. PMID- 11404009 TI - Molecular cloning of a teleost growth hormone receptor and its functional interaction with human growth hormone. AB - This paper reports the first full-length cDNA sequence of a growth hormone receptor (GHR) from a teleost fish and its functional expression in cultured eukaryotic cells. The cDNA sequence, from Carassius auratus (goldfish), encodes a protein of 602 amino acids (aa) akin in architecture to the GHRs of other species. Despite the presence of motifs characteristic of GHR, the overall homology between the goldfish GHR and other GHRs is very low ( approximately 40% aa identity). CHO cells transfected with this receptor cDNA can be stimulated to proliferate by human growth hormone (hGH). In addition, the transfected cells can transactivate a co-expressed mammalian serine protease inhibitor (Spi) 2.1 promoter upon stimulation by hGH, indicating the successful interaction of the fish receptor with the mammalian ligand to evoke the down-stream post-receptor events. Tissue distribution studies indicated that the receptor is mostly expressed in the liver and hypothalamus of goldfish. A single mRNA transcript of a size of about 4 kb was found in the goldfish liver. PMID- 11404010 TI - A complex containing at least one zinc dependent HeLa nuclear protein binds to the intronic (gaa)(n) block of the frataxin gene. AB - We analyzed HeLa nuclear proteins binding to the (gaa)(n) harbouring intron 1 of nine frataxin alleles and characterized the structures of the repeats. Fragments with blocks longer than (gaa)(9) form spontaneously different intramolecular H-y topoisomeres in linear state. The observed triplexes depend on the length of the repeat. Interruption of the perfectly repeated (gaa)(n) block entails two structural regions. At least two HeLa nuclear proteins bind to the (gaa)(n) fragments resulting in a distinct major retarded complex as revealed by EMSA. One of these proteins is zinc dependent. Importantly, the fragment harbouring (gan)(121) binds additional proteins. Protein binding appears to be locus specific, and the binding affinity was found to be not random. The affinities of the different target fragments varied by a factor of four. Binding affinities of the fragments were not obviously correlated to differences in the composition of the repeats. DNase I footprinting revealed only weakly protected binding regions, but multiple HS sites in the repeat regions of the fragments. These findings and the fact, that DNA conformers observed in EMSA and electron microscopical experiments bind proteins, lead to the assumption that the proteins recognize, both, B-DNA and triple helical structures, but with different affinity. Possible functions of the proteins are discussed in the context of transformation of triple helical structures into B-DNA and the pathogenesis of FRDA. PMID- 11404011 TI - Gene structure, chromosomal localization, cDNA cloning and expression of the mouse ATP-gated ionotropic receptor P2X5 subunit. AB - The P2X receptors are oligomeric ligand-gated ion channels operated by extracellular ATP. Here we report the genomic and cDNA sequence of the mouse P2X(5) subunit, as well as its genomic organization, chromosomal localization and expression in select tissues. The mouse gene spans approximately 13 kb of DNA and contains thirteen exons. The cDNA encodes a 455 amino acid protein with 95% identity to the rat subunit. The P2X(5) subunit gene encodes a 2.6 kb mRNA that was found to in a number of tissues, with highest levels detected in heart and kidney. Results from rapid amplification of cDNA ends (RACE) PCR suggest that there are multiple transcriptional start sites located approximately 30-70 bp upstream from the start codon. The gene was localized to band B5 on Chromosome 11 using fluorescence in-situ hybridization (FISH), a region that has a high degree of synteny with human Chromosome 17. These results provide the initial information useful for further investigation into the functional role(s) of the P2X(5) subunit in physiological processes. PMID- 11404012 TI - p53 Pseudogene dating: identification of the origin of laboratory mice. AB - Mutations were accumulated with a wide variety in the p53 pseudogene of various wild mouse species and subspecies captured at different localities, as extensively observed in the exon 4 - exon 5 region. The rate of mutation accumulation in the mouse p53 pseudogene was estimated to be 1.4-2.1x10(-8) mutations/bp/year, which is 20-30 times faster than that of the functional p53 and makes the dating possible for the time range of 10(6) years or more. From comparison of the mutation spectrum, the origin of laboratory mice was identified to one of two M. m. domesticus groups. PMID- 11404013 TI - Molecular evolution of Sry and Sox gene. AB - The mammalian Sry on the short arm of the Y chromosome encodes a nuclear factor like protein harboring a DNA-binding domain known as the HMG box. The Sox genes encode similar factor like proteins, but the sequence similarity of the HMG box to that of Sry is variable as being at least 60%. The functional relationship of Sox to Sry genes with special reference to sex determination is unclear except for a few items such as human autosomal Sox9. Thus, it is significant to know more about the evolutionary in addition to the functional relationship between Sry and Sox genes for deepening and broadening our understanding concerning primary sex determination. Therefore, to clarify the ancestry and molecular evolution of the mammalian sex determining gene Sry with its evolutionary relationships to the Sox gene, a molecular phylogenetic tree for the HMG box superfamily was constructed and analyzed, and the following conclusions were reached: (1) The nuclear non histone HMG proteins are supposedly the oldest, appearing at least more than one billion years ago, before the divergence of animals and plants. They diverged into two subgroups: one contains HMG14 and HMG17, and the other one contains HMG1 and HMG2 with various other genes. Subsequent divergences include the nucleolar UBF, nuclear SSRP as well as fungal mating protein Mc, MAT and Ste11. (2) The Sox and Sry genes diverged following the diversification of lymphoid transcription factors TCF and LEF. The Sry gene might have definitely evolved from the Sox gene cluster a few hundred million years ago. Additionally, the marsupial Sry, e.g. from Wallabie's and Dunnart's, is distinguished by being distant from eutherian Sry, but being closely related to the Sox gene cluster. (3) Molecular evolutionary rates estimated in mammalian Sry as the divergent rate per 100 million years are much higher than in Sox genes or other genes from the HMG box superfamily. This rapid evolution of Sry might agree with the fact that the Srys are present not on the pseudoautosomal region but on the distal region with no recombination of the Y chromosomal short arm. PMID- 11404014 TI - Two distal Sp1-binding cis-elements regulate fibroblast growth factor receptor 1 (FGFR1) gene expression in myoblasts. AB - Skeletal myoblast cell proliferation and subsequent differentiation are dependent on developmentally regulated expression of the fibroblast growth factor receptor 1 (FGFR1) gene. We have previously reported the isolation and initial characterization of the chicken FGFR1 gene (cek1) promoter. Both distal and proximal regions of the promoter were identified as necessary for developmentally regulated transcriptional activity in proliferating myoblasts, including its down regulation in differentiated muscle fibers in vitro. Here we report detailed characterization of the molecular mechanism regulating FGFR1 promoter activity via the distal promoter region in proliferating myoblasts. This region was identified as a 242 base pair segment located greater than 1 kilobase upstream from the start of transcription that conferred increased transcriptional activity to a minimal thymidine kinase promoter. This segment contains two Sp1 binding sites. Site directed mutagenesis and transfection studies indicated that both Sp1 sites are functional and both are required for FGFR1 promoter activity. Furthermore, Sp1 binding to the two sites was synergistic enhancing FGFR1 promoter activity. The specificity of Sp1 binding to the two distal promoter cis elements was verified by electromobility shift and transfection assays employing an Sp1 expression construct. Differences in myoblast versus fibroblast-specific protein-DNA complex formation at these sites correlated with high promoter activity in myoblasts and significantly reduced promoter activity in fibroblasts. These studies for the first time establish a molecular mechanism regulating FGFR1 gene expression during myoblast proliferation. PMID- 11404015 TI - Introns resolve the conflict between base order-dependent stem-loop potential and the encoding of RNA or protein: further evidence from overlapping genes. AB - Many eukaryotic genes are split into exons and introns, the latter being removed post-transcriptionally so that only exon sequences appear in cytoplasmic RNAs. Since introns appear in both protein-encoding RNAs and non-protein-coding RNAs, they interrupt genetic information per se, not just protein-encoding information. A DNA sequence has the potential to carry more than one type of genetic information, but different types may conflict. Thus, it has been proposed that introns arose because sequences were unable to contain concomitantly complete information for the encoding both of stem-loops and of cytoplasmic products (protein and/or RNA). Stem-loop potential is held to be selectively advantageous since it promotes the recombination-dependent correction of genetic errors. Stem loop potential, the best local measure of which is base order-dependent stem-loop potential, tends to be less in exons than in introns. This is particularly evident in genes evolving rapidly under positive Darwinian selection, where the protein-encoding function is dominant. Evidence is now presented that the rare regions where genes overlap also impose excessive encoding demands so that the concomitant coding of base order-dependent stem-loop potential is decreased. Our results are consistent with the hypothesis that sequences with high stem-loop potential arose in the early 'RNA world'. Ancestors of modern genes would have entered this world when sequences (exons) encoding cytoplasmic products, were interspersed with sequences (introns) encoding selectively advantageous stem loops. Purine-loading pressure would also have favoured intron formation. PMID- 11404016 TI - Analysis of the enzymatic domains in the modular portion of the coronafacic acid polyketide synthase. AB - Coronafacic acid (CFA) is the polyketide component of coronatine (COR), a phytotoxin produced by the plant pathogen Pseudomonas syringae. The CFA polyketide synthase (PKS) consists of two open reading frames (ORFs) that encode type I multifunctional proteins and several ORFs that encode monofunctional proteins. Sequence comparisons of the modular portions of the CFA PKS with other prokaryotic, modular PKSs elucidated the boundaries of the domains that are involved in the individual stages of polyketide assembly. The two beta ketoacyl:acyl carrier protein synthase (KS) domains in the modular portion of the CFA PKS exhibit a high degree of similarity to each other (53%), but are even more similar to the KS domains of DEBS, RAPS, and RIF. Cfa6 possesses two acyltransferases- AT0, which is associated with a loading domain, and AT1, which uses ethylmalonyl-CoA (eMCoA) as a substrate for chain extension. Cfa7 contains an AT that uses malonyl-CoA as a substrate for chain extension. The Cfa6 AT0 shows 35 and 32% similarity to the DEBS1 and NidA1 AT0s, respectively, and 32 and 36% similarity to the Cfa6 and Cfa7 AT1s. Sequence motifs have previously been identified that correlate with AT substrates. The motifs in Cfa6 AT1 were found to correlate reasonably well with those predicted for methylmalonyl-CoA (mMCoA) ATs. The motifs in the AT of Cfa7 correlated more poorly with those predicted for MCoA ATs. Three ACP domains occur in the modular proteins of the COR PKS. The loading domain-associated ACP0 showed 38% similarity to the loading domain ACP0s of DEBS1 and NidA1 and 32-36% similarity to the two module-associated ACPs of the COR PKS. It exhibited a higher degree of similarity to the module-associated ACPs of RAPS. The two module-associated ACPs show 39% similarity to each other, but appear more closely related to module-associated ACP domains in RAPS and RIFS. Furthermore, the DH and KR domains of Cfa6 and Cfa7 show greater similarity to DH and KR domains in RAPS and RIFS than to each other. The CFA PKS includes a thioesterase domain (TE I) that resides at the C-terminus of Cfa7 and a second thioesterase, which exists as a separate ORF (Cfa9, a TE II). Analysis of a Cfa7 thioesterase mutant demonstrated that the TE domain is required for the production of CFA. The co-existence of TE domains within modular PKSs along with physically separated, monofunctional TEs (TE IIs) has been reported for a number of modular polyketide and non-ribosomal peptide synthases (NRPS). An analysis of the two types of thioesterases using Clustal X yielded a dendrogram showing that TE IIs from PKSs and NRPSs are more closely related to each other than to domain TEs from either PKSs or NRPSs. Furthermore, the dendrogram indicates that both types of TE IIs are more closely related to TE domains associated with PKSs than to TE domains in NRPSs. Finally, the overall % G+C content and the % G+C content at the third codon for all of the PKS genes in the COR cluster suggest that these genes may have been recruited from a gram-positive bacterium. PMID- 11404017 TI - Isolation and characterization of the murine transforming growth factor-beta2 promoter. AB - This report describes the isolation and characterization of the 5' flanking region of the murine transforming growth factor beta-2 (TGF-beta2) gene. A genomic clone containing the promoter region of the gene was isolated after screening a bacteriophage P1 genomic library. The resulting clone was sequenced and compared to promoters for the human and chicken TGF-beta2 genes. The sequence located near the transcription start site is highly conserved. It includes a TATA box, an E-box, and a largely conserved CRE/ATF site. A series of murine TGF-beta2 promoter/reporter constructs was generated to identify regulatory regions of the gene. As in the case of the human TGF-beta2 gene, sequences just upstream of the TATA box, including the CRE/ATF site, actively stimulate the murine TGF-beta2 promoter. However, unlike the human TGF-beta2 gene, the 5' flanking region of the murine TGF-beta2 gene contains a long alternating purine/pyrimidine repeat that unexpectedly exerts a strong positive effect on its promoter. This is of particular interest since alternating purine/pyrimidine repeats in other promoters have been observed to be inhibitory. PMID- 11404018 TI - Cloning, characterization and mapping of the mouse trehalase (Treh) gene. AB - Trehalase is the least studied of the membrane-bound alpha- glucosidase enzymes. Here we report the isolation and characterization of the mouse trehalase (Treh) gene. Initially, PCR using primers based on published rat cDNA sequence was used to clone a partial mouse cDNA. This allowed design of mouse primers which identified a single positive clone in a bacterial artificial chromosome (BAC) library of mouse genomic DNA. Analysis of BAC subclones showed that the Treh structural gene spans approximately 13 kb and comprises 15 exons. Data from genomic Southern blotting were consistent with mouse Treh being a single copy gene. The transcription initiation site was determined by both S1 nuclease mapping and 5' rapid amplification of cDNA ends (5' RACE) to be located 25 nt upstream of the ATG in exon 1. The mouse Treh exons were found to have an open reading frame of 1728 nt and the encoded protein of 576 amino acids showed 81, 82 and 93% amino acid sequence identity with rabbit, human and rat trehalase, respectively. The trehalase signature sequence found at amino acids 162 to 175 had 100% identity with the corresponding region of rabbit, human and rat and 79% identity with that for yeast trehalase. When a mouse Treh cDNA was used for Northern blot analysis of RNA from 12 mouse tissues, Treh mRNA expression was detected only in kidney and small intestine. The size of the mRNA in both of these tissues was estimated to be approximately 2.1 kb, furthermore both tissues appear to have the same transcription initiation site as determined by nuclease protection. Using the T31 radiation hybrid panel, mouse Treh was shown to be located on Chromosome 9 in a broad region that is orthologous with human Chromosome 11q23. The human trehalase gene (TREH) was identified in the latter location via database searching, which also revealed the overall structure of the human gene as being similar to that of the mouse. PMID- 11404019 TI - Genomic organization and characterization of the promoter region of murine GSTM2 gene. AB - In this study, we have isolated the genomic clone of the murine GSTM2 gene and determined its sequence. Consistent with the class mu genefamily, the mGSTM2 gene consists of eight exons. The exon-intron boundaries and the distribution of coding sequences within the exons of the known GST class mu family members were found to follow a similar pattern suggesting that various members of this family have originated from a single primordial gene by duplication and the structure has been closely maintained through evolution. By primer extension, the start of transcription was determined to be 40 bp upstream of the initial AUG codon. To gain an understanding of the mGSTM2 regulation, we have also cloned and analyzed its promoter region. Maximal activity was observed in a 170 bp 5'-flanking region. The activity was decreased by 3-fold in a 402 bp 5'-flanking region suggesting the presence of repressor elements. While no TATA box was identified, the presence of an SP1 site at position -38 was noted. Deletion of this SP1 site completely abrogated promoter activity. The promoter contained eight putative Myb responsive elements and its transcriptional activity was upregulated by t-Myb but not by c-Myb. PMID- 11404020 TI - Sequence of the E. coli O104 antigen gene cluster and identification of O104 specific genes. AB - The Escherichia coli O104 polysaccharide is an important antigen, which contains sialic acid and is often associated with EHEC clones. Sialic acid is a component of many animal tissues, and its presence in bacterial polysaccharides may contribute to bacterial pathogenicity. We sequenced the genes responsible for O104 antigen synthesis and have found genes which from their sequences are identified as an O antigen polymerase gene, an O antigen flippase gene, three CMP sialic acid synthesis genes, and three potential glycosyl transferase genes. The E. coli K9 group IB capsular antigen has the same structure as the O104 O antigen, and we find using gene by gene PCR that the K9 gene cluster is essentially the same as that for O104. It appears that the distinction between presence as group IB capsule or O antigen for this structure does not involve any difference in genes present in the O antigen gene cluster. By PCR testing against representative strains for the 166 E. coli O antigens and some randomly selected Gram-negative bacteria, we identified three O antigen genes which are highly specific to O104/K9. This work provides the basis for a sensitive test for rapid detection of O104 E. coli. This is important both for decisions on patient care as early treatment may reduce the risk of life-threatening complications and for a faster response in control of food borne outbreaks. PMID- 11404021 TI - Functional characterization of an ammonium transporter gene from Lotus japonicus. AB - NH(4)(+) is the main product of symbiotic nitrogen fixation and the external concentration of combined nitrogen plays a key regulatory role in all the different step of plant-rhizobia interaction. We report the cloning and characterization of the first member of the ammonium transporter family, LjAMT1;1 from a leguminous plant, Lotus japonicus. Sequence analysis reveals a close relationship to plant transporters of the AMT1 family. The wild type and two mutated versions of LjAMT1;1 were expressed and functionally characterized in yeast. LjAMT1;1 is transcribed in roots, leaves and nodules of L. japonicus plants grown under low nitrogen conditions, consistent with a role in uptake of NH(4)(+) by the plant cells. PMID- 11404022 TI - Characterization of a putative RND-type efflux system in Agrobacterium tumefaciens. AB - Sequencing of a 7277 bp fragment adjacent to the chvH locus of Agrobacterium tumefaciens revealed four open reading frames (ORFs), designated ameR, ameA, ameB and ameC, respectively. These ORFs exhibit amino acid similarities to components of Resistance-Nodulation-Cell Division (RND) type efflux systems. AmeA and AmeB show high homology to membrane fusion proteins (MFP) and RND-type transporters, whereas AmeC shows similarity to NodT and other members of outer membrane factor families. Mutations of the ameA and ameB genes did not affect the susceptibility profile of the wild-type strain to several detergents and antibiotics. In contrast, mutations of the ameC gene dramatically affected the susceptibility of the strain to these same inhibitory compounds. RT-PCR analysis demonstrated that the ameABC genes form an operon. In addition, ameC gene has its own promoter gene located in the intergenic region between ameB and ameC. Mapping upstream of ameA is ameR, which encodes a protein that shows similarity especially at its N terminal end to the TetR family of bacterial transcriptional regulators. AmeR negatively regulates expression of the ameABC operon. A mutation in ameR increased the resistance of the cells to several antimicrobial agents. This regulatory locus appears to be in the same operon as a gene located upstream which codes for a product that has high similarity to numerous 4-(N succinocarboxamide)-5-aminoimidazole ribonucleotide (SAICAR) synthetases. The possible role of the putative efflux pump coded by the ame genes is discussed. PMID- 11404023 TI - Cloning of genomic and cDNA for mouse isovaleryl-CoA dehydrogenase (IVD) and evolutionary comparison to other known IVDs. AB - Isovaleryl-CoA dehydrogenase (IVD) is an intramitochondrial homotetrameric flavoenzyme that catalyzes the conversion of isovaleryl-CoA to 3-methylcrotonyl CoA in the leucine catabolism pathway. Deficiency of IVD in humans causes isovaleric acidemia, which shows tremendous clinical variability for reasons that are unknown. To help better understand this disorder, we have cloned and sequenced the mouse IVD genomic and cDNAs. The mouse IVD gene spans approximately 17 kb and contains 12 coding exons organized identically to the human gene. It maps to mouse chromosome 2 in the area of band 2E4-E5, corresponding to the syntenic region of human chromosome 15. Mouse IVD predicted amino acid sequences are 95.8 and 89.6% identical to that of the rat and human sequences, respectively, with conservation of key functional residues. We have now identified IVD sequences from seven species. Comparison of these sequences shows that the rat and mouse proteins are the most closely related, both of which, in turn, share highest homology to human. All of the mammalian enzymes appear to be more closely related than any of the IVDs on other branches of the phylogram, while the fly and worm IVDs are the most divergent. The invertebrate IVDs are more closely related to the mammalian enzymes than to those from two plant species. PMID- 11404024 TI - A novel gene that is up-regulated during recovery from cold shock in Drosophila melanogaster. AB - Gene expression during recovery at 25 degrees C (rearing temperature) after cold shock (0 degrees C) was studied in Drosophila melanogaster using a subtraction technique. A novel gene (Frost, abbreviated as Fst) was considerably up-regulated during recovery after cold shock. In addition, a prolongation of cold shock was more effective for induction. In contrast to cold shock, Fst gene did not respond to heat shock. This gene is apparently the same as the unidentified gene, CG9434. Fst has high internal repeats not only in nucleotide but also in amino acid sequences. In addition, FST protein has a proline-rich region. The deduced amino acid sequence revealed a modular structure; i.e., a signal peptide in the N terminal region followed by a long hydrophilic region. Therefore, this protein is likely to be directed into ER and secreted into extracellular space. PMID- 11404025 TI - Molecular mechanisms of human single-minded 2 (SIM2) gene expression: identification of a promoter site in the SIM2 genomic sequence. AB - We previously postulated that the single-minded 2 (SIM2) gene identified on the human chromosome 21q22.2 is a good candidate gene for the pathogenesis of mental retardation in Down syndrome because its mouse homolog exhibits preferential expression in the mouse diencephalon during early embryogenesis. We analyzed the genomic sequence of the entire SIM2 gene which consists of 11 exons and spans over 50 kb. As a step toward understanding the molecular mechanisms of SIM2 gene expression, we have analyzed the human SIM2 gene expression in nine established human cell lines. Three transcripts of 3.6, 4.4, and 6.0 kb were detected in the glioblastoma cell line, T98G, neuroblastoma cell line, TGW, and transformed embryonic kidney cell line, 293. The RACE analysis using SIM2-expressing human cell line T98G provided evidence for the transcription start site at approximately 1.2 kb upstream of the translation initiation site. The transfection assay using various deletion constructs with reporter gene suggested the presence of a presumptive promoter region. Transient transfection assay in T98G cell line revealed a significant promoter activity located in the 60 bp sequence between nt -1385 and -1325 upstream region of the translation initiation site. This 60 bp sequence contains cis-elements for c-myb, E47 and E2F transcription factors. Moreover, the gel retardation assay using oligo-DNA of various cis-element sequences indicated the presence of protein factor(s) which bind to the cis-element for c-myb. These results suggested that binding of a protein transcription factor(s) such as c-myb or that alike regulates transcription of the SIM2 gene by binding to a small upstream region. PMID- 11404026 TI - Organization, expression, and localization of the murine mdmx gene and pseudogene. AB - The mdmx gene is the first additional member of the mdm2 gene family to be isolated. It encodes a protein similar to MDM2 in several domains and also retains the ability to bind and inhibit p53 transactivation in vitro. However, mdmx does not appear to be transcriptionally regulated by p53. We have cloned and characterized the murine mdmx genomic locus from a 129 genomic library. The mdmx gene contains 11 exons, spans approximately 37 kb of DNA, and is located on mouse chromosome 1. The genomic organization of the mdmx gene is identical to that of mdm2 except at the 5' end of the gene near the p53 responsive element. Additionally, a pseudogene for mdmx was also identified that resides on the mouse X chromosome. Expression analysis of mdmx transcripts during mouse embryogenesis revealed constitutive and ubiquitous expression throughout development. PMID- 11404027 TI - In vivo evaluation of hydrochlorothiazide liquisolid tablets in beagle dogs. AB - This study was carried out to evaluate the absorption characteristics of experimentally developed hydrochlorothiazide liquisolid tablets using six male beagle dogs. Comparison with reference commercial tablets was made. As no bibliographic data were found for the pharmacokinetic parameters of the drug in dogs, an intravenous drug administration was included in the study. The drug was administered orally as a single 25 mg dose of commercial and liquisolid tablets on two occasions in a randomized two-way crossover design. The pharmacokinetic parameters of the drug post intravenous dosing were reported for the first time. The results of the oral administration revealed statistically significant differences between the liquisolid and the commercial tablets in the area under the plasma concentration-time curve, the peak plasma concentration, and the absolute bioavailability. On the other hand, no significant differences were observed between the two formulations with regard to the mean residence time, the mean absorption time, and the rate of absorption. The absolute bioavailability of the drug from the liquisolid tablets was 15% higher than that from the commercial one. The parametric 90% confidence intervals for the different parameters were higher than the commonly expected intervals for bioequivalency, indicating greater bioavailability of the liquisolid tablets. PMID- 11404028 TI - Influence of hydrogel structure on the processes of water penetration and drug release from mixed hydroxypropylmethyl cellulose/thermally pregelatinized waxy maize starch hydrophilic matrices. AB - The kinetics of water penetration and molsidomine release from both hydroxypropylmethyl cellulose (HPMC) and mixed HPMC/thermally pregelatinized waxy maize starch (SDWMT) hydrophilic matrices has been examined in 0.1 mol x dm(-3) HCl (pH 1.0) and 0.06 mol x dm(-3) Na3PO4/HCl buffer (pH 6.8). The rheological oscillatory test parameters of their gel layers obtained by swelling of the matrices in the two aqueous media have been observed. The kinetic swelling properties of mixed HPMC/SDWMT hydrogels (i.e. degree and velocity of both water penetration and swelling, transport mechanism which controls solvent sorption) directly influence the drug release behaviour and the structural features of the formed gel layer. Both diffusion processes are diffusion-controlled ones, their mechanisms being influenced insignificantly by the relaxation properties of the hydrated macromolecules. It has been established by means of comparative viscoelastic analysis, that mixed HPMC/SDWMT hydrogels demonstrate the typical behaviour of 'filled' composite systems having poor adhesion between the surface of the elastic SDWMT 'filler' and the continuous HPMC phase. Due to the inter phase relations between the swollen starch granules and the linear cellulose derivative as well as to the specific structure of amylopectin molecule, the pregelatinized waxy maize starch shows a stronger influence on the velocities of both water penetration and drug release from mixed HPMC/SDWMT matrices. PMID- 11404029 TI - Paclitaxel loaded poly(L-lactic acid) microspheres: properties of microspheres made with low molecular weight polymers. AB - Microspheres were prepared from poly(L-lactic acid) polymers having molecular weights between 500 and 50k g/mol. The polymers were synthesized using two initiator molecules, L-lactic acid oligomer (PLLA-LA) or stearyl alcohol (PLLA SA). For both PLLA-LA and PLLA-SA polymers, glass (Tg) and melting (Tm) transition temperatures and enthalpy of melting all increased as the polymer molecular weight increased. PLLA-SA showed the greatest change in Tg (-13 to 54 degrees C) as molecular weight increased from 500 to 10k x g/mol, compared to 25 to 55 degrees C for PLLA-LA polymers. Changes in Tm and enthalpy of melting with increasing molecular weight were similar for both PLLA-LA and PLLA-SA. Paclitaxel release from 30% paclitaxel loaded microspheres in the size range of 50-90 microm was affected by these changes in polymer properties as molecular weight increased. As the molecular weight increased from 2k to 50k x g/mol the amount of drug released from microspheres over 14 days decreased from 76 to 11% of the initial drug load. The release profiles were consistent with a diffusion controlled mechanism provided a two-compartment model was employed. According to this model, the total amount of 'available' drug (compartment 1) was released by diffusion in 14 days while the remainder (compartment 2) was confined within the polymeric matrix and could not diffuse out at a measurable rate. Following the in vitro release study, microsphere made from 2k-10k g/mol polymers showed significant signs of disintegration whereas 50k x g/mol polymer microspheres remained intact. PMID- 11404030 TI - Preparation and characterization of two-phase melt systems of lidocaine. AB - The melting point of lidocaine was significantly lowered when mixed with thymol and/or aqueous ethanol. Mixtures of lidocaine and thymol at ratios within the range of 30:70-70:30 (w:w) became homogeneous oils at 25 degrees C. In a pH 9.2 carbonate buffer containing 25% ethanol, lidocaine (5%, w:w) also liquefied at 25 degrees C. The studies led to the development of novel two-phase melt systems of lidocaine (TMS) which consisted of a highly concentrated oil phase of lidocaine and an alcoholic aqueous phase. A compositional phase diagram showed that in aqueous dispersions of lidocaine, concurrent use of thymol and ethanol depressed the melting point of lidocaine more effectively than when they were used individually. Both thymol and aqueous ethanol were necessary as melting point depressing agents to achieve the highest possible lidocaine concentration of 87% (w:w) in the oil phase of a TMS at 25 degrees C. Containing an internal oil phase and an external aqueous phase at ambient temperature, such a TMS can be readily formulated into topical O/W cream after addition of proper surfactants and thickening agents. In an anesthetic activity test using mouse tail-flick model, a 5% lidocaine cream prepared was highly effective as shown by the prolonged latency time of the mice to a heat stimulus as compared with a placebo (P<0.05). PMID- 11404031 TI - Inclusion complex of piroxicam with beta-cyclodextrin and incorporation in cationic microemulsion. In vitro drug release and in vivo topical anti inflammatory effect. AB - Topical formulations of piroxicam were evaluated by determination of their in vitro release and in vivo anti-inflammatory effect. The in vitro release assay demonstrated that the microemulsion (ME) systems provided a reservoir effect for piroxicam release. However, the incorporation of the ME into carboxyvinilic gel provoked a greater reduction in the release of piroxicam than the ME system alone. Anti-inflammatory activity was carried out by the cotton pellet granuloma inhibition bioassay. Topical anti-inflammatory effect of the piroxicam inclusion complex/ME contained in carboxyvinilic gel showed significant inhibition of the inflammation process (36.9%, P<0.05). Subcutaneous administration of the drug formulations showed a significant effect on the inhibition of inflammation, 68.8 and 70.5%, P<0.05, when the piroxicam was incorporated in ME and in the combined system beta-cyclodextrin (beta-CD)/ME, respectively, relative to the buffered piroxicam (42.2%). These results demonstrated that the ME induced prolonged effects, providing inhibition of the inflammation for 9 days after a single dose administration. PMID- 11404032 TI - Effect of phosphatidylcholine on skin permeation of indomethacin from gel prepared with liquid paraffin and hydrogenated phospholipid. AB - The effects of hydrogenated and unhydrogenated phosphatidylcholine (HPC, PC) on the permeation of indomethacin (IM) through hairless rat skin were investigated using liquid paraffin (LP) and a gel prepared with LP and hydrogenated soybean phospholipid (HSL). IM solubility at 95 degrees C increased in proportion to the concentration of HPC or PC, whereas solubility at 37 degrees C did not increase with HPC. IM showed no permeation until 10 h from LP without HPC/PC, but permeated at rates of approximately 5 and 10 microg/cm2 within 10 h from LP with HPC and PC, respectively. The permeation from the gel with various formulations (HSL, 15%; PC/HPC, 0-5%; IM, 0.5-2%) was determined. Permeation rates were 1.7 4.8 microg/cm2 per h and were proportional to the skin concentration. Skin concentration was correlated to the release rate from the gel. We concluded that IM was solubilized by phospholipids, high activity in the vehicle led to high partition of IM in skin, and permeation increased due to a high skin concentration. PMID- 11404033 TI - Granule deformation and densification during compression of binary mixtures of granules. AB - The purpose of this study was to investigate whether the deformation and densification during compression of one type of granules are affected by adjacent granules of a different porosity, corresponding to different mechanical strength. Three mixtures were prepared, each consisting of two types of microcrystalline cellulose pellets (intermediate porosity study pellets plus low, intermediate or high porosity surrounding pellets) in the proportion 1:7. The mixtures were compressed and the study pellets were retrieved and analysed in terms of porosity, thickness, surface area and shape. It was shown that the study pellets were compressed by deformation and densification. The degree of densification (decrease in porosity) of the study pellets was independent of the porosity of the surrounding pellets but the deformability (changes in the thickness, surface area and shape) of the study pellets was linked with the porosity of the surrounding pellets. It is concluded that the mode of deformation of the study pellets was regulated by the porosity of the surrounding granules; in a mixture containing granules with a low porosity, compression resulted in irregular study granules with regularly positioned indentations caused by the surrounding granules. The compression properties of the surrounding granules affected the flattening of the study granules to a lesser degree. PMID- 11404034 TI - Permeability of lipophilic compounds in drug discovery using in-vitro human absorption model, Caco-2. AB - Highly lipophilic compounds are often encountered in the early stages of drug discovery. The apparent permeability (Papp) of these compounds in Caco-2 cell could be underestimated because of considerable retention by the Caco-2 monolayer and non-specific binding to transwell surface. We have utilized a general approach for the determination of permeability of these compounds, which includes the addition of 1-5% DMSO in the apical (AP) and 4% bovine serum albumin (BSA) in the basolateral (BA) side. Two highly lipophilic and highly protein bound Schering compounds, SCH-A and SCH-B, exhibited poor recovery and low Papp in the conventional Caco-2 system that included 1% DMSO in the AP and BA sides. In contrast, both compounds were well absorbed in cynomolgus monkeys. Inclusion of BSA (up to 4%) in the BA side provided necessary absorptive driving force similar to in vivo sink conditions improving both recovery and Papp of these compounds as well as progesterone, a model highly lipophilic and highly protein bound compound. Whereas, the recovery and Papp of mannitol (high recovery, low permeability) and propranolol (high recovery, high permeability) remained unaffected. The presence of 4% BSA increased Papp of SCH-A, SCH-B, and progesterone by five-, four-, and three-fold, respectively. We also compared this approach with a second, based on the disappearance of the compound from the AP side, which resulted in a reasonable estimate of the permeability (23.3x10(-6) cm/s) for SCH-A. The results demonstrated that the reliable estimates of permeability of highly lipophilic compounds that are subjected to considerable retention by the cell monolayer and exhibit non-specific binding are obtained by the addition of BSA to the BA side. PMID- 11404035 TI - Stabilization of minodronic acid in aqueous solution for parenteral formulation. AB - The composition, concentration, and buffer pH of potential minodronic acid formulations were evaluated for their drug stability and for their tendency to generate particles after storage for up to 4 weeks at 60 degrees C. The results indicate that citrate and tartrate buffers maintain drug stability and inhibit the formation of particles. The stability of minodronic acid in these solutions increased slightly as the buffer concentration increased, exhibiting less particle formation than in other buffers. Since citrate buffer was considered the most promising stabilizer for minodronic acid, the pH-stability relationship in 100 mM citrate with pH ranging from 3 to 7 was evaluated during storage for 4 weeks at 60 degrees C. The results demonstrate that solution pH of 3-5 result in optimal stability of minodronic acid with no formation of precipitates. A white precipitate was observed in citrate-containing sample solutions with pH of 6 and 7. Analysis of the isolated precipitate provided support for the hypothesis that the precipitate is a complex between minodronic acid and aluminum ions apparently leached from the glass of the ampoules. PMID- 11404036 TI - Delivery of HFA and CFC salbutamol from spacer devices used in infancy. AB - The aim of the study was to compare the in vitro delivery of four salbutamol pressurized metered-dose inhalers (pMDIs) via the three spacer devices commonly used in European infants: Aerochamber-Infant, Babyhaler, and metallic NES-spacer. Emitted dose (ED) and fine particle dose (FPD, particles<5.8 microm) of each combination of spacer device and pMDI (chlorofluorocarbon-based Ventoline, Eolene, Spreor, and hydrofluoroalkane-based Airomir were measured respectively using unit dose sampling tubes (n=30 per combination) and an 8-stage cascade impactor (n=6 per group). The results were compared by analysis of variance and the Student-Newman-Keuls method. ED of Airomir was always greater than for Ventoline (P<0.05). FPD obtained with Ventoline was the lowest, with Eolene>Airomir=Spreor>Ventoline (P<0.05). Only Airomir produced a similar FPD with all three spacer devices. Chlorofluorocarbon-salbutamol pMDIs are not generics when used with spacer devices. The three spacer devices may be used interchangeably with Airomir. PMID- 11404037 TI - Bioadhesive starch microspheres and absorption enhancing agents act synergistically to enhance the nasal absorption of polypeptides. AB - This paper investigates the effect of starch microspheres on the absorption enhancing efficiency of various enhancer systems in formulations with insulin after application in the nasal cavity of sheep. The enhancers studied were lysophosphatidylcholine, glycodeoxycholate and sodium taurodihydroxyfusidate, a bile salt derivative. The enhancers were selected on the basis of their perceived or proven mechanism of action and worked predominantly by interacting with the lipid membrane. The bioadhesive starch microspheres were shown to increase synergistically the effect of the absorption enhancers on the transport of the insulin across the nasal membrane. Dependent on the potency of the enhancer system the increment in absorption enhancement was shown to be from 1.4 times to 5 times that obtained for the absorption enhancer in solution. PMID- 11404038 TI - A TSM sensor investigation of low crystallinity cellulose films. AB - A thickness shear mode (TSM) quartz sensor has been used to characterize the substantivity, viscoelasticity, and mucoadhesive properties of low crystallinity cellulose (LCC) films. LCC is a novel pharmaceutical excipient that has been attributed with mucoadhesive properties. Thin films of LCC were deposited onto TSM sensors by a spin coating technique. The films were treated by passing water or 1.0% w/v mucin solution (pH 3.7 or 7.0) over the surface. Changes in the mass and viscosity of the film were observed by monitoring changes in the impedance spectra of the coated TSM sensors. Scanning electron micrographs (SEMs) of each film were used to assist the interpretation of the TSM sensor data. This study showed that LCC forms highly tenacious and viscoelastic films able to withstand prolonged (approximately 1 h) exposure to both water and mucin solution. Furthermore, these results indicate that the films may have mucoadhesive properties as LCC was found to bind significant (P<0.05) amounts of mucin in comparison with control measurements. Mucin binding to the LCC sensor was greater at pH 3.7 (P<0.05) than at pH 7.0, suggesting that the LCC formulation is mucoadhesive under these conditions. PMID- 11404039 TI - Processing of carbamazepine-PEG 4000 solid dispersions with supercritical carbon dioxide: preparation, characterisation, and in vitro dissolution. AB - The purpose of this study was to apply the attractive technique of the supercritical fluid to the preparation of solvent-free solid dispersions. In particular, the gas antisolvent crystallisation technique (GAS), using supercritical carbon dioxide as processing medium, has been considered to prepare an enhanced release dosage form for of the poorly soluble carbamazepine, employing PEG 4000 as a hydrophilic carrier. The physical characterisation of the systems using laser granulometer, powder X-ray diffraction, thermal analyses, and scanning electron microscopy was carried out in order to understand the influence of this technological process on the physical status of the drug. The results of the physical characterisation attested a substantial correspondence of the solid state of the drug before and after treatment with GAS technique, whereas a pronounced change in size and morphology of the drug crystals was noticed. The dramatic reduction of the dimensions and the better crystal shape, together with the presence of the hydrophilic polymer determined a remarkable enhancement of the in vitro drug dissolution rate. PMID- 11404040 TI - A comparison of alternative polymer excipients and processing methods for making solid dispersions of a poorly water soluble drug. AB - Solid dispersions were prepared with the extremely poorly water soluble drug, probucol and the water soluble polymers, polyvinyl pyrrolidone (PVP), polyacrylic acid (PAA) or polyethylene oxide (PEO) and blends of these polymers. The solid dispersions were prepared either by the solvent evaporation method, or by compression moulding into films. The materials were characterised by a combination of thermal analysis and FT-Raman spectroscopy. The physical state of the drug was observed to be dependent on the carrier, thus the PVP solid dispersions contained amorphous probucol, whilst the PAA and PEO systems contained the crystalline polymorph II. The method of production was not found to greatly influence the state of the drug in the solid dispersion. The greatest extent of release into solution was observed for the binary blend of drug and PEO, and the blending of polymers was not found to have any advantageous effects in this study. PMID- 11404041 TI - Stripping method to quantify absorption of two sunscreens in human. AB - With the aim to know the remanence of two sunscreens, PEG-25 PABA and benzophenone, two cases have been considered: the application of aqueous solution of two filters studied for the application of oil-in-water (O/W) emulsions containing the same two filters on the skin of 21 voluntary women. In order to determine the quantity of filter remaining in the stratum corneum after times of application, which vary from 1 to 7 h, a series of six strippings have been carried out. The sunscreen agents were assessed by HPLC. With this study, it has been possible to emphasize the best remanence of benzophenone 4. Moreover, the incorporation of photo-protective agents in a O/W emulsion promotes their penetration, and this is particularly due to benzophenone 4. PMID- 11404042 TI - Experimental research on ageing in Russia. AB - An analysis of the current situation in Russian biogerontology is presented in this paper. There are several active groups in Russia pursuing research in biogerontology capable of producing results publishable in international journals of high repute. The main directions of research on the biology of ageing in this country are prevention of premature ageing, the role of free radicals and of the endocrine system (in particular, the pineal gland) in the mechanisms of ageing, carcinogenesis and ageing and population genetics of ageing. Several groups are conducting fruitful research on the theoretical aspects of the biology of ageing. Only a few teams are focusing on molecular biology and the genetics of ageing. In the past few years, many more researchers in fields highly relevant to gerontology have been attracted by issues in gerontological research. In Russia, the most basic problem facing researchers in biogerontology and other relevant areas is an almost complete absence of support from the State and other decision makers. PMID- 11404043 TI - Basic gerontology research in Ukraine: main trends. AB - The main trends of basic research in gerontology in Ukraine are presented in this paper. The greatest attention is paid to the directions of research which have been traditionally followed in Ukrainian gerontology, such as neurohumoral regulation of the activities of different organs and systems and mechanisms of its age-related changes, metabolism in ageing, age-related immunology, elaboration of the ways and approaches to life-span prolongation. In recent years, more attention has been paid to research into the molecular--genetic mechanisms of ageing, the development of genetic and gene-regulatory therapy methods, the study of the mechanisms of accelerated and, above all, radiation induced ageing and the elaboration of prophylactic methods against it. PMID- 11404044 TI - Oxidative damage to mitochondria and aging. AB - Oxidative damage has been implicated to be a major factor in the decline in physiologic function that occurs during the aging process. Because mitochondria are a primary site of generation of reactive oxygen species, they have become a major focus of research in this area. Increased oxidative damage to mitochondrial proteins, lipid and DNA has been reported to occur with age in several tissues in a variety of organisms. Decreased activity of electron transport chain complexes and increased release of reactive oxygen species from the mitochondria with age suggest that alterations in mitochondrial function occur with age as a consequence of increased oxidative damage. In addition, age-related alterations in the mitochondrial pathway of apoptosis, which could have profound affects on the physiological function of a tissue, could arise from oxidative damage to mitochondria. Alterations in mitochondrial turnover with age could also contribute to an increase in the number of dysfunctional mitochondria with age. PMID- 11404045 TI - Smooth muscle cell apoptosis in arteriosclerosis. AB - Arteriosclerosis, a paradigmatic age-related disease, encompasses (spontaneous) atherosclerosis, restenosis after percutaneous transluminal coronary angioplasty, autologous arterial or vein graft arteriosclerosis and transplant arteriosclerosis. In all types of arteriosclerosis, vascular smooth muscle cell (SMC) accumulation in the intima is a key event, but abundant evidence also indicates the importance of SMC apoptosis in the development of arteriosclerosis. Because SMC proliferation and apoptosis coincide in arteriosclerotic lesions, the balance between these two processes could be a determinant during vessel remodeling and disease development. Various stimuli, including oxidized lipoproteins, altered hemodynamic stress and free radicals, can induce SMC apoptosis in vitro. As risk factors for arteriosclerosis, these stimuli may also lead to vascular cell apoptosis in vivo. The presence of apoptotic cells in atherosclerotic and restenotic lesions could have potential clinical implications for atherogenesis and contributes to the instability of the lesion. Based on the progress in this field, this review focuses on the mechanism and impact of SMC apoptosis in the pathogenesis of arteriosclerosis and highlights the role of biomechanical stress in SMC apoptosis. PMID- 11404046 TI - The second European Congress on Biogerontology: from molecules to humans. PMID- 11404047 TI - The programmed death phenomena, aging, and the Samurai law of biology. AB - Analysis of the programmed death phenomena from mitochondria (mitoptosis) to whole organisms (phenoptosis) clearly shows that suicide programs are inherent at various levels of organization of living systems. Such programs perform very important functions, purifying (i) cells from damaged (or unwanted for other reasons) organelles, (ii) tissues from unwanted cells, (iii) organisms from organs transiently appearing during ontogenesis, and (iv) communities of organisms from unwanted individuals. Defence against reactive oxygen species (ROS) is probably one of primary evolutionary functions of programmed death mechanisms. So far, it seems that ROS play a key role in the mito-, apo-, organo- and phenoptoses. Here a concept is described which tries to unite Weismann's concept of aging as an adaptive programmed death mechanism and the alternative point of view considering aging as an inevitable result of accumulation in an organism of occasional injuries. It is suggested that injury accumulation is monitored by special system sending a death signal to actuate a phenoptotic program when the number of injuries reaches some critical level. The system in question is organized in such a way that the lethal case appears to be a result of phenoptosis long before occasional injuries make the functioning of the organism impossible. This strategy is supposed to prevent the appearance of asocial monsters capable to ruining kin, community and entire population. These relationships are regarded as an example of the Samurai law of biology: 'It is better to die than to be wrong'. It is stressed that for humans these cruel regulations look like an atavism that should be overcome to prolong the human life span. PMID- 11404048 TI - Strategy for identifying biomarkers of aging in long-lived species. AB - If effective anti-aging interventions are to be identified for human application, then the development of reliable and valid biomarkers of aging are essential for this progress. Despite the apparent demand for such gerotechnology, biomarker research has become a controversial pursuit. Much of the controversy has emerged from a lack of consensus on terminology and standards for evaluating the reliability and validity of candidate biomarkers. The initiation of longitudinal studies of aging in long-lived non-human primates has provided an opportunity for establishing the reliability and validity of biomarkers of aging potentially suitable for human studies. From the primate study initiated in 1987 at the National Institute on Aging (NIA), the following criteria for defining a biomarker of aging have been offered: (1) significant cross-sectional correlation with age; (2) significant longitudinal change in the same direction as the cross sectional correlation; (3) significant stability of individual differences over time. These criteria relate to both reliability and validity. However, the process of validating a candidate biomarker requires a greater standard of proof. Ideally, the rate of change in a biomarker of aging should be predictive of lifespan. In short-lived species, such as rodents, populations differing in lifespan can be identified, such as different strains of rodents or groups on different diets, such as those subjected to calorie restriction (CR), which live markedly longer. However, in the NIA primate study, the objective is to demonstrate that CR retards the rate of aging and increases lifespan. In the absence of lifespan data associated with CR in primates, validation of biomarkers of aging must rely on other strategies of proof. With this challenge, we have offered the following strategy: If a candidate biomarker is a valid measure of the rate of aging, then the rate of age-related change in the biomarker should be proportional to differences in lifespan among related species. Thus, for example, the rate of change in a candidate biomarker of aging in chimpanzees should be twice that of humans (60 vs 120 years maximum lifespan); in rhesus monkeys three times that of humans (40 vs 120 years maximum lifespan). The realization of this strategy will be aided by developing a primate aging database, a project that was recently launched in cooperation with the NIA, the National Center for Research Resources, and the University of Wisconsin Regional Primate Research Center. PMID- 11404049 TI - Critical assessment of paradigms in aging research. PMID- 11404050 TI - Stress, DNA damage and ageing -- an integrative approach. AB - Ageing is highly complex, involving multiple mechanisms at different levels. Nevertheless, recent evidence suggests that several of the most important mechanisms are linked via endogenous stress-induced DNA damage caused by reactive oxygen species (ROS). Understanding how such damage contributes to age-related changes requires that we explain how these different mechanisms relate to each other and potentially interact. In this article, we review the contributions of stress-induced damage to cellular DNA through (i) the role of damage to nuclear DNA and its repair mediated via the actions of poly(ADP-ribose) polymerase-1, (ii) the role of damage to telomeric DNA and its contribution to telomere-driven cell senescence, and (iii) the role of damage to and the accumulation of mutations in mitochondrial DNA. We describe how an integrative approach to studying these mechanisms, coupled with computational modelling, may be of considerable importance in resolving some of the complexity of cellular ageing. PMID- 11404051 TI - Increase of homozygosity in centenarians revealed by a new inter-Alu PCR technique. AB - In the present study a novel inter-Alu PCR technique that allows one to detect inter-individual differences in the genomic regions flanked by Alu repetitive sequences was developed. Two primers complementary to sequences present in different Alu repeats and marked with two different fluorochromes were used in the same PCR reaction, and the PCR products were separated and analyzed by capillary electrophoresis using an automatic sequencer. The method is highly reliable, and three patterns of peaks (QM376-400, QM780-790 and QM480) appeared to be representative for germ-line polymorphisms, as suggested by the results obtained in nine couples of monozygotic twins and four three-generation families. The frequency of these polymorphic peaks was studied in two different age groups (100 young subjects and 69 centenarians). In two out of the three regions (QM376 400 and QM480) a significant increase in homozygote genotypes frequency was observed in centenarians. These counterintuitive results suggest that increased homozygosity contributes to human longevity. This novel inter-Alu PCR approach could represent a valuable tool to identify longevity-associated DNA sequences interspersed throughout human genome, without making any a priori assumption about their nature and function. PMID- 11404052 TI - Hormonal changes in aging men: a therapeutic indication? AB - The rise in male life expectancy is paralleled by increased age-related clinical signs and symptoms such as muscle weakness, osteoporosis, benign prostatic hyperplasia, changes in body composition, fatigue, decreased sexual interest and activity, and increased prevalence of erectile dysfunction, all of which limit the quality of life. Many of these symptoms are similar to those of clinically well-defined hormone deficiencies, e.g. Kallman syndrome, Prader--Labhart--Willi syndrome or deficiencies due to treatment of pituitary tumors. Three male endocrine axes are characterized by age-related changes in concentrations of circulating hormones: (i) the hypothalamic--pituitary--testicular axis with lower serum levels of testosterone (T) and higher serum levels of luteinizing and follicle-stimulating hormone, (ii) the hypothalamic--pituitary--adrenal axis with its gradual decline in dehydroepiandrosterone (DHEA) and DHEA-sulfate (DHEAS), (iii) the growth hormone (GH)/insulin-like growth factor I (IGF-I) axis showing decreased hormone production concomitant with symptoms similar to those of GH deficient adults. The beneficial effects of hormone replacement in nonelderly hormone-deficient individuals and in postmenopausal women raised hope that hormone substitution might prevent or even reverse some of the symptoms of male aging. However, this approach is hampered by the lack of individual age-related hormone reference values and reliable clinical read-out parameters. The findings so far do not support the need for widespread hormone replacement in elderly men. Larger long-term prospective studies are needed to identify clinically useful read-out parameters, and then demonstrate that hormone replacement can translate into functional parameters, thereby providing the individual benefit of treatment for aging men. PMID- 11404053 TI - Human aging and melatonin. Clinical relevance. AB - Melatonin is a hormone produced mainly by the pineal gland and secreted primarily at night, when it reaches levels 10 times higher than those present in the daytime. The highest melatonin levels are found in children younger than 4 yr; thereafter melatonin levels begin to decline with age. As a chronobiotic, melatonin acts on sleep by phase-advancing or delaying the sleep--wake cycle so that sleep onset occurs earlier or later than usual. Beneficial effects of melatonin have been observed in delayed and advanced sleep phase syndromes. These effects depend on the time that the hormone is administered. Melatonin is also used for jet lag and has been tried in shift workers and night workers to re entrain their desynchronized rhythms. Melatonin also has free radical-scavenging properties that have primarily been observed in vitro at pharmacological concentrations. PMID- 11404054 TI - Life span extension and cancer risk: myths and reality. AB - A significant increase in the number of old people in the populations of developed countries was followed by an increase in morbidity and mortality resulting from main age-related diseases -- cardiovascular, cancer, neurodegenerative, diabetes mellitus, decrease in resistance to infections. Obviously, the development of the means of prevention of the premature aging of humans is crucial for the realization of this program. However, data available on such kind of means are rather scarce, contradictory and are often not reliable from the points of view of the adequacy of the experiments to current scientific requirements as well as the interpretation of the results and safety. Data available on the increase in life span and the adverse effects of the following geroprotectors were critically analyzed: antioxidants, chelate agents and lathyrogens, succinate, adaptogens and herbs, neurotropic drugs, inhibitors of monoamine oxidase, glucocorticoids, dehydroepiandrosterone, sex and growth hormones, melatonin, pineal peptide preparations, protein inhibitors, antidiabetic biguanides, thymic hormones and peptides, immunomodulators, enteroadsorbents, lypofuscin inhibitors, as well as calorie intake restriction and special diets. Most of the available results were insufficient and could not provide convincing evidence for the life span extension and the safety of the suggested geroprotectors. Drugs and means prolonging the life span could be subdivided into three groups: (a) geroprotectors prolonging the life span equally in all the members of the population: these postponed the beginning of the population's aging; (b) geroprotectors decreasing the mortality rate in a long lived subpopulation, which raised their maximal life span: these slowed down the population's aging rate; (c) geroprotectors increasing the survival rate in a short-lived subpopulation without changes in the maximal life span: in this case, the aging rate increased. There was a high positive correlation between the type of geroprotector-induced aging delay and the pattern of tumour development in the same population of animals. The first type of geroprotectors did not influence the incidence of tumour but increased tumour latency. The second type of geroprotectors was effective both in the inhibition of spontaneous carcinogenesis and the increase in tumour latency. Certain drugs of the third type raised tumour incidence in the exposed populations. According to the multistage model, geroprotectors either inhibit or accelerate the passage of carcinogen-exposed cells form one stage to another. Thus, the efficacy of geroprotectors as preventive means of cancer development will decrease with respect to the age of exposure onset. Recommendations of the available drugs and means of life span increase should be carefully reconsidered under the international scientific control. PMID- 11404055 TI - Locomotor activity as a function of age and life span in Drosophila melanogaster overexpressing hsp70. AB - Heat shock protein induction might be responsible for the longevity increase conferred by exposure to non-lethal stresses. To test this hypothesis, we studied in transgenic Drosophila melanogaster overexpressing hsp70 and controls, two behavioral variables (spontaneous locomotor activity and climbing activity) to evaluate the rate of aging, and life span. The results showed that in flies kept in groups, life span was decreased in transgenic flies compared to the parental line, but the contrary was observed in individually kept flies. Hsp70 overexpression had no dramatic effect on life span. Furthermore, we did not detect any advantage of Drosophila overexpressing hsp70 on the two measurements of locomotor activity. These results indicate that the rate of aging in transgenic flies is not different than in non-transgenic lines and that they are not more able to cope with the effects of aging on locomotor activity. PMID- 11404056 TI - Lipid peroxidation in the heart of adult and old rats during immobilization stress. AB - This study was designed to assay the lipid peroxidation (LPO) in the heart from adult (10--12 months) and old (22--25 months) Wistar male rats during immobilization stress. A value of LPO was estimated according to the concentration of conjugate dienes, TBA-positive substances and fluorescent products such as Schiff base. Investigations have shown that immobilization of rats was accompanied by increase in the concentration of all studied LPO intermediates in myocardium from both the age groups. Accumulation of these metabolites in old rat hearts was considerably higher than in adult ones. Dimethyl sulfoxide injection has limited stress-induced stimulation of LPO. Changes revealed in LPO-stimulation have different consequences in pathogenesis of stress-dependent injury in myocardium from adult and old rats. Utilization of LPO metabolites in old rats takes place mainly with formation of Schiff base. Accumulation of these products in myocardium is accompanied by modification of intracellular macromolecules and thus promotes the enhancement of stress-induced injury of myocardial cells. In adult rats during stress, the utilization of malonic dialdehyde and substances homologous to it is not accompanied by Schiff base accumulation in myocardium. It may be important for the restriction of stress-induced injury of myocardial cells. PMID- 11404057 TI - Mitochondrial DNA polymorphism: its role in longevity of the Irish population. AB - The mtDNA genome has been implicated as playing a pivotal role in determining the longevity and success of the human lifespan. A PCR-RFLP methodology was used to identify polymorphic restriction enzyme sites within a 2643 bp region of the mtDNA genome and a table of genetic haplotypes for a healthy aged and a younger control cohort of patients was constructed. Forty-six different mtDNA haplotypes and 11 groups of related haplotypes were identified across the two age groups but statistical analysis failed to show any significant associations. The European J haplogroup, previously reported to be associated with longevity, was not found at an increased frequency within the Irish aged population (P=0.36). However, the haplotypes comprising the J haplogroup could be differentiated into two distinct branches by the presence or absence of the two polymorphic restriction sites, 16,389g and 16,000g. The branch of haplotypes defined by 16,389g displayed a significant increased frequency in the aged samples (8%) compared to the controls (1%), P=0.015. Inversely, the branch of haplotypes defined by 16,000g displayed a significant decreased frequency in the aged samples (4%) compared to the controls (13%), P=0.011. The polymorphism (mt5178A) associated with longevity in the Japanese was not found in the Irish population, while the polymorphism (mt9055A) associated with successful ageing in the French centenarians was found at an increased frequency in the Irish aged population (9%) compared to the younger control group (5%), but failed to reach a level of statistical significance, P=0.164. PMID- 11404058 TI - Induction of complement C9 messenger RNAs in human neuronal cells by inflammatory stimuli: relevance to neurodegenerative disorders. AB - Neurons express proteins of the classical complement pathway, including C9. Both the mRNA and protein levels for C9 are sharply upregulated in brain areas affected by Alzheimer's disease (AD). Since little is known about the signals that are responsible for this upregulation, we evaluated in human SH-SY5Y neuroblastoma cells the factors which stimulate C9 production. Interferon-gamma, phorbol myristate acetate and interleukin-6 all stimulated C9 mRNA expression but the inflammatory cytokines tumor necrosis factor-alpha, interleukin-1 beta, as well as the anaphylatoxin C5a and the bacterial lipopolysaccharide, were ineffective. Immunohistochemical analysis of postmortem human brains for C9 protein demonstrated its presence in many cortical pyramidal neurons in AD, Down's syndrome, the parkinsonism dementia complex of Guam and pallido-ponto nigral degeneration, as well as in thalamic neurons of progressive supranuclear palsy and ballooned neurons of Pick's disease. Since C9 is required for the membrane attack complex of complement to become functional, interfering with signaling pathways that stimulate its production could offer new therapeutic strategies for treating various neurodegenerative disorders. PMID- 11404059 TI - DNA-microarrays: novel techniques to study aging and guide gerontologic medicine. AB - DNA array technology is a high throughput application of nucleic acid hybridization. Arrays currently used include spotted cDNA arrays at various densities and oligonucleotide arrays, where the highest densities can be obtained by synthesizing oligonucleotides directly on the solid support. While cDNA arrays can only determine the presence of a specific sequence in a sample, oligonucleotide arrays additionally allow detection of mutations and polymorphisms, including single nucleotide polymorphisms. Microarray technology can be expected to further our understanding of the aging process and to soon influence medical decision-making in age-related diseases, as well as lifestyle decisions of consequence to the aging process. PMID- 11404060 TI - Digital light microscopy: prerequisite for optimum contrast enhancement and increase of resolution. AB - Since the reproducible production of microscope objectives was enabled by the lens calculations of Ernst Abbe in 1872, various attempts have been made to further increase the resolution of light microscopes. Apart from the improvements on the optical side, especially the introduction of fluorescence methods, the use of digital cameras connected to computers have brought us close to the theoretical limits of optical resolution. Due to improved speed and memory capacity of modern computers mathematical methods can be applied to stored three dimensional (3D) sequences of digital images which, in addition to just contrast and edge enhancement, may result in the case of real 3D deconvolution, even in the visualisation of structures beyond the theoretical limitations of light optical resolution. PMID- 11404061 TI - Comparison of PCR and common clinical tests for the diagnosis of H. pylori in dyspeptic patients. AB - Helicobacter pylori has been recognized as a major gastric pathogen. The objective of this study was to assess the diagnostic value of common clinical tests to detect H. pylori infection, by comparison with PCR. Serum and gastric biopsy specimens from 106 dyspeptic patients were examined. Serology was performed with Pyloriset Dry test, and biopsies were examined histologically, for rapid urease activity and PCR amplification of an ureA gene segment of H. pylori. PCR primers were specific for H. pylori and required at least 1.47 pg of H. pylori DNA, corresponding to about 800 bacterial cells. According to serology, histology, rapid urease, and PCR, positive results were respectively found in 56%, 86%, 64%, and 85% of dyspeptic patients, primarily with gastritis. Relative to PCR, the sensitivity (and specificity) was 55% (38%) for serology, 86% (13%) for histology, 70% (69%) for urease. When combining histology and urease, Bayesian analysis of data indicated no advantage of using combined methods over rapid urease test alone. Histology should not any longer be considered a gold standard test for Helicobacter pylori. Urea breath test still seems the first option for non invasive diagnostic. If an invasive diagnostic is justified, highly specific and sensitive molecular methods should be used to examine specimens. PMID- 11404062 TI - Effect of sputum induction by hypertonic saline on specimen quality. AB - Microbiologic work-up of expectorated material is routinely used to search for the etiology of pulmonary infections, but sputum is often contaminated by saliva. Inhalation of hypertonic saline induces bronchial secretions and theoretically may improve specimen quality. We compared in a laboratory-blinded, randomized study the quality of sputum obtained either with induction by saline or without induction in patients with respiratory tract infection and a history of productive cough. The quality of sputum was considered good if the polymorphonuclear neutrophils (PMNs)/squamous epithelial cells (SECs) ratio was > or = 2 or the Q (quality) score was > or = +1 on Gram stain. Forty-nine and 50 patients were able to expectorate in the induced and spontaneous sputum groups, respectively. PMNs/SECs ratio was > or = 2 in 65% and 74% of the patients in the induced and spontaneous sputum groups (p = 0.47); for the Q score, a value > or = +1 was found in 55% and 66% of the patients of each group (p = 0.37). In conclusion, sputum induction by hypertonic saline inhalation does not improve specimen quality as judged by the PMNs/SECs ratio on Gram stain. PMID- 11404063 TI - Efficacy of two DNA fingerprinting methods for typing Acinetobacter baumannii isolates. AB - Performance of macrorestriction and repetitive extragenic palindromic DNA sequence-based PCR (REP-PCR) to type Acinetobacter baumannii isolates was quantitatively estimated using a test population of 54 outbreak-related, 29 endemic infection-related and 17 epidemiologically-unrelated isolates. Reproducibility and stability for macrorestriction were 100%, and REP-PCR showed only slightly lower stability. Macrorestriction resolved 18 fingerprints and REP PCR 10 DNA patterns, forming eight and seven clusters at 75% of similarity level, respectively. Intercluster band variation was > 7 bands for both methods. Although, all endemic isolates, except one, were concordantly grouped by both methods, macrorestriction distinguished a greater number of subtypes over one year study. For outbreaks, the epidemiologic concordance for both methods was 88%. The discriminatory index for macrorestriction and REP-PCR was 0.884 and 0.877, respectively. In conclusion, both methods showed similar efficacy as epidemiological markers, and by concordance, this study demonstrated that for REP PCR typing, a > or = 7 bands difference seemed an appropriate threshold to identify unrelated strains. PMID- 11404064 TI - A newer approach for the laboratory diagnosis of tuberculous meningitis. AB - In this prospective study, a simple method was standardized for measuring circulating mycobacterial antigen in the cerebrospinal fluid (CSF) for the laboratory diagnosis of tuberculous meningitis (TBM). The heat-inactivated CSF specimens from tuberculous and non-tuberculous patients were subjected to sodium dodecyl sulfate (SDS) - polyacrylamide gel electrophoresis (PAGE) (SDS-PAGE) and they were subsequently transferred onto nitrocellulose membrane (NCM) Using a rabbit polyvalent antibody to M tuberculosis, a heat stable 82 kDa mycobacterial antigen was demonstrated in the CSFs of patients with TBM. This antigen was conspicuous by its absence in the CSFs of non-tuberculous subjects. Due to inactivation of CSF specimens, there is a minimal risk of handling of infectious material in the laboratory. Besides, this newer approach is simple, inexpensive and can be readily applied in any routine clinical laboratory and it is particularly suited to developing countries. PMID- 11404065 TI - Comparison of MB/BacT system and agar proportion method in drug susceptibility testing of Mycobacterium tuberculosis. AB - The drug susceptibilities of 105 isolates/strains of Mycobacterium tuberculosis (101 clinical isolates and four control strains from the American Type Culture Collection) were assessed by the MB/BacT system and conventional agar proportion method. The agreement rates between the two methods were 99.0% for streptomycin, 95.2% for isoniazid and 100% for rifampin. The mean times to detection for drug resistant isolates were 4.7 days (range: 2.5-13.7 days) using the MB/BacT system and 14.8 days (range: 14-21 days) using the agar proportion method. For drug susceptible isolates, the times to detection were 10.8 days (mean) and 21 days respectively. Thus, these data have demonstrated that the automated, non radiometric MB/BacT system is an efficient, accurate and reliable method for assessing drug susceptibilities of M. tuberculosis compared with the conventional agar proportion method. PMID- 11404066 TI - Use of real-time PCR and the LightCycler system for the rapid detection of Pneumocystis carinii in respiratory specimens. AB - Pneumocystis carinii pneumonia (PCP) remains a major cause of morbidity and mortality in immunocompromised patients, including those infected with human immunodeficiency virus (HIV). The advent of real-time PCR technology offers the potential for rapid PCR results for the detection of P. carinii. In this report we describe the modification and evaluation of an existing PCR-based method for the detection of P. carinii DNA, into a real-time PCR assay suitable for use with the LightCycler system. Twenty eight induced sputum and bronchial washing specimens from 28 patients were tested by both a conventional PCR assay and a real-time PCR assay. Twelve specimens (42.9%) were positive in both the conventional and real-time PCR assays and sixteen (57.1%) were negative in both assays. The melting points of the amplified P. carinii DNA product obtained by melting curve analysis by the LightCycler of all P. carinii positive specimens ranged from 81.5 degrees C to 83.9 degrees C. There were no discordant results between the two assays for any of the specimens tested and results were available within 2 h for the real-time PCR assay compared to up to 11 h for the conventional PCR assay. PMID- 11404067 TI - Can antimicrobial susceptibility testing results for ciprofloxacin or levofloxacin predict susceptibility to a newer fluoroquinolone, gatifloxacin?: Report from The SENTRY Antimicrobial Surveillance Program (1997-99). AB - A serious problem confronting clinical laboratories and hospital formulary practices is the delayed availability of approved, commercially prepared susceptibility test reagents for newer antimicrobials. A current example is gatifloxacin, a new 8-methoxy fluoroquinolone with expanded potency against many Gram-positive pathogens. This study addresses the use of "surrogate marker" fluoroquinolones to predict susceptibility for gatifloxacin. Reference broth microdilution MIC results for 29,632 strains isolated in United States medical centers (SENTRY Antimicrobial Surveillance Program, 1997-99) were used: staphylococci (9,940 strains), enterococci (2,570), Streptococcus pneumoniae (3,784), Enterobacteriaceae (10,670) and Pseudomonas aeruginosa (2,668). Gatifloxacin interpretation categories were compared to those of ciprofloxacin and levofloxacin by regression statistics and error rate bounding analyses. For the Enterobacteriaceae, the absolute categorical agreement was 97.9 to 98.7% (false-susceptible or very-major error [VME], 0.03%-0.1%) for comparisons of both ciprofloxacin and levofloxacin with gatifloxacin. P. aeruginosa testing was more problematic (higher minor error rates), but acceptable at 0.6% to 1.1% VME and a 85.7% to 89.9% overall agreement. Ciprofloxacin results used to predict gatifloxacin in Gram-positive species was almost without VME (0.0%-0.2%) because gatifloxacin was significantly superior against these species, especially for S. pneumoniae, where gatifloxacin (MIC(90,) 0.5 microg/ml) was fourfold more potent than levofloxacin (MIC(90,) 2 microg/ml). The preferred gatifloxacin predictor drug was ciprofloxacin for all species except S. pneumoniae and P. aeruginosa, where levofloxacin results had a slightly greater predictive value. Susceptibility testing results for selected currently available fluoroquinolones can be used to predict susceptibility to gatifloxacin with high confidence. Many Gram-positive cocci, however, will be categorized as false-resistant by this interim method since gatifloxacin has a 11% to 34% wider spectrum of activity compared to ciprofloxacin when testing staphylococci and enterococci. Clinical laboratories can reliably use these suggested "surrogate markers" until reliable tests for gatifloxacin become available. PMID- 11404068 TI - Activity of BMS284756 against 2,681 recent clinical isolates of Haemophilus influenzae and Moraxella catarrhalis: Report from The SENTRY Antimicrobial Surveillance Program (2000) in Europe, Canada and the United States. AB - Although the isolation and detection of fluoroquinolone-resistant Haemophilus influenzae and Moraxella catarrhalis has been a very rare occurrence, newer agents in the quinolone class must be evaluated to determine their comparative potencies and to develop in vitro testing methods. BMS284756 is an investigational desfluoro(6)-quinolone with a spectrum of activity most similar to recently introduced agents such as gatifloxacin and trovafloxacin. This compound was compared to levofloxacin, gatifloxacin, ciprofloxacin and moxifloxacin, as well as other orally administered antimicrobials against 1,872 H. influenzae and 810 M. catarrhalis isolates. Two Canadian H. influenzae strains had ciprofloxacin MICs elevated above the normal wild type susceptible MIC population (> 0.06 microg/mL). All other strains of H. influenzae and M catarrhalis were highly susceptible to the tested quinolones (MIC(90,) < or = 0.016 or 0.03 microg/mL). For the two H. influenzae isolates with elevated quinolone MICs the potency rank order was: gatifloxacin and BMS284756 (MICs, 0.25 and 1 microg/mL) > levofloxacin and moxifloxacin > ciprofloxacin (MICs, 0.5 and > 2 microg/mL). The comparison of Etest (AB BIODISK, Solna, Sweden) and disk diffusion results to the reference broth microdilution values produced acceptable intermethod accuracy when applied to BMS284756. This novel desfluoro compound possesses promising activity against fastidious Gram-negative respiratory tract pathogens and further clinical development is underway. PMID- 11404069 TI - Effect of removal of duplicate isolates on cumulative susceptibility reports. AB - The objective of our study is to assess the impact of different methods of duplicate isolate removal on cumulative susceptibility reports. Over a 1-year period, we studied the effect of 3 methods of duplicate isolate removal on the cumulative percentage susceptibility of 9 Gram-negative bacilli to 15 antimicrobials. Raw data from which no duplicate isolates were removed (NR) were generated by the Sensititre breakpoint susceptibility testing system. D3 and D7 were methods of duplicate isolate removal defined as follows: same patient, bacterial species, irrespective of susceptibility within either three (D3) or seven (D7) calendar days of the date of the previous culture. The third method evaluated was an algorithm utilized by Cerner, a laboratory management program that defines duplicate isolates as follows: same patient, bacterial species, and NCCLS susceptibility category to an individual antimicrobial. Differences in percentage susceptibility between the three methods of duplicate isolate removal and NR were assessed. The number of isolates studied ranged from 80 (E. aerogenes) to 681 (P. aeruginosa). Of the methods of duplicate isolate removal, the highest percentage susceptibility occurred most frequently with Cerner followed by D7 and D3. Differences in percentage susceptibility between methods of removal and NR ranged from -11 to 25%, -5 to 8%, and -3 to 10%, with Cerner, D3, and D7, respectively. The percentage susceptibility was at least 5% higher than NR with a method of removal for 15 individual organism/antimicrobial combinations in which susceptibility was > or = 70% by at least one of the methods. These occurred most frequently with Enterobacter species and Cerner. Although there is no consensus on the ideal method of duplicate isolate removal, one should be cognizant that these manipulations may produce different cumulative susceptibility reports. PMID- 11404070 TI - Comparison of two commercial systems (Vitek and MicroScan-WalkAway) for antimicrobial susceptibility testing of Pseudomonas aeruginosa isolates from cystic fibrosis patients. AB - Antimicrobial susceptibility testing of cystic fibrosis (CF) isolates of Pseudomonas aeruginosa is difficult because the organisms are often mucoid and slow-growing. This study of 498 CF strains examined the correlation of results derived from two commonly used commercial systems (Vitek, MicroScan-WalkAway) with a reference method for 10 antimicrobials. Correlation to reference results was unacceptably low for all agents and both commercial systems had a high rate of very major (false-susceptible) errors. Although mucoid strains produced a 4.8% greater intermethod error, it was not markedly different than non-mucoid strains for the Vitek System. Overall, these tested commercial systems performed poorly for CF isolates in contrast to earlier reported, high correlations with the reference methods (broth microdilution frozen panels and agar dilution) of the National Committee for Clinical Laboratory Standards, the standardized disk diffusion test, and the Etest (AB BIODISK, Solna, Sweden). PMID- 11404071 TI - A case of splenic abscess due to Chlamydia pneumoniae. AB - In this report, a case of chlamydial disease with splenic abscess associated with Chlamydia pneumoniae antigen and antibody was described. On spleen biopsy of the patient, an antigen specific to C.pneumoniae was detected by immunofluorescence staining with a monoclonal antibody. Serologic studies revealed a high antibody titer to C.pneumoniae in sera collected from the patient and her husband. Treatment with the antibiotic minocycline improved her condition. PMID- 11404072 TI - Neisseria elongata subsp. elongata, as a cause of human endocarditis. AB - Neisseria elongata subsp. elongata, previously considered nonpathogenic, is a potential agent of human endocarditis. We report the second case of human endocarditis caused by this organism. The patient was successfully treated with Ceftriaxone alone for a total of six weeks. PMID- 11404073 TI - Accuracy of broth microdilution and E test methods for detecting chloramphenicol acetyl transferase mediated resistance in Streptococcus pneumoniae: Geographic variations in the prevalence of resistance in The SENTRY Antimicrobial Surveillance Program (1999). AB - High antimicrobial resistance rates in Streptococcus pneumoniae has caused a need for alternative therapies. Chloramphenicol is currently being reconsidered as an empiric treatment for respiratory tract infections particularly in developing countries. In this study, we assessed the ability of the reference broth microdilution and Etest (AB BIODISK, Solna, Sweden) methods to detect chloramphenicol resistance among pneumococci as compared to the chloramphenicol acetyltransferase (CAT) assay. In the 1999 SENTRY Antimicrobial Surveillance Program, 1671 S. pneumoniae strains from respiratory tract infections were collected from 49 participants located in the Americas and Europe. The rates of penicillin and macrolide non-susceptibility were 15.6-41.3 and 12.4-26.8%, respectively. All chloramphenicol-resistant strains were CAT assay positive (n = 154; 9.2% of isolates) with highest resistance rates in Europe (12.7%; range among sites, 0.0-38.5%) and the United States (10.6%; range, 0.0-25.6%). Etest MICs correlated with reference results and the current breakpoint for chloramphenicol resistance (> or = 8 microg/mL) remains valid for S. pneumoniae and Haemophilus influenzae (eight strains tested). CAT-mediated resistances dominate among chloramphenicol-resistant S. pneumoniae, and marked geographic variations in susceptibility were discovered. PMID- 11404074 TI - Absence of mutation in the region (nt. 710-1010) of pbp4 gene in clinical isolates of Staphylococcus aureus with low-level teicoplanin resistance. AB - Resistance to teicoplanin in clinical isolates of Staphylococcus aureus has increased in frequency. Detection is difficult due to heterogeneous phenotypes of these strains. Two mutations in pbp4 have recently been identified in glycopeptide resistant laboratory strains. We investigated to determine if these mutations could be used to screen clinical isolates for teicoplanin resistance. Of 41 clinical isolates screened, none contained the mutations in pbp4 indicating these mutations cannot be used as a molecular diagnostic tool for glycopeptide resistance. PMID- 11404076 TI - The origin of the ankyrin repeat region in Notch intracellular domains is critical for regulation of HES promoter activity. AB - Notch signal transduction is mediated by proteolysis of the receptor and translocation of the intracellular domain (IC) into the nucleus, where it functions as a regulator of HES gene expression after binding to the DNA-binding protein RBP-J kappa. The mammalian Notch receptors are structurally very similar, but have distinct functions. Most notably, Notch 1 IC is a potent activator of the HES promoter, while Notch 3 IC is a much weaker activator and can repress Notch 1 IC-mediated HES activation in certain contexts. In this report we explore the molecular basis for this functional difference between Notch 1 and Notch 3 IC. We find that Notch 3 IC, like Notch 1 IC, can bind the SKIP and PCAF proteins. Furthermore, both Notch 1 and Notch 3 ICs displace the co-repressor SMRT from the DNA-binding protein RBP-J kappa on the HES promoter. The latter observation suggests that both Notch 3 IC and Notch 1 IC can access RBP-J kappa in vivo, and that the difference in activation capacity instead stems from structural differences in the two ICs when positioned on RBP-J kappa. We show that two distinct regions in the Notch IC are critical for the difference between the Notch 1 and Notch 3 IC. First, the origin of the ankyrin repeat region is important, i.e. only chimeric ICs containing a Notch 1-derived ankyrin repeat region are potent activators. Second, we identify a novel important region in the Notch IC. This region, named the RE/AC region (for repression/activation), is located immediately C-terminal to the ankyrin repeat region, and is required for Notch 1 IC's ability to activate and for Notch 3 IC's ability to repress a HES promoter. The interplay between the RE/AC region and the ankyrin repeat region provides a basis to understand the difference in HES activation between structurally similar Notch receptors. PMID- 11404077 TI - Early anteroposterior division of the presumptive neurectoderm in Xenopus. AB - We analyze the timing of neural patterning in Xenopus and the mechanism by which the early pattern is generated. With regard to timing, we show that by early gastrula, two domains of the anteroposterior (A/P) pattern exist in the presumptive neurectoderm, since the opl gene is expressed throughout the future neural plate, while the fkh5 gene is expressed only in more posterior ectoderm. By mid-gastrula, this pattern has become more elaborate, with an anterior domain defined by expression of opl and otx2, a middle domain defined by expression of opl and fkh5, and a posterior domain defined by expression of opl, fkh5 and HoxD1. Explant assays indicate that the late blastula dorsal ectoderm is specified as the anterior domain, but is not yet specified as middle or posterior domains. With regard to the mechanism by which the A/P pattern is generated, gain and loss of function assays indicate that quantitatively and qualitatively different factors may be involved in inducing the early A/P neural pattern. These data show that neural patterning occurs early in Xenopus and suggest a molecular basis for initiating this pattern. PMID- 11404078 TI - A gene encoding a new ONECUT class homeodomain protein in the ascidian Halocynthia roretzi functions in the differentiation and specification of neural cells in ascidian embryogenesis. AB - Genes encoding a novel group of homeodomain transcription factors, ONECUT class homeodomain proteins, have previously been isolated from vertebrate and insect. Among them, vertebrate HNF-6 is expressed in hepatocytes and the central nervous system during embryogenesis. Although the functions of HNF-6 in hepatocytes have been well studied, the functions of HNF-6 in the central nervous system remain unknown. In this study, we isolated HrHNF-6, which encodes a new ONECUT class homeodomain protein, from an ascidian, Halocynthia roretzi. HrHNF-6 mRNA was expressed exclusively in neural cells, just posterior to the expression of Hroth during embryogenesis. One of the functions of HrHNF-6 in neural cells is the activation of the expression of HrTBB2, the ascidian beta-tubulin gene. Another is the restriction of the expression of HrPax-258 (which is expressed in the neural tube), suggesting that HrHNF-6 functions in the specification of the neural tube. These results indicate that HrHNF-6 functions in the differentiation and regional specification of neural cells during ascidian embryogenesis. PMID- 11404079 TI - A role for the COUP-TF-related gene seven-up in the diversification of cardioblast identities in the dorsal vessel of Drosophila. AB - The Drosophila gene tinman is essential for dorsal vessel (heart) formation and is structurally and functionally conserved in vertebrates. In the mature embryonic dorsal vessel, tinman is expressed in four of the six pairs of cardioblasts in each segment. We provide evidence that seven-up, which is homologous to the vertebrate COUP-TF transcription factor and is expressed in the non-Tinman-expressing cardioblasts, represses tinman in these cells. Loss of function seven-up mutations derepress tinman expression in these cardioblasts while ectopic expression of seven-up represses tinman in the cardioblasts that normally express it. These changes are correlated with alterations in the expression of additional molecular markers for each of these two types of cardioblasts, such as the novel T-box-containing gene Tb66F2 and the potassium channel-encoding gene sur. These observations suggest that seven-up has a role in diversifying cardioblast identities within each segment. We also describe the tinman cis sequences that mediate tinman repression by seven-up and examine whether Seven-up can bind these sequences to directly inhibit tinman. PMID- 11404080 TI - Smad5 is required for mouse primordial germ cell development. AB - Smad5, together with Smad1 and Smad8, have been implicated as downstream signal mediators for several bone morphogenetic proteins (BMPs). Recent studies have shown that primordial germ cells (PGCs) are absent or greatly reduced in Bmp4 or Bmp8b mutant mice. To define the role of Smad5 in PGC development, we examined PGC number in Smad5 mutant mice by Oct4 whole-mount in situ hybridization and alkaline phosphatase staining. We found ectopic PGC-like cells in the amnion of some Smad5 mutant mice, however, the total number of PGCs was greatly reduced or completely absent in Smad5 mutant embryos, similar to Bmp4 or Bmp8b mutant embryos. Therefore, Smad5 is an important factor involved in PGC generation and localization. PMID- 11404081 TI - Cellular mechanisms in the development of the Drosophila arista. AB - Epidermal cells of Drosophila form a variety of polarized structures during their differentiation. These polarized structures include epidermal hairs, the shafts of sensory bristles, larval denticles and the arista laterals. The arista is the terminal segment of the antenna and consists of a central core and a series of lateral extensions. Here we describe the cellular mechanisms involved in the development of the arista and the morphogenesis of the laterals. We found that the development of the arista is a complex process that involves coordinated cell shape changes, elongation of the central core, apoptosis, nuclear migration, the formation of polyploid cells and the outgrowth of the laterals. This developmental program is highly conserved in the development of the arista in the housefly (Musca domestica). Altering arista cell number in Drosophila by stimulating or inhibiting apoptosis results in an altered number of laterals. Interestingly, the increased number of laterals that result from the inhibition of apoptosis in Drosophila results in an arista whose morphology is reminiscent of the Musca arista. Previous experiments have shown that both the actin and microtubule cytoskeletons have important functions in the cellular morphogenesis of hairs and bristles. Inhibitor studies reported here show that this is also the case for the formation of the arista laterals, arguing that the actin and microtubule cytoskeletons have similar functions in the morphogenesis of all of these cell types. We conclude that the arista laterals are a valuable complementary cell type system for studying the morphogenesis of polarized cellular extensions in Drosophila. PMID- 11404082 TI - Heterogeneous nuclear ribonucleoprotein-H plays a suppressive role in visceral myogenesis. AB - Mouse embryonic mesenchymal cells undergo spontaneous smooth muscle (SM) differentiation upon spreading/elongation in culture (Relan et al., J. Cell Biol. 147 (1999) 1341; Yang et al., Development 125 (1998) 2621; Yang et al., Development 126 (1999) 3027). Using these cells we generated a subtracted cDNA library to identify potential suppressors of SM myogenesis. One of the differentially expressed genes was heterogeneous nuclear ribonucleoprotein-H (hnRNP-H), which is involved in pre-mRNA alternative splicing. hnRNP-H was highly expressed in mesenchymal cells prior to the onset of SM differentiation, but its expression rapidly decreased in mesenchymal cells undergoing SM myogenesis. In vivo, the drop in hnRNP-H expression was restricted to visceral SM cells. Antisense oligodeoxynucleotide and antisense RNA were used to inhibit hnRNP-H synthesis in SM-differentiating mesenchymal cells and in embryonic lung explants. A decrease in hnRNP-H levels resulted in upregulation of SM-specific gene expression and increased bronchial SM development in lung explants. hnRNP-H overexpression in cell cultures had the opposite effect. These studies, therefore, indicate a novel role for hnRNP-H in the control of visceral myogenesis. PMID- 11404083 TI - Mouse mammary gland involution is associated with cytochrome c release and caspase activation. AB - At weaning, milk producing mammary epithelial cells undergo apoptosis and are removed by phagocytosis. Here, we show that mouse mammary gland involution is associated with mitochondrial cytochrome c release and processing of numerous caspases, including caspase-1, -3, -7, -8 and -9. Induction of caspase-3-like activity paralleled cleavage of poly-(ADP--ribose) polymerase. Dexamethasone inhibited processing of caspase-3, -7 and -8 and apoptosis, but had no effect on caspase-1 accumulation and cytochrome c release. In Bcl-2 transgenic animals, cytochrome c release, caspase activation and apoptosis were impaired. Thus, the pro-apoptotic signaling pathway in mammary epithelial cells during involution involves the release of cytochrome c and activation of caspases. It is inhibited by Bcl-2 at the mitochondrial level and by dexamethasone at a post-mitochondrial level. PMID- 11404084 TI - New members of the Drosophila Myc transcription factor subfamily revealed by a genome-wide examination for basic helix-loop-helix genes. AB - The basic helix-loop-helix (bHLH) proteins represent an evolutionary conserved class of transcription factors that are known to play important roles in cell determination and differentiation during animal embryonic development. Following an exhaustive search of the complete Drosophila genome sequence using a PSI-BLAST strategy, we identified 19 new genes, bringing the total number of bHLH-encoding genes in the Drosophila genome to 56. These new genes belong to various subfamilies of bHLH transcription factors, such as the Daughterless, Hairy Enhancer of split, bHLH-PAS or bHLHZip subfamilies. The embryonic expression pattern of each of these new genes has been analyzed by in situ hybridization. By looking for closely structurally related motifs, we found two genes that represent likely orthologues of vertebrate Mnt and Mlx. Together with previous reports, our data suggest that, similar to networks involved in neurogenesis and myogenesis, the network of Myc-related genes has been globally conserved throughout evolution. PMID- 11404085 TI - Hemogen is a novel nuclear factor specifically expressed in mouse hematopoietic development and its human homologue EDAG maps to chromosome 9q22, a region containing breakpoints of hematological neoplasms. AB - We cloned a novel murine gene, designated Hemogen (hemopoietic gene), which was sequentially expressed in active hematopoietic sites and downregulated in the process of blood cell differentiation. Hemogen transcripts were specifically detected in blood islands, primitive blood cells and fetal liver during embryogenesis, and then remained in bone marrow and spleen in adult mice. Immunostaining demonstrated that Hemogen was a nuclear protein. We also identified a human homologue of Hemogen, named EDAG, which was mapped to chromosome 9q22, a leukemia breakpoint. Like Hemogen, EDAG exhibited specific expression in hematopoietic tissues and cells. Taken together, these data are consistent with Hemogen and EDAG playing an important role in hematopoietic development and neoplasms. PMID- 11404086 TI - Expression of zebrafish btg-b, an anti-proliferative cofactor, during early embryogenesis. AB - BTG/tob family proteins are thought to be a potential tumor suppressor due to their anti-proliferative activity. We cloned zebrafish btg-b, a member of the BTG1/2 subfamily, using in situ hybridization screening. The tissue-specific expression of btg-b is first observed in the organizer region at the early gastrula stage. Later in development, the forebrain, the hindbrain, the polster and the paraxial mesoderm transiently express btg-b. Recently, mouse Btg1 and Btg2 have been shown to be a cofactor for Hoxb9. Double in situ hybridization with zebrafish btg-b and hoxb9a indicates that the expression domains of these two genes overlap in the posterior paraxial mesoderm. PMID- 11404087 TI - Cloning and expression of noz1, a zebrafish zinc finger gene related to Drosophila nocA. AB - We report the isolation of noz1, a novel zebrafish zinc finger gene which displays sequence similarity to Drosophila nocA. noz1 transcripts are detected at the shield stage within the germ ring and excluded from the most dorsal region. By the end of gastrulation, noz1 is expressed in the presumptive hindbrain and spinal cord as well as in the forming tailbud. During somitogenesis noz1 shows a dynamic expression in the midbrain-hindbrain boundary, hindbrain and spinal cord. This results, at 24 hpf, in a graded expression with the highest level in rhombomeres 2 and 3, and the lowest in the spinal cord. Expression analysis in swirl and chordino mutants as well as in retinoic acid treated embryos indicate that noz1 is activated by BMP antagonists and neural posteriorizing signals. PMID- 11404088 TI - Isolation of the avian homologue of the homeobox gene Mox2 and analysis of its expression pattern in developing somites and limbs. AB - We have isolated the cDNA of avian Mox2 and analyzed its expression pattern during somitogenesis and limb bud formation. Mox2 plays an important role in limb muscle differentiation in the mouse. Mox2 is expressed in the somites of developing chick embryos and in presumptive migrating myoblasts from the dermomyotome to the limb buds. It is also expressed in the ventral and dorsal part of limb buds and is associated with non-proliferating myoblasts. Significant differences were observed in chick and mouse expression patterns, namely in the chick dermomyotome and limb. PMID- 11404089 TI - The expression pattern of tomoregulin-1 in urodele limb regeneration and mouse limb development. AB - Tomoregulin-1 (TMEFF1) was first identified as a gene implicated in pituitary secretion in Xenopus laevis. The predicted structure of TMEFF1 is that of a transmembrane protein with a highly conserved cytoplasmic tail, two follistatin domains and one modified EGF domain in its extracellular region. We report the cloning of the newt orthologue, and show that the expression of TMEFF1 is upregulated in the blastema during limb regeneration, and is also expressed in mouse embryonic limb development. PMID- 11404090 TI - Expression of the receptor tyrosine kinase gene EphB3 during early stages of chick embryo development. AB - The expression pattern of the receptor tyrosine kinase gene EphB3 was examined during the early stages of chick embryogenesis, and is described in this report. In the gastrula, EphB3 is expressed in epiblast cells adjacent to and entering the anterior portion of the primitive streak; expression is extinguished once cells have ingressed. At headfold stages, EphB3 is strongly transcribed in the floor of the foregut and in anterior lateral endoderm, and is expressed in the subjacent cardiogenic mesoderm. EphB3 is transiently expressed in the lateral ectoderm, neural tube, and neural crest during these stages. Later neural expression is localized to the mesencephalon. In the somitic mesoderm, EphB3 is initially expressed in the sclerotome, but later is expressed predominantly in the dermatome. Prominent expression is also detected in the developing heart, liver, posterior ventral limb bud mesenchyme, pharyngeal arches, and head mesenchyme. PMID- 11404091 TI - Comparative analysis of the expression patterns of Wnts and Frizzleds during early myogenesis in chick embryos. AB - During embryogenesis, the transduction of Wnt signals through Frizzled receptors is thought to play integral roles in myogenesis and somite patterning. However, little is known about which Wnt-Frizzled interactions are required for skeletal myogenesis. Thus, we sought to determine which Wnts and Frizzled exhibit expression patterns that are spatiotemporally consistent with the expression of two myogenic determination factors: Myf-5 and MyoD. To accomplish this, we first isolated partial cDNAs for six chick Frizzled orthologues and then compared the expression patterns of chick Frizzleds and Wnts to myogenic and somite patterning factors, such as Myf-5, MyoD, Sonic Hedgehog (Shh), Pax-1 and Pax-3 in Hamburger and Hamilton stage 10 chick. We used these data to generate a schematic composite of expression patterns at the level of the segmental plate and developing somites (stage V) that shows multiple Frizzled and Wnt transcripts expressed in tissues that are overlapping and adjacent to Myf-5 and MyoD expressing tissues. PMID- 11404092 TI - Desrt, an AT-rich interaction domain family transcription factor gene, is an early marker for nephrogenic mesoderm and is expressed dynamically during mouse limb development. AB - Desrt is a mouse gene of the AT-rich interaction domain family of transcription factors. Here we describe the temporal and spatial pattern of expression of Desrt during mouse organogenesis. Desrt expression is first detected in the intermediate plate mesoderm, providing an early embryonic marker for this tissue, and subsequently in the nephrogenic cords of the urogenital ridges. A highly dynamic expression pattern is observed in the developing limb, implicating Desrt in limb patterning. Desrt is also detected in the myotome of the somites, the oro naso-pharyngeal ectoderm and underlying mesenchyme, otic vesicles, the gut and its derivatives, and transiently in the liver. PMID- 11404093 TI - Embryonic expression of the murine homologue of SALL1, the gene mutated in Townes -Brocks syndrome. AB - SALL1 is one of three human homologues of the Drosophila region-specific homeotic gene spalt (sal). Mutations of SALL1 on chromosome 16q12.1 cause Townes--Brocks syndrome (TBS) which is characterized by defects in multiple organ systems including limbs, ears, kidneys and anus. Here, we have analyzed the expression of the mouse homologue of SALL1 (Sall1) during early embryogenesis. Sall1 expression is very prominent in the developing brain and the limbs. Other sites of expression include the meso- and metanephros, lens, olfactory bulbs, heart, primitive streak and the genital tubercle. Hence, Sall1 expression to a large degree reflects the structures affected in human TBS. PMID- 11404094 TI - Xenopus Smad3 is specifically expressed in the chordoneural hinge, notochord and in the endocardium of the developing heart. AB - The Smads are intracellular signalling molecules that transduce signals from receptors for members of the TGF-beta superfamily to the nucleus. We have cloned the Xenopus orthologue of Smad3 (XSmad3). It is 94.6% identical to human Smad3 at the amino acid level. It is expressed as a maternal mRNA which disappears after stage 10.5, but reappears at the early tailbud stages. It is much less abundant than XSmad2 at the early developmental stages. From Stage 27 onwards XSmad3 is expressed with XSmad2 throughout the head region and in the somitic region. Strikingly however, XSmad3 alone is specifically expressed in the chordoneural hinge, the notochord and in the developing heart. Closer analysis reveals that XSmad3 is specifically expressed in the endocardium but not in the myocardium or pericardium. The chordoneural hinge staining persists at least until stage 40 whereas the staining in the endocardium peaks at approximately stage 32/33. PMID- 11404095 TI - Cloning and expression pattern of a novel PEBP2 beta-binding protein (charged amino acid rich leucine zipper-1[Crl-1]) in the mouse. AB - PEBP2 beta/Cbf beta is the beta subunit of PEBP2/Cbf, which has been demonstrated to have important biological activities in hematopoiesis and osteogenesis. However, PEBP2 beta is ubiquitously expressed, suggesting that PEBP2 has other additionally important physiological activities. In an effort to elucidate other possible functions for PEBP2, we have isolated a novel gene that encodes a PEBP2 beta-interacting protein from a mouse cDNA library. We have called this gene Crl 1 for charged amino acid rich leucine zipper-1 (Crl-1) because it is rich in charged amino acids and contains a putative leucine zipper region. Expression studies in a 17.5 days post-coitum mouse embryo demonstrated Crl-1 expression mainly in the olfactory bulb and cerebral cortex. Post-natally, Crl-1 expression was additionally observed in the cerebellar cortex with strong expression in the hippocampus. These findings show that this novel PEBP2 beta-interacting protein is expressed mainly in subsets of neuronal cells, suggesting that Crl-1 plays some role in the developing mouse brain. PMID- 11404097 TI - Web alert. Environmental biotechnology. Regulatory affairs. AB - A selection of World Wide Web sites relevant to papers published in this issue of Current Opinion in Biotechnology. PMID- 11404099 TI - The identification of microorganisms by fluorescence in situ hybridisation. AB - Fluorescence in situ hybridisation (FISH) with rRNA-targeted oligonucleotide probes facilitates the rapid and specific identification of individual microbial cells in their natural environments. Over the past year there have been a number of methodological developments in this area and new applications of FISH in microbial ecology and biotechnology have been reported. PMID- 11404100 TI - Microorganisms relevant to bioremediation. AB - Naturally occurring microbial consortia have been utilized in a variety of bioremediation processes. Recent developments in molecular microbial ecology offer new tools that facilitate molecular analyses of microbial populations at contaminated and bioremediated sites. Information provided by such analyses aids in the evaluation of the effectiveness of bioremediation and the formulation of strategies that might accelerate bioremediation. PMID- 11404101 TI - Biodegradation of plastics. AB - Widespread studies on the biodegradation of plastics have been carried out in order to overcome the environmental problems associated with synthetic plastic waste. Recent work has included studies of the distribution of synthetic polymer degrading microorganisms in the environment, the isolation of new microorganisms for biodegradation, the discovery of new degradation enzymes, and the cloning of genes for synthetic polymer-degrading enzymes. PMID- 11404102 TI - Microbial detoxification of metals and radionuclides. AB - Microorganisms have important roles in the biogeochemical cycling of toxic metals and radionuclides. Recent advances have been made in understanding metal-microbe interactions and new applications of these processes to the detoxification of metal and radionuclide contamination have been developed. PMID- 11404103 TI - Microbial dehalogenation. AB - Novel dehalogenases have been identified recently in various bacteria that utilise halogenated substrates. X-ray studies and sequence analysis have revealed insight into the molecular mechanisms of hydrolytic dehalogenases. Furthermore, genetic and biochemical studies have indicated that reductive dehalogenases are extra-cytoplasmic corrinoid-containing iron-sulphur proteins. Sequence analysis and mutagenesis studies indicate that several dehalogenases are homologous to enzymes that carry out transformations on non-halogenated substrates. PMID- 11404104 TI - Anaerobic biodegradation of saturated and aromatic hydrocarbons. AB - Saturated and aromatic hydrocarbons are wide-spread in our environment. These compounds exhibit low chemical reactivity and for many decades were thought to undergo biodegradation only in the presence of free oxygen. During the past decade, however, an increasing number of microorganisms have been detected that degrade hydrocarbons under strictly anoxic conditions. PMID- 11404105 TI - Recent advances in methane fermentation technology. AB - In the past two decades, a number of biotechnologies for anaerobic (methanogenic) wastewater treatment have been created, and practical applications of these processes are now being extended to more recalcitrant wastewaters and to wastewaters at extreme temperatures. Our knowledge of methanogenic organic degradation associated with bioreactors is also accumulating at a rapid rate. The recent advancement of such fundamental understanding is attributed to modern molecular biology techniques applied to the study of microbial communities and to continuous challenges to the cultivation of many important but recalcitrant anaerobes in bioreactors. PMID- 11404106 TI - Microbiology and application of the anaerobic ammonium oxidation ('anammox') process. AB - Ten years ago, an anaerobic ammonium oxidation ('anammox') process was discovered in a denitrifying pilot plant reactor. From this system, a highly enriched microbial community was obtained, dominated by a single deep-branching planctomycete, Candidatus Brocadia anammoxidans. Phylogenetic inventories of different wastewater treatment plants with anammox activity have suggested that at least two genera in Planctomycetales can catalyse the anammox process. Electron microscopy of the ultrastructure of B. anammoxidans has shown that several membrane-bounded compartments are present inside the cytoplasm. Hydroxylamine oxidoreductase, a key anammox enzyme, is found exclusively inside one of these compartments, tentatively named the 'anammoxosome'. PMID- 11404107 TI - Pseudomonas for biocontrol of phytopathogens: from functional genomics to commercial exploitation. AB - Pseudomonas spp. that can colonise the roots of crop plants and produce antifungal metabolites represent a real alternative to the application of chemical fungicides. Presently, much research is aimed at understanding, at the molecular level, the mechanisms that enable Pseudomonas strains to act as efficient biological control agents. This approach is facilitating the development of novel strains with modified traits for enhanced biocontrol efficacy. However, without solving some inherent problems associated with the effective delivery of microbial inoculants to seeds and without knowledge on the biosafety aspects of novel biocontrol agents, the commercial potential of Pseudomonas spp. for plant disease control will not be realised. PMID- 11404109 TI - The regulation of DNA vaccines. AB - The framework for regulating DNA vaccines has been in place since the first clinical trial was initiated in the mid-1990s. American and European regulatory guidance has evolved on the basis of insights provided by ongoing preclinical and clinical studies. These include analyses of the safety of DNA vaccines in normal volunteers, and recent data concerning the tissue distribution, persistence, and integration potential of DNA plasmids. PMID- 11404110 TI - The evolution of public review and oversight mechanisms in human gene transfer research: joint roles of the FDA and NIH. AB - The federal government is critically examining its responsibilities and opportunities for bringing the new field of gene therapy to fruition and for assuring public confidence in this new area of biomedicine. The evolving mechanisms for review and regulation in human gene transfer studies in the United States are being enhanced by increasingly effective interactions between the Food and Drug Administration and the National Institutes of Health. PMID- 11404111 TI - The regulation of biologic products derived from bioengineered plants. AB - Recently, there has been a large increase in the number and types of biological products--from therapeutic antibodies to vaccines for the prevention of infectious diseases--that are produced in bioengineered plant systems. We anticipate that this technology will be used increasingly on a commercial scale for the manufacture of human and animal products. These production systems have the capacity to produce very large quantities of products at lower costs and with reduced risks compared with mammalian systems. PMID- 11404112 TI - Xenotransplantation: regulatory challenges. AB - During 1999-2000, the US government published three xenotransplantation policy/guidance documents, one by the Public Health Service and two by the Food and Drug Administration (FDA). The FDA also held two public meetings of the xenotransplantation subcommittee of the Biological Response Modifiers Advisory Committee to discuss particular issues in xenotransplantation. PMID- 11404113 TI - Recent trends in cell substrate considerations for continuous cell lines. AB - Product development activity in the past five to ten years has reconstituted a version of an old debate on the safety assessment of biological products, namely whether the use of some types of continuous cell lines (CCLs) is appropriate in the preparation of some types of biological products. Since 1987, dozens of purified recombinant DNA products derived from CCLs have been developed and have received regulatory approval. In addition, several live attenuated and inactivated viral vaccines manufactured in CCLs were approved after thorough review of product safety and manufacturing issues. The current discussion revolves around the potential use of CCLs (human or not) to prepare purified protein subunit vaccines, such as for HIV, and the use of human CCLs to prepare purified protein products. PMID- 11404114 TI - Reference values of maximum isometric muscle force obtained in 270 children aged 4-16 years by hand-held dynamometry. AB - Since muscle force and functional ability are not related linearly; maximum force can be reduced while functional ability is still maintained. For diagnostic and therapeutic reasons loss of muscle force should be detected as early and accurately as possible. Because of growth factors, maximum muscle force in children varies with age, which makes detection of force loss difficult. The purpose of this study was to establish reference values for muscle force in children aged 4-16 years, obtained by hand-held dynamometry in 11 muscle groups. In boys muscle force was predicted best by weight whereas in girls weight and age were best predictors. At age 14 boys become significantly stronger for nearly all tested muscle groups. These age-related reference values can be used to quantify muscle weakness in individual muscle groups in children aged 4-16 years and to evaluate the effects of therapy. PMID- 11404115 TI - Rimmed vacuoles and the added value of SMI-31 staining in diagnosing sporadic inclusion body myositis. AB - Problems in diagnosing sporadic inclusion body myositis may arise if all clinical features fit a diagnosis of polymyositis, but the muscle biopsy shows some rimmed vacuoles. Recently, immunohistochemistry with an antibody directed against phosphorylated neurofilament (SMI-31) has been advocated as a diagnostic test for sporadic inclusion body myositis. The aims of the present study were to define a quantitative criterion to differentiate sporadic inclusion body myositis from polymyositis based on the detection of rimmed vacuoles in the haematoxylin-eosin staining and to evaluate the additional diagnostic value of the SMI-31 staining. Based on clinical criteria and creatine kinase levels in patients with endomysial infiltrates, 18 patients complied with the diagnosis of sporadic inclusion body myositis, and 17 with the diagnosis of polymyositis. A blinded observer counted the abnormal fibres in haematoxylin-eosin-stained sections and in SMI-31-stained sections. The optimal cut-off in the haematoxylin-eosin test was 0.3% vacuolated fibres. Adding the SMI-31 staining significantly increased the positive predictive value from 87 to 100%, but increased the negative predictive value only to small extent. We conclude that (1) patients with clinical and laboratory features of polymyositis, including response to treatment, may show rimmed vacuoles in their muscle biopsy and that (2) adding the SMI-31 stain can be helpful in differentiating patients who respond to treatment from patients who do not. PMID- 11404116 TI - Macrophagic myofasciitis associated with inclusion body myositis: a report of three cases. AB - We describe three patients with macrophagic myofasciitis and inclusion body myositis. All patients fulfilled diagnostic criteria for inclusion body myositis and myopathologic criteria for macrophagic myofasciitis. In the three cases macrophagic myofasciitis complicated the evolution of a known and painless inclusion body myositis and was diagnosed in a repeated deltoid biopsy because of the appearance of myalgia during the course of inclusion body myositis in all cases. The unexpected appearance of myalgia during the course of painless inclusion body myositis must arouse the suspicion of an association of another inflammatory muscle disease, macrophagic myofasciitis. PMID- 11404117 TI - The frequency of 17p11.2 duplication and Connexin 32 mutations in 282 Charcot Marie-Tooth families in relation to the mode of inheritance and motor nerve conduction velocity. AB - The 17p11.2 duplication and Connexin 32 (Cx32) mutations are the most frequent gene mutations responsible for Charcot-Marie-Tooth diseases. We classified 282 Charcot-Marie-Tooth families according to the median motor nerve conduction velocity of the index patient and the mode of inheritance, and screened them for 17p11.2 duplication and Cx32 mutations. Forty-seven percent of the Charcot-Marie Tooth families had median motor nerve conduction velocity under 30 m/s (group 1), 15% between 30 and 40 m/s (group 2), and 28% over 40 m/s (group 3). Spinal Charcot-Marie-Tooth (group 4) was observed in 7% of the families. Modes of inheritance were not similarly represented among the different groups. The 17p11.2 duplication was detected in index patients of group 1 only, and accounted for 83% of the familial cases and 36% of the isolated cases. In contrast, 21 Cx32 mutations were detected to variable degrees in groups 1-3, but were most numerous by far in dominant families of group 2 (44%). This systematic approach was taken to estimate the frequency of 17p11.2 duplication and Cx32 mutations in the different Charcot-Marie-Tooth subgroups, in order to propose a practical strategy for molecular analysis. PMID- 11404118 TI - Depressed myocardial fatty acid metabolism in patients with muscular dystrophy. AB - Myocardial involvement is frequently associated with various types of muscular dystrophy and Thallium-201 scintigraphy can show regional myocardial perfusion abnormalities in patients with muscular dystrophy. Myocardial fatty acid metabolism can now be imaged using a radioiodinated branched fatty acid (123I BMIPP). The present study evaluates myocardial fatty acid metabolism in muscular dystrophy. Twenty-eight patients underwent 123I-BMIPP(BMIPP) and Thallium dual single photon emission tomography. Regional uptake of both tracers was visually analyzed. We also assessed electrocardiography and echocardiography. The results showed that the BMIPP uptake compared to Thallium was smaller in 57% of all patients. BMIPP SPECT images revealed abnormalities in four of eight patients with a normal electrocardiogram. Abnormal BMIPP uptake with normal regional wall motion was evident in nine patients. The size of the region with defective BMIPP uptake was larger than that of asynergic areas detected by echocardiography in 11 patients. In conclusion, muscular dystrophy involves depressed myocardial fatty acid metabolism in larger extent of region than that in perfusion or mechanical abnormality. PMID- 11404119 TI - A mitochondrial encephalo-myo-neuropathy with a nucleotide position 3271 (T-C) point mutation in the mitochondrial DNA. AB - We report three members of a family, who exhibited a phenotype similar to 'myoclonus epilepsy with ragged-red fibers' but had a genotype usually associated with 'mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes'. The patients, a 48-year-old female, and her two sons, aged 21 and 19 respectively, presented with photo-reactive syncopal episodes, disturbances of gait and writing, dysarthria and finger tremor since the 3rd and 2nd decade of life, respectively, that were accompanied also by numbness and weakness of the extremities. Subsequently, cerebellar ataxia and myoclonus were also noted. Electromyography revealed both myogenic and neurogenic muscular changes, and nerve conduction studies demonstrated a sensory-motor neuropathy. Biopsy showed ragged-red fibers with strongly stained SDH-positive vessels in skeletal muscles, and a marked loss of myelinated fibers of the sural nerves. Mitochondrial (mt) DNA analyses of peripheral blood, muscles and nerves revealed that all members had a heteroplasmic np3271 (T-C) point mutation in the mitochondrial tRNA-Leu gene (UUR). This family is unique, in that all patients presented with a myoclonus epilepsy with ragged-red fibers-like phenotype and had a distinctive peripheral neuropathy, while the detected mtDNA 327l (T-C) mutation has been reported to date only in rare cases of mitochondrial encephalomyopathy with lactic acidosis and stroke-like episodes PMID- 11404120 TI - A new mtDNA mutation in the tRNA(Leu(UUR)) gene associated with ocular myopathy. AB - We studied a patient with ptosis, ophthalmoparesis, and exercise intolerance who showed in her muscle biopsy ragged-red fibers and combined defects of the complexes I and IV of the mitochondrial respiratory chain. Molecular analysis revealed a T3273C transition in the mitochondrial DNA tRNA(Leu(UUR)) gene. The mutation was heteroplasmic and very abundant in muscle from the proposita, less abundant in her other tissues studied, and still less abundant in blood from her maternal relatives. Single muscle fiber analysis showed significantly higher levels of mutant genomes in ragged-red fibers than in normal fibers. The T3273C mutation affects a strictly conserved base pair in the anticodon stem and was not found in controls, thus satisfying the accepted criteria for pathogenicity. PMID- 11404121 TI - A new mutation in the mitochondrial tRNA(Ala) gene in a patient with ophthalmoplegia and dysphagia. AB - We describe a new mutation in the tRNA(Ala) gene, a T-->C transition at nucleotide position 5628, in a 62-year-old woman with late onset chronic progressive external ophthalmoplegia, dysphagia and mild proximal myopathy. The mutation is heteroplasmic and disrupts a highly conserved A-U base pair within the anticodon stem of the tRNA(Ala). Cytochrome c oxidase-negative fibers harbor a significantly higher level of mutated mtDNA than cytochrome c oxidase-positive fibers. This is the first mutation in the tRNA(Ala) gene which satisfies accepted criteria for pathogenicity. PMID- 11404122 TI - Proximal myotonic myopathy and proximal myotonic dystrophy: two different entities? The phenotypic variability of proximal myotonic syndromes. AB - Multisystemic myotonic myopathies are characterised by a variable pattern of symptoms and signs and a variable degree of disease severity. Proximal myotonic dystrophy has been described as an entity distinct from proximal myotonic myopathy because of severe proximal muscle weakness and dystrophic changes on magnetic reasonace imaging and on muscle histopathology. We describe two siblings, one of them presenting with a proximal myotonic myopathy phenotype, the other with a proximal myotonic dystrophy-like phenotype. The variability of disease expression in these two siblings suggests that a proximal myotonic dystrophy-like variant may occur in proximal myotonic myopathy. PMID- 11404123 TI - Neonatal arthrogryposis and absent limb muscles: a muscle developmental gene defect? AB - We describe a child who presented at birth with arthrogryposis. Following a muscle biopsy a diagnosis of congenital muscular dystrophy was made and a skin biopsy 12 years later confirmed the presence of merosin. Her clinical picture was unusual, however, for merosin-positive congenital muscular dystrophy. She had extreme wasting and weakness of her arms and legs. In contrast, she had good neck and trunk control, and no facial or respiratory muscle weakness. We have used magnetic resonance imaging to examine the pattern of muscle involvement in this case. No recognizable muscle could be identified in the limbs. In contrast, the axial muscles were preserved. This striking pattern of virtual absence of muscles in the limbs with sparing of the axial muscle suggests that a gene responsible for the migration and/or proliferation of limb muscle precursor cells may be involved in the disease process. It is recognized that merosin-positive congenital muscular dystrophy is a heterogeneous disease. Magnetic resonance imaging is a useful tool for examining in detail the pattern of muscle involvement and identifying individual phenotypes. Understanding more about which muscles are affected in children with congenital myopathies may provide information on the underlying pathological process and help in the search for candidate proteins and genes. PMID- 11404124 TI - Pseudo-metabolic presentation in a Duchenne muscular dystrophy symptomatic carrier with 'de novo' duplication of dystrophin gene. AB - We report a 6-year-old female patient presenting with a sudden and severe single episode of rhabdomyolysis in which screening for a metabolic disorder was negative. Four months after the episode a muscle biopsy was performed and showed a mild pattern of necrosis/regeneration. Upon immunofluorescence, a mosaic pattern of dystrophin deficiency was found, and in the dystrophin deficient muscle fibres, the four proteins of the sarcoglycan complex were also lacking. Genetic analysis showed a duplication of exons 3 to 17 on one X-chromosome of the proband, but not on the mother's X-chromosome. A clearly skewed X-inactivation (85% of the defective X being active) was found and is consistent with the patient being symptomatic. To our knowledge, a spontaneous rhabdomyolysis in a female Duchenne muscular dystrophy carrier has never been reported. PMID- 11404125 TI - Endovascular treatment of acute bleeding complications in traumatic aortic rupture and aortobronchial fistula. AB - OBJECTIVE: Herein we report our experience in placement of endovascular stentgrafts in the descending aorta in patients with acute bleeding complications due to traumatic rupture or aortobronchial fistula. METHODS: Six patients (one woman, five men, mean age 47+/-19 years) were treated from September 1995 to February 2000 by implantation of endovascular stentgrafts in the descending aorta. Indications included traumatic ruptures of the aortic isthmus (n=3) and aortobronchial fistulas (n=3). All procedures were performed under general anaesthesia. The implants were introduced under fluoroscopic guidance via the aorta (n=1), the iliac (n=4) or femoral (n=2) artery, respectively. RESULTS: All aortobronchial fistulas and ruptures were sealed up successfully. There was no perioperative morbidity and no procedure-related morbidity except one patient who received aortofemoral reconstruction because of iliac occlusive disease. All patients are alive and well after a mean follow-up of 31 months (range 6-60). Two patients had recurrent hemoptysis, in one case, the patient received a second implant (distal extension), the other patient was managed conservatively. CONCLUSION: Endovascular treatment by a stentgraft is a safe and reliable procedure in the management of acute bleeding complications in patients with aortic rupture or aortobronchial fistulas. PMID- 11404126 TI - Apoptotic cell death in the hippocampus due to prolonged hypothermic circulatory arrest: comparison of cyclosporine A and cycloheximide on neuron survival. AB - OBJECTIVE: To determine whether cyclosporine A (CsA) or cycloheximide (CHX) can reduce neuronal apoptosis in the hippocampus in a chronic animal model of hypothermic circulatory arrest (HCA). METHODS: Twenty-eight pigs (28-33 kg) underwent 90 min of HCA at 20 degrees C. In a blinded study, animals were randomized to placebo (n=12), 5 mg/kg CsA (n=8), or 1 mg/kg CHX (n=8). After elective sacrifice 7 days postoperatively, brains were perfusion-fixed and the left hippocampus was examined for evidence of neuronal cell death. An in situ double-labeling method was used on cryosections to unequivocally identify apoptotic nuclei by the simultaneous visualization of DNA fragmentation and apoptotic chromatin condensation. Sections were also examined by immunocytochemistry for upregulation of the pro-apoptotic proteins Bax, activated caspase 3, and glyceraldehyde-3-phosphate dehydrogenase. RESULTS: Apoptotic nuclear degradation was clearly present in the CA1, CA2 and CA3 subregions of the hippocampus after HCA. However, there was also morphological evidence for an accompanying necrotic-like cell death. There was no significant difference between the number of apoptotic nuclei observed in CSA-treated animals, mean value 4.4+/-1.63 SEM or CHX-treated animals, mean value 4.0+/-1.92 SEM, and age matched control HCA pigs, mean value 4.85+/-1.69 SEM, (P>0.10). CONCLUSIONS: The data clearly demonstrate apoptotic cell death in pigs after HCA by simultaneously demonstrating in situ end labeling (TUNEL reaction) and apoptotic chromatin condensation using a nucleic acid-binding dye. Since CsA shows promising neuroprotective effects in behavioral studies, and since the peak of HCA-induced apoptosis occurs earlier than 7 days, further studies will be required to determine whether CsA can improve neuronal survival in the first few days after HCA. CHX was not effective in reducing apoptosis in this model. PMID- 11404127 TI - Cyclosporine A as a potential neuroprotective agent: a study of prolonged hypothermic circulatory arrest in a chronic porcine model. AB - OBJECTIVE: To assess whether Cyclosporine A (CsA) or cycloheximide (CHX) can reduce ischemia-induced neurological damage by blocking apoptotic pathways, we assessed their effects on cerebral recovery in a chronic animal model of hypothermic circulatory arrest (HCA). METHODS: Twenty-eight pigs (28-33 kg) underwent 90 min of HCA at 20 degrees C. In this blinded study, animals were randomized to placebo (n=12), 5 mg/kg CsA (n=8), given intravenously before and subcutaneously for 7 days after HCA, or a single dose of 1 mg/kg CHX (n=8), given after weaning from cardiopulmonary bypass. Hemodynamics, intracranial pressure (ICP) and neurophysiological data (EEG, SSEP) were assessed for 3 h after HCA; early behavioral recovery was scored, and neurological/behavioral evaluation (9=normal) was carried out daily until elective sacrifice on postoperative day (POD) 7. Brains were selectively perfused and evaluated histopathologically for apoptosis. RESULTS: Basic hemodynamic data revealed no differences between CsA or CHX and control groups. ICP was significantly lower throughout rewarming (P=0.009) and reperfusion (P=0.05) in the CsA group. EEG recovery 3 h after HCA was observed in four of eight CsA animals but in only 1 of 12 controls (P=0.11) and one of eight CHX animals; cortical SSEP recovery also seemed faster in CsA animals, but failed to reach significance. Some early recovery scores were significantly better in the CsA group, and daily behavioral scores were consistently and significantly higher in the CsA-treated animals from POD1 through POD4. CONCLUSIONS: The data indicate that treatment with Cyclosporine A but not cycloheximide has a positive effect on cerebral recovery following HCA. Whether CsA results in inhibition of neuronal apoptosis, and/or inhibits release of cytokines and thereby reduces postischemic cerebral edema remains to be elucidated. The neuroprotective effect of CsA, if confirmed in further studies, would make its clinical application conceivable. PMID- 11404128 TI - Antegrade selective cerebral perfusion during surgery of the thoracic aorta: risk analysis. AB - OBJECTIVE: To determine independent predictors of neurologic outcome and hospital mortality after surgery of the thoracic aorta using moderate hypothermic circulatory arrest and antegrade selective cerebral perfusion. METHODS: Between November 1996 and June 2000, 96 consecutive patients (69 men, 27 women; mean age 63+/-10 years) underwent operations on the thoracic aorta with the aid of moderate hypothermic circulatory arrest and antegrade selective cerebral perfusion. Sixty-four patients were operated on electively (66.7%), 32 emergently (33.3%). Indications for surgery were: type A acute dissection in 30 patients (31.3%), chronic aneurysm in 66 (68.8%). Seventeen patients (17.7%) had undergone previous aortic/cardiac surgical procedures. The mean selective cerebral perfusion time was 52.2+/-31.9 min (range, 18-220 min). Preoperative, intraoperative, and postoperative factors were analyzed by univariate and multivariate analysis to identify predictors of hospital mortality and neurologic outcome. RESULTS: There were no operative deaths; the hospital mortality rate was 11.5% (11/96). Stepwise logistic regression revealed preoperative renal dysfunction (P=0.021), type A acute dissection (P=0.053), coronary artery bypass grafting (P=0.058), post-operative pulmonary complications (P=0.000) and repeat thoracotomy for bleeding (P=0.027) as independent predictors of hospital mortality. One patient sustained a permanent neurologic deficit (1%). Transient neurologic deficit occurred in eight patients (8.3%). Coronary artery bypass grafting (P=0.013), and postoperative cardiac complications (P=0.049) were statistically associated with an increased risk of any (transient and permanent) neurologic dysfunction on univariate analysis. Stepwise logistic regression indicated coronary artery bypass grafting as independent factor for any neurologic dysfunction. CONCLUSION: This study confirmed that selective cerebral perfusion is an effective method of cerebral protection allowing complex thoracic aorta operations to be performed with low risk of hospital mortality and adverse neurologic outcome. We didn't find that the duration of selective cerebral perfusion time influence hospital mortality and any neurologic deficit. PMID- 11404129 TI - Pulmonary thromboendarterectomy--risk factors for early survival and hemodynamic improvement. AB - OBJECTIVE: Pulmonary thromboendarterectomy (PTE) for chronic thromboembolic pulmonary hypertension is a challenging procedure with a considerable mortality. The aim of this investigation was to identify risk factors influencing mortality and operative results. METHODS: Between October 1995 and August 2000, 69 patients (age 54 years; 34 women; mean New York Heart Association (NYHA) stage 3.4) underwent PTE. The preoperative pulmonary vascular resistance (PVR) was 988+/-554 dynes x s x cm(-5), mean pulmonary artery pressure 50+/-12 mmHg, right atrial pressure (RAP) 11.5+/-4 mmHg. Hospital mortality was 10.1% (n=7/69). Mean postoperative PVR on the 2nd day was 324+/-188 dynes x s x cm(-5). Pulmonary angiography was reviewed for number of involved segments (mean 9.3+/-2) and bronchial arteries diameter (BAD; mean 4.6+/-1.6 mm). A univariate and multivariate analysis was performed to determine preoperative risk factors for hospital death and inadequate hemodynamic improvement. RESULTS: By univariate analysis, mortality was influenced by age (P=0.04), right atrial pressure (P=0.009), NYHA (P=0.02) and the number of angiographically involved segments (P=0.02). Sex, left ventricular function, presence of coronary artery disease and bronchial artery diameter did not show correlation with mortality. Inadequate hemodynamic improvement in a dichotomized analysis (PVR > or =500 dynes x s x cm( 5), n=11, and PVR < 500 dynes x s x cm(-5), n=58), assessed by univariate analysis, was significantly influenced by age (P=0.02), preoperative PVR (P=0.01), NYHA (P=0.002), RAP (P=0.02) and female sex (P=0.02). Multivariate analysis identified age (P=0.1), RAP (P=0.002) and female sex (P=0.007) as risk factors for inferior hemodynamic improvement. CONCLUSIONS: Preoperative parameters can be utilized to assess postoperative mortality and hemodynamic improvement after pulmonary thromboendarterectomy. Patient age and clinical deterioration of pulmonary hypertension are considerable preoperative factors influencing hospital mortality. Inadequate postoperative hemodynamic improvement is affected by severity of disease and female sex. PMID- 11404130 TI - Tracheal surgery in children: an 18-year review of four techniques. AB - OBJECTIVE: Review the short- and long-term outcomes of a single institution experience in infants with congenital tracheal stenosis, comparing four different operative techniques used from 1982 through 2000. METHODS: Hospital and clinic records of 50 infants and children who had surgical repair of congenital tracheal stenosis secondary to complete tracheal rings were reviewed. Age at surgery ranged from 7 days to 72 months (median, 5 months, mean 7.8+/-12 months). Techniques included pericardial patch tracheoplasty (n=28), tracheal autograft (n=12), tracheal resection (n=8), and slide tracheoplasty (n=2). All procedures were done through a median sternotomy with cardiopulmonary bypass. Seventeen patients had a pulmonary artery sling (35%), and 11 had an intracardiac anomaly (22%). RESULTS: There were three early deaths (6% early mortality), two after pericardial tracheoplasty and one after autograft. There were six late deaths (12% late mortality), five after pericardial tracheoplasty and one after slide tracheoplasty. Length of stay (median) was 60 days (pericardial tracheoplasty), 28 days (autograft), 14 days (resection), and 18 days (slide). Reoperation and/or stent placement was required in seven patients (25%) after pericardial tracheoplasty, in two patients (17%) after autograft, in no patients after resection, and in one patient (50%) after slide tracheoplasty. CONCLUSIONS: Our current procedures of choice for infants with congenital tracheal stenosis are resection with end-to-end anastomosis for short-segment stenoses (up to eight rings) and the autograft technique for long-segment stenoses. Associated pulmonary artery sling and intracardiac anomalies should be repaired simultaneously. PMID- 11404131 TI - Effect of fenestration on the sub-diaphragmatic venous hemodynamics in the total cavopulmonary connection. AB - OBJECTIVE: To understand differences in the sub-diaphragmatic venous physiology between patients with fenestrated and non-fenestrated total-cavopulmonary connections (TCPC). METHODS: We studied the effects of respiration, retrograde flow, and gravity on the sub-diaphragmatic venous flows in 20 normal healthy volunteers (control), 25 Fontan patients with non-fenestrated TCPC, and 21 with fenestrated TCPC. Subhepatic inferior vena cava (IVC), hepatic vein (HV), and portal vein (PV) flow rates were measured with Doppler ultrasonography during inspiration and expiration in both supine and upright positions. The supine inspiratory-to-expiratory flow rate ratio was calculated to reflect the effect of respiration, the supine-to-upright flow rate ratio was calculated to assess the effect of gravity, and the magnitude of retrograde flow was evaluated with respect to total antegrade flow. Mean IVC, HV, and wedged hepatic venous (WHV) pressures were measured during cardiac catheterization in four TCPC patients before and after fenestration closure. The transhepatic venous pressure gradient (TVPG) was calculated as the difference between the HV and WHV pressure. RESULTS: Compared with control, HV flow in TCPC was heavily dependent on respiration; this inspiratory capacity was greater in fenestrated than non-fenestrated subjects (inspiratory-to-expiratory flow ratio 1.7, 4.4, and 3.0, respectively P<0.001). Normal retrograde HV flow was diminished in TCPC patients, furthermore, fenestrated subjects had less flow reversal than non-fenestrated (retrograde as percent of antegrade flow 43, 19, and 30%, respectively P<0.001). Gravity decreased IVC and HV flows more in TCPC subjects than control, but this effect was not different between the two TCPC groups. Closure of the fenestration resulted in higher IVC and HV pressures (pre-closure versus post-closure pressures [mmHg]: 11.2 +/- 4.0 vs. 12.3 +/- 3.9, and 11.5 +/- 3.8 vs. 12.4 +/- 3.8, respectively P< or =0.001). The normal TVPG was reduced in fenestrated TCPC, and worsened after fenestration closure (0.9 +/- 0.3 and 0.7 +/- 0.4, respectively P < 0.04). CONCLUSIONS: Fenestration of the inferior venous connection has important influences on sub-diaphragmatic venous return in TCPC patients. Although fenestration lowers venous pressures and partially restores TVPG, its beneficial effects on flow in TCPC patients are mediated primarily by an increase in inspiration-derived forward HV flow and reduced flow reversal. These observations suggest fenestration results in a more efficient and less congested splanchnic circulation in TCPC patients, and may have important implications in the early and late management of Fontan patients. PMID- 11404132 TI - The Konno aortoventriculoplasty for repeat aortic valve replacement. AB - OBJECTIVE: To evaluate the outcome of aortic root augmentation by the Konno aortoventriculoplasty technique as part of reoperative aortic valve replacement. METHODS: Since 1983, 15 patients, 12 males and three females, had repeat aortic valve replacement (AVR) with concomitant Konno aortoventriculoplasty. Age ranged from 1.2 to 18 years (mean 12.5 years). The underlying anatomic diagnoses were valve and subvalvar aortic stenosis in 11, truncal valve insufficiency in one, endocarditis in one, Shone's complex in one and severe aortic insufficiency associated with a ventricular septal defect in one patient. All patients had had previous AVR. The causes for reoperation were prosthetic valve stenosis due to growth in ten and paravalvular leak in one, homograft failure in two, xenograft failure in one, and left ventricular outflow tract obstruction (LVOTO) after mitral valve replacement in one patient. The mean size of explanted prostheses was 19.2 mm (13-23 mm) while the mean size of the implanted prostheses was 24.3 mm (19-27 mm) (P<0.01). Previous aortic root enlargement had been performed in 11 patients in conjunction with AVR. The Manougian technique was used previously in two, Konno aortoventriculoplasty in eight, and both techniques in one patient. The newly implanted aortic valves were a homograft in one patient and mechanical prostheses in 14 patients. RESULTS: There was one operative death (1 of 15 or 6.6%) in a 17.5 year old patient with previous AVR and posterior root enlargement, due to low cardiac output state. Follow-up ranged from 6 months to 17 years (mean 7.2 years). The only late death occurred in an 11.6-year-old patient due to prosthetic valve endocarditis. Two patients had complete heart block and had permanent pacemaker insertion (2 of 15 or 13.3%). One patient had pulmonary valve replacement because of combined stenosis and insufficiency 5 years after operation. All 13-surviving patients are asymptomatic at latest follow up. CONCLUSION: Konno aortoventriculoplasty with repeat AVR may be safely performed. Excellent results may be achieved despite previous aortic root enlargement. It is a good surgical option for complex LVOTO and may even reduce reoperation in children by allowing placement of a larger prosthesis. PMID- 11404133 TI - Left ventricular architecture after valve replacement due to critical aortic stenosis: an approach to dis-/qualify the myth of valve prosthesis-patient mismatch? AB - OBJECTIVES: Left ventricular hypertrophy in patients with critical aortic stenosis (AS) is an adaptive process that compensates for high intracavitary pressure and reduces systolic wall stress followed by an increase in myocardial masses. In the present prospective clinical trial, we investigated long-term compensatory changes in left ventricular geometry and function after aortic valve replacement using mechanical bileaflet prostheses with the main emphasis on the small-sized aortic annulus and valve prosthesis-patient mismatch. METHODS: A total of 58 patients with critical AS were assigned to the following groups according to the predictive value of prosthetic valve area index (VAI): group EXMIS: 29 patients (VAI < or =0.99), expected mismatch; group NOMIS: 29 patients (VAI < or =0.99), no mismatch. At controls T(0) (before operation/operation (OP), T(1) and T(2) (4 and 20 months after OP) the left ventricular geometry was recorded by means of Imatron electron beam tomography and the transprosthetic velocities were measured by echocardiography. RESULTS: Statistical analysis showed a consistent reduction in the absolute (P=0.04) and indexed (P=0.04) left ventricular myocardial mass for both cohorts; furthermore, there was a significant difference between EXMIS and NOMIS patients concerning the factors, time and mass reduction (P=0.005), because of distinct baselines. A logistic regression report revealed preoperative cardiac output, absolute left ventricular myocardial mass, perfusion, body surface area and the native valve orifice area as predicting coefficients and factors for a minimum mass reduction of 25%. We explain a mathematical formula that turned out to be the most sensitive for correctly classified factors. CONCLUSIONS: The left ventricular geometry and transprosthetic velocities resulted in the same postoperative recovery for both EXMIS and NOMIS patients. The presented data showed that valve prosthesis-patient mismatch had no influence in several stepwise logistic regression models. We conclude that modern mechanical bileaflet prostheses allow both acceptable hemodynamics and recovery of left ventricular hypertrophy, even in small aortic annuli. PMID- 11404134 TI - Successful management of Brucella mellitensis endocarditis with combined medical and surgical approach. AB - OBJECTIVES: Brucella endocarditis is an underdiagnosed complication of human brucellosis, associated with high morbidity and mortality. We report the successful management of a number of cases of Brucella mellitensis endocarditis. PATIENTS AND METHODS: Seven consecutive cases of Brucella mellitensis endocarditis were treated over the last 20 years, based on high suspicion of the disease at first place. The early suspicion of Brucella endocarditis relied on medical history and a standard tube agglutination titer > or =20. Blood and/or cardiac tissue cultures were positive in all patients, but available late following surgery. All patients were successfully treated with a combination of aggressive medical and early surgical therapy. All affected valves were replaced within 1 week from admission (five aortic and three mitrals). Medical treatment included co-trimoxazole, tetracyclines and streptomycin, before surgery, followed by co-trimoxazole and tetracyclines for a median of 12 months (range: 3-15 months) after surgery until the titers returned to a level < or =1:160. RESULTS: There were neither operative deaths nor recurrence of infection. One patient died two years after the operation due to massive cerebrovascular accident. Ten-year survival was 85.7+/-13.2%. CONCLUSION: Although Brucella mellitensis endocarditis is a rare entity, its optimum management should be a combination of aggressive medical treatment and early surgical intervention, based on high degree of suspicion in areas with high incidence of the disease. PMID- 11404135 TI - Deep sternal wound infection: the role of early debridement surgery. AB - OBJECTIVE: This retrospective chart review study aimed to evaluate whether a more aggressive staged approach can reduce morbidity and mortality following post cardiotomy deep sternal wound infection. METHODS: Between 1979 and 2000, 14620 patients underwent open heart surgery: mediastinitis developed in 124 patients (0.85%). Patients were divided in two groups: in 62 patients (Group A) (1979 1994) an initial attempt of conservative antibiotic therapy was the rule followed by surgical approach in case of failure; in 62 patients (Group B) (1995-2000) the treatment was staged in three phases: (1) wound debridement, removal of wires and sutures, closed irrigation for 10 days; (2) in case of failure open dressing with sugar and hyperbaric therapy (11 patients, 17%); (3) delayed healing and negative wound cultures mandated plastic reconstruction (three patients, 4%). Categorical values were compared using the Chi-square test, continuous data were compared by unpaired t-test. RESULTS: Incidence of mediastinitis was higher in Group B (62 out of 5535; 1.3%) than in Group A (62 out of 9085; 0.7%) (P=0.007). Mean interval between diagnosis and treatment was shorter in Group B (18+/-6 days) than in group A (38+/-7 days) (P=0.001). Hospital mortality was higher in Group A (19/62; 31%) than in Group B (1 out of 62; 1.6%) (P<0.001). Hospital stay was shorter in Group B (30.5+/-3 days) than in group A (44+/-9 days) (P=0.001). In Group B complete healing was observed in all the 61 survivors: 47 cases (76%) after Stage 1; 11 (18%) after Stage 2; three (4.8%) after Stage 3. CONCLUSIONS: Although partially biased by the fact that the two compared groups draw back to different decades, this study showed that an aggressive therapeutic protocol can significantly reduce morbidity and mortality of deep sternal wound infection. PMID- 11404136 TI - The relationship between predicted and actual cardiac surgical mortality: impact of risk grouping and individual surgeons. AB - OBJECTIVE: To study the relationship between predicted and actual mortality in a cardiac surgical practice and to determine whether there is a consistent relationship across risk groups and surgeons. METHODS: Risk information (Parsonnet score) was prospectively collected for 6213 consecutive adult patients undergoing cardiac surgery at one institution. The relationship between predicted mortality and actual mortality was analysed by risk group for all patients and for individual surgeons' practices. RESULTS: Predicted mortality was 10.2%. Actual mortality was 4.2%, giving a mortality ratio of 41% of predicted. This ratio was not consistent across the five major risk groups, ranging from 32% in moderate risk to 67% in very low risk patients. When analysed by individual surgical practices, the results were even more disparate, with a mortality index range between 0% for one surgeon's low risk patients to 150% for another surgeon's very low risk patients. CONCLUSION: The relationship between predicted and actual mortality at one institution may vary across the risk spectrum and between surgeons. This should be taken into account in preoperative risk assessment and informed patient consent. Individual surgeons may have strengths and weaknesses which are related to preoperative risk stratification. PMID- 11404137 TI - Nodal vessels disease as a risk factor for atrial fibrillation after coronary artery bypass graft surgery. AB - OBJECTIVE: Atrial fibrillation (AF) is common after coronary artery bypass graft (CABG) surgery. Atrial ischaemia due to diseased atrioventricular (AV) and sinoatrial (SA) arteries has been proposed as a cause of AF post-CABG. We examined if the presence of diseased nodal arteries was a significant predictor of the development of AF post-CABG. METHODS: 100 consecutive cases (AF post-CABG) were compared to 100 consecutive controls (No AF post-CABG) with respect to pre operative angiographic evidence of diseased nodal arteries. Cases and controls identified from the Society of Thoracic Surgeons database underwent detailed chart reviews to obtain data on potential risk factors. Patients were excluded if they were undergoing anything but a routine CABG procedure, were older than 65 years, or had previous AF. All angiograms were reviewed by a single radiologist blinded to outcome. The effect of grafting diseased nodal arteries on the development of AF post-CABG was also measured. A multiple logistic regression model was utilized to measure the effect of disease in each artery on the development of AF post-CABG. RESULTS: Cases and controls were comparable regarding potential risk factors, with the exception that the AF group was older than the non-AF group. Significant AV artery disease was detected in 78 cases compared to 74 controls (adjusted odds ratio (OR) OR=1.04, CI, 0.51-2.12, P=0.82). Significant SA artery disease was detected in 34 cases compared to 21 controls (adjusted OR=2.093, CI: 1.06-4.09, P=0.03). Six of ten patients having revascularization of their SA nodal artery developed AF versus 28 of 45 of those who did not (OR=0.91, CI: 0.18-5.06, P=0.58). Forty-eight of 87 patients having revasacularization of their AV nodal artery developed AF versus 30 of 65 of those who did not (OR=1.44, CI: 0.72-2.88, P=0.27). CONCLUSION: The presence of a diseased SA artery is significantly associated with AF post-CABG. Such association may be used to identify a subset of patients who might be targeted with prophylaxis. PMID- 11404138 TI - Respiratory dysfunction after coronary artery bypass grafting employing bilateral internal mammary arteries: the influence of intact pleura. AB - OBJECTIVE: To evaluate the role of intact pleurae regarding the postoperative respiratory functional status in patients undergoing coronary revascularization employing both internal mammary arteries (IMAs), according to the pedunculated or skeletonized technique (SKT) with opened or intact pleurae. MATERIALS AND METHODS: Using both IMAs, 299 patients underwent elective coronary revascularization. They were randomized and divided into group I (n=82, undergoing IMA harvesting according to the SKT without opening the pleurae); group II (n=186, undergoing IMA harvesting according the pedunculated technique with open pleurae); and group III (n=31, undergoing IMA harvesting according the SKT with incidentally opened pleurae). There were no differences regarding the preoperative patient characteristics and the anaesthetic and surgical management. RESULTS: There were two deaths in group I versus seven in group II and one in group III (P=ns). The number of total arterial myocardial revascularization and arterial composite grafts was significantly higher in groups I and III than in group II, (P<0.001 and P<0.005, respectively). The incidence of postoperative complications was similar between groups. Blood loss of >1000 ml was significantly higher in group II than group I (P<0.028); but the incidence of re thoracotomy and blood transfusion was similar between groups. The mechanical ventilation time was significantly higher in groups II and III versus group I (P<0.018 and P<0.02, respectively). The incidence of prolonged ventilation (>24 h), pleural effusion, thoracocentesis and atelectasis, resulted in being significantly higher in group II than group I. The incidence of thoracocentesis was significantly higher in group III than group I. The pain score and analgesic requirements at 1-12 h after awakening were significantly higher in groups II and III versus group I, becoming similar after the chest tubes were removed. PaO(2) was significantly higher, and PaCO(2) and FiO(2) were significantly lower in group I than groups II and III at 1 and 4 h before extubation and at 1 and 4 h after extubation. PaO(2) and PaCO(2) became similar between groups at the 5th postoperative day. CONCLUSIONS: According to our results, we may conclude that pleural integrity has beneficial effects on the respiratory functional status after coronary revascularization using both IMAs. A meticulous and more careful IMA harvesting approach significantly reduces the postoperative morbidity regarding the pulmonary functional status, and as a consequence, reduces the hospital costs. PMID- 11404139 TI - Coronary endothelial damage during off-pump CABG related to coronary-clamping and gas insufflation. AB - OBJECTIVE: Although off-pump coronary artery bypass grafting (CABG) has been recognized less invasive than conventional CABG on cardiopulmonary bypass, off pump CABG may be partly invasive especially to the coronary endothelium. The present study was designed to evaluate the adverse effects of coronary snaring with looped sutures and gas insufflation on the coronary endothelium. The protective efficacies on the coronary endothelium of coronary snaring with elastic sutures or humidified gas insufflation with/without heparin and dipyridamole-added were also tested. METHODS: Thirty-six mongrel dogs were used. After systemic heparinization (150 U/kg), a 5 mm longitudinal coronary incision was made with looped non-elastic monofilament sutures or elastic sutures applied proximally and distally. The incised coronary artery was exposed to non humidified carbon dioxide, humidified carbon dioxide with lactated Ringer solution, or humidified carbon dioxide with heparin and dipyridamole-added lactated Ringer solution for 10 or 20 min in each group. After gas insufflation, the incised coronary artery was repaired, then, the coronary was reperfused. Perfusion-fixation was done for observation of the coronary endothelium by scanning electron microscopy. The adverse effect on the endothelium was graded as follows: grade 1, appeared normal; grade 2, few blood cells deposited; grade 3, many blood cells deposited; grade 4, few endothelial cells delaminated with blood cells deposited; grade 5, many endothelial cells delaminated with blood cells deposited. RESULTS: Non-elastic looping caused much more endothelial tears than elastic looping (P<0.00001). Non-humidified gas blowing for 20 min caused more endothelial cell damage than humidified gas blowing (P=0.00005). Non-humidified gas blowing for 10 min caused less damage than for 20 min (P=0.00326), but still caused more damage than humidified gas blowing (P=0.00253). Heparin and dipyridamole-added humidification reduced coronary endothelial area mottled by the deposited cells when compared with simple humidification (P=0.00120). CONCLUSIONS: Coronary snaring resulted in coronary endothelial injury, which was ameliorated by using elastic sutures instead of non-elastic sutures. Non humidified gas insufflation made blood cells deposited and endothelial cells delaminated with time. Humidified gas insufflation attenuated these adverse effects. Heparin and dipyridamole-added humidification had potential advantage in terms of reducing deposited blood cells on the endothelium over simple humidification. PMID- 11404140 TI - Coronary flow reserve and nitric oxide synthases after cardiac transplantation in humans. AB - OBJECTIVE: Coronary endothelial dysfunction may precede morphological changes in both the epicardial conduit and microvascular resistance vessels in heart transplant recipients. Since the development of transplant atherosclerosis is the major limiting factor for long-term survival, the identification of early mediators of vasomotor dysfunction may be of therapeutic interest. We therefore investigated the potential relationship between the expression of nitric oxide synthases (NOS) and coronary endothelial function in human cardiac transplant recipients over time. METHODS: Forty-two human cardiac transplant recipients were studied at 1 and 12 months after heart transplantation (HTx). The microvascular coronary flow velocity reserve (CFVR) was tested for endothelium-dependent (acetylcholine) and -independent (adenosine) stimuli by intravascular Doppler flow-wire. Epicardial diameter changes were evaluated by quantitative coronary angiography. Endomyocardial inducible (iNOS) and endothelial constitutive nitric oxide synthase were determined by RT-PCR. Nitric oxide production (nitrite and nitrate (NOx)) and TNF-alpha were measured in plasma samples from the aorta and coronary sinus. RESULTS: CFVR was impaired in 26.1% (n=11) of patients at 1 month and in 31% (n=13) 12 months after HTx. iNOS-mRNA levels were significantly higher in patients with impaired endothelium-dependent CFVR. In addition, only in these patients were TNF-alpha levels higher and these correlated with plasma NOx levels at 1 and 12 months post-HTx (1 month: r=0.81, P=0.001; 12 months: r=0.62, P=0.04). CONCLUSIONS: Coronary microcirculatory dysfunction in response to acetylcholine is present in nearly 30% of patients during the first year following transplantation. These patients present with higher iNOS-mRNA expression and TNF-alpha plasma levels. Selective modulation of the TNF alpha/iNOS-pathway may be of therapeutic value to improve coronary endothelial dysfunction in cardiac transplant recipients. PMID- 11404141 TI - An evaluation of the intraoperative transit time measurements of coronary bypass flow. AB - OBJECTIVE: The intraoperative measurement of the coronary bypass flow enables the identification of technical errors while the sternum is still open. The transit time flow method is able to effectively measure the internal thoracic artery graft flow. The aim of the present study was to analyze the factors which affected the bypass flow rate. METHODS: We measured the blood flow of 291 in situ internal thoracic artery (ITA) and 190 saphenous vein (SV) grafts constructed in 171 patients undergoing coronary artery bypass grafting from December 1996 to March 2000 using this method during the surgery. All patients underwent postoperative coronary angiography before the patients were discharged. The blood flow rate of all bypass grafts constructed was assessed after the patients were weaned from cardiopulmonary bypass. RESULTS: The mean flow rate of all ITA grafts was 65.1+/-36.7 ml/min and that of all SV grafts was 56.4+/-29.9 ml/min. According to analyses using correlation tests, the graft flow was found to significantly correlate with the grafted perfusion areas and the diameter of the bypassed coronary arteries. However, no significant difference was observed between the flow rates of the ITA grafts with and without stenosis or string phenomenon, but significant (P<0.0001) correlation was observed between the occurrence of a string sign and the degree of proximal stenosis of the recipient coronary artery. Regarding SV grafts, the mean flow rate of occluded grafts (29.2+/-20.5 ml/min) was significantly (P<0.0001) less than non-occluded grafts (56.4+/-29.9 ml/min). CONCLUSIONS: The bypass flow was affected by such a large number of factors that only measuring the bypass flow rate could not sufficiently predict either stenosed or narrowed grafts. However, ITA grafts bypassed to the coronary arteries with less stenosis were shown to more easily become narrowed. PMID- 11404142 TI - Early angiographic control of perioperative ischemia after coronary artery bypass grafting. AB - OBJECTIVE: To assess the impact of immediate angiography in patients with defined clinical and laboratory criteria of perioperative myocardial infarction after coronary artery bypass operation. PATIENTS AND METHODS: Between January 1999 and December 1999 2052 patients underwent coronary artery bypass grafting in our institution. Out of this cohort 131 (6.4%) patients met the criteria of perioperative myocardial ischemia, which was defined as: (a) increase in the isoenzyme ratio of creatinine phosphokinase (CK/CK-MB] above 10%; (b) ischemic electrocardiographic episodes (defined as a new onset of elevated ST-segment change lasting at least 1 min and involving a shift from baseline of greater than or equal to 0.1 mV of ST-depression and a new association of a postoperative Q; (c) recurrent episodes of, or sustained ventricular tachyarrhythmia as well as ventricular fibrillation; (d) hemodynamic deterioration despite adequate inotropic support. RESULTS: Angiography was performed in 108 patients (5.3%, group A) whereas 23 patients (1.1%, group B) were immediately re-operated due to severely compromised hemodynamics. Angiographic results in group A showed regular grafts in 45 patients (2.2%); 63 patients (3.1%) had either an occlusion (n=41), incorrect anastomosis (n=29), graft stenosis (n=14), graft spasm (n=6), displaced graft (n=6), poor distal run-off (n=5) or incomplete revascularization (n=2). In group A 43 patients underwent a re-operation (34 patients) or an early angioplasty (nine patients). Due to poor coronary artery status no intervention was performed in the remaining 20 patients with angiographic findings. Operative findings in group B showed graft occlusion in ten patients (43.5%), incorrect anastomosis in five patients (21.7%), bleeding, stretched graft, venous graft spasm and displaced graft in one patient (4.3%) each, and no patho-morphological finding in 4 patients (17.4%). Thirty-day mortality rate was ten patients in group A (9.3%), all of them with angiographic findings, as opposed to nine patients (39.1%) in group B. CONCLUSION: ST-change and elevated CK/CK-MB enzyme ratio is highly indicative for possible graft failure and should be followed early angiographic control to assess the need for reintervention. PMID- 11404143 TI - Heart-type fatty acid binding protein (hFABP) in the diagnosis of myocardial damage in coronary artery bypass grafting. AB - OBJECTIVES: Heart-type fatty acid binding protein (hFABP) is an intracellular molecule engaged in the transport of fatty acids through myocardial cytoplasm and has been used as a rapid marker of myocardial infarction. However, its value in the evaluation of perioperative myocardial injury has not yet been assessed. METHODS: 32 consecutive patients undergoing coronary artery bypass grafting were included in a prospective, randomized study using standardized operative procedures and myocardial protection. Three patients with perioperative myocardial infarction were added. Serial blood samples were taken preoperatively, before ischemia, 5 and 60 min after declamping, 1 and 6 h postoperatively and on postoperative days 1, 2 and 10 and were tested for hFABP, creatine kinase isoenzyme MB (CKMB) and troponin I (TnI). RESULTS: Hospital mortality was zero. The kinetics of the biochemical parameters revealed a typical pattern for each marker. In routine patients, hFABP levels peaked as early as 1 h after declamping, whereas CKMB and TnI peaked only 1 h after arrival in the intensive care unit. Patients with perioperative infarction displayed peak levels some hours later in all marker proteins. Peak serum levels of hFABP correlated significantly with peak levels of CKMB (r=0.436, P=0.011) and TnI (r=0.548, P=0.001), indicating the degree of myocardial damage. CONCLUSIONS: hFABP is a rapid marker of perioperative myocardial damage and peaks earlier than CKMB or TnI. The kinetics of marker proteins in serial samples immediately after reperfusion is more suitable for the detection of perioperative myocardial infarction than a fixed cut-off level. PMID- 11404144 TI - Pyruvate/dichloroacetate supply during reperfusion accelerates recovery of cardiac energetics and improves mechanical function following cardioplegic arrest. AB - OBJECTIVES: Cardioplegic arrest during cardiac surgery induces severe abnormalities of the pyruvate metabolism, which may affect functional recovery of the heart. We aimed to evaluate the effect of pyruvate and dichloroacetate administration during reperfusion on recovery of mechanical function and energy metabolism in the heart subjected to prolonged cardioplegic arrest. METHODS: Four groups of rat hearts perfused in working mode were subjected to cardioplegic arrest (St. Thomas' No. 1), 4 h of ischaemia at 8 degrees C and reperfusion with either Krebs buffer alone (C) or with 2.8 mM pyruvate (P), with 1 mM dichloroacetate (D), or with a combination of both (PD). Mechanical function was recorded before cardioplegic arrest and at the end of experiments. In groups C and PD, additional experiments were performed using (31)P nuclear magnetic resonance spectroscopy in non-working Langendorff mode to evaluate cardiac high energy phosphate concentration changes throughout the experiment. RESULTS: Improved recovery of cardiac output (% of the preischaemic value+/-SEM, n=9-12) was observed in all three treated groups (65.7+/-4.3, 59.5+/-5.2 and 59.5+/-5.3% in PD, P and D, respectively) as compared with C (42.2+/-4.6%; P<0.05). Recovery of coronary flow was improved from 66.4+/-3.8 in C to 94.9+/-8.6% in PD (P<0.05). The phosphocreatine recovery rate in the first minutes of reperfusion was increased from 9.9+/-1.5 in C to 31.5+/-4.3 micromol/min per g dry wt in PD (P<0.001). No differences were observed in ATP or phosphocreatine concentrations at the end of experiment. CONCLUSIONS: The administration of pyruvate and dichloroacetate improves the recovery of mechanical function following hypothermic ischaemia. Accelerated restoration of the energy equilibrium in the initial phase of reperfusion may underlie the metabolic mechanism of this effect. PMID- 11404145 TI - L-Arginine given after ischaemic preconditioning can enhance cardioprotection in isolated rat hearts. AB - OBJECTIVE: Ischaemic or pharmacological preconditioning with L-arginine has been reported to be insufficient for optimal cardioprotection. The ability of nitric oxide (NO) to enhance ischaemic preconditioning was assessed, and the role of L arginine-induced ischaemic preconditioning in myocardial protection was determined. METHODS: Isolated rat hearts were prepared and divided into six groups: control hearts (control, n=6) were perfused without global ischaemia at 37 degrees C for 160 min; global ischaemia hearts (GI, n=6) were subjected to ischaemia for 20 min and reperfusion for 120 min; ischaemic preconditioned hearts (IP, n=6) received 2 min of zero-flow global ischaemia followed by 5 min reperfusion, before 20 min of global ischaemia; L-arginine hearts (ARG, n=6) received 1 mmol/l L-arginine for 5 min, before 20 min of global ischaemia; ischaemic preconditioning plus nitro-L-arginine methyl ester hearts (IP+L-NAME, n=6) received 2 min of ischaemic preconditioning and 5 min reperfusion with 3 mmol/l L-NAME in Krebs-Henseleit buffer, before 20 min of global ischaemia; and ischaemic preconditioning plus L-arginine hearts (IP+ARG, n=6) received 2 min of ischaemic preconditioning and 5 min reperfusion with 1 mmol/l L-arginine in Krebs Henseleit buffer. Haemodynamic parameters and coronary flow were recorded continuously. Nitrites and nitrates (NOx) were measured 5 and 60 min after reperfusion, and infarct size was also determined. RESULTS: In the IP+ARG group, significant amelioration and preservation of left ventricular peak developed pressure and coronary flow was observed compared with the GI, IP, ARG and IP+L NAME groups. Infarct size in the IP+ARG group was reduced significantly compared with that in the GI, IP, ARG and IP+L-NAME groups. Significant preservation of NOx was observed during reperfusion in the IP+ARG group compared with the GI group. CONCLUSIONS: Inhibition of NO synthase with L-NAME had little impact on ischaemic preconditioning, suggesting that endogenous NO is not a major mediator of ischaemic preconditioning. Nevertheless, enhancement of the effects of ischaemic preconditioning can be achieved with L-arginine, a precursor of NO, improving post-ischaemic functional recovery and infarct size in the isolated rat heart. PMID- 11404146 TI - Pretreatment surgical lymph node staging predicts results of trimodality therapy in esophageal cancer. AB - OBJECTIVE: Prediction of responders to induction therapy in esophageal cancer (EC) patients is important. In this study, we evaluated the role of thoracoscopic/laparoscopic (Ts/Ls) staging in prediction of treatment response and survival in EC patients with trimodality treatment. METHODS: Retrospective study of EC patients who had undergone Ts/Ls staging and received trimodality treatment at the University of Maryland Medical Center and the Baltimore Veterans Administration Hospitals from July, 1991 to December, 1999. Preoperative therapy consisted of concurrent chemotherapy (5-FU + cisplatinum) and radiotherapy. RESULTS: Forty-four EC patients who underwent pretreatment Ts/Ls staging during the study period were able to complete concurrent chemoradiotherapy followed by surgical resection. There were 36 men and 8 women aged 40 to 77 (median age 62). Twenty-seven (61.4%) patients were found to have lymph node metastasis by surgical staging. Fourteen patients (31.8%) had a pathologic complete response. Patients with positive lymph nodes had a lower response rate than those with negative lymph nodes (14.8% vs. 58.8%, P=0.006). Other clinicopathologic features including gender, weight loss, clinical TNM stage, surgical T stage, and histology did not correlate with treatment response. Univariate analysis showed that weight loss and treatment response were important prognostic factors for disease-free survival (P=0.01 and P=0.02, respectively). Histology, surgical N stage and surgical TNM stage appeared to be associated with prognosis (P=0.067 0.097). Multivariate analysis revealed that only surgical N status and weight loss were significant prognostic factors (P=0.05, and P=0.006, respectively). CONCLUSIONS: Surgical Ts/Ls staging provides accurate evaluation of tumor spread in EC patients. Pretreatment N status was the single most important predictor of response to induction treatment as well as a reliable prognosticator of survival. PMID- 11404147 TI - Prognostic evaluation for squamous cell carcinomas of the lower thoracic esophagus treated with three-field lymph node dissection. AB - OBJECTIVE: The efficacy of esophagectomy with three-field lymph node dissection in surgical treatment for patients with squamous cell carcinomas of the lower thoracic esophagus remains controversial. This report documents the outcomes of this surgical procedure for a large series. METHODS: From February 1986 to November 1998, 437 patients with squamous cell carcinomas of the thoracic esophagus underwent transthoracic esophagectomy with three-field lymph node dissection. One hundred and sixteen of these had cancer of the lower thoracic esophagus. To avoid the influence of adjuvant therapy on survival, 20 who also received radiation and/or chemotherapy were excluded, leaving 96 patients who were retrospectively analyzed. RESULTS: The operative morbidity, and 30-day and in-hospital mortality rates were 62, 0, and 3%, respectively. The overall 1-, 3-, and 5-year survival rates were 89, 65, and 59%, with a median survival of 76 months. In those with lymph node metastases (66% of cases), the values were 87, 56, and 48%, as compared with 94, 84, and 79%, respectively (P=0.005) for patients without lymph node metastasis. Factors significantly influencing the overall survival rates were patient age (> or = 65 vs. <65), clinical N status (cN1 vs. cN0), clinical M status (cM1 vs. cM0), longitudinal tumor length of resected specimen (> or =5 vs. <5 cm), pathologic T status (pT3 vs. pT1, 2), pathologic N status (pN1 vs. pN0), lymphatic invasion (positive vs. negative), vascular invasion (positive vs. negative) and intramural metastasis (present vs. absent). Independent prognostic factors for survival determined by multivariate analysis were pathologic T status (P=0.02), pathologic N status (P=0.03), and presence of intramural metastasis (P=0.04). Additional pathologic M1 status, cervical or celiac lymph node metastasis, was without significant influence. CONCLUSIONS: Patients with pathologic T3 tumors with both pathologic N1 status and the presence of intramural metastasis in the lower thoracic esophagus had a poor prognosis. Cervical or celiac lymph node metastasis in patients with carcinomas of the lower thoracic esophagus should be distinguished from pathologic M1 status in the UICC-TNM staging system. PMID- 11404148 TI - Ten year experience with induction therapy in locally advanced non-small cell lung cancer (NSCLC): is clinical re-staging predictive of pathological staging? AB - OBJECTIVE: To verify if in our experience with 'induction therapy' in non-small cell lung cancer (NSCLC) the clinical re-staging is really predictive of pathological staging. MATERIALS AND METHODS: From January 1990 to February 2000, 136 patients with locally advanced NSCLC underwent a protocol of induction therapy according to three different treatment plans: Carboplatin + radiotherapy- study A; Cisplatin + 5-Fluorouracil + radiotherapy--study B; Gemcitabine + radiotherapy--study C. RESULTS: Clinical re-staging showed in the patients enrolled in study A a clinical Complete Response rate (cCR) of 2.3%; a clinical Partial Response rate (cPR) of 50%; a clinical Stable Disease (cSD) rate of 44.3%; a clinical Disease Progression (cDP) rate of 3.4%. In study B, cCR was 0%; cPR: 71.4%; cSD 10.7%; cDP: 17.9%. In study C, cCR was 0%; cPR: 23.5%; cSD: 11.8%; cDP: 64.7%. After clinical re-staging, 76 patients (47 group A; 23 group B; 6 group C) were judged to be resectable and underwent a surgical operation. Pathological staging showed no tumour in eight patients (10.5%; 8/76) (three in study A, four in study B, one in study C) and microscopic neoplastic remnants in seven (9.2%; 7/76). Thirty-nine patients were pN0. Overall downstaging rate in the operated patients was 51%. No precise correlation was found among clinical re staging and pathological staging. We had two cCRs and eight pCRs, and all of these pCRs had been re-staged as cPR except in one case (cSD). In seven cases, where only microscopic remnants have been found, six had been clinically restaged as cPR and one as cSD. CONCLUSIONS: Our experience confirmed how often the clinical re-staging data are unreal. Accordingly surgery should be indicated in any case where an induction therapy has been administered, if it is reasonably possible. PMID- 11404149 TI - Surgical management of metachronous bronchial carcinoma. AB - OBJECTIVE: To assess the results of surgery for the treatment of metachronous bronchial carcinoma. METHODS: From 1985 to 1999, 38 patients were operated on for a metachronous lung carcinoma, accordingly to the criteria of Martini. All tumors were staged using the new International Classification System revised in 1997. RESULTS: Diagnosis of the second cancer was done at radiological follow-up in 30 asymptomatic patients. Seventeen metachronous locations were ipsilateral. Histology of the metachronous lesion was the same as that of the first tumour in 23 patients (60%). The first resection was a lobectomy (n=35), a pneumonectomy (n=2) and a carinal resection (n=1). The second one was a wedge resection (n=7), a segmentectomy (n=3), a lingulectomy (n=2), a lobectomy (n=9), a bilobectomy (n=1), and a pneumonectomy (n=16). There were five in-hospital deaths (13%). Completion pneumonectomy was performed in 15 patients, with one postoperative death (7%). The overall estimated 5 and 10-years actuarial survival rates from the treatment of the first cancer were 70 and 47% respectively. The 5-year survival rate after the treatment of the second cancer was 32% (median survival: 31 months), including the operative mortality. Survival was negatively affected by a resection interval of less than 2 years and the performance of atypical lung sparing pulmonary resection for the treatment of the second cancer. CONCLUSIONS: Good long-term results are achievable by the means of a second pulmonary resection in selected patients with metachronous lung cancer. Optimal cancer operations should be applied whenever functionally possible. PMID- 11404150 TI - Expression of nm23-H1 gene product in mediastinal lymph nodes from lung cancer patients. AB - OBJECTIVE: Although numerous studies have shown that nm23-H1 gene product expression is inversely related to metastatic potential in some cancers, the expression in lymph nodes has not been studied in detail. An analysis of nm23-H1 gene product expression in mediastinal lymph nodes from lung cancer patients is reported. METHODS: One hundred and thirty-four, randomly selected lymph nodes (63 with positive pathological lymph node status) from 39 surgically treated lung cancer patients were examined. Expression of nm23-H1 gene product was determined using specific monoclonal antibodies. Metastatic cancer cells were highlighted using anti-cytokeratin antibody. RESULTS: Expression of nm23-H1 gene product in patients with less and more than 50% nodes-positive was 12/23 (52.2%) and 15/16 (93.8%) cases, respectively. Immunohistochemical studies with cytokeratin revealed micrometastasis in 6/39 (15.4%) patients and 9/71 (12.7%) nodes previously reported as cancer negative. Expression of nm23-H1 gene product in micrometastasis and metastasis-positive nodes was 5/9 (55.6%) and 55/63 (87.3%), respectively. We also found nm23-H1 gene product expression in germinal center cells. However, we found no relationship between expression of nm23-H1 gene product in germinal center cells and extent of metastasis. CONCLUSIONS: Our study demonstrates a positive relationship between expression of nm23-H1 gene product and extent of metastasis in mediastinal lymph nodes from lung cancer patients. Our data for normal germinal center cells suggests that nm23-H1 gene product expression does not play a specific biological role in suppressing tumor metastasis in lung cancer. PMID- 11404151 TI - Survival, disease-free interval, and associated tumor features in patients with colon/rectal carcinomas and their resected intra-pulmonary metastases. AB - OBJECTIVE: Colon/rectum cancer often presents with intrapulmonary metastases. Surgical resection can be performed in a selected group of patients. In this study, the search for possible prognostic factors of patients with primary colon/rectum cancer and lung metastases was performed. METHODS: Medical records of 110 patients operated on pulmonary metastases of primary colon/rectum cancer were reviewed. The clinical parameters include age, sex, pTNM/UICC stage, grading, localization, surgical and adjuvant therapy of the primary cancer. The number, maximum diameter and total intra-thoracic resected tumor-mass ('load'), the pre-thoracotomy serum carcinoembryonic antigen (CEA) levels, localization of the metastases (uni- vs. bilateral), the presence of hilar/mediastinal tumor infiltrated lymph nodes, the surgical procedure and performed therapy schemes of lung metastases were recorded. RESULTS: The cumulated 5- and 10-year total survival after diagnosis of the primary carcinomas was estimated to 71 and 33.7%, respectively. After resection of the pulmonary metastases, the 3- and 5-year post thoracotomy survival measured 57 and 32.6%, respectively. The median time interval between diagnosis of the primary cancer and thoracotomy (disease free interval (DFI)) was found to be 35 months. A non-negligible percentage of patients (15.4%) displayed limited tumor stages of the primary cancer (pT1/2, pN0). The median diameter of the largest metastasis measured 28 mm, and the median resected intrathoracic tumor-load was calculated to 11.4 cm(3). In only 8 patients hilar or mediastinal tumor-involved lymph nodes were found. A potentially curative resection of lung metastases was recorded in 96 patients. The overall survival was significantly correlated with the DFI and the number of intrapulmonary metastases. The DFI correlated significantly with the tumor load and the number of metastases; the post-thoracotomy survival with the number of metastases, tumor-load and pre-thoracotomy serum CEA level. Treatment, stage and grade of the primary cancer, occurrence of liver metastases and local recurrences, mode of treatment of metastases and postoperative residual stage had no significant correlation with either total nor post-thoracotomy survival. CONCLUSIONS: Pulmonary metastases occur even in patients with limited tumor stages of primary colon/rectum cancer. DFI is the major parameter to estimate the total survival of patients with lung metastases. The survival after thoracotomy depends on the number of metastases, the intrapulmonary tumor load and the presence of elevated serum CEA level prior to thoracotomy. PMID- 11404152 TI - Giant hydatid lung cysts. AB - OBJECTIVE: In this clinical retrospective study, we aimed to evaluate giant hydatid lung cyst cases as a different clinical entity that recorded in last 10 years in our clinic. METHODS: Between February 1990 and May 2000, a total of 305 hydatid lung cyst cases from patients that had been operated were reviewed, and 67 (21.9%) cysts with more than 10 cm in diameters of them were regarded as a giant hydatid lung cyst. Further investigations were made with respect to different factors. RESULTS: Thirty-six (54%) cases were male and 31 (46%) were female. The ages ranged between 5 and 54 (mean 21.6) years. The most common symptoms recorded were; cough (68%), thoracic pain (55%) and dyspnea (52%). Cyst sizes were ranged between 10 and 22 cm (mean 13.4) in diameters. Forty-two (62%) of them were in the right, 22 (33%) were in the left hemithorax, and three (5%) were located bilaterally. Cystotomy or cystectomy and capitonnage was the most frequent applied operation procedure (71%). Resection was performed in nine (13%) cases. Thirteen (19%) cases had air leakage more than 10 days in which five (7%) of them empyema occurred postoperatively. One case died due to respiratory failure in fourth postoperative day. The postoperative hospital stay ranged between 6 and 43 (mean 10.5) days. No recurrence was recorded in 1-5 years of a follow-up period. CONCLUSIONS: Giant hydatid lung cysts must be regarded as a different clinical entity because of their early occurrence, having more serious symptoms, with frequent operative complications, and they need prolonged care with higher cost effects. PMID- 11404153 TI - Surgical treatment of bilateral hydatid disease of the lung. AB - OBJECTIVE: To evaluate the various tactics and approaches in the surgical treatment of bilateral pulmonary hydatidosis (BPH). MATERIALS AND METHODS: Between 1969 and 2000, a total of 127 BPH patients underwent surgery. The operative techniques of choice were parenchyma-preserved methods. Up to 1988, two stage operations via thoracotomies were performed on 30 patients. One-stage operations with successive thoracotomies were carried out on two patients. Since 1988, only four patients underwent two-stage operations. One-stage surgery was carried out on 91 patients: 82 via median sternotomy (MS), one via clamshell incision, four through successive thoracotomies and three through video-assisted thoracic surgery (VATS) and mini-thoracotomies. One-stage bilateral lower lobectomies via MS were performed on one patient. In 11 cases, the concomitant dome localized liver cysts were extirpated via right phrenotomy during MS. Sterno laparotomy was performed on 11 patients: for associated hepatic (seven), and hepatic and spleen cystectomies (four). In eight cases, abdominal echinococcosis was operated on a second stage, and in one case, a complicated hepatic cyst was extirpated on a first stage. RESULTS: No intraoperative deaths occurred. The postoperative mortality rate was 0.78%; one patient died of pulmonary embolism. No fatal complications have appeared in eight cases (skin suppuration, residual pleural cavity and atelectasis). Adult respiratory distress syndrome was successfully treated in one case after MS. The long-term postoperative results are considered very good, with no recurrences observed. CONCLUSIONS: One-stage surgery is superior to a classic two-stage approach as it decreases the morbidity, hospital stay and costs. MS is an excellent approach, but in some cases, VATS mini-thoracotomies could be indicated. PMID- 11404154 TI - Internal comparative audit in a thoracic surgery unit using the physiological and operative severity score for the enumeration of mortality and morbidity (POSSUM). AB - OBJECTIVE: The aim of the study was to use the physiological and operative severity score for the enumeration of mortality and morbidity (POSSUM) to assess the performance of our thoracic surgery unit during two successive periods of activity. METHODS: From 1992 through 1997, 801 candidates for thoracic procedures at our institution were considered for the study. After validation of the logistic regression model, including the POSSUM score, observed and POSSUM predicted morbidities were compared within two groups of patients divided by year of operation (group 1: 1992-1994, n=362; group 2: 1995-1997, n=439) by means of the z-test for the comparison of a proportion with an expected value. RESULTS: The POSSUM-predicted morbidity was significantly lower than the observed one in the first period of activity of our unit (19.6 vs. 24.3%, respectively; z-test for the comparison of a proportion with an expected value, 2.25; P=0.01), whereas no difference was found in the second period (20.5 vs. 19.1%, respectively; z test for the comparison of a proportion with an expected value, -0.71; P=0.76). CONCLUSIONS: The result suggests a worse-than-expected performance of our unit in the first period of activity, showing that POSSUM can be reliably applied as an instrument of internal comparative audit in a thoracic surgery unit. PMID- 11404155 TI - Screening of a stentless valve with electron beamed tomograpy 5 years after operation. PMID- 11404156 TI - A different surgical approach to the treatment for early stage bronchopleural fistulas. PMID- 11404157 TI - Giant chondroma of the anterior chest wall. PMID- 11404158 TI - Posterior wall laceration of the thoracic trachea: the transcervical transtracheal approach. AB - A large membranous wall laceration of the thoracic trachea was surgically treated. The surgical approach consists on a low collar incision followed by a longitudinal tracheotomy. The membranous tear was repaired with a running suture and tracheotomy sutured with interrupted crossed stitches. The procedure was effective and endoscopic follow-up showed a perfect healing process with no signs of tracheal stenosis. This new technique proved to be a reliable, quick and safe procedure, which allows to repair membranous lacerations as far as the carina, avoiding thoracotomy. PMID- 11404159 TI - Simultaneous coronary arterial bypass surgery using mini-sternotomy and off-pump methods and abdominal aneurysm repair. AB - A technique of combined minimally invasive coronary artery surgery and abdominal aneurysm repair is described. A mini-sternotomy and off-pump coronary artery bypasses to the left descending branch and right coronary arteries are conducted before abdominal aneurysm repair in a simultaneous operation. PMID- 11404160 TI - Carotid artery pseudoaneurysm in Behcet's disease. AB - We report a case of carotid artery pseudoaneurysm occurring in a patient with Behcet's disease for the purpose of discussing approach to this unusual complication of Behcet's disease. PMID- 11404161 TI - Constrictive tuberculous pericarditis in a HIV-positive patient. AB - Constrictive pericarditis is a relatively rare clinical manifestation nowadays. We present the case of an HIV-positive patient with constrictive calcified pericarditis due to an infection with Mycobacterium tuberculosis. Pericardectomy was performed. The therapeutical approach is discussed and the literature is reviewed. PMID- 11404162 TI - Congenital aneurysm of the right atrium. AB - Congenital aneurysm of the right atrium is described in a 1-year-old girl who presented with cardiomegaly and symptoms of paroxysmal supraventricular tachycardia. Echocardiography and cineangiography both established the definitive diagnosis and surgical resection was successful. The rarity of this condition is pointed out and its main features outlined. PMID- 11404163 TI - Postinfarction ventricular septal perforation repair with endoventricular circular patch plasty using double patches and gelatin-resorcinol-formaldehyde biological glue. AB - We describe a technique for repairing the ventricular septal perforation 10 days post acute anteroseptal myocardial infarction using the modified infarction exclusion method. The repair involves endoventricular circular patch plasty and application of gelatin-resorcinol-formaldehyde biological glue in the space between a Teflon felt patch on the infarcted septum and a bovine pericardial patch in the left ventricular cavity. Its use in a patient resulted in an almost normal shaped interventricular septum and left ventricular cavity as well as normal left ventricular function. PMID- 11404164 TI - Christiaan Barnard and his contributions to heart transplantation. AB - Christiaan (Chris) Barnard was born in South Africa in 1922 and qualified in medicine at the University of Cape Town in 1946. Following surgical training in Cape Town and Minneapolis, Barnard established a successful open heart surgery program at Groote Schuur Hospital. In 1967, he led the team that performed the world's first human-to-human heart transplant. Although his first patient survived only 18 days, 4 of his first 10 patients survived for more than 1 year, 2 living for 13 and 23 years, respectively. With his junior colleague, Jaques Losman, Barnard then developed the operation of heterotopic heart transplantation. In 1981, his group was the first to successfully transport donor hearts using a hypothermic perfusion storage device. Several studies on the hemodynamic and metabolic sequelae of brain death were carried out in his department. Barnard retired from the University of Cape Town in 1983 at the age of 61. Now in his 79th year, he continues to pursue his interest in writing for the public. In the words of one of his former colleagues, he is a "surgical visionary and simply the most unforgettable character of the second generation of cardiac surgeons." PMID- 11404165 TI - Safety and efficacy of pravastatin therapy for the prevention of hyperlipidemia in pediatric and adolescent cardiac transplant recipients. AB - BACKGROUND: Hyperlipidemia is common after cardiac transplantation and it is a risk factor for post-transplantation coronary artery disease. Immunosuppression with corticosteroids and cyclosporine has been associated with hyperlipidemia. Pravastatin, a HMG-CoA reductase inhibitor, has been shown to be effective and safe for cholesterol reduction in adult heart transplant recipients. To our knowledge the safety and efficacy of pravastatin therapy in pediatric and adolescent heart transplant populations have not been previously analyzed. Therefore, we evaluated lipid profiles, liver transaminases, rejection data, and possible side effects in pediatric and adolescent cardiac transplant recipients treated with pravastatin. METHODS: The study group consisted of 40 cardiac transplant recipients 10 to 21 years old (mean age 16.9 years). Twenty-two patients received pravastatin in addition to an immunosuppressive regimen of either cyclosporine or tacrolimus, azathioprine or mycophenolate mofetil, and prednisone. Serial determinations of total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein, and triglycerides were available for all pravastatin-treated patients. Pre-treatment lipid values and hepatic transaminases were compared with those measured after therapy with pravastatin. Comparison of pravastatin-induced lipid reduction between groups treated with cyclosporine vs tacrolimus was also made. RESULTS: Patients receiving pravastatin experienced a mean 32 mg/dl decrease in TC (p < 0.005) and a mean 31 mg/dl decrease in LDL (p < 0.005), regardless of their immunosuppressive regimen. No statistical differences occurred in the magnitude of mean lipid reduction induced by pravastatin between the groups treated with cyclosporine vs tacrolimus. No significant changes in hepatic transaminase levels were noted, and no clinical evidence of pravastatin-induced myositis occurred in any subjects. CONCLUSION: Pravastatin therapy is effective and safe when used in pediatric and adolescent cardiac transplant recipients. Although the pravastatin-induced reduction in TC and LDL was more pronounced in patients receiving cyclosporine, the reduction was not statistically different from that in the tacrolimus group. No evidence of hepatic dysfunction or rhabdomyolysis in patients treated with pravastatin was noted. Long-term studies are required to evaluate the effect of pravastatin therapy on the incidence of accelerated coronary atherosclerosis in this population. PMID- 11404166 TI - Airway growth after pediatric lung transplantation. AB - BACKGROUND: Lung transplantation (LT) has been successfully offered to pediatric patients. Very little is known about the growth of the transplanted lung, especially in the infant population. Computerized tomography (CT) scanning is a simple method for studying pediatric patients who have undergone LT. We evaluated the use of CT scans to assess airway growth after pediatric LT, compare airway diameter indexed to somatic growth between LT patients and normals, and compare the growth of pre-anastomotic and post-anastomotic airways indexed to somatic growth in pediatric LT patients. METHODS: We reviewed CT scans on all pediatric patients who underwent primary LT before their fifteenth birthday between January 1995 and September 1998. Uniform measurements of diameter were made in pre anastomotic (trachea, and proximal right and left bronchi) and post-anastomotic (distal right and left bronchi) sites. These measurements were then correlated with height and compared to previously published normal values. RESULTS: Of the 16 patients who underwent LT during the study period, 11 had at least 2 sequential CT scans (LT age 3 months to 14 years, median 2 years). Thirty-one CT scans were reviewed. Inter-observer variability was within 1 standard deviation (2 mm) in 93% of the measurements and inter-observer reliability was 0.91 by analysis of variance. Tracheal transverse diameter plotted against body height (slope 0.0072, correlation coefficient 0.88) was virtually identical to previously published norms. A similar relationship between airway diameter and height was observed in pre-anastomotic and post-anastomotic segments. CONCLUSION: CT scanning is a reliable method for assessing airway growth in pediatric LT recipients. Tracheal growth in pediatric LT recipients is similar to that of normal children. Post-anastomotic large airways grow similarly to native, pre anastomotic airways. PMID- 11404167 TI - The effect of cytokine gene polymorphisms on pediatric heart allograft outcome. AB - BACKGROUND: Cytokines play a major role in the inflammatory and immune responses that mediate allograft outcome. Several studies have shown that the production of cytokines varies among individuals and these variations are determined by genetic polymorphisms, most commonly within the regulatory region of the cytokine gene. The aim of this study was to assess the effect of these allelic variations on acute rejection after pediatric heart transplantation. METHODS: We performed cytokine genotyping using polymerase chain reaction-sequence specific primers in 93 pediatric heart transplant recipients and 29 heart donors for the following functional polymorphisms: tumor necrosis factor-alpha (TNF-alpha) (-308), interleukin (IL)-10 (-1082, -819, and -592), TGF-beta1 (codon 10 and 25), IL-6 ( 174), and interferon-gamma (INF-gamma) (+874). The distribution of polymorphisms in this population did not differ from published controls. The patients were classified as either non-rejecters (0 or 1 episode) or rejecters (> 1 episode) based on the number of biopsy proven rejection episodes in the first year after transplantation. RESULTS: Forty-two of the 69 TNF-alpha patients (61%) in the low producer group were non-rejecters, while 9 of the 24 (37.5%) with high TNF-alpha were non-rejecters (p = 0.047). In contrast, IL-10 genotype showed the opposite finding. Forty-two of the 66 patients (64%) in the high and intermediate IL-10 group were non-rejecters, while 9 of the 26 (35%) in the low IL-10 group were non rejecters (p = 0.011). The combination of low TNF-alpha with a high or intermediate IL-10 genotype was associated with the lowest risk of rejection (34/49 or 69% non-rejecters). Neither the distribution of the IL-6, INF-gamma, and TGF-beta1 genotype in recipients nor the donor genotype showed any association with acute rejection. CONCLUSION: Genetic polymorphisms that have been associated with low TNF-alpha and high IL-10 production are associated with a lower number of acute rejection episodes after pediatric heart transplantation. PMID- 11404168 TI - Early intervention after severe oxygenation index elevation improves survival following lung transplantation. AB - BACKGROUND: Reperfusion injury and technical problems following lung transplantation may result in life-threatening pulmonary dysfunction that requires intervention with either extracorporeal membrane oxygenation or reoperation. Early intervention in these patients could prevent complications associated with delayed or emergent intervention and may improve survival. The oxygenation index [(mean airway pressure x percent of inspired oxygen)/partial pressure of arterial oxygen] provides a rapid assessment of pulmonary function in the critical phase of reperfusion. Our hypothesis was that the oxygenation index could be used as an early predictor for severe respiratory failure requiring acute intervention. METHODS: Analysis of 136 consecutive lung transplant operations revealed 18 patients (reperfusion injury in 16 and technical complications in 2) with an oxygen index of > or = 30. Of those patients with reperfusion injury, 9 had fibrotic lung disease, 4 had obstructive lung disease, and 3 had primary pulmonary hypertension. RESULTS: Patients undergoing transplantation for fibrotic lung diseases were more likely to develop severe reperfusion injury (oxygenation index > or = 30) compared to patients with obstructive lung diseases (9 of 42 or 21% vs 4 or 80 or 5%, p = 0.005). The 5 patients with early intervention (< or = 2 hours) after an oxygenation index elevation above 30 had significantly improved survival compared to the 13 with no or late intervention (80% vs 15% survival, p = 0.02). CONCLUSION: Oxygenation index elevation > or = 30 following lung transplantation is an early predictor of severe respiratory failure requiring acute intervention. Early intervention in these patients improves survival. PMID- 11404169 TI - The Brazilian experience with heart transplantation: a multicenter report. AB - BACKGROUND: The results of heart transplantation in developing countries are influenced by the high incidence of marginal donors and the large number of recipients with characteristics of alternative list. The purpose of this multicenter report was to determine the rate of survival after heart transplantation in a developing country. Also we studied the causes of death, the results based on the year of transplant, the influence of gender and age, the numbers of transplants per year, and the etiology of the cardiomyopathy causing the heart failure. METHODS: We studied 792 (632 male) patients who underwent orthotopic heart transplantation at 16 centers. The mean age of the patients was 42 +/- 16 years. Etiology included idiopathic dilated cardiomyopathy in 407 patients, ischemia in 196 patients, Chagas disease in 117 patients, and various other in 72 patients. Cyclosporine was the cornerstone of the immunosuppression administered. RESULTS: Survival for the entire population at 3 months and 1, 4, 8, and 12 years was 72%, 66%, 54%, 40%, and 27%, respectively. There was an improvement in survival from 1991 to 1995 compared with before 1991. Age and gender did not influence the results. Unexpected early mortality was observed, but the late results were satisfactory. The most prevalent causes of death were infection in 23%, acute graft failure in 19%, and rejection in 18%. CONCLUSIONS: Heart transplantation has become feasible in developing countries and the survival rate has improved without the influence of gender and age recipients. A chagasic etiology was found to be the third-leading indication for heart transplantation. The impact of increment of donors with appropriate care for reduction of marginal donors, perhaps associated with better recipient selection and postoperative care, should be investigated for improving early results. PMID- 11404170 TI - Development of anti-major histocompatibility complex class I or II antibodies following left ventricular assist device implantation: effects on subsequent allograft rejection and survival. AB - BACKGROUND: Previous reports have indicated that antibodies to HLA class I or II antigens develop in approximately 60% of patients following left ventricular assist device (LVAD) implantation, subsequent rates of allograft rejection are higher, and survival is adversely affected. METHODS: We performed an analysis of the incidence of antibody development to HLA class I or II antigens by panel reactive antibody (PRA) screening following implantation of the HeartMate LVAD in 38 patients from October 1, 1996 to March 1, 2000 (6 LVAD deaths excluded from study). The occurrence of vascular or cellular rejection of International Society of Heart and Lung Transplantation grade > or = 3A, as determined by endomyocardial biopsy following heart transplantation (HTX), were compared for patients with (n = 32, LVAD group) or without (n = 68, control group) preoperative LVAD support. RESULTS: After LVAD implantation, 9 patients (28%) in the LVAD group developed IgG antibodies to class I (n = 3), class II (n = 5), or both antigens (n = 1) with PRA > 10%. The remaining 23 patients (72%) had either no detectable IgG antibody development or IgG antibody development with PRA < 10%. At the time of HTX, only 4 patients in the LVAD group had persistent PRA > 10%. Only 3 patients (4%) in the control group had PRA > 10% at the time of HTX. The incidence of patients free from rejection at 6 and 12 months was 62% and 44% for the control group, and 49%, and 40% for the LVAD group, respectively (p not significant). The mean linearized rate plus or minus standard deviation of allograft rejection from 0 to 6 months and 7 to 12 months was 0.13 +/- 0.21 and 0.09 +/- 0.14 episodes a month, respectively, for patients with no LVAD support, and 0.17 +/-.25 and 0.06 +/- 0.1 episodes a month, respectively, for those with LVAD support (p = not significant). Post-transplantation survival at 1 and 2 years was 90% and 90%, respectively, for the control group, and 97% and 92%, respectively, for the LVAD group (p not significant). CONCLUSION: Patients with LVAD support before HTX do not appear to be at increased risk for significant allograft rejection in the first year or for death within the first 2 years after transplantation. PMID- 11404171 TI - Synthetic membrane neo-pericardium facilitates total artificial heart explantation. AB - BACKGROUND: In the past, explantation of the Cardio West total artificial heart (TAH) has been technically challenging because of the presence of dense adhesions and extremely thickened pericardium. To prevent this, we constructed a synthetic neo-pericardium in 14 patients. METHODS: Using expanded polytetrafluoroethylene (e-PTFE) membrane, we constructed a pericardium within the pericardium, or "neo pericardium," completely covering the Cardio West TAH separating the native atria from the native pericardium, and wrapping the ascending aorta from the outflow conduit distally for about 5 to 7 cm. RESULTS: Of the 14 patients, 9 were transplanted and could be evaluated, 3 died on device support, and 2 are currently on device support. In each case, we attained faster (by 25 minutes) and easier reentry through the sternum. Surgical planes around the aorta, over the right and left atria, and throughout the pericardial space became apparent immediately after e-PTFE membrane removal. The pericardium and related tissues although slightly thickened (<2 mm) were pliable compared with our previous 36 patients, with very thick adherent pericardium over the device and native atria. CONCLUSIONS: The plastic materials forming the ventricular housing and drivelines of the Cardio West TAH and the Dacron outflow conduits have in the past caused profound local inflammatory reactions, resulting in extremely dense adhesions and thickened adherent pericardium. Using e-PTFE membrane to fashion a complete neo pericardium and to wrap the ascending aorta at the time of Cardio West implantation dramatically reduces adhesions and pericardial thickening and facilitates explantation. PMID- 11404172 TI - Improvement in quality of life outcomes 2 weeks after left ventricular assist device implantation. AB - BACKGROUND: The successful use of left ventricular assist devices (LVADs) as a bridge to heart transplantation has prompted our examination of quality of life (QOL) outcomes. The purposes of this study are to describe QOL in patients 1 to 2 weeks after LVAD implantation and to compare QOL in a smaller cohort of patients from before to 1 to 2 weeks after surgery. METHODS: Data were collected from a convenience sample of 81 patients who completed booklets of questionnaires that measure domains of QOL 1 to 2 weeks after LVAD insertion and from 30 of 81 patients who completed booklets at both the pre-implantation and post implantation periods. Patients completed booklets of 6 to 8 self-reporting instruments, with acceptable reliability and validity. Data were analyzed using descriptive and comparative statistics (chi-square, Mann-Whitney U and Wilcoxon signed ranks tests) with p = 0.01 considered statistically significant. RESULTS: One to 2 weeks after LVAD implantation, patients were quite satisfied with their lives, experienced moderately low amounts of stress, coped well, and perceived themselves as having good health and QOL, low symptom distress, and moderately low functional disability. Patients reported significantly better QOL, more satisfaction with health and functioning, and were significantly less distressed by symptoms from immediately pre-operatively to post-operatively. However, patients reported significantly more self-care disability and more dissatisfaction with socioeconomic areas of life from before to immediately after surgery. Psychological distress was low and did not change with time. CONCLUSION: Given that QOL improved from before to after LVAD implantation, our findings provide a springboard for investigation of the impact of LVADs on long-term QOL outcomes. PMID- 11404173 TI - A leukocyte depleting filter reduces endothelial cell dysfunction and improves transplanted canine heart function. AB - BACKGROUND: To date leukocytes have been known to play a major role in reperfusion injury and have directed attention to leukocyte-endothelium interaction. This study was designed to investigate how much graft viability and the coronary microcirculatory function could be preserved by leukocyte depletion (LD) in a model of orthotopic cardiac transplantation. METHODS: The heart in 10 beagle dogs was arrested by introducing a 4 degrees C St. Thomas' cardioplegic solution. They were harvested, immersed in the cold saline for 3 hours, and then orthotopically transplanted. Five recipients underwent LD (LD group) at reperfusion with the use of a Pall BC1B leukocyte depleting filter inserted into the cardiopulmonary bypass (CPB) circuit. The other 5 dogs without filtration served as a control group. RESULTS: Leukocytes were about 80% filtrated and neutrophils were also 85% filtrated during the first 30 minutes of reperfusion in the LD group. A high level of adenosine triphosphate was maintained after transplantation in the LD group. The polymorphonuclear elastase level was significantly lower in the LD group. The cardiac function assessed by the slopes of the end-systolic pressure volume relation after transplantation was significantly higher in the LD group than in the control group (p < 0.05). The coronary vascular resistance responses to acetylcholine and nitroglycerin after transplantation were preserved significantly better in the LD group than in the control group (p < 0.05). CONCLUSIONS: These results suggest that a leukocyte depleting filter placed in the CPB circuit would prevent leukocyte-mediated endothelial cell injury, improve microcirculation of the myocardium, and lead to excellent graft function. PMID- 11404174 TI - Increased matrix metalloproteinase 9 activity and mRNA expression in lung ischemia-reperfusion injury. AB - OBJECTIVES: In lung ischemia-reperfusion injury, neutrophil migration from the vasculature to the interstitial spaces plays a major role in tissue injury. Degradation of the basement membrane, which is composed of extracellular matrix (ECM) molecules, is necessary for neutrophil migration. Matrix metalloproteinases (MMPs) might play a role in ECM degradation in lung ischemia-reperfusion injury. We evaluated the changes in the activity of MMP-2 and MMP-9, and tissue inhibitor of metalloproteinase 1 (TIMP-1) gene expressions using rat lung transplantation models. METHODS: We divided animals into 4 groups. Groups I and II served as control groups with intact lungs (Group I) and 24-hour cold-preserved lungs (Group II). Groups III and IV received lung grafts after 24-hour cold preservation. The recipient animals were sacrificed 1 hour (Group III) or 24 hours (Group IV) after transplantation. We evaluated lung injury histologically. We assessed MMP activity using zymography. We assessed MMP-2, MMP-9, and TIMP-1 gene expression using biplex reverse transcriptase-polymerase chain reaction method. RESULTS: In Groups III and IV, we noted severe ischemia-reperfusion injury. We noted no significant difference in enzyme activity and gene expression of MMP-2 between Groups I and IV. The MMP-9 activity and gene expression were low during ischemia and increased on reperfusion. TIMP-1 gene expression was low during ischemia and at the early phase of reperfusion, and showed a dramatic increase at the late phase of reperfusion. CONCLUSIONS: Matrix metalloproteinase 9, but not MMP-2, may play an important role in ischemia-reperfusion injury. TIMP 1 increases at the late phase of reperfusion and may compensate for the activity of MMP-9. PMID- 11404175 TI - Transmyocardial laser revascularization in ischemic cardiomyopathy. AB - BACKGROUND: In ischemic cardiomyopathy (left ventricular ejection fraction [LVEF] < or = 30%), myocardial revascularization by coronary artery surgery has better results than heart transplantation, provided there is sufficient ischemic but viable myocardium. The mode of action of transmyocardial laser revascularization (TMR) is still being debated, but if the procedure induces improved myocardial perfusion it could be a "bridge," or alternative, to heart transplantation. METHODS: We retrospectively analyzed 194 patients, who underwent TMR between July 1997 and October 1999. Patients with TMR as an adjunct to coronary artery surgery (n = 30) and those who did not provide written consent to the procedure (n = 8) were excluded; 126 patients had normal or moderately reduced left ventricular function, and 30 patients with ischemic cardiomyopathy (LVEF < or = 30%) were included. RESULTS: After 12 months, the Canadian Cardiovascular Society (CCS) score dropped significantly from 3.6 (3 to 4) to 2.4 (1 to 4) and maximum work load increased significantly from 58 W (25 to 100 W) to 73 W (25 to 120 W). However, thallium score and LVEF did not improve significantly (27% [15% to 30%] to 32% [15% to 45%]). Prior to the TMR procedure, all 30 patients had a "low risk" or "medium risk" of death according to the Aaronson classification. The 30 day, 1-year, and 2-year survival rates were 83%, 50%, and 47%, respectively. CONCLUSIONS: We conclude that TMR in ischemic cardiomyopathy (LVEF < or = 30%) has a perioperative risk comparable to that for heart transplantation, but there is no improvement of myocardial performance or life expectancy. Therefore, TMR cannot be regarded as a "bridge," or alternative, to transplantation. However, in individual cases with contraindications for transplantation the anti-anginal effect may justify use of the procedure. PMID- 11404176 TI - Eosinophilic infiltrates in a pulmonary allograft: a case and review of the literature. AB - An unusual case of peribronchial eosinophilic infiltrates associated with peripheral blood eosinophilia in a lung transplant patient is described. The role that eosinophils play in lung allograft rejection is reviewed. Tissue eosinophils have been associated with acute pulmonary allograft rejection. Although, eosinophils in bronchoalveolar lavage fluid (BAL) have been observed in allograft rejection, this relationship is less well defined. The role of eosinophils in the pathophysiology of allograft rejection is unclear. PMID- 11404177 TI - Cardiac transplantation in a patient with hereditary hemochromatosis: role of adjunctive phlebotomy and erythropoietin. AB - We describe the case of a 36-year-old woman with hereditary hemochromatosis (HH) resulting in end-stage cardiomyopathy and treated successfully with orthotopic cardiac transplantation. Before and after transplantation, the patient underwent aggressive treatment with frequent phlebotomy. We used erythropoietin concomitantly to maintain adequate hematocrit to support continued phlebotomy. We believe that aggressive use of phlebotomy provided the patient hemodynamic benefit and hastened the return of endocrine function post-transplantation. We also believe that the patient's history of high-dose vitamin C usage may have accelerated iron deposition in the heart and other vital organs. PMID- 11404178 TI - A technique of cervical aortic graft transplantation in mice. AB - A new method of mouse aortic graft transplantation into carotid artery was developed with cuff technique. By harvesting the descending aorta of the donor using a small Teflon cuff (external diameter 0.6 mm, internal diameter 0.4 mm) and super fine-tip forceps, and modifying the method of mouse heterotopic heart transplantation with cuff technique, donor descending aortic allografts could be interposed in the common carotid artery of recipient mice. Histological analysis demonstrated neither evidence of tissue damage nor intimal thickening in isograft implanted over 100 days. We strongly recommend that this new model of aortic transplantation in mice is a simple and useful technique for vascular transplantation research. PMID- 11404179 TI - Anticarcinogenic actions of melatonin which involve antioxidative processes: comparison with other antioxidants. AB - The complex processes of carcinogenesis often involve oxidative stress. Numerous indicators of oxidative damage are enhanced as the result of the action of carcinogens. Several antioxidants, with different efficacies, protect against oxidative abuse caused by carcinogens. Recently, melatonin (N-acetyl-5 methoxytryptamine) and related indoleamines have attracted attention because of their high antioxidant and anticarcinogenic activity. Some antioxidants, e.g. ascorbic acid, play an ambivalent role in antioxidative defense, since, under specific conditions, they are strongly prooxidant. Among known antioxidants, melatonin has been an often investigated experimental agent in reducing cancer initiation and inhibiting the growth of established tumors. The indoleamine has been shown to protect macromolecules from oxidative mutilation induced by carcinogens. In these studies, a variety of in vitro and in vivo models were used and numerous indices of oxidative damage were evaluated. The protective effects of melatonin and several other indoleamine antioxidants against cellular damage caused by carcinogens make them potential supplements in the treatment or co treatment at several stages of cancer. PMID- 11404180 TI - Regulation of oxygen sensing in peripheral arterial chemoreceptors. AB - The carotid bodies are a small pair of highly vascularized and well perfused organs located at each carotid artery bifurcation, strategically situated to sense oxygen in arterial blood as it leaves the heart. Carotid body glomus cells are identified as the primary oxygen sensors, which respond to changes in blood P(O(2)) within milliseconds. Acute hypoxia causes a rapid increase in carotid sinus nerve (CSN) activity, providing afferent signals to the respiratory center in the brainstem. Glomus cells secrete numerous neurotransmitters that modulate CSN firing rates. This review will discuss major hypotheses that have emerged regarding acute oxygen sensing by glomus cells. In contrast, chronic responses to hypoxia are much slower, involving cytosolic reactions that take place over several minutes and nuclear reactions which occur over several hours. Converging concepts from different areas of research in oxygen sensing cells and tissues (including the carotid body) have been combined to describe molecular and biochemical changes that take place in the carotid body with chronic hypoxia. These include oxygen dependent proteolytic processes in the cytosol and gene transcription in the nucleus. In addition, cellular and nuclear responses to chronic hypoxia will be discussed. PMID- 11404181 TI - DNA oxidatively damaged by chromium(III) and H(2)O(2) is protected by the antioxidants melatonin, N(1)-acetyl-N(2)-formyl-5-methoxykynuramine, resveratrol and uric acid. AB - Chromium (Cr) compounds are widely used industrial chemicals and well known carcinogens. Cr(III) was earlier found to induce oxidative damage as documented by examining the levels of 8-hydroxydeoxyguanosine (8-OH-dG), an index for DNA damage, in isolated calf thymus DNA incubated with CrCl(3) and H(2)O(2). In the present in vitro study, we compared the ability of the free radical scavengers melatonin, N(1)-acetyl-N(2)-formyl-5-methoxykynuramine (AFMK), resveratrol and uric acid to reduce DNA damage induced by Cr(III). Each of these scavengers markedly reduced the DNA damage in a concentration-dependent manner. The concentrations that reduced 8-OH-dG formation by 50% (IC(50)) were 0.10 microM for both resveratrol and melatonin, and 0.27 microM for AFMK. However, the efficacy of the fourth endogenous antioxidant, i.e. uric acid, in terms of its inhibition of DNA damage in the same in vitro system was about 60--150 times less effective than the other scavengers; the IC(50) for uric acid was 15.24 microM. These findings suggest that three of the four antioxidants tested in these studies may have utility in protecting against the environmental pollutant Cr and that the protective effects of these free radical scavengers against Cr(III) induced carcinogenesis may relate to their direct hydroxyl radical scavenging ability. In the present study, the formation of 8-OH-dG was likely due to a Cr(III)-mediated Fenton-type reaction that generates hydroxyl radicals, which in turn damage DNA. Once formed, 8-OH-dG can mutate eventually leading to cancer; thus the implication is that these antioxidants may reduce the incidence of Cr related cancers. PMID- 11404182 TI - Carbohydrate deprivation reduces NADPH-production in fish liver but not in adipose tissue. AB - Little is known about the way in which carnivorous fish such as salmonids mobilise and metabolise dietary carbohydrates, which are essential to lipid metabolism. Thus we have studied changes caused by the absence of dietary carbohydrates to the kinetics and molecular behaviour of the four cellular NADPH production systems [glucose 6-phosphate dehydrogenase (G6PDH); 6-phosphogluconate dehydrogenase (6PGDH); malic enzyme (ME); and isocitrate dehydrogenase NADP dependent (NADP-IDH)] in the liver and adipose tissue of rainbow trout (Oncorhynchus mykiss). We used spectrophotometry to study enzyme kinetics and nucleic acid concentrations, and immunoblot analysis to determine specific protein concentrations. The absence of carbohydrate reduced specific enzyme activity, maximum rate and catalytic efficiency by almost 65% in G6PDH and 6PGDH, by more than 50% in ME, and by almost 25% in NADP-IDH but caused no significant changes in the K(m) values or activity ratios in any of these hepatic enzymes. Molecular analysis clearly showed that this kinetic behaviour reflected concomitant changes in intracellular enzyme concentrations, produced by protein induction/repression processes rather than changes in the activity of pre existing enzymes. We conclude that the absence of carbohydrates significantly reduces intracellular concentrations of G6PDH, ME and NADP-IDH in trout liver in percentages similar to those recorded for enzyme activity. We found no such variations in the concentrations of any of these enzymes in adipose tissue and no change in the levels of their activity, suggesting that the liver and adipose tissues are subject to different regulation systems with regard to carbohydrates and play distinct roles in lipid metabolism. PMID- 11404183 TI - Alterations in interprotein interactions between translation initiation factors assessed by fluorescence resonance energy transfer. AB - Interaction of the translational repressor 4E-BP1 with the mRNA cap binding protein eIF4E plays an important role in the regulation of translation initiation. This interaction is modulated by phosphorylation of 4E-BP1 on at least six residues. However, analysis of the functional importance of the individual phosphorylation sites is complicated by the lack of information about the kinases and phosphatases involved in modulating phosphorylation of each site. The goal of the present study was to establish a system whereby alterations in the interaction of 4E-BP1 with eIF4E could be easily and directly measured. In initial studies, both eIF4E and 4E-BP1 were expressed as recombinant proteins coupled to variants of green fluorescent protein (ECFP and EYFP, respectively). Addition of purified EYFP--4E-BP1 to ECFP--eIF4E caused both a decrease in emission intensity at 480 nm and an increase at 535 nm indicating that protein protein interaction had occurred. The interaction was stoichiometric and was blocked by eIF4G. Phosphorylation of EYFP--4E-BP1 by the mitogen-activated protein kinase ERK2, but not by casein kinase CK-II, also attenuated the interaction. Results using proteins in which the fluorescent protein tag was located at either the N- or C-terminus suggested that, in the protein complex, the N-termini of the two proteins are in close spatial proximity, as are the C termini. Overall, the results demonstrate that fluorescence resonance energy transfer between EYFP--4E-BP1 and ECFP--eIF4E is a valuable tool in directly measuring alterations in the interaction of the two proteins. PMID- 11404184 TI - The interaction of acyl-CoA with acyl-CoA binding protein and carnitine palmitoyltransferase I. AB - The affinity of recombinant rat acyl-CoA binding protein (ACBP) towards acyl-CoAs was investigated using both fluorimetric analysis and isothermal titration microcalorimetry, neither of which requires the physical separation of bound and free ligand for determining the dissociation constants (K(d)). The displacement of 11-(dansylamino)undecanoyl-CoA (DAUDA-CoA) from ACBP yielded binding parameters for the competing acyl-CoAs that compared favourably with those obtained using ultra-sensitive microcalorimetric titration. The K(d) values of ACBP for oleoyl-CoA and docosahexaenoyl-CoA are 0.014 and 0.016 microM, respectively. Under identical experimental conditions, carnitine palmitoyltransferase I (CPT I) of purified rat liver mitochondria has K(d) values of 2.4 and 22.7 microM for oleoyl-CoA and docosahexaenoyl-CoA, respectively. Given that CPT I was not only present at a much lower concentration but also has an appreciably lower affinity for acyl-CoAs than ACBP, it is proposed that CPT I is capable of interacting directly with ACBP-acyl-CoA binary complexes. This is supported by the fact that the enzyme activity correlated with the concentration of ACBP-bound acyl-CoA but not the free acyl-CoA. A transfer of acyl-CoA from ACBP-acyl-CoA binary complexes to CPT I could be a result of the enzyme inducing a conformational alteration in the ACBP leading to the release of acyl-CoA. PMID- 11404185 TI - Effects of adenosine receptor antagonism on protein tyrosine phosphatase in rat skeletal muscle. AB - Earlier studies have shown that whole body adenosine receptor antagonism increases skeletal muscle insulin sensitivity in insulin-resistant Zucker rats. To find which steps in the insulin signaling pathway are influenced by adenosine receptors, muscle from lean and obese Zucker rats, treated for 1 week with the adenosine receptor antagonist, 1,3-dipropyl-8-(4-acrylate)-phenylxanthine (BWA1433), were analyzed. All rats were first anesthetized and injected intravenously (i.v.) with 1 IU of insulin. About 3 min later the gastrocnemius was freeze clamped. Insulin receptors were partially purified on wheat germ agglutinin (WGA) columns and insulin receptor kinase activity measured in control and BWA1433-treated lean and obese Zucker rats. Protein tyrosine phosphatase (PTPase) activity was also analyzed in subcellular fractions, including the cytosolic fraction, a high-speed particulate fraction and the insulin receptor fraction eluted from WGA columns. Administration of BWA1433 increased insulin receptor kinase activity in obese but not lean Zucker rats. PTPase activities were higher in the untreated obese rat muscle particulate fractions than in the lean rat particulate fractions. The BWA1433 administration lowered the PTPase activity of the obese rats but not the lean rats. Although the PTPase activity in WGA eluate fractions containing crude insulin receptors were similar in lean and obese animals, BWA1433 administration was found to lower the PTPase activities in the fractions obtained from obese but not from the lean rats. PTPases may be upregulated in muscles from obese rats due to activated adenosine receptors. Adenosine receptor blockade, by reducing PTPase activity, may thereby increase insulin signaling. PMID- 11404186 TI - S-myotrophin promotes the hypertrophy of myotube as insulin-like growth factor-I does. AB - We have reported previously that a novel muscle cell growth factor, having a structure of a peptide with sugar chains, was successfully purified from porcine skeletal muscle. It was named s-myotrophin. To determine the role of s-myotrophin in skeletal muscle growth, the effect of s-myotrophin on primary cultured chick skeletal muscle cells (composed almost totally of multinucleated myotubes) was investigated by comparing s-myotrophin with Insulin-like growth factor-I (IGF-I). Both s-myotrophin and IGF-I significantly increased creatine kinase activity of the cultures; both substances gave similar responses. Intracellar protein content was also increased by the addition of these factors. The content of myosin and actin in s-myotrophin treated culture in the differentiation medium was significantly higher than that of the control (unstimulated). The content of those proteins in IGF-I treated culture was also higher than that of control, but the differences were not statistically significant. Immunoblot analysis confirmed that the amounts of myosin and actin in the myocytes were greatly increased by s myotrophin stimulation and also by IGF-I stimulation. Morphological observations using an anti-desmin antibody staining procedure demonstrated that the size of both s-myotrophin and IGF-I treated myotubes was appreciably larger than that of control myotubes. These results suggest that s-myotrophin is a potent mediator of skeletal muscle cell hypertrophy thorough the accumulations of muscle structural proteins. PMID- 11404187 TI - Selection and timing of liver transplantation in primary biliary cirrhosis. PMID- 11404188 TI - False positive reaction in ELISA for IgM class anti-M2 antibody and its prevention. AB - Anti-M2 of anti-mitochondrial antibodies is recognized as the specific autoantibody detected in sera from patients with primary biliary cirrhosis (PBC). The IgG class and IgM class of this antibody can be separately measured using each ELISA. In the present study, false positive reactions were found in some sera from non-PBC patients such as acute hepatitis A, syphilis and rheumatoid arthritis using the IgM anti-M2 ELISA. They showed an increase of polyclonal IgM, and positivity for IgM anti-cardiolipin or rheumatoid factors, respectively. So, we developed a means to prevent these false positive reactions. First, dilutions of test sera at 1:1000-fold were carried out in addition to the original method at 1:100-fold. Secondly, some blocking reagents were added into the buffer system. By serum dilution, non-specific bindings disappeared in most samples other than showing an increase in polyclonal IgM. Moreover, the addition of suitable blocking reagents such as fetal bovine serum (FBS) and skimmed milk into the buffer system could prevent these non-specific bindings. From these findings, the procedure of optical serum dilution and the addition of suitable blocking reagents successfully prevented false positive reactions in this IgM anti-M2 ELISA. PMID- 11404189 TI - Influence of TT virus infection on the thrombocytopenia of patients with chronic liver disease. AB - TT virus (TTV) can replicate not only in the liver but also in hematopoietic cells. To investigate the possible influence of TTV infection on the hematological parameters of patients with chronic liver disease, serum samples from 581 patients with chronic, viral or non-viral liver disease were tested for TTV DNA by polymerase chain reaction (PCR). PCR was performed by two distinct methods using N22 primer pairs (N22 PCR), which detect primarily the four major TTV genotypes (1-4), and using genotype 1-specific primers (Genotype 1-PCR). Thirteen hematological parameters measured in routine laboratory tests were compared among the patients who were positive or negative for TTV infection. The platelet count was significantly lower in the N22 PCR-positive group than in the N22 PCR-negative group among the 189 patients with chronic hepatitis C (CH-C) and among the 40 patients with chronic hepatitis without serological markers of ongoing hepatitis B virus or hepatitis C virus infection (CH-NBNC) (P<0.0001, P<0.005, respectively). Similarly, among the patients with CH-C or CH-NBNC, the platelet count was significantly lower in the TTV genotype 1-positive group than in the TTV genotype 1-negative group (P<0.001, P<0.05, respectively). The reduced platelet count of the TTV DNA-positive group was observed in every fibrotic stage of chronic hepatitis (F0-F3). These results suggest that infection of TTV of certain genotypes including genotype 1 may worsen the thrombocytopenia of patients with CH-C or CH-NBNC, irrespective of the degree of hepatic fibrosis. PMID- 11404190 TI - Interferon alpha inhibits intrahepatic recurrence in hepatocellular carcinoma with chronic hepatitis C: a pilot study. AB - The aim of the present study is to evaluate whether interferon alpha (IFNalpha) therapy can inhibit intrahepatic recurrence after the curative treatment of small HCC with underlying chronic hepatitis C. Forty patients were enrolled in this study. They had solitary, small HCC90% of the area showed positive staining), moderately positive (5-90% of the area), weakly positive (<5% of the area), and negative (completely negative). All 19 (100%) well-differentiated, trabecular type HCC areas were strongly positive for Hep Par 1. Among 11 well differentiated, pseudoglandular type HCC areas, 2 (18%) were strongly positive, 5 (46%) were moderately positive, 2 (18%) were weakly positive, and 2 (18%) were negative. Among 36 moderately differentiated, trabecular type HCC areas, 6 (17%) were strongly positive, 17 (47%) were moderately positive, 9 (25%) were weakly positive, and 4 (11%) were negative. None of the four moderately differentiated, pseudoglandular type HCC areas were strongly positive, 3 (75%) were moderately positive, 0 was weakly positive, and 1 (25%) was negative. Among 25 poorly differentiated, compact or trabecular type HCC areas, 15 (60%) were weakly positive and 10 (40%) were negative. All 5 (100%) undifferentiated HCC areas were negative for Hep Par 1. Our results indicate that immunoreactivity to Hep Par 1 in HCC decreases with reduced differentiation of the tumor, suggesting that Hep Par 1 monoclonal antibody is useful as a marker for the diagnosis and differentiation of HCCs. PMID- 11404192 TI - Hepatic histopathologic range compared with virological studies of hepatitis viruses among autopsy cases in Tokyo. AB - Few reports exist comparing virological studies on hepatitis viruses with histopathological studies of autopsy cases other than those of liver clinics. Relations between hepatitis virus-related markers and hepatic histopathology were studied in 1044 autopsy cases (779 men and 265 women) at the Medical Examiner's Office, Tokyo. Heart blood was obtained at the autopsy, and the sera were submitted for virus-marker detection of HBV, HCV, and HGV/GBV-C. Hematoxylin and eosin-stained paraffin sections were used for histological assessment. Histopathologically, 463 cases were determined as so-called normal liver; among them 440 cases (95.0%) were negative for all hepatitis virus-related markers, but HBV-DNA was positive in 13 cases, three cases were positive for HCV-RNA (indicating a healthy carrier rate of HCV-RNA of 4.1%), and seven cases were positive for HGV/GBV-C RNA. The incidence of these three virus-related markers was low in cases with fatty liver and micronodular cirrhosis, but in cases with chronic hepatitis, macronodular cirrhosis and hepatocellular carcinoma, the incidence of HBV-DNA and HCV-RNA increased with advancing disease. A positive rate of anti-HBs or anti-HBc (HBV-Ab) or both was found between 30 and 50% in all histopathological groups, and no noticeable relations between the positive rate and microscopical changes were detected. The presence of HGV/GBV-C RNA seemed to be unrelated to hepatic inflammation or generalized inflammatory changes or both occurring together. The decadal age incidence of the virus-related markers and their incidence in various hepatic diseases are also reported. PMID- 11404193 TI - Hepatitis C virus core particle detected by immunoelectron microscopy and optical rotation technique. AB - Hepatitis C Virus (HCV) are 55-65 nm spherical particles, but the internal structure of the virion remains to be clarified. To clarify the morphology of HCV core particles, we performed an immune electron microscopy (IEM) using plasma samples from two blood donors with high HCV RNA titers and a detergent-treated anti-HCV core antibody-free plasma sample with high HCV RNA titer (1.5x10(8) copies/ml). Spherical particles, with 33-40 nm in diameter (an average diameter of 37 nm) were found in 1.22-1.25 g/ml fractions after sucrose density gradient centrifugation by conventional electron microscopy (EM). IEM using rabbit polyclonal antibody (RR8) specific to the putative HCV core protein and goat anti rabbit IgG colloidal gold particles revealed that these spherical particles specifically reacted with RR8. This finding indicates that the spherical particles are naked HCV core particles. Some of the HCV core particles had an icosahedron-like structure. Optical rotation technique showed that the HCV core particle exhibits six-fold symmetry and that the length of regular hexagon side is approximately 20 nm. These findings showed that HCV core particles are spherical particles of 33-40 nm in diameter and that HCV core particles may possess an icosahedron-like structure and a buoyant density of 1.22-1.25 g/ml. PMID- 11404194 TI - Retreatment with interferon-alpha at dosages or period increased by 1.3 times is effective for treatment for transient responders and non-responders in chronic hepatitis C patients. AB - The objectives of retreatment with interferon (IFN) in chronic hepatitis C (CH-C) patients are sustained response and a reduction in the risk of hepatocellular carcinoma (HCC). However, it is still unclear, as to which patients are candidates for retreatment with IFN. Eighteen transient responders (TRs) and 15 non-responders (NRs) to IFN therapy in CH-C received retreatment with IFNalpha. Of the 18 TRs, five showed sustained disappearance of hepatitis C virus, two showed sustained biochemical response, 10 continued as TR and one was a NR. Of the 15 NRs, six showed a TR while nine continued as NRs. Responsive cases, which included the virologically or biochemically sustained and transient responders, received either a dose of IFN 1.3 times greater or were treated for a period of 1.3 times longer in the retreatment than the original treatment. We submit that IFN treatment consisting of either a time period or a dosage 1.3 times those of the original IFN administration may be beneficial in the case of TR and NR in chronic hepatitis C patients. PMID- 11404195 TI - Combined treatment of HCV infection: is there a need for meta-analysis? AB - Background/Aims. The aim of this study was to estabilish the 'true' therapeutical gain of a combined therapy interferon - ribavirin versus interferon monotherapy in hepatitis C virus infection.Methods. A systematic review of published trials comparing combined treatment (interferon+ribavirin) vs interferon monotherapy, was performed on articles printed from 1991 to 1999.Results. The meta-analysis of retrieved trials showed that ribavirin significantly enhances the sustained response rate to interferon therapy in all types of patients. The results of almost all studies were homogeneous: combined therapy approximately doubles response rates compared to interferon monotherapy. At the end of the treatment, ribavirin increased the rate of viral clearance approximately 22% and the biochemical response of about 31%. The net gain was about 24% for both viral and biochemical response rate after 24 weeks of follow-up.Conclusion. Combined therapy showed a significantly higher efficacy in terms of sustained negativity of viral genome in all classes of patients (naives, relapsers, non-responders) becoming the therapy of choice. Nevertheless, more than about 50% of patients infected by hepatitis C virus show no sustained response to treatment. The whole weight of final data is beared by the major trials all with similar results. PMID- 11404196 TI - Photoprotection in adolescents. AB - Over the recent decades, there has been mounting medical evidence about the risks of ultraviolet light exposure and its potential for cutaneous malignancy. In the year 2000, an estimated one million Americans were diagnosed with skin cancer, which accounted for half of all new cancers. Worldwide, the incidence of cutaneous melanoma is increasing more rapidly than any other malignant neoplasm. Despite intensive educational efforts by the medical community, the public has not embraced adequate sun protection measures. This is especially true in adolescence, where peer acceptance and conflicting advertisements directly compete with the notion that pale is beautiful. There is strong evidence that preventive measures instituted early and encouraged throughout childhood and adolescence, can positively modify behavior. Towards this end, this article reviews the photobiology of ultraviolet light and its effects on the skin, as well as highlight practical information on sunscreens, adjunctive sun protection methods, sunless tanners, and tanning salons, which will help pediatricians educate adolescents and their families on proper photoprotective techniques. PMID- 11404197 TI - Pigmented lesions in adolescents. AB - Accurate diagnosis of congenital and acquired pigmented lesions accompanied by an understanding of their natural history and disease associations is critical for the appropriate management and counseling of adolescents, as well as timely referral to specialists when indicated. The recognition of atypical nevi and other melanoma risk factors in adolescents should lead to institution of preventive measures, such as routine skin examinations and counseling regarding sun protection. Because the incidence of melanoma is increasing in adolescents as well as adults, prompt identification of suspicious melanocytic lesions may lead to early diagnosis and effective treatment of melanoma. Numerous pigmented lesions can also herald the presence of a multisystem disorder; the recognition of syndromes associated with these lesions should result in appropriate evaluation and genetic counseling of affected individuals. This review distinguishes pigmented lesions that histologically represent a proliferation of melanocytes and that may therefore confer an increased risk for melanoma, from pigmented lesions due to increased melanization alone (i.e., increased melanin content) that are not associated with malignancy. PMID- 11404198 TI - Superficial fungal infections in adolescence. AB - Cutaneous fungal infections occur frequently in the adolescent population, and are second only to acne in generating skin-related adolescent anguish. Though superficial fungal infections of the skin are rarely life-threatening, they can cause significant worrisome cutaneous findings (e.g., white spots from pityriasis versicolor, distorted nails from onychomycosis). Such changes are particularly threatening to an adolescent who wants to be just like everybody else. This article focuses on several of the more common cutaneous fungal infections in adolescence, including pityrosporum (tinea) versicolor and dermatophyte infections of the feet, groin, skin, and nails. The prevalence of each disease is discussed, along with its clinical presentation, differential diagnosis, and methods of treatment. PMID- 11404199 TI - Management of warts and molluscum in adolescents. AB - Warts continue to be a therapeutic challenge, especially when they are numerous or widespread; in contrast, treatment of molluscum is comparatively more successful. A single, most effective treatment for either infection has not been defined. Conventional methods attempt to non-specifically destroy infected tissue. Most of these procedures are painful and require multiple treatments or compliance with daily application of a medication. The efficacy of destructive techniques has not been verified in placebo-controlled clinical trials and success rates reported in uncontrolled studies are difficult to interpret and often no better than that those achieved with a placebo. Alternative pharmacological approaches are aimed at stimulating immunologic or antiviral responses. Further study is needed to establish efficacy of these treatments. PMID- 11404200 TI - Bacterial skin infections in adolescents. AB - Bacterial skin infections are the single most common skin disorder for which pediatric patients seek care. This manuscript reviews the principles of skin infections and the most common and important bacterial skin infections in adolescents. PMID- 11404201 TI - Atopic, contact, and seborrheic dermatitis in adolescents. AB - Atopic dermatitis, contact dermatitis, and seborrheic dermatitis are chronic or recurrent inflammatory dermatoses that commonly affect adolescents. They may present a diagnostic and therapeutic challenge for physicians, and can be frustrating for patients. This article reviews the pathogenesis, diagnosis, and management of these diseases. PMID- 11404202 TI - Skin signs of sexually transmitted diseases in adolescents. AB - Sexually transmitted diseases (STDs) continue to be a significant and prevalent problem among adolescents. One in four sexually active teenagers are infected with one or more STDs. Certain STDs, including gonorrhea, chlamydia, and possibly human papillomavirus infections have the highest prevalence in this age group. The determinants of risk for STDs in adolescence encompass a range of behavioral, biological, and social factors. This review focuses on the skin signs of STDs in adolescents. Recognition of these findings is important since they may represent the most prominent, earliest, or only evidence of an STD. The recognition of the non-AIDs STDs has the added importance of identifying a population at high risk for HIV infection. Although recent data suggest encouraging trends in sexual risk behaviors among high school students in the United States, great challenges remain in the efforts to expand primary and secondary prevention strategies for STDs in adolescents. Expertise in the recognition and treatment of STDs in adolescents remains important for clinicians who care for adolescents. PMID- 11404203 TI - Sports dermatology. AB - Sports-related skin disorders are among the most common ailments affecting adolescent athletes and may significantly impact their performance and that of their team. These conditions may be due to infection, sun exposure, trauma, inflammation or chemical changes. Knowledge of the various disorders and their unique presentations are paramount to the caring for athletes. When making decisions regarding treatment, the clinician should consider the implications for both the individual athlete and the team. PMID- 11404204 TI - Hypersensitivity syndromes. AB - The hypersensitivity syndromes are a group of diseases in which the cutaneous vasculature is altered through immunologic or nonimmunologic mechanisms. These reactions range in severity from mild to life-threatening and can be triggered by drugs, infectious agents, foods, or environmental allergens. Although their causes may vary, the morphologic appearance of these entities may be similar making a clinical diagnosis a challenge. PMID- 11404205 TI - The impact of skin disease on the quality of life of adolescents. AB - Though relatively well-investigated in adults, the impact of skin disease on the health-related quality of life (HRQL) of adolescents has only recently begun to be systematically evaluated. The primary goal of this article is to critically review previous research that has examined the relationship between HRQL and three types of skin disease commonly experienced by adolescents, namely acne vulgaris, alopecia areata, and atopic dermatitis. A secondary goal is to provide strategies to adolescent medicine physicians that will enable them to more effectively manage the psychological components of their patients dermatological conditions. PMID- 11404206 TI - Managing adolescent acne. AB - Acne vulgaris, more commonly termed acne, is the most common skin disease treated by physicians. It affects an estimated 17 million people in the United States, including 85% or more of adolescents and young adults. Acne is a chronic condition that may last for years and cause emotional distress and permanent scarring. Although there is no cure, medications can control the disease and limit or prevent scar formation. PMID- 11404208 TI - Development of bimanual skill: the search for stable patterns of coordination. AB - In 2 experiments, dynamic systems theory predictions concerning intrinsic dynamics and variability of bimanual coordination were examined at different developmental stages. In Experiment 1, ten 4-, 6-, 7-, 8-, and 10-year-old children and adults performed unimanual dominant, unimanual nondominant, and bimanual continuous circle drawing. All tasks were performed at the participants' preferred rate, size, and mode of coordination. The 4-, 6-, and 7-year-old children produced larger circles with longer durations than those of the 8- and 10-year-olds and the adults. That finding demonstrates that younger children display different intrinsic dynamics than older children and adults. The 4-, 6-, and 7-year-old children also displayed more variability in bimanual coordination (more time in less stable patterns of coordination, higher standard deviation in relative phase) and produced more transitions between coordination patterns than the 8- and 10-year-olds and the adults. In Experiment 2, the same participants performed bimanual circles at increasing rates. Consistent with predictions of the HKB model (H. Haken, J. A. S. Kelso, & H. Bunz, 1985), the number of transitions decreased as speed increased. Some support was found for the notion that age-related variables of attention and rate contribute to the increased variability in young children's bimanual coordination. PMID- 11404209 TI - Effects of an auditory model on the learning of relative and absolute timing. AB - The effects of an auditory model on the learning of relative and absolute timing were examined. In 2 experiments, participants attempted to learn to produce a 1,000- or 1,600-ms sequence of 5 key presses with a specific relative-timing pattern. In each experiment, participants were, or were not, provided an auditory model that consisted of a series of tones that were temporally spaced according to the criterion relative-timing pattern. In Experiment 1, participants (n = 14) given the auditory template exhibited better relative- and absolute-timing performance than participants (n = 14) not given the auditory template. In Experiment 2, auditory and no-auditory template groups again were tested, but in that experiment each physical practice participant (n = 16) was paired during acquisition with an observer (n = 16). The observer was privy to all instructions as well as auditory and visual information that was provided the physical practice participant. The results replicated the results of Experiment 1: Relative-timing information was enhanced by the auditory template for both the physical and observation practice participants. Absolute timing was improved only when the auditory model was coupled with physical practice. Consistent with the proposal of D. M. Scully and K. M. Newell (1985), modeled timing information in physical and observational practice benefited the learning of the relative-timing features of the task, but physical practice was required to enhance absolute timing. PMID- 11404210 TI - Consistent and variable practice conditions: effects on relative and absolute timing. AB - The authors conducted the present experiments to resolve the discrepancy between studies in which relative-timing learning has been found to be enhanced by consistent practice conditions and contextual interference experiments in which relative-timing learning has been found to be enhanced more by random practice than by blocked practice. There were 40 participants in Experiment 1 and 48 in Experiment 2. The results of Experiment 1 extended previous findings: The learning of the relative-timing pattern was systematically enhanced by the degree to which the practice conditions promoted movement consistency (constant > blocked > serial > random). Experiment 2 provided evidence that the discrepancy between the relative-timing effects in the 2 groups of studies was a product of the way in which relative-timing goals and feedback were presented. When the feedback was presented as segment times, random practice resulted in generally more stable relative-timing patterns during acquisition than blocked practice did. Thus, in both experiments, the learning of the relative-timing pattern was enhanced by more stable relative-timing conditions during acquisition. Absolute timing learning, as indexed by the transfer tests, was enhanced by serial or random practice as compared with constant or blocked practice, and was relatively unaffected by feedback conditions directed at the relative-timing pattern. In terms of motor programming theory, those findings are taken as additional evidence for the disassociation of memories supporting generalized motor program (GMP) performance, as indexed by relative timing, and parameter performance, as indexed by absolute timing. PMID- 11404211 TI - Changes in limb stiffness under conditions of mental stress. AB - In 2 experiments, the effects of mental stress on limb stiffness were investigated. The relative contribution to arm stiffness of individual muscle activity, co-contraction, muscle reflexes, and postural adjustments were examined. In each experiment, participants (N = 24, Experiment 1; N = 16, Experiment 2) held their supinated hand under a tray that they were required to return to horizontal after it had been suddenly released. Electromyographic activity in the biceps and triceps muscles was recorded, as were elbow and wrist angles and tray displacement. In Experiment 1, mental arithmetic stress was shown to lead to decreased tray displacement (i.e., increased resistance) compared with displacements under the control, unstressed condition, as well as to increased elbow flexion before tray release. In Experiment 2, the increased resistance to perturbation caused by mental stress was found to be independent of initial elbow angle, but to vary as a function of the amount of upward force exerted before tray release. The authors conclude that stress-induced increases in limb stiffness result from changes in the initial position of the elbow, specified by its angle, together with the initial force exerted by participants to counteract the mechanical perturbations. PMID- 11404212 TI - Interplay of biomechanical constraints and kinematic strategies in selecting arm postures. AB - In this study, the authors examined the interplay between biomechanics and control strategies in the resolution of excess degrees of freedom at the joint level. Seven participants made aimed arm movements from 30 starting points and several starting postures to targets. Final arm postures for movements to a target exhibited substantial joint angle variation. Through regression modeling and by comparing observed final arm postures with biomechanically plausible postures, the authors identified 3 kinematic strategies: (a) Maintain deviations from the average angle at the starting point to the joint's final posture; (b) make torso rotations that are a fixed proportion of shoulder rotations; and (c) adopt a characteristic combination of 4 wrist-positioning approaches. The results demonstrated that kinematic strategies can account for substantial variance in final arm postures, if one takes into account 2 types of individual differences those that arise inevitably from biomechanical constraints and those that reflect choices in movement strategy. PMID- 11404213 TI - Development of lower extremity kinetics for balance control in infants and young children. AB - Developmental changes in the kinematics and kinetics underlying balance control were studied in 61 children, 9 months to 10 years of age. The children were classified according to developmental milestones as standers; new, intermediate, and advanced walkers; runners-jumpers; hoppers; gallopers; and skippers. The children experienced support-surface translations of varying size and speed. Children with greater locomotor experience withstood larger balance threats without collapsing or stepping. Analyses of scaled trials (perturbations normalized in size to foot length and center of gravity height) revealed that improvement in balance was not related to initial configuration parameters surrounding the task (degree of crouch or lean). Children with advanced locomotor skills had faster recovery times and relatively larger muscle torques than children with less experience. Relative torque-time histories of the more experienced children began to match the adult response to similar perturbations. With increased experience and changing muscle torque regulatory abilities, balance skills became more robust. PMID- 11404214 TI - Stability in force-production tasks. AB - Exerting a force on a mechanical system can induce mechanical instability. To overcome that instability, humans may take advantage of their upper limb mechanical impedance (e.g., hand stiffness). The authors investigated what stiffness is required to maintain static stability and how humans can achieve that stiffness in the context of the task of pushing on a pivoting stick. Results showed that the stiffness required is in the range of measured human upper limb stiffness. To avoid an ill-posed problem, one can better express the requirements for stability as a simple geometrical criterion related to the curvature of the potential energy field at the hand. A planar model of the upper limb revealed that individuals can use both hand rotational and translational stiffness to stabilize a stick. Although hand rotational stiffness does not participate in producing the axial force on the stick, it can significantly contribute to achieving a limb stiffness appropriate for maintaining static stability. Hand rotational stiffness can be important for the design of hand tools, because humans can increase it only by augmenting grip force, a biomechanical factor associated with cumulative trauma injuries of the upper extremities. PMID- 11404215 TI - Coordination of eye and leg movements during visually guided stepping. AB - In the present study, 2 related hypotheses were tested: first, that vision is used in a feedforward control mode during precision stepping onto visual targets and, second, that the oculomotor and locomotor control centers interact to produce coordinated eye and leg movements during that task. Participants' (N = 4) eye movements and step cycle transition events were monitored while they performed a task requiring precise foot placement at every step onto irregularly placed stepping stones under conditions in which the availability of visual information was either restricted or intermittently removed altogether. Accurate saccades, followed by accurate steps, to the next footfall target were almost always made even when the information had been invisible for as long as 500 ms. Despite delays in footlift caused by the temporary removal (and subsequent reinstatement) of visual information, the mean interval between the start of the eye movement and the start of the swing toward a target did not vary significantly (p >.05). In contrast, the mean interval between saccade onset away from a target and a foot landing on that target (stance onset) did vary significantly (p <.05) under the different experimental conditions. Those results support the stated hypotheses. PMID- 11404216 TI - Knowledge of results for motor learning: relationship between error estimation and knowledge of results frequency. AB - The authors of the present study investigated the apparent contradiction between early and more recent views of knowledge of results (KR), the idea that how one is engaged before receiving KR may not be independent of how one uses that KR. In a 2 ×: 2 factorial design, participants (N = 64) practiced a simple force production task and (a) were required, or not required, to estimate error about their previous response and (b) were provided KR either after every response (100%) or after every 5th response (20%) during acquisition. A no-KR retention test revealed an interaction between acquisition error estimation and KR frequencies. The group that received 100% KR and was required to error estimate during acquisition performed the best during retention. The 2 groups that received 20% KR performed less well. Finally, the group that received 100% KR and was not required to error estimate during acquisition performed the poorest during retention. One general interpretation of that pattern of results is that motor learning is an increasing function of the degree to which participants use KR to test response hypotheses (J. A. Adams, 1971; R. A. Schmidt, 1975). Practicing simple responses coupled with error estimation may embody response hypotheses that can be tested with KR, thus benefiting motor learning most under a 100% KR condition. Practicing simple responses without error estimation is less likely to embody response hypothesis, however, which may increase the probability that participants will use KR to guide upcoming responses, thus attenuating motor learning under a 100% KR condition. The authors conclude, therefore, that how one is engaged before receiving KR may not be independent of how one uses KR. PMID- 11404217 TI - Magnitude and variation of ratio of total body potassium to fat-free mass: a cellular level modeling study. AB - Potassium is an essential element of living organisms that is found almost exclusively in the intracellular fluid compartment. The assumed constant ratio of total body potassium (TBK) to fat-free mass (FFM) is a cornerstone of the TBK method of estimating total body fat. Although the TBK-to-FFM (TBK/FFM) ratio has been assumed constant, a large range of individual and group values is recognized. The purpose of the present study was to undertake a comprehensive analysis of biological factors that cause variation in the TBK/FFM ratio. A theoretical TBK/FFM model was developed on the cellular body composition level. This physiological model includes six factors that combine to produce the observed TBK/FFM ratio. The ratio magnitude and range, as well as the differences in the TBK/FFM ratio between men and women and variation with growth, were examined with the proposed model. The ratio of extracellular water to intracellular water (E/I) is the major factor leading to between-individual variation in the TBK/FFM ratio. The present study provides a conceptual framework for examining the separate TBK/FFM determinants and suggests important limitations of the TBK/FFM method used in estimating total body fat in humans and other mammals. PMID- 11404218 TI - Inhibition of insulin signaling and glycogen synthesis by phorbol dibutyrate in rat skeletal muscle. AB - Numerous studies have shown a correlation between changes in protein kinase C (PKC) distribution and/or activity and insulin resistance in skeletal muscle. To investigate which PKC isoforms might be involved and how they affect insulin action and signaling, studies were carried out in rat soleus muscle incubated with phorbol esters. Muscles preincubated for 1 h with 1 microM phorbol 12,13 dibutyrate (PDBu) showed an impaired ability of insulin to stimulate glucose incorporation into glycogen and a translocation of PKC-alpha, -betaI, -theta, and -epsilon, and probably -betaII, from the cytosol to membranes. Preincubation with 1 microM PDBu decreased activation of the insulin receptor tyrosine kinase by insulin and to an even greater extent the phosphorylation of Akt/protein kinase B and glycogen synthase kinase-3. However, it failed to diminish the activation of phosphatidylinositol 3'-kinase by insulin. Despite these changes in signaling, the stimulation by insulin of glucose transport (2-deoxyglucose uptake) and glucose incorporation into lipid and oxidation to CO2 was unaffected. The results indicate that preincubation of skeletal muscle with phorbol ester leads to a translocation of multiple conventional and novel PKC isoforms and to an impairment of several, but not all, events in the insulin-signaling cascade. They also demonstrate that these changes are associated with an inhibition of insulin stimulated glycogen synthesis but that, at the concentration of PDBu used here, glucose transport, its incorporation into lipid, and its oxidation to CO2 are unaffected. PMID- 11404219 TI - Increased IGFR activity and glucose transport in cultured skeletal muscle from insulin receptor null mice. AB - We have studied the role of the insulin receptor (IR) in metabolic and growth promoting effects of insulin on primary cultures of skeletal muscle derived from the limb muscle of IR null mice. Cultures of IR null skeletal muscle displayed normal morphology and spontaneous contractile activity. Expression of muscle differentiating proteins was slightly reduced in myoblasts and myotubes of the IR null skeletal muscle cells, whereas that of the Na+/K+ pump appeared to be unchanged. Insulin-like growth factor receptor (IGFR) expression was higher in myoblasts from IR knockout (IRKO) than from IR wild-type (IRWT) mice but was essentially unchanged in myotubes. Expression of the GLUT-1 and GLUT-4 transporters appeared to be higher in IRKO than in IRWT myoblasts and was significantly greater in myotubes from IRKO than from IRWT cultures. Consistent with GLUT expression, both basal and insulin or insulin-like growth factor I (IGF I)-stimulated glucose uptakes were higher in IR null skeletal myotubes than in wild-type skeletal myotubes. Interestingly, autophosphorylation of IGFR induced by insulin and IGF-I was markedly increased in IR null skeletal myotubes. These results indicate that, in the absence of IR, there is a compensatory increase in basal as well as in insulin- and IGF-I-induced glucose transport, the former being mediated via increased activation of the IGF-I receptor. PMID- 11404220 TI - Zinc stimulates the activity of the insulin- and nutrient-regulated protein kinase mTOR. AB - Recent studies indicate that zinc activates p70 S6 kinase (p70(S6k)) by a mechanism involving phosphatidylinositol 3-kinase (PI 3-kinase) and Akt (protein kinase B). Here it is shown that phenanthroline, a zinc and heavy metal chelator, inhibited both amino acid- and insulin-stimulated phosphorylation of p70(S6k). Both amino acid and insulin activations of p70(S6k) involve a rapamycin-sensitive step that involves the mammalian target of rapamycin (mTOR, also known as FRAP and RAFT). However, in contrast to insulin, amino acids activate p70(S6k) by an unknown PI 3-kinase- and Akt-independent mechanism. Thus the effects of chelator on amino acid activation of p70(S6k) were surprising. For this reason, we tested the hypothesis that zinc directly regulates mTOR activity, independently of PI 3 kinase activation. In support of this, basal and amino acid stimulation of p70(S6k) phosphorylation was increased by zinc addition to the incubation media. Furthermore, the protein kinase activities of mTOR immunoprecipitated from rat brain lysates were stimulated two- to fivefold by 10-300 microM Zn2+ in the presence of an excess of either Mn2+ or Mg2+, whereas incubation with 1,10 phenanthroline had no effect. These findings indicate that Zn2+ regulates, but is not absolutely required for, mTOR protein kinase activity. Zinc also stimulated a recombinant human form of mTOR. The stimulatory effects of Zn2+ were maximal at approximately 100 microM but decreased and became inhibitory at higher physiologically irrelevant concentrations. Micromolar concentrations of other divalent cations, Ca2+, Fe2+, and Mn2+, had no effect on the protein kinase activity of mTOR in the presence of excess Mg2+. Our results and the results of others suggest that zinc acts at multiple steps in amino acid- and insulin cell signaling pathways, including mTOR, and that the additive effects of Zn2+ on these steps may thereby promote insulin and nutritional signaling. PMID- 11404221 TI - Mathematical model of FSH-induced cAMP production in ovarian follicles. AB - During the terminal part of their development, ovarian follicles become totally dependent on gonadotropin supply to pursue their growth and maturation. Both gonadotropins, follicle-stimulating hormone (FSH) and luteining hormone (LH), operate mainly through stimulatory G protein-coupled receptors, their signal being transduced by the activation of the enzyme adenylyl cyclase and the production of second-messenger cAMP. In this paper, we develop a mathematical model of the dynamics of the coupling between FSH receptor stimulation and cAMP synthesis. This model takes the form of a set of nonlinear, ordinary differential equations that describe the changes in the different states of FSH receptors (free, bound, phosphorylated, and internalized), coupling efficiency (activated adenylyl cyclase), and cAMP response. Classical analysis shows that, in the case of constant FSH signal input, the system converges to a unique, stable equilibrium state, whose properties are here investigated. The system also appears to be robust to nonconstant input. Particular attention is given to the influence of biologically relevant parameters on cAMP dynamics. PMID- 11404222 TI - Regional physiological adaptation of the central nervous system deiodinases to iodine deficiency. AB - The goal of the present investigation was to analyze the types 2 (D2) and 3 (D3) iodothyronine deiodinases in various structures within the central nervous system (CNS) in response to iodine deficiency. After 5-6 wk of low-iodine diet (LID) or LID + 2 microg potassium iodide/ml (LID + KI; control), rats' brains were processed for in situ hybridization histochemistry for D2 and D3 mRNA or dissected, frozen in liquid nitrogen, and processed for D2 and D3 activities. LID did not affect weight gain or serum triiodothyronine, but plasma thyroxine (T4) was undetectable. In the LID + KI animals, D3 activities were highest in the cerebral cortex (CO) and hippocampus (HI), followed by the olfactory bulb and was lowest in cerebellum (CE). Iodine deficiency decreased D3 mRNA expression in all CNS regions, and these changes were accompanied by three- to eightfold decreases in D3 activity. In control animals, D2 activity in the medial basal hypothalamus (MBH) was similar to that in pituitary gland. Of the CNS D2-expressing regions analyzed, the two most responsive to iodine deficiency were the CO and HI, in which an approximately 20-fold increase in D2 activity occurred. Other regions, i.e., CE, lateral hypothalamus, MBH, and pituitary gland, showed smaller increases. The distribution of and changes in D2 mRNA were similar to those of D2 activity. Our results indicate that decreases in the expression of D3 and increases in D2 are an integral peripheral component of the physiological response of the CNS to iodine deficiency. PMID- 11404223 TI - Prevention of skeletal muscle insulin resistance by dietary cod protein in high fat-fed rats. AB - In the present study, we tested the hypothesis that fish protein may represent a key constituent of fish with glucoregulatory activity. Three groups of rats were fed a high-fat diet in which the protein source was casein, fish (cod) protein, or soy protein; these groups were compared with a group of chow-fed controls. High-fat feeding led to severe whole body and skeletal muscle insulin resistance in casein- or soy protein-fed rats, as assessed by the euglycemic clamp technique coupled with measurements of 2-deoxy-D-[(3)H]glucose uptake rates by individual tissues. However, feeding cod protein fully prevented the development of insulin resistance in high fat-fed rats. These animals exhibited higher rates of insulin mediated muscle glucose disposal that were comparable to those of chow-fed rats. The beneficial effects of cod protein occurred without any reductions in body weight gain, adipose tissue accretion, or expression of tumor necrosis factor alpha in fat and muscle. Moreover, L6 myocytes exposed to cod protein-derived amino acids showed greater rates of insulin-stimulated glucose uptake compared with cells incubated with casein- or soy protein-derived amino acids. These data demonstrate that feeding cod protein prevents obesity-induced muscle insulin resistance in high fat-fed obese rats at least in part through a direct action of amino acids on insulin-stimulated glucose uptake in skeletal muscle cells. PMID- 11404224 TI - Glucose uptake and metabolism by cultured human skeletal muscle cells: rate limiting steps. AB - To use primary cultures of human skeletal muscle cells to establish defects in glucose metabolism that underlie clinical insulin resistance, it is necessary to define the rate-determining steps in glucose metabolism and to improve the insulin response attained in previous studies. We modified experimental conditions to achieve an insulin effect on 3-O-methylglucose transport that was more than twofold over basal. Glucose phosphorylation by hexokinase limits glucose metabolism in these cells, because the apparent Michaelis-Menten constant of coupled glucose transport and phosphorylation is intermediate between that of transport and that of the hexokinase and because rates of 2-deoxyglucose uptake and phosphorylation are less than those of glucose. The latter reflects a preference of hexokinase for glucose over 2-deoxyglucose. Cellular NAD(P)H autofluorescence, measured using two-photon excitation microscopy, is both sensitive to insulin and indicative of additional distal control steps in glucose metabolism. Whereas the predominant effect of insulin in human skeletal muscle cells is to enhance glucose transport, phosphorylation, and steps beyond, it also determines the overall rate of glucose metabolism. PMID- 11404225 TI - Role of the human V1 vasopressin receptor COOH terminus in internalization and mitogenic signal transduction. AB - We studied the role played by the intracellular COOH-terminal region of the human arginine vasopressin (AVP) V1-vascular receptor (V1R) in ligand binding, trafficking, and mitogenic signal transduction in Chinese hamster ovary cells stably transfected with the human AVP receptor cDNA clones that we had isolated previously. Truncations, mutations, or chimeric alterations of the V1R COOH terminus did not alter ligand binding, but agonist-induced V1R internalization and recycling were reduced in the absence of the proximal region of the V(1)R COOH terminus. Coupling to phospholipase C was altered as a function of the COOH terminal length. Deletion of the proximal portion of the V1R COOH terminus or its replacement by the V2-renal receptor COOH terminus prevented AVP stimulation of DNA synthesis and progression through the cell cycle. Mutation of a kinase consensus motif in the proximal region of the V1R COOH terminus also abolished the mitogenic response. Thus the V1R cytoplasmic COOH terminus is not involved in ligand specificity but is instrumental in receptor trafficking and facilitates the interaction between the intracellular loops of the receptor, G protein, and phospholipase C. It is absolutely required for transmission of the mitogenic action of AVP, probably via a specific kinase phosphorylation site. PMID- 11404226 TI - Anesthesia rapidly suppresses insulin pulse mass but enhances the orderliness of insulin secretory process. AB - Induction of anesthesia is accompanied by modest hyperglycemia and a decreased plasma insulin concentration. Most insulin is secreted in discrete pulses occurring at approximately 6- to 8-min intervals. We sought to test the hypothesis that anesthesia inhibits insulin release by disrupting pulsatile insulin secretion in a canine model by use of direct portal vein sampling. We report that induction of anesthesia causes an abrupt decrease in the insulin secretion rate (1.1 +/- 0.2 vs. 0.7 +/- 0.1 pmol. kg(-1). min(-1), P < 0.05) by suppressing insulin pulse mass (630 +/- 121 vs. 270 +/- 31 pmol, P < 0.01). Anesthesia also elicited an approximately 30% higher increase in insulin pulse frequency (P < 0.01) and more orderly insulin concentration profiles (P < 0.01). These effects were evoked by either sodium thiamylal or nitrous oxide and isoflurane. In conclusion, anesthesia represses insulin secretion through the mechanism of a twofold blunting of pulse mass despite an increase in orderly pulse frequency. These data thus unveil independent amplitude and frequency controls of beta-cells' secretory activity in vivo. PMID- 11404227 TI - A mathematical model of compartmentalized neurotransmitter metabolism in the human brain. AB - After administration of enriched [1-13C]glucose, the rate of 13C label incorporation into glutamate C4, C3, and C2, glutamine C4, C3, and C2, and aspartate C2 and C3 was simultaneously measured in six normal subjects by 13C NMR at 4 Tesla in 45-ml volumes encompassing the visual cortex. The resulting eight time courses were simultaneously fitted to a mathematical model. The rate of (neuronal) tricarboxylic acid cycle flux (V(PDH)), 0.57 +/- 0.06 micromol. g(-1). min(-1), was comparable to the exchange rate between (mitochondrial) 2 oxoglutarate and (cytosolic) glutamate (Vx), 0.57 +/- 0.19 micromol. g(-1). min( 1)), which may reflect to a large extent malate-aspartate shuttle activity. At rest, oxidative glucose consumption [CMR(Glc(ox))] was 0.41 +/- 0.03 miccromol. g(-1). min(-1), and (glial) pyruvate carboxylation (VPC) was 0.09 +/- 0.02 micromol. g(-1). min(-1). The flux through glutamine synthetase (Vsyn) was 0.26 +/- 0.06 micromol. g(-1). min(-1). A fraction of Vsyn was attributed to be from (neuronal) glutamate, and the corresponding rate of apparent glutamatergic neurotransmission (VNT) was 0.17 +/- 0.05 micromol. g(-1). min(-1). The ratio [VNT/CMR(Glcox)] was 0.41 +/- 0.14 and thus clearly different from a 1:1 stoichiometry, consistent with a significant fraction (approximately 90%) of ATP generated in astrocytes being oxidative. The study underlines the importance of assumptions made in modeling 13C labeling data in brain. PMID- 11404228 TI - Determinants of insulin-stimulated glucose disposal in middle-aged, premenopausal women. AB - Controversy exists regarding the relative importance of adiposity, physical fitness, and physical activity in the regulation of insulin-stimulated glucose disposal. To address this issue, we measured insulin-stimulated glucose disposal [mg. kg fat-free mass (FFM)(-1). min(-1); oxidative and nonoxidative components] in 45 nondiabetic, nonobese, premenopausal women (mean +/- SD; 47 +/- 3 yr) by use of hyperinsulinemic euglycemic clamp (40 mU. m(-2). min(-1)) and [6,6 2H2]glucose dilution techniques. We also measured body composition, abdominal fat distribution, thigh muscle fat content, maximal oxygen consumption (VO2 max), and physical activity energy expenditure ((2)H(2)(18)O kinetics) as possible correlates of glucose disposal. VO2 max was the strongest correlate of glucose disposal (r = 0.63, P < 0.01), whereas whole body and abdominal adiposity showed modest associations (range of r values from -0.32 to -0.46, P < 0.05 to P < 0.01). A similar pattern of correlations was observed for nonoxidative glucose disposal. None of the variables measured correlated with oxidative glucose disposal. The relationship of VO2 max to glucose disposal persisted after statistical control for FFM, percent body fat, and intra-abdominal fat (r = 0.40, P < 0.01). In contrast, correlations of total and regional adiposity measures to insulin sensitivity were no longer significant after statistical adjustment for VO2 max. VO2 max was the only variable to enter stepwise regression models as a significant predictor of total and nonoxidative glucose disposal. Our results highlight the importance of VO2 max as a determinant of glucose disposal and suggest that it may be a stronger determinant of variation in glucose disposal than total and regional adiposity in nonobese, nondiabetic, premenopausal women. PMID- 11404229 TI - Stimulatory effects of leptin and muscle contraction on fatty acid metabolism are not additive. AB - Leptin has been shown to acutely stimulate fatty acid oxidation and triacylglycerol hydrolysis in skeletal muscle. These effects are similar to those induced by muscle contraction alone. Several studies have demonstrated that, during aerobic exercise, plasma leptin concentrations are well maintained; however, none has examined whether the stimulatory effects of leptin and contraction on muscle lipid metabolism are additive. This is the first study to examine the direct effect of leptin on lipid and carbohydrate (CHO) metabolism in isolated oxidative muscle over a range of contraction intensities. We examined the effect of leptin (10 microg/ml) on the synthesis and degradation of muscle lipid pools [phospholipid (PL), diacylglycerol (DG), triacylglycerol (TG)] and palmitate oxidation in isolated resting and contracting (2, 8, and 20 tetani/min) soleus muscles. At rest, leptin increased fatty acid oxidation (+ 40%, P < 0.05) and TG hydrolysis (+ 47%, P < 0.05), while blunting TG esterification (-20%, P < 0.05). Glucose oxidation was unaffected at rest in the presence of leptin. During tetanic contraction, fatty acid oxidation (+20-114%, P < 0.05) and TG esterification (+ 19-33%, P < 0.05) as well as net TG utilization (+ 23%, P < 0.05) were all significantly increased. However, leptin was without further effect on any of these parameters during contraction. Net utilization of intramuscular glycogen, as well as glucose oxidation, was unaffected during contraction by leptin. The findings of the present study indicate that leptin has an important influence on lipid metabolism in resting muscle, but not during contraction. PMID- 11404230 TI - Effect of the GABAA agonist gaboxadol on nocturnal sleep and hormone secretion in healthy elderly subjects. AB - Aging is associated with a dramatic decrease in sleep intensity and continuity. The selective GABA(A) receptor agonist gaboxadol has been shown to increase non REM sleep and the duration of the non-REM episodes in rats and sleep efficiency in young subjects and to enhance low-frequency activity in the electroencephalogram (EEG) within non-REM sleep in both rats and humans. In this double-blind, placebo-controlled study, we investigated the influence of an oral dose of 15 mg of gaboxadol on nocturnal sleep and hormone secretion (ACTH, cortisol, prolactin, growth hormone) in 10 healthy elderly subjects (6 women). Compared with placebo, gaboxadol did not affect endocrine activity but significantly reduced perceived sleep latency, elevated self-estimated total sleep time, and increased sleep efficiency by decreasing intermittent wakefulness and powerfully augmented low-frequency activity in the EEG within non-REM sleep. These findings indicate that gaboxadol is able to increase sleep consolidation and non-REM sleep intensity, without disrupting REM sleep, in elderly individuals and that these effects are not mediated by a modulation of hormone secretion. PMID- 11404231 TI - Differential regulation of IGFBP-2 and IGFBP-5 gene expression by vitamin A status in Japanese quail. AB - To investigate the involvement of the insulin-like growth factor (IGF) system in vitamin A (VA)-supported growth, we examined the effects of VA status on IGF binding protein (IGFBP)-2 and -5 gene expression in Japanese quail. VA deficiency caused a reduction in IGFBP-2 mRNA only in lung, without effect in other tissues. However, the expression of IGFBP-5 mRNA was more sensitive to the change of VA status. IGFBP-5 mRNA levels were significantly reduced by VA depletion in a tissue-specific manner, which preceded the decrease in body weight. A single injection of retinoic acid or retinol to VA-deficient quail did not affect the levels of IGFBP-2 mRNA, but it rapidly induced the expression of IGFBP-5 mRNAs in some tissues. These results are the first to show that gene expression of some IGFBPs in vivo are under the control of VA status and suggest a possible involvement of the IGF system in mediating the physiological actions of VA in the growth of Japanese quail. PMID- 11404232 TI - Increased expression of GPI-specific phospholipase D in mouse models of type 1 diabetes. AB - Glycosylphosphatidylinositol-specific phospholipase D (GPI-PLD) is a high-density lipoprotein-associated protein. However, the tissue source(s) for circulating GPI PLD and whether serum levels are regulated are unknown. Because the diabetic state alters lipoprotein metabolism, and liver and pancreatic islets are possible sources of GPI-PLD, we hypothesized that GPI-PLD levels would be altered in diabetes. GPI-PLD serum activity and liver mRNA were examined in two mouse models of type 1 diabetes, a nonobese diabetic (NOD) mouse model and low-dose streptozotocin-induced diabetes in CD-1 mice. With the onset of hyperglycemia (2- to 5-fold increase over nondiabetic levels), GPI-PLD serum activity and liver mRNA increased 2- to 4-fold in both models. Conversely, islet expression of GPI PLD was absent as determined by immunofluorescence. Insulin may regulate GPI-PLD expression, because insulin treatment of diabetic NOD mice corrected the hyperglycemia along with reducing serum GPI-PLD activity and liver mRNA. Our data demonstrate that serum GPI-PLD levels are altered in the diabetic state and are consistent with liver as a contributor to circulating GPI-PLD. PMID- 11404233 TI - Exendin-4 reduces fasting and postprandial glucose and decreases energy intake in healthy volunteers. AB - Exendin-4 is a long-acting potent agonist of the glucagon-like peptide 1 (GLP-1) receptor and may be useful in the treatment of type 2 diabetes and obesity. We examined the effects of an intravenous infusion of exendin-4 (0.05 pmol. kg(-1). min(-1)) compared with a control saline infusion in healthy volunteers. Exendin-4 reduced fasting plasma glucose levels and reduced the peak change of postprandial glucose from baseline (exendin-4, 1.5 +/- 0.3 vs. saline, 2.2 +/- 0.3 mmol/l, P < 0.05). Gastric emptying was delayed, as measured by the paracetamol absorption method. Volunteers consumed 19% fewer calories at a free-choice buffet lunch with exendin-4 (exendin-4, 867 +/- 79 vs. saline 1,075 +/- 93 kcal, P = 0.012), without reported side effects. Thus our results are in accord with the possibility that exendin-4 may be a potential treatment for type 2 diabetes, particularly for obese patients, because it acts to reduce plasma glucose at least partly by a delay in gastric emptying, as well as by reducing calorie intake. PMID- 11404234 TI - Interrelationship between signal transduction pathways and 1,25(OH)2D3 in UMR106 osteoblastic cells. AB - In this study, the interrelationship between signal transduction pathways and 1,25-dihydroxyvitamin D(3) [1,25(OH)2D3] action was examined in UMR106 osteoblastic cells. Treatment of these cells with 8-bromo-cAMP (1 mM) resulted in an upregulation of the vitamin D receptor (VDR) and an augmentation in the induction by 1,25(OH)2D3 of 25(OH)D3 24-hydroxylase [24(OH)ase] and osteopontin (OPN) mRNAs as well as gene transcription. Transfection with constructs containing the vitamin D response element devoid of other promoter regulatory elements did not alter the cAMP-mediated potentiation, suggesting that cAMP enhanced transcription is due, at least in part, to upregulation of VDR. Treatment with phorbol ester [12-O-tetradecanoyl-phorbol-13-acetate (TPA) 100 nM], an activator of protein kinase C, significantly enhanced 1,25(OH)2D3-induced OPN mRNA and transcription but had no effect on VDR or on 24(OH)ase mRNA or transcription. Studies using OPN promoter constructs indicate that TPA-enhanced OPN transcription is mediated by an effect on the OPN promoter separate from an effect on VDR. Thus interactions with signal transduction pathways can enhance 1,25(OH)2D3 induction of 24(OH)ase and OPN gene expression, and, through different mechanisms, changes in cellular phosphorylation may play a significant role in determining the effectiveness of 1,25(OH)2D3 on transcriptional control in cells expressing skeletal phenotypic properties. PMID- 11404235 TI - TPN-evoked dysfunction of islet lysosomal activity mediates impairment of glucose stimulated insulin release. AB - We examined the relation between nutrient-stimulated insulin secretion and the islet lysosome acid glucan-1,4-alpha-glucosidase system in rats undergoing total parenteral nutrition (TPN). During TPN treatment, serum glucose was normal, but free fatty acids, triglycerides, and cholesterol were elevated. Islets from TPN infused rats showed increased basal insulin release, a normal insulin response to cholinergic stimulation but a greatly impaired response when stimulated by glucose or alpha-ketoisocaproic acid. This impairment of glucose-stimulated insulin release was only slightly ameliorated by the carnitine palmitoyltransferase 1 inhibitor etomoxir. However, in parallel with the impaired insulin response to glucose, islets from TPN-infused animals displayed reduced activities of islet lysosomal enzymes including the acid glucan-1,4-alpha glucosidase, a putative key enzyme in nutrient-stimulated insulin release. By comparison, the same lysosomal enzymes were increased in liver tissue. Furthermore, in intact control islets, the pseudotetrasaccharide acarbose, a selective inhibitor of acid alpha-glucosidehydrolases, dose dependently suppressed islet acid glucan-1,4-alpha-glucosidase and acid alpha-glucosidase activities in parallel with an inhibitory action on glucose-stimulated insulin secretion. By contrast, when incubated with intact TPN islets, acarbose had no effect on either enzyme activity or glucose-induced insulin release. Moreover, when acarbose was added directly to TPN islet homogenates, the dose-response effect on the catalytic activity of the acid alpha-glucosidehydrolases was shifted to the right compared with control homogenates. We suggest that a general dysfunction of the islet lysosomal/vacuolar system and reduced catalytic activities of acid glucan-1,4-alpha-glucosidase and acid alpha-glucosidase may be important defects behind the impairment of the transduction mechanisms for nutrient-stimulated insulin release in islets from TPN-infused rats. PMID- 11404236 TI - Parathyroid cells express dihydropyridine-sensitive cation currents and L-type calcium channel subunits. AB - Parathyroid cells express Ca2+ -conducting currents that are activated by raising the extracellular Ca2+ concentration ([Ca2+]o). We investigated the sensitivity of these currents to dihydropyridines, the expression of voltage-dependent Ca(2+) channel (VDCC) subunits, and the effects of dihydropyridines on the intracellular free [Ca2+] ([Ca2+]i) and secretion in these cells. Dihydropyridine channel antagonists dose dependently suppressed Ca2+ -conducting currents, and agonists partially reversed the inhibitory effects of the antagonists in these cells. From a bovine parathyroid cDNA library, we isolated cDNA fragments encoding parts of an alpha(1S)- and a beta(3)-subunit of L-type Ca(2+) channels. The alpha(1S) subunit cDNA from the parathyroid represents an alternatively spliced variant lacking exon 29 of the corresponding gene. Northern blot analysis and immunocytochemistry confirmed the presence of transcripts and proteins for alpha(1)- and beta(3)-subunits in the parathyroid gland. The addition of dihydropyridines had no significant effects on high [Ca2+]o-induced changes in [Ca2+]i and parathyroid hormone (PTH) release. Thus our studies indicate that parathyroid cells express alternatively spliced L-type Ca2+ channel subunits, which do not modulate acute intracellular Ca2+ responses or changes in PTH release. PMID- 11404237 TI - Sympathovagal imbalance in hyperthyroidism. AB - We assessed sympathovagal balance in thyrotoxicosis. Fourteen patients with Graves' hyperthyroidism were studied before and after 7 days of treatment with propranolol (40 mg 3 times a day) and in the euthyroid state. Data were compared with those obtained in a group of age-, sex-, and weight-matched controls. Autonomic inputs to the heart were assessed by power spectral analysis of heart rate variability. Systemic exposure to sympathetic neurohormones was estimated on the basis of 24-h urinary catecholamine excretion. The spectral power in the high frequency domain was considerably reduced in hyperthyroid patients, indicating diminished vagal inputs to the heart. Increased heart rate and mid-frequency/high frequency power ratio in the presence of reduced total spectral power and increased urinary catecholamine excretion strongly suggest enhanced sympathetic inputs in thyrotoxicosis. All abnormal features of autonomic balance were completely restored to normal in the euthyroid state. beta-Adrenoceptor antagonism reduced heart rate in hyperthyroid patients but did not significantly affect heart rate variability or catecholamine excretion. This is in keeping with the concept of a joint disruption of sympathetic and vagal inputs to the heart underlying changes in heart rate variability. Thus thyrotoxicosis is characterized by profound sympathovagal imbalance, brought about by increased sympathetic activity in the presence of diminished vagal tone. PMID- 11404238 TI - Molecular basis of hypoxia-induced pulmonary vasoconstriction: role of voltage gated K+ channels. AB - The hypoxia-induced membrane depolarization and subsequent constriction of small resistance pulmonary arteries occurs, in part, via inhibition of vascular smooth muscle cell voltage-gated K+ (KV) channels open at the resting membrane potential. Pulmonary arterial smooth muscle cell KV channel expression, antibody based dissection of the pulmonary arterial smooth muscle cell K+ current, and the O2 sensitivity of cloned KV channels expressed in heterologous expression systems have all been examined to identify the molecular components of the pulmonary arterial O2-sensitive KV current. Likely components include Kv2.1/Kv9.3 and Kv1.2/Kv1.5 heteromeric channels and the Kv3.1b alpha-subunit. Although the mechanism of KV channel inhibition by hypoxia is unknown, it appears that KV alpha-subunits do not sense O2 directly. Rather, they are most likely inhibited through interaction with an unidentified O2 sensor and/or beta-subunit. This review summarizes the role of KV channels in hypoxic pulmonary vasoconstriction, the recent progress toward the identification of KV channel subunits involved in this response, and the possible mechanisms of KV channel regulation by hypoxia. PMID- 11404239 TI - CFTR trafficking and signaling in respiratory epithelium. PMID- 11404240 TI - Na+ transport in normal and CF human bronchial epithelial cells is inhibited by BAY 39-9437. AB - To test the hypothesis that Na+ transport in human bronchial epithelial (HBE) cells is regulated by a protease-mediated mechanism, we investigated the effects of BAY 39-9437, a recombinant Kunitz-type serine protease inhibitor, on amiloride sensitive short-circuit current of normal [non-cystic fibrosis (CF) cells] and CF HBE cells. Mucosal treatment of non-CF and CF HBE cells with BAY 39-9437 decreased the short-circuit current, with a half-life of approximately 45 min. At 90 min, BAY 39-9437 (470 nM) reduced Na+ transport by approximately 70%. The inhibitory effect of BAY 39-9437 was concentration dependent, with a half-maximal inhibitory concentration of approximately 25 nM. Na+ transport was restored to control levels, with a half-life of approximately 15 min, on washout of BAY 39 9437. In addition, trypsin (1 microM) rapidly reversed the inhibitory effect of BAY 39-9437. These data indicate that Na+ transport in HBE cells is activated by a BAY 39-9437-inhibitable, endogenously expressed serine protease. BAY 39-9437 inhibition of this serine protease maybe of therapeutic potential for the treatment of Na+ hyperabsorption in CF. PMID- 11404241 TI - Synthetic chloride channel restores glutathione secretion in cystic fibrosis airway epithelia. AB - Cystic fibrosis (CF), an inherited disease characterized by defective epithelial Cl- transport, damages lungs via chronic inflammation and oxidative stress. Glutathione, a major antioxidant in the epithelial lung lining fluid, is decreased in the apical fluid of CF airway epithelia due to reduced glutathione efflux (Gao L, Kim KJ, Yankaskas JR, and Forman HJ. Am J Physiol Lung Cell Mol Physiol 277: L113-L118, 1999). The present study examined the question of whether restoration of chloride transport would also restore glutathione secretion. We found that a Cl- channel-forming peptide (N-K4-M2GlyR) and a K+ channel activator (chlorzoxazone) increased Cl- secretion, measured as bumetanide-sensitive short circuit current, and glutathione efflux, measured by high-performance liquid chromatography, in a human CF airway epithelial cell line (CFT1). Addition of the peptide alone increased glutathione secretion (181 +/- 8% of the control value), whereas chlorzoxazone alone did not significantly affect glutathione efflux; however, chlorzoxazone potentiated the effect of the peptide on glutathione release (359 +/- 16% of the control value). These studies demonstrate that glutathione efflux is associated with apical chloride secretion, not with the CF transmembrane conductance regulator per se, and the defect of glutathione efflux in CF can be overcome pharmacologically. PMID- 11404242 TI - Antioxidant imbalance in the lungs of cystic fibrosis transmembrane conductance regulator protein mutant mice. AB - Recent studies suggest that the cystic fibrosis (CF) transmembrane conductance regulator (CFTR) protein modulates epithelial reduced glutathione (GSH) transport and when defective creates an antioxidant imbalance. To test whether the CFTR protein modulates lung antioxidant defenses in vivo, epithelial lining fluid (ELF) and lung tissue from CFTR knockout (CFTR-KO) and wild-type (WT) mice were compared for GSH content and the activities of glutathione reductase, glutathione peroxidase, and gamma-glutamyltransferase. In the CFTR-KO mice, the ELF concentration of GSH was decreased (51%) compared with that in WT mice. The concentration of GSH in the lung tissue of CFTR-KO mice, however, was not significantly different from that in WT mice. The activities of glutathione reductase and glutathione peroxidase in the lung tissue of CFTR-KO mice were significantly increased compared with those in WT mice (48 and 28%, respectively). Tissue lipid and DNA oxidation were evaluated by measurement of thiobarbituric acid-reactive substances and 8-hydroxy-2'-deoxyguanosine, respectively. The levels of thiobarbituric acid-reactive substances and 8-hydroxy 2'-deoxyguanosine in the lung tissue of CFTR-KO mice were significantly increased compared with those in WT mice. These data support our hypothesis that a mutation in the CFTR gene can affect the antioxidant defenses in the lung and may contribute to the exaggerated inflammatory response observed in CF. PMID- 11404243 TI - Chaperoning the maturation of the cystic fibrosis transmembrane conductance regulator. PMID- 11404244 TI - Sodium 4-phenylbutyrate downregulates HSC70 expression by facilitating mRNA degradation. AB - Intracellular trafficking of the DeltaF508 cystic fibrosis transmembrane conductance regulator (CFTR) is repaired by sodium 4-phenylbutyrate (4PBA) by an undetermined mechanism. 4PBA downregulates protein and mRNA expression of the heat shock cognate protein HSC70 (the constitutively expressed member of the 70 kDa heat shock protein family) by approximately 40-50% and decreases formation of a HSC70-DeltaF508 CFTR complex that may be important in the intracellular degradation of DeltaF508 CFTR. We examined the potential mechanisms by which 4PBA decreases HSC70 mRNA and protein expression. In IB3-1 cells, 1 mM 4PBA did not alter the activity of the Chinese hamster ovary HSC70 promoter or of a human HSC70 promoter fragment in luciferase reporter assays nor did it alter HSC70 mRNA synthesis in nuclear runoff assays. In contrast, preincubation with 4PBA increased the rate of HSC70 mRNA degradation by approximately 40%. The initial rate of 35S-HSC70 protein synthesis in 4PBA-treated IB3-1 cells was reduced by approximately 40%, consistent with the steady-state mRNA level, whereas its rate of degradation was unaltered by 4PBA. 4PBA also reduced the steady-state accumulation of (35)S-HSC70 by approximately 40%. These data suggest that 4PBA decreases the expression of HSC70 mRNA and protein by inducing cellular adaptations that result in the decreased stability of HSC70 mRNA. PMID- 11404245 TI - Partial restoration of defective chloride conductance in DeltaF508 CF mice by trimethylamine oxide. AB - This study was designed to test the in vivo efficacy of the chemical chaperone trimethylamine oxide (TMAO) in correcting the Cl- transport defect in a mouse model of cystic fibrosis (CF). Rectal potential difference (RPD) measurements were done in matched wild-type and DeltaF508 CF mice. Mice were treated by subcutaneous injections of TMAO. Wild-type mice demonstrated a forskolin stimulated, Cl--dependent hyperpolarization of -6.4 +/- 0.8 mV (n = 11), which was significantly increased to -13.1 +/- 1.4 mV after treatment with TMAO. DeltaF508 CF mice showed no significant responses to forskolin. Treatment with TMAO recovered a forskolin-activated RPD in DeltaF508 CF mice (-1.1 +/- 0.2 mV; n = 17) but not in CFTR null mice. The effects of TMAO were dose dependent, resulting in a slope of -0.4 +/- 0.1 mV x g(-1) x kg(-1) in DeltaF508 CF mice. The forskolin-stimulated RPD in TMAO-treated DeltaF508 CF mice was partially blocked by glibenclamide and further stimulated by apigenin. The total response to forskolin plus apigenin was -2.5 +/- 0.45 mV (n = 6 mice), corresponding to 39% of the response evoked by forskolin only in wild-type mice. PMID- 11404246 TI - Induction of HSP70 promotes DeltaF508 CFTR trafficking. AB - The DeltaF508 cystic fibrosis transmembrane conductance regulator (CFTR) is a temperature-sensitive trafficking mutant that is detected as an immature 160-kDa form (band B) in gel electrophoresis. The goal of this study was to test the hypothesis that HSP70, a member of the 70-kDa heat shock protein family, promotes DeltaF508 CFTR processing to the mature 180-kDa form (band C). Both pharmacological and genetic techniques were used to induce HSP70. IB3-1 cells were treated with sodium 4-phenylbutyrate (4PBA) to promote maturation of DeltaF508 CFTR to band C. A dose-dependent increase in band C and total cellular HSP70 was observed. Under these conditions, HSP70-CFTR complexes were increased and 70-kDa heat shock cognate protein-CFTR complexes were decreased. Increased DeltaF508 CFTR maturation was also seen after transfection with an HSP70 expression plasmid and exposure to glutamine, an inducer of HSP70. With immunofluorescence techniques, the increased appearance of CFTR band C correlated with CFTR distribution beyond the perinuclear regions. These data suggest that induction of HSP70 promotes DeltaF508 CFTR maturation and trafficking. PMID- 11404247 TI - Dysregulated NF-kappaB activation in cystic fibrosis: evidence for a primary inflammatory disorder. PMID- 11404248 TI - Activation of NF-kappaB in airway epithelial cells is dependent on CFTR trafficking and Cl- channel function. AB - Polymorphonuclear leukocyte-dominated airway inflammation is a major component of cystic fibrosis (CF) lung disease and may be associated with CF transmembrane conductance regulator (CFTR) dysfunction as well as infection. Mutant DeltaF508 CFTR is mistrafficked, accumulates in the endoplasmic reticulum (ER), and may cause "cell stress" and activation of nuclear factor (NF)-kappaB. G551D mutants also lack Cl- channel function, but CFTR is trafficked normally. We compared the effects of CFTR mutations on the endogenous activation of an NF-kappaB reporter construct. In transfected Chinese hamster ovary cells, the mistrafficked DeltaF508 allele caused a sevenfold activation of NF-kappaB compared with wild type CFTR or the G551D mutant (P < 0.001). NF-kappaB was also activated in 9/HTEo /pCep-R cells and in 16HBE/pcftr antisense cell lines, which lack CFTR Cl- channel function but do not accumulate mutant protein in the ER. This endogenous activation of NF-kappaB was associated with elevated interleukin-8 expression. Impaired CFTR Cl- channel activity as well as cell stress due to accumulation of mistrafficked CFTR in the ER contributes to the endogenous activation of NF kappaB in cells with the CFTR mutation. PMID- 11404249 TI - Dopamine regulates Na-K-ATPase in alveolar epithelial cells via MAPK-ERK dependent mechanisms. AB - Dopamine (DA) increases lung edema clearance by regulating vectorial Na+ transport and Na-K-ATPase in the pulmonary epithelium. We studied the role of the mitogen-activated protein kinase (MAPK)-extracellular signal-regulated kinase (ERK) pathway in the DA regulation of Na-K-ATPase in alveolar epithelial cells (AEC). Incubation of AEC with DA resulted in a rapid stimulation of ERK activity via dopaminergic type 2 receptors. Analysis of total RNA and protein showed a 1.5 fold increase in the Na-K-ATPase beta1-subunit mRNA levels and up to a fivefold increase in beta1-subunit protein abundance after DA stimulation, which was blocked by the MAPK kinase (MEK) inhibitors PD-98059 and U-0126. Also, the DA-ERK pathway stimulated the synthesis of a green fluorescent protein reporter gene driven by the beta1-subunit promoter, which indicates that DA regulates the Na-K ATPase beta1-subunit at the transcriptional level. The DA-mediated increase in beta1-subunit mRNA protein resulted in an increase in functional Na pumps in the basolateral membranes of alveolar type II cells. These results suggest that the MAPK-ERK pathway is an important mechanism in the regulation of Na-K-ATPase by DA in the alveolar epithelium. PMID- 11404250 TI - Involvement of the MAP kinase ERK2 in MUC1 mucin signaling. AB - MUC1 mucin is a receptor-like glycoprotein expressed abundantly in various cancer cell lines as well as in glandular secretory epithelial cells, including airway surface epithelial cells. The role of this cell surface mucin in the airway is not known. In an attempt to understand the signaling mechanism of MUC1 mucin, we established a stable cell line from COS-7 cells expressing a chimeric receptor consisting of the extracellular and transmembrane domains of CD8 and the cytoplasmic (CT) domain of MUC1 mucin (CD8/MUC1 cells). We previously observed that treatment of these cells with anti-CD8 antibody resulted in tyrosine phosphorylation of the CT domain of the chimera. Here we report that treatment of CD8/MUC1 cells with anti-CD8 resulted in activation of extracellular signal regulated kinase (ERK) 2 as assessed by immunoblotting, kinase assay, and immunocytochemistry. The activation of ERK2 was completely blocked either by a dominant negative Ras mutant or in the presence of a mitogen-activated protein kinase kinase (MEK) inhibitor. We conclude that tyrosine phosphorylation of the CT domain of MUC1 mucin leads to activation of a mitogen-activated protein kinase pathway through the Ras-MEK-ERK2 pathway. Combined with the existing data by others, it is suggested that one of the roles of MUC1 mucin may be regulation of cell growth and differentiation via a common signaling pathway, namely the Grb2 Sos-Ras-MEK-ERK2 pathway. PMID- 11404251 TI - IL-12 attenuates bleomycin-induced pulmonary fibrosis. AB - Interleukin (IL)-12 is a potent inducer of interferon (IFN)-gamma. We postulated that IL-12 would attenuate bleomycin-induced pulmonary fibrosis. To test this hypothesis, we administered IL-12 or murine serum albumin to bleomycin-treated mice by daily intraperitoneal injection until day 12. Mice treated with IL-12 demonstrated decreased hydroxyproline levels compared with control treated mice. Furthermore, administration of IL-12 led to a time-dependent increase in both lung and bronchoalveolar lavage fluid IFN-gamma. The antifibrotic effect of IL-12 could be attenuated with simultaneous administration of neutralizing anti-IFN gamma antibodies. These findings support the notion that IL-12 attenuates bleomycin-induced pulmonary fibrosis via modulation of IFN-gamma production. PMID- 11404252 TI - Sphingomyelin metabolites inhibit sphingomyelin synthase and CTP:phosphocholine cytidylyltransferase. AB - Tissue injury in inflammation involves the release of several cytokines that activate sphingomyelinases and generate ceramide. In the lung, the impaired metabolism of surfactant phosphatidylcholine (PC) accompanies this acute and chronic injury. These effects are long-lived and extend beyond the time frame over which tumor necrosis factor (TNF)-alpha and interleukin-1beta are elevated. In this paper, we demonstrate that in H441 lung cells these two processes, cytokine-induced metabolism of sphingomyelin and the inhibition of PC metabolism, are directly interrelated. First, metabolites of sphingomyelin hydrolysis themselves inhibit key enzymes necessary for restoring homeostasis between sphingomyelin and its metabolites. Ceramide stimulates sphingomyelinases as effectively as TNF-alpha, thereby amplifying the sphingomyelinase activation, and TNF-alpha, ceramide, and sphingosine all inhibit PC:ceramide phosphocholine transferase (sphingomyelin synthase), the enzyme that restores homeostasis between sphingomyelin and ceramide pools. Second, ceramide inhibits PC synthesis, probably because of its effects on CTP:phosphocholine cytidylyltransferase, the rate-limiting enzymatic step in de novo PC synthesis. The data presented here suggest that TNF-alpha may be an inhibitor of phospholipid metabolism in inflammatory tissue injury. These actions may be amplified because of the ability of metabolites of sphingomyelin to inhibit the pathways that should restore the normal ceramide-sphingomyelin homeostasis. PMID- 11404253 TI - CTP:phosphocholine cytidylyltransferase inhibition by ceramide via PKC-alpha, p38 MAPK, cPLA2, and 5-lipoxygenase. AB - In a companion paper (Vivekananda J, Smith D, and King RJ. Am J Physiol Lung Cell Mol Physiol 281: L98-L107, 2001), we demonstrated that tumor necrosis factor (TNF)-alpha inhibited the activity of CTP:phosphocholine cytidylyltransferase (CT), the rate-limiting enzyme in the de novo synthesis of phosphatidylcholine (PC), and that its actions were likely exerted through a metabolite of sphingomyelin. In this paper, we explore the signaling pathway employed by TNF alpha using C2 ceramide as a cell-penetrating sphingolipid representative of the metabolites induced by TNF-alpha. We found that in H441 cells, as reported in other cell types, cytosolic phospholipase A2 (cPLA2) is activated by TNF-alpha. We also observed that the inhibiting action of C2 ceramide on CT requires protein kinase C-alpha, p38 mitogen-activated protein kinase, and cPLA2. The actions of C2 ceramide on CT activity can be duplicated by adding 2 microM lysoPC to these cells. Furthermore, we found that the effects of C2 ceramide are dependent on 5 lipoxygenase but that cyclooxygenase II is unimportant. We hypothesize that CT activity is inhibited by the lysoPC generated as a consequence of the activation of cPLA2 by protein kinase C-alpha and p38 mitogen-activated protein kinase. The other product of the activation of cPLA2, arachidonic acid, is a substrate for the synthesis of leukotrienes, which raise intracellular Ca2+ levels and complete the activation of cPLA2. PMID- 11404254 TI - Bacterial induction of pleural mesothelial monolayer barrier dysfunction. AB - Pneumonia remains one of the most common infectious causes of mortality. Patients with pneumonia develop parapneumonic effusions with a high neutrophil count as well as high protein concentrations. We hypothesized that pulmonary parenchymal bacterial infection causes a permeability change in the pleural mesothelium by inducing the production of vascular endothelial growth factor (VEGF). Complicated parapneumonic pleural effusions (empyema) have a 19-fold higher VEGF level than pleural fluids secondary to congestive heart failure and a 4-fold higher level than pleural fluids secondary to uncomplicated parapneumonic effusions. We also analyzed the influence of live Staphylococcus aureus on mesothelial barrier function using a model of confluent mesothelial monolayers. There was a significant drop in electrical resistance across S. aureus-infected pleural mesothelial cell (PMC) monolayers. Recombinant VEGF also decreases PMC electrical resistance. Neutralizing antibodies to VEGF significantly inhibited the drop in PMC electrical resistance caused by S. aureus. S. aureus infection also caused a significant increase in protein leak across confluent mesothelial monolayers. Our results suggest that bacterial pathogens induce VEGF release in mesothelial cells and alter mesothelial permeability, leading to protein exudation in empyema. PMID- 11404255 TI - Effects of cyclopiazonic acid on cytosolic calcium in bovine airway smooth muscle cells. AB - In many cells, inhibition of sarcoplasmic reticulum (SR) Ca2+-ATPase activity induces a steady-state increase in cytosolic calcium concentration ([Ca2+]i) that is sustained by calcium influx. The goal was to characterize the response to inhibition of SR Ca2+-ATPase activity in bovine airway smooth muscle cells. Cells were dispersed from bovine trachealis and loaded with fura 2-AM (0.5 microM) for imaging of single cells. Cyclopiazonic acid (CPA; 5 microM) inhibited refilling of both caffeine- and carbachol-sensitive calcium stores. In the presence of extracellular calcium, CPA caused a transient increase in [Ca2+]i from 166 +/- 11 to 671 +/- 100 nM, and then [Ca2+]i decreased to a sustained level (CPA plateau; 236 +/- 19 nM) significantly above basal. The CPA plateau spontaneously declined toward basal levels after 10 min and was attenuated by discharging intracellular calcium stores. When CPA was applied during sustained stimulation with caffeine or carbachol, decreases in [Ca2+]i were observed. We concluded that the CPA plateau depended on the presence of SR calcium and that SR Ca2+-ATPase activity contributed to sustained increases in [Ca2+]i during stimulation with caffeine and, to a lesser extent, carbachol. PMID- 11404256 TI - Evaluation of basement membrane degradation during TNF-alpha-induced increase in epithelial permeability. AB - We evaluated whether tumor necrosis factor (TNF)-alpha induces an increase in permeability of an alveolar epithelial monolayer via gelatinase secretion and basement membrane degradation. Gelatinase secretion and epithelial permeability to radiolabeled albumin under unstimulated and TNF-alpha-stimulated conditions of an A549 human epithelial cell line were evaluated in vitro. TNF-alpha induced both upregulation of a 92-kDa gelatinolytic activity (pro form in cell supernatant and activated form in extracellular matrix) and an increase in the epithelial permeability coefficient compared with the unstimulated condition (control: 1.34 +/- 0.04 x 10(-6) cm/s; 1 microg/ml TNF-alpha: 1.47 +/- 0.05 x 10( 6) cm/s, P < 0.05). The permeability increase in the TNF-alpha-stimulated condition involved both paracellular permeability, with gap formation visualized by actin cytoskeleton staining, and basement membrane permeability, with an increase in the basement membrane permeability coefficient (determined after cell removal; control: 2.58 +/- 0.07 x 10(-6) cm/s; 1 microg/ml TNF-alpha: 2.82 +/- 0.02.10(-6) x cm/s, P < 0.05). Because addition of gelatinase inhibitors [tissue inhibitor of metalloproteinase (TIMP)-1 or BB-3103] to cell supernatants failed to inhibit the permeability increase, the gelatinase-inhibitor balance in the cellular microenvironment was further evaluated by cell culture on a radiolabeled collagen matrix. In the unstimulated condition, spontaneous collagenolytic activity inhibited by addition to the matrix of 1 microg/ml TIMP-1 or 10(-6) M BB 3103 was found. TNF-alpha failed to increase this collagenolytic activity because it was associated with dose-dependent upregulation of TIMP-1 secretion by alveolar epithelial cells. In conclusion, induction by TNF-alpha of upregulation of both the 92-kDa gelatinase and its inhibitor TIMP-1 results in maintenance of the gelatinase-inhibitor balance, indicating that basement membrane degradation does not mediate the TNF-alpha-induced increase in alveolar epithelial monolayer permeability. PMID- 11404257 TI - Uptake of a natural surfactant and increased delivery of small organic anions into type II pneumocytes. AB - The uptake of natural lung surfactant into differentiated type II cells may be used for the targeted delivery of other molecules. The fluorescent anion pyranine [hydroxypyren-1,3,6-trisulfonic acid, sodium salt (HPTS)] was incorporated into a bovine surfactant labeled with [3H]dipalmitoylphosphatidylcholine ([3H]DPPC). The uptake of [3H]DPPC and of HPTS increased with time of incubation and concentration, decreased with the size of the vesicles used, and was stimulated by 8-bromo-cAMP and partially inhibited by hypertonic sucrose. However, the amount of HPTS uptake was approximately 100 times smaller than that of [3H]DPPC. This large difference was due to a more rapid regurgitation of some of the HPTS from the cells but not to leakage from the surfactant before uptake. The acidification of the internalized surfactant increased linearly over 90 min to 7.13, and after 24 h, a pH of 6.83 was measured. In conclusion, after internalization of a double-labeled natural surfactant, the lipid moieties were accumulated in relation to the anions, which were targeted to a compartment not very acidic and in part rapidly expelled from the cells. PMID- 11404258 TI - Tyrosine kinase receptor activation inhibits NPR-C in lung arterial smooth muscle cells. AB - We have previously demonstrated that expression of the atrial natriuretic peptide (ANP) clearance receptor (NPR-C) is reduced selectively in the lung of rats and mice exposed to hypoxia but not in pulmonary arterial smooth muscle cells (PASMCs) cultured under hypoxic conditions. The current study tested the hypothesis that hypoxia-responsive growth factors, fibroblast growth factors (FGF 1 and FGF-2) and platelet-derived growth factor-BB (PDGF-BB), that activate tyrosine kinase receptors can reduce expression of NPR-C in PASMCs independent of environmental oxygen tension. Growth-arrested rat PASMCs were incubated under hypoxic conditions (1% O2) for 24 h; with FGF-1, FGF-2, or PDGF-BB (0.1-20 ng/ml for 1-24 h); or with ANG II (1-100 nM), endothelin-1 (ET-1, 0.1 microM), ANP (0.1 microM), sodium nitroprusside (SNP, 0.1 microM), or 8-bromo-cGMP (0.1 mM) for 24 h under normoxic conditions. Steady-state NPR-C mRNA levels were assessed by Northern blot analysis. FGF-1, FGF-2, and PDGF-BB induced dose- and time dependent reduction of NPR-C mRNA expression within 1 h at a threshold concentration of 1 ng/ml; hypoxia, ANG II, ET-1, ANP, SNP, or cGMP did not decrease NPR-C mRNA levels in PASMCs under the above conditions. Downregulation of NPR-C expression by FGF-1, FGF-2, and PDGF-BB was inhibited by the selective FGF-1 receptor tyrosine kinase inhibitor PD-166866 and mitogen-activated protein/extracellular signal-regulated kinase inhibitors U-0126 and PD-98059. These results indicate that activation of tyrosine kinase receptors by hypoxia responsive growth factors, but neither hypoxia per se nor activation of G protein coupled receptors, inhibits NPR-C gene expression in PASMCs. These results suggest that FGF-1, FGF-2, and PDGF-BB play a role in the signal transduction pathway linking hypoxia to altered NPR-C expression in lung. PMID- 11404259 TI - Bradykinin augments fibroblast-mediated contraction of released collagen gels. AB - Bradykinin is a multifunctional mediator of inflammation believed to have a role in asthma, a disorder associated with remodeling of extracellular connective tissue. Using contraction of collagen gels as an in vitro model of wound contraction, we assessed the effects of bradykinin tissue on remodeling. Human fetal lung fibroblasts were embedded in type I collagen gels and cultured for 5 days. After release, the floating gels were cultured in the presence of bradykinin. Bradykinin significantly stimulated contraction in a concentration- and time-dependent manner. Coincubation with phosphoramidon augmented the effect of 10(-9) and 10(-8) M bradykinin. A B2 receptor antagonist attenuated the effect of bradykinin, whereas a B1 receptor antagonist had no effect, suggesting that the effect is mediated by the B2 receptor. An inhibitor of intracellular Ca2+ mobilization abolished the response; addition of EGTA to the culture medium attenuated the contraction of control gels but did not modulate the response to bradykinin. In contrast, the phospholipase C inhibitor U-73122 and the protein kinase C inhibitors staurosporine and GF-109203X attenuated the responses. These data suggest that by augmenting the contractility of fibroblasts, bradykinin may have an important role in remodeling of extracellular matrix that may result in tissue dysfunction in chronic inflammatory diseases, such as asthma. PMID- 11404260 TI - Cellular response of antioxidant metalloproteins in Cu/Zn SOD transgenic mice exposed to hyperoxia. AB - Ceruloplasmin, metallothionein, and ferritin are metal-binding proteins with potential antioxidant activity. Despite evidence that they are upregulated in pulmonary tissue after oxidative stress, little is known regarding their influence on trace metal homeostasis. In this study, we have used copper- and zinc-containing superoxide dismutase (Cu/Zn SOD) transgenic-overexpressing and gene knockout mice and hyperoxia to investigate the effects of chronic and acute oxidative stress on the expression of these metalloproteins and to identify their influence on copper, zinc, and iron homeostasis. We found that the oxidative stress-mediated induction of ceruloplasmin and metallothionein in the lung had no effect on tissue levels of copper, iron, or zinc. However, Cu/Zn SOD expression had a marked influence on hepatic copper and iron as well as circulating copper homeostasis. These results suggest that ceruloplasmin and metallothionein may function as antioxidants independent of their role in trace metal homeostasis and that Cu/Zn SOD functions in copper homeostasis via mechanisms distinct from its superoxide scavenging properties. PMID- 11404261 TI - Chronic hypoxia-induced spontaneous and rhythmic contractions in the rat main pulmonary artery. AB - The effect of chronic hypoxia (CH; 1-4 wk) on the electromechanical properties of the rat main pulmonary artery (MPA) was investigated. MPA rings obtained from rats exposed for 14 days to hypobaric (50.5 kPa) CH exhibited spontaneous and rhythmic contractions (SRCs) that were never observed in control (normoxic) rats. SRCs were unaffected by tetrodotoxin, phentolamine, BQ-123 and BQ-788, N-nitro-L arginine methyl ester, or endothelium removal. CH depolarized smooth muscle cells from -58.8 +/- 9 to -38.6 +/- 5.4 mV and increased the resting cytosolic Ca2+ concentration from 67.3 +/- 11.9 to 112.5 +/- 16.4 nM. CH also induced spontaneous spikelike depolarizations. All of these effects were inhibited by external Ca2+ removal or nifedipine (1 microM). Moreover, depletion of intracellular Ca2+ stores with ryanodine (1-5 microM) or cyclopiazonic acid (3 microM) progressively attenuated SRCs. This study demonstrates that CH switches the MPA from a quiescent to a spontaneously active mechanical state. Finally, the fact that SRCs precede the development of right ventricle hypertrophy and disappear when this hypertrophy reaches a maximal value (after 3-4 wk of CH) suggests that SRCs may play a role in the adaptive process of the pulmonary circulation to CH. PMID- 11404262 TI - Effect of chronic hypoxia on agonist-induced tone and calcium signaling in rat pulmonary artery. AB - The effect of chronic hypoxia (CH) for 14 days on Ca2+ signaling and contraction induced by agonists in the rat main pulmonary artery (MPA) was investigated. In MPA myocytes obtained from control (normoxic) rats, endothelin (ET)-1, angiotensin II (ANG II), and ATP induced oscillations in intracellular Ca2+ concentration ([Ca2+]i) in 85-90% of cells, whereas they disappeared in myocytes from chronically hypoxic rats together with a decrease in the percentage of responding cells. However, both the amount of mobilized Ca2+ and the sources of Ca2+ implicated in the agonist-induced response were not changed. Analysis of the transient caffeine-induced [Ca2+]i response revealed that recovery of the resting [Ca2+]i value was delayed in myocytes from chronically hypoxic rats. The maximal contraction induced by ET-1 or ANG II in MPA rings from chronically hypoxic rats was decreased by 30% compared with control values. Moreover, the D-600- and thapsigargin-resistant component of contraction was decreased by 40% in chronically hypoxic rats. These data indicate that CH alters pulmonary arterial reactivity as a consequence of an effect on both Ca2+ signaling and Ca2+ sensitivity of the contractile apparatus. A Ca2+ reuptake mechanism appears as a CH-sensitive phenomenon that may account for the main effect of CH on Ca2+ signaling. PMID- 11404263 TI - Partial HIF-1alpha deficiency impairs pulmonary arterial myocyte electrophysiological responses to hypoxia. AB - Chronic hypoxia depolarizes and reduces K+ current in pulmonary arterial smooth muscle cells (PASMCs). Our laboratory previously demonstrated that hypoxia inducible factor-1 (HIF-1) contributed to the development of hypoxic pulmonary hypertension. In this study, electrophysiological parameters were measured in PASMCs isolated from intrapulmonary arteries of mice with one null allele at the Hif1a locus encoding HIF-1alpha [Hif1a(+/-)] and from their wild-type [Hif1a(+/+)] littermates after 3 wk in air or 10% O2. Hematocrit and right ventricular wall and left ventricle plus septum weights were measured. Capacitance, K+ current, and membrane potential were measured with whole cell patch clamp. Similar to our laboratory's previous results, hypoxia-induced right ventricular hypertrophy and polycythemia were blunted in Hif1a(+/-) mice. Hypoxia increased PASMC capacitance in Hif1a(+/+) mice but not in Hif1a(+/-) mice. Chronic hypoxia depolarized and reduced K+ current density in PASMCs from Hif1a(+/+) mice. In PASMCs from hypoxic Hif1a(+/-) mice, no reduction in K+ current density was observed, and depolarization was significantly blunted. Thus partial deficiency of HIF-1alpha is sufficient to impair hypoxia-induced depolarization, reduction of K+ current density, and PASMC hypertrophy. PMID- 11404264 TI - Carbon monoxide attenuates aeroallergen-induced inflammation in mice. AB - Carbon monoxide (CO) generated by catalysis of heme by heme oxygenase is increased in the exhaled air of asthmatic patients. Based on recent studies demonstrating that asthma is an inflammatory disease associated with increased oxidants and that CO confers cytoprotection in oxidant-induced lung injury and inflammation, we sought to better understand the functional role of CO in asthma by using an aeroallergen model. Mice were sensitized to ovalbumin, challenged with aerosolized ovalbumin, and maintained in either CO (250 parts/million) or room air for 48 h. The differential effects of CO on bronchoalveolar lavage (BAL) fluid cell types were observed, with a marked attenuation of BAL fluid eosinophils in the CO-treated animals at 24 and 48 h. A marked reduction of the proinflammatory cytokine interleukin-5 was observed in the CO-treated mice, with no significant changes for other proinflammatory cytokines. These differential effects of CO were also observed with leukotrienes (LTs) and prostaglandins in that CO significantly decreased BAL fluid PGE2, and LTB4 but exerted negligible effect on thromboxane B2 or LTC4/D4/E4. Our data suggest a putative immunoregulatory role for CO in aeroallergen-induced inflammation in mice. PMID- 11404265 TI - Modulation of alpha-ENaC and alpha1-Na+-K+-ATPase by cAMP and dexamethasone in alveolar epithelial cells. AB - cAMP and dexamethasone are known to modulate Na+ transport in epithelial cells. We investigated whether dibutyryl cAMP (DBcAMP) and dexamethasone modulate the mRNA expression of two key elements of the Na+ transport system in isolated rat alveolar epithelial cells: alpha-, beta-, and gamma-subunits of the epithelial Na+ channel (ENaC) and the alpha1- and beta1-subunits of Na+-K+-ATPase. The cells were treated for up to 48 h with DBcAMP or dexamethasone to assess their long term impact on the steady-state level of ENaC and Na+-K+-ATPase mRNA. DBcAMP induced a twofold transient increase of alpha-ENaC and alpha1-Na+-K+-ATPase mRNA that peaked after 8 h of treatment. It also upregulated beta- and gamma-ENaC mRNA but not beta1-Na+-K+-ATPase mRNA. Dexamethasone augmented alpha-ENaC mRNA expression 4.4-fold in cells treated for 24 h and also upregulated beta- and gamma-ENaC mRNA. There was a 1.6-fold increase at 8 h of beta1-Na+-K+-ATPase mRNA but no significant modulation of alpha1-Na+-K+-ATPase mRNA expression. Because DBcAMP and dexamethasone did not increase the stability of alpha-ENaC mRNA, we cloned 3.2 kb of the 5' sequences flanking the mouse alpha-ENaC gene to study the impact of DBcAMP and dexamethasone on alpha-ENaC promoter activity. The promoter was able to drive basal expression of the chloramphenicol acetyltransferase (CAT) reporter gene in A549 cells. Dexamethasone increased the activity of the promoter by a factor of 5.9. To complete the study, the physiological effects of DBcAMP and dexamethasone were investigated by measuring transepithelial current in treated and control cells. DBcAMP and dexamethasone modulated transepithelial current with a time course reminiscent of the profile observed for alpha-ENaC mRNA expression. DBcAMP had a greater impact on transepithelial current (2.5-fold increase at 8 h) than dexamethasone (1.8-fold increase at 24 h). These results suggest that modulation of alpha-ENaC and Na+-K+-ATPase gene expression is one of the mechanisms that regulates Na+ transport in alveolar epithelial cells. PMID- 11404266 TI - Surfactant protein interactions with neutral and acidic phospholipid films. AB - The captive bubble tensiometer was employed to study interactions of phospholipid (PL) mixtures of dipalmitoylphosphatidylcholine (DPPC) and 1-palmitoyl-2-oleoyl sn-glycero-3-phosphocholine (POPC) or 1-palmitoyl-2-oleoyl-sn-glycero-3-[phospho rac-(1-glycerol)] (POPG) at 50 microg/ml with physiological levels of the surfactant protein (SP) A SP-B, and SP-C alone and in combination at 37 degrees C. All surfactant proteins enhanced lipid adsorption to equilibrium surface tension (gamma), with SP-C being most effective. Kinetics were consistent with the presence of two adsorption phases. Under the conditions employed, SP-A did not affect the rate of film formation in the presence of SP-B or SP-C. Little difference in gamma(min) was observed between the acidic POPG and the neutral POPC systems with SP-B or SP-C with and without SP-A. However, gamma(max) was lower with the acidic POPG system during dynamic, but not during quasi-static, cycling. Considerably lower compression ratios were required to generate low gamma(min) values with SP-B than SP-C. DPPC-POPG-SP-B was superior to the neutral POPC-SP-B system. Although SP-A had little effect on film formation with SP-B, surface activity during compression was enhanced with both PL systems. In the presence of SP-C, lower compression ratios were required with the acidic system, and with this mixture, SP-A addition adversely affected surface activity. The results suggest specific interactions between SP-B and phosphatidylglycerol, and between SP-B and SP-A. These observations are consistent with the presence of a surface-associated surfactant reservoir which is involved in generating low gamma during film compression and lipid respreading during film expansion. PMID- 11404267 TI - Role of zinc in pulmonary endothelial cell response to oxidative stress. AB - Although zinc is a well-known inhibitor of apoptosis, it may contribute to oxidative stress-induced necrosis. We noted that N,N,N',N'- tetrakis(2 pyridylmethyl)ethylenediamine (TPEN; >10 microM), a zinc chelator, quenched fluorescence of the zinc-specific fluorophore Zinquin and resulted in an increase in spontaneous apoptosis in cultured sheep pulmonary artery endothelial cells (SPAECs). Addition of exogenous zinc (in the presence of pyrithione, a zinc ionophore) to the medium of SPAECs caused an increase in Zinquin fluorescence and was associated with a concentration-dependent increase in necrotic cell death. Exposure of SPAECs to TPEN (10 microM) resulted in enhanced apoptosis after lipopolysaccharide or complete inhibition of t-butyl hydroperoxide (tBH)-induced necrosis. We further investigated the role of two zinc-dependent enzymes, poly(ADP-ribose) polymerase (PARP) and protein kinase (PK) C, in tBH toxicity. tBH toxicity was only affected by the PARP inhibitors 4-amino-1,8-naphthalimide or 3-aminobenzamide over a narrow range, whereas the PKC inhibitors bisindolylmaleimide and staurosporine significantly reduced tBH toxicity. tBH caused translocation of PKC to the plasma membrane of SPAECs that was partially inhibited by TPEN. Thus pulmonary endothelial cell zinc inhibits spontaneous and lipopolysaccharide-dependent apoptosis but contributes to tBH-induced necrosis, in part, via a PKC-dependent pathway. PMID- 11404268 TI - Novel mechanisms in murine nitrofen-induced pulmonary hypoplasia: FGF-10 rescue in culture. AB - We evaluated the role of the key pulmonary morphogenetic gene fibroblast growth factor-10 (Fgf10) in murine nitrofen-induced primary lung hypoplasia, which is evident before the time of diaphragm closure. In situ hybridization and competitive RT-PCR revealed a profound disturbance in the temporospatial pattern as well as a 10-fold decrease in mRNA expression level of Fgf10 but not of the inducible inhibitor murine Sprouty2 (mSpry2) after nitrofen treatment. Exogenous FGF-10 increased branching not only of control lungs [13% (right) and 27% (left); P < 0.01] but also of nitrofen-exposed lungs [23% (right) and 77% (left); P < 0.01]. Expression of mSpry2 increased 10-fold with FGF-10 in both nitrofen treated and control lungs, indicating intact downstream FGF signaling mechanisms after nitrofen treatment. We conclude that nitrofen inhibits Fgf10 expression, which is essential for lung growth and branching. Exogenous FGF-10 not only stimulates FGF signaling, marked by increased mSpry2 expression, in both nitrofen treated and control lungs but also substantially rescues nitrofen-induced lung hypoplasia in culture. PMID- 11404269 TI - Regulation of murine airway responsiveness by endothelial nitric oxide synthase. AB - Nitric oxide (NO) is a potent vasodilator, but it can also modulate contractile responses of the airway smooth muscle. Whether or not endothelial (e) NO synthase (NOS) contributes to the regulation of bronchial tone is unknown at present. Experiments were designed to investigate the isoforms of NOS that are expressed in murine airways and to determine whether or not the endogenous release of NO modulates bronchial tone in wild-type mice and in mice with targeted deletion of eNOS [eNOS(-/-)]. The presence of neuronal NOS (nNOS), inducible NOS (iNOS), and eNOS in murine trachea and lung parenchyma was assessed by RT-PCR, immunoblotting, and immunohistochemistry. Airway resistance was measured in conscious unrestrained mice by means of a whole body plethysmography chamber. The three isoforms of NOS were constitutively present in lungs of wild-type mice, whereas only iNOS and nNOS were present in eNOS(-/-) mice. Labeling of nNOS was localized in submucosal airway nerves but was not consistently detected, and iNOS immunoreactivity was observed in tracheal and bronchiolar epithelial cells, whereas eNOS was expressed in endothelial cells. In wild-type mice, treatment with N-nitro-L-arginine methyl ester, but not with aminoguanidine, potentiated the increase in airway resistance produced by inhalation of methacholine. eNOS(-/ ) mice were hyperresponsive to inhaled methacholine and markedly less sensitive to N-nitro-L-arginine methyl ester. These results demonstrate that the three NOS isoforms are expressed constitutively in murine lung and that NO derived from eNOS plays a physiological role in controlling bronchial airway reactivity. PMID- 11404270 TI - Enhanced clearance of surfactant protein D during LPS-induced acute inflammation in rat lung. AB - Pulmonary surfactant participates in the regulation of alveolar compliance and lung host defense. Surfactant homeostasis is regulated through a combination of synthesis, secretion, clearance, recycling, and degradation of surfactant components. The extracellular pool size of surfactant protein (SP) D fluctuates significantly during acute inflammation. We hypothesized that changes in SP-D levels are due, in part, to altered clearance of SP-D. Clearance pathways in rats were assessed with fluorescently labeled SP-D that was instilled into control lungs or lungs that had been treated with lipopolysaccharide (LPS) 16 h earlier. SP-D clearance from lavage into lung tissue was time dependent from 5 min to 1 h and 1.7-fold greater in LPS-treated lungs than in control lungs. Analysis of cells isolated by enzymatic digestion of lung tissue revealed differences in the SP-D-positive cell population between groups. LPS-treated lungs had 28.1-fold more SP-D-positive tissue-associated neutrophils and 193.6-fold greater SP-D association with those neutrophils compared with control lungs. These data suggest that clearance of SP-D into lung tissue is increased during inflammation and that tissue-associated neutrophils significantly contribute to this process. PMID- 11404271 TI - Nitric oxide modulates capillary formation at the endothelial cell-tumor cell interface. AB - Nitric oxide synthase expression has been documented in lung tumors, but a potential role for nitric oxide (NO) in induction of capillary formation remains to be elucidated. The purpose of this report was to characterize the direct effects of NO at the level of the tumor-endothelium interface with respect to angiogenesis. A Transwell two-compartment culture system, human endothelial cells (EC), and two human non-small cell lung cancer (CA) lines that constitutively produce NO were used to simulate the EC-tumor cell interface. Both histological types of lung CA, squamous and adenocarcinoma, induced baseline capillary formation by EC within 3 days. This process was inhibited by NO in the microenvironment because decreasing NO production with 100 microM aminoguanidine (AG) significantly increased capillary formation, whereas coincubation with 100 microM AG plus 400 microM L-arginine returned angiogenesis to baseline values. We demonstrate further that NO may exert its inhibitory effects by influencing matrix metalloproteinase expression/activity and tyrosine phosphorylation of proteins in the sprouting tips of nascent capillaries. PMID- 11404272 TI - Physiological levels and action of dehydroepiandrosterone in Yucatan miniature swine. AB - The biological role of dehydroepiandrosterone (DHEA) and its less active sulphated conjugate DHEAS was investigated in two experiments using Yucatan miniature swine. In experiment 1, plasma levels of both DHEA(S) among males were greater than female pigs that ranged in age from 0.3 to 84 mo old (P < 0.0001). In males, DHEA(S) were related inversely to serum triglycerides; DHEA was positively related to triglycerides in females (P < 0.01). In experiment 2, four 2-yr old male pigs, used as their own control, showed a 5% decrease in body weight, 11% increase in energy expenditure, 88% increase in lipid, and 100% decrease in glucose utilization (P < 0.0001) in response to DHEA vs. placebo treatments when adjusted for body weight. Plasma DHEA(S) were not different between treatment conditions. Glucose tolerance and plasma insulin levels were not different from controls. In vivo response to norepinephrine indicated beta adrenergic sensitivity was altered by DHEA. Present findings suggest DHEA and/or its hormone products are important in modulating energy expenditure and lipid utilization for energy in male animals. The role of DHEA in energy metabolism and the difference between sexes warrant further investigation. PMID- 11404273 TI - Ouabain-insensitive acidification by dopamine in renal OK cells: primary control of the Na(+)/H(+) exchanger. AB - The present study was aimed at evaluating the role of D(1)- and D(2)-like receptors and investigating whether inhibition of Na(+) transepithelial flux by dopamine is primarily dependent on inhibition of the apical Na(+)/H(+) exchanger, inhibition of the basolateral Na(+)-K(+)-ATPase, or both. The data presented here show that opossum kidney cells are endowed with D(1)- and D(2)-like receptors, the activation of the former, but not the latter, accompanied by stimulation of adenylyl cyclase (EC(50) = 220 +/- 2 nM), marked intracellular acidification (IC(50) = 58 +/- 2 nM), and attenuation of amphotericin B-induced decreases in short-circuit current (28.6 +/- 4.5% reduction) without affecting intracellular pH recovery after CO(2) removal. These results agree with the view that dopamine, through the activation of D(1)- but not D(2)-like receptors, inhibits both the Na(+)/H(+) exchanger (0.001933 +/- 0.000121 vs. 0.000887 +/- 0.000073 pH unit/s) and Na(+)-K(+)-ATPase without interfering with the Na(+)-independent HCO transporter. It is concluded that dopamine, through the action of D(1)-like receptors, inhibits both the Na(+)/H(+) exchanger and Na(+)-K(+)-ATPase, but its marked acidifying effects result from inhibition of the Na(+)/H(+) exchanger only, without interfering with the Na(+)-independent HCO transporter and Na(+) K(+)-ATPase. PMID- 11404274 TI - Lactate-transport activity in RBCs of trained and untrained individuals from four racing species. AB - In red blood cells (RBC) of horses, both lactate-transport activity and lactate accumulation during races vary interindividually. To study whether similar variation in lactate transport is apparent also in RBCs of other racing species, blood samples were collected from 21 reindeer, 40 horses, 31 humans, and 38 dogs. Total lactate-transport activity was measured at 10 and 30 mM concentrations, and the roles of the monocarboxylate-transporter (MCT) and the inorganic anion exchange transporter (band-3 protein) were studied with inhibitors. In the reindeer and in one-third of the horses, lactate transport was low and mediated mainly by band-3 protein and nonionic diffusion. In the humans, dogs, and the remaining two-thirds of the horses, lactate transport was high and MCT was the main transporter. No correlation existed between MCT activity and the athleticism of the species. In the horses and humans, training had no effect on lactate transport, but in the reindeer and sled dogs, training increased total lactate transport. These results show that among the racing species studied, only in horses was the distribution of lactate-transport activity bimodal, and the possible connection between RBC lactate and performance capacity, especially in this species, warrants further studies. PMID- 11404275 TI - Phenanthraquinone inhibits eNOS activity and suppresses vasorelaxation. AB - Diesel exhaust particles cause an impairment of endothelium-dependent vasorelaxation and are associated with cardiopulmonary-related diseases and mortality, but the mechanistic details are poorly understood. Since we reported previously that phenanthraquinone, an environmental chemical contained in diesel exhaust particles, suppresses neuronal nitric oxide synthase (nNOS) activity by shunting electrons away from the normal catalytic pathway, it was hypothesized that phenanthraquinone inhibits endothelial NOS (eNOS) activity and affects vascular tone. Therefore, the effects of phenanthraquinone on eNOS activity, endothelium-dependent relaxation, and blood pressure were examined in the present study. Phenanthraquinone inhibited NO formation evaluated by citrulline formed by total membrane fraction of bovine aortic endothelial cells with an IC(50) value of 0.6 microM. A kinetic study revealed that phenanthraquinone is a competitive inhibitor with respect to NADPH and a noncompetitive inhibitor with respect to L arginine. Endothelium-dependent relaxation of rat aortic rings by ACh was significantly inhibited by phenanthraquinone (5 microM), whereas the endothelium independent relaxation by nitroglycerin was not. Furthermore, an intraperitoneal injection of phenanthraquinone (0.36 mmol/kg) to rats resulted in an elevation of blood pressure (1.4-fold, P < 0.01); under this condition, plasma levels of stable NO metabolites, nitrite/nitrate, in phenanthraquinone-treated rats was reduced to 68% of control levels. The present findings suggest that phenanthraquinone has a potent inhibitory action on eNOS activity via a similar mechanism reported for nNOS, thereby causing the suppression of NO-mediated vasorelaxation and elevation of blood pressure. PMID- 11404276 TI - Hemodynamic response pattern predicts susceptibility to stress-induced elevation in arterial pressure in the rat. AB - Cocaine or air jet stress evokes pressor responses due to either a large increase in systemic vascular resistance (vascular responders) or small increases in both cardiac output and vascular resistance (mixed responders) in conscious rats. Repeated cocaine administration results in elevated arterial pressure in vascular responders but not in mixed responders. The present study examined the hypothesis that the pattern of cardiovascular responses to an unconditioned stimulus (UCS; air jet) is related to responses to a conditioned stimulus (CS; tone followed by brief foot shock) in individual rats. Our data demonstrate that presentation of the UCS produced variable cardiac output responses that correlated with responses to the CS (n = 60). We also determined whether individual cardiovascular response patterns to acute stress correlated with predisposition to a sustained stress induced elevation in arterial pressure. Rats were exposed to three different stressors presented one per day successively for 4 wk and during a poststress period of 3 wk while arterial pressure was recorded periodically. Mean arterial pressure was elevated in all rats during chronic stress but, during the poststress period, remained at significantly higher levels in vascular responders but not mixed responders. Therefore, we conclude that acute behavioral stress to a conditioned stimulus elicits variable hemodynamic responses that predict the predisposition to a sustained stress-induced elevation in arterial pressure. PMID- 11404277 TI - A construct of interactive feedback control of the GH axis in the male. AB - Growth hormone (GH) secretion is controlled by GH-releasing hormone (GHRH), the GH release-inhibiting hormone somatostatin (SRIF), and autofeedback connections. The ensemble network produces sexually dimorphic patterns of GH secretion. In an effort to formalize this system, we implemented a deterministically based autonomous feedback-driven construct of five principal dose-responsive regulatory interactions: GHRH drive of GH pituitary release, competitive inhibition of GH release by SRIF, GH autofeedback via SRIF with a time delay, delayed GH autonegative feedback on GHRH, and SRIF inhibition of GHRH secretion. This formulation engenders a malelike pattern of successive GH volleys due jointly to positive time-delayed feedback of GH on SRIF and negative feedback of SRIF on GH and GHRH. The multipeak volley is explicated as arising from a reciprocal interaction between GH and GHRH during periods of low SRIF secretion. The applicability of this formalism to neuroendocrine control is explored by initial parameter sensitivity analysis and is illustrated for selected feedback-dependent experimental paradigms. The present construct is not overparameterized and does not require an ad hoc pulse generator to achieve pulsatile GH output. Further evolution of interactive constructs could aid in exploring more complex feedback postulates that confer the vivid sexual dimorphism of female GH profiles. PMID- 11404278 TI - The effect of alcohol consumption on the circadian control of human core body temperature is time dependent. AB - The few controlled studies dealing with the action of alcohol on core body temperature in humans have focused on the effect of a single dose of ethanol and reported that it has a hypothermic effect. No studies report the effects of repeated ethanol intake over a 24-h period, a pattern of consumption much closer to the clinical condition of chronic alcoholism. We therefore designed a trial in which alcohol was repeatedly and regularly administered, with a total dose of 256 g. Nine healthy male volunteers (mean age 23.3 +/- 2.9 yr; range 21-30) each served as his own control. The circadian temperature rhythm was studied by a single-blind, randomized, crossover study that compared a 26-h alcohol session to a 26-h placebo session. The trial controlled for so-called masking effects known to affect temperature. The volunteers were in bed; the ambient temperature was maintained between 20 and 22 degrees C. Meals were standardized. And light was controlled during the night. All sessions took place between November and April. The two sessions were separated by 2 to 5 wk. Rectal temperature was monitored every 20 min throughout the trial. We found the standard hypothermic effect of alcohol in the early hours of the trial, during the daytime, but our principal result is that alcohol consumption induced a very significant hyperthermic effect (+0.36 degrees C) during the night and thereby reduced the circadian amplitude of core body temperature by 43%. The dramatic decrease of the amplitude of circadian temperature rhythm that we observed may explain, at least in part, some clinical signs observed in alcoholic patients, including sleep and mood disorders. We suggest that jet lag, shift work, and aging, which are known to alter body temperature, are aggravated by alcohol consumption. PMID- 11404279 TI - Denervation enhances the physiological effects of the K(ATP) channel during fatigue in EDL and soleus muscle. AB - The objective was to determine whether denervation reduces or enhances the physiological effects of the K(ATP) channel during fatigue in mouse extensor digitorum longus (EDL) and soleus muscle. For this, we measured the effects of 100 microM of pinacidil, a channel opener, and of 10 microM of glibenclamide, a channel blocker, in denervated muscles and compared the data to those observed in innervated muscles from the study of Matar et al. (Matar W, Nosek TM, Wong D, and Renaud JM. Pinacidil suppresses contractility and preserves energy but glibenclamide has no effect during fatigue in skeletal muscle. Am J Physiol Cell Physiol 278: C404-C416, 2000). Pinacidil increased the (86)Rb(+) fractional loss during fatigue, and this effect was 2.6- to 3.4-fold greater in denervated than innervated muscle. Pinacidil also increased the rate of fatigue; for EDL the effect was 2.5-fold greater in denervated than innervated muscle, whereas for soleus the difference was 8.6-fold. A major effect of glibenclamide was an increase in resting tension during fatigue, which was for the EDL and soleus muscle 2.7- and 1.9-fold greater, respectively, in denervated than innervated muscle. A second major effect of glibenclamide was a reduced capacity to recover force after fatigue, an effect observed only in denervated muscle. We therefore suggest that the physiological effects of the K(ATP) channel are enhanced after denervation. PMID- 11404280 TI - Intracellular monocyte and serum cytokine expression is modulated by exhausting exercise and cold exposure. AB - This study tested the hypothesis that exercise elicits monocytic cytokine expression and that prolonged cold exposure modulates such responses. Nine men (age, 24.6 +/- 3.8 y; VO(2 peak), 56.8 +/- 5.6 ml. kg(-1). min(-1)) completed 7 days of exhausting exercise (aerobic, anaerobic, resistive) and underwent three cold, wet exposures (CW). CW trials comprised 200%), and levels of IGF-1 decreased (<55%). The HPD lesion effectively destroyed the entire median eminence [no nerve terminals immunostained for tyrosine hydroxylase (TH) and gonadotropin releasing hormone] and the adjacent arcuate nucleus (no perikarya immunostained for proopiomelanocortin or TH, and no cells expressed neuropeptide Y mRNA). The results support the conclusion that arcuate hypothalamic systems generate long term rhythms in VFI, BW, and energy balance. PMID- 11404282 TI - The influence of nitric oxide synthase 1 on blood flow and interstitial nitric oxide in the kidney. AB - The role of nitric oxide (NO) produced by NO synthase 1 (NOS1) in the renal vasculature remains undetermined. In the present study, we investigated the influence of systemic inhibition of NOS1 by intravenous administration of N(omega)-propyl-L-arginine (L-NPA; 1 mg. kg(-1). h(-1)) and N(5)-(1-imino-3 butenyl)-L-ornithine (v-NIO; 1 mg. kg(-1). h(-1)), highly selective NOS1 inhibitors, on renal cortical and medullary blood flow and interstitial NO concentration in Sprague-Dawley rats. Arterial blood pressure was significantly decreased by administration of both NOS1-selective inhibitors (-11 +/- 1 mmHg with L-NPA and -7 +/- 1 mmHg with v-NIO; n = 9/group). Laser-Doppler flowmetry experiments demonstrated that blood flow in the renal cortex and medulla was not significantly altered following administration of either NOS1-selective inhibitor. In contrast, the renal interstitial level of NO assessed by an in vivo microdialysis oxyhemoglobin-trapping technique was significantly decreased in both the renal cortex (by 36-42%) and medulla (by 32-40%) following administration of L-NPA (n = 8) or v-NIO (n = 8). Subsequent infusion of the nonspecific NOS inhibitor N(omega)-nitro-L-arginine methyl ester (L-NAME; 50 mg. kg(-1). h(-1)) to rats pretreated with either of the NOS1-selective inhibitors significantly increased mean arterial pressure by 38-45 mmHg and significantly decreased cortical (25-29%) and medullary (37-43%) blood flow. In addition, L NAME further decreased NO in the renal cortex (73-77%) and medulla (62-71%). To determine if a 40% decrease in NO could alter renal blood flow, a lower dose of L NAME (5 mg. kg(-1). h(-1); n = 8) was administered to a separate group of rats. The low dose of L-NAME reduced interstitial NO (cortex 39%, medulla 38%) and significantly decreased blood flow (cortex 23-24%, medulla 31-33%). These results suggest that NOS1 does not regulate basal blood flow in the renal cortex or medulla, despite the observation that a considerable portion of NO in the renal interstitial space appears to be produced by NOS1. PMID- 11404283 TI - Activation of alpha(2)-receptors in the rostral ventrolateral medulla evokes natriuresis by a renal nerve mechanism. AB - The contribution of alpha(2)-receptor mechanisms in the rostral ventrolateral medulla (RVLM) in mediating the enhanced renal excretory responses evoked by the intravenous infusion of the alpha(2)-receptor agonist xylazine was examined in ketamine-anesthetized rats. In ketamine-anesthetized rats, the bilateral microinjection of the alpha(2)-receptor antagonist yohimbine into the RVLM significantly reduced the enhanced levels of urine flow rate (V) and urinary sodium excretion (UNaV) produced by xylazine. In contrast, microinjection of yohimbine into the RVLM of chronically bilaterally renal-denervated rats significantly reduced the xylazine-evoked diuretic, but not natriuretic, response. In separate ketamine-anesthetized rats, intravenous xylazine infusion produced a near complete inhibition of renal sympathetic nerve activity (RSNA). The subsequent microinjection of yohimbine into the RVLM reversed this neural response and concurrently decreased V and UNaV. Together, these results indicate that during intravenous infusion, xylazine activates alpha(2)-receptor mechanisms in the RVLM to selectively promote urinary sodium excretion by a renal nerve dependent pathway. In contrast, activation of alpha(2)-receptor in the RVLM affects the renal handling of water by a pathway independent of the renal nerves. This latter pathway may involve an interaction with other brain regions involved in antidiuretic hormone release (e.g., paraventricular nucleus of the hypothalamus). PMID- 11404284 TI - Rectal temperature measurement results in artifactual evidence of selective brain cooling. AB - Selective brain cooling (SBC) is defined as a brain temperature cooler than the temperature of arterial blood from the trunk. Surrogate measures of arterial blood temperature have been used in many published studies on SBC. The use of a surrogate for arterial blood temperature has the potential to confound proper identification of SBC. We have measured brain, carotid blood, and rectal temperatures in conscious sheep exposed to 40, 22, and 5 degrees C. Rectal temperature was consistently higher than arterial blood temperature. Brain temperature was consistently cooler than rectal temperature during all exposures. Brain temperature only fell below carotid blood temperature during the final few hours of 40 degrees C exposure and not at all during the 5 degrees C exposure. Consequently, using rectal temperature as a surrogate for arterial blood temperature does not provide a reliable indication of the status of the SBC effector. We also show that rapid suppression of SBC can result if the animals are disturbed. PMID- 11404285 TI - Metabolic responses to leptin in obese db/db mice are strain dependent. AB - Obese, diabetic C57BL/Ks db/db mice that lack the long-form leptin receptor exhibit no decrease in body weight or food intake when treated with leptin. Here we compared responses to leptin in two strains of db/db mice: C57BL/6J mice that are hyperglycemic and hyperinsulinemic and C57BL/Ks that are hyperglycemic and normo- or hypoinsulinemic. Chronic intraperitoneal infusion of 10 microgram leptin/day partially reversed hyperglycemia in C57BL/6J male mice but exaggerated the diabetic state of female mice. Bolus intraperitoneal injections of 40 microgram leptin/day did not effect glucose in either strain of male db/db mice, whereas chronic intraperitoneal infusion of 20 microgram leptin/day significantly reduced fasting blood glucose in male mice from both strains, especially C57BL/6J mice. Food intake, body weight, rectal temperature, and body fat did not change. Chronic intraperitoneal infusion of 10 microgram leptin/day significantly reduced body fat in lean db/+ C57BL/6J but not in C57BL/Ks mice. Thus peripherally administered leptin is active in mice that have only short-form leptin receptors, and the response is dependent on the method of leptin administration and the background strain. PMID- 11404286 TI - Higher skeletal muscle protein synthesis and lower breakdown after chemotherapy in cachectic mice. AB - The influence of cancer cachexia and chemotherapy and subsequent recovery of skeletal muscle protein mass and turnover was investigated in mice. Cancer cachexia was induced using colon 26 adenocarcinoma, which is characteristic of the human condition, and can be cured with 100% efficacy using an experimental nitrosourea, cystemustine (C(6)H(12)CIN(3)O(4)S). Reduced food intake was not a factor in these studies. Three days after cachexia began, healthy and tumor bearing mice were given a single intraperitoneal injection of cystemustine (20 mg/kg). Skeletal muscle mass in tumor-bearing mice was 41% lower (P < 0.05) than in healthy mice 2 wk after cachexia began. Skeletal muscle wasting was mediated initially by decreased protein synthesis (-38%; P < 0.05) and increased degradation (+131%; P < 0.05); later wasting resulted solely from decreased synthesis (~-54 to -69%; P < 0.05). Acute cytotoxicity of chemotherapy did not appear to have an important effect on skeletal muscle protein metabolism in either healthy or tumor-bearing mice. Recovery began 2 days after treatment; skeletal muscle mass was only 11% lower than in healthy mice 11 days after chemotherapy. Recovery of skeletal muscle mass was affected initially by decreased protein degradation (-80%; P < 0.05) and later by increased protein synthesis (+46 to +73%; P < 0.05) in cured compared with healthy mice. This study showed that skeletal muscle wasted from cancer cachexia and after chemotherapeutic treatment is able to generate a strong anabolic response by making powerful changes to protein synthesis and degradation. PMID- 11404287 TI - Vaginal physiological changes in a model of sexual arousal in anesthetized rats. AB - The understanding of the pathophysiology of female sexual dysfunction suffers from the lack of a convenient model for the study of female genital sexual response. In this study, systemic arterial blood pressure (BP) as well as partial oxygen tension, temperature, and blood engorgement of the vagina [using laser Doppler flowmetry in arbitrary units (AU)] were measured in anesthetized, ovariectomized (1 wk before the start of the experiment) female rats. Vaginal sexual arousal was replicated by electrical stimulation of the pelvic nerve (PNS). PNS induced reproducible increases in the different vaginal parameters (from baseline value, respectively: 16 +/- 10 to 30 +/- 12 mmHg; 34.9 +/- 0.6 to 36 +/- 0.6 degrees C; 450 +/- 196 to 1,500 +/- 360 AU; P < 0.05, paired t-test) and BP (90 +/- 7 to 123 +/- 13 mmHg, P < 0.05, paired t-test). Vaginal vascular resistance was significantly decreased during PNS (from 0.23 +/- 0.15 to 0.08 +/- 0.02 mmHg/AU). Vaginal wall tension was also measured with a force transducer. PNS induced an increase in vaginal wall tension (1.0 +/- 0.2 g), followed by a decrease under the prestimulation value. Intravenous atropine sulfate (1 mg/kg) injection abolished the increase in vaginal wall tension without significantly affecting vaginal vascular resistance. Intravenous vercuronium bromide (2 mg/kg) injection abolished the decrease in vaginal wall tension. Concomitant electrical stimulation of the paravertebral sympathetic chain inhibited vaginal response induced by PNS. Electrical stimulation of the medial preoptic area of the hypothalamus induced a response qualitatively equivalent to PNS with a significant decrease of vaginal vascular resistance. These data support that vaginal contractions involve both smooth and striated muscles and indicate that neural control of vaginal sexual arousal have great similarities in male and female rats. PMID- 11404288 TI - Role of ET(A) receptors in experimental ANG II-induced hypertension in rats. AB - The objectives were to determine if ANG II-induced hypertension is maintained by activation of endothelin type A (ET(A)) receptors by endogenous ET-1 and if this effect is influenced by salt intake. Male rats were maintained on high sodium intake (HS; 6 meq/day) or on normal sodium intake (NS; 2 meq/day). Hypertension was produced by intravenous infusion of ANG II (5 ng/min) for 15 days. Five-day oral dosing with the selective ET(A)-receptor antagonist ABT-627 (~2 mg. kg(-1). day(-1)) reduced mean arterial pressure (MAP) to baseline levels in rats on HS receiving ANG II infusion, but it did not affect MAP in normotensive HS controls. In rats on NS, ABT-627 only transiently decreased MAP in rats receiving ANG II and slightly reduced MAP in normotensive controls. ABT-627 produced mild retention of sodium and water in NS rats receiving ANG II, but not in any other group. These results indicate that ET-1 plays a role in ANG II-induced hypertension via activation of ET(A) receptors and that this role is more prominent in rats on HS. PMID- 11404289 TI - Lengthening contractions are not required to induce protection from contraction induced muscle injury. AB - We tested the hypothesis that lengthening contractions and subsequent muscle fiber degeneration and/or regeneration are required to induce exercise-associated protection from lengthening contraction-induced muscle injury. Extensor digitorum longus muscles in anesthetized mice were exposed in situ to repeated lengthening contractions, isometric contractions, or passive stretches. Three days after lengthening contractions, maximum isometric force production was decreased by 55%, and muscle cross sections contained a significant percentage (18%) of injured fibers. Neither isometric contractions nor passive stretches induced a deficit in maximum isometric force or a significant number of injured fibers at 3 days. Two weeks after an initial bout of lengthening contractions, a second identical bout produced a force deficit (19%) and a percentage of injured fibers (5%) that was smaller than those for the initial bout. Isometric contractions and passive stretches also provided protection from lengthening contraction-induced injury 2 wk later (force deficits = 35 and 36%, percentage of injured fibers = 12 and 10%, respectively), although the protection was less than that provided by lengthening contractions. These data indicate that lengthening contractions and fiber degeneration and/or regeneration are not required to induce protection from lengthening contraction-induced injury. PMID- 11404290 TI - Role of the hepatic function in the development of the pyrogenic tolerance to muramyl dipeptide. AB - We have demonstrated that the hepatic function may have an important role in the development of tolerance to the pyrogenic effect induced by endotoxin. To further investigate if the role of the hepatic function in the development of tolerance also extends to that induced by other pyrogenic stimuli, we investigated the effect of galactosamine, a specific inhibitor of the hepatic protein synthesis, on the development of tolerance to the pyrogenic effect induced by muramyl dipeptide (MDP) in rats. Pyrogenic tolerance was observed after the second intravenous or intraperitoneal injection of MDP (500 microgram/kg), 24 h after the first injection, similar to what was observed with endotoxin. Pyrogenic tolerance was abolished when galactosamine (300 mg/kg ip) was injected simultaneously with MDP (500 microgram/kg iv) on the first day. When uridine (600 mg/kg ip) was administered simultaneously with galactosamine (300 mg/kg ip) and the first injection of MDP (500 microgram/kg ip), pyrogenic tolerance was again observed after the second injection of the peptidoglycan. In conclusion, the hepatic function may not be important only for the development of tolerance to endotoxin, but also to a totally different pyrogenic stimulus such as MDP. PMID- 11404291 TI - Sleep is differently modulated by basal forebrain GABA(A) and GABA(B) receptors. AB - There is evidence that GABA plays a major role in sleep regulation. GABA(A) receptor agonists and different compounds interacting with the GABA(A) receptor complex, such as barbiturates and benzodiazepines, can interfere with the sleep/wake cycle. On the other hand, there is very little information about the possible role of GABA(B) receptors in sleep modulation. The nucleus basalis of Meynert (NBM), a cholinergic area in the basal forebrain, plays a pivotal role in the modulation of sleep and wakefulness, and both GABA(A) and GABA(B) receptors have been described within the NBM. This study used unilateral infusions in the NBM to determine the effects of 3-hydroxy-5-aminomethylisoxazole hydrobromide (muscimol hydrobromide, a GABA(A) receptor subtype agonist) and beta (aminomethyl)-4-chlorobenzenepropanoic acid (baclofen, a GABA(B) receptor subtype agonist) on sleep parameters in freely moving rats by means of polygraphic recordings. Muscimol (0.5 nmol) and baclofen (0.7 nmol) induced an increase in slow-wave sleep and an inhibition of wakefulness. Muscimol, but not baclofen, also caused a decrease in desynchronized sleep parameters. The results reported here indicate that 1) the NBM activation of both GABA(A) and GABA(B) receptors influences the sleep/wake cycle, and 2) GABA(A) but not GABA(B) receptors are important for desynchronized sleep modulation, suggesting that the two GABAergic receptors play different roles in sleep modulation. PMID- 11404292 TI - Effect of a myocardial volume overload on lactate transport in skeletal muscle sarcolemmal vesicles. AB - This study sought to determine the effect of a myocardial volume overload (MVO) on sarcolemmal (SL) lactate (La(-)) transport and the aerobic profile of skeletal muscle. SL vesicles were obtained from female rats 10 wk after either a MVO was induced by creation of an infrarenal fistula (n = 10), or sham surgeries were performed (n = 11). Influx of (14)C-labeled L(+)-La(-) was measured at various unlabeled La(-) concentrations under zero-trans conditions. La(-) transport kinetics were determined using a Michaelis-Menten equation with an added linear component to discriminate between carrier-mediated and diffusional transport. Although heart and lung weights were significantly increased (P < 0.0001) in the MVO group, left ventricular function was only modestly altered (P < 0.05). A significant reduction in type I myosin heavy chain (MHC) in the soleus and a strong trend (P = 0.06) for a reduced type IIx MHC in the plantaris were observed in MVO rats, but no differences in citrate synthase activity or monocarboxylate transporter proteins (MCT)-1 expression were noted in any muscle. Carrier mediated La(-) influx into SL vesicles was similar between sham and MVO (K(m) = 12 +/- 1 and 18 +/- 3 mM; apparent V(max) = 772 +/- 99 and 827 +/- 80 nmol. mg( 1). min(-1), respectively). Total influx at 100 mM was lower in MVO, and this was due to a 30% reduction in membrane diffusion. In conclusion, a 10-wk MVO did not alter MCT-mediated La(-) transport or protein expression but was associated with modest changes in myofibrillar proteins and impaired SL diffusive properties. PMID- 11404293 TI - Reduced neuromuscular activity and force generation during prolonged cycling. AB - We examined neuromuscular activity during stochastic (variable intensity) 100-km cycling time trials (TT) and the effect of dietary carbohydrate manipulation. Seven endurance-trained cyclists performed two 100-km TT that included five 1-km and four 4-km high-intensity epochs (HIE) during which power output, electromyogram (EMG), and muscle glycogen data were analyzed. The mean power output of the 4-km HIE decreased significantly throughout the trial from 319 +/- 48 W for the first 4-km HIE to 278 +/- 39 W for the last 4-km HIE (P < 0.01). The mean integrated EMG (IEMG) activity during the first 4-km HIE was 16.4 +/- 9.8% of the value attained during the pretrial maximal voluntary contraction (MVC). IEMG decreased significantly throughout the trial, reaching 11.1 +/- 5.6% during the last 4-km HIE (P < 0.01). The study establishes that neuromuscular activity in peripheral skeletal muscle falls parallel with reduction in power output during bouts of high-intensity exercise. These changes occurred when <20% of available muscle was recruited and suggest the presence of a central neural governor that reduces the active muscle recruited during prolonged exercise. PMID- 11404294 TI - Phase-advance shifts of human circadian pacemaker are accelerated by daytime physical exercise. AB - Effects of forced sleep-wake schedules with and without physical exercise were examined on the human circadian pacemaker under dim light conditions. Subjects spent 15 days in an isolation facility separately without knowing the time of day and followed a forced sleep-wake schedule of a 23 h 40-min period for 12 cycles, and physical exercise was imposed twice per waking period for 2 h each with bicycle- or rowing-type ergometers. As a result, plasma melatonin rhythm was significantly phase advanced with physical exercise, whereas it was not changed without exercise. The difference in phase was already significant 6 days after the start of exercise. The amplitude of melatonin rhythm was not affected. A single pulse of physical exercise in the afternoon or at midnight significantly phase delayed the melatonin rhythms when compared with the prepulse phase, but the amount of phase shift was not different from that observed in the sedentary controls. These findings indicate that physical exercise accelerates phase advance shifts of the human circadian pacemaker associated with the forced sleep wake schedule. PMID- 11404295 TI - Dynamic relationship between sympathetic nerve activity and renal blood flow: a frequency domain approach. AB - Blood pressure displays an oscillation at 0.1 Hz in humans that is well established to be due to oscillations in sympathetic nerve activity (SNA). However, the mechanisms that control the strength or frequency of this oscillation are poorly understood. The aim of the present study was to define the dynamic relationship between SNA and the vasculature. The sympathetic nerves to the kidney were electrically stimulated in six pentobarbital-sodium anesthetized rabbits, and the renal blood flow response was recorded. A pseudo-random binary sequence (PRBS) was applied to the renal nerves, which contains equal spectral power at frequencies in the range of interest (<1 Hz). Transfer function analysis revealed a complex system composed of low-pass filter characteristics but also with regions of constant gain. A model was developed that accounted for this relationship composed of a 2 zero/4 pole transfer function. Although the position of the poles and zeros varied among animals, the model structure was consistent. We also found the time delay between the stimulus and the RBF responses to be consistent among animals (mean 672 +/- 22 ms). We propose that the identification of the precise relationship between SNA and renal blood flow (RBF) is a fundamental and necessary step toward understanding the interaction between SNA and other physiological mediators of RBF. PMID- 11404296 TI - Beat-to-beat cardiovascular responses to rapid, low-level LBNP in humans. AB - The hypothesis tested was that there are significant transient changes in the cardiovascular variables after rapid onset and release of mild lower body negative pressure (LBNP, -20 mmHg), even in experimental situations where there is no detectable change in steady-state values. Twelve subjects participated in the study. Heart rate, stroke volume (SV), cardiac output, mean arterial pressure (MAP), total peripheral resistance (TPR), acral and nonacral skin blood flow, and blood flow velocity in the brachial artery were continuously recorded during the pre-LBNP period (0-120 s), during LBNP (120-420 s), and during the post-LBNP period (420-600 s). The main finding was that MAP is transiently but strongly affected by rapid changes in LBNP as small as -20 mmHg. There was also a characteristic asymmetry in cardiovascular responses to the onset and release of LBNP, particularly in the responses in SV. The transient changes in MAP indicate that the neural responses that affect TPR are not fast enough to compensate for the rapid changes in LBNP. In this case, the arterial baroreceptors will be activated as well as the low-pressure baroreceptors that sense central venous pressure. This must be taken into consideration in future discussions of the results of LBNP protocols. PMID- 11404297 TI - Postprandial neuronal activation in the nucleus of the solitary tract is partly mediated by CCK-A receptors. AB - CCK-A receptors and neurons of the nucleus of the solitary tract (NTS) are involved in the regulation of food intake, and in rats, there is evidence for involvement of an intestinal vagal afferent pathway. Studies investigating the role of CCK-A receptors in activation of NTS neurons using highly selective CCK-A receptor agonists and antagonists have yielded conflicting data. In the present study, we investigated CCK-induced and postprandial activation of NTS neurons, together with food intake studies, in CCK-A receptor-deficient Otsuka Long-Evans Tokushima fatty (OLETF) rats. Activated NTS neurons were detected using immunohistological staining for c-Fos protein. Exogenous CCK increased the number of c-Fos protein-positive cells in the NTS of Sprague-Dawley and CCK-A receptor intact Long-Evans Tokushima Otsuka (LETO) rats but had no effect in CCK-A receptor-deficient OLETF rats. Food intake-induced c-Fos protein expression in NTS neurons was significantly reduced in CCK-A receptor-deficient OLETF rats compared with Sprague-Dawley or LETO rats. Postprandial c-Fos protein expression in the NTS was also significantly decreased after pretreatment with the CCK-A receptor antagonist MK329 after both short- and long-term fasting periods. Exogenous CCK decreased cumulative food intake in Sprague-Dawley and LETO rats but not in OLETF rats. These data demonstrate that CCK-A receptors are involved in the CCK- and food-induced c-Fos protein expression in the NTS. Taken together with the results of the food intake studies, this suggests that activation of CCK A receptors is involved in the postprandial activation of NTS neurons and in the regulation of food intake. PMID- 11404298 TI - Small bowel motility and colonic transit are altered in dogs with moderate renal failure. AB - Although gastrointestinal complications are common in patients with renal disease, the effects of renal dysfunction on bowel motility and gut transit times are not well known. We assessed gastrointestinal electromyographic activity, gastric emptying rate, orocolonic transit time, oroanal transit time, and xylose absorption before and after surgically inducing a 66% decrease in glomerular filtration rate in dogs. Moderate renal failure induced no gross or microscopic gastrointestinal lesions but caused a 16-42% increase in gastrointestinal motility indexes. We found a 24% decrease in the propagation velocity of the myoelectrical migrating complex in the duodenojejunal segment, a 30% decrease in phase I duration in duodenal and jejunal regions, a 20% increase in the total irregular electrical activity of the small intestine, and a 22% increase in duration of the meal response in the duodenum and jejunum. Renal failure did not change xylose absorption, gastric emptying rate, and orocolonic transit time but decreased colonic transit time by 38%. The mean weight of feces was increased. These results indicate that moderate renal failure alters duodenojejunal motility and decreases colonic transit time. PMID- 11404299 TI - Renal interstitial hydrostatic pressure and sodium excretion during acute volume expansion in diabetic rats. AB - Experiments were performed to test the hypothesis that the renal interstitial hydrostatic pressure (RIHP) response to acute volume expansion is suppressed in diabetes mellitus. Sprague-Dawley rats received streptozotocin (STZ rats; 65 mg/kg ip) or vehicle (Sham rats). Two weeks later, RIHP and Na(+) excretion responses to acute graded volume expansion with isotonic saline were quantified under Inactin anesthesia (0.1 mg/kg ip). In Sham rats, acute graded volume expansion to 10% body wt produced increases in RIHP (Delta = 12.2 +/- 2.4 mmHg), urine flow (Delta = 54 +/- 8 microliter. min(-1). g(-1)), and Na(+) excretion (Delta = 11.5 +/- 1.9 mueq. min(-1). g(-1)). In STZ rats, these volume expansion induced responses were significantly blunted (RIHP by 50%, urine flow by 81%, and Na(+) excretion by 76%). Renal decapsulation eliminated the differences between STZ and Sham rats with regard to volume expansion-induced increases in RIHP, urine flow, and Na(+) excretion. Renal denervation normalized the RIHP response to volume expansion and improved the diuretic and natriuretic responses in STZ rats. Moreover, diuretic and natriuretic responses to direct changes in RIHP (induced by renal interstitial volume expansion) were blunted in STZ rats. We conclude that diminished alterations in RIHP, as well as a reduced impact of RIHP on Na(+) excretion, contribute to the impaired diuretic and natriuretic responses to acute volume expansion during the early stage of diabetes. PMID- 11404300 TI - Arachidonate dilates basilar artery by lipoxygenase-dependent mechanism and activation of K(+) channels. AB - Dilatation of cerebral arterioles in response to arachidonic acid is dependent on activity of cyclooxygenase. In this study, we examined mechanisms that mediate dilatation of the basilar artery in response to arachidonate. Diameter of the basilar artery (baseline diameter = 216 +/- 7 micrometer) (means +/- SE) was measured using a cranial window in anesthetized rats. Arachidonic acid (10 and 100 microM) produced concentration-dependent vasodilatation that was not inhibited by indomethacin (10 mg/kg iv) or N(G)-nitro-L-arginine (100 microM) but was inhibited markedly by baicalein (10 micrometerM) or nordihydroguaiaretic acid (NDGA; 10 microM), inhibitors of the lipoxygenase pathway. Dilatation of the basilar artery was also inhibited markedly by tetraethylammonium ion (TEA; 1 mM) or iberiotoxin (50 nM), inhibitors of calcium-dependent potassium channels. For example, 10 microM arachidonate dilated the basilar artery by 19 +/- 7 and 1 +/- 1% in the absence and presence of iberiotoxin, respectively. Measurements of membrane potential indicated that arachidonate produced hyperpolarization of the basilar artery that was blocked completely by TEA. Incubation with [(3)H]arachidonic acid followed by reverse-phase and chiral HPLC indicated that the basilar artery produces relatively small quantities of prostanoids but large quantities of 12(S)-hydroxyeicosatetraenoic acid (12-S-HETE), a lipoxygenase product. Moreover, the production of 12-HETE was inhibited by baicalein or NDGA. These findings suggest that dilatation of the basilar artery in response to arachidonate is mediated by a product(s) of the lipoxygenase pathway, with activation of calcium-dependent potassium channels and hyperpolarization of vascular muscle. PMID- 11404301 TI - A role for NPY overexpression in the dorsomedial hypothalamus in hyperphagia and obesity of OLETF rats. AB - Otsuka Long-Evans Tokushima Fatty (OLETF) rats lacking CCK-A receptors are hyperphagic, obese, and diabetic. We have previously demonstrated that these rats have a peripheral satiety deficit resulting in increased meal size. To examine the potential role of hypothalamic pathways in the hyperphagia and obesity of OLETF rats, we compared patterns of hypothalamic neuropeptide Y (NPY), proopiomelanocortin (POMC), and leptin receptor mRNA expression in ad libitum-fed Long-Evans Tokushima (LETO) and OLETF rats and food-restricted OLETF rats that were pair-fed to the intake of LETO controls. Pair feeding OLETF rats prevented their increased body weight and elevated levels of plasma insulin and leptin and normalized their elevated POMC and decreased NPY mRNA expression in the arcuate nucleus. In contrast, NPY expression was upregulated in the dorsomedial hypothalamus (DMH) in pair-fed OLETF rats. A similar DMH NPY overexpression was evident in 5-wk-old preobese OLETF rats. These findings suggest a role for DMH NPY upregulation in the etiology of OLETF hyperphagia and obesity. PMID- 11404302 TI - Effect of in vivo fetal infusion of dexamethasone at 0.75 GA on fetal ovine resistance artery responses to ET-1. AB - At 110-111 days gestation, instrumented fetal sheep were administered saline or dexamethasone (2.2 microgram. kg(-1). h(-1) iv) for 48 h. Measurement of fetal blood pressure showed a greater increase in dexamethasone-treated (n = 6) compared with control (n = 5) fetuses (7.3 +/- 2.3 vs. 0.6 +/- 2.3 mmHg, P < 0.05). Fetuses were delivered by cesarean section, and the femoral muscle and brain were obtained under halothane anesthesia. Femoral and middle cerebral arteries (approximately 320-micrometer internal diameter) were evaluated using wire myography. Sensitivity to KCl (2.5-125 mM) and the magnitude of the maximal vasoconstriction to 125 mM K(+) were similar in femoral and middle cerebral arteries from dexamethasone-treated vs. control fetuses. Acetylcholine-induced vasorelaxation was similar in femoral arteries from control and dexamethasone treated fetuses. Middle cerebral arteries did not relax to acetylcholine. Sensitivity to endothelin-1 (ET-1; 0.1 pM-0.1 microM) and magnitude of the ET-1 induced vasoconstriction were greater in femoral arteries from dexamethasone treated vs. control fetuses (P < 0.05). Autoradiographical studies with receptor specific ligands demonstrated increased ET(A)-receptor binding, the principal receptor subtype, in femoral muscle vessels (P < 0.001) but decreased ET(A) receptor binding in middle cerebral arteries (P < 0.01) from dexamethasone treated compared with control fetuses. Relatively little ET(B)-receptor binding was evident in all tissues examined. We conclude that hyperreactivity to ET-1, due to increased ET(A)-receptor binding, may be involved in the dexamethasone induced increase in peripheral vascular resistance in fetal sheep in vivo. PMID- 11404303 TI - Central modulation of the NO/cGMP pathway affects the MPOA-induced intracavernous pressure response. AB - Alterations in the nitric oxide (NO)/cGMP levels in hypothalamic nuclei, including the medial preoptic area (MPOA), regulate critical aspects of sexual behavior and penile reflexes. However, the effects of altered central nervous system (CNS) NO/cGMP levels at the end organ level, that is, on the magnitude/quality of the erection so achieved [intracavernous pressure (ICP) response], has yet to be evaluated. The goal of this report was to evaluate the effects of intrathecal administration of modulators of NO and cGMP levels on ICP responses to stimulation of the MPOA and cavernous nerve in rats in vivo. In all cases, intrathecal administration of compounds that increase and decrease cGMP and NO levels, respectively, was associated with corresponding increases and decreases in the MPOA-stimulated ICP response. Specifically, sodium nitroprusside (SNP), 8-bromo-cGMP, and sildenafil increased the MPOA-stimulated ICP response, whereas N(omega)-nitro-L-arginine methyl ester reduced it. None of the intrathecal treatments had detectable effects on blood pressure or the cavernous nerve-stimulated ICP response, although intravenous sildenafil increased the latter. These data clearly indicate that intrathecal drug administration affects central and not peripheral neural mechanisms and, moreover, documents that CNS NO/cGMP levels can affect erectile capacity per se (i.e., ICP) in the rat model. PMID- 11404304 TI - Nitric oxide modulates renal sensory nerve fibers by mechanisms related to substance P receptor activation. AB - Nerve terminals containing neuronal nitric oxide synthase (nNOS) are localized in the renal pelvic wall where the sensory nerves containing substance P and calcitonin gene-related peptide (CGRP) are found. We examined whether nNOS is colocalized with substance P and CGRP. All renal pelvic nerve fibers that contained nNOS-like immunoreactivity (-LI) also contained substance P-LI and CGRP LI. In anesthetized rats, renal pelvic perfusion with the nNOS inhibitor S-methyl L-thiocitrulline (L-SMTC, 20 microM) prolonged the afferent renal nerve activity (ARNA) response to a 3-min period of increased renal pelvic pressure from 5 +/- 0.4 to 21 +/- 2 min (P < 0.01, n = 14). The magnitude of the ARNA response was unaffected by L-SMTC. Similar effects were produced by N(omega)-nitro-L-arginine methyl ester (L-NAME) but not D-NAME. Increasing renal pelvic pressure produced similar increases in renal pelvic release of substance P before and during L SMTC, from 5.9 +/- 1.4 to 13.6 +/- 4.2 pg/min before and from 4.9 +/- to 12.6 +/- 2.7 pg/min during L-SMTC. L-SMTC also prolonged the ARNA response to renal pelvic perfusion with substance P (3 microM) from 1.2 +/- 0.2 to 5.6 +/- 1.1 min (P < 0.01, n = 9) without affecting the magnitude of the ARNA response. IN CONCLUSION: activation of NO may function as an inhibitory neurotransmitter regulating the activation of renal mechanosensory nerve fibers by mechanisms related to activation of substance P receptors. PMID- 11404305 TI - Factors that increase the contractile tone of the ductus arteriosus also regulate its anatomic remodeling. AB - Permanent closure of the full-term newborn ductus arteriosus (DA) occurs only if profound hypoxia develops within the vessel wall during luminal obliteration. We used fetal and newborn baboons and lambs to determine why the immature DA fails to remodel after birth. When preterm newborns were kept in a normoxic range (Pa(O(2)): 50-90 mmHg), 86% still had a small patent DA on the sixth day after birth; in addition, the preterm DA wall was only mildly hypoxic and had only minimal remodeling. The postnatal increase in Pa(O(2)) normally induces isometric contractile responses in rings of DA; however, the excessive inhibitory effects of endogenous prostaglandins and nitric oxide, coupled with a weaker intrinsic DA tone, make the preterm DA appear to have a smaller increment in tension in response to oxygen than the DA near term. We found that oxygen concentrations, beyond the normoxic range, produce an additional increase in tension in the preterm DA that is similar to the contractile response normally seen at term. We predicted that preterm newborns, kept at a higher Pa(O(2)), would have increased DA tone and would be more likely to obliterate their lumen. We found that preterm newborns, maintained at a Pa(O(2)) >200 mmHg, had only a 14% incidence of patent DA. Even though DA constriction was due to elevated Pa(O(2)), obliteration of the lumen produced profound hypoxia of the DA wall and the same features of remodeling that were observed at term. DA wall hypoxia appears to be both necessary and sufficient to produce anatomic remodeling in preterm newborns. PMID- 11404306 TI - Early-age heat exposure affects skeletal muscle satellite cell proliferation and differentiation in chicks. AB - Exposure of young chicks to thermal conditioning (TC; i.e., 37 degrees C for 24 h) resulted in significantly improved body and muscle growth at a later age. We hypothesized that TC causes an increase in satellite cell proliferation, necessary for further muscle hypertrophy. An immediate increase was observed in satellite cell DNA synthesis in culture and in vivo in response to TC of 3-day old chicks to levels that were significantly higher than those of control chicks. This was accompanied by a marked induction of insulin-like growth factor-I (IFG I), but not hepatocyte growth factor in the breast muscle. No significant difference between treatments in plasma IGF-I levels was observed. A marked elevation in muscle regulatory factors on day 5, followed by a decline in cell proliferation on day 6 together with continuous high levels of IGF-I in the TC chick muscle may indicate accelerated cell differentiation. These data suggest a central role for IGF-I in the immediate stimulation of satellite cell myogenic processes in response to heat exposure. PMID- 11404307 TI - Fluid volume and osmoregulation in humans after a week of head-down bed rest. AB - Body fluid homeostasis was investigated during chronic bed rest (BR) and compared with that of acute supine conditions. The hypothesis was tested that 6 degrees head-down BR leads to hypovolemia, which activates antinatriuretic mechanisms so that the renal responses to standardized saline loading are attenuated. Isotonic (20 ml/kg body wt) and hypertonic (2.5%, 7.2 ml/kg body wt) infusions were performed in eight subjects over 20 min following 7 and 10 days, respectively, of BR during constant sodium intake (200 meq/day). BR decreased body weight (83.0 +/ 4.8 to 81.8 +/- 4.4 kg) and increased plasma osmolality (285.9 +/- 0.6 to 288.5 +/- 0.9 mosmol/kgH(2)O, P < 0.05). Plasma ANG II doubled (4.2 +/- 1.2 to 8.8 +/- 1.8 pg/ml), whereas other endocrine variables decreased: plasma atrial natriuretic peptide (42 +/- 3 to 24 +/- 3 pg/ml), urinary urodilatin excretion rate (4.5 +/- 0.3 to 3.2 +/- 0.1 pg/min), and plasma vasopressin (1.7 +/- 0.3 to 0.8 +/- 0.2 pg/ml, P < 0.05). During BR, the natriuretic response to the isotonic saline infusion was augmented (39 +/- 8 vs. 18 +/- 6 meq sodium/350 min), whereas the response to hypertonic saline was unaltered (32 +/- 8 vs. 29 +/- 5 meq/350 min, P < 0.05). In conclusion, BR elicits antinatriuretic endocrine signals, but it does not attenuate the renal natriuretic response to saline stimuli in men; on the contrary, the response to isotonic saline is augmented. PMID- 11404308 TI - Chronic effect of insulin-like growth factor I on renin synthesis, secretion, and renal function in fetal sheep. AB - In the adult, insulin-like growth factor I (IGF-I) increases glomerular filtration rate (GFR) and renal blood flow (RBF) during both acute and chronic treatment. To study its effects on the developing kidney, chronically catheterized fetal sheep (120 +/- 1 days gestation) were infused intravenously for up to 10 days with 80 microgram/h IGF-I (n = 5) or vehicle (0.1% BSA in saline, n = 6). In contrast to previous acute studies in adult rats and humans, after 4 h of IGF-I fetal GFR and RBF were unchanged. Fractional sodium reabsorption increased (P < 0.05). However, by 4 days, GFR per kilogram had risen by 35 +/- 13% (P < 0.05), whereas RBF remained unchanged. Tubular growth and maturation may have occurred, as proximal tubular sodium reabsorption increased by ~35% (P < 0.005). Therefore, despite a marked increase in filtered sodium (~30%, P < 0.05), fractional sodium reabsorption did not change. Although the effects of IGF-I on renal function were delayed, plasma renin activity and concentration were both elevated after 4 h and remained high at 4 days (P < 0.05). Despite this, arterial pressure and heart rate did not change. Kidneys of IGF-I-infused fetuses weighed ~30% more (P = 0.05) and contained ~75% more renin than control fetuses (P < 0.005). Thus, in the fetus, the renal effects of long term IGF-I infusion are very different from the adult, possibly because IGF-I stimulated kidney growth. PMID- 11404309 TI - Pulmonary surfactant in birds: coping with surface tension in a tubular lung. AB - As birds have tubular lungs that do not contain alveoli, avian surfactant predominantly functions to maintain airflow in tubes rather than to prevent alveolar collapse. Consequently, we have evaluated structural, biochemical, and functional parameters of avian surfactant as a model for airway surfactant in the mammalian lung. Surfactant was isolated from duck, chicken, and pig lung lavage fluid by differential centrifugation. Electron microscopy revealed a uniform surfactant layer within the air capillaries of the bird lungs, and there was no tubular myelin in purified avian surfactants. Phosphatidylcholine molecular species of the various surfactants were measured by HPLC. Compared with pig surfactant, both bird surfactants were enriched in dipalmitoylphosphatidylcholine, the principle surface tension-lowering agent in surfactant, and depleted in palmitoylmyristoylphosphatidylcholine, the other disaturated phosphatidylcholine of mammalian surfactant. Surfactant protein (SP) A was determined by immunoblot analysis, and SP-B and SP-C were determined by gel filtration HPLC. Neither SP-A nor SP-C was detectable in either bird surfactant, but both preparations of surfactant contained SP-B. Surface tension function was determined using both the pulsating bubble surfactometer (PBS) and capillary surfactometer (CS). Under dynamic cycling conditions, where pig surfactant readily reached minimal surface tension values below 5 mN/m, neither avian surfactant reached values below 15 mN/m within 10 pulsations. However, maximal surface tension of avian surfactant was lower than that of porcine surfactant, and all surfactants were equally efficient in the CS. We conclude that a surfactant composed primarily of dipalmitoylphosphatidylcholine and SP-B is adequate to maintain patency of the air capillaries of the bird lung. PMID- 11404310 TI - Effect of nitric oxide in the nucleus isthmi on the hypoxic and hypercarbic drive to breathing of toads. AB - Nucleus isthmi (NI) is a mesencephalic structure of the amphibian brain that has been reported to participate in CO(2) chemoreception and in the ventilatory response to hypoxia. However, no information exists about the modulators and/or mediators involved. In the present study, we assessed the participation of nitric oxide (NO) in the hypoxic and hypercarbic drive to breathing, specifically in the NI. We compared the ventilatory and cardiovascular responses with hypoxia and hypercarbia after microinjecting 100 nmol/0.5 microliter of N(G)-nitro-L-arginine methyl ester (L-NAME; an NO synthase blocker) into the NI of toads (Bufo paracnemis). L-NAME had no effect under resting conditions. Hypoxia elicited an increase in ventilation in control and vehicle toads by elevating tidal volume (V(T)). Hypercarbia caused hyperventilation in all groups due to an increase in both V(T) and frequency. The microinjection of L-NAME into the NI elicited an increase in ventilatory response to hypoxia and hypercarbia due to a higher V(T.) We conclude that NO in the NI has an inhibitory effect when the respiratory drive is high, acting on V(T). PMID- 11404311 TI - Role of plasma ANG II in the excretion of acute sodium load in a bird with salt glands (Anas platyrhynchos). AB - This study was designed to further examine the role of plasma ANG II in the excretion of sodium in the Pekin duck, a bird with salt glands. Renal and extrarenal (salt gland) excretion of an intravenously administered isotonic saline load was monitored over a 4-h period in a group of eight birds under two conditions: the control condition, in which isotonic saline infusion decreased endogenous plasma ANG II from 102.6 to 16.5 pg/ml, and the experimental condition, in which ANG II suppression was prevented by intravenous infusion of a 3.5 ng. kg(-1). min(-1) dose of synthetic ANG II. ANG II infusion significantly decreased the total sodium excretion (by 15%), primarily via an inhibition of salt gland output. The results suggest that ANG II suppression facilitates the excretion of an administered sodium load in birds with salt glands. PMID- 11404312 TI - Interaction of NaCl and behavioral stress on endogenous sodium pump ligands in rats. AB - Our study investigated the hypothesis that the combination of a high NaCl diet and social isolation stress would increase systolic blood pressure (SBP) and endogenous sodium pump ligands (SPL), ouabainlike compound (OLC), and marinobufagenin (MBG). Excretion of MBG and OLC, SBP, and organ weights were studied in four groups (n = 8) of male Fisher 344 x Norwegian brown rats: controls, socially isolated (Iso), 4% NaCl diet (Salt), and the combination of Salt and Iso (Iso+Salt). In Salt, MBG excretion increased by 78% (P < 0.01), whereas SBP and OLC remained unchanged. In Iso, SBP and MBG did not change, but OLC peaked on day 1. In the Iso+Salt, SBP increased by 9 mmHg, MBG excretion increased (42.0 +/- 7.6 vs. 10.0 +/- 1.5 pmol/24 h, P < 0.01), whereas OLC peaked at day 1 (25.0 +/- 2.5 vs. 10.0 +/- 2.0 pmol/24 h, P < 0.01) and remained elevated. Heart and kidney weights were increased in Salt and Iso+Salt. Aortic weights were increased in Iso and Iso+Salt. Thus a high NaCl intake stimulates MBG excretion, whereas isolation stress stimulates OLC. The combination of Salt and Iso is accompanied by marked stimulation of both SPL. PMID- 11404313 TI - Heat shock prevents simulated ischemia-induced apoptosis in renal tubular cells via a PKC-dependent mechanism. AB - Heat shock produces cellular tolerance to a variety of adverse conditions; however, the protective effect of heat shock on renal cell ischemic injury remains unclear. Protein kinase C (PKC) has been implicated in the signaling mechanisms of acute preconditioning, yet it remains unknown whether PKC mediates heat shock-induced delayed preconditioning in renal cells. To study this, renal tubular cells (LLC-PK1) were exposed to thermal stress (43 degrees C) for 1 h and heat shock protein (HSP) 72 induction was confirmed by Western blot analysis. Cells were subjected to simulated ischemia 24 h after thermal stress, and the effect of heat shock (delayed preconditioning) on ischemia-induced apoptosis (terminal deoxynucleotidyl transferase dUTP nick-end labeling) and B cell lymphoma 2 (Bcl(2)) expression (Western) was determined. Subsequently, the effect of PKC inhibition on HSP72 induction and heat stress-induced ischemic tolerance was evaluated. Thermal stress induced HSP72 production, increased Bcl(2) expression, and prevented simulated ischemia-induced renal tubular cell apoptosis. PKC inhibition abolished thermal induction of HSP72 and prevented heat stress-induced ischemic tolerance. These data demonstrate that thermal stress protects renal tubular cells from simulated ischemia-induced apoptosis through a PKC-dependent mechanism. PMID- 11404314 TI - Moderate levels of ethanol induce expression of vascular endothelial growth factor and stimulate angiogenesis. AB - Alcohol abuse has a negative impact on human health; however, epidemiological studies show that moderate consumption of ethanol (EtOH) reduces the risk of coronary heart disease, sudden cardiac death, and ischemic stroke. The mechanisms for these reductions in cardiovascular disease are not well established. Using cultured coronary artery vascular smooth muscle cells, we found that moderate levels of EtOH (10 and 20 mM) caused dose-related increases in both vascular endothelial growth factor (VEGF) mRNA (Northern blot) expression (1.9- and 2.6 fold) and VEGF protein (ELISA) expression (19 and 68%) compared with control (P < 0.05). EtOH at 0.25 g. kg(-1). day(-1) (7 days) increased VEGF mRNA expression by 1.48-fold over control, and increased vessel length density from 3.9 +/- 0.7 (control) to 6.0 +/- 0.3 mm/mm(2) (P < 0.05) in chick chorioallantoic membrane (CAM). We conclude that moderate levels of ethanol can induce VEGF expression and stimulate angiogenesis in chick CAM. Therefore, the results provide a theoretical basis for speculating that the cardiovascular-protective effects of moderate alcohol consumption may be partly mediated through VEGF-induced angiogenesis. PMID- 11404315 TI - George Beadle's other hypothesis: one-gene, one-trait. PMID- 11404316 TI - Defining cosQ, the site required for termination of bacteriophage lambda DNA packaging. AB - Bacteriophage lambda is a double-stranded DNA virus that processes concatemeric DNA into virion chromosomes by cutting at specific recognition sites termed cos. A cos is composed of three subsites: cosN, the nicking site; cosB, required for packaging initiation; and cosQ, required for termination of chromosome packaging. During packaging termination, nicking of the bottom strand of cosN depends on cosQ, suggesting that cosQ is needed to deliver terminase to the bottom strand of cosN to carry out nicking. In the present work, saturation mutagenesis showed that a 7-bp segment comprises cosQ. A proposal that cosQ function requires an optimal sequence match between cosQ and cosNR, the right cosN half-site, was tested by constructing double cosQ mutants; the behavior of the double mutants was inconsistent with the proposal. Substitutions in the 17-bp region between cosQ and cosN resulted in no major defects in chromosome packaging. Insertional mutagenesis indicated that proper spacing between cosQ and cosN is required. The lethality of integral helical insertions eliminated a model in which DNA looping enables cosQ to deliver a gpA protomer for nicking at cosN. The 7 bp of cosQ coincide exactly with the recognition sequence for the Escherichia coli restriction endonuclease, EcoO109I. PMID- 11404317 TI - Identification of important amino acid residues that modulate binding of Escherichia coli GroEL to its various cochaperones. AB - Genetic experiments have shown that the GroEL/GroES chaperone machine of Escherichia coli is absolutely essential, not only for bacterial growth but also for the propagation of many bacteriophages including lambda. The virulent bacteriophages T4 and RB49 are independent of the host GroES function, because they encode their own cochaperone proteins, Gp31 and CocO, respectively. E. coli groEL44 mutant bacteria do not form colonies above 42 degrees nor do they propagate bacteriophages lambda, T4, or RB49. We found that the vast majority (40/46) of spontaneous groEL44 temperature-resistant colonies at 43 degrees were due to the presence of an intragenic suppressor mutation. These suppressors define 21 different amino acid substitutions in GroEL, each affecting one of 13 different amino acid residues. All of these amino acid residues are located at or near the hinge, which regulates the large en bloc movements of the GroEL apical domain. All of these intragenic suppressors support bacteriophages lambda, T4, and RB49 growth to various extents in the presence of the groEL44 allele. Since it is known that the GroEL44 mutant protein does not interact effectively with Gp31, the suppressor mutations should enhance cochaperone binding. Analogous intragenic suppressor studies were conducted with the groEL673 temperature sensitive allele. PMID- 11404318 TI - Evolutionary cheating in Escherichia coli stationary phase cultures. AB - Starved cultures of Escherichia coli are highly dynamic, undergoing frequent population shifts. The shifts result from the spread of mutants able to grow under conditions that impose growth arrest on the ancestral population. To analyze competitive interactions underlying this dynamic we measured the survival of a typical mutant and the wild type during such population shifts. Here we show that the survival advantage of the mutant at any given time during a takeover is inversely dependent on its frequency in the population, its growth adversely affects the survival of the wild type, and its ability to survive in stationary phase at fixation is lower than that of its ancestor. These mutants do not enter, or exit early, the nondividing stationary-phase state, cooperatively maintained by the wild type. Thus they end up overrepresented as compared to their initial frequency at the onset of the stationary phase, and subsequently they increase disproportionately their contribution in terms of progeny to the succeeding generation in the next growth cycle, which is a case of evolutionary cheating. If analyzed through the game theory framework, these results might be explained by the prisoner's dilemma type of conflict, which predicts that selfish defection is favored over cooperation. PMID- 11404319 TI - Evidence for two mechanisms of palindrome-stimulated deletion in Escherichia coli: single-strand annealing and replication slipped mispairing. AB - Spontaneous deletion mutations often occur at short direct repeats that flank inverted repeat sequences. Inverted repeats may initiate genetic rearrangements by formation of hairpin secondary structures that block DNA polymerases or are processed by structure-specific endonucleases. We have investigated the ability of inverted repeat sequences to stimulate deletion of flanking direct repeats in Escherichia coli. Propensity for cruciform extrusion in duplex DNA correlated with stimulation of flanking deletion, which was partially sbcD dependent. We propose two mechanisms for palindrome-stimulated deletion, SbcCD dependent and SbcCD independent. The SbcCD-dependent mechanism is initiated by SbcCD cleavage of cruciforms in duplex DNA followed by RecA-independent single-strand annealing at the flanking direct repeats, generating a deletion. Analysis of deletion endpoints is consistent with this model. We propose that the SbcCD-independent pathway involves replication slipped mispairing, evoked from stalling at hairpin structures formed on the single-stranded lagging-strand template. The skew of SbcCD-independent deletion endpoints with respect to the direction of replication supports this hypothesis. Surprisingly, even in the absence of palindromes, SbcD affected the location of deletion endpoints, suggesting that SbcCD-mediated strand processing may also accompany deletion unassociated with secondary structures. PMID- 11404320 TI - RAG4 gene encodes a glucose sensor in Kluyveromyces lactis. AB - The rag4 mutant of Kluyveromyces lactis was previously isolated as a fermentation deficient mutant, in which transcription of the major glucose transporter gene RAG1 was affected. The wild-type RAG4 was cloned by complementation of the rag4 mutation and found to encode a protein homologous to Snf3 and Rgt2 of Saccharomyces cerevisiae. These two proteins are thought to be sensors of low and high concentrations of glucose, respectively. Rag4, like Snf3 and Rgt2, is predicted to have the transmembrane structure of sugar transporter family proteins as well as a long C-terminal cytoplasmic tail possessing a characteristic 25-amino-acid sequence. Rag4 may therefore be expected to have a glucose-sensing function. However, the rag4 mutation was fully complemented by one copy of either SNF3 or RGT2. Since K. lactis appears to have no other genes of the SNF3/RGT2 type, we suggest that Rag4 of K. lactis may have a dual function of signaling high and low concentrations of glucose. In rag4 mutants, glucose repression of several inducible enzymes is abolished. PMID- 11404321 TI - A role for the Swe1 checkpoint kinase during filamentous growth of Saccharomyces cerevisiae. AB - In this study we show that inactivation of Hsl1 or Hsl7, negative regulators of the Swe1 kinase, enhances the invasive behavior of haploid and diploid cells. The enhancement of filamentous growth caused by inactivation of both genes is mediated via the Swe1 protein kinase. Whereas Swe1 contributes noticeably to the effectiveness of haploid invasive growth under all conditions tested, its contribution to pseudohyphal growth is limited to the morphological response under standard assay conditions. However, Swe1 is essential for pseudohyphal differentiation under a number of nonstandard assay conditions including altered temperature and increased nitrogen. Swe1 is also required for pseudohyphal growth in the absence of Tec1 and for the induction of filamentation by butanol, a related phenomenon. Although inactivation of Hsl1 is sufficient to suppress the defect in filamentous growth caused by inactivation of Tec1 or Flo8, it is insufficient to promote filamentous growth in the absence of both factors. Moreover, inactivation of Hsl1 will not bypass the requirement for nitrogen starvation or growth on solid medium for pseudohyphal differentiation. We conclude that the Swe1 kinase modulates filamentous development under a broad spectrum of conditions and that its role is partially redundant with the Tec1 and Flo8 transcription factors. PMID- 11404322 TI - Interaction of the repressors Nrg1 and Nrg2 with the Snf1 protein kinase in Saccharomyces cerevisiae. AB - The Snf1 protein kinase is essential for the transcription of glucose-repressed genes in Saccharomyces cerevisiae. We identified Nrg2 as a protein that interacts with Snf1 in the two-hybrid system. Nrg2 is a C(2)H(2) zinc-finger protein that is homologous to Nrg1, a repressor of the glucose- and Snf1-regulated STA1 (glucoamylase) gene. Snf1 also interacts with Nrg1 in the two-hybrid system and co-immunoprecipitates with both Nrg1 and Nrg2 from cell extracts. A LexA fusion to Nrg2 represses transcription from a promoter containing LexA binding sites, indicating that Nrg2 also functions as a repressor. An Nrg1 fusion to green fluorescent protein is localized to the nucleus, and this localization is not regulated by carbon source. Finally, we show that VP16 fusions to Nrg1 and Nrg2 allow low-level expression of SUC2 in glucose-grown cells, and we present evidence that Nrg1 and Nrg2 contribute to glucose repression of the DOG2 gene. These results suggest that Nrg1 and Nrg2 are direct or indirect targets of the Snf1 kinase and function in glucose repression of a subset of Snf1-regulated genes. PMID- 11404323 TI - Rpm2, the protein subunit of mitochondrial RNase P in Saccharomyces cerevisiae, also has a role in the translation of mitochondrially encoded subunits of cytochrome c oxidase. AB - RPM2 is a Saccharomyces cerevisiae nuclear gene that encodes the protein subunit of mitochondrial RNase P and has an unknown function essential for fermentative growth. Cells lacking mitochondrial RNase P cannot respire and accumulate lesions in their mitochondrial DNA. The effects of a new RPM2 allele, rpm2-100, reveal a novel function of RPM2 in mitochondrial biogenesis. Cells with rpm2-100 as their only source of Rpm2p have correctly processed mitochondrial tRNAs but are still respiratory deficient. Mitochondrial mRNA and rRNA levels are reduced in rpm2-100 cells compared to wild type. The general reduction in mRNA is not reflected in a similar reduction in mitochondrial protein synthesis. Incorporation of labeled precursors into mitochondrially encoded Atp6, Atp8, Atp9, and Cytb protein was enhanced in the mutant relative to wild type, while incorporation into Cox1p, Cox2p, Cox3p, and Var1p was reduced. Pulse-chase analysis of mitochondrial translation revealed decreased rates of translation of COX1, COX2, and COX3 mRNAs. This decrease leads to low steady-state levels of Cox1p, Cox2p, and Cox3p, loss of visible spectra of aa(3) cytochromes, and low cytochrome c oxidase activity in mutant mitochondria. Thus, RPM2 has a previously unrecognized role in mitochondrial biogenesis, in addition to its role as a subunit of mitochondrial RNase P. Moreover, there is a synthetic lethal interaction between the disruption of this novel respiratory function and the loss of wild-type mtDNA. This synthetic interaction explains why a complete deletion of RPM2 is lethal. PMID- 11404324 TI - Yeast ASF1 protein is required for cell cycle regulation of histone gene transcription. AB - Transcription of the four yeast histone gene pairs (HTA1-HTB1, HTA2-HTB2, HHT1 HHF1, and HHT2-HHF2) is repressed during G1, G2, and M. For all except HTA2-HTB2, this repression requires several trans-acting factors, including the products of the HIR genes, HIR1, HIR2, and HIR3. ASF1 is a highly conserved protein that has been implicated in transcriptional silencing and chromatin assembly. In this analysis, we show that HIR1 interacts with ASF1 in a two-hybrid analysis. Further, asf1 mutants, like hir mutants, are defective in repression of histone gene transcription during the cell cycle and in cells arrested in early S phase in response to hydroxyurea. asf1 and hir1 mutations also show very similar synergistic interactions with mutations in cac2, a subunit of the yeast chromatin assembly factor CAF-I. The results suggest that ASF1 and HIR1 function in the same pathway to create a repressive chromatin structure in the histone genes during the cell cycle. PMID- 11404325 TI - Yeast spt6-140 mutation, affecting chromatin and transcription, preferentially increases recombination in which Rad51p-mediated strand exchange is dispensable. AB - We have shown that the spt6-140 and spt4-3 mutations, affecting chromatin structure and transcription, stimulate recombination between inverted repeats by a RAD52-dependent mechanism that is very efficient in the absence of RAD51, RAD54, RAD55, and RAD57. Such a mechanism of recombination is RAD1-RAD59 dependent and yields gene conversions highly associated with the inversion of the repeat. The spt6-140 mutation alters transcription and chromatin in our inverted repeats, as determined by Northern and micrococcal nuclease sensitivity analyses, respectively. Hyper-recombination levels are diminished in the absence of transcription. We believe that the chromatin alteration, together with transcription impairment caused by spt6-140, increases the incidence of spontaneous recombination regardless of whether or not it is mediated by Rad51p dependent strand exchange. Our results suggest that spt6, as well as spt4, primarily stimulates a mechanism of break-induced replication. We discuss the possibility that the chromatin alteration caused by spt6-140 facilitates a Rad52p mediated one-ended strand invasion event, possibly inefficient in wild-type chromatin. Our results are consistent with the idea that the major mechanism leading to inversions might not be crossing over but break-induced replication followed by single-strand annealing. PMID- 11404326 TI - The Saccharomyces cerevisiae small GTPase, Gsp1p/Ran, is involved in 3' processing of 7S-to-5.8S rRNA and in degradation of the excised 5'-A0 fragment of 35S pre-rRNA, both of which are carried out by the exosome. AB - Dis3p, a subunit of the exosome, interacts directly with Ran. To clarify the relationship between the exosome and the RanGTPase cycle, a series of temperature sensitive Saccharomyces cerevisiae dis3 mutants were isolated and their 5.8S rRNA processing was compared with processing in strains with mutations in a S. cerevisiae Ran homologue, Gsp1p. In both dis3 and gsp1 mutants, 3' processing of 7S-to-5.8S rRNA was blocked at three identical sites in an allele-specific manner. In contrast, the 5' end of 5.8S rRNA was terminated normally in gsp1 and in dis3. Inhibition of 5.8S rRNA maturation in gsp1 was rescued by overexpression of nuclear exosome components Dis3p, Rrp4p, and Mtr4p, but not by a cytoplasmic exosome component, Ski2p. Furthermore, gsp1 and dis3 accumulated the 5'-A0 fragment of 35S pre-rRNA, which is also degraded by the exosome, and the level of 27S rRNA was reduced. Neither 5.8S rRNA intermediates nor 5'-A0 fragments were observed in mutants defective in the nucleocytoplasmic transport, indicating that Gsp1p regulates rRNA processing through Dis3p, independent of nucleocytoplasmic transport. PMID- 11404327 TI - Genetic evidence supports a role for the yeast CCR4-NOT complex in transcriptional elongation. AB - The CCR4-NOT complex is involved in the regulation of gene expression both positively and negatively. The repressive effects of the complex appear to result in part from restricting TBP access to noncanonical TATAA binding sites presumably through interaction with multiple TAF proteins. We provide here genetic evidence that the CCR4-NOT complex also plays a role in transcriptional elongation. First, defects in CCR4-NOT components as well as overexpression of the NOT4 gene elicited 6-azauracil (6AU) and mycophenolic acid sensitivities, hallmarks of transcriptional elongation defects. A number of other transcription initiation factors known to interact with the CCR4-NOT complex did not elicit these phenotypes nor did defects in factors that reduced mRNA degradation and hence the recycling of NTPs. Second, deletion of ccr4 resulted in severe synthetic effects with mutations or deletions in the known elongation factors RPB2, TFIIS, and SPT16. Third, the ccr4 deletion displayed allele-specific interactions with rpb1 alleles that are thought to be important in the control of elongation. Finally, we found that a ccr4 deletion as well as overexpression of the NOT1 gene specifically suppressed the cold-sensitive phenotype associated with the spt5-242 allele. The only other known suppressors of this spt5-242 allele are factors involved in slowing transcriptional elongation. These genetic results are consistent with the model that the CCR4-NOT complex, in addition to its known effects on initiation, plays a role in aiding the elongation process. PMID- 11404328 TI - Intersexual partial diploids of phycomyces. AB - Sexual interaction between strains of opposite sex in many fungi of the order Mucorales modifies hyphal morphology and increases the carotene content. The progeny of crosses of Phycomyces blakesleeanus usually include a small proportion of anomalous segregants that show these signs of sexual stimulation without a partner. We have analyzed the genetic constitution of such segregants from crosses that involved a carF mutation for overaccumulation of beta-carotene and other markers. The new strains were diploids or partial diploids heterozygous for the sex markers. Diploidy was unknown in this fungus and in the Zygomycetes. Random chromosome losses during the vegetative growth of the diploid led to heterokaryosis in the coenocytic mycelia and eventually to sectors of various tints and mating behavior. The changes in the nuclear composition of the mycelia could be followed by selecting for individual nuclei. The results impose a reinterpretation of the sexual cycle of Phycomyces. Some of the intersexual strains that carried the carF mutation contained 25 mg beta-carotene per gram of dry mass and were sufficiently stable for practical use in carotene production. PMID- 11404329 TI - Goalpha regulates volatile anesthetic action in Caenorhabditis elegans. AB - To identify genes controlling volatile anesthetic (VA) action, we have screened through existing Caenorhabditis elegans mutants and found that strains with a reduction in Go signaling are VA resistant. Loss-of-function mutants of the gene goa-1, which codes for the alpha-subunit of Go, have EC(50)s for the VA isoflurane of 1.7- to 2.4-fold that of wild type. Strains overexpressing egl-10, which codes for an RGS protein negatively regulating goa-1, are also isoflurane resistant. However, sensitivity to halothane, a structurally distinct VA, is differentially affected by Go pathway mutants. The RGS overexpressing strains, a goa-1 missense mutant found to carry a novel mutation near the GTP-binding domain, and eat-16(rf) mutants, which suppress goa-1(gf) mutations, are all halothane resistant; goa-1(null) mutants have wild-type sensitivities. Double mutant strains carrying mutations in both goa-1 and unc-64, which codes for a neuronal syntaxin previously found to regulate VA sensitivity, show that the syntaxin mutant phenotypes depend in part on goa-1 expression. Pharmacological assays using the cholinesterase inhibitor aldicarb suggest that VAs and GOA-1 similarly downregulate cholinergic neurotransmitter release in C. elegans. Thus, the mechanism of action of VAs in C. elegans is regulated by Goalpha, and presynaptic Goalpha-effectors are candidate VA molecular targets. PMID- 11404330 TI - Regions of lower crossing over harbor more rare variants in African populations of Drosophila melanogaster. AB - A correlation between diversity levels and rates of recombination is predicted both by models of positive selection, such as hitchhiking associated with the rapid fixation of advantageous mutations, and by models of purifying selection against strongly deleterious mutations (commonly referred to as "background selection"). With parameter values appropriate for Drosophila populations, only the first class of models predicts a marked skew in the frequency spectrum of linked neutral variants, relative to a neutral model. Here, we consider 29 loci scattered throughout the Drosophila melanogaster genome. We show that, in African populations, a summary of the frequency spectrum of polymorphic mutations is positively correlated with the meiotic rate of crossing over. This pattern is demonstrated to be unlikely under a model of background selection. Models of weakly deleterious selection are not expected to produce both the observed correlation and the extent to which nucleotide diversity is reduced in regions of low (but nonzero) recombination. Thus, of existing models, hitchhiking due to the recurrent fixation of advantageous variants is the most plausible explanation for the data. PMID- 11404331 TI - A systematic screen for dominant second-site modifiers of Merlin/NF2 phenotypes reveals an interaction with blistered/DSRF and scribbler. AB - Merlin, the Drosophila homologue of the human tumor suppressor gene Neurofibromatosis 2 (NF2), is required for the regulation of cell proliferation and differentiation. To better understand the cellular functions of the NF2 gene product, Merlin, recent work has concentrated on identifying proteins with which it interacts either physically or functionally. In this article, we describe genetic screens designed to isolate second-site modifiers of Merlin phenotypes from which we have identified five multiallelic complementation groups that modify both loss-of-function and dominant-negative Merlin phenotypes. Three of these groups, Group IIa/scribbler (also known as brakeless), Group IIc/blistered, and Group IId/net, are known genes, while two appear to be novel. In addition, two genes, Group IIa/scribbler and Group IIc/blistered, alter Merlin subcellular localization in epithelial and neuronal tissues, suggesting that they regulate Merlin trafficking or function. Furthermore, we show that mutations in scribbler and blistered display second-site noncomplementation with one another. These results suggest that Merlin, blistered, and scribbler function together in a common pathway to regulate Drosophila wing epithelial development. PMID- 11404332 TI - The rate of mutation and the homozygous and heterozygous mutational effects for competitive viability: a long-term experiment with Drosophila melanogaster. AB - The effect of 250 generations of mutation accumulation (MA) on the second chromosome competitive viability of Drosophila melanogaster was analyzed both in homozygous and heterozygous conditions. We used full-sib MA lines, where selection hampers the accumulation of severely deleterious mutations but is ineffective against mildly deleterious ones. A large control population was simultaneously evaluated. Competitive viability scores, unaffected by the expression of mutations in heterozygosis, were obtained relative to a Cy/L(2) genotype. The rate of decline in mean DeltaM approximately 0.1% was small. However, that of increase in variance DeltaV approximately 0.08 x 10(-3) was similar to the values obtained in previous experiments when severely deleterious mutations were excluded. The corresponding estimates of the mutation rate lambda > or = 0.01 and the average effect of mutations E(s) < or = 0.08 are in good agreement with Bateman-Mukai and minimum distance estimates for noncompetitive viability obtained from the same MA lines after 105 generations. Thus, competitive and noncompetitive viability show similar mutational properties. The regression estimate of the degree of dominance for mild-to-moderate deleterious mutations was approximately 0.3, suggesting that the pertinent value for new unselected mutations should be somewhat smaller. PMID- 11404333 TI - AFLP-based genetic linkage map of the Colorado potato beetle Leptinotarsa decemlineata: sex chromosomes and a pyrethroid-resistance candidate gene. AB - A genetic linkage map was constructed from an intraspecific cross of the Colorado potato beetle, Leptinotarsa decemlineata. This is an initial step toward mapping the loci that underlie important phenotypes associated with insect adaptation to an agroecosystem. The map was made with 172 AFLP and 10 anonymous codominant markers segregating among 74 backcross (BC(1)) individuals. Markers were mapped to 18 linkage groups and a subset of the markers with a mean intermarker distance of 11.1 cM is presented. A pyrethroid-resistance candidate gene, LdVssc1, was placed onto the map as well. The sex chromosome was identified by exploiting the XO nature of sex determination in this species using patterns of variation at LdVssc1 and the codominant markers. PMID- 11404334 TI - Characterization of the flamenco region of the Drosophila melanogaster genome. AB - The flamenco gene, located at 20A1-3 in the beta-heterochromatin of the Drosophila X chromosome, is a major regulator of the gypsy/mdg4 endogenous retrovirus. As a first step to characterize this gene, approximately 100 kb of genomic DNA flanking a P-element-induced mutation of flamenco was isolated. This DNA is located in a sequencing gap of the Celera Genomics project, i.e., one of those parts of the genome in which the "shotgun" sequence could not be assembled, probably because it contains long stretches of repetitive DNA, especially on the proximal side of the P insertion point. Deficiency mapping indicated that sequences required for the normal flamenco function are located >130 kb proximal to the insertion site. The distal part of the cloned DNA does, nevertheless, contain several unique sequences, including at least four different transcription units. Dip1, the closest one to the P-element insertion point, might be a good candidate for a gypsy regulator, since it putatively encodes a nuclear protein containing two double-stranded RNA-binding domains. However, transgenes containing dip1 genomic DNA were not able to rescue flamenco mutant flies. The possible nature of the missing flamenco sequences is discussed. PMID- 11404335 TI - SSCP analysis of cDNA markers provides a dense linkage map of the Aedes aegypti genome. AB - An intensive linkage map of the yellow fever mosquito, Aedes aegypti, was constructed using single-strand conformation polymorphism (SSCP) analysis of cDNA markers to identify single nucleotide polymorphisms (SNPs). A total of 94 A. aegypti cDNAs were downloaded from GenBank and primers were designed to amplify fragments <500 bp in size. These primer pairs amplified 94 loci, 57 (61%) of which segregated in a single F(1) intercross family among 83 F(2) progeny. This allowed us to produce a dense linkage map of one marker every 2 cM distributed over a total length of 134 cM. Many A. aegypti cDNAs were highly similar to genes in the Drosophila melanogaster genome project. Comparative linkage analysis revealed areas of synteny between the two species. SNP polymorphisms are abundant in A. aegypti genes and should prove useful in both population genetics and mapping studies. PMID- 11404336 TI - A microsatellite-based genetic linkage map for channel catfish, Ictalurus punctatus. AB - Microsatellite loci were identified in channel catfish gene sequences or random clones from a small insert genomic DNA library. Outbred populations of channel catfish contained an average of eight alleles per locus and an average heterozygosity of 0.70. A genetic linkage map of the channel catfish genome (N = 29) was constructed from two reference families. A total of 293 microsatellite loci were polymorphic in one or both families, with an average of 171 informative meioses per locus. Nineteen type I loci, 243 type II loci, and one EST were placed in 32 multipoint linkage groups covering 1958 cM. Nine more type II loci were contained in three two-point linkage groups covering 24.5 cM. Twenty-two type II loci remained unlinked. Multipoint linkage groups ranged in size from 11.9 to 110.5 cM with an average intermarker distance of 8.7 cM. Seven microsatellite loci were closely linked with the sex-determining locus. The microsatellite loci and genetic linkage map will increase the efficiency of selective breeding programs for channel catfish. PMID- 11404337 TI - Conserved vertebrate chromosome segments in the large salamander genome. AB - Urodele amphibians (salamanders) are important models for embryological, physiological, and natural history research and are also a biomedically important group because they are the only vertebrates capable of regenerating entire organ systems. To enhance the utility of salamanders for biomedical research and for understanding genome evolution, genetic linkage analysis was used to identify chromosome segments that are homologous between ambystomatid salamanders and distantly related vertebrate model organisms. A total of 347 loci (AFLPs, RAPDs, and protein-coding loci) were mapped using an interspecific meiotic mapping panel (Ambystoma mexicanum and A. tigrinum tigrinum; family Ambystomatidae). Genome size in Ambystoma was estimated to be 7291 cM, the largest linkage map estimate reported for any organism. However, the relatively large size of the salamander genome did not hinder efforts to map and identify conserved syntenies from a small sample of 24 protein-coding loci. Chromosomal segments that are conserved between fishes and mammals are also conserved in these salamanders. Thus, comparative gene mapping appears to be an efficient strategy for identifying orthologous loci between ambystomatid salamanders and genomically well characterized vertebrate model organisms. PMID- 11404338 TI - Contrasting patterns of polymorphisms at the ABO-secretor gene (FUT2) and plasma alpha(1,3)fucosyltransferase gene (FUT6) in human populations. AB - The coding sequences ( approximately 1 kb) of FUT2 [ABO-Secretor type alpha(1,2)fucosyltransferase] and of FUT6 [plasma alpha(1,3)fucosyltransferase] were analyzed for allelic polymorphism by direct sequencing in five populations. The nucleotide diversities of FUT2 estimated from pairwise sequence differences were 0.0045, 0.0042, 0.0042, 0.0009, and 0.0008 in Africans, European-Africans, Iranians, Chinese, and Japanese, respectively. The nucleotide diversities of FUT6 were 0.0024, 0.0016, 0.0015, 0.0017, and 0.0020 in Africans, European-Africans, Iranians, Chinese, and Japanese, respectively. At FUT2, excesses in pairwise sequence differences compared to the number of polymorphic sites as indicated by a significantly positive Tajima's D were observed in European-Africans and in Iranians. The data do not fit expectations of the equilibrium neutral model with an infinite number of sites. On the other hand, Tajima's D's at FUT6 in each of the five populations and at FUT2 in Africans, Chinese, and Japanese were not significantly different from zero. F(ST) between the Asians and the others measured at FUT2 was higher than at FUT6. These results suggest that natural selection was responsible for the generation of the FUT2 polymorphism in European Africans and in Iranians. PMID- 11404339 TI - Interchromosomal gene conversion at an endogenous human cell locus. AB - To examine the relationship between gene conversion and reciprocal exchange at an endogenous chromosomal locus, we developed a reversion assay in a thymidine kinase deficient mutant, TX545, derived from the human lymphoblastoid cell line TK6. Selectable revertants of TX545 can be generated through interchromosomal gene conversion at the site of inactivating mutations on each tk allele or by reciprocal exchange that alters the linkage relationships of inactivating polymorphisms within the tk locus. Analysis of loss of heterozygosity (LOH) at intragenic polymorphisms and flanking microsatellite markers was used to initially evaluate allelotypes in TK(+) revertants for patterns associated with either gene conversion or crossing over. The linkage pattern in a subset of convertants was then unambiguously established, even in the event of prereplicative recombinational exchanges, by haplotype analysis of flanking microsatellite loci in tk(-/-) LOH mutants collected from the tk(+/-) parental convertant. Some (7/38; 18%) revertants were attributable to easily discriminated nonrecombinational mechanisms, including suppressor mutations within the tk coding sequence. However, all revertants classified as a recombinational event (28/38; 74%) were attributed to localized gene conversion, representing a highly significant preference (P < 0.0001) over gene conversion with associated reciprocal exchange, which was never observed. PMID- 11404340 TI - An ancient retrovirus-like element contains hot spots for SINE insertion. AB - Vertebrate retrotransposons have been used extensively for phylogenetic analyses and studies of molecular evolution. Information can be obtained from specific inserts either by comparing sequence differences that have accumulated over time in orthologous copies of that insert or by determining the presence or absence of that specific element at a particular site. The presence of specific copies has been deemed to be an essentially homoplasy-free phylogenetic character because the probability of multiple independent insertions into any one site has been believed to be nil. Mys elements are a type of LTR-containing retrotransposon present in Sigmodontine rodents. In this study we have shown that one particular insert, mys-9, is an extremely old insert present in multiple species of the genus Peromyscus. We have found that different copies of this insert show a surprising range of sizes, due primarily to a continuing series of SINE (short interspersed element) insertions into this locus. We have identified two hot spots for SINE insertion within mys-9 and at each hot spot have found that two independent SINE insertions have occurred at identical sites. These results have major repercussions for phylogenetic analyses based on SINE insertions, indicating the need for caution when one concludes that the existence of a SINE at a specific locus in multiple individuals is indicative of common ancestry. Although independent insertions at the same locus may be rare, SINE insertions are not homoplasy-free phylogenetic markers. PMID- 11404341 TI - A frameshift mutation in MC1R and a high frequency of somatic reversions cause black spotting in pigs. AB - Black spotting on a red or white background in pigs is determined by the E(P) allele at the MC1R/Extension locus. A previous comparison of partial MC1R sequences revealed that E(P) shares a missense mutation (D121N) with the E(D2) allele for dominant black color. Sequence analysis of the entire coding region now reveals a second mutation in the form of a 2-bp insertion at codon 23 (nt67insCC). This mutation expands a tract of six C nucleotides to eight and introduces a premature stop codon at position 56. This frameshift mutation is expected to cause a recessive red color, which was in fact observed in some breeds with the E(P) allele present (Tamworth and Hereford). RT-PCR analyses were conducted using skin samples taken from both spotted and background areas of spotted pigs. The background red area had transcript only from the mutant nt67insCC MC1R allele, whereas the black spot also contained a transcript without the 2-bp insertion. This indicates that black spots are due to somatic reversion events that restore the frame and MC1R function. The phenotypic expression of the E(P) allele is highly variable and the associated coat color ranges from red, red with black spots, white with black spots, to almost completely solid black. In several breeds of pigs the phenotypic manifestation of this allele has been modified by selection for or against black spots. PMID- 11404342 TI - Dense genetic linkage maps of three Populus species (Populus deltoides, P. nigra and P. trichocarpa) based on AFLP and microsatellite markers. AB - Populus deltoides, P. nigra, and P. trichocarpa are the most important species for poplar breeding programs worldwide. In addition, Populus has become a model for fundamental research on trees. Linkage maps were constructed for these three species by analyzing progeny of two controlled crosses sharing the same female parent, Populus deltoides cv. S9-2 x P. nigra cv. Ghoy and P. deltoides cv. S9-2 x P. trichocarpa cv. V24. The two-way pseudotestcross mapping strategy was used to construct the maps. Amplified fragment length polymorphism (AFLP) markers that segregated 1:1 were used to form the four parental maps. Microsatellites and sequence-tagged sites were used to align homoeologous groups between the maps and to merge linkage groups within the individual maps. Linkage analysis and alignment of the homoeologous groups resulted in 566 markers distributed over 19 groups for P. deltoides covering 86% of the genome, 339 markers distributed over 19 groups for P. trichocarpa covering 73%, and 369 markers distributed over 28 groups for P. nigra covering 61%. Several tests for randomness showed that the AFLP markers were randomly distributed over the genome. PMID- 11404343 TI - Genomic organization of plant terpene synthases and molecular evolutionary implications. AB - Terpenoids are the largest, most diverse class of plant natural products and they play numerous functional roles in primary metabolism and in ecological interactions. The first committed step in the formation of the various terpenoid classes is the transformation of the prenyl diphosphate precursors, geranyl diphosphate, farnesyl diphosphate, and geranylgeranyl diphosphate, to the parent structures of each type catalyzed by the respective monoterpene (C(10)), sesquiterpene (C(15)), and diterpene synthases (C(20)). Over 30 cDNAs encoding plant terpenoid synthases involved in primary and secondary metabolism have been cloned and characterized. Here we describe the isolation and analysis of six genomic clones encoding terpene synthases of conifers, [(-)-pinene (C(10)), (-) limonene (C(10)), (E)-alpha-bisabolene (C(15)), delta-selinene (C(15)), and abietadiene synthase (C(20)) from Abies grandis and taxadiene synthase (C(20)) from Taxus brevifolia], all of which are involved in natural products biosynthesis. Genome organization (intron number, size, placement and phase, and exon size) of these gymnosperm terpene synthases was compared to eight previously characterized angiosperm terpene synthase genes and to six putative terpene synthase genomic sequences from Arabidopsis thaliana. Three distinct classes of terpene synthase genes were discerned, from which assumed patterns of sequential intron loss and the loss of an unusual internal sequence element suggest that the ancestral terpenoid synthase gene resembled a contemporary conifer diterpene synthase gene in containing at least 12 introns and 13 exons of conserved size. A model presented for the evolutionary history of plant terpene synthases suggests that this superfamily of genes responsible for natural products biosynthesis derived from terpene synthase genes involved in primary metabolism by duplication and divergence in structural and functional specialization. This novel molecular evolutionary approach focused on genes of secondary metabolism may have broad implications for the origins of natural products and for plant phylogenetics in general. PMID- 11404344 TI - A quantitative genetic analysis of nuclear-cytoplasmic male sterility in structured populations of Silene vulgaris. AB - Gynodioecy, the coexistence of functionally female and hermaphroditic morphs within plant populations, often has a complicated genetic basis involving several cytoplasmic male-sterility factors and nuclear restorers. This complexity has made it difficult to study the genetics and evolution of gynodioecy in natural populations. We use a quantitative genetic analysis of crosses within and among populations of Silene vulgaris to partition genetic variance for sex expression into nuclear and cytoplasmic components. We also use mitochondrial markers to determine whether cytoplasmic effects on sex expression can be traced to mitochondrial variance. Cytoplasmic variation and epistatic interactions between nuclear and cytoplasmic loci accounted for a significant portion of the variation in sex expression among the crosses. Source population also accounted for a significant portion of the sex ratio variation. Crosses among populations greatly enhanced the dam (cytoplasmic) effect, indicating that most among-population variance was at cytoplasmic loci. This is supported by the large among-population variance in the frequency of mitochondrial haplotypes, which also accounted for a significant portion of the sex ratio variance in our data. We discuss the similarities between the population structure we observed at loci that influence sex expression and previous work on putatively neutral loci, as well as the implications this has for what mechanisms may create and maintain population structure at loci that are influenced by natural selection. PMID- 11404345 TI - Cytonuclear disequilibrium and genetic drift in a natural population of ponderosa pine. AB - We measured the cytonuclear disequilibrium between 11 nuclear allozyme loci and both mitochondrial and chloroplast DNA haplotypes in a natural population of ponderosa pine (Pinus ponderosa, Laws). Three allozyme loci showed significant associations with mtDNA variation, while two other loci showed significant association with cpDNA. However, the absolute number of individuals involved in any of the associations was small, such that in none of the nuclear-organellar combinations was the difference between observed and expected numbers >11 individuals. Patterns of association were not consistent across loci or organellar genomes, suggesting that they are not the result of mating patterns, which would act uniformly on all loci. This pattern of disequilibria is consistent with the action of genetic drift and with existing knowledge of the structure of this population and thus does not imply the action of other evolutionary processes. The overall magnitude (normalized disequilibrium) of associations was greater for maternally inherited mtDNA than for paternally inherited cpDNA, though this difference was neither large nor significant. Such significant disequilibria involving the paternally inherited organelle indicate that not only are there a limited number of seed parents, but the effective number of pollen parents is also limited. PMID- 11404346 TI - Tracing evolutionary and developmental implications of mitochondrial stoichiometric shifting in the common bean. AB - The recombination and copy number shifting activities of the plant mitochondrial genome are widely documented across plant genera, but these genome processes have not been as well examined with regard to their roles in plant evolution. Because of the extensive plant collections of Phaseolus spp and the degree to which cytoplasmic male sterility (cms) has been characterized in the common bean, this system would be valuable for investigating mitochondrial genome dynamics in natural populations. We have used the cms-associated sequence pvs-orf239 as a mitochondrial genetic marker for these studies and have demonstrated its universal presence throughout a diversity of undomesticated Phaseolus lines. Within these populations, the pvs-orf239 sequence is present in high copy number in approximately 10% of the lines, but substoichiometric in all others. This mitochondrial sequence, derived apparently by at least two recombination events, is well conserved with two point mutations identified that are both apparently silent with regard to the sterility phenotype. A putative progenitor sequence was identified in Phaseolus glabelus in substoichiometric levels, suggesting that the present-day pvs-orf239 sequence was likely introduced substoichiometrically. Copy number shifting within the mitochondrial genome results in a 1000- to 2000-fold change, so that substoichiometric forms are estimated at less than one copy per every 100 cells. On the basis of PCR analysis of root tips, we postulate that a mitochondrial "transmitted form" resides within the meristem to assure transmission of a complete genetic complement to progeny. PMID- 11404347 TI - The use of intraallelic variability for testing neutrality and estimating population growth rate. AB - To better understand the forces affecting individual alleles, we introduce a method for finding the joint distribution of the frequency of a neutral allele and the extent of variability at closely linked marker loci (the intraallelic variability). We model three types of intraallelic variability: (a) the number of nonrecombinants at a linked biallelic marker locus, (b) the length of a conserved haplotype, and (c) the number of mutations at a linked marker locus. If the population growth rate is known, the joint distribution provides the basis for a test of neutrality by testing whether the observed level of intraallelic variability is consistent with the observed allele frequency. If the population growth rate is unknown but neutrality can be assumed, the joint distribution provides the likelihood of the growth rate and leads to a maximum-likelihood estimate. We apply the method to data from published data sets for four loci in humans. We conclude that the Delta32 allele at CCR5 and a disease-associated allele at MLH1 arose recently and have been subject to strong selection. Alleles at PAH appear to be neutral and we estimate the recent growth rate of the European population to be approximately 0.027 per generation with a support interval of (0.017-0.037). Four of the relatively common alleles at CFTR also appear to be neutral but DeltaF508 appears to be significantly advantageous to heterozygous carriers. PMID- 11404348 TI - An unconditional exact test for the Hardy-Weinberg equilibrium law: sample-space ordering using the Bayes factor. AB - Much forensic inference based upon DNA evidence is made assuming that the Hardy Weinberg equilibrium (HWE) is valid for the genetic loci being used. Several statistical tests to detect and measure deviation from HWE have been devised, each having advantages and limitations. The limitations become more obvious when testing for deviation within multiallelic DNA loci is attempted. Here we present an exact test for HWE in the biallelic case, based on the ratio of weighted likelihoods under the null and alternative hypotheses, the Bayes factor. This test does not depend on asymptotic results and minimizes a linear combination of type I and type II errors. By ordering the sample space using the Bayes factor, we also define a significance (evidence) index, P value, using the weighted likelihood under the null hypothesis. We compare it to the conditional exact test for the case of sample size n = 10. Using the idea under the method of chi(2) partition, the test is used sequentially to test equilibrium in the multiple allele case and then applied to two short tandem repeat loci, using a real Caucasian data bank, showing its usefulness. PMID- 11404349 TI - Distinguishing migration from isolation: a Markov chain Monte Carlo approach. AB - A Markov chain Monte Carlo method for estimating the relative effects of migration and isolation on genetic diversity in a pair of populations from DNA sequence data is developed and tested using simulations. The two populations are assumed to be descended from a panmictic ancestral population at some time in the past and may (or may not) after that be connected by migration. The use of a Markov chain Monte Carlo method allows the joint estimation of multiple demographic parameters in either a Bayesian or a likelihood framework. The parameters estimated include the migration rate for each population, the time since the two populations diverged from a common ancestral population, and the relative size of each of the two current populations and of the common ancestral population. The results show that even a single nonrecombining genetic locus can provide substantial power to test the hypothesis of no ongoing migration and/or to test models of symmetric migration between the two populations. The use of the method is illustrated in an application to mitochondrial DNA sequence data from a fish species: the threespine stickleback (Gasterosteus aculeatus). PMID- 11404350 TI - Estimating the time to the most recent common ancestor for the Y chromosome or mitochondrial DNA for a pair of individuals. AB - Bayesian posterior distributions are obtained for the time to the most recent common ancestor (MRCA) for a nonrecombining segment of DNA (such as the nonpseudoautosomal arm of the Y chromosome or the mitochondrial genome) for two individuals given that they match at k out of n scored markers. We argue that the distribution of the time t to the MRCA is the most natural measure of relatedness for such nonrecombining regions. Both an infinite-alleles (no recurring mutants) and stepwise mutation model are examined, and these agree well when n is moderate to large and k/n is close to one. As expected, the infinite alleles model underestimates t relative to the stepwise model. Using a modest number (20) of microsatellite markers is sufficient to obtain reasonably precise estimates of t for individuals separated by 200 or less generations. Hence, the multilocus haplotypes of two individuals can be used not only to date very deep ancestry but also rather recent ancestry as well. Finally, our results have forensic implications in that a complete match at all markers between a suspect and a sample excludes only a modest subset of the population unless a very large number of markers (>500 microsatellites) are used. PMID- 11404351 TI - Recessive mutations and the maintenance of sex in structured populations. AB - The evolutionary maintenance of sexual reproduction remains a controversial problem. It was recently shown that recessive deleterious mutations create differences in the mutation load of sexual vs. asexual populations. Here we show that low levels of population structure or inbreeding can greatly enhance the importance of recessive deleterious mutations in the context of sexual vs. asexual populations. With population structure, the cost of sex can be substantially reduced or even eliminated for realistic levels of dominance. PMID- 11404352 TI - A histone deacetylation inhibitor and mutant promote colony-type switching of the human pathogen Candida albicans. AB - Most strains of Candida albicans undergo high frequency phenotypic switching. Strain WO-1 undergoes the white-opaque transition, which involves changes in colony and cellular morphology, gene expression, and virulence. We have hypothesized that the switch event involves heritable changes in chromatin structure. To test this hypothesis, we transiently exposed cells to the histone deacetylase inhibitor trichostatin-A (TSA). Treatment promoted a dramatic increase in the frequency of switching from white to opaque, but not opaque to white. Targeted deletion of HDA1, which encodes a deacetylase sensitive to TSA, had the same selective effect. These results support the model that the acetylation of histones plays a selective role in regulating the switching process. PMID- 11404353 TI - Alterations in the sensing and transport of phosphate and calcium by differentiating chondrocytes. AB - During endochondral bone formation and fracture healing, cells committed to chondrogenesis undergo a temporally restricted program of differentiation that is characterized by sequential changes in their phenotype and gene expression. This results in the manufacture, remodeling, and mineralization of a cartilage template on which bone is laid down. Articular chondrocytes undergo a similar but restricted differentiation program that does not proceed to mineralization, except in pathologic conditions such as osteoarthritis. The pathogenesis of disorders of cartilage development and metabolism, including osteochondrodysplasia, fracture non-union, and osteoarthritis remain poorly defined. We used the CFK2 model to examine the potential roles of phosphate and calcium ions in the regulatory pathways that mediate chondrogenesis and cartilage maturation. Differentiation was monitored over a 4-week period using a combination of morphological, biochemical, and molecular markers that have been characterized in vivo and in vitro. CFK2 cells expressed the type III sodium dependent phosphate transporters Glvr-1 and Ram-1, as well as a calcium-sensing mechanism. Regulated expression and activity of Glvr-1 by extracellular phosphate and parathyroid hormone-related protein was restricted to an early stage of CFK2 differentiation, as evidenced by expression of type II collagen, proteoglycan, and Ihh. On the other hand, regulated expression and activity of a calcium sensing receptor by extracellular calcium was most evident after 2 weeks of differentiation, concomitant with an increase in type X collagen expression, alkaline phosphatase activity and parathyroid hormone/parathyroid hormone-related protein receptor expression. On the basis of these temporally restricted changes in the sensing and transport of phosphate and calcium, we predict that extracellular phosphate plays a role in the commitment of chondrogenic cells to differentiation, whereas extracellular calcium plays a role at a later stage in their differentiation program. PMID- 11404354 TI - Distinction between nitrosating mechanisms within human cells and aqueous solution. AB - The quintessential nitrosating species produced during NO autoxidation is N(2)O(3). Nitrosation of amine, thiol, and hydroxyl residues can modulate critical cell functions. The biological mechanisms that control reactivity of nitrogen oxide species formed during autoxidation of nano- to micromolar levels of NO were examined using the synthetic donor NaEt(2)NN(O)NO (DEA/NO), human tumor cells, and 4,5-diaminofluorescein (DAF). Both the disappearance of NO and formation of nitrosated product from DAF in aerobic aqueous buffer followed second order processes; however, consumption of NO and nitrosation within intact cells were exponential. An optimal ratio of DEA/NO and 2-phenyl-4,4,5,5 tetramethylimidazole-1-oxyl 3-oxide (PTIO) was used to form N(2)O(3) through the intermediacy of NO(2). This route was found to be most reflective of the nitrosative mechanism within intact cells and was distinct from the process that occurred during autoxidation of NO in aqueous media. Manipulation of the endogenous scavengers ascorbate and glutathione indicated that the location, affinity, and concentration of these substances were key determinants in dictating nitrosative susceptibility of molecular targets. Taken together, these findings suggest that the functional effects of nitrosation may be organized to occur within discrete domains or compartments. Nitrosative stress may develop when scavengers are depleted and this architecture becomes compromised. Although NO(2) was not a component of aqueous NO autoxidation, the results suggest that the intermediacy of this species may be a significant factor in the advent of either nitrosation or oxidation chemistry in biological systems. PMID- 11404355 TI - Molecular characterization of the salutaridinol 7-O-acetyltransferase involved in morphine biosynthesis in opium poppy Papaver somniferum. AB - Salutaridinol 7-O-acetyltransferase (EC ) catalyzes the conversion of the phenanthrene alkaloid salutaridinol to salutaridinol-7-O-acetate, the immediate precursor of thebaine along the morphine biosynthetic pathway. We have isolated a cDNA clone that corresponds to the internal amino acid sequences of the native enzyme purified from a cell suspension culture of opium poppy Papaver somniferum. The recombinant enzyme acetylated the 7-hydroxyl moiety of salutaridinol in the presence of acetyl-CoA. The apparent K(m) value for salutaridinol was determined to be 9 microm and 54 microm for acetyl-CoA. The gene transcript was detected in extracts from Papaver orientale and Papaver bracteatum in addition to P. somniferum. Genomic DNA gel blot analysis indicated that there is likely a single copy of this gene in the P. somniferum genome. The amino acid sequence of salutaridinol 7-O-acetyltransferase is most similar (37% identity) to that of deacetylvindoline acetyltransferase of Catharanthus roseus. Salutaridinol 7-O acetyltransferase is the second enzyme specific to morphine biosynthesis for which we have isolated a cDNA. Taken together with the other cDNAs cloned encoding norcoclaurine 6-O-methyltransferase, (S)-N-methylcoclaurine 3' hydroxylase, the cytochrome P-450 reductase, and codeinone reductase, significant progress has been made toward accumulating genes of this pathway to enable the end goal of a biotechnological production of morphinan alkaloids. PMID- 11404356 TI - A conserved flagellar pocket exposed high mannose moiety is used by African trypanosomes as a host cytokine binding molecule. AB - Trypanosomes use antigenic variation of their variant-specific surface glycoprotein (VSG) coat as defense against the host immune system. However, in order to sustain their growth, they need to expose conserved epitopes, allowing host macromolecule binding and receptor-mediated endocytosis. Here we show that Trypanosoma brucei uses the conserved chitobiose-oligomannose (GlcNAc(2)-Man(5 9)) moieties of its VSG as a binding ligand for tumor necrosis factor (TNF), a host cytokine with lectin-like properties. As endocytosis in trypanosomes is restricted to the flagellar pocket, we show that soluble flagellar pocket extracts, and in particular soluble VSG, inhibit the binding of (125)I-TNF to trypanosomes. The interaction between TNF and VSG is confirmed by affinity chromatography, biosensor, and dot-blot affinity measurements, and soluble VSG inhibition of TNF-mediated trypanolysis. In all approaches, removal of N-linked carbohydrates abrogates the TNF-VSG interaction. In addition, synthetic high mannose oligosaccharides can block TNF-VSG interactions, and a VSG glycopeptide carrying the GlcNAc(2)-Man(5-9) moiety is shown to inhibit TNF-mediated trypanosome killing in mixed parasite/macrophage cell cultures. Together, these results support the observation that TNF plays a role in growth control of trypanosomes and, moreover, suggest that, by the use of conserved VSG carbohydrates as lectin-binding epitopes, trypanosomes can limit the necessity to express large numbers of invariant surface exposed receptors. PMID- 11404357 TI - Bid is cleaved by calpain to an active fragment in vitro and during myocardial ischemia/reperfusion. AB - Reperfusion after myocardial ischemia is associated with a rapid influx of calcium, leading to activation of various enzymes including calpain. Isolated perfused adult rabbit hearts subjected to global ischemia and reperfusion were studied. Calpain or a calpain-like activity was activated within 15 min after reperfusion, and preconditioning suppressed calpain activation. In contrast, caspase activation was not detected although cytochrome c was released after ischemia and reperfusion. The pro-apoptotic BH3-only Bcl-2 family member, Bid, was cleaved during ischemia/reperfusion in the adult rabbit heart. Recombinant Bid was cleaved by calpain to a fragment that was able to mediate cytochrome c release. The calpain cleavage site was mapped to a region within Bid that is extremely susceptible to proteolysis. These findings suggest that there is cross talk between apoptotic and necrotic pathways in myocardial ischemia/reperfusion injury. PMID- 11404358 TI - Stoichiometry, abundance, and functional significance of the hsp90/hsp70-based multiprotein chaperone machinery in reticulocyte lysate. AB - Rabbit reticulocyte lysate contains a multiprotein chaperone system that assembles the glucocorticoid receptor (GR) into a complex with hsp90 and converts the hormone binding domain of the receptor to its high affinity steroid binding state. This system has been resolved into five proteins, with hsp90 and hsp70 being essential and Hop, hsp40, and p23 acting as co-chaperones that optimize assembly. Hop binds independently to hsp70 and hsp90 to form an hsp90.Hop.hsp70 complex that acts as a machinery to open up the GR steroid binding site. Because purified hsp90 and hsp70 are sufficient for some activation of GR steroid binding activity, some investigators have rejected any role for Hop in GR.hsp90 heterocomplex assembly. Here, we counter that impression by showing that all of the Hop in reticulocyte lysate is present in an hsp90.Hop.hsp70 complex with a stoichiometry of 2:1:1. The complex accounts for approximately 30% of the hsp90 and approximately 9% of the hsp70 in lysate, and upon Sephacryl S-300 chromatography the GR.hsp90 assembly activity resides in the peak containing Hop bound hsp90. Consistent with the notion that the two essential chaperones cooperate with each other to open up the steroid binding site, we also show that purified hsp90 and hsp70 interact directly with each other to form weak hsp90.hsp70 complexes with a stoichiometry of 2:1. PMID- 11404359 TI - Identification of a missense mutation (G329A;Arg(110)--> GLN) in the human FUT7 gene. AB - The human FUT7 gene codes for the alpha1,3-fucosyltransferase VII (Fuc-TVII), which is involved in the biosynthesis of the sialyl Lewis x (SLe(x)) epitope on human leukocytes. The FUT7 gene has so far been considered to be monomorphic. Neutrophils isolated from patients with ulcerative colitis were examined for apparent alterations in protein glycosylation patterns by Western blot analysis using monoclonal antibodies directed against SLe(x) and SLe(x)-related epitopes. One individual showed lower levels of SLe(x) expression and an elevated expression of CD65s compared to controls. The coding regions of the FUT7 gene from this individual were cloned, and a G329A point mutation (Arg(110) --> Gln) was found in one allele, whereas the other FUT7 allele was wild type. No Fuc-TVII enzyme activity was detected in COS-7 cells transiently transfected with the mutated FUT7 construct. The FUT7 Arg(110) is conserved in all previously cloned vertebrate alpha 1,3-fucosyltransferases. Polymerase chain reaction followed by restriction enzyme cleavage was used to screen 364 unselected Caucasians for the G329A mutation, and a frequency of < or =1% for this mutation was found (3 heterozygotes). Genetic characterization of the family members of one of the additional heterozygotes identified one individual carrying the G329A mutation in both FUT7 alleles. Peripheral blood neutrophils of this homozygously mutated individual showed a lowered expression of SLe(x) and an elevated expression of CD65s when analyzed by Western blot and flow cytometry. The homozygous individual was diagnosed with ulcer disease, non-insulin-dependent diabetes, osteoporosis, spondyloarthrosis, and Sjogren's syndrome but had no history of recurrent bacterial infections or leukocytosis. PMID- 11404360 TI - Comparison of the Pseudomonas aeruginosa and Escherichia coli PhoQ sensor domains: evidence for distinct mechanisms of signal detection. AB - The PhoP-PhoQ two-component system is present in a number of Gram-negative bacteria where it has roles in Mg(2+) homeostasis and virulence. PhoQ is a transmembrane histidine kinase that activates PhoP-mediated regulation of a set of genes when the extracellular concentration of divalent cations is low. Divalent cations are thought to interact directly with the periplasmic PhoQ sensor domain. The PhoP-PhoQ systems of Escherichia coli and Pseudomonas aeruginosa are similar in their biological response to extracellular divalent cations; however, their sensor domains display little sequence identity. Here we have begun to explore the consequences of this sequence divergence by comparing the biophysical properties of the P. aeruginosa PhoQ sensor domain with the corresponding E. coli sensor domain. Unlike the E. coli protein, the P. aeruginosa PhoQ sensor domain undergoes changes in the circular dichroism and fluorescence spectra as well as destabilization of its dimeric form in response to divalent cations. These results suggest that distinct mechanisms of signal detection are utilized by these proteins. A hybrid protein in which the E. coli sensor domain has been substituted with the corresponding P. aeruginosa sensor domain responds normally to the presence of extracellular divalent cations in vivo in E. coli. Thus, despite apparent differences in the structural response to its stimulus, the P. aeruginosa sensor domain transduces signals to the E. coli PhoQ cytoplasmic kinase domain in a manner that mimics normal E. coli PhoQ function. PMID- 11404361 TI - chy1, an Arabidopsis mutant with impaired beta-oxidation, is defective in a peroxisomal beta-hydroxyisobutyryl-CoA hydrolase. AB - The Arabidopsis chy1 mutant is resistant to indole-3-butyric acid, a naturally occurring form of the plant hormone auxin. Because the mutant also has defects in peroxisomal beta-oxidation, this resistance presumably results from a reduced conversion of indole-3-butyric acid to indole-3-acetic acid. We have cloned CHY1, which appears to encode a peroxisomal protein 43% identical to a mammalian valine catabolic enzyme that hydrolyzes beta-hydroxyisobutyryl-CoA. We demonstrated that a human beta-hydroxyisobutyryl-CoA hydrolase functionally complements chy1 when redirected from the mitochondria to the peroxisomes. We expressed CHY1 as a glutathione S-transferase (GST) fusion protein and demonstrated that purified GST CHY1 hydrolyzes beta-hydroxyisobutyryl-CoA. Mutagenesis studies showed that a glutamate that is catalytically essential in homologous enoyl-CoA hydratases was also essential in CHY1. Mutating a residue that is differentially conserved between hydrolases and hydratases established that this position is relevant to the catalytic distinction between the enzyme classes. It is likely that CHY1 acts in peroxisomal valine catabolism and that accumulation of a toxic intermediate, methacrylyl-CoA, causes the altered beta-oxidation phenotypes of the chy1 mutant. Our results support the hypothesis that the energy-intensive sequence unique to valine catabolism, where an intermediate CoA ester is hydrolyzed and a new CoA ester is formed two steps later, avoids methacrylyl-CoA accumulation. PMID- 11404362 TI - The pH dependence of serpin-proteinase complex dissociation reveals a mechanism of complex stabilization involving inactive and active conformational states of the proteinase which are perturbable by calcium. AB - Serpin family protein proteinase inhibitors trap proteinases at the acyl intermediate stage of cleavage of the serpin as a proteinase substrate by undergoing a dramatic conformational change, which is thought to distort the proteinase active site and slow deacylation. To investigate the extent to which proteinase catalytic function is defective in the serpin-proteinase complex, we compared the pH dependence of dissociation of several serpin-proteinase acyl complexes with that of normal guanidinobenzoyl-proteinase acyl-intermediate complexes. Whereas the apparent rate constant for dissociation of guanidinobenzoyl-proteinase complexes (k(diss, app)) showed a pH dependence characteristic of His-57 catalysis of complex deacylation, the pH dependence of k(diss, app) for the serpin-proteinase complexes showed no evidence for His-57 involvement in complex deacylation and was instead characteristic of a hydroxide mediated deacylation similar to that observed for the hydrolysis of tosylarginine methyl ester. Hydroxylamine enhanced the rate of serpin-proteinase complex dissociation but with a rate constant for nucleophilic attack on the acyl bond several orders of magnitude slower than that of hydroxide, implying limited accessibility of the acyl bond in the complex. The addition of 10-100 mm Ca(2+) ions stimulated up to 80-fold the dissociation rate constant of several serpin trypsin complexes in a saturable manner at neutral pH and altered the pH dependence to a pattern characteristic of His-57-catalyzed complex deacylation. These results support a mechanism of kinetic stabilization of serpin-proteinase complexes wherein the complex is trapped as an acyl-intermediate by a serpin conformational change-induced inactivation of the proteinase catalytic function, but suggest that the inactive proteinase conformation in the complex is in equilibrium with an active proteinase conformation that can be stabilized by the preferential binding of an allosteric ligand such as Ca(2+). PMID- 11404363 TI - Affinity, kinetics, and thermodynamics of E-selectin binding to E-selectin ligand 1. AB - E-selectin is an endothelial adhesion molecule, which mediates the tethering and rolling of leukocytes on vascular endothelium. It recognizes the glycoprotein E selectin ligand-1 (ESL-1) as a major binding partner on mouse myeloid cells. Using surface plasmon resonance, we measured the kinetics and affinity of binding of monomeric E-selectin to ESL-1 isolated from mouse bone marrow cells. E selectin bound to ESL-1 with a fast dissociation rate constant of 4.6 s(-1) and a calculated association rate constant of 7.4 x 10(4) m(-1) s(-1). We determined a dissociation constant (K(d)) of 62 microm, which resembles the affinity of L selectin binding to glycosylation-dependent cell adhesion molecule-1. The affinity of the E-selectin-ESL-1 interaction did not change significantly when the temperature was varied from 5 degrees C to 37 degrees C, indicating that the enthalpic contribution to the binding is small at physiological temperatures, and that, in contrast to typical protein-carbohydrate interactions, binding is driven primarily by favorable entropic changes. Interestingly, surface plasmon resonance experiments with recombinant ESL-1 from alpha 1,3-fucosyltransferase IV expressing Chinese hamster ovary cells showed a very similar K(d) of 66 microm, suggesting that this fucosyltransferase is sufficient to produce fully functional recombinant ESL-1. Following the recent description of the affinity and kinetics of the selectin-ligand pairs L-selectin-glycosylation-dependent cell adhesion molecule-1 and P-selectin-P-selectin glycoprotein ligand-1, this is the first determination of the parameters of E-selectin binding to one of its naturally occurring ligands. PMID- 11404364 TI - The yeast Pma1 proton pump: a model for understanding the biogenesis of plasma membrane proteins. PMID- 11404365 TI - Ion pumps in polarized cells: sorting and regulation of the Na+, K+- and H+, K+ ATPases. AB - The physiologic function of an ion transport protein is determined, in part, by its subcellular localization and by the cellular mechanisms that modulate its activity. The Na(+),K(+)-ATPase and the H(+),K(+)-ATPases are closely related members of the P-type family of ion transporting ATPases. Despite their homology, these pumps are sorted to different domains in polarized epithelial cells, and their enzymatic activities are subject to distinct regulatory pathways. The molecular signals responsible for these properties have begun to be elucidated. It appears that a complex array of inter- and intramolecular interactions govern trafficking, distribution, and catalytic capacities of these proteins. PMID- 11404366 TI - Transport ATPase trafficking minireview series. PMID- 11404367 TI - Gelatinase B functions as regulator and effector in leukocyte biology. AB - Matrix metalloproteinases (MMPs) form a family of enzymes with major actions in the remodeling of extracellular matrix (ECM) components. Gelatinase B (MMP-9) is the most complex family member in terms of domain structure and regulation of its activity. Gelatinase B activity is under strict control at various levels: transcription of the gene by cytokines and cellular interactions; activation of the pro-enzyme by a cascade of enzymes comprising serine proteases and other MMPs; and regulation by specific tissue inhibitors of MMPs (TIMPs) or by unspecific inhibitors, such as alpha2-macroglobulin. Thus, remodeling ECM is the result of the local protease load, i.e., the net balance between enzymes and inhibitors. Glycosylation has a limited effect on the net activity of gelatinase B, and in contrast to the all-or-none effect of enzyme activation or inhibition, it results in a higher-level, fine-tuning effect on the ECM catalysis by proteases in mammalian species. Fast degranulation of considerable amounts of intracellularly stored gelatinase B from neutrophils, induced by various types of chemotactic factors, is another level of control of activity. Neutrophils are first-line defense leukocytes and do not produce gelatinase A or TIMP. Thus, neutrophils contrast sharply with mononuclear leukocytes, which produce gelatinase A constitutively, synthesize gelatinase B de novo after adequate triggering, and overproduce TIMP-1. Gelatinase B is also endowed with functions other than cleaving the ECM. It has been shown to generate autoimmune neo epitopes and to activate pro-IL-1beta into active IL-1beta. Gelatinase B ablation in the mouse leads to altered bone remodeling and subfertility, results in resistance to several induced inflammatory or autoimmune pathologies, and indicates that the enzyme plays a crucial role in development and angiogenesis. The major human neutrophil chemoattractant, IL-8, stimulates fast degranulation of gelatinase B from neutrophils. Gelatinase B is also found to function as a regulator of neutrophil biology and to truncate IL-8 at the amino terminus into a tenfold more potent chemokine, resulting in an important positive feedback loop for neutrophil activation and chemotaxis. The CXC chemokines GRO-alpha, CTAP-III, and PF-4 are degraded by gelatinase B, whereas the CC chemokines MCP-2 and RANTES are not cleaved. PMID- 11404368 TI - Novel chemokine functions in lymphocyte migration through vascular endothelium under shear flow. AB - The recruitment of circulating leukocytes at vascular sites in target tissue has been linked to activation of Gi-protein signaling in leukocytes by endothelial chemokines. The mechanisms by which apical and subendothelial chemokines regulate leukocyte adhesion to and migration across endothelial barriers have been elusive. We recently found that endothelial chemokines not only stimulate integrin-mediated arrest on vascular endothelial ligands but also trigger earlier very late antigen (VLA)-4 integrin-mediated capture (tethering) of lymphocytes to vascular cell adhesion molecule 1 (VCAM-1)-bearing surfaces by extremely rapid modulation of integrin clustering at adhesive contact zones. This rapid modulation of integrin avidity requires chemokine immobilization in juxtaposition with the VLA-4 ligand VCAM-1. We also observed that endothelial-bound chemokines promote massive lymphocyte transendothelial migration (TEM). It is interesting that chemokine-promoted lymphocyte TEM requires continuous exposure of lymphocytes but not of the endothelial barrier to fluid shear. It is noteworthy that lymphocyte stimulation by soluble chemokines did not promote lymphocyte TEM. Our results suggest new roles for apical endothelial chemokines both in triggering lymphocyte capture to the endothelial surface and in driving post arrest events that promote lymphocyte transmigration across endothelial barriers under shear flow. PMID- 11404369 TI - Role of activin A in negative regulation of normal and tumor B lymphocytes. AB - Activin A, a member of the transforming growth factor beta superfamily, has a wide spread expression pattern and pleiotropic functions. In this overview we summarize data that points to a role of activin A in negative regulation of B lineage lymphocytes. Experiments performed by us and by other groups revealed the capacity of activin A to cause apoptotic death of tumor myeloma cells, through mechanisms of cell cycle inhibition and antagonism with the survival signal of interleukin-6. In vitro studies on B lymphocyte generation from bone marrow stem cells and use of human nasal polyps as a model of inflamed tissue further demonstrate an inhibitory role of activin A in B cell spread and accumulation. These data are analyzed with respect to our model of tissue organization that we term the "restrictin model of cell growth regulation." This model assumes a morphogen-like role of activin A in the hematopoietic system. Thus, the relative concentration of biologically functional activin A, in different parts of the tissue, may determine the local B cell content and functional state of these cells within a specific microenvironment. PMID- 11404370 TI - Signaling networks regulating beta1 integrin-mediated adhesion of T lymphocytes to extracellular matrix. AB - T-cell recognition of foreign antigen and migration to specific anatomic sites in vivo involves transient adhesive contacts between beta1 integrins expressed on T cells and cell surface proteins or extracellular-matrix components. Engagement of the CD3-T-cell receptor (CD3-TCR) complex initiates a complex signaling cascade involving coordinated regulation and recruitment of tyrosine and lipid kinases to specific regions or microdomains in the plasma membrane. Although considerable attention has been focused on the signaling events by which the CD3-TCR complex regulates transcriptional events in the nucleus, CD3-TCR signaling also rapidly enhances integrin-mediated adhesion without increasing surface expression of integrins. Recent studies suggest that CD3-TCR signaling to beta1 integrins involves coordinated recruitment and activation of the Tec family tyrosine kinase Itk by src family tyrosine kinases and phosphatidylinositol 3-kinase. These signaling events that regulate integrin-mediated T-cell adhesion share both common and distinct features with the signaling pathways regulating interleukin-2 gene transcription. PMID- 11404371 TI - Agonistic and antagonistic activities of chemokines. AB - Since the discovery of interleukin-8, about 50 chemokines have been identified and characterized. Originally, they were considered as inducible mediators of inflammation, but in recent years, several chemokines were identified that are expressed constitutively and function in physiological traffic and homing of leukocyte-lymphocytes in particular. All chemokines act via seven-transmembrane domain, G protein-coupled receptors. Eighteen such receptors have been identified so far. Studies on structure-activity relationships indicate that chemokines have two main sites of interaction with their receptors, the flexible NH2-terminal region and the conformationally rigid loop that follows the second cysteine. Chemokines are thought to dock onto receptors by means of the loop region, and this contact is believed to facilitate the binding of the NH2-terminal region that results in receptor activation. These studies have also highlighted the importance of the NH2-terminal region for agonistic and antagonistic activity. Recently, we have shown that some naturally occurring chemokines can function as receptor antagonists. These observations suggest a new mechanism for the regulation of leukocyte recruitment during inflammatory and immune reactions, which are based on the combination of agonistic and antagonistic effects. PMID- 11404372 TI - Combinatorial signals by inflammatory cytokines and chemokines mediate leukocyte interactions with extracellular matrix. AB - On their extravasation from the vascular system into inflamed tissues, leukocytes must maneuver through a complex insoluble network of molecules termed the extracellular matrix (ECM). Leukocytes navigate toward their target sites by adhering to ECM glycoproteins and secreting degradative enzymes, while constantly orienting themselves in response to specific signals in their surroundings. Cytokines and chemokines are key biological mediators that provide such signals for cell navigation. Although the individual effects of various cytokines have been well characterized, it is becoming increasingly evident that the mixture of cytokines encountered in the ECM provides important combinatorial signals that influence cell behavior. Herein, we present an overview of previous and ongoing studies that have examined how leukocytes integrate signals from different combinations of cytokines that they encounter either simultaneously or sequentially within the ECM, to dynamically alter their navigational activities. For example, we describe our findings that tumor necrosis factor (TNF)-alpha acts as an adhesion-strengthening and stop signal for T cells migrating toward stromal cell-derived factor-1alpha, while transforming growth factor-beta down-regulates TNF-alpha-induced matrix metalloproteinase-9 secretion by monocytes. These findings indicate the importance of how one cytokine, such as TNF-alpha, can transmit diverse signals to different subsets of leukocytes, depending on its combination with other cytokines, its concentration, and its time and sequence of exposure. The combinatorial effects of multiple cytokines thus affect leukocytes in a step-by-step manner, whereby cells react to cytokine signals in their immediate vicinity by altering their adhesiveness, directional movement, and remodeling of the ECM. PMID- 11404373 TI - Shape and shift changes related to the function of leukocyte integrins LFA-1 and Mac-1. AB - Integrin activity on leukocytes is controlled tightly, ensuring that ligand binding occurs only when leukocytes are in contact with their targets. For an integrinlike LFA-1, this ligand-binding activity comes about as a result of increased integrin clustering. Affinity regulation of integrins also plays a role, but the conformational changes giving rise to increased affinity appear to be secondary to clustering. Conformationally altered LFA-1 can be created artificially by deletion of the I domain, which is the key domain involved in ligand binding for many but not all integrins. Although I domain-deleted LFA-1 (DeltaI-LFA-1) cannot bind ligand, it is able to signal constitutively into the cell. One measure of this signaling activity is the ability of DeltaI-LFA-1 to activate beta1 integrins on the same T lymphocyte. Leukocytes use LFA-1 to migrate across the endothelium. Active beta1 integrins may be required subsequently to bind the matrix proteins encountered by leukocytes as they continue their voyage into the tissue interior. PMID- 11404374 TI - Neutrophil recruitment, chemokine receptors, and resistance to mucosal infection. AB - Neutrophil migration to infected mucosal sites involves a series of complex interactions with molecules in the lamina propria and at the epithelial barrier. Much attention has focussed on the vascular compartment and endothelial cells, but less is known about the molecular determinants of neutrophil behavior in the periphery. We have studied urinary tract infections (UTIs) to determine the events that initiate neutrophil recruitment and interactions of the recruited neutrophils with the mucosal barrier. Bacteria activate a chemokine response in uroepithelial cells, and the chemokine repertoire depends on the bacterial virulence factors and on the specific signaling pathways that they activate. In addition, epithelial chemokine receptor expression is enhanced. Interleukin (IL) 8 and CXCR1 direct neutrophil migration across the epithelial barrier into the lumen. Indeed, mIL-8Rh knockout mice showed impaired transepithelial neutrophil migration, with tissue accumulation of neutrophils, and these mice developed renal scarring. They had a defective antibacterial defense and developed acute pyelonephritis with bacteremia. Low CXCR1 expression was also detected in children with acute pyelonephritis. These results demonstrate that chemokines and chemokine receptors are essential to orchestrate a functional antimicrobial defense of the urinary tract mucosa. Mutational inactivation of the IL-8R caused both acute disease and chronic tissue damage. PMID- 11404375 TI - Control of chromatin accessibility for V(D)J recombination by interleukin-7. AB - IL-7 is a key factor for lymphoid development, and it contributes to V(D)J recombination at multiple loci in immune-receptor genes. IL-7 signal transduction, involving gamma(c) and Jak3, is required for successful recombination at the TCR-gamma locus. IL-7 signaling controls the initiation phase of V(D)J recombination by controlling access of the V(D)J recombinase to the locus. In the absence of IL-7, the TCR-gamma locus is methylated and packaged in a repressed form of chromatin consisting of hypoacetylated histones. IL-7 signaling likely increases the acetylation state of the nucleosomal core histones resulting in an "open" form of chromatin. This opening leads to a higher accessibility for the transcription machinery and increased accessibility of the Rag heterodimer that performs the cleavage of DNA. PMID- 11404376 TI - Functional classification of interferon-stimulated genes identified using microarrays. AB - Interferons (IFNs) are a family of multifunctional cytokines that activate transcription of subsets of genes. The gene products induced by IFNs are responsible for IFN antiviral, antiproliferative, and immunomodulatory properties. To obtain a more comprehensive list and a better understanding of the genes regulated by IFNs, we compiled data from many experiments, using two different microarray formats. The combined data sets identified >300 IFN stimulated genes (ISGs). To provide new insight into IFN-induced cellular phenotypes, we assigned these ISGs to functional categories. The data are accessible on the World Wide Web at http://www.lerner.ccf.org/labs/williams/, including functional categories and individual genes listed in a searchable database. The entries are linked to GenBank and Unigene sequence information and other resources. The goal is to eventually compile a comprehensive list of all ISGs. Recognition of the functions of the ISGs and their specific roles in the biological effects of IFNs is leading to a greater appreciation of the many facets of these intriguing and essential cytokines. This review focuses on the functions of the ISGs identified by analyzing the microarray data and focuses particularly on new insights into the protein kinase RNA-regulated (PRKR) protein, which have been made possible with the availability of PRKR-null mice. PMID- 11404377 TI - Superantigen antagonist blocks Th1 cytokine gene induction and lethal shock. AB - Bacterial superantigens trigger an excessive, Th1-cytokine response leading to toxic shock. We designed a peptide antagonist that inhibits SEB-induced expression of human genes for IL-2, IFN-gamma, and TNF-beta, cytokines that mediate shock. The peptide antagonist shows homology to a beta-strand-hinge-alpha helix domain that is conserved structurally in superantigens produced by Staphylococcus aureus and Streptococcus pyogenes yet remote from known binding sites for the major histocompatibility class II molecule and T-cell receptor. For Th1-cell activation, superantigens depend on this domain. The peptide protected mice against lethal challenge with SEB or SEA. Moreover, it rescued mice undergoing toxic shock. Surviving mice rapidly developed broad-spectrum, protective immunity, which rendered them resistant to further lethal challenges with different staphylococcal and streptococcal superantigens. Thus, the lethal effect of superantigens, mediated by Th1 cytokines, can be blocked with a peptide antagonist that inhibits their action at the top of the toxicity cascade, before activation of T cells takes place. PMID- 11404378 TI - TSG-14 transgenic mice have improved survival to endotoxemia and to CLP-induced sepsis. AB - Tumor necrosis factor-stimulated gene 14 (TSG-14)/PTX3 was identified originally as a TNF-alpha and IL-1beta-stimulated gene from normal, human foreskin fibroblasts and vascular endothelial cells, respectively. TSG-14 gene encodes a 42-kDa-secreted glycoprotein with a carboxy-terminal half that shares homology with the entire sequence of C-reactive protein (CRP) and serum amyloid P component (SAP), acute-phase proteins of the pentraxin family. Some experimental evidence suggests that TSG-14 plays a role in inflammation, yet its function and mechanism of action remain unclear. We have generated transgenic mice that overexpress the murine TSG-14 gene under the control of its own promoter. From eight transgenic founders, two lineages were derived and better characterized: Tg2 and Tg4, carrying two and four copies of the transgene, respectively. TSG-14 transgenic mice were found to be more resistant to the endotoxic shock induced by LPS and to the polymicrobial sepsis caused by cecal ligation and puncture (CLP). Moreover, macrophages derived from the transgenic mice produced higher amounts of nitric oxide in response to IFN-gamma, TNF-alpha, and LPS as compared with macrophages from wild-type animals, and the augmented response appears to be the consequence of a higher responsiveness of transgenic macrophages to IFN-gamma. The data shown here are the first in vivo evidence of the involvement of TSG-14 in the inflammatory process and suggest a role for TSG-14 in the defense against bacterial infections. PMID- 11404379 TI - Transient expansion of peptide-specific lymphocytes producing IFN-gamma after vaccination with dendritic cells pulsed with MAGE peptides in patients with mage A1/A3-positive tumors. AB - Assessment of T-cell activation is pivotal for evaluation of cancer immunotherapy. We initiated a clinical trial in patients with MAGE-A1 and/or -A3 tumors using autologous DC pulsed with MAGE peptides aimed at analyzing T-cell derived, IFN-gamma secretion by cytokine flow cytometry and ELISPOT. We also tested whether further KLH addition could influence this response favorably. Monocyte-derived DC were generated from leukapheresis products. They were pulsed with the relevant MAGE peptide(s) alone in group A (n=10 pts) and additionally with KLH in group B (n=16 pts). A specific but transient increase in the number of peripheral blood T lymphocytes secreting IFN-gamma in response to the vaccine peptide(s) was observed in 6/8 patients of group A and in 6/16 patients of group B. We conclude that anti-tumor vaccination using DC pulsed with MAGE peptides induces a potent but transient anti-MAGE, IFN-gamma secretion that is not influenced by the additional delivery of a nonspecific, T-cell help. PMID- 11404380 TI - Tissue distribution of CD6 and CD6 ligand in cattle: expression of the CD6 ligand (CD166) in the autonomic nervous system of cattle and the human. AB - We studied the tissue distribution of CD6(+) lymphocytes and cells expressing the CD6 ligand (also known as activated leukocyte cell adhesion molecule CD166) in calves by immunohistochemistry using an anti-bovine CD6 monoclonal antibody (mAb), a human CD6 (huCD6)-immunoglobulin G1 fusion protein (huCD6-Ig), and an anti-human CD166 (anti-huCD166) mAb. The huCD6-Ig and anti-huCD166 mAb bound to the sympathetic and parasympathetic nerve fibers but not to myelinated nerve fibers in the spinal nerve. Studies with human tissue using the anti-huCD166 mAb yielded identical patterns of labeling. Dense accumulations of CD6(+) lymphocytes were present in areas of the thymuses and spleens of calves, in areas innervated by huCD6-Ig(+) nerves. The cDNAs encoding the bovine CD166 and CD6 were isolated from the sympathetic ganglion and spleen, respectively. Predicted amino acid residues that are important for human and mouse CD6-CD166 binding were also conserved in bovine CD6 and CD166. Bovine CD166 transcripts were detected by reverse transcriptase-PCR in all the tissues that bound huCD6-Ig. These results show that the bovine orthologue of CD166 was constitutively expressed in the autonomic nervous systems of cattle and suggest that CD6(+) lymphocytes adhere to CD166(+) autonomic nerve terminals via CD6. PMID- 11404381 TI - CD8alpha(-) and CD8alpha(+) subclasses of dendritic cells undergo phenotypic and functional maturation in vitro and in vivo. AB - Dendritic cells (DCs) are essential for the priming of immune responses. This antigen-presenting function of DCs develops in sequence in a process called maturation, during which they become potent sensitizers of naive T cells but reduce their ability to capture and process antigens. Some heterogeneity exists in mouse-DC populations, and two distinct subsets of DCs expressing high levels of CD11c can be identified on the basis of CD8alpha expression. We have studied the phenotype and maturation state of mouse splenic CD8alpha(-) and CD8alpha(+) DCs. Both subsets were found to reside in the spleen as immature cells and to undergo a phenotypic maturation upon culture in vitro in GM-CSF-containing medium or in vivo in response to lipopolysaccharide. In vitro and in vivo analyses showed that this maturation process is an absolute requisite for DCs to acquire their T-cell priming capacity, transforming CD8alpha(-) and CD8alpha(+) DCs into potent and equally efficient activators of naive CD4(+) and CD8(+) T cells. Furthermore, these results highlight the importance that environmental factors may have on the ability of DC subsets to influence Th responses qualitatively; i.e., the ability to drive Th1 versus Th2 differentiation may not be fixed immutably for each DC subset. PMID- 11404382 TI - Statins suppress THP-1 cell migration and secretion of matrix metalloproteinase 9 by inhibiting geranylgeranylation. AB - Macrophages secrete matrix metalloproteinase 9 (MMP-9), an enzyme that weakens the fibrous cap of atherosclerotic plaques, predisposing them to plaque rupture and subsequent ischemic events. Recent work indicates that statins strongly reduce the possibility of heart attack. Furthermore, these compounds appear to exert beneficial effects not only by lowering plasma low-density-lipoprotein cholesterol but also by directly affecting the artery wall. To evaluate whether statins influence the proinflammatory responses of monocytic cells, we studied their effects on the chemotactic migration and MMP-9 secretion of human monocytic cell line THP-1. Simvastatin dose dependently inhibited THP-1 cell migration mediated by monocyte chemoattractant protein 1, with a 50% inhibitory concentration of about 50 nM. It also inhibited bacterial lipopolysaccharide stimulated secretion of MMP-9. The effects of simvastatin were completely reversed by mevalonate and its derivatives, farnesylpyrophosphate and geranylgeranyl pyrophosphate, but not by ubiquinone. Additional studies revealed similar but more profound inhibitory effects with L-839,867, a specific inhibitor of geranylgeranyl transferase. However, alpha-hydroxyfarnesyl phosphonic acid, an inhibitor of farnesyl transferase, had no effect. C3 exoenzyme, a specific inhibitor of the prenylated small signaling Rho proteins, mimicked the inhibitory effects of simvastatin and L-839,867. These data supported the role of geranylgeranylation in the migration and MMP-9 secretion of monocytes. PMID- 11404383 TI - Involvement of cytosolic prolyl endopeptidase in degradation of p40-phox splice variant protein in myeloid cells. AB - Our previous studies indicated that an alternatively spliced variant mRNA of p40 phox, a cytosolic component of NADPH oxidase, is expressed but its protein is hardly detected in myeloid cells such as promyelocytic HL-60 cells and neutrophils. Here, we have examined the stability of p40-phox variant protein in undifferentiated HL-60 cells. When in vitro-translated proteins were incubated with subcellular fractions of HL-60 cells, p40-phox variant protein but not native p40-phox was degraded by the cytosol and granule fractions. The degradation of variant protein by the granule fraction was observed using sonicated but not intact granules, suggesting that the variant protein is unlikely to be degraded by the granules in intact cells. To identify the enzyme(s) involved, we examined the effects of various enzyme inhibitors on the degradation of variant protein by the cytosol fraction. Degradation was completely inhibited by proline-specific serine protease (prolyl endopeptidase) inhibitors but not by proteasome, calpain, and metalloprotease inhibitors. Furthermore, the variant protein was degraded by a purified prolyl endopeptidase, and the degradation was protected by treating HL-60 cells with a cell-permeable inhibitor (S17092-1) for prolyl endopeptidase. These observations suggest that a cytosolic prolyl endopeptidase is involved in the degradation of p40-phox variant protein in myeloid cells. PMID- 11404384 TI - Differences in splenic B-lymphocyte ganglioside expression and accessibility in normal and endotoxin-hyporesponsive mice. AB - Endotoxin-responsive (C3H/HeN) and -hyporesponsive (C3H/HeJ) murine B lymphocytes purified by adherence to anti-immunoglobulin ("antibody panning") possess identical gangliosides but different ganglioside surface accessibilities. We investigated the distribution and surface accessibility of gangliosides of B lymphocytes purified by adherence to plastic ("plastic panning") or by subtraction of non-B-lymphocyte components. As with antibody panning, there were no entirely new or absent gangliosides in plastic-panned or subtraction-purified B lymphocytes of each strain. However, striking changes in relative expression of five gangliosides were detected with each purification protocol. Moreover, five gangliosides of antibody-panned and plastic-panned B lymphocytes but only two gangliosides of subtraction-purified B lymphocytes were inaccessible to surface labeling. Unlike the situation for antibody-panned B lymphocytes, no interstrain (HeN vs. HeJ) surface accessibility differences existed in gangliosides of plastic-panned or subtraction-purified cells. Exposure of subtraction-purified B lymphocytes to anti-immunoglobulin failed to elicit changes in ganglioside expression. Murine B lymphocytes have distinct protocol-dependent differences in glycolipid phenotype which likely denote individual subpopulations. PMID- 11404385 TI - A chimeric MIP-1alpha/RANTES protein demonstrates the use of different regions of the RANTES protein to bind and activate its receptors. AB - Human RANTES (CCL5) and MIP-1alpha (CCL3) bind and activate several CC chemokine receptors. RANTES is a high-affinity ligand for CCR1 and CCR5, and it binds CCR3 with moderate affinity and CCR4 with low affinity. MIP-1alpha has similar binding characteristics to RANTES except that it does not bind to CCR3. Here we have generated a chimera of human MIP-1alpha and RANTES, called MIP/RANTES, consisting of the eight amino terminal residues of MIP-1alpha preceding the CC motif, and the remainder of the sequence is RANTES. The chimera is able to induce chemotaxis of human monocytes. MIP/RANTES has >100-fold reduction in binding to CCR1 and does not bind to CCR3 but retains full, functional binding to CCR5. It has equivalent affinity for CCR5 to MIP-1alpha and RANTES, binding with an IC(50) of 1.12 nM, and is able to mobilize calcium and induce endocytosis of CCR5 in PBMC in a manner equi-potent to RANTES. It also retains the ability to inhibit R5 using HIV-1 strains. Therefore, we conclude that the amino terminus of RANTES is not involved in CCR5 binding, but it is essential for CCR1 and CCR3. PMID- 11404386 TI - Proinflammatory properties of the human S100 protein S100A12. AB - S100 proteins represent a new class of chemoattractants. Here we extend earlier evidence for the proinflammatory properties of human S100A12. A12 induced migration of monocytoid cells, with optimal activity at 10(-10) M and potency of >10(-9) M C5a. Neutrophils were poorly responsive, and lymphocyte migration was not affected. Actin polymerization in monocytoid cells was accompanied by a sustained [Ca(2+)]i flux of a magnitude comparable with C5a. A12 elicited a transient infiltration of neutrophils (4-8 h) and more delayed recruitment of monocytes (8-24 h) in vivo. A12 (approximately 70 nM) was present in synovial fluid (SF) from rheumatoid arthritis patients, and synovium contained A12 positive neutrophils in the sublining and interstitial region, often surrounding the perivasculature but rarely in the synovial lining layer, although some macrophages were positive. The A12 gene was transiently up-regulated in monocytes by tumor necrosis factor alpha (6 h); induction by lipopolysaccharide (LPS) was sustained (12-48 h). A12 may contribute to leukocyte migration in chronic inflammatory responses. PMID- 11404387 TI - Expression, tissue distribution, and cellular localization of the antiapoptotic TIP-B1 protein. AB - TIP-B1 is a novel 27-kDa protein isolated from the cytosol of tumor necrosis factor (TNF)-stimulated cells. Cells preincubated with TIP-B1 are protected from TNF-induced apoptosis. This study showed that, as with normal fibroblasts and U937 histiocytic lymphoma, human MCF7 mammary adenocarcinoma cells were protected from TNF in a concentration-dependent manner by pretreatment with either TNF or purified TIP-B1. Immunoblot and immunohistochemical analyses indicated expression of both TIP-B1 mRNA and protein in MCF7 cells and heart, kidney, brain, liver, ovary, uterus, thymus, spleen, lymph node, and mammary gland cells throughout their development. Expression of TIP-B1 was heterogeneous, with staining of specific cell types within tissues. Based on the ability of TIP-B1 to protect both normal and tumor cells from TNF-induced apoptosis and its broad tissue distribution, with expression only in select cells within those tissues, a role for TIP-B1 in the regulation of TNF-induced effects is strongly indicated. PMID- 11404388 TI - Mechanisms of neutrophil apoptosis in uremia and relevance of the Fas (APO-1, CD95)/Fas ligand system. AB - The regulation of neutrophil apoptosis in chronic renal failure (CRF) has not been clearly defined. The Fas/FasL system is an important apoptotic regulatory pathway in a wide variety of cells. Fas is a widely expressed cell surface protein that transduces an apoptotic signal after interaction with its natural ligand FasL. In contrast to the extensive tissue distribution of Fas, constitutive expression of FasL is relatively limited. We examined Fas and FasL expression by neutrophils in healthy subjects, patients with CRF, and patients on hemodialysis (HD) and peritoneal dialysis (PD). Fas expression was significantly higher among patients with CRF compared with control subjects, HD patients, and PD patients. FasL expression was significantly higher among patients with CRF compared with control subjects. At 24 h, neutrophil apoptosis was higher among patients with CRF compared with control subjects. Furthermore, high-neutrophil Fas expression was paralleled by a higher sensitivity to Fas-mediated apoptosis. There was a strong correlation between Fas-stimulated apoptosis and creatinine clearance as well as Fas expression. Finally, we found that uremic serum increased the expression of neutrophil-associated Fas and FasL proteins, when compared with normal serum. Further studies are under way to examine the regulation of this pathway in the uremic environment. PMID- 11404389 TI - Priming effects of substance P on calcium changes evoked by interleukin-8 in human neutrophils. AB - The neurokinin (NK) substance P (SP), which is a mediator of neurogenic inflammation, has been reported to prime human polymorphonuclear neutrophils (PMNs). The priming effects of SP on PMNs activated by recombinant interleukin-8 (rIL-8) were investigated. SP enhanced, in a dose- and time-dependent way, the rise in cytosolic free-calcium concentration, [Ca(2+)]i, evoked by the chemokine. The priming effects of SP were abolished by exposing PMNs to a calcium-free medium supplemented with EGTA. The C-terminal peptides SP(4-11) and SP(6-11) but not the N-terminal peptide SP(1-7) shared the priming effects of SP. The selective NK-1 receptor agonist [Sar-9, MetO2-11]SP mimicked the effects of SP, which were not reproduced by the selective NK-2 receptor agonist [betaAla-8] NKA(4-10) or the selective NK-3 agonist senktide. Two selective NK-1 antagonists, CP96,345 and L703,606, dose dependently inhibited SP priming effects. These results demonstrated that SP primes PMNs exposed to rIL-8 and suggested that SP priming effects are receptor mediated. PMID- 11404390 TI - Nerve growth factor regulates TNF-alpha production in mouse macrophages via MAP kinase activation. AB - In this study, we examined the expression of nerve growth factor (NGF) and its receptors in mouse macrophages and the mechanisms involved in the effect of NGF on tumor necrosis factor (TNF)-alpha production. Macrophages expressed NGF and the NGF receptors TrkA and p75. Treatment of J744 cells or peritoneal macrophages with NGF induced a large increase in the production of TNF-alpha. In addition, NGF induced the secretion of nitric oxide in interferon-gamma-treated J774 cells or lipopolysaccharide-treated peritoneal macrophages. The induction of TNF-alpha production by NGF was blocked by K252a, an inhibitor of the TrkA receptor. NGF induced phosphorylation and activation of extracellular signal-regulated kinase, Erk1/Erk2 and c-Jun amino-terminal kinase, whereas it did not induce phosphorylation of p38 mitogen-activated protein kinase. Inhibition of the MAP kinase-Erk kinase pathway with PD 098059 decreased the secretion of TNF-alpha by NGF. Our results suggest that NGF has an important role in the activation of macrophages during inflammatory responses via activation of mitogen-activated protein kinases. PMID- 11404391 TI - Identification and characterization of human eosinophil cationic protein by an epitope-specific antibody. AB - The eosinophil cationic protein (ECP) is a basic secretion protein involved in the immune response system. ECP levels in biological fluids are an indicator of eosinophil-specific activation and degranulation and are currently used for the clinical monitoring and diagnosis of inflammatory disorders. A polyclonal epitope specific antibody has been obtained by immunizing rabbits with a conjugated synthetic peptide. A sequence corresponding to a large exposed loop in the human ECP three-dimensional structure (D115-Y122) was selected as a putative antigenic epitope. The antibody was purified on an affinity column using recombinant ECP (rECP) as antigen. The antibody (D112-P123 Ab) specifically recognizes rECP and its native glycosylated and nonglycosylated forms in plasma, granulocytes, and sputum. The antibody detects as little as 1 ng of rECP, can be used both in reducing and nonreducing conditions, and does not cross-react with the highly homologous eosinophil-derived neurotoxin or other proteins of the pancreatic ribonuclease superfamily. PMID- 11404392 TI - Different Toll-like receptor agonists induce distinct macrophage responses. AB - We previously reported that gram-negative bacterial lipopolysaccharide (LPS) activates cells via Toll-like receptor (TLR) 4, whereas the mycobacterial cell wall glycolipid lipoarabinomannan (LAM) activates cells via TLR2. We also identified a secreted TLR2 agonist activity in short-term culture filtrates of Mycobacterium tuberculosis bacilli, termed soluble tuberculosis factor (STF). Here we show that STF contains mannosylated phosphatidylinositol (PIM) and that purified PIM possesses TLR2 agonist activity. Stimulation of RAW 264.7 macrophages by LPS, LAM, STF, and PIM rapidly activated nuclear factor (NF) kappaB, activator protein-1 (AP-1), and mitogen-activated protein (MAP) kinases. These TLR agonists induced similar levels of NF-kappaB and AP-1 DNA-binding activity, as well as trans-activation function. Unexpectedly, these TLR agonists induced tumor necrosis factor alpha secretion, whereas only LPS was capable of inducing interleukin-1beta and nitric oxide secretion. Thus, different TLR proteins are still capable of activating distinct cellular responses, in spite of their shared capacities to activate NF-kappaB, AP-1, and MAP kinases. PMID- 11404393 TI - Cloning and expression analysis of a novel G-protein-coupled receptor selectively expressed on granulocytes. AB - The migration of neutrophils into sites of acute and chronic inflammation is mediated by chemokines. We used degenerate-primer reverse transcriptase polymerase chain reaction (RT-PCR) to analyze chemokine receptor expression in neutrophils and identify novel receptors. RNA was isolated from human peripheral blood neutrophils and from neutrophils that had been stimulated for 5 h with granulocyte-macrophage colony-stimulating factor or by coculturing with primary human bronchial epithelial cells. Amplification products were cloned, and clone redundancy was determined. Seven known G-protein-coupled receptors were identified among 38 clones-CCR1, CCR4, CXCR1, CXCR2, CXCR4, HM63, and FPR1-as well as a novel gene, EX33. The full-length EX33 clone was obtained, and an in silico approach was used to identify the putative murine homologue. The EX33 gene encodes a 396-amino-acid protein with limited sequence identity to known receptors. Expression studies of several known chemokine receptors and EX33 revealed that resting neutrophils expressed higher levels of CXCRs and EX33 compared with activated neutrophils. Northern blot experiments revealed that EX33 is expressed mainly in bone marrow, lung, and peripheral blood leukocytes. Using RT-PCR analysis, we showed more abundant expression of EX33 in neutrophils and eosinophils, in comparison with that in T- or B-lymphocytes, indicating cell specific expression among leukocytes. PMID- 11404394 TI - Suppression of T-cell responsiveness by inducible cAMP early repressor (ICER). AB - Depending on the nature of the costimulation of T lymphocytes, expression of regulatory cytokines and chemokines is either susceptible or resistant to cyclic AMP (cAMP)-mediated inhibition. Our data show that cAMP-mediated inhibition of endogenously expressed cytokines, which is characteristic for T helper (Th) 1- and Th 2-like phenotypes, correlates with the induction of a potent transcriptional repressor, inducible cAMP early repressor (ICER), in both subsets of T cells activated under conditions of suboptimal interleukin-2 (IL-2) expression. Importantly, Th-specific expression of certain chemokines is also susceptible to cAMP-mediated transcriptional attenuation. To determine whether ICER per se, rather than forskolin-mediated elevation of intracellular cAMP, is responsible for the observed inhibitory effect, we generated transgenic mice expressing ICER under the control of a lymphocyte-specific lck promoter. On stimulation, transgenic thymocytes overexpressing ICER exhibited reduced levels of IL-2 and interferon (IFN)-gamma and failed to express the macrophage inflammatory protein (MIP)-1alpha and MIP-1beta genes. Splenic T cells from ICER transgenic mice showed a defect in proliferation and lacked a mixed lymphocyte reaction response, implying that ICER-mediated inhibition of cytokine and chemokine expression might play an important role in T-cell inactivation. PMID- 11404395 TI - Phosphatidylcholine-specific phospholipase C regulates activation of RAW264.7 macrophage-like cells by lipopeptide JBT3002. AB - Phospholipase activities are thought to be involved in the activation of macrophages by lipopolysaccharide (LPS). Because our previous studies showed that the synthetic lipopeptide JBT3002 might activate macrophages via signaling pathways similar to those used by LPS, we investigated whether phospholipase activities are required for activation of macrophages by JBT3002. Treatment of RAW264.7 murine macrophage-like cells with JBT3002 stimulated expression of both inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF alpha) in a dose-dependent manner. The JBT3002-induced production of nitric oxide and TNF-alpha was significantly inhibited by tricyclodecan-9-yl xanthogenate (D609), a selective inhibitor of phosphatidylcholine (PC)-specific phospholipase C (PC-PLC). JBT3002-induced expression of steady-state mRNA for both iNOS and TNF alpha was inhibited by D609. Cells treated with JBT3002 had greater production of diacylglycerol (DAG) in 2 min, which lasted for at least 30 min and could be blocked by D609. Activation of RAW264.7 cells was not affected by butanol, a PC specific phospholipase D inhibitor, and treatment with JBT3002 did not affect phosphatidic acid formation. RAW264.7 cells treated with DAG analogue 1-oleoyl-2 acetyl-sn-glycerol, in the presence of interferon-gamma, produced TNF-alpha. These results suggested that activation of RAW264.7 cells by JBT3002 requires PC PLC activity. PMID- 11404397 TI - Beta-amyloid activates the mitogen-activated protein kinase cascade via hippocampal alpha7 nicotinic acetylcholine receptors: In vitro and in vivo mechanisms related to Alzheimer's disease. AB - Alzheimer's Disease (AD) is the most common of the senile dementias, the prevalence of which is increasing rapidly, with a projected 14 million affected worldwide by 2025. The signal transduction mechanisms that underlie the learning and memory derangements in AD are poorly understood. beta-Amyloid (Abeta) peptides are elevated in brain tissue of AD patients and are the principal component of amyloid plaques, a major criterion for postmortem diagnosis of the disease. Using acute and organotypic hippocampal slice preparations, we demonstrate that Abeta peptide 1-42 (Abeta42) couples to the mitogen-activated protein kinase (MAPK) cascade via alpha7 nicotinic acetylcholine receptors (nAChRs). In vivo elevation of Abeta, such as that exhibited in an animal model for AD, leads to the upregulation of alpha7 nAChR protein. alpha7 nAChR upregulation occurs concomitantly with the downregulation of the 42 kDa isoform of extracellular signal-regulated kinase (ERK2) MAPK in hippocampi of aged animals. The phosphorylation state of a transcriptional mediator of long-term potentiation and a downstream target of the ERK MAPK cascade, the cAMP-regulatory element binding (CREB) protein, were affected also. These findings support the model that derangement of hippocampus signal transduction cascades in AD arises as a consequence of increased Abeta burden and chronic activation of the ERK MAPK cascade in an alpha7 nAChR-dependent manner that eventually leads to the downregulation of ERK2 MAPK and decreased phosphorylation of CREB protein. PMID- 11404398 TI - Extracellular nucleotides differentially regulate interleukin-1beta signaling in primary human astrocytes: implications for inflammatory gene expression. AB - The cytokine interleukin-1beta (IL-1beta) is a potent activator of human astrocytes, inducing or modulating expression of multiple proinflammatory genes via activation of the transcription factors nuclear factor-kappaB (NF-kappaB) and activator protein-1 (AP-1). In this study, we examined whether IL-1beta signaling is regulated in these cells by extracellular nucleotides that are released at high concentrations under inflammatory conditions and act as ligands for members of the P2 receptor family. Using reporter constructs and electromobility shift assays, we found that cotreatment of astrocyte cultures with ATP (1-100 microm) significantly potentiated IL-1beta-mediated activation of NF-kappaB and AP-1 and that ATP alone activated AP-1. These effects were blocked by the P2 receptor antagonists XAMR 0721, periodate-oxidized ATP, and suramin. A role for ATP in modulating IL-1beta-mediated inflammatory gene expression was supported further by the observation that ATP potentiated the IL-1beta-induced expression of IL-8 mRNA and protein but strongly downregulated IP-10 expression. Reverse transcription-PCR and cloning demonstrated expression of the ATP-responsive P2 receptor subtypes P2Y(1), P2Y(2), and P2X(7), as well as the ATP-insensitive receptor P2Y(4). ADP, a selective agonist for P2Y(1), produced results similar to or greater than those obtained using ATP, whereas 2'-3'-O-(4-benzoyl-benzoyl) ATP, a selective agonist for P2X(7), was less effective than ATP. In contrast, UTP, a selective agonist for P2Y(2) and P2Y(4), was ineffective. These studies indicate that different P2 receptor subtypes play distinct roles in the modulation of IL-1beta-mediated signal transduction in human astrocytes, and that signaling via P2 receptors may fine-tune the transcription of genes involved in inflammatory responses in the human CNS. PMID- 11404399 TI - A novel extracellular calcium sensing mechanism in voltage-gated potassium ion channels. AB - Potassium (K(+)) channels influence neurotransmitter release, burst firing rate activity, pacing, and critical dampening of neuronal circuits. Internal and external factors that further modify K(+) channel function permit fine-tuning of neuronal circuits. Human ether-a-go-go-related gene (HERG) K(+) channels are unusually sensitive to external calcium concentration ([Ca(2+)](o)). Small changes in [Ca(2+)](o) shift the voltage dependence of channel activation to more positive membrane potentials, an effect that cannot be explained by nonspecific surface charge screening or channel pore block. The HERG-calcium concentration response relationship spans the physiological range for [Ca(2+)](o). The modulatory actions of calcium are attributable to differences in the Ca(2+) affinity between rested and activated channels. Adjacent extracellular, negatively charged amino acids (E518 and E519) near the S4 voltage sensor influence both channel gating and Ca(2+) dependence. Neutralization of these charges had distinct effects on channel gating and calcium sensitivity. A change in the degree of energetic coupling between these amino acids on transition from closed to activated channel states reveals movement in this region during channel gating and defines a molecular mechanism for protein state-dependent ligand interactions. The results suggest a novel extracellular [Ca(2+)](o) sensing mechanism coupled to allosteric changes in channel gating and a mechanism for fine-tuning cell repolarization. PMID- 11404400 TI - Kinetic modulation of Kv4-mediated A-current by arachidonic acid is dependent on potassium channel interacting proteins. AB - The Kv4 subfamily of voltage-gated potassium channels is responsible for the transient A-type potassium current that operates at subthreshold membrane potentials to control membrane excitability. Arachidonic acid was shown recently to modulate both the peak amplitude and kinetics of the hippocampal A-current. However, in Xenopus oocytes, arachidonic acid only inhibited the peak amplitude of Kv4 current without modifying its kinetics. These results suggest the existence of Kv4 auxiliary subunit(s) in native cells. We report here a K-channel interacting protein (KChIP)-dependent kinetic modulation of Kv4.2 current in Chinese hamster ovary cells and Kv4.2 and Kv4.3 currents in Xenopus oocytes by arachidonic acid at physiological concentrations. This concentration-dependent effect of arachidonic acid resembled that observed in cerebellar granule neurons and was fully reversible. Other fatty acids, including a nonhydrolyzable inhibitor of both lipooxygenase and cyclooxygenase, 5,8,11,14-eicosatetraynoic acid (ETYA), also mimicked arachidonic acid in modulating Kv4.3 and Kv4.3/KChIP1 currents. Compared with another transient potassium current formed by Kv1.1/Kvbeta1, Kv4.3/KChIP1 current was much more sensitive to arachidonic acid. Association between KChIP1 and Kv4.2 or Kv4.3 was not altered in the presence of 10 microm ETYA as measured by immunoprecipitation and association-dependent growth in yeast. Our data suggest that the KChIP proteins represent a molecular entity for the observed difference between arachidonic acid effects on A-current kinetics in heterologous cells and in native cells and are consistent with the notion that KChIP proteins modulate the subthreshold A-current in neurons. PMID- 11404401 TI - Channel-lining residues of the AMPA receptor M2 segment: structural environment of the Q/R site and identification of the selectivity filter. AB - In AMPA receptor channels, a single amino acid residue (Q/R site) of the M2 segment controls permeation of calcium ions, single-channel conductance, blockade by intracellular polyamines, and permeation of anions. The structural environment of the Q/R site and its positioning with regard to a narrow constriction were probed with the accessibility of substituted cysteines to positively and negatively charged methanethiosulfonate reagents, applied from the extracellular and cytoplasmic sides of the channel. The accessibility patterns confirm that the M2 segment forms a pore loop with the Q/R site positioned at the tip of the loop (position 0) facing the extracellular vestibule. Cytoplasmically accessible residues on the N- and C-terminal sides of position 0 form the ascending alpha helical (-8 to -1) and descending random coil (+1 to +6) components of the loop, respectively. Substitution of a glycine residue at position +2 with alanine strongly decreased the permeability of organic cations, indicating that position +2 contributes to the narrow constriction. The anionic 2-sulfonatoethyl methanethiosufonate reacted with a cysteine at position 0 only from the external side and with cysteines at positions +1 to +4 only from the cytoplasmic side. These results suggest that charge selectivity occurs external to the constriction (+2) and possibly involves interactions of ions with the negative electrostatic potential created by the dipole of the alpha-helix formed by the ascending limb of the loop. PMID- 11404402 TI - Extracellular calcium modulates persistent sodium current-dependent burst-firing in hippocampal pyramidal neurons. AB - The generation of high-frequency spike bursts ("complex spikes"), either spontaneously or in response to depolarizing stimuli applied to the soma, is a notable feature in intracellular recordings from hippocampal CA1 pyramidal cells (PCs) in vivo. There is compelling evidence that the bursts are intrinsically generated by summation of large spike afterdepolarizations (ADPs). Using intracellular recordings in adult rat hippocampal slices, we show that intrinsic burst-firing in CA1 PCs is strongly dependent on the extracellular concentration of Ca(2+) ([Ca(2+)](o)). Thus, lowering [Ca(2+)](o) (by equimolar substitution with Mn(2+) or Mg(2+)) induced intrinsic bursting in nonbursters, whereas raising [Ca(2+)](o) suppressed intrinsic bursting in native bursters. The induction of intrinsic bursting by low [Ca(2+)](o) was associated with enlargement of the spike ADP. Low [Ca(2+)](o)-induced intrinsic bursts and their underlying ADPs were suppressed by drugs that reduce the persistent Na(+) current (I(NaP)), indicating that this current mediates the slow burst depolarization. Blocking Ca(2+)-activated K(+) currents with extracellular Ni(2+) or intracellular chelation of Ca(2+) did not induce intrinsic bursting. This and other evidence suggest that lowering [Ca(2+)](o) may induce intrinsic bursting by augmenting I(NaP). Because repetitive neuronal activity in the hippocampus is associated with marked decreases in [Ca(2+)](o), the regulation of intrinsic bursting by extracellular Ca(2+) may provide a mechanism for preferential recruitment of this firing mode during certain forms of hippocampal activation. PMID- 11404403 TI - Increased lipid peroxidation precedes amyloid plaque formation in an animal model of Alzheimer amyloidosis. AB - Oxidative stress is a key feature in the Alzheimer's disease (AD) brain and manifests as lipid peroxidation (LPO). Isoprostanes (iPs) are specific and sensitive markers of in vivo LPO. To determine whether amyloid beta (Abeta) deposition in vivo is associated with increased LPO, we examined iP levels in a transgenic mouse model (Tg2576) of AD amyloidosis. Urine, plasma, and brain tissues were collected from Tg2576 and littermate wild-type (WT) animals at different time points starting at 4 months of age and continuing until 18 months of age. Levels of urinary 8,12-iso-iPF(2alpha)-VI were higher in Tg2576 than in WT animals as early as 8 months of age and remained this high for the rest of the study. A similar pattern was observed for plasma levels of 8,12-iso-iPF(2alpha) VI. Homogenates from the cerebral cortex and hippocampus of Tg2576 mice had higher levels of 8,12-iso-iPF(2alpha)-VI than those from WT mice starting at 8 months of age. In contrast, a surge of Abeta 1-40 and 1-42 levels as well as Abeta deposits in Tg2576 mouse brains occurred later, at 12 months of age. A direct correlation was observed between brain 8,12-iso-iPF(2alpha)-VI and Abeta 1 40 and 1-42. Because LPO precedes amyloid plaque formation in Tg2576 mice, this suggests that brain oxidative damage contributes to AD pathogenesis before Abeta accumulation in the AD brain. PMID- 11404404 TI - A labile component of AMPA receptor-mediated synaptic transmission is dependent on microtubule motors, actin, and N-ethylmaleimide-sensitive factor. AB - Glutamate receptor channels are synthesized in the cell body, are inserted into intracellular vesicles, and move to dendrites where they become incorporated into synapses. Dendrites contain abundant microtubules that have been implicated in the vesicle-mediated transport of ion channels. We have examined how the inhibition of microtubule motors affects synaptic transmission. Monoclonal antibodies that inactivate the function of dynein or kinesin were introduced into hippocampal CA1 pyramidal cells through a patch pipette. Both antibodies substantially reduced the AMPA receptor-mediated responses within 1 hr but had no effect on the NMDA receptor-mediated response. Heat-inactivated antibody or control antibodies had a much smaller effect. A component of transmission appeared to be resistant even to the combination of these inhibitors, and we therefore explored whether other agents also produce only a partial inhibition of transmission. A similar resistant component was found by using an actin inhibitor (phalloidin) or an inhibitor of NSF (N-ethylmaleimide-sensitive fusion protein)/GluR2 interaction. We then examined whether these effects were independent or occluded each other. We found that a combination of phalloidin and NSF/GluR2 inhibitor reduced the response to approximately 30% of baseline level, an effect only slightly larger than that produced by each agent alone. The addition of microtubule motor inhibitors to this combination produced no further inhibition. We conclude that there are two components of AMPA receptor-mediated transmission; one is a labile pool sensitive to NSF/GluR2 inhibitors, actin inhibitors, and microtubule motor inhibitors. A second, nonlabile pool resembles NMDA receptor channels in being nearly insensitive to any of these agents on the hour time scale of our experiments. PMID- 11404405 TI - Synapsin controls both reserve and releasable synaptic vesicle pools during neuronal activity and short-term plasticity in Aplysia. AB - Neurotransmitter release is a highly efficient secretory process exhibiting resistance to fatigue and plasticity attributable to the existence of distinct pools of synaptic vesicles (SVs), namely a readily releasable pool and a reserve pool from which vesicles can be recruited after activity. Synaptic vesicles in the reserve pool are thought to be reversibly tethered to the actin-based cytoskeleton by the synapsins, a family of synaptic vesicle-associated phosphoproteins that have been shown to play a role in the formation, maintenance, and regulation of the reserve pool of synaptic vesicles and to operate during the post-docking step of the release process. In this paper, we have investigated the physiological effects of manipulating synapsin levels in identified cholinergic synapses of Aplysia californica. When endogenous synapsin was neutralized by the injection of specific anti-synapsin antibodies, the amount of neurotransmitter released per impulse was unaffected, but marked changes in the secretory response to high-frequency stimulation were observed, including the disappearance of post-tetanic potentiation (PTP) that was substituted by post tetanic depression (PTD), and increased rate and extent of synaptic depression. Opposite changes on post-tetanic potentiation were observed when synapsin levels were increased by injecting exogenous synapsin I. Our data demonstrate that the presence of synapsin-dependent reserve vesicles allows the nerve terminal to release neurotransmitter at rates exceeding the synaptic vesicle recycling capacity and to dynamically change the efficiency of release in response to conditioning stimuli (e.g., post-tetanic potentiation). Moreover, synapsin dependent regulation of the fusion competence of synaptic vesicles appears to be crucial for sustaining neurotransmitter release during short periods at rates faster than the replenishment kinetics and maintaining synchronization of quanta in evoked release. PMID- 11404406 TI - NMDA receptor-mediated Na+ signals in spines and dendrites. AB - Spines and dendrites of central neurons represent an important site of synaptic signaling and integration. Here we identify a new, synaptically mediated spine signal with unique properties. Using two-photon Na(+) imaging, we show that suprathreshold synaptic stimulation leads to transient increases in Na(+) concentration in postsynaptic spines and their adjacent dendrites. This local signal is restricted to a dendritic domain near the site of synaptic input. In presumed active spines within this domain, the Na(+) level increases by 30-40 mm even during short bursts of synaptic stimulation. During a long-term potentiation induction protocol (100 Hz, 1 sec), the Na(+) level in the active spines reaches peak amplitudes of approximately 100 mm. We find that the Na(+) transients are mainly mediated by Na(+) entry through NMDA receptor channels and are detected during the coincident occurrence of synaptic potentials and backpropagating action potentials. The large amplitudes of the Na(+) transients and their location on dendritic spines suggest that this signal is an important determinant of electrical and biochemical spine characteristics. PMID- 11404407 TI - Drosophila alpha- and beta-spectrin mutations disrupt presynaptic neurotransmitter release. AB - Spectrins are plasma membrane-associated cytoskeletal proteins implicated in several aspects of synaptic development and function, including presynaptic vesicle tethering and postsynaptic receptor aggregation. To test these hypotheses, we characterized Drosophila mutants lacking either alpha- or beta spectrin. The Drosophila genome contains only one alpha-spectrin and one conventional beta-spectrin gene, making it an ideal system to genetically manipulate spectrin levels and examine the resulting synaptic alterations. Both spectrin proteins are strongly expressed in the Drosophila neuromusculature and highly enriched at the glutamatergic neuromuscular junction. Protein null alpha- and beta-spectrin mutants are embryonic lethal and display severely disrupted neurotransmission without altered morphological synaptogenesis. Contrary to current models, the absence of spectrins does not alter postsynaptic glutamate receptor field function or the ultrastructural localization of presynaptic vesicles. However, the subcellular localization of numerous synaptic proteins is disrupted, suggesting that the defects in presynaptic neurotransmitter release may be attributable to inappropriate assembly, transport, or localization of proteins required for synaptic function. PMID- 11404408 TI - Strain-dependent differences in calcium signaling predict excitotoxicity in murine hippocampal neurons. AB - Commonly used inbred murine strains differ substantially in their vulnerability to excitotoxic insults. We investigated whether differences in dendritic Ca(2+) signaling could underlie the differential vulnerability of C57BL/6 (resistant to kainate excitotoxicity) and C57BL/10 strains (vulnerable). A striking difference was found in fine dendrite Ca(2+) responses after kainate exposure. Ca(2+) signals in distal dendrites were large in C57BL/10 neurons, and, if a threshold concentration of approximately 1.5 microm was reached, a region of sustained high Ca(2+) was established in the distal dendritic tree. This region then served as an initiation site for a degenerative cascade, producing high Ca(2+) levels that slowly spread to involve the entire neuron and led to cell death. Dendritic Ca(2+) signals in C57BL/6 neurons were much smaller and did not trigger these propagating secondary responses. Strain differences in dendritic Ca(2+) signaling were also evident after tetanic stimulation of Schaffer collaterals. Ca(2+) responses were much larger and peaked earlier in distal dendrites of C57BL/10 compared with those in C57BL/6. Neurons from both strains had similar membrane properties and responded to kainate with intense action potential firing. Degenerative Ca(2+) responses were seen in both strains if soma Ca(2+) could be sustained above 1.5 microm. The early phases of secondary Ca(2+) responses were attributable to Ca(2+) influx and were abolished rapidly by buffered zero Ca(2+) saline. Taken together, these data indicate that the substantial difference in Ca(2+) signals in fine distal dendrites and in the initiation of spreading secondary responses may underlie the selective vulnerability of these neurons to excitotoxic insults. PMID- 11404409 TI - Ampa/kainate receptor activation mediates hypoxic oligodendrocyte death and axonal injury in cerebral white matter. AB - We developed an in situ model to investigate the hypothesis that AMPA/kainate (AMPA/KA) receptor activation contributes to hypoxic-ischemic white matter injury in the adult brain. Acute coronal brain slices, including corpus callosum, were prepared from adult mice. After exposure to transient oxygen and glucose deprivation (OGD), white matter injury was assessed by electrophysiology and immunofluorescence for oligodendrocytes and axonal neurofilaments. White matter cellular components and the stimulus-evoked compound action potential (CAP) remained stable for 12 hr after preparation. OGD for 30 min resulted in an irreversible loss of the CAP as well as structural disruption of axons and subsequent loss of neurofilament immunofluorescence. OGD also caused widespread oligodendrocyte death, demonstrated by the loss of APC labeling and the gain of pyknotic nuclear morphology and propidium iodide labeling. Blockade of AMPA/KA receptors with 30 microm NBQX or the AMPA-selective antagonist 30 microm GYKI 52466 prevented OGD-induced oligodendrocyte death. Oligodendrocytes also were preserved by the removal of Ca(2+), but not by a blockade of voltage-gated Na(+) channels. The protective action of NBQX was still present in isolated corpus callosum slices. CAP areas and axonal structure were preserved by Ca(2+) removal and partially protected by a blockade of voltage-gated Na(+) channels. NBQX prevented OGD-induced CAP loss and preserved axonal structure. These observations highlight convergent pathways leading to hypoxic-ischemic damage of cerebral white matter. In accordance with previous suggestions, the activation of voltage gated Na(+) channels contributes to axonal damage. Overactivation of glial AMPA/KA receptors leads to oligodendrocyte death and also plays an important role in structural and functional disruption of axons. PMID- 11404410 TI - BDNF enhances quantal neurotransmitter release and increases the number of docked vesicles at the active zones of hippocampal excitatory synapses. AB - Brain-derived neurotrophic factor (BDNF) is emerging as a key mediator of activity-dependent modifications of synaptic strength in the CNS. We investigated the hypothesis that BDNF enhances quantal neurotransmitter release by modulating the distribution of synaptic vesicles within presynaptic terminals using organotypic slice cultures of postnatal rat hippocampus. BDNF specifically increased the number of docked vesicles at the active zone of excitatory synapses on CA1 dendritic spines, with only a small increase in active zone size. In agreement with the hypothesis that an increased docked vesicle density enhances quantal neurotransmitter release, BDNF increased the frequency, but not the amplitude, of AMPA receptor-mediated miniature EPSCs (mEPSCs) recorded from CA1 pyramidal neurons in hippocampal slices. Synapse number, independently estimated from dendritic spine density and electron microscopy measurements, was also increased after BDNF treatment, indicating that the actions of BNDF on mEPSC frequency can be partially attributed to an increased synaptic density. Our results further suggest that all these actions were mediated via tyrosine kinase B (TrkB) receptor activation, established by inhibition of plasma membrane tyrosine kinases with K-252a. These results provide additional evidence of a fundamental role of the BDNF-TrkB signaling cascade in synaptic transmission, as well as in cellular models of hippocampus-dependent learning and memory. PMID- 11404411 TI - p27Kip1 and p57Kip2 regulate proliferation in distinct retinal progenitor cell populations. AB - In the developing vertebrate retina, progenitor cell proliferation must be precisely regulated to ensure appropriate formation of the mature tissue. Cyclin kinase inhibitors have been implicated as important regulators of proliferation during development by blocking the activity of cyclin-cyclin-dependent kinase complexes. We have found that the p27(Kip1) cyclin kinase inhibitor regulates progenitor cell proliferation throughout retinal histogenesis. p27(Kip1) is upregulated during the late G(2)/early G(1) phase of the cell cycle in retinal progenitor cells, where it interacts with the major retinal D-type cyclin-cyclin D1. Mice deficient for p27(Kip1) exhibited an increase in the proportion of mitotic cells throughout development as well as extensive apoptosis, particularly during the later stages of retinal histogenesis. Retroviral-mediated overexpression of p27(Kip1) in mitotic retinal progenitor cells led to premature cell cycle exit yet had no dramatic effects on Muller glial or bipolar cell fate specification as seen with the Xenopus cyclin kinase inhibitor, p27(Xic1). Consistent with the overexpression of p27(Kip1), mice lacking one or both alleles of p27(Kip1) maintained the same relative ratios of each major retinal cell type as their wild-type littermates. During the embryonic stages of development, when both p27(Kip1) and p57(Kip2) are expressed in retinal progenitor cells, they were found in distinct populations, demonstrating directly that different retinal progenitor cells are heterogeneous with respect to their expression of cell cycle regulators. PMID- 11404412 TI - A role of netrin-1 in the formation of the subcortical structure striatum: repulsive action on the migration of late-born striatal neurons. AB - The mammalian striatum arises in the basal telencephalon and contains morphologically homogenous neurons that can be divided into two distinct compartments, patches and the matrix. During development, patch neurons are generated first to form a striatal primordium. After a large influx of later-born matrix neurons into this region, the unique mosaic arrangement of these two neuronal phenotypes is established. The massive migration of matrix neurons continues during the embryonic period, and they eventually comprise 80-85% of the mature striatum. To elucidate the cellular mechanism or mechanisms underlying this critical event in striatal histogenesis, we examined the migration characteristics of striatal subventricular zone (SVZ) cells at embryonic day 18 when neurogenesis peaks for matrix neurons. Using gel cultures, we show that netrin-1, one of the diffusible guidance cues expressed in the striatal ventricular zone (VZ), exerts a repulsive action on migrating SVZ cells. This effect is blocked in the presence of antibodies against Deleted in colorectal cancer (DCC), a putative receptor for netrin-1. The expression patterns of netrin 1 and DCC strongly suggest the involvement of this effect in the outward migration of SVZ cells into the striatal postmitotic region. Our cell tracing study using living brain slices demonstrates that striatal SVZ cells migrate toward and disperse throughout the striatum, in which they differentiate into phenotypes of striatal projection neurons. We suggest that netrin-1 expressed in the striatal VZ serves to guide the large influx of striatal matrix neurons into the striatal primordium and is thereby involved in the initial formation of fundamental striatal structures. PMID- 11404413 TI - Diversity and specificity of actions of Slit2 proteolytic fragments in axon guidance. AB - The Slits are secreted proteins that bind to Robo receptors and play a role in axon guidance and neuronal migration. In vertebrates, Slit2 is a major chemorepellent for developing axons and is involved in the control of midline crossing. In vivo, Slit2 is cleaved into 140 kDa N-terminal (Slit2-N) and 55-60 kDa C-terminal (Slit2-C) fragments, although the uncleaved/full-length form can also be isolated from brain extract. We explored the functional activities of Slit2 fragments by engineering mutant and truncated versions of Slit2 representing the N-, C-, and full/uncleavable (Slit2-U) fragments. Only Slit2-N and Slit2-U bind the Robo proteins. We found that in collagen gel, olfactory bulb (OB) but not dorsal root ganglia (DRG) axons are repelled by Slit2-N and Slit2-U. Moreover, only Slit2-N membranes or purified protein-induced OB growth cones collapse. Finally, we found that only recombinant Slit2-N could induce branching of DRG axons and that this effect was antagonized by Slit2-U. Therefore, different axons have distinct responses to Slit2 fragments, and these proteins have different growth-promoting capacities. PMID- 11404414 TI - Repellent signaling by Slit requires the leucine-rich repeats. AB - Slit is a repellent axon guidance cue produced by the midline glia in Drosophila that is required to regulate the formation of contralateral projections and the lateral position of longitudinal tracts. Four sequence motifs comprise the structure of Slit: a leucine-rich repeat (LRR), epidermal growth factor-like (EGF) repeats, a laminin-like globular (G)-domain, and a cysteine domain. Here we demonstrate that the LRR is required for repellent signaling and in vitro binding to Robo. Repellent signaling by slit is reduced by point mutations that encode single amino acid changes in the LRR domain. By contrast to the EGF or G-domains, the LRR domain is required in transgenes to affect axon guidance. Finally, we show that the midline repellent receptor, Robo, binds Slit proteins with internal deletions that also retain repellent activity. However, Robo does not bind Slit protein missing the LRR. Taken together, our data demonstrate that Robo binding and repellent signaling by Slit require the LRR region. PMID- 11404415 TI - Suppression of cortical NMDA receptor function prevents development of orientation selectivity in the primary visual cortex. AB - Selectivity to visual stimulus orientation is a basic cortical functional property believed to be crucial for normal vision. Maturation of this neuronal property requires neural activity. Still, it is unclear what might be the molecular basis for such activity-dependent processes and whether activity has an instructive or permissive role in development of orientation selectivity. There is strong evidence that the NMDA subtype of the glutamate receptor regulates activity-dependent mechanisms of ocular dominance plasticity during cortical development. For this reason, we have hypothesized that the NMDA receptor participates in activity-dependent mechanisms that sculpt orientation selectivity of cortical neurons. We used chronic in vivo infusion of antisense oligodeoxynucleotides (ODNs) to suppress NMDA receptor function in primary visual cortex during the period when orientation selectivity develops in ferrets. Chronic suppression of NMDA receptor function prevented the development of orientation and stimulus size selectivity in most cortical cells tested. In contrast, treatment with control sense or missense ODNs did not affect development of orientation selectivity, indicating specificity of effects. Importantly, antisense ODN treatment did not impair visually driven activity, which is required for development to occur. Moreover, orientation selectivity of cortical cells was not disrupted by antisense ODN treatment in mature animals, indicating developmental relevance of the effects. In conclusion, our findings document for the first time that cortical NMDA receptors are essential for the maturation of orientation selectivity. This result supports the notion that activity has an instructive role in sculpting the connections that underlie orientation selectivity in visual cortex. PMID- 11404416 TI - Unique functional properties of on and off pathways in the developing mammalian retina. AB - In the mature retina, the dendrites of On and Off ganglion cells are segregated into separate sublaminas of the inner plexiform layer, but early in development these processes are multistratified, ramifying more widely within this synaptic layer. The dendritic pattern exhibited by immature ganglion cells suggests that there may be a functional convergence of On and Off pathways in the developing retina, but previous studies have provided evidence against this. Here we demonstrate by patch-clamp recordings and dye filling that ganglion cells with multistratified dendrites respond to the onset, as well as the offset, of light. We further show that, in the dark-adapted retina, the glutamate analog 2-amino-4 phosphonobutric acid abolishes On and Off discharges in ganglion cells with multistratified dendrites. In contrast, in cells with stratified dendrites, this drug selectively blocks On responses. These findings provide evidence for unique functional attributes of On and Off pathways in the developing retina. The properties of immature ganglion cells documented here have important implications for the roles ascribed to neuronal activity in refining connections during the early development of the visual system. PMID- 11404417 TI - The role of nitric oxide in development of topographic precision in the retinotectal projection of chick. AB - The axonal projection from the retina to the tectum exhibits a precise topographic order in the mature chick such that neighboring ganglion cells send axons to neighboring termination zones in the contralateral tectum. The initial pattern formed during development is much less organized and is refined to the adult pattern during a discrete period of development. Refinement includes elimination of radically aberrant projections, such as those from the temporal side of the retina to posterior regions of the tectum, as well as a more subtle improvement in the topographic precision of the projection. The enzyme that synthesizes nitric oxide is expressed at high levels in the tectum during the developmental period in which the topography improves. Pharmacological blockade of nitric oxide synthesis during this period prevented elimination of topographically inappropriate retinotectal projections in a dose-dependent manner. This effect could not be duplicated by treatment of embryos with a vasoconstrictor, indicating that vascular changes were not a factor. These results show that nitric oxide is involved in refinement of the topography of the retinotectal projection as well as in other aspects of refinement of this projection in developing chick. PMID- 11404419 TI - Induction of the plasminogen activator system accompanies peripheral nerve regeneration after sciatic nerve crush. AB - Peripheral nerve regeneration is dependent on the ability of regenerating neurites to migrate through cellular debris and altered extracellular matrix at the injury site, grow along the residual distal nerve sheath conduit, and reinnervate synaptic targets. In cell culture, growth cones of regenerating axons secrete proteases, specifically plasminogen activators (PAs), which are believed to facilitate growth cone movement by digesting extracellular matrices and cell adhesions. In this study, the PA system was shown to be specifically activated in sensory neurons after sciatic nerve crush in adult mice. The number of sensory neurons expressing urokinase PA receptor (uPAR) mRNA levels increased above sham levels by 8 hr after crush, whereas the number of sensory neurons expressing uPA and tissue PA (tPA) mRNAs was significantly increased by 3 d after crush. PA mRNA levels were also increased at the crush site, with uPA mRNA elevated by 8 hr after crush and tPA and uPAR mRNA levels markedly increased by 7 d. PA-dependent enzymatic activity was significantly increased from 1 to 7 d after crush in nerves that had been crushed compared with uncrushed nerves. Immunohistochemistry showed that tPA was localized within regenerating axons of the sciatic nerve. There were no significant changes in plasminogen activator inhibitor 1 activity between crush and sham after the injury. These results clearly demonstrated that after injury the PA system was rapidly induced in sensory neurons, where it may play an important role in nerve regeneration in vivo. PMID- 11404418 TI - Sonic hedgehog facilitates dopamine differentiation in the presence of a mesencephalic glial cell line. AB - The aim of this study was to establish a cellular system to investigate the requirement for cell surface and diffusible molecules in the differentiation of fetal mesencephalic cells toward the dopamine lineage. Toward this end, we immortalized rat embryonic day 14 (E14) mesencephalon with a regulatable retroviral vector encoding v-myc. The stably transduced cells were pooled and designated as VME14 cells. VME14 cells proliferated rapidly, stopped proliferating, extended processes, and expressed GFAP after suppression of the v myc expression with tetracycline, suggesting that VME14 cells differentiated into glial cells. Dissociated cells derived from the E11 rat mesencephalon gave rise to only a small number of tyrosine hydroxylase (TH)-positive neurons. However, when grown on a monolayer of the differentiated VME14 cells, a significantly higher number of cells differentiated into TH-positive neurons. VME14 cells were transduced with the secreted N-terminal cleavage product of the Sonic hedgehog gene (SHH-N), an inducer of mesencephalic dopaminergic neurons. This monoclonal, SHH-N-overexpressing cell line further enhanced dopaminergic differentiation of E11 rat mesencephalon cells. Thus, SHH-N and signals derived from fetal mesencephalic glia act cooperatively to facilitate dopaminergic differentiation. These fetal mesencephalon-derived cell lines will provide tools for the study of signals involved in dopaminergic differentiation. PMID- 11404420 TI - Mice lacking tPA, uPA, or plasminogen genes showed delayed functional recovery after sciatic nerve crush. AB - Axonal outgrowth during peripheral nerve regeneration relies on the ability of growth cones to traverse through an environment that has been altered structurally and along a basal lamina sheath to reinnervate synaptic targets. To promote migration, growth cones secrete proteases that are thought to dissolve cell-cell and cell-matrix adhesions. These proteases include the plasminogen activators (PAs), tissue PA (tPA) and urokinase PA (uPA), and their substrate, plasminogen. PA expression and secretion are upregulated in regenerating mammalian sensory neurons in culture. After sciatic nerve crush in mice, there was an induction of PA mRNAs in the sensory neurons contributing to the crushed nerve and an upregulation of PA-dependent activity in crushed nerve compared with sham counterparts during nerve regeneration. To further assess the role of the PA system during peripheral nerve regeneration, PA-dependent activity as well as recovery of sensory and motor function in the injured hindlimb were assessed in wild-type, tPA, uPA, and plasminogen knock-out mice. Protease activity visualized by gel zymography showed that after nerve crush, the upregulation of PA activity in the tPA and uPA knock-out mice was delayed compared with wild-type mice. Recovery of sensory function was assessed by toe pinch, footpad prick, and the toe-spreading reflex. All knock-out mice demonstrated a significant delay in hindlimb response to these sensory stimuli compared with wild-type mice. For each modality tested, the uPA knock-out mice were the most dramatically affected, showing the longest delay to initiate a response. These studies clearly showed that PAs were necessary for timely functional recovery by regenerating peripheral nerves. PMID- 11404421 TI - Gabaergic inhibition antagonizes adaptive adjustment of the owl's auditory space map during the initial phase of plasticity. AB - We studied the influence of GABA-mediated inhibition on adaptive adjustment of the owl's auditory space map during the initial phase of plasticity. Plasticity of the auditory space map was induced by subjecting owls to a chronic prismatic displacement of the visual field. In the initial stages of plasticity, inhibition suppressed responses to behaviorally appropriate, newly functional excitatory inputs. As a result, adaptive changes in excitatory input were only partially expressed as postsynaptic spike activity. This masking effect of inhibition on map plasticity did not depend on the activity of NMDA receptors at the synapses that supported the newly learned responses. On the basis of these results, we propose that the pattern of feedforward inhibition is less dynamic than the pattern of feedforward excitation at the site of plasticity. As a result, initially in the adjustment process the preexisting pattern of feedforward GABAergic inhibition opposes changes in the auditory space map and tends to preserve the established response properties of the network. The implications of this novel role of inhibition for the functional plasticity of the brain are discussed. PMID- 11404422 TI - Blockade of endogenous neurotrophic factors prevents the androgenic rescue of rat spinal motoneurons. AB - Target-derived neurotrophic factors are assumed to regulate motoneuron cell death during development but remain unspecified. Motoneuron cell death in the spinal nucleus of the bulbocavernosus (SNB) of rats extends postnatally and is controlled by androgens. We exploited these features of the SNB system to identify endogenously produced trophic factors regulating motoneuron survival. Newborn female rat pups were treated with the androgen, testosterone propionate, or the oil vehicle alone. In addition, females received trophic factor antagonists delivered either into the perineum (the site of SNB target muscles) or systemically. Fusion molecules that bind and sequester the neurotrophins (trkA IgG, trkB-IgG, and trkC-IgG) were used to block activation of neurotrophin receptors, and AADH-CNTF was used to antagonize signaling through the ciliary neurotrophic factor receptor-alpha (CNTFRalpha). An acute blockade of trkB, trkC, or CNTFRalpha prevented the androgenic sparing of SNB motoneurons when antagonists were delivered to the perineum. Trophic factor antagonists did not significantly reduce SNB motoneuron number when higher doses were injected systemically. These findings demonstrate a requirement for specific, endogenously produced trophic factors in the androgenic rescue of SNB motoneurons and further suggest that trophic factor interactions at the perineum play a crucial role in masculinization of this neural system. PMID- 11404423 TI - Neuronal expression of synaptotagmin-related gene 1 is regulated by thyroid hormone during cerebellar development. AB - Thyroid hormone (TH) is essential for proper brain development, acting through nuclear receptors that modulate the expression of specific genes in response to hormone binding. In a screen for genes regulated by TH in the rat cerebellum, we recently identified a novel gene, synaptotagmin-related gene 1 (Srg1). The Srg1 protein is structurally similar to synaptotagmins, a family of proteins involved in regulating neurotransmission. To elucidate a potential role of Srg1 in brain development, we have investigated the developmental and TH-regulated expression of Srg1 in the neonatal rat brain. We show that expression of both Srg1 RNA and protein is detected only in the brain and specifically in neurons. Srg1 mRNA and protein levels increase postnatally, nearing adult levels after the third postnatal week. Neonatal TH deficiency results in a significant reduction and delay in expression of both Srg1 RNA and protein. Using immunohistochemistry, we were able to detect Srg1 protein in numerous brain regions. In the cerebellum, Srg1 protein is localized to the molecular layer, indicating that it is highly expressed in granule cell axons. To further examine Srg1 expression in cerebellar granule cells (CGCs), we used an in vitro cell culture model. In primary cultures of CGCs, Srg1 expression is significantly reduced in the absence of TH. Srg1 mRNA is rapidly upregulated in cultured CGCs, suggesting a direct response to TH. Neuronal and TH-regulated expression of Srg1, together with its localization to neurites, implicates Srg1 as an important component of the program of gene expression induced by TH in the developing brain. PMID- 11404424 TI - Chronic prenatal ethanol exposure increases GABA(A) receptor subunit protein expression in the adult guinea pig cerebral cortex. AB - Excessive consumption of ethanol during pregnancy can produce teratogenic effects in offspring and is the leading cause of mental deficiency in the Western world. The objective of this study was to examine the effects of chronic prenatal ethanol exposure on the number of GABA(A) receptors and relative protein levels for GABA(A) receptor alpha1 and beta2/3 subunits in the adult guinea pig cerebral cortex. Timed pregnant Dunkin-Hartley strain guinea pigs were given one of the following oral treatments daily throughout gestation: 4 gm of ethanol per kilogram of maternal body weight, isocaloric-sucrose with pair feeding, or isovolumetric water with ad libitum access to food. The ethanol treatment resulted in a peak maternal blood ethanol concentration of 328 +/- 55 mg/dl (71.3 +/- 12.0 mm) on gestational day 57 (term, approximately 68 d). Chronic prenatal exposure to ethanol resulted in increased spontaneous locomotor activity throughout development and decreased cerebral cortical weight in adult offspring. The number of cerebral cortical [(3)H]muscimol binding sites was increased in adult offspring from the ethanol treatment group, and there was a corresponding increase in the amount of GABA(A) receptor alpha1 and beta2/3 subunit proteins in these same animals. For individual offspring, there were correlations between locomotor activity and cerebral cortical weight, as well as between cerebral cortical weight and GABA(A) receptor neurochemistry. There was no effect of chronic prenatal ethanol exposure on [(3)H]MK-801 binding in this tissue. These data demonstrate that chronic prenatal ethanol exposure has long-term consequences on the regulation of GABA(A) receptor expression in the cerebral cortex. PMID- 11404425 TI - Galpha(olf) levels are regulated by receptor usage and control dopamine and adenosine action in the striatum. AB - In the striatum, dopamine D(1) and adenosine A(2A) receptors stimulate the production of cAMP, which is involved in neuromodulation and long-lasting changes in gene expression and synaptic function. Positive coupling of receptors to adenylyl cyclase can be mediated through the ubiquitous GTP-binding protein Galpha(S) subunit or through the olfactory isoform, Galpha(olf), which predominates in the striatum. In this study, using double in situ hybridization, we show that virtually all striatal efferent neurons, identified by the expression of preproenkephalin A, substance P, or D(1) receptor mRNA, contained high amounts of Galpha(olf) mRNA and undetectable levels of Galpha(s) mRNA. In contrast, the large cholinergic interneurons contained both Galpha(olf) and Galpha(s) transcripts. To assess the functional relationship between dopamine or adenosine receptors and G-proteins, we examined G-protein levels in the striatum of D(1) and A(2A) receptor knock-out mice. A selective increase in Galpha(olf) protein was observed in these animals, without change in mRNA levels. Conversely, Galpha(olf) levels were decreased in animals lacking a functional dopamine transporter. These results indicate that Galpha(olf) protein levels are regulated through D(1) and A(2A) receptor usage. To determine the functional consequences of changes in Galpha(olf) levels, we used heterozygous Galpha(olf) knock-out mice, which possess half of the normal Galpha(olf) levels. In these animals, the locomotor effects of amphetamine and caffeine, two psychostimulant drugs that affect dopamine and adenosine signaling, respectively, were markedly reduced. Together, these results identify Galpha(olf) as a critical and regulated component of both dopamine and adenosine signaling. PMID- 11404426 TI - Maturation of extinction behavior in infant rats: large-scale regional interactions with medial prefrontal cortex, orbitofrontal cortex, and anterior cingulate cortex. AB - The ability to express a behavior during the postnatal period may be related to developmental changes in the recruitment of particular neural systems. Here, we show that developmental changes in the functional interactions involving three cortical regions (the medial prefrontal cortex, orbitofrontal cortex, and anterior cingulate cortex) are associated with maturation of extinction behavior in infant rats. Postnatal day 17 (P17) and P12 pups were trained in a straight alley runway on an alternating schedule of reward and nonreward [patterned single alternation (PSA)] or on a pseudorandom schedule of partial reinforcement (PRF); the pups were then injected with fluorodeoxyglucose (FDG) and shifted to continuous nonreward (extinction). Handled control groups exposed to the same training environment but not trained on a particular schedule were included. Among P17 pups, extinction proceeded faster in PSA pups relative to PRF pups. No differences were found between P12 groups. FDG uptake, an index of acute changes in functional activity, was quantified in the three cortical regions and 27 other brain regions of interest. A multivariate covariance analysis, seed partial least squares, revealed that functional relationships involving the three cortical regions and large-scale systems of regions throughout the rostrocaudal extent of the brain changed with training in P17 pups. The cortical regions were primarily uncoupled in the younger group. The data suggest that functional maturation of the frontal cortical regions and their interactions with other brain systems are related to the maturational shift in behavior. PMID- 11404427 TI - Auditory space-time receptive field dynamics revealed by spherical white-noise analysis. AB - Numerous studies have investigated the spatial sensitivity of cat auditory cortical neurons, but possible dynamic properties of the spatial receptive fields have been largely ignored. Given the considerable amount of evidence that implicates the primary auditory field in the neural pathways responsible for the perception of sound source location, a logical extension to earlier observations of spectrotemporal receptive fields, which characterize the dynamics of frequency tuning, is a description that uses sound source direction, rather than sound frequency, to examine the evolution of spatial tuning over time. The object of this study was to describe auditory space-time receptive field dynamics using a new method based on cross-correlational techniques and white-noise analysis in spherical auditory space. This resulted in a characterization of auditory receptive fields in two spherical dimensions of space (azimuth and elevation) plus a third dimension of time. Further analysis has revealed that spatial receptive fields of neurons in auditory cortex, like those in the visual system, are not static but can exhibit marked temporal dynamics. This might result, for example, in a neuron becoming selective for the direction and speed of moving auditory sound sources. Our results show that approximately 14% of AI neurons evidence significant space-time interaction (inseparability). PMID- 11404428 TI - Oriented axon projections in primary visual cortex of the monkey. AB - One important aspect of the functional architecture of primary visual cortex is the circuitry that accounts for the receptive field properties of neurons. The anatomy that underlies retinotopy and ocular dominance is well known, but no anatomical structure related to orientation selectivity has been found in primates. We examined whether the arrangement of local axon systems projecting within the cortical layers might be correlated with orientation preference in New World monkeys. We found that axons in layer 3 spread out from the site of a tracer injection in an anisotropic manner and that this elongated distribution is aligned with the preferred orientation recorded at each site. Moreover, within a few degrees of the foveal representation, the majority of the axon terminals fall within or just outside of the limits of the cortical mapping of the classical receptive field. Thus local axons produce a field of monosynaptic excitation that aligns with orientation axes and reaches neurons that have receptive fields which are adjacent in visual space. PMID- 11404429 TI - Forced limb-use effects on the behavioral and neurochemical effects of 6 hydroxydopamine. AB - Rats with unilateral depletion of striatal dopamine (DA) show marked preferential use of the ipsilateral forelimb. Previous studies have shown that implementation of motor therapy after stroke improves functional outcome (Taub et al., 1999). Thus, we have examined the impact of forced use of the impaired forelimb during or soon after unilateral exposure to the DA neurotoxin 6-hydroxydopamine (6 OHDA). In one group of animals, the nonimpaired forelimb was immobilized using a cast, which forced exclusive use of the impaired limb for the first 7 d after infusion. The animals that received a cast displayed no detectable impairment or asymmetry of limb use, could use the contralateral (impaired) forelimb independently for vertical and lateral weight shifting, and showed no contralateral turning to apomorphine. The behavioral effects were maintained throughout the 60 d of observation. In addition to the behavioral sparing, these animals showed remarkable sparing of striatal DA, its metabolites, and the expression of the vesicular monoamine transporter, suggesting a decrease in the extent of DA neuron degeneration. Behavioral and neurochemical sparing appeared to be complete when the 7 d period of immobilization was initiated immediately after 6-OHDA infusion, only partial sparing was evident when immobilization was initiated 3 d postoperatively, and no sparing was detected when immobilization was initiated 7 d after 6-OHDA treatment. These results suggest that physical therapy may be beneficial in Parkinson's disease. PMID- 11404431 TI - Dorsal hippocampal kindling produces a selective and enduring disruption of hippocampally mediated behavior. AB - Kindling produces enduring neural changes that are subsequently manifest in enhanced susceptibility to seizure-evoking stimuli and alterations in some types of behavior. The present study investigated the effects of dorsal hippocampal (dHPC) kindling on a variety of behaviors to clarify the nature of previously reported effects on spatial task performance. Rats were kindled twice daily with dHPC stimulation until three fully generalized seizures were evoked. Beginning 7 d later and on successive days, rats were tested in an elevated plus maze, a large circular open field, an open field object exploration task, and a delayed match-to-place (DMTP) task in a water maze to assess anxiety-related and activity related behavior (tasks 1 and 2), object recognition memory (task 3), and spatial cognition (task 4). Kindling disrupted performance on the DMTP task in a manner that was not delay dependent and produced a mild enhancement of activity-related behaviors in the open field task but not the elevated plus maze. All other aspects of testing were spared. These findings indicate that dHPC kindling produces enduring and selective effects on behavior that are consistent with a restricted disruption of hippocampally mediated functions. Possible bases for these effects are changes in local NMDA receptor function and/or changes in local inhibition, which might alter the optimal conditions for experience-dependent induction of intrahippocampal plasticity. This preparation may be useful for studying the mechanisms of mnemonic dysfunction associated with temporal lobe epilepsy and may offer unique insights into the mechanisms underlying normal hippocampal function. PMID- 11404430 TI - Endomorphin-1: induction of motor behavior and lack of receptor desensitization. AB - The endomorphins are recently discovered endogenous agonists for the mu-opioid receptor (Zadina et al., 1997). Endomorphins produce analgesia; however, their role in other brain functions has not been elucidated. We have investigated the behavioral effects of endomorphin-1 in the globus pallidus, a brain region that is rich in mu-opioid receptors and involved in motor control. Bilateral administration of endomorphin-1 in the globus pallidus of rats induced orofacial dyskinesia. This effect was dose-dependent and at the highest dose tested (18 pmol per side) was sustained during the 60 min of observation, indicating that endomorphin-1 does not induce rapid desensitization of this motor response. In agreement with a lack of desensitization of mu-opioid receptors, 3 hr of continuous exposure of the cloned mu receptor to endomorphin-1 did not diminish the subsequent ability of the agonist to inhibit adenylate cyclase activity in cells expressing the cloned mu-opioid receptor. Confirming the involvement of mu opioid receptors, the behavioral effect of endomorphin-1 in the globus pallidus was blocked by the opioid antagonist naloxone and the mu-selective peptide antagonist Cys(2)-Tyr(3)-Orn(5)-Pen(7) amide (CTOP). Furthermore, the selective mu receptor agonist [d-Ala(2)-N-Me-Phe(4)-Glycol(5)]-enkephalin (DAMGO) also stimulated orofacial dyskinesia when infused into the globus pallidus, albeit transiently. Our findings suggest that endogenous mu agonists may play a role in hyperkinetic movement disorders by inducing sustained activation of pallidal opioid receptors. PMID- 11404432 TI - Morphine-induced dependence and sensitization are altered in mice deficient in AMPA-type glutamate receptor-A subunits. AB - AMPA-type glutamate receptors have been suggested to be involved in the neurobiological mechanisms of drug addiction. We have made use of two mouse lines, which both have modulated AMPA receptor responses. The first line is entirely deficient in glutamate receptor-A (GluR-A) subunits (A-/- knock-out line) and, in the second one, the Q582 residue of GluR-A subunits is replaced by an arginine residue (R/R mutants), which reduces the calcium permeability and channel conductance of the receptors containing this mutated subunit. Mice of both lines are healthy, but they show slightly increased locomotor activity. Acute morphine administration enhanced locomotor activity of the GluR-A-/- and GluR-A(R/R) mice, at least as much as that of their wild-type littermates. Only in the GluR-A-/- mice did we observe reduced tolerance development in tail-flick antinociception and less severe naloxone-precipitated withdrawal symptoms after treatment with increasing morphine doses, without differences in plasma and brain morphine levels when compared with wild type. Repeated daily morphine administration sensitized the locomotor activity responses in the GluR-A-/- and GluR-A(R/R) mice only when given in the measuring cages, whereas the wild-type mice showed slightly increased responses also when the repeated treatment was given in their home cages. Normal or even enhanced context-dependent sensitization was observed also with repeated amphetamine administration in the GluR-A subunit-deficient mice. The results indicate that AMPA receptors are involved in the acute and chronic effects of morphine, including context independent sensitization, and that the GluR-A subunit itself is important for morphine tolerance and dependence. PMID- 11404433 TI - Capsaicin responses in heat-sensitive and heat-insensitive A-fiber nociceptors. AB - The recently cloned vanilloid receptor (VR1) is postulated to account for heat and capsaicin sensitivity in unmyelinated afferents. We sought to determine whether heat and capsaicin sensitivity also coexist in myelinated nociceptive afferents. Action potential (AP) activity was recorded from single A-fiber nociceptors that innervated the hairy skin in monkey. Before intradermal injection of capsaicin (10 microg/10 microl) into the receptive field, nociceptors were classified as heat-sensitive (threshold, 1 hr. Combining HFS with feedback from the lid movement was critical for this facilitation because presenting HFS immediately after the blink did not alter subsequent blinks. When HFS preceded the blink, however, this treatment suppressed subsequent blinks for 30 min. These effects appear to occur within the trigeminal reflex blink circuits rather than at motoneurons, because stimulation of the previously HFS-treated SO evoked altered blinks in both eyelids, whereas stimulation of the untreated SO elicited unaltered blinks in both eyelids. The modified blink amplitude resulted from altering the response to A-fiber inputs to the trigeminal nerve because all stimuli were too weak to activate C-fibers. The data suggest that HFS produce LTP- and long-term depression (LTD)-like effects on wide dynamic range neurons in the trigeminal reflex blink circuit. The data also support the hypothesis that LTP and LTD mechanisms play a role in adaptive modification of human reflex blinks. PMID- 11404444 TI - Getting Occupational and Environmental Medicine to developing countries. PMID- 11404445 TI - Occupational health research priorities in Malaysia: a Delphi study. AB - OBJECTIVES: As part of a consultancy project on occupational health, the Delphi method was used to identify research priorities in occupational health in Malaysia. METHODS: Participation was sought from government ministries, industry, and professional organisations, and university departments with an interest in occupational and public health. Two rounds of questionnaires resulted in a final list of priorities, with noticeable differences between participants depending on whether they worked in industry or were from government organisations. RESULTS: The participation rate of 71% (55 of 78) was obtained for the first questionnaire and 76% (72 of 95) for the second questionnaire. The participants identified occupational health problems for specific groups and industries as the top research priority area (ranked as top priority by 25% of participants). Ministry of Health participants placed emphasis on healthcare workers (52% ranking it as top priority), whereas those from industry identified construction and plantation workers as groups, which should be accorded the highest priority. Evaluation of research and services was given a low priority. CONCLUSIONS: The priorities for occupational health determined with the Delphi approach showed differences between Malaysia, a developing country, and findings from similar European studies. This may be expected, as differences exist in stages of economic development, types of industries, occupational activities, and cultural attitudes to occupational health and safety. Chemical poisonings and workplace accidents were accorded a high priority. By contrast with findings from western countries, workplace psychosocial problems and musculoskeletal injuries were deemed less important. There also seemed to be greater emphasis on adopting interventions for identified problems based on experience in other countries rather than the need to evaluate local occupational health provisions. PMID- 11404446 TI - Sickness absence in diabetic employees. AB - OBJECTIVES: To compare sickness absence among diabetic and non-diabetic employees. METHODS: A cross sectional case-control study was conducted in a random sample of 400 diabetic employees 21--50 years old from Ljubljana that compared their sickness absence in the year 1996 with sickness absence of non diabetic employees matched by sex, age, and occupation. Sickness absence was compared in total and also in subgroups formed by sex, age, occupation, and disability. Non-parametric statistics were used (chi(2) test, Wilcoxon matched pairs test). RESULTS: The randomised sample consisted of 61.2% of men (245) and 38.8% of women (155) with a mean age of 42.5 years. Unskilled workers made up 30.2% of employees, and less than 16.4% were highly educated. Among diabetic employees there were 7.0% disabled and among non-diabetic employees 2.0%. The mean frequency of sickness absences of diabetic employees was 0.89 times in the year 1996 (95% confidence interval (95% CI) 0.70 to 1.08), and of non-diabetic employees 0.56 times (95% CI 0.47 to 0.65), p=0.01. The mean total duration of sickness absence of diabetic employees was 31.71 days (95% CI 24.86 to 38.57), of non-diabetic employees 16.57 days (95% CI 11.72 to 21.42), p<0.01. Differences were also found in subgroups but the size of subgroups was not sufficient to detect significant differences. CONCLUSIONS: The study confirmed that diabetes affects the ability to work. Appropriate work and good control of the disease are important to prevent long term complications. PMID- 11404447 TI - Foetal growth and duration of gestation relative to water chlorination. AB - OBJECTIVE: To assess the effect of exposure to chlorination byproducts during pregnancy on foetal growth and duration of pregnancy. METHODS: A population based study was conducted of 137,145 Norwegian children born alive in 1993--5. Information was obtained from the Norwegian medical birth registry, waterwork registry, and social science data service. The outcomes of interest were birth weight, low birth weight (<2500 g), small for gestational age, and preterm delivery (gestational age <37 weeks). The exposure assessment was based on quality of drinking water in the municipality where the mother lived during pregnancy. Municipal exposure was calculated with information on chlorination and the amount of natural organic matter in raw water measured as colour in mg precipitate/l. The main exposure category was high colour and chlorination, which was contrasted with the reference category of low colour and no chlorination. RESULTS: In logistic regression analysis adjusting for confounding, the risks of low birth weight (odds ratio (OR) 0.97, 95% confidence interval (95% CI) 0.89 to 1.06) and small for gestational age (OR 1.00, 95% CI 0.91 to 1.10) were not related to exposure. Contrary to the hypothesis, the risk of preterm delivery was slightly lower among the exposed than the reference category (OR 0.91, 95% CI 0.84 to 0.99). The risks of the studied outcomes were similar in newborn infants exposed to high colour drinking water without chlorination and chlorinated drinking water with low colour compared with the reference category. CONCLUSIONS: The present study did not provide evidence that prenatal exposure to chlorination byproducts at the relatively low concentrations encountered in Norwegian drinking water increases the risk of the studied outcomes. PMID- 11404448 TI - Relation between trihalomethane compounds and birth defects. AB - OBJECTIVES: To evaluate the risk of birth defects relative to exposure to specific trihalomethanes in public water supplies. METHODS: A retrospective cohort study was conducted based on data from a population based perinatal database in Nova Scotia, Canada and from the results of routine water monitoring tests. The cohort consisted of women who had a singleton birth in Nova Scotia between 1988 and 1995 and who lived in an area with a municipal water supply. The birth defects analyzed included neural tube defects, cardiovascular defects, cleft defects, and chromosomal abnormalities. Two of the four trihalomethane compounds occur in large enough concentrations to be analyzed (chloroform and bromodichloromethane (BDCM)). RESULTS: Exposure to BDCM at concentrations of 20 microg/l or over was associated with an increased risk of neural tube defects (adjusted relative risk (RR) 2.5, 95% confidence interval (95% CI) 1.2 to 5.1) whereas exposure to chloroform was not. Exposure to BDCM of 20 microg/l and over was associated with decreased risks of cardiovascular anomalies (RR 0.3, 95% CI 0.2 to 0.7). There was a suggestion of an increased risk of chromosomal abnormalities associated with exposure to chloroform, and no evidence of any association between either trihalomethane compound and cleft defects. CONCLUSIONS: In this cohort, differences were found in the RR associated with exposure to chloroform and BDCM for each of the congenital anomalies under study. These findings point to the importance of examining specific byproduct compounds relative to risk for these birth outcomes and in particular implicate BDCM and other correlated disinfection byproducts in the aetiology of neural tube defects. PMID- 11404449 TI - Use of routinely collected data on trihalomethane in drinking water for epidemiological purposes. AB - OBJECTIVES: To explore the use of routinely collected trihalomethane (THM) measurements for epidemiological studies. Recently there has been interest in the relation between byproducts of disinfection of public drinking water and certain adverse reproductive outcomes, including stillbirth, congenital malformations, and low birth weight. METHOD: Five years of THM readings (1992--6), collected for compliance with statutory limits, were analysed. One water company in the north west of England, divided into 288 water zones, provided 15,984 observations for statistical analysis. On average each zone was sampled 11.1 times a year. Five year, annual, monthly, and seasonal variation in THMs were examined as well as the variability within and between zones. RESULTS: Between 1992 and 1996 the total THM (TTHM) annual zone means were less than half the statutory concentration, at approximately 46 microg/l. Differences in annual water zone means were within 7%. Over the study period, the maximum water zone mean fell from 142.2 to 88.1 microg/l. Mean annual concentrations for individual THMs (microg/l) were 36.6, 8.0, and 2.8 for chloroform, bromodichloromethane (BDCM), and dibromochloromethane (DBCM) respectively. Bromoform data were not analysed, because a high proportion of the data were below the detection limit. The correlation between chloroform and TTHM was 0.98, between BDCM and TTHM 0.62, and between DBCM and TTHM -0.09. Between zone variation was larger than within zone variation for chloroform and BDCM, but not for DBCM. There was only little seasonal variation (<3%). Monthly variation was found although there were no consistent trends within years. CONCLUSION: In an area where the TTHM concentrations were less than half the statutory limit (48 microg/l) chloroform formed a high proportion of TTHM. The results of the correlation analysis suggest that TTHM concentrations provided a good indication of chloroform concentrations, a reasonable indication of BDCM concentrations, but no indication of DBCM. Zone means were similar over the years, but the maximum concentrations reduced considerably, which suggests that successful improvements in treatment have been made to reduce high TTHM concentrations in the area. For chloroform and BDCM, the main THMs, the component between water zones was greater than variation within water zones and explained most of the overall exposure variation. Variation between months and seasons was low and showed no clear trends within years. The results indicate that routinely collected data can be used to obtain exposure estimates for epidemiological studies at a small area level. PMID- 11404450 TI - Effects on the nervous system in different groups of workers exposed to aluminium. AB - OBJECTIVE: To investigate possible neurotoxic effects in groups of aluminium pot room and foundry workers, aluminium welders, and a small group of workers exposed to aluminium in the production of flake powder. METHODS: Exposure to aluminium was evaluated with aluminium concentrations in blood and urine as well as a questionnaire. The groups exposed to aluminium were compared with a group of mild steel welders. Neurotoxic effects were studied with mood and symptom questionnaires and several psychological and neurophysiological tests. RESULTS: The pot room and foundry workers showed very low aluminium uptake as their aluminium concentrations in blood and urine were close to normal, and no effects on the nervous system were detected. The group of workers exposed to flake powder had high concentrations of aluminium in blood and urine, even higher than those of the aluminium welders. However, aluminium could not be shown to affect the functioning of the nervous system in flake powder producers. Although significant effects could not be shown in the present analysis of the data on welders, the performance of the welders exposed to high concentrations of aluminium was affected according to the analyses in the original paper from this group. CONCLUSIONS: For the pot room and foundry workers no effects related to the exposure to aluminium could be found. For the group of flake powder producers exposed for a short term no effects on the nervous systems were evident despite high levels of exposure. Due to the high concentrations of aluminium in the biological samples of this group, measures to reduce the exposure to aluminium are recommended, as effects on the central nervous system might develop after protracted exposures. However, this assumption needs to be verified in further studies. PMID- 11404451 TI - Mortality from cardiovascular diseases and exposure to inorganic mercury. AB - OBJECTIVE: To study the mortality from cardiovascular and other chronic non neoplastic diseases after long term exposure to inorganic mercury. Limited information is available on the effect of chronic exposure to mercury on the cardiovascular system. METHODS: The mortality was studied among 6784 male and 265 female workers from four mercury mines and mills in Spain, Slovenia, Italy, and the Ukraine. Workers were employed between 1900 and 1990; the follow up period lasted from the 1950s to the 1990s. The mortality of the workers was compared with national reference rates. RESULTS: Among men, there was a slight increase in overall mortality (standardised mortality ratio (SMR) 1.08, 95% confidence interval (95% CI) 1.04 to 1.12). An increased mortality was found from hypertension (SMR 1.46, 95% CI 1.08 to 1.93), heart diseases other than ischaemic (SMR 1.36, 95% CI 1.20 to 1.53), pneumoconiosis (SMR 27.1, 95% CI 23.1 to 31.6), and nephritis and nephrosis (SMR 1.55, 95% CI 1.13 to 2.06). The increase in mortality from cardiovascular diseases was not consistent among countries. Mortality from hypertension and other heart diseases increased with estimated cumulative exposure to mercury; mortality from ischaemic heart disease and cerebrovascular diseases increased with duration of employment, but not with estimated exposure to mercury. Results among women were hampered by few deaths. CONCLUSION: Despite limited quantitative data on exposure, possible confounding, and likely misclassification of disease, the study suggests a possible association between employment in mercury mining and refining and risk in some groups of cardiovascular diseases. PMID- 11404452 TI - Radiological progression and its predictive risk factors in silicosis. AB - OBJECTIVES: To investigate the risk factors predicting radiological progression in silicosis in a prospective cohort study of patients with silicosis who were previously exposed to silica from granite dust. METHODS: From among a total of 260 patients with silicosis contracted from granite work, 141 with available serial chest x ray films of acceptable quality taken over a period of 2 to 17 (mean 7.5) years, were selected for study. Ninety four (66.7%) had ended exposure 5 or more years previously (mean 10.1 years, maximum 28 years). Radiological progression was assessed by paired comparison of the initial and most recent radiographs, with two or more steps of increase in profusion of small opacities according to the 12 point scale of the International Labour Organisation (ILO) classification of radiographs of pneumoconiosis, taken from the majority reading by a panel of three independent readers. RESULTS: Overall, 37% of patients with silicosis had radiological evidence of progression. From the initial radiographs, 24 (31.6%) of those with radiological profusion category 1, 15 (37.5%) of those with radiological profusion category 2, and 13 (52%) of those with complicated silicosis (including all seven with category 3 profusion of small opacities) showed radiological progression. As expected, progression was more likely to be found after longer periods of follow up (the interval between the two chest x ray films) with a 20% increased odds of progression for every additional year of follow up. After adjustment for varying intervals of follow up, the probability of radiological progression was found to be significant if large opacities were present in the initial chest x ray film. Progression was also less likely to be found among those who had ended exposure to silica longer ago, although the result was of borderline significance (p=0.07). Tuberculosis was also associated with increased likelihood of progression (borderline significance). CONCLUSIONS: There is a high probability of radiological progression in silicosis after high levels of exposure to granite dust among workers who were followed up for up to 17 years. A significant risk factor is the extent of radiological opacities in the initial chest x ray film. The probability of progression is also likely to be reduced with longer periods after the end of exposure. PMID- 11404453 TI - Quantitative thermal perception thresholds relative to exposure to vibration. AB - OBJECTIVES: To assess the risk of disturbed thermal perception relative to exposure to vibration, to investigate a possible exposure-response relation and to analyse a possible relation between thermal perception and sensory symptoms. METHODS: The study was based on a cross section of 123 male workers exposed to vibration and 62 male workers who were not exposed. Thermal perception of cold, warmth, and heat pain was bilaterally determined from the thenar eminence by the method of limits. Perception of cold and warmth were also tested in the second digit. Personal energy equivalent exposure to vibration was measured for all subjects. Vibration was measured in accordance with International Standards Organisation (ISO) 5349 and assessed separately for the left and right hand. RESULTS: Combining exposure times and intensities gave the left hand an 0.80 exposure to vibration compared with the right. The risk of having contracted reduced thermal perception was increased at all test sites. The risk was higher for the thenar measurements than the finger measurements. A yearly extra contribution of 4000 mh/s(2) in cumulative exposure increases the risk of contracting a wider neutral zone by 18% (95% confidence interval (95% CI) 1.06 to 1.32) for the right and 18% (1.05 to 1.32) for the left hand side. Subjects with symptoms of nocturnal paraesthesia had a rate ratio (95% CI) of 2.80 (1.17 to 6.67) for the right hand and 2.72 (1.12 to 6.63) for the left hand for increased neutral zones at the thenar eminence. CONCLUSIONS: The results indicate thermal sensory impairment related to cumulative exposure to vibration. The effect appeared at vibration levels below the current guiding standard. Quantitative sensory testing of thermal perception offers the chance to assess this specific hazard to the peripheral sensorineural system associated with hand intensive work entailing vibration. PMID- 11404454 TI - What is "powder free"? Characterisation of powder aerosol produced during simulated use of powdered and powder free latex gloves. AB - OBJECTIVES: To characterise the distribution of particle size and mass of glove powder aerosol released from powdered and powder free non-sterile latex gloves under controlled conditions. METHODS: Gravimetric sampling and aerodynamic particle size analysis were performed during simulated use of gloves on a prosthetic hand in a chamber designed to minimise background particle concentrations. RESULTS: Aerosol was detectable for both powdered and powder free gloves under both aggressive and non-aggressive handling conditions. Most of the particles detected had aerodynamic diameter less than 10 microm. CONCLUSION: Powder free gloves were not entirely free of powder aerosol. Particles from both powdered and powder free gloves are sufficiently fine to penetrate into the thoracic region of the respiratory tract. PMID- 11404455 TI - Cancer after nuclear incidents. PMID- 11404456 TI - Exploring the DNA-binding specificities of zinc fingers with DNA microarrays. AB - A key step in the regulation of networks that control gene expression is the sequence-specific binding of transcription factors to their DNA recognition sites. A more complete understanding of these DNA-protein interactions will permit a more comprehensive and quantitative mapping of the regulatory pathways within cells, as well as a deeper understanding of the potential functions of individual genes regulated by newly identified DNA-binding sites. Here we describe a DNA microarray-based method to characterize sequence-specific DNA recognition by zinc-finger proteins. A phage display library, prepared by randomizing critical amino acid residues in the second of three fingers of the mouse Zif268 domain, provided a rich source of zinc-finger proteins with variant DNA-binding specificities. Microarrays containing all possible 3-bp binding sites for the variable zinc fingers permitted the quantitation of the binding site preferences of the entire library, pools of zinc fingers corresponding to different rounds of selection from this library, as well as individual Zif268 variants that were isolated from the library by using specific DNA sequences. The results demonstrate the feasibility of using DNA microarrays for genome-wide identification of putative transcription factor-binding sites. PMID- 11404457 TI - Identification and characterization of a second melanin-concentrating hormone receptor, MCH-2R. AB - Melanin-concentrating hormone (MCH) is a 19-aa cyclic neuropeptide originally isolated from chum salmon pituitaries. Besides its effects on the aggregation of melanophores in fish several lines of evidence suggest that in mammals MCH functions as a regulator of energy homeostasis. Recently, several groups reported the identification of an orphan G protein-coupled receptor as a receptor for MCH (MCH-1R). We hereby report the identification of a second human MCH receptor termed MCH-2R, which shares about 38% amino acid identity with MCH-1R. MCH-2R displayed high-affinity MCH binding, resulting in inositol phosphate turnover and release of intracellular calcium in mammalian cells. In contrast to MCH-1R, MCH 2R signaling is not sensitive to pertussis toxin and MCH-2R cannot reduce forskolin-stimulated cAMP production, suggesting an exclusive G(alpha)q coupling of the MCH-2R in cell-based systems. Northern blot and in situ hybridization analysis of human and monkey tissue shows that expression of MCH-2R mRNA is restricted to several regions of the brain, including the arcuate nucleus and the ventral medial hypothalamus, areas implicated in regulation of body weight. In addition, the human MCH-2R gene was mapped to the long arm of chromosome 6 at band 6q16.2-16.3, a region reported to be associated with cytogenetic abnormalities of obese patients. The characterization of a second mammalian G protein-coupled receptor for MCH potentially indicates that the control of energy homeostasis in mammals by the MCH neuropeptide system may be more complex than initially anticipated. PMID- 11404458 TI - Simple neural networks for the amplification and utilization of small changes in neuron firing rates. AB - I describe physiologically plausible "voter-coincidence" neural networks such that secondary "coincidence" neurons fire on the simultaneous receipt of sufficiently large sets of input pulses from primary sets of neurons. The networks operate such that the firing rate of the secondary, output neurons increases (or decreases) sharply when the mean firing rate of primary neurons increases (or decreases) to a much smaller degree. In certain sensory systems, signals that are generally smaller than the noise levels of individual primary detectors, are manifest in very small increases in the firing rates of sets of afferent neurons. For such systems, this kind of network can act to generate relatively large changes in the firing rate of secondary "coincidence" neurons. These differential amplification systems can be cascaded to generate sharp, "yes no" spike signals that can direct behavioral responses. PMID- 11404459 TI - Genetic and environmental factors modify bovine spongiform encephalopathy incubation period in mice. AB - The incubation period (IP) and the neuropathology of transmissible spongiform encephalopathies (TSEs) have been extensively used to distinguish prion isolates (or strains) inoculated into panels of inbred mouse strains. Such studies have shown that the bovine spongiform encephalopathy (BSE) agent is indistinguishable from the agent causing variant Creutzfeldt-Jakob disease (vCJD), but differs from isolates of sporadic CJD, reinforcing the idea that the vCJD epidemic in Britain results from consumption of contaminated beef products. We present a mouse model for genetic and environmental factors that modify the incubation period of BSE cross-species transmission. We have used two mouse strains that carry the same prion protein (PrP) allele, but display a 100-day difference in their mean IP following intracerebral inoculation with primary BSE isolate. We report genetic effects on IP that map to four chromosomal regions, and in addition we find significant factors of host environment, namely the age of the host's mother, the age of the host at infection, and an X-cytoplasm interaction in the host. PMID- 11404460 TI - Akt suppresses androgen-induced apoptosis by phosphorylating and inhibiting androgen receptor. AB - Whereas several apoptosis-related proteins have been linked to the antiapoptotic effects of Akt serine-threonine kinase, the search continues to explain the Akt signaling role in promoting cell survival via antiapoptotic effects. Here, we demonstrate that Akt phosphorylates the androgen receptor (AR) at Ser-210 and Ser 790. A mutation at AR Ser-210 results in the reversal of Akt-mediated suppression of AR transactivation. Activation of the phosphatidylinositol-3-OH kinase/Akt pathway results in the suppression of AR target genes, such as p21, and the decrease of androgen/AR-mediated apoptosis, which may involve the inhibition of interaction between AR and AR coregulators. Together, these findings provide a molecular basis for cross-talk between two signaling pathways at the level of Akt and AR-AR coregulators that may help us to better understand the roles of Akt in the androgen/AR-mediated apoptosis. PMID- 11404461 TI - Afipia felis induces uptake by macrophages directly into a nonendocytic compartment. AB - Afipia felis is a Gram-negative bacterium that causes some cases of human Cat Scratch Disease. A. felis can survive and multiply in several mammalian cell types, including macrophages, but the precise intracellular compartmentalization of A. felis-containing phagosomes is unknown. Here, we demonstrate that, in murine macrophages, most A. felis-containing phagosomes exclude lysosomal tracer loaded into macrophage lysosomes before, as well as endocytic tracer loaded after, establishment of an infection. Established Afipia-containing phagosomes possess neither early endosomal marker proteins [early endosome antigen 1 (EEA1), Rab5, transferrin receptor, trytophane aspartate containing coat protein (TACO)] nor late endosomal or lysosomal proteins [cathepsin D, beta-glucuronidase, vacuolar proton-pumping ATPase, rab7, mannose-6-phosphate receptor, vesicle associated membrane protein 8, lysosome-associated membrane proteins LAMP-1 and LAMP-2]. Those bacteria that will be found in a nonendosomal compartment enter the macrophage via an EEA1-negative compartment, which remains negative for LAMP 1. The smaller subpopulation of afipiae whose phagosomes will be part of the endocytic system enters into an EEA1-positive compartment, which also subsequently acquires LAMP-1. Killing of Afipia or opsonization with immune antibodies leads to a strong increase in the percentage of A. felis-containing phagosomes that interact with the endocytic system. We conclude that most phagosomes containing A. felis are disconnected from the endosome-lysosome continuum, that their unusual compartmentalization is decided at uptake, and that this compartmentalization requires bacterial viability. PMID- 11404462 TI - Copper-catalyzed oxidation of the recombinant SHa(29-231) prion protein. AB - Metal-catalyzed oxidation may result in structural damage to proteins and has been implicated in aging and disease, including neurological disorders such as Alzheimer's disease and amyotrophic lateral sclerosis. The selective modification of specific amino acid residues with high metal ion affinity leads to subtle structural changes that are not easy to detect but may have dramatic consequences on physical and functional properties of the oxidized protein molecules. PrP contains a histidine-rich octarepeat domain that binds copper. Because copper binding histidine residues are particularly prone to metal-catalyzed oxidation, we investigated the effect of this reaction on the recombinant prion protein SHaPrP(29-231). Using Cu2+/ascorbate, we oxidized SHaPrP(29-231) in vitro. Oxidation was demonstrated by liquid chromatography/mass spectrometry, which showed the appearance of protein species of higher mass, including increases in multiples of 16, characteristic of oxygen incorporation. Digestion studies using Lys C indicate that the 29-101 region, which includes the histidine-containing octarepeats, is particularly affected by oxidation. Oxidation was time- and copper concentration-dependent and was evident with copper concentrations as low as 1 microM. Concomitant with oxidation, SHaPrP(29-231) suffered aggregation and precipitation, which was nearly complete after 15 min, when the prion protein was incubated at 37 degrees C with a 6-fold molar excess of Cu2+. These findings indicate that PrP, a copper-binding protein, may be particularly susceptible to metal-catalyzed oxidation and that oxidation triggers an extensive structural transition leading to aggregation. PMID- 11404463 TI - A quantitative model for membrane fusion based on low-energy intermediates. AB - The energetics of a fusion pathway is considered, starting from the contact site where two apposed membranes each locally protrude (as "nipples") toward each other. The equilibrium distance between the tips of the two nipples is determined by a balance of physical forces: repulsion caused by hydration and attraction generated by fusion proteins. The energy to create the initial stalk, caused by bending of cis monolayer leaflets, is much less when the stalk forms between nipples rather than parallel flat membranes. The stalk cannot, however, expand by bending deformations alone, because this would necessitate the creation of a hydrophobic void of prohibitively high energy. But small movements of the lipids out of the plane of their monolayers allow transformation of the stalk into a modified stalk. This intermediate, not previously considered, is a low-energy structure that can reconfigure into a fusion pore via an additional intermediate, the prepore. The lipids of this latter structure are oriented as in a fusion pore, but the bilayer is locally compressed. All membrane rearrangements occur in a discrete local region without creation of an extended hemifusion diaphragm. Importantly, all steps of the proposed pathway are energetically feasible. PMID- 11404464 TI - Excessive tumor-elaborated VEGF and its neutralization define a lethal paraneoplastic syndrome. AB - Vascular endothelial growth factor (VEGF) is a potent endothelial cell mitogen and key regulator of both physiologic and pathologic (e.g., tumor) angiogenesis. In the course of studies designed to assess the ability of constitutive VEGF to block tumor regression in an inducible RAS melanoma model, mice implanted with VEGF-expressing tumors sustained high morbidity and mortality that were out of proportion to the tumor burden. Documented elevated serum levels of VEGF were associated with a lethal hepatic syndrome characterized by massive sinusoidal dilation and endothelial cell proliferation and apoptosis. Systemic levels of VEGF correlated with the severity of liver pathology and overall clinical compromise. A striking reversal of VEGF-induced liver pathology and prolonged survival were achieved by surgical excision of VEGF-secreting tumor or by systemic administration of a potent VEGF antagonist (VEGF-TRAP(R1R2)), thus defining a paraneoplastic syndrome caused by excessive VEGF activity. Moreover, this VEGF-induced syndrome resembles peliosis hepatis, a rare human condition that is encountered in the setting of advanced malignancies, high-dose androgen therapy, and Bartonella henselae infection. Thus, our findings in the mouse have suggested an etiologic role for VEGF in this disease and may lead to diagnostic and therapeutic options for this debilitating condition in humans. PMID- 11404465 TI - L-arginine-dependent suppression of apoptosis in Trypanosoma cruzi: contribution of the nitric oxide and polyamine pathways. AB - Until recently, a capacity for apoptosis and synthesis of nitric oxide *NO) were viewed as exclusive to multicellular organisms. The existence of these processes in unicellular parasites was recently described, with their biological significance remaining to be elucidated. We have evaluated L-arginine metabolism in Trypanosoma cruzi in the context of human serum-induced apoptotic death. Apoptosis was evidenced by the induction of DNA fragmentation and the inhibition of [3H]thymidine incorporation, which were inhibited by the caspase inhibitor Ac Asp-Glu-Val-aspartic acid aldehyde (DEVD-CHO). In T. cruzi exposed to death stimuli, supplementation with L-arginine inhibited DNA fragmentation, restored [3H]thymidine incorporation, and augmented parasite *NO production. These effects were inhibited by the *NO synthase inhibitor N(omega)-nitroarginine methyl ester (L-NAME). Exogenous *NO limited DNA fragmentation but did not restore proliferation rates. Because L-arginine is also a substrate for arginine decarboxylase (ADC), and its product agmatine is a precursor for polyamine synthesis, we evaluated the contribution of polyamines to limiting apoptosis. Addition of agmatine, putrescine, and the polyamines spermine and spermidine to T. cruzi sustained parasite proliferation and inhibited DNA fragmentation. Also, the ADC inhibitor difluoromethylarginine inhibited L-arginine-dependent restoration of parasite replication rates, while the protection from DNA fragmentation persisted. In aggregate, these results indicate that T. cruzi epimastigotes can undergo programmed cell death that can be inhibited by L arginine by means of (i) a *NO synthase-dependent *NO production that suppresses apoptosis and (ii) an ADC-dependent production of polyamines that support parasite proliferation. PMID- 11404466 TI - Synaptic sprouting increases the uptake capacities of motoneurons in amyotrophic lateral sclerosis mice. AB - Using adenoviruses encoding reporter genes as retrograde tracers, we assessed the capacity of motoneurons to take up and retrogradely transport adenoviral particles injected into the muscles of transgenic mice expressing the G93A human superoxide dismutase mutation, a model of amyotrophic lateral sclerosis. Surprisingly, transgene expression in the motoneurons was significantly higher in symptomatic mice than in control or presymptomatic mice. Using botulinum toxin to induce nerve sprouting at neuromuscular junctions, we showed that the unexpectedly high level of motoneurons retrograde transduction results, at least in part, from newly acquired uptake properties of the sprouts. These findings demonstrate the remarkable uptake properties of amyotrophic lateral sclerosis motoneurons in response to denervation and the rationale of using intramuscular injections of adenoviruses to overexpress therapeutic proteins in motor neuron diseases. PMID- 11404467 TI - A novel application of gene arrays: Escherichia coli array provides insight into the biology of the obligate endosymbiont of tsetse flies. AB - Symbiotic associations with microorganisms are pivotal in many insects. Yet, the functional roles of obligate symbionts have been difficult to study because it has not been possible to cultivate these organisms in vitro. The medically important tsetse fly (Diptera: Glossinidae) relies on its obligate endosymbiont, Wigglesworthia glossinidia, a member of the Enterobacteriaceae, closely related to Escherichia coli, for fertility and possibly nutrition. We show here that the intracellular Wigglesworthia has a reduced genome size smaller than 770 kb. In an attempt to understand the composition of its genome, we used the gene arrays developed for E. coli. We were able to identify 650 orthologous genes in Wigglesworthia corresponding to approximately 85% of its genome. The arrays were also applied for expression analysis using Wigglesworthia cDNA and 61 gene products were detected, presumably coding for some of its most abundant products. Overall, genes involved in cell processes, DNA replication, transcription, and translation were found largely retained in the small genome of Wigglesworthia. In addition, genes coding for transport proteins, chaperones, biosynthesis of cofactors, and some amino acids were found to comprise a significant portion, suggesting an important role for these proteins in its symbiotic life. Based on its expression profile, we predict that Wigglesworthia may be a facultative anaerobic organism that utilizes ammonia as its major source of nitrogen. We present an application of E. coli gene arrays to obtain broad genome information for a closely related organism in the absence of complete genome sequence data. PMID- 11404468 TI - Abortive base-excision repair of radiation-induced clustered DNA lesions in Escherichia coli. AB - It has been postulated that ionizing radiation produces a unique form of cellular DNA damage called "clustered damages" or "multiply damaged sites". Here, we show that clustered DNA damages are indeed formed in Escherichia coli by ionizing radiation and are converted to lethal double-strand breaks during attempted base excision repair. In wild-type cells possessing the oxidative DNA glycosylases that cleave DNA at repairable single damages, double-strand breaks are formed at radiation-induced clusters during postirradiation incubation and also in a dose dependent fashion. E. coli mutants lacking these enzymes do not form double strand breaks postirradiation and are substantially more radioresistant than wild type cells. Furthermore, overproduction of one of the oxidative DNA glycosylases in mutant cells confers a radiosensitive phenotype and an increase in the number of double-strand breaks. Thus, the effect of the oxidative DNA glycosylases in potentiating DNA damage must be considered when estimating radiation risk. PMID- 11404469 TI - Grafts of adenosine-releasing cells suppress seizures in kindling epilepsy. AB - Adenosine is an inhibitor of neuronal activity in the brain. The local release of adenosine from grafted cells was evaluated as an ex vivo gene therapy approach to suppress synchronous discharges and epileptic seizures. Fibroblasts were engineered to release adenosine by inactivating the adenosine-metabolizing enzymes adenosine kinase and adenosine deaminase. After encapsulation into semipermeable polymers, the cells were grafted into the brain ventricles of electrically kindled rats, a model of partial epilepsy. Grafted rats provided a nearly complete protection from behavioral seizures and a near-complete suppression of afterdischarges in electroencephalogram recordings, whereas the full tonic-clonic convulsions in control rats remained unaltered. Thus, the local release of adenosine resulting in adenosine concentrations <25 nM at the site of action is sufficient to suppress seizure activity and, therefore, provides a potential therapeutic principle for the treatment of drug-resistant partial epilepsies. PMID- 11404470 TI - Phenserine regulates translation of beta -amyloid precursor protein mRNA by a putative interleukin-1 responsive element, a target for drug development. AB - The reduction in levels of the potentially toxic amyloid-beta peptide (Abeta) has emerged as one of the most important therapeutic goals in Alzheimer's disease. Key targets for this goal are factors that affect the expression and processing of the Abeta precursor protein (betaAPP). Earlier reports from our laboratory have shown that a novel cholinesterase inhibitor, phenserine, reduces betaAPP levels in vivo. Herein, we studied the mechanism of phenserine's actions to define the regulatory elements in betaAPP processing. Phenserine treatment resulted in decreased secretion of soluble betaAPP and Abeta into the conditioned media of human neuroblastoma cells without cellular toxicity. The regulation of betaAPP protein expression by phenserine was posttranscriptional as it suppressed betaAPP protein expression without altering betaAPP mRNA levels. However, phenserine's action was neither mediated through classical receptor signaling pathways, involving extracellular signal-regulated kinase or phosphatidylinositol 3-kinase activation, nor was it associated with the anticholinesterase activity of the drug. Furthermore, phenserine reduced expression of a chloramphenicol acetyltransferase reporter fused to the 5'-mRNA leader sequence of betaAPP without altering expression of a control chloramphenicol acetyltransferase reporter. These studies suggest that phenserine reduces Abeta levels by regulating betaAPP translation via the recently described iron regulatory element in the 5'-untranslated region of betaAPP mRNA, which has been shown previously to be up-regulated in the presence of interleukin-1. This study identifies an approach for the regulation of betaAPP expression that can result in a substantial reduction in the level of Abeta. PMID- 11404471 TI - Crystal structure of an anticoagulant protein in complex with the Gla domain of factor X. AB - The gamma-carboxyglutamic acid (Gla) domain of blood coagulation factors is responsible for Ca2+-dependent phospholipid membrane binding. Factor X-binding protein (X-bp), an anticoagulant protein from snake venom, specifically binds to the Gla domain of factor X. The crystal structure of X-bp in complex with the Gla domain peptide of factor X at 2.3-A resolution showed that the anticoagulation is based on the fact that two patches of the Gla domain essential for membrane binding are buried in the complex formation. The Gla domain thus is expected to be a new target of anticoagulant drugs, and X-bp provides a basis for designing them. This structure also provides a membrane-bound model of factor X. PMID- 11404472 TI - In vivo activity in a catalytic antibody-prodrug system: Antibody catalyzed etoposide prodrug activation for selective chemotherapy. AB - Effective chemotherapy remains a key issue for successful cancer treatment in general and neuroblastoma in particular. Here we report a chemotherapeutic strategy based on catalytic antibody-mediated prodrug activation. To study this approach in an animal model of neuroblastoma, we have synthesized prodrugs of etoposide, a drug widely used to treat this cancer in humans. The prodrug incorporates a trigger portion designed to be released by sequential retro aldol/retro-Michael reactions catalyzed by aldolase antibody 38C2. This unique prodrug was greater than 10(2)-fold less toxic than etoposide itself in in vitro assays against the NXS2 neuroblastoma cell line. Drug activity was restored after activation by antibody 38C2. Proof of principle for local antibody-catalyzed prodrug activation in vivo was established in a syngeneic model of murine neuroblastoma. Mice with established 100-mm3 s.c. tumors who received one intratumoral injection of antibody 38C2 followed by systemic i.p. injections with the etoposide prodrug showed a 75% reduction in s.c. tumor growth. In contrast, injection of either antibody or prodrug alone had no antitumor effect. Systemic injections of etoposide at the maximum tolerated dose were significantly less effective than the intratumoral antibody 38C2 and systemic etoposide prodrug combination. Significantly, mice treated with the prodrug at 30-fold the maximum tolerated dose of etoposide showed no signs of prodrug toxicity, indicating that the prodrug is not activated by endogenous enzymes. These results suggest that this strategy may provide a new and potentially nonimmunogenic approach for targeted cancer chemotherapy. PMID- 11404473 TI - Ultrafast diffraction and structural dynamics: the nature of complex molecules far from equilibrium. AB - Studies of molecular structures at or near their equilibrium configurations have long provided information on their geometry in terms of bond distances and angles. Far-from-equilibrium structures are relatively unknown-especially for complex systems-and generally, neither their dynamics nor their average geometries can be extrapolated from equilibrium values. For such nonequilibrium structures, vibrational amplitudes and bond distances play a central role in phenomena such as energy redistribution and chemical reactivity. Ultrafast electron diffraction, which was developed to study transient molecular structures, provides a direct method for probing the nature of complex molecules far from equilibrium. Here we present our ultrafast electron diffraction observations of transient structures for two cyclic hydrocarbons. At high internal energies of approximately 4 eV, these molecules display markedly different behavior. For 1,3,5-cycloheptatriene, excitation results in the formation of hot ground-state structures with bond distances similar to those of the initial structure, but with nearly three times the average vibrational amplitude. Energy is redistributed within 5 ps, but with a negative temperature characterizing the nonequilibrium population. In contrast, the ring-opening reaction of 1,3-cyclohexadiene is shown to result in hot structures with a CC bond distance of over 1.7 A, which is 0.2 A away from any expected equilibrium value. Even up to 400 ps, energy remains trapped in large-amplitude motions comprised of torsion and asymmetric stretching. These studies promise a new direction for studying structural dynamics in nonequilibrium complex systems. PMID- 11404474 TI - Evidence for a circulating islet cell growth factor in insulin-resistant states. AB - Insulin resistance is a feature of many common disorders including obesity and type 2 diabetes mellitus. In these disorders, the beta-cells compensate for the insulin resistance for long periods of time with an increase in secretory capacity, an increase in beta-cell mass, or both. To determine whether the beta cell response might relate to a circulating growth factor, we have transplanted normal islets under the kidney capsule of normoglycemic insulin-resistant mice with two different models of insulin resistance: lean mice that have a double heterozygous deletion of the insulin receptor and insulin receptor substrate-1 (DH) or the obese, hyperglycemic ob/ob mice. In the grafts transplanted into both hosts, there was a marked increase in beta-cell mitotic activity and islet mass that was comparable with that observed in the endogenous pancreas. By contrast, islets of the DH mouse transplanted into normal mice showed reduced mitotic index. These data suggest the insulin resistance is associated with a circulating islet cell growth factor that is independent of glucose and obesity. PMID- 11404475 TI - Altered lymphocyte responses and cytokine production in mice deficient in the X linked lymphoproliferative disease gene SH2D1A/DSHP/SAP. AB - We have introduced a targeted mutation in SH2D1A/DSHP/SAP, the gene responsible for the human genetic disorder X-linked lymphoproliferative disease (XLP). SLAM associated protein (SAP)-deficient mice had normal lymphocyte development, but on challenge with infectious agents, recapitulated features of XLP. Infection of SAP mice with lymphocyte choriomeningitis virus (LCMV) or Toxoplasma gondii was associated with increased T cell activation and IFN-gamma production, as well as a reduction of Ig-secreting cells. Anti-CD3-stimulated splenocytes from uninfected SAP- mice produced increased IFN-gamma and decreased IL-4, findings supported by decreased serum IgE levels in vivo. The Th1 skewing of these animals suggests that cytokine misregulation may contribute to phenotypes associated with mutation of SH2D1A/SAP. PMID- 11404476 TI - Timing of metamorphosis and the onset of the negative feedback loop between the thyroid gland and the pituitary is controlled by type II iodothyronine deiodinase in Xenopus laevis. AB - Two important features of amphibian metamorphosis are the sequential response of tissues to different concentrations of thyroid hormone (TH) and the development of the negative feedback loop between the pituitary and the thyroid gland that regulates TH synthesis by the thyroid gland. At the climax of metamorphosis in Xenopus laevis (when the TH level is highest), the ratio of the circulating precursor thyroxine (T4) to the active form 3,5,3'-triiodothyronine (T3) in the blood is many times higher than it is in tissues. This difference is because of the conversion of T4 to T3 in target cells of the tadpole catalyzed by the enzyme type II iodothyronine deiodinase (D2) and the local effect (cell autonomy) of this activity. Limb buds and tails express D2 early and late in metamorphosis, respectively, correlating with the time that these organs undergo TH-induced change. T(3) is required to complete metamorphosis because the peak concentration of T4 that is reached at metamorphic climax cannot induce the final morphological changes. At the climax of metamorphosis, D2 expression is activated specifically in the anterior pituitary cells that express the genes for thyroid-stimulating hormone but not in the cells that express proopiomelanocortin. Physiological concentrations of T3 but not T4 can suppress thyrotropin subunit beta gene expression. The timing and the remarkable specificity of D2 expression in the thyrotrophs of the anterior pituitary coupled with the requirement for locally synthesized T3 strongly support a role for D2 in the onset of the negative feedback loop at the climax of metamorphosis. PMID- 11404477 TI - Light regulation of type IV pilus-dependent motility by chemosensor-like elements in Synechocystis PCC6803. AB - To optimize photosynthesis, cyanobacteria move toward or away from a light source by a process known as phototaxis. Phototactic movement of the cyanobacterium Synechocystis PCC6803 is a surface-dependent phenomenon that requires type IV pili, cellular appendages implicated in twitching and social motility in a range of bacteria. To elucidate regulation of cyanobacterial motility, we generated transposon-tagged mutants with aberrant phototaxis; mutants were either nonmotile or exhibited an "inverted motility response" (negative phototaxis) relative to wild-type cells. Several mutants contained transposons in genes similar to those involved in bacterial chemotaxis. Synechocystis PCC6803 has three loci with chemotaxis-like genes, of which two, Tax1 and Tax3, are involved in phototaxis. Transposons interrupting the Tax1 locus yielded mutants that exhibited an inverted motility response, suggesting that this locus is involved in controlling positive phototaxis. However, a strain null for taxAY1 was nonmotile and hyperpiliated. Interestingly, whereas the C-terminal region of the TaxD1 polypeptide is similar to the signaling domain of enteric methyl-accepting chemoreceptor proteins, the N terminus has two domains resembling chromophore binding domains of phytochrome, a photoreceptor in plants. Hence, TaxD1 may play a role in perceiving the light stimulus. Mutants in the Tax3 locus are nonmotile and do not make type IV pili. These findings establish links between chemotaxis like regulatory elements and type IV pilus-mediated phototaxis. PMID- 11404478 TI - Oxygen deprivation causes suspended animation in the zebrafish embryo. AB - Continuous exposure to oxygen is essential for nearly all vertebrates. We found that embryos of the zebrafish Danio rerio can survive for 24 h in the absence of oxygen (anoxia, 0% O2). In anoxia, zebrafish entered a state of suspended animation where all microscopically observable movement ceased, including cell division, developmental progression, and motility. Animals that had developed a heartbeat before anoxic exposure showed no evidence of a heartbeat until return to terrestrial atmosphere (normoxia, 20.8% O2). In analyzing cell-cycle changes of rapidly dividing blastomeres exposed to anoxia, we found that no cells arrested in mitosis. This is in sharp contrast to similarly staged normoxic embryos that consistently contain more than 15% of cells in mitosis. Flow cytometry analysis revealed that blastomeres arrested during the S and G2 phases of the cell cycle. This work indicates that survival of oxygen deprivation in vertebrates involves the reduction of diverse processes, such as cardiac function and cell-cycle progression, thus allowing energy supply to be matched by energy demands. PMID- 11404479 TI - The Friend of GATA proteins U-shaped, FOG-1, and FOG-2 function as negative regulators of blood, heart, and eye development in Drosophila. AB - Friend of GATA (FOG) proteins regulate GATA factor-activated gene transcription. During vertebrate hematopoiesis, FOG and GATA proteins cooperate to promote erythrocyte and megakaryocyte differentiation. The Drosophila FOG homologue U shaped (Ush) is expressed similarly in the blood cell anlage during embryogenesis. During hematopoiesis, the acute myeloid leukemia 1 homologue Lozenge and Glial cells missing are required for the production of crystal cells and plasmatocytes, respectively. However, additional factors have been predicted to control crystal cell proliferation. In this report, we show that Ush is expressed in hemocyte precursors and plasmatocytes throughout embryogenesis and larval development, and the GATA factor Serpent is essential for Ush embryonic expression. Furthermore, loss of ush function results in an overproduction of crystal cells, whereas forced expression of Ush reduces this cell population. Murine FOG-1 and FOG-2 also can repress crystal cell production, but a mutant version of FOG-2 lacking a conserved motif that binds the corepressor C-terminal binding protein fails to affect the cell lineage. The GATA factor Pannier (Pnr) is required for eye and heart development in Drosophila. When Ush, FOG-1, FOG-2, or mutant FOG-2 is coexpressed with Pnr during these developmental processes, severe eye and heart phenotypes result, consistent with a conserved negative regulation of Pnr function. These results indicate that the fly and mouse FOG proteins function similarly in three distinct cellular contexts in Drosophila, but may use different mechanisms to regulate genetic events in blood vs. cardial or eye cell lineages. PMID- 11404480 TI - Evolutionary EST analysis identifies rapidly evolving male reproductive proteins in Drosophila. AB - Sequence comparisons of genomes or expressed sequence tags (ESTs) from related organisms provide insight into functional conservation and diversification. We compare the sequences of ESTs from the male accessory gland of Drosophila simulans to their orthologs in its close relative Drosophila melanogaster, and demonstrate rapid divergence of many of these reproductive genes. Nineteen ( approximately 11%) of 176 independent genes identified in the EST screen contain protein-coding regions with an excess of nonsynonymous over synonymous changes, suggesting that their divergence has been accelerated by positive Darwinian selection. Genes that encode putative accessory gland-specific seminal fluid proteins had a significantly elevated level of nonsynonymous substitution relative to nonaccessory gland-specific genes. With the 57 new accessory gland genes reported here, we predict that approximately 90% of the male accessory gland genes have been identified. The evolutionary EST approach applied here to identify putative targets of adaptive evolution is readily applicable to other tissues and organisms. PMID- 11404481 TI - Stat3-mediated Myc expression is required for Src transformation and PDGF-induced mitogenesis. AB - Signal transducer and activator of transcription (STAT) proteins perform key roles in mediating signaling by cytokines and growth factors, including platelet derived growth factor (PDGF). In addition, Src family kinases activate STAT signaling and are required for PDGF-induced mitogenesis in normal cells. One STAT family member, Stat3, has been shown to have an essential role in cell transformation by the Src oncoprotein. However, the mechanisms by which STAT signaling pathways contribute to mitogenesis and transformation are not fully defined. We show here that disruption of Stat3 signaling by using dominant negative Stat3beta protein in NIH 3T3 fibroblasts suppresses c-Myc expression concomitant with inhibition of v-Src-induced transformation. Ectopic expression of c-Myc is able to partially reverse this inhibition, suggesting that c-Myc is a downstream effector of Stat3 signaling in v-Src transformation. Furthermore, c myc gene knockout fibroblasts are refractory to transformation by v-Src, consistent with a requirement for c-Myc protein in v-Src transformation. In normal NIH 3T3 cells, disruption of Stat3 signaling with dominant-negative Stat3beta protein inhibits PDGF-induced mitogenesis in a manner that is reversed by ectopic c-Myc expression. Moreover, inhibition of Src family kinases with the pharmacologic agent, SU6656, blocks Stat3 activation by PDGF. These findings, combined together, delineate the signaling pathway, PDGF --> Src --> Stat3 --> Myc, that is important in normal PDGF-induced mitogenesis and subverted in Src transformation. PMID- 11404482 TI - A pentacyclic reaction intermediate of riboflavin synthase. AB - The S41A mutant of riboflavin synthase from Escherichia coli catalyzes the formation of riboflavin from 6,7-dimethyl-8-ribityllumazine at a very low rate. Quenching of presteady-state reaction mixtures with trifluoroacetic acid afforded a compound with an absorption maximum at 412 nm (pH 1.0) that can be converted to a mixture of riboflavin and 6,7-dimethyl-8-ribityllumazine by treatment with wild type riboflavin synthase. The compound was shown to qualify as a kinetically competent intermediate of the riboflavin synthase-catalyzed reaction. Multinuclear NMR spectroscopy, using various 13C- and 15N-labeled samples, revealed a pentacyclic structure arising by dimerization of 6,7-dimethyl-8 ribityllumazine. Enzyme-catalyzed fragmentation of this compound under formation of riboflavin can occur easily by a sequence of two elimination reactions. PMID- 11404483 TI - Protein kinase Cdc15 activates the Dbf2-Mob1 kinase complex. AB - Exit from mitosis in budding yeast requires inactivation of cyclin-dependent kinases through mechanisms triggered by the protein phosphatase Cdc14. Cdc14 activity, in turn, is regulated by a group of proteins, the mitotic exit network (MEN), which includes Lte1, Tem1, Cdc5, Cdc15, Dbf2/Dbf20, and Mob1. The direct biochemical interactions between the components of the MEN remain largely unresolved. Here, we investigate the mechanisms that underlie activation of the protein kinase Dbf2. Dbf2 kinase activity depended on Tem1, Cdc15, and Mob1 in vivo. In vitro, recombinant protein kinase Cdc15 activated recombinant Dbf2, but only when Dbf2 was bound to Mob1. Conserved phosphorylation sites Ser-374 and Thr 544 (present in the human, Caenorhabditis elegans, and Drosophila melanogaster relatives of Dbf2) were required for DBF2 function in vivo, and activation of Dbf2-Mob1 by Cdc15 in vitro. Although Cdc15 phosphorylated Dbf2, Dbf2-Mob1, and Dbf2(S374A/T544A)-Mob1, the pattern of phosphate incorporation into Dbf2 was substantially altered by either the S374A T544A mutations or omission of Mob1. Thus, Cdc15 promotes the exit from mitosis by directly switching on the kinase activity of Dbf2. We propose that Mob1 promotes this activation process by enabling Cdc15 to phosphorylate the critical Ser-374 and Thr-544 phosphoacceptor sites of Dbf2. PMID- 11404484 TI - Positive selection of an MHC class-I restricted TCR in the absence of classical MHC class I molecules. AB - The H-2Ld alloreactive 2C T cell receptor (TCR) is commonly considered as being positively selected on the H-2Kb molecule. Surprisingly, 2C TCR+ CD8+ single positive T cells emerge in massive numbers in fetal thymic organ culture originating from 2C transgenic, H-2KbD(b-/-) (2C+KbD(b-/-)) but not in fetal thymic organ culture from beta2-microglobulin(-/-) 2C transgenic animals. Mature CD8+ T cells are observed in newborn but not in adult 2C+KbD(b-/-) mice. These CD8+ T cells express the alpha4beta7 integrin, which allows them to populate the intestine, a pattern of migration visualized by intrathymic injection of FITC and subsequent accrual of FITC-labeled lymphocytes in the gut. We conclude that the 2C TCR is reactive not only with H-2Ld and H-2Kb, but also with nonclassical MHC class I products to enable positive selection of 2C+ T cells in the fetal and newborn thymus and to support their maintenance in the intestine. PMID- 11404488 TI - Are DNA ploidy and epidermal growth factor receptor prognostic factors for untreated ovarian cancer? A prospective study. AB - To identify prognostic factors for untreated ovarian cancer, DNA ploidy, proliferative index (P.I.) and epidermal growth factor receptor (EGFR) expression were analyzed in a prospective series of 40 patients with ovarian cancer and 7 patients with borderline malignant ovarian tumor followed up for 5 years or more (median, 77 months). The frequency of aneuploid cells was 53.8% (21/39) in ovarian cancer and 14.3% (1/7) in borderline malignancy. There was no significant association between DNA ploidy and the clinicopathologic findings, in which aneuploid ovarian cancer was more common among advanced tumors. The S-phase fraction and P.I. value were higher in the patients with aneuploid tumors (p = 0.076). EGFR expression was detected in 76.9% (30/39) of ovarian cancers and 42.9% (3/7) of borderline malignant ovarian tumors, and the mean EGFR level was 5.8 +/- 12.1 (range: 0-49.5) and 28.3 +/- 71.1 (range: 0-189.4) fmol/mg protein, respectively. There was no correlation between EGFR expression and DNA ploidy, P.I., and clinicopathologic findings analyzed. The 5-year survival rate in patients with aneuploid tumors was significantly worse in patients with ovarian cancer (p = 0.0165, log-rank test). No significant relationship was shown between P.I., EGFR expression, and 5-year survival. Cox multivariate analysis showed that DNA ploidy, P.I., and EGFR expression are not associated with the risk of death (p = 0.5917, p = 0.9924, and p = 0.6840, respectively), although clinical stage shows a significant relationship (p = 0.0027). Our data showed that DNA ploidy is significantly related to the prognosis by univariate analysis, but DNA ploidy, P.I., and EGFR expression were not independent prognostic factors for the untreated ovarian cancer. PMID- 11404489 TI - Angiogenesis and apoptosis-related protein (p53, bcl-2, and bax) expression versus response of gastric adenocarcinomas to paclitaxel and carboplatin chemotherapy. AB - The role of angiogenesis and apoptosis-related proteins in defining response to chemotherapy is poorly understood. We examined the microvessel density (MVD) and the expression of p53, bcl-2, and bax proteins in a series of 28 locally advanced gastric adenocarcinomas, treated with paclitaxel and carboplatin. A strong cytoplasmic reactivity in more than 10% of cancer cells was recorded in 25% of cases for p53 protein, and in 14% and 64% of cases for bcl-2 and bax proteins, respectively. Microvessel density was assigned in three categories: low (<35), medium (35-60), and high (>60). Tumors of medium MVD showed a significantly higher response rate compared with those of high or low MVD (p = 0.01 and 0.001, respectively), and prognosis was significantly better in this group of patients with medium MVD tumors (p < 0.02). Loss of bax protein expression was somewhat more frequent in tumors resistant to chemotherapy, but this difference was not of statistical significance. Nuclear p53 reactivity was associated with higher MVD (p = 0.02). The expression of p53 and bcl-2 did not influence the outcome of treatment. The present study suggests that although apoptosis-related proteins may have a role in defining response to taxanes, parameters related to tumors' vasculature, such as drug availability or angiogenic tissue regeneration, may be equally important. PMID- 11404490 TI - Sequential chemoradiotherapy with docetaxel, cisplatin, and 5-fluorouracil in patients with locally advanced head and neck cancer. AB - The purpose of this phase II trial was to evaluate the toxicity of a sequential chemoradiotherapy approach using docetaxel, cisplatin, and 5-fluorouracil (5-FU) (DCF) with granulocyte colony-stimulating factor support in previously untreated patients with locally advanced head and neck cancer (HNC). Secondary endpoints included preliminary assessment of response. Patients with locally advanced HNC, a World Health Organization performance status 0 to 2, and no prior history of chemotherapy or radiotherapy were included. Treatment consisted of docetaxel 80 mg/m2 (1-hour infusion) on day 1, cisplatin 40 mg/m2 (1-hour infusion) on days 2 and 3, and 5-fluorouracil 1,000 mg/m2 (24-hour continuous infusion), on days 1 to 3, repeated every 28 days for a maximum of 4 cycles per patient. All patients received granulocyte colony stimulating factors subcutaneously between days 4 and 9. Radiation therapy (RT) to the primary tumor site and neck lymph nodes was planned within 5 weeks of the last cycle of chemotherapy. The primary tumor site received 60 to 70 Gy. Twenty patients (median age 56 years, range: 40-72 years) received a total of 60 cycles of DCF. The median number of cycles was 3 (range: 1 4 cycles). All patients were evaluable for toxicity and response. The most common acute nonhematologic toxicities from DCF induction chemotherapy included alopecia, mucositis, peripheral sensory neuropathy, onycholysis, and asthenia. Febrile neutropenia developed in two patients and grade IV diarrhea in one patient. There were no treatment-related deaths. The overall response rate (RR) after DCF induction chemotherapy was 90% (95% confidence interval [CI]: 76.8 103.1%). After the completion of RT, the overall RR was 95% with a complete response rate of 73% (95% CI: 49.9-90.1%). Organ preservation was achieved in eight patients with laryngeal cancer and one patient with base of tongue involvement. After a median follow-up of 36 months (range: 5-43 months) the median disease-free and overall survival have not been reached yet. The 1- and 2 year survival rates were 85% and 60%, respectively. Sequential chemoradiotherapy with DCF and growth factor support is feasible and very active, with durable responses in patients with locally advanced head and neck cancer. Further evaluation of this modality is justified in the context of a clinical trial. PMID- 11404491 TI - Phase II study of epirubicin, mitomycin C, and 5-fluorouracil in hormone refractory prostatic carcinoma. AB - Management of metastatic prostatic carcinoma when it becomes refractory to hormonal therapy is controversial, and no standard treatment exists. Nevertheless, chemotherapy for hormone-refractory prostatic carcinoma (HRPC) has shown some advantages compared with the best supportive care. In a prospective phase II study, we evaluated the combination of epirubicin (E), mitomycin C (MMC), and 5-fluorouracil (5-FU) in patients with HRPC. Twenty-eight patients with HRPC were treated with a combination of E (30 mg/m2), 5-FU (750 mg/m2), and MMC (5 mg/m2) day 1 and 2, every 4 weeks. Treatment was continued until evidence of disease progression or excessive toxicity. Patients were monitored with serial measurements of prostate-specific antigen (PSA). Forty-seven percent of the patients exhibited a reduction of serum PSA concentration and an objective response; 38% exhibited disease stability, and 15% had disease progression. Toxicity was substantial. The median time to progression was 7.3 months (range, 1.7-16.8 months) and median survival was 14.5 months (range, 1.6-38.4 months). Performance status improved in 80% of patients, and bone pain was relieved in 70%. Thus the combination of E, MMC, and 5-FU shows activity in the treatment of HRPC, giving substantial palliation of symptoms. In one patient, PSA values remained low even when the tumor had progressed. PMID- 11404492 TI - Primary cutaneous large B-cell lymphoma of the legs: a distinct clinical pathologic entity treated with CD20 monoclonal antibody (rituximab). AB - Primary cutaneous large B-cell lymphoma of the legs (PCLBLL) is most commonly diagnosed in the elderly, and is generally confined to the lower parts of one or sometimes both legs. Despite treatment with radiotherapy, relapses and extracutaneous involvement can occur, and unlike other low-grade cutaneous-B-cell non-Hodgkin's lymphomas (NHLs), the prognosis is variable, with an estimated 5 year survival rate of 58%. This report describes the case of an 81-year-old man who was diagnosed with PCLBLL. Staging evaluation did not reveal NHL elsewhere. The patient declined recommendations to receive cytotoxic chemotherapy. Instead, he was treated with anti-CD20 monoclonal therapy (rituximab) and his cutaneous lesions completely regressed during a 16-week period. This report suggests that rituximab is a therapeutic option for those patients with PCLBLL who may not be good candidates to receive radiation therapy or chemotherapy. Long-term follow-up and greater experience with rituximab in a variety of clinical settings will ultimately determine the appropriate role of this costly, but relatively safe, antibody-based therapy for CD20+ expressing NHLs. PMID- 11404493 TI - Super high-dose intraarterial cisplatin infusion under percutaneous pelvic perfusion with extracorporeal chemofiltration for advanced uterine cervical carcinoma: I. Analysis for pharmacokinetics, tumor response, and toxicity of platinum. AB - The present study was designed to elucidate the clinical feasibility of a new intraarterial infusion system with an extracorporeal charcoal chemofiltration circuit, which is expected to achieve a super high-dose cisplatin pelvic perfusion with a limited systemic exposure to platinum. After inferior vena cava isolation was percutaneously achieved by balloon catheter technique, cisplatin (140-240 mg/m2) was administered by selective intrauterine arterial infusion, with inferior and superior gluteal arterial embolization. The platinum-containing blood was pumped through an extracorporeal charcoal chemofiltration circuit. Pharmacokinetics, tumor response, and toxicity of platinum under this system were studied in 14 patients with locally advanced uterine cervical carcinoma. Extracorporeal charcoal filters significantly (p < 0.05) reduced the prefilter area under concentration-time curve of plasma-free platinum by 86.7 +/- 5.2% at postfilter site and 76.3 +/- 6.6% at peripheral circulation, respectively. Although all adverse effects were mild under this system, tumor response and tissue platinum concentrations were augmented dose dependently with the administration of cisplatin. The extracorporeal chemofiltration system achieved a super high-dose cisplatin pelvic perfusion with the minimal adverse effects, allowing further cisplatin dose escalation with further augmented tumor response. This will contribute to the reduction in the extent of disease of locally advanced uterine cervical carcinoma. PMID- 11404494 TI - Super high-dose intraarterial cisplatin infusion under percutaneous pelvic perfusion with extracorporeal chemofiltration for advanced uterine cervical carcinoma: II. Its impact on clinical response and subsequent surgery. AB - The present pilot study was conducted to investigate the clinical efficacy of super high-dose intraarterial cisplatin infusion with percutaneous pelvic perfusion under extracorporeal chemofiltration (PPPEC) for locally advanced uterine cervical carcinoma. Cisplatin (140-240 mg/m2) was infused in uterine arteries in a neoadjuvant setting in 20 patients under the PPPEC system twice during a 2-week interval. Fourteen of 17 patients in whom reduction of the disease (tumor downstaging) was confirmed underwent radical surgery. Despite the tumor downstaging, the remaining three patients had poor PS and the other three showed insufficient stage regression. Clinical responses, histologic responses, and surgical review were studied. The rate of overall tumor response (complete response plus partial response), tumor downstaging, overall histologic response, and radical surgery performance after the second course of PPPEC were 95.0%, 85.0%, 95.0%, and 70.0%, respectively. Curative surgery, defined as negative carcinoma cells in surgical margins, was achieved in 85.7% of the cases, whereas the rate of complete surgery defined as negative carcinoma cells both in surgical margins and regional lymph nodes was 42.9%. With 42 months of median follow-up time, 3 of the 14 surgical patients died of the original disease, and the remaining 9 patients are in recurrence-free survival, whereas 2 patients are alive with disease. PPPEC achieved a high frequency of rapid tumor downstaging of locally advanced uterine cervical carcinoma without severe adverse effects and resulted in the favorable performance of the subsequent radical surgery and prognosis. PMID- 11404495 TI - Phase I-II study of irinotecan in combination with mitomycin C in patients with advanced gastrointestinal cancer. AB - This phase I-II study was conducted to determine the maximum tolerated dose and optimal schedule of a combination of irinotecan (CPT 11) and mitomycin C (MMC) in a population of previously treated patients with gastrointestinal malignancies. Four cohorts of patients were recruited with MMC given at 8 mg/m2 for the first 3 levels together with irinotecan at 300 mg/m2, 325 mg/m2, and 350 mg/m2; the fourth dose level was given with MMC at 10 mg/m2 and irinotecan at 325 mg/m2. All treatment was repeated at 21-day intervals. The dose-limiting toxicity was hematologic (thrombocytopenia at level 4), and the recommended doses for subsequent phase II studies are MMC 8 mg/m2 with irinotecan 325 mg/m2. Evidence of efficacy was seen at all dose levels examined and justifies further exploration of this combination in a less heavily pretreated patient population. PMID- 11404496 TI - Linear accelerator based radiosurgery as a salvage treatment for skull base and intracranial invasion of recurrent nasopharyngeal carcinomas. AB - Nasopharyngeal carcinoma is a common disease entity in Taiwan. It can spread in the prestyloid, retrostyloid compartments, parapharyngeal space, and skull base, and induce paralysis of cranial nerves. We have treated more than 1,500 cases in the past 14 years. Since 1994, we have treated 11 cases of nasopharyngeal carcinomas with skull base invasion by linear accelerator-based stereotactic radiosurgery. There were six male and five female patients. The mean age was 48.4 years. Seven patients had symptoms of low cranial nerves. For these patients, conformal radiotherapy plus brachytherapy was used for the masses beyond the skull base. Stereotactic radiosurgery with a dose of 10 Gy to 19 Gy for one treatment was used for the masses invading the skull base. Eight patients died in follow-up periods from 5 months to 2 years 7 months. Approximately half of patients revealed good response of tumor to radiosurgery. Two patients demonstrated no response of tumors to radiosurgery. Stereotactic radiosurgery may be an adjuvant treatment for recurrent nasopharyngeal carcinomas with skull base and intracranial invasion. PMID- 11404497 TI - Neoadjuvant chemotherapy in inoperable, locally advanced, and inflammatory breast carcinoma: a pilot study of MTT assay in vitro and outcome analysis of 10 patients. AB - Patients with inoperable, locally advanced, and inflammatory breast carcinoma (LAIBC), whether with supraclavicular lymph nodes (SLN) or not (stage IIIB and IV), usually carry an overall poor prognosis. The current treatment for these patients is by means of combined modality, including preoperative chemotherapy. This strategy has led to a substantial improvement in clinical response, making some patients operable, and even making breast conservative surgery possible. However, the long-term results still are not promising. The aim of this pilot study was to determine the efficacy of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay in vitro in directing chemotherapy (including preoperative adjuvant chemotherapy and postoperative adjuvant chemotherapy) for these patients. Between June 1994 and March 1997, 10 patients with inoperable LAIBC, whether with SLN or not, were enrolled. During the period of the combined therapy modalities, the neoadjuvant chemotherapy was adopted for three cycles according to the results of chemosensitivity in vitro by MTT assay. Then a modified radical or radical mastectomy was performed, which was followed by radiotherapy and further postoperative adjuvant chemotherapy with the same regimen as that of neoadjuvant chemotherapy. All patients had been followed up from the beginning of neoadjuvant chemotherapy to the end of October 1999. Two patients had clinical complete response (CRs), with one having pathologic CR in both breast tumor and axillary lymph node, and the other having pathologic CR in axillary lymph node. The other eight patients had partial response. By the time of analysis, six patients had been dead of relapse or progression. Among the four patients who were still alive, one had local relapse, one had distant metastatic disease, and the other two had no evident disease. By retrieving from MEDLINE before 1999, the authors learned that this is the first pilot study of neoadjuvant chemotherapy for inoperable LAIBC using MTT assay to predict the chemosensitivity in vitro. Compared with conventional chemotherapy, the clinical response and long-term results seem to be more encouraging. PMID- 11404498 TI - Phase II trial of the use of paclitaxel and gemcitabine as a salvage treatment in metastatic breast cancer. AB - The purpose of this study was to evaluate gemcitabine plus paclitaxel in heavily pretreated patients with metastatic breast cancer (MBC). Patients with MBC with second or third relapse to anthracycline-containing regimens received a 3-hour infusion of paclitaxel 175 mg/m2 on day 1, and gemcitabine 1.0 g/m2 on days 1, 8, and 15, every 28 days. Because of unacceptable thrombocytopenia seen in the first 5 patients, the gemcitabine schedule was changed to days 1 and 8 (G-1,8) for the remainder of the study, every 21 days. Twenty-nine patients (median age, 46 years; range, 32-68 years) received 137 cycles (median: 4 per patient). The regimen was well tolerated. World Health Organization grades III and IV thrombocytopenia were observed in 5 (18.5%) of the first 27 cycles (G-1,8,15), and in 6 (5.4%) of the 110 subsequent cycles (G-1,8)--p = 0.04 for the difference between schedules. Five patients had grade I and two had grade III neuropathy. Eight patients had grade III neutropenia, two had grade IV neutropenia associated with fever (G-1,8,15), and eight had grades I and II myalgia and fatigue. There were 16 (55%) objective responses (95% CI 36-73%); 5 (17%) complete responses, 11 (38%) partial responses (95% CI 3-30% and 19-56%, respectively), and 6 (20.5%) patients with stable disease. Median response duration was 8 months (range, 4-26 months). Median overall survival was 12 months (range, 4-28+ months), and 1-year and 2-year survival rates were 45% and 30%, respectively. This phase II study demonstrated a manageable toxicity profile with the gemcitabine day 1, 8 schedule in combination with paclitaxel and significant and promising activity in heavily pretreated patients with MBC. A confirmatory phase III trial is warranted. PMID- 11404499 TI - Management of neurocytomas: case report and review of the literature. AB - The purpose of this report is to review the available literature on the presentation, pathology, and treatment of central nervous system (CNS) neurocytomas. A case report of an extraventricular neurocytoma is presented along with a comprehensive literature search of patients with a diagnosis of CNS neurocytoma. CNS neurocytomas are rare neoplasms, with fewer than 240 cases reported in the literature. The majority of neurocytomas are found in the ventricular system of the brain. Immunohistochemistry is frequently used to help distinguish this tumor from other CNS neoplasms. MIB-1 proliferation index is commonly used in an attempt to predict biologic behavior. Little is known about the management of patients with this tumor, because most reports are from the pathologic literature and contain sparse information regarding clinical management. Neurocytomas are rare CNS tumors with varied biologic behavior. MIB-1 index may help direct adjuvant therapy. An excellent prognosis can be expected if a gross total resection is achieved. Postoperative radiation therapy (RT) may be considered after subtotal resection. Otherwise, RT is an option for medically inoperable or recurrent disease. PMID- 11404500 TI - Temporary reversal by topotecan of marked insulin resistance in a patient with myelodysplastic syndrome: case report and possible mechanism for tumor necrosis factor alpha (TNF-alpha)-induced insulin resistance. AB - Tumor necrosis factor-alpha (TNF-alpha) is an important mediator of insulin resistance in obesity and diabetes through its ability to decrease the tyrosine kinase activity of the insulin receptor. We report here a remarkable degree of insulin resistance in a patient with adult respiratory distress syndrome and myelodysplasia. PMID- 11404501 TI - Phase II trial of toremifene in androgen-independent prostate cancer: a Penn cancer clinical trials group trial. AB - Toremifene has antiestrogenic and estrogenic properties in vitro and in vivo. In addition, it may have antiangiogenesis and antimicrotubule properties at higher doses. Studies have demonstrated the efficacy of this agent in the treatment of metastatic breast cancer. We performed a phase II trial of toremifene in patients with androgen-independent prostate cancer (AIPC). Patients with an increasing prostate-specific antigen level despite castrate testosterone levels and antiandrogen withdrawal were eligible. Patients could not have received prior salvage hormonal therapy or chemotherapy. Patients received toremifene at 300 mg/m2/d orally (maximum dose 640 mg/d). Fifteen patients were treated. Patients received treatment for a median of 13 weeks (range, 4-30 weeks). The median age was 72 years (range, 58-80 years). The median Eastern Cooperative Oncology Group performance status was 0. The treatment was well tolerated and toxicity was mild. Two patients had grade III hepatic toxicity; one had grade III hyperglycemia. There were no treatment-related deaths. No objective responses were demonstrated. In summary, toremifene is not effective therapy for AIPC at the dose and schedule evaluated in this trial. PMID- 11404502 TI - Multivariate prognostic analysis of stage I(E) primary non-Hodgkin's lymphomas of the nasal cavity. AB - From January 1968 to December 1997, a total of 71 patients with stage I(E) (Ann Arbor staging system, 1971) primary non-Hodgkin's lymphomas of the nasal cavity received treatment in the Cancer Center of Sun Yat-Sen University of Medical Sciences. Thirty-seven lesions were limited to the nasal cavity (limited I(E)), whereas the other 34 were extended to the structure out of the nasal cavity (extended I(E)). Forty-four patients were treated with radiochemotherapy and 27 with radiotherapy alone. Kaplan-Meier methods were used in the survival analysis. Multivariate analysis was carried out using the Cox proportional hazard model. The 5- and 10-year survival rates were 71.85% and 59.67% for the patients with a complete response to irradiation, and both were 13.89% for the patients with residue lesions (p = 0.0004). The 5- and 10-year survival rates were 69.81% and 56.72% for limited I(E), and 40.65% and 35.57% for extended I(E) (p = 0.0047). The prognosis was better for those younger than 44 years (p = 0.0003). The 10 year survival rates for radiotherapy alone and combined radiochemotherapy are 52% and 75% for limited I(E) versus 37.58% and 45% for extended I(E) (p = 0.0644). B symptoms did not significantly affect clinical outcome (p = 0.729). Multivariate analysis showed that complete response of local lesion after radiotherapy, invasion of the primary tumor to adjacent structures, and patients' age were independent prognostic factors. Our study showed that radiotherapy is the main treatment method for the primary non-Hodgkin's lymphomas of the nasal cavity; the addition of chemotherapy may improve long-term survival. The local tumor response to radiotherapy, whether the extranasal structures were invaded, and patients' age were independent prognostic factors. PMID- 11404503 TI - Twice-daily prophylactic cranial irradiation for patients with limited disease small-cell lung cancer with complete response to chemotherapy and consolidative radiotherapy: report of a single institutional phase II trial. AB - Prophylactic cranial irradiation (PCI) has been demonstrated to significantly reduce the incidence of brain relapse from limited disease small-cell lung cancer (LD SCLC), but concerns about neurologic toxicity remain. The purpose of this report was to update a phase II institutional trial that explored the impact of twice-daily PCI on neurologic toxicity as well as outcome for this group of patients. All eligible subjects had documented complete response to induction chemotherapy and consolidative chest irradiation. The whole brain was treated with twice-daily fractions of 1.5 Gy with megavoltage irradiation to an approximate total dose of 30.0-36.0 Gy. Although not devised as a randomized study, approximately half of the eligible patients declined the protocol enrollment of their own volition and were retrospectively evaluated as a "historical" control group regarding the incidence of brain metastases. Fifteen patients accepted twice-daily PCI, with 12 deferring treatment. Median follow-up was 20 months. Disease-free survival at 2 years was 54% with twice-daily PCI versus 0% without any PCI (p = 0.013). Overall survival at 2 years was 62% with twice-daily PCI versus 23% without PCI (p = 0.032). No statistically significant neurologic deterioration was detected in the PCI group posttreatment. Thus, twice daily PCI should be considered for patients with LD SCLC who achieve a complete response to chemoirradiation. A multi-institutional randomized trial would be necessary before making definitive recommendations. PMID- 11404504 TI - Gemcitabine in advanced pancreatic cancer: a phase II trial. AB - The 5-year survival for pancreatic cancer is usually less than 5%, and no treatment has demonstrated consistent effect on patient survival and disease related symptoms. Early studies with gemcitabine suggested a modest antitumor activity with significant improvement in disease-related symptoms. This phase II study reports the activity of gemcitabine on 33 consecutive patients with unresectable pancreatic carcinoma. Twenty-three patients had metastatic and 10 locally advanced unresectable disease. Twenty-six patients had not received any previous treatment and seven had received first-line chemotherapy with 5 fluorouracil. Gemcitabine 1,000 mg/m2 was administered intravenously in 30 minutes in the first cycle once weekly for up to 7 weeks followed by 1 week rest; then in subsequent cycles, once weekly for 3 of every 4-week cycle. Four patients obtained partial response (12%). Fifteen patients (45%) had stable disease with a median duration of 32 weeks (range: 16-75 weeks), and 14 patients experienced progressive disease. Median duration of response was 34.5 weeks (range: 19-50 weeks). Median survival was 33 weeks (range: 2-91 weeks). All 4 responding patients and 14 of 15 (93%) patients with stable disease had improvement in performance status and decrease in daily analgesic requirement. Toxicity was mild and mainly consisted of moderate and rapidly reversible myelosuppression. We conclude that gemcitabine chemotherapy was very well tolerated and determined a significant clinical improvement with modest antitumoral activity in patients with advanced pancreatic cancer. PMID- 11404505 TI - Primary thyroid lymphoma: a retrospective analysis of prognostic factors and treatment outcome for localized intermediate and high grade lymphoma. AB - Non-Hodgkin's lymphoma presenting in the thyroid gland is uncommon. A review of the King Faisal Specialist Hospital and Research Centre (KFSH & RC) experience was performed to assess treatment outcome and prognostic factors in this rare extranodal presentation of localized lymphoma. Sixty patients treated at KFSH & RC between 1975 and 1995 were identified, and their records were reviewed retrospectively. Eight patients who had stage III or IV disease, low grade, or did not complete their prescribed treatment were excluded from the study. There were 38 female and 14 male patients with a median age of 59.5 years at the time of diagnosis (range: 10-87 years). Thirty-five of the 52 patients underwent diagnostic partial or total thyroidectomy at other institutions based on a preoperative assumption of thyroid carcinoma. All 52 patients had non-Hodgkin's lymphoma of intermediate (94%) or high (6%) grade. Detailed staging was carried out in all patients; 16 patients (31%) had disease confined to the thyroid gland (stage IE), whereas 36 (69%) had associated disease in cervical lymph nodes and/or the mediastinum (stage IIE) disease. All patients were treated with curative intent. A total of 18 patients (35%) were treated with a single-modality treatment--radiotherapy alone in 2, chemotherapy alone in 13, and surgery alone in the remaining 3 patients. The majority of patients (34/52; 65%) were treated with a combined-modality approach. The overall relapse-free survival (RFS) and overall survival (OS) at 5 years were 72% and 88%, respectively. There were no significant differences in outcome between those treated with single-modality and those with combined-modality therapy. A univariate analysis showed that the presence of mediastinal lymph node involvement was the most important prognostic factor affecting both RFS and OS. Patients with Hashimoto thyroiditis and without "B" symptoms were found to have a significantly higher RFS without influence on the OS. However, patients who had a good performance status (PS) of 0, 1, and 2 were found to have a significantly higher overall survival in comparison to those with poor performance status. Age, sex, stage, histology, lactic acid dehydrogenase level, tumor bulk, and the treatment modality were not found to correlate with RFS or OS. Mediastinal involvement and PS were found to be the most important independent prognostic factors influencing RFS and OS. PMID- 11404506 TI - Solitary splenic metastasis: case report and review of the literature. AB - Solitary splenic lesion is a rare presentation of a previous or current malignancy. We describe a case of an isolated splenic lesion caused by endometrial carcinoma and summarize all the published reports on solitary splenic metastasis. PMID- 11404507 TI - Synchronous isolated splenic metastasis from colon carcinoma. AB - We report the first case known to us of a synchronous isolated splenic metastasis from colon carcinoma in a 52-year-old woman operated on splenectomy, left colectomy, and ileal resection. The patient died of diffuse carcinomatosis 1 year after the operation. Splenectomy for isolated splenic metastasis from colon carcinoma is justified, and serum tumor markers are useful to detect metastases early during the follow-up, as in our report. PMID- 11404508 TI - Palliative management of metastatic tumors involving the psoas muscle: case reports and review of the literature. AB - Seven case reports of patients with metastatic tumors in the psoas muscle are described. Reasons for the rarity of this neoplastic disease complication are considered. The efficacy of radiotherapy in providing palliation of symptomatic patients is examined. PMID- 11404509 TI - Spinal cord compression, infection, and unknown primary cancers. AB - The management of a patient with spinal cord compression can be complicated and challenging; however, this challenge becomes even more pronounced if the patient presents without a known cancer diagnosis in the context of progressive neurologic symptoms. Unless potential causes such as infection/tuberculosis are included in the differential diagnosis of an apparent unknown primary, then the correct diagnosis may be incorrectly determined. PMID- 11404510 TI - Docetaxel and liver dysfunction: is it absolutely contraindicated? PMID- 11404512 TI - Thomas Sedlacek, M.D., 1942-2000: a memorial. PMID- 11404511 TI - Primary breast lymphoma and bone marrow transplantation. PMID- 11404513 TI - Cytokine profile and immunomodulation in asymptomatic human T-lymphotropic virus type 1-infected blood donors. AB - The modulation of the immune response has been used as therapy for clinical disorders associated with human T-lymphotropic virus type 1 (HTLV-1) infection. In this study, the cytokine profile was evaluated in 26 asymptomatic HTLV-1 blood donors. Additionally, both the cell responsible for producing interferon-gamma (IFN-gamma) and the role of exogenous interleukin (IL)-10 in downregulating IFN gamma production were studied. Cytokine levels were determined in supernatants of unstimulated lymphocyte cultures by enzyme-linked immunosorbent assay. The levels of IFN-gamma, tumor necrosis factor-alpha, IL-5, and IL-10 were higher in supernatants of the lymphocyte cultures taken from HTLV-1-infected donors than in those taken from healthy subjects. Although depletion of CD8+ T cells and natural killer cells did not affect IFN-gamma production, depletion of CD4+ T cells significantly decreased IFN-gamma production. Furthermore, at a concentration of 2 ng/ml, IL-10 had only a minimum effect on IFN-gamma production, although at high concentrations (100 ng/ml), IL-10 decreased IFN-gamma production by 50% in HTLV-1-infected individuals. These data indicate that both T helper 1 and T helper 2 cytokines are elevated in HTLV-1 infection and that IL-10 in high concentrations modulates IFN-gamma production in these patients. PMID- 11404514 TI - A novel recombinant marker virus assay for comparing the relative fitness of hiv 1 reverse transcriptase variants. AB - A novel recombinant marker virus assay (RMVA) has been developed to perform growth competition assays for assessing fitness of HIV-1. This assay allowed the generation of replication-competent viruses by homologous recombination of polymerase chain reaction (PCR)-derived reverse transcriptase (RT) coding sequences in RT-deleted proviral clones of HIV-1 in which the nef gene was replaced by the Salmonella typhimurium histidinol dehydrogenase (hisD) or human heat-stable placental alkaline phosphatase (PLAP) gene (pHIVDeltaRTBalIDeltanefhisD and pHIVDeltaRTBalIDeltanefPLAP, respectively). The proportion of a given RT species in a mixed culture was determined by quantifying the linked hisD or PLAP marker gene using real-time PCR. The RMVA was tested by comparing the relative fitness of wild-type and lamivudine-resistant recombinant viruses. The RMVA reproducibly detected differences in the fitness of these two viruses in growth competition assays. With appropriate modification of the recombination vectors, the RMVA should be useful for analyzing the fitness of viruses resistant to protease, integrase, or fusion inhibitors and should be applicable in clinical research. PMID- 11404515 TI - Long-term impact of highly active antiretroviral therapy on HIV-related health care costs. AB - CONTEXT: Highly active antiretroviral therapy (HAART) is associated with decreased opportunistic infections, hospitalization, and HIV-related health care costs over relatively short periods of time. We have previously demonstrated that decreases in total HIV cost are proportional to penetration of protease inhibitor therapy in our clinic. OBJECTIVE: To determine the effects of HAART on HIV health care use and costs over 44 months. SETTING: A comprehensive HIV service within a Veterans Affairs Medical Center. DESIGN: A cost-effectiveness analysis of HAART. MAIN OUTCOME MEASUREMENTS: The mean monthly number of hospital days, infectious diseases clinic visits, emergency room visits, non-HIV-related outpatient visits, inpatient costs, and antiretroviral treatment costs per patient were determined by dividing these during the period from January 1995 through June 1998 into four intervals. Viral load tests were available from October 1996. Cost-effectiveness of HAART was evaluated by determining the costs of achieving an undetectable viral load over time. RESULTS: Mean monthly hospitalization and associated inpatient costs decreased and remained low 2 years after the introduction of protease inhibitors (37 hospital days per 100 patients). Total cost decreased from $1905 per patient per month during the first quarter to $1090 per patient per month in the third quarter but increased to $1391 per patient per month in the fourth quarter. Antiretroviral treatment costs increased throughout the entire observation period from $79 per patient per month to $518 per patient per month. Hospitalization costs decreased from $1275 per patient per month in the first quarter to less than $500 per patient per month in each of the third and fourth quarters. The percentage of patients with a viral load <500 copies/mL increased from 21% in October 1996 to 47% in June of 1997 (p =.014). The cost of achieving an undetectable viral load decreased from $4438 per patient per month to $2669 per patient per month, but this trend did not reach statistical significance (p =.18). CONCLUSIONS: After an initial decrease, there was an increase in the total monthly cost of caring for HIV patients. Cost increases were primarily due to antiretroviral treatment costs, but these costs were offset by a marked decrease in inpatient-related costs. Increases in costs were not related to antiretroviral treatment failures as measured by the proportion of patients with low or undetectable viral loads. The cost of achieving an undetectable viral load remained stable despite increases in the cost of procuring antiretroviral agents. PMID- 11404516 TI - Association of provider and patient characteristics with HIV-infected women's antiretroviral therapy regimen. AB - OBJECTIVE: We explored the effect of patient and provider factors on the type of antiretroviral regimen among women receiving therapy. PATIENTS: Five hundred ninety-five New York State nonpregnant HIV+ women with full Medicaid eligibility and at least 1 month of a prescribed antiretroviral regimen in federal fiscal years (FFY) 1997-1998 and intervals in FFY 1997-1998, who had delivered a liveborn baby within 5 years. MEASUREMENTS: From pharmacy claims in 4 6-month intervals in FFY 1997-1998, data were extracted on (1) an acceptable > or = 2 antiretroviral combination regimen per expert guidelines; and (2) a highly active regimen, including a protease inhibitor or nonnucleoside analog (highly active antiretroviral therapy [HAART]). RESULTS: Of 1514 woman-6-month intervals with filled antiretroviral prescriptions, 82% had an acceptable regimen, and of 1246 woman-6-month intervals on acceptable antiretroviral therapy, half demonstrated the use of HAART. Adjusted odds ratios (AORs) of acceptable antiretroviral therapy were higher (p < .05) for HIV specialty care (AOR = 1.71 for one or two visits; AOR = 2.10 for 3+ visits) or HIV clinical trials site care (AOR = 1.43; 95% confidence interval [CI]: 1.01, 2.04). Among women on acceptable antiretroviral regimens, those aged older than 25 years (AOR = 1.69; CI: 1.13, 2.53) or who were high school graduates (AOR = 1.50; CI: 1.09, 2.06) had higher odds of HAART. Methadone-treated women had twofold and nearly threefold higher AORs of acceptable antiretroviral regimens and HAART, respectively, than current drug users. CONCLUSION: Provider HIV expertise is associated with receipt of an acceptable antiretroviral regimen in women, although receipt of HAART is affected more by age, education, and current drug abuse. Methadone treatment seems to improve access to acceptable antiretroviral regimens as well as to HAART. PMID- 11404517 TI - Lipodystrophy in HIV-infected children is associated with high viral load and low CD4+ -lymphocyte count and CD4+ -lymphocyte percentage at baseline and use of protease inhibitors and stavudine. AB - Alterations in regional fat, often associated with abnormalities in lipid and insulin metabolism, have been reported in HIV-infected adults. To determine whether similar abnormalities occur in children with HIV, patterns of change in regional body fat distribution were determined by dual energy x-ray absorptiometry in 28 prepubertal HIV-infected children. Eight (29%) children experienced lipodystrophy (LD), defined as extremity lipoatrophy together with trunk fat accumulation. Despite a mean body weight increase of 2.9 +/- 2.4 kg, children with LD experienced a mean loss of total fat in contrast to children without LD who increased total fat (-0.151 +/- 0.324 versus 0.981 +/- 1.041 kg; p <.01). Children with LD had significantly higher levels of HIV RNA and lower CD4 count and percentage at baseline. LD was associated with use of protease inhibitors or stavudine, (odds ratio [OR], 7.0, 95% confidence interval [CI], 1.1 45.2, p =.04; OR, 9.0, 95% CI, 1.4-59.8, p =.03, respectively). This observational study suggests that during a time in childhood when accumulation of extremity and trunk fat is expected, some HIV-infected children experience changes in fat distribution that are similar to HIV-associated LD reported in adults. Studies to determine whether HIV-infected children with changes in regional fat also experience increases in "atherogenic" lipids and insulin resistance as described in adults with HIV-associated LD are warranted. PMID- 11404518 TI - Pulmonary infiltrates in HIV-infected patients in the highly active antiretroviral therapy era in Spain. AB - OBJECTIVE: To study the incidence, etiology, and outcome of pulmonary infiltrates (PIs) in HIV-infected patients and to evaluate the yield of diagnostic procedures. DESIGN: Prospective observational study of consecutive hospital admissions. SETTING: Tertiary hospital. PATIENTS: HIV-infected patients with new onset radiologic PIs from April 1998 to March 1999. METHODS: The study protocol included chest radiography, blood and sputum cultures, serologic testing for "atypical" causes of pneumonia, testing for Legionella urinary antigen, testing for cytomegalovirus antigenemia, and bronchoscopy in case of diffuse or progressive PIs. RESULTS: One hundred two episodes in 92 patients were recorded. The incidence of PIs was 18 episodes per 100 hospital admission-years (95% confidence interval [CI]: 15-21). An etiologic diagnosis was achieved in 62 cases (61%). Bacterial pneumonia (BP), Pneumocystis carinii pneumonia (PCP), and mycobacteriosis were the main diagnoses. The incidences of BP and mycobacteriosis were not statistically different in highly active antiretroviral therapy (HAART) versus non-HAART patients. The incidence of PCP was lower in those receiving HAART (p =.011), however. Nine patients died (10%). Independent factors associated with higher mortality were mechanical ventilation (odds ratio [OR] = 83; CI: 4.2-1,682), age >50 years (OR = 23; CI: 2-283), and not having an etiologic diagnosis (OR = 22; CI: 1.6-293). CONCLUSIONS: Pulmonary infiltrates are still a frequent cause of hospital admission in the HAART era, and BP is the main etiology. There was no difference in the rate of BP and mycobacteriosis in HAART and non-HAART patients. Not having an etiologic diagnosis is an independent factor associated with mortality. PMID- 11404519 TI - Relation between CD4 cell counts and HIV RNA levels at onset of opportunistic infections. AB - OBJECTIVE: To evaluate the relation between CD4 and HIV RNA levels at the onset of specific opportunistic infections (OIs) in HIV-infected patients. DESIGN AND METHODS: The OIs occurring between June 1996 and December 1998 were retrospectively reviewed, considering only the episodes of major and minor OIs in patients with simultaneously available CD4 and plasma HIV RNA determinations before clinical onset who had been untreated or on stable antiretroviral therapy (ART) for at least 2 months. RESULTS: Two hundred seventy-four episodes of different OIs were considered in 216 patients; the median CD4 count was 35 cells/mm3 (range: 0-1154 cells/mm(3)), and the median HIV RNA count was 5.1 log cp/mL (range: < 1.9-6.7 log copies/ml). The different HIV RNA levels were significantly associated with different OIs regardless of CD4 and ART (p < .0001), even when only those occurring in patients with a CD4 count of < or = 50 cells/mm(3) were considered (p = .0049). Kaposi sarcoma, esophageal candidiasis, oropharyngeal candidiasis, and Mycobacterium avium complex disease were associated with significantly above-average median HIV RNA levels, and varicella zoster virus infection was associated with below-average levels. CONCLUSIONS: Different OIs are associated at their onset with significantly different HIV RNA levels, regardless of CD4 cell counts and ART. PMID- 11404520 TI - Incidence and determinants of bacterial infections in HIV-positive patients receiving anti-Pneumocystis carinii/Toxoplasma gondii primary prophylaxis within a randomized clinical trial. AB - We assessed the incidence and determinants of bacteremia, pneumonia, and sinusitis/otitis in HIV-positive people receiving cotrimoxazole (CTX) or dapsone pyrimethamine (DP) for primary prophylaxis of Pneumocystis carinii pneumonia (PCP) and toxoplasmic encephalitis (TE) within a randomized clinical trial. In total, 244 patients were randomized: 122 were assigned to CTX and 122 to DP. In the cohort, 22 bacteremia, 63 pneumonia, and 39 sinusitis/otitis cases were observed. Incidence rates of bacteremia, pneumonia, and sinusitis/otitis as well as the 2-year probability of remaining free from any bacterial infection were not significantly different between the two groups. At multivariate analysis, the risks of developing bacteremia and pneumonia were found to be independently increased by the use of a central venous catheter (hazard ratio [HR], 4.48; p <.05 and HR, 4.13; p <.01, respectively) and by hospitalization (HR, 28.82; p <.05 and HR, 10.15; p <.05, respectively). In conclusion, CTX at the dosage employed for primary PCP/TE prophylaxis does not seem to protect against bacterial infections more than second-line DP. PMID- 11404521 TI - Plasma zinc, copper, copper:zinc ratio, and survival in a cohort of HIV-1 infected homosexual men. AB - A prospective cohort study of 121 HIV-1-positive homosexual men was conducted in Miami, Florida, U.S.A. to evaluate the associations between plasma zinc and copper levels and mortality. Plasma zinc and copper levels were measured at baseline and then at semiannual visits. Zinc inadequacy and copper inadequacy were defined as plasma zinc levels <75 (microg/dl) and plasma copper levels <85 (microg/dl), respectively. HIV-1-related deaths were confirmed by review of death certificates. Cox proportional hazards regression models with time-dependent covariates were used to estimate the relative risks of zinc and copper inadequacy on mortality. Over the average course of the 3.3-year follow-up, 19 participants (16%) died of HIV-1-related causes. After adjustment for potential confounders, including low CD4+ cell counts and antiretroviral therapy, zinc inadequacy and copper:zinc ratio >1 (i.e., plasma copper level greater than plasma zinc level) were associated with increased mortality (relative risks [RRs]; 95% confidence intervals [CIs], 4.98, 1.30-19.00 and 8.28, 1.03-66.58, respectively). A negative association was also observed between plasma zinc levels and mortality (RR 0.94; 95% CI, 0.91-0.98). Plasma levels of copper were not significantly associated with mortality. These results suggest that plasma zinc inadequacy or the plasma copper:zinc ratio may be useful predictors of survival in HIV-1 infection. The latter appears to be a stronger predictor. PMID- 11404522 TI - Assessment of the performance of a rapid, lateral flow assay for the detection of antibodies to HIV. AB - Rapid HIV assays have recently been shown to have important applications for various testing situations, including early identification of infected individuals, to allow intervention strategies in a clinically relevant time frame. A rapid, lateral flow, HIV-1/2/O assay was evaluated using 2,000 serum or plasma samples from various risk groups and geographic locations, including HIV-1 and HIV-2 positive sera from five countries. Two U.S. Food and Drug Administration (FDA)-licensed screening assays and a FDA-licensed confirmatory assay were used as reference tests. The rapid assay exhibited a near-perfect sensitivity (99.2%) and an excellent specificity (99.9%). Moreover, its analytical sensitivity was found to be better than most FDA-licensed enzyme linked immunosorbent assays (ELISAs), detecting infection at the same time as the most sensitive ELISA in two of five seroconversion panels, and at the same time or earlier than four of five ELISAs in all five panels. We conclude that this rapid assay is a suitable test for the detection of HIV infection that could be particularly useful in developing countries where facilities may not support the use of instrumentation. PMID- 11404523 TI - Injection-related risk behaviors in young urban and suburban injection drug users in Chicago (1997-1999). AB - We compared injection-related risk practices between urban and suburban injection drug users (IDUs) in a large cross-sectional sample of young IDUs. From 1997 to 1999, we recruited 700 active IDUs aged 18 to 30 years in Chicago and its suburbs. A suburban residence was reported by 38% of participants. Participants were interviewed at four urban locations and screened for HIV and hepatitis C virus antibodies. Receptive sharing of syringes and other paraphernalia by urban and suburban IDUs in the preceding 6 months was compared using univariable and multivariable models. Sharing injection paraphernalia in the total sample was high, with 50% of participants reporting receptive syringe sharing and 70% reporting sharing cotton, cookers, and/or rinse water. After adjusting for demographic characteristics, injection settings, frequency, and duration of injection as well as ease of acquiring new syringes, suburban IDUs were significantly more likely than urban IDUs to share syringes (adjusted odds ratio = 1.7; 95% confidence interval: 1.1-2.5); however, the likelihood of sharing cotton, cookers, or rinse water was roughly equal. Despite overall higher risk profiles among suburban IDUs, HIV and hepatitis C prevalence levels were significantly lower than among urban participants. Current high levels of injection risk behaviors in suburban groups represent a potential for rapid dissemination of infection. PMID- 11404524 TI - HIV-positive men's sexual practices in the context of self-disclosure of HIV status. AB - OBJECTIVE: To examine whether disclosure of HIV-positive status to sex partners at risk for HIV infection is associated with safer sex practices and to examine the prevalence and correlates of specific disclosure/sexual behavior patterns. METHODS: Cross-sectional assessment of 206 HIV-positive men (41% homosexual, 35% bisexual, 24% heterosexual) sampled randomly at an outpatient HIV clinic in Los Angeles, who reported that their most recent sex partner was HIV-negative or of unknown serostatus. Unsafe sex was defined as unprotected anal or vaginal intercourse with that partner. RESULTS: Twenty-five percent of the men engaged in unsafe sex, and 48% of the total sample withheld disclosure from the partner. The prevalence of safer sex was not significantly higher among disclosers than among nondisclosers (unadjusted odds ratio = 1.29; 95% confidence interval: 0.69-2.45), and disclosure was not significantly associated with safer sex in any of 25 demographic or partner subgroups examined in the study. In the full sample, 40% of the men disclosed and engaged in safer sex (informed protection), 35% withheld disclosure and engaged in safer sex (uninformed protection), 12% informed their partner and engaged in unsafe sexual behavior (informed exposure), and 13% withheld disclosure and engaged in unsafe sex (uninformed exposure). Risky behavior patterns were associated with using alcohol/drugs before sex, having an HIV-unknown partner, being less emotionally involved with one's partner, and testing seropositive in the previous 3 years. CONCLUSIONS: Interventions for seropositive men that focus primarily on increasing disclosure of serostatus to sex partners may not reduce the prevalence of unsafe sex. Interventions are needed to address the social and psychologic processes that give rise to risky behavior patterns in HIV-infected men. Improved substance abuse counseling also may be needed. PMID- 11404525 TI - Substance use among men who have sex with men: comparison with a national household survey. AB - OBJECTIVE: Compare substance use among men who have sex with men (MSM) at high risk for HIV infection to a nationally representative sample of heterosexual men. METHODS: Compare data from surveys of 3,212 MSM recruited for participation in a Vaccine Preparedness Study (VPS) with an age-standardized group of 2481 single, urban-dwelling men from the 1995 National Household Survey on Drug Abuse (NHSDA). RESULTS: Except for alcohol, relative risk (RR [95% confidence interval (CI)]) for use of any substance was higher in the VPS than the National Household Survey on Drug Abuse (NHSDA) (3.64 [3.01-4.42]). Drugs with the highest relative risks were "poppers" (21.6 [15.2-30.8]), sedatives (6.98 [2.46-19.8]), hallucinogens (6.14 [4.61-8.17]), tranquilizers (4.99 [2.96-8.42]), and stimulants (4.47 [3.58 5.58]). RR was higher for weekly use of poppers (33.5 [12.5-89.6]), stimulants (2.75 [1.79-4.22]), marijuana (2.37 [1.93-2.92]), and cocaine (2.24 [1.32-3.79]); and for daily use of marijuana (1.49 [1.08-2.05]). CONCLUSIONS: Participants in the VPS used more substances than a group of age-standardized, single, urban dwelling men from the NHSDA. In view of previous data showing that substance use can be associated with unprotected sex, assessing substance use among MSM at high risk for HIV infection is an important component of risk reduction efforts. PMID- 11404526 TI - Peptides derived from salivary thrombospondin-1 replicate its anti-HIV effect: potential role in microbicide development. PMID- 11404527 TI - Temporal changes of progression to AIDS in the era of highly active antiretroviral therapy: Lazio Region, Italy, 1988 to June 2000. PMID- 11404528 TI - Screening for HIV infection in indigent women with newly diagnosed cervical cancer. PMID- 11404529 TI - Oral mucosal lesions and HIV viral load in the Women's Interagency HIV Study (WIHS). PMID- 11404530 TI - Zidovudine genotypic resistance in HIV-1-infected newborns in the French perinatal cohort. AB - A retrospective study was set up to investigate the frequency of zidovudine (ZDV) resistant HIV-1 in infected newborns after ZDV prophylaxis in the French Perinatal Cohort study. Nucleotide sequence analysis was carried out from 34 infants' isolates and 18 maternal plasma samples. Mutations related to ZDV resistance were found in the HIV-1 reverse transcriptase in 7 of 34 children (20%). Evidence of mother-child transmission of ZDV-resistant HIV-1 was found in 4 cases. Phylogenetic analysis showed that 14 of 34 HIV-1 isolates from the infants belonged to non-B subtypes. The presence of ZDV resistance-encoding mutations in the newborn isolates was associated with a longer total duration of exposure to ZDV. In a context of a wide HIV-1 variability, ZDV resistance can be one of the factors contributing to mother-child transmission. PMID- 11404531 TI - CCR5 and CXCR4 expression on memory and naive T cells in HIV-1 infection and response to highly active antiretroviral therapy. AB - OBJECTIVE: To measure CCR5 and CXCR4 chemokine receptor expression on CD4 and CD8 T cells in HIV-1 infection and to relate levels to the distribution of CD45RO memory and CD45RA-naive subsets, measures of disease activity, and response to highly active antiretroviral therapy (HAART). DESIGN: Fourteen untreated HIV-1 infected patients, 18 patients at 3-to 4-weeks after beginning HAART, and 35 uninfected control subjects were studied. METHODS: Four-color cytofluorometry with appropriate conjugated monoclonal antibodies (mAbs) was performed to define CD45RA and CD45RO subsets of CD4 and CD8 T cells and measure their expression of CCR5, CXCR4, and CD38. RESULTS: HIV-1-infected patients had higher CCR5 levels and lower CXCR4 levels on CD4 and CD8 T cells and their CD45RO/CD45RA subsets than control subjects did. However, CCR5 elevation was statistically significant only for CD4 T cells and their subsets, and CXCR4 depression was significant for CD8 T cells and their subsets (and for CD4:CD45RO cells). The elevation of CCR5 and depression of CXCR4 were not due to shifts in CD45RO/CD45RA subset proportions but to upregulation or downregulation within the subsets. CCR5 elevation on CD4 T cells was significantly restored toward normal by HAART, but the CXCR4 depression was not. CCR5 expression but not CXCR4 expression correlated with other measures of immunodeficiency (CD4 T-cell levels), active infection (viral load), and cellular activation (CD38). CONCLUSIONS: CCR5 elevation is a concomitant of immune activation and viral replication that occurs in HIV-1 infection, but the relation of CXCR4 depression to severity of infection, disease progression, and response to therapy remains undefined. PMID- 11404532 TI - A randomized, open-label, comparative trial of zidovudine plus lamivudine versus zidovudine plus lamivudine plus didanosine in antiretroviral-naive HIV-1-infected Thai patients. AB - OBJECTIVE: To assess the efficacy and tolerability of a triple nucleoside reverse transcriptase inhibitor combination of zidovudine, lamivudine, and didanosine therapy. DESIGN: A randomized open-label trial. PATIENTS: Antiretroviral-naive HIV-infected patients with CD4+ cell counts of 100 to 500 cells/microl. METHODS: A total of 106 patients were randomly assigned to 300 mg of zidovudine (200 mg for body weight <60 kg) twice daily plus 150 mg of lamivudine twice daily plus 200 mg of didanosine (125 mg for body weight <60 kg) twice daily (n = 53) or to zidovudine plus lamivudine (n = 53) for 48 weeks. MAIN OUTCOME MEASURES: Degree and duration of reduction of HIV-1 RNA load and increase in CD4+ cell counts from baseline and development of drug-related toxicities. RESULTS: At 48 weeks, triple drug therapy showed greater declines in plasma HIV-RNA levels from the beginning of treatment than double drug therapy (1.86 vs. 1.15 log10 copies/ml, respectively; p <.001). The proportions of patients with HIV-RNA <50 copies/ml in an intention-to-treat analysis were 54.7% (29 of 53 patients) and 11.3% (6 of 53 patients) in the triple and double drug therapy, respectively (p =.001). There was no significant difference in increase of CD4 count. CONCLUSION: Triple drug therapy with zidovudine, lamivudine, and didanosine was significantly more effective in inducing sustained immunologic and virologic responses than the double combination of zidovudine and lamivudine. PMID- 11404533 TI - The effect of nevirapine in combination with nelfinavir in heavily pretreated HIV 1-infected patients: a prospective, open-label, controlled, randomized study. AB - The purpose of the current study was to determine the efficacy and safety of nevirapine combined with nelfinavir and two nucleoside reverse transcriptase inhibitors (NRTIs) in patients previously exposed to highly active antiretroviral therapy (HAART). In a prospective, open-label, randomized study, 56 HIV-infected adults who had received HAART, including saquinavir hard gel capsule, ritonavir, or indinavir, were randomly assigned to receive nevirapine in addition to nelfinavir and two NRTIs. The proportion of patients who achieved an undetectable viral load (plasma HIV-RNA <200 copies/ml) at weeks 24 and 36 was significantly higher in the nevirapine group than in the control group (55% and 52% vs. 22% and 22%; p =.015 and p =.047). No differences in CD4 cell count or clinical outcome were observed. In the nevirapine group, 17% of patients discontinued treatment because of rashes. We conclude that the addition of nevirapine, when switching from one protease inhibitor-containing regimen to one containing nelfinavir, has a substantial effect on viral suppression. PMID- 11404534 TI - Prospective evaluation of the effect of initiating indinavir-based therapy on insulin sensitivity and B-cell function in HIV-infected patients. AB - OBJECTIVE: To determine whether initiation of antiretroviral therapy that includes the protease inhibitor indinavir causes insulin resistance or abnormal B cell function in study subjects with HIV infection. METHODS: Nonwasted, HIV infected study subjects who did not have concurrent diabetes were prospectively evaluated by oral and intravenous glucose tolerance testing at baseline, at 2 weeks after starting indinavir monotherapy, and at another 6 weeks after initiating indinavir-based triple-therapy. RESULTS: Mean CD4 count at entry was 282 cells/microl and median HIV RNA was 33,000 copies/ml; all experienced a virologic response. Fasting glucose increased from 83.2 +/- 3.7 mg/dl at baseline to 86.8 +/- 3.2 at week 2 and 91.7 +/- 3.5 at week 8 (p =.003). Insulin sensitivity by minimal model analysis decreased by 30.5% over 8 weeks, from 3.83 +/- 0.63 min-1 per microU/ml x 10-4 to 3.09 +/- 0.53 at week 2 and 2.66 +/- 0.35 at week 8 (p =.01). Insulin secretion by the acute insulin response to intravenous glucose did not change (baseline 822 +/- 283 microU/ml x min, week 8 880 +/- 289; p = 0.4), and the insulin response to oral glucose (30 minute insulin:glucose ratio) fell from 1.69 +/- 0.54 microU/ml per mg/dl at baseline to 1.18 +/- 0.34 at week 8 (p =.05). CONCLUSION: During 8 weeks of indinavir-based therapy, fasting glucose increased and insulin sensitivity decreased, without a compensatory increase in insulin release. This combination of insulin resistance without augmented B-cell response may explain the hyperglycemia and other metabolic abnormalities seen in some protease inhibitor-treated patients. PMID- 11404535 TI - HIV testing among young adults and older adolescents in the setting of acute substance abuse treatment. AB - OBJECTIVES: To examine the feasibility and acceptance of HIV testing among youth in acute substance abuse treatment. METHODS: Youth, aged 18 to 25 years old, in state-funded inpatient detoxification completed a confidential demographic/risk behavior questionnaire, and were offered a choice of no testing, serum-based testing, or oral fluid (Orasure, Epitope, Inc., Beaverton, OR, U.S.A.) HIV testing. RESULTS: In all, 74% of 204 participants accepted HIV testing. In a multivariate model, female gender (odds ratio [OR], 0.32; 95% confidence interval [CI] 0.14-0.74) and having been recently tested (OR, 0.11; 95% CI, 0.05-0.26) were independently associated with refusing testing. Recent sexual activity (OR, 5.4; 95% CI, 1.5-20.4), recent use of methamphetamines (speed) or a combination of cocaine and heroin (speedball) (OR, 3.8; 95% CI, 1.6-9.0), and a recent perceived risk for HIV (OR, 4.6; 95% CI, 1.9-10.9) were independently associated with test acceptance. Thus, 150 of 150 (100%) chose the Orasure test. Overall, 64.6% (97 of 150) of those tested received their results, but among participants requiring a follow-up appointment to learn test results, only 9.2% (8 of 87) returned. CONCLUSIONS: HIV testing is feasible and acceptable in this population. All patients preferred Orasure testing to a serum enzyme-linked immunosorbent assay (ELISA). Most youth tested in detoxification will only learn their results if they are provided during treatment. Rapid HIV testing with same-day results could improve follow-up rates. PMID- 11404536 TI - Hospitalization rates in an urban cohort after the introduction of highly active antiretroviral therapy. AB - OBJECTIVE: Previous studies have shown a decrease in hospitalization rates associated with the introduction of highly active antiretroviral therapy (HAART). To evaluate hospitalization rates and patterns in discharge diagnoses that changed between 1995 and 1998 and to examine risk factors for hospitalization in HIV-positive patients, we conducted a cohort study. PATIENTS AND METHODS: All inpatient hospitalizations of 2,151 HIV-positive patients enrolled in our university-based HIV clinic between January 1, 1994 and December 31, 1998 with a CD4 count within a 6-month calendar semester were examined to evaluate hospitalization rates, discharge diagnoses, and intensive care department use. Negative binomial regression was used to assess the effect of various risk factors on hospitalization. RESULTS: Hospitalization rates decreased between 1995 and 1996 but increased between 1997 and 1998. In multivariate regression, female gender (incidence rate ratio [IRR], 1.45; p <.001), injection drug use (IRR, 1.36; p <.001), and having received no antiretroviral therapy were strong predictors of total hospitalization. White race, low CD4 count, and no antiretroviral treatment were strong predictors of hospitalization for an opportunistic infection. Female gender (IRR, 1.45; p <.001), African-American ethnicity (IRR, 1.22, p =.05), no antiretroviral treatment, and low CD4 counts were predictive of higher hospitalization rates for nonopportunistic infection related diagnoses. Intensive care department-use was associated with white ethnicity (IRR, 1.86; p =.028), heterosexual transmission of HIV (IRR, 1.90; p =.009), no antiretroviral treatment, and low CD4 count at enrollment. CONCLUSIONS: Our data indicate that hospitalization rates decreased between 1995 and 1997 after introduction of HAART, but that they then increased between 1997 and 1998, particularly for diagnosed nonopportunistic infections. If these trends continue, it indicates that patients may be developing previously unseen comorbidities and that HAART may have reached or exceeded a threshold in its effectiveness in reducing the clinical morbidity that results in hospital admission. PMID- 11404537 TI - Effect of ritonavir/saquinavir on stereoselective pharmacokinetics of methadone: results of AIDS Clinical Trials Group (ACTG) 401. AB - The effect of ritonavir 400 mg/saquinavir 400 mg twice daily on the stereoselective pharmacokinetics of methadone was examined in 12 HIV-infected, methadone-using study subjects. DESIGN: A 24-hour methadone pharmacokinetic study was performed before antiretroviral therapy was begun and after 15 days of therapy. Methadone concentration was measured by a chiral plasma assay because the drug is administered as a racemic mixture of R- and S-methadone, but only the R-isomer is active. Both changes in plasma protein binding and changes in objective and subjective opioid effect were monitored. RESULTS: Ritonavir/saquinavir administration was associated with 40% decrease in total S methadone AUC0-24hr and 32% decrease in R-methadone area under the curve (AUC)0 24hr, and both changes were statistically significant (p =.001 for both). When AUC was corrected for the changes in protein binding induced by ritonavir/saquinavir, R-methadone free AUC0-24hr decreased 19.6% whereas the S methadone decreased 24.6%, neither of these changes was statistically significant (p =.129 and p =.0537, respectively). This change in methadone exposure was not associated with any evidence of withdrawal from narcotics and no modification of methadone dose was required. CONCLUSIONS: Our data indicate that ritonavir/saquinavir administration is associated with induction of metabolism of methadone but this is greater for the inactive S-methadone. However, approximately 37% of the decrease in the total R-methadone exposure can be explained by protein binding displacement. Ritonavir/saquinavir can be used in HIV-infected people taking methadone without routine dose adjustments. PMID- 11404538 TI - Prognostic factors of combined viral load and CD4+ cell count responses under triple antiretroviral therapy, Aquitaine cohort, 1996-1998. AB - OBJECTIVE: To describe the viroimmunologic response and its prognostic factors 6 months after initiating triple antiretroviral therapy in a cohort of HIV-1 infected patients. METHODS: Positive virologic response during follow-up (VL+) was defined as plasma HIV RNA level <500 copies/ml and positive immunologic response (CD4+) as an increase of CD4+ count of at least 50 cells/mm3. Four categories of response were defined: VL+/CD4+; VL+/CD4-; VL-/CD4+ and VL-/CD4-. Prognostic factors were studied through a polytomous logistic regression (VL-/CD4 , as reference). RESULTS: Baseline characteristics of the 478 studied patients were: 22% at AIDS stage, 77% pretreated, median CD4+ cell count 195/mm3 and HIV RNA level 4.42 log. At 6 months 37.5% were VL+/CD4+; 15.7% VL+/CD4-; 23.8% VL /CD4+ and 23.0% VL-/CD4-. Baseline HIV RNA level was associated to a higher risk of VL-/CD4+ response. More advanced age was associated with a higher risk of isolated immunologic failure (VL+/CD4-), whereas pretreatment and saquinavir therapy were associated with a lower frequency of positive virologic response independently of immunologic response. CONCLUSION: HIV-RNA level, pretreatment, and saquinavir therapy were already known to be linked to therapeutic response. Based on our results, a high baseline HIV-RNA level is associated with isolated immunologic response; moreover, age should be of importance in treatment decision. PMID- 11404539 TI - Increase and plateau of CD4 T-cell counts in the 3(1/2) years after initiation of potent antiretroviral therapy. AB - We evaluated CD4 cell counts over a 3(1/2) year period following the initiation of potent antiretroviral therapy (ART) in the Multicenter AIDS Cohort Study. The study population included 314 HIV-infected gay men who provided CD4 cell counts for at least 2 years after the initiation of potent ART. Trends in CD4 cell counts and plasma HIV-RNA were analyzed by regression methods that incorporated the statistical dependencies of outcomes measured over time within individuals. Regardless of CD4 cell count at initiation of potent ART, CD4 cell counts increased significantly (p <.05) in the first 2 years after initiation. However, between 2 and 3(1/2) years after initiation, these counts neither increased nor decreased. The pattern of the proportion with plasma HIV-RNA <400 copies/ml was similar to CD4 cell count (i.e., increased significantly after initiation and plateau in the subsequent 1(1/2) years). The single most important predictor of the steady state CD4 cell count that was maintained between 2 and 3(1/2) years after initiation was the change in plasma HIV-RNA in the first year after initiation of potent ART. PMID- 11404540 TI - National trends in condom use among at-risk heterosexuals in the united states. AB - Based on national level surveys, we examined data relevant to the United States' overall effort to prevent the spread of HIV among heterosexual adults. We examined changes in condom use among at-risk heterosexuals over the past decade. The observed increases over time in condom use across all heterosexual at-risk population segments are consistent with the observed (declines) trends in HIV and syphilis in the 1990s. These results and findings from prior studies suggest that U.S. efforts to facilitate condom use and contain HIV and related sexually transmitted disease (STD)-cofactors among adult at risk heterosexuals was succeeding over most of the 1990s. The absence of national level behavioral trend data after 1996, and the ambiguities of HIV spread suggest some caution in projecting trends into this century. National and local efforts need to be directed at sustaining behavioral change and conducting more rigorous studies on population trends in HIV/STD-related behaviors/pathogens. PMID- 11404541 TI - Impact of pharmacy-based syringe access on injection practices among injecting drug users in Minnesota, 1998 to 1999. AB - In Minnesota, state legislation, enacted July 1998, provided for voluntary pharmacy sales of syringes/needles without a prescription for an accompanying drug. The goal was to stem HIV transmission among injecting drug users (IDUs) by providing greater access to sterile syringes. We used a pre/post evaluation design to investigate the impact of less restrictive syringe/possession laws on IDUs' HIV-related syringe practices. Independent cross-sectional samples of IDUs were recruited from street sites and a correctional facility immediately before and 1 year after enactment of the laws. Of the 671 IDUs interviewed, 570 (270 prelegislation and 300 postlegislation) had injected at least once in the 30 days before the interview. IDUs were more likely to purchase syringes at pharmacies after enactment of the laws (odds ratio [OR], 2.66; 95% confidence interval [CI], 1.83-3.85), yet did not change their behaviors regarding carrying unused syringes (OR, 0.90; 95% CI, 0.63-1.28). After adjusting for speedball injection and criminal history, syringe sharing decreased among IDUs (adjusted OR, 0.67; 95% CI, 0.45-1.00) yet syringe reuse remained the same (adjusted OR, 0.67; 95% CI, 0.40-1.11). Safe disposal of syringes did not differ significantly across the sampling periods (adjusted OR, 1.32; 95% CI, 0.84-2.06). Increased access to pharmacy syringes offers a first step at reducing HIV-related syringe practices but must be coupled with strong HIV prevention messages, drug treatment referrals, and safe syringe disposal options. PMID- 11404542 TI - Investigation of a secondary syringe exchange program for homeless young adult injection drug users in San Francisco, California, U.S.A. AB - This study investigated an HIV prevention program for homeless young adult injection drug users (IDUs) that combined a secondary syringe exchange program (SEP) with community-level activities. Homeless young IDUs were recruited from street-based settings in San Francisco, and a structured questionnaire was administered. The secondary SEP operated in a circumscribed geographic area, and for analytic purposes respondents were assigned to the intervention site group if they primarily spent time in this area (n = 67), or the comparison site group if they primarily spent time elsewhere (n = 55). Almost all (96%) intervention site youth had used the secondary SEP in the past 30 days and were significantly more likely to regularly use SEP. In bivariate analysis, comparison site IDUs were more likely to share syringes, reuse syringes, share the cotton used to filter drugs, and use condoms with casual sex partners only inconsistently. In multivariate analysis, comparison site remained positively associated with sharing syringes (adjusted odds ratio [AOR], 3.748; 95% confidence interval [CI], 1.406-9.988), reusing syringes (AOR, 2.769; 95% CI,1.120-6.847), and inconsistent condom use with casual sex partners (AOR, 4.825; 95% CI, 1.392- 16.721). This suggests that the intervention was effective in delivering SEP services to homeless young adult IDUs, and that IDUs who frequented the intervention site had a lower HIV risk than comparison group IDUs. PMID- 11404543 TI - HIV-1 incidence among male workers at a sugar estate in rural Malawi. AB - OBJECTIVES: To determine incidence of HIV and associated risk factors in two cohorts of men working at a sugar estate in rural Malawi. DESIGN: Prospective studies. METHODS: After counseling and obtaining informed consent, male workers were tested for HIV-1 and syphilis. Baseline HIV-seronegative men were enrolled in two follow-up studies in 1994 and 1998, and were retested for HIV and syphilis at 6-month follow-up visits. Demographic, behavioral, and medical history was collected at baseline. Cumulative HIV incidence based on Kaplan-Meier methods was estimated. HIV incidence was also estimated per 100 person-years (p-y). Crude and adjusted rate ratios for the association of risk factors with incident HIV infection were obtained using Cox proportional hazards models. RESULTS: HIV prevalence was 24.3% among 1692 men screened in 1994 and 21.0% among 1349 men screened in 1998 (p <.03). HIV incidence was extremely high during 1994 to 1995 (17.1% for that 1-year period). Incidence dramatically declined in 1996, averaging about 3.5% per year from 1996 through 1999. Among men enrolled in the 1998 cohort, HIV incidence during 1998 to 1999 was 3.8%. After controlling for potential confounders reactive syphilis was associated with a twofold risk of HIV acquisition in each cohort. CONCLUSIONS: Urgent preventive measures are needed to control the spread of HIV in this economically important occupational cohort. In addition to conventional educational messages to reduce risky sexual behavior, treatment of other sexually transmitted diseases should be considered. PMID- 11404544 TI - HIV transmission after suspension of highly active antiretroviral therapy. PMID- 11404545 TI - Pharmacokinetic parameters of protease inhibitors and the Cmin/IC50 ratio: call for consensus. PMID- 11404600 TI - Hermann von Helmholtz: a giant of science who cast his shadow on otolaryngology. PMID- 11404601 TI - Therapeutic efficacy of the Epley canalith repositioning maneuver. AB - OBJECTIVES/HYPOTHESES: The hypotheses of the current study are as follows: 1) That if the Epley canalith repositioning maneuver is an effective treatment for benign positional vertigo (BPV), relief from the vertigo should occur virtually immediately after the performance of the maneuver; 2) that the Epley canalith repositioning maneuver does provide almost immediate relief in BPV and should be the established treatment of choice for this disorder in both primary and tertiary care settings; and 3) that residual symptoms of lightheadedness and imbalance do persist after the resolution of the vertigo. The distinction of these symptoms from the vertigo is required for the accurate evaluation of the efficacy of positional maneuvers. STUDY DESIGN: Prospective cohort study in a tertiary care balance center. METHODS: Eighty-six patients (95 cases) with a history and physical examination consistent with active BPV were entered in the study. Patients were treated with a modified Epley canalith repositioning maneuver. A modified 360 degrees roll was used to treat those patients with horizontal canal BPV. Patients were provided with a preprinted diary in which they were to circle the answer most relevant to their symptoms for 14 days after the maneuver. Patients were then re-evaluated in the office at 2 weeks after the maneuver. RESULTS: The mean duration of the BPV before treatment was 9 weeks. Seventy-four percent of cases that were treated with one or two canalith repositioning maneuvers had a resolution of vertigo as a direct result of the maneuver. A resolution attributable to the first intervention was obtained in 70% of cases within 48 hours of the maneuver. An additional 14% of cases that were treated had a resolution of vertigo; however, it is not possible to say that these patients definitely benefited from the canalith repositioning maneuver. Only 4% of cases (three patients) manifested BPV that persisted after four treatments. Residual symptoms of lightheadedness or imbalance, or both, were frequent (47% of cases) but rarely required formal intervention with vestibular rehabilitation physical therapy. CONCLUSIONS: The Epley canalith repositioning maneuver results in a resolution of vertigo in the majority of patients (70% of cases) immediately after one treatment. It is safe and requires no special equipment or investigations. It should be established as the treatment of choice for BPV in both primary and tertiary care settings. PMID- 11404602 TI - Distortion-product otoacoustic emissions and cochlear microphonics: relationships in patients with and without endolymphatic hydrops. AB - OBJECTIVES: Because endolymphatic hydrops causes cochlear malfunction, and both otoacoustic emissions and cochlear microphonics measure specific cochlear activities, some insight into the pathology of Meniere's disease might be gained by using these two test modalities. Specifically, the involvement of cochlear outer hair cells in patients with endolymphatic hydrops may be detected. Furthermore, it is hoped that these two tests might help determine which regions of the cochlea are affected by endolymphatic hydrops, as well as where along the auditory pathway abnormalities are present. STUDY DESIGN: Data were gathered prospectively on patients presenting to a private, tertiary referral otology/neurotology practice. METHODS: From February 1999 to April 2000, clinical information was collected on patients presenting with vertigo, hearing loss (HL), sudden HL, fluctuant HL, aural fullness, and/or tinnitus. Data included demographics, diagnosis, pure-tone and speech audiometry, tympanometry, summating potential, action potential, cochlear microphonic, and distortion-product otoacoustic emissions. Descriptive statistics were calculated, and relationships between distortion-product otoacoustic emissions and cochlear microphonics in patients with hydrops (defined as summating potential to action potential ratio > or =0.40) and without hydrops were analyzed. RESULTS: Distortion-product otoacoustic emissions were present more often and had larger amplitudes at the lower frequencies. No differences were found in the presence of distortion product otoacoustic emissions across the frequencies for the two groups, but larger mean amplitudes were found for hydropic ears at 7966 Hz. As hearing levels worsened, both hydropic and nonhydropic ears were less likely to have emissions present; however, 18% of hydropic ears had emissions unexpectedly present when the pure-tone thresholds were > or =50 dB. The cochlear microphonic from the hydrops group tended to be smaller, but this was not statistically significant. Analysis of variance showed a small negative correlation between summating potential to action potential ratio and level of emission at 1968 Hz in hydropic ears; otherwise, there was no relationship between the ratio and emissions. The only statistically significant finding when analyzing the relationship between cochlear microphonic and otoacoustic emission was a small positive correlation between level of microphonic and level of emission at 1406 Hz in hydropic ears. No significant relationships were found between hearing thresholds and emissions or microphonics. CONCLUSIONS: Even though both distortion-product otoacoustic emissions (DPOAEs) and cochlear microphonics (CMs) measure specific cochlear activities, they were not found to be useful for differentiating patients with hydrops from those without. In some patients, however, unexpected distortion product otoacoustic emissions were present. This may represent localizing information about which regions of the cochlea are being affected by hydrops in these patients. Also, a small positive correlation between the CM and the DPOAE at 1406 Hz was detected in the hydrops group, which may represent the effects of endolymphatic hydrops on the outer hair cell. Future investigations involving hydropic patients with unexpected DPOAEs and studies looking for more DPOAE and CM correlations at frequencies surrounding 1406 Hz are being planned. PMID- 11404603 TI - Results of the bone-anchored hearing aid in unilateral hearing loss. AB - OBJECTIVES: The advantages of binaural hearing are well established and universally accepted. However, a tendency remains to withhold the benefits of binaural hearing to adults and children with one normal ear. The purpose of this study is to demonstrate the benefit of the bone-anchored hearing aid (BAHA) in a group of patients with unilateral conductive or mixed hearing loss. STUDY DESIGN: This is a prospective study of nine patients (five males and four female patients) with conductive or mixed hearing loss who met the criteria for BAHA except for having normal hearing in the other ear. They had congenital aural atresia or mastoidectomies secondary to chronic ear infections with or without cholesteatoma or had a temporal bone tumor excised METHODS: Patients had evaluations before and after implantation, including audiological testing and responses to a standardized hearing handicap questionnaire. Statistical analyses of the data were made using the Wilcoxon signed rank test and the paired Student t test for repeated measures. RESULTS: All patients had tonal and spondee threshold improvement with BAHA when compared with thresholds before treatment. Speech recognition performance in BAHA-aided conditions was comparable to the patient's best score in unaided condition. Patients reported a significant improvement in their hearing handicap scores with the BAHA. CONCLUSIONS: The use of BAHA has significantly improved the hearing handicap scores in patients with unilateral conductive or mixed hearing loss. The proven safety and efficacy of the device promote its use in unilateral cases that traditionally had been left unaided. PMID- 11404604 TI - Does high-frequency pseudo-random rotational chair testing increase the diagnostic yield of the eng caloric test in detecting bilateral vestibular loss in the dizzy patient? AB - OBJECTIVES: To assess the incremental diagnostic yield of testing vestibulo ocular (VOR) gain with high-frequency pseudo-random rotational chair (PsRRC) over testing with bithermal electronystagmography caloric tests in the dizzy patient, particularly in detecting bilateral vestibular loss. PATIENTS AND METHODS: One hundred ninety-eight patients presenting with dizziness underwent PsRRC and caloric testing. The VOR gain on PsRRC was measured at 0.32 to 5.0 Hz, with gain categorized as normal or decreased. PsRRC results were compared with caloric responses, also categorized as normal, or into graded categories of unilateral or bilateral vestibular loss. RESULTS: Reduced PsRRC gain was found in 29 (15%) patients, and reduced caloric tests responses in 70 (35%), with 25 (13%) having bilateral loss. Of patients with reduced chair gain, 25 of 29 (86%) demonstrated bilateral caloric loss. PsRRC gain was normal in most patients with unilateral caloric weakness, but was decreased in all patients with bilateral caloric weakness. The probability of a patient with completely normal caloric responses having an abnormal rotation chair in this study group was under 1% (1 of 128). CONCLUSIONS: PsRRC testing does not offer much additional diagnostic benefit when caloric responses are normal. It is useful in specific conditions, such as unilateral caloric loss for which the patient is not compensating, borderline caloric loss when traditional water caloric tests cannot be used, or for monitoring progressive bilateral vestibular loss. PMID- 11404605 TI - A stepwise surgical technique using the medial orbital floor as the key landmark in performing endoscopic sinus surgery. AB - HYPOTHESIS: The medial orbital floor (MOF) and adjacent bony ridge of the antrostomy, when combined with columellar measurements, are easily identifiable and consistent anatomic reference points from which critical orbital and skull base structures can be found during endoscopic sinus surgery. METHODS: Two examiners, with varying endoscopic sinus surgery experience, performed endoscopic and direct measurements from the columnella and medial orbital floor to critical orbital and skull base structures on 11 human cadaver heads (18 sides). The distances to four critical skull base or orbital structures and to the anterior and posterior wall of the sphenoid sinus were measured. The mean, ranges, and standard deviations for all measurements (endoscopic and direct) were calculated and simple regression analysis was performed. RESULTS: The mean and range of values for each of the variables correlated well between examiners, and between endoscopic and direct measurements. There was slightly more variability in measurements when the MOF was used. However, the differences were no more than a few millimeters and did not appear to affect the overall clinical use of these values. CONCLUSIONS: The MOF and adjacent bony ridge of the antrostomy, when combined with columellar measurements, are easily identifiable and consistent anatomic landmarks that are not affected by the presence of significant inflammatory disease or previous surgery. These reference points provide even the most inexperienced surgeon with precise anatomic localization within the paranasal sinuses. They also determine the correct anteroposterior trajectory into the sphenoid sinus, whereby inadvertent intracranial or intraorbital complications may be avoided. PMID- 11404606 TI - The silent sinus syndrome: a case series and literature review. AB - PURPOSE: The purpose of this study is to describe the clinical and pathologic features of a form of chronic maxillary atelectasis referred to as the silent sinus syndrome, which is characterized by progressive enophthalmos secondary to maxillary collapse resulting from maxillary sinus hypoventilation. METHODS: A retrospective medical record analysis was carried out to identify patients with enophthalmos secondary to maxillary collapse. Clinical records, including ophthalmology and otolaryngology evaluations as well as computed tomography scans and operative reports, were carefully examined. A complete literature review for relevant studies was performed to examine possible pathophysiology and similar cases. RESULTS: Four patients with enophthalmos and asymptomatic maxillary sinus disease were identified. On computed tomography, all four of the patients had opacified, partially collapsed maxillary sinuses with osteopenia of the sinus walls and orbital floor displacement resulting in enophthalmos. All four underwent successful functional endoscopic sinus surgery and transconjunctival orbital floor repair. CONCLUSION: In some instances, chronic maxillary atelectasis can present with enophthalmos secondary to collapse of the maxillary sinus. For reasons that are unclear, the sinus component of the disease remains asymptomatic and is discovered only after thorough evaluation of the enophthalmos. PMID- 11404607 TI - Laryngopharyngeal reflux symptoms improve before changes in physical findings. AB - BACKGROUND: Patients with laryngopharyngeal reflux (LPR) undergoing treatment appear to have improvement in symptoms before the complete resolution of the laryngeal findings. OBJECTIVE: To determine whether patients with LPR experience an improvement in symptoms before the complete resolution of the laryngeal findings. METHODOLOGY: Forty consecutive patients with LPR documented by double probe pH monitoring were evaluated prospectively. Symptom response to therapy with proton pump inhibitors was assessed at 2, 4, and 6 months of treatment with a self-administered reflux symptom index (RSI). In addition, transnasal fiberoptic laryngoscopy (TFL) was performed and a reflux finding score (RFS) was determined for each patient at each visit. RESULTS: The mean RSI at entry was 19.3 (+/- 8.9 standard deviation) and it improved to 13.9 (+/- 8.8) at 2 months of treatment (P <.05). No further significant improvement was noted at 4 months (13.1 +/- 9.8) or 6 months (12.2 +/- 8.1) of treatment. The RFS at entry was 11.5 (+/- 5.2), and it improved to 9.4 (+/- 4.7) at 2 months, 7.3 (+/- 5.5) at 4 months, and 6.1 (+/- 5.2) after 6 months of treatment (P <.05 with trend). CONCLUSIONS: Symptoms of LPR improve over 2 months of therapy. No significant improvement in symptoms occurs after 2 months. This preliminary report demonstrates that the physical findings of LPR resolve more slowly than the symptoms and this continues throughout at least 6 months of treatment. These data imply that the physical findings of LPR are not always associated with patient symptoms, and that treatment should continue for a minimum of 6 months or until complete resolution of the physical findings. PMID- 11404608 TI - Antisense cyclin D1 enhances sensitivity of head and neck cancer cells to cisplatin. AB - OBJECTIVES: Cyclin D1 is a cell cycle regulatory factor that modulates a critical step in cell cycle control. Cyclin D1 is overexpressed in a significant proportion of head and neck cancers and correlates with a poor prognosis. Abrogation of cyclin D1 action through antisense cyclin D1 shows promise as an antitumor therapy, with an inhibitory effect in head and neck squamous cell carcinoma both in vitro and in vivo. The suppressive effect of antisense cyclin D1 in head and neck cancer xenografts in nude mice is incomplete, however, suggesting that combination with another antitumor agent is necessary for complete tumor eradication. Cisplatin is a widely used chemotherapeutic agent in head and neck cancer, and is particularly effective in combination with radiation therapy. In this study, we investigate whether antisense cyclin D1 enhances the sensitivity of head and neck cancer cells to cisplatin. Such an enhancement of sensitivity would suggest that combination therapy using antisense cyclin D1 and cisplatin would be an effective treatment modality for head and neck cancer. STUDY DESIGN: Antisense cyclin D1 was transfected into the head and neck squamous cell carcinoma cell line CCL23 using a plasmid vector. Both the parental CCL23 cells and the antisense cyclin D1-transfected CCL23 cells (CCL23AS) were treated with cisplatin at increasing concentrations. The dosage of cisplatin ranged from 1 microg/mL to 10 microg/mL. Initial exposure to cisplatin was for 2 hours, with increasing exposure times in succeeding experiments. Cell viability assays were done following cisplatin exposure. Dose response curves for the two cell lines were plotted and compared. Western blot analyses were done on the cisplatin treated cell lines to determine levels of cyclin D1 expression. RESULTS: Increasing concentrations of cisplatin resulted in significantly higher rates of cell killing in the antisense cyclin D1-transfected cells than in the parental cells. The ID50 values for the parental CCL23 cells and the antisense cyclin D1 transfected CCL23 cells were 7 microg/mL and 3 microg/mL, respectively, indicating significant enhancement of sensitivity to cisplatin in the antisense cyclin D1-transfected cells. Western blot analyses demonstrated decreased expression of cyclin D1 in the CCL23AS cells with increasing doses of cisplatin, compared with the parental CCL23 cells. CONCLUSIONS: Antisense cyclin D1 transfected CCL23 cells demonstrate an enhanced sensitivity to the effects of cisplatin compared with the parental cell line. Although the mechanism for this phenomenon is not completely understood, the data suggests the potential use of combination therapy using antisense cyclin D1 and cisplatin for head and neck cancers. While neither agent alone can completely eradicate head and neck cancers, the synergistic effect of the two may be an effective therapeutic protocol for refractory head and neck cancers. Future investigation into the combination of antisense cyclin D1 with cisplatin for treatment of head and neck cancer is needed. PMID- 11404609 TI - Cause of death in patients with well-differentiated thyroid carcinoma. AB - OBJECTIVES: The aim of this study is to examine in detail patients dying of well differentiated thyroid carcinoma. STUDY DESIGN: A retrospective chart review with follow-up. METHODS: Data were collected from 522 consecutive cases of differentiated thyroid carcinoma treated by one endocrinologist and four surgeons at Mount Sinai Hospital, Toronto, Ontario, Canada, from 1964 to 1999. RESULTS: Ten patients died as a direct result of thyroid carcinoma; 19 other deaths were unrelated. Five of 102 patients were men (5%) and 5 of 420 were women (1%); the median age at diagnosis was 68.5 years (range, 49-82 y). No cases were stage I; three, stage II; two, stage III; and five, stage IV. Pathologically papillary carcinoma was found in six of the patients who died, follicular carcinoma in three patients, and Hurtle cell carcinoma in one patient. The causes of death were local invasion or compression of the trachea in two cases and distant metastases in eight patients. Median survival was 3.5 years (range, 1 mo-20 y). CONCLUSIONS: All patients dying of well-differentiated thyroid carcinoma had neck nodes, extrathyroidal spread, or distant metastases at presentation and were older than 49 years of age. Many presented because of their distant metastases. Death resulting from local disease was unusual, with most patients dying of distant metastases. PMID- 11404610 TI - Hyperthermic, supersaturated humidification in the treatment of xerostomia. AB - OBJECTIVES: To investigate the role of hyperthermic, supersaturated humidification in the treatment of radiation-induced xerostomia. STUDY DESIGN: A randomized, controlled, crossover pilot study of patients with symptomatic xerostomia following radiotherapy for head and neck cancer. Patients compared a standard bedside humidifier with a new device delivering hyperthermic, supersaturated humidification through a nasal cannula. METHODS: The patients were randomized to a 2-week course of standard, cool air bedside humidification or to hyperthermic, supersaturated humidification through a nasal cannula (Vapotherm Inc., Annapolis, MD). After a 1-week washout period, patients were crossed over to the opposite device for another 2 weeks. The patients underwent physical examination initially and after each trial period using an objective xerostomia scale, and then completed a questionnaire quantifying their symptoms. Patients additionally rated their symptoms at home, twice daily, using a visual analogue scale. RESULTS: Using the Oral Assessment Guide, lip scores went from 1.67 initially to 1.67 after control and 1.67 after Vapotherm. Tongue scores were 1.67, 1.5, and 1.83, respectively. Saliva scores were 1.67, 1.5, and 1.5, respectively. Mucous membranes scores were 1.5, 1.67, and 1.5, respectively. The physical examination scores at these four sites were not significantly different between control and Vapotherm (P =.78,.78,.72, and.37, respectively). The patient symptom questionnaire and visual analogue scores also revealed no significant difference between the two devices. CONCLUSION: The Vapotherm MT-3000 device appears to provide minimal or no additional relief from radiation xerostomia over standard bedside humidifiers. Further investigation may be warranted with newer models of the device and with disorders of mucociliary clearance. PMID- 11404611 TI - Adenotonsillar hypertrophy and Epstein-Barr virus in pediatric organ transplant recipients. AB - OBJECTIVES/HYPOTHESIS: Epstein-Barr virus-related (EBV-related) lymphoid hyperplasia of the tonsils and adenoids is a precursor to post-transplantation lymphoproliferative disorder (PTLD). The incidence of post-transplantation adenotonsillar hypertrophy, a potential early sign of PTLD or EBV-related lymphoid hyperplasia, is not known. We sought to identify potential risk factors for adenotonsillar hypertrophy manifested as EBV-related hyperplasia and early PTLD in the pediatric solid organ transplant population. STUDY DESIGN: Cross sectional analysis. METHODS: We developed a 65-point questionnaire concerning obstructive sleep disorder and upper respiratory tract infections and an 8-point focused physical examination, to identify prevalence of and risk factors for adenotonsillar hypertrophy in the pediatric transplant population. We evaluated 120 pediatric solid organ transplant recipients by parental questionnaire and focused adenotonsillar physical examination. RESULTS: Of the 120 patients, 62 had undergone liver transplantation and 58 had undergone kidney transplantation. Overall, the mean questionnaire score was 8.36 (range, 0-40) and the mean physical examination score was 3.86 (range, 1-8). Patients whose EBV serological test result was negative at the time of transplant had higher scores for both the questionnaire (mean score, 10.24) and the physical examination (mean score, 4.56) than those whose EBV serological test result was positive at the time of transplantation (scores of 7.38 and 3.30 for questionnaire and physical examination, respectively). The difference in examination scores was statistically significant (P <.003). CONCLUSIONS: Epstein-Barr virus seronegativity at the time of organ transplantation is a known risk factor for PTLD, with associated risk of developing EBV-related lymphoid hyperplasia. Our results indicate a higher incidence of symptoms and signs consistent with adenotonsillar hypertrophy in the EBV seronegative population. Adenotonsillar hypertrophy may be a precursor to EBV-related lymphoid hyperplasia and PTLD and must be identified in this patient population. PMID- 11404612 TI - Histological characterization of the thyroglossal tract: implications for surgical management. AB - OBJECTIVES: In the current report, we characterized the relationship between the central hyoid bone and the thyroglossal tract and determined the prevalence of ectopic thyroid follicles in the adjacent soft tissues. STUDY DESIGN: Retrospective pathological analysis. METHODS: The resected specimens from 104 patients who underwent a modified Sistrunk procedure with wide-field dissection were retrospectively analyzed. Under light microscopy, serial sections were examined to determine whether the thyroglossal tract passed anterior to, posterior to, or within the hyoid bone. Specimens were also examined for the presence of thyroid follicles in the periductal and pericystic soft tissues. RESULTS: In 50 cases (48%), the tract position in relation to the hyoid bone could not be identified secondary to extensive arborization, marked inflammation, specimen fragmentation or a combination of these. Thyroid follicles were observed in 9 (18%) of these specimens. In the remaining 54 cases (52%) the tract was located anterior to the central arch of the hyoid bone in 39 (72%) and posterior to it in 15 (28%). Thyroid tissue was observed in 46% of specimens (P =.004), regardless of whether the tract was anterior or posterior. CONCLUSIONS: These results demonstrate that tract position often cannot be defined, but when a portion is dominant, it is likely to be anterior to the hyoid bone. Ectopic thyroid tissue can be found in almost 50% of specimens when the tract position is identifiable. With appropriate surgical management, a recurrence rate of less than 4% can be expected, despite the presence of ductule multiplicity, marked inflammation, tract position posterior to the hyoid bone, and ectopic thyroid follicles. PMID- 11404613 TI - Allergic fungal rhinosinusitis: current theories and management strategies. AB - The combination of nasal polyposis, crust formation, and sinus cultures yielding Aspergillus was first noted in 1976 by Safirstein,1 who observed the clinical similarity that this constellation of findings shared with allergic bronchopulmonary Aspergillosis (ABPA). Eventually this disease came to be known as allergic fungal rhinosinusitis (AFS). As clinical evidence of AFS accumulated, controversy regarding its etiology, pathogenesis, natural history, and appropriate treatment naturally emerged. Despite past and current efforts, many of these controversies remain incompletely resolved, but continuing clinical study has illuminated some aspects of the disease and has led to an improved understanding of AFS and its treatment. Fungi associated with the development of AFS are ubiquitous and predominantly of the dematiaceous family. The eosinophilic host response to the presence of these fungi within the nose and paranasal sinuses gives rise to those clinical manifestations of the disease (nasal polyps, expansile mucocele formation, allergic fungal mucin, etc.). Exposure alone to these fungi, however, appears to be insufficient to initiate the disease. At the present time it is likely that initiation of the inflammatory cascade leading to AFS is a multifactorial event, requiring the simultaneous occurrence of such things as IgE-mediated sensitivity (atopy), specific T-cell HLA receptor expression, exposure to specific fungi, and aberration of local mucosal defense mechanisms. A variety of treatment plans for AFS have emerged, but the potential for recidivism remains well recognized, ranging from 10% to nearly 100%, suggesting the need for continued study of this disease and fueling present controversy. This article is intended to review current data and theories regarding the pathophysiology of AFS, as well as the role of various surgical and nonsurgical forms of therapy. PMID- 11404614 TI - An interpretation of the structural changes responsible for the chronicity of rhinoscleroma. AB - OBJECTIVE: To identify the structural changes of the proliferative phase of rhinoscleroma which could be responsible for the chronicity of the disease. STUDY DESIGN: Observational research. METHODS: Samples of friable tissue taken from the nasal mucosa of nine untreated patients were processed for light and ultrastructural microscopy. RESULTS: The majority of changes contributing to the chronicity of the disease occurred in the subepithelium and followed three closely related but distinct events. In the first (infiltrative), subepithelial invasion by the Klebsiella was followed by its active multiplication and proliferation of capillaries. In the second (neutrophilic), large numbers of neutrophils were delivered into this space. Neutrophils actively phagocytized the Klebsiella but appeared to die at an accelerated rate without completing digestion of the microorganisms. In the third event (histiocytic), histiocytes entered the subepithelium and engaged in unrestrained phagocytosis of decaying neutrophils, Klebsiella, and debris. During this process, the histiocytes' phagosomes underwent massive dilation, thus becoming Mikulicz cells. Mikulicz cells were unable to consistently destroy the Klebsiella and eventually ruptured, releasing them into the interstitium. Evidence was found that an autophagic process might contribute to phagosome distention and to the rupture of the vacuolar membranes and cell wall. CONCLUSIONS: Several critical changes responsible for the chronicity of rhinoscleroma occur during the proliferative phase of the disease. The majority of these take place in the subepithelium and include: 1) factors leading to the transformation of histiocytes into Mikulicz cells, 2) the inability of these cells to consistently destroy the Klebsiella, 3) their rupture releasing viable Klebsiella, and 4) the intrinsic resistance of the pathogen. PMID- 11404615 TI - Endoscopic extraction of an intranasal tooth: a review of 13 Cases. AB - OBJECTIVE: To present the clinical experience of a series of 13 consecutive cases of an intranasal tooth treated by endoscopic extraction during a 15-year period. STUDY DESIGN: Retrospective review. METHODS: The records of 13 patients who underwent endoscopic extraction of an intranasal tooth at Taipei Medical University Hospital and Chang Gung Memorial Hospital between 1986 and 2000 were reviewed. RESULTS: All the patients had an uneventful recovery and the presenting symptoms were completely relieved after surgery. The follow-up period was 6 months to 14 years 5 months (average, 9 yr 2 mo). Examination of these extracted teeth showed that 11 were supernumerary teeth and two were ectopic permanent canine teeth. CONCLUSIONS: Endoscopic extraction of the intranasal tooth has the advantages of good illumination, clear visualization, and precise dissection, and in our experience the result of endoscopic extraction of an intranasal tooth is satisfactory. We recommend the routine use of a rigid endoscope in the treatment of an intranasal tooth. PMID- 11404616 TI - Expression patterns of cytokeratins in retraction pocket cholesteatomas. AB - OBJECTIVES: To investigate the patterns of cytokeratin (CK) expression in retraction pocket cholesteatoma. STUDY DESIGN: An animal model study. METHODS: Retraction pocket cholesteatomas were induced by electrocautery of the eustachian tube orifice in 24 mongolian gerbils. They were divided into normal and cholesteatoma groups of clinical stages I to IV. The antibodies to pan cytokeratin CK 1/10, CK 5/6, CK 4, and CK 13/16 were used for immunohistochemical staining. The intensity of staining in each group as measured with densitometry was compared regarding anatomical sites and clinical stages. RESULTS: In retraction pocket cholesteatoma, CK expression was altered only at focal sites such as the pars tensa of the tympanic membrane. The change of CK expression was observed only at certain stages of cholesteatoma formation. In keratinocytes from cholesteatomas, CK 13/16 was overexpressed compared with control specimens, indicating hyperproliferation. The site with the most prominent change in retraction pocket cholesteatoma was somewhat different from that in canal ligation cholesteatoma in a previous study. CONCLUSIONS: The results suggested that aural cholesteatoma is a disease with a spectrum of pathological conditions and that the transmigration and hyperproliferation process of squamous epithelium occurs in areas adjacent to the cholesteatoma. PMID- 11404617 TI - Cycline-dependent kinase inhibitor, p27 (KIP1), is associated with cholesteatoma. AB - OBJECTIVE: Cyclin-dependent kinases (CDKis) can arrest the cell cycle, which in turn inhibits the cell proliferation. P27 (KIP1) is a CDKi and acts as a tumor suppressor gene. In this study, we aimed to investigate the role of p27 CDKi in cholesteatoma, a disease characterized by the presence of hyperproliferative squamous epithelium. STUDY DESIGN: Immunohistochemical staining of 15 cholesteatoma and 18 control ear canal skin samples, which were taken intraoperatively, was performed for p27 positivity. METHODS: The monoclonal antibodies to p27 were used for immunohistochemical staining of the sections. The streptavidin-biotin horseradish method was used. The number of cells staining positive for p27 was calculated, and the intensity of p27 positivity was graded. RESULTS: P27 positivity was obtained in 9 (50%) of 18 skin tissues. In the cholesteatoma tissues, p27 positivity was found only in 2 (13.3%) tissue samples. The difference between the groups were statistically significant (P =.03). The mean numbers of p27 positivity were 11.8 +/- 15.5 and 1.4 +/- 3.8 (mean +/- standard deviation) in the skin and cholesteatoma samples, respectively. This difference was also statistically significant (P =.02). The p27 results of primary and secondary cholesteatoma samples were not significantly different (P =.3). The results of p27 were not related to the gender of the patients (P =.8). CONCLUSION: P27 is involved directly or indirectly in the occurrence of cholesteatoma. Alterations of p27 levels in keratinocytes can influence the proliferative state of the keratinocytes. Altered p27 levels in cholesteatoma may suggest a molecular pathology in cholesteatoma. The search for significance of CDKis seems promising to better understand the pathogenesis of cholesteatoma. PMID- 11404618 TI - The levels of expression of galectin-3, but not of galectin-1 and galectin-8, correlate with apoptosis in human cholesteatomas. AB - OBJECTIVES: To investigate whether galectins 1, 3, and 8 are expressed in human cholesteatomas and whether any such expression does correlate with the level of apoptosis, which is, as we have previously shown, predictive of recurrence.7 STUDY DESIGN: The analysis of 52 cholesteatomas resected by the same surgeon by means of canal wall up and canal wall down procedures. METHODS: The immunohistochemical levels of expression of galectins 1, 3, and 8 were quantitatively determined (using computer-assisted microscopy) on conventional histological slides by means of specific anti-galectin-1, anti-galectin-3, and anti-galectin-8 antibodies. The level of apoptosis in each cholesteatoma under study had already been determined 7 by means of the in situ labeling of nuclear DNA fragmentation (Tolt-mediated dUTP nick end labeling [TUNEL] staining). RESULTS: Galectin-1 was expressed markedly in both the epithelial and the connective tissue areas of all the cholesteatomas under study. The levels of expression of galectin-3 and galectin-8 were considerably lower than that of galectin-1. The level of expression of galectin-3 correlated both highly and positively with the level of apoptosis. CONCLUSIONS: An upregulation of galectin 3 (known to have an antiapoptotic and antianoikis effect in certain model systems) expression, which is associated with pronounced apoptotic activity, could have a physiologically protective effect against the characteristically substantial apoptotic features occurring in recurrent cholesteatomas. PMID- 11404619 TI - Three-dimensional image for the middle fossa approach and its anatomical considerations. AB - OBJECTIVE: To investigate the usefulness and significance of three-dimensional images in the middle cranial fossa approach. STUDY DESIGN: Retrospective study. METHOD: Three-dimensional images of 12 patients with acoustic neuromas were made for surgical guidance. An image showing an ideal surgical view was prepared, which showed no overlap between the labyrinthine bone and internal auditory canal (IAC) with a slight gap between them. The IAC was located and opened referring to this image. RESULTS: Directions of the images in the ideal surgical view were anterior from lateral position in all cases. Viewing toward the vestibule, angle from lateral plane was 24 degrees +/- 12 degrees (mean +/- standard deviation). In 11 of 12 patients, the IAC was safely opened; in one patient the IAC could not be opened because of bleeding. Overall hearing preservation rate was 82%. CONCLUSION: Our method facilitated not only in the identification of the IAC but also in avoiding inner ear damage. Our data indicated that the center of craniotomy should be made anterior to the external auditory canal and surgery should be viewed from an anterolateral position for a safe procedure. PMID- 11404620 TI - Benign positional vertigo without detectable nystagmus. AB - OBJECTIVES: To demonstrate that for treatment of benign positional vertigo it is not necessary to identify a positional nystagmus. STUDY DESIGN: Prospective trial of 43 patients with positional vertigo without clinical evidence of positional nystagmus who were treated with a modified canal-repositioning maneuver. METHODS: Results were compared with the results obtained in the treatment of posterior semicircular cupolo-canalithiasis with typical positional nystagmus with the same repositioning maneuver. RESULTS: Treatment of 43 patients with positional vertigo without positional nystagmus resulted in a 60.46% (26/43) complete recovery rate and a 6% (3/43) persistence of disorder rate compared with a 90% complete recovery rate obtained in 90 patients with typical benign positional paroxysmal vertigo (with positional nystagmus). CONCLUSIONS: What is attempted by this work is to confirm that through clinical evaluation based on history and positioning tests alone, one can obtain acceptable results in treatment of supposed canalithiasis without having detected the pathognomonic nystagmus. PMID- 11404621 TI - Selective vestibular ablation by KTP laser in endolymphatic hydrops. AB - OBJECTIVES AND HYPOTHESIS: Vertigo, the cause of disability in many patients with Meniere's disease, may be the result of the effects of endolymphatic hydrops on the semicircular canals. We hypothesize that intractable vertigo may be controlled by destruction of the semicircular canal neuroepithelium using visible light lasers without the need for extensive fenestration of the bony labyrinth. This study was designed to assess the cochlear effects of potassium titanyl phosphate (KTP) laser-assisted triple semicircular canal ablation (TSCA) in endolymphatic hydrops. STUDY DESIGN: Randomized, prospective, and controlled. METHODS: Forty-one adult guinea pigs underwent either a unilateral endolymphatic duct occlusion to induce hydrops or a sham procedure. Ten weeks after induction of the hydrops, a KTP laser-assisted TSCA or a sham surgery was performed. RESULTS: Electrocochleographic thresholds to clicks and tone-bursts (2-20 kHz) did not change significantly up to 4 weeks after TSCA in hydropic ears. Cross sectional histology confirmed the presence of hydrops and the ablation of the semicircular canals. Cochlear whole-mounts for hair cell counts showed no significant loss of outer or inner hair cells in hydropic ears treated with TSCA. CONCLUSION: KTP laser-assisted TSCA can be performed in the guinea pig model of endolymphatic hydrops without significant loss of hearing. Evaluation of this technique may be warranted in patients with intractable Meniere's disease. PMID- 11404622 TI - DiHA (dextranomers in hyaluronan) injections for treatment of insufficient closure of the vocal folds: early clinical experiences. AB - OBJECTIVE: To determine the clinical effects of injecting DiHA (a mixture of dextranomer molecules and 1% hyaluronan solution in equal proportions) as a "space filler" into the vocal folds of patients experiencing insufficient closure of the vocal folds resulting from either unilateral vocal fold paralysis or bowed vocal folds. STUDY DESIGN: A consecutive series of patients with insufficient closure of the vocal folds treated with DiHA injections and prospectively followed up with videostroboscopy and voice recordings that were evaluated by expert panels. METHODS: During a 21/2-year period, 14 patients with insufficient closure of the vocal folds were included. Six had a unilateral vocal fold palsy and 8 patients had "bowed" vocal folds. DiHA was injected into one vocal fold. Laryngoscopic examination and voice recordings were performed pre- and postoperatively. Expert panels did the evaluation of the stroboscopy examination and the perceptual voice analysis. RESULTS: Three patients were operated on with local and 11 under general anesthesia. All patients with unilateral palsy improved their stroboscopic status regarding both the wave and the closure after injection. Also, their voice parameters were significantly improved. Among the patients with bowed vocal folds, all except 2 showed an improvement in the glottic wave, 5 of 8 improved in glottic closure, and 3 improved their voice parameters significantly. CONCLUSION: This early clinical experience indicates that DiHA fulfills most of the requirements of an ideal "space-filling" substance for voice restoration in patients experiencing insufficient vocal fold closure. PMID- 11404623 TI - Histology of injected autologous fascia in the paralyzed canine vocal fold. AB - OBJECTIVE: To evaluate the histology of minced and injected autologous fascia graft in the augmentation of unilateral vocal fold paralysis. STUDY DESIGN: Prospective study using a canine model. METHODS: Nine dogs were operated. At first, a piece of fascia was harvested from fascia lata and minced into tiny chips with a scalpel. Cutting off a section of the recurrent nerve paralyzed the right vocal fold. The minced fascia-paste (0.1 mL) was injected using a pressure syringe into the paralyzed thyroarytenoid muscle under direct laryngoscopy. Two animals were killed at 3 days, one at 10 days, three at 6 months, and three at 12 months postinjection. Each dog underwent laryngectomy and serial coronal sections of paraffin blocks from the posterior part of the vocal folds were made. RESULTS: The dogs experienced no complications perioperatively or during follow-up. Under microscopy, muscle of the paralyzed vocal fold was atrophied in comparison to the contralateral control. There was an acute inflammatory reaction induced by the graft. This did not exist in the specimens taken at 6 and 12 months. No extensive edema, areas of necrosis, or formation of granulomas was seen at any time. Maturation of the graft was characterized by active collagen remodeling up to 12 months. At that time the graft consisted of firm, condensed fibrous tissue. Scar formation around the graft was moderate, and the subepithelial layer of the vocal fold remained undisturbed. Each graft consisted of singular foreign bodies from the polyamide mincing plate. We cannot exclude that their presence would have had an impact on the final architecture of the graft. CONCLUSION: In a canine vocal fold, the free fascia graft is well tolerated and after 12 months a well organized, collagen rich tissue is seen on histological sections. The findings are in accordance with clinical studies applying free fascia grafts. PMID- 11404624 TI - Uvulopalatopharyngoplasty, maxillomandibular advancement, and the velopharynx. AB - OBJECTIVE: To evaluate the presence of velopharyngeal insufficiency (VPI) symptoms and the associated changes of the velopharyngeal anatomy in patients who underwent maxillomandibular advancement (MMA) for persistent obstructive sleep apnea (OSA) after uvulopalatopharyngoplasty (UPPP). METHODS: Preoperative and postoperative cephalometric radiographs were analyzed to assess the anatomic changes of the velopharynx. In addition, a questionnaire survey was sent to the patients between 6 to 12 months after MMA. The questionnaires evaluated the presence and extent of VPI symptoms, including nasal regurgitation while eating or drinking as well as hypernasal speech. A 10-cm visual analog scale (VAS 0-10) was included to assess the impact of VPI symptoms on the patient's quality of life. In the patients who reported VPI symptoms, telephone interviews were conducted 1 year after the survey to evaluate the changes in VPI symptoms over time. RESULTS: Fifty-two of the 65 questionnaires were returned. Five patients (9.6%) reported nasal regurgitation of liquids when drinking hastily, with 2 patients reporting the occurrences as occasional and 3 patients reporting as rare. The impact of these symptoms on the patient's quality of life was minimal (VAS 0.6 +/- 0.4). Regurgitation of food or hypernasal speech was not reported. The telephone interviews 1 year later revealed that the symptoms have completely resolved in all 5 patients. Comparison of the preoperative and postoperative cephalometric radiographs demonstrated the pharyngeal depth increase was 48% of the amount of maxillary advancement and the functional pharyngeal length increased 53% of the maxillary advancement. The functional depth of the pharynx after MMA was significantly greater in the patients with VPI symptoms (P=.01). CONCLUSION: The results of this study suggest that patients who undergo MMA for persistent OSA after UPPP have a low risk of developing VPI. If symptoms occur postoperatively, they are mild and have minimal effect on the patient's quality of life; moreover, the symptoms usually resolve over time. PMID- 11404625 TI - Delayed regional metastases, distant metastases, and second primary malignancies in squamous cell carcinomas of the larynx and hypopharynx. AB - OBJECTIVE: To determine the impact of delayed regional metastases, distant metastases, and second primary tumors on the therapeutic outcomes in squamous cell carcinomas of the larynx and hypopharynx. STUDY DESIGN: Chart review and statistical analysis. METHODS: A retrospective tumor registry analysis was made of patients with squamous cell carcinomas of the larynx and hypopharynx who were treated with curative intent in the Department of Otolaryngology-Head and Neck Surgery and the Radiation Oncology Center of the Washington University School of Medicine (St. Louis, MO) between January 1971 and December 1991 and developed delayed regional metastases (2 y after treatment), distant metastases, and second primary malignancies. RESULTS: In 2550 patients, the mean age (59.8 y), sex (8.5 male patients and 1 female patient), and tumor differentiation did not affect the incidence of delayed distant, regional, or second primary malignancies. The overall incidence of delayed regional metastases was 12.4% (317/2550 patients); distant metastases, 8.5% (217/2550); and second primary tumors, 8.9% (228/2550), with a 5-year disease-specific survival of 41%, 6.4%, and 35%, respectively. Second primary malignancies were not statistically related to the origin of the primary tumor, tumor staging, or delayed regional and distant metastases (P =.98). Delayed regional metastases and distant metastases were related to advanced primary disease (T4 stage), lymph node metastases (node positive [N+]), tumor location (hypopharynx), and locoregional tumor recurrence (P < or =.028). Advanced regional metastases at initial diagnosis (N2 and N3 disease) increased the incidence of delayed and distant metastases threefold (P =.017). These two metastatic parameters were significantly greater in hypopharyngeal tumors than in laryngeal tumors (P =.037). The incidences of delayed regional metastases by anatomical location of the primary tumor were as follows: glottic, 4.4%; supraglottic, 16%; subglottic, 11.5%; aryepiglottic fold, 21.9%; pyriform sinus, 31.1%; and posterior hypopharyngeal wall, 18.5%. The incidences of distant metastases were as follows: glottic, 4%; supraglottic, 3.7%; subglottic, 14%; aryepiglottic fold, 16%; pyriform fossa, 17.2%; and posterior hypopharyngeal wall, 17.6%. Seventeen hypopharyngeal tumors (2%) presented with M1 disease. Delayed regional metastases to the ipsilateral treated neck had a significantly worse survival prognosis than delayed metastases to the contralateral nontreated neck (P =.001). CONCLUSIONS: Conclusions are as follows: 1) The incidence of second primary tumors is independent from the primary tumor staging and distant and delayed regional metastases. The highest incidence occurred in patient groups with the highest disease-free survival rates (P =.0378). 2) Highest incidence of delayed and distant metastases occurred in hypopharyngeal tumors and was three times greater than in laryngeal cancers (P =.028). 3) Salvage therapeutic rates were poor for delayed metastases to the ipsilateral treated nodes and distant metastases as compared with contralateral neck metastases and second primary tumors (P =.001). 4) Delayed and distant lymph node metastases were significantly higher in advanced primary disease (T4 stage), locoregional recurrences, and regional disease (N2 and N3) (P =.028) in both the larynx and hypopharynx. 5) The higher incidence of delayed and distant metastatic disease was related to more advanced initial tumor presentation in hypopharyngeal cancer as compared with laryngeal cancer (P =.039). 6) Incidence of distant metastases was greatest between 1.5 and 6 years after initial treatment with a mean incidence being less than or equal to 3.2 years. PMID- 11404626 TI - Is dissection of level IV necessary in patients with T1-T3 N0 tongue cancer? AB - OBJECTIVE: Dissection of the lower jugular level of lymph nodes (level IV), as part of an elective neck dissection, has been advocated recently for all patients with oral tongue cancer because of the possibility of "skip metastases" to levels III and IV. The current study was undertaken to evaluate the need to perform a dissection of level IV in patients with oral tongue cancer with no clinical evidence of nodal metastases. METHODS: Fifty-one patients with T1-3, N0 squamous cell carcinoma of the oral tongue were treated with a partial glossectomy and a selective neck dissection of levels I, II, and III. When enlarged nodes were encountered during surgery in level II or III, the dissection was extended to include the nodes in level IV. Involvement of level IV was determined either by the presence of carcinoma on pathological examination or by the development of recurrence in the untreated level IV during a follow-up period of at least 2 years. RESULTS: Level IV was resected as part of the specimen in 17 of the 51 patients and metastatic tumor was found in this level in only one patient. At an average follow-up of 4.1 years, only one patient recurred at level IV, which had been addressed at the initial neck dissection. Consequently, the rate of metastases to undissected level IV was 2%. CONCLUSIONS: Metastases to level IV lymph nodes is rare in patients with T1-T3, N0 oral tongue cancer. Dissection of these nodes only when there is intraoperative suspicion of metastases in levels II or III does not increase the risk or recurrence of tumor in the neck. PMID- 11404627 TI - Photofrin-mediated photodynamic therapy for treatment of aggressive head and neck nonmelanomatous skin tumors in elderly patients. AB - OBJECTIVES/HYPOTHESIS: Aggressive nonmelanomatous skin tumors (basal cell carcinoma, squamous cell carcinoma, and Bowen's disease) of the head and neck often occur in Caucasian elderly patients because of prior history of radiation therapy for teenage acne and adenoid hypertrophy; severe solar-induced skin damage, basal cell nevus syndrome, and other genetic skin diseases; chemical carcinogen exposure; and drug-induced immunosuppression. In patients with large, multifocal recurrent tumors, standard therapy with acceptable cosmetic outcomes may be difficult. Photodynamic therapy (PDT) with photosensitizing agents selectively taken up by skin provides a primary or adjunct intraoperative option for treatment of this special group of cancer patients. STUDY DESIGN: Retrospective review. METHODS: Patients (age range, 60-92 y) were injected with 1.0 mg/kg PHOTOFRIN (dihematoporphyrin derivative) followed 60 hours later by intraoperative laser light activation. Light was delivered through microlens fiber by means of an argon dye laser at 630 nm at a light dose of 100 to 300 J/cm2 microlens delivery for PDT alone and 50 to 100 J/cm2 microlens delivery for tumor bed resection sites in the case of adjunct PDT combined with surgical resection. RESULTS: Twelve cases of aggressive recurrent nonmelanomatous cutaneous tumors of the head and neck were treated. Five patients received intraoperative PDT combined with surgical resection, including radical mastoidectomy, lateral temporal bone resection, partial maxillectomy with temporalis myofacial flap reconstruction, and wide local resection with secondary intention healing of exposed scalp wounds. Seven patients were treated with PDT alone for extensive multiple cutaneous lesions or wide-field primary or recurrent nonmelanomatous tumors. Ten patients achieved complete responses (follow-up, 6-60 mo) with excellent wound healing and cosmetic outcomes. CONCLUSIONS: PHOTOFRIN mediated PDT is an excellent locoregional oncological modality for aggressive primary or recurrent basal cell carcinoma and squamous cell carcinoma, particularly in elderly patients who were previously treated with extensive Mohs microsurgery, surgical resection, and external-beam radiation therapy. Multiple repeat treatments are well tolerated, painless, without systemic morbidity, and amenable to local anesthesia or intravenous sedation for PDT alone, and wound healing and cosmetic outcomes are excellent. PMID- 11404628 TI - Significance of retropharyngeal node dissection at radical surgery for carcinoma of the hypopharynx and cervical esophagus. AB - OBJECTIVES: To clarify the efficacy of dissection of retropharyngeal lymph nodes (RPLNs) in the surgical treatment of carcinoma of the hypopharynx and cervical esophagus. STUDY DESIGN: We started planned dissection of the RPLN during initial radical surgery in patients with squamous cell carcinoma of the hypopharynx or the cervical esophagus in 1988. Until 1997, we performed this procedure as a standard operation in 82 consecutive patients. METHODS: Mortality resulting from RPLN metastasis was compared between 82 patients who underwent RPLN dissection and 69 patients who did not undergo the procedure. RESULTS: Of 82 patients, 16 patients (20%) had positive RPLNs. These patients were at high risk of recurrence unless the node(s) were dissected. Although RPLN dissection did not improve the cumulative 5-year survival rate, it significantly decreased the number of patients who died of RPLN metastasis (chi2 = 3.68, P <.1). Four of the 16 patients who had positive RPLNs survived without any recurrence. CONCLUSION: Bilateral dissection of the RPLN during initial surgery is highly recommended in every surgical case of carcinoma of the hypopharynx and cervical esophagus. PMID- 11404629 TI - Human papilloma virus and p53 expression in carcinomas associated with sinonasal papillomas: a Danish Epidemiological study 1980-1998. AB - OBJECTIVES: To determine a putative role and relation between human papilloma virus (HPV) and p53 in the etiology of sinonasal carcinomas associated with papillomas. STUDY DESIGN: The study group consists of all patients with sinonasal carcinomas associated with papillomas diagnosed in Denmark from 1980 to 1998. After reviewing our national pathological files, tumor tissues from 36 patients were collected, comprising 15% of the total cases of sinonasal carcinomas. In 35 cases a squamous cell carcinoma was demonstrated and in one case an adenocarcinoma was evident. Inverted papilloma was associated with carcinoma in 31 cases and exophytic papillomas in 5 cases. The material was investigated for HPV using polymerase chain reaction analyses with two sets of consensus primers (GP5+/GP6+ and MY09/MY11). The HPV-positive cases were submitted to dot-blot hybridization to establish the HPV type. Using immunohistochemistry, the p53 expression was determined. A p53 overexpression is defined as positive staining in 10% or more of the tumor cells. RESULTS: Among 30 examined cases of carcinomas associated with inverted papillomas, 4 cases were HPV-positive (13%). P53 overexpression was not shown among the HPV-positive cases, whereas p53 overexpression was seen in 21 of the 24 (88%) examined HPV-negative cases. Among the 5 carcinomas associated with exophytic papillomas, HPV was demonstrated together with p53 overexpression in 3 cases (60%). In addition, one case more was with p53 overexpression. CONCLUSION: An inverse relation between HPV and p53 overexpression in sinonasal carcinomas associated with inverted papillomas appears to have been demonstrated. HPV and p53 might also have an etiological role among the carcinomas associated with exophytic papillomas. PMID- 11404630 TI - A simplified method of micro-injecting vasoconstricting solution into the vocal cord. PMID- 11404631 TI - Modifications for silastic sheet use in nasal surgery. PMID- 11404632 TI - Laryngeal reinnervation with the hypoglossal nerve. PMID- 11404633 TI - Hearing results of intratympanic steroid treatment of endolymphatic hydrops. PMID- 11404634 TI - Management of communication disorders in the 21st century. PMID- 11404637 TI - Pushing the envelope--health care, not welfare. PMID- 11404638 TI - Determinants of public and private insurance enrollment among Medicaid-eligible children. AB - BACKGROUND: Many Medicaid-eligible children are not enrolled in Medicaid and are not covered by private insurance. Reducing persistent lack of insurance for children requires a better understanding of why Medicaid-eligible children do not participate. RESEARCH QUESTIONS: Does the availability of free or low-cost medical services substitute for Medicaid or private insurance enrollment among Medicaid-eligible children? Does the availability and affordability of insurance coverage, particularly the offer of employer-sponsored insurance (ESI) and the presence of managed care, affect child insurance coverage? RESEARCH DESIGN: We use data from the National Health Interview Survey for 1994 and 1995, supplemented with county level measures of insurance and provider supply, to estimate a multinomial choice model of insurance coverage among children identified as Medicaid-eligible. We focus on county supply of public hospitals and community/migrant health centers (C/MHC); and the availability and cost of ESI. We control for child and parent characteristics. RESULTS: A positive effect of C/MHC supply is found on Medicaid enrollment, but no evidence is found of substitution between low-cost providers and Medicaid or private coverage. Local availability of ESI and private HMO penetration increased private insurance enrollment. CONCLUSIONS: Local community providers can play an important role in outreach and enrollment for Medicaid. Availability and cost of ESI constrain private coverage for Medicaid-eligible children. Policies that encourage offers of insurance coverage by employers, decrease premiums, and encourage adoption of managed care could have important positive effects on coverage for this population. PMID- 11404639 TI - Physician incentives and the timing of cesarean sections: evidence from California. AB - OBJECTIVES: The timing of cesarean sections is studied to examine how physician convenience and financial incentives play a role in the decision to perform a cesarean section. METHODS: Using birth certificate and hospital financial data from California, the likelihood of cesarean sections being performed at particular times of day was examined, controlling for maternal characteristics and the mother's insurance coverage. Two diagnoses associated with cesarean sections are examined separately: fetal distress and prolonged/dysfunctional labor. The hypotheses are that cesarean sections performed for physician convenience are more likely to occur in the evening hours and that type of insurance will affect the incentive to perform cesarean sections to obtain leisure. RESULTS: The probability of cesarean sections for patients insured by a group-model HMO is more stable during the course of a day than that for patients insured by all other insurance plans. Group-model HMO patients with previous cesarean sections are less likely to have cesarean sections in the evening hours and are less likely to be diagnosed with fetal distress or prolonged/dysfunctional labor. CONCLUSIONS: The differences in cesarean sections and diagnosis rates between group-model HMO patients and other patients could arise from several mechanisms: group-model HMOs provide consistent financial incentives to their staff, they may be better able to guide physician practice, and they might provide staff support to physicians so there is less leisure-based incentive to perform cesarean sections. In contrast, nongroup-model HMOs do not appear to reduce the incentive of physicians to maximize leisure relative to traditional insurance. PMID- 11404640 TI - Ambulatory care sensitive hospitalizations and emergency visits: experiences of Medicaid patients using federally qualified health centers. AB - BACKGROUND: Federally Qualified Health Centers (FQHCs) serve as regular sources of preventive and primary care for low-income families within their communities and are key parts of the health care safety net. OBJECTIVES: Compare admissions and emergency room visits for ambulatory care sensitive conditions (ACSCs) among Medicaid beneficiaries relying on FQHCs to other Medicaid beneficiaries. RESEARCH DESIGN: Retrospective analysis of 1992 Medicaid claims data for 48,738 Medicaid beneficiaries in 24 service areas across five states. SUBJECTS: Medicaid beneficiaries receiving more than 50% of their preventive and primary care services from FQHCs are compared with Medicaid beneficiaries receiving outpatient care from other providers in the same areas. Exclusions-managed care enrollees, beneficiaries more than age 65, dual eligibles (Medicaid and Medicare), and institutionalized populations. MEASURES: Admissions and emergency room (ER) visits for a set of chronic and acute conditions, known in the literature as ambulatory care sensitive conditions (ACSCs). RESULTS: Medicaid beneficiaries receiving outpatient care from FQHCs were less likely to be hospitalized (1.5% vs. 1.9%, P < 0.007) or seek ER care (14.9% vs. 15.7%, P < 0.02) for ACSCs than the comparison group. Controlling for case mix and other demographic variables, the odds ratios were, for hospitalizations, OR, 0.80; 95% CI, 0.67 to 0.95; P < 0.01, and for ER visits, OR, 0.87; 95% CI, 0.82 to 0.92; P < 0.001. CONCLUSIONS: Having a regular source of care such as FQHCs can significantly reduce the likelihood of hospitalizations and ER visits for ACSCs. If the reported differentials in ACSC admissions and ER visits were consistently achieved for all Medicaid beneficiaries, substantial savings might be realized. PMID- 11404641 TI - Can the health utilities index measure change? AB - BACKGROUND: The Health Utilities Index (HUI) is a multidimensional, preference weighted measure of health status. It comprises eight health attributes, aggregated into a single utility score. OBJECTIVES: The purpose of the study was to investigate the ability of the HUI to detect changes in health status in a general population cohort. RESEARCH DESIGN: Health status changes were analyzed in the full cohort and in persons who were diagnosed with chronic conditions, hospitalized, or became restricted in daily activities. SUBJECTS: To assess responsiveness, longitudinal data was used from the National Population Health Survey conducted in Canada in 1994 - 1995 and 1996 - 1997. We used cross sectional data from the 1996 sample to classify chronic conditions into mild, moderate, and severe. MEASURES: Two measures of responsiveness were calculated: Standardized Response Mean (SRM) and Sensitivity Coefficient (SC). The HUI was compared with a global health index-the Self-Rated Health (SRH) scale. RESULTS: HUI scores improved between the two NPHS cycles in all age-sex groups, except men 65 years of age and older. Among the respondents who remained free of chronic conditions, improvements were seen primarily in the cognitive and emotional domains. The HUI deteriorated among persons who were diagnosed between the two cycles with a severe chronic condition, were hospitalized, or became restricted in activity, but not in those diagnosed with a moderate condition. The SRMs were generally smaller for the HUI compared with the SRH. CONCLUSIONS: The HUI responds to changes in health status associated with serious chronic illnesses. However, changes in the HUI do not always coincide with changes in self-reported health. Properties of the HUI scales require further study. PMID- 11404642 TI - Factors influencing quality of life in older women with heart disease. AB - BACKGROUND: Better understanding of factors influencing the quality of life (QOL) of cardiac patients can guide treatment decisions. OBJECTIVES: To describe the impact of clinical and psychosocial factors on the QOL of older women with heart disease. RESEARCH DESIGN: Baseline and 12-month data from women participating in an intervention study. SUBJECTS: Eligible participants, identified from medical records, were female, > or = 60 years of age, and diagnosed with cardiac disease. A volunteer sample of 570 women (87% white) completed baseline interviews, with 485 women completing the 12-month assessment. MEASURES: Utilizing Wilson and Cleary's conceptual framework (1995), measures of clinical, psychosocial, and functional status were examined for their associations with QOL. RESULTS: At baseline, General Health Perceptions and Symptom Status accounted for 38% and 26%, respectively, of the variation in the QOL rating. Using logistic regression models, seven measures were significant predictors (P < 0.05) of maintenance/improvement versus decline in QOL over 12 months: baseline QOL rating; baseline value and change in satisfaction with social activities over 12 months; change in satisfaction with physical activities; change in satisfaction with mental activities; and baseline value and change in perceived stress. For women who maintained or improved their satisfaction with social activities, the odds for also maintaining or improving QOL were 4.5 times the odds for women whose satisfaction with social activities deteriorated. CONCLUSIONS: Satisfaction with social activities and perceived stress are important predictors of subsequent QOL. Consideration of the impact of treatments on these factors may help to prevent deterioration of QOL among older female cardiac patients. PMID- 11404643 TI - Medical care expenditures for hypertension, its complications, and its comorbidities. AB - OBJECTIVES: Medical expenditures attributed to hypertension were estimated, including expenditures for cardiovascular complications, other conditions for which hypertensives are at higher risk, and comorbidities (secondary diagnoses) that raise the cost of medical care. This article presents total, per capita, and per condition US expenditures in 1998 according to sex, age, and type of health service. METHODS: A variety of national data sources were used to disaggregate national health expenditures in 1998 by diagnosis. Expenditures for cardiovascular complications and other conditions for which hypertensives had higher rates of utilization were determined by analysis of attributable risks. Additional expenditures generated by extra hospital inpatient days and higher charges for nursing home and home health care for comorbidities were estimated by regression analyses. RESULTS: In 1998, $108.8 billion in health care spending was attributed to hypertension, 12.6% of total national spending that could be allocated to diagnoses, including $22.8 billion for hypertension, $29.7 billion for cardiovascular complications, and $56.4 billion for other diagnoses. Per capita expenditures increased with age from $249 for those younger than 65 years to $3,007 for those 85 years and older. The average amount spent per hypertensive condition was $3,787. Expenditures were generally higher for females. CONCLUSIONS: The economic burden of hypertension is large, but health services directly related to hypertension account for only a fraction of attributed expenditures. Comprehensive accounting of expenditures more accurately assesses the cost of hypertension and potential savings from prevention and treatment. Alteration of lifestyles and medical intervention provide opportunities to reduce national health expenditures. PMID- 11404644 TI - Preventive services among Medicare beneficiaries with supplemental coverage versus HMO enrollees, medicaid recipients, and elders with no additional coverage. AB - BACKGROUND: Studies conducted when Medicare began to cover preventive services, found that beneficiaries with supplemental insurance were much more likely to have such services than those without additional coverage. OBJECTIVE: To examine preventive services among Medicare beneficiaries with supplemental insurance, Medicaid, health maintenance organization (HMO) enrollees, and those without additional insurance. RESEARCH DESIGN: Analysis of the 1996 Medical Expenditure Panel Survey, a nationally representative multistage survey. SUBJECTS: 2,251 persons aged 65 and older with Medicare coverage. MEASURES: Self-reported preventive services, specifically, blood pressure measurement, cholesterol testing, influenza vaccination, mammography, Papanicolau (Pap) testing, and breast and prostate examinations. Multivariate modeling was used to adjust for age, education, race/ethnicity, and functional status. RESULTS: Elders without additional coverage were approximately 10% points less likely to have influenza vaccination, cholesterol testing, mammography, or Pap smears than those with supplemental coverage (P < 0.05). Multivariate adjustment attenuated some of these differences with age and education being the most important predictors of having preventive services. HMO enrollees were more likely to have mammograms than those with supplemental coverage (P < 0.05). CONCLUSIONS: Several years after Medicare extended coverage to include preventive services, differences in utilization of such services among elders with and without supplemental insurance have narrowed substantially. PMID- 11404645 TI - Multisite randomized controlled trials in health services research: scientific challenges and operational issues. AB - Although well-designed randomized controlled trials (RCT) provide the strongest evidence regarding causation, only relatively recently have they been used by health services researchers to study the organization, delivery, quality, and outcomes of care. More recent yet is the extension of multisite RCTs to health services research. Such studies offer numerous methodological advantages over single-site trials: (1) enhanced external validity; (2) greater statistical power when studying conditions with a low incidence or prevalence, small event rate in the outcome (eg, mortality), and/or large variance in the outcome (eg, health care costs); and (3) rapid recruitment to provide health care organizations and policy makers with timely results. This paper begins by outlining the advantages of multisite RCTs over single-site trials. It then discusses both scientific challenges (ie, standardizing eligibility criteria, defining and standardizing the intervention, defining usual care, standardizing the data collection protocol, blinded outcome assessment, data management and analysis, measuring health care costs) and operational issues (ie, site selection, randomization procedures, patient accrual, maintaining enthusiasm, oversight) posed by multisite RCTs in health services research. Recommendations are offered to health services researchers interested in conducting such studies. PMID- 11404646 TI - Adjusting for patient characteristics when analyzing reports from patients about hospital care. AB - OBJECTIVES: To determine which patient characteristics are associated with reports and ratings of hospital care, and to evaluate how adjusting reports and ratings for hospital differences in such variables affects comparisons among hospitals. DESIGN: A telephone survey of a sample of patients hospitalized in 22 hospitals in a single city and a statewide mail survey of hospitalized patients. MEASURES: The surveys assessed: respect for patients' preferences, coordination of care, information exchange between patient and providers, physical care, emotional support, involvement of family and friends, and transition and continuity. The surveys also asked patients to rate their doctors, nurses, and other hospital staff. RESULTS: The variables with the strongest and most consistent associations with patient-reported problems were age and reported health status. Patient gender and education level also sometimes predicted reports and/or ratings. Models including these variables explained only between 3% and 8% of the variation in reports and ratings. CONCLUSIONS: The impact of adjusting for patient characteristics on hospital rankings was small, although a larger impact would be expected when comparing hospitals with more variability in types of patients. Nevertheless, we recommend adjusting at least for the most important predictors, such as age and health status. Such adjustment helps alleviate concerns about bias. It also may be useful to present data for certain groups of patients (ie, medical, surgical, obstetric) separately to facilitate interpretation and quality improvement efforts. PMID- 11404647 TI - The human gene map for performance and health-related fitness phenotypes. AB - The aim of this paper is to describe the first human gene map for physical performance and health-related fitness traits based on the papers published until the end of 2000. Studies of candidate genes using case-control and other designs are reviewed. Quantitative trait loci from the limited evidence reported to date in genomic scans are also incorporated. Performance and fitness phenotypes in the sedentary state as well as their changes during exercise, if applicable, or in response to exercise training are considered. Physical performance traits include cardiorespiratory endurance indicators and muscular strength or muscular performance variables. Health-related fitness phenotypes are grouped under the following categories: hemodynamic traits; anthropometry and body composition; insulin and glucose metabolism; and lipids, lipoproteins, and hemostatic factors. A yearly update of this human gene map will be published. PMID- 11404648 TI - The human gene map for performance and health-related fitness phenotypes. PMID- 11404649 TI - Effects of resistance exercise and creatine supplementation on myasthenia gravis: a case study. AB - PURPOSE: The purpose of this case study was to determine the effects of 15 wk of resistance exercise and creatine (Cr) supplementation on body composition, training volume, peak strength, and complete blood chemistry in a patient with myasthenia gravis (MG). METHODS: The patient was a 26-yr-old man who was taking prednisone and azathioprine for his condition. The patient self-administered 5 g of Cr per day in addition to resistance exercise 3 times per week. Fasting blood samples were obtained and body weight (BW) and fat free mass (FFM; via hydrostatic weighing) were measured before and after training and Cr supplementation. In addition, isokinetic (Cybex II) peak strength for leg extension (LE), leg flexion (LF), and volume load (repetition x mass lifted) for the first and last resistance training session were determined. RESULTS: After Cr supplementation and training, the results demonstrated increases in BW (6.8%), FFM (4.3%), upper body volume load (37.0%), lower body volume load (15.0%), and peak strength for LE (37.0%) and LF (12.5%). Moreover, blood chemistry values remained within normal limits for the duration of the 15-wk study. CONCLUSION: These data suggest that resistance exercise plus Cr supplementation may promote gains in strength and FFM in patients with MG. PMID- 11404650 TI - Impact of exercise on bone health and contraindication of oral contraceptive use in young women. AB - PURPOSE: The effect of quantified resistance and high impact exercise training on bone mass as modified by age and oral contraceptive (OCont) use in young women was studied. METHODS: Women were categorized by age (18-23 vs 24-31 yr) and OCont use, and were then randomized into either three sessions of resistance exercise plus 60 min.wk-1 of jumping rope or a control group for 24 months. Total body, spine, femoral neck, greater trochanter, Ward's area, and radial bone mineral density (BMD) and/or content (BMC), biochemical markers of bone turnover, dietary intake of calcium, lean body mass, maximal oxygen uptake, and strength were determined at baseline and every 6 months. RESULTS: Total body (TB) BMC percent change from baseline was higher in exercisers compared with nonexercisers at 6 and 24 months. OCont users had lower bone turnover at baseline and a decrease in TBBMC from baseline compared with non-OCont users at 24 months. Spine BMC and BMD decreased in the exercise and OCont group at 6 months and remained significantly below nonexercisers who used oral contraceptives at 2 yr. Femoral neck BMD also decreased in the exercise and oral contraceptive group at 6 months. CONCLUSIONS: Exercise prevented a decline in TBBMC seen in the nonexercisers. On the other hand, exercise in oral contraceptive users prevented the increase observed in the spine of the nonexercise plus OCont group. PMID- 11404651 TI - Acute resistive exercise does not affect ambulatory blood pressure in young men and women. AB - PURPOSE: Resistive exercise elicits a pressor response that results in a dramatic increase in blood pressure (BP) during the exercise. However, it is not known if the BP elevation persists after resistive exercise. METHODS: This study examined the effects of an acute resistive exercise session on 24-h ambulatory BP in sedentary (5 men, 6 women), resistance-trained (6 men, 6 women), and endurance trained (4 men, 6 women) young subjects (age 22 +/- 3.2 yr) with normal BP. Two 24-h ambulatory BP recordings were made on each subject, one after two sets of resistive exercise on 12 weight machines and one after 48 h without prior exercise. RESULTS: Systolic, diastolic, and mean arterial BP and heart rate (HR) were not different in the hours after and for up to 24 h after the single resistive exercise session compared with the control day. There also was no difference in the ambulatory BP or HR response after the single session of resistive exercise based on the training status of the subjects. CONCLUSION: Thus, the elevated BP that occurs during resistive exercise does not persist in the 24 h after acute resistive exercise in sedentary, resistance-trained, or endurance-trained, young, normotensive men and women. PMID- 11404652 TI - Clotting and fibrinolytic activity change during the 1 h after a submaximal run. AB - PURPOSE: To determine the changes in clotting and fibrinolytic activity during the 1-h period after an acute submaximal exercise at a specific relative exercise intensity to ascertain whether during this time there is a greater risk for developing a clot formation or thrombus. METHODS: Ten healthy men reported between 0700 and 1000 h and ran at 70-75% VO2max or walked at 1.2 mph for 30 min in a random counter-balanced order. Venous blood was obtained at rest, immediately after, and every 20 min during the 1-h recovery. RESULTS: There were no differences in the resting parameters for each treatment. Walking did not alter the activity of any of the measures analyzed compared with rest. Clotting indicators activated partial thromboplastin time (APTT) was significantly decreased by approximately 2 s and remained at this level during the 1-h recovery, and factor VIII activity was elevated 66% immediately after the run and remained elevated at this level during the 1-h recovery period. Fibrinolytic indicators, t-PA, and D-dimers were significantly increased immediately after the run. However, t-PA demonstrated a quadratic negative slope during the 1-h recovery time. D-dimers remained elevated during the 1-h recovery time. CONCLUSIONS: These results suggest that running at 70-75% VO2max resulted in elevated clotting and fibrinolytic activity. However, the clotting activity was sustained during a time when fibrinolytic activity declined, which suggests a more favorable situation for clot formation during this time after exercise. PMID- 11404653 TI - Laboratory protocol for exercise asthma to evaluate salbutamol given by two devices. AB - PURPOSE: As new delivery devices and formulations are being introduced for drugs given by inhalation, there is a need to evaluate their equivalence with old preparations. One way to do this is to investigate their equivalence in protecting from exercise-induced asthma (EIA). METHODS: We used a protocol for EIA to compare the protective effect of salbutamol delivered by the pressurised metered dose inhaler (pMDI) and the new Diskus dry powder device. Twenty-seven asthmatic subjects with moderately severe EIA completed an exercise test on four separate days at two study centers. Exercise was performed by cycling for 8 min while inhaling dry air (0% RH, 20-24 degrees C). The target workload in W was predicted as (53.76 x predicted FEV1) - 11.07 and 95% of this target was achieved at 4 min of exercise. This target was chosen in order to achieve ventilation between 50 and 60% of predicted maximum in the last 4 min. RESULTS: There was no significant difference in the workload, ventilation, or heart rate achieved on the study days. The severity of EIA was measured as the % fall in FEV1. EIA severity was similar on the placebo and control day and the coefficient of variation was 19.4%. The mean +/- SD % fall on the control, placebo, salbutamol by Diskus, and pMDI were 42.0% +/- 15, 39.4% +/-17.6, 13.4% +/- 13.2, and 8.5% +/ 13.8, respectively. Salbutamol significantly inhibited the % fall in FEV1 after exercise, and there was no difference between the preparations. CONCLUSION: The protocol described here is suitable for evaluating equivalence of salbutamol preparations in protecting against EIA and could be used to evaluate the protective effect of other medications. PMID- 11404654 TI - Effects of age on muscle energy metabolism and oxygenation in the forearm muscles. AB - PURPOSE: The effects of aging on muscle metabolism and oxygenation have not yet been elucidated. We evaluated the effects of aging on energy metabolism and oxygenation in sedentary healthy subjects by simultaneously measuring 31P magnetic resonance spectroscopy (MRS) and near-infrared spectroscopy (NIRS). METHODS: Nine young (28.1 +/- 5.0 yr) and nine older (61.4 +/- 4.6 yr) healthy subjects were studied. The 31P-MR spectrum was obtained every 15 s during and after hand gripping exercise. Intracellular pH (pHi) and PCr/(PCr+Pi) [PCr: phosphocreatine, Pi: inorganic phosphate] were calculated as an index of energy metabolism. The time constant of the PCr/(PCr+Pi) recovery (tau PCr) was calculated. With NIRS, we evaluated the recovery rates of oxygenated (RHbO2) and deoxygenated hemoglobin (RHb) during the initial 10 s of recovery. RESULTS: The PCr/(PCr+Pi) and pHi at rest and at completion of the exercise and tau PCr did not differ between young and older subjects. However, RHbO2 and RHb were significantly slower in older subjects than in young subjects. CONCLUSIONS: The results suggest that muscle energy metabolism in the forearm muscle was not affected by aging. The slower RHbO2 and RHb in older subjects suggested impaired O2 supply, which was probably due to impaired peripheral circulation caused by the process of aging. PMID- 11404655 TI - Intestinal fluid absorption during exercise: role of sport drink osmolality and [Na+]. AB - The purpose of this study was to evaluate the effects of modifying the osmolality and [Na+] of orally ingested rehydration beverages during exercise on intestinal absorption in the duodenum and upper jejunum. Six subjects randomly ingested (23 mL.kg-1 BW) the following 6% carbohydrate solutions with and without Na+ during 85-min of cycle exercise (65% VO2 peak) in a cool (22 degrees C, 40% RH) environment: a) 0 Na+, 245 mOsm.kg-1; b) 20 mEq Na+, 283 mOsm.kg-1; c) 20 mEq Na+, 169 mOsm.kg-1; d) 50 mEq Na+, 275 mOsm.kg-1; and e) 50 mEq Na+, 176 mOsm.kg 1. To alter solution osmolality and maintain carbohydrate concentration constant, glucose, sucrose, fructose, and maltodextrin were used in different combinations. Nasogastric and multilumen tubes were fluoroscopically placed in the stomach and intestine, respectively, to simultaneously determine gastric emptying and intestinal absorption as previously described (Lambert et al., Int. J. Sports Med.17:48, 1996). Gastric emptying was not different among solutions and averaged 13 +/- 0.5 mL.min-1. Net fluid absorption was not different among beverages nor between duodenum and jejunum (x = 10.8 +/- 1.6 and 7.9 +/- 1.1 mL.cm-1.h-1, respectively). Mean osmolality increased significantly (P < 0.05) from the duodenum to the jejunum (242 +/- 6 and 293 +/- 7 mOsm.kg-1, respectively) but did not differ among solutions. These data provide evidence that a hypotonic 6% carbohydrate beverage with 50 mEq.L-1 Na+ did not enhance intestinal fluid absorption or attenuate the decline in plasma volume during exercise more than an isotonic carbohydrate-electrolyte solution or a hypotonic carbohydrate solution without sodium. PMID- 11404656 TI - Effects of oxygen on lower limb blood flow and O2 uptake during exercise in COPD. AB - PURPOSE: To quantify the effects of acute oxygen supplementation on lower limb blood flow (QLEG), O2 delivery (QO2LEG), and O2 uptake (VO2LEG) during exercise and to determine whether the metabolic capacity of the lower limb is exhausted at peak exercise during room air breathing in patients with COPD. METHODS: Oxygen (FIO2 = 0.75) and air were randomly administered to 14 patients with COPD (FEV1: 35 +/- 2% pred, mean +/- SEM) during two symptom-limited incremental cycle exercise tests. Before exercise, a cannula was installed in a radial artery and a thermodilution catheter inserted in the right femoral vein. At each exercise step, five-breath averages of respiratory rate, tidal volume, and ventilation (VE), dyspnea and leg fatigue scores, arterial and venous blood gases, and QLEG were obtained. From these measurements, VO2LEG was calculated. RESULTS: Peak exercise capacity increased from 46 +/- 3 W in room air to 59 +/- 5 W when supplemental oxygen was used (P < 0.001). QLEG, QO2LEG, and VO2LEG were greater at peak exercise with O2 than with air (P < 0.05). During submaximal exercise, dyspnea score and VE were significantly reduced with O2 (P < 0.05), whereas QLEG, VO2LEG, and leg fatigue were similar under both experimental conditions. The improvement in peak exercise work rate correlated with the increase in peak QO2LEG (r = 0.66, P < 0.01), peak VO2LEG (r = 0.53, P < 0.05), and reduction in dyspnea at iso-exercise intensity (r = 0.56, P < 0.05). CONCLUSION: The improvement in peak exercise capacity with oxygen supplementation could be explained by the reduction in dyspnea at submaximal exercise and the increases in QO2LEG and VO2LEG, which enabled the exercising muscles to perform more external work. These data indicate that the metabolic capacity of the lower limb muscles was not exhausted at peak exercise during room air breathing in these patients with COPD. PMID- 11404657 TI - Leg stiffness and mechanical energetic processes during jumping on a sprung surface. AB - PURPOSE: The purposes of this study were: a) to examine the effect of verbal instructions given to the subjects on the control of lower extremity stiffness and b) to determine the effect of leg stiffness on mechanical energetic processes during drop jumps on a sprung surface. METHODS: A total of 10 female athletes performed a series of drop jumps on a sprung surface from heights of 20 and 40 cm. The instructions given to the subjects were a) "jump as high as you can" and b) "jump high a little faster than at your previous jump." The jumps were performed at each height until the athlete could not achieve a shorter ground contact time. Four jumps per subject per height were analyzed. The ground reaction forces were measured using a "Kistler" force plate (1000 Hz). The athletes' body positions were recorded using a high-speed (250 Hz) video camera. The deformation of the sprung surface was determined by another high-speed camera operating at 500 Hz. Surface EMG was used to measure muscle activity in five leg muscles. RESULTS: The contact time showed high correlation with leg stiffness as well as with ankle and knee stiffness. The change in leg stiffness was not due to the duration of the preactivation but rather to the level of activation during this phase. An increase in leg stiffness caused an increase in the energy stored and recovered in and by the sprung surface and a decrease of the energy produced by the subjects. CONCLUSIONS: By influencing contact time through verbal instructions, it is possible to control leg stiffness. Maximal vertical take-off velocity of the center of mass and maximal take-off body energy can be achieved having different levels of leg stiffness. The maximization of mechanical power is achieved by optimal leg stiffness values and leg muscle preactivation levels. PMID- 11404658 TI - Postexercise oxygen consumption and substrate use after resistance exercise in women. AB - OBJECTIVE: This study investigated the acute effects of 45 min of resistance exercise (RE) on excess postexercise oxygen consumption (EPOC) and substrate oxidation 120 min after exercise in moderately trained women. METHODS: Ten RE trained women (age = 29 +/- 3 yr; ht = 168 +/- 8.3 cm; wt = 59 +/- 5.7 kg; VO2max = 38.3 +/- 4.7 mL.kg-1.min-1) underwent two trials: control sitting and RE. Subjects acted as their own controls in a random counterbalanced design. A 2-d nonexercise period was established between testing trials. Oxygen consumption (VO2) and respiratory exchange ratio (RER) were measured continuously by indirect calorimetry before, during, and after exercise and on a separate control day. RE consisted of 3 sets of 10 exercises at 10-repetition maximum with a 1-min rest period between each set. Fingertip samples of blood lactate concentration [BL] were collected immediately postexercise and every 30 min thereafter until [BL] returned to resting baseline values after exercise. RESULTS: The overall 2-h EPOC was 6.2-L (RE = 33.4 +/- 5.1 L vs control = 27.2 +/- 0.3 L), corresponding to an 18.6% elevation over the control period. RER was significantly (P < 0.01) below the control RER from minute 30 to minute 120 postexercise (RE = 0.75 +/- 0.01 vs control = 0.85 +/- 0.01). During the last 30 min of recovery, VO2 and [BL] had returned to control/baseline values and fat oxidation was significantly (P < 0.0001) higher (29.2 vs 16.3 kcal) after RE compared with the control trial. CONCLUSION: These data indicate that in young RE trained women, acute RE produces a modest increase in VO2 during a 2-h recovery period and an increase in fat oxidation. PMID- 11404659 TI - Estimating energy expenditure by heart-rate monitoring without individual calibration. AB - Heart rate monitoring has been shown to be a valid method for measuring free living energy expenditure at the group level, but its use in large-scale studies is limited by the need for an individual calibration of the relationship between heart rate and energy expenditure. PURPOSE: To determine whether energy expenditure can be estimated from heart rate monitoring without individual calibration in epidemiological studies. METHODS: Our previously validated heart rate monitoring method relies on measuring individual calibration parameters obtained from resting energy expenditure and the regression line between energy expenditure and heart rate during exercise. We developed prediction equations for these parameters using easily measured variables in a population-based study of 789 individuals. The predictive ability of these parameters was tested in a separate population-based sample (N = 97). RESULTS: Physical activity level (PAL = total energy expenditure/basal metabolic rate) using the four estimated parameters was correlated with PAL using the measured parameters (r = 0.82, P < 0.01). Comparison of measured and estimated PAL showed that 97.9% of the scores were placed in the same or adjacent quartile. CONCLUSION: A combination of simple measurements and heart rate monitoring produces estimates of energy expenditure that are highly correlated with those obtained using full individual calibration. This simplification of the heart rate monitoring method could extend its use in ranking individuals in epidemiological studies. PMID- 11404660 TI - Risk factors for training-related injuries among men and women in basic combat training. AB - PURPOSE: Past investigations indicate that training-related injuries are associated with certain performance-oriented measures of physical fitness and certain lifestyle characteristics. This study examined associations between injuries, direct (physiological) measures of physical fitness, and lifestyle characteristics. METHODS: Subjects were 756 men and 474 women performing the standardized activities involved in U.S. Army Basic Combat Training (BCT). Before BCT, a subsample of subjects (182 men and 168 women) were administered a series of tests that included a treadmill running test (peak VO2), dual-energy x-ray absorptiometry (for body composition), several measures of muscle strength, a hamstring flexibility test, and a vertical jump. A questionnaire addressed smoking habits and prior physical activity. All subjects were administered the Army Physical Fitness test consisting of push-ups, sit-ups, and a 3.2-km run. Gender, age, stature, and body mass were obtained from physical examination records. Injuries incurred during BCT were transcribed from medical records; for each medical visit, the diagnosis, anatomical location, disposition (final outcome of visit), and days of limited duty were recorded. RESULTS: Women had over twice the injury rate of men. For men and women, fewer push-ups, slower 3.2 km run times, lower peak VO2, and cigarette smoking were risk factors for time loss injury. Among the men only, lower levels of physical activity before BCT and both high and low levels of flexibility were also time-loss injury risk factors. Multivariate analysis revealed that lower peak VO2 and cigarette smoking were independent risk factors for time-loss injury. CONCLUSIONS: Lower aerobic capacity and cigarette smoking were independently associated with a higher likelihood of injury in both men and women during a standardized program of physical training. Further studies are needed to assess associations between injury and body composition and muscular strength. PMID- 11404661 TI - A comparison of the Yale Physical Activity Survey with other physical activity measures. AB - PURPOSE: The purpose of this study was to determine associations of the Yale Physical Activity Survey (YPAS) with the Stanford 7-d physical activity recall (PAR) and several physiologic measures, both cross-sectionally and longitudinally. METHODS: Participants were 59 individuals (45% African-American, 79% women) between the ages of 60 and 80 yr who were enrolled in a clinical trial. From the YPAS, total time in activity, estimated weekly energy expenditure, and indices of vigorous activity, leisurely walking, moving, standing, sitting, and an overall summary index were determined. From the PAR, daily energy expenditure, and hours spent in light, moderate, and hard/very hard activities were estimated. Physiologic measurements included estimated VO2max, resting pulse rate, and body mass index (BMI). RESULTS: Weekly energy expenditure, total time in activity, and the summary index correlated with daily energy expenditure at baseline (Spearman correlations = 0.37, 0.30, 0.33, respectively, Ps < 0.01) and time in moderate activity (r = 0.37, 0.29, 0.39, Ps < 0.01). The vigorous index was also significantly correlated with daily energy expenditure and time in light, moderate, and hard/very hard activity. The summary, moving, and standing indices correlated with VO2max (Ps < 0.05) and BMI (Ps < 0.05). Change in physical activity determined from the PAR correlated with change in most YPAS dimensions, although few correlations were statistically significant. CONCLUSIONS: The global activities and the vigorous activity index determined from the YPAS correlated with corresponding measures of the PAR. The validity of the YPAS for light-intensity activities is less clear, although optimal validation measures for light-intensity activity are yet to be defined. The ability of the YPAS to detect change in physical activity shows some promise, but data from this study are insufficient to be conclusive. PMID- 11404662 TI - An evaluation of three self-report physical activity instruments for older adults. AB - PURPOSE: To assess the known-groups and construct validity of measures from the CHAMPS Physical Activity Questionnaire, Physical Activity Survey for the Elderly (PASE), and the Yale Physical Activity Survey (YPAS). METHODS: The three questionnaires were administered to a convenience sample of older adults (N = 87) recruited from community centers and retirement homes. Validation measures included the SF-36 measures of physical functioning, general health, mental health, and pain; body mass index; performance-based tests of lower body functioning and endurance; and Mini-Logger activity monitor data from ankle and waist sensors. Validity was estimated by testing hypotheses about associations between physical activity and validation measures. RESULTS: As hypothesized, differences in activity levels on all measures were found between older adults in retirement homes (less active) and community centers (more active) (P-values < 0.0001). Correlations of physical activity measures with performance-based measures ranged from 0.44 to 0.68, conforming to hypotheses; hypotheses regarding associations with the SF-36 measures were also confirmed. Body mass index was not correlated with any of the physical activity measures, contrary to hypotheses. Correlations of physical activity measures with Mini-Logger counts ranged from 0.36 to 0.59 (ankle) and 0.42 to 0.61 (waist) as hypothesized. Correlations among the measures from the three instruments ranged from 0.58 to 0.68. CONCLUSIONS: The PASE, YPAS, and CHAMPS each demonstrated acceptable validity, as all measures met nearly all hypotheses. Higher validity coefficients were found for subgroups (men, 65-74 yr, retirement home), suggesting that these instruments may perform better for certain segments of the older adult population. PMID- 11404663 TI - Effects of eccentric exercise on trunk extensor torque and lumbar paraspinal EMG. AB - PURPOSE: Little is known about the effects of eccentric contractions on the function of the lumbar paraspinal muscles. The purpose of this study was to determine the effects of a single bout of eccentric contractions using the trunk extensor muscles on torque and lumbar paraspinal electromyographic (EMG) parameters. METHODS: Twenty healthy men between the ages of 18 and 49 yr participated in the study. Subjects performed a single bout of 50 maximal voluntary concentric (N = 10) or eccentric (N = 10) trunk extension movements while surface EMG signals were recorded from the multifidus and iliocostalis lumborum muscles. A series of isometric contractions were performed both before the exercise protocol and at five additional time points over the following 7 d. RESULTS: During the exercise protocol, peak torque decreased 30% and 24% in the eccentric and concentric groups, respectively, whereas no change occurred in EMG root-mean-square (RMS). There were no group differences in peak torque generation at any of the postexercise protocol time points. Compared with the preexercise protocol values, multifidus EMG was elevated 27% immediately post and 15 min post in the eccentric group. Similarly, compared with the concentric group, multifidus EMG in the eccentric group was increased 34%, 40%, and 25% immediately post, 15 min post, and 1 d after the exercise protocol, respectively. CONCLUSION: Eccentric contractions using the trunk extensor muscles result in higher levels of multifidus EMG activity to produce a given level of torque. This reduction in neuromuscular efficiency persisted for one day with recovery to baseline levels by the third day. Contrary to studies using other muscle groups, no sustained alteration in muscle function was observed. PMID- 11404664 TI - Electromechanical delay in isometric actions initiated from nonresting levels. AB - PURPOSE: The purpose of this study was to determine whether electromechanical delay (EMD) was associated exclusively with the onset of tension from a resting state and whether EMD remained constant across different rates of force development. METHODS: Twenty-four subjects (23.9 +/- 5.4 yr, 171.7 +/- 7.3 cm, 72.9 +/- 12.8 kg) performed isometric elbow flexion trials in the transverse plane by using the dominant arm during which isometric force data and surface EMG activity were collected. Subjects completed three trials to establish a maximal force (MF) reference. Subjects then completed trials in which pulse forces of varying magnitudes were elicited at a frequency of 1 Hz from different baseline intensities. All forces were expressed relative to MF. Three trials of the following conditions (baseline-pulse) were performed in random order: 0-25%, 25 50%, 50-75%, 0-50%, and 0-75%. EMG and force data were collected for 10 pulse cycles during these trials. EMD was defined as the temporal shift that maximized a normalized cross-correlation function. RESULTS: EMD for a 25% pulse force developed from rest (83.5 +/- 12.9 ms) was significantly longer than that developed from 25% (66.3 +/- 11.5 ms) or 50% (60.6 +/- 16.6 ms) baselines. EMD values were not different when force was developed from 25% and 50% baselines. EMD associated with a 25% pulse force from rest was significantly longer than 50% (70.3 +/- 10.0 ms) and 75% (68.9 +/- 8.7 ms) pulse forces from rest. EMD for 50% and 75% pulse forces from rest were not statistically different. CONCLUSION: It was concluded that EMD is present during exertions initiated from both resting and nonresting states but is reduced when exertions are initiated from non resting states and with higher rates of force development. PMID- 11404665 TI - A three-dimensional biomechanical analysis of the squat during varying stance widths. AB - PURPOSE: The purpose of this study was to quantify biomechanical parameters employing two-dimensional (2-D) and three-dimensional (3-D) analyses while performing the squat with varying stance widths. METHODS: Two 60-Hz cameras recorded 39 lifters during a national powerlifting championship. Stance width was normalized by shoulder width (SW), and three stance groups were defined: 1) narrow stance squat (NS), 107 +/- 10% SW; 2) medium stance squat (MS), 142 +/- 12% SW; and 3) wide stance squat (WS), 169 +/- 12% SW. RESULTS: Most biomechanical differences among the three stance groups and between 2-D and 3-D analyses occurred between the NS and WS. Compared with the NS at 45 degrees and 90 degrees knee flexion angle (KF), the hips flexed 6-11 degrees more and the thighs were 7-12 degrees more horizontal during the MS and WS. Compared with the NS at 90 degrees and maximum KF, the shanks were 5-9 degrees more vertical and the feet were turned out 6 degrees more during the WS. No significant differences occurred in trunk positions. Hip and thigh angles were 3-13 degrees less in 2-D compared with 3-D analyses. Ankle plantar flexor (10-51 N.m), knee extensor (359 573 N.m), and hip extensor (275-577 N.m) net muscle moments were generated for the NS, whereas ankle dorsiflexor (34-284 N.m), knee extensor (447-756 N.m), and hip extensor (382-628 N.m) net muscle moments were generated for the MS and WS. Significant differences in ankle and knee moment arms between 2-D and 3-D analyses were 7-9 cm during the NS, 12-14 cm during the MS, and 16-18 cm during the WS. CONCLUSIONS: Ankle plantar flexor net muscle moments were generated during the NS, ankle dorsiflexor net muscle moments were produced during the MS and WS, and knee and hip moments were greater during the WS compared with the NS. A 3-D biomechanical analysis of the squat is more accurate than a 2-D biomechanical analysis, especially during the WS. PMID- 11404666 TI - A physiological profile of tennis match play. AB - PURPOSE: The aim of this investigation was to examine physiological demands of single match play in tennis. METHODS: 20 players performed 10 matches of 50 min. Respiratory gas exchange measures (RGEM) and heart rates (HR) were measured using two portable systems. Lactate concentration was determined after each game. The average oxygen uptake (VO2) of 270 games was 29.1 +/- 5.6 mL.kg-1.min-1 (51.1 +/- 10.9% of VO2max). Average VO2 for a game ranged from 10.4 to 47.8 mL.kg-1.min-1 (20.4 and 86.8% of VO2max). Average lactate concentration (LA) was 2.07 +/- 0.9 mmol.L-1 (ranging from 0.7 to 5.2 mmol.L-1). Furthermore, we monitored the duration of rallies (DR), the effective playing time (EPT), and the stroke frequency (SF). The average values of 270 games were DR: 6.4 +/- 4.1 s, EPT: 29.3 +/- 12.1%, SF: 42.6 +/- 9.6 shots.min-1. RESULTS: Multiple regression revealed that the DR was the most promising variable for the determination of VO2 in match play (r = 0.54). The body surface area (BSA) and EPT were also entered into the calculation model. In games of two defensive players, VO2 was significantly higher than in games with at least one offensive player. CONCLUSION: Our results suggest that energy demands of tennis matches are significantly influenced by DR. The highest average VO2 of a game of 47.8 mL.kg-1.min-1 may be regarded as a guide to assess endurance capacity required to sustain high-intensity periods of tennis matches compared with average VO2 of 29.1 mL.kg-1.min-1 for the 270 games. Our results suggest that proper conditioning is advisable especially for players who prefer to play from the baseline. PMID- 11404667 TI - Enhancing cycling performance using an eccentric chainring. AB - PURPOSE AND METHODS: This study was designed to compare the physiological responses and performance of well trained cyclists riding with two different chainring designs, round or eccentric, during a brief and intense cycling exercise: an "all-out" 1-km laboratory test. The eccentrically designed chainring was made of two crank arms sliding into each other, with the inside arm fixed on the center of the arm of a circular chainring and the outside arm sliding along the inside and revolving around an elliptical cam. This design increases crank arm length at the downstroke and decreases it during the upstroke, thus increasing and decreasing the torque. In terms of the chainring's revolution, the crank arm length at 0 degrees and 180 degrees is similar to the arm length of circular chainrings (175 mm). However, during the downstroke (0-180 degrees ), it increases to its maximum length of 200 mm at 90 degrees and then returns to its original length of 175 mm at 180 degrees. During the upstroke, it decreases to a minimum length of 150 mm at 270 degrees and then increases to 175 mm at 360 degrees. Eleven cyclists performed an all-out 1-km laboratory test using each chainring. The study was conducted over two consecutive weeks with the order of chainring use randomized. During all trials, ventilatory data were collected every minute using an automated breath-by-breath system. Heart rate was measured using a telemetry system. RESULTS: None of the cardiorespiratory variables showed significant differences between chainring trials. Performance, however, was significantly improved using the eccentric design (64.25 +/- 1.05 vs 69.08 +/- 1.38 s, P < 0.004, with the eccentric and the round design, respectively). CONCLUSION: We concluded that the eccentric chainring significantly improved the cycling performance during an all-out 1-km test. Further testing with indoor cycling specialists performing on a velodrome would be helpful to define the maximal possibilities of such a chainring. PMID- 11404668 TI - Effect of resistance training on women's strength/power and occupational performances. AB - PURPOSE: The effects of resistance training programs on strength, power, and military occupational task performances in women were examined. METHODS: Untrained women aged (mean +/- SD) 23 +/- 4 yr were matched and randomly placed in total- (TP, N = 17 and TH, N = 18) or upper-body resistance training (UP, N = 18 and UH, N = 15), field (FLD, N = 14), or aerobic training groups (AER, N = 11). Two periodized resistance training programs (with supplemental aerobic training) emphasized explosive exercise movements using 3- to 8-RM training loads (TP, UP), whereas the other two emphasized slower exercise movements using 8- to 12-RM loads (TH, UH). The FLD group performed plyometric and partner exercises. Subjects were tested for body composition, strength, power, endurance, maximal and repetitive box lift, 2-mile loaded run, and U.S. Army Physical Fitness Tests before (T0) and after 3 (T3) and 6 months of training (T6). For comparison, untrained men (N = 100) (MEN) were tested once. RESULTS: Specific training programs resulted in significant increases in body mass (TP), 1-RM squat (TP, TH, FLD), bench press (all except AER), high pull (TP), squat jump (TP, TH, FLD), bench throw (all except AER), squat endurance (all except AER), 1-RM box lift (all except aerobic), repetitive box lift (all), push-ups (all except AER), sit ups (all except AER), and 2-mile run (all). CONCLUSIONS: Strength training improved physical performances of women over 6 months and adaptations in strength, power, and endurance were specific to the subtle differences (e.g., exercise choice and speeds of exercise movement) in the resistance training programs (strength/power vs strength/hypertrophy). Upper- and total-body resistance training resulted in similar improvements in occupational task performances, especially in tasks that involved upper-body musculature. Finally, gender differences in physical performance measures were reduced after resistance training in women, which underscores the importance of such training for physically demanding occupations. PMID- 11404669 TI - The effect of three different warm-up intensities on kayak ergometer performance. AB - PURPOSE: The purpose of this study was to investigate the influence of warm-up (WU) intensity on supramaximal kayak ergometer performance. METHODS: In the initial testing session, eight institute of sport kayak squad members performed a graded exercise test for determination of VO2max and lactate (La) parameters. In a random, counterbalanced order, subjects subsequently performed WU for 15-min at either their aerobic threshold (W1), their anaerobic threshold (W3), or mid-way between their aerobic threshold and anaerobic threshold (W2). A 5-min passive rest period and then a 2-min, all-out kayak ergometer test followed the WU. RESULTS: For the three different WU conditions, no significant differences were observed for average power, peak VO2, total VO2, total VCO2, or accumulated oxygen deficit (AOD) during the 2-min test. However, when compared with W3, differences in average power approached significance after both W1 (P = 0.09) and W2 (P = 0.10). Furthermore, when compared with W3, average power during the first half of the 2-min test was significantly greater after W2 (P < 0.05) and approached significance after W1 (P = 0.06). After each WU period, there was a significant difference in blood pH (W1>W2>W3; P < 0.05) and blood [La] (W110 cc; p=0.019), sex (84.2% for boys and 40% for girls), of angioarchitecture and Dmin (p=0.01). After multivariate analysis, only Dmin was nearly correlated with the obliteration rate (b=0.462 7; SE=0.244 0; p=0.057). Actuarial risk of hemorrhage was 2.12% per year per patient. Patients with seizures were seizure-free without any drugs after radiosurgery in 80% of cases. Parenchymal changes were mainly hypersignals spT2 in MRI. CONCLUSION: Radiosurgery seems to be an efficient treatment with a low morbidity for cerebral arteriovenous malformations in children. PMID- 11404710 TI - [Retrospective analysis of failures in radiosurgery treatment of cerebral arteriovenous malformations]. AB - The aim of this paper is to analyze retrospectively the reasons for the failure in cerebral arteriovenous malformations radiosurgery. Several factors are evoked and discussed mainly: inaccurate target, intentional partial irradiation, repermeabilization of a previously embolized cerebral arteriovenous malformation. The results suggest the necessity of a complete irradiation of the nidus. The strategy of partial volume irradiation should be avoided, even if it necessitates lowering of the doses in large cerebral arteriovenous malformations. Accuracy in the target determination is required and a complete stereoangiography is necessary. PMID- 11404711 TI - [Surgical removal of radio-induced lesions after radiosurgery of cerebral arteriovenous malformations]. AB - Radioinduced lesions after radiosurgery of cerebral arteriovenous malformations may be associated with an increased signal on T2-weighted and gadolinium enhancement on T1-weighted MR images. They do not have necessarily a poor prognosis. These lesions are mostly asymptomatic. But in a few cases they can be associated with severe clinical symptoms which can become corticodependant or corticoresistant. We present the 5 cases of such cerebral arteriovenous malformations treated by radiosurgery, out of our series of 705 patients. The removal was easier than that of untreated cerebral arteriovenous malformations, and led to a complete recovery of symptoms and progressive decrease of imaging abnormalities. Such surgery should be proposed in case of symptomatic radioinduced lesions which fail to respond to steroids. PMID- 11404712 TI - [Re-irradiation of cerebral arteriovenous malformations. Experience of the Sainte Anne-Tenon Group]. AB - BACKGROUND AND PURPOSE: Between 20 to 50% of cerebral arteriovenous malformations treated with radiosurgery (RS) fail to obliterate 2 to 5 years after irradiation. Patients are not protected against the risks leading to treatment. Two therapeutic options can be used to eradicate the persisting nidus: micro-surgery and a second irradiation. Our group has reirradiated 39 such patients. MATERIAL: From 1989 to 2000, 39 patients have been reirradiated (14 females and 25 males; median age 31 years). There were more left lesions: 59% than right (35%) and 5% on midline. The most frequent locations were: temporal 12 cases; parietal 8 cases; frontal 7 cases; thalamus 7 cases. The predominant first symptoms were hemorrhage (68.5%) and seizure (15.8%). Prior RS, 21/39 patients had embolization (53.8%) and 3 surgery. Method. Treatment has been performed with the same system for the first and the second radiosurgery for 37 patients. Planification and dosimetry improved during that period. The level of dose was similar for the 2 RS. MRI has been used as a non invasive follow-up tool. RESULTS: Only 28 patients were evaluable because 7/39 patients had the second radiosurgery in 1999 or in 2000 and data were lacking at the time of writing for 4 patients. Obliteration rate was 17/28 (60.7%). Nine patients bled between the two radiosurgery procedures. COMPLICATIONS: 4 new regressive deficits occurred after the second radiosurgery. The rate of parenchymal changes were higher, after the second radiosurgery. Except one patient who died of a non-related affection 2 years after obliteration of his cerebral arteriovenous malformation, thus 38/39 patients were alive. CONCLUSION: This series was small compared to the potential number of candidates suffering from failure of the first radiosurgery, but the results are promising. PMID- 11404713 TI - [Second treatment of cAVMs after radiosurgery]. AB - Risks of bleeding from partially or unobliterated cerebral arteriovenous malformations remain unchanged. A complementary treatment should be indicated after radiosurgery. In this brief review the following data are discussed: the reasons of second treatment, imaging follow-up of cerebral arteriovenous malformations after radiosurgery, types of cerebral arteriovenous malformations to be treated, timing and therapeutic modalities. PMID- 11404714 TI - [Hemorrhage after radiosurgery of cerebral arteriovenous malformations]. AB - BACKGROUND AND PURPOSE: Obliteration is progressive after radiosurgery of cerebral arteriovenous malformations. Thus the hemorrhage risk still remains until obliteration. Purposes of this study are to appreciate severity of post radiosurgery hemorrhages, actuarial risk of hemorrhage and parameters associated with it. PATIENTS: and method. Over 705 patients treated, 46 (6.5%) had one or several hemorrhages. Clinical, anatomic, dosimetric parameters and obliteration rates before hemorrhage were studied. Then, actuarial risks per patient and per hemorrhage were calculated. Correlations between parameters and risk were searched by uni and multivariate analysis by drawing hemorrhage-free survival curves (limit-product Kaplan-Meier) and Cox model. RESULTS: Except one pure ventricular hemorrhage causing death of one patient, only parenchymal hemorrhages were associated with morbidity (80% of cases with 45% of permanent deficits). Overall mortality rate by hemorrhage was 6.5%. Overall morbidity rate was 34.8% and 13.6% for permanent deficit. Mean obliteration rate before hemorrhage was 25%. Actuarial hemorrhage rate were 2.98% per patient and 3.24% per hemorrhage. Actuarial rate per patient increased from 1.46% first year to 5.95% 4(th) year after radiosurgery. Parameters correlated with hemorrhage risk were in univariate analysis size (p=0.01), Spetzler and Martin's grade (p<0.001), dose to reference isodose (p=0.03), Dmin (p=0.08), intra or paranidal aneurysms (p<0.001), and recoverage (p<0.001). After multivariate analysis, only intra or paranidal aneurysms, recovering and Dmin were significantly associated with hemorrhage-free survival after RS. CONCLUSIONS: Post-radiosurgery hemorrhages are often sum of hemorrhage risk factors of the cerebral arteriovenous malformation and factors predicting low rate of obliteration. They can be in some cases foreseen but rarely avoided. PMID- 11404715 TI - [Evolution of epileptic seizures associated with cerebral arteriovenous malformations after radiosurgery]. AB - The goal of this study is to evaluate the response of epileptic seizures associated with cerebral arteriovenous malformations following radiosurgery and determine the factors associated with a positive outcome. The series included 210 patients (123 men, 87 women) with a mean age of 33. One hundred seventy three patients out of 210 were controlled after radiosurgery. Seizures completely disappeared in 58% of cases and decreased in 18%. Anti-epileptic treatment was stopped in 44%. Sex and age upon seizure appearance were not found as determining factors in patients whose seizures disappeared after radiosurgery. Generalized seizures disappeared following radiosurgery in 90.7%, partial secondary generalized seizures in 53.5%, and partial seizures in only 39.6%. Frontal or temporal malformations are associated with a better response after radiosurgery than sylvian or rolandic location. Seizure disappearance was maximal in case of isolated seizure prior to the radiosurgery (84.4%), was reduced to 61.2% when seizures were limited to 2 to 20, and disappeared in 21.4% in severe seizures. Epilepsy present less than 3 years prior to radiosurgery disappeared in 71.9%; was reduced to 45% when the delay was over 3 years. Seizures disappeared in 64.4% when the malformation was obliterated, they were reduced to 46.9% when malformations persisted. The persistence of severe seizures following radiosurgery was linked to persistence of the malformation. On the other hand early seizures occurred in 3.3% during the first 48 hours following radiosurgery; they disappeared in 72.2% after that period. De novo seizures also occurred in 1.7% and disappeared in all these cases. In conclusion, seizures associated with cerebral arteriovenous malformations respond well to radiosurgery; the malformation obliteration evokes its association with the seizure; the seizure disappearance in spite of persistence of the malformation evokes the positive effect of radiotherapy on epileptic seizures. PMID- 11404716 TI - [Course of headaches associated with cAVMs after radiosurgery]. AB - The purpose of this study is to evaluate the response of headaches associated with cerebral arteriovenous malformations following radiosurgery and determine the factors that are associated with a positive outcome. From our series of 700 patients whose cAVM was treated by radiosurgery, this study includes 109 patients (48 males, 61 females) with a mean age of 32 when radiosurgery was performed. Headaches resolved in 37 patients (53.7%), decreased in 14 patients (20.3%), persisted in 16 patients (23.2%), and even increased in frequency in 2 (2.9%). Headaches disappeared 2 years following radiosurgery in the majority of patients (31 out of 37: 83.3%). The disappearance rate of headaches was better when it was the only sign of the malformation (64%) or preceded an epileptic seizure (60%) with respect to the case when it preceded cerebral hemorrhage (34.8%). Headaches having the best response were those with 1 to 2 episodes per month (64.9%) and whose episode duration was less than 3 hours (67.6%). Unilateral headaches accounted for 86.5% of resolved headaches whereas bilateral headaches were only 8.1% and even less were the diffuse ones (5.4%). The best evolution was obtained for those associated with a parietal location of the cerebral arteriovenous malformation, followed by rolandic (respectively 55.5% and 50%). Occipital malformation was associated with headache disappearance in 38.1% and temporal with 34.3%, whereas frontal location was associated with headache disappearance in only 25% of the cases. Headache disappearance was often correlated with cerebral arteriovenous malformation outcome after radiosurgery (64.9% of disappearance in case of complete obliteration). However, in 24.8%, headaches disappeared prior to the malformation obliteration and were associated with a decrease of the flow, of the nidus volume or with the nidus disappearance with the persistence of a precocious venous drainage. Headaches disappeared in spite of the malformation persistence in 5.4% of the cases. We found de novo headaches in one patient one year following RS. Another patient presented an episode of headache 12 hours following RS. The course of headaches associated with cerebral arteriovenous malformations after radiosurgery is positive in the majority of cases and correlated to the malformation outcome. This correlation suggests a link between cerebral arteriovenous malformations and headaches and the equal benefit of radiosurgery on both of them. PMID- 11404717 TI - [Parenchymal changes after radiosurgery of cerebral arteriovenous malformations. Clinical and MRI data]. AB - BACKGROUND AND PURPOSE: Purposes of this study are to describe different parenchymal changes seen after radiosurgery of cerebral arteriovenous malformations and the clinical symptoms which can be associated, and risk factors correlated with them. PATIENTS: and method. From the whole population of 705 patients with a cerebral arteriovenous malformations treated by radiosurgery between 1984 and 1998, clinical from 615 patients and post radiosurgery MRI data from 367 patients were reviewed. Neurological deficit occurred in 5.37% of cases and was still persistant in 1.46% of cases. Delay of occurrence of deficits ranged from 6 to 83 months (mean: 27 months, median: 15 months). Parenchymal changes seen in MRI were classified in 4 grades: 1 without parenchymal changes, 2 hypersignal in sp T2, 3=2 with homogenous enhancement with gadolinium, 4 with hyposignal in spT1 and annular irregular enhancement. Several parameters (size, volume, angioarchitecture of the cerebral arteriovenous malformation, dosimetric parameters) were studied and correlations were searched by uni and multivariate analysis with occurrence and delay of occurrence of deficits or parenchymal changes. RESULTS: In multivariate analysis, only size was significantly correlated with occurrence of parenchymal changes (p=0.0016); only size of the malformation was significantly correlated with delay of occurrence of parenchymal changes (p=0.0082); only grade 4 was correlated with occurrence of neurological deficit (p<0.00001). However, when only "a priori" parameters (known before radiosurgery) are introducted in logistic model, size taille (p=0.02) and hypoplasy of a sinus (p=0.0049) are significantly correlated with occurrence of neurological deficit. Only parenchymal changes grade 4 was significantly correlated with delay of occurrence of a neurological deficit (p<0.00001). However, when only a priori parameters (known before radiosurgery) are introducted in logistic model, only arterial steal (p=0.054) was significantly correlated with delay of occurrence of a neurological deficit. CONCLUSION: Parenchymal changes are various in expression, signification and clinical symptoms associated with them. They must be known and recognized for better prevention and symptomatic treatment as well. PMID- 11404718 TI - [Postoperative morbidity and mortality after microsurgical exclusion of cerebral arteriovenous malformations. Current data and analysis of recent literature]. AB - BACKGROUND AND PURPOSE: Microsurgical exclusion of a cerebral arteriovenous malformation (AVM) can compare favorably with radiosurgery. We sought to assess its rate of morbidity-mortality as it is presently reported in the literature, and to discuss some of its current and worthwhile indications. METHODS: Through Medline and additional searches by hand, we retrieved studies reporting the clinical and angiographic results after microsurgical excision of an AVM published between january 1990 and december 2000. RESULTS: a) Postoperative mortality was 3.3% (68/2 452 patients from 25 studies). Permanent postoperative morbidity was 8.6%. Morbidity was never absent varying from 1.5% to 18,7%. The morbidity-mortality rate increased with an increasing Spetzler-Martin's grade, and was related to the location of the AVM. A 4.6% morbidity (from 1.5% to 9.7%) and a zero mortality were reported after microsurgical removal of small lesions of less than 3 cm in diameter. b) Postoperative angiography confirmed a total excision of the AVM in 97% of the cases (1 050/1 076 patients over 11 series), varying from 91% to 100%. c) Permanent morbidity related to pre-surgical embolization varied from 4% to 8.9%. Results from multiple or combined treatment including microsurgery could not be summed up. CONCLUSIONS: A complete and definitive microsurgical excision of an AVM can be achieved with a high success rate and a low morbidity-mortality rate, according to sound indications and to the neurosurgeon's personal experience. The choice for a best treatment of an AVM is no longer limited to microsurgery; it is a team decision where the neurosurgeon plays a determining role. PMID- 11404719 TI - [Embolization of cerebral arteriovenous malformations]. AB - Embolization is an important therapeutic tool for the treatment of cerebral arteriovenous malformations and is a part of the therapeutic strategy. In selected cases, it may lead to a total and permanent cure per se, but in most cases it will be an adjunctive therapy to radiosurgery. The authors review techniques of embolization, perioperative care, complications and applications of the treatment. PMID- 11404720 TI - [Treatment of cerebral arteriovenous malformations: decision making strategy]. AB - We have evaluated our therapeutic strategy through the results of modalities used alone or in association. Should the place of each modality be modified? What could be the influence of the first consulting physician? We feel at the present time that small cerebral arteriovenous malformations could be treated by radiosurgery, surgery and embolization according to their characteristics and location. For middle size and some large cerebral arteriovenous malformations, we consider that embolization and/or surgery are adjuvant modalities, specifically when the cerebral arteriovenous malformation is located in an eloquent area. PMID- 11404721 TI - [Role of Helicobacter pylori infection in insulin-dependent diabetes mellitus]. AB - BACKGROUND: Patients with insulin-dependent diabetes mellitus (IDDM) are often affected by chronic infections. H. pylori is one of the most common infection worldwide. Aim of the paper was to evaluate the prevalence of H. pylori infection, the efficacy of the most common H. pylori eradication therapies and the incidence of H. pylori reinfection after 12 months follow-up in IDDM and in a control group of dyspeptic patients matched for sex and age. METHODS: 116 IDDM patients and 50 dyspeptic controls were evaluated for H. pylori infection through 13C-urea breath test (UBT). 64 IDDM infected patients and 50 dyspeptic infected controls were randomly assigned to 3 different 7-day eradication regimens: 1) amoxicillin, clarithromycin, pantoprazole; 2) tinidazole, clarithromycin, ranitidine bismuth citrate; 3) tinidazole, clarithromycin, pantoprazole. Patients not eradicated by the first cycle were subsequently submitted to a 7-day quadruple therapy with tinidazole, tetracycline, bismuth and pantoprazole. Gastrointestinal symptoms and side effects were evaluated. Thirty-four IDDM patients and 40 dyspeptic patients previously treated for H. pylori infection and successfully eradicated (confirmed both by UBT and histology) were re-evaluated after 12 months. RESULTS: 37% of IDDM patients were infected. Mean age of IDDM infected patients was higher than negative (40+/-12 vs 32+/-10 yrs). None of the triple therapies used allowed an eradication higher than 62%. Conversely, the quadruple regimen eradicated 88% of patients. Minor side effects were observed in 10% of patients submitted to the triple therapies and in 25% of patients treated with the quadruple therapy (p<0.05). H. pylori re-infection was significantly higher in IDDM patients compared to controls: (38% vs 5% respectively, p<0.001). Mean age of IDDM re-infected patients was higher than negative (40+/-12 vs 32+/ 10). Among IDDM patients daily insulin requirement and glicate haemoglobin were significantly higher in re-infected compared to uninfected patients (44+/-9 vs 35+/-8 U/I and 7,2+/-1 vs 6,8+/-0,8). CONCLUSIONS: Prevalence of H. pylori infection in IDDM patients was similar to controls. IDDM patients show a low H. pylori eradication rate with a standard triple therapy regardless of the regimen utilized. The use of a quadruple regimen leads to the cure of a large percentage of the infected patients not eradicated by the first therapy, although it is accompanied by a greater incidence of minor side effects. A significantly higher incidence of H. pylori re-infection was observed in IDDM patients when compared to not IDDM controls. The data suggest that vaccine development seems to be the only effective long run treatment for this subset of patients. PMID- 11404722 TI - [The role of tetracycline in the retreatment after Helicobacter pylori eradication failure]. AB - BACKGROUND: To study the effect of second-line treatment with tetracycline (T) combination therapy in patients with Helicobacter pylori (H. pylori) infection after failure of triple one-week therapy with a proton pump inhibitor, omeprazole (O) with amoxycillin (A) and claritromycin (C) or metronidazole (M). METHODS: Three hundred twenty-five naive patients (146 males, 179 females, mean age 50, range 18-76), with H. pylori infection, were randomised to receive one-week triple therapy b.i.d. with O 20 mg; A 1 g; C 500 mg (OAC7 163 pts) or O 20 mg; A 1 g; M 500 mg (OAM7 162 pts). H. pylori status was determined before therapy by histology and 3 months after the end of treatment by C-13 urea breath test (UBT). When eradication did not occur, the patients were retreated with OTA7: O 20 mg b.i.d.; T 500 mg t.i.d. and A1 g b.i.d. for another week. H. pylori was assessed by C-13 UBT 3 months after the end of retreatment. Statistical analysis was done by c2 and Student t test. RESULTS: Three hundred and ten patients have completed the study with compliance >95%: 15 patients complained of side effects (diarrhea, nausea, urticaria, abdominal pain): 9 group OAC7, 6 group OAM7. After the first treatment h. pylori eradication were obtained in 124/154 (80.5%) (PP) OAC7 group and 123/156 (78.8%) (PP) OAM7 group (ns). 49/63 patients (77.7%) were negative after second-line treatment with OTA7 regimen. CONCLUSIONS: In personal experience the drug compliance was excellent. The rate of eradication after one week therapy-omeprazole combined with A+C or with A+M-was satisfactory. The retreatment with tetracycline combination regimen cured 77.7% of patients: that seems to be a promising options, in clinical practice, after an eradication failure. PMID- 11404723 TI - [Observations on the increased serum levels of YKL-40 in patients with rheumatoid arthritis and osteoarthritis]. AB - BACKGROUND: This study aimed to evaluate the function of glycoprotein YKL-40 in patients with rheumatoid arthritis, erosive osteoarthrosis or coxitis. METHODS: Serum levels of glycoprotein YKL-40 were assayed using an ELISA method in 40 patients with active-phase rheumatoid arthritis and 20 patients suffering from erosive osteoarthrosis or coxitis. RESULTS: The mean values of YKL-40 were enhanced in all patients with rheumatoid arthritis and arthrosis with higher mean values in erosive osteoarthritis and coxitis (440 ng/ml) compared to rheumatoid arthritis (190 ng/ml). CONCLUSIONS: YKL 40 is a local prognostic marker of joint destruction. PMID- 11404724 TI - [Determination of opiates in urine: interpretation of comparison between the EIA and FPIA and confirmation of data with GC/MS]. AB - BACKGROUND: The aim of this study was to evaluate some urine samples and to analyse them using two immunochemical screening methods. METHODS: A more reliable method has been used to obtain a critical analysis of the validity of the first analysis methods used. A total of 147 urine samples were examined using both the FPIA technique (Adx, Abbott) and the EIA technique (Random 120, Bracco); confirmation was obtained using GC/MS. RESULTS: When the threshold was altered from 200 to 300 ng/ml the EIA technique is more severely affected by changes in reference values compared to FPIA. In different values between two techniques, confirmation with GC/MS was possible only 5 cases. CONCLUSIONS: The study of data processing with immunochemical tests and after confirmation showed that the results with GC/MS are closer to Adx. The FPIA results are in agreement with the GC/MS technique both at 200 and 300 ng/ml with a percentage of 80.0%. The EIA technique has a different result, at 200 ng/ml it is in agreement for 40.0%, but at 300 ng/ml there is only 20.0%. PMID- 11404725 TI - [Gait in patients with Parkinson's disease after surgically treated hip fracture]. AB - BACKGROUND: The aims of this study were: 1) to evaluate the degree of walking of patients with Parkinson s syndrome after hip fracture surgically treated; 2) to reexamine the evidence for internal fixation versus endoprosthesis. METHODS: Forty-one patients with Parkinson s syndrome after hip fracture surgically treated have been studied. Twenty-four fractures (20 femoral neck, 4 subcapital) were treated by endoprosthesis, and 17 by internal fixation (pertrochanteric fractures). The average age of the patients was 76+/-7 years (7 male, 34 female); 97,5% patients could walk without any help before the fracture. RESULTS: At the end of the rehabilitation period, 21.9% patients were unable to walk; 2.4% walked with assistance of two persons, 24.4% with assistance of one person, 19.4% with two canes, 17.1% with one cane, 14.8% without assistance and without cane. At discharge, 87.5% of patients went directly back home, while the remaining 12.5% were admitted to Geriatric hospitals. The degree of walking was higher in fractures treated by endoprosthesis (p<0.05). At the end of treatment, 62.5% fractures treated by endoprosthesis and 35.2% treated by internal fixation could walk without any help. CONCLUSIONS: IN CONCLUSION: 1) Parkinson s disease influences the recovery of walking after hip fracture; 2) At the end of rehabilitation, the parkinsonian patients with hip fractures treated by endoprosthesis showed better results than those treated by internal fixation. PMID- 11404726 TI - Nitric oxide. A general review about the different roles of this innocent radical. AB - Nitric oxide, a short half-life radical, is highly reactive, and it is involved in many biological processes, such as vascular homeostasis, neurotransmission, and inflammation, defined as a sequence of events which can be simplified as follows: vasodilatation, alteration of vascular permeability, emigration of leucocytes from vessels, migration of leucocytes into the sites of tissutal damages or inflammation, activation of leucocyte mechanisms. This review has a double purpose: 1) to provide a comprehensive table of cell types that produce NO, together with the effects of agents used to study iNOS regulation; 2) to investigate the role of NO in different human systems. The different relations between NO and cytokines, the heart, infectious diseases, inflammatory diseases, brain cells and, lastly, gastrointestinal diseases are examined. PMID- 11404727 TI - [Idiopathic pulmonary fibrosis. A case report]. AB - A case of idiopathic pulmonary fibrosis is presented. The difficulty of a differential diagnosis with other interstitial lung diseases and the necessity of a right and complete diagnostic pathway are underlined. PMID- 11404728 TI - [Hyperkeratotic Kaposi sarcoma with leg lymphoedema after prolonged corticosteroid therapy for SLE. Case report and review of the literature]. AB - Kaposi sarcoma (KS) is a malignant vascular neoplasia with a viral etiology, characterized by development of multiple hyperpigmentate lesions, primarily at cutaneous level with associated edema and ulcerations, but frequently involving also the mucous membranes and/or visceral organs. In this study, we describe (in the light of the relevant literature) the clinical case of an elderly (78 yrs old) woman, who developed red-blues multiple hyperkeratotic nodules in the right leg and foot with marked lymphoedema, blushing and pain, after a long period of a low-dose corticosteroid therapy for LES (at least 10 years of continuous treatment). The diagnosis of KS was made on the basis of histologic findings. The patient HLA-typing showed the haplotypes HLA-A2-10, -B21-35, -Bw4-6, -Cw4 and HLA DR11-13 (some of which are known to predispose to LES, but not to KS). The KS, first described by Moritz Kaposi in 1872, has been a very rare pathology until the 80s, afterwards its frequency has steadly increased, favored by immunosuppressive therapy for autoimmune diseases or tranplants and by immunodepression of AIDS. Concerning the pathogenesis, it is crucial the role of HHV-8 of the herpesvirus family (found in the lesions and in the circulating cells of all KS patients), for which a prevailing sexual transmission is postulated. General physicians and specialists of internal medicine and angiology should know this disease, which can be undiagnosed because of the low incidence in the general population and the consequent poor knowledge of this vascular neoplastic disease, which is now reported with increasing frequency. PMID- 11404729 TI - [Cardiac involvement in Kawasaki disease. Our experience]. AB - BACKGROUND: Kawasaki disease is an acute vasculitis of unknown cause that affects children under 5 years of age and in 20-25% of cases can cause coronary artery anomalies. Early therapy with intravenous gamma globulin and aspirin greatly decreases the risk of developing coronary anomalies, but sometimes coronary disease develops even in timely diagnosed and treated cases. METHODS: Personal experience in 41 children with Kawasaki disease is reported. Age was within three months and 5 years. All patients had electrocardiography and echocardiography at diagnosis, after 1 week, at the third, sixth, and eighth week, after six months and after 1 year from diagnosis. Echocardiographic cardiac anomalies were found only in 5 cases (12.19%). Average age in these cases was lower. RESULTS: In these cases coronary involvement developed in spite of early diagnosis and therapy (intravenous gamma globulin 400 mg/kg die and aspirin) electrocardiographic findings were not relevant or specific to coronary arteries anomaly. In three cases coronary anomalies regressed during the first year from diagnosis, the other two are still in the first year of follow-up. Nine children had a long term follow-up (6 to 9 years) and they were all normal except one with renal arteritis and another with slight mitral incompetence. CONCLUSIONS: The authors underline the role of clinical and instrumental follow-up that should be extended to all patients affected by Kawasaki disease with coronary anomalies even if early regressed. PMID- 11404730 TI - Multicystic dysplastic kidney and contralateral vesicoureteral reflux. Renal growth. AB - BACKGROUND: To evaluate if vesicoureteral reflux (VUR) contralateral to the multicystic dysplastic kidney can interfere with the compensatory renal hypertrophy. METHODS: Twenty-seven patients (17 males, 10 females) with multicystic dysplastic kidney (MDK) (14 on the right, 13 on the left) have been treated at the Nephrology Unit of the Pediatric Department of the University of Verona from birth up to the second year of life. All these patients were diagnosed as having MDK by prenatal ultrasonography. Seven children (4 males and 3 females) had VUR (5 monolateral, 2 bilateral), diagnosed at the end of the first month of life. After diagnosis children underwent antibiotic prophylaxis with beta-lactam compounds at low doses. Four patients underwent a surgical correction of VUR associated with nephrectomy within the second year of life. The remaining 3 patients were treated with antibiotic prophylaxis; a progressive resolution or downgrading of reflux grade took place respectively in 1 and in 2 of them. Only 6 children with MDK underwent nephrectomy. Renal growth was studied by serial echographic measurements of the longitudinal renal lenght (performed at birth, at 6 months, and at 2 years of life). RESULTS: Renal length was 5.68+/ 1.24 cm, 6.72+/-0.88 cm, 8.56+/-1.27 cm in children without VUR, respectively at birth, 6 months and 2 years of life. Renal length was 4.65+/-0.63 cm, 6.70+/-0.64 cm, 7.07+/-1.14 cm in children with VUR, respectively at birth, 6 months and 2 years of life. A statistically significant difference was observed between the two groups at birth (p<0.05) and at 2 years of life (p<0.01). CONCLUSIONS: The conclusion is that VUR contralateral to the MDK is associated with small kidneys and reduced renal growth both at birth and at 2 years of life. PMID- 11404731 TI - Respiratory tract infections caused by respiratory syncytial virus in children. Diagnosis and treatment. AB - Respiratory syncytial virus (RSV) is the most common viral cause of lower respiratory tract infection in infancy and young children. No effective treatment for RSV lower respiratory tract infection (RSV-LRTI) exists. Ribavirine initially was reported to be an effective anti-viral drug for RSV-LRTI. However, subsequently performed trials could not reproduce these positive results and based on the current available evidence there is no place of ribavirin in the routine treatment of RSV-LRTI. The use of nebulised bronchodilator therapy in RSV LRTI has been subject of many trials, with conflicting RESULTS. Although the individual patient may have some short term benefit of nebulised bronchodilators there does not seem to be a sufficient scientific base for the standard use of bronchodilator therapy in infants and children with RSV-LRTI. There is increasing evidence that RSV-LRTI is an immune-mediated disease and therefore corticosteroids may be an effective treatment. The results from efficacy trials have demonstrated that corticosteroids are not effective for patients with mild RSV infection. In contrast, there is some evidence suggesting that it may be beneficial in patients with more severe RSV-LRTI. Immunoprofylaxis with hyperimmune immunoglobulins and monoclonal antibody against the viral F-protein have been shown to be effective in the prevention of severe RSV-LRTI. From the results of the therapeutic efficacy trials, however, it is evident that immunoglobulins have no place in the treatment of RSV-LRTI. In conclusion, although RSV infections each year have a considerable socio-economic impact attempts to find an effective therapy have been quite unsuccessful so far. PMID- 11404732 TI - [Assessment and measurement of pain in infants and children]. AB - The assessment of global pain in infants and children involves the analysis of subjective, behavioural and physiological components, beyond the psychological, familiar and environmental factors that influence and modulate the manifestation of painful symptom. Therefore, researchers and clinicians need valid and reliable instruments to obtain a correct measure of pain. Although subjective indices carry out a fundamental role in the assessment of pain (instruments adjusted to the age of the child), behavioural and physiological indices should be used when intelligence deficit or serious physical handicaps exclude such possibility. Behavioural aspects, associated and correlated to physiological changes, allow to obtain a reliable enough measure of pain. In this paper, the main instruments for measuring pain in infants and children will be described, and their advantages and disadvantages underlined. PMID- 11404733 TI - [Experience of comic relief in pediatric hospital]. PMID- 11404734 TI - [Social assistance]. PMID- 11404735 TI - [Puericulture and pedagogy]. PMID- 11404736 TI - Congenital nasolacrimal duct obstruction. PMID- 11404737 TI - The development of the art and science of strabismology outside North America: Part II. PMID- 11404738 TI - Comparison of balloon dacryocystoplasty to probing as the primary treatment of congenital nasolacrimal duct obstruction. AB - PURPOSE: In children older than 18 months, primary probing procedures for congenital nasolacrimal duct obstruction (CNLDO) are thought to have lower rates of success. This study compares the results of primary probing to balloon dacryocystoplasty (DCP) in children stratified by age category. METHODS: In a retrospective chart review, 29 eyes with CNLDO that underwent balloon DCP in children older than 18 months were identified and age-matched to 29 eyes that underwent probing. The eyes were divided into 3 age categories: category 1 (18-24 months), category 2 (24-36 months), and category 3 (>36 months). RESULTS: Of the 29 eyes treated with balloon DCP (mean age, 37.1 months), 26 were successfully treated. Twenty-five of the 29 matched probed eyes (mean age, 31.1 months) were successfully treated, resulting in an overall success rate of 90% for balloon DCP and 86% for primary probing. Within each age category, the success rate varied but did not show an advantage to balloon DCP. The presence of crusting and expressible discharge from the puncta during preoperative evaluation predicted a successful probing (OR, 16; 95% CI, 1.3-192). CONCLUSION: Overall, balloon DCP did not appear to present an advantage as compared with primary probing as the initial treatment in these children. Primary probing has an impressive overall success rate that did not diminish in the children older than 36 months. PMID- 11404739 TI - Endoscopic dacryocystorhinostomy in children. AB - BACKGROUND: Persistent nasolacrimal duct obstruction (NLDO) often requires treatment by probing, intubation, or balloon dacryoplasty. Refractory cases have been managed by external dacryocystorhinostomy (DCR), which leaves a scar; however, this procedure is generally avoided in young children. Endoscopic DCR has been successfully performed in adults and described in children. We report the success of this procedure in a series of pediatric patients. METHODS: A retrospective review of all endoscopic lacrimal procedures performed in a 3-year period was undertaken. Seventeen children (22 ducts) with persistent NLDO after at least one failed probing, with or without silicone tube placement, underwent endoscopic DCR. Follow-up ranged from 6 to 36 months, and success was defined as resolution of tearing and discharge by follow-up clinical evaluation and by parental history. RESULTS: All but 2 patients (88%) with NLDO showed complete resolution of tearing and discharge. These 2 patients had recurrent symptoms after the Crawford tubes were removed and required revision endoscopic DCR. No complications from this procedure were noted. CONCLUSIONS: Endoscopic DCR is a safe and effective means of treating persistent NLDO in infants and young children when simple probing, intubation, or balloon procedures have failed. The team ophthalmology-otolaryngology endoscopic approach provides a highly successful alternative for patients with a persistent distal obstruction that might otherwise require an external procedure. PMID- 11404740 TI - The role of nasal endoscopy in repeat pediatric nasolacrimal duct probings. AB - PURPOSE: To determine whether pediatric nasal endoscopy improves treatment success in recurrent nasolacrimal duct obstruction. METHODS: A retrospective analysis was carried out on 2 nonrandomized cohorts of consecutively treated patients who had one failed nasolacrimal duct probing at the Hospital for Sick Children, University of Toronto. The first cohort of 24 consecutive patients (32 eyes), group A, was treated with Crawford silicone tube intubation by one ophthalmologist (R.C.P.). The second cohort of 23 consecutive patients (33 eyes), group B, was treated with repeat probing by a second ophthalmologist (A.V.L.) in conjunction with nasal endoscopy by one otolaryngologist (V.F.). Abnormalities found on endoscopy were treated accordingly, and no tubes were inserted. Follow up, through phone interviews or office visits, was conducted to assess the patients' symptoms. RESULTS: Group A patients were older at both the first probing (P = .048) and the second procedure (P = .012). No significant difference in the failure rates was found, with treatment failing in 3 eyes (2 patients) in group A and in 5 eyes (4 patients) in group B (P = .479). Interestingly, 17 of the 32 tubes in group A were extruded in 1 month or less. Also, 28 of 33 eyes in group B had abnormalities on endoscopy-some, multiple. Twenty-two eyes underwent inferior turbinate infracture: 5 had redundant mucosa, which was removed, and 6 had abnormal openings of the inferior meatus. CONCLUSIONS: We were unable to show any benefit of nasal endoscopy over intubation of the nasolacrimal system with silicone tubes in the treatment of failed probings despite the identification and treatment of abnormalities. The study was limited by its low power to detect differences because of the small number of patients and the high success rate of the traditional treatments for congenital nasolacrimal duct obstruction. PMID- 11404741 TI - The role of anisometropia in the development of accommodative esotropia. AB - PURPOSE: To determine whether anisometropia increases the risk for the development of accommodative esotropia with hypermetropia. METHODS: Records of all new patients with a refractive error of +2.00 D or more (mean spherical equivalent of both eyes) over a 42-month period were reviewed. Three hundred forty-five patients were thus analyzed to determine the effect of anisometropia (>or=1 D) on the relative risk of developing accommodative esotropia and of developing unsatisfactory control with spectacles once esotropia was present. RESULTS: Anisometropia (>or=1 D) increased the relative risk of developing accommodative esotropia to 1.68 (P < .05). Anisometropia (>or=1 D) increased the relative risk for esotropia to 7.8 (P < .05) in patients with a mean spherical equivalent less than +3.00 D and increased it to 1.49 (P < .05) in patients with a mean spherical equivalent of +3.00 D or more (P = .016). In patients with esotropia and anisometropia (>or=1 D), the relative risk for a deviation that was unsatisfactorily controlled with spectacles was 1.72 (P < .05) compared with patients with esotropia but without anisometropia. Unsatisfactorily controlled esotropia was present in 33% of patients with anisometropia versus 0% of patients without anisometropia, with a mean hypermetropic spherical equivalent of less than +3.00 D (P = .003); however, anisometropia did not significantly increase the relative risk of unsatisfactory control of esotropia with spectacles in patients with a hypermetropic spherical equivalent of +3.00 D or more. Although amblyopia and anisometropia were closely associated, anisometropia increased the relative risk for esotropia to 2.14 (P < .05), even in the absence of amblyopia. CONCLUSIONS: Anisometropia (>or=1 D) is a significant risk factor for the development of accommodative esotropia, especially in patients with lower overall hypermetropia (>+3.00 D). Anisometropia also increases the risk that an accommodative esotropia will not be satisfactorily aligned with spectacles. PMID- 11404742 TI - Unilateral recession and resection in Duane syndrome. AB - PURPOSE: To assess the efficacy of lateral rectus resection with medial rectus recession in the affected eye of patients with Duane retraction syndrome (DRS) with esotropia and limited abduction, compared with bilateral medial rectus recessions. METHODS: The charts of 9 patients with DRS who underwent a recession resection procedure and 10 patients with DRS who underwent bilateral medial rectus recessions were reviewed. Ocular ductions (graded from 0 = full duction to -4 = total deficit), severity of retraction, alignment, head position, and binocular single vision field (for study group only) were recorded before and after surgery. RESULTS: Before surgery, the study and control groups did not differ in mean primary position esotropia (16.9 and 18.8 PD, respectively), face turn (16.5 degrees and 15.0 degrees, respectively), average limitation of abduction (-3.9 and -3.7, respectively), or adduction (-0.1 and -0.3, respectively). After surgery, both groups had similar mean face turns (3.9 degrees and 1.0 degrees ), esotropia (3.3 PD and 1.0 PD), and abduction limitation in the affected eye (-2.4 and -2.6). However, mean adduction was significantly worse in the control group than in the study group (-1.5 vs -0.6, P = .02). Globe retraction improved in all control subjects. It worsened in 5 study subjects and did not improve in the other 4. In the study group, 1 patient required reoperation for undercorrection and another was overcorrected. CONCLUSION: Seven of 9 patients with DRS, selected on the basis of esotropia, limited abduction, and mild retraction, benefited from a recession-resection procedure. Abduction improved to the same degree as seen after bilateral medial rectus recessions, with less tendency to limit adduction. PMID- 11404743 TI - An adjustable superior oblique tendon spacer with the use of nonabsorbable suture. AB - PURPOSE: Philip Knapp described a method, sometimes referred to as the "chicken suture,"of securing a loose nonabsorbable suture to the cut ends of the superior oblique tendon to facilitate future reversal. The purpose of this study is to describe a modification of Knapp's technique to achieve partial, reversible, and intraoperatively adjustable superior oblique weakening. METHODS: The superior oblique tendon was exposed, 2 polyester nonabsorbable sutures were placed 4 mm apart, and the tendon was cut. With the use of a slip knot, the cut ends of the tendon were separated by 2 to 8 mm. Tendon separation was adjusted intraoperatively according to the exaggerated traction test and, in some cases, fundus torsion. Medical records of all patients who underwent surgery with this technique were reviewed and the outcomes tabulated. RESULTS: Twelve patients (16 eyes) were treated for superior oblique overaction and 3 patients (3 eyes) for Brown syndrome. Follow-up was 2 to 46 months (mean, 17 months). Mean superior oblique overaction improved from +1.3 before surgery to +0.3 after surgery, mean A pattern improved from 20 PD to 2 PD, and fundus intorsion improved from +1.2 to +0.3. In Brown syndrome, the mean elevation in adduction improved from -3.8 to 1.0. One patient from each group developed an overcorrection. None of the patients developed recurrence. The patients with Brown syndrome continued to improve over a 1-year period. CONCLUSIONS: The superior oblique tendon suture spacer is effective, intraoperatively adjustable, and technically easier to perform than a silicone expander procedure. This technique should be considered as an alternative for patients requiring superior oblique weakening. PMID- 11404744 TI - Torsion as a contributing cause of the anti-elevation syndrome. AB - BACKGROUND: The anti-elevation syndrome is an adverse outcome of anterior transposition of the inferior oblique muscle. The presumed cause is an excessive anti-elevating force vector that occurs with attempted elevation in abduction. This causes apparent overaction of the contralateral inferior oblique muscle due to fixation duress. It has been suggested that excessive residual extorsion may help explain this phenomenon. METHODS: Fundus photographs to assess torsion were evaluated by masked observers in 18 patients who had undergone anterior transposition of the inferior oblique muscle. Eight of the patients were found to have the anti-elevation syndrome and 10 were not. RESULTS: Patients with the anti elevation syndrome had more extorsion (mean, 16.6 degrees +/- 3.4 degrees ) than the patients who did not have the anti-elevation syndrome (mean, 8.8 degrees +/- 2.3 degrees ). This difference was significant (P < .0001). In addition, 2 patients who initially did not show the anti-elevation syndrome were found to have an increase in their fundus extorsion after they subsequently developed the anti-elevation syndrome. Two patients who had the anti-elevation syndrome showed a marked decrease in fundus extorsion after the anti-elevation syndrome was surgically eliminated by converting the anterior transposition to a simple recession. CONCLUSION: The presence of substantial extorsion may contribute to the cause of the anti-elevation syndrome after inferior oblique muscle anterior transposition. Lateral placement of the posterior (lateral) corner of the inferior oblique muscle at the time of surgery may cause substantial extorsion after surgery. PMID- 11404745 TI - Timing of postoperative adjustment in adjustable suture strabismus surgery. AB - PURPOSE: The use of adjustable sutures in strabismus surgery has increased the rate of surgical success. Little data are available on the optimum timing for postoperative adjustment after strabismus surgery. We wanted to compare 2 common practices of adjustable suture technique after strabismus surgery. METHODS: Two comparable groups of 40 patients each, who had strabismus surgery with adjustable suture technique, were prospectively studied. Group A had early adjustment the same day of the surgery about 6 hours after the operation, and group B had late adjustment the next day about 24 hours after the operation. Subjective scoring tables were used to evaluate the pain felt by the patient before, during, and after the adjustment and any difficulties of the adjustment process. Requirements of postoperative pain medications and final alignment 6 weeks after surgery were also compared. RESULTS: Despite adequate statistical power, no significant differences were found between the groups regarding pain before, during, and after adjustment, difficulties performing the adjustment, and final alignment after 6 weeks (P > .05). Both adjustment schedules were equally associated with mild to moderate pain before, during, and after the adjustment. In the first 24 hours after surgery, no overall difference in the use of pain medications was found. Nausea and vomiting in the first 24 postoperative hours were more common in the early adjustment group (P = .02). CONCLUSION: The surgeon can feel free to choose the timing for postoperative adjustment. However, when performing an early adjustment, the surgeon should be especially prepared to control nausea and vomiting. PMID- 11404746 TI - Evaluation of motor vehicle driving performance in patients with chronic diplopia. AB - BACKGROUND: Uncorrected diplopia is a condition that may make it unsafe for a person to operate a motor vehicle. In some jurisdictions, physicians are required by law to report the person with diplopia to the appropriate authority. METHODS: In this masked study, 10 patients of varying ages with stable diplopia of greater than 6 months' duration and 10 age-matched control subjects were placed in a driving simulator and evaluated on their performance. Various cues and threats, including near-accident situations, were presented; stimulus recognition and reaction times were recorded. RESULTS: No significant difference was found between the groups for either cue or threat recognition responses or reaction times. Increasing age was the factor most associated with poor response performance on all test measures (P < .001). Slowed response time in patients with poor binocular single vision was the only other significant association. CONCLUSION: Although response times were slower in subjects with poor binocular single vision scores, stimulus recognition responses were not significantly different; in our opinion, stimulus recognition is more relevant to driving performance and therefore chronic diplopia does not appear to be a contraindication for driving a motor vehicle. PMID- 11404747 TI - Surgical outcome after prism adaptation for esotropia with a distance-near disparity. AB - INTRODUCTION: Prism adaptation for patients with esotropia and a distance-near disparity is controversial. The purpose of this study was to evaluate the surgical outcome for patients who underwent prism adaptation for esotropia with a distance-near disparity and determine whether both preoperative sensory and motor fusion are necessary to determine surgical success. METHODS: The medical records of 65 prism-adapted patients with a distance-near disparity of 9 PD or more were reviewed. Prism responders had a fusion response to near Worth 4-dot test and a deviation with prisms of 8 PD or less of esotropia at near, and 5 PD or less of exotropia at distance, or both. The same criteria were used postoperatively to assess a successful surgical outcome. Patients were operated for the near angle or greater than the near angle. RESULTS: Fifty-eight of 65 patients (89%) demonstrated fusion with prisms. Twenty of 65 patients (31%) had increased deviations that were greater than their original near angle (prism builders). Eleven percent (7 of 65) had no fusion. Postoperatively, 88% (51 of 58) of all fusers, 95% (18 of 19) of prism builders, and 71% (5 of 7) of nonfusers had a good surgical outcome of 8 PD or less of esotropia at near, 5 PD or less of exotropia at distance, or both. CONCLUSIONS: Prism response for distance-near disparity esotropia is a good indicator of postoperative outcome. Responders to prism adaptation had a better surgical outcome compared with nonresponders. In particular, prism adaptation aids in detecting those patients who will benefit from larger amounts of surgical correction. PMID- 11404748 TI - Acquired esotropia: subjective and objective outcomes. AB - PURPOSE: The measurement of outcomes of pediatric therapy will be increasingly important to third-party payers as they allocate health care resources. We undertook this study to assess the effectiveness of treatment of acquired esotropia, as measured subjectively by parents and objectively by examination. METHODS: A chart review was used to gather objective clinical data from all patients presenting between 1994 and 1995 with esotropia after their first birthday. Parents' impressions of the quality and impact of treatment were tabulated by using a telephone questionnaire. RESULTS: Of 48 patients studied (mean age, 3.8 years; 44 months' follow-up), the 26 with amblyopia were effectively treated in 81% of cases. Mean esotropic angles decreased, with glasses and/or surgery, from 33.1 to 4.9 PD at distance and from 40.0 to 11.4 PD at near. The average estimated cost of care was 547 dollars per patient per year. Parents considered treatment "extremely important" to their child's future happiness and success in 90% of cases. CONCLUSION: Our experience has shown that treatment of acquired esotropia in childhood, while relatively inexpensive, is highly effective, both objectively and subjectively. PMID- 11404749 TI - Retinopathy of prematurity and weight of the baby's chart. PMID- 11404752 TI - The influences of partial denture design on distribution of force to abutment teeth. 1956. PMID- 11404753 TI - The use of implants in the occlusal rehabilitation of a partially edentulous patient: a clinical report. AB - This article describes the occlusal rehabilitation of a partially edentulous patient who did not want a removable partial denture. Implants and extensive fixed restorations were used to restore posterior support and treat severely worn dentition, respectively. The treatment offered the patient a functional and esthetic result. PMID- 11404754 TI - Retrospective assessment of 546 all-ceramic anterior and posterior crowns in a general practice. AB - STATEMENT OF PROBLEM: There is need to determine whether following a consistent procedural protocol in a private dental practice will result in high success rates for all-ceramic anterior and posterior crowns fabricated from aluminous porcelain veneered to high-alumina content-supporting cores. Aluminous porcelain is known to have poor tensile strength and fracture resistance when subjected to shear forces. PURPOSE: This study examined the long-term survivability of alumina core, all-ceramic crowns in both anterior and posterior locations. MATERIAL AND METHODS: Following a consistent protocol over 6 years, 546 all-ceramic, In-Ceram crowns (32.4% anterior [n = 177] and 67.6% posterior [n = 369]) were consecutively luted with glass ionomer resin hybrid cement and periodically observed by the same dentist who prepared and cemented them. RESULTS: The overall success rate was 99.1% (n = 541) with a 0.9% (n = 5) failure rate. The success-to failure ratio for anterior all-ceramic crowns was 98.9% to 1.1% (n = 175:2); the posterior crown ratio was 99.2% to 0.8% (n = 366:3). CONCLUSION: Following a consistent protocol of careful tooth preparation and crown cementation resulted in high success rates for anterior and posterior all-ceramic crowns. Glass ionomer resin hybrid cement appeared to be a reliable luting agent. All-ceramic alumina core crowns are durable and provide optimum esthetic choices for anterior and posterior locations. PMID- 11404755 TI - In vitro color stability of double-layer veneers after accelerated aging. AB - STATEMENT OF PROBLEM: Porcelain laminates made from thin alumina shells veneered with feldspathic porcelain could be a promising alternative to conventional veneers. Long-term shade stability is critical for esthetics. PURPOSE: This study compared changes in CIE L*a*b* color coordinates of simulated veneers made from aluminum oxide core material veneered with feldspathic porcelain after 300 hours of accelerated photothermal aging (weathering). MATERIAL AND METHODS: Fifteen aluminum oxide disks (Procera) were divided into 3 groups. Each of the 5 disks was veneered with All-Ceram porcelain of the Vita shades A1 and B4, respectively. Five disks remained unfinished as controls. The disks were bonded to composite substrates simulating stained teeth. The color of the specimens was measured with a colorimeter. All specimens were subjected to 300 hours of accelerated aging under light exposure and thermocycling. Color measurements were repeated, and the data were statistically evaluated with multiple paired t tests. RESULTS: Color changes in the test groups involved an increase in lightness and a decrease in chroma. The calculated total color differences were not statistically significant compared with a level of 3 DeltaE units. CONCLUSION: This simulation of double layer veneers appears to demonstrate acceptable color stability for this type of restoration. Clinical research is necessary to substantiate these in vitro findings. PMID- 11404756 TI - The effects of dowel design and load direction on dowel-and-core restorations. AB - STATEMENT OF PROBLEM: Complications such as loosening of the dowel and core or fracture of the remaining tooth root can be influenced by many factors, including the amount of remaining dentin, direction of the occlusal load, and design of the dowel. Most stress analyses of dowel and cores were conducted without including all aspects of the restorations and supporting structures. PURPOSE: The purpose of this study was to investigate the influence of occlusal stresses on various dowel designs in a restored, endodontically treated maxillary incisor by using a 2-dimensional finite element analysis model. MATERIAL AND METHODS: A 2 dimensional finite element model was constructed in a labiolingual cross sectional view of a maxillary central incisor, a dowel, a core, and the supporting tissues to investigate stresses in various dowel designs. As a control, a metal-ceramic crown on an endodontically treated tooth without a dowel and core was modeled. A 10-kg force was applied as follows: (1) in a vertical load on the incisal edge, (2) in a horizontal load on the labial surface, and (3) in a 20-degree diagonal load on the lingual surface. RESULTS: The use of a dowel reduced the peak dentinal stress to 75% of the magnitude of the control. When a vertical force was applied, the magnitudes of stress of the various dowel designs were similar; however, when loaded horizontally, the short dowel produced the greatest dentinal stress concentration, and the tapered dowel showed the greatest stress concentration within the cement layer. Greater deflections and higher stresses were generated with horizontal loading. CONCLUSION: The dowel and core provided little reinforcement to the remaining tooth. The direction of the functional load had a greater effect than dowel design on maximum stress and displacement. Parallel-sided dowel and cores with a length of 12 mm distributed the stress widely in the restoration and dentin, resulting in the smallest stresses. PMID- 11404757 TI - 2000 Judson C. Hickey Scientific Writing Award. Effect of impression materials on hybridized dentin. AB - STATEMENT OF PROBLEM: Tooth sensitivity after crown preparation is not an uncommon complication. For dentin bonding agents to be effective in preventing postoperative sensitivity, they must remain intact throughout the fixed prosthodontic procedures. PURPOSE: This study evaluated, by analyzing the change in dentin permeability, whether a dentin bonding agent was removed from the surface of prepared teeth in the process of making an impression. MATERIAL AND METHODS: Eighty extracted human molars were cut horizontally apical to the cemento-enamel junction and horizontally through the middle of the clinical crown. The specimens were prepared to measure dentin permeability (fluid flow rate, Lp). The specimens (n = 40) were randomly assigned to either the primer only group (PO) or the primer + adhesive group (PA). Five different impression materials were tested: vinyl polysiloxane, polyether, polysulfide, irreversible hydrocolloid, and reversible hydrocolloid. Dentinal permeability was measured after smear layer removal (control, etched Lp = ELp), after coating the dentin with either PO or PA (coated Lp = CLp), and again after making an impression (impression Lp = ILp). A 1-way ANOVA of differences, followed by a Student-Newman Keuls test (alpha=.05), was performed to compare the ELp, CLp, and ILp values. RESULTS: A statistically significant difference was found between the CLp and ELp for all 5 impression materials in both groups (PO and PA). No difference was found between the CLp and ILp values for any of the impression materials. No significant differences were found between any of the 5 impression materials or either of the dentin surface treatments (PO or PA) when measuring the CLp and ILp. CONCLUSION: The dentin bonding agent had a significant effect in decreasing the Lp of the treated specimens. None of the impression materials significantly affected the measured CLp. There was no significant difference between the PO or PA (CLp) values. PMID- 11404758 TI - Nondestructive, in vitro quantification of crown margins. AB - STATEMENT OF PROBLEM: It is important that artificial crowns fit the prepared tooth accurately, as marginal deficiencies are predisposed to plaque accumulation and lead to increased risk of periodontal disease. Various methods of evaluation for marginal fit are described in the literature, but most approaches are limited by destructive methods of assessment and/or small points of measurement. PURPOSE: This study compared, in vitro, the marginal fit of 4 types of complete crowns on human premolar teeth with the use of nondestructive profilometry. This method determined whether fit was influenced by type of crown or surface morphology of the tooth, namely, grooved or ungrooved surfaces. MATERIAL AND METHODS: Four groups of specimens were prepared for complete crowns: group BA, bonding alloy with chamfer finish line; group G, gold alloy with chamfer finish line; group PC, porcelain with a chamfer finish line; and group PS, porcelain with a shoulder finish line. Two profiles of grooved mesial and ungrooved distal surfaces of the teeth were performed: (1) teeth prepared for each type of crown and (2) teeth with crowns seated but not cemented. Marginal fit (absolute marginal discrepancy) from the finish line edge of the tooth preparations to crown edges (CE) and leading edges (LE) of crowns were measured. RESULTS: A 2-way analysis of variance for crown type and tooth surface morphology revealed significant differences between crown types for all measurement parameters, except vertical LE. The effect of surface morphology was not significant, except for vertical LE (P<.05). For all parameters, except vertical LE, the ranking of marginal fit discrepancies from greatest to least was as follows: group PC, G, BA, and PS. For vertical LE distances, the ranking was PS, BA, G, and PC (P<.05). CONCLUSION: Profilometry was used as a nondestructive, accurate method of evaluating the absolute marginal fit of different types of crowns. Marginal fits varied continuously around the circumference of each crown and made clinical assessment of fit accuracy subjective and arduous. PMID- 11404759 TI - Application of finite element analysis in implant dentistry: a review of the literature. AB - Finite element analysis (FEA) has been used extensively to predict the biomechanical performance of various dental implant designs as well as the effect of clinical factors on implant success. By understanding the basic theory, method, application, and limitations of FEA in implant dentistry, the clinician will be better equipped to interpret results of FEA studies and extrapolate these results to clinical situations. This article reviews the current status of FEA applications in implant dentistry and discusses findings from FEA studies in relation to the bone-implant interface, the implant-prosthesis connection, and multiple-implant prostheses. PMID- 11404760 TI - Dynamic fatigue properties of the dental implant-abutment interface: joint opening in wide-diameter versus standard-diameter hex-type implants. AB - STATEMENT OF PROBLEM: The clinical long-term success of single-tooth implant restorations depends, in part, on a stable connection between the prosthetic restoration and the implant body. PURPOSE: The purpose of this experiment was to investigate the fatigue life of UCLA-style abutment screws in wide-diameter versus conventionally sized dental implant restorations. MATERIAL AND METHODS: Five 3.75 x 15-mm and five 6.0 x 15-mm hexed dental implants were used. Ten frameworks were fabricated, 5 with a single UCLA-style, 3.75-mm hexed gold alloy cylinder, and 5 with a single UCLA-style, 6.0-mm hexed gold alloy cylinder. To simulate a common laboratory procedure, 2 abutment interfaces were relieved with a one-quarter round bur for both diameters. The 3.75-mm implant used a Gold-Tite central abutment screw torqued to 32 Ncm, and the 6.0-mm implant used a titanium central abutment screw torqued to 25 Ncm. Frameworks were dynamically loaded ( approximately 10 Hz) with a 120 +/- 10-N, 4-mm off-axis force. Liquid metal strain gauges were used to measure joint opening. Measurements were made at intervals of 10(3), 10(4), 10(5), and 5x10(5) cycles. Gauge output data were converted to displacement with a conversion factor determined by calibration. Linear regression analysis then was performed. RESULTS: Two observations were made in this study. Two of three 3.75-mm nonadjusted specimens and all three 6.0 mm nonadjusted specimens maintained joint closure (range of opening 0-20 microm) while measured under dynamic loading. The median joint opening at 5x10(5) cycles for 3.75-mm nonadjusted specimens was 14 +/- 7 microm; for 6.0-mm specimens, it was 11 +/- 10 microm. Both 3.75-mm adjusted specimens and 1 nonadjusted specimen failed to maintain joint closure (excess joint opening >50 microm). One of the 3.75-mm adjusted specimens had abutment screw fracture. One of two 6.0-mm adjusted specimens failed to maintain joint closure because of screw fracture. CONCLUSION: The dental implant-abutment interface of 3.75-mm and 6.0-mm externally hexed implants experienced similar joint opening after periods of dynamic loading. Laboratory adjustment of the interface significantly decreased the service life of the abutment screw joint. PMID- 11404761 TI - Extraoral maxillofacial prosthetic rehabilitation at the M. D. Anderson Cancer Center: a survey of patient attitudes and opinions. AB - STATEMENT OF PROBLEM: Measures of satisfaction after extraoral maxillofacial prosthetic rehabilitation have been sparsely reported. PURPOSE: This article reviews the care-related opinions of patients who were provided extraoral prostheses at the M. D. Anderson Cancer Center over the course of 10 years. MATERIAL AND METHODS: A questionnaire was delivered to 263 extraoral prosthetic patients to elucidate their degree of satisfaction with several parameters, including prosthetic use, care, quality, durability, longevity, and cost. In addition, issues relating to self-image, socialization frequency, and income earning ability before and after rehabilitation were surveyed. RESULTS: The views of 76 respondents demonstrated a general satisfaction with their prostheses. A majority believed that their prostheses fit comfortably, and most were satisfied with cosmesis. In addition, a preponderance of respondents reported no substantial alteration in social activity after rehabilitation. Although the number of respondents relying on their own employment fell by more than 50% after rehabilitation, nearly all reported a relatively static income before and after rehabilitation. CONCLUSION: The survey revealed a high degree of patient satisfaction with extraoral maxillofacial prostheses. Nevertheless, areas of potential improvement were not lost on the survey's population. The patients desired prostheses that last longer and have improved color stability. An interest in improved retentive mechanisms was mentioned by several patients, and only about half of the respondents perceived prosthetic cost to be completely reasonable. PMID- 11404762 TI - Accelerated color change in a maxillofacial elastomer with and without pigmentation. AB - STATEMENT OF PROBLEM: Maxillofacial prostheses require frequent replacement because the elastomer and its color additives undergo changes. PURPOSE: This study attempted to determine whether predictable color changes occur when 3 pigments are individually incorporated into a specific silicone elastomer. MATERIAL AND METHODS: The materials included an RTV elastomer; 1 natural inorganic pigment, burnt sienna; and 2 synthetic organic pigments, Hansa yellow and alizarin red. Eight test groups of 10 polymerized specimens were established. Groups 1 and 2, acting as the control, involved only the elastomer. Groups 3 and 4 were composed of elastomer and burnt sienna. Groups 5 and 6 consisted of elastomer and Hansa yellow. Groups 7 and 8 comprised elastomer and alizarin red. Odd-numbered groups were assigned to a test site in Miami, Fla., whereas the even numbered groups went to Phoenix, Ariz. Specimens weathered in Miami and Phoenix received sunlight exposures of 1305.7 MJ/m2 and 1310.2 MJ/m2, respectively, over time. Before and after weathering, the L* a* b* color parameter (DeltaE*) of each specimen was determined spectrophotometrically. RESULTS: Mean color changes that occurred in Arizona were larger than those produced in Florida. Specifically, these differences ranged from 0.4 (alizarin red groups) to 2.36 units for the 2 unpigmented control groups. Other differences showed significance for the unpigmented (P=.001), burnt sienna (P=.006), and Hansa yellow groups (P=.001). CONCLUSION: Outdoor weathering tests in which documented ASTM methods were used provided a valid baseline for future research on color changes in maxillofacial prostheses. PMID- 11404763 TI - Using the neutral zone to obtain maxillomandibular relationship records for complete denture patients. AB - A technique for obtaining maxillomandibular registration for complete denture patients is presented. The maxillary rim is formed with the use of conventional techniques. The mandibular rim is made from modeling plastic impression compound on a record base formed by the patient into the neutral zone. The mandibular rim then is reheated, and the patient determines the occlusal vertical dimension by swallowing. An imprint of the maxillary rim is made on the mandibular rim at the occlusal vertical dimension. The posterior extent of the mandibular rim is relieved 1 mm. Orientation notches are placed in both rims, and centric relation is recorded with a fast-setting vinyl polysiloxane material. PMID- 11404764 TI - The double-inlay technique: a new concept and improvement in design. AB - Gold and ceramic have long been used in prosthetic dentistry. In the posterior region, it is possible to use both materials with the double-inlay technique to add the strength of metal to the esthetics of ceramic. The problem, however, remains the volume of tooth reduction required to avoid ceramic fracture. This article describes a modification of the double-inlay technique that makes use of a pin system. This modified technique permits good retention while avoiding excessive tooth reduction. PMID- 11404765 TI - A helpful technique for removing dowel core pattern locked within a tooth. PMID- 11404766 TI - A simple, predictable method for repositioning a complete denture for soft-lining procedures. PMID- 11404767 TI - [Crystal structures of heme binding protein 23 and xanthine dehydrogenase]. PMID- 11404768 TI - [Cloning of group A streptococcal pyrogenic exotoxin-B gene and its recombinant protein expression in culture supernatant]. AB - Streptococcal pyrogenic exotoxin B, a conserved cysteine protease (SPE B/SCP) released by group A Streptococcus (GAS) strains, is considered to be an important virulence factor of this bacterium. This paper reports the cloning of gene encoding SPE B/SCP. For production of recombinant SPE B/SCP (rSPE B/SCP), the primers specific for the SPE B/SCP gene (spe b) were designed based on its nucleotide sequence. Polymerase chain reaction (PCR) was performed with the genomic DNA of GAS strain NZ131 as a template. The amplified PCR products were purified and cloned into the pBluescript II SK(+) plasmid vector. The vector was transformed into Escherichia coli (E. coli) JM109. The rSPE B/SCP and its recombinant proenzyme (rzym) were secreted in the culture supernate of the transformant. The rSPE B/SCP was purified from the supernatant by sequential chromatography on DEAE-Sepharose, matrix gel Red A and Sephadex G-50 columns. The purified rzym and rSPE B/SCP, respectively, gave a single band with a molecular weight approximately 40 kDa and 27 kDa on SDS-polyacrylamide gel electrophoresis, and reacted with anti-SPE B/SCP antibodies in Western Blot analysis. This is the first report in which rSPE B/SCP was obtained from the culture supernate of the transformant. PMID- 11404769 TI - A suspension procedure using the extensor carpi ulnaris tendon for distal radioulnar joint disorders. AB - Recently the Sauve-Kapandji (S-K) procedure has become popular for the treatment of various distal radioulnar joint (DRUJ) disorders. However, some complications, especially pain over the proximal stump of the ulna due to instability of the ulna have been reported in more recent follow-up studies. To prevent the occurrence of this pain, we devised a modified S-K procedure, which we called the suspension procedure, in which the extensor carpi ulnaris (ECU) tendon was used to suspend the proximal ulnar stump. We report here the surgical technique and compare clinical and radiographic results between the suspension procedure and the S-K procedure alone. We performed the S-K procedure alone on 8 patients (original group) and the suspension procedure on 5 (suspension group). Clinical results were assessed according to the clinical evaluation scoring system described by Inoue. Radiographic evaluations included the radio-ulnar distance, the gap of the ulna, and the distance between the articular surface of the wrist and the proximal ulnar stump. In the original group, 4 patients were rated as excellent, 2 as good and 2 as fair, whereas in the suspension group, 3 were rated as excellent, 2 as good and none as fair. In regard to radiographic evaluations, there were no significant differences in any of the 3 parameters between the 2 group. This suspension procedure had an advantage over the S-K procedure alone, especially in preventing the occurrence of stump pain. As there was no significant difference in radiographic findings between the two procedures regarding the site of osteotomy, the amount of bone resection, and radio-ulnar distance, stump pain may be attributed to dynamic instability rather than to static instability. PMID- 11404770 TI - Activation of the adenosine triphosphate sensitive mitochondrial potassium channel is involved in the cardioprotective effect of isoflurane. AB - The adenosine triphosphate-dependent potassium (K(ATP)) channel has been proposed to play an important role in the cardioprotective effect of isoflurane (ISO). However, the question of whether the K(ATP) channel, sarcolemmal or mitochondrial is the main contributor to the effect has not been clarified. The major aim of the present study was to determine whether or not the mitochondrial potassium channel was a site of action for ISO. Whether there was an acute "memory phase", in which drugs were not detected in the tissues, but the protective effect still remained in the ischemic preconditioning (IP) -like effect of ISO was also investigated. Dangling participle isolated rat hearts, a 20-min normothermic nonperfused phase was maintained to produce a global ischemia. Under these ischemic conditions, the effects of ISO, sodium 5-hydroxydecanoate (5HD: a selective mitochondrial K(ATP) channel antagonist), and ISO combined with 5HD on cardiac performance were examined. To all these four groups, (non-treated group, ISO group, 5HD group and ISO plus 5HD group, n=6 each) drugs were given for 30 min. After 10 min of drug-free perfusion (pre-ischemia restabilization period), 20 min of ischemia followed. Then the cardiac performance and the creatine kinase (CK) release during the reperfusion period were tested. In the non treated group and 5HD group, cardiac performance was stable during the treated period and pre ischemia the restabilization period. In the ISO group and ISO plus 5HD group, heart rate (HR), left ventricular (LV) systolic pressure, and LV maximum rate of development of tension (dP/dtMax) during the drug-treated period became gradually and linearly worse. However, these values were the same as in the non-treated group and 5HD group at the end of the pre-ischemia restabilization period. So 5HD itself had no hemodynamic effect; nor did it have any influence on the actions of ISO. At the end of the pre-ischemia restabilization period, the significant hemodynamic differences among the groups diminished and ISO was not detected in the solution. In the post-reperfusion period, except for the ISO group, (non treated group, 5HD group and ISO plus 5HD group) cardiac performances were drastically decreased. ISO significantly ameliorated the dysfunction of cardiac output, LV systolic pressure and LV+dP/dtMax. The CK level in the coronary effluent during reperfusion was also significantly reduced by ISO. 5HD completely inhibited these cardiac effects of ISO. Activation of the adenosine triphosphate sensitive mitochondrial potassium channel is involved in the cardioprotective effect of ISO, and the action of this agent has an acute"memory phase" like ischemic preconditioning. PMID- 11404771 TI - [A clinical study of the usefulness of assessing dyspnea in healthy elderly subjects]. AB - Dyspnea is a major clinical symptom of various respiratory diseases. However, no objective assessment of dyspnea on exertion (DOE) in elderly subjects has been established yet. Furthermore, the factors which may influence DOE in healthy elderly subjects have not yet been precisely elucidated. An oxygen cost diagram (OCD), which was originally developed by McGavin (1978), is one of the methods of assessing dyspnea on exertion in a semi-quantitative way, although it is still uncertain which factor (s) might influence the changes in OCD values. The present study was, therefore, undertaken to study; 1) whether OCD values are useful for the assessment of DOE in elderly subjects, and 2) the possible factors (s) which might contribute to changes in OCD values in these patients. The total number of subjects which were enrolled in the present study was 818, consisting of 355 men and 463 women, whose mean age was 76.4 years old, was studied. Spirometry, arterial blood gases and OCD values were measured on the same day. The OCD value and FEV(1.0) declined linearly with advanced aging. It was found that the factors which significantly reduce OCD values were as follows: aging, vital capacity, FEV(1.0), and maximal voluntary ventilation (MVV). The odd ratio which contributes to changes in OCD values was calculated. It appeared that there was a gender difference: when the odd ratio of OCD values of less than 70 was taken as 1 in the men, the odd ratio in women was calculated as 1.42. The odd ratio increased with advancing age; when the value in the 65~69 year-old group was 1, the odd ratios in the 85~89 year-old and 90~94 year-old groups were in approximately 6 and 8, respectively. Similarly, the odd ratio increased parallel with reduction in MVV. From these results, we conclude that the OCD value is reliable, simple and the best method of evaluating dyspnea in elderly subjects semi-quantitatively, and both the minute ventilatory volume and age are closely related with changes in OCD values. PMID- 11404772 TI - [Expression and intracytoplasmic signal transduction pathway of fibroblast growth factor (FGF)-10 in human cervical cancer cell lines]. AB - Fibroblast growth factor (FGF) -10 is a new member of the FGF family initially reported in Japan. It is mainly synthesized by mesenchymal cells and acts on epithelial cells in a paracrine manner. FGF-10 actions are dependent on their binding to the iiib form of FGF receptor 2 (FGFR2) iiib, also known as keratinocyte growth factor receptor (KGFR). FGF-10 has high amino acid homology to keratinocyte growth factor (KGF) and plays an important role in fetal limb and lung development and skin wound healing. In the present study, the expression of FGF-10 and FGFR2 iiib messenger RNA (mRNA) in two different human uterine cervical cancer cell lines (CaSki and ME-180) were examined. Both CaSki and ME 180 cells expressed FGFR2 iiib mRNA, while only CaSki cells expressed FGF-10 mRNA and protein. Recombinant FGF-10 (1 ng/ml) increased the growth rate of ME-180 cells and also enhanced mitogen-activated protein kinase (MAPK) phosphorylation of the cells. These data indicate that FGF-10 may directly promote the growth of squamous cell cancer in the uterine cervix via the MAPK pathway. PMID- 11404773 TI - Acute cholecystitis caused by a cholesterol polyp. AB - A 39-year-old man hospitalized with upper abdominal pain had been found to have a 3mm polyp in the body of the gallbladder 3 years previously. Laboratory tests on admission showed mild liver dysfunction. Ultrasonography depicted a dilated gallbladder with increased wall thickness; the polyp could no longer be seen. Computed tomography with drip infusion cholangiography again showed a dilated gallbladder, and also stenosis of the distal cystic duct. The resected specimen obtained by laparoscopic cholecystectomy showed disappearance of the polyp from the body of the gallbladder. A cholesterol stone was incarcerated in the cystic duct, representing an impacted detached cholesterol polyp causing acute cholecystitis. Spontaneous detachment of a cholesterol polyp from the gallbladder mucosa, then, can result in acute cholecystitis. PMID- 11404774 TI - A clinical evaluation of lymphangioma of the large intestine: a case presentation of lymphangioma of the descending colon and a review of 279 Japanese cases. AB - With the development and widespread use of colonoscopy, lymphangioma of the large intestine has recently been reported frequently. This paper presents some findings from a review of 279 cases of this disease in Japan, including a typical case that we encountered. A 69-year-old female was diagnosed as having lymphangioma of the descending colon based on the findings of a barium enema and a colonoscopy, and the lesion was successfully removed by an endoscopic resection. In the published reports, the etiology of this disease is not clear yet but the age at onset range shows a tendency toward a higher incidence in comparatively older patients and the male-to-female ratio indicates a higher incidence in males. If there is no complication, endoscopic treatment seems to be the preferable procedure for this disease. PMID- 11404775 TI - Hemorrhagic gastric carcinoma in an acromegalic patient. AB - A rare case of hemorrhagic gastric carcinoma in an acromegalic patient is reported. A 79-year-old Japanese man was referred to our hospital with diagnoses of upper gastrointestinal hemorrhage and angina pectoris. This patient showed typical clinical features of acromegaly, with increased serum growth hormone (GH) and insulin-like growth factor I (IGF-I) level. A high titer of serum anti Helicobacter pylori (H. pylori) IgG was also observed. After percutaneous transluminal coronary angioplasty treatment for stenosis of the right coronary artery, the patient underwent distal gastrectomy. Gastric cancer was Type 2 macroscopically and was diagnosed histologically as a papillary and well to moderately differentiated tubular adenocarcinoma. Reverse transcription polymerase chain reaction analysis estimated that the amount of IGF-I receptor mRNA expression in the gastric cancer tissue was 1.6 times higher than that in the adjacent atrophic mucosa, whereas the amount of IGF-I mRNA expression in the cancer tissue was only half that in the atrophic mucosa. Both the stimulatory effects of GH and/or IGF-I on cell proliferation and H. pylori infection in gastric tumorigenesis may have been responsible for the development and growth of gastric carcinoma in this patient. PMID- 11404776 TI - An elderly patient with gastric carcinoma developing multiple metastasis in skeletal muscle. AB - We present a 70-year-old man with gastric carcinoma developing multiple metastasis in skeletal muscle. He had a right supraclavicular lymph node swelling. Brain, chest and abdomen CT scans revealed metastatic lesions in the brain, lung, liver and bilateral adrenal glands. Further, CT showed a ring enhanced soft-tissue mass in the left lumbar muscle. Needle aspirate of the mass in both the left lumbar muscle and the right enlarged supraclavicular lymph node revealed cells suggestive of poorly differentiated adenocarcinoma. Upper gastrointestinal endoscopic evaluation demonstrated an advanced gastric carcinoma. Two months after admission, the tumor in the left lumbar muscle had grown and some new lesions in the left iliopsoas muscles appeared. Intramuscular metastasis from gastric carcinoma is an extremely rare phenomenon. PMID- 11404777 TI - [Nephron-sparing endoscopic treatment using a Ho: YAG laser of three patients with upper urinary tract urothelial tumors]. PMID- 11404783 TI - Response of the brain to enrichment. AB - Before 1960, the brain was considered by scientists to be immutable, subject only to genetic control. In the early sixties, however, investigators were seriously speculating that environmental influences might be capable of altering brain structure. By 1964, two research laboratories proved that the morphology and chemistry or physiology of the brain could be experientially altered (Bennett et al. 1964, Hubel and Wiesel 1965). Since then, the capacity of the brain to respond to environmental input, specifically "enrichment," has become an accepted fact among neuroscientists, educators and others. In fact, the demonstration that environmental enrichment can modify structural components of the rat brain at any age altered prevailing presumptions about the brain's plasticity (Diamond et al. 1964, Diamond 1988). The cerebral cortex, the area associated with higher cognitive processing, is more receptive than other parts of the brain to environmental enrichment. The message is clear: Although the brain possesses a relatively constant macro structural organization, the ever-changing cerebral cortex, with its complex microarchitecture of unknown potential, is powerfully shaped by experiences before birth, during youth and, in fact, throughout life. It is essential to note that enrichment effects on the brain have consequences on behavior. Parents, educators, policy makers, and individuals can all benefit from such knowledge. PMID- 11404784 TI - Functional role of a glycolipid in directional movements of neurons. AB - Migration of neurons from their site of origin to their final destination is a critical and universal step in the formation of the complex structure of the nervous system. The migratory process is thought to be governed in part by genetically and epigenetically defined sequences of signals which are interpreted by migrating cells. The molecular mechanisms that underlie neuronal migration have been the subject of intense investigation. As in other developmental processes, many molecules must participate in neuronal migration. Some molecules, such as cell adhesion molecules and motor proteins, may contribute to discrete steps in the migration act; others, like extracellular signaling molecules, may regulate the activation and/or termination of the migration program. In this article we review findings from our group that demonstrate the functional role(s) of a specific glycolipid in neuronal migration and neurite outgrowth in the developing and adult nervous system. PMID- 11404785 TI - Watery and dark axons in Wallerian degeneration of the opossum's optic nerve: different patterns of cytoskeletal breakdown? AB - In this paper we report a qualitative morphological analysis of Wallerian degeneration in a marsupial. Right optic nerves of opossums Didelphis marsupialis were crushed with a fine forceps and after 24, 48, 72, 96 and 168 hours the animals were anaesthetized and perfused with fixative. The optic nerves were immersed in fixative and processed for routine transmission electron microscopy. Among the early alterations typical of axonal degeneration, we observed nerve fibers with focal degeneration of the axoplasmic cytoskeleton, watery degeneration and dark degeneration, the latter being prevalent at 168 hours after crush. Our results point to a gradual disintegration of the axoplasmic cytoskeleton, opposed to the previous view of an "all-or-nothing" process (Griffin et al 1995). We also report that, due to an unknown mechanism, fibers show either a dark or watery pattern of axonal degeneration, as observed in axon profiles. We also observed fibers undergoing early myelin breakdown in the absence of axonal alterations. PMID- 11404788 TI - Environmental risk increase due to heavy metal contamination caused by a copper mining activity in Southern Brazil. AB - The Camaqua Copper Mines (CCM) were the main sulphide deposit in Southern Brazil and have been in operation from last century to 1996. To evaluate water contamination and environmental risk increase by heavy metals from mining operations, two points on the Joao Dias Creek were sampled (Station 1, background area and Station 2, contaminated area). Mining activity increased the natural weakly heavy metal fluxes by approximately 5424 kg. (approximately 60%) of the total metal flux, 1542 kg. (approximately 49%) of dissolved and 3881 kg (approximately 66%) of particulate metal flux. Total metal flux of anthropic origin was mostly due to Fe followed by Cu > Zn > Mn whereas Cd, As and Pb fluxes were negligible. The potential human health hazards and risk assessment related to daily intake of water from Joao Dias Creek are mostly due to Mn and should be of concern for the contaminated area. The ingestion of water from station 2 represents incremental risks of 130% and 59% respectively, considering the non carcinogenic and the carcinogenic effects. The real increase of human health hazards may be greater than those related to the total concentrations since Mn and As dissolved concentrations were 5.5 and 2.0 higher than acceptable, respectively. PMID- 11404790 TI - Role of platelet-activating factor in reproduction: sperm function. AB - Since its discovery nearly thirty years ago, platelet-activating factor has emerged as one of the more important lipid mediators known. Platelet-activating factor (PAF; 1-O-alkyl-2-O-acetyl-sn-glycero-3-phosphorylcholine) exists endogenously as a mixture of molecular species with structural variants of the alkyl moiety. PAF is a novel potent signaling phospholipid that has unique pleiotropic biological properties in addition to platelet activation. PAF also plays a significant role in reproduction. PAF content in squirrel monkey sperm is significantly higher during the breeding season than the non-breeding season. PAF content in human sperm has a positive correlation with seminal parameters and pregnancy outcomes. High-fertility boars have significantly more PAF in their sperm than low-fertility boars. The enzymes (lyso-PAF-acetyltransferase and PAF acetylhydrolase) necessary for PAF activation and deactivation are present in sperm. PAF-acetylhydrolase may act as a "decapacitation factor". Removal of this enzyme during capacitation may promote PAF synthesis increasing motility and fertilization. PAF also plays a significant role in the fertilization process, enhancing the fertilization rates of oocytes. Enhanced embryo development has also been reported in oocytes fertilized with PAF-treated sperm. PAF antagonists inhibit sperm motility, acrosome reaction, and fertilization, thus suggesting the presence of receptors for PAF. The PAF-receptor is present on sperm, with altered transcript levels and distribution patterns on abnormal cells. Whereas the exact mechanism of PAF in sperm function and reproduction is uncertain, its importance in normal fertility is substantial. The reproductive significance of PAF activity in sperm and fertility plus the role of PAF in the establishment of pregnancy requires further study. PMID- 11404791 TI - Vas deferens, a site of male contraception: an overview. AB - The vas deferens is a site which can be exploited for male contraception without undue side effects. The only effective technique available for male contraception is vasectomy, being practiced world wide, despite that it is a permanent surgical procedure and its successful reversal is not assured. Although no-scalpel vasectomy minimizes surgical procedures, the fate of its reversal is akin to that of vasectomy. Several occlusive and non-occlusive vasal procedures which claim to be reversible without surgical intervention, possess more disadvantages than advantages. Vas occlusion with plug, 'Shug' or medical grade silicone rubber, although claimed to produce reversible azoospermia without affecting spermatogenesis, requires skilled microsurgery for their implantation and later removal. RISUG, a non-sclerotic polymer styrene maleic anhydride (SMA), could be more advantageous than vasectomy and other vas occlusive procedures in that it could be a totally non-invasive procedure by "no-scalpel injection" and "non invasive reversal". It is claimed to offer long-term contraception without adverse side effects and also to be possible as a male spacing method by repeated vas occlusion and non-invasive reversal. The drug is currently under multicentre Phase III clinical trial. PMID- 11404792 TI - L-arginine, the substrate of nitric oxide synthase, inhibits fertility of male rats. AB - AIM: To examine the effect of L-arginine, the substrate of nitric oxide (NO) synthase, on reproductive function of male rats. METHODS: Male rats were gavaged with either L-arginine (100 or 200 mg x kg(-1) x d(-1)), D-arginine (200 mg x kg( 1) x d(-1)) or vehicle (0.9% NaCl) for seven consecutive days. Their sexual behaviour and fertility were evaluated using receptive females. RESULTS: L arginine (200 mg/kg) had no significant effect on sexual competence (in terms of sexual arousal, libido, sexual vigour and sexual performance). In mating experiments, the higher dose of L-arginine effectively and reversibly inhibited fertility, whilst the lower dose and the inactive stereoisomer D-arginine had no significant effect. The antifertility effect caused by L-arginine was due to a profound elevation in the preimplantation loss mediated possibly by impairment in epididymal sperm maturation, hyperactivated sperm motility and sperm capacitation. CONCLUSION: Elevated NO production may be detrimental to male fertility. PMID- 11404793 TI - Castration-induced expression of caspase-1 in epithelia of accessory sex organs in male rats. AB - AIM: As an attempt to clarify the molecular basis of castration-induced apoptosis, this study was undertaken to demonstrate the expression of caspase-1 in male accessory sex organs of rats. METHODS AND RESULTS: cDNA of rat caspase-1 was cloned by reverse transcription-polymerase chain reaction from the ventral prostates. The open reading frame predicts 402 amino acids, which shows more than 91% and 63% identity to those of mouse and human, respectively. Northern analyses demonstrated the presence of castration-induced up-regulation of the 1.6 kb transcript in the ventral prostate and the seminal vesicles. Finally, the authors demonstrated the caspase-1 transcripts in the epithelia of these tissues by in situ hybridization analyses. CONCLUSION: Castration induces the expression of caspase-1 transcripts in the epithelia of ventral prostate and seminal vesicle. These observations suggest a possible role of caspase-1 in apoptosis in male accessory sex organs. PMID- 11404794 TI - Evaluation of effects of 1,3-dinitrobenzene on sperm motility of hamster using computer assisted semen analysis (CASA). AB - AIM: To evaluate the effects of 1,3-dinitrobenznene (mDNB) on sperm motility of hamster and to correlate the results with the fertility. METHODS: Adult male hamsters were gavaged with one of the 3 dose regimes of mDNB (1.5 mg daily for 4 weeks, 1.5 mg one day a week for 4 weeks and 1.0 mg 3 days a week for 4 weeks). Computer assisted semen analysis (CASA) was used to analyse the sperm motility parameters, curvilinear velocity (VCL) and straight line velocity (VSL) of sperm in distal corpus epididymides and distal cauda epididymides. In vitro fertilisation was carried out only for 1.5 mg mDNB daily group to determine the sperm fertilising capacity. RESULTS: There was a significant reduction in sperm velocity parameters at weeks 3 and 4 after treatment, which was correlated with a decline in sperm fertility. CONCLUSION: Sperm velocity parameters may be used to determine the effect of a toxic insult on the sperm function. PMID- 11404795 TI - Effect of intermittent treatment with tamoxifen on reproduction in male rats. AB - AIM: To identify the antifertility effect of intermittent oral administration of tamoxifen in male rat. METHODS: Tamoxifen was administered orally at a dose of 0.4 mg x kg(-1) x d(-1) with an intermittent regime for 120 days. Treated and control rats were mated with cycling female rats on days 60, 90 and 120 of treatment. The mated males were sacrificed and the weights of reproductive organs were recorded, and the serum levels of LH, FSH, testosterone and estradiol estimated by radioimmunoassay. In the female rats, the numbers of implantation sites, corpora lutea, and numbers of normal and resorbed foetuses were recorded on d 21 of gestation. The potency, fecundity, fertility index, litter size and post-implantation loss were then calculated. RESULTS: The fecundity of male rats was completely suppressed by tamoxifen while the potency was maintained at the control level. The fertility index was significantly decreased. No viable litters were sired. Post implantation loss, indicative of non-viable embryos, was observed but was not significantly increased above the control level. The weights of the testes, epididymides, ventral prostate and seminal vesicles were significantly reduced. The blood LH and testosterone levels were significantly decreased, but not FSH and estradiol. CONCLUSION: Intermittent oral tamoxifen administration completely suppressed the fecundity of adult male rats with reserved potency. PMID- 11404796 TI - Effect of testicular capsulotomy on lipid droplets in the seminiferous tubules of rats. AB - AIM: In order to reveal the histochemical alteration that might occur during the processes of the spermatogenic disruption induced by testicular capsulotomy, the location and alteration of lipid droplets in the seminiferous tubules were observed in the present study. METHODS: Osmium tetroxide was used to demonstrate the lipid droplets in the seminiferous tubules of capsulotomized and sham operated control testes. RESULTS: In the seminiferous tubules of the sham operated rat testes, many small lipid droplets were located close to the basement membrane of the seminiferous tubules. But for the capsulotomized testes, the lipid droplets in the seminiferous tubules had increased in size and number, with many lipid droplets migrated towards the lumen of the tubules. CONCLUSION: The results indicated that a progressive fatty degeneration occurred in the seminiferous tubules after testicular capsulotomy. PMID- 11404797 TI - Comparison between the quality and function of sperm after semen processing with two different methods. AB - AIM: To compare the recovery rate of morphologically normal and chromatin condensed spermatozoa from native semen samples using the SpermPrep filtration columns and Percoll gradient centrifugation and to determine the influence of the two processing techniques on fertilization and pregnancy rates in an IVF-ET program. METHODS: Sixteen semen samples obtained from patient's husband were included in this study. Each was divided into two aliquots. The first aliquot was processed with SpermPrep filtration columns and the second, Percoll gradient centrifugation. Smears were made before and after semen processing with both methods for the evaluation of chromatin condensation (chromomycine CMA3) as well as morphology (strict criteria) of spermatozoa. One hundred and seventy oocytes were retrieved from the patients and the oocytes from each patient were subdivided into two sets: one set was inseminated using spermatozoa processed with SpermPrep and the other inseminated after semen processing with Percoll gradient centrifugation. RESULTS: The Percoll method yielded a significantly higher percentage of chromatin condensed (90.8 +/- 6.5% vs 82.3 +/- 8.8%, P = 0.017) and morphologically normal spermatozoa (12.9 +/- 7.4% vs 6.9 +/- 4.8%, P = 0.001) in comparison to SpermPrep. Whereas, sperm count recovery rate was significantly higher after the use of SpermPrep than after the Percoll gradient centrifugation. The fertilization rate was similar between the two methods. CONCLUSION: Semen processing with Percoll should be recommended for intracytoplasmic sperm injection as the natural selection is bypassed and the SpermPrep technique could be recommended for IVF and IUI programs as the sperm concentration plays a more significant role in these procedures. PMID- 11404798 TI - Apoptosis and hormonal milieu in ductal system of normal prostate and benign prostatic hyperplasia. AB - AIM: To study the apoptotic rate (AR) and the androgen and estrogen milieu in the proximal and distal ductal systems of prostate, in order to help exploring the effects of these factors on prostatic growth and the pathogenesis of benign prostatic hypertrophy (BPH). METHODS: The proximal and distal ends of the ductal system were incised from 20 normal prostate as well as the hypertrophic prostate tissue from 20 patients with BPH. The AR was determined by the DNA end-labeling method and dihydrotestosterone (DHT) and estrodiol (E2), by radioimmunoassay. RESULTS: There was no significant difference in DHT and E2 density between the proximal and distal ends of the ductal systems in normal prostate. E2 appeared to be higher in BPH than in normal prostatic tissues, but the difference was statistically insignificant. In normal prostatic tissue, the AR was significantly higher in the distal than in the proximal ends of the ductal system (P < 0.05), while the AR of the proximal ends was significantly higher (P < 0.01) than that in the BPH tissue. No significant correlation was noted between the DHT and E2 density and the AR both in the normal prostate and BPH tissues. CONCLUSION: The paper is the first time describing a difference in AR in different regions of the ductal system of normal prostate, while the hormonal milieu is similar, indicating a functional inhomogeneity of these regions. A low AR in the proximal duct, where BPH originates, and an even lower AR in the BPH tissue, suggesting the participation of apoptosis in the BPH pathogenesis. PMID- 11404799 TI - Effect of lindane on testicular antioxidant system and steroidogenic enzymes in adult rats. AB - AIM: To find out the effect of lindane on testicular antioxidant system and testicular steroidogenesis in adult male rats. METHODS: Adult male rats were orally administered with lindane at a dose of 5.0 mg/kg body weight per day for 30 days. Twenty-four hours after the last treatment the rats were killed using anesthetic ether. Testes, epididymis, seminal vesicles and ventral prostate were removed and weighed. A 10% testicular homogenate was prepared and centrifuged at 4 degrees C. The supernatant was used for various biochemical estimations. RESULTS: The body weight and the weights of testes, epididymis, seminal vesicles and ventral prostate were reduced in lindane-treated rats. There was a significant decline in the activities of antioxidant enzymes superoxide dismutase (SOD), catalase and glutathione reductase while an increase in hydrogen peroxide (H2O2) generation was observed. The specific activities of testicular steroidogenic enzymes 3beta-hydroxysteroid dehydrogenase and 17beta hydroxysteroid dehydrogenase were decreased. The levels of DNA, RNA and protein were also decreased in lindane-treated rats. CONCLUSION: Lindane induces oxidative stress and decreases antioxidant enzymes in adult male rats. PMID- 11404800 TI - Effect of diabetes and insulin treatment on nitric oxide synthase content in rat corpus cavernosum. AB - AIM: To study the effect of diabetes mellitus and insulin treatment on rat penile nitric oxide synthase content. METHODS: Male Wistar rats were divided at random into two groups: the Control (n = 8) and the Diabetic (n = 17). Diabetes mellitus was induced by intraperitoneal injection of streptozotocin. The diabetic animals were then randomly divided into two subgroups: diabetic rats without insulin treatment (n = 7) and diabetic rats with insulin treatment (n = 10). The neuronal nitric oxide synthase (nNOS) in the penile corpus cavernosum were assayed by immumohistochemical staining with specific antibody to nNOS and the nNOS-positive nerve fibers were counted semiquantitatively under a high power microscope. RESULTS: The nNOS- positive nerve fibres in diabetic rats with treatment was higher than that in diabetic rats without treatment (P < 0.05) and lower than that in the controls (P < 0. 01). The nNOS-positive nerve fibres in diabetic rat without treatment were also lower than that in the controls (P < 0. 01). CONCLUSION: In streptozotocin-induced diabetic rats, the nNOS content in the penile corpus cavernosum was significantly decreased. Insulin treatment at the dose level employed partially restores the penile nNOS content in these rats. PMID- 11404801 TI - Effect of papaya seed extract on microenvironment of cauda epididymis. AB - AIM: To evaluate the effect of aqueous Carica papaya seed extract on microenvironment of cauda epididymis. METHODS: Adult male albino rats were intramuscularly administered with 0 (control) or 0.5 mg papaya seed extract/kg body weight for 7 days. Cauda epididymal tubular content was collected by micropuncture technique; epididymal luminal fluid and sperm pellets were separately analyzed. RESULTS: The results revealed that the extract treatment caused significant reduction, as compared with control, in total protein and sialic acid contents in both epididymal fluid and sperm pellet. As compared with control, significantly lowered acid phosphatase activity was recorded in sperm pellet but was higher in epididymal fluid after the treatment. The extract treatment also caused significant reduction in level of inorganic phosphorus in the epididymal fluid. CONCLUSION: It is concluded that the aqueous papaya seed extract alters cauda epididymal microenvironment. PMID- 11404802 TI - Management of symptomatic ureteric stumps laparoscopically. AB - AIM: To study the advantage of excision of the distal symptomatic ureteric stumps with the retroperitoneal laparoscopic approach. METHODS: Four patients who had failed to settle their symptoms with the initial conservative management were included in the study. All underwent excision of the distal symptomatic ureteric stumps with the retroperitoneal laparoscopic approach and then received prophylactic antibiotics. RESULTS: We have achieved better results than those reported in the literature in terms of operating time (mean 1 h 45 min), blood loss (< 10 mL), postoperative recovery (within 12 h) and hospital stay (< 48 h). CONCLUSION: Retroperitoneal laparoscopic excision is a safe, simple and effective method in the management of symptomatic ureteric stumps. PMID- 11404803 TI - Effect of testicular capsulotomy on secretion of testosterone and gonadotrophins in rats. PMID- 11404804 TI - Developmental memory performance: inter-task consistency and base-rate variability on the WRAML. AB - Two studies examined developmental memory test consistency and base-rate variability on the Wide Range Assessment of Memory and Learning (WRAML) using three age cohorts from the standardization sample. Study 1 examined inter-subtest correlation coefficients across the nine subtests of the WRAML and compared across three age cohorts (5, 11, 16 to 17 year olds). An age-related increase in inter-task consistency was found (mean r = .26 and .42 for the youngest and oldest age groups, respectively). However, correlation coefficients were generally in the low to moderate range (rs = .2 to .5) for all three cohorts suggesting considerable performance variability across memory subtests. Study 2 examined base-rate variability in the WRAML standardization sample using several different methods. More specifically, base-rate information is provided for the maximum discrepancy between subtests, profile strengths and weakness (i.e., discrepancies from the mean scaled score), and the prevalence of individuals within the "deficient" performance range (i.e., < = 2 SD below normative means). In addition, performance variability across the four WRAML index scores was examined by determining the prevalence rates for the maximum discrepancy (1) between index scores, and (2) from the General Memory Index compared to the other three index scores. Performance discrepancies tended to be higher among the youngest group. Again, however, considerable performance variability was observed across all three age cohorts. Implications for clinical practice are discussed. PMID- 11404805 TI - Severe developmental prosopagnosia in a child with superior intellect. AB - Developmental prosopagnosia, a lifelong inability to learn and recognize familiar faces, has rarely been reported, and there are even fewer cases that have been studied during childhood. Of the cases studied during childhood, significant "apperceptive" features to the face recognition defect have been noted. We had an opportunity to conduct extensive standard and experimental neuropsychological, psychophysiological, and neuroanatomical studies in a five-year-old child with severe developmental prosopagnosia. The subject was intellectually gifted (FSIQ = 130), but had a marked discrepancy between verbal and nonverbal abilities (VIQ = 140, PIQ = 110). Although some visual perceptual weaknesses were apparent, the subject's face recognition defect was found to cnform most closely to the "associative" type, and he did not have visual recognition deficits for any categories of nonunique entities. A novel finding was that the child's covert recognition of familiar faces based on an autonomic index was normal, suggesting that as in some adult-onset cases, the brain is capable of acquiring some information about familiar faces, even without conscious recognition. The child also had normal judgments of facial emotional expressions. Our report extends the understanding of the neuropsychological features of developmental prosopagnosia, and may help narrow the search for neuroanatomical correlates of this condition, which have yet to be identified. PMID- 11404806 TI - An evaluation of the NINCDS-ADRDA neuropsychological criteria for the assessment of Alzheimer's disease: a confirmatory factor analysis of single versus multi factor models. AB - Neuropsychological test batteries are frequently used to assess the nature and severity of cognitive deficits among patients with early Alzheimer's Disease (AD) and related disorders. The NINCDS-ADRDA criteria are among the most widely used guidelines to diagnose dementia (McKhann et al.,1984). These criteria specify eight distinct areas of neuropsychological function that should be evaluated in patients with suspected cognitive impairment. Recent studies have suggested that neuropsychological deficits observed in AD may be explained by a single general factor related to memory deficits or to executive dysfunction. In contrast, the results of other investigations have indicated that multiple qualitatively different factors underlie cognitive abilities in AD. In the present study, we used confirmatory factor analysis to examine the structure of cognitive abilities in AD and to assess the extent to which single and multiple ability factors accurately represent neuropsychological test data obtained from patients with AD. Results indicated that the NINCDS-ADRDA model fit the data better than a single factor model. However, a more parsimonious model specifying memory, verbal abilities, visuospatial skills, executive function, and higher as well as lower functional activities of daily living fit the data better than the NINCDS-ADRDA model. These results have important theoretical and practical implications for diagnostic evaluation. PMID- 11404807 TI - Associating semantic space abnormalities with formal thought disorder in schizophrenia: use of triadic comparisons. AB - Recent studies of schizophrenia have suggested that thought disorder results from abnormalities in semantic processing. In the following pilot study, the cognitive system used for organizing and associating concepts was examined using a triadic comparison task. The semantic maps of schizophrenia patients with high thought disorder (N = 5) were compared to that of schizophrenia patients with low levels of thought disorder (N = 5) and normal controls (N = 10) with multidimensional scaling analysis. At initial testing and at retest, patients with high levels of thought disorder exhibited consistently lower semantic goodness of fit scores and failed to map results of triadic comparisons along well-defined dimensions. Results suggest that thought disorder in schizophrenia is related to a disturbance in the organization of semantic networks. PMID- 11404808 TI - Stability of visual field enlargements following computer-based restitution training -- results of a follow-up. AB - In a previous randomized placebo-controlled clinical trial, we observed significant visual field enlargements induced by computer-based restitution training in patients with cerebral lesions (Kasten et al., Nature med., 4, 1998, 1083-87). Now we asked the question whether this effect is stable after training was discontinued? Here we report data of a follow-up study after a training-free interval (mean 23.5 +/- 2.3 months after end of therapy). 16 patients of the original restitution group and 6 patients of the placebo group were re-examined. On average, in high resolution computer campimetry (stimulus detection: PeriMa, form recognition: PeriForm, color perception: PeriColor) as well as in conventional automatic perimetry (TAP-2000) both groups showed no significant decline in the number of correctly detected stimuli after training was discontinued. However, cluster analysis revealed three different types of patients, who showed either increase (Type-I), decrease (Type-II) or stability (Type-III) in performance. We propose that many patients learn to use the regained visual capacities not only in the setting of a computer training but also in every day life, while other patients do not use the areas of restored vision and show a decrease of visual functions after the end of training. The Type-I group does not need continuous training, while the Type-II group may benefit from phases of refreshment exercises. PMID- 11404809 TI - Performance on paced serial addition tasks indicates an associative network for calculation. AB - Although paced serial addition (PSA) tasks are considered to be tests of general information-processing capacity, recent work suggests that performance on such tasks is influenced by arithmetic-specific variables. We designed two visual PSA experiments to determine whether the performance of normal adults would support predictions derived from the cognitive psychology of calculation. Experiment 1 showed that mixing familiar (Arabic numeral) and less familiar (Roman numeral) stimulus formats reduced scores below the averaged scores for pure Arabic and Roman lists. The Roman-Arabic order of addends was more difficult than the Arabic Roman order. Experiment 2, which involved only Arabic numerals as addends, showed that performance could be impaired by constraining the trial-to-trial variability of sums. The results of both experiments confirm the importance of arithmetic specific variables in PSA and provide support for an associative network model of calculation. In addition, the findings implicate interference from extraneous addends and responses as the performance-limiting factor. PMID- 11404810 TI - Differentiation of attention-deficit/hyperactivity disorder subtypes: application of a neuropsychological model of attention. AB - This study represents a neuropsychological attempt to differentiate subtypes of Attention Deficit/Hyperactivity Disorder (ADHD). Participants (N = 80) were grouped by gender and ADHD subtype-Predominantly Inattentive (ADHD-I) versus Combined (ADHD-C)-resulting in four age-matched groups. Statistical analysis revealed significant differences in performance on selective attention tasks for those with ADHD-I and those with ADHD-C. Relative to their counterparts without hyperactivity, participants with ADHD-C earned disproportionately lower scores on tasks associated with executive control. Both subgroups with ADHD-I and ADHD-C demonstrated significant difficulty on some tasks assessing complex mental operations relative to age-standardized normative data. Discriminant analysis revealed that a combination of five neuropsychological measures discriminated between the ADHD-I and ADHD-C subgroups with 80% accuracy. Results provide support for the notion of the Predominantly Inattentive and Combined subtype classifications as defined by the Diagnostic and Statistical Manual of Mental Disorders, 4th Edition (American Psychiatric Association, 1994). Further, findings from this study lend preliminary support for the utility of a neurophysiologically sensitive model of attention in the differentiation of these subtypes. PMID- 11404811 TI - Assessing attention: comparison of response-inhibition and traditional continuous performance tests. AB - The purpose of this study was to compare a recently developed response-inhibition task (Conners, 1995) to slow- and fast-event-rate versions of the traditional A-X Continuous Performance Test (CPT). Among 146 normal adults, results revealed significant differences between tasks in omission and commission error rates, reaction time, reaction time variability, and responses to critical signals. Effects of environmental noise and participant anxiety also differed for the three tasks. Traditional CPTs produced time-related performance decrements, but the response-inhibition task produced improvement across initial blocks of trials. The response-inhibition task may measure "executive control" rather than sustained attention, and therefore may represent functions of different brain systems. PMID- 11404812 TI - Goal setting and reaction time performance in brain-damaged patients. AB - A goal-setting approach was used to examine the way in which 62 patients with closed head injuries or cerebral vascular accidents and 47 orthopedic control patients alter their performance on a four-choice reaction time (RT) task. Both patient groups were randomly assigned to two conditions: one in which a specific and high goal was assigned and one in which a "do your best" goal was given. Statistical analysis indicated that patients with a specific and high goal responded faster than patients with a "do your best" goal. No clinical or neuropsychological variables (e.g., attention, memory) had a moderating influence on the goal setting effect. These results demonstrate that goal setting as a motivational technique is a reliable and robust technique and can enhance performance (intensity of behavior) not only in healthy participants but also in brain-damaged patients. PMID- 11404813 TI - Neuropsychological assessment of response inhibition in adults with ADHD. AB - Several theoretical models suggest that the core deficit of ADHD is a deficiency in response inhibition. While neuropsychological deficits in response inhibition are well documented in ADHD children, research on these deficits in adult ADHD populations is minimal. Twenty-five adult ADHD patients, 15 anxiety-disordered adult patients, and 30 normal adults completed three neuropsychological tests of response inhibition: the Continuous Performance Test, Posner Visual Orienting Test, and the Stop Signal Task. ADHD adults demonstrated response inhibition performance deficits when compared to both normal adults and anxiety disordered adults only on the Continuous Performance Test. A similar pattern of differences was not observed on the other two neuropsychological tests. Differing results between tasks may be due to differences in test reliability, task parameters, or the targeted area of brain functioning assessed by each test. PMID- 11404814 TI - Chinese writing function in patients with left versus right putaminal hemorrhage. AB - This study was designed to explore the Chinese writing function of patients with subcortical stroke. Two cohorts of patient subjects with either the left or the right subcortical strokes, mainly putaminal hemorrhagic strokes, and one group of normal controls participated in the study. All participants received a writing test battery including the three aspects of writing function, that is, Spontaneous Writing, Writing to Dictation, and Writing from Copy, as well as a battery of non-writing linguistic tests. Comparing with normal controls revealed that writing function change occurred in both patient groups. The deficits in the patients with the left subcortical stroke essentially included Spontaneous Writing, and Writing to Dictation. These impairments were most likely secondary to aphasic disorders. The writing problem, mainly Writing from Copy, was noted in the patients with the right subcortical stroke. This deficit, however, was independent of the core linguistic impairment. On the basis of the results, we suggest that the lesion involving white matter in the left hemisphere probably interrupts left perisylvian cortical language organization in a manner that produces problems with spontaneous writing and writing to dictation, which are language-related, associated with lesion in the dominant hemisphere. This further suggests that left or right subcortical lesions in the putamen and surrounding white matter are associated with differential effects (language vs. non-language based effects) which are similar to such differences observed with left vs. right cortical lesions. PMID- 11404815 TI - The covert orienting of visual attention following severe traumatic brain injury. AB - Attentional problems have frequently been identified following traumatic brain injuries (TBIs) using both clinical assessments and self-report measures. Unfortunately, most measures of attention do not enable us to determine the underlying basis of these attentional deficits. One exception is Posner's Covert Orienting of Attention Task (COAT), which is designed to identify some of the fundamental mental operations underlying attention. This study sought to determine whether the COAT task could identify discrete attentional deficits following TBI beyond those caused by reduced speed of information processing. Thirty five patients who had sustained a severe TBI were compared to 35 age matched controls. Results revealed that, although the reaction times of the patients with TBI were significantly slower than the controls, there were no differences between the two groups in terms of their ability to disengage, move, and engage their attention. The introduction of a secondary (language) task produced no significant difference between the two groups on the COAT task. However, there was a significant difference between the two groups on the language-based task, suggesting a deficit in auditory-verbal attention under dual task conditions. PMID- 11404816 TI - A survey of affected-sibship statistics for nonparametric linkage analysis. AB - We have compared the power of a large number of allele-sharing statistics for "nonparametric" linkage analysis with affected sibships. Our rationale was that there is an extensive literature comparing statistics for sibling pairs but that there has not been much guidance on how to choose statistics for studies that include sibships of various sizes. We concentrated on statistics that can be described as assigning scores to each identity-by-descent-sharing configuration that a pedigree might take on (Whittemore and Halpern 1994). We considered sibships of sizes two through five, 27 different genetic models, and varying recombination fractions between the marker and the trait locus. We tried to identify statistics whose power was robust over a wide variety of models. We found that the statistic that is probably used most often in such studies-S(all) performs quite well, although it is not necessarily the best. We also found several other statistics (such as the R criterion, S(robdom), and the Sobel-and Lange statistic C) that perform well in most situations, a few (such as S(-#geno) and the Feingold-and-Siegmund version of S(pairs)) that have high power only in very special situations, and a few (such as S(-#geno), the N criterion, and the Sobel-and-Lange statistic B) that seem to have low power for the majority of the trait models. For the most part, the same statistics performed well for all sibship sizes. We also used our results to give some suggestions regarding how to weight sibships of different sizes, in forming an overall statistic. PMID- 11404817 TI - A genome screen of families with multiple cases of prostate cancer: evidence of genetic heterogeneity. AB - We conducted a genomewide screen for prostate cancer-susceptibility genes on the basis of data from 98 families from the United States and Canada that had three or more verified diagnoses of prostate cancer among first- and second-degree relatives. We found a statistically significant excess of markers for which affected relatives exhibited modest amounts of excess allele-sharing; however, no single chromosomal region contained markers with excess allele-sharing of sufficient magnitude to indicate unequivocal evidence of linkage. Positive linkage signals of nominal statistical significance were found in two regions (5p q and 12p) that have been identified as weakly positive in other data sets and in region 19p, which has not been identified previously. All these signals were considerably stronger for analyses restricted to families with mean age at onset below the median than for analyses of families with mean age at onset above the median. The data provided little support for any of the putative prostate cancer susceptibility genes identified in other linkage studies. PMID- 11404818 TI - Are rare variants responsible for susceptibility to complex diseases? AB - Little is known about the nature of genetic variation underlying complex diseases in humans. One popular view proposes that mapping efforts should focus on identification of susceptibility mutations that are relatively old and at high frequency. It is generally assumed-at least for modeling purposes-that selection against complex disease mutations is so weak that it can be ignored. In this article, I propose an explicit model for the evolution of complex disease loci, incorporating mutation, random genetic drift, and the possibility of purifying selection against susceptibility mutations. I show that, for the most plausible range of mutation rates, neutral susceptibility alleles are unlikely to be at intermediate frequencies and contribute little to the overall genetic variance for the disease. Instead, it seems likely that the bulk of genetic variance underlying diseases is due to loci where susceptibility mutations are mildly deleterious and where there is a high overall mutation rate to the susceptible class. At such loci, the total frequency of susceptibility mutations may be quite high, but there is likely to be extensive allelic heterogeneity at many of these loci. I discuss some practical implications of these results for gene mapping efforts. PMID- 11404819 TI - Multifactor-dimensionality reduction reveals high-order interactions among estrogen-metabolism genes in sporadic breast cancer. AB - One of the greatest challenges facing human geneticists is the identification and characterization of susceptibility genes for common complex multifactorial human diseases. This challenge is partly due to the limitations of parametric statistical methods for detection of gene effects that are dependent solely or partially on interactions with other genes and with environmental exposures. We introduce multifactor-dimensionality reduction (MDR) as a method for reducing the dimensionality of multilocus information, to improve the identification of polymorphism combinations associated with disease risk. The MDR method is nonparametric (i.e., no hypothesis about the value of a statistical parameter is made), is model-free (i.e., it assumes no particular inheritance model), and is directly applicable to case-control and discordant-sib-pair studies. Using simulated case-control data, we demonstrate that MDR has reasonable power to identify interactions among two or more loci in relatively small samples. When it was applied to a sporadic breast cancer case-control data set, in the absence of any statistically significant independent main effects, MDR identified a statistically significant high-order interaction among four polymorphisms from three different estrogen-metabolism genes. To our knowledge, this is the first report of a four-locus interaction associated with a common complex multifactorial disease. PMID- 11404820 TI - Gene mutations in the succinate dehydrogenase subunit SDHB cause susceptibility to familial pheochromocytoma and to familial paraganglioma. AB - The pheochromocytomas are an important cause of secondary hypertension. Although pheochromocytoma susceptibility may be associated with germline mutations in the tumor-suppressor genes VHL and NF1 and in the proto-oncogene RET, the genetic basis for most cases of nonsyndromic familial pheochromocytoma is unknown. Recently, pheochromocytoma susceptibility has been associated with germline SDHD mutations. Germline SDHD mutations were originally described in hereditary paraganglioma, a dominantly inherited disorder characterized by vascular tumors in the head and the neck, most frequently at the carotid bifurcation. The gene products of two components of succinate dehydrogenase, SDHC and SDHD, anchor the gene products of two other components, SDHA and SDHB, which form the catalytic core, to the inner-mitochondrial membrane. Although mutations in SDHC and in SDHD may cause hereditary paraganglioma, germline SDHA mutations are associated with juvenile encephalopathy, and the phenotypic consequences of SDHB mutations have not been defined. To investigate the genetic causes of pheochromocytoma, we analyzed SDHB and SDHC, in familial and in sporadic cases. Inactivating SDHB mutations were detected in two of the five kindreds with familial pheochromocytoma, two of the three kindreds with pheochromocytoma and paraganglioma susceptibility, and 1 of the 24 cases of sporadic pheochromocytoma. These findings extend the link between mitochondrial dysfunction and tumorigenesis and suggest that germline SDHB mutations are an important cause of pheochromocytoma susceptibility. PMID- 11404821 TI - Magnetic resonance imaging of rheumatoid arthritis: the evolution of clinical applications through clinical trials. AB - Powerful techniques are being developed for evaluating rheumatoid arthritis with magnetic resonance imaging (MRI). Much of this development is being driven by the pharmaceutical and biotechnology industries searching for novel therapies for this disease. Accordingly, the imaging tools that ultimately will be used to direct patients to specific therapies and then to monitor treatment effectiveness and safety are currently being refined and validated in rigorous multicenter and multinational clinical trials aimed at gaining regulatory approval of these new therapies. As these trials approach completion, rheumatologists can anticipate an increased demand for expertise and experience in evaluating disease progression and treatment response with these techniques and the emergence of MRI systems specifically designed for this market. The following discussion reviews this novel pathway for evolving imaging techniques for clinical use through clinical drug trials, lists the most promising MRI markers available today for evaluating joint destruction in rheumatoid arthritis, and speculates on how these techniques will find their way into clinical practice. PMID- 11404822 TI - Microvascular involvement in systemic sclerosis: capillaroscopic findings. AB - OBJECTIVE: To show the most representative capillaroscopic findings in patients with systemic sclerosis (SSc). METHOD: Capillaroscopic evaluation was performed with a videomicroscope at 100 times and 200 times magnification. Representative images of the nailbed microcirculation were obtained in 20 healthy subjects and in 75 patients with SSc. RESULTS: Capillary abnormalities were seen at the nailfold in more than 90% of patients with SSc. Capillary involvement included several morphologic changes, which are easily distinguished from the pattern of healthy controls. Architectural disruption of the nailfold microvascular network, enlarged loops, neoformation of capillaries, loss of capillaries, and avascular areas are the main abnormalities detected in SSc patients. CONCLUSIONS: Nailfold capillary microscopy is one of the most valuable tools for the early diagnosis of SSc and related disorders. Rheumatologists and internists should be able to recognize these abnormalities because of their diagnostic relevance. PMID- 11404823 TI - Pulmonary-renal syndrome in systemic sclerosis. AB - BACKGROUND AND OBJECTIVE: Renal failure, pulmonary hypertension, and interstitial lung disease are major causes of morbidity and mortality in systemic sclerosis (SSc). However, the concomitant occurrence of pulmonary hemorrhage associated with acute renal failure in SSc has been rarely described. The present study is the first analysis of pulmonary-renal syndrome in SSc. PATIENT AND METHODS: We present a 44-year-old woman with SSc who died of a fulminant course of acute renal failure associated with diffuse alveolar hemorrhage. We termed this uncommon and fatal complication of SSc scleroderma-pulmonary-renal syndrome (SPRS). A search of the English-written literature yielded reports of 10 additional similar cases. These patients, together with our present case, form the basis of the present analysis. RESULTS: The average age of the patients with SPRS was 46 years. The majority of the patients (80%) were women, and most had diffuse SSc. SPRS occurred an average of 6.4 years after disease onset and was associated with prior fibrosing alveolitis and/or D-penicillamine treatment. Interestingly, normotensive renal failure seems to characterize the scleroderma patients, because 9 of 11 (82%) had normal blood pressure. SPRS bears a poor prognosis: all of the 11 patients (100%) died within 12 months of admission. However, only 60% of the 5 patients for whom we have treatment data received corticosteroids. CONCLUSIONS: Pulmonary-renal syndrome is a rare but fatal complication of SSc. Because the treatment data are scarce and the prognosis is poor, aggressive treatment with pulse corticosteroids, cyclophosphamide, and possibly plasmapheresis is suggested. PMID- 11404824 TI - The prevalence of musculoskeletal pain and fibromyalgia in patients hospitalized on internal medicine wards. AB - OBJECTIVES: To estimate the prevalence of nonarticular pain complaints (chronic widespread pain, chronic localized pain, transient pain) and fibromyalgia in hospitalized patients and to study utilization patterns of health services associated with pain related problems. METHODS: Five hundred twenty-two patients hospitalized on internal medicine wards were enrolled. Data were collected with a questionnaire covering demographic background, information on pain and other symptoms, utilization of health services, and drug consumption. All subjects were classified into four pain groups: those with no pain, transient pain, chronic regional pain, and chronic widespread pain. Tenderness was assessed by thumb palpation, and patients were diagnosed as having fibromyalgia if they met the 1990 American College of Rheumatology criteria. RESULTS: Sixty-two percent of the patients reported pain; 36% reported chronic regional pain, 21% reported chronic widespread pain, and 5% reported transient pain. Fifteen percent of all patients had fibromyalgia, most of whom (91%) were women. The prevalence of chronic widespread pain and of fibromyalgia in women increased with age. Sleep problems, headache, and fatigue were highly prevalent, especially among those with chronic widespread pain. Patients with chronic widespread pain reported more visits to family physicians (6.2 visits per year) and more frequent use of drugs. They also were more frequently referred to rheumatologists, and they reported more hospitalizations. CONCLUSIONS: Pain syndromes and related symptoms are prevalent among hospitalized patients on the medicine wards. The internist taking care of these patients should be aware of the presence of these syndromes and realize that some of the reported symptoms are partly related to these (undiagnosed) pain syndromes rather than to the cause of hospitalization. PMID- 11404825 TI - Lupus-specific autoantibodies in concomitant human immunodeficiency virus and systemic lupus erythematosus: case report and literature review. AB - OBJECTIVES: To report a case of systemic lupus erythematosus (SLE) in a patient with human immunodeficiency virus (HIV) infection. We also reviewed the medical literature for similar cases to assess the role of "lupus-specific antibodies" in the diagnosis of SLE in the presence of HIV infection. METHODS: We described the presentation, clinical course, and serologic studies of our patient and reviewed the English language medical literature from 1966 to 1999 using MEDLINE with the keywords "SLE," "HIV," "acquired immune deficiency syndrome (AIDS)," and "autoantibodies." RESULTS: Our patient with long-standing HIV infection presented with fever, arthralgias and arthritis, photosensitive rash, oral ulcers, alopecia, headache, pleuritic chest pain, and lymphadenopathy. Laboratory testing showed leukopenia, thrombocytopenia, antibodies against ribonucleoprotein, Smith, and ribosomal-P, as well as immunoglobulin G anticardiolipin antibodies. Review of the literature revealed only 25 cases of concomitant SLE and HIV. In these cases, rheumatologic signs and symptoms were common in HIV and overlapped significantly with SLE. Autoantibodies also occurred frequently in both diseases. CONCLUSIONS: There are very few reported cases of concomitant HIV and SLE. These diseases may be largely mutually exclusive, but distinguishing the two can be difficult because of the high degree of rheumatic complaints and autoantibodies in HIV-positive patients. As illustrated by the patient presented, SLE should be considered in HIV-positive patients with rheumatologic complaints. PMID- 11404826 TI - Use of exogenous estrogens in systemic lupus erythematosus. AB - OBJECTIVE: To review the current literature on the safety of using exogenous estrogens in patients with systemic lupus erythematosus (SLE). METHOD: A MEDLINE search for articles published between 1970 and 2000 regarding the relationship between estrogens and SLE was performed. Emphasis was put on human studies, treatment trials, and epidemiologic surveys. RESULTS: The use of exogenous estrogens in healthy women increases the risk of SLE development. For patients with established SLE, a hypoestrogenemic state appears to be protective against severe flares, whereas exogenous estrogen administration or hyperestrogenemia induced by hormonal manipulation may exacerbate the disease in certain individuals. Both the use of oral contraceptives and the use of hormonal replacement therapy (HRT) increase the chance of venous thromboembolism. The presence of antiphospholipid antibodies may aggravate the risk of thrombosis in SLE. In retrospective studies, HRT appears to be well tolerated in postmenopausal SLE patients. CONCLUSIONS: There are no prospective data that show a deleterious effect of exogenous estrogens on disease activity in human SLE. Oral contraceptives may be considered for patients with SLE in the absence of active nephritis or antiphospholipid antibodies. The slight increase in venous thromboembolic risk should not be the chief deterrent to the use of HRT in postmenopausal SLE patients, considering its various health benefits. PMID- 11404827 TI - Coronary artery disease in systemic lupus erythematosus: A review of the literature. AB - CONTEXT: Coronary artery occlusive disease is a common though underappreciated complication of systemic lupus erythematosus (SLE), typically a disease of young women. A case of a premenopausal patient with SLE and an acute myocardial infarction is presented, and the etiology and management of coronary artery disease in SLE reviewed. OBJECTIVES: To review the incidence, risk factors, pathology and treatment of coronary artery disease in systemic lupus erythematosus. DATA SOURCES: MEDLINE search of articles in English-language journals from 1980 to 2000. The index words "systemic lupus erythematosus" and the following co-indexing terms were used: "coronary artery disease," "atherosclerosis," "vasculitis," "anticardiolipin antibodies," "antiphospholipid syndrome." SELECTION SYNTHESIS AND ABSTRACTION: Papers identified were reviewed and abstracted by the authors with a presentation of a summary. RESULTS: The prevalence of coronary artery disease among women with SLE between the ages of 35 and 44 years is at least 50-fold greater than among age-matched control subjects. Of these, coronary atherosclerosis accounts for the vast majority of cases; vasculitis of the coronary arteries and other causes generally believed to be more typical of SLE are comparatively rare. CONCLUSIONS: The evidence suggests that SLE is a significant risk factor for coronary atherosclerosis independent of the classic risk factors of hypertension, tobacco use, and hyperlipidemia. PMID- 11404828 TI - Possible androgenic/anti-androgenic activity of the insecticide fenitrothion. AB - To date, within the field of endocrine disruption, much focus has been placed on chemicals that mimic oestrogens (so-called xenoestrogens), and the number of such chemicals apparently detected continues to grow steadily. Less effort has been expended on investigating chemicals that mimic, or antagonize, other hormones. Nevertheless, a number of chemicals have been reported to have a weak affinity for the androgen receptor, all of which have, to date, been found to have anti androgenic activity in vivo. In this report, we present evidence that the insecticide fenitrothion can interact with the androgen, but not with the oestrogen, receptor. Using recombinant yeast expressing the human androgen receptor, we found that fenitrothion behaved as an androgen agonist in vitro when tested alone, and that it could antagonize the androgen DHT when both chemicals competed for the androgen receptor in vitro. In vivo studies using both intact and castrated male rats showed no conclusive androgenic or anti-androgenic responses. Changes in organ weights suggestive of anti-androgenic effects were mitigated against by the reduced body weights of fenitrothion-treated rats. The toxicity of the compound precluded the use of higher dose levels to substantiate any tentative findings. Interestingly, fenitrothion (and related insecticides) is structurally similar to flutamide, an anti-androgen used clinically that gives clearly positive responses in both intact and castrated rats. PMID- 11404829 TI - One-year dog toxicity study of D-002, a mixture of aliphatic alcohols. AB - D-002 is a mixture of high-molecular-weight aliphatic alcohols, obtained from bees wax (Apis mellifera), with mild anti-inflammatory properties and effective anti-ulcer activities demonstrated in experimental models. This study investigated the oral toxicity of D-002 administered for 1 year to beagle dogs. Twenty-four beagle dogs (12 males and 12 females) were distributed randomly in three experimental groups (four animals per group): a control and two treated groups received D-002 at 50 and 250 mg kg(-1) (7 days/week) by gastric gavage. Overall, D-002 was well tolerated throughout the study. No signs or symptoms of toxicity were observed, and no mortality occurred during the study. All groups showed similar weight gain and food consumption. No hematological, blood biochemical or histopathological disturbances attributable to treatment were observed. This study shows no drug-related toxicity induced by long-term administration of up to 250 mg kg(-1) D-002 to beagle dogs. PMID- 11404830 TI - Chronic toxicity/oncogenicity study of styrene in CD-1 mice by inhalation exposure for 104 weeks. AB - Groups of 70 male and 70 female Charles River CD-1 mice were exposed whole body to styrene vapor at 0, 20, 40, 80 or 160 ppm 6 h per day 5 days per week for 98 weeks (females) or 104 weeks (males). The mice were observed daily; body weights, food and water consumption were measured periodically, a battery of hematological and clinical pathology examinations were conducted at weeks 13, 26, 52, 78 and 98 (females)/104 (males). Ten mice of each gender per group were pre-selected for necropsy after 52 and 78 weeks of exposure and the survivors of the remaining 50 of each gender per group were necropsied after 98 or 104 weeks. An extensive set of organs from the control and high-exposure mice were examined histopathologically, whereas target organs, gross lesions and all masses were examined in all other groups. Styrene had no effect on survival in males. Two high-dose females died (acute liver toxicity) during the first 2 weeks; the remaining exposed females had a slightly higher survival than control mice. Levels of styrene and styrene oxide (SO) in the blood at the end of a 6 h exposure during week 74 were proportional to exposure concentration, except that at 20 ppm the SO level was below the limit of detection. There were no changes of toxicological significance in hematology, clinical chemistry, urinalysis or organ weights. Mice exposed to 80 or 160 ppm gained slightly less weight than the controls. Styrene-related non-neoplastic histopathological changes were found only in the nasal passages and lungs. In the nasal passages of males and females at all exposure concentrations, the changes included respiratory metaplasia of the olfactory epithelium with changes in the underlying Bowman's gland; the severity increased with styrene concentration and duration of exposure. Loss of olfactory nerve fibers was seen in mice exposed to 40, 80 or 160 ppm. In the lungs, there was decreased eosinophilia of Clara cells in the terminal bronchioles and bronchiolar epithelial hyperplasia extending into alveolar ducts. Increased tumor incidence occurred only in the lung. The incidence of bronchioloalveolar adenomas was significantly increased in males exposed to 40, 80 or 160 ppm and in females exposed to 20, 40 and 160 ppm. The increase was seen only after 24 months. In females exposed to 160 ppm, the incidence of bronchiolo alveolar carcinomas after 24 months was significantly greater than in the controls. No difference in lung tumors between control and styrene-exposed mice was seen in the intensity or degree of immunostaining, the location of tumors relative to bronchioles or histological type (papillary, solid or mixed). It appears that styrene induces an increase in the number of lung tumors seen spontaneously in CD-1 mice. PMID- 11404831 TI - Effects of pond water, sediment and sediment extract samples from New Hampshire, USA on early Xenopus development and metamorphosis: comparison to native species. AB - In an effort to assess potential ecological hazards to amphibian species in selected regions within New Hampshire, the traditional Frog Embryo Teratogenesis Assay-Xenopus (FETAX), a 14-/21 day tail resorption thyroid disruption assay and >30 day limb development tests were conducted with representative surface water and sediment samples. Two separate sets of samples collected from five sites were evaluated. The primary objectives of the study were to determine if samples were capable of inducing early embryo-larval maldevelopment, to determine if maldevelopment included limb defects, to determine if thyroxine co-administration altered the rates of limb malformation and to evaluate the impact of the samples on growth rates, developmental progress and metamorphic climax. Results from these studies suggested that pond water and sediment extract samples, but not whole sediment samples, from B2, FW, LP and W ponds were capable of inducing abnormal early embryo-larval development. In addition, water samples from B2 and W ponds induced significant abnormal hindlimb development. Some abnormal forelimb development was noted in the tail resorption studies, but not to the same extent as the hindlimbs. Each of the water samples induced appreciable developmental delay, including the paired reference site B1, which could be reversed by the addition of exogenous thyroxine. PMID- 11404832 TI - Production of superoxide anion, lipid peroxidation and DNA damage in the hepatic and brain tissues of rats after subchronic exposure to mixtures of TCDD and its congeners. AB - In this study the induction of oxidative stress in the hepatic and brain tissues of rats after subchronic exposure to various mixtures of 2,3,7,8 tetrachlorodibenzo-p-dioxin (TCDD) and two of its congeners, namely 2,3,4,7,8 pentachlorodibenzofuran (PeCDF) and 3,3',4,4',5-pentachlorobiphenyl (PCB 126) was investigated. Four mixtures of TCDD and its congeners, corresponding to 10, 22, 46 and 100 ng of toxic equivalence (TEQ) kg(-1) day(-1), were administered to groups of rats for 13 weeks. The animals were sacrificed at the end of the exposure period and the biomarkers of oxidative stress, including the production of superoxide anion, lipid peroxidation and DNA single-strand breaks (SSBs), were determined in the hepatic and brain tissues. All mixtures caused dose-dependent increases in the production of superoxide anion, lipid peroxidation and DNA SSBs in both tissues, with significantly higher damage in the hepatic compared with the brain tissues. The 22 ng TEQ dose level (TEQ = 22) contains TCDD, PeCDF and PCB 126 at levels that correspond to 7.3, 14.5 and 73.3 ng kg(-1) day(-1), respectively, and it produced effects that correspond to ca. 50% of the maximal production of superoxide anion, lipid peroxidation and DNA SSBs in the hepatic and brain tissues of those animals. Relative to the doses that are required to produce 50% of the maximal production of the biomarkers of oxidative stress by the individual congeners in hepatic and brain tissues of rats, the concentrations of the congeners in TEQ = 22 did result in significant interactivity, probably in the form of additive effects in the hepatic but not in brain tissues. PMID- 11404833 TI - Ozone induces impairment of adrenergic relaxation in thoracic guinea pig tracheas. AB - The effect of ozone on airway relaxation mechanisms has been seldom studied. We exposed guinea pigs to ozone (0.3, 0.6 or 1.2 ppm for 4 h) and studied the in vitro tracheal responses to isoproterenol and nitroprusside, looking for differences between cervical and thoracic regions. We found that ozone did not alter responses to nitroprusside, whereas it produced a concentration-dependent decrease in the responses to isoproterenol in thoracic but not cervical tracheas. Thus, in isolated organ studies care should be taken to avoid mixing cervical and thoracic tracheal regions, at least when adrenergic responses are to be evaluated. PMID- 11404834 TI - Detoxification of lantana hepatotoxin, lantadene A, using Alcaligenes faecalis. AB - Detoxification of lantadene A (LA), the hepatotoxin from Lantana camara var. aculeata, by the bacterial strain Alcaligenes faecalis has been investigated. Lantadene A induced hepatotoxicity concomitant with increases in plasma bilirubin, blood plasma enzymes and histopathological lesions that typify lantana toxicity. The extract of fermentation broth in which LA was incubated with A. faecalis did not elicit any alterations in blood enzyme prolife or liver histopathology, which were comparable with the control group. It is concluded that A. faecalis detoxified LA and no noxious product was formed on incubation of LA with A. faecalis. PMID- 11404835 TI - Effects of caffeine on locomotor activity of horses: determination of the no effect threshold. AB - Caffeine is the legal stimulant consumed most extensively by the human world population and may be found eventually in the urine and/or blood of race horses. The fact that caffeine is in foods led us to determine the highest no-effect dose (HNED) of caffeine on the spontaneous locomotor activity of horses and then to quantify this substance in urine until it disappeared. We built two behavioural stalls equipped with juxtaposed photoelectric sensors that emit infrared beams that divide the stall into nine sectors in a 'tic-tac-toe' fashion. Each time a beam was interrupted by a leg of the horse, a pulse was generated; the pulses were counted at 5-min intervals and stored by a microcomputer. Environmental effects were minimized by installing exhaust fans producing white noise that obscured outside sounds. One-way observation windows prevented the animals from seeing outside. The sensors were turned on 45 min before drug administration (saline control or caffeine). The animals were observed for up to 8 h after i.v. administration of 2.0, 2.5, 3.0 or 5.0 mg caffeine kg(-1). The HNED of caffeine for stimulation of the spontaneous locomotor activity of horses was 2.0 mg kg( 1). The quantification of caffeine in urine and plasma samples was done by gradient HPLC with UV detection. The no-effect threshold should not be greater than 2.0 microg caffeine ml(-1) plasma or 5.0 microg caffeine ml(-1) urine. PMID- 11404836 TI - Determination of the oestrogenic (uterotrophic) activity of extracts of 'general purpose polystyrene (GPPS)' using immature female rats. AB - In Japan there is growing concern about the possible adverse effects of consumption of food from styrene containers (mainly those made from polystyrene paper) due to the alleged oestrogenic activity of styrene oligomers (dimers and trimers), which may migrate into the food. To examine the possible oestrogenic activity of styrene dimers and trimers, extracts were made from 'general purpose polystyrene (GPPS)' and administered orally to immature female rats over a 4 day period. Increase of uterus weight (wet and blotted) was used for assessment of possible oestrogenic activity. To establish the sensitivity of the test method, immature rats were treated with diethylstilboestrol (DES), a well-known oestrogenic compound. It was found that treatment of rats with levels of up to 60 microg of styrene dimers and 930 microg of styrene trimers per kilogram body weight per day did not give any statistically significant increase of the uterus weight (wet or blotted), whereas DES caused statistically significant, dose related increases in uterus weight at levels as low as 0.89 microg kg(-1) body weight day(-1). It was concluded that, compared with the estimated maximum human daily intake of styrene trimers of 1 microg kg(-1) body weight day(-1) from polystyrene food containers, the risk of adverse human health effects with respect to oestrogenicity may be considered negligible. PMID- 11404837 TI - Contribution to the penetration of radionuclides across the skin. Concentration dependence of strontium through the skin in vitro. AB - Toxicological studies of radionuclide passage across the skin, which represents a crucial barrier of radiation, are important for ensuring the quality of the environment. Both (137)Cs and (90)Sr are most frequently involved in radionuclide contamination of the human body. In our study, we selected (90)Sr because this radionuclide is chemically very close to the bio-element calcium. The permeation of (90)Sr from donor solution across the intact skin of 5- or 9-day-old rats (5DR, 9DR) and across stripped and split skin of the 5DR was studied. The experiments in vitro were carried out using vertical diffusion cells. Strontium chloride (SrCl(2)) was used as carrier in the donor solution in different concentrations. Liquid scintillation spectrometry was applied for radiation detection. The experiments showed that: the permeated fraction of (90)Sr(2+) was indirectly proportional to the carrier concentration in the donor solution; the stratum corneum was found to be the principal penetration barrier of strontium; and in the case of the 9DR the dominant route of strontium penetration was along the follicles. PMID- 11404838 TI - Investigation of synthetic peptide hormones by liquid chromatography coupled to pneumatically assisted electrospray ionization mass spectrometry: analysis of a synthesis crude of peptide triptorelin. AB - Triptorelin, a synthetic peptide hormone used in the treatment of prostate cancer by means of reduction in the action of male hormone testosterone, is studied here. The synthetic procedure commonly results in unwanted side products that require extensive purification and characterization of the synthesis mixture. The chromatographic separation of triptorelin from the crude mixture was developed by applying the linear solvation energy relationship (LSER) methodology previously developed, to optimize the composition of the mobile phase in order to avoid lengthy empirical optimization procedures. Electrospray ionization mass spectrometry coupled to liquid chromatography (LC/ES-MS) was used to obtain reliable information on the inevitable side products. The knowledge of the identity of these impurities allows fast optimization of the synthetic procedure and also the therapeutic use of triptorelin peptide hormone. PMID- 11404839 TI - Electrospray ionization detection of inherently nonresponsive epoxides by peptide binding. AB - A small organic molecule that is inherently nonresponsive to electrospray analysis, 1,3-butadiene diepoxide, was analyzed via electrospray ionization (ESI) by binding it to various peptides and observing the product at the characteristic mass shift. The epoxide reacted only with peptides with arginines in their sequence, most likely through a base-catalyzed ring opening to form a covalently bound product. A calibration curve linear over 3 orders of magnitude was generated for the butadiene diepoxide/peptide adduct. Several other epoxides were also reacted with the peptide of choice (angiotensin II), and adducts of these epoxides with the peptide were observed as well, demonstrating the versatility of this method for the analysis of small epoxides. This study demonstrates the possibility of assaying epoxides bound to peptides or proteins in biological samples. Furthermore, it demonstrates an important concept that could be applied to other analytical problems in electrospray: the ability to react an analyte that is nonresponsive to electrospray analysis with an analyte well suited for the technique, and accomplish quantitation based on the adduct formed between the two. PMID- 11404840 TI - Factors determining the performance of triple quadrupole, quadrupole ion trap and sector field mass spectrometers in electrospray ionization tandem mass spectrometry of oligonucleotides. 1. Comparison of performance characteristics. AB - The performance of triple-stage quadrupole (TSQ), quadrupole ion trap (QIT), and double focusing sector field (DFSF) mass spectrometers for the generation of fragment ions to obtain sequence information about oligonucleotides was compared. Upon electrospray ionization (ESI), the charge-state distribution of candidate precursor ions not only varied significantly with the type of mass spectrometer, but also with the size and sequence of the investigated oligonucleotides. While concentration limits of detection for an octanucleotide were in the 100 pmol/L range on the QIT and in the 5-10 nmol/L range on the TSQ and DFSF instruments, those of a 24-mer were in the 2-13 nmol/L range on all three instruments. Reproducibility of mass determination, an important prerequisite for reliable identification of fragment ions, was highest on the TSQ with 0.0037% relative standard deviation over three days. Finally, the tandem mass spectra of a dimethoxytritylated pentanucleotide recorded on the three instruments were compared. Relatively simple spectra dominated by complete series of fragment ions of the (a-B) and w type were obtained on the QIT. Complete series of (a-B) and w ions were also observed on the TSQ. However, additional fragments belonging to the b, c, d, x and z series were found in the spectrum. In the spectrum recorded after in-source fragmentation in the DFSF, only fragments corresponding to the loss of a nucleobase and a complete series of w ions were observed. All three mass spectrometers were suitable for the generation of fragment ions, from which the complete nucleotide sequence of the pentanucleotide could be deduced. PMID- 11404841 TI - Factors determining the performance of triple quadrupole, quadrupole ion trap and sector field mass spectrometer in electrospray ionization mass spectrometry. 2. Suitability for de novo sequencing. AB - The sequence coverage by fragment ions resulting from collision-induced dissociation in a triple stage quadrupole (TSQ) and a quadrupole ion trap (QIT) mass spectrometer of 10-20-mer oligonucleotides was investigated. While (a-B) and w ion series were the most abundant on both instruments, additional ion series of sequence relevance were preferably formed in the TSQ. Thus, a total number of 83 fragment ions were used to deduce the complete sequence of a 10-mer oligonucleotide of mixed sequence from a tandem mass spectrum recorded on the TSQ. The complete sequence was also encoded in the 28 fragments that were obtained from the QIT under comparable fragmentation conditions. Spectrum complexity increased considerably at the cost of signal-to-noise ratio upon fragmentation of a 20-mer oligonucleotide in the TSQ, whereas spectrum interpretation with longer oligonucleotides was significantly more straightforward in spectra recorded on the QIT. The extent of fragmentation had to be optimized by appropriate setting of collision energy and choice of precursor ion charge state in order to obtain full sequence coverage by fragments for de novo sequencing. Moreover, full sequence information was also dependent on base sequence because of the low tendency of backbone cleavage at thymidines. Tandem mass spectrometry on the QIT yielded redundant information that was successfully utilized to deduce the complete sequence of 20-mer oligonucleotides with high confidence. PMID- 11404842 TI - Melanogenesis by tyrosinase action on 3,4-dihydroxyphenylalanine (DOPA) in the presence of polyethylene glycol: a matrix-assisted laser desorption/ionization mass spectrometric investigation. AB - The enzymatic reaction between DOPA and tyrosinase, the enzyme considered to be responsible for melanogenesis, was carried out in the presence of polyethylene glycol (PEG). This choice was made in order to increase the solubility of melanins, since these polymers are highly insoluble. The reaction mixtures were sampled at different times, immediately ultrafiltered to remove the enzyme, lyophilized, and analyzed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The results were very different from those obtained in the absence of PEG. Only a few oligomers of dihydroxyindole (DHI) and dihydroxyindole 2-carboxylic acid (DHICA) were detected in low abundances, whereas new species originating from reaction of PEG with species belonging to the Raper-Mason pattern appeared. The results show that, in the presence of PEG, tyrosinase catalyzed oligomerization of DOPA exhibits kinetics slower than those observed in the absence of the polymer. However, melanogenesis still takes place in the presence of PEG, as indicated by the formation of black pigments and by the detection of DHI and DHICA oligomers, considered to be the first intermediates in melanin formation. PMID- 11404843 TI - Detection of bacteria from biological mixtures using immunomagnetic separation combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. AB - A rapid method for identifying specific bacteria from complex biological mixtures using immunomagnetic separation coupled to matrix-assisted laser desorption/ionization time-of-flight mass spectrometry has been developed. The technique employs commercially available magnetic beads coated with polycolonal antibodies raised against specific bacteria and whole cell analysis by MALDI-MS. A suspension of a bacterial mixture is mixed with the immunomagnetic beads specific for the target microorganism. After a short incubation period (20 mins) the bacteria captured by the beads are washed, resuspended in deionized H(2)O and directly applied onto a MALDI probe. Liquid suspensions containing bacterial mixtures can be screened within 1 h total analysis time. Positive tests result in the production of a fingerprint mass spectrum primarily consisting of protein biomarkers characteristic of the targeted microorganism. Using this procedure, Salmonella choleraesuis was isolated and detected from standard bacterial mixtures and spiked samples of river water, human urine, and chicken blood. PMID- 11404844 TI - Evaluation of several solid phase extraction liquid chromatography/tandem mass spectrometry on-line configurations for high-throughput analysis of acidic and basic drugs in rat plasma. AB - Several configurations using 6- and 10-port switching valves were studied for high flow, on-line extraction of rat plasma coupled to an electrospray triple quadrupole mass spectrometer. Each plasma sample was diluted 1:1 with an aqueous internal standard solution. The sample was injected into a 2.1 x 20 mm cartridge column packed with 25 microm divinylbenzene/N-vinylpyrrolidone packing using 100% aqueous mobile phase at 4 mL/min. After sample loading and sample cleanup, the analytes were eluted from the extraction column with a 1.0-min gradient at 0.4 mL/min. The samples were either analyzed directly after elution from the extraction column or after additional separation using a short high performance liquid chromatography (HPLC) column. The different configurations were tested using an acidic drug (diflunisal) and a basic drug (clemastine) in rat plasma. On line analysis was performed by injecting 200 microL of diluted plasma. The mass spectrometer was operated in the multiple reaction monitoring (MRM) mode. All calibration standards gave relative standard deviations (RSDs) below 5%. The total time per sample was 3 min. PMID- 11404845 TI - Software (MSPECTRA) for automatic interpretation of triacylglycerol molecular mass distribution spectra and collision induced dissociation product ion spectra obtained by ammonia negative ion chemical ionization mass spectrometry. AB - Rapid analysis of molecular mass distributions of triacylglycerol (TAG) mixtures and regioisomeric structures of selected molecular mass species is possible using ammonia negative ion chemical ionization mass spectrometry utilizing sample introduction by direct exposure probe. However, interpretation of spectra and calculation of results is time consuming, thus lengthening the total analysis time. To facilitate result calculation a software package (MSPECTRA 1.3) was developed and applied to automatic processing of triacylglycerol molecular mass distribution spectra and collision induced dissociation (CID) product ion spectra. The program is capable of identifying triacylglycerol molecular mass species possessing different ACN:DB (acyl carbon number:number of double bonds) ratios on the basis of m/z values of [M - H](-) ions. In addition to such identification the program also corrects spectra for abundances of naturally occurring (13)C isotopes and calculates relative proportions of triacylglycerol molecular species in the analyzed samples. If several replicate spectra are processed simultaneously the program automatically calculates an average and standard deviation of relative proportions of molecular species. In the case of CID spectra the program identifies fatty acid fragment ions [RCO(2)](-) and the corresponding [M - H - RCO(2)H - 100](-) ions, and calculates the relative proportions of ions in both groups. These proportions are then used automatically to calculate the fatty acid combinations comprising the parent triacylglycerol molecule and the regiospecific positions of fatty acids. Processing of several replicate product ion spectra simultaneously produces averaged proportions of regioisomers comprising the parent triacylglycerol molecular species and the standard deviation of the analysis. The performance of the program was tested by analyzing triacylglycerol samples of human milk, human milk substitutes, human chylomicron and cocoa butter, and by comparing results obtained by automated processing of the data with manually calculated results. PMID- 11404846 TI - Practical aspects of liquid chromatography/electrospray tandem mass spectrometry for rapid identification of metabolites of a new antifungal agent SYN-2836 in dog urine. AB - This report presents the structural elucidation of 12 urinary metabolites of SYN 2836, a new antifungal agent showing extensive metabolism in beagle dogs, using complementary liquid chromatography/tandem mass spectrometry (LC/MS/MS) methodologies. The 12 SYN-2836 metabolites were readily divided into four groups by considering that all three members of each group, although differing in masses, exhibited highly similar product ion mass spectra. This suggests that the metabolites within each group share a common major substructure. Therefore, all the grouped SYN-2836 metabolites were strategically identified by characterization of the major substructures followed by determination of the additional small substructures. This grouping strategy greatly facilitated the structural elucidation of these metabolites. Other strategies were also employed to achieve as rapid and unambiguous characterization of the SYN-2836 metabolites as possible. PMID- 11404847 TI - Mass spectrometric characterisation of proteins in rennet and in chymosin-based milk-clotting preparations. AB - The protein composition of natural rennet and of chromatographic and crystalline chymosin preparations has been defined by on-line reverse-phase high performance liquid chromatography/electrospray ionisation mass spectrometry (RP-HPLC/ESI-MS) and by tandem mass spectrometry (MS/MS). Natural rennet was found to consist of six chymosin species, corresponding to chymosin A and B genetic variants, each of which comprised a mixture of two other forms differing at theN-terminal end, with one being three residues longer, and the other two residues shorter, than the mature chymosin. Two main tissue proteins were also identified as lysozyme (isozyme 2 plus a novel isozyme labelled 4) and bovine serum albumin. In addition to the proteins, chymosin fragments 247-323 and 288-323 were consistently present in natural rennet. Conversely, chromatographic and crystalline chymosin preparations lacked bovine serum albumin and/or lysozyme, although they contained the same six chymosin species as natural rennet. Since these tissue-specific contaminating proteins each possess specific functions in terms of stabilising enzyme solutions and protecting proteins from proteolytic enzymes, oxidising agents and bacterial proliferation, the rennet may be considered as a functional enzyme preparation that is effectively and naturally adapted to the purposes of cheesemaking. In practice, the highly complex protein composition inherent to natural rennet provided the possibility to differentiate the natural product from other bovine chymosin-based milk-clotting preparations examined in this work. PMID- 11404848 TI - High-speed liquid chromatography/tandem mass spectrometry using a monolithic column for high-throughput bioanalysis. AB - With the ever-increasing workload from a variety of in vitro and in vivo screening procedures, new analytical methodologies to perform bioanalysis in an accurate and high-throughput manner are in great demand. In this work, monolithic columns were used instead of conventional particulate HPLC columns to perform chromatographic separations. Because the pressure drop on a monolithic column was considerably lower than that on a particulate column, a high flow rate (6 mL/min) was used for a 4.6 x 50 mm monolithic column with a total backpressure of about 61 bar measured using acetonitrile/water (50:50). The capability of using a regular column length at high flow rates, combined with the extremely small dependency of separation efficiency on linear flow velocity, allowed for the generation of sufficient chromatographic resolving power in a significantly reduced runtime. As demonstrated in this work, a plasma extract of a mixture of tempazepam, tamoxifen, fenfluramine, and alprozolam were baseline separated within a total analysis time of one minute. An average peak width at half maximum of approximately one second was noted using a generic broad gradient. It was also found that the separation efficiency and signal/noise (S/N) ratios for this separation remained almost constant at flow rates of 1, 3, and 6 mL/min, respectively. The ruggedness of the separation was evaluated by injecting 600 plasma extracts containing the replicates of a standard curve of the above mixture during an overnight run. The chromatographic retention time, separation quality, peak response and sensitivity were highly reproducible throughout the run. This high-speed liquid chromatography/tandem mass spectrometry (LC/MS/MS) system has been used routinely in the authors' laboratory to support drug discovery programs. PMID- 11404849 TI - Structural determination of lysophosphatidylcholines extracted from marine sponges by fast atom bombardment tandem mass spectrometry. AB - A series of lysophosphatidylcholines were isolated from the marine sponge Spirastrella abata by reversed-phase high performance liquid chromatography (HPLC) and analyzed by fast atom bombardment mass spectrometry (FAB-MS). Their structural elucidation was carried out with fast atom bombardment tandem mass spectrometry (FAB-MS/MS). The collision-induced dissociation (CID) of protonated and sodiated molecular ions produced diverse product ions via a series of dissociative processes. Because of the positive charge of the amine group at the end of the molecules, charge-remote fragmentation patterns of specific ions, [M + H](+) or [M + Na](+), were very helpful for the identification of product ions which are characteristic for choline and long hydrocarbon chains substituted at the glycerol back bone. Moreover, the CID-MS/MS spectra of sodium adducted molecular ions for lysophosphatidylcholines yielded common characteristic fragment ions for the choline moiety and characteristic ions [M + Na-103](+), [M + Na-85](+) and [M + Na-59](+) in the higher mass region. PMID- 11404850 TI - FDA is strong advocate for dentistry, patients. PMID- 11404851 TI - Dentistry in 1000 AD. PMID- 11404852 TI - Angioedema: case report. PMID- 11404853 TI - Washington plagued with dental ills. PMID- 11404854 TI - Diagnostic quiz #43. Case no. 1. All of the above. PMID- 11404855 TI - Diagnostic quiz #43. Case no. 2. Buccal bifurcation cysts. PMID- 11404856 TI - Is the indirect crown best? PMID- 11404857 TI - Evolution of enformation in dentistry. PMID- 11404858 TI - Prosthetic procedures for optimal aesthetics in single-tooth implant restorations: a case report. AB - Restoration of a single-tooth implant in the anterior maxilla requires a multidisciplinary approach throughout the treatment and proper communication between the clinician and laboratory technician. Although surgical procedures have an important role in the manipulation of the soft tissues, it is often necessary to perform final contouring of the soft tissue through the provisionalization stage. Unless the soft tissue contour supports the definitive restoration, the treatment is considered a failure. This article reviews various prosthetic procedures that influence the soft tissue contour and establish aesthetics for implant-supported restorations. PMID- 11404859 TI - Cement considerations for implant-supported restorations. PMID- 11404860 TI - Aesthetic restoration of anterior teeth utilizing metal-ceramic crown restorations. PMID- 11404861 TI - Utilization of a small-particle composite resin for anterior and posterior restorations. AB - The ability to acid etch enamel has resulted in the proliferation of adhesive dental techniques that enable conservation of sound tooth structure during the preparation and restorative phases. Recent research has focused on the clinical procedures that use composite resins as well as the material formulations involved in adhesive treatment. This article discusses the physical and optical properties of a composite resin with a smaller filler particle and demonstrates its utilization in anterior and posterior applications as a means of providing functional aesthetic dental care. PMID- 11404862 TI - Stop fooling yourself. PMID- 11404863 TI - Ridge preservation utilizing an alloplast prior to implant placement--clinical and histological case reports. AB - A calcified copolymer alloplast (Bioplant HTR, Bioplant Inc., South Norwalk, CT) was utilized to fill osseous defects in five patients. Hard tissue cores were obtained from the grafted sites and prepared for biopsy. In one patient, the lining of a soft tissue flap was biopsied 8 months postgrafting. Histological evaluation of the specimens revealed that the copolymer particles placed adjacent to the bony defect walls served as an osteoconductive material in which vital remodeling bone and marrow formed and fused to the surface of the particles. This process of bone deposition and remodeling was present 11 years following grafting. PMID- 11404864 TI - Aesthetic success with the utilization of digital imaging. PMID- 11404865 TI - A conscious decision. A review of the use of general anaesthesia and conscious sedation in primary dental care. PMID- 11404866 TI - Letter from the Secretary. Providing dental treatment under GA as a part of a personal dental service (PDS) in the local district general hospital. PMID- 11404867 TI - Chronic obstructive pulmonary disease. AB - COPD is the most common chronic condition in the UK and it varies in severity from mild through to disabling and severe disease with respiratory failure. The treatment of the disease is tailored to the severity of the symptoms and the cornerstones are stopping smoking, inhaled bronchodilator and inhaled corticosteroid therapy. Preoperative assessment of patients with COPD needs to be thorough; remember that these patients may have concomitant ischaemic heart disease. Patients with severe COPD are at particularly high risk when given intravenous sedatives, opiates or general anaesthetics. PMID- 11404868 TI - New perspectives on dentin adhesion--differing methods of bonding. AB - The advent of adhesive dentistry has had a dramatic effect on the practice of aesthetic dentistry, significantly expanding the range of treatment alternatives that clinicians can provide for their patients. Although adhesion is generally associated with the total bonding technique, adhesive systems can be applied in several different manners that include selective, separate, and secure bonding. It is the goal of this article to present the different bonding procedures, to explain the rationales behind each, and to formulate their indications. PMID- 11404869 TI - New directions in modern dentistry. PMID- 11404870 TI - Conservative restoration of compromised posterior teeth with direct composites: a 7-year report. PMID- 11404871 TI - Treatment of traumatic injuries in the front teeth: restorative aspects in crown fractures. AB - Crown fractures are the most common form of traumatic dental injuries encountered in permanent dentition. Restorative treatment modalities incorporate adhesive materials to effectively maintain function and aesthetics. While uncomplicated injuries of the enamel and/or dentin can be treated solely with adhesive procedures, complicated trauma that involves pulp exposure requires the incorporation of a multidisciplinary treatment approach. Fragment reattachment is facilitated by the utilization of bonding agents that enhance retention and aesthetics. This article discusses the application of provisional and permanent restorative options for the treatment of complications following traumatic injuries. PMID- 11404872 TI - Aesthetic considerations for the treatment of partially edentulous patients with removable dentures. AB - Treatment with removable partial dentures (RPDs) is an affordable solution for anterior or posterior tooth loss. In determining a proper treatment solution, it is important for the clinician to consider the patient's aesthetic expectations, socioeconomic situation, and the prognosis for the prosthesis and remaining dentition. This article presents guidelines to optimize the aesthetic result of this treatment and considers framework and clasp concepts; rotational path design; minimal extension of the denture bases; auxiliary attachments or retentive bars; and implants for retention and support of RPDs. PMID- 11404873 TI - Continuing education in the age of the Internet. PMID- 11404874 TI - Treatment of luxation traumatic injuries: definition and classification in the literature. AB - Luxation traumatic injuries affect the hard tissues and may involve periodontal tissues in severe cases. Periodontal healing must be considered during treatment of traumatic injuries that result in total luxation of the teeth. Pulpal healing is essential for the endodontic prognosis of a reimplanted tooth, as well as for its periodontal healing. Luxation injuries can be further classified as intrusive, extrusive, and/or lateral; this categorization further facilitates the mode of splinting and repositioning utilized during treatment. This article reviews various types of dentoalveolar traumatic injuries and discusses commonly incorporated treatment modalities. PMID- 11404875 TI - Teledentistry. An overview. PMID- 11404876 TI - Causes, consequences of dry mouth are on upswing. PMID- 11404877 TI - Tartar control substance may prevent skeletal mineral deposits. PMID- 11404878 TI - Pulpal microvascular response to acute dentinal exposure. PMID- 11404879 TI - Letting presumption outrun gumption. PMID- 11404880 TI - Medical errors in the dental office. PMID- 11404881 TI - The specialist's gift. Response to Ethical Dilemma #32. PMID- 11404882 TI - Oral and maxillofacial pathology case of the month. Calcifying epithelial odontogenic tumor. PMID- 11404883 TI - Restoration of an endodontically treated tooth utilizing a single-unit crown and core system. PMID- 11404884 TI - Strengthening composite resin restorations with ceramic whisker reinforcement. AB - Due to their tendency to fracture, current composite formulations are unsuitable for use in large stress-bearing direct posterior restorations that involve cusps. This study investigated the use of single-crystalline ceramic whiskers for the reinforcement of composite resins. The whisker-reinforced composite materials exhibited physical characteristics (i.e., flexural strength, work-of-fracture, and elastic modulus) that were significantly greater (P < 0.05; Student's t test) than those of traditional composite formulations. The experimental materials also had a surface smoothness that was essentially comparable to hybrid composite control specimens. PMID- 11404885 TI - A century of endodontics. PMID- 11404886 TI - The tipped mandibular molar as a bridge abutment: Part 3--Preparation design. PMID- 11404887 TI - Direct composite resin restoration of adolescent Class IV tooth fracture: a case report. AB - Anterior tooth fractures can be restored utilizing treatment modalities from various dental specialties. The combination of rising patient aesthetic expectations and the desire for conservative dentistry, however, has resulted in the development of intricate direct composite layering techniques. These procedures preserve the natural tooth structure while allowing light and color to be manipulated in the restored dentition. By highlighting the importance of light, color, and material selection, this case presentation documents the use of a direct composite resin technique for the fabrication of aesthetic anterior restorations. PMID- 11404888 TI - The real culprit. PMID- 11404889 TI - Anterior restoration utilizing novel all-ceramic materials. PMID- 11404890 TI - Literary decorum and biased endorsements. PMID- 11404891 TI - Indications and potential of bonded metal-ceramic fixed partial dentures. AB - Recent progress in restorative dentistry has dramatically modified contemporary treatment concepts, which are typified by the preservation of healthy tissues, recognition of aesthetic expectations, and the search for simplified prosthetic solutions. While this shift in focus has justified the increasing interest in implantology, it has also resulted in the expansion of adhesive and conservative treatment options. This article reviews the various means available to replace missing anterior teeth, as well as the periodontal and surgical procedures that are associated with the optimization of biological, functional, and aesthetic results. PMID- 11404892 TI - The evolution of periodontal/implant treatment. PMID- 11404893 TI - The impact of biomaterials and research on dentistry. PMID- 11404894 TI - Increased application of digital radiography for implant therapy. PMID- 11404895 TI - Sequential tooth preparation for aesthetic porcelain full-coverage crown restorations. AB - The fabrication and delivery of functional, biocompatible restorations with a natural appearance have become the definitive goals of restorative dentistry. Although contemporary ceramic materials allow light to be manipulated in restorations to replicate the natural dentition, this objective can be difficult to achieve when tooth preparation is not performed properly. This article presents a sequential approach that can be used to render the proper shape and margin design critical to the fabrication of full-coverage crown restorations that are indistinguishable from the adjacent natural teeth. PMID- 11404896 TI - A century of operative dentistry. PMID- 11404897 TI - The evolution of prosthodontic treatment. PMID- 11404898 TI - Advances in endodontics. AB - Progress in diagnosis, prevention of diseases, treatment of exposed pulp, root canal treatment, retreatment and surgery is reported. Contemporary biological research techniques could set the foundation for a more rational approach to treatment. Pulp inflammation may be prevented by inhibiting bacteria from colonising dentine surfaces. Specific factors could be synthesised to stimulate normal dentine deposition over pulp exposures. Significant improvements have been made in instruments and techniques available for root canal treatment and surgery but there is no clinical evidence that they improve periapical healing. PMID- 11404900 TI - Dentistry 2000--a retrospective. PMID- 11404899 TI - The provision of occlusal splints in primary dental care. AB - Occlusal splints (Michigan splints, night/bite guards or bite-raising appliances) can be an effective, inexpensive and reversible treatment for a wide range of dental problems. OBJECTIVE: The aim of this study was to analyse retrospectively the provision of occlusal splint ('Michigan' type) in general dental practice, following a prescription by a restorative dental consultant. METHOD: One hundred patients were recruited from consultant clinics in a department of conservative dentistry during 1995 and 1996. All patients were prescribed a maxillary, full coverage, heat-cured, acrylic-resin splint (Michigan splint) as part of a treatment plan. An explanatory letter and questionnaire were sent to all patients and to their referring general dental practitioner in 1997 and 1998. RESULTS: A response rate of 79% was achieved in obtaining completed questionnaires from both patients and general dental practitioners. Of respondents 43% (34/79) received an occlusal splint of some form. A small proportion of the respondents (16.5% [13/79]) received a Michigan splint as prescribed. Irrespective of the type of appliance provided, most patients (82% [28/34]) found them helpful. Of those who did not receive an occlusal splint, 38% (17/45) of patients felt financial implications deterred them from obtaining an appliance. Other common reasons for non-provision included: patients felt that symptoms had improved (18% [8/45]) and patients did not agree with treatment (18% [8/45]). The general dental practitioners had similar opinions to their patients. DISCUSSION: These findings raise some serious doubts on the efficacy of consultant clinic advice in the prescription of occlusal splints in general dental practice. The financial and educational issues raised by this study will need to be addressed to improve service provision. CONCLUSION: The results of this study indicate that 16.5% of patient respondents prescribed a Michigan splint at a consultant clinic received such an appliance in general dental practice. PMID- 11404901 TI - Primary dental care in Japan: organisation, management and delivery. AB - The Japanese system of business management and work ethic has been the envy of many in the Western world and many have visited the country to study the methods employed for success. As winner of the Faculty of General Dental Practitioners (UK)'s Stafford-Miller Travelling Scholarship, I spent four weeks in Japan visiting various dental institutions and examining general dental practice. PMID- 11404902 TI - An evaluation of powder-free gloves in general dental practice. AB - PURPOSE OF STUDY: To evaluate the suitability of two non-sterile, powder-free gloves for routine use in general dental practice. BASIC PROCEDURES: Randomly allocated blind evaluation of two powder-free gloves by five dentists and five dental nurses in general dental practice. Each dentist and dental nurse were allocated a four-day supply of two unnamed powder-free gloves. Following glove usage a questionnaire sought their views regarding ease of donning, operator comfort, puncture and tear resistance, and tactile sensitivity. Details of procedure undertaken, time of wear and puncture and tear formation for the dentists' gloves were also recorded. The dentists' gloves were retained and subsequently tested for punctures and tears. MAIN FINDINGS: In terms of ease of donning, operator discomfort, tear resistance and tactile sensitivity neither glove was ideal, however one glove was rated higher than the other. Overall, the dentists noticed only half of the punctures and tears detected in the laboratory. One glove was more prone to punctures with the other glove more liable to tear. Time worn in excess of 20 minutes and endodontic procedures were associated with a higher incidence of punctures. PRINCIPAL CONCLUSIONS: The two powder-free gloves evaluated in this study would appear not to be suitable for routine use in general dental practice. Further developments are required before powder-free gloves of the type evaluated can be expected to receive widespread acceptance for routine use by dentists and dental nurses in general dental practice. PMID- 11404903 TI - The professor, the newspaper and the six-monthly check-up. PMID- 11404904 TI - Primary dental care: time to revise the definition? AB - There have been several definitions of primary dental care over the last 20 years and while each was written to reflect the requirements of the time, most provide a conflicting description. This paper examines past definitions of primary dental care in the context of recent developments in UK dentistry and critically describes possible new definitions. PMID- 11404905 TI - Management of the impacted or displaced maxillary canine. AB - The maxillary canine is commonly displaced or impacted. Monitoring the normal development of the dentition is important to identify and treat potential problems and hence minimise the need for later, more complex therapy. This paper discusses some problems that may occur and the subsequent treatment of these problems, by illustration with five case reports. PMID- 11404906 TI - Three-dimensional shade analysis: perspectives of color--Part II. AB - In addition to outlining the fundamental principles of color and its qualitative and quantitative measurement, the first part of this article explained the chemical, physical, physiological, and psychological realization of color. To complete this discussion, the second part focuses on numerous variables that affect shade determination in the clinical chromatic environment. A contemporary shade selection system is also presented, and its systematic approach and limitations when utilized in a laboratory or clinical environment are detailed. PMID- 11404907 TI - Diagnosis and treatment of burning mouth syndrome. PMID- 11404908 TI - A multidisciplinary approach to complex aesthetic restoration with diagnostic planning. PMID- 11404909 TI - Fourth-generation intraradicular posts for the aesthetic restoration of anterior teeth. AB - Endodontically treated teeth have been traditionally restored with cost-metal posts prior to receiving aesthetic care. Due to the difficulties associated with early posts, resources have been invested to develop biocompatible endodontic posts capable of satisfying functional and aesthetic criteria. These initiatives have resulted in the introduction of carbon-based and glass-fiber posts with translucencies and moduli of elasticity closely approximating that of the dentin for use in adhesive dental procedures. This article demonstrates the rationale and application of translucent glass-fiber posts for the restoration of pulpless teeth. PMID- 11404910 TI - Fixed or removable implant-supported restorations in the edentulous maxilla: literature review. AB - Among the prosthesis designs used to treat the edentulous maxilla are fixed or removable implant-supported restorations. Since the aesthetic requirements and preoperative situation of each patient varies, controversy exists regarding prostheses' success rates and complications. The purpose of this article is to compare the treatment options and prosthesis designs with their indications and to compare implant and prosthesis success and treatment expense. This objective was accomplished through the review of reports with regard to varying design considerations and factors that influence the decision-making process and treatment outcomes. PMID- 11404911 TI - Efficacy of home bleaching systems with and without tray reservoirs. PMID- 11404912 TI - The efficacy of non-opioid analgesics for postoperative dental pain: a meta analysis. PMID- 11404913 TI - Halothane, dental anaesthesia and children. PMID- 11404914 TI - Anaesthetic deaths in dentistry. PMID- 11404915 TI - Placement of direct composite veneers utilizing a silicone buildup guide and intraoral mock-up. AB - The indications for direct composite resins have recently been expanded to include predictable and convenient application in the aesthetic zone. The availability of composite materials with improved physical and optical characteristics facilitates the development of enhanced aesthetics while maintaining vital function. This article presents a simplified technique that combines function with aesthetics by utilizing an intraoral composite mock-up for initial communication and a lingual/incisal silicone stent of the mock-up to transfer the information to the definitive restorative buildup. PMID- 11404916 TI - The blind leading the blind. PMID- 11404917 TI - Posterior restoration using laboratory-fabricated composite resin inlays/onlays. PMID- 11404918 TI - The versatility and enhanced aesthetics of laboratory-fabricated composite resin restorations. PMID- 11404919 TI - Provisionalization for porcelain veneers using bis-acrylate and polyvinylsiloxane matrix. PMID- 11404920 TI - Treatment of teeth with open apices using mineral trioxide aggregate. AB - Injuries to the dentition may ultimately result in the interruption of root development. If the pulp remains vital following trauma, this state should be maintained and root-end closure should be induced by apexogenesis. When the pulp is necrotic, the placement of an apical barrier can be utilized as an alternative to establish an environment that facilitates the closure of the apical opening. This article demonstrates the use of mineral trioxide aggregate as an apical barrier material for root-end closure in the permanent teeth of three patients. PMID- 11404921 TI - Diagnosis and treatment of recurrent aphthous ulceration. PMID- 11404922 TI - Diagnostic quiz #39. Case no. 1. Cherubism. PMID- 11404923 TI - Diagnostic quiz #39. Case no. 2. Granular cell tumor. PMID- 11404924 TI - Basic life support. A reminder. PMID- 11404925 TI - First response. A new course from SAAD. PMID- 11404926 TI - Student elective report. Paediatric sedation in the USA. PMID- 11404927 TI - What is required to practice dental anaesthesia in the UK? PMID- 11404928 TI - Letter from the Secretary. Members of SAAD already know there is a very real need for conscious sedation in the everyday practice of dentistry. PMID- 11404929 TI - Halothane and general anaesthesia in dental treatment. PMID- 11404930 TI - Brietal, an old friend. PMID- 11404931 TI - Diagnostic quiz #47. Case no. 1. Lichen planus. PMID- 11404932 TI - Diagnostic quiz #47. Case no. 2. Idiopathic external or internal resorption. PMID- 11404933 TI - Your vote determines how dentistry is practiced. PMID- 11404934 TI - The truth about expert witnesses. PMID- 11404935 TI - The most overlooked emergency--syncope. PMID- 11404936 TI - Diagnostic quiz #45. Case no. 1. Fibroma, peripheral ossifying fibroma, peripheral giant cell granuloma, pyogenic granuloma. PMID- 11404937 TI - Diagnostic quiz #45. Case no. 2. Central odontogenic fibroma. PMID- 11404938 TI - The principles of money-back guarantees. PMID- 11404939 TI - Diagnostic quiz #46. Case no. 1. Squamous cell carcinoma. PMID- 11404940 TI - Diagnostic quiz #46. Case no. 2. Apical cyst. PMID- 11404941 TI - Intraoral camera helps predict and prevent tooth loss. AB - The video exam has proven to be a most effective means of re-establishing trust between patient and dentist by making the patient a co-diagnostician. In fact, it is not uncommon for patients to stop the diagnosis and ask specific questions about what they see. They can begin to diagnose pathology even before the dentist mentions it. A major benefit to the dentist is that invariably the exam reveals conditions not seen with typical clinical exams even using magnification loops. Although the intraoral camera has been around for more than a decade, its benefit during the video exam is more important now than ever before (8, 9). Every new patient and all patients of record should have the benefit of a video exam, as well as the dentist. PMID- 11404942 TI - Mood disorders in dental patients. PMID- 11404943 TI - Neurophysiology of acute dental pain. PMID- 11404944 TI - A new medication for treatment of dry mouth in Sjogren's syndrome. AB - Traditionally, treatment of dry mouth in SS is focused on palliative measures (using salivary substitutes). However, due to the dynamic nature of the oral cavity, the salivary substitute is removed from the mouth during swallowing. Therefore, the duration of effect of salivary substitutes is short. Another drawback is that salivary substitutes do not provide the protective roles of saliva. Effective treatment of dry mouth requires increasing salivary output. Gustatory stimulation of the salivary glands with sugar free gum and sugar free candies may be effective in inducing salivary output, however, they impose significant inconvenience on the patient which can compromise compliance. Pharmacological stimulants provide an alternative effective measure and improve compliance. Both Salagen and Evoxac are FDA approved salivary stimulants. They are effective, and safe given awareness of their indications, contraindications, potential adverse effects, and patient's tolerance. PMID- 11404945 TI - Dental information a new career in dentistry: a new career in dentistry. PMID- 11404946 TI - The patient's errant checkbook. Response to ethical dilemma #35. PMID- 11404947 TI - The art of cosmetic dentistry. PMID- 11404948 TI - Geriatric problem solving with adhesive dentistry, using the Contour Strip and direct composite placement. AB - The ability to create periodontally sound gingival and non-staining or plaque retaining interproximal margins on spacious carious lesions is a daunting challenge in many situations. The case as presented will illustrate the use of a unique matrix system, The Contour Strip from Vivadent, which will encompass the entire outline margins of the upper right lateral incisor of an elderly patient. The age of the patient has no bearing on the use of the technique, but merely reenforces the value of the system in all other situations mimicking a similar restorative obstacle. The use of this custom shaped matrix band, eliminate the free-hand placement and then the laborious creation and finishing of the restoration. LEARNING OBJECTIVES: After reading this paper the reader will understand the manipulation and use of a new labial matrix band which can totally surround the gingival and proximal margins of any anterior and pre-molar teeth. TECHNIQUE CONSIDERATIONS: There are four rules to follow when using adhesive dentistry: Proper preparation of tooth surfaces, both dentin and enamel through mechanical and/or chemical means. Creation of matrix formed molds. Injecting all resins, from the lowest to the highest viscosity. Polymerizing thoroughly through trans-enamel polymerization thus controlling shrinkage. PMID- 11404949 TI - Oral and maxillofacial pathology case of the month. Osteosarcoma. PMID- 11404950 TI - Tongue piercing: a fad with serious dental and physical consequences. PMID- 11404951 TI - Florida Board of Dentistry shows its teeth to practice-management companies. PMID- 11404952 TI - Diagnostic quiz #44. Case no. 1. Leukemic infiltrate.. PMID- 11404953 TI - Diagnostic quiz #44. Case no. 2. Odontoma. PMID- 11404954 TI - A new mission and strategic plan. CU School of Dentistry. PMID- 11404955 TI - Expanded patient care. PMID- 11404956 TI - Attachment mechanisms for removable implant restorations. PMID- 11404957 TI - Hard and soft tissue management for the restoration of traumatized anterior teeth. AB - The restoration of coronally fractured anterior teeth without surgical invasion is contingent upon several factors. Specifically, the biologic width of the tooth should not be violated by the apical extent of the fracture, and the residual root structure must possess an adequate ferrule. In patients with these conditions, it is possible to prosthetically restore the tooth following orthodontic extrusion. This article describes a technique in which orthodontic extrusion is utilized to provide adequate tooth structure for the prosthetic restoration of patients who presented with fractured anterior teeth. PMID- 11404958 TI - Management of periodontally involved maxillary anterior dentition: a 10-year report. PMID- 11404959 TI - Utilization of the surgical microscope for advanced plastic periodontal surgery. AB - The increased demand for mucogingival aesthetics has required the optimization of periodontal procedures. Microsurgery is a minimally invasive technique that is performed with the surgical microscope and adapted instruments and suture materials. While this hardware and knowledge of various operations are necessary to achieve patient aesthetic expectations, clinicians must be willing to undergo an extended period of systematic training to become familiar with novel operating procedures and instruments. This article describes the application of the surgical microscope to provide enhanced perioplastic treatment. PMID- 11404960 TI - Enhancing physiologic pigmentation utilizing a free gingival graft. PMID- 11404961 TI - Experimenting on patients. PMID- 11404962 TI - Orthodontic extrusion and orthodontic extraction in preprosthetic treatment using implant therapy. AB - It has been well documented that orthodontic extrusion is a valuable adjunct to site preparation prior to implant placement. This treatment modality may also be utilized to salvage and restore an extensively damaged tooth for use as a natural abutment. By controlling the orthodontic phase, the hard and soft tissues can be manipulated into a position that permits aesthetic restoration. This article demonstrates the use of this procedure as an adjunct to site preparation and in conjunction with conventional restorative therapy. PMID- 11404963 TI - Dental erosion: diagnostic-based noninvasive treatment. AB - Lesions that result from dental erosion can be difficult to recognize- particularly when abrasion and attrition are also present. Consequently, dental erosion is often misdiagnosed and mistreated by radical restorative modalities that compromise the vitality of the pulp. This article provides clinicians with knowledge concerning the diagnosis of the complex lesions of dental wear and demonstrates the conservative treatment of this condition. Two cases that exhibit marked tooth wear in anterior teeth and their subsequent restoration utilizing occlusal principles and composite resin are presented. PMID- 11404964 TI - Intraoral cameras--the advanced technology tool of the millennium. PMID- 11404965 TI - Patients, guidelines and the NeLH. PMID- 11404967 TI - 'Too few dentists? Workforce planning 1996-2036'. PMID- 11404966 TI - Going private? Exploring the drift away from the NHS. AB - The perception that dentistry is being privatised is widespread. A two-phase study was undertaken to investigate the factors influencing dentists' decisions to practise in the public and or private sectors. In phase I a national postal survey of general dental practitioners (n = 1011) explored differences in working practices and experience of NHS and non-NHS work. Survey data were supplemented in phase II by in-depth interviews with four subsamples of dentists in two regions. The survey found that although the majority of dentists continued to treat the majority of their patients within the NHS since the introduction of the 1990 contract patterns of practice have changed. Where private dentistry was increasing, it was regionally variable and arose mainly from concerns with financial security, maintaining quality of work and autonomy. Ideological differences were apparent in relation to differences in practice mix. Those who continued to work in the NHS, because of insufficient demand for private dentistry, did so in order to provide access to treatment and to maintain a reliable source of income and pension rights. Dentists interviewed expressed reluctance in withdrawing from NHS practice. Disillusionment with government policy and recognition of the marginal nature of dental health issues suggests that current trends will continue. PMID- 11404968 TI - Advances in oral medicine. AB - This paper reviews some of the more important areas of oral medicine where significant advances have been made in the past decade. There have been developments in the definition of new entities causing orofacial pain, and new therapies. Oral cancer has been recognised as increasing in incidence and the genetic basis is becoming clearer, along with improvements in early diagnostic techniques and treatment. The basis of aphthae is being unravelled and new therapies are appearing. There is greater understanding of the aetiopathogenesis and treatment of various dermatoses that present in the mouth. Treatment and prevention of leukoplakias is improving and the importance of candidosis has become evident. PMID- 11404969 TI - Rates of GDP referral of complete dentures to a teaching hospital. AB - PURPOSE OF STUDY: To identify areas and sites of excessive referral of complete dentures by GDPs, allowing for factors which might distort referral rates. METHODS: Two hundred and six consecutive valid referrals of patients over the age of 50 years with complete dentures to a local dental teaching hospital and originating from the 29 Sheffield electoral wards were audited from the beginning of 1995 to the end of 1997. The referring practitioner and electoral ward were recorded. RESULTS: Referral rates were calculated according to the size of the 50+ population within each ward and were found to be normally distributed, following exclusion of two wards with much higher referral rates than normal (3.5 and 6 SDs [standard deviations] from the mean). After calculating mean numbers referred per full-time equivalent (FTE) GDP for all wards, practices and practitioners originating excessive numbers of referrals within a high-referring ward could be identified. CONCLUSION: After taking into account factors which might permit a high referral rate to be acceptable, practices with inappropriately high referral rates can be identified and remedial teaching need pinpointed. PMID- 11404970 TI - How orthodontic functional appliances work. AB - Orthodontic functional appliances are an essential part of an orthodontist's armamentarium. The amount of their use varies greatly, some feeling they have limited use with minor effects, others claiming dramatic changes in facial growth and appearance, without significant genetic constraints. The evidence on clinical use is not equivocal, the general results however not showing an unlimited amount of skeletal change, but rather a balance of change in the growth of the facial bones and dentition with clinically significant changes in the facial soft tissues. This article examines the current evidence. PMID- 11404972 TI - Bite-protective appliances and the "Save that smile" campaign. PMID- 11404971 TI - Techniques used by a group of United Kingdom-based dental practitioners during the provision of indirect tooth-coloured restorations. AB - A number of techniques involving the use of various instruments, brushes and devices have been described for the removal of excess luting material and subsequent finishing around the margins of indirect tooth-coloured restorations. There is little information available on the clinical techniques employed by UK dental practitioners in the placement of these restorations, or on the effectiveness of these techniques. A questionnaire was designed to elicit such information. This was distributed to 500 UK-based dental practitioners. A total of 301 practitioners returned completed questionnaires, giving a response rate of 60%. The results indicated that 88% of the respondents routinely provided porcelain veneers for their patients, with 94% of these respondents indicating that they routinely prepared teeth during the provision of such restorations. A light-cured resin composite luting material was used by 29% of the respondents in placement of veneers, while 69% used a dual cured composite. Excess uncured luting material was removed by a number of methods and a variety of methods were used by respondents to finish the margins of the veneers after placement. Of the respondents 68% indicated that they routinely provided tooth-coloured inlays and onlays, with 89% indicating that they routinely used a dentine bonding system when luting their indirect tooth-coloured restorations. It is concluded that a high proportion of the practitioners surveyed were providing porcelain veneers and tooth-coloured inlays and that a wide variety of materials and finishing techniques were being used. PMID- 11404973 TI - Diagnostic quiz #40. Case no. 1. Malignant lymphoma. PMID- 11404974 TI - Hepatitis C screenings available at FNDC. PMID- 11404975 TI - Diagnostic quiz #41. Case no. 1. Dentigerous cyst. PMID- 11404976 TI - Diagnostic quiz #41. Case no. 2. PMID- 11404977 TI - Management service organizations: business and legal considerations. PMID- 11404978 TI - Diagnostic quiz #37. Case no. 1. Squamous cell carcinoma. PMID- 11404979 TI - Diagnostic quiz #37. Case no. 2. Malignant lymphoma. PMID- 11404980 TI - Use Web site for marketing. PMID- 11404981 TI - The structure of the dental workforce in the 21st century. AB - Graham Try (page 9) has suggested that the UK is heading towards dentist shortage. This paper picks up the story from there, speculating on how a shortage of dentists might stimulate changes in working methods. The market place will look for workforce configurations which make business sense at practice level and which deliver services in a form which patients find helpful and convenient. Although some dental procedures can be carried out by people with less training than dentists, this may not contribute to economic efficiency and could make the customer service experience less appealing. Changes in the structure of the workforce may therefore be less radical than has sometimes been suggested. Much depends on how decision-making and regulatory processes evolve. It seems likely that local and practice level decision-making will grow at the expense of central planning. PMID- 11404982 TI - Is it better to leave or restore carious deciduous molar teeth? A preliminary study. PMID- 11404983 TI - The impact of advances in dental sciences on primary dental care. AB - The next century will see phenomenal changes especially and, most importantly, in patient empowerment, in the improvement of health in the disadvantaged, and in the harmonisation of standards of healthcare throughout the world. Advances in the dental sciences will facilitate these improvements but the major changes surely will be largely stimulated and motivated by the information revolution and the advances in molecular and materials sciences. The identification, and protection against disease, of patients who, because of a genetic predisposition, are at particular risk of disease is likely to become possible. Simple, sensitive, effective, non-invasive and safe diagnostic procedures will become available, as will chairside diagnostic test kits that would identify specific microorganisms causal of oral disease. The advances in digital imaging will be taken further and linked to record-keeping. Specific safe agents targeted at responsible microorganisms will be devised, and systems to deliver therapeutic agents effectively and prevent tissue damage simply will be perfected. Painless, minimally-invasive techniques for the removal of diseased oral tissues will become available, as will biocompatible materials for replacement of lost oral tissues. PMID- 11404984 TI - A materials scientist's glimpse into the new millennium. AB - Questions about the materials which are in current use are posed and answered in this review, which speculates about the sort of dental materials we will be using, or would like to use in the next century. Consideration is given to the merits and failings of dental amalgam, composites and glass-ionomer cements prior to laying down the requirements of the ultimate direct filling material. The case for the continued use of dental alloys for large restorations is discussed and new ways of forming these are described. The use of many cunning techniques to improve the resistance of dental ceramics to catastrophic failure is reported and a general improvement in both the denture bases and orthodontic appliances of the future is predicted. PMID- 11404986 TI - Rectangular collimation. PMID- 11404985 TI - The impact of the electronic media on primary dental care in the 21st century. AB - This paper attempts to predict how electronic media will impact on primary dental care in the twenty-first century. The predictions for the next 10 years are based on developments currently in train or on the premise that equipment and techniques, which have been recently introduced and are at present used by relatively few, will be used by many by 2010. During this decade, the impact of electronic media is considered in the areas of patient support, diagnosis, treatment planning, provision of treatment and its maintenance, quality control and practice management. The paper then speculates briefly about how dentists in primary dental care might practise in the year 2100. It concludes that although electronic media are likely to play an increasingly important role, a clinician's skills, knowledge and ability to communicate with patients will remain the keys to successful practice for the foreseeable future. PMID- 11404987 TI - Sentinel practices in dentistry: a preliminary evaluation. AB - The objectives of this study were to compare the socioeconomic make-up and the dental caries and dental treatment patterns of a general dental practice (GDP) population of five-year-old children with those of the total resident population of five-year-old children in a specific locality, and, secondly, to examine the process of gathering information on oral healthcare needs in primary dental care. The study was set in Halton, North Cheshire. Data were collected retrospectively from the patient records of four GDP sentinel' practices using a common data abstraction form. The socioeconomic profiles of the GDP population and the 1995/6 NHS child dental health survey population were compared using the Super Profiles geodemographic classification by plotting frequency distributions. The dmft of each population was compared by calculating 95% confidence intervals. The GDP population showed a slight over-representation in the more affluent groupings of the Super Profiles Lifestyle categories and a more dramatic under-representation in two of the more deprived groupings. The confidence intervals for dt and dmft of the GDP data did not include the mean figures produced by the NHS survey, indicating a significant difference at the P < 0.05 level. These differences may be accounted for by the differences in the socioeconomic make-up of the two populations. Local practice policies on patient selection may also have a consequential effect on population disease estimates derived from primary dental care. Data on population disease experience from primary dental care could only be produced by hand-sorting through patient records, which was time-consuming and inefficient. Standardised electronic systems will need to be developed to make GDP data-collection a viable proposition. Information from primary dental care has the potential to make a major contribution to locality oral health needs assessment and it should be seen as being complementary to information gathered from epidemiological surveys. PMID- 11404988 TI - Dental education in the 21st century. AB - The continuum of dental education has been greatly strengthened by the General Dental Council's educational initiatives during the past five years and these are described in this paper: lifelong learning and recertification will reassure the public and full integration of team members through statutory registration with one self-regulatory body is an innovation in the regulation of health professionals. Future training and educational strategies, including the health science concept, for team members are explored. PMID- 11404989 TI - Too few dentists? Workforce planning 1996-2036. AB - The development and use of forecasting models can stimulate the debate on workforce planning for dentists by improving the quality of the data available, by providing a framework within which the debate can be conducted and by providing a means whereby the effects of different trends and policy options can be measured in a consistent way. The number of dentists on the Dentists Register aged under 60 years may well fall over the 40 years between 1996-2036, while the proportion of female dentists is likely to grow from under 30% to almost 50%. If the present gender differences in dentists' output persist, the effective 'whole time equivalent' (WTE) size of the dentist workforce could fall by 15%. While future patterns of disease and the efficacy of future treatments are difficult to predict, the demand for dentistry is very likely to increase, given the forecast ageing of the UK population. The number of people aged 65-74 years could increase by 50% to eight million and those over 75 years by 70% towards seven million. Further, almost of all of this greatly increased older population will have retained some of their own teeth. The edentulous adult population was 37% in 1968 and might fall to less than 5% by 2036. These trends taken together could increase the number of courses of treatment per WTE dentist by as much as 40% over these 40 years. Such modelling can help to reduce the degree of uncertainty in future workforce planning. PMID- 11404990 TI - Diagnostic quiz #38. Case no. 1. Central giant cell granuloma. PMID- 11404991 TI - Clinical considerations in cement selection for provisional restorations--Part I. PMID- 11404992 TI - Achieving aesthetic excellence through an outcome-based restorative treatment rationale. AB - The conceptual basis of restorative-driven implant dentistry is well established among clinicians. Its implementation includes the development of a prosthetic blue-print that will serve as a guide throughout therapy. While these concepts may be applied to prosthetic dentistry in general, their benefits are most compelling in the treatment of the aesthetic zone. This article demonstrates a systematic multidimensional approach for the establishment and incorporation of definitive aesthetic objectives throughout the diagnostic, adjunctive, and restorative treatment phases. PMID- 11404993 TI - Extensive aesthetic rehabilitation with CAD/CAM all-ceramic crown restorations. PMID- 11404994 TI - An innovative chairside bleaching protocol for treating stained dentition: initial results. AB - For years, investigators have attempted to develop a predictable means of bleaching pathologically and nonpathologically stained dentition. While previous efforts have modified the concentration of the bleaching material, the duration of the procedure, and the manner by which the bleaching agent is activated, the ability to affect a significant shade improvement remains an elusive objective. This article demonstrates an innovative technique used to influence the penetration of oxygen ions into the tooth enamel, which may resolve this clinical dilemma. PMID- 11404995 TI - Where are the limits? Esthetic dentistry comes of age! PMID- 11404997 TI - Digital images--pictures & movies. PMID- 11404996 TI - Clinical presentations of stress distribution in teeth and the significance in operative dentistry. AB - Stress distribution in human tooth structure can be visualized through the use of Moire fringes, which has improved the clinical understanding of recently identified anatomical structures in molar occlusal surfaces. This article discusses the concept of a "peripheral rim of enamel" and describes the manifestation of compressive and tensile fractures within the peripheral rim of enamel and dentin. It also emphasizes the benefits of microdentistry techniques and minimally invasive preparation designs in the long-term preservation of the natural tooth structure. PMID- 11404998 TI - Starting the legacy in 2000. PMID- 11404999 TI - Diagnostic quiz #42. Case no. 1. Epithelial dysplasia with hyperkeratosis and focal lichenoid mucositis. PMID- 11405000 TI - Diagnostic quiz #42. Case no. 2. Central mucoepidermoid carcinoma. PMID- 11405001 TI - Connective tissue grafting for root coverage in multiple Class III gingival recessions with enamel matrix derivative: a case report. AB - This case presentation demonstrates an innovative approach to root coverage that uses an enamel matrix derivative in conjunction with periosteal connective tissue grafting in a patient with multiple gingival facial recessions. A 22-year-old female patient presented for the resolution of aesthetic concerns associated with multiple gingival recessions for teeth #23(32) through #25(41). This technique achieved soft tissue coverage of the root surfaces and effectively improved the patient's aesthetic appearance. These satisfactory clinical results have been maintained for 12 months. PMID- 11405002 TI - Connective tissue grafts for the treatment of discolored roots and amalgam tattoos. PMID- 11405003 TI - Flapless anterior implant surgery: a surgical and prosthodontic rationale. AB - Full-thickness periosteal flap surgery is often accompanied by potential marginal bone loss and/or soft tissue recession. This is critical, particularly for the single-unit implant-supported restoration in the anterior maxilla, where the harmony of the soft and hard tissue architecture is of paramount importance to the development of natural aesthetics and function. This article demonstrates a flapless implant surgery technique in the anterior maxilla for optimal aesthetic results. The indications and limitations of this procedure are also discussed. PMID- 11405004 TI - Using a fixed provisional as an orthodontic anchor in forced eruption. PMID- 11405005 TI - Education, research, and faculty in today's dental schools. PMID- 11405006 TI - Beyond nature. PMID- 11405007 TI - Tooth color selection and characterization accomplished with optical mapping. AB - Due to rising patient expectations and recent developments in material composition and clinical techniques, the demand for restorations that replicate the natural dentition has increased significantly. It is imperative that clinicians and dental technicians develop the communication skills necessary to ensure predictable and reliable shade reproduction. This article defines tooth color and its optical properties while describing techniques for the accurate translation of these characteristics into a laboratory prescription form that utilizes optical mapping procedures. PMID- 11405008 TI - Vertical shoulder preparation design for porcelain laminate veneer restorations. AB - The clinical success of porcelain laminate veneers depends on numerous factors that include the framework of the dental and gingival structures and their ability to withstand masticatory forces. In order to ensure aesthetic harmony, the clinician must reproduce numerous anatomical components (e.g., cementoenamel junction, points of concavity, developmental lobes, line angles, incisal edge survey form) in restorations while simultaneously providing sufficient functional integrity. This article demonstrates the vertical shoulder preparation design, which enhances the aesthetics and stabilization factors of porcelain laminate veneer restorations. PMID- 11405009 TI - Cemental tears related to severe localized periodontal disease. PMID- 11405010 TI - Reducing inequalities in oral health. PMID- 11405011 TI - Dental science--reality of the evidence. PMID- 11405012 TI - The reduction of broken appointments in general dental practice: an audit and intervention approach. AB - PURPOSE OF STUDY: Accumulation of time lost because of broken appointments in general dental practice can lead to an inefficiently-run practice and may also lead to reduced financial income. This study assessed the reasons for patients failing to attend and analysed the effect of two simple interventions in reducing the numbers of broken appointments. BASIC PROCEDURES: This study audited the causes of broken appointments in a Glasgow dental practice over a four-month period and, in a second four-month period, analysed the effect of using two new methods of patient information in reducing the number of broken appointments in two target groups of the practice population. MAIN FINDINGS: The majority of patients failing to attend for dental treatment were unemployed. Simple changes to recall letters and appointment cards significantly reduced the rates of failed appointments in patients attending the practice. PRINCIPAL CONCLUSIONS: Simple modification to existing practice systems (recall letters and appointment cards) led to a statistically significant reduction in the number of broken appointments. PMID- 11405013 TI - Advances in periodontology. AB - Advances in periodontal science and practice over the last decade have radically changed the understanding of periodontal diseases and have opened new, exciting prospects for both medical and surgical therapy of periodontal diseases. Establishment of the aetiology and pathogenesis of periodontitis, understanding of the unique genetic and environmental susceptibility profile of affected subjects, and recognition of the systemic implications of periodontal infections are the key research findings. The use of randomised, controlled, clinical trials has allowed the development of evidence-based periodontology. Adjunctive antimicrobial therapy, regenerative periodontal surgery, periodontal plastic surgery, bone regeneration surgery in the light of implant treatment, and advanced soft tissue management at implant sites have radically changed practice. PMID- 11405014 TI - The professor, the newspaper and the six-monthly check-up. PMID- 11405015 TI - An exchange of e-mails. PMID- 11405016 TI - An analysis of the qualitative factors which influence young people's acceptance of orthodontic care. AB - INTRODUCTION AND METHOD: The qualitative data volunteered as responses to open questions in a self-administered questionnaire study were analysed to identify the psychological factors influencing young people's behaviour in relation to orthodontic care and to gain an understanding of the psychological factors which influence adolescents' acceptance of orthodontic care. The study was conducted in all Walsall and Dudley secondary schools. RESULTS AND DISCUSSION: The responses of the young people who were in year 10 of education with an average age of 15.0 years demonstrated the importance of personal constructs, peer group and media influences, parental influences, conflicting messages, teasing, symptom perception, appearance and self-image, and interpersonal relationships in determining whether or not young people either seek and accept or reject orthodontic treatment. CONCLUSION: The study concluded that it is essential that clinicians involve patients fully and honestly in discussions concerning their orthodontic therapy in order to enable them to make a considered consent. PMID- 11405017 TI - Characteristics of children attending for dental general anaesthesia in 1993 and 1997. AB - OBJECTIVES: To confirm clinical observations and to investigate the characteristics (demography, dental anxiety status, dental health status, treatment experiences and dental health behaviours) of children attending for dental general anaesthesia (DGA). DESIGN: A case series design was used in which each consecutive parent and child, referred for DGA, were invited to take part. METHOD: Identical protocols for the selection of the sample, administration of the questionnaire and clinical examination were used to ensure comparability. To account for seasonal variation the data were collected during and between the months of January and May in both 1993 and 1997. Two hundred children and their parents were selected in 1993 and 200 in 1997. The parent completed a questionnaire. It enquired of the parent and child demography, parental dental anxiety status, and the child's treatment experiences. Children were asked to complete the Children's Fear Survey Schedule (CFSS) to assess their dental anxiety. The child's caries experience was assessed using the guidelines to standardise the collection of epidemiological data throughout the UK. RESULTS: Children studied in 1997 compared with 1993 were younger, had more decayed but fewer filled teeth, were more dentally anxious, more likely to present in pain and came from families who were in receipt of Government benefits. A linear regression analysis showed that the children with previous experience of DGA had higher levels of dental caries, fewer filled teeth, were older and attended in 1997. An individual regression for 1993 showed that an additional predictor was child attendance pattern. CONCLUSIONS: These findings confirmed the clinical observations and suggested that children who attend in pain and present for repeated DGA represent a group of children with special dental healthcare needs. There is a need for an integrated dental healthcare service to identify such children and provide appropriate dental care facilities for them. PMID- 11405018 TI - Where do VT trainers go? AB - Vocational training (VT) has been running since 1985 in the Northern Deanery, first as a voluntary scheme and continuing right through to the present time. In 1985 there were six vocational practices and now there are 48 one-year VT practices and 12 practices dedicated to pilot the two-year general professional training schemes. During that time many practices have been involved, but not all have reapplied in subsequent years. As it is such a large scheme it is necessary for practitioners to apply every year to be vocational trainers, either as new applicants or those who have had vocational trainees before. This paper looks at the reasons why previous trainers have not reapplied, and looks at any outcomes that may be put in place in order to facilitate or improve the quality and number of VT applications each year. PMID- 11405019 TI - Revisiting the use of glucocorticosteroids in dentistry. PMID- 11405020 TI - Immediate placement and provisionalization of maxillary anterior single implants: a surgical and prosthodontic rationale. AB - Maintaining the existing gingival architecture of a failing maxillary anterior tooth is essential in achieving an optimal aesthetic result. Immediate implant placement has been advocated to minimize tissue loss following extraction. The incorporation of a customized temporary abutment and immediate fixed provisionalization facilitates maintenance of the gingival architecture for optimal aesthetics and eliminates the need for a removable provisional prosthesis during the healing phase. This article describes the surgical and prosthodontic approach of this procedure as well as its clinical rationale. PMID- 11405021 TI - Management of soft and hard tissue surrounding dental implants: aesthetic principles. PMID- 11405022 TI - Sinus grafting using platelet-rich plasma--initial case presentation. AB - Recent research has indicated that an element of the proteins contained in platelet-rich plasma can be utilized to increase the speed of bone deposition and quality of bone regeneration when augmenting edentulous sites for implant placement. The growth factors present in platelet-rich plasma are believed to initiate a sequence of microscopic events that ultimately result in the formation of osseous tissue capable of supporting implants for subsequent prosthetic restoration. This article demonstrates a surgical protocol used to harvest platelet-rich plasma and the manner in which it can be implemented in a clinical procedure for the restoration of partially edentulous patients. PMID- 11405023 TI - Restoration of endodontically treated teeth utilizing cast posts and cores. PMID- 11405024 TI - Facial analysis: a comprehensive approach to treatment planning in aesthetic dentistry. AB - Patient aesthetic expectations and smile enhancement can be achieved through the use of facial analysis. This process allows each member of the restorative team (i.e., clinician, specialist, dental technician) to diagnose the patient and develop a comprehensive treatment plan for his or her specific needs. Treatment planning according to facial architecture and dental configuration allows function and harmonious aesthetics to be improved. This article demonstrates a predictable means to evaluate the components of an attractive face for use as a guide during aesthetic dental treatment. PMID- 11405025 TI - Aesthetic posts and cores for metal-free restoration of endodontically treated teeth. AB - Utilization of contemporary post and core systems has facilitated the aesthetic restoration of endodontically treated teeth. Light transmission and biocompatibility have been enhanced by the introduction of metal-free post systems. The periodontal and endodontic status, root length, and histological structure of the devitalized teeth must be considered in order to achieve successful restoration following endodontic treatment. This article presents various restorative criteria for the aesthetic placement and buildup of post and core materials, as well as the preservation of maximum coronal and root structure. PMID- 11405026 TI - Advances in operative dentistry and fixed prosthodontics. AB - The rapid developments in biomaterials has led not only to improved materials but also to the development of clinical techniques made possible by these advances. Adhesive dentistry remains one of the fastest changing fields and this will most likely continue well into the next decade. Patients' aesthetic awareness and, to some extent, concern about amalgam has produced a growing demand for posterior tooth-coloured restorations. Originally this was met by ceramo-metal crowns and subsequently by the development of posterior composite resins. Recent development of stronger ceramics and better luting have now made all-ceramic restorations possible in posterior sites. Several new techniques for cavity preparation are emerging as alternatives to the conventional handpiece while the quest for amalgam alternatives continues. As with all recent developments there are, by definition, few long-term clinical data yet available to allow a comprehensive evaluation of the relevant materials and techniques. This paper is a summary of the most significant recent advances in fixed prosthodontics and operative dentistry. PMID- 11405027 TI - Clinical effectiveness of mandibular implant-retained overdentures. AB - INTRODUCTION: The literature reports that the treatment of fully edentulous mandibles with implant-retained overdentures has become a recognised form of therapy: however long-term data on the success are limited. The aim of the present study was to describe the clinical effectiveness of mandibular implant supported/retained overdentures in the management of a group of edentulous patients. MATERIALS AND METHODS: The study group consisted of 59 consecutive edentulous patients attending the clinic of prosthetic dentistry at Cardiff Dental Hospital, who had been provided with implants in the mandible for complete overdenture retention. Several clinical parameters were examined in each patient: mobility of each implant, probing pocket depth, the presence or absence of plaque and calculus on each implant, bleeding index, marginal bone loss, jaw bone quality and quantity, complications that had occurred with the implants and patients opinions of the treatment. DISCUSSION: Of implants placed, 97% remained in function. Three fixtures had been lost. There was a high frequency of bleeding on probing. Complications associated with treatment included damage or looseness of abutment screws, fracture of the bar, entrapment of food beneath the overdenture and looseness of the opposing non-implant-retained complete denture. CONCLUSION: In general a very high degree of patient satisfaction with the implant-retained overdentures was recorded. PMID- 11405028 TI - Private dentistry: why we need the facts. AB - Rumor and speculation abound within the dental profession about practitioners withdrawing from the NHS to deliver more private dentistry. Due to an absence of effective monitoring or research into this issue the real situation is unclear. We decided to find out what proportions of the gross incomes of general dental practitioners in the East Riding Health Authority were generated by private dentistry. We also sought to establish if they perceived any differences between the quality of their private and NHS work. Our findings and the issues raised are considered for general dental practitioners, for people residing in the authority, and for managers and policy makers. We conclude that the effective management of the supply of NHS dentistry should include a method of systematic monitoring of trends in the delivery of private dental services and the impact on the availability of NHS care. Effective measures are also needed to influence the number and location of dentists in health authorities in England and Wales to ensure adequate and equitable access to NHS dentistry. PMID- 11405029 TI - Stress and the GDP. PMID- 11405030 TI - A case-control study of differences between regular and causal adult attenders in general dental practice. AB - OBJECTIVES: To assess whether adults attending a dental practice for regular dental care have better oral health than adults attending casually in response to a dental problem, and to explore the barriers to asymptomatic attendance. METHODS: An observational case-control study comparing the characteristics of 100 regular attenders with 100 causal attenders in one general dental practice in North Staffordshire. All study subjects were aged 18 years or over. Sociodemographic characteristics of the two groups were collated, including age, gender, social class, marital status, employment status and smoking status. The primary outcome measure was the observed number of teeth with dentinal caries diagnosed using bitewing radiographs. RESULTS: Regular attenders were observed to have better oral health with respect to dental caries and tooth mobility (p < 0.05). This was not explained by the observation that causal attenders were more likely than regular attenders to be male, aged 18-44 years, in social class III or IV. Secondary outcomes, including number of subjects with mobile teeth and teeth with > 30% bone-loss, were also significantly worse in the casual attenders. However, the median number of teeth present in both groups was 27. In regular attenders, the most common reason for attending was to 'keep the teeth' (96%). In casual attenders, 'fear/dislike of dental treatment' was the most frequent indicator of non-attendance (56%). CONCLUSIONS: In our study, adults who regularly attended general dental practice were shown to have better oral health, including less overall tooth decay, mobility and bone-loss, compared with adults who did not attend on a regular basis. Assuming this result to be externally valid, a challenge for the dental profession in the future will be to develop effective oral health promotion initiatives. PMID- 11405031 TI - An overview of reasons for the placement and replacement of restorations. AB - AIM: Surveys on reasons for the placement and replacement of restorations have been conducted in various countries. The purpose of this paper is to bring together and review the data of 10 such surveys with similar methodology. METHOD: The studies reviewed are all based on the protocol described by Mjor in 1981. RESULTS: The surveys reviewed provide data on the reasons for the placement and replacement of a total of 32,777 restorations. Notwithstanding differences between countries, different groups of practitioners and minor variations in methodology, the data included in the selected surveys indicate that everyday clinical practice in Scandinavia, UK and USA during the 1980s and 1990s has involved more replacement than initial restorations, with the ratio of initial to replacement restorations ranging from 1:1.1 to 1:2.4 for amalgam and 1:1.1 to 1:3.8 for composite materials. Primary caries has been consistently found to be the principal reason for the provision of initial restorations of amalgam and composite. The principal reason for the replacement of restorations of amalgam and composite has remained secondary caries as diagnosed clinically. Material failures (marginal degradation, discoloration, bulk fracture and loss of anatomic form) accounted for the replacement of more restorations of composite than amalgam. Recent surveys have reported secondary caries as the principal cause of failure of restorations of glass-ionomer cements and related materials. CONCLUSIONS: The clinical diagnoses of caries may continue to be found to account for the placement and replacement of most intracoronal restorations. Surveys of the type reviewed provide valuable insight into patterns of provision of dental care and highlight research priorities in relation to direct restorations. PMID- 11405032 TI - Diagnostic quiz #49. Case no. 1. Proliferative verrucous leukoplakia. PMID- 11405033 TI - Diagnostic quiz #49. Case no. 2. Very poorly differentiated carcinoma. PMID- 11405034 TI - Preparing for a deposition. PMID- 11405035 TI - The real bottleneck in providing quality dental care. PMID- 11405036 TI - The crisis in healthcare. PMID- 11405037 TI - Children's oral health in Texas. PMID- 11405038 TI - Reaching out--TDA dentists strive to meet the access challenge. PMID- 11405039 TI - Looking out for the less fortunate. PMID- 11405040 TI - Pyogenic granuloma associated with aberrant tooth development. PMID- 11405041 TI - Oral and maxillofacial pathology case of the month. Early invasive squamous cell carcinoma. PMID- 11405042 TI - Apnea treatment may cause alterations in adult occlusion. PMID- 11405043 TI - Women's life cycles may affect oral tissues. PMID- 11405044 TI - Alveolar process can indicate bone loss elsewhere in the body. PMID- 11405045 TI - To prescribe or not to prescribe? PMID- 11405046 TI - How to obtain uniformity of occlusal detail. AB - At a recent Primary Dental Care Editorial Board meeting it was suggested that a series of short articles providing practical advice on clinical procedures might well be appreciated by our readers. This, then, is an invitation to our readers to submit material of this type, for publication. The text may be kept brief by the use of illustrations. The following example is the type of communication that we have in mind, but any information of a practical nature about the use of dental products would be carefully considered. We are grateful to the publishers of Fenestra (3M Dental Products) and the authors for permission to republish this piece first published in 1999. PMID- 11405047 TI - Factors affecting dental attendance following treatment for dental anxiety in primary dental care. AB - AIM: To explore factors affecting patients' dental attendance behaviour following referral from a dental anxiety clinic to a general dental practitioner. DESIGN: A four-year follow up of patients who completed a course of treatment for dental anxiety by conducting semi-structured interviews, face-to-face or by telephone, with confirmation of dental attendance from the dental records. SUBJECTS: Forty one patients who had completed a course of treatment for dental anxiety, in a specially designed community clinic. Mean time since last dental visit before treatment was 7.8 years (range 0.5-29). RESULTS: Twenty-three patients were successfully followed up and 11 (47%) were receiving regular asymptomatic dental care. The dental anxiety scores of those who subsequently became asymptomatic attenders were significantly lower at follow-up than the patients who became symptomatic attenders (P = 0.01). Effective dentist-patient communication was a common theme of the interviews. CONCLUSIONS: At four-year follow-up, dental anxiety was substantially lower in those who subsequently became asymptomatic attenders than those who became symptomatic attenders. A positive dentist-patient relationship had developed with the asymptomatic attenders. PMID- 11405048 TI - Advances in dental imaging. AB - The number of dental radiographs taken in the UK has steadily increased over the past 20 years--recently estimating around 18 million taken in the general dental services alone, and dental radiographs now account for nearly 25% of all medical radiographic exposures. Radiographs remain our most useful diagnostic aid. Their strength is in demonstrating hard tissue pathology, which makes radiographs particularly effective in the maxillofacial region. Although well accepted in this capacity, there remain a number of limitations and drawbacks to conventional radiographs which recent developments have begun to overcome. There have been improvements in the scope and capabilities of dental imaging equipment. There has also been a continuing effort to reduce radiation-induced harm by limiting our exposure to it. This has been possible both through the introduction of new methods and protocols for reducing individual radiation exposures and by the creation of guidelines for selecting radiographs more effectively and thereby reducing the total number of radiographs taken. PMID- 11405049 TI - Occlusal caries diagnosis in molar teeth from bitewing and panoramic radiographs. AB - INTRODUCTION: Previous studies have implied that the panoramic radiograph was inferior to the bitewing radiograph for caries diagnosis. However, these clinical studies lacked a method of validation. The aim of this study was to use an electronic caries meter (ECM II, LODE, Groningen, The Netherlands) to validate occlusal caries diagnosis made from bitewing and panoramic radiographs. MATERIALS AND METHOD: Forty-nine Army recruits were examined with the ECM, and had bitewing and panoramic radiographs taken. In total 299 molar occlusal surfaces were available for examination. Seven examiners viewed the bitewing and panoramic radiographs on two separate occasions and assessed each occlusal surface for dentine caries as 1: almost definitely no caries, 2: probably no caries, 3: unsure, 4: caries probably present, and 5: caries almost definitely present. This was repeated on 20% of the radiographs at two further separate sittings. ECM conductance readings greater than 9 were taken to indicate dentine caries. Examiner decisions that caries was probably and definitely considered to be present were taken as positive diagnoses. RESULTS: Bitewing and panoramic radiographs provided sensitivity values of 0.25 and 0.19 and specificity values of 0.93 and 0.97 respectively. ROC analysis indicated no statistically significant difference in diagnostic quality between the bitewing and panoramic radiographs. Intra-examiner reproducibility was found to be poor to moderate (Kappa values for bitewing = 0.31-0.44, panoramic = 0.07-0.54). CONCLUSION: No difference in overall diagnostic performance was found between bitewing and panoramic radiographs for the diagnosis of occlusal dentine caries. PMID- 11405050 TI - The provision of occlusal splints in primary dental care. PMID- 11405051 TI - An evaluation of attitudes of Midlands female dentists to general dental practice: will it impact upon the future delivery of primary dental care? AB - INTRODUCTION: Both anecdotal evidence and research indicate that there is a developing manpower shortage in the delivery of dental care in the UK. With the predicted increase in registered female dentists, it is possible that future changes in work patterns might occur. If they are occurring, it is important to understand the needs of these female dentists in order to maximise efficient delivery of dental care. The aim of the study was to identify any gender differences that might exist in attitudes to the delivery of primary dental care within general dental practice in the Midlands. METHOD: A Likert-style questionnaire was sent to all private and registered NHS GDPs who worked within the areas covered by three Midland health authorities. RESULTS: The results indicated that the clinical practice of female GDPs appears to involve less private work than their male counterparts and they seem less inclined than male GDPs to proactive development of private practise. Females were less likely than males to be principals and also less likely than males to employ other dentists. Female GDP attitudes towards management, net income and stress at work appears to differ to those of their male counterparts. There was little difference in gender attitude towards professional ethics and maintenance of clinical standards. PMID- 11405052 TI - Is the current access to health information helping or hindering effective decision-making for dentists and patients? Guidelines for dental practice. AB - With the introduction of the Internet there has been a growth both in the quantity and accessibility of information to the public concerning health issues. This improved availability of information does not always lead to a more informed public since there is no quality control of information but it can lead to a public that takes a more active role in their own health and thus is involved in shared decision-making. In order to develop a more informed public in the future, systems for quality control of information have been addressed. These systems can range from the present state of uncontrolled information (no quality control) to full-centralised control (censored information). Between these extremes lie two, more appropriate, decentralised filtering approaches--'upstream filtering' (where third parties set quality criteria and evaluate information) and 'downstream filtering' (where data are rated, labelled and weighted according to consumers' criteria). These systems of filtering are discussed along with recommendations for those using the Internet as a source of information. The paper also documents reliable sources of information for the public, highlights the current concepts of shared decision-making, and provides some guidelines for developing an effective decision-making strategy. PMID- 11405053 TI - Ethnic classification in primary dental care and dental health services research: time to pause for thought. AB - 'Ethnicity' is an important concept in dental health services research and in enabling general dental practitioners to gain insight into their patients values and expectations. Since more health services research is being undertaken in primary dental care settings it is becoming an important issue for dental professionals and researchers in primary care to be aware of. Ethnicity is thought to be related to dental health inequalities and access and is often used as a stratifying variable in many dental studies. The meaning and use of the term however differs among researchers and among the public. It is clear that researchers and professionals need to pause for thought when considering what this often bandied about term actually means and the impact of different definitions. This is illustrated using examples from the authors' own research and published papers in the medical and dental literature. There is also much debate about whether ethnicity--however defined--is an important predictor of differences in dental health in itself or is merely a marker for other factors such as social deprivation or the impact of 'place' on dental health. While the jury on this debate is out we suggest guidelines on the reporting of ethnicity should be outlined in the dental literature--perhaps updating those published in 1996 in the British Medical Journal. PMID- 11405054 TI - Tissue microdissection techniques in quantitative genome and gene expression analyses. AB - Current advances in quantitative genome and gene expression analyses allow precise molecular genetic fingerprinting of tumor tissues. A crucial factor for the reliability of the data obtained with these refined techniques is the use of morphologically well-defined cell populations. Microdissection technology has been developed to procure pure cell populations from specific areas of tissue sections under microscopic control. This review covers techniques of tissue microdissection in the context of commonly used methods of quantitative genome and gene expression analysis. The first part of the review will summarize the technical aspects of various methods developed for tissue microdissection. In the latter part, current applications of quantitative genome and gene expression analysis techniques employed in microdissected tissue samples will be described. PMID- 11405055 TI - Differential expression of major gap junction proteins, connexins 26 and 32, in rat mammary glands during pregnancy and lactation. AB - We examined the expression pattern of two major gap junction proteins, connexin 26 (Cx26) and connexin 32 (Cx32), in rat mammary glands during pregnancy and lactation. Immunohistochemically the two different Cxs were coexpressed in acinar cells and were independently modulated according to the physiological cell activity. Western blot analysis demonstrated that Cx26 gradually increased from early pregnancy, while Cx32 rapidly and dramatically increased at 16 h after parturition, and that both Cxs reached a maximum early in lactation. Increased expression of both Cxs was confirmed by Northern blot analysis showing that their mRNA transcripts were significantly induced on the day of parturition. We also analyzed double-immunofluorescent staining for Cx26 and Cx32 on a confocal laser scanning microscope, in order to examine colocalization of these Cxs in situ. Cx26 immunoreactivity mostly overlapped with Cx32-positive sites in acinar cells of lactating mammary glands, indicating that both Cxs were colocalized together in the same gap junctional plaques in lactation. These results suggest that upregulation of Cx26 and Cx32 in acinar cells at lactating stages, with colocalization in the same gap junctional plaques, may be important for control of secretion by acinar cells in rat mammary glands. PMID- 11405056 TI - Effects of ATP on intracellular calcium dynamics of neurons and satellite cells in rat superior cervical ganglia. AB - Adenosine 5'-triphosphate (ATP) which is released from neuronal and non-neuronal tissues interacts with cell surface receptors to produce a broad range of physiological responses. The present study addressed the issue of whether the cells of the superior cervical ganglia (SCG) respond to ATP. To this end, the dynamics of the intracellular calcium ion concentration ([Ca2+]i) of neurons and satellite cells in intact SCG was analyzed by laser scanning confocal microscopy. ATP produced an increase of [Ca2+]i in both neurons and satellite cells; initially, ATP elicited [Ca2+]i increase in satellite cells and, subsequently, a [Ca2+]i change in neurons was observed. P1 purinoceptor agonists had no effect on this process, but P2 purinoceptor agonists induced [Ca2+]i increase and suramin totally inhibited ATP-induced [Ca2+]i dynamics in both neurons and satellite cells. In satellite cells, Ca2+ channel blockers and the removal of extracellular Ca2+, but not thapsigargin pretreatment, abolished ATP-induced [Ca2+]i dynamics. In contrast, thapsigargin pretreatment abolished ATP-induced [Ca2+]i dynamics in neurons. Reactive blue-2 inhibited the ATP-induced reaction on neurons alone. Uridine 5'-triphosphate caused a [Ca2+]i increase in neurons and alpha,beta methylene ATP caused a [Ca2+]i increase in satellite cells. We concluded that neurons respond to extracellular ATP mainly via P2Y purinoceptors and that satellite cells respond via P2X purinoceptors. PMID- 11405057 TI - TO-PRO-3 is an optimal fluorescent dye for nuclear counterstaining in dual-colour FISH on paraffin sections. AB - Confocal laser scanning microscopy (CLSM) is an extensive but reliable tool for assessing the hybridisation signals in fluorescence in situ hybridisation (FISH). Most CLSMs are equipped with an argon-laser and a helium/neon-laser illumination system with excitation wavelengths of 488, 543 and 633 nm. A protocol for an optimal nuclear counterstaining in combination with dual-colour FISH for these laser illumination systems has not been established so far. Here, we determined the suitability of eleven dimeric and monomeric cyanine nucleic acid stains on paraffin sections of breast carcinoma specimens in combination with dual-colour FISH (Her-2/neu and centromere 17) for CLSM application. Strong staining of cell nuclei was observed for TO-PRO-3 and YO-PRO-3, YOYO-1 and propidium iodide (PI), but only TO-PRO-3 showed specific staining of nuclei without any staining of the cytoplasm. A specific emission in exclusively one distinct fluorescence channel was shown for TO-PRO-3 (633 nm excitation) as well as YOYO-1, BO-PRO-1 and Sytox Green (488 nm excitation), evaluated by a CLSM and confirmed by 3-D fluorescence spectra. High stability of fluorescence intensity was shown for the far-red dyes TO-PRO-3, YO-PRO-3, YOYO-3 and Syto-59 as well as YOYO-1 and PI. Only TO-PRO-3 was due to its high specificity and stability suitable for detection of an amplification of the Her-2/neu gene by dual-colour FISH and CLSM evaluation. PMID- 11405058 TI - Strong hyaluronan expression in the full-thickness rat articular cartilage repair tissue. AB - Articular cartilage lesions have a poor capacity to regenerate. In full-depth articular cartilage defects, the repair process involves an ingrowth of mesenchymal cells from the bone marrow to the injured area, and these cells attempt to restore the lesion with cartilage-like repair tissue. In this study, we investigated histologically the distribution of hyaluronan in the rat repair tissue in relation to other glycosaminoglycans. Full-depth lesions were drilled to the weight-bearing region of rat medical femoral condyle. The rats were divided into two groups: intermittent active motion (IAM) and running training (RT) groups. In the RT group, programmed exercise was started 1 week after surgery, while the rats in the IAM group could move freely in their cages. The lesions were investigated 4 and 8 weeks after the surgery. Semiquantitative histological grading showed no significant differences in the repair between the groups. In normal articular cartilage, hyaluronan was stained mainly around chondrocytes. During repair, strong hyaluronan staining was observed in loose mesenchymal tissue, while in the repair area undergoing endochondral ossification, hyaluronan was intensively stained mainly around the hypertrophic chondrocytes. Remarkably strong staining for hyaluronan was noticed in areas of apparent mesenchymal progenitor cell invasion, the areas being simultaneously devoid of staining for keratan sulphate. In conclusion, hyaluronan is strongly expressed in the early cartilage repair tissue, and its staining intensity and distribution shows very sensitively abnormal articular cartilage structure. PMID- 11405059 TI - Postnatal restoration of the mouse urinary bladder urothelium. AB - Mouse urothelium is disrupted just before birth, followed by a postnatal restoration process which includes cell proliferation, death and differentiation. We assessed urothelial proliferation by the expression of proliferating cell nuclear antigen (PCNA), desquamation by electron microscopy, and apoptosis by TUNEL staining and urothelial differentiation by the expression of uroplakins and cytokeratin 20 (CK20) as well as the apical plasma membrane maturation. Our results indicated that urothelial proliferation was high from birth until about the 14th postnatal day. A majority of basal cells and even occasional superficial cells were PCNA positive during the first 5 postnatal days. Cell death occurred during the first 9 postnatal days. Between birth and day 5, single cells underwent apoptosis, whereas between days 6 and 9 cells mainly desquamated. CK20 and uroplakins were expressed in all superficial cells in postnatal urothelium. Their subcellular distribution characteristically changed in accordance with the progressive differentiation of superficial cells. During the urothelial postnatal development, proliferation activity slowly decreases to the proliferatively quiescent urothelium of the adult animal. Apoptosis is present in the first 9 postnatal days and within a few days of this period it appears simultaneously with desquamation. Superficial urothelial cells gradually differentiate, which is reflected in the changeable morphology of the apical plasma membrane. PMID- 11405060 TI - Expression of the urokinase-type plasminogen activator receptor in human articular chondrocytes: association with caveolin and beta 1-integrin. AB - The urokinase-type plasminogen activator (uPA) in concert with other proteolytic enzymes plays a critical role in cartilage degradation during osteoarthritis. Urokinase receptor (uPAR), a glycosyl-phosphatidylinositol-linked glycoprotein present on the cell surface of various cell types such as cancer cells, fibroblasts, synoviocytes, and chondrocytes, is a key regulator of the plasmin mediated pericellular proteolysis. Recently, in arthritic synovial tissue increased uPAR expression has been detected. By immunohistochemical analysis we observed, in addition, enhanced expression of uPAR in chondrocytes of arthritic samples of human cartilage compared to non-arthritic controls. Using in vitro cultured human chondrocytes, we analyzed whether uPAR is associated with structural proteins, which are known to be involved in cell signaling and activation. uPAR in phorbol-12-myristate-13-acetate-stimulated chondrocytes colocalized with caveolin as well as beta 1-integrin, as demonstrated by double immunostaining with specific antibodies. Furthermore, uPAR was present in caveolae-like structures of chondrocytes as detected by immunoelectron microscopy. Finally, both caveolin and beta 1-integrin were coprecipitated with uPAR-specific antibodies from cell extracts suggesting that these proteins may form functional complexes in human chondrocytes. The localization of uPAR in caveolae and its close association with caveolin and beta 1-integrin points to a significance of uPAR-mediated signaling pathways in human chondrocytes. PMID- 11405061 TI - Immunolocalization of CC10 in Clara cells in mouse and human lung. AB - Two antisera, denoted R41 and R42, were raised against a synthetic peptide from the murine Clara cell-specific protein CC10, and one antiserum, denoted R40, was raised against human recombinant uteroglobin, the human homolog of murine CC10. Purified antigen-specific antisera, denoted R40AP, R41AP, and R42AP were prepared using peptide columns. The purified antisera were characterized by dot blots, immunohistochemistry, and immunoblots. Immunohistochemistry of mouse lung showed specific labeling of Clara cells in distal bronchioles by all three antisera. In human lung, the antiuteroglobin antiserum specifically labeled Clara cells, while the anti-mouse peptide antisera had weak crossreactivity and higher background staining. Electron microscopy revealed immunogold labeling of CC10 granules in Clara cells of mouse lung with all antisera. All antisera also labeled a 5-kDa protein on immunoblots of mouse lung homogenates. The surface epithelium of the alveolar air spaces around the distal bronchioles were CC10 positive suggesting a functional activity for CC10 in the lung parenchyma distal to Clara cells. R40AP immunohistochemical staining of sections of normal human lungs and lungs from patients with surfactant protein B deficiency, bronchopneumonia, and idiopathic alveolar proteinosis illustrate the utility of the anti-human CC10 antibody for diagnostic pathology. PMID- 11405062 TI - Catalase immunocytochemistry allows automatic detection of lung type II alveolar cells. AB - In mammalian lung, type II pneumocytes are especially critical in normal alveolar functioning, as they are the major source of surfactant and the progenitors of type I alveolar cells. Moreover, they undergo proliferation and transformation into type I cells in most types of cellular injury, where flattened type I pneumocytes are selectively destroyed. Hyperplasia of alveolar type II cells has also been described in some human chronic lung diseases. In lung, type II pneumocytes and non-ciliated bronchiolar cells are the unique cell types that contain a considerable amount of peroxisomes. Due to the presence of dihydroxyacetone phosphate acyltransferase and non-specific lipid-transfer protein, these organelles have been suggested to be involved in the synthesis and/or transport of the lipid moiety of surfactant. In the present research, the peroxisomal marker enzyme catalase was immunolocalised at the light microscopic level, utilising the avidin-biotin complex method, in lung specimens excised from newborn, adult and aged rats. In all the examined stages the immunoreactivity was so selective for type II pneumocytes it allowed quantitation of these cells by an automated detection system. This was accomplished on specimens from newborn rat lung, in which labelled alveolar cells were counted by a grey level-based procedure and their main morphometric parameters were determined. PMID- 11405063 TI - Taurine levels and localisation in the stratified squamous epithelia. AB - The content and distribution of the amino acid taurine in squamous epithelia were studied using high-performance liquid chromatography and immunohistochemical methods. Quantitative analysis demonstrated that taurine was highly concentrated in the epidermis (5.49 mumol/g fresh tissue in the hairless skin of the hind footpad of the rat), although the values in the isolated stratum corneum were extremely low (< 0.073 mumol/g in the horny layer of the same skin area). No other analysed amino acid (such as glutamate, glutamine, glycine or alanine) showed this specific pattern of distribution. The immunohistochemical study revealed that in the dog and rat epidermis, taurine was present in the keratinocytes of the granular and upper spinous layers. The basal layer, lower spinous layer and stratum corneum were immunonegative. A similar immunostaining pattern was found in the epithelia of the different organs studied: the mouth, tongue and oesophagus of the dog and rat, the rat forestomach and the rat corneal epithelium. Other cell types, such as sebaceous and muscle cells, were immunolabelled. The existence of a circulating pool of taurine in the epidermis (via taurine release from keratinocytes before they reach the horny layer and its uptake by nearby cells) and its possible roles in these cells are discussed. PMID- 11405064 TI - Binding patterns of 51 monoclonal antibodies to peptide and carbohydrate epitopes of the epithelial mucin (MUC1) on tissue sections of adenolymphomas of the parotid (Warthin's tumours): role of epitope masking by glycans. AB - Warthin's tumours provide a unique opportunity to distinguish and compare monoclonal antibodies (mAbs) to the epithelial mucin, MUC1. In this study, we have applied the range of anti-MUC1 antibodies submitted to the ISOBM TD-4 Workshop for this purpose. mAbs and lectins against MUC1-associated carbohydrate epitopes were also included. Among 39 mAbs to peptide epitopes of MUC1, eight distinct types of staining patterns towards the two epithelial cell layers of Warthin's tumours could be observed. A majority of 27 mAbs reacted preferentially (17) or exclusively (10) with columnar cells, whereas 10 mAbs favoured basal cells (1 of them almost exclusively). The observed staining patterns revealed no correlation with the epitopes. However, after carbohydrate-specific periodate oxidation, 33 of the mAbs stained columnar and basal cells equally well, indicating that epitope masking by glycan side chains was in most cases responsible for the different staining patterns. The results demonstrate the profound impact of glycosylation on immunohistochemistry. Among carbohydrate epitopes, sialyl-TF, sialyl-Le(x), sialyl-dimeric Le(x) and Tn were expressed on both columnar and basal cells (the s-TF3 isomer on columnar cells only). The carcinoma-associated Thomsen-Friedenreich epitope was absent. PMID- 11405065 TI - X-linked ocular albinism (Nettleship-Falls): a novel 29-bp deletion in exon 1. Carrier detection by ophthalmic examination and DNA analysis. AB - BACKGROUND: Mutations in the OA1 gene on the short arm of the X chromosome are known to cause X-linked ocular albinism (x1OA) in males. A four-generation family with this disorder, including asymptomatic carrier females, was investigated by molecular analysis of the OA1 gene. METHODS: DNA samples were available from 22 individuals of this family, including 6 affected males and 6 obligate carriers. The nine exons of the OA1 gene were amplified and further analyzed by SSCP and sequencing. RESULTS: A detailed clinical examination of the index patient and two female carriers showed the typical signs of ocular albinism. Visual evoked potential responses showed markedly asymmetrical responses from the two hemispheres in the affected person as well as in the carriers, as a result of misrouting and decussation of optic nerve fibers. Molecular genetic analysis demonstrated a previously undescribed 29-bp deletion at position 225-253 in exon 1 of the OA1 gene, which segregated in the family. CONCLUSION: Clinical examination combined with molecular genetic analysis enhances the potential for a precise diagnosis for persons at risk of x1OA and provide an accurate basis for genetic counseling. PMID- 11405066 TI - An explicit no response instead of time-out in automated visual-field testing. AB - BACKGROUND: To evaluate the effect of response-acquisition technique on psychometric performance in visual-field testing, the conventional one-button yes/time-out method was compared with a two-button yes/no method for responding whether or not the stimulus was detected. There are a number of situations in which the single-button technique leads to ambiguous results. In this study, we thus expected the yes/no method to reduce tendencies towards habituation and automatic responding. Our hypothesis was that the two-button technique could reduce the rate of erroneous responses. METHODS: Luminance-difference sensitivity for bright stimuli (32') on a photopic background was evaluated at 26 locations within the central visual field (30 degrees) using a specially equalised video display unit and a modified 4/2-dB staircase strategy (six reversals, maximum likelihood threshold estimation). Sixty-one ophthalmologically normal subjects (aged 20-30 years) were examined twice with each method. RESULTS: Mean sensitivities with the two-button yes/no method were found to be, on average, 0.13 dB above those measured with the one-button yes/time-out technique--a difference without clinical relevance. Within-subject variability did not differ between the two methods. However, the less intuitive two-button yes/no method had a slightly higher number of false responses in catch trials. CONCLUSION: Compared to the conventional one-button yes/time-out method, the two-button yes/no method in normal young subjects thus showed little difference in mean sensitivities and equivalent within-subject variabilities. Concerning our initial hypothesis, the yes/no method is of somewhat higher complexity and is not able to reduce the rate of erroneous responses. The one-button yes/time-out method fared a little better in error rate. In summary, the yes/no method is an alternative and additional possibility of response acquisition in visual-field testing, which is worthy of being tested in a clinical study with elderly subjects. PMID- 11405067 TI - Florid diabetic retinopathy (FDR): a long-term follow-up study. AB - PURPOSE: To evaluate efficacy of laser photocoagulation and pars-plana vitrectomy in patients with FDR. METHODS: Eighty-eight patients (155 eyes) with FDR were retrospectively evaluated (mean age 27 +/- 6.0 years; proportion of females 66%; insulin-dependent diabetes 100%; mean duration of diabetes 16.5 +/- 5.8 years; poor metabolic control 83%). The eyes were divided in two groups: group I: 136 eyes amenable to laser photocoagulation and, when necessary, to vitrectomy afterwards (45/136); group II: 19 eyes subjected directly to vitrectomy. RESULTS: In group I (mean follow-up 54.2 +/- 38.7 months) the initial visual acuity (IVA) was 0.61 +/- 0.30 and the final visual acuity (FVA) was 0.47 +/- 0.34; in the 45 vitrectomized eyes IVA was 0.15 +/- 0.24 and FVA was 0.19 +/- 0.25. FDR regressed in 75% and worsened in 25% of the cases. In group II (mean follow-up 46.4 +/- 36.3 months) IVA was 0.1 +/- 0.14 and FVA 0.14 +/- 0.22. FDR regressed in 32% and worsened in 68% of cases. CONCLUSIONS: FDR remains a cause of severe visual impairment in diabetics. Patients at risk of FDR are young females with long standing, poorly controlled insulin-dependent diabetes. Panretinal laser photocoagulation prior to vitrectomy is beneficial; information on this severe form of retinopathy is essential to ensure prompt diagnosis and improve its unfavorable clinical course. PMID- 11405068 TI - Role of inhibitors of isoprenylation in proliferation, phenotype and apoptosis of human retinal pigment epithelium. AB - BACKGROUND: Isoprenoid biosynthesis is known to be essential for diverse cellular functions, including cell proliferation. The aim of this work was to study the effects caused by the addition of different inhibitors of isoprenylation (lovastatin, manumycin A, farnesyltransferase inhibitor III and N-acetyl-S farnesyl-L-cysteine) to human retinal pigment epithelium (RPE) in culture, as potential coadjunctive-to-surgery treatments applicable to proliferative vitreoretinopathy. METHODS: Human RPE cell cultures were established from adult corneal donors. Proliferation levels were evaluated using the incorporation of 5 bromo-2'-deoxyuridine into the DNA. Cell viability was measured by tetrazolium bromide transformation. Apoptosis was determined by DNA fragmentation assay, TdT mediated d-UTP-X nick-end labeling (TUNEL) and phosphatidylserine exposure assessment. Changes in cell morphology and actin cytoskeleton were evaluated using a phase-contrast microscope and by fluorescent staining of actin cables with TRITC-phalloidin. RESULTS: We found that lovastatin showed an important antiproliferative effect on human RPE cells in culture. This effect was clearly dose-dependent, and adding mevalonate could reverse it. We also found that lovastatin induced changes in the distribution of actin cytoskeleton and, finally, that it also induced RPE apoptosis. Manumycin A, farnesyltransferase inhibitor III and N-acetyl-S-farnesyl-L-cysteine also showed antiproliferative effects in RPE. However, they do not have any effect on cell morphology or induction of apoptosis. DISCUSSION: We identified various effects of lovastatin on human RPE cultures: inhibition of cell proliferation, modifications of the phenotype and induction of apoptosis. Interestingly, the addition of different inhibitors of protein isoprenylation only affected the proliferation of the cells. There was no evidence that isoprenylated proteins inhibition is related to lovastatin-induced RPE apoptosis. PMID- 11405069 TI - Transforming growth factor beta 2 levels in the aqueous humor in different types of glaucoma and the relation to filtering bleb development. AB - PURPOSE: To investigate the transforming growth factor beta 2 (TGF-beta 2) levels and total protein levels in the aqueous humor of eyes with different types of glaucoma [primary open-angle glaucoma (POAG), pseudoexfoliation glaucoma (PSX), juvenile glaucoma (JG)], and the relation to filtering bleb development after trabeculectomy. METHODS: Aqueous humor was collected at the beginning of surgery from 52 eyes with glaucoma (29 POAG eyes, 17 PSX eyes, 6 JG eyes) and from 29 control eyes that underwent cataract operation. TGF-beta 2 levels (intrinsically activated and total TGF-beta 2) using ELISA methods as well as total protein concentrations of the aqueous humor were determined. All preoperative clinical data of the glaucoma eyes (age, gender, IOP, previous treatment, type of surgery) were compared with the TGF-beta 2 levels. In 40 of these eyes, the postoperative follow-up (filtering bleb development, need for intervention, IOP) was correlated to the preoperatively determined TGF-beta 2 levels. RESULTS: TGF-beta 2 levels were increased in nearly half of the eyes with POAG and in most of the eyes with JG, but in eyes with PSX, TGF-beta 2 levels were within the normal range. No correlation between TGF-beta 2 levels and age, gender, IOP, previous treatment, or type of surgery, or between TGF-beta 2 levels and protein levels in aqueous humor, was found. Correlation between bleb formation and TGF-beta 2 levels revealed that all but two of the POAG eyes with good clinical outcome (type 1 bleb) had normal levels of activated TGF-beta 2. Of the 13 eyes that needed postoperative intervention (type 2 and type 3 bleb), 8 had high and 5 had normal TGF-beta 2 levels. CONCLUSIONS: PSX eyes differ from POAG and JG eyes not only by their clinical or biomicroscopic appearance, but also by their normal TGF-beta 2 levels in aqueous humor. The fact that most of the POAG eyes with favorable bleb development had normal TGF-beta 2 levels indicated that there might be some relationship between bleb formation and TGF-beta 2 levels. On the other hand, the fact that eyes with less favorable bleb development had both low and high TGF beta 2 levels indicated that other factors are also involved in the scarring of the filtration bleb. PMID- 11405070 TI - Intravitreal acetylsalicylic acid in silicone oil: pharmacokinetics and evaluation of its safety by ERG and histology. AB - BACKGROUND: A new method of intravitreal drug delivery of acetylsalicyclic acid (AS) in silicone oil was investigated for safety and for its pharmacokinetics in the posterior pole of the eye. METHODS: The AS was mixed in silicone oil to a concentration of 1.67 mg/ml. After vitrectomy, 15 NZW rabbits received an intravitreal injection of AS/silicone oil suspension. Clinical examination, pre- and postoperative electroretinography (ERG) and histology were performed. The pharmacokinetics of the distribution of salicylic acid was determined by HPLC analysis at 6 h, 24 h and 5 days in optic nerve, retina, choroid, vitreous, and blood. RESULTS: Clinical examination and histology revealed no adverse effects or signs of toxicity. The ERGs showed no significant difference between the pre- and postoperative results. The salicylic acid concentrations demonstrated peak values in the residual vitreous (640.0 micrograms/ml), choroid (446.0 ng/mg) and retina (281.3 ng/mg) at 6 h. At 24 h, the salicylic acid concentration decreased to 20.9 micrograms/ml in the residual vitreous and to 38.5 ng/mg in the retina. At 5 days the retinal level was still 48.1 ng/mg. CONCLUSIONS: AS delivery by intravitreal administration of loaded silicone oil is a safe method and results in high concentrations of salicylic acid in the posterior segment of the eye while maintaining low blood levels. PMID- 11405071 TI - Nitric oxide inhibits the release of acetylcholine in the isolated retina. AB - BACKGROUND: Recent studies have revealed that administration of nitric oxide (NO) donors increases the release of neurotransmitters in various brain regions. In the retina, NO synthetase (NOS) is found in retinal amacrine and ganglion cells, and it is evident that NO is involved in encoding visual information. In the present study, therefore, NO donors were used to study the effect of exogenous NO on the high K(+)-evoked release of endogenous acetylcholine (ACh) in the rat retina. METHODS: Isolated rat retinal preparations were superfused with modified Krebs-Ringer bicarbonate buffer solution. In each experiment, stimulation for 10 min with 30 mM KCl was done twice. The amounts of ACh released by the first or second KCl stimulation were termed S1 and S2 respectively. Test agents were applied just before the second KCl stimulation. The effects of test agents were evaluated as S2 divided by S1. ACh was converted to hydrogen peroxide and electrochemically assayed by high-performance liquid chromatography. RESULTS: S Nitro-N-acetyl-DL-penicillamine (SNAP), an NO donor, dose-dependently inhibited the high K(+)-evoked release of endogenous ACh. Such inhibition by NO was confirmed also by another NO donor, (+/-)-(E)-4-ethyl-2-[(E)-hydroxy imino]-5 nitro-3-hexenamide (NOR3). The inhibitory effect of SNAP was abolished by both carboxy-PTIO, an NO scavenger, and bicuculline, an antagonist of GABAA receptors. CONCLUSIONS: The NO-induced decrease of ACh release is probably due to an NO induced increase of GABAergic system inhibition. PMID- 11405072 TI - Anti-oxidative capacity of various artificial tear preparations. AB - BACKGROUND: Increased UV radiation and ozone exposure may cause "dry eyes of environmental origin", if the normal anti-oxidative capacity of the tear film can no longer cope with the oxidative stress. The use of artificial tears with an adequate anti-oxidative effect may be beneficial in the treatment of dry eyes caused by environmental factors. METHODS: The anti-oxidative capacity of various commercial artificial tear preparations was determined with a modified TRAP procedure. The two preparations with the strongest anti-oxidative effect were then examined for their protective effects against UV or ozone exposure in a hyaluronate model. RESULTS: Of 19 artificial tear preparations tested, only 6 showed strong to moderate anti-oxidative effects. All others were at best weakly anti-oxidative or had no anti-oxidative effect at all. Some of them even acted as oxidants. Although the two most strongly anti-oxidative preparations performed somewhat differently on UV and ozone exposure, they were both found to be highly protective against these important oxidative stress factors. CONCLUSIONS: The anti-oxidative capacity of artificial tear preparations varies widely. While some are strong anti-oxidants, others are less active or even act as oxidants. If the carefully elicited history of patients with dry eyes suggests that noxious environmental factors may be causally involved, artificial tears which are not just lubricants or contain wetting agents, but act as anti-oxidants, should be chosen for treatment from the many commercially available preparations. Such an etiology-oriented concept would probably improve the success rate of treatment for dry eyes. PMID- 11405073 TI - Exudative retinal detachment in macular hole surgery using platelet concentrates- a case report. AB - INTRODUCTION: Retinal detachment after macular hole surgery is a rare complication, usually occurring because of small, peripheral holes. We present a patient with a high bullous exudative retinal detachment following pars plana vitrectomy. CASE REPORT: A healthy 69-year-old patient presented with a macular hole stage III of the left eye. Corrected visual acuity was 20/200. Pars plana vitrectomy was performed without peeling of the internal limiting membrane or an epiretinal membrane, a few drops of platelet concentrate were instilled onto the hole, and the bulbus was filled subtotally with a non-expanding SF6/air mixture. On the 3rd postoperative day a small retinal detachment of the inferior half of the retina was noticed that increased over the next 3 days until it reached the inferior vascular arcade. During the following 3 days a spontaneous remission occurred with complete reattachment of the retina. Six weeks after operation the retina was completely reattached, the macular hole was closed, and the visual acuity was 20/200 with a slight cataract. CONCLUSION: Retinal detachments after macular hole surgery are not always of rhegmatogenous nature but may also be exudative and related to an inflammatory reaction caused by adjuvants. When a retinal detachment occurs immediately after macular hole surgery without detectable holes it may be advisable to wait for some days before reoperation. PMID- 11405074 TI - Epidemiology of basal cell carcinoma of the eyelid in south-western Finland. AB - BACKGROUND: Basal cell carcinoma (BCC) is the most common malignant tumour of the skin in Caucasians. It often appears on the eyelids but there are no population based studies on the incidence of BCC of the eyelid in Nordic countries. METHODS: The mean annual incidence rates of BCC of the eyelid in the region of South Western Finland from 1977 to 1997 were calculated. RESULTS: The mean annual incidence (per 100,000 population) of BCC of the eyelid showed a statistically significant increase from 0.82 (in 1977-1979) to 2.88 (in 1995-1997). The incidence of BCC of the eyelid was 0.8 in the age group of 30-49 years and rose significantly with age (P < 0.0005) to the maximum of 17.2 in the age group of 80 89 years. There were no statistically significant differences in the incidence rates of BCC of eyelid between males and females or between urban, semi-urban and rural populations. CONCLUSION: In South-Western Finland the incidence of BCC of the eyelids more than tripled during the years from 1977 to 1997. This may be due to the increase of the elderly population, because the incidence showed a significant increase with age. PMID- 11405075 TI - Accumulation of latent transforming growth factor-beta binding protein-1 and TGF beta 1 in extracellular matrix of filtering bleb and of cultured human subconjunctival fibroblasts. AB - PURPOSE: To examine immunohistochemically whether extracellular matrix (ECM) of the filtering bleb and of cultured human subconjunctival fibroblasts contains latent TGF beta binding protein-1 (LTBP-1) and TGF beta. METHODS: An enucleated human eye that had undergone trabeculectomy and cultured human subconjunctival fibroblasts were processed for light microscopic immunohistochemistry. Antibodies against LTBP-1, collagen types, fibrillin-1 and TGF beta s were used. TGF beta 1 was located by detecting beta 1-latency associated peptide (LAP). RESULTS: LTBP 1, beta 1-LAP and fibrillin-1 were all located in the subepithelial ECM as well as in the basal epithelial cells of the conjunctiva over the filtering bleb. TGF beta 2 and beta 3 were immunolocated to epithelium and/or fibroblasts/keratocytes. ECM deposited in confluent fibroblast cultures was positive for beta 1-LAP, LTBP-1 and fibrillin-1, whereas sparse cells were negative. CONCLUSIONS: LTBP-1, beta 1-LAP and fibrillin-1 are co-localized to the ECM of the filtering bleb and of cultured conjunctival fibroblasts. Both conjunctival epithelium and fibroblasts are considered to be the source of TGF beta in healing bleb. ECM secreted by in vivo and in vitro subconjunctival fibroblasts may works as a scavenger or repository of TGF beta. PMID- 11405076 TI - Location of lesions in multiple evanescent white dot syndrome and the cause of the hypofluorescent spots observed by indocyanine green angiography. AB - PURPOSE: To determine the location of the lesions in the retina of a patient with multiple evanescent white dot syndrome (MEWDS) and to resolve the conflict in the cause of the hypofluorescent spots observed in the late phase of indocyanine green angiography (ICGA). CASE REPORT: A 27-year-old woman presented with a unilateral enlarged blind spot and a central scotoma. Fundus examination of the left eye showed many round, indistinctly circumscribed white dots extending from the posterior pole toward the periphery. Fluorescein angiography demonstrated hyperfluorescence due to staining at the location of the white dots. Staining was also observed on the venous wall which was consistent with periphlebitis. The location of the scotomas corresponded with the hypofluorescent spots observed around the optic disc and in the macula in the late phase of ICGA. The scotomas disappeared with the resolution of the hypofluorescent spots, and the sites of other hypofluorescent spots were in accord with the sites of periphlebitis. Visual evoked cortical potentials disclosed no laterality, and the critical fusion frequency was reduced but normalized with the disappearance of the scotoma. CONCLUSION: The initial lesions of MEWDS occur in the retinal pigment epithelium and photoreceptor cells, but when MEWDS is complicated by periphlebitis, the inflammatory lesions extend to the inner layers of the retina. The inflammatory changes involve the choroid and all layers of the retina, which then block the weak background fluorescence in the later phase of ICGA and cause the hypofluorescent spots. The visual field defects are probably caused by retinal dysfunction due to the inflammatory changes. PMID- 11405077 TI - On being responsible: ethical issues in appeals to personal responsibility in health campaigns. AB - Appeals to personal responsibility are highly prevalent in health communication campaigns, but their use entails both moral and strategic considerations. This article provides an overview of the notion of personal responsibility as a persuasive appeal in public health communication campaigns and an analysis of concomitant ethical implications. Whereas the issue of responsibility often is acknowledged by practitioners and scholars as a perennial challenge in health interventions, conceptual tools for the identification of its subtle manifestations are not readily available. This article outlines a framework that contextualizes potentially paradoxical consequences of campaign appeals to personal responsibility that can be explained by the medieval allegory of the "Tragedy of the Commons," psychological attribution theory, and public health concerns regarding "blaming the victim." Practice-oriented questions are introduced to help identify ethical issues in personal responsibility appeals that can be utilized in the design and implementation of health campaigns. PMID- 11405078 TI - Personal communication networks and the effects of an entertainment-education radio soap opera in Tanzania. AB - The personal networks of listeners and nonlisteners to an entertainment-education radio soap opera in Tanzania are examined to determine the effects of interpersonal discussion of the soap opera's educational themes of family planning and HIV prevention. Listeners are more likely to discuss these two educational issues in their personal communication networks and are also more likely to have other listeners to the radio program in their personal communication networks. Respondents demonstrate a relatively high degree of homophily with their network partners and are more likely to discuss matters arising from the radio program with their network partners who are of similar tribal membership, religious affiliation, and gender, and those who are equally or more highly educated than themselves. PMID- 11405079 TI - Understanding barriers to preventive health actions for occupational noise induced hearing loss. AB - A theoretically based formative evaluation was conducted with coal miners in the Appalachian Mountains who were at high risk for noise-induced hearing loss (NIHL). The results of four focus groups indicate that despite high levels of knowledge, strong perceived severity of negative consequences, and strong perceived susceptibility to hearing loss, two main categories of barriers (environmental and individual) keep coal miners from using their hearing protection devices (HPD). Further analysis suggests that the environmental factors, rather than individual variables, more strongly influence decisions against protective actions. Recommendations and practical implications are offered. PMID- 11405080 TI - The reporting of crime and violence in the Los Angeles Times: is there a public health perspective? AB - The authors examine how crime and violence is reported and framed by the Los Angeles Times. Using a public health perspective, we examine whether health oriented variables, such as causal factors and societal effects of crimes, are present in crime news stories. The classic stereotyping of crime and violence framing is strongly present in the Times. We discuss what changes would be useful to provide news consumers with a more accurate picture of crime in their community. PMID- 11405081 TI - Health literacy: making a difference in the USA. PMID- 11405082 TI - A comparison of views of individuals with type 2 diabetes mellitus and diabetes educators about barriers to diet and exercise. AB - Diet and exercise are the cornerstones of treatment for persons with type 2 diabetes mellitus, yet patients find these areas of self-management to be the most difficult. Considerable research has indicated that barriers to diet and exercise are critical influences determining adherence to diet and exercise plans. Standards of practice require educators to assess patient barriers to self management. However, little research has investigated whether patients and educators perceive these barriers similarly. This project's objectives were to compare and contrast patients' and educators' perspectives of patient barriers to following a meal or exercise plan, and to identify differences in patients' perceived barriers as related to patient characteristics. Patients with type 2 diabetes mellitus (n = 97) from three eastern Washington area hospitals and diabetes educators (n = 143) from the Washington Association of Diabetes Educators (WADE) were recruited for a mail survey. From the patient survey, the proportion of patients on meal plan (52%) or exercise plan (26%) was low. Certain barriers were prominent for both patients and educators relative to diet (difficulty maintaining a diet away from home, liking foods not in the meal plan) and exercise (not a high priority, weather). However, multivariate analyses indicated that patients and educators view barriers differently. Similarities and differences between educators and patients in response to barriers are discussed relative to enhancing diabetes counseling and education, and overall communication between educators and patients. PMID- 11405083 TI - Genesis and pathophysiology of lower gastrointestinal bleeding. AB - The incidence of lower gastrointestinal bleeding (LGB) is estimated to be 20-30 per 100,000 of the adult population at risk, which is clearly correlated with increasing age. The problem of LGB is identification of the bleeding source. LGB stops spontaneously in 80% of cases, but 10% of bleeding sources cannot be identified, and rebleeding occurs in 25%. The quality of LGB--hematochezia, melena, or occult bleeding--may point to the region of the bleeding source, as patient age is correlated with specific pathologies. In many patients, LGB is a leading symptom of a chronic disorder. Most acute peranal bleeding arises from the colon (80%) with colonic diverticula and angiodysplastic lesions as the most frequent reasons. In 5% of cases, LGB is caused by disorders of the small bowel, in most cases due to small-bowel tumors or to Meckel's diverticulum in younger patients. In 15-20%, acute peranal bleeding is caused by lesions in the upper gastrointestinal tract. The intensity of LGB determines the urgency of identification of the bleeding source; however, chronic occult blood loss superimposed by melena may place the patient at risk as early as a patient with hematochezia. Therefore, prompt resuscitation is required in many LGB patients before diagnostic procedures are initiated. PMID- 11405084 TI - Lower gastrointestinal bleeding: therapeutic strategies, surgical techniques and results. AB - Lower gastrointestinal bleeding (LGIB) is normally treated conservatively or by noninvasive techniques. Emergency operations are only necessary when patients with severe hemorrhage cannot be stabilized by emergency endoscopy or angiography. To improve the postoperative outcome it is of importance to operate on the patients without any unnecessary time delay. If the preoperative localization of the bleeding source failed, a total or subtotal colectomy should be regarded as a safe procedure. A blind segmental resection should not be done. Alternatively, several ileotomies or colotomies might be performed in order to localize and treat the bleeding site. Elective surgery is indicated with chronic or recurrent bleeding that cannot be treated conservatively. A meticulous preoperative localization of the bleeding site, including anorectoscopy, endoscopy, angiography and nuclear scan is required. With reliable knowledge of the cause and localization of the suspected hemorrhage, a directed segmental bowel resection should be performed. PMID- 11405085 TI - Early gastric cancer--excellent prognosis after curative resection in 87 patients irrespective of submucosal infiltration, lymph-node metastases or tumor size. AB - BACKGROUND AND AIMS: Despite a decreasing incidence of primary gastric carcinoma over the last decade, the incidence of early gastric cancer has remained unchanged. Some aspects of the surgical treatment (e.g., extent of resection, lymphadenectomy) are still controversially discussed in the literature. PATIENTS/METHODS: Between May 1986 and July 1999, 87 patients were operated upon due to primary early gastric adenocarcinoma. All patients data were analyzed retrospectively. RESULTS: Of 626 patients with primary gastric carcinoma, 87 (13.9%) had an early carcinoma (54 men, 33 women; median age 61 years). In all patients, curative (R0-) gastrectomy could be performed, total in 62 patients (71.4%) and subtotal in 25 patients (28.6%). Postoperative morbidity was 23% and mortality 4.5%. Mucosal tumors were found in 34 (39.1%) and submucosal in 53 (60.9%) patients. Multicentricity was present in eight cases (9.1%). Twelve patients (13.8%) had lymph-node metastases. The 5-year survival rate was 88.8%. The submucosal infiltration, the lymph-node infiltration, the histological type, and the tumor size had no statistically significant impact on prognosis. CONCLUSION: Radical resection of early gastric cancer cured most of the patients, irrespective of lymph-node metastases or tumor size. Multicentricity, increasing incidence of proximal cancers, and low mortality suggest that total gastrectomy may be indicated. Patients with early gastric cancer may benefit from D2 lymphadenectomy, but this has to be assessed in further randomized studies, in particular for those with small mucosal tumors. PMID- 11405086 TI - Protection of the rat liver against rewarming ischemic injury by University of Wisconsin solution. AB - BACKGROUND/AIM: University of Wisconsin (UW) solution has been proven able to prevent liver injury during cold ischemia. During rewarming ischemia, however, the efficacy of this solution in preserving hepatocyte function is unclear. The aim of the present study was to investigate to what extent UW solution protects rat liver during rewarming ischemia. METHODS: Livers were washed out with cool physiologic saline or with UW solution and subjected to rewarming ischemia for periods of 20 min or 45 min followed by reperfusion using a blood-free perfusion model. RESULTS: In comparison with controls, ischemia for 20 min in saline treated livers led to mild depression of hepatocyte function, while UW solution afforded complete protection of the liver. In UW-treated livers, compared with saline-treated livers exposed to ischemia for 45 min, portal flow was slightly but significantly higher, bile production was increased by 62%, and lactate dehydrogenase leakage into the perfusate was reduced by 61%. In an attempt to explain mechanisms of liver protection by UW solution, we found that UW solution inhibited conversion of hypoxanthine into uric acid, but this effect was not associated with decreased degradation of adenine nucleotides in the liver during ischemia. Following 30 min reperfusion, UW solution increased tissue levels of adenosine triphosphate (not significantly) and adenosine diphosphate (significantly). Further, UW solution markedly reduced tumor necrosis factor alpha release by the liver both after ischemia and after reperfusion. CONCLUSIONS: These results create the hypothesis that UW solution may protect liver tissue during ischemia in liver surgery as well as during the implantation stage of liver transplantation. PMID- 11405087 TI - Crohn's disease: bowel resection to protect the proctium in severe perianal disease? AB - Patients with anal Crohn's disease generally have a bad prognosis. Up to 50% end up with an anus praeter or proctectomy. Many of these young people develop psychological and social problems causing them to become invalids. Local surgery and conservative therapy in such aggressive cases of Crohn's disease presenting with recurring perianal fistulae and abscesses are often unsuccessful; the destruction of the proctium continues. The purpose of our study was to look at the outcome of our patients with regard to these aspects. We included all 56 patients suffering from Crohn's disease treated at our clinic from 1984 until 1991 in a retrospective study and focused on the 13 patients with anal manifestation. The mean follow-up was 15.8 years (3-28 years). Often there was no improvement of perianal disease without resection of the involved bowel, especially in cases where both the colon and the rectum were affected. Anal destruction went on. Seven of the 13 patients suffering from anal complications finally received an anus praeter. An previous bowel-resection or the construction of a temporary anus praeter seem to be necessary to protect the proctium irrespective of abdominal symptoms in patients with recurrent severe perianal Crohn's disease. PMID- 11405088 TI - Increased susceptibility to apoptosis in circulating lymphocytes of critically ill patients. AB - BACKGROUND AND AIMS: Lymphocyte apoptosis may influence immune responsiveness in systemic inflammation. Therefore, we investigated whether early signs of apoptosis (i.e., annexin-V binding and cell shrinkage) in peripheral lymphocytes were different among patients with severe sepsis, critically ill, nonseptic patients after major surgery, and healthy individuals. PATIENTS/METHODS: Ten patients with severe sepsis and ten critically ill, nonseptic patients after major surgery admitted to a surgical intensive care unit in a university hospital were included in the study. In addition, ten healthy blood donors were included for comparison. We investigated early signs of apoptosis using flow cytometric measurement of annexin-V binding to the cell surface and cell shrinkage of peripheral lymphocytes. RESULTS: The percentage of apoptotic lymphocytes determined as annexin-V positive and propidium iodide negative cells was increased in freshly prepared cells of patients with severe sepsis (11.4 +/- 0.5%) and critically ill, nonseptic patients after major surgery (18.5 +/- 2.0%) relative to healthy blood donors (4.4 +/- 0.5%) (P < 0.05). No significant difference between patients with severe sepsis and patients after major surgery were found. Annexin-V binding increased significantly after OKT-3 stimulation of lymphocytes in patients with severe sepsis (34.4 +/- 1.6%), patients after major surgery (33.8 +/- 3.4%), and healthy blood donors (21.1 +/- 2.8%). No significant difference among groups was detected following OKT-3 stimulation. Furthermore, freshly isolated peripheral lymphocytes of patients with severe sepsis and critically ill, nonseptic patients after major surgery revealed a significantly higher proportion of cell shrinkage than in healthy blood donors (55.0 +/- 2.2%, 21.5 +/- 2.4% vs 3.6 +/- 0.7%; P < 0.05). CONCLUSION: Circulating lymphocytes of critically ill patients show a high degree of early signs of cellular apoptosis. This may contribute to hyporesponsiveness of immune cells in systemic inflammation. PMID- 11405089 TI - Is there any consensus in diagnostic and operative strategy with respect to medullary thyroid cancer? A questionnaire answered by 73 endocrine surgical units. AB - BACKGROUND: The purpose of this investigation was to analyze the individual diagnostic and operative strategy in the treatment of medullary thyroid carcinoma (MTC) in international specialized centers and to assess whether standard procedures are carried out in practice everywhere. METHODS: A questionnaire concerning diagnosis and treatment of patients with primary, persistent, or recurrent sporadic or familial MTC was sent to 263 members of the International Association of Endocrine Surgeons. RESULTS: Primary treatment of MTC does not show significant differences for patients with sporadic or familial disease (Chi square, n.s.), and standard procedures are performed in only 25-40% of patients. Computed tomography scan is the most common localization procedure in persistent or recurrent disease (52-72%), followed by scintigraphy (43-71%), ultrasonography (41-56%), and magnetic resonance imaging (31-49%). In case of negative localization studies, 68-86% of colleagues do not recommend reoperation. In symptomatic patients with stage-IV tumors, however, 84% of colleagues advocate reoperation to provide relief from the tumor burden. CONCLUSIONS: Even with experienced endocrine surgeons, a consensus to uni- and/or bilateral neck dissection in primary MTC is lacking. The majority of authors supports at least total thyroidectomy with central lymph-node dissection. In recurrent disease, there is a general tendency to reoperate in case of positive localization studies and in case of symptomatic disease. PMID- 11405090 TI - Intra-abdominal hypertension and abdominal compartment syndrome. AB - BACKGROUND: The effects of increased intra-abdominal pressure in various organ systems have been noted over the past century. The concept of abdominal compartment syndrome has gained more attention in both trauma and general surgery in the last decade. This article reviews the current understanding and management of intra-abdominal hypertension and abdominal compartment syndrome. METHODS: Relevant information was gathered from a Medline search of the English literature, previous review and original articles, references cited in papers, and by checking the latest issues of appropriate journals. RESULTS AND CONCLUSION: Akin to compartment syndrome in extremities, the pathophysiological effects of increased intra-abdominal pressure developed well before any clinical evidence of compartment syndrome. These effects include cardiovascular, pulmonary, renal and intracranial derangement, reduction of intestinal and hepatic blood flow, and reduction of abdominal wall compliance. Although abdominal compartment syndrome is more commonly noted in patients with abdominal trauma, it is now evident that non-trauma surgical patients could also develop the condition. Early initiation of treatment for intra-abdominal hypertension is currently advocated in view of the possibility of subclinical progress to the full-blown abdominal compartment syndrome. PMID- 11405091 TI - An alternative approach for the surgical management of hydatid disease of the liver. AB - Hydatid disease of the liver still remains a serious medical problem. As a result of increased travelling and immigration, it has also appeared in previously unaffected countries and is now well manifested in countries where the medical personnel lack the experience to treat this problem. We report an alternative method for the surgical management of hydatid disease of the liver, especially that located over the right superior-posterior aspect of the liver. Through a lateral right thoracotomy, we have a better exposure of the cyst and by using the right hemidiaphragm we achieve the same beneficial effect as omentoplasty does. PMID- 11405092 TI - Classification and surgical treatment of incisional hernia. Results of an experts' meeting. AB - BACKGROUND: The treatment of incisional hernia (IH) is a current problem in modern surgery. Many important aspects of incisional hernia surgery are yet to be answered, especially the choice of surgical technique and its adaptation to the individual patient. The aim of this experts' meeting was to resolve some current questions in incisional hernia surgery and to organise an international hernia register. METHODS: An international panel of ten experts met under the auspices of the European Hernia Society (GREPA) to investigate the classification and therapeutic alternatives for incisional hernia. Prior to the conference, all experts were asked to submit their arguments in the form of published results. All papers received were weighted according to their scientific quality and relevance. The information from this correspondence was used as a basis for panel discussion. The personal experiences of the participants and other aspects of individualised therapy were also considered. RESULTS: The expert panel suggested a new classification of incisional hernia based on localisation, size, recurrences and symptoms. All experts agreed that the fascia duplication and the fascia adaptation should only be used for small incisional hernias. Fascia duplication is of value only in the horizontal direction. The technical details and the pros and cons of each procedure were discussed for prosthetic implantation using onlay and sublay techniques and the technique of autodermal hernioplasty. CONCLUSIONS: The management of incisional hernia is currently not standardised. In order to answer relevant questions of incisional hernia surgery, an international hernia register should be established. PMID- 11405093 TI - Diverticular disease-associated hemorrhage in the elderly. AB - Lower gastrointestinal bleeding is frequent in the elderly secondary to diverticular disease and occurs in about 10-30%. It is the most frequent cause of lower gastrointestinal hemorrhage (about 40% of cases) followed by angiodysplasia (up to 20% of cases). The incidence of both diseases increase with age, but the patient's general condition and state of health decrease. Often cardiovascular morbidity coexists, resulting in an eventual risk of ischemic consequences. The intensity of bleeding varies from massive to occult. In diverticular disease, hemorrhage is caused by rupture or erosion of the vasa recti stretched by diverticula. Classically inflammation is absent. Although most diverticula (> 90%) are located in the sigmoid colon, bleeding originates more frequently from the right (> 50%) than the left colon. The preferred diagnostic tool following resuscitation is colonoscopy with an ability to locate the site of bleeding in up to 90% of cases. Additionally, injections and thermocoagulation are available to control bleeding endoscopically with a success rate of about 27%. Angiography is considerably variable concerning positive results (13.6-86%), has a complication rate of about 10% and is expensive. Hence, it is a second-line diagnostic method. Diverticular hemorrhage will cease spontaneously in about 90% of cases. Therefore, conservative treatment is preferred. Patients with persistent, massive or recurrent bleeding despite active conservative measures require surgical treatment. If surgical intervention is necessary, the site of hemorrhage must be sought to allow segmental resection. However, if the source of blood loss cannot be located, a subtotal colectomy is justified. PMID- 11405094 TI - Characterization of two genes encoding the mitochondrial alternative oxidase in Chlamydomonas reinhardtii. AB - Two cDNA clones (AOX1 and AOX2) and the corresponding genes encoding the alternative oxidases (AOXs) from Chlamydomonas reinhardtii were isolated and sequenced. The cDNAs, AOX1 and AOX2, contained open reading frames (ORFs) encoding putative proteins of 360 amino acids and 347 amino acids, respectively. For each of the ORFs, a potential mitochondrial-targeting sequence was found in the 5'-end regions. In comparison to AOX enzymes from plants and fungi, the predicted amino acid sequences of the ORFs showed their highest degree of identity with proteins from Aspergillus niger (38.1% and 37.2%) and Ajellomyces capsulatus (37% and 34.9%). Several residues supposed either to be Fe ligands or to be involved in the ubiquinol-binding site were fully conserved in both C. reinhardtii putative AOX proteins. In contrast, a cysteine residue conserved in the sequences of all higher plants and probably involved in the regulation of the enzyme activity was missing both from the AOX1 and AOX2 amino acid sequences and from protein sequences from various other microorganisms. The transcriptional expression of the AOX1 and AOX2 genes in wild-type cells and in mutant cells deficient in mitochondrial complex III activity was also investigated. PMID- 11405095 TI - Marker free transgenic plants: engineering the chloroplast genome without the use of antibiotic selection. AB - Chloroplast genetic engineering offers several advantages over nuclear transformation including high levels of gene expression and gene containment. However, a consequence of placing a transgene in the chloroplast genome is that the antibiotic resistance genes used as selectable markers are highly amplified. Engineering genetically modified (GM) crops without the use of antibiotic resistance genes should eliminate the potential risk of their transfer to the environment or gut microbes. Therefore, the betaine aldehyde dehydrogenase (BADH) gene from spinach was used in this study as a selectable marker. The selection process involves conversion of toxic betaine aldehyde (BA) by the chloroplast BADH enzyme to non-toxic glycine betaine, which also serves as an osmoprotectant. Chloroplast transformation efficiency was 25-fold higher in BA selection than with spectinomycin. In addition, rapid regeneration was obtained. Transgenic shoots appeared within 12 days in 80% of leaf disks (up to 23 shoots per disk) under BA selection compared to 45 days in 15% of disks (1 or 2 shoots per disk) under spectinomycin selection. Southern blots confirmed stable integration of foreign genes into all of the chloroplast genomes (approximately 10,000 copies per cell) resulting in homoplasmy. Transgenic tobacco plants showed 15- to 18 fold higher BADH activity at different developmental stages than untransformed controls. Transgenic plants were morphologically indistinguishable from untransformed plants and the introduced trait was inherited stably in the subsequent generation. This is the first report of genetic engineering of the higher plant chloroplast genome without the use of antibiotic selection. The use of naturally occurring genes in spinach for selection, in addition to gene containment, should ease public concerns regarding GM crops. PMID- 11405096 TI - Gender-associated mitochondrial DNA heteroplasmy in the venerid clam Tapes philippinarum (Mollusca Bivalvia). AB - This paper provides evidence of a gender-associated mitochondrial DNA (mtDNA) heteroplasmy in the clam Tapes philippinarum. The pattern of tissue distribution of the two observed mtDNA types (referred to as M and F) parallels that of the doubly uniparental inheritance system of Mytilus. The mitochondrial genetic features of the clam are related to a different rate of evolution of the M- and F type mtDNAs. Since clams are known to be phylogenetically very distant from mussels, the present findings support the hypothesis that the mechanism producing gender-associated heteroplasmy should be considered an ancestral character in the Bivalvia. PMID- 11405097 TI - Ssb1 chaperone is a [PSI+] prion-curing factor. AB - Yeast SUP7' or SUP11 nonsense suppressors have no phenotypic expression in strains deficient in the isopentenylation of A37 in tRNA. Here we show that such strains spontaneously produce cells with a nonsense suppressor phenotype which is related to the cytoplasmically inherited determinant and manifests all the key features of the [PSI+] prion. A screen of a multicopy yeast genomic library for genes that inactivate the [PSI+]-related suppressor phenotype resulted in the isolation of the SSB1 gene. Moreover, we demonstrate that multicopy plasmid encoding the Ssb1 chaperone cures cells of the [PSI+] prion. PMID- 11405098 TI - Contribution of Cat8 and Sip4 to the transcriptional activation of yeast gluconeogenic genes by carbon source-responsive elements. AB - The carbon source-responsive element (CSRE) functions as an activating promoter motif of gluconeogenic genes in Saccharomyces cerevisiae. The positively acting regulatory genes CAT8 and SIP4 encode CSRE-binding proteins which contribute unequally to the regulated expression of a CSRE-dependent reporter gene (85% and 15%, respectively, under conditions of glucose derepression). Deregulated variants of Cat8 and Sip4 are able to bind to the CSRE and allow glucose insensitive gene activation, even in the absence of the other protein, arguing against the physiological significance of heterodimer formation. Gel retardation assays provide evidence for a different binding affinity of Cat8 and Sip4 to at least some CSRE sequence variants. Both efficient biosynthesis of and transcriptional activation by Sip4 require a functional CAT8 gene, while Cat8 was not dependent on SIP4. Thus, our data suggest that the apparent minor importance of Sip4 may be the result of autoregulatory cross-talk among the isofunctional activators Cat8 and Sip4. The derepression deficiency of a CSRE-dependent reporter gene in a strain lacking the Cat1 (Snf1) protein kinase can be suppressed by Sip4 fused to a strong heterologous activation domain. This finding agrees with the idea that phosphorylation by Cat1 may convert Sip4 into a functional activator. PMID- 11405099 TI - Over-expression of the INO2 regulatory gene alters regulation of an INO1-lacZ reporter gene but does not affect regulation of INO1 expression. AB - The yeast INO2 gene encodes a transcriptional activator. Inositol and choline repress transcription of the INO2 gene, and its target genes. That is, INO2 transcription is auto-regulated. This observation prompted two separate investigations to determine if regulation of INO2 is required for regulation of its target genes. One study, using northern blot hybridization, showed that constitutive INO2 transcription did not affect regulation of the INO1 gene, while another study revealed that it severely dampened regulation of an INO1-lacZ gene. By repeating both assays from a single yeast strain we demonstrate that this disparity is due to the different reporter systems. PMID- 11405100 TI - L1-like non-LTR retrotransposons in the yeast Candida albicans. AB - Non-LTR retrotransposons (also known as LINEs) have had a major influence on the structure of many eukaryote genomes. They are abundant in many multicellular eukaryotes, including mammals, but appear to be absent from the yeasts Saccharomyces cerevisiae and Schizosaccharomyces pombe. This absence has, to date, precluded the development of a yeast model system for the study of non-LTR retrotransposons. In this report we describe several families of non-LTR retrotransposons from the yeast Candida albicans. These elements, which we call Zorro elements, are all members of the L1 clade of non-LTR retrotransposons. Some are intact, transcriptionally active and have transposed recently. This finding should allow the development of a yeast model system. PMID- 11405101 TI - Association between variant plasmid formation and senescence in retroplasmid containing strains of Neurospora spp. AB - Serial transfer of Neurospora strains harboring the Mauriceville and Varkud mitochondrial retroplasmids frequently displays erratic growth and senescence. Growth impairment is associated with the formation of variant forms of the retroplasmids that can integrate into the mitochondrial genome, resulting in mtDNA rearrangements and eventual loss of respiratory function. Here, we evaluate the rate at which variant plasmids arise in subcultures of the Mauriceville strain of N. crassa and their association with the senescent phenotype. Although variant plasmid formation preceded senescence, subcultures were found to tolerate variant plasmids for variable lengths of time and no correlation could be made between the specific sequence inserted in the plasmids and the rate or frequency of senescence. In addition, many cultures were found to contain more than one variant plasmid. The lack of concordance between the timing of variant plasmid formation and growth cessation distinguishes these two events, and provides additional insight into the etiology of senescence. We also detected differences in the frequency of senescence between retroplasmid-containing strains of N. crassa and N. intermedia and report the isolation of a strain in which senescence occurs in the absence of variant plasmid formation or detectable alterations in mtDNA. Our findings indicate there are multiple pathways that lead to senescence and suggest there are host-specific mechanisms that suppress the deleterious effects of the variant plasmids. PMID- 11405102 TI - [Pediatric radiology]. PMID- 11405103 TI - [Abdominal magnetic resonance tomography in the child]. AB - MRI provides diagnostic information in multiple abdominal diseases in childhood. Additional information to sonographic findings can be achieved in the diagnosis of abdominal malformation as well as in several inflammatory processes. In childhood cancer imaging MRI is essential at the beginning as well as during therapy to assess response to therapy. Because of radiation protection MRI has to replace CT in abdominal imaging in children. Some technical details have to be considered when children are examined. PMID- 11405104 TI - [Low-field magnetic resonance tomography in pediatric radiology. Possibilities, limitations and prospects]. AB - MRI has been used in pediatric imaging for several years. It provides excellent anatomic detail and tissue characterization combined with the advantage that it is not associated with the application of ionizing radiation to the radiation sensitive infant organism. Low field MRI provides some additional advantages like a lower rate of sedations, easier monitoring of sedated patients and the option of interventional examination and therapy. The disadvantages, however, are the slightly prolonged examination times and the lower signal-to-noise ratio compared to high-field MR scanners. In the future, new techniques using modern gradient echo sequences will provide fast imaging methods with a very high signal-to-noise ratio which could partially replace conventional x-ray imaging. In the presented article we report our experiences in low-field MR imaging of pediatric patients. The possibilities as well as the limitations of this imaging modality are pointed out. PMID- 11405105 TI - [Acute craniocerebral trauma in childhood--early use of MRI]. AB - The purpose of the study was to evaluate the present diagnostic potential of MRI in early stage head trauma und possibly to replace CT studies in children. FLAIR techniques consequently applied as "scout sequences" provided reliable identification of traumatic intra- and extracranial lesions yet during the first measurement in all 24 cases. Follow-up scan confirmed the initial results. The reliability of MRI in acute pediatric head trauma is underlined by the fact that CT scans were no longer necessary within in the last three years. Therefore the imaging algorithm of acute head trauma in children has changed in our institution: medium and high risk patients undergo MRI, in young infants we do US first. HR-CT is reserved for lesions of the visceral cranium. X rays are out. PMID- 11405106 TI - [Acute hematogenous osteomyelitis--exclusion with Turbo-STIR sequence?]. AB - The timely diagnosis and early initiation of antibiotic therapy determine the clinical course of an acute hematogenous osteomyelitis. Consequently, a fast and efficient MRI examination protocol is crucial. We retrospectively evaluated various MR sequences used in the examination of 8 children having osteomyelitis. The examinations were conducted using a 0.5 T MR machine. All patients had a high signal intensity of the lesion in the IR sequence with fat suppression (turbo STIR). An acute osteomyelitis can be excluded in the absence of signal intensity increase in the turbo-STIR sequence without the necessity of having to perform additional sequences. PMID- 11405107 TI - [Quality of ambulatory thoracic radiography in the child--a pilot study]. AB - A quality control of outpatient paediatric chest X-rays was conducted in a sample of patients of one paediatric practice. During a period of eight months the technical image quality was analysed considering both diagnostic aspects and radiation protection. The quality of the 139 examined chest X-rays was inadequate concerning the collimation and focussing of the X-rays and the positioning of the patients. Exposure was estimated as average, sharpness was rated as good. In total 14% of the X-rays were not suitable for medical diagnosis. Image quality of the X-rays of infants (children younger than 6 years) was significantly lower compared to the total sample. Radiation protection standards were not fulfilled. As a conclusion from our results, improvements in outpatient paediatric radiography are urgently necessary. Quality control committees should pay particular attention in radiographs of infants. PMID- 11405108 TI - [Non-neoplastic hypertrophic osteoarthropathy due to infected aortic prosthesis. Diagnostic imaging aspects]. AB - Secondary hypertrophic osteoarthropathy of the legs is a rare complication of arterial graft surgery. The syndrome is composed of arthralgias, clubbing of the toes and periostosis of the long bones and is usually associated with an infected aortic prosthesis. Often a CT-guided biopsy is performed in order to identify the graft as the primary focus. In addition to the lamellar periostosis demonstrable by conventional radiographs, a vast edema of the bone marrow and surrounding soft tissue is disclosed by MRI. Imaging promotes the early surgical reintervention. PMID- 11405109 TI - [Responsibility and ethics in radiology]. PMID- 11405110 TI - ["Rheumatoid" complaints. Osteoarthropathia hypertrophicans (Pierre-Marie Bamberger disease)]. PMID- 11405111 TI - [Sectional diagnosis of orbital tumors. 1: Diagnostic imaging--diseases of the orbits: intraconal compartment]. PMID- 11405112 TI - Effects of growth hormone on growth factors after renal transplantation. AB - Growth retardation occurs frequently in renal transplanted children (RTx) and can be improved by growth hormone (GH) treatment. This study retrospectively examines the insulin-like growth factor-1 (IGF-1) and IGF binding protein (IGFBP) profile of ten growth-retarded children previously given renal allografts, after 1 year of GH treatment period. Ten prepubertal patients (nine boys and one girl) were investigated. They had a mean chronological age (CA) of 11.4 +/- 1.1 years and a mean bone age (BA) of 7.3 +/- 0.9 years. Mean height was -3.9 +/- 0.4 SD units below the mean for CA. The mean body mass index (BMI) was 16.9 +/- 0.6 and the mean inulin clearance was 36.5 +/- 4.9 ml/min/1.73 m2. Recombinant hGH was given at 4 IU/m2/day. Plasma GH, total and free IGF-1, IGFBP-2 and -3 were measured by specific radioimmunoassay (RIA). IGFBPs were characterized by SDS PAGE techniques and ligand and immunoblot analyses. Mean velocity was markedly increased (P < 0.01) after 1 year of GH therapy, expressed as SD score for BA. The range of growth response was wide. The total and free plasma IGF-1 increased (P < 0.01) by about 100% (mean values after GH therapy: 95.9 +/- 2.1 nM and 165 +/- 29 pM, respectively). Plasma IGFBP-3 concentrations increased by about 40% (mean value: 148 +/- 18 pM, P < 0.01), with a concomitant increase in both intact IGFBP-3 and its 30-kDa proteolytic fragment. There was no change in plasma IGFBP-2 concentration. Both mean values of inulin clearance and BMI were unchanged during the treatment. In view of the IGF-1/IGFBP concentration changes, there should have been an even better growth response to GH therapy in these patients. This strongly suggests IGF-1 insensitivity, probably as a result of corticosteroid therapy. PMID- 11405113 TI - The impact of supplemental feeding in young children on dialysis: a report of the North American Pediatric Renal Transplant Cooperative Study. AB - Supplemental feedings are commonly recommended for young children on dialysis but their effect on growth parameters and mortality has not been well documented. We report the results of a North American Pediatric Renal Transplant Cooperative Study (NAPRTCS) survey on the impact of supplemental feedings on growth and mortality in children < 6 years of age at dialysis initiation. Sixty-four nonsurvivors (NonS) were matched with 110 survivors (S) for age at dialysis initiation, primary renal disease, and year of entry into the NAPRTCS database. Questionnaires were completed by participating centers on 137 patients (51 NonS, 86 S). Supplemental feedings were given to 70% of patients and more commonly given to patients < 2 years of age compared to those 2-5 years of age at dialysis initiation (P < 0.001). Supplemental feedings were also more commonly given to patients with nonrenal disease in addition to renal disease compared to those with renal disease only (P < 0.001). In patients receiving supplemental feedings, the method of supplemental feeding was most commonly by nasogastric tube in patients < 2 years of age compared to those 2-5 years of age (P = 0.027). Supplemental feeding use was not different in S compared to NonS. There were no differences in height standard deviation score (SDS), weight SDS, or change in height or weight SDS in patients receiving supplemental feedings compared to those who did not. The height and weight SDS did not improve over time on supplemental feeds. In summary, despite the common use of supplemental feedings in young patients on dialysis, height, weight, and mortality remain unaffected. Prospective long-term evaluation of this therapy is needed to determine the effectiveness of supplemental feeding. PMID- 11405114 TI - Dialysis access induced limb ischemia corrected using quantitative duplex ultrasound. AB - Dialysis access induced limb ischemia (DAILI) is a rare complication after the creation of an arteriovenous fistula in infancy but can cause irreversible ischemic limb damage in severe cases. The incidence of DAILI is higher in bridge graft fistulas than in native fistulas. DAILI patients may be managed by surgically reducing the volume flow in the fistula. However, in the pediatric age group, such a reduction of volume flow may result in thrombosis or an inadequate flow for effective dialysis. Several methods have been described to achieve the delicate balance between essential flow in the fistula and adequate limb perfusion pressure. We have developed a new method employing preoperative duplex ultrasonography to predict quantitatively the reduction in volume flow in the fistula that will allow effective dialysis while providing adequate limb perfusion. The preoperative assessment was reproduced on the operating table using intraoperative duplex. A 3-year-old girl thus treated has had resolution of her ischemic symptoms and maintains long-term patency of her dialysis access. PMID- 11405115 TI - Late-onset downregulation of NaPi-2 in experimental Fanconi syndrome. AB - The pathogenesis of renal phosphate (Pi) leak in Fanconi syndrome is unknown. Disorders of apical membrane transporters, leaky apical membrane, depleted cellular Pi and ATP, and impaired sodium (Na) pumps have been proposed as underlying defects. The present study examined the role of type II Na-Pi cotransport system (NaPi-2) in experimental Fanconi syndrome in rats. Following a single injection of maleic acid (MA), 75 mg/kg body weight i.p., rats were sacrificed after 90 min, 4 h, and 24 h. Renal cortical expression of NaPi-2 mRNA was determined by Northern blotting, and brush border membrane (BBM) NaPi-2 protein by Western blotting. Increased urinary excretion of phosphate was demonstrated as soon as 90 min after MA injection, and was sustained at 4 and 24 h, NaPi-2 mRNA expression and NaPi-2 protein were not decreased after 90 min. NaPi-2 mRNA decreased after 4 h, while NaPi-2 protein decreased only at 24 h. Hence, the immediate phosphaturia in experimental Fanconi syndrome may be independent of NaPi-2 downregulation, possibly resulting from energy depletion or membrane dysfunction. The decrease in NaPi-2 mRNA expression and the subsequent NaPi-2 protein decrease may account for the second-phase phosphaturia. PMID- 11405116 TI - Low birth weight-associated adult hypertension in the rat. AB - Epidemiological surveys have suggested that intrauterine growth retardation is a risk factor for the development of hypertension in later life. A rat model of intrauterine growth retardation, induced by maternal low-protein diet during the second half of pregnancy, was used to study the relationship between birth weight and adult hypertension. The offspring were born at term and were allowed to nurse normally until weaned to standard chow at 4 weeks of age. They had 15% lower birth weights than control offspring, with complete catch-up growth by age 4 weeks. Both females and males developed progressively worsening hypertension beginning at 8 weeks. The 11-month survival rate was 69% versus 100% in control animals. During the early stages of the hypertension, plasma creatinine was normal, plasma sodium concentration was slightly higher than that of control animals, plasma renin activity was suppressed, and the males had mild proteinuria. Renal function remained normal throughout the 11-month observation period, but plasma renin activity gradually rose above control values. Angiotensin-converting enzyme inhibition by enalapril, begun at 8 weeks of age, was effective in completely normalizing the blood pressure, but did not totally prevent the extra mortality. Sprague-Dawley and Wistar rat strains developed equally severe hypertension after maternal protein deprivation, despite their different susceptibilities to nephrosclerosis with aging. In conclusion, maternal low-protein diet resulted in low birth weight and adult hypertension in the rat. Primary sodium retention and expanded extracellular volume may be critical factors during the development of the hypertension. PMID- 11405117 TI - Diffuse crescentic glomerulonephritis in bacterial endocarditis. AB - Renal involvement is common in patients with bacterial endocarditis. The most common bacteria are staphylococci and streptococci, and the commonest renal histopathological lesion is a diffuse proliferative and exudative type of glomerulonephritis. Very rarely, patients may present with an extensive glomerular epithelial crescent formation with a rapid deterioration in the renal function. This study reviews the published literature on diffuse crescentic glomerulonephritis in bacterial endocarditis and reports a 24-year-old male patient with endocarditis due to Capnocytophagia species, a gramnegative facultative anaerobic bacillus, which normally inhabits the oral cavity. Appropriate antibiotic therapy is essential to eradicate the infection. A brief course of corticosteroid therapy may be helpful in those with deteriorating renal function. Plasmapheresis may be useful in those with persistent hypocomplementemia, increased circulating immune complexes, and a progressive deterioration in the renal function. Removal of vegetation or valve replacement may be necessary. Prognosis is generally good. PMID- 11405118 TI - Glomerular extracellular matrix and growth factors in diffuse mesangial sclerosis. AB - Diffuse mesangial sclerosis, isolated (IDMS) or observed in the context of Denys Drash syndrome (DDS) due to WT1 mutation, is characterized by early onset nephrotic syndrome progressing to renal failure. A striking morphological feature is the rapid development of glomerulosclerosis. THE AIMS OF OUR STUDY WERE: (1) to analyze the glomerular distribution of extracellular matrix (ECM) antigens at the early stage of DMS, (2) to determine the composition of the ECM accumulated within the mesangial areas and leading to glomerular sclerosis, and (3) to analyze the expression of growth factors, transforming growth factor-beta 1 (TGF beta 1) and platelet-derived growth factor A (PDGFA). In glomeruli of patients with IDMS and DDS, the glomerular basement membrane (GBM) expression of the heparan sulfate chain of heparan sulfate proteoglycan (HSPG) was decreased from the early stage of DMS, at a time when ECM proteins retained a normal distribution. In fully developed lesions, mesangial and subendothelial accumulation of collagenous and noncollagenous glycoproteins normally expressed in the mesangial area (types IV [alpha 1(IV)2 alpha 2(IV)] and VI collagen, beta 1 laminin, fibronectin, tenascin, and perlecan) increased with progression of mesangial sclerosis. This was associated with mesangial expression of proteins normally restricted to the GBM (agrin, alpha 1/alpha 5, beta 2, and gamma 1 laminin chains) and with accumulation of chondroitin sulfate proteoglycan. The distribution of the alpha 3-alpha 5 chains of type IV collagen was normal. Focal accumulation of types I, III, and V collagen was seen only in severely sclerotic glomeruli. Expression of growth factors TGF beta 1 and PDGFA was increased in 9 of 10 and 5 of 10 patients, respectively. Early decreased GBM expression of the heparan sulfate chain of HSPG could play a role in the proteinuria of DMS patients. Changes in the composition of the ECM accumulated within the mesangial areas are not specific. We speculate that deregulation of the expression of growth factors normally downregulated by WT1, is one of the factors responsible for the rapid and massive mesangial deposition of basement membrane material in DDS. PMID- 11405119 TI - Renal concentrating capacity test using desmopressin at bedtime. AB - A crossover trial was undertaken to evaluate the bedtime administration of desmopressin (Minirin) as a renal concentrating capacity test (RCCT). Medication was given intranasally as a single 20-microgram dose to 58 children ranging from 3 to 15 years of age with suspected or known renal impairment. The night-time test was shown to be a simple and effective means of assessing renal concentrating capacity. Comparison with the standard daytime test resulted in a 60 mosmol/kg higher mean osmolality in the night-time test. The results were reproducible, with a 95% confidence interval of -26 to 43 mosmol/kg. The procedure was easy to perform, with 51 of 52 patients (or their parents) preferring the night-time regimen compared with the daytime test. Night-time desmopressin therefore offers the potential of a user-friendly RCCT in patients with suspected impairment of renal tubular function. PMID- 11405120 TI - Factors affecting renal hydrogen ion excretion capacity in healthy children. AB - In 560 healthy German children and adolescents aged 2.8-22.0 years from the DONALD (Dortmund Nutritional and Anthropometric Longitudinally Designed) study, the relationship between urine pH and renal net acid excretion (mmol/day/1.73 m2) was analysed. A quadratic model showed the best fit (r2 = 0.608). Using logistic regression analysis three parameters (urinary phosphorus excretion, total protein intake and urinary ratio of potassium and sodium) had a significant effect on renal hydrogen ion excretion capacity characterised by the probability of high or low net acid excretion with respect to the urine pH value. Urinary osmolality, in contrast to what has been seen in a previous experimental study with low birth weight infants, along with sex and age had no significant independent effects on renal net acid excretion with respect to the urine pH value over the range of osmolalities observed. In healthy children and adolescents a low fluid intake with high urinary osmolality does not at least substantially decrease the renal capacity of hydrogen ion excretion. PMID- 11405121 TI - Pathophysiology and treatment of IgA nephropathy in children. AB - IgA nephropathy is the most-common primary glomerulonephritis worldwide, and about 20%-50% of patients develop progressive renal failure. The pathogenesis is still unknown and treatment has not yet been established. Knowledge concerning childhood IgA nephropathy has expanded greatly in the last 10 years, and its importance as the major form of glomerulonephritis and major contributor to end stage renal disease is also becoming apparent in children. This review focuses on the pathophysiology and treatment of IgA nephropathy in children. PMID- 11405122 TI - Henoch-Schonlein purpura with antiphospholipid antibodies following an influenza vaccination. PMID- 11405123 TI - Plasma cystatin C values and inulin clearances in premature neonates. PMID- 11405124 TI - [Alternative therapies of local prostatic carcinoma]. PMID- 11405125 TI - [Brachytherapy of local prostatic carcinoma]. AB - Brachytherapy of prostate cancer has become attractive in recent years in Germany. There are several radioactive sources available, which are physically different. Some of them are used as permanent or temporary implants. The permanent sources most frequently used are iodine 125, palladium 103, and gold 198. Iridium is a temporary implant. The techniques used in Germany are low-dose rate (LDR) and high-dose rate brachytherapy, which differ in dose distribution and patient population. The success of prostate cancer brachytherapy depends on patient selection and choosing the right source for the technique used. Best suited for LDR monotherapy is the low-risk patient with a prostate-specific antigen (PSA) level below 10 ng/ml, maximal tumor stage T2b, and a Gleason score of less than 7. PMID- 11405126 TI - [Value of cryotherapy in localized prostatic carcinoma]. AB - Prostate cancer represents the second most frequent malignant male disease in Germany. In the United States, approximately 35,000 men die from prostate cancer annually. The treatment of this disease is of particular interest to both clinical and investigative urologists. Radical prostatectomy and radiotherapy are established standard modalities in the treatment of organ-confined prostate cancer. As in other fields of urology, minimally invasive procedures have gained increased interest in urologic oncology. With cryoablation of the prostate, a minimally invasive therapy for prostate cancer has been available since 1989 and has been used and under investigation since then. Improvements in cryotechnique and progress in transrectal high-resolution ultrasonography enable the surgeon to achieve the curative target of thermoinduced destruction of the whole prostate gland. Control with thermocouples, ultrasound, and double-freeze techniques makes it possible to destroy the whole organ or the region of interest in high-risk patients or in patients who refuse to undergo open surgical procedures. PMID- 11405127 TI - [Therapy of local prostatic carcinoma with high intensity focussed ultrasound (HIFU). Outcome and side-effects]. AB - At the time of diagnosis, prostate cancer is organ confined in 70% of the cases. Of these patients, 25% undergo local therapy (surgery/radiation), and 75% risk disease progression by "watchful waiting" or systemic side effects through hormonal ablation. Local high-intensity focused ultrasound (HIFU) for minimal invasive tissue coagulation (85 degrees C) ablates prostatic tissue with high precision. Follow-up sextant biopsies (1.9) showed 80% of the patients to be cancer free. In those cases with residual cancer, the tumor mass was reduced by more than 90%. The PSA nadir in 97% was < 4 ng/ml, including 61% < 0.5 ng/ml. After primary HIFU, no severe side effects occurred (no fistula, no grade II/III incontinence, no rectal mucosa burn). As auxiliary treatments, all patients received a suprapubic tube (29 days), and 33% needed a transurethral debris resection (TUR 7 g). The patients were released from the hospital within 24 h after treatment. According to the short-term follow-up, transrectal HIFU enables minimal invasive local prostate tissue ablation with high rates of negative biopsies, low PSA nadir, and low complication rate. PMID- 11405128 TI - [Thermoradiotherapy with interstitial thermoseeds in treatment of local prostatic carcinoma. Initial results of a phase II study]. AB - Several experimental data are available regarding the efficacy of the combination of interstitial hyperthermia and conformal radiation therapy. We planned a phase II trial as a member of the special hyperthermia research group of the Deutsche Forschungsgemeinschaft (German Research Society) to determine the efficacy of thermoradiotherapy using interstitial cobalt-palladium thermoseeds in the treatment of patients with localized prostate cancer. Forty-one patients with localized prostate cancer were enrolled in the study between July 1997 and April 2000. The interstitial hyperthermia induced in a magnetic field was applied in six sessions once a week. Conformal three-dimensional radiation therapy was given simultaneously in daily fractions of 1.8 Gy with a total dose of 68.4 Gy. We measured intraprostatic temperatures between 42 degrees and 46 degrees C. No major side effects were observed during the hyperthermia session. The median level of prostate-specific antigen (PSA) decreased from 11.25 ng/ml to 0.88 ng/ml 3 months after treatment and to 0.38 ng/ml 12 months after treatment with a median follow-up of 10 months. The mean prostate volume decreased from 32.6 ml to 26 ml after 3 months of treatment and to 18.5 ml after 12 months. Interstitial hyperthermia is a feasible, well-tolerated procedure in prostate cancer therapy. A significant PSA decrease was observed. Longer follow-up is necessary to determine the efficacy. PMID- 11405129 TI - [Laparoscopic radical prostatectomy. Experiences with 145 interventions]. AB - Encouraged by the groups in Paris, we performed 145 laparoscopic radical prostatectomies between June 1999 and the end of November 2000. The indication for laparoscopic prostatectomy is the same as for open surgery: an organ-confined cancer. Previous abdominal surgery, transurethral resection, and/or relative adiposity are not considered to be contraindications for this laparoscopic procedure. The mean operating time was 255 min; the last 60 procedures took an average of 200 min. In no case was it necessary to convert to open surgery. Worthy of note was the low blood loss of 185 ml on average so that in 98% of the patients no blood transfusion was required. After completing the learning curve, the average indwelling catheter time was only 5.5 days. The postoperative complication rate was 11.7%, consisting mainly of minor complications. Also with regard to continence and potency, the results were representative. Postoperatively, 75%, 86%, 92%, and 93% of the patients were continent after 3, 6, 9, and 12 months, respectively. In our opinion, laparoscopic radical prostatectomy is an alternative to open prostatectomy, offering a number of advantages for the patient and surgeon as well. PMID- 11405130 TI - [Gene therapy and immunotherapy in prostatic carcinoma]. AB - Although local prostate cancer (PC) can be cured in most cases by radical prostatectomy, therapies for metastatic and androgen-independent PC are limited and rather unsatisfactory. Gene and immunotherapy based on progress in molecular biology are novel treatment options especially for these PC stages. In the field of passive immunotherapy, chimeric/recombinant antibodies and derivatives thereof show promising results in early clinical trails (phase I/II). Before treatment, a careful selection of patients who could profit from this therapy is important (theranostics). Concerning active immunotherapy, administration of dendritic cells loaded with PC-specific tumor antigens seems to be an interesting therapy option. Promising gene therapeutic approaches include antisense and suicide gene therapy. Antisense therapy studies revealed the advantage that even systemic treatment does not lead to strong toxic side effects if the target gene is not involved in important cell functions. Improvement of the gene therapy vectors and identification of new therapeutic genes for PC are essential prerequisites for successful application in humans. Present developments of alternative approaches show that future treatments will be very patient specific. PMID- 11405131 TI - [Fluorescence cytology of the urinary bladder]. AB - 5-aminolevulinic acid induced fluorescence cystoscopy is invaluable for diagnosing urinary bladder carcinoma and its precursors. Because neoplastic cells of the urinary bladder possess striking fluorescent properties due to protoporphyrin IX, we initiated a study to evaluate the use of fluorescence microscopy in urinary sediments. In 27 patients suspected of having bladder carcinomas, we instilled 5-aminolevulinic acid into their urinary bladders before transurethral therapy and compared thereafter our studies of standard cytological sediments with those made under fluorescence microscopy. The results of fluorescence cystoscopy and those using urinary sediments for neoplastic cells under fluorescence microscopy correlated extremely well. In this pilot study using fluorescence microscopy, we found that we could diagnose with precision urinary neoplasms of different grades of differentiation. Accordingly, we regard fluorescence microscopy as a valuable complement for standard urinary cytology, especially since with fluorescence microscopy we can readily recognize fluorescing cells of highly differentiated urinary tumors and flat premalignant dysplasias. PMID- 11405132 TI - [Anatomic principles of urinary incontinence]. AB - The morphological fundamentals of urinary continence are still subject to controversy. This was the reason for a renewed examination of the sphincter musculature of the lower urinary tract. This study included 50 male and 15 female autopsy specimens. The organs of the lower urinary tract including the neighboring organs had been removed in their entirety and histologically reprocessed en bloc as a complete series of sections. We were able to demonstrate that the internal sphincter or m. sphincter vesicae is represented as a circular, distinct structure which elliptically embraces the internal urethral orifice. Lamellas of the detrusor are not involved in the formation of the internal sphincter. In females and males, the external sphincter consists of a striated and a smooth muscular part (m. sphincter urethrae transversostriatus et glaber). In transverse sections, the muscle has a horseshoe shape. It is completely separated by connective tissue from the musculature of the pelvic floor. A deep transverse perineal muscle does not exist. The histological findings were used for the construction of a digital three-dimensional model of the anatomy of the lower urinary tract. Computer animations of the model with integrated original histologies were generated and stored as a computer video on a CD-ROM attached to this journal. PMID- 11405133 TI - [Comment on the contribution by W. Dorschner, J. Neuhaus and J.-U. Stolzenburg. Anatomic principles of urinary incontinence]. PMID- 11405135 TI - [Chronic prostatitis--"can we find a way out of the dilemma?"]. PMID- 11405137 TI - [The transfusion law]. PMID- 11405138 TI - [Intermittent catheterization of the urinary bladder--public health requirements and findings of cytotoxicity of selected catheter and catheter lubricant substances in vitro". Comment on the publication by A. Kramer, P. Rudolph, H.-P. Werner and K. J. Klebingat (Hyg Med 2001: 26(1/2):14-24]. PMID- 11405139 TI - [Bacterial colonization of implants in the paranasal sinuses and ear region. What is their role in integration?]. PMID- 11405140 TI - [Blood coagulation tests before tonsillectomy or adenoidectomy?]. PMID- 11405141 TI - [Processing vestibular impulses in the central nervous system. Study based on positron emission tomography]. AB - BACKGROUND AND OBJECTIVE: Functional imaging methods have opened new perspectives for vestibular research. Many authors have investigated the central connections of the system, but the differences between the reports leave further questions open. We investigated the cerebral projection of the vestibular system, using positron emission tomography in right-handed subjects. PATIENTS AND METHODS: Bilateral caloric stimulation was used in every volunteer (n = 6). This can be considered a standard method, which will make it possible to compare the results from different laboratories in the future. A detailed map of activated and deactivated brain regions is included. RESULTS: Changes caused by vestibular stimulation are portrayed. The activated regions partially correspond with previous results in the literature. We would like to point out the Brodmann 6 region as the cortical manifestation of involuntary isometric tightening of muscles. We have found many, previously unidentified regions showing decreased regional cerebral blood flow. CONCLUSIONS: We are the first to point out the functional connection between the hippocampus and the vestibular system in this report. PMID- 11405142 TI - [Diagnosis of questionable mandibular infiltration by squamous epithelial carcinomas. 3-D 99mTc-DPD SPECT reconstruction and (18F) fluoride PET study: diagnostic advantages or unnecessary expense?]. AB - OBJECTIVE: A prospective study was designed to compare computer-aided 3D 99mTc DPD SPECT reconstruction, conventional 99mTc-DPD SPECT investigation (n = 88) and flurine-18 PET (n = 10) in the assessment of mandibular bone invasion by SCC. PATIENTS/METHODS: Between 10/97 and 03/00 88 patients with SCC of the mandibular region were enrolled in this study. In 50 cases mandibular resection (segmental or marginal) was performed basing on the pretherapeutic diagnostic results. RESULTS: No differences could be found between 3D 99mTc-DPD SPECT reconstruction and conventional 99mTc-DPD SPECT investigation. Both techniques revealed the same high sensitivity (100%) and the same specificity of 91.6 (efficiency 95.4%). Flurine-18 PET showed a sensitivity of 100%, but specificity only reached 50% (efficiency 60.0%). CONCLUSIONS: This investigation does not provide evidence that the financial and time consuming techniques of computer-aided 3D 99mTc-DPD SPECT reconstruction and flurine-18 PET show advantages compared to conventional 99mTc-DPD SPECT investigation in the assessment of mandibular invasion by SCC. PMID- 11405143 TI - [Modified uvula flap in therapy of sleep-related breathing disorders]. AB - BACKGROUND/OBJECTIVE: Velopharyngeal incompetence is a well described complication of uvulopalatopharyngoplasty (UPPP) for snoring or obstructive sleep apnea. A new uvulopalatal flap technique (UPF), first presented by Powell in 1996 was now modified by our team. The new technique intends to lateralize the posterior faucial pillars as the UPPP, entirely sparing uvulopalatal muscles. RESULTS: No statistical difference existed in change of AHI before and after surgery in all patients even though 14 patients had a statistically significant decrease in AHI (pre 19.2 +/- 19.6; after 8.2 +/- 9.8; p < 0.05). Snoring decreased clearly (p < 0.05) from 49.4% to 25.9%. PMID- 11405144 TI - [Effect of bioceramics on biological activity of microorganisms]. AB - BACKGROUND AND OBJECTIVE: Biomaterials that come into contact with microorganisms in the middle ear or paranasal sinuses should be tested before clinical application. Thus, it is necessary to test the influence exerted on bacterial growth by biomaterials used as bone substitutes in head and neck surgery before implantation. PATIENTS AND METHODS: In this study, Bioverit, Al2O3 ceramic, and glass carbon were subjected to contamination with typical microorganisms in the middle ear and paranasal sinuses such as Staphylococcus aureus, Streptococcus pneumoniae and salivarius, Escherichia coli, Pseudomonas aeruginosa, Proteus mirabilis and Candida albicans. RESULTS: In the suspension test, all gram negative species were inhibited, but gram-positive microorganisms and Candida albicans were not affected. The Al2O3 ceramic showed the largest inhibition effect while growth inhibition of glass carbon was low. Streptococcus pneumoniae and salivarius can use glass carbon as a nutrient. In contrast, Bioverit and Al2O3 ceramic could not improve the growth of all tested microorganisms. CONCLUSIONS: In conclusion, we think that Bioverit is suitable for implantation in bacterially contaminated regions of the head and neck, whereas glass carbon is unsuitable for this application in reconstructive surgery. PMID- 11405145 TI - [Endonasal temperature and humidity profile after exposure to various climate controlled inspiratory air]. AB - BACKGROUND AND OBJECTIVE: The nose as the first segment of the upper respiratory tract is exposed to different climatic conditions of the ambient air. The purpose of this study was to determine whether nasal air conditioning of ambient air is influenced by a short-term exposure to air of different climatic conditions. PATIENTS AND METHODS: Fifteen healthy volunteers were enclosed into the study. A miniaturised humidity sensor and thermocouple was used for detection of humidity and airway temperature at three detection sites within the anterior nasal cavity. Both parameters were measured at the end of inspiration during breathing of ambient air after exposure to cold dry air, "normal" ambient air, and warm humid air for 10 minutes. RESULTS: The end-inspiratory intranasal humidity during breathing of ambient air increased at all locations within the nose when the temperature and humidity of the exposure condition increased. At the most anterior detection site, the effect of increase in humidity after exposure to the different climatic conditions was the most pronounced. The increase in intranasal temperature had no relation to the different climatic conditions during the short term exposure. CONCLUSIONS: The results suggest that nasal humidification in ambient air can be influenced by short-term exposure to air of extreme temperature and humidity prior to breathing of ambient air. PMID- 11405146 TI - [Genotoxicity of phthalates. On the discussion of plasticizers in children's toys]. AB - BACKGROUND AND OBJECTIVE: Recently, health hazards caused by phthalates, which are added as softeners to plastic materials, have been subject to discussion. The aim of the present study was to measure possible genotoxic impacts on mucosal cells of the upper aerodigestive tract. PATIENTS AND METHODS: Genotoxicity tests for dibutyl phthalate (DBP) and diisobutyl phthalate (DiBP) on human oropharyngeal mucosa in vitro were performed using the alkaline comet assay. Specimens (n = 50) were harvested from the surface of ectomized tonsils. RESULTS: DBP and DiBP caused significant DNA damage in human mucosal cells of the upper aerodigestive tract. The impact of DiBP was higher than that of DBP. CONCLUSIONS: A genotoxic impact of phthalates on human epithelial cells as a hazard to babies and children chewing these materials cannot be excluded and demands further investigation. The DNA damage measured in this study may represent one factor in the complex genesis of neoplasms in the upper aerodigestive tract. PMID- 11405147 TI - [Status of hearing aid use by children in schools for the hearing impaired and deaf]. AB - To evaluate and possibly improve the hearing aid fittings of children attending the Westphalian School for the Hearing Impaired or the Westphalian School for the Deaf, regular pedaudiologic consulting hours were established at both schools. During a 2-year period, 115 children were examined once, 35 children twice, and 5 children three times. The examinations comprised ear microscopy, audiometry, and a check of the hearing aids with a 0.6-cm3 coupler (children up to 7 years) or 2 cm3 coupler, respectively. The following criteria were used to assess the quality of the hearing aid setting: status of the external auditory canal and middle ear, acceptance of wearing the hearing aid, status of the ear mold, technical status of the hearing aid, and its setting. The results were related to four variables: gender, type of school, age, and mean hearing loss. Overall, just 40.9% of all children showed satisfactory hearing aid performance at the first examination and just 37.1% at the second. A significant influence of the variables on the hearing aid performance was documented for hearing loss only. The higher the hearing loss, the more likely the children were to have good hearing aid status. Analysis of the different parameters revealed that an incorrect setting was the main problem, with a rate of 20.9%; the rate of the other parameters varied from 6.1% to 15.7%. Thus, no parameter was of major relevance to the results. The results of the second examination were poorer in most parameters than those of the first. These alarming results, which are probably not only of regional significance, demonstrate that the hearing aid status of children attending schools for the hearing impaired or for the deaf is in urgent need of improvement. PMID- 11405148 TI - [Ductus arteriosus apertus as the cause of recurrent nerve paralysis. A case report]. AB - There are various reasons for paralysis of the vocal folds, which consequently imposes great demands on differential diagnostics. Angiocardiopathies are regarded as very rare cases of etiopathogenesis. In the present case, a persistent arterial duct could be identified as the reason for the paralysis of the left recurrent nerve of a 59-year-old female patient. The necessity of interdisciplinary diagnostics going beyond the field of otorhinolaryngology is emphasized, especially in cases of a paralysis of the recurrent nerve. PMID- 11405149 TI - [Abdominal wall metastases as a complication of percutaneous endoscopic gastrostomy in carcinoma of the upper aerodigestive tract]. AB - Percutaneous endoscopic gastrostomy (PEG) has become an important adjunct in the care of head and neck cancer patients. In the literature of the last 10 years, 16 cases of abdominal wall metastasis after PEG implantation were reported. We performed 387 PEG procedures in patients with head and neck cancers. In this paper, we describe two patients with advanced head and neck cancers who underwent PEG prior to cancer therapy and developed metastatic cancer at the PEG site 3 or 4 months later. Although the mechanism of spread cannot be confirmed, direct seeding from passage through the cancer seems likely. Methods of establishing enteral access which avoid tumor-contaminated fields, such as the use of an overtube during conventional PEG or PEG procedure after tumor resection, may be appropriate in head and neck cancer patients. Another possibility of origin is the hematogenous spread of cancer cells from metastases on the abdominal wall. PMID- 11405150 TI - [Candida mycoses of the mouth cavity]. PMID- 11405151 TI - [Bilateral, progressive vision loss. Aspergilloma of the left sphenoid sinus with concomitant neuritis of both optic nerves]. PMID- 11405152 TI - [Author and medical society legal aspects of falsification of research data]. PMID- 11405153 TI - [DRGs in Germany. Significance for ENT medicine]. PMID- 11405154 TI - [Rehabilitation in chronic facial paralysis. 1]. PMID- 11405155 TI - [Occult cutaneous mastocytosis]. AB - Mastocytosis is diagnosed without difficulty if it presents with easily recognizable lesions of urticaria pigmentosa. Recently, we have identified hardly visible skin lesions of mastocytosis in Hymenoptera venom allergic patients ("occult mastocytosis"). In addition, in approximately 15% of the patients with typical cutaneous lesions, urticaria pigmentosa was at first mistaken for other conditions and thus not linked to simultaneous symptoms of systemic mastocytosis. In most patients with unrecognized mastocytosis, the diagnosis was supported by raised basal serum tryptase levels. Cutaneous mastocytosis is often overlooked and more frequent than assumed. Measurement of basal serum tryptase concentrations can make an important contribution to the diagnosis of mastocytosis, but it does not replace a meticulous skin examination. PMID- 11405156 TI - [Correlation between histopathology and epiluminescent microscopy in nonmelanocytic skin lesions]. AB - Epiluminescence microscopy techniques make surface and subsurface structures accessible to visual examination in vivo. The visualization of a multitude of special morphological features, such as blood vessels, adnexa and deposits of pigments in normal and pathologically changed skin allows a reproducible and topographical stratification. There is a chance of identifying the architecture of the stratum corneum, the epidermis and the papillary bodies. Changes in the amount of melanin can be individually assessed in the stratum corneum and the deeper layers extending into the upper dermis. It is possible to differentiate the orientation, size and color of the blood vessels of the cutaneous vascular plexus, involving the capillaries of the stratum papillare and the subpapillary plexus. PMID- 11405157 TI - [Allergic contact dermatitis in beauty parlor clients]. AB - Occupational contact dermatitis in hair dressers and beauticians has increased in importance in the past years. Type IV-allergies against glyceryl monothioglycate components of permanent waves are most common. Other occupational allergens include bleach components such as ammonium persulfate and hair dye ingredients such as p-phenylenediamine (PPD) and p-toluylene-diamine (PTD) base. Allergies to hair dyes in customers of hair dressers have rarely been observed. Two female patients developed allergic contact dermatitis of the scalp and face after repeated use of Polycolor intensivtonung schwarz and of Movida color. We also review the current literature on type IV-allergies to components of hair dressing products components. PMID- 11405158 TI - [Condylomata acuminata and other HPV-associated manifestations of the genitals and urethra. Guideline of the German STD Society (DSTG)]. PMID- 11405159 TI - [Urethral condylomas. A therapeutic challenge]. AB - BACKGROUND AND OBJECTIVE: Very diverse treatment recommendations exist in the literature on HPV lesions of the urethra. There are no treatment guidelines from the specialist societies. Both these factors have led to a very wide spectrum of heterogeneous treatment strategies in hospitals and medical practices of various specialties. Primary and secondary treatment of urethral condylomata by practitioners as well as a specialized HPV center was evaluated. PATIENTS/METHODS: One hundred and five patients with condylomata of the urethra were studied. Most had been previously treated, often several times, by a variety of specialists. We treated all regardless of prior treatment status with laser therapy. RESULTS: A high percentage of patients treated with different methods in medical practices but also at our high-technology center sometimes showed serious treatment complications and numerous recurrences. Most established methods of treatment for condylomata on the external genitalia are not necessarily applicable to the urethra. CONCLUSIONS: About 20% of urethral condylomata can only be reached by endoscopy. Co-existing urethral malformations as well as complications of therapy are reasons for early cooperation with the urologist or HPV center. PMID- 11405160 TI - [Incidence of fungal involvement in nail psoriasis]. AB - BACKGROUND AND OBJECTIVE: Because it is clinically so similar, onychomycosis is one of the most important differential diagnostic considerations in psoriatic nail changes. We determined the frequency of fungal involvement in nails of psoriatic patients. PATIENTS/METHODS: Fungal colonization of the nails of psoriatic patients with and without clinically obvious nail changes was investigated in a collective of 250 patients compared to a group of 102 non psoriatic persons. RESULTS: There was no significant difference in the frequency of nail colonization with dermatophytes and moulds between healthy persons and psoriatics. Yeasts were more commonly found in psoriatic nails. CONCLUSIONS: Nail changes in psoriatic patients are mostly caused by psoriasis. However secondary colonization by yeasts is more common in psoriatic nails than in the general population. It remains to be clarified why dermatophytes are not more common in psoriatic nails. PMID- 11405161 TI - [UVA1 phototherapy. Pilot study of dose finding in acute exacerbated atopic dermatitis]. AB - BACKGROUND AND OBJECTIVE: UVA1 phototherapy is an new effective treatment modality for acute atopic dermatitis (AD). However there is still some controversy about the optimal UVA1 single and cumulative dose. PATIENTS/METHODS: We compared in a randomized, controlled, prospective pilot study the efficacy of a therapy with 15 treatments of a "high dose" (max. single dose of 130 J/cm2, max. cumulative dose 1840 J/cm2), "medium dose" (max. single dose of 65 J/cm2, max. cumulative dose 975 J/cm2) or "low dose" (max. single dose of 20 J/cm2, max. cumulative dose 300 J/cm2) UVA1 in patients with acutely exacerbated atopic dermatitis (SCORAD > 30). After determination of the IPD, patients randomized into one of the three treatment arms. The patients received 15 treatments (5 times per week) without any additional therapy except for topical skin care. RESULTS: After 15 treatments the "high dose" and "medium dose" groups showed a statistically significant reduction of the SCORAD. No significant reduction of the SCORAD was observed in the "low dose" group. All three treatment arms displayed no statistically significant changes in the IgE and ECP levels and in the number of eosinophils in the peripheral blood. The UVA1 therapy was well tolerated by all patients. No side effects were observed. CONCLUSIONS: This study suggests that both the "high dose" and the "medium dose" regimens are effective in the treatment of patients with acutely exacerbated atopic dermatitis. PMID- 11405162 TI - [Intensive patient education and treatment program for young adults with atopic eczema]. AB - BACKGROUND AND OBJECTIVE: By means of a 2-week intensive multidisciplinary training & treatment course in small groups (ISBP), young adults with atopic dermatitis may be able to achieve better self-management of their disease and reduce their number of doctor visits. METHODS: Patients aged 18-35 with moderate to severe atopic dermatitis (SCORAD > 20) were randomized in a treatment (ISBP) group of n = 31 and a control group of n = 20. Follow-up was 9 months. The outcome was assessed using validated primary and secondary parameters, both specific for atopic dermatitis and more general. RESULTS: Participants in the ISBP scored significantly better at follow-up in the Marburger atopic dermatitis specific questionnaire and the self-care parameter, needed less time for medical consultations, and used more emollients without corticosteroids. Absence from work/sick leave was less at 10 weeks follow-up, but equal at 9 months. CONCLUSIONS: The ISBP program can be judged successful because both the patients and their doctors perceive their interactions as more efficient, less time time consuming and more satisfying. PMID- 11405163 TI - [Hereditary C1 esterase inhibitor deficiency type I. Divergence of clinical symptoms and laboratory chemical findings]. AB - A 35- and a 29-year-old woman presented with longstanding recurrent angioedema refractory to therapy with steroids and antihistaminic drugs. Laboratory data revealed in both cases a functional and immunohistochemical deficiency of C1 esterase-inhibitor protein (C1-INH). Further investigations of their families showed some individuals with the same pathological findings without any clinical symptoms. Thus we identified striking differences in degree of severity of clinical features in hereditary C1-INH-deficiency type I. These case-reports emphasize the importance of knowing further diagnostic steps in angioedema refractory to therapy with steroids and antihistamines drugs. Furthermore we give a picture of the different clinical courses of C1-INH-deficiency type I and underline that there is no correlation between clinical symptoms and laboratory findings. PMID- 11405164 TI - [Circumscribed myxedema of the upper and lower extremities--a rate cutaneous manifestation of Basedow disease. Case report and review of the literature]. AB - A 63 year old man presented with circumscribed myxedema surprisingly involving both the arms and legs. He had a history of thyroiditis with hyperthyroidism and an infiltrative ophthalmopathy leading to the diagnosis of Graves disease. At the time of presentation the patient had already been treated with radioactive iodine and undergone endonasal orbital decompression. We review the pathogenesis and treatment of Graves disease and discuss the first therapeutic attempt with PUVA bath therapy. PMID- 11405165 TI - [Drug reaction to terbinafine simulating an acute generalized exanthematous pustulosis]. AB - Oral terbinafine (Lamisil) is a widely used and effective therapy of dermatophyte infections. A 20-year-old patient was treated with oral terbinafine for an inguinal-scrotal fungal infection and developed an acute generalized exanthematic pustulosis (AGEP) and fever. Histological examination of skin biopsy revealed subcorneal and spongiform pustules. The diagnosis of an AGEP induced by terbinafine was established based on the history, clinical manifestations, and laboratory findings. Clinical features, differential diagnosis, causative drugs, and the therapy of AGEP are discussed and the literature on cases of terbinafine induced AGEP is reviewed. PMID- 11405166 TI - [Substitution of dehydroepiandrosterone (DHEA) in male climacteric?]]. PMID- 11405167 TI - [The graves of well-known dermatologists]. AB - The location and condition of the graves of 48 famous dermatologists are identified. Brief personal data is presented for all the physicians. In seven instances, more detailed information about the dermatologists and their tombstones is provided to illustrate the value of graves as culture sites worthy of both honor and preservation. PMID- 11405168 TI - [Addendum to the contribution by Rupec, RA, et al.: Erythema palmare hereditarium]. PMID- 11405169 TI - [Periorbital hyperpigmentation. Bilateral nevus of Ota]. PMID- 11405170 TI - [Therapy of varicella zoster and herpes simplex virus-induced diseases. 1: Virustatic agents]. PMID- 11405171 TI - [Why do malignant tumors metastasize in an organ-specific manner?]. PMID- 11405172 TI - Gemcitabine in the second-line therapy of patients with carcinoma of unknown primary site: a phase II trial of the Minnie Pearl Cancer Research Network. AB - The purpose of this study was to evaluate the activity of single-agent gemcitabine in previously treated patients with carcinoma of unknown primary site. Between January 1997 and October 1998, 39 patients were enrolled in this multicenter Phase II trial performed in the Minnie Pearl Cancer Research Network. Twenty-seven patients (69%) had adenocarcinoma or poorly differentiated adenocarcinoma; 35 patients (90%) had previously received treatment with chemotherapy regimens containing both a platinum agent and a taxane. Only 21% of patients had ever responded to previous therapy. Gemcitabine 1000 mg/m2 was administered intravenously on days 1, 8, and 15 of each 28-day course. Three of 36 evaluable patients (8%) had partial responses, and 9 patients (25%) had minor responses or stable disease with improved symptoms. The median time to progression for patients with partial responses or stable disease/improved symptoms was 5 months. Treatment was well tolerated, with uncommon grade 3 or 4 toxicity. Gemcitabine produced a low objective response rate in this refractory patient population, although approximately one-third of patients experienced symptomatic improvement. Treatment with gemcitabine was well tolerated. Because gemcitabine has activity against a variety of adenocarcinomas, further evaluation of this agent as part of first-line therapy for patients with carcinoma of unknown primary site is appropriate. PMID- 11405173 TI - Adjuvant postoperative 5-fluorouracil chemotherapy combined with pelvic radiation for rectal cancer: results from an Asian population. AB - Prospective randomized clinical trials have shown the effectiveness of combined adjuvant 5-fluorouracil-based chemotherapy and radiotherapy after surgical resection of rectal cancer. To assess toxicity of this therapy, prospective data were collected from 236 Asian rectal cancer patients treated with combined 5 fluorouracil-based chemotherapy and radiotherapy after surgery. Almost 82% of patients completed planned therapy. Grade 3 and 4 diarrhea, stomatitis, and granulocytopenia occurred in approximately 18-21% of patients. There were two treatment-related deaths from granulocytopenia and sepsis. With median follow-up of 3.5 years, median disease-free and overall survival was 75 and 88 months, respectively. In conclusion, combined adjuvant 5-fluorouracil-based chemotherapy and radiotherapy after surgical resection of rectal cancer is tolerable in Asian patients with moderate toxicity. PMID- 11405174 TI - Quantitation of interferon regulatory factor transcripts in patients with acute myeloid leukemia. AB - Interferon regulatory factors IRF-1 and IRF-2, the two mutually antagonistic factors, fluctuate during the cell cycle and play an important role in normal and neoplastic growth processes. The relative levels of these two transcripts were analyzed in 5 normal and 43 acute myeloid leukemia (AML) bone marrow (BM) specimens by a semiquantitative RT-PCR method. IRF-1 and IRF-2 cDNA sequences were coamplified using primers that were designed to span regions of high homology between the genes. Each primer can anneal equally to both IRF-1 and IRF 2 sequences. Hence, the relative amount of amplified products from each cDNA species provides an estimation of proportional concentration of the RNA transcripts in the test sample. Results indicate expression of both the transcripts on all the leukemia and lymphoma cell lines tested, normal and AML BM. Significantly higher IRF-1:IRF-2 ratio was observed in normal as compared to AML BM (p = 0.007). There was no correlation with clinical factors such as FAB subtype. A single dose of amifostine or three daily doses of recombinant IL-4 were administered to 5 and 8 AML patients, respectively. The changes in the expression of these transcripts were studied prior to administration of the agent (d0) and after 3 days (d3). IL-4 treatment showed significant increase in the IRF 1:IRF-2 ratio in 4 of 8 patients (p = 0.05); amifostine treatment did not show any appreciable change. PMID- 11405175 TI - Is there a potential role for serum cardiac troponin I as a marker for myocardial dysfunction in pediatric patients receiving anthracycline-based therapy? A pilot study. AB - Serum cardiac troponin I (cTnI) levels have been reported to have high specificity and sensitivity to acute myocardial infarction and coronary ischemic syndromes in adult patients. Our goal was to evaluate the usefulness of serum cTnI in the early diagnosis of cardiac injury from anthracyclines, and to compare these values with echocardiographic findings of cardiac dysfunction. In this prospective study, children being treated on several Children's Cancer Group protocols underwent measurement of shortening fraction (SF), ejection fraction (EF), and serum cTnI levels prior to anthracycline therapy. Sequential serum cTnI levels were then measured along with regularly scheduled echocardiograms with progressively increasing doses of anthracyclines. Fifteen children with median age of 5.75 years (range, 15 months to 15.5 years) at diagnosis were evaluated. Anthracycline doses ranged from 11.72 mg/kg (in patients < 3 years of age) to 375 mg/m2. All but one patient had normal cTnI levels. His level measured at 1.7 ng/ml after 315 mg/m2, but was normal on follow-up testing. Initial SF ranged from 32 to 48%, and EF from 60 to 80%. On follow-up, SF and EF ranged from 30 to 41% and 55 to 70%, respectively. Both SF and EF were significantly lower (p < 0.001) as compared to the initial values. Despite this, all patients remained clinically asymptomatic from the cardiac standpoint. We did not observe elevations of serum cTnI levels in clinically asymptomatic children who received anthracycline therapy up to doses of 375 mg/m2. Does this mean that cardiac injury has not occurred? The possibility of assay sensitivity and the timing of serum sampling and echocardiograms may be important. In addition, larger sample size or longer follow-up may be helpful to determine if higher doses or symptomatic patients potentially have elevations in cTnI levels. PMID- 11405176 TI - Mitomycin C and cisplatin enhanced the antitumor activity of p53-expressing adenovirus in cervical cancer cells. AB - This study investigated the enhanced antitumor activity of Ad5-p53 in combination with mitomycin C (MMC) or cisplatin (DDP) in cervical cancer cell lines SiHa and C-33A. MMC and DDP inhibited the growth of SiHa and C-33A cells in a dose dependent manner, and the combination of MMC or DDP with Ad5-p53 showed a stronger growth inhibition than those treated with either Ad5-p53, MMC, or DDP alone. As evidenced by the formation of the approximately 200 bp DNA ladder and the appearance of sub-G1 peak, both MMC and DDP induced apoptosis in cervical cancer cells. Western blot analysis of p53 showed that MMC/DDP did not induce the increase of p53 protein in SiHa cells nor the increase of the cellular and nuclear p53 protein in Ad5-p53 transfected Saos-2 cells. Taken together, these results suggested that the combination of Ad5-p53 and MMC/DDP may serve as an effective therapeutic regime for human cervical cancer treatment. PMID- 11405177 TI - Peripheral neuropathy in cancer patients. PMID- 11405178 TI - Simian T-cell lymphotropic Viruses (STLVs) and lymphomas in African nonhuman primates. PMID- 11405179 TI - BRCA1 and prostate cancer. AB - The breast cancer susceptibility gene BRCA1 on chromosome 17q21 encodes an 1863 amino acid protein that is important for normal embryonic development. Germline mutations of this gene are linked to a significantly increased lifetime risk for breast and/or ovarian cancer, and recent studies suggest that the same may be true for prostate cancer. Several activities that may contribute to the tumor suppressor function of BRCA1 have been identified via in vitro and experimental animal studies. These include (i) regulation of cell proliferation; (ii) participation in DNA repair/recombination processes related to the maintenance of genomic integrity; (iii) induction of apoptosis in damaged cells; and (iv) regulation of transcription. A second breast cancer susceptibility gene (BRCA2) operates in some of the same molecular pathways as BRCA1, and mutations of this gene predispose to breast and ovarian cancer and probably to other tumor types, including prostate cancer. Finally, recent studies from our laboratory suggest that BRCA1 modulates proliferation, chemosensitivity, repair of DNA strand breaks, apoptosis induction, and expression of certain key cellular regulatory proteins (including BRCA2 and p300) in human prostate cancer cells. These activities are consistent with a putative prostate tumor suppressor function of BRCA1. PMID- 11405180 TI - Contemporary drug therapy in palliative care: new directions. PMID- 11405181 TI - Pegylated liposomal doxorubicin: metamorphosis of an old drug into a new form of chemotherapy. AB - Pegylated liposomal doxorubicin (Doxil, Caelyx) is a formulation of doxorubicin in poly(ethylene glycol)-coated (stealth) liposomes with a prolonged circulation time and unique toxicity profile. We review the preclinical and clinical pharmacology as well as recent clinical data obtained in specific cancer types. Doxil liposomes retain the drug payload during circulation and accumulate preferentially in tissues with increased microvascular permeability, as often is the case of tumors. Doxil toxicity profile is drastically different from that of doxorubicin, and is characterized by dominant and dose-limiting mucocutaneous toxicities, mild myelosuppression, minimal alopecia, and no apparent cardiac toxicity. Although the single maximum tolerated dose (MTD) of Doxil is actually lower than that of conventionally administered doxorubicin, the cumulative MTD dose of Doxil may be substantially greater than that of free doxorubicin. Doxil is probably one of the most active agents in AIDS-related Kaposi's sarcoma and has a definite role in management of recurrent ovarian cancer. The potential of Doxil in the treatment of other cancer types and the opportunities it offers in combination with other drugs and therapeutic modalities are under active investigation. PMID- 11405182 TI - Literacy, comprehension, and informed consent in clinical research. AB - Institutional review boards (IRBs) and informed consent have recently come under increasing scrutiny. The provision of appropriate and understandable information and its comprehension are key elements of the informed consent process. This article examines how literacy and other factors might impact on comprehension of the information provided. Better understanding of these issues and how to best address them are crucial to allowing clinical research subjects to participate as well-informed and willing partners in scientific inquiry. PMID- 11405183 TI - Treatment of tuberculous peritonitis. PMID- 11405184 TI - Empiric initial therapy and dose of vancomycin. PMID- 11405185 TI - Using the electroretinogram to detect and monitor the retinal toxicity of anticonvulsants. PMID- 11405186 TI - [Association between breast and colorectal cancer and polyps. Screening and epidemiological study]. AB - BACKGROUND: Studies regarding the associations between different types of cancer in the same patient are very few and not always come to the same conclusions. Several hypothesis are suggested and particularly genetic and socioeconomical ones seem to offer an interpretation of this issue. Early detection of a second neoplasm allows to improve prognosis and survival. The knowledge of correlations between tumors help to select a population, with a high risk to develop a second cancer, to be included in a screening program. Nowadays thanks to early detection of breast cancer, ten years survival is more than 75%. Women who had breast cancer now live longer and so could have a higher risk to develop a second cancer. METHODS: From September 1998 to September 1999 in our Department 71 patients operated for breast cancer, underwent screening colonscopy. No patients refused to be included in the study. Mean age was 61 years (range 36-87). Each patient had a clinician interview in order to explain the goals of the study. RESULTS: Results show that among all patients 3 (4.2%) presented a history of colon cancer, 18.3% (13 cases) presented large bowel polyps. In 84.60% patients (11 cases) polyps were found not over 40 cm. This study shows that 93% of patients (66 cases) had a relative with cancer history. CONCLUSIONS: Our results compared with those of other authors seem to show an increased risk for breast cancer patients in developing polyps or colon cancer, so we suggest to insert sigmoidoscopy in standard follow-up of breast cancer patients. PMID- 11405187 TI - [Ambulatory surgical treatment for breast carcinoma]. AB - BACKGROUND: The aim of the study is to demonstrate the feasibility and the oncologic effectiveness of quadrantectomy plus sentinel node biopsy performed under local anesthesia, and to demonstrate the economic and psychologic advantages. METHODS: From October 1996 to March 2000, 71 patients affected with clinical T1 N0 breast cancer, underwent quadrantectomy or tumor resection plus sentinel node biopsy and clinically suspicion axillary nodes biopsy, under local anesthesia at the Casa di Cura "Villa Mafalda" in Rome. RESULTS: Twenty tumors were T1a, 26 T1b e 25 T1c. A mean of 2 sentinel nodes (range 1-4) and a mean of 8 axillary nodes were removed during the procedure. In 2 cases sentinel nodes were not identified. Intraoperative histologic examination showed metastatic sentinel nodes in 11 cases. An axillary node dissection was performed in all cases (>12 nodes) and no other metastatic nodes were found. In all patients clinically suspected nodes were removed. In two cases no evidence of metastasis was found in sentinel nodes, while histologic examination revealed in a patient micrometastasis in one node, and in another patient two metastatic nodes. CONCLUSIONS: Fifty-three patients rated the overall surgical, anesthetic and recovery experience as "very satisfactory", 13 "satisfactory" and 5 "unsatisfactory". Patients typically expressed their pleasure at the possibility to return home and stressed the ease of recovery. PMID- 11405188 TI - Gene monitoring of surface-activated monocytes in circulating whole blood using duplex RT-PCR. AB - Previous studies have shown that blood-contacting materials activate blood monocytes. The aim of this study is to evaluate a highly sensitive technique for detecting changes at the mRNA level in circulating monocytes and to find suitable "gene markers" for assessing the hemocompatibility of biomaterials. Human blood was recirculated in a modified Chandler Loop formed of test tubes. Five groups of biomaterials were compared. Monocytes were separated by using magnetic beads, and gene expression analyses were performed using RT-PCR. We investigated the mRNA expression of stress proteins (hsp70) and inflammatory markers (IL-1alpha, IL 1beta, IL-6, TNF-alpha). mRNAs for cytokines were highly upregulated in LPS stimulated monocytes. Hsp70 transcripts were upregulated after heat shock but were not influenced by treatment with LPS. The gene expression of the cytokines was affected by various biomaterials. The intensity of the inflammatory response increased in the following order: heparin coatings (PVC) < uncoated PVC < silicone, polypeptide-coated PVC. No change was seen in the expression of the hsp70 genes. An inhibitory effect of systemic heparin on surface-activated monocytes was observed for the mRNA expression of the cytokines. The recirculation of human blood in an in vitro model in combination with the immunomagnetic separation of monocytes and the Duplex RT-PCR method is a powerful tool for getting reliable results. Our data demonstrate that hsp70 genes cannot be recommended for short-term, biomaterial-induced mRNA expression studies. Conversely, mRNAs for IL-1alpha, IL-1beta, IL-6, and TNF-alpha were expressed in a material-dependent manner. Thus our model provides an effective tool for assessing the hemocompatibility of biomaterials before their clinical application and it also can serve as a safety control during the industrial manufacturing process. This method can be applied to various blood cell populations and numerous gene expression studies and may enable a more fundamental understanding of the biologic processes involved in blood-material interactions. PMID- 11405189 TI - Physiological load imposed on elite soccer referees during actual match play. AB - BACKGROUND: The aim of this study as to describe the work-rate profile and cardiovascular stress of Italian high level soccer referees (n = 18). METHODS. EXPERIMENTAL DESIGN: The referees were examined during official games of the 1992 1993 first division Italian championship (serie A). PARTICIPANTS: subjects were all experienced top level referees enrolled in the CAN (Commissione Arbitri Nazionali) and thus officiating in the serie A e B Italian championships. Mean age of the referees considered here was 37.5 +/- 2.14 years. MEASURES: heart rates were monitored by short-range radio telemetry. Match analysis was performed with methodologies similar to that reported by Ohashi et al. (1988). RESULTS: Average match distance was 11376 +/- 1600 m (7818-13063) and no overall differences were observed between halves (p > 0.05). Nevertheless less distances were covered running backward and side-wards during the second half compared to the first half (p < 0.05). Referees, on average, covered 41.7% of the whole match distance at speeds faster than 13.1 km x h(-1). Heart rates attained 89.1% of the estimated maximal over the duration of a full game and no difference was observed between halves (p = 0.72). Mean heart rates of the first 15 min of play were lower than the other two sections of the first half (p < 0.05). CONCLUSIONS: Match analysis revealed the intermittent nature of the referees' activities. Their intensity varied from situation to situation, frequently reaching near maximal intensity. However, sprint bouts never lasted for more than few seconds (2-4 sec). Top level soccer refereeing places high physiological demands on the official thus specific training and fitness assessment are needed. PMID- 11405190 TI - Guns killing our children: a status report. PMID- 11405191 TI - Raising kids, grandparents bear a burden. PMID- 11405192 TI - A person of one's time. PMID- 11405193 TI - Is it Kawasaki disease? PMID- 11405194 TI - Statins and bone formation. AB - The main therapy needed most in the bone field is an anabolic agent for the treatment of osteoporosis. Current drugs on the market, which included bisphosphonates, calcitonin, estrogen and related compounds, vitamin D analogues trabecular microarchitecture. Therefore, it would be desirable to have a satisfactory and universally and iprifalvone, are essentially bone resorption inhibitors that mainly act to stabilize bone mass. Patients with established osteoporosis have lost more than 50% of their bone mass at critical sites in the skeleton, and more over have marked disruption of acceptable drug that would stimulate new bone formation and correct this disturbance of trabecular microarchitecture characteristic of established osteoporosis. Recently inhibitors of the enzyme 3-hydroxy-3-methylglutaryl coenzyme A (HMG CoA) reductase, which controls the first step in the biosynthesis of cholesterol, have been shown to stimulate bone formation in rodents both in vitro and in vivo. The effect is associated with an increased expression of the bone morphogenetic protein-2 (BMP 2) gene in bone cells. These statins drugs are widely used agents for lowering cholesterol and reducing heart attacks, however they are also known to elicit numerous pleiotropic effects including inhibition of proliferation and migration of smooth muscle cells, inhibition of tumor growth and anti-inflammatory activity. Some of these effects have been attributed to not only to the reduction of cholesterol synthesis by inhibition of the HMG-CoA reductase enzyme but also by the concurrent reduction in downstream metabolites of the mevalonate pathway such as mevalonate, farnesyl pyrophosphate and geranylgeranyl pyrophosphate. The findings that statins are capable of increasing bone formation and bone mass in rodents suggests a potential new action for the statins, which may be beneficial in patients with established osteoporosis where marked bone loss has occurred. Recent clinical data suggests that they may reduce the risk of fracture in patients taking these drugs. However, their precise role can only be determined by appropriate randomized clinical trials, which demonstrate their efficacy in this regard in patients. PMID- 11405195 TI - [Images in medicine. Thromboembolic renal infarct]. PMID- 11405196 TI - [Images in medicine. An abnormal preoperative radiography of the thorax]. PMID- 11405197 TI - Evidence-based medicine (EBM) in the treatment of hypertension in older patients. PMID- 11405198 TI - Orientation clinics. PMID- 11405199 TI - Staining the anterior capsule. PMID- 11405200 TI - Combined technique: AK and LASIK. PMID- 11405201 TI - Misleading tonometry after LASIK. PMID- 11405202 TI - Chronic or saccular endophthalmitis: diagnosis and management. PMID- 11405203 TI - Postoperative myopia with subsequent hyperopic shift after phacoemulsification and multifocal IOL implantation. PMID- 11405204 TI - Women's request for Cesarean. PMID- 11405205 TI - Advance in contrast agents, reporters, and detection. PMID- 11405206 TI - Screening for congenital hypothyroidism (CH) among Filipino newborn infants. Philippine Newborn Screening Study Group. AB - From June 1996 to June 1998 a total of 62.841 newborn infants were screened for congenital hypothyroidism with thyroid stimulating hormone assay as a primary test. The method used was an immunofluorescent assay using the DELFIA TSH Kit on dried blood specimens collected by heelprick on filter paper. All infants with TSH values greater than 20 microU/ml were retested. If the results remained abnormally high, confirmatory testing was done by radioimmunoassay. All infants who were confirmed to be hypothyroid were referred to pediatric endocrinologists for initial management. The overall weighted incidence of congenital hypothyroidism obtained in this study was 0.000277 (95% CI; 0.000122 - 0.000432) or 1:3,610 which may be higher than that reported by most screening programs worldwide. The recall rate was 0.16%. The higher recall rate may be explained by early testing in a number of cases and by the possibility of iodine deficiency in some of the mothers. On the basis of the results of this study, we would recommend (1) screening on a greater number of infants to verify the incidence of CH and (2) establishing normal TSH values at different hours of life to improve our recall rate. PMID- 11405207 TI - Screening for galactosemia: Philippines experience. Newborn Screening Study Group. AB - Galactosemia is an inborn error of galactose metabolism due to a deficiency of any of the galactokinase, galactose-1-phosphate uridyl transferase (GALT), or epimerase enzymes. The Philippines, with its pilot newborn screening project, has been screening for this disorder for 2 years now. A total of 62,841 babies have been screened using the galactose and galactose-1-phosphate spot test. Confirmatory testing is done by the newborn screening laboratory of the The New Children's Hospital in Westmead, Australia. Two cases of galactosemia: 1 classical galactosemia and 1 galactokinase deficiency have so far been confirmed. Clinical review, problems encountered, and management are described. Long-term outcome of these patients, however, is yet to be determined. PMID- 11405208 TI - Web alert. PMID- 11405209 TI - Global tropospheric ozone dynamics. Part II: Numerical modelling of tropospheric ozone variability. AB - Various causes of tropospheric changes have been considered in Part I in connection with the analysis of observation data. It is clear however, that the principal instrument for understanding numerous and often interacting causes of ozone changes is numerical modelling. A review of the current status of the numerical modelling has been made for the variability of the ozone concentration in the troposphere. Observation data on tropospheric ozone and relevant numerical modelling results show that a necessity exists to get more adequate global observational data. PMID- 11405210 TI - [Improving risk reporting in public health: no time for delays]. PMID- 11405211 TI - Granulocyte colony-stimulating factor-induced sickle cell crisis and multiorgan dysfunction in a patient with compound heterozygous sickle cell/beta+ thalassemia. PMID- 11405212 TI - Alloimmunization in Hong Kong southern Chinese transfusion-dependent thalassemia patients. PMID- 11405213 TI - X chromosome inactivation ratios in female carriers of X-linked sideroblastic anemia. PMID- 11405214 TI - Prognostic value of p16(INK4a) gene deletions in pediatric acute lymphoblastic leukemia. PMID- 11405215 TI - Vaccine development: from empirical medicine to molecularly designed therapy. PMID- 11405216 TI - Dendritic cells for specific cancer immunotherapy. AB - The characterization of tumor-associated antigens recognized by human T lymphocytes in a major histocompatibility complex (MHC)-restricted fashion has opened new possibilities for immunotherapeutic approaches to the treatment of human cancers. Dendritic cells (DC) are professional antigen presenting cells that are well suited to activate T cells toward various antigens, such as tumor associated antigens, due to their potent costimulatory activity. The availability of large numbers of DC, generated either from hematopoietic progenitor cells or monocytes in vitro or isolated from peripheral blood, has profoundly changed pre clinical research as well as the clinical evaluation of these cells. Accordingly, appropriately pulsed or transfected DC may be used for vaccination in the field of infectious diseases or tumor immunotherapy to induce antigen-specific T cell responses. These observations led to pilot clinical trials of DC vaccination for patients with cancer in order to investigate the feasibility, safety, as well as the immunologic and clinical effects of this approach. Initial clinical studies of human DC vaccines are generating encouraging preliminary results demonstrating induction of tumor-specific immune responses and tumor regression. Nevertheless, much work is still needed to address several variables that are critical for optimizing this approach and to determine the role of DC-based vaccines in tumor immunotherapy. PMID- 11405217 TI - Intracellular bacteria as targets and carriers for vaccination. AB - In this review we discuss intracellular bacteria as targets and carriers for vaccines. For clarity and ease of comprehension, we focus on three microbes, Mycobacterium tuberculosis, Listeria monocytogenes and Salmonella, with an emphasis on tuberculosis, one of the leading causes of death from infectious disease. Novel vaccination strategies against these pathogens are currently being considered. One approach favors the use of live attenuated vaccines and vaccine carrier strains thereof, either for heterologous antigen presentation or DNA vaccine delivery. This strategy includes both the improvement of attenuated vaccine strains as well as the 'de novo' generation of attenuated variants of virulent pathogens. An alternative strategy relies on the application of subunit immunizations, either as nucleic acid vaccines or protein antigens of the pathogen. Finally, we present a short summary of the vaccination strategies against tuberculosis. PMID- 11405218 TI - Bacteria-mediated transfer of eukaryotic expression plasmids into mammalian host cells. AB - Invasive intracellular bacteria are able to transfer eukaryotic expression plasmids into mammalian host cells in vitro and in vivo. This can be used to induce immune responses toward protein antigens encoded by the plasmid or to complement genetic defects. Plasmid transfer takes place when the recombinant bacterium dies within the host cell, either due to metabolic attenuation or induction of autolysis. Alternatively, antibiotics can be used and spontaneous transfer has also been observed, indicating that this phenomenon might also occur under physiological conditions. Plasmid transfer has been reported for Shigella flexneri, Salmonella typhimurium and S. typhi, Listeria monocytogenes and recombinant Escherichia coli, but other invasive bacteria should also share this property. In vivo attempts were mainly directed toward vaccination using shigella and salmonella as carrier. So far a wide variety of antigens have been used succesfully in mice. Often this type of immunization was superior over direct application of antigen or using the same bacterium as a heterologous carrier expressing the antigen via a prokaryotic promoter. Characterization of the host cells revealed that macrophages and dendritic cells might be responsible for immune stimulation by either expressing the antigen or cross-presenting the antigen after uptake of apoptotic antigen expressing cells. PMID- 11405219 TI - Revealing the potential of DNA-based vaccination: lessons learned from the hepatitis B virus surface antigen. AB - DNA-based vaccination is a novel technique to efficiently stimulate humoral (antibody) and cellular (T cell) immune responses to protein antigens. In DNA based vaccination, immunogenic proteins are expressed in in vivo transfected cells of the vaccine recipients in their native conformation with correct posttranslational modifications from antigen-encoding expression plasmid DNA. This ensures the integrity of antibody-defined epitopes and supports the generation of protective (neutralizing) antibody titers. Plasmid DNA vaccination is furthermore an exceptionally potent strategy to stimulate CD8+ cytotoxic T lymphocyte (CTL) responses because antigenic peptides are efficiently generated by endogenous processing of intracellular protein antigens. These key features make DNA-based immunization an attractive strategy for prophylactic and therapeutic vaccination against extra- and intracellular pathogens. In this brief review, we summarize the current state of expression vector design, DNA delivery strategies, priming immune responses to intracellular or secreted antigens by DNA vaccines and unique advantages of DNA- versus recombinant protein-based vaccines using the hepatitis B surface antigen (HBsAg) as a model antigen. PMID- 11405220 TI - Progress toward a malaria vaccine: efficient induction of protective anti-malaria immunity. AB - Malaria can be a very severe disease, particularly in young children, pregnant women (mostly in primipara), and malaria naive adults, and currently ranks among the most prevalent infections in tropical and subtropical areas throughout the world. The widespread occurrence and the increased incidence of malaria in many countries, caused by drug-resistant parasites (Plasmodium falciparum and P. vivax) and insecticide-resistant vectors (Anopheles mosquitoes), indicate the need to develop new methods of controlling this disease. Experimental vaccination with irradiated sporozoites can protect animals and humans against the disease, demonstrating the feasibility of developing an effective malaria vaccine. However, developing a universally effective, long lasting vaccine against this parasitic disease has been a difficult task, due to several problems. One difficulty stems from the complexity of the parasite's life cycle. During their life cycle, malaria parasites change their residence within the host, thus avoiding being re-exposed to the same immunological environment. These parasites also possess some distinct antigens, present at different life stages of the parasite, the so-called stage-specific antigens. While some of the stage-specific antigens can induce protective immune responses in the host, these responses are usually genetically restricted, this being another reason for delaying the development of a universally effective vaccine. The stage-specific antigens must be used as immunogens and introduced into the host by using a delivery system that should efficiently induce protective responses against the respective stages. Here we review several research approaches aimed at inducing protective anti-malaria immunity, overcoming the difficulties described above. PMID- 11405221 TI - Peptide vaccines and peptide libraries. AB - Synthetic immunogens, containing built-in adjuvanticity, B cell, T helper cell and CTL epitopes or mimotopes, are ideal and invaluable tools to study the immune response with respect to antigen processing and presentation. This serves as a basis for the development of complete and minimal vaccines which do not need large carrier proteins, further adjuvants, liposome formulations or other delivery systems. Combinatorial peptide libraries, either completely random or characterized by one or several defined positions, are useful tools for the identification of the critical features of B cell epitopes and of MHC class I and class II binding natural and synthetic epitopes. The complete activity pattern of an O/Xn library with hundreds of peptide collections, each made up from billions of different peptides, represents the ranking of amino acid residues mediating contact to the target proteins of the immune system. Combinatorial libraries support the design of peptides applicable in vaccination against infectious agents as well as therapeutic tumour vaccines. Using the principle of lipopeptide vaccines, strong humoral and cellular immune responses could be elicited. The lipopeptide vaccines are heat-stable, non-toxic, fully biodegradable and can be prepared on the basis of minimized epitopes by modern methods of multiple peptide synthesis. The lipopeptides activate the antigen-presenting macrophages and B cells and have been recently shown to stimulate innate immunity by specific interaction with receptors of the Toll family. PMID- 11405222 TI - Defined synthetic vaccines. AB - Although vaccines have proven very successful in preventing certain infectious diseases, progress in the field has been slowed by the tediousness of developing classical vaccines consisting of whole pathogens. Thus, there is great need for improvement in several areas: firstly, the range of diseases which can be treated has to be expanded. Secondly, antigens have to be defined to make the use of whole pathogens as antigen obsolete. And thirdly, new adjuvants have to be developed which show low toxicity, high potency and are also able to drive the immune response in the desired direction. Ideally, a vaccine would only consist of well-characterized, synthetic materials. This review summarizes the different approaches for the development of completely defined synthetic vaccines. PMID- 11405223 TI - Antimicrobial peptides: properties and applicability. AB - All organisms need protection against microorganisms, e. g. bacteria, viruses and fungi. For many years, attention has been focused on adaptive immunity as the main antimicrobial defense system. However, the adaptive immune system, with its network of humoral and cellular responses is only found in higher animals, while innate immunity is encountered in all living creatures. The turning point in the appreciation of the innate immunity was the discovery of antimicrobial peptides in the early eighties. In general these peptides act by disrupting the structural integrity of the microbial membranes. It has become clear that membrane-active peptides and proteins play a crucial role in both the innate and the adaptive immune system as antimicrobial agents. This review is focused on the functional and structural features of the naturally occurring antimicrobial peptides, and discusses their potential as therapeutics. PMID- 11405224 TI - G-quadruplex DNA structures--variations on a theme. AB - To be functional, nucleic acids need to adopt particular three-dimensional structures. For a long time DNA was regarded as a rigid and passive molecule with the sole purpose to store genetic information, but experimental data has now accumulated that indicates the full dynamic repertoire of this macromolecule. During the last decade, four-stranded DNA structures known as G-quadruplexes, or DNA tetraplexes, have emerged as a three-dimensional structure of special interest. Motifs for the formation of G-quadruplex DNA structures are widely dispersed in eukaryotic genomes, and are abundant in regions of biological significance, for example, at telomeres, in the promoters of many important genes, and at recombination hotspots, to name but a few in man. Here I explore the plethora of G-quadruplex DNA structures, and discuss their possible biological functions as well as the proteins that interact with them. PMID- 11405225 TI - The role of heat shock proteins and their receptors in the activation of the immune system. AB - Heat shock proteins (HSPs) have been described as potent tumor vaccines in animal models and are currently studied in clinical trials. The underlying immune response relies on immunogenic peptides that the HSPs have acquired intracellularly by interfering with the classical antigen processing pathways. There have been numerous reports shedding light on how HSPs are able to gain this function and a number of important requirements for HSP-mediated specific immunity have been described: first, the ability of HSPs to bind immunogenic peptides. Second, the acquisition of HSPs by specialized antigen presenting cells with efficient antigen processing pathways capable of inducing cellular immune responses. Third, the existence of specific receptors on the surfaces of antigen presenting cells, allowing efficient and rapid uptake of HSP-peptide complexes from the extracellular fluid. And fourth, the ability of heat shock proteins to activate antigen presenting cells, enabling the latter to prime cytotoxic T cell responses against the peptides associated to HSPs. PMID- 11405226 TI - Transcriptional repression mediated by the KRAB domain of the human C2H2 zinc finger protein Kox1/ZNF10 does not require histone deacetylation. AB - The KRAB domain of human Kox1, a member of the KRAB C2H2 zinc finger family, confers strong transcriptional repressor activities even to remote promoter positions. Here, HDAC inhibitors were used to demonstrate that histone deacetylation is not required for mediating transcriptional repression of KRAB zinc finger proteins. Two reporter systems with either stably integrated or transiently transfected templates, both under control of strong viral promoters, were analyzed. Under all circumstances, HDAC inhibition did not alter the repression potential of the KRAB domain. In case of the stably integrated luciferase reporter gene system, neither expression levels of the KRAB fusion protein nor complex formation with its putative co-repressor TIF1beta were significantly changed. Furthermore, the TIF1beta/KRAB complex was devoid of mSin3A and HDAC1. In the transient transfection system, the transcriptional repression induced by TIF1beta and HP1alpha was not diminished by HDAC inhibitors, whereas the repressory activity of TIF1alpha was significantly affected. Thus, KRAB, TIF1beta and HP1alpha are likely to be functionally linked. In conclusion, HDAC activity is not essential for the strong transcriptional repressor activity mediated by the KRAB domain of Kox1 in particular and, presumably, by KRAB domains in general. This feature might be helpful in identifying and characterizing target genes under the control of PMID- 11405227 TI - Structure and evolution of 4-coumarate:coenzyme A ligase (4CL) gene families. AB - The phenylpropanoid enzyme 4-coumarate:coenzyme A ligase (4CL) plays a key role in general phenylpropanoid metabolism. 4CL is related to a larger class of prokaryotic and eukaryotic adenylate-forming enzymes and shares several conserved peptide motifs with these enzymes. In order to better characterize the nature of 4CL gene families in poplar, parsley, and tobacco, we used degenerate primers to amplify 4CL sequences from these species. In each species additional, divergent 4CL genes were found. Complete cDNA clones for the two new poplar 4CL genes were obtained, allowing examination of their expression patterns and determination of the substrate utilization profile of a xylem-specific isoform. Phylogenetic analysis of these genes and gene fragments confirmed previous results showing that 4CL proteins fall into two evolutionarily ancient subgroups . A comparative phylogenetic analysis of enzymes in the adenylate-forming superfamily showed that 4CLs, luciferases, and acetate CoA ligases each form distinct clades within the superfamily. According to this analysis, four Arabidopsis 4CL-like genes identified from the Arabidopsis Genome Project are only distantly related to bona fide 4CLs or are more closely related to fatty acid CoA ligases, suggesting that the three Arabidopsis 4CL genes previously characterized represent the extent of the 4CL gene family in this species. PMID- 11405228 TI - Inhibition of hepatitis B virus by hammerhead ribozyme targeted to the poly(A) signal sequence in cultured cells. AB - The deviant poly(A) signal of hepatitis B virus (HBV) not only controls the formation of the 3' end of all the viral RNA, but is also crucial for HBV replication. Hence, a cis-releasing hammerhead ribozyme (RzA) targeted to the poly(A) signal region of HBV subtype adr was investigated for its antiviral effects. In vitro, RzA cleaved HBV RNA at its target site up to 70%, while the disabled ribozyme (dRzA), which had a one-base mutation in the catalytic site, did not cleave the target RNA at all. When the ribozymes were cotransfected into HepG2 cells with the HBV genome-containing plasmid p3.6II, the wild-type ribozyme RzA could effectively decrease HBV RNA levels and inhibit HBV replication, whereas its disabled form, dRzA, had much weaker effects, indicating that the active catalytic domain of the hammerhead ribozyme could markedly increase the extent of antisense-mediated inhibition. In addition, there was a gradient of effectiveness: the higher the amount of released ribozyme, the more the reduction in target HBV RNA in cells as well as progeny DNA reduction. These results suggest the possibility of the hammerhead ribozyme RzA to be used for the gene therapy of HBV infection. PMID- 11405229 TI - Chemical accessibility of 18S rRNA in native ribosomal complexes: interaction sites of mRNA, tRNA and translation factors. AB - During protein synthesis the ribosome interacts with ligands such as mRNA, tRNA and translation factors. We have studied the effect of ribosome-ligand interaction on the accessibility of 18S rRNA for single strand-specific modification in ribosomal complexes that have been assembled in vivo, i. e. native polysomes. A comparison of the modification patterns derived from programmed and non-programmed ribosomes showed that bases in the 630- and 1060 loops (530- and 790-loops in E. coli) together with two nucleotides in helices 33 and 34 were protected from chemical modification. The majority of the protected sites were homologous to sites previously suggested to be involved in mRNA and/or tRNA binding in prokaryotes and eukaryotes, implying that the interaction sites for these ligands are similar, if not identical, in naturally occurring programmed ribosomes and in in vitro assembled ribosomal complexes. Additional differences between programmed and non-programmed ribosomes were found in hairpin 8. The bases in helix 8 showed increased exposure to chemical modification in the programmed ribosomes. In addition, structural differences in helices 36 and 37 were observed between native 80S run-off ribosomes and 80S ribosomes assembled from isolated 40S and 60S subunits. PMID- 11405230 TI - C-terminal peptides of interleukin-6 modulate the expression of junB protooncogene and the production of fibrinogen by HepG2 cells. AB - Interleukin-6 (IL-6) is a 185 amino acid residue helical cytokine with various biological activities (e. g. B cell development, acute phase reaction). We have investigated the role of the 168-185 C-terminal region of IL-6 in the induction of fibrinogen synthesis and expression of junB mRNA using synthetic peptides corresponding to this region. Circular dichroism spectroscopy data suggest that even truncated peptides have a strong tendency to adopt an ordered conformation. Peptides were tested alone or in combination with recombinant hIL-6 on an IL-6 responsive human hepatoma HepG2 cell line. The expression of the protooncogene junB monitored by competitive RT-PCR represents an early, while the fibrinogen production detected by sandwich ELISA a late, marker of IL-6 initiated events. We found that peptides--depending on their structure--modulate spontaneous as well as IL-6 induced fibrinogen production and/or mRNA expression of junB by exhibiting inhibition (in the presence of IL-6) or stimulation (in the absence of IL-6). PMID- 11405231 TI - Proteome analysis by three-dimensional protein separation: turnover of cytosolic proteins in hepatocytes. AB - We performed a three-dimensional separation of pulse-chase dual-labelled rat liver cytosolic proteins using hydrophobic interaction chromatography, isoelectric focusing, and SDS gel electrophoresis. Due to very different expression rates but similar size and pI of rat liver cytosolic proteins, we demonstrate the impossibility of successful two-dimensional separations of such complex protein mixtures. A pre-fractionation of proteins by hydrophobic interaction chromatography is therefore recommended prior to two-dimensional gel electrophoresis. Our studies confirmed the correlation between protein turnover rates and surface hydrophobicity. PMID- 11405232 TI - Structural intermediates in the putative pathway from the cellular prion protein to the pathogenic form. AB - The conversion of the alpha-helical, protease sensitive and noninfectious form of the prion protein (PrP(C)) into an insoluble, protease resistant, predominantly beta-sheeted and infectious form (PrP(Sc)) is the fundamental event in prion formation. In the present work, two soluble and stable intermediate structural states are newly identified for recombinant Syrian hamster PrP(90-231) (recPrP), a dimeric alpha-helical state and a tetra- or oligomeric, beta-sheet rich state. In 0.2% SDS at room temperature, recPrP is soluble and exhibits alpha-helical and random coil secondary structure as determined by circular dichroism. Reduction of the SDS concentration to 0.06% leads first to a small increase in alpha-helical content, whereas further dilution to 0.02% results in the aquisition of beta sheet structure. The reversible transition curve is sigmoidal within a narrow range of SDS concentrations (0.04 to 0.02%). Size exclusion chromatography and chemical crosslinking revealed that the alpha-helical form is dimeric, while the beta-sheet rich form is tetra- or oligomeric. Both the alpha-helical and beta sheet rich intermediates are soluble and stable. Thus, they should be accessible to further structural and mechanistic studies. At 0.01% SDS, the oligomeric intermediates aggregated into large, insoluble structures as observed by fluorescence correlation spectroscopy. Our results are discussed with respect to the mechanism of PrP(Sc) formation and the propagation of prions. PMID- 11405233 TI - Local variability of the phosphoglycerate kinase-triosephosphate isomerase fusion protein from Thermotoga maritima MSB 8. AB - The pgk-tpi gene locus of Thermotoga maritima encodes both phosphoglycerate kinase (PGK) and a bienzyme complex consisting of a fusion protein of PGK with triosephosphate isomerase (TIM). No separate tpi gene for TIM is present in T. maritima. A frame-shift at the end of the pgk gene has been previously proposed as a mechanism to regulate the expression of the two protein variants [Schurig et al., EMBO J. 14 (1995), 442-451]. Surprisingly, the complete T. maritima genome was found to contain a pgk-tpi sequence not requiring the proposed frameshift mechanism. To clarify the apparent discrepancy, a variety of DNA sequencing techniques were applied, disclosing an anomalous local variability in the pgk-tpi fusion region. The comparison of different DNA samples and the mass spectrometric analysis of the amino acid sequence of the natural fusion protein from T. maritima MSB8 confirmed the local variability of the DNA variants. Since not all peptide masses could be assigned, further variations are conceivable, suggesting an even higher heterogeneity of the T. maritima MSB8 strain. PMID- 11405234 TI - Epigenetics of latent Epstein-Barr virus genomes: high resolution methylation analysis of the bidirectional promoter region of latent membrane protein 1 and 2B genes. AB - We analysed the methylation patterns of CpG dinucleotides in a bidirectional promoter region (LRS, LMP 1 regulatory sequences) of latent Epstein-Barr virus (EBV) genomes using automated fluorescent genomic sequencing after bisulfite induced modification of DNA. Transcripts for two latent membrane proteins, LMP 1 (a transforming protein) and LMP 2B, are initiated in this region in opposite directions. We found that B cell lines and a clone expressing LMP 1 carried EBV genomes with unmethylated or hypomethylated LRS, while highly methylated CpG dinucleotides were present at each position or at discrete sites and within hypermethylated regions in LMP 1 negative cells. Comparison of high resolution methylation maps suggests that CpG methylation-mediated direct interference with binding of nuclear factors LBF 2, 3, 7, AML1/LBF1, LBF5 and LBF6 or methylation of CpGs within an E-box sequence (where activators as well as repressors can bind) is not the major mechanism in silencing of the LMP 1 promoter. Although a role for CpG methylation within binding sites of Sp1 and 3, ATF/CRE and a sis inducible factor (SIF) cannot be excluded, hypermethylation of LRS or regions within LRS in LMP 1 negative cells suggests a role for an indirect mechanism, via methylcytosine binding proteins, in silencing of the LMP 1 promoter. PMID- 11405235 TI - The cytosine N4-methyltransferase M.PvuII also modifies adenine residues. AB - Methylation of DNA occurs at the C5 and N4 positions of cytosine and N6 of adenine. The chemistry of methylation is similar among methyltransferases specific for cytosine-N4 and adenine-N6. Moreover these enzymes have similar structures and active sites. Previously it has been demonstrated that the DNA (adenine-N6)-methyltransferases M.EcoRV, M.EcoRI, E. coli dam and both domains of M.FokI also modify cytosine residues at the N4 position [Jeltsch et al., J. Biol. Chem. 274 (1999), 19538-19544]. Here we show that the cytosine-N4 methyltransferase M.PvuII, which modifies the second cytosine in CAGCTG sequences, also methylates adenine residues in CAGATG/CAGCTG substrates in which the target cytosine is replaced by adenine in one strand of the recognition sequence. Therefore, adenine-N6 and cytosine-N4 methyltransferases have overlapping target base specificities. These results demonstrate that the target base recognition by N-specific DNA methyltransferases is relaxed in many cases. Furthermore, it shows that the catalytic mechanisms of adenine-N6 and cytosine-N4 methyltransferases are very similar. PMID- 11405236 TI - Expression of the human Menkes ATPase in Xenopus laevis oocytes. AB - Menkes disease is an X-linked disorder of copper metabolism that is usually fatal. The affected gene has recently been cloned and encodes one of the two human copper ATPases. If the Menkes ATPase is defective, copper is trapped in the intestinal mucosa, leading to systemic copper deficiency. In order to study copper transport by this ATPase and the effects of disease mutations on its function, we developed a Xenopus laevis oocyte expression system. Wild-type Menkes ATPase cDNA and a fusion of this gene with the green fluorescent protein (GFP) gene was transcribed in vitro and the mRNA injected into oocytes. Expression in oocytes was analyzed by Western blotting and fluorescence microscopy. The Menkes ATPase-GFP chimera appeared to localize primarily to the plasma membrane as assessed by confocal microscopy. This system should thus provide an interesting new tool to study the function of the Menkes ATPase. PMID- 11405237 TI - Biofilms: sensing and signaling. AB - Biofilms are a community of surface-attached microorganisms that can have far reaching effects. Biofilms are costly to industry and affect human health in a variety of ways. Research is only now beginning to discern the complexities of biofilm formation. PMID- 11405238 TI - Differential effects of the novel non-peptidic opioid 4-tyrosylamido-6-benzyl 1,2,3,4 tetrahydroquinoline (CGPM-9) on in vitro rat t lymphocyte and macrophage functions. AB - Opioid receptors have been reported on immune cells of several species and shown to subserve effector functions of these cell types. Mu-selective opioid agonists such as morphine are immunosuppressive, whereas certain delta-opioid receptor selective agonists have been associated with immunopotentiation. We have previously shown that intracerebroventricular administration of the non-peptidic delta-opioid receptor agonists did not alter certain parameters of immunocompetence. In this study, we evaluated the in vitro effects of the novel non-peptidic opioid 4-tyrosylamido-6-benzyl-1,2,3,4 tetrahydroquinoline (CGPM-9) on lymphocyte and macrophage functions. We demonstrated that CGPM-9 enhanced rat thymic lymphocyte proliferative response to concanavalin A (2.85- to 5.5-fold increases), and suppressed LPS-induced nitric oxide (67 to 72 percent reduction) and TNF-alpha production (46 percent reduction) by peritoneal macrophages, compared with untreated control. The mu-opioid receptor selective antagonist CTOP used at equimolar doses, significantly suppressed the effect of CGPM-9 on lymphocyte and macrophage functions (CTOP alone did not show any effect on lymphocyte or macrophage functions). In summary, CGPM-9 activated thymic lymphocyte proliferation and suppressed macrophage functions by acting at mu opioid receptors. This suggests that opioid receptors on immunocytes may be coupled to different signaling pathways depending on the cell type and effector function being analyzed. The mechanism (s) associated with the differential effect of CGPM-9 on these immune cells remains to be elucidated. The pharmacotherapeutic potential for compounds such as CGPM-9 which potentiate T lymphocyte proliferation and suppress production of macrophage-derived inflammatory cytokines is substantial in research and clinical medicine. PMID- 11405239 TI - Preconditioning in the absence or presence of sustained ischemia modulates myocardial Cx43 protein levels and gap junction distribution. AB - In the heart, brief repeated episodes of ischemia prior to a sustained occlusion (ischemic preconditioning; PC) significantly delay the onset of necrosis and arrhythmogenesis. Ischemia has been reported to influence gap junction organization and connexin43 (Cx43) content, but whether PC affects these structures is not known. We investigated the effect of PC (2 cycles of 5-min ischemia plus 10-min reperfusion) followed by prolonged reperfusion without concomitant regional coronary occlusion on the myocardial Cx43 content and its spatial distribution in rabbit hearts. We also compared the effect of sustained ischemia with or without PC on Cx43 spatial distribution. In experiments with PC only, there was an initial decrease in Cx43 levels within the ischemic zone followed by a progressive increase after 48 h reperfusion. End-to-end immunolabeling of Cx43 was augmented in the ischemic region between 24 and 48 h reperfusion; labeling was not uniquely confined to myocyte abutments, but was also dispersed along the sarcolemma. Cx43 immunolabelling was more intense and diffuse in hearts subjected to PC before sustained coronary occlusion (compared to non-PC). These data indicate that gap junctions are significantly altered during brief episodes of ischemia. Reorganization of the gap junction complex could contribute to PC-mediated reductions in cardiac arrhythmias. PMID- 11405240 TI - Skeletal sensitivity to dietary calcium deficiency is increased in the female compared with the male rat. AB - We investigated potential sex differences in bone resorption and the conservation of whole body bone mass in 24-week-old Sprague-Dawley rats maintained on a 1.0% calcium diet and then fed diets containing 0.02, 0.5, 1.0, or 1.75% calcium for 31 days. Lowering dietary calcium from 1.00% to 0.02% doubled whole skeleton bone resorption (urinary 3H-tetracycline loss). Female rats were more sensitive to calcium stress, exhibiting the maximal resorptive response when fed the 0.5% calcium diet, whereas the 0.02% calcium diet was required to elicit this response in males. Despite the evidence of increased bone resorption, whole skeleton mass was unchanged in females and was significantly increased in males, indicating that switching to even the 0.02% calcium diet did not result in an overt loss of total body bone mass. Compared with controls, the skeleton mass of females (97+/ 1.4%) maintained on the 0.02% calcium diet was significantly lower than males (107+/-2.4%), again suggesting a greater impact of calcium deficiency in females. The calculation of the average percentage growth of selected individual bones in male rats indicated a proportional increase in bone mass between the axial and appendicular skeleton of approximately +4% and +18% in animals maintained on 0.02 and 1.75% diets, respectively. By comparison, female rats consuming the 0.02% calcium diet showed an average 14% loss in axial bone and 7.5% gain in appendicular bone mass. The results indicate increased sensitivity to dietary calcium deficiency in female rats which involves a significant loss in axial bone mass not observed in male rats maintained under similar dietary conditions. PMID- 11405241 TI - Changes in skeletal muscle in males and females following endurance training. AB - Gender differences in substrate selection have been reported during endurance exercise. To date, no studies have looked at muscle enzyme adaptations following endurance exercise training in both genders. We investigated the effect of a 7 week endurance exercise training program on the activity of beta-oxidation, tricarboxylic acid cycle and electron transport chain enzymes, and fiber type distribution in males and females. Training resulted in an increase in VO2peak, for both males and females of 17% and 22%, respectively (P < 0.001). The following muscle enzyme activities increased similarly in both genders: 3-beta hydroxyacyl CoA dehydrogenase (38%), citrate synthase (41%), succinate-cytochrome c oxidoreductase (41%), and cytochrome c oxidase (COX; 26%). The increase in COX activity was correlated (R2 = 0.52, P < 0.05) with the increase in VO2peak/fat free mass. Fiber area, size, and % area were not affected by training for either gender, however, males had larger Type II fibers (P < 0.05) and females had a greater Type I fiber % area (P < 0.05). Endurance training resulted in similar increases in skeletal muscle oxidative potential for both males and females. Training did not affect fiber type distribution or size in either gender. PMID- 11405242 TI - Chronotropic response to (+/-)-CGP12177 in right atria of stressed rats. AB - Foot-shock stress changes the sensitivity of the rat right atria to beta1- and beta2-adrenoceptor (AR) agonists. We investigated whether the same stress protocol also changes the atrial sensitivity to the non conventional agonist, (+/ )-CGP12177. Concentration-response curves to (+/-)-CGP12177, a beta1- and beta2 adrenoceptor antagonist with agonist properties at the putative beta4 adrenoceptors, were obtained in the absence and presence of propranolol (200 nM or 2 microM), CGP20712A 10 nM plus ICI118,551 50 nM, or CGP20712A (1 microM or 3 microM), in right atria from rats submitted to three daily foot-shock sessions (120 mA pulses of 1.0 s duration applied at random intervals of 5-25 s over 30 min) and killed after the third session. The pD2 for (+/-)-CGP12177 was not influenced by foot-shock stress. The stimulant effect of (+/-)-CGP12177 was resistant to blockade by 200 nM and 2 microM (+/-)-propranolol, and to combined blockade by CGP20712A and IC1118,551. However, in right atria from stressed rats given 200 nM propranolol, the concentration-response curve to the agonist was shifted 2.0-fold to the right. CGP20712A shifted the concentration-response curve to (+/-)-CGP12177 to the right by 4.6- (1 microM) and 19-fold (3 microM) in atria of control rats, and by 2.2- (1 microM) and 43-fold (3 microM) in atria of stressed rats. Maximum response to CGP12177 was not affected by propranolol or CGP20712A in concentrations ranging from 0.1 nM to 10 microM. These results show that the chronotropic effect of (+/-)-CGP12177 is mediated by atypical beta4 adrenoceptors. In constrast with to beta1-and (or) beta2-AR, this receptor is resistant to the effects of foot-shock stress, suggesting that the putative beta4 AR is a different receptor from a low affinity state of beta1-adrenoceptor, as previously proposed, unless both proposed isoforms of beta1-adrenoceptor show independent stress-induced behavior. PMID- 11405243 TI - Estrogen effect on post-exercise skeletal muscle neutrophil infiltration and calpain activity. AB - We hypothesized that estrogen administration would attenuate skeletal muscle neutrophil infiltration, indices of muscle membrane disruption, and muscle calpain activity shortly after the termination of exercise. Ovariectomized female rats were implanted with either an estogen pellet (25 mg beta-estradiol) or a placebo pellet. Two weeks postimplant, animals were killed either at rest or 1 h after running exercise (60 min at 21 m x min(-1), 12% grade). The 4 experimental groups (n = 12) used were: unexercised placebo (UP), unexercised estrogen (UE), exercised placebo (EP), and exercised estrogen (EE). Blood samples were analyzed for creatine kinase (CK) activity and estradiol content. Plantaris and gastrocnemius muscles were removed and histochemical determination of neutrophil content or biochemical determination of myeloperoxidase (MPO), glucose-6 phosphate dehydrogenase (G6PD), and calpain-like activity determined. Estrogen supplemented animals had 10-20-fold higher circulating estradiol levels than placebo animals. EP animals had significantly higher (P < 0.05) circulating CK activities than EE or unexercised animals. Muscle neutrophil concentrations were significantly (P < 0.01) elevated in EP and EE groups compared with unexercised controls, with EP muscle neutrophil levels also being over 60% greater (P < 0.05) than in EE animals. EP animals also had higher (P < 0.05) muscle MPO activities than unexercised or EE animals. Muscle G6PD activities were not significantly different between any groups. Muscle caplain-like activities were 80% higher (P < 0.01) in EP animals than EE animals with calpain-like activities in EE animals similar to unexercised groups. These results indicate that estrogen supplementation in ovariectomized rats attenuated 1-h post-exercise serum CK activities, muscle neutrophil infiltration, MPO activities, and calpain-like activities when compared with exercised, unsupplemented animals. This supports the possibility of a relationship between estrogen, calpain dependent production of neutrophil chemo-attractant peptides, and 1-h post-exercise skeletal muscle neutrophil infiltration. PMID- 11405244 TI - Relationship between extra and intracellular sources of calcium and the contractile effect of thiopental in rat aorta. AB - To evaluate the relationship between the vasocontractile effect of thiopental and the extra and intracellular sources of Ca2+, we analyzed both the contractile effect of the barbiturate on rat aortic rings and its ability to modify the intracellular calcium concentration in cultured rat aorta smooth muscle cells. Thiopental (10-310 microg/mL) contracted aortic rings only in the presence of extracellular Ca2+, and this effect was not blocked by verapamil or diltiazem. On the contrary, Ca2+ (0.1-3.1 mM) evoked contractions only when thiopental (100 microg/mL) was present. Although in calcium-free solution thiopental (100 microg/mL) did not contract aortic rings, it abolished the contractile effect of either phenylephrine (10(-6) M) or caffeine (10 mM). Finally, thiopental augmented the intracellular calcium concentration in cultured smooth muscle cells incubated either in the presence or absence of calcium. In conclusion, thiopental's vasocontractile effect depends on extracellular calcium influx, which is independent of L-calcium channels. The increase in intracellular Ca2+ concentration elicited by thiopental in Ca2+-free solution and its ability to block the effect of phenylephrine and caffeine suggest that this barbiturate can deplete intracellular pools of calcium. Therefore, the calcium entry pathway associated with the contractile effect of thiopental may correspond to the capacitative calcium entry model. PMID- 11405245 TI - Study of acute pharmacologic effects of prolactin on calcium and water transport in the rat colon by an in vivo perfusion technique. AB - We investigated the acute effect of intraperitoneally administered prolactin on calcium and water transport in colon of sexually mature female Wistar rats using an in vivo perfusion technique. Test solution containing (in mM) NaCl, 100; KCl, 4.7; MgSO4, 1.2; CaCl2, 20; D-glucose, 11; sodium ferrocyanide (Na4Fe(CN)6), an index of net water transport, 20; and 0.7 (microCi 45CaCl2 (1 Ci = 37 GBq) was perfused througth the 8-cm colonic loop for 60 min at perfusion rates of 0.5 or 1.0 mL x min(-1). Calcium and water transport was also studied under a no flow condition to stimulate the condition often found in the colon by in vivo ligated colonic loop for 30 min. Control results showed no correlation between calcium transport and water flux. Flow of luminal solution at 0.5 and 1.0 mL x min(-1) was found to reverse net calcium absorption from 0.04+/-0.01 nmol x g(-1) dry weight x h(-1) to net calcium secretion of 0.04+/-0.04 and 0.9+/-0.02 nmol x g( 1) dry weight x h(-1), respectively. Neither 0.4, 0.6, nor 1.0 mg x kg(-1) prolactin had any effect on calcium fluxes in the colon. On the other hand, at a perfusion rate of 1 mL x min(-1), 0.4 mg x kg(-1) prolactin significantly decreased net water absorption from 3.86+/-0.90 to 0.88+/-0.64 mL x g(-1) dry weight x h(-1) (P < 0.001), and the higher doses of 0.6 and 1.0 mg x kg(-1) prolactin reversed net water absorption to net water secretion of 2.20+/-0.63 and 2.33+/-0.89 mL x g(-1) dry weight x h(-1), respectively (P < 0.001). The stimulatory effect of prolactin on water transport was completely abolished by reducing the perfusion rate from 1.0 mL x min(-1) to zero. The stimulatory effect of prolactin on water secretion at perfusion rate of 1.0 mL x min(-1) was also abolished when luminal [Na+] was reduced from 180 to 80 mM. We concluded that, unlike in the small intestine, calcium fluxes in the colon are not related to water transport and did not respond at all to prolactin. Water transport, on the other hand, was reversed from net absorption to secretion by prolactin. We propose that this prolactin-induced water secretion is probably mediated by recycling of luminal sodium in the vicinity of tight junctions. PMID- 11405246 TI - Modulation of AMPA receptors by a novel organic nitrate. AB - Positive modulators of alpha-amino-3-hydroxy-5-methylisoxazole-4-propionic acid (AMPA) channels reduce desensitization and alter their gating kinetics. We have discovered a novel compound nitric oxide-mimetic that similarly modulates the AMPA receptor by reducing desensitization. This, designated GT-005, belongs to the organic nitrate family that includes the nitrovasodilator nitroglycerine. In acutely isolated hippocampal neurons, GT-005 enhanced kainate (100 microM)-evoked currents with an EC50 of 1.7+/-0.2 mM and a 176+/-10% maximal increase in the steady-state current response. Similar results were found in cultured hippocampal neurons (EC50 of 1.3+/-0.2 mM and a maximal 83+/-14% increase in the steady-state current response). GT-005 reduced the desensitization of glutamate-evoked currents and slowed the onset of desensitization. This compound also increased the rate of recovery from the desensitized state. With respect to alteration of the excitatory synaptic transmission, GT-005 delayed the decay and increased the frequency of spontaneous miniature excitatory postsynaptic currents (mepsc) recorded in cultured hippocampal neurons. PMID- 11405247 TI - Prolactin directly stimulates transcellular active calcium transport in the duodenum of female rats. AB - Prolactin has been postulated to be a novel calcium-regulating hormone during pregnancy and lactation. It stimulates both passive and active duodenal calcium transport in several experimental models. Our study was performed on sexually mature female Wistar rats (200-250 g) to study the direct action of prolactin on calcium transport in the duodenum using the Ussing chamber technique. To evaluate the effect of prolactin on total calcium transport in the duodenum, we intraperitoneally injected rats with 0.4, 0.6, and 0.8 mg/kg prolactin. The total calcium transport was divided into voltage-dependent, solvent drag-induced, and transcellular active fluxes by applying short-circuit current and by mucosal glucose replacement with mannitol. The effect of prolactin on each flux was studied separately. Finally, to evaluate the direct action of prolactin on duodenal transcellular active flux, we directly exposed duodenal segments to prolactin that had been added to the serosal solution with or without calcium transport inhibitors. We found that 0.6 and 0.8 mg/kg prolactin ip significantly increased the total mucosa-to-serosa calcium flux from the control value (nmol x hr(-1) x cm(-2)) of 34.53+/-6.81 to 68.07+/-13.53 (P < 0.05) and 84.43+/-19.72 (P < 0.01), respectively. Prolactin also enhanced the solvent drag-induced calcium flux and transcellular active calcium flux, but not the voltage-dependent calcium flux. The duodenal segments directly exposed to 200, 400, and 800 ng/mL prolactin showed a significant increase in the transcellular active calcium absorption in a dose-dependent manner, i.e., from the control value (nmol x hr(-1) x cm(-2)) of 2.94+/-0.47 to 5.45+/-0.97 (P < 0.01), 8.09+/-0.52 (P < 0.001), and 18.42+/-2.92 (P < 0.001), respectively. Its direct action was inhibited by mucosal exposure to 50 microM lanthanum chloride, a calcium transporter protein competitor, and serosal exposure to 0.1 mM trifluoperazine, a Ca2+-ATPase inhibitor. These studies demonstrate that the duodenum is a target organ of prolactin, which enhances transcellular active calcium transport. PMID- 11405248 TI - Proteinase-activated receptor 4 (PAR4): activation and inhibition of rat platelet aggregation by PAR4-derived peptides. AB - We studied the actions of receptor-activating peptide analogues (PAR4APs), modeled on the proteolytically-revealed tethered ligand sequence of murine proteinase-activated receptor-4 (PAR4), in a rat platelet aggregation assay. The PAR4APs GYPGKF-NH2 (GY-NH2) and AYPGKF-NH2 (AY-NH2) were able to cause aggregation with EC50 values of about 40 microM and 15 microM, respectively. The reverse human PAR4 sequence (VQGPYG-NH2, YG-NH2) and the PAR1AP SFLLR-NH2, did not cause aggregation. In contrast, trans-cinnamoyl-YPGKF-NH2 (tcY-NH2) did not cause aggregation but blocked aggregation caused by GY-NH2, AY-NH2, and thrombin without affecting ADP-mediated aggregation. We conclude that in contrast to the PAR1AP, the PAR4APs GY-NH2 and AY-NH2 activate rat platelets via a PAR4-related receptor and that peptide analogues modeled on the PAR4 tethered activating sequence can serve as useful agonist and antagonist probes for assessing the consequence of activating PAR4 either by PAR4APs or thrombin in rat tissue preparations. PMID- 11405249 TI - Round window membrane delivery of L-methionine provides protection from cisplatin ototoxicity without compromising chemotherapeutic efficacy. AB - Cisplatin (cis-diamminedichloroplatinum(II) (CDDP)) is a widely used, highly effective, oncolytic agent that has serious ototoxic side-effects. To test the effectiveness of local delivery, of L-methionine (L-Met) as an otoprotective agent against CDDP ototoxicity, we used a rat model of a highly metastatic breast cancer tumor, i.e. Fisher 344 rats implanted with MTLn3 breast cancer cells. Four experimental groups were evaluated--I: untreated; II: CDDP-treated (three dosages); III: systemically-delivered L-Met + CDDP-treated; IV: locally delivered L-Met + CDDP-treated. The integrity of the outer hair cells (OHCs) was determined using scanning electron microscopy (SEM); hearing was assessed by recording auditory brainstem responses (ABRs) at multiple frequencies. The chemotherapeutic effectiveness of CDDP was quantified by measuring changes in tumor mass and the presence of tumor metastasis. L-Met provided otoprotection of the OHCs against CDDP toxicity in the cochleae of rats following either systemic (III) or local (IV) administration. The ABRs were unchanged in each of the L-Met protection Groups (III and IV) and in the untreated animals of Group I. Treatment with CDDP only (II) induced significant hearing losses at both 16 and 18 kHz when compared to ABRs of untreated rats(I). CDDP was effective in controlling the MTLn3 initiated breast cancer tumors in the CDDP-treated (II) and the local L-Met protection, CDDP-treated (IV) Groups. In contrast, the tumors in the systemic L Met protection, CDDP-treated Group (III) were not controlled by the CDDP treatment regime. This study demonstrates that local delivery of L-Met to the scala tympani of the cochlea via the round window membrane (IV) provides effective protection against CDDP ototoxicity without compromising its ability to control a highly metastatic form of cancer. PMID- 11405250 TI - Effects of repeated hydrogen sulphide (H2S) exposure on learning and memory in the adult rat. AB - The effects of repeated exposure (125 ppm) of hydrogen sulphide (H2S) on learning and memory in the rat were investigated. A 16-arm radial arm maze (RAM) was used to examine neurobehavioural functioning in a series of three experiments. Experiment 1 involved training animals on a complex spatial maze task, prior to a 5-week period of exposure to H2S or a control gas mixture. Rats were tested for maze retention after each 5-day exposure period. It was determined that repeated H2S exposure had no effect on memory for a previously learned spatial task. Experiment 2 was conducted to determine whether H2S interferes with the acquisition of a novel spatial task. Naive animals received daily maze training and exposure (H2S or control) sessions over an extended 11-week period (48 sessions). The results indicated that the groups were comparable on four of five measures of maze performance. H2S animals were impaired in their ability to find all of the reinforcers prior to the end of a trial, suggesting that H2S had an effect on performance rate, but not acquisition of the maze task. Finally, Experiment 3 was conducted to determine what role proactive interference might play in H2S-related brain impairment. Animals from the preceding experiment were trained on a new reversed contingency maze task. H2S animals made more overall arm entries than controls, suggesting that H2S may impair learning by increasing the animals' susceptibility to interference from irrelevant stimuli. The prefrontal cortex was discussed as a potential target site of H2S. The pathophysiological mechanisms underlying the effect of H2S on normal brain function have yet to be identified. PMID- 11405251 TI - A comparative study of binding sites for diisopropyl phosphorofluoridate in membrane and cytosol preparations from spinal cord and brain of hens. AB - Biochemical events in the initiation of organophosphorus induced delayed neurotoxicity (OPIDN) are not well understood. To find new putative target(s) for OPIDN, we investigated the biochemical and pharmacological characteristics of [3H] diisopropyl phosphorofluoridate (DFP) binding to membrane and cytosol preparations from the brain and spinal cord of hens in vitro. [3H]DFP binding to both preparations was determined by the specific binding obtained by subtracting non-specific binding from total binding. The specific binding sites of [3H]DFP were found not only on membrane but also in cytosol. Kd values were higher and Bmax values were lower in cytosol than in membrane. Moreover, the Kd values in both membrane and cytosol preparations from spinal cord were lower than those of brain. The Bmax values in membrane and cytosol were similar between brain and spinal cord. The specific binding to both preparations was markedly displaced by unlabeled DFP. The specific binding of DFP to the membrane was highly or partly displaced by organophosphorus compounds (OPs) or a carbamate, respectively. However, both the OPs and the carbamate had considerably weaker blocking effects on the specific binding of DFP to cytosol. None of the compounds known to interact with neuropathy target esterase (NTE) had a strong blocking effect on the specific binding of DFP to either membrane or cytosol. These results show that the specific binding of DFP to the membrane may be binding with cholinesterase (ChE). However, cytosol, especially in spinal cord, may have DFP binding sites other than ChE and NTE. PMID- 11405252 TI - Correlation of binding sites for diisopropyl phosphorofluoridate with cholinesterase and neuropathy target esterase in membrane and cytosol preparations from hen. AB - To find new putative target(s) for organophosphorus induced delayed neurotoxicity (OPIDN), we investigated the biochemical and pharmacological characteristics of [3H] diisopropyl phosphorofluoridate (DFP) binding to membrane and cytosol preparations from the brain and spinal cord of hens. Specific [3H]DFP binding was determined by subtracting non-specific binding from total binding. The binding sites of [3H]DFP, an organophosphate that induces OPIDN, were found not only on membrane but also in cytosol. Reduction of subsequent ex vivo specific [3H]DFP binding by in vivo pretreatment with unlabeled DFP was found in cytosol, not membrane. The reduced binding lasted to the onset of OPIDN, especially in spinal cord. These results suggest that the specific DFP binding sites in cytosol, rather than on membrane, are the most important with regard to the initiation of OPIDN. Inhibitors of cholinesterase (ChE) and neuropathy target esterase (NTE) other than DFP did not affect specific [3H]DFP binding to either membranes or cytosol in vivo. Additionally, inhibition of the activities of these esterases by these compounds was not consistent with either the degree of inhibition of the [3H]DFP binding or a time-dependent manner of OPIDN. These results suggest that DFP binding site(s) involved in the initiation of OPIDN may be different from the active sites of ChE and NTE. PMID- 11405253 TI - Gamma-diketone peripheral neuropathy III. Neurofilament gene expression. AB - Evidence suggests the morphologic hallmark of gamma-diketone neuropathy is axon atrophy and that this effect is associated with reduced neurofilament (NF) subunit protein content (Toxicol Appl Pharmacol 2000;165:141-7). To investigate the mechanism of diminished NF content, subunit (NF-L, -M and -H) gene expression was quantified in dorsal root ganglion (DRG) of slightly affected and moderately intoxicated groups of rats exposed to 2,5-hexanedione (HD) at one of three daily dosing rates (175, 250 and 400 mg/kg per day). Results show that sensory ganglia from slightly affected rats exhibited no changes in gene expression, whereas at a moderate level of neurotoxicity, each dosing protocol was associated with small but significant reductions (approximately 20%) in mean NF subunit mRNA. This was not a generalized effect on expression of cytoskeletal components in sensory ganglia since tubulin message levels were not affected. Although the observed reduction in NF gene expression might be related to diminished levels of subunit proteins in peripheral nerve, the actual contribution is likely to be minimal. The magnitude of effect was small and did not correspond to the dose-rate dependent effect of HD on respective isotype proteins. The mechanism of gamma diketone-induced axon atrophy is unknown but might involve local changes in axonal NF phosphorylation and degradation. PMID- 11405254 TI - Intracerebral administration of 2,4-diclorophenoxyacetic acid induces behavioral and neurochemical alterations in the rat brain. AB - Although, the mechanism of 2,4-dichlorophenoxyacetic acid (2,4-D) neurotoxicity remains unknown, the monoaminergic system appears to mediate some of its effects in rats as we previously reported. In this study; we examined the 2,4-D effects on locomotor activity, circling behavior and monoamine levels after the injection into the basal ganglia of male adult rats. These effects were compared with those induced after selective lesions of dopaminergic neurons with 6-hydroxydopamine (6 OHDA). 2,4-D-injected into one striatum (100 microg/rat) produced a marked depression in locomotor activity and elicited a moderate circling towards the ipsilateral side at 6 and 24 h postinjection. These behavioral changes were accompanied by a decrease and an increase of serotonin (5-HT) and homovanillic acid (HVA) levels, respectively. 2,4-D administration (100 microg/rat) into the nucleus accumbens, induced similar behavioral and neurochemical patterns to the intrastriatal 2,4-D injection, although rats did not present notorious turning. When 2,4-D was injected into one medial forebrain bundle (MFB, 50 microg/rat), animals presented ipsilateral circling, while locomotor activity was unchanged at 3 and 7 days post-injection. These last rats also exhibited diminished levels of striatal 5-HT, dopamine (DA) and their metabolites without changes in the substantia nigra (SN). Animals sacrificed 3 and 7 days after a 6-OHDA injection into one of the MFB, presented progressive depletion of dopamine in striatum and SN. 2,4-D as well as 6-OHDA-treated rats into one of the MFB were challenged with low dose (0.05 mg/kg s.c.) of apomorphine (only at 7 days post-injection) to evaluate a possible DA-receptor supersensitivity. Only 6-OHDA treated rats showing a vigorous contralateral rotation activity. These results indicate that 2,4-D induced a regionally-specific neurotoxicity in the basal ganglia of rats. The neurotoxic effects of 2,4-D on basal ganglia by interacting with the monoaminergic system depended not only on the exact location of the 2,4-D injection, but also on the dose and time period of post-injection. Toxicity produced by 2,4-D appears to be different in monoaminergic terminals, axonal fibers, and cell bodies. PMID- 11405255 TI - Acute neurobehavioral effects in rats from exposure to HFC 134a or CFC 12. AB - 1,1,1,2-Tetrafluoroethane (HFC 134a), a chlorine-free hydrofluoroalkane, is internationally replacing billions of pounds of dichlorodifluoromethane (CFC 12) for coolant, refrigerant and aerosol propellant applications. The ALC50 for HFC 134a in rats is 567,000 ppm for 4 h; its potential for cardiac epinephrine sensitization in beagle dogs is acceptable (75,000 ppm); and its capacity to induce carcinogenicity or developmental disorders in animals is minimal. HFC 134a, with a serum half life estimated at 4-11 min, has been accepted for use as a propellant in metered-dose inhalant products, implying a low human toxicity risk from periodic brief exposures. There has been little published human or animal research evaluating possible neurobehavioral toxicity from longer HFC 134a exposures, as may be expected to occur in operational scenarios. In this study, male Wistar rats were exposed to various concentrations of HFC 134a or CFC 12 for up to 30 min while performing in either a rotarod/motorized running wheel apparatus or in an operant chamber The relative neurobehavioral toxicity of CFC 12 and its ozone-depleting substance replacement HFC 134a was assessed by comparing both gross motor system incapacitation and more subtle changes in ability to perform an operant discrimination task. It was shown that exposure to HFC 134a or CFC 12 concentrations from 40,000 to 470,000 ppm, for up to 30 min, induced neurobehavioral deficits in every subject, ranging from reduced operant efficiency to apparent anesthesia. For neurobehavioral endpoints examined in these experiments, HFC 134a inhalation was shown to induce deficits more rapidly, and at lower concentrations when compared to CFC 12 exposure. PMID- 11405256 TI - Neuropsychological effects of low mercury vapor exposure in chloralkali workers. AB - Neuropsychological effects were examined in 47 mercury vapor exposed male chloralkali workers with current low concentrations of urinary mercury (mean U-Hg 5.9 nmol/mmol creatinine (Cr)). Their average duration of exposure was 13.3 years, and the calculated mean concentration of U-Hg was 9.0 nmol Hg/mmol Cr per year (exposure intensity) during their time of exposure. They were compared with 47 age-matched male referents in a cross-sectional study. The two groups were not statistically significantly different with respect to neuropsychological test performance or number of self-reported subjective symptoms. The test results of the Static Steadiness Test, which assesses tremor, were not associated with exposure to mercury vapor. However current smokers had more hand tremor than non smokers. Statistically significant associations were found between indices of current exposure (the concentration of inorganic mercury in whole blood) and the results of the WAIS Digit Symbol Test and the Benton Visual Retention Test (number of correct responses). This could indicate a small effect of current exposure on visuomotor/psychomotor speed and attention, and immediate visual memory. Whether the association found between the historical exposure intensity and the Digit Symbol Test results may represent long-term consequences of exposure cannot be determined in this study. PMID- 11405257 TI - Postnatal inorganic lead exposure decreases the number of spontaneously active midbrain dopamine neurons in the rat. AB - This study examined the effect of lead (Pb) exposure during postnatal development on the electrophysiological activity of midbrain dopamine (DA)-containing neurons. Single-cell electrophysiological recordings were made in the substantia nigra (SN) and ventral tegmental area (VTA) of chloral hydrate anesthetized rats. In this post-weaning exposure protocol 22-day-old male Sprague-Dawley rats were exposed to Pb- (100, 250, and 500 ppm) or Na-acetate in the drinking water for a period ranging from 3 to 6 weeks. Animals were exposed up to the day of electrophysiological recording. One Pb- and one Na-treated animal were recorded each experimental day. The post-weaning exposure protocol used in this study resulted in a significant Pb-dependent decrease in the number of spontaneously active DA neurons at the time of electrophysiological recording. Analysis of covariance, using duration of exposure as the covariate (i.e. 3, 4, 5, or 6 weeks), did not indicate that there was a consistent relationship between exposure duration and the number of spontaneously active DA neurons. However, the effect of Pb was dependent on the level of Pb exposure through the drinking water. At the 250 and 500 ppm level of exposure, Pb produced a significant decrease in the number of spontaneously active DA neurons in both anatomical regions. The number of active DA neurons was not significantly affected by the 100 ppm Pb treatment over the 3-6 weeks exposure period. The average discharge rate, and the percentage of spontaneously active DA neurons classified as having discharge patterns with bursts (i.e. 'bursting DA neurons'), was not changed at any of the three levels of Pb exposure. Based on results obtained from electrophysiological studies, the effect of selected Pb exposure levels, 250 and 500 ppm, were examined during the postnatal period using tyrosine hydroxylase (TH) immuno-histochemistry to determine if Pb affects the survival of dopamine neurons within SN and VTA. TH immuno-histochemical studies revealed that the reduction in the number of spontaneously active DA neurons in animals treated with 250 and 500 ppm Pb was probably not related to the physical loss of cells (e.g. necrosis or apoptosis). PMID- 11405258 TI - Depletion of energy metabolites following acetylcholinesterase inhibitor-induced status epilepticus: protection by antioxidants. AB - Status epilepticus (SE)-induced neuronal injury may involve excitotoxicity, energy impairment and increased generation of reactive oxygen species (ROS). Potential treatment therefore should consider agents that protect mitochondrial function and ROS scavengers. In the present study, we examined whether the spin trapping agent N-tertbutyl-alpha-phenylnitrone (PBN) and the antioxidant vitamin E (DL-alpha-tocopherol) protect levels of high-energy phosphates during SE. In rats, SE was induced by either of two inhibitors of acetylcholinesterase (AChE), the organophosphate diisopropylphosphorofluoridate (DFP, 1.25 mg/kg, sc)- or the carbamate carbofuran (1.25 mg/kg, sc). Rats were sacrificed 1 h or 3 days after onset of seizures by head-focused microwave (power, 10 kW; duration 1.7 s) and levels of the energy-rich phosphates adenosine triphosphate (ATP) and phosphocreatine (PCr) and their metabolites adenosine diphosphate (ADP) and adenosine monophosphate (AMP), and creatine (Cr), respectively, were determined in the cortex, amygdala and hippocampus. Within 1 h of seizure activity, marked declines were seen in ATP (34-60%) and PCr (25-52%). Total adenine nucleotides (TAN = ATP + ADP + AMP) and total creatine compounds (TCC = PCr + Cr) were also reduced (TAN 38-60% and TCC 25-47%). No changes in ATP/AMP ratio were seen. Three days after the onset of seizures, recovery of ATP and PCr was significant in the amygdala and hippocampus, but not in the cortex. Pretreatment of rats with PBN (200 mg/kg, ip, in a single dose), 30 min before DFP or carbofuran administration, prevented induced seizures and partially prevented depletion of high-energy phosphates. Pretreatment with the natural antioxidant vitamin E (100 mg/kg, ip/day for 3 days), partially prevented loss of high energy phosphates without affecting seizures. In controls, citrulline, a product of nitric oxide synthesis, was found to be highest in the amygdala, followed by hippocampus, and lowest in the cortex. DFP- or carbofuran-induced seizures caused elevation of citrulline levels seven- to eight-fold in the cortex and three- to four-fold in the amygdala and hippocampus. These results suggest a close relationship between SE, excitotoxicity and energy metabolism. The involvement of oxidative stress is supported by the findings that DFP and carbofuran trigger an excessive nitric oxide (NO) production in the seizure relevant regions of the brain. PMID- 11405259 TI - Effect of antioxidants on L-glutamate and N-methyl-4-phenylpyridinium ion induced neurotoxicity in PC12 cells. AB - The neuropathology associated with Parkinson's disease within and around the substantia nigra is thought to involve excessive production of free radicals, dopamine autoxidation, defects in the expression of glutathione peroxidase, attenuated levels of reduced glutathione, altered calcium homeostasis, excitotoxicity and genetic defects in mitochondrial complex I activity. While the neurotoxic mechanisms are vastly different for excitotoxins and N-methyl-4 phenylpyridinium ion (MPP+), both are thought to involve free radical production, compromised mitochondrial activity and excessive lipid peroxidation. In the present study, several dietary antioxidant compounds, monoamine oxidase inhibitors and ergogenic compounds were examined for protective action against neurotoxicity induced by L-glutamate (15 mM) or MPP+-HCl (5 mM) in a plastic adhering variant of murine pheochromocytoma cells. The results show no significant protective effects exhibited by azulene, (+)-catechin, curcrumin, (-) epigallocatechin gallate, green tea, morin, pygnogenol, silymarin, clove oil, garlic oil or rosemary, extract. Compounds, which were effective in providing protection against L-glutamate-induced cell death, were coenzyme Q-0, coenzyme Q 10, L-deprenyl and N-acetyl-L-cysteine. Compounds, which provided protection against MPP+-HCl toxicity, were allopurinol, coenzyme Q-10, L-deprenyl, N-acetyl L-cysteine and sesame oil. In both models, significant protection was achieved in the presence of coenzyme Q-10, L-deprenyl and N-acetyl-L-cysteine. These results indicate that the mechanism of cell death in both of these toxicity models is most likely not related to the destructive effects of free radicals. PMID- 11405260 TI - The effect of obliquity on the radiographic appearance of the temporomandibular joint in dogs. AB - The temporomandibular joint is formed between the condyloid process of the mandible and the mandibular fossa of the temporal bone. The basic anatomy of this joint was assessed and described in a series of skulls including dolichocephalic, mesaticephalic and brachycephalic breeds. The facial index and rotational angles were measured with the facial index providing a useful method of classifying skull types but the rotational angle being of limited use in assessment of the temporomandibular joint until normal breed values are established. Equipment was designed to allow repeatable positioning of the temporomandibular joint for radiography at a variety of lateral and long axis rotational angles relative to the central x-ray beam. The regions of the joint and anatomic features visualized in each view are demonstrated. 10 degrees rotation was required in either axis to project the joints independently of each other. Lateral rotational angles of 10 to 30 degrees in mesaticephalic and dolichocephalic breeds and 20 to 30 degrees in brachycephalics and long axis rotational views of 10 to 30 degrees depending on the region of interest were considered to be the most useful. PMID- 11405261 TI - Osteosarcoma of the patella with pulmonary metastases in a dog. AB - A 6-year-old neutered male Rottweiler was examined for a progressive right pelvic limb lameness. In radiographs of the right stifle, there was an osteolytic lesion with irregular new bone formation along the cranial aspect of the patella consistent with an aggressive bone lesion. In thoracic radiographs, there were multiple soft tissue nodular opacities throughout the lung fields, consistent with pulmonary metastases. Microscopically, fine needle aspirate samples from the right patella contained pleomorphic spindle cells with cytologic features of osteosarcoma. The presence of pulmonary metastases at the time of initial diagnosis in the dog described herein suggests that osteosarcoma of the patella has the potential for similar aggressive biologic behavior as that seen in dogs with appendicular osteosarcoma. PMID- 11405262 TI - Computed tomographic imaging of infiltrative lipoma in 22 dogs. AB - Twenty two dogs with an infiltrative lipoma had computed tomographic (CT) images acquired to evaluate the extent of local disease. Ten dogs had undergone at least one cytoreductive surgical procedure (range = 1-3; median = 2) prior to imaging. Twenty dogs had measurable disease on CT images; 2 dogs had diffuse disease at a previous surgical site that could not be measured. Tumor volume (n = 20) ranged from 20 to 5,632 cm3 (median = 345 cm3; mean = 996 cm3). None of the dogs had evidence of bone involvement on the CT images; 2 of the 22 dogs had tumors that did not come into direct contact with osseous structures. All dogs with measurable disease had evidence of a fat opacity mass with variable degrees of muscle infiltration. Eleven of 22 dogs were given intravenous contrast medium prior to image acquisition and there was not evidence of enhancement of the infiltrative lipoma in any dog. Based on CT images, tumors were classified as well-defined in 9 dogs, moderately well-defined in 4, not well-defined in 3 and a mix of well-defined and not well-defined in 6 dogs. Tumors tended to be less well defined in regions where the infiltrative lipoma interdigitated with normal body fat. It appears CT imaging allows adequate discrimination of tumor with the caveat that differentiation of normal fat from infiltrative lipoma can be problematic. PMID- 11405263 TI - Computed tomographic anatomy of the canine pancreas. AB - Barium sulfate was administered into the coeliac artery of 5 canine cadavers to allow for contrast computed tomography of the pancreas. Contiguous, 2-mm-thick slices were acquired. Multiplanar and three-dimensional reformatting were performed to clarify the anatomic relationship. After imaging, the cadavers were frozen, cross sections obtained, and plastinated. These were compared to the computed tomography images. Five plain and contrast enhanced computed tomographic series of normal live controls were acquired and evaluated retrospectively. In the study of the canine cadavers the pancreas became opacified and appeared homogenous with irregular contour. In normal live controls, acquiring an image at the end of expiration allowed a detailed view of the pancreatic parenchyma in the non-alterated pancreas, but pancreatic and bile ducts could not be seen. Adjacent to the hepatic hilus the pancreatic body appeared as a dorsoventrally flattened structure bordering on the ventral surface of the portal vein, both in cadavers and normal live controls. The right lobe extended caudodorsally to the right abdominal wall and aligned with the cranial part of the duodenum. The left lobe was adjacent to the gastric body in all dogs although it was separated from the gastric fundus by the dorsal extremity of the spleen in normal live controls. Neither kidney was suitable as an anatomic marker for localization of the pancreas, unlike traditional references in textbooks. We recommended using the portal vein to localize the pancreatic body, the descending duodenum for the right lobe, and the dorsal extremity of the spleen as well as the gastric fundus for the left lobe. PMID- 11405264 TI - Radiographic diagnosis: cartilaginous exostoses in a dog. AB - A 6 month-old dog was examined for progressive paraparesis. On physical examination bony malformations were palpated over the cranial lumbar vertebral bodies and on the left metatarsal bone. Neuroanatomic lesion localization for the paraparesis was a T3-L3 spinal cord lesion. Radiographs confirmed bony masses at L1-L2 and on the left 3rd metatarsal bone. Magnetic resonance imaging was performed from T3-L3. Severe spinal cord compression was identified at L1-L2. Surgical decompression and biopsy confirmed the mass to be cartilaginous exostoses. This paper is an example of cartilaginous exostoses imaged with MR. PMID- 11405265 TI - Computed tomography diagnosis of isolated splenic torsion in a dog. AB - Isolated splenic torsion in a dog was diagnosed using computed tomography (CT). The enlarged non enhanced spleen and a twisted soft tissue mass effect represented the rotated pedicle were the main findings. Abdominal radiography and ultrasonography were not conclusive. PMID- 11405266 TI - Excretory urography by intraosseous injection of contrast media in a rabbit model. AB - There are many indications for an intravenous excretory urogram. However, where intravenous access is not available, the intraosseous route to the circulation may be an alternative. We found that safe and diagnostic excretory urograms could be obtained in rabbits following the injection of different contrast media via the intraosseous route. In fact, these excretory urograms were indistinguishable from ones obtained by the conventional intravenous route. While the rabbits did not develop any abnormal clinical signs following the procedure, there were postmortem histologic lesions of osteochondrosis in 5 of 22 (22.7%) tibias receiving an intraosseous needle, but in none of the 14 tibias that did not receive an intraosseous needle. Further, the use of diatrizoate was associated with the development of osteochondrosis while the use of iopamidol was not. PMID- 11405267 TI - Radiographic diagnosis: fracture of the caudal aspect of the greater tubercle of the humerus in a horse. PMID- 11405268 TI - Familial cerebellar ataxia with hydrocephalus in bull mastiffs. AB - Familial cerebellar ataxia with concurrent hydrocephalus has previously been described in a family of bull mastiff pups, and recently has been identified in a litter from Louisiana. The 4 affected pups had ataxia, hypermetria, conscious proprioceptive deficits, behavioral abnormalities, and a visual deficit. In magnetic resonance imaging of the brain of two of the pups, there were symmetric hydrocephalus and two focal areas of increased signal intensity within the central nuclei of the cerebellum. Histopathologically there was vacuolization and mild astrogliosis within the deep cerebellar nuclei (dentate, interpositus, fastigial), caudal colliculi, and lateral vestibular nuclei. Although the postmortem results were not exactly the same as in the previously published report, the clinical features and histopathologic findings strongly support the diagnosis. This disorder is most likely inherited in an autosomal recessive manner. PMID- 11405269 TI - Unusual hyperparathyroidism in a cat. AB - A 5 month-old, male, domestic short hair cat was presented with inappetence and vomiting. it was depressed and reluctant to move. The cat had difficulties in keeping the standing position and grossly deformed thighs. Lytic changes and disruption of normal architecture of the bone were observed, involving mainly the femoral diaphyses. An inverse Ca/P ratio and kidney failure were diagnosed. The possibility of whether the bone changes could have been related to primary or secondary renal hyperparathyroidism is discussed. PMID- 11405270 TI - Magnetic resonance imaging of intracranial tissue accumulations in domestic ducks (Anas platyrhynchos f. dom.) with feather crests. AB - The crested breed of domestic duck (Anas platyrhynchos f. dom.) has been described as a variety which has high pre- and postnatal mortalities, malformations in skull and brain anatomy, and several central nervous deficiencies. In addition, intracranial tissue accumulations have been diagnosed in purebred Crested ducks. The incidence, heredity and inheritance of these accumulations as well as their pathogenesis are still generally unknown. The aims of this study were to examine the head of Crested ducks, plain-headed duck breeds, and their crossbreeding relating to the incidence of intracranial alterations. These examinations were performed using magnetic resonance imaging. We found a high incidence of intracranial tissue accumulations in domestic ducks with feather crests. Creasted ducks had more intracranial tissue deposits than plain-headed ducks (p < 0.001). In the present study a correlation between the volume of the crest cushion and the volume of the intracranial tissue deposit could not be found (r = 0.014). Some of the Crested ducks had encephaloceles in addition to the crest cushion. PMID- 11405271 TI - Safety of percutaneous ultrasound-guided biopsy of renal allografts in the cynomolgus monkey: results of 348 consecutive biopsies. AB - The safety of a technique for ultrasound-guided biopsy of renal allografts was evaluated based on 348 consecutive procedures in cynomolgus monkeys. A spring loaded biopsy device with an 18G tru-cut biopsy needle was used to biopsy renal allografts in 139 cynomolgus monkeys performed either on clinical indication (n = 95 animals) or as serial protocol biopsies (n = 44 animals) for a total of 348 biopsies. Monkeys having serial biopsies received between 3-9 biopsies per animal. All others received non-protocol biopsies that were performed on clinical indication, and the range was 1-15 biopsies per animal. No life-threatening complications or deaths occurred and there were no clinically detectable minor complications such as macrohematuria. Self-limiting complications such as small arteriovenous fistulas (n = 4, 3-5 mm large) were detected with Doppler ultrasound and resolved hemodynamically after 2-4 weeks. Three animals developed hematomas ranging 4 mm-2 cm in diameter and were no longer sonographically evident 2-4 weeks later. Ultrasound-guided biopsy of renal allografts can be performed with a high degree of safety in small (3-5 kg) laboratory animals such as the cynomolgus monkey and provides a valuable tool for renal transplantation research. Even when cores were taken at two week intervals no major complications occurred and only rarely were clinically irrelevant complications detected. Experience with diagnostic ultrasound, both gray scale and Doppler, is important for both safety and the recognition of complications that may arise. PMID- 11405272 TI - Echographic examination of the stifle joint affected by cranial cruciate ligament rupture in the dog. AB - Ultrasound is a useful technique for the study of normal and pathologic stifle joints, in particular for soft tissue examination. The aim of this study was to evaluate sonography for examination of stifle joints affected by cranial cruciate ligament rupture. Forty-two medium to giant breed dogs were studied. Tibial compression radiography was performed. A 7.5 MHz transducer with an incorporated 2 cm thick standoff was employed. Sagittal and midsagittal images were collected. The stifle was positioned in maximum flexion during sonography. Sonographic findings were compared with pathologic findings at surgery. Ultrasound was useful in evaluating the presence of fibrous tissue within the joint due to repair processes. It was observed in 70% of stifles with radiographic evidence of chronic osteoarthritis. In 19.6% of the joints it was possible to identify the ruptured cranial cruciate ligament. Ultrasound was not an accurate test for cruciate rupture evaluation, but was specific for the soft tissue pathologic changes which were observed consequent to joint instability. PMID- 11405273 TI - Ultrasonographic imaging of the vagosympathetic trunk in the dog. AB - The purpose of this study was to evaluate the applicability of ultrasonographic imaging of the vagosympathetic trunk in the dog. Cervical ultrasound was performed in 30 healthy dogs. In all 30 dogs the vagosympathetic trunk was detected as a hypoechoic structure in the carotid sheath, adhering the dorsomedial surface of the common carotid artery. The echotexture of the nerve was heterogeneous with anechoic areas separated by hypoechoic bands. A scanner equipped with a 5 to 8 MHz linear array probe was used for imaging and measurements. The diameter of the vagosympathetic trunk ranged from 0.59 to 2.48 mm varying in correlation to the body weight. In summary, ultrasonography is a helpful noninvasive method to image and evaluate the cervical vagosympathetic trunk in the dog. PMID- 11405274 TI - Characterization of coagulase from Staphylococcus intermedius. AB - A protein coagulase was isolated from Staphylococcus intermedius 6131 using bovine prothrombin-Sepharose 4B and Bio-gel P-4 column chromatographies. Homogeneity was demonstrated by the formation of a single band in polyacrylamide gel electrophoresis and isoelectric focusing. The purified preparation possesses a molecular weight of 64,500, an isoelectric point of 4.1, consists of 615 total amino acid residues and demonstrates coagulase activity for human and rabbit fibrinogen, but does not show the activity for rat or guinea pig fibrinogens. This purified protein contains galactose and fucose, and the amino-terminal amino acid sequence was determined. The coagulase activity is inhibited by N bromosuccinimide (NBS), suggesting that tryptophan is involved in this activity. The coagulase was heat stable to 80 degrees C and stable to pH over the range of 7-9. This is the first report of coagulase from Staphylococcus intermedius. PMID- 11405275 TI - Substrate-structure dependence of ribotoxins on cleaving RNA in C. camphora ribosome. AB - Cinnamomin, a type II ribosome-inactivating protein (RIP) isolated from the seeds of Camphora tree (Cinnamomum camphora), could not inactivate its own (autologous) ribosome. Among five RIPs (Cinnamomin A-chain, ricin A-chain, trichosanthin, gelonin, and soporin-S6) tested, only saporin-S6 could cleave the N-glycosidic bond of RNA in C. camphora ribosome to release a specific RNA fragment (R fragment) after treatment with aniline, which was shorter than that from rat liver ribosome. The amount of saporin-S6 to inactivate C. camphora ribosome was about 1000 times higher than that required for rat liver ribosome. Extra ribosomal factors (S-100) in the post-ribosomal supernatant could not promote RNA N-glycosidase activity of cinnamomin and gelonin to C. camphora ribosome. These results indicated that there were some changes in the microenvironments of Sarcin/Ricin domain of C. camphora ribosome that abolished the recognition and catalysis of many RIPs. In addition, the length of C. camphora 5.8S ribosomal RNA was found to be longer than that of rat 5.8S ribosomal RNA. PMID- 11405276 TI - Role of pencycuron in aflatoxin production and cotton seed protection. AB - The research is conducted on the effect of pencycuron and pencycuron-c on cotton seed mycoflora, aflatoxin production and viability. At 8% seed moisture content (mc), pencycuron and pencycuron-c promoted Aspergillus niger, A. flavus, and Penicillium corylophilum growth count at 1 g/kg, but exerted inhibitory effect at 3 and 5 g/kg. At 15% mc, pencycuron enhanced seed-borne fungi at all three doses after most treatment periods (1, 2, 3, 4, and 5 months), whereas pencycuron-c induced inhibition effect. The A. niger utilized pencycuron as nitrogen source more than pencycuron-c. Seeds with 15% mc lost their viability faster than that at 8%, and this was more evident as storage time increased. Such loss occurred faster when seed was treated with pencycuron, whereas pencycuron-c exerted significant activation in the viability compared to the control. The fungal species have high biodegradation activity and produce aflatoxin in different parts of cotton boll (fiber, valves, and seeds). Pencycuron and pencycuron-c inhibited aflatoxin B1 and B2 production in seeds, but did not affect aflatoxin G1 and G2. PMID- 11405277 TI - Prophylactic action of garlic on the histological and histochemical patterns of hepatic and gastric tissues in rats injected with a snake venom. AB - The present study aimed to examine the prophylactic action of oral administration of two doses of garlic on the histological and histochemical patterns of the gastric and hepatic tissues in rats envenomed with cobra snake. The study included the following groups: Group I contained control rats orally administered distilled water for ten days. Group II included rats orally administered daily for ten days with the equivalent therapeutic dose of garlic to rat (18 mg/kg body weight). Group III included rats orally administered daily for ten days with double the equivalent therapeutic dose of garlic to rat (36 mg/kg body weight). Group IV contained rats intramuscularly (i.m.) injected with 1/2 LD50 of cobra venom (0.0125 microg venom/gm body weight) and dissected after 6 hr from injection. Groups V and VI contained rats daily administered with the previous two doses of garlic for ten days, respectively, followed by a single i.m. injection of the above dose of cobra venom after 24 hr from the last garlic application. Rats of these two groups were dissected after 6 hr from venom injection. Administration of the therapeutic dose of garlic induced slight cytoplasmic granulation in some hepatic cells. However, administration of double the therapeutic dose caused swelling, necrosis, and damage of the gastric glandular epithelia together with signs of erosion, exfoliation, and necrosis of the surface mucosal cells. It also induced swelling and coalescence of the hepatic cells, loss of the normal arrangement of the hepatic cords, and hypertrophy of Kupffer cells. Injection with cobra venom caused loss of the normal characteristic appearance of the gastric glands and the epithelial lining cells of the gastric folds and the appearance of numerous inflammatory cells in the lamina properia. It also induced the occurrence of highly swollen hepatic cells, hepatic cellular necrosis and damage, as well as activated Kupffer cells. Nevertheless, pretreatment with the therapeutic dose of garlic for ten days induced a prophylactic activity against the pathogenic effects of the venom in both tissues, which appeared more or less normal except for very minor abnormalities. However, application of double the therapeutic dose of garlic for the same duration did not induce any prophylactic activity. Histochemically, slight alterations were noticed in the polysaccharide, protein, and nucleic acid contents of the gastric mucosa and the hepatic tissues due to administration of the therapeutic doses of garlic. However, severe depletions of these components were recorded in both tissues due to administration of double the therapeutic doses of garlic or injection of cobravenom or the application of both of them together. On the contrary, minor changes were noticed in the histochemical patterns of both tissues in rats pretreated with the therapeutic doses of garlic prior to venom application. It could be concluded that oral administration of the therapeutic dose of garlic for ten days has no serious side effects on gastric and hepatic tissues and could be used as a prophylactic tool against cobra snake envenomation. PMID- 11405278 TI - Analyses of snake venom for forensic investigation. AB - Snake venoms are responsible for two reported human deaths as determined by the double diffusion method. Double diffusion is a simple, rapid, and reliable method, but it is rather qualitative and requires a rather large quantity of sample. In order to evaluate venom concentration more quantitatively, the more sensitive ELISA (enzyme linked immunosorbent assay) method was used as an alternative, and the results were compared with that of the double diffusion method. PMID- 11405279 TI - Toxicity and toxin profiles of the newt, Cynops pyrrhogaster from western Japan. AB - A total of 382 specimens of a Japanese newt, Cynops pyrrhogaster, were collected from western Japan during 1996 to 1999, and assayed for their individual, geographical, sexual, seasonal variations, and anatomical distribution of toxicity by mouse. Most of the specimens tested showed toxicity scores ranging from 5 to 370 MU/g, where no seasonal, but large individual, sexual, and regional variations of toxicity were clearly recognized. Among the parts, skin and muscle showed higher toxicity scores (56 MU/g) than liver, stomach, intestine and gonad, whose toxicity ranged from less than 2 to 33 MU/g. The C. pyrrhogaster toxin was purified by several steps of column chromatography and was shown to consist of tetrodotoxin (TTX) and 6-epiTTX as main components, and 4-epiTTX, 4,9-anhydro-6 epiTTX, and 4,9-anhydroTTX as minor ones by means of HPLC and 1H-NMR analyses. PMID- 11405280 TI - Inhibition of Crotalidae venom hemorrhagic activities by Didelphis marsupialis liver spheroids culture supernatants. AB - The main aim of this work was the development of a primary hepatocyte culture from Didelphis marsupialis, to determine the possible use of culture medium supernatants as a source of inhibitors of the Bothrops lanceolatus venom hemorrhagic activity. The cellular culture was carried out from isolated hepatocytes by the double perfusion technique, and digestion of the liver with collagenase and culturing the hepatocytes in a liquid media under continuous agitation at 37 degrees C in 5% CO2. The hemorrhagic activity inhibition assays were performed inoculating intradermically, a mixture of Bothrops lanceolatus venom plus a pool of liver spheroids culture supernatants, in mice. These liver Didelphis marsupialis spheroid cultures were adequate to obtain large supernatant volumes with inhibitors of hemorrhagic activity. PMID- 11405281 TI - A study on the experimental envenomation in mice with the venom of Tityus trivitattus Kraepelin 1898 (Scorpiones, Buthidae) captured in Argentina. AB - Although Tityus trivitattus is the only scorpion species reported to cause severe human envenomation in Argentina, no previous studies on its venom have been done. Telson homogenates from T. trivitattus specimens collected in Santiago del Estero, Cordoba, and Buenos Aires were employed to study their protein composition and toxicity to mice. Regardless of the site of collection, electrophoretic analysis showed bands at 205, 150, 100, 40, 32, and 13 kDa or smaller. FPLC gel filtration showed three major peaks and 6-8 minor peaks with similar elution volumes. One of the minor peaks from FPLC containing a component of approximately 8 kDa was lethal to mice. Mice injected intravenously with different doses of homogenates presented severe autonomic signs like tachypnea, tachycardia, sialorrhea, lacrimation, profuse sweating, diarrhea, dyspnea, and death. Pathology studies of lungs showed severe congestion of alveolar capillaries, pulmonary edema, and hemorrhagic areas. The kidneys showed glomerular as well as tubular lesions and exocrine glands showed areas of necrosis. The calculated LD50 was 0.38 +/- 0.08 telsons per 20 g mouse, which suggests a lethal potency similar to that of T. serrulatus venom. The lethal potency of 5.0 LD50 of T. trivitattus telson homogenate was neutralized by both an anti-T. trivitattus and a heterologous anti Tityus with ED50 values of 41 +/- 19 and 170 +/- 42 microl, respectively. PMID- 11405282 TI - Mass spectrometry innovations in drug discovery and development. AB - This review highlights the many roles mass spectrometry plays in the discovery and development of new therapeutics by both the pharmaceutical and the biotechnology industries. Innovations in mass spectrometer source design, improvements to mass accuracy, and implementation of computer-controlled automation have accelerated the purification and characterization of compounds derived from combinatorial libraries, as well as the throughput of pharmacokinetics studies. The use of accelerator mass spectrometry, chemical reaction interface-mass spectrometry and continuous flow-isotope ratio mass spectrometry are promising alternatives for conducting mass balance studies in man. To meet the technical challenges of proteomics, discovery groups in biotechnology companies have led the way to development of instruments with greater sensitivity and mass accuracy (e.g., MALDI-TOF, ESI-Q-TOF, Ion Trap), the miniaturization of separation techniques and ion sources (e.g., capillary HPLC and nanospray), and the utilization of bioinformatics. Affinity-based methods coupled to mass spectrometry are allowing rapid and selective identification of both synthetic and biological molecules. With decreasing instrument cost and size and increasing reliability, mass spectrometers are penetrating both the manufacturing and the quality control arenas. The next generation of technologies to simplify the investigation of the complex fate of novel pharmaceutical entities in vitro and in vivo will be chip-based approaches coupled with mass spectrometry. PMID- 11405283 TI - A novel drug-regulated gene expression system based on the nuclear receptor constitutive androstane receptor (CAR). AB - PURPOSE: To develop and characterize a new drug-regulated gene expression system based on the nuclear receptor constitutive androstane receptor (CAR). METHODS: Both transient and stable transfection into HEK293 cells of luciferase plasmids under the control of either drug- and steroid-responsive nuclear receptor CAR or the tetracycline-sensitive transactivator tTA were used in development of stable cell lines. RESULTS: A stable first-generation cell line that expresses luciferase gene under the control of nuclear receptor CAR was developed. The luciferase expression in CAR-producing cells could be suppressed by androstanes and reactivated by structurally unrelated drugs chlorpromazine, metyrapone, phenobarbital, and clotrimazole. The kinetics of luciferase expression in CAR producing cells and the tTA system were comparable. The overall regulation of CAR system was improved by modifications to the DNA binding domain and site. CONCLUSIONS: Because of its wide ligand selectivity and transferable ligand binding domain, CAR expands the repertoire of regulated gene expression systems. PMID- 11405284 TI - Flurbiprofen enantiomers inhibit inducible nitric oxide synthase expression in RAW 264.7 macrophages. AB - PURPOSE: Using RAW 264.7 macrophages, the present study investigates the influence of optically pure enantiomers of the nonsteroidal anti-inflammatory drug flurbiprofen on lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) expression. METHODS: iNOS and cyclooxygenase-2 (COX-2) mRNA levels were measured by quantitative real-time reverse-transcription polymerase chain reaction (RT-PCR). Concentrations of nitrite (index of cellular NO production) and prostaglandin E2 (index of COX-2 activity) in cell culture supernatants were determined by Griess assay and enzyme immunoassay, respectively. RESULTS: R(-)- and S(+)-flurbiprofen decreased LPS-induced iNOS mRNA and nitrite levels in an equipotent and concentration-dependent manner. Suppression of iNOS mRNA expression by R(-)- and S(+)-flurbiprofen was gene specific in that both substances failed to inhibit LPS-induced COX-2 mRNA expression. By contrast, flurbiprofen enantiomers suppressed LPS-induced prostaglandin E2 formation enantioselectively with S(+)-flurbiprofen being considerably more potent than its R(-)-antipode. CONCLUSIONS: Our results show that R(-)- and S(+)-flurbiprofen, albeit differing in their potency as inhibitors of COX-2 activity, equipotently suppress iNOS expression. Because sustained high NO levels are associated with pain and tissue injury under various pathological conditions, a suppression of the inducible NO pathway may contribute to the pharmacological action of both R(-)- and S(+)-flurbiprofen. PMID- 11405285 TI - Studies on agents with mixed NO-dependent and calcium channel antagonistic vasodilating activities. AB - PURPOSE: To obtain new cardiovascular agents with mixed Ca2+-channel antagonistic and NO-donor properties, a series of "hybrid" 1,4-dihydropyridines (1,4-DHPs), bearing NO-donating furoxan moieties on the 3-positioned lateral ester chain were synthesized and pharmacologically characterized. Furazan analogues were also prepared and investigated for control purposes, because they are unable to release NO. METHODS: Synthesis of the models was achieved by a modified Hantzsch approach. All of the final furoxan 1,4-DHPs were assessed for their ability to produce nitrite in the presence of a large excess of cysteine by the Griess reaction. Vasodilating activity was evaluated on rat aorta and expressed as EC50 and EC50MB values, obtained in the absence and in the presence of methylene blue (MB) respectively, a well-known guanylate cyclase inhibitor. Affinities to 1,4 DHP receptor on Ca2+-channels, expressed as IC50 values, were determined through displacement experiments of [3H]-nitrendipine on rat cortex homogenates. RESULTS: Some hybrid compounds (derivatives 15a, 15b, 16a, and 16b) displayed vasodilating activity depending predominantly on their Ca2+-channel blocker properties. By contrast, some others (derivatives 17a, 17b, and 21) behaved as well-balanced hybrids with mixed Ca2+-channel blocking and NO-dependent vasodilating activities. CONCLUSION: This work demonstrates the possibility of obtaining well balanced hybrids endowed with mixed NO-donor and Ca2+-channel blocker properties using appropriate 1,4-DHP and furoxan moieties. A procedure for the individual evaluation of the NO-dependent vasodilator component and that due to Ca2+-channel blocking is proposed. PMID- 11405286 TI - Risedronate pharmacokinetics and intra- and inter-subject variability upon single dose intravenous and oral administration. AB - PURPOSE: To determine the pharmacokinetics and absolute bioavailability of risedronate after single-dose oral administration of 30 mg risedronate as a tablet and an aqueous solution, and 0.3 mg risedronate as an intravenous infusion. METHODS: This study was a randomized, three-treatment, four-period, partial replicate crossover study involving 33 healthy volunteers. Treatments were administered 7 weeks apart, and the third treatment was repeated during the fourth period. Serum and urine were collected over 72 hours and 672 hours, respectively. RESULTS: Following intravenous administration, renal clearance accounted for 87% of total clearance, with 65% of the dose excreted within 24 hours and 85% of the dose excreted within four weeks. The absolute bioavailability was approximately 0.62% after both oral formulations, and the relative bioavailability of the tablet compared with the oral solution was 104%. The rate and extent of absorption from the two formulations were bioequivalent based on the range proposed for highly variable drugs. Intrasubject variability following oral administration was 50-80%, and was primarily associated with absorption. CONCLUSION: The majority of the total clearance after intravenous administration of risedronate was renal clearance, indicating that only a small percentage of a systemic dose is potentially incorporated, or "cleared," into bone. The absolute bioavailability of orally administered risedronate is approximately 0.6%, and is independent of formulation. Variability in the pharmacokinetics following oral administration is primarily associated with intrasubject variability in absorption. PMID- 11405287 TI - A functional assay for quantitation of the apparent affinities of ligands of P glycoprotein in Caco-2 cells. AB - PURPOSE: To develop a facile functional assay for quantitative determination of the apparent affinities of compounds that interact with the taxol binding site of P-glcoprotein (P-gp) in Caco-2 cell monolayers. METHODS: A transport inhibition approach was taken to determine the inhibitory effects of compounds on the active transport of [3H]-taxol, a known substrate of P-gp. The apparent affinities (K(I) values) of the compounds were quantitatively determined based on the inhibitory effects of the compounds on the active transport of [3H]-taxol. Intact Caco-2 cell monolayers were utilized for transport inhibition studies. Samples were analyzed by liquid scintillation counting. RESULTS: [3H]-Taxol (0.04 microM) showed polarized transport with the basolateral (BL) to apical (AP) flux rate being about 10-20 times faster than the flux rate in the AP-to-BL direction. This difference in [3H]-taxol flux could be totally abolished by inclusion of (+/-) verapamil (0.2 mM), a known inhibitor of P-gp, in the incubation medium. However, inclusion of probenecid (1.0 mM), a known inhibitor for the multidrug resistance associated protein (MRP), did not significantly affect the transport of [3H] taxol under the same conditions. These results suggest that P-gp, not MRP, was involved in taxol transport. Quinidine, daunorubicin, verapamil, taxol, doxorubicin, vinblastine, etoposide, and celiprolol were examined as inhibitors of the BL-to-AP transport of [3H]-taxol with resulting K(I) values of 1.5+/-0.8, 2.5+/-1.0, 3.0+/-0.3, 7.3+/-0.7, 8.5+/-2.8, 36.5+/-1.5, 276+/-69, and 313+/-112 microM, respectively. With the exception of that of quinidine, these K(I) values were comparable with literature values. CONCLUSIONS: This assay allows a facile quantitation of the apparent affinities of compounds to the taxol-binding site in P-gp, however, this assay does not permit the differentiation of substrates and inhibitors. The potential of drug-drug interactions involving the taxol binding site of P-gp can be conveniently estimated using the protocol described in this paper. PMID- 11405288 TI - Effects of furanocoumarin derivatives in grapefruit juice on nifedipine pharmacokinetics in rats. AB - PURPOSE: It has been reported that grapefruit juice (GJ) causes a pharmacokinetic interaction with many drugs after co-ingestion. It is postulated that the substances in GJ may inhibit the first-pass metabolism during the intestinal absorption process. In recent years, several furanocoumarin derivatives that inhibit P450 activity in intestinal microsomes were isolated from GJ. In this study, we report the effects of the furanocoumarin derivatives in GJ on the nifedipine (NFP) pharmacokinetics in rats. METHODS: Three furanocoumarin derivatives (bergaptol [BT], bergamottin [BG], and 6',7'-dihydroxybergamottin [DHB]) found in GJ were used in this study. Each furanocoumarin was reconstituted in orange juice at the same concentration as in the GJ. Two milliliters of each sample was administered into the rat duodenum. After 30 min, NFP was intraduodenally administered at a dose of 3 mg/kg body weight. The NFP concentrations in the plasma samples were determined by HPLC. RESULTS: A significant increase in the AUC of NFP was observed only in the rats administered BG; 1.5 times that of the control group. The result was quite identical with that of the group that was administered GJ. BT and DHB had no significant effects on the NFP pharmacokinetics. CONCLUSIONS: The results strongly suggested that BG in GJ might be the substance that elevates the NFP plasma concentrations. PMID- 11405289 TI - Screening of multidrug-resistance sensitive drugs by in situ brain perfusion in P glycoprotein-deficient mice. AB - PURPOSE: This study was conducted to assess the influence of P-glycoprotein (P gp) on brain uptake of multidrug resistance sensitive drugs using an in situ brain perfusion technique in P-gp-deficient (mdr1a[-/-]) and wild-type mice. METHODS: The blood-brain transport of radiolabeled vinblastine, vincristine, doxorubicin, colchicine, and morphine was evaluated in mdr1a(-/-) and wild-type CF-1 mice with the in situ brain perfusion technique. Brain uptake of drugs after intravenous pretreatment with P-gp reversal agents, (PSC 833, GF 120918, or (+/-) verapamil), or vehicle also was studied in wild-type mice. In all experiments, cerebral vascular volume was determined by co-perfusion of sucrose. RESULTS: Cerebral vascular volume was preserved during perfusion, indicating maintenance of blood-brain barrier integrity in both types of mice within the concentration range of substrates in the perfusate. The apparent brain transport of colchicine. vinblastine, doxorubicin, and morphine was increased 3.0, 2.7, 1.5, and 1.4-fold, respectively, in mdr1a(-/-) mice compared with the wild-type: the brain uptake of vincristine was not affected by P-gp. Preadministration of PSC 833 or GF 120918 in wild-type mice led to a -3-fold increase in the brain transport of colchicine and vinblastine, but no effect was observed for the other compounds. Intravenous verapamil enhanced colchicine brain transport (1.8-fold), but failed to increase the brain uptake of vinblastine and morphine. CONCLUSION: The in situ brain perfusion technique appears to be a sensitive and powerful tool for medium throughput screening of the brain uptake of multidrug resistance sensitive drugs. The effect of P-gp is characterized more efficiently with mdr1a(-/-) mice than by using modulators of P-gp in wild-type mice. PMID- 11405290 TI - Tyrphostin-8 enhances transferrin receptor-mediated transcytosis in Caco-2- cells and inreases hypoglycemic effect of orally administered insulin-transferrin conjugate in diabetic rats. AB - PURPOSE: To investigate the effect of tyrphostin 8 (T-8), a GTPase inhibitor, on transferrin receptor (TfR)-mediated transcytosis of insulin-transferrin (In-Tf) conjugate in cultured enterocyte-like Caco-2 cells and on gastrointestinal (GI) absorption of In-Tf in streptozotocin (STZ)-induced diabetic rats. METHODS: Caco 2 cells and diabetic rats were used as in vitro and in vivo models, respectively. TfR-mediated transcytosis was measured using 125I-In-Tf. The absorption of insulin in diabetic rats was demonstrated by the hypoglycemic effect. Rat blood glucose level was determined using a ONE TOUCH blood glucose monitoring system. RESULTS: T-8 increased apical-to-basolateral transport of In-Tf conjugate by enhancing TfR-mediated transcytosis in filter-grown Caco-2 cell monolayer, and this enhancement was higher and faster than the previously reported brefeldin A (BFA)-induced effect. The measurement of transepithelial electrical resistance (TEER) during the transport study showed that T-8 was less destructive on the cell tight junction than BFA. The GI absorption of In-Tf was evaluated by its hypoglycemic effect after oral administration in STZ-induced diabetic rats. The glucose-lowering effect of orally administered In-Tf in STZ-induced diabetic rats was improved by either T-8 or BFA. However, the effect of T-8 was more potent than that of BFA, especially at 7 h after administration. Either non-conjugated insulin or insulin-human serum albumin (In-HSA) conjugate by itself or in combination with T-8 did not show any hypoglycemic effect after oral administration, indicating that T-8-enhanced hypoglycemic activity of In-Tf was due to a selective enhancement of TfR-mediated transcytosis. CONCLUSIONS: Our data indicated that T-8 could be used to increase the GI absorption of insulin as a transferrin conjugate. T-8, as an enhancer of TfR-mediated transcytosis, is better than the previously reported BFA. T-8 produces a higher increase on the transport of In-Tf and a lower toxicity on epithelial cells. Our findings provide an alternative approach to promote the GI absorption of insulin, as well as other peptide or protein drugs. PMID- 11405291 TI - Characterization of murine monoclonal antibody to tumor necrosis factor (TNF-MAb) formulation for freeze-drying cycle development. AB - PURPOSE: This study was designed to characterize the formulation of protein pharmaceuticals for freeze-drying cycle development. Thermal properties of a protein formulation in a freezing temperature range are important in the development of freezing and primary drying phases. Moisture sorption properties and the relationship between moisture and stability are the bases for the design of the secondary drying phase. METHODS: We have characterized the formulation of TNF-MAb for the purpose of freeze-drying cycle development. The methods include: DTA with ER probes, freeze-drying microscopy, isothermal water adsorption, and moisture optimization. RESULTS: The DTA/ER work demonstrated the tendency to "noneutectic" freezing for the TNF-MAb formulation at cooling rates of -1 to -3 degrees C/min. The probability of glycine crystallization during freezing was quite low. A special treatment, either a high subzero temperature holding or annealing could promote the maximum crystallization of glycine, which could dramatically increase the Tg' of the remaining solution. The freeze-drying microscopy further indicated that, after the product was annealed, the cake structure was fully maintained at a Tp below -25 degrees C during primary drying. The moisture optimization study demonstrated that a drier TNF-MAb product had better stability. CONCLUSIONS: An annealing treatment should be implemented in the freezing phase in order for TNF-MAb to be dried at a higher product temperature during primary drying. A secondary drying phase at an elevated temperature was necessary in order to achieve optimum moisture content in the final product. PMID- 11405292 TI - Assessing tablet bond types from structural features that affect tablet tensile strength. AB - PURPOSE: The aim of this article was to study the possibility of assessing the structural features affecting tablet tensile strength to obtain information on the dominating bond types, i.e. interparticulate attractions, in tablets. METHODS: The features of the internal tablet structure considered to be important for tablet tensile strength were assessed using a simple tablet model for tablets made from seven materials: potassium chloride, sodium chloride, sodium bicarbonate, lactose, sucrose, microcrystalline cellulose, and ascorbic acid. RESULTS: Tablet porosity and particle size (measured as external specific surface area by permeametry) were the structural features that best correlated with tablet tensile strength. These features were described by a "structural factor," which was combined with tablet tensile strength, as an "interaction factor," to reflect the dominating bond types in tablets. CONCLUSION: The qualitative results gave dominating bond types in the tablets studied that matched the results of earlier studies, thus supporting the applicability of the method. PMID- 11405293 TI - Thiolated polymers: development and evaluation of transdermal delivery systems for progesterone. AB - PURPOSE: To evaluate the possible use of polycarbophil-cysteine (PCP-Cys) as polymeric matrix for transdermal progesterone application. METHODS: Thiolated polycarbophil was synthesised by the covalent attachment of cysteine to the basis polymer. The adhesive properties of PCP-Cys in comparison to polyvinylpyrrolidone/hydroxypropylmethylcellulose (PVP/HPMC) and polyvinylpyrrolidone/polyvinylalcohol (PVP/PVA) were investigated by testing the total work of adhesion (TWA) on porcine skin. Release studies in Franz diffusion cells and standard in vitro permeation experiments with porcine skin were performed analysing the progesterone content by high-performance liquid chromatography. RESULTS: Films based on PCP-Cys displayed very high cohesive properties due to the formation of interchain disulfide bonds. The TWA of the thiolated polymer on porcine skin was significantly (P <0.05) the highest. In addition progesterone permeation was also the highest from PCP-Cys compared with PVP/HPMC and PVP/PVA within 24 hours. CONCLUSION: PCP-Cys--a partly thiolated polymer--might be a novel polymer matrix for transdermal progesterone delivery with excellent adhesiveness on porcine skin. PMID- 11405294 TI - Evaluation of carboxymethylpullulan as a novel carrier for targeting immune tissues. AB - PURPOSE: To demonstrate the potential of carboxymethylpullulan (CMPul) as a carrier for targeting immune tissues, and to find whether immune tissues could be set as the target of an immunosuppressant to treat autoimmune diseases. METHODS: The biodistribution of CMPul was investigated to evaluate its potency as a carrier for targeting immune tissues. Furthermore, an immunosuppressant-CMPul conjugate was prepared and its suppressive effect on rat adjuvant arthritis was examined. RESULTS: The disappearance rate of 3H-labeled CMPul from the blood circulation was much slower than that of 3H-labeled pullulan (Pul) after intravenous injection to normal rats. The concentration of 3H-labeled CMPul in the spleen and lymph nodes was much higher than that of 3H-labeled Pul at 24 hours after the injection, whereas the concentration of 3H-labeled CMPul in the liver was significantly lower than that of 3H-labeled Pul. A similar targeting property of 3H-labeled CMPul for these immune tissues was observed in arthritic rats. A conjugate composed of a novel immunosuppressant PA-48153C and CMPul showed a suppressive effect on rat adjuvant arthritis judging from a reduction of the arthritic index and spleen weight and an increase of body weight. CONCLUSIONS: CMPul is expected to be a promising carrier for targeting immune tissues with an immunosuppressant to enable treatment of autoimmune diseases. PMID- 11405295 TI - Use of labor economic theory to examine hours worked by male and female pharmacists. AB - PURPOSE: The objectives of this study were to develop a theoretically derived model of hours worked by pharmacists and estimate the model separately for male and female pharmacists. METHODS: A systematic random sample of 1,600 pharmacists from four states was mailed a survey asking about current and past employment information. Two dependent variables were studied: weekly hours worked and annual hours worked. Independent variables were categorized as economic variables (hourly wage rate, other income, total debt) and demographic variables (employment position, age, degree earned, marital status, number of children at home). A two equation multiple regression model was estimated with two-stage least squares regression. RESULTS: A total of 541 pharmacists responded to the survey and data from 442 of the respondents were used in the analysis. Hourly wage rates were negatively associated with weekly hours worked for males. Other income and total debt were significantly negatively and positively associated, respectively, with annual hours worked by female pharmacists. The number of young children at home significantly reduced weekly and annual hours worked by female pharmacists. Female pharmacists earning a Pharm.D. degree worked significantly more hours weekly and annually. Age was significantly negatively associated with male pharmacists weekly and annual hours worked. CONCLUSIONS: Economic variables had a relatively small effect on hours worked by male and female pharmacists suggesting that increased wage rates may not increase hours worked. Strategies to increase hours worked by females likely should focus on benefits to help females handle childcare issues. PMID- 11405296 TI - Degradation kinetics of oxycarbonyloxymethyl prodrugs of phosphonates in solution. PMID- 11405297 TI - Relationship of pharmacokinetically-calculated volumes of distribution to the physiologic distribution of liposomal drugs in tissues: implications for the characterization of liposomal formulations. PMID- 11405298 TI - Synergistic effect of indomethacin and bleomycin on tumor growth produced by activating antitumor immunity. PMID- 11405299 TI - Nocodazole treatment of CV-1 cells enhances nuclear/perinuclear accumulation of lipid-DNA complexes and increases gene expression. PMID- 11405300 TI - On the electrical properties of dislocations in ZnS using electric force microscopy. AB - A combination of electric force microscopy (EFM) and noncontact atomic force microscopy (AFM) was used to study microscratching-induced dislocations in sphaleritic ZnS single crystals. Dislocation bands predominantly consisting of either anion-type (S) or cation-type (Zn) dislocations were induced by scratching along either [111] or [111] on a (110) surface. A significant difference of local distortions in electrical potential between the S(g) and Zn(g) dislocation bands was observed from the EFM images. Electric charges of these dislocations were determined quantitatively and the results were compared with theoretical models. PMID- 11405301 TI - Image analysis of particle dispersions in microscopy images of cryo-sectioned sausages. AB - Two feature extraction methods, the three-dimensional (3-D) local box-counting method and the area distribution method, are presented to describe the fat dispersion pattern on digital microscopy images of cryo-sectioned sausages. Both methods calculate whole arrays of variables for each microscopy image. The 3-D box-counting method calculates scale dependent (local) dimensions. This is in contrast to common fractal methods, which are univariate. Principal component analysis (PCA) was used to show that different sausages yield different fat dispersion patterns. Partial least square regression (PLS) shows that there is a correlation between the variables gained with both methods and the fat content. PMID- 11405302 TI - Improvement of DNA-visualization in dynamic mode atomic force microscopy in air. AB - Near-contact mode atomic force microscopy (AFM) imaging leads to sharper representations of DNA double strands on mica imaged at ambient conditions compared with noncontact mode AFM. Phase shift was used for feedback control yielding height information using a simple model calculation. No contact between tip and sample occurs. Measured DNA widths were up to four times smaller than measured with the same AFM tip in noncontact mode at ambient condition. PMID- 11405303 TI - Mapping piezoelectric-field distribution in gallium nitride with scanning second harmonic generation microscopy. AB - Taking advantage of the electric field-enhanced second-harmonic generation effect in bulk gallium nitride (GaN) and indium gallium nitride (InGaN) quantum wells, we demonstrated the piezoelectric field distribution mapping in bulk GaN and InGaN multiple-quantum-well (MQW) samples using scanning second-harmonic generation (SHG) microscopy. Scanning SHG microscopy and the accompanying third harmonic generation (THG) microscopy of the bulk GaN sample were demonstrated using a femtosecond Cr:forsterite laser at a wavelength of 1230 nm. Taking advantage of the off-resonant electric field-enhanced SHG effect and the bandtail state-resonance THG effect, the second- and third-harmonic generation microscopic images obtained revealed the piezoelectric field and bandtail state distributions in a GaN sample. Combined with 720 nm wavelength excited two-photon fluorescence microscopy in the same sample, the increased defect density around the defect area was found to suppress bandedge photoluminescence, to increase yellow luminescence, to increase bandtail state density, and to decrease residue piezoelectric field intensity. Scanning SHG microscopy of the InGaN MQW sample was resonant excited with 800 nm femtosecond pulses from a Ti:sapphire laser in order to suppress SHG contribution from the bulk GaN substrate. Taking advantage of the strong piezoelectric field inside the InGaN quantum well, the wavelength resonant effect, and the electric field-enhanced SHG effect of InGaN quantum wells, resonant scanning SHG microscopy revealed the piezoelectric field distribution inside the wells. Combined with accompanying three-photon fluorescence microscopy from the bulk GaN substrate underneath the quantum wells, the direct correspondence between the piezoelectric field strength inside the quantum well and the substrate quality can be obtained. According to our study, the GaN substrate area with bright bandedge luminescence corresponds to the area with strong SHG signals indicating a higher stained-induced piezoelectric field. These scanning harmonic generation microscopies exhibit superior images of the piezoelectric field and defect state distributions in GaN and InGaN MQWs not available before. Combining with scanning multiphoton fluorescence microscopy, these techniques open new ways for the physical property study of this important material system and can provide interesting details that are not readily available by other microscopic techniques. PMID- 11405304 TI - FERImage: an interactive program for fractal dimension, d(per) and d(min) calculation. AB - A computer program has been written for the determination of the D fractal dimension at low scale, of the d(per) representative parameter of the periodical region at high scale, and the d(min), representative parameter of the minimum elemental cell which is repeated in the periodical structure from the variogram. It carries out the simultaneous obtention of the three previous parameters developed by Bonetto and Ladaga. The program also allows to obtain fractal dimension values from the Fourier power spectrum. FERImage has been developed so that the users could choose the rank where the behavior is fractal, not only in the variogram method but also in the Fourier spectrum method. PMID- 11405305 TI - Investigation of corrosion and container integrity in metal food containers using scanning electron microscopy-X-ray microanalysis. AB - Scanning electron microscopy and x-ray microanalysis were used to investigate microleakage, two incidences of loss of coating integrity, filiform corrosion, and sulfide black corrosion in enamel-lined metal food containers. Microleakage developed in a canned cheese sauce at the side seam weld and was traced to a hot weld and a cold weld defect. An enamel-blistering problem developed in an aseptically processed milk-based product when the tinplate was heated above its melting point; the tin melted under the enamel coating, causing the blistering. An enamel-blistering problem, due to silicon contamination of the metal surface prior to enameling, developed on the bottom of a two-piece can that contained a chicken-based product. A sulfide black problem developed in canned clams, caused by scratches in the coating that were introduced during can manufacturing. A filiform corrosion problem developed on the outside of the easy score line of enameled tinplate lids with a light tin coating 0.08 microns thick. PMID- 11405306 TI - A high-resolution add-on in-lens attachment for scanning electron microscopes. AB - A compact add-on objective lens for the scanning electron microscope (SEM) has been designed and tested. The lens is < 35 mm high and can be fitted on to the specimen stage as an easy-to-use attachment. Initial results show that it typically improves the spatial resolution of the SEM by a factor of three. The add-on unit is based upon a permanent magnet immersion lens design. Apart from the extra attachment to the specimen stage, the SEM with the add-on lens functions in the normal way. The in-lens unit can comfortably accommodate specimen heights up to 10 mm. The new add-on lens unit opens up the possibility of operating existing SEMs in the high-resolution in-lens mode. By using a deflector at the top of the add-on lens unit, it can also operate as a quantitative multichannel voltage contrast spectrometer, capable of recording the energy spectrum of the emitted secondary electrons. Initial experiments confirm that a significant amount of voltage contrast can be obtained. PMID- 11405307 TI - A longitudinal analysis of anxiety and depressive symptoms. AB - The authors modeled depressive and anxiety symptom data from 1,391 participants in a longitudinal study of middle-aged and older Swedish twins (M age = 60.9 years, SD = 13.3). Although anxiety and depression were highly correlated, a model with distinct Anxiety and Depression factors fit the data better than models with Positive and Negative Affect factors or a single Mental Health factor. Lack of well-being was associated with anxiety rather than depression. Over two 3-year intervals, anxiety symptoms led to depressive symptoms, but the relationship was not reciprocal. Anxiety symptoms were more stable than depression. These findings provide additional support for the idea that anxiety symptoms may reflect a personality trait such as neuroticism more than do depressive symptoms and suggest that low positive affect may not be as specific to depression among older adults as in younger people. PMID- 11405308 TI - The strong connection between sensory and cognitive performance in old age: not due to sensory acuity reductions operating during cognitive assessment. AB - Cognitive aging research has documented a strong increase in the covariation between sensory and cognitive functioning with advancing age. In part, this finding may reflect sensory acuity reductions operating during cognitive assessment. To examine this possibility, the authors administered cognitive tasks used in prior studies (e.g., Lindenberger & Baltes, 1994) to middle-aged adults under age-simulation conditions of reduced visual acuity, auditory acuity, or both. Visual acuity was lowered through partial occlusion filters, and auditory acuity through headphone-shaped noise protectors. Acuity manipulations reduced visual acuity and auditory acuity in the speech range to values reaching or approximating old-age acuity levels, respectively, but did not lower cognitive performance relative to control conditions. Results speak against assessment related sensory acuity accounts of the age-related increase in the connection between sensory and cognitive functioning and underscore the need to explore alternative explanations, including a focus on general aspects of brain aging. PMID- 11405309 TI - Planning for the future: a life management strategy for increasing control and life satisfaction in adulthood. AB - The study examined the social, personality, and cognitive correlates of self reported future planning and the relationship of future planning to perceived control and life satisfaction. Using 2 probability samples of adults ages 25-74 (n = 2,971, n2 = 300) findings suggest, for Study 1, that education, income, social support, predictability, Conscientiousness. and Openness to Experience were positively related to future planning, whereas Neuroticism and Agreeableness were negatively related. Men were more future oriented; as age increased, future planning decreased. Study 2 replicated the findings with the exception of age, Agreeableness, and Neuroticism. For both studies, results supported a model in which the effects of future planning on life satisfaction were mediated by sense of control. A Planning x Age interaction for Study I indicated that although self reported future planning decreased with age, the positive effects of future oriented planning strategies on life satisfaction were most pronounced for the older adults, and this relationship was also mediated by control beliefs. PMID- 11405310 TI - Relationship quality and potentially harmful behaviors by spousal caregivers: how we were then, how we are now. The Family Relationships in Late Life Project. AB - Structured interview data from 142 caregivers (98 wives, 44 husbands) indicate that more depressed caregivers are more likely to treat their spouses in potentially harmful ways. However, consistent with hypotheses derived from communal relationships theory, when the preillness relationship between caregiver and care recipient was characterized by mutual responsiveness to each other's needs (i.e., was more communal), caregivers were less depressed and less frequently engaged in potentially harmful behaviors. These effects were not attributable to demographic factors, amount of care provided, care recipient dementia status, or length of time in the caregiving role. Rather, multivariate analyses suggest that the extent to which premorbid relationships were communal in nature determines whether caregivers perceive their current relationships as rewarding, which, in turn, predicts caregiver depression and potentially harmful behaviors. PMID- 11405311 TI - Language decline across the life span: findings from the Nun Study. AB - The present study examines language samples from the Nun Study. Measures of grammatical complexity and idea density were obtained from autobiographies written over a 60-year span. Participants who had met criteria for dementia were contrasted with those who did not. Grammatical complexity initially averaged 4.78 (on a 0-to-7-point scale) for participants who did not meet criteria for dementia and declined .04 units per year; grammatical complexity for participants who met criteria for dementia initially averaged 3.86 and declined .03 units per year. Idea density averaged 5.35 propositions per 10 words initially for participants who did not meet criteria for dementia and declined an average of .03 units per year, whereas idea density averaged 4.34 propositions per 10 words initially for participants who met criteria for dementia and declined .02 units per year. Adult experiences, in general, did not moderate these declines. PMID- 11405312 TI - Effects of goal-setting and feedback on memory performance and beliefs among older and younger adults. AB - This research examined the impact of goal-setting conditions on memory beliefs and performance among older and younger adults. After baseline recall and assessment of beliefs, participants were assigned to goal-setting, goals plus feedback, or control. Then, additional recall trials were followed by repeated memory beliefs assessments. For both younger and older adults, performance, motivation, and self-efficacy were affected positively by goal-setting. The impact of goals plus feedback was mixed and varied as a function of age and dependent measure. Success rates for reaching memory goals, which were low for the older adults, may have been a factor in these results. Adults' self-set recall goals were predicted initially by baseline performance and self-efficacy. On the final trial, goals were predicted by last trial performance, self efficacy, and control beliefs. PMID- 11405313 TI - Attempted decomposition of age-related influences on two tests of reasoning. AB - Two studies were designed to determine which of several hypothesized components were primarily responsible for the age-related effects on matrix reasoning (Study 1) and analytical reasoning (Study 2). In neither case were the analyses successful because the variables selected to assess the hypothesized components failed to exhibit discriminant validity and also had little unique age-related influences. These results. along with the results of other analyses, suggest that large proportions of the age-related effects on different cognitive variables are shared and are not independent of one another. Implications of these findings for the interpretation of age-related influences on measures of cognitive functioning are discussed. PMID- 11405314 TI - Primary versus secondary insomnia in older adults: subjective sleep and daytime functioning. AB - Most psychological research on insomnia has centered on primary insomnia (PI). Secondary insomnia (SI), though more common than PI, has received little attention because of its presumed unresponsiveness to treatment. The present study recruited older adults with PI, SI, and a comparison group of older adults with no insomnia (NI). Self-report assessments of sleep revealed no significant difference between the 2 insomnia groups. Daytime functioning measures found significant differences in impairment between the 3 groups with SI having the worst daytime functioning, followed by PI, which was worse than NI. Further analyses found substantial independence between sleep and daytime functioning. Implications of these findings for the clinical management of SI are discussed. PMID- 11405315 TI - The heritability of cognitive functioning in very old adults: evidence from Danish twins aged 75 years and older. AB - Heritable influences on cognitive functioning were investigated in a sample of 403 pairs of like-sex Danish twins aged 75 years and older. Twins completed the Mini-Mental State Examination and 3 other cognitive tests. Genetic factors accounted for 26-54% of the variance on these measures, with the balance being due to environmental factors that create differences rather than similarities among reared-together relatives. Deleting twins with severe cognitive impairment had little effect on the results, indicating that the heritability of cognitive functioning was not due entirely to genes affecting dementia. Neither age nor gender moderated twin similarity, and differential social contact could not account for correlation differences between monozygotic and dizygotic twins. These results replicate G. E. McClearn et al.'s (1997) study in indicating substantial genetic influences on late-life cognitive functioning. PMID- 11405316 TI - Binocular detection of masked patterns in young and old observers. AB - A visual pattern embedded in noise is detected appreciably better when the stimulus complex contains interocular cues (dichoptic condition) than when such cues are absent (binoptic condition). In a recent study (F. Speranza, G. Moraglia, & B. A. Schneider, 1995) the authors showed that the relative difference between binoptic and dichoptic thresholds does not change with age. However, older adults showed higher binoptic and dichoptic thresholds, thus suggesting an age-related difficulty with degraded stimulation. In this article the authors first replicated these findings and proceeded next to investigating whether age-related changes in processing efficiency, additive internal noise, and the spatial frequency bandwidth of the detecting filters could account, separately or concurrently, for the elevated thresholds in noise exhibited by the older adults. Results indicate that this increase is not attributable to age related changes in filter bandwidth or internal noise. Rather, the findings can be explained in terms of a decrease in processing efficiency with age. PMID- 11405317 TI - Expertise and age-related changes in components of intelligence. AB - In a sample of 263 male GO players at 48 levels of expertise and ranging from 18 to 78 years of age, it was found that factors of expertise deductive reasoning (EDR) and expertise working memory (EWM) were independent of factors of fluid reasoning (Gf) and short-term working memory (STWM) that, along with cognitive speed (Gs), have been found to characterize decline of intelligence in adulthood. The main effects of analyses of cross-sectional age differences indicated age related decline in EDR and EWM as well as in Gf, STWM, and Gs. However, interaction and partialing analyses indicated that decline in EDR and EWM decreased to no decline with increase in level of expertise. The results thus suggest that with increase in factors known to raise the level of expertise- particularly, intensive, well-designed practice--there may be no age-related decline in the intelligence that is measured in the abilities of expertise. PMID- 11405318 TI - The structure of verbal abilities in young and older adults. AB - Four language sample measures as well as measures of vocabulary, verbal fluency, and memory span were obtained from a sample of young adults and a sample of older adults. Factor analysis was used to analyze the structure of the vocabulary, fluency, and span measures for each age group. Then an "extension" analysis was performed by using structural modeling techniques to determine how the language sample measures were related to the other measures. The measure of grammatical complexity was associated with measures of working memory including reading span and digit span. Two measures, sentence length in words and a measure of lexical diversity, were associated with the vocabulary measures. The fourth measure, propositional density, was associated with the fluency measures as a measure of processing efficiency. The structure of verbal abilities in young and older adults is somewhat different, suggesting age differences in processing efficiency. PMID- 11405319 TI - The effects of aging on reaction time in a signal detection task. AB - The effects of aging on response time are examined in 2 simple signal detection tasks with young and older subjects (age 60 years and older). Older subjects were generally slower than young subjects, and standard Brinley plot analyses of response times showed typical results: slopes greater than 1 and (mostly) negative intercepts. R. Ratcliff, D. Spieler, and G. McKoon (2000) showed that the slopes of Brinley plots measure the relative standard deviations of the distributions of response times for older versus young subjects. Applying R. Ratcliff's (1978) diffusion model to fit the response times, their distributions, and response accuracy, it was found that the larger spread in older subjects' response times and their slowness relative to young subjects comes from a 50-ms slowing of the nondecision components of response time and more from conservative settings of response criteria. PMID- 11405320 TI - A longitudinal study of gender differences in depressive symptoms from age 50 to 80. AB - The Obvious Depression Scale was administered to 739 community residents at ages 50, 60, and 80 years, with 151 present at all waves. Although selective attrition influenced the level of depressive symptoms in cross-sectional vs. longitudinal samples, both sets of analyses revealed higher scores in women than in men at ages 50 and 60, but not at age 80. Men showed increases in depressive symptoms from age 60 to 80, but women did not (interaction p < .002). This interaction was not present in somatic symptoms, which increased across time in both genders. Potential explanations include differential changes in social roles with aging. PMID- 11405321 TI - Perceptions of an old female eyewitness: is the older eyewitness believable? AB - Young adults rated an old female witness (82 years) to be less competent but more honest than a young female witness (28 years). The effect of age stereotyping on believability was assessed indirectly by using a variant of Loftus' misinformation paradigm. Regression analysis showed that the more competent a witness was perceived as being, the more influential was the misinformation she provided. This was true, however, only when the witness was young because the relatively lower competence of the old witness was associated with nonsignificant misinformation effects. These data suggest that negative beliefs associating incompetence with old age may compromise the believability of older eyewitnesses. PMID- 11405322 TI - Age differences in social preferences among Taiwanese and Mainland Chinese: the role of perceived time. AB - Socioemotional selectivity theory holds that as people recognize the inevitable constraint of time imposed by mortality, their social goals change, motivating them to limit social contacts to those with whom they are emotionally close. This theory was tested among Taiwanese and Mainland Chinese. As predicted, results showed that older adults (aged 60-90 years) in both cultures were more likely than younger adults (aged 18-30 years) to prefer familiar social partners who were most likely to provide emotionally close social interactions. Mainland Chinese, who as a group have shorter actuarial life expectancy, were more likely to prefer familiar social partners than were Taiwanese. These age and cultural differences were eliminated when differences in perceived time were statistically controlled for. PMID- 11405323 TI - Social perceptions of failures in memory monitoring. AB - Can repetitious verbal behavior elicit negative stereotypes about age, memory, and competence? Young adults (n = 102, M age = 19.3 years) and community-dwelling older adults (n = 98, M age = 67.3 years) read a script containing a hypothetical conversation between 2 female targets. In the experimental condition, 1 of the targets repeated several statements during the conversation. Significant Target x Condition interactions were obtained on measures of perceived age, competence, and memory ability. The results demonstrate that conversational behavior may contribute to negative age-related perceptions in the same way as other external markers of age (e.g., vocal and facial features). PMID- 11405324 TI - Parental cancer as a risk factor for brain tumors (Sweden). AB - OBJECTIVE: We used the nationwide Swedish Family-Cancer Database to analyze the risk for adult (15-61 years) brain tumors in offspring through parental cancer probands. Additionally, cancer risks were assessed among siblings of brain tumor probands. METHODS: In offspring and parents, respectively, 5,425 and 20,938 cases of brain tumors were diagnosed between the years 1958 and 1996. Groups of offspring were compared by calculating standardized incidence ratios (SIRs) for brain tumors. RESULTS: Of brain tumor patients, 2.1% had a parent with nervous system cancer; SIRs were 1.7, 2.4, and 2.5 for all brain tumors, astrocytomas, and meningiomas, respectively. Parental endometrial cancer and melanoma were associated with offspring astrocytoma, and parental breast and thyroid cancers with offspring ependymoma and neurinoma, respectively. SIR for sibling nervous system tumors from brain tumor probands was not increased overall but was 2.5 in those diagnosed at ages 15-34 years. CONCLUSION: These data show a familial risk for brain tumors among adults. PMID- 11405325 TI - Environmental risk factors for non-Hodgkin's lymphoma: a population-based case control study in Languedoc-Roussillon, France. AB - OBJECTIVE: To investigate the occupational and environmental risk factors related to non-Hodgkin's lymphoma (NHL). METHODS: A case-control study was performed during the 1992-1996 period in Languedoc-Roussillon, southern France. Four hundred and forty-five cases of histologically diagnosed NHL were declared. One thousand and twenty-five randomly selected population controls were interviewed about their medical histories; occupational exposures, such as chemicals, pesticides, and electromagnetic radiation; and toxic habits. RESULTS: The following factors were independently and significantly related to NHL as a result of the multivariate analysis: a previous hematopoietic malignancy (ORa = 11.5, 95% CI 2.4-55.4), a history of hives (ORa = 1.7, 95% CI 1.2-2.2), benzene exposure > 810 days (ORa = 4.6, 95% CI 1.1-19.2), daily welding (ORa = 2.5, 95% CI 1.2-5.0), and activity of radio operator (ORa = 3.1, 95% CI 1.4-6.6). To be an agricultural professional seemed slightly related to NHL in reference to non professionals (ORa = 1.5, 95% CI 1.0-2.1). All of these results have also been adjusted for age, gender, education level, and urban setting. CONCLUSIONS: As some of the reported associations were based on a very small proportion of exposed subjects, further investigations are necessary to confirm our results. However, the findings suggest that factors related to altered immune functions such as a history of hematopoietic malignancy, history of hives, occupational exposure to benzene, or being an agricultural professional might increase the risk of NHL. Currently, underlying mechanisms for these associations are still unclear, and further investigations focused on interactions between immunity alterations and different chemicals would be of great interest. PMID- 11405326 TI - Bone mineral density and endogenous hormones and risk of breast cancer in postmenopausal women (United States). AB - OBJECTIVE: This case-cohort study was designed to examine whether total hip bone mineral density (BMD) is independently associated with breast cancer over and above its association with other determinants, including levels of total and bioavailable estradiol and testosterone and sex-hormone binding globulin. METHODS: Our study population was selected from a cohort of 8,203 postmenopausal women who were screened for the Fracture Intervention Trial in 1992, at which time BMD was assessed, and blood samples were obtained. A total of 109 women developed breast cancer during four years of follow-up; 173 other randomly selected women from the larger cohort were also selected. Cox proportional hazards with robust variance adjustment was used for these analyses. RESULTS: Relative to women in the lower fourth of the BMD distribution, the risk associated with being in the upper fourth was 2.6 (95% confidence interval (CI) 1.1-5.8). After adjusting for serum hormone levels, the corresponding relative risk was 2.5 (95% CI 0.9-5.2). With body mass index and number of years since menopause added to the multivariate analysis, the relative risk decreased to 1.4 (95% CI 0.5-4.0). CONCLUSIONS: BMD may not influence breast cancer risk independent of its relationship with endogenous hormones and measured covariates. PMID- 11405327 TI - Relatives of prostate cancer patients have an increased risk of prostate and stomach cancers: a population-based, cancer registry study in Finland. AB - OBJECTIVE: Five to ten percent of prostate cancers may be caused by inherited genetic defects. In order to explore the nature of inherited cancer risks in the genetically homogeneous Finnish population, we investigated the incidence of prostate cancer and other cancers in first-degree relatives of prostate cancer patients by linking the population-based parish records on relatives with the Finnish Cancer Registry (FCR) data. METHODS: The study population was composed of first-degree relatives of two groups of prostate cancer patients diagnosed in Finland during 1988-1993: (1) all early-onset (<60 years) patients (n = 557) from the entire country, (2) a sample (n = 989) of prostate cancer patients diagnosed at an age of > 60 years. A total of 11,427 first-degree relatives were identified through parish records, and their cancer incidence was determined based on a total of 299,970 person-years. Standardized incidence ratios (SIR) were calculated based on expected cancer rates in the general population. RESULTS: The SIR of prostate cancer was increased in both Cohort 1 (2.5, 95% CI 1.9-3.2) and Cohort 2 (1.7, 95% CI 1.4 2.1). The risk of prostate cancer was high for relatives of patients diagnosed at an early age, and then leveled off for patients in the median age of prostate cancer diagnosis (70-79 years). However, the prostate cancer risk for relatives of patients diagnosed > or = 80 years was again statistically significantly elevated (SIR 1.8, 95% CI 1.3-2.6), suggesting a contribution of genetic factors to prostate cancer also at a late age of onset. Gastric cancer was the only other cancer type with a significantly elevated risk among the relatives. Increased risk of gastric cancer was seen only in male relatives of prostate cancer patients diagnosed at an early age, with the highest risk detected for the male relatives of prostate cancer patients diagnosed at an age of 55 years or less (SIR 5.0, 95% CI 2.8-8.2). CONCLUSIONS: Our population based study indicates that hereditary factors may play an important role in the development of prostate cancer among the relatives of men diagnosed both at younger and older ages. This finding is relevant in the context of our observations that HPCX (hereditary prostate cancer susceptibility locus on Xq27 28) linkage in Finland is found exclusively among families with late age of onset. The association of gastric cancer with prostate cancer has not been reported previously, and may reflect the effects of a novel predisposition locus, which increases the risk to both of these common tumor types. PMID- 11405328 TI - Are coffee, tea, and total fluid consumption associated with bladder cancer risk? Results from the Netherlands Cohort Study. AB - OBJECTIVES: Coffee, tea, and fluid consumption have been thought to influence bladder cancer incidence. In a large prospective study, these associations were investigated. METHODS: In 1986, cohort members (55-69 years) completed a questionnaire on cancer risk factors. Follow-up was established by linkage to cancer registries until 1992. The multivariable case-cohort analysis was based on 569 bladder cancer cases and 3,123 subcohort members. RESULTS: The incidence rate ratios (RR) for men consuming <2 cups of coffee/day was 0.89 (95% CI 0.51-1.5) using the median consumption category (4-<5 cups/day) as reference. This RR increased to 1.3 (95% CI 0.94-1.9) for men consuming >7 cups/day, although no clear dose response association was found. The RRs decreased from 1.2 (95% CI 0.56-2.7) for women consuming <2 cups of coffee/day to 0.36 (95% CI 0.18-0.72) for women consuming >5 cups/day compared to the median consumption category (3-<4 cups/day). Men and women who abstained from drinking tea had a RR of 1.3 (95% Cl 0.97-1.8) compared to those consuming 2-<3 cups of tea per day (median consumption category). The RR for men and women comparing highest to lowest quintile of total fluid consumption was 0.87 (95% CI 0.63 1.2). CONCLUSION: The data suggest a possible positive association between coffee consumption and bladder cancer risk in men and a probable inverse association in women. Tea consumption was inversely associated with bladder cancer. Total fluid consumption did not appear to be associated with bladder cancer. PMID- 11405329 TI - Cigarette smoking, alcohol consumption, and endometrial cancer risk: a population based study in Sweden. AB - OBJECTIVE: To assess effects of cigarette smoking and alcohol consumption on the risk of endometrial cancer among postmenopausal women. METHODS: We performed a nationwide population-based case-control study among postmenopausal women aged 50 74 years in Sweden, including 709 incident endometrial cancer cases and 3368 controls. RESULTS: Compared to never smokers, recent/current smokers had a decreased risk of endometrial cancer (multivariate OR 0.61, 95% CI 0.47-0.80), but former smokers presented no substantial difference in risk (multivariate OR 0.90, 95% CI 0.72-1.14). We observed a decreased risk of endometrial cancer for postmenopausal smoking, but there was no clear impact on risk for premenopausal smoking. The inverse association of smoking with risk was not explained by differences in body mass index between smokers and nonsmokers. Alcohol consumption was not clearly associated with risk of endometrial cancer. The multivariate OR for women consuming up to 1.6 g of alcohol per day was 1.12 (95% CI 0.88-1.44), and 0.92 (95% CI 0.70-1.20) for women consuming more than 4 g per day (p for trend over categories = 0.44). CONCLUSIONS: Current cigarette smoking reduces the risk of postmenopausal endometrial cancer, but the inverse association dissipates after smoking cessation. Premenopausal smoking might not affect risk of postmenopausal endometrial cancer. Alcohol consumption is not materially associated with risk. PMID- 11405330 TI - Trends in breast cancer mortality, incidence, and survival, and mammographic screening in Tuscany, Italy. AB - OBJECTIVE: The study describes breast cancer mortality trends in Tuscany (period 1970-97), comparing Florence with the rest of Tuscany (Florence excluded), and, for Florence, incidence (period 1985-94) and survival (1985-86 versus 1991-92) trends, taking into account the diffusion of screening. METHODS: Mortality and incidence rates, age-adjusted on the European population, and 95% confidence intervals (95% CI). Five-year relative survival rates and estimates of risk of dying provided by the Cox model. RESULTS: Mammographic screening, started at the beginning of the 1970s in some municipalities, largely involved the Florence area after 1990 (mammograms/years: from 8,000-9,000 to 28,000-29,000, respectively, before and after 1990). In the same period no population-based screenings were ongoing in the rest of Tuscany. A significant mortality drop was observed in Tuscany (-3.7%/year), starting at the beginning of the 1990s and observed for ages < or = 74 (especially ages 40-49: -11.2%/year). The drop was similar in Florence and in the rest of Tuscany. In ages 50-69, incidence, increasing between 1985-87 and 1988-90 (+6.5%), rose sharply in 1991-94 (+17.0%); it was stable in other ages. Local disease increased more markedly in ages 50-69 (globally: +88.3%), but also in other ages (+20-30%). Regional and metastatic cancers decreased. A significantly better 5-year survival was observed among cases diagnosed in 1991-92, persisting after adjustment by extent of disease. CONCLUSION: Even if the causes of breast cancer mortality trends are not easy to clarify in an observational study. our data suggest that the drop in mortality observed in Tuscany at the beginning of the 1990s could be largely explained by both earlier detection, outside of an organized screening program, and by better treatments. The increase in incidence and the shift in stage distribution that occurred before the enlargement of the screening area and in age groups not involved in the program, supports the role of a 'spontaneous' widespread earlier detection. The better survival of the period 1991-92, only partly explained by the shift in stage at diagnosis, indirectly supports the role of improvement in therapy. PMID- 11405331 TI - Finding incident breast cancer cases through US claims data and a state cancer registry. AB - OBJECTIVE: With the increasing availability of automated health-care data, new methods are available to screen large populations for the presence of cancer diagnoses. However, it is crucial to evaluate how completely incident cancer cases can be ascertained using these data sources. METHODS: We used capture recapture techniques to estimate the total number of incident breast cancer cases occurring within one state during a 3-year period. We then compared the ascertainment of these cases by the following two data sources: claims for breast cancer surgery recorded in Medicaid and Medicare data vs a cancer registry in the same state. RESULTS: Medicaid-Medicare breast cancer surgery claims identified 68% of the total estimated number of incident breast cancer cases while cancer registry data identified 78%. Case ascertainment improved markedly to 91% when both registry and Medicare-Medicaid data sources were used together. The sensitivity of ascertainment was lower for Medicaid-Medicare data among those aged under 65 and non-white; ascertainment was lower for the registry among women who were aged under 65, poor, and non-white. CONCLUSIONS: Combining health insurance claims data with a population-based cancer registry improved the identification of incident cases of breast cancer, and may be particularly useful among demographic groups found to be at highest risk of under-ascertainment such as younger women, the poor, and racial minorities. PMID- 11405332 TI - Risk factors for penile cancer: results of a population-based case-control study in Los Angeles County (United States). AB - The etiology of penile cancer is poorly understood, with neonatal circumcision being one of the few recognized nondemographic risk factors. Multiple logistic regression was used to analyze interview data from 100 matched case-control pairs; cases of carcinoma in situ (CIS) and invasive carcinoma of the penis were analyzed separately as well as together. Phimosis was strongly associated with invasive carcinoma (adjusted odds ratio [OR] = 16; 95% confidence interval [CI] = 4.5-57) but not CIS (OR = 1.7; 95% CI = 0.32-7.8), and these associations persisted when the analyses were restricted to uncircumcised subjects. Neonatal circumcision was inversely associated with invasive carcinoma (OR = 0.41; 95% CI = 0.13-1.1) but not CIS, and the observed association with invasive carcinoma was weakened appreciably when the analysis was restricted to subjects with no history of phimosis (OR = 0.79; 95% CI = 0.29-2.6). Other factors positively associated with invasive carcinoma or CIS or both were injury to the penis, cigarette smoking, physical inactivity and, to a lesser extent, genital warts and other infections or inflammation of the penis. CONCLUSIONS: Although many effects were imprecisely estimated in this study, the protective effect of circumcision on invasive penile cancer appears to be mediated in large part by phimosis; furthermore, the effects of certain factors such as phimosis and circumcision appear to differ for CIS and invasive carcinoma. PMID- 11405333 TI - Light in the built environment: potential role of circadian disruption in endocrine disruption and breast cancer. AB - Life in industrialized societies is primarily life inside buildings. Illumination from electric lighting in the built environment is quite different from solar radiation in intensity, spectral content, and timing during the 24-hour daily period. Humans evolved over millions of years with the day-night pattern of solar radiation as the primary circadian cue. This pattern maintained a 24-hour rhythm of melatonin release, as well as a host of other physiological rhythms including the sleep-wake cycle. Electric lighting in the built environment is generally more than sufficient for visual performance, but may be inappropriate for the maintenance of normal neuroendocrine rhythms in humans; e.g., insufficient during the day and too much at night. Lighting standards and engineering stress visual performance, whereas circadian function is not currently emphasized. The molecular biological research on the circadian clock and on mechanisms of phototransduction makes it clear that light for vision and light for circadian function are not identical systems. In particular, if electric lighting as currently employed contributes to 'circadian disruption' it may be an important cause of 'endocrine disruption' and thereby contribute to a high risk of breast cancer in industrialized societies. PMID- 11405334 TI - Changes in clinical practice with the unravelling of diseases: connective-tissue disorders. AB - Unravelling of diseases is achieved in steps by sequentially describing their phenotype, natural course, aetiology and pathogenesis. Through succinct clinical observation, conglomerates of heterogeneous connective-tissue disorders, such as various forms of disproportionate dwarfism, have been split into well-defined entities. They have often been confirmed by biochemical and molecular analysis. On the other hand, seemingly disparate disorders have been shown to be pathogenetically related and to be variable expressions of common defects. Examples are the mucopolysaccharidoses and type II collagenopathies. Disease recognition through splitting and lumping has improved prevention and prognostication. It is the basic requirement for future therapeutic attempts on a pathogenetic or molecular level. PMID- 11405335 TI - The use of functional genomics in C. elegans for studying human development and disease. AB - The 100 Mb Caenorhabditis elegans genome sequence is the first animal genome to be sequenced in its entirety. Many reverse-genetics tools have been developed to mine the genome sequence and to facilitate the jump between the identification of a gene sequence and the understanding of its function. Here we discuss how C. elegans can contribute to understanding of the function of genes involved in human development and disease. PMID- 11405336 TI - Applications of mass spectrometry in the study of inborn errors of metabolism. AB - During the twentieth century, and particularly in its last decade, there have been major advances in mass spectrometry (MS). As a result, MS remains one of the most powerful tools for the investigation of genetic metabolic disease. Analysis of organic acids by gas chromatography-mass spectrometry (GC-MS) and analysis of acylcarnitines by tandem mass spectrometry are still leading to the discovery of new disorders. Tandem mass spectrometry is increasingly being used for neonatal screening. New methods for lipid analysis have opened up the fields of inborn errors of cholesterol synthesis, of bile acid synthesis and ofleukotriene synthesis. The latest developments in MS allow it to be used for determination of the amino acid sequence and posttranslational modifications of proteins. There are still some major hurdles to be overcome, but soon it should be possible to detect mutant proteins directly rather than by cDNA or genomic DNA analysis. Measurement of which proteins are overexpressed and underexpressed ('proteomics') should provide further information on the pathogenesis of complications of inborn errors, e.g. hepatic cirrhosis. The use of stable isotopes in conjunction with MS allows us to probe metabolic pathways. As an example, evidence is presented to support the contention that vitamin E and its oxidation product are catabolized by peroxisomal beta-oxidation. Mass spectrometry also has a major role in monitoring new forms of treatment for inborn errors. PMID- 11405337 TI - Clinical, biochemical and genetic aspects and neuronal migration in peroxisome biogenesis disorders. AB - Peroxisome biogenesis disorders (PBDs) are severe autosomal recessive neurological diseases caused by a defect of peroxisomal assembly factors. Zellweger syndrome, the most severe phenotype, is characterized by hypotonia, psychomotor retardation and neuronal migration disorder. Neonatal adrenoleukodystrophy and infantile Refsum disease are milder phenotypes of this disease. Thirteen complementation groups have been established since the genetic heterogeneity of PBDs was elucidated in 1988. Eleven genes for PBDs have been identified either by a functional complementation cloning or by EST homology searches. In 1992, the first gene for PBDs, PEX2, was identified. It encodes peroxisomal integral membrane protein with a RING finger domain. PEX5 and PEX7 are the genes for peroxisomal targeting signal (PTS)-1 and -2 receptors, respectively. PEX3, PEX16 and PEX19 are considered to be required for the early stage of peroxisome biogenesis. PEX13 protein has an SH3 docking site that binds to the PTS-1 receptor. PEX1 and PEX6 encode ABC protein, and PEX10 and PEX12 also encode integral membrane protein, with RING finger. Temperature-sensitivity, whereby peroxisomal biogenesis and metabolic dysfunctions are restored at 30 degrees C in cells from mild phenotypes, is a useful event for predicting the clinical severity and for elucidation of peroxisome biogenesis. Investigations using knockout mice are expected to facilitate understanding of migration disorders. PMID- 11405339 TI - Clinical and molecular studies of mitochondrial disease. AB - Our knowledge of mitochondrial disorders has expanded enormously through advances in mitochondrial DNA (mtDNA) research during the last decade. This has also provided information about the complicated genotype-phenotype relationship based on distinct characteristics of mtDNA itself. These include multicopy, vulnerability to the mutation, and cytoplasmic inheritance. There are complications of heteroplasmy, tissue/cell specificity, accumulation of mutations and material inheritance. PMID- 11405338 TI - Import of proteins into mitochondria: a novel pathomechanism for progressive neurodegeneration. AB - The vast majority of mitochondrial proteins are encoded as precursors by the nuclear genome. A major aspect of mitochondrial biogenesis is therefore the transfer of nuclear-encoded, cytosplasmically synthesized precursor proteins across and into the mitochondrial membranes. During the past years the use of simple model organisms such as the yeasts S. cerevisiae and N. crassa has helped considerably to identify and unravel the structure and function of a substantial number of components involved in targeting of nuclear-encoded preproteins to mitochondria. Several pathways and a number of components were characterized that are involved in guiding mitochondrial preproteins to their specific sites of function. In particular, import of nuclear-encoded precursor proteins into and across the mitochondrial inner membrane is mediated by two distinct translocases, the TIM23 complex and the TIM22 complex. Both TIM complexes cooperate with the general preprotein translocase of the outer membrane, TOM complex. The TIM complexes differ in the their substrate specificity. While the TIM23 complex mediates import of preproteins with a positively charged matrix targeting signal, the TIM22 complex facilitates the insertion of a class of hydrophobic proteins with internal targeting signals into the inner membrane. Most recently the rapid progress of research has allowed elucidation of a new mitochondrial disease on the molecular level. This rare X-linked progressive neurodegenerative disorder, named Mohr-Tranebjaerg (MT syndrome), is caused by mutations in the DDP1 gene and includes sensorineural deafness, blindness, mental retardation and a complex movement disorder. The analysis of the novel pathomechanism is based on the homology of the affected DDP1 protein to a family of conserved yeast components acting along the TIM22 pathway. This contribution briefly summarizes the current knowledge of the pathways of protein import and proposes a mechanism to explain how defective import leads to neurodegeneration. PMID- 11405340 TI - The role of chaperone-assisted folding and quality control in inborn errors of metabolism: protein folding disorders. AB - Molecular chaperones are present in the various compartments of the cell and assist the folding of newly synthesized proteins. Compared to wild-type proteins, missense mutant proteins are generally synthesized in a normal fashion, but may be impaired in their folding. A broad array of diseases that are due to misfolding of mutant proteins may be labelled conformational diseases: aggregation diseases, such as Alzheimer disease; diseases caused by negative dominance from misfolded structural proteins, such as hypertrophic cardiomyopathy; and disorders where the misfolded protein is degraded by intracellular proteases. Many metabolic disorders belong to this last category, where the so-called protein quality control systems, comprising chaperones and proteases, attempt to eliminate folding intermediates or misfolded proteins. On the basis of in vitro experiments with a limited number of missense mutations identified in patients with phenylalanine hydroxylase and fatty acid oxidation deficiencies, we discuss the cellular fate of missense mutant proteins. We find that the balance between folding to functional conformers, retention (holding) and degradation of folding intermediates or misfolded proteins is dependent on the nature of the mutation and on the efficiency of the quality control. For example, low temperature may promote formation of functional conformers, while elevated temperature usually promotes retention and degradation. We conclude that disorders caused by many missense mutations are complex diseases in which the mutation itself is a necessary major primary component, but that its effect may be modified by cellular conditions and possibly by genetic variations in the quality control systems. We suggest that this new knowledge about cell handling may open new avenues of understanding of the cell pathology and treatment of patients with metabolic disorders. PMID- 11405341 TI - A structural hypothesis for BH4 responsiveness in patients with mild forms of hyperphenylalaninaemia and phenylketonuria. AB - Deficiencies in the human enzyme phenylalanine hydroxylase (PAH) due to mutations in the PAH gene (PAH) result in the inborn error of metabolism phenylketonuria (PKU). The clinical symptom of this disease is an elevated concentration of L phenylalanine (L-Phe) in blood serum. To prevent mental retardation due to the buildup of neurotoxic metabolites of L-Phe, patients with severe PKU must be treated with a low-L-Phe diet starting early in their life. Owing to extensive newborn screening programmes and genotyping efforts, more than 400 different mutations have been identified in the PAH gene. Recently, there have been several reports of PKU patients showing a normalization of their L-Phe concentrations upon oral administration of the natural cofactor to PAH, (6R)-L-erythro-5,6,7,8 tetrahydrobiopterin (BH4). In an attempt to correlate the clinical responsiveness to BH4 administration with PKU genotype, we propose specific structural consequences for this subset of PAH mutations. Based on the location and proximity of this subset of mutations to the cofactor-binding site in the three dimensional structure of PAH, a hypothesis for BH4 responsiveness in PKU patients is presented. It is believed that some of these mutations result in expressed mutant enzymes that are Km variants (with a lower binding affinity for BH4) of the standard PAH enzyme phenotype. Oral administration of excess BH4 thus makes it possible for these mutant enzymes to suppress their low binding affinity for BH4, enabling this subset of PAH mutations to perform the L-Phe hydroxylation reaction. Most of the BH4-responsive PAH mutations map to the catalytic domain of PAH in either of two categories. Residues are located in cofactor-binding regions or in regions that interact with the secondary structural elements involved in cofactor binding. Based on the series of known mutations that have been found to be responsive to BH4, we propose that other subsets of PAH mutations will have a high likelihood of being responsive to oral BH4 administration. PMID- 11405342 TI - Liver repopulation for the treatment of metabolic diseases. AB - Orthotopic liver transplantation is the treatment of choice for several inborn errors of metabolism. Unfortunately, the supply of donor organs is limiting and therefore many patients cannot benefit from this therapy. In contrast, hepatocyte transplantation could potentially overcome the shortage in donor livers by use of cells from a single donor for multiple recipients. In classic hepatocyte transplantation, however, only 1% of the liver mass or less can be replaced by donor cells. Recently, though, it has been shown in animal models that >90% of host hepatocytes can be replaced by a small number of transplanted donor cells in a process we term 'therapeutic liver repopulation'. This phenomenon is analogous to repopulation of the haematopoietic system after bone marrow transplantation. Liver repopulation occurs when transplanted cells have a growth advantage in the setting of damage to recipient liver cells. It has been discovered that transplanted cells from extrahepatic sources such as the adult pancreas or bone marrow can also be used for liver repopulation. Because bone marrow donors are widely available, this finding raises the hope of therapeutic application of these cells in the future. Here, the current knowledge regarding therapeutic liver repopulation and the hopeful implications for treatment of liver diseases will be discussed. PMID- 11405343 TI - Enzyme replacement therapy in mucopolysaccharidosis type I: progress and emerging difficulties. AB - Mucopolysaccharidosis type I is due to a deficiency of the lysosomal enzyme alpha L-iduronidase (EC 3.2.1.76) and is associated with a defect in the catabolism of the glycosaminoglycans heparan and dermatan sulphate. The clinical picture produced by this abnormal storage is diverse and ranges from a disorder that is fatal in the early months of life, due to cardiomyopathy, to a condition compatible with a normal life-span. It has become usual to describe the phenotypes associated with this spectrum of disorders after their eponymous names, Hurler (MPS IH, severe), Scheie (MPS IS, mild) and Hurler/Scheie (MPS IH/S, intermediate). Severely affected patients have progressive learning difficulties, facial and skeletal deformities, cardiac disease, corneal clouding, respiratory compromise and joint stiffness. Patients with MPS IH typically die in the first decade of life. MPS IH/S usually have normal intelligence and die in their twenties of cardiorespiratory disease. Patients with MPS IS may have joint stiffness, aortic valve disease and corneal clouding, but are often able to live a normal life-span. Enzyme replacement therapy has been developed as a potential therapy for some patients with MPS I. This process has been helped by the study of a naturally occurring canine model of the disease, which produces a phenotype similar to MPS IH/S in the human. This review details the progress that has been made in this area and also highlights some potential problems with the introduction of therapy. PMID- 11405344 TI - Enzyme replacement and beyond. AB - During the last decade, enzyme replacement therapy for lysosomal storage diseases became a reality with the demonstration of its safety and effectiveness in type 1 Gaucher disease. Currently, enzyme replacement and several other potential therapeutic strategies are being developed for selected lysosomal storage diseases, including Fabry disease due to the deficient activity of alpha galactosidase A (alpha-Gal A). The development and clinical evaluation of these new therapies require a stepwise process, each step being rigorously reviewed and approved by national or international regulatory agencies. For lethal disorders that affect small populations, such as many inherited metabolic diseases, this process can be accelerated by 'orphan drug' and 'fast track' regulations. As an example of the drug development process, the development of recombinant human alpha-Gal A (r-halphaGal A) replacement for Fabry disease is presented, including the preclinical studies in the 'Fabry mouse' model, and the clinical phase 1/2, phase 3, and phase 3 extension studies, which demonstrate the safety and efficacy of this new therapy. PMID- 11405345 TI - Enzyme therapy for pompe disease with recombinant human alpha-glucosidase from rabbit milk. AB - Pompe disease is a metabolic myopathy caused by deficiency of lysosomal acid alpha-glucosidase. In this report we review the first 36 weeks of a clinical study on the safety and efficacy of enzyme therapy aimed at correcting the deficiency. Four patients with infantile Pompe disease were enrolled. They received recombinant human alpha-glucosidase from transgenic rabbit milk. The product is generally well tolerated and reaches the primary target tissues. Normalization of alpha-glucosidase activity in skeletal muscle was obtained and degradation of PAS-positive material was seen in tissue sections. The clinical condition of all patients improved. The effect on heart was most significant, with an impressive reduction of the left ventricular mass index (LVMI). Motor function improved. The positive preliminary results stimulate continuation and extension of efforts towards the realization of enzyme therapy for Pompe disease. PMID- 11405346 TI - Inhibition of substrate synthesis as a strategy for glycolipid lysosomal storage disease therapy. AB - The glycosphingolipid (GSL) lysosomal storage diseases are caused by mutations in the genes encoding the glycohydrolases that catabolize GSLs within lysosomes. In these diseases the substrate for the defective enzyme accumulates in the lysosome and the stored GSL leads to cellular dysfunction and disease. The diseases frequently have a progressive neurodegenerative course. The therapeutic options for treating these diseases are relatively limited, and for the majority there are no effective therapies. The problem is further compounded by difficulties in delivering therapeutic agents to the brain. Most research effort to date has focused on strategies for augmenting enzyme levels to compensate for the underlying defect. These include bone marrow transplantation (BMT), enzyme replacement and gene therapy. An alternative strategy that we have been exploring is substrate deprivation. This approach aims to balance the rate of GSL synthesis with the impaired rate of GSL breakdown. The imino sugar N-butyldeoxynojirimycin (NB-DNJ) inhibits the first step in GSL biosynthesis and has been used to evaluate this approach. Studies in an asymptomatic mouse model of Tay-Sachs disease have shown that substrate deprivation prevents GSL storage in the CNS. In a severe neurodegenerative mouse model of Sandhoff disease, substrate deprivation delayed the onset of symptoms and disease progression and significantly increased life expectancy. Combining NB-DNJ and BMT was found to be synergistic in the Sandhoff mouse model. A clinical trial in type I Gaucher disease has been undertaken and has shown beneficial effects. Efficacy was demonstrated on the basis of significant decreases in liver and spleen volumes, gradual but significant improvement in haematological parameters and disease activity markers, together with diminished GSL biosynthesis and storage as determined by independent biochemical assays. Further trials in type I Gaucher disease are in progress; studies are planned in patients with GSL storage in the CNS. PMID- 11405347 TI - Rare diseases and the assessment of intervention: what sorts of clinical trials can we use? AB - There is increasing emphasis on the importance of practising evidence-based medicine. Randomized controlled trials are the standard way to assess the benefits of an intervention, and observational studies are not usually accorded much weight; the results are likely to be considered misleading. For rare diseases, there are great difficulties in obtaining adequate evidence for interventions or for the benefits of early diagnosis. This is because the disorders are not only very rare but also have variable expression, may have very long courses, and have incompletely known late effects; and surrogate end-points often have to be used. Randomized controlled trials are usually impossible because of inadequate power, and because there are preconceived notions of the effects of treatments already in use. The adoption of the best possible design for observational trials, formation of a central registry of such trials, and a greater general appreciation of the problems that rare diseases pose will help in obtaining the best possible evidence for the effects of interventions. PMID- 11405348 TI - Newborn mass screening versus selective investigation: benefits and costs. AB - Cost-benefit analysis of newborn screening has an unimpressive record and yet it is still regarded as an important decision tool. This workshop surveyed ongoing research into the costs and benefits of systematic whole-population screening, as opposed to selective investigation of symptomatic patients, for inherited metabolic disease. Much current interest is focused on newborn screening by tandem mass spectrometry, which can replace current methods for detecting phenylketonuria and cover a much wider range of diseases. Two observational studies are comparing cost-effectiveness of tandem mass spectrometry screening versus symptomatic diagnosis in either concurrent or historical control populations. A number of other studies are assessing screening performance against predetermined criteria but without any formal control group. Medium-chain acyl-CoA dehydrogenase deficiency is the most common of the additional diseases being detected and it seems that octanoylcarnitine in blood is a particularly sensitive indicator: some of the cases detected by screening have genotypes suggesting a relatively low risk of serious metabolic decompensation. Ongoing studies should provide further quantitative and qualitative data but will not in themselves define the optimum balance between screening sensitivity and specificity. PMID- 11405349 TI - New England Consortium: a model for medical evaluation of expanded newborn screening with tandem mass spectrometry. PMID- 11405350 TI - Amino acids: analytical aspects. AB - This workshop was organized as a direct response to concerns and queries raised by laboratory personnel, both in Europe and in the United States, about the imminent withdrawal of Beckman Coulter from the amino acid analysis market. The topics covered included external quality control schemes, standard operating procedures for amino acid analysis and instrumentation, both from a user's perspective and that of the company representatives. There was a discussion panel of all speakers following the presentations. PMID- 11405351 TI - The management of organic acidaemias: the role of transplantation. AB - This workshop addressed the issue of treatment in propionic acidaemia (PA) and methylmalonic acidaemia (MMA) and in particular the outcome of conventional management compared with organ transplantation. Although it appears that with medical treatment the mortality for early-onset disease has improved, long-term outcome remains poor. However, liver and liver/kidney transplantation is associated with a high perioperative mortality and long-term complications. It offers only a partial cure. The indications for transplantation remain unclear. It is recommended that a register of PA and MMA patients should be established. PMID- 11405352 TI - Diagnosis of mitochondrial disorders: clinical and biochemical approach. AB - The topic of Workshop W3-1 was clinical and biochemical approaches to the diagnosis of mitochondrial respiratory chain disorders. Four main questions were addressed in an attempt to make some progress towards a consensus diagnostic approach: What are the major limitations in diagnosis of respiratory chain dysfunction? What is the ideal approach to investigating children with a suspected respiratory chain disorder? Can we begin to develop consensus diagnostic criteria? Can we develop a quality assurance (QA) scheme for respiratory chain enzyme assays? The workshop demonstrated strong consensus on recognizing the limitations of current diagnostic approaches, on the ideal diagnostic approach and on the desirability of an enzyme QA scheme. There was also support for the desirability of consensus diagnostic criteria, albeit with some concerns about the practicality of gaining consensus. Two potential approaches to developing consensus criteria were described. PMID- 11405353 TI - Garrod's foresight; our hindsight. AB - Archibald Edward Garrod introduced a paradigm, new for its day, in medicine: Biochemistry is dynamic and different from the static nature of organic chemistry. It led him to think about metabolic pathways and to recognize that variation in Mendelian heredity could explain an 'inborn error of metabolism'. At the time, Garrod had no idea about the nature of a gene. Genes are now well understood, genomes are being described for one organism after another (including H. sapiens) and it is understood that genomes 'speak biochemistry (not phenotype)'. Accordingly, in the era of genomics, biochemistry and physiology become the bases of functional genomics and it is possible to appreciate why 'nothing in biology makes sense without evolution' (and nothing in medicine will make sense without biology). Mendelian, biochemical and molecular genetics together have revealed what lies behind the four canonical inborn errors described by Garrod (albinism, alkaptonuria, cystinuria and pentosuria). Both older and newer ideas in genetics, new tools for applying them, and renewed respect for the clinician-scientist will enhance our understanding of the human biological variation that accounts for variant states of health and overt disease; an 'unsimple' phenotype (phenylketonuria) is used to illustrate in some detail. What can be known and what ought to be done with knowledge about human genetics to benefit individuals, families and communities (society) is both opportunity and challenge. PMID- 11405354 TI - Cryogen spray cooling and pulsed dye laser treatment of cutaneous hemangiomas. AB - When a cryogen spurt is applied to the skin surface for tens of milliseconds, cooling remains localized in the epidermis, leaving the temperature of deeper hemangioma vessels unchanged. The objective of this study was to compare the efficacy and safety of noncooled laser treatment (NC-LT) vs. cryogen spray cooling plus laser treatment (CSC-LT) for cutaneous hemangiomas in a large series of patients. A retrospective review was conducted of 164 patients treated with the pulsed dye laser (lambda = 585 nm; taurho = 450 microsec) over an 8-year period. Eighty-two patients received NC-LT using light doses of 5.5 to 8 J per square centimeter. Subsequently, 82 patients received CSC-LT using light doses of 9 to 10 J per square centimeter. The primary efficacy measure was quantitative assessment of improvements in lesional volume, texture, and color. Safety was evaluated for each treatment group by monitoring for adverse effects. Based on chi-squared analysis, there were clinical and significant differences in the number of treatments (p = 0.001), and improvement in volume (p = 0.008) and texture (p = 0.001) of the CSC-LT group compared with the NC-LT group. Permanent adverse effects were not observed in either group. In conclusion, CSC permitted the use of higher incident light doses for treatment of cutaneous hemangiomas, resulting in fewer treatments required and better improvement in lesional volume and texture. PMID- 11405355 TI - The maximally vascularized central pedicle breast reduction: evolution of a technique. AB - A retrospective review of patient medical records was used to evaluate a surgical procedure for reduction mammaplasty. Information obtained during this review was then used to determine areas of possible improvement in patient selection and in surgical technique. Modifications were made, and a second series of patients was treated according to the new selection criteria, and by a modified surgical technique. The medical records of the second series of patients were reviewed. The two series of patients are compared. The initial series includes a retrospective review of data collected on 43 consecutive patients treated for bilateral macromastia using the central pedicle breast reduction technique of Hester and colleagues. The results of this initial series of patients were reviewed and presented at the Northwest Society of Plastic Surgeons Meeting; Lanai, HI, in 1995. Data obtained from that review, resulted in modifications in patient selection criteria and in operative technique by the senior author (RPR). An additional 110 consecutive patients have been treated using this modified protocol. The authors review the same data for the second series of patients and compare the results of the two series. They illustrate the use of the central pedicle technique in 153 patients. They have used the central pedicle reduction mammaplasty technique of Hester and colleagues as a model to demonstrate evolution in a surgical technique after evaluation of data from the medical record. A 50% reduction in complications and a 35% reduction in operative time were obtained. PMID- 11405356 TI - Intraoperative evaluation of blood flow in the internal mammary or thoracodorsal artery as a recipient vessel for a free TRAM flap. AB - Although the free microvascular transverse rectus abdominis musculocutaneous (TRAM) flap is in routine use for breast reconstruction, little is known of its hemodynamics. The purpose of this study was to determine whether any differences exist when the free TRAM flap is anastomosed to the thoracodorsal or internal mammary vessels. The study comprised 25 patients receiving a free TRAM flap for breast reconstruction. The thoracodorsal vessels were used as recipients in 21 patients and the internal mammary vessels were used in 4 patients. Blood flow rate was measured directly in the donor and recipient arteries, and after anastomosis by a transit-time ultrasonic flowmeter (CardioMed). Two- and 3-mm probes were used. The blood flow rate in the donor artery (deep inferior epigastric) before flap dissection was 11 +/- 6 ml per minute (mean +/- standard deviation). The rate was significantly (p < 0.05) lower (5 +/- 3 ml per minute) in the recipient thoracodorsal artery than in the donor, but after transplantation it increased to 14 +/- 5 ml per minute (p < 0.05), attaining the same value as the donor artery. The blood flow rate in the intact internal mammary artery was significantly higher (25 +/- 10 ml per minute) than in the donor and thoracodorsal arteries, but after anastomosis it dropped to the same value (12 +/- 3 ml per minute; p < 0.05) as the donor artery. The intake of blood in TRAM flaps supplied by the intemal mammary artery seems to be no greater than that in free flaps anastomosed to thoracodorsal vessels, although the flow in the internal mammary artery was much higher. The authors concluded that the blood supply in a free TRAM flap is independent of the flow in the recipient artery and that thoracodorsal vessels, although often in a scarred bed and radiated, are as suitable for anastomosing a free TRAM flap as are internal mammary vessels. PMID- 11405357 TI - Continuous quality management of breast hypertrophy by the German Association of Plastic Surgeons: a preliminary report. AB - In 1998, the German Association of Plastic Surgeons started a new quality assurance program based on a standardized questionnaire to examine the tracer diagnosis "breast hypertrophy." The authors present the results of a pilot study evaluating 799 patients (mean age, 38 years). Breast reduction was performed as an inpatient procedure in 99% of patients. Mean weight reduction was 1,135 g (range, 140-3,870 g). The inverted-T scar technique was used in 45% of patients, the vertical scar technique in 52% of patients, and other techniques were restricted to 2% of patients. The selection of technique was based on the choice of the surgeon. The overall incidence of complications was 21.5%, and included predominantly wound dehiscence, without marked differences between the two techniques. Less than 10 years after the introduction of a new technique for breast reduction with reduced length of scar, it is now used as often as traditional methods. PMID- 11405358 TI - Thrombolytic therapy: what is its role in free flap salvage? AB - Thrombolytic agents have been demonstrated to improve free flap salvage in animal models. However, clinical evidence regarding their efficacy has been scant. The authors reviewed their experience with flap salvage using thrombolytic therapy in 1,733 free flaps from February 1990 to July 1998. Patients with intraoperative pedicle thrombosis were excluded from this review. Forty-one of the 55 free flaps that were reexplored emergently were identified as having pedicle thrombosis. Of these 41 flaps, 28 free flaps were salvaged (flap salvage group, 68%) and 13 free flaps failed (flap failure group, 32%). Thrombolytic therapy (urokinase in 7 patients, tissue plasminogen activator in 1 patient) was used in six flaps in the flap salvage group and two flaps in the flap failure group. Statistical analysis demonstrated no difference between the two groups with regard to thrombolytic therapy. There was also no difference between the two groups with regard to use of systemic heparin (100-500 U per hour) at the time of pedicle thrombosis or with regard to whether Fogarty catheters were used. Smoking, preoperative radiotherapy, and the use of interpositional vein grafts during initial flap reconstruction had no impact on the outcome of flap salvage. The flap salvage group was reexplored at a mean of 1.5 days compared with the flap failure group, which was reexplored at a mean of 4.2 days (p = 0.007). Early detection of pedicle thrombosis remains the most important factor in the salvage of free flaps. Although these numbers are small and definitive statements cannot be made, the role of thrombolytic agents in free flap salvage requires further clinical evaluation. PMID- 11405359 TI - Fingertip replantation at or distal to the nail base: use of the technique of artery-only anastomosis. AB - The authors describe the functional and aesthetic results of microsurgical replantation of 21 fingertip amputations at or distal to the nail base-namely, zone I amputations. There were 15 male and 6 female patients, with an average age of 26 years (age range, 1-41 years). Replantations were performed using the anastomosis of the artery-only technique, with neither vein nor nerve repair. Venous drainage was provided by an external bleeding method with a fish-mouth incision in "distal" zone I amputations for approximately 7 days, and by the use of leeches in more "proximal" zone I amputations for 10 to 12 days. Results indicated that the overall survival rate was 76%, with 16 of 21 digits surviving. Sensory evaluation at an average follow-up of 12 months (range, 6-18 months) revealed an average static two-point discrimination of 6.1 mm (range, 2.0-8.0 mm). Considering the unfavorable results and the donor site morbidity of various fingertip reconstructions, a microsurgical fingertip replantation should always be considered except in extremely distal, clean-cut, pediatric cases, in which case a composite graft is a possibility. The results of this series indicate that an amputated fingertip in zone I can be salvaged successfully by microvascular anastomosis of the artery only, with a nonmicrosurgical method of venous drainage. Furthermore, acceptable sensory recovery can be expected without any nerve coaptation. PMID- 11405360 TI - Walk-through injuries: glass door facial injuries. AB - During 1998, 13 patients were treated in the Tel-Aviv Sourasky Medical Center for complex facial soft-tissue injuries caused by passing through large, clear glass doors. All epidemiological details were gathered and analyzed. Of 1,100 soft tissue facial injury admissions in 1 year, 13 patients had a substantial soft tissue facial injury after passing through a glass barrier. Nine were injured during leisure time activity, five in a shopping mall, and four in their residence. Interestingly, the authors found a common pattern of facial injuries in all patients. It consisted of large, irregular, composite skin and soft-tissue flaps as well as large, tom, irregular skin lacerations. The nose was injured predominantly, and the injury was particularly complex. Their recommended management of these injuries is a thorough and careful evaluation of flap viability. Surgical management of avulsed, viable flaps includes margin debridement and repositioning. If the flap is narrow enough, it can be debrided and the margins adapted primarily. If viability of part of the flap is in doubt, that part should be debrided and used as a composite graft. When this graft dies, a full-thickness graft is taken from another facial site. The cosmesis of such a graft is better than using the debrided, thin segment as a skin graft that is too thin. The authors emphasize that there is a need to encourage authorities to reinforce regulations relating to injury prevention from architectural glass. The first is to use special glazing, either tempered glass, laminated glass, or both. The other method of improving safety is by indicating glass using decorations or warning stickers, or by making it partly translucent. Unless these regulations are obeyed, fatal or complex trauma may occur. PMID- 11405361 TI - Facilitated tissue expansion with topical estriol. AB - Tissue expansion is a helpful technique in reconstructive plastic surgery. Unfortunately, tissue expansion still needs to be improved. Twenty-four male Wistar rats were used to evaluate the effect of estriol on tissue expansion. The agents hyaluronidase, estriol, and base cream (as a control) were applied topically to separate animal groups for 5 weeks, and their effects were studied on tissue expansion. Both hyaluronidase (p < 0.05) and estriol (p < 0.001) enhanced the rate of expansion when compared with control animals. Estriol was more effective than hyaluronidase (p < 0.05). Breaking strengths were measured in the estriol and the control groups. Breaking strength was not evaluated in the hyaluronidase group because of the necrotic changes seen at the end of the fifth week. The breaking strength was higher in the control group than in the estriol group (p < 0.05). The authors suggest that topical estriol be used as an adjunctive agent to facilitate tissue expansion. PMID- 11405362 TI - Using lower eyelid fascial slings for recalcitrant burn ectropion. AB - Burns of the lower eyelid represent a difficult management problem. Even with skin grafting, scarring and contraction can result in ectropion. This condition creates a marked aesthetic deformity, poses a risk of corneal exposure, and jeopardizes the patient's vision. Historically, full-thickness skin grafts and tarsorrhaphies have been described to manage recurrent ectropion. The lower eyelid fascial sling (LEFS) uses a temporalis fascial strip to create a suspension sling for the lower eyelid. In a series of 7 patients, the LEFS procedure resolved the symptoms permanently in all patients. Because the LEFS procedure counteracts the natural scar contraction forces of lower eyelid burns, it can resolve recalcitrant ectropion successfully while preserving the function and aesthetics of the lower eyelid. PMID- 11405363 TI - Lack of correlation between objective and subjective evaluation of residual stigmata in cleft patients. AB - The multidisciplinary cleft team of the Academisch Ziekenhuis Vrije Universiteit treats cleft patients until they are 18 years old, when treatment is supposedly complete, with patients having a minimum of residual facial stigmata. The authors evaluated the relationship between objective residual facial deformities present and the patients' subjective judgment of the result of treatment by interviewing 50 patients (age range, 19-25 years) regarding their facial appearance and possible remaining wishes for secondary correction. All patients were subjected to 13 in vivo anthropometric measurements, and 5 inclinations were assessed on lateral facial photographs. The results of these measurements were compared with normal values to establish possible objective deformities. The subjective and the objective results were compared for each individual. Anthropometric evaluation revealed an average of 5.3 divergent measurement values per patient (range, 1-8 divergent values). No correlation was found between objective judgment and the patient's subjective judgment. A weak negative correlation could be demonstrated between the score on a visual analog scale and the wish for secondary correction. Surgeons should be aware that cleft patients have no anthropometric interest in their facial appearance. PMID- 11405364 TI - The preputium: an overlooked skin graft donor site. AB - Plastic surgeons often encounter tissue defects consisting of the superficial layer of the skin. Fortunately, many of these defects are reconstructed easily with skin grafts. The authors have used the preputium as a skin graft for the reconstruction of skin defects of various sizes and locations in children. The defect size ranged from 2.3 x 4.0 cm to 4.8 x 6.5 cm. The average defect size was 3.5 x 5.5 cm. The defects were caused from trauma, acute burn, and release of burn contractures, and were located in the lower extremity, upper extremity, and scalp. All patients were treated successfully. No complications of the donor area or recipient area were seen. The prepuce should be included as an additional tool in the reconstructive surgeons armamentarium because of its many advantages, including easy harvest, high viability and elasticity, and low donor area morbidity. PMID- 11405365 TI - Prenatal exposure to nitrofen induces Fryns phenotype in mice. AB - Prenatal exposure to nitrofen is known to cause multiple malformations in mice. The reported malformations include lung hypoplasia, diaphragmatic hernia, cardiovascular defects, skeletal malformations, cleft palate, and renal abnormalities. The authors present detailed findings of craniofacial defects after prenatal exposure to nitrofen, and propose that together with the previously reported malformations, nitrofen exposure induces a Fryns phenotype in mice. Fryns syndrome is a rare human genetic syndrome that is an autosomal recessive disorder characterized by lung hypoplasia, diaphragmatic hernia, craniofacial malformations, skeletal malformations, cardiovascular malformations, and genitourinary malformations. Timed-pregnant Swiss Webster mice were gavage fed 25 mg of nitrofen on day 8 of gestation. Control animals received olive oil. Osteogenesis and chondrogenesis were studied in fetuses recovered on day 17 after Alcian blue-Alizarin red staining. Approximately 26% of the nitrofen-exposed embryos had severe craniofacial defects, and there was generalized delay in chondrogenesis and osteogenesis throughout the skeleton. No such defects were noted in the control group. The authors propose that prenatal exposure to nitrofen induces a Fryns phenotype in mice, and thus speculate that nitrofen may target similar molecular mechanisms to those that lead to Fryns syndrome. PMID- 11405366 TI - Successful breast reconstruction with a perforator to deep inferior epigastric perforator flap. AB - The authors report their experience with deep inferior epigastric perforator (DIEP) flap breast reconstruction in which an unusual recipient site was used. Successful anastomosis between a suitable perforating vessel from the internal mammary axis and the deep inferior epigastric bundle was performed, and the advantages of this alternative recipient site (perforator to the DIEP flap) are examined. PMID- 11405367 TI - Two-year histological outcome of facial lipofilling. AB - Although there are numerous clinical reports relating to the injection of adipose tissue fragments, only few histological assessments of the histology of these grafts in late follow-up have been made. The authors report a case of adipose tissue harvested with a sharp cannula using low-pressure suction and injection in the lower eyelid areas with subsequent sagging of the graft, although maintenance of volume, after 2 years. The operative corrective procedure consisted of a lower blepharoplasty and removal of the sheathed graft. Histology showed extended oil cysts and a surrounding capsulelike structure. The volume of the graft consisted mainly of oil cysts, and not healthy adipose tissue. The conclusion is that the harvesting procedure damages fragile adipocytes. The outcome of adipose tissue injection thus remains unpredictable. The only reliable guarantee for a good long term outcome after lipofilling is a specimen biopsy showing healthy adipose tissue. PMID- 11405368 TI - Ruptured pseudoaneurysm complicating an infected radial artery catheter: case report and review of the literature. AB - The authors present the 16th case of a pseudoaneurysm forming at the site of an infected radial artery catheter and only the third case that presented with rupture and hemorrhage requiring emergent operative repair. Radial artery catheters are quite safe, and most infections can be treated effectively with line removal and intravenous antibiotics. However, two factors correlate strongly with the subsequent development of pseudoaneurysms. Infection with Staphylococcus aureus was seen in 15 of 16 cases, and persistent signs of infection lasting more than 48 hours after the institution of antibiotic therapy and line removal were seen in 16 of 16 cases. Therefore, patients with S. aureus radial artery line infections with persistence of infection more than 48 hours after the induction of treatment are at high risk and should be observed closely for signs of pseudoaneurysm formation. Once a pseudoaneurysm has formed, surgical repair is required. Most recommend ligating the artery if there is pulsatile backbleeding from the distal stump and Allen's test shows good perfusion of the hand by the ulnar artery. PMID- 11405369 TI - Harvesting the abductor digiti minimi as a muscle plug with the lateral calcaneal artery skin flap. AB - The abductor digiti minimi muscle flap and the lateral calcaneal artery skin flap were raised as one combined flap and were used to cover plantar heel wounds with chronic osteomyelitis of the calcaneus. The combined flap was used successfully in 4 patients. The muscle component of the flap obliterated the dead space and provided a vascularized muscle over the debrided calcaneus, and the skin component acted as a sensate flap in the plantar heel. PMID- 11405370 TI - Providing patient protections. PMID- 11405371 TI - Reliability of secondary inferior pedicle (pyramidal) dermal technique in previously reduced gigantic breasts. PMID- 11405372 TI - Nasal tip metastasis of pharyngeal rhabdomyosarcoma. PMID- 11405373 TI - Accessory axillary breast tissue. PMID- 11405374 TI - Epidermoid carcinoma in cutaneous leishmaniasis scar. PMID- 11405375 TI - Surgical excision of cutaneous vascular lesions after percutaneous injection of n butyl 2-cyanoacrylate. PMID- 11405376 TI - Intraosseous zygomatic hemangioma. PMID- 11405377 TI - Modified tendon stripper for staged tendon grafting. PMID- 11405378 TI - Three thenar motor branches of the median nerve. PMID- 11405379 TI - Measurement of interincisal distance in the Indian population. PMID- 11405380 TI - A case of total nasal agenesis accompanied by Tessier no. 30 cleft. PMID- 11405381 TI - Palmoplantar keratoderma: usefulness of a surgical procedure. PMID- 11405382 TI - Xeroderma pigmentosum with a giant cutaneous horn. PMID- 11405383 TI - Specificity and total positive rate of head-up tilt testing potentiated with sublingual nitroglycerin in older patients with unexplained syncope. AB - The aim of this study was to assess the specificity and total positive rate of head-up tilt testing (HUTT) potentiated with sublingual nitroglycerin in detecting the vasovagal origin of unexplained syncope in the elderly, since the diagnostic value of this non-invasive test has not yet been proven in this age group. In a period of 3 years, 128 elderly patients (mean age 71.6+/-5.1 years, 50% males) with syncope of unknown origin, and 101 control subjects matched for age and gender were tilted upright to 60 degrees for 45 minutes. If syncope did not occur, sublingual nitroglycerin (0.4 mg) was administered, and observation was continued for 20 minutes. The positive response was defined as the reproduction of syncope or pre-syncope according to VASIS definition. During the unmedicated phase, syncope occurred in 26 patients (20.3%) and in no members of the control group. After nitroglycerin, 53 patients (41.4%) and 2 control subjects (2%) displayed syncope. The total positive rate of the test was 61.8% with a specificity of 98.0%. In conclusion, HUTT potentiated with sublingual nitroglycerin provides an adequate specificity and total positive rate in old patients with unexplained syncope; therefore it can be proposed as a useful diagnostic tool to detect the vasovagal origin of syncope not only in middle but also in advanced age. PMID- 11405384 TI - A multicenter, randomized, controlled trial to evaluate the safety profile, tolerability, and efficacy of rofecoxib in advanced elderly patients with osteoarthritis. AB - This 6-week study was conducted to test the efficacy, safety, and tolerability of rofecoxib (a selective COX-2 inhibitor) compared to nabumetone (a non-selective NSAID) and placebo in osteoarthritis (OA) patients aged 80 and older. Three hundred forty-one patients, mean age 83 years, were randomized. Allocations were made in an approximately 1:2:1:2 ratio (placebo: 12.5 mg rofecoxib: 25 mg rofecoxib: 1500 mg nabumetone). Least square mean changes from baseline in the primary efficacy endpoint, Patient Global Assessment of Disease Status, were as follows (with negative numbers indicating improvement): -14.85 mm for placebo; 25.34 mm for 12.5 mg rofecoxib; -25.40 mm for 25 mg of rofecoxib; and -25.95 mm for nabumetone (p<0.001 for all active treatments vs placebo.) Results from secondary endpoints, including the 3 WOMAC sub-scales (pain, stiffness, and disability) and the Investigator Global Assessment of Disease Status, were consistent with those for the primary endpoint. No significant between-group differences were observed in the proportions of patients who discontinued treatment due to either clinical or laboratory adverse experiences. Renal safety (edema and hypertension adverse experiences) was similar for rofecoxib and nabumetone. No gastroduodenal ulcers occurred; however, the demonstration of gastrointestinal risk with rofecoxib or nabumetone was beyond the scope of this trial. We conclude that in patients 80 years and older, rofecoxib, 12.5 mg and 25 mg once daily, demonstrated clinical efficacy for the treatment for OA as did 1500 mg of nabumetone. Rofecoxib and nabumetone were generally well tolerated in this elderly population. PMID- 11405385 TI - Effect of estrogen and progestin replacement on arterial stiffness indices in postmenopausal women. AB - Our objectives were to investigate whether long-term estrogen replacement therapy (ERT) is associated with a reduction in age-associated increases in arterial stiffness and blood pressure (BP), and whether the addition of progestin modifies the effects of estrogen. ERT has been found to have beneficial effects on cardiovascular risk. There are few data, however, delineating the effects of ERT on BP and arterial stiffness, and their age-associated changes. BP and aorto femoral pulse wave velocity (PWV) were measured in 134 postmenopausal volunteers, aged 51 to 90 years, from the Baltimore Longitudinal Study of Aging, screened to exclude clinical and occult cardiovascular disease, and classified as ERT non users (N=57) or ERT users (N=77). The latter group was further substratified according to the use of estrogen alone (N=32) or a combination of estrogen and progestins (N=45). ERT users showed similar body habitus, physical activity, and plasma lipids compared to non-ERT users. ERT was associated with an average 9.8 mmHg lower systolic BP (p<0.001), and a 6.3 mmHg lower pulse pressure (p<0.01) than in non-users. Multiple regression analysis showed that ERT was an independent predictor of lower SBP and PP (p<0.05). By analysis of covariance, ERT predicted a reduced age-associated increase in SBP, PP, and PWV (p<0.05). When systolic BP was >130 mmHg, the combination of ERT and progestins predicted a higher PWV than ERT alone. In conclusion, ERT in postmenopausal women can beneficially affect the vascular system, by reducing BP and the age-associated increase in arterial stiffness. The addition of progestins to ERT may reduce these beneficial effects. PMID- 11405386 TI - Age-associated changes in integral cholesterol and cholesterol sulfate concentrations in human scalp hair and finger nail clippings. AB - In contrast to surface lipids originating from the sebaceous glands, membrane forming integral lipids occur in keratinized tissues of skin, and skin appendages like fingernail plates or scalp hair. After removal of lipids of sebaceous origin by exhaustive solvent extraction, lyophilizing and hydrolyzing fingernail plate and scalp hair samples, fractions of integral cholesterol (CH) and cholesterol sulfate (CS) were quantified using gas chromatography. We studied these bound lipids and the serum lipids of 70 healthy subjects, aged 20.1 to 92.0 years. We observed higher amounts of CS in hair clippings of men than of women (775+/-241 vs 662+/-239 nmol/g hair, respectively). The highest amounts of CS were found in men with serum LDL-CH > 4.14 mmol/L; this subgroup also showed the highest CH values in fingernail clippings (2293+/-621 nmol/g nail). However, analysis of integral lipids of hair and fingernail plate clippings had little significance in detecting hypercholesterolemia in normal persons. An increase in integral CH levels in fingernail clippings with donor age was noted, independently of variations in serum CH or LDL-CH. This correlation proved to be significant in men (R=0.43), but not in women (R=0.38). In contrast, in women but not in men we found donor age correlated with internal CH of hair samples (R=0.43) and with CS of nail plates (R=-0.59), independently of serum CH or LDL-CH variations. This age-dependent decrease in CS levels might explain the previously observed higher incidence of brittle nails in women. Obviously, the metabolism of internal lipids CH and CS in fingernail and scalp hair differs between genders, and shows age associated changes. PMID- 11405387 TI - Age-associated diseases and conditions: implications for decreasing late life morbidity. AB - We discuss two types of age-associated diseases; aging-dependent such as Alzheimer's disease and congestive heart failure which increase logarithmically with age, versus age-dependent such as multiple sclerosis and amyotrophic lateral sclerosis which occur at proscribed ages, and then occurrence of new cases ceases or diminishes with further aging. Prevention strategies with both types emphasize postponement or delay of onset. The non-fatal aging-dependent diseases and conditions are an accumulating burden as we age, and increase overall morbidity in late years. These include Alzheimer's disease and other dementias, Parkinson's disease, loss of vision and hearing, incontinence, osteoporosis and hip fracture, osteoarthritis and depression. With mortality postponed, we will be living for many years at old and vulnerable ages. Life's quality will be reasonable for most. Still, increasing the chance that all will experience this desirable outcome requires pursuing the means to delay the onset of the physical and social events which we categorize as the non-fatal aging-dependent diseases and conditions. We must recognize that each added year occurs at the tip of an exponential curve where risk is maximal. PMID- 11405388 TI - Cognitive function and functional ability. A cross-sectional and longitudinal study at ages 85 and 95 in a non-demented population. AB - We studied the relationship between cognitive function and ability in daily life activities (ADL) at ages 85 (N=332) and 95 (N=63) cross-sectionally and longitudinally in representative populations of non-demented persons, in order to focus on manifestations related to healthy aging. Among the 85-year olds, 31% were independent, 42% were dependent on personal assistance in instrumental ADL (IADL) and 27% in personal ADL (PADL); the corresponding figures for the 95-year olds were 8, 40 and 52%, respectively. Regarding the relationship between cognitive function and ADL, the analysis could only be performed in the cross sectional data. At age 85 a significant trend was found regarding nearly all cognitive tests for both men and women, indicating that subjects who were ADL independent had better results than those dependent in IADL, and both IADL and PADL. At age 95, no such trend was found in males, but in females the trend was significant, and those dependent in ADL had poorer results in the separate cognitive tests. By combining the different cognitive tests into a compiled cognitive index, it was seen that the most ADL-dependent persons were more cognitively impaired both at ages 85 and 95, compared to persons who were ADL independent. Not only cognitive functioning (OR 3.3), but also mobility (OR 4.9) and tiredness (OR 3.3) were independent predictors for ADL-dependence among the 85-year olds. PMID- 11405389 TI - Outcome of medically unstable elderly patients admitted to a geriatric ward after hip fracture. AB - Outcome of surgical treatment is superior to that of conservative treatment for hip fractures. Nevertheless, for a number of patients, the operation is either delayed or unfit due to their unstable medical conditions. We retrospectively reviewed patients admitted to a geriatric ward after hip fracture, and investigated complications, functional outcome and survival in different cognitive, pre-fracture functional and treatment groups. Patients hospitalized (N=78) from January 1993 to June 1999 were included (1/2 demented, 1/3 fully dependent in Basic Activities of Daily Living, and 2/5 high operative risk patients). Following stabilization, 14 subjects (17. 9%) were operated. The mean and range of surgical delay was 9+/-7.2, and 3 to 30 days, respectively. Comparison between surgical and conservative treatment groups, and cognitive and pre-fracture functional groups showed no differences in age, gender, chronic medical conditions, fracture type, reasons for surgical delay or conservative approach, complications, survival curves and laboratory results. Thirteen operated patients were in ASA I + II grades, only 1 in ASA grades III + IV (low and high operative risk, American Society of Anesthesiologists grading system) (p=0.004). Functional outcome was similar in the surgical vs the conservative group, and intracapsular vs extracapsular fractures. ASA I + II patients had a higher survival rate compared to ASA III + IV patients (p=0. 02). We conclude that after stabilization of acute medical conditions, the most important preoperative consideration is the anesthetic risk, and surgical and conservative approaches may be equally considered in selected groups of elderly, frail patients with hip fracture who are medically unstable for more than a few days. PMID- 11405390 TI - Age-related changes in cytokine production by leukocytes in rhesus monkeys. AB - Using a variety of experimental rodent and human models, age-related alterations in cytokine production by immune cells have been described extensively. While the precise mechanism(s) responsible for such age-related changes in cytokine responses remain unclear, it seems likely that these changes may have a significant effect on immune cell function. In an attempt to clarify such changes in aging primates, we examined cytokine production by white cells derived from a controlled colony of rhesus monkeys (Macaca mulatta). Non-fractionated whole blood and peripheral blood mononuclear cells (PBMCs) were obtained from male monkeys of different ages (6-28 years), and were subsequently evaluated for their ability to express mRNA and protein for the cytokines, IL-10, IL-6, IFNgamma, IL 1beta, and TNFalpha, following in vitro stimulation with polyclonal mitogens. Our results suggest that white blood cells derived from aged rhesus monkeys exhibit a significant increase in their ability to produce the Th2-associated cytokine, IL 10, upon stimulation with lipopolysaccharide (LPS) when compared to white cells derived from younger counterparts. Similarly, a significant age-related decrease in the expression of the Th1-associated cytokine, IFNgamma, was also observed using phytohemagglutinin (PHA)-stimulated PBMCs. No significant age-related differences in the production of IL-1beta or TNFalpha were observed in response to any stimulation, but there was limited evidence of an age-related increase in IL-6 production. Overall, our results suggest that a possible systemic change from a Th0/Th1 to a Th2-like cytokine profile occurs in circulating leukocytes derived from aging primates. We believe that such age-related alterations in cytokine production may play a role in the reduced immune responses observed in elderly human populations. PMID- 11405391 TI - Correlates of knowledge of one's blood pressure and cholesterol levels among older members of a managed care plan. AB - We examined factors predicting knowledge of one's blood pressure, total cholesterol, and high-density lipoprotein levels (HDL) among older persons who reported a recent blood pressure or cholesterol test. Data come from a self administered, health risk assessment that was mailed to health plan members, age 55 and older, in a Santa Barbara, California medical group. Despite their universal access to health care and high levels of reported compliance with preventive health care screening practices, 41%, 49%, and 77% of respondents reported that they did not know their blood pressure, cholesterol, or HDL levels, respectively. After controlling for potential confounders, age and low income were inversely associated with the respondents' ability to report each level. In addition, current smoking and obesity were inversely associated with self reported knowledge of both cholesterol measures. Persons taking medication for hypertension or hypercholesterolemia were much more likely than those not taking medication to be able to report their blood pressure and cholesterol levels. Except for persons currently undergoing treatment for related conditions, these results suggest that a substantial proportion of the older persons at high risk for cardiovascular disease do not know their levels of these important biological risk factors. This lack of knowledge has important implications for public health education, and may hinder risk-reduction efforts among the elderly. PMID- 11405392 TI - Dissociation between coronary sinus and left atrial conduction in patients with atrial fibrillation and flutter. AB - INTRODUCTION: Coronary sinus (CS) recordings are routinely used during electrophysiologic studies for various supraventricular and ventricular arrhythmias with the understanding that they represent left atrial (LA) activity. However, the behavior of CS electrical activity during atrial arrhythmias has not drawn any special attention beyond standard considerations. METHODS AND RESULTS: The study population consisted of 9 patients (3 women; mean age 59 +/- 11 years) with atrial fibrillation (AF) and atrial flutter (AFL) who developed dissociation of conduction between the CS and posterior LA during spontaneous AF and AFL. In all patients, the LA and the CS were mapped using a 64-electrode basket catheter and a multipolar electrode catheter, respectively. The right atrium (RA) was mapped simultaneously using a 24-polar electrode catheter (7 patients) or a 64 electrode basket catheter (2 patients). Eight patients showed stable double potentials in CS recordings during AF (9 episodes) and AFL (3 episodes). During ongoing arrhythmias, the first row of potentials maintained a constant relationship with the RA activity, whereas the second row of potentials was discordant with the posterior wall of the LA in 7 patients and concordant in 2 patients. In 1 patient with counterclockwise AFL, CS activation was isolated from the posterior wall of the RA until it reached the distal portion of the CS, after which it entered the lateral region of the LA. In 1 patient, a macroreentrant LA tachycardia involving CS muscle was observed. Rapid atrial pacing from the proximal CS and extrastimuli produced longitudinal dissociation of CS activation in all patients. CONCLUSION: Conduction between the CS and posterior LA can be dissociated during spontaneous atrial arrhythmias and provocative proximal CS pacing. PMID- 11405393 TI - Is the coronary sinus really reflective of left atrial activation? PMID- 11405394 TI - Possible bradycardic mode of death and successful pacemaker treatment in a large family with features of long QT syndrome type 3 and Brugada syndrome. AB - INTRODUCTION: We recently identified a novel mutation of SCN5A (1795insD) in a large family with features of both long QT syndrome type 3 and the Brugada syndrome. The purpose of this study was to detail the clinical features and efficacy of pacemaker therapy in preventing sudden death in this family. METHODS AND RESULTS: The study group consisted of 116 adult family members: 60 carriers (29 males) and 56 noncarriers (28 males) of the mutant gene. Investigations included 24-hour Holter monitoring, ergometry, and electrophysiologic studies. Mean, lowest, and highest heart rate were lower in the carriers, but heart rate variability was comparable. In carriers, disproportional QT prolongation was present during bradycardia. No complex ventricular ectopy was recorded, and there were fewer isolated premature beats (both ventricular and atrial) in carriers. All patients were asymptomatic, except for two individuals who experienced syncope; in one of these patients, asystolic episodes (up to 9 sec) were repeatedly recorded. Prolonged HV intervals were present in 5 of 6 patients. Thirty carriers received a prophylactic backup pacemaker. During median follow-up of 4.5 years (range 0.0 to 22.6), their survival rate was 100%. There were five sudden deaths among the remaining 30 carriers without a pacemaker (P = 0.019). CONCLUSION: This family with a high incidence of nocturnal sudden death is characterized by bradycardia-dependent QT prolongation, intrinsic sinus node dysfunction, and generalized conduction abnormalities. There is a striking absence of complex ventricular ectopy, and pacemaker implantation was effective in preventing sudden death. These findings raise the possibility of a bradycardic rather than tachycardic mode of death. PMID- 11405395 TI - Ventricular tachycardia or conduction disease: what is the mechanism of death associated with SCN5A? PMID- 11405396 TI - Simultaneous analysis of heart rate variability and myocardial contractility during head-up tilt in patients with vasovagal syncope. AB - INTRODUCTION: The aim of this study was to evaluate simultaneously cardiac autonomic activity, through heart rate variability (HRV) analysis, and cardiac inotropic changes during head-up tilt (HUT) in patients with recurrent vasovagal syncope. METHODS AND RESULTS: Twelve subjects implanted with a permanent dual chamber pacemaker for recurrent vasovagal syncope characterized by marked bradycardia were studied. The tip of the right ventricular electrode was equipped with a sensor that measured peak endocardial acceleration (PEA) as an index of myocardial contractility. RR interval and PEA signals were acquired simultaneously and processed in the time and frequency (low frequencies [LF] and high frequencies [HF] of RR signal) domain during early HUT (T1), late HUT, or before syncope (T2). In the six subjects with positive HUT: (1) Abnormal heart rate oscillations were evidenced at T1 and discriminated this group from the negative group (LF/HF decreased by 46% from supine to T1, but increased by 55% in the negative group; P < 0.01 positive vs negative HUT). (2) Gradual diminution of the HF component was associated with an increase in PEA index during HUT with a correlation between PEA/RR interval (R = -0.8, P < 0.001), PEA/HF components (R = -0.6, P < 0.05). (3) Sympathetic stimulation responsible for changes in both HRV and PEA parameters occurred immediately before the faint (LF/LF+HF: 0.6 +/- 0.2 to 0.8 +/- 0.09; P < 0.05 T2 vs T1; PEA: 0.62 +/- 0.10G to 0.83 +/- 0.22G; P < 0.01 T2 vs T1). CONCLUSION: Our findings showed that a homogeneous subgroup of patients with recurrent vasovagal syncope and positive HUT exhibited abnormal cardiac autonomic and inotropic responses to an orthostatic stimulus. Continuous changes over time of HRV and PEA parameters highlight the dynamic behavior of the mechanisms leading to syncope. PMID- 11405397 TI - Prognostic significance of risk stratifiers of mortality, including T wave alternans, after acute myocardial infarction: results of a prospective follow-up study. AB - INTRODUCTION: Occurrence of sustained microvolt-level T wave alternans (TWA) at a specified heart rate has been suggested to predict life-threatening arrhythmic events, but its prognostic value has not been well established in patients who survived an acute myocardial infarction (AMI). The purpose of this prospective study was to assess the predictive significance of various noninvasive risk indicators of mortality, including TWA, in consecutive post-AMI patients with optimized medical therapy. METHODS AND RESULTS: In addition to a symptom-limited predischarge exercise test with measurement of TWA, mortality risk was assessed using heart rate variability, 24-hour ECG recordings, baroreflex sensitivity, signal-averaged ECG, QTc interval, QT dispersion, and echocardiographic wall motion index in 379 consecutive patients. Twenty-six patients (6.9%) died during a mean follow-up of 14 +/- 8 months. Sustained TWA was found in 56 patients (14.7%), none of whom died. Several risk variables, e.g., incomplete TWA test (inability to perform the exercise test or reach the required target heart rate of 105 beats/min), increased QRS duration on signal-averaged ECG, increased QT dispersion, long QTc interval, nondiagnostic baroreflex sensitivity result, and low wall-motion index, predicted all-cause mortality in univariate analyses. In multivariate analysis, the incomplete TWA test was the most significant predictor of cardiac death (relative risk 11.1, 95% confidence interval 2.4 to 50.8; P < 0.01). CONCLUSION: Sustained TWA during the predischarge exercise test after AMI does not indicate increased risk for mortality. An incomplete TWA test and several common risk variables provided prognostic information in this post-AMI population. PMID- 11405398 TI - Focal atrial tachycardia arising from the tricuspid annulus: electrophysiologic and electrocardiographic characteristics. AB - INTRODUCTION: Focal right atrial tachycardia (RAT) arising from the crista terminalis, para-Hisian, and coronary sinus os regions are well described. Less information exists regarding RAT arising from the nonseptal region of the tricuspid annulus (TA). METHODS AND RESULTS: From a consecutive series of 64 patients who had undergone successful radiofrequency ablation (RFA) of 67 RATs, the characteristics of 9 (13%) patients (6 men; mean age 50 +/- 20 years) with a TA focus were reviewed. The annular focus was localized to the inferoanterior TA in 7 and the superior TA in 2. Mean tachycardia cycle length was 371 +/- 66 msec. Mean activation time at the site of successful RFA in 9 of 9 patients was -43 +/- 11 msec. At 9.3 +/- 5.6 months of follow-up, 1 of 9 patients had recurrent tachycardia successfully treated with repeat RFA. In 7 of 9 patients with RAT from the inferoanterior TA, the surface ECG P wave morphology was upright in aVL, inverted in III and VI, and either inverted or biphasic with an initial negative deflection from V2 to V6. CONCLUSION: The TA is an important site of origin of RAT. In the present study, the inferoanterior region of the TA was a preferential site of origin with resulting characteristic P wave morphology. Knowledge of this anatomic distribution and P wave morphology allows targeted mapping and may facilitate successful RFA. PMID- 11405399 TI - On ridges, crests, and rings of fire: just location, or does the catheter point to the origins of focal tachycardia? PMID- 11405400 TI - Pace mapping of postinfarction scar to detect ventricular tachycardia exit sites and zones of slow conduction. AB - INTRODUCTION: The exit site and central common pathway of slow conduction are preferred sites to guide radiofrequency ablation of postinfarction ventricular tachycardia (VT). Both require inducibility of VT. In addition, their low amplitude hampers direct recording of potentials generated by activation in pathways of slow conduction. We hypothesized that pace mapping during sinus rhythm would help to detect the VT exit site and potentials generated by activation in pathways of slow activation. METHODS AND RESULTS: In 13 patients suffering from VT late after anterior (n = 10) or inferior (n = 3) myocardial infarction, stimulation was performed in scarred endocardium at 23.5 (range 13 to 36) sites per patient during arrhythmia surgery. Multielectrode recordings (64 sites) during stimulation at a fixed cycle length of 500 msec were obtained. Endocardial breakthrough sites distant (>2 cm) from the pacing site were found at 4.3 (range 3 to 19) pacing sites per patient. Low-amplitude discrete potentials (LADPs) could be detected between the pacing site and the breakthrough site in 2.3 (range 0 to 13) of 4.3 stimulation sequences. In these patients, 19 VTs were induced and the exit site determined. In 6 patients, the distant pacing breakthrough site was identical to the VT exit site; in 7 patients, no similar exit sites were found. LADPs during VT were found at a median 2.0 (range 0 to 14) sites per patient. CONCLUSION: Pace mapping of the postinfarction endocardial scar during sinus rhythm revealed 50% of the endocardial exit sites of VT and the same number of LADPs observed during VT. PMID- 11405401 TI - Atrioventricular reciprocating tachycardia involving twin atrioventricular nodes in patients with complex congenital heart disease. AB - INTRODUCTION: Histologic studies of autopsy specimens described the coexistence of two distinct AV nodes (so-called "Minckeberg sling" or "twin AV nodes") in specific congenital heart defects; however, the clinical electrophysiologic (EP) characteristics of twin AV nodes have not been characterized in detail. METHODS AND RESULTS: Since April 1993, a total of seven patients with complex congenital heart disease presented with AV reciprocating tachycardia suspected to be mediated by twin AV nodes. A common anatomic finding was AV discordance ([S,L,L] or [I,D,D]) with a malaligned complete AV canal defect in 5 of 7 patients. Intracardiac EP study was performed in five cases, and ablation was attempted in three patients with successful elimination of tachycardia inducibility by interruption or modification of 1 of the 2 AV nodes. Important EP characteristics included (1) the existence of two discrete nonpreexcited QRS morphologies, each with an associated His-bundle electrogram; (2) decremental as well as adenosine sensitive anterograde and retrograde conduction; and (3) inducible AV reciprocating tachycardia with anterograde conduction over one AV nodal pathway and retrograde conduction over the alternate AV nodal pathway. The existence of two AV nodes was further supported in the group treated with radiofrequency ablation by the development of transient accelerated junctional rhythm during energy delivery with an identical QRS morphology to that generated by anterograde conduction over the targeted AV node. CONCLUSION: Reciprocating tachycardia mediated by twin AV nodes can be a source of recurrent supraventricular tachycardia in patients with specific forms of complex congenital heart disease. Successful treatment with catheter ablation is possible. PMID- 11405402 TI - Brugada syndrome and supraventricular tachyarrhythmias: a novel association? AB - INTRODUCTION: The Brugada syndrome is a distinct form of idiopathic ventricular fibrillation characterized by a unique ECG pattern consisting of a right bundle branch block-like aspect and ST segment elevation in leads V1 to V3. As a high induction rate of ventricular tachyarrhythmias has been reported in Brugada syndrome, we hypothesized that this also may be true for supraventricular tachycardias in these patients. METHODS AND RESULTS: Between January 1995 and December 2000, we identified 35 consecutive patients with Brugada syndrome; 26 had a history of cardiac arrest or syncope and 9 were asymptomatic. All patients underwent electrophysiologic study, including an atrial and ventricular stimulation protocol. Ten patients (29%) were found to have supraventricular tachyarrhythmias (SVT) in addition to the Brugada syndrome. These 10 patients presented with aborted sudden cardiac death (n = 3) and/or a family history of sudden cardiac death (n = 4), syncope (n = 4), or primarily with a Brugada typical ECG, a positive family history, and palpitations (n = 2). Eight of them underwent genetic testing, but only 1 had a mutation in the SCN5A gene. In 6 patients, an AV nodal reentrant tachycardia was easily and reproducibly inducible. Two patients had clinical documented and inducible episodes of an atrial tachycardia (1 in addition to an AV nodal reentrant tachycardia). One patient had paroxysmal atrial fibrillation alternating with sinus rhythm, and 2 patients with accessory pathways were identified. CONCLUSION: This is the first description of an association of the Brugada syndrome with SVT. Thus, the arrhythmogenic substrate in Brugada syndrome may not be restricted to the ventricular level. Palpitations in this syndrome should raise the possibility of SVT. Conversely, in patients with SVT and aborted sudden cardiac death or syncope not related to SVT, the Brugada syndrome should be considered a possible additional electrophysiologic abnormality. PMID- 11405403 TI - Mechanisms for spontaneous changes in QRS morphology sometimes resembling torsades de pointes during reentrant ventricular tachycardia in a canine infarct model. AB - INTRODUCTION: Spontaneous changes in QRS morphology during sustained reentrant ventricular tachycardia, occurring gradually or abruptly, causing the tachycardia to be polymorphic, have been described in clinical cases. The purpose of this study was to determine the mechanism for such changes in a canine infarct model. METHODS AND RESULTS: Reentrant circuits were mapped in the epicardial border zone during sustained ventricular tachycardia in the canine heart, 4 days after left anterior descending coronary occlusion. In 10 tachycardias, there was either an abrupt change in QRS morphology or a gradual change that took up to 25 cycles. When the latter occurred, the ECG resembled torsades de pointes. Maps showed that the predominant mechanism for the change in QRS was a shift in the exit route by which the impulse left the reentrant circuit to activate the ventricles (9/10 tachycardias). Such shifts resulted from small changes in conduction velocity in segments of the circuit, either speeding or slowing, which modified the length of the functional lines of block. Movement of the circuit to a different region was responsible for the change in QRS in only one of these experiments, in which the reentrant mechanism also changed from functional to anatomic. CONCLUSION: Subtle changes in conduction in reentrant circuits can alter QRS morphology. Changes in the exit route from a stable reentrant circuit can cause the ECG characteristics to resemble torsades de pointes. PMID- 11405404 TI - Polymorphic ventricular tachycardia and torsades de pointes: beyond etymology. PMID- 11405405 TI - Role of wavelength adaptation in the initiation, maintenance, and pharmacologic suppression of reentry. AB - INTRODUCTION: The stability of reentry is thought to depend on a critical balance between the spatial extent of refractory tissue in a reentrant wave (i.e., wavelength lambda) and the reentrant path length. Because considerable evidence suggests that lambda changes continuously in space and time during abrupt rate changes associated with the onset of tachycardia, we hypothesized that beat-by beat adaptation of A to the dimensions of the reentrant path plays a central role in the mechanism of initiation of reentry. METHODS AND RESULTS: To investigate the dynamic relationship between lambda and path length during initiation of reentry, optical mapping with voltage-sensitive dyes was used in a guinea pig model of reentrant ventricular tachycardia (VT). In this model, a computer-guided laser obstacle precisely controlled the position and dimensions of the reentrant path. Under control perfusion and after addition of 15 microM d-sotalol, lambda was monitored during steady-state pacing, premature stimulation, and the initiating beats leading to nonsustained and sustained VT. During control perfusion, reentrant VT was reproducibly induced in 8 of 8 hearts, whereas in the presence of d-sotalol, reentry could only be initiated in 1 of 8 hearts due primarily to the failure of lambda to adapt to the reentrant path length. During successful initiation of VT, a consistent sequence was observed. The sequence was characterized by antidromic and orthodromic propagation around both sides of the anatomic obstacle, followed by unidirectional block of the antidromic impulse and persistence of reentry only if the A of the orthodromic impulse adapted to the reentrant path (lambda < path length). d-Sotalol prevented initiation of VT by altering lambda adaptation of the orthodromic wave; however, it failed to terminate ongoing VT because reverse use-dependence developed after several beats of tachycardia. CONCLUSION: In an experimental model where lambda, path length, and cellular action potentials were monitored during initiation of reentry, we found that, in contrast to termination, the initiation of reentry and the transition from nonsustained to sustained VT is strongly dependent on beat-to beat adaptation of lambda to the dimensions of the reentrant path. PMID- 11405406 TI - Excitable gap in canine fibrillating ventricular myocardium: effect of subacute and chronic myocardial infarction. AB - INTRODUCTION: The existence of an excitable gap during ventricular fibrillation (VF) has been suggested in several prior studies. However, the effects of myocardial infarction on the presence and duration of an excitable gap during VF have not been evaluated. METHODS AND RESULTS: Electrophysiologic study was performed in normal dogs and in dogs with subacute and chronic infarction. Experimental infarction was produced by left anterior descending coronary ligation. The excitable gap was determined indirectly using either evaluation of intrinsic wavefronts during VF or from the shortest activation interval at individual sites using recordings from a 112-electrode plaque sutured to the epicardial surface of the left ventricle. The excitable gap also was correlated to local electrophysiologic and anatomic properties. The excitable gap using the wavefront propagation method and shortest activation method was significantly longer in subacute infarction dogs (48 +/- 17 msec and 37 +/- 18 msec, respectively) and chronic infarction dogs (41 +/- 14 msec and 35 +/- 14 msec, respectively) than normal dogs (32 +/- 13 msec and 30 +/- 11 msec, respectively; P < 0.05 normal vs subacute and chronic infarction dogs in both methods). The excitable gap occupied approximately 30% and 27% of the VF cycle length in all three groups using the wavefront propagation and shortest activation method, respectively. The excitable gap correlated better with local ventricular refractoriness determined using the wavefront propagation method than with the shortest activation method, but not at all with refractoriness determined using extrastimulus testing. Tissue necrosis was noted in subacute infarction dogs and fibrosis in chronic infarction dogs, but the gap was not highly correlated with anatomic changes. CONCLUSION: During VF, an excitable gap exists in both normal and infarcted canine ventricular myocardium. It is significantly longer in the presence of infarction. These finding have implications for understanding the pathophysiology of VF and targeting antiarrhythmic therapies. PMID- 11405407 TI - Experimental and theoretical analysis of phase singularity dynamics in cardiac tissue. AB - INTRODUCTION: Quantitative analysis of complex self-excitatory wave patterns, such as cardiac fibrillation and other high-order reentry, requires the development of new tools for identifying and tracking the most important features of the activation, such as phase singularities. METHODS AND RESULTS: Image processing operations can be used to detect the phase singularity at the tip of a spiral wave. The phase space behavior of a spatiotemporal sequence of data may be reconstructed using time-series analysis. The phase singularities then are localized efficiently by computing the topologic charge density as the curl of the spatial phase gradient. We analyzed the singularity interaction dynamics of both experimentally observed and numerically simulated instances of quatrefoil reentry and found that the singularity behavior in the experimental preparations can be classified into three categories on the basis of how their separation changes with time. CONCLUSION: Topologic charge densities can be calculated easily and efficiently to reveal phase singularity behavior. However, the differences between theoretical and experimental observations of singularity separation distances indicate the need for more sophisticated numerical models. PMID- 11405408 TI - Use of tissue plasminogen activator in a stroke after radiofrequency ablation of a left-sided accessory pathway. AB - We describe the case of a 12-year-old girl who had a thromboembolic stroke after radiofrequency ablation of a left posterior accessory pathway involving a transseptal procedure. Symptoms of a stroke occurred 7 hours 15 minutes after completion of the procedure. Tissue plasminogen activator (tPA) was given 2 hours 30 minutes after the onset of symptoms, with complete resolution of her neurologic symptoms. No adverse effects from the tPA were seen. Because of the late onset of symptoms in this case, overnight in-hospital observation is warranted for patients who undergo radiofrequency ablation of a left-sided accessory pathway or an accessory pathway in a patient with the ability to shunt right to left. In this case, tPA was an effective and safe drug to use following a cerebral thromboembolic event occurring after a cardiac catheterization procedure. PMID- 11405409 TI - Myocardial ischemia: what factors determine arrhythmogenesis? AB - In ischemic myocardium, cellular architecture and heterogeneous changes in metabolic and ionic conditions interact to cause spatial and temporal heterogeneity of electrical properties predisposing to lethal ventricular arrhythmias. Yet, the details of their interaction, vis-a-vis the initiation and maintenance of reentry, remain poorly understood. Future studies are needed to address the mechanisms of the initial premature beat, reentry, and formation of wavebreaks contributing to ventricular fibrillation. New experimental methods including mathematical and transgenic mouse models are promising techniques to study these phenomena and thereby provide new insights into the fundamental mechanisms of ischemia-related arrhythmias. PMID- 11405410 TI - Recurrence of supraventricular tachycardia after slow pathway ablation for atrioventricular nodal reentrant tachycardia: what is the mechanism? PMID- 11405411 TI - Atrial pacing in hypertrophic cardiomyopathy with left mid-ventricular obstruction. PMID- 11405412 TI - Electrophysiologic effects of digoxin in the goat model of atrial fibrillation and its clinical implications. PMID- 11405413 TI - Relation of serum tetrachlorodibenzo-p-dioxin concentration to diet among veterans in the Air Force Health Study with background-level exposure. AB - To examine the contribution of various foods to exposure to 2,3,7,8-tetrachloro-p dibenzodioxin (TCDD) in a background-exposed U.S. population, serum TCDD levels were examined in relation to diet as assessed by a standard diet assessment instrument among men with no known unusual exposure to TCDD. Our subjects were male veterans aged 42-76 yr who were in the unexposed comparison group in the Air Force Health Study, a study of the health effects of exposure to Agent Orange and herbicides in Vietnam. Food consumption was assessed by a 126-item food frequency questionnaire. Two hundred and ninety veterans who had both TCDD levels and diet assessed in 1992 were included. In general, associations between serum TCDD and consumption of foods in specific groups or nutrients, that is, meats and fats, were not evident. Among younger men, fish and chicken intake were associated with higher serum TCDD levels. The results suggest that no single food group accounts for a large proportion of variation in TCDD exposure in older U.S. men. PMID- 11405414 TI - Inhibition of cholinesterase enzymes following a single dermal dose of chlorpyrifos and methyl parathion, alone and in combination, in pregnant rats. AB - Pregnant Sprague-Dawley rats (14-18 d of gestation) were treated with either a single dermal subclinical dose of 30 mg/kg (15% of dermal LD50) chlorpyrifos (O,O diethyl-O-[3,5,6-trichloro-2-pyridinyl] phosphorothioate) or a single dermal subclinical dose of 10 mg/kg (15% of dermal LD50) methyl parathion (O,O-dimethyl O-4-nitrophenyl phosphorothioate) or the two in combination. Chlorpyrifos inhibited maternal and fetal brain acetylcholinesterase (AChE) activity within 24 h of dosing, (48% and 67% of control activity, respectively). Following application of methyl parathion, peak inhibition of maternal and fetal brain AChE activity occurred at 48 h and 24 h after dosing (17% and 48% of control activity, respectively). A combination of chlorpyrifos and methyl parathion produced peak inhibition of maternal and fetal brain AChE activity at 24 h postdosing (35% and 73% of control activity, respectively). Maternal and fetal brain AChE activity recovered to various degrees of percentage of control 96 h after dosing. Application of methyl parathion or chlorpyrifos alone or in combination significantly inhibited maternal plasma butyrylcholinesterase (BuChE) activity. No significant inhibition of fetal plasma BuChE activity was detected. Peak inhibition of maternal liver BuChE occurred 24 h after application of methyl parathion or chlorpyrifos alone or in combination (64%, 80%, and 61% of control activity, respectively). Significant inhibition of placental AChE occurred within 24 h after application of methyl parathion or chlorpyrifos alone or in combination. The results suggest that methyl parathion and chlorpyrifos, alone or in combination, were rapidly distributed in maternal and fetal tissues, resulting in rapid inhibition of cholinesterase enzyme activities. The lower inhibitory effect of the combination could be due to competition between chlorpyrifos and methyl parathion for cytochrome P-450 enzymes, resulting in inhibition of the formation of the potent cholinesterase inhibitor oxon forms. The faster recovery of fetal plasma BuChE is attributed to the de novo synthesis of cholinesterase by fetal tissues compared to maternal tissues. PMID- 11405415 TI - Characterization of volatile organic compounds in smoke at experimental fires. AB - Significant associations between firefighting and cancer have been reported; however, studies finding toxic products of combustion at municipal fires have been limited by (1) technical difficulties encountered at the scene of working fires, (2) the lack of a coherent sampling strategy, and (3) the absence of verified sampling methods. The objective of the present study was to characterize the presence of volatile organic compound (VOC) combustion products in fire smoke. Air samples from experimental fires burning various materials commonly found at structural fires were collected into evacuated Summa canisters and analyzed for 144 target VOCs using cryogenic preconcentration and gas chromatography/mass spectroscopy (GC/MSD) methodology. The resulting chromatograms were characterized by a small number of predominant peaks, with 14 substances (propene, benzene, xylenes, 1-butene/2-methylpropene, toluene, propane, 1,2-butadiene, 2-methylbutane, ethylbenzene, naphthalene, styrene, cyclopentene, 1-methylcyclopentene, isopropylbenzene) being found in proportionately higher concentrations in all experimental fires and accounting for 65% (SD = +/-12%) by mass of total measured VOCs. Benzene, toluene, 1,3 butadiene, naphthalene, and styrene were found at higher concentrations than most other VOCs and increased with the time of combustion together with increasing levels of carbon monoxide. Benzene was found in the highest concentrations, with peak levels ranging from 0.6 ppm to 65 ppm, while the levels of 1,3-butadiene, styrene, and naphthalene peaked at 0.1, 0.4, and 3 ppm, respectively. This study revealed that there were no new or novel, toxic nonpolar VOCs resulting from the burning of common building materials. This is important in view of the studies that have found associations between firefighting and various forms of cancer. PMID- 11405416 TI - Investigation of the radioadaptive response in brain and liver of pUR288 lacZ transgenic mice. AB - The radioadaptive response, where a small priming dose of ionizing radiation can lessen the effects of subsequent exposure to a higher radiation challenge dose, was investigated in brain and liver within transgenic mice. Although it is well characterized in models in vitro, current radioadaptive response research has focused on particular cell types (i.e., lymphocytes) and does not provide comparative data for responses of multiple tissues within an organism. Transgenic animals are useful for such comparisons, because the transgene is integrated into all cells in the body. The pUR288 lacZ plasmid-based transgenic mouse model utilizes a plasmid vector allowing highly efficient recovery of mutational targets, including large size-change mutations that result from radiation exposure. Female C57BI/6 pUR288 lacZ mice were exposed to priming doses of 0.075- to 0.375-Gy x-rays over a 3-d period. After 3 wk, they received an acute challenge dose of 2.5-Gy x-rays. Spontaneous mutant frequencies in lacZ were significantly higher in liver than in brain (6.62 x 10(-5) vs. 3.51 x 10(-5)). In the absence of a priming dose, the 2.5-Gy challenge doubled the mutant frequency of both liver and brain (13.38 x 10(-5), and 7.63 x 10(-5) respectively). Priming doses of 0.15, 0.225, and 0.375 Gy significantly reduced (by 40%) the mutagenic effects of the 2.5-Gy challenge in brain. Restriction enzyme analysis of rescued mutant plasmids revealed a decrease in large size-change mutations at the three priming doses in brain. This study demonstrates the utility of this model for the investigation of radiological processes of large size-change mutations, as well as showing a radioadaptive response in brain, but not liver, of mice in vivo. PMID- 11405417 TI - Renal cortical mitochondrial dysfunction upon cadmium metallothionein administration to Sprague-Dawley rats. AB - A bolus dose of cadmium metallothionein (CdMT) produces renal proximal tubular dysfunction because it accumulates in the tubular epithelial cells and undergoes rapid degradation, releasing Cd. Morphologically, mitochondria appear to be the target organelle. The present study examined changes in renal cortical mitochondrial function following CdMT administration and investigated whether some of these effects could be ascribed to Cd2+ accumulation in the mitochondria. Sprague-Dawley rats were injected ip with 0.3 mg Cd as CdMT/kg and the animals were sacrificed after 6, 8, or 12 h. Two- to threefold increases in urinary protein excretion and LDH activity were evident at 8 h, with marked elevations (11- and 29-fold) thereafter. Renal cortical mitochondria were swollen and rounded at 12 h. The mitochondrial Cd level was 399 pmol/mg protein at 6 h and did not change significantly during the next 6 h; however, mitochondrial respiratory function declined with time. At 12 h, state 3 oxygen consumption, respiratory control ratio (RCR), and ADP:O (P/O) ratio were 48, 49, and 76% of control values, respectively, indicating inhibition of electron transfer and oxidative phosphorylation. The direct effect of Cd on mitochondrial function was examined by incubating mitochondria from untreated rats with 0.1-2 microM CdCl2. Rapid uptake of Cd resulted in concentration-dependent effects on respiration. After 1 min of incubation with 2 microM Cd, the mitochondria contained 262 microgCd/mg protein and state 3 respiration and RCR values were 75 and 33% of control levels, respectively. Thus, renal proximal tubular cell damage following a bolus dose of CdMT involves perturbations in mitochondrial respiration, brought on by the accumulation of Cd. PMID- 11405418 TI - Invariant object recognition in the visual system with error correction and temporal difference learning. AB - It has been proposed that invariant pattern recognition might be implemented using a learning rule that utilizes a trace of previous neural activity which, given the spatio-temporal continuity of the statistics of sensory input, is likely to be about the same object though with differing transforms in the short time scale. Recently, it has been demonstrated that a modified Hebbian rule which incorporates a trace of previous activity but no contribution from the current activity can offer substantially improved performance. In this paper we show how this rule can be related to error correction rules, and explore a number of error correction rules that can be applied to and can produce good invariant pattern recognition. An explicit relationship to temporal difference learning is then demonstrated, and from this further learning rules related to temporal difference learning are developed. This relationship to temporal difference learning allows us to begin to exploit established analyses of temporal difference learning to provide a theoretical framework for better understanding the operation and convergence properties of these learning rules, and more generally, of rules useful for learning invariant representations. The efficacy of these different rules for invariant object recognition is compared using VisNet, a hierarchical competitive network model of the operation of the visual system. PMID- 11405419 TI - Steady states in an iterative model for multiplicative spike-timing-dependent plasticity. AB - Recent experimental evidence suggests that synaptic plasticity depends on the precise timing of pre- and post-synaptic activity. In this paper, an iterative model for a multiplicative form of this spike-timing-dependent plasticity (mSTDP) is introduced. This model is incorporated into a neural network with many input cells coupled via excitation to a single output cell. Analysis of this network yields a criterion for the output cell to fire on every iteration, as well as general formulae for the steady-state output firing rate and the steady-state value to which all synaptic weights are driven by mSTDP. These characterize the basic state of network operation generated by mSTDP. PMID- 11405420 TI - Population density methods for large-scale modelling of neuronal networks with realistic synaptic kinetics: cutting the dimension down to size. AB - Population density methods provide promising time-saving alternatives to direct Monte Carlo simulations of neuronal network activity, in which one tracks the state of thousands of individual neurons and synapses. A population density method has been found to be roughly a hundred times faster than direct simulation for various test networks of integrate-and-fire model neurons with instantaneous excitatory and inhibitory post-synaptic conductances. In this method, neurons are grouped into large populations of similar neurons. For each population, one calculates the evolution of a probability density function (PDF) which describes the distribution of neurons over state space. The population firing rate is then given by the total flux of probability across the threshold voltage for firing an action potential. Extending the method beyond instantaneous synapses is necessary for obtaining accurate results, because synaptic kinetics play an important role in network dynamics. Embellishments incorporating more realistic synaptic kinetics for the underlying neuron model increase the dimension of the PDF, which was one-dimensional in the instantaneous synapse case. This increase in dimension causes a substantial increase in computation time to find the exact PDF, decreasing the computational speed advantage of the population density method over direct Monte Carlo simulation. We report here on a one-dimensional model of the PDF for neurons with arbitrary synaptic kinetics. The method is more accurate than the mean-field method in the steady state, where the mean-field approximation works best, and also under dynamic-stimulus conditions. The method is much faster than direct simulations. Limitations of the method are demonstrated, and possible improvements are discussed. PMID- 11405421 TI - Characterization of the variability of glutamatergic synaptic responses to presynaptic trains in rat hippocampal pyramidal neurons. AB - Excitatory postsynaptic currents from CA3 hippocampal neurons, elicited by trains of presynaptic action potentials either in mossy fibres or associative commissural fibres, have been analysed, by using a quantal analysis approach, in order to characterize their variability and the correlation among successive responses. As quantal parameters may change during the train according to the previous release events, correlation within consecutive EPSCs is expected. We tested simple hypotheses on how quantal parameters p and N may change on the basis of correlation detection in EPSCs. The statistical significance of these tests has been evaluated. The tests showed that, although simple binomial distributions can give a good description of synaptic responses at the level of single spikes, only stochastic chains can always account for correlations observed within the train. A systematic model fitting procedure has been developed and applied to extract information on the dynamics of synaptic transmission. As an application of this novel type of analysis, a measure of transmitted information to be associated with synaptic variability, a quantity that allows an estimate of the capability of the synapse to transmit reliable information in time, is proposed. We showed that this transmitted information depends on short-term plasticity and that the change in the type of short-term plasticity from facilitating to depressing obtained by increasing the extracellular calcium concentration results in a change of the related transmitted information. PMID- 11405422 TI - A simple white noise analysis of neuronal light responses. AB - A white noise technique is presented for estimating the response properties of spiking visual system neurons. The technique is simple, robust, efficient and well suited to simultaneous recordings from multiple neurons. It provides a complete and easily interpretable model of light responses even for neurons that display a common form of response nonlinearity that precludes classical linear systems analysis. A theoretical justification of the technique is presented that relies only on elementary linear algebra and statistics. Implementation is described with examples. The technique and the underlying model of neural responses are validated using recordings from retinal ganglion cells, and in principle are applicable to other neurons. Advantages and disadvantages of the technique relative to classical approaches are discussed. PMID- 11405423 TI - Precision of pulse-coupled networks of integrate-and-fire neurons. AB - Some sensory tasks in the nervous system require highly precise spike trains to be generated in the presence of intrinsic neuronal noise. Collective enhancement of precision (CEP) can occur when spike trains of many neurons are pooled together into a more precise population discharge. We study CEP in a network of N model neurons connected by recurrent excitation. Each neuron is driven by a periodic inhibitory spike train with independent jitter in the spike arrival time. The network discharge is characterized by sigmaW, the dispersion in the spike times within one cycle, and sigmaB, the jitter in the network-averaged spike time between cycles. In an uncoupled network sigmaB approximately = 1/square root(N) and sigmaW is independent of N. In a strongly coupled network sigmaB approximately = 1/square root(log N) and sigmaW is close to zero. At intermediate coupling strengths, sigmaW is reduced, while sigmaB remains close to its uncoupled value. The population discharge then has optimal biophysical properties compared with the uncoupled network. PMID- 11405424 TI - A model of the interaction between mood and memory. AB - This paper investigates a neural network model of the interaction between mood and memory. The model has two attractor networks that represent the inferior temporal cortex (IT), which stores representations of visual stimuli, and the amygdala, the activity of which reflects the mood state. The two attractor networks are coupled by forward and backward projections. The model is however generic, and is relevant to understanding the interaction between different pairs of modules in the brain, particularly, as is the case with moods and memories, when there are fewer states represented in one module than in the other. During learning, a large number of patterns are presented to the IT, each paired with one of two mood states represented in the amygdala. The recurrent connections within each module, the forward connections from the memory module to the amygdala, and the backward connections from the amygdala to the memory module, are associatively modified. It is shown how the mood state in the amygdala can influence which memory patterns are recalled in the memory module. Further, it is shown that if there is an existing mood state in the amygdala, it can be difficult to change it even when a retrieval cue is presented to the memory module that is associated with a different mood state. It is also shown that the backprojections from the amygdala to the memory module must be relatively weak if memory retrieval in the memory module is not to be disrupted. The results are relevant to understanding the interaction between structures important in mood and emotion (such as the amygdala and orbitofrontal cortex) and other brain areas involved in storing objects and faces (such as the inferior temporal visual cortex) and memories (such as the hippocampus). PMID- 11405425 TI - Long-term care and a good quality of life: bringing them closer together. AB - Long-term care policies and programs in the United States suffer from a major flaw: They are balanced toward a model of nursing home care that, regardless of its technical quality, tends to be associated with a poor quality of life for consumers. This article proposes quality-of-life domains-namely, security, comfort, meaningful activity, relationships, enjoyment, dignity, autonomy, privacy, individuality, spiritual well-being, and functional competence. It argues that these kinds of quality-of-life outcomes are minimized in current quality assessment and given credence only after health and safety outcomes are considered. Five trends are reviewed that might lead to a more consumer-centered emphasis on quality of life: the disability rights movement, the emphasis on consumer direction, the growth of assisted living, increasing attention to physical environments, and efforts to bring about culture change in nursing homes. Building on these trends, the article concludes with strategies to move beyond current stalemates and polarized arguments toward forms of long-term care that are more compatible with a good quality of life. PMID- 11405426 TI - Community long-term care services: what works and what doesn't? PMID- 11405427 TI - Asking the right questions: the key to discovering what works in home care. PMID- 11405428 TI - What works? Maine's statewide uniform assessment and home care planning system tells all. PMID- 11405429 TI - Improving communication between researchers and policy makers in long-term care: or, researchers are from Mars; policy makers are from Venus. PMID- 11405430 TI - The Positively Aging teaching materials improve middle school students' images of older people. AB - PURPOSE: The Positively Aging program is an innovative set of interdisciplinary teaching materials that uses examples from geriatrics and gerontology to teach sixth through eighth grade curricular elements. The purpose of this study was to determine if use of the Positively Aging teaching materials by regular classroom teachers could change middle school students' images of elders. DESIGN AND METHODS: At the beginning of the 1998-1999 school year, students at two San Antonio, Texas, middle schools were asked to draw a typical older person. These drawings were coded as positive, neutral, or negative portrayals of elders. One school then used the Positively Aging materials as part of the curriculum; the other school served as the control. Second drawings were obtained from the students at the end of the school year and compared to those from baseline. RESULTS: Both drawings were completed by 60% of students at the intervention school and 55% of students at the control school. Of the 782 paired drawings from the intervention school, 34% were more positive at Time 2 compared to 25% of 591 paired drawings from the control school (chi2 = 13.9, p < .001). In addition, only 20% of the second drawings from the intervention school were more negative than the first drawing compared to 27% from the control school (chi2 = 11.3, p < .001). Using a generalized logit model, we adjusted for each student's baseline drawing (positive-neutral-negative), grade level, gender, ethnic group, and socioeconomic status. After adjustment, students in the intervention school were more likely to draw positive (odds ratio [OR] 1.48, 95% confidence interval [CI] 1.13, 1.94) or positive and neutral images (OR 1.58, 95% CI 1.21, 2.19) at follow up compared to the control school. IMPLICATIONS: This controlled study demonstrated that use of the Positively Aging teaching materials and activities moved middle school students toward a more positive view of elders. Interdisciplinary teaching materials based on geriatrics and gerontology can be successfully developed and tested in public school systems to affect attitudes about aging. PMID- 11405431 TI - Racial, ethnic, and cultural differences in the dementia caregiving experience: recent findings. AB - PURPOSE: This research reviewed studies that compare two or more racial, ethnic, national, or cultural groups on aspects of the dementia caregiving experience. DESIGN AND METHODS: Electronic databases were searched to find studies published between 1996 and 2000 in peer-reviewed journals that met the above criteria. RESULTS: Twenty-one studies based on 18 samples were identified. These articles included comparisons involving the following groups of caregivers: African Americans, Chinese, Chinese Americans, Koreans, Korean Americans, Latinos, Whites, and residents of 14 European Union countries. Consistent with previous research, White caregivers were more likely to be spouses when compared to other groups. White caregivers tended to report greater depression and appraised caregiving as more stressful than African American caregivers. Findings were mixed regarding differences in coping and social support, but suggested that minority groups may not have more available support than Whites. Common methodological limitations were a lack of noncaregiving control groups and failure to test specific pathways by which the grouping variable (e.g., race) exerts its impact on outcome variables. IMPLICATIONS: Future studies in this area should use both quantitative and qualitative research methods to specify the pathways by which race, ethnicity, and culture affect the caregiving experience, and should expand their focus beyond the primary caregiver to include the effects of caregiving on families and networks. PMID- 11405432 TI - Mismanaging prescription medications among rural elders: the effects of socioeconomic status, health status, and medication profile indicators. AB - PURPOSE: This study assessed the extent to which community-dwelling rural older adults mismanage their prescription medication regimens and predicted mismanagement of medications from selected socioeconomic, health status, and medication profile characteristics. DESIGN AND METHODS: Personal interviews with 499 community-dwelling adults aged 66 and over taking at least one prescription medication and living in a rural region of the Southeast. With approximately equal numbers of African American and white men and women, the SUDAAN multiple logistic regression procedure was used to predict the mismanagement of prescription medications. RESULTS: The mismanagement of prescribed medication regimens is relatively common among older adults. Those more likely than others to mismanage their regimens are African American, younger, in poorer mental health, with more acute care physician visits, and those who find payment for their medications to be problematic. IMPLICATIONS: The implications of the findings for what is known about the self-modification of drug regimens, targeting prescription drug cost benefits or interventions, and the limitations of the study are discussed. PMID- 11405433 TI - Toward a threshold for subthreshold depression: an analysis of correlates of depression by severity of symptoms using data from an elderly community sample. AB - PURPOSE: The prevalence of depressive symptoms in elderly adults is high, yet the criteria to identify clinically significant depression may leave many elders undiagnosed and untreated. We explored the demographic and risk factor profiles of two groups, one with more severe depression and one with less severe depression. DESIGN AND METHODS: The data come from the Duke University Established Populations for Epidemiologic Studies of the Elderly (EPESE) baseline survey of 4,162 community-dwelling adults aged 65 or older. RESULTS: The prevalence of depression meeting criteria of the Center for Epidemiologic Studies Depression scale (CES-D) and sub-threshold depression was 9.1% and 9.9%, respectively. In ordinal logistic regression, both CES-D and subthreshold depression were associated with impairment in physical functioning, disability days, poorer self-rated health, use of psychotropic medications, perceived low social support, female gender, and being unmarried. IMPLICATIONS: Depression appears to exist along a continuum, with demographic and social and physical health predictors of subthreshold depression similar to predictors of depression as defined by the CES-D scale. PMID- 11405434 TI - Paranoid ideation among elderly African American persons. AB - PURPOSE: The prevalence and correlates of paranoid ideation were investigated. DESIGN AND METHODS: On the basis of a sample of 998 independently living elderly African American persons in a cross-sectional study, the study used the Brief Symptom Inventory to measure paranoid ideation and 14 independent variables, including demographic characteristics, cognitive deficit, depression, self reported memory functioning, emotional and instrumental support, stressful life events, limitation of daily activities, self-rated health status, and self-rated hearing and vision. RESULTS AND IMPLICATIONS: Paranoid ideation (symptoms of paranoia) was found in 10% of this sample. A multiple regression analysis of the data revealed that of the 14 independent variables used in this study, 6 (income, instrumental support, hearing, stressful life events, self-reported memory deficit, and depression) showed a significant relationship with paranoid ideation. PMID- 11405435 TI - Are persons with cognitive impairment able to state consistent choices? AB - PURPOSE: This study examined the decision-making capacity of persons with cognitive impairment with respect to their everyday care preferences and choices. This is the first in a series of articles to report on findings from a larger study that examines choice, decision making, values, preferences, and practices in everyday care for community-dwelling persons with cognitive impairment and their family caregivers. DESIGN AND METHODS: Fifty-one respondent pairs, or dyads, were interviewed, that is, persons with cognitive impairment (n = 51) and their family caregivers (n = 51). All persons with cognitive impairment were interviewed twice within a week using a parallel interview to determine stability and accuracy of responses. The family caregiver was interviewed once. RESULTS: Persons with mild to moderate cognitive impairment (i.e., Mini-Mental State Exam scores 13-26) are able to respond consistently to questions about preferences, choices, and their own involvement in decisions about daily living, and to provide accurate and reliable responses to questions about demographics. IMPLICATIONS: Including the perspective of persons with cognitive impairment in both research and practice has the potential to enhance their autonomy and improve their quality of life. PMID- 11405436 TI - State legislation concerning individuals with dementia: an evaluation of three theoretical models of policy formation. AB - PURPOSE: This study examined the formation of state government policies concerning persons with dementia. In particular, we identified variables associated with the passage of pertinent state laws by testing three theoretical models of policy formation: the iron triangle, the policy system, and an integrated model. METHODS: We sampled 44 states and counted the number of laws concerning individuals with dementia that passed during the 1998 legislative sessions. We constructed nine independent variables to represent the three theoretical models of state policy formation. Then the number of legislative actions were regressed onto each model. RESULTS: The integrated model that consisted of political actors, legislative features, and environmental inputs met goodness-of-fit criteria for a maximum likelihood regression analysis (chi2/df = 1.55). The advocacy effort of the Alzheimer's Association, number of legislative champions, supply of special care beds for persons with dementia, and recent policy activity were related significantly with increased legislative activity concerning individuals with dementia. IMPLICATIONS: Policy responses concerning persons with dementia increasingly have been created within state governments, and we identified variables associated with the passage of pertinent state laws. This research also contributed to the advancement of comparative state policy research by contrasting the three theoretical models of policy formation. PMID- 11405437 TI - Preparing future aging advocates: the Oklahoma Aging Advocacy Leadership Academy. AB - PURPOSE: This article describes the organization, operation, and evaluation of the Oklahoma Aging Advocacy Leadership Academy (OAALA). DESIGN AND METHODS: The Academy was designed to assure the continuation of future cohorts of advocates and volunteers for aging programs and services. It is a statewide training program consisting of 10 weekend sessions. The curriculum incorporates age related content and advocacy/leadership/voluntary skills training. Evaluation consisted of: (a) a pre-post assessment of perceived familiarity with advocacy and volunteer leadership issues; (b) post-Academy participant assessments of the quality of various aspects of the Academy; (c) post-Academy written statements about areas that were valuable and in need of change; and (d) a compilation of student volunteer and advocacy activities. RESULTS: Evaluation results indicate that the OAALA led to significant increases in perceived familiarity with issues and very positive participant assessments of quality. In addition, many participants spontaneously initiated advocacy and volunteer activities while enrolled in the Academy. IMPLICATIONS: Our experience with the Academy suggests that it is a feasible means of preparing individuals to become successful old-age advocates and volunteer leaders. PMID- 11405438 TI - Measurement sensitivity and the Minimum Data Set depression quality indicator. AB - PURPOSE: The purpose of this study was to determine the accuracy of the prevalence rating of depression in nursing homes as flagged on the Minimum Data Set (MDS) quality indicator report. DESIGN AND METHODS: Research Staff measured depression symptoms and compared the results with the prevalence of disturbed mood symptoms documented by nursing home (NH) staff on the MDS in two samples of residents living in different NHs. The homes had been flagged on the nationally mandated MDS quality indicator report as having unusually low (Site 1) or high (Site 2) prevalence rates of depression. RESULTS: The percentages of residents determined by research staff interview assessments to have probable depression in the two resident samples were not significantly different (49% vs. 55%, respectively) between homes. The staff in the home flagged on the MDS quality indicator report as having a high depression prevalence rate identified significantly more residents who also had scores indicative of probable depression on the resident interviews for follow-up mood assessments than did the home with a low quality indicator prevalence rate (78% vs. 25%, respectively). IMPLICATIONS: The prevalence of the depression quality indicator may be more reflective of measurement processes than of depression outcomes. Factors that may affect the difference in detection rates are discussed. PMID- 11405439 TI - In-home assessment of dementia by nurses: experience using the CERAD evaluations. Consortium to Establish a Registry for Alzheimer's Disease. AB - PURPOSE OF STUDY: To maximize respondent participation in a study of the prevalence, incidence, and natural history of dementia. DESIGN AND METHODS: Clinical research nurses were trained to carry out evaluations for dementia in the home. We describe the assessment and training procedures used and note the advantages and drawbacks of this approach. RESULTS: Nurse identification of the presence of dementia agreed well with that of medical specialists (kappa = 0.84), but was slightly poorer regarding type of dementia (kappa = 0.71). IMPLICATIONS: Use of nurses for such activity need not be limited to epidemiological studies, but is relevant also in clinical practice. PMID- 11405440 TI - Response to "Ageism in gerontological language". PMID- 11405441 TI - A review of the effects of multiple stressors on aquatic organisms and analysis of uncertainty factors for use in risk assessment. AB - Risk assessment procedures use toxicity tests in which organisms are subjected to chemicals under otherwise constant and favorable experimental conditions. Because variable and suboptimal environmental conditions are common aspects of natural ecosystems, the hazard of underestimation of risk arises. Therefore, an uncertainty factor is used in the extrapolation of results of standard toxicity tests to field situations. The choice for these uncertainty factors is based on little ecological evidence. This review discusses studies on the toxicity of various chemicals to aquatic organisms, modified by temperature, nutritional state and salinity, excluding papers on changes in bioavailability of compounds. Collected data were analyzed quantitatively to evaluate the validity of toxicity data obtained from standard toxicity tests in the laboratory under field conditions. Generally, organisms living under conditions close to their environmental tolerance limits appeared to be more vulnerable to additional chemical stress. Usually, increasing temperature and decreasing food or nutrient level raised toxicity. The influence of salinity was less clear; metal toxicity increased with decreasing salinity, toxicity of organophosphate insecticides increased with higher salinity, while for other chemicals no clear relationship between toxicity and salinity was observed. The interactions can be explained by several physical and physiological processes, acting on factors such as bioavailability, toxicokinetics, and sensitivity of organisms. Quantitative analysis of data indicated that an uncertainty factor for the laboratory to field extrapolation should be smaller than one for an ecosystem in a temperate region, while a factor greater than one would be appropriate for systems nearby discharge points of cooling water. The factor should be greater than one when varying nutritional state is concerned, but smaller than one with respect to salinity. Dependent on the effect parameter used, the differences in toxicity between laboratory and relevant field situations ranged from a factor of 2.6 to 130 and 1.7 to 15 for the two temperature conditions and 1.2 to 10 for nutritional state. A salinity increase from freshwater to marine water decreased toxicity by a factor of 2.1. However, as less extreme salinity changes are more relevant under field conditions, the change in toxicity is probably much smaller. To obtain uncertainty factors that sufficiently protect natural systems without being overprotective, additional research is required. PMID- 11405442 TI - A review of quantitative studies of benzene metabolism. AB - Benzene is a ubiquitous, highly flammable, colorless liquid that is a known hematotoxin, myelotoxin, and human leukemogen. Benzene-induced toxicity in animals is clearly mediated by its metabolism. The mechanisms of acute hemato- and myelotoxicity in humans are almost certainly the same as in animals, and there is compelling evidence that metabolism is requisite for the induction of leukemia in humans. A very large number of experimental investigations of benzene metabolism have been conducted with animals, both in vivo and in vitro. There have also been many investigations of benzene metabolism in humans and with human tissues, Although the blood or tissue concentrations of benzene metabolites in humans resulting from benzene exposure have never been measured. Further, a number of mathematical models of benzene metabolism and dosimetry have been developed. In this article, we consider results from both experimental and mathematical modeling research, with particular emphasis on the last decade, and discuss the factors that are likely to be most influential in the metabolism of benzene. PMID- 11405443 TI - Health risks associated with inhaled nasal toxicants. AB - Health risks of inhaled nasal toxicants were reviewed with emphasis on chemically induced nasal lesions in humans, sensory irritation, olfactory and trigeminal nerve toxicity, nasal immunopathology and carcinogenesis, nasal responses to chemical mixtures, in vitro models, and nasal dosimetry- and metabolism-based extrapolation of nasal data in animals to humans. Conspicuous findings in humans are the effects of outdoor air pollution on the nasal mucosa, and tobacco smoking as a risk factor for sinonasal squamous cell carcinoma. Objective methods in humans to discriminate between sensory irritation and olfactory stimulation and between adaptation and habituation have been introduced successfully, providing more relevant information than sensory irritation studies in animals. Against the background of chemoperception as a dominant window of the brain on the outside world, nasal neurotoxicology is rapidly developing, focusing on olfactory and trigeminal nerve toxicity. Better insight in the processes underlying neurogenic inflammation may increase our knowledge of the causes of the various chemical sensitivity syndromes. Nasal immunotoxicology is extremely complex, which is mainly due to the pivotal role of nasal lymphoid tissue in the defense of the middle ear, eye, and oral cavity against antigenic substances, and the important function of the nasal passages in brain drainage in rats. The crucial role of tissue damage and reactive epithelial hyperproliferation in nasal carcinogenesis has become overwhelmingly clear as demonstrated by the recently developed biologically based model for predicting formaldehyde nasal cancer risk in humans. The evidence of carcinogenicity of inhaled complex mixtures in experimental animals is very limited, while there is ample evidence that occupational exposure to mixtures such as wood, leather, or textile dust or chromium- and nickel containing materials is associated with increased risk of nasal cancer. It is remarkable that these mixtures are aerosols, suggesting that their "particulate nature" may be a major factor in their potential to induce nasal cancer. Studies in rats have been conducted with defined mixtures of nasal irritants such as aldehydes, using a model for competitive agonism to predict the outcome of such mixed exposures. When exposure levels in a mixture of nasal cytotoxicants were equal to or below the "No-Observed-Adverse-Effect-Levels" (NOAELs) of the individual chemicals, neither additivity nor potentiation was found, indicating that the NOAEL of the "most risky chemical" in the mixture would also be the NOAEL of the mixture. In vitro models are increasingly being used to study mechanisms of nasal toxicity. However, considering the complexity of the nasal cavity and the many factors that contribute to nasal toxicity, it is unlikely that in vitro experiments ever will be substitutes for in vivo inhalation studies. It is widely recognized that a strategic approach should be available for the interpretation of nasal effects in experimental animals with regard to potential human health risk. Mapping of nasal lesions combined with airflow driven dosimetry and knowledge about local metabolism is a solid basis for extrapolation of animal data to humans. However, more research is needed to better understand factors that determine the susceptibility of human and animal tissues to nasal toxicants, in particular nasal carcinogens. PMID- 11405444 TI - MR grading of temporomandibular joint fluid: association with disk displacement categories, condyle marrow abnormalities and pain. AB - The purpose of this study was to investigate temporomandibular joint (TMJ) effusion on magnetic resonance (MR) images, and its association with specific categories of disk displacement, bone marrow abnormalities and pain. From a series of 523 consecutive TMJ MR imaging studies of patients referred to imaging because of pain and dysfunction, those with TMJ effusion, defined as an amount of fluid that exceeded the maximum amount seen in a control group of asymptomatic volunteers, were analysed. The selected patients were reassessed and the amount of TMJ fluid was graded bilaterally according to a set of reference films. Other parameters recorded included disk displacement categories and condyle marrow abnormalities. Pain self-records were obtained from the patients immediately before MR imaging. The association between the recorded parameters and TMJ pain was analysed with t-tests and regression analysis. Of the 523 patients, 70 (13%) had TMJ effusion, which was unilateral in 61%. Only 9% of the 70 patients had effusion bilaterally, whereas bilateral disk displacement was found in 80%. In the 76 joints with effusion, 83% showed two specific categories of disk displacement at closed mouth. Condyle marrow abnormalities were found in 31% of the 70 patients, mostly on one side, and in 24% of the 76 joints. An in-patient regression analysis of the side difference in TMJ pain showed that effusion and condyle marrow abnormalities were significant pain-increasing factors. In conclusion, patients with TMJ effusion represent a subgroup with pain and dysfunction with more severe intra-articular pathology than those with disk displacement but no other joint abnormalities. PMID- 11405445 TI - MR evidence of temporomandibular joint fluid and condyle marrow alterations: occurrence in asymptomatic volunteers and symptomatic patients. AB - The purpose of this study was to estimate the frequency and amount of temporomandibular joint (TMJ) fluid, as well as the frequency and type of condyle marrow alterations in asymptomatic volunteers and compare to patients with TMJ pain and dysfunction. Proton-density and T2 weighted magnetic resonance (MR) images of the TMJs of 62 asymptomatic volunteers and 58 symptomatic patients were analysed for fluid and condyle marrow alterations as well as disk position. The amount of fluid (increased T2 signal) was characterized as none, minimal, moderate or marked and related to the disk position. The differentiation between moderate and marked fluid was based on the maximum amount of fluid seen in the volunteers; more than this amount was categorized as marked fluid. The marrow of the mandibular condyle was categorized as normal, edema (increased T2 signal) or sclerosis (decreased proton-density and T2 signal) and related to fluid and disk position. In the 62 asymptomatic volunteers, 50 (81%) had none or minimal and 12 (19%) had moderate TMJ fluid. In the 58 symptomatic patients, 40 (69%) had none or minimal and 18 (31%) had moderate or marked fluid. Both in volunteers and patients, moderate fluid could be seen in joints with normal disk position, but was significantly associated with disk displacement. In the 62 volunteers, no signal abnormalities in the condyle marrow were found. In the 58 patients, six (10%) had abnormal bone marrow. These six patients had disk displacement and two had moderate or marked fluid. Marked fluid and condyle marrow abnormalities were therefore not encountered in any of the asymptomatic volunteers but in about 10% of the patients. PMID- 11405446 TI - Relationship between temporomandibular joint pain and magnetic resonance imaging findings of internal derangement. AB - In terms of clinical decision-making in instances of temporomandibular disorders (TMD) and orofacial pain, there is controversy in the literature over the diagnostic significance of the temporomandibular joint (TMJ)-related variable disk-condyle relationship (DCR). The purpose of this study was to investigate whether in patients with TMJ-related pain, the variable of TMJ pain may be linked to magnetic resonance (MR) imaging findings of internal derangement (ID). The study comprised 163 consecutive TMJ pain patients. Criteria for including a patient were report of orofacial pain referred to the TMJ, and the presence of uni- or bilateral TMJ pain during palpation, during function, and/or during unassisted or assisted mandibular opening. Bilateral sagittal and coronal MR images were obtained to establish the prevalence of TMJ ID types. Analysis of the data revealed the presence of TMJ pain to be associated with significantly more MR imaging diagnoses of ID than an absence of ID (P<0.001), and disk displacement without reduction than disk displacement with reduction (P<0.001). Using chi square analysis, the results showed a significant relationship between the presence of TMJ-related pain and the MR imaging diagnosis of TMJ ID (P=0.001), and TMJ ID type (P=0.000). Use of the Kappa statistical test indicated poor diagnostic agreement between the presence of TMJ pain and the MR imaging diagnosis of ID (K=0.16). The results suggest that the clinical variable of TMJ pain may have a significant effect on the prevalences of MR imaging diagnoses of TMJ ID. The data confirm the biological concept of DCR as a diagnostic approach in patients with signs and symptoms of TMJ-related pain. PMID- 11405447 TI - Nasal airway changes after Le Fort I--impaction and advancement: anatomical and functional findings. AB - Twenty patients were studied prospectively to assess intranasal anatomical changes and functional changes resulting from a one-piece Le Fort I-osteotomy with anterior and superior positioning of the maxilla. Presurgical and 3 months postsurgical rhinological inspection, anterior rhinomanometry and acoustic rhinometry were performed. Interalar width was measured and cephalograms were used to assess maxillary movement. Rhinoscopically, three septal perforations (15%) were noticed. Turbinate enlargement was less common postoperatively. Interalar width increased significantly. These findings correlate with a significant increase in cross-sectional diameter at the Isthmus nasi revealed by acoustic rhinometry 3 months postoperatively. The mean total nasal airflow measured by anterior rhinomanometry was unchanged indicating no increase in resistance despite decreased intranasal dimensions in cases where the impaction is not higher than 5 mm. PMID- 11405448 TI - Mandibular invasion of lower gingival carcinoma in the molar region: its clinical implications on the surgical management. AB - The spread pattern of a tumour and its extent in the mandible are important in the management of gingival cancer. Sixteen patients with gingival squamous cell carcinoma (SCC) involving the mandible in the molar region were included in this study. Resection specimens of the mandibular bone and adjacent cancer were histologically analysed to identify the type and characteristics of invasion and were compared with the radiological features. Our results showed that the actual width of invasion was underestimated to a greater extent than the actual depth of invasion. For horizontal aspects, four dentate cases had horizontal intramedullary spread underneath intact mucosa or cortical bone extended from the main foci of tumour that infiltrated through the occlusal surface. For vertical aspects, nerve invasion took place in only one of 16 specimens, while five cases showed downward infiltration beyond the inferior alveolar canal without nerve involvement, so that the pattern of tumour spread was mostly transmedullary rather than perineural in previously non-irradiated cases. These cases with deep infiltration showed the infiltrative type of invasion in the dentate mandible. And when the tumour was related with previous dental extractions or curettage, it tended to be more extensive than what was predicted from an imaging point of view. These pathological and clinical features affecting the tumour spread should be considered in the management of gingival SCC in the molar region. PMID- 11405449 TI - The usefulness of intraoral ultrasonography in the evaluation of oral cancer. AB - Many studies focused on the tumour thickness in oral squamous cell carcinomas, suggesting a relationship with the occurrence of cervical metastasis. Accurate preoperative assessment of the tumour thickness of oral cancer would provide useful information for targeting those patients who need elective treatment of the neck. Some useful diagnostic aids to evaluate oral cancer are computed tomography (CT), magnetic resonance imaging (MRI), and intraoral ultrasonography. The purpose of the present study is to compare intraoral ultrasonography with CT and MRI in delineating the disease extent and in measuring the tumour thickness of oral carcinoma. Thirty-nine patients with oral cancer were preoperatively evaluated with intraoral ultrasonography, and CT, and in 26 of them MRI was carried out. High-quality ultrasonographic images were obtained and the tumour thickness was measured within 1 mm. However, in most tumours less than 5.0 mm in thickness, CT and MRI could not detect a sufficient density difference from the normal tissue to accurately delineate the extent of the tumour. There was a significant correlation between measurements by intraoral ultrasonography and the histological sections. The present study shows that ultrasonography is superior to CT and MRI in assessment of the primary lesion of oral carcinoma. PMID- 11405450 TI - Influence of chemotherapy on endosteal implant survival and success in oral cancer patients. AB - Little is known about the effect of chemotherapy on the osseointegration and survival of endosteal dental implants. In a retrospective study, two groups of patients were compared: one group consisting of 30 oral cancer patients received postsurgical adjuvant chemotherapy with either cis- or carboplatin and 5 fluorouracil in three cycles and were treated subsequently with 106 dental implants placed in the mandible; the other group consisting of 17 patents suffering from oral cancer was prescribed with 54 dental implants placed in the mandible after oncological surgery. No patient was treated with radiotherapy. Twenty patients in the first group were successfully provided with a prosthetic superstructure (mean time of function: 35.8 months) compared to 16 patients in the second group (mean time of function: 36.2 months). The observation time was 10 years. A life-table analysis based on defined success parameters demonstrated no significant difference between implant survival in either group. It was concluded that chemotherapy with cis- or carboplatin and 5-fluorouracil was not detrimental to the survival and success of dental implants in the mandible. PMID- 11405451 TI - Recombinant human erythropoietin in the treatment of head and neck tumour anaemia. AB - At the time of first diagnosis, patients with squamous cell carcinoma in the head and neck are often in the advanced stage of their disease, therefore surgery is not a viable option for treatment. These patients also present frequently a high grade of anaemia as a result of either the malignant process itself or of the following therapy. The incidence of anaemia and the need for transfusion depends on several factors, such as the type and intensity of radiotherapy and radiochemotherapy. Multimode therapeutic concepts such as radio-chemotherapy are being applied with increasing frequency, resulting in an ever increasing need for transfusion with great effects on the patient's quality of life. Even more important to tumour patients is the role of the haemaglobin (Hb) value as a prognostic factor for survival and/or local tumour control. A large number of studies show that recombinant human erythropoietin (r-HuEPO) is effective in the treatment of tumour-induced anaemia and prevention and correction of chemotherapy and radiotherapy-induced anaemia. The simultaneous application of r-HuEPO with chemotherapy can prevent patients with head and neck tumours from developing anaemia or can reduce the extent of the anaemia and the need for transfusion. Comparable effects were observed both in patients undergoing platinum-based and non-platinum-based chemotherapy. The direct correlation between anaemia, tumour hypoxia and poor response to radio and/or chemotherapy has been clinically proven. Recombinant human erythropoietin administration improves the therapeutic outcome and the patients' prognosis. PMID- 11405452 TI - Lethal invasive mucormycosis: case report and recommendations for treatment. AB - A case of lethal invasive mucormycosis (IM), a rare fungal infection which predominantly affects immunocompromised patients, is reported in a 73-year-old female patient who presented with a cervical abscess. The patient had asthma treated with steroids and had previously undiagnosed diabetes mellitus. Despite surgical treatment and parenteral antibiotic therapy, there was fatal progression of the condition. The pathogenesis, histological appearances and treatment of mucormycosis are discussed, particularly the importance of urgent histological examination of debrided tissue to distinguish this condition from necrotizing fasciitis (NF) earlier than microbiological culture alone would allow, thus permitting the early introduction of appropriate antifungal therapy. PMID- 11405453 TI - A comparative study on four screw-plate locking systems in sheep: a clinical and radiological study. AB - Four mandibular screw-plate locking systems were studied in sheep. Three to four centimetres angular continuity resections of the mandible were performed and the defects bridged with pre-bent angular reconstruction plates fixed with four screws in the body part and three in the ramus. Each type was used four times. Clinical and radiological examinations were carried out 1 and 2 months later. All sheep were able to eat and ruminate normally throughout the postoperative period. Radiology revealed that 6/16 plates and 5/112 screws fractured during the follow up period. In one type, no fracture occurred. Screw fracture was confined to just one reconstruction system. Six of 16 mandibles showed slight to moderate bone resorption under the plate. The results point to the internal shortcomings of the systems tested. PMID- 11405454 TI - Initial experiences using a new implant based distraction system for alveolar ridge augmentation. AB - With this case report we demonstrate a new technique useful in augmenting alveolar ridges after resection procedures or atrophic loss of volume. The augmentation is done by means of a new osteogenic distractor (manufactured by ACE Surgical Supply Co., Inc., Brockton, MA, USA), the distraction procedure is according to Ilizarov's principles. PMID- 11405455 TI - Primary orbital liposarcoma: case report and review of the literature. AB - Primary orbital liposarcoma is an extraordinarily rare tumour. Lack of specific diagnostic findings, scarcity of published series and a primary anatomical location make its management complicated. Surgery should conserve visual function in well located, low-grade tumours that do not invade ocular structures. Complementary RT should also be performed. PMID- 11405456 TI - Unusually large cystic adenomatoid odontogenic tumour of the maxilla: case report. AB - Adenomatoid odontogenic tumour (AOT) of the maxillary sinus is extremely rare. A case of a gigantic AOT that occupied the maxillary sinus is described. PMID- 11405457 TI - A multifocal neurinoma of the hypoglossal nerve with motor paralysis confirmed by electromyography. AB - A rare case of neurinoma in a 72 year-old Japanese woman derived from the hypoglossal nerve is reported. The tumour was composed of three interconnected nodules occurring simultaneously in the left submandibular and sublingual regions. The lesion, which presented as a neck mass, caused a slight left-sided hemiparesis of the tongue with tongue deviation to the affected side not noticed by the patient. An electromyographic (EMG) study revealed decreased muscle activity on the left side of the tongue muscle, indicating dysfunction of the hypoglossal nerve. EMG was useful for diagnosis. PMID- 11405458 TI - Craniofacial distraction osteogenesis: a review of the literature: Part 1: clinical studies. AB - A review of the literature dealing with distraction osteogenesis (DO) of the craniofacial skeleton, provided by a PUBMED search (National Library of Medicine, NCBI; revised 3 April 2000) from 1966 to December 1999 was conducted. Key words used in the search were distraction, lengthening, mandible, mandibular, maxilla, maxillary, midface, midfacial, monobloc, cranial, craniofacial and maxillofacial. This search revealed 285 articles. One hundred and nine articles were clinically orientated and were analysed in detail in this study. The type of distraction, indications, age, type of surgery, distraction rates and rhythms, latency and contention periods, amount of lengthening, follow-up period, relapse, complications and the nature of the distraction device were analysed. This review revealed that 828 patients underwent DO of the craniofacial skeleton; 579 underwent mandibular DO, 129 maxillary DO, 24 simultaneous mandibular and maxillary DO and 96 midfacial and/or cranial DO. Craniofacial DO has proven to be a major advance for the treatment of numerous congenital and acquired craniofacial deformities. Treatment protocols and success criteria for craniofacial DO are suggested on the basis of these results. There is still, however, a lack of sufficient data, especially on follow-up and relapse, so that treatment strategies have to be validated by long-term studies in the future. PMID- 11405459 TI - Cellulose crystallites. AB - This article discusses advances in understanding the structural and physicochemical characteristics of suspensions of cellulose crystallites prepared by acid hydrolysis of natural cellulose fibres. Consideration of recent developments in visualization of crystallite ultrastructure may provide clues to suspension behavior. In addition, novel applications in a diverse range of fields are presented, from iridescent pigments to biomolecular NMR studies. PMID- 11405460 TI - Matrix isolation and spectroscopic characterization of perfluorinated ortho- and meta-benzyne. AB - The matrix isolation and spectroscopic characterization of two C6F4 isomers, the perfluorinated o-benzyne 4 and the m-benzyne 5, is reported. UV photolysis of tetrafluorophthalic anhydride 6 in solid argon at 10 K results in the formation of CO, CO2, and 1,2-didehydro-3,4,5,6-tetrafluorobenzene (4) in a clean reaction. On subsequent 350 nm irradiation 4 is carbonylated to give the cyclopropenone 7. 1,3-Didehydro-2,4,5,6-tetrafluorobenzene (5) was synthesized by UV irradiation of 1,3-diiodo-2,4,5,6-tetrafluorobenzene (8) via 2,3,4,6-tetrafluoro-5 iodophenylradical 9. Photolysis of 8 in solid neon at 3 K produces good yields of both radical 9 and benzyne 5, while in argon at 10 K no reaction is observed. Thus, the photochemistry in neon at extremely low temperature markedly differs from the photochemistry in argon. The formation of 5 from 8 via 9 is reversible, and annealing the neon matrix at 8 K leads back to the starting material 8. The benzynes 4 and 5 and the radical 9 were characterized by comparison of their matrix IR spectra with density functional theory (DFT) calculations. PMID- 11405461 TI - The synthesis of (+)-allopumiliotoxin 323B'. AB - This paper describes the synthesis of (+)-allopumiliotoxin 323B' (1) using the intramolecular [3 + 2]-cycloaddition reaction of the (Z)-N-alkenylnitrone 4. This synthesis began with (R)-tert-butyl-3-hydroxy-pent-4-enoate [(R)-13] which was obtained by enzymatic resolution with Amano PS lipase. A series of manipulations gave intermediate 17 and in situ coupling with 4-benzoyloxybutanal lead to the (Z)-N-alkenylnitrone 4 which underwent an intramolecular [3 + 2]-cycloaddition reaction to give the isoxazolidine 3 as the major cycloadduct. Isoxazolidine 3 provided the piperidinone 24 which upon diastereofacial selective addition of MeMgBr gave the required tertiary alcohol 25. Formation of the indolizidine core 2 was achieved by an intramolecular S(N)2 reaction. The side chain was assembled from a Wittig reaction between the phosphorane 8 and the enantiomerically pure aldehyde 9. Further modifications afforded the aldehyde 7 which underwent an aldol condensation with the potassium enolate of the indolizidone core 2. Dehydration gave the enone 37 which was converted into the anti-diol 38 by intramolecular hydride reduction. Finally, deprotection of the BOM protecting group gave (+)-allopumiliotoxin 323B' (1). PMID- 11405462 TI - Trapping reactions of an intermediate containing a tungsten-phosphorus triple bond with alkynes. AB - The thermolysis of the phosphinidene complex [Cp*P[W(CO)5]2] (1) in toluene in the presence of tBuC(triple bond)CMe leads to the four-membered ring complexes [[[eta2-C(Me)C(tBu)]Cp*(CO)W(mu3-P)[W(CO)3]][eta4:eta1:eta1 P[W(CO)5]WCp*(CO)C(Me)C(tBu)]] (4) as the major product and [[W[Cp*(CO)2]W(CO)2WCp*(CO)[eta1:eta1-C(Me)C(tBu)]](mu,eta3:eta2:eta1-P2[W(CO)5]] (5). The reaction of 1 with PhC(triple bond)CPh leads to [[W(Co)2[eta2 C(Ph)C(Ph)]][(eta4:eta1-P(W(CO)5]W[Cp*(CO)2)C(Ph)C(Ph)]] (6). The products 4 and 6 can be regarded as the formal cycloaddition products of the phosphido complex intermediate [Cp*(CO)2W(triple bond)P --> W(CO)5] (B), formed by Cp* migration within the phosphinidene complex 1. Furthermore, the reaction of 1 with PhC(triple bond)CPh gives the minor product [[[eta2:eta1 C(Ph)C(Ph)]2[W(CO)4]2][mu,eta1:eta1-P[C(Me)[C(Me)]3C(Me)][C(Ph)](C(Ph)]] (7) as a result of a 1,3-dipolaric cycloaddition of the alkyne into a phosphaallylic subunit of the Cp*P moiety of 1. Compounds 4-7 have been characterized by means of their spectroscopic data as well as by single-crystal X-ray structure analysis. PMID- 11405463 TI - Photosensitization of crystalline and amorphous titanium dioxide by platinum(IV) chloride surface complexes. AB - Anatase, rutile, and amorphous titania powders were surface-modified by grinding with PtCl4 and H2[PtCl6]. Only the anatase modification afforded hybrid photocatalysts capable of degradation of 4-chlorophenol (4-CP) with visible light, with sufficient stability towards decomplexation. Grinding with K2[PtCl4] produced materials of only low photocatalytic activity. Most efficient photocatalysts contained up to 2 wt% of PtIV. At higher surface loading the excess fraction of the complex is desorbed into the aqueous solution. Scavenging experiments with benzoic acid and tetranitromethane revealed that hydroxyl radicals are produced by the primary reduction of oxygen by conduction band electrons generated through electron injection from a postulated surface platinum(III) complex. It is proposed that the latter is formed from a charge transfer ligand-to-metal (CTLM) excited state through homolysis of the Pt-Cl bond. Accordingly. the primary oxidation of 4-CP may occur by adsorbed chlorine atoms, the intermediary existence of which was demonstrated by scavenging experiments with phenol. PMID- 11405464 TI - Derivatization of glucose and 2-deoxyglucose for transition metal complexation: substitution reactions with organometallic 99mTc and Re precursors and fundamental NMR investigations. AB - Synthetic strategies for the bifunctionalization of glucose and 2-deoxyglucose at position C-1 for transition metal coordination are reported. In particular organometallic technetium and rhenium complexes for potential use in diagnostic nuclear medicine were synthesized and investigated. Specifically, a common iminodiacetic acid (IDA) moiety was O-glycosidically connected through an ethylene spacer group to produce the pure alpha- (in case of 2-deoxyglucose) and beta-anomer (in case of glucose). Reaction of the sugar derivatives with the organometallic precursor [M(H2O)3(CO)3]+ (M = 99mTc, Re) produced single products in high yield, which are water-soluble and water-stable. The displacement of the three water molecules of the metal precursor and thus the tridentate coordination of the metal-tricarbonyl core exclusively via the amine and the two carboxylic acid functionalities of the IDA chelate was verified by means of 1D and 2D 1H NMR spectroscopy, mass spectrometry, and IR spectroscopy. The radioactive-labeled products (99mTc) proved their excellent stability in vitro in physiological phosphate buffer (pH = 7.4) and human plasma over a period of 24 h at 37 degrees C. PMID- 11405465 TI - Synthesis, reactivity, and structure of strictly homologous 18 and 19 valence electron iron nitrosyl complexes. AB - The 18 and 19 valence electron (VE) nitrosyl complexes [Fe(NO)('pyS4')]BF4 ([1]BF4) and [Fe(NO)('pyS4')] (2) have been synthesized from [Fe('pyS4')]x ('pyS4'(2-) = 2,6-bis(2-mercaptophenylthiomethyl)pyridine(2-)) and either NOBF4 or NO gas. Complex [1]BF4 was also obtained from [Fe(CO)('pyS4')] and NOBF4. The cation [1]+ is reversibly reduced to give 2. Oxidation of 2 by [Cp2Fe]PF6 afforded [Fe(NO)('pyS4')]PF6 ([1]PF6). The molecular structures of [1]PF6 and 2 were determined by X-ray crystallography. They demonstrate that addition of one electron to [1]+ causes a significant elongation of the Fe-donor atom bonds and a bending of the FeNO angle. Density functional calculations show that the unpaired electron in 2 occupies an orbital, which is antibonding with respect to all Fe ligand interactions. As expected from qualitative Molecular Orbital (MO) theory, it has a large contribution from a pi* type NO orbital. The nu(NO) frequency decreases from 1893 cm(-1) in [1]BF4 to 1648 cm(-1) in 2 (in KBr). The antibonding character of the unpaired electron explains the ready reaction of 2 with excess NO to give [Fe(NO)2('pyS4')] (3), the facile NO/CO exchange of 2 to afford [Fe(CO)('pyS4')], and the easy oxidation of 2 to [1]+. PMID- 11405466 TI - Calcium oxalate crystals in tomato and tobacco plants: morphology and in vitro interactions of crystal-associated macromolecules. AB - Plants form calcium oxalate crystals with unique morphologies under well controlled conditions. We studied the morphology of single calcium oxalate monohydrate (whewellite) crystals extracted from tomato and tobacco leaves. These crystals have a pseudotetrahedral shape. We identified the (101), (101) or (102), (110), and (hk0) faces as stable faces. The morphology is chiral with unique handedness. We also show that calcium oxalate monohydrate crystals isolated from tomato, tobacco, and bougainvillea leaves contain macromolecules rich in Gly, Glx, and Ser. Crystal-associated macromolecules extracted from tomato and tobacco influence the morphology of calcium oxalate monohydrate crystals grown in vitro, promoting preferential development of the [120] faces. Furthermore, crystal associated macromolecules from tobacco promote nucleation of calcium oxalate monohydrate crystals, whereas model polypeptides do not have any significant effect on nucleation. These results imply an active role of the crystal associated macromolecules in the formation of pseudotetrahedral shapes in vitro, and these properties may in part be responsible for the unique chiral morphology of the natural pyramidal-shaped crystals. PMID- 11405467 TI - Identification of sequence selective receptors for peptides with a carboxylic acid terminus. AB - Split-and-mix libraries of resin-bound "tweezer" receptors have been prepared and screened to identify receptors for dye-labelled tripeptides. The receptors incorporate a diamidopyridine unit to serve as a specific recognition site for the CO2H group, leading to strong and selective receptors for peptide guests with a CO2H terminus. The role of the dye-label, attached to the peptide guest to allow visualisation of selective recognition events in the screening experiments, has also been examined and was found to have a significant influence on the binding selectivities. PMID- 11405468 TI - Formation of ternary complexes by coordination of (diethylenetriamine) platinum(II) to N1 or N7 of the adenine moiety of the antiviral nucleotide analogue 9. AB - The synthesis of (Dien)Pt(PMEA-N1), where Dien = diethylenetriamine and PMEA2- = dianion of 9-[2-(phosphonomethoxy)ethyl]adenine, is described. The acidity constants of the threefold protonated H3[(Dien)Pt(PMEA-N1)]3+ complex were determined and in part estimated (UV spectrophotometry and potentiometric pH titration): The release of the proton from the (N7)H+ site in H4[(Dien)Pt(PMEA N1)]3+ occurs with a rather low pKa (= 0.52+/-0.10). The release of the proton from the -P(O)2(OH) group (pKa = 6.69+/-0.03) in H[(Dien)Pt(PMEA-N1)]+ is only slightly affected by the N1-coordinated (Dien)Pt2+ unit. Comparison with the acidic properties of the H[(Dien)Pt(PMEA-N7)]+ species provides evidence that in the (Dien)Pt(PMEA-N7) complex in aqueous solution an intramolecular, outer-sphere macrochelate is formed through hydrogen bonds between the -PO3(2-) residue of PMEA2- and a PtII-coordinated (Dien)NH2 group; its formation degree amounts to about 40%. The stability constants of the M[(Dien)Pt(PMEA-N1)]2+ complexes with M2+ = Mg2+, Ca2+, Ni2+, Cu2+ and Zn2+ were measured by potentiometric pH titrations in aqueous solution at 25 degrees C and I = 0.1 M (NaNO3). Application of previously determined straight-line plots of log K(M(R-PO3))M versus pK(H(R PO3)H for simple phosph(on)ate ligands. R-PO3(2-), where R represents a non inhibiting residue without an affinity for metal ions, proves that the primary binding site of (Dien)Pt(PMEA-N1) is the phosphonate group with all metal ions studied; in fact, Mg2+, Ca2+ and Ni2+ coordinate (within the error limits) only to this site. For the Cu[(Dien)Pt(PMEA-N1)]2+ and Zn[(Dien)Pt(PMEA-N1)]2- systems also the formation of five-membered chelates involving the ether oxygen of the CH2-O-CH2-PO3(2-) residue could be detected; the formation degrees are about 60% and 30%, respectively. The metal-ion-binding properties of the isomeric (Dien)Pt(PMEA-N7) species studied previously differ in so far that the resulting M[(Dien)Pt(PMEA-N7)]2+ complexes are somewhat less stable, but again Cu2+ and Zn2+ also form with this ligand comparable amounts of the mentioned five-membered chelates. In contrast, both M[(Dien)Pt(PMEA-N1/N7)]2+ complexes differ from the parent M(PMEA) complexes considerably; in the latter instance the formation of the five-membered chelates is of significance for all divalent metal ions studied. The observation that divalent metal-ion binding to the phosphonate group of (Dien)Pt(PMEA-N1) and (Dien)Pt(PMEA-N7) is only moderately inhibited (about 0.2-0.4 log units) by the twofold positively charged (Dien)Pt2+ unit at the adenine residue allows the general conclusion, considering that PMEA is a nucleotide analogue, that this is also true for nucleotides and that consequently participation of, for example, two metal ions in an enzymatic process involving nucleotides is not seriously hampered by charge repulsion. PMID- 11405469 TI - About the stereoelectronics of the intramolecular addition of allylsilanes to aldehydes. AB - (Z)-omega-Trimethylsilyl-(omega-2)-alken-1-ols are readily accessible by consecutive superbase metalation and silylation of (omega-1)-alken-1-ols. These versatile intermediates may be oxidized to give the corresponding (Z)-omega trimethylsilyl-(omega-2)-alkenals which, in the presence of trifluoroacetic acid, can be converted into 2-vinylcycloalkanols such as 2-vinylcyclohexanol (2), isopulegol (4), and bis(2-vinylcyclobutyl) ether (8). The stereochemical outcome of these cyclization reactions suggests the interference of a novel electrodynamic effect. PMID- 11405470 TI - Structure and thermoelectric properties of the new quaternary bismuth selenides A(1-x)M(4-x)Bi(11+x)Se21 (A = K and Rb and Cs; M = Sn and Pb)--members of the grand homologous series Km(M6Se8)m(M(5+n)Se(9+n)). AB - Several members of the new family A(1-x)M(4-x)Bi(11+x)Se21 (A = K, Rb, Cs; M = Sn, Pb) were prepared by direct combination of A2Se, Bi2Se3, Sn (or Pb), and Se at 800 degrees C. The single-crystal structures of K(0.54)Sn(3.54)Bi(11.46)Se21, K(1.46)Pb(3.08)Bi(11.46)Se21, Rb(0.69)Pb(3.69)Bi(11.31)Se21, and Cs(0.65)Pb(3.65)Bi(11.35)Se21 were determined. The compounds A(1-x)M(4-x)Bi(11+x) Se21 crystallize in a new structure type with the monoclinic space group C2/m, in which building units of the Bi2Te3 and NaCl structure type join to give rise to a novel kind of three-dimensional anionic framework with alkali-ion-filled tunnels. The building units are assembled from distorted, edge-sharing (Bi,Sn)Se6 octahedra. Bi and Sn/Pb atoms are disordered over the metal sites of the chalcogenide network, while the alkali site is not fully occupied. A grand homologous series Km(M6Se8)m(M(5+n)Se(9+n)) has been identified of which the compounds A(1-x)M(4-x)Bi(11+x)Se21 are members. We discuss here the crystal structure, charge-transport properties, and very low thermal conductivity of A(1 x)M(4-x)Bi(11+x)Se21. PMID- 11405471 TI - Theoretical characterization of photoisomerization channels of dimethylpyridines on the singlet and triplet potential energy surfaces. AB - Photoexcitations and photoisomerizations due to low-lying n pi* and pi pi* excited states of dimethylpyridines are investigated by density functional theory, CASSCF, CASPT2 and MRCI methodologies. Mechanistic details for the formation of Dewar dimethylpyridines and the interconversions of dimethylpyridines are rationalized through the characterization of minima and transition states on the singlet and triplet potential energy surfaces of relevant intermediates. Our present theoretical schemes suggest that Mobius dimethylpyridine intermediate 14 and azabenzvalene intermediate 10 can serve as possible precursors to Dewar dimethylpyridines and singlet phototransposition products, respectively. The calculations suggest that an S1(pi pi*)/S0 conical intersection in dimethylpyridines 2 is involved in the formation of 14. An azabenzvalene 10 might be formed through S2(pi pi*)/S1(n pi*) interaction followed by an S1/S0 decay in dimethylpyridine 6. Calculated barriers of isomerizations from 14 to Dewar dimethylpyridine 7 and from 10 to 4 are 8.4 and 28.5 kcal mol(-1) at the B3LYP/6-311 G** level, respectively. In the suggested triplet multistage transposition mechanism, an out-of-plane distorted geometry 19 due to vibrational relaxation of the T1(3B1) excited state of 3,5 dimethylpyridine 6 is a precursor of the interconversion of 6 to 2.4 dimethylpyridine 4. The formation of a triplet azaprefulvene 21 with a barrier of 20.7 kcal mol(-1) is a key step during the triplet migration process leading to another out-of-plane distorted structure 27. Subsequent rearomatization of 27 completes the interconversion of 6 with 4. Present calculations provide some insight into the photochemistry of dimethylpyridines at 254 nm. PMID- 11405472 TI - Solvated CH5+ in liquid superacid. AB - The transition states for methane activation in liquid superacid have been studied by experimentally determined secondary kinetic deuterium isotope effects (SKIEs) and computational chemistry. For the first time, the SKIEs on hydrogen/deuterium exchange of methane have been measured by using the methane isotopologues in homogeneous liquid superacid (2HF/SbF5). To achieve high accuracy of the SKIEs, the rate constants for pairs of methane isotopologues were simultaneously measured in the same superacid solution by using NMR spectroscopy. Density functional theory (DFT) and high-level ab initio methods have been employed to model possible intermediates and transition states, assuming that the superacids involved in the exchange reactions are H2F+ ions solvated by HF. Only the unsolvated superacid H2F+ is found to be strong enough to protonate methane, yielding the methonium ion solvated by HF as a potential energy minimum. In contrast, the (HF)x-solvated H2F+ superacids (x = 1-4) do not appear to be strong enough to yield stable solvated methonium ions. However, such ions show up as parts of the transition states of the exchange in which the methonium ions are solvated by (HF)x. The calculated DFT activation barrier is in good agreement with that experimentally observed. PMID- 11405473 TI - Resorcarenes in the boat conformation as building blocks for hydrogen-bonded assemblies including two ammonium cations. AB - Crystal structures are reported for various co-crystals of rccc-resorcarenes with triethylammonium chloride. Usually, two molecules of a C2v-symmetric tetraester 2 in the boat conformation are linked through four hydrogen-bonded chloride anions to give dimeric assemblies. Two of the chloride anions may be replaced by four hydrogen-bonded ethanol molecules in an otherwise similar structure. These assemblies, which consist of six or eight components, posses voluminous, negatively charged chambers in which two triethylammionium cations, 3+, are included as guests by strong electrostatic and hydrogen-bonding interactions. The host-guest N-H...Cl hydrogen bonds were clearly detected at 173 K. These are the first examples of hydrogen-bonded, solid-state capsules trapping two ions of the same charge in close proximity. In the 1:2 complex with 3+ Cl-, the molecule of the parent resorcarene 1 also adopts a boat conformation whose cavity is considerably extended by four hydrogen-bonded chloride anions. The pocket formed in this way again includes two 3+ ions as a result of electrostatic and hydrogen bonding host-guest interactions. All these structures show that the boat conformers of resorcarenes can be used as a novel motif for the construction of hydrogen-bonded assemblies capable of molecular inclusion and encapsulation. PMID- 11405474 TI - Ternary alkali metal transition metal acetylides A2MC2 (A = Na, K; M = Pd, Pt). AB - Ternary transition metal acetylides A2MC2 (A = Na, K; M = Pd, Pt) can be synthesised by reaction of the respective alkali metal acetylide A2C2 with palladium or platinum in an inert atmosphere at about 350 degrees C. The crystal structures are characterised by (infinity)1[M(C2)(2/2)2-] chains, which are separated by the alkali metals (P3m1, Z = 1). The refinement of neutron powder diffraction data gave C-C = 1.263(3) A for Na2PdC2 (Na2PtC2: 1.289(4) A), which is distinctively longer than the expected value for a C-C triple bond (1.20 A). On the basis of band-structure calculations this can be attributed to a strong back-bonding from the metal into the anti-bonding orbitals of the C2 unit. This was further confirmed by Raman spectroscopic investigations, which showed that the wavenumbers of the C-C stretching vibrations in Na2PdC2 and Na2PtC2 are about 100 cm(-1) smaller than in acetylene. 13C MAS-NMR spectra demonstrated that the acetylenic C2 units in the title compounds are very different from those in acetylene. Electrical conductivity measurements and band-structure calculations showed that the black title compounds are semiconductors with a small indirect band gap (approximately 0.2 eV). PMID- 11405475 TI - An unprecedented type of migratory insertion reactions of unsaturated C3 units into Rh-O and Rh-C bonds. AB - A series of iodo- and hydroxorhodium(I) complexes of the general composition trans-[RhX(=C=C=CRR')(PiPr3)2] (X = I: 5-7; X = OH: 8-11) was prepared from the related chlororhodium(I) precursors. The hydroxo compounds behave as organometallic Bronsted bases and react with acids like MeCO2H, PhCO2H, PhOH, or TsOH by elimination of water to give the substitution products trans [RhX'(=C=C=CRR')(PiPr3)2] (X' = MeCO2: 12, 13; X' = PhCO2: 14; X' = PhO: 15, 16; X' = TsO: 17, 18) in good to excellent yields. In contrast to the tosylates 17, 18, which react with CO by cleavage of the allenylidene-metal bond to give trans [Rh(OTs)(CO)(PiPr3)2] (19), treatment of the acetato and phenolato derivatives 12, 13 and 15, 16 with CO affords by migratory insertion of the allenylidene unit into the Rh-O bond the alkynyl complexes trans-[Rh[C(triple bond)CCR(R')X'](CO)(PiPr3)2] (X' = MeCO2: 20, 21; X' = OPh: 22, 23). Similarly, the reactions of the hydroxo compounds 8, 10, and 11 with CH2(CN)2 and either CO or CNMe yield the carbonyl and the isocyanide complexes trans-[Rh[C(triple bond)CCR(R')CH(CN)2](L')(PiPr3)2] (L' = CO: 25-27; L' = CNMe: 28-30), respectively. By protolytic cleavage of the Rh-C sigma bond the gamma functionalized alkynes HC(triple bond)CCR(R')CH(CN)2 (31, 32) are generated from 25, 26 and HCl in benzene. The molecular structure of 22 was determined by X-ray crystallography. PMID- 11405476 TI - Metal-modified nucleobase sextet: joining four linear metal fragments (trans a2PtII) and six model nucleobases to an exceedingly stable entity. AB - Crosslinking of three different model nucleobases (9-ethyladenine, 9-EtA; 9 ethylguanine, 9-EtGH; 1-methyluracil, 1-MeU) by two linear trans-aPtII (a = NH3 or CH3NH2) entities leads to a flat metal-modified base triplet, trans,trans [(NH3)2Pt(1-MeU-N3)(mu-9-EtA-N7,N1)Pt(CH3NH2)2(9-EtGH-N7)]3+ (4b). Upon hemideprotonation of the 9-ethylguanine base at the N1 position. 4b spontaneously dimerizes to the metalated nucleobase sextet 5, [(4b)(triple bond)(4b-H)]5+. In this dimeric structure a neutral and an anionic guanine ligand, which are complementary to each other, are joined through three H bonds and additionally by two H bonds between guanine and uracil nucleobases. Four additional interbase H bonds maintain the approximate coplanarity of all six bases. The two base triplets form an exceedingly stable entity (KD = 500 +/- 150 M(-1) in DMSO), which is unprecedented in nucleobase chemistry. The precursor of 4b and several related complexes are described and their structures and solution properties are reported. PMID- 11405477 TI - Experimental evidence supporting a CuIII intermediate in cross-coupling reactions of allylic esters with diallylcuprate species. AB - The reaction between an allylic ester and a magnesium diallylcuprate, or an allylic Grignard reagent in combination with a catalytic amount of a copper salt, has been studied. These reactions yield a mixture of homo- and cross-coupled 1,5 diene products. The product ratios obtained are close to those expected for a reaction proceeding via a triallylcopper(III) intermediate consisting of three equivalent allyl groups bound to copper. When the reaction is performed with a stoichiometric amount of a preformed diallylcuprate, a homo-coupling/cross coupling ratio larger than that predicted for a CuIII intermediate is observed. However, on dilution this ratio decreases and becomes close to the predicted ratio. The deviation from the predicted homo-coupling/cross-coupling ratio was accounted for by an olefin-induced homo-coupling, as demonstrated in control experiments. The possibility of the allylic ligands to coordinate to the metal center in a eta3 or eta1 fashion provides an opportunity for the stabilization of the intermediate CuIII species. PMID- 11405478 TI - Studies of the synthesis of SSZ-25 zeolite in a "mixed-template" system. AB - The synthesis of aluminosilicate zeolite, SSZ-25, is described using a two component organic guest molecule strategy. This method has been recently described by us and is quite effective in crystallizing a variety of aluminosilicate zeolites with reduction in template cost. In this instance the original organic guest component used in the discovery of SSZ-25, N',N',N' trimethyl-2-adamantammonium cation, is used in conjunction with a variety of smaller amines. The amine is the major organic component in the synthesis and the quaternary ammonium compound, while a minor component, is essential for structure direction to the desired zeolite product. Studies here show that the adamantyl component is preferentially taken up during crystal growth. Our studies showed that the use of the multi-organic component approach to synthesis resulted in a faster and cheaper route to crystallizing SSZ-25. The SSZ-25 has been described in the literature as having the MWW topology and as such has two different channel systems accessed by 10-ring openings; the channel systems do not intersect. One set of 10-rings open into large cavities. In this study we used NMR spectroscopy to examine the as-made materials and to see if the two organic components are used in different capacities in the synthesis, probing if the admantyl component is only found in these larger cavities. The crystalline products could be affected by solvent extraction with polar solvents like dimethylformamide: the X-ray powder patterns and elemental analyses changed with solvent treatment. The solvent-treated crystals were studied by NMR spectroscopy, elemental analysis, and argon adsorption. The two-component organic guest approach was also found to be quite flexible not only with regard to the amine, but also towards the quaternary ammonium compounds. Non-adamantyl polycyclic templates could be used, and polar but non-quaternized adamantyl derivatives also succeeded in directing towards SSZ-25 formation. On the other hand. in one instance it was shown that the two-component approach favors SSZ-25 as the kinetic product but at longer run times Ostwald ripening was observed, leading to the disappearance of SSZ-25 and formation of ferrierite zeolite and quartz. PMID- 11405479 TI - Synthesis and structural characterization of the novel cluster compound. AB - Reaction of [Mo3Y(mu-S)3(dtp)4(H2O)] (Y = O, S; dtp = S2P(OC2H5)2(-)) with HgI2 gave the novel compound [[Mo3S7(dtp)3]4 x I][(HgI3)3] x 4H2O (1), which contains a [[Mo3S7(dtp)3]4 x I] tetramer and (HgI3)-. Compound 1 has been characterized by IR, Raman, UV/Vis, and NMR spectroscopy and single-crystal X-ray diffraction analysis. It is shown that this formation process can be referred to as a new cluster reaction. The structure and spectroscopic data of the tetramer is also compared with that of the related discrete cluster [Mo3S7(dtp)3 x I]. Crystal data: space group F23, a = 26.786(3) A, V = 19218.7(4) A3, Z = 4, R = 0.059. PMID- 11405480 TI - Nitronyl and imino nitroxides: improvement of Ullman's procedure and report on a new efficient synthetic route. AB - The synthesis of nitronyl and imino nitroxides has been reexamined with the aim of both increasing yields and of offering opportunities for new structures. The conditions for the formation of 2,3-bis(hydroxyamino)-2,3-dimethylbutane, the key intermediate of Ullman's route, have been carefully studied, and a new procedure is proposed, which affords the free base in a very pure form and up to 60% yield. Full characterization of this intermediate including an X-ray crystal structure is presented. An alternative synthetic route through 2,3-diamino-2,3 dimethylbutane and the corresponding imidazolidines which bypasses the delicate synthesis of the bis(hydroxyamino) compound is described. It is shown that 3 chloroperbenzoic acid is an effective oxidant for the transformation of adequately substituted imidazolidines into nitronyl nitroxides, which are obtained in high yield. An illustration of the potentialities of this new route, a new nitronyl nitroxide with two ethyl substituents in positions 4 and 5 of the imidazoline ring, is reported. The scope and limitations of the two routes are discussed. PMID- 11405481 TI - A new family of chelating diphosphines with a transition metal stereocenter in the backbone: novel applications of "chiral-at-rhenium" complexes in rhodium catalyzed enantioselective alkene hydrogenations. AB - The title compounds are accessed by sequences starting with racemic and enantiomerically pure [(eta5-C5H5)Re(NO)(PPh3)(CH3)]. Reactions with chlorobenzene/HBF4, PPh2H, and tBuOK give the phosphido complex [(eta5 C5H5)Re(NO)(PPh3)(PPh2)] (3). Reactions with Ph3C+ BF4-, PPh2H, and tBuOK give the methylene homologue [(eta5-C5H5)Re(NO)(PPh3)(CH2PPh2)] (9). Treatment of 3 or 9 with nBuLi or tBuLi and then PPh3Cl gives the diphosphido systems [(eta5 C5H4PPh2)Re(NO)(PPh3)((CH2)nPPh2)] (n = 0/1, 5/11). Reactions of 5 and 11 with [Rh(NBD)Cl]2/AgPF6 (NBD = norbornadiene) give the rhenium/rhodium chelate complexes [(eta5-C5H4PPh2)Re(NO)(PPh3)((mu-CH2)nPPh2)Rh(NBD)]+ PF6- (n = 0/1, 6+/12+ PF6-; 30-32% overall from commercial Re2(CO)10). The crystal structures of 6+ PF6- and 12+ PF6- are compared to those of 3 and 9, and other rhodium complexes of chelating bis(diphenylphosphines). The chiral pockets defined by the PPh2 groups show unusual features. Four alkenes of the type (Z) RCH=C(NHCOCH3)CO2R' are treated with H2 (1 atm) and (R)-6+ PF6- or (S)-12+ PF6- (0.5 mol%) in THF at room temperature. Protected amino acids are obtained in 70 98% yields and 93-82% ee [(R)-6- PF6-] or 72-60% ee [(S)-12+ PF6-]. Pressure and temperature effects are defined, and turnover numbers of > 1600 are realized. PMID- 11405482 TI - Phosphinofenchol or metastable phosphorane? Phosphorus derivatives of fenchol. AB - Not the expected phosphinofenchol 1 but phosphorane 2 is obtained after reaction of 2-lithio(diphenylphosphino)benzene with (-)-fenchone. Surprisingly, ONIOM(B3LYP/6-31G*:UFF) computations of 1 and 2 as well as B3LYP analyses of smaller model systems point to a lower thermodynamic stability of phosphoranes relative to their isomeric alkoxyphosphines. An analogue inherent instability is computed for the methylphosphorane 10, which is also synthesized and characterized by X-ray analysis. Decreasing ring size in cyclic phosphoranes, that is, from five- to four-membered ring systems, destabilizes cyclic phosphoranes even more. This computational prediction is verified experimentally by reaction of lithiomethyl(diphenylphosphine) with (-)-fenchone and subsequent isolation of the corresponding phosphinofenchol. Protonation or alkylation of phosphoranide intermediates can account for the formation of metastable phosphoranes. PMID- 11405483 TI - Diastereoselective formation of host-guest complexes between a series of phosphate-bridged cavitands and alkyl- and arylammonium ions studied by liquid secondary-ion mass spectrometry. AB - The reactions occurring between a new class of cavitands that carry up to four dioxaphosphocin binding units and alkyl- and arylammonium ions was investigated by liquid secondary-ion mass spectrometry (LSIMS). As the cavitands existed as distinct diastereomers with different spatial orientation of their binding groups, these geometrical differences proved to have a dramatic influence on their chemical properties, including their ability to form host-guest complexes. In practice, only the cavitands that carry at least three P=O groups oriented toward the inside of the cavity were demonstrated to be strong ligands toward organic ammonium ions, whereas those with only two converging binding groups (either adjacent or opposite in the cavitand structure) still formed host-guest complexes, but they were much weaker. Adjacent binding sites proved to be more effective in interacting with organic ammonium ions than those lying in opposite positions. The isomers with no converging P=O groups did not act as molecular receptors. Even the isomer with one group oriented toward the inside of the cavity did not form host-guest complexes, as the absence of synergistic hydrogen bonding made the interaction from inside the cavity energetically equivalent (or even less favorable) to the outside binding. The presence in the cavitand structure of substituents with an electron-donating character proved to increase the proton affinity of the P=O binding groups and, consequently, their binding energy. The strong proton affinity of the cavitands led to the formation of stable host-guest complexes, as confirmed by the collisionally activated dissociation experiments. Effects of steric hindrance were weak, at least for the cavitands with three converging P=O groups. This confirmed that the cavity has a wide and readily accessible opening. The relative complexation constants were measured for the various guests, yielding scales of relative affinity toward each cavitand. These relative constants may represent thermodynamic values referred to the matrix used in LSIMS experiments, namely 3-nitrobenzyl alcohol (NBA), provided that kinetically controlled selvedge processes are negligible. Absolute complexation constants could not be obtained on account of the unknown pH and the protonation constant in the NBA matrix. PMID- 11405484 TI - Long-term retention of a novel antioxidant sulphur-substituted fatty acid analogue after local delivery in porcine coronary arteries. AB - OBJECTIVE: Antioxidants have been suggested to reduce restenosis after balloon angioplasty. A novel sulphur-containing fatty acid, tetradecylthioacetic acid (TTA), with antioxidant properties, is efficiently incorporated into cellular phospholipids. We have determined the uptake and retention of TTA after local coronary artery delivery in 20 pigs. DESIGN: Radiolabelled TTA was delivered to 40 main coronary arteries via a multiporous coronary angioplasty balloon catheter inflated before, after, or without overstretch vessel injury. The animals were killed at intervals of up to 6 weeks post-procedure. The radioactivity of the tissue sections was determined as nmol TTA/g tissue. RESULTS: Concentrations of TTA in the coronary arteries were 1.84 +/- 0.45 nmol/g up to 24 h, 1.50 +/- 0.96 nmol/g at 2 weeks, 0.22 +/- 0.11 nmol/g at 4 weeks and a trace was present at 6 weeks (p-value for trend <0.01). The arterial wall uptake at the delivery site was higher than distal to delivery (1.84 +/- 0.37 vs 0.55 +/- 0.13 nmol/g, p = 0.006) and perivascular fat (p < 0.01) but not higher than in the myocardium. Infusion before, after or without vessel injury was not important for tissue concentration. CONCLUSIONS: After local coronary artery delivery, the antioxidant TTA is taken up by the arterial wall in which it is retained for at least 4 weeks. PMID- 11405485 TI - Self-management of oral anticoagulant therapy for mechanical heart valve patients. AB - OBJECTIVE: Self-management of oral anticoagulant therapy (OAT) has shown good results on a short-term basis. We hypothesize that self-management of OAT provides a better quality of treatment than conventional management also on a long-term basis. The aim of this study was to assess the quality of self management of OAT in patients with mechanical heart valve prostheses on a 4-year perspective in a prospective, non-randomized study. DESIGN: Twenty-four patients with mechanical heart valves and on self-managed OAT were followed for up to 4 years. A matched, retrospectively selected group of conventionally managed heart valve patients (control group) was used as reference. RESULTS: The median observation time was 1175 days (range: 174-1428 days). The self-managed patients were within therapeutic INR target range for a mean of 78.0% (range: 36.1%-93.9%) of the time compared with 61.0% (range 37.4%-2.9%) for the control group. CONCLUSION: Self-management of OAT is a feasible and safe concept for selected patients with mechanical heart valve prostheses also on a long-term basis. It provides at least as good and most likely better quality of anticoagulant therapy than conventional management assessed by time within the therapeutic International Normalized Ratio (INR) target range. PMID- 11405486 TI - Aortic valve replacement with pericardial valves in patients with small aortic roots. Clinical results in a consecutive series of patients receiving 19 and 21 mm prostheses. AB - OBJECTIVE: To determine how second generation pericardial valves perform in patients with small aortic roots. DESIGN: Ninety patients who underwent isolated aortic valve replacement (AVR) with 19 or 21 mm Mitroflow or Carpentier-Edwards (Perimount) valves between 1989 and 1996 were studied. Mean age was 78 years. Concomitant coronary bypass surgery was performed in 41%. RESULTS: Thirty-day mortality was 5.6%. Ninety-seven percent had acceptable transprosthetic mean pressure gradients (25 mmHg or less) 1 week after surgery. Follow-up was 100% complete and 76% of the patients were alive after a mean of 5 years. There was no structural valve failure or valve thrombosis. One patient required reoperation for perivalvular leak. Four patients had transient ischemic attacks and seven had strokes. These figures are, however, within the expected range for the age. CONCLUSION: Second generation pericardial valves perform well in elderly patients with small aortic roots. Postoperative hemodynamics are acceptable, valve durability of up to 8 years adequate, and the clinical results good, considering the age of the patients. PMID- 11405487 TI - Left atrial appendage outflow velocity index is superior to conventional criteria for prediction of maintenance of sinus rhythm after cardioversion. An echocardiographic study in patients with atrial fibrillation of a few months' duration. AB - OBJECTIVE: To investigate whether left atrial appendage outflow velocity alone or in relation to left atrial diameter is a superior predictor of sinus rhythm maintenance after cardioversion compared with traditional clinical or echocardiography parameters. DESIGN: Sixty-two patients with their first episode of atrial fibrillation were examined using echocardiography before DC cardioversion. At one month's follow-up, 42 patients had maintained sinus rhythm (group A), and 20 had relapsed into atrial fibrillation (group B). There were no differences in arrhythmia duration or antiarrhythmic therapy between the groups. RESULTS: Left atrial diameter measured by echocardiography was smaller in group A (42 mm, 95% CI 40.9-44.1 mm) compared with group B (46 mm, 95% CI 43.4-48.2, p < 0.05). Patients in group A had a higher left atrial appendage outflow velocity at 0.44 m/s (95% CI 0.39-0.49) compared with 0.34 m/s (95% CI 0.30-0.37) in group B (p < 0.01). The ratio of left atrial appendage flow to left atrial diameter was 0.011 (95% CI 0.009-0.012) in group A compared with 0.008 (95% CI 0.007-0.009) in group B, and 63% (95% CI 33-78) of the patients in group A had velocity ratio >0.009 compared with 20% (95% CI 2-38) in group B, (p < 0.01). Stepwise multiple logistic regression analysis showed that a velocity ratio >0.009 was the only predictor for maintenance of sinus rhythm one month after cardioversion with an odds ratio of 6.4 (95% CI 1.9-23.8), (p = 0.004). CONCLUSION: The ratio of left atrial appendage outflow velocity to left atrial diameter is superior to the traditionally used criteria for prediction of maintenance of sinus rhythm following DC-conversion of first-episode atrial fibrillation. PMID- 11405488 TI - Vectorcardiography is superior to conventional ECG for detection of myocardial injury after coronary surgery. AB - OBJECTIVE: The reliability of conventional scalar ECG for diagnosis of perioperative myocardial infarction (PMI) in cardiac surgery has been questioned. For the diagnosis of myocardial infarction in general vectorcardiography (VCG) is superior to ECG. Therefore, the usefulness of conventional VCG and computerized analysis of spatial VCG changes for diagnosis of PMI were studied. DESIGN: VCG registrations were obtained from 218 patients undergoing coronary surgery. The spatial QRS vector loop area of each VCG registration was calculated and the loop area before surgery compared with the loop area after surgery. Conventional VCG criteria for myocardial infarction and set values for loop area reduction were related to sustained elevation of plasma troponin-T and clinical course. RESULTS: Both conventional VCG criteria and spatial changes translated better than Q-waves on scalar ECG into elevation of biochemical markers of myocardial injury and impaired clinical course. CONCLUSION: VCG appears superior to conventional ECG as regards detection of myocardial injury in coronary surgery. Computerized programs have facilitated the registration and the interpretation of VCG and this methodology deserves further evaluation in cardiac surgery. PMID- 11405489 TI - Correlation between a mid-ventricular volume segment and global left ventricular volume measured by the conductance catheter. AB - OBJECTIVES: To investigate whether acute volume changes in single volume segments of the left ventricle can be correlated with global volume changes. If so, changes in global volume might be predicted from changes in segmental volumes. DESIGN: Volume changes were recorded in six pigs in five intraventricular segments, from apex to heart base, using the conductance catheter (at baseline, after 60 min of apical ischaemia, during preload reduction and afterload increase). A computer algorithm was created to calculate the instantaneous absolute difference between the curve shape of global and normalized segmental volume as a percentage of global stroke volume. RESULTS: For a mid-cardiac volume segment constituting 34 (14-39)% [median (range)] of global stroke volume, the mean difference over a cardiac cycle was 4 (1-8)% at baseline. Apical ischaemia resulted in apical dyskinesia, but did not influence the mid-cardiac segment. CONCLUSIONS: The volume curve from a segment at mid-cardiac level seems to be a good estimator of the global volume curve, thus giving a foundation for estimation of global volume changes from such a segment. PMID- 11405490 TI - Right heart assist ensures hemodynamic stability during beating heart coronary surgery on marginal arteries. An animal experimental porcine study. AB - OBJECTIVES: In many heart centers myocardial revascularization using beating heart coronary surgery has partly replaced conventional coronary artery bypass grafting (cCABG) using cardiopulmonary bypass. However, access to the marginal arteries is problematic and hampered by space limitations, which might compromise the quality of the anastomoses and it entails a significant risk of hemodynamic instability subsequent to the manipulations of the heart. Hemodynamic impairment may be caused by dislocation of the low-pressure right atrium and ventricle. Therefore, it was hypothesized that the use of Right Heart Assist (RHA) may be able to ensure hemodynamic stability when tilting and manipulating the heart. DESIGN: In an animal experimental model RHA was implemented in order to study the hemodynamic impact of dislocating the heart in a standardized fashion. RESULTS: Dislocation of the unassisted heart to expose the first and second marginal branches caused a decline in arterial blood pressure, cardiac output and venous saturation of 38-48%. Supported by RHA this decrease was minimized to 0-17%. CONCLUSION: RHA for beating heart procedures indicates several advantages: ample time and safety while making the anastomoses at the marginal branches, less space limitations and enhanced control of hemodynamic stability compared with no RHA. This operation technique is challenging current practice and indications, and may potentially replace cCABG for the majority of patients. PMID- 11405491 TI - Cytokine responses in patients undergoing coronary artery bypass surgery after ischemic preconditioning. AB - OBJECTIVE: The release of proinflammatory cytokines has been shown to be associated with the development of complications after coronary artery bypass grafting with cardiopulmonary bypass. The purpose of the present study was to establish whether ischemic preconditioning (IP) could limit inflammatory cytokines release in patients undergoing elective coronary artery bypass surgery. METHODS: Twenty-two patients with multiple-vessel coronary artery disease and stable angina admitted for first-time elective coronary artery bypass surgery were randomized into control or ischemic preconditioning groups. Patients in the IP group were exposed to two cycles of two-minute myocardial ischemia, followed by three minutes of reperfusion, at the beginning of the revascularization operation, before the cross-clamping and ischemic period used for coronary artery bypass graft anastomosis. Peripheral plasma levels of TNF-alpha, IL-6, IL-8 and IL-10 were measured perioperatively. RESULTS: Significant elevation of IL-6, IL-8 and IL-10 were observed in both groups after reperfusion. Ischemic preconditioning has no effect on cytokine release in the early stage after reperfusion. Arterial blood IL-6 levels in the preconditioning group were significantly lower than in controls at 20 h after declamping (52.93 +/- 9.79 vs 96.04 +/- 17.56 pg/ml, p < 0.05). CONCLUSIONS: The results indicate that ischemic preconditioning results in no effect on systemic inflammatory cytokine release in the early stage but a delayed reduction in IL-6 levels at 20 h after reperfusion. PMID- 11405492 TI - Increased FiO2 improves intrathecal oxygenation during thoracic aortic cross clamping in pigs. AB - OBJECTIVES: To investigate the effect of 100% oxygen ventilation on cerebrospinal fluid (CSF) oxygenation in 11 pigs during thoracic aortic cross-clamping. DESIGN: An aorto-aortic shunt was used for control of central hemodynamics and study of hypoperfusion by exsanguination. CSF PO2, PCO2 and pH were continuously monitored before and during clamping. The changes in hemodynamic parameters and intrathecal gas tensions in response to variations in proximal mean aortic pressure and fraction of inspired oxygen (FiO2) were recorded. RESULTS: Baseline CSF PO2 decreased from 4.8 +/- 1.9 to 2.6 +/- 2.2 kPa following aortic occlusion. Increasing FiO2 to 1.0 resulted in a significant increase in CSF PO2 to 4.1 +/- 3.0 with a return to 2.7 +/- 2.1 kPa after reducing FiO2 to 0.4 again. The same variations in FiO2 did not induce any significant changes in CSF PO2 during hypotension. CONCLUSION: Increased FiO2 during experimental thoracic aortic cross clamping with stable proximal arterial pressure helps to maintain CSF PO2, whereas severe hypotension could not be compensated for by hyperoxemia. PMID- 11405493 TI - Bilateral chylothorax caused by pleuropulmonary lymphangiomyomatosis: a challenging problem in thoracic surgery. AB - Massive left-sided pleural effusion in a 35-year-old man was initially diagnosed as idiopathic spontaneous chylothorax and treated with serial thoracenteses and left thoracotomy. Six weeks later, a right thoracotomy was performed for contralateral chylothorax, and histologic examination revealed lymphangiomyomatosis. The patient survived this rare and potentially fatal disease. We have found no previously published case of bilateral lymphangiomyomatosis treated with separate thoracotomies because of bilateral chylothorax. PMID- 11405494 TI - Subacute left ventricular free wall rupture. AB - In a 59-year-old man, Left ventricular free wall rupture following acute myocardial infarction was diagnosed by transthoracic echocardiography, left ventriculography and a combination of saline injection into the left ventricle and concomitant transthoracic echocardiography. The Operation was successfully performed with an extracorporeal bypass on the beating heart. Some technical aspects of the treatment are discussed. PMID- 11405495 TI - Extralobar pulmonary sequestration in the posterior mediastinum. AB - Extralobar pulmonary sequestration was incidentally detected in a 65-year-old man. Preoperatively, the lesion presented as a posterior mediastinal mass. At operation the mass was found to be attached to the paravertebral region at the level of the seventh thoracic vertebra by a feeding artery originating from the descending aorta. Only two cases of extralobar pulmonary sequestration in the posterior mediastinum have been previously reported. PMID- 11405496 TI - Surgical treatment of aortopulmonary window with tetralogy of Fallot. AB - An association between aortopulmonary window and tetralogy of Fallot is rare. We report a case of the combined anomalies in which, possibly for the first time, the diagnosis was made by echocardiography alone, and in which successful surgical treatment was performed in early infancy. PMID- 11405497 TI - Intravascular ultrasound improves the quality of coronary interventions, but is it worth the extra time and money invested? PMID- 11405498 TI - Mechanical valve anticoagulation--between Scylla and Charybdis. PMID- 11405499 TI - Vectorelectrocardiography in coronary artery disease. AB - Continuous ST-segment monitoring by either continuous 12-lead ECG or continuous vectorcardiography provides reliable information regarding ST changes in patients with ongoing myocardial ischemia with or without concurrent chest pain. ST segment monitoring enables the clinician to continuously follow the dynamic changes that characterize unstable angina and acute myocardial infarction syndromes. It provides important information for risk stratification in unstable coronary syndromes and helps in differentiating between extra-cardiac chest pain and acute coronary disease. The use of VECG for detection of perioperative myocardial damage is promising but much work still needs to be done to clarify the prognostic and clinical value of VECG in this setting. PMID- 11405500 TI - Intensive use of intravascular ultrasound during coronary angioplasty. A six month campaign. AB - BACKGROUND: Compared to coronary angiography, intravascular ultrasound (IVUS) gives additional information important for the percutaneous transluminal coronary angioplasty (PTCA) procedure, but is time-consuming and may cause complications. AIM: To evaluate, during a period of intensive use of IVUS, the impact of IVUS on the final decision on balloon/stent diameter, consumption of devices, time consumption and IVUS-related complications. METHOD: During a 6-month period, IVUS was contemplated in all PTCA procedures and the reason for not using IVUS was specified. We used CVIS during the first, and Endosonics during the last 3 months, and both periods started with 1 week of hands-on practice. All procedures were to be planned according to an initial quantitative coronary angiography (QCA), and the finally achieved result, material used and complications were registered. RESULTS: The proportion of IVUS/PTCA was 37% during, 8% 6 months before and 12% 6 months after the study period. Three hundred and twenty-three patients were included in the study (57% of all patients), 199 of them were subjected to IVUS. The indications for PTCA during the study period were stable angina (58%), unstable angina (32%) and acute myocardial infarction (10%). The main reasons for not doing IVUS were use of 6F guiding catheter (13%), urgent procedure (12%) and occluded vessel (11%). Initial QCA detected 253 stenoses in 199 patients and 64 additional stenoses were treated, most of them probably detected by IVUS. QCA systematically underestimated vessel size, particularly in small vessels. There was a non-significant trend to more accurate estimations towards the end of the study in small vessels. Dissection, probably due to IVUS, occurred in two cases (1%). There were no significant differences in the number of devices used in IVUS compared to non-IVUS patients. The procedural time was 24 min longer in IVUS than in non-IVUS cases and more stenoses were treated per procedure in the IVUS group. CONCLUSION: Coronary angiography often underestimated balloon/stent size but in an unpredictable way, with a substantial proportion of significant stenoses being undetected. IVUS had few serious complications, did not increase device consumption but prolonged procedural time. PMID- 11405501 TI - Cost-effectiveness analysis of intravascular ultrasound guided percutaneous coronary intervention versus conventional percutaneous coronary intervention. AB - OBJECTIVE: Investigation of the cost-effectiveness of intravascular ultrasound (IVUS) guided percutaneous coronary intervention (PCI) compared to PCI guided by coronary angiography (CAG). METHODS: One hundred and eight men referred for PCI, were randomized to IVUS or CAG guided PCI. After 6 months, the patients were subjected to a study related clinical and invasive follow-up investigation by CAG, IVUS and intracoronary Doppler flow measurements. Incremental costs of IVUS guided procedures and costs of re-interventions were estimated using the Activity Based Costing (ABC) method. RESULTS: Patients randomized to IVUS guided PCI experienced an improved clinical outcome, with lower angina levels than patients in the CAG guided group. The initial cost of performing IVUS guidance was increased due to extra procedure time, IVUS catheters and slightly more balloons and stents, but fewer patients in the IVUS guided group needed re-intervention. Overall, these savings outweighed the initial cost increase. CONCLUSION: Our data suggest that when performing IVUS guided PCI, costs as well as benefits increase. The increased benefits measured as cost savings resulting from less restenosis outweigh the cost increase from performing the IVUS guided PCI as opposed to CAG guided PCI. PMID- 11405502 TI - Predictors of restenosis after coronary angioplasty. A study on demographic and metabolic variables. AB - OBJECTIVE: The major concern about percutaneous transluminal coronary angioplasty (PTCA) is the high incidence of restenosis. METHODS: Demographic, clinical and biochemical data were recorded 2 weeks prior to PTCA in 388 patients fulfilling the criteria for initial stenosis, successful PTCA, and angiographic follow-up after 6 months. Restenosis was evaluated by quantitative coronary angiography. RESULTS: Variables predictive of restenosis in univariate analysis were diabetes mellitus, male gender, and the levels of high density lipoprotein (HDL) cholesterol, apolipoprotein A1 (Apo A1) and thio-barbituric acid-reactive substances (TBARS). In trend analysis through quartiles TBARS and fasting glucose levels were significantly associated with restenosis (p = 0.016 and 0.044, respectively), whereas the negative predictivity of Apo A1 and HDL-cholesterol were of borderline significance. In multivariate analysis male gender and diabetes mellitus showed predictivity of significance, and a negative predictivity was also apparent for HDL-cholesterol. CONCLUSION: We conclude that diabetes mellitus, male gender, and low HDL-cholesterol are predictors of restenosis 6 months after PTCA. In addition, TBARS may be a marker for the development of restenosis after PTCA. PMID- 11405503 TI - Thrombolytic therapy preserves vagal activity early after acute myocardial infarction. AB - OBJECTIVE: The purpose of this study was to evaluate the effects of thrombolytic therapy on vagal tone after acute myocardial infarction (AMI). DESIGN: Holter monitoring for 24 h was performed at hospital discharge and 6 weeks after AMI in 74 consecutive male survivors of a first AMI, who fulfilled established criteria for thrombolytic therapy. Thirty-five patients received thrombolyses, while the remaining 39 patients did not (controls). In each Holter recording 24-h heart rate variability was calculated as pNN50, which represents the percentage of successive RR interval differences >50 ms. Alterations in pNN50 are known to reflect changes in vagal tone. RESULTS: The analysis showed that controls early after AMI had low pNN50 values without any diurnal changes. Six weeks after AMI pNN50 values in controls exhibited a circadian rhythm with higher values during night-time. This pattern was similar to the pattern observed in thrombolysed patients early after AMI. In thrombolysed patients pNN50 values, particularly at night, were further improved 6 weeks after AMI (p = 0.037). CONCLUSION: These observations indicate that thrombolytic therapy, given for a first AMI, preserves vagal activity when compared with patients who are not thrombolysed. The enhanced parasympathetic tone may be a part of the beneficial mechanisms responsible for the reduction in mortality after thrombolysis in AMI. PMID- 11405504 TI - Cholesterol efflux capacity in vitro predicts the severity and extent of coronary artery disease in patients with and without type 2 diabetes. AB - OBJECTIVE: To investigate the relation between severity and extent of coronary artery disease (CAD) and in vitro cholesterol efflux capacity. DESIGN: This study consisted of 46 type 2 diabetic, and 42 nondiabetic men undergoing coronary angiography. Quantitative coronary angiography was used to estimate the severity, extent, and overall "atheroma burden" of CAD. The capacity of patient plasma to induce cholesterol efflux from cultured Fu5AH rat hepatoma cells was measured in vitro. RESULTS: In the combined study population (n = 88), there was a significant inverse correlation between efflux and global atheroma burden (r = 0.23, p < 0.05). In the diabetic group, the global atheroma burden index was independently associated both with cholesterol efflux and with LpA-I levels. However, in the nondiabetic CAD group this association was lost when efflux and LpA-I levels were included in the same model. CONCLUSION: The present study demonstrated that efflux capacity was inversely associated with the severity and extent of CAD. In the diabetic group this association was independent of LpA-I levels, suggesting impaired antiatherogenic potential of these particles in type 2 diabetic patients. PMID- 11405505 TI - The new European Respiratory Journal: alive and kicking. PMID- 11405506 TI - Why a series on imaging in the ERJ? PMID- 11405507 TI - The Bronchitis Randomized On NAC Cost-Utility Study (BRONCUS): hypothesis and design. BRONCUS-trial Committee. AB - Chronic obstructive pulmonary disease (COPD) is an irreversible disorder characterized by airflow obstruction and a progressive decline in forced expiratory volume in one second (FEV1). At present, no treatment except quitting smoking appears to affect the progression of the disease. Oxidative stress has been implicated in its pathogenesis. The Bronchitis Randomized on NAC Cost Utility Study (BRONCUS) is a phase III, randomized, double-blind, placebo controlled, parallel group, multicentre study designed to assess the effectiveness of the antioxidant agent N-acetylcysteine (NAC) in altering the decline in FEV1, exacerbation rate, and quality of life in patients with moderate to severe COPD. In addition, cost-utility of the treatment will be estimated. Patients will be followed for 3 yrs and evaluated every 3 months. The necessary sample size to demonstrate an effect on the decline in FEV1 of 20 mL x yr(-1) was estimated to be 478 patients. Five hundred and twenty-three patients with moderate to severe COPD were recruited from 10 European countries from June 1, 1997-December 31, 1999. They were 63+/-8 yrs old and consisted of 243 (46%) current smokers and 280 (54%) exsmokers. Patients had on the average 4.9+/-1.6 exacerbations during the last 2 yrs. Postbronchodilator FEVI averaged 57+/-9% and the reversibility after 400 microg of Salbutamol averaged 4+/-4% predicted. The final results of the trial will be available in about 2 yrs. The study will provide objective data on the effects of N-acetylcysteine on outcome variables in chronic obstructive pulmonary disease. PMID- 11405508 TI - Dose dependent increased mortality risk in COPD patients treated with oral glucocorticoids. AB - Systemic corticosteroids are often administered in COPD patients. The relationship between systemic glucocorticoids and mortality in patients with moderate to severe chronic obstructive pulmonary disease (COPD) was retrospectively analysed. Baseline characteristics of the patients, in stable clinical condition, were collected on admission to a pulmonary rehabilitation centre. Overall mortality was asessed at the end of follow-up. The Cox proportional hazards model was used to quantify the relationship between glucocorticoid use, distinguishing administration route (oral/inhalation) and oral dose, and overall mortality, adjusted for the influence of age, sex, smoking, lung function, resting arterial blood gases and body mass index. On multivariate analysis, oral glucocorticoid use at a (prednisone equivalent) dose of 10 mg x day(-1) without inhaled glucocorticoids, was associated with an increased risk (RR=2.34, 95% confidence interval (CI) 1.24-4.44) while 15 mg x day(-1) carried a relative risk of 4.03, CI = 1.99-8.15). A significant interaction was observed between inhaled and oral glucocorticoid use. Combined with inhaled glucocorticoids, the relative risk of oral glucocorticoid use appeared to be significantly smaller. It is concluded that in severe chronic obstructive pulmonary disease, maintenance treatment with oral glucocorticoids is associated with increased mortality in a dose-dependent manner. Since the present study design cannot exclude the possibility of bias by indication, further prospective studies are indicated using a broader patient characterization. PMID- 11405509 TI - Audit of acute admissions of COPD: standards of care and management in the hospital setting. AB - Despite publication of several management guidelines for COPD, relatively little is known about standards of care in clinical practice. Data were collected on the management of 1400 cases of acute admission with Chronic Obstructive Pulmonary Disease in 38 UK hospitals to compare clinical practice against the recommended British Thoracic Society standards. Variation in the process of care between the different centres was analysed and a comparison of the management by respiratory specialists and nonrespiratory specialists made. There were large variations between centres for many of the variables studied. A forced expiratory volume in one second measurement was found in only 53% of cases. Of the investigations recommended in the acute management arterial blood gases were performed in 79% (interhospital range 40-100%) of admissions and oxygen was formally prescribed in only 64% (range 9-94%). Of those cases with acidosis and hypercapnia 35% had no further blood gas analysis and only 13% received ventilatory support. Long-term management was also deficient with 246 cases known to be severely hypoxic on admission yet two-thirds had no confirmation that oxygen levels had returned to levels above the requirements for long-term oxygen therapy. Only 30% of current smokers had cessation advice documented. To conclude, the median standards of care observed fell below those recommended by the guidelines. The lowest levels of performance were for patients not under the respiratory specialists, but specialists also have room for improvement. The substantial variation in the process of care between hospitals is strong evidence that it is possible for other centres with poorer performance to improve their levels of care. PMID- 11405510 TI - Utility of echocardiography in assessment of pulmonary hypertension secondary to COPD. AB - Studies have confirmed the close correlation of echocardiographically estimated pulmonary arterial pressure with invasive measurements in patients with chronic obstructive pulmonary disease (COPD), but there are few data on utility of echocardiographic measurement in assessing pulmonary arterial hypertension (PH) in COPD and correlation with pulmonary function tests. Presence or absence of tricuspid regurgitation (TR) was determined by Doppler echocardiography in 73 consecutive COPD patients attending a hospital outpatient clinic. Transtricuspid pressure gradient (TTPG) was calculated. PH was defined as TTPG > or =30 mmHg. Patients also underwent spirometry, forced expiratory volume in one second (FEV1), single breath gas transfer (carbon monoxide transfer coefficient; (K(CO)) and carbon monoxide diffusing capacity of the lung; D(L,CO)) and arterial blood gas measurement. Measurable TR was observed in 56/73 patients (77%). There were no differences between the group in which TR was observed compared to that in which it was absent, with regard to age, smoking history nor pulmonary function variables. PH was seen in 31/56 cases (55%), with good reproducibility. There were statistically significant correlations of TTPG with FEV1 (r=-0.26, p=0.05), Kco (r=-0.31, p=0.04) and D(L,CO) (r = -0.42, p = 0.006) expressed as % pred. Stepwise regression analysis showed that age and K(CO) combined provide a multivariate model for prediction of TTPG. It is concluded the presence and degree of pulmonary arterial hypertension is readily and reliably determined by echocardiography in the majority of chronic obstructive pulmonary disease patients. Pulmonary arterial hypertension is common in severe chronic obstructive pulmonary disease and transtricuspid pressure gradient correlates with spirometry and indices of gas transfer, similar to previous invasive studies. In view of the adverse effects of pulmonary arterial hypertension on morbiditv and mortality routine echocardiography in patients with severe chronic obstructive pulmonary disease may be warranted. PMID- 11405511 TI - Alpha-1-antitrypsin genotyping with mouthwash specimens. AB - Alpha1-antitrypsin (alpha1-AT) deficiency is diagnosed as a two-stage procedure (concentration and phenotype). However the latter does not provide clues to the presence of null genes without family studies and obtaining blood from patients at a distance often proves difficult. The aim of the study was to assess the feasibility of genotyping alpha1-AT using buccal cells. Mouthwash specimens were sent by 84 patients (with a variety of phenotypes of alpha1-antitrypsin) through the post. Deoxyribonucleic acid (DNA) was isolated from buccal cells in each sample and subjected to polymerase chain reaction (PCR) using a genotyping kit to detect the S and Z alleles. Eighty-three of 84 samples received were suitable for amplification. The specific primers successfully identified the S and Z alleles in each case. However, five of the 35 samples obtained from patients thought to be Z allele homozygotes were found to be heterozygotes for another severe deficiency allele. These data confirm the feasibility of "at distance" testing for alpha1-antitrypsin deficiency alleles using buccal cells from mouthwash samples. The results raise the possibility that other deficiency alleles are more common than has previously been suspected. PMID- 11405512 TI - Erythromycin attenuates an experimental model of chronic bronchiolitis via augmenting monocyte chemoattractant protein-1. AB - The mechanisms underlying the therapeutic efficacy of erythromycin (EM) in diffuse panbronchiolitis (DPB) was investigated. For this purpose, an experimental rabbit model of DPB induced by Pseudomonas aeruginosa inoculation was employed. Daily administration of EM (3 mg x kg x day(-1)) led to an increase in the number of macrophages in bronchoalveolar lavage fluid (BALF) at an early phase, while reducing the size of granulomatous lesions at the late phase without affecting the number of viable bacteria recovered from the infected lung. Reverse transcriptase polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) and immunohistochemical studies showed that monocyte chemoattractant protein (MCP)-1 was produced in both BALF and infected lung. EM treatment resulted in a significant increase in the level of MCP-1 in BALF, while reducing that of tumour necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-8. EM also increased MCP-1 messenger ribonucleic acid (mRNA) and protein expression in the infected lung. MCP-1 blockade abolished the protective effect of EM, as neutralization of MCP-1 with anti-MCP-1 antibodies reduced the EM induced increase in the number of macrophages in BALF, and augmented size of the granulomatous lesions, as compared to control. The results of the present study suggest that erythromycin attenuates the pulmonary granuloma formation, at least in part, by increasing the production of monocyte chemoattractant protein-1. PMID- 11405513 TI - Reversing acute bronchoconstriction in asthma: the effect of bronchodilator tolerance after treatment with formoterol. AB - Continuous treatment with a short-acting beta2-agonist can lead to reduced bronchodilator responsiveness during acute bronchoconstriction. This study evaluated bronchodilator tolerance to salbutamol following regular treatment with a long-acting beta2-agonist, formoterol. The modifying effect of intravenous corticosteroid was also studied. Ten asthmatic subjects (using inhaled steroids) participated in a randomised, double-blind, placebo-controlled, cross-over study. Formoterol 12 microg b.i.d. or matching placebo was given for 10-14 days with >2 weeks washout. Following each treatment, patients underwent a methacholine challenge to induce a fall in forced expired volume in one second (FEV1) of at least 20%, then salbutamol 100 microg, 100 microg, and 200 microg was inhaled via a spacer at 5 min intervals, with a further 400 microg at 45 min. After a third single-blind formoterol treatment period, hydrocortisone 200 mg was given intravenously prior to salbutamol. Dose-response curves for change in FEV1 with salbutamol were compared using analysis of covariance to take account of methacholine-induced changes in spirometry. Regular formoterol resulted in a significantly lower FEV1 after salbutamol at each time point compared to placebo (p<0.01). The area under the curves (AUCs) for 15 (AUC0-15) and 45 (AUC0-45) min were 28.8% and 29.5% lower following formoterol treatment (p<0.001). Pretreatment with hydrocortisone had no significant modifying effect within 2 h of administration. It is concluded that significant tolerance to the bronchodilator effects of inhaled salbutamol occurs 36 h after stopping the regular administration of formoterol. This bronchodilator tolerance is evident in circumstances of acute bronchconstriction. PMID- 11405514 TI - Effect of oral prednisolone on the bronchoprotective effect of formoterol in patients with persistent asthma. AB - Tolerance to the bronchoprotective effects by long-acting beta2-agonists (LAB) in patients with asthma is not prevented by inhaled corticosteroids (ICS). This study examined whether oral prednisolone can restore the bronchoprotective effects of formoterol in 24 patients with persistent asthma already treated with ICS (at least 800 microg budesonide x day(-1) or equivalent) and LAB, using a parallel-group design. During a 2-week run-in period and during the study, patients used formoterol 12 microg twice daily by Turbuhaler, instead of their own LAB. At baseline and at the end of 7-days treatment with oral placebo or prednisolone (30 mg x day(-1)), provocative concentration of histamine causing a 20% fall in forced expiratory volume in one second (PC20 histamine) was measured on two separate days after randomized single-dose inhalation of placebo (postP) or formoterol (postF). In addition, PC20postF was measured 24 h after starting oral treatment. The protective effect by formoterol at baseline and during treatment was calculated as the difference between the logs of PC20postP and PC20postF. The mean+/-SEM in doubling dose (DD) bronchoprotective effect at baseline was 0.8+/-0.4 DD in the placebo group and 1.0+/-0.4 DD in the prednisolone group. At the end of the treatment period, the protective effect changed to 1.0+/-0.5 DD and 0.8+/-0.6 DD in the placebo and prednisolone treated groups, respectively. This change was not different between the groups (p > 0.4). In conclusion, the bronchoprotective effect by formoterol is not influenced by 1 week prednisolone treatment in patients with asthma who are using regular inhaled corticosteroids and long-acting beta2-agonists. These findings indicate that tolerance to long-acting beta2-agonists cannot be restored by oral steroid therapy. PMID- 11405515 TI - Inhaled corticosteroid therapy reduces the risk of rehospitalization and all cause mortality in elderly asthmatics. AB - Elderly patients with asthma have relatively high rates of hospitalization and mortality. Although inhaled corticosteroids have been shown to improve outcomes among younger patients with asthma, their usefulness in elderly patients has not been established. Therefore, a population-based study of patients 65 yrs of age or older, who have been hospitalized at least once with asthma in Ontario, Canada was conducted to determine the impact of inhaled corticosteroids on rehospitalization for asthma and all-cause mortality rates. Data from the Canadian Institute of Health Information was used to capture all patients 65 yrs of age and older who were hospitalized at least once, with the most responsible diagnosis of asthma in Ontario, Canada between fiscal year 1992 and 1996. This database was then linked with drug claims, physician billing and mortality databases. In total, 6,254 consecutive elderly patients with asthma were identified. Sixty percent of these patients were given at least one prescription for inhaled corticosteroids within 90 days postdischarge from their index hospitalization for asthma. Users of inhaled corticosteroids postdischarge were 29% (95% confidence interval (CI) 20%-38%) less likely to be readmitted to hospital for asthma and 39% (95% CI, 20%-53%) less likely to experience all-cause mortality compared to those who did not receive these drugs postdischarge over a one year follow-up period. These findings suggest that inhaled corticosteroids are beneficial in reducing the risk for rehospitalization and all-cause mortality in elderly patients with asthma who have recently been hospitalized for their disease. PMID- 11405516 TI - Benefit from the inclusion of self-treatment guidelines to a self-management programme for adults with asthma. AB - This study assessed the long-term efficacy of adding self-treatment guidelines to a self-management programme for adults with asthma. In this prospective randomized controlled trial, 245 patients with stable, moderate to severe asthma were included. They were randomized into a self-treatment group (group S) and a control group (group C). Both groups received self-management education. Additionally, group S received self-treatment guidelines based on peak expiratory flow (PEF) and symptoms. Outcome parameters included: asthma symptoms, quality of life, pulmonary function, and exacerbation rate. The 2-yr study was completed by 174 patients. Both groups showed an improvement in the quality of life of 7%. PEF variability decreased by 32% and 29%, and the number of outpatient visits by 25% and 18% in groups S and C, respectively. No significant differences in these parameters were found between the two groups. After 1 yr, patients in both groups perceived better control of asthma and had more self-confidence regarding their asthma. The latter improvements were significantly greater in group S as compared to group C. There were no other differences in outcome parameters between the groups. Individual self-treatment guidelines for exacerbations on top of a general self-management programme does not seem to be of additional benefit in terms of improvements in the clinical outcome of asthma. However, patients in the self-treatment group had better scores in subjective outcome measures such as perceived control of asthma and self-confidence than patients in the control group. PMID- 11405517 TI - Allergic sensitization and diet: ecological analysis in selected European cities. AB - It has been postulated that the prevalence of atopic diseases and their increase over time are associated with regional differences in diet and trends. The results of an ecological correlation study comparing the mean daily intake of selected dietary constituents and the prevalence of allergic sensitization in adults in Europe is presented. Prevalence data from the European Community Respiratory Health Survey (ECRHS) were used. For eight out of 37 ECRHS centres (including 3,872 subjects), comparable dietary data could be obtained. The effect for each dietary constituent was modelled using logistic regression analyses with a term for over dispersion. Inverse associations between the prevalence of allergic sensitization and the energy adjusted intake of fruit were found (odds ratio (OR) = 0.68, p = 0.034). Furthermore OR for energy adjusted intake of vitamin A (OR = 0.73, p = 0.057), vitamin C (OR = 0.83, p = 0.338) and riboflavin (OR = 0.72, p = 0.077) were consistently < 1, but confidence intervals were wider. Daily intake of monounsaturated fatty acids (energy adjusted) was positively associated with sensitization prevalence (OR = 1.59, p = 0.035). These results support the hypothesis that a high intake of monounsaturated fatty acids might promote the development of allergic sensitization. PMID- 11405518 TI - IL-4 fails to regulate in vitro beryllium-induced cytokines in berylliosis. AB - Bronchoalveolar lavage (BAL) cells from patients with chronic beryllium disease (CBD) have been used to evaluate the beryllium-specific immune response and potential immunotherapeutics. Beryllium induces interferon-gamma (IFN-gamma), interleukin-2 (IL-2), tumour necrosis factor-alpha (TNF-alpha), interleukin-6 (IL 6) and interleukin-10 (IL-10) from BAL cells. An antibody to IL-2 and recombinant human (rHu) IL-10 is able to partially suppress the beryllium-stimulated immune response. To obtain BAL cells, bronchoscopy is required, providing risk to the patient and a limited number of cells to study the immune response. As a result, the objectives of the study were to determine 1) whether CBD peripheral blood mononuclear cells (PBMNs) stimulated with beryllium would produce a similar cytokine pattern as BAL cells, and 2) whether this response could be modulated by interleukin-4 (IL-4), an immunomodulatory cytokine. CBD and normal individuals' PBMN and BAL cells were stimulated with and without beryllium sulfate. To modulate this antigen-stimulated response, we added rHu IL-4 to the unstimulated and beryllium-stimulated cells. IFN-gamma, IL-2, TNF-alpha, IL-6 and IL-10 cytokine concentrations were determined from cell supernatants by enzyme-linked immunosorbent assays (ELISA), while IL-4 messenger ribonucleic acid (mRNA) was assessed using polymerase chain reaction (PCR). Beryllium did not stimulate any of these cytokines from normal PBMNs. Increasing levels of IL-6 and TNF-alpha were produced constituitively by CBD PBMNs over time. Compared to the unstimulated CBD PBMNs, beryllium stimulated significant IFN-gamma, TNF-alpha, IL 2, IL-6 and IL-10 production. This response was similar to that stimulated from CBD BAL cells, although of a much lower magnitude. Low levels of IL-4 mRNA were found in CBD and control PBMNs, which were not increased with beryllium stimulation. The beryllium-stimulated cytokine levels were not decreased by the addition of IL-4. IL-4 was unable to downregulate any of these beryllium stimulated cytokines from CBD BAL cells or increase IL-4 mRNA from either CBD PBMN or BAL cells, and thus is an unlikely immunomodulatory agent in CBD. From the data, it was concluded that chronic beryllium disease peripheral blood mononuclear cells provide a model to study the beryllium-stimulated immune response. Interleukin-4's inability to downregulate any of the beryllium stimulated cytokines makes it an unlikely therapeutic candidate in chronic beryllium disease. PMID- 11405519 TI - Gas and dust exposure in underground construction is associated with signs of airway inflammation. AB - Exposure to gases and dust may induce airway inflammation. It was hypothesized that heavy construction workers who had been exposed to dust and gases in underground construction work for 1 yr, would have early signs of upper and lower airway inflammation, as compared to outdoor workers. A study group comprising 29 nonsmoking underground concrete workers (mean +/- SD age 44+/-12 yrs), and a reference group of 26 outdoor concrete workers (39+/-12 yrs) were examined by acoustic rhinometry, nasal and exhaled nitric oxide spirometry and a questionnaire on respiratory symptoms. Exposure measurements were carried out. The underground workers had higher exposure to total and respirable dust, alpha quartz and nitrogen dioxide than the references (p<0.001). The occurrence of respiratory symptoms was higher in the underground workers than in the references (p<0.05). Exhaled nitric oxide (NO) (geometric mean+/-SEM) was higher in the underground workers than in the references (8.4+/-1.09 versus 5.6+/-1.07 parts per billion (ppb), p = 0.001), whereas spirometric values were comparable. The underground workers had smaller nasal cross-sectional area and volume than the references, and more pronounced increases after decongestion (p<0.001). To conclude the exposure in underground construction may cause nasal mucosal swelling and increased levels of exhaled nitric oxide, indicating signs of upper and lower airway inflammation. PMID- 11405520 TI - Higher occurrence of asthma-related symptoms in an urban than a suburban area in adults, but not in children. AB - In young adults, a higher occurrence of asthma-related symptoms was found in an urban than an adjacent suburban area in a survey performed in 1991. The authors now wondered whether such differences could be established in other age groups. The present study (in 1996) included 14,299 subjects, aged 5-75 yrs, of a random sample of the general population in the same two adjacent areas: the centre of Antwerp (Belgium) and its south suburban border. The standardized European Community Respiratory Health Survey (ECRHS) and International Study of Asthma and Allergies in Childhood (ISAAC) questionnaires were used to assess the occurrence of asthma-related symptoms. Higher rates were confirmed in urban compared to suburban Antwerp in adults (20-75 yrs), but no such area differences were found in children (5-8 and 12-15 yrs). Adjustment for a number of recorded risk factors did not seem to affect the area differences in asthma-related symptoms. Comparing the survey results of 1991 and 1996 in 20-44 yr old adults, the findings suggest a slight increase in reported respiratory symptoms in both areas. A higher occurrence of asthma symptoms was observed in the urban than suburban area in adults, but not in children. This might be explained by a progressive effect of long-term exposure to the "urban environment". However, longitudinal studies are necessary to further clarify the factors accounting for these age-related area differences. PMID- 11405521 TI - Ultrafine particles in urban air and respiratory health among adult asthmatics. AB - Airborne particles are associated with adverse health effects and contribute to excess mortality in epidemiological studies. A recent hypothesis proposes that the high numbers of ultrafine (<0.1 microm diameter) particles in ambient air might provoke alveolar inflammation and subsequently cause exacerbations in pre existing cardiopulmonary diseases. To test the hypothesis adult asthmatics were followed with daily peak expiratory flow (PEF) measurements and symptom and medication diaries for six months, while simultaneously monitoring particulate pollution in ambient air. The associations between daily health endpoints of 57 asthmatics and indicators of air pollution were examined by multivariate regression models. Daily mean number concentration of particles, but not particle mass (PM10 (particle mass <10 microm), PM2.5-10, PM2.5, PM1), was negatively associated with daily PEF deviations. The strongest effects were seen for particles in the ultrafine range. However, the effect of ultrafine particles could not definitely be separated from other traffic generated pollutants, namely nitric oxide, nitrogen dioxide and carbon monoxide. No associations were observed with respiratory symptoms or medication use. Particle mass measurements can be strongly influenced by mechanically produced, soil-derived particles, which may not be associated with adverse health effects. Therefore, air quality monitoring should include particle number concentrations, which mainly reflect ultrafine particles. PMID- 11405522 TI - Diet and asthma, allergic rhinoconjunctivitis and atopic eczema symptom prevalence: an ecological analysis of the International Study of Asthma and Allergies in Childhood (ISAAC) data. ISAAC Phase One Study Group. AB - Several studies have suggested that the increasing prevalence of symptoms of asthma, rhinitis and eczema, could be associated with dietary factors. In the present paper, a global analysis of prevalence rates of wheeze, allergic rhinoconjunctivitis and atopic eczema was performed in relation to diet, as defined by national food intake data. Analyses were based on the International Study of Asthma and Allergies in Childhood (ISAAC) data for 6-7 and 13-14 yr old children. Symptoms of wheeze, allergic rhinoconjunctivitis and atopic eczema symptom prevalence were regressed against per capita food intake, and adjusted for gross national product to account for economic development. Dietary data were based on 1995 Food and Agriculture Organisation of the United Nations data for 53 of the 56 countries that took part in ISAAC phase I (1994/1995). The 13-14 year age group showed a consistent pattern of decreases in symptoms of wheeze (current and severe), allergic rhinoconjunctivitis and atopic eczema, associated with increased per capita consumption of calories from cereal and rice, protein from cereals and nuts, starch, as well as vegetables and vegetable nutrients. The video questionnaire data for 13-14 yr olds and the ISAAC data for 6-7 yr olds showed similar patterns for these foods. A consistent inverse relationship was seen between prevalence rates of the three conditions and the intake of starch, cereals, and vegetables. If these findings could be generalised, and if the average daily consumption of these foods increased, it is speculated that an important decrease in symptom prevalence may be achieved. PMID- 11405523 TI - Primary ciliary dyskinesia syndrome associated with abnormal ciliary orientation in infants. AB - Primary ciliary dyskinesia (PCD) syndrome associated with abnormal ciliary orientation but with normal ciliary ultrastructure has been described in adults, but there are no normal ranges for orientation in infants, despite the fact that half of all patients with PCD present in the new-born period. Nasal brush biopsies were obtained from eight infants (three males), mean age 13.1 months, range 7-23, in order to determine ciliary orientation. They had no upper or lower airway disease and normal organ arrangement and were undergoing general anaesthesia for other reasons. Two infants with typical PCD syndrome but normal ultrastructure of individual cilia also had orientation studies. In the eight normal subjects, a mean of 254 central pairs was examined, range 82-453. The mean ciliary orientation was 14.9 degrees, range 12.9-17.5. The two infants with PCD syndrome but normal ultrastructure of individual cilia had ciliary orientation of (Case 1) 44.5 degrees (range 10.6-64.5) in 218 central pairs; and on a second occasion, 28.9 degrees, (range 9.0-47.5) in 259 central pairs; for Case 2, 24.4 degrees, (range 13.1-38.4) in 196 central pairs. The normal range for ciliary orientation is similar in infants to that described in other work in adults. The two cases of phenotypic primary ciliary dyskinesia in the presence of normal ciliary ultrastructure but abnormal ciliary orientation in infants supports the contention that measurement of ciliary orientation should be part of the assessment of ciliary structure and function in cases of possible primary ciliary dyskinesia, in particular when the ultrastructure of individual cilia appear to be normal. PMID- 11405524 TI - Assessment of passive respiratory mechanics in infants: double versus single occlusion? AB - The single breath or occlusion technique (SOT) is widely used to assess passive respiratory mechanics in infants, but depends on various underlying assumptions. Recently, it has been proposed that such measurements could be internally validated by performing two brief airway occlusions during the same expiration. The aim of this study was to evaluate the use of the double occlusion technique (DOT) using a new commercially available program (Jaeger MasterScreen BabyBody Erich Jaeger GmbH, Wurzburg, Germany). Paired measurements of respiratory system compliance (Crs) and resistance (Rrs) using both SOT and DOT were obtained in 18 healthy sedated infants (age range 4-41 weeks, weight 2.7-9.9 kg). There was close agreement between both methods of assessing Crs in all infants, the mean within-subject difference (95% confidence interval (CI)) for DOT-SOT being -0.06 (-0.55- +0.42) mL x kPa(-1) x kg(-1). By contrast, estimates of Rrs,DO were on average 20% lower than those for Rrs,SO, (mean within-subject difference (95% CI) being -0.67 (-1.04- -0.31) kPa x L(-1) x s; p<0.01). The relatively lower values obtained for Rrs,DO may reflect the higher mean lung volume at which it was calculated. Further work is required to investigate the clinical and epidemiological relevance of this new approach, and whether there are any advantages of using both techniques when assessing passive mechanics in infants. PMID- 11405525 TI - Evaluation of a simplified oscillation technique for assessing airway obstruction in sleep apnoea. AB - The forced oscillation technique (FOT) is a noninvasive method that is useful for assessing airway obstruction and for titrating continuous positive airway pressure (CPAP) in patients with sleep apnoea. The aim was to evaluate the routine applicability of a simplified FOT set-up based on recording pressure and flow at the level of the CPAP device, i.e. obviating the need for connecting the transducers to the nasal mask. A correction to account for the tubing and the exhaust port was applied. This simplified FOT was evaluated on nine patients with moderate or severe sleep apnoea during routine CPAP titration. Patient impedance measured by the simplified FOT ([Z]) was compared with actual patient impedance ([Zrs]) measured simultaneously with a reference FOT based on recording pressure and flow at the nasal mask. An excellent agreement was found between [Z] and [Zrs] over the wide range of airway obstruction explored (4.8-72.1 cmH2O x s x L( 1)): [Z] = [Zrs] x 0.86 + 1.3 cmH2O x s x L(-1) (r = 0.99). Moreover, the simplified and the conventional FOT settings detected the same respiratory events during sleep. These results demonstrate that this simplified FOT is applicable for measuring airway obstruction during routine sleep studies in patients with sleep apnoea. PMID- 11405526 TI - Long-term use of mandibular advancement splints for snoring and obstructive sleep apnoea: a questionnaire survey. AB - A mandibular advancement splint (MAS) may be an alternative treatment for snoring and obstructive sleep apnoea (OSA). However, there is little subjective or objective information concerning long-term effectiveness, compliance and side effects. A retrospective questionnaire was used to survey these issues plus patient satisfaction and maintenance requirements in 166 patients who could have worn a mandibular advancement splint for over a year. One-hundred and twenty-six (76%) subjects returned the questionnaire, (84 with OSA, 42 snorers), of whom 69 (55%) reported still using the splint regularly, 47 (37%) every night. The most common reported reasons for stopping use were discomfort (29/ 57; 52%) of nonusers), and poor perceived efficacy (12 subjects). Users reported more daytime symptoms, and they and their partners were more likely to observe improvements with splint use. Side effects were reported by 49 subjects, more commonly in nonusers. Sixty-five of 67 current users and 23 of 41 nonusers reported less snoring with splint use (p = < 0.001). Long-term mandibular advancement splint usage appeared less satisfactory than previously reported, however, splints were considered effective by 97% of current users and even by over half of those who had stopped use. Reasons for stopping use included side effects, social circumstances, dental treatment, as well as lack of perceived efficacy. PMID- 11405527 TI - Proportional positive airway pressure: a new concept to treat obstructive sleep apnoea. AB - Proportional positive airway pressure (PPAP) was designed to optimize airway pressure for the therapy of obstructive sleep apnoea (OSA). In a randomized crossover prospective study, the clinical feasibility of PPAP and its immediate effects on the breathing disorder and sleep in comparison with continuous positive airway pressure (CPAP) was evaluated. Twelve patients requiring CPAP therapy underwent CPAP and PPAP titration in a random order. Obstructive and mixed respiratory events could be completely abolished with both forms of treatment. This efficacy could be achieved at a significantly lower mean mask pressure during PPAP titration (8.45+/-2.42 cmH2O) compared to CPAP (9.96+/-2.7 cmH2O) (p=0.002). The mean minimal arterial oxygen saturation (Sa,O2) (82.8+/ 6.5%) on the diagnostic night increased significantly (p<0.001) to an average Sa,O2 of 93.35+/-1.71% and 93.19+/-2.9% during CPAP and PPAP titration. Total sleep time, slow wave sleep and rapid eye movement (REM) sleep increased significantly by the same amount during both CPAP and PPAP titration (p<0.001), while sleep stage nonrapid eye movement (NREM) 1 and 2 decreased. Six patients preferred the PPAP titration night, four patients did not have a preference, and two patients preferred CPAP. The present data show that proportional positive airway pressure is as effective as continuous positive airway pressure in eliminating obstructive events and has the same immediate effect on sleep. The lower average mask pressure during proportional positive airway pressure implies potential advantages compared to continuous positive airway pressure. Proportional positive airway pressure presents a new effective therapeutic approach to obstructive sleep apnoea. PMID- 11405528 TI - Genetic deficiency of alpha1-PI in mice influences lung responses to bleomycin. AB - It has recently been suggested that proteinase inhibitors modulate the fibrotic response in the lung. This study investigated the development of bleomycin induced pulmonary changes in pallid mice, deficient in serum alpha1-proteinase inhibitor, and with a lower elastase inhibitory capacity, and in congenic C57Bl/6J mice. Male pallid and C57Bl/6J mice received a single intratracheal instillation of either saline or bleomycin. The investigation was carried out by means of biochemical, morphological and morphometrical methods. In both strains, 21 and 72 h after bleomycin, the lungs showed foci of inflammatory cell infiltration associated with emphysema. Fibrosis developed with time after bleomycin. At 14 days fibrosis affected 23.46+/-9.48% (mean +/- SD) and 40.62+/ 13.34% (p < 0.01) of the lungs of C57Bl/6J and pallid mice, respectively. Emphysema affected 3.68+/-3.11% and 12.57+/-4.13% (p<0.01) of lung in C57Bl/6J and pallid mice, respectively. In C57Bl/6J mice bleomycin increased lung hydroxyproline content by 34% and desmosine content by 44% (p < 0.01 for both). In pallid mice these increases were only 21% (p < 0.01) and 6% which may reflect parenchymal loss. Thus, the lung destructive response (emphysema) and the subsequent proliferative reaction (fibrosis) to bleomycin are potentiated in alpha1-proteinase inhibitor deficiency. PMID- 11405529 TI - Diagnostic significance of surfactant proteins A and D in sera from patients with radiation pneumonitis. AB - Radiation pneumonitis (RP) is the most common complication of radiotherapy for thoracic tumours. The aim of this study was to evaluate the significance of pulmonary surfactant proteins (SP)-A and SP-D as new serum markers for RP. Twenty five patients with lung tumour, who had received radiotherapy, were studied. At the completion of radiotherapy, the presence of RP was judged by chest plain radiography and chest high resolution computed tomography (HRCT). RP findings detected on chest plain radiography were seen in only three of 12 patients in whom RP was detected by HRCT. Nevertheless, both SP-A and SP-D concentrations in sera from the patients with RP were significantly higher than those from the 13 patients without RP (p = 0.0065, p = 0.0011, respectively). As with SP-A, ratios of SP-D at the completion, compared to at the initiation (1 week post/pre ratio), were also significantly higher in patients with RP than in patients without RP. When a post/pre ratio > 1.6 was considered positive, the SP-A and SP-D assays showed an 83% and 85% specificity, respectively. In conclusion, serum assays of surfactant proteins A and D may be of diagnostic value for detection of radiation pneumonitis, even when the radiographic change is faint. PMID- 11405530 TI - Effects of mechanical ventilation of isolated mouse lungs on surfactant and inflammatory cytokines. AB - Mechanical ventilation of the lung is an essential but potentially harmful therapeutic intervention for patients with acute respiratory distress syndrome. The objective of the current study was to establish and characterize an isolated mouse lung model to study the harmful effects of mechanical ventilation. Lungs were isolated from BalbC mice and randomized to either a nonventilated group, a conventionally ventilated group (tidal volume 7 mL x kg(-1), 4 cm positive endexpiratory pressure (PEEP)) or an injuriously ventilated group (20 mL x kg( 1), 0 cm PEEP). Lungs were subsequently analysed for lung compliance, morphology, surfactant composition and inflammatory cytokines. Injurious ventilation resulted in significant lung dysfunction, which was associated with a significant increase in pulmonary surfactant, and surfactant small aggregates compared to the other two groups. Injurious ventilation also led to a significantly increased concentration of interleukin-6 and tumour necrosis factor-a in the lavage. It was concluded that the injurious effects of mechanical ventilation can effectively be studied in isolated mouse lung, which offers the potential of studying genetically altered animals. It was also concluded that in this model, the lung injury is, in part, mediated by the surfactant system and the release of inflammatory mediators. PMID- 11405531 TI - Airway resistance measured by the interrupter technique: expiration or inspiration, mean or median? AB - The measurement of airway resistance by the interrupter technique (Rint) needs standardization. Should measurements be made be during the expiratory or inspiratory phase of tidal breathing? In reported studies, the measurement of Rint has been calculated as the median or mean of a small number of values, is there an important difference? Subjects were 2.5-5.0 yrs (median 4.0 yrs) who had previous respiratory symptoms. The Rint in expiration (RintE) and inspiration (RintI) pre and postsalbutamol, the coefficient of variation (CV) of values contributing to measurements, and bronchodilator responsiveness(BDR) in both phases were compared. Measurements using median and mean were compared. RintE was higher than RintI by 4% (p < 0.01). The CV of values making up RintE and RintI, and BDR measured in expiration and inspiration were similar. The median difference between means and medians of values making up measurements was 0.6% (range -6-11%). RintE has been shown to be consistently greater then RintI but the difference in this study is small. It is suggested that one or the other is chosen as the standard. In the present data the mean of a set of values contributing to a measurement was not significantly different from the median. However, the use of the median has been recommended since it is less affected by possible outlying values such as might be included by fully automated equipment. PMID- 11405532 TI - New anti-asthma therapies: suppression of the effect of interleukin (IL)-4 and IL 5. AB - Asthma is currently defined as a chronic inflammatory disorder of the airways. The central role of allergen-specific Th2 cells in the regulation of this mucosal airway inflammation has been highlighted. Hence, there is large interest in the therapeutic potential of an anti-Th2 cell approach. One of the strategies which has been developed, is to inhibit the effect of interleukin (IL)-4 or IL-5, two main Th2 cell derived cytokines. Interleukin-4 is pivotal in the pathogenesis of allergic disorders through its wide range of effects. An important observation, especially during secondary antigen exposure, is the possible redundancy with IL 13. Both cytokines share common elements in their receptor and intracellular signalling pathway. As a result, compounds can be developed that selectively inhibit the effect of either IL-4 or IL-13, or alternatively, by interfering with the common pathway, inhibit the effect of both cytokines. Eosinophils are generally seen as a particularly harmful element in the allergic inflammation. The importance of IL-5 on eosinophil biology has clearly been established. Conversely, in man, the biological effects of IL-5 are largely limited to eosinophil function. Therefore, IL-5 antagonists offer the unique opportunity of selectively neutralizing the effect of eosinophils. Several strategies have now been developed that successfully inhibit the biological effect of interleukin-4 or interleukin-5. Some of these compounds have proven to be biologically active in man. The challenge now is to establish their therapeutic role in asthma. PMID- 11405533 TI - Imaging guided thoracic interventions. AB - Interventional Radiology is a technique based medical specialty, using all available imaging modalities (fluoroscopy, ultrasound, computed tomography, magnetic resonance, angiography) for guidance of interventional techniques for diagnostic or therapeutic purposes. Actual, percutaneous transthoracic needle biopsy includes core needle biopsy besides fine needle aspiration. Any pleural, pulmonary or mediastinal fluid or gas collection is amenable to percutaneous pulmonary catheter drainage. Treatment of haemoptysis of the bronchial artery or pulmonary artery origin, transcatheter embolization of pulmonary arteriovenous malformations and pseudoaneurysms, angioplasty and stenting of the superior vena caval system and percutaneous foreign body retrieval are well established routine procedures, precluding unnecessary surgery. These techniques are safe and effective in experienced hands. Computed tomography is helpful in pre- and postoperative imaging of patients being considered for endobronchial stenting. Many procedures can be performed on an outpatient basis, thus increasing the cost effectiveness of radiologically guided interventions in the thorax. PMID- 11405534 TI - The bias flow nitrogen washout technique for measuring the functional residual capacity in infants. ERS/ATS Task Force on Standards for Infant Respiratory Function Testing. AB - The functional residual capacity (FRC) is the most commonly measured static lung volume in infants. It is important for interpreting volume-dependent pulmonary mechanics, e.g. airway resistance, and defining normal lung growth. The bias flow nitrogen washout technique is widely used for measuring FRC because the dead space and circuit resistance are low, making it suitable for small or sick infants. Moreover, data acquisition and calculation are easily programmed for a personal computer. The aim of this paper is to provide recommendations pertaining to equipment requirements, study procedures and reporting of data for functional residual capacity measurements. While measuring the functional residual capacity is regarded as physiologically and clinically important, the accuracy of the measurement is undoubtedly equally important. Hence, the paper also emphasizes factors influencing the accuracy of functional residual capacity measurements independent of equipment requirements. These recommendations represent the "State of the Art" in 2000. PMID- 11405535 TI - M. tuberculosis: immunology and vaccination. AB - Tuberculosis is increasing. Current treatment regimens require at least 6 months, because latent or stationary phase organisms are difficult to kill. Such regimens do not achieve full compliance, and "directly observed therapy short course" (DOTS) is having less impact than expected. This worrying situation is aggravated by coinfection with human immunodeficiency virus (HIV), and by the increase in drug-resistant strains. We need new insights that lead to more rapid therapies and immunotherapies, and more reliable vaccines. Recent insights have come from: understanding of the relationship between Mycobacterium tuberculosis and macrophages; the multiple T cell types that recognise mycobacterial peptides, lipids and glycolipids; the critical role of interferon-gamma (IFNgamma) and interleukin-12 (IL-12) in human mycobacterial infection revealed by genetically defective children; quantitation of the presence and importance of Th2 lymphocyte activation in human tuberculosis; the role of local conversion of inactive cortisone to active cortisol in the lesions; the recognition that some effective prophylactic vaccines also work as immumotherapeutics whereas others do not. In the longer term the recent sequencing of the M. tuberculosis genome will lead to further advances. In the short term, effective immunotherapy remains the most accessible breakthrough in the management of tuberculosis. The types of practical advance that will result from sequencing the genome are discussed speculatively, but cannot yet be predicted with certainty. PMID- 11405536 TI - A thousand years of pulmonary medicine: good news and bad. PMID- 11405537 TI - Tracheobronchial stenosis in Keutel syndrome. AB - In 1971 Keutel et al. described a new syndrome in two siblings presenting with peripheral pulmonary stenoses, brachytelephalangism, neural hearing loss and abnormal cartilage calcification. Recent investigations provided evidence that mutations in the gene encoding the human matrix GLA protein cause Keutel syndrome. With these new insights in the disease the symptomatology of Keutel syndrome was reassessed. The follow-up of the two siblings was studied by clinical and post mortem examination. As a new feature of Keutel syndrome tracheobronchial stenosis and concentric calcification of pulmonary, coronary, hepatic, renal, meningeal and cerebral arteries were described. Complementary to the results in molecular genetics the symptomatology of Keutel syndrome could be revised by clinical and post mortem examination. PMID- 11405538 TI - X-linked agammaglobulinaemia and squamous lung cancer. AB - A 32yr-old nonsmoking male, diagnosed as having X-linked agamma-globulinemia, presented with fever, cough with purulent sputum, a very intense back pain and a mass of 10 centimetres in lower left lobe. Diagnostic evaluation revealed a squamous cell carcinoma with very aggressive metastases at L3. Malignancies are the second leading cause of death in children and adults with congenital immunodeficiency disorders, mostly non-Hodgkin lymphomas and gastric and colon adenocarcinomas, but this is the first report of lung cancer in a patient with X linked agammaglobulinemia. Lung cancer incidence has been reported to be higher in patients with other diseases of the lung, however, there is no clear evidence of the role of bronchiectasis in developing lung cancer. It is possible that a longer survival for patients with X-LA recently diagnosed, and an association of chronic bronchial infection, could favour the development of pulmonary neoplasm. PMID- 11405539 TI - The risk of future cardiovascular diseases in the patients with OSAS is dependently or independently associated with obstructive sleep apnoea. PMID- 11405540 TI - Does theophylline really improve acute mountain sickness? PMID- 11405541 TI - Heart rate as a cardiovascular risk factor: do women differ from men? AB - Considerable progress has been made in our understanding of the role of high heart rate in determining cardiovascular morbidity and mortality. However, whether the association between fast heart rate and cardiovascular disease is equally strong in males and females is still a matter for debate. In most studies, the predictive value of tachycardia for all-cause mortality has been found to be weaker in women than in men, and in some studies no association between heart rate and cardiovascular mortality was observed. In particular, high heart rate appeared to be a weak predictor of death from coronary heart disease in the female gender. Multiple mechanisms by which sympathetic overactivity could cause hypertension and the metabolic syndrome of insulin resistance have been documented. Recent results obtained at the Ann Arbor laboratory from the analysis of four populations indicate that these mechanisms are operative mostly in males in whom tachycardia reflects a heightened sympathetic tone. In women, fast heart rate would merely represent the extreme of a normal distribution. However, tachycardia can also have a direct impact on the arterial wall, as demonstrated in laboratory studies, and can favour the occurrence of cardiac arrhythmias. The impact of these mechanisms may be similar in men and women and could explain why a high heart rate has been found to have a detrimental effect also in the female gender. Pharmacological reduction of high heart rate is an additional desirable goal of therapy in several clinical conditions such as hypertension, myocardial infarction and congestive heart failure. Although a greater effect is expected in men, cardiac slowing could counteract the detrimental haemodynamic effect of tachycardia also in women. PMID- 11405542 TI - Telemedicine framework and applications in dermatology and ophthalmology. AB - Medicine has to balance between the advantages and costs of new technology. As the significance of technical aids has increased, medicine has become more tightly bound to technology. Telemedicine is one of the fastest developing fields, as its development is connected to the development of telecommunication and information technology. Technology sets the ultimate restrictions to telemedicine. However, most challenges are nontechnical. Fast development makes it difficult to perform generalizable studies on the field, and the lack of practical, applicable standardization hinders telemedical system design. The cost of technology is quickly falling compared with the cost of human labour. Because of these factors the efficient use of telemedicine requires strategic decisions at the level of the organization as well as more research concerning the effects of telemedicine on medical practice. This article describes the telemedical frame of reference by using dermatology and ophthalmology as examples. PMID- 11405543 TI - Therapeutic potential of the intestinotropic hormone, glucagon-like peptide-2. AB - Glucagon-like peptide-2 (GLP-2) is a newly discovered growth factor that has been demonstrated to enhance intestinal growth and function in normal rodents and to prevent damage and facilitate intestinal repair in various animal models of intestinal insufficiency. A recent study has demonstrated that GLP-2 also acts as an intestinotropin in humans with short-bowel syndrome. The high degree of specificity of GLP-2 for induction of intestinal growth, without affecting growth of other peripheral tissues, is determined by the highly localized expression of the GLP-2 receptor in the intestinal epithelium. In this article, we review the regulation of GLP-2 in physiology, from synthesis to metabolism, with a particular emphasis on potential targets in this pathway for therapeutic manipulation of GLP-2 actions. We also discuss the various animal models of intestinal insufficiency that have been used to demonstrate the therapeutic potential of this intestinotropic hormone, including short bowel, intestinal atrophy, enteritis and colitis. The results of these studies indicate that GLP-2 is a promising therapeutic agent for the treatment of various forms of intestinal insufficiency in humans. PMID- 11405544 TI - Ventricular interaction: from bench to bedside. AB - Decreased right ventricle (RV) output results in decreased left ventricle end diastolic volume (LVEDV) and output by series interaction. Direct ventricular interaction may also have a major effect on LV function. Thus, decreased LVEDV caused by reduced RV output may be further reduced by a leftward septal shift and pericardial constraint. This has been shown to be true in acute and chronic pulmonary hypertension and is now also apparent in severe congestive heart failure. The use of intracavitary LV end-diastolic pressure (LVEDP) to assess LVEDV is inappropriate if pressure surrounding the LV is increased: the surrounding pressure should be subtracted from LVEDP to calculate the effective distending (transmural) pressure which governs preload. If the surrounding pressure increases more than LVEDP, transmural LVEDP and LVEDV will decrease despite the increased LVEDP. Thus, the use of filling pressure to reflect changes in LVEDV has led to erroneous conclusions regarding changes in myocardial compliance and contractility. It is now clear that volume loading may reduce LVEDV and stroke work in pulmonary embolism, chronic lung disease and severe congestive heart failure despite increased LVEDP. The decreased stroke work is a result of reduced LV preload, not decreased contractility as would be suggested if filling pressure is used to reflect preload. PMID- 11405545 TI - Molecular genetics and the epidemiology of bipolar disorder. AB - The methodologies of epidemiology and molecular genetics are complementary approaches to identifying risk factors in bipolar disorder. Genetic linkage studies have revealed several chromosomal loci likely to contain genes that increase the risk of bipolar disorder, but major uncertainties remain about the mode of inheritance of the condition and the definition of the phenotype. Epidemiological findings have contributed to both these areas and have led to new hypotheses about causation. For example, the analysis of variability of age at onset of bipolar disorder led to studies of anticipation and a possible role of dynamic DNA repeat sequence mutations. Future epidemiological studies that aim to identify risk factors for bipolar disorder at the population level will be able to measure the interactions of genome sequence variation with other risk factors in the domain of demography, childhood experiences, exposure to adversity and availability of social support. PMID- 11405546 TI - Candidate gene studies of bipolar disorder. AB - Genetic factors undoubtedly play an important role in determining vulnerability to bipolar disorder but the task of finding susceptibility genes is not trivial. Candidate gene studies, usually employing the association approach, offer the potential to discover the genes of relatively modest effect size that are expected for a complex genetic disorder. Candidate gene approaches depend crucially on our current understanding of disease pathophysiology, and attention has consequently been focussed on a limited range of neurotransmitter systems implicated by the action of drug treatments. Despite no unequivocal, consistently replicated findings, a number of intriguing results have emerged in the literature, both for bipolar disorder in general and for subtypes such as bipolar affective puerperal psychosis and rapid cycling bipolar illness. Genes of particular current interest include those encoding the serotonin transporter, monoamine oxidase A (MAOA) and catechol-O-methyl transferase (COMT). As susceptibility genes are found and knowledge of aetiology advanced it is likely that many more candidate genes in novel biological systems will attract attention. PMID- 11405547 TI - Genome scans for susceptibility genes in bipolar affective disorder. AB - A genome-wide scan for genetic linkage can suggest fresh insights into disease aetiology. However, in the case of complex disorders such as bipolar affective disorder (BPAD), the results of genome-wide scans must be interpreted with caution. We review 10 published and 10 in-progress genome scans of BPAD, encompassing 3536 affected individuals in 1119 pedigrees. We find that ascertainment methods vary widely, with no two studies using identical methods. Sample sizes and marker densities have generally been well below what is now considered adequate, but several in-progress studies are using larger samples and more closely spaced markers. Few findings reach the 'suggestive' threshold, and fewer still reach the 'significant' threshold at genome-wide levels of significance. Strategies for pooling samples or subjecting findings in different samples to meta-analysis are being developed, but differences in ascertainment methods may have a large impact on the uniformity of different samples and hamper efforts at combining data or findings. There is also a need for methods that help define more genetically homogeneous phenotypes, take into account interactions between multiple susceptibility loci, and accommodate additional complexity (eg parent-of-origin effects) in the search for linkage. PMID- 11405548 TI - Convergent functional genomics: application to bipolar disorder. AB - The recent development of microarray technologies has made possible the simultaneous measurement of mRNA levels for thousands of genes and a new genomic method termed gene expression profiling. The application of this approach to animal models or post-mortem tissue provides a powerful tool for the discovery of novel genes involved in psychiatric disorders. This approach has strengths that are complementary to those of another genomic method for gene discovery, positional cloning. Microarray technologies and their application to post-mortem tissue and animal models of bipolar disorder are reviewed. A novel approach termed convergent functional genomics, which integrates gene profiling and positional cloning in order to rapidly identify candidate disease genes, is also described. PMID- 11405549 TI - Lithium-related genetics of bipolar disorder. AB - Lithium is a potent prophylactic medication and mood stabilizer in bipolar disorder. However, clinical outcome is variable, and its therapeutic effect manifests after a period of chronic treatment, implying a progressive and complex biological response process. Signal transduction systems known to be perturbed by lithium involve phosphoinositide (PI) turnover, activation of the Wnt pathway via inhibition of glycogen synthase kinase-3beta (GSK-3beta), and a growth factor induced, Akt-mediated signalling that promotes cell survival. These pathways, acting in synergy, probably prompt the amplification of lithium signal causing such immense impact on the neuronal network. The sequencing of the human genome presents an unparallelled opportunity to uncover the full molecular repertoire involved in lithium action. Interrogation of high-resolution expression microarrays and protein profiles represents a strategy that should help accomplish this goal. A recent microarray analysis on lithium-treated versus untreated PC12 cells identified multiple differentially altered transcripts. Lithium-perturbed genes, particularly those that map to susceptibility regions, could be candidate risk-conferring factors for mood disorders. Transcript and protein profiling in patients could reveal a lithium fingerprint for responsiveness or nonresponsiveness, and a signature motif that may be diagnostic of a specific phenotype. Similarly, lithium-sensitive gene products could provide a new generation of pharmacological targets. PMID- 11405551 TI - Improvement of nutrient absorption may enhance systemic oxidative stress in cystic fibrosis patients. AB - BACKGROUND: The life expectancy of patients with cystic fibrosis (CF) is largely dependent on the pulmonary disease severity and progress. Malnutrition may be an important complicating factor in active and chronic lung disease. AIMS: The focus of this study was to investigate several inflammatory markers in pancreatic insufficient CF patients with different enzyme treatment regimens. METHODS: CF patients with pancreatic insufficiency were examined at a time of symptomatic exacerbation of their lung disease. Group A (n = 11) regularly received microspheric enzymes. Group B (n = 8) were treated with enzymes during the hospitalization period only and demonstrated the presence of malnutrition. Inflammatory markers in the sputa (neutrophil elastase activity, interleukin-8 and tumour necrosis factor-alpha levels) and in the peripheral blood (plasma malondialdehyde (MDA), lymphocyte response to PHA, and the cell sensitivity to steroid suppression) have been investigated. RESULTS: During acute lung exacerbation, group B demonstrated reduced levels of lymphocyte proliferation. This parameter was normalized after combined antibiotic and pancreatic enzyme therapy. Simultaneously, plasma MDA in group B markedly increased following treatment. For this group, a significant positive linear association between values of plasma MDA and lymphocyte proliferation has been observed. For group A, neither the same correlation nor changes in MDA levels and lymphocyte proliferation have been found. CONCLUSIONS: Our data indicate that acute lung exacerbation in malnourished CF patients may be associated with alteration in T lymphocyte activity. Adequate therapy normalizes lymphocyte function but results in systemic oxidative stress. PMID- 11405550 TI - Anti-inflammatory cytokines in asthma and allergy: interleukin-10, interleukin 12, interferon-gamma. AB - Interleukin-10 (IL-10) is a cytokine derived from CD4+ T-helper type 2 (T(H2)) cells identified as a suppressor of cytokines from T-helper type 1(T(H1)) cells. Interleukin-12 (IL-12) is produced by B cells, macrophages and dendritic cells, and primarily regulates T(H1) cell differentiation, while suppressing the expansion of T(H2) cell clones. Interferon-gamma (IFN-gamma) is a product of T(H1) cells and exerts inhibitory effects on T(H2) cell differentiation. These cytokines have been implicated in the pathogenesis of asthma and allergies. In this context, IL-12 and IFN-gamma production in asthma have been found to be decreased, and this may reduce their capacity to inhibit IgE synthesis and allergic inflammation. IL-10 is a potent inhibitor of monocyte/macrophage function, suppressing the production of many pro-inflammatory cytokines. A relative underproduction of IL-10 from alveolar macrophages of atopic asthmatics has been reported. Therapeutic modulation of T(H1)/T(H2) imbalance in asthma and allergy by mycobacterial vaccine, specific immunotherapy and cytoline-guanosine dinucleotide motif may lead to increases in IL-12 and IFN-gamma production. Stimulation of IL-10 production by antigen-specific T-cells during immunotherapy may lead to anergy through inhibition of CD28-costimulatory molecule signalling by IL-10s anti-inflammatory effect on basophils, mast cells and eosinophils. PMID- 11405552 TI - Increased urinary nitrite, a marker of nitric oxide, in active inflammatory bowel disease. AB - BACKGROUND: Nitric oxide (NO) production is increased in inflammatory bowel disease (IBD), and measurement of NO metabolites may be useful for monitoring disease activity. AIMS AND OBJECTIVES: To characterise urinary nitrite levels, a stable metabolite of NO, in IBD and to evaluate its potential as a marker of disease activity. METHODS: Twelve-hour urinary nitrites were measured by the microplate assay method in 46 patients with IBD (active; n = 32). Urinary samples from 16 healthy individuals served as controls. RESULTS: Increased levels of urinary nitrites were found in patients with active IBD compared with those with inactive IBD. Twenty-eight out of 32 patients (87.5%) with active IBD had detectable levels of nitrite in their urine as compared with 2/14 (14.3%) patients with inactive IBD. None of the 16 healthy controls had detectable urinary nitrite. Twelve-hour urinary nitrite in active compared with inactive IBD: 5 0.7 versus 0.1+/-0.04 micromol (P < 0.05). There was good correlation between urinary nitrite and some markers of disease activity in IBD such as C reactive protein and microalbuminuria but not with erythrocyte sedimentation rate. CONCLUSIONS: Increased levels of nitrite were detected in urine of patients with active IBD, consistent with increased NO synthesis. This simple assay may be exploited as a potential marker of disease activity in IBD. PMID- 11405553 TI - The possible anti-inflammatory role of circulating human leukocyte antigen levels in women with endometriosis after treatment with danazol and leuprorelin acetate depot. AB - BACKGROUND: Endometriosis is defined as an inflammatory condition of the female reproductive tract, a state often associated with infertility and miscarriage. Many exogenously administered factors (treatments) control the disease via as yet unknown pathways. Possible candidate molecules involved in these mechanisms could be the serum-soluble human leukocyte antigens (sHIA) that have been detected in a variety of human body fluids and that are associated with several diseases. AIMS: We here examine how danazol and leuprorelin acetate depot treatments exert their anti-inflammatory action. It is plausible that subtle alterations mediated by these treatments and in relation to sHLA may explain the pathophysiology of endometriosis and provide insights towards new therapeutic protocols. METHODS: Indirect enzyme-linked immunosorbent assay (ELISA), using specific monoclonal antibodies, determined serum-soluble class-I and class-II HLA levels. ELISA readings from treated women were compared with normal healthy subjects. RESULTS: Serum-soluble class-I and class-II HLA levels are statistically significantly lower (P < 0.001) in women with endometriosis than in the control groups. However, danazol but not leuprorelin acetate depot administration augments soluble HLA class I and class II (P < 0.01 and P < 0.001, respectively) to normal levels during the treatment period, an increase that may account for the anti inflammatory effect and the remission observed. CONCLUSIONS: It is shown that one of the underlying causes of endometriosis may be the lack of both circulating class-I and class-II antigen levels. Danazol administration acts via an induced release of these antigens, whose presence correlates with the degree of the inflammatory alleviation obtained. We thus provide evidence that the inflammatory state of the disease appears to be associated with soluble HLA levels because, 3 months after ceasing therapy, the circulating antigens in the serum return to the same levels that correspond to the pathological condition. PMID- 11405554 TI - Release of tumor necrosis factor-alpha and prostanoids in whole blood cultures after in vivo exposure to low-dose aspirin. AB - BACKGROUND: The preventive effect of low-dose aspirin in cardiovascular disease is generally attributed to its antiplatelet action caused by differential inhibition of platelet cyclooxygenase-1. However, there is evidence that aspirin also affects release of inflammatory cytokines, including tumor necrosis factor alpha (TNF-alpha). It is not known whether this is caused by direct action on the cytokine pathway or indirectly through cyclooxygenase inhibition and altered prostanoid synthesis, or both. METHODS: We assessed the capacity of lipopolysaccharide-activated leukocytes in whole blood cultures of eight healthy subjects following a single oral dose of 80 mg aspirin to release TNF-alpha, prostanoid E2 (PGE2) and prostanoid I2 (PGI2), and thromboxane A2 (TXA2). TNF alpha and prostanoids were determined by enzyme-linked immunoassays. RESULTS: In seven subjects, TNF-alpha release in blood cultures decreased 24h after intake of aspirin. The effect of aspirin on prostanoid release was assessed in three individuals: PGE2 increased in all subjects, PGI2 increased in two and remained unchanged in one, and TXA2 was reduced in two and unchanged in one individual The presence of DFU, a specific inhibitor of cyclooxygenase 2, did not affect the reduction of TNF-alpha release by aspirin, but abolished prostanoid production in all three individuals. CONCLUSION: The capacity of activated leukocytes to release TNF-alpha is reduced by ingestion of low-dose aspirin, independent of changes in prostanoid biosynthesis. PMID- 11405555 TI - Syntheses of two deacetyl-thymosin beta4 analogs and their anti-inflammatory effect on carrageenin-induced edema in the mouse paw. AB - Two [Met(0)6]deacetyl-thymosin beta4 analogs containing Phe(4F) or Tyr(Me) at position 12 were synthesized by the manual solid-phase method, and their anti inflammatory effect on carrageenin-induced edema in the mouse paw was studied. Fluorination of the para-position of Phe12 resulted in a marked antiinflammatory effect on carrageenin-induced edema in the mouse paw compared with that of our synthetic [Met(0)6]deacetyl-thymosin beta4, but the other analog, [Met(0)6, Tyr(Me)12]deacetyl-thymosin beta4, showed a marked reduction of the anti inflammatory effect. PMID- 11405556 TI - Administration of isoferulic acid improved the survival rate of lethal influenza virus pneumonia in mice. AB - BACKGROUND: Isoferulic acid (IFA) is a main active ingredient of the rhizoma of Cimicifuga beracleifolia, which is used frequently in Japanese traditional medicine as an anti-inflammatory drug. It has been revealed that IFA inhibits the production of macrophage inflammatory protein-2 (MIP-2), which is a murine counterpart of the chemokine family that may contribute to the pathogenesis of inflammatory diseases through the chemotactic activity for inflammatory and immune effector cells. AIM OF THE STUDY: In this study, we investigated the therapeutic effect of IFA on the progression of lethal influenza virus pneumonia in mice by comparison with that of dexamethasone (DX), a potent inhibitor for various inflammatory cytokines including MIP-2. METHODS: Mice were infected by intranasal inoculation of influenza virus under ether anesthesia. The IFA or DX was given by oral administration once daily for 4 days after infection. After infection, the survival rate and the change in body weight were daily monitored. RESULTS: IFA administration markedly improved the survival rate and body weight loss of influenza virus-infected mice in a suitable dose range (0.5 mg/day). However, DX administration did not show a beneficial effect at any dose. CONCLUSION: These data suggested that IFA is a novel tool not only for the intervention therapy, but also for the studies on the pathogenesis of influenza virus-induced pneumonia. PMID- 11405557 TI - Hepatitis C: issues from the Southeast Asian perspectives. PMID- 11405558 TI - Seroprevalence of anti-HCV in an urban child population: a preliminary study from Kuala Lumpur. AB - A pilot study to determine the seroprevalence of anti-HCV among children from Kuala Lumpur, Malaysia, was conducted using microparticle enzyme immunoassay. Serum samples were obtained randomly from children, aged between one to 16 years of age, admitted to the paediatric unit of University of Malaya Medical Centre, Kuala Lumpur for various medical reasons. Of the 179 samples assayed, only one was positive, giving the prevalence rate of 0.6%. It is reasonable to conclude that the seroprevalence of anti-HCV among children from Kuala Lumpur is low, less than 1%. PMID- 11405559 TI - Pulmonary artery diameters in premature infants: normal ranges. AB - The aim of this study is to establish the norms for pulmonary arterial diameters in the premature infants. One hundred and thirty cross-sectional echocardiograms were performed on 62 premature neonates (23.4 weeks to 36 weeks gestation) in the Neonatal Intensive Care Unit. Except for small atrial septal defects/patent foremen ovale (< or =3 mm) or patent ductus arteriosus (PDA), babies with structural heart defects were excluded. The weight at echocardiography ranges from 470 grams to 2,445 grams, with a mean of 1,157 grams. The diameter of the pulmonary annulus (PA), left pulmonary artery (LPA) and right pulmonary artery (RPA) were measured at peak systole at predetermined sites. Sizes of the atrial septal defect and PDA were also measured, if present. There was no difference in the diameter between the left and right pulmonary arteries (p=0.254, paired t test) in the same patient. After controlling for weight, the mean diameters of the LPA and RPA were larger in patients with PDA (p=0.002) compared to those without PDA (p=0.002), while their pulmonary annulus were comparable in size (p=0.691). Between the gestational ages of 23 and 36 weeks, the diameter of PA, LPA and RPA correlated linearly with weight (Pearson R = 0.84, 0.82, 0.65 and 0.71, respectively; p<0.0005). Prediction graphs and regression equations are given. These normal ranges can be used for assessment of pulmonary artery diameters in premature neonates. PMID- 11405560 TI - Study on antimicrobial susceptibility of bacteria causing neonatal infections: a 12 year study (1987-1998). AB - OBJECTIVE: The method of Manual of Clinical Microbiology was used to identify bacteria. We investigated the epidemiological characteristics of bacterial agents and their antimicrobial susceptibility as empirical treatment for neonatal infections. Disk diffusion tests were also done for antimicrobial susceptibility. RESULTS: From January 1987 to December 1998, 2,244 bacterial strains were isolated in our neonatal ward. The first three predominant species were Staphylococcus epidermidis (23.9%), Staphylococcus saprophyticus (19.9%) and Escherichia coli (12.6%) in group I (infections acquired outside of hospital). Escherichia coli, Klebsiella and Pseudomonas aeruginosa accounted for 18%, 15.2% and 12.6% respectively in group II (nosocomial infections). The sensitivity rates of those antimicrobials that are seldom used for newborns were found to be higher, while the resistant rates of the commonly used antimicrobial drugs have increased significantly. The resistant rates of bacterial isolate from group II to antimicrobial agents including penicillin and ampicillin were significantly higher than those isolated from group I (p<0.05)The sensitivity rate was 82.2% (717/833) by using amikacin only, when combined with penicillin, rose to 89%(741/833). CONCLUSIONS: Gram-negative bacteria were mainly responsible for nosocomial infections of neonates in our hospital. Infections acquired outside the hospital were mainly caused by Gram-positive bacteria. Nosocomial pathogens produced drug resistance easily. Combination of amikacin and penicillin can be recommended as the initial antibiotics for treatment of neonatal infections. PMID- 11405561 TI - Stigma of mental illness. AB - BACKGROUND: Psychiatric patients carry the additional burden of stigma. METHODS: The views of 300 psychiatric out-patients and day-patients and 100 mental health workers concerning stigma were sought. The control group comprised 50 cardiac out patients. RESULTS: A fair proportion of patients with schizophrenia or depression perceived that stigma had a negative effect on their self-esteem, relationships and job opportunities. The majority felt a need for an increase in public awareness of mental illness. In contrast, the cardiac patients reported very little stigmatization. CONCLUSIONS: The diagnostic label of mental illness may render the person vulnerable to stigmatization. Possible causes of stigma and ways of reducing stigma are discussed. PMID- 11405562 TI - Gynaecomastia and the herbal tonic "Dong Quai". AB - We present a case of a man who developed gynaecomastia after ingestion of "Dong Quai" pills. "Dong Quai" is the Chinese name for the herb Angelica polymorpha var. sinensis which is widely used as a panacea for gynaecological problems, and it is also proclaimed as an invigorating tonic for both women and men. The pharmacological effects of "Dong Quai" are likely related to the phytoestrogen that it contains. This report highlights the potential adverse effects associated with its consumption in the male, especially for the processed "Dong Quai" pills which may contain significantly higher levels of phytoestrogen than its original herbal product. PMID- 11405563 TI - Acute respiratory failure in melioidosis. AB - BACKGROUND: In melioidosis caused by Burkholderia pseudomallei, although every organ in the body may be involved, the highest mortality of 73% occurs when the respiratory system is affected. These patients invariably die of acute respiratory failure. Most of them also have underlying predisposing factors like diabetes mellitus. AIM OF STUDY: A retrospective study of six such cases was carried out in order to elicit the possible causes and mechanisms of acute respiratory failure in patients with melioidosis. METHOD: Patients' records were reviewed for demographic, clinical, laboratory, radiological and histopathological data. RESULTS: The rapidity of onset of respiratory failure was remarkable and was accompanied by relentless hypoxaemia that was refractory to treatment despite the application of high positive end expiratory pressure and other supportive measures. All had bilateral opacities on frontal chest radiographs, focal and diffuse necrotizing pneumonia and presence of hyaline membranes in lung tissues seen histologically, supporting the accepted criteria for ALI/ARDS. CONCLUSION: Patients with sepsis due to B. pseudomallei develop ALI/ARDS very rapidly resulting in high mortality rates. Possible mechanisms involved are discussed. Awareness of the disease in endemic areas, the development of rapid diagnostic methods and appropriate management procedures are urgently needed for the prevention of ARDS and subsequent reduction in mortality in such cases. PMID- 11405564 TI - Peripartal cardiomyopathy: problems in diagnosis and management. AB - Peripartal cardiomyopathy is a rare form of heart disease in pregnancy with an unpredictable outcome. We describe one patient who presented in a decompensated state who was successfully managed with medical antifailure treatment. The etiology, management and future obstetric outcome are discussed. PMID- 11405565 TI - Wernicke's encephalopathy in patients with hyperemesis gravidarum. AB - Our two patients presented with Wernicke's Encephalopathy (WE) resulting from prolonged hyperemesis gravidarum. This is an unusual cause of WE, a potentially fatal medical emergency due to thiamine deficiency. We discuss the clinical settings, presentation, diagnosis, neurophysiological and radiological findings, treatment and outcome of WE in these two cases and the neuropathologic correlation of this condition. We stress upon the importance of early diagnosis and prompt treatment of WE. PMID- 11405566 TI - Post-splenectomy multiple pancreatic pseudocysts. AB - A patient was admitted for breathlessness associated with post-splenectomy multiple pseudocysts and succumbed after internal drainage of the pseudocyst. Although the occurrence of pseudocyst following splenectomy is uncommon, failure to identify and treat this condition at an early stage could result in fatal consequences. Imaging plays an important role in the diagnosis and management of pseudocyst occurring after splenectomy. The advent of interventional radiology has provided better treatment option for patients with solitary pancreatic pseudocysts with success rates similar to those with open surgery but with lower morbidity and mortality rates. However, its role in the management of multiple pseudocysts remains to be defined. PMID- 11405567 TI - Radiological features of unusual ingested foreign bodies. AB - For patients who ingest foreign bodies, pre-operative radiography is an invaluable tool that allows the surgeon to confirm the presence of the foreign body, preempt potential difficulties and plan the best method for foreign body removal. This is particularly important when a foreign body is of usual shape and size. We present a series of radiographs and pictures of some of the retrieved foreign bodies of five patients who swallowed unusual foreign bodies. PMID- 11405568 TI - The design and assessment of questionnaires in clinical research. AB - Questionnaires are one of the most commonly used tools for data collection in clinical research. Despite its simplicity and convenience of use, the design of questionnaire instruments that accurately measure health status and their determinants is nevertheless a difficult and challenging task.We review the two most important issues which are reliability and validity. Reliability can be defined as the degree to which a measure gives 'consistent' or 'reproducible' values when applied in different situations. Validity refers to the extent in which the true value of a variable is correctly measured by the instrument. For different types of questionnaire measurement instruments, specific issues of content, construct and criterion validity should be appropriately addressed. Accuracy in questionnaire-based measurement in clinical studies is achieved by paying attention to the relevant specific issues of reliability and validity during development and testing of such questionnaires. PMID- 11405569 TI - Internet resources for practice and teaching of evidence based medicine. PMID- 11405570 TI - Clinics in diagnostic imaging (59). Crossed-fused renal ectopia. AB - A 62-year-old man was incidentally found to have a palpable right paraumbillcal mass. Ultrasonography and intravenous urogram showed cross-fused renal ectopia. The embryology and clinical features of renal ectopia are discussed. PMID- 11405571 TI - Meta-analysis of theory-of-mind development: the truth about false belief. AB - Research on theory of mind increasingly encompasses apparently contradictory findings. In particular, in initial studies, older preschoolers consistently passed false-belief tasks-a so-called "definitive" test of mental-state understanding-whereas younger children systematically erred. More recent studies, however, have found evidence of false-belief understanding in 3-year-olds or have demonstrated conditions that improve children's performance. A meta-analysis was conducted (N = 178 separate studies) to address the empirical inconsistencies and theoretical controversies. When organized into a systematic set of factors that vary across studies, false-belief results cluster systematically with the exception of only a few outliers. A combined model that included age, country of origin, and four task factors (e.g., whether the task objects were transformed in order to deceive the protagonist or not) yielded a multiple R of .74 and an R2 of .55; thus, the model accounts for 55% of the variance in false-belief performance. Moreover, false-belief performance showed a consistent developmental pattern, even across various countries and various task manipulations: preschoolers went from below-chance performance to above-chance performance. The findings are inconsistent with early competence proposals that claim that developmental changes are due to tasks artifacts, and thus disappear in simpler, revised false-belief tasks; and are, instead, consistent with theoretical accounts that propose that understanding of belief, and, relatedly, understanding of mind, exhibit genuine conceptual change in the preschool years. PMID- 11405572 TI - The future of theory-of-mind research: understanding motivational states, the role of language, and real-world consequences. AB - Suggestions are made for future investigation of theory-of-mind development. There needs to be (1) more focus on the development of understanding of desire and intention; (2) research on the role of language in theory-of-mind development that integrates representational-development and social-interaction views; and (3) investigation of the real-world consequences of children's developing understanding of belief, desire, and intention. PMID- 11405573 TI - Executive accounts of theory-of-mind development. AB - Two varieties of executive theories may be distinguished: emergence accounts and expression accounts. The meta-analytic findings are fully consistent with emergence accounts of theory of mind and do not entirely rule out expression accounts. PMID- 11405574 TI - Reasoning about beliefs: a human specialization? AB - A recent meta-analysis performed by Wellman, Cross, and Watson clears the air surrounding young children's performance on tests of false belief by showing that it is highly likely that there is some type of conceptual development between 3 and 5 years of age that supports improved task performance. The data concerning the evolutionary origin of these abilities, however, is considerably less clear. Nonetheless, there is some reason to suspect that theory of mind is unique to our species, and that its original function was to provide a more abstract level of describing ancient behavioral patterns (such as deception, reconciliation, and gaze following)-behaviors that humans share in common with many other species. Thus, the initial selective advantage of theory of mind may have been because it increased the flexibility of already-existing behaviors, not because it generated scores of radically new ones. PMID- 11405575 TI - Minds, modules, and meta-analysis. AB - Wellman and colleagues' meta-analysis of performance on the false-belief task is methodologically useful, but it does not lead to any theoretical progress concerning the nature of the mechanisms that underlie the existence and development of "theory of mind." In particular, the results of this meta-analysis are perfectly compatible with "early competence" accounts that posit a specific, innate, and possibly modular basis for theory of mind. The arguments presented by Wellman and colleagues against such views stem not from their meta-analytic data, but from mistaken assumptions regarding the requirements of such theories (e.g., that there exist manipulations that improve performance only, or to a greater degree, in young children). Contrary to what Wellman and colleagues claim, their meta-analysis, while consistent with conceptual change, does not lend any new support for such theories. PMID- 11405576 TI - Theory of mind and conceptual change. AB - We agree with the commentaries by Scholl and Leslie, and also by Moses, that the meta-analytic findings do not definitively rule out early competence accounts. But they do make extant versions of such accounts increasingly unlikely. In particular, the meta-analytic findings argue against executive function expression accounts, including the Theory-of-Mind Mechanism/Selection Processor account advocated by Scholl and Leslie. Specifically, Scholl and Leslie articulate two explicit predictions of their account: that task manipulations that attenuate inhibitory demands should differentially advantage older children, and that theory-of-mind developments should occur with consistent timetables. Both of these specific predictions are clearly contradicted, not supported, by the meta-analytic findings. PMID- 11405577 TI - Infants parse dynamic action. AB - As observers of human behavior, infants are faced with a complex flow of motion in which pauses are rare and only occasionally coincide with boundaries between intentional actions. Two studies investigated whether, despite such complexity, 10- to 11-month-old infants (N = 16 for each study) possess skills for parsing ongoing behavior along boundaries correlated with the initiation and completion of intentions. After being familiarized with digitized sequences of continuous everyday action, infants showed renewed interest in test versions in which motion paused in the midst of an actor's pursuit of intentions (interrupting test videos). In contrast, pauses that suspended motion at intention boundary points (completing test videos) sparked no such renewed interest on infants' part. Moreover, basic salience differences between the two types of test videos were not the source of infants' increased interest when intentions were interrupted (Study 2). These findings demonstrate that infants readily detect disruptions of the structure inherent in intentional action, and hence parse ongoing behavior with respect to such structure. Such parsing skill is likely a prerequisite to the development of genuine intentional understanding. PMID- 11405578 TI - Evidence for referential understanding in the emotions domain at twelve and eighteen months. AB - Infants as young as 12 months readily modulate their behavior toward novel, ambiguous objects based on emotional responses that others display. Such social referencing skill offers powerful benefits to infants' knowledge acquisition, but the magnitude of these benefits depends on whether they appreciate the referential quality of others' emotional messages, and are skilled at using cues to reference (e.g., gaze direction, body posture) to guide their interpretation of such messages. Two studies demonstrated referential understanding in 12- and 18-month-olds' responses to another's emotional outburst. Infants relied on the presence versus absence of referential cues to determine whether an emotional message should be linked with a salient, novel object in the first study (N = 48), and they actively consulted referential cues to disambiguate the intended target of an affective display in the second study (N = 32). These findings provide the first experimental evidence of such sophisticated referential abilities in 12-month-olds, as well as the first evidence that infant social referencing at any age actually trades on referential understanding. PMID- 11405579 TI - A further look at the prognostic power of young children's reports of depressed mood and feelings. AB - A primary objective of the present study was to determine the validity of first graders' self-reports of depressed mood and feelings. To that end, the prognostic power of first grade self-reports of depressed mood and feelings was examined with respect to later psychopathology and adaptive functioning in a population of urban school children (N = 946). First grade self-reports of depressed mood predicted later child academic functioning, the need for and use of mental health services, suicidal ideation, and a diagnosis of major depressive disorder by age 14. The prognostic power of these early self-reports suggests that children as young as 5 or 6 years of age are capable of providing valid reports of depressed mood and feelings. PMID- 11405580 TI - Maternal responsiveness and children's achievement of language milestones. AB - This prospective longitudinal study examined the contribution of dimensions of maternal responsiveness (descriptions, play, imitations) to the timing of five milestones in children's (N = 40) early expressive language: first imitations, first words, 50 words in expressive language, combinatorial speech, and the use of language to talk about the past. Events-History Analysis, a statistical technique that estimates the extent to which predictors influence the timing of events, was used. At 9 and 13 months, maternal responsiveness and children's activities (e.g., vocalizations, play) were coded from videotaped interactions of mother-child free play; information about children's language acquisition was obtained through biweekly interviews with mothers from 9 through 21 months. Maternal responsiveness at both ages predicted the timing of children's achieving language milestones over and above children's observed behaviors. Responsiveness at 13 months was a stronger predictor of the timing of language milestones than was responsiveness at 9 months, and certain dimensions of responsiveness were more predictive than others. The multidimensional nature of maternal responsiveness and specificity in mother-child language relations are discussed. PMID- 11405581 TI - Learning proper and common names in inferential versus ostensive contexts. AB - A single, indirect exposure to a novel word provides information that could be used to make a fast mapping between the word and its referent, but it is not known how well this initial mapping specifies the function of the new word. The four studies reported here compare preschoolers' (N = 64) fast mapping of new proper and common names following an indirect exposure requiring inference with their learning of new names following ostension. In Study 1, 3-year-olds were shown an animate-inanimate pair of objects and asked to select, for example, Dax, a dax, or one. Children spontaneously selected an animate over an inanimate object as the referent for a novel proper name, but had no animacy preference in common name or baseline conditions. Next, the children were asked to perform actions on, for example, Dax or a dax, when presented with an array of three objects: the one they had just selected, another member of like kind, and a distracter. An indirectly learned proper name was treated as a marker for the originally selected object only, whereas a new common name was generalized to include the other category member. Study 2 showed that mappings made by inference were as robust as those made by ostension. Studies 3 and 4 demonstrated that even 2-year-olds can learn as much about the function of a new word from an indirect exposure as from ostension. PMID- 11405582 TI - Learning proper nouns and common nouns without clues from syntax. AB - Syntax has been noted to play an important role in word learning in English; it distinguishes the fundamental conceptual difference between individuals (coded as proper nouns), nonindividuals (coded as mass nouns), and classes of individuals (coded as count nouns). The Japanese language does not have grammatical markers flagging the distinctions between count nouns and mass nouns, between proper nouns and common nouns, or between singulars and plurals. How Japanese 2- and 4 year-olds assign meaning to novel nouns associated with familiar and unfamiliar animals and inanimate objects was studied in the research reported here. When a novel label was given to an unfamiliar object, children assumed it to be a name for a basic-level object category whether the referent was an animal or an inanimate object. If the named object already had an established name, and if the object was an inanimate object, the children mapped the noun to a subordinate category. When the named object was an animal, however, they tended to interpret the label as a proper name. These results demonstrated that in the absence of useful information from syntax, 2-year-old Japanese children are able to fast map a noun to its meaning by elegantly coordinating word-learning biases and other available sources of information. PMID- 11405583 TI - Preschool children's difficulty understanding the types of information obtained through the five senses. AB - Two studies explored 3- and 4-year-olds' (N = 60) understanding that the five senses can each lead to different types of knowledge. In Study 1, 40 children engaged in five scenarios in which they could only perform one sensory action to identify the property of an object (e.g., color, scent). After performing the action, children were asked how they found out the property and to show the experimenter how they had found it out. Using a Mr. Potato Head doll, children were also asked to indicate the sensory organ the doll would need to use to identify the property. In Study 2, 20 children presented with five Mr. Potato Head dolls, each sporting only one sensory organ (e.g., a nose), were asked which Mr. Potato Head could find out the property in question. The 3-year-olds performed significantly poorer than the 4-year-olds on all tasks, suggesting a marked transition in children's ability to recognize the origin of their modality specific knowledge during the time period between 3 and 4 years of age. PMID- 11405584 TI - Early reading development in children at family risk for dyslexia. AB - In a 3-year longitudinal study, middle- to upper-middle-class preschool children at high family risk (HR group, N = 67) and low family risk (LR group, N = 57) for dyslexia (or reading disability, RD), were evaluated yearly from before kindergarten to the end of second grade. Both phonological processing and literacy skills were tested at each of four time points. Consistent with the well known familiarity of RD, 34% of the HR group compared with 6% of the LR group became RD. Participants who became RD showed deficits in both implicit and explicit phonological processing skills at all four time points, clearly indicating a broader phonological deficit than is often found at older ages. The predictors of literacy skill did not vary by risk group. Both risk groups underwent a similar developmental shift from letter-name knowledge to phoneme awareness as the main predictor of later literacy skill. This shift, however, occurred 2 years later in the HR group. Familial risk was continuous rather than discrete because HR children who did not become RD performed worse than LR non-RD children on some phonological and literacy measures. Finally, later RD could be predicted with moderate accuracy at age 5 years, with the strongest predictor being letter-name knowledge. PMID- 11405585 TI - Children's understanding of emotion in speech. AB - Children's understanding of emotion in speech was explored in three experiments. In Experiment 1, 4- to 10-year-old children and adults (N = 165) judged the happiness or sadness of the speaker from cues conveyed by propositional content and affective paralanguage. When the cues conflicted (i.e., a happy situation was described with sad paralanguage), children relied primarily on content, in contrast to adults, who relied on paralanguage. There were gradual developmental changes from 4-year-olds' almost exclusive focus on content to adults' exclusive focus on paralanguage. Children of all ages exhibited greater response latencies to utterances with conflicting cues than to those with nonconflicting cues, indicating that they processed both sources of emotional information. Children accurately labeled the affective paralanguage when the propositional cues to emotion were obscured by a foreign language (Experiment 2, N = 20) or by low-pass filtering (Experiment 3, N = 60). The findings are consistent with children's limited understanding of the communicative functions of affective paralanguage. PMID- 11405586 TI - Analytic and heuristic processing influences on adolescent reasoning and decision making. AB - The normative/descriptive gap is the discrepancy between actual reasoning and traditional standards for reasoning. The relationship between age and the normative/descriptive gap was examined by presenting adolescents with a battery of reasoning and decision-making tasks. Middle adolescents (N = 76) performed closer to normative ideals than early adolescents (N = 66), although the normative/descriptive gap was large for both groups. Correlational analyses revealed that (1) normative responses correlated positively with each other, (2) nonnormative responses were positively interrelated, and (3) normative and nonnormative responses were largely independent. Factor analyses suggested that performance was based on two processing systems. The "analytic" system operates on "decontextualized" task representations and underlies conscious, computational reasoning. The "heuristic" system operates on "contextualized," content-laden representations and produces "cognitively cheap" responses that sometimes conflict with traditional norms. Analytic processing was more clearly linked to age and to intelligence than heuristic processing. Implications for cognitive development, the competence/performance issue, and rationality are discussed. PMID- 11405587 TI - Dyadic analyses of friendship in a sample of preschool-age children attending Head Start: correspondence between measures and implications for social competence. AB - Friendships among a large sample of preschool-age children (N = 471) attending Head Start were assessed. Based on sociometric data, friendship dyads were identified as reciprocated (mutual choice) or nonreciprocated (unilateral choice). Dyads were further classified with respect to gender composition as either same- or mixed-gender dyads. Older children were more likely to participate in a reciprocated friendship than were younger children and reciprocated dyads were more likely to be same-gender than were nonreciprocated dyads. Analyses of interaction between dyad partners revealed that reciprocated friends interacted more frequently across all categories of interaction coded and looked at each other more frequently than did members of nonreciprocated dyads. For the positive interaction subscore, the friendship status effect was modified by a significant interaction with gender composition such that significant effects of friendship status were obtained only for same-gender dyads. Additional analyses indicated that the average social competence level was greater for reciprocated dyads than for nonreciprocated dyads. The findings suggest that reciprocated friendships are meaningful for preschool-age children and may serve as special socialization contexts in which the repertoire of behavior can be exercised and perhaps improved. They also highlight the salience of same-gender friendships in the preschool classroom. PMID- 11405588 TI - Evidence that children and adolescents have internal models of peer interactions that are gender differentiated. AB - This study examined whether children's internal representations reflect gender differences that have been found in peer interactions. The dimensions examined were (1) preferences for dyadic or group situations, (2) whether children who are friends with a given target child are likely to be friends with each other, and (3) perceptions of the probability of knowing information about friends. Participants from preschool; grades 2, 6, 8, and 10; and college (N = 278) were asked questions about typical girls and boys. Results indicate that both girls and boys (1) rate typical boys as preferring group interactions more than do typical girls, a difference present as early as preschool; (2) rate typical boys as more likely than typical girls to be friends with one another if they are friends with the same target boy or girl respectively; and (3) rate typical girls as more likely than typical boys to know certain types of information about friends. These results are consistent with the existence of internal models of social interactions that are at least partially gender specific. PMID- 11405589 TI - A meta-analysis of meausures of self-esteem for young children: a framework for future measures. AB - The objective of this study was to synthesize information from literature on measures of the self in young children to create an empirical framework for developing future methods for measuring this construct. For this meta-analysis, all available preschool and early elementary school self-esteem studies were reviewed. Reliability was used as the criterion variable and the predictor variables represented different aspects of methodology that are used in testing an instrument: study characteristics, method characteristics, subject characteristics, measure characteristics, and measure design characteristics. Using information from two analyses, the results indicate that the reliability of self-esteem measures for young children can be predicted by the setting of the study, number of items in the scale, the age of the children being studied, the method of data collection (questionnaires or pictures), and the socioeconomic status of the children. Age and number of items were found to be critical features in the development of reliable measures for young children. Future studies need to focus on the issues of age and developmental limitations on the complicated problem of how young children actually think about the self and what methods and techniques can aid in gathering this information more accurately. PMID- 11405590 TI - Parental coping with children's negative emotions: relations with children's emotional and social responding. AB - This study examined the relation between parents' reactions to children's negative emotions and social competence. Additionally, the role of parental emotional distress in children's emotional socialization was examined. The emotional reactions of 57 preschoolers (33 girls, 24 boys; M age = 59.2 months) were observed during their free-play interactions. Parents (mostly mothers) completed questionnaires about their reactions to children's negative emotions. An index of children's social competence was obtained from teachers. Results indicated that the relation between harsh parental coping strategies and children's emotional responding was moderated by parental distress. In addition, the relation of the interaction of parental coping and distress to children's social competence was mediated by children's level of emotional intensity. It was concluded that distressed parents who use harsh coping strategies in response to children's negative emotions have children who express emotion in relatively intense ways. In turn, these children find it relatively difficult to behave in a socially competent manner. PMID- 11405591 TI - The influence of group size on children's competitive behavior. AB - The present research was designed to test the hypothesis that children would compete more in tetrads than in dyads. Twenty-two pairs of male and 14 pairs of female target children (N = 72) played a competitive game in both tetrads and dyads. Consistent with the hypothesis, male target children competed more in tetrads than in dyads. This hypothesis was not supported for females, however. Analyses of the dynamics of tetrads and dyads further demonstrated that based on a global measure of smiling, the emotional atmosphere was less positive in tetrads than in dyads. The causes and consequences of interaction in different sized social groups are discussed. PMID- 11405592 TI - Negotiating the transition to middle school: the role of self-regulatory processes. AB - The present research examined the role of maladaptive self-regulatory beliefs as vulnerability factors for academic and emotional difficulties during the transition to middle school. A short-term longitudinal design was employed to follow two groups of early adolescents: 187 adolescents who experienced a school transition between the fifth and sixth grades, and 142 adolescents who did not experience a school transition between the fifth and sixth grades. Adolescents completed measures of perceptions of academic control and importance of academic success, experience of chronic academic strain, daily school hassles, and depressive symptoms. Teachers reported on students' academic engagement, including levels of helpless behavior, effort, and academic performance. Consistent with the proposed model of self-regulation, maladaptive self regulatory beliefs (i.e., decreased perceptions of academic control and importance) predicted individual differences in perceived school-related stress and depressive symptoms over the course of the middle school transition, but were not associated with academic and emotional difficulties in adolescents who remained in a stable school environment. Moreover, a self-regulatory sequence was identified proceeding from maladaptive self-regulatory beliefs, to academic disengagement, to enhanced perceptions of school-related stress, to depressive symptoms. This study bridges prior theory and research concerning the psychological impact of normative developmental transitions, the developmental context of depression, and the associations among self-regulatory beliefs, achievement-related behavior, and emotional experience. PMID- 11405593 TI - The medical ethos and social responsibility in clinical medicine. AB - The medical profession will face many challenges in the new millennium. As medicine looks forward to advances in molecular genetics and the prospect of unprecedented understanding of the causes and cures of human disease, clinicians, scientists and bioethicists may benefit from reflection upon the origins of the medical ethos and its relevance to postmodern medicine. Past distortions of the medical ethos, such as Nazism and the Tuskegee Syphilis Study, as well as more recent experience with the ethical challenges of employer-based market driven managed care, provide important lessons as medicine contemplates the future. Racial and ethnic disparities in health status and access to care serve as a reminders that the racial doctrines that fostered the horrors of the Holocaust and the Tuskegee Syphilis Study have not been completely removed from contemporary thinking. Inequalities in health status based on race and ethnicity, as well as socioeconomic status, attest to the inescapable reality of racism in America. When viewed against a background of historical distortions and disregard for the traditional tenets of the medical ethos, persistent racial and ethnic disparities and health and the prospect of genetic engineering raise the specter of discrimination because of genotype, a postmodern version of "racist medicine" or of a "new eugenics." There is a need to balance medicine's devotion to the wellbeing of the patient and the primacy of the patient-physician relationship against with the need to meet the health care needs of society. The challenge facing the medical profession in the new millennium is to establish an equilibrium between the responsibility to assure quality health care for the individual patient while affecting societal changes to achieve "health for all." PMID- 11405594 TI - High-risk sexual behaviors among adolescents engaged through a street-based peer outreach program--(the Adolescent HIV Project). AB - On-the-street peer based programs can overcome barriers and successfully engage teens in HIV counseling and testing. This initiative combines on-the-street peer outreach with on-the-street HIV testing in a mobile counseling and testing van. A survey was conducted to measure HIV risk behaviors concern about HIV infection. In year one, the program engaged 1550 youth. Of these, 666 completed HIV counseling and testing. Only 18% indicated that they had not had unprotected sexual intercourse in the preceding year. Thirty-nine percent of the males and 52% of the females had caused or been pregnant. Sixty-six percent of the males and 53% of the females believed that they could become HIV infected. Fifty-three percent of the male and 75% of the female respondents had had a previous HIV test. However concern about HIV did not significantly decrease the prevalence of HIV risk behaviors. Peer outreach and on-the-street counseling and testing is a successful method of increasing HIV testing among high risk youth. Youth are concerned about HIV infection but that concern does not translated into a change in risk behaviors. PMID- 11405595 TI - Mucoepidermoid carcinoma of the oral cavity. AB - There is presently no uniformly accepted grading system for mucoepidermoid carcinoma, largely due to a lack of consensus as to what criteria should be used to formulate histological grades. The present study was undertaken to determine the relationship between histological grade, clinical stage and survival in these neoplasms. Clinical and histological data from 34 patients with mucoepidermoid carcinoma were reviewed. Mucoepidermoid carcinoma was most common in the parotid gland (44.1%), while 25% of patients had tumors in the minor salivary glands. Low, intermediate, and high-grade neoplasms accounted for 61.7%, 26.5%, and 11.8% of tumors, respectively. There was a general trend towards increasing clinical aggressiveness with increasing histological grade. Similarly, postoperative tumor recurrences were marginally more common in high-grade than in low-grade mucoepidermoid carcinomas. However, both of these findings were statistically insignificant, mainly due to small sample size, late clinical presentation, poor clinical follow-up, incomplete management and incomplete records. These factors explain the relatively low survival figures in the present study, as compared to higher survival figures in white patients with mucoepidermoid carcinoma. PMID- 11405596 TI - Russell-Silver Syndrome in a Nigerian infant with intrauterine growth retardation. AB - Russell-Silver Syndrome (RSS) is a rare cause of pre-natal dwarfism, associated with recognizable dysmorphic features and limb asymmetry. The propositus was a term infant of unrelated Nigerian parents, whose 35-year-old mother had peri conceptual haloperidol for schizophrenia. Anthropometric values suggested severe prenatal stunting in a term infant with asymmetric "head sparing" intrauterine growth retardation (IUGR). A syndromic consideration of Russell-Silver dwarfism was subsequently predicated on the distinctive dysmorphic craniofacial features of a triangular facial profile with a broad forehead and hypoplastic mandible, right upper and lower limb rhizomelia, clinodactyly of the little fingers, micro penis, and (unilateral) cryptochidism. Routine care of a small-for-gestational age infant was pursued, but postnatal growth remained slow (despite adequate caloric provision) until a parent-pressured discharge at 4 weeks. His subsequent demise was said to have occurred "suddenly" 2 weeks post-discharge. Despite the limitations posed by the local paucity of modern investigative tools for genetic disorders, the current case report underscores the diagnostic reality of RSS in a non-white African population. While emphasizing the need for a high index of diagnostic suspicion for congenital malformations and syndromic causes of IUGR in the African sub-region, we suspect a possible etiologic association of haloperidol embryopathy with RSS in the current case. The characteristic features, differential diagnoses, etiologic postulates/current cytogenetic and molecular genetic findings of RSS are fully reviewed in the discussion. PMID- 11405597 TI - Ear pain in children. PMID- 11405598 TI - Detection of anti-ovalbumin IgG in serum by immune complex transfer enzyme immunoassay. AB - An immune complex transfer enzyme immunoassay for anti-ovalbumin IgG in serum is described. Serum-specific antibody was reacted simultaneously with 2,4 dinitrophenyl-bovine serum albumin-ovalbumin conjugate and ovalbumin-peroxidase conjugate. The complex formed by the three components was trapped onto polystyrene balls coated with anti-2,4-dinitrophenyl group IgG, eluted with epsilonN-2,4-dinitrophenyl-L-lysine and transferred to polystyrene balls coated with anti-human IgG-gamma-chain. Bound peroxidase activity was determined by fluorometry. This enzyme immunoassay was 300- to 1,000-fold more sensitive and more reliable than the enzyme-linked immunosorbent assay (ELISA). Anti-ovalbumin IgG was detected in 100% of healthy subjects using this method while only 14% were detected by ELISA. PMID- 11405599 TI - Screening of blood donations by hepatitis C virus polymerase chain reaction (HCV PCR) improves safety of blood products by window period reduction. AB - Introduction of the nucleic acid amplification technique (NAT) as a screening test for blood donors to detect HCV RNA became mandatory on 1 April 1999. Few automated commercial systems are available for HCV RNA detection at the moment. The Cobas Amplicor HCV 2.0 System is able to perform fully automated amplification and detection of nucleic acids. A concentration of 98 IU HCV RNA/ml can be detected by the Cobas Amplicor HCV 2.0 System (n = 233, in 100% of the cases). With a pool size of 40 donor samples, the guidelines of the Paul-Ehrlich Institute concerning sensitivity (5,000 IU HCV RNA per mL in a single donation) were followed. One whole blood donation was identified as HCV-RNA positive (anti HCV IgG negative, GPT < 30 U/L) during a period of 5 months. No false positive test results could be observed. The internal control and the run control are primarily helpful to monitor methodological problems. PMID- 11405600 TI - Simplicity through complexity: immunoblot with recombinant antigens as the new gold standard in Epstein-Barr virus serology. AB - Immunofluorescence tests have been the gold standard in Epstein-Barr virus (EBV) serology during the past decades. Despite their high quality they cannot resolve significant problems with i) anticellular reactivity of some sera, ii) missing anti-EBNA-1 response in 5% of healthy individuals, and iii) loss of anti-EBNA-1 during immunosuppression. Immunoblots with recombinant EBV antigens (either in the Blot or the Line assay version) allow unequivocal serological diagnosis also in those cases that are problematic for immunofluorescence testing, 1) since due to the use of purified recombinant antigens, anticellular reactivities are no problem, and 2) since a combination of two late markers is tested (p72 and p18). Positive p18-IgG thereby defines those cases of past infections in which p72-IgG (anti-EBNA-1) is missing or has been lost. In nearly all cases, a single IgG test is sufficient for a definitive serological diagnosis. Whenever required, the immunoblot technique allows precise quantitation and avidity determination. The immunoblot with recombinant antigens represents the most sensitive, unequivocal and simple test system available at present, with the highest complexity of information obtained. Therefore, the immunoblot with recombinant antigens should be regarded as the new gold standard in EBV serology. PMID- 11405601 TI - Effect of immunotherapy on eosinophil activation in pollen sensitive children. AB - Immunotherapy is a well documented method for treatment of children with allergic airway diseases and allergic rhinitis. The aim of this investigation was to establish the effect of immunotherapy on the presence and activation state of eosinophils in pollen sensitive patients, measured by eosinophil counts and their degranulation product in serum, eosinophil cationic protein in comparison with the (specific) IgE concentration in pollen sensitive patients. In both groups intra-individual variations in (specific) IgE concentrations, due to increase of pollen exposure, were not observed. Blood eosinophil counts and serum ECP concentrations showed consistent results after starting immunotherapy. Overall release of ECP per eosinophil remained also unaffected after the start of immunotherapy. Results of ECP concentrations and eosinophil counts in the subjects' group did not show any deviation when compared with the control subjects' group. It is concluded that the laboratory parameters are not helpful in monitoring the efficacy of immunotherapy in patients with seasonal rhinitis. PMID- 11405602 TI - Review of the epidemiology, molecular characterization and tropism of the hepatitis G virus/GBV-C. AB - The hepatitis G virus and GBV-C are recently discovered variants of the same virus belonging to the family Flaviviridae (HGV/GBV-C). Although initially thought to be a hepatitis virus, it has been shown to have no association with liver disease. This paper reviews the data relating to the discovery, global prevalence, natural history, disease association, molecular features, replication and tissue tropism of HGV/GBV-C. PMID- 11405603 TI - Comparison of monoclonal and polyclonal anti-P1 reagents. AB - Whereas monoclonal reagents for ABO, Rhesus, Kell, Kidd, Lewis and MNS antigens are widely used in blood grouping laboratories and have been well evaluated, little attention has been given so far to monoclonal anti-P1 reagents. Therefore it was the aim of our study to examine the suitability of monoclonal anti-P1 reagents for routine use in comparison to polyclonal anti-P1 reagents. 10 polyclonal anti-P1 reagents (9 from goat, 1 from rabbit) and 5 monoclonal anti-P1 reagents (4 with clone 650, 1 with clone OSK17) were tested by the tube-spin method at room temperature (incubation time as prescribed by the manufacturer) using untreated P1+ red blood cells (RBCs) for titration and untreated P1- RBCs as negative controls. P1- RBCs with a positive direct anti-globulin test (DAT+) (coated with incomplete anti-D) and sialidase-treated P1- RBCs (T+) were used to recognize false positive reactions. 3 monoclonal anti-P1 reagents had a titer of < or = 1 with P1+(weak) RBCs, but all polyclonal anti-P1 reagents showed a titer > or = 2. Negative controls (untreated P1- RBCs) were inconspicuous. 7 polyclonal and 1 monoclonal anti-P1 reagents showed false positive reactions with P1- DAT+ RBCs. All polyclonal and 1 monoclonal (containing clone OSK17) reagent showed false positive reactions with P1- T+ RBCs. As several monoclonal anti-P1 reagents are less suitable for routine use than polyclonal anti-P1 reagents, quality control of anti-P1 reagents should be intensified. PMID- 11405604 TI - Present-day investigations cannot adequately determine the risk of acute coronary syndromes. AB - It is an important irony that present-day clinical stress testing methods including exercise electrocardiogram, stress echocardiography and even coronary angiography are not able to demonstrate vulnerable coronary plaques at risk of rupture. A vulnerable plaque may in fact be invisible on clinical stress test and perhaps only visualized directly through less available techniques such as coronary angioscopy. Landmark pathological studies have deepened our understanding of the mechanisms behind acute coronary syndromes over the last decade. Thrombosis plays a key role and is a unifying feature in the pathogenesis. Platelet-rich thrombus superimposed over the disrupted atherosclerotic plaque or eroded plaque endothelium, with or without fibrin thrombus extension, is evident in post-mortem necropsy and angioscopic studies. However features which contribute to the risk of acute events lie in the atherosclerotic plaque itself. Plaque content and not plaque size is the important factor. Clinical stress testing demonstrates plaque size but not plaque content. A plaque will be prone to rupture if it has only a thin cap and a proportionally larger lipid core. In such a plaque there is preponderance of activated macrophages and T-lymphocytes, and high activity of matrix metalloproteinases. Smooth muscle cell proliferation and collagen synthesis are downregulated. These features may serve as possible targets for devising clinical methods to detect plaques at risk or for reversing the risk in vulnerable plaques. PMID- 11405605 TI - Cardiac natriuretic peptides: new laboratory parameters in heart failure patients. AB - Natriuretic peptides, atrial natriuretic peptide and brain natriuretic peptide, are key regulators in the homeostasis of salt and water excretion and in the maintenance of blood pressure. During heart failure, these peptides are highly activated because of volume overload and increased myocardial wall tension. Among all natriuretic peptides and neurohormones, brain natriuretic peptide and its N terminal prohormone fragment have been shown to be the best markers to identify patients with heart failure. They are useful prognostic markers as well. The stability of brain natriuretic peptides and N-terminal prohormones in ethylene diamine-tetraacetic-acid whole blood is sufficient for routine use. Sensitive and specific assays without the need for plasma extraction are commercially available. The available data indicate that natriuretic peptides are powerful diagnostic and prognostic markers in heart failure patients. First data on treatment guidance are promising. PMID- 11405606 TI - The determination of free and protein-bound haemoglobin in plasma using a combination of HPLC and absorption spectrometry. AB - The aim of the study was to develop a method for the determination of haemoglobin in plasma suitable for use to set target values for external quality assessment schemes for this analyte using commercially available test kits and equipment. In the early phase of the method development it became clear that the use of a single method, namely HPLC, would not be possible. However, by combining HPLC and absorption spectrophotometry, both qualitative and quantitative rapid determinations of protein-bound and free haemoglobin were able to be performed on equipment present in most routine clinical chemistry laboratories. The separation of protein-bound and free haemoglobin could be carried out using commercial HPLC equipment for the determination of haemoglobin A1c (HbA1c) without modification of the conditions used. Instead of haemolysed blood, the same volume of plasma (10 microl) was injected. The eluate was not discarded, but collected in 1-minute fractions so that the void volume (protein-bound Hb) and the haemoglobin peaks (free Hb) were available for the colorimetric determination of haemoglobin using the pseudoperoxidase activity of the haem moiety on hydrogen peroxide and a chromogen (3,3',5,5'-tetramethylbenzidine) in concentrated acetic acid and optimal determination at 600 nm. (In this publication at 578 nm due to the use of a spectrophotometer with Hg-discharge lamp and filter). The appearance of a blue colour in the reaction tube or cuvette indicated the presence of haemoglobin. The use of the above chromogen, with its absorption maximum around 600 nm excluded interference from serum components such as bilirubin, which may interfere in the conventional method often used to determine plasma haemoglobin. The method can be used quantitatively by including an aqueous human haemoglobin standard in the run. This elutes from the HPLC column only as free haemoglobin in the concentration range from 0.1 to 10 g/l. Addition of human haemoglobin to haemoglobin-free plasma resulted in the binding of all Hb to plasma proteins up to a concentration between 2 and 3 g/l (void-volume fraction). At higher concentrations free Hb appeared in the 3-5 minute fractions. These observations agree with published data on the scavenging capacity of plasma for Hb released from erythrocytes. The method is rapid, (HPLC-run maximally 6 min, quantitative colorimetric results 5-10 min) precise (inter-assay coefficients of variation < 8%) and suitable for answering the question as to whether the protein-binding (scavenging) system which prevents the nephro- and cerebrotoxic effects of haemoglobin has been saturated or not, an important question in patients with acute haemolysis problems. A qualitative result is obtainable within 10 minutes of injecting the sample into the HPLC-system. The use of this assay in controlling blood transfusion and haemolytic events arising from surgery, intravascular haemolytic bacteria or artificial heart valves can help in rapid corrective action, if needed. PMID- 11405607 TI - DNA-binding activity of anti-DNA antibody is inhibited by an inorganic polyanion, undecatungstophosphate. PMID- 11405608 TI - When should digoxin be used in patients with diastolic dysfunction? PMID- 11405609 TI - The promise and reality of nitric oxide in the diagnosis and treatment of lung disease. AB - Endogenously produced nitric oxide plays a major role in lung physiology and pathology. Inhaled nitric oxide given exogenously has been studied extensively as a treatment for many lung diseases, and the results suggest that it may help improve oxygenation in some patients. Several issues need to be addressed, however, before it can be used in routine clinical practice. PMID- 11405610 TI - Is glucosamine an effective treatment for osteoarthritic pain? PMID- 11405611 TI - Microbial drug resistance and the roles of the new antibiotics. AB - Physicians should be cautious in prescribing broad-spectrum antibiotics, particularly vancomycin and the fluoroquinolones, because widespread use of these drugs is promoting antibiotic resistance. Resistance is now found in many organisms, including staphylococci, enterococci, streptococci, pneumococci, and Pseudomonas aeruginosa. Some resistant strains can be treated with alternative narrower-spectrum antibiotics. In addition, five newly licensed antibiotics are available, but they should be used judiciously because of their side effects, high cost, and ability to promote additional resistance. PMID- 11405612 TI - Inflammatory myopathies: narrowing the differential diagnosis. AB - Muscle weakness is a feature of numerous conditions, but the muscle weakness of inflammatory myopathies, especially polymyositis and dermatomyositis, is easy to differentiate by specific clinical, laboratory, electromyographic, and histological features. PMID- 11405613 TI - C-reactive protein: a 'golden marker' for inflammation and coronary artery disease. AB - Numerous studies have shown that elevated levels of C-reactive protein (CRP) are associated with increased cardiovascular risk. We advocate greater use of CRP measurements in clinical practice to identify patients at high risk in a variety of situations. PMID- 11405614 TI - C-reactive protein elevation can be caused by conditions other than inflammation and may reflect biologic aging. AB - Epidemiologic studies have revealed that minimal acute-phase changes predict poor prognoses in many conditions and predict disability and mortality in the elderly. These findings have usually been interpreted to indicate that inflammatory processes of some kind play a role in these situations. In fact, a minimal acute phase response does not necessarily establish the existence of an inflammatory process but may also reflect a variety of noninflammatory states, including obesity, sleep disturbance, depression, chronic fatigue, and low levels of physical activity. I propose that a minimal acute-phase response may also be a marker of biologic aging, a condition known to predispose to poor prognoses and to death. PMID- 11405615 TI - Evidence of smoke and atherosclerotic fire. PMID- 11405616 TI - It's a rave new world: rave culture and illicit drug use in the young. AB - Illicit drug use by young people has changed in the last decade, with the increasing use of "designer" or "club" drugs such as ecstasy. Keeping abreast of current trends in illicit drug use prepares the primary care clinician to recognize the clinical effects of drug use, to manage drug emergencies, and to detect addictive behavior. Today's widely used drugs, their street names, their effects, and how to manage overdoses are reviewed. PMID- 11405617 TI - Patient information. Date rape drugs: what parents should know. PMID- 11405618 TI - West Nile fever: here to stay and spreading. AB - The ultimate extent of West Nile virus's range in North America is uncertain but is likely to expand in 2001. Spread chiefly by night-biting Culex mosquitoes, the virus results in infection that most often is asymptomatic or causes a self limited febrile illness. The elderly, however, are prone to develop neurologic manifestations, including potentially fatal encephalitis. PMID- 11405619 TI - Anticoagulation therapy in an elderly woman with atrial fibrillation. PMID- 11405620 TI - Using aspirin and ACE inhibitors in combination: why the hullabaloo? AB - Observational studies indicate that aspirin may counteract the beneficial effect of angiotensin-converting enzyme (ACE) inhibitors, but the data are not yet sufficient for making firm recommendations. We review the available data and offer tentative conclusions. PMID- 11405621 TI - Analysis of chromosomal and organellar DNA of somatic hybrids between Triticum aestiuvm and Haynaldia villosa Schur. AB - Intergeneric somatic hybridization between wheat (cv. Jinan 177) protoplasts that have 24-28 chromosomes and Haynaldia villosa protoplasts containing 11-14 chromosomes was carried out by the polyethylene glycol (PEG) method. A high frequency of hybrid calli and plants were obtained from the fusion products, as revealed by cytological and biochemical techniques and by PCR analysis of 5S rDNA spacer sequences. GISH (genomic in situ hybridization) analysis confirmed the presence of chromosomes from both parents in the hybrid clones and the common occurrence of translocations between them. The RFLP analysis of the organellar DNA using mitochondrion- and chloroplast-specific probes revealed that mitochondria from both parents existed in the cells of hybrid calli and their recombination, whereas chloroplasts segregated and recombined randomly. The gross morphology of hybrid plants resembled that of wheat, but the gross morphology of their ovaries and anthers were intermediate between those of the two parents. The relationship between hybrid plant regeneration and the balance of genetic materials in hybrid clones is discussed. PMID- 11405622 TI - Abortive assembly of succinate-ubiquinone reductase (complex II) in a ferrochelatase-deficient mutant of Escherichia coli. AB - Heme molecules play important roles in electron transfer by redox proteins such as cytochromes. In addition, a structural role for heme in protein folding and the assembly of enzymes has been suggested. Previous results obtained using Escherichia coli hemA mutants, which are unable to synthesize 5-aminolevulinic acid, a precursor of porphyrins and hemes, have demonstrated a requirement for heme biosynthesis in the assembly of a functional succinate-ubiquinone reductase (SQR or complex II), which is a component of the aerobic respiratory chain. In the present study, in order to investigate the role of the heme in the assembly of E. coli SQR, we used a hemH (encodes ferrochelatase) mutant that lacks the ability to insert iron into the porphyrin ring. The hemH mutant failed to insert functional SQR into the cytoplasmic membrane, and the catalytic portion of SQR [the flavoprotein subunit (Fp) and the iron-sulfur protein subunit (Ip)] was localized in the cytoplasm of the cell. It is of interest to note that protoporphyrin IX accumulated in the mutant cells and inactivated the cytoplasmic succinate dehydrogenase (SDH) activity associated with the catalytic Fp-Ip complex. In contrast, SQR was assembled into the membrane of a heme-permeable hemH double mutant when hemin was present in the culture. Only a low level of SQR activity was found in the membrane when hemin was replaced by non-iron metalloporphyrins: Mn-, Co-, Ni-, Zn- and Cu-protoporphyrin IX, or protoporphyrin IX These results indicate that heme iron is indispensable for the functional assembly of SQR in the cytoplasmic membrane of E. coli, and provide a new insight into the biological role of heme in the molecular assembly of the multi-subunit enzyme complex. PMID- 11405623 TI - An insulator element from the sea urchin Hemicentrotus pulcherrimus suppresses variation in transgene expression in cultured tobacco cells. AB - Specialized DNA sequences known as insulators protect genes from both the positive and negative influences of nearby chromatin. Many insulators have been identified in various species; however, few function in multiple species. We have shown that an insulator from the Ars (arylsulfatase) gene of the sea urchin Hemicentrotus pulcherrimus functions in plant cells. Normally, expression of an introduced chimeric GUS gene is inactivated in approximately 30% of transformed tobacco BY2 clones. Transgenes containing the Ars insulator, however, were expressed in all transformed tobacco BY2 cells. The insulator did not affect the copy number, the chromosomal position of transgene integration or maximum expression levels. These results suggest that the insulator functions to suppress the variation normally associated with transgene expression in tobacco BY2 cells. PMID- 11405624 TI - Partial silencing of the NEC1 gene results in early opening of anthers in Petunia hybrida. AB - The NEC1 gene, previously isolated from Petunia hybrida, is expressed at high levels in nectaries, and in a very localized fashion in stamens, particularly in the anther stomium cells and the upper part of the filament. To elucidate the function of the NEC1 gene, co-suppression was employed for down-regulation of NEC1 expression, and transposon insertion mutagenesis was used to knock out the NEC1 function. Among the transgenic plants and plants carrying dTph1 inserted in the NEC1 gene, an "early open anther" phenotype was observed. In this mutant phenotype, the anthers already open in young flower buds (1.8 cm) that still contain immature pollen, resulting in poor pollen quality and impaired pollen release. The results obtained indicate that NEC1 might be involved in the development of stomium cells, which are ruptured during the normal process of anther dehiscence to release mature pollen. Southern analysis revealed the presence of a highly homologous NEC1-like gene, named NEC2, in the P. hybrida genome. The presence of NEC2 was confirmed by segregation analysis and sequencing of genomic clones. The implications of these results and possible reasons why no visually obvious phenotype in nectaries could be produced by co-suppression or transposon insertion mutagenesis are discussed. PMID- 11405625 TI - A novel Cdc20-related WD-repeat protein, Fzr1, is required for spore formation in Schizosaccharomyces pombe. AB - Ste9/Srw1 which shows sequence homology to Hctl from budding yeast, is an activator of the anaphase-promoting complex (APC) in the fission yeast Schizosaccharomyces pombe. By homology search of the S. pombe genome, we identified the gene fr1+, which encodes the protein with the highest homology to Ste9 among five Cdc20-like proteins. Like Ste9, Fzr1 contains seven WD-repeats in its C-terminal region. In spite of this structural similarity, however, overproduction of either of these proteins cannot complement mutants lacking the other. fzr1+ is transcribed exclusively during meiosis and sporulation, suggesting that it plays a role in these processes. In fact, the fzr1 disruptant formed aberrant asci, which contained only one or two mature spores, though meiotic nuclear divisions proceeded with kinetics similar to wild type, and meiotic segregation of chromosomes was normal. Structural alteration of spindle pole bodies, which is a prerequisite for the formation of the forespore membrane, occurred normally in fzr1delta during the second meiotic division. Localization of spore rim marker proteins fused to green fluorescent protein showed that nascent prespores were irregularly shaped, small in size and few in number in fzr1delta cells compared to wild-type cells. Furthermore, electron microscopy revealed that the outer layer of the spore walls was often missing in fzr1delta spores. These results show that Fzr1 is specifically involved in the assembly of the spore envelope and also in spore maturation. Fzr1, a structural homolog of the APC regulator, therefore plays an important role in spore morphogenesis. PMID- 11405626 TI - dtctex-1, the Drosophila melanogaster homolog of a putative murine t-complex distorter encoding a dynein light chain, is required for production of functional sperm. AB - Tctex-1 is a light chain of the cytoplasmic and flagellar dyneins and a candidate for one of the distorter products that cause transmission ratio distortion in mice. We report the identification, characterization, and a preliminary mutational analysis of the function of the Drosophila melanogaster dtctex-1 gene, the putative ortholog of the mammalian tctex-1 gene family. Four P-transposon insertions which disrupt the 5' untranslated region of dtctex-1 are viable in homozygous form but cause male sterility due to the production of non-motile sperm. In males homozygous for dtctex-1 mutant alleles the dtctex-1 transcript is undetectable, while in homozygous females transcripts of lower molecular weight are present. By secondary mobilization of P-element insertions several revertants and new mutant alleles carrying deletions in the 5' UTR region of the gene were produced and characterized by PCR and by Northern analysis. PMID- 11405627 TI - Organization, expression, and function of Caulobacter crescentus genes needed for assembly and function of the flagellar hook. AB - This paper reports on the organization, expression, and function of the divergently transcribed flbG and flaN operons in the hook gene cluster of Caulobacter crescentus. The transcription initiation site of flbG was determined previously, and in this work the transcription map was completed by locating the 3' end of the mRNA using nuclease S1 protection assays. A previous genetic study had suggested that the flbG operon is comprised of four genes; however, the nucleotide sequence revealed three tandemly arranged ORFs that correspond to 5' flbG, flbH, and flgE. FlbG is similar to FliK proteins which are required for termination of hook synthesis, FlbH is similar to FlgD proteins which are essential scaffolding proteins that cap the hook during its assembly, and FlgE corresponds to the hook structural protein. The divergently transcribed flaN gene codes for a hook associated protein I homolog based on its inferred amino acid sequence similarity to FlgK proteins. Based on the amino acid sequence similarities and phenotypes of mutants, flbG, flbH, and flaN have been renamed fliK, flgD, and flgK, FlgD, FlgE, and FlgK proteins, with apparent molecular masses of 23, 68, and 41 kDa, respectively, were expressed from plasmids in a cell-free coupled transcription-translation system, and a protein corresponding to FliK was identified as part of a 190-kDa FliK-LacZ fusion protein. We present evidence showing that, in addition to its role in termination of hook synthesis, FliK is also required for initiation of hook assembly. PMID- 11405628 TI - Expression of intron-containing GUS constructs is reduced due to activation of a cryptic 5' splice site. AB - An intron-containing beta-glucuronidase (GUS) gene has been used widely in promoter analyses and as a plant transformation marker. Maximal plant gene expression requires accurate and efficient removal of the intron from the expressed pre-mRNA transcripts by splicing. Detailed analysis of splicing of potato ST-LS1 and pea legumin introns from GUS constructs revealed the activation of a cryptic 5' splice site in the GUS coding sequence 4 nt upstream from the authentic intron 5' splice site. About 40% of transcripts utilised the cryptic 5' splice site in tobacco protoplasts, reducing the translational potential of expressed pre-mRNA. The same cryptic splicing event was evident in transgenic tobacco leaves but at reduced levels. Mutations that removed the cryptic 5' splice site are associated with a two-fold enhancement in GUS activity in tobacco protoplasts, highlighting the need for careful examination of introns and their sites of insertion into gene constructs to minimise variability in gene activity and maximise gene expression. PMID- 11405629 TI - Marker structure and recombination substrate environment influence conversion preference of broken and unbroken alleles in Saccharomyces cerevisiae. AB - Double-strand break (DSB)-induced gene conversion was investigated using plasmid x chromosome (P x C) and chromosomal direct-repeat recombination substrates with markers arranged such that functional (selected) products could not arise by longpatch mismatch repair initiated from the DSB. As seen previously with analogous substrates, these substrates yield products with discontinuous conversion tracts, albeit at low frequency. Most conversion tracts were of minimum length, suggesting that heteroduplex DNA (hDNA) is limiting, or that co repair imposes selective pressure against products with more extensive hDNA. When functional products can arise by long-patch mismatch repair, the broken allele is converted in nearly all products. In contrast, in the absence of long-patch mismatch repair, unbroken alleles are frequently converted, and we show that such conversion depends on both marker structure (i.e., long palindromic vs. nonpalindromic insertions) and the chromosomal environment of the recombination substrate. We propose that conversion of unbroken alleles is largely a consequence of the segregation of unrepaired markers, and that differences in mismatch repair efficiency underlie the observed effects of marker structure and chromosome environment on allele conversion preference. PMID- 11405630 TI - Characterization and expression of beta-1,3-glucanase genes in peach. AB - Beta-1,3-glucanase is one of the pathogenesis-related (PR) proteins involved in plant defense responses. A peach beta-1,3-glucanase gene, designated PpGns1, has been isolated and characterized. The deduced amino acid sequence of the product of PpGns indicates that it is a basic isoform (pI 9.8), and contains a putative signal peptide of 38 amino acids but has no C-terminal extension. Amino acid sequence comparisons revealed that PpGns1 is 69% and 67% identical to citrus and soybean beta-1,3-glucanases, respectively. Southern analysis of total genomic DNA also indicates that at least three genes for beta-1,3-glucanases exist in peach, forming a small gene family. Characterization of four additional clones by PCR has identified a second beta-1,3-glucanase gene, PpGns2. PpGns2 has been partially sequenced, and when compared to PpGns1, it shows high sequence homology, 96% and 99% nucleotide identity in the first and (partial) second exons, respectively. The deduced partial sequence of the PpGns2 product displays only two differences from PpGns1 in the signal peptide and one in the (partial) mature protein (141 amino acids). The 5'-flanking promoter regions of these two genes share 90% identity in nucleotide sequences interrupted by five major gaps (4-109 nt long). The promoter region contains various sequences similar to cis regulatory elements present in different stress-induced plant genes. In leaves and stems of peach shoot cultures grown in vitro, PpGns1 is induced within 12 h after exposure to a culture filtrate of Xanthomonas campestris pv. pruni or ethephon. However, it is not induced following treatment with mercuric chloride. PMID- 11405631 TI - A new gypsy-type retrotransposon, RIRE7: preferential insertion into the tandem repeat sequence TrsD in pericentromeric heterochromatin regions of rice chromosomes. AB - A portion of an insertion sequence present in a member of the RIRE3 family of retrotransposons in Oryza sativa L. cv. IR36 was found to have an LTR sequence followed by a PBS sequence complementary to the 3'-end region of tRNAMet, indicative of another rice retrotransposon (named RIRE7). Cloning and sequencing of PCR-amplified fragments that made up all parts of the RIRE7 sequence showed that RIRE7 is a gypsy-type retrotransposon with partial homology in the pol region to the rice gypsy-type retrotransposons RIRE2 and RIRE3 identified in rice previously. Interestingly, various portions of the RIRE7 sequence were homologous to several DNA segments present in the centromere regions of cereal chromosomes. Further cloning and nucleotide sequencing of fragments flanking RIRE7 copies showed that RIRE7 was inserted into a site within a tandem repeat sequence that has a unit length of 155 bp. The tandem repeat sequence, named TrsD, was homologous to tandem repeat sequences RCS2 and CentC, previously identified in the centromeric regions of rice and maize chromosomes. Fluorescence in situ hybridization (FISH) analysis of the metaphase chromosomes of O. sativa cv. Nipponbare showed that both RIRE7 and TrsD sequences were present in the centromere regions of the chromosomes. The presence of RIRE7 and the TrsD sequences in the centromere regions of several chromosomes was confirmed by the identification of several YAC clones whose chromosomal locations are known. Further FISH analysis of rice pachytene chromosomes showed that the TrsD sequences were located in a pericentromeric heterochromatin region. These findings strongly suggest that RIRE7 and TrsD are components of the pericentromeric heterochromatin of rice chromosomes. PMID- 11405632 TI - The X element, a novel LINE transposable element from Drosophila melanogaster. AB - Whilst analysing the nature of repeated DNA sequences in the transition zone between euchromatin and heterochromatin at the base of the X chromosome of Drosophila melanogaster, we discovered a novel transposable element of the LINE class that we have named the X element. Several apparently complete elements have been cloned and analysed, and one has been sequenced. It is 4740 bp long, with a polyadenylation sequence and a run of A residues at one end. It contains two ORFs: the 5' ORF is related to the retroviral gag gene and encodes a protein with cysteine-rich motifs that are thought to form a "zinc-knuckle" in a nucleic-acid binding protein; the 3' ORF encodes a putative reverse transcriptase that includes the conserved domains found in reverse transcriptases from other LINEs and retroviruses. The DNA sequence and the sequences of the predicted gene products are most similar to other LINEs from D. melanogaster, such as the F, jockey, Doc and BS elements. Southern analysis suggests that there are at least 30 copies in the genome and that some elements are polymorphic between different strains. Analysis of the DNA sequence of the euchromatic arms of the Drosophila genome identified five full-length elements and a similar number of elements that were intact at the 3' end but had variable 5' truncations. Sequences flanking two different insertion sites were used to design PCR primers to assess the occupancy of sites in wild-type flies of different geographical origins. Flies that lacked each of the insertions were found, suggesting that the element is an active transposon. PMID- 11405633 TI - Stability and inheritance of methylation states at PstI sites in Pisum. AB - The nuclear genome of pea is heavily methylated and the stability of this methylation pattern is unknown. In this study we investigated the stability of DNA methylation and its contribution to restriction fragment polymorphism as judged by AFLP fragment differences. To do this the results of a conventional AFLP analysis were compared with those obtained by a related procedure performed on pre-amplified DNA which provided a 5-methylcytosine-free DNA template (SDAFLP). Genetic mapping in a recombinant inbred (RI) population showed that polymorphisms attributable to different methylation states of PstI sites were abundant, and generally appeared to be stably inherited, although occasional failures of the inheritance of methylation states have been found. Assessments of genetic diversity by AFLP and SDAFLP were in general agreement with each other and with the currently accepted phylogeny of Pisum, but within cultivated groups the number of differences appeared to be exaggerated by AFLP. The data suggest that epigenetic differences may have played a role in the domestication of pea. PMID- 11405634 TI - Functional analysis of promoter sequences of cephalosporin C biosynthesis genes from Acremonium chrysogenum: specific DNA-protein interactions and characterization of the transcription factor PACC. AB - An analysis of the pcbC promoter from the cephalosporin C-producing filamentous fungus Acremonium chrysogenum was performed using fungal transformants carrying reporter gene fusions. By investigating sequential deletion derivatives of the pcbC promoter region, a DNA fragment was identified, which is responsible for transcriptional activation of the pcbC gene. Sequence analysis of this fragment revealed a consensus binding site for the fungal transcription factor PACC. Gel retardation experiments with crude extracts of A. chrysogenum confirmed the specific binding of a protein to the PACC binding site. The subsequent cloning of the pacC homolog from A. chrysogenum allowed the identification of an ORF of 621 amino acids encoded by four exons. The polypeptide shows about 35% sequence identity to other fungal PACC proteins. A PACC protein fragment synthesized in E. coli was used for in vitro binding assays, and specific binding of the zinc finger transcription factor to its consensus binding sites in the promoter regions of four cephalosporin C biosynthesis genes could be demonstrated. The bi directional promoters of the genes pcbAB-pcbC and cefEF-cefG contain two PACC binding sites each. The data obtained strongly suggest that, in A. chrysogenum, the zinc-finger transcription factor PACC is involved in the transcriptional regulation of the genes involved in cephalosporin C biosynthesis. PMID- 11405635 TI - Molecular genetic analysis of the candidate gene for MOD, a locus required for self-incompatibility in Brassica rapa. AB - The MIP-MOD (for MOD-locus associated Major Intrinsic Protein) gene encodes an aquaporin-like product, and has been reported to be a candidate for the MOD gene which is required for the self-incompatibility response in Brassica rapa. In an antisense suppression experiment designed to investigate the role of MIP-MOD, we found that levels of MIP-MOD mRNA in the stigmas of fourteen antisense transgenics, as well as in the self-incompatible cultivar Osome (Osm), were much lower than in the stigmas of the self-incompatible S8 homozygous (S8) strain. Therefore, we analyzed the molecular structure of the MIP-MOD gene in three B. rapa strains: S8, Osm, and the self-compatible var. Yellow Sarson (YS). Nucleotide sequence analysis of the MIP-MOD genes isolated from the three strains revealed that all three encode the same amino acid sequence and that YS and Osm contain the same MIP-MOD allele, designated MIP-MOD(YS). Analysis of other self incompatible B. rapa strains that are homozygous for the MIP-MOD(YS) allele indicated that high levels of MIP-MOD transcripts are not essential for the self incompatibility response. Furthermore, a MOD mutant generated by gamma irradiation was found to contain a wild-type MIP-MOD gene that is expressed at normal levels. These data suggest that MIP-MOD is not MOD itself. We suggest that this gene should be renamed MLM (for MIP gene linked to MOD). PMID- 11405636 TI - Sequence comparisons among dispersed members of the Brassica S multigene family in an S9 genome. AB - Self-incompatibility (SI) systems prevent self-pollination and promote outbreeding. In Brassica, the SI genes SLG (for S-locus glycoprotein) and SRK (for S-receptor kinase) are members of the S multigene family, which share the SLG-like domain (S domain), which encodes a putative receptor. We have cloned members of the S multigene family from the S9 haplotype of B. campestris (syn. rapa). In addition, eight distinct genomic regions harboring 10 SLG/SRK-like genes were characterized in the present study. Sequence analysis revealed two novel SRK-like genes, BcRK3 and BcRK6 (for B. campestris receptor kinases 3 and 6, respectively). Other genes that were characterized included SFR2 (for S gene family receptor 2), SLR2 (for S locus related gene 2), and a pseudogene. Based on phylogenetic analysis of the nucleotide sequences of the S domain regions, SLG and SRK appear to be distinct from other members of the S multigene family. Linkage analysis showed that most members of the S multigene family are dispersed in the Brassica genome, and that SLR1 (S locus related gene 1) is not linked to the SLR2 in B. campestris. PMID- 11405637 TI - Families of clustered microsatellites in a conifer genome. AB - Microsatellite clustering may account for genetic maps which do not coalesce into the expected number of linkage groups. Microsatellite organization within the large genome of Pinus taeda (1C = 20,000 Mb) was determined by (1) testing whether repeat motifs were sequestered within the low-copy DNA kinetic component and (2) testing for repeat motif clusters within DNA fragments regardless of copy number. Within the low-copy kinetic component, either (AC)n or (AG)n repeat units were present in 32% of sequences. No repeat motifs were found in the total genome control. Clustered repeat motifs were frequent; the (ATG)n triplet repeat motif was located upstream from a CG-rich trinucleotide microsatellite in 26 out of 44 microsatellite sequences. Fourteen of the clustered (ATG)n sequences could be assembled into four microsatellite sequence families based on similarities in the flanking regions. Consistent with the DNA turnover model, family members shared similar flanking regions but differed in repeat motif composition and length. PMID- 11405638 TI - Genetic analyses of correlated solids, flavor, and health-enhancing traits in onion (Allium cepa L.). AB - Onion possesses organosulfur compounds and carbohydrates that provide unique flavor and health-enhancing characteristics. Significant phenotypic correlations have been reported among soluble solids content (SSC), total dry matter, pungency, and onion-induced in vitro antiplatelet activity. A genetic map and segregating F3M families derived from a cross between two inbred populations were used to identify and estimate the effects of quantitative trait loci (QTLs) controlling these traits at 30 and 90 days postharvest. In vitro antiplatelet activities among different onion populations were consistent across six human blood donors. Most of the populations showed in vitro antiplatelet activities; however, for some donors, one of the parental lines and two F3M families had pro aggregatory effects under our experimental conditions. SSC, dry matter, pungency, and in vitro antiplatelet activity showed significant positive phenotypic and genetic correlations. A chromosome region on linkage group E accounted for a significant amount of the phenotypic variation for all of these traits. The correlations among these traits may be due to linkage or pleiotropy of genes controlling solids content. Our results indicate that it will be difficult to develop onion populations with lower pungency and high in vitro antiplatelet activity; however, the strong genetic and phenotypic correlations between high in vitro antiplatelet activity and high SSC are beneficial for the health functionality of onion. PMID- 11405639 TI - Expression of the S receptor kinase in self-compatible Brassica napus cv. Westar leads to the allele-specific rejection of self-incompatible Brassica napus pollen. AB - Expression of an S receptor kinase (SRK910) transgene in the self-compatible Brassica napus cv. Westar conferred on the transgenic pistil the ability to reject pollen from the self-incompatible Brassica napus W1 line, which carries the S910 allele. In one of the SRK transgenic lines, 1C, virtually no seeds were produced when the transgenic pistils were pollinated with W1 pollen (Mean number of seeds per pod = 1.22). This response was specific to the W1 pollen since pollen from a different self-incompatible Brassica napus line (T2) and self pollinations were fully compatible. Westar plants expressing an S locus glycoprotein transgene (SLG910) did not show any self-incompatibility response towards W1 pollen. Transgenic Westar plants resulting from crosses between the 1C SRK transgenic line and three SLG910 transgenic lines were also tested for rejection of W1 pollen. The additional expression of the SLG910 transgene in the SRK910 transgenic plants did not cause any significant further reduction in seed production (Mean seeds/pod = 1.04) or have any detectable effects on the number of pollen grains that adhered to the pistil. Thus, while the allele-specific SLG gene was previously reported to have an enhancing effect on the self incompatibility response, no evidence for such a role was found in this study. PMID- 11405640 TI - A role for Sds3p, a component of the Rpd3p/Sin3p deacetylase complex, in maintaining cellular integrity in Saccharomyces cerevisiae. AB - The SDS3 gene was identified in a suppressor screen for mutations that enhance position-effect silencing in yeast. Cells that are defective in SDS3 have pleiotropic phenotypes, similar to those seen in the absence of the histone deacetylase components Rpd3p and Sin3p, including meiotic defects and improper regulation of the HO gene. To gain further insight into SDS3 function we undertook an epistasis analysis with other SDS genes. We found that sds3 is synthetically lethal in combination with a deletion of the SWI6 (SDS11) gene, which encodes a cell-cycle regulator. sds3 swi6 double mutants do not display a specific cell-cycle arrest phenotype, but instead die due to cell lysis. Constitutive expression of the G1 cyclin gene CLN2 restores viability to an sds3 swi6 strain, as does overexpression of SKT5/ CHS4, which encodes a regulatory subunit of chitin synthase III, and SSD1, a gene previously implicated in ensuring cell-cycle progression and cellular integrity. Significantly, growth in the presence of 1 M sorbitol or overexpression of PKC1 also partially suppresses the lethal phenotype of the sds3 swi6 strain. This lethality in the absence of SWI6 function most probably reflects an important or essential role for Sds3p in the Rpd3p/Sin3p histone deacetylase complex, since RPD3 and SIN3 mutations are also synthetically lethal in combination with swi6 and these phenotypes are also rescued by elevated dosage of SKT5/CHS4, SSD1, or PCK1. Taken together, these data indicate that the transcription factor Swi6p and the Rpd3p-based deacetylase complex act in parallel pathways to activate genes required for cell wall biosynthesis. PMID- 11405641 TI - Secretase targets for Alzheimer's disease: identification and therapeutic potential. PMID- 11405642 TI - Design and synthesis of 2-methyl-2-[4-(2-[5-methyl-2-aryloxazol-4 yl]ethoxy)phenoxy]propionic acids: a new class of dual PPARalpha/gamma agonists. AB - Propionic acid derivative 8, which was designed and synthesized based on putative pharmacophores of known PPARgamma- and PPARalpha-selective compounds, exhibits potent dual PPARalpha/gamma agonist activity as demonstrated by in vitro binding and dose overlap in the newly introduced EOB mouse model for glucose lowering and lipid/cholesterol homeostasis. PMID- 11405643 TI - Potential new antitumor agents from an innovative combination of demethylcantharidin, a modified traditional Chinese medicine, with a platinum moiety. AB - A combination of demethylcantharidin, a modified component of a traditional Chinese medicine (TCM), with a platinum moiety has produced a series of TCM-based platinum compounds [Pt(C(8)H(8)O(5))(NH(2)R)(2)] 1-5, which demonstrate selective cytotoxicity toward SK-Hep-1 (human liver) cell line, and circumvention of cross resistance. The inclusion of demethylcantharidin rendered the compounds highly active as protein phosphatase (PP2A) inhibitors. The new TCM-Pt compounds may possess a novel dual mechanism of antitumor action: inhibition of PP2A and platination of DNA. PMID- 11405644 TI - 3-(4-aroyl-1H-pyrrol-2-yl)-N-hydroxy-2-propenamides, a new class of synthetic histone deacetylase inhibitors. AB - Novel 3-(4-aroyl-2-pyrrolyl)-N-hydroxy-2-propenamides are disclosed as a new class of histone deacetylase (HDAC) inhibitors. Three-dimensional structure-based drug design and conformational analyses into the histone deacetylase-like protein (HDLP) catalytic core suggested the synthesis and biological evaluation of compounds 7a-h. Experimental pK(i) values are in good agreement with VALIDATE predicted pK(i) values of new derivatives. All compounds 7a-h show HDAC inhibitory activity in the micromolar range, with 7e as the most potent derivative (IC(50) = 1.9 microM). The influence of the 4'-substituent in the aroyl moiety is not significant for the inhibitory activity, as all compounds 7a g show IC(50) values between 1.9 and 3.9 microM. Otherwise, the unsaturated chain linking the pyrrole ring to the hydroxamic acid group is clearly important for the anti-HDAC activity, the saturated analogue 7h being 10-fold less active than the unsaturated counterpart 7a. PMID- 11405645 TI - Transformation of a kappa-opioid receptor antagonist to a kappa-agonist by transfer of a guanidinium group from the 5'- to 6'-position of naltrindole. AB - The importance of the indole scaffold of GNTI 3 in directing its address (5' guanidinium group) to associate with the Glu297 residue of the kappa-opioid receptor was investigated by the synthesis and biological evaluation of its 4'- (4a), 6'- (4b), and 7'- (4c) regioisomers. The finding that only the 5' regioisomer (GNTI) possessed potent kappa-opioid antagonist activity and high affinity at kappa-receptors illustrates the importance of the 5'-position in orienting the guanidinium group to the proper recognition locus (Glu 297) for potent kappa-antagonist activity. The discovery that the 6'-regioisomer of GNTI was a potent kappa-agonist, together with the results of site-directed mutagenesis studies that are consistent with association between the 6' guanidinium group and Glu297, suggest that the transition from an inactive to an active state of the kappa-receptor involves a conformational change of TM6. We propose that association of the 6'-guanidinium group of 4b with Glu297 promotes axial rotational motion of transmembrane helix VI which leads to receptor activation via a conformational change of inner loop 3. PMID- 11405646 TI - Adenosine analogues as selective inhibitors of glyceraldehyde-3-phosphate dehydrogenase of Trypanosomatidae via structure-based drug design. AB - In our continuation of the structure-based design of anti-trypanosomatid drugs, parasite-selective adenosine analogues were identified as low micromolar inhibitors of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). Crystal structures of Trypanosoma brucei, Trypanosoma cruzi, Leishmania mexicana, and human GAPDH's provided details of how the adenosyl moiety of NAD(+) interacts with the proteins, and this facilitated the understanding of the relative affinities of a series of adenosine analogues for the various GAPDH's. From exploration of modifications of the naphthalenemethyl and benzamide substituents of a lead compound, N(6)-(1-naphthalenemethyl)-2'-deoxy-2'-(3 methoxybenzamido)adenosine (6e), N(6)-(substituted-naphthalenemethyl)-2'-deoxy-2' (substituted-benzamido)adenosine analogues were investigated. N(6)-(1 Naphthalenemethyl)-2'-deoxy-2'-(3,5-dimethoxybenzamido)adenosine (6m), N(6)-[1-(3 hydroxynaphthalene)methyl]-2'-deoxy-2'-(3,5-dimethoxybenzamido)adenosine (7m), N(6)-[1-(3-methoxynaphthalene)methyl]-2'-deoxy-2'-(3,5 dimethoxybenzamido)adenosine (9m), N(6)-(2-naphthalenemethyl)-2'-deoxy-2'-(3 methoxybenzamido)adenosine (11e), and N(6)-(2-naphthalenemethyl)-2'-deoxy-2'-(3,5 dimethoxybenzamido)adenosine (11m) demonstrated a 2- to 3-fold improvement over 6e and a 7100- to 25000-fold improvement over the adenosine template. IC(50)'s of these compounds were in the range 2-12 microM for T. brucei, T. cruzi, and L. mexicana GAPDH's, and these compounds did not inhibit mammalian GAPDH when tested at their solubility limit. To explore more thoroughly the structure-activity relationships of this class of compounds, a library of 240 N(6)-(substituted)-2' deoxy-2'-(amido)adenosine analogues was generated using parallel solution-phase synthesis with N(6) and C2' substituents chosen on the basis of computational docking scores. This resulted in the identification of 40 additional compounds that inhibit parasite GAPDH's in the low micromolar range. We also explored adenosine analogues containing 5'-amido substituents and found that 2',5'-dideoxy 2'-(3,5-dimethoxybenzamido)-5'-(diphenylacetamido)adenosine (49) displays an IC(50) of 60-100 microM against the three parasite GAPDH's. PMID- 11405647 TI - Development of potent non-carbohydrate imidazole-based small molecule selectin inhibitors with antiinflammatory activity. AB - A novel series of non-carbohydrate imidazole-based selectin inhibitors has been discovered via high-throughput screening using a P-selectin ELISA-based assay system. The initial lead 1 had an IC(50) of 17 microM in the P-selectin ELISA; this potency was significantly improved via an extensive SAR exploration. One of the current lead compounds (29) has an IC(50) of 300 nM in a P-selectin ELISA; it also has good activity in P- and E-selectin cell adhesion assays and shows efficacy in vivo. These compounds represent a novel series of sLe(X) mimetics with antiinflammatory activity. Their unique profile supports our interest in their further evaluation as drug candidates for the treatment of inflammation. Herein we describe the syntheses, optimization, and SAR of this series of novel potent selectin antagonists. PMID- 11405648 TI - Bicyclic analogues of D-myo-inositol 1,4,5-trisphosphate related to adenophostin A: synthesis and biological activity. AB - The high affinity of adenophostin A for 1D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] receptors may be related to an alteration in the position of its 2'-phosphate group relative to the corresponding 1-phosphate group in Ins(1,4,5)P(3). To investigate this possibility, two bicyclic trisphosphates 9 and 10, designed to explore the effect of relocating the 1-phosphate group of Ins(1,4,5)P(3) using a novel fused-ring system, were synthesized from myo inositol. Biological evaluation of 9 and 10 at the Ins(1,4,5)P(3) receptors of hepatocytes showed that both were recognized by hepatic Ins(1,4,5)P(3) receptors and both stimulated release of Ca(2+) from intracellular stores, but they had lower affinity than Ins(1,4,5)P(3). This finding may be explained by considering the three-dimensional structures of 9 and 10 in light of recent studies on the conformation of adenophostin A. PMID- 11405649 TI - Synthesis, biological evaluation, and pharmacophore generation of new pyridazinone derivatives with affinity toward alpha(1)- and alpha(2) adrenoceptors. AB - A series of new pyridazin-3(2H)-one derivatives (3 and 4) were evaluated for their in vitro affinity toward both alpha(1)- and alpha(2)-adrenoceptors by radioligand receptor binding assays. All target compounds showed good affinities for the alpha(1)-adrenoceptor, with K(i) values in the low nanomolar range. The polymethylene chain constituting the spacer between the furoylpiperazinyl pyridazinone and the arylpiperazine moiety was shown to influence the affinity and selectivity of these compounds. Particularly, a gradual increase in affinity was observed by lengthening the polymethylene chain up to a maximum of seven carbon atoms. In addition, compound 3k, characterized by a very interesting alpha(1)-AR affinity (1.9 nM), was also shown to be a highly selective alpha(1) AR antagonist, the affinity ratio for alpha(2)- and alpha(1)-adrenoceptors being 274. To gain insight into the structural features required for alpha(1) antagonist activity, the pyridazinone derivatives were submitted to a pharmacophore generation procedure using the program Catalyst. The resulting pharmacophore model showed high correlation and predictive power. It also rationalized the relationships between structural properties and biological data of, and external to, the pyridazinone class. PMID- 11405650 TI - Discovery of 4-amino-5-(3-bromophenyl)-7-(6-morpholino-pyridin-3-yl)pyrido[2,3 d]pyrimidine, an orally active, non-nucleoside adenosine kinase inhibitor.. AB - Adenosine (ADO) is an endogenous homeostatic inhibitory neuromodulator that reduces cellular excitability at sites of tissue injury and inflammation. Inhibition of adenosine kinase (AK), the primary metabolic enzyme for ADO, selectively increases ADO concentrations at sites of tissue trauma and enhances the analgesic and antiinflammatory actions of ADO. Optimization of the high throughput screening lead, 4-amino-7-aryl-substituted pteridine (5) (AK IC(50) = 440 nM), led to the identification of compound 21 (4-amino-5-(3-bromophenyl)-7-(6 morpholino-pyridin-3-yl)pyrido [2,3-d]pyrimidine, ABT-702), a novel, potent (AK IC(50) = 1.7 nM) non-nucleoside AK inhibitor with oral activity in animal models of pain and inflammation. PMID- 11405651 TI - Synthesis and activities of pyoverdin-quinolone adducts: a prospective approach to a specific Therapy against Pseudomonas aeruginosa. AB - Pseudomonas aeruginosa is particularly resistant to most all the antibiotics presently available, essentially because of the very low permeability of its outer membrane. To overcome this, we synthesized four siderophore-based antibiotics formed by two quinolones - norfloxacin and benzonaphthyridone - bound to the pyoverdin of P. aeruginosa ATCC 15692 via two types of spacer arms: one stable and the other readily hydrolyzable. From the comparison of their antibacterial properties with those of the two unbound quinolones, we reached the following conclusions: (a) The adducts inhibit Escherichia coli's gyrase showing that the dissociation of the compounds is not necessary for their activity. However, the presence of the pyoverdin moiety on the molecule decreases the inhibition activity compared to the antibiotic alone. (b) They facilitate the uptake of (55)Fe using the specific pyoverdin-mediated iron-transport system of the bacterium. No uptake was observed either with P. aeruginosa ATCC 27853, which produces a structurally different pyoverdin, or with P. aeruginosa K690, which is a mutant of P. aeruginosa ATCC 15692 lacking FpvA, the outer-membrane pyoverdin receptor. (c) MIC determinations have shown that only strains P. aeruginosa ATCC 15692 and the derived outer-membrane receptor-producing but pyoverdin-deficient P. aeruginosa IA1 mutant present higher susceptibility to the pyoverdin-quinolone adducts, whereas P. aeruginosa ATCC 27853 and K690 are much more resistant. (d) Growth inhibition by these adducts confirmed these results and showed that the adducts with the hydrolyzable spacer arm have better activity than those with the stable one and that the labile spacer arm adducts present much higher activity than the quinolones alone. These results show clearly that the penetration of the antibiotic into the cells is favored when this latter is coupled with pyoverdin: Only the strains possessing the appropriate outer-membrane receptor present higher susceptibility to the adduct. In this case the antibiotic uses the pyoverdin-mediated iron-transport system. Furthermore, better efficiency is obtained when the spacer arm is labile and favors the antibiotic release inside the cell, allowing better inhibition of gyrase. PMID- 11405652 TI - Synthesis of novel gamma-aminobutyric acid (GABA) uptake inhibitors. 5.(1) Preparation and structure-activity studies of tricyclic analogues of known GABA uptake inhibitors. AB - On the basis of the SAR of a series of known gamma-aminobutyric acid (GABA) uptake inhibitors, including 4 (SKF 89976), new tricyclic analogues have been prepared. These novel compounds are derivatives of nipecotic acid, guvacine, and homo-beta-proline, substituted at the nitrogen of these amino acids by various lipophilic moieties such as (10,11-dihydro-5H-dibenz[b,f]azepin-5-yl)alkoxyalkyl or (10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5-ylidene)alkoxyalkyl. The in vitro values for inhibition of [(3)H]-GABA uptake in rat synaptosomes was determined for each compound in this new series, and it was found that several of the novel compounds showed a high potency comparable with that of the reference compounds 4, 5 (tiagabine), and 6 (CI-966). Several of the novel compounds were also evaluated for their ability in vivo to inhibit clonic seizures induced by a 15 mg/kg (ip) dose of methyl 6,7-dimethoxy-4-ethyl-beta-carboline-3-carboxylate (DMCM). One compound, (R)-1-(2-(2-(10,11-dihydro-5H-dibenz[b,f]azepin-5 yl)ethoxy)ethyl)-3-piperidinecarboxylic acid (23), was selected for further biological investigations and showed a protective index comparable to or slightly better than that of the recently launched anticonvulsant product 5 ((R)-1-(4,4 bis(3-methyl-2-thienyl)-3-butenyl)-3-piperidinecarboxylic acid). PMID- 11405653 TI - Synthesis, cytotoxicity, and antitumor activity of copper(II) and iron(II) complexes of (4)N-azabicyclo[3.2.2]nonane thiosemicarbazones derived from acyl diazines. AB - A series of thiosemicarbazones (TSCs) (bearing a (4)N-azabicyclo[3.2.2]nonane moiety) derived from 3-acylpyridazines, 4-acetylpyrimidines, and 2 acetylpyrazines (1-8) were synthesized as potential antitumor agents. TSCs 1-8 exhibited potent cytotoxic activity against human acute lymphoblastic leukemia CCRF-CEM cells (IC(50) = 0.05-0.77 microM) and colon adenocarcinoma HT-29 cells (IC(50) = 0.011-2.22 microM). Copper II complexes of TSCs 1-8 showed significant improvement in cytotoxic activity against HT-29 cells (IC(50) = 0.004-1.51 microM) by a factor of 3. However, complexation of ligands 1, 2, 4, and 6 with Fe(II) results in lowering of cytotoxic activity by a factor of approximately 7. In clonogenic assays involving human tumor cells of different tumor origins, compounds 5, 7, 8, and their copper complexes 5Cu(II), 7Cu(II), and 8Cu(II) exhibited remarkable cytotoxic activities with mean IC(50) values of 6, 0.18, 1, 1, 0.37, and 0.37 nM, respectively. In particular, the compounds were highly effective against human colon carcinoma and large and small cell lung carcinoma cells. The TSC derivative 5 was evaluated in vivo in nude mice bearing LXFL 529 human large cell lung carcinoma cells. With respect to antitumor activity, application of 30 mg/kg/d resulted in moderate inhibition (42%) of tumor growth. No effect on tumor growth was observed at a dose of 10 mg/kg/d. However, a dose of 40 or 60 mg/kg/d resulted in 50 and 75% death, respectively, in the treated mice, indicating the high toxicity of these compounds. Using human liver microsomes, compound 5 was found to be rapidly and highly metabolized in vitro. In actual fact, only 2% of the unmetabolized compound could be detected in the incubation medium after 5 min. The IC(50) for cell proliferation (0.006-0.022 microM) elicited by these compounds is much lower than that of the inhibition of [(14)C]cytidine incorporation into DNA (0.18-3.32 microM). These compounds are also noncell cycle specific agents. Interestingly, compounds 5, 5Cu(II), and 8 were found to be potent inducers of apoptosis in Burkitt's lymphoma cells. PMID- 11405654 TI - Combinatorial synthesis of cholesterol ester transfer protein-mRNA ligands and screening by nondenaturating gel-electrophoresis. AB - RNA, as one of the biomolecules with the most structural and functional diversity, is an attractive therapeutic target.(1) Employing combinatorial chemistry methods, small peptide ligands were found, which bind to a short RNA with important biological functions. A 23-nt RNA oligonucleotide from the cholesterol ester transfer protein mRNA was chosen as a molecular target.(2) A 27 nt RNA oligonucleotide from the human immunodeficiency virus type-1 (HIV-1) TAR RNA was used to control the binding specificity.(3) Tetrapeptide libraries, composed of the amino acids Lys, Tyr, Leu, Ile, and Arg, with and without C- and N-terminal lysines, were synthesized by a combination of combinatorial and divergent solid-phase synthesis. Gel-shift affinity screening was used to extract the peptides with the best RNA binding properties. The peptide Lys-Tyr-Lys-Leu Tyr-Lys-Cys-NH(2) (1) showing micromolar affinity to its RNA target was characterized with circular dichroism (CD), ultra violet (UV) measurement, and (1)H NMR spectroscopy. PMID- 11405655 TI - Probing the interaction of dengue virus envelope protein with heparin: assessment of glycosaminoglycan-derived inhibitors. AB - A structure-activity relationship study was carried out to facilitate development of inhibitors of dengue virus infectivity. Previous studies demonstrated that a highly charged heparan sulfate, a heparin-like glycosaminoglycan found on the cell surface, serves as a receptor for dengue virus by binding to its envelope protein. Interventions that disrupt this binding effectively inhibit infectivity. A competitive binding assay was developed to screen polyanionic compounds for activity in preventing binding of dengue virus envelope protein to immobilized heparin; compounds tested included drugs, excipients, and larger glycosaminoglycans and their semisynthetic derivatives. Results of this competitive binding assay were used to select agents for detailed evaluation of interactions by surface plasmon resonance spectroscopy, which afforded binding on rates, off-rates, and dissociation constants. From these data, an understanding of the structural requirements for polyanion binding to dengue virus envelope protein has been established. PMID- 11405656 TI - Amphiphilic anionic analogues of galactosylceramide: synthesis, anti-HIV-1 activity, and gp120 binding. AB - We describe the synthesis together with the results of anti-HIV-1 activity and gp120-monolayer binding experiments of new galactosyl amphiphiles, analogues of galactosylceramide, an alternative receptor used by HIV to infect CD4 negative cells. These compounds consist of single- and double-chain amphiphiles containing one or two galactose residues. To favor their clustering into galactosyl-rich microdomains, their molecular structure contains also an amino group or several hydroxyls or anionic groups, such as carboxylate, sulfate, sulfonate, and phosphate. Among the 12 new galactosylated compounds reported, a specific anti HIV activity, although moderate (IC(50) from 10 to 50 microM), was detected only for three of them, i.e., I-GalSer[CO2Na][C14], II-GalSer[C14][C7SO3Na], and II GalSer[C2SO4Na][C14], which contain an anionic group. The marked increase of surface pressure which was observed upon addition of gp120 into the aqueous subphase underneath the monolayers containing these galactolipids indicated gp120 insertion into the monolayers, suggesting that binding of these three derivatives to HIV-1 gp120 may be responsible for their anti-HIV activity. PMID- 11405657 TI - Novel, potent, and selective phosphodiesterase 5 inhibitors: synthesis and biological activities of a series of 4-aryl-1-isoquinolinone derivatives. AB - A novel class of potent and selective phosphodiesterase 5 (PDE5) inhibitors, 4 aryl-1-isoquinolinone derivatives, which have been designed by the comparison of the structure of cGMP and a previously reported 1-arylnaphthalene lignan, was disclosed. Among these compounds, methyl 2-(4-aminophenyl)-1,2-dihydro-1-oxo-7-(2 pyridinylmethoxy)-4-(3,4,5-trimethoxyphenyl)-3-isoquinoline carboxylate dihydrochloride (36a) exhibited potent PDE5 inhibitory activity (IC(50) = 1.0 nM) with high isozyme selectivities (IC(50) ratio: PDE1/PDE5 = 1300, PDE2/PDE5 > 10 000, PDE3/PDE5 > 10 000, PDE4/PDE5 = 4700, PDE6/PDE5 = 28). Compound 36a also showed the most potent relaxant effect on isolated rabbit corpus cavernosum (EC(30) = 7.9 nM). Compound 63 (T-1032), the sulfate form of 36a, was selected for further biological and pharmacological evaluation of erectile dysfunction. PMID- 11405658 TI - 5-(Tryptophyl)amino-1,3-dioxoperhydropyrido[1,2-c]pyrimidine-based potent and selective CCK(1)receptor antagonists: structure-activity relationship studies on the substituent at N2-position. AB - To establish structure-activity relationships a new series of analogues of the highly potent and selective CCK(1) receptor antagonist (4aS,5R)-2-benzyl-5-(N-Boc tryptophyl)amino-1,3-dioxoperhydropyrido[1,2-c]-pyrimidine (1a) modified at N2 position of the central scaffold has been prepared and evaluated as CCK receptor ligands. With this aim the N2-benzyl group has been replaced by methyl, cyclohexyl, aromatic groups, 1-phenylethyl, and 1-carboxy-2-phenylethyl group. Then, substituents with different electronic and steric properties were introduced into different positions of the phenyl group of analogues 19a and 19b. The results of the CCK receptor binding and in vitro functional activity evaluation suggest the importance of the lipophilic character and an appropriate spatial orientation of the moiety linked at the N2-position of the 1,3 dioxoperhydropyrido[1,2-c]pyrimidine template for potent and selective binding and antagonist activity at CCK(1) receptor subtype. The 2-cyclohexyl and (2S)-1 naphthyl derivatives 18a and (2S)-20a have emerged as more potent and selective CCK(1) receptor antagonists than the lead compound 1a. Additionally, the results confirm the (4aS,5R)-stereochemistry at the central bicyclic skeleton as an essential structural requirement for potent binding to this receptor subtype. PMID- 11405659 TI - Synthesis, nicotinic acetylcholine receptor binding, and antinociceptive properties of 2-exo-2-(2'-substituted 5'-pyridinyl)-7-azabicyclo[2.2.1]heptanes. Epibatidine analogues. AB - A convenient, high-yield synthesis of 7-tert-butoxycarbonyl-7 azabicyclo[2.2.1]hept-2-ene (5), which involved the addition of tributyltin hydride to 7-tert-butoxycarbonyl-2-p-toluenesulfonyl-7-azabicyclo[2.2.1]hept-2 ene (4) followed by elimination of the tributyltin and p-tolylsulfonyl groups using tetrabutylammonium fluoride was developed. The addition of 2-amino-5 iodopyridine to 5 under reductive Heck conditions provided 7-tert-butoxycarbonyl 2-exo-(2'-amino-5'-pyridinyl)-7-azabicyclo[2.2.1]heptane (6). Compound 6 was the key intermediate used to prepare epibatidine analogues where the 2'-chloro group on the pyridine ring was replaced with a fluorine (1b), bromine (1c), iodine (1d), hydroxy (1e), amino (1f), dimethylamino (1g), trifluoromethanesulfonate (1h), and hydrogen (1i) group. (+)- and (-)-Epibatidine and compounds 1b-d and 1i all possess similar binding affinities at the alpha(4)beta(2) nAChR receptors labeled by [(3)H]epibatidine. Compound 1f has affinity similar to nicotine, whereas compounds 1e, 1g, and 1h have much lower affinity. The binding affinity appears to be dependent upon the electronic nature of the substituent. However, other factors are also involved. None of the compounds possesses appreciable affinity for the alpha(7) nAChR labeled by [(125)I]iodo-MLA. With the exception of 1f and 1g, all the epibatidine analogues are full agonists (tail flick test) in producing antinociception after intrathecal injection in mice. PMID- 11405660 TI - Potent somatostatin undecapeptide agonists selective for somatostatin receptor 1 (sst1). AB - A family of analogues of des-AA(1,2,5)-[DTrp(8)/D2Nal(8)]-SRIF that contain a 4 (N-isopropyl)-aminomethylphenylalanine (IAmp) at position 9 was identified that has high affinity and selectivity for human somatostatin receptor subtype 1 (sst1). The binding affinities of des-AA(1,2,5)-[DTrp(8),IAmp(9)]-SRIF (c[H-Cys Lys-Phe-Phe-DTrp-IAmp-Thr-Phe-Thr-Ser-Cys-OH], CH-275) (7), des-AA(1,5) [Tyr(2),DTrp(8),IAmp(9)]-SRIF (CH-288) (16), des-AA(1,2,5) [Tyr(7),DTrp(8),IAmp(9)]-SRIF (23), and des-AA(1,2,5)-[DTrp(8),IAmp(9),Tyr(11)] SRIF (25) are about (1)/(7), (1)/(4), (1)/(125), and (1)/(4) that of SRIF-28 (1) to sst1, respectively, about (1)/(65), (1)/(130), <(1)/(1000), and <(1)/(150) that of 1 to sst3, respectively, and about or less than (1)/(1000) that of 1 to the other three human SRIF receptor subtypes. A substitution of DTrp(8) by D2Nal(8) in 7 to yield des-AA(1,2,5)-[D2Nal(8),IAmp(9)]-SRIF (13) and in 16 to yield des-AA(1,5)-[Tyr(2),D2Nal(8),IAmp(9)]-SRIF (17) was intended to increase chemical stability, selectivity, and affinity and resulted in two analogues that were less potent or equipotent with similar selectivity, respectively. Carbamoylation of the N-terminus as in des-AA(1,2,5)-[DTrp(8),IAmp(9),Tyr(11)] Cbm-SRIF (27) increased affinity slightly as well as improved selectivity. Monoiodination of 25 to yield 26 and of 27 to yield 28 resulted in an additional 4-fold increase in affinity at sst1. Desamination of the N-terminus of 17 to yield 18, on the other hand, resulted in significant loss of affinity. Attempts at reducing the size of the ring with maintenance of selectivity failed in that des-AA(1,4,5,13)-[Tyr(2),DTrp(8),IAmp(9)]-SRIF (33) and des-AA(1,4,5,6,12,13) [Tyr(2),DTrp(8),IAmp(9)]-SRIF (34) progressively lost affinity for all receptors. Both des-AA(1,2,5)-[DTrp(8),IAmp(9),Tyr(11)]-Cbm-SRIF (27) and des-AA(1,2,5) [DCys(3),DTrp(8),IAmp(9),Tyr(11)]-Cbm-SRIF (29) show agonistic activity in a cAMP assay; therefore, the structural basis for the agonist property of this family of analogues is not contingent upon the chirality of the Cys residue at position 3 as shown to be the case in 18-membered ring SRIF octapeptides. None of the high affinity structures described here showed receptor antagonism. We have prepared the radiolabeled des-AA(1,2,5)-[DTrp(8),IAmp(9),(125)ITyr(11)]-SRIF ((125)I-25) and des-AA(1,2,5)-[DTrp(8),IAmp(9), (125)ITyr(11)]-Cbm-SRIF ((125)I-27), used them as in vitro tracers, and found them to be superior to des-AA(1,5) [(125)ITyr(2),DTrp(8),IAmp(9)]-SRIF ((125)I-16) for the detection of sst1 tumors in receptor autoradiography studies. PMID- 11405661 TI - Characterization of melanocortin NDP-MSH agonist peptide fragments at the mouse central and peripheral melanocortin receptors. AB - The central melanocortin receptors, melanocortin-4 (MC4R) and melanocortin-3 (MC3R), are involved in the regulation of satiety and energy homeostasis. The MC4R in particular has become a pharmaceutical industry drug target due to its direct involvement in the regulation of food intake and its potential therapeutic application for the treatment of obesity-related diseases. The melanocortin receptors are stimulated by the native ligand, alpha-melanocyte stimulating hormone (alpha-MSH). The potent and enzymatically stable analogue NDP-MSH (Ac-Ser Tyr-Ser-Nle-Glu-His-DPhe-Arg-Trp-Gly-Lys-Pro-Val-NH(2)) is a lead peptide for the identification of melanocortin amino acids important for receptor molecular recognition and stimulation. We have synthesized nine peptide fragments of NDP MSH, deleting N- and C-terminal amino acids to determine the "minimally active" sequence of NDP-MSH. Additionally, five peptides were synthesized to study stereochemical inversion at the Phe 7 and Trp 9 positions in attempts to increase tetra- and tripeptide potencies. These peptide analogues were pharmacologically characterized at the mouse melanocortin MC1, MC3, MC4, and MC5 receptors. This study has identified the Ac-His-DPhe-Arg-Trp-NH(2) tetrapeptide as possessing 10 nM agonist activity at the brain MC4R. The tripeptide Ac-DPhe-Arg-Trp-NH(2) possessed micromolar agonist activities at the MC1R, MC4R, and MC5R but only slight stimulatory activity was observed at the MC3R (at up to 100 microM concentration). This study has also examined to importance of both N- and C terminal NDP-MSH amino acids at the different melanocortin receptors, providing information for drug design and identification of putative ligand-receptor interactions. PMID- 11405662 TI - Protease inhibitors: synthesis of a series of bacterial collagenase inhibitors of the sulfonyl amino acyl hydroxamate type. AB - A series of sulfonyl amino acyl hydroxamates incorporating alkyl/arylsulfonyl-N-2 nitrobenzyl-L-alanine was prepared. Related compounds were obtained by reaction of N-2-nitrobenzyl-L-Ala with aryl isocyanates, arylsulfonyl isocyanates, or benzoyl isothiocyanate, followed by the conversion of the COOH into the CONHOH moiety. The new compounds were assayed as inhibitors of the Clostridium histolyticum collagenase (ChC), a bacterial protease involved in the degradation of extracellular matrix. Many of the obtained hydroxamates proved to be effective bacterial collagenase inhibitors, the main contributor to activity being the substitution pattern at the sulfonamido moiety. The best ChC inhibitors were those containing pentafluorophenylsulfonyl and 3- and 4-protected aminophenylsulfonyl P(1)(') groups among others, with affinities in the low nanomolar range. This study also proves that the 2-nitrobenzyl- moiety, similarly to the 4-nitrobenyl one previously investigated (Scozzafava, A.; Supuran, C. T. J. Med. Chem. 2000, 43, 1858-1865) is an efficient P(2)(') anchoring moiety for obtaining potent bacterial collagenase inhibitors. PMID- 11405663 TI - Structural, CV and IR spectroscopic evidences for preorientation in PET-active phthalimido carboxylic acids. AB - Hydrogen bond and potassium cation mediated preorientation were detected for phthalimido acetic acid and the corresponding acetate. Evidence for these phenomena came from X-ray structure analysis as well as cyclic voltammetric and IR spectroscopic measurements. These interactions rationalize the photoinduced electron transfer (PET) reactivity of the substrates in photodecarboxylation reactions. PMID- 11405664 TI - A new receptor molecule for lysine and histidine in water: strong binding of basic amino acid esters by a macrocyclic host. AB - We present the new host molecule 1 which binds basic amino acid esters in water. It recognizes both positively charged groups of the amino acid esters by electrostatic and hydrogen bond interactions with its four strategically placed phosphonate anions. Selectivity for lysine is achieved by the correct distance between both bisphosphonate pairs. By contrast, the smaller amino acid esters arginine, ornithine, and histidine form 2:1 complexes with 1. In methanol, a double chelate assembly enforced by pi-cation interactions with the imidazolium cation leads to a very high association constant for the 1:histidine complex of 3 x 10(4) M(-)(1). PMID- 11405665 TI - Characterization of encapsulating supramolecules by using CSI-MS with ionization promoting reagents. AB - Guanidine nitrate, DMF, DMA, and DMSO are proven to be efficient ionization promoting reagents in coldspray ionization mass spectrometry (CSI-MS) for the self-assembled cage Pt compound 1 in aqueous solution. These reagents made it possible to characterize the structures of encapsulated guest molecules in the cage. Detailed structure determinations of the cage compound encapsulating various kinds of guest molecules by using positive CSI-MS with ionization promoting reagents are described. PMID- 11405666 TI - Remarkable alkali cation template effect in 1,5-bridged calix[8]arenes. AB - Treatment of 1,5-bridged calix[8]arenes 2-4 with alkali metal carbonates produces salts in which an included cation acts as a conformational template. This induces the folding of calix[8]arene skeleton in a conformation in which the triads of contiguous OH-bearing rings adopt a 3/4-cone conformation with all hydroxyls converging toward the same center. The template effect requires the presence of short bridges (ethylene, tetramethylene, diethylene glycol) and became stronger by increasing the cation dimension up to that of Cs(+). PMID- 11405667 TI - Unnatural natural products from the transannular cyclization of lathyrane diterpenes. AB - The potential of macrocyclic diterpenoids to afford natural product-like polycyclic compounds was demonstrated by the conversion of two lathyrane Euphorbia factors into a series of densely functionalized diterpenoids of unnatural skeletal type. Apparently, Nature is far from having fully exploited the built-in reactivity of these compounds to generate chemical diversity. PMID- 11405668 TI - Synthesis of 7beta-sulfur analogues of paclitaxel utilizing a novel epimerization of the 7alpha-thiol group. AB - Paclitaxel analogues with a sulfur group at the 7beta position were required for SAR studies. Attempts to generate these compounds by displacing a 7alpha leaving group with sulfur nucleophiles were unsuccessful. Instead, these compounds were successfully prepared from a 7beta-thiol intermediate that was obtained by a base catalyzed epimerization of the 7alpha-thiol derivative. The epimerization presumably proceeds through a thioaldehyde intermediate and exhibits the opposite stereochemical preference of its oxygen counterpart. PMID- 11405669 TI - Olefin metathesis in the design and synthesis of a globally constrained Grb2 SH2 domain inhibitor. AB - One drawback frequently associated with olefin metathesis-mediated peptide macrocyclization, the loss of side chain functionality at sites of ring closure, may be circumvented by incorporation of side chain functionality within the ring closing olefin segments. This approach is demonstrated in the preparation of a macrocyclic Grb2 SH2 domain antagonist designed as a conformationally constrained beta-bend mimic. PMID- 11405670 TI - An efficient synthesis of mimetics of neamine for RNA recognition. AB - As mimetics of neamine, several 4-heterocyclic 2-deoxystreptamine derivatives were chemically synthesized for RNA recognition. Conversion of 4-methylthiomethyl 5,6-di-O-acetyl-diazido-2-deoxystreptamine to the 4-chloromethyl derivative followed by reactions with different nuclophilic reagents gave the 4-heterocyclic 2-deoxystreptamine derivatives in satisfactory yields. PMID- 11405671 TI - Incorporation of stable organic radicals into cyclotriphosphazene: preparation and characterization of mono- and diradical adducts. AB - Stable cyclotriphosphazenes 4 and 5, incorporating one and two carbon radical centers, respectively, have been easily prepared and characterized. EPR spectroscopic studies in fluid solution at room temperature were carried out for both compounds and also for diradical 5 in frozen solvent matrixes. Spectral results are consistent with a triplet or degenerate singlet triplet ground state for 5. Reductive cyclic voltammetry shows a redox couple, being monoelectronic for 4 and bielectronic for 5. PMID- 11405672 TI - Electrophilic aromatic substitutions of silylated furans and thiophenes with retention of the organosilyl group. AB - The influence of trialkylsilyl groups on the nucleophilic reactivities of furans and thiophenes is determined by kinetic experiments. PMID- 11405673 TI - Nucleophilic reactivities of tributylstannyl-substituted furans and thiophenes. AB - Kinetic investigations of the reactions of benzhydryl cations with stannylated furans and thiophenes suggest that 2-(tributylstannyl)furan and -thiophene are preferentially attacked at the 5-position (k(rel), FcPhCH(+), 20 degrees C, CH(2)Cl(2)). PMID- 11405674 TI - In situ oxidation-imine formation-reduction routes from alcohols to amines. AB - Manganese dioxide is employed as an in situ oxidant for the one-pot conversion of alcohols into imines. In combination with polymer-supported cyanoborohydride (PSCBH), a one-pot oxidation-imine formation-reduction sequence is reported. This procedure enables alcohols to be converted directly into both secondary and tertiary amines. PMID- 11405675 TI - BINOL-salen metal catalysts incorporating a bifunctional design. AB - Salen metal complexes incorporating two chiral BINOL moieties have been synthesized and characterized crystallographically. The corresponding bisnaphthoxide complexes have been found to catalyze the asymmetric addition of benzyl malonate to cyclohexenone in up to 90% ee. With these modular catalysts, the Lewis acid and Bronsted base portions can be independently altered. PMID- 11405676 TI - Photoinduced electron transfer within porphyrin-containing poly(amide) dendrimers. AB - The synthesis and photophysical characterization of a series of free base porphyrin-containing polyamide dendrimers terminated with anthraquinone groups (FbP-Ga-AQ(n)(), a = 1-3, n = 12, 36, 108) are described. Substantial quenching (58-75%) of the porphyrin fluorescence of FbP-Ga-AQ(n)() is observed when compared to the analogous ethyl-terminated dendrimers (FbP-Ga-Et(n)()) in steady state fluorescence experiments and is attributed to intramolecular electron transfer. Time-resolved fluorescence experiments were fit to 2-3 exponentials, indicating multiple orientations for electron transfer, consistent with the flexible nature of these dendrimers. PMID- 11405677 TI - Exploring chemical diversity of epoxyquinoid natural products: synthesis and biological activity of (-)-jesterone and related molecules. AB - Enantioselective syntheses of the potent antifungal agent (-)-jesterone, its hydroxy epimer, and a dimeric quinone epoxide derivative are reported. The synthesis involves diastereoselective epoxidation of a chiral quinone monoketal derivative and regio- and stereoselective reduction of a quinone epoxide intermediate. PMID- 11405678 TI - Studies toward the total synthesis of scyphostatin: first entry to the highly functionalized cyclohexenone segment. AB - The cyclohexenone segment 2 of scyphostatin (1), a potent inhibitor of neutral sphingomyelinase, was synthesized in an enantioselective manner starting from the bromo ether 5 and D-serinal derivative 3. The synthetic method features a coupling reaction of 5 with 3 to construct the asymmetric quaternary carbon center and a stereospecific epoxide ring formation as the key steps. PMID- 11405679 TI - Unique fluorescence behavior of rhodamine B upon inclusion complexation with novel bis(beta-cyclodextrin-6-yl) 2,2'-bipyridine-4,4'-dicarboxylate. AB - Newly synthesized bis(beta-cyclodextrin-6-yl) 2,2'-bipyridine-4,4'-dicarboxylate was found to induce an unusual fluorescence enhancement of Rhodamine B (RhB) upon complexation. This effect is attributable to the equilibium shift of RhB to the highly fluorescent carboxylate ion form, which is induced by the cooperative binding by two appropriately preorganized cyclodextrin units in the bis(beta cyclodextrin). This sandwich complexation behavior was investigated by means of the fluorescence and 2D NMR spectroscopy. PMID- 11405680 TI - First evidence for the use of organosilver compounds in Pd-catalyzed coupling reactions; a mechanistic rationale for the Pd/Ag-catalyzed enyne synthesis? AB - Silver acetylides have been prepared and used in Pd-catalyzed coupling reactions. Enynes have thus been obtained in good yields. This work demonstrates that organosilver compounds could enter the Pd catalytic cycle; it also supports the role of silver acetylides as intermediates in the new Pd/Ag-catalyzed coupling reaction. PMID- 11405681 TI - A new C(1)-auxiliary for anomeric stereocontrol in the synthesis of alpha-sialyl glycosides. AB - The installation of the novel N,N-dimethylglycolamide ester auxiliary onto the C(1)-position of protected neuraminic acid donors allows for the exploitation of C(1)-neighboring group participation to generate sialoside conjugates with good to excellent alpha-selectivity under a variety of sialylation protocols, including those that would otherwise lead to nonselective or beta-selective sialoside products. PMID- 11405682 TI - Isolation and characterization of unsymmetrical C(60)Me(5)O(3)H, a cage-opened bisepoxide ketone: tautomerism involving a fullerene cage bond. AB - Bisepoxide ketone C(60)Me(5)O(3)H, possessing a nine-membered hole in the cage, has been isolated from the reaction of C(60)Cl(6) with methyllithium followed by hydrolysis. It is a tautomer of the recently isolated bisepoxide fullerenol, this tautomerism being the first example involving a cage C-C bond, and may be driven by cage strain. Like the fullerenol, the ketone gives a high C(58)(+) fragmentation ion intensity during EI mass spectrometry. PMID- 11405683 TI - A simple route to N-arylated 2-aminothiophenes as a new class of amorphous glass forming molecules. AB - By thermal decarboxylation of N-arylated 2-aminothiophene-5-cacrboxylates, a versatile, heavy-metal free method for preparing the title compounds as new class of highly reactive and easily oxidable, amorphous glass forming molecules has been elaborated. PMID- 11405684 TI - Regioselective palladium-catalyzed arylation of 2-furaldehyde. AB - An efficient regioselective method for the direct arylation of 2-furaldehyde to provide a range of pi-diverse 5-aryl-2-formylfuran derivatives is described. The method employs functionalized aryl halides and a catalytic amount of palladium(II) chloride under relatively mild conditions. PMID- 11405685 TI - A thioester ligation approach to amphipathic bicyclic peptide library. AB - An efficient approach to synthesize an amphipathic bicyclic peptide library from unprotected peptides is demonstrated through an on-resin intramolecular thioester ligation and an off-resin DMSO-mediated disulfide formation. PMID- 11405686 TI - Total synthesis of (-)-callystatin A. AB - An effective total synthesis of (-)-callystatin A (1), member of the leptomycin family of antibiotics, has been achieved. The synthesis features Evans extended aldol methodology to construct the northern polypropionate subunit and two separate Julia olefinations to assemble the conjugated dienes. The total synthesis proceeded in 2.3% overall yield with the longest linear sequence of 15 steps. PMID- 11405687 TI - Efficient syntheses of novel C2'-alkylated (+/-)-K252a analogues. AB - Recent efforts in our laboratories have resulted in a synthetic approach toward C2'-alkylated K252a analogues via extension of a K252a cyclofuranosylation strategy. The bis-indole-N-glycosidic coupling of 6-N-(3,4-dimethoxybenzyl) staurosporinone (21) with a number of highly functionalized carbohydrates has given access to previously unattainable, biologically relevant analogues. PMID- 11405688 TI - Formal [4+2]-annulation of chiral crotylsilanes: synthesis of the C19-C28 fragment of phorboxazoles. AB - A stereoselective synthesis of the C19-C28 fragment of phoborxazole A and B is described. The key step is an enantioselective [4 + 2]-annulation of a crotylsilane 10 with a propargylic aldehyde 11 affording a functionalized dihydropyran 12. A solvent-dependent stereoselective epoxidation of dihydropyrans is also documented. PMID- 11405689 TI - Generation and intermolecular reactions of 2-indolylacyl radicals. AB - The generation of 2-indolylacyl radicals from the corresponding phenyl selenoesters, aldehydes, and alpha-keto carboxylic acids and the scope of their participation in intermolecular addition reactions to carbon-carbon double bonds have been studied. Whereas the phenyl selenoester method has provided easy access to a variety of 1,4-dicarbonyl compounds bearing the 2-acylindole moiety, the glyoxylic acid route has been employed for the preparation of 2-indolyl pyridyl ketones. PMID- 11405690 TI - Highly enantioselective hydrogenation of (E)-beta-(acylamino)acrylates catalyzed by Rh(i)-complexes of electron-rich P-chirogenic diphosphines. AB - Excellent enantioselectivities up to 99.7% were achieved in the hydrogenation of (E)-beta-(acylamino)acrylates by the use of Rh(I)-complexes of electron-rich diphosphines, t-Bu-BisP and t-Bu-MiniPHOS. Low-temperature NMR experiments testify that monohydrides with beta-carbon atom of the substrate bound to rhodium are involved in the catalytic cycle. PMID- 11405691 TI - Preparation of 1,4-oxaselenin from agNO(3)/LDA-assisted reaction of 3-selena-4 pentyn-1-one as potential antitumor agents. AB - 1,4-Oxaselenins were synthesized from 3-selena-4-pentyn-1-ones by the use of AgNO(3) and LDA. One of the obtained oxaselenins, 2-(4-chlorophenyl)-6-phenyl-1,4 oxaselenin 5c, showed an inhibitory effect against the proliferation of human cancer cells and inducing effects on the early stage of apoptosis. PMID- 11405692 TI - Dehydrobenzoannulene-dimethyl-dihydropyrene hybrids: model systems for the synthesis of molecular aromatic probes. AB - Synthesis of two novel dehydrobenzoannulene-dimethyldihydropyrene (DBA-DDP) hybrids has been achieved using a Pd/Cu cross-coupling strategy. PMID- 11405693 TI - First total synthesis of (+/-)-taxifolial a and (+/-)-iso-caulerpenyne. AB - The first synthesis of (+/-)-taxifolial A and iso-caulerpenyne was accomplished. The key steps in the sequence are (1) the stereoselective assembly of a vinyltin derived from butynediol and a functionalized aldehyde and (2) the construction of the dienyne moiety via a Stille cross-coupling. PMID- 11405694 TI - Preparation and characterization of the fullerene diols 1,2-C(60)(OH)(2), 1,2 C(70)(OH)(2), and 5,6-C(70)(OH)(2). AB - The simple fullerene diols C(60)(OH)(2) and C(70)(OH)(2) were prepared by addition of RuO(4) followed by acid hydrolysis. The 1,2-C(60)(OH)(2) isomer was formed from C(60), and two isomers (1,2 and 5,6) of C(70)(OH)(2) were formed in the RuO(4) hydroxylation of C(70). These compounds are much more soluble in THF and dioxane than the parent fullerenes. More highly hydroxylated materials are formed as well. PMID- 11405695 TI - A rhodium carbene cyclization-cycloaddition cascade strategy toward the pseudolaric acids. AB - A rhodium carbene intramolecular cyclization-cycloaddition cascade was employed as the key reaction in the synthesis of the nucleus of the cytotoxic diterpenoids pseudolaric acids A and B. PMID- 11405696 TI - An alternative route for Carboni-Lindsey reaction: N,N cycloaddition of an alkene to s-tetrazine. AB - Quantum mechanical calculations show that N,N cycloaddition of alkenes and alkynes to s-tetrazines is possible as an alternative to the well-known C,C cycloaddition (Carboni-Lindsey reaction). Formation of 1,2,4-triazole derivatives (formal product of N,N cycloaddition) along with the pyrazole (formal product of C,C cycloaddition) corroborates this theoretical prediction. PMID- 11405697 TI - Anodic cyclization reactions: reversing the polarity of ketene dithioacetal groups. AB - Intramolecular coupling reactions of ketene dithioacetal groups with enol ether and alcohol nucleophiles have been studied. The reactions were initiated by an anodic oxidation of the ketene dithioacetal and proved to be compatible with the formation of five- or six-member rings, as well as the stereoselective generation of quaternary carbons. PMID- 11405698 TI - Formation of 1,4-disilyl-2-butenes from vinyl grignard reagent and chlorosilanes catalyzed by a titanocene complex. AB - Symmetrical 1,4-disilyl-2-butenes 1 have been prepared by the reaction of vinyl Grignard reagent with chlorosilanes. This reaction proceeds efficiently in the presence of a catalytic amount of titanocene dichloride at 0 degrees C in THF. When dichlorodiphenylsilane was used, 1,1-diphenyl-1-silacyclo-3-pentene 2 was obtained in a good yield. PMID- 11405699 TI - Tandem single-step construction of chiral hexahydrophenanthrenes: a concise route to (+)-ferruginol. AB - An efficient enantio- and diastereocontrolled construction of hydrophenanthrenes having either a quaternary or a tertiary benzylic stereogenic center has been developed by employing a tandem retro-aldol and intramolecular Friedel-Crafts alkylation sequence. Its application to a diastereocontrolled synthesis of an abietane diterpenoid (+)-ferruginol has also been demonstrated. PMID- 11405700 TI - Pseudo-C(3)-symmetric tertiary alcohol building block via group-selective hydroalumination: a synthesis of (-)-malyngolide. AB - The stereocontrolled preparation of tertiary alcohol 4 and its TMS surrogate 5, which share a pseudo-C(3)()-symmetry, is described. Compound 4 was used for the synthesis of (-)-malyngolide. PMID- 11405701 TI - Direct reductive amination of aldehydes and ketones using phenylsilane: catalysis by dibutyltin dichloride. AB - A procedure for direct reductive amination of aldehydes and ketones was developed which uses phenylsilane as a stoichiometric reductant and dibutyltin dichloride as a catalyst. Suitable amines included anilines and dialkylamines but not monoalkylamines. PMID- 11405702 TI - Sequential ring-closing metathesis and silicon-assisted cross-coupling reactions: stereocontrolled synthesis of highly substituted unsaturated alcohols. AB - A sequential ring-closing metathesis/silicon-assisted cross-coupling sequence has been developed. Alkenyldimethylsilyl ethers of omega-unsaturated alcohols undergo facile ring closure with Schrock's catalyst to afford five-, six-, and seven membered cycloalkenylsiloxanes bearing substituents on both alkenyl carbons. These siloxanes were highly effective coupling partners with various aryl and alkenyl halides and in the presence of Pd(0) afforded styrenes and dienes in high yield and specificity and with good functional group compatibility. PMID- 11405703 TI - A catalytic-enantioselective entry to planar chiral pi-complexes: enantioselective methoxycarbonylation of 1,2-dichlorobenzene-Cr(CO)(3). AB - The palladium-catalyzed mono-methoxycarbonylation of 1,2-dichlorobenzene tricarbonylchromium(0) has been achieved with up to 95% ee in the presence of the chiral ferrocene ligand (R,S)-PPF-pyrrolidine. It was found that the enantioselectivity strongly depends on the reaction time (conversion). Obviously, the initial enantioselectivity is enhanced by a subsequent kinetic resolution connected to the formation of the bis-methoxycarbonylated byproduct. PMID- 11405704 TI - Asymmetric synthesis of quaternary alpha-amino phosphonates using sulfinimines. AB - The addition of lithium diethylphosphonate to enantiopure ketosulfinimines is highly diastereoselective (>95%), affording the first examples of quaternary alpha-alkyl alpha-amino (arylmethyl)phosphonates. PMID- 11405705 TI - Total synthesis of (-)- and (+)-dysibetaine. AB - Glycidamides 6R and 6S were elaborated to (R,R)- and (S,S)-dysibetaines (1R and 1S) by intramolecular alkylation and functional group modification in 23% overall yield. The absolute stereochemistry of natural dysibetaine was established as S,S by comparison of the optical rotation of the natural product with that of the synthetic materials. PMID- 11405706 TI - Regioselective alkyl and alkynyl substitution reactions of epoxy alcohols by the use of organoaluminum ate complexes: regiochemical reversal of nucleophilic substitution reactions. AB - Unprecedented nucleophilic substitution reactions of 2,3-epoxy-1-alkanols with alkyl- and alkynylaluminum ate complexes have been studied and demonstrated to occur at the C2 position with extremely high stereoselectivity, i.e., with exactly reversed regioselectivity to that obtained in the substitution reactions by normal organoaluminum reagents, resulting in the formation of the C2-alkyl and C2-alkynyl substitution products in excellent yields. PMID- 11405707 TI - A new convergent route to conduritols A-F from a common chiral building block. AB - A diastereocontrolled route to conduritols A-F has been developed starting from a common chiral building block containing an oxabicyclo[3.2.1]octane framework. PMID- 11405708 TI - Kinetic resolution and double stereodifferentiation in catalytic asymmetric C-H activation of 2-substituted pyrrolidines. AB - Dirhodium tetrakis(S-(N-dodecylbenzenesulfonyl)prolinate) (Rh(2)(S-DOSP)(4)) catalyzed decomposition of methyl aryldiazoacetates in the presence of 2 substituted pyrrolidines results in highly diastereoselective and enantioselective C-H insertions. These reactions can proceed with impressive levels of double stereodifferentiation and kinetic resolution, which allows for three stereocenters to be controlled during the C-H insertion step. PMID- 11405709 TI - Stereospecific anionically promoted transannular hydride shifts in medium-ring hydroxy ketones. Probe of their reversibility and the potential for regiocontrol. AB - Examples are provided of stereospecific transannular oxidation-reduction processes involving the conjugate bases of delta-hydroxy ketones in a nine membered ring setting. The ability to control the direction of these equilibria by proper modulation of the solvent environment and level of hydroxyl group protection is demonstrated. MM3-derived steric energies of the isomer pairs suggest that the equilibrium distributions are the outcome of the extent to which intramolecular hydrogen bonding forces are disrupted by polar solvent molecules when present. PMID- 11405710 TI - Intramolecular photocycloaddition of cyclic thioimides as a method for heterocyclic synthesis. AB - Cyclic thioimides undergo photocycloaddition with tethered pi-bonds in a regiospecific manner to afford spirocyclic amidothietanes. These highly strained multicycles undergo a subsequent ring opening reaction to furnish novel fused pyrrolizinones. PMID- 11405711 TI - First enantioselective syntheses of (+)- and (-)-wilforonide by using chiral auxiliaries derived from the same chiral source. AB - [see structure]. The first enantioselective syntheses of both (+)-wilforonide (>98% ee) and (-)-wilforonide (>98% ee) have been accomplished by employing chiral auxiliaries derived from the same chiral source, (R)-pulegone. The bicyclic skeleton of wilforonide was constructed by using Mn(III)-based oxidative radical cyclization reactions of chiral beta-keto esters. The absolute configuration of natural wilforonide has been established to be (5aR,9aR). PMID- 11405712 TI - Solvent-dependent stereoselectivity in a Still-Wittig rearrangement: an experimental and ab initio study. AB - [see reaction]. The Still-Wittig rearrangement gave opposite selectivities for (Z:E)-alkenes in THF (3:1) vs toluene (1:3) in the synthesis of serine-proline dipeptide amide isosteres. Four transition states leading to (Z)-and (E)-alkenes with THF and without (representing toluene) were identified by ab initio calculations at the 3-21G* level. The calculated (Z:E)-ratios with THF (4.7:1) and without THF (1:3.2) suggested that the transition state geometries and energies were well-represented by the calculations. PMID- 11405713 TI - Constitution of Grignard reagent RMgCl in tetrahydrofuran. AB - The constitution of Grignard reagent, RMgCl (R = Me, tBu, Ph or benzyl), was investigated in the solid state by means of X-ray crystallography and in THF by coldspray ionization mass spectrometry (CSI-MS). Three types of crystal structures, (a) [Mg2(mu-Cl3)(THF)6](+)*[RMgCl2(THF)](-), (b) R2Mg4Cl6(THF)6, and (c) [2Mg2(mu-Cl3)(THF)6](+)*[R4Mg2Cl2]2-, were identified, and MeMg2(mu Cl3)(THF)4-6 were detected as major species of MeMgCl in solution. PMID- 11405714 TI - Quenching of biotinylated aequorin bioluminescence by dye-labeled avidin conjugates: application to homogeneous bioluminescence resonance energy transfer assays. AB - [see reaction]. Avidin conjugates containing the covalently attached dyes QSY-7 and dabcyl were prepared and shown to quench the bioluminescence of biotinylated aequorin. Quenching efficiency was shown to be dependent on both the label-to avidin ratio and the concentration of the avidin conjugate. These properties were exploited to develop a homogeneous bioluminescence resonance energy transfer (BRET) assay for biotin. PMID- 11405715 TI - Synthesis of the trisaccharide portion of the immunologic adjuvant QS-21A via sulfonium-mediated oxidative and dehydrative glycosylation. AB - [see structure]. The first synthesis of the trisaccharide fragment of the potent immunologic adjuvant QS-21A is reported. The key steps involve the application of sulfonium-mediated oxidative and dehydrative glycosidic couplings to construct the anomeric linkages in a short and convergent assembly of the branched trisaccharide. PMID- 11405716 TI - Synthesis of 5,10,15,20-tetrakis(4-tert- butyl-2,6-dicarboxyphenyl)porphyrin: a versatile bis-faced porphyrin synthon for D4-symmetric chiral porphyrins. AB - [see structure]. A versatile bis-faced porphyrin synthon, 5,10,15,20-tetrakis(4 tert-butyl-2,6-dicarboxyphenyl)porphyrin, was synthesized. The eight carboxyl groups were readily converted into various amide groups, and condensation with chiral amines led to various D4-symmetric chiral porphyrins with rigid structures. PMID- 11405717 TI - Anthraquinon-2-ylmethoxycarbonyl (Aqmoc): a new photochemically removable protecting group for alcohols. AB - [see structure]. Synthesis and photochemistry of a new photochemically removable protecting group for alcohols is described. Four carbonates of galactose derivatives (1-4) were synthesized from the corresponding arylmethanols via 4 nitrophenyl carbonate intermediates. Among them, photolysis of anthraquinon-2 ylmethoxycarbonyl (Aqmoc) galactose (1) proceeded with overall photolysis efficiency of 150 (quantum yield 0.10, and molar absorptivity 1500 M(-1) x cm( 1)) and rate constant of approximately 10(6) s(-1). To demonstrate its application to a biologically related molecule, 5'-Aqmoc-adenosine (5) was synthesized and photolyzed to yield adenosine in 91% yield. PMID- 11405718 TI - Rapid entry into the cryptophycin core via an acyl-beta-lactam macrolactonization: total synthesis of cryptophycin-24. AB - [see structure]. An efficient, concise approach to the macrolide core of the cryptophycins, potent antimitotic agents, has been achieved. The reaction sequence features a novel macrolactonization utilizing a reactive acyl-beta lactam intermediate that incorporates the beta-amino acid moiety within the 16 membered macrolide core. This highly modular approach, which allows for multiple alterations throughout the structure, was successfully applied to the total synthesis of cryptophycin-24. PMID- 11405719 TI - Total synthesis of (-)-salicylihalamide A. AB - [see structure]. A 16-step synthesis of the novel cytotoxin salicylihalamide A (1E) has been achieved in 3.3% overall yield using ring closing metathesis to generate the macrolide and addition of (1Z,3Z)-hexadienylcuprate (2), which was generated in situ from ethylcuprate and acetylene, to alkenyl isocyanate 3 to form the side chain. PMID- 11405720 TI - Two-component chiral phase transfer catalysts: enantioselective esterification of an N-acylated amino acid. AB - [see reaction]. The first example of a two-component chiral phase transfer catalyst is described which, operating in a biphasic solvent system, preferentially esterifies one enantiomer of a racemic N-acylated amino acid. The two-component catalyst is comprised of an achiral quaternary ammonium ion and a proline-derived chiral selector initially developed for the liquid chromatographic separation of enantiomers. PMID- 11405721 TI - Progress toward the total synthesis of kalihinane diterpenoids. AB - [see structure]. Studies toward the total synthesis of marine diterpenoids isolated from Acanthella sp., e.g., kalihinol A, are described. Efficient construction of the functionalized trans-decalin core (11) is achieved through intramolecular Diels-Alder cyclization followed by diastereoselective epoxidation and aziridination. PMID- 11405722 TI - Enantio- and diastereoselective reductive aldol reactions with iridium-pybox catalysts. AB - [see reaction]. A catalytic amount of [(cod)IrCl]2 and indane-pybox converts diethylmethylsilane, methyl acrylate, and certain aldehydes to the derived reductive aldol adduct with good enantio- and diastereocontrol. PMID- 11405723 TI - Digital communication through intermolecular fluorescence modulation. AB - [see reaction]. Ultraminiaturized processors incorporating molecular components can be developed only after devising efficient strategies to communicate signals at the molecular level. We have demonstrated that a three-state molecular switch responds to ultraviolet light, visible light, and H+, attenuating the emission intensity of a fluorescent probe. Intermolecular communication is responsible for the transduction of three input signals into a single optical output. The behavior of the communicating ensemble of molecules corresponds to that of a logic circuit incorporating seven gates. PMID- 11405724 TI - Development of a new triphase catalyst and its application to the epoxidation of allylic alcohols. AB - [see reaction]. A new triphase catalyst has been developed. When an aqueous solution of H3PW12O40 (1) was added to a solution of the amphiphilic chain copolymer 2, a new self-assembled and macroporous complex 3 was formed. This complex was effective as a catalyst in the epoxidation of allylic alcohols. Even in the use of 2.7 x 10(-5) mole equiv of the catalyst, the epoxidation with aqueous H2O2 proceeded without organic solvents to give the corresponding epoxy alcohols in high yields. PMID- 11405725 TI - Dithiane- and trithiane-based photolabile scaffolds for molecular recognition. AB - [see structure]. A modular synthetic approach to novel dithiane- and trithiane based photolabile molecular hosts equipped with elements of molecular recognition is developed. The approach provides ready access to a family of amino-derivatized photocleavable molecular systems capable of hydrogen-bonding-based recognition of biologically relevant molecules, e.g., ureas, barbiturates etc. These systems undergo efficient photofragmentation in the presence of external (e.g., benzophenone) or internal (e.g., nitropyridine) electron-transfer sensitizers. PMID- 11405726 TI - A novel [60]fullerene amino acid for use in solid-phase peptide synthesis. AB - [see structure]. A fullerene derivative containing a free amino group has been condensed with N-Fmoc-L-glutamic acid alpha-tert-butyl ester to give a C60 functionalized amino acid. The carboxylic end of this amino acid has been deprotected in acidic conditions, and the resulting acid has been used for solid phase peptide synthesis. The final peptide, cleaved from the resin, was very soluble in water solutions and showed antimicrobial activity against two representative bacteria. PMID- 11405727 TI - Synthesis of beta-lactams from diazoketones and imines: the use of microwave irradiation. AB - [see reaction]. The transformation of diazoketones derived from alpha-amino acids to ketenes that, in turn, react further with imines to afford beta-lactams, can be realized not only by utilizing photochemical reaction conditions but also under the action of microwave irradiation. Under the latter reaction conditions 4 alkenyl-substituted beta-lactams derived from amino acids, substrates that were not previously accessible, have been prepared. beta-Lactams possessing a trans substitution pattern at the ring were obtained exclusively. PMID- 11405728 TI - Triethylborane-induced radical reactions with gallium hydride reagent HGaCl2. AB - [see reaction]. A gallium hydride reagent, HGaCl2, was found to act as a radical mediator, like tributyltin hydride. Treatment of alkyl halides with the gallium hydride reagent, generated from gallium trichloride and sodium bis(2 methoxyethoxy)aluminum hydride, provided the corresponding reduced products in excellent yields. Radical cyclization of halo acetals was also successful with not only the stoichiometric gallium reagent but also a catalytic amount of gallium trichloride combined with stoichiometric aluminum hydride as a hydride source. PMID- 11405729 TI - New ligands bearing chiral bioactive fragments. AB - [see reaction]. Reliable and practical synthetic routes for the construction of hybrid molecules bearing both a chelating center and a useful biofunction are presented. They comprise the sequential cross-coupling reaction between ethynylated synthons with iodo-substituted L-tyrosine derivatives and provide access to various rationally designed chiral ligands. PMID- 11405730 TI - Silylcupration of 1,3-dienes followed by an electrophilic trapping reaction. AB - [see reaction]. Silylcupration reaction of 1,3-dienes with a cyanocuprate reagent, PhMe2SiCuCNLi, followed by an electrophilic trapping has been reported for the first time. The use of allylic phosphates as electrophiles resulted in a highly regioselective reaction with overall 1,4-addition of the silyl and allyl moieties across the diene. PMID- 11405731 TI - Grignard reactions of 4-substituted-2-keto-1,3-dioxanes: highly diastereoselective additions controlled by a remote alkyl group. AB - [see reaction]. The reactions of Grignard reagents with a representative series of simple cis-2-keto-4-substituted-1,3-dioxanes have been investigated. The stereochemical outcome of these highly diastereoselective additions (dr > 90:10) is consonant with Cram's chelate model on the assumption that RMgX coordinates preferentially with the ring oxygen remote from the C(4) substituent. PMID- 11405732 TI - Efficient scavenging of Ph3P and Ph3P=O with high-loading merrifield resin. AB - [see reaction]. A simple, highly effective method for removing triphenylphosphine and/or triphenylphosphine oxide from reaction mixtures is described. Commercially available high-loading chloromethylated polystyrene 1, modified in situ with NaI, acts as a scavenger resin. Several coupling reactions catalyzed by Pd(0) or Ni(0) which require the removal of triphenylphosphine are tested. PMID- 11405733 TI - Highly selective one-pot synthesis of spirophosphoranes exhibiting reversed apicophilicity by oxidation of dianions generated from P-H spirophosphorane. AB - [see reaction]. Mild and highly selective one-pot procedures for obtaining phosphoranes that exhibit reversed (O-cis) apicophilicity are described. On the basis of the procedures, O-cis phosphorane bearing an aryl group (R = 2,4,6-tri-i propylphenyl) could be isolated for the first time; the procedure is also applicable for alkyl derivatives. Particularly effective was the use of I2 as an oxidizing reagent. PMID- 11405734 TI - Synthesis of conjugated diacetylene, metal-chelating monomers for polymerizable monolayer assemblies. AB - [see structure]. Self-assembled monolayers (SAMs) of thiols on gold have been used for numerous applications. For protein targeting applications, one successful strategy is to use a metal-chelating SAM. It has also been demonstrated that polymerized SAMs are much more stable than non-polymerized counterparts. We report herein, the synthesis of several polymerizable, metal chelating thiols capable of complexing luminescent lanthanide ions. PMID- 11405735 TI - Biradical and zwitterionic cyclizations of oxy-substituted enyne-allenes. AB - [see reaction]. Oxyanion substitution of enyne-allenes causes both Myers-Saito and Schmittel cyclizations to switch their product formation preferences from diradicals to polar, closed-shell singlets. The oxyanion stabilization is larger for the Schmittel products than the Myers-Saito products because the latter must sacrifice aromaticity to maximize interaction. The changing character of the different reaction paths is reflected in their activation energies. PMID- 11405736 TI - Photochemical ring opening of 7-benzoyl- and 7 methoxycarbonyldibenzonorcaradienes. competing 1,2-hydrogen shift and cyclization reactions of 1,3-diradicals. AB - [see reaction]. The UV irradiation of dibenzonorcaradienes bearing an acyl or alkoxycarbonyl substituent in the 7-position results in formation of substituted phenanthrenes, as well as cis-trans isomerization of the starting material. This reaction apparently proceeds via intermediate formation of a short-lived (tau = 1 20 ns) 1,3-diradical, which is produced by photochemical cleavage of one cyclopropane bond, while no evidence of alpha-carbonylcarbene formation was found. PMID- 11405737 TI - A configurationally stable alkoxy allenyl zinc reagent, en route to anti-anti vicinal amino diols. AB - [see reaction]. The reaction of an alkoxyallenyl zinc reagent with benzyl imines derived from lactic and mandelic acids proceeds highly diastereoselectively and leads to 2-amino-1,3-diol derivatives with an anti-anti pattern. PMID- 11405738 TI - Synthesis of new cationic lipids from an unsaturated glycoside scaffold. AB - [see structure]. We report the synthesis of new cationic lipids. These amphiphiles present a hydrophobic domain connected to a guanidinium entity by an unsaturated glycoside scaffold. The synthetic strategy using amide or acetal linkage led to various mono- and bicatenar derivatives. Investigation of their physicochemical properties indicated that these new compounds compact DNA. PMID- 11405739 TI - Catalytic asymmetric vinylation of ketone enolates. AB - [see reaction]. A protocol for the catalytic asymmetric vinylation of ketone enolates has been developed. Key to the success of this process was the development of new electron-rich chiral monodentate ligands. PMID- 11405740 TI - Tandem conjugate reduction-aldol cyclization using Stryker's reagent. AB - [see reaction]. Conjugate reduction by Stryker's reagent to form copper enolates, followed by intramolecular aldol cyclization, successfully generated five- and six-membered carbocycles in one pot efficiently. This tandem reaction is generally diastereoselective and provides good yields of the beta-hydroxyketones without any dehydration at low temperatures. PMID- 11405741 TI - Alkynoates as a source of reactive alkylinides for aldehyde addition reactions. AB - [see reaction]. The reaction of activated alkynes with carbonyl compounds in the presence of a catalytic amount of a nucleophile leads to enol-protected functionalized propargyl alcohols and 1,3-dioxolane compounds by way of a mild carbon-carbon bond formation reaction. PMID- 11405742 TI - Stereoselective synthesis of iso-dolaproine via dynamic kinetic resolution. AB - [see reaction]. An efficient multigram-scale synthesis of optically pure Boc (2S,3R,4S)-iso-dolaproine is reported using dynamic kinetic resolution (DKR). The catalytic asymmetric hydrogenation of ethyl (4S)-3-(2'-pyrrolidinyl)-3-oxo-2 methyl propanoate hydrochloride using in situ generated Ru[(S)-MeO-BIPHEP]Br2 catalyst affords the anti beta-hydroxy alpha-methyl ester quantitatively. The two new stereogenic centers are simultaneously controlled with high diastereoselectivity. PMID- 11405743 TI - A new and general synthetic pathway to strychnos indole alkaloids: total syntheses of (-)-dehydrotubifoline and (-)-tubifoline by palladium-catalyzed asymmetric allylic substitution. AB - [see reaction]. A novel procedure for the synthesis of an indole skeleton was developed. Treatment of a cyclohexenol derivative having a silyloxymethyl group at the 2-position with N-tosyl-o-bromoaniline in the presence of Pd2dba3*CHCl3 and (S)-BINAPO gave compound 6a with 84% ee in 75% yield. Compound 6a was converted into 11, which was treated with Pd(OAc)2 and Me(2)PPh in the presence of Ag2CO3 to give indoline derivative 12. From 12, we succeeded in the total syntheses of (-)-dehydrotubifoline and (-)-tubifoline. PMID- 11405744 TI - Generation and reaction of alkene radical cations under nonoxidizing conditions: synthesis of the pyrrolizidine nucleus. AB - [see reaction]. Stable beta-phosphatoxy nitroalkanes, readily assembled by the Henry reaction and subsequent phosphorylation, serve as good precursors to alkene radical cations on treatment with triphenyltin or tributyl hydride and AIBN in benzene at reflux. When the beta-phosphatoxy nitroalkane is suitably functionalized with nucleophilic groups, substitutions can be achieved with the formation of heterocyclic rings. When the nucleophile is an allylamine, tandem processes occur giving pyrrolizidines. PMID- 11405745 TI - Solid state NMR of a molecular self-assembly: multinuclear approach to the cyanuric acid-melamine system. AB - [see structure]. Trinuclear MAS NMR, involving naturally abundant (13C, 15N) and easily deuterated (2H) nuclei, is shown to offer newer opportunities to study molecular self-assembly in noncrystalline supramolecular systems. PMID- 11405746 TI - Total synthesis of (+)-arteanniun M using the tandem oxy-Cope/ene reaction. AB - [see reaction]. The first total synthesis of (+)-arteannuin M was accomplished using the tandem oxy-Cope/transannular ene reaction as the key step to construct the bicyclic core of the natural product. The tandem reaction proceeded with high diastereo- and enantiomeric excess. PMID- 11405747 TI - Enantioselective epoxidation of terminal olefins by chiral dioxirane. AB - [see reaction]. This paper describes an enantioselective epoxidation of terminal olefins using chiral ketone 3 as catalyst and Oxone as oxidant. Up to 85% ee has been obtained. PMID- 11405748 TI - 25,27-dithiasapphyrin and pyrrole-inverted isomer of 21,23-dithiaporphyrin from condensation of pyrrole and 2,5-bis(p-tolylhydroxymethyl)thiophene. AB - [see structure]. A novel aromatic isomer of 5,10,15,20-tetra(p-tolyl)-21,23 dithiaporphyrin (S2TTP) with an inverted pyrrole ring, 5,10,15,20-tetra(p-tolyl) 2-aza-21-carba-22,24-dithiaporphyrin (S2CTTP), and 5,10,15,20-tetra(p-tolyl) 25,27-dithiasapphyrin (25,27-S2TTSH), have been obtained by a condensation of 2,5 bis((p-tolyl)hydroxymethyl)thiophene and pyrrole. A conformational equilibrium, unique in a sapphyrin class, between two S(27)-thiophene-flipped and planar structures of neutral 25,27-S2TTSH was detected by 1H NMR. PMID- 11405749 TI - Optically active beta-ketoiminato cationic cobalt(III) complexes: efficient catalysts for enantioselective carbonyl-ene reaction of glyoxal derivatives. AB - [see reaction]. Optically active beta-ketoiminato cationic cobalt(III) complexes were synthesized as effective Lewis acid catalysts for the enantioselective carbonyl-ene reaction. In the presence of a catalytic amount of cobalt(III) hexafluoroantimonate derived from the optically active 1,2-diphenyl-1,2 ethanediamine, the carbonyl-ene reaction with a variety of alkenes and glyoxal derivatives smoothly proceeded to afford the corresponding homoallylic alcohols in good-to-high yields with high enantioselectivities. PMID- 11405750 TI - A new approach to 4-alkylthio-1,3-dithiole-2-thione: an unusual reaction of a zinc complex of 1,3-dithole-2-thione-4,5-dithiolate. AB - [see reaction]. A new and facile approach to 4-alkylthio-1,3-dithiole-2-thione starting from easily accessible reactants was described. This approach was based on the unusual reaction of a zinc complex of 1,3-dithiole-2-thione-4,5-dithiolate with electrophilic reagents in the presence of 3-picolyl chloride hydrochloride/or 4-picolyl chloride hydrochloride/or pyridine hydrochloride. PMID- 11405751 TI - Amination of grignard reagents with retention of configuration. AB - [see reaction]. The stereochemistry of electrophilic amination has been probed using the chiral Grignard reagent 5, in which the magnesium-bearing carbon atom is the sole stereogenic center. Amination with azidomethyl phenyl sulfide 1 and with O-sulfonyloxime 2 were found to proceed with full retention of configuration. PMID- 11405752 TI - Studies on the synthesis of pectenotoxin II: synthesis of a C(11)-C(26) fragment precursor via [3 + 2]-annulation reactions of chiral allylsilanes. AB - [see reaction]. A synthesis of tetracycle 2 corresponding to the C(11)-C(26) fragment of pectenotoxin II is described. The synthesis features two highly stereoselective [3 + 2]-annulation reactions of chiral allylsilanes, generated via allylboration of aldehydes with the chiral gamma-silylallylborane 4 or the gamma-silylallylboronate 19, for construction of the highly substituted C and E rings. PMID- 11405753 TI - Formal synthesis of uvaricin via palladium-mediated double cyclization. AB - [see reaction]. A novel palladium-mediated, ligand-controlled double cyclization is reported. The C2-symmetric diene produced was desymmetrized via Sharpless asymmetric dihydroxylation and was further transformed into a known intermediate in the synthesis of uvaricin. PMID- 11405754 TI - Trypanosome and animal lanosterol synthases use different catalytic motifs. AB - [see reaction]. Animals, fungi, and some protozoa convert oxidosqualene to lanosterol in the ring-forming reaction in sterol biosynthesis. The Trypanosoma cruzi lanosterol synthase has now been cloned. The sequence shares with the T. brucei lanosterol synthase a tyrosine substitution for the catalytically important active-site threonine found in animal and fungal lanosterol synthases. PMID- 11405755 TI - Toward globular macromolecules with functionalized interiors: design and synthesis of dendrons with an interesting twist. AB - [see reaction]. Design and synthesis of a novel class of monodendrons, in which the functional units can potentially be directed toward the concave interiors of dendrimers, are described. The key feature of the design is the placement of the amphiphilic and the AB2 functional groups in orthogonal planes. PMID- 11405756 TI - Topical imiquimod to treat intraepidermal carcinoma. PMID- 11405757 TI - The rediscovery of previously described dermatoses. PMID- 11405759 TI - Nodular fasciitis. PMID- 11405760 TI - Fatty acid analysis of transplanted adipose tissue. AB - OBJECTIVE: To determine whether autologously transferred human adipose tissue maintains viability in vivo for prolonged periods. DESIGN: Six healthy female patients (mean age, 61.5 years; mean body mass index, 23.4 kg/m2) received autologous fat transplants from the gluteus to the nasolabial folds. Subcutaneous fat was sampled from facial and gluteal sites 4 times in 1 year. SETTING: Private practice, basic science research center. INTERVENTION: After local anesthesia, 10 g of subcutaneous adipose tissue was harvested from the right buttock of each patient. Ten milligrams of adipose tissue was aspirated from the right nasolabial fold. Five grams of gluteal fat was then injected into each nasolabial fold using a uniform monolayer threading technique with no overcorrection. As controls, 10 mg of adipose tissue was obtained from the opposite left buttock and left cheek. Adipose tissue from the transplanted and control facial and gluteal sites was sampled at 4, 6, and 12 months after transplantation. MAIN OUTCOME MEASUREMENTS: Gluteal fat has more monounsaturated fatty acids and less saturated fatty acids than facial fat. This unique site-specific fatty acid pattern was used to assess the course of the survival of transplanted adipose tissue in the nasolabial region. In all fat samples, the percent area (weight percentage) was obtained for each fatty acid (C12:0 to C22:6 omega-3) using capillary gas chromatography. Clinical results were also analyzed by macrophotographs. RESULTS: As expected, gluteal fat had significantly more monounsaturated fatty acids and less saturated fatty acids than facial fat. In 5 of 6 patients, at 4, 6, and 12 months after transplantation, the fatty acid pattern at the transplanted recipient site was similar to the pattern of the control facial site. However, at 4 months, 1 patient had a fatty acid pattern in the transplant recipient site that was similar to the pattern of her gluteal fat. This pattern persisted for 1 year. Fat retention at the transplant site was corroborated by photographic assessment. CONCLUSIONS: Long-term adipocyte survival is an achievable goal following fat transfer. The importance of harvesting and injection techniques as well as adipose tissue characteristics require further study. PMID- 11405761 TI - Incidence of cancer among patients with hidradenitis suppurativa. AB - BACKGROUND: On the basis of some case reports, a relationship has been suggested between hidradenitis suppurativa (HS) and the development of nonmelanoma skin cancer. OBJECTIVES: To confirm this relationship and to explore the risk of other cancers among patients with HS. PATIENTS: Patients with a discharge diagnosis of HS were obtained from the computerized database of hospital discharge diagnoses from January 1, 1965, through December 31, 1997. A total of 2119 patients with HS were identified. SETTING: All hospitals in Sweden. DESIGN: With record linkage to the Swedish National Cancer Registry, standardized incidence ratios (SIR [the ratio of the observed to expected incidence]) were calculated to estimate relative risk. RESULTS: The risk of developing any cancer in the cohort with HS increased 50% (95% confidence interval of SIR, 1.1-1.8, based on 73 observed cases). Statistically significant risk elevations were observed for nonmelanoma skin cancer (5 cases; SIR, 4.6; 95% confidence interval, 1.5-10.7), buccal cancer (5 cases; SIR, 5.5; 95% confidence interval, 1.8-12.9), and primary liver cancer (3 cases; SIR, 10.0; 95% confidence interval, 2.1-29.2). CONCLUSIONS: This study confirms an increased risk of nonmelanoma skin cancer among patients with HS. The risk for buccal cancer and primary liver cancer was also elevated among this cohort, but these associations should be interpreted cautiously because the combination of multiple significance testing and the few observed cases may have generated chance findings. PMID- 11405762 TI - Detection of IgA autoantibodies to desmogleins by an enzyme-linked immunosorbent assay: the presence of new minor subtypes of IgA pemphigus. AB - OBJECTIVE: To examine the frequency of antidesmoglein 1 (Dsg1) and antidesmoglein 3 (Dsg3) IgA autoantibodies in IgA pemphigus. DESIGN: We developed an enzyme linked immunosorbent assay against recombinant Dsg1 and Dsg3 to detect IgA autoantibodies. PATIENTS: Twenty-two patients with IgA pemphigus were studied. Among them, 10 patients had subcorneal pustular dermatosis type, 9 patients had intraepidermal neutrophilic IgA dermatosis type, and 3 patients had pemphigus foliaceus-like clinical features. RESULTS: Of the 22 cases of IgA pemphigus, 3 cases were positive for anti-Dsg1 IgA antibodies and only 1 case was positive for anti-Dsg3 IgA antibodies. In those 4 cases, there were no IgA autoantibodies against other components of the keratinocyte cell surfaces because preincubation with the respective recombinant desmogleins removed the immunoreactivity on immunofluorescence. All 10 patients with subcorneal pustular dermatosis type IgA pemphigus were positive against desmocollin 1 expressed on COS-7 cells. No target antigen was detected in the other 8 cases. CONCLUSIONS: Desmogleins were recognized by IgA antibodies of a few patients with IgA pemphigus. Considering that subcorneal pustular dermatosis type IgA pemphigus recognizes desmocollin 1, autoimmune targets of IgA pemphigus are more heterogeneous than previously considered. PMID- 11405763 TI - Imaging blood flow in human port-wine stain in situ and in real time using optical Doppler tomography. AB - BACKGROUND: Optical Doppler tomography (ODT) combines laser Doppler flowmetry with optical coherence tomography to obtain high-resolution images of blood flow in human skin in situ and in real time. OBSERVATIONS: We present a case in which ODT was used on a patient with a port-wine stain (PWS) birthmark to document the change of blood flow in response to laser therapy. It might be possible to use ODT blood flow measurements in situ to assist in assessing the efficacy of laser PWS therapy. If partial restoration of flow occurs immediately or shortly after laser exposure, indicative of reperfusion due to inadequate blood vessel injury, the PWS can be retreated using higher light dosages. Retreatment is continued until the measured Doppler shift is zero due to a permanent reduction in blood flow, indicative of irreversible microthrombus formation in the PWS vessels. CONCLUSIONS: We have demonstrated that ODT may be used for noninvasive imaging of blood vessels in PWS skin. Moreover, ODT will potentially allow laser therapy to be optimized on an individual patient basis by providing a fast, semiquantitative evaluation of the efficacy of PWS laser therapy in situ and in real time. PMID- 11405764 TI - Significant absorption of topical tacrolimus in 3 patients with Netherton syndrome. AB - BACKGROUND: Tacrolimus is a macrolide immunosuppressant approved in oral and intravenous formulations for primary immunosuppression in liver and kidney transplantation. Topical 0.1% tacrolimus ointment has recently been shown to be effective in atopic dermatitis for children as young as 2 years of age, with minimal systemic absorption. We describe 3 patients treated with topical 0.1% tacrolimus who developed significant systemic absorption. OBSERVATION: Three patients previously diagnosed as having Netherton syndrome were treated at different centers with 0.1% tacrolimus ointment twice daily. Two patients showed dramatic improvement. All patients were found to have tacrolimus blood levels within or above the established therapeutic trough range for oral tacrolimus in organ transplant recipients. None of these patients developed signs or symptoms of toxic effects of tacrolimus. CONCLUSIONS: Patients with Netherton syndrome have a skin barrier dysfunction that puts them at risk for increased percutaneous absorption. The Food and Drug Administration recently approved 0.1% tacrolimus ointment for the treatment of atopic dermatitis. Children with Netherton syndrome may be misdiagnosed as having atopic dermatitis. These children are at risk for marked systemic absorption and associated toxic effects. If topical tacrolimus is used in this setting, monitoring of serum tacrolimus levels is essential. PMID- 11405765 TI - Discordance between facial wrinkling and the presence of basal cell carcinoma. AB - BACKGROUND: During routine surgical treatment of basal cell carcinomas (BCCs), we observed an apparent inverse relationship between the presence of a BCC and significant wrinkling of the face. To ascertain the veracity of this observation, we performed a clinical and questionnaire-based case-referent study. OBSERVATION: One hundred eighteen successive white patients (mean +/- SD age, 71.9 +/- 9.5 years) attending the hospital for surgical treatment of a BCC and 121 control (no skin cancer) patients (mean +/- SD age, 69.1 +/- 10.8 years) were assessed for grade of facial wrinkling using a previously validated photonumeric scale of photoaging and completed a questionnaire about sun exposure. Despite being older (P =.03), patients with a BCC were found to have a lower mean grade of wrinkling than controls (P =.001). Using logistic regression, increasing grade of wrinkling was associated with a progressive reduction in risk of developing a BCC. CONCLUSION: Mechanisms responsible for the production of facial wrinkles may either be separate to or mitigate against the development of a BCC of the face. PMID- 11405766 TI - Penile pemphigus. AB - BACKGROUND: Penile skin involvement in patients with pemphigus vulgaris has been rarely reported. This study describes the involvement of penile skin in 12 patients with pemphigus vulgaris. OBSERVATIONS: Of the 12 patients, 10 had involvement of the skin and mucous membranes. Two patients had involvement of the oral mucosa only and no cutaneous involvement. None of the patients had urethral involvement. We did not observe isolated involvement of penile skin only, in the absence of disease elsewhere. Using monkey esophagus as substrate, all the patients had detectable levels of antibodies to keratinocyte cell surface antigen(s) in their serum samples. Since histological, serological, and clinical evidence of pemphigus was present, biopsies of the penile skin were not done. Topical therapy was concomitantly used with systemic therapy. Once treated and resolved, recurrence of penile lesions was not observed during the long-term follow-up. CONCLUSIONS: Involvement of penile skin is rare and was observed with the presence of pemphigus lesions in other areas of the body. Lesions involving the penile skin were most commonly seen on the glans. No sequelae or functional abnormalities were observed on long-term follow-up. PMID- 11405767 TI - Vesiculopustular eruptions in Down syndrome neonates with myeloproliferative disorders. AB - BACKGROUND: Infants with Down syndrome are at increased risk for hematologic abnormalities, including leukemoid reaction, transient myeloproliferative disorder, and congenital leukemia. The differential diagnosis of a vesiculopustular eruption in an infant with Down syndrome and these hematologic abnormalities is broad and includes benign, self-limited disorders as well as life-threatening infections. OBSERVATION: We describe 3 newborns with Down syndrome and vesiculopustular eruptions associated with myeloproliferative disorders during the neonatal period. These lesions differ from other neonatal vesicular eruptions in that they have a unique distribution, display pathergy, and contain immature hematopoietic cells similar to circulating blast cells. Resolution occurs without treatment as the hematologic disorder subsides. CONCLUSIONS: Infants with Down syndrome and hematologic abnormalities may have a cutaneous eruption that has characteristic clinical and histopathologic findings. It is possible that this eruption has been unrecognized in the past because of its self-limited course. Whether this eruption is a prognostic factor for the subsequent development of leukemia is uncertain. PMID- 11405768 TI - Rates of cutaneous reactions to drugs. AB - OBJECTIVE: To determine the validity, magnitude, precision, and applicability of data on the rates at which drugs cause adverse cutaneous reactions. DESIGN: Systematic review of the medical literature. DATA SOURCE: The MEDLINE database was searched (1966-August 2000) for studies that contain information on the rates of cutaneous reactions to drugs. The bibliographies of retrieved articles and review articles were also examined to find relevant studies. MAIN OUTCOME MEASURES: Studies that included primary data on cutaneous reaction rates to drugs were evaluated for their validity, magnitude, precision, and applicability, using guidelines derived from existing guidelines for the evaluation of articles about harm and prognosis. RESULTS: Nine studies met the study criteria. Five of the studies were based on prospectively collected data on medical inpatients, 2 were retrospective studies based on chart or computerized medical record review, and 2 were based on spontaneous reports and consumption data. The morbilliform drug exanthem and urticaria were the most common cutaneous reactions to drugs. Reaction rates varied from 0% to 8% and were highest for antibiotics (in the range of 1% to 8% for several classes of antibiotics). CONCLUSIONS: Despite differences in the methods of the studies reviewed and their time of execution, there is remarkable agreement in the results. Reaction rates (and 95% confidence intervals) are available for many commonly used drugs. PMID- 11405769 TI - What proportion of dermatological patients receive evidence-based treatment? AB - OBJECTIVE: To determine the proportion of dermatological patients who are offered evidence-based therapy in the routine dermatological practice. METHODS: For every patient seen for the first time at one of our tertiary hospital setting clinics between April and May 1999, the primary diagnosis and the primary intervention were recorded. For each primary diagnosis-primary intervention combination, evidence was searched for in electronic databases from January 1966 to December 1999. The proportion of patients who were offered evidence-based interventions was calculated as the main outcome measure. RESULTS: With a study sample of 136 patients, 61 different diagnosis-treatment couples were generated and 94 queries on electronic databases were performed (to account for "primary interventions" including more than 1 drug or treatment modality). Eighty-seven (64%) of 136 patients received evidence-based interventions. Evidence from randomized controlled trials was found for 69 patients (50.7% of the sample). Controlled studies lacking randomization or double blinding or including fewer than 20 patients per treatment group dealt with treatments offered to 14 patients (10.3%). The treatments offered to 4 patients (2.9%) were judged to have self evident validity (ie, trials unanimously judged unnecessary). Symptomatic and supportive measures accounted for most interventions lacking substantial evidence (36% of the patients), but we had to include in this class other important treatment regimens, mainly for rare conditions. CONCLUSIONS: Most of the study patients received evidence-based care. However, published trials should be carefully appraised, and relevance of clinical end points should be evaluated together with methodological issues. More accessible, clinically oriented, evidence-based information sources are needed. PMID- 11405770 TI - The risk of malignancy associated with psoriasis. AB - OBJECTIVE: To measure the incidence of cancer in patients with psoriasis, stratified by the severity of their disease. DESIGN: A cohort study. SETTING: Administrative claims records obtained from Medicaid programs in 3 US states. PARTICIPANTS: All individuals in the claims database who qualified for 1 of the 5 following groups: severe psoriasis as defined by treatment with systemic medication, less severe psoriasis, severe eczema, history of organ transplantation, and hypertension. MAIN OUTCOME MEASURE: A diagnosis of cancer. RESULTS: Individuals with severe psoriasis were more likely to develop a malignancy than those with hypertension (risk ratio, 1.78; 95% confidence interval [CI], 1.32-2.40). The risk of malignancy in the severe psoriasis group approaches that in patients with organ transplants (risk ratio, 2.12; 95% CI, 1.80-2.50). Most of these cancers were nonmelanoma skin cancers and lymphoproliferative malignancies. Those with less severe psoriasis were only slightly more likely to develop a new malignancy than those with hypertension (risk ratio, 1.13; 95% CI, 1.03-1.25). CONCLUSIONS: Patients with psoriasis are at an increased risk of developing a malignancy compared with patients with hypertension. The increased risk is greatest for those with severe disease (ie, patients with psoriasis treated with systemic agents) and minimal (if an increased risk at all) for those with less severe disease compared with those in the hypertension group. The increased risk is mainly for lymphoproliferative cancers and nonmelanoma skin cancers. PMID- 11405771 TI - Anti-tumor necrosis factor alpha therapy in psoriatic arthritis and psoriasis. PMID- 11405772 TI - Can cognitive-behavioral therapy help patients with vitiligo? PMID- 11405773 TI - A population-based estimate of the prevalence of postherpetic neuralgia after herpes zoster. PMID- 11405774 TI - Lyme disease has a favorable prognosis. PMID- 11405775 TI - On beauty: evolution, psychosocial considerations, and surgical enhancement. PMID- 11405776 TI - Advancing the science and art of teaching dermatology. Abstracts presented at the 10th Winter Meeting of the Dermatology Teachers Exchange Group, March 12, 2000, San Francisco, Calif. PMID- 11405777 TI - Autologous fat transplantation. PMID- 11405778 TI - Verrucoid lesions of the right hand and wrist. PMID- 11405779 TI - A crusted lip nodule in a 50-year-old man. PMID- 11405780 TI - Intertriginous bullae in a 53-year-old man. PMID- 11405781 TI - A papillary lesion on the foreskin. PMID- 11405782 TI - Delayed EPPER syndrome. PMID- 11405783 TI - Neonatal and infantile erythrodermas. PMID- 11405784 TI - Multiple interpretations of cancer risks from body mole counts in preventive care. PMID- 11405785 TI - Arsenic therapy. PMID- 11405786 TI - Use of the 308-nm excimer laser for postresurfacing leukoderma. PMID- 11405787 TI - The historical and geomedical immunogenetics of pemphigus among the descendants of Sephardic Jews in New Mexico. PMID- 11405788 TI - Malignant melanomas: search for human papillomaviruses. PMID- 11405789 TI - Successful treatment of an intractable case of hereditary basal cell carcinoma syndrome with paclitaxel. PMID- 11405790 TI - Benign neonatal hemangiomatosis with mucosal involvement. PMID- 11405799 TI - Titin, thymoma, and myasthenia gravis. PMID- 11405800 TI - The challenge of respiratory dysfunction in Guillain-Barre syndrome. PMID- 11405801 TI - Plasticity, hippocampal place cells, and cognitive maps. AB - Memory of even the briefest event can last a lifetime. Thus, learning and memory require neuronal mechanisms that allow rapid, yet persistent, changes to brain circuits. Hippocampal neuropsychology, synaptic and cellular electrophysiology, pharmacology, and molecular genetics converge and begin to reveal these mechanisms. Lesions of the hippocampus profoundly impair memory for recent events in humans and rodents. Circuits within the hippocampus are remarkably plastic, and this plasticity is mediated in part through changes in synaptic strength and revealed by long-term potentiation (LTP) and long-term depression (LTD). N-methyl D-aspartate (NMDA) receptors, a subtype of glutamate receptor, are crucial for inducing these plastic changes, and blocking these receptors reduces plasticity and impairs learning in tasks that require the hippocampus. Molecular genetic alterations that disrupt signaling mechanisms downstream of the NMDA receptor also prevent LTP induction and impair hippocampus-dependent learning. N-methyl D aspartate receptor mechanisms have also been linked to information coding by hippocampal neurons. Hippocampal cells fire selectively in specific and restricted locations (place fields) as rodents move through open environments. Place fields form within minutes and persist for months. N-methyl D-aspartate receptor antagonists prevent the establishment of stable place fields. The same molecular genetic manipulations that interfere with hippocampal NMDA receptor function, prevent LTP induction, and impair spatial learning also disrupt the formation of stable hippocampal place fields. Finally, learning has been improved in mice with genetically modified NMDA receptors that enhance LTP induction. Thus, hippocampal cells "learn" to encode the salient features of experience through NMDA receptor-dependent synaptic plasticity mechanisms, and this rapid and persistent neuronal encoding is a crucial step toward the formation of long term memory. Disruption of these plasticity mechanisms may underlie age-related memory deficits. PMID- 11405802 TI - Anti-titin antibodies in myasthenia gravis: tight association with thymoma and heterogeneity of nonthymoma patients. AB - BACKGROUND: Titin is the major autoantigen recognized by anti-striated muscle antibodies, which are characteristic of generalized myasthenia gravis (MG). OBJECTIVE: To seek a correlation between anti-titin antibodies and other features of MG patients, including histopathology, age at diagnosis, anti-acetylcholine receptor (anti-AChR), autoantibody titers, and clinical severity. METHODS: A novel, highly specific radioligand assay was performed on a large group of 398 patients with generalized MG. RESULTS: Among thymectomized patients, anti-titin antibodies were present in most patients with thymoma (56/70 [80%]), contrasting with only a minority of patients with thymus atrophy or hyperplasia (17/165 [10%]). They were also present in 64 (41%) of 155 nonthymectomized patients who had a radiologically normal thymus. In these patients and in those who had a histologically normal thymus, anti-titin antibodies were associated with a later age at onset of disease and with intermediate titers of anti-AChR antibodies. After controlling for these 2 variables, disease severity was not significantly influenced by anti-titin antibodies. CONCLUSIONS: Anti-titin antibodies are a sensitive marker of thymoma associated with MG in patients 60 years and younger, justifying the insistent search for a thymoma in MG patients of this age group who have these antibodies. In nonthymoma patients, anti-titin antibodies represent an interesting marker complementary to the anti-AChR antibody titer, identifying a restricted subset of patients. These clinical correlations should prompt further studies to examine the mechanisms leading to the production of anti-titin antibodies. PMID- 11405803 TI - Anticipating mechanical ventilation in Guillain-Barre syndrome. AB - CONTEXT: The combination of multiple clinical factors culminates in neuromuscular respiratory failure in up to 30% of the patients with Guillain-Barre syndrome (GBS). Although guidelines exist as to when to proceed with intubation, early indicators of subsequent progression to respiratory failure have not been established. OBJECTIVES: To identify clinical and respiratory features associated with progression to respiratory failure and to examine patterns of respiratory decline in patients with severe GBS. DESIGN: Retrospective survey. SETTING: Tertiary care hospital. PATIENTS: One hundred fourteen consecutive patients with severe GBS admitted to the intensive care unit between January 1, 1976, and December 31, 1996. MAIN OUTCOME MEASURES: Early markers of impending respiratory failure, requirement for mechanical ventilation, and patterns of respiratory decline. METHODS: The clinical and electrophysiologic features of 60 patients receiving mechanical ventilation were compared with 54 patients with severe GBS who did not receive mechanical ventilation. Daily preventilation maximal inspiratory and maximal expiratory respiratory pressures and vital capacity were analyzed. Multivariate predictors of the necessity for mechanical ventilation were assessed using logistic regression analysis. RESULTS: Progression to mechanical ventilation was highly likely to occur in those patients with rapid disease progression, bulbar dysfunction, bilateral facial weakness, or dysautonomia. Factors associated with progression to respiratory failure included vital capacity of less than 20 mL/kg, maximal inspiratory pressure less than 30 cm H(2)O, maximal expiratory pressure less than 40 cm H(2)O or a reduction of more than 30% in vital capacity, maximal inspiratory pressure, or maximal expiratory pressure. No clinical features predicted the pattern of respiratory decline; however, serial measurements of pulmonary function tests allowed detection of those at risk for respiratory failure. CONCLUSIONS: While inherently unpredictable, the course of patients with severe GBS can, to some extent, be predicted on the basis of clinical information and simple bedside tests of respiratory function. These data may be used in the decisions regarding admission to the intensive care unit and preparation for elective intubation. PMID- 11405804 TI - Neurologic findings in Machado-Joseph disease: relation with disease duration, subtypes, and (CAG)n. AB - CONTEXT: Machado-Joseph disease (MJD), an autosomal dominant spinocerebellar degeneration caused by an expanded CAG repeat on chromosome 14q32.1, is a heterogeneous disorder for clinical manifestations. The reasons for the wide range of neurologic findings in this disease are poorly understood. OBJECTIVE: To explain part of this heterogeneity through the association of the neurologic findings with sex, disease duration, age of onset, clinical type, and size of CAG repeat expansion. DESIGN: A case-control study. SETTING: Ambulatory care. PATIENTS: A consecutive sample of 62 patients with MJD. MAIN OUTCOME MEASURE: Neurologic signs. RESULTS: A direct relationship was found between the disease duration and severity of gait and limb ataxia, dysarthria, dysphagia, fasciculations, pyramidal syndrome, and ophthalmoplegia (P<.02). The most severe forms of nuclear ophthalmoplegia were associated with type 1 MJD, whereas those of supranuclear ophthalmoplegia were associated with type 3 MJD (P<.001). It was also found that higher mean (CAG)(n) lengths were associated with worse degrees of the pyramidal syndrome and dystonia (P<.001). The presence and severity of nystagmus, eyelid retraction, rigidity and/or bradykinesia, and optic atrophy were not clearly associated with any of the predictive variables under study. CONCLUSIONS: Disease duration can explain part of the heterogeneity of ataxia, dysarthria, dysphagia, fasciculations, pyramidal syndrome, and ophthalmoplegia, in MJD. Type 1 MJD was positively associated with nuclear ophthalmoplegia; type 3 MJD was positively associated with supranuclear ophthalmoplegia. Higher mean CAG lengths were found to correlate with the pyramidal syndrome and dystonia. Nystagmus, eyelid retraction, rigidity and/or bradykinesia, and optic atrophy were hardly attributable to any known reason or variable. PMID- 11405805 TI - Response to levodopa treatment in dopa-responsive dystonia. AB - BACKGROUND: Dopa-responsive dystonia (DRD) is similar to Parkinson disease in that both disorders have impaired dopamine synthesis and respond to levodopa treatment. Dopa-responsive dystonia differs in that dopamine storage is intact in contrast to Parkinson disease in which it is markedly reduced. OBJECTIVE: To examine the short- and long-duration responses to levodopa dosing in subjects with DRD. METHODS: The response to brief infusions of levodopa was examined in 4 subjects with DRD and the effects of withdrawal of levodopa for 3 to 7 days studied in the 3 subjects receiving long-term levodopa therapy. Motor function was measured with tapping speed, Unified Parkinson's Disease Rating Scale motor score, and global dystonia score. RESULTS: The short-duration response to levodopa dosing seems to develop more slowly and persists longer in subjects with DRD than in subjects with Parkinson disease. Withdrawal of levodopa leads to a gradual decline in tapping speed and reemergence of dystonia over several days, similar to the rate of decay of motor function in Parkinson disease. The short- and long-duration responses were not clearly differentiated in DRD. CONCLUSIONS: This pilot study suggests that retained dopamine storage in DRD may prolong the short-duration response and blur the distinction of the short- and long-duration responses. The decline in motor function in DRD on withdrawal of long-term levodopa therapy resembles that in Parkinson disease, suggesting that a long duration response, if it exists in DRD, is unrelated to dopamine storage. PMID- 11405806 TI - Early electrodiagnostic findings in Guillain-Barre syndrome. AB - CONTEXT: Guillain-Barre syndrome (GBS) is the foremost cause of acute, generalized, peripheral neuropathic weakness. Although nerve conduction studies are a diagnostic aid, the characteristic electrical changes may not evolve for several weeks. Early diagnosis of GBS is important, however, because early treatment has been shown to improve outcome. OBJECTIVES: To describe the electrodiagnostic abnormalities detectable in the first week of GBS, to determine if there are early patterns suggestive of GBS, and to identify the percentage of patients whose condition can be diagnosed with reasonable certainty in the first week. DESIGN AND SETTING: We retrospectively reviewed the medical records of all patients admitted to the Cleveland Clinic Foundation, Cleveland, Ohio, having the discharge diagnosis GBS during the past 16 years. Patients who underwent nerve conduction studies within 7 days of muscle weakness were selected for this study. RESULTS: The H reflex was absent in 30 (97%) of 31 patients. Nineteen patients (61%) had low-amplitude or absent sensory nerve action potential (SNAP) in the upper extremity. Fifteen patients (48%) overall, including 21 (67%) of the 31 patients, including 14 (67%) of the 21 patients younger than 60 years, had an abnormal upper extremity SNAP combined with a normal sural SNAP. Other findings included an abnormal F wave (25 patients [84%]), reduced compound muscle action potential amplitude (22 patients [71%]), prolonged distal latency (20 patients [65%]), temporal dispersion (18 patients [58%]), slowed motor conduction velocity (16 patients [52%]), and motor conduction block (4 patients [13%]). Definite diagnosis was possible in 17 patients (55%), but not commonly until the fifth day. CONCLUSIONS: The H reflex is the most sensitive test for early GBS. Upper extremity SNAPs are also frequently abnormal in early GBS. Absent H response, abnormal F wave, and abnormal upper extremity SNAP combined with a normal sural SNAP are characteristic of early GBS. If multiple nerves are tested, definite diagnosis is possible in half the patients, but not until the fifth day after the onset of symptoms. PMID- 11405807 TI - Cellular distribution of torsin A and torsin B in normal human brain. AB - BACKGROUND: Early-onset torsion dystonia is a hyperkinetic movement disorder caused by a deletion of 1 glutamic acid residue in torsin A protein, a novel member of the AAA family of adenosine triphosphatases. No mutation has been found so far in the closely related torsin B protein. Little is known about the molecular basis of the disease, and the cellular functions of torsin proteins remain to be investigated. OBJECTIVE: To study the regional, cellular, and subcellular distribution of the torsin A and torsin B proteins. METHODS: Expression of torsin proteins in the central nervous system was analyzed by Western blot analysis and immunohistochemistry in human postmortem brain tissues. RESULTS: We generated polyclonal antipeptide antibodies directed against human torsin A and torsin B proteins. In Western blot analysis of normal human brain homogenates, the antibodies specifically recognized 38-kd endogenous torsin A and 62-kd endogenous torsin B. Absorption controls showed that labeling was blocked by cognate peptide used for immunization. Immunolocalization studies revealed that torsin A and torsin B were widely expressed throughout the human central nervous system. Both proteins displayed cytoplasmic distribution, although torsin B localization in some neurons was perinuclear. Strong labeling of neuronal processes was detected for both proteins. CONCLUSIONS: Torsin A and torsin B have similar distribution in the central nervous system, although their subcellular localization is not identical. Strong expression in neuronal processes points to a potential role for torsin proteins in synaptic functioning. PMID- 11405808 TI - [18F]FDG-PET reveals temporal hypometabolism in patients with temporal lobe epilepsy even when quantitative MRI and histopathological analysis show only mild hippocampal damage. AB - BACKGROUND: The relationship between reduced glucose metabolism in positron emission tomography with fludeoxyglucose F 18 ([(18)F]FDG-PET) and hippocampal damage (HD) in patients with temporal lobe epilepsy is still unclear. OBJECTIVE: To determine whether the presence and severity of HD verified by quantitative magnetic resonance imaging (QMRI) and histopathological analysis affect the degree of hypometabolism. PATIENTS AND METHODS: Sixteen patients with drug resistant temporal lobe epilepsy underwent [(18)F]FDG-PET and QMRI (hippocampal volumetry and T2 relaxometry) before surgery. Histopathological analysis of the hippocampus included measurements of neuronal loss, proliferation of glial cells, and mossy fiber sprouting. The asymmetry in glucose metabolism described the degree of hypometabolism. RESULTS: Temporal hypometabolism was not related to severity of HD as measured by QMRI or histopathological analysis. The degree of hypometabolism did not differ in patients with mild, moderate, or severe HD. In addition, [(18)F]FDG-PET revealed significant temporal hypometabolism even though hippocampal QMRI findings were normal or showed only mild HD. Thus, glucose consumption was reduced over and above the histopathological changes. CONCLUSIONS: [(18)F]FDG-PET is sensitive for localizing the epileptogenic region in patients with temporal lobe epilepsy. However, it is insensitive to reflect the severity of HD. PMID- 11405809 TI - Oral almotriptan vs. oral sumatriptan in the abortive treatment of migraine: a double-blind, randomized, parallel-group, optimum-dose comparison. AB - BACKGROUND: Almotriptan malate is a novel, selective serotonin(1B/D) agonist, or triptan, developed for the abortive treatment of migraine. In double-blind, placebo-controlled studies, it has been shown to be effective, well tolerated, and safe. OBJECTIVE: To compare the efficacy, tolerability, and safety of almotriptan with that of the "standard triptan," sumatriptan succinate. The power calculation of the study was based on 24-hour headache recurrence, an efficacy measure in the abortive treatment of migraine, and on the occurrence of adverse events. SUBJECTS AND METHODS: Subjects, aged between18 and 65 years, with migraine with or without aura but otherwise healthy, were randomized to take orally either almotriptan malate, 12.5 mg, or sumatriptan succinate, 50 mg. The medications were provided in identical-looking capsules to ensure blinding and were taken for the treatment of moderate or severe headache. Efficacy was determined in terms of (1) headache relief-a decrease in pain intensity to mild or no pain; (2) headache freedom-a decrease to no pain; (3) use of rescue medications, allowed after 2 hours; and (4) headache recurrence-moderate or severe pain returning within 24 hours after headache relief at 2 hours. Adverse events were collected for 96 hours after treatment and for safety evaluation, vital signs, blood tests, and electrocardiograms were performed at the screening and exit visits. RESULTS: Seventy-five investigators enrolled 1255 subjects of whom 1173 were treated (591 with almotriptan and 582 with sumatriptan). At 2 hours, almotriptan treatment provided headache relief in 58.0% of the subjects and sumatriptan treatment in 57.3%; headache freedom was provided by the medications in 17.9% and 24.6%, respectively (P =.005). Rescue medications were taken by 36.7% of the subjects in the almotriptan-treated group and by 33.2% in the sumatriptan-treated group; headaches returned to moderate or severe intensity in 27.4% and 24.0%, respectively. Treatment-emergent adverse events occurred in 15.2% of the subjects in the almotriptan-treated group and in 19.4% in the sumatriptan-treated group (P =.06); treatment-related adverse events occurred in 9.1% and 15.5% of the subjects, respectively (P =.001), including chest pain, which occurred in 0.3% and 2.2%, respectively (P =.004). CONCLUSIONS: Almotriptan and sumatriptan are similarly effective in the abortive treatment of moderate or severe migraine headache; they are also similarly well tolerated and safe. PMID- 11405810 TI - Autopsy-confirmed familial early-onset Alzheimer disease caused by the l153V presenilin 1 mutation. AB - BACKGROUND: Three affected individuals are described from a small English kindred with early-onset autosomal dominant familial Alzheimer disease (FAD) caused by a leucine-to-valine change at codon 153 (L153V) of the presenilin 1 (PSEN1) gene. METHODS: Clinical information on the pedigree was collected directly from family members and from hospital records. Samples of DNA were screened by means of direct sequencing of all coding exons of PSEN1. One patient underwent neuropathological examination. RESULTS: Mean age at onset of symptoms was 35.3 years (95% confidence interval [CI], 34.6-36.0 years); at death, 44.0 years (95% CI, 39.1-48.9 years). Mean duration of illness was 8.3 years (95% CI, 4.7-11.9 years). Myoclonus was a late feature in 1 patient; seizures were not reported in any subjects. Spastic paraparesis and extrapyramidal signs were absent. The neuropsychometric profile of 1 patient showed relatively preserved naming skills in the setting of global cognitive deficits. Results of neuropathological examination demonstrated the signature lesions of Alzheimer disease and the presence of occasional cortical Lewy bodies. CONCLUSIONS: The PSEN1 L153V mutation lies in the main mutation cluster of PSEN1 in the second transmembrane domain. It causes early-onset FAD with clinical features similar to those of other reported FAD pedigrees. PMID- 11405811 TI - Use of the multiple sclerosis functional composite as an outcome measure in a phase 3 clinical trial. AB - BACKGROUND: The Multiple Sclerosis Functional Composite (MSFC) is a multidimensional clinical outcome measure that includes quantitative tests of leg function/ambulation (Timed 25-Foot Walk), arm function (9-Hole Peg Test), and cognitive function (Paced Auditory Serial Addition Test). The MSFC is the primary outcome measure in the ongoing multinational phase 3 trial of interferon beta-1a (Avonex) in patients with secondary progressive MS. OBJECTIVE: To assess the practice effects, reliability, and validity of the MSFC clinical outcome measure. DESIGN: Examining technicians underwent formal training using standardized materials. The MSFC was performed according to a standardized protocol. The 436 patients enrolled in the International Multiple Sclerosis Secondary Progressive Avonex Controlled Trial underwent 3 prebaseline MSFC testing sessions before randomization. RESULTS: Practice effects were evident initially for the MSFC but stabilized by the fourth administration. The Paced Auditory Serial Addition Test demonstrated the most prominent practice effects. The reliability of the MSFC was excellent, with an intraclass correlation coefficient for session 3 (final prebaseline session) vs session 4 (baseline) of 0.90. The MSFC at baseline correlated moderately strongly with the Kurtzke Expanded Disability Status Scale. Among the MSFC components, the Timed 25-Foot Walk correlated most closely. Correlations among the 3 MSFC components were weak, suggesting they assess distinct aspects of neurologic function in patients with MS. CONCLUSIONS: The MSFC demonstrated excellent intrarater reliability in this multinational phase 3 trial. Three prebaseline testing sessions were sufficient to compensate for practice effects. The pattern of correlations among the MSFC, its components, and the Kurtzke Expanded Disability Status Scale supported the validity of the MSFC. PMID- 11405812 TI - Mutation analysis and the correlation between genotype and phenotype of Arg778Leu mutation in chinese patients with Wilson disease. AB - BACKGROUND: The defective gene (ATP7B) that causes Wilson disease (WD) codes for a putative copper-transporting P-type adenosine triphosphatase. After cloning of ATP7B, the spectrum of mutations and their clinical consequences have been investigated in patients with WD in different ethnic populations. However, the spectrum of mutations and the correlation of genotype-phenotype in the Chinese population have not been extensively studied. OBJECTIVE: To investigate the characterization of mutations of ATP7B and the correlation between genotype and phenotype in the Chinese population. METHODS: We studied 60 unrelated healthy Chinese and 65 unrelated Chinese families, including 84 patients with WD and 126 parents. Genomic DNA was prepared from peripheral blood leukocytes using a salt precipitation method. Polymerase chain reaction single-strand conformation polymorphism and subsequent direct sequencing were used to identify the mutations and polymorphisms of ATP7B. Statistical analysis was performed using t test or chi(2) test. RESULTS: We identified 18 mutations (7 novel) and 11 polymorphisms (3 novel). The novel mutations are -36C-->T, Trp650ter, Gln914ter, 2810delT, Thr935Met, Arg1041Pro, and Glu1173Lys. The novel polymorphisms are 1168A-->G (Ile390Val), 2785A-->G (Ile929Val), and 3316G-->A (Val1106Ile). Two mutations, Arg778Leu and Thr935Met, are relatively frequent, representing 37.7% and 10.0% of patients, respectively. To our knowledge, we are the first to report the correlation between the genotype and phenotype of Arg778Leu. The result shows that Arg778Leu homozygotes are associated with the early onset of WD with hepatic presentation. CONCLUSIONS: The Arg778Leu and Thr935Met mutations are hot spots in the Chinese population. The features of mutations of ATP7B differ between the Chinese and Western ethnic populations. The Arg778Leu mutation has severe effects on the function of ATP7B. These findings are valuable for developing a fast and effective method to diagnose the presence of the WD gene. PMID- 11405813 TI - Attention and fluctuating attention in patients with dementia with Lewy bodies and Alzheimer disease. AB - BACKGROUND: Attentional deficits are described in the consensus clinical criteria for the operationalized diagnosis of dementia with Lewy bodies (DLB) as characteristic of the condition. In addition, preliminary studies have indicated that both attentional impairments and fluctuation of attentional impairments are more marked in patients with DLB than in patients with Alzheimer disease (AD), although neuropsychological function has not previously been examined in a large prospective cohort with confirmed diagnostic accuracy against postmortem diagnosis. METHODS: A detailed evaluation of attention and fluctuating attention was undertaken in 155 patients with dementia (85 with DLB and 80 with AD) from a representative hospital dementia case register and 35 elderly controls using the Cognitive Drug Research Computerized Assessment System for Dementia Patients computerized neuropsychological battery. Operationalized clinical diagnosis was made using the consensus criteria for DLB and the National Institute of Neurological and Communicative Disorders and Stroke-Alzheimer's Disease and Related Disorders Association criteria for AD. High levels of sensitivity and specificity have been achieved for the first 50 cases undergoing postmortem examination. RESULTS: The groups were well matched for severity of cognitive impairments, but the AD patients were older (mean age, 80 vs 78 years) and more likely to be female (55% vs 40%). Patients with DLB were significantly more impaired than patients with AD on all measures of attention and fluctuating attention (for all comparisons, t > or = 2.5, P<.001), and patients from both dementia groups were significantly more impaired than elderly controls for all comparisons other than cognitive reaction time, which was significantly more impaired in DLB patients than controls but was comparable in controls and AD patients. There were, however, significant associations between the severity of cognitive impairment and the severity of both attentional deficits and fluctuations in attention. CONCLUSIONS: This large prospective study confirms that slowing of cognitive processing, attention, and fluctuations of attention are significantly more pronounced in DLB and AD patients, although fluctuating attention is common in patients with moderate-to-severe AD. Deficits of cognitive reaction time appear to be specific to DLB, except in severe dementia. A detailed evaluation of attentional performance could make an important contribution to differential diagnosis, although the results need to be interpreted within the context of the overall severity of cognitive deficits. PMID- 11405814 TI - An apparently sporadic case with parkin gene mutation in a Korean woman. AB - OBJECTIVE: To report the clinical features and results of iodine I 123-2beta carbomethoxy-3beta-(4-iodophenyl)-tropane (CIT) single photon emission computed tomography and molecular genetic analysis in a Korean woman with juvenile Parkinson disease with deletion in exon 4 of the parkin gene. DESIGN: Case report with molecular genetic analysis. PATIENT AND RESULTS: The patient had bradykinesia, postural imbalance, and postural tremor since the age of 12 years. She developed wearing off early in the disease course. The [(123)I]-2beta carbomethoxy-3beta-(4-iodophenyl)-tropane single photon emission computed tomography showed severe reduction of specific striatal CIT binding, comparable to that of Parkinson disease. The polymerase chain reaction products from the parkin gene showed homozygous exon 4 deletion. CONCLUSION: In this sporadic juvenile Parkinson disease case, severe nigrostriatal dopaminergic damage and homozygous exon 4 deletion in the parkin gene were demonstrated. PMID- 11405815 TI - Diffusion-weighted magnetic resonance imaging in nonconvulsive status epilepticus. AB - BACKGROUND: In human and experimental models, diffusion-weighted magnetic resonance imaging (DWI) findings in status epilepticus (SE) have been reported to show that apparent diffusion coefficients are reduced during the initial phase and normalized or increased in the later phase of prolonged SE. This effect is caused by cytotoxic edema induced by excitotoxicity. In humans, only focal DWI abnormalities have been reported in partial SE. OBJECTIVES: To report and discuss the DWI findings suggesting diffuse neuronal injury in a patient with nonconvulsive SE. DESIGN AND METHODS: A 56-year-old man was admitted because of changing levels of consciousness over 3 days. On admission he was comatose. He had nystagmoid eye movement, forced eye blinking, and oroalimentary automatism. The results of a search for possible infectious and metabolic etiologies were negative, and electroencephalographic findings showed continuous, semirhythmic, bifrontal sharp waves of 2 Hz. Phenytoin and midazolam hydrochloride were infused to alleviate the seizure activities. He underwent DWI initially (3 days after the onset of seizure) and at the 5-month follow-up. SETTING: The neurology department of a tertiary referral center. RESULTS: During SE, DWI findings showed marked, diffuse gyriform cortical hyperintensity throughout the brain. The apparent diffusion coefficient decreased in the corresponding areas, especially in the occipital lobes. Findings from T2-weighted magnetic resonance imaging showed the intense cortical hyperintensity with gyral swelling and no involvement of brainstem, basal ganglia, thalamus, and white matter. The follow-up DWI findings showed marked atrophy and hypointensity in the corresponding regions. The apparent diffusion coefficient increased in the corresponding regions. CONCLUSIONS: Diffusion-weighted imaging in our patient indicated that the magnetic resonance imaging abnormalities of the affected cortex were due to cytotoxic edema caused by neuronal excitotoxicity during prolonged SE. Diffusion weighted imaging can be used in the localization of seizure focus for predicting the prognosis of the affected tissue and for researching the basic pathophysiology of SE. PMID- 11405816 TI - A family with X-linked dystonia-deafness syndrome with a novel mutation of the DDP gene. AB - BACKGROUND: X-linked dystonia-deafness syndrome (DDS) is characterized by early onset deafness followed by progressive dystonia in adulthood. Only 4 families with the syndrome have been reported, and all were white. OBJECTIVE: To describe the first nonwhite family with X-linked DDS, involving 5 affected males in 4 generations. RESULTS: Clinical features of the family members, who were Japanese, were mostly consistent with reports of DDS in whites except for a lack of visual disturbances. Whereas microdeletions in the deafness-dystonia peptide (DDP) gene were found in 2 white DDS families, our patients showed a novel mutation (arg80ter) in exon 2 of the DDP gene. CONCLUSION: The existence of a DDS family of Japanese origin with a new kind of mutation in the DDP gene provides additional evidence that the DDP gene is a causative gene for X-linked DDS. PMID- 11405817 TI - Macular star in neuroretinitis. PMID- 11405818 TI - Clinical brainstem death with preserved electroencephalographic activity and visual evoked response. PMID- 11405819 TI - Emergence of temporal lobe surgery for epilepsy. PMID- 11405820 TI - Charcot-Marie-tooth disease. PMID- 11405821 TI - Neurology was there in the 1930s. PMID- 11405822 TI - Coexistence of 2 different type I repeats. PMID- 11405823 TI - Statin-Alzheimer disease association not yet proven. PMID- 11405825 TI - Statin therapy and the prevention of dementia. PMID- 11405827 TI - Neurology was there: 1929. PMID- 11405828 TI - T2-hyperintense lateral rim and hypointense putamen are typical but not exclusive of multiple system atrophy. PMID- 11405830 TI - Use of tissue plasminogen activator to revive blebs following intraocular surgery. AB - BACKGROUND: Tissue plasminogen activator (tPA) is a serine protease with thrombolytic-fibrinolytic activity. In the anterior segment of the eye, it has been used to lyse blood and fibrin clots immediately following trabeculectomy. OBJECTIVE: To review our experience using tPA to revive previously functional but newly failing blebs following secondary surgical procedures (cataract extraction or penetrating keratoplasty). METHODS: A retrospective medical record review of all eyes receiving tPA to revive a failing bleb after a later anterior segment surgery was performed. Blebs were functional for at least 9 months before tPA, 12.5 microg, was injected into the anterior chamber. Eyes had longer than 6 months of follow-up after tPA use. RESULTS: Six eyes that had undergone phacoemulsification and 1 that had undergone penetrating keratoplasty were identified. Before the secondary surgery, blebs had been in place for 20.4 +/- 9.3 months, with an intraocular pressure of 10.7 +/- 3.6 mm Hg. New bleb failure in these eyes was observed following phacoemulsification or penetrating keratoplasty between postoperative days 4 and 14, with the intraocular pressure increasing to 27.7 +/- 9.5 mm Hg. One day after tPA use, the intraocular pressure had decreased to 11.0 +/- 3.7 mm Hg (P =.002). In 5 of the 7 patients, the bleb height improved following tPA use. The final intraocular pressure was 10.7 +/- 3.6 mm Hg at a follow-up of 13.4 +/- 6.4 months. (All data are given as mean +/- SD.) CONCLUSION: Tissue plasminogen activator can be a useful adjunct for reviving newly failing blebs after other anterior segment surgery. PMID- 11405831 TI - Clinical factors associated with progression of glaucomatous optic disc damage in treated patients. AB - BACKGROUND: Reducing intraocular pressure (IOP) in glaucomatous eyes does not always prevent disease progression. OBJECTIVE: To determine the clinical factors associated with progressive optic disc damage in glaucomatous eyes receiving treatment to reduce IOP. METHODS: Baseline and follow-up optic disc photographs as well as demographic and clinical data were retrospectively studied in 186 eyes of 93 patients with primary open-angle glaucoma, and in 138 eyes of 69 patients with normal-pressure glaucoma. The patients with primary open-angle glaucoma were included in the study only if their treated IOPs during a follow-up period of 5 years were less than 21 mm Hg. The patients with normal-pressure glaucoma were included only if their IOPs were reduced by at least 20% during the follow-up period. The association of progressive optic disc damage with patient- and eye specific characteristics was examined using multivariate analysis. RESULTS: During the 5-year study period, 141 (43.5%) of the 324 eyes exhibited progressive optic disc damage defined by at least a 5% decrease in the neural rim area-to disc area ratio. Using multivariate analysis, the following were found to be strongly associated with progressive neural rim damage: a baseline smaller neural rim area-disc area ratio (P<.001); a baseline larger zone beta area-disc area ratio (P =.04); a baseline larger parapapillary atrophy length-disc circumference ratio (P =.05); a diagnosis of normal-pressure glaucoma (P =.01); and combined medical and surgical treatment prior to the study period (P =.01). CONCLUSIONS: Clinical factors other than IOP may be important indicators of subsequent progression of glaucomatous optic disc damage. Our findings suggest that eyes with advanced glaucomatous optic disc damage and normal-pressure glaucoma are more likely to progress despite receiving treatment to reduce IOP. PMID- 11405832 TI - Clinical investigation of a true color scanning laser ophthalmoscope. AB - OBJECTIVE: To show true color scanning laser ophthalmoscope (SLO) images produced by simultaneously imaging the retina with red, green, and blue lasers. METHODS: Low-power red, green, and blue lasers were combined using fiber optics. By rapidly pulsing the lasers, each point on the fundus is illuminated by the 3 colors in quick succession, with the total power level being similar to that from a single laser. The reflected light is then decoded to extract the red, green, and blue color information and the true color fundus image is displayed live on a computer monitor. RESULTS: Comparison was made between the color SLO images from 5 patients and their digitized fundus photographs. The background fundus and retinal vasculature showed a similar appearance. The SLO gave better quality information in patients with ocular histoplasmosis, macular dystrophy, and optic disc drusen. By operating the color SLO in the indirect mode, macular edema could be clearly seen as lines and ridges surrounding the fovea. CONCLUSION: The color SLO offers all the advantages of the present commercially available monochromatic device, with the added advantage of true color representation of the fundus without increasing either imaging time or the level of exposure. PMID- 11405833 TI - Laser burn intensity and the risk for choroidal neovascularization in the CNVPT Fellow Eye Study. AB - OBJECTIVE: To explore the relationship between laser burn intensity and the subsequent risk for development of choroidal neovascularization (CNV) in eyes assigned to the treatment group of the Fellow Eye Study (FES) of the Choroidal Neovascularization Prevention Trial (CNVPT), using computerized methods for laser burn quantitation, and to examine the association between laser burn intensity and (1) drusen reduction and (2) visual acuity. METHODS: Color fundus images before and immediately after laser treatment in the CNVPT FES were available for 53 of 59 eyes. Prelaser and postlaser treatment images were analyzed using custom developed computer software, allowing for laser burn identification and quantitation. As measures of laser burn intensity, we derived integrated burn rating (IBR) (the integral of the normalized intensity difference divided by the burn pixels), and the maximum burn intensity (MAX). We identified CNV using fluorescein angiography. A Cox proportional hazards model was fit to the time to development of CNV. Baseline and 6-month color photographs were used to determine reduction in drusen. Visual acuity was measured using a standardized protocol. RESULTS: The IBR and MAX spanned 4.5 logarithm units. After adjusting for smoking history and predominant drusen size, the risk ratio for CNV per logarithm unit of increasing laser burn intensity for each measure was 2.0 (P =.05) for MAX and 1.7 (P =.07) for IBR. When patients were divided into high- and low-intensity treatment groups of equal size, the high-intensity group had more drusen reduction (57% vs 32%; P =.14). There was no effect of laser intensity on change in visual acuity at 6 months. CONCLUSION: Higher-intensity prophylactic laser applications appear to be associated with a greater risk for development of CNV and with more extensive drusen reduction. PMID- 11405834 TI - Sclerochoroidal calcification: clinical manifestations and systemic associations. AB - BACKGROUND: Sclerochoroidal calcification is an unusual ocular condition that is believed to be idiopathic in most cases. OBJECTIVES: To describe the clinical manifestations of sclerochoroidal calcification and to investigate its possible systemic associations. METHODS: This noncomparative consecutive case series included patients diagnosed as having sclerochoroidal calcification based on clinical characteristics and diagnostic test findings. We analyzed the demographic, systemic, and ocular features of 27 such patients. Systemic evaluation included tests for calcium-phosphorus metabolism in 19 patients and renal tubular hypokalemic metabolic alkalosis syndromes (Bartter or Gitelman syndrome) in 13. RESULTS: All the patients were asymptomatic older (mean age, 70 years) white individuals, incidentally noted as having a choroidal lesion on routine examination. Among 38 eyes, the main referral diagnoses were choroidal metastasis in 10 eyes (26%), choroidal melanoma in 8 (21%), and choroidal nevus in 4 (11%). Sixteen patients (59%) had unilateral clinical findings, while 11 (41%) had bilateral. The Snellen visual acuity was 20/50 or better in 37 eyes (97%). Cogan scleral plaque was visible anterior to the insertion of horizontal rectus muscles in 10 eyes (26%). Among 77 foci, there were a mean of 2 foci of sclerochoroidal calcification in each eye, 41 yellow (53%), 32 yellow-white (42%), 2 white (3%), and 2 orange (3%), measuring a mean 2.6 mm in diameter and 1.1 mm in thickness. The most common locations were postequatorial in 45 (58%), along the temporal vascular arcades in 30 (39%), and in the superotemporal quadrant in 43 (56%). A-scan and B-scan ultrasonography revealed dense echoes compatible with calcium, with orbital shadowing. All the lesions remained stable in size and configuration during a mean follow-up of 38 months. One patient developed a choroidal neovascular membrane over the area of sclerochoroidal calcification. Investigations for abnormal calcium-phosphorus metabolism in 19 patients revealed primary hyperparathyroidism in 1 patient (5%). Clinical and biochemical evaluation of 13 patients demonstrated hypomagnesemia in 6 (46%). Four patients (31%) met the criteria for the diagnosis of Gitelman syndrome. CONCLUSIONS: Sclerochoroidal calcification usually manifests as multiple discrete yellow placoid lesions in the midperipheral fundus of asymptomatic older white individuals. Although most cases may be idiopathic in nature, some patients may have underlying systemic disorders involving abnormal calcium-phosphorus metabolism or renal tubular hypokalemic metabolic alkalosis syndromes. All patients with sclerochoroidal calcification should be tested for these treatable systemic associations. PMID- 11405835 TI - Visual functioning and general health status in patients with uveitis. AB - OBJECTIVE: To measure the visual functioning and quality of life in patients with uveitis. METHODS: Consecutive adult patients with noninfectious uveitis were enrolled. The Medical Outcomes Study 36-Item Short Form (SF-36) and the National Eye Institute Visual Functioning Questionnaire (NEI VFQ-25) were administered by a trained interviewer. Sociodemographic and clinical data were also collected. RESULTS: Seventy-six patients were enrolled. The overall NEI VFQ-25 score was significantly lower among patients with uveitis than in a normal reference group (P<.001). The SF-36 physical (PCS) and mental (MCS) component summary scores were also significantly lower among patients with uveitis than in the general US population (P<.007). Among patients with uveitis, visual acuity, binocular involvement, intensity of therapy, employment status, and PCS and MCS scores were all significantly associated with overall NEI VFQ-25 scores in multivariable analysis. Medical comorbidity, ocular comorbidity, and NEI VFQ-25 scores were significantly associated with PCS scores. Medical comorbidity and NEI VFQ-25 scores were significantly associated with MCS scores. Regression models including NEI VFQ-25 scores explained an additional 7% of the variance in PCS scores and 16% of MCS scores. Models including both PCS and MCS scores explained an additional 12% of the variance in NEI VFQ-25 scores. CONCLUSIONS: Patients with uveitis reported markedly poorer visual functioning and general health status than normal subjects. Patients with more severe uveitis have poorer visual functioning and general health status than patients with milder disease. Visual functioning and general health status measurement contribute complementary information and should both be performed in patients with uveitis to measure the effect of disease and its therapy on their quality of life. PMID- 11405836 TI - Induction of tyrosinase gene transcription in human iris organ cultures exposed to latanoprost. AB - OBJECTIVE: Topical administration of latanoprost sometimes induces gradual iris darkening. The present study was undertaken to determine if latanoprost can increase transcription of the gene for tyrosinase, an important enzyme in the biosynthesis of melanin. Results from brown, hazel, and blue irides were compared. METHODS: Iris tissue was isolated from 30 pairs of postmortem human donor eyes, and 2 iris segments from each eye were incubated in tissue culture medium supplemented with 200nM latanoprost acid or vehicle for 7 days. Tyrosinase messenger RNA (mRNA) was determined using real-time polymerase chain reaction analysis (TaqMan quantitative polymerase chain reaction). Results for tyrosinase mRNA were normalized according to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA in each sample. RESULTS: Tyrosinase mRNA expression was similar in blue and hazel irides, and ranged from 0.7-fold to 12.6-fold greater than GAPDH expression. In contrast, control brown iris culture tyrosinase expression ranged from 6.4-fold to 265-fold greater than GAPDH expression. Induction of tyrosinase mRNA by latanoprost was below threshold in all the blue iris cultures (n = 8 pairs), present in 1 of 9 hazel iris cultures, and present in 5 of 13 brown iris cultures. Mean induction in the responding hazel iris cultures was 1.40-fold. Mean induction among the responding brown iris cultures was 2.97-fold. CONCLUSIONS: These observations support the view that iris darkening associated with latanoprost treatment reflects induction of tyrosinase expression. Moreover, they suggest that the probability that latanoprost will increase tyrosinase expression is directly related to the magnitude of tyrosinase expression before treatments are initiated. CLINICAL RELEVANCE: The variability of iris darkening with latanoprost may reflect natural variation in the basal transcription of tyrosinase. PMID- 11405837 TI - Endothelial cell hypertrophy induced by vascular endothelial growth factor in the retina: new insights into the pathogenesis of capillary nonperfusion. AB - OBJECTIVE: To investigate the mechanism leading to capillary nonperfusion of the retina in a monkey model of vascular endothelial growth factor A (VEGF)-induced retinopathy in which capillary closure occurs in a late stage after VEGF treatment. METHODS: Two monkeys received 4 intravitreous injections of 0.5 microg of VEGF in one eye and of phosphate-buffered saline in the other eye and were killed at day 9. After perfusion and enucleation, retinal samples were snap frozen for immunohistochemical analysis with the panendothelial cell marker CD31 or were fixed for morphometric analysis at the light and electron microscopic level. RESULTS: At the light microscopic level, all capillaries in the retina of VEGF-injected eyes displayed hypertrophic walls with narrow lumina. In a quantitative analysis of the deep capillary plexus in the inner nuclear layer, VEGF-injected eyes had a significant 5- to 7-fold decrease in total capillary luminal volume. CD31 staining showed that this decrease was not accompanied by a change in the number of capillaries. Electron microscopy revealed that the luminal volume of individual capillaries of the inner nuclear layer of VEGF injected eyes was significantly decreased due to a 2-fold hypertrophy of the endothelial cells. CONCLUSIONS: Luminal narrowing caused by endothelial cell hypertrophy occurs in the deep retinal capillary plexus in VEGF-induced retinopathy in monkeys. This suggests a causal role of endothelial cell hypertrophy in the pathogenesis of VEGF-induced retinal capillary closure. A similar mechanism may operate in retinal conditions in humans associated with ischemia and VEGF overexpression. CLINICAL RELEVANCE: Capillary nonperfusion occurs in diabetic retinopathy and other ischemic diseases associated with overexpression of VEGF. In addition, VEGF-induced endothelial cell hypertrophy may be causative for capillary closure in these diseases. PMID- 11405838 TI - The color difference in orbital fat. AB - OBJECTIVE: To identify and quantify carotenoids found in white and yellow orbital fat. METHODS: Specimens of nasal (white) and preaponeurotic (yellow) orbital fat were obtained from patients during upper eyelid blepharoplasty. Carotenoids and retinoids were extracted and subjected to spectral and high-performance liquid chromatography analyses. RESULTS: The chromophore content of extracts from unsaponified fat, as measured by absorbance at 425 nm per gram of fat, was 2- to 4-fold higher in preaponeurotic fat than in nasal fat. High-performance liquid chromatography analysis from enzymatically digested fat revealed large amounts of lutein, beta-carotene, and retinol and small amounts of other unidentified carotenoids. The amount of beta-carotene and lutein in preaponeurotic fat was approximately 4-fold higher than in nasal fat. CONCLUSIONS: The higher carotenoid content of preaponeurotic fat might cause it to be more yellow than other orbital fat, and lutein and beta-carotene might be selectively absorbed from plasma by preaponeurotic fat. CLINICAL RELEVANCE: The results provide baseline information for studies of the physiological features of orbital fat in normal and diseased conditions. PMID- 11405839 TI - Association of demographic, familial, medical, and ocular factors with intraocular pressure. AB - OBJECTIVE: To describe the distribution and associations of demographic, familial, medical, and ocular factors with intraocular pressure (IOP). METHODS: A cluster stratified random sample of urban and rural residents of Victoria, Australia, aged 40 years and older. Participants completed an interview and underwent a standardized dilated ophthalmic examination including measurement of IOP with an electronic applanation tonometer (Tono-Pen). Glaucoma status (possible, probable, definite) was determined by a consensus panel. The main outcome measure was IOP. RESULTS: The mean age of the 4576 participants was 59 years, 53% were women, 32% were born overseas, and 132 had open-angle glaucoma. Geometric mean (SD) IOP was 14.3 (+/-1.5) mm Hg. The relationship between IOP and nuclear sclerosis, iris color, and family history of glaucoma depended on glaucoma status. In those with glaucoma, family history of glaucoma and country of birth were significantly associated with IOP in multivariate models (model: r(2) = 0.08, P =.01). In the group without glaucoma, place of residence, use of alcohol, iris color, vitamin E intake, and spherical equivalent were associated with IOP (model: r(2) = 0.01, P =.006). CONCLUSION: In participants with glaucoma, genetic factors seem to be stronger predictors of IOP, whereas in those without glaucoma, lifestyle and physiological factors seem to play a greater role. PMID- 11405840 TI - Visual risk factors for crash involvement in older drivers with cataract. AB - BACKGROUND: The Impact of Cataracts on Mobility project has previously demonstrated that older drivers with cataract have an elevated risk of motor vehicle collision. OBJECTIVE: To examine what types of visual impairment serve as a basis of the increased crash risk of older drivers with cataract. METHODS AND DESIGN: A cross-sectional analysis was performed on 274 older drivers with cataract and 103 older drivers free of cataract recruited through 12 eye care clinics for the purposes of the Impact of Cataracts on Mobility project, a prospective study on driving mobility in older adults with cataract. Tests measured visual acuity, contrast sensitivity, and disability glare for each eye separately using habitual distance correction. The dependent variable was involvement in at least 1 state-recorded, at-fault vehicle crash during the 5 years prior to study enrollment. RESULTS: Logistic regression evaluated associations (odds ratios [ORs]) between visual function and crash involvement. Better and worse eye models defined on the basis of visual acuity were developed. Associations between each type of visual function and crash involvement were adjusted for age, sex, driving exposure, cognitive status, and other types of visual function. For both the better and worse eye models, contrast sensitivity was independently associated with crash involvement, whereas visual acuity and disability glare were not. Drivers with a history of crash involvement were 8 times more likely to have a serious contrast sensitivity deficit in the worse eye (defined as a Pelli-Robson score of 1.25 or less) than those who were crash-free (OR = 7.86; 95% confidence interval [CI], 1.55-39.79); this association was weaker for the better eye but still statistically significant (OR = 3.78; 95% CI, 1.15-12.48). Crash-involved drivers were 6 times more likely to have severe contrast sensitivity impairment in both eyes (OR = 5.78; 95% CI, 1.87-17.86) than crash-free drivers. A severe contrast sensitivity deficit in only 1 eye was still significantly associated with crash involvement (OR = 2.70; 95% CI, 1.16-6.51). CONCLUSION: Severe contrast sensitivity impairment due to cataract elevates at fault crash risk among older drivers, even when present in only 1 eye. PMID- 11405841 TI - The wound healing response after laser in situ keratomileusis and photorefractive keratectomy: elusive control of biological variability and effect on custom laser vision correction. AB - Biological diversity in the wound healing response is thought to be a major factor limiting the predictability of the outcome of refractive surgical procedures such as laser in situ keratomileusis and photorefractive keratectomy. Corneal wound healing is critical to the success of topography-linked or wave front-linked excimer laser ablation to optimize visual performance. This is because of the importance of retaining subtle features of custom ablation and the tendency of epithelial hyperplasia and stromal remodeling to obscure these features following either procedure. The corneal wound healing response is exceedingly complex. Keratocyte apoptosis, which occurs in response to epithelial injury, is the earliest observable event in the wound healing cascades and is therefore an excellent target for pharmacological intervention. Alterations of surgical technique can be designed to limit keratocyte apoptosis and the subsequent events in corneal wound healing. Abnormalities of the cascades could contribute to the pathogenesis of corneal diseases. For example, recent data have suggested that perturbation of the keratocyte apoptosis/mitosis balance could underlie the development of keratoconus in a proportion of patients. PMID- 11405842 TI - Risk factors for glaucoma progression: Where does intraocular pressure fit in? PMID- 11405843 TI - Ethics and human fetal retinal pigment epithelium transplantation. PMID- 11405844 TI - Unilateral follicular conjunctivitis with retained hair and pseudomonal infection. PMID- 11405845 TI - Spontaneous incomplete avulsion of juxtafoveal retinal pigment epithelial hamartoma. PMID- 11405846 TI - Traumatic corneal perforation with epithelial ingrowth after laser in situ keratomileusis. PMID- 11405847 TI - Retinal detachment with a dislocated posterior chamber intraocular lens. PMID- 11405848 TI - Serous macular detachments in a patient with IgM paraproteinemia: an optical coherence tomography study. PMID- 11405849 TI - Bilateral retinal hemorrhages in a preterm infant with retinopathy of prematurity immediately following cardiopulmonary resuscitation. PMID- 11405850 TI - Maternal antiphospholipid antibodies and vitreous hemorrhages in the newborn: a case report. PMID- 11405851 TI - Exogenous endophthalmitis caused by amphotericin B-resistant Paecilomyces lilacinus: treatment options and visual outcomes. PMID- 11405852 TI - Retinal nerve fiber layer loss after traumatic optic neuropathy detected by scanning laser polarimetry. PMID- 11405853 TI - Frontoethmoidal mucoceles causing bilateral chorioretinal folds. PMID- 11405854 TI - Temporary retrobulbar catheter for local anesthesia in vitreoretinal surgery. PMID- 11405856 TI - Iris melanomas in children. PMID- 11405858 TI - The beneficial effect of diclofenac sodium in the treatment of filamentary keratitis. PMID- 11405859 TI - Management of large filtering blebs without surgery. PMID- 11405860 TI - Selective inner hair cell loss in premature infants and cochlea pathological patterns from neonatal intensive care unit autopsies. AB - BACKGROUND: Deafness and handicapping sensorineural hearing impairment occur frequently in neonatal intensive care unit survivors for unknown reasons. PATIENTS AND METHODS: Hearing was tested early and repeatedly in neonatal intensive care unit patients with an auditory brainstem response (ABR) screener. The temporal bones of 15 nonsurvivors (30 ears) were fixed promptly (average, 5 hours) after death for histological evaluation. RESULTS: Among these patients, 12 failed the ABR screen bilaterally, 1 passed unilaterally, and 2 passed bilaterally. Cochlear histopathologic conditions that could contribute to hearing loss included bilateral selective outer hair cell loss in 2 patients, bilateral selective inner hair cell loss in 3 (all premature), and a combination of both outer and inner hair cell loss in 2. Other hair cell abnormalities were noted; the 2 infants who had passed the ABR screen demonstrated normal histological features. Neuronal counts were normal. CONCLUSIONS: Auditory brainstem response failure among these neonatal intensive care unit infants who died was extremely common in part owing to an unexpected histological alteration, selective inner hair cell loss among premature newborns, that should be detectable uniquely by the ABR testing method. Additional histological patterns suggest more than one cause for neonatal intensive care unit hearing loss. Hair cell loss patterns seem frequently compatible with in utero damage. PMID- 11405861 TI - High-resolution imaging of the middle ear with optical coherence tomography: a feasibility study. AB - BACKGROUND: Optical coherence tomography (OCT) is a new medical imaging technology that generates cross-sectional images of tissue microstructure with micron-scale resolution. Optical coherence tomography is analogous to ultrasound, measuring the intensity of infrared light rather than acoustical waves. OBJECTIVE: To demonstrate the feasibility of using OCT for ultra-high-resolution imaging of the middle ear via ex vivo imaging studies of human tissue. DESIGN: Images of the tympanic membrane and middle ear were acquired ex vivo, through the ear canal, without perforating the tympanic membrane. MATERIALS: Four excised intact temporal bones and the auditory apparatus were harvested from cadavers and imaged fresh, without previous fixation. RESULTS: The resulting images were compared with the gross sample and verified the ability of OCT to delineate relevant structures, such as the tympanic membrane and its sublayers, and the middle ear ossicles, nerves, and tendons at higher resolutions than possible with standard clinical imaging technologies. CONCLUSION: The ability of OCT to produce high-resolution images of tissue structure, without contact and in real time, as well as its ability to be integrated with endoscopes, suggests that this technology could become a useful modality for the diagnosis and management of a range of clinical middle ear abnormalities. PMID- 11405862 TI - Significance of airborne transmission of methicillin-resistant Staphylococcus aureus in an otolaryngology-head and neck surgery unit. AB - OBJECTIVES: To quantitatively investigate the existence of airborne methicillin resistant Staphylococcus aureus (MRSA) in a hospital environment and to perform phenotyping and genotyping of MRSA isolates to study MRSA epidemiology. DESIGN: Prospective surveillance of patients with MRSA infections or colonizations was performed, as was an observational study of environmental airAirborne samples were taken by an air sampler; samples were obtained from object surfaces by stamping or swabbing. Epidemiological study of MRSA isolates was performed with an antibiotic susceptibility test, coagulase typing, and pulsed-field gel electrophoresis. SETTING: Three single-patient rooms in a 37-bed otolaryngology head and neck surgery unit. PATIENTS: Three patients with squamous cell head and neck cancer were observed to have been colonized or infected with MRSA after surgery. RESULTS: The MRSA samples were collected from the air in single-patient rooms during both a period of rest and when bedsheets were being changed. Isolates of MRSA were detected in all stages (from stage 1 [>7 microm] to stage 6 [0.65-1.1 microm]). About 20% of the MRSA particles were within a respirable range of less than 4 microm. Methicillin-resistant S aureus was also isolated from inanimate environments, such as sinks, floors, and bedsheets, in the rooms of the patients with MRSA infections as well as from the patients' hands. An epidemiological study demonstrated that clinical isolates of MRSA in our ward were of one origin and that the isolates from the air and from inanimate environments were identical to the MRSA strains that caused infection or colonization in the inpatients. CONCLUSIONS: Methicillin-resistant S aureus was recirculated among the patients, the air, and the inamimate environments, especially when there was movement in the rooms. Airborne MRSA may play a role in MRSA colonization in the nasal cavity or in respiratory tract MRSA infections. Measures should be taken to prevent the spread of airborne MRSA to control nosocomial MRSA infection in hospitals. PMID- 11405863 TI - Streptococcus milleri: an organism for head and neck infections and abscess. AB - BACKGROUND: Streptococcus milleri, a commensal organism, has the potential to cause significant morbidity. There is a paucity of published data regarding this organism in the head and neck. OBJECTIVES: To identify and assess the presentation, treatment, and outcomes of pediatric patients affected by this pathogen. STUDY DESIGN: Review of the Department of Pathology database at Children's Hospital of Wisconsin, Milwaukee, between 1997 and 1999 identified 26 patients with cultures positive for S milleri group (SMG) bacteria. Retrospective chart analysis examined the demographic data, site of origin of infection, additional organisms cultured, symptoms, treatments, and complications. RESULTS: Sixteen patients had SMG infections involving the head and neck region. Sites of origin included the paranasal sinuses, dental, facial soft tissues, deep neck spaces, peritonsillar region, and a tracheostomy site. The paranasal sinuses were the most common site in 37% (6/16). Streptococcus milleri was the only isolate in 69% (11) of the infections. Significant local extension occurred in 56% (9/16) of the patients and included the orbit, skull base, cranium, and deep neck spaces. All patients had surgical drainage and 15 also received intravenous antibiotic treatment. One complication of osteomyelitis of the frontal bone occurred with resolution after surgical debridement and intravenous antibiotic treatment. CONCLUSIONS: Streptococcus milleri can be an aggressive pathogen in the head and neck with a propensity for abscess formation and local extension of the infection in a pediatric population. Surgical drainage with antibiotics is generally successful in management of the condition. However, emerging penicillin resistance and the ability for local extension require suspicion of incomplete treatment if clinical symptoms persist. PMID- 11405864 TI - Chronic bacterial rhinosinusitis: description of a mouse model. AB - OBJECTIVES: To survey normal murine sinonasal anatomy and to create a mouse model for chronic bacterial rhinosinusitis. DESIGN: Anatomic, histologic, and pathophysiologic study displaying normal murine sinonasal anatomy and surgically created unilateral sinonasal inflammation. SUBJECTS: Twenty-one 6-week-old, male C57BL/6 mice. INTERVENTIONS: Animals that underwent unilateral maxillary sinus ostial obstruction using Merocel nasal packing, animals with unilateral Bacteroides fragilis inoculation alone, and animals with both ostial obstruction and bacterial inoculation were examined at 4 weeks for histologic evidence of chronic sinonasal inflammation. Experimental interventions were compared with contralateral control sinuses within each animal and with normal and sham operated controls. RESULTS: Normal mouse paranasal sinuses include maxillary sinuses, ethmoid air cells, and respiratory-type epithelium. In experimental animals, the lateral maxillary sinus wall, nasal septum, and superior turbinelle of the maxillary sinus were examined histologically. Epithelial thickening and disarray, goblet cell hyperplasia, inflammatory infiltrates, and sinonasal fibrosis were present in the experimental sinuses of animals packed with Merocel alone or Merocel with bacterial inoculation. Changes seen with Merocel and bacteria were more dramatic than those with Merocel alone. Sham-operated controls and sinuses inoculated with bacteria alone did not differ significantly from the sinuses of normal animals. CONCLUSION: Unilateral maxillary sinus ostial obstruction using Merocel nasal packing along with B fragilis inoculation results in a persistent, localized bacterial rhinosinusitis in mice. PMID- 11405865 TI - Titanium mesh repair of the severely comminuted frontal sinus fracture. AB - BACKGROUND: Severely comminuted frontal sinus fractures are difficult to contour and immobilize. Frequently, plates or wires are inadequate in fixating all fragments together, resulting in less than optimal outcomes. Advancements in the development of biomaterials have now made titanium mesh a new option for the repair of severely comminuted fractures. METHODS: Fourteen patients with severely comminuted frontal sinus fractures were treated with titanium mesh from 1994 to 1999. The fractures were reduced and immobilized using a simple algorithm: (1) Isolated anterior table fractures were repaired with reduced bony fragments attached to titanium mesh. (2) Anterior table fractures with nasofrontal duct involvement were repaired by sinus obliteration and anterior wall reconstruction with reduced bony fragments attached to titanium mesh. (3) Anterior and posterior table fractures with cerebrospinal fluid leak or displacement were treated with the cranialization of the sinus and anterior wall reconstruction with reduced bony fragments attached to titanium mesh. RESULTS: Of the 14 patients treated, 12 were available for postoperative evaluation. Parameters such as nasal function, cranial nerve V and VII function, cosmesis, and complications (hardware extrusions, sinusitis, meningitis, osteomyelitis, mucopyocele, brain abscess, pneumocephalus, and cerebrospinal fluid leak) were evaluated. All patients had good function of the superior division of cranial nerves V and VII. Two patients (16%) had minor wound infections, which resolved under treatment with antibiotics. All had excellent cosmetic results as measured by postreduction radiographs and personal and family perceptions of forehead contour. CONCLUSION: Titanium mesh reconstruction of severely comminuted frontal sinus fractures has few complications while providing excellent forehead contour and cosmesis. PMID- 11405866 TI - Quality-of-life outcomes in the evaluation of head and neck cancer treatments. AB - OBJECTIVES: To review the published literature to evaluate the design, use of terminology, and interpretation of results in studies using quality-of-life (QOL) instruments to measure differences between head and neck cancer treatments at a point in time or to report changes over time in one or more treatment groups. DATA SOURCE: MEDLINE search for subject headings "head and neck neoplasms" (as a main topic) and "quality of life" or "health status" restricted to English language sources and a 10-year period from 1989 to 1999. STUDY SELECTION: Four hundred forty-five abstracts were reviewed to find articles using an instrument to compare head and neck cancer therapy groups with a QOL outcome (13.7% included). DATA EXTRACTION: Two readers reviewed each article to determine how terminology was used, if a scientific study design was used, and if differences or changes in scores were clinically interpreted. RESULTS: Sixty-one articles were reviewed. Forty different instruments were used. Terminology was used inconsistently in 21 (34.4%) of the 61 articles. A scientific study design was used in only 11 (18.0%) of the 61 articles (P<.001). A clinical interpretation of results was given in 16 (26.2%) of the 61 articles (P<.001). CONCLUSIONS: While QOL outcomes show promise for assisting with treatment decisions in head and neck cancer therapy, few studies using instruments to measure QOL outcomes are hypothesis driven and clinical interpretations of results are not commonly provided. We recommend that future studies identify the construct to be measured, specify comparator groups and hypotheses a priori, and provide clinical interpretations of results. PMID- 11405867 TI - Urokinase-type plasminogen activator expression and proliferation stimulation in head and neck squamous cell carcinoma in vitro and in situ. AB - BACKGROUND: Stimulation of proliferative activity by urokinase-type plasminogen activator (uPA) has been demonstrated in vitro for cultured primary and carcinoma cells. OBJECTIVE: To examine the effect of uPA stimulation on cultured squamous cell carcinoma cell lines of the head and neck in vitro and to compare the results with the situation in tumor tissue specimens. DESIGN: The uPA-mediated growth stimulation of 2 head and neck squamous cell carcinoma cell lines after suppression of endogenous uPA production was monitored by measuring (3)H thymidine uptake into cellular DNA. Alternatively, applications of antibodies against the uPA-binding domain of the urokinase receptor were used to suppress autostimulation. To analyze the situation in situ we performed Western blot and zymographic studies on tissue homogenates of 25 squamous cell carcinoma specimens. We tested the expression of proliferating cell nuclear antigen (PCNA), a marker for proliferative activity, and uPA in tissue lysates and correlated uPA and PCNA expression by regression analysis. RESULTS: High-molecular-weight urokinase had a proliferation stimulative effect on both cell lines in vitro. The uPA autostimulation was decreased by blocking the uPA-binding domain of urokinase receptor with antibodies. Regression analysis of zymographic and Western blot data of tumor tissue lysates revealed no significant coherency between PCNA and uPA expression. Immunohistochemical stainings frequently showed different sublocalization of uPA and PCNA within tumors. CONCLUSION: In vitro uPA-mediated growth stimulation is not necessarily transferable to the in situ situation. PMID- 11405868 TI - High tumor grade in salivary gland mucoepidermoid carcinomas and loss of expression of transforming growth factor beta receptor type II. AB - BACKGROUND: Mucoepidermoid carcinoma (MEC) of salivary glands is a malignant, locally aggressive neoplasm with metastatic potential. The clinical course is usually dependent on histology; however, low-grade carcinomas can result in metastases and tumor-related death. Transforming growth factor beta1 (TGF-beta1) is a potent cytokine that affects growth inhibition of various cells and stimulates extracellular matrix production and angiogenesis. Loss of TGF-beta receptor type II (TGF-beta RII) expression has been related to resistance of TGF beta1-mediated growth control and tumor progression. In this study, we correlate MEC tumor grade with expression of TGF-beta1 and TGF-beta RII. DESIGN: Immunohistochemical staining was performed on 16 MEC specimens for activated forms of TGF-beta1 and TGF-beta RII. The percentage of cells in which staining yielded positive findings for activated TGF-beta1 and TGF-beta RII was correlated with tumor grade. RESULTS: Activated TGF-beta1 was detected in 16 specimens (100%) of MEC and showed strong positive and diffuse staining. Predominately cytoplasmic staining of TGF-beta1 was seen in salivary gland ducts, stroma, and endothelial cells. There was an inverse correlation between tumor grade and loss of expression of TGF-beta RII. All low-grade MEC tumors yielded positive staining results, whereas only one case of intermediate-grade MEC had TGF-beta RII expression. No high-grade MEC showed TGF-beta RII expression. CONCLUSIONS: Loss of expression of TGF-beta RII correlates with tumor grade. The localization of activated TGF-beta1 within neoplastic epithelium, tumor-associated stroma, and endothelium suggests that it might play a role in the stromal proliferation and/or angiogenesis associated with MEC. PMID- 11405869 TI - The transglabellar/subcranial approach to the anterior skull base: a review of 72 cases. AB - OBJECTIVES: To describe the transglabellar/subcranial approach to the anterior skull base and to compare it with more traditional approaches to craniofacial resection. DESIGN: A retrospective analysis of 72 cases at 2 academic medical centers. The main parameters analyzed were the disease entities treated, the average operating room time, the average amount of blood loss, the number of transfusions, the length of intensive care unit and hospital stays, and complication rates. These were compared with published data for traditional craniofacial approaches. SETTING: All patients were operated on by the authors in collaboration with neurosurgical teams at the State University of New York Upstate Medical University, Syracuse, and the University of Michigan Hospital, Ann Arbor. PATIENTS: The transglabellar/subcranial approach was performed 72 times in 69 patients in this series. Forty-two procedures in 40 patients were performed for malignant disease and 30 procedures in 29 patients were performed for benign entities. Patients' ages ranged from 2 to 78 years. Follow-up ranged from 6 months to 4 years, with a minimum follow-up of 1 year for survivors. RESULTS: There were no operative mortalities. Operating time, average amount of blood loss, length of hospital and intensive care unit stays, and complication rates compared favorably with published results of traditional craniofacial resections. CONCLUSIONS: The transglabellar/subcranial approach to the anterior skull base may be a reasonable technique for the surgical management of lesions in the region of the anterior skull base. It provides excellent exposure of the nasal cavity, the orbits, and the ethmoid and sphenoid sinuses, while allowing wide access to the anterior fossa with a minimum amount of frontal lobe retraction. PMID- 11405870 TI - Endoscopic laser cricopharyngeal myotomy to salvage tracheoesophageal voice after total laryngectomy. AB - Development of voice after tracheoesophageal puncture, following laryngectomy, is sometimes hampered by spasm of the cricopharyngeal muscle. This problem has been addressed by various means, including bougienage, botulinum toxin injection, and open surgical division of the muscle. We believe that endoscopic carbon dioxide laser cricopharyngeal myotomy represents a direct, simple, and effective solution. PMID- 11405871 TI - Unilateral vs. bilateral supraglottoplasty for severe laryngomalacia in children. AB - OBJECTIVES: To study the efficacy of unilateral supraglottoplasty in comparison with bilateral supraglottoplasty for the treatment of severe laryngomalacia in children and to study factors that may be predictive of major complications or the need for a subsequent contralateral or revision procedure. DESIGN: Retrospective medical record review. SETTING: University tertiary care pediatric hospital. PATIENTS: One hundred six consecutive pediatric patients, aged 9 days to 18 years, who had undergone unilateral or bilateral supraglottoplasty for severe laryngomalacia. MAIN OUTCOME MEASURES: Resolution of clinically significant laryngomalacia, development of major complications (supraglottic stenosis or aspiration), and an association between study variables (demographics, medical comorbidities, synchronous airway abnormalities, sites of excision, and techniques of excision) and the need for subsequent contralateral or revision supraglottoplasty. RESULTS: We achieved a high success rate (95.7%), a low complication rate (8.5%), and observed the need for a contralateral procedure in 7 (14.9%) of the 47 patients who underwent initial unilateral supraglottoplasty. Two patients who underwent initial bilateral supraglottoplasty developed supraglottic stenosis. No significant association existed between our study variables and the development of complications or the need for contralateral or revision supraglottoplasty. CONCLUSIONS: Unilateral supraglottoplasty was associated with a high success rate, low complication rate, and the avoidance of supraglottic stenosis in our study population. The percentage of patients requiring a subsequent contralateral procedure was comparable to that reported in the literature, and no major complications were associated with the second operation in these patients. Therefore, unilateral supraglottoplasty seems to be a reasonable option for initial surgical management of pediatric patients with severe laryngomalacia. PMID- 11405872 TI - Aerodynamic findings in esophageal voice. AB - OBJECTIVE: To define the perceptive and aerodynamic characteristics of esophageal voice in relation to different rehabilitation modalities. DESIGN: Cross-sectional study comparing perceptive and aerodynamic variables in 3 subject groups. SETTING: Referral center. SUBJECTS: A total of 19 subjects who underwent total laryngectomy were divided into groups A and B. Group A consisted of 13 subjects (who required speech therapy)-8 good speakers (subset A(1) who were >80% intelligible) and 5 mediocre speakers (subset A(2) who were <70% intelligible). Group B consisted of 6 subjects with a tracheoesophageal prosthesis (who were >90% intelligible). MAIN OUTCOME MEASURES: Perceptive variables included phonatory pauses and stomal noise. Aerodynamic variables included maximum phonation time, phonatory flow, phonatory volume, postphonatory volume, intensity, and articulatory pressure. RESULTS: Phonatory pauses and stomal noise statistically differentiated group A from group B and good speakers from mediocre speakers. Phonation time, phonatory volume, and phonatory flow were statistically higher in group B subjects compared with group A subjects. Postphonatory volume was significantly higher in group A. Intraoral pressure and postphonatory volume were statistically higher in subset A(2) subjects compared with subset A(1) subjects while maximum phonation time was significantly higher in subset A(1) subjects compared with subset A(2) subjects. CONCLUSIONS: In subset A(1) subjects a positive ratio between phonatory volume and phonatory flow was maintained with an adequate phonation time. In subset A(2) subjects a reduced phonatory volume was associated with a more rapid dispersion of phonatory flow, lower duration of phonation, and frequent pauses; stomal noise and consonant hyperarticulation worsened the voice performance in this group. In group B subjects the positive ratio between phonatory volume and phonatory flow represented the prerequisite of speech without frequent pauses. PMID- 11405873 TI - Long-term audiological feature in Pendred syndrome caused by PDS mutation. AB - Pendred syndrome is an autosomal recessive disorder characterized by profound deafness in childhood and goiter. We report a case of Pendred syndrome in a 27 year-old woman who had a diffuse goiter and progressive sensorineural hearing loss with fluctuation and a missense mutation (His723Arg) in the PDS gene identified in a homozygous state. Audiological findings were observed clinically over a 20-year period. Progressive hearing loss with fluctuation occurred before age 12 years. An enlarged vestibular aqueduct with enlargement of the endolymphatic duct and sac was confirmed with 3-dimensional magnetic resonance imaging hydrography. PMID- 11405874 TI - Nasal cavity squamous cell carcinoma in Wegener's granulomatosis. AB - Wegener's granulomatosis is a chronic debilitating disease with multiple organ system involvement, variable course, and myriad complications that cause morbidity and mortality. We report 2 cases of nasal cavity squamous cell carcinoma occurring in long-standing Wegener's granulomatosis. PMID- 11405875 TI - Radiology forum: quiz case 1. Diagnosis: superficial siderosis. PMID- 11405876 TI - Radiology forum: quiz case 2. Diagnosis: petrous apicitis with secondary abducens nerve palsy. PMID- 11405877 TI - Should formal ophthalmologic evaluation be a preoperative requirement prior to blepharoplasty? PMID- 11405878 TI - Ophthalmic evaluation should be a preoperative requirement prior to blepharoplasty. PMID- 11405879 TI - Preoperative ophthalmic evaluation is a personal choice. PMID- 11405880 TI - Role of transcription factor NFAT in the immune response. AB - Molecular mechanisms of activation of nuclear factor NFAT in cells of the lympho myeloid complex are considered. Members of the NFAT family regulate transcription of genes encoding proteins involved in the induction and/or regulation of the immune response. It is possible that altered transcription activity of NFAT under conditions of its deficit or blockade of expression may account for changes in the immune status of an organism. PMID- 11405881 TI - Oxidative stress and mechanisms of protection against it in bacteria. AB - In the review contemporary data on the effects of oxidative stresses of various kinds in bacteria are summarized. A general theory of oxidative stress, peculiarities of oxidative stress in eukaryotes and prokaryotes, and natural and induced oxidative stresses are described. Data on the mechanisms of protection against oxidative stress are given, including prevention of the generation of oxidative stress, prevention of propagation of free radical chain reactions, and the mechanisms of repair of damaged DNA. The regulation of effector genes via redox-sensitive iron-containing proteins is analyzed. Special attention is given to the expression of so-called antioxidant and associated enzymes as protection mechanisms and to the space-time organization of the response of bacteria to oxidative stress. PMID- 11405882 TI - Light-induced biogenesis of chlorophyll-protein complexes in developing wheat thylakoids. AB - It is shown that the apoproteins of core complexes (CC) I and II, the alpha- and beta-subunits of CF1 ATP-synthase complexes, are present in seedlings grown under intermittent light (IML). The levels of light-harvesting complex (LHC) apoproteins in the 30- to 18-kD region increase rapidly upon exposure to continuous light (CL). The newly synthesized LHC apoproteins appear to be present predominantly in monomeric forms that later assemble into higher-order oligomeric forms. During the early stages of greening of wheat seedlings, polypeptides in the 20.5-19 and 17.5-15.5 kD regions (so-called "early light-induced proteins" (ELIP)) are observed, but they disappear fully after 6 h. As greening proceeds, the 727-nm band in low-temperature fluorescence spectra (77 K) gradually shifts to longer wavelength (740-742 nm), which clearly demonstrates the light-driven biogenesis of LHC I and its assembly with CC I. PMID- 11405883 TI - Induction of a peroxisomal malate dehydrogenase isoform in liver of starved rats. AB - The influence of starvation on malate dehydrogenase (MDH) in rat liver was investigated. Native electrophoresis revealed two MDH isoforms in non-starved rats and three isoenzymes in starved rats. After sucrose density gradient centrifugation of cell organelles from liver, MDH activity was detected in the mitochondrial and cytosolic fractions from non-starved rats. However, additional activity was found in the peroxisomal fraction from starved rats. The latter was identified as the electrophoretically new isoform in starved animals. The three isoforms of malate dehydrogenase from hepatocytes were separated and partially purified by chromatography on DEAE-Toyopearl. Several kinetic and regulatory properties of the three isoforms were rather similar. It is suggested that the newly expressed isoform of MDH operates in the glyoxylate cycle of liver peroxisomes of food-starved animals. PMID- 11405884 TI - Sphinganine in sphingomyelins of tumors and mouse regenerating liver. AB - Contents of sphingenine (sphingosine) and sphinganine were studied in sphingomyelins of transplantable mouse tumors (hepatoma-22, melanoma B16, Lewis lung carcinoma, intestine carcinoma) and rat nephroma RA. The content of sphinganine was increased in sphingomyelins of hepatoma-22 and nephroma RA compared to sphingomyelins of liver and kidneys. Significant contents of sphinganine were also found in sphingomyelins of other studied tumors. The content of sphinganine in regenerating mouse liver (30 h after hepatectomy) was normal. The data suggest that disorders should exist in biosynthesis of sphingoid bases in tumors but not in normal rapidly proliferating tissue. PMID- 11405885 TI - Initiation and inhibition of free radical processes in H2O2-metmyoglobin (methemoglobin)-2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) systems. AB - Rates of free radical initiation were determined at 20 degrees C in 10 mM phosphate buffer (pH 7.4) in the systems metmyoglobin (methemoglobin)-H2O2 using 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) as the diammonium salt (ABTS). The catalytic activity of MetMb was 2-3-fold higher than that of MetHb. The process can be described by the Michaelis-Menten equation, from which effective values of Km and Vmax were calculated. Comparative kinetic studies on the inhibition of ABTS oxidation were carried out using Trolox, propylgallate (PG), polydisulfide of gallic acid (poly(DSG)), polydisulfide of (2-amino-4 nitrophenol) (poly(ADSNP)), and its conjugate with human serum albumin (HSA poly(ADSNP)). The inhibitors were characterized by inhibition constants Ki and stoichiometric inhibition coefficients f (the number of radicals terminated by a single molecule of inhibitor). The minimum Ki and the maximum f values were obtained for poly(DSG), and in the system of MetHb-H2O2-ABTS they were 0.08 microM and 27.5, respectively. According to their antiradical activities, the inhibitors can be arranged as follows: poly(DSG) > poly(ADSNP) > PG > Trolox. PG, poly(DSG), poly (ADSNP), and its conjugate with HSA are suggested as "calibrators", i.e., inhibition standards for evaluation of antioxidant status of biological fluids in possible test systems based on the free radical-generating pair MetMb-H2O2 with ABTS as the acceptor of the active radicals. PMID- 11405886 TI - Long-term chemiluminescent signal is produced in the course of luminol peroxidation catalyzed by peroxidase isolated from leaves of african oil palm tree. AB - Optimal conditions were found for the oxidation of luminol by hydrogen peroxide in the presence of peroxidase isolated from leaves of the African oil palm tree Elaeis guineensis (AOPTP). The pH range for maximal chemiluminescence intensity (8.3-8.6) is similar for AOPTP, horseradish, and Arthromyces ramosus peroxidases and slightly different from that for tobacco peroxidase (9.3). Increasing the buffer concentration decreases the chemiluminescence intensity. As in the case of other anionic peroxidases, the catalytic efficiency of AOPTP does not depend on the presence of enhancers (4-iodophenol and 4-hydroxycinnamic acid) in the reaction medium. The detectable limit of AOPTP assayed by luminol peroxidation is 2.10(-12) M. The long-term chemiluminescence signal produced during AOPTP dependent luminol peroxidation is a characteristic feature of the African oil palm enzyme. This feature in combination with its very high stability suggests that AOPTP will be a promising tool in analytical practice. PMID- 11405887 TI - Comparative study of thermal degradation of iron-sulfur proteins in spinach chloroplasts and membranes of thermophilic cyanobacteria: mossbauer spectroscopy. AB - Mossbauer spectra of chloroplasts isolated from spinach plants grown in a mineral medium enriched with 57Fe and Mossbauer spectra of native membranes of the thermophilic cyanobacterium Synechococcus elongatus contain a broad asymmetric doublet typical of the iron-sulfur proteins of Photosystem (PS) I. Exposure of chloroplasts to temperatures of 20-70 degrees C significantly modifies the central part of the spectra. This spectral change is evidence of decreased magnitude of the quadrupole splitting. However, the thermally induced doublet (DeltaQ = 3.10 mm/sec and delta = 1.28 mm/sec) typical of hydrated forms of reduced (divalent) inorganic iron is not observed in spinach chloroplasts. This doublet is usually associated with degradation of active centers of ferredoxin, a surface-exposed protein of PS I. The Mossbauer spectra of photosynthetic membranes of spinach chloroplasts and cyanobacteria were compared using the probability distribution function of quadrupole shift (1/2 quadrupole splitting DeltaQ) of trivalent iron. The results of calculation of these functions for the two preparations showed that upon increasing the heating temperature there was a decrease in the probability of the presence of native iron-sulfur centers FX, FA, and FB (quadrupole shift range, 0.43-0.67 mm/sec) in heated preparations. This process was also accompanied by an increase in the probability of appearance of clusters of trivalent iron. This increase was found to be either gradual and continuous or abrupt and discrete in photosynthetic membranes of cyanobacteria or spinach chloroplasts, respectively. The probability of the presence of the iron sulfur centers FX, FA, and FB in chloroplasts abruptly decreases to virtually to zero within the temperature range critical for inhibition of electron transport through PS I to oxygen. In cyanobacteria, both thermal destruction of iron-sulfur centers of PS I and functional degradation of PS I are shifted toward a higher temperature. The results of this study suggest that the same mechanism of thermal destruction of the PS I core occurs in both thermophilic and mesophilic organisms: destruction of iron-sulfur centers FX, FA, and FB, release of oxidized (trivalent) iron, and its accumulation in membrane-bound iron-oxo clusters. PMID- 11405888 TI - Epitope mapping of human alpha-fetoprotein. AB - The epitope structure of human alpha-fetoprotein (AFP) was studied using more than 50 monoclonal antibodies (MAB) to human AFP. These MAB obtained from various world laboratories of the TD-2 AFP Workshops of the International Society for Oncodevelopmental Biology and Medicine (ISOBM-1996-1998-2000) were analyzed by competitive immunoaffinity electrochromatography (IAE) on nitrocellulose membranes (NCM). Five types of interaction of the AFP-MAB complex with the MAB fixed on NCM were found: 1) complete neutralization; 2) partial neutralization; 3) unidirectional neutralization; 4) enhanced binding; 5) lack of interaction. By IAE, 51 MAB were found to recognize 23 different epitopes in the AFP molecule. Based on these findings, an epitope map of AFP was designed which consists of eight epitope clusters and eight individual epitopes. The epitope location is considered with respect to the conformational state of the AFP molecule. Possible causes of the five types of interaction found on neutralization are discussed. PMID- 11405889 TI - Use of trinitrophenylation for quantification of protease and peptidase activities. AB - A sensitive and precise method for quantifying protease and peptidase activities is suggested. N-Terminal amino groups of peptides which are formed during hydrolysis of the substrates react with trinitrobenzenesulfonic acid (TNBS), and the trinitrophenyl (TNP) derivatives are determined spectrophotometrically. Spontaneous hydrolysis of TNBS is considerably diminished on trinitrophenylation at pH 7.4 rather than at pH 9-10 as is usually used. The trinitrophenylation method can be used to determine the initial rate of hydrolysis and the kinetics of reactions catalyzed by proteases and peptidases. PMID- 11405890 TI - American cockroach (Periplaneta americana) synthesizes carotenoids from the precursor. AB - The level of an important carotenoid (beta-carotene) in the gut of Periplaneta americana depends on the content of the carotenoid in food: a carotenoid fortified diet causes accumulation of beta-carotene up to 10 microg/g wet weight, while on a carotenoid-deficient diet the level of this substance is low (approximately 0.7 microg/g wet weight). In the eye, in contrast to the gut, a constant level of beta-carotene (1.3-1.4 microg/g wet weight) is found regardless of the diet. This phenomenon remained unchanged over three years of feeding of the cockroaches with the carotenoid-deficient diet, suggesting that P. americana produces carotenoids by de novo biosynthesis. This suggestion was confirmed in experiments using intraperitoneal injection of the exogenous carotenoid biosynthesis precursor [14C]mevalonic acid pyrophosphate followed by extraction of carotenoid and chromatographic purification of the labeled product. Injection of 3.4 nmoles [14C]mevalonic acid pyrophosphate transiently increased the beta carotene content in eyes on days 2 and 4 after injection of the label. Purification of radiolabeled carotenoids from eye and gut by the transfer of carotenoids into a less polar solvent, alkaline hydrolysis (saponification), and chromatography on alumina and cellulose columns decreased the specific radioactivity to a constant level that cannot be further decreased by repeated chromatography. The elution profile of these purified preparations of beta carotene after chromatography is characterized by coincidence of symmetric peaks of count and absorption. We suggest that to create the optimal carotenoid concentration in the eye, P. americana uses two biochemical mechanism: 1) it accumulates carotenoids in reserve in the gut when abundant supplies of carotenoids are available in the diet; 2) it synthesizes carotenoids de novo when its food is deficient in these compounds. PMID- 11405891 TI - Studies of electron transport dynamics in photosynthetic reaction centers using fast temperature changes. AB - Rates of thermoinduced conformational transitions of reaction center (RC) complexes providing effective electron transport were studied in chromatophores and isolated RC preparations of various photosynthesizing purple bacteria using methods of fast freezing and laser-induced temperature jump. Reactions of electron transfer from the primary to secondary quinone acceptors and from the multiheme cytochrome c subunit to photoactive bacteriochlorophyll dimer were used as probes of electron transport efficiency. The thermoinduced transition of the acceptor complex to the conformational state facilitating electron transfer to the secondary quinone acceptor was studied. It was shown that neither the characteristic time of the thermoinduced transition within the temperature range 233-253 K nor the characteristic time of spontaneous decay of this state at 253 K exceeded several tens of milliseconds. In contrast to the quinone complex, the thermoinduced transition of the macromolecular RC complex to the state providing effective electron transport from the multiheme cytochrome c to the photoactive bacteriochlorophyll dimer within the temperature range 220-280 K accounts for tens of seconds. This transition is thought to be mediated by large-scale conformational dynamics of the macromolecular RC complex. PMID- 11405892 TI - Enzymes of a new modified ortho-pathway utilizing 2-chlorophenol in Rhodococcus opacus 1CP. AB - Chlorocatechol 1,2-dioxygenase (CC 1,2-DO), chloromuconate cycloisomerase (CMCI), chloromuconolactone isomerase (CMLI), and dienolactone hydrolase (DELH), the key enzymes of a new modified ortho-pathway in Rhodococcus opacus 1CP cells utilizing 2-chlorophenol via a 3-chlorocatechol branch of a modified ortho-pathway, were isolated and characterized. CC 1,2-DO showed the maximum activity with 3 chlorocatechol; its activity with catechol and 4-chlorocatechol was 93 and 50%, respectively. The enzyme of the studied pathway had physicochemical properties intermediate between the pyrocatechase of ordinary and chlorocatechase of modified pathways described earlier for this strain. In contrast to the enzymes investigated earlier, CMCI of the new pathway exhibited high substrate specificity. The enzyme had Km for 2-chloromuconate of 142.86 microM, Vmax = 71.43 U/mg, pH optimum around 6.0, and temperature optimum at 65 degrees C. CMCI converted 2-chloromuconate into 5-chloromuconolactone. CMLI converted 5 chloromuconolactone into cis-dienolactone used as a substrate by DELH; this enzyme did not convert trans-dienolactone. DELH had Km for cis-dienolactone of 200 microM, Vmax = 167 U/mg, pH optimum of 8.6, and temperature optimum of 40 degrees C. These results confirm the existence of a new modified ortho-pathway for utilization of 2-chlorophenol by R. opacus 1CP. PMID- 11405893 TI - Ion-exchange properties of cell walls isolated from lupine roots. AB - Acid-base properties of cell walls isolated from various root tissues of 7-day old lupine seedlings and 14-day-old lupine plants grown in various media were studied. The ion-exchange capacity of root cell walls was estimated at various pH values (from 2 to 12) and constant ionic strength (10 mM). The parameters determining the qualitative and quantitative composition of cell wall ionogenic groups along the root length and in its radial direction were estimated using Gregor's model. This model fits the experimental data reasonably well. Four types of ionogenic groups were found in the cell walls: an amino group (pKa approximately 3), two types of carboxylic groups (pKa approximately 5 and 7.3, the first being the carboxylic group of galacturonic acid), and a phenolic group (pKa approximately 10). The number of functional groups of each type was estimated, and the corresponding ionization constant values were calculated. It is shown that the chemical composition of the ionogenic groups was constant along the root length as well as in its radial direction and did not depend on either physiological state or root nutrition, while the number of different groups varied. The content of carboxylic groups of alpha-D-polygalacturonic acid in the root cell walls of 14-day-old plants was shown to depend on the distance from the root tip, being maximal in the zone of lateral roots. The number of these groups was 10- and 2-fold less in the central cylinder compared to that of cortex for 14 day-old plants and 7-day-old seedlings, respectively. PMID- 11405894 TI - Site-directed mutagenesis of cytochrome P450scc. II. Effect of replacement of the Arg425 and Arg426 residues on the structural and functional properties of the cytochrome P450scc. AB - Cytochrome P450-dependent monooxygenases, in spite of their wide distribution, can be simply divided into a few groups differing in the location of the electron transfer chain and their composition. The two main groups of cytochrome P450 dependent monooxygenases are the mitochondrial and microsomal enzymes. While in two-component microsomal cytochrome P450-dependent monooxygenases electrons are supplied to cytochrome P450 by a flavoprotein (NADPH-cytochrome P450 reductase), in three-component mitochondrial monooxygenases the electrons are supplied to cytochrome P450 by a low molecular weight protein (ferredoxin). The interaction of cytochrome P450 with NADPH-cytochrome P450 reductase and ferredoxin is the subject of intensive studies. Using chemical modification, chemical cross linking, and site-directed mutagenesis, we identified surface exposed positively charged residues of cytochrome P450scc which might be important for interaction with adrenodoxin. Theoretical analysis of the distribution of surface electrostatic potential in cytochrome P450 indicates that in contrast to microsomal monooxygenases, cytochromes P450 of mitochondrial type, and cholesterol side-chain cleavage cytochrome P450 (P450scc) in part, carry on the proximal surface an evidently positively charged site that is formed by residues Arg425 and Arg426. In the present work, to estimate the functional role of Arg425 and Arg426 of cytochrome P450scc, we used site-directed mutagenesis to replace these residues with glutamine. The results indicate that residues Arg425 and Arg426 are involved in the formation of a heme-binding center and electrostatic interaction of cytochrome P450scc with its physiological electron-transfer partner, adrenodoxin. PMID- 11405895 TI - Changes in antioxidative properties of lactoferrin from women's milk during deamidation. AB - Spontaneous deamidation of lactoferrin preparations from women's milk was found during incubation for 28 days under simulated physiological conditions (0.85% NaCl, pH 7.0, 37 degrees C). After 28 days of incubation, this deamidation was associated with a 12% decrease in the total amide content in the protein. Addition of deamidated preparation to a suspension of lipoproteins from egg yolk in the presence of Rhodamine 6G decreased the total intensity of rapid and slow emission and also the intensity of the slow emission of Fe2+-induced chemiluminescence by 37, 48, and 53%, respectively, suggesting an increase in the antioxidative activity of lactoferrin during deamidation. Deamidation obviously stimulated the nonspecific interaction of lactoferrin with iron ions and, consequently, increased the antioxidant effect of the protein in this way. This was supported by the finding of decreased antioxidative effectiveness of lactoferrin during its complete saturation with iron under the incubation conditions. PMID- 11405896 TI - Reporting of conflicts of interest in guidelines of preventive and therapeutic interventions. AB - BACKGROUND: Guidelines published in major medical journals are very influential in determining clinical practice. It would be essential to evaluate whether conflicts of interests are disclosed in these publications. We evaluated the reporting of conflicts of interest and the factors that may affect such disclosure in a sample of 191 guidelines on therapeutic and/or preventive measures published in 6 major clinical journals (Annals of Internal Medicine, BMJ, JAMA, Lancet, New England Journal of Medicine, Pediatrics) in 1979, 1984, 1989, 1994 and 1999. RESULTS: Only 7 guidelines (3.7%) mentioned conflicts of interest and all were published in 1999 (17.5% (7/40) of guidelines published in 1999 alone). Reporting of conflicts of interest differed significantly by journal (p=0.026), availability of disclosure policy by the journal (p=0.043), source of funding (p < 0.001) and number of authors (p=0.004). In the entire database of 191 guidelines, a mere 18 authors disclosed a total of 24 potential conflicts of interest and most pertained to minor issues. CONCLUSIONS: Despite some recent improvement, reporting of conflicts of interest in clinical guidelines published in influential journals is largely neglected. PMID- 11405897 TI - In silico discovery of novel retinoic acid receptor agonist structures. AB - BACKGROUND: Several Retinoic Acid Receptors (RAR) agonists have therapeutic activity against a variety of cancer types; however, unacceptable toxicity profiles have hindered the development of drugs. RAR agonists presenting novel structural and chemical features could therefore open new avenues for the discovery of leads against breast, lung and prostate cancer or leukemia. RESULTS: We have analysed the induced fit of the active site residues upon binding of a known ligand. The derived binding site models were used to dock over 150,000 molecules in silico (or virtually) to the structure of the receptor with the Internal Coordinates Mechanics (ICM) program. Thirty ligand candidates were tested in vitro. CONCLUSIONS: Two novel agonists resulting from the predicted receptor model were active at 50 nM. One of them displays novel structural features which may translate into the development of new ligands for cancer therapy. PMID- 11405898 TI - Systematic review of the use of honey as a wound dressing. AB - OBJECTIVE: To investigate topical honey in superficial burns and wounds though a systematic review of randomised controlled trials. DATA SOURCES: Cochrane Library, MEDLINE, EMBASE, PubMed, reference lists and databases were used to seek randomised controlled trials. Seven randomised trials involved superficial burns, partial thickness burns, moderate to severe burns that included full thickness injury, and infected postoperative wounds. REVIEW METHODS: Studies were randomised trials using honey, published papers, with a comparator. Main outcomes were relative benefit and number-needed-to-treat to prevent an outcome relating to wound healing time or infection rate. RESULTS: One study in infected postoperative wounds compared honey with antiseptics plus systemic antibiotics. The number needed to treat with honey for good wound healing compared with antiseptic was 2.9 (95% confidence interval 1.7 to 9.7). Five studies in patients with partial thickness or superficial burns involved less than 40% of the body surface. Comparators were polyurethane film, amniotic membrane, potato peel and silver sulphadiazine. The number needed to treat for seven days with honey to produce one patient with a healed burn was 2.6 (2.1 to 3.4) compared with any other treatment and 2.7 (2.0 to 4.1) compared with potato and amniotic membrane. For some or all outcomes honey was superior to all these treatments. Time for healing was significantly shorter for honey than all these treatments. The quality of studies was low. CONCLUSION: Confidence in a conclusion that honey is a useful treatment for superficial wounds or burns is low. There is biological plausibility. PMID- 11405899 TI - Life after pacemaker implantation: management of common problems and environmental interactions. AB - Patients with permanent pacemakers have become an increasingly common clinical entity for physicians. Many patients who receive pacemakers are active people who leave the hospital expecting to return to fully functional, normal lives. These patients need to be aware of potential pacemaker interactions at work and in other environments. One location in which many patients with pacemakers find themselves is the hospital environment, which is host to an abundant array of diagnostic and therapeutic sources of electromagnetic interference that are capable of disrupting normal pacemaker function. Previous studies have addressed issues ranging from these patients' use of cellular telephones to the dangers of magnetic resonance imaging. This article reviews the medical literature on the management of patients with permanent pacemakers and the problems that these patients may encounter because of electromagnetic interference. PMID- 11405900 TI - Obesity and cardiovascular disease risk: research update. AB - The obesity epidemic has reached unprecedented proportions in Western society. Evidence continues to accumulate that obesity is associated with significant morbidity and mortality and in particular that it is an independent risk factor for cardiovascular disease (CVD). The association of obesity with CVD and its risk factors, including hypertension, dyslipidemia, glucose intolerance, and impaired hemostasis is becoming more clearly understood. An increasing body of data indicates that risk factors tend to cluster in obese individuals and may act synergistically to increase these people's risk for CVD. Individuals with disproportionate visceral adiposity are at significantly greater risk for CVD. Adult weight gain also underlies the development of many risk factors and augments the risk of CVD. Physicians can play a vital and active role in the prevention and treatment of obesity and overweight and thereby reduce patients' CVD risk. PMID- 11405901 TI - Echocardiographic assessment of mitral regurgitation. AB - Although the natural history of mitral regurgitation (MR) is poorly defined, evidence has been found for excess mortality and morbidity in patients with severe MR who are managed conservatively. With improved mortality and morbidity in the surgical management of this condition, we are becoming increasingly aggressive in offering surgery to patients with severe MR. Surgery may be offered even in the absence of symptoms or left ventricular dysfunction, provided that the valve seems reparable, the patient's MR is severe, and the surgical team is experienced in valve repair. Echocardiography is critically important in determining the feasibility of valve repair and accurately assessing the severity of the patient's MR. It also allows assessment of the effect of MR on the left ventricle and the left atrium. PMID- 11405902 TI - Biventricular pacing therapy for congestive heart failure: a review of the literature. AB - Biventricular pacing is a new therapy for patients with congestive heart failure and mechanical dyssynchrony. Its therapeutic intent is to activate both ventricles simultaneously, thus improving the mechanical efficiency of the ventricles. Preliminary evidence indicates improvement in hemodynamics, quality of life, and exercise capacity in patients in sinus rhythm as well as in patients with atrial fibrillation. An improvement in diastolic filling, a decrease in mitral regurgitation, and more efficient systolic ejection are proposed as the mechanisms behind these benefits. In addition, some evidence indicates that the frequency of ventricular arrhythmias is decreased. All clinicians involved in the management of patients with congestive heart failure eagerly await the results of ongoing trials. The results of these trials will define which patients are eligible for this therapy, which patients will derive the most benefit from it, and its effect on morbidity and mortality. PMID- 11405903 TI - Diagnosis and treatment of severe pediatric pulmonary hypertension. AB - Advances in the treatment of pulmonary hypertension during the past decade have dramatically improved patient survival. Many of these advances are based on improved understanding of the vascular biology of the normal and hypertensive pulmonary circulations. Pulmonary hypertension is an important determinant of morbidity and mortality in patients with many pediatric diseases, including congenital heart disease. This article describes current diagnostic strategies and treatments for patients with primary and secondary pulmonary hypertension. PMID- 11405904 TI - Medical management of peripheral arterial disease. AB - Peripheral arterial disease affects approximately 8-10 million people in the United States. Approximately one-third to one-half of these individuals are symptomatic. The risk factors that contribute to peripheral arterial disease are similar to those associated with other forms of atherosclerosis, including diabetes mellitus, cigarette smoking, hypercholesterolemia, high blood pressure, and hyperhomocysteinemia. Of these, diabetes and cigarette smoking pose the greatest risk for developing peripheral arterial disease. The prognosis of patients with these risk factors is limited because of their greater risks for myocardial infarction, stroke, and cardiovascular death. Cardiovascular mortality correlates inversely with the ankle/brachial index, and the risk of death is greatest in those with the most severe peripheral arterial disease. Treatment regimens to reduce cardiovascular morbidity and mortality in patients with peripheral arterial disease should include risk factor modification and antiplatelet therapy. The cardinal symptoms of peripheral arterial disease include intermittent claudication and rest pain, with the latter being indicative of critical limb ischemia. Therapeutic strategies that focus on improving the patient's quality of life, reducing the severity of claudication, and improving limb viability include supervised exercise training, pharmacotherapy, and revascularization. Two drugs-pentoxifylline and cilostazol-currently are approved by the Food and Drug Administration for the treatment of patients with claudication. Meta-analyses have suggested that, compared with placebo, pentoxifylline improves maximal walking distance by approximately 20-25%. Cilostazol is a phosphodiesterase type 3 inhibitor. In clinical trials, cilostazol has consistently improved maximal walking distance as compared with placebo, with the range of improvement being approximately 40-60%. Drugs that are currently under investigation include propionyl-L-carnitine, vasodilator prostaglandins, L-arginine, and the angiogenic factors, vascular endothelial growth factor and basic fibroblast growth factors. PMID- 11405905 TI - Effects of endurance training on the breathing pattern of professional cyclists. AB - The aim of this longitudinal study was to clarify the changes induced by endurance training on the breathing pattern of 13 professional cyclists (age+/ SD: 24+/-2 years; VO(2 max) approximately 75 ml x kg(-1) x min(-1)) during the three periods (rest, precompetition, and competition) of a sports season. Both the volume and the intensity of training were quantified during these periods. In each session (corresponding to each of the three periods) all subjects performed (1) a pulmonary function test (to measure forced vital capacity [FVC], peak expiratory flow [PEF], and maximal voluntary ventilation [MVV]), and (2) a ramp test until exhaustion on a cycle ergometer (workload increases of 25 W x min( 1)). The following variables were recorded every 100 W until the end of the tests: pulmonary ventilation (VE, in l x min(-1) BTPS), tidal volume (VT, inl BTPS), breathing frequency (f(b), in breaths x min(-1)), ventilatory equivalents for oxygen (VE x VO(2)(-1)) and carbon dioxide (VE x VCO(2)(-1)), inspiratory (TI) and expiratory (TE) times (s), ratio of TI to total respiratory duration or inspiratory "duty cycle" (TI/TTOT, and mean inspiratory flow rate (VT)/TI), in l x s(-1)). The results showed no changes in any of these variables (p>0.05) between the three periods of study, despite significant changes in training loads (i.e., increases in the volume and/or intensity of training throughout the season). These findings suggest that endurance conditioning does not alter the breathing pattern of professional cyclists during an incremental exercise test. PMID- 11405906 TI - New calculation of internal Ca(2+) recirculation fraction from alternans decay of postextrasystolic potentiation. AB - In our previous studies, we calculated the internal Ca(2+) recirculation fraction (RF) after obtaining the beat decay constant (tau(e)) of the monoexponential component in the postextrasystolic potentiation (PESP) of the alternans decay by curve fitting. However, this method sometimes suffers from the sensitive variation of tau(e) with small noises in the measured contractilities of the 5th and 6th postextrasystolic (PES) beats in the tail of the exponential component. We now succeeded in preventing this problem by a new method to calculate RF without obtaining tau(e). The equation for the calculation in the new method expresses an alternans decay of PESP as a recurrence formula of PESP. It can calculate RF directly from the contractilities of the 1st through the 4th PES beats without any fitting procedure. To evaluate the reliability of the new method, we calculated RF from the alternans decay of PESP of the left ventricle (LV) of the canine excised cross-circulated heart preparation by both the original fitting and the new method. Although there was no significant difference in the mean value of the obtained RF between these two methods, the variance of RF was smaller with the new method than with the original method. Thus the new method proved useful and more reliable than the original fitting method. PMID- 11405907 TI - Acute changes in bronchoconstriction influences exhaled nitric oxide level. AB - In previous studies the exhaled nitric oxide (NO) level of asthma patients was investigated only in association with bronchial inflammation, and whether the degree of bronchoconstriction itself influences the exhaled NO level has never been investigated. We therefore evaluated the effect of inhalation of a bronchoconstrictor (methacholine) or a bronchodilator (salbutamol) on the exhaled NO level of healthy volunteers and asthma patients. The exhaled NO level of the healthy volunteers decreased after methacholine inhalation. The exhaled NO level of patients with mild or moderate persistent asthma, who had no asthma attacks on the day of measurement, increased after salbutamol inhalation, and the exhaled NO level of asthma patients during asthma attacks increased after salbutamol inhalation followed by intravenous drip infusion of aminophylline. It is suspected that large amounts of NO are trapped in the lung distal to the constricted airway, contributing little to the exhaled NO level at the mouth. However, we expect that the trapped NO is exhaled at a larger fraction after the dilatation of the constricted small airway, thereby increasing the exhaled NO level at the mouth. In conclusion, the results of this study suggest that acute changes in bronchoconstriction themselves influence the exhaled NO level independently of the change in NO synthase activity associated with airway inflammation. PMID- 11405908 TI - Evolution of the periodicity and the self-similarity in DNA sequence: a Fourier transform analysis. AB - Fourier transform analysis was applied to elucidate the periodical and self similar properties in the DNA sequences mainly of beta-globin genes in different species, and the evolutionary change in those properties was then investigated. Map patterns of a two-dimensional DNA walk showed that the stretches of exons are significantly shorter than those of introns, suggesting that the evolution of exons is driven by natural selection, whereas that of introns is generated by unknown internal rules. Using a monomer analysis, we obtained the power spectra of four different bases, A, G, C, and T, in DNA sequences. Periodicities in the short- (2 to 10 base pairs [bp]), medium- (10 to 50 bp) and long-range order (50 to 300 bp) of beta-globin gene sequences could be observed, and power spectral densities of these periodicities were increased with evolution. These results suggest the existence of the internal rules in the occurrence of the synonymous and nonsynonymous substitutions in the sequences, the destabilization of the interaction between DNA and histone protein, and the stabilization of the chromatin structure, respectively. Moreover, 1/f(alpha) analysis of the power spectra (log-log plot) in the far long-range region (160 to 16,000 bp) suggested the increase in the self-similarity (the fractal structure) of DNA sequences with evolution. A general trend of the increase in a 3 bp periodicity with evolution might be functionally related to the CAG trinucleotide repeat diseases such as Huntington chorea, where a marked periodicity of 3 bp could be observed. Fourier transform analysis applied to a DNA sequence offers a great new avenue for extracting information on the evolution of a DNA sequence. PMID- 11405909 TI - Effect of extracellular [Ca(2+)] on Ca(2+) release from sarcoplasmic reticulum in rat ventricular myocytes. AB - The effect of external [Ca(2+)] ([Ca(2+)](o)) on Ca(2+) release from the sarcoplasmic reticulum (SR) was examined with rested-state twitches in rat ventricular myocytes. The magnitude of transient rise of intracellular [Ca(2+)] ([Ca(2+)](i)) relative to the resting one, F/F(o), as measured with fluo-3, was 1.75+/-0.07 (mean+/-SEM, n=9) and 1.86+/-0.13 (n=9) at 0.3 and 1.8 mM [Ca(2+)](o), respectively; the difference was insignificant. The time from onset to peak and the rate of rise of the [Ca(2+)](i) transient were 0.107+/-0.017 s (n=9) and 18.8+/-3.38 F/F(o)/s, respectively, at 0.3 mM [Ca(2+)](o), they were 0.064+/-0.005 s (n=9) and 31.1+/-0.03 F/F(o)/s, respectively, at 1.8 mM [Ca(2+)](o). The difference in the corresponding values at the two [Ca(2+)](o) was significant (t-test, p<0.05). The half decay time of the [Ca(2+)](i) transient was 0.217+/-0.016 s (n=8) at 0.3 mM [Ca(2+)](o) and was similar to the value of 0.230+/-0.022 s (n=8) at 1.8 mM [Ca(2+)](o), indicating that the rate of decrease of [Ca(2+)](i) is independent of the [Ca(2+)](o). The duration of action potential was similar at 0.3 and 1.8 mM [Ca(2+)](o) as examined with papillary muscle. The results suggest that a lowering of [Ca(2+)](o), i.e., reducing the Ca(2+) influx, slows the rate of Ca(2+) release from the SR fully loaded with Ca(2+) with little effect on the total amount of the Ca(2+) release. An instantaneous relationship between the [Ca(2+)](i) and the myocyte shortening at 0.3 and 1.8 mM [Ca(2+)](o) suggested that the time course of unloaded contraction is related not only to the magnitude, but also to the rate of rise of [Ca(2+)](i). PMID- 11405910 TI - No dependency of a new index for oxygen cost of left ventricular contractility on heart rates in the blood-perfused excised rat heart. AB - We have reported the linear relation of myocardial oxygen consumption per beat (VO(2)) and systolic pressure-volume area (PVA) in the left ventricle of the cross-circulated rat heart. The VO(2) intercept (PVA-independent VO(2)) is primarily composed of VO(2) for Ca(2+) handling in excitation-contraction coupling and basal metabolism. Recently, we proposed a new index for oxygen cost of contractility obtainable as a slope of a linear relation between PVA independent VO(2) and left ventricular contractility. This index indicates the Ca(2+) handling VO(2) per unit contractility change. However, a dependency of this index on heart rate has not yet been investigated. The aim of the present study was to investigate the dependency of oxygen cost of contractility on heart rate. This is a critical point to compare this cost under different heart rates. At first we found no differences of VO(2)-PVA relations at 240 and 300 beats/min (bpm). Therefore, after control VO(2)-PVA relation at 300 bpm, we gradually enhanced left ventricular contractility by Ca(2+) at a midrange left ventricular volume and obtained the gradually increased PVA-independent VO(2). At each contractility level, the pacing rate was alternately changed at 240 and 300 bpm. We obtained the two composite VO(2)-PVA relation lines and found no significant differences between the slopes of PVA-independent VO(2) and left ventricular contractility relations at 240 and 300 bpm. The present results indicated no dependency of oxygen cost of left ventricular contractility on heart rates within 240--300 bpm. Based on this fact, we concluded that even under the different pacing rates within 240--300 bpm, this oxygen cost is valid for assessing cardiac mechanoenergetics, especially the economy of total Ca(2+) handling in E-C coupling. PMID- 11405911 TI - Intracellular [Ca(2+)] transients in Ca(2+) wave in single rat ventricular myocyte. AB - Ca(2+) release from the sarcoplasmic reticulum (SR) in heart muscle grades depending on Ca(2+) influx in the physiological twitch; Ca(2+( wave results from regenerative Ca(2+) release from the SR. To examine if the Ca(2+) release from the SR in the Ca(2+) wave takes a duration similar to the physiological one, a transient rise of intracellular [Ca(2+)] ([Ca(2+)](i) transient) was recorded during both a propagating Ca(2+) wave and an electrically evoked twitch with single rat ventricular myocytes, using a laser scanning confocal microscope. Care was taken to record the fluo-3 fluorescence from a segmental region with little lateral movement, especially during a propagating Ca(2+) wave. During a typical Ca(2+) wave, the time-to-peak (TP) and the half-width (HD) of the averaged [Ca(2+)](i) transient were 161 and 253 ms respectively, but they were 76 and 145 ms during an electrically evoked twitch. The difference in the duration between the two types of [Ca(2+)](i) transients could not be accounted for by modification of duration of [Ca(2+)](i) transient by possible asynchronous Ca(2+) release from the SR during a Ca(2+) wave, suggesting that the regenerative Ca2+) release from the SR in the Ca2+) wave occurs more slowly than the physiological one in rat ventricular myocytes. PMID- 11405912 TI - Caspase activation in high-pressure--induced apoptosis of murine erythroleukemia cells. AB - Murine erythroleukemia cells were subjected to physicochemical stresses such as high pressure (60--110 MPa), heating (42--45 degrees C), and ultraviolet (UV) irradiation (5--25 kJ/cm(2)). After exposure to these stresses, the cells were cultured at 37 degrees C and atmospheric pressure. The number of the cells treated at 100 MPa, 45 degrees C, or 20 kJ/cm(2) remained constant or decreased at an early stage of culture. The nuclear morphology, agarose gel electrophoresis, and flow cytometry of these cells showed that they undergo apoptosis. The activity of caspase-3 was observed in cells subjected to each stress. In particular, caspase-3 was most readily activated by high pressure under our conditions. The caspase-3 activity and apoptosis exhibited a similar pressure dependence. It is important that both caspase-3 activation and apoptosis induced by high pressure were significantly suppressed by a caspase-3 inhibitor. These results suggest that high-pressure-induced apoptosis is also characterized by the activation of caspase-3, as seen with heat- and UV-induced apoptosis. PMID- 11405913 TI - Frequency dependence of local cerebral blood flow induced by somatosensory hind paw stimulation in rat under normo- and hypercapnia. AB - We measured the field potential and the changes in local cerebral blood flow (LCBF) response during somatosensory activation (evoked LCBF) in alpha-chloralose -anesthetized rats by laser-Doppler flowmetry under normocapnia (PaCO(2)=34.3+/ 3.8 mmHg) and hypercapnia (PaCO(2)=70.1+/-9.8 mmHg). Somatosensory activation was induced by electrical stimulation (0.2, 1, and 5 Hz with 1.5 mA for 5 s) of the hind paw. The neuronal activity of the somatosensory area of the hind paw was linear to the stimulus frequency, and there was no significant difference in the neuronal activity between hypercapnia and normocapnia. The baseline level of LCBF under hypercapnia was about 72.2% higher than that under normocapnia (p<0.01). The absolute response magnitude under hypercapnia was greater than that under normocapnia (p<0.05). The evoked LCBF under both conditions showed a frequency dependent increase in the 0.2 to 5 Hz range, and the difference in the absolute response magnitude at the same stimulus frequency between normocapnia and hypercapnia became large with increasing stimulus frequency (p<0.05). On the other hand, after normalization to each baseline level there was no significant difference in the response magnitude of the normalized evoked LCBF between normocapnia and hypercapnia, indicating that the normalized evoked LCBF reflects neuronal activity even when the baseline LCBF was changed by the PaCO(2) level. The peak time and termination time of LCBF response curves with respect to the graded neuronal activity at 1 and 5 Hz stimulation increased significantly under hypercapnia, compared with those under normocapnia (p<0.05), although the rise time of 0.5 s was nearly constant. In conclusion, the results suggest a synergistic effect of the combined application of graded neuronal stimuli and hypercapnia on the LCBF response. PMID- 11405914 TI - Contractile responses of the basilar artery isolated from rabbits exposed to 8 day head-down tilt. AB - In the present study, the effects of head-down tilt (HDT) on contractile responses of basilar and popliteal arteries were investigated in vitro. The arteries were isolated from rabbits exposed to 8 d of 45 degrees HDT. Isometric tension and intracellular calcium were measured in an organ bath perfused with physiological salt solution. In the HDT rabbits, contractile response to norepinephrine (NE) was attenuated in the basilar artery, but not in the popliteal artery compared with control rabbits. HDT did not change the responses to either KCl or 5-hydroxytryptamine in both arteries. In the response of basilar artery to NE, the difference in both [Ca(2+)](i) transient and Ca sensitivity between control and HDT rabbits were statistically not significant. The response of the basilar artery to phenylephrine, a selective alpha(1)-adrenergic agonist, was also attenuated by HDT. Treatment with propranolol, a beta-adrenergic blockade, did not affect the response to NE in the basilar arteries isolated from control rabbits, but it significantly enhanced the response in the basilar arteries from HDT rabbits. These results suggest that exposure to 8-d HDT decreases a contractile response of the basilar artery to NE in rabbits. The reduction of NE-induced contraction is probably attributable to both decreased Ca transient and decreased Ca sensitivity, and decreased alpha(1)-adrenoceptor activity and increased beta-adrenoceptor activity seem to be involved in the mechanism. PMID- 11405915 TI - A new model-based method of reconstructing central aortic pressure from peripheral arterial pressure. AB - We have shown in our previous study that the transfer function between central aortic pressure and tonometric radial arterial pressure can be modeled as a pure elastic lossless tube terminated with a modified Windkessel. We hypothesized, using the model-derived radial arterial flow, that central pressure could be reconstructed by adding the time-shifted forward and backward pressure components (Stergiopulos et al.: Am J Physiol 274: H1386---H1392, 1998). In eight patients (age 16--75), central micromanometric and radial arterial tonometric pressure were measured simultaneously. We imposed measured tonometric pressure to the terminal modified Windkessel to estimate radial arterial flow, with which tonometric pressure was separated into forward and backward components. These components were then appropriately time shifted, and summed to central pressure. We used average parameter values for the terminal impedance, but individualized the transmission delay. The poor correlation (r(2)) between tonometric and central pressure (0.264--0.765) was improved by both central pressure reconstruction methods (generalized transfer function: 0.887--0.974, model-based method: 0.849--0.979). The sensitivity analysis indicated that the key model parameter in reconstructing central pressure was the transmission delay. We conclude that our model-based method was capable of reconstructing central pressure as precisely as the generalized transfer function method, and also capable of individualizing the transfer function by changing the transmission delay. PMID- 11405916 TI - The involvement of calcium mobilization in the calcium-activated potassium currents activated by hyposmotic swelling in gastric antral circular myocytes of the guinea-pig. AB - In our study of the effects of hyposmotic swelling on the Ca(2+)-activated potassium currents [I(K(Ca))] and its mechanism, we employed the whole-cell patch clamp technique using the gastric antral circular myocytes of the guinea-pig. Hyposmotic swelling efficiently increased I(K(Ca)), and the extent of changes in I(K(Ca)) was sharply dependent on the osmolarity of the perfusion solutions. When the calcium-free solution (EGTA 10 microM added in calcium-free solution) was superfused, I(K(Ca)) was not increased by the hyposmotic swelling. Gadolinium (Gd(3+)) 100 nM, a blocker of the stretch-activated nonselective cation channel, blocked the activation of I(K(Ca)) induced by hyposmotic swelling, but nicardipine 5 microM (the L-type calcium channel blocker) did not. Heparin 3 mg/ml, a potent inhibitor of inositol triphosphate receptor (InsP(3)R), did not inhibit the response, and caffeine 1 mM (the agonist for calcium-induced calcium release [CICR]) imitated the effect of hyposmotic swelling. Ryanodine (15 microM), markedly inhibited the effect. These results suggest that hyposmotic swelling activates I(K(Ca)), and the activation is associated with CICR, which is triggered by extracellular calcium influx through the stretch-activated channel (SA channel). PMID- 11405917 TI - Cardiovascular beneficial effects of electroacupuncture at Neiguan (PC-6) acupoint in anesthetized open-chest dog. AB - Neiguan (PC-6) is a traditional acupoint in the bilateral forearms, overlying the median nerve trunk. Neiguan electroacupuncture (EA) has been believed to affect cardiovascular function and used in traditional Chinese medicine to improve or treat a wide range of health conditions and diseases, including angina pectoris, myocardial infarction, hypertension, and hypotension. However, few physiological studies have assessed the beneficial effects of Neiguan EA on the cardiovascular function. In the present study, we investigated its effects on the cardiovascular function in normal open-chest dogs under pentobarbital and fentanyl anesthesia. We also obtained left ventricular (LV) pressure-volume (P-V) data with a micromanometer catheter and a volumetric conductance catheter. Mean arterial pressure, end-diastolic volume, heart rate, stroke volume, cardiac output, and end-systolic pressure gradually decreased by 5 to 10% over 1.5 h without Neiguan EA. Neiguan EA at 40 Hz, however, increased these cardiovascular variables by 10 to 15%, especially end-systolic elastance (Ees) by 40% (p<0.05) over 15 to 60 min. After Neiguan EA was stopped at 1 h, these facilitated cardiovascular variables decreased below the pre-EA level. This beneficial effect of electroacupuncture may contribute to the effectiveness of the acupuncture in Chinese medicine. PMID- 11405918 TI - Thyroid hormones may influence the slow component of VO(2) in professional cyclists. AB - We analyzed the relationship between the plasma concentrations of several hormones (testosterone [T], follicle-stimulating [FSH] and luteinizing hormone [LH], cortisol [C], 3,5,3'-triiodothyronine [T(3)], thyroxine [T(4)], and thyrotrophin [TSH]) and the magnitude of the VO(2) slow component (Delta VO(2)) in a group of nine professional road cyclists (26+/-2 years). The resting levels of the aforementioned hormones were determined before the subjects performed a 20 min cycle ergometer test at approximately 80% of VO(2 max) (or approximately 400 W). Plasma concentrations of T(3) and T(4) were inversely correlated (p<0.05) with Delta VO(2) (r=-0.72 and rr=-0.66, respectively), suggesting, at least partly, and association between thyroid basal function and the VO(2) slow component of euthyroid elite endurance athletes during constant-load intense exercise. PMID- 11405919 TI - Introductory remarks. PMID- 11405920 TI - Memorial to Bill Hamilton. PMID- 11405921 TI - Catastrophes after crossing species barriers. AB - Probably the most tragic examples of virus infections that have caused the deaths of many millions of people in the past century were the influenza and AIDS pandemics. These events occurred as a direct result of the introduction of animal viruses into the human population. Similarly, mass mortalities among aquatic and terrestrial mammals were caused by the introduction of viruses into species in which they had not previously been present. It seems paradoxical that at a time when we have managed to control or even eradicate major human virus infections like polio and smallpox we are increasingly confronted with new or newly emerging virus infections of humans and animals. A complex mix of social, technological and ecological changes, and the ability of certain viruses to adapt rapidly to a changing environment, seems to be at the basis of this phenomenon. Extensive diagnostic and surveillance networks, as well as novel vaccine- and antiviral development strategies should provide us with the safeguards to limit its impact. PMID- 11405922 TI - Epidemiology and the emergence of human immunodeficiency virus and acquired immune deficiency syndrome. AB - Although acquired immune deficiency syndrome (AIDS) was first described in the USA in 1981, there is evidence that individual cases occurred considerably earlier in Central Africa, and serological and virological data show human immunodeficiency virus (HIV) was present in the Democratic Republic of Congo (DRC) as far back as 1959. It is likely that HIV-1 infection in humans was established from cross-species transmission of simian immunodeficiency virus of chimpanzees, but the circumstances surrounding this zoonotic transfer are uncertain. This presentation will review how causality is established in epidemiology, and review the evidence (a putative ecological association) surrounding the hypothesis that early HIV-1 infections were associated with trials of oral polio vaccine (OPV) in the DRC. From an epidemiological standpoint, the OPV hypothesis is not supported by data and the ecological association proposed between OPV use and early HIV/AIDS cases is unconvincing. It is likely that Africa will continue to dominate global HIV and AIDS epidemiology in the near to medium-term future, and that the epidemic will evolve over many decades unless a preventive vaccine becomes widely available. PMID- 11405923 TI - Update on HIV/SIV infections in Cameroon. AB - The high degree of human immunodeficiency virus type 1 (HIV-1) diversity in the Cameroonian population indicates a relatively old epidemic in this country. However, studies of pygmy 'hunter-gatherers' show only rare HIV-1 infection, mainly after contact with Bantus rather than from contact with non-human primates. PMID- 11405924 TI - Experimental oral polio vaccines and acquired immune deficiency syndrome. AB - The simian immunodeficiency virus (SIV) of the common chimpanzee is widely acknowledged as the direct ancestor of HIV-1. There is increasing historical evidence that during the late 1950s, kidneys were routinely excised from central African chimpanzees by scientists who were collaborating with the polio vaccine research of Dr Hilary Koprowski, and sent - inter alia - to vaccine-making laboratories in the USA and Africa, and to unspecified destinations in Belgium. While there is no direct evidence that cells from these kidneys were used as a substrate for growing Dr Koprowski's oral polio vaccines, there is a startling coincidence between places in Africa where his CHAT vaccine was fed, and the first appearances in the world of HIV-1 group M and group-M-related AIDS. Because of the enormous implications of the hypothesis that AIDS may be an unintended iatrogenic (physician-caused) disease, it is almost inevitable that this theory will engender heated opposition from many of those in the scientific establishment, and those with vested interests. PMID- 11405925 TI - Untruths and consequences: the false hypothesis linking CHAT type 1 polio vaccination to the origin of human immunodeficiency virus. AB - A book published in 1999 hypothesized that the scientists who worked with the CHAT type 1 attenuated polio strain tested in the former Belgian Congo in the late 1950s had covertly prepared the vaccine in chimpanzee kidney cells contaminated with a simian immunodeficiency virus, which evolved into HIV-1 group M. This paper summarizes the results of the investigation conducted by the author to determine the legitimacy of the accusation. Testimony by eyewitnesses, documents of the time, epidemiological analysis, and ancillary phylogenetic, virologic and PCR data all concur to reject the hypothesis as false and without factual foundation. PMID- 11405926 TI - Postscript relating to new allegations made by Edward Hooper at The Royal Society Discussion Meeting on 11 September 2000. AB - At The Royal Society Discussion Meeting, Origins of HIV and the AIDS epidemic, which this issue records, Edward Hooper added two new 'smoking guns' to the accusations published previously in The river. These were proposed as conclusive evidence for the hypothesis that simian immunodeficiency virus-contaminated CHAT polio vaccine caused the HIV-1 group M epidemic. We have investigated the facts in relation to these 'smoking guns'. PMID- 11405927 TI - Hypotheses and facts. AB - The book, The river, is based on assumptions and not facts. Oral polio vaccine was produced entirely in rhesus monkey kidney cell cultures. Allegations that it was produced in chimpanzee kidneys at the Wistar Institute in Philadelphia or, alternatively, that the vaccine was made in the then Belgian Congo in chimpanzee kidney has no basis in fact. As the only witness to the historical events leading to the development of oral polio vaccine, I have demonstrated in this paper the truthful facts excluding any link between oral polio vaccine and human immunodeficiency virus. PMID- 11405928 TI - The Jezierski papers: live polio vaccine development in colobus monkey cells but not chimpanzee cells in the Belgian Congo, 1952-1958. AB - A reading of ten relevant papers by Alexandre Jezierski provides evidence for the only attempt in Central Africa to develop a live oral polio vaccine (OPV) from growing reference wild polio strains to 210 passages in colobus monkey tissue culture, and experimental administration to about 25 humans. Chimpanzees were used as a human model, but their tissues or kidneys were absent from the passage and production line of the proposed vaccine. Thus, the implication published by Hooper that Jezierski had produced a candidate OPV that might have contained chimpanzee viruses, possibly simian immunodeficiency virus cpz or the precursor of human immunodeficiency virus-1 group M, is incorrect. PMID- 11405929 TI - Vaccine could not have been prepared in Stanleyville. AB - In Stanleyville, at the time of vaccination campaigns, tissue cultures were primitive, experimental and used solely for diagnostic purposes. Production of vaccine was impossible to carry out. A few chimpanzee kidneys were minced and sent to Philadelphia as part of the hepatitis experiments of Dr Deinhardt. Vaccine was never handled in my laboratory and contamination with chimpanzee cells was not possible. PMID- 11405930 TI - Polio vaccine and retroviruses. AB - In this paper we consider the main steps in the process of manufacture of oral polio vaccine and assess the probable clearance factor for HIV retrovirus at each step. We conclude that the processes employed would have eliminated retrovirus contamination for all practical purposes. PMID- 11405931 TI - Simian immunodeficiency virus in kidney cell cultures from highly infected rhesus macaques (Macaca mulatta). AB - Trace amounts of simian immunodeficiency virus (SIV) proviral DNA were detected in monolayers of primary kidney cells from two rhesus macaques (Macaca mulatta) heavily infected with the highly pathogenic strain SIVmac251. There was no detectable infectious SIV in the supernatant from the kidney cell cultures obtained from either monkey. However, infectious SIV was rescued by co-culture of kidney cells with a permissive lymphoid cell line. Macrophages, present in these cultures, may be the reservoir for the proviral genomes. PMID- 11405932 TI - Responsibility for truth in research. AB - For over half a century, cell cultures derived from animals and humans have served researchers in various fields. To this day, cross-contamination of cultures has plagued many researchers, often leading to mistaken results, retractions of results, cover-ups and some out-and-out falsification of data and results following inadvertent use of the wrong cells. Also, during years of examining cultures for purity we learned that many virologists were not too concerned about the specificity of the cultures they used to propagate the particular virus under study as long as the substrate (whatever it might have been) gave optimal virus yield. Polio virus propagates in primate cells, and much research has involved cells from man and various species of primates. In the 1950s a large number of chimpanzees were held in captivity in Africa for extensive studies of the efficacy of polio vaccine in production at the Wistar Institute in Philadelphia and elsewhere. Chimpanzee tissues, particularly kidneys, were thus readily available and could have also provided substrates for polio virus production, since little was known about the purity of substrates and little attention was paid to their specificity at that time. PMID- 11405933 TI - Using human immunodeficiency virus type 1 sequences to infer historical features of the acquired immune deficiency syndrome epidemic and human immunodeficiency virus evolution. AB - In earlier work, human immunodeficiency virus type 1 (HIV-1) sequences were analysed to estimate the timing of the ancestral sequence of the main group of HIV-1, the virus that is responsible for the acquired immune deficiency syndrome pandemic, yielding a best estimate of 1931 (95% confidence interval of 1915 1941). That work will be briefly reviewed, outlining how phylogenetic tools were extended to incorporate improved evolutionary models, how the molecular clock model was adapted to incorporate variable periods of latency, and how the approach was validated by correctly estimating the timing of two historically documented dates. The advantages, limitations, and assumptions of the approach will be summarized, with particular consideration of the implications of branch length uncertainty and recombination. We have recently undertaken new phylogenetic analysis of an extremely diverse set of human immunodeficiency virus envelope sequences from the Democratic Republic of the Congo (the DRC, formerly Zaire). This analysis both corroborates and extends the conclusions of our original study. Coalescent methods were used to infer the demographic history of the HIV-1 epidemic in the DRC, and the results suggest an increase in the exponential growth rate of the infected population through time. PMID- 11405934 TI - The origins of acquired immune deficiency syndrome viruses: where and when? AB - In the absence of direct epidemiological evidence, molecular evolutionary studies of primate lentiviruses provide the most definitive information about the origins of human immunodeficiency virus (HIV)-1 and HIV-2. Related lentiviruses have been found infecting numerous species of primates in sub-Saharan Africa. The only species naturally infected with viruses closely related to HIV-2 is the sooty mangabey (Cercocebus atys) from western Africa, the region where HIV-2 is known to be endemic. Similarly, the only viruses very closely related to HIV-1 have been isolated from chimpanzees (Pan troglodytes), and in particular those from western equatorial Africa, again coinciding with the region that appears to be the hearth of the HIV-1 pandemic. HIV-1 and HIV-2 have each arisen several times: in the case of HIV-1, the three groups (M, N and O) are the result of independent cross-species transmission events. Consistent with the phylogenetic position of a 'fossil' virus from 1959, molecular clock analyses using realistic models of HIV 1 sequence evolution place the last common ancestor of the M group prior to 1940, and several lines of evidence indicate that the jump from chimpanzees to humans occurred before then. Both the inferred geographical origin of HIV-1 and the timing of the cross-species transmission are inconsistent with the suggestion that oral polio vaccines, putatively contaminated with viruses from chimpanzees in eastern equatorial Africa in the late 1950s, could be responsible for the origin of acquired immune deficiency syndrome. PMID- 11405935 TI - The origin of acquired immune deficiency syndrome: Darwinian or Lamarckian? AB - The subtypes of human immunodeficiency virus type 1 (HIV-1) group M exhibit a remarkable similarity in their between-subtype distances, which we refer to as high synchrony. The shape of the phylogenetic tree of these subtypes is referred to as a sunburst to distinguish it from a simple star phylogeny. Neither a sunburst pattern nor a comparable degree of symmetry is seen in a natural process such as in feline immunodeficiency virus evolution. We therefore have undertaken forward-process simulation studies employing coalescent theory to investigate whether such highly synchronized subtypes could be readily produced by natural Darwinian evolution. The forward model includes both classical (macro) and molecular (micro) epidemiological components. HIV-1 group M subtype synchrony is quantified using the standard deviation of the between-subtype distances and the average of the within-subtype distances. Highly synchronized subtypes and a sunburst phylogeny are not observed in our simulated data, leading to the conclusion that a quasi-Lamarckian, punctuated event occurred. The natural transfer theory for the origin of human acquired immune deficiency syndrome (AIDS) cannot easily be reconciled with these findings and it is as if a recent non-Darwinian process took place coincident with the rise of AIDS in Africa. PMID- 11405936 TI - Gene flow in wild chimpanzee populations: what genetic data tell us about chimpanzee movement over space and time. AB - The isolation of phylogenetically distinct primate immunodeficiency viruses from at least seven wild-born, captive chimpanzees indicates that viruses closely related to HIV-1 may be endemic in some wild chimpanzee populations. The search for the chimpanzee population or populations harbouring these viruses is therefore on. This paper attempts to answer the question of whether or not such populations of chimpanzees are likely to exist at all, and, if so, where they are likely to be found. We summarize what is known about gene flow in wild populations of chimpanzees, both between major phylogeographical subdivisions of the species, and within these subdivisions. Our analysis indicates that hitherto undocumented reproductively isolated chimpanzee populations may in fact exist. This conclusion is based on the observation that, despite limited geographical sampling and limited numbers of genetic loci, conventional notions of the nature and extent of chimpanzee gene flow have recently been substantially revised. Molecular genetic studies using mitochondrial DNA sequences and hypervariable nuclear microsatellite markers have indicated the existence of heretofore undocumented barriers to chimpanzee gene flow. These studies have identified at least one population of chimpanzees genetically distinct enough to be classified into a new subspecies (Pan troglodytes vellerosus). At the same time, they have called into question the long-accepted genetic distinction between eastern chimpanzees (Pan troglodytes schweinfurthii) and western equatorial chimpanzees (Pan troglodytes troglodytes). The same studies have further indicated that gene flow between local populations is more extensive than was previously thought, and follows patterns sometimes inconsistent with those documented through direct behavioural observation. Given the apparently incomplete nature of the current understanding of chimpanzee gene flow in equatorial Africa, it seems reasonable to speculate that a chimpanzee population or populations may exist which both harbour the putative HIV-1 ancestor, and which have remained reproductively isolated from other chimpanzee populations over the time-scale relevant to the evolution of the SIVcpz-HIV-1 complex of viruses. Continued extensive sampling of wild chimpanzee populations, both for their genes and their viruses, should be performed quickly considering the high probability of extinction that many wild chimpanzee populations face today. The history of human-chimpanzee contacts is discussed. PMID- 11405937 TI - Infectious disease dynamics: What characterizes a successful invader? AB - Against the background of human immunodeficiency virus (HIV) and acquired immune deficiency syndrome (AIDS) and other potentially emerging (or re-emerging) infectious diseases, this review will focus on the properties which enable an infectious agent to establish and maintain itself within a specified host population. We shall emphasize that for a pathogen to cross a species barrier is one thing, but for it successfully to maintain itself in the new population is must have a 'basic reproductive number', R(0), which satisfies R(0) > 1. We shall further discuss how behavioural factors interweave with the basic biology of the production of transmission stages by the pathogen, all subject to possible secular changes, to determine the magnitude of R(0). Although primarily focusing on HIV and AIDS, we shall review wider aspects of these questions. PMID- 11405938 TI - Serial human passage of simian immunodeficiency virus by unsterile injections and the emergence of epidemic human immunodeficiency virus in Africa. AB - There is compelling evidence that both human immunodeficiency virus (HIV) types emerged from two dissimilar simian immunodeficiency viruses (SIVs) in separate geographical regions of Africa. Each of the two HIVs has its own simian progenitor and specific genetic precursor, and all of the primates that carry these SIVs have been in close contact with humans for thousands of years without the emergence of epidemic HIV. To date no plausible mechanism has been identified to account for the sudden emergence in the mid-20th century of these epidemic HIVs. In this study we examine the conditions needed for SIV to complete the genetic transition from individual human SIV infections to epidemic HIV in humans. The genetic distance from SIV to HIV and the mutational activity needed to achieve this degree of adaptation to human hosts is placed within a mathematical model to estimate the probabilities of SIV completing this transition within a single SIV-infected human host. We found that the emergence of even one epidemic HIV strain, following a single human exposure to SIV, was very unlikely. And the probability of four or more such transitions (i.e. HIV-1 groups M, O and HIV-2 subtypes A and B) occurring in a brief period is vanishingly small. We conclude that SIV cannot become a zoonosis, but requires adaptive mutations to become HIV. Some modern event must have aided in the transition of SIV to HIV. Our research indicates that serial passage of partially adapted SIV between humans could produce the series of cumulative mutations sufficient for the emergence of epidemic HIV strains. We examined the rapid growth of unsterile injections in Africa beginning in the 1950s as a biologically plausible event capable of greatly increasing serial human passage of SIV and generating HIV by a series of multiple genetic transitions. We conclude that increased unsterile injecting in Africa during the period 1950-1970 provided the agent for SIV human infections to emerge as epidemic HIV in the modern era. PMID- 11405939 TI - Man, monkeys and malaria. AB - Bizarre though it may now seem, in the last century a whole series of experiments was conducted that involved injecting fresh monkey blood into human volunteers or patients. The reasons, valid at the time, were either to treat neurosyphilis with a relatively benign simian malaria infection (so-called pyrogen therapy), or to establish which monkey malaria species were potential zoonotic reservoirs of infection that then may have interfered with malaria eradication campaigns. Although direct inoculation of fresh blood is the most effective way of retroviruses as well as malaria parasites crossing the species barrier, this hypothesis was never taken up or researched. Unlikely, but not disproved, it is important to remember some of the more hazardous experiments that were done in good faith, too long ago to be recorded on electronic databases. PMID- 11405940 TI - The earliest cases of human immunodeficiency virus type 1 group M in Congo Kinshasa, Rwanda and Burundi and the origin of acquired immune deficiency syndrome. AB - The early cases of acquired immune deficiency syndrome and human immunodeficiency virus type 1 (HIV-1) infection in the 1960s and 1970s in Congo-Kinshasa (Zaire), Rwanda and Burundi are reviewed. These countries appear to be the source of the HIV-1 group M epidemic, which then spread outwards to neighbouring Tanzania and Uganda in the east, and Congo-Brazzaville in the west. Further spread to Haiti and onwards to the USA can be explained by the hundreds of single men from Haiti who participated in the UNESCO educational programme in the Congo between 1960 and 1975. PMID- 11405941 TI - The distribution of early acquired immune deficiency syndrome cases and conditions for the establishment of new epidemics. AB - This article presents discussion on two issues: the distribution of early cases of acquired immune deficiency syndrome (AIDS), and the epidemiological conditions for the establishment of new diseases. Evidence from four presentations on early AIDS cases is discussed, together with issues of interpreting association and inferring causation. In the second section, the results of a stochastic epidemiological model using the 'real' geography of villages in northern Zaire is presented. This is used to investigate the conditions under which newly introduced infections with different characteristics are able to establish in human populations. It highlights additional spatial, temporal and behavioural conditions necessary for the persistence of introduced diseases, in addition to the condition that the basic reproductive rate R(0) > 1. PMID- 11405942 TI - Concerning the early and current spread of human immunodeficiency virus type 1 and the scientific burden of proof versus the burden of current human immunodeficiency virus incidence. AB - The following points were made during the Discussion that followed the paper by Brian Martin. PMID- 11405943 TI - The origin of acquired immune deficiency syndrome: Can science afford to ignore it? AB - There is a crisis of public faith in science and scientists. Recent research shows concern over scientific ethics, transparency and who benefits from research and development, exemplified in the genetically modified organism debate. Scientific discussion of the polio vaccine hypothesis for the origin of acquired immune deficiency syndrome (AIDS) has been systematically suppressed for more than 12 years. The author calls for an international multidisciplinary inquiry into the origin of AIDS, arguing it is essential to human health, prevention of new pandemics, and to protect the integrity of science in the eyes of the public. PMID- 11405944 TI - The burden of proof and the origin of acquired immune deficiency syndrome. AB - There is a distinct difference in the way that different theories about the origin of acquired immune deficiency syndrome have been treated, with the widely supported cut-hunter theory given relatively little scrutiny, while the oral polio vaccine theory has been subject to intense criticism. This difference in treatment cannot be explained as application of the scientific method. A better explanation is that the burden of proof is put on all contenders to the cut hunter theory, giving it an unfair advantage, especially given that this assignment of the burden of proof appears to reflect non-scientific factors. PMID- 11405945 TI - Natural and iatrogenic factors in human immunodeficiency virus transmission. AB - In the light of the evidence and discussion presented during The Royal Society Discussion Meeting it seems to me that the oral polio vaccine (OPV) hypothesis for the origins of human immunodeficiency virus (HIV) and the acquired immune deficiency syndrome epidemic is less tenable now than one year earlier. The OPV hypothesis does not accord with HIV phylogenetic studies: the geographical correlation has been challenged; the testimony of those directly involved with OPV trial vaccines denies the use of chimpanzees, corroborating tests on the still-available vials of the CHAT vaccines, which contain neither simian immunodeficiency virus nor chimpanzee DNA. Yet one lesson to be learned from considering OPV as a source of HIV is how plausibly it might have happened and how cautious we need to be over introducing medical treatments derived from animal tissues, such as live, attenuated vaccines or xenotransplantation. To cast doubt on the OPV hypothesis is not to dismiss entirely the role of iatrogenic factors in HIV transmission from chimpanzees in the first instance, in HIV adaptation to onward transmission during its early phase in humans, or in the later spread of HIV to patients, for example, with haemophilia. To reduce the argument over the origins of HIV to the 'OPV hypothesis' versus the 'cut-hunter hypothesis' is an oversimplistic and false antithesis. Both natural and iatrogenic transmission of many retroviruses, including HIV, have been thoroughly documented and are not mutually exclusive. Exactly how, when and where the first human(s) became infected with the progenitor of HIV-1 group M, which gave rise to the pandemic strain, is likely, however, to remain a matter of conjecture. PMID- 11405946 TI - The Leeuwenhoek Lecture 2001. Animal origins of human infectious disease. AB - Since time immemorial animals have been a major source of human infectious disease. Certain infections like rabies are recognized as zoonoses caused in each case by direct animal-to-human transmission. Others like measles became independently sustained with the human population so that the causative virus has diverged from its animal progenitor. Recent examples of direct zoonoses are variant Creutzfeldt-Jakob disease arising from bovine spongiform encephalopathy, and the H5N1 avian influenza outbreak in Hong Kong. Epidemics of recent animal origin are the 1918-1919 influenza pandemic, and acquired immune deficiency syndrome caused by human immunodeficiency virus (HIV). Some retroviruses jump into and out of the chromosomal DNA of the host germline, so that they oscillate between being inherited Mendelian traits or infectious agents in different species. Will new procedures like animal-to-human transplants unleash further infections? Do microbes become more virulent upon cross-species transfer? Are animal microbes a threat as biological weapons? Will the vast reservoir of immunodeficient hosts due to the HIV pandemic provide conditions permissive for sporadic zoonoses to take off as human-to-human transmissible diseases? Do human infections now pose a threat to endangered primates? These questions are addressed in this lecture. PMID- 11405947 TI - Antipsychotic medication for those with both schizophrenia and learning disability. AB - BACKGROUND: Antipsychotic medication is the standard treatment for people with learning disability and schizophrenia. OBJECTIVES: To determine the efficacy of any antipsychotic medication for treating people with a dual diagnosis of learning disability and schizophrenia. SEARCH STRATEGY: Electronic searching of Biological Abstracts, the Cochrane Schizophrenia Group's Register of trials (September 2000), the Cochrane Library (Issue 3 2000), EMBASE, PsycLIT MEDLINE and National Research Register. Unpublished data were sought from pharmaceutical companies. Both authors independently selected the relevant studies from the reports identified in this way. SELECTION CRITERIA: 1. All randomised controlled trials of antipsychotic medication, regardless of dosage, versus a placebo control, of longer than one month's duration. 2. Anyone over 18 years of age with both learning disability and schizophrenia. Learning disability was defined as a measured IQ of 70 or less. Any mode of diagnosis of schizophrenia was acceptable. DATA COLLECTION AND ANALYSIS: The two reviewers independently attempted to select and then extract data but it was not possible to do this with the single study that met the inclusion criteria. MAIN RESULTS: Only one relevant randomised trial was found by the searches. This study included four people with a dual diagnosis of schizophrenia and learning disability, but results were available for only two. The groups to which the other two people were allocated were unclear. In order to display the data, too many assumptions would have to have been made about these other two people and any results would be uninformative and potentially misleading. REVIEWER'S CONCLUSIONS: Using the methods described the reviewers found no randomised controlled trial evidence to guide the use of antipsychotic medication for those with both learning disability and schizophrenia. Until the urgent need for randomised controlled trials is met, clinical practice will continue to be guided by extrapolation of evidence from randomised controlled trials involving people with schizophrenia but without learning disability and non-randomised trials of those with learning disability and schizophrenia. PMID- 11405948 TI - Treatments for breast engorgement during lactation. AB - BACKGROUND: National surveys have shown that painful breasts are the second most common reason for giving up breastfeeding in the first two weeks after birth in the UK. One factor contributing to such pain can be breast engorgement. Views differ as to how engorgement arises, although restrictive feeding patterns in hospital are likely to have contributed in the past. These differing views are reflected in the range of solutions offered to treat engorgement in breastfeeding mothers and these treatments are assessed in this review. OBJECTIVES: To determine the effects of any proposed intervention to relieve symptoms of breast engorgement among breastfeeding women. SEARCH STRATEGY: The register of clinical trials maintained and updated by the Cochrane Pregnancy and Childbirth Group. CINAHL and MEDLINE were also searched. Date of last search: December 2000. SELECTION CRITERIA: All randomised and 'quasi-randomised' controlled trials, with or without blinding, that assess the effectiveness of treatments for the alleviation of symptoms in breastfeeding women experiencing engorgement. DATA COLLECTION AND ANALYSIS: Data were extracted by one reviewer and verified by a second reviewer. MAIN RESULTS: Eight trials, involving 424 women, were included. Three different studies were identified which used cabbage leaves or cabbage leaf extracts;. no overall benefit was found. Ultrasound treatment and placebo were equally effective. Use of Danzen (an anti-inflammatory agent) significantly improved the total symptoms of engorgement when compared to placebo (odds ratio (OR) 3.6, 95% confidence interval (CI) 1.3 - 10.3) as did bromelain/trypsin complex (OR 8.02, 95% CI 2.8-23.3). Oxytocin and cold packs had no demonstrable effect on engorgement symptoms. REVIEWER'S CONCLUSIONS: Cabbage leaves and gel packs were equally effective in the treatment of engorgement. Since both cabbage extract and placebo cream were equally effective, the alleviation in symptoms may be brought about by other factors, such as breast massage. Ultrasound treatment is equally effective with or without the ultra-wave emitting crystal, therefore its effectiveness is more likely to be due to the effect of radiant heat or massage. Pharmacologically, oxytocin was not an effective engorgement treatment while Danzen and bromelain/trypsin complex significantly improved the symptoms of engorgement. Initial prevention of breast engorgement should remain the key priority. PMID- 11405949 TI - Continuous electronic heart rate monitoring for fetal assessment during labor. AB - BACKGROUND: Electronic fetal monitoring (EFM) is used in the management of labor and delivery in nearly three of four pregnancies in the United States. The apparent contradiction between the widespread use of EFM and expert recommendations to limit routine use indicates that a reassessment of this practice is warranted. OBJECTIVES: To compare the efficacy and safety of routine continuous EFM during labor with intermittent auscultation, using the results of published randomized controlled trials (RCTs). SEARCH STRATEGY: We identified RCTs by searching MEDLINE and the register maintained by the Cochrane Pregnancy and Childbirth Group, and by contacting experts, and reviewing published references. Date of last search: January 2001. SELECTION CRITERIA: Randomized controlled trials. DATA COLLECTION AND ANALYSIS: Data were abstracted by one of us, and their accuracy was confirmed independently by a second person. A single reviewer assessed study quality based on criteria developed by others for RCTs. Data reported from similar studies were used to calculate a combined risk estimate for each of eight outcomes. MAIN RESULTS: Our search identified 13 published RCTs addressing the efficacy and safety of EFM; no unpublished studies were found. Four trials that did not fulfil our selection criteria were excluded. The remaining nine trials included 18,561 pregnant women and their 18,695 infants in both high- and low-risk pregnancies from seven clinical centers in the United States, Europe, and Australia. Overall, a statistically significant decrease was associated with routine EFM for neonatal seizures (relative risk (RR) 0.51, 95% confidence interval (CI) 0.32-0.82). The protective effect for neonatal seizures was only evident in studies with high-quality scores. No significant differences were observed in 1-minute Apgar scores below four or seven, rate of admissions to neonatal intensive care units, perinatal deaths or cerebral palsy. An increase associated with the use of EFM was observed in the rate of cesarean delivery (RR 1.41, 95% CI 1.23-1.61) and operative vaginal delivery (RR 1.20, 95% CI 1.11 1.30). REVIEWER'S CONCLUSIONS: The only clinically significant benefit from the use of routine continuous EFM was in the reduction of neonatal seizures. In view of the increase in cesarean and operative vaginal delivery, the long-term benefit of this reduction must be evaluated in the decision reached jointly by the pregnant woman and her clinician to use continuous EFM or intermittent auscultation during labor. PMID- 11405950 TI - Elective caesarean section versus expectant management for delivery of the small baby. AB - BACKGROUND: Elective caesarean delivery for women in labour with a small or immature baby might reduce the chances of fetal or neonatal death, but it might also increase the risk of maternal morbidity. OBJECTIVES: To assess the effects of a policy of elective caesarean delivery versus expectant management for small babies. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register was searched. Date of last search: December 2000. SELECTION CRITERIA: Randomised trials comparing a policy of elective caesarean delivery versus expectant management with recourse to caesarean section. DATA COLLECTION AND ANALYSIS: One reviewer assessed eligibility and trial quality, and both contributed to the update. MAIN RESULTS: Six studies involving 122 women were included. All trials reported recruiting difficulties. Babies in the elective group were less likely to have respiratory distress syndrome (odds ratio (OR) 0.43, 95% confidence interval (CI) 0.18 to 1.06) although they were more likely to have a low cord pH immediately after delivery (OR 10.82, 95% CI 1.60 to 73.24). They were less likely to have neonatal seizures (0/39 versus 2/42) and there were fewer deaths (2/62 versus 6/60) but these differences did not reach statistical significance. However, their mothers were more likely to have serious morbidity (OR 6.44, 95% CI 1.48 to 27.89). REVIEWER'S CONCLUSIONS: There is not enough evidence to evaluate the use of a policy for elective caesarean delivery for small babies. Randomised trials in this area are likely to continue to experience recruitment problems. However, it still may be possible to investigate elective caesarean delivery in small babies with cephalic presentations. PMID- 11405951 TI - Natural surfactant extract versus synthetic surfactant for neonatal respiratory distress syndrome. AB - BACKGROUND: Surfactant preparations are now widely used and have been credited with recent improvements in overall infant mortality (Horbar 1993b, Schwartz 1994). A wide variety of surfactant preparations have been developed and tested. These include synthetic surfactants and surfactants derived from animal sources. Although clinical trials have demonstrated that both synthetic surfactants and natural surfactant preparations are effective, comparison in animal models has suggested that there may be greater efficacy of natural surfactant products, perhaps due to the protein content of natural surfactant (Tooley 1987). OBJECTIVES: To compare the effect of synthetic surfactant to natural surfactant in premature infants at risk for or having respiratory distress syndrome. SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline 1975 through December 2000 (MeSH terms: pulmonary surfactant; limits: age groups, newborn infant; publication type, clinical trial), previous reviews including cross references, abstracts, conference and symposia proceedings, expert informants, and journal hand searching in the English language. SELECTION CRITERIA: Randomized controlled trials comparing administration of synthetic surfactants to administration of natural surfactant extracts in premature infants at risk for or having respiratory distress syndrome were considered for this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including pneumothorax, patent ductus arteriosus, necrotizing enterocolitis, intraventricular hemorrhage (all intraventricular hemorrhage and severe intraventricular hemorrhage), bronchopulmonary dysplasia, chronic lung disease, retinopathy of prematurity, and mortality were excerpted by both reviewers. Data analysis was conducted according to the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Eleven trials met inclusion criteria. The meta-analysis shows that the use of natural surfactant rather than synthetic surfactant results in a significant reduction in the risk of pneumothorax (typical relative risk 0.63, 95% CI 0.53, 0.75; typical risk difference -0.04, 95% CI -0.06, -0.03) and the risk of mortality (typical relative risk 0.87, 95% CI 0.76, 0.98; typical risk difference -0.02, 95% CI -0.05, 0.00). Natural surfactant extract is associated with a marginal increase in the risk of intraventricular hemorrhage (typical relative risk 1.09, 95% CI 1.00, 1.19; typical risk difference 0.03, 95% CI 0.00, 0.06), but no increase in grade 3 to 4 intraventricular hemorrhage (typical relative risk 1.08, 95% CI 0.92, 1.28; typical risk difference 0.01, 95% CI -0.01, 0.03). The meta-analyses support a marginal decrease in the risk of bronchopulmonary dysplasia or mortality associated with the use of natural surfactant preparations (typical relative risk 0.95, 95% CI 0.90, 1.01; typical risk difference -0.03, 95% CI -0.06, 0.00). No other relevant differences in outcome are noted. REVIEWER'S CONCLUSIONS: Both natural surfactant extracts and synthetic surfactant extracts are effective in the treatment and prevention of respiratory distress syndrome. Comparative trials demonstrate greater early improvement in the requirement for ventilator support, fewer pneumothoraces, and fewer deaths associated with natural surfactant extract treatment. Natural surfactant may be associated with an increase in intraventricular hemorrhage, though the more serious hemorrhages (Grade 3 and 4) are not increased. Despite these concerns, natural surfactant extracts would seem to be the more desirable choice when compared to currently available synthetic surfactants. PMID- 11405952 TI - Phenobarbital prior to preterm birth for preventing neonatal periventricular haemorrhage. AB - BACKGROUND: Preterm infants are at risk of periventricular haemorrhage. This can be a sign of brain damage that might lead to neurodevelopmental abnormalities, including cerebral palsy. Phenobarbital might prevent ischaemic injury or reduce fluctuations in blood pressure and blood flow in the brain. OBJECTIVES: The objective of this review was to assess the benefits and harms of giving phenobarbital to women at risk of imminent very preterm birth with the primary aim of preventing periventricular haemorrhage in the infant. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies. Date of last search: September 2000. SELECTION CRITERIA: Randomised trials with reported data which compare outcomes, such as neonatal mortality, neonatal neurological and other morbidity, long term neurodevelopment and maternal morbidity, following prenatal exposure to phenobarbital, with outcomes in controls with or without placebo. DATA COLLECTION AND ANALYSIS: Assessments of trial eligibility, quality and data extractions were done by the two authors independently. Eligible trials were included in the initial analysis and prespecified sensitivity analyses done to evaluate the effect of trial quality. MAIN RESULTS: Over 1600 women were entered into the eight trials included. Analyses showed a significant reduction in the rates of all grades of periventricular haemorrhage (PVH) (relative risk 0.72, 95% confidence interval 0.62 to 0.84) and severe grades PVH (3 and 4) (relative risk 0.48, 95% confidence interval 0.32 to 0.72) in infants whose mothers had been given prenatal phenobarbital. These results were influenced by the earlier trials which were of poorer quality and contributed excessive weight in the analysis due to their higher rates of severe PVH. Over time, and with improved trial quality, these beneficial effects disappeared. No difference was found in the incidence of neurodevelopmental abnormalities at paediatric follow-up assessed between 18-36 months of age. Maternal sedation was more likely in women receiving phenobarbital. REVIEWER'S CONCLUSIONS: Phenobarbital administration to women prior to preterm birth cannot be recommended for routine clinical practice. Any future trials should examine the effects of phenobarbital prior to preterm birth at gestational ages with a high risk of PVH, stratify for gestational age and ensure minimal exclusions after randomisation. Neurodevelopmental status at follow-up should be measured as the most important outcome. PMID- 11405953 TI - Physician advice for smoking cessation. AB - BACKGROUND: Health care professionals frequently advise patients to improve their health by stopping smoking. Such advice may be brief, or part of more intensive interventions. OBJECTIVES: The aims of this review were to assess the effectiveness of advice from physicians in promoting smoking cessation; to compare minimal interventions by physicians with more intensive interventions; to assess the effectiveness of various aids to advice in promoting smoking cessation and to determine the effect of anti-smoking advice on disease specific and all cause mortality. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register and the Cochrane Controlled Trials Register. Date of the most recent searches: October 2000. SELECTION CRITERIA: Randomized trials of smoking cessation advice from a medical practitioner in which abstinence was assessed at least six months after advice was first provided. DATA COLLECTION AND ANALYSIS: We extracted data in duplicate on the setting in which advice was given, type of advice given (minimal or intensive), and whether aids to advice were used, the outcome measures, method of randomization and completeness of followup. The main outcome measures were abstinence from smoking after at least six months follow-up and mortality. We used the most rigorous definition of abstinence in each trial, and biochemically validated rates where available. Subjects lost to follow-up were counted as smokers. Where possible, meta-analysis was performed using a fixed effects model. MAIN RESULTS: We identified thirty four trials, conducted between 1972 and 1999, including over 27,000 smokers. In some trials, subjects were at risk of specified diseases (chest disease, diabetes, ischaemic heart disease), but most were from unselected populations. The most common setting for delivery of advice was primary care. Other settings included hospital wards and outpatient clinics, and industrial clinics. Pooled data from 16 trials of brief advice versus no advice (or usual care) revealed a small but significant increase in the odds of quitting (odds ratio 1.69, 95% confidence interval 1.45 to 1.98). This equates to an absolute difference in the cessation rate of about 2.5%. There was insufficient evidence, from indirect comparisons, to establish a significant difference in the effectiveness of physician advice according to the intensity of the intervention, the amount of follow-up provided, and whether or not various aids were used at the time of the consultation in addition to providing advice. However, direct comparison of intensive versus minimal advice showed a small advantage of intensive advice (odds ratio 1.44, 95% confidence interval 1.23 to 1.68). Only one study determined the effect of smoking advice on mortality. It found no statistically significant differences in death rates at twenty years follow-up. REVIEWER'S CONCLUSIONS: Simple advice has a small effect on cessation rates. Additional manoeuvres appear to have only a small effect, though more intensive interventions are marginally more effective than minimal interventions. PMID- 11405954 TI - Short-term low-dose corticosteroids vs placebo and nonsteroidal antiinflammatory drugs in rheumatoid arthritis. AB - BACKGROUND: The effect of low dose corticosteroids, equivalent to 15 mg prednisolone daily or less, in patients with rheumatoid arthritis has been questioned. We therefore performed a systematic review of trials which compared corticosteroids with placebo or non-steroidal, anti-inflammatory drugs. OBJECTIVES: To determine whether short-term (i.e. as recorded within the first month of therapy), oral low-dose corticosteroids (corresponding to a maximum of 15 mg prednisolone daily) is superior to placebo and nonsteroidal, antiinflammatory drugs in patients with rheumatoid arthritis. SEARCH STRATEGY: Medline Silverplatter, The Cochrane Controlled Trials Register, reference lists and a personal archive. SELECTION CRITERIA: All randomised studies comparing an oral corticosteroid (not exceeding an equivalent of 15 mg prednisolone daily) with placebo or a nonsteroidal, antiinflammatory drug were eligible if they reported clinical outcomes within one month after start of therapy. DATA COLLECTION AND ANALYSIS: Decisions on which trials to include were made independently by two observers based on the methods sections of the trials only. Standardised effect measures were used for the statistical analyses; the random effects model was used if P<0.10 for the test of heterogeneity. MAIN RESULTS: Ten studies, involving 320 patients, were included in the meta-analysis. Prednisolone had a marked effect over placebo on joint tenderness (standardised effect size 1.31, 95% confidence interval 0.78 to 1.83), pain (standardised effect size 1.75, 0.87 to 2.64) and grip strength (standardised effect size 0.41, 0.13 to 0.69). Measured in the original units, the differences were 12 tender joints (6 to 18) and 22 mm Hg (5 to 40) for grip strength. Prednisolone also had a greater effect than nonsteroidal, antiinflammatory drugs on joint tenderness (standardised effect size 0.63, 0.11 to 1.16) and pain (standardised effect size 1.25, 0.26 to 2.24), whereas the difference in grip strength was not significant (standardised effect size 0.31, -0.02 to 0.64). Measured in the original units, the differences were 9 tender joints (5 to 12) and 12 mm Hg (-6 to 31). The risk of adverse effects, also during moderate- and long-term use, seemed acceptable. REVIEWER'S CONCLUSIONS: Prednisolone in low doses (not exceeding 15 mg daily) may be used intermittently in patients with rheumatoid arthritis, particularly if the disease cannot be controlled by other means. Since prednisolone is highly effective, short-term placebo controlled trials studying the clinical effect of low-dose prednisolone or other oral corticosteroids are no longer necessary. PMID- 11405955 TI - Gamma-aminobutyric acid agonists for neuroleptic-induced tardive dyskinesia. AB - BACKGROUND: Chronic antipsychotic drug treatment may cause tardive dyskinesia (TD), a long-term movement disorder. The gamma-aminobutyric acid (GABA) agonist drugs have been trialed as a treatment for TD, but these drugs have intense sedative properties and may exacerbate psychotic symptoms. OBJECTIVES: To determine the effects of GABA agonist drugs (baclofen, gamma-vinyl-GABA, gamma acetylenic-GABA, progabide, muscimol, sodium valproate and tetrahydroisoxazolopyridine (THIP)) for people with neuroleptic-induced tardive dyskinesia (TD) and schizophrenia or other chronic mental illnesses. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1982-2000), The Cochrane Library (Issue 4, 2000), Cochrane Schizophrenia Group's Register of Trials (2000), EMBASE (1980-2000), LILACS (1982-2000), MEDLINE (1966-2000), PsycLIT (1974-2000), and SCISEARCH were undertaken. References of all identified studies were searched for further trial citations. First authors of each included trial were contacted. SELECTION CRITERIA: The inclusion criteria for all relevant randomised studies were that they should focus on people with schizophrenia or other chronic mental illnesses, with neuroleptic-induced TD and compare the use of non-benzodiazepine GABA agonist drugs to placebo or no intervention. DATA COLLECTION AND ANALYSIS: The reviewers extracted the data independently and the relative risk (RR) and its 95% confidence interval (CI) or the weighted mean difference with 95% CI were estimated. The reviewers assumed that people who dropped out had no improvement. MAIN RESULTS: Eight small, short, poorly reported studies were included. For the outcome of 'no clinically important improvement in tardive dyskinesia' GABA agonist drugs were not clearly better than placebo (n=108, RR 0.83 CI 0.6 to 1.1). Deterioration in mental state was more likely to occur for people receiving GABA medication (n=95, RR 2.55 CI 1.17 to 5.59), but this effect was influenced by the decision to assign a negative outcome to those who dropped out before the end of the study. A greater proportion of people allocated GABA medication may fail to complete the trial compared with those allocated placebo (20% versus 9%) but, this difference was not statistically significant (n=136, RR 1.99 CI 0.84 to 4.68). There is a suggestion of an increase in ataxia for both baclofen and sodium valproate (n=95, RR 3.26 CI 0.4 to 30), and in sedation (n=113, RR 2.12 CI 0.8 to 5.4) compared with placebo. Withdrawal of THIP may cause seizures. REVIEWER'S CONCLUSIONS: Currently, evidence of effect of baclofen, progabide, sodium valproate, or THIP for people with neuroleptic-induced TD is inconclusive and unconvincing. Any possible benefits are likely to be outweighed by the adverse effects associated with the use of these drugs. PMID- 11405956 TI - Interventions for treating acute and chronic Achilles tendinitis. AB - BACKGROUND: Achilles tendinitis is one of the most common of all sports injuries. There is no consensus on treatment. OBJECTIVES: To assess the effectiveness of various treatment interventions for acute and chronic Achilles tendinitis in adults. SEARCH STRATEGY: The Cochrane Musculoskeletal Injuries Group specialised register (December 2000), Cochrane Controlled Trials Register (The Cochrane Library Issue 4, 2000), MEDLINE (1966 to December 2000), EMBASE (1980 to 2001 wk 04), CINAHL (1982 to December 2000), and reference lists of identified trials were searched. SELECTION CRITERIA: Randomised or quasi-randomised trials of treatment interventions for acute and chronic Achilles tendinitis in adults. Studies focusing on pathological tendinitis were excluded. Excluded were those trials that compared different dosages of the same drug or drugs within the same class of drugs, for example different non-steroidal anti-inflammatory drugs (NSAIDs). DATA COLLECTION AND ANALYSIS: Three reviewers independently assessed trial quality, by use of a ten item check list, and extracted data. Requests were sent for separate data for Achilles tendinitis patients in studies within trials of mixed patient populations. Where possible, quantitative analysis and limited pooling of data were undertaken. MAIN RESULTS: Nine trials, involving 697 patients, met the inclusion criteria of the review. Methodological quality was adequate in most of the trials with regards to blinding but the assessment of outcome was incomplete and short-term. There was weak but not robust evidence from three trials of a modest benefit of NSAIDs for the alleviation of acute symptoms. There was some weak evidence of no difference compared with no treatment of low dose heparin, heel pads, topical laser therapy and peritendonous steroid injection, but this could not be fully evaluated from the reports of four trials. The results of an experimental preparation of a calf-derived deproteinized haemodialysate, Actovegin, were promising but the severity of patient symptoms was questionable in the single small trial testing this comparison. The results of a comparison of glycosaminoglycan sulfate with a NSAID were inconclusive. REVIEWER'S CONCLUSIONS: There is insufficient evidence from randomised controlled trials to determine which method of treatment is the most appropriate for the treatment of acute or chronic Achilles tendinitis. Further research is warranted. PMID- 11405957 TI - Compression for venous leg ulcers. AB - OBJECTIVES: To assess the effectiveness and cost-effectiveness of compression bandaging and stockings in the treatment of venous leg ulcers. SEARCH STRATEGY: Searches of 19 databases, hand searching of journals, conference proceedings and bibliographies. Manufacturers of compression bandages and stockings and an Advisory Panel were contacted for unpublished studies. SELECTION CRITERIA: Trials that evaluated compression bandaging or stockings, as a treatment for venous leg ulcers. There was no restriction on date or language. Ulcer healing was the primary endpoint. DATA COLLECTION AND ANALYSIS: Details of eligible studies were extracted and summarised using a data extraction sheet. Data extraction was verified by two reviewers independently. MAIN RESULTS: Twenty two trials reporting 24 comparisons were identified. Compression was more effective than no compression (4/6 trials). When multi-layered systems were compared, elastic compression was more effective than non-elastic compression (5 trials). There was no difference in healing rates between 4-layer bandaging and other high compression multi-layered systems (3 trials). There was no difference in healing rates between elastomeric multi-layered systems (4 trials). Multi-layered high compression was more effective than single layer compression (4 trials). Compression stockings were evaluated in two trials. One found a high compression stocking plus a thrombo stocking to be more effective than a short stretch bandage. The second small trial reported no difference between the compression stockings and Unna's boot. There were insufficient data to draw conclusions about the relative cost-effectiveness of different regimens. REVIEWER'S CONCLUSIONS: Compression increases ulcer healing rates compared with no compression. Multi layered systems are more effective than single-layered systems. High compression is more effective than low compression but there are no clear differences in the effectiveness of different types of high compression. PMID- 11405958 TI - Dehydroepiandrosterone (DHEA) supplementation for cognitive function . AB - BACKGROUND: In view of the theoretical rationale for beneficial effects of DHEA and DHEAS on cognitive function in ageing and dementia, we have undertaken a thorough investigation of well-conducted studies in this area. This will provide a basis for confirmation of any effect of DHEA/S administration in humans in properly controlled trials. The review will also provide a scientific basis for effective dosage, acceptable route and duration of administration, and side effect profiles. This review is especially pertinent at this time as DHEA is currently being sold in large quantities in health food stores, particularly in the USA. In some cases the recommended dose is different for men and women (50mg/day for men and 25mg/day for women) and the basis for this recommendation needs to be explored. OBJECTIVES: To establish whether administration of DHEA, or its sulphate, DHEAS, improves cognitive function or reduces the rate of decline of cognitive function in normal older adults or in individuals with dementia. SEARCH STRATEGY: Relevant electronic databases, journals, personal communications and conference abstracts were searched for randomised controlled trials investigating the effects of DHEA/S on cognition in older adults. SELECTION CRITERIA: All relevant randomised controlled trials of DHEA/S were considered for inclusion in the review. DATA COLLECTION AND ANALYSIS: Data for the specified outcomes were independently extracted by two reviewers (FAH & JvN) and cross checked. Any discrepancies were discussed and resolved. No data pooling was undertaken owing to the lack of availability of the relevant statistics. MAIN RESULTS: There are four included studies, three cognition in normal older people, and Barnhart 1999 in perimenopausal women with decreased well-being. There were no studies in dementia. There were a few significant findings. Wolf 1997 found significant improvement following DHEA compared with placebo in both immediate recall (MD 0.8, 95% CI 0.16, 1.44) and delayed recall (MD 0.9, 95% CI 0.09, 1.71) of a visual memory test in women, estimated in a crossover trial after 2 weeks of treatment with each of DHEA and placebo. However there was no significant improvement in men, nor a significant effect on a verbal memory test. There was also no significant effect on four other cognitive tests. Wolf 1998 (2) found that placebo group performance deteriorated significantly on a test of selective attention following a psychosocial stressor (p<0.05), while deterioration was not evident in the DHEA group (p=0.85) after two weeks of treatment. However, when compared to placebo, DHEA produced a significant impairment on a visual memory test (p<0.01) following the stressor. No significant effect was found on a third cognitive task. Effects were not found on tasks when administered in the absence of a stressor. Barnhart 1999 employed three cognitive measures and found no significant effect of DHEA compared with placebo at 3 months. Findings to date suggest that DHEA replacement seems to be well tolerated with an absence of significant side-effects. REVIEWER'S CONCLUSIONS: The data offer no support at present for an improvement in memory or other aspects of cognitive function following DHEA treatment in normal older people. In view of the growing public enthusiasm for DHEA supplementation, particularly in the USA, and the possibility that any neuroprotective effect of DHEA/S may only be evident in the long term, there is a need to undertake high quality trials in which the duration of DHEA treatment is longer than one year, and the number of participants is large enough to detect effects if they exist. Recently, trials of DHEA supplementation in Alzheimer's Disease (USA), post-menopausal women (USA), normal older men (UK), and a one-year trial in normal older men and women (France) have been completed. As soon as the results are available these studies will be included in the review. PMID- 11405959 TI - Conservative interventions for treating distal radial fractures in adults. AB - BACKGROUND: Fracture of the distal radius is a common clinical problem particularly in elderly white women with osteoporosis. OBJECTIVES: To determine the most appropriate conservative treatment for fractures of the distal radius in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group trials register, MEDLINE, PubMed, CINAHL, CENTRAL (Issue 2, 2000 of The Cochrane Library), the National Research Register (Issues 2 to 4, 2000) and bibliographies of trial reports. The search for the update was concluded in July 2000. SELECTION CRITERIA: Randomised or quasi-randomised clinical trials involving skeletally mature patients with a fracture of the distal radius, which compared commonly applied conservative interventions for fracture fixation. These included the application of an external support (plaster cast or brace) and fracture manipulation. DATA COLLECTION AND ANALYSIS: All trials, judged as fitting the selection criteria by both reviewers, were independently assessed by both reviewers for methodological quality. Data were extracted for anatomical, functional and clinical, including complications, outcomes. The trials were grouped into categories relating to cast position, extent and duration of immobilisation, use of braces, cast material and fracture manipulation. Although quantitative data from some trials are presented, the lack of good quality trials and trial heterogeneity inhibited pooling of results. MAIN RESULTS: In this update, three more trials were included and one previously included trial, excluded. In all, there were 31 trials, involving a total of 3372 mainly female and older patients, meeting the inclusion criteria for this review. Comprehensive details of the individual trials are provided in tabular form, and their results, grouped as indicated above, have been presented in text and analyses tables. The poor quality and heterogeneity in terms of patient characteristics, interventions compared and outcome measurement, of the included trials meant that no meta analyses were undertaken. REVIEWER'S CONCLUSIONS: There remains insufficient evidence from randomised trials to determine which methods of conservative treatment are the most appropriate for the more common types of distal radial fractures in adults. Therefore, at present, practitioners applying conservative management should use an accepted technique with which they are familiar, and which is cost-effective from the perspective of their provider unit. Patient preferences and circumstances, and the risk of complications should also be considered. Further research to clarify the most appropriate conservative treatment for this common fracture is warranted but requires careful preparation with an emphasis on undertaking a systematic approach. For this, it is important to reflect on the issues raised in this review, to ascertain patient preferences, prioritise researchable questions and agree a core data set for classification of fractures and outcomes. PMID- 11405960 TI - Extramedullary fixation implants for extracapsular hip fractures. AB - BACKGROUND: Extramedullary fixation of hip fractures refers to the application of a plate and screws to the lateral side of the proximal femur. OBJECTIVES: To compare different types of extramedullary fixation implants for the surgical treatment of extracapsular hip fracture in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register and reference lists of relevant articles. Date of the most recent search: January 2001. SELECTION CRITERIA: All randomised or quasi-randomised trials comparing extramedullary implants used in the fixation of extracapsular hip fracture in adults. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, using a ten item scale, and extracted data. Additional information was sought from all trialists. Wherever appropriate and possible, data from comparable trials were pooled. MAIN RESULTS: A newly identified trial investigating the RAB plate (a fixed angle blade plate with an oblique connecting strut) is included in this update. All eight included trials (involving 1173 participants) had methodological flaws which may have resulted in serious bias. Three trials involving 355 patients compared a fixed nail plate (Jewett or McLaughlin) with the sliding hip screw (SHS). The limited data presented indicated an increased risk of fixation failure outcomes for fixed nail plates. The two trials, involving 433 patients, comparing the RAB plate with the SHS had contrasting results, notably in terms of operative complications, fixation failure and anatomical restoration. One trial involving 100 patients compared the Pugh nail and the SHS. There was no significant difference between implants for the outcome measures reported. One trial involving 178 patients with 182 fractures, compared the Medoff plate with the SHS. A significantly higher mean operative blood loss and longer mean operative time were reported for the Medoff plate. There was however a tendency to a lower risk of fixation failure for unstable trochanteric fractures fixed with the Medoff plate. One trial involving 107 patients with subtrochanteric fractures compared the Medoff plate with three different screw plate systems. There was a tendency to a lower fixation failure rate for the Medoff plate, but no evidence for differences in longer-term outcomes. REVIEWER'S CONCLUSIONS: The fixed nail plate has higher risks of implant breakage and fixation failure than the SHS. Though insufficient evidence on other outcomes is available from randomised trials, the increased fixation failure rate is a major consideration and thus the SHS appears preferable. Insufficient information is available to draw firm conclusions of the clinical significance of differences between the SHS and either the RAB plate, the Pugh nail or the Medoff plate. PMID- 11405961 TI - Hydergine for dementia. AB - BACKGROUND: Currently hydergine is used almost exclusively for treating patients with either dementia, or 'age-related' cognitive symptoms. Since the early eighties there have been over a dozen more clinical trials, yet hydergine's efficacy remains uncertain. Although previous reviews offer generally favorable support for hydergine's efficacy, they were, however, limited by a bias with respect to the particular clinical studies chosen (eg, the inclusion of case reports, and uncontrolled trials), and by authors' impressionistic assessments of results. Not surprisingly, there has been a lack of consensus among reviewers with regard to the efficacy of hydergine. In 1994, a meta-analysis was published by the present reviewers who reported that overall, hydergine was more effective than placebo. However they also observed that the statistical evidence for efficacy in 'possible or probable Alzheimer's disease' patients was so modest that one additional statistically non-significant trial would have reduced the results to non significance. OBJECTIVES: Because of uncertainty surrounding the efficacy of hydergine, the goals of this overview were to assess its overall effect in patients with possible dementia, and to investigate potential moderators of an effect. SEARCH STRATEGY: The trials were identified from a search of the Specialised Register of the Cochrane Dementia and Cognitive Improvement Group on 15 November 2000 using the terms hydergin*, ergoloid* and dihydroergo*. Two proprietary databases were searched also. Published reviews were inspected for further sources. SELECTION CRITERIA: Trials to be included must be randomized, double-blind, parallel-group, and unconfounded comparisons of hydergine with placebo for a treatment duration of greater than 1 week in subjects with dementia or symptoms consistent with dementia. DATA COLLECTION AND ANALYSIS: Data were extracted independently by the reviewers, pooled where appropriate and possible, and the pooled odds ratios (95%CI) or the average differences (95%CI) were estimated. Where possible, intention-to-treat data were used. Outcomes of interest included clinical global impressions of change and comprehensive rating scales. Potential moderating variables of a treatment effect included: inpatient/outpatient status, trial duration, age, sex, medication dose, publication year, and diagnostic grouping. MAIN RESULTS: There were a total of nineteen trials that met inclusion criteria and that had data sufficient for analysis. Thirteen trials reported sufficient information to use a global rating of improvement and nine trials provided information on a comprehensive rating scale. Three trials provided both outcome measures. It was not possible to use many of the published results in a combined analysis owing to the lack of sufficient data to perform statistical analyses. For the twelve trials that used global ratings, there was a significant effect favoring hydergine (OR 3.78, 95%CI, 2.72-5.27). For the nine trials that used comprehensive ratings, there was a significant mean difference favoring hydergine (WMD 0.96, 95%CI, 0.54-1.37). Hydergine was well tolerated in these trials, with 78% of randomized subjects available for data analyses. Greater effect sizes on global ratings were associated with younger age, and possibly higher dose, although most of the subgroup analyses were statistically insignificant. REVIEWER'S CONCLUSIONS: As in an earlier systematic review, we found hydergine to show significant treatment effects when assessed by either global ratings or comprehensive rating scales (based here on a smaller set of trials than in the earlier published systematic review because trials were required to have data that could conform with MetaView, the Cochrane Collaboration statistics software). The small number of trials available for analysis, however, limited the ability of subgroup analyses to identify statistically significant moderating effects. Unfortunately, most of the randomized, double-blind, and placebo-controlled trials of hydergine were conducted and published before the advent of consensus-based diagnostic standards of dementia in 1984; therefore diagnostic criteria were less specific. As a result, uncertainty remains regarding hydergine's efficacy in dementia. PMID- 11405962 TI - Intravenous immunoglobulin for preventing infection in preterm and/or low-birth weight infants. AB - BACKGROUND: Nosocomial infections continue to be a significant cause of morbidity and mortality among preterm and/or low birth weight infants. Maternal transport of immunoglobulins to the fetus mainly occurs after 32 weeks gestation and endogenous synthesis does not begin until several months after birth. Administration of intravenous immunoglobulin provides IgG that can bind to cell surface receptors, provide opsonic activity, activate complement, promote antibody dependent cytotoxicity, and improve neutrophilic chemoluminescence. Intravenous immunoglobulin thus has the potential of preventing or altering the course of nosocomial infections. OBJECTIVES: To assess the effectiveness/safety of intravenous immunoglobulin (IVIG) administration (compared to placebo or no intervention) to preterm (< 37 weeks gestational age at birth) and/or low birth weight (LBW) (< 2500 g BW) infants in preventing nosocomial infections. SEARCH STRATEGY: MEDLINE, EMBASE, and the Cochrane Library Databases were searched in February 2001 using the keywords: immunoglobulin and infant-newborn and random allocation or controlled trial or randomized controlled trial (RCT). The reference lists of identified RCTs and personal files were searched. No language restrictions were applied. SELECTION CRITERIA: The criteria used to select studies for inclusion in this overview were: 1) DESIGN: RCTs in which administration of IVIG was compared to a control group that received a placebo or no intervention. 2) POPULATION: preterm (< 37 weeks gestational age) and/or LBW (<2500 g) infants. 3) INTERVENTION: IVIG for the prevention of bacterial/fungal infection during initial hospital stay (8 days or longer). (Studies that were primarily designed to assess the effect of IVIG on humoral immune markers were excluded as were studies in which the follow-up period was one week or less). 4) At least one of the following outcomes was reported: sepsis, any serious infection, death from all causes, death from infection, length of hospital stay, intraventricular haemorrhage (IVH), necrotizing enterocolitis (NEC), bronchopulmonary dysplasia (BPD). DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted information for each outcome reported in each study, and one researcher (AO) checked for any discrepancies and pooled the results. Relative risk (RR) and Risk Difference (RD) with 95% confidence intervals (CI) using the fixed effects model are reported. When a statistically significant RD was found the number needed to treat (NNT) was also calculated with 95% CIs. The results include all accepted studies in which the outcome of interest was reported. Statistically significant between study heterogeneity was reported. MAIN RESULTS: Nineteen studies met inclusion criteria. These included aprox. 5,000 preterm and/or LBW infants and reported on at least one of the outcomes of interest for this systematic review. When all studies were combined there was a statistically significant reduction (p = 0.02) in sepsis, RR [0.85 (95% CI 0.74, 0.98)] and RD [-0.03 (95% CI 0.00, -0.05)], NNT 33. There was statistically significant between-study heterogeneity (p = 0.02). A statistically significant reduction was found for any serious infection, one or more episodes, when all studies were combined [RR 0.82 (95% CI 0.74, 0.92); RD -0.04 (95% CI -0.02, 0.06,); NNT 25 (95% CI, 16.7, 50). There was statistically significant between study heterogeneity (p = 0.0006). There were no statistically significant differences for mortality from all causes, mortality from infection, incidence of NEC, BPD and IVH or length of hospital stay. No major adverse effects of IVIG were reported in any of the studies. REVIEWER'S CONCLUSIONS: IVIG administration results in a 3% reduction in sepsis and a 4 % reduction in any serious infection, one or more episodes, but is not associated with reductions in other important outcomes: sepsis, NEC, IVH, or length of hospital stay. Most importantly IVIG administration does not have any effect on mortality from any cause or from infections. Prophylactic use of IVIG is not associated with any short term serious side effects. From a clinical perspective a 3-4% reduction in nosocomial infections without a reduction in mortality or other important clinical outcomes is of marginal importance. The decision to use prophylactic IVIG will depend on the costs and the values assigned to the clinical outcomes. There is no justification for further RCTs testing the efficacy of previously studied IVIG preparations to reduce nosocomial infections in preterm and/or LBW infants. The results of these meta-analyses should encourage basic scientists and clinicians to pursue other avenues to prevent nosocomial infections. PMID- 11405963 TI - Nitric oxide for respiratory failure in infants born at or near term. AB - BACKGROUND: Nitric oxide is a major endogenous regulator of vascular tone. Inhaled nitric oxide gas has been investigated as a treatment for persistent pulmonary hypertension of the newborn. OBJECTIVES: To determine whether treatment of hypoxemic term and near-term newborn infants with inhaled nitric oxide (iNO) improves oxygenation and reduces the rates of death, the requirement for extracorporeal membrane oxygenation (ECMO), or affects long term neurodevelopmental outcomes. SEARCH STRATEGY: Electronic and hand searching of pediatric/neonatal literature and personal data files. In addition we contacted the principal investigators of articles which have been published as abstracts to ascertain the necessary information. SELECTION CRITERIA: Randomized and quasi randomized studies of inhaled nitric oxide in term and near term infants with hypoxic respiratory failure. Clinically relevant outcomes, including death, requirement for ECMO, and oxygenation. DATA COLLECTION AND ANALYSIS: Trial reports were analyzed for methodologic quality using the criteria of the Cochrane Neonatal Review Group. Results of mortality, oxygenation, short term clinical outcomes (particularly need for ECMO), and long term developmental outcomes were tabulated. STATISTICS: For categorical outcomes, typical estimates for relative risk and risk difference were calculated. For continuous variables, typical estimates for weighted mean difference were calculated. 95% confidence intervals were used. A fixed effect model was assumed for meta-analysis. MAIN RESULTS: Eleven eligible randomized controlled studies were found in term and near term infants with hypoxia. Entry criteria were reasonably consistent except for the one trial that studied only infants with congenital diaphragmatic hernia (Ninos 1997), and one trial that enrolled both preterm and term infants (Mercier 1998), but which reported the majority of the results separately for the two groups. Inhaled nitric oxide appears to improve outcome in hypoxemic term and near term infants by reducing the incidence of the combined endpoint of death or need for ECMO. The reduction seems to be entirely a reduction in need for ECMO; mortality is not reduced. Oxygenation improves in approximately 50% of infants receiving nitric oxide. The Oxygenation Index decreases by a (weighted) mean of 15.1 within 30 to 60 minutes after commencing therapy and PaO2 increases by a mean of 53 mmHg. Whether infants have clear echocardiographic evidence of PPHN or not does not appear to affect outcome. The outcome of infants with diaphragmatic hernia was not improved; indeed there is a suggestion that outcome was slightly worsened. The incidence of disability, incidence of deafness and infant development scores are all similar between tested survivors who received nitric oxide or not. REVIEWER'S CONCLUSIONS: On the evidence presently available, it appears reasonable to use inhaled nitric oxide in an initial concentration of 20 ppm for term and near term infants with hypoxic respiratory failure who do not have a diaphragmatic hernia. PMID- 11405964 TI - Membrane sweeping for induction of labour. AB - BACKGROUND: This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. Sweeping of the membranes, also commonly named stripping of the membranes, is a relatively simple technique usually performed without admission to hospital. During vaginal examination, the clinician's finger is introduced into the cervical os. Then, the inferior pole of the membranes is detached from the lower uterine segment by a circular movement of the examining finger. This intervention has the potential to initiate labour by increasing local production of prostaglandins and, thus, reduce pregnancy duration or pre-empt formal induction of labour with either oxytocin, prostaglandins or amniotomy. OBJECTIVES: To determine the effects of membrane sweeping for third trimester induction of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies of relevant papers (last searched November 2000). SELECTION CRITERIA: The criteria for inclusion included the following: (1) clinical trials comparing membrane sweeping used for third trimester labour induction with no vaginal examination or vaginal examination for cervical assessment only or with other methods listed above it on a predefined list of labour induction methods (i.e. administration of prostaglandins and oxytocin); (2) random allocation to the treatment or control group; (3) adequate or unclear allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: The data extraction was done centrally and incorporated into a series of reviews arranged by methods of induction of labour, following a standardised methodology. Two of the reviewers also assessed trial quality and extracted data. To avoid duplication of data in the reviews, the labour induction methods have been listed in a specific order, from one to 25. Each review includes comparisons between one of the methods with only those methods above it on the list. Therefore, sweeping of membranes was compared to no treatment, intravaginal prostaglandins and oxytocin. Results are reported as relative risk (RR) and their 95% confidence interval (CI) and number-needed-to-treat (NNT). MAIN RESULTS: Nineteen trials were included, 17 comparing sweeping of membranes with no treatment, three comparing sweeping with prostaglandins and one comparing sweeping with oxytocin (two studies reported more than one comparison). Risk of caesarean section was similar between groups (RR 0.97, 95% CI 0.73 to 1.28). Sweeping of the membranes, performed as a general policy in women at term, was associated with reduced duration of pregnancy and reduced frequency of pregnancy continuing beyond 41 weeks (RR 0.62, 95% CI 0.49 to 0.79) and 42 weeks (RR 0.28, 95% CI 0.15 to 0.50). To avoid one formal induction of labour, sweeping of membranes must be performed in seven women (NNT = 7). There was no evidence of a difference in the risk of maternal or neonatal infection. Discomfort during vaginal examination and other adverse effects (bleeding, irregular contractions) were more frequently reported by women allocated to sweeping. Studies comparing sweeping with prostaglandin administration are of limited sample size and do not provide evidence of benefit. REVIEWER'S CONCLUSIONS: Routine use of sweeping of membranes from 38 weeks of pregnancy onwards does not seem to produce clinically important benefits. When used as a means for induction of labour, the reduction in the use of more formal methods of induction needs to be balanced against women's discomfort and other adverse effects. PMID- 11405965 TI - Antibiotics for asymptomatic bacteriuria in pregnancy. AB - BACKGROUND: Up to 30% of mothers develop acute pyelonephritis if asymptomatic bacteriuria is untreated. Asymptomatic bacteriuria may have a role in preterm birth or it may be a marker for low socioeconomic status which is associated with low birth weight. OBJECTIVES: The objective of this review was to assess the effect of antibiotic treatment for asymptomatic bacteriuria on persistent bacteriuria during pregnancy, the risk of preterm delivery, and the development of pyelonephritis. SEARCH STRATEGY: I searched the Cochrane Pregnancy and Childbirth Group trials register. Date of last search: December 2000. SELECTION CRITERIA: Randomised trials comparing antibiotic treatment with placebo or no treatment in pregnant women with asymptomatic bacteriuria found on antenatal screening. DATA COLLECTION AND ANALYSIS: Trial quality was assessed. MAIN RESULTS: Fourteen studies were included. Overall the study quality was not strong. Antibiotic treatment compared to placebo or no treatment was effective in clearing asymptomatic bacteriuria (odds ratio 0.07, 95% confidence interval 0.05 to 0.10). The incidence of pyelonephritis was reduced (odds ratio 0.24, 95% confidence interval 0.19 to 0.32). Antibiotic treatment was also associated with a reduction in the incidence of preterm delivery or low birth weight babies (odds ratio 0.60, 95% confidence interval 0.45 to 0.80). REVIEWER'S CONCLUSIONS: Antibiotic treatment is effective in reducing the risk of pyelonephritis in pregnancy. An apparent reduction in preterm delivery is consistent with current theories about the role of infection in preterm birth, but this association should be interpreted with caution. PMID- 11405966 TI - Prophylactic versus selective use of surfactant in preventing morbidity and mortality in preterm infants. AB - BACKGROUND: Clinical trials have proven that surfactant therapy is effective in improving the immediate need for respiratory support and the clinical outcome of premature newborns (Soll 1992, Jobe 1993). Trials have studied a wide variety of surfactant preparations used either to prevent (prophylactic or delivery room administration) or treat (selective or rescue administration) respiratory distress syndrome. Using either treatment strategy, significant reductions in the incidence of pneumothorax, as well as significant improvement in survival, have been noted. It is unclear if there is an advantage to choosing either the prophylactic or selective approach to treatment OBJECTIVES: To compare the effect of prophylactic surfactant administration to surfactant treatment of established respiratory distress syndrome in premature infants. SEARCH STRATEGY: Searches were made of the Oxford Database of Perinatal Trials, Medline (MeSH terms: pulmonary surfactant; limits: age groups, newborn infants; publication type, clinical trials), previous reviews including cross-references, abstracts, conference and symposia proceedings, expert informants, and journal handsearching in the English language. SELECTION CRITERIA: Randomized controlled trials that compared the effects of prophylactic surfactant administration to surfactant treatment of established respiratory distress syndrome in premature infants were included in the analysis. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including the incidence of pneumothorax, pulmonary interstitial emphysema, patent ductus arteriosus, necrotizing enterocolitis, intraventricular hemorrhage (any grade and severe intraventricular hemorrhage), bronchopulmonary dysplasia, mortality, bronchopulmonary dysplasia or death, and retinopathy of prematurity were excerpted from the reports of the clinical trials by the reviewers. Data analysis was done in accordance with the standards of the Cochrane Neonatal Review Group. MAIN RESULTS: Eight studies were identified that met inclusion criteria. The majority of included studies noted an initial improvement in the respiratory status and a decrease in the incidence of respiratory distress syndrome in infants who received prophylactic surfactant. The meta-analysis supports a decrease in the incidence of pneumothorax, a decrease in the incidence of pulmonary interstitial emphysema, a decrease in the incidence of mortality and a decrease in the incidence of bronchopulmonary dysplasia or death associated with prophylactic administration of surfactant. No significant untoward effects of prophylactic surfactant administration are noted. In a secondary analysis of infants less than 30 weeks gestation, the meta analysis suggests a significant decrease in the risk of neonatal mortality and the risk of mortality or bronchopulmonary dysplasia. REVIEWER'S CONCLUSIONS: Prophylactic surfactant administration to infants judged to be at risk of developing respiratory distress syndrome (infants less than 30-32 weeks gestation), compared to selective use of surfactant in infants with established RDS, has been demonstrated to improve clinical outcome. Infants who receive prophylactic surfactant have a decreased risk of pneumothorax, a decreased risk of pulmonary interstitial emphysema and a decreased risk of mortality. However, it remains unclear exactly which criteria should be used to judge "at risk" infants who would require prophylactic surfactant administration. PMID- 11405967 TI - Interventions for isolated diaphyseal fractures of the ulna in adults. AB - BACKGROUND: Isolated fractures of the shaft of the ulna, which are often sustained when the forearm is raised to shield against a blow, are generally treated on an out-patient basis. OBJECTIVES: To assess the effects of various forms of treatment for isolated fractures of the ulnar shaft in adults. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group's specialised register (December 2000), the Cochrane Controlled Trials Register (The Cochrane Library Issue 4, 2000), MEDLINE (1966 to December 2000), EMBASE (1981 to December 2000), CINAHL (1982 to December 2000), and bibliographies of trial reports. Date of the most recent search: December 2000. SELECTION CRITERIA: Randomised or quasi randomised trials of conservative and surgical treatment of isolated fractures of the ulnar shaft in adults. Excluded were fractures of the proximal ulna and Monteggia fracture dislocations. DATA COLLECTION AND ANALYSIS: Independent quality assessment and data extraction were performed by all reviewers. Requests for more information were sent to trialists. Given the limited and poor quality evidence available, quantitative analysis was kept to a minimum. MAIN RESULTS: In the second update of this review, the only newly identified trial was excluded. Thus, as before, two small trials of conservative treatment, involving a total of 106 patients were included in this review. Both trials were of poor quality. One randomised trial compared short arm pre-fabricated functional braces with long arm plaster casts. There was no statistically significant difference in the time it took for fracture union. Patient satisfaction and return to work were better in the brace group. The other quasi-randomised trial compared Ace Wrap elastic bandage, short arm plaster cast and long arm plaster cast. The large loss to follow-up in this trial makes any data analysis tentative. However the need for replacement of the Ace wrap by other methods due to pain does indicate the potential for a serious problem with this intervention. REVIEWER'S CONCLUSIONS: There is insufficient evidence from randomised trials to determine which method of treatment is the most appropriate for isolated fractures of the ulnar shaft in adults. There is a need to establish the incidence of this injury and the outcome and associated costs of the various forms of treatment. Well designed randomised trials of current forms of conservative treatment are recommended. PMID- 11405968 TI - Pre-operative GnRH analogue therapy before hysterectomy or myomectomy for uterine fibroids. AB - BACKGROUND: Uterine fibroids, smooth muscle tumours of the uterus, are found in at least 25 to 35% of women over the age of 35 years. Although some of these tumours are asymptomatic, up to 50% cause symptoms severe enough to warrant therapy and surgery is the standard treatment. Fibroid growth is stimulated by oestrogen and gonadotropin releasing hormone agonists (GnRHa) which induce a state of hypoestrogenism have been investigated as a potential treatment. GnRHa treatment causes fibroids to shrink but cannot be used long term because of unacceptable symptoms and bone loss. Therefore, GnRHa may be useful pre operatively both to reduce fibroid and uterine volume and control bleeding. OBJECTIVES: The objective of this review is to evaluate the role of pre-treatment with gonadotropin releasing hormone (GnRH) analogues prior to a major surgical procedure, either hysterectomy or myomectomy, for uterine fibroids. SEARCH STRATEGY: Electronic searches for relevant randomised controlled trials of the Cochrane Menstrual Disorders and Subfertility Group Register of Trials, MEDLINE, EMBASE, the National Research Register, the National Library of Medicine's Clinical Trials Register and Current Contents were performed. Attempts were also made to identify published trials from citation lists of review articles and direct contact with drug companies for unpublished trials. In most cases, the first author of each included trial was contacted for additional information. The search was updated in October 2000. SELECTION CRITERIA: The inclusion criteria were randomised comparisons of GnRH analogue treatment versus placebo, no treatment, or other medical therapy prior to surgery, either myomectomy or hysterectomy, for uterine fibroids. DATA COLLECTION AND ANALYSIS: Twenty-six RCTs were identified that fulfilled the inclusion criteria for this review. The reviewers extracted the data independently and odds ratios for dichotomous outcomes and weighted mean differences for continuous outcomes were estimated from the data of fourteen trials where GnRH analogue treatment was compared with no pre-treatment and six trials where GnRH analogue treatment was compared with placebo. Three trials are awaiting assessment because the data were not in a suitable form for extraction or they are awaiting translation. Two trials have been excluded because the data were not in a suitable form for extraction and the authors were not able to provide additional information. One RCT compared GnRHa pre-treatment with lynestrenol pre-treatment. Results from pre-operative outcomes were combined for both types of surgery but results from intra- and post operative outcomes were reported separately for myomectomy and hysterectomy. Subgroup analysis was performed according to type of control group, no pre treatment or placebo, and for some outcomes there were additional subgroup analyses according to size of the uterus in gestational weeks. MAIN RESULTS: Pre- and post-operative haemoglobin (Hb) and haematocrit (HCT) were significantly improved by GnRH analogue therapy prior to surgery, and uterine volume, uterine gestational size and fibroid volume were all reduced. Pelvic symptoms were also reduced but some adverse events were more likely during GnRH analogue therapy. Hysterectomy appeared to be easier after pre-treatment with GnRH analogue therapy; there was reduced operating time and a greater proportion of hysterectomy patients were able to have a vaginal rather than an abdominal procedure. Duration of hospital stay was also reduced. Blood loss and rate of vertical incisions were reduced for both myomectomy and hysterectomy. Evidence of increased risk of fibroid recurrence after GnRH analogue pre-treatment in myomectomy patients was equivocal and few data were available to assess change in post-operative fertility. Lynestrenol did not offer any advantage over GnRH analogue therapy before fibroid surgery. The increased costs associated with GnRH analogue therapy were not assessed. REVIEWER'S CONCLUSIONS: The use of GnRH analogues for 3 to 4 months prior to fibroid surgery reduce both uterine volume and fibroid size. They are beneficial in the correction of pre-operative iron deficiency anaemia, if present, and reduce intra-operative blood loss. If uterine size is such that a mid-line incision is planned, this can be avoided in many women with the use of GnRH analogues. For patients undergoing hysterectomy, a vaginal procedure is more likely following the use of these agents. PMID- 11405969 TI - Antidepressant versus placebo for depressed elderly. AB - BACKGROUND: Depression warranting intervention is found in ten percent of people over the age of 60. Older depressed people are more likely to die than non depressed. Relatively few receive therapeutic interventions, and those that do, tend to receive low dose antidepressant therapy. Depression in older people is thought to differ in terms of aetiology, presentation, treatment and outcome than in younger people. Concomitant physical illness and increasing social, physical and neurophysiological diversity are associated with the ageing process. Consequently drug treatment of older patients is often carried out in institutions and on patients suffering from multiple physical problems. OBJECTIVES: To determine the efficacy of antidepressant medication compared with placebo in the treatment of depression in older patients. SEARCH STRATEGY: The search strategy incorporated: electronic literature searches of databases held by the Cochrane Collaboration Depression, Anxiety and Neurosis Review Group (CCDAN) (see Collaborative Review Group Search Strategy). Reference lists of related reviews and references of located studies. Contact was made with authors working in the field. SELECTION CRITERIA: All randomised, placebo controlled trials using antidepressants in the treatment of the presenting episode of depression in patients described as elderly, geriatric senile or older adult. DATA COLLECTION AND ANALYSIS: Two types of data were extracted (if available) from each study. The first type of data was dichotomous data, this consisted of recovered/not recovered. The second, continuous data,included: Hamilton Depression Rating Scale (HAM-D), Montgomery-Asberg Rating Scale (MADRS) and other depression rating scale scores. An analysis using Peto Odds ratios for the dichotomous data and weighted mean difference for continuous data was performed using RevMan 3.1. The presence of heterogeneity of treatment effect was assessed. MAIN RESULTS: Seventeen trials contributed data to the analyses comparing the efficacy of antidepressant treatment and placebo. Analyses of efficacy was based on 245 patients treated with TCAs (223 with placebo), 365 patients treated with SSRIs (372 with placebo) and 58 patients treated with MAOIs (63 with placebo). The standardised effect size for the three groups respectively were: TCAs; OR: 0.32 (0.21,0.47), SSRIs; OR; 0.51 (0.36,0.72), MAOIs: 0.17 (0.07,0.39). REVIEWER'S CONCLUSIONS: TCAs, SSRIs and MAOIs are effective in the treatment of older community patients and inpatients likely to have severe physical illness. At least six weeks of antidepressant treatment is recommended to achieve optimal therapeutic effect. There is little evidence concerning the efficacy of low dose TCA treatment. Further trials are required before low dose TCA treatment is routinely recommended. PMID- 11405970 TI - Improving health professionals' management and the organisation of care for overweight and obese people. AB - BACKGROUND: Obesity is increasing throughout the industrialised world. If left unchecked it will have major implications for both population health and costs to health services. OBJECTIVES: To assess whether health professionals' management or the organisation of care for overweight and obese people could be improved. SEARCH STRATEGY: We searched the specialised registers of the Cochrane Effective Practice and Organisation of Care Group (April 2000), the Cochrane Depression, Anxiety and Neurosis Group (August 1997), the Cochrane Diabetes Group (August 1997), the Cochrane Controlled Trials Register (September 1997), MEDLINE to April 2000, EMBASE to February 2000, Cinahl (1982 to February 2000), PsycLit (1974 to May 2000), Sigle (1980 to April 2000), Sociofile (1974 to October 1997), Dissertation Abstracts (1861 to January 1998), Conference Papers Index (1973 to January 1998), Resource Database in Continuing Medical Education. We also hand searched seven key journals and contacted experts in the field. SELECTION CRITERIA: Randomised trials, controlled before-and-after studies and interrupted time series analyses of providers' management of obesity or the organisation of care to improve provider practice or patient outcomes. We addressed three a priori comparisons and a fourth post hoc comparison. 1. Interventions aimed at improving health professionals' management or the delivery of health care for overweight/obese patients are more effective than usual care. 2. Interventions aimed at redressing negative attitudes and related practices towards overweight/obese patients are more effective than usual care. 3. Organisational interventions designed to change the structure of services for overweight/obese people are more effective than educational or behavioural interventions for health professionals. 4. Comparisons of different organisational interventions. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Eighteen studies were included involving more than 447 providers and 4104 patients. Six studies were identified for comparison 1. Five were professional-oriented interventions (the use of reminders and training) and the sixth was a study of professional and organisational interventions of shared care. No studies were identified for comparisons 2 or 3. Twelve studies were identified for post hoc comparison 4. These compared either the deliverer of weight loss interventions or the setting of interventions. The included studies were heterogeneous and of generally poor quality. REVIEWER'S CONCLUSIONS: At present, decisions about improving provision of services must be based on the evidence of patient interventions and good clinical judgement. Further research is needed to identify cost effective strategies for improving the management of obesity. PMID- 11405971 TI - Piracetam for dementia or cognitive impairment. AB - OBJECTIVES: To determine the clinical efficacy of piracetam for the features of dementia or cognitive impairment, classified according to the major subtypes of dementia: vascular, Alzheimer's disease or mixed vascular and Alzheimer's disease, or unclassified dementia, or cognitive impairment not fulfilling the criteria for dementia. SEARCH STRATEGY: The trials were identified from a search of the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group on 10 November 2000 using the term spiracetam, nootropic and 2-Oxo-1 pyrrolidine. In addition the pharmaceutical company responsible for marketing most of the piracetam worldwide, UCB Pharma, provided a comprehensive list of abstracts, which included many unpublished studies. As many of these unpublished, placebo-controlled studies will be reviewed as possible. SELECTION CRITERIA: All unconfounded trials specified as randomized in which treatment with piracetam was administered for more than a day and compared with placebo in patients with dementia of Alzheimer type, vascular dementia,or mixed vascular and Alzheimer's disease, or unclassified dementia, or cognitive impairment not fulfilling the criteria for dementia. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. Each study was independently verified as fulfilling the inclusion criteria. Studies were rated for methodological quality by assessment of blinding and loss before analysis as described by Jadad et al. (1996). Studies were pooled if appropriate and possible, and the pooled odds ratios (95%CI) or the average differences (95%CI) were estimated. Where possible, intention-to-treat analyses were undertaken. Sensitivity analyses were performed to determine if successive elimination of those studies performing most poorly on these quality criteria changed the effect estimate. MAIN RESULTS: Unfortunately, many of the studies were of cross-over design and first-phase data were unavailable, or could not be extracted. Global Impression of Change was the only outcoeme for which there was a significant volume of evidence from the pooled data. There was evidence of heterogeneity in the results from the individual studies, chi-square test = 20.8 (df=5). Using a fixed effects model the odds ratio for improvement in the piracetam group compared with the placebo group was 3.55, [95% CI][2.45, 5.16]. If a random effects model was used the odds ratio was 3.47 [1.29, 9.30]. If one single-blind study was excluded, the fixed effects model yielded an odds ratio of 3.36 [2.29, 4.99] and if a random effects model was applied then the odds ratio was 2.89 [1.01, 8.24]. The evidence of effects on cognition and other measures, was inconclusive. REVIEWER'S CONCLUSIONS: At this stage the evidence available from the published literature does not support the use of piracetam in the treatment of people with dementia or cognitive impairment. Although effects were found on global impression of change, no benefit was shown by any of the more specific measures. There is a need for further evaluation of piracetam by : 1) Obtaining the data from the identified studies for an individual patient database review, 2) Performing a randomized trial of piracetam in patients with diagnoses made by currently accepted diagnostic criteria. The trial should extend over for a period of at least 6 months and preferably longer. Specific cognitive instruments which are sensitive to change, Clinician Global Impression of Change, levels of dependency and caregiver quality of life scales should also be incorporated in such a study. PMID- 11405972 TI - Antidepressant plus benzodiazepine for major depression. AB - BACKGROUND: Anxiety frequently coexists with depression. Adding benzodiazepines to antidepressants is commonly used to treat people with depression, although there has been no convincing evidence to show that such a combination is more effective than antidepressants alone and that there are suggestions that benzodiazepines may lose their efficacy with long-term administration and that their chronic use carries risks of dependence. OBJECTIVES: To determine whether, among adult patients with major depression, adding benzodiazepines to antidepressants brings about any benefit in terms of symptomatic recovery or side effects in the short term (less than 8 weeks) and long term (more than 2 months), in comparison with treatment by antidepressants alone. SEARCH STRATEGY: We searched MEDLINE (1972 to September 1997), EMBASE (1980 to September 1997), International Pharmaceutical Abstracts (1972 to September 1997), Biological Abstracts (1984 to September 1997), LILACS (1980 to September 1997), PsycLIT (1974 to September 1997), the Cochrane Library (issue 3, 1997) and the trial register of the Cochrane Depression, Anxiety and Neurosis Group (last searched March 1999), combined with hand searching, reference searching, SciSearch and personal contacts. SELECTION CRITERIA: All randomised controlled trials that compared combined antidepressant-benzodiazepine treatment with antidepressant alone for adult patients with major depression. Exclusion criteria are: antidepressant dosage lower than 100 mg of imipramine or its equivalent daily and duration of trial shorter than four weeks. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the eligibility and quality of the studies. Two reviewers independently extracted the data. Standardized weighted mean differences and relative risks were estimated with random effects model. The dropouts were assigned the least favourable outcome. Two sensitivity analyses examined the effect of this assumption as well as the effect of including medium quality studies. Three a priori subgroup analyses were performed with regard to the patients with or without comorbid anxiety and with regard to the type. MAIN RESULTS: Aggregating nine studies with a total of 679 patients, the combination therapy group was less likely to drop out than the antidepressant alone group (relative risk 0.63, 95% confidence interval 0.49 to 0.81). The intention-to treat analysis (with people dropping out assigned the least favourable outcome) showed that the combination group was more likely to show improvement in their depression (defined as 50% or greater reduction in the depression scale from baseline) (relative risk 1.63, 95% confidence interval 1.18 to 2.27 at one week and relative risk 1.38, 95% confidence interval 1.15 to 1.66 at four weeks). The difference was no longer significant at six to eight weeks. None of the included RCTs lasted longer than eight weeks. The patients allocated to the combination therapy were less likely to drop out from the treatment due to side effects than those receiving antidepressants alone (relative risk 0.53, 95% confidence interval 0.32 to 0.86). However, these two groups of patients were equally likely to report at least one side effect (relative risk 0.99, 95% confidence interval 0.92 to 1.07). REVIEWER'S CONCLUSIONS: The potential benefits of adding a benzodiazepine to an antidepressant must be balanced judiciously against possible harms including development of dependence and accident proneness, on the one hand, and against continued suffering following no response and drop-out, on the other. PMID- 11405973 TI - Intravenous immunoglobulin for treating sepsis and septic shock. AB - OBJECTIVES: Death from severe sepsis and septic shock is common, and researchers have explored whether antibodies to the endotoxins in some bacteria reduces mortality. This review summarises the effects of intravenous immunoglobulin (IVIG) in patients with bacterial sepsis or septic shock on mortality, bacteriological failure rates, and duration of stay in hospital. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, MEDLINE 1966 to 2000, EMBASE 1988 to February 1999; we contacted investigators active in the field for unpublished data. SELECTION CRITERIA: Randomised trials comparing intravenous immunoglobulin (monoclonal or polyclonal) with placebo or no intervention, in patients with bacterial sepsis or septic shock. DATA COLLECTION AND ANALYSIS: Inclusion criteria, trial quality assessment, and data abstraction were done in duplicate. We conducted pre-specified subgroup analyses by type of immunoglobulin preparation. MAIN RESULTS: Twenty-seven out of 55 studies met our inclusion criteria. Pooled analysis of all types of IVIG preparations revealed a significant trend toward reduction of mortality (n= 8,856; RR=0.91; 95% CI 0.86 to 0.96). Overall mortality was reduced in patients who received polyclonal IVIG (n=492; RR=0.64; 95% CI 0.51 to 0.80). Mortality was not reduced among patients who received monoclonal antibodies such as anti-endotoxins (n=2,826 in 5 good quality studies; RR=0.97; 95% CI 0.88 to 1.07) or anti-cytokines (n=4,318 in 4 good quality studies; RR=0.93; 95% CI 0.86 to 1.01). A few studies measured secondary outcomes (deaths from sepsis or length of hospitalisation) but no differences in the intervention and control groups were identified except among those who received polyclonal IVIG, where sepsis-related mortality was significantly reduced (n=161; RR=0.35; 95% CI 0.18 to 0.69). REVIEWER'S CONCLUSIONS: In our opinion, polyclonal IVIG significantly reduces mortality and can be used as an adjuvant treatment for sepsis and septic shock. Adjunctive therapy with monoclonal IVIGs remains experimental. PMID- 11405974 TI - Interventions for treating constipation in pregnancy. AB - BACKGROUND: Constipation is a common problem in late pregnancy. Circulating progesterone may be the cause of slower gastrointestinal movement in mid and late pregnancy. OBJECTIVES: The objective of this review was to assess the effects of different methods for treating constipation in pregnancy. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and MEDLINE. Date of last search: January 2001. SELECTION CRITERIA: Randomised trials of any treatment for constipation in pregnancy. DATA COLLECTION AND ANALYSIS: Trial quality assessments and data extraction were done independently by two reviewers. MAIN RESULTS: Two suitable trials were identified. Fibre supplements increased the frequency of defecation (odds ratio 0.18, 95% confidence interval 0.05 to 0.67), and lead to softer stools. Stimulant laxatives are more effective than bulk-forming laxatives (odds ratio 0.30, 95% confidence interval 0.14 to 0.61), but may cause more side effects. REVIEWER'S CONCLUSIONS: Dietary supplements of fibre in the form of bran or wheat fibre are likely to help women experiencing constipation in pregnancy. If the problem fails to resolve, stimulant laxatives are likely to prove more effective. PMID- 11405975 TI - Delayed (>3 weeks) postnatal corticosteroids for chronic lung disease in preterm infants. AB - BACKGROUND: Many preterm babies who survive, having had respiratory distress syndrome (RDS) or not, go on to develop chronic lung disease (CLD). This is probably due to persistence of inflammation in the lung. Corticosteroids have powerful anti-inflammatory effects and have been used to treat established CLD. However it is unclear whether any beneficial effects outweigh the adverse effects of these drugs. OBJECTIVES: To determine if late (> 3 weeks) postnatal corticosteroid treatment vs control (placebo or nothing) is of benefit in the treatment of chronic lung disease (CLD) in the preterm infant. SEARCH STRATEGY: Randomised controlled trials of postnatal corticosteroid therapy were sought from the Oxford Database of Perinatal Trials, the Cochrane Controlled Trials Register, Medline, hand searching paediatric and perinatal journals, examining previous review articles and information received from practising neonatologists. SELECTION CRITERIA: Randomised controlled trials of postnatal corticosteroid treatment initiated at > 3 weeks of age in preterm infants with CLD were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding clinical outcomes including mortality before discharge, failure to extubate, infection, hyperglycaemia, hypertension, severe retinopathy of prematurity (ROP), necrotizing enterocolitis (NEC), gastrointestinal bleeding, intestinal perforation, echodensities on ultrasound scan of brain, need for home oxygen, glycosuria, need for late rescue with dexamethasone, cerebral palsy in survivors and blindness in survivors including a study reporting 5 year follow-up were abstracted and analysed using Revman 4.1. MAIN RESULTS: Delayed steroid treatment had no effect on mortality. Beneficial effects of delayed steroid treatment included reductions in failure to extubate by 7 or 28 days, need for late rescue treatment with dexamethasone, chronic lung disease at 36 weeks, death or CLD at 36 wk, and discharge to home on oxygen therapy. There was no increase in risk of infection, necrotising enterocolitis, or gastrointestinal bleeding. Short-term adverse affects included hyperglycaemia, glycosuria and hypertension. There was an increase in severe retinopathy of prematurity, of borderline significance in survivors, but no significant increase in blindness. Although there were increases in long-term neurologic sequelae, including abnormal neurologic examination and cerebral palsy, those apparent trends were offset by a trend in the opposite direction due to an increased incidence of death before follow-up assessment. REVIEWER'S CONCLUSIONS: The benefits of late corticosteroid therapy may not outweigh actual or potential adverse effects. Although there continues to be concern about an increased incidence of adverse neurological outcomes in infants treated with postnatal steroids (see also review of Early postnatal corticosteroids), this review of postnatal corticosteroid treatment for CLD initiated after three weeks of age suggests that late or delayed therapy may not significantly increase the risk of adverse long-term neurodevelopmental outcomes. Nonetheless, corticosteroids should still be reserved for infants who cannot be weaned from mechanical ventilation. The dose of dexamethasone and the duration of any course of treatment should still be minimized. PMID- 11405976 TI - Nerve blocks (subcostal, lateral cutaneous, femoral, triple, psoas) for hip fractures. AB - BACKGROUND: Various nerve blocks using local anaesthetic agents have been used in order to reduce pain after hip fracture. OBJECTIVES: To determine the effects of nerve blocks (inserted either pre-operatively, operatively or post-operatively) as part of the treatment for a hip fracture. SEARCH STRATEGY: The Cochrane Musculoskeletal Injuries Group specialised trials register, MEDLINE, and bibliographies of trial reports were searched. Date of the most recent search: October 2000. SELECTION CRITERIA: Randomised and quasi-randomised trials involving the use of nerve blocks as part of the care of a hip fracture patient. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, by use of a nine item scale, and extracted data. Wherever appropriate, results of outcome measures were pooled. MAIN RESULTS: Seven randomised or quasi-randomised trials involving 269 patients were included. Two trials related to insertion of a nerve block pre-operatively and the remaining five to peri-operative insertion. Nerve blocks resulted in a reduction of the quantity of parenteral or oral analgesia administered to control pain from the fracture/operation or during surgery and a reduction in reported pain levels. It was not possible to demonstrate if this reduction in analgesia use was associated with any other clinical benefit. REVIEWER'S CONCLUSIONS: Because of the small number of patients included in this review and the differing type of nerve blocks and timing of insertion, it is not possible to determine if nerve blocks confer any significant benefit when compared with other analgesic methods as part of the treatment of a hip fracture. Further trials with larger numbers of patients and full reporting of clinical outcomes would be justified. PMID- 11405977 TI - Interventions for treating head lice. AB - BACKGROUND: Infection with head lice is a widespread condition in developed and developing countries. Infection occurs most commonly in children, but also affects adults. If left untreated the condition can become intensely irritating and skin infections may occur if the bites are scratched. OBJECTIVES: The aim of this review was to assess the effects of interventions for head lice. SEARCH STRATEGY: Trials register of The Cochrane Infectious Diseases Group; Medline; Embase; Science Citation Index; Biosis and Toxline; reference lists of relevant articles; pharmaceutical companies producing pediculicides (published and unpublished trials); UK and US Regulatory Authorities. SELECTION CRITERIA: Randomised trials (published and unpublished) or trials using alternate allocation were sought which compared pediculicides with the same and different formulations of other pediculicides, and pediculicides with physical methods. DATA COLLECTION AND ANALYSIS: Of the 71 identified studies, only four met the inclusion criteria. Two reviewers independently assessed trial quality. One reviewer extracted the data. MAIN RESULTS: We found no evidence that any one pediculicide has greater effect than another. The two studies comparing malathion and permethrin with their respective vehicles showed a higher cure rate for the active ingredient than the vehicle. Another study comparing synergised pyrethrins with permethrin showed their effects to be equivalent. A comparative trial of malathion lotion vs combing, showed combing to be ineffective for the cureative treatment of head lice infection. Adverse effects were reported in a number of trials and were all minor, although reporting quality varied between trials. REVIEWER'S CONCLUSIONS: Permethrin, synergised pyrethrin and malathion were effective in the treatment of head lice. However, the emergence of drug resistance since these trials were conducted means there is no direct contemporary evidence of the comparative effectiveness of these products. The 'best' choice will now depend on local resistance patterns. Physical treatment methods(BugBusting) were shown ot be ineffective to treat head lice. No evidence exists regarding other chemical control methods such as the use of herbal treatments, when used in the curative treatment of head lice. Future trials should take into account the methodological recommendations that arise from this review. PMID- 11405978 TI - Amantadine and rimantadine for preventing and treating influenza A in adults. AB - BACKGROUND: Amantadine hydrochloride and rimantadine hydrochloride have antiviral properties, but they are not widely used due to a lack of knowledge of their properties and concerns about possible adverse effects. OBJECTIVES: The objective of this review was to assess the effects and safety of amantadine and rimantadine in healthy adults. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, Medline, Embase and reference lists of articles. We also contacted manufacturers, researchers and authors. SELECTION CRITERIA: Randomised and quasi randomised studies comparing amantadine and/or rimantadine with placebo, control antivirals or no intervention, or comparing doses or schedules of amantadine and/or rimantadine in healthy adults. DATA COLLECTION AND ANALYSIS: For prevention trials the numbers of participants with clinically defined influenza, with serologically confirmed clinical influenza A and adverse effects were analysed. Analysis for treatment trials was of the mean duration of fever and adverse effects. MAIN RESULTS: Amantadine prevented 23% of clinical influenza cases (95% confidence interval 11% to 34%), and 63% of serologically confirmed clinical influenza A cases (95% confidence interval 42% to 76%). Amantadine reduced duration of fever by one day (95% confidence interval 0.7 to 1.3). Rimantadine demonstrated comparable effectiveness, but there were fewer trials and the results for prevention were not statistically significant. Both amantadine and rimantadine induced significant gastrointestinal adverse effects. Adverse effects of the central nervous system adverse and study withdrawals were significantly more common with amantadine than rimantadine. REVIEWER'S CONCLUSIONS: Amantadine and rimantadine have comparable effectiveness in the prevention and treatment of influenza A in healthy adults, although rimantadine induces fewer adverse effects than amantadine. PMID- 11405979 TI - House dust mite control measures for asthma. AB - BACKGROUND: The major allergen in house dust comes from mites. Chemical, physical and combined methods of reducing mite allergen levels are intended to reduce asthma symptoms in people who are sensitive to house dust mites. OBJECTIVES: The objective of this review was to assess the effects of reducing exposure to house dust mite antigens in the homes of mite-sensitive asthmatics. SEARCH STRATEGY: We searched the Cochrane Airways Group trials register, checked reference lists of articles and hand-searched Respiration (1980 to 1996) and Clinical and Experimental Allergy (1980 to 1996). The Cochrane Library is searched every three months. SELECTION CRITERIA: Randomised trials of mite control measures vs placebo or no treatment in asthmatic people known to be sensitive to house dust mites. DATA COLLECTION AND ANALYSIS: Two reviewers applied the trial inclusion criteria, assessed their quality and extracted the data independently. Study authors were contacted to clarify information. MAIN RESULTS: Twenty-nine trials (939 patients in the analyses) were included, with two trials awaiting assessment. There was little difference in improvement of asthma between patients in experimental groups compared to control groups (relative risk 1.04, 95% confidence interval (95%CI) 0.83 to 1.31). Asthma symptom scores were also similar for the experimental and control groups (standardised mean difference (SMD) -0.07, 95% CI -0.35 to 0.22), however there was significant heterogeneity between studies p=0.015. This appears to have been due, in part, to the parallel group studies of physical treatments. These three studies (107 patients) showed a significant reduction in symptoms; SMD -0.44 (95% CI -0.83, -0.06) with no heterogeneity between the trials. No significant difference was noted for medication usage (SMD -0.14, 95%CI -0.43 to 0.15). Peak flow in the morning showed no significant difference between the experimental and the control groups (SMD 0.04, 95%CI -0.13 to 0.21). REVIEWER'S CONCLUSIONS: Current chemical methods aimed at reducing exposure to house dust mite allergens seem to be ineffective and cannot be recommended as prophylaxis for mite sensitive asthmatics. Physical reduction methods may reduce asthma symptoms, but results of larger and more rigorous studies are required before any recommendations can be made concerning this approach. PMID- 11405980 TI - Population-based interventions for reducing sexually transmitted infections, including HIV infection. AB - BACKGROUND: Sexually transmitted infections (STI) are common in developing countries. The World Health Organisation (WHO) estimates that in 1995, 333 million new cases of syphilis, gonorrhoea, chlamydial infection and trichomoniasis occurred. Human immunodeficiency virus (HIV) infection is also common in developing countries. UNAIDS estimates that over 90% of the 33 million people infected with HIV by December 1999 live in developing countries (UNAIDS 1999). The STI and HIV epidemics are interdependent. Similar behaviours, such as frequent unprotected intercourse with different partners, place people at high risk of both infections, and there is clear evidence that conventional STIs increase the likelihood of HIV transmission. Several studies have demonstrated a strong association between both ulcerative and non-ulcerative STIs, and HIV infection (Cameron 1989, Laga 1993) and there is biological evidence that the presence of an STI increases shedding of HIV and that STI treatment reduces HIV shedding (Cohen 1997, Robinson 1997). Therefore, STI control may have the potential to contribute substantially to HIV prevention. OBJECTIVES: To determine the impact of population-based STI interventions on the frequency of HIV infection, frequency of STIs and quality of STI management. SEARCH STRATEGY: The following electronic databases were searched for relevant randomised trials or reviews: 1) MEDLINE for the years 1966 to current using the search terms sexually transmitted diseases and human immunodeficiency virus infection 2) The Cochrane Database of Systematic Reviews, Database of Abstracts of Reviews of Effectiveness and the Cochrane Clinical Trials Register in the most recent issue of the Cochrane Library 3) The specialist register of trials maintained by the Cochrane Infectious Diseases Group. 4) EMBASE The abstracts of relevant conferences were searched and reference lists of all review articles and primary studies were scanned. Finally, authors of included trials and other experts in the field were contacted as appropriate. SELECTION CRITERIA: Randomised controlled trials in which the unit of randomisation is either a community or a treatment facility. Studies where individuals are randomised were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied the inclusion criteria to potential studies with any disagreements resolved by discussion. Trials were examined for completeness of reporting. The methodological quality of each trial was assessed by the same two reviewers with details recorded of randomisation method, blinding, use of intention-to-treat analysis and the number of patients lost to follow-up using standard guidelines of the Cochrane Infectious Diseases Group. MAIN RESULTS: Four trials were included. Frequency of HIV infection: In Rakai, after 3 rounds of treatment of all community members for STIs the rate ratio of incident HIV infection was 0.97 (95%CI 0.81 to 1.16), indicating no effect of the intervention. In Mwanza, the incidence of HIV infection in the intervention groups (strengthened syndromic management of STIs in primary care clinics) was 1.2% compared with 1.9% in the control groups (OR=0.58, 95% CI 0.42-0.70), corresponding to a 38% reduction (95%CI 15% to 55%) in HIV incidence in the intervention group. Frequency of STIs: In both Mwanza and Rakai, there was no significant reduction in gonorrhoea, chlamydia, urethritis, or reported STI symptoms among intervention communities. The prevalence ratio of syphilis between intervention and control groups in Rakai was 0.8 (95%CI 0.71-0.89), of trichmoniasis was 0.59 (0.38-0.91), and of bacterial vaginosis was 0.87 (0.74 1.02). In Mwanza, the prevalence of serologically diagnosed syphilis in the intervention community was 5% compared with 7% in the control community at the end of the trial (adjusted relative risk 0.71 (95%CI 0.54-0.93). Quality of treatment: In Lima, following training of pharmacy assistants in STI syndromic management, symptoms were recognised as being due to an STI in 65% of standardised simulated patients (SSPs) visiting intervention and 60% of SSPs visiting control pharmacies (p=0.35). Medication was offered without referral to a doctor in most cases (83% intervention and 78% control, p=0.61). Of those SSPs offered medication, only 1.4% that visited intervention pharmacies and only 0.7% of those that visited control pharmacies (p=0.57) were offered a recommended regimen. Similarly in only 15% and 16% of SSP visits respectively was any recommended drug offered. However, education and counseling were more likely to be given to SSPs visiting intervention pharmacies (40% vs 27%, p=0.01). No SSPs were given partner cards or condoms. In Hlabisa, following the intervention targeting primary care clinic nurses (strengthened STI syndromic management and provision of STI syndrome packets containing recommended drugs, condom, partner cards and patient information leaflets), SSPs were more likely to be given recommended drugs in intervention clinics (83% vs 12%, p <0.005) and more likely to be correctly case managed [given correct drugs, partner cards and condoms] (88% vs 50%, p <0.005). There were no significant difference in the proportions adequately counseled (68% vs 46%, p=0.06), experiencing good staff attitude (84% vs 58%, p=0.07), and being consulted in privacy (92% vs 86%, p=0.4). There was no strong evidence of any impact on treatment seeking behaviour, utilisation of services, or sexual behaviour in any of the four trials. REVIEWERS' CONCLUSIONS: There is limited evidence from randomised controlled trials for STI control as an effective HIV prevention strategy. Improved STI treatment services have been shown to reduce HIV incidence in an environment characterised by an emerging HIV epidemic (low and slowly rising prevalence), where STI treatment services are poor and where STIs are highly prevalent. There is no evidence for substantial benefit from treatment of all community members. There are however other compelling reasons why STI treatment services should be strengthened and the available evidence suggests that when an intervention is accepted it can substantially improve quality of services provided. Further community based randomised controlled trials that test a range of alternative STI control strategies are needed in a variety of different settings. Such trials should aim to measure a range of factors that include health seeking behaviour and quality of treatment as well as HIV, STI and other biological endpoints. PMID- 11405981 TI - Intravenous immunoglobulin for suspected or subsequently proven infection in neonates. AB - BACKGROUND: Congenital and nosocomial infections are important causes of neonatal morbidity and mortality. Maternal transport of immunoglobulins to the fetus mainly occurs after 32 weeks gestation and endogenous synthesis does not begin until several months after birth. Administration of intravenous immunoglobulin provides IgG that can bind to cell surface receptors, provide opsonic activity, activate complement, promote antibody dependent cytotoxicity, and improve neutrophilic chemo luminescence. Theoretically infectious morbidity and morbidity could be reduced by the administration of intravenous immunoglobulin. OBJECTIVES: To assess the effectiveness of intravenous immunoglobulin (IVIG) to reduce mortality/morbidity caused by suspected infection in newborn infants. In secondary analyses to assess the effectiveness of IVIG to reduce mortality/morbidity in those neonates who entered into the studies with suspected infection and who later were confirmed as being infected. SEARCH STRATEGY: MEDLINE, EMBASE and the Cochrane Library were searched in February 2001 using the following keywords: immunoglobulin and infant-newborn, and random allocation, or controlled trial, or randomized controlled trial (RCT). The reference lists of identified RCTs, meta-analyses and personal files were searched. No language restrictions were applied. Unpublished information was requested from and obtained from five researchers (~~ Erdem 1993~~; ~~ Gokalp 1994~~; ~~ Haque 1988~~; ~~ M-Ramirez 1992~~; ~~ Shenoi 1999~~). SELECTION CRITERIA: The criteria used to select studies for inclusion were: 1) DESIGN: RCT (including quasi randomized trials) 2) Newborn infants (< 28 days old) 3) INTERVENTION: IVIG for treatment of suspected (and in some infants subsequently proved) bacterial/fungal infection compared to placebo or no intervention. 4) At least one of the following outcomes was reported: mortality during initial hospital stay; length of hospital stay; side effects; psychomotor development/growth at follow up. DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted information for the outcomes of interest and one researcher (AO) checked for any discrepancies and pooled the results. Typical Relative Risk (RR) and Risk Difference (RD) with 95% confidence intervals (CI) using the fixed effects model are reported for dichotomous outcomes and weighted mean difference (WMD) for continuous data. NNT was calculated for outcomes that showed a statistically significant reduction in RD. MAIN RESULTS: Five hundred twenty nine neonates with suspected infection have been enrolled in RCTs to evaluate the effect of IVIG on neonatal outcomes. These studies were undertaken in seven countries. Six studies (n = 318) reported on the outcomes of randomized patients with clinically suspected infection. The results showed a reduction in mortality following IVIG treatment [typical RR 0.63 (95% CI; 0.40, 1.00), RD -0.09 (95% CI; 0.00, -0.17) of borderline statistical significance. Treatment with IVIG (seven trials, n = 262) in cases of subsequently proved infection did result in a statistically significant reduction in mortality [typical RR 0.55 (95% CI; 0.31, 0.98; RD -0.09 (95% CI; -0.01, 0.18); NNT 11 (95% CI; 5.6, 100]. In spite of different geographical locations of the studies, differences in the mortality in the control groups (range 0% - 43.8%), the use of different IVIG preparations, and different dosing regimens, there was no statistically significant between-study heterogeneity for the outcome of mortality in the two analyses. REVIEWER'S CONCLUSIONS: There is insufficient evidence to support the routine administration of IVIG preparations investigated to date to prevent mortality in infants with suspected or subsequently proved neonatal infection. Researchers should be encouraged to undertake well-designed trials to confirm or refute the effectiveness of IVIG to reduce adverse outcomes in neonates with suspected infection. PMID- 11405982 TI - Hip protectors for preventing hip fractures in the elderly. AB - BACKGROUND: Hip fracture in the elderly is usually the result of a simple fall. Hip protectors have been advocated as a means to reduce the risk of sustaining a fracture in a fall on the hip. OBJECTIVES: To determine if external hip protectors reduce the incidence of hip fractures in elderly persons following a fall. SEARCH STRATEGY: We searched the Cochrane Musculoskeletal Injuries Group specialised register, Cochrane Controlled Trials Register, MEDLINE, EMBASE, CINAHL and reference lists of relevant articles. Identified trialists were contacted. Date of the most recent search: December 2000. SELECTION CRITERIA: All randomised or quasi-randomised controlled trials comparing the use of hip protectors with a control group. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality, using a ten item scale, and extracted data. Additional information was sought from all trialists. Wherever appropriate and possible, the data are presented graphically. MAIN RESULTS: In this second update, a further trial has been included to bring the total number of included trials to seven (3553 participants). All studies involved elderly people in nursing homes or residential care or supported living at home, four within the Scandinavian countries, one in Japan, one in the United Kingdom and one in Australia. The three largest studies involving 3210 participants randomised by care unit, nursing home or nursing home ward rather than by the individual (cluster randomisation). One study of 141 individuals was primarily a compliance study lasting only 12 weeks. Summation of results from the other six studies gave an occurrence of hip fractures of 29/1313 (2.2%) for those allocated to wear hip protectors, against 130/2099 (6.2%) to those not allocated to wear protectors. However, due to the large number of participants allocated by cluster randomisation, it was not possible to determine if this difference between groups was statistically significant. No important adverse effects of the hip protectors were reported but compliance, particularly in the long term, was poor. REVIEWER'S CONCLUSIONS: Hip protectors appear to reduce the risk of hip fracture within a selected population at high risk of sustaining a hip fracture. The generalisation of the results is unknown beyond high-risk populations. Cost effectiveness is unclear. Results from ongoing trials may clarify this situation. Acceptability by users of the protectors remains a problem, due to discomfort and practicality. PMID- 11405983 TI - Interventions for treating melioidosis. AB - BACKGROUND: Melioidosis is an infectious disease caused by a bacterium (Burkholderia pseudomallei) found particularly in some areas in the tropics. It is a serious condition which can be fatal. Beta lactam antibiotics have dramatically reduced the risk of death, but mortality still remains high. OBJECTIVES: To summarise evidence from randomised trials on the effects of treatment regimens on death and relapse. SEARCH STRATEGY: Cochrane Controlled Trials Register, MEDLINE, EMBASE, BIOSIS from 1966 to September 2000 using MeSH terms "pseudomallei", "melioidosis" together with the terms "randomized controlled-trial", "random allocation"; reference lists in articles on melioidosis; contact with trialists. SELECTION CRITERIA: Randomised and quasi randomised trials assessing treatments in patients with melioidosis. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality was assessed by two reviewers independently. MAIN RESULTS: For intravenous therapy in the acute phase, we identified five trials with a total of 519 patients. Chloramphenicol, doxycycline, and co-trimoxazole (trimethoprim-sulphamethoxazole) combination regimens were associated with a mortality of 50% or more (two studies). Patients randomised to regimens that included ceftazidime were more likely to survive (relative risk [RR] 0.46, 95% confidence interval [CI] 0.30 to 0.71). When ceftazidime-containing regimens were compared with beta lactam or alternative beta lactamase inhibitor regimens such as co-amoxiclav (amoxycillin-clavulanic acid) and cefoperazone-sulbactam, mortality rates were similar (RR 1.10, 95% CI 0.83 to 1.46), as was the case in one trial of imipenem. For oral therapy in the maintenance phase, we found two trials of 188 participants. Results showed that treatment with the conventional regimen (chloramphenicol, doxycycline, and trimethoprim-sulphamethoxazole) resulted in fewer fatalities in patients compared to a regimen of amoxycillin-clavulanic acid and doxycycline alone. REVIEWER'S CONCLUSIONS: Regimens for the acute phase of illness should contain ceftazidime or imipenem. It is not yet clear if combinations of treatments in the early phase reduce relapse. For oral therapy after the acute phase of treatment, trials suggest that conventional four drug regimens can be used for treatment. PMID- 11405984 TI - Oral corticosteroids for acute exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: Systemic corticosteroids are widely used in the management of patients with acute exacerbations of COPD, in combination with other treatments. OBJECTIVES: To determine the effect of corticosteroids, administered either parenterally or orally, on the outcome in patients with acute exacerbations of COPD. SEARCH STRATEGY: An initial search was carried out using the Cochrane Airways Group COPD RCT register with additional studies sought in the bibliographies of randomised controlled trials and review articles. Authors of identified randomised controlled trials were contacted for other published and unpublished studies. SELECTION CRITERIA: Randomised controlled trials comparing corticosteroids, administered either parenterally or orally, with appropriate placebo. Other interventions were standardised e.g. bronchodilators, antibiotics. Clinical studies of acute asthma were excluded. DATA COLLECTION AND ANALYSIS: Data was extracted by one of the reviewers and sent to authors for verification. All trials were combined using Review Manager (version 4.1) for analysis. MAIN RESULTS: We have identified 7 studies that fulfilled the inclusion criteria. Outcomes were varied and few were common to all studies. The most commonly reported outcome, the FEV1 between 6 - 72 hours after treatment, showed a significant treatment benefit for corticosteroid over placebo treatment, weighted mean difference 120 ml 95% confidence intervals: 5, 190 ml. There were significantly fewer treatment failures in patients given corticosteroid treatment, but the number of studies reporting this outcome was smaller and there was significant heterogeneity between them. There was an increased likelihood of an adverse drug reaction with corticosteroid treatment. REVIEWER'S CONCLUSIONS: Treatment with oral or parenteral corticosteroids increases the rate of lung function improvement over the first 72 hours of an exacerbation of chronic obstructive pulmonary disease, but at a significantly increased risk of an adverse drug reaction. There is no evidence that this benefit is maintained after 72 hours, or that other outcomes are improved. PMID- 11405985 TI - Colloid solutions for fluid resuscitation. AB - BACKGROUND: Colloids are widely used in the replacement of fluid volume, however doubts remain as to their benefits. Different colloids vary in their molecular weight and therefore in the length of time they remain in the circulatory system. Because of this and their other characteristics, they may differ in their safety and efficacy. OBJECTIVES: To compare the effects of different colloid solutions in patients thought to need volume replacement. SEARCH STRATEGY: The Cochrane Injuries Group specialised register, The Cochrane Controlled trials register (all years), MEDLINE (1994-2000), EMBASE (1974-2000) were searched. Bibliographies of trials retrieved were searched, and drug companies manufacturing colloids were contacted for information. SELECTION CRITERIA: Randomised and quasi randomised trials comparing colloid solutions in critically ill and surgical patients thought to need volume replacement. The main outcomes measured were death, amount of whole blood transfused and incidence of adverse reactions. DATA COLLECTION AND ANALYSIS: Two authors independently extracted the data and assessed the quality of the trials. MAIN RESULTS: 52 trials met the inclusion criteria, with a total of 3311 participants. Quality of allocation concealment was judged to be adequate in 17 trials and poor or uncertain in 35. Deaths were obtained from 31 trials. For albumin or PPF versus hydroxyethyl starch (HES) 20 trials (n=1029) reported mortality. The pooled relative risk (RR) was 1.17 (95% CI 0.91, 1.50). For albumin or PPF versus gelatin four trials (n=542) reported mortality. The RR was 0.99 (0.69, 1.42). For gelatin vs HES six trials (n=597) reported mortality, RR was 0.96 (0.69, 1.33). RR was not estimable in the albumin vs dextran, gelatin vs dextran, and HES vs dextran groups. Thirty-one trials recorded the amount of blood transfused, however quantitative analysis was not possible due to skewness and variable reporting. Fifteen trials recorded adverse reactions, but none occurred. REVIEWER'S CONCLUSIONS: From this review, there is no evidence that one colloid solution is more effective or safe than any other, although the confidence intervals are wide and do not exclude clinically significant differences between colloids. Larger trials of fluid therapy are needed if clinically significant differences in mortality are to be detected or excluded. PMID- 11405986 TI - Antibiotic prophylaxis for intrauterine contraceptive device insertion. AB - BACKGROUND: Concern about the risk of upper genital tract infection (pelvic inflammatory disease) often limits use of the IUD, a highly effective contraceptive. Prophylactic antibiotic administration around the time of induced abortion significantly reduces the risk of postoperative endometritis.(Sawaya, 1996) Since the risk of IUD-related infection is limited to the first few weeks to months after insertion,(Lee, 1983; Farley, 1992) contamination of the endometrial cavity at the time of insertion(Mishell, 1966) appears to be the mechanism, rather than the IUD or string itself. Thus, antibiotic administration before IUD insertion might reduce the risk of upper genital tract infection from passive introduction of bacteria at insertion. OBJECTIVES: To assess the effectiveness of prophylactic antibiotic administration before IUD insertion in reducing IUD-related complications and discontinuations within three months of insertion. The primary outcome was pelvic inflammatory disease (four reports) or early removals of the device (two reports). SEARCH STRATEGY: We searched both MEDLINE and EMBASE, handsearches of journals through CENTRAL, and lists of references. We also wrote to international experts in the field to identify unpublished studies. SELECTION CRITERIA: We included randomized controlled trials using any antibiotic compared with a placebo. We found four such trials; two had pilot study data available. DATA COLLECTION AND ANALYSIS: DATA EXTRACTION: We used searches of MEDLINE, EMBASE, and handsearches of journals available through CENTRAL. We also reviewed lists of references in original research and in review articles. We wrote to experts to identify unpublished trials and made telephone calls to authors to supply missing information. Two independent reviewers abstracted data. We assessed the validity of each study using methods suggested in the Cochrane Handbook. DATA SYNTHESIS: We generated 2x2 tables for the principal outcome measures. We used the Peto modified Mantel-Haenszel technique to calculate odds ratios and assessed statistical heterogeneity between studies. MAIN RESULTS: The odds ratios for pelvic inflammatory disease associated with use of prophylactic doxycycline or azithromycin compared with placebo or no treatment was 0.89 (95%CI 0.53-1.51). Use of prophylaxis was associated with a small reduction in unscheduled vists to the provider (OR 0.82; 95% CI 0.70-0.98). Use of doxycycline or azithromycin had little effect on the likelihood of removal of the IUD within 90 days of insertion (OR 1.05; 95% CI 0.68-1.63). Significant heterogeneity did not exist between studies. REVIEWER'S CONCLUSIONS: Use of either doxycycline 200 mg or azithromycin 500 mg by mouth before IUD insertion confers little benefit. While the reduction in unscheduled visits to the provider was marginally significant, the cost-effectiveness of routine prophylaxis remains questionable. A uniform finding in these trials was the low risk of IUD associated infection, with or without use of antibiotic prophylaxis. PMID- 11405987 TI - Oral misoprostol for induction of labour. AB - BACKGROUND: Prostaglandins are hormones naturally present in the uterus that cause contractions during labour. A synthetic prostaglandin analogue misoprostol is produced in tablets that can be given orally or vaginally, but it is not yet licensed for use in pregnancy. Unlicensed use of misoprostol in pregnancy is increasingly common, because misoprostol is cheap, stable at room temperature and effective in causing uterine contractions. Oral use of the drug misoprostol may be convenient, but high doses could cause uterine hyperstimulation and uterine rupture which may be life-threatening for both mother and fetus. OBJECTIVES: The objective of this review was to assess the effects of oral misoprostol used for labour induction in women with a viable fetus in the third trimester of pregnancy. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register were searched in December 2000. SELECTION CRITERIA: Randomised trials of oral misoprostol versus other methods, placebo or no treatment, given to women with a viable fetus for labour induction. DATA COLLECTION AND ANALYSIS: This is one of a series of the Cochrane reviews of methods of cervical ripening and labour induction using standardised methodology. This review includes comparisons between oral misoprostol with placebo, vaginal prostaglandins, intracervical prostaglandins, oxytocin, amniotomy, oxytocin and amniotomy or vaginal misoprostol. Data from all relevant trials are extracted by the reviewer using centrally designed data sheets. MAIN RESULTS: One trial with 80 randomised women with prelabour rupture of membranes at term showed that, compared with placebo, oral misoprostol reduces the need for oxytocin infusion from 51 percent to 13 percent (relative risk 0.25, 95% confidence interval (CI) 0.1 to 0.6) and shortens delivery time by 8.7 hours (95% CI 6.0 to 11.3). Compared with vaginal or intracervical prostaglandins, oral misoprostol showed no beneficial or harmful effects. However, only two trials with 962 randomised women in total compared oral misoprostol with vaginal dinoprostone and one trial with 200 women compared oral misoprostol with intracervical dinoprostone. Two small trials with 188 women in total compared oral misoprostol and oxytocin in women with term ruptured membranes and found no significant differences in prespecified outcomes. In seven trials with 1278 randomised women that compared oral with vaginal misoprostol, oral misoprostol appeared to be less effective. More women in the oral misoprostol group did not achieve vaginal delivery within 24 hours of randomisation (50%) compared with 39.7% in the vaginal misoprostol group (relative risk 1.27, 95% confidence intervals 1.09 to 1.47). The caesarean section rate was lower in the oral misoprostol group (16.7%) compared with 21.7% in the vaginal misoprostol group (relative risk 0.77, 95% confidence intervals 0.61 to 0.97). There was no difference in uterine hyperstimulation with fetal heart rate changes (8.5% versus 7.4%; relative risk 1.11, 95% confidence intervals 0.78 to 1.59). There were no reported cases of severe neonatal and maternal morbidity. REVIEWER'S CONCLUSIONS: Oral misoprostol is an effective method for labour induction in the third trimester. However, the data on optimal regimens and safety are lacking. It is possible that effective oral regimens may have an unacceptably high incidence of complications such as uterine hyperstimulation and possibly uterine rupture. PMID- 11405988 TI - Antibiotics versus placebo for prevention of postoperative infection after appendicectomy. AB - BACKGROUND: Appendicitis is the most common cause of acute abdominal pain requiring surgical intervention. The cause of appendicitis is unclear and the mechanism of pathogenesis continues to be debated. Despite improved asepsis and surgical techniques, postoperative complications, such as wound infection and intraabdominal abscess, still account for a significant morbidity. Several studies implicate that postoperative infections are reduced by administration of antimicrobial regimes. OBJECTIVES: The objective of this review is to compare the use of antibiotics with placebo or no treatment in patients undergoing appendectomy. Will these patients benefit from antimicrobial prophylaxis? The outcomes are described according to the nature of the appendix, as either simple appendicitis (including the non-infectious stage) and complicated appendicitis. This review do not attempt to compare the effect of different regimens, a clinical question that is addressed in another review undertaken by this Group (CCCG). SEARCH STRATEGY: We searched The Cochrane Controlled Trials Register (Cochrane Library 2000 issue 4), Medline (January 1966 to September 2000), Embase and the Cochrane Colorectal Cancer Group specialised register (September 2000). In addition we manually searched the reference lists of the primary identified trials. SELECTION CRITERIA: Randomised Controlled Trials (RCT) and Controlled Clinical Trials (CCT) in which any antibiotic regime were compared to placebo in patients suspected of having appendicitis undergoing appendectomy were evaluated. Both studies on children and adults were reviewed. The outcome measures of the studies were either wound infection, intraabdominal abscess, length of stay in hospital or mortality. DATA COLLECTION AND ANALYSIS: Eligibility and trial quality were assessed, recorded and cross-checked by to reviewers. MAIN RESULTS: Forty-four studies including 9298 patients were included in this review. The overall result is that use of antibiotics is superior to placebo for the outcome wound infection and intraabdominal abscess, with no apparent difference in the nature of the removed appendix. Studies exclusively on children and studies examining topical application reported results in favour to the above although the results were not significant. REVIEWER'S CONCLUSIONS: Antibiotic prophylaxis is effective in the prevention of postoperative complications in appendectomised patients, whether the administration are given pre-, per- and post-operatively and could be considered for routine in emergency appendectomies. PMID- 11405989 TI - Iron therapy for improving psychomotor development and cognitive function in children under the age of three with iron deficiency anaemia. AB - BACKGROUND: Iron deficiency and iron deficiency anaemia (IDA) are common in young children. It has been suggested that the lack of iron may have important effects on children's psychomotor development and cognitive function. OBJECTIVES: To determine the effects of iron therapy on psychomotor development and cognitive function in iron deficient children less than 3 years of age. SEARCH STRATEGY: The following databases were searched: COCHRANE LIBRARY (2000 ISSUE 4) MEDLINE (1966-August 2000) EMBASE (1980-August 2000) Latin American Database (LILACS) PsycLIT Journal articles (1974-August 2000) PsycLIT Chapters and Books (1987 August 2000) The references of identified trials and of important review articles were scrutinised. Citation searches on trials from the primary search were performed within the Science Citation Index. Key authors were contacted. SELECTION CRITERIA: Studies were included if children less than 3 years of age with evidence of iron deficiency anaemia were randomly allocated to iron or iron and vitamin C versus a placebo or vitamin C alone and assessment of developmental status or cognitive function was carried out using standardised tests by observers blind to treatment allocation. DATA COLLECTION AND ANALYSIS: Abstracts and titles of studies identified on searches of electronic databases were read to determine whether they might meet the inclusion criteria. Full copies of those possibly meeting these criteria from electronic or other searches were assessed by two independent reviewers. Differences of opinion about suitability for inclusion were resolved by discussion. Data were analysed separately depending on whether participants had iron assessments were performed within one month of beginning iron therapy or later. MAIN RESULTS: Five trials, including 180 children with IDA, examined the effects of iron therapy on measures of psychomotor development between 5 and 11 days of commencement of therapy. Data from four trials could be pooled. The pooled difference in pre to post treatment change in Bayley Scale PDI between iron treated and placebo groups was -3.2 (95%CI -7.24, 0.85) and in Bayley Scale MDI, 0.55 (95% CI -2.84, 1.75). Two studies, including 160 randomised children with IDA, examined the effects of iron therapy on measures of psychomotor development more than 30 days after commencement of therapy. Aukett et al reported the mean number of skills gained after two months of iron therapy, using the Denver test. The intervention group gained 0.8 (95% CI -0.18, 1.78) more skills on average than the control group. Idjrandinata et al reported that the difference in pre to post treatment change in Bayley Scale PDI between iron treated and placebo groups after 4 months was 18.40 (95%CI 10.16, 26.64) and in Bayley Scale MDI, 18.80 (95% CI 10.19, 27.41). REVIEWER'S CONCLUSIONS: There is no convincing evidence that iron treatment of young children with IDA has an effect on psychomotor development discernable within 5-11 days. The effect of longer term treatment remains unclear but the data would be compatible with clinically significant benefit. There is urgent need for further randomised controlled trials with long term follow up. PMID- 11405990 TI - Venepuncture versus heel lance for blood sampling in term neonates. AB - BACKGROUND: Heel lance has been the conventional method of blood sampling in neonates for screening tests. Neonates undergoing this procedure experience pain. Despite various studies evaluating the role of pharmacological and non pharmacological interventions to date, there are no effective and practical methods to alleviate pain from heel lance. OBJECTIVES: To determine whether venepuncture or heel lance is less painful and more effective for blood sampling in term neonates. SEARCH STRATEGY: Systematic search in accordance with Cochrane Neonatal Collaborative Review Group. All randomized controlled trials which compared pain response to venepuncture vs. heel lance were searched using MEDLINE, EMBASE, CINAHL, the Cochrane Library, reference lists of identified trials and personal files. No language restrictions were applied. SELECTION CRITERIA: Randomized controlled trials which compared pain response to venepuncture vs. heel lance were selected for this review. DATA COLLECTION AND ANALYSIS: Data regarding the primary outcome of pain response to venepuncture vs. heel lance as assessed by validated pain measures were abstracted. Secondary outcomes including the need of repeat blood sampling, bruising/hematoma at local site, and parental perception of their own anxiety and infant's pain were also abstracted. All data were analysed using RevMan 4.1. When possible, meta-analysis was done using relative risk (RR) and event rate difference (ERD), along with their 95% confidence intervals (CI). If ERD was significant, number needed to treat (NNT) was calculated. Weighted mean difference (WMD) was used for continuous data. When present, statistically significant between study heterogeneity was reported. MAIN RESULTS: Three trials were eligible for inclusion in the review. Pain assessments were made using validated pain measures including Neonatal Infant Pain Scale (NIPS), Neonatal Facial Action Coding System (NFCS), Premature Infant Pain Profile (PIPP) score and cry characteristics. Despite the heterogeneity in the pain measures used, all studies showed statistically significantly lower pain scores for venepuncture as compared to heel lance. In one study, maternal anxiety was noted to be higher in the venepuncture group as compared to heel lance group prior to the procedure; however, after observing the procedure, mothers rated their infant's pain to be lower in the venepuncture group. A meta-analysis was performed on the need for at least one additional skin puncture to obtain the required amount of blood using venepuncture as compared to heel lance. The relative risk for requiring more than one skin puncture for venepuncture versus heel lance was 0.30 [95% confidence interval (CI) 0.18, 0.49]. The risk difference was -39% (95% CI -50%, -28%). For this outcome there was statistically significant between study heterogeneity (for RR, p=0.02; for RD, p=0.0001). The number needed to treat (NNT) to avoid one repeat skin puncture was 2.56 (95% CI 2, 3.57). REVIEWER'S CONCLUSIONS: Venepuncture, when performed by a trained phlebotomist, appears to be the method of choice for blood sampling in term neonates. For each two to three venepunctures instead of heel lance, the need for one additional skin puncture can be avoided. In view of the promising results derived from these small studies with some methodological limitations, including statistically significant between study heterogeneity, further well designed randomized controlled trials need to be conducted. The interventions should be compared in settings where several individuals perform the venepuncture and/or the heel lance. PMID- 11405991 TI - Diuretics for respiratory distress syndrome in preterm infants. AB - BACKGROUND: Lung edema may complicate respiratory distress syndrome (RDS) in preterm infants. OBJECTIVES: The aim of this review was to assess the risks and benefits of diuretic administration in preterm infants with RDS. SEARCH STRATEGY: We used the standard search method of the Cochrane Neonatal Review Group. We searched Medline, Embase and the Cochrane Controlled Trials Register from the Cochrane Library, using the following keywords: and . In addition, we searched the abstract books of the American Thoracic Society and Pediatric Research Societies. SELECTION CRITERIA: We only included trials in which preterm infants with RDS and less than 5 days of age were randomly allocated to diuretic administration. Of those trials, we only included studies in which at least one of the following outcomes measures was evaluated: mortality, patent ductus arteriosus, hypovolemic shock, intraventricular hemorrhage, renal failure, duration of oxygen supplementation, duration of mechanical ventilation, need for oxygen supplementation at 28 days of life, oxygen supplementation at 36 weeks of postconceptional age (gestational age + postnatal age), length of stay, number of rehospitalizations during the first year of life, and neurodevelopmental outcome. DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study. Any disagreement was resolved by discussion. MAIN RESULTS: Six studies met inclusion criteria. Studies available for this systematic review were all done before the current era of prenatal steroids, surfactant, indomethacin and fluid restriction. Furosemide administration had no long-term benefits. Furosemide-induced transient improvement in pulmonary function did not outweigh an increased risk for patent ductus arteriosus and for hemodynamic instability. REVIEWER'S CONCLUSIONS: There are no current data to support routine diuretic administration in preterm infants with RDS. Elective administration of furosemide or any diuretic to any patient with RDS should be carefully weighed against the risk of precipitating hypovolemia. In addition, elective administration of furosemide should be weighed against the risk of developing a symptomatic patent ductus arteriosus. PMID- 11405992 TI - Potassium nitrate toothpaste for dentine hypersensitivity. AB - BACKGROUND: Dentine hypersensitivity may be defined as the pain arising from exposed dentine, typically in response to external stimuli, and which cannot be explained by any other form of dental disease. Many treatment regimes have been recommended over the years, and in recent years particular attention has been focused on toothpastes containing potassium nitrate. OBJECTIVES: To compare the effectiveness of potassium nitrate containing toothpastes with placebo toothpastes in reducing dentine hypersensitivity. SEARCH STRATEGY: The following databases were cross searched via the database host DIALOG: MEDLINE, EMBASE, ELSEVIER BIOBASE, BIOSIS PREVIEWS, CAB HEALTH, SCI SEARCH, CURRENT CONTENTS until 1 April 2000. The specialised Cochrane Oral Health Group Trials Register was also searched. Bibliographies of clinical studies and reviews identified in the electronic search were checked for studies published outside the electronically searched journals. SELECTION CRITERIA: Randomised clinical trials (RCTs) in which the effect on dentine hypersensitivity of potassium nitrate toothpastes were tested against non-potassium nitrate containing placebo toothpastes. DATA COLLECTION AND ANALYSIS: Two of the reviewers independently recorded the results of the included trials using a specially designed chart. Sensitivity was assessed by using thermal, tactile, air blast, and subjective methods. The quality of all RCTs, that fulfilled the inclusion criteria, was acceptable with Jadad scores ranging from 3 to 4 (Jadad 1998). MAIN RESULTS: Out of the eight studies that initially fulfilled the criteria to be included in the review, four studies did not present mean and standard deviations and could thus not be included in the meta-analysis. Three of these did not show an effect on any of the measurements of dentine hypersensitivity, while one did. Four studies were included in the meta-analysis which showed statistically significant effect of potassium nitrate toothpaste on air blast and tactile sensitivity, e.g. the meta analysis of air blast sensitivity showed a standardized mean difference in sensitivity score of 1.51 (95% CI: -2.09 to -0.94) in favour of treatment. The subjective assessment failed to show a significant effect at the six to eight week assessment. REVIEWER'S CONCLUSIONS: No strong evidence is available supporting the efficacy of potassium nitrate toothpaste for dentine hypersensitivity. PMID- 11405993 TI - Vitamin A for treating measles in children. AB - BACKGROUND: Measles is a leading cause of childhood morbidity and mortality. Vitamin A deficiency is a recognised risk factor for severe measles. The World Health Organization (WHO) recommends administration of an oral dose of 200,000 IU (or 100,000 IU in infants) of vitamin A per day for two days to children with measles in areas where vitamin A deficiency may be present. OBJECTIVES: The purpose of this review is to determine whether vitamin A when commenced after measles has been diagnosed, is beneficial in preventing mortality, pneumonia and other complications in children. SEARCH STRATEGY: MEDLINE and the Cochrane Library, Issue 4, 1999 were searched. SELECTION CRITERIA: Only randomized controlled trials in which children with measles were given vitamin A or placebo along with standard treatment were considered. DATA COLLECTION AND ANALYSIS: Studies were assessed independently by two reviewers. The analysis of dichotomous outcomes was done using the StatExact software package. Sub-group analyses were done for dose, formulation, age, hospitalisation and pneumonia specific mortality. Weighted mean difference with 95% CI were calculated for continuous outcomes. MAIN RESULTS: The relative risks (RR) and 95% Confidence Intervals (CI) are based on the estimates from the StatExact software package. There was no significant reduction in mortality in the vitamin A group when all the studies were pooled together (RR 0.60; 95% CI 0.32 to 1.12)(Statexact estimate). There was a 64% reduction in the risk of mortality in children who were given two doses of 200,000 IU of vitamin A (RR=0.36; 95% CI 0.14 to 0.82) as compared to placebo. Two doses of water based vitamin A were associated with a 81% reduction in risk of mortality (RR=0.19; 95% CI 0.02 to 0.85) as compared to 48% seen in two doses of oil based preparation (RR=0.52; 95% CI 0.16 to 1.40). Two doses of oil and water based vitamin A were associated with a 82% reduction in the risk of mortality in children under the age of 2 years (RR=0.18; 95% CI 0.03 to 0.61) and a 67% reduction in the risk of pneumonia specific mortality (RR=0.33; 95% CI 0.08 to 0.92). There was no evidence that vitamin A in a single dose of 200,000 IU was associated with a reduced risk of mortality among children with measles (RR=0.77; 95% CI 0.34 to 1.78). Sub-groups like age, dose, formulation, hospitalisation and case fatality in the study area were highly correlated and there were not enough studies to separate out the individual effects of these factors. There was a 47% reduction in the incidence of croup (RR=0.53; 95% CI 0.29 to 0.89), while there was no significant reduction in the incidence of pneumonia (RR=0.92; 95% CI 0.69 to 1.22) or of diarrhoea (RR=0.80; 95% CI 0.27 to 2.34). Duration of diarrhoea was measured in days and there was a reduction in its duration of almost two days WMD -1.92, 95% CI -3.40 to -0.44. Only one study evaluated otitis media and found a 74% reduction in its incidence (RR=0.26, 95% CI, 0.05 to 0.92). We did not find evidence that a single dose of 200,000 IU of vitamin A per day, given in oil based formulation in areas with low case fatality, was associated with reduced mortality among children with measles. However, there was evidence that the same dose given for two days was associated with a reduced risk of overall mortality and pneumonia specific mortality. REVIEWER'S CONCLUSIONS: Although we did not find evidence that a single dose of 200,000 IU of vitamin A per day was associated with reduced mortality among children with measles, there was evidence that the same dose given for two days was associated with a reduced risk of overall mortality and pneumonia specific mortality. The effect was greater in children under the age of two years. There were no trials that compared a single dose with two doses, although the precision of the estimates of trials that used a single dose were similar to the trials that used two doses. PMID- 11405994 TI - Holding chambers versus nebulisers for inhaled steroids in chronic asthma. AB - BACKGROUND: Inhaled corticosteroids are available in the form of a suspension for nebulisation, although the role of this mode of therapy in the treatment of chronic asthma is still unclear. OBJECTIVES: To assess the efficacy and safety of inhaled corticosteroids delivered via nebuliser versus holding chamber for the treatment of chronic asthma. SEARCH STRATEGY: We searched the Cochrane Airways Group Trial Register (1999) and reference lists of articles. We contacted the authors of studies and pharmaceutical companies for additional studies and hand searched the British Journal of Clinical Research, European Journal of Clinical Research and major respiratory society meeting abstracts (1997-1999). SELECTION CRITERIA: Randomised controlled trials comparing nebuliser to holding chamber in the delivery of inhaled corticosteroids for the treatment of chronic asthma. All age groups of patients were considered. Two reviewers assessed articles for inclusion; two reviewers independently assessed included studies for methodological quality. DATA COLLECTION AND ANALYSIS: One reviewer extracted data; authors were contacted to clarify missing information. Quantitative analyses were undertaken using Review Manager 4.1 with MetaView 3.1. MAIN RESULTS: Two studies were selected for inclusion (63 subjects), both concerned adults. Methodological quality was variable. Due to design differences it was not appropriate to pool the studies. The single high quality study compared budesonide 2000-8000 mcg delivered via Pari Inhalier Boy jet nebuliser with inspiration-only inhalation to budesonide 1600 mcg via large volume spacer. The nebuliser delivery led to higher morning peak expiratory flow values (25 L/min p<0.01), higher evening values (30L/min, p<0.01), lower rescue beta2 agonist use and symptom scores compared to the holding chamber delivery. REVIEWER'S CONCLUSIONS: Budesonide in high dose delivered by the particular nebuliser used in the only double-blinded study that could be included in this review was more effective than budesonide 1600 mcg via a large volume spacer. However, it is not clear whether this was an effect of nominal dose delivered or delivery system. Cost, compliance and patient preference are important determinants of clinical effectiveness that have not been assessed. Future studies are needed to evaluate the relative effectiveness of inhaled corticosteroids delivered by different combinations of nebuliser/compressor compared to holding chamber. Moreover, further studies assessing these delivery methods are needed in infants and pre school children, as these are groups that are likely to be considered for treatment with nebulised corticosteroids. PMID- 11405995 TI - Thiamine for Alzheimer's disease. AB - BACKGROUND: Vitamin B1 (thiamine) plays an important role in Wernicke-Korsakoff syndrome (a form of amnesia caused by brain damage occurring in long-term alcoholics who rely mainly on alcohol for nutrition). The acute syndrome is normally reversible but may proceed to profound dementia, although its progress can be stopped by a timely injection of a large dose of thiamine. There have been suggestions that thiamine may have a beneficial effect in Alzheimer's disease. OBJECTIVES: The objective of this systematic review is to evaluate the efficacy of thiamine for people with Alzheimer's disease. SEARCH STRATEGY: The Cochrane Controlled Trials Register (Issue 3:2000), the CDCIG Trials Register and other sources were searched for this update in July 2000 using the terms 'alzheimer*', thiamin* and vitamin B1'. In addition bibliographies of published reviews, and conference proceedings were searched and pharmaceutical companies and trials investigators were contacted. SELECTION CRITERIA: All unconfounded, double-blind, randomized trials in which treatment with thiamine was administered for more than a day and compared with placebo in patients with dementia of the Alzheimer's type. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers and the odds ratios (95% CI) or the average differences (95% CI) were estimated. MAIN RESULTS: There are three included studies, but few results were reported that could be included. The cross-over studies did not report results from the first phase. It was not possible to pool any results for a meta analysis. Nolan 1991 reports results that show no evidence of an effect on MMSE at 3, 6, 9 and 12 months for thiamine compared with placebo for those who completed the trial. Meador 1993a noted that 3/8 on thiamine compared with 6/9 on placebo were worse as measured on the ADAS-Cog at 3 months compared with baseline, but the difference is not statistically significant. Blass 1988 and Nolan 1991 reported that no significant side-effects were noted during the study, and Meador 1993a did not mention side-effects. Blass 1988 noted that 5/16 and Nolan 1991 that 5/15 did not complete the study, but neither mentioned the groups to which these people belonged. REVIEWER'S CONCLUSIONS: It is not possible to draw any conclusions from this review. The number of people included in the studies if less than 50 and the reported results are inadequate. PMID- 11405996 TI - Physostigmine for Alzheimer's disease. AB - BACKGROUND: The main pharmacological approach for the treatment of Alzheimer's disease (AD) has been based on the use of agents potentiating cholinergic transmission, particularly by inhibiting acetylcholinesterase (AChE), the enzyme that destroys acetylcholine after it has been secreted into the synaptic clefts. Physostigmine is an AChE inhibitor originally extracted from calabar beans. It is licensed in many countries as an agent for reversing the effect of drugs and poisons causing the anticholinergic syndrome. Studies conducted more than 20 years ago suggested that physostigmine could improve memory in people with or without dementia. Investigation of this property has been limited by the very short half-life of physostigmine. Various forms of administering the drug have been tried to overcome this problem, most recently a controlled-release (CR) oral formulation, and a skin patch. It has been proposed as a potential drug for the symptomatic treatment of AD. OBJECTIVES: To determine whether there is evidence of beneficial effects for the use of physostigmine in Alzheimer's disease. To assess the incidence and severity of adverse effects. SEARCH STRATEGY: The Cochrane Controlled Trials Register was searched using the following terms: 'physostigmine', 'physostigmine salicylate', 'Synapton' and 'Antilirium' in accordance with the Cochrane Dementia and Cognitive Improvement Group's search strategy. The pharmaceutical company was contacted. SELECTION CRITERIA: All relevant unconfounded, double-blind, randomized, placebo-controlled trials in which physostigmine was administered for more than one day to patients with dementia of Alzheimer type. Trials in which the allocation to the treatment was not randomized, or in which the allocation to the treatment was not concealed were excluded. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers (JMC & JB), pooled where appropriate and possible, and the weighted or standardized mean differences or Peto odds ratios (95% CI) were estimated. Where possible, intention-to-treat analysis was used. MAIN RESULTS: Fifteen studies were included using four different methods of administration of physostigmine. Four studies, involving 29 people in total, used intravenous infusion; seven, involving 131 people, used a conventional oral form; four, involving 1456 participants, used a controlled-release oral form, and one study of 181 people used a verum skin patch. There are no usable results from the intravenous infusion trials, and the few results from the conventional oral form showed no benefit of physostigmine compared with placebo. The results from two of the four studies of the controlled-release physostigmine apply only to a group of patients identified as responders in a pre-randomization titration period. The best dose physostigmine (mean 25mg/day) was associated with a 1.75 point improvement on ADAS-Cog score (mean difference -1.75, 95% confidence interval 2.90, -0.60 on an intention-to-treat basis) and a 0.26 point improvement on the CGIC score (treated as a continuous scale) (mean difference -0.26, 95% confidence interval 0.06, 0.46 on an intention-to-treat basis) compared with placebo at 6 weeks. There were statistically significantly higher numbers of patients from the physostigmine group withdrawing from the trial (22/183 vs 2/183)(OR 5.92, 95% confidence limits 2.59, 13.54) and suffering at least one event of nausea, vomiting, diarhoea, anorexia, dizziness, stomach pain, flatulence or sweating compared with placebo at 6 weeks. The best dose physostigmine (mean 27mg/day) was associated with a 2.0 point improvement on ADAS-Cog score (mean difference -2.02, 95% confidence interval -3.59, -0.45 on an intention to treat basis) compared with placebo at 12 weeks. There were statistically significantly higher numbers of patients from the physostigmine group withdrawing from the trial due to adverse events (13/83 vs 5/93)(OR 3.05, 95% confidence limits 1.15, 8.07) and suffering at least one event of nausea, vomiting, diarhoea, anorexia, dizziness, stomach pain, tremor, asthenia or sweating compared with placebo at 12 weeks. When no attempt was made to identify responders and all relevant patients with Alzheimer's disease were randomized, fixed dose physostigmine (mean 33 mg/day) was associated with a statistically significantly higher number withdrawing (234/358 vs 31/117)(OR 4.82, 95% confidence limits 3.17, 7.33), withdrawing due to adverse events (196/358 vs 10/117) (OR 6.54, 95%confidence limits 4.29, 9.95) and suffering at least one event of nausea, vomiting, diarhoea, anorexia, dizziness, stomach pain, dyspepsia, sweating, asthenia, dyspnoea or abnormal dreaming compared with placebo at 24 weeks. The results from the study of the verum patch physostigmine show that the double dose (delivering mean dose 12mg/day) was associated with statistically significantly higher numbers suffering at least one adverse event of vomiting, nausea or abdominal cramps compared with placebo at 24 weeks, but placebo was associated with statistically significantly greater numbers of gastrointestinal complaints at 24 weeks compared with single-dose physostigmine. REVIEWERS' CONCLUSIONS: The evidence of effectiveness of physostigmine for the symptomatic treatment of Alzheimer's disease is limited. Even in a controlled release formulation designed to overcome the short half-life, physostigmine showed no convincing benefit and adverse effects remained common leading to a high rate of withdrawal. PMID- 11405997 TI - Corticosteroid therapy for nephrotic syndrome in children. AB - BACKGROUND: In nephrotic syndrome protein leaks from the blood to the urine through the glomeruli resulting in hypoproteinaemia and generalised oedema. Children with untreated nephrotic syndrome frequently die from infections. The majority of children with nephrotic syndrome respond to corticosteroids. However about 70% of children experience a relapsing course with recurrent episodes of oedema and proteinuria. Corticosteroid usage has reduced the mortality rate in childhood nephrotic syndrome to around 3%, with infection remaining the most important cause of death. However corticosteroids have known adverse effects such as obesity, poor growth, hypertension, diabetes mellitus, osteoporosis and adrenal suppression. The original treatment schedules for childhood nephrotic syndrome were developed in an ad hoc manner. The optimal doses and durations of corticosteroid therapy that are most beneficial and least harmful have not been clarified. The aim of this systematic review was to assess the benefits and harms of corticosteroid therapy in treating children with nephrotic syndrome. OBJECTIVES: To determine the benefits and harms of different corticosteroid regimens in preventing relapse in children with steroid responsive nephrotic syndrome (SRNS). SEARCH STRATEGY: Published and unpublished randomised controlled trials were identified from the Cochrane Controlled Trials Register, Medline, Embase, reference lists of articles, abstracts from proceedings and contact with known investigators in the area. SELECTION CRITERIA: Randomised trials were included if they were carried out in children (aged three months to 18 years) in their initial or subsequent episode of SRNS, if they compared different durations, total doses or other dose strategies using prednisone or other corticosteroid agent and if they had outcome data at six months or more. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed all eligible studies for inclusion, assessed study quality and extracted data. The principle outcome measure was the number of children with and without relapse after six and 12-24 months. Secondary outcomes sought included the number of children who developed frequently relapsing nephrotic syndrome and adverse events. A random effects model was used to estimate summary effect measures (relative risk RR, risk difference RD) after testing for heterogeneity. Meta-regression was used to explore potential between-study differences due to the baseline risk of relapse, study quality and types of interventions used. MAIN RESULTS: Twelve trials were identified. A meta-analysis of five trials, which compared two months of prednisone with three months or more in the first episode, showed that the longer duration significantly reduced the risk of relapse at 12 - 24 months (relative risk 0.73; 95% CI 0.60,0.89) without an increase in adverse events. There was an inverse linear relationship (RR = 1.382 (SE 0.215) - 0.133 duration (SE 0.048); r2 = 0.66; p = 0.05) between the duration of treatment and risk of relapse. The number of children who became frequent relapsers and the mean relapse rate/patient/year were also significantly reduced without increase in serious adverse events. In children with frequently relapsing nephrotic syndrome, deflazacort was significantly more effective in maintaining remission than prednisone (RR 0.44; 95% CI 0.25, 0.78). REVIEWER'S CONCLUSIONS: From this meta analysis of randomised controlled trials it can be concluded that children in their first episode of nephrotic syndrome should be treated for at least three months with an increase in benefit being demonstrated for up to seven months of treatment. In a population with a baseline risk for relapse following the first episode of 60% with two months of prednisone, daily prednisone for four weeks followed by alternate day therapy for six months would be expected to reduce the number of children experiencing a relapse by about 40%. In children who relapse frequently, deflazacort deserves further study. PMID- 11405998 TI - Aerosolized diuretics for preterm infants with (or developing) chronic lung disease. AB - BACKGROUND: Lung disease in preterm infants is often complicated with lung edema. OBJECTIVES: The aim of this review is to assess the risks and benefits of aerosolized diuretic administration in preterm infants with or developing chronic lung disease (CLD). Primary objectives are to assess effects on short term outcome (changes in need for oxygen or ventilatory support) and effects on long term outcome. Secondary objectives are to assess changes in pulmonary mechanics and potential complications of therapy. SEARCH STRATEGY: We used the standard search method of the Cochrane Neonatal Review Group. We used the following keywords: [ or ] and , limited to and limited to or . We searched Medline (1966-1998), Embase (1974-1998) and the Cochrane Controlled Trials Register (CCTR) from the Cochrane Library (1998, Issue 4). In addition, we hand searched several abstract books of national and international American and European Societies. The search of MEDLINE was updated in January, 2001. SELECTION CRITERIA: We included in this analysis trials in which preterm infants with or developing chronic lung disease and at least five days of age were all randomly allocated to receive an aerosolized loop diuretic. Eligible studies needed to assess at least one of the outcome variables defined a priori for this systematic review. Primary outcome variables included important clinical outcomes, and secondary outcome variables included pulmonary mechanics and potential complications of therapy. DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration which is described in the Cochrane Collaboration Handbook. Two investigators extracted, assessed and coded separately all data for each study, using a form that was designed specifically for this review. Any disagreement was resolved by discussion. We combined parallel and cross-over trials and, whenever possible, transformed baseline and final outcome data measured on a continuous scale into change scores using Follmann's formula. MAIN RESULTS: We identified eight studies which met selection criteria. Most studies focused on pathophysiological parameters and did not assess effects on important clinical outcomes defined in this review or the potential complications of diuretic therapy. No study assessed the amount of diuretic effectively delivered to the patient. Furosemide was the only diuretic used in the eight studies included in this review. Among preterm infants < 3 weeks of age developing CLD, not enough information is available to assess the effect of aerosolized furosemide on outcome or lung function. Among infants > 3 weeks with CLD, a single aerosolized dose of 1 mg/kg of furosemide may transiently improve pulmonary mechanics. Not enough information is available to assess the effect of chronic administration of aerosolized furosemide on oxygenation and pulmonary mechanics. REVIEWER'S CONCLUSIONS: In preterm infants > 3 weeks with CLD administration of a single dose of aerosolized furosemide improves pulmonary mechanics. In view of the lack of data from randomized trials concerning effects on important clinical outcomes, routine or sustained use of aerosolized loop diuretics in infants with (or developing) CLD cannot be recommended based on current evidence. More double-blinded randomized trials are needed (1) to analyze factors likely to affect the response to aerosolized furosemide, e.g., washout period and delivery of furosemide to distal airways, and (2) to assess the effects of chronic administration of aerosolized furosemide on mortality, O2 dependency, ventilator dependency, length of hospital stay and long-term outcome. PMID- 11405999 TI - Timing of surgery for aneurysmal subarachnoid haemorrhage. AB - BACKGROUND: The timing of surgery to secure a ruptured aneurysm after a subarachnoid haemorrhage is an important issue. Early clipping of an aneurysm prevents rebleeding, a major cause of death after a subarachnoid haemorrhage. However, concerns about the possible deleterious effects of early surgery raise questions about the safety and efficacy of this approach. This review examines the randomised controlled evidence addressing the effect of surgery at different time intervals on the outcome after a subarachnoid haemorrhage. OBJECTIVES: To determine whether the timing of surgery after a subarachnoid haemorrhage significantly influences overall management outcome. SEARCH STRATEGY: We searched the Cochrane Stroke Review Group Trials Register and in addition searched MEDLINE, EMBASE and the Cochrane Controlled Trials Register (CENTRAL/CCTR). Colleagues were contacted to identify further studies and unpublished trials. SELECTION CRITERIA: All completed, unconfounded, truly randomised trials comparing "best medical treatment plus early surgery" with "best medical treatment plus delayed surgery". DATA COLLECTION AND ANALYSIS: The authors selected trials for inclusion, or exclusion, according to the above criteria. An "intention to treat" analysis strategy was utilised. MAIN RESULTS: Only one randomised controlled trial addressing the timing of surgery after aneurysmal subarachnoid haemorrhage was identified. Patients undergoing early surgery tended to fare better than those undergoing late surgery (death or dependency at 3 months OR 0.37 95% CI 0.13,1.02). Patients undergoing surgery in the intermediate time period appeared to fare worse than those undergoing early surgery although confidence intervals were wide (death or dependency at 3 months OR 0.34 95% CI 0.12, 0.93). REVIEWER'S CONCLUSIONS: Based upon the limited randomised controlled evidence available, the timing of surgery was not a critical factor in determining outcome following a subarachnoid haemorrhage. Since the publication of the only randomised controlled study in 1989, techniques for the treatment of subarachnoid haemorrhage have progressed, questioning the validity of the conclusions in the modern era. Currently, most neurovascular surgeons elect to operate within 3 or 4 days of the bleed in good grade patients to minimise the chances of a devastating rebleed. However, the treatment of patients in poorer grades warrants further scrutiny in a randomised controlled trial. PMID- 11406000 TI - Day centres for severe mental illness. AB - BACKGROUND: The number of people with severe mental illness who receive treatment whilst living at home has increased greatly over the last 30 years. Day centres and day hospitals frequently supplement this treatment. OBJECTIVES: To determine the effects of non-medical day centre care for people with severe mental illness. SEARCH STRATEGY: The Allied and Complementary Medicine Database (1985-1999), The British Nursing Index (1994-1998), The Cochrane Library (Issue 2, 1999), The Cochrane Schizophrenia Group's Register (May 1999), EMBASE (1980-1999), MEDLINE (1966-1999), PsycLIT (1887-1999), The Royal College of Nurses Database (1985 1996), and Sociological Abstracts (1963-1999) were searched. References of all identified studies were also inspected for more studies. SELECTION CRITERIA: All randomised controlled trials where seriously mentally ill people were allocated to non-medical day centre care. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality rated and data extracted. For dichotomous data, it had been hoped to estimate the Peto odds ratios (OR) with 95% confidence intervals (CI) and the number needed to treat statistic (NNT). Analysis was to have been by intention-to-treat. Normal continuous data were to have been summated using the weighted mean difference (WMD) and scale data presented only for those tools that had attained pre-specified levels of quality. MAIN RESULTS: Despite extensive searching, no trials were found of non-medical day centres. The electronic search identified over 300 citations but none was relevant to this review. REVIEWER'S CONCLUSIONS: The reviewers feel that the inclusion of any studies less rigorous than randomised trials would result in misleading findings and that it is not unreasonable to expect well designed, conducted and reported randomised controlled trials of day centre care. More precise nomenclature would greatly help identify relevant work. At present non-randomised comparative studies give conflicting messages about the roles provided by day centres and the clinical and social needs they are able to meet. It is therefore probably best that people with serious mental illness and their carers, if given the choice, take a pragmatic decision on which type of unit best meets their needs. There is a clear need for randomised controlled trials of day centre care compared to other forms of day care, such as day hospitals. PMID- 11406001 TI - Guided tissue regeneration for periodontal infra-bony defects. AB - BACKGROUND: Conventional treatment of destructive periodontal (gum) disease arrests the disease but does not regain the bone support or connective tissue lost in the disease process. Guided tissue regeneration (GTR) is a surgical procedure that aims to regenerate the periodontal tissues when the disease is advanced and could overcome some of the limitations of conventional therapy. OBJECTIVES: To assess the efficacy of GTR in the treatment of periodontal infra bony defects measured against the current standard of surgical periodontal treatment, open flap debridement. SEARCH STRATEGY: We conducted an electronic search of the Cochrane Oral Health Group specialised trials register and MEDLINE up to October 2000. Hand searching included Journal of Periodontology, Journal of Clinical Periodontology, Journal of Periodontal Research and bibliographies of all relevant papers and review articles up to October 2000. In addition, we contacted experts/groups/companies involved in surgical research to find other trials or unpublished material or to clarify ambiguous or missing data and posted requests for data on two periodontal electronic discussion groups. SELECTION CRITERIA: Randomised, controlled trials of at least 12 months duration comparing guided tissue regeneration (with or without graft materials) with open flap debridement for the treatment of periodontal infra-bony defects. Furcation involvements and studies specifically treating early onset diseases were excluded. DATA COLLECTION AND ANALYSIS: Screening of possible studies was conducted independently by two reviewers (RT & IN) and data abstraction by three reviewers (RT, IN & EGL). The methodological quality of studies was assessed in duplicate (RT & IN) using both individual components and a quality scale (Jadad 1998) and agreement determined by Kappa scores. Methodological quality was used in sensitivity analyses to test the robustness of the conclusions. The Cochrane Oral Health Group statistical guidelines were followed (HW) and the results expressed as weighted mean differences (WMD and 95% CI) for continuous outcomes and relative risk (RR and 95% CI) for dichotomous outcomes calculated using random effects models where significant heterogeneity was detected (P < 0.1). The final analysis was conducted using STATA 6 in order to combine both parallel group studies and intra-individual (split-mouth) studies. The primary outcome measure was gain in clinical attachment. Any heterogeneity was investigated. MAIN RESULTS: We initially included 23 trial reports. Twelve were subsequently excluded. Of these, seven presented six-months data only, three were not fully randomised controlled trials, one used a non-comparable radiographic technique. Eleven studies were finally included in the review, ten testing GTR alone and two testing GTR+bone substitutes (one study had both test treatment arms). For attachment level change, the weighted mean difference between GTR alone and open flap debridement was 1.11 mm (95% CI: 0.63 to 1.59), chi-square for heterogeneity 31.4 (df = 9), p<0.001) and for GTR+bone substitutes was 1.25 mm (95% CI: 0.89 to 1.61, chi-square for heterogeneity 0.01 (df = 1), p=0.91). GTR showed a significant benefit when comparing the numbers of sites failing to gain 2 mm attachment, with relative risk 0.58 (95% CI: 0.38, 0.88, chi-square for heterogeneity 5.72 (df = 3), p=0.13). The number needed to treat (NNT) for GTR to achieve one extra site gaining 2 mm or more attachment over open flap debridement was 8 (95% CI: 4, 33), based on an incidence of 32% of sites in the control group failing to gain 2 mm or more of attachment. For baseline incidences in the range of the control groups of 10% and 55% the NNTs are 24 and 3. Probing depth reduction demonstrated a small but statistically significant benefit for GTR, weighted mean difference 0.80 mm (95% CI: 0.14,1.46, chi-square for heterogeneity 10.0 (df = 4), p=0.04) or GTR+bone substitutes, weighted mean difference 1.24 mm (95% CI: 0.89, 1.59, chi-square for heterogeneity 0.03 (df = 1), p=0.85). No significant difference was noted for gingival recession between GTR and open flap debridement. Regarding hard tissue probing at surgical re-entry, a statistically significant greater gain was found for GTR compared with open flap debridement. This amounted to a weighted mean difference of 1.39 mm (95% CI: 1.08, 1.71, chi square for heterogeneity 0.85 (df = 2), p=0.65). For GTR+bone substitutes the difference was greater, with mean difference 3.37 mm (95% CI: 3.14, 3.61). Heterogeneity between studies was highly statistically significant for all principal comparisons and could not be explained satisfactorily by sensitivity analyses. The quality of study reporting was poor with seven out of 11 studies graded as poor using the Jadad score. (ABSTRACT TRUNCATED) PMID- 11406002 TI - Decongestants and antihistamines for acute otitis media in children. AB - BACKGROUND: Acute otitis media (AOM) is a common and important source of morbidity in children, although most cases resolve spontaneously. While frequently recommended, decongestant and antihistamine therapy is of unclear benefit. OBJECTIVES: To determine the efficacy of decongestant and antihistamine therapy in children with AOM on outcomes of AOM resolution, medication side effects, and complications of AOM. SEARCH STRATEGY: Comprehensive search of Cochrane's Controlled Trials Registry, Medline and Embase was conducted. Bibliographic review and requests for information from study authors and pharmaceutical companies supplemented this. SELECTION CRITERIA: Randomized controlled trials evaluating decongestant (DC) or antihistamine (AH) treatment for children with AOM were included. Patient-oriented outcomes were considered most relevant. There were no quality or language restrictions. DATA COLLECTION AND ANALYSIS: Investigators independently evaluated studies for inclusion, performed validity assessments, and completed data extraction. Dichotomous data were pooled to generate relative risks and numbers needed to treat, and homogeneity was assessed using approximate chi-square tests. MAIN RESULTS: For the combined control groups, healing rates at 2 weeks were high, with rates of persistent AOM <23%. No additional benefit was demonstrated from intervention subgroupings DC, AH, or any medication (DC and/or AH). Only the combined treatment (DC + AH) group demonstrated statistically lower rates of persistent AOM at the 2-week period (RR 0.76, 95% CI, NNT 10.5, 95% CI). No benefit was found for other outcomes including early or late cure rates, symptom resolution, prevention of surgery or other complications. There was an increased risk of medication side effects for those receiving an intervention, which reached statistical significance for the "any medication" and decongestant groupings (NNH 16.6, 14.3 respectively, 95% CI ). Validity subanalyses demonstrated that lower quality studies found benefit, but analysis of those studies with higher validity scores found no benefit to treatment. REVIEWER'S CONCLUSIONS: Given lack of benefit and increased risk of side effects, these data do not support the use of decongestant, antihistamine, or combined DC/AH treatment in children with AOM. The small statistical benefit found in the combination medication group is of small clinical significance and study design may be biasing the results. PMID- 11406003 TI - Antibiotic prophylaxis for mammalian bites. AB - BACKGROUND: Bites by mammals are a common problem and they account for up to 1% of all visits to hospital emergency rooms. Dog and cat bites are the most common and people are usually bitten by their own pets or by an animal known to them. School-age children make up almost a half of those bitten. Prevention of tetanus, rabies and wound infection are the priorities for staff in emergency rooms. The use of antibiotics may be useful to reduce the risk of developing a wound infection. OBJECTIVES: To determine if the use of prophylactic antibiotics in mammalian bites is effective in preventing bite wound infection. SEARCH STRATEGY: Relevant RCTs were identified by electronic searches of MEDLINE, EMBASE, LILACS and the Cochrane Controlled Trials Register databases in November 2000. SELECTION CRITERIA: We included randomised controlled trials which studied patients with bites from all mammals. Comparisons were made between antibiotics and placebo or no intervention. The outcome of interest was the number of infections at the site of bite. DATA COLLECTION AND ANALYSIS: Two reviewers extracted the data independently. All analyses were performed according to the intention-to-treat method. MAIN RESULTS: Eight studies were included. The use of prophylactic antibiotics was associated with a statistically significant reduction in the rate of infection after bites by humans. Prophylactic antibiotics did not appear to reduce the rate of infection after bites by cats or dogs. Wound type, e.g. laceration or puncture, did not appear to influence the effectiveness of the prophylactic antibiotic. Prophylactic antibiotics were associated with a statistically significant reduction in the rate of infection in hand bites (OR 0.10, 95% CI 0.01 to 0.86; NNT = 4, 95% CI 2 to 50). REVIEWER'S CONCLUSIONS: There is evidence from one trial that prophylactic antibiotics reduces the risk of infection after human bites but confirmatory research is required. There is no evidence that the use of prophylactic antibiotics is effective for cat or dog bites. There is evidence that the use of antibiotic prophylactic after bites of the hand reduces infection but confirmatory research is required. PMID- 11406004 TI - Speleotherapy for asthma. AB - BACKGROUND: Speleotherapy, the use of subterranean environments, is a therapeutic measure in the treatment of chronic obstructive airways diseases. It is virtually unknown in the UK or the US, but has considerable widespread use in some Central and Eastern European countries. OBJECTIVES: To review evidence for the efficacy of speleotherapy in the treatment of asthma. SEARCH STRATEGY: We searched electronic databases (Medline, Embase, Cochrane Airways group database), contacted speleotherapy centres and experts in the field, hand searched proceedings, and checked bibliographies of articles obtained to identify possible relevant publications. SELECTION CRITERIA: We included controlled clinical trials (i.e., both randomized and those not reporting the method of allocation) that compared clinical effects of speleotherapy with another intervention or no intervention in patients with chronic asthma. DATA COLLECTION AND ANALYSIS: Information concerning patients, interventions, results, and methodology were extracted in standardized manner by two independent reviewers and summarized descriptively. MAIN RESULTS: Three trials including a total of 124 asthmatic children met the inclusion criteria, but only one trial had reasonable methodological quality. Two trials reported that speleotherapy had a beneficial short-term effect on lung function. Other outcomes could not be assessed in a reliable manner. A further search was conducted in July 2000. One further paper was excluded (see excluded studies) REVIEWER'S CONCLUSIONS: The available evidence does not permit a reliable conclusion as to whether speleo-therapeutic interventions are effective for the treatment of chronic asthma. Randomized controlled trials with long-term follow-up are necessary. PMID- 11406005 TI - Efficacy and safety of nicotine on Alzheimer's disease patients. AB - BACKGROUND: Nicotine is a cholinergic agonist that also has a presynaptic effect in releasing acetylcholine. In animal model has been shown to reverse spatial memory deficits produced by lesions in the medial septal nucleus of rats, and in aged monkeys nicotine administration improves memory and alertness to visual stimuli. Observational studies have claimed a protective effect of smoking against Alzheimer's disease (AD), but recent studies have called this into question. Smoking is a risk factor for stroke and so, possibly, for vascular dementia. Because nicotine has adverse effects, it is important to conduct a systematic review to assess the clinical efficacy and safety of nicotine for patients with AD OBJECTIVES: To evaluate the efficacy and safety of nicotine, administered in any way or form, for people with Alzheimer's disease. SEARCH STRATEGY: The trials were identified from a search of the Specialized Register of the Cochrane Dementia and Cognitive Improvement Group on 24 January 2001 using the term nicotine. SELECTION CRITERIA: All unconfounded, double-blind, randomized trials in which treatment with nicotine patches or administration of nicotine intravenously or in any other way or form was administered for more than a day and compared with placebo for people with Alzheimer's disease. DATA COLLECTION AND ANALYSIS: The one included trial did not present results suitable for inclusion in the review. MAIN RESULTS: The poor quality of trials did not allow any synthesis of data across studies. REVIEWER'S CONCLUSIONS: This review is not able to provide any evidence that nicotine is a useful treatment for Alzheimer's disease. PMID- 11406006 TI - Interventions for vaginismus. AB - BACKGROUND: Vaginismus is involuntary vaginal muscle contraction which makes sexual intercourse difficult or impossible. It is one of the more common female psychosexual problems. Various therapeutic strategies for vaginismus, such as sex therapy and desensitization, have been considered in uncontrolled studies. OBJECTIVES: The aim of this review is to determine the clinical effectiveness of treatments for vaginismus and also to examine the role of partner participation in the effectiveness of the treatment. SEARCH STRATEGY: The Cochrane Collaboration Depression, Anxiety & Neurosis Controlled Trials Register (Issue 3, 2000), the Cochrane Controlled Trials Register (Cochrane Library, Issue 2, 2000), MEDLINE (1966 to Apr 2000), EMBASE (1980 to Apr 2000), PsycINFO (1967 to April 2000) and CINAHL (1982 to Mar 2000) were searched. The Journal of Sex Research (1974 to 1999), Sexual & Marital Therapy (1986 to 1999), Sexual Dysfunction (1998 to 1999) and the Journal of Sex and Marital Therapy (1974 to 1999) were handsearched. Reference lists and conference abstracts were searched. Experts in the field were contacted regarding unpublished material. SELECTION CRITERIA: Controlled trials comparing treatments for vaginismus with another treatment, a placebo treatment, treatment as usual or waiting list control. DATA COLLECTION AND ANALYSIS: The reviewers extracted data which were verified with the trial investigator. MAIN RESULTS: Two controlled trials were identified although data were only available from one trial. The second trial compared two forms of systematic desensitization and reported no discernible differences between them. REVIEWER'S CONCLUSIONS: In spite of encouraging results reported from uncontrolled series of patients there is very limited evidence from controlled trials concerning the effectiveness treatments for vaginismus. Further trials are need to compare therapies with waiting list control and with other therapies. PMID- 11406007 TI - Hormonally impregnated intrauterine systems (IUSs), versus other forms of reversible contraceptives as effective methods of preventing pregnancy. AB - OBJECTIVES: To assess the contraceptive efficacy, tolerability and acceptability of hormonally impregnated intrauterine systems (IUSs) in comparison to other reversible contraceptive methods. SEARCH STRATEGY: Literature was identified through database searches, reference lists and individuals/organisations working in the field. SELECTION CRITERIA: All randomised controlled trials comparing IUSs with other forms of reversible contraceptives and reporting on pre-determined outcomes in women of reproductive years. The primary outcomes were pregnancy due to method/user failure and continuation rate. DATA COLLECTION AND ANALYSIS: The quality assessment of studies and data extraction were completed independently by two blinded reviewers. A quality checklist was designed to identify general methodological and contraceptive specific factors which could bias results. Events per women months and single decrement life table rates were extracted where possible for pregnancy, continuation, adverse events and reasons for discontinuation. Events per total number of women at follow up were collected for hormonal side effects and menstrual disturbance. When appropriate, data were pooled at the same points of follow up to calculate rate ratios in order to determine the relative effectiveness of one method compared to another. For the single decrement life table rates, the rate differences were pooled to determine the absolute difference in effectiveness of one method compared to another. Interventions were only combined if the contraceptive methods were similar. Non hormonal IUDs were divided into three categories for the purpose of comparison with IUSs: IUDs >250mm2 (i.e. CuT 380A IUD and CuT 380 Ag IUD), IUDs <=250mm2 (i.e. Nova-T, Multiload, CuT 200 and CuT 220 IUDs) and non-medicated IUDs. MAIN RESULTS: Nineteen RCTs comparing hormonally impregnated IUSs to a reversible contraceptive method met the inclusion criteria and it was possible to include eight of these in the meta-analyses, four comparing LNG-20 IUSs with non-hormonal IUDs, one comparing the LNG-20 IUS with Norplant-2 and three comparing Progestasert with non-hormonal IUDs. No significant difference was observed between the pregnancy rates for the LNG-20 users and those for the IUD >250mm2 users. However, women using the LNG-20 IUS were significantly less likely to become pregnant than those using the IUD <=250mm2. Women using the LNG-20 IUS were more likely to experience amenorrhoea and device expulsion than women using IUDs >250mm2. LNG-20 users were significantly more likely than all the IUD users to discontinue because of hormonal side effects and menstrual disturbance, which on further breakdown of the data was due to amenorrhoea. When the LNG-20 IUS was compared to Norplant-2, the LNG-20 users were significantly more likely to experience amenorrhoea and oligomenorrhoea, but significantly less likely to experience prolonged bleeding and spotting. No other significant differences were observed. Progestasert users were significantly less likely to become pregnant and less likely to continue on the method than non-medicated IUD users after one year, but no significant differences was noted for these two outcomes when Progestasert users were compared to IUD<=250mm2 users. The only other significant differences found in the meta-analyses were that Progestasert users were less likely to expel the device and more likely to discontinue the method because of menstrual bleeding and pain than users of IUDs <=250mm2. REVIEWER'S CONCLUSIONS: Current evidence suggests LNG-20 IUS users are no more or less likely to have unwanted pregnancies than IUD >250mm2 and Norplant-2 users. The LNG-20 IUS was more effective in preventing either intrauterine or extrauterine pregnancies than IUDs <=250mm2. The contraceptive effectiveness of Progestasert was significantly better than non-medicated IUDs, but no difference was observed when compared to IUDs<=250mm2. Continuation of LNG-20 IUS use was similar to continuation of the non-hormonal IUDs and Norplant-2. Amenorrhoea was the main reason for the discontinuation for the LNG-20 IUS and women should be informed of this prior to starting this method. PMID- 11406008 TI - Local treatments for cutaneous warts. AB - BACKGROUND: Viral warts caused by the human papilloma virus represent one of the most common diseases of the skin. Any area of skin can be affected although the hands and feet are by far the commonest sites. A very wide range of local treatments are available. OBJECTIVES: To assess the effects of different local treatments for cutaneous, non-genital warts in healthy people. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (March 1999), the Skin Group trials register (March 1999), MEDLINE (1966 to August 2000), EMBASE (1980 to August 2000) and a number of other key biomedical and health economics databases. In addition the cited references of all trials identified and key review articles were searched. Pharmaceutical companies involved in local treatments for warts and experts in the field were contacted. SELECTION CRITERIA: Randomized controlled trials of local treatments for cutaneous non-genital viral warts in immunocompetent human hosts were included. DATA COLLECTION AND ANALYSIS: Study selection and assessment of methodological quality were carried out by two independent reviewers. MAIN RESULTS: Forty-nine trials were identified which fulfilled the criteria for inclusion in the review. The evidence provided by these studies was generally weak because of poor methodology and reporting. In 17 trials with placebo groups that used participants as the unit of analysis the average cure rate of placebo preparations was 30% (range 0 to 73%) after an average period of 10 weeks (range 4 to 24 weeks). The best available evidence was for simple topical treatments containing salicylic acid, which are clearly better than placebo. Data pooled from six placebo-controlled trials show a cure rate of 144/191 (75%) compared with 89/185 (48%) in controls, odds ratio 3.91 (95% confidence interval 2.40 to 6.36), random effects model. Most of the bigger trials of cryotherapy studied different regimens rather than comparing cryotherapy with other treatments or placebo. Pooled data from two small trials that included cryotherapy and placebo or no treatment, showed no significant difference in cure rates. In two other trials comparing cryotherapy with salicylic acid no significant difference in efficacy was demonstrated. There was no consistent evidence for the effectiveness of intralesional bleomycin. Four studies, using warts rather than individuals as the unit of analysis, had widely varying results which could not be meaningfully pooled. There was some evidence for the efficacy of dinitrochlorobenzene, a potent contact sensitizer. Pooled data from two small studies comparing dinitrochlorobenzene with placebo showed cure rates of 32/40 (80%) and 17/40 (43%) respectively, odds ratio 5.42 (95% confidence interval 1.99 to 14.75), random effects model. Only limited evidence was found for the efficacy of topical 5-fluorouracil, intralesional interferons and photodynamic therapy. Bleomycin, dinitrochlorobenzene, 5-fluorouracil, interferons and photodynamic therapy are potentially hazardous or toxic treatments. REVIEWER'S CONCLUSIONS: There is a considerable lack of evidence on which to base the rational use of the local treatments for common warts. The reviewed trials are highly variable in method and quality. Cure rates with placebo preparations are variable but nevertheless considerable. There is certainly evidence that simple topical treatments containing salicylic acid have a therapeutic effect. There is less evidence for the efficacy of cryotherapy and no convincing evidence that it is any more effective than simple topical treatments. Dinitrochlorobenzene appears to be effective but no more so than the safer, simpler and cheaper topical treatments containing salicylic acid. The benefits and risks of 5-fluorouracil, bleomycin, interferons and photodynamic therapy remain to be determined. PMID- 11406009 TI - Plasma exchange for Guillain-Barre syndrome. AB - BACKGROUND: Guillain-Barre syndrome is an acute symmetric usually ascending and usually paralysing illness due to inflammation of peripheral nerves. It is thought to be caused by autoimmune factors, such as antibodies. Plasma exchange removes antibodies and other potentially injurious factors from the blood stream. It involves connecting the patient's blood circulation to a machine which exchanges the plasma for a substitute solution, usually albumin. Several studies have evaluated plasma exchange for Guillain-Barre syndrome. OBJECTIVES: To systematically review the evidence concerning the efficacy of plasma exchange for treating Guillain-Barre syndrome. SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Trial Register for randomised trials concerning plasma exchange in Guillain-Barre syndrome, search of the bibliographies of identified papers and enquiry from the authors of the papers. SELECTION CRITERIA: Randomised and quasi-randomised trials of plasma exchange versus sham exchange or supportive treatment. DATA COLLECTION AND ANALYSIS: Potentially relevant papers were scrutinised by two reviewers and the selection of eligible studies was agreed by them and a third reviewer. Data were extracted by one reviewer and checked by a second reviewer. Some missing data were obtained from the authors of studies. MAIN RESULTS: Six eligible trials concerning 649 patients were identified, all comparing plasma exchange versus supportive treatment alone. Primary outcome measures ~Bullet~Time to recover walking with aid In the only two trials for which this measure was reported the median time to recover this ability was faster in the plasma exchange than the control group. ~Bullet~Time to onset of motor recovery in mildly affected patients In the one trial for which this measure was available the time was significantly shortened in the plasma exchange group. Secondary outcome measures ~Bullet~Improvement in disability grade at 4 weeks In five trials, there were significantly more patients who had improved by one disability grade or more in the plasma exchange group as compared to the control group. Patients treated with plasma exchange fared significantly better in the following secondary outcome measures: time to recover walking without aid, percentage of patients requiring artificial ventilation, duration of ventilation, full muscle strength recovery after one year, and severe sequelae after one year. There were less patients with infectious events and cardiac arrhythmias in the plasma exchange than the control group. Subgroup analyses Plasma exchange was beneficial in patients with mild, moderate and severe (needing ventilation) Guillain-Barre syndrome. It was beneficial in patients with a disease duration of seven or less days and also in those with disease lasting more than seven days. However, in the only trial that enrolled patients up to 30 days from disease onset, the benefit of plasma exchange in patients treated after seven days was less apparent. Type of treatment Single studies showed that two plasma exchanges were significantly superior to none for mild Guillain-Barre syndrome and four to two for moderate Guillain-Barre syndrome but that six were not superior to four for severe Guillain-Barre syndrome requiring ventilation. One study suggested that continuous flow plasma exchange was significantly superior to intermittent flow. Another study found no significant difference between the two techniques. The same study found a significantly higher rate of adverse events with fresh frozen plasma as the replacement fluid than albumin. REVIEWER'S CONCLUSIONS: Plasma exchange is the first and only treatment that has been proven to be superior to supportive treatment alone in Guillain-Barre syndrome. Consequently, plasma exchange should be regarded as the treatment against which new treatments, such as intravenous immunoglobulin, should be judged. In mild Guillain-Barre syndrome two sessions of plasma exchange are superior to none. In moderate Guillain-Barre syndrome four sessions are superior to two. In severe Guillain Barre syndrome six sessions are no better than four. Continuous flow plasma exchange machines may be superior to intermittent flow machines and albumin to fresh frozen plasma as the exchange fluid. Plasma exchange is more beneficial when started within seven days after disease onset rather than later, but was still beneficial in patients treated up to 30 days after disease onset. The value of plasma exchange in children less than 12 years old is not known. PMID- 11406010 TI - Erythromycin for feeding intolerance in preterm infants. AB - BACKGROUND: Functional immaturity of gastrointestinal motility predisposes preterm infants to feeding intolerance. Motilin, a gastrointestinal peptide, stimulates propagative contractile activity during phase III of the migratory motor complex in the interdigestive state. Erythromycin (EM) is a motilin agonist with prokinetic effect at low doses (1-3mg/kg). OBJECTIVES: To evaluate the effectiveness of EM in promoting gastrointestinal motility in preterm infants with feeding intolerance and assess clinically significant adverse effects associated with its use. SEARCH STRATEGY: Systematic literature search in accordance with the Cochrane Neonatal Collaborative Review Group search strategy. Randomized and quasi-randomized controlled trials of EM use, at any dose, in preterm infants to promote gastrointestinal motility were identified by searching MEDLINE, EMBASE, CINAHL, the Cochrane Library, reference lists of published studies, personal files, and abstracts published in Pediatric Research. SELECTION CRITERIA: Randomized controlled trials of oral or intravenous EM use at dose range of 3 to 12 mg/kg/day in preterm infants less than or equal to 36 weeks gestational age with feeding tolerance were included in this review. DATA COLLECTION AND ANALYSIS: Data regarding the primary clinical outcome of days to achieve full enteral feeding were compared among studies. Data on secondary outcomes including adverse effects associated with the use of EM (diarrhea, nosocomial infections, cardiac arrhythmias, or theophylline toxicity), duration of parenteral nutrition, weight gain, incidence of necrotizing enterocolitis (NEC), hypertrophic pyloric stenosis, and length of hospital stay were assessed. MAIN RESULTS: Two randomized controlled studies of EM use in preterm infants for improving gastrointestinal motility were identified. Since both studies involved preterm infants treated with EM at dose >12mg/kg/day at commencement of feeding, they did not meet inclusion criteria defined a priori for this review. There was no statistically significant difference in the incidence of NEC (RR 0.59, 95%CI 0.11, 3.01; RD -0.021, 95%CI -0.087, 0.045). No statistically significant difference was noted in days to achieve full enteral feeds, length of hospital stay, and adverse events between groups. REVIEWER'S CONCLUSIONS: EM at antimicrobial doses may not be effective in preterm infants with feeding intolerance. Further studies are needed to determine whether EM in lower doses is effective as a prokinetic agent in such infants. PMID- 11406011 TI - Orthotic devices for tennis elbow. AB - BACKGROUND: Lateral epicondylitis (tennis elbow) is a frequently reported condition. A wide variety of treatment strategies has been described. As of yet, no optimal strategy has been identified. OBJECTIVES: The objective of this review was to assess the effectiveness of orthotic devices for treatment of tennis elbow. SEARCH STRATEGY: An electronic database search was conducted using Medline, Embase, Cinahl, the Cochrane Controlled Trial Register, Current Contents and reference lists from all retrieved articles. Experts on the subjects were approached for additional trials. SELECTION CRITERIA: All randomised clinical trials (RCT) describing individuals with diagnosed lateral epicondylitis and comparing the use of an orthotic device as a treatment strategy were evaluated for inclusion. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the validity of the included trials and extracted data on relevant outcome measures. Dichotomous outcomes were expressed as Relative Risks (RRs) and continuous outcomes as Standardised Mean Differences (SMD), both with corresponding 95% confidence intervals (95% CI). Statistical pooling and subgroup analyses were intended MAIN RESULTS: Five small-size RCTs (N per group 7-49) were included. Validity score ranged from 3-9 positive items out of 11. Subgroup analyses were not performed due to the small number of trials. The limited number of included trials present few outcome measures and limited long-term results. Pooling was not possible due to large heterogeneity amongst trials. REVIEWER'S CONCLUSIONS: No definitive conclusions can be drawn concerning effectiveness of orthotic devices for lateral epicondylitis. More well-designed and well-conducted RCTs of sufficient power are warranted. PMID- 11406012 TI - Interventions for smoking cessation in hospitalised patients. AB - BACKGROUND: An admission to hospital provides an opportunity to help people stop smoking. Individuals may be more open to help at a time of perceived vulnerability, and may find it easier to quit in an environment where smoking is restricted or prohibited. Providing smoking cessation services during hospitalisation may help more people to attempt and sustain a quit attempt. OBJECTIVES: To determine the effectiveness of interventions for smoking cessation in hospitalised patients. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group register, CINAHL and the Smoking and Health database for studies of interventions for smoking cessation in hospitalised patients. SELECTION CRITERIA: Randomised and quasi-randomised trials of behavioural, pharmacological or multicomponent interventions to help patients stop smoking conducted with hospitalised patients who were current smokers or recent quitters. We excluded studies of patients admitted for psychiatric disorders or substance abuse, those that did not report abstinence rates and those with follow-up of less than six months. DATA COLLECTION AND ANALYSIS: Two authors extracted data independently for each paper, with disagreements resolved by consensus. MAIN RESULTS: Intensive intervention (inpatient contact plus follow-up for at least one month) was associated with a significantly higher quit rate compared to control (Peto Odds Ratio 1.82, 95% CI 1.49-2.22). There was insufficient evidence to judge the effect of interventions delivered only during the hospital stay. Although the interventions increased quit rates irrespective of whether nicotine replacement therapy was used, the results for nicotine replacement therapy (NRT) were compatible with other data indicating that it increases quit rates. There was no strong evidence that clinical diagnosis affected the likelihood of quitting. REVIEWER'S CONCLUSIONS: High intensity behavioural interventions that include at least one month of follow-up contact are effective in promoting smoking cessation in hospitalised patients. The findings of the review were compatible with research in other settings showing that NRT increases quit rates. PMID- 11406013 TI - Conservative management for post prostatectomy urinary incontinence. AB - BACKGROUND: Urinary incontinence after prostatectomy is a common problem. Conservative management of this condition includes pelvic floor muscle training, biofeedback, electrical stimulation using a rectal electrode, transcutaneous electrical nerve stimulation, or a combination of methods. OBJECTIVES: To assess the effects of conservative management for urinary incontinence after transurethral, suprapubic, radical retropubic or perineal prostatectomy. SEARCH STRATEGY: The Cochrane Incontinence Group's trials register, Medline, Cinahl, Embase, PsycLit and ERIC all up to January 1999, and reference lists of relevant articles. We contacted investigators to locate studies and we handsearched the following conference proceedings: American Urological Association (1989-1999); Society of Urologic Nurses and Associates (1991-1998); Wound Ostomy and Continence Nurses (1996-1999); and International Continence Society (1980-1998). Date of most recent searches: January 1999. SELECTION CRITERIA: Randomised or quasi-randomised trials which evaluated conservative management aimed at improving urinary continence after prostatectomy. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the methodological quality of studies and abstracted data from included trials onto a standard form. MAIN RESULTS: Only five randomised trials were identified which included 365 men, each evaluating different treatments, and all studying men after radical prostatectomy. The trials were of moderate quality and data were not available for many of the pre stated outcomes. Confidence intervals for both dichotomous and continuous data were wide; it was not possible to reliably identify or rule out a useful effect. Men's symptoms tended to improve over time, irrespective of management. REVIEWER'S CONCLUSIONS: The value of the various approaches to conservative management of post prostatectomy incontinence remains uncertain. Further well designed trials are needed. PMID- 11406014 TI - Aciclovir for Bell's palsy (idiopathic facial paralysis). AB - BACKGROUND: The most common disorder of the facial nerve is acute idiopathic facial paralysis or Bell's palsy and there may be significant morbidity or incomplete recovery associated with severe cases. Although the cause remains unknown, recent evidence suggests a possible association with Herpes simplex virus (HSV) infection. To test this hypothesis clinically four clinical trials have used aciclovir, an antiviral agent, either alone or in combination with corticosteroids to treat Bell's palsy. OBJECTIVES: To assess the efficacy of aciclovir for treating Bell's palsy and to evaluate any adverse effects of the drug treatment. SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Group register, MEDLINE, EMBASE and LILACS databases for randomised trials. We also contacted authors of identified trials. SELECTION CRITERIA: Randomised or quasi-randomised trials of aciclovir therapy, alone or in combination with any other drug, in patients with Bell's palsy. DATA COLLECTION AND ANALYSIS: We identified four randomised trials. One author extracted the data and the other checked them. We wrote to all the authors of the trials identified. No additional data were obtained. MAIN RESULTS: Only two studies met our inclusion criteria and provided results from 200 patients. One study evaluated aciclovir with corticosteroid versus steroid alone and the other study evaluated aciclovir alone versus corticosteroid. PRIMARY OUTCOME Proportion of patients with incomplete recovery after one year: These data were not available. However an analysis was performed on data reported at the end of the study period, three (De Diego 1998) or four months (Adour 1996) after the start of treatment. The results from the Adour study significantly favoured the treatment group whilst the De Diego study significantly favoured the control group. Adverse events: This analysis was not performed as the data were not reported. Complete facial paralysis six months after start of treatment: Only one patient had complete paralysis upon entering only one of the studies (De Diego 1998). This patient was assigned to the control group and the level of recovery attained was not reported. Motor synkinesis/Crocodile tears one year after start of treatment: Data were available up to a maximum of four months after onset of paralysis. One study reported a significant difference between the treatment groups in favour of the aciclovir group and the other demonstrated an inconclusive result. REVIEWER'S CONCLUSIONS: More data are needed from a large multicentre randomised controlled and blinded study with at least 12 months' follow up before a definitive recommendation can be made regarding the routine use of aciclovir in Bell's palsy. PMID- 11406015 TI - Interventions for preventing obesity in children. AB - BACKGROUND: The prevalence of obesity and overweight is increasing world-wide. Obesity in children impacts on their health in both the short and longer term. Obesity prevention strategies are poorly understood. OBJECTIVES: To assess the effectiveness of interventions designed to prevent obesity in childhood. SEARCH STRATEGY: Electronic databases were searched from 1985 to October 1999. SELECTION CRITERIA: Data from RCTs and non-randomised trials with concurrent control group were included. studies with follow up of one year minimum were selected, A priori, this was subsequently amended to include studies with minimum follow up of three months. The possible susceptibility of post hoc questions to bias is addressed. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed study quality. MAIN RESULTS: Seven studies were included, three long-term and four short- term. The studies included were diverse in terms of study design and quality, target population, theoretical underpinning of intervention approach, and outcome measures. As such, it was not possible to combine study findings using statistical methods. REVIEWER'S CONCLUSIONS: The findings of the review suggest that currently there is limited quality data on the effectiveness of obesity prevention programs and as such no generalisable conclusions can be drawn. The need for well-designed studies which examine a range of interventions remains a priority. PMID- 11406016 TI - Desmopressin for minimising perioperative allogeneic blood transfusion. AB - BACKGROUND: Public concerns regarding the safety of transfused blood have prompted re-consideration of the use of allogeneic (from an unrelated donor) red blood cell (RBC) transfusion, and a range of techniques designed to minimise transfusion requirements. OBJECTIVES: To examine the evidence for the efficacy of desmopressin (1-deamino-8-D-arginine-vasopressin), in reducing perioperative blood loss and the need for red cell transfusion in patients who do not have congenital bleeding disorders. SEARCH STRATEGY: Articles were identified by: computer searches of OVID MEDLINE, EMBASE, and Current Contents (to August 2000) and web sites of international health technology assessment agencies (to May 1998). References in the identified trials and review articles were checked and authors contacted to identify additional studies. SELECTION CRITERIA: Randomised controlled parallel group trials in which adult patients, scheduled for non urgent surgery, were randomised to DDAVP, or to a control group, who did not receive the intervention. DATA COLLECTION AND ANALYSIS: Trial quality was assessed using criteria proposed by Schulz et al. (1995) and Jadad et al. (1996). The principal outcomes were: the number of patients exposed to red cells, and the amount of blood transfused. Other clinical outcomes are detailed in the review. MAIN RESULTS: Fourteen trials of DDAVP (N=1034) reported data on the proportion of patients exposed to allogeneic RBC transfusion. In subjects treated with DDAVP the relative risk of exposure to peri-operative allogeneic blood transfusion was 0.98 (95%CI: 0.88 to 1.10) compared with control. In DDAVP-treated patients the relative risk of requiring re-operation due to bleeding was 0.56 (95%CI: 0.18 to 1.73). There was no statistically significant effect overall for mortality and non-fatal myocardial infarction in DDAVP-treated patients compared with control (RR=1.53: 95%CI: 0.58 to 4.05) and (RR=1.52: 95%CI: 0.67 to 3.49) respectively. REVIEWER'S CONCLUSIONS: There is no convincing evidence that desmopressin minimises perioperative allogeneic RBC transfusion in patients who do not have congenital bleeding disorders. These data suggest that there is no benefit of using DDAVP as a means of minimising perioperative allogeneic RBC transfusion. This meta-analysis had 90% power to detect a relative risk reduction of at least 17% for receiving a red cell transfusion at alpha = 0.05 (two-sided). PMID- 11406017 TI - Low protein diets for chronic renal failure in non diabetic adults. AB - BACKGROUND: For more than fifty years, low protein diets have been proposed to patients with kidney failure. However, the effects of these diets in preventing severe renal failure and the need for maintenance dialysis have not been resolved. OBJECTIVES: To determine the efficacy of low protein diets in delaying the need to start maintenance dialysis. SEARCH STRATEGY: Medline and Embase search from January 1966 through to June 1999. Congress abstracts (American Society of Nephrology since 1990, European Dialysis Transplant Association since 1985, International Society of Nephrology since 1987). Direct contacts with investigators. SELECTION CRITERIA: Randomised trials comparing two different levels of protein intake in adult patients suffering from moderate to severe renal failure, followed for at least one year. Diabetic nephropathy patients were excluded. DATA COLLECTION AND ANALYSIS: Seven trials were selected from over 40 studies since 1975. A total of 1494 patients were analysed, 753 had received reduced protein intake and 741 a higher protein intake. Collection of the number of "renal deaths" defined as the need for starting dialysis, the death of a patient or a kidney transplant during the trial. MAIN RESULTS: Two hundred and forty two renal deaths were recorded, 101 in the low protein diet and 141 in the higher protein diet group, giving an odds ratio of 0.62 with a 95% confidence interval of 0.46 to 0.83 (p=0.006). To avoid one renal death, four to 56 patients need to be treated with a low protein diet during one year. REVIEWER'S CONCLUSIONS: Reducing protein intake in patients with chronic renal failure reduces the occurrence of renal death by about 40% as compared with higher or unrestricted protein intake. The optimal level of protein intake cannot be confirmed from these studies. PMID- 11406018 TI - Dietary treatment for familial hypercholesterolaemia. AB - BACKGROUND: Familial hypercholesterolaemia is an inherited disorder characterised by a raised blood cholesterol, the presence of xanthomatosis and premature ischaemic heart disease. The aim of treatment is the reduction of blood LDL cholesterol concentrations in order to reduce the risk of ischaemic heart disease. Current treatment is based on a cholesterol lowering diet alone or in combination with drug therapy. Many of the drugs found to be effective in treating adults with this disease are not licensed for use in children, therefore diet is the main treatment of children with familial hypercholesterolaemia. In addition to the cholesterol-lowering diet, several other dietary interventions have been suggested and consensus has yet to be reached on the most appropriate dietary treatment for children and adults with familial hypercholesterolaemia. OBJECTIVES: To examine the evidence that in children and adults with familial hypercholesterolaemia, a cholesterol lowering diet is more effective at lowering cholesterol and reducing incidence of ischaemic heart disease than no intervention or than other dietary interventions. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Trials Register, a specialist trials register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Additional studies were identified from handsearching the Journal of Inherited Metabolic Disease (from inception, 1978 to 2000) and from the reference lists of identified studies. SELECTION CRITERIA: Randomised controlled trials (RCTs), both published and unpublished, where a cholesterol lowering diet in children and adults with familial hypercholesterolaemia has been compared to other forms of dietary treatment or to no dietary intervention. Trials which include patients with familial hypercholesterolaemia alongside patients with non-familial hypercholesterolaemia were only included if the group of familial patients was well defined and the results for these patients were available. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the trial eligibility and methodological quality and one reviewer extracted the data, with independent verification of data extraction by a colleague. MAIN RESULTS: Only short term outcomes could be assessed in this review due to the length of the five eligible studies. Compliance to treatment, quality of life, mortality and evidence of ischaemic or atheromatous disease were not assessed in the studies identified. No differences were found between the cholesterol-lowering diet and all other diets for all of the short term outcomes assessed. REVIEWER'S CONCLUSIONS: No conclusions can be made about the effectiveness of the cholesterol-lowering diet, or any of the other dietary interventions suggested for familial hypercholesterolaemia, due to the lack of adequate data. A large, parallel, randomised controlled trial is needed to investigate the effectiveness of the cholesterol-lowering diet and other dietary interventions for FH. It is also possible that data from trials including subjects with both familial and non-familial hypercholesterolaemia could alter the results of future updates of this review and until further evidence is available current dietary treatment of FH should continue to be observed and monitored with care. PMID- 11406019 TI - Surgical versus non-surgical interventions for vocal cord nodules. AB - BACKGROUND: Vocal cord nodules are bilateral swellings of the mid-portion of the membranous vocal folds. They are of variable size and are characterised histologically by thickening of the epithelium with a variable degree of inflammatory action in the underlying superficial lamina propria. They characteristically produce hoarseness. Treatment of vocal cord nodules aims to eliminate or reduce this hoarseness. OBJECTIVES: To assess the effectiveness of surgery versus non-surgical interventions for vocal cord nodules. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register (CCTR) and Medline (1966-2000), Embase (1974-2000), Biological Abstracts (1970-2000), Biological Abstracts RRM (Reports, Reviews and Meetings) on CD-ROM (1989-2000) and review articles. SELECTION CRITERIA: Randomised and quasi-randomised trials comparing any surgical intervention for vocal cord nodules with non-surgical treatment or no treatment. DATA COLLECTION AND ANALYSIS: No suitable trials were identified. MAIN RESULTS: No studies fulfilled the inclusion criteria. REVIEWER'S CONCLUSIONS: There is a need for high quality randomised controlled trials to evaluate the effectiveness of surgical and non-surgical treatment of vocal cord nodules. PMID- 11406020 TI - Elective delivery in diabetic pregnant women. AB - BACKGROUND: In pregnancies complicated by diabetes the major concerns during the third trimester are fetal distress and the potential for birth trauma associated with fetal macrosomia. OBJECTIVES: The objective of this review was to assess the effect of a policy of elective delivery, as compared to expectant management, in term diabetic pregnant women, on maternal and perinatal mortality and morbidity. SEARCH STRATEGY: We searched the Cochrane Pregnancy and Childbirth Group trials register and the Cochrane Controlled Trials Register (last searched February 2001). SELECTION CRITERIA: All available randomized controlled trials of elective delivery, either by induction of labour or by elective caesarean section, compared to expectant management in diabetic pregnant women at term. DATA COLLECTION AND ANALYSIS: The reports of the only available trial were analysed independently by the three co-reviewers to retrieve data on maternal and perinatal outcomes. Results are expressed as relative risks (RR) and 95% confidence intervals (CI). MAIN RESULTS: The participants in the one trial included in this review were 200 insulin-requiring diabetic women. Most had gestational diabetes, except 13 women with type 2 pre-existing diabetes (class B). The trial compared a policy of active induction of labour at 38 completed weeks of pregnancy, to expectant management until 42 weeks. The risk of caesarean section was not statistically different between groups (relative risk (RR) 0.81, 95% confidence interval (CI) 0.52 - 1.26). The risk of macrosomia was reduced in the active induction group (RR 0.56, 95% CI 0.32 - 0.98) and three cases of mild shoulder dystocia were reported in the expectant management group. No other perinatal morbidity was reported. REVIEWER'S CONCLUSIONS: There is very little evidence to support either elective delivery or expectant management at term in pregnant women with insulin-requiring diabetes. Limited data from a single randomized controlled trial suggest that induction of labour in women with gestational diabetes treated with insulin reduces the risk of macrosomia. Although the small sample size does not permit one to draw conclusions, the risk of maternal or neonatal morbidity was not modified. Women's views on elective delivery and on prolonged surveillance and treatment with insulin should be assessed in future trials. PMID- 11406021 TI - Dietary advice for illness-related malnutrition in adults. AB - BACKGROUND: Illness-related malnutrition has been reported in 10-55% of people in hospital and in ill people in the community in areas of food sufficiency. It has been suggested that dietary counseling to encourage the use of energy- and protein-rich foods should be used in preference to oral nutritional supplements in the management of illness-related malnutrition. OBJECTIVES: To examine the evidence that dietary advice, to improve nutritional intake, in adults with illness-related malnutrition can improve survival, weight and anthropometry and to estimate the size of any additional effect of whole protein nutritional supplements when given in combination with dietary advice. SEARCH STRATEGY: Six electronic databases were searched (Cochrane Library, Medline, EMBASE, CINAHL, ERIC and Dissertation Abstracts). Additional studies were sought by contacting dietitians, clinicians and the manufacturers of nutritional supplements. SELECTION CRITERIA: All randomised controlled trials of dietary advice compared with no advice, oral nutritional supplements and dietary advice plus oral nutritional supplements in people with illness-related malnutrition. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed the trial eligibility, methodological quality and extracted the data. MAIN RESULTS: Fifteen trials (16 comparisons) met the inclusion criteria. An additional group of six trials were identified during searching which compared dietary advice plus supplements if required with no advice. These trials have been included in the review as an additional comparison. The review includes 1185 randomised participants from a variety of clinical backgrounds. The duration of follow-up was from six weeks to 12 months. Nine trials reported clearly concealed randomisation and allocation and two trials reported blinding of outcome assessment. Nine trials reported outcomes in an unusable format. Gain in weight was significantly higher for groups consuming supplements with or without advice, compared with advice alone. Energy intake was significantly improved in those consuming supplements compared with those receiving advice alone. There was insufficient data on group differences in functional outcomes and mortality. REVIEWER'S CONCLUSIONS: This review highlights the lack of evidence for the provision of dietary advice in the management of illness-related malnutrition. The available data suggest that oral nutritional supplements have a greater role than dietary advice in the improvement of body weight and energy intake. A large adequately powered randomised controlled trial is needed to compare the efficacy of different forms of therapy to increase dietary intake in people with illness related malnutrition and to examine the impact of this on clinical function and survival. PMID- 11406022 TI - Pulse oximetry for perioperative monitoring. AB - BACKGROUND: Monitoring with pulse oximetry might improve patient outcome by enabling an early diagnosis and consequently, correction of perioperative events that might cause postoperative complications or even death. Only a few randomised clinical trials of pulse oximetry have been performed during anaesthesia and in the recovery room which describe perioperative hypoxaemic events, postoperative cardiopulmonary complications and cognitive dysfunction. OBJECTIVES: To study the effect of perioperative monitoring with pulse oximetry to clearly identify the adverse outcomes that might be prevented or improved by the use of pulse oximetry. SEARCH STRATEGY: Trials were identified by computerised searches of the Cochrane Library, MEDLINE, EMBASE, and by checking the reference lists of trials and review articles. SELECTION CRITERIA: All controlled trials that randomised patients to either pulse oximetry or no pulse oximetry during the perioperative period, including the operating and recovery room. DATA COLLECTION AND ANALYSIS: We collected data in relation to events detectable by pulse oximetry, any serious complications that occurred during anaesthesia or in the postoperative period, intra- or postoperative mortality, and duration of recovery or intensive care stay. Formal statistical synthesis of individual trials was not performed in view of the variety of outcomes studied. MAIN RESULTS: Searching identified six reports; four studies with data from a total of 21,773 patients were considered eligible for analysis. Only two studies specifically addressed the outcomes in question; both found no effect on the rate of postoperative complications using perioperative pulse oximetry. Two studies used hypoxaemia detectable by pulse oximetry to assess the value of perioperative monitoring, although outcomes were not given. It was found that hypoxaemia was reduced in the pulse oximetry group both in the operating theatre and in the recovery room. During observation in the recovery room, the incidence of hypoxaemia in the pulse oximetry group was 1.5-3 times less. The postoperative cognitive function using the Wechsler memory scale and continuous reaction time was independent of perioperative monitoring with pulse oximetry. The other study showed that postoperative complications occurred in 10% of the patients in the oximetry group and in 9.4% in the control group. The two groups did not differ in cardiovascular, respiratory, neurologic, or infectious complications. The duration of hospital stay was a median of 5 days in both groups, and an equal number of in-hospital deaths was registered in the two groups. REVIEWER'S CONCLUSIONS: The studies confirmed that pulse oximetry can detect hypoxaemia and related events. However, we have found no evidence that pulse oximetry affects the outcome of anaesthesia. The conflicting subjective and objective results of the studies, despite an intense, methodical collection of data from a relatively large population, indicates that the value of perioperative monitoring with pulse oximetry is questionable in relation to improved reliable outcomes, effectiveness and efficiency. PMID- 11406023 TI - Antidepressant drug treatment for postnatal depression. AB - BACKGROUND: Postnatal depression is a common disorder, which can have profound short and long term effects on maternal morbidity, the new infant and the family as a whole. Social factors appear to be particularly important in the aetiology and prognosis of postnatal depression and treatment is often largely social support and psychological interventions. It is not known whether antidepressants are an effective and safe choice for treatment of this disorder. OBJECTIVES: To evaluate the effectiveness of different antidepressant drugs and compare their effectiveness with other forms of treatment. To assess any adverse effects of antidepressants in the mother or the nursing baby. SEARCH STRATEGY: The registers of clinical trials maintained and updated by the Cochrane Depression, Anxiety and Neurosis Group and the Cochrane Pregnancy and Childbirth Group were searched. Other databases (outlined below) were also searched and contacts were made with pharmaceutical companies and experts in the field. SELECTION CRITERIA: All trials were considered in which women with depression in the first six months postpartum were randomised to receive antidepressants alone or in combination with another treatment, or to receive any other treatment including placebo. DATA COLLECTION AND ANALYSIS: Data was extracted independently from the trial reports by the reviewers. Missing information was requested from investigators wherever possible. Data was sought to allow an "intention to treat" analysis. MAIN RESULTS: Only one trial could be included in this review, leaving most of the objectives of the review unfulfilled. Appleby et al (1997) reported that Fluoxetine was significantly more effective than placebo and, after an initial session of counselling, as effective as a full course of cognitive-behavioural counselling in the treatment of postnatal depression. There was no interaction between medication and counselling. REVIEWER'S CONCLUSIONS: Women with postnatal depression can be effectively treated with fluoxetine, which is as effective as a course of cognitive-behavioural counselling in the short-term. However, more trials with a longer follow-up period are needed to compare different antidepressants in the treatment of postnatal depression, and to compare antidepressant treatment with psychosocial interventions. This is an area that has been neglected despite the large public health impact described above. PMID- 11406024 TI - Parent-training programmes for improving maternal psychosocial health. AB - BACKGROUND: Mental health problems are common, and there is evidence from a range of studies to suggest that a number of factors relating to maternal psychosocial health can have a significant effect on the mother-infant relationship, and that this can have consequences for both the short and long-term psychological health of the child. The use of parenting programmes is increasing in the UK and evidence of their effectiveness in improving outcomes for mothers is now required. OBJECTIVES: The objective of this review is to address whether group based parenting programmes are effective in improving maternal psychosocial health including anxiety, depression, and self-esteem. SEARCH STRATEGY: A range of biomedical, social science, educational and general reference electronic databases were searched including MEDLINE, EMBASE CINAHL, PsychLIT, ERIC, ASSIA, Sociofile and the Social Science Citation Index. Other sources of information included the Cochrane Library (SPECTR, CENTRAL), and the National Research Register (NRR). SELECTION CRITERIA: Only randomised controlled trials were included in which participants had been randomly allocated to an experimental and a control group, the latter being either a waiting-list, no-treatment or a placebo control group. Studies had to include at least one group-based parenting programme, and one standardised instrument measuring maternal psychosocial health. DATA COLLECTION AND ANALYSIS: A systematic critical appraisal of all included studies was undertaken using a modified version of the Journal of the American Medical Association (JAMA) published criteria. The treatment effect for each outcome in each study was standardised by dividing the mean difference in post-intervention scores for the intervention and treatment group, by the pooled standard deviation, to produce an effect size. Where appropriate the results were then combined in a meta-analysis using a fixed-effect model, and 95% confidence intervals were used to assess the significance of the findings. MAIN RESULTS: A total of 23 studies were included in the review but only 17 provided sufficient data to calculate effect sizes. The 17 studies provided a total of 59 assessments of outcome on a range of aspects of psychosocial functioning including depression, anxiety, stress, self-esteem, social competence, social support, guilt, mood, automatic thoughts, dyadic adjustment, psychiatric morbidity, irrationality, anger and aggression, mood, attitude, personality, and beliefs. There was only sufficient data, however, on five outcomes (depression; anxiety/stress; self-esteem; social support; and relationship with spouse/marital adjustment) to combine the results in a meta-analysis. The meta-analyses show statistically significant results favouring the intervention group as regards depression; anxiety/stress; self-esteem; and relationship with spouse/marital adjustment. The meta-analysis of the social support data, however, showed no evidence of effectiveness. These results suggest that parenting programmes, irrespective of the type (or content) of programme, can be effective in improving important aspects of maternal psycho-social functioning. Of the data summarising the effectiveness of the different types of parenting programmes, which it was not possible to combine in a meta-analysis, approximately 22% of the outcomes measured, showed significant differences between the intervention group and the control group. A further 40% showed medium to large non-significant differences favouring the intervention group. Approximately one-third of outcomes showed small non-significant differences or no evidence of effectiveness. A meta analysis of the follow-up data on three outcomes was also conducted - depression, self-esteem and relationship with spouse/marital adjustment. The results show that there was a continued improvement in self-esteem, depression and marital adjustment at follow-up, although the latter two findings were not statistically significant. REVIEWER'S CONCLUSIONS: It is suggested that parenting programmes can make a significant contribution to short-term psychosocial health in mothers, and that the limited follow-up data available suggest that these are maintained over time. However, the overall paucity of long-term follow-up data points to the need for further evidence concerning the long-term effectiveness of parenting programmes on maternal mental health. Furthermore, it is suggested that some caution should be exercised before the results are generalised to parents irrespective of the level of pathology present, and that further research is still required. PMID- 11406025 TI - Diaphragm versus diaphragm with spermicides for contraception. AB - BACKGROUND: The diaphragm is usually used with a spermicide. However, some practitioners have suggested that spermicides offer no additional contraceptive protection and have advocated alternative guidelines for the use of diaphragms. OBJECTIVES: The objective of this review was to compare the effectiveness, safety and acceptability of the diaphragm with and without spermicide. SEARCH STRATEGY: We searched MEDLINE, EMBASE, POPLINE, CINAHL, the Cochrane Controlled Trials Register, and reference lists of relevant articles. In addition, we contacted experts in the field to identify unpublished studies. SELECTION CRITERIA: Randomized controlled trials comparing women of reproductive age using the diaphragm with and without spermicide as the sole contraceptive method that reported clinical outcomes. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data on outcomes and trial characteristics and any discrepancies were resolved by consensus or by consultation with the third reviewer. The results of the one identified study are presented descriptively. MAIN RESULTS: We identified only one study. No significant difference was found in the pregnancy rates (with typical use or consistent use) or discontinuation rates between the diaphragm-with-spermicide and diaphragm-without-spermicide groups. There was a trend towards higher pregnancy rates in the diaphragm-without spermicide group. However, this study failed to recruit the planned number of participants and was consequently underpowered. REVIEWER'S CONCLUSIONS: As only one underpowered study was identified, we cannot distinguish between the contraceptive effectiveness of the diaphragm with and without spermicide. We cannot draw any conclusion at this point, further research is needed. PMID- 11406026 TI - Biphasic versus monophasic oral contraceptives for contraception. AB - OBJECTIVES: To compare biphasic oral contraceptives with monophasic oral contraceptives in terms of efficacy, cycle control, and discontinuation due to side effects. Our a priori hypotheses were: (a) biphasic oral contraceptives are less effective in preventing pregnancy than are monophasic oral contraceptives, and (b) biphasic oral contraceptives cause more side effects, give poorer cycle control, and have lower continuation rates. SEARCH STRATEGY: We searched computerized databases MEDLINE, EMBASE, Popline and the Cochrane Controlled Trial Register. Additionally we searched the reference lists of all potentially relevant articles and book chapters. We also contacted the authors of relevant studies and pharmaceutical companies in Europe and the U.S. SELECTION CRITERIA: We included randomized controlled trials comparing any biphasic oral contraceptive with any monophasic oral contraceptive when used to prevent pregnancy. DATA COLLECTION AND ANALYSIS: We examined the studies found during the various literature searches for possible inclusion and assessed their methodological quality using the Cochrane guidelines. We contacted the authors of all included studies and of possibly randomized studies for supplemental information about the study methods and outcomes. We entered the data in RevMan 3.1, imported the data into RevMan 4.1, and calculated Peto odds ratios for the incidence of intermenstrual bleeding, absence of withdrawal bleeding, and study discontinuation due to intermenstrual bleeding. MAIN RESULTS: Only one trial of limited quality compared a biphasic and monophasic preparation. Percival-Smith et al. (1990) examined 533 user cycles of a biphasic pill (norethindrone 500 mcg/ethinyl estradiol 35 mcg for ten days, followed by norethindrone 1000 mcg/ethinyl estradiol 35 mcg for eleven days; Ortho 10/11) and 481 user cycles of a monophasic contraceptive pill (norethindrone acetate 1500 mcg/ethinyl estradiol 30 mcg daily; Loestrin). The study found no significant differences in intermenstrual bleeding, amenorrhea and study discontinuation due to intermenstrual bleeding between the biphasic and monophasic oral contraceptive pills. REVIEWER'S CONCLUSIONS: Conclusions are limited by the identification of only one trial, the methodological shortcomings of that trial, and the absence of data on accidental pregnancies. However, the trial found no important differences in bleeding patterns between the biphasic and monophasic preparations studied. Since no clear rationale exists for biphasic pills and since extensive evidence is available for monophasic pills, the latter are preferred. PMID- 11406027 TI - Early volume expansion for prevention of morbidity and mortality in very preterm infants. AB - BACKGROUND: Reduced perfusion of organs such as the brain, heart, kidneys and the gastrointestinal tract may lead to acute dysfunction and be associated with permanent injury. Various strategies have been used to provide cardiovascular support to preterm infants including inotropes, corticosteroids and volume expansion. OBJECTIVES: In very preterm infants, does early volume expansion reduce morbidity and mortality. If volume expansion is effective, what type of volume expansion is most effective. SEARCH STRATEGY: The standard search strategy of the Neonatal Review Group was used. Searches were conducted of the Oxford Database of Perinatal Trials, Cochrane Controlled Trials Register, MEDLINE, previous reviews including cross references, abstracts and conference proceedings. SELECTION CRITERIA: Randomised trials of early volume expansion with normal saline, fresh frozen plasma, albumin, plasma substitutes or blood compared to no treatment or another form of volume expansion in preterm infants < 32 weeks gestation or < 1500g were included. Volume expansion was defined as at least 10 mls/kg given in the first 72 hours of life. DATA COLLECTION AND ANALYSIS: Standard methods of the Neonatal Review Group with use of relative risk (RR), risk difference (RD) and weighted mean difference (WMD). The fixed effects model using RevMan 4.1 was used for meta-analysis. Data from individual studies were only eligible for inclusion if a least 80% of infants were reported for that outcome. MAIN RESULTS: Seven studies were included. Five studies, four with data for mortality, compared volume to no treatment. Most studies enrolled very preterm infants on the basis of gestation or birthweight. Two studies comparing different types of volume expansion enrolled very preterm infants with hypotension. No study enrolled infants on the basis of low blood flow. One study examined the effect of volume expansion on blood flow but in normotensive very preterm infants. Comparing volume and no treatment, 4 studies with a total of 940 very preterm infants reported no significant difference in mortality (RR 1.11, 95% CI 0.88, 1.40). The large NNNI 1996 study reported no significant difference in severe disability (RR 0.80, 95% CI 0.52, 1.23), cerebral palsy (RR 0.76, 95% CI 0.48, 1.20) and combined death or severe disability (RR 1.00, 95% CI 0.80, 1.24). Although one small study (Beverley 1985) reported reduced P/IVH with volume expansion, this was not supported by any other study. No significant difference was reported in grade 3-4 P/IVH and combined death or grade 3-4 P/IVH. One study (NNNI 1996) reported no significant difference in the incidence of hypotension. The finding of decreased necrotising enterocolitis and increased sepsis in infants who received fresh frozen plasma compared to a gelatin-based plasma substitute or no treatment in one study should be treated with caution. No significant differences in mortality or disability were found in this study. In one small study, there were no significant differences in outcomes between hypotensive infants who received colloid (albumin) or crystalloid (saline). REVIEWER'S CONCLUSIONS: There is no evidence from randomised trials to support the routine use of early volume expansion in very preterm infants without cardiovascular compromise. There is insufficient evidence to determine whether infants with cardiovascular compromise would benefit from volume expansion. There is insufficient evidence to determine what type of volume expansion should be used in preterm infants (if at all) or for the use of early red cell transfusions. The trial comparing colloid (albumin) and crystalloid (saline) in hypotensive preterm infants found no benefit in using a more expensive blood product compared to saline, but is insufficiently powered to detect an important clinical difference. PMID- 11406028 TI - Early volume expansion versus inotrope for prevention of morbidity and mortality in very preterm infants. AB - BACKGROUND: Reduced perfusion of organs such as the brain, heart, kidneys and the gastrointestinal tract may lead to acute dysfunction and be associated with permanent injury. Various strategies have been used to provide cardiovascular support to preterm infants including inotropes, corticosteroids and volume expansion. OBJECTIVES: In very preterm infants, to determine the effect of early volume expansion compared to inotrope in reducing morbidity and mortality. Subgroup analysis was planned according to method of diagnosis of poor perfusion, postnatal age of treatment and type of volume expansion and inotrope used. SEARCH STRATEGY: The standard search strategy of the Cochrane Neonatal Review Group was used. SELECTION CRITERIA: All randomised trials that compared volume expansion to an inotrope in preterm infants in the first days of life were included. DATA COLLECTION AND ANALYSIS: Data were extracted independently by each author and analysed using the standard methods of the Cochrane Collaboration and its Neonatal Review Group using relative risk (RR), risk difference (RD) and weighted mean difference (WMD). MAIN RESULTS: Two small studies comparing volume expansion, using albumin, with dopamine were included. Both studies were adequately randomised, unblinded studies of albumin versus dopamine with no losses to follow up and analysed by intention to treat. Data for clinical outcomes were available from one study in hypotensive preterm infants in the first day of life. In this study, albumin had a higher failure rate for correcting hypotension (RR 5.2, 95% CI 1.3, 20.6). As 49% of these infants had already been given volume, the question of which treatment should be given first was not answered. A second study compared albumin with dopamine in preterm infants with a normal mean blood pressure at a mean age of 32 hours. Dopamine produced a significant increase in mean blood pressure when compared to infants who received albumin or no treatment, although the difference between the dopamine and albumin groups did not reach significance. Albumin and dopamine produced similar increases in left ventricular output but no significant change in cerebral blood flow. No difference was found in mortality (RR 1.5, 95% CI 0.5, 4.0) or morbidity including any P/IVH, chronic lung disease or retinopathy. There was a higher rate of grade 2-4 P/IVH of borderline statistical significance in infants who received albumin in one study (RR 1.47; 95% CI 0.96 to 2.25: RD 0.27, 95% CI 0.00 to 0.54). No data were available for neurodevelopmental outcomes. REVIEWER'S CONCLUSIONS: Dopamine was more successful than albumin at correcting low blood pressure in hypotensive preterm infants, many of whom had already received volume. Neither intervention has been shown to be superior at improving blood flow, or in improving mortality and morbidity in preterm infants. The trials do not allow any firm conclusions to be made as to whether or when volume or dopamine should be used in preterm infants. PMID- 11406029 TI - Permissive hypercapnia for the prevention of morbidity and mortality in mechanically ventilated newborn infants. AB - BACKGROUND: Experimental animal data and uncontrolled, observational studies in human infants have suggested that hyperventilation and hypocapnia may be associated with increased pulmonary and neurodevelopmental morbidity. Protective ventilatory strategies allowing higher levels of arterial CO2 (permissive hypercapnia) are now widely used in adult critical care. The aggressive pursuit of normocapnia in ventilated newborn infants may contribute to the already present burden of lung disease. However, the safe or ideal range for PCO2 in this vulnerable population has not been established. OBJECTIVES: To assess whether, in mechanically ventilated neonates, a strategy of permissive hypercapnia improves short and long term outcomes (esp. mortality, duration of respiratory support, incidence of chronic lung disease and neurodevelopmental outcome). SEARCH STRATEGY: Standard strategies of the Cochrane Neonatal Review Group were used. Searches were made of the Oxford Database of Perinatal Trials, MEDLINE, CINAHL, and Current Contents. Searches were also made of previous reviews including cross referencing, abstracts, and conference and symposia proceedings published in Pediatric Research. SELECTION CRITERIA: All randomised controlled trials in which a strategy of permissive hypercapnia was compared with conventional strategies aimed at achieving normocapnia (or lower levels of hypercapnia) in newborn infants who are mechanically ventilated were eligible. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Neonatal Review Group were used. Trials identified by the search strategy were independently reviewed by each author and assessed for eligibility and trial quality. Data were extracted separately. Differences were compared and resolved. Additional information was requested from trial authors. Only published data were available for review. Results are expressed as relative risk and risk difference for dichotomous outcomes, and weighted mean difference for continuous variables. MAIN RESULTS: Two trials involving 269 newborn infants were included. Meta-analysis of combined data was possible for three outcomes. There was no evidence that permissive hypercapnia reduced the incidence of death or chronic lung disease at 36 weeks (RR 0.94, 95% CI 0.78, 1.15), intraventricular haemorrhage grade 3 or 4 (RR 0.84, 95% CI 0.54, 1.31) or periventricular leukomalacia (RR 1.02, 95% CI 0.49, 2.12). There were no differences in any other reported outcomes when the strategy of permissive hypercapnia/minimal ventilation was compared to routine ventilation in newborn infants. Long term neurodevelopmental outcomes were not reported. One trial reported that permissive hypercapnia reduced the incidence of chronic lung disease in the 501 to 750 gram subgroup. REVIEWER'S CONCLUSIONS: This review does not demonstrate any significant overall benefit of a permissive hypercapnia/minimal ventilation strategy compared to a routine ventilation strategy. At present, therefore, these ventilation strategies cannot be recommended to reduce mortality, or pulmonary and neurodevelopmental morbidity. Ventilatory strategies which target high levels of PCO2 (> 55 mmHg) should only be undertaken in the context of well-designed controlled clinical trials. These trials should aim to establish the safe, or ideal, range for CO2 in ventilated newborns, and examine the role of protective ventilatory techniques in achieving this target. PMID- 11406030 TI - Intravenous immunoglobulin for Guillain-Barre syndrome. AB - BACKGROUND: Guillain-Barre syndrome is a potentially serious, acute, paralysing, probably autoimmune disease caused by inflammation of the peripheral nerves. Recovery has been shown to be speeded by plasma exchange which replaces the patient's own plasma with a plasma substitute. Intravenous immunoglobulin purified from donated blood is beneficial in other autoimmune diseases and is easier to administer. OBJECTIVES: To determine the efficacy of intravenous immunoglobulin in comparison with no treatment or other treatments for treating Guillain-Barre syndrome and to determine the most efficacious dose. SEARCH STRATEGY: Search of the Cochrane Neuromuscular Disease Group register using Guillain-Barre syndrome and acute polyradiculoneuritis as the search terms, bibliographies of trials and contact with their authors and other experts. SELECTION CRITERIA: Randomised and quasi-randomised trials. DATA COLLECTION AND ANALYSIS: Two reviewers examined the titles and abstracts of all the papers retrieved by the search, extracted the data onto forms designed for this review, and independently assessed the quality of the trials. MAIN RESULTS: The only trial comparing intravenous immunoglobulin with supportive treatment was inadequate to establish its value. Another Cochrane systematic review has shown that plasma exchange (PE) hastens recovery. Plasma exchange has become the gold standard against which other treatments need to be compared. We found three randomised trials that compared intravenous immunoglobulin with PE. We were able to combine the results of the two largest trials in a metaanalysis involving 398 patients. The primary outcome measure in this review was the change in a 7 grade disability scale four weeks after randomisation. The weighted mean difference of this measure was not significant, being only 0.11 (95% CI -0.14 to 0.37) of a disability grade more improvement in the intravenous immunoglobulin group than the PE group. There were also no significant differences in other outcome measures, including time to walk unaided, mortality, and proportion of patients unable to walk without aid after a year but some of these outcome measures were only available for one trial. We also reviewed one trial involving 249 patients which compared PE followed by intravenous immunoglobulin with PE alone and another involving 37 patients which compared immunoabsorption followed by intravenous immunoglobulin with immunoabsorption alone. Neither revealed any significant differences between the regimens with and without intravenous immunoglobulin. We did not discover any dose ranging studies of intravenous immunoglobulin except for one that is ongoing. REVIEWER'S CONCLUSIONS: There are no adequate trials to determine whether intravenous immunoglobulin is more beneficial than placebo. Intravenous immunoglobulin and plasma exchange have a similar ability to speed the recovery from Guillain-Barre syndrome. Giving intravenous immunoglobulin after plasma exchange is not significantly better than plasma exchange alone. Randomised trials are needed to decide whether intravenous immunoglobulin helps in mild Guillain-Barre syndrome or in disease which has lasted more than two weeks. Randomised trials also need to establish the optimal dose. PMID- 11406031 TI - Prompts to encourage appointment attendance for people with serious mental illness. AB - BACKGROUND: Prompts to encourage attendance at clinics are often used in day-to day practice by diligent carers of people with mental health problems. These may take the form of telephone prompting, financial incentives or issuing a copy of the referral letter to the appointee. OBJECTIVES: To estimate the effects of simple prompting by professional carers to encourage attendance at clinics for those with serious mental illness. SEARCH STRATEGY: Methodical searches of Biological Abstracts (1985-2000), CINAHL (1982-2000), Cochrane Schizophrenia Group's Register (June 2000), Cochrane Library (Issue 2, 2000), EMBASE (1980 2000), MEDLINE (1966-2000) and PsycLIT (1887-2000) were undertaken. These were supplemented by searching of reference lists, personal contact and hand searching of high yield journals. SELECTION CRITERIA: All relevant randomised (or quasi randomised) studies comparing the addition of 'prompts' to standard care for those with serious mental illnesses such as schizophrenia. Prompts had the stated purpose of encouraging attendance or contact with mental health teams and could be text-based, electronic, by telephone call, by personal visit, or could employ financial or other rewards. DATA COLLECTION AND ANALYSIS: Studies and data were independently selected and extracted. For homogeneous dichotomous data the random effects relative risk (RR), the 95% confidence intervals (CI) and, where appropriate, the number needed to treat (NNT) were calculated on an intention-to treat basis. For continuous data the reviewers calculated weighted mean differences. MAIN RESULTS: Only three relevant trials were identified (total n=597). It is not clear whether there is any real difference between attendance of those prompted by telephone one or two days before the appointment, and those given the standard appointment management system (2 trials, n=457, RR missed appointment 0.84 CI 0.7 to 1.1). Text-based prompts, a few days before the appointment day, did increase clinic attendance when compared with no prompt (2 trials, n=200, RR missed appointment 0.6 CI 0.4 to 0.9, NNT 6 CI 2 to 14). Only one small study (n=61) reported data on the combination of telephone and text based prompts versus no prompt (RR missed appointments 0.7 CI 0.4 to 1.2). When telephone prompts were compared with text-based prompts (1 trial, n=75), the latter, in the form of an 'orientation statement' (a short paragraph, taking about 30 seconds to read, explaining the programme of care, the fee system, and providing gentle encouragement) may be somewhat more effective than the telephone prompt (RR missed appointments 1.9 CI 0.98 to 3.8). One last study (n=120) compared a standard letter prompt with a letter 'orientation statement'. Overall, results tended to favour the orientation statement approach rather than the simple letter prompting attendance but the results did not reach conventional levels of statistical significance (RR missed appointments 1.6 CI 0.9 to 2.9). REVIEWER'S CONCLUSIONS: There is evidence that a simple prompt to attend clinic, very close to the time of the appointment may encourage attendance, and a simple orientation-type letter, 24 hours before the clinic appointment, may be more effective than a telephone prompt. This simple intervention could be a more cost effective means of encouraging compliance at first attendance, but supplementing these data with the results of large, well designed, conducted and reported randomised studies would be desirable. PMID- 11406032 TI - Speech and language therapy for dysarthria due to non-progressive brain damage. AB - BACKGROUND: Dysarthria is a common sequel of non-progressive brain damage (typically stroke and traumatic brain damage). Impairment-based therapy and a wide variety of compensatory management strategies are undertaken by speech and language therapists with this patient population. OBJECTIVES: To determine the efficacy of speech and language therapy interventions for adults with dysarthria following non-progressive brain damage. SEARCH STRATEGY: This review has drawn on the search strategies developed for the following Cochrane Groups as a whole: Stroke, Injuries, and Infectious Diseases. Relevant trials were identified in the Specialised Registers of Controlled Trials (see Review Group Details for more information). We also searched the trials register of the Cochrane Rehabilitation and Related Therapies Field. The Cochrane Controlled Trials Register, MEDLINE, EMBASE, CINAHL, PsycLIT, and Linguistics and Language Behavior Abstracts were electronically searched. Hand-searching of The International Journal of Language and Communication Disorders and of reference lists from relevant articles and conference proceedings was also undertaken. Colleagues were approached to identify other possible published and unpublished studies. Date of most recent searches: May 2000. SELECTION CRITERIA: Unconfounded randomised controlled trials. DATA COLLECTION AND ANALYSIS: One reviewer assessed trial quality. Two co reviewers were available to examine any potential trials for possible inclusion in the review. MAIN RESULTS: No trials of the required standard were identified. REVIEWER'S CONCLUSIONS: There is no evidence of the quality required by this review to support or refute the effectiveness of Speech and Language Therapy interventions for dysarthria following non-progressive brain damage. There is an urgent need for good quality research in this area. PMID- 11406033 TI - Combined chemotherapy and radiotherapy (without surgery) compared with radiotherapy alone in localized carcinoma of the esophagus. AB - BACKGROUND: Esophageal carcinoma can be managed primarily with either a surgical or radiotherapeutic (non surgical) approach. Strategies to improve the outcome of either modality alone include the use of combined modalities. Combination chemotherapy radiotherapy is one approach that has been explored over the years with increasing application in clinical practice especially in North America. OBJECTIVES: To evaluate the effectiveness of combined chemotherapy and radiotherapy versus radiotherapy alone in the outcome of patients with localized esophageal carcinoma. Outcomes of interest include overall survival, cause specific survival, local recurrence, dysphagia relief, quality of life, acute and chronic toxicities. SEARCH STRATEGY: The Cochrane strategy for identifying randomized trials was combined with MeSH headings including esophageal neoplasms, radiotherapy, chemotherapy combined modality, drug therapy combination. Medline, Cancerlit and Embase were searched using this strategy. In addition, the Cochrane library was also searched. References from relevant articles and personal files were included. SELECTION CRITERIA: Randomized controlled trials in patients with localized esophageal cancer, with one arm employing radiotherapy alone, and one arm employing combination radiotherapy chemotherapy were included. Studies comparing non chemotherapy agents such as pure radiotherapy sensitisers, immunostimulants, planned esophagectomy, were excluded. DATA COLLECTION AND ANALYSIS: Data were extracted by two independent reviewers, and the trial quality was assessed using both the Jadad scoring and Detsky checklist. Sensitivity analysis was planned to explore sources of heterogeneity where heterogeneity existed. The factors hypothesized a priori included combination versus sequential treatment, quality of study, biological effective radiotherapy dose (i.e. Radiotherapy dose) cisplatin versus non cisplatin containing trials, and 5FU versus non 5FU containing trials. Odds Ratio (OR) and 95% confidence limits were used to assess the significance of the difference between the treatment arms. Absolute risk difference and number needed to treat (NNT) were used to express the magnitude of difference where appropriate. MAIN RESULTS: Thirteen randomized trials were included in the analysis. There were eight concomitant and five sequential radiotherapy and chemotherapy (RTCT) studies. The studies were analyzed separately due to observed heterogeneity across all the studies and biological considerations. Concomitant RTCT provided significant overall reduction in mortality at 1 and 2 years. The mortality in the control arms was 67% and 86% respectively. Combined RTCT provided an absolute reduction of mortality by 9% (95% CI 2-17%) and 8% (95% CI 1-17%) respectively. Expressed as NNT, this is 11 and 10 respectively. At longer follow up, the results were heterogeneous, cautioning against pooling of the data. There was a reduction in the overall local recurrence rate. The local recurrence rate for the control arms was in the order of 69%. Combined RTCT provided an absolute reduction of local recurrence rate of 5% (95% CI 4-26%) with a NNT of 7. There was significant increase of severe and life threatening toxicities with a NNH of 6, with this approach. With the sensitivity analysis, there was a suggestion that cisplatin based and 5FU based chemotherapy studies were reasonable regimens to employ when using this strategy. The results from the sequential RTCT studies were heterogeneous and could not be pooled. Factors hypothesized a priori did not identify any single source that could account for a significant component of the heterogeneity. Examining the results individually, there was no data to support clinical benefit. This approach was also accompanied by significant toxicities. REVIEWER'S CONCLUSIONS: When a non-operative approach is selected, then concomitant RTCT is superior to the RT alone. This approach is accompanied by significant toxicities. In patients who are in good general condition, and the risk benefit has been thoroughly discussed with the patient, concomitant RTCT should be considered for the management of esophageal cancer compared with radiotherapy alone. PMID- 11406034 TI - Recombinant FSH versus urinary gonadotrophins or recombinant FSH for ovulation induction in subfertility associated with polycystic ovary syndrome. AB - BACKGROUND: Over the last four decades, various urinary FSH (uFSH) products of different purity have been developed. In 1988 recombinant FSH (rFSH ) was prepared by transfecting Chinese hamster ovary cell lines with both FSH subunit genes. Both rFSH and uFSH are known to be effective in inducing ovulation in women with clomiphene-resistant polycystic ovary syndrome. Ovulation induction with FSH bears the risk of multiple follicle development, multiple pregnancies and ovarian hyperstimulation syndrome. The dose regimen used can affect the incidence of these complications. OBJECTIVES: To compare in women with clomiphene resistant polycystic ovary syndrome (PCOS) the safety and effectiveness in terms of ovulation, pregnancy, miscarriage, multiple pregnancy rate and ovarian hyperstimulation syndrome (OHSS) of 1) rFSH with uFSH and 2) different dose regimens of rFSH. SEARCH STRATEGY: The search strategy of the Menstrual Disorders and Subfertility review group was used to identify all relevant trials. Please see Review Group details. SELECTION CRITERIA: All relevant published RCT's were selected. Randomised controlled trials were eligible for inclusion if treatment consisted of recombinant FSH versus urinary FSH or recombinant FSH in different dose regimens, to induce ovulation in subfertile women with PCOS. DATA COLLECTION AND ANALYSIS: A computerised MEDLINE and EMBASE search was used to identify randomised and non randomised controlled trials. The reference lists of all studies found were checked for relevant articles. Handsearching of bibliographies of relevant publications and reviews and abstracts of scientific meetings was performed. Serono Benelux BV and NV Organon, the manufacturers of follitropin alpha (Gonal F(R)) and follitropin beta (Puregon(R)) respectively, were asked for unpublished data and ongoing studies. Relevant data were extracted independently by two reviewers (NB, MW). Validity was assessed in terms of method of randomisation, completeness of follow-up, presence or absence of cross-over and co-intervention. All trials were screened and analysed according to predetermined quality criteria. DATA SYNTHESIS: 2X2 tables were generated for all the relevant outcomes. Odds ratios were generated using the Peto modified Mantel-Haenszel technique. MAIN RESULTS: Four randomised trials comparing rFSH versus uFSH were identified. No significant differences were demonstrated for the relevant outcomes. The odds ratio for ovulation rate was 1.19 (95% CI 0.78,1.80), for pregnancy rate 0.95 (95% CI 0.64,1.41), for miscarriage rate 1.26 (95% CI 0.59,2.70), for multiple pregnancy rate 0.44 (95% CI 0.16,1.21) and for OHSS 1.55 (95% CI 0.50,4.84). Similarly, in the only randomised trial that compared chronic low dose versus conventional regimen with rFSH no significant differences were found. REVIEWER'S CONCLUSIONS: At this moment there are not sufficient data to determine which of rFSH or uFSH is preferable for ovulation induction in women with PCOS. PMID- 11406035 TI - Palliative radiotherapy regimens for non-small cell lung cancer. AB - BACKGROUND: Palliative radiotherapy (RT) to the chest is often used in patients with lung cancer, but RT regimens are more often based on tradition than research results. OBJECTIVES: To discover the most effective and least toxic regimens of palliative RT, and whether higher doses increase survival. SEARCH STRATEGY: Electronic databases, reference lists, handsearching of journals and conference proceedings, and discussion with experts were used to identify potentially eligible trials, published and unpublished. SELECTION CRITERIA: Randomised controlled clinical trials comparing different regimens of palliative RT in patients with non-small lung cancer. DATA COLLECTION AND ANALYSIS: Ten randomised trials were reviewed. There were important differences in the doses of RT investigated, the patient characteristics and the outcome measures. Because of this heterogeneity no meta-analysis was attempted. MAIN RESULTS: There is no strong evidence that any regimen gives greater palliation. Higher dose regimens give more acute toxicity. There is evidence for a modest increase in survival (6% at 1 year and 3% at 2 years) in patients with better performance status (PS) given higher dose RT. REVIEWER'S CONCLUSIONS: The majority of patients should be treated with short courses of palliative RT, of 1 or 2 fractions. Care should be taken with the dose to the spinal cord. The use of high dose palliative regimens should be considered for and discussed with selected patients with good PS. More research is needed into reducing the acute toxicity of large fraction regimens and into the role of radical compared to high dose palliative RT and more homogeneous studies are needed. PMID- 11406036 TI - Hydroxyurea for sickle cell disease. AB - BACKGROUND: Sickle cell disease is one of the most common inherited diseases world wide. It is associated with life long morbidity and a reduced life expectancy. Hydroxyurea, a chemotherapeutic drug taken by mouth, raises fetal haemoglobin and, as such, is expected to ameliorate some of the clinical problems of sickle cell disease. OBJECTIVES: To assess the effects of hydroxyurea therapy in sickle cell disease patients of all types, of any age, regardless of setting. SEARCH STRATEGY: We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group specialised register of controlled trials for haemoglobinopathies, which comprises references identified from comprehensive electronic database searches, hand-searching relevant journals, and hand-searching abstract books of conference proceedings. Date of the most recent search(es): November 2000. SELECTION CRITERIA: All randomised or quasi-randomised controlled trials comparing the use of oral hydroxyurea for one month or longer with placebo, standard therapy or other interventions for the treatment of patients with sickle cell disease. DATA COLLECTION AND ANALYSIS: Both reviewers independently assessed trial quality and extracted data from the two studies included. MAIN RESULTS: Twenty trials were found of which two trials, which reported results from a total of 324 adults and children were suitable for inclusion in the review. From the data provided in the published reports only one study (the MSH study to the United States of America) could be analysed. This study showed marked differences in favour of hydroxyurea treatment as compared with placebo in terms of annual crisis rate, use of transfusions, and life-threatening complications (in particular, the acute sickle chest syndrome). No serious adverse effects were reported from either study. REVIEWER'S CONCLUSIONS: While hydroxyurea appears both effective and safe in the severely affected SS adults over a two year period; further studies are required to elucidate its role in other patient groups and for other conditions. PMID- 11406037 TI - Media-based behavioural treatments for behavioural disorders in children. AB - BACKGROUND: Prevalence studies show that behaviour problems in children are quite common (10-15% in preschoolers). These problems may manifest as, for example anxiety, sadness, over-activity and tantrums. Some studies have shown that these problems can be persistent, and that they lead to a range of problems in adolescence and adulthood. Many approaches are used to address behavioural problems such as medication, or more usually, psychological treatments either directly with the child and/or his/her family. Behavioural interventions have been shown to be highly effective but access to these treatments is limited due to factors such as time and expense. Presenting the information parents need in order to manage these behaviour problems in booklet or other media-based format would reduce the cost and thus increase access to these interventions. In the adult population it seems that media-based interventions such as these can be moderately effective. Given that the cost of media-based treatment is so low it is useful to know how effective they are when given to parents. It was hypothesised that media-based treatments would be less effective than conventional psychological treatments and that efficacy would improve with increasing amounts of therapist intervention. OBJECTIVES: To review the effects of media-based behavioural therapies (definitions below), for any young person with a behavioural disorder (however diagnosed) compared to standard care and no treatment controls. SEARCH STRATEGY: A range of electronic databases were systematically searched using a specified search strategy. Individual journals of interest were hand-searched where necessary, references in all selected trials were checked for other trials and, where it was thought to be of possible use, individual authors were contacted for additional information. SELECTION CRITERIA: Randomised controlled trials of behavioural media-based treatments for behaviour problems in children. DATA COLLECTION AND ANALYSIS: Abstracts and titles of studies identified from searches of electronic databases were read to determine whether they met the inclusion criteria. Full copies of those possibly meeting these criteria from electronic or other searches were assessed by the reviewer and queries were resolved by discussion with an independent rater. Data were analysed using Revman. MAIN RESULTS: In general, media-based therapies for behavioural disorders in children had a moderate effect when compared with both no-treatment controls and with standard care. Significant improvements were often made with the addition of up to 2 hours of therapist time. REVIEWER'S CONCLUSIONS: These formats of delivering behavioural interventions for carers of children are possibly worth considering in clinical practice. For straightforward cases media-based interventions may be enough to make clinically significant changes in a child's behaviour, and reduce the amount of time primary care workers have to devote to each case. Consequently this would increase the number of families who could possibly benefit from this type of intervention. Media based therapies would therefore appear to have both clinical and economic implications as regards the treatment of children with behavioural problems. PMID- 11406038 TI - Chinese medicinal herbs for asymptomatic carriers of hepatitis B virus infection. AB - BACKGROUND: About 350 million people are chronically infected carriers of hepatitis B virus and are at a higher risk of serious illness and death from cirrhosis of the liver and liver cancer. Chinese medicinal herbs have been used widely for more than 2000 years to treat chronic liver disease. OBJECTIVES: To assess whether Chinese medicinal herbs are effective and safe for treating asymptomatic carriers of hepatitis B virus. SEARCH STRATEGY: The trials registers of the Cochrane Hepato-Biliary Group, the Cochrane Library and the Cochrane Complementary Medicine Field were searched in combination with MEDLINE, EMBASE, and handsearches of Chinese journals and conference proceedings. SELECTION CRITERIA: Randomised or quasi-randomised trials (minimum follow-up three months) in asymptomatic carriers of hepatitis B virus. Chinese medicinal herbs (single herb or compound of herbs) compared with placebo, no intervention, general non specific treatment, or interferon treatment. Trials of Chinese medicinal herbs plus interferon versus interferon alone were also included. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. Analysis was performed by intention-to-treat where possible. Pre-specified subgroup analyses were: ethnic origin, age at time of infection, and single herb or compound of herbs. MAIN RESULTS: Three randomised clinical trials (307 patients) that followed patients for three months or more after the end of treatment were included. The methodological quality was poor. The herbal compound 'Jianpi Wenshen recipe' had significant effects on viral markers compared to interferon: relative risk 2.40 (95% CI 1.01 to 5.72) for clearance of serum HBsAg, 2.03 (95% CI 0.98 to 4.20) for clearance of HBeAg, and 2.54 (95% CI 1.13 to 5.70) for seroconversion of HBeAg to anti-HBe. Phyllanthus amarus and Astragalus membranaceus showed no significant antiviral effect compared with placebo. Analysis of pooling eight randomised clinical trials with less than three months follow-up did not show a significant benefit of Chinese medicinal herbs on viral markers. Data on long-term clinical outcomes and quality of life were lacking. REVIEWER'S CONCLUSIONS: Based on one low quality trial, the medicinal herb 'Jianpi Wenshen recipe' may have an antiviral activity in asymptomatic carriers of hepatitis B virus. However, rigorous randomised, double-blind, placebo controlled trials are needed before herbs should be used for this condition. PMID- 11406039 TI - Interventions for promoting smoke alarm ownership and function. AB - BACKGROUND: Residential fires caused at least 67 deaths and 2,500 non-fatal injuries to children aged 0-16 in the United Kingdom in 1998. Smoke alarm ownership is associated with a reduced risk of residential fire death. OBJECTIVES: We evaluated interventions to promote residential smoke alarms, to assess their effect on smoke alarm ownership, smoke alarm function, fires and burns and other fire-related injuries. SEARCH STRATEGY: We searched the Cochrane Controlled Trials Register, Cochrane Injuries Group database, MEDLINE, EMBASE, PsycLIT, CINAHL, ERIC, Dissertation Abstracts, International Bibliography of Social Sciences, ISTP, FIREDOC and LRC. Conference proceedings, published case studies, and bibliographies were systematically searched, and investigators and relevant organisations were contacted, to identify trials. SELECTION CRITERIA: Randomised, quasi-randomised or nonrandomised controlled trials completed or published after 1969 evaluating an intervention to promote residential smoke alarms. DATA COLLECTION AND ANALYSIS: Two reviewers independently extracted data and assessed trial quality. MAIN RESULTS: We identified 26 trials, of which 13 were randomised. Overall, counselling and educational interventions had only a modest effect on the likelihood of owning an alarm (OR=1.26; 95% CI: 0.87 to 1.82) or having a functional alarm (OR=1.19; 0.85 to 1.66). Counselling as part of primary care child health surveillance had greater effects on ownership (OR=1.96; 1.03 to 3.72) and function (OR=1.72; 0.78 to 3.80). Results were sensitive to trial quality, however, and effects on fire-related injuries were not reported. In two non randomised trials, direct provision of free alarms significantly increased functioning alarms and reduced fire-related injuries. Media and community education showed little benefit in non randomised trials. REVIEWER'S CONCLUSIONS: Counselling as part of child health surveillance may increase smoke alarm ownership and function, but its effects on injuries are unevaluated. Community smoke alarm give-away programmes apparently reduce fire related injuries, but these trials were not randomised and results must be interpreted cautiously. Further efforts to promote smoke alarms in primary care or through give-away programmes should be evaluated by adequately designed randomised controlled trials measuring injury outcomes. PMID- 11406040 TI - Antihypertensive drug therapy for mild to moderate hypertension during pregnancy. AB - BACKGROUND: Mild-moderate hypertension during pregnancy is common. Antihypertensive drugs are often used in the belief that lowering blood pressure will prevent progression to more severe disease, and thereby improve outcome. OBJECTIVES: To assess the effects of antihypertensive drug treatments for women with mild to moderate hypertension during pregnancy. SEARCH STRATEGY: Relevant trials were identified in the register of trials maintained by the Cochrane Pregnancy and Childbirth Group. In addition, the Cochrane Controlled Trial Register, MEDLINE, and EMBASE were searched. Date of last search: October 2000. SELECTION CRITERIA: All randomised trials evaluating any antihypertensive drug treatment for mild to moderate hypertension during pregnancy, defined whenever possible as systolic blood pressure 140-169 mmHg and diastolic blood pressure 90 109 mmHg. Comparisons were of one or more antihypertensive drug(s) with placebo, with no antihypertensive drug, or with another antihypertensive drug, and where treatment was planned to continue for at least seven days. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers. MAIN RESULTS: Overall, this review includes 40 studies (3797 women), 24 of which compared an antihypertensive drug with placebo/no antihypertensive drug (2815 women). There is a halving in the risk of developing severe hypertension associated with the use of antihypertensive drug(s) [17 trials, 2155 women; relative risk (RR) 0.52 (95% confidence interval (CI) 0.41 to 0.64); risk difference (RD) -0.09 (-0.12 to -0.06); number needed to treat (NNT) 12 (9 to 17)] but little evidence of a difference in the risk of pre-eclampsia [19 trials, 2402 women; RR 0.99 (0.84 to 1.18)]. Similarly, there is no clear effect on the risk of the baby dying [23 trials, 2727 women; RR 0.71(0.46 to 1.09)], preterm birth [12 trials, 1738 women; RR 0.98 (0.85 to 1.13)], or small for gestational age babies [17 trials, 2159 women; RR 1.13 (0.91 to 1.42)]. There were no clear differences in any other outcomes. Seventeen trials (1182 women) compared one antihypertensive drug with another. There is no clear difference between any of these drugs in the risk of developing severe hypertension, and proteinuria/pre-eclampsia. Other antihypertensive agents seem better than methyldopa for reducing the risk of the baby dying [14 trials, 1010 subjects, RR 0.49 (0.24 to 0.99); RD -0.02 (-0.04 to 0.00); NNT 45 (22 to 1341)]. Other outcomes were only reported by a small proportion of studies, and there were no clear differences. REVIEWER'S CONCLUSIONS: It remains unclear whether antihypertensive drug therapy for mild moderate hypertension during pregnancy is worthwhile. PMID- 11406041 TI - Diuretic therapy for newborn infants with posthemorrhagic ventricular dilatation. AB - BACKGROUND: Intraventricular hemorrhage remains a serious complication of premature birth and post hemorrhagic hydrocephalus still has no satisfactory treatment. Acetazolamide and furosemide, which both reduce the production of cerebrospinal fluid, have been suggested as non-invasive therapies to reduce hydrocephalus and the need for ventriculo-peritoneal (V-P) shunting. OBJECTIVES: The aim of this review was to determine whether the use of acetazolamide and furosemide improves outcome, especially shunt dependence, in infants developing post-hemorrhagic ventricular dilatation. SEARCH STRATEGY: The standard search strategy of the Cochrane Collaboration was used. SELECTION CRITERIA: Randomised, or quasi-randomised trials, of acetazolamide and/or furosemide compared with standard therapy in infants with IVH or post-hemorrhagic ventricular dilatation DATA COLLECTION AND ANALYSIS: Data were extracted independently by each author and were analysed by the standard methods of the Cochrane Collaboration using relative risk (RR) and risk difference (RD), a fixed effect model and sensitivity analyses where appropriate. MAIN RESULTS: There were two eligible trials: one randomized 16 infants and the other, 177. Neither showed a decreased risk for V-P shunt or for V-P shunt or death associated with acetazolamide and furosemide therapy. The larger trial showed that acetazolamide and furosemide treatment resulted in a borderline increase in the risk for motor impairment at one year (RR 1.27, CI 1.02 to 1.58; RD 0.16, CI 0.02 to 0.31), but did not significantly affect the risk for the combined outcome of delay, disability or motor impairment among survivors, or the risk of the combined outcome of death, delay, disability or impairment at one year. The larger trial showed that diuretic treatment increased the risk for nephrocalcinosis (RR 5.31, CI 1.90 to 14.84; RD 0.19, CI 0.09 to 0.29); meta-analysis confirmed this result. REVIEWER'S CONCLUSIONS: Acetazolamide and furosemide therapy is neither effective nor safe in treating post hemorrhagic ventricular dilatation. Acetazolamide and furosemide cannot be recommended as therapy for post hemorrhagic hydrocephalus. PMID- 11406042 TI - Effects of nonsteroidal anti-inflammatory drugs on post-operative renal function in normal adults. AB - BACKGROUND: Nonsteroidal anti-inflammatory drugs (NSAIDs) can play a major role in the management of acute pain in the peri-operative period. However, there there are conflicting views on whether NSAIDs are associated with adverse renal effects. OBJECTIVES: The primary objective of this review was to determine the effects of NSAIDs on post-operative renal function in adults with normal pre operative renal function. SEARCH STRATEGY: Electronic searches for relevant randomised and quasi-randomised controlled trials in Cochrane Controlled Trials Register, MEDLINE and EMBASE were performed. Attempts were also made to identify trials from citation lists of relevant trials, review articles and clinical practice guidelines. Hand-searching of conference abstracts published in major anaesthetic journals was also performed. SELECTION CRITERIA: The inclusion criteria were randomised or quasi-randomised comparisons of individual NSAIDs with either each other or placebo for treatment of post-operative pain, with relevant post-operative renal outcome measures, in adult surgical patients with normal renal function. DATA COLLECTION AND ANALYSIS: Of the 14 trials that fulfilled the selection criteria for this review, eight trials were relevant with sufficient data for meta-analysis. The data was extracted independently by two reviewers. The primary outcome measure was creatinine clearance within the first two days after surgery. Secondary outcome measures included serum creatinine, urine volume, urinary sodium level, urinary potassium level, fractional excretion of sodium, fractional excretion of potassium, need for dialysis and need for diuretic or dopamine treatment for renal insufficiency. Weighted mean differences for continuous outcomes and relative risk for dichotomous outcomes were estimated. MAIN RESULTS: As a group, NSAIDs reduced creatinine clearance by 18ml/min (95%CI: 6 to 31) and potassium output by 38mmol/day (95%CI: 19 to 56) on the first day after surgery compared to placebo. Serum creatinine clearance increased on the second day after surgery by 15umol/L (95%CI: 2 to 28) compared to placebo. No significant reduction in urine volume during the early post operative period was found. There was no significant difference in serum creatinine in the early post-operative period between patients receiving ketorolac and diclofenac in one trial. No cases of post-operative renal failure requiring dialysis were described. REVIEWER'S CONCLUSIONS: NSAIDs caused a clinically unimportant transient reduction in renal function in the early post operative period in patients with normal pre-operative renal function. NSAIDs should not be withheld from adults with normal pre-operative renal function because of concerns about post-operative renal impairment. PMID- 11406043 TI - Spinal immobilisation for trauma patients. AB - BACKGROUND: Spinal immobilisation involves the use of a number of devices and strategies to stabilise the spinal column after injury and thus prevent spinal cord damage. The practice is widely recommended and widely used in trauma patients with suspected spinal cord injury in the pre-hospital setting. OBJECTIVES: To quantify the effect of different methods of spinal immobilisation (including immobilisation versus no immobilisation) on mortality, neurological disability, spinal stability and adverse effects in trauma patients. SEARCH STRATEGY: We searched the Cochrane Controlled Trial Register (CCTR), the specialised register of the Cochrane Injuries Group, MEDLINE, EMBASE, CINAHL, PubMed and the National Research Register. We checked reference lists of all articles and contacted experts in the field to identify eligible trials. Manufacturers of spinal immobilisation devices were also contacted for information. SELECTION CRITERIA: Randomised controlled trials comparing spinal immobilisation strategies in trauma patients with suspected spinal cord injury. Trials in healthy volunteers were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied eligibility criteria to trial reports and extracted data. MAIN RESULTS: We found no randomised controlled trials of spinal immobilisation strategies in trauma patients. REVIEWER'S CONCLUSIONS: We did not find any randomised controlled trials that met the inclusion criteria. The effect of spinal immobilisation on mortality, neurological injury, spinal stability and adverse effects in trauma patients remains uncertain. Because airway obstruction is a major cause of preventable death in trauma patients, and spinal immobilisation, particularly of the cervical spine, can contribute to airway compromise, the possibility that immobilisation may increase mortality and morbidity cannot be excluded. Large prospective studies are needed to validate the decision criteria for spinal immobilisation in trauma patients with high risk of spinal injury. Randomised controlled trials in trauma patients are required to establish the relative effectiveness of alternative strategies for spinal immobilisation. PMID- 11406044 TI - Speech and language therapy for dysarthria in Parkinson's disease. AB - BACKGROUND: Dysarthria is a common manifestation of Parkinson's disease which increases in frequency and intensity with the progress of the disease (Streifler 1984). Up to 20% of Parkinsonian patients are referred for speech and language therapy (S & L T), its aim being to improve the intelligibility of the patient's speech. OBJECTIVES: To compare the efficacy of speech and language therapy versus placebo or no interventions in patients with Parkinson's disease. SEARCH STRATEGY: Relevant trials were identified by electronic searches of MEDLINE, EMBASE, CINAHL, ISI-SCI, AMED, MANTIS, REHABDATA, REHADAT, GEROLIT, Pascal, LILACS, MedCarib, JICST-EPlus, AIM, IMEMR, SIGLE, ISI-ISTP, DISSABS, Conference Papers Index, Aslib Index to Theses, the Cochrane Controlled Trials Register, the CentreWatch Clinical Trials listing service, the metaRegister of Controlled Trials, ClinicalTrials.gov, CRISP, PEDro, NIDRR and NRR; and examination of the reference lists of identified studies and other reviews. SELECTION CRITERIA: Only randomised controlled trials (RCT) were included. DATA COLLECTION AND ANALYSIS: Data were abstracted independently by KD and RW and differences settled by discussion. MAIN RESULTS: Three randomised controlled trials were found comparing speech and language therapy with placebo for speech disorders in Parkinson's disease. A total of 63 patients were examined. The loudness of the patients' voices were increased by between 7-18%, depending on the speaking task being performed. It is likely that this is a clinically significant improvement. After six months the degree of improvement was reduced but was still statistically significant. Overall measures of dysarthria were measured in two trials and also improved. The clinical significance of these improvements was less clear cut as intelligibility of speech was not measured in any of these studies. REVIEWER'S CONCLUSIONS: Considering the small number of patients examined, the methodological flaws in many of the studies, and the possibility of publication bias, there is insufficient evidence to support or refute the efficacy of speech and language therapy for dysarthria in Parkinson's disease. A Delphi-style survey is needed to develop a consensus as to what is 'standard' S< for dysarthria in Parkinson's disease. Then a large well designed placebo-controlled RCT is needed to demonstrate speech and language therapy's effectiveness for dysarthria in Parkinson's disease. The trial should conform to CONSORT guidelines. Outcome measures with particular relevance to patients should be chosen and the patients followed for at least 6 months to determine the duration of any improvement. PMID- 11406045 TI - A comparison of speech and language therapy techniques for dysarthria in Parkinson's disease. AB - BACKGROUND: Dysarthria is a common manifestation of Parkinson's disease that increases in frequency and intensity with the progress of the disease (Streifler 1984). Up to 20% of Parkinsonian patients are referred for speech and language therapy (S & LT), its aim being to improve the intelligibility of the patient's speech. OBJECTIVES: To compare the efficacy and effectiveness of novel S & LT techniques versus standard S & LT to treat dysarthria in patients with Parkinson's disease. To compare the efficacy and effectiveness of one S & LT technique versus a second form of S & LT to treat Parkinsonian dysarthria. SEARCH STRATEGY: Relevant trials were identified by electronic searches of MEDLINE, EMBASE, CINAHL, ISI-SCI, AMED, MANTIS, REHABDATA, REHADAT, GEROLIT, Pascal, LILACS, MedCarib, JICST-EPlus, AIM, IMEMR, SIGLE, ISI-ISTP, DISSABS, Conference Papers Index, Aslib Index to Theses, the Cochrane Controlled Trials Register, the CentreWatch Clinical Trials listing service, the metaRegister of Controlled Trials, ClinicalTrials.gov, CRISP, PEDro, NIDRR and NRR; and examination of the reference lists of identified studies and other reviews. SELECTION CRITERIA: Only randomised controlled trials (RCT) were included. DATA COLLECTION AND ANALYSIS: Data was abstracted independently by KD and RW and differences settled by discussion. MAIN RESULTS: Only two trials were identified with only 71 patients. The method of randomisation was good in only one trial and the concealment of allocation was inadequate in both trials. These methodological problems could potentially lead to bias from a number of sources. The methods used in the two studies varied so much that meta-analysis of the results was not possible. Scott 83 compared prosodic exercises with visual cues with prosodic exercises alone (See Glossary: Table 01). The authors examined prosody and intelligibility as outcome measures immediately after therapy. Ramig 95 compared the Lee Silverman Voice Therapy (LSVT) which emphasises increased vocal effort, with respiratory therapy which aimed to increase respiratory muscle activity. Ramig 95 examined a wide range of vocal characteristics, activities of daily living affected by speech, depression and the carer's impressions of the patient's speech quality. Some of these outcomes were measured up to 24 months after the end of the therapy. However, in neither study were changes in outcomes due to 'Therapy A' compared with the changes due to 'Therapy B' statistically. Therefore no comment on the comparative efficacy of these types of speech and language therapy can be made. REVIEWER'S CONCLUSIONS: Considering the methodological flaws in both of these studies, the small number of patients examined, and the possibility of publication bias, there is insufficient evidence to support or refute the efficacy of any given form of S & LT over another to treat dysarthria in Parkinson's disease. Given the lack of evidence from RCTs to support or refute the efficacy of S & LT in Parkinson's disease (see Cochrane review 'Speech and Language therapy for Dysarthria in Patients with Parkinson's Disease'), the consensus as to what is considered 'best-practice' S & LT must be proved first through a large well-designed placebo-controlled randomised trial before examining variations in S & LT methodology. The design of these trials should minimise bias and be reported fully using CONSORT guidelines (CONSORT 1996). Outcome measures with particular relevance to patients, their carers, physicians and speech and language therapists should be chosen and the patients followed for at least 6 months to determine the duration of any improvement. PMID- 11406046 TI - Thermotherapy for treating rheumatoid arthritis. AB - BACKGROUND: Heat and cold therapy are often used as adjuncts in the treatment of rheumatoid arthritis by rehabilitation specialists. OBJECTIVES: To evaluate the effects of heat and cold on objective and subjective measures of disease activity in patients with rheumatoid arthritis. SEARCH STRATEGY: We searched Medline, Embase, PEDro, Current Contents, Sports Discus and CINAHL up to June 2000. The Cochrane Field of Rehabilitation and related therapies and the Cochrane Musculoskeletal Review Group were also contacted for a search of their specialized registers. Handsearching was conducted on all retrieved articles for additional articles. SELECTION CRITERIA: Randomized or controlled clinical trials of ice or heat compared to placebo or active interventions in patients with rheumatoid arthritis and case-control and cohort studies were eligible. No language restrictions were applied. Abstracts were accepted. DATA COLLECTION AND ANALYSIS: Two independent reviewers identified potential articles from the literature search. These reviewers extracted data using pre-defined extraction forms. Consensus was reached on all data extraction. Quality was assessed by two reviewers using a 5 point scale that measured the quality of randomization, double-blinding and description of withdrawals. MAIN RESULTS: Three studies (79 subjects) met the inclusion criteria. There was no effect on objective measures of disease activity (including inflammation, pain and x-ray measured joint destruction) of either ice versus control or heat versus control. Patients reported that they preferred heat therapy to no therapy (94% prefer heat therapy to no therapy). There was no difference in patient preference for heat or ice. No harmful effects of ice or heat were reported. REVIEWER'S CONCLUSIONS: Since patients preferred thermotherapy to no therapy, thermotherapy can be used as a palliative therapy which can be applied at home as needed to relieve pain. These results are limited by the poor methodological quality of the trials. PMID- 11406047 TI - Droperidol for acute psychosis. AB - BACKGROUND: People with acute psychotic illnesses, especially when associated with agitated or violent behaviour, may require urgent pharmacological tranquillisation or sedation. Droperidol, a butyrophenone neuroleptic, is used for this purpose in several countries. OBJECTIVES: To estimate the effects of droperidol when compared to other treatments for controlling disturbed behaviour and reducing psychotic symptoms for people with suspected acute psychotic illnesses. SEARCH STRATEGY: The Cochrane Controlled Trials Register (Issue 2, 2000), The Cochrane Schizophrenia Group's Register (May 2000), EMBASE (1980 2000), MEDLINE (1966-2000), PASCAL (1973-2000) and PsycLIT (1970-2000) were methodically searched. Twenty-one other databases were also searched as part of a broader project and this composite database was searched for this review. This was supplemented by hand searching reference lists, contacting industry and relevant authors. SELECTION CRITERIA: Randomised clinical trials comparing droperidol to any treatment, for people with suspected acute psychotic illnesses, such as schizophrenia, schizoaffective disorder, mixed affective disorders, manic phase of bipolar disorder or brief psychotic episode. DATA COLLECTION AND ANALYSIS: Studies were reliably selected, quality assessed and data extracted. Data were excluded where more than 50% of participants were lost to follow up. For binary outcomes, standard estimations of risk ratio (RR) and their 95% confidence intervals (CI) were calculated. Where possible, weighted number needed to treat or harm statistics (NNT, NNH), and their 95% confidence intervals (CI), were also calculated. MAIN RESULTS: Only two clearly relevant randomised trials with usable data were identified. One additional study was included but focused on outcomes at 30 days rather than a few hours. One small (n=41) randomised trial compared droperidol (10mg IV) with placebo IV and found that people allocated to droperidol were significantly less likely to need additional haloperidol injections in the first few minutes (n=41, RR 0.37 CI 0.2 to 0.7, NNT 2 CI 1 to 10) than those given placebo. By 90 minutes this difference was still evident but not statistically significant (RR 0.46 CI 0.2 to 1.2). When 5mg IM droperidol was compared to 5mg IM haloperidol people given droperidol were again less likely to need additional injections by 30 minutes, than those given haloperidol, but this result did not quite reach conventional levels of statistical significance (n=27, RR 0.45 CI 0.2 to 1.01). One person out of 16 given haloperidol experienced a mild dystonic reaction, and none of the 11 people allocated to droperidol were reported to have experienced adverse effects. REVIEWER'S CONCLUSIONS: This is an important and surprisingly under-researched area. Use of droperidol for the emergency situation is currently justified on experience rather than evidence from well conducted and reported randomised trials. PMID- 11406048 TI - Enteral versus parenteral nutrition for acute pancreatitis. AB - BACKGROUND: Acute pancreatitis creates a catabolic stress state promoting a systemic inflammatory response and nutritional deterioration. Adequate supply of nutrients plays an important role to ensure optimum recovery. Total parenteral nutrition (TPN) has been the standard practice for providing exogenous nutrients to patients with severe acute pancreatitis. However, recent data suggest that enteral nutrition (EN) is feasible. Thus, a comparison of EN and TPN in patients with acute pancreatitis needs to be made. OBJECTIVES: To compare the effect of total parenteral nutrition (TPN) versus enteral nutrition (EN) on mortality, morbidity and length of hospital stay in patient with acute pancreatitis. SEARCH STRATEGY: Trials were identified by computerized searches of The Cochrane Controlled Trials Register, MEDLINE, and EMBASE. Additional studies were identified and included where relevant by searching Scisearch, the bibliographies of review articles and identified trials, and personal files. The search was undertaken in August, 2000. No language restrictions were applied. SELECTION CRITERIA: Randomized clinical trials, in which nutrition support with TPN were compared to EN in patients with acute pancreatitis. DATA COLLECTION AND ANALYSIS: Two reviewers independently abstracted data and assessed trial quality. Information was collected on death, length of hospital stay, systemic infection, local septic complications, and other local complications. MAIN RESULTS: Two trials with a total of 70 participants were included. The relative risk (RR) for death with EN vs TPN was 0.56 (95% CI 0.05 to 5.62). Mean length of hospital stay was reduced with EN (WMD -2.20, 95% CI -3.62 to -0.78). RR for systemic infection with EN vs TPN was 0.61 (95% CI 0.29 to 1.28). In one trial, RR for local septic complications and other local complications with EN vs TPN was 0.56 (95% CI 0.12 to 2.68) and 0.16 (95% CI 0.01 to 2.86) respectively. REVIEWER'S CONCLUSIONS: Although there is a trend towards reductions in the adverse outcomes of acute pancreatitis after administration of EN, clearly there are insufficient data to draw firm conclusions about the effectiveness and safety of EN versus TPN. Further trials are required with sufficient size to account for clinical heterogeneity and to measure all relevant outcomes. PMID- 11406049 TI - Reduced osmolarity oral rehydration solution for treating dehydration caused by acute diarrhoea in children. AB - BACKGROUND: Oral rehydration solution (ORS) has reduced childhood deaths from diarrhoea in many countries. Recent studies suggest that the currently recommended formulation of ORS recommended by the World Health Organization (WHO) may not be optimal, and solutions that contain lower concentrations of sodium and glucose may be more effective. OBJECTIVES: In children with acute diarrhoea, to compare reduced osmolarity glucose-based oral rehydration salt solution with international WHO formulation. SEARCH STRATEGY: The Cochrane Collaboration Trials Register, MEDLINE, and EMBASE were searched. Additional trials were identified by hand searching. Content experts were contacted. SELECTION CRITERIA: Randomised controlled trials comparing reduced osmolarity ORS solution with the WHO formulation. Outcomes sought were unscheduled intravenous fluid infusion therapy and measures of clinical illness. DATA COLLECTION AND ANALYSIS: Data were extracted by two reviewers. We tested for heterogeneity using the chi-square statistic, conducted sensitivity analysis by allocation concealment, and the regression approach to assess funnel plot asymmetry from selective trial publication. MAIN RESULTS: The primary outcome was reported in 12 trials. In a meta-analysis of nine trials, reduced osmolarity ORS was associated with fewer unscheduled infusions compared with standard WHO ORS (Mantel Haenzel odds ratio 0.61, 95% confidence interval 0.47 to 0.81) with no evidence for heterogeneity between trials. No unscheduled intravenous fluid infusion therapy was required in any participant in three trials. Thirteen trials reported stool output, and data suggested less stool output in the reduced osmolarity ORS group. Vomiting was less frequent in the reduced osmolarity group in the six trials reporting this. Six trials sought hyponatraemia, with events in three studies, but no obvious difference between the two arms. REVIEWER'S CONCLUSIONS: In children admitted to hospital with diarrhoea, reduced osmolarity ORS when compared to WHO ORS is associated with fewer unscheduled intravenous infusions, smaller stool volume post randomisation, and less vomiting. No additional risk of developing hyponatraemia when compared with WHO ORS was detected. PMID- 11406050 TI - Telephone counselling for smoking cessation. AB - BACKGROUND: Telephone services can provide information and support for smokers. Counselling may be provided proactively or offered reactively to callers to smoking cessation helplines. OBJECTIVES: To evaluate the effect of proactive and reactive telephone support to help smokers quit. SEARCH STRATEGY: We searched the Cochrane Tobacco Addiction Group trials register for studies using free text term 'telephone*' or the keywords 'telephone counselling' or 'Hotlines' or 'Telephone'. Date of the most recent search: August 2000. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials in which proactive or reactive telephone counselling to assist smoking cessation was offered to smokers or recent quitters. DATA COLLECTION AND ANALYSIS: Trials were identified and data extracted by one person and checked by a second. The main outcome measure was abstinence from smoking after at least six months follow-up. We used the most rigorous definition of abstinence in each trial, and biochemically validated rates where available. Participants lost to follow-up were considered to be continuing smokers. Where interventions were similar, we performed meta-analysis using a fixed effects model to give an odds ratio. MAIN RESULTS: Twenty three trials met inclusion criteria. Ten trials compared proactive counselling to a minimal intervention control. There was statistical heterogeneity, with three trials showing a significant benefit, and seven showing non significant differences. Four trials adding telephone support to a face to face intervention control failed to detect a significant effect on long term quit rates. Four trials failed to detect an additional effect of telephone support in users of nicotine replacement therapy. Providing access to a hotline showed a significant benefit in one trial and no significant difference in two. Varying the type of counselling provided has not been shown to affect outcome. REVIEWER'S CONCLUSIONS: Proactive telephone counselling can be effective compared to an intervention without personal contact. There was heterogeneity between trials so the size of effect is uncertain. The available evidence neither confirms nor rules out a benefit of telephone counselling as an adjunct to face to face counselling or pharmacotherapy. Further trials randomising access to helplines are unlikely to be done but indirect evidence suggests they can be a useful part of a smoking cessation service. PMID- 11406051 TI - Radical radiotherapy for stage I/II non-small cell lung cancer in patients not sufficiently fit for or declining surgery (medically inoperable). AB - BACKGROUND: In general, surgery is believed to offer the best prospects for cure for early stage non-small cell lung cancer (NSCLC). In spite of the intention to consider all patients with stage I-II disease for surgery, there are those who, although technically operable, either refuse surgery or are considered inoperable because of insufficient respiratory reserve, cardiovascular disease or general frailty. This group may therefore be considered "medically inoperable". Some respiratory physicians refer these patients for radical radiotherapy whilst others believe that radiotherapy has little to offer and adopt a watch policy, referring patients for palliative radiotherapy only when they become symptomatic. Although there is little evidence from randomised trials to support the use of radical radiotherapy for stage I/II NSCLC, it is the perception of most clinical oncologists (radiotherapists) that patients should receive radical, as opposed to palliative, treatment (COIN 1999). OBJECTIVES: To determine the effectiveness and the morbidity of radical radiotherapy for medically inoperable NSCLC. SEARCH STRATEGY: Randomised trials were sought by electronic searching the Cochrane Clinical Trials Register and both randomised and non-randomised trials sought by searching Medline and Excerpta Medica (Embase). Further studies were identified from references cited in those papers already identified by electronic searching. SELECTION CRITERIA: Studies of patients of any age with stage I/II NSCLC receiving radiotherapy at a dose greater than 40Gy in 20 fractions over four weeks or its radiobiological equivalent. DATA COLLECTION AND ANALYSIS: Two randomised and thirty-five non-randomised studies were identified. One randomised and nine non-randomised studies did not meet the selection criteria and were not included in the review. MAIN RESULTS: In the randomised trial comparing two radiotherapy schedules, two-year survival was superior following continuous hyperfractionated accelerated radiotherapy (CHART; 37%) compared to 60Gy in 30 fractions over six weeks (24%). There were 26 non-randomised retrospective studies including an estimated 2003 patients, in which overall survival results varied between 33-72% at two years, 17-55% at three years and 0-42% at five years. The proportion of deaths not due to cancer was 11-43%. Cancer-specific survival was between 54-93% at two years, 22-56% at three years and 13-39% at five years. Complete response rates were 33-61% and local failure rates between 6 70%. Distant metastases developed in approximately 25% of patients. Better response rates and survival were seen in those with smaller tumours and in those receiving higher doses though the reasons for prescribing higher doses were not clearly stated. Worse outcome was seen in those with prior weight loss or poor performance status. Assessment of treatment-related morbidity and effects on quality of life and symptom control were inconclusive because of the lack of prospective evaluation and paucity of data. REVIEWER'S CONCLUSIONS: There were no randomised trials that compared a policy of immediate radical radiotherapy with palliative radiotherapy given when patients develop symptoms. In the absence of such trials, radical radiotherapy appears to result in a better survival than might be expected had treatment not been given. A substantial, though variable, proportion of patients died during follow-up from causes other than cancer. The optimal radiation dose and treatment technique (particularly with respect to mediastinal irradiation) remain uncertain. PMID- 11406052 TI - Inhaled short-acting beta2-agonists versus ipratropium for acute exacerbations of chronic obstructive pulmonary disease. AB - BACKGROUND: Inhaled short acting beta2 adrenergic agonists and ipratropium bromide are both used in the treatment of acute exacerbations of chronic obstructive pulmonary disease. OBJECTIVES: In patients with acute exacerbations of COPD to: 1. To assess the efficacy of short-acting beta-2 agonists against placebo; 2. Compare the efficacy of short-acting beta-2 agonists and ipratropium. SEARCH STRATEGY: A comprehensive search of the literature was carried out of EMBASE, MEDLINE, CINAHL and the Cochrane COPD trials register was carried out using the terms: bronchodilator* OR albuterol OR metaproterenol OR terbutaline OR isoetharine OR pirbuterol OR salbutamol OR beta-2 agonist. SELECTION CRITERIA: All trials that appeared to be relevant were assessed by two reviewers who independently selected trials for inclusion. Differences were resolved by consensus. DATA COLLECTION AND ANALYSIS: All trials that appeared to be relevant were assessed by two reviewers who independently selected trials for inclusion. Differences were resolved by consensus. References listed in each included trial were searched for additional trial reports. Trials were combined using Review Manager using a fixed effects model. The size of the treatment effects were tested for heterogeneity. MAIN RESULTS: We identified no placebo-controlled comparisons of beta-2 agonists. Three studies permitted comparison of ipratropium to an inhaled beta-2 agonist. These studies included a total of 103 patients. The beta2-agonists used were: fenoterol and metaproterenol. One study was a parallel group trial of regular therapy for seven days. The other two were cross over studies of single dose treatments, with efficacy measured 90 min post dose. There was no washout period between treatments. Both treatments produced an improvement in forced expiratory volume (FEV1) after 90 min in the range 150-250 ml. The was no difference between treatments, mean difference in FEV1 10 ml; 95% CI -220, 230 ml. In one small crossover study (n=10) there was a significant improvement in arterial PaO2 after 30 minutes with ipratropium (+5.8 mm Hg +/- 3.0 (SEM)) compared to metaproterenol (-6.2 +/- 1.2 mm Hg), but this was not significant at 90 min. There were no data concerning respiratory symptoms. The crossover studies showed no evidence of an additive effect of the two treatments, although they were not designed specifically to test this. REVIEWER'S CONCLUSIONS: There are few controlled trial data concerning the use of inhaled beta2-agonist agents in acute exacerbations of COPD and none that have compared these agents directly with placebo. None of the studies used the more modern beta2-agonists used most widely in this setting (salbutamol and terbutaline). Beta2-agonists and ipratropium both produce small improvements in FEV1, but beta2-agonists may worsen PaO2 for a period. We could not draw conclusions concerning possible additive effects. PMID- 11406053 TI - Gold as an oral corticosteroid sparing agent in stable asthma. AB - BACKGROUND: Patients with chronic severe asthma are often dependent on the long term prescription of oral corticosteroids. The use of steroids is associated with serious side effects. Physicians treating such patients continue to search for alternative therapies that reduce the need for chronic dosing with oral steroids. Gold compounds are immunosuppressive agents and have benefits in the treatment of a number of inflammatory disorders. They have therefore been identified as an potentially useful agents in the treatment of chronic severe asthma both in terms of possible efficacy and as steroid sparing agents. OBJECTIVES: The objective of this review was to assess the effects of adding gold to oral steroids in the treatment of chronic steroid dependent asthmatics. SEARCH STRATEGY: The Cochrane Airways Group trials register and reference lists of identified articles were searched. SELECTION CRITERIA: Randomised trials looking at the addition of gold compared to placebo in adult steroid dependent asthmatics. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extraction was carried out by two reviewers independently. Study authors were contacted for missing information. MAIN RESULTS: Three trials fulfilled the criteria for inclusion in the review and a total of 376 patients were recruited into these studies. Data from 311 patients could be analysed. There was a small but significant treatment effect for gold in terms of steroid dose reduction (Peto Odds Ratio 0.51, 95% confidence intervals 0.31,0.83). No meta-analysis could be done for measures of lung function although overall there were few changes suggesting a positive benefit for gold. There were trends suggestive of adverse effects but no significant changes for gold treated patients with respect to proteinuria (Peto Odds Ratio 1.4, 95% confidence intervals 0.6, 3.3) dermatitis/eczema Peto Odds Ratio 2.1, 95% confidence intervals 0.9, 4.7). REVIEWER'S CONCLUSIONS: The changes seen in these trials are small and probably of limited clinical significance. Given the side effects of gold and necessity for monitoring the use of gold as a steroid sparing agent in asthma cannot be recommended. PMID- 11406054 TI - Troleandomycin as an oral corticosteroid steroid sparing agent in stable asthma. AB - BACKGROUND: Patients with chronic severe asthma are often dependent on the long term prescription of oral corticosteroids. The use of steroids is associated with serious side effects. Physicians treating such patients continue to search for alternative therapies that reduce the need for chronic dosing with oral steroids. troleandomycin is a compound that is established as an effective antibiotic but may also have non antibacterial actions that may be useful in the treatment of asthma. OBJECTIVES: The objective of this review was to assess the effects of adding troleandomycin to oral steroids in the treatment of chronic steroid dependent asthmatics. SEARCH STRATEGY: The Cochrane Airways Group trials register and reference lists of identified articles were searched. SELECTION CRITERIA: Randomised trials looking at the addition of troleandomycin compared to placebo in adult steroid dependent asthmatics. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extraction was carried out by two reviewers independently. Study authors were contacted for missing information. MAIN RESULTS: Three trials fulfilled the criteria for inclusion in the review and a total of 112 patients were recruited into these studies. Data from 90 patients were analysed. There was no treatment effect for troleandomycin in terms of steroid dose reduction (SMD -0.29, 95% CI -0.75, 0.17). For measures of lung function a meta-analysis of data derived from two of the included studies showed no benefits for added troleandomycin (SMD 0.06 95% CI -0.8, 0.9). REVIEWER'S CONCLUSIONS: There is insufficient evidence to support the use of troleandomycin in the treatment of steroid dependent asthma. PMID- 11406055 TI - Intravenous beta2-agonists for acute asthma in the emergency department. AB - BACKGROUND: Inhaled beta-agonist therapy is central to the management of acute asthma. The use of intravenous beta-agonist agents may also be beneficial in this setting. OBJECTIVES: To determine the benefit of intravenous (IV) beta2-agonists for severe acute asthma treated in the emergency department. SEARCH STRATEGY: Randomised controlled trials (RCT) were identified using the Cochrane Airways Group Register which is a compilation of systematic searches of MEDLINE, EMBASE, CINAHL, and CENTRAL as well as hand searching of 20 respiratory journals. Bibliographies from included studies and known reviews were also searched. Primary authors and content experts were contacted to identify eligible studies. SELECTION CRITERIA: Only RCTs were considered for inclusion. Studies were included if patients presented to the emergency department with acute asthma and were treated with IV selective or nonselective beta2-agonists versus placebo, inhaled beta2-agonists, or other standard of care. Pulmonary function, vital signs, arterial gasses, adverse effects, and/or clinical success could be reported as outcome measures. Two reviewers independently selected potentially relevant articles and selected articles for inclusion. Methodological quality was independently assessed using two scoring systems and two reviewers. DATA COLLECTION AND ANALYSIS: Data were extracted independently by two reviewers, and confirmed with corresponding authors. Missing data were obtained from authors or calculated from data present in the papers. Trials were combined using a random effects model for odds ratios (OR) or weighted mean differences (WMD) and reported with 95% confidence intervals (95% CI). MAIN RESULTS: From 746 identified references, 55 potentially relevant articles were identified and 15 were included. The trials included 584 patients. Overall, selective IV beta2 agonist use conferred no advantage over the comparator regimes. For example, it was associated with a lower PEFR after 60 mins compared to inhaled beta2-agonist, although the difference was not statistically significant (-24.7 l/min; 95%CI 2.9, -52.3). There was no difference in heart rate (4.5 bpm; 95% CI -4.9, 14.0). In the well performed blinded studies there was no difference in autonomic side effects between treatments (Odds Ratio 2.2 (95%CI 0.9, 5.7). REVIEWER'S CONCLUSIONS: There is no evidence to support the use of IV beta2-agonists in patients with severe acute asthma. These drugs should be given by inhalation. No subgroups were identified in which the IV route should be considered. PMID- 11406056 TI - Cardioselective beta-blocker use in patients with reversible airway disease. AB - BACKGROUND: Beta-blocker therapy has mortality benefit in patients with hypertension, heart failure and coronary artery disease, as well as during the perioperative period. These drugs have traditionally been considered contraindicated in patients with reversible airway disease. OBJECTIVES: To assess the effect of cardioselective beta-blockers on respiratory function of patients with reversible airway disease. Reversible airway disease was defined as asthma or chronic obstructive pulmonary disease with a reversible obstructive component. SEARCH STRATEGY: A comprehensive search of EMBASE, MEDLINE and CINAHL was performed using the Cochrane Airways Group registry to identify randomized blinded placebo-controlled trials from 1966 to February, 2000. The search was completed using the terms: asthma*, bronchial hyperreactivity*, respiratory sounds*, wheez*, obstructive lung disease* or obstructive airway disease*, and adrenergic antagonist*, sympatholytic* or adrenergic receptor block*. We did not exclude trials on the basis of language. SELECTION CRITERIA: Randomized, blinded, placebo-controlled trials of single dose or longer duration that studied the effects of cardioselective beta-blockers on the forced expiratory volume in 1 second (FEV1), symptoms and use of short-acting inhaled beta-agonists, in patients with reversible airway disease. Reversible airway disease was documented by response to methacholine challenge, by an increase in FEV1 of at least 15% to beta-agonist administration, or the presence of asthma as defined by the American Thoracic Society. DATA COLLECTION AND ANALYSIS: Two independent reviewers extracted data from the selected articles, reconciling differences by consensus. Cardioselective beta-blockers were divided into 2 groups, those with or without intrinsic sympathomimetic activity (ISA). Two interventions studied were the administration of beta-blocker, given either as a single dose or for longer duration, and the use of beta-agonist given after the study drug. MAIN RESULTS: Nineteen studies for single-dose treatment and 10 for treatment of longer duration met selection criteria. The patients had mild-moderate airways obstruction. For cardioselective beta-blockers taken as a group, administration of a single dose was associated with a 7.98% (CI, 6.19 to 9.77%) reduction in FEV1, but with a 13.16% (CI, 10.76 to 15.56%) increase in beta-agonist response, as compared to placebo. There was no increase in symptoms. After treatment lasting a few days to a few weeks, there was no decrement in FEV1 compared to placebo and no increase in symptoms or inhaler use. Regular use of cardioselective beta-blockers without ISA produced a 13.13% (CI, 5.97 to 20.30) increase in beta-agonist response compared to placebo, a response not seen with beta-blockers containing ISA (-0.60% [CI, -11.7 to +10.5%]). REVIEWER'S CONCLUSIONS: Cardioselective beta-blockers, given to patients with mild-moderate reversible airway disease, do not produce clinically significant adverse respiratory effects in the short term. It is not possible to comment on their effects in patient with more severe or less reversible disease, or on their effect on the frequency or severity of acute exacerbations. Given their demonstrated benefit in conditions such as heart failure, coronary artery disease and hypertension, cardioselective beta-blockers should not be withheld from patients with mild-moderate reversible airway disease. PMID- 11406057 TI - Cyclosporin as an oral corticosteroid sparing agent in stable asthma. AB - BACKGROUND: Patients with chronic severe asthma are often dependent on the long term prescription of oral corticosteroids. The use of steroids is associated with serious side effects. Physicians treating such patients continue to search for alternative therapies that reduce the need for chronic dosing with oral steroids. Cyclosporin is an immunosuppressive agent and has benefits in the treatment of a number of inflammatory disorders. It has therefore been identified as an potentially useful agent in the treatment of chronic severe asthma both in terms of possible efficacy and as a steroid sparing agent. OBJECTIVES: The objective of this review was to assess the effects of adding cyclosporin to oral steroids in the treatment of chronic steroid dependent asthmatics. SEARCH STRATEGY: The Cochrane Airways Group trials register and reference lists of identified articles were searched. SELECTION CRITERIA: Randomised trials looking at the addition of cyclosporin compared to placebo in adult steroid dependent asthmatics. DATA COLLECTION AND ANALYSIS: Trial quality was assessed and data extraction was carried out by two reviewers independently. Study authors were contacted for missing information. MAIN RESULTS: Three trials fulfilled the criteria for inclusion in the review and a total of 106 patients were recruited into these studies. Data from 98 patients could be analysed. There was a small but significant treatment effect for cyclosporin in terms of steroid dose reduction (SMD -0.5, 95% CI -1.0, -0.04). No meta-analyses could be performed for measures of lung function although one study showed small, but significant improvements in lung spirometry. REVIEWER'S CONCLUSIONS: The changes with cyclosporin are small and of questionable clinical significance. Given the side effects of cyclosporin, the evidence available does not recommend routine use of this drug in the treatment of oral corticosteroid dependent asthma. PMID- 11406058 TI - Inhaled hyperosmolar agents for bronchiectasis. AB - BACKGROUND: Mucus retention in the lungs is a prominent feature of bronchiectasis. The stagnant mucus becomes chronically colonised with bacteria, which elicit a host neutrophilic response. This fails to eliminate the bacteria, and the large concentration of host-derived protease may contribute to the airway damage. The sensation of retained mucus is itself a cause of suffering, and the failure to maintain airway sterility probably contributes to the frequent respiratory infections experienced by many patients. Hypertonic saline inhalation is known to accelerate tracheobronchial clearance in many conditions, probably by inducing a liquid flux into the airway surface, which alters mucus rheology in a way favourable to mucociliary clearance. Inhaled dry powder mannitol has a similar effect. Such agents are an attractive approach to the problem of mucostasis, and deserve further clinical evaluation. OBJECTIVES: To determine whether inhaled hyperosmolar substances are efficacious in the treatment of bronchiectasis SEARCH STRATEGY: MEDLINE and Cochrane databases were searched, and leaders in the field contacted. SELECTION CRITERIA: Any trial using hyperosmolar inhalation in patients with bronchiectasis not caused by cystic fibrosis. DATA COLLECTION AND ANALYSIS: One reference were identified by the searches conducted. MAIN RESULTS: Only one trial was identified, a crossover study of 11 patients with bronchiectasis. The outcome measure was tracheobronchial clearance of a particulate radioaerosol after inhalation of dry mannitol on a single occasion, with appropriate controls. Airway clearance doubled in the central and intermediate regions of the lung, but not in the peripheral region, after mannitol administration. No side effects were observed, but two patients were premedicated with nedocromil to prevent bronchospasm. REVIEWER'S CONCLUSIONS: Dry powder mannitol has been shown to improve tracheobronchial clearance in bronchiectasis, as well as cystic fibrosis, asthmatics, and normal subjects. It is not yet available for clinical use. Hypertonic saline has not been specifically tested in bronchiectasis, but improve clearance in these other conditions and in chronic bronchitis. Longer term randomised controlled studies of mannitol and hypertonic saline with clinical endpoints are now needed. PMID- 11406059 TI - Transcutaneous electrical nerve stimulation (TENS) for chronic low back pain. AB - BACKGROUND: Low back pain (LBP) affects a large proportion of the population. Transcutaneous electrical nerve stimulation (TENS) was introduced more than 30 years ago as an alternative therapy to pharmacological treatments for chronic pain. However, despite its widespread use, the effectiveness of TENS is still controversial. OBJECTIVES: The aim of this systematic review was to determine the efficacy of TENS in the treatment of chronic LBP. SEARCH STRATEGY: We searched MEDLINE, EMBASE, PEDro and the Cochrane Controlled Trials Register up to June 1, 2000. SELECTION CRITERIA: Only randomized controlled clinical trials of TENS for the treatment of patients with a clinical diagnosis of chronic LBP were included. Abstracts were excluded unless further data could be obtained from the authors. DATA COLLECTION AND ANALYSIS: Two reviewers independently selected trials and extracted data using predetermined forms. Heterogeneity was tested with Cochran's Q test. A fixed effects model was used throughout for continuous variables, except where heterogeneity existed, in which case, a random effects model was used. Results are presented as weighted mean differences (WMD) with 95% confidence intervals (95% CI), where the difference between the treated and control groups was weighted by the inverse of the variance. Standardized mean differences (SMD) were calculated by dividing the difference between the treated and control by the baseline variance. SMD were used when different scales were used to measure the same concept. Dichotomous outcomes were analyzed with odds ratios. MAIN RESULTS: Five trials were included, with 170 subjects randomized to the placebo group receiving sham-TENS and 251 subjects receiving active TENS (153 for conventional mode, 98 for acupuncture-like TENS). The schedule of treatments varied greatly between studies ranging from one treatment/day for two consecutive days, to three treatments/day for four weeks. There were no statistically significant differences between the active TENS group when compared to the placebo TENS group for any outcome measures. Subgroup analysis performed on TENS application and methodological quality did not demonstrate a significant statistical difference. Remaining pre-planned subgroup analysis was not conducted due to the small number of included trials and the variety of outcome measures reported. REVIEWER'S CONCLUSIONS: The results of the meta-analysis present no evidence to support the use of TENS in the treatment of chronic low back pain. Clinicians and researchers should consistently report the characteristics of the TENS device and the application techniques used. New trials on TENS should make use of standardized outcome measures. This meta-analysis lacked data on how TENS effectiveness is affected by four important factors: type of applications, site of application, treatment duration of TENS, optimal frequencies and intensities. PMID- 11406060 TI - Antidepressants using active placebos. AB - BACKGROUND: Although there is a consensus that antidepressants are effective in depression, placebo effects are also thought to be substantial. Side effects of antidepressants may reveal the identity of medication to participants or investigators and thus may bias the results of conventional trials using inert placebos. Using an 'active' placebo which mimics some of the side effects of antidepressants may help to counteract this potential bias. OBJECTIVES: To investigate the efficacy of antidepressants when compared with 'active' placebos. SEARCH STRATEGY: The Cochrane Collaboration Depression, Anxiety and Neurosis review groups's search strategy was used to search MEDLINE (1966-2000), PsychLIT (1980-2000) and EMBASE (1974-2000) and this was last done in July 2000. Reference lists from relevant articles and textbooks were searched and 12 specialist journals were handsearched up to 1996. SELECTION CRITERIA: Randomised and quasi randomised controlled trials comparing antidepressants with active placebos in people with depression. DATA COLLECTION AND ANALYSIS: Since many different outcome measures were used a standard measure of effect was calculated for each trial. A subgroup analysis of inpatient and outpatient trials was conducted. Two reviewers independently assessed whether each trial met inclusion criteria. MAIN RESULTS: Nine studies involving 751 participants were included. Two of them produced effect sizes which showed a consistent and statistically significant difference in favour of the active drug. Combining all studies produced a pooled estimate of effect of 0.39 standard deviations (confidence interval, 0.24 to 0.54) in favour of the antidepressant measured by improvement in mood. There was high heterogeneity due to one strongly positive trial. Sensitivity analysis omitting this trial reduced the pooled effect to 0.17 (0.00 to 0.34). The pooled effect for inpatient and outpatient trials was highly sensitive to decisions about which combination of data was included but inpatient trials produced the lowest effects. REVIEWER'S CONCLUSIONS: The more conservative estimates from the present analysis found that differences between antidepressants and active placebos were small. This suggests that unblinding effects may inflate the efficacy of antidepressants in trials using inert placebos. Further research into unblinding is warranted. PMID- 11406061 TI - Lithium for maintenance treatment of mood disorders. AB - BACKGROUND: Mood disorders are common, disabling and tend to be recurrent. They carry a high risk of suicide. Maintenance treatment, aimed at the prevention of relapse, is therefore of vital importance. Lithium has been used for some years as the mainstay of maintenance treatment in bipolar affective disorder, and to a lesser extent in unipolar disorder. However, the efficacy and effectiveness of prophylactic lithium therapy has been disputed. Low suicide rates in lithium treated patients have led to claims that lithium has a specific anti-suicidal effect. If so, this is of considerable importance as treatments for mental disorders in general have not been shown convincingly to be effective in suicide prevention. OBJECTIVES: 1. To investigate the efficacy of lithium treatment in the prevention of relapse in recurrent mood disorders. 2. To examine the effect of lithium treatment on consumers' general health and social functioning, its acceptability to consumers, and the side-effects of treatment. 3. To investigate the hypothesis that lithium has a specific effect in reducing the incidence of suicide and deliberate self-harm in persons with mood disorders. SEARCH STRATEGY: The Cochrane Collaboration Depression, Anxiety and Neurosis Controlled Trials Register (CCDANCTR) and The Cochrane Controlled Clinical Trials Register (CCTR) were searched. Reference lists of relevant papers and major text books of mood disorder were examined. Authors, other experts in the field and pharmaceutical companies were contacted for knowledge of suitable trials, published or unpublished. Specialist journals concerning lithium were hand searched. SELECTION CRITERIA: Randomised controlled trials comparing lithium with placebo, where the stated intent of treatment was maintenance or prophylaxis. Participants were males and females of all ages with diagnoses of mood disorder. Discontinuation studies (in which all participants had been stable on lithium for some time before being randomised to either continued lithium treatment or placebo substitution) were excluded. DATA COLLECTION AND ANALYSIS: Data were extracted from the original reports independently by two reviewers. The main outcomes studied were related to the objectives stated above. Data were analysed for all diagnoses of mood disorder and for bipolar and unipolar disorder separately. Data were analysed using Review Manager version 4.0. MAIN RESULTS: Nine studies were included in the review, reporting on 825 participants randomly allocated to lithium or placebo. Lithium was found to be more effective than placebo in preventing relapse in mood disorder overall, and in bipolar disorder. The most consistent effect was found in bipolar disorder (random effects OR 0.29; 95% CI 0.09 to 0.93 ). In unipolar disorder, the direction of effect was in favour of lithium, but the result (when heterogeneity between studies was allowed for) did not reach statistical significance. Considerable heterogeneity was found between studies in all groups of patients. The direction of effect was the same in all studies; no study found a negative effect for lithium. Heterogeneity may have been due to differences in selection of participants, and to differing exposures to lithium in the pre-study phase resulting in variable influence of a discontinuation effect. There was little reported data on overall health and social functioning of participants under the different treatment conditions, or on the participants' own views of their treatment. Descriptive analysis showed that assessments of general health and social functioning generally favoured lithium. Small absolute numbers of deaths and suicides, and the absence of data on non-fatal suicidal behaviours, made it impossible to draw meaningful conclusions about the place of lithium therapy in suicide prevention. REVIEWER'S CONCLUSIONS: This systematic review indicates that lithium is an efficacious maintenance treatment for bipolar disorder. In unipolar disorder the evidence of efficacy is less robust. This review does not cover the relative efficacy of lithium compared with other maintenance treatments, which is at present unclear. There is no definitive evidence from this review as to whether or not lithium has an anti-suicidal effect. Systematic reviews and large scale randomised studies comparing lithium with other maintenance treatments (e.g. anti-convulsants, antidepressants) are necessary. Outcomes relating to death and suicidal behaviour should be included in all future maintenance studies of mood disorder. PMID- 11406062 TI - Family and parenting interventions in children and adolescents with conduct disorder and delinquency aged 10-17. AB - BACKGROUND: Conduct disorder and delinquency are significant problems for children and adolescents and their families, with the potential to consume much of the resources of the health, social care and juvenile justice systems. A number of family and parenting interventions have been recommended and are used for these conditions. The aim of this review was to determine if these interventions are effective in the management of conduct disorder and delinquency in children and adolescents, aged 10-17. OBJECTIVES: To determine if family and parenting interventions improve the child/adolescent's behaviour; parenting and parental mental health; family functioning and relations; and have an effect on the long term psychosocial outcomes for the child/adolescent. SEARCH STRATEGY: Randomised controlled trials were identified through searching the Cochrane Controlled Trial Register (CCTR), databases (MEDLINE, EMBASE, PsycINFO, CINAHL, Sociofile, ERIC, Healthstar), reference lists of articles and contact with authors. SELECTION CRITERIA: Randomised controlled trials with a major focus on parenting and/or family functioning were eligible for inclusion in the review. Trials needed to include at least one objective outcome measure (e.g. arrest rates) or have used a measure that had been published in peer review publications and validated for the relevant purpose. Studies were required to have a control group, which could be a no intervention group, a wait list group or a usual intervention group (e.g. probation). Trials in children and adolescents aged 10 to 17 years with conduct disorder and/or delinquency and their families were considered. Conduct disorder was defined by a standardised psychological assessment (for example, using a child behaviour checklist), or a psychiatric diagnosis. Delinquency was defined by a referral from a juvenile justice or another legal system for a child/adolescent who has committed a serious crime e.g assault and/or offended on at least two occasions. DATA COLLECTION AND ANALYSIS: Two reviewers independently reviewed all eligible studies for inclusion, assessed study quality (allocation concealment, blinding, follow up, clinically important outcomes) and extracted data. Heterogeneity was assessed using the Chi squared test of heterogeneity along with visual inspection of the data. A significance level less than 0.1 was interpreted as evidence of statistically significant heterogeneity. For data where heterogeneity was found the reviewers looked for an explanation. If studies with heterogeneous results were thought to be comparable the statistical synthesis of the results was done using a random effects model. This model takes into account within-study sampling error and between-studies variation in the assessment of uncertainty and will give wider confidence intervals to the effect size and hence a more conservative result. Sensitivity analysis was performed to explore the effects of the varying quality of the studies included on the results. MAIN RESULTS: Of the nine hundred and seventy titles initially identified through the search strategy, eight trials met the inclusion criteria. A total of 749 children and their families were randomised to receive a family and parenting intervention or to be in a control group. In seven of these studies the participants were juvenile delinquents and their families and in only one the participants were children/adolescents with conduct disorder who had not yet had contact with the juvenile justice system. At follow up, family and parenting interventions significantly reduced the time spent by juvenile delinquents in institutions (WMD 51.34 days, 95%CI 72.52 to 30.16). There was also a significant reduction in the risk of a juvenile delinquent being re arrested (RR 0.66, 95%CI 0.44 to 0.98) and in their rate of subsequent arrests at 1-3 years (SMD -0.56, 95% CI -1.100 to - 0.03). For both of these outcomes there was substantial heterogeneity in the results suggesting a need for caution in interpretation. At present there is insufficient evidence that family and parenting interventions reduce the risk of being incarcerated (RR=0.50, 95% CI 0.20 to 1.21). No significant difference was found for psychosocial outcomes such as family functioning, and child/adolescent behaviour. REVIEWER'S CONCLUSIONS: The evidence suggests that family and parenting interventions for juvenile delinquents and their families have beneficial effects on reducing time spent in institutions. This has an obvious benefit to the participant and their family and may result in a cost saving for society. These interventions may also reduce rates of subsequent arrest but at present these results need to be interpreted with caution due to the heterogeneity of the results. PMID- 11406063 TI - Continuing education meetings and workshops: effects on professional practice and health care outcomes. AB - BACKGROUND: Educational meetings and printed educational materials are the two most common types of continuing education for health professionals. An important aim of continuing education is to improve professional practice so that patients can receive improved health care. OBJECTIVES: To assess the effects of educational meetings on professional practice and health care outcomes. SEARCH STRATEGY: We searched the Cochrane Effective Practice and Organisation of Care Group specialised register, MEDLINE (from 1966), the Research and Development Resource Base in Continuing Medical Education in January 1999 and reference lists of articles. SELECTION CRITERIA: Randomised trials or well designed quasi experimental studies examining the effect of continuing education meetings (including lectures, workshops, and courses) on the clinical practice of health professionals or health care outcomes. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied inclusion criteria, assessed the quality of each study, and extracted study data. We attempted to collect missing data from investigators. We conducted both qualitative and quantitative analyses. MAIN RESULTS: Thirty-two studies were included with a total of 36 comparisons. The studies involved from 13 to 411 health professionals (total N= 2995) and were judged to be of moderate or high quality, although methods were generally poorly reported. There was substantial variation in the complexity of the targeted behaviours, baseline compliance, the characteristics of the interventions and the results. The heterogeneity of the results was best explained by differences in the interventions. For 10 comparisons of interactive workshops, there were moderate or moderately large effects in six (all of which were statistically significant) and small effects in four (one of which was statistically significant). For interventions that combined workshops and didactic presentations, there were moderate or moderately large effects in 12 comparisons (eleven of which were statistically significant) and small effects in seven comparisons (one of which was statistically significant). In seven comparisons of didactic presentations, there were no statistically significant effects, with the exception of one out of four outcome measures in one study. REVIEWER'S CONCLUSIONS: Interactive workshops can result in moderately large changes in professional practice. Didactic sessions alone are unlikely to change professional practice. PMID- 11406064 TI - Immediate post-partum insertion of intrauterine devices. AB - OBJECTIVES: To assess the efficacy and feasibility of IUD insertion immediately after expulsion of the placenta. Our a priori hypothesis was that this practice is safe but associated with higher expulsion rates than interval IUD insertion. SEARCH STRATEGY: We used MEDLINE, Popline, EMBASE, and Cochrane Controlled Trials Register computer searches, supplemented by review articles and contact with investigators. SELECTION CRITERIA: We sought all randomized controlled trials that had at least one treatment arm that involved immediate post-partum (within ten minutes of placental expulsion) insertion of an IUD. Comparisons could include different IUDs, different insertion techniques, immediate vs. delayed post-partum insertion, or immediate vs. interval insertion (unrelated to pregnancy). Studies could include either vaginal or cesarean deliveries. DATA COLLECTION AND ANALYSIS: We evaluated the methodological quality of each report and sought to identify duplicate reporting of data from multicenter trials. We abstracted data onto data collection forms. Principal outcome measures included pregnancy, expulsion, and continuation rates. Because the trials did not have uniform interventions, we were unable to aggregate them in a meta-analysis. MAIN RESULTS: We found no randomized controlled trials that directly compared immediate post-partum insertion with either delayed post-partum or interval insertion. Modifications of existing devices, such as adding absorbable sutures or additional appendages, did not appear beneficial. Most studies showed no important differences between insertions done by hand or by instruments. Lippes Loops and Progestasert devices did not perform as well as did copper devices. REVIEWER'S CONCLUSIONS: Immediate post-partum insertion of IUDs appeared safe and effective, though direct comparisons with other insertion times were lacking. Advantages of immediate post-partum insertion include high motivation, assurance that the woman is not pregnant, and convenience. However, expulsion rates appear to be higher than with interval insertion. The popularity of immediate post partum IUD insertion in countries as diverse as China, Mexico, and Egypt support the feasibility of this approach. Early follow-up may be important in identifying spontaneous IUD expulsions. PMID- 11406065 TI - Cromolyn sodium for the prevention of chronic lung disease in preterm infants. AB - BACKGROUND: Chronic lung disease (CLD) frequently occurs in preterm infants (< 37 weeks gestational age) and has a multifactorial etiology including inflammation. Cromolyn sodium is a mast cell stabiliser that inhibits neutrophil activation and neutrophil chemotaxis. Therefore it is possible that cromolyn sodium might have a role in the prevention of CLD. OBJECTIVES: QUESTION: in preterm infants, does the prophylactic administration of cromolyn sodium reduce the incidence of CLD, mortality or the combined outcome of mortality or CLD at 28 days of life without undue side effects? SEARCH STRATEGY: The search strategy used to identify studies was according to the guidelines of the Cochrane Neonatal Review Group. Searches were made of MEDLINE, EMBASE, CINAHL up to and including December 2000, the Cochrane Library 2000 Issue 4, personal files and reference lists of identified trials. The following terms were used: chronic lung disease, cromolyn sodium or cromoglycate. LIMITS: newborn, infant; human, clinical trial or controlled clinical trial or meta analysis or randomised controlled trial. No language restrictions were applied. SELECTION CRITERIA: Randomised or quasi-randomised controlled clinical trials involving preterm infants. Initiation of cromolyn sodium administration during the first two weeks of life. The intervention had to include administration of cromolyn sodium by nebulizer or metered dose inhaler with or without spacer device, versus placebo or no intervention. Eligible studies had to include at least one of the following outcomes: overall mortality, CLD at 28 days, CLD at 36 weeks corrected gestational age, or the combined outcome mortality or CLD at 28 days. Secondary outcomes included number of days on oxygen, number of days on mechanical ventilation, patent ductus arteriosus (PDA), air leaks [pulmonary interstitial emphysema (PIE), pneumothorax], any grade of intraventricular haemorrhage (IVH), necrotizing enterocolitis (NEC), sepsis and adverse effects due to cromolyn sodium. DATA COLLECTION AND ANALYSIS: We used the standard method for the Cochrane Collaboration as described in the Cochrane Collaboration handbook. Both investigators extracted and assessed all data for each study. Any disagreement was resolved by discussion. Relative risk (RR) and risk difference (RD) with 95% confidence intervals (CI) are reported for dichotomous outcomes and weighted mean difference (WMD) for continuous data. Number needed to treat was not calculated as no outcome showed a statistically significant RD. A fixed effect model was used for meta-analysis. MAIN RESULTS: Two eligible studies were identified with small numbers of infants enrolled. Prophylaxis with cromolyn sodium did not result in a statistically significant effect on the combined outcome, death or CLD at 28 days [RR 1.05 (95% CI 0.73, 1.52); RD 0.03 (95% CI -0.20, 0.27)], CLD at 28 days [RR 0.93 (95% CI 0.53, 1.64; RD -0.03 (95% CI -0.27, 0.20)], CLD at 36 weeks corrected gestational age [RR 1.25 (95% CI 0.43, 3.63); RD 0.08 (95% CI -0.29, 0.44)], CLD in survivors at 28 days [RR 0.97 (95% CI 0.58, 1.63); RD -0.02 (95% CI -0.29, 0.26)] or CLD in survivors at 36 weeks corrected gestational age [RR 1.04 (95% CI 0.38, 2.87); RD 0.02 (95% CI -0.40, 0.43)]. Prophylaxis with cromolyn sodium did not show a statistically significant difference in overall mortality [RR 1.31 (95% CI 0.52, 3.29); RD 0.06 (95% CI -0.13, 0.26)]. There were no statistically significant differences in the incidence of air leaks, NEC, IVH, sepsis, days on mechanical ventilation or PDA. No side effects were noted. REVIEWER'S CONCLUSIONS: There is currently no evidence from randomized trials that cromolyn sodium has a role in the prevention of CLD. Cromolyn sodium cannot be recommended for the prevention of CLD in preterm infants. Additional clinical trials do not appear to be justified using the protocols for drug administration used to date unless a more efficient type of delivery device than the jet nebulizer is employed. PMID- 11406066 TI - Tracheal suctioning without disconnection in intubated ventilated neonates. AB - BACKGROUND: Assisted mechanical ventilation is the mainstay of management of a variety of conditions affecting the neonate. However there are a number of potential hazards associated with this life saving intervention. New suctioning techniques have been introduced into clinical practice which aim to prevent or reduce these untoward effects. OBJECTIVES: To assess the effects of endotracheal suctioning without disconnection in intubated ventilated neonates. SEARCH STRATEGY: The review has drawn on the search strategy for the Cochrane Neonatal Review Group. A comprehensive search of MEDLINE, Cochrane Library, MEDLINE and CINAHL databases was undertaken by the reviewers. SELECTION CRITERIA: All trials utilizing random or quasi-random patient allocation in which suctioning with or without disconnection from the ventilator is compared in the neonatal population. DATA COLLECTION AND ANALYSIS: Standard methods of the Cochrane Neonatal Group were used. Each author reviewed trials for eligibility and quality and extracted data separately, then compared and resolved differences. Analysis was performed using the fixed effects model and outcomes were reported using relative risk for categorical data and weighted mean difference for outcomes measured on a continuous scale. MAIN RESULTS: Two trials (22 infants) were included in this review. The trials employed a cross-over design in which suctioning with or without disconnection was compared. Suctioning without disconnection resulted in a reduction in episodes of hypoxia (RR 0.30, 95% CI 0.11, 0.80) and a smaller percentage decrease in the TcPO2 (WMD 18.5%, 95% CI 8.11, 28.89). There were also fewer infants who experienced episodes where TcPO2 decreased by > 10% (RR 0.36, 95% CI 0.17, 0.79). Suctioning without disconnection resulted in a smaller percentage decrease in heart rate (WMD 11.53%, 95% CI 3.64, 19.43) and a reduction in the number of infants experiencing a decrease in heart rate by > 10% (RR 0.56, 95% CI 0.32, 0.99). REVIEWER'S CONCLUSIONS: Based upon the results of this review, there is insufficient evidence to decide between endotracheal suctioning with or without disconnection. There is, however, evidence of some benefit from performing suctioning without disconnection for some specific short term outcomes. Further research should be undertaken to fully assess this practice with particular focus on extremely low birth weight infants and different modes of mechanical ventilation, and to address clinically important outcomes. PMID- 11406067 TI - Home versus in-patient treatment for deep vein thrombosis. AB - BACKGROUND: Deep vein thrombosis (DVT) affects 1-2/1,000 of the adult population per annum in western societies. It may be associated with pulmonary embolism (PE) which carries a 10% fatality rate. Sufferers may develop post-thrombotic syndrome with swelling of the leg, secondary varicose veins and ulceration. In the initial stages of treatment for DVT patients are traditionally admitted to hospital for intravenous treatment with unfractionated heparin (UH) for three to five days. The dose of UH required to provide a therapeutic level of anticoagulation is unpredictable, so the blood must be closely monitored. Fractionated, or low molecular weight heparin (LMWH), is given subcutaneously once daily and requires no monitoring, so can be given in hospital or at home. OBJECTIVES: To collate all randomised controlled trials (RCTs) comparing a home treatment regime (LMWH) with hospital treatment (LMWH or UH) for the initial phase of treatment for DVT, and to compare the safety, efficacy, patient acceptability and cost implications of home versus in-patient treatment. SEARCH STRATEGY: All published reports of home treatment were traced through MEDLINE, and EMBASE (up to and including December 2000) using the search strategy described by the Cochrane Peripheral Vascular Diseases Group. Additional searches included the Cochrane CCTR/CENTRAL database, handsearching non-listed journals, and personal communication with researchers. SELECTION CRITERIA: RCTs of home versus hospital treatment for DVT in which DVT was clinically confirmed and treated with either LMWH or UH. DATA COLLECTION AND ANALYSIS: One reviewer selected the material for inclusion (IGS): the other (AAM) reviewed the literature and selection of trials. Outcomes included PE, recurrent DVT, gangrene, heparin complications, and death. MAIN RESULTS: Only two major RCTs with comparable treatment arms were found. Both had fundamental problems including high exclusion rates, partial hospital treatment of many in the LMWH arms, and comparison of UH in hospital with LMWH at home. The trials showed that home treatment was no more liable to complications than hospital treatment. Initial results from a smaller RCT comparing LMWH treatment in both home and hospital arms came to the same conclusion. REVIEWER'S CONCLUSIONS: The limited evidence suggests that home management is cost effective, and likely to be preferred by patients. Further large trials comparing these treatments are unlikely to be held. Therefore, home treatment is likely to become the norm, and further research will be directed to resolving practical issues. PMID- 11406068 TI - Double bag or Y-set versus standard transfer systems for continuous ambulatory peritoneal dialysis in end-stage renal disease. AB - BACKGROUND: Peritonitis is the most frequent serious complication of continuous ambulatory peritoneal dialysis (CAPD). It has a major influence on the number of patients switching from CAPD to haemodialysis and has probably restricted the wider acceptance and uptake of CAPD as an alternative mode of dialysis. OBJECTIVES: This systematic review sought to determine if modifications of the transfer set (Y-set or double bag systems) used in CAPD exchanges are associated with a reduction in peritonitis and an improvement in other relevant outcomes. SEARCH STRATEGY: A broad search strategy was employed which attempted to identify all RCTs or quasi-RCTs relevant to the management of end-stage renal disease (ESRD). Five electronic databases were searched (Medline 1966-1999, EMBASE 1984 1999, CINAHL 1982-1996, BIOSIS 1985-1996 and the Cochrane Library), authors of included studies and relevant biomedical companies were contacted, reference lists of identified RCTs and relevant narrative reviews were screened and Kidney International 1980-1997 was hand searched. SELECTION CRITERIA: Randomised or quasi-randomised controlled trials comparing double bag, Y-set and standard CAPD exchange systems in patients with ESRD. DATA COLLECTION AND ANALYSIS: Data were abstracted by a single investigator onto a standard form and subsequently entered into Review Manager 4.0.4. Odds Ratio (OR) for dichotomous data and a (Weighted) Mean Difference (WMD) for continuous data were calculated with 95% confidence intervals (95% CI). MAIN RESULTS: Twelve eligible trials with a total of 991 randomised patients were identified. In trials comparing either the Y-set or double bag systems with the standard systems significantly fewer patients (OR 0.33, 95% CI 0.24 to 0.46) experienced peritonitis and the number of patient months on CAPD per episode of peritonitis were consistently greater. When the double bag systems were compared with the Y-set systems significantly fewer patients experienced peritonitis (OR 0.44, 95% CI 0.27 to 0.71) and the numbers of patient-months on CAPD/ episode of peritonitis were also greater. REVIEWER'S CONCLUSIONS: Double bag systems should be the preferred exchange systems in CAPD. PMID- 11406069 TI - Vocational rehabilitation for people with severe mental illness. AB - BACKGROUND: Unemployment rates are high amongst people with severe mental illness, yet surveys show that most want to work. Vocational rehabilitation services exist to help mentally ill people find work. Traditionally, these services have offered a period of preparation (Pre-vocational Training), before trying to place clients in competitive (i.e. open) employment. More recently, some services have begun placing clients in competitive employment immediately whilst providing on-the-job support (Supported Employment). It is unclear which approach is most effective. OBJECTIVES: To assess the effects of Pre-vocational Training and Supported Employment (for people with severe mental illness) against each other and against standard care (in hospital or community). In addition, to assess the effects of: (a) special varieties of Pre-vocational Training (Clubhouse model) and Supported Employment (Individual Placement and Support model); and (b) techniques for enhancing either approach, for example payment or psychological intervention. SEARCH STRATEGY: Searches were undertaken of CINAHL (1982-1998), The Cochrane Library (Issue 2, 1999), EMBASE (1980-1998), MEDLINE (1966-1998) and PsycLIT (1887-1998). Reference lists of eligible studies and reviews were inspected and researchers in the field were approached to identify unpublished studies. SELECTION CRITERIA: Randomised controlled trials of approaches to vocational rehabilitation for people with severe mental illness. DATA COLLECTION AND ANALYSIS: Included trials were reliably selected by a team of two raters. Data were extracted separately by two reviewers and cross-checked. Authors of trials were contacted for additional information. Relative risks (RR) and 95% confidence intervals (CI) of homogeneous dichotomous data were calculated. A random effects model was used for heterogeneous dichotomous data. Continuous data were presented in tables (there were insufficient continuous data for formal meta-analysis). A sensitivity analysis was performed, excluding poorer quality trials. MAIN RESULTS: Eighteen randomised controlled trials of reasonable quality were identified. The main finding was that on the primary outcome (number in competitive employment) Supported Employment was significantly more effective than Pre-vocational Training; for example, at 18 months 34% of people in Supported Employment were employed versus 12% in Pre-vocational Training (RR random effects (unemployment) 0.76 95% CI 0.64 to 0.89, NNT 4.5). Clients in Supported Employment also earned more and worked more hours per month than those in Pre-vocational Training. There was no evidence that Pre-vocational Training was more effective in helping clients to obtain competitive employment than standard community care. REVIEWER'S CONCLUSIONS: Supported employment is more effective than Pre-vocational Training in helping severely mentally ill people to obtain competitive employment. There is no clear evidence that Pre-vocational Training is effective. PMID- 11406070 TI - Haloperidol versus placebo for schizophrenia. AB - BACKGROUND: Haloperidol was developed in the late 1950s for use in the field of analgesia. Research subsequently demonstrated effects on hallucinations, delusions, aggressiveness, impulsiveness and states of excitement and led to the introduction of haloperidol as an antipsychotic. OBJECTIVES: To evaluate the clinical effects of haloperidol for the management of schizophrenia and other similar serious mental illnesses compared to placebo. SEARCH STRATEGY: Electronic searches of Biological Abstracts (1985-1998), CINAHL (1982-1998), The Cochrane Library (1998, Issue 4), The Cochrane Schizophrenia Group's Register (December 2000), EMBASE (1980-1998), MEDLINE (1966-1998), PsycLIT (1974-1998), and SCISEARCH (January 1974-December 1998) were undertaken. References of all identified studies were searched for further trial citations. Authors of trials and pharmaceutical companies were contacted for further information and archive material. SELECTION CRITERIA: All relevant randomised controlled trials comparing use of haloperidol (any dose) with placebo for those with schizophrenia or other similar serious, non-affective psychotic illnesses (however diagnosed). The main outcomes of interest were death, loss to follow up, clinical and social response, relapse and severity of adverse effects. DATA COLLECTION AND ANALYSIS: Reviewers evaluated data independently and analysed on an intention-to-treat basis, assuming that people who left the study early, or were lost to follow up, had no improvement. Where possible and appropriate, dichotomous data were analysed using relative risk (RR) and their 95% confidence intervals (CI) calculated. If appropriate, the number needed to treat (NNT) or number needed to harm (NNH) was estimated. For continuous data, weighted mean differences were calculated. Continuous data were excluded if loss to follow up was greater than 50%. All data were inspected for heterogeneity. MAIN RESULTS: Seventy-four trials were identified but only 20 included. More people allocated to haloperidol improved in the first six weeks of treatment than those given placebo (three trials, n=159, RR failing to produce a marked improvement 0.44 CI 0.3 to 0.6, NNT 3 CI 2 to 5). A further eight trials (n=313) also found a difference favouring haloperidol across the 6-24 week period (RR no marked global improvement 0.68 CI 0.6 to 0.8 NNT 3 CI 2.5 to 5) but this may be an overestimate of effect as small negative studies were not identified. About half of those entering studies failed to complete the short trials, although, at 0-6 weeks, 10 studies found a difference that favoured haloperidol (n=686, RR 0.82 CI 0.7 to 0.95, NNT 8 CI 5 to 17). Limited adverse effect data do, nevertheless, support the clinical impression that haloperidol is a potent cause of movement disorders, at least in the short term. Haloperidol promotes acute dystonia (three trials, n=135, RR 4.7 CI 1.7 to 44, NNH 5 CI 3 to 9 - not assuming those who left early from placebo suffered dystonis), akathisia (three trials, n=129, RR 6.5 CI 1.5 to 28, NNH 6 CI 4 to 14) and parkinsonism (four trials, n=165, RR 8.9 CI 2.6 to 31, NNH 3 CI 2 to 5). REVIEWER'S CONCLUSIONS: Haloperidol is a potent antipsychotic drug but with a high propensity to cause adverse effects. Given no choice of drug, use of haloperidol to counter the damaging and potentially dangerous consequences of untreated schizophrenia is justified. If a choice of drug is available, however, people with schizophrenia and clinicians may wish to start another antipsychotic with less likelihood of causing parkinsonism, akathisia and acute dystonias. For countries where haloperidol is not widely used, it should not be a control drug of choice for randomised trials of new antipsychotics. PMID- 11406071 TI - Extra-amniotic prostaglandin for induction of labour. AB - BACKGROUND: This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of extra-amniotic prostaglandin for third trimester cervical ripening or induction of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies of relevant papers. Date of last search: December 2000. SELECTION CRITERIA: (1) clinical trials comparing extra-amniotic prostaglandin used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This has involved a two-stage method of data extraction. The initial data were extracted centrally, and incorporated into a series of primary reviews arranged by methods of induction of labour, following a standardised methodology. MAIN RESULTS: Oxytocin was used to initiate or augment labour significantly less frequently with extra-amniotic prostaglandins when compared to placebo (relative risk 0.50, 95% confidence interval 0.38-0.66). No other findings were significant in the comparisons that were made for this review including when extra-amniotic prostaglandins were compared with other methods of cervical ripening or induction of labour. Although this could suggest that extra-amniotic prostaglandins are as effective as other agents, the findings are difficult to interpret because they are based on very small numbers and may lack the power to show a real difference. REVIEWER'S CONCLUSIONS: The studies in this review are limited by their small sample sizes which are in many cases further divided into multiple comparison groups. The analyses resulted in most comparisons showing no significant differences, with wide confidence intervals. Although extra-amniotic prostaglandins may be as effective as other modalities in initiating labour, there is little conclusive information from this review to guide clinical practice. An adequately powered randomised controlled trial would be useful to determine if the use of extra-amniotic prostaglandins would lower the rate of caesarean section. However, in the time since these studies were undertaken the use of extra-amniotic prostaglandins has largely been replaced by other modes of prostaglandin administration. PMID- 11406072 TI - Sexual intercourse for cervical ripening and induction of labour. AB - BACKGROUND: The role of prostaglandins for cervical ripening and induction of labour has been examined extensively. Human semen is the biological source that is presumed to contain the highest prostaglandin concentration. The role of sexual intercourse in the initiation of labour is uncertain. The action of sexual intercourse in stimulating labour is unclear, it may in part be due to the physical stimulation of the lower uterine segment, or endogenous release of oxytocin as a result of orgasm or from the direct action of prostaglandins in semen. Furthermore nipple stimulation may be part of the process of initiation. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of sexual intercourse for third trimester cervical ripening or induction of labour in comparison with other methods of induction. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies of relevant papers. Last searched: November 2000. SELECTION CRITERIA: (1) clinical trials comparing sexual intercourse for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusion. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involves a two-stage method of data extraction. MAIN RESULTS: There was one included study of 28 women which reported very limited data, from which no meaningful conclusions can be drawn. REVIEWER'S CONCLUSIONS: The role of sexual intercourse as a method of induction of labour is uncertain. Any future trials investigating sexual intercourse as a method of induction need to be of sufficient power to detect clinically relevant differences in standard outcomes. However, it may prove difficult to standardise sexual intercourse as an intervention to allow meaningful comparisons with other methods of induction of labour. PMID- 11406073 TI - Treatments for iron deficiency anaemia in pregnancy. AB - BACKGROUND: Iron deficiency is the most common cause of anaemia in pregnancy worldwide. Iron treatment can be given by mouth, intramuscular or intravenous injection. Alternatively, blood transfusions and recombinant erythropoietin are also used. OBJECTIVES: To assess the effectiveness of different treatments for iron deficiency anaemia in pregnancy (defined as haemoglobin less than 11 g/dl) on maternal and neonatal morbidity and mortality. SEARCH STRATEGY: Cochrane Pregnancy and Childbirth Group Specialised Register of Trial was searched. Date of last search: December 2000. SELECTION CRITERIA: Randomised controlled trials comparing treatments for iron deficiency anaemia in pregnancy. DATA COLLECTION AND ANALYSIS: The search identified 54 trials. Five trials, involving approximately 1234 women, met the inclusion criteria. Trial quality was assessed. Study authors were contacted for additional information. MAIN RESULTS: Oral iron treatment in pregnancy was assessed in one small trial (n=125), where it was compared with placebo. This showed a reduction in the number of women with haemoglobins under 11g/dl (odds ratio (OR) 0.12, 95% confidence interval (CI) 0.06 to 0.24) and a greater mean haemoglobin level 11.3g/dl compared to 10.5 g/dl (weighted mean difference 0.80, 95% CI 0.62 to 0.98). However, there were no data on clinically relevant outcomes. When comparing different iron treatments, the intravenous (IV) route of administration was associated with an increased risk of venous thrombosis (1 trial, n=74. Iron dextran intramuscularly (IM) versus IV (n=49) OR 0.13, 95% CI 0.02-1.02. IM iron sorbitol-citric acid versus IV iron dextran, OR 0.12, 95% CI 0.02-0.94). Intravenous iron treatments were compared with placebo in one trial (n=54) but only scarce data on adverse outcomes were suitable for inclusion in this review. REVIEWER'S CONCLUSIONS: This review provides inconclusive evidence on the effects of treating iron deficiency anaemia in pregnancy due to the shortage of good quality trials. PMID- 11406074 TI - Hyaluronidase for cervical priming and induction of labour. AB - BACKGROUND: Dilatation and effacement of the cervix are not only a result of uterine contractions, but are also dependent upon ripening processes within the cervix. The cervix is a fibrous organ composed principally of hyaluronic acid, collagen and proteoglycan. Hyaluronic acid increases as pregnancy progresses, increases markedly after the onset of labour and decreases rapidly after birth of the infant. An increase in the level of hyaluronic acid is associated with an increase in tissue water content. Cervical ripening during labour is characterised by changes of the cervix with softening of the tissue and an increased water content. Cervical injection of hyaluronidase was postulated to increase cervical ripening. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of hyaluronidase for third trimester cervical ripening or induction of labour in comparison with other methods of induction of labour SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group Trials Register, the Cochrane Controlled Trials Register and bibliographies of relevant papers. Last searched: November 2000. SELECTION CRITERIA: (1) clinical trials comparing hyaluronidase used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involves a two-stage method of data extraction. MAIN RESULTS: There were no included trials. REVIEWER'S CONCLUSIONS: The role of hyaluronidase as a method of cervical priming or induction of labour is uncertain. PMID- 11406075 TI - Oral prostaglandin E2 for induction of labour. AB - BACKGROUND: This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of oral prostaglandin E2 for third trimester induction of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies of relevant papers. Date of last search: December 2000. SELECTION CRITERIA: The criteria for inclusion included the following: (1) clinical trials comparing oral prostaglandin E2 used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involves a two-stage method of data extraction. The initial data extraction is done centrally, and incorporated into a series of primary reviews arranged by methods of induction of labour, following a standardised methodology. The data will then be extracted from the primary reviews into a series of secondary reviews, arranged by category of woman. To avoid duplication of data in the primary reviews, the labour induction methods have been listed in a specific order, from one to 25. Each primary review includes comparisons between one of the methods (from two to 25) with only those methods above it on the list. MAIN RESULTS: There were 19 studies included in the review. Of these 15 included a comparison using either oral or intravenous oxytocin with or without amniotomy. The quality of studies reviewed was not high. Only seven studies had clearly described allocation concealment. Only two studies stated that providers and/or participants were blinded to treatment group. For the outcome of vaginal delivery not achieved within 24 hours, in the composite comparison of oral PGE2 versus all oxytocin treatments (oral and intravenous, with and without amniotomy), there was a trend favoring oxytocin treatments (relative risk (RR) 1.97, 95% confidence interval (CI) 0.86 to 4.48). For the outcome of cesarean section, in the comparison of PGE2 versus no treatment or placebo, PGE2 was favored (relative risk (RR) 0.54, 95% confidence interval (CI) 0.29,0.98). Otherwise, there were no significant differences between groups for this outcome. Oral prostaglandin was associated with vomiting across all comparison groups. REVIEWER'S CONCLUSIONS: Oral prostaglandin consistently resulted in more frequent gastrointestinal side effects, in particular vomiting, compared with the other treatments included in this review. There were no clear advantages to oral prostaglandin over other methods of induction of labour. PMID- 11406076 TI - Castor oil, bath and/or enema for cervical priming and induction of labour. AB - BACKGROUND: Castor oil, a potent cathartic, is derived from the bean of the castor plant. Anecdotal reports, which date back to ancient Egypt have suggested the use of castor oil to stimulate labour. Castor oil has been widely used as a traditional method of initiating labour in midwifery practice. Its role in the initiation of labour is poorly understood and data examining its efficacy within a clinical trial are limited. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of castor oil or enemas for third trimester cervical ripening or induction of labour in comparison with other methods of cervical ripening or induction of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies of relevant papers. Last searched: November 2000. SELECTION CRITERIA: (1) clinical trials comparing castor oil, bath or enemas used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involves a two-stage method of data extraction. MAIN RESULTS: In the one included study of 100 women, which compared a single dose of castor oil versus no treatment, no difference was found between caesarean section rates (relative risk (RR) 2.31, 95% CI 0.77, 6.87). No data were presented on neonatal or maternal mortality or morbidity. There was no difference between either the rate of meconium stained liquor (RR 0.77, 95% CI 0.25,2.36) or Apgar score < 7 at 5 minutes (RR 0.92, 95% CI 0.02,45.71) between the two groups. The number of participants was small hence only large differences in outcomes could have been detected. All women who ingested castor oil felt nauseous. REVIEWER'S CONCLUSIONS: The only trial included in this review attempts to address the role of castor oil as an induction agent. The trial was small and of poor methodological quality. Further research is needed to attempt to quantify the efficacy of castor oil as an induction agent. PMID- 11406077 TI - Corticosteroids for induction of labour. AB - BACKGROUND: The role of corticosteroids in the process of labour is not well understood. Animal studies have shown the importance of cortisol secretion by the fetal adrenal gland in initiating labour in sheep. Infusion of glucocorticosteroids into the fetus has also shown to induce premature labour in sheep. Some assumptions have been proposed regarding the mode of action of corticosteroids, including both a paracrine and autocrine action, following the identification of glucocorticoid receptors on human amnion. Given these studies it has been postulated that corticosteroids given intraamniotically will promote the induction of labour. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of corticosteroids for third trimester cervical ripening or induction of labour in comparison with other methods of cervical priming or induction of labour. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled Trials Register and bibliographies of relevant papers. Last searched: November 2000. SELECTION CRITERIA: (1) clinical trials comparing corticosteroids used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involves a two-stage method of data extraction. MAIN RESULTS: There are no included trials, hence no results are presented. REVIEWER'S CONCLUSIONS: The effectiveness of corticosteroids for induction of labour is uncertain. Use of this method of induction of labour is not commonly used and further research in this area is probably unwarranted. PMID- 11406078 TI - Vaginal prostaglandin (PGE2 and PGF2a) for induction of labour at term. AB - BACKGROUND: Prostaglandins have been used for induction of labour since the 1960s. Initial work focused on prostaglandin F2a as prostaglandin E2 was considered unsuitable for a number of reasons. With the development of alternative routes of administration, comparisons were made between various formulations of vaginal prostaglandins. This is one of a series of reviews of methods of cervical ripening and labour induction using standardised methodology. OBJECTIVES: To determine the effects of vaginal prostaglandins E2 and F2a for third trimester cervical ripening or induction of labour in comparison with placebo/no treatment or other vaginal prostaglandins (except Misoprostol). SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled trials register and bibliographies of relevant papers. Last searched: November 2000. SELECTION CRITERIA: The criteria for inclusion included the following: (1) clinical trials comparing vaginal prostaglandins used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusions. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involved a two-stage method of data extraction. The initial data extraction was done centrally, and incorporated into a series of primary reviews arranged by methods of induction of labour, following a standardised methodology. The data was then extracted from the primary reviews into a series of secondary reviews, arranged by category of woman. To avoid duplication of data in the primary reviews, the labour induction methods were listed in a specific order, from one to 23. Each primary review included comparisons between one of the methods (from two to 23) with only those methods above it on the list. MAIN RESULTS: In total, 94 studies were considered; 42 have been excluded and 52 included examining a total of 9402 women. Vaginal prostaglandin E2 compared with placebo or no treatment reduced the likelihood of vaginal delivery not being achieved within 24 hours (18% vs. 99%, RR 0.19, 95% CI 0.14,0.25), the caesarean section rates were not different between groups although the risk of uterine hyperstimulation with fetal heart rate changes was increased (4.6% vs. 0.51%, RR 4.14, 95% CI 1.93, 8.90). Comparison of vaginal prostaglandin F2a with placebo showed no increase in caesarean section rates but the cervical score was more likely to be improved (15% vs. 60%, RR 0.25, 95% CI 0.13,0.49), and the risk of oxytocin augmentation reduced (53.9% vs. 89.1%, RR 0.60, 95% CI 0.43,0.84) with the use of vaginal PGF2a. There were insufficient data to make meaningful conclusions for the comparison of vaginal PGE2 and PGF2a. PGE2 tablet, gel and pessary appear to be as efficacious as each other. Lower dose regimes, as defined in the review, appear as efficacious as higher dose regimes. REVIEWER'S CONCLUSIONS: The primary aim of this review was to examine the efficacy of vaginal prostaglandin E2 and F2a. This is reflected by an increase in successful vaginal delivery rates in 24 hours, no increase in operative delivery rates and significant improvements in cervical favourability within 24-48 hours. Further research is needed to quantify the cost-analysis of induction of labour with vaginal prostaglandins, with special attention to different methods of administration. PMID- 11406079 TI - Relaxin for cervical ripening and induction of labour. AB - BACKGROUND: Relaxin is a protein hormone composed of two amino acid chains. The role played by relaxin in human pregnancy and parturition is unclear. Its use and involvement as a cervical ripening agent has been debated since the 1950s. Because the main source of human relaxin is the corpus luteum of pregnancy much of the early work on induction of labour has focused on porcine or bovine preparations. With the advent of DNA recombinant technology human relaxin has become available for evaluation. Relaxin is thought to have a promoting effect on cervical ripening. Due to a possible inhibitory effect on human myometrial activity, relaxin may not be associated with the concomitant increase in the rate of uterine hyperstimulation seen with other induction agents. This is one of a series of reviews of methods of cervical ripening and labour induction using a standardised methodology. OBJECTIVES: To determine the effects of relaxin (both purified porcine and recombinant human) for third trimester cervical ripening or induction of labour in comparison with other methods of induction. SEARCH STRATEGY: The Cochrane Pregnancy and Childbirth Group trials register, the Cochrane Controlled trials register and bibliographies of relevant papers. Last searched: November 2000. SELECTION CRITERIA: (1) clinical trials comparing relaxin used for third trimester cervical ripening or labour induction with placebo/no treatment or other methods listed above it on a predefined list of labour induction methods; (2) random allocation to the treatment or control group; (3) adequate allocation concealment; (4) violations of allocated management not sufficient to materially affect conclusions; (5) clinically meaningful outcome measures reported; (6) data available for analysis according to the random allocation; (7) missing data insufficient to materially affect the conclusion. DATA COLLECTION AND ANALYSIS: A strategy has been developed to deal with the large volume and complexity of trial data relating to labour induction. This involves a two-stage method of data extraction. MAIN RESULTS: In total, nine studies were considered; five have been excluded and four included examining a total of 267 women. There were no reported cases of uterine hyperstimulation with fetal heart rate changes in any of the studies. The rate of caesarean section was not different in those women given relaxin compared with placebo (15.3% versus 14.2%; relative risk (RR) 0.79, 95% confidence interval (CI) 0.42,1.50). There was a reduction in the risk of the cervix remaining unfavourable or unchanged with induction with relaxin (21.9% versus 49.3%; RR 0.45, 95% CI 0.28,0.72). There were no reported cases of uterine hyperstimulation without FHR changes. REVIEWER'S CONCLUSIONS: The place of relaxin, either purified porcine or recombinant human, as an induction or cervical priming agent is unclear. Further trials are needed to estimate the true effect of relaxin within current clinical practice. PMID- 11406080 TI - Advanced trauma life support training for ambulance crews. AB - BACKGROUND: There is an increasing global burden of disease from injuries. Models of trauma care initially developed in high-income countries are also being adopted in low and middle-income countries (LMIC). Amongst these ambulance crews with Advanced Life Support (ALS) training are being promoted in LMIC as a strategy for improving outcomes for victims of trauma. However there is controversy as to the effectiveness of this health service intervention, and the evidence has yet to be rigorously appraised. OBJECTIVES: To quantify the effectiveness of ambulance crews with ALS training versus crews with any other level of training in reducing mortality and morbidity in trauma patients. SEARCH STRATEGY: We searched the Cochrane Controlled Trial Register (CCTR), the specialised register of the Cochrane Injuries Group, MEDLINE, EMBASE, CINAHL, PubMed and the National Research Register. We checked references of background papers and contacted authors to identify additional published or unpublished data. SELECTION CRITERIA: Randomised controlled trials, quasi-randomised controlled trials and controlled before-and-after studies comparing effectiveness of ambulance crews with ALS training versus crews with any other levels of training in reducing mortality and morbidity in trauma patients. Studies which compared crews staffed by physicians versus others were excluded. DATA COLLECTION AND ANALYSIS: Two reviewers independently applied eligibility criteria to trial reports for inclusion and extracted data. MAIN RESULTS: We found one randomised controlled trial, which included 16 trauma cases. However, outcome data were added to the main non-randomised cohort in the analysis, and data on these 16 cases are not yet available. REVIEWER'S CONCLUSIONS: In the absence of evidence of the effectiveness of advanced life support, strong argument could be made that it should not be promoted outside the context of a properly concealed and otherwise rigorously conducted randomised controlled trial. PMID- 11406081 TI - Leadership in academic medicine: a personal perspective. PMID- 11406083 TI - The definitions of acute coronary syndrome, myocardial infarction, and unstable angina. AB - The acute coronary syndrome encompasses a spectrum of conditions that include acute myocardial infarction, unstable angina pectoris, and, to some extent, sudden cardiac death. Recently, the diagnosis of myocardial infarction has been redefined by The Joint European Society of Cardiology/American College of Cardiology Committee. However, the conceptual meaning of the term myocardial infarction has not been changed. Thus, the current diagnoses of myocardial infarction as well as of unstable angina are clinical syndromes based on symptoms, electrocardiogram, and sensitive biochemical markers. PMID- 11406084 TI - Percutaneous coronary intervention in st-elevation myocardial infarction. AB - This review traces the development of coronary angioplasty as a direct, primary means of establishing reperfusion in an acutely occluded infarct-related artery. As a therapeutic strategy for the early management of patients with ST-elevated acute myocardial infarction, this mechanical-invasive procedure arose virtually simultaneously with the demonstration that intravenous administration of fibrinolytic agents to such patients results in a relative reduction in early mortality by as much as 25% to 30%. Over the past 15 years, more than a dozen randomized, controlled trials have investigated the question, "Which is the better means of administrating reperfusion therapy?" The recent demonstration that the combination of coronary stenting plus glycoprotein IIb/IIIa inhibitors used in the early stages of ST-elevation acute myocardial infarction salvages more myocardium than the administration of front-loaded tissue plasminogen activator offers a convincing answer. This is further supported by the evidence that clinical outcomes also improved in these patients, with the greater improvement in ventricular function. It is likely, however, that the development of newer pharmacologic agents plus the combination of both strategies will keep this a hotly debated issue for some time to come. PMID- 11406085 TI - Cardiac markers in the diagnosis of acute coronary syndromes. AB - Evolution of the role of cardiac markers has ranged from the diagnosis of acute myocardial infarction in patients with nondiagnostic electrocardiograms to prognostic risk stratification and to guide therapy. The technology to provide rapid, real time measurements by immunoassay has provided the laboratory and clinician with a range of test options. The principal changes have been the use of rapid serial marker measurements of well-recognized cardiac markers, and the development of immunoassays for the cardiac structural proteins. Measurement of cardiac troponins has generated a new diagnostic paradigm in patients with suspected acute coronary syndromes. There is now a new gold standard biochemical test for myocardial infarction. A range of interventions can be guided by troponin measurement. The use of troponin measurements is central to management of patients with suspected acute coronary syndromes. Future developments in this field will focus on the role of existing and novel markers of inflammation and ischemia. PMID- 11406086 TI - Guidelines for the acute coronary syndromes. AB - Traditional measures employed in the immediate management of patients presenting with an acute coronary syndrome have markedly reduced the risk of early death or myocardial infarction. Further incremental benefit is seen with the substitution of enoxaparin for unfractionated heparin, and in the use of glycoprotein IIb/IIIa inhibitors during percutaneous coronary intervention. However, the evidence for benefit from the glycoprotwin IIb/IIIa inhibitors with medical management alone is unconvincing. Newer data also suggest an aggressive approach to the high-risk patient offers a better ultimate outcome than a conservative one. PMID- 11406088 TI - Should patients with asymptomatic wolff-parkinson-white pattern undergo a catheter ablation? AB - Many individuals with the Wolff-Parkinson-White electrocardiographic pattern are asymptomatic. Optimal management of these individuals is still a matter of debate. On the one hand, sudden cardiac death from ventricular fibrillation is a rare yet possible outcome in these individuals. On the other hand, there is a low risk of serious complication from electrophysiologic study and ablation. Given that the risk of these competing strategies is comparable, the decision needs to be individualized with considerable patient input. PMID- 11406089 TI - Clinical characteristics and catheter ablation of left ventricular outflow tract tachycardia. AB - Left ventricular outflow tract (LVOT) tachycardia is an uncommon form of idiopathic ventricular tachycardia (IVT). The underlying mechanism of this arrhythmia appears to be cyclic AMP-medicated triggered activity. The tachycardia occurs in the absence of structural heart disease and is generally benign, presenting commonly as palpitations and presyncope. It can manifest either a right or left bundle branch block morphology with an inferior axis. Subtle variations in the QRS morphology in leads I, V1, and V2 can help in localizing the anatomic site of origin (SOO). The arrhythmia is typically responsive to a variety of pharmacologic agents (beta-blockers, calcium channel blockers, Class I and II agents). Radiofrequency catheter ablation of LVOT tachycardia SOO as determined by pace mapping is quite efficacious (success rates of 90%). Magnetic electroanatomic mapping augments this by permitting three-dimensional catheter mapping and reproducible localization of the SOO. Catheter ablation should be considered relatively early in patients who experience severe symptoms with their arrhythmia and have failed, or are reluctant to take medications for the disorder. PMID- 11406090 TI - What niche will newer class III antiarrhythmic drugs occupy? AB - The decline in the use of sodium channel blockers has led to an expanding use of b-blockers and complex class III agents such as sotalol and amiodarone for controlling cardiac arrhythmias. Success with these agents in the context of their side effects has spurred the development of compounds with simpler ion channel-blocking properties with less complex adverse reactions. The resulting so called pure class III agents were found to have antifibrillatory effects in atrial fibrillation (AF) and flutter, as well as in ventricular tachyarrhythmias. Pure class III compounds are effective in inducing acute chemical conversion of AF, in preventing paroxysmal AF, and in maintaining sinus rhythm in patients with persistent AF restored to sinus rhythm. Examples of such compounds are dofetilide, which selectively blocks IKr, and ibutilide, available only as an intravenous agent, which blocks the IKr and augments the inactivated Na+ current in atrial myocytes. Dofetilide and ibutilide have been introduced into clinical practice. Azimilide is the first of the class III agents that blocks both components (IKr and IKs) of the delayed rectifier current, which may confer certain electrophysiologic advantages. The potential therapeutic niche of ibutilide, dofetilide, and azimilide in the control of cardiac arrhythmias forms the basis of this review. PMID- 11406091 TI - Advances in device lead extraction. AB - The removal of chronically implanted pacemaker and defibrillator leads has evolved over the past 20 years into an integral part of the medical and surgical practice of electrophysiology. It is the basis for management of device-related complications. Lead removal has progressed from just pulling, to a sophisticated procedure based on telescoping sheaths, counterpressure, and countertraction. Current telescoping sheaths have a powered tip (laser or electrosurgical) for more efficient extirpation of leads from encapsulating fibrous tissue. The management of a device infection exemplifies the spectrum of procedures ranging from debridement of inflammatory tissue to transvenous, transatrial, or epicardial lead reimplantation (care must be given to the consequences of chronic implant complications such as superior vein occlusion). The magnitude of these lead removal procedures ranges from a transvenous procedure to cardiopulmonary bypass removing a pannus of infected material. Current procedure related mortality is less than 0.2% at experienced centers. PMID- 11406092 TI - Evidence for the extracellular reduction of alpha-lipoic acid by Leishmania donovani promastigotes: a transplasma membrane redox system. AB - Leishmania donovani cells, capable of reducing certain electron acceptors with redox potentials at pH 7.0 down to -290 mV, outside the plasma membrane, can reduce the oxidised form of alpha-lipoic acid. alpha-Lipoic acid has been used as natural electron acceptor probe for studying the mechanism of transplasma membrane electron transport. Transmembrane alpha-lipoic acid reduction by Leishmania was not inhibited by mitochondrial inhibitors as azide, cyanide, rotenone or antimycin A, but responded to hemin, modifiers of sulphhydryl groups and inhibitor of glycolysis. The protonophores carbonyl cyanide chlorophenylhydrazone and 2,4-dinitrophenol showed inhibition of alpha-lipoic acid reduction. This transmembrane redox system differs from that of mammalian cells in respect to its sensitivity of UV irradiation and stimulation by diphenylamine. Thus a naphthoquinone coenzyme appears to be involved in alpha lipoic acid reduction by Leishmania cells. PMID- 11406093 TI - Polymorphic phases of galactocerebrosides: spectroscopic evidence of lamellar crystalline structures. AB - Fourier transform infrared spectroscopy was applied to study the structural and thermal properties of bovine brain galactocerebroside (GalCer) containing amide linked non-hydroxylated or alpha-hydroxy fatty acids (NFA- and HFA-GalCer, respectively). Over the temperature range 0-90 degrees C, both GalCer displayed complex thermal transitions, characteristic of polymorphic phase behavior. Upon heating, aqueous dispersions of NFA- and HFA-GalCer exhibited high order-disorder transition temperatures near 80 and 72 degrees C, respectively. En route to the chain melting transition, the patterns of the amide I band of NFA-GalCer were indicative of two different lamellar crystalline phases, whereas those of HFA GalCer were suggestive of lamellar gel and crystalline bilayers. Cooling from the liquid-crystalline phase resulted in the formation of another crystalline phase of NFA-GalCer and a gel phase of HFA-GalCer, with a phase transition near 62 and 66 degrees C, respectively. Prolonged incubation of GalCer bilayers at 38 degrees C revealed conversions among lamellar crystalline phases (NFA-GalCer) or between lamellar gel and crystalline bilayer structures (HFA-GalCer). Spectral changes indicated that the temperature and/or time induced formation of the lamellar crystalline structures of NFA- and HFA-GalCer was accompanied by partial dehydration and by rearrangements of the hydrogen bonding network and bilayer packing mode of GalCer. PMID- 11406094 TI - A functional assay for detection of the mitoxantrone resistance protein, MXR (ABCG2). AB - The fluorescent compounds rhodamine 123, LysoTracker Green DMD-26, mitoxantrone, and BODIPY-prazosin were used with the antagonist fumitremorgin C (FTC) in order to develop functional assays for the half-transporter, MXR/BCRP/ABCP1. A measure of FTC-inhibitable efflux was generated for each compound in a series of MXR overexpressing drug-selected cell lines and in ten unselected cell lines which were used to determine if the four fluorescent compounds were sensitive enough to detect the low MXR levels found in drug-sensitive cell lines. FTC-inhibitable efflux of mitoxantrone and prazosin was found in four of the ten cell lines, SF295, KM12, NCI-H460, and A549, and low but detectable levels of MXR mRNA were also observed by Northern analysis in these cells. FTC-inhibitable mitoxantrone and prazosin efflux in both selected and unselected cell lines was found to correlate well with MXR levels as determined by Northern blotting, r(2)=0.89 and r(2)=0.70 respectively. In contrast, rhodamine and LysoTracker were not able to reliably detect MXR. Cytotoxicity assays performed on two of the four unselected cell lines confirmed increased sensitivity to mitoxantrone in the presence of FTC. FTC was found to be a specific inhibitor of MXR, with half-maximal inhibition of MXR-associated ATPase activity at 1 microM FTC. Short term selections of the SF295, KM12, NCI-H460 and A549 cell lines in mitoxantrone resulted in a small but measurable increase in MXR by both Northern blot and functional assay. These studies show that flow cytometric measurement of FTC inhibitable mitoxantrone or prazosin efflux is a sensitive and specific method for measuring the function of the MXR half-transporter in both selected and unselected cell lines. PMID- 11406095 TI - Differential membrane fluidization by active and inactive cannabinoid analogues. AB - The effects of the two cannabinomimetic drugs (-)-2-(6a,7,10,10a-tetrahydro-6,6,9 trimethyl-1-hydroxy-6H-dibenzo[b,d]pyranyl-2-(hexyl)-1,3-dithiolane (AMG-3) and its pharmacologically less active 1-methoxy analogue (AMG-18) on the thermotropic and structural properties of dipalmitoyl-sn-glycero-3-phosphorylcholine (DPPC) liposomes have been studied by X-ray diffraction and differential scanning calorimetry (DSC). DSC data revealed that the incorporation of the drugs affect differently the thermotropic properties of DPPC. The presence of the more active drug distinctly broadened and attenuated both the pretransition and main phase transition of DPPC bilayers, while the inactive analogue had only minor effects. Small and wide angle X-ray diffraction data showed that the two cannabinoids have different effects on the lipid phase structures and on the hydrocarbon chain packing. The pharmacologically active analogue, AMG-3, was found to efficiently fluidize domains of the lipids in the L(beta)' gel phase, and to perturb the regular multibilayer lattice. In the liquid crystalline L(alpha) phase, AMG-3 was also found to cause irregularities in packing, suggesting that the drug induces local curvature. At the same concentration, the inactive AMG-18 had only minor structural effects on the lipids. At about 10-fold or higher concentrations, AMG 18 was found to produce similar but still less pronounced effects in comparison to those observed by AMG-3. The dose-dependent, different thermotropic and structural effects by the two cannabinoid analogues suggest that these may be related to their biological activity. PMID- 11406096 TI - Protein adsorption to polyethylene glycol modified liposomes from fibrinogen solution and from plasma. AB - Unmodified and polyethylene glycol (PEG) modified neutral and negatively charged liposomes were prepared by freeze-thaw and extrusion followed by chromatographic purification. The effects of PEG molecular weight (PEG 550, 2000, 5000), PEG loading (0-15 mol%), and liposome surface charge on fibrinogen adsorption were quantified using radiolabeling techniques. All adsorption isotherms increased monotonically over the concentration range 0-3 mg/ml and adsorption levels were low. Negatively charged liposomes adsorbed significantly more fibrinogen than neutral liposomes. PEG modification had no effect on fibrinogen adsorption to neutral liposomes. An inverse relationship was found between PEG loading of negatively charged liposomes and fibrinogen adsorption. PEGs of all three molecular weights at a loading of 5 mol% reduced fibrinogen adsorption to negatively charged liposomes. Protein adsorption from diluted plasma (10% normal strength) to four different liposome types (neutral, PEG-neutral, negatively charged, and PEG-negatively charged) was investigated using gel electrophoresis and immunoblotting. The profiles of adsorbed proteins were similar on all four liposome types, but distinctly different from the profile of plasma itself, indicating a partitioning effect of the lipid surfaces. alpha2-macroglobulin and fibronectin were significantly enriched on the liposomes whereas albumin, transferrin, and fibrinogen were depleted compared to plasma. Apolipoprotein AI was a major component of the adsorbed protein layers. The blot of complement protein C3 adsorbed on the liposomes suggested that the complement system was activated. PMID- 11406097 TI - Development of magnetically aligned phospholipid bilayers in mixtures of palmitoylstearoylphosphatidylcholine and dihexanoylphosphatidylcholine by solid state NMR spectroscopy. AB - This study reports the solid-state NMR spectroscopic characterization of a long chain phospholipid bilayer system which spontaneously aligns in a static magnetic field. Magnetically aligned phospholipid bilayers or bicelles are model systems which mimic biological membranes for magnetic resonance studies. The oriented membrane system is composed of a mixture of the bilayer forming phospholipid palmitoylstearoylphosphatidylcholine (PSPC) and the short chain phospholipid dihexanoylphosphatidylcholine (DHPC) that breaks up the extended bilayers into bilayered micelles or bicelles that are highly hydrated (approx. 75% aqueous). Traditionally, the shorter 14 carbon chain phospholipid dimyristoylphosphatidylcholine (DMPC) has been utilized as the bilayer forming phospholipid in bicelle studies. Alignment (perpendicular) was observed with a PSPC/DHPC q ratio between 1.6 and 2.0 slightly above T(m) at 50 degrees C with (2)H and (31)P NMR spectroscopy. Paramagnetic lanthanide ions (Yb(3+)) were added to flip the bilayer discs such that the bilayer normal was parallel with the static magnetic field. The approx. 1.8 (PSPC/DHPC) molar ratio yields a thicker membrane due to the differences in the chain lengths of the DMPC and PSPC phospholipids. The phosphate-to-phosphate thickness of magnetically aligned PSPC/DHPC phospholipid bilayers in the L(alpha) phase may enhance the activity and/or incorporation of different types of integral membrane proteins for solid state NMR spectroscopic studies. PMID- 11406098 TI - Identification and characterization of membrane-associated polypeptides in Torpedo nicotinic acetylcholine receptor-rich membranes by hydrophobic photolabeling. AB - To identify membrane-associated polypeptides present in Torpedo nicotinic acetylcholine receptor (AChR)-rich membranes, we used hydrophobic photolabeling with [(3)H]diazofluorene ([(3)H]DAF) and 1-azidopyrene (1-AP) to tag the membrane proteins which were then identified by amino-terminal sequence analysis of labeled fragments isolated from proteolytic digests by sodium dodecyl sulfate polyacrylamide gel electrophoresis followed by reverse-phase high-performance liquid chromatography. In addition to AChR subunits, identified polypeptides include the 95 kDa alpha-subunit of the (Na(+)+K(+))-ATPase, the 89 kDa voltage gated chloride channel (CLC-0), the 105 kDa SITS-binding protein, and 32 and 34 kDa polypeptides identified as Torpedo homologues of the mitochondrial membrane ATP/ADP carrier protein and the voltage-dependent anion channel (VDAC), respectively. Further, individual amino acids that reacted with [(3)H]DAF and therefore likely to be in contact with lipid were identified in the transmembrane segment M3 of the alpha-subunit of the (Na(+)+K(+))-ATPase and in a putative transmembrane beta-strand in VDAC. Collectively these results demonstrate that [(3)H]DAF/1-AP photolabeling provides an effective method for tagging the membrane-associated segments of polypeptides in a way that makes it easy to isolate the labeled polypeptide or polypeptide fragments by fluorescence and then to identify amino acids at the lipid-protein interface by (3)H release. PMID- 11406099 TI - Na(+)-dependent D-mannose transport at the apical membrane of rat small intestine and kidney cortex. AB - The presence of a Na(+)/D-mannose cotransport activity in brush-border membrane vesicles (BBMV), isolated from either rat small intestine or rat kidney cortex, is examined. In the presence of an electrochemical Na(+) gradient, but not in its absence, D-mannose was transiently accumulated by the BBMV. D-Mannose uptake into the BBMV was energized by both the electrical membrane potential and the Na(+) chemical gradient. D-Mannose transport vs. external D-mannose concentration can be described by an equation that represents a superposition of a saturable component and another component that cannot be saturated up to 50 microM D mannose. D-Mannose uptake was inhibited by D-mannose >> D-glucose>phlorizin, whereas for alpha-methyl glucopyranoside the order was D-glucose=phlorizin >> D mannose. The initial rate of D-mannose uptake increased as the extravesicular Na(+) concentration increased, with a Hill coefficient of 1, suggesting that the Na(+):D-mannose cotransport stoichiometry is 1:1. It is concluded that both rat intestinal and renal apical membrane have a concentrative, saturable, electrogenic and Na(+)-dependent D-mannose transport mechanism, which is different from SGLT1. PMID- 11406100 TI - K(+) transport in red blood cells from human umbilical cord. AB - The current study was designed to characterise K(+) transport in human fetal red blood cells, containing mainly haemoglobin F (HbF, and termed HbF cells), isolated from umbilical cords following normal parturition. Na(+)/K(+) pump activity was comparable to that in normal adult human red cells (which contain HbA, and are termed HbA cells). Passive (ouabain-resistant) K(+) transport was dominated by a bumetanide (10 microM)-resistant component, inhibited by [(dihydroxyindenyl)oxy]alkanoic acid (100 microM), calyculin A (100 nM) and Cl(-) removal, and stimulated by N-ethylmaleimide (1 mM) and staurosporine (2 microM) - all consistent with mediation via the K(+)-Cl(-) cotransporter (KCC). KCC activity in HbF cells was also O(2)-dependent and stimulated by swelling and urea, and showed a biphasic response to changes in external pH. Peak activity of KCC in HbF cells was about 3-fold that in HbA cells. These characteristics are qualitatively similar to those observed in HbA cells, notwithstanding the different conditions experienced by HbF cells in vivo, and the presence of HbF rather than HbA. KCC in HbF cells has a higher total capacity, but when measured at the ambient PO(2) of fetal blood it would be similar in magnitude to that in fully oxygenated HbA cells, and about that required to balance K(+) accumulation via the Na(+)/K(+) pump. These findings are relevant to the mechanism by which O(2) regulates membrane transporters in red blood cells, and to the strategy of promoting HbF synthesis as a therapy for patients with sickle cell disease. PMID- 11406101 TI - Cytoplasmic membrane of a sensitive yeast is a primary target for Cryptococcus humicola mycocidal compound (microcin). AB - A basidiomycetous yeast strain, Cryptococcus humicola 9-6, secretes a mycocidal compound (microcin) which is lethal for many yeasts. In this study a new protocol for microcin purification has been developed, and TLC-purity product was obtained. Using fluorescein as a pH-sensitive probe it was found that microcin treatment of Cryptococcus terreus, a model microcin-sensitive yeast, immediately caused transient alkalization followed by acidification of the cells' cytoplasm. Upon completion of this process, endogenous respiration as well as activity of unspecific esterases were inhibited, and alterations in cell wall and/or capsule started. Microcin was shown to make the cells leaky for intracellular ATP. The mycocidal effect of microcin did not depend on the cell cycle phase of Cr. terreus. Based on these observations and on electrical measurements on planar phospholipid bilayers, which indicated a microcin-induced membrane permeabilization, it is suggested that the cytoplasmic membrane of the sensitive yeast is a primary target of microcin action. The conjectured mode of microcin action involves gradual increase of the cytoplasmic membrane's unspecific permeability. Intracellular ion homeostasis changes induced by microcin are considered to be the main cause of enzyme inhibition, alterations in the outer layers of the cell envelope and, finally, division arrest. PMID- 11406102 TI - Efficient radical trapping at the surface and inside the phospholipid membrane is responsible for highly potent antiperoxidative activity of the carotenoid astaxanthin. AB - The effects of the carotenoids beta-carotene and astaxanthin on the peroxidation of liposomes induced by ADP and Fe(2+) were examined. Both compounds inhibited production of lipid peroxides, astaxanthin being about 2-fold more effective than beta-carotene. The difference in the modes of destruction of the conjugated polyene chain between beta-carotene and astaxanthin suggested that the conjugated polyene moiety and terminal ring moieties of the more potent astaxanthin trapped radicals in the membrane and both at the membrane surface and in the membrane, respectively, whereas only the conjugated polyene chain of beta-carotene was responsible for radical trapping near the membrane surface and in the interior of the membrane. The efficient antioxidant activity of astaxanthin is suggested to be due to the unique structure of the terminal ring moiety. PMID- 11406103 TI - Cationic poly(ethyleneglycol) lipids incorporated into pre-formed vesicles enhance binding and uptake to BHK cells. AB - This paper describes a new method for enhancing the interaction of liposomes with cells. A novel class of cationic poly(ethyleneglycol) (PEG)-lipid (CPL) conjugates have been characterized for their ability to insert into pre-formed vesicles and enhance in vitro cellular binding and uptake of neutral and sterically-stabilized liposomes. The CPLs, which consist of a distearoylphosphatidylethanolamine (DSPE) anchor, a fluorescent dansyl moiety, a heterobifunctional PEG polymer (M(r) 3400), and a cationic headgroup composed of lysine derivatives, have been described previously [Bioconjug. Chem. 11 (2000) 433]. Five separate CPL, possessing 1-4 positive charges in the headgroup (referred to as CPL(1)-CPL(4), respectively), were incubated (as micellar solutions) in the presence of neutral or sterically-stabilized cationic large unilamellar vesicles (LUVs), and were found to insert into the external leaflet of the LUVs in a manner dependent on temperature, time, CPL/lipid ratio, and LUV composition. For CPL/lipid molar ratios < or =0.1, optimal insertion levels of approximately 70% of initial CPL were obtained following 3 h at 60 degrees C. The insertion of CPL resulted in aggregation of the LUVs, as assessed by fluorescence microscopy, which could be prevented by the presence of 40 mM Ca(2+). The effect of CPL-insertion on the binding of LUVs to cells was examined by fluorescence microscopy and quantified by measuring the ratio of rhodamine fluorescence to protein concentration. Neither control LUVs or LUVs containing CPL(2) displayed significant uptake by BHK cells. However, a 3-fold increase in binding was observed for LUVs possessing CPL(3), while for CPL(4)-LUVs values as high as 10 fold were achieved. Interestingly, the increase in lipid uptake did not correlate with total surface charge, but rather with increased positive charge density localized at the CPL distal headgroups. These results suggest that incorporation of CPLs into existing liposomal drug delivery systems may lead to significant improvements in intracellular delivery of therapeutic agents. PMID- 11406104 TI - Na(+)-coupled transport of L-carnitine via high-affinity carnitine transporter OCTN2 and its subcellular localization in kidney. AB - The mechanism of Na(+)-dependent transport of L-carnitine via the carnitine/organic cation transporter OCTN2 and the subcellular localization of OCTN2 in kidney were studied. Using plasma membrane vesicles prepared from HEK293 cells that were stably transfected with human OCTN2, transport of L-carnitine via human OCTN2 was characterized. Uptake of L-[(3)H]carnitine by the OCTN2 expressing membrane vesicles was significantly increased in the presence of an inwardly directed Na(+) gradient, with an overshoot, while such transient uphill transport was not observed in membrane vesicles from cells that were mock transfected with expression vector pcDNA3 alone. The uptake of L-[(3)H]carnitine was specifically dependent on Na(+) and the osmolarity effect showed that Na(+) significantly influenced the transport rather than the binding. Changes of inorganic anions in the extravesicular medium and of membrane potential by valinomycin altered the initial uptake activity of L-carnitine by OCTN2. In addition, the fluxes of L-carnitine and Na(+) were coupled with 1:1 stoichiometry. Accordingly, it was clarified that Na(+) is coupled with flux of L carnitine and the flux is an electrogenic process. Furthermore, OCTN2 was localized on the apical membrane of renal tubular epithelial cells. These results clarified that OCTN2 is important for the concentrative reabsorption of L carnitine after glomerular filtration in the kidney. PMID- 11406105 TI - Regulation of erythrocyte ghost membrane mechanical stability by chlorpromazine. AB - Chlorpromazine (CPZ), a widely used tranquilizer, is known to induce stomatocytic shape changes in human erythrocytes. However, the effect of CPZ on membrane mechanical properties of erythrocyte membranes has not been documented. In the present study we show that CPZ induces a dose-dependent increase in mechanical stability of erythrocyte ghost membrane. Furthermore, we document that spectrin specifically binds to CPZ intercalated into inside-out vesicles depleted of all peripheral proteins. These findings imply that CPZ-induced mechanical stabilization of the erythrocyte ghost membranes may be mediated by direct binding of spectrin to the bilayer. Membrane active drugs that partition into lipid bilayer can thus induce cytoskeletal protein interactions with the membrane and modulate membrane material properties. PMID- 11406106 TI - The structure of the HIV-1 Vpu ion channel: modelling and simulation studies. AB - Vpu is an 81 amino acid auxiliary protein in HIV-1 which exhibits channel activity. We used two homo-pentameric bundles with the helical transmembrane segments derived from FTIR spectroscopy in combination with a global molecular dynamics search protocol: (i) tryptophans (W) pointing into the pore, and (ii) W facing the lipids. Two equivalent bundles have been generated using a simulated annealing via a restrained molecular dynamics simulations (SA/MD) protocol. A fifth model was generated via SA/MD with all serines facing the pore. The latter model adopts a very stable structure during the 2 ns of simulation. The stability of the models with W facing the pore depends on the starting structure. A possible gating mechanism is outlined. PMID- 11406107 TI - Transport of choline and its relationship to the expression of the organic cation transporters in a rat brain microvessel endothelial cell line (RBE4). AB - The present study was undertaken to elucidate the functional characteristics of choline uptake and deduce the relationship between choline uptake and the expression of organic cation transporters in the rat brain microvessel endothelial cell line RBE4. Confluent RBE4 cells were found to express a high affinity choline uptake system. The system is Na(+)-independent and shows a Michaelis-Menten constant of approx. 20 microM for choline. The choline analogue hemicholinium-3 inhibits choline uptake in these cells with an inhibition constant of approx. 50 microM. The uptake system is also susceptible for inhibition by various organic cations, including 1-methyl-4-phenylpyridinium, 1 methyl-4-phenyl-1,2,3,6-tetrahydropyridine, clonidine, procainamide, and tetramethylammonium. The prototypical organic cation tetraethylammonium shows very little affinity for the choline uptake system in these cells. The inhibition of choline uptake by hemicholinium-3 is competitive. Northern analysis and RT-PCR show that these cells do not express the organic cation transporters OCT2 and OCT3. These cells do express, however, low levels of OCT1, but the functional characteristics of choline uptake in these cells are very different from the known properties of choline uptake via OCT1. The Na(+)-coupled high affinity choline transporter CHT1 is not expressed in these cells as evidenced by RT-PCR. This corroborates the Na(+)-independent nature of choline uptake in these cells. It is concluded that RBE4 cells express an organic cation transporter that is responsible for choline uptake in these cells and that this transporter is not identical to any of the organic cation transporters thus far identified at the molecular level in mammalian cells. PMID- 11406108 TI - The interaction of trichosanthin with supported phospholipid membranes studied by surface plasmon resonance. AB - Trichosanthin (TCS) is a toxic protein isolated from a Chinese herbal medicine, the root tuber of Trichosanthes kirilowii Maximowicz of the Curcurbitaceae family. It is now used in China to terminate early and mid-trimester pregnancies. The ribosome inactivating property is thought to be account for its toxicity; it can inactivate the eukaryotic ribosome through its RNA N-glycosidase activity. The interactions of TCS with biological membrane is thought to be essential for its physiological effect, for it must get across the membrane before it can enter the cytoplasm and exert its RIP function. In the present work, the interaction of TCS with supported phospholipid monolayers is studied by surface plasmon resonance. The results show that electrostatic forces dominate the interaction between TCS and negatively charged phospholipid containing membranes under acid condition and that both the pH value and the ionic strength can influence its binding. It is proposed that, besides electrostatic forces, hydrophobic interaction may also be involved in the binding process. PMID- 11406109 TI - Lipid phase separation in phospholipid bilayers and monolayers modeling the plasma membrane. AB - It is postulated that biological membrane lipids are heterogeneously distributed into lipid microdomains. Recent evidence indicates that docosahexaenoic acid containing phospholipids may be involved in biologically important lipid phase separations. Here we investigate the elastic and thermal properties of a model plasma membrane composed of egg sphingomyelin (SM), cholesterol and 1-stearoyl-2 docosahexaenoyl-sn-glycerophosphoethanolamine (SDPE). Two techniques are employed, pressure-area isotherms on monolayers to examine condensation and interfacial elasticity behavior, and differential scanning calorimetry (DSC) on bilayers to evaluate phase separations. Significant levels of condensation are observed for mixtures of SM and cholesterol. Surface elasticity measurements indicate that cholesterol decreases and SDPE increases the in-plane elasticity of SM monolayers. At X(SDPE)> or =0.15 in SM, a more horizontal region emerges in the pressure-area isotherms indicating 'squeeze out' of SDPE from the monolayers. Addition of cholesterol to equimolar amounts of SM and SDPE further increases the amount of 'squeeze out', supporting the concept of phase separation into a cholesterol- and SM-rich liquid ordered phase and a SDPE-rich liquid disordered phase. This conclusion is corroborated by DSC studies where as little as X(Chol)=0.0025 induces a phase separation between the two lipids. PMID- 11406110 TI - The influence exerted by the beta(3) subunit on MVIIA omega-conotoxin binding to neuronal N-type calcium channels. AB - In the present study, two-electrode voltage-clamp techniques have been used to assess the interaction between the MVIIA omega-conotoxin and an isoform of the N type Ca(2+) channel alpha subunit (alpha(1B-d)). Cloned alpha(1B-d) Ca(2+) channels were expressed in Xenopus laevis oocytes in the presence and absence of the beta(3) subunit. Coexpression of the beta(3) subunit significantly shifted the IC(50) value for MVIIA inhibition of central N-type Ca(2+) channel current. Analysis of the peak conductance vs. depolarising voltage dependence suggested that the beta(3) subunit has no apparent effect on the gating charge which accompanies the closed-open transition of the channels. Instead, coexpression of the beta(3) subunit led to an approx. 10 mV shift to more hyperpolarised potentials in the voltage-dependent activation of N-type Ca(2+) channels. We conclude that MVIIA alters the surface charge on the N-type Ca(2+) channels and might induce allosteric changes on the structure of the channel, leading to an increase in the dissociation constant of MVIIA binding. PMID- 11406111 TI - Human cystine/glutamate transporter: cDNA cloning and upregulation by oxidative stress in glioma cells. AB - A human cDNA for amino acid transport system x(C)(-) was isolated from diethyl maleate-treated human glioma U87 cells. U87 cells expressed two variants of system x(C)(-) transporters hxCTa and hxCTb with altered C-terminus regions probably generated by the alternative splicing at 3'-ends. Both hxCTa and hxCTb messages were also detected in spinal cord, brain and pancreas, although the level of hxCTb expression appears to be lower than that of hxCTa in these tissues. When expressed in Xenopus oocytes, hxCTb required the heavy chain of 4F2 cell surface antigen (4F2hc) and exhibited the Na(+)-independent transport of L cystine and L-glutamate, consistent with the properties of system x(C)(-). In agreement with this, 137 kDa band was detected by either anti-xCT or anti-4F2hc antibodies in the non-reducing condition in western blots, whereas it shifted to 50 kDa or 90 kDa bands in the reducing condition, indicating the association of two proteins via disulfide bands. We found that the expression of xCT was rapidly induced in U87 cells upon oxidative stress by diethyl maleate treatment, which was accompanied by the increase in the L-cystine uptake by U87 cells. Because of this highly regulated nature, xCT in glial cells would fulfill the task to protect neurons against oxidative stress by providing suitable amount of cystine to produce glutathione. PMID- 11406112 TI - Effect of cholesterol on miscibility and phase behavior in binary mixtures with synthetic ceramide 2 and octadecanoic acid. Infrared studies. AB - The three main lipid components of the stratum corneum, namely ceramides, free fatty acids and cholesterol, play a fundamental role in the maintenance of the skin barrier. The current investigation is aimed toward understanding the miscibility and intermolecular interactions of these lipids. Toward this end, Fourier transform infrared spectroscopic studies of the three possible equimolar binary mixtures of cholesterol, a synthetic non-hydroxylated fatty acid N-acyl sphingosine with a C18 chain length (N-stearoylsphingosine, approximating human ceramide 2), and stearic acid were undertaken. The thermotropic responses of the methylene stretching and scissoring vibrations were used to evaluate chain conformation and packing respectively. Selective perdeuteration, of either the stearic acid or the ceramide acid chains, permitted separate and simultaneous evaluation of the conformational order and packing properties of the sphingosine chain, the amide linked fatty acid chains and/or the stearic acid chain. Whereas cholesterol mixed well with ceramide at physiological temperatures, the stearic acid was miscible with the cholesterol only at relatively high temperatures where the fatty acid is disordered. A complex interaction between stearic acid and ceramide was detected. A separate fatty acid-rich phase persisted until at least 50 degrees C, whereas at higher temperatures the components appear to be quite miscible. However, a preferential association of the fatty acid with the ceramide base chain is indicated. None of the binary systems studied exhibit miscibility and interactions resembling those in the ternary mixtures of these substances, which is widely used to model stratum corneum. The role of cholesterol in controlling the miscibility characteristics in the ternary system is evident. PMID- 11406113 TI - Iron-induced oxidative damage of corn root plasma membrane H(+)-ATPase. AB - The effect of iron on the activity of the plasma membrane H(+)-ATPase (PMA) from corn root microsomal fraction (CRMF) was investigated. In the presence of either Fe(2+) or Fe(3+) (100-200 microM of FeSO(4) or FeCl(3), respectively), 80-90% inhibition of ATP hydrolysis by PMA was observed. Half-maximal inhibition was attained at 25 microM and 50 microM for Fe(2+) and Fe(3+), respectively. Inhibition of the ATPase activity was prevented in the presence of metal ion chelators such as EDTA, deferoxamine or o-phenanthroline in the incubation medium. However, preincubation of CRMF in the presence of 100 microM Fe(2+), but not with 100 microM Fe(3+), rendered the ATPase activity (measured in the presence of excess EDTA) irreversibly inhibited. Inhibition was also observed using a preparation further enriched in plasma membranes by gradient centrifugation. Addition of 0.5 mM ATP to the preincubation medium, either in the presence or in the absence of 5 mM MgCl(2), reduced the extent of irreversible inhibition of the H(+)-ATPase. Addition of 40 microM butylated hydroxytoluene and/or 5 mM dithiothreitol, or deoxygenation of the incubation medium by bubbling a stream of argon in the solution, also caused significant protection of the ATPase activity against irreversible inhibition by iron. Western blots of CRMF probed with a polyclonal antiserum against the yeast plasma membrane H(+)-ATPase showed a 100 kDa cross-reactive band, which disappeared in samples previously exposed to 500 microM Fe(2+). Interestingly, preservation of the 100 kDa band was observed when CRMF were exposed to Fe(2+) in the presence of either 5 mM dithiothreitol or 40 microM butylated hydroxytoluene. These results indicate that iron causes irreversible inhibition of the corn root plasma membrane H(+)-ATPase by oxidation of sulfhydryl groups of the enzyme following lipid peroxidation. PMID- 11406114 TI - Cloning, cellular distribution and functional expression of small intestinal epithelium guinea pig ClC-5 chloride channel. AB - We report the cloning of a guinea pig ClC-5 chloride channel (gpClC-5) from distal small intestinal epithelial cells by RT-PCR and RACE. The transcript is shown to be present in duodenum, jejunum and ileum epithelium by RT-PCR and Northern analysis. This is confirmed by in situ hybridisation which also shows the transcript to be homogeneously distributed in the crypt and villus regions. Expression of gpClC-5 in HEK-293 cells generated markedly outwardly rectified chloride currents with a perm-selectivity sequence of NO(3)(-)>Cl(-)>Br(-)>I( )>F(-)>gluconate(-). The possible role of gpClC-5 in this epithelial location is discussed. PMID- 11406116 TI - Presenilin 1 mutations increase amyloid precursor protein production and proteolysis in Xenopus laevis oocytes. AB - Recent findings suggest that Presenilin 1 (PS1) mutations play a major role in the development of Alzheimer's disease (AD) by increasing the production of the beta amyloid peptide (A beta). The exact mechanism whereby mutations in PS1 lead to this effect is not clear. To examine the question of how PS1 might be involved in amyloid precursor protein (APP) processing, we constructed a chimera of human APP695 fused at the C-terminal to enhanced green fluorescent protein (EGFP). This construct was injected into Xenopus laevis oocytes in the presence of wild type PS1 or one of three PS1 mutations associated with AD. The cellular location of the APP695-EGFP construct was examined by fluorescent confocal microscopy. In addition, membrane fractions of oocytes expressing APP695-EGFP in the presence or absence of wild type or mutant forms of PS1 were evaluated by Western blot analysis. The results show that APP695-EGFP is primarily expressed on the cell surface and undergoes limited cleavage. Specifically, APP695 was cleaved in the A beta domain to generate three distinct C-terminal fragments that correspond in length to stubs expected after cleavage with alpha-, beta- and gamma-secretase, respectively. The presence of wild type PS1 not only increased the production of proteolytic C-terminal fragments of APP, but the production of APP itself. These findings were even more pronounced in the presence of all three PS1 mutations, suggesting that PS1 mutations may lead to over-expression of APP not just increased gamma-secretase activity. PMID- 11406117 TI - Fractionated radiation facilitates repair and functional motor recovery after spinal cord transection in rat. AB - Previous studies suggest that motor recovery does not occur after spinal cord injury because reactive glia abort the natural repair processes. A permanent wound gap is left in the cord and the brain-cord circuitry consequently remains broken. Single-dose x-irradiation destroys reactive glia at the damage site in transected adult rat spinal cord. The wound then heals naturally, and a partially functional brain-cord circuitry is reconstructed. Timing is crucial; cell ablation is beneficial only within the third week after injury. Data presented here point to the possibility of translating these observations into a clinical therapy for preventing the paralysis following spinal cord injury in the human. The lesion site (at low thoracic level) in severed adult rat spinal cord was treated daily, over the third week postinjury, with protocols of fractionated radiation similar to those for treating human spinal cord tumors. This resulted, as with the single-dose protocol, in wound healing and restoration of some hindquarter motor function; in addition, the beneficial outcome was augmented. Of the restored hindlimb motor functions, weight-support and posture in stance was the only obvious one. Recovery of this motor function was partial to substantial and its incidence was 100% instead of about 50% obtained with the single-dose treatment. None of the hindlimbs, however, regained frequent stepping or any weight-bearing locomotion. These data indicate that the therapeutic outcome may be further augmented by tuning the radiation parameters within the critical time window after injury. These data also indicate that dose-fractionation is an effective strategy and better than the single-dose treatment for targeting of reactive cells that abort the natural repair, suggesting that radiation therapy could be developed into a therapeutic procedure for repairing injured spinal cord. PMID- 11406118 TI - Reactive nitrogen species contribute to blood-labyrinth barrier disruption in suppurative labyrinthitis complicating experimental pneumococcal meningitis in the rat. AB - Sensorineural hearing damage is a frequent complication of bacterial meningitis, affecting as many as 30% of survivors of pneumococcal meningitis. There is a substantial body of evidence that oxidants, such as reactive nitrogen species (RNS), are central mediators of brain damage in experimental bacterial meningitis. In the present study, we investigated whether RNS also contribute to the pathophysiology of suppurative labyrinthitis in our well-established rat model of pneumococcal meningitis. In all infected rats, but not in uninfected controls, we observed suppurative labyrinthitis. Cochlear inflammation was accompanied by severe blood-labyrinth barrier (BLB) disruption as evidenced by increased Evans Blue extravasation. Furthermore, increased cochlear expression of endothelial nitric oxide synthase (eNOS) and inducible nitric oxide synthase (iNOS) was detected by immunohistochemistry. Colocalization of iNOS and tyrosine nitration (a marker of RNS attack) indicated that nitric oxide (NO) produced by iNOS contributes to oxidative cochlear damage through the action of RNS. To determine the pathophysiological role of RNS in BLB disruption, rats were treated with peroxynitrite scavengers (MnTBAP and uric acid, UA). Six h after adjunctive treatment with 300 mg/kg i.p. UA or 15 mg/kg i.p. MnTBAP+100 mg/kg i.p. ceftriaxone, BLB disruption was significantly reduced compared with that in infected animals treated only with ceftriaxone. Therefore, we conclude that RNS are involved in the breaching of the BLB during meningogenic pneumococcal labyrinthitis. PMID- 11406119 TI - The effect of feeding on cerebrospinal fluid corticotropin-releasing hormone levels in humans. AB - Corticotropin-releasing hormone (CRH) is a neuropeptide thought to play a role in appetite regulation. In this report, we used a serial cerebrospinal fluid (CSF) sampling technique to examine the relationship between CSF CRH, plasma ACTH and cortisol and perceptions of hunger and satiety in fasting and sated volunteers. CSF was withdrawn continuously from 11:00 AM to 5:00 PM via an indwelling subarachnoid catheter. Blood was withdrawn every 10 min via an antecubital vein catheter. Fed subjects received a meal at 1:00 PM. Subjects who were fed had lower post-prandial ratings on hunger scales and higher ratings on satiety scales. Fed subjects also had slightly lower levels of CSF CRH after feeding. Furthermore, fed subjects had higher ACTH and cortisol concentrations in the first 3 h; by the fourth h the opposite was true. Our findings do not support the hypothesis that CNS CRH is a central satiety factor in the human. Instead our findings of slightly diminished CSF CRH levels after feeding may be accounted for by the rises in glucocorticoids and their associated negative feedback effects on CNS CRH. Alternatively, our findings could also reflect changes in CRH levels associated with feeding in multiple brain areas and in the spinal cord with the net effect being in the negative direction. PMID- 11406120 TI - Long-lasting handling affects behavioural reactivity in adult rats of both sexes prenatally exposed to diazepam. AB - Environmental stressors can substantially affect the adaptive response of rats to novelty in a sexually dimorphic manner. Gender-related differences are also observed in neurochemical and behavioural patterns of adult rats following prenatal exposure to diazepam (DZ). In the present study the behavioural reactivity to novelty is investigated in open field (OF) and in acoustic startle reflex (ASR) tests, in non handled (NH), short-lasting handled (SLH) and long lasting handled (LLH) adult male and female rats prenatally exposed to DZ. A single daily s.c. injection of DZ (1.5 mg/kg) over gestation days 14-20 decreases GABA/BDZ receptor function in both sexes, as shown by the decreased electrographic hippocampal response to DZ and the increased response to picrotoxin, after intra-locus coeruleus injection of the two compounds. In OF NH DZ-exposed males display a lower total distance travelled (TDT), a higher rearing frequency (RF) and a greater number of transitions in the centre of the arena (CNT) compared to NH rats prenatally exposed to vehicle. Conversely, NH DZ exposed females show slight changes in TDT and RF and a greater reduction in CNT and in the amount of time spent in the centre of the arena (CAT). These effects are associated with an increase in the peak amplitude of the ASR in both sexes. Short-lasting handling slightly influences DZ-evoked effects in animals of both sexes. In DZ-exposed males long-lasting handling attenuates the reduction in TDT and the enhancement in RF, prevents the increase in CNT and reduces the peak amplitude of ASR. In DZ-exposed females, long-lasting handling increases TDT and RF, induces a lower avoidance of the centre of the arena, and does not modify the peak amplitude of ASR, when compared to controls. These findings indicate that prenatal exposure to DZ differently affects behavioural reactivity in adult male and female rats, and suggest that a long-lasting handling is able to attenuate some behavioural deficits induced by prenatal DZ exposure. PMID- 11406121 TI - Orexin A-like immunoreactivity in the hypothalamus and thalamus of the Syrian hamster (Mesocricetus auratus) and Siberian hamster (Phodopus sungorus), with special reference to circadian structures. AB - The orexins are recently discovered neuropeptides that reportedly play a role in energy homeostasis, in addition to various other physiological processes. The synthesis of orexin A undergoes diurnal variation in certain areas of the brain, while the mutation of the orexin receptor 2 gene has been implicated in canine narcolepsy. Since the circadian pacemaker in the suprachiasmatic nucleus modulates the sleep/wake cycle, there is a putative role for orexins in the mammalian circadian system. In this study, immunohistochemical techniques were used to determine the distribution of orexin A in the structures of the hypothalamus and thalamus of Syrian and Siberian hamsters. In both species, the pattern of immunoreactivity was similar. Cells immunoreactive for orexin A were noted in the lateral hypothalamic area. Immunoreactive varicose orexin A fibres were found throughout the hypothalamus. The suprachiasmatic nucleus possessed little or no immunoreactive orexin A fibres in its core, but had fibres at its periphery. The thalamus of both species contained comparatively few immunoreactive fibres, which were mainly localised around the midline. The thalamic intergeniculate leaflet contained a plexus of immunoreactive orexin A fibres throughout its rostro-caudal extent. Three areas of the brainstem, the dorsal and median raphe nuclei and the locus coeruleus, were also investigated owing to their relevance to the circadian system and all were found to contain immunoreactive orexin A fibres. The presence of orexin A-immunoreactive fibres in the neural architecture of the mammalian circadian system suggests an important role for orexin A in circadian timekeeping processes. PMID- 11406122 TI - Neuroprotection of S(+) ketamine isomer in global forebrain ischemia. AB - The non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist ketamine can block the action of excitotoxic amino acids in the central nervous system. S(+) ketamine has a 2-3 times higher anesthetic potency compared with the ketamine racemate and also shows a higher neuroprotective efficacy in vitro. To determine the neuroprotective activity of S(+) ketamine compared with its R(-) stereoisomer in vivo, we examined the functional and neurohistological outcome in rats treated 15 min after global forebrain ischemia with S(+) ketamine in different dosages compared with R(-) ketamine. Influence of the treatment on regional cerebral blood flow (rCBF) and cortical oxygen saturation (HbO2) was monitored over 1 h after the ischemia using laser doppler flowmetry and microphotospectrometry respectively. Sixty and ninety mg/kg of S(+) ketamine but not R(-) ketamine significantly reduced neuronal cell loss in the cortex compared with the saline treated group. No significant neuroprotection was observed in the hippocampus. Although no significant change in rCBF was found, S(+) ketamine restored the cortical HbO2 to preischemic values. These results indicate that S(+) ketamine in higher dosages can reduce neuronal damage in the cortex after cerebral ischemia, possibly by improving the ratio of oxygen supply to consumption in the postischemic tissue. PMID- 11406123 TI - Possible mechanism of hypothermia induced by intracerebroventricular injection of orphanin FQ/nociceptin. AB - Orphanin/nociceptin (OFQ/N), a 17-amino-acid peptide, is an endogenous peptide, the receptor for which is similar to mu-, delta- and kappa-opioid receptors ( approximately 65% homology). Reports indicate that OFQ/N can block the antinociception induced by mu-, delta- and kappa-opioid agonists in the rat and in the mouse, indicating that there is a functional interaction between opioid receptors and OFQ/N receptors in the nervous system. It is well known that activation of the mu- and kappa-opioid receptors results in hyperthermia and hypothermia, respectively, in Sprague-Dawley rats. The present studies were designed to examine effects of OFQ/N on body temperature (Tb) and explore whether the mechanism of T(b) change induced by OFQ/N involved the opioid system. The results show that (1) i.c.v. injection of a high dose of OFQ/N (9-18 micro g) produces hypothermia in adult rats; (2) OFQ/N (1.8 micro g, i.c.v., t=+30 s after morphine) can decrease morphine-induced hyperthermia; (3) neither the opioid receptor antagonist, naloxone (10 mg/kg, s.c., t=-15 s before OFQ/N) nor the kappa-opioid receptor antagonist nor-BNI (1 micro g/5 microl, i.c.v., t=-30 s before OFQ/N) reduces the hypothermia induced by i.c.v. injection of OFQ/N at dose of 18 micro g (P>0.05); (4) 60 micro g/5 microl AS oligo (i.c.v. treatment on days 1, 3 and 5) against OFQ/N receptors significantly reduces the hypothermia induced by i.c.v. injection of 9 micro g OFQ/N (P<0.01). These results suggest that the hypothermia induced by i.c.v. injection of a high dose of OFQ/N (9 or 18 micro g) is mediated, at least partially, by its own receptor, independent or downstream of opioid receptors in the rat brain and that OFQ/N probably acts as a physiological antagonist to reduce morphine-induced hyperthermia. PMID- 11406124 TI - Nuclear translocation of STAT5 and increased expression of Fos related antigens (FRAs) in hypothalamic dopaminergic neurons after prolactin administration. AB - Ample evidence indicates feedback relationships between pituitary prolactin and hypothalamic dopaminergic neurons. Since the presence of prolactin receptors was earlier demonstrated in hypothalamic dopaminergic neurons, our working hypothesis was that prolactin induced activation of prolactin receptor coupled signaling leads to increased neuronal activity in these neurons. The aim of this study was to correlate prolactin receptor mediated signaling and prolactin induced activation in hypothalamic dopaminergic neurons. We used nuclear translocation of STAT5 as a marker of prolactin receptor induced signaling and expression of Fos related antigens (FRAs) as an indicator of neuronal activation. We performed double label immunocytochemical studies to determine the time course of the presence of FRAs and STAT5 in the nuclei of hypothalamic dopaminergic neurons after ovine prolactin treatment. Exogenous ovine prolactin treatment of ovariectomized rats resulted in an increase in serum ovine prolactin levels and a decrease in endogenous serum prolactin levels, indicating that ovine prolactin activated mechanisms inhibited pituitary prolactin secretion. Indeed, ovine prolactin activated the prolactin receptors in most subpopulations of hypothalamic dopaminergic neurons, resulting in nuclear translocation of STAT5. Also, increased neuronal activity, indicated by expression of FRAs, was observed in the same neuron populations after ovine prolactin treatment. These results suggest that signal transduction mechanisms coupled to prolactin receptors in hypothalamic dopaminergic neurons resemble those observed in other tissues; and nuclear translocation of STAT5 can be used as a marker of prolactin receptor activation in hypothalamic dopaminergic neurons. PMID- 11406125 TI - Involvement of c-Jun N-terminal kinase and caspase 3-like protease in DNA damage induced, p53-mediated apoptosis of cultured mouse cerebellar granule neurons. AB - In the previous studies, we have demonstrated that the tumor suppressor gene p53 is required for DNA strand break-induced neuronal apoptosis in organotypic slice cultures of cerebellum as well as in dissociated cerebellar neuron cultures. In this study, we further investigated the role of p53 in neuronal apoptosis, by examining whether caspases and c-Jun N-terminal kinase (JNK) are involved in the DNA strand break-induced apoptosis. The protein level of phospho-JNK increased in p53 wild-type mouse cerebellar granule neurons after exposure to bleomycin. On the other hand, the response was not observed in cerebellar granule neurons of p53-deficient mice. Caspase-3-like protease was activated and poly(ADP-ribose) polymerase (PARP) was cleaved in the bleomycin-induced apoptosis. Caspase-3-like protease inhibitor decreased the number of TUNEL-positive but not p53- or c-Jun positive neurons in bleomycin-induced death. These results suggest that JNK and caspase-3-like protease are involved in the signaling cascade of DNA strand break induced, p53-dependent apoptosis. PMID- 11406126 TI - Chronic restraint stress enhances radial arm maze performance in female rats. AB - Effects of chronic restraint stress (21 and 28 days) on physiological and behavioral parameters in female rats were examined. Total (bound and free) and free corticosterone (CORT) levels were measured at different time points during the stress period. Higher total CORT levels were observed in stressed rats during the stress period but returned to baseline at 15 days post-stress. Additionally, free CORT levels decreased across the stress period. Estrous cyclicity was monitored daily in all animals. Stress had no apparent effects on estrous cyclicity, in rats with either normal length or elongated estrous cycles, but stressed females gained less weight than controls. Following the stress period, subjects were tested for open field activity and radial arm maze (RAM) performance. Females stressed for 21 days showed enhanced spatial memory performance on the RAM. A longer period of restraint, 28 days, also led to less weight gain by stressed subjects and unaltered estrous cycle lengths, but was not associated with enhanced RAM performance. Further analysis indicated that RAM performance was influenced by specific estrous cycle day, particularly during proestrus. Following 21 days of restraint stress all animals in proestrus, regardless of treatment, showed impaired acquisition. After 28 days, stressed females in proestrus performed better than proestrus controls. These results are discussed in relation to previously reported effects of stress in male rats. PMID- 11406127 TI - Effect of embryonic knock-down of GABAA receptors on the levels of monoamines and their metabolites in the CNS of the mouse. AB - In vitro evidence indicates that gamma-aminobutyric acid (GABA), acting at GABA(A) receptors, exerts a positive trophic effect on monoaminergic neurons during embryogenesis. To determine whether in vivo antagonism of GABA(A) receptors during embryogenesis interferes with the development of monoaminergic neurons, we used mice in which the number of GABA(A) receptors was decreased by 50% by targeted deletion of the beta(3) subunit gene of the GABA(A) receptor. Levels of serotonin, dopamine, norepinephrine, and the metabolites 3,4 deoxyphenylacetic acid, homovanillic acid, and 5-hydroxyindoleacetic acid were measured in the brainstem, cortex, striatum and spinal cord of female adult homozygous null (beta3-/-) and wild-type (beta3+/+) mice, as well as progenitor C57BL/6J and Strain 129/SvJ mice. The level of norepinephrine in the spinal cord of beta3-/- mice was 44% less than that of beta3+/+ mice, and did not differ in the brainstem, cortex or striatum. This finding suggests that beta3 subunit containing GABA(A) receptors mediate the trophic effects of GABA on a subpopulation of spinally-projecting noradrenergic neurons. In contrast, the levels of serotonin, dopamine or their metabolites were unaffected, suggesting that the development of serotonergic and dopaminergic neurons may require activation of only a small fraction of GABA(A) receptors or may not be dependent on beta3 subunit-containing GABA(A) receptors. Finally, Strain 129/SvJ and C57BL/6J mice differed with respect to the levels of dopamine and its metabolites in the brainstem, spinal cord and cortex. These differences may need to be considered when assessing the phenotype of gene-targeted mice for which these mice serve as progenitor strains. PMID- 11406128 TI - Behavioural disturbances and sensory pathology following allylnitrile exposure in rats. AB - Animals exposed to allylnitrile develop permanent abnormalities in motor behaviour, similar to those caused by 3,3'-iminodipropionitrile (IDPN) and crotononitrile. IDPN and crotononitrile effects have been attributed to vestibular hair cell degeneration, but allylnitrile has been suggested to modify behaviour through neuronal degeneration in the CNS. Adult male Long-Evans rats were exposed to allylnitrile (0, 20, 40, 60 mg/kg per day, for 3 days) and the changes in rearing activity and rating scores in tests of vestibular function were assessed. Surface preparations of the vestibular sensory epithelia and the organ of Corti were observed for hair cell loss by scanning electron microscopy. Corneal transparency and concentrations in retina and olfactory bulbs of glial fibrillary acidic protein (GFAP), a marker for reactive gliosis, were also determined, as they are known targets of IDPN toxicity. In a dose-dependent manner, allylnitrile caused corneal opacity and gliosis in the retina and olfactory bulbs, decreased rearing activity and increased the rating scores in tests of vestibular dysfunction, and induced hair cell loss in both the vestibular sensory epithelia and the organ of Corti. The behavioural deficits correlated well with the loss of vestibular hair cells. We conclude that allylnitrile causes permanent modifications in behaviour by loss of vestibular function as IDPN and crotononitrile do and that all these chemicals share other toxic targets, such as the cornea, the retina, and the olfactory system. Data reported here and elsewhere indicate that a number of nitriles show similar neurotoxic properties. PMID- 11406129 TI - ATP modulates Na+ channel gating and induces a non-selective cation current in a neuronal hippocampal cell line. AB - Extracellular ATP evoked two excitatory responses in hippocampal neuroblastoma cells (HN2). The first, an opening of a receptor-operated non-selective cation channel and the second was a leftward shift in Na+ channel activation. Both ATP (5-1000 microM) and 2',3'-(4-benzoyl)-benzoyl-ATP (Bb-ATP, 50 microM) activated a non-selective cation current reversing near 0 mV and shifted the Na+ activation and inactivation curves to the left. Based on a comparison of a series of agonists and antagonists, the inward current appeared to be partially mediated by activation of a P2X7 receptor, although hybrid channels cannot be ruled out. The shift in Na+ channel gating could be separated from the opening of the cation channel, as application of the P2Y antagonist Reactive Blue-2 and GTP shifted the Na+ current activation to the left but failed to elicit the inward cation current. Both responses to ATP and Bb-ATP were insensitive to block by the P2X antagonist suramin (300 microM) but were prevented by incubation in oxidized ATP (200 microM); a putative P2X7 receptor antagonist. Prior screening of the surface negative charge of the membrane with a high concentration of divalent cations prevented both responses. We suggest that ATP4- activates a P2X receptor and becomes trapped on a site, on or near the Na+ channel. Activation of the P2X receptor leads to the opening of a non-specific cation channel, while the binding of ATP4- leads to a modified charge sensed by the Na+ channel, similar to what occurs in the presence of charged amphiphiles as well as a number of beta scorpion toxins. PMID- 11406130 TI - Anatomical and ontogenetic factors producing variation in HVc neuron number in zebra finches. AB - The volume of nucleus HVc of the avian song system varies greatly both among and within songbird species, and is positively correlated to song complexity in many species. Moreover, the number of neurons in HVc predicts the ability of individual zebra finches to imitate song accurately. To better understand this brain/behavior relationship, we used the retrograde tracer Fast Blue to assess how specific HVc neuronal subpopulations contribute to variation in overall HVc neuron number in adult male zebra finches. We also investigated whether sibling order predicts the number of HVc neurons and/or yolk levels of testosterone, a hormone that might regulate the development of HVc. We report that total HVc neuron number is consistently and independently predicted by the size of each of its two projection populations, suggesting that the proportional makeup of HVc is tightly regulated at least in male zebra finches. Also, while we failed to detect a significant effect of sibling order on either the number of neurons in HVc or yolk testosterone concentration, we found that clutch of origin made a large contribution to variation in both early hormone levels and HVc neuron number. PMID- 11406131 TI - The effects of lead on transient outward currents of acutely dissociated rat dorsal root ganglia. AB - The effects of Pb2+ on transient outward currents (TOCs) were investigated on rat dorsal root ganglia (DRG) neurons at postnatal days of 15 approximately 21, using the conventional whole-cell patch-clamp technique. In media-sized (35 approximately 40 microm) neurons and in the presence of 50 mM TEA, TOCs that preliminarly included an A-current (IA) and a D-current (ID), were clearly present and dominant. Application of Pb2+ lengthened the initial delay of TOCs and increased the onset-peak time in a concentration-dependent manner. The amplitudes of initial outward current peak were reduced with increasing Pb2+ concentrations. The inhibitory effects of Pb2+ on TOCs were reversible with 80 approximately 90% of current reversed in 2 approximately 10 min at 1 approximately 400 microM Pb2+. For the normalized activation curves fitted by a single Boltzmann equation under each condition, there was a shift to more depolarized voltages with increasing concentrations of Pb2+. The V1/2 and the slope factor (k) increased from 12.76+/-1.49 mV and 15.31+/-1.66 mV (n=10) under control condition to 39.91+/-5.44 mV (n=10, P<0.01) and 21.39+/-3.13 mV (n=10, P<0.05) at 400 microM Pb2+, respectively, indicating that Pb2+ decreased the activation of TOCs. For the normalized steady-state inactivation curves, the V1/2 and the k increased from -92.31+/-2.72 and 8.59+/-1.36 mV (n=10) to -55.65+/-3.67 (n=10, P<0.01) and 23.02+/-2.98 mV (n=10, P<0.01) at 400 microM Pb2+, respectively. The curves were shifted to more depolarized voltages by Pb2+, indicating that channels were less likely to be inactivated at higher concentrations of Pb2+ at any given potential. The fast (tf) and slow (ts) decay time-constants were both significantly increased by increasing concentrations of Pb2+ (n=10, P<0.05), indicating that Pb2+ increased the decay time-course of TOCs. These effects were concentration-dependent and partly reversible following washing. Ca2+ modulated the TOCs gating and might share same binding site with Pb2+, for which Ca2+ had very low affinity. In summary, the results demonstrated that Pb2+ was a dose- and voltage-dependent, and reversible blocker of TOCs in rat DRG neurons. After Pb2+ application, normal sensory physiology of DRG neurons was affected, and these neurons might display aberrant firing properties that resulted in abnormal sensations. This variation caused by Pb2+ could underlie the toxical modulation of sensory input to the central nervous system. PMID- 11406132 TI - Differential development of Ca2+ dynamics in presynaptic terminal and postsynaptic neuron of the rat auditory synapse. AB - Postnatal development of Ca2+ influx and Ca2+ clearance capacity were investigated in the synapse of medial nucleus of the trapezoid body (MNTB) of rat with fura-2 fluorimetry. In contrast to the presynaptic terminal, Ca2+ dynamics does not basically change in the postsynaptic principal neuron developmentally. This differential development of Ca2+ dynamics between pre- and postsynaptic neurons might be crucial for the organized formation and functional maturation of this synapse. PMID- 11406133 TI - Intraperitoneal injection of antisense peptide nucleic acids targeted to the mu receptor decreases response to morphine and receptor protein levels in rat brain. AB - To determine the effectiveness of peptide nucleic acids (PNAs) in vivo, we designed and synthesized PNAs antisense to the mu receptor, the molecular target of morphine for inducing antinociception. Responsiveness of rats to morphine and the levels of mu receptor expression after treatment was measured. We delivered intraperitoneal injections of antisense PNAs targeted to the mu receptor (AS MOR), mismatch PNAs (AS-MOR MM), antisense PNAs targeted to the neurotensin receptor subtype 1 (AS-NTR1), or saline and then challenged the rats with 5 mg/kg morphine (intraperitonally) or neurotensin directly into the periaqueductal gray region of the brain. To avoid tolerance, separate groups of animals were tested at 24, 48, and 72 h post-PNA treatment. Only animals treated with the AS-MOR showed a reduction in their antinociceptive response to morphine. The lack of effect of morphine on the AS-MOR rats was profound at 24 and 48 h, but animals tested at 72 h were similar to control groups. At 24 h the AS-MOR rats had a significant 55% decrease in the levels of mu receptor in their periaqueductal gray region, while AS-MOR MM rats showed no significant change. Lastly, the AS MOR rats continued to show a normal antinociceptive response to neurotensin. This study, therefore, provides additional support for the use of PNAs to target proteins within brain by systemically administered PNAs. PMID- 11406134 TI - Parasites, parasitology and parasitologists. AB - The science of parasitology is one of the many new disciplines of the twentieth century, as such it is a dynamic and rapidly evolving science which encompasses an increasing number of sub-disciplines and technologies. However, will the fragmentation involved in current methods of scientific enquiry and the competition for funding mean the decline of certain areas of parasitology or perhaps the complete loss of the discipline. This paper attempts to address these questions by considering the development of the discipline of parasitology especially within Australia and by considering the mechanisms of attaining funding for science. Technological change and its impact on parasitology is also considered, and requirements for maintenance of the discipline and its practitioners are suggested. PMID- 11406135 TI - Bancroft--Mackerras oration. Molecular taxonomic, diagnostic and genetic studies of parasitic helminths. PMID- 11406136 TI - Functional analysis of drug resistance in Plasmodium falciparum in the post genomic era. AB - Malaria has plagued humans throughout recorded history and results in the death of over 2 million people per year. The protozoan parasite Plasmodium falciparum causes the most severe form of malaria in humans. Chemotherapy has become one of the major control strategies for this parasite; however, the development of drug resistance to virtually all of the currently available drugs is causing a crisis in the use and deployment of these compounds for prophylaxis and treatment of this disease. The genome sequence of P. falciparum is providing the informational base for the use of whole-genome strategies such as bioinformatics, microarrays and genetic mapping. These approaches, together with the availability of a high resolution genome linkage map consisting of hundreds of microsatellite markers and the advanced technologies of transfection and proteomics, will facilitate an integrated approach to address important biological questions. In this review we will discuss strategies to identify novel genes involved in the molecular mechanisms used by the parasite to circumvent the lethal effect of current chemotherapeutic agents. PMID- 11406137 TI - Post-genomic nematode parasitology. AB - The future direction of post-genomic nematode parasitology should focus on the function of the genes that are defined by large-scale expressed sequence tag sequencing and on broader questions about the genetic basis of parasitism. Functional characterisation will require the application of high throughput technologies that have been developed in other fields, including genome mapping strategies and DNA microarray analysis. These will be greatly aided by the development and application of appropriate model organisms. It is crucial that the field make the transition from a narrow focus on one or a few genes at a time to a focus on whole genomes in order to fully realise the potential of the expressed sequence tag and other genomic projects currently under way. PMID- 11406138 TI - Immune evasion genes from filarial nematodes. AB - Helminth parasites have large genomes (approximately 10(8) bp) which are likely to encode a spectrum of products able to block or divert the host immune response. We have employed three parallel approaches to identify the first generation of 'immune evasion genes' from parasites such as the filarial nematode Brugia malayi. The first strategy is a conventional route to characterise prominent surface or secreted antigens. In this way we have identified a 15-kDa protein, which is located on the surface of both L3 and adult B. malayi, and secreted by these parasites in vitro, as a member of the cystatin (cysteine protease inhibitor) family. This product, Bm-CPI-2, blocks conventional cysteine proteases such as papain, but also the aspariginyl endopeptidase involved in the Class II antigen processing pathway in human B cells. In parallel, we identified the major T cell-stimulating antigen from the microfilarial stage as a serpin (serine protease inhibitor), Bm-SPN-2. Microfilariae secrete this product which blocks two key proteases of the neutrophil, a key mediator of inflammation and innate immunity. The second route involves a priori hypotheses that helminth parasites encode homologues of mammalian cytokines such as TGF-beta which are members of broad, ancient metazoan gene families. We have identified two TGF-beta homologues in B. malayi, and shown that one form (Bm-TGH-2) is both secreted by adult parasites in vitro and able to bind to host TGF-beta receptors. Likewise, B. malayi expresses homologues of mammalian MIF, which are remarkably similar in both structure and function to the host protein, even though amino acid identity is only 28%. Finally, we deployed a third method of selecting critical genes, using an expression-based criterion to select abundant mRNAs taken from key points in parasite life histories. By this means, we have shown that the major transcript present in mosquito-borne infective larvae, Bm-ALT, is a credible vaccine candidate for use against lymphatic filariasis, while a second abundantly expressed gene, Bm-VAL-1, is similar to a likely vaccine antigen being developed against hookworm parasites. PMID- 11406139 TI - Function and assembly of the Leishmania surface coat. AB - Like many trypanosomatids, the cell surface coat of Leishmania spp. is responsible for mediating various host-parasite interactions as well as acting as a dense physical barrier. This confers protection to the parasites in the hostile environments of the sandfly midgut and the macrophage phagolysosome. The major components of the surface coat are tethered to the cell surface via glycosylphosphatidylinositol glycolipids, and the composition of this surface coat is exquisitely regulated during the course of the parasite life-cycle. In this paper, we review what is known about the composition, biosynthesis and function of these glycosylphosphatidylinositol-containing molecules found within the parasite surface coat. PMID- 11406140 TI - Analysis of the sensory responses of parasitic nematodes using electrophysiology. AB - Use of the electrophysiological technique to examine the sensory perception of live, intact nematodes has provided detailed analysis of responses to known concentrations of test chemicals. The use of larger nematodes, such as the animal parasite Syngamus trachea, enabled direct extracellular recordings from individual sensilla; with smaller nematodes, the recording electrode was inserted close to the cephalic region. Extracellular recordings from the cephalic region of second-stage juveniles and males of the potato cyst nematodes, Globodera rostochiensis and Globodera pallida, were obtained after exposure to a variety of semiochemicals, including sex pheromones and certain putative phagostimulatory compounds. The responses of adult females of the animal-parasitic nematode, Brugia pahangi, to some possible host cues, and the inhibition by ivermectin of the response to a known allelochemical were investigated. Exposure to acetylcholine was used to compare the concentration-dependent responses of second stage juveniles of G. rostochiensis and adult females of B. pahangi and the insect-parasitic nematode Leidynema appendiculata. Use of a perfusion system enabled sequential exposure of individual nematodes to different test chemicals or to different concentrations of the same chemical. Incubating second-stage juveniles of G. rostochiensis for 24 h in a mAb showing specificity to amphidial secretions resulted in blocking of the normal response to host root diffusates. The potential of the electrophysiology technique for analysing perturbation of sensory perception is discussed. PMID- 11406141 TI - Environmental, cultural and social changes and their influence on parasite infections. AB - The changes taking place within the societies, cultures and the environments in which we live are massive and complex. By referring to simple epidemiological models it is possible to build an objective framework with which to look at these changes in terms of their likely impact on the epidemiology of parasitic diseases within human communities. These parameters are listed for hosts and both micro- and macroparasites, as are epidemiologically significant cultural, social and environmental variables. Changes in these variables may be either detrimental or beneficial to human health and may, in addition, interact in complex ways. Examples of the complexity of changes which can influence epidemiology are provided for a cultural template of the population living in the north-east of Thailand. PMID- 11406142 TI - The vulnerability of animal and human health to parasites under global change. AB - The term 'global change' is used to encompass all of the significant drivers of environmental change as experienced by hosts, parasites and parasite managers. The term includes changes in climate and climate variability, atmospheric composition, land use and land cover including deforestation and urbanisation, bio-geochemistry, globalisation of trade and transport, the spread of alien species, human health and technology. A subset of land use issues relates to the management of protective technologies in relation to residues in food and the environment and the emergence of resistance. Another is the question of changing biodiversity of both parasites and their associated natural enemies, and the effects on the host--parasite relationship and on parasite management. A framework for studying impacts of global change is proposed and illustrated with field data, and CLIMEX and simulation modelling of the cattle tick Boophilus microplus in Australia. Parasitology suffers from the perception that the key impacts of global change will be driven by changes at lower trophic levels, with parasitic interactions being treated as secondary effects. This is incorrect because the environment mediates host-parasite interactions as much as it affects parasites directly. Parasitologists need to strive for holistic solutions to the management of animal and human health, within a wider context of overall management of those systems, if they are to make a meaningful contribution to global efforts aimed at coping with global change. PMID- 11406143 TI - The future impact of societal and cultural factors on parasitic disease -- some emerging issues. AB - A variety of societal and cultural factors will increase host exposure or susceptibility to infectious agents, particularly parasites. Such factors have already had a major impact on the emergence of infectious diseases and the situation is likely to worsen further as we enter the new millennium. The changes that are enhancing the spread and transmission of parasitic diseases, as well as those which are adversely affecting host responsiveness, are examined with reference to specific parasites. PMID- 11406144 TI - An investigation of the co-evolutionary relationships between onchobothriid tapeworms and their elasmobranch hosts. AB - There is general consensus that the living elasmobranchs comprise a monophyletic taxon. There is evidence that, among tetraphyllidean tapeworms, the approximately 201 hooked species (Onchobothriidae) may also comprise a monophyletic group. Determinations of host specificity are contingent upon correct specific identifications. Since 1960, over 200 new elasmobranch species and over 100 new onchobothriid species have been described. Some confidence can be placed in host and parasite identifications of recent studies, but specific identifications provided in older literature in many cases are suspect. There is some consensus among published works on the phylogenetic relationships among elasmobranchs. Phylogenetic relationships among onchobothriids remain largely unresolved. Elasmobranchs have been poorly sampled for onchobothriids; records exist for approximately 20% of the 911 species and approximately 44% of the 170 elasmobranch genera. Onchobothriids are remarkably host specific, exhibiting essentially oioxenous specificity for their definitive hosts. Multiple onchobothriid species commonly parasitise the same host species; in some cases these are congeners, in other cases these are members of two different onchobothriid genera. There is substantial incongruence between available host and parasite phylogenies. For example, Acanthobothrium is by far the most ubiquitous onchobothriid genus, parasitising almost all orders of elasmobranchs known to host onchobothriids, yet, there is no evidence of major clades of Acanthobothrium corresponding to postulated major subgroupings of elasmobranchs (e.g. Galea and Squalea or sharks and rays). Potamotrygonocestus appears to be among the most basal onchobothriid groups, yet it parasitises one of the most derived elasmobranch groups (the freshwater stingray genus Potamotrygon). It appears that congeners parasitising the same host species are not necessarily each other's closest relatives. At this point the preliminary and limited available data suggest that, at least in this system, strict host specificity is not necessarily indicative of strict co-evolution. This study was extremely limited by the lack of available robust phylogenies for onchobothriids and elasmobranchs. PMID- 11406145 TI - Co-evolutionary relationships between the nematode subfamily Cloacininae and its macropodid marsupial hosts. AB - Morphologically based phylogenies of the cloacinine genera Cyclostrongylus, Macropostrongylus, Pharyngostrongylus, Popovastrongylus, Rugopharynx, Thallostonema, Wallabinema, and Zoniolaimus were constructed and compared with the phylogeny of their respective macropodid hosts. These comparisons show some evidence of co-speciation. However, there was little consistency among trees of different nematode genera, parasite species were scattered amongst hosts and basal parasite taxa were, in some instances, parasitic in hosts belonging to derived clades. A cladistic analysis, using as characters 208 cloacinine nematode species found in 23 species of host, produced a tree largely resembling that of the host tree but with significant differences explainable by host switching among macropodids occupying similar habitat. Nematodes were moderately host specific, but some species occurred in three or more distantly related host species indicating a degree of host switching. The results are more consistent with the hypothesis of a colonisation of macropodid hosts by cloacinine nematodes rather than a prolonged period of co-speciation although alternative interpretations of the data are also considered. PMID- 11406146 TI - The nature and evolution of the association among digeneans, molluscs and fishes. AB - Patterns of association of digenean families and their mollusc and vertebrate hosts are assessed by way of a new database containing information on over 1000 species of digeneans for life-cycles and over 5000 species from fishes. Analysis of the distribution of digenean families in molluscs suggests that the group was associated primitively with gastropods and that infection of polychaetes, bivalves and scaphopods are all the results of host-switching. For the vertebrates, infections of agnathans and chondrichthyans are apparently the result of host-switching from teleosts. For digenean families the ratio of orders of fishes infected to superfamilies of molluscs infected ranges from 0.5 (Mesometridae) to 16 (Bivesiculidae) and has a mean of 5.6. Individual patterns of host association of 13 digenean families and superfamilies are reviewed. Two, Bucephalidae and Sanguinicolidae, are exceptional in infecting a range of first intermediate hosts qualitatively as broad as their range of definitive hosts. No well-studied taxon shows narrower association with vertebrate than with mollusc clades. The range of definitive hosts of digeneans is characteristically defined by eco-physiological similarity rather than phylogenetic relationship. The range of associations of digenean families with mollusc taxa is generally much narrower. These data are considered in the light of ideas about the significance of different forms of host association. If Manter's Second Rule (the longer the association with a host group, the more pronounced the specificity exhibited by the parasite group) is invoked, then the data may suggest that the Digenea first parasitised molluscs before adopting vertebrate hosts. This interpretation is consistent with most previous ideas about the evolution of the Digenea but contrary to current interpretations based on the monophyly of the Neodermata. The basis of Manter's Second Rule is, however, considered too flimsy for this interpretation to be robust. Problems of the inference of the evolution of patterns of parasitism in the Neodermata are discussed and considered so intractable that the truth may be presently unknowable. PMID- 11406147 TI - Analytical approaches to measuring cospeciation of host and parasites: through a glass, darkly. AB - Studies of cophylogenetic associations between hosts and parasites have become increasingly common. Historically, congruence between host and parasite phylogenies has been seen as evidence for cospeciation. Analyses of such coevolutionary relationships, however, are made extremely difficult by the complex interplay of cospeciation, host switching, sorting (extinction), duplication (intrahost speciation) and inertia (lack of parasite speciation) events, all of which may produce incongruence between host and parasite phylogenies. Here we review several methods of analysing cospeciation. We illustrate these methods with an example from a Procellariiformes (seabird) and chewing louse (Halipeurus) association. PMID- 11406149 TI - Coexpression of CD4 and CD8alpha on rat T-cells in whole blood: a sensitive marker for monitoring T-cell immunosuppressive drugs. AB - The aim of this study was to develop a new quantitative method for measuring in vitro the effects of T-cell immunosuppressive drugs by flow cytometry. Rat whole blood samples were stimulated with the T-cell mitogen succinylated concanavalin A in the presence or absence of different drugs. After 3 days, the expression of CD25 and CD8alpha in mitogen-stimulated CD4(+) cells increased 10- to 20-fold as measured by flow cytometry. Drug efficacy and potency was calculated based on dose-response curves of the drug-mediated decrease in CD4(+)/CD8alpha(+)/CD25(+) cells. The expression of CD8alpha in mitogen-stimulated CD4(+) cells was blocked completely by calcineurin inhibitors (cyclosporine A and FK-506), and partially by rapamycin and SDZ-RAD. The IC(50) (50% inhibitory concentration) values obtained were (mean+/-S.E.): 99.5+/-16.6 nM for cyclosporine A, 10.4+/-1.3 nM for FK-506, 1.8+/-0.7 nM for rapamycin, and 6.4+/-1.1 nM for SDZ-RAD. Our results show, for the first time, that CD8alpha, used as an activation antigen, is a sensitive marker for monitoring T-cell immunosuppression. PMID- 11406150 TI - Cyanobacterial stabilized phycobilisomes as fluorochromes for extracellular antigen detection by flow cytometry. AB - Phycobilisomes are cyanobacterial photosynthetic energy transfer complexes partly composed of phycobiliproteins, proteins that are widely used as conjugable fluorochromes for flow cytometry. The brightness and photostability of phycobiliproteins suggest that intact phycobilisomes could constitute even brighter probes for fluorescence-based detection systems. Stabilized phycobilisomes have been isolated and the red-excited, far red-emitting Spirulina platensis-derived complex PBXL-3 was accessed as a fluorochrome for flow cytometric immunodetection of surface antigens on immune cells. Although the large size of intact phycobilisomes initially precluded efficient cell surface labeling, the addition of a PEG spacer arm between PBXL-3 and its conjugated avidin molecule (designated PBXL-3L) reduced the steric hindrance associated with the high molecular weight PBXL complex. PBXL-3L increased the surface labeling surface-to-noise ratio and subsequent sensitivity by several-fold over commonly used red-excited fluorochromes such as APC. Interestingly, low power laser sources (including helium-neon and red diode) were particularly efficient at exciting PBXL-3. PBXL-3 was also compatible in with other fluorochromes for multicolor flow cytometry applications. In summary, PBXL-3 was found to possess superior sensitivity and efficiency for flow cytometric immunodetection, particularly with low power laser sources. PMID- 11406151 TI - A simple method for culturing mouse vascular endothelium. AB - Vascular endothelium is an important site for a wide array of immunological processes such as inflammation, atherosclerosis and allograft rejection. Culture methods of mouse vascular endothelium would provide an important in vitro correlate to immunological murine in vivo models. We describe a simple method to culture mouse vascular endothelium from thoracic aorta. Our cultured cells express typical phenotypic (CD105, CD31, CD106), morphological and ultrastructural (intercellular junctions, Weibel-Palade bodies) markers of vascular endothelium. They also possess functional receptors for uptake and processing of acetylated low-density lipoproteins. The mouse vascular endothelium within our system expresses high levels of MHC class I and MHC class II after activation with IFN-gamma. In addition, these cells express the accessory molecules CD80 and CD54, while they lack constitutive expression of CD86 and CD40, providing them the means to function as antigen presenting cells. Alloreactive CD4(+) and CD8(+) T lymphocytes demonstrate evidence of DNA synthesis after co-culture with activated vascular endothelium indicating their commitment to proliferation. In conclusion, we describe a simple culture system to isolate and grow mouse vascular endothelium, which provides a powerful tool to study biological interactions in vitro. PMID- 11406152 TI - Native and recombinant proteins to analyze auto-antibodies to myeloperoxidase in pauci-immune crescentic glomerulonephritis. AB - The prevalence of Anti-Neutrophil Cytoplasmic Antibodies (ANCA) directed against myeloperoxidase (MPO) in pauci-immune necrotizing crescentic glomerulonephritis (NCGN) is dependent on the assay(s) used. We investigated the frequency of MPO ANCA as detected by different assays for MPO-ANCA in a large cohort of patients with biopsy-proven pauci-immune NCGN. Sera from 121 consecutive untreated patients presenting with pauci-immune NCGN were tested for ANCA directed to proteinase-3 (PR3) at diagnosis. PR3-ANCA negative sera were tested by direct ELISA using recombinant or native MPO and by capture ELISA using two different specific monoclonal antibodies directed to MPO and three different antigenic sources. Sera from 80 relevant disease controls were tested to explore the specificity of the different assays. Thirty-eight out of 121 patients (31%) with pauci-immune NCGN did not have PR3-ANCA. Sufficient amounts of serum from 30 of these 38 PR3-ANCA negative patients were available for further testing. Recombinant and native MPO were recognized by similar numbers of sera in a direct ELISA (recombinant MPO: 93%, native MPO: 93%) and a capture ELISA (recombinant MPO: 77-87%, native MPO: 93%). Sera of patients with PR3-ANCA positive pauci immune NCGN and disease controls were less frequently positive for MPO-ANCA in a capture ELISA (recombinant MPO: 3-7%, native MPO: 6-7%) than in a direct ELISA (recombinant MPO: 25%, native MPO: 13%). Both direct and capture ELISA assays using either native or recombinant MPO are sensitive techniques to detect MPO ANCA in patients with pauci-immune NCGN. A capture ELISA performs better than a direct ELISA because it combines a higher specificity with a comparable sensitivity. Recombinant MPO is a good alternative for native MPO when used as antigen in a capture ELISA, but not when used in a direct ELISA because of lower specificity in this latter assay. PMID- 11406153 TI - A simple and efficient method for the monitoring of antigen-specific T cell responses using peptide pool arrays in a modified ELISpot assay. AB - In this study, we describe a simple and efficient method for both the monitoring of antigen-specific CD4 and CD8 T cell responses as well as the identification of novel CD4 and CD8 T cell epitopes using a modified ELISpot assay and pools of 20mer peptides. We have demonstrated that pools containing as many as 64 20mer peptides may be used to screen for CD4 and CD8 T cell responses to HPV16 L1, E1, and E7 in mice. Using arrays of pools of overlapping 20mer peptides, we have identified novel CD4 and CD8 epitopes in both HPV16L1 and HPV16E1 which are presented in Balb/c mice. We have further shown that the use of 20mer peptides is equivalent to using minimal 9mer epitopes for the stimulation of CD8 T cell responses in our assay. While our experiments are conducted in mice, the use of peptide pool arrays allows for the identification of epitope-specific responses using far fewer cells than is required for testing a panel of overlapping peptides individually, making this strategy particularly useful in clinical settings where immune cells may be limiting. PMID- 11406154 TI - High-throughput generation and engineering of recombinant human antibodies. AB - The first version of the Human Combinatorial Antibody Library (HuCAL) is a single chain Fv-based phage display library (HuCAL-scFv) with 2x10(9) members optimised for high-throughput generation and targeted engineering of human antibodies. 61% of the library genes code for functional scFv as judged by sequencing. We show here that since HuCAL-scFv antibodies are expressed in high levels in Escherichia coli, automated panning and screening in miniaturised settings (96- and 384-well format) have now become feasible. Additionally, the unique modular design of HuCAL-genes and -vectors allows the distinctly facilitated conversion of scFv into Fab, miniantibody and immunoglobulin formats, and the fusion with a variety of effector functions and tags not only convenient for therapeutic applications but also for high-throughput purification and detection. Thus, the HuCAL principle enables the rapid and high-throughput development of human antibodies by optimisation strategies proven useful in classical low molecular weight drug development. We demonstrate in this report that HuCAL is a very convenient source of human antibodies for various applications. PMID- 11406155 TI - Sensitive determination of cell number using the CyQUANT cell proliferation assay. AB - We describe here the development and characterization of the CyQUANT cell proliferation assay, a highly sensitive, fluorescence-based microplate assay for determining numbers of cultured cells. The assay employs CyQUANT GR dye, which produces a large fluorescence enhancement upon binding to cellular nucleic acids that can be measured using standard fluorescein excitation and emission wavelengths. The fluorescence emission of the dye-nucleic acid complexes correlated linearly with cell number over a large range using a wide variety of cell types. Under the recommended assay conditions, standard curves were linear (r(2)>0.995), detecting as few as 10-50 cells and as many as 25,000-50,000 cells with a single dye concentration, depending on cell type. Increasing the dye concentration extended the linear range of the assay to 100,000-250,000 cells. Results of cell proliferation and growth inhibition studies with the assay were similar to those obtained in published studies using other standard assays. CyQUANT assay measurements of serum-stimulated cell proliferation correlated well with measurements made using [3H]-thymidine. Also, the assay was used to analyze cellular DNA or RNA content, with the addition of a nuclease digestion step to the protocol. The assay procedure is simple and convenient, with no wash steps, and is readily amenable to automation. PMID- 11406156 TI - Herpesvirus saimiri transformed T cells and peripheral blood mononuclear cells restimulate identical antigen-specific human T cell clones. AB - Panels of human antigen-specific T cell clones (TCC) have been established by limiting dilution using Herpesvirus saimiri (HVS) subtype C transformed T cells as antigen presenting cells (APC). They showed antigen-specific proliferation when peripheral blood mononuclear cells (PBMC), HVS-transformed T cells and Epstein Barr Virus transformed lymphoblastoid B cell lines (EBV-LCL) were used as APC. All T cell clones were CD4+ and HLA class II restricted. For a detailed analysis, two panels of T cell clones specific for an epitope located in the N terminus of the Merozoite Surface Protein 1 (MSP-1) of Plasmodium falciparum were established from the same founder T cell line using either PBMC or HVS transformed T cells as APC. TCR analysis of the two panels of TCC demonstrated that the same founder cells could be propagated in both culture systems. Furthermore, no difference in the cytokine expression pattern or antigen processing and co-stimulatory requirements was observed between TCC established on PBMC or HVS-transformed T cells. Based on the finding that HVS-transformed T cells can replace PBMC as APC for isolation and propagation of antigen-specific TCC, a protocol was developed and successfully executed, which allows to establish and maintain vaccine-specific T cell clones from 20 ml of blood. This method might be particularly significant in clinical trials of immune intervention strategies. PMID- 11406157 TI - Simultaneous measurement of six cytokines in a single sample of human tears using microparticle-based flow cytometry: allergics vs. non-allergics. AB - Tears play an essential role in maintaining corneal and conjunctival integrity by providing a tightly regulated, optimal extracellular environment critical to its numerous functions, which include anti-microbial defense, wound healing and inflammatory responses such as allergies. Elevated levels of inflammatory cytokines have been reported in tears from various ocular disease states. Characterization of tear cytokines has been limited by the small volume (microliter amounts) attainable. This limitation was addressed with the newly developed Becton Dickinson Cytometric Bead Array (CBA), which combines the principles of the "sandwich" immunoassay with the capability of flow cytometry for simultaneous measurement of the characteristics of multiple particles. This technique allows determination of six human cytokine (IFNgamma, TNFalpha, IL-2, IL-4, IL-5, IL-10) concentrations simultaneously in a single tear sample. Tears were collected from the inferior fornix of non-allergic (n=7) and allergic (n=9) donors. Each tear sample or cytokine standard was incubated with a mixture of capture Ab-bead reagent and detector Ab-phycoerythrin (PE) reagent, and analyzed using flow cytometry. All six cytokines were detectable in both non-allergic and allergic tears. Tears from allergic donors contained significantly less IL-10 (p=0.035), and had significant increases in the ratios of TNFalpha/IFNgamma, IL 5/IFNgamma and IL-5/IL-10 (p=0.0008, 0.0124 and 0.011, respectively). The small volume required (5-10 microl/test) by the Cytometric Bead Array allows measurement of all six cytokines from a single collection of tears. This decreases collection time, minimizing the confounding effect of stimulation on cytokine concentration in tears, as well as allowing calculation of cytokine ratios. PMID- 11406158 TI - Characterization of hybrid CTL epitope delivery systems consisting of the Antennapedia homeodomain peptide vector formulated in liposomes. AB - Peptide carriers, such the homeodomain of Antennapedia molecule (AntpHD), which spontaneously cross cellular membranes, have been exploited to deliver antigenic peptide Cw3 to the major histocompatibility complex (MHC) class-I presentation pathway and to prime cytotoxic T cells (CTL). However, the in vivo use of AntpHD recombinant peptide has been limited because CTLs can only be primed in the presence of sodium dodecyl sulfate (SDS) as adjuvant. In this report, we have exploited liposomes to protect the AntpHD-Cw3 from serum degradation and to facilitate the delivery of the recombinant peptide into the MHC class-I pathway of antigen-presenting cells. We have demonstrated that AntpHD recombinant peptide spontaneously associates with liposomes and this association is stable in vitro. However, exchange studies assessing the transfer of the peptide to model membranes or cells in vitro indicates that approximately 50% of the liposome associated peptide is readily exchangeable. This is consistent with trypsin protection assays, which have shown that approximately 40% of the liposome associated peptide is protected from hydrolysis. Importantly, macrophages and dendritic cells are able to internalize AntpHD recombinant peptide associated with liposomes resulting in efficient delivery of the CTL peptide into the cytosol. These studies have demonstrated that dendritic cells treated with AntpHD Cw3 in liposomes sensitize CTL clones to lyse syngeneic target cells expressing Cw3 epitope. This strategy, which combines liposomes and a peptide vector, provides a new approach for introducing molecules into the MHC class-I antigen presentation pathway of dendritic cells. PMID- 11406159 TI - Chemical engineering of cell penetrating antibodies. AB - Antibodies, being exquisitely specific tools in biology, are routinely used to detect and identify intra-cellular structures. However, current intra-cellular application of antibodies requires that the membrane be rendered leaky, resulting in the death of cells. Here, we present a novel method to allow antibodies to penetrate the cellular membrane of living cells without affecting cell viability. A peptide (MTS, membrane transport sequence) that facilitates transport across membranes has been site-specifically attached to antibodies. MTS-antibodies enter the living cells in culture and can be detected by immunofluorescence and ELISA after extraction. Cellular structures are visualized in living cells using a specific MTS-antibody. Antibodies with membrane penetrating properties can become an important tool for the study of intra-cellular processes in living cells. Furthermore, such membrane penetrating antibodies can be used to selectively stimulate or suppress functions of the cellular machinery. PMID- 11406160 TI - Interaction of the octapeptide angiotensin II with a high-affinity single-chain Fv and with peptides derived from the antibody paratope. AB - The amino-acid sequence of the very high-affinity anti-angiotensin II monoclonal antibody 4D8 was predicted from the nucleotide sequence of the heavy and light chain variable genes. The single-chain variable fragment (scFv) was constructed and expressed in Escherichia coli as a soluble protein and at the surface of the filamentous M13 phage and was compared with the full-length antibody (Ab). The scFv showed the same specificity profile and affinity constant as the intact antibody (5.0x10(10) and 8.0x10(10) M(-1), respectively, by Scatchard analysis). Several peptides from the set of overlapping dodecapeptides covering the variable domains of 4D8 mAb were found to specifically bind biotinylated angiotensin II: peptides from the L1, L2, L3 and H1 regions had the strongest capacity to bind the antigen. PMID- 11406161 TI - Systematic design of mouse Vh gene family-specific oligonucleotides. AB - Kabat's database has often been used to design mouse Vh gene-specific 5' primers. The emphasis was mostly on constructing a universal (degenerate) 5' primer or 5' primer set, which would be able to match every mouse Vh gene. We were interested in finding oligonucleotides that could be used as primers or probes that can discriminate between the different Vh gene families. To this end, Kabat's database was reordered into Vh gene families based on more than 80% homology to prototype Vh gene family sequences. Rules were formulated for adequate annealing of putative primers to their respective genes. Putative primers were derived from the consensus sequences of the Vh gene families. A computer program was designed to systematically screen for the most optimal 5' and 3' Vh gene family-specific primers. This program enabled us to find a set of framework I-specific 5' primers, as well as a set of framework III-specific 3' primers. The found primers were also tested for their use as Vh gene family-specific probes. PMID- 11406162 TI - Selection and characterisation of recombinant single-chain antibodies to the hapten Aflatoxin-B1 from naive recombinant antibody libraries. AB - Selection of antibodies from large repertoire phage display libraries has become a common technique for isolation of specific antibodies to antigens. Many of these libraries are shown to contain antibodies specific to haptens, but only when these haptens are derivatised or conjugated to an immobilising molecule, such as bovine serum albumin (BSA). There has been little demonstration of the suitability of naive recombinant antibody libraries for isolating antibodies that bind low molecular weight haptens in the absence of a carrier molecule and few have addressed the problems associated with selecting antibodies that only recognize the combination of hapten and the carrier molecule. We have panned two phage antibody libraries against AflatoxinB1-BSA and screened single-chain antibody fragments for binding to AflatoxinB1-BSA and Aflatoxin-B1. Many of the antibodies isolated specifically bound AflatoxinB1-BSA, but not soluble Aflatoxin B1 or BSA. Modification of the protocol led to isolation of single-chain fragment variable antibody domain (scFv) antibodies that specifically bound soluble Aflatoxin-B1 with an affinity of 6x10(-9) M. PMID- 11406163 TI - Endothelial cell culture: protocol to obtain and cultivate human umbilical endothelial cells. AB - Endothelial cells play a key role in prominent immunological and pathological processes such as leukocyte trafficking, inflammation, atheroma or cancer cell metastasis. Umbilical veins are probably the most widely used source for human endothelial cells, since they are more easily available than many other vessels, they are free from any pathological process and they are physiologically more relevant than many established cell lines. Here, we describe a standard protocol for preparation, maintenance and quality control of these cells. PMID- 11406164 TI - Panning and selection of proteins using ribosome display. AB - Eukaryotic ribosome complexes can be used as a means to display a library of proteins, and isolate specific binding reagents by screening against target molecules. Here we present, as an example, a method for the display of a library of immunoglobulin variable-like domains (VLDs) for the production of stable mRNA/ribosome/protein complexes. These complexes are produced by the addition of specific in vitro transcriptional promoter elements and translation control sequences to the template DNA. Furthermore, an appropriate spacer (anchor) domain is included for efficient folding of the nascent translated protein, which remains attached to the ribosome complex. Ribosome complexes are panned against hen egg lysozyme-conjugated magnetic beads and genes encoding specific, binding, V-like domains are recovered by RT-PCR and cloned into an Escherichia coli expression vector. PMID- 11406165 TI - Polyreactivity is not an artefact. PMID- 11406166 TI - Recognition of cisplatin adducts by cellular proteins. AB - Cisplatin is a widely used chemotherapeutic agent. It reacts with nucleophilic bases in DNA and forms 1,2-d(ApG), 1,2-d(GpG) and 1,3-d(GpTpG) intrastrand crosslinks, interstrand crosslinks and monofunctional adducts. The presence of these adducts in DNA is through to be responsible for the therapeutic efficacy of cisplatin. The exact signal transduction pathway that leads to cell cycle arrest and cell death following treatment with the drug is not known but cell death is believed to be mediated by the recognition of the adducts by cellular proteins. Here we describe the structural information available for cisplatin and related platinum adducts, the interactions of the adducts with cellular proteins and the implications of these interactions for cell survival. PMID- 11406167 TI - Mechanisms of resistance to cisplatin. AB - The use of cisplatin in cancer chemotherapy is limited by acquired or intrinsic resistance of cells to the drug. Cisplatin enters the cells and its chloride ligands are replaced by water, forming aquated species that react with nucleophilic sites in cellular macromolecules. The presence of the cisplatin adducts in DNA is thought to trigger cell cycle arrest and apoptosis. Knowledge of the mechanism of action of cisplatin has improved our understanding of resistance. Decreased intracellular concentration due to decreased drug uptake, increased reflux or increased inactivation by sulfhydryl molecules such as glutathione can cause resistance to cisplatin. Increased excision of the adducts from DNA by repair pathways or increased lesion bypass can also result in resistance. Finally, altered expression of regulatory proteins involved in signal transduction pathways that control the apoptotic pathway can also affect sensitivity to the drug. An improved understanding of the mechanisms of resistance operative in vivo has identified targets for intervention and may increase the utility of cisplatin for the treatment of cancer. PMID- 11406168 TI - Mutagenicity of TCDD in Big Blue transgenic rats. AB - The compound 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) is a significant environmental contaminant resulting from such industrial processes as pulp and paper production. TCDD is a suspected human carcinogen and its ability to induce cancer in laboratory rodents is well documented. Its mechanism of tumor initiation, however, is not well understood and in vitro mutagenicity studies have yielded inconsistent results. In this study, Big Blue lacI transgenic rats were used to assess the mutagenicity of TCDD in both male and female animals. After 6 weeks of exposure to 2 microg/kg TCDD neither an increase in mutation frequency nor any change in mutation spectrum was observed in either male or female animals. PMID- 11406169 TI - Hamster and rat fetal cells have low spontaneous mutation frequencies and rates. AB - Somatic cells of whole Syrian hamster fetuses (gestation day 13) were isolated and tested by an in vivo/in vitro mutation assay for spontaneous mutation frequencies using independent 6-thioguanine (6-TG), diphtheria toxin (DT), and ouabain mutation selection systems. Optimum conditions were ascertained. For 6-TG mutants, a total of 21 mutants were found in cells from 24 litters on 1993 plates, for an overall mutant frequency of 1.8 x 10(-7) per viable cell with 12 positive litters. In all, 26 litters were tested using DT; 77 mutants were found in 840 plates, yielding an overall mutant frequency of 2.6 x 10(-7), with 20 positive litters. No correlations or familial effects were found among 23 litters tested for both DT and 6-TG. Of 14 litters which were tested for ouabain mutants, 4 were positive, with a total of 5 mutants found on 988 plates, for an overall mutant frequency of 7.6 x 10(-8). For 14 F344 rat fetuses, the overall 6-TG spontaneous mutation frequency was determined to be 1.6 x 10(-7). From the data, estimates of mutation rates were calculated. For mutation to 6-TG resistance the rate was 8.3 x 10(-8), for mutation to DT resistance the rate was 8.1 x 10(-8) and for ouabain, the spontaneous mutation rate was 5.7 x 10(-8). For F344 rat, the spontaneous mutation rate was 1.1 x 10(-7). Induced mutant frequencies after in utero exposure to 1 mmol/kg N-ethyl-N-nitrosourea (ENU) were 311, 135 and 200 times the spontaneous value for 6-TG, DT and ouabain, respectively, for Syrian hamster fetal cells and 125 times the spontaneous 6-TG value for fetal F344 rat cells. Both spontaneous mutation frequencies and underlying spontaneous mutation rates are low, consistent with the view that fetal cells exercise extremely tight control over DNA fidelity. PMID- 11406170 TI - Evidence for gene silencing in Haemophilus influenzae. AB - Evidence for gene silencing of Haemophilus influenzae involved a beta-subunit of RNA polymerase. The gene presumed silenced was rifampin resistance. The evidence that it was silencing, rather than dominance of a rifampin-sensitive marker, was that it took place when the rifampin resistance marker was on both a plasmid and the chromosome, without the presence of a rifampin-sensitive marker, as judged by lack of transformation of a rifampin-resistant cell to rifampin sensitivity by the plasmid. In addition, three compounds that are known to decrease gene silencing in eukaryotes (trichostatin A, sodium butyrate and 5-azacytidine) also decreased the presumed silencing in H. influenzae. Silencing of rifampin resistant Escherichia coli did not take place with the plasmid from H. influenzae. PMID- 11406171 TI - MucA protein affects spontaneous mutation in a localized region of Haemophilus influenzae. AB - A plasmid called pMucA, from a piece of the plasmid pKM101 (Mol. Gen. Genet 167 (1979) 317) cloned in the vector pDM2 (J. Bacteriol. 151 (1982) 1605), caused higher mutation in a local region of Haemophilus influenzae and caused even more mutation there in a strain also containing novC, the latter causing an increase in supercoiling (J. Bacteriol 164 (1985) 525). The novD mutation depressed supercoiling, and also depressed the mutation by pMucA in the local region of the chromosome. Thus, it is clear that supercoiling is an important phenomenon in spontaneous mutation of H. influenzae. The pMucA plasmid caused a number of other phenomena in H. influenzae, induced UV mutation (Proc. Natl. Acad. Sci. USA 82 (1985) 7753), decreased UV sensitivity of transforming DNA, but not cells, and UV induced recombination of mutants of phage HP1c1. The effect of the MucA protein in mutagenesis of H. influenzae we consider to be due to the introduction of some of the E. coli functions from pKM101. We postulate that the localized mutation caused by the MucA plasmid also involved localization of the plasmid or its coded protein in the same area, resulting from binding to a homologous gene, probably rec-1, very close to the localized region. PMID- 11406172 TI - Gyrase mutants affect mutation in a localized region of Haemophilus influenzae. AB - Spontaneous mutation was greatly increased in a localized region of the chromosome of Haemophilus influenzae, but not at other loci, by a nov gene mutation called novC that increased DNA supercoiling. Another nov gene mutation, called novD, decreased spontaneous mutation in the same localized region and depressed DNA supercoiling. Both mutations, which code for the gyrase B subunit, have been cloned, and the cloned versions also altered mutagenesis and supercoiling in a similar fashion as the two mutations on the chromosome, although novC on the plasmid caused somewhat less mutation than on the chromosome. We postulate that the effects of the gyrase B mutations on the chromosome result from their effects on supercoiling because of increased gyrase near its site of production. The fact that the novC on a plasmid does not cause mutagenesis except in the same localized region that is altered by this mutation on the chromosome, is difficult to explain. One possibility is that there is a complex of proteins in this region which is necessary for the effects on supercoiling and thus, also on mutagenesis. PMID- 11406173 TI - Comet assay and early apoptosis. AB - The comet assay is a single cell gel electrophoresis test currently used as a qualitative and quantitative genotoxicity test. However, some of the results from this comet assay and current knowledge on apoptosis lead us to suspect the presence of some false positive results. The aim of this study was to ascertain if apoptotic cells can yield comet images that might distort the interpretation of the results. Using Jurkat cells, that hardly express Fas antigen, and apoptosis induction with anti-Fas antibody, it was possible to show that apoptosis can generate typical comet pictures as soon as the cells enter the apoptosis process. Therefore, comet images cannot be interpreted as a genotoxicity indicator when an apoptosis risk is present. Yopro-1 staining, that is also nearly immediate after apoptosis induction, can be used to balance comet assay results. PMID- 11406174 TI - X-ray induction of microsatellite instability at autosomal loci in human lymphoblastoid WTK1 cells. AB - Many models of carcinogenesis posit that multiple genetic events are required for a normal cell to become cancerous. As the mutation rate of a single gene is in the range of 10(-8) to 10(-5) per cell division, a central question remains, how does a single cell acquire multiple mutations? One hypothesis, originally articulated by Loeb [10], proposed that some mutations may not be isolated events, but are associated with a mutator phenotype that leads to the occurrence of additional mutations elsewhere in the cellular genome. To test this hypothesis, we utilized a human lymphoblastoid cell line (WTK1) that is known to be hypermutable at the autosomal thymidine kinase (TK) locus. We isolated 139 independent clones which were selected for new TK mutations that arose either spontaneously or as the result of a single X-ray exposure of 1.5Gy. These clones were examined for second-site alterations in several microsatellite loci scattered throughout the genome using polymerase chain reaction (PCR) amplification followed by both denaturing gel electrophoresis and single-stranded conformational polymorphism (SSCP) analysis. Of these clones, 21 exhibited second site mutations primarily involving loss of heterozygosity, 17 arose from irradiated cells whereas the remaining four arose from non-irradiated cells. We further examined the 17 clones which exhibited alterations specifically at the D16S265 locus; alterations at this site were associated with an enhanced frequency of mutations at other loci in the same region of chromosome 16q, but were not associated with additional mutations at other sites in the genome. Furthermore, new mutations arose in loci on 16q when these clones were propagated for 6 months in culture. Overall, these results support the hypothesis that radiation can induce a type of genetic instability which may facilitate the occurrence of multiple mutations throughout the genome in a small population of exposed cells. Furthermore, some cells may possess localized regions in the genome which are highly sensitive to the induction of instability. PMID- 11406175 TI - Genetic damage by bifunctional agents in repair-active pre-meiotic stages of Drosophila males. AB - Most of our understanding of germline mutagenesis in Drosophila is based on the DNA repair-inactive, haploid post-meiotic stages. The diploid, repair-active pre meiotic stages are more relevant to the situation encountered in somatic cells. DNA mono-adducts induced by agents like methyl methanesulphonate (MMS) and ethylene oxide (EO) are well repaired in the pre-meiotic cell stages, and these agents show therefore, no or considerable lower mutagenic activity in these stages. In contrast, in this study the two bifunctional nitrogen mustards chlorambucil (CAB) and mechlorethamine (MEC) show significantly elevated mutant frequencies of both post- and pre-meiotic germ cells. Results were similar for the X-chromosomal and the autosomal (2nd) recessive lethal (RL) test. CAB and MEC were also active in stem cells, but in comparison with post-stem cell stages they seem to be better protected. The germ cell specific response in post- and pre meiotic cell stages was for both nitrogen mustards comparable to mutagenic activity patterns observed in the specific locus test in the mouse. It was reported that for diepoxybutane (DEB), another cross-linking agent, the ratio of the RL frequency for the 2nd- and the X-chromosome was increased from 2.1 for post-meiotic stages to 9.5 for pre-meiotic stages. In own experiments aiming to confirm this observation, a high ratio was indeed found. The induction of large deletions by DEB could be the reason for this difference, since such lesions might include both a sex-linked lethal and a vital gene required for the development of spermatocytes into mature sperm. Similar differences were expected for CAB and MEC since they are also inducers of large deletions. But unexpectedly, no differences in 2nd/X RL ratio between post- and pre-meiotic cell stages were found for the nitrogen mustards. Possible causes such as distinct proportions of multi-locus deletions (MLDs), mitotic recombination and the formation of persistent lesions, are discussed. PMID- 11406176 TI - The biological activity of hydrogen peroxide VII. L-Histidine increases incorporation of H(2)O(2) into cells and enhances formation of 8 oxodeoxyguanosine by UV-C plus H(2)O(2) but not by H(2)O(2) alone. AB - L-Histidine (L-His) enhances the clastogenic effects of hydrogen peroxide (H(2)O(2)). We previously suggested the involvement of active transport in the efficient influx of an L-His--H(2)O(2) adduct into cells (Oya-Ohta et al. [1]). In this study, we detected intracellular H(2)O(2) by monitoring formation of 2',7'-dichlorofluorescein (DCF) from its precursor. More fluoroproduct accumulated dose-dependently in cells treated with a mixture of L-His and H(2)O(2) (mixture) than with H(2)O(2) alone. This observation supports our hypothesis that active transport is involved in the enhanced incorporation of H(2)O(2) into cells. Moreover, both mixture and the L-His--H(2)O(2) adduct were less active in the generation of hydroxyl radicals (*OH) upon addition of FeCl(2) than was H(2)O(2) alone in a cell-free system. This result suggests that the Fenton reaction might occur more effectively around the nucleus in cells. An immunohistochemical assay using 8-oxodG-specific monoclonal antibodies did not reveal whether the accumulation of H(2)O(2) generates 8-oxodeoxyguanosine (8 oxodG). No 8-oxodG was evident in cells treated with mixture or with H(2)O(2) alone, or even in cells treated with H(2)O(2) at high doses up to 20 mM and, in some cases, pre-treated with catalase inhibitors. It appears, therefore, that *OH and, specifically, *OH derived from intracellular Fenton reactions, might not play a role in the formation of 8-oxodG. However, exposure to UV-C of cells treated with H(2)O(2) yielded more 8-oxodG in the presence of L-His than in the absence of L-His. Thus, the previously observed enhancing effects of L-His were also noted during the induction of formation of 8-oxodG by UV-C plus H(2)O(2). The formation of 8-oxodG in response to UV-C alone was very limited and, hence, H(2)O(2) seemed to be an effective source of *OH only in the presence of UV-C. It is suggested that the *OH that induces formation of 8-oxodG is not *OH formed via intracellular Fenton reactions but is *OH formed via the dissociation of H(2)O(2) under UV-C. PMID- 11406177 TI - Mutator effects of overproducing DNA polymerase eta (Rad30) and its catalytically inactive variant in yeast. AB - DNA polymerase eta synthesizes DNA in vitro with low fidelity. Based on this, here we report the effects of deletion or increased expression of yeast RAD30 gene, encoding for polymerase eta (Pol eta), on spontaneous mutagenesis in vivo. Deletion of RAD30 did not affect spontaneous mutagenesis. Overproduction of Rad30p was slightly mutagenic in a wild-type yeast strain and moderately mutagenic in strains with inactive 3'-->5'-exonuclease of DNA polymerase epsilon or DNA mismatch repair. These data suggest that excess Rad30p reduces replication fidelity in vivo and that the induced errors may be corrected by exonucleolytic proofreading and DNA mismatch repair. However, the magnitude of mutator effect (only up to 10-fold) suggests that the replication fork is protected from inaccurate synthesis by Pol eta in the absence of DNA damage. Overproduction of catalytically inactive Rad30p was also mutagenic, suggesting that much of the mutator effect results from indirect perturbation of replication rather than from direct misincorporation by Pol eta. Moreover, while excess wild-type Pol eta primarily induced base substitutions in the msh6 and pms1 strains, excess inactive Rad30p induced both base substitutions and frameshifts. This suggests that more than one mutagenic mechanism is operating when RAD30 is overexpressed. PMID- 11406178 TI - Quantitation of heteroplasmy in mitochondrial DNA mutations by primer extension using Vent(R)(exo-) DNA polymerase and RFLP analysis. AB - In this report we describe a simple and rapid protocol for reliable quantitation of mitochondrial DNA (mtDNA) mutations, which is basically a modification of the traditional polymerase chain reaction (PCR)/restriction fragment length polymorphism (RFLP) analysis technique. Up to now, the PCR/RFLP method has been of limited use for the accurate determination of ratios of mutant and wild type molecules, largely owing to the formation of heteroduplex molecules by PCR and incompleteness of restriction digestion. In order to overcome this problem, we have introduced a single-step primer extension reaction using Vent(R)(exo-) DNA polymerase and a fluorescence-labeled primer to the standard assay. The labeled homoduplex molecules are then digested with a restriction endonuclease, and the nucleic acids fractionated on an automated DNA sequencer equipped with GENESCAN analysis software. The amount of mutant mtDNA is readily estimated from fluorescence intensities of the wild-type and mutant mtDNA fragments corrected for incomplete digestion as monitored by a homologous control fragment. The accuracy of the improved protocol was determined by constructing standard curves obtained from defined mixtures of genomic DNA containing homoplasmic wild-type and mutant mtDNA. The expected values were obtained, with an observed correlation coefficient of 0.997 and a typical variability of +/-5% between repeated measurements. Further validation of the protocol is provided by the screening of five patients and unaffected subjects carrying the guanine to adenine transition at the nucleotide 3460 of the mitochondrial genome responsible for the mitochondrial disorder of Leber's hereditary optic neuropathy. PMID- 11406179 TI - DNA breakage due to metronidazole treatment. AB - The mutagenicity of metronidazole [1-(hidroxyethyl)-2-methyl-5-nitroimidazole] (MTZ) has been shown in different prokaryotic systems. However, data on human cells are still contradictory. In this study DNA damage was determined by the single cell gel electrophoresis (SCGE) assay, in lymphocytes from 10 healthy subjects treated with therapeutic doses of this drug. Samples were obtained before treatment, as well as 1 and 15 days after ending treatment. Results showed a significant increase of DNA strand breaks 1 day after ending treatment, although, an inverse correlation between the amount of DNA damage and plasma concentrations of MTZ was obtained. Thus, the observed damage may be induced by some MTZ metabolite rather than by the parent drug. Interestingly, the amount of DNA damage returned to basal levels 15 days after ending treatment, except in two individuals. This persistent damage should be further investigated. PMID- 11406180 TI - Effects of arsenite on p53, p21 and cyclin D expression in normal human fibroblasts -- a possible mechanism for arsenite's comutagenicity. AB - Arsenite, the most likely environmental carcinogenic form of arsenic, is not significantly mutagenic at non-toxic concentrations, but is able to enhance the mutagenicity of other agents. Evidence suggests that this comutagenic effect of arsenite is due to inhibition of DNA repair, but no specific repair enzyme has been found to be sensitive to low (<1 microM) concentrations of arsenite. To determine whether arsenite affects signaling which might alter DNA repair, this study assesses the effect of arsenite on p53-related signal transduction pathways after ionizing radiation. Long-term (14 day) low dose (0.1 microM) arsenite caused a modest increase in p53 expression in WI38 normal human fibroblasts, while only toxic (50 microM) concentrations increased p53 levels after short-term (18 h) exposure. When cells were irradiated (6 Gy), p53 and p21 protein concentrations were increased after 4h, as expected. Both long-term, low dose and short-term, high dose exposure to arsenite greatly suppressed the radiation induced increase in p21 abundance. In addition, long-term, low dose (but not short-term, high dose) exposure to arsenite resulted in increased expression of cyclin D1. These results show that in cells treated with arsenite, p53-dependent increase in p21 expression, normally a block to cell cycle progression after DNA damage, is deficient. At the same time, low (non-toxic) exposure to arsenite enhances positive growth signaling. We suggest that the absence of normal p53 functioning, along with increased positive growth signaling in the presence of DNA damage may result in defective DNA repair and account for the comutagenic effects of arsenite. PMID- 11406181 TI - Mitotic recombination and inactivation in Saccharomyces cerevisiae induced by UV radiation (254 nm) and hyperthermia depend on UV fluence rate. AB - In experiments with wild-type diploid yeast cells of Saccharomyces cerevisiae, the synergistic interaction of ultraviolet (UV) light (wavelength, 254 nm) and heat (45--60 degrees C) was studied both for mutagenic and inactivation effects. Simultaneous hyperthermia and UV light treatments increase the frequency of UV induced mitotic intergenic recombination (crossing-over) and cell inactivation. The enhancing effect was a function of UV light fluence rate. It is concluded that the effect of hyperthermia on low fluence UV or high fluence UV irradiation results in comparable effects on survival and mitotic recombination suggesting similar modulation by hyperthermia of the effects induced by UV at different fluence rates. The interpretation of the data obtained was carried out within the widely accepted point of view considering the synergistic effects as a result of repair ability damage. PMID- 11406182 TI - Low concentration N-methyl-N'-nitro-N-nitrosoguanidine activates DNA polymerase beta expression via cyclic-AMP-protein kinase A-cAMP response element binding protein pathway. AB - Ultraviolet light (UV), ionizing-radiation or alkylating agents are known as carcinogens, mostly because of their ability to damage DNA directly. However, they may also play a diverse role in activating the signal pathways and altering the gene expression. We have shown previously that N-methyl-N'-nitro-N nitrosoguanidine (MNNG) of 0.2 microM can increase the transcription of DNA polymerase-beta gene, which has a cyclic AMP response element (CRE) motif in its promoter. Using the CRE report vector, we show here, such treatment can stimulate the CRE-driven gene expression by approximately 1.5-fold compared with control. Consistent with it, the proportion of ser-133 phosphorylated CRE binding protein (CREB), the related transcription factor was 2.08-fold higher versus control in vero cells after 60 min of MNNG treatment. Although CREB has many putative kinases for its phosphorylation, such as p38 mitogen-activated protein kinase (p38 MAPK), Ca(2+)/calmodulin-dependent protein kinase Pi (CaMK Pi) and protein kinase C (PKC), we found the protein kinase A (PKA) was activated and its activation peaked when cells were treated for 60 min (with arbitrary activity unit of 11.03+/-2.80 and 0.86+/-0.43 in treatment and control, respectively), this phasic character was similar to that of the CREB phosphorylation. We also determined the intracellular cyclic AMP (cAMP) levels and it was found that the cAMP concentration was elevated after 60 min treatment (1.53-fold higher). However, to our surprise, we did not find any accompanying cAMP elevation in cells treated by MNNG for 30 min, in which PKA was activated significantly. These findings, together with other observations, suggest that cAMP-PKA-CREB signaling pathway mediates the low concentration MNNG induced pol-beta expression. In addition to elevated cAMP, there might exist a cAMP-independent PKA activation manner in this course. PMID- 11406183 TI - Differences in sensitivity of murine spermatogonia and somatic cells in vivo to sister-chromatid exchange induction by nitrosoureas. AB - Previously published data indicate that spermatogonia (SPG) are less sensitive to a sister-chromatid exchange (SCE) induction for different mutagens. In an earlier study, we have observed that bromodeoxyuridine (BrdU) substituted murine SPG are less sensitive to SCE induction by gamma ray in cells, than bone marrow (BM) and salivary gland (SG) cells in vivo. This was interpreted to mean that SPG are more efficient in DNA repair or are less prone to SCE induction. That the lower induction of SCE could be due to a reduced accessibility of mutagens to the SPG by virtue of a physiological barrier, was discarded by using gamma radiation. The aim of the present study was to establish whether or not there are differences in SCE induction by nitrosoureas among SPG, SG and BM cells with BrdU substituted or unsubstituted DNA. It was observed that SCE induction by methylnitrosourea (MNU) or by ethylnitrosourea (ENU) in SPG was, respectively, five and two times lower than in SG, and ten and three times lower than in BM. In SPG after BrdU incorporation, there was no increase in efficiency of SCE induction; in fact, there was even a slight decrease by exposure to MNU or ENU. BM and SG cells showed an increased efficiency in SCE induction after BrdU incorporation. This implies that SPG are also less sensitive to SCE induction by nitrosoureas, which cause a different kind of damage from previously assayed mutagens. PMID- 11406184 TI - Bisulfite-induced cytosine deamination rates in E. coli SSB:DNA complexes. AB - E. coli single-stranded binding protein (SSB) has been examined for its ability to modulate bisulfite-induced cytosine deamination rates in single-stranded DNA (ssDNA). We used a lacZ alpha-complementation reversion assay to detect C-->U rates at a single codon in M13mp2 DNA, whether in free ssDNA or in an SSB:ssDNA complex. When incubated at 37 degrees C, the average bisulfite-induced reversion rate constant was four-fold less in SSB:ssDNA complexes than in ssDNA, at a single codon. Across a 250 base pair target and over 23 scorable C-->U sites, the forward rate constant was 4.9-fold less in SSB:ssDNA complexes than in ssDNA alone. After treatment with N-uracil glycosylase, ssDNA incubated with bisulfite had reversion frequencies at the background rate of ssDNA incubated without bisulfite, indicating that virtually all mutations scored were due to C-->U events. The decrease in cytosine deamination rates occurred both in a single codon and over a 250 bp target, indicating that interactions between SSB and ssDNA reduce bisulfite-catalyzed mutations. The structural role of SSB is well recognized in multiple cellular processes; SSB can also function to minimize bisulfite-induced ssDNA mutations. PMID- 11406185 TI - Trans- and cis-DNA adduct concentration in epidermis from mouse and rat skin treated ex vivo with benzo[a]pyrene diol epoxide and its corresponding chlorohydrin. AB - Benzo[a]pyrene diol epoxide, a metabolite of benzo[a]pyrene (BaP), and chlorohydrin, the reaction product of chloride and the epoxide, form in vitro the same trans- and cis-stereoisomeric DNA adducts, but in different proportions. In this study, we asked whether the DNA adduct concentration can be kept the same by applying the appropriate dose of (+/-)-7r,8t-dihydroxy-9t,10t-epoxy-7,8,9,10 tetrahydrobenzo[a]pyrene (anti-BPDE)and (+/-)-7r,8t,9t-trihydroxy-10c-chloro 7,8,9,10-tetrahydrobenzo[a]pyrene (trans-BPDCH) to rodent skin and whether the DNA adducts formed differ only in their trans- and cis-stereoisomerism. Skin from C57Bl6 mice, spontaneous hypertension rats (SHR) and Sprague-Dawley (SD) rats was treated ex vivo immediately after the death of the animals with anti-BPDE and its corresponding bay region chlorohydrin trans-BPDCH and the epidermis was analyzed for DNA adducts 1h after the application. We found that adduct formation at the exocyclic amino groups of deoxyguanosine and deoxyadenosine in epidermal DNA followed a linear dose-response within 6--100 nmol/cm(2) with both chemicals. In order to achieve the same adduct concentration in mouse, spontaneous hypertension rat (SHR), and Sprague-Dawley (SD) rat skin, respectively, a 37-, 23- and 10-fold lower dose of anti-BPDE than of trans-BPDCH had to be applied. The order of 2' deoxyguanosine (dGuo) adduct concentration with anti-BPDE was similar to what has been reported, but the order with trans-BPDCH was (+)-cis-BPDE-N(2)-dGuo adduct>(+)-trans-BPDE-N(2)-dGuo=(-)-trans-BPDE-N(2)-dGuo>(-)-cis-BPDE-N(2)-dGuo in mouse skin. Irrespective of species or strain, a significantly higher proportion of cis-adducts was obtained after treatment with trans-BPDCH than after treatment with anti-BPDE. Therefore, DNA adduct concentration can be kept the same by applying the appropriate dose of anti-BPDE and trans-BPDCH to rodent skin and the DNA adducts formed differ only in their trans- and cis stereoisomerism. PMID- 11406186 TI - Continuing ability of the rodent bone marrow micronucleus assay to act as a predictor of the possible germ cell mutagenicity of chemicals. PMID- 11406187 TI - Oxidative stress induced-neurodegenerative diseases: the need for antioxidants that penetrate the blood brain barrier. AB - Oxidative stress (OS) has been implicated in the pathophysiology of many neurological, particularly neurodegenerative diseases. OS can cause cellular damage and subsequent cell death because the reactive oxygen species (ROS) oxidize vital cellular components such as lipids, proteins, and DNA. Moreover, the brain is exposed throughout life to excitatory amino acids (such as glutamate), whose metabolism produces ROS, thereby promoting excitotoxicity. Antioxidant defense mechanisms include removal of O(2), scavenging of reactive oxygen/nitrogen species or their precursors, inhibition of ROS formation, binding of metal ions needed for the catalysis of ROS generation and up-regulation of endogenous antioxidant defenses. However, since our endogenous antioxidant defenses are not always completely effective, and since exposure to damaging environmental factors is increasing, it seems reasonable to propose that exogenous antioxidants could be very effective in diminishing the cumulative effects of oxidative damage. Antioxidants of widely varying chemical structures have been investigated as potential therapeutic agents. However, the therapeutic use of most of these compounds is limited since they do not cross the blood brain barrier (BBB). Although a few of them have shown limited efficiency in animal models or in small clinical studies, none of the currently available antioxidants have proven efficacious in a large-scale controlled study. Therefore, any novel antioxidant molecules designed as potential neuroprotective treatment in acute or chronic neurological disorders should have the mandatory prerequisite that they can cross the BBB after systemic administration. PMID- 11406188 TI - Novel AMPA receptor potentiators LY392098 and LY404187: effects on recombinant human AMPA receptors in vitro. AB - The present study describes the activity of two novel potent and selective AMPA receptor potentiator molecules LY392098 and LY404187. LY392098 and LY404187 enhance glutamate (100 microM) stimulated ion influx through recombinant homomeric human AMPA receptor ion channels, GluR1-4, with estimated EC(50) values of 1.77 microM (GluR1(i)), 0.22 microM (GluR2(i)), 0.56 microM (GluR2(o)), 1.89 microM (GluR3(i)) and 0.20 microM (GluR4(i)) for LY392098 and EC(50) values of 5.65 microM (GluR1(i)), 0.15 microM (GluR2(i)), 1.44 microM (GluR2(o)), 1.66 microM (GluR3(i)) and 0.21 microM (GluR4(i)) for LY404187. Neither compound affected ion influx in untransfected HEK293 cells or GluR transfected cells in the absence of glutamate. Both compounds were selective for activity at AMPA receptors, with no activity at human recombinant kainate receptors. Electrophysiological recordings demonstrated that glutamate (1 mM)-evoked inward currents in human GluR4 transfected HEK293 cells were potentiated by LY392098 and LY404187 at low concentrations (3-10 nM). In addition, both compounds removed glutamate-dependent desensitization of recombinant GluR4 AMPA receptors. These studies demonstrate that LY392098 and LY404187 allosterically potentiate responses mediated by human AMPA receptor ion channels expressed in HEK 293 cells in vitro. PMID- 11406189 TI - Pharmacological effects of AMPA receptor potentiators LY392098 and LY404187 on rat neuronal AMPA receptors in vitro. AB - The present study describes the pharmacological activity of two novel positive allosteric modulators at AMPA receptors in acutely isolated rat cerebellar Purkinje neurons and cultured rat hippocampal neurons. Currents elicited by application of glutamate (100 microM) to isolated cerebellar Purkinje neurons were potentiated by LY392098, LY404187, cyclothiazide, CX516 and aniracetam. The rank order of potency was LY404187> LY392098> cyclothiazide > CX516> aniracetam. LY392098 displayed a higher maximal efficacy than the other compounds examined. AMPA-activated inward currents in cultured rat hippocampal neurons were potentiated by LY392098, LY404187 and cyclothiazide in a reversible and concentration-dependent manner although considerable heterogeneity in the magnitude of response from cell to cell was observed. LY392098 was ineffective in potentiating AMPA receptor responses when dialyzed via the intracellular solution. The selectivity profiles of the two novel AMPA receptor potentiators were examined. LY392098 or LY404187 had minimal activity on NMDA receptor responses, on voltage-gated calcium channel currents in cultured hippocampal neurons and on GluR5 kainate receptor currents in acutely isolated rat dorsal root ganglion neurons. PMID- 11406190 TI - LY392098, a novel AMPA receptor potentiator: electrophysiological studies in prefrontal cortical neurons. AB - The present experiments investigated the ability of LY392098, a novel positive allosteric modulator of AMPA receptors, to potentiate AMPA receptor-mediated currents of neurons in the prefrontal cortex (PFC). Co-application of LY392098 (0.03-10 microM) with AMPA (5 microM) enhanced current through AMPA receptor/channels in acutely isolated PFC neurons in a concentration-dependent manner. Estimates of the potency (EC(50)) and efficacy for LY392098 yielded an EC(50) value of 1.7+/-0.5 microM and a maximal potentiation of a 31.0+/-4.1-fold increase relative to current evoked by AMPA alone. The potentiation was activity dependent, becoming evident only in the presence of agonist, and time-dependent, continuously developing over prolonged application times. An extracellular site of action was inferred by the absence of potentiation when the compound was applied intracellularly. LY392098 also increased the potency of agonist for the receptor by approximately sevenfold. Selectivity assays showed that the effects of LY392098 were exclusive for AMPA receptors, having no activity at N-methyl-D aspartate (NMDA) receptors in PFC neurons. Extracellular recordings from single PFC neurons in vivo showed that administration of LY392098 (0.001-10 microg/kg, i.v.) enhanced the probability of evoked action potential discharge in response to stimulation of glutamatergic afferents from the ventral subiculum of the hippocampal formation. Spontaneous activity of PFC neurons was also increased. Collectively, these results demonstrate that LY392098 is a highly potent, selective and centrally active positive modulator of native AMPA receptors. PMID- 11406191 TI - Potentiation of responses to AMPA on central neurones by LY392098 and LY404187 in vivo. AB - Enhancement of AMPA receptor mediated synaptic excitation has the potential to aid in the treatment of several psychiatric conditions. To test such claims there is a need to develop more potent compounds than those presently available and to demonstrate that they cross the blood-brain barrier to affect responses at central AMPA receptors. We have now completed in vivo tests with two such compounds, the newly discovered biarylpropylsulfonamides, LY392098 and LY404187, on spinal and hippocampal neurones in anaesthetised rats. In the initial study on spinal neurones, LY392098 (30-1000 microg/kg i.v.) dose-dependently increased responses to iontophoretically administered AMPA but not those to NMDA. Subsequently in a more detailed follow-up study on hippocampal neurones, LY392098 (1-100 microg/kg i.v.) and LY404187 (1-100 microg/kg i.v.) enhanced in a dose dependent manner responses to AMPA. Responses to NMDA were also enhanced but to a less extent. Such enhanced responses to NMDA, but not those to AMPA, were reduced by the NMDA antagonist, ketamine (0.5-1 mg/kg i.v.) whereas the selective AMPA antagonist, LY300168 (GYKI53655; 1 mg/kg i.v.), reduced responses to both NMDA and AMPA. LY392098 also potentiated the synaptic excitation of dentate granule cells following perforant path stimulation. These combined data show that, at doses not dissimilar to those affecting behavioural responses (1-1000 microg/kg; see accompanying papers), the two new drugs cross the blood-brain barrier to affect directly the sensitivity of central AMPA receptors and enhance synaptic excitation in vivo. PMID- 11406192 TI - Binding of an AMPA receptor potentiator ([3H]LY395153) to native and recombinant AMPA receptors. AB - LY395153 is a member of a newly described class of arylpropylsulfonamide AMPA receptor potentiators. Here, we characterize and compare [(3)H]LY395153 binding to native AMPA receptors from rat cerebral cortex and recombinant human GluR4(flip) receptors expressed in HEK293 cells. L-Glutamate and AMPA increased [(3)H]LY395153 binding to both native and recombinant AMPA receptors in a concentration dependent and stereoselective manner; this effect of AMPA receptor agonists reflects an apparent increase in ligand affinity. In the presence of L glutamate (500 microM), [(3)H]LY395153 binding is saturable; the affinity of this radioligand is slightly, albeit statistically significantly higher at human GluR4(flip) (K(d)=55.6+/-5.3nM) than rat cortical receptors (K(d)=110+/-15.1nM). NBQX competitively inhibited L-glutamate-induced increases in [(3)H]LY395153 binding in both native and recombinant receptors, whilst LY303070 (the active isomer of GYKI53655) noncompetitively inhibited this effect in native, but not recombinant receptors. The prototypic AMPA receptor potentiator cyclothiazide competitively inhibited [(3)H]LY395153 binding with a potency (K(i) approximately 7 microM) comparable to EC(50) values reported in electrophysiological studies. In contrast, the structurally unrelated AMPA receptor potentiator CX 516 did not inhibit [(3)H]LY395153 binding at concentrations of up to 600 microM. Further, at concentrations reported to facilitate AMPA receptor desensitization, thiocyanate acts as a competitive inhibitor of [(3)H]LY395153 binding. [(3)H]LY395153 binding was unaffected by a variety of structurally (and mechanistically) diverse compounds tested at a concentration of 10 microM. These data indicate [(3)H]LY395153 is a useful probe for labeling a unique modulatory site on both native and recombinant AMPA receptors. PMID- 11406193 TI - Regulation of BDNF expression in primary neuron culture by LY392098, a novel AMPA receptor potentiator. AB - The effects of a novel AMPA receptor potentiator (LY392098) on the expression of brain-derived neurotrophic factor (BDNF) were examined in primary neuron culture. The addition of either AMPA or LY392098 to cortical neurons elicited a time and concentration dependent increase in mRNA encoding BDNF. Moreover, co-addition of subeffective concentrations of AMPA (1 microM) and LY392098 (1 microM) resulted in dramatic increases in both BDNF mRNA (>25-fold) and protein ( approximately 7 fold) levels, whilst no changes in either NT-3 or NT-4 mRNA were detected. More modest ( approximately 1.5-2.5-fold) elevations in BDNF mRNA and protein expression were also produced by combinations of AMPA and LY392098 in cerebellar granule cell neurons. In contrast, AMPA and LY392098, either alone or in combination, did not elevate BDNF mRNA levels in primary astroglial cultures. Maximum elevations in BDNF mRNA and protein were produced by 6-12h of AMPA receptor activation 1-3h of AMPA receptor activation were required to elevate BDNF mRNA levels. AMPA receptor-mediated increases in BDNF mRNA and protein were abolished by the AMPA antagonist, NBQX, but were unaffected by the NMDA antagonist, MK-801. In cortical neuron cultures, activation of both L-type Ca(+2) channels and mitogen-activated protein (MAP) kinases contribute to AMPA receptor mediated increases in BDNF mRNA. The ability of LY392098 to increase the expression of BDNF in primary neuron culture indicates this and related biarylpropylsulfonamides may be useful in the treatment of neuropsychiatric disorders. PMID- 11406194 TI - Antidepressant-like actions of an AMPA receptor potentiator (LY392098). AB - LY392098 is a member of a novel class of biarylpropylsulfonamides that potentiates AMPA receptor-mediated responses both in vitro and in vivo. In this study, the effects of LY392098 were evaluated in two "behavioral despair" models (the forced swim and tail suspension tests) commonly used to identify clinically useful antidepressants. LY392098 reduced immobility in the forced swim test in both rats and mice, with a minimum effective dose of 0.5 mg/kg (i.p.) in both species. LY392098 (0.1-10 mg/kg, i.p.) did not affect motor activity of rats, indicating that the ability of this compound to reduce immobility in the forced swim test is unrelated to a motor stimulant action. LY392098 also reduced immobility in the tail suspension test in a dose-dependent manner, with a minimum effective dose of 5 mg/kg (i.p). A non-competitive AMPA antagonist (LY300168) blocked the activity of LY392098 in the forced swim test, but did not affect imipramine-induced reductions in immobility. Thus, AMPA receptor activation appears to be required for the antidepressant-like effect of LY392098, but not imipramine. These findings indicate that biarylpropylsulfonamides, exemplified by LY392098, may represent a novel class of antidepressants. PMID- 11406195 TI - Heterogeneity of functional GABA(A) receptors in rat dentate gyrus neurons revealed by a change in response to drugs during the whole-cell current time course. AB - We examined if the drug sensitivity of GABA(A) receptors in dentate gyrus granule neurons changed during the whole-cell current time-course. Effects of drugs on currents evoked immediately (the peak current) upon drug application and currents remaining about two seconds later (semi-plateau current) were compared. The apparent affinity for GABA (EC(50)) of the peak and the semi-plateau current were 14 and 4 microM, respectively. Bicuculline inhibited 50% of the peak and the semi plateau current (IC(50)) at 7 and 36 microM, respectively, while 100 microM was required for full inhibition of the 100 microM GABA-evoked current. Zinc inhibited about 50% of the peak current with an IC(50) value of 94 microM whereas biphasic, but complete inhibition of the semi-plateau current was recorded with IC(50) values of 3 and 558 microM. The decay phase of the 100 microM GABA-evoked current was fitted by a fast (tau(1), 100-300 ms) and a slow (tau(2), 1-2 s) time constants in all cells. The relative current amplitude associated with the fast (A1) and the slow (A2) component varied. The A1 current amplitude appeared more sensitive to bicuculline than the A2 current while the opposite was true for zinc. The results are consistent with heterogenous population of functional GABA(A) receptors in the dentate gyrus granule neurons. PMID- 11406196 TI - Autoradiographic localization of [3H]thiocolchicoside binding sites in the rat brain and spinal cord. AB - Thiocolchicoside is used in humans as a myorelaxant drug with anti-inflammatory and analgesic activity. Recently we established the experimental conditions that allowed the identification of [3H]thiocolchicoside binding sites in synaptic membranes of rat spinal cord and cerebral cortex. The pharmacological characterization of these sites indicated that GABA and several of its agonists and antagonists, as well as strychnine, were able to interact with [3H]thiocolchicoside binding in a dose-dependent manner and with different affinities. In order to gain more insight into the nature and the anatomical distribution of the binding sites labeled by [3H]thiocolchicoside, in the present study we examined the localization of these sites on parasagittal and coronal sections of the rat brain and spinal cord, respectively, using receptor autoradiography. In the spinal cord an intense signal was observed in the gray matter, with the highest density occurring in the superficial layers of the dorsal horns. Strychnine completely displaced [3H]thiocolchicoside binding, whereas GABA only partially removed the radioligand from its binding sites. In the brain, specific binding occurred in several areas and was displaced by both GABA and strychnine. The distribution of [3H]thiocolchicoside binding sites in brain sections, however, did not match that found for [3H]muscimol. Furthermore, cold thiocolchicoside was not able to completely displace [3H]muscimol binding, and showed a different efficacy in the various areas labeled by the radioligand. We conclude that thiocolchicoside may interact with a subpopulation of GABA(A) receptors having low-affinity binding sites for GABA. Furthermore, the observed sensitivity to strychnine in the spinal cord indicates an interaction also with strychnine-sensitive glycine receptors, suggesting that the pharmacological effects of thiocolchicoside may be the result of its interaction with different receptor populations. PMID- 11406197 TI - Multiple structural elements contribute to voltage-dependent facilitation of neuronal alpha 1C (CaV1.2) L-type calcium channels. AB - Voltage- and frequency-dependent facilitation of calcium channel activity has been implicated in a number of key physiological processes. Various mechanisms have been proposed to mediate these regulations, including a switch between channel gating modes, voltage-dependent phosphorylation, and a voltage-dependent deinhibition of G-protein block. Studying such modulation on recombinant Ca channels expressed in oocytes, we previously reported that alpha(1C) L-type calcium channel contrast with non-L type Ca channels by its ability to exhibit facilitation by pre-depolarization (Voltage-dependent facilitation of a neuronal alpha(IC) L-type calcium channel, E. Bourinet et al., EMBO Journal, 1994; 13, 5032-5039). To further analyze this effect, we have investigated the molecular determinants which mediate the differences in voltage-dependent facilitation between "facilitable" alpha(1C) and "non facilitable" alpha(1E) calcium channels. We used a series of chimeras which combine the four transmembrane domains of the two channels. Results show that the four domains of alpha(1C) contribute to facilitation, with domain I being most critical. This domain is required but not sufficient alone to generate facilitation. The minimal requirement to observe the effect is the presence of domain I plus one of the three others. We conclude that similarly to activation gating, voltage-dependent facilitation of alpha(1C) is a complex process which involves multiple structural elements were domains I and III play the major role. PMID- 11406198 TI - Modulation of the hyperpolarisation-activated current, Ih, in rat facial motoneurones in vitro by ZD-7288. AB - ZD-7288 [4-(N-ethyl-N-phenylamino)-1,2-dimethyl-6-(methylamino) pyrimidinium chloride] and Cs(+) have been used to distinguish the currents contributing to inward rectification in neonatal rat facial motoneurones (FMs). ZD-7288 (0.1-10 microM) inhibited a current that reversed at -43.7+/-3.7 mV in artificial cerebrospinal fluid (ACSF) containing 3 mM K(+) (n=9), and displayed the time and voltage dependence of the hyperpolarisation-activated current, I(h). Depolarisation-activated transient (I(K(A))) and sustained outward currents were unaffected by ZD-7288. The IC(50) for block of I(h) by ZD-7288 was around 0.2 microM. Onset of inhibition was slow and no recovery was seen after washing in ZD 7288-free ACSF for up to 4 h. In the presence of ZD-7288, Ba(2+) and Rb(+) blocked an inwardly rectifying potassium (K(+)) current, confirming both the presence of I(K(IR)) and its insensitivity to ZD-7288. Cs(+) rapidly and reversibly blocked both I(h) and I(K(IR)). Inhibition of I(h) by ZD-7288 showed no use dependence, internally applied ZD-7288 also blocked I(h), and tail current analysis indicated inhibition to be voltage-independent. In the presence of internal guanosine 5'-O-(3-thiotriphosphate) (GTP-gamma-S) and after previous exposure to ZD-7288, 5-hydroxytryptamine (5-HT), but not noradrenaline, promoted a recovery of I(h) that was not observed if ZD-7288 was present throughout the recording period. This interaction between ZD-7288 and irreversible 5-HT-receptor activation may be related to the mechanism underlying ZD-7288-mediated block of these channels. PMID- 11406199 TI - Neurotensin excitation of serotonergic neurons in the rat nucleus raphe magnus: ionic and molecular mechanisms. AB - To understand the cellular and molecular mechanisms by which neurotensin (NT) induces an analgesic effect in the nucleus raphe magnus (NRM), whole-cell patch clamp recordings were performed to investigate the electrophysiological effects of NT on acutely dissociated NRM neurons. Two subtypes of neurons, primary serotonergic and secondary non-serotonergic cells, were identified from acutely isolated NRM neurons. During current-clamp recordings, NT depolarized NRM serotonergic neurons and evoked action potentials. Voltage-clamp recordings showed that NT excited serotonergic neurons by enhancing a voltage-insensitive and non-selective cationic conductance. Both SR48692, a selective antagonist of subtype 1 neurotensin receptor (NTR-1), and SR 142948A, a non-selective antagonist of NTR-1 and subtype 2 neurotensin receptor (NTR-2), failed to prevent neurotensin from exciting NRM serotonergic neurons. NT-evoked cationic current was inhibited by the intracellular administration of GDP-beta-S. NT failed to induce cationic currents after dialyzing serotonergic neurons with the anti G(alphaq/11) antibody. Cellular Ca(2+) imaging study using fura-2 showed that NT induced the calcium release from the intracellular store. NT-evoked current was blocked after the internal perfusion of heparin, an IP(3) receptor antagonist, or BAPTA, a fast Ca(2+) chelator. It is concluded that neurotensin enhancement of the cationic conductance of NRM serotonergic neurons is mediated by a novel subtype of neurotensin receptors. The coupling mechanism via G(alphaq/11) proteins is likely to involve the generation of IP(3), and subsequent IP(3) evoked Ca(2+) release from intracellular stores results in activating the non selective cationic conductance. PMID- 11406200 TI - Stimulation of P2 receptors in the ventral tegmental area enhances dopaminergic mechanisms in vivo. AB - It has been shown that endogenous adenosine 5'-triphosphate (ATP) as well as its exogenously applied structural analogue, 2-methylthio ATP (2-MeSATP), facilitate the release of dopamine from axon terminals in the rat nucleus accumbens (NAc) by activating ATP-sensitive P2 receptors. In the present study, reversed microdialysis of 2-MeSATP (10 microM, 100 microM and 1 mM), or its microinjection (0.5, 5.0 and 50 pmol) into the ventral tegmental area (VTA), dose-dependently increased the local extracellular level of dopamine and the locomotion in the open field, respectively. These effects were abolished by the P2-receptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid (PPADS). When applied alone, the antagonist decreased the basal dopamine concentration, indicating that endogenous ATP controls the somatodendritic release of dopamine. Repeated microinjections of 2-MeSATP (5 pmol) once daily for 4 days led to a reproducible locomotor stimulation in the open field. Conditioned locomotion was induced by re-exposure to the novel environment on the seventh day. A challenge with amphetamine (1 mg/kg intraperitoneally) on the eighth day enhanced the locomotor activity in the 2-MeSATP-treated group in the sense of a cross sensitisation, but failed to do so in the control group. Neurons in the VTA were heavily stained with antibodies developed against the P2Y(1) subtype of P2 receptors. Taken together, our data suggest that P2 receptors (probably of the P2Y(1) subtype) are involved in the initiation of somatodendritic dopamine release in the VTA and thereby may have a profound influence on sensitisation and reward-motivated behaviour. PMID- 11406201 TI - Up-regulation of TNF-alpha convertase (TACE/ADAM17) after oxygen-glucose deprivation in rat forebrain slices. AB - Tumour necrosis factor-alpha (TNF-alpha) is a major immunomodulatory and proinflammatory cytokine which is shed in its soluble form by a membrane-anchored zinc protease, identified as a disintegrin and metalloproteinase (ADAM) called TNF-alpha convertase (TACE; ADAM17). The role of this protease in the adult nervous system remains poorly understood. During cerebral ischemia and subsequent reperfusion, expression and release of TNF-alpha have been shown. We have investigated the expression and activity of TACE in an in vitro model of brain ischemia consisting of rat forebrain slices exposed to oxygen-glucose deprivation (OGD). OGD caused the release of TNF-alpha, an effect which was inhibited by a hydroxamate-based metalloprotease inhibitor, BB-3103, with an IC(50) of 0.1 microM, suggesting that TNF-alpha release results selectively from TACE activity. Assay of TACE enzymatic activity on a fluorescein-labelled peptide spanning the cleavage site in pro-TNF-alpha, as well as Western blot and RT-PCR analyses showed that TACE is present in control forebrain and, more interestingly, that TACE expression is increased in OGD-exposed tissue. TACE enzymatic activity from OGD-exposed slices was significantly inhibited by cycloheximide, suggesting that de novo synthesis of TACE contributes to TNF-alpha release after ischaemia. Moreover, it was also inhibited by bisindolylmaleimide I, indicating that TACE activity is regulated by PKC. These findings posed the question of what was its function therein. Among other actions, TNF-alpha has been described to be involved in the expression of inducible nitric oxide synthase (iNOS), a high output NOS isoform associated to cellular damage, but the link between TNF-alpha release after brain ischaemia and iNOS expression in this condition has not been shown. We have now found that iNOS expression in OGD-subjected brain slices is inhibited by BB-3103 at concentrations below 1 microM, indicating that shedding of TNF-alpha by TACE plays a necessary part in the induction of this NOS isoenzyme after OGD. Taken together, these data demonstrate that (1) TACE/ADAM17 activity accounts for the majority of TNF-alpha shedding after OGD in rat forebrain slices, (2) an increase in TACE expression contributes, at least in part, to the rise in TNF-alpha after OGD and (3) iNOS expression in OGD-subjected brain slices results from TACE activity and subsequent increase in TNF-alpha levels. PMID- 11406202 TI - Early cyclosporine concentration as predictor of graft survival in cadaveric renal transplantation. PMID- 11406203 TI - Renal transplant recipients with graft survival longer than 20 years: report on 107 cases. PMID- 11406204 TI - Quality of life after paediatric kidney transplantation: a single-centre experience. PMID- 11406205 TI - Hypertriglyceridemia and low serum HDL cholesterol are common in children after liver transplantation. PMID- 11406206 TI - Paediatric living related donor liver transplantation: experiences from Gothenburg, Sweden. PMID- 11406207 TI - Outcome after liver transplantation for primary sclerosing cholangitis. PMID- 11406208 TI - Results from the familial amyloidotic polyneuropathy world transplant registry. PMID- 11406209 TI - PTLD in liver transplant recipients. PMID- 11406210 TI - Noninvasive cardiac allograft monitoring. PMID- 11406211 TI - CMV infection as a contributory factor for renal allograft injury and loss. PMID- 11406212 TI - Infection of beta-herpesviruses (CMV, HHV-6, HHV-7): role in postrenal transplantation complications. PMID- 11406213 TI - Orthotopic liver transplantation in patients with hepatitis B-related liver disease. PMID- 11406214 TI - Severe clinical course of de novo hepatitis B infection after liver transplantation. PMID- 11406215 TI - Outcome of PTLD in renal and liver allograft recipients. PMID- 11406216 TI - A model for bacterial translocation from small bowel in the rat. PMID- 11406217 TI - Superoxide dismutase in development of obliterative bronchiolitis. PMID- 11406218 TI - Expression of proalpha1(I)- and proalpha1(III)collagen mRNA in chronic allograft rejection in a porcine heterotopic bronchial transplantation model. PMID- 11406219 TI - Expression of platelet-derived growth factor and its receptors in acute rat renal allograft rejection. PMID- 11406220 TI - Serial ultrasensitive CRP measurements may be useful in rejection diagnosis after kidney transplantation. PMID- 11406221 TI - Hyaluronidase reduces intragraft pressure of rejecting tissue. PMID- 11406222 TI - Urinary markers of renal graft rejection. PMID- 11406223 TI - Safety and efficacy of automatic implantable defibrillator used as a bridge to heart transplant. PMID- 11406224 TI - Renal transplantation onto abnormal urinary tract: ileal conduit urinary diversion. PMID- 11406225 TI - Liver transplantation using cavoportal hemitransposition: a possibility in the presence of extensive portal vein thrombosis. PMID- 11406226 TI - Living related liver retransplantation in a 6-month-old child after 60 hours of anhepatic phase following hepatectomy of thrombosed primary liver graft. PMID- 11406227 TI - Iatrogenic lesion at cholecystectomy resulting in liver transplantation. PMID- 11406228 TI - Multivisceral transplantation in Scandinavia: experiences from the first successful five-organ case. PMID- 11406229 TI - Organ donation, allocation, and transplantation in the Nordic countries: Scandiatransplant 1999. PMID- 11406230 TI - Is a 3-day limit for highly urgent liver transplantation for fulminant hepatic failure appropriate, and is the diagnosis in some cases incorrect? PMID- 11406231 TI - Perioperative indicators of poor prognosis after liver transplantation. PMID- 11406232 TI - Early outcome of liver transplantation using donors over 60 years of age. PMID- 11406233 TI - Protective effect of nitric oxide and prostaglandin E(2) in ischemia/reperfusion injury of the liver. PMID- 11406234 TI - Pulmonary effects of caval clamping during liver transplantation without venovenous bypass in acute or chronic liver failure. PMID- 11406235 TI - Effects of L-carnitine-hydrochloride in the cold ischemic preservation of fatty liver grafts. PMID- 11406236 TI - Improvement of postpreservation viability of livers from non-heart-beating donors by fibrinolytic preflush with streptokinase upon graft retrieval. PMID- 11406237 TI - Early function of liver grafts preserved with or without portal perfusion. PMID- 11406238 TI - Elevated Fas-expression and cell death but normal TUNEL detection in experimental liver preservation from non-heart-beating donors. PMID- 11406239 TI - Eicosanoids and delayed graft function in human renal transplantation. PMID- 11406240 TI - Experience with human islet transplantation in Sweden. PMID- 11406241 TI - Experience with 450 adult porcine islet isolation procedures. PMID- 11406242 TI - Procurement of human pancreases for islet isolation-the initiation of a Scandinavian collaborative network. PMID- 11406244 TI - Providing comprehensive palliative care through organ donation. PMID- 11406243 TI - Comparative pharmacokinetic study of neoral versus Sandimmun in Japanese stable renal allograft recipients. PMID- 11406245 TI - Improvement of postischemic hepatic microhemodynamics by an endothelin converting enzyme inhibitor (FR 901533). PMID- 11406246 TI - Reduced doses of hepatitis B immunoglobulin protect against hepatitis B virus infection recurrence after liver transplantation. PMID- 11406247 TI - Surfactant treatment in a non-heart-beating donor rat lung transplantation model. PMID- 11406248 TI - A preliminary evaluation of the discriminative power of the monoethylglycinexylidide formation test after intravenous and oral administration of lidocaine. PMID- 11406249 TI - Abnormal cerebral blood flow findings in transplant patients with posttransplant apraxia of speech. PMID- 11406250 TI - Alloimmune response may be involved in neointimal hyperplasia in cryopreserved aortic allografts. PMID- 11406251 TI - Use of combination cytomegalovirus immune globulin plus ganciclovir for prophylaxis in CMV-seronegative liver transplant recipients of a CMV-seropositive donor organ: a multicenter, open-label study. PMID- 11406252 TI - Insulin sensitivity and beta-cell secretion after liver transplantation in patients with acute liver failure. PMID- 11406253 TI - Hemobilia after mycotic hepatic artery pseudoaneurysm after liver transplantation. PMID- 11406254 TI - Polyomavirus interstitial nephritis in a patient with EBV-negative B-cell posttransplantation lymphoproliferative disorder. PMID- 11406255 TI - Blood and graft eosinophils in acute cellular rejection of liver allografts. PMID- 11406256 TI - FK 506 dose in transplantation: from theory to practice. PMID- 11406257 TI - Clinical and virological course of multiple viral infections after liver transplantation. PMID- 11406258 TI - Sternum transplantation: a novel therapy for aplastic anemia in a rat model. PMID- 11406259 TI - Bronchiolitis obliterans with organizing pneumonia: possible association with human herpesvirus-7 infection after lung transplantation. PMID- 11406260 TI - Porcine small bowel transplantation with FK506 as a single immunosuppressant. PMID- 11406262 TI - An induction versus no-induction protocol in anticalcineurin-based immunosuppression using very low-dose steroids. PMID- 11406263 TI - Is complete avoidance of calcineurin inhibitors or steroids now possible? Importance of patient selection and choice of regimen. PMID- 11406264 TI - Methods and outcomes of calcineurin inhibitor reduction or withdrawal in patients with chronic allograft nephropathy after the first year posttransplantation. PMID- 11406265 TI - Mycophenolate mofetil monotherapy: an optimal, safe, and efficacious immunosuppressive maintenance regimen in kidney transplant patients. PMID- 11406266 TI - Checking out the G(2)/M transition. AB - Tight regulation of cell cycle progression is essential for the maintenance of genomic integrity. The orderly progression from one cell cycle phase to the other is mediated by timed activation of distinct cyclin/cdk complexes. For example, onset of mitosis is regulated by the activation of cyclin B/cdc2 and this event is controlled by several cell cycle checkpoints. Such checkpoints ensure that chromosome segregation does not occur in the case of unreplicated or damaged DNA, or misaligned chromosomes. Recently, new insights into the targets of the DNA damage checkpoint help to unravel more of the complex mechanisms of cell cycle checkpoints. This review focuses on the factors controlling the transition from G(2) phase to mitosis. Also, the pathways contributing to the DNA damage checkpoints in these phases of the cell cycle will be discussed. PMID- 11406267 TI - HMGI/Y proteins: flexible regulators of transcription and chromatin structure. AB - The mammalian HMGI/Y (HMGA) non-histone proteins participate in a wide variety of cellular processes including regulation of inducible gene transcription, integration of retroviruses into chromosomes and the induction of neoplastic transformation and promotion of metastatic progression of cancer cells. Recent advances have contributed greatly to our understanding of how the HMGI/Y proteins participate in the molecular mechanisms underlying these biological events. All members of the HMGI/Y family of 'high mobility group' proteins are characterized by the presence of multiple copies of a conserved DNA-binding peptide motif called the 'AT hook' that preferentially binds to the narrow minor groove of stretches of AT-rich sequence. The mammalian HMGI/Y proteins have little, if any, secondary structure in solution but assume distinct conformations when bound to substrates such as DNA or other proteins. Their intrinsic flexibility allows the HMGI/Y proteins to participate in specific protein-DNA and protein-protein interactions that induce both structural changes in chromatin substrates and the formation of stereospecific complexes called 'enhanceosomes' on the promoter/enhancer regions of genes whose transcription they regulate. The formation of such regulatory complexes is characterized by reciprocal inductions of conformational changes in both the HMGI/Y proteins themselves and in their interacting substrates. It may well be that the inherent flexibility of the HMGI/Y proteins, combined with their ability to undergo reversible disordered-to ordered structural transitions, has been a significant factor in the evolutionary selection of these proteins for their functional role(s) in cells. PMID- 11406268 TI - The mouse beta B1-crystallin promoter: strict regulation of lens fiber cell specificity. AB - Previous studies have shown that the chicken beta B1-crystallin promoter ( 434/+30) contains all of the signals necessary to specifically direct high level expression of heterologous genes to the lens fiber cells of mice. In the present study, the mouse beta B1-crystallin gene was cloned, and its regulation was investigated to further elucidate the mechanisms controlling lens fiber cell specific gene expression. Phylogenetic footprinting analysis of the 5' flanking sequence from the mouse, rat, human and chicken beta B1-crystallin genes identified several known and putative functional cis elements including the PL2 element which is required for lens-specific expression of the chicken beta B1 promoter. Surprisingly, however, all six mouse beta B1-crystallin/CAT constructs tested (-1493/+44, -1493/+30, -870/+30, -250/+30, -135/+30 and -98/+30) were inactive in three different mammalian lens-derived cell lines while only the 870/+30 and -98/+30 constructs were active in chicken primary patched lens epithelial cells. In contrast, the chicken beta B1-crystallin promoter (-434/+30) was transcriptionally active in all lens-derived cells tested. Transgenic mice harboring a mouse beta B1-crystallin -1493/+44 CAT construct did express the transgene specifically in lens fiber cells, however, at lower levels than that previously reported for a chicken -434/+30 CAT construct. These data suggest that, as in other crystallin genes, the regulatory signals controlling lens fiber cell-specific expression are conserved between chicken and mouse. However, the inability of the mouse beta B1-crystallin promoter to function in mammalian lens derived cultured cells implies that this gene has acquired additional cis regulatory elements to ensure lens fiber cell specificity. PMID- 11406269 TI - The replacement H3.3 histone gene in Paracentrotus lividus sea urchin: structure and regulatory elements. AB - We have isolated the Paracentrotus lividus sea urchin H3.3 histone gene and characterized the nucleotide sequences of the gene and its proximal promoter. Band shift experiments showed that two cAMP/PMA responsive elements (CRE/TRE), present in the proximal promoter, bind nuclear factors present in embryos at the blastula and gastrula stages (CRE1) and at the blastula stage (CRE2). The putative H3.3 coding region activating sequences (CRAS) failed to bind nuclear factors while the corresponding elements of the two replication-dependent genes (H3L and late H3) clearly recognized nuclear proteins. These results suggest some role of the CRE/TRE elements but not CRAS elements in the transcriptional regulation of the replication-independent histone genes in invertebrates. PMID- 11406270 TI - Intron-specific RNA binding proteins in the chloroplast of the green alga Chlamydomonas reinhardtii. AB - Mitochondria and chloroplasts both contain group II introns which are believed to be the ancestors of nuclear spliceosomal introns. We used the mitochondrial group II intron rI1 from the green alga Scenedesmus obliquus for biochemical characterization of intron-specific RNA binding proteins. rI1 is correctly spliced from a chloroplast precursor RNA when integrated into the chloroplast genome of Chlamydomonas reinhardtii. Glycerol gradients revealed the sedimentation profile of transcripts containing intron rI1 in native C. reinhardtii extracts and in deproteinized RNA preparations, thus indicating the association of rI1 containing transcripts with high molecular weight ribonucleoprotein complexes in vivo. Furthermore, the specific binding of a 61 kDa protein and a 31 kDa protein with the conserved domain IV was demonstrated using a set of intron derivatives for in vitro RNA binding experiments. We propose that we have biochemically characterized 'general splicing factors', which enable the successful splicing even of mitochondrial introns in chloroplasts. PMID- 11406272 TI - FebA: a gene for eukaryotic translation initiation factor 4E-binding protein (4E BP) in Dictyostelium discoideum. AB - We have identified a gene encoding a eukaryotic initiation factor 4E-binding protein (4E-BP) in the EST database of the Dictyostelium cDNA project. The Dictyostelium 4E-BP, designated febA (four e-binding), showed significant similarity to mammalian 4E-BPs. Northern blot analysis revealed that febA was expressed at a high level in the vegetative growth phase but the level of expression decreased during late development. The gene was shown to be non essential since disruption of the gene had no severe effect; the null mutant proliferated normally and formed normal fruiting bodies. However, strains overexpressing the gene could not be established, suggesting that an excess of FebA protein may have a lethal effect on the cells. PMID- 11406271 TI - Different expression patterns for ubiquitous calpains and Capn3 splice variants in monkey ocular tissues. AB - The purpose of the present investigation was to compare the expression of ubiquitous and tissue-specific calpains in ocular tissues from the Macaca fascicularis monkey. Calpain isoforms in retina and corneal epithelium from adult M. fascicularis monkeys were characterized by RT-PCR, cDNA cloning and sequencing. Calpain isoform activities in ocular tissues were investigated by fractionation on DEAE-HPLC, immunoblotting, and casein zymography. Capn3 splice variants in the ocular tissues from rat, rabbit and monkey were compared after RT PCR. RT-PCR analysis revealed that numerous splice variants of Capn3 were expressed in the epithelium from monkey cornea. The variants contained deletions or insertions in or around the IS1, IS2, and NS regions. The cDNAs for Capn3 variants were highly conserved, yet the expression patterns of the Capn3 isoforms were widely different among the mammalian species. In contrast, the expression patterns of ubiquitous calpains in ocular tissues were conserved among the mammalian species, and similarities between monkey and human cDNAs for Capn1 (mu calpain) and Capn2 (m-calpain) were 98 and 99%, respectively. These results suggested that differences in expression patterns of Capn3 variants might be related to the function of each variant in a particular tissue or species. PMID- 11406273 TI - Molecular cloning and functional characterization of two murine cDNAs which encode Ubc variants involved in DNA repair and mutagenesis. AB - Ubiquitin-conjugating enzyme (Ubc) variants share structural similarity with Ubcs but lack the essential cysteine residue required to form a thioester bond with ubiquitin. Yeast Mms2 is a Ubc variant and plays an important role in error-free DNA postreplication repair to protect cells from killing by DNA damaging agents and mutagenesis. Ironically, one of two known Mms2 homologs, CROC1, has been linked to cell immortalization and tumorigenesis. To further investigate cellular roles played by mammalian Mms2 homologs, we report here the molecular cloning, tissue distribution and functional characterization of two mouse cDNAs encoding mMMS2 and mCROC1. Unlike human CROC1, the mCROC1 gene does not encode two alternative transcripts in most tissues. Instead, nonoverlapping sequences were found in two distinct cDNA clones that together would constitute a full-length open reading frame homologous to CROC1B. Both mMMS2 and the C-terminal mCROC1 core domain are able to complement the yeast mms2 mutant functionally and are able to interact with Ubc13 in a yeast two-hybrid assay, indicating that they are true yeast Mms2 homologs and may play a similar role in DNA postreplication repair. We propose several hypotheses to reconcile the seemingly contradictory observations regarding roles of the two mammalian Mms2 homologs in tumorigenesis and carcinogenesis. PMID- 11406274 TI - The Aspergillus nidulans metE gene is regulated by a second system independent from sulphur metabolite repression. AB - Mutations in the Aspergillus nidulans metE gene lead to requirement for O acetylhomoserine. The gene was cloned by complementation of the metE31 mutation. The coding sequence was found to be interrupted by two introns of 66 and 50 bp, respectively. metE codes for a peptide of 489 amino acids which belongs to the family of homoserine O-acetyltransferases and a well-defined superfamily of alpha/beta hydrolases. Transcription of the metE gene is strongly up-regulated by a severe limitation of methionine, but not of cysteine. This gene is the first sulphur metabolism gene described in A. nidulans which is not regulated by the sulphur metabolite repression system in which cysteine acts as the low-molecular weight effector. PMID- 11406275 TI - Expression of AP-2 alpha in SV40 immortalized human lung fibroblasts is associated with a distinct pattern of cytosine methylation in the AP-2 alpha promoter. AB - Activator protein-2 alpha (AP-2 alpha) is a cell type-specific, developmentally regulated, transcription factor that has been implicated as a critical regulator of gene expression during vertebrate development and carcinogenesis. We found that AP-2 alpha was differentially expressed in the normal human lung fibroblast cell strains WI38, MRC-5 and their respective SV40-transformed cell counterparts WI38-VA, MRC-5VA. Since CpG methylation within genetic regulatory regions has been implicated as a mechanism of gene regulation, we investigated the CpG methylation status of the AP-2 alpha gene promoter in these cells. High resolution mapping of methylated cytosines revealed that differential expression of the AP-2 alpha gene in normal human lung fibroblasts and their SV40 transformed counterparts was associated with distinct patterns of cytosine methylation in the AP-2 alpha promoter just 5' to the transcription initiation site. Site-specific methylation was positively correlated with increased AP-2 alpha gene expression in both transformed cell lines investigated. Interestingly, one of the two major centers of hypermethylation in the transformed cells encompassed the cis-element for the AP-2 repressing transcription factor AP-2rep (KLF12). Finally, a sequence variation in human lung fibroblasts relative to the published sequence revealed a previously unidentified AP-2 binding site at position -528 with respect to the transcription initiation site that overlapped the AP-2rep site. Our results suggest that transcriptional activation of AP-2 alpha in the SV40-transformed cells is mediated, at least in part, by site specific methylation of a negative regulatory cis-element in the AP-2 alpha promoter. PMID- 11406276 TI - Mouse hepatocyte growth factor activator inhibitor type 1 (HAI-1) and type 2 (HAI 2)/placental bikunin genes and their promoters. AB - Hepatocyte growth factor (HGF) activator inhibitor type 1 (HAI-1) and type 2 (HAI 2) were recently discovered as specific inhibitors of HGF activator. Each of them contains two Kunitz-type serine protease inhibitor domains and a transmembrane domain, so that their overall structures are similar to each other. In this study, mouse genes encoding HAI-1 and HAI-2 were cloned by screening of a mouse genomic bacterial artificial chromosome library and by polymerase chain reaction of mouse genomic DNA, respectively. The genes (mHAI-1 and mHAI-2) were defined to consist of 11 and eight exons spanning 11 kbp and 9.5 kbp, respectively. Neither a TATA nor CAAT box was found in 5'-flanking regions of both genes and no apparent homologous portion was observed between mHAI-1 and mHAI-2 promoter regions. Promoter assay of mHAI-1 and human HAI-1 revealed that the potential binding sites of a complex of Egr-1-3 and Sp1, which was well-conserved between human (-42 to -58) and mouse (-44 to -57), might be a key portion of its transcriptional regulation to function as not only house-keeping but also early responsive genes. PMID- 11406277 TI - Two CCAAT boxes in a novel inverted repeat motif are required for Hlx homeobox gene expression. AB - Hlx is a homeobox transcription factor gene required for normal intestinal and hepatic growth in development. We previously found high sequence identity and 17 conserved consensus cis-regulatory/transcription factor binding elements in the mouse and human Hlx 5' regions. A 594 bp sequence in the Hlx 5' region possessing the same activity in driving luciferase expression as larger Hlx 5' sequences had three segments each necessary but not sufficient for luciferase expression in NIH 3T3 cells (which express Hlx). Nine of the conserved putative regulatory elements are positioned within these segments, including two CCAAT boxes on opposite strands within a conserved 44 bp inverted repeat sequence. To test the hypothesis that these elements are required for promoter activity, we compared the reporter expression activity of segments containing mutations of these elements with activity of the parent Hlx promoter sequence. We found that mutation of either CCAAT box or a conserved AP-2 site resulted in a significant decrease in promoter activity. Restoration of the inverted repeat with complementary mutations of both CCAAT boxes did not restore activity. Further, mutation of other portions of the inverted repeat did not affect promoter activity. Mutation of other elements had no effect on promoter activity. PMID- 11406278 TI - Analysis of the mouse Scnn1a promoter in cortical collecting duct cells and in transgenic mice. AB - We have isolated and characterised the promoter of the mouse Scnn1a (alpha ENaC) gene. Using transient transfections of serial deletion mutants into Scnn1a expressing cells, we demonstrate that 1.56 kb of 5' upstream sequence is required for cell-specific expression and corticosteroid-mediated regulation. These 5' sequences are not sufficient to drive expression of a lacZ reporter gene or a rat Scnn1a cDNA in transgenic mice, where they failed to rescue Scnn1a deficiency. PMID- 11406279 TI - Isolation and characterization of the Schizosaccharomyces pombe cDNA encoding the mitochondrial endonuclease(1). AB - We isolated the cDNA of the fission yeast mitochondrial endonuclease SpNUC1, which consists of 322 amino acids and has a significant homology with the budding yeast NUC1 and mammalian endonuclease G. Comparison of the cDNA sequence with the genomic sequence showed that the gene consists of three exons and two introns and spans 1.31 kb. The enzyme localization in mitochondria was demonstrated by expressing the SpNUC1-green fluorescent protein fusion in the yeast. The endonuclease was activated by truncation of the amino-terminal region of the protein, indicating that the enzyme is encoded as an inactive precursor. The active enzyme degraded single-stranded DNA and RNA, the activity being dependent on Mg(2+) (Mn(2+)). PMID- 11406280 TI - Molecular cloning of mouse allantoicase cDNA. AB - The uric acid degradation pathway is progressively lost during vertebrate evolution. In mammals, the end product of this catabolic pathway is allantoin and, therefore, no allantoicase should be present in mouse tissues. Surprisingly, we have found an expressed sequence tag (EST) from mouse testis with high similarity to allantoicase. To characterize this transcript, we have completely sequenced the corresponding EST clone insert and found a 1495 bp long cDNA coding for a 414 amino acid long protein. Identities of mouse versus microorganism allantoicases range from 25 to 30%. Identity reaches 54% when compared to Xenopus allantoicase. Among the tested tissues, only testis possesses the allantoicase transcript. Although no deleterious mutations were found in the coding region, no allantoicase activity could be detected in mouse testis. PMID- 11406281 TI - Novel forms of squamous cell carcinoma antigen transcripts produced by alternative splicing. AB - Squamous cell carcinoma antigen (SCCA) is a member of the ovalbumin serine protease inhibitor family, and the serum level of SCCA is a tumor marker of squamous cell carcinoma. Reverse transcription (RT)-PCR of the squamous cell carcinoma cell line showed the existence of a 156 base shorter transcript compared with that of SCCA1 cDNA. By inverse PCR, we cloned the full length cDNA of this SCCA (SCCA1b). Sequence analysis of the complete 1541 bp SCCA1b cDNA showed that it coded for 338 amino acids and had no typical signal sequence in the NH(2) terminus. The cDNA was expressed in Escherichia coli and the product was detected using Western blotting with antibodies against SCCA. Furthermore, RT PCR of the full coding region of SCCA2 cDNA from cancer tissue showed the existence of a 63 base short transcript (SCCA2b). A comparison of SCCA1b and SCCA2b cDNA with the SCCA1 and SCCA2 genes showed that these messages were derived from each gene by an alternative splicing mechanism. PMID- 11406282 TI - Cloning and characterization of zfBLP1, a Bcl-XL homologue from the zebrafish, Danio rerio. AB - The importance of the Bcl-2 family proteins in normal vertebrate embryogenesis is being recognized; however, their regulatory mechanism is poorly understood. We report here the cloning and characterization of a novel zebrafish Bcl-2 family protein, zfBLP1. The zfBLP1 cDNA is 1942 nucleotides long, encoding a polypeptide of 238 amino acids. The primary sequence of zfBLP1 shares 50% identity to human Bcl-XL, and contains all four conserved BH domains of the Bcl-2 family proteins. Primary sequence analysis identified a consensus ER retention signal at the C terminal end of zfBLP1. Northern blot analysis indicated that there were two major and two minor zfBLP1 mRNA species expressed during embryonic development. Among the two major mRNA species, the short one, approx. 3 kb in size, was expressed throughout embryonic development, while the long one, approx. 7 kb long, was not detectable until the gastrula stage. These results suggest that zfBLP1 is a novel Bcl-2 family protein under complicated regulations, and is likely to play an important role in zebrafish oogenesis and embryogenesis. PMID- 11406283 TI - Cloning and expression pattern of the human NDRG3 gene. AB - We report the cloning and expression pattern of a novel N-myc downstream regulated gene 3 (NDRG3), located on human chromosome 20q11.21-11.23. The NDRG3 cDNA is 2588 base pair in length, encoding a 363 amino acid polypeptide highly related to mouse Ndr3 protein. Northern blot reveals that NDRG3 is highly expressed in testis, prostate and ovary. By in situ hybridization, the NDRG3 mRNA was localized to the outer layers of seminiferous epithelium, indicating that it may play a role in spermatogenesis. PMID- 11406284 TI - Developmental program expression of myosin alkali light chain and skeletal actin genes in the rainbow trout Oncorhynchus mykiss. AB - We have isolated MLC1(F) (tMLC1(F)), MLC3(F) (tMLC3(F)) and skeletal actin cDNAs from the teleost Oncorhynchus mykiss. Sequence analysis indicates that tMLC1(F) and tMLC3(F) are not produced from differentially spliced mRNAs as reported in avians and rodents but are encoded by different genes. Results from RNase protection analysis showed that the corresponding transcripts are expressed in fast skeletal muscles. Whole-mount in situ hybridisation revealed distinct expression patterns of the myosin alkali light chains and skeletal actin genes during skeletal muscle development in the embryo. PMID- 11406285 TI - Cloning and characterization of the major groESL operon from a nitrogen-fixing cyanobacterium Anabaena sp. strain L-31. AB - The major heat-shock-responsive operon groESL has been cloned from the cyanobacterial diazotroph Anabaena. The bicistronic operon harbors an upstream negative regulatory CIRCE element and is transcriptionally activated upon temperature upshift. The deduced amino acid sequence displays strong identity/similarity with other cyanobacterial GroES and GroEL proteins. PMID- 11406286 TI - Cloning, sequencing and expression in the seeds and radicles of two Lupinus albus conglutin gamma genes. AB - Two genes encoding conglutin gamma have been isolated from a Lupinus albus genomic library and sequenced. The expression of conglutin gamma was studied by partial amino acid sequencing of the mature seed protein and by nucleotide sequencing of reverse transcriptase-polymerase chain reaction products from various tissues during the plant life cycle. PMID- 11406287 TI - Effect of LPS administration on the expression of POMC, NPY, galanin, CART and MCH mRNAs in the rat hypothalamus. AB - Anorexia and weight loss are manifestations of inflammation seen both in patients and in experimental animal models such as the lipopolysaccharide (LPS)-treated rat. Using in situ hybridization, the levels of mRNAs encoding proopiomelanocortin (POMC), neuropeptide Y (NPY), galanin, melanin-concentrating hormone (MCH) and cocaine- and amphetamine-regulated transcript (CART) were investigated in the rat hypothalamus after a single intraperitoneal dose (125 microg/kg) of LPS. Four hours after LPS injection the food intake was significantly decreased. POMC and CART mRNA levels were increased in the arcuate nucleus, and MCH, CART and galanin mRNAs were all decreased in the lateral hypothalamic area in LPS-treated rats. Levels of mRNAs for NPY and galanin in the arcuate nucleus, and for MCH and CART in the zona incerta did not change significantly after LPS treatment. These findings support the hypothesis that LPS induced factors mediate signalling to the POMC/CART neurons in the arcuate nucleus which could lead to reduced food intake by decreasing MCH, CART and galanin synthesis in target lateral hypothalamic neurons. PMID- 11406288 TI - Localization of protein Ser/Thr phosphatase 5 in rat brain. AB - Protein phosphatase 5 is a recently discovered Ser/Thr phosphatase that is structurally related to calcineurin and protein phosphatases 1 and 2. Northern blot and in situ hybridization studies have shown that protein phosphatase 5 mRNA is present at high levels in brain and is localized to discrete regions. In the present study, we used immunocytochemistry and immunoblot analyses to examine the regional and subcellular distribution of this enzyme in brain. Our work demonstrates that protein phosphatase 5 is widely expressed throughout brain, but is not uniformly distributed. The most intense staining occurred in neurons of the cerebellum, cerebral cortex, and the supraoptic nucleus of the hypothalamus. Other areas also contained immunoreactive cell bodies, including the globus pallidus, hippocampus, thalamus, lateral preoptic area of the hypothalamus, substantia nigra and other brainstem nuclei. Staining in these cells was observed primarily in perikarya and proximal processes. PMID- 11406289 TI - Cloning of the mouse 5-HT6 serotonin receptor and mutagenesis studies of the third cytoplasmic loop. AB - We have cloned the mouse 5-HT6 serotonin receptor and examined structure-function relationships in the C-terminal end of the third cytoplasmic (CIII) loop, introducing point mutations by site-directed mutagenesis at positions 264 to 268. We examined the ability of 5-HT6 wild type and receptor mutants to activate a cAMP responsive reporter gene when transiently expressed in JEG-3 or COS-7 cells. The wild type 5-HT6 receptor showed strong constitutive activity even when expressed at very low levels and which increased in proportion to the amount of receptor cDNA transfected. Three of the five mutants investigated (K264I, K267A and A268R) showed reduction in constitutive activity compared to wild type. These data suggest that constitutive activity may be important to 5-HT6 receptor activity in vivo and that, unlike some other G-protein coupled receptors, alteration in the BBXXB CIII-loop motif reduces rather than further activates basal activity of the murine 5-HT6 receptor. PMID- 11406290 TI - Beta1 adducin gene expression in DRG is developmentally regulated and is upregulated by glial-derived neurotrophic factor and nerve growth factor. AB - Differential display technique has proven to be effective in identifying differentially regulated genes under a variety of experimental conditions. We identified beta1 adducin as a target in primary rat dorsal root ganglia (DRG) cultures that is upregulated by exposure to nerve growth factor (NGF) and glial derived neurotrophic factor (GDNF). We used real-time reverse-transcription polymerase chain reaction (RT-PCR) for quantitative measurement of beta1 adducin gene expression both in DRG cultures and in vivo. Significant increase in beta1 adducin expression level was observed in DRG cultures treated with either GDNF or NGF, compared to untreated cultures. The expression of beta1 adducin in rat tissues was highest in the brain and high in the cerebellum, superior cervical ganglion and DRG tissues. By contrast, low expression levels of beta1 adducin are detected in sciatic nerve and in non-neural tissues. Our study also showed that expression of beta1 adducin gene is developmentally regulated in rat DRG and trigeminal ganglia, with a peak around P0 and significant attenuation by P21. The level of expression of beta1 adducin in adult rat DRG and trigeminal ganglia may be maintained by the action of neurotrophic factors that are produced in innervated targets like skin and muscle. PMID- 11406291 TI - Quantitative mRNA analysis of five C-terminal splice variants of the human 5-HT4 receptor in the central nervous system by TaqMan real time RT-PCR. AB - 5-HT4 receptors mediate several physiological effects of 5-HT, particularly in the central nervous system (CNS), heart and gut. Recently, several C-terminal splice variants of the human 5-HT4 (h5-HT4) receptor have been described, namely h5-HT4(a), h5-HT4(b), h5-HT4(c), h5-HT4(d) and h5-HT4(g). Previous tissue distribution data suggest some degree of specificity in the mRNA expression patterns of the different h5-HT4 receptor splice variants. However, comparison of the mRNA expression profiles of these splice variants is difficult due to the non quantitative methods used, and in addition, there is very limited data on the expression of each splice variant in human CNS subregions. In the present study we used a single technique, TaqMan real time quantitative RT-PCR, to investigate the mRNA distribution of 5-HT4 receptor C-terminal splice variants in multiple human CNS and peripheral tissues. Using a primer/probe set that amplified all 5 HT4 splice variants (5-HT4pan), the highest CNS expression of 5-HT4 receptor mRNA was observed in basal ganglia, amygdala and hippocampus, consistent with previous studies. h5-HT4(a), h5-HT4(b), h5-HT4(c) and h5-HT4(g) were predominantly expressed in various CNS tissues, compared to most peripheral tissues, but there were differences in expression levels and distribution patterns of each variant. The distribution profile and expression levels observed for the 5-HT4(b) splice variant were virtually identical to that obtained with the 5-HT4pan primer/probe set, whilst the other splice variants were expressed at much lower levels and with different expression patterns obtained with both 5-HT4(b) and 5-HT4pan primer/probe sets. Highest levels of 5-HT4(g) were observed in the hypothalamus and cortex, whilst the 5-HT4(a) variant was highest in the amygdala. 5-HT4(c) expression was highest in the pituitary gland whilst 5-HT4(d) mRNA was only detected in the small intestine at very low levels and not in the CNS. In conclusion, we have shown quantitative differences in the mRNA distribution profiles of the 5-HT4 receptor C-terminal splice variants in human CNS subregions as well as peripheral tissues. In addition, our data suggests that the h5-HT4(b) variant is the most predominant form of the 5-HT4 receptor in humans. PMID- 11406292 TI - Cloning and expression of three K+ channel cDNAs from Xenopus muscle. AB - Embryonic Xenopus muscle cells grown in culture express voltage-gated K+ currents with inactivating and non-inactivating kinetics. Here we report the cloning of three K+ channel cDNAs, designated XKv1.2', XKv1.4 and XKv1.10, from muscle which may underlie these currents. XKv1.2' cDNA appears to be an allelic variant of the XKv1.2 previously cloned from Xenopus. The second cDNA encodes a homologue of Kv1.4 that has not been previously cloned from Xenopus. The predicted XKv1.4 protein shows 73% overall similarity to mouse and chick Kv1.4, but shows significant divergence in the region corresponding to the chain of the inactivating 'ball and chain' domain. The third K+ channel cDNA isolated from Xenopus muscle is a novel Kv1 isoform designated XKv1.10. The predicted protein shares about 70% similarity with other members of the Kv1 subfamily, and about 40% with members of the Kv2, Kv3 and Kv4 subfamilies. XKv1.4 mRNA appears as early as stage 10.5 in whole embryos and is prominent in muscle throughout development from stage 14 to adult. XKv1.2' mRNA is detected by stage 11.5 in whole embryos, but remains at low levels in embryonic skeletal muscle (stages 14 and 21), and is absent from adult muscle. XKv1.10 mRNA is first detected at stage 21 in whole embryos, and is present in muscle from this stage onwards. All three transcripts are present in spinal cord at stage 21. The results support the notion that channels encoded by XKv1.4 contribute to the inactivating K+ current observed in embryonic muscle cells in culture. PMID- 11406293 TI - Regulation of the Y1 neuropeptide Y receptor gene expression in PC12 cells. AB - The Y1 receptor for neuropeptide Y (NPY-Y1) is constitutively expressed in PC12 cells. In this study, we examined the role of nerve growth factor (NGF), pituitary adenylyl cyclase activating polypeptide (PACAP) and dexamethasone on the expression of the gene encoding the rat NPY-Y1 receptor in PC12 cells. A fusion gene (pY1-Luc) was constructed where the reporter enzyme firefly luciferase was placed under the control of 700 bp of the promoter region of the rat NPY-Y1 receptor gene. This promoter region contains recognition consensus sequences for various transcription factors, including one activation protein-1 (AP-1) site, two cyclic AMP responsive element sites, one estrogen receptor element site and four glucocorticoid receptor element sites. NGF increased luciferase activity in a concentration dependent manner. This increase was inhibited by K-252a, a trk A receptor inhibitor, and calphostin C, a PKC inhibitor. PACAP-38 increased luciferase activity in a concentration dependent manner. This activation was inhibited by H-89. Dexamethasone increased transcription of NPY-Y1 gene in PC12 cells. These results indicate that differentiation of PC12 cells into endocrine-like phenotype by dexamethasone and into a neuronal-like phenotype by either NGF or PACAP-38 increases the transcriptional activity of the NPY-Y1 receptor gene in PC12 cells. PMID- 11406294 TI - Co-induction of argininosuccinate synthetase, cationic amino acid transporter-2, and nitric oxide synthase in activated murine microglial cells. AB - Nitric oxide (NO) produced by activated microglia has been implicated in many pathophysiological events in the brain including neurodegenerative diseases. Cellular NO production depends absolutely on the availability of arginine, a substrate of NO synthase (NOS). Murine microglial MG5 cells were treated with bacterial lipopolysaccharide (LPS) and interferon-gamma (IFN-gamma), and expression of inducible NO synthase (iNOS) and arginine-supplying enzymes was investigated by RNA blot analysis. iNOS mRNA was strongly induced after treatment and reached a maximum at 6-12 h. mRNA for argininosuccinate synthetase (AS), a citrulline-arginine recycling enzyme, increased at 6 h and reached a maximum at 12 h. Immunoblot analysis showed that iNOS and AS proteins were also induced. In addition, mRNA encoding the cationic amino acid transporter-2 (CAT-2) was strongly induced shortly after treatment. Induction of mRNAs for iNOS, AS, and CAT-2 by LPS/IFN-gamma was also observed following stimulation of rat primary microglial cells. These results strongly suggest that both arginine transport by CAT-2 and citrulline-arginine recycling are important for high-output production of NO in activated microglial cells. PMID- 11406295 TI - Cell-type non-selective transcription of mouse and human genes encoding neural restrictive silencer factor. AB - Neural-restrictive silencer (NRS) has been identified in at least twenty neuron specific genes, and its nuclear DNA-binding factor, NRSF (also known as RE1 silencing transcription factor (REST)), has been cloned from human and rat, and was shown to repress transcription by recruiting corepressors mSin3 and/or CoREST via its N- and C-terminal domains, leading to chromatin reorganization by mSin3 associated histone deacetylase, HDAC. However, it is largely unknown how NRSF gene expression is regulated. To elucidate the mechanisms for gene expression of NRSF, we isolated the transcriptional unit of the NRSF gene from mouse and human, identified three 5'-non-coding exons in addition to three coding exons, determined transcription start sites, and identified two basal promoter activities in the upstream of the first two non-coding exons. Both promoters functioned equally in neuronal and non-neuronal cells, suggesting that levels of initial transcripts of NRSF gene are similar in neuronal and non-neuronal cells. These results suggest that the level of NRSF gene expression is not determined by transcription per se, and rather is modulated at the post-transcriptional level, e.g. splicing, mRNA stability, and/or post-translational modifications, in a cell specific manner. Consistent with this idea, NRSF protein was apparently present even in neuronal cells and tissues, but was unable to bind to the NRS element, suggesting that NRSF is regulated at least in part post-translationally. PMID- 11406296 TI - Amphetamine administration does not alter protein levels of the GLT-1 and EAAC1 glutamate transporter subtypes in rat midbrain, nucleus accumbens, striatum, or prefrontal cortex. AB - Our laboratory and others have previously shown that glutamate transmission is required for chronic amphetamine-induced neuroadaptations, and that glutamate transmission itself is altered by chronic amphetamine administration. For example, N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4 isoxazole-propionic acid (AMPA) receptor subunit expression are altered in a region- and withdrawal-specific manner. The goal of this study was to determine whether repeated amphetamine administration influences the expression of two glutamate transporter subtypes, GLT-1 and EAAC1. Rats were treated with saline or 5 mg/kg amphetamine for 5 days (chronic saline and amphetamine groups, respectively), or saline for 4 days and 5 mg/kg amphetamine on day 5 (acute amphetamine group), and decapitated 24 h after the last injection. Tissue was dissected from brain regions involved in the psychomotor effects of amphetamine (nucleus accumbens, striatum, prefrontal cortex, ventral tegmental area, and substantia nigra). Levels of GLT-1 and EAAC1 were quantified by Western blotting and normalized to actin levels. We found no significant change in levels of GLT-1 or EAAC1 in response to either acute or chronic amphetamine treatment. These findings suggest that the transporter component of the glutamate system might not play a significant role in the alterations in glutamate transmission observed following repeated amphetamine administration. PMID- 11406297 TI - Extracellular-regulated kinase controls beta-amyloid precursor protein mRNA decay. AB - The precise signaling pathways which contribute to amyloid precursor protein (APP) gene expression remain incompletely characterized. We evaluated the role of protein kinases, calcium and phospholipase C (PLC) in modulating APP mRNA levels. There was a rapid 35-40% reduction in the steady state level of APP mRNA upon stimulation of peripheral blood mononuclear cells (PBMC) with phorbol 12 myristate 13-acetate (PMA), A23187 or ionomycin. However the protein kinase C (PKC), protein kinase A (PKA) or PLC pathways did not mediate these changes in APP mRNA levels. Rather, PMA or ionophore caused a rapid activation of extracellular-regulated kinase (ERK). This effect was independent of PKC and sensitive to U0126. After 4 h of PMA treatment, the remaining APP mRNA became indefinitely stable. We propose a model for the biphasic decay of APP mRNA in which ERK activation by PMA causes sequential upregulation of two APP mRNA binding proteins, nucleolin and hnRNP C. We attribute the initial rapid loss of APP mRNA to the helicase activity associated with nucleolin and later stabilization to hnRNP C binding to the 29 base instability element in the 3'-UTR of APP mRNA. PMID- 11406298 TI - Methamphetamine increases expression of the apoptotic c-myc and L-myc genes in the mouse brain. AB - To clarify the possible mechanisms by which the recreational drug, methamphetamine (METH), induces apoptosis, we investigated its effects on the expression of Myc apoptotic genes. This paper presents the characterization of c myc and L-myc gene transcription in the striatum and the cortex. In addition, the expression of the corresponding proteins was also evaluated. Our observations reveal that c-myc and L-myc were up-regulated by METH at both the mRNA and protein levels. Thus, myc transcription factors might be responsible for some aspects of METH-induced apoptotic processes. PMID- 11406300 TI - Synthesis and characterisation of 4-picrylamino-2,6-dinitrotoluene (PADNT): a new insensitive explosive. AB - A chemoselective reductive method has been achieved for the preparation of 4 picrylamino-2,6-dinitrotoluene (PADNT), a new insensitive energetic material which has been characterised by spectral data and elemental analysis. Some explosive properties of the compound have also been determined and the results indicate that PANDT is quite safe to impact and friction. PMID- 11406301 TI - Analysis of fire and explosion hazards of some hydrocarbon-air mixtures. AB - Hazards caused by leakage of hydrocarbons have long been a problem. In this paper, the critical initiation energy and explosion limits of some hydrocarbon air mixtures have been measured in confined (rectangle shock tube) and unconfined (plastic bag) condition tests. Two dimensionless parameters are suggested to compare the fire and explosion hazards of different hydrocarbons. Additionally, a series of experiments was performed to determine the influence of chemical additives on the fire and explosion hazards of some hydrocarbon-air mixtures in confined (rectangle shock tube) tests. These results relate directly to flammability and reactivity of hydrocarbon air mixtures. Such measurements are very important for hydrocarbon safety. PMID- 11406302 TI - Adsorption of chlorinated hydrocarbon vapors onto soil in the presence of water. AB - Chlorinated solvents partition readily into the vapor-phase in unsaturated soils. Sorption from the vapor-phase affects both transport and recovery. The Brunauer Emmett-Teller (BET) isotherm has been used to model adsorption of chlorinated solvent vapors; however, the BET equation is not accurate above reduced vapor pressures (p/p(sat)) of 0.35. New measurements of chlorinated alkane sorption have been used with a modified BET equation (MBET) to model the adsorption isotherms for five solvents. The MBET equation was found to fit experimental data accurately, especially at high reduced vapor pressures. The capacity of sandy loam soil to adsorb organic vapors decreased with increasing water content, probably due to less soil surface available for sorption. This effect is most evident at reduced vapor pressures greater than 0.5. Adsorption is related to distribution of excess surface energy, the number of molecular layers of water sorbed at the surface, and solute polarity. Results suggest that water films contain areas of high interfacial energy that interact with solute molecules via induced electrostatic forces. PMID- 11406303 TI - Sensitivity of leachate and fine contents on electrical resistivity variations of sandy soils. AB - Laboratory pilot tests were performed to investigate the relationship between electrical resistivity and contaminated soil properties. Three different sandy soils and leachate collected from one of the industrial waste landfill sites in Korea were mixed to simulate contaminated soil conditions. The values of electrical resistivity of the soils were measured using laboratory scaled resistivity cone penetrometer probe. In the experiments, electrical resistivity was observed in terms of water content, unit weight, saturation degree of the soils, and leachate concentration. The experimental results show that the electrical resistivity of the sandy soils depends largely on the water content and electrical properties of pore water rather than unit weight and types of soils. The amount of fines can have significant effect on electrical properties of soils. Direct correlation with contamination in such soils may not be valid here. The results suggest that the electrical resistivity measurement is well suited and applicable for monitoring and delineation of contaminants in the subsurface. PMID- 11406304 TI - Release of Cr(III) from Cr(III) picolinate upon metabolic activation. AB - Hexavalent and trivalent chromium are released into the environment from a number of different industrial activities. It is known that Cr(VI) can be reduced and subsequently complexed by humic acids to produce Cr(III) humic acid complexes in the soil and aquatic environments. The metabolic fate of Cr(III) humic acid complexes and other Cr(III) organic complexes in mammalian systems is unknown. Therefore, Cr(III) picolinate was chosen as a model complex for Cr(III) humic acid complexes and other environmentally relevant Cr(III) complexes. Both human hepatocyte microsomes and primary cultures of chick hepatocytes were used to generate metabolites of Cr(III) picolinate. The results from both of these treatments show that a significant amount of Cr(III) is released (66 and 100%, respectively) and that N-1-methylpicotinamide is the primary organic metabolite from this compound. These data suggest that the populations of humans who are exposed Cr(III) picolinate or other environmentally relevant organic Cr(III) complexes, such as Cr(III) humic acid complexes, are potentially accumulating high levels of Cr(III) intracellularly. This intracellular accumulation of Cr(III) can result in the formation of covalent bonds between Cr(III) and DNA and/or other macromolecules, causing genotoxic effects. These data should be considered when assessing the risk of an area contaminated with chromium. PMID- 11406305 TI - Polycyclic aromatic hydrocarbon (PAH) emissions from a coal-fired pilot FBC system. AB - Due to the extensive amount of data suggesting the hazards of these compounds, 16 polycyclic aromatic hydrocarbons (PAHs) are on the Environmental Protection Agency (EPA) Priority Pollutant List. Emissions of these PAHs in the flue gas from the combustion of four coals were measured during four 1000h combustion runs using the 0.1MW heat-input (MWth) bench-scale fluidized bed combustor (FBC). An on-line sampling system was designed for the 16 PAHs, which consisted of a glass wool filter, condenser, glass fiber filter, Teflon filter, and a Tenax trap. The filters and Tenax were extracted by methylene chloride and hexane, respectively, followed by GC/MS analysis using the selective ion monitoring (SIM) mode. In this project, the effects of operating parameters, limestone addition, chlorine content in the coal, and Ca/S molar ratio on the emissions of PAHs were studied. The results indicated that the emissions of PAHs in an FBC system are primarily dependent on the combustion temperature and excess air ratio. The injection of secondary air with high velocity in the freeboard effectively reduces PAH emissions. The addition of extra limestone can promote the formation of PAHs in the FBC system. Chlorine in the coal can possibly lead to large benzene ring PAH formation during combustion. The total PAH emission increases with an increase in the sulfur content of coal. Incomplete combustion results in PAHs with four or more benzene rings. High efficiency combustion results in PAHs with two or three benzene rings. PMID- 11406306 TI - A review of structure-based biodegradation estimation methods. AB - Biodegradation, being the principal abatement process in the environment, is the most important parameter influencing the toxicity, persistence, and ultimate fate in aquatic and terrestrial ecosystems. Biodegradation of an organic chemical in natural systems may be classified as primary (alteration of molecular integrity), ultimate (complete mineralization; i.e. conversion to inorganic compounds and/or normal metabolic processes), or acceptable (toxicity ameliorated). Most of the biodegradation correlations presented in the literature focus on the characterization of primary or ultimate, aerobic degradation. The US Environmental Protection Agency (USEPA) is charged with determining the risks associated with the thousands of chemicals employed in commerce, an effort that is being facilitated through much research aimed at reliable structure-activity relationships (SAR) to predict biodegradation of chemicals in natural systems. To this end, models are needed to understand the mechanisms of biodegradation, to classify chemicals according to relative biodegradability, and to develop reliable biodegradation estimation methods for new chemicals. Frequently, published correlations associating molecular structure to biodegradation will attempt to quantify the degradability of a limited set of homologous chemicals. These correlations have been dubbed quantitative structure biodegradability relationships (QSBRs). More scarce and valuable to researchers are those models that predict the biodegradability of compounds possessing a wide variety of chemical structures. The latter may use any of several techniques and molecular descriptors to correlate biodegradability: QSBRs, pattern recognition, discriminant analysis, and principle component analysis (PCA), to name several. Generally, models either predict the propensity of a chemical to biodegrade using Boolean-type logic (i.e. whether a chemical will "readily biodegrade" or not), or else they quantify the degree of biodegradation by providing information such as rate constants. Such quantitative predictions of biodegradability come in a diversity of parameters, including half-lives, various biodegradation rates and rates constants, theoretical oxygen demand (ThOD), biological oxygen demand (BOD), and others. In this paper, after describing the advantages and disadvantages of the various biodegradation estimation methods found in the literature, the best models are compared to conclude which provide the greatest utility for determining the biodegradability of chemicals with widely varying structures. The group contribution technique presented by Boethling et al. [Environmen. Sci. Technol. 28 (1994) 459] appears to be the most advantageous for use in broad screening for tendency to biodegrade. The model is simple to use, calculating a probability of biodegrading ranging from 0 (none) to 1 (certain), and has proven to be accurate for a wide range of chemical structures, as established by the large, high-quality data set (BIODEG evaluated biodegradation database, Syracuse Research Corporation, Merrill Lane, Syracuse, NY 13210) used to develop this correlation. The authors, therefore, recommend the method of Boethling et al. [Environ. Sci. Technol. 28 (1994) 459] for the initial screening of chemicals to aid in determining whether additional information is necessary to establish relative biodegradability. For readers with applications requiring more quantitative results, such as biodegradation rate constants, enough model details are presented in this paper to allow the reader to pick a suitable correlation, although the reader is cautioned to consult the original, primary reference for the complete method description, equations, and limitations. PMID- 11406307 TI - Adsorption of hydrazoic acid from aqueous solution by macroreticular resin. AB - Sodium azide is a key component in the automobile air bag. When dissolved in aqueous solution, it reacts rapidly with water to form hydrazoic acid which is a highly toxic chemical and is strongly regulated by government. In the present study, adsorption of hydrazoic acid from aqueous solution by macroreticular resin is investigated. This method can provides a convenient means for dealing with the toxic hydrazoic acid. Experimental tests of batch equilibrium adsorption and continuous column adsorption of hydrazoic acid were conducted and the test results were employed to establish adsorption isotherm and to evaluate the column adsorption efficiency. The test results revealed that the multilayer adsorption isotherms, like the modified Langmuir or Jossens model, are needed to adequately describe the hydrazoic acid adsorption equilibrium between the liquid and solid (resin) phases. In the column adsorption process, a theoretical model was adopted for representing the hydrazoic acid change in the aqueous solution exiting the column and the verified theoretical model significantly facilitates prediction of adsorption breakthroughs and column design. Regeneration of exhausted resin was investigated. Solution of 10% (w/w) NaCl was found to be a very efficient regenerant. PMID- 11406308 TI - A combined treatment approach using Fenton's reagent and zero valent iron for the removal of arsenic from drinking water. AB - Studies on the development of an arsenic remediation approach using Fenton's reagent (H2O2 and Fe(II)) followed by passage through zero valent iron is reported. The efficiency of the process was investigated under various operating conditions. Potable municipal water and ground water samples spiked with arsenic(III) and (V) were used in the investigations. The arsenic content was determined by ICP-QMS. A HPLC-ICPMS procedure was used for the speciation and determination of both As(III) and (V) in the processed samples, to study the effectiveness of the oxidation step and the subsequent removal of the arsenic. The optimisation studies indicate that addition of 100 microl of H2O2 and 100 mg of Fe(II) (as ferrous ammonium sulphate) per litre of water for initial treatment followed by passing through zero valent iron, after a reaction time of 10 min, is capable of removing arsenic to lower than the US Environmental Protection Agency (EPA) guideline value of 10 microg/l, from a starting concentration of 2 mg/l of As(III). Using these suggested amounts, several experiments were carried out at different concentrations of As(III). Residual hydrogen peroxide in the processed samples can be eliminated by subsequent chlorination, making the water, thus, processed, suitable for drinking purposes. This approach is simple and cost effective for use at community levels. PMID- 11406309 TI - The absorption kinetics of NO in NaClO2/NaOH solutions. AB - A combined wet scrubbing SOx/NOx removal system is an advanced air pollution control process. In this process, the mechanism of NO removal is relatively unknown. Consequently, absorption of NO by alkaline solutions of NaClO2 was studied to clarify the reaction kinetics. The experiments were carried out mainly at temperature 50 degrees C which is the common operating temperature for wet scrubbers. The assumption that the absorption occurred under the fast pseudo-mth reaction regime was verified in the experiments. The absorption rate of NO into NaClO2 solutions was found to be proportional to P(2)NO,0 and [NaClO2]0(2). The addition of NaOH to solutions of NaClO2 decreased the absorption rate of NO. The absorption rate of NO at 25 degrees C is lower than at 50 degrees C. In the study, the absorption rate of NO did not change with changing gas flow rates. PMID- 11406310 TI - Microorganism selection and biosurfactant production in a continuously and periodically operated bioslurry reactor. AB - A continuous-flow reactor (CSTR) and a soil slurry-sequencing batch reactor (SS SBR) were maintained in 8l vessels for 180 days to treat a soil contaminated with diesel fuel (DF). Concentrations of Candida tropicalis, Brevibacterium casei, Flavobacterium aquatile, Pseudomonas aeruginosa, and Pseudomonas fluorescens were determined using fatty acid methyl ester (FAME) analysis. DF removal (biological and volatile) and biosurfactant concentrations were measured. The SS-SBR encouraged the growth of biosurfactant-producing species relative to the CSTR. Counts of biosurfactant-producing species (C. tropicalis, P. aeruginosa, P. fluorescens) relative to total microbial counts were 88% in the SS-SBR and 23% in the CSTR. Biosurfactants were produced in the SS-SBR to levels of nearly 70 times the critical micelle concentration (CMC) early in the cycle, but were completely degraded by the end of each cycle. No biosurfactant production was observed in the CSTR. DF biodegradation rates were over 40% greater and DF stripping was over five times lower in the SS-SBR than the CSTR. However, considerable foaming occurred in the SS-SBR. Reversing the mode of operation in the reactors on day 80 caused a complete reversal in microbial consortia and reactor performance by day 120. These results show that bioslurry reactor operation can be manipulated to control overall reactor performance. PMID- 11406311 TI - A procedure for production of adapted bacteria to degrade chlorinated aromatics. AB - Production of biomass adapted to the degradation of a mixture of chlorobenzene (CB) and 1,2-dichlorobenzene (DCB) was investigated in a batch culture with substrates supplied by pulses. CB and o-DCB concentrations which gave the best adapted biomass productivity were determined and found to be 150 and 30 microl l( 1), respectively. The biomass productivity was 51 mg l(-1) h(-1). The biomass yield was 0.38 g of biomass dry weight per gram of substrate. The pulses of 200 microl CB and 40 microl o-DCB, were inhibitory to the bacterial culture. Among the metabolites, muconic acid was found in large quantities in the medium and in the cells. At a time between two pulses of 60 min, adding 150 microl CB and 30 microl o-DCB per each pulse, 7.6g l(-1) of biomass was obtained. The produced biomass served as an inoculum for the biotrickling filter which treated industrial waste gases contaminated by CBs. The method of adapted biomass production was described using CBs, but the degradation of any other toxic volatile pollutant can be improved using this technique. PMID- 11406312 TI - Assessment of electrokinetic removal of heavy metals from soils by sequential extraction analysis. AB - Electrokinetic remediation of metal-contaminated soils is strongly affected by soil-type and chemical species of contaminants. This paper investigates the speciation and extent of migration of heavy metals in soils during electrokinetic remediation. Laboratory electrokinetic experiments were conducted using two diverse soils, kaolin and glacial till, contaminated with chromium as either Cr(III) or Cr(VI). Initial total chromium concentrations were maintained at 1000mg/kg. In addition, Ni(II) and Cd(II) were used in concentrations of 500 and 250mg/kg, respectively. The contaminated soils were subjected to a voltage gradient of 1 VDC/cm for over 200h. The extent of migration of contaminants after the electric potential application was determined. Sequential extractions were performed on the contaminated soils before and after electrokinetic treatment to provide an understanding of the distribution of the contaminants in the soils. The initial speciation of contaminants was found to depend on the soil composition as well as the type and amounts of different contaminants present. When the initial form of chromium was Cr(III), exchangeable and soluble fractions of Cr, Ni, and Cd ranged from 10 to 65% in kaolin; however, these fractions ranged from 0 to 4% in glacial till. When the initial form of chromium was Cr(VI), the exchangeable and soluble fractions of Cr, Ni and Cd ranged from 66 to 80% in kaolin. In glacial till, however, the exchangeable and soluble fraction for Cr was 38% and Ni and Cd fractions were 2 and 10%, respectively. The remainder of the contaminants existed as the complex and precipitate fractions. During electrokinetic remediation, Cr(VI) migrated towards the anode, whereas Cr(III), Ni(II) and Cd(II) migrated towards the cathode. The speciation of contaminants after electrokinetic treatment showed that significant change in exchangeable and soluble fractions occurred. In kaolin, exchangeable and soluble Cr(III), Ni(II), and Cd(II) decreased near the anode and increased near the cathode, whereas exchangeable and soluble Cr(VI) decreased near the cathode and increased near the anode. In glacial till, exchangeable and soluble Cr(III), Ni(II), and Cd(II) were low even before electrokinetic treatment and no significant changes were observed after the electrokinetic treatment. However, significant exchangeable and soluble Cr(VI) that was present in glacial till prior to electrokinetic treatment decreased to non-detectable levels near the cathode and increased significantly near the anode. In both kaolin and glacial till, low migration rates occurred as a result of contaminants existing as immobile complexes and precipitates. The overall contaminant removal efficiency was very low (less than 20%) in all tests. PMID- 11406313 TI - Removal of contaminant metals from fine grained soils, using agglomeration, chloride solutions and pile leaching techniques. AB - A leaching process based on the use of a HCl-CaCl2 solution, with total chloride concentration 4M, was investigated for the removal of contaminant metals from fine acidic soils. The possibility to apply this treatment on piles constructed on-site was also examined as a low cost treatment option. The soil sample used in the study was fine in texture, i.e. clay loam, acidic (pH 5.6), and contaminated mainly with Pb, up to 16000mg Pb/kg dry soil, due to past mining activities. The experimental work comprised all the treatment stages, including agglomeration of fine soil particles to increase the permeability of soil, leaching of the agglomerated soil in a laboratory column, removal of metals from the leachate, regeneration and recycling of the leaching solution and final washing of the treated soil. The initial agglomeration treatment resulted in the formation of coarse aggregates and the percolation of leaching solution through the soil column was maintained at high levels, i.e.75ml/cm(2) per day, during the whole treatment. A low amount of HCl acid was required for the removal contaminants from this particular soil, i.e. 0.44mol HCl/kg soil, due to the absence of acid consuming minerals. The extractions achieved were 94% for Pb, 78% for Zn and more than 70% for Cd. The co-dissolution of soil matrix was very limited, with a total weight loss about 3.5%. The final pH of the soil after washing was found to be 5.15, i.e. slightly lower compared to the initial pH of the soil. The results of this study indicate that chloride leaching, in combination with agglomeration and pile leaching techniques, can be a cost effective option for the removal of metal contaminants from fine acidic soils. PMID- 11406314 TI - Abnormal patterns of microtubule-associated protein-2 (MAP-2) immunolabeling in neuronal nuclei and Lewy bodies in Parkinson's disease substantia nigra brain tissues. AB - Parkinson's disease (PD) is a neurodegenerative disorder associated with the appearance of cytoplasmic Lewy bodies (LBs) in dopaminergic neurons of the substantia nigra and the progressive loss of these neurons. Cytoskeleton alterations and associated impairments of neuronal transport may contribute to neuronal death. Microtubule-associated protein-2 (MAP-2), a cytoskeleton protein is localized primarily in neuronal dendrites and is known to stabilize microtubule assembly and mediate their interactions with other neuronal cell components. To determine if alterations in MAP-2 morphology are present in PD neurons, we used single and double immunohistochemical and immunofluorescent techniques to characterize MAP-2 in PD neuronal tissues. We report abnormal MAP-2 immunolabeling in some neurons of the substantia nigra of PD brain tissues, which were not observed in the normal, age-matched, control brain tissues. Furthermore, MAP-2 was co-localized with alpha-synuclein and ubiquitin in cytoplasmic LBs of neurons. Surprisingly, MAP-2 was also found to form fibrous aggregates and crystal-like structures within neuronal nuclei. These PD-associated alterations in MAP-2 morphology and distribution suggest that impaired neuronal transport may contribute to the progression of neuronal loss in the brains of PD patients. PMID- 11406315 TI - Co-distribution of Fos- and mu opioid receptor immunoreactivity within the rat septopreoptic area and hypothalamus during acute glucose deprivation: effects of the mu receptor antagonist CTOP. AB - Mu opioid receptors occur throughout the brain, but central sites where ligand neuromodulatory effects occur during glucopenia have not been identified. The present studies investigated whether septal, preoptic, and hypothalamic neurons that express immunoreactivity for this receptor are transcriptionally activated in response to the glucose antimetabolite, 2-deoxy-D-glucose (2DG), and if intracerebroventricular (icv) administration of the selective mu receptor antagonist, CTOP, modifies this functional response to glucose substrate imbalance. Neurons labeled for mu receptor-immunoreactivity (-ir) were observed in the lateral septal nucleus (LS), medial septum (MS), anterior division of the stria terminalis (BSTa), median preoptic nucleus (MEPO), medial preoptic nucleus (MPN), parastrial nucleus (PS), anterior hypothalamic periventricular nucleus (PVa), and lateral hypothalamic area (LPO). 2DG injection (400 mg/kg i.p.) resulted in co-labeling of mu receptor-positive neurons in the LS, MS, BSTa, MEPO, PVa, and LPO for nuclear Fos-ir. Icv delivery of CTOP decreased mean numbers of co-labeled neurons in the LS, MS, BSTa, and MEPO. These results provide evidence for transactivational effects of glucopenia on mu opioid receptor-expressing neurons within the septum, preoptic area, and hypothalamus, and suggest that the functional status of these receptors within discrete septopreoptic sites may be critical for maximal glucoprivic induction of the Fos stimulus-transcription cascade within local cells. These results thus support the view that the neural loci described above may serve as substrates for regulatory effects of mu opioid receptor ligands on central compensatory activities during acute glucose deprivation. PMID- 11406316 TI - Decreased expression of the NADH:ubiquinone oxidoreductase (complex I) subunit 4 in 1-methyl-4-phenylpyridinium -treated human neuroblastoma SH-SY5Y cells. AB - Oxidative stress and mitochondrial dysfunction have been implicated in Parkinson's disease (PD) pathology. NADH:ubiquinone oxidoreductase (complex I) (EC 1.6.99.3) enzyme activity is aberrant in both PD and 1-methyl-4 phenylpyridinium (MPP(+)) models of PD. Reverse transcription polymerase chain reaction of RNA isolated from MPP(+)-treated human neuroblastoma SH-SY5Y cells identified changes in steady-state mRNA levels of the mitochondrial transcript for subunit 4 of complex I (ND4). Expression of ND4 decreased to nearly 50% after 72 h of MPP(+) (1 mM) exposure. The expression of other mitochondrial transcripts did not change significantly under the same conditions. Pre-incubation of cells with the free-radical spin-trap, N-tert-butyl-alpha-(2-sulfophenyl)-nitrone prior to MPP(+) exposure, prevented decreases in cell viability and ND4 expression. This suggests that functional defects in complex I enzyme activity in PD and MPP(+) toxicity may result from changes in steady-state mRNA levels and that free radicals may be important in this process. PMID- 11406317 TI - The role of N-methyl-D-aspartate receptors in synaptic plasticity of rat visual cortex in vitro: effect of sensory experience. AB - We examined the role of N-methyl-D-aspartate (NMDA) receptors in synaptic plasticity of visual cortex of light (LR) and dark (DR) reared adult rats in vitro. Layer IV stimulation resulted in field potentials in layer II/III, consisting of two excitatory postsynaptic potentials (EPSP) called EPSP1 and EPSP2. Tetanic stimulation induced long-term potentiation (LTP) in EPSP2 of both LR and DR visual cortices. NMDA receptor antagonist D, L-2-amino-5-phosphono valeric acid (AP5) completely blocked the LTP of EPSP2 in DR visual cortex while it reduced slightly the extent of LTP of EPSP2 in LR ones. Another NMDA receptor antagonist ketamine blocked potentiation of EPSP1 as well as EPSP2 in both groups. Our findings demonstrate that dependency of LTP on NMDA receptors and/or sensitivity of these receptors to the antagonists are different in LR and DR animals. PMID- 11406318 TI - Increased expression of the transcription factor E2F1 during dopamine-evoked, caspase-3-mediated apoptosis in rat cortical neurons. AB - The transcription factor E2F1 mRNA and protein levels increased in rat cortical neurons in response to dopamine (DA)- or 6-hydroxydopamine (OHDA)-evoked apoptosis. Increased E2F1 protein was detected in the nucleus of neurons by double fluorescent immunocytochemistry using antibodies to E2F1 and NeuN. DA and 6-OHDA induced caspase-3-mediated apoptosis of cortical neurons which was attenuated by the addition of antioxidants or caspase-3 inhibitors to the cultures. Antioxidants prevented DA-evoked neuronal apoptosis, and also attenuated the increase in E2F1 expression. These findings suggest that increased expression of the transcription factor E2F1 may serve as a death signal during DA evoked neuronal apoptosis. PMID- 11406319 TI - Temporal change of hippocampal enkephalin and dynorphin mRNA following trimethyltin intoxication in rats: effect of anticonvulsant. AB - Trimethyltin (TMT), an organic metal, has been known to induce behavioral abnormalities including seizures and aggression. We administered TMT to rats, then, behavioral changes as well as the changes of dynorphin and Met-enkephalin mRNA were observed with or without phenobarbital treatment in order to reveal the role of neuropeptides in seizure-generating mechanisms. Met-enkephalin mRNA was significantly increased at the 2nd to 6th day after TMT administration when seizure was frequently observed. Meanwhile, dynorphin mRNA was decreased significantly from the 2nd day to 16th day during aggression score remained high. Phenobarbital abolished not only seizures and aggression, but also the changes of neuropeptide expressions. These results suggest that the changes of dynorphin mRNA are more strongly associated with aggression than seizures, while Met enkephalin changes correlate more with seizures. PMID- 11406320 TI - Short-term administration of fluoxetine and venlafaxine decreases corticosteroid receptor mRNA expression in the rat hippocampus. AB - Chronic treatment with antidepressant drugs (2 weeks or longer) increases corticosteroid receptor mRNA expression in the hippocampus and reduces hypothalamic-pituitary-adrenal axis activity in parallel with improving mood and neuroendocrine function. Earlier effects are less well documented. We examined the effects of short term (9 days) treatment with fluoxetine (10 mg/kg) and venlafaxine (10 mg/kg) on hippocampal mineralocorticoid receptor (MR) and glucocorticoid receptor (GR) mRNA expression and spatial memory in adult rats. In situ hybridization histochemistry showed that the antidepressants decreased MR mRNA expression in all hippocampal subregions (e.g. 45% decrease in CA1 with venlafaxine, P<0.001), while GR mRNA expression was selectively reduced in the CA3 subregion. There was a trend for decreased plasma corticosterone levels following fluoxetine (50% fall, P=0.07) and venlafaxine (30% fall, P=0.18) but neither antidepressants affected spatial memory in the watermaze. Thus antidepressants can have complex and opposing actions on hippocampal corticosteroid receptor expression depending on the duration of treatment. PMID- 11406321 TI - In vitro expression and regulation of ciliary neurotrophic factor and its alpha receptor subunit in neonatal rat olfactory ensheathing cells. AB - During development and in the adult, ciliary neurotrophic factor (CNTF) is expressed at high levels in the olfactory system. In the present study, we asked whether neonatal rat olfactory ensheathing cells (OECs) express CNTF- and CNTF receptoralpha (CNTFRalpha)-mRNA in vitro and studied the regulation of both transcripts in response to growth factor and forskolin (FSK) treatment. We show here that OECs in vitro express CNTF and CNTFRalpha-mRNA under control conditions. Administration of FSK increased the expression of CNTFRalpha while lowering the levels of CNTF. Contrary to fibroblast growth factor-2, CNTF did not stimulate the proliferation of OECs. The observation that OECs express both the ligand and part of its receptor complex may indicate that CNTF exerts paracrine and/or autocrine effects in vivo, which apparently do not include the regulation of cell division. PMID- 11406322 TI - Idebenone in patients with Friedreich ataxia. AB - Friedreich ataxia (FA), the most common form of degenerative ataxia, is thought to be caused by respiratory deficiency due to mitochondrial iron accumulation and oxidative stress. Idebenone, a free-radical scavenger, protects mitochondrial function in in vitro models of FA. In a placebo-controlled crossover trial we studied the effect of idebenone on respiratory function in nine ambulant FA patients. (31)P magnetic resonance spectroscopy demonstrated mitochondrial impairment in vivo in skeletal muscle of all FA patients, but no recovery with idebenone. No effects were seen in clinical scores. Echocardiography did not confirm a preliminary study reporting improvement of FA-associated cardiomyopathy with idebenone. PMID- 11406323 TI - Two mutations in the IV/S4-S5 segment of the human skeletal muscle Na+ channel disrupt fast and enhance slow inactivation. AB - Fast and slow inactivation (FI, SI) of the voltage-gated Na+ channel are two kinetically distinct and structurally dissociated processes. The voltage sensor IV/S4 and the intracellular IV/S4-S5 loop have been shown to play an important role in FI mediating the coupling between activation and inactivation. Two mutations in IV/S4-S5 of the human muscle Na+ channel, L1482C/A, disrupt FI by inducing a persistent Na+ current, shifting steady-state inactivation in the depolarizing direction and accelerating its recovery. These effects were more pronounced for L1482A. In contrast, SI of L1482C/A channels was enhanced showing a more complete SI and a 3-fold slowing of its recovery. Effects on SI were more pronounced for L1482C. The results indicate an important role of the IV/S4-S5 loop not only in FI but also in SI of the Na+ channel. PMID- 11406324 TI - Differential effects of cocaine on local cerebral glucose utilization in the mouse and in the rat. AB - The aim of the study was to determine whether cocaine causes a different pattern of functional changes in the rat as compared to the mouse. The [(14)C]2 deoxyglucose method, for measuring local cerebral metabolic rates for glucose, was carried out in Sprague-Dawley rats and in two strains of mice, C57BL/6 and DBA/2, following a single intravenous administration of cocaine. Cocaine, according to previous reports, increased glucose utilization in the nucleus accumbens of the rat, while the drug decreased metabolic rates in most of brain areas of both strains of mice. The post-hoc analysis, however, suggests that the pattern of metabolic changes differ in the two strains. In particular, the effect on the shell of the nucleus accumbens was present in the C57, but not in the DBA mice. As the C57 mice are more likely than DBA to initiate cocaine self administration, the effect on the nucleus accumbens support the role of the mesolimbic pathway in mediating the motivational properties of psychostimulants. PMID- 11406325 TI - Disruption of actin filaments causes redistribution of ryanodine receptor Ca2+ channels in honeybee photoreceptor cells. AB - Ca2+ channels often have a restricted distribution over the elaborate endoplasmic reticulum (ER) in neuronal cells; however, the mechanisms maintaining Ca2+ channel position within the ER membrane are elusive. By means of confocal immunofluorescence microscopy, ryanodine receptor (RyR) Ca2+ channels were localized in honeybee photoreceptors, and the role of actin filaments in RyR positioning was examined. In control cells, RyRs were enriched at distinct microdomains of the ER next to the photoreceptive microvilli. After incubation with cytochalasin B or cytochalasin D, which results in a disruption of F-actin, RyRs were distributed through the entire ER, suggesting that RyRs are positioned within honeybee visual cells by interaction with the F-actin system. PMID- 11406326 TI - Vaginal hyperalgesia in a rat model of endometriosis. AB - This study examined whether a rat model of surgically-induced endometriosis that reduces fertility also evokes vaginal hyperalgesia along with changes in vaginal compliance. In nine rats trained to escape vaginal distention, percent escape responses to different volumes of vaginal distention were measured for 2.5 months before and after endometriosis or sham surgery. Vaginal pressures were also measured simultaneously to provide an estimate of vaginal compliance. Endometriosis (or sham) was induced by autotransplantation of small pieces of uterus (or fat) on mesenteric cascade arteries, abdomen, and ovary. Escape responses were significantly increased only in rats whose autotransplants had formed cysts. Vaginal pressures, however, remained unchanged. This vaginal hyperalgesia may involve a process of viscero-visceral referred hyperalgesia. PMID- 11406327 TI - Compensatory visual field training for patients with hemianopia after stroke. AB - Twenty-one patients with hemianopia received 4 weeks of compensatory visual field training. Detection of and reaction time to visual stimuli were measured with eyes fixating (condition A) and with use of exploratory eye movements (condition B) before and after training. Twenty-three healthy individuals served as control subjects for measurements of parameters during both conditions. Patients with hemianopia to either side showed a marked improvement of detection and reaction time during condition B, but minimum or no change during condition A. Improvements were maintained 8 months after training. Activity of daily living skills also improved in all patients. The size of scotoma on computerized perimetry, in contrast, remained unchanged. Training improved detection of and reaction to visual stimuli without restitution of the visual field defect. PMID- 11406328 TI - Are the estrogen receptors involved in Alzheimer's disease? AB - Retrospective analysis shows that women who took estrogen replacement therapy may have less risk of cognitive decline and of developing Alzheimer's disease (AD). The greater risk associated with female gender and these observations suggest that estrogen may be implicated in the aetiology of AD. Estrogen is one of a family of sex steroids that exerts many of its genomic effects through the activation of the nuclear estrogen receptors, ERalpha and ERbeta. Previously, increased risk for AD has been reported for polymorphisms in the ERalpha gene in a Japanese cohort, however, this association has not been systematically replicated. We have further investigated polymorphisms in the ERalpha and have extended this to investigate an association with a polymorphism within the ERbeta gene in an independent UK Caucasian population. We found no independent association of these polymorphisms with the risk of developing AD in the total sample nor within either gender. However, we did detect a significant interaction between the ERalpha and ERbeta polymorphisms and the risk for AD (OR=0.22 95% CI (0.05-0.88), P=0.02). If this finding can be supported in other independent studies, it may suggest that the risk for AD may be modulated only when both ERalpha and ERbeta have particular variations in their expression and/or biological activities. PMID- 11406329 TI - Bax, Bcl-2, and cyclin expression and apoptosis in rat substantia nigra during development. AB - Naturally occurring cell death via apoptosis has been reported in the substantia nigra of rats during development, culminating during the perinatal period. Cellular pathways leading to apoptotic death of developing nigral dopamine neurons remain unknown, although the apoptotic mediator, caspase 3, has been shown to be activated during this process. Our previous results demonstrated the inability of antioxidants to rescue the nigral dopamine neurons that undergo apoptosis during development. In the present study, we investigated using immunohistochemistry the expression of cyclins D1, D3, and E in the substantia nigra during pre- and postnatal development, since their re-expression in postmitotic neurons has been proposed to contribute to developmental apoptosis. We also investigated by Western blot analysis of nigral tissue isolated during the first postnatal week the expression of the anti- and pro-apoptotic proteins, Bcl-2 and Bax, respectively, since altered Bcl-2 expression during developmental apoptosis has been described. During apoptotic death of nigral dopamine neurons in development, we detected a significant increase in the Bax:Bcl-2 ratio, which is consistent with enhanced apoptosis. There were no changes in the expression of the cyclins during the same apoptotic period. These novel findings suggest that nigral dopamine neurons undergo developmental apoptotic death through a Bax:Bcl-2 sensitive pathway that does not involve cyclin mediation. PMID- 11406330 TI - Presenilin mutations line up along transmembrane alpha-helices. AB - Presenilin 1 mutations are the major cause of autosomal dominant Alzheimer's disease. Here we present evidence that pathogenic mutations in putative transmembrane domains 1, 2, 3, 4 and 6 align along helical faces, thus supporting the view that these are indeed transmembrane domains, and suggesting that disruption of the alignment of these domains is responsible for the pathogenicity of the mutations. PMID- 11406331 TI - Increased hydroxyl radicals in the hippocampus of stroke-prone spontaneously hypertensive rats during transient ischemia and recirculation. AB - It has been suggested that stroke-prone spontaneously hypertensive rats (SHRSP) show vulnerability to neuronal damage following transient ischemia. To observe the effect of hydroxyl radicals on neuronal damage in the hippocampus of SHRSP during ischemia and recirculation, we measured the levels of 2,3-dihydroxybenzoic acid (2,3-DHBA), as a biological marker of hydroxyl radicals in the hippocampus of SHRSP, by high pressure liquid chromatography-electrochemical detection. The production of hydroxyl radicals in the hippocampus during the first 20 min of recirculation was a peak in all intervals. The changes in 2,3-DHBA levels during ischemia and recirculation in SHRSP were significantly higher than in Wistar Kyoto rats. These results suggest that neuronal damage following ischemia and recirculation is, in part, caused by the increase in hydroxyl radicals during ischemia and recirculation. PMID- 11406332 TI - Zoster-associated pain and neural dysfunction. PMID- 11406333 TI - Hypersensitivity to visceral and cutaneous pain in the irritable bowel syndrome. AB - Irritable bowel syndrome (IBS) is one of the most common gastrointestinal illnesses and is characterized by altered visceral perception. Previous studies in IBS have failed to demonstrate altered somatic or cutaneous perception. The aims of the study were to determine whether IBS patients have visceral hypersensitivity and cutaneous heat-induced hyperalgesia restricted to lumbosacral dermatomes, consistent with a localized segmental mechanism. Twelve patients (ten women, two men) with IBS and 17 control subjects (13 women, four men) rated pain intensity and unpleasantness to distension of the rectum (35, 55 mmHg) and thermal stimulation (45, 47 degrees C) of the hand and foot. Patients with IBS demonstrated cutaneous allodynia/hyperalgesia to thermal pain applied to the hand and foot. The cutaneous hyperalgesia was pronounced in the lower extremity yet present in the upper extremity to a lesser extent. Psychological testing revealed the IBS patients report more state anxiety and a greater number of somatic symptoms that significantly correlated with most of the pain measures. However, they did not differ from controls on several personality trait measures. These results suggest that patients with IBS have visceral hyperalgesia and cutaneous hyperalgesia that is distributed over a considerable rostral-caudal distance yet optimally expressed in lumbosacral dermatomes. This distribution is consistent with patterns of spinal hyperexcitability observed in experimentally induced persistent pain conditions. PMID- 11406334 TI - Increased spinal N-methyl-D-aspartate receptor function after 20 h of carrageenan induced inflammation. AB - Spinal N-methyl-D-aspartate (NMDA) receptors are thought to be important in states of central hyperexcitability induced by e.g. inflammation or painful neuropathies. The carrageenan model of inflammatory pain has been and still is widely used as is the NMDA receptor antagonist 2-amino-5-phosphonopentanoic acid (AP5) to investigate NMDA receptor function. Here we present two novel findings using electrophysiological technique: the NMDA receptor function in the spinal cord is increased following 20 h of carrageenan-induced inflammation and further that only the D-isomer of AP5 is active in the spinal cord. Exogenous NMDA (0.5 and 5 nmol) applied onto the dorsal spinal cord produced a significantly greater facilitation and D-AP5 (1.25 micromol) a significantly greater inhibition of the C-fibre evoked response of the wide dynamic range (WDR) neurones studied in carrageenan (20 h after injection) compared to control rats. The present and two recent studies suggest central changes are different and possibly greater in the later (20 h) compared to the earlier (2-6 h) phase of carrageenan-induced inflammation. In conclusion, 20 h of carrageenan-induced inflammation increases the function of spinal NMDA receptor involved in nociceptive transmission and in addition the D-isomer of AP5 should be used when NMDA receptor antagonism is wanted in the spinal cord. PMID- 11406335 TI - Analgesic profile of peroral and topical ketoprofen upon low pH-induced muscle pain. AB - Topical analgesics are widely marketed for treatment of muscle and joint pain. We have recently developed a model of muscle pain and have used this model to evaluate the efficacy of commercially available topical and peroral ketoprofen in order to evaluate the time- and dose-dependence of analgesia. In the present study, we examined the dose- (0, 50, and 100 mg) and time-dependence (hourly to 8 h) of commercially available peroral and topical ketoprofen. In order to achieve infusion times of 8 h (and thus study the time course of analgesic action), we adapted the model of low pH-induced muscle pain in humans to these requirements by applying the infusions continuously for 10 min every hour for 8 h. We found that the 10 min infusion produced reliable and consistent pain levels that were reproducible over the 8 h of the study. The study was performed double-blind, randomized, and with a 1-week interval between each of five different sessions (cross-over). Altogether six volunteers underwent intramuscular infusions of isotonic phosphate-buffered saline solution of pH 5.2; during each 8 h session the infusion was switched on eight times with a duration of 10 min at 50 min intervals (there was no infusion during the 50 min interval). The intramuscular infusion of low pH phosphate buffer induced a localized dull-aching or stinging muscle pain sensation; the flow rate of the pH infusion was individually adjusted to induce pain of a magnitude of 20% on a visual analogue scale (ranging from "no pain" (0%) to "unbearable pain" (100%)). Twenty minutes after starting the infusion the volunteers received a capsule with either a placebo or 50 or 100 mg ketoprofen perorally and, in addition, either placebo gel or 50 or 100 mg of a 2.5% commercial ketoprofen gel was applied topically to the skin. One of the sessions included a placebo gel and an oral placebo. The intensity of the recurrent pain stimulus was significantly reduced by 59% following administration of 100 mg peroral ketoprofen within the first 3 h (P<0.03, Wilcoxon test); this analgesia lasted up to the sixth hour of the experimental protocol. Oral ketoprofen (50 mg) was less effective and reduced the pain intensity by 45% (P<0.05) from only the second to the third hour. In contrast, pain reduction after topical ketoprofen application was not of the same magnitude but appeared to be faster to develop (with a maximum effect within 1 h) on average. The maximum pain suppression with 100 mg topical 2.5% ketoprofen gel was by 51% (significant with P<0.03), while 50 mg topical ketoprofen produced a non significant reduction of 29%. The apparent analgesia was rapid to develop but transient and pain ratings increased back to baseline values within 3 h for the 100 mg dose and within 2 h for the 50 mg dose. This data suggests that topical application of commercial gel-based systems does not provide long-lasting analgesia in the muscle when compared to perorally-dosed ketoprofen. In addition, the data show that even doses of 100 mg peroral ketoprofen do not provide complete relief of muscle pain. PMID- 11406336 TI - Role of calcium channels in the spinal transmission of nociceptive information from the mesentery. AB - Opioids, alpha(2)-adrenoceptor agonists and blockers of voltage-gated calcium channels (VGCCs) have been attributed antinociceptive activity in various experimental set-ups. The present study tested the ability of morphine, clonidine and drugs acting at various VGCCs to inhibit the transmission of noxious stimuli from the mesentery at the level of the spinal cord. In rats under barbiturate anaesthesia traction of 20 g was applied to a bundle of mesenteric blood vessels. This caused immediate transient changes of mean arterial pressure that were taken as indication of nociception. Similar reflexes were elicited by applying 0.6% acetic acid to the same bundle of vessels. The reflexes were dose-dependently reduced by intrathecal administration of morphine or clonidine, but were left unaltered by intrathecal administration of verapamil, Bay-K 8644 or omega conotoxin MVIIA. Neither verapamil nor Bay-K 8644 influenced clonidine-induced analgesia. Conotoxin markedly enhanced the effectiveness of all doses of clonidine against both types of mesenteric stimuli. Verapamil, Bay-K 8644, as well as conotoxin reduced the ability of morphine to inhibit mechanically evoked reflexes, while there was no statistically significant effect in chemonociception. These data suggest that, at the spinal level, both morphine and clonidine are effective drugs to decrease the cardiovascular changes caused by acute mesenteric pain. In the dorsal spinal cord neither L-type nor N-type VGCCs are responsible on their own for the transmission of noxious stimuli from the mesentery. Inhibition of N-type channels markedly augments the action of clonidine, whereas blocking either VGCC seems to inhibit antinociceptive mechanisms induced by morphine. It is suggested that in patients the combined administration of clonidine with omega-conotoxin MVIIA might lead to effective pain control with reduced side effects. PMID- 11406337 TI - Spinal NK1 receptor is upregulated after chronic bladder irritation. AB - It has been suggested that there is a significant upregulation of the NK1 receptor (NK1R) on neurons in the dorsal spinal cord after long-term somatic inflammation. This upregulation appears to play a significant role in central sensitization in chronic pain states. However, it is not clear whether such a change is also observed after chronic visceral (bladder) inflammation. Changes in NK1R immunoreactivity after chronic bladder irritation were investigated in order to evaluate the existence of hypersensitive states in the spinal cord after chronic bladder irritation. Experiments were performed on a total of 12 adult female Sprague-Dawley rats. In six animals, cyclophosphamide (CPA) was administered intraperitoneally for 2 weeks. Another six animals were given intraperitoneal saline injections and served as the control group. After these treatments, immunohistochemical staining for NK1Rs and substance P in rat lumbosacral spinal cord was performed. In CPA-treated animals, NK1R-positive areas and staining intensity within the dorsal spinal cord were significantly increased in the L5 to S2 spinal cord areas, especially in the L6 and S1 segments. In the L6 spinal segment, CPA-treatment enhanced NK1R immunostaining in the medial and the lateral dorsal horn, as well as in the lateral laminae including the sacral parasympathetic nucleus to a lesser extent. In CPA-treated animals, substance P staining intensity increased in the same regions in which NK1R immunoreactivity was increased. This finding probably implies the upregulation of spinal NK1R and the occurrence of central sensitization within the spinal cord after chronic visceral inflammation. PMID- 11406338 TI - Antinociception by tricyclic antidepressants in the rat formalin test: differential effects on different behaviours following systemic and spinal administration. AB - The present study (1) examined analgesic effects of systemically and spinally administered antidepressants (ADs) on phase 2 flinching and biting/licking behaviours in the rat formalin test, a model considered to be of greater relevance to clinical pain than acute threshold tests, and (2) determined whether motor or anti-inflammatory effects contributed to such actions. Systemic administration of amitriptyline (3-20 mg/kg) produced a dose-related enhancement of flinching behaviours, while at the same time suppressing biting/licking behaviours. Imipramine (except for 20 mg/kg), nortriptyline, desipramine and fluoxetine had no significant effect on flinching behaviours, while producing a dose-related suppression of biting/licking behaviours. When administered spinally, either by acute lumbar puncture or via chronically implanted intrathecal cannulas, amitriptyline similarly augmented flinching behaviours. When given by lumbar puncture, amitriptyline suppressed biting/licking behaviours, but when intrathecal cannulas were used, this behaviour was not expressed in the formalin group. Other ADs also suppressed biting/licking behaviours without affecting flinching when given by lumbar puncture. Effects on paw volume were determined at the end of behavioural testing. Systemic administration of all ADs produced a dose-related reduction in paw volume. Spinal administration of nortriptyline by lumbar puncture also reduced paw volume, but for other agents, the reduction was not significant. Motor effects were noted qualitatively throughout these experiments, and considered in relation to nociceptive behaviours. These results indicate (a) a marked dissociation between the effects of systemic ADs on flinching and biting/licking behaviours in the formalin test, (b) spinal efficacy of ADs that essentially reproduces effects seen with systemic administration when given by lumbar puncture, (c) a lack of causality between anti-inflammatory effects of ADs and their analgesic properties in the formalin test, and (d) a contribution of motor effects to analgesic actions at higher doses affecting biting/licking but not flinching behaviours. PMID- 11406339 TI - L-type and T-type calcium channel blockade potentiate the analgesic effects of morphine and selective mu opioid agonist, but not to selective delta and kappa agonist at the level of the spinal cord in mice. AB - We examined the effects of amlodipine, a selective L-type voltage dependent Ca(2+) channel (VDCC) blocker, and mibefradil, a selective T-type VDCC blocker on the antinociceptive effects of morphine, and mu, delta and kappa opioid receptor selective agonist-induced antinociception at the spinal level. Intrathecally administered amlodipine and mibefradil potentiated morphine and [D-Ala(2), N mePhe(4), Gly-ol(5)] enkephalin (DAMGO)-induced antinociception by shifting their dose response curves to the left. However, intrathecally administered amlodipine and mibefradil did not affect [D-Pen(2), D-Pen(5)]enkephalin (DPDPE) and [trans (+/-)-3,4-dichloro-N-methyl-N-[2-(1-pyrolidinyl)cyclohexyl] benzene acetamide (U 50, 488H)-induced antinociception. These data indicate that L-type and T-type VDCC blockers synergistically potentiate the analgesic effects of mu opioid receptor agonists, but not delta and kappa opioid receptor agonists, at the spinal level. Additionally, these data suggest that there is an important functional interaction between L-type and/ or T-type VDCC and mu opioid receptors in the process of analgesia. PMID- 11406340 TI - Vincristine-induced allodynia in the rat. AB - The aims of this study were two-fold: first, to simplify the method for creating a recently described neuropathic pain model in the rat, and second, to evaluate the effects of a number of drugs with analgesic or antihyperalgesic properties, in this model. Continuous intravenous vincristine infusion (1-100 microg kg(-1) day (-1)) for 14 days resulted in a dose dependent tactile allodynia (as measured by von Frey filaments) by 7 days at doses between 30 - 100 microg kg(-1) day ( 1), with a hindlimb motor deficit observed at doses greater than 50 microg kg(-1) day (-1). No thermal hyperalgesia was observed. Systemic morphine, lidocaine, mexiletine and pregabalin (given intraperitoneally) produced significant reduction of the allodynia, while tetrodotoxin was without effect. Continuous intravenous infusion of vincristine in rats thus provides a reliable model of chemotherapy induced neuropathy which may be used in defining the mechanism and pharmacology of this clinically relevant condition. PMID- 11406341 TI - Response expectancies in placebo analgesia and their clinical relevance. AB - Response expectancies have been proposed as the major determinant of placebo effects. Here we report that different expectations produce different analgesic effects which in turn can be harnessed in clinical practice. Thoracotomized patients were treated with buprenorphine on request for 3 consecutive days, together with a basal intravenous infusion of saline solution. However, the symbolic meaning of this basal infusion was changed in three different groups of patients. The first group was told nothing about any analgesic effect (natural history). The second group was told that the basal infusion was either a powerful painkiller or a placebo (classic double-blind administration). The third group was told that the basal infusion was a potent painkiller (deceptive administration). Therefore, whereas the analgesic treatment was exactly the same in the three groups, the verbal instructions about the basal infusion differed. The placebo effect of the saline basal infusion was measured by recording the doses of buprenorphine requested over the three-days treatment. We found that the double-blind group showed a reduction of buprenorphine requests compared to the natural history group. However, this reduction was even larger in the deceptive administration group. Overall, after 3 days of placebo infusion, the first group received 11.55 mg of buprenorphine, the second group 9.15 mg, and the third group 7.65 mg. Despite these dose differences, analgesia was the same in the three groups. These results indicate that different verbal instructions about certain and uncertain expectations of analgesia produce different placebo analgesic effects, which in turn trigger a dramatic change of behaviour leading to a significant reduction of opioid intake. PMID- 11406342 TI - Gabapentin actions on N-methyl-D-aspartate receptor channels are protein kinase C dependent. AB - Gabapentin (Neurontin) (GBP) is a widely prescribed analgesic used in treating pain patients with peripheral nerve injuries, diabetic neuropathy and cancer. To understand the mechanism of its action, we used the whole-cell patch recording technique to study the effects of GBP on N-methyl-D-aspartate (NMDA)-evoked currents in single dorsal horn neurons isolated from normal rats and from rats with inflammation induced by the injection of complete Freund adjuvant (CFA) to the hindpaw. We found that GBP enhanced NMDA currents in normal neurons only when protein kinase C (PKC) was added to these cells. The enhancement resulted from an increase in the affinity of glycine for NMDA receptors by GBP. In contrast, in neurons isolated from CFA-treated rats, GBP enhanced NMDA responses without any PKC treatment. Since endogenous PKC in inflamed tissue is elevated, these results suggest that GBP exerts its effects only on those cells affected by inflammatory injuries. Thus, the effects of GBP on NMDA receptors are plastic; they depend on the phosphorylation states of cells or receptors. These observations point to a new strategy for drug design. A chemical whose action depends on the state of cells would maximize its effectiveness while keeping its side-effects to a minimum. PMID- 11406343 TI - The unfolded protein response and Alzheimer's disease. AB - Disruption of calcium homeostasis, inhibition of protein glycosylation, and reduction of disulfide bonds provoke accumulation of unfolded protein in the endoplasmic reticulum (ER), and are therefore a type of 'ER stress'. Normal cells respond to ER stress by increasing transcription of genes encoding ER-resident chaperones such as GRP78/BiP, GRP94 and protein disulfide isomerase to facilitate protein folding. This induction system is termed the unfolded protein response. Familial Alzheimer's disease-linked presenilin-1 (PS1) mutation downregulates the unfolded protein response and leads to vulnerability to ER stress. The mechanisms by which mutant PS1 affects the ER stress response are attributed to the inhibited activation of ER stress transducers such as IRE1, PERK and ATF6. PMID- 11406344 TI - The facile detection of 1505G-->A in Gaucher patients with different phenotypes. AB - In Gaucher disease patients, over 100 disease-causing mutations have been identified. For identification of the 1504C-->T (R463C) mutation it is common to use PCR-restriction fragmentation analysis using the restriction enzyme MspI. In the present study we investigated the reliability of this approach because accurate determination of genotypes is important in genotype-phenotype correlations. A simple modification, i.e. using the restriction enzyme HphI instead of MspI, revealed that type I and II Gaucher disease patients who had previously been identified as carrying the 1504C-->T mutation in fact carried the 1505G-->A (IVS10(-1)G-->A) mutation. Sequencing of the appropriate fragment confirmed this. The PCR method easily differentiates between these two mutations in Gaucher disease patients, thus circumventing the need for sequencing procedures. The phenotypes of the patients found to be carrying the 1505G-->A mutation are also described. PMID- 11406345 TI - Intracellular pH regulation in colonocytes of rat proximal colon. AB - The regulation of intracellular pH (pH(i)) in colonocytes of the rat proximal colon has been investigated using the pH-sensitive dye BCECF and compared with the regulation of pH(i) in the colonocytes of the distal colon. The proximal colonocytes in a HEPES-buffered solution had pH(i)=7.24+/-0.04 and removal of extracellular Na(+) lowered pH(i) by 0.24 pH units. Acid-loaded colonocytes by an NH(3)/NH(4)(+) prepulse exhibited a spontaneous recovery that was partially Na(+) dependent and could be inhibited by ethylisopropylamiloride (EIPA). The Na(+) dependent recovery rate was enhanced by increasing the extracellular Na(+) concentration and was further stimulated by aldosterone. In an Na(+)- and K(+) free HEPES-buffered solution, the recovery rate from the acid load was significantly stimulated by addition of K(+) and this K(+)-dependent recovery was partially blocked by ouabain. The intrinsic buffer capacity of proximal colonocytes at physiological pH(i) exhibited a nearly 2-fold higher value than in distal colonocytes. Butyrate induced immediate colonocyte acidification that was smaller in proximal than in distal colonocytes. This acidification was followed by a recovery phase that was both EIPA-sensitive and -insensitive and was similar in both groups of colonocytes. In a HCO(3)(-)/CO(2)-containing solution, pH(i) of the proximal colonocytes was 7.20+/-0.04. Removal of external Cl(-) caused alkalinization that was inhibited by DIDS. The recovery from an alkaline load induced by removal of HCO(3)(-)/CO(2) from the medium was Cl(-)-dependent, Na(+) independent and blocked by DIDS. Recovery from an acid load in EIPA-containing Na(+)-free HCO(3)(-)/CO(2)-containing solution was accelerated by addition of Na(+). Removal of Cl(-) inhibited the effect of Na(+). In summary, the freshly isolated proximal colonocytes of rats express Na(+)/H(+) exchanger, H(+)/K(+) exchanger ((H(+)-K(+))-ATPase) and Na(+)-dependent Cl(-)/HCO(3)(-) exchanger that contribute to acid extrusion and Na(+)-independent Cl(-)/HCO(3)(-) exchanger contributing to alkali extrusion. All of these are likely involved in the regulation of pH(i) in vivo. Proximal colonocytes are able to maintain a more stable pH(i) than distal cells, which seems to be facilitated by their higher intrinsic buffer capacity. PMID- 11406346 TI - Model for the structure of the HIV gp41 ectodomain: insight into the intermolecular interactions of the gp41 loop. AB - In human immunodeficiency virus (HIV) the viral envelope proteins gp41 and gp120 form a non-covalent complex, which is a potential target for AIDS therapies. In addition gp41 plays a possible role in HIV infection of B cells via the complement system. In an effort to better understand the molecular interactions of gp41, the structure of the HIV gp41 ectodomain has been modeled using the NMR restraints of the simian immunodeficiency virus (SIV) gp41 ectodomain (M. Caffrey, M. Cai, J. Kaufman, S.J. Stahl, P.T. Wingfield, A.M. Gronenborn, G.M. Clore, Solution structure of the 44 kDa ectodomain of SIV gp41, EMBO J. 17 (1998) 4572--4584). The resulting model presents the first structural information for the HIV gp41 loop, which has been implicated to play a direct role in binding to gp120 and C1q of the complement system. PMID- 11406347 TI - Plasma factors controlling atrial natriuretic peptide (ANP) aggregation: role of lipoproteins. AB - We have previously shown that human plasma atrial alpha-natriuretic peptide (alpha-hANP) sequestering is a protective phenomenon against amyloid aggregation. In the present work, the possible role of lipoproteins as alpha-hANP binding factors has been investigated in vitro using an experimental model, developed in our laboratory, that allows to work at physiological concentrations. This approach consists of gel filtration on Sephacryl S-300 HR of big alpha [(125)I]hANP generated in phosphate buffered saline or in human normal plasma supplemented or not with lipoproteins. The results of these experiments indicate that high density lipoproteins (HDL) are responsible for the ANP binding phenomenon observed in vitro, while low density lipoproteins and very low density lipoproteins do not directly interact with ANP. Moreover, the HDL remodeling process occurring in vitro has been analyzed during plasma incubation by monitoring the redistribution of lipids and apolipoproteins among the HDL subclasses. The changes in HDL size and composition observed in incubated plasma were compared with the redistribution of endogenous and labeled big ANP. The obtained results revealed that both tend to follow the molecular rearrangement in plasma of apolipoprotein A-I containing particles and suggested that, among HDL species, the small particles are mainly involved in the ANP binding phenomenon. This hypothesis was further demonstrated by ligand blotting experiments that confirmed the existence of differences in the ability of HDL particles to bind alpha-[(125)I]hANP. PMID- 11406348 TI - Development of potential iron chelators for the treatment of Friedreich's ataxia: ligands that mobilize mitochondrial iron. AB - Friedreich's ataxia (FA) is a crippling neurodegenerative disease that is due to iron (Fe) overload within the mitochondrion. One therapeutic intervention may be the development of a chelator that could remove mitochondrial Fe. We have implemented the only well characterized model of mammalian mitochondrial Fe overload to examine the Fe chelation efficacy of novel chelators of the 2 pyridylcarboxaldehyde isonicotinoyl hydrazone (PCIH) class. In this model we utilize reticulocytes treated with the haem synthesis inhibitor succinylacetone which results in mitochondrial Fe-loading. Our experiments demonstrate that in contrast to desferrioxamine, several of the PCIH analogues show very high activity at mobilizing (59)Fe from (59)Fe-loaded reticulocytes. Further studies on these ligands in animals are clearly warranted considering their potential to treat FA. PMID- 11406349 TI - A missense mutation in the Na(+)/glucose cotransporter gene SGLT1 in a patient with congenital glucose-galactose malabsorption: normal trafficking but inactivation of the mutant protein. AB - The Na(+)/glucose cotransporter gene SGLT1 was analyzed in a Japanese patient with congenital glucose-galactose malabsorption. Genomic DNA was used as a template for amplification by the polymerase chain reaction of each of the 15 exons of SGLT1. The amplification products were cloned and sequenced. About half of the exon 5 clones of the patient contained a C-->T transition, resulting in an Arg(135)-->Trp mutation, whereas the remaining clones contained the normal exon 5 sequence. In addition, whereas some exon 12 clones exhibited the normal sequence, others showed a CAgtaggtatcatc-->CAgacc mutation at the splice donor site of intron 12 that may result either in the skipping of exon 12 or in read-through of intron 12. Neither the Arg(135)-->Trp mutant nor either of the possible intron 12 mutant proteins exhibited Na(+)-dependent glucose transport activity when expressed in Xenopus oocytes. Immunocytochemical analysis indicated, however, that the Arg(135)-->Trp mutant was localized to the oocyte plasma membrane. DNA sequence analysis revealed that the missense mutation in exon 5 and the splice site mutation in intron 12 were inherited from the proband's father and mother, respectively. These results indicate that the patient is a compound heterozygote for this disease, and that the Arg(135)-->Trp mutant of SGLT1 undergoes normal trafficking to the plasma membrane but is non-functional. PMID- 11406350 TI - Suppression of a sialyltransferase by antisense DNA reduces invasiveness of human colon cancer cells in vitro. AB - Transfer of terminal alpha 2,6-linked sialic acids to N-glycans is catalyzed by beta-galactoside alpha 2,6-sialyltransferase (ST6Gal I). Expression of ST6Gal I and its products is reportedly increased in colon cancers. To investigate directly the functional effects of ST6Gal I expression, human colon cancer (HT29) cells were transfected with specific antisense DNA. ST6Gal I mRNA and protein were virtually undetectable in six strains of transfected HT29 cells. ST6Gal activity was reduced to 14% of control (P<0.005) in transfected cells. Expression of terminal alpha 2,6- and alpha 2,3-linked sialic acids, and unmasked N acetyllactosamine oligosaccharides, respectively, was assessed using flow cytometry and fluoresceinated Sambucus nigra, Maackia amurensis and Erythrina cristagalli lectins. Results indicated a major reduction in expression of alpha 2,6-linked sialic acids and counterbalancing increase in unmasked N acetyllactosamines in antisense DNA-transfected cells, without altered expression of alpha 2,3-linked sialic acids or ganglioside profiles. The ability of transfected cells to form colonies in soft agar and to invade extracellular matrix material (Matrigel), respectively, in vitro was reduced by approx. 98% (P<0.0001) and more than 3-fold (P<0.005) compared to parental HT29 cells. These results indicate that N-glycans bearing terminal alpha 2,6-linked sialic acids may enhance the invasive potential of colon cancer cells. PMID- 11406351 TI - The polycystin-1 C-type lectin domain binds carbohydrate in a calcium-dependent manner, and interacts with extracellular matrix proteins in vitro. AB - Mutations in the PKD1 gene are responsible for 85% of cases of autosomal dominant polycystic kidney disease (ADPKD). This gene encodes a large membrane associated glycoprotein, polycystin-1, which is predicted to contain a number of extracellular protein motifs, including a C-type lectin domain between amino acids 403--532. We have cloned and expressed the PKD1 C-type lectin domain, and have demonstrated that it binds carbohydrate matrices in vitro, and that Ca(2+) is required for this interaction. This domain also binds to collagens type I, II and IV in vitro. This binding is greatly enhanced in the presence of Ca(2+) and can be inhibited by soluble carbohydrates such as 2-deoxyglucose and dextran. These results suggest that polycystin-1 may be involved in protein-carbohydrate interactions in vivo. The data presented indicate that there may a direct interaction between the PKD1 gene product and an ubiquitous extracellular matrix (ECM) protein. PMID- 11406352 TI - Experimental evidence suggesting that nitric oxide diffuses from tissue into blood but not from blood into tissue. AB - The aim of this study was to evaluate in vivo whether nitric oxide (NO) is able to diffuse from blood into tissues and vice versa from tissues into blood. We used an in vivo model of intestinal ischemia (superior mesenteric artery occlusion) selectively increasing NO levels in intestinal tissue and an infusion of L-arginine selectively increasing NO levels in blood. In this model we followed formation of nitrosyl complexes of hemoglobin (Hb-NO) in blood and nitrosyl-diethyldithiocarbamate-iron complexes (DETC--Fe--NO) in ischemic intestine and normoxic tissues by means of electron paramagnetic resonance spectroscopy. NO trapping by DETC--Fe in the tissues resulted in a reduction of Hb--NO levels in blood accompanied by the formation of water-insoluble DETC--Fe NO complexes in ischemic intestine and normoxic tissues both during ischemia and during reperfusion. Administration of L-arginine increased NO levels in blood but neither in ischemic intestine nor in normoxic tissue. Our data suggest that NO released in blood from endothelial cells does not diffuse into tissue. In contrast, NO formed in tissue diffuses into blood. The latter indicates that NO formed in tissues may exert its biological activities systematically. PMID- 11406353 TI - Antifibrogenic effect in vivo of low doses of insulin-like growth factor-I in cirrhotic rats. AB - Insulin-like growth factor-I (IGF-I) is produced mainly in the liver and it induces beneficial effects on the nutritional status, the liver function and oxidative hepatic damage in cirrhotic rats. The aim of this work was to analyze the effect of IGF-I on mechanisms of fibrogenesis in cirrhotic rats. Liver cirrhosis was induced by CCl(4) inhalation and phenobarbital in Wistar rats. Ten days after stopping CCl(4) administration (day 0), rats received either IGF-I (2 microg/100 g bw/day) (CI+IGF) or saline (CI) subcutaneously during 14 days. Animals were sacrificed on day 15. As control groups were used: healthy rats (CO) and healthy rats treated with IGF-I (CO+IGF). Liver histopathology, hydroxyproline content, prolyl hydroxylase activity, collagen I and III mRNA expression and the evolution of transformed Ito cells into myofibroblasts were assessed. Among the two control groups (CO+IGF), no differences were found in hydroxyproline content and these levels were lower than those found in the two cirrhotic groups. Compared with untreated cirrhotic rats, the CI+IGF-I animals showed a significant reduction in hydroxyproline content, prolyl hydroxylase activity and collagen alpha 1(I) and alpha1(III) mRNA expression. A higher number of transformed Ito cells (alpha-actin +) was observed in untreated cirrhotic animals as compared to CO and CI+IGF groups. In summary, treatment with IGF-I reduced all of the studied parameters of fibrogenesis. In conclusion, low doses of IGF-I induce in vivo an antifibrogenic effect in cirrhotic rats. PMID- 11406355 TI - Web alert. Lymphocyte activation and effector functions. PMID- 11406357 TI - New views of BCR structure and organization. AB - Recent work has provided new insights into the stoichiometry of BCR subunits, as well as the organization of the BCR before and after engagement by antigen. On resting cells, the BCR may be pre-assembled into oligomeric receptor complexes that generate a basal level of signaling. After antigen binding, the BCR may be organized into larger receptor arrays that reside in lipid rafts - sites where signaling enzymes are concentrated. The critical role of BCR assembly and organization in B cell function is underscored by the recent findings that this process is altered in many B cell tumors. PMID- 11406358 TI - B lymphocyte activation by contact-mediated interactions with T lymphocytes. AB - T cell dependent B lymphocyte activation requires interactions between numerous receptor-ligand pairs on the two cell types. Recently, advances have been made both in understanding how these various signals regulate B cell effector functions and in identifying many new receptor-ligand pairs that contribute to the regulation of B cell function by T lymphocytes. PMID- 11406359 TI - Reprogramming T cells: the role of extracellular matrix in coordination of T cell activation and migration. AB - The stable immunological synapse between a T cell and antigen-presenting cell coordinates migration and activation. Three-dimensional collagen gels transform this interaction into a series of transient hit-and-run encounters. Here we integrate these alternative modes of interaction in a model for primary T cell activation and effector function in vivo. PMID- 11406360 TI - The instructive role of dendritic cells on T cell responses: lineages, plasticity and kinetics. AB - Dendritic cells are responsible for directing different types of T cell responses, from thymic negative selection to the generation of effector and memory cells and the induction of peripheral tolerance. Recent studies indicate that the dendritic cell lineage, the extent of recruitment into inflamed tissues and migration to lymph nodes, the nature of maturation stimuli and the kinetics of activation have a quantitative and qualitative impact on T cell stimulation, thus exerting an instructive control on T cell responses. PMID- 11406361 TI - Proximal protein tyrosine kinases in immunoreceptor signaling. AB - Immunoreceptor engagement results in the sequential activation of several classes of protein tyrosine kinases, including the Src and Syk/Zap-70 families. Recent progress has been made in our understanding of the regulation and function of these molecules. First, it was revealed that membrane compartmentation of protein tyrosine kinases may be essential for their proper biological function. Second, Src family kinases were found to act not only as positive regulators, but also as inhibitors of cell activation. Third, it was appreciated that Csk, a potent inhibitor of Src kinases, is regulated by an assortment of protein-protein interactions. Fourth, differences in the regulation of Syk and Zap-70 were observed, suggesting significant distinctions in the purpose of these two kinases in immunoreceptor signaling. And fifth, it was suggested that proximal kinases implicated in immunoreceptor-mediated signal transduction may be regulated by protein degradation via binding to c-Cbl, a ubiquitin ligase. PMID- 11406362 TI - Adapters in lymphocyte signalling. AB - Adapter proteins are well recognised as important molecular switches connecting immunoreceptors with intracellular signalling pathways. However, recent data suggest that homeostasis within the lymphatic system also depends on the coordinated activities of negative regulatory adapter proteins. These prevent activation of lymphocytes in the absence of externally applied signals and regulate termination/limitation of ongoing immune responses via different mechanisms. PMID- 11406363 TI - Tec kinases: modulators of lymphocyte signaling and development. AB - The Tec kinases are implicated as important components of the antigen receptor signaling required for proper lymphocyte activation and development. Recent data suggest that these kinases contribute to multiprotein complexes containing LAT and SLP-76 in T cells, and BLNK/SLP-65 in B cells, which are required for activation of PLC-gamma and downstream pathways. PMID- 11406364 TI - Face off--the interplay between activating and inhibitory immune receptors. AB - The function of leukocytes is regulated by the integration of positive and negative signals received through cell surface receptors. Related receptors with similar extracellular domains and often binding the same ligands can transmit either inhibitory or activating signals. Studies are beginning to reveal how these 'paired receptors' control immune functions. PMID- 11406365 TI - Phosphoinositide 3-kinases in T lymphocyte activation. AB - Biochemical experiments have established that the metabolism of inositol phospholipids by phosphoinositide 3-kinases (PI3Ks) and lipid-phosphatases is triggered by many receptors that control T lymphocyte function, including antigen receptors, costimulatory molecules, cytokines and chemokines. Novel effectors of PI3K have been identified in the immune system and shown to be important in the control of lymphocyte activation. Moreover, key lipid-phosphatases have been identified that act to terminate or modulate PI3K signalling in cells of the immune system. PMID- 11406366 TI - MAP-kinase signaling pathways in T cells. AB - The family of MAP-kinases include ERKs, p38 MAP-kinases and JNKs. Recently, the role of MAP-kinases in T lymphocytes has attracted particular interest. Genetically modified mouse models have brought insight into the specific function of each MAP-kinase pathway in T lymphocyte biology. Studies clearly show that these pathways are not redundant and that the role of each pathway depends on the T cell type and differentiation stage. PMID- 11406367 TI - Monitoring the duration of antigen-receptor occupancy by calcineurin/glycogen synthase-kinase-3 control of NF-AT nuclear shuttling. AB - Recent structural studies have supported a kinetic model of TCR activation, raising the question of how the duration of receptor occupancy is translated into activation of immune response genes. We summarize evidence that the cytoplasmic to-nuclear shuttling of NF-ATc family members monitors the duration of receptor occupancy. PMID- 11406368 TI - Helper T cell differentiation, inside and out. AB - The past year has seen remarkable progress in the field of helper T cell differentiation, including discovery of a novel transcription factor as well as a novel cytokine receptor of the Th1 lineage. The year has also brought new perspectives on the genetic and epigenetic control of gene expression. It is likely that mechanisms of immunity will continue to provide insight into the general problem of cellular decision-making. PMID- 11406369 TI - Activation-induced cell death. AB - The death of T lymphocytes following their activation involves several signal pathways that converge on a series of proteases, known as caspases, that degrade cellular proteins and activate a DNAse. Caspases are activated through ligation of cell surface death receptors as well as via direct activation of downstream caspases, often through metabolic stress such as cytokine withdrawal or generation of oxygen radicals, that culminates in mitochondrial dysfunction and release of the pro-apoptotic molecules, cytochrome c and Smac/DIABLO. The Bcl-2 family members serve to regulate the mitochondrial membrane integrity. Recent studies are now revealing the significant contribution to the activation-induced cell death of T cells by downstream caspases such as caspase-3 and Bcl-2-homology domain 3 (BH3)-only members of the Bcl-2 family. PMID- 11406370 TI - Signaling by type I and II cytokine receptors: ten years after. AB - Discovered during the past ten years, Janus kinases and signal transducers and activators of transcription have emerged as critical elements in cytokine signaling and immunoregulation. Recently, knockout mice for all the members of these families have been generated, with remarkably specific outcomes. Equally exciting is the discovery of a new class of inhibitors, the suppressor of cytokine signaling family. The phenotypes of mice deficient in these molecules are also striking, underscoring the importance of negative regulation in cytokine signaling. PMID- 11406372 TI - Nucleic acids. Sequences and topology Web alert. AB - A selection of World Wide Web sites relevant to reviews published in this issue of Current Opinion in Structural Biology. PMID- 11406374 TI - NMR methodology for the study of nucleic acids. AB - During the past few years, NMR methodology for the study of nucleic acids has benefited from new developments that greatly improved state-of-the-art technology for the precise determination of three-dimensional structures. Substantial progress has been made in designing experimental protocols for the measurement of residual dipolar couplings, in sensitivity optimization of triple-resonance experiments and in detection of hydrogen bonds and in developing computational methods for structure refinement using NMR restraints. PMID- 11406375 TI - Computational methods for RNA structure determination. AB - During the past year, major improvements have been made in methods used to solve RNA structures from crystals, find RNA patterns in sequence data and determine RNA secondary structure. Computational methods for assisting an interactive computer graphics human modeler, searching the conformational space of RNA tertiary structure, studying the dynamics of complexes involving RNA and simulating RNA catalytic activities have also been advanced. PMID- 11406376 TI - Single-molecule fluorescence methods for the study of nucleic acids. AB - Single-molecule fluorescence methods and biomechanical tools provide exciting new opportunities to probe biochemical processes in unprecedented detail. The detection and spectroscopy of single fluorophores have recently been used to observe conformational changes and biochemical events involving nucleic acids. A number of fluorescence observables, including localization, quenching, polarization response and fluorescence resonance energy transfer, have been utilized. An exciting new opportunity of combining fluorescence methods and biomechanical tools to study the structural changes and functions of enzymes that participate in nucleic acid metabolism has also arisen. PMID- 11406377 TI - DNA-cation interactions: The major and minor grooves are flexible ionophores. AB - Several crystallographic, solution-state and theoretical studies carried out this past year provide new support for the sequence-specific nature of monovalent and divalent cation coordination within the DNA major and minor grooves. Correlations observed between groove width and cation coordination indicate that the grooves are flexible and respond to cation binding. PMID- 11406378 TI - The crystal structures of DNA Holliday junctions. AB - Nearly 40 years ago, Holliday proposed a four-stranded complex or junction as the central intermediate in the general mechanism of genetic recombination. During the past two years, six single-crystal structures of such DNA junctions have been determined by three different research groups. These structures all essentially adopt the antiparallel stacked-X conformation, but can be classified into three distinct categories: RNA-DNA junctions; ACC trinucleotide junctions; and drug induced junctions. Together, these structures provide insight into how local and distant interactions help to define the detailed and general physical features of Holliday junctions at the atomic level. PMID- 11406379 TI - Beyond kinetic traps in RNA folding. AB - Large RNAs often have rugged folding energy landscapes that result in severe misfolding and slow folding kinetics. Several interdependent parameters that contribute to misfolding are now well understood and examples of large RNAs and ribonucleoproteins that avoid kinetic traps have been reported. These advances have facilitated the exploration of fundamental RNA folding processes that were previously inaccessible. PMID- 11406380 TI - Structure and function of the small ribozymes. AB - Recently, major advances have been made toward increasing our understanding of small ribozyme structure and function. The first general acid-base catalytic mechanism for a ribozyme has been defined. Shifted nucleotide pK(a) values have been found to be surprisingly frequent structural elements. Finally, the dynamic nature of RNA catalysis has been highlighted through new structural and biochemical information. PMID- 11406381 TI - Artificial ribozymes and deoxyribozymes. AB - RNA and DNA molecules with catalytic properties have been isolated by in vitro selection from combinatorial nucleic acid libraries. A broad range of chemical reactions is catalyzed and nucleic acids can accelerate bond formation between small organic substrates. The catalytic performance of nucleic acids can be enhanced by the incorporation of additional functional groups. PMID- 11406383 TI - Advances in sequence analysis. AB - In its early days, the entire field of computational biology revolved almost entirely around biological sequence analysis. Over the past few years, however, a number of new non-sequence-based areas of investigation have become mainstream, from the analysis of gene expression data from microarrays, to whole-genome association discovery, and to the reverse engineering of gene regulatory pathways. Nonetheless, with the completion of private and public efforts to map the human genome, as well as those of other organisms, sequence data continue to be a veritable mother lode of valuable biological information that can be mined in a variety of contexts. Furthermore, the integration of sequence data with a variety of alternative information is providing valuable and fundamentally new insight into biological processes, as well as an array of new computational methodologies for the analysis of biological data. PMID- 11406384 TI - Clustering and analysis of protein families. AB - Various sequence-motif and sequence-cluster databases have been integrated into a new resource known as InterPro. Because the contributing databases have different clustering principles and scoring sensitivities, the combined assignments complement each other for grouping protein families and delineating domains. InterPro and new developments in the analysis of both the phylogenetic profiles of protein families and domain fusion events improve the prediction of specific functions for numerous proteins. PMID- 11406385 TI - Whole-genome expression analysis: challenges beyond clustering. AB - Measuring the expression of most or all of the genes in a biological system raises major analytic challenges. A wealth of recent reports uses microarray expression data to examine diverse biological phenomena - from basic processes in model organisms to complex aspects of human disease. After an initial flurry of methods for clustering the data on the basis of similarity, the field has recognized some longer-term challenges. Firstly, there are efforts to understand the sources of noise and variation in microarray experiments in order to increase the biological signal. Secondly, there are efforts to combine expression data with other sources of information to improve the range and quality of conclusions that can be drawn. Finally, techniques are now emerging to reconstruct networks of genetic interactions in order to create integrated and systematic models of biological systems. PMID- 11406386 TI - Protein structure similarities. AB - Comparison of protein structures can reveal distant evolutionary relationships that would not be detected by sequence information alone. This helps to infer functional properties. In recent years, many methods for pairwise protein structure alignment have been proposed and are now available on the World Wide Web. Although these methods have made it possible to compare all available protein structures, they also highlight the remaining difficulties in defining a reliable score for protein structure similarities. PMID- 11406387 TI - Determination of protein function, evolution and interactions by structural genomics. AB - The genome sequencing projects and knowledge of the entire protein repertoires of many organisms have prompted new procedures and techniques for the large-scale determination of protein structure, function and interactions. Recently, new work has been carried out on the determination of the function and evolutionary relationships of proteins by experimental structural genomics, and the discovery of protein-protein interactions by computational structural genomics. PMID- 11406388 TI - Protein functional epitopes: hot spots, dynamics and combinatorial libraries. AB - Recent studies increasingly point to the importance of structural flexibility and plasticity in proteins, highlighting the evolutionary advantage. There are an increasing number of cases in which given, presumably specific, binding sites have been shown to bind a range of ligands with different compositions and shapes. These studies have also revealed that evolution tends to find convergent solutions for stable intermolecular associations, largely via conservation of polar residues as hot spots of binding energy. On the other hand, the ability to bind multiple ligands at a given site is largely derived from hinge-based motions. The consideration of these two factors in functional epitopes allows more realism and robustness in the description of protein binding surfaces and, as such, in applications to mutants, modeled structures and design. Efficient multiple structure comparison and hinge-bending structure comparison tools enable the construction of combinatorial binding epitope libraries. PMID- 11406389 TI - Helical membrane proteins: diversity of functions in the context of simple architecture. AB - During the past year, research on helical membrane proteins has brought insights into the use of deviations from canonical alpha-helical conformation to support function and the further investigation of the sequestration of protein regions from the lipid bilayer to enhance these structural alternatives. Also, the structural roles of polar sidechains, the identification of motifs in helix interactions and the significance of certain topologies on a genome-wide scale have been further explored. PMID- 11406390 TI - Whole-genome analysis: annotations and updates. AB - The most important advances in the field of genome annotation over the past two years involve the use of cDNA sequences, protein structures and gene expression data to predict genes. These types of information not only improve gene identification, but they also give insights into variation in gene structure and function. PMID- 11406391 TI - From RNA helicases to RNPases. AB - In eukaryotic cells, all aspects of cellular RNA metabolism require putative RNA helicases of the DEAD and DExH protein families (collectively known as DExD/H families). Based on data from biochemical studies of a few of these RNA helicases, they are generally considered to be involved in the unwinding of duplex RNA molecules. However, recent reports provide evidence indicating that these proteins might also be involved in the active disruption of RNA-protein interactions. PMID- 11406392 TI - The heart of photosynthesis in glorious 3D. AB - The input of solar energy into photosynthesis, and thence into the biosphere, occurs via chlorophyll-containing proteins known as reaction centres. There are two kinds of reaction centre in oxygenic photosynthesis: photosystem I (PSI) and photosystem II (PSII). The PSII reaction centre, alias the oxygen-evolving enzyme, the water-oxidizing complex or the water-plastoquinone photo oxidoreductase, has now been crystallized and its structure solved to a resolution of 3.8 A. PMID- 11406393 TI - The Brix domain protein family -- a key to the ribosomal biogenesis pathway? AB - Six (one archaean and five eukaryotic) protein families have similar domain architecture that includes a central globular Brix domain, and optional N- and obligatory C-terminal segments, both with charged low-complexity regions. Biological data for some proteins in this superfamily suggest a role in ribosome biogenesis and rRNA binding. PMID- 11406394 TI - A ubiquitin-interacting motif conserved in components of the proteasomal and lysosomal protein degradation systems. AB - Ubiquitination generally serves as a signal for targeting cytoplasmic and nuclear proteins to the proteasome for subsequent degradation. Recently, evidence has accumulated indicating that ubiquitination also plays an important role in targeting integral membrane proteins for degradation by the lytic vacuole or the lysosome. This article describes a conserved protein motif, based on a sequence of the proteasomal component Rpn10/S5a, that is known to recognize ubiquitin. The presence of this motif in Eps15, Epsin and HRS, proteins involved in ligand activated receptor endocytosis and degradation, suggest a more general role in ubiquitin recognition. PMID- 11406408 TI - Dynamic Nef and Nef dynamics: how structure could explain the complex activities of this small HIV protein. AB - The Nef protein of the human immunodeficiency virus is as important for disease progression as it is perplexing in its plethora of target molecules and functions. In this article, it is proposed that the complex biology of Nef is regulated through conformational changes of the protein that are triggered by cellular location and specific interactions as Nef traffics through the infected cell. PMID- 11406409 TI - LG/LNS domains: multiple functions -- one business end? AB - The three-dimensional structures of LG/LNS domains from neurexin, the laminin alpha 2 chain and sex hormone-binding globulin reveal a close structural relationship to the carbohydrate-binding pentraxins and other lectins. However, these LG/LNS domains appear to have a preferential ligand-interaction site distinct from the carbohydrate-binding sites found in lectins, and this interaction site accommodates not only sugars but also steroids and proteins. In fact, the LG/LNS domain interaction site has features reminiscent of the antigen combining sites in immunoglobulins. The LG/LNS domain presents an interesting case in which the fold has remained conserved but the functional sites have evolved; consequently, making predictions of structure-function relationships on the basis of the lectin fold alone is difficult. PMID- 11406410 TI - Histidine kinases and response regulator proteins in two-component signaling systems. AB - Phosphotransfer-mediated signaling pathways allow cells to sense and respond to environmental stimuli. Autophosphorylating histidine protein kinases provide phosphoryl groups for response regulator proteins which, in turn, function as molecular switches that control diverse effector activities. Structural studies of proteins involved in two-component signaling systems have revealed a modular architecture with versatile conserved domains that are readily adapted to the specific needs of individual systems. PMID- 11406411 TI - RNA editing by base deamination: more enzymes, more targets, new mysteries. AB - The posttranscriptional modification of messenger RNA precursors (pre-mRNAs) by base deamination can profoundly alter the physiological function of the encoded proteins. The recent identification of tRNA-specific adenosine deaminases (ADATs) has led to the suggestion that these enzymes, as well as the cytidine and adenosine deaminases acting on pre-mRNAs (CDARs and ADARs), belong to a superfamily of RNA-dependent deaminases. This superfamily might have evolved from an ancient cytidine deaminase. This article reviews the reactions catalysed by these enzymes and discusses their evolutionary relationships. PMID- 11406412 TI - Nuclear-receptor interactions on DNA-response elements. AB - Nuclear receptors regulate transcription by binding to DNA-response elements using their conserved DNA-binding domains. These response elements contain conserved hexameric sequences that can be arranged in various bipartite configurations, including inverted and direct repeats. A series of structural studies on receptor--DNA binding complexes illustrate the strategies used by receptors to recognize the symmetry of their binding site as well as its sequence. These structures also indicate how cooperation between receptors enhances their joint affinity and selectivity for correctly configured sites. PMID- 11406413 TI - The mitochondrial apoptosome: a killer unleashed by the cytochrome seas. AB - The caspase family of cysteine proteases have emerged as central regulators of apoptosis. Diverse cellular stresses trigger caspase activation by promoting release of mitochondrial components, including cytochrome c, into the cytoplasm. In turn, cytochrome c promotes the assembly of a caspase-activating complex termed the apoptosome. In this article, the apoptosome and its role in life and death decisions of cells are discussed. PMID- 11406414 TI - Xeroderma pigmentosum: the first of the cellular caretakers. PMID- 11406416 TI - Characterization of a single nucleotide polymorphism in the coding sequence of the bovine transferrin gene. AB - A single nucleotide polymorphism was identified in the coding sequence of the bovine transferrin gene. Two alleles (SSCP1 and SSCP2) were detected by SSCP analysis and the mutation point was identified and confirmed by direct sequencing of the PCR products. The relationship between protein and DNA polymorphism was established. Protein variants A, D1 and E correspond to SSCP allele 1 and variant D2 corresponds to SSCP allele 2. DNA sequences from genotypes AA, AE, AD2, D1E, D2E and D2D2 reveal an A/G substitution at position 1455 of the cDNA which causes a Gly/Glu substitution which could be responsible for the mobility difference between D1 and D2 variants. Because of the number of variants, this suggests that other SNPs exist in the bovine transferrin gene. A linkage analysis between the SSCPs and two microsatellites (UWCA46 and CSSM019) mapped the transferrin gene to BTA1. Two-point analysis revealed a tight linkage within the transferrin protein variants and the SSCPs. PMID- 11406417 TI - VAPSE-based analysis: a two-phased candidate gene approach for elucidating genetic predisposition to complex disorders. AB - The extraordinary success of linkage analysis in diseases with Mendelian inheritance has not extended readily to the genetics of common complex diseases. VAPSE-based analysis is a type of candidate gene approach that represents an alternative strategy by which genetic mechanisms can be defined despite the presence of substantial genetic heterogeneity. Recent advances in mutation screening and statistical methodology have enhanced substantially the efficiency and power of this approach. The "bread and butter" of VAPSE-based analysis is genotype-to-phenotype searches in large populations with computerized medical records. PMID- 11406418 TI - Genomic analysis of gamma-ray-induced germ-cell mutations at the b locus recovered from the medaka specific-locus test. AB - To study how gamma-ray-induced germ-cell mutations are fixed at the early embryonic stage of the next generation, genomic alterations in the b locus mutants (colorless melanophores) detected during development in the medaka specific-locus test (SLT) were analyzed. First, nine anonymous DNA markers linked to the b locus were cloned and mapped into the region extending about 47cM surrounding the b locus. Next, losses of paternal alleles of these DNA markers were examined in each of the 51 gamma-ray-induced b locus mutants obtained after irradiation of sperm or spermatids. In these mutants, 47 were dominant lethals, three were semi-viable and one was viable. All the mutants examined had large deletions surrounding the b locus. One viable mutant had an interstitial deletion, while all the semi-viable and dominant lethal ones appeared to have terminal deletions. Deletions extending about 20-35cM were the most frequently observed in 18 of the 51 mutants examined. The largest one extended more than 40cM. These results suggest that most of the gamma-ray induced germ cell mutations recovered as total specific-locus mutants were accompanied by large genomic deletions, which eventually led the mutant embryos to dominant lethality. PMID- 11406419 TI - Phylogenetic analysis of mitochondrial DNA in patients with an occipital stroke. Evaluation of mutations by using sequence data on the entire coding region. AB - Mitochondrial DNA (mtDNA) haplogroup U, defined by the polymorphism 12308A>G, may constitute a risk factor for an occipital stroke in migraine. We therefore identified 14 patients with an occipital stroke and with 12308A>G. We determined complete mtDNA coding region sequence for the patients and for population controls by conformation sensitive gel electrophoresis (CSGE) and direct sequencing. Sequence information was used to construct a phylogenetic network of mtDNA haplogroups U and K, which was found to be composed of subclusters U2, U4, U5 and a new subcluster U7, as well as cluster K. Five patients with a migrainous stroke belonged to subcluster U5 (P=0.006; Fisher's exact test). Many unique mutations were found among the patients with an occipital stroke including two tRNA mutations that have previously been suggested to be pathogenic. Analysis of mtDNA sequences by CSGE and comparison of the sequences through phylogenetic analysis greatly enhances the identification of mtDNA clusters in population and detection of mtDNA mutations in patients. PMID- 11406420 TI - Polymorphisms of the CYP1A1 and GSTM1 genes in relation to individual susceptibility to lung carcinoma in Chinese population. AB - Cytochrome P450 1A1 (CYP1A1) and glutathione S-transferase M1 (GSTM1) metabolize tobacco-related carcinogens. To investigate the prevalence of CYP1A1 and GSTM1, and their association with increased risk of lung carcinoma in Chinese, allele specific PCR and multiplex PCR technique were employed to identify the genotypes of CYP1A1 and GSTM1 in a case-control study of 106 lung carcinoma patients with histopathological diagnosis and 106 matched controls free of malignancy in Jiangsu Province, China. Logistic regression analysis was performed to calculate the odds ratio (OR) and 95% confidence intervals (CI). The results showed that individuals with GSTM1 null, and the combined GSTM1 null/CYP1A1 Ile/Val or GSTM1 null/CYP1A1 Val/Val had an elevated risk of lung carcinoma, with the OR, 1.92 (P=0.02; CI, 1.07-3.46), 3.27 (P=0.01; CI, 1.23-8.84) and 9.33 (P=0.04; CI, 1.01 217.42), respectively. Light smokers (<30 pack-years) carrying GSTM1 null genotype were shown to have the increased risk to lung carcinoma (OR=3.47; CI, 1.13-7.57). Our study suggested that the null GSTM1 genotype, independently or in combined with at least one Val allele of CYP1A1, might affect the genetic susceptibility for lung carcinoma in Chinese population. PMID- 11406421 TI - Polymorphic methyl group metabolism genes in patients with transitional cell carcinoma of the urinary bladder. AB - Because polymorphisms in the methyl group metabolism genes methylene tetrahydrofolate reductase (MTHFR), methionine synthase (MS), and cystathione beta-synthetase (CBS) affect plasma homocysteine levels and intracellular concentrations of S-adenosylmethionine (SAM), they modify the susceptibility to cardiovascular diseases and cancer. Specifically, genome-wide decreased DNA methylation ('hypomethylation') in human cancers might be a consequence of decreased SAM levels. Because hypomethylation is particularly prevalent in transitional cell carcinoma of the urinary bladder (TCC), the genotype distributions for the two each most prevalent MTHFR, MS, and CBS alleles were compared between 165 TCC patients and 150 population controls. The distributions of the MTHFR 677A/V and the MS 919G/D alleles were not significantly different between cancer patients and controls, even after stratification according to age, gender, tumor stage or grade. The CBS 844INS68 allele was slightly less frequent in TCC patients than in controls (q=0.07 versus 0.10), but was rarer among males in both groups. Among the TCC patients, this gender difference was highly significant (Mantel-Haenszel and chi(2)-test P=0.007). No significant difference between TCC patients and controls was found for any combined genotype. Likewise, the extent of DNA hypomethylation determined in 62 carcinoma specimens was not related to the respective genotypes. Thus, on their own, the MTHFR, MS and CBS genotypes do not appear to act upon susceptibility to TCC or influence the extent of DNA hypomethylation in this cancer. PMID- 11406422 TI - Evaluating the human optical system: corneal topography and wavefront analysis. AB - In this article we review three different systems for evaluating the human optical system: placido-disc based corneal topography, scanning slit topography, and wavefront sensors. We briefly describe the principles of each system, and discuss the strengths and weakness. We suspect all three will be used by ophthalmologists in the coming decade. PMID- 11406423 TI - Excimer laser photorefractive keratectomy. AB - The excimer laser has proved to be a precise and versatile tool for treating refractive errors. Excimer laser photorefractive keratectomy for myopia is now an established safe and effective alternative to contact lenses and glasses, and should be part of the ophthalmologist's standard armamentarium for treating this condition. This article provides a brief overview of the basic principles underlying excimer laser technology, as well as the surgical technique and postoperative management of myopic correction. PMID- 11406424 TI - Laser in situ keratomileusis: surgical technique. AB - Laser in-situ keratomileusis is an increasingly popular technique for the correction of refractive errors that was initially described by Pallikaris in 1990. It involves the excimer laser ablation of corneal stroma beneath a hinged corneal flap that is created with a microkeratome. The purpose of this chapter is to report the LASIK surgical technique that has evolved at the Emory Vision Correction Center since 1995 when an investigation study of this technique for the correction of myopia began. PMID- 11406425 TI - The intrastromal corneal ring segments: Intacs. AB - Although evolution of the intrastromal corneal ring technology will unquestionably continue, the ICRS in its current design is remarkably safe and offers several potential advantages, including preservation of the central cornea, reversibility and adjustability. The approval of this technology by the FDA has added a potentially unique procedure to the refractive surgeon's surgical inventory. PMID- 11406426 TI - Surgical correction of hyperopia: clear lens extraction and laser correction. AB - Modern ophthalmology offers a series of surgical procedures to correct a wide range of hyperopia and hyperopic astigmatism. Varies excimer lasers and thermal lasers have been proven safe and effective. Phakic implants and clear lens extraction offers alternatives for moderate to high hyperopic patients. Indications, techniques, and clinical study results were reviewed and summarized. PMID- 11406427 TI - Theory behind surgical correction of presbyopia. AB - This relatively new procedure is interesting, promising, and appears to be safe. These issues are currently being evaluated by the US FDA and in several other countries, most notably Canada. The procedure is based on Ronald A. Schachar's new theory of how accommodation occurs. The clinical course of the patients and the new wavefront analysis appear to support the theory. Regardless of the exact mechanism, clinically, the patients read and behave as though they have had their accommodation restored. The author certainly has been very impressed and excited with the results in his patients whose surgeons he has proctored. PMID- 11406428 TI - Phakic intraocular lenses. AB - Because of the potential risks associated with phakic IOL implantation, the current use of these devices is mainly in the treatment of high and extreme myopia which are expected to respond poorly to keratorefractive procedures. Overall, in this setting, the outcomes of phakic IOL implantation have been rather favorable, with significant improvement in uncorrected visual acuity, and tolerable visual symptoms in terms of glare and halo. PMID- 11406429 TI - Post implantation adjustable intraocular lenses. AB - To eliminate persistent refractive errors after cataract and phakic IOL surgery, photosensitive silicone IOLs have been developed. These IOL formulations enable precise laser adjustment of IOL power to correct spherical and toric errors post operatively, after wound and IOL stabilization. Initial experience with these laser adjustable IOLs indicate excellent biocompatability and adjustability of more than five diopters. PMID- 11406430 TI - Ultra-short pulse (femtosecond) laser surgery: initial use in LASIK flap creation. AB - The highly localized tissue effects of low energy femtosecond duration (ultrashort) laser pulses may be used to create three-dimensional intrastromal resections with micron precision and minimized collateral tissue damage. A surgical laser system that produces and delivers such pulses has been developed and tested clinically for creation of a corneal flap in LASIK. Expanded evaluation of this technology in this and additional keratorefractive applications is currently underway. PMID- 11406431 TI - Postoperative complications of excimer laser photorefractive keratectomy for myopia. AB - Excimer laser photorefractive keratectomy has become an increasingly popular surgical treatment for the correction of myopia. Unfortunately, no surgical technique can be entirely without complications. This article discusses the problems encountered following PRK for myopia in two main sections: those primarily refractive or optical in nature and those causing medical problems. PMID- 11406432 TI - Refractive surgery in the new millennium. AB - As we stand at the threshold of a new millennium, perhaps nowhere else in the field of ophthalmology is there such a feeling of excitement, enthusiasm, and anticipation as in the subspecialty of refractive surgery. What was once considered an experimental hobby, dabbled in by a few rogue physicians, has now gained a level of respect among patients and ophthalmologists that has surprised even its most vocal critics. PMID- 11406433 TI - The primary care physician and palliative care. AB - Palliative care is about total patient care, management of disabling and debilitating symptoms, and the role of the primary care physician. As physicians, we must understand who, what, when, how, and where to institute, or refer patients for palliative care. Who? Any patient needing assistance with the control of physical and nonphysical symptoms can benefit from palliative care. What? An interdisciplinary team coordinating care planning of the patient. When? The earlier in the disease process the patient is referred, the sooner symptom control can be maximally achieved. How? The primary care physician makes the initial referral to the palliative care physician or team. Where? All hospices should have a palliative care physician or consultant. This article provides a broad overview of palliative care management and incurable illnesses. PMID- 11406434 TI - Breaking bad news and discussing death. AB - The ability to discuss bad news with a patient and family is one clinical skill that is essential to providing effective end-of-life care. Patients and families value direct, nontechnical explanations that are given by a physician with compassion and kindness. Patients and families also value time to talk, express their feelings and ask questions. The authors review research on delivering bad news, then describe a six step process to guide physicians in discussing bad news with patients: (1) create an appropriate environment; (2) open the meeting; (3) discuss the news; (4) develop a follow-up plan; (5) document the conference; and (6) engage in self-reflection. PMID- 11406435 TI - Pain assessment and management. AB - Assessment and management of pain is crucial to the success of any program of care for dying patients and their families. With appropriate assessment and management, often using home health or hospice teams, pain can be controlled in more than 90% of patients. This article focuses on the symptomatic care of patients who are dying. The legal and regulatory issues that may inhibit delivery of adequate opioid therapy are also reviewed. PMID- 11406436 TI - Nonpain symptom management. AB - Excellent symptom management is paramount in palliative care. Without it, the individual patient will be unable to focus appropriately on other issues of concern, including those of a psychosocial, emotional, or spiritual nature. This article reviews current pharmacologic and nonpharmacologic interventions for symptoms commonly encountered in palliative care. These symptoms are organized into gastrointestinal, respiratory and neuropsychiatric categories. PMID- 11406437 TI - Management of emergent conditions in palliative care. AB - Emergent conditions arising in patients with advanced terminal disease are inevitable and create stressful situations for patients, caregivers, and health care personnel. Discussions regarding appropriate levels of intervention based on parameters such as patient wishes, proper access to documentation of those wishes, location of care, cognitive status and extent of clinical decline are important to have before crisis situations. Common emergencies to be addressed include those that may or may not be associated with advanced malignancies, including compression syndromes, superior vena cava syndrome, hypercalcemia, acute dyspnea, seizures, acute urinary and bowel obstructions, massive hemorrhage, cardiac tamponade, acute embolic phenomenon, and psychiatric emergencies. Although not all clinical scenarios will be addressed in this article, the more common ones will be discussed. PMID- 11406438 TI - The process of dying and managing the death event. AB - The end of life is a period of intensive care. It is imperative that physicians have training and skills in the care of dying patients. Understanding the process of dying and recognizing physical changes aids in prognostication. Skills in facilitating family coping, treating pain, respiratory distress and delirium are critical. PMID- 11406439 TI - Chronic illnesses and the end of life. AB - Great progress has been made in controlling the symptom distress of dying patients. This article reviews some of these methods and examines some triggering events that should cause physicians to re-examine patients' goals and care. PMID- 11406440 TI - Advance directives and establishing the goals of care. AB - This article addresses advance directives and methods to establish the goals of care for remaining life in the decisional and nondecisional patient. Without these discussions, patient autonomy is negated, and the opportunity to provide patient-centered care using shared decision making is lost. PMID- 11406441 TI - Pediatric palliative care. AB - This article presents a model of integrated palliative care for children with life-limiting illnesses, with emphasis on collaboration of care over time among family, primary care providers, and several other groups of providers. Some of the unique aspects of caring for children related to normal developmental changes and the family unit are considered. Issues related to pain and to specific diseases are also reviewed. PMID- 11406442 TI - Ethical and legal issues in palliative care. AB - As palliative care emerges as a respected and important component of contemporary health care, ethical issues will arise that confront and contest the provision of medical care. The basic principles of medical ethics, embodied in beneficence, nonmaleficence, autonomy, and justice, guide primary care physicians in dealing with dying patients. This article will discuss the basic ethical principles and the principle of double effect, decision-making capacity, advance directives, withholding and withdrawing life-sustaining therapy, futility, artificial nutrition and hydration, do-not-resuscitate orders, and physician-assisted suicide and euthanasia. PMID- 11406443 TI - Intercultural differences and communication at the end of life. AB - Cross-cultural encounters at the end of life are common and can result in misunderstandings and conflicts. The primary care physician is ideally suited to facilitate communication that can promote understanding and conflict resolution. PMID- 11406444 TI - Grief and bereavement. AB - Grief and bereavement are frequent concerns of primary care physicians. This article outlines grief in terminally ill patients and discusses interventions. Also included in this article is a review of the process of anticipatory grief and mourning and a discussion on normal and complicated grief. A portion of this article also covers grief in children and discusses interventions for grieving children. PMID- 11406445 TI - Hospice and palliative care programs. AB - Hospice and palliative care programs have grown rapidly in the United States over the last 25 years. Relief of suffering and maximization of quality of life including symptom control, psychosocial health, and spiritual care are the primary goals of hospice and palliative care. This article reviews the development, philosophy, and practice of hospice and palliative care, and describes barriers to and suggestions for integrating this approach into mainstream medicine. PMID- 11406446 TI - A surgeon's reflections on the care of the dying. AB - The characteristics, such as authority, power, certainty, and coolness under fire, that most surgeons consider essential to their ability to fight disease can be counterproductive in the care of the dying. Surgeons have begun to recognize that the ages-old image we have been taught to cherish must undergo some changes if we are to fulfill our responsibilities to those for whom our usual aggressive methods of treatment are to no avail. For this to happen, introspection and evaluation of the "inner surgeon" are required, in order to discover and deal with the hidden motivations that often influence decision-making at the bedside. The result of such self-analysis can be liberating. The new way of perceiving ourselves may bring us increased equanimity and break down some of the barriers that have stood in the way of effective palliative care. PMID- 11406447 TI - The surgeon and palliative care: an evolving perspective. AB - This article examines some of the cognitive and psychologic barriers surgeons possess when attempting to palliate advanced and incurable disease. Despite these barriers, the history of surgery demonstrates numerous eloquent exponents of palliative care philosophy. The considerable body of knowledge and experience in these matters beginning with the work of Dame Cicely Saunders and subsequent medical specialists is presented and offered as a guide for surgeons as their own perspective continues to evolve. PMID- 11406448 TI - The surgeon-patient relationship in advanced illness. AB - Surgeons are relatively new to palliative care, but there is a growing recognition of the contributions they can and should make. These go as much to the psychosocial support of the patient as they do to the technical aspects of their craft. The same qualities of proactivity and mastery of technique that stand the surgeon in good stead in the operating room can be acquired and mastered to make him or her equally effective in the hitherto nontraditional arenas of palliative care. PMID- 11406449 TI - Palliative philosophy: the ethical underpinning. AB - This article explores the ethical underpinnings of palliative care and how contemporary surgical care reinforces much of the basis of palliative medicine. A common misconception is that palliative medicine has grown out of the rise of patient's rights; however, palliative care is grounded in the principle of beneficence that is central to traditional medical values. This article explores the notion of beneficence to understand how contemporary surgical care can be undertaken and how it is consistent with palliative care. Some of the difficulties with the dichotomy between curative and palliative care models in surgical practice are discussed. Finally, this article considers how the underlying philosophy of palliative care is consistent with the historical tradition of surgery and the contemporary goals of surgical care. PMID- 11406450 TI - Spiritual dimensions of surgical palliative care. AB - Death is the most complex of religious issues and suicide is the most complex of philosophic issues. Our century has seen a massive technological expansion in our ability to prolong life, and unfortunately, to kill with unparalleled efficiency. Society has demanded that physicians explore with them whether death by suicide is a logical extension of palliative care. For surgeons, these profoundly spiritual concerns are, in many ways, beyond our education and training and may not be compatible with the current practice of surgery. PMID- 11406451 TI - The relationship between surgery and medicine in palliative care. AB - Palliative care represents the beneficence of surgery and medicine. A unified approach toward symptom control in patients with advanced diseases by surgeons and physicians is essential. Proposals are put forward to develop relationships between surgery and medicine for palliative care, teaching, and research. A model for integrating surgical and medical services to deliver the best palliative care is presented in this article. PMID- 11406452 TI - Pharmacologic management of pain: the surgeon's responsibility. AB - Historically, surgeons have had to witness their patients' pain probably longer than any specialty within medicine. Pain relief in palliative care forms the cornerstone of a comprehensive pattern of care that encompasses the physical, psychologic, social, and spiritual aspects of suffering. In a society that lives by mottoes, such as "no pain, no gain," and "just say no to drugs," pervasive subconscious barriers to effective pain relief exist. In being responsible for effective pain management to the patient, the surgeon must first set aside his or her own beliefs and attitudes regarding pain and its control and be open to change. PMID- 11406453 TI - Pharmacologic management of nonpain symptoms in surgical patients. AB - Palliative care patients present with multiple symptoms other than pain and cachexia. Asthenia, delirium, dyspnea, and chronic nausea and constipation cause significant distress to patients and families and frequently coexist in the same patient. A careful assessment frequently identifies reversible causes. When none are identified or there is no improvement, symptomatic pharmacologic interventions are available. The success rate is variable, and it is very high for symptoms such as chronic nausea or constipation and less effective for symptoms such as asthenia and delirium. More research on the mechanism and treatment of these symptoms is needed. PMID- 11406454 TI - The cancer cachexia syndrome. AB - The provision of additional calories and protein alone has not been shown to be efficacious in patients with cancer cachexia. Although primary research continues to unravel the complex metabolic derangements and diverse mediator pathways underlying cancer cachexia, the future lies in drugs and neutracenticals that may modulate this altered metabolism and enable conventional nutritional support to effectively replenish vital lean tissue. PMID- 11406455 TI - Invasive analgesia techniques for advanced cancer pain. AB - Because most cancer pain involves multiple anatomic sites, invasive techniques are intended to be analgesic adjuvants and not serve as the definitive treatment. These procedures often allow patients to reduce their dosages in their current drug regimens or to derive greater pain relief from their present doses in order to improve their quality of life. Medical care of the suffering pain patient requires a multimodality, multispecialty approach combining psychotherapy, social support, and pain management to provide the best possible quality of life or quality of dying. PMID- 11406456 TI - Critical palliative care: intensive care redefined. AB - In the area of end-of-life bioethical issues, patients, families, and health care providers do not understand basic principles, often leading to anguish, guilt, and anger. Providers lack communication skills, concepts, and practical bedside information. Linking societal values of the sanctity of life and quality of life with medical goals of preservation of life and alleviation of suffering respectively provides an essential structure. Medical care focuses on cure when possible but when the patient is dying, the focus switches to caring for patients and their families. Clinicians need to learn how to balance the benefits and burdens of medications and treatments, control symptoms, and orchestrate withdrawal of treatment. Finally, all need to learn more about the dying process to benefit society, their own families, and themselves. PMID- 11406457 TI - Palliative general surgical procedures. AB - Two types of procedure may be indicated in incurable patients. The first is palliative, in which the goal of intervention is relief of symptoms. The second type is supportive, where the procedure is a technical intervention done as part of a multidisciplinary treatment plan. The most minimally invasive but effective procedure is chosen. Procedures are categorized by the type of symptom the procedure is intended to relieve. This article emphasizes the principles involved in patient selection and outcome assessment in order to identify areas where more research is needed. PMID- 11406458 TI - The palliative uses of radiation therapy in surgical oncology patients. AB - Palliative radiation therapy has a significant role to play in the management of several clinical syndromes commonly seen by surgeons: brain and bone metastases, spinal cord compression, thoracic symptoms caused by lung cancer, bleeding, and pain or other local symptoms caused by tumor mass effects. This article discusses the basic principles of radiation therapy and radiobiology, the potential benefits, and the adverse effects and burdens of the treatment plans for each indication. This article gives the surgeon insight as to the proper time to refer patients with advanced cancer to a radiation oncologist for palliative radiation therapy. PMID- 11406459 TI - Palliative medical oncology. AB - Most advanced cancer patients remain incurable. They are carrying the tumor burden and the burden of symptoms as a result of physical, emotional, and spiritual distress. Modern medical oncology, sophisticated as it may appear in its endeavors to cure cancers, has historically failed to consider pain and symptom control as part of cancer care. Because of this, palliative medicine has emerged as the champion of pain and symptom control in advanced cancer patients. The reasons for medical oncology not embracing palliative care are analyzed and a model for palliative medical oncology is proposed. PMID- 11406460 TI - A perspective on physician-assisted dying. AB - This article discusses legalization of physician-assisted dying. Already much of the public is in favor of it, as are many physicians. Recent court decisions have so highlighted the diversity of thought on this issue that many people question whether there needs to be an absolute right or wrong. Patients who are dying slowly and painfully know that unnecessary suffering is being forced on them by conservative elements in our society. They feel that their desire for relief and for greater dignity and autonomy should receive more respect. PMID- 11406461 TI - Utility of genetic approaches to common cardiovascular diseases. PMID- 11406462 TI - Cytochalasin D induces edema formation and lowering of interstitial fluid pressure in rat dermis. AB - The increased capillary fluid filtration required to create a rapid edema formation in acute inflammation can be generated by lowering the interstitial fluid pressure (P(IF)). The lowering of P(IF) appears to involve dynamic beta(1) integrin-mediated interactions between dermal cells and extracellular matrix fibers. The present study specifically investigates the role of the cell cytoskeleton, i.e., the contractile apparatus of cells, in controlling P(IF) in rat skin as the integrins are linked to both the cytoskeleton and the extracellular matrix. P(IF) was measured using a micropuncture technique in the dorsal skin of the hind paw at a depth of 0.2--0.5 mm and following the induction of circulatory arrest with the intravenous injection of KCl in pentobarbital anesthesia. This procedure prevented the transcapillary flux of fluid and protein leading to edema formation in acute inflammation, which in turn can increase the P(IF) and therefore potentially mask a decrease of P(IF). Control P(IF) (n = 42) averaged -0.8 +/- 0.5 (means +/- SD) mmHg. In the first group of experiments, subdermal injection of 2 microl cytochalasin D, a microfilament-disrupting drug, lowered P(IF) to an average of -2.8 +/- 0.7 mmHg within 40 min postinjection (P < 0.05 compared with control). Subdermal injection of vehicle (10% DMSO in PBS or PBS alone) did not change the P(IF) (P > 0.05). Lowering of the P(IF) was not observed after the injection of colchicine or nocodazole, which specifically disrupts microtubuli in cultured cells. In the second group of experiments, 2 microl of cytochalasin D injected subdermally into rats with intact circulation increased the total tissue water (TTW) and albumin extravasation rate (E(ALB)) by 0.7 +/- 0.2 and 0.4 +/- 0.3 ml/g dry wt, respectively (P < 0.05 compared with vehicle). Nocodazole and colchicine did not significantly alter the TTW or E(ALB) compared with the vehicle (P > 0.05). Taken together, these findings strongly suggest that the connective tissue cells can participate in control of P(IF) via the actin filament system. In addition, the observation that subdermal injection of cytochalasin D lowered P(IF) indicates that a dynamic assembly and disassembly of actin filaments also occurs in the cells of dermal tissues in vivo. PMID- 11406463 TI - Catecholamines stimulate interleukin-6 synthesis in rat cardiac fibroblasts. AB - Proinflammatory cytokines have been implicated in the pathophysiology of different heart diseases. Recent evidence suggests that interleukin-6 (IL--6) may play a role in mechanisms leading to cardiac hypertrophy. In addition, catecholamines are known to induce cardiac hypertrophy. In the present study, we examined whether cardiac fibroblasts may be a potential source of IL--6 production in the rat heart and whether catecholamines can modulate the IL--6 synthesis. Only a small amount of IL--6 mRNA was detected in unstimulated rat cardiac fibroblasts. However, a 50-fold increase of IL--6 mRNA was found after stimulation with norepinephrine (NE). Addition of carvedilol, a alpha- and beta adrenergic receptor antagonist, prevented almost completely the NE-induced synthesis of IL--6 mRNA. Phenylephrine, an alpha-adrenergic agonist, and isoproterenol, a beta-adrenergic agonist, also induced an increase in IL--6. However, the stimulation via beta-receptors led to a more pronounced elevation. These data show that NE increases IL--6 expression in rat cardiac fibroblasts and that IL--6 may play an important autocrine/paracrine role in cardiac disease states associated with hypertrophy. PMID- 11406464 TI - Mechanism of temporal gradients in shear-induced ERK1/2 activation and proliferation in endothelial cells. AB - The aim of the current study was to investigate the intracellular signaling cascade that leads to temporal gradients in shear (TGS)-induced endothelial cell proliferation, with a focus on the involvement of extracellular signal-regulated kinases 1 and 2 (ERK1/2). With the use of well-defined pulsatile, impulse, step, and ramp laminar flow profiles, we found that TGS (impulse flow and pulsatile flow) induced an enhanced and sustained (>30 min) phosphorylation of ERK1/2 relative to step flow (which contains a step increase in shear followed by steady shear), whereas steady shear (ramp flow) alone downregulated activated ERK1/2. Nitric oxide (NO) was found to mediate both the stimulatory effect of TGS and the inhibitory effect of steady shear on endothelial ERK1/2 phosphorylation. Reactive oxygen species (ROS) were also demonstrated to be associated with TGS-induced ERK1/2 phosphorylation. Both G(q/11) and G(i3) were necessary for the activation of ERK1/2 by TGS. Finally, the TGS-induced endothelial proliferative response was abolished by ERK1/2 inhibition. Our study demonstrated the essential role of G proteins, NO, and ROS in TGS-dependent ERK1/2 activation and proliferative response in vascular endothelial cells. PMID- 11406465 TI - Differential ANG II-induced growth activation pathways in mesenteric artery smooth muscle cells from SHR. AB - Angiotensin II-induced growth signaling mechanisms were investigated in vascular smooth muscle cells (VSMCs) from mesenteric arteries of spontaneously hypertensive (SHR) and Wistar-Kyoto rats (WKY). In WKY, angiotensin II significantly increased protein synthesis ([(3)H]leucine incorporation) but not DNA synthesis ([(3)H]thymidine incorporation). In SHR, angiotensin II increased protein and DNA synthesis. VSMCs from both strains expressed angiotensin type 1 (AT(1)) and type 2 (AT(2)) receptors. Losartan (an AT(1) receptor antagonist) but not PD-123319 (an AT(2) receptor antagonist) attenuated angiotensin II-stimulated protein synthesis in WKY VSMCs. In SHR, losartan and PD-123319 partially inhibited angiotensin II-induced VSMC proliferation. The mitogen-activated protein kinase or extracellular signal-regulated protein kinase (ERK) kinase inhibitor PD-98059 blocked VSMC growth responses to angiotensin II in both strains. Angiotensin II increased ERK1/2 activation more in SHR than WKY, an effect inhibited by losartan but not PD-123319. LY-294002 [a phosphatidylinositol 3 (PI3) kinase inhibitor] blocked angiotensin II-stimulated ERK1/2 activation in SHR but not in WKY, whereas bisindolylmaleimide [a protein kinase C (PKC) inhibitor] was ineffective. In conclusion, angiotensin II stimulates VSMC proliferation via AT(1) and AT(2) receptors in SHR. In WKY, angiotensin II induces VSMC hypertrophy via AT(1) receptors. ERK1/2-dependent pathways regulated by intracellular Ca(2+) but not PKC mediate these effects. In SHR VSMCs, PI3 kinase plays a role in augmented angiotensin II-induced ERK1/2 phosphorylation. These angiotensin II-mediated signaling events could contribute to vascular remodeling in SHR. PMID- 11406466 TI - Inducible HSP70 mediates delayed cardioprotection via U-50488H pretreatment in rat ventricular myocytes. AB - To test the hypothesis that heat-shock proteins (HSPs) mediate delayed cardioprotection of prior kappa-opioid receptor (kappa-OR) stimulation, we first correlated cellular injury and viability with the expression of HSP70s in isolated rat ventricular myocytes subjected to prior kappa-OR stimulation with the selective agonist trans-(+/-)-3,4-dichloro-N-methyl-N-[2-(1 pyrrolidinyl)cyclohexyl]benzeneacetamide (U-50488H) and delayed lethal simulated ischemia (LSI). Cell injury and viability were indicated by lactate dehydrogenase release and trypan blue exclusion, respectively. The reduced injury and increased viability after pretreatment with U-50488H were concentration dependent and correlated directly with the expression of both stress-inducible (HSP70) and constitutive (HSC70) proteins. The effects mimic those with metabolic inhibition preconditioning (MIP). The cardioprotection against LSI by pretreatment with U 50488H and MIP was abolished and antagonized, respectively, via blockade of the kappa-OR by its selective antagonist, nor-binaltorphimine. We also found that blockade of the production of HSP70 but not HSC70 blocked the inhibitory effect of pretreatment with U-50488H on injury and viability. These observations provide evidence that stress-inducible HSP70 mediates delayed cardioprotection of prior kappa-OR stimulation. PMID- 11406467 TI - Cardioprotective actions of endogenous IL-10 are independent of iNOS. AB - Myocardial ischemia-reperfusion (I/R) is a well-known stimulus for acute inflammatory responses that promote cell death and impair pump function. Interleukin-10 (IL-10) is an endogenous, potent anti-inflammatory cytokine. Recently, it has been proposed that IL-10 inhibits inducible nitric oxide synthase (iNOS) activity after myocardial I/R and consequently exerts cardioprotective effects. However, whether this actually occurs remains unclear. To test this hypothesis, we utilized iNOS-deficient (-/-), IL-10 -/-, and IL 10/iNOS -/- mice to examine the potential mechanism of IL-10-mediated cardioprotection after myocardial I/R. Wild-type, iNOS -/-, IL-10 -/-, and IL 10/iNOS -/- mice were subjected to in vivo myocardial ischemia (30 min) and reperfusion (24 h). Deficiency of iNOS alone did not significantly alter the extent of myocardial necrosis compared with wild-type mice. We found that deficiency of IL-10 resulted in a significantly (P < 0.05) larger infarct size than that in wild-type hearts. Interestingly, deficiency of both IL-10 and iNOS yielded significantly (P < 0.01) larger myocardial infarct sizes compared with wild-type animals. Histological examination of myocardial tissue samples revealed augmented neutrophil infiltration into the I/R myocardium of IL-10 -/- and IL 10/iNOS -/- mice compared with hearts of wild-type mice. These results demonstrate that 1) deficiency of endogenous IL-10 exacerbates myocardial injury after I/R; 2) the cardioprotective effects of IL-10 are not dependent on the presence or absence of iNOS; and 3) deficiency of IL-10 enhances the infiltration of neutrophils into the myocardium after I/R. PMID- 11406468 TI - Effect of altered flow on the pattern of permeability around rabbit aortic branches. AB - Uptake of circulating macromolecules by the aortic wall is greater downstream than upstream of branch sites in immature rabbits, but the opposite pattern is seen at later ages. The mature pattern is nitric oxide dependent; we tested whether it is also flow dependent. Intercostal arteries of anesthetized rabbits were occluded, sham operated, or left alone. Uptake of rhodamine-labeled albumin was assessed by quantitative fluorescence microscopy of the sections through the aorta. In mature animals, uptake was higher upstream than downstream of the control and sham-operated branches, but the pattern was reversed at occluded branches. In young animals, uptake was not significantly different between regions upstream and downstream of control, sham-operated, or occluded branches. The absence of the normal immature pattern may reflect an influence of anesthesia and will assist in the elucidation of mechanisms underlying this pattern. The data for mature animals provide the first direct evidence that flow determines permeability near arterial branches and may account for the inverse spatial correlation between shear stress and disease prevalence at branches of adult human arteries. PMID- 11406469 TI - Parathyroid hormone-related protein-(1-34) inhibits intrinsic pump activity of isolated murine lymph vessels. AB - Parathyroid hormone-related protein (PTHrP) was originally found as a tumor derived vasoactive factor and has also been known to produce significant relaxation of vascular smooth muscles. Thus effects of PTHrP-(1-34), a PTH receptor-binding domain, on spontaneous lymphatic pump activity was investigated in isolated pressurized lymph vessels of mice. Low concentrations (1 x 10(-10) and 3 x 10(-10) M) of PTHrP-(1-34) dilated lymph vessels and reduced the frequency of pump activity, whereas high concentrations (1 x 10(-9) to 1 x 10(-8) M) of PTHrP-(1-34) caused dilation with cessation of the lymphatic pump activity. N(omega)-nitro-L-arginine methyl ester (L-NAME; 3 x 10(-5) M) but not indomethacin (1 x 10(-5) M) significantly reduced the PTHrP-(1-34)-induced inhibitory responses of the lymphatic pump activity. In the presence of L-NAME (3 x 10(-5) M) and L-arginine (1 x 10(-3) M), the L-NAME-induced inhibition in the PTHrP-(1-34)-mediated responses was significantly reduced. Glibenclamide (1 x 10( 6) M) significantly suppressed the inhibitory responses of the lymphatic pump activity induced by PTHrP-(1-34) and S-nitroso-N-acetyl-penicillamine. The PTHrP (1-34)-mediated inhibitory responses were significantly reduced by treatment with PTHrP-(7-34) (1 x 10(-7) M). These results suggest that PTHrP-(1-34) inhibits spontaneous pump activity of the isolated lymph vessels via PTH receptors and that production and release of endogenous nitric oxide and activation of ATP sensitive K(+) channels in the lymph vessels contribute to the PTHrP-(1-34) mediated inhibitory responses of the lymphatic pump activity. PMID- 11406470 TI - Selective A(2A) adenosine receptor activation reduces skin pressure ulcer formation and inflammation. AB - Activation of A(2A) adenosine receptors (A(2A)-AR) by ATL-146e (formerly DWH 146e) prevents inflammatory cell activation and adhesion. Recurrent ischemia reperfusion (I/R) of the skin results in pressure ulcer formation, a major clinical problem. ATL-146e was evaluated in a novel reproducible rat model of pressure ulcer. A 9-cm(2) region of dorsal rat skin was cyclically compressed at 50 mmHg using a surgically implanted metal plate and an overlying magnet to generate reproducible tissue necrosis. Osmotic minipumps were implanted into 24 rats divided into four equal groups to infuse vehicle (control), ATL-146e (0.004 microg x kg(-1) x min(-1)), ATL-146e plus an equimolar concentration of A(2A) antagonist, ZM-241385, or ZM-241385 alone. Each group received 10 I/R cycles. In non-I/R-treated skin, ATL-146e has no effect on blood flow. I/R-treated skin of the ATL-146e group compared with the vehicle group had 65% less necrotic area, 31% less inhibition of average skin blood flow, and fewer extravasated leukocytes (23 +/- 3 vs. 49 +/- 6 per 500 microm(2)). These data suggest that ATL-146e, acting via an A(2A)-AR, reduces leukocyte infiltration and is a potent prophylactic for I/R injury in skin. PMID- 11406471 TI - HMG-CoA reductase inhibitor stabilizes rabbit atheroma by increasing basal NO and decreasing superoxide. AB - Male rabbits fed a 0.5% cholesterol diet for 8 wk were divided into three groups. Group 1 was hypercholesterolemic; group 2 was fed a regular diet for an additional 12 wk; and group 3 was fed a regular diet with simvastatin (5 mg x kg( 1) x day(-1)). Simvastatin treatment reduced the atherosclerotic area and total and esterified cholesterol concentrations in the thoracic aorta. Tone-related basal nitric oxide (NO) release was highest in group 3. Acetylcholine-induced, NO dependent relaxation was improved in group 3 compared with group 2. Amount of endothelial nitric oxide synthase (eNOS) mRNA in vessels increased in group 1, compared with normal aorta, and decreased in group 2; however, it did not decrease in group 3. The amount of O released from vessels increased in group 1 and group 2 compared with normal rabbits; however, it decreased in group 3, especially in the endothelial cells. Peroxynitrite determined by nitrotyrosine staining decreased in group 3. Additionally, the arteries of rabbits fed a regular diet with or without simvastatin were investigated. The aorta from simvastatin-treated group showed increase of tone-related basal NO release and eNOS mRNA and decrease of O release. Taken together, upregulation of eNOS and decrease of O treatment were observed in vivo in the process of the sufficient stabilization of atheroma following simvastatin. PMID- 11406472 TI - Hemorrhage-induced alpha-adrenergic signaling results in myocardial TNF-alpha expression and contractile dysfunction. AB - Hemorrhagic shock (HS), secondary to major blood loss, frequently precedes multiple organ dysfunction and is accompanied by a surge in circulating catecholamine levels. Expression of the cardiodepressant cytokine, tumor necrosis factor-alpha (TNF-alpha), has been observed in the heart after HS and resuscitation (HS/R) and alpha(1)-adrenergic blockade prevented translocation of the nuclear transcription factor, NF-kappa B, to the nucleus. We hypothesized that alpha(1)-adrenergic stimulation induces myocardial TNF-alpha expression, which results in depressed cardiac function after HS/R. The role of alpha(1) adrenergic stimulation in myocardial TNF-alpha expression and depressed cardiac function after HS/R was assessed by treatment with the alpha(1)-adrenergic inhibitor, prazosin hydrochloride (1 mg/kg ip), for 1 h before the onset of hemorrhage. In addition, TNF-alpha was neutralized with a specific antibody (600 microl/kg iv) 5 min before hemorrhage. HS was induced by the withdrawal of blood to a mean blood pressure of 50 mmHg for 1 h. Contractile function was measured with the use of a Langendorff apparatus 2 h after the end of HS. HS/R led to significant decreases in left ventricular developed tension and in the maximal rate of pressure increase over time during both contraction and relaxation. Myocardial expression of TNF-alpha measured by enzyme-linked immunosorbent assay increased significantly after 30 min of hemorrhage and peaked after 60 min of HS and 45 min of resuscitation. Depression in cardiac function after HS/R was reversed by 85% in hearts from rats treated with a TNF-alpha neutralizing antibody and by 90% in hearts from rats treated with prazosin hydrochloride. We conclude that HS activates a alpha(1)-adrenergic pathway, resulting in TNF-alpha expression in the heart and depressed myocardial contractile function. PMID- 11406473 TI - Activation of cardiac afferents by arachidonic acid: relative contributions of metabolic pathways. AB - Arachidonic acid (AA) is metabolized via cyclooxygenase (COX), lipoxygenase (LOX), and cytochrome P-450 (CP450) pathways to a variety of bioactive products. The sensitivity of cardiac afferent endings to AA and its metabolites, especially those derived from LOX and CP450 pathways, is currently unclear. We examined AA induced activation of cardiac vagal chemosensitive afferents in non- and postischemic hearts in rats and evaluated the relative contributions of the three metabolic pathways to the effects. Epicardial application of AA activated the cardiac afferents dose dependently in both nonischemic and postischemic hearts, with afferent responses greater in the latter condition. In nonischemic hearts, the afferent response to AA was abolished only after simultaneous administration of indomethacin and 17-octadecynoic acid (COX and CP450 inhibitors, respectively). Nordihydroguaiaretic acid (a LOX inhibitor) had no effect on the afferent response to AA. In postischemic hearts, abolition of the afferent response to AA required simultaneous blockade of all three pathways. None of the AA metabolic inhibitors affected resting activity of cardiac afferents in nonischemic hearts, but each suppressed afferent activity during ischemia reperfusion. Most COX metabolites, CP450 metabolites, and 5-LOX metabolites tested were capable of activating cardiac afferents. The 12-LOX metabolites and 15-LOX metabolites had no effect on afferent activity. These data indicate that in the nonischemic heart, basal AA metabolism does not contribute to resting afferent activity, but AA is capable of activating cardiac afferents via COX and CP450 but not LOX pathways. During ischemia-reperfusion, all three metabolic pathways contribute to activation of cardiac vagal afferents with an enhanced responsiveness to AA. Our results suggest that induction of the 5-LOX pathway contributes to the enhanced sensitivity of cardiac vagal afferents to AA in the ischemic condition. PMID- 11406474 TI - Shear stress regulates occludin content and phosphorylation. AB - Previous studies determined that shear stress imposed on bovine aortic endothelial cell (BAEC) monolayers increased the hydraulic conductivity (L(P)); however, the mechanism by which shear stress increases L(P) remains unknown. This study tested the hypothesis that shear stress regulates paracellular transport by altering the expression and phosphorylation state of the tight junction protein occludin. The effect of shear stress on occludin content was examined by Western blot analysis. Ten dyn/cm(2) significantly reduced occludin content in a time dependent manner such that after a 3 h exposure to shear, occludin content decreased to 44% of control. Twenty dyn/cm(2) decreased occludin content to 50% of control and increased L(P) by 4.7-fold after 3 h. Occludin expression and L(P) depend on tyrosine kinase activity because erbstatin A (10 microM) attenuated both the shear-induced decrease in occludin content and increase in L(P). Shear stress increased occludin phosphorylation after 5 min, 15 min, and 3 h exposures. The shear-induced increase in occludin phosphorylation was attenuated with dibutyryl (DB) cAMP (1 mM), a reagent previously shown to reverse the shear induced increase in L(P). We conclude that shear stress rapidly (< or = 5 min) increases occludin phosphorylation and significantly decreases the expression of occludin over 1-4 h. Alterations in the occludin phosphorylation state and occludin total content are potential mechanisms by which shear stress increases L(P). PMID- 11406475 TI - Regulation of ERK phosphorylation in differentiated arterial muscle of rabbits. AB - Extracellular signal-regulated kinases (ERK) and mitogen-activated protein (MAP) kinases participate in cell signaling, regulating cell growth. In differentiated cells, the role ERK plays is less well known. This study quantified the degree of basal and stimulated ERK phosphorylation and contraction in freshly isolated arteries. The level of basal ERK phosphorylation was identical in preloaded and slack arteries, was greater in media than in the whole artery, and was reduced by the MAP or ERK kinase (MEK) inhibitor PD-98059. Chemical denudation using 1H [1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one did not elevate basal ERK phosphorylation. PD-98059 reduced maximum phenylephrine (PE)-stimulated ERK phosphorylation but not force. Pervanadate elevated ERK phosphorylation without causing contraction. Contractions produced by PE and relaxations produced by PE washout preceded the ERK phosphorylation. K(+) depolarization, muscle stretch, and angiotensin II elevated ERK phosphorylation transiently, whereas PE maintained ERK phosphorylation for 30 min. The alpha(1A)-adrenergic receptor antagonist WB-4101 reduced PE-stimulated force by 70% and abolished PE-induced ERK phosphorylation. Afterloaded and zero-load contractions produced by K(+) depolarization displayed identical increases in ERK phosphorylation. These data indicate that ERK was active basally in the differentiated artery but regulated by the endothelium and that ERK phosphorylation was not load dependent. A strong correlation between PE-induced force and ERK phosphorylation supports the hypothesis that ERK activation may reflect a signal "notifying" the cell of the degree of alpha(1)-adrenergic receptor-induced contraction. PMID- 11406476 TI - Endothelin-1 and nitric oxide synthase in short rebound reaction to short exposure to inhaled nitric oxide. AB - On withdrawal of inhalation of nitric oxide (INO) administered after lung injury, pulmonary artery pressure (PAP) and arterial oxygen tension (Pa(O(2))) may deteriorate more than before INO (rebound response). In this study, we investigated the possible roles of endothelin (ET)-1 and nitric oxide (NO) synthase (NOS) activity in the short rebound reaction to short-term inhalation of NO. Twenty-six anesthetized mechanically ventilated piglets were given endotoxin infusion. Twelve animals then received INO (30 parts per million) for two 30-min periods. Nine controls were not given NO. Measurements were made of blood gases and hemodynamic parameters, lung tissue ET-1 expression and NOS activity, and plasma ET-1 concentration. INO decreased PAP and increased Pa(O(2)), but INO withdrawal caused a short rebound reaction with an increase in PAP. Lung tissue expression and plasma concentration of ET-1 increased during INO, and plasma ET-1 increased further after its withdrawal. Activity of constitutive NOS decreased during INO, whereas that of inducible NOS was unchanged. Upregulation of ET-1 and downregulation of NOS activity may have influenced the short rebound reaction to short-term INO. PMID- 11406477 TI - Intermittent hypoxia modulates nNOS expression and heart rate response to sympathetic nerve stimulation. AB - Nitric oxide (NO) decreases norepinephrine (NE) release and the heart rate (HR) response to sympathetic nerve stimulation (SNS). We tested the hypothesis that the enhanced HR response to sympathetic activation following chronic intermittent hypoxia (IH) results from a peripheral modulation of pacemaking by NO. Isolated guinea pig double atrial/right stellate ganglion preparations were studied from animals that had been exposed to IH (n = 20) and control animals (n = 22). The HR response to SNS was significantly enhanced in the IH group compared with the controls. However, the increase in HR with cumulative doses (0.1--10 microM) of bath-applied NE was similar in both groups. Western blot analysis showed less neuronal NO synthase in the right atria from the IH group. In IH animals, the NO synthase inhibitor, N(omega)-nitro-L-arginine (L-NNA; 100 microM) did not alter the increased HR response to SNS, whereas in control animals L-NNA significantly increased the HR response to SNS; an effect that was reversed with excess L arginine. In conclusion, the enhanced HR response to SNS after IH may be related to a decreased inhibitory action of NO on presynaptic NE release. PMID- 11406478 TI - In vivo assessment of acetylcholine-releasing function at cardiac vagal nerve terminals. AB - We examined whether the ACh concentration measured by cardiac microdialysis provided information on left ventricular ACh levels under a variety of vagal stimulatory and modulatory conditions in anesthetized cats. Local administration of KCl (n = 5) and ouabain (n = 7) significantly increased the ACh concentration in the dialysate to 4.3 +/- 0.8 and 7.3 +/- 1.3 nmol/l, respectively, from the baseline value of 0.6 +/- 0.5 nmol/l. Intravenous administration of phenylbiguanide (n = 5) and phenylephrine (n = 6) significantly increased the ACh concentration to 5.4 +/- 0.9 and 6.0 +/- 1.5 nmol/l, respectively, suggesting that the Bezold-Jarisch and arterial baroreceptor reflexes affected myocardial ACh levels. Modulation of vagal nerve terminal function by local administration of tetrodotoxin (n = 6), hemicholinium-3 (n = 6), and vesamicol (n = 5) significantly suppressed the electrical stimulation-induced ACh release from 20.4 +/- 3.9 to 0.6 +/- 0.1, 7.2 +/- 1.9, and 2.7 +/- 0.6 nmol/l, respectively. Increasing the heart rate from 120 to 200 beats/min significantly reduced the myocardial ACh levels during electrical vagal stimulation, suggesting a heart rate-dependent washout of ACh. We conclude that ACh concentration measured by cardiac microdialysis provides information regarding ACh release and disposition under a variety of pathophysiological conditions in vivo. PMID- 11406479 TI - Differential effects of natriuretic peptides and NO on LV function in heart failure and normal dogs. AB - beta-Adrenergic hyporesponsiveness in congestive heart failure (CHF) is mediated, in part, by nitric oxide (NO). NO and brain natriuretic peptide (BNP) share cGMP as a second messenger. Left ventricular (LV) function and inotropic response to intravenous dobutamine (Dob) were assessed during sequential intracoronary infusion of saline, HS-142-1 (a BNP receptor antagonist), and HS-142-1 + N(G) monomethyl-L-arginine (L-NMMA) in anesthetized dogs with CHF due to rapid pacing and in normal dogs during intracoronary infusion of saline, exogenous BNP, and sodium nitroprusside (SNP). In CHF dogs, intracoronary HS-142-1 did not alter the inotropic response to Dob [percent change in first derivative of LV pressure (% Delta dP/dt) 47 +/- 4% saline vs. 54 +/- 7% HS-142-1, P = not significant]. Addition of intracoronary L-NMMA to HS-142-1 enhanced the response to Dob (% Delta dP/dt 73 +/- 8% L-NMMA + HS-142-1, P < 0.05 vs. H142-1). In normal dogs, intracoronary SNP blunted the inotropic response to Dob (% Delta dP/dt 93 +/- 6% saline vs. 71 +/- 5% SNP, P < 0.05), whereas intracoronary BNP had no effect. In CHF dogs, the time constant of LV pressure decay during isovolumic relaxation increased with intracoronary HS-142-1 (48 +/- 4 ms saline vs. 58 +/- 5 ms HS-142 1, P < 0.05) and further increased with intracoronary L-NMMA (56 +/- 6 ms HS-142 1 vs. 66 +/- 7 ms L-NMMA + HS-142-1, P < 0.05). Endogenous BNP and NO preserve diastolic function in CHF, whereas NO but not BNP inhibits beta-adrenergic responsiveness. PMID- 11406480 TI - Estrogen restores postischemic pial microvascular dilation. AB - Estrogen protects the brain from experimental cerebral ischemia, likely through both vascular and neuronal cellular mechanisms. The purpose of this study was to determine whether chronic estrogen treatment in males and repletion in ovariectomized (Ovx) females reverses abnormalities in pial arteriolar reactivity during reperfusion from global forebrain ischemia (4-vessel occlusion, 15 min) and whether the site of protection is vascular endothelium. Male and Ovx female rats were implanted with either placebo or a 25-microg 17 beta-estradiol pellet 10 days before ischemia. With the use of intravital microscopy, pial vessel dilation to ACh (10 microM) and S-nitroso-N-acetyl-penicillamine (SNAP; 1 microM) and vasoconstriction to serotonin (10 microM) was examined in situ at 30--60 min of reperfusion. Postischemic changes in vessel diameter were compared with preischemic values for each agent. Postischemic response to both ACh and SNAP was lost in males and Ovx females, but not in estrogen pellet-implanted males and estrogen-implanted Ovx females, suggesting that estrogen protects both endothelial and smooth muscle-mediated vasodilation. Ischemia blunted vessel constriction to serotonin regardless of treatment. These data demonstrate that estrogen acts as a vasoprotective agent within the cerebral circulation and can improve microvascular function under conditions of an acutely evolving ischemic pathology. PMID- 11406481 TI - Angiotensin II regulates phosphorylation of translation elongation factor-2 in cardiac myocytes. AB - Increased protein synthesis is the cardinal feature of cardiac hypertrophy. We have studied angiotensin II (ANG II)-dependent regulation of eukaryotic elongation factor-2 (eEF-2), an essential component of protein translation required for polypeptide elongation, in rat neonatal cardiac myocytes. eEF2 is fully active in its dephosphorylated state and is inhibited following phosphorylation by eEF2 kinase. ANG II treatment (10(-10) - 10(-7) M) for 30 min produced an AT(1) receptor-specific and concentration- and time-dependent reduction in the phosphorylation of eEF-2. Protein phosphatase 2A (PP2A) inhibitors okadaic acid and fostriecin, but not the PP2B inhibitor FK506, attenuated ANG II-dependent dephosphorylation of eEF-2. ANG II activated mitogen activated protein kinase, (MAPK) within 10 min of treatment, and blockade of MAPK activation with PD-98059 (1--20 nM) inhibited eEF-2 dephosphorylation. The effect of ANG II on eEF-2 dephosphorylation was also blocked by LY-29004 (1-20 nM), suggesting a role for phosphoinositide 3-kinase, but the mammalian target rapamycin inhibitor rapamycin (10--100 nM) had no effect. Together these results suggest that the ANG II-dependent increase in protein synthesis includes activation of eEF-2 via dephosphorylation by PP2A by a process that involves both PI3K and MAPK. PMID- 11406482 TI - Calcium-independent phospholipase A(2) mediates CREB phosphorylation and c-fos expression during ischemia. AB - In isolated, perfused adult rat hearts, global ischemia increased the phosphorylation of cAMP response element-binding protein (CREB) relative to control levels, and this phosphorylation was reversed with reperfusion. CREB phosphorylation elicited by 5 min of global ischemia was sensitive to treatments with the calcium-independent phospholipase A(2) (iPLA(2)) inhibitor bromoenol lactone (BEL) and occurred in the absence of increases in myocardial cAMP content. In contrast, CREB phosphorylation elicited by 15 min of global ischemia was likely mediated by elevated cAMP levels. The expression of c-fos, in response to brief myocardial ischemia, was also sensitive to BEL treatment. The induction of iPLA(2)-mediated CREB phosphorylation was further substantiated by the observations that lysoplasmenylcholine increased both the phosphorylation of CREB and the induction of c-fos expression in the absence and presence of BEL. CREB phosphorylation in both ischemic hearts and lysoplasmenylcholine-perfused hearts was inhibited by pretreatment of hearts with the specific cAMP-dependent protein kinase (PKA) inhibitor H-89. Taken together, these data demonstrate that iPLA(2) mediates CREB phosphorylation through a PKA-dependent pathway during brief periods of myocardial ischemia, possibly through the formation of lysophospholipids. PMID- 11406483 TI - Cross talk between prostacyclin and nitric oxide under shear in smooth muscle cell: role in monocyte adhesion. AB - We tested the hypothesis that at sites of vascular damage, vessel homeostasis is maintained through the cross talk of shear-induced production of prostacyclin and nitric oxide (NO) in vascular smooth muscle cells (VSMC). Confluent A7r5 cells derived from rat aortic VSMC and mesenteric VSMC were exposed to shear stress at 15 dyn/cm(2) for 90 min with the use of a cone-plate device, and productions of prostacyclin and NO were examined. Shear stress increased cumulative production of prostacyclin by 3- to 3.5-fold and that of NO by 6- to 7.5-fold. Western blot analysis showed that inducible NO synthase protein was expressed after shear stress in both types of VSMC. Inhibition of NO synthase enhanced the shear induced production of prostacyclin from 40 to 60%. Shear-induced production of NO was suppressed by 70% after treatment with 10(-4) M of indomethacin. A7r5 cells adhesiveness for monocytes was suppressed by 50% after shear stress. This suppression was abolished by pretreatment with 10(-4) M of indomethacin, whereas inhibition of NO synthase only minimally inhibited it. We conclude that there is a cross talk of shear-induced production of prostacyclin and NO in VSMC. At sites of vascular damage, prostacyclin synthesis may prevent monocyte adhesiveness for VSMC through the concomitant enhancement of NO production. PMID- 11406484 TI - Pregnancy enhances endothelium-dependent relaxation of ovine uterine artery: role of NO and intracellular Ca(2+). AB - The present study tested the hypothesis that the pregnancy-associated increase in endothelium-dependent relaxation of the uterine artery was mediated primarily by an increase in nitric oxide (NO) release, resulting in a reduction in smooth muscle intracellular Ca(2+) concentration ([Ca(2+)](i)). Uterine arteries obtained from nonpregnant and near-term (140 days gestation) pregnant sheep were used. The Ca(2+) ionophore A23187 induced endothelium-dependent relaxations in both nonpregnant and pregnant uterine arteries, with an increased relaxation in the pregnant tissue. In contrast, endothelium-independent relaxations induced by sodium nitroprusside were the same in nonpregnant and pregnant arteries. In addition, removal of the endothelium significantly increased noradrenaline induced contractions in pregnant, but not nonpregnant, uterine arteries. In accordance, pregnancy increased both basal and A23187-stimulated NO releases in the uterine artery. Simultaneous measurement of tension and [Ca(2+)](i) in the smooth muscle demonstrated a linear correlation with the slope of unity between A23187-induced relaxation and the reduction of [Ca(2+)](i) in both nonpregnant and pregnant uterine arteries. The A23187-induced reduction of [Ca(2+)](i) was significantly enhanced in pregnant, compared with nonpregnant, uterine arteries. The results indicate that pregnancy increases NO release, which, through decreasing [Ca(2+)](i) in the smooth muscle, accounts for the increased endothelium-dependent relaxation of the uterine artery. Signal transduction pathways distal to NO production are not changed by pregnancy. PMID- 11406485 TI - Role of nitric oxide and protein kinase C in ACh-induced cardioprotection. AB - We examined the roles of nitric oxide and protein kinase C (PKC) in ACh-produced protection of cultured cardiomyocytes during simulated ischemia and reoxygenation. Cell viability was quantified using propidium iodide in chick embryonic ventricular myocytes. O(2) radicals were quantified using 2',7' dichlorofluorescin diacetate. After a 10-min infusion of ACh (0.5 or 1 mM) and a 10-min drug-free period, we simulated ischemia for 1 h and reoxygenation for 3 h. ACh reduced cardiocyte death [32 +/- 4%; n = 6 and 23 +/- 4%; n = 7 (P < 0.05)] and attenuated oxidant stress during ischemia and reoxygenation in a concentration-dependent manner compared with controls (47 +/- 4%; n = 8; P < 0.05). The increase in O(2) radicals before simulated ischemia [357 +/- 49; n = 4 and 528 +/- 52; n = 8 vs. 211 +/- 34; n = 8; P < 0.05 (arbitrary units)] was abolished by the specific nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME) and was markedly attenuated by N(G)-monomethyl-L-arginine (L-NMMA). L-NAME or L-NMMA blocked the protective effects of ACh, which selectively increased PKC-epsilon isoform activity in the particulate fraction. The PKC inhibitor Go-6976 had no effect on O(2) radical production before simulated ischemia but it abolished the protection; therefore nitric oxide is a large component of ACh-generated O(2) radicals. Nitric oxide and O(2) radicals activate the PKC-epsilon isoform by which ACh protects against injury. PMID- 11406486 TI - Lack of role for nitric oxide in cholinergic modulation of myocardial contractility in vivo. AB - Despite intensive investigation, the role of nitric oxide (NO) in cholinergic modulation of myocardial contractility remains unresolved. The left anterior descending coronary artery of 34 anesthetized, open-chest dogs was perfused via an extracorporeal circuit. Segmental shortening (SS) was measured with ultrasonic crystals and coronary blood flow (CBF) was measured with an ultrasonic flow transducer. An intracoronary infusion of ACh (20 microg/min) was performed, with CBF held constant, under baseline and during dobutamine, CaCl(2), or amrinone at doses increasing SS by approximately 50% (10 microg/min, 15 mg/min, and 300 microg/min ic, respectively). ACh-induced responses during dobutamine were also assessed following treatment with the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME; 300 microg/min ic for 15 min). The effects of sodium nitroprusside (SNP; 80 microg/min ic), an exogenous NO donor, bradykinin (2.5 microg/min ic), a nonmuscarinic releaser of endothelial NO, and bilateral vagal stimulation (before and after L-NAME) were evaluated during dobutamine. ACh had no effect on SS under baseline or during CaCl(2), but it decreased SS during dobutamine or amrinone (-23 +/- 4% and -30 +/- 5%, respectively). Vagal stimulation also reduced SS during dobutamine. L-NAME did not alter the ACh- or vagal-induced decreases in SS during dobutamine. Neither SNP nor bradykinin affected SS during dobutamine. In conclusion, ACh and vagal stimulation have a negative inotropic effect during stimulation of the beta-adrenergic receptors that is independent of NO. The persistence of this effect during amrinone suggests that a mechanism downstream from adenylate cyclase is involved. PMID- 11406487 TI - Changes in collagenase and collagen gene expression after induction of aortocaval fistula in rats. AB - Progressive ventricular dilatation commonly accompanies the transition to overt failure in chronically overloaded hearts; however, only recently have studies begun to elucidate underlying molecular alterations. In particular, the potential role of altered myocardial expression of the procollagenase gene in this process has not previously been examined. Biventricular hypertrophy and dilatation were produced in rats by creating an abdominal aortocaval fistula. The time courses of changes in expression of collagen I and III genes and of the procollagenase gene (matrix metalloproteinase-1, MMP-1) were assessed by Northern blot hybridization. Expression of all three genes increased promptly; however, collagenase gene expression peaked much earlier (8 h) than did expression of either of the collagen genes (7 days), and all returned to baseline levels by 45 days. These data corroborate earlier reports of increased collagen gene expression in this model, but more importantly, they provide the first evidence of concurrent activation of collagenase gene expression, suggesting that enhancement of collagen degradation may be a prerequisite for structural cardiac dilatation. PMID- 11406488 TI - Protective effect of heat shock protein 72 on contractile function of perfused failing heart. AB - The contribution of heat shock protein 72 (HSP72) to the protection of cardiac function was examined in rats with chronic heat failure (CHF) following coronary artery ligation (CAL). The CAL animals revealed functional deterioration without low cardiac output 2 wk after CAL and with low cardiac output 8 wk after CAL, suggesting that CHF had developed by 8 wk after CAL. The hearts isolated from animals 2 and 8 wk after CAL (2-wk CAL heart and 8-wk CAL heart, respectively) were subjected to hyperthermia (at 42 degrees C) for 15 min, followed by 6-h perfusion (hyperthermia/6-h perfusion). The 2-wk CAL heart showed a 19.0 +/- 3.9% decline in the rate- pressure product (RPP) after hyperthermia/6-h perfusion, similar to the nonoperated control (19.8 +/- 2.9% decline). The production of myocardial HSP72 increased in the 2-wk CAL heart in response to hyperthermia (412.7 +/- 29.5% of each prehyperthermia value). The 8-wk CAL heart showed a reduction in the RPP (45.2 +/- 4.3% decline) after hyperthermia/6-h perfusion, associated with blunting of the production of HSP72 (68.9 +/- 22.6% increase, respectively). The results suggest that functional deterioration of the isolated failing heart may be attributed to a reduction in the production of myocardial HSP72. PMID- 11406489 TI - Sarcoplasmic reticulum Ca(2+) uptake is impaired in coronary smooth muscle distal to coronary occlusion. AB - After chronic occlusion, collateral-dependent coronary arteries exhibit alterations in both vasomotor reactivity and associated myoplasmic free Ca(2+) levels that are prevented by chronic exercise training. We tested the hypotheses that coronary occlusion diminishes Ca(2+) uptake by the sarcoplasmic reticulum (SR) and that exercise training would prevent impaired SR Ca(2+) uptake. Ameroid constrictors were surgically placed around the proximal left circumflex (LCx) artery of female swine 8 wk before initiating 16-wk sedentary (pen confined) or exercise-training (treadmill run) protocols. Twenty-four weeks after Ameroid placement, smooth muscles cells were enzymatically dissociated from both the LCx and nonoccluded left anterior descending (LAD) arteries of sedentary and exercise trained pigs, and myoplasmic free Ca(2+) was studied using fura 2 microfluorometry. After the SR Ca(2+) store was partially depleted with caffeine (5 mM), KCl-induced membrane depolarization produced a significant decrease in the time to half-maximal (t(1/2)) myoplasmic free Ca(2+) accumulation in LCx versus LAD cells of sedentary pigs. Furthermore, inhibition of sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA; 10 microM cyclopiazonic acid) significantly reduced t(1/2) in cells isolated from the LAD but not from the LCx. Exercise training did not prevent the differences in t(1/2) myoplasmic free Ca(2+) accumulation observed between LCx and LAD cells. Occlusion or exercise training did not alter SERCA protein levels. These results support our hypothesis of impaired SR Ca(2+) uptake in coronary smooth muscle cells isolated distal to chronic occlusion. Impaired SR Ca(2+) uptake was independent of SERCA protein levels and was not prevented by exercise training. PMID- 11406490 TI - Comparison of effects of diabetes mellitus on an EDHF-dependent and an EDHF independent artery. AB - The hypothesis tested in this study is that diabetes has a different impact on an artery in which endothelium-dependent responses derive from both nitric oxide (NO) and endothelium-derived hyperpolarizing factor (EDHF) compared with responses in which NO predominates and EDHF is absent. The streptozotocin-treated rat model of diabetes was used, and the arteries were mounted on a wire myograph. In mesenteric arteries depolarized and constricted with phenylephrine, acetylcholine evoked hyperpolarization (31 +/- 2 mV) and complete relaxation; these responses were attributed to EDHF and NO. In femoral arteries, acetylcholine evoked a small, NO-mediated hyperpolarization (5 +/- 1 mV) and incomplete relaxation. Bradykinin evoked NO-dependent responses in mesenteric arteries. Whereas diabetes significantly impaired the EDHF-dependent hyperpolarization and relaxation in mesenteric arteries, NO-dependent responses in femoral and mesenteric arteries were preserved. 1-Ethyl-2-benzimidazolinone evoked hyperpolarization and relaxation in mesenteric arteries, and this was impaired in diabetes. In conclusion, NO-dependent responses are preserved in diabetes, whereas endothelial responses-dependent upon EDHF appear to be impaired. The putative channels responsible for mediating the EDHF response may be altered in diabetes. PMID- 11406491 TI - Relationship between capillary angiogenesis, fiber type, and fiber size in chronic systemic hypoxia. AB - Whether chronic hypoxia causes angiogenesis in skeletal muscle is controversial. Male Wistar rats, 5--6 wk of age, were kept at constant 12% O(2) for 3 wk, and frozen sections of their postural soleus (SOL), phasic extensor digitorum longus (EDL), and tibialis anterior (TA) muscles were compared with those of normoxic controls. Capillary supply increased in SOL muscles [capillary-to-fiber ratio (C/F) = 2.55 +/- 0.09 hypoxia vs. 2.17 +/- 0.06 normoxia; capillary density (CD) = 942 +/- 14 hypoxia vs. 832 +/- 20 mm(-2) normoxia, P < 0.01] but not in EDL muscles (C/F = 1.44 +/- 0.04 hypoxia vs. 1.42 +/- 0.04 normoxia; CD = 876 +/- 52 hypoxia vs. 896 +/- 24 mm(-2) normoxia). The predominantly glycolytic cortex of TA muscles showed higher C/F after hypoxia (1.79 +/- 0.09 vs. 1.53 +/- 0.05 normoxia, P < 0.05), whereas the mainly oxidative TA core with smaller fibers showed no change in capillarity. The region of the SOL muscle with large-sized (mean fiber area 2,843 +/- 128 microm(2)) oxidative fibers (90% type I) had a higher C/F (by 30%) and CD (by 25%), whereas there was no angiogenesis in the region with sparse (76%) and smaller-sized (2,200 +/- 85 microm(2)) type I fibers. Thus systemic hypoxia differentially induces angiogenesis between and within hindlimb skeletal muscles, with fiber size contributing either directly (via a metabolic stimulus) or indirectly (via a mechanical stimulus) to the process. PMID- 11406492 TI - Patterns of wave break during ventricular fibrillation in isolated swine right ventricle. AB - Several different patterns of wave break have been described by mapping of the tissue surface during fibrillation. However, it is not clear whether these surface patterns are caused by multiple distinct mechanisms or by a single mechanism. To determine the mechanism by which wave breaks are generated during ventricular fibrillation, we conducted optical mapping studies and single cell transmembrane potential recording in six isolated swine right ventricles (RV). Among 763 episodes of wave break (0.75 times x s(-1) x cm(-2)), optical maps showed three patterns: 80% due to a wave front encountering the refractory wave back of another wave, 11.5% due to wave fronts passing perpendicular to each other, and 8.5% due to a new (target) wave arising just beyond the refractory tail of a previous wave. Computer simulations of scroll waves in three dimensional tissue showed that these surface patterns could be attributed to two fundamental mechanisms: head-tail interactions and filament break. We conclude that during sustained ventricular fibrillation in swine RV, surface patterns of wave break are produced by two fundamental mechanisms: head-tail interaction between waves and filament break. PMID- 11406493 TI - Regulation of tyrosine phosphorylation of PYK2 in vascular endothelial cells by lysophosphatidylcholine. AB - Lysophosphatidylcholine (LPC), a component of oxidized low-density lipoprotein, exerts various biological effects on vascular endothelial cells. However, the intracellular signaling of LPC is poorly understood. In this study, we investigated the involvement of proline-rich tyrosine kinase (PYK2) in LPC signaling in cultured bovine aortic endothelial cells by immunoprecipitation and Western blotting assays. Treatment of cells with LPC promoted a rapid increase in tyrosine phosphorylation of PYK2. LPC-stimulated PYK2 phosphorylation was inhibited by calcium chelators, 1,2-bis(2-aminophenoxy)ethane-N,N,N',N' tetraacetic acid-acetoxymethyl ester, EGTA, protein kinase C (PKC) inhibitor, GF 109203X, or PKC depletion by phorbol esters. PYK2 phosphorylation was inhibited by treatment with cytochalasin D but with neither botulinum C3 transferase nor overexpression of a dominant negative mutant of Rho A. LPC stimulated the association of Shc with PYK2, Shc tyrosine phosphorylation, and Grb2 binding to Shc and induced Ras activation. These results provide evidence that 1) LPC tyrosine phosphorylates PYK2 by calcium- and PKC-dependent mechanisms, 2) the intact cytoskeleton is required for LPC-stimulated PYK2 phosphorylation, and 3) LPC-activated Ras via the PYK2/Shc/Grb2 signaling. PMID- 11406494 TI - Bradykinin and des-Arg(9)-bradykinin metabolic pathways and kinetics of activation of human plasma. AB - In the serum of 116 healthy individuals, exogenous bradykinin (BK) half-life (27 +/- 10 s) was lower than that of des-Arg(9)-BK (643 +/- 436 s) and was statistically different in men compared with women. The potentiating effect of an angiotensin-converting enzyme (ACE) inhibitor was, however, more extensive for BK (9.0-fold) than for des-Arg(9)-BK (2.2- fold). The activities of ACE, aminopeptidase P (APP), and kininase I were respectively 44 +/- 12, 22 +/- 9, and 62 +/- 10 nmol x min(-1) x ml(-1). A mathematical model (y = kt(alpha)e(-beta t), t > 0), applied to the BK kinetically released from endogenous high-molecular weight kininogen (HK) during plasma activation in the presence of an ACE inhibitor, revealed a significant difference in the rate of formation of BK between men and women. For des-Arg(9)-BK, the active metabolite of BK, the rate of degradation was higher in women compared with men, correlating significantly with serum APP activity (r(2) = 0.6485, P < 0.001). In conclusion, these results constitute a basis for future pathophysiological studies of inflammatory processes where activation of the contact system of plasma and the kinins is involved. PMID- 11406495 TI - Age-associated changes in cardiovagal baroreflex sensitivity are related to central arterial compliance. AB - Cardiovagal baroreflex sensitivity (BRS) declines with advancing age in humans, but the underlying mechanism has not been established. Using two different approaches, we determined the relation between age-associated decline in cardiovagal BRS and the compliance of an artery in which arterial baroreceptors are located. First, we measured carotid artery compliance (via the simultaneous application of ultrasonography and arterial applanation tonometry) and cardiovagal BRS (phase IV of the Valsalva maneuver) in 47 healthy sedentary men that varied widely in age (19--76 yr). Cardiovagal BRS declined progressively with age (r = -0.69; P < or = 0.001) and was positively related to carotid artery compliance (r = 0.71; P < or = 0.001). Stepwise multiple-regression analysis revealed that carotid artery compliance was the strongest independent physiological correlate of cardiovagal BRS and that it explained 51% of the total variance. Second, we studied 13 middle-aged and older previously sedentary men (age 56 +/- 2 yr) before and after 13 wk of aerobic exercise intervention. Regular exercise increased both cardiovagal BRS and carotid artery compliance (P < 0.05) and the two events were strongly and positively related (r = 0.72; P < 0.01). We conclude that reduced carotid artery compliance may play an important mechanistic role in age-associated decrease in cardiovagal BRS in healthy sedentary humans. PMID- 11406496 TI - Protection of ischemic hearts by high glucose is mediated, in part, by GLUT-4. AB - Metabolic interventions that promote glucose use during ischemia have been shown to protect ischemic myocardium and improve functional recovery on reperfusion. We evaluated whether the cardioprotection afforded by high glucose during low-flow ischemia is associated with changes in the sarcolemmal content of glucose transporters, specifically GLUT-4. Isolated rat hearts were paced at 300 beats/min and perfused under normal glucose (5 mM) or high glucose (10 mM) conditions in buffer containing 0.4 mM albumin, 0.4 mM palmitate, and 70 mU/l insulin and subjected to 50 min of low-flow ischemia and 60 min of reperfusion. To determine the importance of insulin-sensitive glucose transporters in mediating cardioprotection, a separate group of hearts were perfused in the presence of cytochalasin B (10 microM), a preferential inhibitor of insulin sensitive glucose transporters. Ischemic contracture during low-flow ischemia and creatine kinase release on reperfusion was decreased, and the percent recovery of left ventricular function with reperfusion was enhanced in hearts perfused with high glucose (P < 0.03). Hearts perfused with high glucose exhibited increased GLUT-4 protein expression in the sarcolemmal membrane compared with control hearts under baseline conditions, and these changes were additive with low-flow ischemia. In addition, high glucose did not affect the baseline distribution of sarcolemmal GLUT-1 and blunted any changes with low-flow ischemia. These salutary effects were abolished when glucose transporters are blocked with cytochalasin B. These data demonstrate that protection of ischemic myocardium by high glucose is associated with increased sarcolemmal content of the insulin-sensitive GLUT-4 and suggest a target for the protection of jeopardized myocardium. PMID- 11406497 TI - Correlation of HO-1 expression with onset and reversal of hypoxia-induced vasoconstrictor hyporeactivity. AB - Rats exposed to chronic hypoxia (CH; 4 wk at 0.5 atm) exhibit attenuated renal vasoconstrictor reactivity to phenylephrine (PE). Preliminary studies from our laboratory suggest that this response is mediated by hypoxic induction of heme oxygenase (HO) and subsequent release of the endogenous vasodilator carbon monoxide. Because vascular HO mRNA is increased within hours of hypoxic exposure, we hypothesized that the onset of reduced reactivity may occur fairly rapidly and correlate with HO expression. Therefore, we examined the onset of attenuated vasoconstriction on CH exposure as well as the duration of hyporeactivity on return to a normoxic environment. Renal vascular resistance (RVR) responses to graded intravenous infusion of PE were measured in conscious rats under control conditions and after 24 h, 48 h, and 4 wk of CH exposure. Vasoreactivity responses were also determined in 4-wk CH rats 1, 5, 24, and 96 h after return to normoxia. We found that RVR responses to PE were significantly blunted after 48 h and 4 wk but not after 24 h of hypoxic exposure. Inhibition of HO with zinc protoporphyrin IX increased RVR and decreased renal blood flow in 48-h CH rats but not controls. Although reactivity to PE was gradually restored after 4 wk of CH, responsiveness was still slightly blunted at 96 h after return to normoxia. Western blot analysis demonstrated a correlation between HO-1 protein levels and attenuated vasoconstrictor response in CH and posthypoxic rats. These data suggest that the onset and offset of physiologically relevant vascular HO expression occur within 2--3 days. PMID- 11406498 TI - Restoration of vasodilation and CBF autoregulation by genistein in rat pial artery after brain injury. AB - This study determined whether, after fluid percussion injury (FPI), tyrosine kinase activation is coupled to inhibition of K(+) channels and alteration in cerebral blood flow (CBF) autoregulation in the rat pial artery. Injury of moderate severity (2--2.5 atm) was produced by FPI in anesthetized rats equipped with a closed cranial window. The suppressed vasodilation of the pial arterioles to calcitonin gene-related peptide (CGRP) and levcromakalim (LMK) and altered lower limit of CBF autoregulation after FPI were restored by genistein but not by daidzein, an inactive analog. Vasodilation to S-nitroso-N-acetyl penicillamine (0.1--10 micromol/l) was, however, little influenced after FPI. The restored vasodilation was decreased by sodium orthovanadate, suggesting the reciprocal action of tyrosine phosphorylation and dephosphorylation. After FPI, CGRP-induced vasodilation restored by genistein (10 micromol/l) was strongly antagonized by iberiotoxin but not by glibenclamide, whereas LMK-induced vasodilation was, in contrast, inhibited by glibenclamide but not by iberiotoxin. Taken together, we suggest that, after FPI, activation of tyrosine kinase links the inhibition of K(+) channels to impaired autoregulatory vasodilation in response to acute hypotension and alteration in CBF autoregulation in the rat pial artery. PMID- 11406499 TI - Nitric oxide generation by isolated descending vasa recta. AB - Nitric oxide (NO) generation by the outer medullary descending vasa recta (OMDVR) was measured with the fluorescent dye 4,5-diaminofluoroscein (DAF-2) during 30 min incubations. Addition of 0.1 or 1.0 mM L-arginine to the incubation buffer increased the DAF-2 signal by 8.7 and 13.6% (P = 0.08 and P < 0.05), respectively. Compared with L-arginine alone (0.1 mM), bradykinin (BK; 1 x 10(-7) M) enhanced the DAF-2 signal by 11.1% (P < 0.05). The NO synthase inhibitor N(omega)-nitro-L-arginine methyl ester (0.1 mM) reversed the BK-stimulated NO generation as measured with either DAF-2 or by the oxidation of Fe(2+) hemoglobin. Using 1 mM 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (tempol), a cell-permeant superoxide dismutase mimetic, we tested whether reduction of superoxide anion increases intracellular NO. Tempol increased DAF-2 fluorescence by 12 and 23.3%, respectively, over BK-stimulated or control vessels. Tempol also vasodilated ANG II (1 x 10(-8) M)-preconstricted OMDVR (P < 0.05). We conclude that NO generation by isolated OMDVR can be increased by L-arginine, that the endothelium-dependent vasodilator BK enhances NO production, and that NO consumption by superoxide plays a role in the determination of cellular NO concentrations. PMID- 11406500 TI - Glucocorticoid modulation of protein phosphorylation and sarcoplasmic reticulum function in rat myocardium. AB - To decipher the mechanism(s) underlying glucocorticoid action on cardiac contractile function, this study investigated the effects of adrenalectomy and dexamethasone treatment on the contents of sarcoplasmic reticulum (SR) Ca(2+) cycling proteins, their phosphorylation by endogenous Ca(2+)/calmodulin-dependent protein kinase II (CaM kinase II), and SR Ca(2+) sequestration in the rat myocardium. Cardiac SR vesicles from adrenalectomized rats displayed significantly diminished rates of ATP-energized Ca(2+) uptake in vitro compared with cardiac SR vesicles from control rats; in vivo administration of dexamethasone to adrenalectomized rats prevented the decline in SR function. Western immunoblotting analysis showed that the relative protein amounts of ryanodine receptor/Ca(2+)-release channel, Ca(2+)-ATPase, calsequestrin, and phospholamban were neither diminished significantly by adrenalectomy nor elevated by dexamethasone treatment. However, the relative amount of SR-associated CaM kinase II protein was increased 2.5- to 4-fold in dexamethasone-treated rats compared with control and adrenalectomized rats. Endogenous CaM kinase II activity, as judged from phosphorylation of ryanodine receptor, Ca(2+)-ATPase, and phospholamban protein, was also significantly higher (50--80% increase) in the dexamethasone-treated rats. The stimulatory effect of CaM kinase II activation on Ca(2+) uptake activity of SR was significantly depressed after adrenalectomy and greatly enhanced after dexamethasone treatment. These findings identify the SR as a major target for glucocorticoid actions in the heart and implicate modification of the SR CaM kinase II system as a component of the mechanisms by which dexamethasone influences SR Ca(2+)-cycling and myocardial contraction. PMID- 11406501 TI - P2 purinergic receptor activation enhances cardiac contractility in isolated rat and mouse hearts. AB - Activation of P2 purinergic receptors exerts a potent positive inotropic effect in the cardiac myocyte. However, it is unknown whether its activation can also cause an increased contractility in intact heart. With the use of isolated rat and mouse hearts, the objective of the present study was to investigate the effect of P2 receptor agonist on the function of the intact heart. In both Langendorff rat hearts and working rat and mouse heart models, the P2X receptor agonist 2-methylthio-ATP (2-meSATP) caused dose-dependent increases in left ventricular developed pressure, rate of contraction, and rate of relaxation. The extent of P2X receptor agonist-stimulated increase in contractility was significantly less than that stimulated by the beta-adrenergic agonist isoproterenol. However, the increase in contractility occurred without a significant effect on the basal heart rate, in contrast to that caused by isoproterenol. In isolated rat ventricular myocytes, both ATP and the P2X receptor agonist 2-meSATP stimulated large increases in the myocyte contractile amplitude (107 +/- 13% and 99 +/- 9%, n = 17 cells from 5 rats and n = 19 cells from 6 rats, respectively). 2-meSATP caused only a slight increase in phospholipase C activity and could stimulate myocyte contractility in the presence of phospholipase C inhibitor U-73122, consistent with the role of a phospholipase C-independent P2X receptor in mediating the positive inotropic effect of 2-meSATP. The data provide evidence for a potentially important physiological role of the cardiac P2X receptor and for the concept that agonist at this receptor may be beneficial for the treatment of cardiac dysfunction. PMID- 11406502 TI - Cysteinyl leukotrienes mediate enhanced vasoconstriction to angiotensin II but not endothelin-1 in SHR. AB - We assessed whether cysteinyl leukotrienes mediate the vasoconstrictor responses to angiotensin II and endothelin-1 in the mesenteric vascular bed of Wistar-Kyoto (WKY) and spontaneously hypertensive rats (SHR) perfused ex vivo at a constant flow rate of 5 ml/min with Krebs buffer. Maximal perfusion pressure response (E(max)) but not EC(50) values to angiotensin II (P < 0.001) and endothelin-1 (P < 0.01) were significantly higher in the SHR, whereas the responses to potassium chloride remained unchanged. Inclusion of the selective 5-lipoxygenase inhibitor AA-861 or the cysteinyl leukotriene receptor antagonist MK-571 significantly reduced the vasoconstrictor responses to angiotensin II but not to endothelin-1 and potassium chloride. The reduction in E(max) to angiotensin II was more pronounced in SHR (P < 0.001) than in WKY (P < 0.05) rats. Cysteinyl leukotrienes LTC(4)-, LTD(4)-, and LTE(4) (1 microM)-evoked vasoconstrictor responses were significantly higher in SHR (P < 0.05), whereas LTB(4) failed to evoke any response in either strain. These data suggest that 5-lipoxygenase metabolites, particularly cysteinyl leukotrienes, contribute to the exaggerated vasoconstrictor responses to angiotensin II but not to endothelin-1. PMID- 11406503 TI - Carbon monoxide produced by isolated arterioles attenuates pressure-induced vasoconstriction. AB - Studies were conducted on isolated rat gracilis muscle arterioles to examine the role of vascular heme oxygenase (HO)-derived carbon monoxide (CO) on myogenic constrictor responses to stepwise increments in intraluminal pressure. The arterioles express HO-2 but not HO-1 and manufacture CO. Both HO-2 protein expression and CO production are reduced in arterioles maintained for 18 h before experimentation in media containing HO-2 antisense oligodeoxynucleotides (AS ODN). Pressurization of arterioles mounted on a myograph over the pressure range of 40--100 mmHg elicits reduction of internal diameter. At pressures >40 mmHg, the internal diameter of vessels treated with either HO-2 AS-ODN, the HO inhibitor chromium mesoporphyrin (CrMP), or the K(+) channel blocker tetraethylammonium (TEA) are smaller than the corresponding control values. The inclusion of exogenous CO, but not of biliverdin, in the superfusion buffer attenuates pressure-induced vasoconstriction in CrMP-treated vessels. However, exogenous CO does not attenuate pressure-induced vasoconstriction in vessels treated with both CrMP and TEA. Collectively, these data suggest that CO of vascular origin attenuates pressure-induced arteriolar constriction via a mechanism involving a TEA-sensitive K(+) channel. PMID- 11406504 TI - Differential effects of protein kinase C on human vascular smooth muscle cell proliferation and migration. AB - Vascular smooth muscle cell (SMC) migration and proliferation contribute to intimal hyperplasia, and protein kinase C (PKC) may be required for both events. In this report, we investigated the role of PKC in proliferation and migration of SMC derived from the human saphenous vein. Activation of PKC by phorbol-12,13 dibutyrate (PDBu) or (-)-indolactam [(-)-ILV] increases SMC proliferation. Downregulation of PKC activity by prolonged incubation with phorbol ester or inhibition of PKC with chelerythrine in SMC diminished agonist-stimulated proliferation. In contrast, stimulation of PKC with PDBu or (-)-ILV inhibited basal and agonist-induced SMC chemotaxis. Moreover, downregulation of PKC or inhibition with chelerythrine accentuated migration. We postulated that the inhibitory effect of PKC on SMC chemotaxis was mediated through cAMP-dependent protein kinase (protein kinase A, PKA). In support of this hypothesis, we found that activation of PKC in SMC stimulated PKA activity. The cAMP agonist forskolin significantly inhibited SMC chemotaxis. Furthermore, the inhibitory effect of PKC on SMC chemotaxis was completely reversed by cAMP or PKA inhibitors. In search of the PKC isotype(s) underlying these differential effects of PKC in SMC, we identified eight isotypes expressed in human SMC. Only PKC-alpha, -beta I, delta, and -epsilon were eliminated by downregulation, suggesting that one or more of these four enzymes facilitate the observed phorbol ester-dependent effects of PKC in SMC. In summary, we found that PKC activation enhances proliferation but inhibits migration of human vascular SMC. These differential effect of PKC on vascular cells appears to be mediated through PKC-alpha, -beta I, -delta, and/or -epsilon. PMID- 11406505 TI - Contribution of perfusion pressure to vascular resistance response during head-up tilt. AB - We measured brachial and femoral artery flow velocity in eight subjects and peroneal and median muscle sympathetic nerve activity (MSNA) in five subjects during tilt testing to 40 degrees. Tilt caused similar increases in MSNA in the peroneal and median nerves. Tilt caused a fall in femoral artery flow velocity, whereas no changes in flow velocity were seen in the brachial artery. Moreover, with tilt, the increase in the vascular resistance employed (blood pressure/flow velocity) was greater and more sustained in the leg than in the arm. The ratio of the percent increase in vascular resistance in leg to arm was 2.5:1. We suggest that the greater vascular resistance effects in the leg were due to an interaction between sympathetic nerve activity and the myogenic response. PMID- 11406506 TI - Myocardial creatine kinase kinetics and isoform expression in hearts with severe LV hypertrophy. AB - Left ventricular (LV) hypertrophy (LVH) results in a fetal shift in myocardial creatine kinase (CK) expression. Because CK plays an important role in intracellular energy production, transport, and utilization, this study was performed to characterize changes in CK expression and CK flux in severe pressure overload LVH. Ascending aortic banding in 8-wk-old dogs resulted in LVH with a 92% increase in relative LV mass. In LVH hearts, CK-M isoform mRNA was decreased by 40% (P = 0.05) and protein was decreased by 50% (P < 0.01), whereas mitochondrial CK protein was decreased by 22% (P < 0.05). CK-B isoform mRNA was undetectable in normal hearts but was prominently expressed in LVH (P < 0.01); CK B protein was increased by more than 10-fold in LVH (P < 0.01). Despite these changes, total CK activity was normal in LVH. Myocardial CK flux was examined using (31)P magnetic resonance spectroscopy magnetization transfer. The CK forward rate constant was similar in normal and LVH hearts at baseline and did not change in either group during dobutamine treatment. In hearts with LVH, the CK forward flux rate was reduced by approximately 60% (P < 0.05) and decreased further during dobutamine. Thus, although pressure-overload LVH caused alterations of expression of both CK mRNA and protein levels, LV performance and oxygen consumption in response to dobutamine were normal. However, myocardial free ADP was increased in LVH hearts. This finding suggests that the CK alterations result in a need for higher ADP levels to maintain ATP synthesis in the hypertrophied heart. PMID- 11406507 TI - Frequency dependence of endocytosis in aortic baroreceptor neurons and role of group III mGluRs. AB - Synaptic transmission between baroreceptor afferents and the nucleus tractus solitarius (NTS) is known to exhibit frequency-dependent depression. Reductions in neurotransmitter release and alterations in mechanisms regulating synaptic transmission are hypothesized to be involved in the activity-dependent depression observed in baroreceptor afferent neurons. The present study utilized cultured aortic baroreceptor neurons and the fluorescent dyes FM1-43 and FM2-10 to characterize the process of endocytosis or vesicle retrieval and its dependence on 1) frequency of neuronal activation, 2) metabotropic glutamate receptor (mGluR) activation, and 3) calcium concentrations inside and outside the cell. Endocytosis per spike, measured in fluorescence units after a 10-s stimulus applied at frequencies of 0.5 (53 +/- 4), 1.0 (23 +/- 1), and 10.0 Hz (2.7 +/- 0.2), was significantly depressed at higher frequencies. Blockade of group III mGluRs with (RS)-cyclopropyl-4-phosphonophenylglycine (CPPG) facilitated endocytosis at all frequencies, suggesting that this receptor subtype may be involved in the inhibition of endocytosis. Manipulating the extracellular and intracellular calcium concentrations subsequent to exocytosis had no effect on endocytosis. These results suggest that frequency-dependent depression of endocytosis observed in vitro could contribute to the frequency-dependent depression of baroreceptor afferent neurotransmission and that group III mGluRs inhibit endocytosis. PMID- 11406508 TI - PTP-epsilon, a tyrosine phosphatase expressed in endothelium, negatively regulates endothelial cell proliferation. AB - The vascular endothelium is a dynamic interface between the blood vessel and circulating factors and, as such, plays a critical role in vascular events like inflammation, angiogenesis, and hemostasis. Whereas specific protein tyrosine kinases have been identified in these processes, less is known about their protein tyrosine phosphatase (PTP) counterparts. We utilized a RT PCR/differential hybridization assay to identify PTP-epsilon as a highly abundant endothelial cell PTP. PTP-epsilon mRNA expression is growth factor responsive, suggesting a role for this enzyme in endothelial cell proliferation. Overexpression of PTP-epsilon decreases proliferation by 60% in human umbilical vein endothelial cells (HUVEC) but not in smooth muscle cells or fibroblasts. In contrast, overexpression of PTP-epsilon (D284A), a catalytically inactive mutant, has no significant effect on HUVEC proliferation. These data provide the first functional characterization of PTP-epsilon in endothelial cells and identify a novel pathway that negatively regulates endothelial cell growth. Such a pathway may have important implications in vascular development and angiogenesis. PMID- 11406509 TI - Preconditioning attenuates apoptosis and necrosis: role of protein kinase C epsilon and -delta isoforms. AB - Preconditioning reduces cardiomyocyte necrosis in vivo and in vitro, but it is unknown whether preconditioning blocks apoptosis. We wanted to compare the effects of preconditioning on necrosis and apoptosis in cardiomyocytes. Necrosis was detected with propidium iodide, and apoptosis was quantified by three complementary techniques: flow cytometry, TdT-mediated dUTP nick-end labeling assay, and DNA-laddering electrophoresis. Apoptosis increased with simulated ischemia time (6 h, 19 +/- 1%; 12 h, 27 +/- 2%; 18 h, 40 +/- 4%; 24 h, 54 +/- 4%; and 36 h, 83 +/- 4%; n = 6 for each group). Simulated ischemia and reoxygenation contributed equally to apoptosis (12-h ischemia, 27 +/- 2%, n = 6; 12-h ischemia and 12-h reoxygenation, 51 +/- 4%, n = 6; and 24-h ischemia, 54 +/- 5%, n = 8). Necrosis occurred primarily during reoxygenation; none was detected during simulated ischemia. Preconditioning with 10 min of simulated ischemia reduced necrosis (18 +/- 6%, n = 8) but had no effect on apoptosis. However, three 1-min cycles of simulated ischemia separated by 5 min of reoxygenation reduced necrosis and apoptosis similarly. The protein kinase C (PKC) inhibitors Go6976 (0.1 microM) or chelerythrene (4 microM) abolished the effect of preconditioning. Preconditioning selectively activated PKC epsilon but had no effect on PKC delta and on total PKC enzyme activity. Preconditioning protected against necrosis and apoptosis, but the preconditioning ischemia required for blocking apoptosis was less than that for reducing necrosis. Activation of PKC epsilon isoform is important in mediating the protection. PMID- 11406510 TI - Characteristics of calcium sparks in cardiomyocytes derived from embryonic stem cells. AB - In embryonic stem (ES) cell-derived cardiomyocytes, spontaneous Ca(2+) sparks representing Ca(2+) release through ryanodine receptor (RyR) channels were characterized and correlated to the expression of RyRs as well as the Ca(2+) load of the sarcoplasmic reticulum (SR). In very early developmental stage (VEDS) cardiac precursor cells, global intracellular Ca(2+) concentration ([Ca(2+)](i)) fluctuations occurred, whereas Ca(2+) sparks and contractions were absent. In early developmental stages (EDS), contractions as well as Ca(2+) sparks were obvious. During the further differentiation to late developmental stage (LDS) cardiomyocytes, a marked increase in the frequency of global [Ca(2+)](i) transients, the amplitude and the frequency of Ca(2+) sparks, as well as the expression of RyRs and the volume of RyR-positive SR, was observed. Furthermore, the caffeine-releasable SR Ca(2+) load was elevated in LDS compared with EDS cardiomyocytes. A high-Ca(2+) solution raised spark frequency as well as amplitude in EDS cardiomyocytes to the levels of LDS cardiomyocytes. The characteristics of Ca(2+) sparks occurring in cardiomyocytes differentiated from ES cells may be governed by the Ca(2+) load of the SR and/or the density of RyRs. PMID- 11406511 TI - Ca(2+)-activated K(+) channels modulate basal and E(2)beta-induced rises in uterine blood flow in ovine pregnancy. AB - Uterine blood flow (UBF) increases >30-fold during ovine pregnancy. During the last trimester, this reflects vasodilation, which may be due to placentally derived estrogens. In nonpregnant ewes, estradiol-17 beta (E(2)beta) increases UBF >10-fold by activating nitric oxide synthase and large conductance calcium dependent potassium channels (BK(Ca)). To determine whether BK(Ca) channels modulate basal and E(2)beta-induced increases in UBF, studies were performed in near-term pregnant ewes with uterine artery flow probes and catheters for intra arterial infusions of tetraethylammonium (TEA), a selective BK(Ca) channel antagonist at <1 mM, in the absence or presence of E(2)beta (1 microg/kg iv). Uterine arteries were collected to measure BK(Ca) channel mRNA. TEA (0.15 mM) decreased basal UBF (P < 0.0001) 40 +/- 8% and 55 +/- 7% (n = 11) at 60 and 90 min, respectively, and increased resistance 175 +/- 48% without affecting (P > 0.1) mean arterial pressure (MAP), heart rate, or contralateral UBF. Systemic E(2)beta increased UBF 30 +/- 6% and heart rate 13 +/- 1% (P < or = 0.0001, n = 13) without altering MAP. Local TEA (0.15 mM) inhibited E(2)beta-induced increases in UBF without affecting increases in heart rate (10 +/- 4%; P = 0.006). BK(Ca) channel mRNA was present in uterine artery myocytes from pregnant and nonpregnant ewes. Exponential increases in ovine UBF in late pregnancy may reflect BK(Ca) channel activation, which may be mediated by placentally derived estrogens. PMID- 11406512 TI - Metabolism of S-nitrosoglutathione by endothelial cells. AB - S-nitrosoglutathione (GSNO) is an inhibitor of platelet aggregation and has also been shown to protect the ischemic heart from reperfusion-mediated injury. Although GSNO is often used in cell culture as a source of nitric oxide, the mechanisms of GSNO metabolism are not well established. We show here that GSNO decomposition by bovine aortic endothelial cells has an absolute dependence on the presence of cystine in the cell culture medium. In addition, GSNO decay is inhibited by diethyl maleate, an intracellular glutathione scavenger, but not by buthionine sulfoximine, a glutathione synthesis inhibitor. This indicates that thiols in general, rather than specifically glutathione, are the major factors that influence GSNO decay. Only 40% of the nitroso group of GSNO could be recovered as nitrite/nitrate, suggesting that the primary route of GSNO decay is reductive and that nitric oxide is only a minor product of GSNO decay. We conclude that the intracellular thiol pool causes the reduction of extracellular disulfides to thiols, which then directly reduce GSNO. PMID- 11406513 TI - Cardiac myocytes exposed to anoxia-reoxygenation promote neutrophil transendothelial migration. AB - The goal of the present study was to assess whether cardiac myocytes exposed to anoxia-reoxygenation (A/R) could generate a chemotactic gradient for polymorphonuclear neutrophil (PMN) transendothelial migration. Exposure of neonatal mouse cardiac myocytes to A/R induced an oxidant stress in the myocytes. Supernatants obtained from A/R-conditioned myocytes promoted mouse PMN migration across mouse myocardial endothelial cell monolayers. This increase in PMN transendothelial migration could be prevented if catalase or a platelet activating factor (PAF) antagonist was added to the supernatants before assay. Supernatants from A/R-conditioned myocytes activated endothelial cells by inducing an intracellular oxidant stress. The oxidant stress and PMN transendothelial migration induced by supernatants from A/R-conditioned myocytes were substantially reduced when endothelial cells derived from manganese superoxide dismutase overexpressing mice were used in the assays. Supernatants from A/R-conditioned myocytes also increased endothelial cell surface levels of E selectin and intercellular adhesion molecule-1. Our results indicate that cardiac myocytes exposed to A/R can generate a chemotactic gradient, presumably due to production and release of stable oxidants and PAF. The ability of supernatants from A/R-conditioned myocytes to promote PMN transendothelial migration was largely dependent on induction of an oxidant stress in endothelial cells. In addition, these supernatants also induced a proadhesive phenotype in the endothelial cells. PMID- 11406514 TI - O(2) release from erythrocytes flowing in a narrow O(2)-permeable tube: effects of erythrocyte aggregation. AB - The effects of erythrocyte aggregation on O(2) release were examined using O(2) permeable fluorinated ethylenepropylene copolymer tubes (inner diameter, 25 microm; outer diameter, 100 microm). Measurements were performed using an apparatus built on an inverted microscope that contained a scanning-grating spectrophotometer with a photon count detector connected to two photomultipliers and an image processor through a video camera. The rate of O(2) release from the cells flowing in the narrow tube was determined based on the visible absorption spectrum and the flow velocity of the cells as well as the tube size. When the tube was exposed to nitrogen-saturated deoxygenated saline containing 10 mM sodium dithionite, the flowing erythrocytes were deoxygenated in proportion to the traveling distance, and the deoxygenation at a given distance increased with decreasing flow velocity and cell concentration (hematocrit). Adding Dextran T-70 to the cell suspension increased erythrocyte aggregation in the tube, which resulted in suppressed cell deoxygenation and increased marginal cell-free-layer thickness. The deoxygenation was inversely proportional to the cell-free-layer thickness. The relation was not essentially altered even when the medium viscosity was adjusted with Dextran T-40 to remain constant. The rate of O(2) release from erythrocytes in the tube was discussed in relation to the O(2) diffusion process. We conclude that the diffusion of O(2) from erythrocytes flowing in narrow tubes is inhibited primarily by erythrocyte aggregation itself and partly by thickening of the cell-free layer. PMID- 11406515 TI - Expression of metabotropic glutamate receptors in nodose ganglia and the nucleus of the solitary tract. AB - The purpose of this study was to identify the complement of metabotropic glutamate receptors (mGluRs) expressed in nodose ganglia and the nucleus tractus solitarius (NTS). mRNA from these tissues was isolated and amplified with standard RT-PCR with primers specific for each mGluR subtype. The results of this analysis showed that the NTS expresses all eight mGluR subtypes, whereas nodose ganglia express only group III mGluRs: mGluR4, mGluR6, mGluR7, and mGluR8. Application of the group III-specific mGluR agonist L-(+)-2-amino-4 phosphonobutyric acid (100 microM) reversibly inhibited voltage-gated calcium currents isolated from DiI-labeled aortic baroreceptor neurons and unlabeled nodose neurons. The results of this study suggest that group III mGluRs are the primary mGluR subtype expressed in visceral afferent neurons and that these receptors may be involved in afferent central transmission. PMID- 11406516 TI - Guidelines for musculoskeletal ultrasound in rheumatology. PMID- 11406517 TI - Debilitating knee pain in a patient with "normal" radiographs. PMID- 11406518 TI - Breast implants and illness: a model of psychological factors. AB - Studies of disease outcomes have not produced an explanation or an intervention for the symptoms and complaints that some women have attributed to breast implants. Reviews of the literature have found no increased risk of specific systemic disease, and no treatment recommendations have emerged. However, similar symptoms in fibromyalgia, chronic fatigue, and other contexts have been considered to be stress or behaviourally mediated, and a number of promising behavioural interventions have been developed. Aetiological, research, and treatment implications may follow from the consideration of such symptoms within a behavioural medicine model that allows for the interaction of physical and psychological influences. In the case of implants, a mass somatisation model may also help to discern the potential effects of litigation and other social influences. PMID- 11406520 TI - Effects of treatment with a fully human anti-tumour necrosis factor alpha monoclonal antibody on the local and systemic homeostasis of interleukin 1 and TNFalpha in patients with rheumatoid arthritis. AB - OBJECTIVES: To study the short term effects of a single dose of D2E7, a fully human anti-tumour necrosis factor (TNFalpha) monoclonal antibody (mAb), on the local and systemic homeostasis of interleukin 1beta (IL1beta) and TNFalpha in patients with rheumatoid arthritis (RA). METHODS: All patients with RA enrolled in a phase I, single dose, placebo controlled study with D2E7 in our centre were studied. Systemic cytokine levels, acute phase reactants, and leucocyte counts were studied at days 0, 1, and 14 after the first administration of anti-TNF mAb (n=39) or placebo (n=11). The cellularity and the expression of IL1 and TNFalpha in synovial tissue were studied in knee biopsy specimens obtained at baseline and at day 14 in 25 consenting patients. RESULTS: A single dose of anti-TNF mAb induced a rapid clinical improvement, a decrease in acute phase reaction, and increased lymphocyte counts in patients with active RA. The protein levels of IL1beta in the circulation were low and remained unchanged, but the systemic levels of IL1beta mRNA (p=0.002) and the concentrations of IL1 receptor antagonist (IL1ra) and IL6 (p=0.0001) had already dropped within 24 hours and this persisted up to day 14. Systemic levels of TNFalpha mRNA were low and remained unchanged, though total TNFalpha (free and bound) in the circulation increased after D2E7, probably reflecting the presence of TNF-antiTNF mAb complexes (p<0.005, at days 1 and 14). Both TNF receptors dropped below baseline levels at day 14 (p<0.005). Despite clinical improvement of arthritis, no consistent immunohistological changes were seen two weeks after anti-TNF administration. Endothelial staining for IL1beta tended to decrease in treated patients (p=0.06) but not in responders. The staining for IL1beta and TNFalpha in sublining layers and vessels was mutually correlated (r(s)=0.47 and 0.58 respectively, p<0.0005) and the microscopic scores for inflammation correlated with sublining TNFalpha and IL1beta scores (r(s)=0.65 and 0.54 respectively, p<0.0001), though none of these showed significant changes during the study. CONCLUSIONS: Blocking TNFalpha in RA results in down regulation of IL1beta mRNA at the systemic level and in reduction of the endogenous antagonists for IL1 and TNF and of other cytokines related to the acute phase response, such as IL6, within days. At the synovial level, anti-TNF treatment does not modulate IL1beta and TNFalpha in the short term. The synovial expression of these cytokines does not reflect clinical response to TNF neutralisation. PMID- 11406521 TI - Assessment of phalangeal bone loss in patients with rheumatoid arthritis by quantitative ultrasound. AB - OBJECTIVE: Periarticular osteopenia is an early radiological sign of rheumatoid arthritis (RA). Quantitative ultrasound (QUS) devices have recently been shown to be useful for assessing osteoporosis. In this study the capability of a transportable and easy to use QUS device to detect skeletal impairment of the finger phalanges in patients with RA was investigated. METHODS: In a cross sectional study 83 women (30 controls, 29 with glucocorticosteroid (GC) treated RA, and 24 with GC treated vasculitis) were examined. QUS measurements were obtained at the metaphyses of the proximal phalanges II-V and directly at the proximal interphalangeal joints II-IV with a DBM Sonic 1200 (IGEA, Italy) QUS device. Amplitude dependent speed of sound (AD-SoS) was evaluated. In 23 of the patients with RA, hand radiographs were evaluated. RESULTS: Significant differences between patients with RA and the other groups were found for AD-SoS at both measurement sites. Compared with age matched controls, the AD-SoS of patients with RA was lowered by two and three standard deviations at the metaphysis and joint, respectively. Fingers of patients with RA without erosions (Larsen score 0-I) already had significantly decreased QUS values, which deteriorated further with the development of erosions (Larsen II-V). CONCLUSION: This study indicates that QUS is sensitive to phalangeal periarticular bone loss in RA. QUS is a quick, simple, and inexpensive method free of ionising radiation that appears to be suited to detection of early stages of periarticular bone loss. Its clinical use in the assessment of early RA should be further evaluated in prospective studies. PMID- 11406522 TI - Importance of NF-kappaB in rheumatoid synovial tissues: in situ NF-kappaB expression and in vitro study using cultured synovial cells. AB - OBJECTIVES: To examine whether inhibition of NF-kappaB induces apoptosis of human synovial cells stimulated by tumour necrosis factor alpha (TNFalpha), interleukin 1beta (IL1beta), and anti-Fas monoclonal antibody (mAb). METHODS: The expression of proliferating cell nuclear antigen (PCNA), NF-kappaB, and the presence of apoptotic synovial cells were determined in synovial tissues. Apoptosis of cultured synovial cells was induced by inhibition of NF-kappaB nuclear translocation by Z-Leu-Leu-Leu-aldehyde (LLL-CHO). The activation of caspase-3 and expression of XIAP and cIAP2 in synovial cells in LLL-CHO induced apoptosis was also examined. RESULTS: Abundant PCNA+ synovial cells were found in rheumatoid arthritis (RA) synovial tissue, though a few apoptotic synovial cells were also detected in the RA synovial tissues. Nuclear NF-kappaB was expressed in RA synovial cells. Electrophoretic mobility shift assay showed that treatment of cells with TNFalpha or IL1beta significantly stimulated nuclear NF-kappaB activity. A small number of apoptotic synovial cells expressing intracellular active caspase-3 were found after treatment of cells with LLL-CHO. Although treatment of RA synovial cells with TNFalpha or IL1beta alone did not induce apoptosis, apoptosis induced by LLL-CHO and caspase-3 activation were clearly enhanced in TNFalpha or IL1beta stimulated synovial cells compared with unstimulated synovial cells. Furthermore, induction of apoptosis of synovial cells with caspase-3 activation by anti-Fas mAb was clearly increased by LLL-CHO. The expression of cIAP2 and XIAP in synovial cells may not directly influence the sensitivity of synovial cells to apoptosis induced by LLL-CHO. CONCLUSION: The results suggest that NF-kappaB inhibition may be a potentially important therapeutic approach for RA by correcting the imbalance between apoptosis and proliferation of synovial cells in RA synovial tissue. PMID- 11406523 TI - CpG-DNA derived from sera in systemic lupus erythematosus enhances ICAM-1 expression on endothelial cells. AB - OBJECTIVE: To examine the effect of transfection of oligodeoxynucleotides (ODNs) containing a CpG motif (CpG-ODN), of which the sequence was derived from circulating DNA in the sera of patients with systemic lupus erythematosus (SLE), on the expression of intercellular adhesion molecule-1 (ICAM-1) and synthesis of mRNA for proinflammatory cytokines and ICAM-1 in human umbilical vein endothelial cells (EC). METHODS: A CpG-ODN or a control analogue, GpC-ODN, was transfected into EC. ICAM-1 expression was examined by flow cytometry, and expression of mRNA in EC encoding interleukin 1 (IL1), IL6, IL8, tumour necrosis factor alpha (TNFalpha), interferon gamma (IFNgamma), and ICAM-1 was examined by semiquantitative reverse transcriptase-polymerase chain reaction. RESULTS: The CpG-ODN augmented the expression of ICAM-1 on EC determined by flow cytometry and increased mRNA levels of IL6, IL8, TNFalpha, IFNgamma, and ICAM-1, but the GpC ODN did not. CONCLUSION: Synthesised DNA, with a sequence corresponding to that of the fragment containing the CpG motif, in sera of patients with SLE was found to enhance ICAM-1 expression on EC, suggesting the participation of circulating DNA fragments in the pathogenesis of vasculitis in SLE. PMID- 11406524 TI - Quantitative ultrasonography in rheumatoid arthritis: evaluation of inflammation by Doppler technique. AB - OBJECTIVE: To evaluate ultrasonographic methods, including the Doppler technique, as measures of synovial inflammation in finger joints of patients with rheumatoid arthritis. METHODS: Ultrasonography was performed with a high frequency transducer (13 MHz). Evaluation of the sonographic data was conducted by two independent observers and included measurement of synovial area and thickness (grey tone ultrasound), vascularisation (power/colour Doppler), and indices of the intra- and extrasynovial arterial flow (spectral Doppler). The flow pattern was estimated by the indices of pulsatility (PI) and resistance (RI). RESULTS: The sonographic measurements of joint space were reproducible with intraobserver, intraclass correlation coefficients (ICC) 0.82-0.97 (p<0.0001) and interobserver ICC 0.81 (p<0.0001). The mean (SD) fraction of the synovium vascularised in the patients was 0.15 (0.15). The synovial blood flow was characterised by a diastolic flow-that is, the flow persisting during the diastole. The mean (SD) PI was 1.92 (1.18) and RI 0.70 (0.13). The estimated vascular fraction correlated with the erythrocyte sedimentation rate (ESR) (r(s)=0.53, p=0.03). The relative Pi (rPi), an estimate of an abnormally low resistance to vascularisation, correlated with both ESR (r(s)=-0.557, p<0.05) and Health Assessment Questionnaire score (r(s)=-0.584, p<0.05). After an injection of contrast Levovist the vascular fraction increased, while no difference in PI and RI was observed. CONCLUSION: Ultrasonography is a reliable tool for estimating the size of the joint space and the synovial activity measured by the degree of vascularisation and pattern of flow. Ultrasonography may be useful in monitoring the synovial inflammation in rheumatoid arthritis. PMID- 11406525 TI - Concomitant diagnosis of primary Sjogren's syndrome and systemic AL amyloidosis. AB - A 48 year old woman was referred to hospital for buccal discomfort. Physical examination showed a macroglossia and features of xerostomia. She was diagnosed as having primary Sjogren's syndrome according to the criteria proposed by the European Community study group in 1993. Furthermore, a lower lip salivary gland biopsy showed amyloid deposits that were also seen in the stomach and in the bone marrow. Echocardiography was consistent with cardiac amyloidosis. Serum immunofixation identified a monoclonal IgGlambda. As far as is known, this is the first report of systemic primary amyloidosis associated with Sjogren's syndrome. The relation between these two disorders is discussed. PMID- 11406526 TI - Human immunodeficiency virus associated spondyloarthropathy: pathogenic insights based on imaging findings and response to highly active antiretroviral treatment. AB - The pathogenesis of human immunodeficiency virus (HIV) associated spondyloarthropathy (SpA) is poorly understood. In this case report a patient is described with severe HIV associated reactive arthritis, who on magnetic resonance imaging and sonographic imaging of inflamed knees had extensive polyenthesitis and adjacent osteitis. The arthritis deteriorated despite conventional antirheumatic treatment, but improved dramatically after highly active antiretroviral treatment, which was accompanied by a significant rise in CD4 T lymphocyte counts. The implications of the localisation of pathology and effect of treatment for pathogenic models of SpA and rheumatoid arthritis in the setting of HIV infection are discussed. PMID- 11406527 TI - Severe atherosclerotic changes, including aortic occlusion, associated with hyperhomocysteinaemia and antiphospholipid antibodies. AB - Three patients are described with severe systemic atherosclerosis, including aortic occlusion, in the presence of a spectrum of risk factors, including hypercholesterolaemia, hypertension, a positive family history of cardiovascular problems, and hyperhomocysteinaemia. In all three patients high levels of anticardiolipin antibodies were found. The possible pathogenic role of antiphospholipid antibodies in atherosclerosis in the context of hyperhomocysteinaemia in these patients is discussed. PMID- 11406528 TI - Haemopoietic stem cell transplantation in the treatment of severe autoimmune diseases 2000. AB - An international meeting took place in Basel, Switzerland from 5 to 7 October 2000 involving 180 participants from 30 countries, with the aim of assessing the existing data on autologous haemopoietic stem cell transplantation (HSCT) in the treatment of severe autoimmune disease, and to decide on future trial planning. Data on 390 patients were presented: 260 from the EBMT/EULAR Basel European/Asian database, 87 from North America (55 from the IBMTR), 39 from Australia, and 4 others. The major disease categories and number of patients receiving transplant were: multiple sclerosis (MS) 127, systemic sclerosis (SSc) 72, rheumatoid arthritis (RA) 70, juvenile idiopathic arthritis (JIA) 36, systemic lupus erythematosus (SLE) 34, dermatomyositis/polymyositis (DM/PM) 5, idiopathic thrombocytopenic purpura (ITP) 7. Single or several cases of other autoimmune diseases were reported. Clinically significant responses were seen in two thirds of all the cases and in all disease categories, with a more accentuated trend towards relapse in JIA and RA. Treatment was associated with a significant morbidity and mortality. In the EULAR/EBMT database (71 centres in 22 countries), a mobilisation associated mortality of 1.5% and an overall procedure related mortality (actuarially adjusted at 12 months) of 9% (confidence interval 6 to 12%) were found, with significant variation between diseases. The North American data showed similar results. Higher mortalities were seen in SSc and systemic JIA, with only one death reported in RA. After presentation of the data and workshop discussion a consensus was reached on several aspects: prospective randomised phase III trials are now appropriate in SSc, MS, and RA. A protocol is ready for SSc (ASTIS Trial), concepts are clear for MS and RA. Further phase I and II data are required in SLE, JIA, and vasculitis. The need for continuing collection of all cases after mobilisation by the standardised EBMT and IBMTR data forms was emphasised. PMID- 11406530 TI - Who is the boss in the retinoblastoma family? The point of view of Rb2/p130, the little brother. AB - This review portrays an updated overview about the possible tumor suppressive properties of the Rb2/p130 gene, the third member of the retinoblastoma (RB) family of genes, including RB itself and p107. After a brief analysis of the established structural and functional similarities among the three genes, the main purpose is to critically analyze present evidence whether Rb2/p130 shares the role of a tumor suppressor. Taking into account the well-proven growth suppressive properties of Rb2/p130 and p107, we discuss the analysis of mutated or deleted forms of Rb2/p130 found in a number of human cancers. Finally, we take into consideration the data provided by the targeted disruption of each RB family gene, alone or in combination, in the mouse model. PMID- 11406531 TI - Heparanase expression in primary and metastatic pancreatic cancer. AB - The human endoglycosidase heparanase (hpa) degrades heparan-sulfate proteoglycans, which constitute prominent components of basement membranes and extracellular matrix. Due to the critical function of hpa in cancer cell invasion and metastasis, we have analyzed the expression of hpa in human primary and metastatic pancreatic cancer as well as in the normal pancreas and in chronic pancreatic inflammation. By real-time quantitative PCR, there was a 7.9- and 30.2 fold increase of hpa mRNA in chronic pancreatitis and pancreatic cancer tissue samples, respectively, in comparison with normal pancreatic tissues. There was a significant correlation between enhanced hpa mRNA expression and shorter postoperative patient survival. hpa mRNA and protein localized in the cancer cells of primary and metastatic pancreatic cancer, with a preferentially higher expression at the primary tumor site. Cultured pancreatic cancer cells transfected with a full-length hpa construct displayed enhanced invasiveness in an invasion chamber assay. These results suggest that hpa overexpression in human pancreatic cancers facilitates cancer cell invasion, thereby enhancing the metastatic potential of the tumors. PMID- 11406532 TI - Prostate stem cell antigen is overexpressed in human transitional cell carcinoma. AB - Prostate stem cell antigen (PSCA), a homologue of the Ly-6/Thy-1 family of cell surface antigens, is expressed by a majority of human prostate cancers and is a promising target for prostate cancer immunotherapy. In addition to its expression in normal and malignant prostate, we recently reported that PSCA is expressed at low levels in the transitional epithelium of normal bladder. In the present study, we compared the expression of PSCA in normal and malignant urothelial tissues to assess its potential as an immunotherapeutic target in transitional cell carcinoma (TCC). Immunohistochemical analysis of PSCA protein expression was performed on tissue sections from 32 normal bladder specimens, as well as 11 cases of low-grade transitional cell dysplasia, 21 cases of carcinoma in situ (CIS), 38 superficial transitional cell tumors (STCC, stages T(a)-T(1)), 65 muscle-invasive TCCs (ITCCs, stages T(2)-T(4)), and 7 bladder cancer metastases. The level of PSCA protein expression was scored semiquantitatively by assessing both the intensity and frequency (i.e., percentage of positive tumor cells) of staining. We also examined PSCA mRNA expression in a representative sample of normal and malignant human transitional cell tissues. In normal bladder, PSCA immunostaining was weak and confined almost exclusively to the superficial umbrella cell layer. Staining in CIS and STCC was more intense and uniform than that seen in normal bladder epithelium (P < 0.001), with staining detected in 21 (100%) of 21 cases of CIS and 37 (97%) of 38 superficial tumors. PSCA protein was also detected in 42 (65%) of 65 of muscle-invasive and 4 (57%) of 7 metastatic cancers, with the highest levels of PSCA expression (i.e., moderate-strong staining in >50% of tumor cells) seen in 32% of invasive and 43% of metastatic samples. Higher levels of PSCA expression correlated with increasing tumor grade for both STCCs and ITCCs (P < 0.001). Northern blot analysis confirmed the immunohistochemical data, showing a dramatic increase in PSCA mRNA expression in two of five muscle-invasive transitional cell tumors when compared with normal samples. Confocal microscopy demonstrated that PSCA expression in TCC is confined to the cell surface. These data demonstrate that PSCA is overexpressed in a majority of human TCCs, particularly CIS and superficial tumors, and may be a useful target for bladder cancer diagnosis and therapy. PMID- 11406533 TI - The paired box domain gene PAX5 is fused to ETV6/TEL in an acute lymphoblastic leukemia case. AB - The PAX5 gene, encoding the B-cell-specific activator protein, is a critical determinant of commitment to the B-lymphocyte pathway. This gene, mapped at 9p13, is juxtaposed to the immunoglobulin heavy chain (IgH) gene as a result of the t(9;14)(p13;q32), a rare but recurring translocation found in a subset of B-cell non-Hodgkin's lymphoma cases. In all of these, this translocation results in deregulated expression of the gene product because of the proximity of IgH. We present here the molecular characterization of a previously reported acute lymphoblastic leukemia case carrying a t(9;12)(q11;p13) translocation. Using 5' rapid amplification of cDNA ends PCR, a novel chimeric transcript was identified that contained the NH(2)-terminal region of PAX5 and most of the ETV6/TEL gene on 12p13. According to the fusion transcript, the resulting chimeric protein would retain the PAX5 paired-box domain and both the helix-loop-helix and DNA binding domains of TEL. Thus, it is reasonable to hypothesize that this protein could act as an aberrant transcription factor. This is the first report of PAX5 rearrangement in a human malignancy resulting in a chimeric transcript. PMID- 11406534 TI - Ny-ESO-1 expression and immunogenicity associated with transitional cell carcinoma: correlation with tumor grade. AB - NY-ESO-1 mRNA expression in transitional cell carcinoma was investigated by reverse transcription-PCR and immunohistochemistry. NY-ESO-1 mRNA was detected in 20 of 62 (32%) tumor specimens. There was a correlation between NY-ESO-1 expression and tumor grade: 0 of 4 (0%) grade 1 (G1), 6 of 26 (23%) grade 2 (G2), and 14 of 32 (44%) grade 3 (G3) tumors were NY-ESO-1 mRNA positive. Immunohistochemical analysis using NY-ESO-1-specific monoclonal antibody ES121 showed that 2 of 14 NY-ESO-1 mRNA-expressing G3 tumors were positive for NY-ESO 1. No NY-ESO-1 staining was observed in the panel of 30 G1 or G2 tumor specimens, including 6 NY-ESO-1 mRNA-positive cases. Sera from an expanded panel of 124 patients with transitional cell carcinoma were tested for the presence of NY-ESO 1 antibody. Seropositivity was observed in 9 of 72 (12.5%) patients with G3 tumors, whereas none of 52 patients with G1 or G2 tumors produced antibody against NY-ESO-1. In the 9 positive patients with NY-ESO-1 antibody, 4 had muscular invasive tumors, and 5 had carcinoma in situ. PMID- 11406535 TI - Analysis of circulating tumor DNA in plasma at diagnosis and during follow-up of lung cancer patients. AB - We evaluated whether the amount of circulating DNA in plasma could discriminate between lung cancer patients and healthy individuals and whether it is related to disease progression, and we analyzed the kinetics of plasma DNA in disease-free, surgically resected patients. Plasma DNA quantification and analysis of microsatellite alterations were performed in a consecutive series of 84 patients with non-small cell lung cancer, who were studied during follow-up, and 43 healthy controls. In patients, the mean values of plasma DNA concentration were higher than in controls even considering stage Ia patients. Sensitivity and specificity estimates were calculated as the area under the receiver operating characteristic curve (AUC-ROC) curve and showed a value of 0.844. Variations in DNA level and in microsatellite changes correlated with the clinical status of 38 patients monitored during follow-up. The data suggest that quantification and molecular characterization of plasma DNA in lung cancer patients are valuable noninvasive diagnostic tools for discriminating patients from unaffected individuals and for detecting early recurrence during follow-up. PMID- 11406536 TI - Transforming growth factor-beta receptor type I gene is frequently mutated in ovarian carcinomas. AB - Ovarian carcinomas (OCs), particularly recurrent OCs, are frequently resistant to transforming growth factor (TGF)-beta-mediated growth inhibition. Mutations in the TGF-beta receptor type II (TbetaR-II) gene are only evident in a minority of OCs, suggesting that other alterations of the TGF-beta signaling pathway may be involved in OC. Using PCR, cold single-strand conformation polymorphism, and DNA sequencing, we now show that 33% of primary OCs (10 of 30) harbor somatic changes in exons 2, 3, 4, and 6 of the TGF-beta receptor I (TbetaR-I) gene. Most of the changes are missense mutations and clustered largely in the catalytic domain of the receptor kinase. Interestingly, seven additional cases (23.3%) showed heterozygous carriers of an allelic variant [a 9-nucleotide deletion, del(GGC)(3)] in exon 1 of the TbetaR-I gene. This is in contrast with 10.6% of del(GGC)(3) heterozygous carriers in a recent report of a large normal population (n = 735; B. Pasche et al., Cancer Res., 59: 5678-5682, 1999). These results indicate that TbetaR-I is frequently mutated in OC and suggest that resistance to TGF-beta-mediated growth inhibition may frequently involve alterations of the TbetaR-I gene. PMID- 11406537 TI - Human prostate cancer and benign prostatic hyperplasia: molecular dissection by gene expression profiling. AB - Critical aspects of the biology and molecular basis for prostate malignancy remain poorly understood. To reveal fundamental differences between benign and malignant growth of prostate cells, we performed gene expression profiling of primary human prostate cancer and benign prostatic hyperplasia (BPH) using cDNA microarrays consisting of 6500 human genes. Frozen prostate specimens were processed to facilitate extraction of RNA from regions of tissue enriched in either benign or malignant epithelial cell growth within a given specimen. Gene expression in each of the 16 prostate cancer and nine BPH specimens was compared with a common reference to generate normalized measures for each gene across all of the samples. Using an analysis of complete pairwise comparisons of expression profiles among all of the samples, we observed clearly discernable patterns of overall gene expression that differentiated prostate cancer from BPH. Further analysis of the data identified 210 genes with statistically significant differences in expression between prostate cancer and BPH. These genes include many not recognized previously as differentially expressed in prostate cancer and BPH, including hepsin, which codes for a transmembrane serine protease. This study reveals for the first time that significant and widespread differences in gene expression patterns exist between benign and malignant growth of the prostate gland. Gene expression analysis of prostate tissues should help to disclose the molecular mechanisms underlying prostate malignant growth and identify molecular markers for diagnostic, prognostic, and therapeutic use. PMID- 11406538 TI - Promoter hypermethylation of the DNA repair gene O(6)-methylguanine-DNA methyltransferase is associated with the presence of G:C to A:T transition mutations in p53 in human colorectal tumorigenesis. AB - Defects in DNA repair may be responsible for the genesis of mutations in key genes in cancer cells. The tumor suppressor gene p53 is commonly mutated in human cancer by missense point mutations, most of them G:C to A:T transitions. A recognized cause for this type of change is spontaneous deamination of the methylcytosine. However, the persistence of a premutagenic O(6)-methylguanine can also be invoked. This last lesion is removed in the normal cell by the DNA repair enzyme O(6)-methylguanine-DNA methyltransferase (MGMT). In many tumor types, epigenetic silencing of MGMT by promoter hypermethylation has been demonstrated and linked to the appearance of G to A mutations in the K-ras oncogene in colorectal tumors. To study the relevance of defective MGMT function by aberrant methylation in relation to the presence of p53 mutations, we studied 314 colorectal tumors for MGMT promoter hypermethylation and p53 mutational spectrum. Inactivation of MGMT by aberrant methylation was associated with the appearance of G:C to A:T transition mutations at p53 (Fischer's exact test, two-tailed; P = 0.01). Overall, MGMT methylated tumors displayed p53 transition mutations in 43 of 126 (34%) cases, whereas MGMT unmethylated tumors only showed G:C to A:T changes in 37 of 188 (19%) tumors. A more striking association was found in G:C to A:T transitions in non-CpG dinucleotides; 71% (12 of 17) of the total non-CpG transition mutations in p53 were observed in MGMT aberrantly methylated tumors (Fischer's exact test, two-tailed; P = 0.008). Our data suggest that epigenetic silencing of MGMT by promoter hypermethylation may lead to G:C to A:T transition mutations in p53. PMID- 11406539 TI - Prostate carcinomas developing in transgenic rats with SV40 T antigen expression under probasin promoter control are strictly androgen dependent. AB - We have generated a transgenic rat with the SV40 T antigen under probasin promoter control, allowing prostate-specific gene expression. Males demonstrate atypical epithelial cell proliferation in the prostate from 4 weeks of age and develop prostate carcinomas at 100% incidence before they are 15 weeks old. Castration at 5 weeks of age was found to inhibit the prostate tumor formation completely, whereas testosterone propionate administration induced marked cell proliferation as well as microinvasion in prostate carcinomas. Castration at 20 weeks of age, after tumor development, even with testosterone propionate treatment, induced complete tumor involution within 5 weeks. To investigate the underling processes, sequential histological changes were monitored 1, 2, 3, 7, 14, and 21 days after castration. At days 1-3, many apoptotic bodies and inflammatory cells, including foam cells, were observed, and clear glandular structures were no longer evident in the tumors. Seven days after castration, most glands were involved, and nuclei of the cells did not show atypia. After 14 and 21 days, only atrophic glands were observed. During this process, expression of caspase 3, caspase 6, BAX, bcl-x, TRPM-2, and MMP7 genes was apparently increased. Comparison of the gene expression profile between a prostate carcinoma in a transgenic animal and a normal prostate of a wild-type rat by a cDNA array technique was also conducted. The results suggested that our model is suitable to investigate mechanisms of carcinogenesis, including androgen dependence, involution, and apoptosis. PMID- 11406540 TI - Potentiation of nitric oxide-induced apoptosis of MDA-MB-468 cells by farnesyltransferase inhibitor: implications in breast cancer. AB - High amounts of nitric oxide (NO) produced by activated macrophages or NO donors are required to induce cytotoxicity and apoptosis in pathogens and tumor cells. High concentrations of NO may lead to nonspecific toxicity thereby limiting the use of NO donors in the treatment of cancer. In this study, we tested the possibility of potentiating the apoptotic action of NO in a human breast cancer cell line, MDA-MB-468, by combining it with a farnesyltransferase inhibitor (FTI), which has been shown to induce apoptosis in some other cancer cell lines with minimal toxicity to normal cells. DETA-NONOate, a long acting NO donor which has a half-life of 20 h at 37 degrees C, was used in this study. DETA-NONOate (1 mM), which releases NO in the range produced by activated macrophages, induced apoptosis after 36 h in MDA-MB-468 cells via cytochrome c release and caspase-9 and -3 activation. FTI (25 microM) potentiated the action of lower concentrations of DETA-NONOate (25-100 microM) by inducing apoptosis in these cells within 24 h by increasing cytochrome c release and caspase-9 and -3 activation. This effect was observed preferentially in the cancer cell lines studied with no apoptosis induction in normal breast epithelial cells. This novel combination of FTI and NO may emerge as a promising approach for the treatment of breast cancer. PMID- 11406541 TI - Cytokine and cyclooxygenase-2 protein in brain areas of tumor-bearing mice with prostanoid-related anorexia. AB - Evidence suggests that cytokines in the central nervous system are mediators behind anorexia in tumor-bearing hosts. We have therefore evaluated, by immunohistochemical image analyses, time course changes of interleukin (IL) 1beta, IL-6, tumor necrosis factor (TNF) alpha, IL-6 receptor (gp130), IL-1 receptor I, and cyclooxygenase (Cox)-2 protein in brain cortex, hippocampus and the ventromedial hypothalamic nucleus (VMH) in tumor-bearing mice with prostanoid related anorexia. Pair-fed non-tumor-bearing mice were used as controls. Prostaglandin E(2) was provided systemically to freely fed, non-tumor-bearing mice to confirm a role for prostanoids in modulation of brain cytokines and food intake. Time course changes of IL-1beta were significantly different between tumor-bearing mice and pair-fed controls in the hippocampus but not in the VMH. TNF-alpha in the hippocampus and VMH did not show any significant difference between tumor-bearing mice and pair-fed controls, whereas TNF-alpha showed a small increase over time in brain VMH. IL-6 content did not show any significant alterations among tumor-bearing and pair-fed mice but increased significantly over time in both the study and control group. Cox-2 in brain hippocampus and VMH showed a statistically significant rise in both tumor-bearing and pair-fed controls, with no difference between animal groups. Systemic provision of exogenous PGE(2) to non-tumor-bearing mice altered brain cytokines significantly in the hippocampus and VMH with associated changes in food intake. Our results demonstrate that some differences (IL-1beta) occurred in brain cytokines comparing tumor-bearing and pair-fed, non-tumor-bearing mice but within unexpected decreased levels in brain tissue from tumor-bearing mice. Surprisingly, many time course changes in brain cytokines were similarly altered in tumor-bearing and pair-fed mice. Our observations do not support that up regulation of brain cytokines explains or promotes anorexia in cancer disease. Rather, cytokine and Cox-dependent alterations in brain tissue seemed to be secondary to a decline in food intake and related to subsequent stress hormone activities. PMID- 11406542 TI - Eradication of colorectal xenografts by combined radioimmunotherapy and combretastatin a-4 3-O-phosphate. AB - Solid tumors have a heterogeneous pathophysiology, which has a major impact on therapy. Using SW1222 colorectal xenografts grown in nude mice, we have shown that antibody-targeted radioimmunotherapy (RIT) effectively treated the well perfused tumor rim, producing regressions for approximately 35 days, but was less effective at the more hypoxic center. By 72 h after RIT, the number of apoptotic cells rose from an overall value of 1% in untreated tumors to 35% at the tumor periphery and 10% at the center. The antivascular agent disodium combretastatin A 4 3-O-phosphate (CA4-P) rapidly reduced tumor blood flow to 62% of control values by 1 h, 23% by 3 h, and between 32-36% from 6 to 24 h after administration. This created central hemorrhagic necrosis, but a peripheral rim of cells continued to grow, and survival was unaffected. Changes in the pattern of perfusion across the tumor over time were zonal. Untreated mice showed perfusion throughout the tumor, with greatest activity at the rim. There was an overall reduction at 1 h, and total cessation of central perfusion from 3 h onward. A narrow peripheral rim of perfusion was always present, which increased in intensity and extent between 6 and 24 h, either through reperfusion or new vessel growth. Combining these two complementary therapies (7.4 MBq (131)I-labeled anti-carcinoembryonic antigen IgG i.v. plus a single 200 mg/kg dose of CA4-P i.p.) produced complete cures in five of six mice for >9 months. Allowing maximal tumor localization of antibody (48 h) before blood flow inhibition by CA4-P increased tumor retention by two to three times control levels by 96 h without altering normal tissue levels, as confirmed by gamma counting and phosphor image analysis. The success of this combined, synergistic therapy was probably the result of several factors: (a) the killing of tumor cells in the outer, radiosensitive region by targeted radiotherapy; (b) enhancement of RIT by entrapment of additional radioantibody after combretastatin induced vessel collapse; and (c) destruction of the central, more hypoxic and radioresistant region by CA4-P. This work demonstrates the need to consider cancer treatment in a biologically heterogeneous setting, if results are to be effectively translated to the clinic. PMID- 11406543 TI - Apoptosis induction in cancer cells by a novel analogue of 6-[3-(1-adamantyl)-4 hydroxyphenyl]-2-naphthalenecarboxylic acid lacking retinoid receptor transcriptional activation activity. AB - The retinoid 6-[3-(1-adamantyl)-4-hydroxyphenyl]-2-naphthalenecarboxylic acid (AHPN) is reported to have anticancer activity in vivo. Induction of cell cycle arrest and apoptosis in cancer cell lines refractory to standard retinoids suggests a retinoid-independent mechanism of action for AHPN. Conformational studies suggested that binding of AHPN does not induce an unusual conformation in retinoic acid receptor (RAR) gamma. The 3-chloro AHPN analogue MM11453 inhibited the growth of both retinoid-resistant (HL-60R leukemia, MDA-MB-231 breast, and H292 lung) and retinoid-sensitive (MCF-7 breast, LNCaP prostate, and H460 lung) cancer cell lines by inducing apoptosis at similar concentrations. Before apoptosis, MM11453 induced transcription factor TR3 expression and loss of mitochondrial membrane potential characteristic of apoptosis. MM11453 lacked the ability to significantly activate RARs and retinoid X receptor alpha to initiate (TREpal)(2)-tk-CAT reporter transcription. These results, differential proteolysis-sensitivity assays, and glutathione S-transferase-pulldown experiments demonstrate that, unlike AHPN or the natural or standard synthetic retinoids, MM11453 does not behave as a RAR or retinoid X receptor alpha transcriptional agonist. These studies strongly suggest that AHPN exerts its cell cycle arrest and apoptotic activity by a signaling pathway independent of retinoid receptor activation. PMID- 11406544 TI - Resveratrol induces extensive apoptosis by depolarizing mitochondrial membranes and activating caspase-9 in acute lymphoblastic leukemia cells. AB - Resveratrol, a plant antibiotic, has been found to have anticancer activity and was recently reported to induce apoptosis in the myeloid leukemia line HL60 by the CD95-CD95 ligand pathway. However, many acute lymphoblastic leukemias (ALLs), particularly of B-lineage, are resistant to CD95-mediated apoptosis. Using leukemia lines derived from patients with pro-B t(4;11), pre-B, and T-cell ALL, we show in this report that resveratrol induces extensive apoptotic cell death not only in CD95-sensitive leukemia lines, but also in B-lineage leukemic cells that are resistant to CD95-signaling. Multiple dose treatments of the leukemic cells with 50 microM resveratrol resulted in >/=80% cell death with no statistically significant cytotoxicity against normal peripheral blood mononuclear cells under identical conditions. Resveratrol treatment did not increase CD95 expression or trigger sensitivity to CD95-mediated apoptosis in the ALL lines. Inhibition of CD95-signaling with a CD95-specific antagonistic antibody indicated that CD95-CD95 ligand interactions were not involved in initiating resveratrol-induced apoptosis. However, in each ALL line, resveratrol induced progressive loss of mitochondrial membrane potential as measured by the dual emission pattern of the mitochondria-selective dye JC-1. The broad spectrum caspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone failed to block the depolarization of mitochondrial membranes induced by resveratrol, further indicating that resveratrol action was independent of upstream caspase-8 activation via receptor ligation. However, increases in caspase-9 activity ranged from 4- to 9-fold in the eight cell lines after treatment with resveratrol. Taken together, these results point to a general mechanism of apoptosis induction by resveratrol in ALL cells that involves a mitochondria/caspase-9-specific pathway for the activation of the caspase cascade and is independent of CD95-signaling. PMID- 11406545 TI - The cell transmembrane pH gradient in tumors enhances cytotoxicity of specific weak acid chemotherapeutics. AB - The extracellular pH is lower in tumor than in normal tissue, whereas their intracellular pH is similar. In this study, we show that the tumor-specific pH gradient may be exploited for the treatment of cancer by weak acid chemotherapeutics. i.v.-injected glucose substantially decreased the electrode estimated extracellular pH in a xenografted human tumor while its intracellular pH, evaluated by (31)P magnetic resonance spectroscopy, remained virtually unchanged. The resulting increase in the average cell pH gradient caused a parallel increase in tumor growth delay by the weak acid chlorambucil (CHL). Regardless of glucose administration, the effect of CHL was significantly greater in tumors preirradiated with a large dose of ionizing radiation. This suggests that CHL was especially pronounced in radioresistant hypoxic cells possessing a larger transmembrane pH gradient. These results indicate that the naturally occurring cell pH gradient difference between tumor and normal tissue is a major and exploitable determinant of the uptake of weak acids in the complex tumor microenvironment. The use of such drugs may be especially effective in combination with radiation. PMID- 11406546 TI - Trastuzumab (herceptin), a humanized anti-Her2 receptor monoclonal antibody, inhibits basal and activated Her2 ectodomain cleavage in breast cancer cells. AB - HER2 is a ligand-less tyrosine kinase receptor of the ErbB family that is frequently overexpressed in breast cancer. It undergoes proteolytic cleavage that results in the release of the extracellular domain and the production of a truncated membrane-bound fragment, p95. We show that HER2 shedding is activated by 4-aminophenylmercuric acetate (APMA), a well-known matrix metalloprotease activator, in HER2-overexpressing breast cancer cells. The HER2 p95 fragment, which appears after APMA-induced cleavage, is phosphorylated. We analyzed 24 human breast cancer specimens, and a phosphorylated M(r) 95,000 HER2 band could be detected in some of them, which indicated that the truncated receptor is also present in vivo. The activation of HER2 shedding by APMA in cells was blocked with batimastat, a broad-spectrum metalloprotease inhibitor. Trastuzumab (Herceptin; Genentech, San Francisco, CA), a humanized monoclonal antibody directed at the HER2 ectodomain, which has been shown to be active in patients with HER2-overexpressing breast cancer, inhibited basal and induced HER2 cleavage and, as a consequence, the generation of phosphorylated p95. This inhibitory effect of trastuzumab was not shared by 2C4, an antibody against a different epitope of the HER2 ectodomain. The inhibition of basal and APMA-induced cleavage of HER2 by trastuzumab preceded antibody-induced receptor down-modulation, which indicated that the effect of trastuzumab on cleavage was not attributable to a decrease in cell-surface HER2 induced by trastuzumab. We propose that the inhibition of HER2 cleavage and prevention of the production of an active truncated HER2 fragment represent a novel mechanism of action of trastuzumab. PMID- 11406547 TI - High affinity restricts the localization and tumor penetration of single-chain fv antibody molecules. AB - Antitumor monoclonal antibodies must bind to tumor antigens with high affinity to achieve durable tumor retention. This has spurred efforts to generate high affinity antibodies for use in cancer therapy. However, it has been hypothesized that very high affinity interactions between antibodies and tumor antigens may impair efficient tumor penetration of the monoclonal antibodies and thus diminish effective in vivo targeting (K. Fujimori et al., J. Nucl. Med., 31: 1191-1198, 1990). Here we show that intrinsic affinity properties regulate the quantitative delivery of antitumor single-chain Fv (scFv) molecules to solid tumors and the penetration of scFv from the vasculature into tumor masses. In biodistribution studies examining a series of radioiodinated scFv mutants with affinities ranging from 10(-7)-10(-11) M, quantitative tumor retention did not significantly increase with enhancements in affinity beyond 10(-9) M. Similar distribution patterns were observed when the scFv were evaluated in the absence of renal clearance in anephric mice, indicating that the rapid renal clearance of the scFv was not responsible for these observations. IHC and IF evaluations of tumor sections after the i.v. administration of scFv affinity mutants revealed that the lowest affinity molecule exhibited diffuse tumor staining whereas the highest affinity scFv was primarily retained in the perivascular regions of the tumor. These results indicate that antibody-based molecules with extremely high affinity have impaired tumor penetration properties that must be considered in the design of antibody-based cancer therapies. PMID- 11406548 TI - Activated granulocytes and granulocyte-derived hydrogen peroxide are the underlying mechanism of suppression of t-cell function in advanced cancer patients. AB - Impaired T-cell function in patients with advanced cancer has been a widely acknowledged finding, but mechanisms reported thus far are those primarily operating in the tumor microenvironment. Very few mechanisms have been put forth to explain several well-described defects in peripheral blood T cells, such as reduction in expression of signaling molecules, decreased production of cytokines, or increased apoptosis. We have closely examined the peripheral blood mononuclear cell (PBMC) samples derived from patients and healthy individuals, and we have observed an important difference that may underlie the majority of reported defects. We observed that in samples from patients only, an unusually large number of granulocytes copurify with low density PBMCs on a density gradient rather than sediment, as expected, to the bottom of the gradient. We also show that activating granulocytes from a healthy donor with N-formyl-L methionyl-L-leucyl-L-phenylalanine could also cause them to sediment aberrantly and copurify with PBMCs, suggesting that density change is a marker of their activation. To confirm this, we looked for other evidence of in vivo granulocyte activation and found it in drastically elevated plasma levels of 8-isoprostane, a product of lipid peroxidation and a marker of oxidative stress. Reduced T-cell receptor zeta chain expression and decreased cytokine production by patients' T cells correlated with the presence of activated granulocytes in their PBMCs. We showed that freshly obtained granulocytes from healthy donors, if activated, can also inhibit cytokine production by T cells. This action is abrogated by the addition of the hydrogen peroxide (H(2)O(2)) scavenger, catalase, implicating H(2)O(2) as the effector molecule. Indeed, when added alone, H(2)O(2) could suppress cytokine production of normal T cells. These findings indicate that granulocytes are activated in advanced cancer patients and that granulocyte derived H(2)O(2) is the major cause of severe systemic T-cell suppression. PMID- 11406549 TI - Generation of cytotoxic T lymphocytes against native and altered peptides of human leukocyte antigen-A*0201 restricted epitopes from the human epithelial cell adhesion molecule. AB - A growing number of human tumor antigens have been described that can be recognized by CTLs in a MHC class I restricted fashion. The epithelial cell adhesion molecule (Ep-CAM) is expressed in a variety of human tumors and has attracted attention as a therapeutic target for monoclonal antibody serotherapy. We have identified immunogenic peptides derived from Ep-CAM, that bind to human leukocyte antigen-A*0201 and elicit strong peptide-specific human CTL responses, demonstrating that there is an effective T-cell repertoire against these Ep-CAM derived peptides that can be recruited. Alterations to these peptides were made to increase their binding affinity to MHC class I molecules. The use of such "heteroclitic" peptides allowed generation of cytotoxic T cells that demonstrated increased killing of target cells pulsed not only with the heteroclitic but also with the native peptide. Most important, CTL cell lines that are generated against these peptides specifically lyse epithelial tumor cells expressing Ep-CAM but not normal hematopoietic or bronchial epithelial cells. PMID- 11406550 TI - Regulatory CD4(+)CD25(+) T cells in tumors from patients with early-stage non small cell lung cancer and late-stage ovarian cancer. AB - Immunosuppression may contribute to the progression of cancer. In this study we assessed the structural and functional status of T cells from tumor specimens obtained from patients with early stage non-small cell lung cancer and late-stage ovarian cancer. Although some groups have described structural alterations in the TCR in patients with other malignancies, we did not observe decreased expression of the CD3zeta subunit in the tumor-associated T cells. However, increased percentages of CD4(+)CD25(+) T cells were observed in the non-small cell lung cancer tumor-infiltrating lymphocytes and ovarian cancer tumor-associated lymphocytes. Furthermore, these CD4(+)CD25(+) T cells were found to secrete transforming growth factor-beta, consistent with the phenotype of regulatory T cells. Despite a generalized expression of lymphocyte activation markers in the tumor-associated T-cell populations, the CD8(+) T cells expressed low levels of CD25. To determine whether expression of CD25 could be restored on the CD8 cells, tumor-associated T cells were stimulated with anti-CD3 and anti-CD28 monoclonal antibodies. After stimulation, nearly all of the CD8 T cells expressed CD25. Furthermore, despite the low levels of interleukin 2, IFN-gamma, and tumor necrosis factor-alpha secretion by the tumor-associated and peripheral blood T cells at baseline, stimulation with anti-CD3 and anti-CD28 monoclonal antibodies significantly increased the fraction of cells producing these cytokines. Thus, tumor-associated T cells from patients with early and late-stage epithelial tumors contain increased proportions of CD4(+)CD25(+) T cells that secrete the immunosuppressive cytokine transforming growth factor-beta. Furthermore, our results are consistent with previous reports showing impaired expression of CD25 on CD8(+) T cells in cancer patients. Finally, increased lymphocyte costimulation provided by triggering the CD28 receptor is able to increase CD25 expression and cytokine secretion in tumor-associated T cells. These observations provide evidence for the contribution of regulatory T cells to immune dysfunction in cancer patients. PMID- 11406551 TI - Tumor-reactive T helper lymphocytes recognize a promiscuous MAGE-A3 epitope presented by various major histocompatibility complex class II alleles. AB - The development of effective T cell-based immunotherapy for cancer requires the identification of antigens capable of inducing both CTL and T helper immune responses. Although CTLs will participate in the antitumor response mainly by exerting their lytic activity on the tumor cells, helper T lymphocytes will be critical for the induction and maintenance of the CTLs. Thus, effective subunit therapeutic vaccines should include both CTL and T helper epitopes from antigens expressed on the tumor cells. The product of the MAGE-A3 gene is an attractive candidate for tumor immunotherapy because it is expressed in the majority of melanomas and in a great proportion of other solid tumors. Although numerous CTL epitopes for the MAGE-A3 antigen have been reported, only a few have been described for helper T cells. Here we show that a synthetic peptide derived from the MAGE-A3 sequence (MAGE-A3(146-160)) was effective in inducing in vitro T helper responses in the context of HLA-DR4 and HLA-DR7 alleles. Most significantly, the peptide-reactive helper T lymphocytes were capable of recognizing various forms of MAGE-A3 antigen (tumor cell lysates, dead/apoptotic tumor cells, or recombinant MAGE-A3 protein), indicating that the T-cell epitope represented by peptide MAGE-A3(146-160) is naturally processed by antigen presenting cells. These studies are relevant for the design of multi-epitope vaccines for treating MAGE-A3-expressing tumors through the simultaneous stimulation of CTL and T helper lymphocytes. PMID- 11406552 TI - Transduction of antisense cyclin D1 using two-step gene transfer inhibits the growth of rat hepatoma cells. AB - Cyclin D1, one of the G(1) cyclins, is frequently overexpressed in several types of carcinomas and is thought to play an important role in tumorigenesis and tumor progression including hepatocellular carcinoma. We constructed a retrovirus vector-carrying rat cyclin D1 cDNA in the reverse orientation, resulting in expression of antisense (AS) cyclin D1 mRNA. For efficient transduction of this recombinant retrovirus, two-step gene transfer was performed. The rat hepatoma cell line (dRLh84) was infected with this recombinant retrovirus after preinfection with adenovirus expressing the retrovirus receptor. In the rat hepatoma cells, AS cyclin D1 mRNA was expressed, inducing a decrease in the expression of endogenous cyclin D1 mRNA and an inhibition of cell growth. Moreover, two-step gene transfer of AS cyclin D1 into s.c. hepatoma xenografts resulted in inhibition of tumor growth and prolonged animal survival. In the virus-infected tumor xenografts, expression of cyclin D1 was immunohistochemically inhibited, and apoptosis of hepatoma cells was detected. These findings suggest that transduction of AS cyclin D1 is useful as an adjunct to standard treatments for hepatocellular carcinoma. PMID- 11406553 TI - Limitations of the reporter green fluorescent protein under simulated tumor conditions. AB - This paper reports a detailed analysis of the effect of low oxygen conditions (hypoxia) on the reporter green fluorescent protein (GFP). It questions the feasibility of using GFP for gene expression studies under tumor conditions. Hypoxia is a characteristic of both experimental and clinical tumors. Several important factors are pointed out which need to be considered when using GFP as reporter gene. GFP fluorescence is the final product of a long and complex pathway involving transcription, translation, and posttranslational modifications. All of these steps may be affected by the availability of oxygen. We show specifically that cellular GFP fluorescence decreased with reduced oxygenation, anoxia virtually eliminated fluorescence and protein levels, and fluorescence recovery after anoxia required 5-10 h of reoxygenation. In conclusion, GFP appears to be a good marker gene to study location or movement of proteins or cells but should be used with great caution as a reporter of gene expression under tumor conditions. PMID- 11406554 TI - A novel mechanism for chaperone-mediated telomerase regulation during prostate cancer progression. AB - Telomerase activity has been detected in >85% of all malignant human cancers, including 90% of prostate carcinomas. Using a well-characterized experimental prostate cancer system, we have found that telomerase activity is notably increased (>10-fold) during tumorigenic conversion. Expression profiles of the telomerase components (hTR and hTERT) revealed no substantive changes, which suggests a nontranscriptional mechanism for increased activity. Because the hsp90 chaperone complex functionally associates with telomerase, we investigated that relationship and found that along with telomerase activity, a number of hsp90 related chaperones are markedly elevated during transformation, as well as in advanced prostate carcinomas. Using the nontumorigenic cell protein extract as the source of telomerase, addition of purified chaperone components enhanced reconstitution of telomerase activity, which suggests a novel mechanism of increased telomerase assembly via a hsp90 chaperoning process during prostate cancer progression. PMID- 11406555 TI - Molecular profiling of transformed and metastatic murine squamous carcinoma cells by differential display and cDNA microarray reveals altered expression of multiple genes related to growth, apoptosis, angiogenesis, and the NF-kappaB signal pathway. AB - To identify changes in gene expression with transformation and metastasis, we investigated differential gene expression in a squamous carcinoma model established in syngeneic mice. We used mRNA differential display (DD) to detect global differences and cDNA arrays enriched for cancer-associated genes using mRNA from primary keratinocytes, transformed Pam 212 squamous carcinoma cells, and metastases of Pam 212. After DD, 72 candidate cDNAs expressed primarily in transformed and metastatic cells were selected and cloned. Fifty-seven were detected, and 32 were confirmed to be differentially expressed by Northern blot analysis. mRNA expression profiles were also generated using a mouse cDNA array composed of 4000 elements representing known genes and expressed sequence tags plus the 57 DD candidate cDNAs detected by Northern analysis to facilitate data validation. cDNA array detected 76.9% of the differentially expressed mRNAs selected from DD and confirmed by Northern blot, whereas low-abundance mRNAs did not reach the threshold for detection by the lower-sensitivity array method. Clustering analysis of DD and array results from transformed and metastatic cells identified genes that exhibited decreased or increased expression with transformation and metastasis. Alterations in the expression of several genes detected during tumor progression were consistent with their functional activities involving growth (p21, p27, and cyclin D1), resistance and apoptosis (glutathione-S-transferase, cIAP-1, PEA-15, and Fas ligand), inflammation and angiogenesis [chemokine growth-regulated oncogene 1 (also called KC)], and signal transduction (c-Met, yes-associated protein, and syk). Strikingly, 10 of 22 genes in the cluster expressed in metastases have been associated with activation of the nuclear factor (NF)-kappaB signal pathway. The NF-kappaB-inducible cytokine Gro-1 was recently shown to promote tumor growth, metastasis, and angiogenesis of squamous cell carcinomas in vivo (Loukinova et al., Oncogene, 19: 3477-3486, 2000). The results demonstrate that early response genes related to NF-kappaB contribute to metastatic tumor progression. Comparison of cell lines and tumor tissue revealed a concordance of approximately 50% by array, and 70% for Northern confirmed, metastasis-related genes. Functional genomic approaches comparing expression among cell lines and tumor tissue may promote a better understanding of the genes expressed by malignant and host cells during tumor progression and metastasis. PMID- 11406556 TI - MAGE-E1, a new member of the melanoma-associated antigen gene family and its expression in human glioma. AB - To unearth glioma-specific genes in human glioblastoma, the serial analysis of gene expression technique was applied to a primary glioblastoma, using cultured human astrocytes as a normal control. Among the top 147 most-expressed tags in glioblastoma, we found a tag, TTTTGGGTAT, that originated from an unidentified gene and which was not detected in human astrocyte cultures. Real-time quantitative reverse transcription-PCR showed that MAGE-E1 expression was 2.6-15 fold enriched in glioblastoma relative to human astrocytes. Expressed sequence tags containing this tag were homologous to the melanoma-associated antigen gene (MAGE) family, and this new cDNA, named MAGE-E1, was cloned by the 5'-rapid amplification of cDNA ends technique. Three alternatively spliced variants (MAGE E1a-c) were found, and deduced amino acid sequence showed that MAGE-E1a and -E1b shared the MAGE-conserved region, whereas -E1c did not. This suggests that although MAGE-E1c is expressed from one of the MAGE family, it has distinct functions from other members. Tissue distribution analysis showed that MAGE-E1 was distinct from other MAGEs. MAGE-E1 expression was detected only in brain and ovary among normal tissues. Interestingly, MAGE-E1a and/or -E1b were specifically expressed in glioma cells among cancer cells. These results indicate that MAGE-E1 is a novel and glioma-specific member of MAGE family. PMID- 11406557 TI - VHL and FHIT locus loss of heterozygosity is common in all renal cancer morphotypes but differs in pattern and prognostic significance. AB - Deletions involving 3p are believed to be typical for conventional (clear cell) renal cell carcinoma (cRCC), with confirmed and suspected targets being the VHL and FHIT tumor suppressor genes, respectively. By contrast, 3p deletions are felt to be rare in papillary RCC (pRCC) and chromophobe RCC (chRCC); however, this belief is based on relatively scant data. In particular, 3p14.2 deletions, possibly resulting in FHIT inactivation, have been rarely studied in pRCC or chRCC even though they may be relevant in early renal tumorigenesis. We therefore examined 3p deletion rates and patterns in pRCC and chRCC with particular attention to 3p14.2. We examined 16 chRCCs and 27 pRCCs for loss of heterozygosity (LOH) at 3p25-26 and 3p14.2 using 13 well-mapped microsatellite markers. Those pRCC with LOH at 3p25-26 were also screened for VHL gene mutations. The results were correlated with tumor histology and patient outcome and compared with data we had obtained previously on cRCC. We found similar overall 3p LOH rates in pRCC (59%), chRCC (86.6%), and cRCC (75.8%). In pRCC and chRCC, LOH at 3p25-26 was more common than at 3p14.2, whereas the converse was true for cRCC. In the pRCC with 3p25-26 LOH, we confirmed that this was not associated with mutations of the VHL gene. At 3p14.2, LOH rates of pRCC were lower than those of cRCC and chRCC (p<0.02). All morphotypes showed a predominately interstitial LOH pattern, which was most pronounced in the 3p14.2 region in cRCC. 3p LOH in chRCC was associated with improved patient outcome, mirroring our previous cRCC data. We conclude that 3p LOH is a universal phenomenon in RCC, but has different underlying mechanisms, molecular targets, and implications in the different morphotypes, although FHIT inactivation may play a role in both cRCC and chRCC tumorigenesis. PMID- 11406558 TI - Cytosine methylation represses glutathione S-transferase P1 (GSTP1) gene expression in human prostate cancer cells. AB - Methylation of the glutathione S-transferase P1 (GSTP1) gene has been described as a highly specific and sensitive biomarker for prostate cancer. However, at present, it is not known whether methylation represses GSTP1 gene expression in human prostate cancer. We found the GSTP1 gene promoter to be completely methylated in the LNCaP prostate cancer cell line, where this gene is transcriptionally inactive. In contrast, Du145 and PC3 prostate cancer cells express the GSTP1 gene and exhibit methylated and unmethylated GSTP1 alleles. In a transient transfection assay using LNCaP cells, methylation of the GSTP1 promoter-driven luciferase reporter vector (GSTP1-pGL3) resulted in a >20-fold inhibition of transcription, and this repression was not relieved by the presence of a histone deacetylase inhibitor, trichostatin A (TSA). Treatment of LNCaP cells with a DNA methyltransferase inhibitor, 5-Aza-2'-deoxycytidine, resulted in demethylation and activation of the GSTP1 gene. In contrast, TSA treatment failed to demethylate or activate the GSTP1 gene. Fully methylated but not unmethylated GSTP1 promoter fragment was shown to bind to a complex similar to methyl cytosine binding protein complex 1 that contains methyl-CpG-binding domain 2 protein (MBD2) in electrophoretic mobility shift assays using LNCaP cell nuclear extracts. These data demonstrate that cytosine methylation can repress GSTP1 gene expression in LNCaP prostate cancer cells and that this effect is possibly mediated by a methyl cytosine-binding protein complex 1-like complex. Furthermore, these data also support the notion of the dominance of methylation over TSA-sensitive histone deacetylation in silencing genes with a high CpG density in the promoter region. PMID- 11406559 TI - Fragile histidine triad expression delays tumor development and induces apoptosis in human pancreatic cancer. AB - The fragile histidine triad (FHIT) gene is a tumor suppressor gene that is altered by deletion in a large fraction of human tumors, including pancreatic cancer. To evaluate the potential of FHIT gene therapy, we developed recombinant adenoviral and adenoassociated viral (AAV) FHIT vectors and tested these vectors in vitro and in vivo for activity against human pancreatic cancer cells. Our data show that viral FHIT gene delivery results in apoptosis by activation of the caspase pathway. Furthermore, Fhit overexpression enhances the susceptibility of pancreatic cancer cells to exogenous inducers of apoptosis. In vivo results show that FHIT gene transfer delays tumor growth and prolongs survival in a murine model mimicking human disease. PMID- 11406560 TI - Induction of anoikis and suppression of human ovarian tumor growth in vivo by down-regulation of Bcl-X(L). AB - Normal or immortal epithelial cells are sensitive to a form of apoptosis, commonly referred to as anoikis, which is induced by detachment from the extracellular matrix (ECM). In contrast, development of carcinomas is associated with acquisition of cellular resistance to anoikis. However, whether human cancer cells deprived of anoikis resistance necessarily display reduced tumorigenic properties in vivo is unknown. We decided to address this question using human ovarian carcinoma cells as a model. Bcl-X(L), an apoptotic factor considered to play an important role in (resistance to) anoikis, is overexpressed in ovarian cancer, and represents an unfavorable prognostic indicator for this type of human malignancy. We therefore evaluated whether Bcl-X(L) can be used as a tool to manipulate anoikis resistance and tumorigenicity of ovarian cancer cells. We show here that when nonmalignant ovarian epithelial cells are detached from the ECM, down-regulation of Bcl-X(L) and apoptotic cell death are observed, although these events do not occur in ovarian carcinoma cells. Moreover, enforced down regulation of Bcl-X(L) by transfection with antisense cDNA in the anoikis resistant and highly tumorigenic HEY ovarian carcinoma cell line had no impact on the viability of these cells under adherent conditions but caused significant apoptosis in response to detachment from the ECM. This change was associated with a strong inhibition of tumorigenicity of the Bcl-X(L)-deficient HEY cells in nude mice, both s.c. and in the peritoneal cavity. These results suggest a critical role for Bcl-X(L) in the maintenance of anoikis resistance in ovarian cancer cells. They also serve to establish a functional linkage between this property and the ability of human cancer cells to grow aggressively in vivo. Consequently, targeting molecular mechanisms responsible for anoikis resistance may serve as a potentially effective therapeutic strategy for the treatment of such human malignancies as ovarian cancer. PMID- 11406561 TI - Homology-directed dna repair, mitomycin-c resistance, and chromosome stability is restored with correction of a Brca1 mutation. AB - Chromosomal breaks occur spontaneously as a result of normal DNA metabolism and after exposure to DNA-damaging agents. A major pathway involved in chromosomal double-strand break repair is homologous recombination. In this pathway, a DNA sequence with similarity to a damaged chromosome directs the repair of the damage. The protein products of the hereditary breast cancer susceptibility genes, BRCA1 and BRCA2, interact with the Rad51 protein, a central component of homologous repair pathways. We have recently shown that this interaction is significant by demonstrating that Brca1- and BRCA2-deficient cells are defective in homology-directed chromosomal break repair. We confirm that Brca1-deficient embryonic stem (ES) cells are defective in gene targeting and homology-directed repair of an I-Sce I-induced chromosome break. The phenotypic paradigm that defines homology-directed repair mutants is extended to these Brca1-deficient cells by the demonstration of 100-fold sensitivity to the interstrand cross linking agent mitomycin-C and spontaneous chromosome instability. Interestingly, although chromosome aberrations were evident, aneuploidy was not observed. Repair phenotypes are partially restored by expression of a Brca1 transgene, whereas correction of one mutated Brca1 allele through gene targeting fully restores mitomycin-C resistance and chromosome stability. We conclude that the inability to properly repair strand breaks by homology-directed repair gives rise to defects in chromosome maintenance that promote genetic instability and, it is likely, tumorigenesis. PMID- 11406562 TI - The role of Thomsen-Friedenreich antigen in adhesion of human breast and prostate cancer cells to the endothelium. AB - Interactions of metastatic cancer cells with vasculatory endothelium are critical during early stages of cancer metastasis. Understanding the molecular underpinnings of these interactions is essential for the development of new efficacious cancer therapies. Here we demonstrate that cancer-associated carbohydrate T antigen plays a leading role in docking breast and prostate cancer cells onto endothelium by specifically interacting with endothelium-expressed beta-galactoside-binding protein, galectin-3. Importantly, T antigen-bearing glycoproteins are also capable of mobilizing galectin-3 to the surface of endothelial cells, thus priming them for harboring metastatic cancer cells. The T antigen-mediated, tumor-endothelial cell interactions could be efficiently disrupted using synthetic compounds either mimicking or masking this carbohydrate structure. High efficiency of T antigen-mimicking and T antigen-masking inhibitors of tumor cell adhesion warrants their further development into antiadhesive cancer therapeutics. PMID- 11406563 TI - Tetrasomy is induced by human papillomavirus type 18 E7 gene expression in keratinocyte raft cultures. AB - We have demonstrated previously that oncogenic human papillomaviruses (HPVs) induce basal cell tetrasomy in low-grade squamous intraepithelial lesions of the cervix. To identify HPV genes and growth conditions involved in this process, we analyzed: (a) organotypic raft cultures of primary human keratinocytes transfected with whole HPV-18 genomes; and (b) organotypic raft cultures acutely infected with recombinant retroviruses expressing the HPV-18 E6, E7, or E6/E7 genes from the differentiation-dependent HPV-18 enhancer-promoter. Cultures were examined for HPV DNA by in situ hybridization and for karyotype by interphase cytogenetics. Tetrasomy occurred in the suprabasal strata of raft cultures expressing E7 and E6/E7 but not in those expressing E6 alone or in a control culture. These data indicate that suprabasal tetrasomy occurs in association with expression of the E7 gene alone. Basal cell tetrasomy was additionally observed in the raft culture transfected with whole HPV-18 genomes, consistent with observations in low-grade squamous intraepithelial lesions. The distribution of tetrasomic cells in these raft cultures may reflect the involvement of additional viral genes or possibly differences in the pattern of viral oncogene and host gene expression. PMID- 11406564 TI - Fas-mediated apoptosis in neuroblastoma requires mitochondrial activation and is inhibited by FLICE inhibitor protein and Bcl-2. AB - Fas-mediated apoptosis proceeds though mitochondria-dependent or -independent pathways and is deficient in drug-resistant cells. Neuroblastoma, a common pediatric malignancy, often develops drug-resistance and has a silenced caspase 8 (FLICE) gene, which has been associated with Fas- and drug-resistance. We report that besides caspase 8, which was absent in approximately one-third of 26 neuroblastoma cases in this study, other proteins such as bcl-2 and FLICE inhibitory protein (FLIP), are equally important in conferring Fas-resistance to neuroblastoma cells. Both bcl-2 and FLIP were frequently expressed in neuroblastoma tissues. Our in vitro studies showed that FLIP was recruited to the death-inducing signaling complex and interfered with the recruitment of caspase 8 in neuroblastoma cells. bcl-2 inhibited the activation of the mitochondria; but it also lowered the free cytoplasmic levels of caspase 8 by binding and sequestering it, thus acting through a novel antiapoptotic mechanism upstream of the mitochondria. In vitro down-regulation of bcl-2 with antisense oligonucleotides allowed the release of cytochrome c from mitochondria and the activation of caspases 8 and 3 upon Fas activation as well as sensitized neuroblastoma cells to Fas-mediated apoptosis. Down-regulation of FLIP had only a modest apoptotic effect because of the coexistent mitochondrial block. However, combined treatment with bcl-2 and FLIP antisense oligonucleotides had a statistically significant synergistic effect reversing Fas-resistance in neuroblastoma cells in vitro. These data indicate that Fas-mediated apoptosis in neuroblastoma cells is mitochondria-dependent and inhibited both at the mitochondrial level and at the level of caspase 8 activation. Thus, gene targeting therapies for bcl-2 and FLIP may reverse Fas-resistance and prove useful in the treatment of drug-resistant neuroblastomas. PMID- 11406565 TI - In vivo molecular imaging of met tyrosine kinase growth factor receptor activity in normal organs and breast tumors. AB - Molecular imaging techniques allow visualization of specific gene products and their physiological processes in living tissues. In this study, we present a new approach for molecular imaging of endogenous tyrosine kinase receptor activity. Met and its ligand hepatocyte growth factor scatter factor (HGF/SF), which mediate mitogenicity, tumorigenicity, and angiogenesis, were used as a model. HGF/SF and Met play a significant role in the pathogenesis and biology of a wide variety of human epithelial cancers and, therefore, may serve as potential targets for cancer prognosis and therapy. We have shown previously that in vitro activation of Met by HGF/SF increases oxygen consumption. In this study, we demonstrate that Met activation in vivo by HGF/SF alters the hemodynamics of normal and malignant Met-expressing tissues. Tumor-bearing BALB/C mice were i.v. injected with HGF/SF and imaged using magnetic resonance imaging (MRI) and Doppler ultrasound. Organs and tumors expressing high levels of Met showed the most substantial alteration in blood oxygenation levels as measured by blood oxygenation level depended (BOLD)-MRI. No significant alteration was observed in tumors or organs that does not express Met. In the liver, which expresses high levels of Met, MRI signal alteration of about 60% was observed. In the kidneys, signal alteration was approximately 30%, and no change was observed in muscles. The extent of MRI signal alteration was also in correlation with HGF/SF doses. Injection of 7 and 170 ng/g body weight resulted in signal alteration of 5% and 30%, respectively, in tumors. Doppler ultrasound measurements demonstrated that these MRI changes are at least partially attributable to altered blood flow. These hemodynamic alterations, measured by MRI and Doppler ultrasound, were used in this study for the molecular imaging of Met activity in vivo. This novel molecular imaging technique may be used for in vivo diagnosis, prognosis, and therapy of Met-expressing tumors. PMID- 11406566 TI - Nonsteroidal anti-inflammatory drugs induce apoptosis in esophageal cancer cells by restoring 15-lipoxygenase-1 expression. AB - In previous studies, we have found that expression of 15-lipoxygenase-1 (15-LOX 1) and its main product, 13-S-hydroxyoctadecadienoic acid, are decreased in human colorectal cancers and that nonsteroidal anti-inflammatory drugs (NSAIDs) can therapeutically induce 15-LOX-1 expression to trigger apoptosis in human colorectal cancer cells. NSAIDs similarly induce apoptosis in esophageal cancer cells, although the mechanisms of these effects remain to be defined. In the present study, we tested whether 15-LOX-1 is down-regulated in human esophageal cancers using paired normal and tumor human surgical samples and whether NSAIDs can up-regulate 15-LOX-1 to restore apoptosis in esophageal cancer cells. We found that: (a) 15-LOX-1 was down-regulated in human esophageal carcinomas; (b) NSAIDs induced 15-LOX-1 expression during apoptosis in esophageal cancer cells; and (c) 15-LOX-1 inhibition suppressed NSAID-induced apoptosis, which was restored by 13-S-hydroxyoctadecadienoic acid but not by its parent compound, linoleic acid. These findings demonstrate that 15-LOX-1 is down-regulated in human esophageal carcinomas and that NSAIDs induce apoptosis in esophageal cancer cells via up-regulation of 15-LOX-1. They also support the concept that the loss of the proapoptotic role of 15-LOX-1 in epithelial cancers is not limited to human colorectal cancers. PMID- 11406567 TI - Inhibition of epidermal growth factor-induced RhoA translocation and invasion of human pancreatic cancer cells by 3-hydroxy-3-methylglutaryl-coenzyme a reductase inhibitors. AB - 3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitors prevent the conversion of HMG-CoA to mevalonate and thereby inhibit the synthesis of other products derived from this metabolite. This includes a number of small prenylated GTPases involved in cell growth, motility, and invasion. We studied the effect of HMG-CoA reductase inhibitors (fluvastatin and lovastatin) on in vitro invasion of human pancreatic cancer PANC-1 cells. Epidermal growth factor (EGF) induced a dose-dependent increase of PANC-1 cell invasion in a modified Boyden chamber assay. Stimulation of cancer cells with EGF induced translocation of RhoA from the cytosol to the membrane fraction and actin stress fiber assembly. Furthermore, Clostridium botulinum C3 transferase, a specific inhibitor of Rho, inhibited the ability of EGF to promote invasion, indicating that EGF-induced cancer cell invasion is regulated by Rho signaling. Treatment of PANC-1 cells with fluvastatin markedly attenuated EGF-induced translocation of RhoA from the cytosol to the membrane fraction and actin stress fiber assembly, whereas it did not inhibit the tyrosine phosphorylation of EGF receptor and c-erbB-2. The induction of cancer cell invasion by EGF was inhibited by the addition of fluvastatin or lovastatin in a dose-dependent manner. The effects of fluvastatin or lovastatin on cell morphology and invasion were reversed by the addition of all-trans-geranylgeraniol but not by the addition of all-trans-farnesol. These results suggest that HMG-CoA reductase inhibitors affect RhoA activation by preventing geranylgeranylation, which results in inhibition of EGF-induced invasiveness of human pancreatic cancer cells. PMID- 11406568 TI - Down-regulation of the erbB-2 receptor by trastuzumab (herceptin) enhances tumor necrosis factor-related apoptosis-inducing ligand-mediated apoptosis in breast and ovarian cancer cell lines that overexpress erbB-2. AB - We investigated whether combined treatment with tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and trastuzumab could enhance the specific killing of cells that overexpress the erbB-2 receptor. The combination resulted in an enhancement of TRAIL-mediated apoptosis in all cell lines overexpressing erbB-2 receptor compared with either reagent alone. In contrast, there was no effect in cell lines with low levels of the erb-B2 receptor. Trastuzumab treatment resulted in down-regulation of the erbB-2 receptor in all erbB-2 overexpressing cell lines. Similar enhancement of TRAIL toxicity was observed when the erbB-2 receptor was down-regulated using antisense oligodeoxynucleotides. Down-regulation of the erbB-2 receptor protein by trastuzumab or antisense oligodeoxynucleotides decreased Akt kinase activation but not mitogen-activated protein kinase activation. Down-regulation of Akt kinase activity by a phosphatidylinositol 3'-kinase inhibitor (LY294002) also resulted in enhancement of TRAIL-mediated apoptosis. Expression of a constitutively active form of Akt kinase in an erbB-2-overexpressing cell line completely abrogated the increase in TRAIL-mediated apoptosis by trastuzumab and significantly reduced the biological effect of either reagent alone. Therefore, down-regulation of the erbB-2 receptor by trastuzumab enhances TRAIL-mediated apoptosis by inhibiting Akt kinase activity. These data suggest that the combination of trastuzumab and TRAIL may allow enhanced therapeutic efficacy and specificity in the treatment of erbB-2-overexpressing tumors. PMID- 11406569 TI - Constitutive IkappaB kinase activity correlates with nuclear factor-kappaB activation in human melanoma cells. AB - Constitutive IKK activity associated with increased IkappaBalpha phosphorylation and degradation contribute to the high level of endogenous nuclear factor-kappaB (NF-kappaB) activation in Hs294T melanoma cells as compared with RPE cells (R. L. Shattuck-Brandt and A. Richmond, Cancer Res., 57: 3032-3039, 1997; M. N. Devalaraja et al., Cancer Res., 59: 1372-1377, 1999). To determine whether this endogenous NF-kappaB activation was characteristic of melanoma, we examined the level of constitutive activation of NF-kappaB in a number of melanoma cell lines. We demonstrate here that eight melanoma cell lines exhibit increased IkappaB kinase (IKK) activity, enhanced phosphorylation of IkappaBalpha and p65, and enhanced nuclear localization of p65/p50 in comparison to normal human epidermal melanocytes. The chemokines, CXC ligand 1 (CXCL1) and CXCL8, but not CXCL5, are highly expressed in most of the melanoma cell lines, suggesting that the constitutive production of chemokines is highly correlated to endogenous NF kappaB activity. Our failure to observe a direct relationship between the fold activation of IKK, CXCL1, or CXCL8 mRNA levels and secretion of these chemokines into the culture medium suggest that regulation of chemokine expression also occurs at the posttranscription level of mRNA stability and/or translational control. Moreover, recombinant CXCL1 can directly induce IKK activity in normal human epidermal melanocytes in a concentration-dependent manner after up modulation of CXCL1 protein expression, whereas inhibition of IKKbeta activity results in down-modulation of CXCL1 protein expression. Finally, CXCL1 antibody blocks IKK activity and inhibits the proliferation of melanoma cells to further support the concept that the constitutive activation of NF-kappaB and autocrine effects of CXCL1 play an important role in the pathogenesis of melanoma. PMID- 11406570 TI - c-myc/p53 interaction determines sensitivity of human colon carcinoma cells to 5 fluorouracil in vitro and in vivo. AB - Colon carcinoma cells overexpress c-myc due to defective Wnt signaling, but only patients whose tumors have an amplified c-myc gene show improved disease-free and overall survival in response to 5-fluoruracil (5FU). Here we show that in two colon carcinoma cell lines that do not have an amplified c-myc gene but differ in their p53 status, high c-myc levels can be further elevated by introducing a c myc expression vector. Whereas sensitivity to low serum-induced apoptosis was imposed on the parental lines independent of p53 status and was unaffected by further elevation of c-myc, sensitivity to 5FU-induced apoptosis was dependent on both the higher c-myc levels due to the expression vector and wild-type p53 function. The elevated c-myc levels led to higher c-myc transactivation activity in the p53 wild-type cell line, but not in the mutant p53 cell line. The requirement for both elevated c-myc and p53 for 5FU sensitivity was confirmed using antisense c-myc and pifithrin-alpha, a specific inhibitor of p53. Finally, the in vitro data predicted that only patients with both amplified c-myc and wild type p53 in their primary tumors would be responsive to 5FU-based therapy, which was borne out by analysis of tumors from 135 patients entered into a Phase III clinical trial of 5FU-based adjuvant therapy. The data provide significant insight into mechanisms that establish colon tumor cell sensitivity to 5FU, clearly demonstrate the necessity of exercising caution in considering combining novel strategies that target elevated c-myc with standard 5FU-based therapy, and suggest alternative therapeutic strategies that target c-myc and/or p53 mutations in the treatment of colon cancer. PMID- 11406571 TI - A novel N-ras mutation in malignant melanoma is associated with excellent prognosis. AB - Mutations in the ras gene are key events in the process of carcinogenesis; in particular, point mutations in codon 61 of exon 2 of the N-ras gene occur frequently in malignant melanoma (MM). We searched for point mutations in the N ras gene in a large series of primary and metastatic MM from 81 different retrospectively selected patients using the very sensitive denaturing gradient gel electrophoresis technique, followed by sequencing. The classical codon 12 and codon 61 mutations were found in 21 and 17% of the cases, respectively. No codon 13 mutation was found. A novel mutation at codon 18 of exon 1, consisting of a substitution of alanine (GCA) by threonine (ACA), was found in 15% of the primary MMs but in none of the metastatic MMs. All of the other cases were free of mutations. Using microdissected cells from distinctive MM growth phases as source of DNA for mutation analysis, this particular N-ras exon 1 mutation at codon 18 was already present in the radial growth phase and preserved throughout the successive growth phases; it was also found in a dysplastic nevi in continuity with a MM, indicating a clonal relationship between both lesions. Our findings also illustrate the clonal relationship between the distinctive growth phases in MM and suggest the codon 18 mutation to occur early in MM development. The MM in patients with this mutation were significantly thinner than those without a codon 18 mutation (P = 0.0257). Statistical analysis, comparing the group of codon 18 patients with the group of patients with the classical mutations and without mutations, revealed a highly significant difference in overall outcome. The cumulative probability of developing metastasis was significantly lower for the group patients with a codon 18 mutation (P = 0.0130). We can thus conclude that this codon 18 mutation identifies a group of patients with better prognosis than patients with melanoma that harbor wild-type sequence or classical activating point mutations in codon 12 or 61. Preliminary nucleotide binding measurements could not detect a difference between wild-type Ras protein and the mutant Ras(A18T) protein. However, for a precise elucidation of the role of the N Ras(A18T) mutant in melanoma, additional studies aimed to measure the affinity to guanine nucleotide exchange factors and GTPase-activating proteins are needed. PMID- 11406572 TI - A white paper: the product of a pancreas cancer think tank. PMID- 11406575 TI - The inner nuclear membrane: simple, or very complex? PMID- 11406576 TI - Novel recognition mode between Vav and Grb2 SH3 domains. AB - Vav is a guanine nucleotide exchange factor for the Rho/Rac family that is expressed exclusively in hematopoietic cells. Growth factor receptor-bound protein 2 (Grb2) has been proposed to play important roles in the membrane localization and activation of Vav through dimerization of its C-terminal Src homology 3 (SH3) domain (GrbS) and the N-terminal SH3 domain of Vav (VavS). The crystal structure of VavS complexed with GrbS has been solved. VavS is distinct from other SH3 domain proteins in that its binding site for proline-rich peptides is blocked by its own RT loop. One of the ends of the VavS beta-barrel forms a concave hydrophobic surface. The GrbS components make a contiguous complementary interface with the VavS surface. The binding site of GrbS for VavS partially overlaps with the canonical binding site for proline-rich peptides, but is definitely different. Mutations at the interface caused a decrease in the binding affinity of VavS for GrbS by 4- to 40-fold. The structure reveals how GrbS discriminates VavS specifically from other signaling molecules without binding to the proline-rich motif. PMID- 11406577 TI - Structure of Golgi alpha-mannosidase II: a target for inhibition of growth and metastasis of cancer cells. AB - Golgi alpha-mannosidase II, a key enzyme in N-glycan processing, is a target in the development of anti- cancer therapies. The crystal structure of Drosophila Golgi alpha-mannosidase II in the absence and presence of the anti-cancer agent swainsonine and the inhibitor deoxymannojirimycin reveals a novel protein fold with an active site zinc intricately involved both in the substrate specificity of the enzyme and directly in the catalytic mechanism. Identification of a putative GlcNAc binding pocket in the vicinity of the active site cavity provides a model for the binding of the GlcNAcMan(5)GlcNAc(2) substrate and the consecutive hydrolysis of the alpha1,6- and alpha1,3-linked mannose residues. The enzyme-inhibitor interactions observed provide insight into the catalytic mechanism, opening the door to the design of novel inhibitors of alpha mannosidase II. PMID- 11406578 TI - The B-box dominates SAP-1-SRF interactions in the structure of the ternary complex. AB - The serum response element (SRE) is found in several immediate-early gene promoters. This DNA sequence is necessary and sufficient for rapid transcriptional induction of the human c-fos proto-oncogene in response to stimuli external to the cell. Full activation of the SRE requires the cooperative binding of a ternary complex factor (TCF) and serum response factor (SRF) to their specific DNA sites. The X-ray structure of the human SAP-1-SRF-SRE DNA ternary complex was determined (Protein Data Bank code 1hbx). It shows SAP-1 TCF bound to SRF through interactions between the SAP-1 B-box and SRF MADS domain in addition to contacts between their respective DNA-binding motifs. The SAP-1 B-box is part of a flexible linker of which 21 amino acids become ordered upon ternary complex formation. Comparison with a similar region from the yeast MATalpha2-MCM1 DNA complex suggests a common binding motif through which MADS-box proteins may interact with additional factors such as Fli-1. PMID- 11406579 TI - High-resolution AFM topographs of Rubrivivax gelatinosus light-harvesting complex LH2. AB - Light-harvesting complexes 2 (LH2) are the accessory antenna proteins in the bacterial photosynthetic apparatus and are built up of alphabeta-heterodimers containing three bacteriochlorophylls and one carotenoid each. We have used atomic force microscopy (AFM) to investigate reconstituted LH2 from Rubrivivax gelatinosus, which has a C-terminal hydrophobic extension of 21 amino acids on the alpha-subunit. High-resolution topographs revealed a nonameric organization of the regularly packed cylindrical complexes incorporated into the membrane in both orientations. Native LH2 showed one surface which protruded by approximately 6 A and one that protruded by approximately 14 A from the membrane. Topographs of samples reconstituted with thermolysin-digested LH2 revealed a height reduction of the strongly protruding surface to approximately 9 A, and a change of its surface appearance. These results suggested that the alpha-subunit of R.gelatinosus comprises a single transmembrane helix and an extrinsic C-terminus, and allowed the periplasmic surface to be assigned. Occasionally, large rings ( approximately 120 A diameter) surrounded by LH2 rings were observed. Their diameter and appearance suggest the large rings to be LH1 complexes. PMID- 11406580 TI - Molecular markers of serine protease evolution. AB - The evolutionary history of serine proteases can be accounted for by highly conserved amino acids that form crucial structural and chemical elements of the catalytic apparatus. These residues display non- random dichotomies in either amino acid choice or serine codon usage and serve as discrete markers for tracking changes in the active site environment and supporting structures. These markers categorize serine proteases of the chymotrypsin-like, subtilisin-like and alpha/beta-hydrolase fold clans according to phylogenetic lineages, and indicate the relative ages and order of appearance of those lineages. A common theme among these three unrelated clans of serine proteases is the development or maintenance of a catalytic tetrad, the fourth member of which is a Ser or Cys whose side chain helps stabilize other residues of the standard catalytic triad. A genetic mechanism for mutation of conserved markers, domain duplication followed by gene splitting, is suggested by analysis of evolutionary markers from newly sequenced genes with multiple protease domains. PMID- 11406581 TI - EndoS, a novel secreted protein from Streptococcus pyogenes with endoglycosidase activity on human IgG. AB - Streptococcus pyogenes is an important human pathogen that selectively interacts with proteins involved in the humoral defense system, such as immunoglobulins and complement factors. In this report we show that S.pyogenes has the ability to hydrolyze the chitobiose core of the asparagine-linked glycan on immuno globulin G (IgG) when bacteria are grown in the presence of human plasma. This activity is associated with the secretion of a novel 108 kDa protein denoted EndoS. EndoS has endoglycosidase activity on purified soluble IgG as well as IgG bound to the bacterial surface. EndoS is required for the activity on IgG, as an isogenic EndoS mutant could not hydrolyze the glycan on IgG. In addition, we show that the secreted streptococcal cysteine proteinase SpeB cleaves IgG in the hinge region in a papain-like manner. This is the first example of an endoglycosidase produced by a bacterial pathogen that selectively hydrolyzes human IgG, and reveals a novel mechanism which may contribute to S.pyogenes pathogenesis. PMID- 11406582 TI - The lactose transport protein is a cooperative dimer with two sugar translocation pathways. AB - The Major Facilitator Superfamily lactose transport protein (LacS) undergoes reversible self-association in the detergent-solubilized state, and is present in the membrane as a dimer. We determined the functional unit for proton motive force (Deltap)-driven lactose uptake and lactose/methyl-beta-D-galactopyranoside equilibrium exchange in a proteoliposomal system in which a single cysteine mutant, LacS-C67, defective in Deltap-driven uptake, was co-reconstituted with fully functional cysteine-less protein, LacS-cl. From the quadratic relationship between the uptake activity and the ratio of LacS-C67/LacS-cl, we conclude that the dimeric state of LacS is required for Deltap-driven uptake. N-ethylmaleimide (NEM) treatment of proteoliposomes abolished the LacS-C67 exchange activity but left the LacS-cl unaffected. After NEM treatment, the exchange activity decreased linearly with increasing ratios of LacS-C67/LacS-cl, suggesting that the monomeric state of LacS is sufficient for this mode of transport. We propose that the two subunits of LacS are functionally coupled in the step associated with conformational reorientation of the empty binding site, a step unique for Deltap driven uptake. PMID- 11406583 TI - A switch in disulfide linkage during minicollagen assembly in Hydra nematocysts. AB - The smallest known collagens with only 14 Gly-X-Y repeats referred to as minicollagens are the main constituents of the capsule wall of nematocysts. These are explosive organelles found in Hydra, jellyfish, corals and other Cnidaria. Minicollagen-1 of Hydra recombinantly expressed in mammalian 293 cells contains disulfide bonds within its N- and C-terminal Cys-rich domains but no interchain cross-links. It is soluble and self-associates through non-covalent interactions to form 25-nm-long trimeric helical rod-like molecules. We have used a polyclonal antibody prepared against the recombinant protein to follow the maturation of minicollagens from soluble precursors present in the endoplasmic reticulum and post-Golgi vacuoles to the disulfide-linked insoluble assembly form of the wall. The switch from intra- to intermolecular disulfide bonds is associated with 'hardening' of the capsule wall and provides an explanation for its high tensile strength and elasticity. The process is comparable to disulfide reshuffling between the NC1 domains of collagen IV in mammalian basement membranes. PMID- 11406584 TI - Coordinate gene regulation by fimbriae-induced signal transduction. AB - Fimbriae are thread-like polymers displayed in large amounts on the bacterial surface and used by many pathogens to attach to receptors on host tissue surfaces. Fimbriae contain disulfide bridges, contrary to many Escherichia coli surface proteins produced in bulk amounts. Here we investigate whether fimbriae expression can affect expression of other genes. Analysis of gene expression in two E.coli strains, differing in the fim locus, indicated the flu gene to be affected. The flu gene encodes the antigen 43 (Ag43) surface protein, specifically involved in bacterial aggregation, and microcolony and biofilm formation. Ag43 production is repressed by the global regulator OxyR, which monitors the cell's thiol-disulfide status. Only the thiol form of OxyR represses Ag43 production. We demonstrate that production of several different disulfide containing fimbriae results in the abolition of Ag43 production. No effect was observed in an oxyR mutant. We conclude that fimbriae expression per se constitutes a signal transduction mechanism that affects a number of unrelated genes via the thiol-disulfide status of OxyR. Thus, phase variation in fimbrial expression is coordinated with the expression of other disease- and colonization related genes. PMID- 11406585 TI - The KDEL receptor mediates a retrieval mechanism that contributes to quality control at the endoplasmic reticulum. AB - Newly synthesized proteins in the endoplasmic reticulum (ER) must fold and assemble correctly before being transported to their final cellular destination. While some misfolded or partially assembled proteins have been shown to exit the ER, they fail to escape the early secretory system entirely, because they are retrieved from post-ER compartments to the ER. We elucidate a mechanistic basis for this retrieval and characterize its contribution to ER quality control by studying the fate of the unassembled T-cell antigen receptor (TCR) alpha chain. While the steady-state distribution of TCRalpha is in the ER, inhibition of retrograde transport by COPI induces the accumulation of TCRalpha in post-ER compartments, suggesting that TCRalpha is cycling between the ER and post-ER compartments. TCRalpha associates with BiP, a KDEL protein. Disruption of the ligand-binding function of the KDEL receptor releases TCRalpha from the early secretory system to the cell surface, so that TCRalpha is no longer subject to ER degradation. Thus, our findings suggest that retrieval by the KDEL receptor contributes to mechanisms by which the ER monitors newly synthesized proteins for their proper disposal. PMID- 11406586 TI - Effects of protein stability and structure on substrate processing by the ClpXP unfolding and degradation machine. AB - ClpXP is an ATP-dependent protease that denatures native proteins and translocates the denatured polypeptide into an interior peptidase chamber for degradation. To address the mechanism of these processes, Arc repressor variants with dramatically different stabilities and unfolding half-lives varying from months to seconds were targeted to ClpXP by addition of the ssrA degradation tag. Remarkably, ClpXP degraded each variant at a very similar rate and hydrolyzed approximately 150 molecules of ATP for each molecule of substrate degraded. The hyperstable substrates did, however, slow the ClpXP ATPase cycle. These results confirm that ClpXP uses an active mechanism to denature its substrates, probably one that applies mechanical force to the native structure. Furthermore, the data suggest that denaturation is inherently inefficient or that significant levels of ATP hydrolysis are required for other reaction steps. ClpXP degraded disulfide cross-linked dimers efficiently, even when just one subunit contained an ssrA tag. This result indicates that the pore through which denatured proteins enter the proteolytic chamber must be large enough to accommodate simultaneous passage of two or three polypeptide chains. PMID- 11406587 TI - Direct cleavage of the human DNA fragmentation factor-45 by granzyme B induces caspase-activated DNase release and DNA fragmentation. AB - The protease granzyme B (GrB) plays a key role in the cytocidal activity during cytotoxic T lymphocyte (CTL)-mediated programmed cell death. Multiple caspases have been identified as direct substrates for GrB, suggesting that the activation of caspases constitutes an important event during CTL-induced cell death. However, recent studies have provided evidence for caspase-independent pathway(s) during CTL-mediated apoptosis. In this study, we demonstrate caspase-independent and direct cleavage of the 45 kDa unit of DNA fragmentation factor (DFF45) by GrB both in vitro and in vivo. Using a novel and selective caspase-3 inhibitor, we show the ability of GrB to process DFF45 directly and mediate DNA fragmentation in the absence of caspase-3 activity. Furthermore, studies with DFF45 mutants reveal that both caspase-3 and GrB share a common cleavage site, which is necessary and sufficient to induce DNA fragmentation in target cells during apoptosis. Together, our data suggest that CTLs possess alternative mechanism(s) for inducing DNA fragmentation without the requirement for caspases. PMID- 11406588 TI - Direct inhibition of caspase 3 is dispensable for the anti-apoptotic activity of XIAP. AB - XIAP is a mammalian inhibitor of apoptosis protein (IAP). To determine residues within the second baculoviral IAP repeat (BIR2) required for inhibition of caspase 3, we screened a library of BIR2 mutants for loss of the ability to inhibit caspase 3 toxicity in the yeast Schizosaccharomyces pombe. Four of the mutations, not predicted to affect the structure of the BIR fold, clustered together on the N-terminal region that flanks BIR2, suggesting that this is a site of interaction with caspase 3. Introduction of these mutations into full length XIAP reduced caspase 3 inhibitory activity up to 500-fold, but did not affect its ability to inhibit caspase 9 or interact with the IAP antagonist DIABLO. Furthermore, these mutants retained full ability to inhibit apoptosis in transfected cells, demonstrating that although XIAP is able to inhibit caspase 3, this activity is dispensable for inhibition of apoptosis by XIAP in vivo. PMID- 11406589 TI - Sec61p-independent degradation of the tail-anchored ER membrane protein Ubc6p. AB - Tail-anchored proteins are distinct from other membrane proteins as they are thought to insert into the endoplasmic reticulum (ER) membrane independently of Sec61p translocation pores. These pores not only mediate import but are also assumed to catalyze export of proteins in a process called ER-associated protein degradation (ERAD). In order to examine the Sec61p dependence of the export of tail-anchored proteins, we analyzed the degradation pathway of a tail-anchored ER membrane protein, the ubiquitin-conjugating enzyme 6 (Ubc6p). In contrast to other ubiquitin conjugating enzymes (Ubcs), Ubc6p is naturally short-lived. Its proteolysis is mediated specifically by the unique Ubc6p tail region. Degradation further requires the activity of Cue1p-assembled Ubc7p, and its own catalytic site cysteine. However, it occurs independently of the other ERAD components Ubc1p, Hrd1p/Der3p, Hrd3p and Der1p. In contrast to other natural ERAD substrates, proteasomal mutants accumulate a membrane-bound degradation intermediate of Ubc6p. Most interestingly, mutations in SEC61 do not reduce the turnover of full-length Ubc6p nor cause a detectable accumulation of degradation intermediates. These data are in accordance with a model in which tail-anchored proteins can be extracted from membranes independently of Sec61p. PMID- 11406590 TI - Toxoplasma evacuoles: a two-step process of secretion and fusion forms the parasitophorous vacuole. AB - Rapid discharge of secretory organelles called rhoptries is tightly coupled with host cell entry by the protozoan parasite Toxoplasma gondii. Rhoptry contents were deposited in clusters of vesicles within the host cell cytosol and within the parasitophorous vacuole. To examine the fate of these rhoptry-derived secretory vesicles, we utilized cytochalasin D to prevent invasion, leading to accumulation of protein-rich vesicles in the host cell cytosol. These vesicles lack an internal parasite and are hence termed evacuoles. Like the mature parasite-containing vacuole, evacuoles became intimately associated with host cell mitochondria and endoplasmic reticulum, while remaining completely resistant to fusion with host cell endosomes and lysosomes. In contrast, evacuoles were recruited to pre-existing, parasite-containing vacuoles and were capable of fusing and delivering their contents to these compartments. Our findings indicate that a two-step process involving direct rhoptry secretion into the host cell cytoplasm followed by incorporation into the vacuole generates the parasitophorous vacuole occupied by TOXOPLASMA: The characteristic properties of the mature vacuole are likely to be determined by this early delivery of rhoptry components. PMID- 11406591 TI - Vac8p release from the SNARE complex and its palmitoylation are coupled and essential for vacuole fusion. AB - Activated fatty acids stimulate budding and fusion in several cell-free assays for vesicular transport. This stimulation is thought to be due to protein palmitoylation, but relevant substrates have not yet been identified. We now report that Vac8p, a protein known to be required for vacuole inheritance, becomes palmitoylated when isolated yeast vacuoles are incubated under conditions that allow membrane fusion. Similar requirements for Vac8p palmitoylation and vacuole fusion, the inhibition of vacuole fusion by antibodies to Vac8p and the strongly reduced fusion of vacuoles lacking Vac8p suggest that palmitoylated Vac8p is essential for homotypic vacuole fusion. Strikingly, palmitoylation of Vac8p is blocked by the addition of antibodies to Sec18p (yeast NSF) only. Consistent with this, a portion of Vac8p is associated with the SNARE complex on vacuoles, which is lost during Sec18p- and ATP-dependent priming. During or after SNARE complex disassembly, palmitoylation occurs and anchors Vac8p to the vacuolar membrane. We propose that palmitoylation of Vac8p is regulated by the same machinery that controls membrane fusion. PMID- 11406592 TI - The distal region and receptor tyrosines of the Epo receptor are non-essential for in vivo erythropoiesis. AB - The erythropoietin receptor (EpoR) is required for the proliferation and survival of committed erythroid lineage cells. Previous studies have utilized receptor mutations to show the requirement for the distal half of the cytoplasmic domain of the EpoR and receptor tyrosines for activation of signaling pathways potentially critical to Epo function. To extend these studies to in vivo erythropoiesis, we have created two mutant strains of mice. One strain (H) contains a truncation of the distal half of the cytoplasmic domain, while the second strain (HM) contains the same truncation as well as the mutation of the residual tyrosine (Y(343)) to a phenylalanine. Strikingly, both strains of mice are viable, with only slight alterations in constitutive erythropoiesis or in in vitro assays of red cell lineage function. Challenging H mutant mice with continuous injections of Epo results in an erythrocytosis that is not seen in HM mice. The results demonstrate that neither the distal region nor receptor tyrosines are essential for in vivo EpoR function, but contribute to receptor function in a subtle manner. PMID- 11406593 TI - Cell signaling can direct either binary or graded transcriptional responses. AB - Transcriptional control is generally thought to operate as a binary switch, a behavior that might explain observations such as monoallelic gene expression, stochastic phenotypic changes and bimodal gene activation kinetics. By measuring the activity of the single-copy GAL1 promoter in single cells, we found that changes in the activities of either the transcriptional activator, Gal4 (by simple recruitment with synthetic ligands), or the transcriptional repressor, Mig1, generated graded (non-binary) changes in gene expression that were proportional to signal intensity. However, in the context of the endogenous glucose-responsive signaling pathway, these transcription factors formed part of a binary transcriptional response. Genetic studies demonstrated that this binary response resulted from regulation of a second repressor, Gal80, whereas regulation of Mig1 by a distinct signaling pathway generated graded changes in GAL1 promoter activity. Surprisingly, isogenetic cells can respond to glucose with either binary or graded changes in gene expression, depending on growth conditions. Our studies demonstrate that a given promoter can adapt either binary or graded behavior, and identify the Mig1 and Gal80 genes as necessary for binary versus graded behavior of the Gal1 promoter. PMID- 11406594 TI - Transcript analysis of 1003 novel yeast genes using high-throughput northern hybridizations. AB - The expression of 1008 open reading frames (ORFs) from the yeast Saccharomyces cerevisiae has been examined under eight different physiological conditions, using classical northern analysis. These northern data have been compared with publicly available data from a microarray analysis of the diauxic transition in S.cerevisiae. The results demonstrate the importance of comparing biologically equivalent situations and of the standardization of data normalization procedures. We have also used our northern data to identify co-regulated gene clusters and define the putative target sites of transcriptional activators responsible for their control. Clusters containing genes of known function identify target sites of known activators. In contrast, clusters comprised solely of genes of unknown function usually define novel putative target sites. Finally, we have examined possible global controls on gene expression. It was discovered that ORFs that are highly expressed following a nutritional upshift tend to employ favoured codons, whereas those overexpressed in starvation conditions do not. These results are interpreted in terms of a model in which competition between mRNA molecules for translational capacity selects for codons translated by abundant tRNAs. PMID- 11406595 TI - UV-damaged DNA-binding protein in the TFTC complex links DNA damage recognition to nucleosome acetylation. AB - Initiation of transcription of protein-encoding genes by RNA polymerase II (Pol II) was thought to require transcription factor TFIID, a complex comprised of the TATA box-binding protein (TBP) and TBP-associated factors (TAF(II)s). In the presence of TBP-free TAF(II) complex (TFTC), initiation of Pol II transcription can occur in the absence of TFIID. TFTC containing the GCN5 acetyltransferase acetylates histone H3 in a nucleosomal context. We have identified a 130 kDa subunit of TFTC (SAP130) that shares homology with the large subunit of UV damaged DNA-binding factor. TFTC preferentially binds UV-irradiated DNA, UV damaged DNA inhibits TFTC-mediated Pol II transcription and TFTC is recruited in parallel with the nucleotide excision repair protein XP-A to UV-damaged DNA. TFTC preferentially acetylates histone H3 in nucleosomes assembled on UV-damaged DNA. In agreement with this, strong histone H3 acetylation occurs in intact cells after UV irradiation. These results suggest that the access of DNA repair machinery to lesions within chromatin may be facilitated by TFTC via covalent modification of chromatin. Thus, our experiments reveal a molecular link between DNA damage recognition and chromatin modification. PMID- 11406596 TI - Recognition of polyadenylation sites in yeast pre-mRNAs by cleavage and polyadenylation factor. AB - Recognition of poly(A) sites in yeast pre-mRNAs is poorly understood. Employing an in vitro cleavage system with cleavage and polyadenylation factor (CPF) and cleavage factor IA we show that the efficiency and positioning elements are dispensable for poly(A)-site recognition within a short CYC1 substrate in vitro. Instead, U-rich elements immediately upstream and downstream of the poly(A) site mediate cleavage-site recognition within CYC1 and ADH1 pre-mRNAs. These elements act in concert with the poly(A) site to produce multiple recognition sites for the processing machinery, since combinations of mutations within these elements were most effective in cleavage inhibition. Intriguingly, introduction of a U rich element downstream of the GAL7 poly(A) site strongly enhanced cleavage, underscoring the importance of downstream sequences in general. RNA- binding analyses demonstrate that cleavage depends on the recognition of the poly(A)-site region by CPF. Consistent with in vitro results, mutation of sequences upstream and downstream of the poly(A) site affected 3'-end formation in vivo. A model for yeast pre-mRNA cleavage-site recognition outlines an unanticipated high conservation of yeast and mammalian 3'-end processing mechanisms. PMID- 11406597 TI - The McrBC restriction endonuclease assembles into a ring structure in the presence of G nucleotides. AB - McrBC from Escherichia coli K-12 is a restriction enzyme that belongs to the family of AAA(+) proteins and cuts DNA containing modified cytosines. Two proteins are expressed from the mcrB gene: a full-length version, McrB(L), and a short version, McrB(S). McrB(L) binds specifically to the methylated recognition site and is, therefore, the DNA-binding moiety of the McrBC endonuclease. McrB(S) is devoid of DNA-binding activity. We observed that the quaternary structure of the endonuclease depends on binding of the cofactors. In gel filtration experiments, McrB(L) and McrB(S) form high molecular weight oligomers in the presence of Mg(2+) and GTP, GDP or GTP-gamma-S. Oligomerization did not require the presence of DNA and was independent of GTP hydrolysis. Electron micrographs of negatively stained McrB(L) and McrB(S) revealed ring-shaped particles with a central channel. Mass analysis by scanning transmission electron microscopy indicates that McrB(L) and McrB(S) form single heptameric rings as well as tetradecamers. In the presence of McrC, a subunit that is essential for DNA cleavage, the tetradecameric species was the major form of the endonuclease. PMID- 11406598 TI - Specific targeting of insect and vertebrate telomeres with pyrrole and imidazole polyamides. AB - DNA minor groove-binding compounds (polyamides) that target insect and vertebrate telomeric repeats with high specificity were synthesized. Base pair recognition of these polyamides is based on the presence of the heterocyclic amino acids pyrrole and imidazole. One compound (TH52B) interacts uniquely and with excellent specificity (K(d) = 0.12 nM) with two consecutive insect-type telomeric repeats (TTAGG). A related compound, TH59, displays high specificity (K(d) = 0.5 nM) for tandem vertebrate (TTAGGG) and insect telomeric repeats. The high affinity and specificity of these compounds were achieved by bidentate binding of two flexibly linked DNA-binding moieties. Epifluorescence microscopy studies show that fluorescent derivatives of TH52B and TH59 stain insect or vertebrate telomeres of chromosomes and nuclei sharply. Importantly, the telomere-specific polyamide signals of HeLa chromosomes co-localize with the immunofluorescence signals of the telomere-binding protein TRF1. Our results demonstrate that telomere-specific compounds allow rapid estimation of relative telomere length. The insect-specific compound TH52 was shown to be incorporated rapidly into growing Sf9 cells, underlining the potential of these compounds for telomere biology and possibly human medicine. PMID- 11406599 TI - Telomere resolution in the Lyme disease spirochete. AB - The genus Borrelia includes the causative agents of Lyme disease and relapsing fever. An unusual feature of these bacteria is a genome that includes linear DNA molecules with covalently closed hairpin ends referred to as telomeres. We have investigated the mechanism by which the hairpin telomeres are processed during replication. A synthetic 140 bp sequence having the predicted structure of a replicated telomere was shown to function as a viable substrate for telomere resolution in vivo, and was sufficient to convert a circular replicon to a linear form. Our results suggest that the final step in the replication of linear Borrelia replicons is a site-specific DNA breakage and reunion event to regenerate covalently closed hairpin ends. The telomere substrate described here will be valuable both for in vivo manipulation of linear DNA in Borrelia and for in vitro studies to identify and characterize the telomere resolvase. PMID- 11406600 TI - Bimodal activation of SMC ATPase by intra- and inter-molecular interactions. AB - Structural maintenance of chromosomes (SMC) proteins play fundamental roles in higher-order chromosome dynamics from bacteria to humans. It has been proposed that the Bacillus subtilis SMC (BsSMC) homodimer is composed of two anti-parallel coiled-coil arms, each having an ATP-binding domain at its distal end. It remains totally unknown, however, how the two-armed structure supports ATP-dependent actions of BsSMC. By constructing a number of mutant derivatives including 'single-armed' BsSMC, we show here that the central hinge domain provides a structural flexibility that allows opening and closing of the two arms. This unique structure brings about bimodal regulation of the SMC ATPase cycle. Closing the arm can trigger ATP hydrolysis by allowing an end-end interaction within a dimer (intramolecular mode). When bound to DNA, ATP promotes a dimer-dimer interaction, which in turn activates their DNA-dependent ATPase activity (intermolecular mode). Our results reveal a novel mechanism of ATPase regulation and provide mechanistic insights into how eukaryotic SMC protein complexes could mediate diverse chromosomal functions, such as chromosome condensation and sister chromatid cohesion. PMID- 11406601 TI - Product analysis illuminates the final steps of IES deletion in Tetrahymena thermophila. AB - DNA sequences (IES elements) eliminated from the developing macronucleus in the ciliate Tetrahymena thermophila are released as linear fragments, which have now been detected and isolated. A PCR-mediated examination of fragment end structures reveals three types of strand scission events, reflecting three steps in the deletion process. New evidence is provided for two steps proposed previously: an initiating double-stranded cleavage, and strand transfer to create a branched deletion intermediate. The fragment ends provide evidence for a previously uncharacterized third step: the branched DNA strand is cleaved at one of several defined sites located within 15-16 nucleotides of the IES boundary, liberating the deleted DNA in a linear form. PMID- 11406602 TI - A broad host range replicon with different requirements for replication initiation in three bacterial species. AB - Plasmid RK2 is unusual in its ability to replicate stably in a wide range of Gram negative bacteria. The replication origin (oriV) and a plasmid-encoded initiation protein (TrfA; expressed as 33 and 44 kDa forms) are essential for RK2 replication. To examine initiation events in bacteria unrelated to Escherichia coli, the genes encoding the replicative helicase, DnaB, of Pseudomonas putida and Pseudomonas aeruginosa were isolated and used to construct protein expression vectors. The purified proteins were tested for activity along with E.coli DnaB at RK2 oriV. Each helicase could be recruited and activated at the RK2 origin in the presence of the host-specific DnaA protein and the TrfA protein. Escherichia coli or P.putida DnaB was active with either TrfA-33 or TrfA-44, while P.aeruginosa DnaB required TrfA-44 for activation. Moreover, unlike the E.coli DnaB helicase, both Pseudomonas helicases could be delivered and activated at oriV in the absence of an ATPase accessory protein. Thus, a DnaC-like accessory ATPase is not universally required for loading the essential replicative helicase at a replication origin. PMID- 11406603 TI - Role of the non-homologous DNA end joining pathway in the early steps of retroviral infection. AB - Early after infection, the retroviral RNA genome is reverse transcribed to generate a linear cDNA copy, then that copy is integrated into a chromosome of the host cell. We report that unintegrated viral cDNA is a substrate for the host cell non-homologous DNA end joining (NHEJ) pathway, which normally repairs cellular double-strand breaks by end ligation. NHEJ activity was found to be required for an end-ligation reaction that circularizes a portion of the unintegrated viral cDNA in infected cells. Consistent with this, the NHEJ proteins Ku70 and Ku80 were found to be bound to purified retroviral replication intermediates. Cells defective in NHEJ are known to undergo apoptosis in response to retroviral infection, a response that we show requires reverse transcription to form the cDNA genome but not subsequent integration. We propose that the double-strand ends present in unintegrated cDNA promote apoptosis, as is known to be the case for chromosomal double-strand breaks, and cDNA circularization removes the pro-apoptotic signal. PMID- 11406604 TI - DNA helicase-mediated packaging of adeno-associated virus type 2 genomes into preformed capsids. AB - Helicases not only catalyse the disruption of hydrogen bonding between complementary regions of nucleic acids, but also move along nucleic acid strands in a polar fashion. Here we show that the Rep52 and Rep40 proteins of adeno associated virus type 2 (AAV-2) are required to translocate capsid-associated, single-stranded DNA genomes into preformed empty AAV-2 capsids, and that the DNA helicase function of Rep52/40 is essential for this process. Furthermore, DNase protection experiments suggest that insertion of AAV-2 genomes proceeds from the 3' end, which correlates with the 3'-->5' processivity demonstrated for the Rep52/40 helicase. A model is proposed in which capsid-immobilized helicase complexes act as molecular motors to 'pump' single-stranded DNA across the capsid boundary. PMID- 11406605 TI - Positional dissociation between the genetic mutation responsible for pseudohypoparathyroidism type Ib and the associated methylation defect at exon A/B: evidence for a long-range regulatory element within the imprinted GNAS1 locus. AB - Pseudohypoparathyroidism type Ib (PHP-Ib) is a paternally imprinted disorder which maps to a region on chromosome 20q13.3 that comprises GNAS1 at its telomeric boundary. Exon A/B of this gene was recently shown to display a loss of methylation in several PHP-Ib patients. In nine unrelated PHP-Ib kindreds, in whom haplotype analysis and mode of inheritance provided no evidence against linkage to this chromosomal region, we confirmed lack of exon A/B methylation for affected individuals, while unaffected carriers showed no epigenetic abnormality at this locus. However, affected individuals in one kindred (Y2) displayed additional methylation defects involving exons NESP55, AS and XL, and unaffected carriers in this family showed an abnormal methylation at exon NESP55, but not at other exons. Taken together, current evidence thus suggests that distinct mutations within or close to GNAS1 can lead to PHP-Ib and the associated epigenetic changes. To further delineate the telomeric boundary of the PHP-Ib locus, the previously reported kindred F, in which patient F-V/51 is recombinant within GNAS1, was investigated with several new markers and direct nucleotide sequence analysis. These studies revealed that F-V/51 remains recombinant at a single nucleotide polymorphism (SNP) located 1.2 kb upstream of XL. No heterozygous mutation was identified between exon XL and an SNP approximately 8 kb upstream of NESP55, where this affected individual becomes linked, suggesting that the genetic defect responsible for parathyroid hormone resistance in kindred F, and probably other PHP-Ib patients, is located >or=56 kb centromeric of the abnormally methylated exon A/B. A region upstream of the known coding exons of GNAS1 is therefore predicted to exert, presumably through imprinting of exon A/B, long-range effects on G(s)alpha expression. PMID- 11406606 TI - Expanded CAG repeats in exon 1 of the Huntington's disease gene stimulate dopamine-mediated striatal neuron autophagy and degeneration. AB - Huntington's disease (HD) is caused by an expanded CAG repeat in exon 1 of the gene coding for the huntingtin protein. The cellular pathway by which this mutation induces HD remains unknown, although alterations in protein degradation are involved. To study intrinsic cellular mechanisms linked to the mutation, we examined dissociated postnatally derived cultures of striatal neurons from transgenic mice expressing exon 1 of the human HD gene carrying a CAG repeat expansion. While there was no difference in cell death between wild-type and mutant littermate-derived cultures, the mutant striatal neurons exhibited elevated cell death following a single exposure to a neurotoxic concentration of dopamine. The mutant neurons exposed to dopamine also exhibited lysosome associated responses including induction of autophagic granules and electron dense lysosomes. The autophagic/lysosomal compartments co-localized with high levels of oxygen radicals in living neurons, and ubiquitin. The results suggest that the combination of mutant huntingtin and a source of oxyradical stress (provided in this case by dopamine) induces autophagy and may underlie the selective cell death characteristic of HD. PMID- 11406607 TI - Highly activated Fgfr3 with the K644M mutation causes prolonged survival in severe dwarf mice. AB - Several gain-of-function mutations in a receptor tyrosine kinase, fibroblast growth factor receptor 3 (FGFR3), cause dwarfism in humans. Two particularly severe dwarfisms, thanatophoric dysplasia type II (TDII) and severe achondroplasia with developmental delay and acanthosis nigricans (SADDAN), are associated with glutamic acid (E) and methionine (M) substitutions at the K650 residue in the kinase domain. TDII is lethal at birth, whereas most of the SADDAN patients survive the perinatal period. However, FGFR3 with the SADDAN mutation is more activated than FGFR3 with the TDII mutation in vitro. To find out whether the K650M mutation also causes the SADDAN phenotype, we introduced the corresponding point mutation (K644M) into the mouse Fgfr3 gene. Heterozygous mutant mice show a phenotype similar to human SADDAN, e.g. the majority of the SADDAN mice survive the perinatal period. This suggests that the survival of SADDAN patients is indeed attributed to the K650M mutation in FGFR3. The long bone abnormalities in SADDAN mice are milder than the TDII model. In addition, overgrowth of the cartilaginous tissues is observed in the rib cartilage, trachea and nasal septum. The FGF ligand at the low concentration differentially activates Map kinase in primary chondrocyte cultures from wild-type and SADDAN mice. Comparisons of the molecular bases of the phenotypic differences in SADDAN and TDII mice may increase our understanding of the factors that influence the severity in these two related skeletal dysplasias. PMID- 11406608 TI - Genetic polymorphisms of biotransformation enzymes in patients with Hodgkin's and non-Hodgkin's lymphomas. AB - Considering the role in the metabolism of chemicals played by biotransformation enzymes, we aimed at determining whether any association exists between genetic polymorphisms in CYP1A1, CYP2E1, epoxide hydrolase (EPHX), glutathione S transferases (GSTM1/P1/T1) and individual susceptibility to lymphomas. PCR-RFLP based genotyping assays were used to determine the frequency of polymorphisms in CYP1A1 (3'-flanking region), CYP2E1 (5'-flanking region and intron 6), EPHX (exons 3 and 4), GSTM1 (deletion), GSTP1 (exon 5) and GSTT1 (deletion) in a case control study comprised of 219 patients with morbus Hodgkin (MH) and non Hodgkin's lymphomas (NHL) and 455 age- and sex-matched healthy individuals. The distribution of genotypes in CYP2E1-intron 6 was significantly different between the control group and all lymphomas (P = 0.03), patients with NHL (P = 0.024), and especially aggressive diffuse NHL (P = 0.007). Grading of NHL seemed to be associated with this polymorphism as well (P = 0.041). The EPHX-exon 3 genotype distribution was significantly different between control males and males with all lymphomas (P = 0.01) or with NHL (P = 0.019). The Val/Val genotype of GSTP1-exon 5 was prevalent in all MH [odds ratio (OR) = 2.08, 95% confidence interval (CI) = 1.05-4.14] and this difference was particularly evident in females (OR = 2.97, 95% CI = 1.16-7.61). A significant difference in the distribution of GSTP1-exon 5 genotypes was found between NHL tumors >5 cm and those <5 cm (P = 0.03). The results suggest that genetic polymorphisms of biotransformation enzymes may play a significant role in the development of lymphoid malignancies. PMID- 11406609 TI - B-cell neoplasia associated gene with multiple splicing (BCMS): the candidate B CLL gene on 13q14 comprises more than 560 kb covering all critical regions. AB - Deletions in chromosomal band 13q14.3 occur in >50% of B-cell chronic lymphocytic leukemias (B-CLL) and mantle cell lymphoma, indicating the localization of a tumor suppressor gene involved in the pathomechanism of these diseases. Within a 400 kb recurrently deleted segment at least two minimally deleted subregions had been reported. For the two genes residing in the proximal subregion, initially named LEU1 and LEU2, a pathogenic role has not yet been established. We report here that LEU1 is only a small portion of a large gene, which spans all previously reported critical subregions including the distal subregion. This gene, designated B-cell neoplasia-associated gene with multiple splicing (BCMS), is composed of at least 50 exons spanning >or=560 kb of genomic DNA and is expressed in more than 20 RNA splicing variants. While tissue-specific expression of RNA variants was observed, there was no evidence for the expression of a variant specific for B-CLL. Sequence analysis of the RNA variants suggests that BCMS transcripts belong to the group of non-coding RNAs. The alignment of the gene with all critical subregions provides a strong argument for BCMS being the most likely candidate for the tumor suppressor gene in 13q14 involved in the leukemogenesis of B-CLL. Due to the limited understanding of functional RNAs, however, it remains difficult to prove the pathogenic role of BCMS. PMID- 11406610 TI - Evidence for BLM and Topoisomerase IIIalpha interaction in genomic stability. AB - The genomic instability of persons with Bloom's syndrome (BS) features particularly an increased number of sister-chromatid exchanges (SCEs). The primary cause of the genomic instability is mutation at BLM, which encodes a DNA helicase of the RecQ family. BLM interacts with Topoisomerase IIIalpha (Topo IIIalpha), and both BLM and Topo IIIalpha localize to the nuclear organelles referred to as the promyelocytic leukemia protein (PML) nuclear bodies. In this study we show, by analysis of cells that express various deletion constructs of green fluorescent protein (GFP)-tagged BLM, that the first 133 amino acids of BLM are necessary and sufficient for interaction between Topo IIIalpha and BLM. The Topo IIIalpha-interaction domain of BLM is not required for BLM's localization to the PML nuclear bodies; in contrast, Topo IIIalpha is recruited to the PML nuclear bodies via its interaction with BLM. Expression of a full-length BLM (amino acids 1-1417) in BS cells can correct their high SCEs to normal levels, whereas expression of a BLM fragment that lacks the Topo IIIalpha interaction domain (amino acids 133-1417) results in intermediate SCE levels. The deficiency of amino acids 133-1417 in the reduction of SCEs was not explained by a defect in DNA helicase activity, because immunoprecipitated 133-1417 protein had 4-fold higher activity than GFP-BLM. The data implicate the BLM-Topo IIIalpha complex in the regulation of recombination in somatic cells. PMID- 11406611 TI - Cloning of the human MCCA and MCCB genes and mutations therein reveal the molecular cause of 3-methylcrotonyl-CoA: carboxylase deficiency. AB - 3-Methylcrotonyl-CoA: carboxylase (EC 6.4.1.4; MCC) deficiency is an inborn error of the leucine degradation pathway (MIM *210200) characterized by increased urinary excretion of 3-hydroxyisovaleric acid and 3-methylcrotonylglycine. The clinical phenotypes are highly variable ranging from asymptomatic to profound metabolic acidosis and death in infancy. Sequence similarity with Glycine max and Arabidopsis thaliana genes encoding the two subunits of MCC permitted us to clone the cDNAs encoding the alpha- and beta-subunits of human MCC. The 2580 bp MCCA cDNA encodes the 725 amino acid biotin-containing alpha-subunit. The MCCA gene is located on chromosome 3q26-q28 and consists of 19 exons. The 2304 bp MCCB cDNA encodes the non-biotin-containing beta-subunit of 563 amino acids. The MCCB gene is located on chromosome 5q13 and consists of 17 exons. We have sequenced both genes in four patients with isolated biotin-unresponsive deficiency of MCC. In two of them we found mutations in the MCCA gene. Compound heterozygosity for a missense mutation (S535F) and a nonsense mutation (V694X) were identified in one patient. One heterozygous mutation (S535F) was found in another patient. The remaining two patients had mutations in the MCCB gene. One consanguineous patient was homozygous for a missense mutation (R268T). In the other we identified a missense mutation in one allele (E99Q) and allelic loss of the other. Mutations were correlated with an almost total lack of enzyme activity in fibroblasts. These data provide evidence that human MCC deficiency is caused by mutations in either the MCCA or MCCB gene. PMID- 11406612 TI - Geldanamycin activates a heat shock response and inhibits huntingtin aggregation in a cell culture model of Huntington's disease. AB - Huntington's disease (HD) is a progressive neurodegenerative disorder with no effective treatment. Geldanamycin is a benzoquinone ansamycin that binds to the heat shock protein Hsp90 and activates a heat shock response in mammalian cells. In this study, we show by using a filter retardation assay and immunofluorescence microscopy that treatment of mammalian cells with geldanamycin at nanomolar concentrations induces the expression of Hsp40, Hsp70 and Hsp90 and inhibits HD exon 1 protein aggregation in a dose-dependent manner. Similar results were obtained by overexpression of Hsp70 and Hsp40 in a separate cell culture model of HD. This is the first demonstration that huntingtin protein aggregation in cells can be suppressed by chemical compounds activating a specific heat shock response. These findings may provide the basis for the development of a novel pharmacotherapy for HD and related glutamine repeat disorders. PMID- 11406613 TI - BACE knockout mice are healthy despite lacking the primary beta-secretase activity in brain: implications for Alzheimer's disease therapeutics. AB - Alzheimer's disease (AD) is a neurodegenerative disorder characterized by accumulation of amyloid plaques and neurofibrillary tangles in the brain. The major components of plaque, beta-amyloid peptides (Abetas), are produced from amyloid precursor protein (APP) by the activity of beta- and gamma-secretases. beta-secretase activity cleaves APP to define the N-terminus of the Abeta1-x peptides and, therefore, has been a long- sought therapeutic target for treatment of AD. The gene encoding a beta-secretase for beta-site APP cleaving enzyme (BACE) was identified recently. However, it was not known whether BACE was the primary beta-secretase in mammalian brain nor whether inhibition of beta secretase might have effects in mammals that would preclude its utility as a therapeutic target. In the work described herein, we generated two lines of BACE knockout mice and characterized them for pathology, beta-secretase activity and Abeta production. These mice appeared to develop normally and showed no consistent phenotypic differences from their wild-type littermates, including overall normal tissue morphology and brain histochemistry, normal blood and urine chemistries, normal blood-cell composition, and no overt behavioral and neuromuscular effects. Brain and primary cortical cultures from BACE knockout mice showed no detectable beta-secretase activity, and primary cortical cultures from BACE knockout mice produced much less Abeta from APP. The findings that BACE is the primary beta-secretase activity in brain and that loss of beta-secretase activity produces no profound phenotypic defects with a concomitant reduction in beta-amyloid peptide clearly indicate that BACE is an excellent therapeutic target for treatment of AD. PMID- 11406614 TI - Neural tube defects and neuroepithelial cell death in Tulp3 knockout mice. AB - The tubby-like protein 3 (Tulp3) gene has been identified as a member of a small novel gene family which is primarily neuronally expressed. Mutations in two of the family members, tub and tulp1, have been shown to cause neurosensory disorders. To determine the in vivo function of Tulp3, we have generated a germline mutation in the mouse Tulp3 gene by homologous recombination. Embryos homozygous for the Tulp3 mutant allele exhibit failure of neural tube closure, and die by embryonic day 14.5. Failure of cranial neural tube closure coincided with increased neuroepithelial apoptosis specifically in the hindbrain and the caudal neural tube. In addition, the number of betaIII-tubulin positive cells is significantly decreased in the hindbrain of Tulp3(-/-) embryos. These results suggest that disruption of the Tulp3 gene affects the development of a neuronal cell population. Interestingly, some Tulp3 heterozygotes also manifest embryonic lethality with neuroepithelial cell death. Our results demonstrate that the Tulp3 gene is essential for embryonic development in mice. PMID- 11406615 TI - The brain exocyst complex interacts with RalA in a GTP-dependent manner: identification of a novel mammalian Sec3 gene and a second Sec15 gene. AB - Ral is a small GTPase involved in critical cellular signaling pathways. The two isoforms, RalA and RalB, are widely distributed in different tissues, with RalA being enriched in brain. The best characterized RalA signaling pathways involve RalBP1 and phospholipase D. To investigate RalA signaling in neuronal cells we searched for RalA-binding proteins in brain. We found at least eight proteins that bound RalA in a GTP-dependent manner. Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) identified these as the components of the exocyst complex. The yeast exocyst is a regulator of polarized secretion, docking vesicles to regions of the plasma membrane involved in active exocytosis. We identified the human FLJ10893 protein as the mammalian homologue of the yeast exocyst protein Sec3p. The exocyst complex did not contain the previously identified exocyst component rSec15, but a new homologue of both yeast Sec15p and rSec15, called KIAA0919. Western blots confirmed that two rat exocyst proteins, rSec6 and rSec8, bound active RalA in nerve terminals, as did RalBP1. Phospholipase D bound RalA in a nucleotide-independent manner. This places the RalA signaling system in mammalian nerve terminals, where the exocyst may act as an effector for activated RalA in directing sites of exocytosis. PMID- 11406616 TI - p21Cip-1/SDI-1/WAF-1 gene is involved in chondrogenic differentiation of ATDC5 cells in vitro. AB - Development of skeletal cartilage is characterized with coupling growth arrest and cell differentiation. Here, to understand the cyclin-dependent kinase inhibitors involved in the progression of chondrogenic differentiation, we examined changes in the expression levels of cyclin-dependent kinase inhibitor members using mouse ATDC5 prechondrocytes as a widely used in vitro model of cartilage differentiation. Up-regulation of p21 and p27 mRNA was observed following a decrease in growth rate of prechondrocytes, and both transcripts subsequently accumulated during chondrogenic differentiation; p15, p18, and p19 mRNA, in contrast, did not change during differentiation. Only the up-regulation of p21 mRNA during differentiation was prevented by the continuous treatment of early chondrogenic inhibitor, parathyroid hormone, indicating a close correlation between differentiation and p21 induction in ATDC5 cells. Therefore, to examine the role of p21 during chondrogenesis, we established stable cell lines overexpressing full-length p21 antisense RNA in ATDC5. The reduction of endogenous p21 in these cell lines caused inhibition of early chondrogenic differentiation in ATDC5, indicating that p21 gene plays an important role in this process of the cells in vitro. Furthermore, the level of p21 protein and p21.CDK2 complexes transiently increased during differentiation, but not in undifferentiated cells, leading to a decrease in CDK2-associated kinase. However, differentiation-dependent expressed p21 protein was degraded by a proteasome dependent pathway. Thus, the progression of chondrogenic differentiation requires down-regulation of CDK2-associated kinase with an increase in p21 protein and subsequent degradation of this protein by a proteasomal pathway. PMID- 11406617 TI - A composite element binding the vitamin D receptor and the retinoic X receptor alpha mediates the transforming growth factor-beta inhibition of decorin gene expression in articular chondrocytes. AB - Decorin, a small leucine-rich proteoglycan may play an important role in the attempt of cartilage repair initiated by chondrocytes in early stages of osteoarthritis, through its ability to bind collagen fibrils and growth factors such as transforming growth factor-beta (TGF-beta). We previously demonstrated that TGF-beta decreased decorin mRNA steady state levels in articular chondrocytes (Demoor, M., Redini, F., Boittin, M., and Pujol, J.-P. (1998) Biochim. Biophys. Acta 1398, 179-191). Here, we investigated the effect of TGF beta on decorin gene expression in both primary cultures of articular chondrocytes and chondrocytes dedifferentiated by serial passages. Transient transfection of cells with plasmid constructs of the decorin promoter linked to the luciferase reporter gene revealed transcriptional repression by TGF-beta, in fully differentiated as well as dedifferentiated chondrocytes. Experiments with 5'-deleted constructs allowed characterization of a TGF-beta-responsive element in the shortest construct (base pairs (bp) -155/+269). DNase I footprinting analysis delineated a negative TGF-beta-responsive region between -140 and -111 bp in the decorin proximal promoter. Gel retardation assays demonstrated that TGF beta modulates decorin gene expression through transcription factors, the nature and mode of action of which depend on the differentiation state of the chondrocytes; two DNA-protein complexes were formed in the region -144/-127 bp with nuclear extracts from primary chondrocytes, whereas a higher mobility complex was observed in the -127/-111 bp region for dedifferentiated cells. Antibodies against vitamin D and retinoic acid receptors used in supershift experiments showed that these nuclear receptors are involved in the regulation of decorin gene expression in articular chondrocytes. PMID- 11406618 TI - Neurons are protected from excitotoxic death by p53 antisense oligonucleotides delivered in anionic liposomes. AB - The potential of anionic liposomes for oligonucleotide delivery was explored because the requirement for a net-positive charge on transfection-competent cationic liposome-DNA complexes is ambiguous. Liposomes composed of phosphatidylglycerol and phosphatidylcholine were monodisperse and encapsulated oligonucleotides with 40-60% efficiency. Ionic strength, bilayer charge density, and oligonucleotide chemistry influenced encapsulation. To demonstrate the biological efficacy of this vector, antisense oligonucleotides to p53 delivered in anionic liposomes were tested in an in vitro model of excitotoxicity. Exposure of hippocampal neurons to glutamate increased p53 protein expression 4-fold and decreased neuronal survival to approximately 35%. Treatment with 1 microm p53 antisense oligonucleotides in anionic liposomes prevented glutamate-induced up regulation of p53 and increased neuronal survival to approximately 75%. Encapsulated phosphorothioate p53 antisense oligonucleotides were neuroprotective at 5-10-fold lower concentrations than when unencapsulated. Replacing the anionic lipid with phosphatidylserine significantly decreased neuroprotection. p53 antisense oligonucleotides complexed with cationic liposomes were ineffective. Neuroprotection by p53 antisense oligonucleotides in anionic liposomes was comparable with that by glutamate receptor antagonists and a chemical inhibitor of p53. Anionic liposomes were also capable of delivering plasmids and inducing transgene expression in neurons. Anionic liposome-mediated internalization of Cy3 labeled oligonucleotides by neurons and several other cell lines demonstrated the universal applicability of this vector. PMID- 11406619 TI - A positive regulatory role for Cbl family proteins in tumor necrosis factor related activation-induced cytokine (trance) and CD40L-mediated Akt activation. AB - Tumor necrosis factor (TNF)-related activation-induced cytokine (TRANCE) is a TNF family member essential for osteoclast differentiation, and it induces the activation and survival of osteoclasts and mature dendritic cells. We recently demonstrated that TRANCE activates Akt via a mechanism involving TRANCE receptor (TRANCE-R)/RANK, TRAF6, and c-Src. Here, we show that TRANCE-R and CD40 recruit TRAF6, Cbl family-scaffolding proteins, and the phospholipid kinase phosphatidylinositol 3-kinase in a ligand-dependent manner. The recruitment of Cbl-b and c-Cbl to TRANCE-R is dependent upon the activity of Src-family kinases. TRANCE and CD40L-mediated Akt activation is defective in Cbl-b -/- dendritic cells, and CD40L-mediated Akt activation is defective in c-Cbl -/- B cells. These findings implicate Cbl family proteins as not only negative regulators of signaling but as positive modulators of TNF receptor superfamily signaling as well. PMID- 11406620 TI - A protein with characteristics of factor H is present on rodent platelets and functions as the immune adherence receptor. AB - Complement-coated particles interact with specific immune adherence receptors (IAR). In primates, this function is served by complement receptor 1 (CR1) on erythrocytes. In contrast, rodent platelets bear IAR distinct from CR1, the identity of which was studied here. A 150-kDa C3b-binding protein was isolated from rat platelets, which had immunochemical and biochemical identity to plasma factor H. Immunofluorescence microscopy and flow cytometry demonstrated that factor H was present on the surface of rat and mouse platelets, which could be removed by treatment with neuraminidase. Sheep erythrocytes bearing C3b underwent immune adherence with rat and mouse platelets, which was blocked with anti-factor H F(ab')(2) antibodies, but not with antibodies binding to the complement regulator, Crry, on the platelet surface. By reverse transcription-polymerase chain reaction using rat platelet RNA and primers designed from mouse factor H, a 472-base pair product was generated that was identical in sequence to that produced from rat liver RNA. The translated protein product was 85% similar to mouse liver factor H. The 3'-nucleotide sequence from platelets predicted a soluble factor H protein. By Northern analysis, liver and platelets had identically sized factor H mRNA. Thus, rat and mouse platelets have a membrane protein with characteristics of factor H that is linked via sialic acid residues and functions as the IAR. Whether platelet factor H is acquired by passive adsorption from sera and/or is produced by platelets remains to be determined. PMID- 11406621 TI - Cross-talk between caveolae and glycosylphosphatidylinositol-rich domains. AB - Most mammalian cells have in their plasma membrane at least two types of lipid microdomains, non-invaginated lipid rafts and caveolae. Glycosylphosphatidylinositol (GPI)-anchored proteins constitute a class of proteins that are enriched in rafts but not caveolae at steady state. We have analyzed the effects of abolishing GPI biosynthesis on rafts, caveolae, and cholesterol levels. GPI-deficient cells were obtained by screening for resistance to the pore-forming toxin aerolysin, which uses this class of proteins as receptors. Despite the absence of GPI-anchored proteins, mutant cells still contained lipid rafts, indicating that GPI-anchored proteins are not crucial structural elements of these domains. Interestingly, the caveolae-specific membrane proteins, caveolin-1 and 2, were up-regulated in GPI-deficient cells, in contrast to flotillin-1 and GM1, which were expressed at normal levels. Additionally, the number of surface caveolae was increased. This effect was specific since recovery of GPI biosynthesis by gene recomplementation restored caveolin expression and the number of surface caveolae to wild type levels. The inverse correlation between the expression of GPI-anchored proteins and caveolin 1 was confirmed by the observation that overexpression of caveolin-1 in wild type cells led to a decrease in the expression of GPI-anchored proteins. In cells lacking caveolae, the absence of GPI-anchored proteins caused an increase in cholesterol levels, suggesting a possible role of GPI-anchored proteins in cholesterol homeostasis, which in some cells, such as Chinese hamster ovary cells, can be compensated by caveolin up-regulation. PMID- 11406622 TI - In vivo and in vitro regulation of sterol 27-hydroxylase in the liver during the acute phase response. potential role of hepatocyte nuclear factor-1. AB - The host response to infection is associated with several alterations in lipid metabolism that promote lipoprotein production. These changes can be reproduced by lipopolysaccharide (LPS) administration. LPS stimulates hepatic cholesterol synthesis and suppresses the conversion of cholesterol to bile acids. LPS down regulates hepatic cholesterol 7alpha-hydroxylase, the rate-limiting enzyme in the classic pathway of bile acid synthesis. We now demonstrate that LPS markedly decreases the activity of sterol 27-hydroxylase, the rate-limiting enzyme in the alternate pathway of bile acid synthesis, in the liver of Syrian hamsters. Moreover, LPS progressively decreases hepatic sterol 27-hydroxylase mRNA levels by 75% compared with controls over a 24-h treatment period. LPS also decreases oxysterol 7alpha-hydroxylase mRNA levels in mouse liver. In vitro studies in HepG2 cells demonstrate that tumor necrosis factor and interleukin (IL)-1 decrease sterol 27-hydroxylase mRNA levels by 48 and 80%, respectively, whereas IL-6 has no such effect. The IL-1-induced decrease in sterol 27-hydroxylase mRNA expression occurs early, is sustained for 48 h, and requires very low doses. In vivo IL-1 treatment also lowers hepatic sterol 27-hydroxylase mRNA levels in Syrian hamsters. Studies investigating the molecular mechanisms of LPS-induced decrease in sterol 27-hydroxylase show that LPS markedly decreases mRNA and protein levels of hepatocyte nuclear factor-1 (HNF-1), a transcription factor that regulates sterol 27-hydroxylase, in the liver. Moreover, LPS decreases the binding activity of HNF-1 by 70% in nuclear extracts in hamster liver, suggesting that LPS may down-regulate sterol 27-hydroxylase by decreasing the binding of HNF 1 to its promoter. Coupled with our earlier studies on cholesterol 7alpha hydroxylase, these data indicate that LPS suppresses both the classic and alternate pathways of bile acid synthesis. A decrease in bile acid synthesis in liver would reduce cholesterol catabolism and thereby contribute to the increase in hepatic lipoprotein production that is induced by LPS and cytokines. PMID- 11406623 TI - Nitric oxide activates the beta 2 subunit of soluble guanylyl cyclase in the absence of a second subunit. AB - Previously characterized mammalian soluble guanylyl cyclases form alpha/beta heterodimers that can be activated by the gaseous messenger, nitric oxide, and the novel guanylyl cyclase modulator YC-1. Four mammalian subunits have been cloned named alpha(1), beta(1), alpha(2), and beta(2). The alpha(1)/beta(1) and alpha(2)/beta(1) heterodimeric enzyme isoforms have been rigorously characterized. The role of the beta(2) subunit has remained elusive. Here we isolate a novel variant of this subunit and show that the beta(2) subunit does not need to form heterodimers for catalytic activity because enzyme activity can be measured when it is expressed alone in Sf9 cells. In analogy to the beta(3) subunit recently isolated from the insect Manduca sexta, activity was dependent on the presence of 4 mm free Mn(2+). The EC(50) values for the NO-donor diethylamine/NO were shifted to the left by 1 order of magnitude as compared with the alpha(1)/beta(1) heterodimeric form. In the presence of the detergent Tween, NO sensitivity of beta(2) was abolished, but the enzyme could be activated by protoporphyrin IX, indicating removal of a prosthetic heme group and exchange for the heme precursor. We suggest that the beta(2) subunit is the first mammalian NO sensitive guanylyl cyclase lacking a heterodimeric structure. PMID- 11406624 TI - Sequence analysis of heparan sulfate epitopes with graded affinities for fibroblast growth factors 1 and 2. AB - Proteins that belong to the fibroblast growth factor (FGF) family regulate proliferation, migration, and differentiation of many cell types. Several FGFs, including the prototype factors FGF-1 and FGF-2, depend on interactions with heparan sulfate (HS) proteoglycans for activity. We have assessed tissue-derived HS fragments for binding to FGF-1 and FGF-2 to identify the authentic saccharide motifs required for interactions. Sequence information on a range of N-sulfated HS octasaccharides spanning from low to high affinity for FGF-1 was obtained. All octasaccharides with high affinity for FGF-1 (> or =0.5 m NaCl required for elution) contained an internal IdoUA(2-OSO(3))-GlcNSO(3)(6-OSO(3))-IdoUA(2 OSO(3))-trisaccharide motif. Octasaccharides with a higher overall degree of sulfation but lacking the specific trisaccharide motif showed lower affinity for FGF-1. FGF-2 was shown to bind to a mono-O-sulfated HS 6-mer carrying a single internal IdoUA(2-OSO(3))-unit. However, a di-O-sulfated -IdoUA(2-OSO(3)) GlcNSO(3)-IdoUA(2-OSO(3))-trisaccharide sequence within a HS 8-mer gave stronger binding. These findings show that not only the number but also the positions of individual sulfate groups determine affinity of HS for FGFs. Our findings support the notion that FGF-dependent processes can be modulated in vivo by regulated expression of distinct HS sequences. PMID- 11406625 TI - Saposin D solubilizes anionic phospholipid-containing membranes. AB - Saposin (Sap) D is a late endosomal/lysosomal small protein, generated together with three other similar proteins, Sap A, B, and C, from the common precursor, prosaposin. Although the functions of saposins such as Sap B and C are well known (Sap B promotes the hydrolysis of sulfatides and Sap C that of glucosylceramide), neither the physiological function nor the mechanism of action of Sap D are yet fully understood. We previously found that a dramatic increase of Sap D superficial hydrophobicity, occurring at the low pH values characteristic of the late endosomal/lysosomal environment, triggers the interaction of the saposin with anionic phospholipid-containing vesicles. We have presently found that, upon lipid binding, Sap D solubilizes the membranes, as shown by the clearance of the vesicles turbidity. The results of gel filtration, density gradient centrifugation, and negative staining electron microscopy demonstrate that this effect is due to the transformation of large vesicles to smaller particles. The solubilizing effect of Sap D is highly dependent on pH, the lipid/saposin ratio, and the presence of anionic phospholipids; small variations in each of these conditions markedly influences the activity of Sap D. The present study documents the interaction of Sap D with membranes as a complex process. Anionic phospholipids attract Sap D from the medium; when the concentration of the saposin on the lipid surface reaches a critical value, the membrane breaks down into recombinant small particles enriched in anionic phospholipids. Our results suggest that the role played by Sap D is more general than promoting sphingolipid degradation, e.g. the saposin might also be a key mediator of the solubilization of intralysosomal/late endosomal anionic phospholipid-containing membranes. PMID- 11406626 TI - Identification and characterization of the tRNA:Psi 31-synthase (Pus6p) of Saccharomyces cerevisiae. AB - To characterize the substrate specificity of the putative RNA:pseudouridine (Psi) synthase encoded by the Saccharomyces cerevisiae open reading frame (ORF) YGR169c, the corresponding gene was deleted in yeast, and the consequences of the deletion on tRNA and small nuclear RNA modification were tested. The resulting DeltaYGR169c strain showed no detectable growth phenotype, and the only difference in Psi formation in stable cellular RNAs was the absence of Psi at position 31 in cytoplasmic and mitochondrial tRNAs. Complementation of the DeltaYGR169c strain by a plasmid bearing the wild-type YGR169c ORF restored Psi(31) formation in tRNA, whereas a point mutation of the enzyme active site (Asp(168)-->Ala) abolished tRNA:Psi(31)-synthase activity. Moreover, recombinant His(6)-tagged Ygr169 protein produced in Escherichia coli was capable of forming Psi(31) in vitro using tRNAs extracted from the DeltaYGR169c yeast cells as substrates. These results demonstrate that the protein encoded by the S. cerevisiae ORF YGR169c is the Psi-synthase responsible for modification of cytoplasmic and mitochondrial tRNAs at position 31. Because this is the sixth RNA:Psi-synthase characterized thus far in yeast, we propose to rename the corresponding gene PUS6 and the expressed protein Pus6p. Finally, the cellular localization of the green fluorescent protein-tagged Pus6p was studied by functional tests and direct fluorescence microscopy. PMID- 11406627 TI - Directional resolution of synthetic holliday structures by the Cre recombinase. AB - The Cre recombinase of bacteriophage P1 cleaves its target site, loxP, in a defined order. Recombination is initiated on one pair of strands to form a Holliday intermediate, which is then resolved by cleavage and exchange of the other pair of strands to yield recombinant products. To investigate the influence of the loxP sequence on the directionality of resolution, we constructed synthetic Holliday (chi) structures containing either wild-type or mutant lox sites. We found that Cre preferentially resolved the synthetic wild-type chi structures on a particular pair of strands. The bias in the direction of resolution was dictated by the asymmetric loxP sequence since the resolution bias was abolished with symmetric lox sites. Systematic substitutions of the loxP site revealed that the bases immediately 5' to the scissile phosphodiester bonds were primarily responsible for the directionality of resolution. Interchanging these base pairs was sufficient to reverse the resolution bias. The Cre-lox co-crystal structures show that Lys(86) makes a base-specific contact with guanine immediately 5' to one of the scissile phosphates. Substituting Lys(86) with alanine resulted in a reduction of the resolution bias, indicating that this amino acid is important for establishing the bias in resolution. PMID- 11406628 TI - Nucleolytic cleavage of the mixed lineage leukemia breakpoint cluster region during apoptosis. AB - VP-16 (etoposide) has recently been shown to induce topoisomerase II (TOP2) mediated DNA cleavage within the mixed lineage leukemia (MLL) breakpoint cluster region (bcr), suggesting a role of TOP2 in MLL gene rearrangement. In our current studies, we have compared the induction of DNA cleavage within the MLL bcr in different cell lines after treatment with various anticancer drugs. All anticancer drugs tested including VP-16 (a TOP2-directed drug), camptothecin (a topoisomerase I-directed drug), 5-fluorouracil and methotrexate (antimetabolites), and vinblastine (a microtubule inhibitor) induced the same site-specific cleavage within the MLL bcr. This cleavage was shown to be nuclease mediated but not TOP2-mediated by the following observations: 1) drug-induced cleavage within the MLL bcr was not protein-linked; 2) unlike TOP2-mediated cleavage, drug-induced DNA cleavage within the MLL bcr was kinetically slow and coincided with the formation of the apoptotic nucleosomal DNA ladder; 3) drug induced cleavage within the MLL bcr was unaffected in cells with reduced nuclear TOP2; and 4) drug-induced cleavage within the MLL bcr was abolished by the caspase inhibitor, Z-Asp(OCH(3))-Glu(OCH(3))-Val-Asp(OCH(3))-FMK. The possibility that an apoptotic nuclease may be involved in cleavage of the MLL bcr and MLL gene translocation is discussed. PMID- 11406629 TI - Heterogeneous RNA-binding protein M4 is a receptor for carcinoembryonic antigen in Kupffer cells. AB - Here we report the isolation of the recombinant cDNA clone from rat macrophages, Kupffer cells (KC) that encodes a protein interacting with carcinoembryonic antigen (CEA). To isolate and identify the CEA receptor gene we used two approaches: screening of a KC cDNA library with a specific antibody and the yeast two-hybrid system for protein interaction using as a bait the N-terminal part of the CEA encoding the binding site. Both techniques resulted in the identification of the rat heterogeneous RNA-binding protein (hnRNP) M4 gene. The rat ortholog cDNA sequence has not been previously described. The open reading frame for this gene contains a 2351-base pair sequence with the polyadenylation signal AATAAA and a termination poly(A) tail. The mRNA shows ubiquitous tissue expression as a 2.4-kilobase transcript. The deduced amino acid sequence comprised a 78-kDa membrane protein with 3 putative RNA-binding domains, arginine/methionine/glutamine-rich C terminus and 3 potential membrane spanning regions. When hnRNP M4 protein is expressed in pGEX4T-3 vector system in Escherichia coli it binds (125)I-labeled CEA in a Ca(2+)-dependent fashion. Transfection of rat hnRNP M4 cDNA into a non-CEA binding mouse macrophage cell line p388D1 resulted in CEA binding. These data provide evidence for a new function of hnRNP M4 protein as a CEA-binding protein in Kupffer cells. PMID- 11406630 TI - A pair of fluorescent resonance energy transfer-based probes for tyrosine phosphorylation of the CrkII adaptor protein in vivo. AB - An adaptor protein, CrkII, which is involved in a variety of signaling cascades such as cell growth, migration, and apoptosis, becomes phosphorylated on Tyr(221) upon stimulation. Here, we report on a fluorescent resonance energy transfer based sensor, which consists of CrkII sandwiched with cyan- and yellow-emitting variants of green fluorescent protein. This protein enabled us to monitor rapid and transient phosphorylation of CrkII upon epidermal growth factor stimulation in a living cell. However, rapid diffusion of the probes prevented us from specifying where the phosphorylation started within the cell. To overcome this problem, we fused the CAAX box of Ki-Ras to the carboxyl terminus of this probe and restricted its localization mostly to the plasma membrane. With this modified probe, we found that epidermal growth factor-induced phosphorylation of CrkII was initiated at the peripheral plasma membrane, moving toward the center of the cell. Moreover, this CAAX box-fused probe showed improvement in sensitivity and time resolution of the monitoring of CrkII phosphorylation. Thus, this pair of CrkII probes visualizes dynamic changes in the total and local levels of the tyrosine phosphorylation of CrkII in a living cell. PMID- 11406631 TI - Impaired hearing in mice lacking aquaporin-4 water channels. AB - A role for aquaporins (AQPs) in hearing has been suggested from the specific expression of aquaporins in inner ear and the need for precise volume regulation in epithelial cells involved in acoustic signal transduction. Using mice deficient in selected aquaporins as controls, we localized AQP1 in fibrocytes in the spiral ligament and AQP4 in supporting epithelial cells (Hensen's, Claudius, and inner sulcus cells) in the organ of Corti. To determine whether aquaporins play a role in hearing, auditory brain stem response (ABR) thresholds were compared in wild-type mice and transgenic null mice lacking (individually) AQP1, AQP3, AQP4, and AQP5. In 4-5-week-old mice in a CD1 genetic background, ABR thresholds in response to a click stimulus were remarkably increased by >12 db in AQP4 null mice compared with wild-type mice (p < 0.001), whereas ABR thresholds were not affected by AQP1, AQP3, or AQP5 deletion. In a C57/bl6 background, nearly all AQP4 null mice were deaf, whereas ABRs could be elicited in wild-type controls. ABRs in AQP4 null CD1 mice measured in response to tone bursts (4-20 kHz) indicated a frequency-independent hearing deficit. Light microscopy showed no differences in cochlear morphology of wild-type versus AQP4 null mice. These results provide the first direct evidence that an aquaporin water channel plays a role in hearing. AQP4 may facilitate rapid osmotic equilibration in epithelial cells in the organ of Corti, which are subject to large K(+) fluxes during mechano-electric signal transduction. PMID- 11406632 TI - Reengineering granulocyte colony-stimulating factor for enhanced stability. AB - Granulocyte colony-stimulating factor is a long-chain cytokine that has both biological and therapeutic applications. It is involved in the production and maturation of neutrophilic progenitor cells and neutrophils and is administered to stimulate the production of white blood cells to reduce the risk of serious infection in immunocompromised patients. We have reengineered granulocyte colony stimulating factor to improve the thermodynamic stability of the protein, focusing on enhancing the alpha-helical propensity of residues in the antiparallel 4-helix bundle of the protein. These redesigns resulted in proteins with substantially enhanced stability while retaining wild-type levels of biological activity, measured as the ability of the reengineered proteins to stimulate the proliferation of murine myeloid cells transfected with the granulocyte colony-stimulating factor receptor. PMID- 11406633 TI - Amelogenin-deficient mice display an amelogenesis imperfecta phenotype. AB - Dental enamel is the hardest tissue in the body and cannot be replaced or repaired, because the enamel secreting cells are lost at tooth eruption. X-linked amelogenesis imperfecta (MIM 301200), a phenotypically diverse hereditary disorder affecting enamel development, is caused by deletions or point mutations in the human X-chromosomal amelogenin gene. Although the precise functions of the amelogenin proteins in enamel formation are not well defined, these proteins constitute 90% of the enamel organic matrix. We have disrupted the amelogenin locus to generate amelogenin null mice, which display distinctly abnormal teeth as early as 2 weeks of age with chalky-white discoloration. Microradiography revealed broken tips of incisors and molars and scanning electron microscopy analysis indicated disorganized hypoplastic enamel. The amelogenin null phenotype reveals that the amelogenins are apparently not required for initiation of mineral crystal formation but rather for the organization of crystal pattern and regulation of enamel thickness. These null mice will be useful for understanding the functions of amelogenin proteins during enamel formation and for developing therapeutic approaches for treating this developmental defect that affects the enamel. PMID- 11406634 TI - Human transforming growth factor-beta activates a receptor serine/threonine kinase from the intravascular parasite Schistosoma mansoni. AB - The biology of the helminth parasite Schistosoma mansoni is closely integrated with that of its mammalian host. SmRK1, a divergent type I transforming growth factor-beta (TGF-beta) receptor of unknown ligand specificity, was previously identified as a candidate for a receptor that allows schistosomes to respond to host-derived growth factors. The TGF-beta family includes activin, bone morphogenetic proteins (BMPs), and TGF-beta, all of which can play crucial roles in metazoan development. The downstream signaling protein of receptors that respond to TGF-beta and activin is Smad2, whereas the receptors that respond to BMPs signal via Smad1. When a constitutively active mutant of SmRK1 was overexpressed with either schistosome Smad1 (SmSmad1) or SmSmad2, a receptor dependent modulation of SmSmad phosphorylation and luciferase reporter activity occurred only with SmSmad2. To evaluate potential ligand activators of SmRK1, a chimeric receptor containing the extracellular domain of SmRK1 joined to the intracellular domain of the human type I TGF-beta receptor was used. The chimeric receptor bound radiolabeled TGF-beta and could activate a luciferase reporter gene in response to both TGF-beta 1 and TGF-beta 3 but not BMP7. Confirmatory results were obtained using full-length SmRK1. These experiments implicate TGF beta as a ligand for SmRK1 and as a potential host-derived regulator of parasite growth and development. PMID- 11406635 TI - De novo-designed peptide transforms Golgi-specific lipids into Golgi-like nanotubules. AB - Cellular organelles, such as the Golgi apparatus and the endoplasmic reticulum, adopt characteristic structures depending on their function. While the tubular shapes of these structures result from complex protein-lipid interactions that are not fully understood, some fundamental machinery must be required. We show here that a de novo-designed 18-mer amphipathic alpha-helical peptide, Hel 13-5, transforms spherical liposomes made from a Golgi-specific phospholipid mixture into nanotubules on the scale of and resembling the shape of the nanotubules that form the Golgi apparatus. Furthermore, we show that that the size and the shape of such nanotubules depend on lipid composition and peptide properties such as length and the ratio of hydrophobic to hydrophilic amino acids. Although the question of precisely how nature engineers organellar membranes remains unknown, our simple novel system provides a basic set of tools to begin addressing this question. PMID- 11406636 TI - Terminal deoxynucleotidyl transferase indiscriminately incorporates ribonucleotides and deoxyribonucleotides. AB - Terminal deoxynucleotidyl transferase (TdT) catalyzes the condensation of deoxyribonucleotides on 3'-hydroxyl ends of DNA strands in a template-independent manner and adds N-regions to gene segment junctions during V(D)J recombination. Although TdT is able to incorporate a few ribonucleotides in vitro, TdT discrimination between ribo- and deoxyribonucleotides has never been studied. We found that TdT shows only a minor preference for incorporation of deoxyribonucleotides over ribonucleotides on DNA strands. However, incorporation of ribonucleotides alone or in the presence of deoxyribonucleotides generally leads to premature chain termination, reflecting an impeded accommodation of ribo or mixed ribo/deoxyribonucleic acid substrates by TdT. An essential catalytic aspartate in TdT was identified, which is a first step toward understanding the apparent lack of sugar discrimination by TdT. PMID- 11406637 TI - Evidence for functional release of endogenous opioids in the locus ceruleus during stress termination. AB - Endogenous opioids target noradrenergic locus ceruleus (LC) neurons and potently inhibit LC activity. Nonetheless, it has been difficult to demonstrate functional regulation of the LC-noradrenergic system by endogenous opioids because of the lack of effect of opiate antagonists. The present findings provide evidence that endogenous opioids regulate LC neuronal activity during the termination of a stressor. LC neuronal discharge was recorded from halothane-anesthetized rats before, during, and after hypotensive stress elicited by intravenous nitroprusside infusion. In naive rats, mean arterial blood pressure was temporally correlated with LC activity such that hypotension was associated with increased LC discharge and a return to the normotensive state was associated with a decrease in LC discharge below pre-stress values. After microinfusion of an antagonist of the stress neuropeptide corticotropin-releasing factor (CRF) into the LC, the increase in LC discharge associated with hypotension was prevented, whereas LC inhibition associated with termination of the challenge occurred at an earlier time and was of a greater magnitude. In contrast, microinfusion of naloxone into the LC completely abolished LC inhibition associated with termination of the stressor. Naloxone microinfusion did not prevent LC inhibition associated with hypertension produced by intravenous vasopressin administration, suggesting that endogenous opioids may be selectively engaged during the termination of hypotensive stress. These results provide evidence for a functional release of endogenous opioids within the LC. This action of endogenous opioids may serve to counterbalance excitatory effects of CRF on the LC norepinephrine system, thereby limiting its activation by stress. PMID- 11406638 TI - Spontaneous remission of paroxysmal dystonia coincides with normalization of entopeduncular activity in dt(SZ) mutants. AB - Recent studies have shown a dramatically decreased spontaneous discharge rate of entopeduncular neurons in a unique animal model of idiopathic paroxysmal dystonia, the dt(sz) mutant hamster. These changes were found in animals at the age at which the most marked expression of dystonia is usually observed. In this rodent model, the age-dependent disappearance of stress-inducible dystonic attacks at an age of approximately 10 weeks allows investigations of the relevance of pathophysiological changes for the occurrence of dystonia by ontogenetic studies. Therefore, we examined the entopeduncular activity by extracellular single unit recordings in groups of dt(sz) mutants and nondystonic control hamsters at 17-22 weeks of age. In contrast to recent findings, after the complete remission of dystonia, the mean discharge rate of entopeduncular neurons in dt(sz) mutants (28.1 +/- 1.2 spikes/sec) was similar to that of age-matched nondystonic control hamsters (30.8 +/- 0.9 spikes/sec). Thus, the disappearance of paroxysmal dystonia is accompanied by a normalization of the entopeduncular activity in dt(sz) mutants. The present data clearly demonstrate the fundamental importance of a decreased basal ganglia output for the expression of paroxysmal dystonia. PMID- 11406640 TI - Mitochondrial dna depletion, oxidative stress, and mutation: mechanisms of dysfunction from nucleoside reverse transcriptase inhibitors. PMID- 11406641 TI - Her-2/neu gene amplification, elevated mRNA expression, and protein overexpression in the metaplasia-dysplasia-adenocarcinoma sequence of Barrett's esophagus. AB - SUMMARY: The importance of alterations of the Her-2/neu oncogene in the tumorigenesis of Barrett's adenocarcinoma (BCA) is discussed controversially. In the present study, we evaluated for the first time the Her-2/neu status in the metaplasia-dysplasia-adenocarcinoma sequence of BCA simultaneously at the DNA, mRNA, and protein level using resection specimens of 25 patients. The locus specific Her-2/neu gene status was quantified by performing fluorescence in situ hybridization, and information about the ploidy status of chromosome 17 was obtained. Tissue sections from the same areas were used for quantitative RT-PCR (TaqMan RT-PCR) of laser-microdissected tumor cells and for immunohistochemistry to quantify Her-2/neu mRNA and oncoprotein expression. Her-2/neu gene amplification was observed in 35% of BCA, and all of these samples showed strong overexpression of both mRNA and oncoprotein. A polysomy 17 without Her-2/neu gene amplification was observed in 52% of BCA, showing a normal or moderately elevated mRNA expression and no or weak immunopositivity. From 13 areas of high-grade dysplasia (HGD) we found four to be amplified for the Her-2/neu locus, whereas five showed a polysomy 17. All four samples of HGD areas with Her-2/neu gene amplification displayed mRNA and strong oncoprotein overexpression; however, lower mRNA levels were seen than in the amplified BCA areas. None of the samples with low-grade dysplasia (LGD) showed a locus-specific Her-2/neu amplification, but polysomy 17 was present in four of eight cases. No changes were detected in BCA-associated intestinal metaplasia and squamous epithelium. In summary, only a locus-specific Her-2/neu gene amplification was associated with strong mRNA overexpression and strong membranous Her-2/neu immunostaining in BCA and HGD. A chromosome 17 polysomy, as found in the majority of BCA, led to no or weak mRNA overexpression and no or weak immunopositivity. In the metaplasia-dysplasia adenocarcinoma sequence, a chromosome 17 polysomy without Her-2/neu gene amplification was already present in LGD. This may be a result of an early polyploidization, preceding the later genetic events, such as Her-2/neu gene amplification in HGD and BCA. PMID- 11406642 TI - Molecular analysis of the 9p21 locus and p53 genes in Ewing family tumors. AB - SUMMARY: The EWS-ETS rearrangements, and their respective fusion gene products, are specifically associated with histopathologically Ewing family tumors (EFT). These translocations are implicated in generating malignant transformation of EFT, but the presence of additional genetic alterations must be considered in the pathogenesis of such tumors. We analyzed 26 samples (biopsies and/or nude mice xenotransplants) collected from 19 patients with an EFT to determine whether molecular and cytogenetic alterations of the G(1)/S checkpoint genes are implicated in the pathogenesis of EFT. We found inactivating p53 mutations in three (16%) cases, which correlated with a loss of p21(WAF1/Cip1) expression and with a monosomy of chromosome 17 in two cases. Homozygous deletion of the p16(INK4A)/p14(ARF) gene was detected in four (21%) cases, three with codeletion of the p15(INK4B) gene and with chromosome 9 abnormalities. In all of these cases, expression of the implicated genes was absent. Hypermethylation of the p16(INK4A) and p15(INK4B) genes was detected in two (10%) and three (16%) cases, respectively, and was correlated with a low level of gene expression. Neither cyclin D1, nor MDM2 and CDK4 amplification was observed. Kaplan-Meier analysis showed that patients with tumors carrying homozygous deletion of the 9p21 locus, or point mutations of the p53 gene, had poorer outcomes than those without these molecular alterations (p = 0.005). In conclusion, 58% (11 of 19) of the analyzed patients showed genetic or epigenetic alterations in either the 9p21 locus or p53 tumor suppressor genes, defining a subgroup of patients with poor clinical outcome. This fact points to an important role of the G(1)/S cell cycle checkpoint dysregulation in the pathogenesis of EFT. PMID- 11406643 TI - Increased expression of FGF-8 isoforms and FGF receptors in human premalignant prostatic intraepithelial neoplasia lesions and prostate cancer. AB - SUMMARY: Fibroblast growth factor 8 (FGF-8) is implicated in growth of prostate cancer. Alternative splicing of the human FGF-8 gene potentially allows coding for four protein isoforms (a, b, e, and f). These isoforms differ in their binding to FGF receptors (FGFR) and in their mitogenic and transforming capacity in transfection assays. Here, we used RT-PCR and immunohistochemistry to study the expression of FGF-8 and FGFR isoforms in human prostate cancer (n = 31). Nonmalignant prostate specimens from cystoprostatectomies (n = 24) were examined as controls. Most prostate cancer samples and some control prostates also contained prostatic intraepithelial neoplasia (PIN) lesions. FGF-8a and e were expressed at significantly higher frequencies in prostate cancer (FGF-8a, 55%; FGF-8e, 45%) than in control samples (FGF-8a, 17%, p = 0.0052; FGF-8e, 8%, p = 0.0031). On the contrary, FGF-8b was found at an equal frequency in prostate cancer (55%) and in control prostates (50%). Furthermore, a combination of two or three FGF-8 isoforms (a, b, and/or e) was also expressed at a higher frequency in prostate cancer than in control samples (45% and 8%, respectively, p = 0.0031). Immunohistochemistry with an antibody recognizing all FGF-8 isoforms was more strongly immunoreactive in prostate cancer cells and PIN lesions than in normal type epithelium. The receptor splicing variants FGFR1IIIc and FGFR2IIIc, which are activated by FGF-8, were found both in prostate cancer and control samples. Interestingly, immunoreactivity for FGFR1 and FGFR2 was much stronger in prostate cancer cells and PIN than in normal epithelium. These results demonstrate, for the first time, that FGF-8 isoforms and their receptors FGFR1IIIc and FGFR2IIIc are expressed at an increased level not only in prostate cancer but also in premalignant PIN lesions. These data suggest that FGF-8 may have an important autocrine role in the development of human prostate cancer. In addition to FGF 8b, the FGF-8 isoforms a and e may be involved in this process. PMID- 11406644 TI - Enrichment of mutant KRAS alleles in pancreatic juice by subtractive iterative polymerase chain reaction. AB - SUMMARY: The detection of mutant tumor genes holds great promise for an early diagnosis of primary tumors and residual malignant disease. When few tumor cells are present with an excess of nonmalignant cells of the same lineage, the excess of wild-type alleles over mutant tumor alleles presents an analytical problem. The subtractive iterative PCR (siPCR) assay presents a new approach to solving this problem. To achieve an enrichment of mutant alleles, wild-type alleles are removed by differential hybridization to complementary oligonucleotides spanning the region of the gene in which point mutations are expected. The nonbound fraction is reamplified by PCR. By iterating this process, mutant alleles can be detected in the presence of an excess of wild-type alleles with high sensitivity. To prove the feasibility of siPCR, pancreatic juice samples were analyzed for KRAS mutations. Pancreatic juice obtained from patients with pancreatic carcinoma or chronic pancreatitis during endoscopic retrograde cholangiopancreatography was analyzed for point mutations in codons 12 and 13 of the KRAS gene. In each of six samples from tumor patients, mutations in codon 12 were detected. One of nine samples from patients with chronic pancreatitis scored positive. PMID- 11406645 TI - Rb and TP53 pathway alterations in sporadic and NF1-related malignant peripheral nerve sheath tumors. AB - SUMMARY: Karyotypic complexities associated with frequent loss or rearrangement of a number of chromosome arms, deletions, and mutations affecting the TP53 region, and molecular alterations of the INK4A gene have been reported in sporadic and/or neurofibromatosis type I (NF1)-related malignant peripheral nerve sheath tumors (MPNSTs). However, no investigations addressing possible different pathogenetic pathways in sporadic and NF1-associated MPNSTs have been reported. This lack is unexpected because, despite similar morphologic and immunophenotypic features, NF1-related cases are, by definition, associated with NF1 gene defects. Thus, we investigated the occurrence of TP53 and p16(INK4A) gene deregulation and the presence of microsatellite alterations at markers located at 17p, 17q, 9p21, 22q, 11q, 1p, or 2q loci in MPNSTs and neurofibromas either related (14 cases) or unrelated (14 cases) to NF1. Our results indicate that, in MPNSTs, p16(INK4A) inactivation almost equally affects both groups. However, TP53 mutations and loss of heterozygosity involving the TP53 locus (43% versus 9%), and p53 wild type overexpression, related or not to mdm2 overexpression (71% versus 25%), seem to mainly be restricted to sporadic MPNSTs. In NF1-associated MPNSTs, our microsatellite results are consistent with the occurrence of somatic inactivation by loss of heterozygosity of the second NF1 allele. PMID- 11406646 TI - Immunohistochemical distribution and quantitative biochemical detection of advanced glycation end products in fetal to adult rats and in rats with streptozotocin-induced diabetes. AB - SUMMARY: We used immunohistochemical methods and four monoclonal antibodies for specific molecular structures of advanced glycation end products (AGE)-6D12, KNH 30, 1F6, and 2A2-to examine localization of AGE in fetal, young, and adult rats, and rats with streptozotocin-induced diabetes. 6D12 recognized N(epsilon) (carboxymethyl)lysine (CML); KNH-30, N(epsilon)-(carboxyethyl)lysine (CEL); and 1F6, fluorolink. The epitope of 2A2 is as yet unknown. Immunoreactivities for these monoclonal antibodies were found in various organs and tissues in postnatal and adult rats, and accumulation increased with aging. In the fetuses, AGE structures were detected at 10 fetal days, and their accumulation increased during ontogeny. Reversed-phase high-performance liquid chromatography revealed CML in fetuses at 13 fetal days and in lungs of 28-week-old rats. In various organs and tissues of fetal, young, and adult rats, CML, CEL, 2A2-positive AGE, and fluorolink accumulated, in that order, which suggests that the accumulation of CML, a nonfluorescent/noncross-linked AGE, occurs earlier than accumulation of fluorolink, a fluorescent/cross-linked AGE. In diabetic rats, hepatocytes, splenic macrophages, renal glomerular endothelial and mesangial cells, testicular Leydig cells, and erythrocytes showed excessive accumulation of AGE, leading to the pathologic changes characteristic of diabetes mellitus. For the induction of these changes, persistent hyperglycemia and hyperketonemia might be important for acceleration of intracellular AGE accumulation in diabetic rats. Thus, AGE accumulation in tissues and cells occurs not only during aging and in diabetes mellitus but also from an early stage of ontogeny. PMID- 11406647 TI - Doc-2/hDab2 expression is up-regulated in primary pancreatic cancer but reduced in metastasis. AB - SUMMARY: DOC-2/hDab2 (DOC-2) has tumor suppressive functions in ovarian cancer and choriocarcinoma. In these tumors, it negatively influences mitogenic signal transduction of growth factors and blocks ras activity. Pancreatic cancer exhibits a high frequency of K-ras gene mutations; however, it is not known whether DOC-2 expression is altered in these tumors. Therefore, we investigated DOC-2 expression in 22 pancreatic adenocarcinomas and in 6 pancreatic cancer cell lines. Findings in human tumors were compared with normal controls and correlated with clinicopathological data. Additionally, the influence of K-ras on DOC-2 transcription was investigated. Northern blot and Western blot analyses both demonstrated an increase of DOC-2 mRNA and protein levels in primary pancreatic cancers in comparison with normal controls. In situ hybridization showed DOC-2 mRNA expression in the majority of cancer cells of primary tumors, as well as in chronic pancreatitis-like lesions surrounding the cancer mass. Immunohistochemistry mirrored the in situ hybridization findings. In contrast, levels of expression of DOC-2 in lymph node metastases were markedly decreased in comparison with levels in primary tumors. In addition, in 5 metastatic pancreatic cancer cell lines, DOC-2 mRNA and protein levels were low, whereas quantitative RT-PCR demonstrated relatively higher levels in a nonmetastatic pancreatic cancer cell line. In conclusion, DOC-2 is overexpressed in primary pancreatic adenocarcinoma but down-regulated in metastatic disease, suggesting a tumor suppressor function of DOC-2 in the late steps of pancreatic carcinogenesis. PMID- 11406648 TI - Human vasculogenesis ex vivo: embryonal aorta as a tool for isolation of endothelial cell progenitors. AB - SUMMARY: Vasculogenesis, the de novo formation of new blood vessels from undifferentiated precursor cells or angioblasts, has been studied with experimental in vivo and ex vivo animal models, but its mechanism is poorly understood, particularly in humans. We used the aortic ring assay to investigate the angioforming capacity of aortic explants from 11- to 12-week-old human embryos. After being embedded in collagen gels, the aorta rings produced branching capillary-like structures formed by mesenchymal spindle cells that lined a capillary-like lumen and expressed markers of endothelial differentiation (CD31, CD34, von Willebrand factor [vWF], and fms-like tyrosine kinase-1 [Flk 1]/vascular endothelial growth factor receptor 2 [VEGFR2]). The cell linings of these structures showed ultrastructural evidence of endothelial differentiation. The neovascular proliferation occurred primarily in the outer aspects of aortic rings, thus suggesting that the new vessels mainly arose from immature endothelial precursor cells localized in the outer layer of the aortic stroma, ie, a process of vasculogenesis rather than angiogenesis. The undifferentiated mesenchymal cells (CD34+/CD31-), isolated and cultured on collagen-fibronectin, differentiated into endothelial cells expressing CD31 and vWF. Furthermore, the CD34+/CD31+ cells were capable of forming a network of capillary-like structures when cultured on Matrigel. This is the first reported study showing the ex vivo formation of human microvessels by vasculogenesis. Our findings indicate that the human embryonic aorta is a rich source of CD34+/CD31- endothelial progenitor cells (angioblasts), and this information may prove valuable in studies of vascular regeneration and tissue bioengineering. PMID- 11406649 TI - Truncated typeXVII collagen expression in a patient with non-herlitz junctional epidermolysis bullosa caused by a homozygous splice-site mutation. AB - SUMMARY: Type XVII collagen (180-kDa bullous pemphigoid antigen) is a structural component of hemidesmosomes. Mutations in the type XVII collagen gene (COL17A1) have been established to be the molecular basis of non-Herlitz junctional epidermolysis bullosa (JEB-nH), an inherited skin blistering disorder. Here we report for the first time truncated type XVII collagen expression, caused by homozygosity for a COL17A1 donor splice-site mutation (4261+1 g --> c), which was identified by PCR amplification on genomic DNA. By RT-PCR and sequencing of cDNA derived from mRNA from the patient's cultured keratinocytes, we provide evidence of cryptic splicing and exon skipping, most abundantly of exon 52. JEB-nH patients with COL17A1 splice-site mutations resulting in an exon skip often have no immunologically detectable type XVII collagen. However, in our patient with the generalized atrophic benign epidermolysis bullosa subtype, a small amount of type XVII collagen was detectable in the skin, and immunoblotting of cultured keratinocytes revealed that the 180-kDa protein was 10 kDa shorter. We hypothesize that the function of this truncated type XVII collagen polypeptide, which is expressed at low levels, is impaired, explaining the JEB-nH phenotype. PMID- 11406650 TI - Effect of p27 deficiency and rapamycin on intimal hyperplasia: in vivo and in vitro studies using a p27 knockout mouse model. AB - SUMMARY: Rapamycin, an immunosuppressant and antiproliferative agent, reduces intimal hyperplasia after arterial injury in animal models and in a preliminary study in humans. Rapamycin treatment reportedly increases expression of p27, a cyclin-dependent kinase inhibitor. This mechanism was tested using a p27 deficient (p27 -/-) murine model. Aortic smooth muscle cells from wild-type (WT) and p27 -/- mice were isolated and cultured. Cell proliferation, assessed by cell count and (3)H-thymidine incorporation, was inhibited significantly by rapamycin in WT and p27 -/- cells at concentrations of 1 ng/ml, 10 ng/ml, and 100 ng/ml (p < 0.05, versus control). The in vivo effect on intimal hyperplasia was studied in p27 -/- and WT mice after femoral artery transluminal injury. Rapamycin treatment was started 2 days before injury and maintained for 2 weeks (1 mg/kg per 48 hours, ip). No significant differences in intima-to-media ratio were found between WT (1.1 +/- 0.1) and p27 -/- mice (1.0 +/- 0.1) 4 weeks after injury. Rapamycin significantly (p < 0.05) reduced intima-to-media ratios in both WT (0.7 +/- 0.1) and p27 -/- mice (0.5 +/- 0.1), compared with untreated mice. p27 deficiency did not alter the arterial wall proliferative response to injury. The inhibitory effect of rapamycin on intimal hyperplasia occurred via a p27 independent mechanism. The in vitro data showed that this effect was mediated through decreased proliferation and enhanced apoptosis. PMID- 11406651 TI - A multiplex real-time pcr assay for the detection of gene fusions observed in solid tumors. AB - SUMMARY: Specific gene fusions observed in solid tumors are extremely useful diagnostic markers. We report the development of a method based on real-time PCR which enables the detection upon identical PCR conditions of the different fusions specifically observed in Ewing tumors (ET), alveolar rhabdomyosarcoma (ARMS), synovial sarcoma (SS), small round cell desmoplastic tumors (SRCDT), extraskeletal myxoid chondrosarcoma, malignant melanoma of soft parts, congenital fibrosarcoma, and anaplastic large cell lymphoma. A simple assay, based on multiplexing of primers and probes, is described for the routine genetic diagnosis of small round cell tumors of children. It enables the detection of the five EWS-ETS, the two PAX-FKHR, the three SYT-SSX, and the EWS-WT1 fusions of ET, ARMS, SS, and SRCDT, respectively. The sensitivity of this test is high enough to detect all fusions, including the large EWS-FLI-1 transcripts, with the equivalent of 100 tumor cells as a starting material. This multiplex fluorescent analysis of chromosome translocations (MFACT) was validated in comparison with conventional RT-PCR on a series of 79 tumors. A major advantage of this method is that it completely abolishes the manipulation of PCR-products. It, therefore, considerably lowers the risk of cross-contamination linked to carry-over of RT PCR products. It also constitutes an important step toward the complete automation of the detection of cancer-specific gene fusions. PMID- 11406652 TI - Activity-induced expression of common reference genes in individual cns neurons. PMID- 11406653 TI - The role of androgen receptors in the clinical course of nevus sebaceus of Jadassohn. AB - Nevus sebaceus of Jadassohn (NSJ) is a benign, congenital hamartoma that often presents at birth, appears to regress in childhood, and grows during puberty, suggesting possible hormonal control. We studied 18 cases of NSJ from children and adults for immunohistochemical evidence of androgen receptor expression. The lesions were evaluated for location and pattern of immunostaining, and these findings were compared between age groups, sexes, and to androgen receptor expression in normal skin. Androgen receptor positivity was seen in the sebaceous glands, in eccrine glands with and without apocrine change, and rarely in keratinocytes in the sebaceous nevi. There were no significant differences in staining location or pattern between the age groups or sexes. Normal skin showed similar staining in the sebaceous glands but did not show staining of the eccrine glands or keratinocytes. Androgen receptors are present in all epithelial components of NSJ, but there is no change in androgen receptor expression during puberty. PMID- 11406654 TI - Microsatellite instability in double primary cancers of the colorectum and stomach. AB - Little is known about genetic alterations of patients who present multiple primary cancers. We hypothesized that microsatellite instability (MSI) is one of the underlying genetic factors in the development of double primary cancers in colorectal cancer patients. We examined for MSI in 41 colorectal cancer patients who presented with extra-colonic primary cancers consisted of 17 gastric and 24 non-gastric cancers. Coincident MSI+ in tumors of two organs were observed in 3 (17.7%) of 17 patients with colon and stomach cancers and 0 of 24 patients with colon and non-gastric cancers (P =.03). In 17 patients with colon and stomach cancers, 6 (31.6%) of 19 colon cancers and 3 (17.7%) of 17 gastric cancers exhibited MSI+. Among four patients with metachronous colon cancers who were identified within the 41 double primary cancer patients, two patients were associated with the MSI+ phenotype. In summary, the prevalent coincidence of MSI suggests that genetic defect of mismatch repair deficiency may be responsible for a small subset of double primary cancers of the colorectum and stomach. PMID- 11406655 TI - Leptin and leptin receptor mRNA are widely expressed in tumors of adipocytic differentiation. AB - Adipose tissue is the principal source of leptin, a cytokine-like peptide with many biologic functions. Leptin binds to the leptin receptor, present in the hypothalamus and in many other tissues, and modulates energy balance and maintenance of body weight. The expression of leptin and leptin receptor in tumors of adipocytic differentiation has not been previously examined. Because normal adipose tissue is the principal source of leptin and expresses leptin receptor, we hypothesized that tumors of adipose tissue differentiation may also express leptin and/or the long functional form of the leptin receptor (OB-Rb). Leptin and OB-Rb were analyzed by immunohistochemistry, in situ hybridization, RT PCR, and western blotting in 21 lipomas, 2 hibernomas, and 16 liposarcomas. Immunostaining and in situ hybridization showed leptin and OB-Rb mRNA expression in all cases of lipomas, hibernomas, and liposarcomas, including dedifferentiated and pleomorphic liposarcomas. RT-PCR analysis showed leptin and OB-Rb mRNA in both lipomas (n = 5) and liposarcomas (n = 5). Western blotting identified the 16 kDa leptin protein in a lipoma and a liposarcoma. No important difference in the expression of leptin and OB-Rb mRNA was found between lipomas and liposarcomas, although the level of leptin protein was higher in a lipoma than a liposarcoma by western blotting. These results show for the first time that leptin and OB-Rb mRNA are expressed in lipomas, hibernomas, and liposarcomas. The presence of leptin and its receptor may provide new insights into the pathobiology of these tumors. PMID- 11406656 TI - Protein overexpression and gene amplification of c-erb B-2 in pulmonary carcinomas: a comparative immunohistochemical and fluorescence in situ hybridization study. AB - Amplification of the c-er bB-2 gene (located on 17q11.2--12) is accompanied by overexpression of its cell surface receptor product, p185(ERBB2). In pulmonary carcinomas, however, there has been disagreement between the reported frequencies of gene amplification and overexpression. To clarify their relationship, the correlation between the cellular expression of p185(ERBB2) and the level of c-erb B-2 gene amplification was studied. A total of 195 pulmonary carcinomas (182 primary and 13 metastatic) were examined immunohistochemically using a polyclonal antibody, which recognizes the internal domain of the human c-erb B-2 protein, and positive tumors were further examined for the gene amplification by dual color fluorescence in situ hybridization using probes for centromere 17 and 17q11.2--12. By immunohistochemistry, distinct membrane staining was found in an adenocarcinoma, a large cell carcinoma and a metastatic carcinoma from the breast, and cytoplasmic and/or faint membranous staining was observed in 23 non small cell lung carcinomas. It was in the two primaries and the metastatic carcinoma that more than 8-fold amplification of c-erb B-2 was found by fluorescence in situ hybridization. Especially, in the two primary carcinomas, tumor cells had amplified genes with the signals forming one or two clusters, indicating that the amplified gene was present in homogeneously staining regions. Among the 23 tumors, three tumors showed low-level amplification (less than 3 fold), which was differentiated from polysomy 17 found in the other two. In the 30 non-small cell lung carcinomas selected at random from 151 with negative immunostaining, there were five trisomy 17, but no tumors with the gene amplification. This suggests that although c-erb B-2 amplification in pulmonary carcinoma is rare, it occurs in the form of a homogeneously staining region and is thought to control the overexpression of the protein in the cell membrane. New adjuvant therapy using a humanized antibody to the oncoprotein may be beneficial to patients with these tumors. PMID- 11406657 TI - Abdominopelvic sarcoma of perivascular epithelioid cells. Report of four cases in young women, one with tuberous sclerosis. AB - The perivascular epithelioid cell has been proposed to be the unifying proliferating cell type in a number of lesions such as angiomyolipoma, lymphangiomyomatosis, clear cell "sugar" tumor and renal capsuloma. With the exception of rare examples of angiomyolipoma, they are non-metastasizing. We report four examples of a new member of this family of perivascular epithelioid cell neoplasms that occur in abdominopelvic location and show metastatic properties. The patients, all women, were aged 19 to 41 years (mean, 32), and presented with a tumor mass involving the serosa of the ileum, uterus or pelvic cavity. Morphologically, the tumors were composed of sheets of large polygonal cells with glycogen-rich clear or eosinophilic cytoplasm and moderately pleomorphic nuclei, traversed by a delicate vasculature, mimicking clear cell carcinoma. There were areas of coagulative necrosis and occasional mitotic figures. Intracytoplasmic brown pigment was present in two cases. Spindly cells, smooth muscle and fat were absent. Lymphovascular invasion was present in all, lymph node metastasis was documented in two and metastasis to the ovary was present in one case. Two patients developed widespread metastatic disease after 10 and 28 months from diagnosis. One patient showed the clinical signs of tuberous sclerosis. In spite of the epithelial-like appearance, the tumor cells were negative for epithelial markers but were strongly positive with the melanogenesis-related marker HMB45. Another melanogenesis marker (MART-1) was positive in two cases. Other markers including S-100 protein, vimentin, muscle specific actin, desmin and chromogranin A were negative. Thus, these tumors are not readily classifiable in the existing schema of known entities, and show overlapping morpho-phenotypic features of clear cell "sugar" tumor of the lung and epithelioid angiomyolipoma. We consider them as sarcomas composed of a pure population of uncommitted perivascular epithelioid cell, that lack modulation toward smooth muscle or adipose cells. PMID- 11406658 TI - ALK1 and p80 expression and chromosomal rearrangements involving 2p23 in inflammatory myofibroblastic tumor. AB - BACKGROUND: Inflammatory myofibroblastic tumor (IMT) is an uncommon tumor of extrapulmonary and pulmonary tissues with an unpredictable clinical course, occasional recurrences, and rare malignant transformation. Clonal abnormalities with rearrangements of chromosome of 2p23 and the ALK gene have been reported in a few cases. The purpose of this study is to investigate whether these are consistent abnormalities among IMTs or represent a distinct subset. DESIGN: Formalin-fixed, paraffin-embedded archival tissue sections from 47 IMTs in 40 patients were immunostained with monoclonal antibodies against ALK and p80. Fluorescence in situ hybridization for ALK rearrangements was done on 22 IMTs from 19 patients. Findings were correlated with clinical features and outcome. RESULTS: ALK positivity was observed in 17 of 47 IMTs (36%) and p80 positivity in 16 of 47 IMTs (34%). Fluorescence in situ hybridization showed ALK rearrangements in nine cases (47%), aneuploidy in three cases (16%), and no rearrangement in seven cases (37%). IMTs with ALK abnormalities by immunohistochemistry and/or fluorescence in situ hybridization originated in the abdomen/pelvis/retroperitoneum, chest, and extremities. The mean age was 6.6 years, with a male/female ratio of 1.3. 64% of patients had no evidence of disease at last follow-up, 45% had one or more recurrences, and 18% displayed histologic evidence of malignant transformation. The IMTs without ALK abnormalities occurred in older children, were more frequent in females, and had fewer recurrences. However, in this group of 40 patients, the differences between the groups with and without ALK abnormalities did not have statistical significance. Aneuploidy without ALK abnormalities was associated with malignant transformation in three of five cases. CONCLUSIONS: Abnormalities of ALK and p80 and evidence of chromosomal rearrangements of 2p23 occur in a significant proportion of IMTs. These changes are most frequent in abdominal and pulmonary IMTs in the first decade of life and are associated with a higher frequency of recurrence. These findings confirm the neoplastic nature of a subset IMT with ALK abnormalities and suggest that aneuploid IMT is a subset with more aggressive clinical behavior. PMID- 11406659 TI - Livers from patients with apolipoprotein A-I amyloidosis are not suitable as "domino" donors. AB - Orthotopic liver transplantation, by eliminating the major site of amyloidogenic protein synthesis, is currently the only definitive treatment of most hereditary amyloidoses. Because of the minimal parenchymal involvement, the explanted livers from familial amyloidotic polyneuropathy (FAP) patients have been transplanted into non-FAP patients in a "domino" fashion. The aim of this study was to evaluate the extent of amyloid deposits in explanted livers from two patients with apolipoprotein A-I amyloidosis, with the Arg26 mutation, to determine their suitability as domino donors. A detailed histologic review of the explanted livers from two patients was performed and assessed for the extent of amyloid deposition by routine and Congo red stains. Both patients had identical histopathologic features. The liver parenchymal involvement was strikingly severe. Large patches of amyloid separated hepatic cords, with accentuation around the central veins. All portal triads were consistently and markedly involved with amorphous eosinophilic deposits within the connective tissue compressing the bile ducts and vascular structures. Hilar vessels had patchy deposits. No involvement of hilar nerve branches was seen. The hepatic parenchyma is extensively involved in hereditary Apolipoprotein A-I amyloidosis, with the Arg26 mutation. These livers, removed at orthotopic liver transplantation, are not suitable for domino donation. PMID- 11406660 TI - Immunoexpression of ultraviolet photoproducts and p53 mutation analysis in atypical fibroxanthoma and superficial malignant fibrous histiocytoma. AB - p53 mutation is one of the major results of ultraviolet (UV) radiation. UV photoproducts of cyclobutane pyrimidine dimers (CPDs) and pyrimidine-pyrimidone (6-4) photoproducts (64PPs) also play an important role in skin cancer development. Atypical fibroxanthoma (AFX), which mimics malignant fibrous histiocytoma (MFH) histologically, occurs in the sun-exposed skin of the elderly, and therefore, an association with UV has long been suspected. Eighteen fibrohistiocytic skin lesions comprising AFX (n = 7), storiform-pleomorphic type MFH centered in the subcutis (superficial MFH; S-MFH; n = 4) and benign fibrous histiocytoma (BFH; n = 7) were used for immunohistochemical and molecular analysis. Eight cases of deep MFH (D-MFH) were also analyzed for UV photoproduct expression for the purposes of comparison. Immunohistochemically, the CPD scores of AFX (3.6 +/- 0.4) were significantly higher than those of S-MFH (1.3 +/- 0.8), D-MFH (0.8 +/- 0.5), or BHF (1.4 +/- 0.7); however, the 64PP scores were extremely low in all these tumors (AFX, 0.1 +/- 0.1; S-MFH, 0.0 +/- 0.0; D-MFH, 0.0 +/- 0.0; and BHF, 0.0 +/- 0.0). AFX, S-MFH, and BFH showed immunoexpression for p53 (2/7, 2/4, and 0/7), respectively. p53 mutations were detected in AFX (4/6; 67%) and S-MFH (1/4; 25%), but not in BFH (0/5; 0%) using polymerase chain reaction-single-strand conformation polymorphism, and all of the mutations in AFX were either C-T transitions or at dipyrimidine sites. In conclusion, AFX and S MFH are both similar fibrohistocytic lesions; however, AFX has high immunoreactivity for CPDs compared with S-MFH, D-MFH, or BFH. These data suggest that CPDs may play an important role in the pathogenesis of AFX. PMID- 11406661 TI - Hematopoiesis/erythropoiesis in myocardial infarcts. AB - Extramedullary hematopoiesis occurring in the myocardium has previously only been reported in a single case of a neonate with cyanotic congenital heart disease. Herein we report the incidental discovery of extramedullary hematopoiesis or pure erythropoiesis in four failing adult hearts with myocardial infarction. In two cases, extramedullary hematopoiesis or erythropoiesis was identified in cardiectomy specimens removed at orthotopic heart transplantation; in two other cases, erythropoiesis was found in left ventricular tissue removed at the time of implantation of left ventricular assist devices. Myocardial hematopoiesis/erythropoiesis was identified based on characteristic light microscopic findings in routinely processed tissue and was confirmed by immunhistochemistry using monoclonal antibodies to the erythroid cell marker glycophorin A (positive in all cases), the megakaryocyte marker CD61, and the granulocyte marker neutrophil elastase (the latter two markers positive in one case only). None of the four patients had a myeloproliferative disorder or evidence of a myelophthisic process. No hematopoietic elements were identified in 109 cardiectomy specimens without acute or recent infarcts. Myocardial hematopoiesis or erythropoiesis could represent heretofore-unrecognized manifestations of altered cytokine expression in patients with heart failure due to myocardial infarction. PMID- 11406662 TI - Pleomorphic rhabdomyosarcoma in adults: a clinicopathologic study of 38 cases with emphasis on morphologic variants and recent skeletal muscle-specific markers. AB - Pleomorphic rhabdomyosarcoma (PRMS) is a rare and controversial tumor of skeletal muscle phenotype. Diagnostic criteria for PRMS by combined histology and currently available immunohistochemistry have not been clearly defined. We report 38 pleomorphic rhabdomyosarcomas in adults, explore morphologic variants, and discuss our experience with both specific and nonspecific skeletal muscle markers in these tumors. Clinical data, morphology, and immunohistochemistry were reviewed. Electron microscopy was performed. Of 38 cases, there were 28 males and 10 females. Patient ages ranged from 21 to 81 years (median = 54 y; mean = 51 y). Tumors were located in the lower extremity (n = 18), abdomen/retroperitoneum (n = 6), chest/abdominal wall (n = 5), spermatic cord/testes (n = 4), upper extremity (n = 3), and one each in the mouth and orbit. Tumor sizes ranged from 1.5 to 15.0 cm (mean = 7.3 cm; median = 6.8 cm). The cases were divided into three variants, each with large, atypical, pleomorphic polygonal rhabdomyoblasts (PRMB) with abundant eosinophilic cytoplasm in varying numbers and different morphologic backgrounds of round or spindled rhabdomyoblasts (RMB). 1. Classic PRMS: Predominantly atypical PRMB in sheets (n = 8). 2. Round cell PRMS: Clusters of PRMB throughout the tumor with a background of slightly atypical, medium-sized, round, blue RMB (n = 13). 3. Spindle cell PRMS: Scattered PRMB in a predominance of atypical spindled RMB arranged in a storiform growth pattern (n = 17). Immunohistochemistry revealed the following: myoglobin (37/38), MyoD1 (19/36), skeletal muscle myogenin (myf4; 19/34), fast skeletal muscle myosin (4/5), desmin (36/38), muscle-specific actin (MSA; 25/35), smooth muscle actin (SMA; 15/33), and muscle specific myogenin (myf3; 25/35). Immunohistochemistry was supportive of skeletal muscle differentiation with at least one positive skeletal muscle specific marker (myoglobin, MyoD1, fast skeletal muscle myosin, or myf4). In addition, all cases had some positivity for nonspecific muscle markers (desmin, MSA, SMA, myf3). Electron microscopy (EM), performed on eight selected cases from all three morphologic groups, demonstrated definitive skeletal muscle differentiation in all cases. Follow-up, available on 30 (79%) cases, revealed that 70% of patients died of disease (mean 20 months, range 1 month-108 months), 3% were alive with disease at 12 months (n = 1); and 27% had no evidence of disease (mean, 83 mo; range, 18 to 108 mo). PRMS, a tumor of predominantly middle aged adult males in the lower extremity, can be diagnosed by the morphologic presence of scattered PRMB with immunohistochemical evidence of at least one skeletal muscle-specific marker. There are three morphologic variants of PRMS. The appropriate diagnosis of PRMS is significant as it is a high-grade sarcoma, with an aggressive clinical course. PMID- 11406663 TI - Sentinel lymph nodes show profound downregulation of antigen-presenting cells of the paracortex: implications for tumor biology and treatment. AB - The sentinel lymph node (SN) is the first node on the direct lymphatic drainage pathway from a tumor. Melanoma-associated SNs are the most likely site of early metastases and their immune functions are strikingly down-modulated. We evaluated histologic and cytologic characteristics of 21 SNs and 21 nonsentinel nodes (NSNs) from melanoma patients who had clinically localized (AJCC Stage I--II) primary cutaneous melanoma. SNs showed highly significant reductions in total paracortical area and in the area of the paracortical subsector occupied by dendritic cells. The frequency of paracortical interdigitating dendritic cells (IDCs) was dramatically reduced in SNs, and most IDCs (approximately 99%) lacked the complex dendrites associated with active antigen presentation. The release of immunosuppressive factors from the primary melanoma may induce a localized and specific paralysis in the SN, which prevents the recognition of otherwise immunogenic melanoma antigens by IDCs. This immune paralysis may facilitate the implantation and growth of melanoma cells in the SN. Cytokine therapy may be able to reverse this immune paralysis. These findings have an important practical application in the histopathologic confirmation that a node is truly sentinel. They also offer an hypothesis to explain the failure of the immune surveillance mechanisms to identify and respond to a small primary melanoma that expresses immunogenic tumor antigens. PMID- 11406664 TI - Multifocal micronodular pneumocyte hyperplasia and lymphangioleiomyomatosis in tuberous sclerosis with a TSC2 gene. AB - A 45-year-old woman with a long-standing diagnosis of tuberous sclerosis (TSC) is presented. She has multifocal micronodular pneumocyte hyperplasia (MMPH) and lymphangioleiomyomatosis (LAM) of the lung, together with the detection of TSC2 gene mutation. During surgery for spontaneous pneumothorax, an open-lung biopsy was performed. Micronodules were well defined, measuring approximately 4 mm in diameter. These MMPHs were histologically composed of papillary proliferation of Type II pneumocytes, with positive immunoreactivity of keratin and surfactant apoprotein. The cystlike spaces, with dilatation and destruction of air spaces, were diffusely formed, and the walls were composed of the spindle cells. Such LAM showed positive immunoreactivity for HMB-45 (a monoclonal antibody specific for human melanoma) and tuberin (the gene product of TSC2). On germline mutation analysis using leukocytes of the present patient, a TSC2 gene mutation was confirmed as a deletion of G (or g) on Exon 9 by polymerase chain reaction-single strand conformational polymorphism. However, no mutation was detected in her son. With microdissection analysis using paraffin-embedding lung tissues, LOH of the TSC2 gene preliminarily was detected in a LAM lesion but not in MMPH. It is suggested that MMPH, in addition to LAM, could be another pulmonary lesion in TSC patients and that the detection of TSC2 and/or TSC1 gene could essentially be useful for the pathogenesis of MMPH and LAM in TSC patients. PMID- 11406665 TI - Primary extrapulmonary sugar tumor (PEST): a report of four cases. AB - The cell of origin and direction of differentiation of the clear cell tumor of the lung (the so-called sugar tumor) remains enigmatic. Recognition of HMB-45 immunoreactivity and identification of melanosomes have suggested a relationship to angiomyolipoma of kidney or liver and lymphangiomyoma. This has given rise to the concept that clear cell tumors are neoplasms of so-called perivascular epithelioid cells--PEComas. Herein we report the existence of four similar tumors occurring in extrapulmonary sites, one of which had malignant features. The three benign tumors occurred in females ages 9, 20, and 40 years; two were located in the rectum and one in the vulva. The malignant tumor occurred in the inter-atrial cardiac septum of a 29-year-old man. Common histologic features were a richly vascular organoid architecture, tumor cells with clear to pale eosinophilic cytoplasm, abundant glycogen, and immunoreactivity for HMB 45, but not S100, multiple keratin, neuroendocrine, or muscle markers. Benign tumors demonstrated low mitotic activity, no necrosis, and good circumscription; the malignant tumor showed considerable mitotic activity, necrosis, and an infiltrative growth pattern. Ultrastructurally, glycogen was present, mitochondria were abundant, and membrane-bound lamellated bodies consistent with premelanosomes were present in two cases, and equivocal in one. Because these tumors have light microscopic, immunohistochemical, and electron microscopic features similar to pulmonary sugar tumors, we propose the name primary extrapulmonary sugar tumor (PEST) for them. Although most PEST's are probably benign, malignant forms appear to exist. These cases further support the concept of a family of systemic HMB-45 positive tumors that include sugar tumors, angiomyolipoma of kidney or liver, lymphangiomyomas, and clear-cell myomelanocytic tumors of the falciform ligament/ligamentum teres. PMID- 11406666 TI - Collecting-duct carcinoma of the kidney with prominent signet ring cell features. AB - We report a case in a 74-year-old woman of collecting-duct carcinoma of the kidney with prominent signet ring cell features. Grossly, the tumor measured 5.5 cm in greatest dimension, occupied the entire upper pole of the kidney, and was well circumscribed. Microscopically, it displayed a predominant tubulopapillary pattern of growth with a hyalinizing stroma. The tumor tubules were lined by a single layer of cells with large, pleomorphic nuclei, some of which had a hobnail appearance. Large intracytoplasmic vacuoles with compression of nuclei (signet ring cells) were present throughout the tumor. Alcian blue, mucicarmine, and periodic acid-Schiff stains failed to identify intracellular mucin or glycogen in the signet ring cells. Enlarged cells with intracytoplasmic vacuoles were also noted in the adjacent collecting ducts. The tumor cells were immunohistochemically positive for cytokeratin including cytokeratin 7, CAM 5.2, AE1/3, and 34 beta E12, vimentin, peanut lectin agglutinin, and Ulex europaeus agglutinin. Electron microscopy revealed that the intracytoplasmic vacuoles were due to intracellular edema. To the best of our knowledge, this is the first reported case of renal collecting-duct carcinoma with prominent signet ring cell features. PMID- 11406667 TI - ADASP recommendations for processing and reporting of lymph node specimens submitted for evaluation of metastatic disease. PMID- 11406669 TI - Evaluation of the accuracy of leukocyte esterase testing to detect pyuria in young febrile children: prospective study. AB - OBJECTIVE: To study the accuracy and clinical application of the dipstick leukocyte esterase test in the detection of pyuria in young febrile children suspected to have urinary tract infection. DESIGN: Prospective study. SETTING: Regional hospital, Hong Kong. PATIENTS: Urine samples were taken from 215 children younger than 2 years who were suspected to have urinary tract infection (fever without an obvious focus of infection). MAIN OUTCOME MEASURES: The accuracy of the dipstick leukocyte esterase test in detecting significant pyuria defined as a leukocyte count >or=10 mm(3) (>or=0.01 x 10(9) /L). RESULTS: Two hundred and fifty-four urine samples collected by bag, midstream clean-catch, suprapubic bladder aspiration, or urethral catheterization were examined. Using urine microscopy results as a reference, the sensitivity and specificity of the leukocyte esterase test in detecting significant pyuria were found to be 72.0% and 85.8%, respectively; the positive and negative predictive values were 55.4% and 92.6%, respectively; and the positive and negative likelihood ratios were 5.1 and 0.3, respectively. CONCLUSIONS: The dipstick leukocyte esterase test cannot accurately detect pyuria in young febrile children. It is also not appropriate as a screening test to exclude pyuria, reduce the need for the microscopic examination of urine, or indicate when a hospital admission for probable urinary tract infection is needed. PMID- 11406670 TI - Rearrangement of the myeloid-lymphoid leukaemia gene in Hong Kong Chinese children with acute leukaemia. AB - OBJECTIVE: To determine the frequency of rearrangement of the myeloid-lymphoid leukaemia gene in acute leukaemia in Hong Kong Chinese children. DESIGN: Immunophenotyping, cytogenetic, and molecular analysis. SETTING: Regional hospital, Hong Kong. PARTICIPANTS: Bone marrow or peripheral blood samples were taken from 27 children aged 16 years or younger with acute leukaemia, from September 1995 through February 1998. MAIN OUTCOME MEASURES: Gene rearrangement was analysed by Southern blotting of HindIII digestion products of mononuclear cell DNA, followed by hybridisation with the myeloid-lymphoid leukaemia P/S4 probe. Nested reverse transcription-polymerase chain reaction analysis was performed to detect and characterise duplication of the myeloid-lymphoid leukaemia gene. RESULTS: Only one (4%) of 23 children whose marrow or peripheral blood samples contained adequate material for genetic study showed rearrangement in the myeloid-lymphoid leukaemia gene. No children were positive for partial tandem duplication of the myeloid-lymphoid leukaemia gene. CONCLUSION: Myeloid lymphoid leukaemia gene rearrangement is rare in Hong Kong Chinese children with acute leukaemia. PMID- 11406671 TI - Valproic acid and thrombocytopenia: cross-sectional study. AB - OBJECTIVE: To investigate the relationship between platelet count and serum valproic acid level, age, duration of valproic acid therapy, and polytherapy, and to determine the clinical significance of thrombocytopenia associated with high dosage valproic acid therapy. DESIGN: Cross-sectional study. SETTING: Residential unit for neurologically impaired children and paediatric out-patient clinic, Hong Kong. PATIENTS: Ninety-six neurologically impaired children who were treated with valproic acid between 1 July 1991 to 3 June 1999. The comparison group consisted of 48 children receiving antiepileptic drugs other than valproic acid. INTERVENTION: Low- or high-dosage valproic acid, using the threshold value of 40 mg/kg/d. MAIN OUTCOME MEASURES: Platelet count and liver function, duration of valproic acid treatment, dosage, and trough serum valproic acid concentration. RESULTS: Seventeen (17.7%) patients in the treatment group developed thrombocytopenia, compared with two (4.2%) in the comparison group (P<0.05). The platelet count was negatively correlated to serum valproic acid level and age, and positively correlated to polytherapy. The duration of valproic acid treatment was not a confounding factor in the age-related decrease in platelet count. Children with a trough level of >450 micromol/L or a daily dose of >40 mg/kg were more likely to develop thrombocytopenia. Thrombocytopenia was mild in most cases. CONCLUSIONS: A trough valproic acid level of >450 micromol/L or a daily dose of >40 mg/kg should alert the clinician to the risk of developing thrombocytopenia. The risk is further increased for older children. The platelet count should be monitored for patients receiving a high concentration of valproic acid who are also receiving drugs that would affect homeostasis, or who are undergoing surgical procedures. PMID- 11406672 TI - Special features of non-melanoma skin cancer in Hong Kong Chinese patients: 10 year retrospective study. AB - OBJECTIVE: To determine the incidence and clinical characteristics of non melanoma skin cancer in Hong Kong Chinese patients. DESIGN: Retrospective study. SETTING: Social Hygiene Services, Hong Kong. PATIENTS: Records of 528 Chinese patients with a histological diagnosis of non-melanoma skin cancer from 1990 to 1999 were reviewed. MAIN OUTCOME MEASURES: Demographic data, site and clinical type of cancer, predisposing factors, history, recurrence, and the development of new skin cancers. RESULTS: Non-melanoma skin cancer is uncommon but not rare among the Chinese population in Hong Kong. The incidence of newly diagnosed basal cell carcinoma in 1990 was 16.0 per 10,000 new skin case attendances and, in 1999, the incidence was 31.8 per 10,000 new skin case attendances. The corresponding figures for squamous cell carcinoma in 1990 and 1999 were 6.9 and 11.6 per 10,000 new skin case attendances. The incidence of basal cell carcinoma among the Hong Kong Chinese population in 1990 and 1999 was 0.32 and 0.92 per 100,000, respectively, whereas that of squamous cell carcinoma was 0.16 and 0.34 per 100,000, respectively. Demographic data and the site distribution of non melanoma skin cancer were comparable to those reported in Caucasians living in North America and Europe, but different from those in Caucasians living in Australia and Hawaii. Pigmented basal cell carcinoma was the most common type of non-melanoma skin cancer (60.1%) in Chinese patients, in contrast with rodent ulceration in Caucasian. Multiple skin cancers, recurrence, and subsequent new skin cancers were less frequently observed than in studies of Caucasians. CONCLUSION: When compared with reported findings in Caucasians, Chinese patients show differences in the clinical type and multiplicity of lesions, predisposing factors, recurrence, and subsequent new skin cancer rates for non-melanoma skin cancer. Pigmented basal cell carcinoma seems to be an important differential diagnosis with regard to pigmented lesions in the Chinese population. PMID- 11406673 TI - Epidemiology of headache in Hong Kong primary-level schoolchildren: questionnaire study. AB - OBJECTIVE: To collect and analyse epidemiological data on childhood headache in the Hong Kong community. DESIGN: Questionnaire study. SETTING: Three primary schools, Hong Kong. PARTICIPANTS: Two thousand, one hundred and twenty pupils from 2156 replied to the questionnaire survey. One hundred and twenty-four pupils who were identified to have suspected recurrent headache were invited to a follow up medical consultation. MAIN OUTCOME MEASURES: Age-groups and prevalence of tension-type and migraine headache, using the diagnostic criteria of the International Headache Society. RESULTS: The overall prevalence of headache in the 2120 respondents was 2.8%. The prevalence of tension-type headache, migraine, probable migraine, and unclassified headache were 1.2%, 0.5%, 0.7%, and 0.5%, respectively. The age-specific prevalence of headache from the age of 6 to 13 years showed a steadily increasing trend from childhood to the early teens. CONCLUSION: Headache is a common complaint for children, although it may be underrecognised. Further study will be beneficial for providing better management of headache in this population. PMID- 11406674 TI - Surgical management of Parkinson's disease: a critical review. AB - Parkinson's disease is a progressive disabling movement disorder that is characterised by three cardinal symptoms: resting tremor, rigidity, and bradykinesia. Before the availability of effective medical treatment with levodopa and stereotactic neurosurgery, the objective of surgical management was to alleviate symptoms such as tremor at the expense of motor deficits. Levodopa was the first effective medical treatment for Parkinson's disease, and surgical treatment such as stereotactic thalamotomy became obsolete. After one decade of levodopa therapy, however, drug-induced dyskinesia had become a source of additional disability not amenable to medical treatment. Renewed interest in stereotactic functional neurosurgery to manage Parkinson's disease has been seen since the 1980s. Local experience of deep-brain stimulation is presented and discussed in this paper. Deep-brain stimulation of the subthalamic nucleus is an effective treatment for advanced Parkinson's disease, although evidence from randomised control trials is lacking. PMID- 11406675 TI - The impact of molecular biology on clinical neurology. AB - Advances in molecular biology have increased our understanding of both inherited and sporadic forms of neurological disease. In this review, the impact of these advances is discussed in relation to specific neurological conditions. These include the hereditary neuropathies and ataxias, Huntington's disease, and the muscular dystrophies, as well as Alzheimer's disease, Parkinson's disease, and motor neuron disease. Genetic channelopathies, such as familial hemiplegic migraine, are also described. Although knowledge in this area overall is still relatively scant, current advances in molecular biology have helped in the reclassification of some neurological disorders, thereby providing a further step towards the development of rational therapies to treat these conditions. PMID- 11406676 TI - Cerebrovascular disease among Chinese populations--recent epidemiological and neuroimaging studies. AB - Stroke mortality in individuals of Chinese descent has declined during the past decades, although the absolute number of patients with first-ever stroke is escalating. This review summarises recent epidemiological and imaging studies conducted among Chinese populations. Data indicating differences between Chinese and Caucasian ethnic groups in the pathophysiological mechanisms of stroke are highlighted, and the potential implications of these findings for prevention and management of stroke are discussed. PMID- 11406677 TI - Cerebrovascular disease--advances in management. AB - Recent advances in the diagnosis and treatment of stroke have justified its management as a medical emergency. This article summarises the current recommendations for the initial management of major types of stroke with emphasis on acute therapy for ischaemic stroke. Recommendations are based on results of well-designed clinical trials. An acute stroke care team and an acute stroke unit should be established in all regional hospitals. Diagnosis of stroke must be accurate. General management aims for prevention and treatment of neurological and systemic complications, whereas specific management varies according to the stroke type and the underlying pathogenic mechanisms. For selected patients with ischaemic stroke, intravenous recombinant tissue plasminogen activator or a modified viper venom within 3 hours of onset, or intra-arterial pro-urokinase within 6 hours may improve functional outcomes. Neurosurgical treatment is indicated for some patients with ischaemic or haemorrhagic strokes. Prevention of recurrence and rehabilitation are the core components of subsequent management. PMID- 11406678 TI - Infections of the nervous system: an update on recent developments. AB - The past decade has seen major changes in the field of infectious diseases. In particular, many new infections of the nervous system have been recognised, including the lethal infections of Enterovirus 71, and the Nipah and West Nile viruses. Increased interest in prion diseases has occurred, following the recognition of animal-to-human transmission in Europe. Familiar bacteria such as the pneumococcus continue to cause problems due to increasing resistance to multiple antibiotics. Furthermore, human immunodeficiency virus-infected and other immunocompromised patients are under the constant threat of opportunistic infections, many of which are targeted towards the brain and spinal cord. This paper reviews the changing world of nervous system infections, highlighting some of the most significant recent developments. PMID- 11406679 TI - Recent advances in the diagnosis and management of epilepsy. AB - Epilepsy is the second most common neurological disorder in developed countries. Recent advances in the understanding of pathogenetic mechanisms and in the diagnosis and management of epilepsy over the past decade have had a significant impact on every aspect of epilepsy management. A simple diagnosis is inadequate for both patients and physicians. A full diagnostic investigation to classify the type of epilepsy is now mandatory for appropriate care. In this review, current views on pathogenetic mechanisms, controversies in classification, advances in neuroimaging techniques, the use of novel antiepileptic drugs, and non pharmacological treatment options for intractable epilepsy will be discussed. PMID- 11406680 TI - Percutaneous balloon mitral valvuloplasty during pregnancy: long-term follow-up of infant growth and development. AB - We have studied the long-term growth and development of seven infants who had been exposed during the foetal stage to ionising irradiation associated with percutaneous balloon mitral valvuloplasty between 1 April 1991 and 30 November 1993. We found that infants of mothers who underwent this procedure during the second and third trimester of pregnancy had normal long-term growth and development. This finding supports the argument that percutaneous balloon mitral valvuloplasty using the Inoue balloon technique should be the procedure of choice for pregnant women who have severe mitral stenosis refractory to medical therapy. PMID- 11406681 TI - Meningoencephalitis caused by Bacillus cereus in a neonate. AB - We report on a newborn boy, who was delivered at 26 weeks' gestation by emergency caesarean section because of a prolapsed cord and breech presentation. Grade IV hyaline membrane disease subsequently developed, for which a surfactant was given. On day 8, there were frequent apnoeic attacks, and on day 30, marked irritability developed, as did intermittent stiffening of all four limbs. The anterior fontanelle was bulging and tense, and the cerebrospinal fluid was found to be turbid. Gram staining of the cerebrospinal fluid and blood revealed Gram positive bacilli. Subsequent culturing yielded Bacillus cereus, which was sensitive to amikacin and vancomycin. Severe cerebral oedema developed, however, and computed tomography of the brain showed bright cortical sulci, suggestive of meningitis. The baby died on day 37, and post-mortem histological examination of the brain showed extensive liquefactive necrosis with abundant neutrophilic infiltration. Since infection with Bacillus cereus is rapidly fatal, early recognition of infection by this organism is important. PMID- 11406682 TI - Classic late infantile neuronal ceroid lipofuscinosis in a Chinese patient. AB - Neuronal ceroid lipofuscinoses are a group of rare neurodegenerative disorders that are characterised by an accumulation of autofluorescent lipopigments in neurons and extraneuronal tissues. We report on a 4-year-old boy who presented with an acute onset of seizures followed by rapid psychomotor deterioration, ataxia, and visual failure. Photic stimulation at 1 to 3 Hz elicited discrete spike and wave discharges in the electroencephalogram, which were diminished at a higher frequency of stimulation. The electroretinogram was extinct. Magnetic resonance imaging of the brain showed generalised cerebral and cerebellar atrophy. Electron microscopic examination of lymphocytes and samples of muscle and skin revealed characteristic curvilinear inclusion bodies. To our knowledge, this is the first case of late infantile neuronal ceroid lipofuscinosis to be reported in a Hong Kong Chinese patient. PMID- 11406683 TI - Surgery for intractable epilepsy in a 14-year-old girl. AB - We report on a 14-year-old girl who presented with a 2-year history of simple and complex partial seizures with secondary generalisation. Monotherapy using carbamazepine and combination therapy using carbamazepine and gabapentin had been tried within the 2 years before presentation. Seizure control, however, was poor. Magnetic resonance imaging showed structural abnormalities over the right occipital and temporal lobes. Continuous scalp video electroencephalography was performed over 4 days, during which six clinical seizures were associated with electroencephalography changes at the right occipital and temporal lobes. Invasive intracranial video electroencephalography identified a focus at the right occipital lobe, a focus at the right temporal lobe that spread rapidly to the right parietal lobe, and an irritative zone over the posterior part of the right frontal lobe. Functional mapping delineated the motor and sensory cortices. Right temporal lobectomy, right occipitoparietal cortical excision, and multiple subpial transections of the posterior part of the right frontal lobe were performed. For 16 months after the surgery, the patient has been seizure-free while receiving drug treatment, and the only complication reported has been a segmental loss of the left visual field. PMID- 11406684 TI - Neuroleptic malignant syndrome induced by droperidol. AB - A case of droperidol-induced neuroleptic malignant syndrome during anaesthesia is presented. An 86-year-old man underwent spinal anaesthesia for open reduction and internal fixation of a trochanteric hip fracture. He received droperidol 5 mg intravenously for sedation towards the end of surgery. He subsequently became very drowsy and experienced marked muscle rigidity and autonomic instability. He became febrile postoperatively. The clinical syndrome resolved after 12 hours. When using droperidol in anaesthesia or intensive care--especially when large doses are given--the development of neuroleptic malignant syndrome should be suspected if the patient becomes febrile and has muscle rigidity and autonomic instability. PMID- 11406685 TI - Effects of Experience and Environment on the Developing and Mature Brain: Implications for Laboratory Animal Housing. AB - Decades of research have determined that an animal's brain structure and behavior are molded by experience. Expected experience that plays a critical role in early organization of the brain may be encoded via a process of overproduction of synaptic connections followed by the loss of those that are underutilized during a critical period. However, novel information may be encoded throughout life by the formation of new synapses as the individual animal is exposed to new environmental stimuli. Many laboratory species reared in complex environments or trained to perform complex tasks, regardless of the age when the altered experience is introduced, will exhibit an increase in the number of synapses per neuron as well as other anatomical differences from those reared in standard laboratory housing. Nevertheless, even though increased environmental stimulation may result in more "normal" anatomical and physiological development for that species, there is no conclusive evidence that enriched caging is essential or even that it increases well-being in laboratory rodents. PMID- 11406686 TI - Floor Space Needs for Laboratory Mice: C56BL/6 Males in Solid-bottom Cages with Bedding. AB - Measures of performance, mortality, adrenal weights, plasma glucocorticoid concentration, and selected immune measures were collected in an attempt to define space needs of laboratory mice. Six replications of 3 C57BL/6 male mice per cage were examined while housed on bedding at 5, 10, 15, or 20 in(2) (32.2, 64.5, 96.8, or 129 cm(2)) per mouse. Body weights were not influenced by treatment; however, mice in smaller spaces (5 in(2) per mouse) consumed or wasted more feed and water than mice given greater space allowances. Mice given the least amount of space (5 in(2) per mouse) had greater lymphocyte proliferation in response to the T-cell mitogen PHA than mice given more space. Mice provided 10 in(2) per mouse had greater natural killer cytotoxicity than mice given greater or less space. Mouse mortality was greater as more space was provided. In contrast, adrenal weights and plasma glucocorticoid concentrations were progressively greater with lower space allowances. The National Research Council 1996 recommendation of 15 in(2) per mouse, for this strain and sex of mice, would result in greater mortality and reduced activity of some immune measures. Socially housed male C57BL/6 mice will benefit from less space than recommended by the National Research Council in 1996. PMID- 11406687 TI - Opportunistic Infections: Why Worry? AB - The proliferation and increased use of genetically manipulated rodents are taxing laboratory animal facilities and diagnostic laboratories. Although serological testing remains the standard for monitoring immunologically intact animals, molecular diagnostics are playing increasing roles in detecting infections in animals that have compromised immune systems. The importance of these assays is particularly evident now that we recognize that genetic manipulation can result in unanticipated phenotypes, including ones with decreased resistance to infectious diseases. PMID- 11406688 TI - Deer Mice As Laboratory Animals. AB - Although laboratory mice (Mus) and rats (Rattus) are the most widely used research rodents, deer mice (Peromyscus maniculatus) and their congeneric species are favored as nontraditional alternatives for some purposes. Mice of the native genus Peromyscus are the most abundant and widely distributed rodents in North America. They occur in a great diversity of habitats and play a significant role in natural ecosystems. Because of their abundance, peromyscines are commonly hosts for larva of ticks that transmit Lyme disease bacteria, and they are implicated in several other vector-borne diseases. Deer mice also are the principal carriers of the virus that causes hantaviral pulmonary syndrome, or "Four Corners disease." Deer mice are useful as laboratory models for a variety of other types of pure and applied research. They are easily maintained and bred in captivity using the husbandry protocols developed for other small laboratory rodent species. The Peromyscus Genetic Stock Center at the University of South Carolina maintains more than 50 laboratory-bred, well-characterized stocks of deer mice and other peromyscine species for research and educational use. PMID- 11406689 TI - Inflammatory Models of Pain and Hyperalgesia. AB - This article addresses important pain research models in nonhuman animals. These models attempt to mimic human persistent pain conditions. Models of persistent pain employ inflammatory agents that produce discomfort and hyperalgesia (i.e., an enhanced response to a noxious stimulus). The models are associated with skin, subcutaneous tissue, and joint inflammation (somatic structures). Studies employing such models have led to significantly improved understanding of mechanisms of somatic pain. It is important that investigators assess the level of pain produced in these animals and provide analgesic agents whenever it does not interfere with the purpose of the experiment. Pain can be inferred from ongoing behavioral variables such as feeding and drinking, sleep-waking cycle, grooming, and social behavior. PMID- 11406690 TI - Models of Visceral Nociception. AB - Pains arising from the viscera constitute a large portion of clinically treated pains. They are characterized by poor localization; immobility with tonic increases in muscle tone; and vigorous but nonspecific changes in autonomic function, such as changes in respiration, heart rate, and blood pressure. Tissue damaging stimuli do not reliably produce visceral pain, so the study of visceral nociception in nonhuman animals requires identification of appropriate stimuli and responses. This article defines "noxious" visceral stimuli as those that produce pain in humans, result in aversive behaviors in animals, and evoke responses that are inhibited by manipulations known to be analgesic in humans. To be valid, the measured responses must be reliable, inhibited by known analgesics, and not inhibited by nonanalgesics. Using these criteria as measures of validity, the author examined several visceral pain models. The writhing test (application of intraperitoneal irritants) failed to meet these criteria; however, responses to small bowel distension, colonic-rectal distension, artificial ureteral calculosis, urinary tract distension, and the intravesical application of irritants met most, if not all, of the criteria. Other models, such as responses to biliary system distension, to reproductive organ stimulation, to the focal application of algesic agents onto various viscera, and to ischemic stimuli, met some of these criteria. This information should assist readers in decisions related to the use of visceral pain models. PMID- 11406691 TI - Postoperative Models of Nociception. AB - Even though it is a costly, poorly understood problem, very little effort has been dedicated toward research on the mechanisms of acute postoperative pain. Presumably, if we learn more about the etiology of acute incisional pain and the sensory processes that intensify pain after surgery, new treatment methods can be advanced. Since the mid-1990s, our group and others have developed and characterized models for postoperative pain. In one model, a hind paw plantar incision is made. Persistent reduced withdrawal thresholds to mechanical stimulis suggesting hyperalgesia are present. No tonic or spontaneous pain is apparent, but diminished weight bearing is noted on the incised hind paw. Pain-related behaviors remain remarkable for several days and then gradually decrease. The model therefore has similarities to the time course for pain in postoperative patients. Ovariohysterectomy, a clinically relevant procedure, has been used to study postoperative pain. Both rat and dog models have been studied, and a variety of pain-related behaviors including pain at rest and wound sensitivity have been examined. These models will improve our understanding of unique pain mechanisms caused by particular injuries. As we understand postoperative pain and determine the unique mechanisms for acute tissue injury pain, better treatments will evolve and perioperative morbidity will decrease. PMID- 11406692 TI - New Frontiers in Cytokine Involvement during Experimental Sepsis. AB - Despite significant advances in the antibiotic arsenal and in intensive care unit technology, including mechanical ventilation, sepsis-related morbidity and mortality remain unacceptably high. Ultimately, 25 to 50% of all septic episodes end in death. However, various subsets of septic patients, including those who experience septic peritonitis, and various secondary sequelae like the acute respiratory distress syndrome or nosocomial infections, demonstrate much higher mortality rates ranging from 60 to 95%. Although a number of strategies have been utilized to curb the progression of systemic inflammatory response syndrome with immune or inflammatory modulating therapies, none of these interventions has resulted in significant improvement in survival, and some have proven deleterious. The inability to utilize immune-modulating strategies effectively to treat septic patients likely reflects the inherent conflict that is illustrated by the two diagnostic criteria for the syndrome. The very immune/inflammatory response that has evolved to eliminate infection results in severe and life threatening damage to host tissues. This review outlines the inflammatory pathways utilized by the host during a septic response. The basis of early immune modulating therapies and possible reasons these approaches have failed in the treatment of sepsis are discussed. A picture of the ideal therapeutic approach for acute inflammatory diseases like sepsis is also created, and the reason therapies targeting chemokine pathways may more closely approximate the ideal therapy is proposed. PMID- 11406693 TI - Understanding the Pathogenesis of Inflammation Using Rodent Models: Identification of a Transcription Factor (NFkappaB) Necessary for Development of Inflammatory Injury. AB - The acute inflammatory response is composed of a complex cascade of mediators, which facilitate the removal of infectious agents and restore normal tissue function. Experiments using cultured cells have demonstrated that the transcription factor nuclear factor kappa B (NFkappaB) is critically important for the expression of numerous proinflammatory mediators. However, extrapolation of data derived from these types of studies often does not represent the in vivo response accurately. Application of rodent models of inflammation has allowed detailed study of the role of NFkappaB and its transcriptional products in a variety of inflammatory diseases. This article reviews the importance of rodent models for the study of complex biological systems, such as acute inflammation, and presents ways in which these models have been used to characterize the requisite function of NFkappaB in these responses. PMID- 11406694 TI - Cytokine and Chemokine Interactions in Allergic Airway Inflammation. AB - Allergic airway inflammation is characterized by peribronchial eosinophil accumulation within the submucosa surrounding the airway. The development of appropriate animal models to dissect the critical mechanisms involved in the deleterious responses is crucial for the evolution of proper therapeutic approaches. This article reviews several developed models of allergic airway inflammation induced by intratracheal challenge with parasite (Schistosoma mansoni egg antigen [SEA]), environmental (cockroach), and fungal (Aspergillus) allergens in presensitized mice. Use of these models to examine the eosinophil recruitment has identified key cytokines and adhesion molecules involved in the responses, which rely on several interacting events. These cytokines include interleukin 4 (IL-4), IL-13, and a number of chemokines, which appear to act as activating and recruitment PMID- 11406695 TI - Role and Regulation of Chemokines in Rodent Models of Lung Inflammation. AB - Acute inflammatory lung injury has been induced in rats by intrapulmonary deposition of immunoglobulin G immune complexes or instillation of bacterial lipopolysaccaride. Several juxtaposed cysteine residue (CXC) chemokines (e.g., macrophage inflammatory protein 2 [MIP-2] and cytokine-induced neutrophil chemoattractant [CINC]) and CC chemokines (MIP-1alpha, MIP-1Beta, and monocyte chemoattractant protein-1 [MCP-1]) are upregulated in these acute injury models and appear in substantial amounts in bronchoalveolar (BAL) fluids during the inflammatory response. Antibody-induced blockade of either tumor necrosis factor alpha (TNFalpha ) or the complement activation product C5a results in significant reductions in BAL levels of chemokines, causing depressed inflammatory responses and diminished lung injury. These data suggest that both TNFalpha (a product of activated macrophages) and C5a participate as positive feedback mediators, resulting in maximal expression of chemokines. These chemokines are involved in recruitment of neutrophils and activation of tissue macrophages, the collective products of which cause acute lung injury mediated by the generation of oxidants and release of proteases. PMID- 11406696 TI - Use of Immunodeficient Mice for the Evaluation of CXC Chemokines in the Regulation of Tumor-associated Angiogenesis. AB - Angiogenesis, defined as the growth of new capillaries from pre-existing vessels, is a pervasive biological phenomenon that is at the core of many physiologic and pathologic processes such as tumor growth. The use of human tumor xenografts in immunodeficient mice has provided significant insight into the biology of angiogenesis as it relates to tumor growth and metastasis. Work reviewed in this article supports the notion that net tumor-derived angiogenesis during tumorigenesis of human tumors is determined, in part, by an imbalance in favor of the overexpression of angiogenic (compared with angiostatic) juxtaposed cysteine residue (CXC) chemokines. This paradigm predicts an environment that favors angiogenesis (tumorigenesis) and supports the potential for spontaneous metastases. The article describes the use of immunodeficient mice as an animal model system for characterizing the qualitative and quantitative presence of these angiogenic and angiostatic CXC chemokines during tumorigenesis, as well as determining their net contribution to human tumorigenesis and metastasis in vivo. Various cancer cell lines have been used and xenografted into immunodeficient mice to create human tumor/mouse chimeras, indicating that an imbalance in the biology of angiogenic versus angiostatic CXC chemokines supports a significant portion of human tumor-derived angiogensis that leads to augmented tumorigenesis and spontaneous metastases. It has also been possible to identify potentially therapeutic novel strategies to manipulate the imbalance of angiogenic (compared with angiostatic) CXC chemokines, which may be directly translational to human disease. PMID- 11406697 TI - Chemokine Regulation of Immune-mediated Demyelinating Disease. AB - Experimental autoimmune encephalomyelitis (EAE) is a CD4+ Th1-mediated demyelinating disease of the central nervous system (CNS), which serves as a model for multiple schlerosis (MS). A hallmark in the pathogenesis of this disease is the emigration of T cells and monocytes from the blood to the CNS. Chemokines are small-molecular-weight chemotactic peptides, which are ligands for seven transmembrane-spanning, G protein-coupled receptors and which deliver signals leading to a variety of T cell functions including costimulation, cytokine expression, differentiation, and integrin activation. Several considerations suggest a role for chemokines in the influx of inflammatory cells to the CNS and the resulting disease process, including a tight temporal expression pattern with a relation to disease activity and prevention of disease development by in vivo neutralization. This article reviews the evidence that temporal and spatial expressions of chemokines are critical factors that regulate EAE, which makes this an appropriate animal model to study the pathogenesis of MS disease activity. PMID- 11406698 TI - Research techniques for the squirrel monkey (Saimiri sp.). PMID- 11406699 TI - Steroid resistance in the squirrel monkey: an old subject revisited. PMID- 11406700 TI - Defining the moribund condition as an experimental endpoint for animal research. PMID- 11406701 TI - A systematic approach for establishing humane endpoints. PMID- 11406702 TI - Humane endpoints for genetically engineered animal models. PMID- 11406703 TI - Humane endpoints for infectious disease animal models. PMID- 11406704 TI - Humane endpoints and acute toxicity testing. PMID- 11406706 TI - Structures and limits of animal models: examples from alcohol research. PMID- 11406707 TI - Aggression in knockout mice. PMID- 11406708 TI - Developing standardized behavioral tests for knockout and mutant mice. PMID- 11406709 TI - Nonhuman primates: a critical role in current disease research. AB - This review article emphasizes the critical role of nonhuman primates (NHPs) in biomedical research. It focuses on the most recent contributions that NHPs have made to the understanding, treatment, and prevention of important infectious diseases (e.g., acquired immunodeficiency syndrome, hepatitis, malaria) and chronic degenerative disorders of the central nervous system (e.g., Parkinson's and Alzheimer's diseases). The close phylogenetic relation of NHPs to humans not only opens avenues for testing the safety and efficacy of new drugs and vaccines but also offers promise for evaluating the potential of new gene-based treatments for human infectious and genetic diseases. PMID- 11406710 TI - Perspectives on hepatitis B studies with chimpanzees. AB - Chimpanzees have been shown to be exquisitely susceptible to human hepatitis viruses, without themselves developing clinical illness, thus providing an important model for studies on these agents. Chimpanzees have contributed substantially to human welfare by making possible the development of hepatitis B vaccines, which now prevent development of cirrhosis and hepatocellular carcinoma in millions of people. They have provided a means to evaluate the efficacy of virus inactivation strategies, which have made blood derivatives formerly contaminated with blood-borne viruses (hepatitis B, C, and human immunodeficiency viruses) safe with respect to their transmission. In exchange for these contributions, humans owe chimpanzees lifelong retirement in sanctuaries that offer socialization and environmental enrichment. PMID- 11406711 TI - The woodchuck model of hepatitis B virus infection. AB - The woodchuck hepatitis virus (WHV) was the first of the mammalian and avian hepadnaviruses described after discovery of the virus of hepatitis B (HBV). Woodchucks chronically infected with WHV develop progressively severe hepatitis and hepatocellular carcinoma, which present as lesions that are remarkably similar to those associated with HBV infection in humans. The initial virological studies and studies of pathogenesis utilized woodchucks that had been trapped in the wild and had acquired WHV infection naturally. Research with wild woodchucks was complicated by lack of knowledge of their backgrounds (e.g., dietary history, exposure to parasites or environmental toxins, and source and duration of WHV infection). Breeding colonies of woodchucks have been established and maintained in laboratory animal facilities, and laboratory-reared woodchucks are superior for experimental studies of pathogenesis or hepatocarcinogenesis. It is possible to infect neonatal woodchucks born in the laboratory with standardized inocula and produce a high rate of chronic WHV carriers that are useful for controlled investigations. WHV has been shown experimentally to cause hepatocellular carcinoma, supporting conclusions based on epidemiological and molecular virological studies that HBV is an important etiological factor in human hepatocarcinogenesis. Chronic WHV carrier woodchucks have become a valuable animal model for the preclinical evaluation of antiviral therapy for HBV infection, providing useful pharmacokinetic and pharmacodynamic results in a relevant animal disease model. It also has been shown that the pattern of toxicity and hepatic injury observed in woodchucks treated with certain fluorinated pyrimidines is remarkably similar to that observed in humans that were treated with the same drugs, suggesting the woodchuck has significant potential for the preclincial assessment of antiviral drug toxicity. PMID- 11406712 TI - Animal models of hepatitis delta virus infection and disease. AB - Hepatitis delta virus (HDV) is a defective RNA virus with similarities to unusual subviral pathogens of higher plants. It requires hepatitis B virus (HBV) for its replication/transmission, and HBV-infected humans are the only established host. HDV causes both severe acute hepatitis and rapidly progressive chronic disease in some individuals. The HDV life cycle involves remarkable features, such as ribozyme- mediated autocatalytic processes, Pol II-directed RNA synthesis from a single-stranded circular RNA template, and RNA editing. Much of our understanding of the nature of this pathogen derives from experimental studies in the chimpanzee model of HBV infection. The hepadnavirus-infected eastern woodchuck also is capable of supporting HDV replication and offers opportunities for the development of control strategies that might be applicable to human type D hepatitis. PMID- 11406713 TI - Hepatitis C: a brief clinical overview. AB - Hepatitis C has emerged as an important public health problem that has affected 3.9 million Americans and 170 million people worldwide and is currently the most common indication for orthotopic liver transplantation. The disease, characterized by asymptomatic onset, is often discovered incidentally through blood tests obtained during routine physical examination or before blood donation. Spontaneous recovery occurs in about 20% of patients. Among those who remain chronically infected, an equal percentage progress to cirrhosis within 20 yr, have stable nonprogressive disease, or progress more slowly over 40 to 60 yr. At present, combination therapy with interferon plus ribavirin is the treatment of choice for hepatitis C-infected patients identified as appropriate candidates for therapy. Unfortunately, sustained response rates are only modest, with a lesser response among African Americans, and treatment is associated with a number of side effects. Research studies attempting to improve the response to current therapy, to identify alternative treatments or treatment strategies, and to develop an effective vaccine are ongoing and will hopefully provide us with the ability to better understand and manage hepatitis C. PMID- 11406714 TI - The chimpanzee model of hepatitis C virus infections. AB - The chimpanzee (Pan troglodytes) is the only experimental animal susceptible to infection with hepatitis C virus (HCV). The chimpanzee model of HCV infection was instrumental in the initial studies on non-A, non-B hepatitis, including observations on the clinical course of infection, determination of the physical properties of the virus, and eventual cloning of the HCV nucleic acid. This review focuses on more recent aspects of the use of the chimpanzee in HCV research. The chimpanzee model has been critical for the analysis of early events in HCV infection because it represents a population for which samples are available from the time of exposure and all exposed animals are examined. For this reason, the chimpanzee represents a truly nonselected population. In contrast, human cohorts are often selected for disease status or antibody reactivity and typically include individuals that have been infected for decades. The chimpanzee model is essential to an improved understanding of the factors involved in viral clearance, analysis of the immune response to infection, and the development of vaccines. The development of infectious cDNA clones of HCV was dependent on the use of chimpanzees, and they will continue to be needed in the use of reverse genetics to evaluate critical sequences for viral replication. In addition, chimpanzees have been used in conjunction with DNA microarray technology to probe the entire spectrum of changes in liver gene expression during the course of HCV infection. The chimpanzee will continue to provide a critical aspect to the understanding of HCV disease and the development of therapeutic modalities. PMID- 11406715 TI - New animal models of hepatitis B and C. AB - The narrow host range of infection and lack of suitable tissue culture systems for the propagation of hepatitis B and C viruses are limitations that have prevented a more thorough understanding of persistent infection and the pathogenesis of chronic liver disease. With hepatitis B virus (HBV), this lack of knowledge has been partially overcome by the discovery and characterization of HBV-like viruses in wild animals. With hepatitis C virus (HCV), related flaviviruses have been used as surrogate systems for such studies. Other laboratories have developed transgenic mice that express virus gene products and/or support virus replication. Some HBV transgenic mouse models develop fulminant hepatitis, acute hepatitis, or chronic liver disease after adoptive transfer, and others spontaneously develop hepatocellular carcinoma (HCC), as in human infections. Among HCV transgenic mice, most develop no disease, but acute hepatitis has been observed in one model, and HCC in another. Although mice are not susceptible to HBV and HCV, their ability to replicate these viruses and to develop liver diseases characteristic of human infections provides new opportunities to study pathogenesis and develop novel therapeutics. PMID- 11406716 TI - Molecular clones of hepatitis C virus: applications to animal models. AB - Hepatitis C virus (HCV) is a global public health problem, with approximately 3% of the world population now infected. The clinical course of HCV often involves chronic infection, which can lead to liver dysfunction and hepatocellular carcinoma. Because HCV cannot be efficiently propagated in cell culture, researchers have relied heavily on animal models to study the physical characteristics of HCV and the course of events associated with HCV infection. The chimpanzee is the only nonhuman primate actually proven to be susceptible to HCV infection and has commonly been used to study viral hepatitis induced by HCV. Molecular cloning of the HCV genome has now allowed HCV transmission studies in chimpanzees to progress from the early work of characterizing infectious serum to a current focus of characterizing infectious HCV molecular clones. Moreover, the cloned HCV genome has paved the way for the development of alternative animal models for HCV, most notably transgenic mouse models for the study of HCV pathogenesis. The authors review these animal model applications of the HCV molecular clones, including construction and transmission of mutant viral genomes. The expression of specific viral protein products in these animal models will provide important insight into the structure-function relation that specific HCV genome sequences impart on virus replication and pathogenesis. PMID- 11406717 TI - GB virus B as a model for hepatitis C virus. AB - GB viruses A and B (GBV-A and GBV-B) are members of the Flaviviridae family and are isolated from tamarins injected with serum from a human hepatitis patient. Along with a related human virus, GB virus C, or alternatively, hepatitis G virus (GBV-C/HGV), the three viruses represent the GB agents. Of the three viruses, GBV B has been proposed as a potential surrogate model for the study of hepatitis C virus (HCV) infections of humans. GBV-B is phylogenetically most closely related to HCV and causes an acute, self-resolving hepatitis in tamarins as indicated by an increase in alanine aminotransferase and changes in liver histology. Similarities between GBV-B and HCV are found at the nucleotide sequence level with the two viruses sharing 28% amino acid homology over the lengths of their open reading frames. Short regions have even higher levels of homology that are functionally significant as shown by the ability of the GBV-B NS3 protease to cleave recombinant HCV polyprotein substrates. The shared protease substrate specificities suggest that GBV-B may be useful in testing antiviral compounds for activity against HCV. Although there are numerous similarities between GBV-B and HCV, there are important differences in that HCV frequently causes chronic infections in people, whereas GBV-B appears to cause only acute infections. The acute versus chronic course of infection may point to important differences between the two viruses that, along with the numerous similarities, will make GBV B in tamarins a good surrogate model for HCV. PMID- 11406718 TI - Animal models of hepatitis A and E. AB - Several useful animal models for both hepatitis A and E have been identified, characterized, and refined. At present, all of the best models utilize nonhuman primates: chimpanzees, tamarin species, and owl monkeys for hepatitis A; and macaque species, chimpanzees, and owl monkeys for hepatitis E. Pigs may prove useful for some studies of hepatitis E, and it is hoped that serological evidence of widespread infection of rats with an HEV-like agent may lead to the development of an animal model based on laboratory rats. As has been the case for each of the hepatitis viruses as they have been discovered, the development of useful and reproducible animal model systems has been critical for moving the field forward as expeditiously as possible. PMID- 11406719 TI - Nuclear magnetic resonance spectroscopy and imaging in animal research. AB - Nuclear magnetic resonance (NMR) spectroscopy and imaging can be used to investigate, noninvasively, a wide range of biological processes in systems as diverse as protein solutions, single cells, isolated perfused organs, and tissues in vivo. It is also possible to combine different NMR techniques enabling metabolic, anatomical, and physiological information to be obtained in the same experiment. This review provides a simple overview of the basic principles of NMR and outlines both the advantages and disadvantages of NMR spectroscopy and imaging. A few examples of potential applications of NMR spectroscopy and imaging are presented, which demonstrate the range of questions that can be asked using these techniques. The potential impact of using NMR techniques in a biomedical research program on the total number of animals required for specific investigations, as well as the number of animals used in research, are discussed. The article concludes with a personal perspective on the impact of continuing improvements in NMR technology for future applications in animal research. PMID- 11406720 TI - Electron paramagnetic resonance for small animal imaging applications. AB - Magnetic resonance imaging (MRI) provides high-resolution morphological images useful in diagnostic radiology to differentiate between normal and abnormal/pathological states in tissues. More recently, emerging developments in MRI have added a functional/physiological dimension to anatomical images. Electron paramagnetic resonance (EPR), a magnetic resonance technique similar to nuclear magnetic resonance, detects paramagnetic species such as free radicals. Like MRI, EPR can be implemented as an imaging technique for small animals and potentially human applications. Because of the low abundance of naturally occurring paramagnetic species, exogenous paramagnetic species are needed for in vivo EPR imaging (EPRI). The image data from EPRI contain both spatial distribution of paramagnetic species and spectral information. Hence, spatially encoded functional information such as tissue oxygen status and redox status can be extracted and coregistered with the spatial distribution of the spin probe, to the anatomy, or both by suitable means. Ultimately, the images obtained from EPRI may be used to overlay the functional information (containing spatial tissue physiology information) onto detailed anatomical maps. With its ability to enable whole animal imaging in mice, EPRI will be a useful imaging technique that complements other techniques such as MRI and positron emission tomography in obtaining valuable functional/physiological images. PMID- 11406721 TI - Use of positron emission tomography in animal research. AB - Among the several imaging technologies applied to in vivo studies of research animals, positron emission tomography (PET) is a nuclear imaging technique that permits the spatial and temporal distribution of compounds labeled with a positron-emitting radionuclide to be determined noninvasively. It can be viewed as an in vivo analog of classic autoradiographic methods. Many different positron labeled compounds have been synthesized as tracers that target a range of specific markers or pathways. These tracers permit the measurement of quantities of biological interest ranging from glucose metabolism to gene expression. PET has been extensively used in imaging studies of larger research animals such as dogs and nonhuman primates. Now, using newly developed high-resolution dedicated animal PET scanners, these types of studies can be performed in small laboratory animals such as mice and rats. The entire whole-body biodistribution kinetics can be determined in a single imaging study in a single animal. This technique should enable statistically significant biodistribution data to be obtained from a handful of animals, compared with the tens or hundreds of animals that might be required for a similar study by autoradiography. PET also enables repeat studies in a single subject, facilitating longitudinal study designs and permitting each animal to serve as its own control in experiments designed to evaluate the effects of a particular interventional strategy. This paper provides a basic overview of the methodology of PET imaging, a discussion of the advantages and drawbacks of PET as a tool in animal research, a description of the latest generation of dedicated animal PET scanners, and a review of a few of the many applications of PET in animal research to date. PMID- 11406722 TI - Ultrasound imaging: principles and applications in rodent research. AB - Ultrasound imaging utilizes the interaction of sound waves with living tissue to produce an image of the tissue or, in Doppler-based modes, determine the velocity of a moving tissue, primarily blood. These dynamic, real time images can be analyzed to obtain quantitative structural and functional information from the target organ. This versatile, noninvasive diagnostic tool is widely used and accepted in human and veterinary medicine. Until recently its application as a research tool was limited primarily to larger, nonrodent species. Due to advances in ultrasound imaging technology, commercially available ultrasound systems now have the spatial and temporal resolution to obtain accurate images of rat and mouse hearts, kidneys, and other target tissues including tumor masses. As a result, ultrasound imaging is being used more frequently as a research tool to image rats and mice, and particularly to evaluate cardiac structure and function. The developing technology of ultrasound biomicroscopy has even greater spatial resolution and has been used to evaluate developing mouse embryos and guide site specific injections into mouse embryos. Additional ultrasound imaging technologies, including contrast-enhanced imaging and intravascular ultrasound transducers adapted for transesophageal use, have been utilized in rats and mice. This paper provides an overview of basic ultrasound principles, equipment, and research applications. The use of noninvasive ultrasound imaging in research represents both a significant refinement as a potential replacement for more invasive techniques and a significant advancement in research techniques to study rats and mice. PMID- 11406723 TI - Challenges in small animal noninvasive imaging. AB - The current status and challenges of small animal non-invasive imaging is briefly reviewed. The advantages of non-invasive studies on living animals versus post mortem studies are evaluated. An argument is advanced that even in post-mortem situations, non-invasive imaging may play an important role in efficiently characterizing small animal phenotypes as well as pathology. Issues of data interpretation under anesthetized conditions in live animal studies are also reviewed. The five imaging technologies covered include CT, PET, ultrasound, MRI and optical imaging. The structural and physiological information content of these different modalities is reviewed along with the ability of these techniques to scale down for use in small mammals such as mice and rats. In general, it was found that most of these technologies scale favorably to the study of small mammals, generally providing more physiological information than when used on the larger human scale. This suggests that these types of small mammal imaging capabilities will play a very significant role in the full utilization of these important animal models in biomedical research. PMID- 11406724 TI - Platelet aggregation inhibition with glycoprotein IIb--IIIa inhibitors. AB - Inhibitors of the platelet receptor glycoprotein (GP) IIb--IIIa are a novel and potent class of antithrombotic drugs for the management of patients with non-ST segment elevation acute coronary syndromes (ACS) and those undergoing percutaneous coronary intervention (PCI). Pharmacodynamic studies with three currently approved agents in this class (abciximab [ReoPro, Centocor, Inc., Malvern, Pennsylvania, and Eli Lilly & Company, Indianapolis, Indiana]; eptifibatide [INTEGRILIN, COR Therapeutics, Inc., South San Francisco, California, and Key Pharmaceuticals, Inc., Kenilworth, New Jersey]; and tirofiban HCI [Aggrastat, Merck & Co., Inc., Whitehouse Station, New Jersey]) all sought to identify dosing regimens that would establish and maintain >80 % inhibition of ex vivo platelet aggregation throughout the duration of intravenous infusion. Direct comparison of these prior studies is difficult, however, because the assays used different anticoagulants (sodium citrate [abciximab, tirofiban HCI] or D phenylalanyl-L-prolyl-L-arginine chloromethyl ketone [PPACK] [eptifibatide]) and different concentrations of the platelet agonist adenosine diphosphate (ADP) (5 micromol [tirofiban HCI] or 20 micromol [abciximab, eptifibatide]). More recent work has attempted to overcome these limitations by using similar assay conditions for all GP IIb--IIIa inhibitors. These studies have indicated that the concentrations of all three agents required to provide the targeted effect for platelet inhibition are considerably higher in the presence of an anticoagulant that does not chelate calcium ions (e.g., heparin or PPACK) than in the presence of calcium-chelating anticoagulant (i.e., sodium citrate). Of the three currently approved GP IIb--IIIa inhibitors, eptifibatide is the only agent whose approved dosing is based on an ex vivo platelet aggregation assay that uses such an anticoagulant. Additionally, Kereiakes et al. have recently reported that the high levels of platelet inhibition (>80 %), using PPACK as an anticoagulant and ADP (20 micromol) as an agonist, are more consistently achieved with the approved dosing regimen of eptifibatide. The antiplatelet effect of abciximab showed more interpatient variability, whereas the median inhibition of ex vivo platelet aggregation with the approved dosing regimen for tirofiban HCl was <80 % at almost all time points during drug infusion. PMID- 11406725 TI - Factor V R506Q mutation-Leiden: an independent risk factor for venous thrombosis but not coronary artery disease. AB - BACKGROUND: A specific point G-A transition at nucleotide position 1691 in the factor V (FV) gene, FV-Leiden, was associated with increased risk of venous thromboembolism (VTE). Insofar as the association of FV-Leiden with coronary artery disease (CAD) remains poorly defined, the aim of this study was to determine the prevalence of FV-Leiden in a sample of 68 VTE patients, 69 CAD patients, and 192 randomly selected healthy subjects. METHODS: Total genomic DNA was extracted from the peripheral blood of study subjects and was used for PCR analysis. The presence (or absence) of FV-Leiden was assessed by PCR using primers flanking the mutant site (nt 1691), followed by hybridization with wild type ('G') and mutant ('A') biotinylated DNA probes; detection was by DNA enzyme immunoassay (DEIA). RESULTS: While the prevalence of FV-Leiden in CAD patients was not statistically different from that of healthy subjects (14.5 % vs. 15.1 %; P=0.890, odds ratio 0.95; 95 % confidence interval 0.43--2.06), a significant increase in FV-Leiden prevalence was seen in VTE patients (70.6 % in VTE patients; P<0.001, odds ratio 13.4, 95 % confidence interval 6.9--25.8). Of the 48 VTE patients who tested positive for FV-Leiden, 42 were heterozygotes (G/A), while 6 were homozygotes (A/A) (allele frequency 0.397). All 10 CAD patients positive for FV-Leiden were heterozygote carriers (allele frequency 0.072). While gender was not a factor in FV-Leiden expression, higher prevalence in FV-Leiden was seen in younger (< or =45 years) VTE patients (38/51 vs. 10/17). CONCLUSION: FV-Leiden is a major inherited risk factor for VTE, with a peak incidence in younger patients, but does not appear to play any role in CAD pathogenesis in the population studied. PMID- 11406726 TI - Aspirin in patients with coronary artery disease: is it simply irresistible? PMID- 11406727 TI - Thrombolysis for restoration of patency to haemodialysis central venous catheters: a systematic review. AB - Urokinase, previously used to restore patency to thrombosed haemodialysis catheters, is now unavailable in North America. We performed systematic reviews of four questions related to the safety and efficacy of alternative agents for catheter thrombolysis, searching Medline and the Cochrane Controlled Clinical Trials Register. In dialysis patients, large case series have documented that urokinase is safe and effective (>70 % efficacy for catheter instillation, and >80 % for systemic lysis). Experience with streptokinase is limited and allergic complications develop with repeated use. Studies of catheter instillation with 1- 2 mg of tPA per lumen reported short-term success in 83--98 % of uses. One non peer-reviewed study described 44--59 % success using systemic tissue plasminogen activator (tPA), 2.5 mg through each of 2 lumens, over 1 h. Meta-analysis of randomized comparisons of urokinase and tPA as full-dose thrombolytic agents suggested that 1 mg tPA was likely equivalent in thrombolytic potency to 36,000 units urokinase. In nondialysis populations, four case series suggested that catheter instillation with 0.5--2 mg tPA was effective and safe in reestablishing patency, and a randomized controlled trial found 2--4 mg tPA more effective than 5,000--10,000 units urokinase. No complications have been reported in any patient treated with systemic or local tPA for catheter thrombolysis. In studies of fistula thrombolysis with 5--50 mg tPA major complications occurred in one episode in 130 patients treated. This review suggests that 1--2 mg/lumen tPA is a suitable dose for catheter instillation and likely to be more effective than 5000 units/lumen urokinase. Systemic lysis with 5--10 mg tPA is likely to be safe and effective in suitably selected patients. Further studies are needed. PMID- 11406728 TI - Thrombosis in Behcet's disease: report of a case followed by a systematic review using the methodology of evidence-based medicine. AB - BACKGROUND: Behcet's disease (BD) is a rare systemic inflammatory disease of unknown aetiology with a variety of organ manifestations. METHODS: A case of Behcet's disease complicated by deep vein thrombosis (DVT) and inferior vena cava thrombosis referred to our institution is reported. In addition, a structured literature search using the methodology of evidence-based medicine for the localization of venous lesions, treatment modalities, and outcome of patients with Behcet's disease and concurrent thrombosis was performed. A modified validity score was assigned by established methods. RESULTS: In total, 214 citations were identified using our search strategy. Among these citations, 10 papers including a total of 32 patients (25 male, 7 female) met the inclusion criteria and were incorporated into this overview together with our patient. All studies consisted of serial case reports without control subjects. Neither localization of venous lesions, nor treatment modalities were homogeneous not allowing general recommendations. CONCLUSIONS: No data are available from controlled studies regarding treatment modalities of patients presenting with Behcet's disease complicated by concurrent thrombosis. This lack of evidence implicates the need for large scale and co-ordinated registries including data on the acute treatment as well as the prevention of future thrombotic events in this clinical setting. The diagnostic criteria of the "International Study Group for Behcet's Disease" may well serve as a basis for this approach. PMID- 11406729 TI - The hemostatic effects of warfarin titration in post CABG patients in comparison to placebo treatment. AB - BACKGROUND: Since coronary artery bypass graft patients remain at risk of coronary artery and bypass graft occlusion after successful surgery, adjunct treatment regimens are under investigation. In a study of the patients of the multicenter Post Coronary Artery Bypass Graft (Post CABG) Trial, 1 mg warfarin was found to have no important effect on coagulation parameters. STUDY DESIGN: The effects of 1, 2 and 3 mg warfarin were evaluated at six-week intervals in 20 Post CABG Trial patients receiving titrated dose increases in comparison to 20 patients of similar age, gender and time from CABG treated with placebo. RESULTS: International normalized ratio (INR) values increased with warfarin dose increments for 1, 2, and 3 mg, respectively (0.95+/-0.16, 1.08+/-0.19, and 1.34+/ 0.39) and in comparison to placebo treated patients (dosextreatment p<0.001). Factor VII coagulant activity decreased with warfarin titration (1 mg, 119.0+/ 18.3 %; 2 mg, 100.6+/-32.8 %; 3 mg, 95.0+/-27.8 %) and in comparison to placebo (dosextreatment p=0.008). Levels of prothrombin fragment F1.2, tissue plasminogen activator, fibrinogen and von Willebrand factor were unchanged with warfarin dose increments and in comparison to placebo. CONCLUSIONS: At doses up to 3 mg, warfarin acts on the INR through a reduction of factor VII with no effect on the fibrinolytic system, fibrinogen or von Willebrand factor. At these doses F1.2 did not document reduced coagulation activity. The observations of this study were consistent with the decision in the Post CABG Trial to increase the warfarin dose above 1 mg to achieve a distinct effect of warfarin that was less than full anticoagulation. PMID- 11406730 TI - Clinical application of procedural platelet monitoring during percutaneous coronary intervention among patients at increased bleeding risk. AB - The goal of platelet function testing in the catheterization laboratory is to provide information about the platelet contributions to the risk of thrombotic or hemorrhagic events and optimization of anti-platelet therapy for percutaneous interventions. We present several illustrative cases in which platelet monitoring with the Rapid Platelet Function Assay (RPFA, Accumetrics) was used to guide dosing of a glycoprotein (GP) IIb/IIIa inhibitor for coronary and peripheral intervention among patients at increased bleeding risk. PMID- 11406731 TI - Usefulness of combining necrosis and platelet markers in triaging patients presenting with chest pain to the emergency department. AB - BACKGROUND: Myocardial injury and platelet activation play important roles in the pathogenesis of unstable coronary syndromes. We sought to determine whether the combined measurement of platelet and necrosis markers would improve risk stratification, and yield higher diagnostic utility in patients presenting to the emergency department with chest pain. METHODS AND RESULTS: Platelet and soluble P selectin together with myoglobin, creatine kinase, CK-MB fraction, and troponin I were measured from the autologous samples in 122 consecutive patients. Statistical analysis revealed strong Spearman correlation coefficients (0.141- 0.412; p<0.001) between platelet expression of P-selectin and plasma levels of necrosis markers. Platelet P-selectin and necrosis markers were independent predictors (c-index>0.7) for acute myocardial infarction, while plasma P-selectin exhibited random distribution. Elevated soluble P-selectin and myoglobin were the most valuable in identifying patients with congestive heart failure. None of the markers were useful for triaging chest pain patients with unstable angina. Analysis of incremental gains (Chi-squares) reveals that with respect to platelet P-selectin, myoglobin adds 50 % to AMI diagnostic value, and creatine kinase yields an additional 20 % in triaging these patients. The diagnostic value of soluble P-selectin is substantially (72 %) increased by myoglobin measurements, and enhanced even further (44 %) by adding cardiac troponin I for identifying heart failure patients among the chest pain population. CONCLUSION: Simultaneous determination of platelet and necrosis markers improve the early diagnosis of acute myocardial infarction and congestive heart failure among patients with chest pain presenting into the Emergency Department. Well controlled clinical trials are needed to prove the advantage of combining platelet and necrosis data over presently used techniques in emergency medicine. PMID- 11406732 TI - Aspirin and ACE-inhibitors: for wedding or funeral? PMID- 11406733 TI - The role of low-molecular-weight heparins in the management of unstable angina and non-ST-segment elevation myocardial infarction. PMID- 11406735 TI - Clinical pharmacology: niche or panorama? And does it matter? PMID- 11406734 TI - Anaphylactoid reactions to vitamin K. AB - Anaphylactoid reactions in patients receiving intravenously administered vitamin K have been reported in the literature. To summarize the known data on anaphylactoid reactions from administration of vitamin K, we reviewed all published and unpublished reports of this adverse reaction. Published reports were obtained through medline (1966--1999) and EMBASE (1971--1999) searches of the English language literature and review of references from identified case reports. Unpublished reports were obtained using the Spontaneous Reporting System Adverse Reaction database of the United States Food and Drug Administration (FDA) between August 1968 and September 1997. All adverse drug reactions to vitamin K were categorized by route of drug administration, dose and standard adverse reaction code. In the FDA reports, we defined anaphylactoid reactions as any adverse drug reaction coded as either anaphylaxis, allergic reaction, apnea, dyspnea, death, heart arrest, hypotension, shock or vasodilatation. Additionally, all fatal and life-threatening FDA reported reactions were reviewed to determine if they could represent an anaphylactoid reaction missed by the above definition. The literature review uncovered a total of 23 cases (3 fatal) of anaphylactoid reactions from intravenous vitamin K. The FDA database contained a total of 2236 adverse drug reactions reported in 1019 patients receiving vitamin K by all routes of administration. Of the 192 patients with reactions reported for intravenous vitamin K, 132 patients (69 %) had a reaction defined as anaphylactoid, with 24 fatalities (18 %) attributed to the vitamin K reaction. There were 21 patients with anaphylactoid reactions and 4 fatalities reported with doses of intravenous vitamin K of less than 5 mgs. For the 217 patients with reactions reported due to vitamin K via a non-intravenous route of administration, 38 patients had reactions meeting the definition of anaphylactoid (18 %), with 1 fatality (3 %) attributed to the drug. The absolute risk of an anaphylactoid reaction to intravenous vitamin K cannot be determined by this study, but the relatively small number of documented cases despite widespread use of this drug suggest that the reaction is rare. Anaphylactic reactions and case fatality reports were found even when intravenous vitamin K was given at low doses by slow dilute infusion. The pathogenesis of this reaction is unknown and may be multifactorial with etiologies including vasodilation induced by the solubilizing vehicle or immune-mediated processes. We conclude that use of intravenous vitamin K should be limited to patients with serious hemorrhage due to a coagulopathy that is secondary to a relative or absolute deficiency of vitamin K. PMID- 11406736 TI - Integration of pharmacokinetic and pharmacodynamic studies in the discovery, development, and review of protein therapeutic agents: a conference report. PMID- 11406737 TI - Effects of rifampin on the pharmacokinetics and pharmacodynamics of glyburide and glipizide. AB - OBJECTIVE: To study the effects of rifampin (INN, rifampicin) on the pharmacokinetics and pharmacodynamics of glyburide (INN, glibenclamide) and glipizide, 2 sulfonylurea antidiabetic drugs. METHODS: Two separate, randomized, 2-phase, crossover studies with an identical design were conducted. In each study, 10 healthy volunteers received 600 mg rifampin or placebo once daily for 5 days. On day 6, a single dose of 1.75 mg glyburide (study I) or 2.5 mg glipizide (study II) was administered orally. Plasma glyburide and glipizide and blood glucose concentrations were measured for 12 hours. RESULTS: In study I, rifampin decreased the area under the plasma concentration--time curve [AUC(0-infinity)] of glyburide by 39% (P <.001) and the peak plasma concentration by 22% (P =.01). The elimination half-life of glyburide was shortened from 2.0 to 1.7 hours (P <.05) by rifampin. The blood glucose decremental AUC(0-7) (net area below baseline) and the maximum decrease in the blood glucose concentration were decreased by 44% (P =.05) and 36% (P <.001), respectively, by rifampin. In study II, rifampin decreased the AUC(0-infinity) of glipizide by 22% (P <.05) and shortened its half-life from 3.0 to 1.9 hours (P =.01). No statistically significant differences in the blood glucose concentrations were found between the phases; however, 4 subjects had moderate hypoglycemia during the placebo phase but only 1 subject had moderate hypoglycemia during the rifampin phase. CONCLUSIONS: Rifampin moderately decreased the plasma concentrations and effects of glyburide but had only a slight effect on glipizide. The mechanism underlying the interaction between rifampin and glyburide is probably induction of either CYP2C9 or P-glycoprotein or both. Induction of CYP2C9 would explain the increased systemic elimination of glipizide. It is probable that the blood glucose- lowering effect of glyburide is reduced during concomitant treatment with rifampin. In some patients, the effects of glipizide may also be reduced by rifampin. PMID- 11406738 TI - Population pharmacokinetic analysis of pegylated interferon alfa-2b and interferon alfa-2b in patients with chronic hepatitis C. AB - BACKGROUND: This study quantified pharmacokinetic changes in pegylated and nonpegylated interferon alfa-2b during 48 weeks of treatment and the influences of covariates on the basis of sparsely sampled serum concentrations and activity values. Possible relationships between pharmacokinetic and pharmacodynamic variables were investigated. METHODS: Patients with chronic hepatitis C were enrolled in a clinical trial that compared the efficacy of pegylated interferon alfa-2b with interferon alfa-2b. Single blood samples were obtained from each patient at weeks 4, 12, 24, 36, and 48. Three pharmacostatistical models were developed for 2 immunoassays and 1 bioassay. RESULTS: Apparent clearance values of pegylated interferon alfa-2b and interferon alfa-2b at the end of treatment declined 33.7% and 80.0%, respectively, from their week 4 values. Bioactivity increased 41% to 58% at week 48 for different treatment groups. Changes were greatest in the first weeks of administration and diminished during the subsequent months. Body weight had a modest positive effect on clearance values and activity. Within each dose level, no significant associations were observed between pharmacokinetic variables and any pharmacodynamic variables (hepatitis C virus--RNA responses or changes in neutrophils and platelets). CONCLUSIONS: This analysis confirms earlier observations of progressive pharmacokinetic changes in the patients with hepatitis C during 48 weeks of treatment. The absence of a relationship between toxicity or efficacy variables and interferon concentration or activity (within a dose level) suggests that clinical management of patients (eg, for efficacy or to manage toxicity) should be based on clinically derived dosing guidelines rather than on serum concentration or activity criteria. PMID- 11406739 TI - Removal of digoxin by column for specific adsorption of beta(2)-microglobulin: a potential use for digoxin intoxication. AB - BACKGROUND: A beta(2)-microglobulin adsorption column used for the treatment of dialysis-related amyloidosis removes serum beta(2)-microglobulin by recognition of lipophilic residue in the protein. No data are available for the adsorption of the highly lipophilic drug digoxin. METHODS: In vivo clearance of digoxin with the beta(2)-microglobulin column was measured by a single use of the column in 8 patients receiving hemodialysis with a therapeutic level of digoxin. In vitro adsorption was evaluated by use of incubation with adsorbent of the column and digoxin or ranitidine, a hydrophilic drug. Clearance with the beta(2) microglobulin column was further compared with that obtained by use of activated charcoal in the dogs intoxicated with digoxin. RESULTS: Digoxin concentration was reduced from 1.11 +/- 0.25 ng/mL to 0.57 +/- 0.15 ng/mL at 240 minutes after initiation of hemoperfusion with the column in the patients. Digoxin clearance with the beta(2)-microglobulin column was about 145 +/- 20 mL/min, with a blood flow rate of 160 to 220 mL/min (80% of plasma flow rate). Eighty-five percent of digoxin was adsorbed in vitro, and the capacity of the beta(2)-microglobulin column was not saturated until a toxic level was reached (50 ng/mL). This value was higher than that obtained with use of charcoal. In dogs with digoxin intoxication, digoxin clearance was 38.9 +/- 1.5 mL/min, with a blood flow rate of 50 mL/min (95% of plasma flow rate), which was almost twice as that achieved with charcoal. The degree of thrombocytopenia and leukopenia was small with use of the beta(2)-microglobulin column. CONCLUSION: These data suggested that the beta(2)-microglobulin column selectively adsorbs digoxin. This column is a promising tool for the treatment of digoxin intoxication, especially in patients undergoing hemodialysis. PMID- 11406740 TI - The sugar absorption test in the evaluation of the gastrointestinal intolerance to bisphosphonates: studies with oral pamidronate. AB - RATIONALE AND AIMS Some bisphosphonates induce gastrointestinal side effects, but the localization in the gastrointestinal tract and the underlying mechanism are unknown. The feasibility of the sugar absorption test was investigated to assess the gastrointestinal effects of oral enteric-coated pamidronate. The sugar absorption test measures the urinary excretion of lactulose, mannitol, and sucrose after oral intake. Increases in the lactulose/mannitol ratio and sucrose excretion indicate increased small intestinal permeability and gastroduodenal disease, respectively. SUBJECTS AND METHODS: Twelve volunteers (5 women and 7 men) participated in a randomized, double-blind, 4-way crossover study. The sugar absorption test was performed 2 hours after the final drug intake following a 3 day course of enteric-coated pamidronate (300 mg daily), placebo, or acetylsalicylic acid (3 g daily). The lactulose/mannitol ratio and sucrose excretion were measured in urine collected for 5 hours after ingestion of the solution. The fourth treatment consisted of intravenous administration of pamidronate. Treatment comparison was with paired t tests after log transformation. RESULTS: The lactulose/mannitol ratio after pamidronate and acetylsalicylic acid administration was 54% and 118% higher than that after placebo (95% confidence intervals [CI], +8%, +119%, and +69%, +182%). The lactulose/mannitol ratio after pamidronate administration was 29% lower (95% CI, 54%, +3%) than that after acetylsalicylic acid. Compared with placebo the sucrose excretion was 290% higher after acetylsalicylic acid (95% CI, +46%, +518%) but only 8% higher after pamidronate (95% CI, -41%, +97%). The absorption of pamidronate was below 1%, and there was no relationship with the increased lactulose/mannitol ratio. CONCLUSION: Oral enteric-coated pamidronate increases intestinal but not gastroduodenal permeability. There was no relationship between intestinal permeability and absorption of pamidronate. It appears that the sugar absorption test is an appropriate, noninvasive method for evaluation of gastrointestinal effects of bisphosphonates in humans. PMID- 11406741 TI - Bupropion SR as an aid to smoking cessation in smokers treated previously with bupropion: a randomized placebo-controlled study. AB - BACKGROUND: Many persons who attempt to quit smoking have made previous unsuccessful attempts to quit with pharmacologic aids. An understanding of the impact of these previous attempts to quit is vital for selecting medications that may be more successful in a future attempt to quit. In particular, the effect of repeated use of bupropion SR (Zyban; INN, amfebutamone) on abstinence rates has not been studied previously. METHODS: This was a multicenter, randomized, double blind, placebo-controlled study in 450 smokers who had previously used bupropion in a smoking cessation attempt. The study consisted of a screening phase, a 12 week treatment phase, and a follow-up at month 6. Participants made regular clinic visits throughout the treatment phase during which they received brief counseling sessions to encourage abstinence from smoking. The primary end point was continuous abstinence from smoking from weeks 4 through 7. Secondary efficacy end points were examined throughout the treatment phase and at follow-up after 6 months. RESULTS: In participants receiving bupropion SR, 27% (61 of 226) remained abstinent throughout the period from weeks 4 through 7 compared with 5% (11 of 224) of participants receiving placebo (P <.001). Significantly (P <.001) more participants who received bupropion SR during the treatment phase remained continuously abstinent from the start of week 4 through month 6 (27 of 226; 12%) compared with participants who received placebo (5 of 224; 2%). Eleven participants receiving placebo (5%) and 19 participants receiving bupropion SR (8%) stopped taking the study medication because of an adverse event. CONCLUSIONS: Bupropion SR is an effective medication for retreatment of smokers who have used bupropion SR previously. PMID- 11406742 TI - Comparison of patient questionnaires, medical records, and plasma assays in assessing exposure to benzodiazepines in elderly subjects. AB - OBJECTIVE: Exposure in pharmacoepidemiologic studies can rely on various sources such as medical records, patient questionnaires, or plasma samples, which do not always concur. This study endeavored to compare sources of information on current exposure to benzodiazepines in elderly subjects. METHODS: In a study in a hospital admissions department, 1136 elderly subjects included in a case-control study each completed a structured questionnaire. In addition, an inspection of the medical records of each subject was performed, as well as screening of a plasma sample (high-pressure liquid chromatography--diode array detector) for current exposure to benzodiazepines. RESULTS: Benzodiazepines were found in the plasma of 33% of 1013 patients, in the records of 31% of patients, and in the questionnaires of 36% of 797 respondents. With use of the plasma results as a standard, questionnaires had 11% false positives and 28% false negatives; medical records had 14% false positives and 23% false negatives. The kappa for concordance between questionnaires and records was 0.63. Most of the errors were related to the unexpected presence in plasma of clorazepate, commonly used as a hypnotic agent. CONCLUSIONS: Patient recall and medical records are not reliable measures of current exposure to benzodiazepines in elderly persons, although this unreliability may be more marked with certain drugs used as hypnotic agents. PMID- 11406743 TI - Drug interactions as a cause of overanticoagulation on phenprocoumon or acenocoumarol predominantly concern antibacterial drugs. AB - BACKGROUND: The risk of hemorrhage when coumarin anticoagulants are used sharply increases when the international normalized ratio (INR) is > or = 6.0. Such overanticoagulation may be caused by drug interactions. We performed a case control study among previously stable outpatients of an anticoagulation clinic using phenprocoumon or acenocoumarol to identify changes in the use of potentially interacting drugs related to overanticoagulation. METHODS: Three hundred case patients with INR values > or = 6.0 were compared with 302 randomly selected matched control subjects with INR values within the target zone. Information on changes in the use of 87 potentially interacting drugs in the 4 weeks before the index day was collected by interviewing patients and by reviewing the anticoagulant medical record. RESULTS: Forty-five potentially interacting drugs were not used in the 4-week study period, and only 15 drugs were used by at least 10 patients. For a number of drugs, too few patients had a relevant change in use to judge their association with overanticoagulation. A course of a combination product of sulfamethoxazole and trimethoprim strongly increased the risk of overanticoagulation (adjusted odds ratio, 24.2; 95% confidence interval [CI], 2.8 to 209.1; population attributable risk percentage [PAR%], 5.7%), especially in patients receiving acenocoumarol. Penicillins were associated with a risk of overanticoagulation of 2.4 (95% CI, 1.00 to 5.5); the corresponding PAR% was 3.4%. The effect was confined to amoxicillin (INN, amoxicilline) plus clavulanic acid. CONCLUSION: Drug interactions as a cause of overanticoagulation predominantly concerned antibacterial drugs. If possible, the use of sulfamethoxazole-trimethoprim and amoxicillin plus clavulanic acid should be avoided in patients receiving coumarins. If there is no therapeutic alternative available, increased monitoring of INR values is warranted to prevent overanticoagulation and potential bleeding complications. PMID- 11406744 TI - Incidence of serious adverse drug reactions in general practice: a prospective study. AB - BACKGROUND: Many studies have been conducted to estimate the incidence and economic impact of adverse drug reactions. Most of these studies used historical data or were based on single hospital units. Little is known, however, about the frequency of serious adverse drug reactions in general practice. OBJECTIVE: To estimate the incidence of serious adverse drug reactions in the community. METHODS: A prospective study during 5 consecutive working days between March 1 and April 30, 1998, was conducted among a random representative sample of 254 general practitioners in Aquitaine, France. The main outcome measure was the number of serious adverse drug reactions (ie, resulting in death, life threatening condition, hospitalization, incapacity, or sequel) observed by each general practitioner during the study period and validated by an expert panel. RESULTS: Thirteen validated serious adverse drug reactions, 2 of which were fatal (1 subarachnoidal hemorrhage with oral anticoagulant and 1 aplastic anemia with antineoplastics), were observed, resulting in an incidence density of 10.2 (95% confidence interval [CI], 5.4 to 17.5) per 1000 days of practice. Eleven case subjects (84.6%) were hospitalized. This represents an average of 2.6 cases per general practitioner per year, and 123,000 adverse drug reaction cases (95% CI, 65,400 to 210,000) for the 60,000 general practitioners in France. Antineoplastics and anticoagulants were the drugs most frequently involved, and blood dyscrasia and bleeding were the most frequent adverse drug reactions. CONCLUSION: This study, which is one of the few available that has prospectively measured the incidence of serious adverse drug reactions in general practice settings, confirms that serious adverse drug reactions are a major public health concern. PMID- 11406745 TI - Limited sampling approach. PMID- 11406750 TI - Management of fever in neutropenic patients. AB - Substantial progress has been made in the management of febrile episodes in neutropenic patients, largely by the prompt administration of potent, broad spectrum antimicrobial agents. During the past several decades, the spectrum of organisms has changed from a predominance of gram-negative pathogens to a predominance of gram-positive pathogens. In recent years, some hospitals have experienced an increase of infections caused by multi-drug-resistant pathogens. Hence, it is no longer possible to rely on standardized regimens, but antimicrobial therapy must be selected based on the predominant pathogens and antimicrobial susceptibility patterns at each institution. It is customary to initiate antifungal therapy empirically in those patients whose fever persists despite broad-spectrum antibacterial therapy. Alternatives now exist to amphotericin B, including lipid formulations of this drug, and fluconazole. It is critically important that each patient be carefully re-assessed before starting antifungal therapy, because there are many other potential causes for persistent fever, including resistant bacteria and viruses. Novel approaches to therapy include outpatient antibiotics, and use of growth factors as adjunctive therapy. There also has been a renewed interest in white blood cell transfusions. Although the prognosis for infection in neutropenic patients has improved greatly, new infectious problems have emerged that limit our successful management of these complications. PMID- 11406751 TI - Discovery of the causative organism of Weil's disease: historical view. AB - On January 20, 1915, Inada and Ido announced the discovery of the causative agent of Weil's disease. Subsequently, on February 13, 1915, they published the first paper on the discovery of the causative organism (a new species of Spirochaeta) of Weil's disease. Besides discovering the causative organism of the disease, Inada and colleagues clarified the pure culture in medium, and determined the source and route of the infection, its pathology and morbid anatomy; the distribution of the organism in various organs and tissues; the excretion of the spirochete, and its division, filterability, and morphological characteristics; and the clinical picture, laboratory findings, diagnosis, prophylaxis, and treatment of the disease. These studies were conducted by Inada, Ido, Kaneko, Hoki, and Ito, in the years 1914 to 1915. In the early investigation of leptospirosis, Inada and colleagues played a prominent part. We would like to remember these remarkably complete and definitive original achievements on leptospirosis made by Inada and colleagues. PMID- 11406752 TI - Comparison of antimycobacterial activity of grepafloxacin against Mycobacterium avium with that of levofloxacin: accumulation of grepafloxacin in human macrophages. AB - The bactericidal activity of two new quinolones, grepafloxacin and levofloxacin, against five strains of Mycobacterium avium was investigated in vitro. The minimum inhibitory concentrations (MICs) of these two quinolones, determined by the broth microdilution method, were comparable for all strains tested. In contrast, grepafloxacin suppressed the intracellular growth of all the strains in monocyte-derived macrophages more strongly than levofloxacin, when the cells infected with these strains were incubated for 7 days in the presence of various concentrations of the two new quinolones. To find the reason for the strengthened intracellular killing activity of grepafloxacin, we determined the ratio of the concentration of the new quinolones in the cells to that in the medium (C/M concentration ratio). The C/M concentration ratio of grepafloxacin was increased to 34.7 by 7 days, whereas that of levofloxacin at 7 days was only 12.3. These data suggested that a higher level of intraphagocytic accumulation of grepafloxacin endows it with greater mycobactericidal activity. PMID- 11406753 TI - Characterization of MRSA transmission in an emergency medical center by sequence analysis of the 3'-end region of the coagulase gene. AB - The distribution of methicillin-resistant Staphylococcus aureus (MRSA) isolates at the St. Marianna University affiliated emergency medical center (EMC) was studied by sequence analysis of the 3'-end region of the coagulase gene. We collected a total of 42 MRSA isolates, consisting of 20 strains from the hospital environment, 13 strains from the nostrils or fingers of medical staff, and 9 strains from inpatients in the EMC. We compared our results with those from 27 stock strains of known coagulase serotype and 2 strains reported in the literature. All 69 strains tested have four to six tandem repeats in the 3'-end region of the coagulase gene. Among the 42 MRSA isolates collected, the base sequence of the 3'-end region of the coagulase gene was identical in 28 of them (67%). The number of isolates originating from the hospital environment, medical staff, and patients, respectively, that were identical to this representative strain were 18 (90%), 6 (46%), and 4 (44%). Phylogenetic analysis using the DNA sequences of the tandem repeat region demonstrated that almost all strains from the patients formed a concordant cluster with the representative strain from the hospital ward. We also assessed the value of sequence analysis of the 3'-end region of the coagulase gene as an epidemiological marker. Our results indicate that sequence analysis of the 3'-end region of the coagulase gene of MRSA may be a potent epidemiologic typing system. PMID- 11406754 TI - Increase of R5 HIV-1 infection and CCR5 expression in T cells treated with high concentrations of CXCR4 antagonists and SDF-1. AB - The chemokine receptors CXCR4 and CCR5 are considered to be potential targets for the inhibition of HIV-1 replication. We found that the synthetic peptides T134 and T140 (see text for full names) inhibited X4 HIV-1 infection with selectivity and low toxicity because they acted as CXCR4 antagonists. However, high concentrations of T134, T140, and ALX40-4C (see text for full name) increased the expression of CCR5 and R5 HIV-1 infection, as did stromal cell-derived factor 1 (SDF-1). In contrast to CXCR4 antagonists and SDF-1, viral monocyte inflammatory protein (vMIP) II inhibited not only anti-CXCR4 monoclonal antibody (MAb) but also inhibited anti-CCR5 MAb binding to human peripheral blood mononuclear cells, and inhibited both X4 and R5 HIV-1 strains. T134, T140, ALX40-4C, and SDF-1 increased viral transcription in the treated cells. In addition, ALX40-4C and SDF 1 also increased nuclear transcription factor (NF)-kappaB. However, the mechanisms of action of T134 and T140 are different from those of clinically used anti-HIV drugs. Thus, synergistic activities were observed in the concomitant treatment with T134 and reverse transcriptase inhibitors or protease inhibitors. Our findings, presented here, are noteworthy in regard to the potential clinical use of these agents as drugs for the treatment of AIDS. PMID- 11406755 TI - Comparison of chest X-ray findings and other parameters in acute exacerbation of chronic bronchitis in Japan and the West. AB - One of the issues concerning harmonization in the development of pharmaceutical products, especially antimicrobials, is discrepancy in the indications to be studied clinically. In particular, it has been very much questioned whether the underlying disease in Western patients diagnosed with acute exacerbation of chronic bronchitis (AECB) is identical with chronic bronchitis in Japan. We assessed chest X-ray films from 105 AECB patients enrolled in a clinical study of SB265805 (a fluoroquinolone antibacterial agent under development) conducted in Europe, and then compared their clinical signs/symptoms and laboratory data with Japanese historical data. Five of the 105 patients did not meet the criteria of AECB; i.e., 2 of them were diagnosed with pneumonia, 1 with bronchiectasis, 1 with pneumoconiosis, and 1 with bronchiectasis plus pulmonary emphysema. In the remaining 100 patients, chest X-ray findings and laboratory test results were consistent with the concept of chronic bronchitis, although 23 of them had other cardiac or pulmonary diseases as well. There were significant imbalances in distribution between Western patients and Japanese historical data in terms of age, cough, WBC counts, and C-reactive protein (CRP) levels. Compared with Japanese historical data, Western patients were younger and had a more severe cough, although increases in WBC and CRP were less remarkable. For other variables, i.e., sex, fever, and volume of sputum, no significant difference was detected in distribution. Overall, there was no significant difference between the two groups in regard to disease severity, as assessed by fever, WBC, and CRP. PMID- 11406756 TI - Nosocomial fungemia due to amphotericin B-resistant Candida spp. in three pediatric patients after previous neurosurgery for brain tumors. AB - Amphotericin B (AmB) resistance in Candida spp. is very rare. Three cases of fungemia, due to amphotericin B-resistant Candida spp. in pediatric patients after previous neurosurgery for brain tumors, are reported. The Candida strains - one C. guillermondii, one C. lusitaniae, and one C. parapsilosis - showed minimum inhibitory concentrations (MICs) to AmB of 2-4 microg/ml. Two of the three patients had been pretreated with AmB for 5-11 days. All three patients were successfully treated with intravenous fluconazole (6-10 mg/kg per day) for 16-28 days, and all survived. Despite AmB resistance in Candida spp. being very rare, C. lusitaniae, C. guillermondii, and C. parapsilosis isolates in documented infections should be tested for AmB resistance, mainly in patients not responding to therapy with AmB. PMID- 11406757 TI - Emergence of cephem- and aztreonam-high-resistant Neisseria gonorrhoeae that does not produce beta-lactamase. AB - Regarding Neisseria gonorrhoeae, the National Committee for Clinical Laboratory Standards (NCCLS) has not defined the breakpoint minimum inhibitory concentration (MIC) for expanded spectrum cephems such as cefpodoxime and ceftizoxime, because of the absence of resistant strains to these antibiotics. To date, in gonococcal urethritis, after treatment with third generation cephems and aztreonam, clinical failures caused by resistant N. gonorrhoeae strains have not been reported. However, we experienced two clinical failures in patients with gonococcal urethritis treated with cefdinir and aztreonam. N. gonorrhoeae isolates from these two patients showed high-level MICs to these agents. The MIC of cefdinir was 1 microg/ml for both strains and that of aztreonam was 8 microg/ml for both strains, while the MICs of other beta-lactams were also higher than the NCCLS value, except for ceftriaxone, for which the MIC was 0.125 microg/ml for both strains. Moreover, the MICs of fluoroquinolones, tetracyclines, and erythromycin against these two isolates were higher than the NCCLS susceptibility value. These isolates were susceptible to spectinomycin. In N. gonorrhoeae, the emergence of these beta-lactam-resistant isolates is of serious concern. However, a more serious problem is that these isolates were already resistant to non-beta-lactam antimicrobials. In Japan, ceftriaxone has not been permitted for clinical use against gonococcal infections. Therefore, in Japan, patients with gonococcal urethritis caused by these resistant N. gonorrhoeae strains should be treated with cefodizime or spectinomycin. PMID- 11406758 TI - Postantibiotic suppression effect of macrolides on the expression of flagellin in Pseudomonas aeruginosa and Proteus mirabilis. AB - The phenomenon of postantibiotic effect (PAE) encompasses not only the effects of bacterial growth inhibition but also the suppression of virulence factors. We tentatively designated the latter effect the postantibiotic suppression effect (PASE). The flagella of Gram-negative bacteria are involved in the development of biofilms. We measured the PASE of erythromycin (ERY) and azithromycin (AZM) on the expression of flagellin in Pseudomonas aeruginosa and Proteus mirabilis. Flagellin preparations were subjected to sodium dodecylsulfate (SDS) polyacrylamide gel electrophoresis (PAGE) analysis and the flagellin band was identified by N-terminal amino acid sequence analysis. We thus evaluated the flagellin by the intensity of the band. The mean durations of the PAE of ERY and AZM on bacterial growth were 0.9 and 2.0 h for P. mirabilis, and 0.6 and 2.7 h for P. aeruginosa, respectively. The PASE of these drugs on flagellin expression was also observed. The apparent PASEs of ERY and AZM on flagellin were up to 5 h for P. mirabilis and up to 6 h for P. aeruginosa after a single 0.5 x minimum inhibitory concentration (MIC) treatment for 5 h. Our results suggest that certain combinations of antibiotics may have prolonged suppressive effects on the expression of virulence factors in certain Gram-negative bacteria. PMID- 11406759 TI - Incidence of deep fascial space infection after surgical removal of the mandibular third molars. AB - Nine hundred and ninety-three patients who underwent surgical removal of the mandibular third molars with oral antibiotic prophylaxis were examined to determine the incidence of postoperative deep fascial space infection and its background factors. Postoperative deep fascial space infection was observed in 8 of the patients (0.8%; 4 males and 4 females), and submandibular spaces were involved in all infected patients. Only 1 of these 8 patients was an immune compromised host. Patients aged 30 years or more had a significantly higher incidence of deep fascial space infection than those aged under 30. Five patients had partial bony impactions and 3 had complete bony impactions. However, the incidence of infection according to the molar positions was not significantly different between partial bony impaction and complete bony impaction. The 8 patients had not had pericoronitis preoperatively. The clinical courses of all were favorable after antibiotics were administered intravenously. In conclusion, the incidence of deep fascial space infection after removal of the mandibular third molars was low, at 0.8%. However, it may be desirable to remove the molars, if applicable, at a younger age because of the higher incidence of infection in patients aged over 30. The results of this study also offer information that will be useful as a basis for obtaining informed consent from patients whose mandibular third molars are to be removed. PMID- 11406760 TI - Localized adenovirus gene delivery using antiviral IgG complexation. AB - Gene therapy with viral vectors has progressed to clinical trials. However, the localization of viral vector delivery to diseased target sites remains a challenge. We tested the hypothesis that an adenoviral vector could be successfully delivered by complexation with a specific antibody that is bound to a biodegradable matrix designed for achieving localized gene transduction. We report the first successful delivery system based upon antibody immobilization of virions in a type I collagen-avidin gel using a polyclonal biotinylated IgG specific for the adenovirus hexon. In vitro stability studies demonstrated retention of viral vector activity with antibody-complexed adenovirus collagen gel preparations, in comparison to loss of vector activity from collagen gels prepared with nonspecific biotinylated IgG. Cell culture investigations using this antibody-controlled release system for adenoviral vector transduction of rat aortic smooth muscle cells (A10) demonstrated a significantly more localized reporter expression (beta-galactosidase) compared with non-antibody-complexed controls. Herpes simplex thymidine kinase (HSVtk) adenoviral vectors were immobilized on avidin-collagen gels via this antibody-complexation approach, and ganciclovir was added to rat smooth muscle cells (A10) in culture with the gels. With complexed HSVtk adenovirus, only cells either in contact with the virus containing gel or within 50 microm were killed. By comparison, at the same adenovirus and ganciclovir dose, non-antibody-complexed HSVtk adenoviral delivery with ganciclovir resulted in the death of virtually all cells. Myocardial gene transfer studies in pigs demonstrated significantly more efficient right ventricular adenoviral GFP expression with anti-hexon antibody-complexed matrix injections, compared with direct vector injections. Thus, our results show that matrix formulations based on antibody-complexation delivery of adenovirus resulted in site-specific localization of transgene expression that enhances the efficiency of therapeutic vector strategies and provides a potent means for localization, to avoid distal side-effects. This approach has therapeutic potential as an implantable preparation that through the means of antibody complexation, can localize and optimize viral vector gene therapy. PMID- 11406761 TI - Wild-type p53 gene transfer inhibits neointima formation in human saphenous vein by modulation of smooth muscle cell migration and induction of apoptosis. AB - Patency of autologous human saphenous vein coronary artery bypass grafts (CABG) is compromised by intimal thickening and superimposed atherosclerosis, caused by migration of vascular smooth muscle cells (SMC) to the intima where they proliferate. Here, using adenoviral transfer, we have targeted SMCs using wild type p53 (wt p53) overexpression. Initial in vitro analyses demonstrated that wt p53 overexpression had no effect on SMC proliferation but promoted apoptosis, which was inhibited by co-expression of bcl2 or crmA. Wt p53 inhibited SMC invasion through reconstituted matrices, a phenotype not affected by bcl2 or crmA. Overexpression of wt p53 in human saphenous vein before organ culture significantly induced apoptosis (P < 0.01, Student's t test) without affecting proliferation rates either in the media or in the intima. SMC migration was, however, significantly reduced by wt p53 (P < 0.01, Student's t test). Intimal thickening and the number of neointimal cells were reduced by 89% and 73%, respectively, after 14 days (P < 0.01 and P < 0.001, respectively, Student's t test). This study demonstrates that overexpression of wt p53 promotes apoptosis and inhibits migration of SMC leading to reduced intimal thickening. This maybe a useful approach for increasing patency rates in CABG procedures in the clinic. PMID- 11406762 TI - Interaction between DNA-cationic liposome complexes and erythrocytes is an important factor in systemic gene transfer via the intravenous route in mice: the role of the neutral helper lipid. AB - Recent studies have indicated that there are many barriers to successful systemic gene delivery via cationic lipid vectors using the intravenous route. The purpose of this study was to investigate the effect of binding and interaction between erythrocytes, a major constituent of blood cells, and the complexes, in relation to the role of the helper lipid, on the in vivo gene delivery to the lung following intravenous injection. We used three types of cationic lipid vectors, DNA-DOTMA/Chol liposome complexes, DNA-DOTMA liposome complexes, and DNA DOTMA/DOPE liposome complexes. Although the three types of vectors bind to murine blood cells in vivo and in vitro, DOTMA/Chol and DOTMA complexes with a higher in vivo transfection activity do not induce fusion between erythrocytes, whereas DOTMA/DOPE complexes, a less efficient vector in vivo, induce fusion between the erythrocytes after a short incubation period. Pre-incubation of DOTMA/DOPE complexes with erythrocytes significantly reduced the transfection efficiency while DOTMA/Chol- and DOTMA complexes were more resistant to such treatment. The differences in the physicochemical and structural properties of these complexes could explain the differences in interaction with erythrocytes and subsequent gene expression. Lipids in DOTMA/Chol and DOTMA complexes have a stable lamellar structure. However, lipids in DOTMA/DOPE complexes have a highly curved structure with high fluidity. These results indicate that the interaction with erythrocytes depends on the properties of the cationic lipid vectors and this is an important factor for intravenous gene delivery using cationic lipid vectors. PMID- 11406763 TI - Efficient retrovirus-mediated transduction of primitive human peripheral blood progenitor cells in stroma-free suspension culture. AB - Retroviral transduction of hematopoietic cells has resulted in unsatisfactory gene marking in clinical studies. Since cytokine-stimulated stem cells have engrafted poorly in animal models, we investigated phenotypic changes during culture of peripheral blood progenitor cells (PBPC). Human CD34(+) HLA-DR(low) cells, immunomagnetically separated from PBPC collections, were found to extrude rhodamine-123, which is characteristic for primitive hematopoietic cells. Cells were grown in suspension cultures supplemented with cytokines. While interleukin 3-containing factor combinations promoted cell proliferation they caused loss of rhodamine-123 extrusion and reduced the frequencies of cobblestone area-forming cells (CAFC). Several other cytokines failed to stimulate cell divisions, which are required for retroviral transduction. A combination including Flt-3 ligand (FL), interleukin-6 and stem cell factor (SCF) preserved an immature phenotype for 5 to 6 days and stimulated cell divisions, which was improved upon addition of leukemia inhibitory factor and interleukin-11. Furthermore, the CAFC frequency among cells treated with these cytokines was increased as compared with widely used cocktails containing interleukin-3, interleukin-6 and SCF. Rhodamine-123 appeared to be a particularly sensitive indicator for differentiation of PBPC. For analysis of gene transfer, amphotropic retroviruses conferring an MDR1 cDNA were added repeatedly for 6 days to cytokine-treated PBPC stroma-free cultures. Proviral cDNA was detected by polymerase chain reaction in 68% of cobblestone areas derived from CD34(+)HLA-DR(low) cells that had been exposed to Flt-3 ligand, interleukin-6 and SCF. In summary, conditions were identified that facilitate efficient transduction of early PBPC with amphotropic retroviruses while preserving a primitive phenotype for extended periods. PMID- 11406764 TI - A novel human suspension culture packaging cell line for production of high-titre retroviral vectors. AB - Retroviruses are currently the most widely used vectors in clinical trials for gene therapy. These vectors are, however, limited by low titres partly due to the restrictive nature of monolayer cell culture. We have developed a stable suspension producer cell line derived from human lymphoblastoid cells (WIL-2) by electroporating these cells with the necessary trans components required for production of defective retrovirus particles which encode a nuclear localising beta-galactosidase gene. We show that this anchorage-independent cell line generates viruses at a titre of 7 x 10(5) iu/ml on NIH3T3 indicator cells which remains constant after at least 2 months in culture. The producer cells can be cultured at a density of 6 x 10(6) cells/ml with consistent virus titre production. WIL-2 can also be grown as single cells by rotation culture while maintaining virus production. By treating the cells with the transcriptional activator sodium butyrate titres above 1 x 10(6) i.u./ml are achieved. Concentrating viral supernatants by ultrafiltration can further increase virus titre to 5 x 10(8) i.u./ml. Even at these high titres no replication-competent virus was detected. Virus titre fell only slightly when cells were placed in serum-free media before harvest. The generation of this novel cell line provides proof-of-principle that large-scale production of retroviral vectors in serum free growth conditions can be safely generated for use in gene therapy. PMID- 11406765 TI - Recombinant adenovirus expressing adeno-associated virus cap and rep proteins supports production of high-titer recombinant adeno-associated virus. AB - It has been difficult to produce a chimeric vector containing both Ad and AAV rep and cap, and to grow such chimeric vectors in 293 cells. By recombination in vitro in a bacterial host, we were able to produce recombinant plasmid AdAAV (pAdAAVrep-cap), which could be used to generate recombinant AdAAV (rAdAAVrep cap) after transfection into 293 cells. A recombinant adenovirus, rAdAAVGFP, in which the green fluorescent protein (GFP) gene is flanked by the AAV terminal repeats cloned into the E1-deleted site of Ad was also generated. Co-infection of rAdAAVrep-cap together with rAdAAVGFP into 293 cells resulted in production of high titers of rAAV expressing GFP. It was noted that the titer of rAdAAVrep-cap was lower than the titer of control AdCMVLacZ. The lower titer of rAdAAvrep-cap was associated with expression of Rep protein. Non-homologous recombination occurs after high passage and results in deletions within the AAV rep genes. These results indicate that (1) rAdAAVrep-cap can be produced; (2) rAdAAVrep-cap + rAdAAVGFP is a convenient and efficient way to transfect 293 cells to grow high titer rAAV; and (3) frozen stock is required to avoid propagation of rep-deleted pAdAAVrep-cap. PMID- 11406766 TI - Triggered intracellular activation of disulfide crosslinked polyelectrolyte gene delivery complexes with extended systemic circulation in vivo. AB - We have developed polyelectrolyte gene delivery vectors that display good extracellular stability and are activated intracellularly to permit transgene expression. The strategy comprises covalent crosslinking of primary amines in poly-L-lysine/DNA complexes with a crosslinking agent that can later be cleaved by reduction. Crosslinked complexes maintained the same size and surface charge but showed increased stability against polyelectrolyte exchange with poly-L aspartic acid. Surface modification with polyethyleneglycol improved solubility and masked their positive surface charge. Crosslinked complexes showed 10-fold increased plasma circulation following intravenous administration to Balb/c mice. In the absence of chloroquine, the levels of transgene expression in B16F10 murine melanoma cells were similar for crosslinked and non-crosslinked complexes, however, chloroquine selectively potentiated transgene expression by the non crosslinked complexes. Cellular uptake of the complexes was the same, irrespective of crosslinking. Following microinjection into the cytoplasm of Xenopus oocytes, or the cytoplasm or nucleus of Rat-1 fibroblasts, crosslinked complexes mediated the same transgene expression as non-crosslinked complexes, indicating crosslinked complexes are rapidly reduced and activated intracellularly. We therefore hypothesize that the lower in vitro transfection activity of crosslinked complexes in the presence of chloroquine is due to reduced transfer from endosome to cytoplasm, mainly due to increased stability against destabilization by chloroquine. The extended systemic circulation together with triggered intracellular activation makes these complexes a promising system for targeted gene delivery in vivo. PMID- 11406767 TI - A synthetic leucine zipper-based dimerization system for combining multiple promoter specificities. AB - One of the biggest challenges facing gene therapy is the development of vectors that direct the activity of therapeutic genes specifically to the sites of disease. To achieve this goal, the restriction of transgene transcription via synthetic promoters that are endowed with multiple specificities represents a particularly promising strategy. Towards this end, we have developed a generally applicable strategy (DCTF system) where a synthetic promoter is driven by an artificial heterodimeric transcription factor whose DNA-binding and transactivating subunits are expressed from two promoters with different selectivity. A crucial determinant of the DCTF system is the heterodimerization interface that should provide for a high affinity interaction without interference by endogenous proteins. Here, we describe such a dimerization system based on engineered Fos and Jun leucine zippers. We show the usefulness of this system for the combination of cell type-specific and cell cycle-regulated transcription and demonstrate its functionality in an in vivo setting. PMID- 11406768 TI - A simplified system for constructing recombinant adenoviral vectors containing heterologous peptides in the HI loop of their fiber knob. AB - The use of recombinant adenovirus (Ad) vectors containing genetically modified capsid proteins is an attractive strategy for achieving targeted gene transfer. The HI loop of the fiber knob is a promising candidate location for the incorporation of foreign ligands for achieving this goal. However, the method of constructing an Ad vector containing a foreign ligand in the HI loop of the fiber knob has proved difficult. In this study, we developed a simple system to construct fiber-modified vectors. To do this, a vector plasmid containing a complete E1/E3-deleted Ad type 5 genome and a unique Csp45I and/or ClaI site between positions 32679 and 32680 of the Ad genome (residues threonine-546 and proline-547 of the fiber protein) was constructed. Oligonucleotides corresponding to the Arg-Gly-Asp (RGD) or Asn-Gly-Arg (NGR)-containing peptide motif (as a model) and containing a Csp45I and/or ClaI recognition site, were ligated into the Csp45I and/or ClaI-digested plasmid. The foreign transgene expression cassette was inserted into the E1 deletion site of the vector plasmid and the fiber-mutant Ad vector was produced by transfection of the PacI-digested plasmid into 293 cells. The virus containing the RGD or NGR peptide on the fiber knob was able to infect human glioma cells, which do not express coxsackievirus and adenovirus receptor (CAR), one of the Ad virus receptors, about 100-1000 times more efficient than the virus containing wild-type fiber. This suggested that the mutant virus mediated CAR-independent cell entry pathway. The simplicity of this method allows not only for easy construction of fiber-mutant Ad vectors, but also for screening of the peptides that target the vector to the desired cells and tissues. PMID- 11406771 TI - Developing a UK health equity network. AB - The UK Health Equity Network (HEN) was established in 1999 to encourage and facilitate multidisciplinary and multi-stakeholder collaboration in health inequalities research. This paper briefly details the history, objectives and location of HEN. PMID- 11406772 TI - Domestic violence: a comparative survey of levels of detection, knowledge, and attitudes in healthcare workers. AB - The objective of this study was to compare the knowledge, attitudes, responses and levels of detection of domestic violence among a variety of healthcare workers in different specialities.Self-administered questionnaires were sent to community and hospital based healthcare workers in Oxfordshire working in primary care, obstetrics and gynaecology, mental health and accident and emergency. These comprised all principal general practitioners and general practitioner registrars, 50% of practice/district nurses and health visitors in each practice, and all healthcare workers in obstetrics and gynaecology, community mental health teams and accident and emergency in one trust. The amount of domestic violence detected in different healthcare settings was far less than indicated by anonymous surveys and crime figures. Knowledge about many of the issues surrounding domestic violence was inconsistent and there were fundamental deficiencies. The attitudes of healthcare workers to domestic violence were generally sympathetic and supportive. Women, nurses and community mental health workers reported significantly better knowledge and more positive attitudes than other respondents. Gender, role and speciality were independently associated with more positive attitudes and the latter two were independently associated with good knowledge. The response that healthcare workers make when they uncover domestic violence is confused and often inappropriate. In conclusion, most healthcare workers accept that domestic violence is a healthcare issue but lack fundamental knowledge about the issues surrounding domestic violence itself and appropriate agencies that can offer help. They also lack skills in identifying and discussing this issue with patients/clients. A large, unfulfilled training need has been identified. PMID- 11406773 TI - The importance of injury as a cause of death in sub-Saharan Africa: results of a community-based study in Tanzania. AB - This paper describes rates and causes of injury deaths among community members in three districts of the United Republic of Tanzania. A population-based study was carried out in two rural districts and one urban area in Tanzania. Deaths occurring in the study areas were monitored prospectively during a period of six years. Censuses were conducted annually in the rural areas and biannually in the urban area to determine the denominator populations. Cause-specific death rates and Years of Life Lost (YLL) due to injury were calculated for the three study areas. During a 6 year period (1992-1998), 5047 deaths were recorded in Dar es Salaam, 9339 in Hai District and 11 155 in Morogoro Rural District. Among all ages, deaths due to injuries accounted for 5% of all deaths in Dar es Salaam, 8% in Hai and 5% in Morogoro. The age-standardised injury death rates among men were approximately three times higher than among women in all study areas. Transport accidents were the commonest cause of mortality in all injury-related deaths in the three project areas, except for females in Hai District, where it ranked second after intentional self-harm. We conclude that injury deaths impose a considerable burden in Tanzania. Strategies should be strengthened in the prevention and control of avoidable premature deaths due to injuries. PMID- 11406774 TI - How useful are sociodemographic characteristics in identifying children at risk of unintentional injury? AB - The social class gradient in childhood injury mortality is steep and increasing, so there is emphasis on targeting injury prevention on the basis of socioeconomic deprivation, to reduce inequalities in health. This paper examines the relationship between medically attended unintentional injury, sociodemographic characteristics and previous injury. This was a cohort study using the control group from a cluster randomised controlled trial of injury prevention in primary care. The cohort comprised children aged 3-12, months registered with participating practices, whose parents completed the baseline questionnaire (n = 771). 94% were followed for 25 months. Medically attended unintentional injury was ascertained from the primary and secondary care records. Logistic regression analysis examined the relationship between sociodemographic factors, previous injury and the occurrence of future medically attended injuries. Poisson regression examined the relationship between sociodemographic factors, previous injury and the number of future medically attended injuries. The response rate to the questionnaire was 75%. Residence in a deprived ward, lack of access to a car and male sex were associated with at least one medically attended injury. Residence in a deprived ward and young maternal age were associated with hospital admission. Residence in a deprived ward, male sex and non-ownership of a car were independently associated with number of unintentional injuries. Specificity exceeded sensitivity for all factors for medically attended injury and hospital admission. The positive predictive value was low for all factors, especially for hospital admissions. In conclusion, residence in a deprived ward was independently associated with any medically attended injury, with hospital admission and with number of injuries received. However, more than half of those children residing in a deprived ward did not have a medically attended injury and more than 90% did not have a hospital admission. 60% of children who had a medically attended injury and 40% who had a hospital admission do not live in a deprived ward. A combination of a population approach and targeted interventions will achieve the greatest health gain, and is unlikely to widen inequalities in health. PMID- 11406775 TI - Evaluating maternity services: how representative are data collected using a pregnancy diary? AB - As a part of the evaluation of Nottingham's response to the Changing Childbirth initiative, a cohort of pregnant women were asked to complete diaries, recording their perceptions of events and their feelings over the entire pregnancy period. Before the study began, we were concerned that the diary completion rate could be low and that those completing would be unrepresentative of the population. In this paper, we report the nature of responses to the diary and show that these problems were far less serious than was initially expected. A high degree of coverage was achieved, implying that large numbers of women were sufficiently motivated to maintain detailed records over many months. The research suggests that the diary is a viable method of data capture in such circumstances. PMID- 11406776 TI - Assessment of HIV transmission risks in clinical situations in health care students from Germany, Lithuania and Namibia. AB - Due to the increasing global HIV prevalence, comprehensive knowledge about clinical HIV transmission risks in health care professionals is essential. Mainly medical and nursing students are at risk, because they are working close to infected patients. By using an anonymous questionnaire, the study analysed the assessments of German medical/dental students (n = 182), Lithuanian medical students (n = 176) and Namibian student nurses (n = 135) on the risk of getting HIV infected in different clinical situations. It became obvious that the Namibian student nurses overestimated the risk of HIV transmission in several situations (eg changing dirty linen, physical examination). In comparison, the Lithuanian students showed the most realistic assessments, while the German students also tended to overestimate the risks of HIV transmission. The results indicate that assessments on the risk of HIV transmission in clinical situations are influenced by the national prevalence and daily contacts with HIV patients. Education of health care students should consider this and focus more on epidemiological aspects and infection control procedures, to avoid endangering students and patients. PMID- 11406777 TI - The efficacy of whole cell pertussis immunisation: collected data on a vaccine produced in France. AB - The place of whole cell pertussis vaccines in paediatric immunisation schedules is under re-evaluation by public health authorities in many countries, with the expectation that the newly licensed acellular Bordetella pertussis vaccines will induce fewer adverse events while providing equivalent efficacy. In France, for instance, the CSHPF (Conseil superieur d'hygiene public de France) recently modified its long-standing recommendation that French children only receive whole cell pertussis vaccine. Consequently, an acellular pertussis vaccine may be used for the first booster, at 16-18 months of age, and should be used for the reinforcing dose at 11-13 y of age. French children, nonetheless, continue to receive whole cell pertussis vaccine for the primary series immunisations at 2, 3, and 4 months, as the only whole cell pertussis vaccine available in France (licensed by Aventis Pasteur) has a long-established record of safety and protective efficacy. A review of its unpublished and published clinical results, obtained from studies throughout the world, demonstrates an efficacy of from 84 100% in six different retrospective analyses or outbreak investigations and a protective efficacy of 92% by clinical trial. PMID- 11406778 TI - Acute flaccid paralysis surveillance in Southern Italy. AB - The purpose of this study was to report the results of the first two years' surveillance programme of acute flaccid paralysis (AFP) in Southern Italy (Calabria). All paediatric, neurology and infectious diseases wards from 23 hospitals were selected. Stool and serum samples to determine the presence of poliovirus were collected. Throat swabs were taken within 10 days of onset of illness. During the period March 1997-April 1999 eight cases of AFP were reported and four of them, three females and one male, occurred in children younger than 15 y of age, although none was confirmed as poliomyelitis. The rate of non-polio AFP in 1997 and 1998 was, respectively, 0.24 and 0.73 per 100 000 persons under 15 y of age. Our results demonstrated that an active surveillance has permitted us to immediately detect AFP cases and to exclude those due to wild polio virus and to vaccine-associated cases and indicated that our area seems to be 'polio free'. PMID- 11406779 TI - Relationships between dietary intake and cognitive function level in Korean elderly people. AB - We examined the relationship between dietary intake and cognitive performance in Korean elderly people. Data for dietary intake, anthropometric measurements and cognitive function tests were collected and the relationships of the variables were analyzed. A random sample of 210 men and 239 women in Korea, aged 60 and over, was selected. Subjects were free-living elderly people who had not experienced major cognitive function impairment. Main outcome measures, 24 h dietary recall method, food behaviour variables, anthropometrics indices, health variables, and Kwon's Mini-Mental State Examination for Koreans (MMSE-K) for cognitive function test. The prevalence rate of poor cognitive function (MMSE-K score < or = 19) of Korean elderly was 22.3%: women with poor cognitive function had a higher rate (31.0%) than that in men (12.3%). Cognitive ability was related negatively with age and positively with school education level. Female subjects of poor cognitive function had significantly lower intakes of total amount of foods, cereals, vegetables, fruits, milk, spices, and also, energy, protein, fat, carbohydrate, Ca, P, Fe, vitamin A, thiamin, riboflavin, and niacin than those of the normal cognitive score (> or = 24) group (P < 0.05). Male subjects of poor cognitive function had significantly lower intakes of fruits, fiber, and vitamin C than the normal subjects (P < 0.05). The MMSE-K score of female subjects showed a significant positive correlation with total amount of foods, cereals, beans, fruits, milk, oil, spices, and energy, protein, fat, carbohydrate, Ca, Fe, P, riboflavin and niacin intakes. The consumption of adequate nutrients, by taking sufficient amounts and variety of foods, may be important in maintaining adequate cognitive function in elderly Koreans. PMID- 11406780 TI - Perceptual strategies to improve skin cancer discriminations in naive observers. AB - The aim of these experiments was to assess the effect of an educational brochure on the ability of naive observers to discriminate skin cancers from benign lesions, and to investigate possible new strategies to assist observers in performing this task. A two-alternative forced choice paradigm was used to investigate the ability of observers to correctly identify different types of benign and malignant lesions before and after exposure to an educational brochure. The method of pair comparisons was used to assess the ability of observers to discriminate between benign and malignant lesions under different instruction conditions. Subjects were undergraduate students from Sydney University. An educational brochure did not facilitate the ability to correctly identify malignant lesions, and appeared to result in deterioration of performance in the identification of benign lesions. Similarly, observers were unable to discriminate between benign and malignant lesions on the basis of how dangerous they looked. However, judgements of lesions in terms of perceived distinctiveness resulted in relatively accurate discriminations between benign and malignant lesions. These data suggest current skin cancer detection strategies may be ineffective in improving the ability to visually identify benign and malignant lesions. Discriminating between lesions in terms of how distinctive they appear may form the basis of a new and effective strategy for the detection of skin cancer. PMID- 11406781 TI - Effects of lifestyle factors on ultrasonographically determined bone health in Japanese women. AB - The objective of this study was to determine by ultrasonographic measurements, an inexpensive and radiation-free technique, the association between bone health and lifestyle factors among a large population of Japanese women. Two hundred and fifty-six pre-menopausal women and 585 post-menopausal women who underwent a voluntary medical check-up for osteoporosis in 1996-1997 were analyzed. There were significant positive correlations between the bone density (designated as the stiffness value) vs the weight, the height and the body mass index of the subjects only in the post-menopausal group. Negative correlations were also found between the bone density vs the age and the years since menopause. Our data using ultrasonographic technique agree well with previous studies using other devices. In both groups, subjects with current or past exercise habits had higher stiffness values. Dietary habits had no effects on the stiffness value. Smoking habits had a trend towards negative effects and alcohol consumption seemed to have a trend towards positive effects on the stiffness value in post-menopausal women, but these effects did not reach statistical significance. Positive effects of current exercise on bone density were maintained after adjustment for past exercise habits. These results support the effectiveness of exercise begun in adulthood. Having a good exercise habit is one of the most effective ways of maintaining good bone health. PMID- 11406782 TI - Comparative anthropometrics of 3 to 24-month-old children breastfed in both high and low socio-economic strata in Lagos, Nigeria. AB - This is a cross-sectional study of infants seen in the Nitel Health Centres in Lagos metropolis and Mushin Primary Health Care Centres in the Mushin Local Government Area of Lagos State. A total of 297 infants aged 3-24 months were studied. Anthropometric and breast feeding patterns of the children were studied. The mean weights of the mothers were 67.2 kg for Nitel mothers and 62.11 kg for the mothers in Mushin Primary Health Care Centres. The mean heights for the mothers were 71.47 cm and 66.1 cm, respectively. About 98.8% of the mothers in the Primary Health Care Centres of Mushin breastfed. About 7.4% of the total population studied had WAZ scores of less than -2s.d. while 12.8% had HAZ scores of less than -2s.d. and 7.43% had WHZ scores of less than -2s.d. Malnutrition was considered to be very high. There was a positive correlation between the anthropometric measurement and education. PMID- 11406787 TI - Does the perception of downloading speed influence the evaluation of web-based lectures? AB - Medical schools put little emphasis upon education on public health, even though public health has played an important role in this century. One way to harness its benefit in order to improve global health in the 21st century is to globally share lectures on public health through the Internet. We have developed the Supercourse comprising of web-based learning modules on epidemiology in a standardized format with the size of each web page less than 10 kilobytes. A cross-sectional observational study was conducted to investigate the association of the perception of the access speed to Web-based lectures by teachers with their perception of lecture quality. There were 223 teachers who rated the lectures: 72% were from North America or Western Europe, 40% had taught epidemiology, and 14% reported that the speed of access was slow. Odds ratio of above-average rating among those who reported that the speed of access was fast relative to those who reported that the speed of access was slow was 4.25 (2.03 8.91; P = 0.001). The odds ratios were similar and significant after taking into account several other factors, including the variation of rating across lectures, region, and experiences in teaching epidemiology. The results indicate that the perception of the quality of Web-based lectures is related to the speed of access to a web page. The speed of access may be as important, if not more important, as the content itself. This suggests that, to share educational materials on the Web globally for teachers, one must consider not only the content, but also how people at local sites gain access to the Internet. PMID- 11406788 TI - A crisis looms on the horizon. PMID- 11406789 TI - A prospective 5-year study of fixed partial prostheses supported by implants with machined and TiO2-blasted surface. AB - PURPOSE: The aim of the present study was to evaluate whether there was a difference between machined and TiO(2)-blasted implants regarding survival rate and marginal bone loss during a 5-year observation period. MATERIALS AND METHODS: A total of 133 implants (Astra Tech Dental Implants; Astra Tech AB, Molndal, Sweden) were placed in 50 patients at 6 centers in 4 Scandinavian countries. Forty-eight implants were installed in the maxilla and 85 implants in the mandible. A randomization and a stratification were done, so that each fixed partial prosthesis was supported by at least 1 machined and 1 TiO(2)-blasted implant. The implant-supported fixed partial prostheses (ISFPP) were fabricated within 2 months after postoperative healing. A total of 52 ISFPP (17 maxillary, 35 mandibular) were inserted. The patients were clinically examined once a year for 5 years. At the annual follow-up, biological as well as technical complications were recorded. RESULTS: Of the 133 implants placed, 3 were reported as failed after 5 years of follow-up, resulting in an overall cumulative survival rate of 97.6%. The cumulative implant survival rates were 100% for the TiO(2) blasted implants and 95.1% for the machined implants. No significant difference in survival was, however, found between the machined and TiO(2)-blasted implants after 5 years. The mean marginal bone loss in the maxilla was 0.21 +/- 0.83 mm (SD) for the machined implants and 0.51 +/- 1.11 mm (SD) for the TiO(2)-blasted implants during the 5-year observation period. In the mandible, the mean marginal loss was 0.22 +/- 1.13 mm for the machined implants and 0.52 +/- 1.07 mm for the TiO(2)-blasted implants from baseline to the 5-year examination. No significant difference in marginal bone loss between the 2 surface groups was found during the 5-year observation period. CONCLUSIONS: The present study shows good 5-year results with small ISFPP in the mandible, as well as in the maxilla. No significant differences were found in failure rate and marginal bone loss around implants with a machined rather than a TiO(2)-blasted surface. J Prosthodont 2001;10:2-7. PMID- 11406790 TI - Wear of enamel opposing low-fusing and conventional ceramic restorative materials. AB - PURPOSE: This study evaluated the wear area of human enamel opposing 2 conventional and 2 low-fusing dental porcelains, as well as abrasive wear, attrition, surface hardness, and fracture toughness for the 4 porcelain substrates. MATERIALS AND METHODS: Two low-fusing and 2 conventional metal ceramic porcelains were used to form 15-mm-diameter disks (n = 10), which were fired according to manufacturer's recommendations. Enamel cusps (n = 40) were formed from extracted third molars. All ceramic and enamel specimens were finished to a 1000-grit silicon carbide surface. The Oregon Health Sciences University (Portland, OR) oral wear simulator was used to deliver a 20-N load from the cusps to the ceramic substrates through a food-like slurry. The sliding action of the cusps over an 8-mm linear path produced abrasive wear. A static 70 N load was applied at the end of the path to create attrition. This sequence was repeated at 1.0 Hz for 50,000 cycles. Ceramic wear was measured with a profilometer (+/-2 micrometers), and enamel wear was evaluated using optical scanning methods. After wear testing, the hardness and fracture toughness of the ceramic surfaces were determined, and scanning electron photomicrographs were made using representative ceramic and enamel specimens from each group. Enamel wear areas, abrasion and attrition depths, hardness, and fracture toughness values were subjected to analysis of variance and Tukey-Kramer post hoc tests to determine significant differences. RESULTS: Enamel wear was not significantly different for low-fusing and conventional porcelains (p =.29). The wear of conventional and low-fusing ceramic substrates was also not significantly different (p =.79). However, depth of porcelain wear caused by attrition was in general significantly greater than abrasive wear (p =.0004). Although no significant differences were found in the measured porcelain hardness values (p =.08), 1 conventional porcelain exhibited fracture toughness significantly greater than 1 low-fusing porcelain (p <.01). CONCLUSIONS: No differences in wear patterns were noted among low-fusing compared with conventional metal-ceramic porcelains, but static loading resulted in significantly greater attrition compared with the observed sliding abrasive wear. J Prosthodont 2001;10:8-15. PMID- 11406791 TI - Stresses within porcelain veneers and the composite lute using different preparation designs. AB - PURPOSE: The purpose of this study was to investigate compressive and tensile stresses in porcelain and composite at the labial marginal region of porcelain veneer restorations using chamfer, shoulder, or knife-edge labial margin designs with labial window or incisal overlap incisal preparation designs. METHODS: Porcelain veneer models were constructed and loaded with (1) a 200-N, 45 degrees palatal load to simulate functional loading, and (2) a horizontal labial loading of 200 N to mimic trauma. Maximum tensile and compressive stresses were recorded within the labial marginal region of both porcelain and composite lute. RESULTS: Under the 45 degrees palatal load, stresses within the palatal marginal porcelain were chiefly compressive, and stresses for the knife-edge designs as much as 42% less than for shoulder designs. Incisal overlap preparations were generally associated with less compressive stress within both porcelain and composite than the window preparation. When a labial load was applied, tensile stresses were as much as 25 times greater for the chamfer and shoulder designs compared with the knife-edge design. Labial loading also resulted in an increase in tensile stresses within the composite lute, and stresses were again lowest within the knife-edge margin design. CONCLUSIONS: Under the limitations of this study, using the incisal overlap preparation, porcelain veneers with knife-edge labial margins could better sustain occlusal loading without fracture. J Prosthodont 2001;10:16 21. PMID- 11406792 TI - Effect of abutment tooth reinforcement techniques on the fracture resistance of removable partial denture master casts. AB - PURPOSE: The purpose of this study was to compare the mean loads required to fracture an isolated stone abutment tooth when various reinforcement techniques were used: (A) no reinforcement, (B) surface hardening, (C) impression trimming, (D) dowel placement, and (E) impression trimming plus dowel placement. MATERIALS AND METHODS: Die stone premolar specimens for each group (n = 4) were loaded perpendicular to the tooth long axis in 100-g increments 25 hours after pouring the impression. Mean load to failure was determined for each group. Data were evaluated with a one-way analysis of variance (alpha = 0.05) to determine differences among groups. Paired comparisons of factor level means were made with Tukey's q-test. RESULTS: The mean fracture load for Group E (7.6 kg) was significantly greater than Groups A (2.0 kg), B (2.1 kg), and C (3.4 kg). No significant difference was found between Groups D (6.0 kg) and E. CONCLUSIONS: Dowel reinforcement with impression trimming at the cervical portion of the abutment tooth resulted in significantly greater fracture resistance than the control, surface model hardener, or impression trimming alone. J Prosthodont 2001;10:22-25. PMID- 11406793 TI - In vitro comparison of the fracture resistance and failure mode of fiber, ceramic, and conventional post systems at various stages of restoration. AB - PURPOSE: This in vitro study evaluated 6 post systems over 4 simulated clinical stages of tooth restoration to (1) determine quantitatively the fracture resistance strength at each stage when a static loading force is applied to cause failure; (2) determine the failure mode for each post system at each simulated clinical stage; and (3) determine the feasibility of removing failed post systems. MATERIALS AND METHODS: Ten post systems made with various materials and designs were tested at the following 4 stages of simulated clinical treatment: stage #1: posts only, loaded using a 3-point loading model to failure, to determine transverse strengths and failure modes for each post system; stage #2: posts alone, bonded into teeth; stage #3: posts bonded into teeth with core build up; stage #4: post and core build up and full veneer restoration. For stages #2 through #4, the coronal portion of 60 mandibular premolars was amputated at the cemento-enamel junction, the canals were treated endodontically, and the specimens were mounted in acrylic blocks. A testing force was applied to the posts at 90 degrees to the long axis of the tooth, 4 mm from the cemento-enamel junction. The O'Brien test for constant variance was performed over the treatment groups. For nonconstant variance, the Welsh analysis of variance was used to test for equalities of treatment means. The Tukey Kramer procedure determined which treatment procedures differed. RESULTS: The failure thresholds for each post system were significantly different at each stage of testing, but the order of test results by post type remained generally consistent from one stage to the next. ParaPosts (Coltene, Whaledent Int, New York, NY) and core build up resulted in significantly higher failure thresholds through all 4 stages of testing. This post system also consistently displayed a high number of nonfavorable tooth fractures. FibreKor post and cores (Jeneric Pentron Inc, Wallingford, CT) resulted in significantly lower failure threshold values in stages #2 through #4. This post system displayed no tooth fractures in stages #2 and #3 and a similar number of nonfavorable tooth fractures in stage #4 when compared with the other systems. C-Post (Bisco Dental Products, Schaumburg, IL), CosmoPost (Ivoclar Vivadent North America Inc, Amherst, NY), and AesthetiPost (Bisco Dental Products) grouped in descending order through stages #2 to #4. These systems displayed intermediate fracture resistance strengths, as well as a moderate number of nonfavorable tooth fractures. CosmoPost exhibited a significant number of brittle post fractures with fragments left in the root canal at all stages. The fracture resistance of the cast metal post varied from stage to stage. No teeth fractured at stage #2. At stage #3, 9 of 10 teeth fractured nonfavorably, and all teeth fractured nonfavorably in stage #4. CONCLUSIONS: The fiber posts evaluated provided an advantage over a conventional post that showed a higher number of irretrievable post and unrestorable root fractures. At the stage of final restoration insertion, there was no difference in force to failure for all but the FibreKor material, which continued to be weaker than all other tested materials. The fiber posts were readily retrievable after failure, whereas the remaining post systems tested were nonretrievable. J Prosthodont 2001;10:26-36. PMID- 11406794 TI - Retention of cast posts cemented with zinc phosphate cement using different cementing techniques. AB - PURPOSE: This study compares the effect of different cementation techniques on the retention of cast posts and cores. MATERIALS AND METHODS: Twenty-four extracted single rooted human teeth were used in this study. After routine preparation of the root canals, the canal space was enlarged with reamers. Direct post patterns were made with acrylic resin. Castings were fabricated using Ni-Cr Mb alloy. Zinc phosphate cement was used as a luting agent using 4 different methods of cement introduction to the root canal space. This created 4 treatment groups consisting of 6 teeth in each group. In Group A, the cement was applied over the posts only. For the specimens in Group B, the cement was introduced into the root canals with a spiral filler and applied over the posts. In Group C, cement was injected into the root canals and then applied over the posts. Finally, for the specimens in Group D, cement was injected into the root canals and a spiral was used to spread it into the root canals, and cement was also applied over the posts. After cementation, teeth were stored in saline solution. The tensile retentive force of cast posts was evaluated using the Instron testing machine (Instron, Canton, MA). RESULTS: The retention obtained by the 4 techniques differed significantly (p <.05). The highest tensile force to post dislodgment was observed in Group D, and the least was in Group A. Analysis of variance test revealed a highly significant difference between groups (p <.0001). According to the Fisher exact test, there was a significant increase in retention from Group A to Group D (p <.05). CONCLUSIONS: Under the conditions of this study, the injection of cement into a root canal space followed by the use of a spiral resulted in the highest level of post retention. Retention was reduced when cement was placed using a Lentulo spiral alone (Kerr/Sybron Corp, Romulus, MI), a Jiffy tube (Teledyne Water Pik, Allegheny Teledyne Co, Fort Collins, CO) alone, or by application to the post only. J Prosthodont 2001;10:37-41. PMID- 11406795 TI - Influence of curing method, sealer, and water storage on the hardness of a soft lining material over time. AB - PURPOSE: The purpose of this study was to determine the hardness of plasticized acrylic resin soft lining materials over time when curing procedures were modified and when surface sealers were either used or omitted. MATERIALS AND METHODS: A chairside plasticized acrylic resin material and a heat processed plasticized resin material were made into disks 40 mm in diameter and 10 mm in thickness. Materials were cured by processing at an elevated temperature or at mouth temperature. Five samples were used with each processing method. Half of all samples were treated with a surface sealer, whereas the other samples received no surface treatment. After processing, the samples were immersed in 37 degrees C water for a total time of 1 year. Specimens were tested to determine hardness on a monthly basis. Testing was accomplished using a Shore A durometer. RESULTS: Hardness testing showed differences relative to material and to time after processing. The heat-processed plasticized resin material showed significantly higher Shore A hardness values than the chairside over the entire experimental period (p <.01). Increased hardness was seen for all materials over time, but this was more pronounced with plasticized acrylic resin in which the hardness increased from 28.4% to 115.8% depending on processing method and surface treatment. Surface treatment showed significant softening only when the samples were cured at simulated mouth temperature (p <.05). CONCLUSIONS: Processing method and time after processing have an effect on surface hardness of the tested materials, but neither of these effects is as profound as the initial choice of material. J Prosthodont 2001;10:42-45. PMID- 11406796 TI - The implant-supported milled-bar mandibular overdenture. AB - Osseointegrated dental implants have been proven successful in the treatment of edentulism. The predictability of the implant-supported prosthesis has also been established. Several techniques have been described for the successful restoration of the edentulous mandible: fixed-detachable prostheses with either the original Branemark hybrid prosthesis design or conventional implant-supported fixed partial dentures, implant-retained overdentures, and implant-supported overdentures. However, in cases of advanced ridge resorption in which facial tissue support is needed from the flanges of the prosthesis or when a removable type of prosthesis is preferred by the patient, an implant-supported prosthesis is indicated. Electric discharge machining is often used in the fabrication of the bar for an implant-supported overdenture. This procedure is very costly and technique sensitive. An alternative procedure to fabricate a milled-bar implant supported overdenture is described. This procedure is simple and uses inexpensive equipment and materials. The milled-bar minimizes lateral and rotational displacement. The overdenture incorporates attachments that provide retention, minimizing possible movement along the path of insertion. This type of prosthesis is available to a broad patient population, especially those with advanced ridge resorption, providing an excellent result at a reduced cost. J Prosthodont 2001;10:46-51. PMID- 11406797 TI - The history of articulators: the appearance and early use of the incisal-pin and guide. PMID- 11406798 TI - Qualities of an entrepreneur. PMID- 11406799 TI - The responses of dissociative patients on the thematic apperception test. AB - This study compared the responses of dissociative inpatients and general inpatient psychiatric controls on the Thematic Apperception Test (TAT; Murray, 1943). We found the stories of dissociative participants to be characterized by a greater interpersonal distance and more trauma and dissociation responses than those of the controls. No significant differences were found regarding total number of emotional references, although references to positive emotions were almost nonexistent for the dissociative group. A post hoc analysis of the data found the testing behaviors of dissociative participants to be characterized by switching, trance states, intrainterview amnesias, and affectively loaded card rejections. Questions were raised regarding the relevancy of the findings to clinical practice and how they might explain some of the controversies surrounding the diagnosis of dissociative identity disorder (DID). PMID- 11406800 TI - Syzygy, science, and psychotherapy: the Consumer Reports study. AB - Results from a Consumer Reports (CR) survey indicated that psychotherapy has proven to be quite effective and that longer-term therapy has been more effective than shorter-term therapy. Critiques of the methodology of this study have included the claim that (a) the self-selected sample was biased in favor of people who felt that they had benefited from psychotherapy, (b) the use of retrospective accounts led to a further positive bias, and (c) the validity of the outcome assessment was questionable. Supplemental data from other sources, including prospective data from a large sample of psychotherapy patients, are presented to augment the interpretation of the results of the CR study and to illustrate how some critiques of research results can be evaluated systematically. PMID- 11406801 TI - A survival analysis of clinically significant change in outpatient psychotherapy. AB - The number of sessions required to produce meaningful change has not been assessed adequately, in spite of its relevance to current clinical practice. Seventy-five clients attending outpatient therapy at a university-affiliated clinic were tracked on a weekly basis using the Outcome Questionnaire (Lambert et al., 1996) in order to determine the number of sessions required to attain clinically significant change (CS). Survival analysis indicated that the median time required to attain CS was 11 sessions. When current data were combined with those from an earlier investigation (Kadera, Lambert, and Andrews, 1996), it was found that clients with higher levels of distress took 8 more sessions to reach a 50% CS recovery level than clients entering with lower levels of distress. At a six-month follow-up, CS gains appeared to have been maintained. Other indices of change also were examined (reliable change, average change per session). The implications of these results for allocating mental-health benefits, such as the number of sessions provided through insurance, are discussed. PMID- 11406802 TI - Prediction of dose-response relations based on patient characteristics. AB - The recent discussion of evidence-based, adaptive treatment planning highlights the need for models for the prediction of courses of treatment response. We combine a dose-response model with growth curve modeling to determine dose response relations for well-being, symptoms, and functioning. Hierarchical linear modeling was used to model each patient's expected course of improvement. The resulting predictions were cross-validated on two samples of psychotherapy outpatients. The results give further empirical support for the dose-response model and the phase model of psychotherapy as well as for the usefulness of patient treatment response profiling for individual treatment management. PMID- 11406803 TI - Suicidal ideation and help-negation: Not just hopelessness or prior help. AB - Few distressed young people seek professional psychological help and little is known about what sources of help young people seek for different problems. In suicidal youth, the process of help-negation may exacerbate poor help-seeking. Three hundred and two undergraduate university students completed a questionnaire measuring suicidal ideation, hopelessness, prior help-seeking experience, and help-seeking intentions. Participants indicated they would seek help from different sources of help for different types of problems, but friends consistently were rated as the most likely source of help. Help-negation was suggested by higher levels of suicidal ideation being associated with lower help seeking intentions. However, the negative suicidal ideation/help-seeking intentions relationship was not explained by hopelessness or prior help-seeking. Help-negation appears to involve more than just negative expectations regarding the future. The discussion proposes social problem-solving orientation as one of a number of potential explanatory variables. PMID- 11406804 TI - A factor analytic and psychometric examination of pathology of separation individuation. AB - Two studies are described that attempt to determine if standard-scale-reduction techniques could yield a construct-valid diagnostic screen of pathology of separation-individuation for use in nonclinical university settings. In Study 1 (N = 210), a measure of pathology of separation-individuation (PATHSEP) was reduced successfully to a single, internally consistent factor, accounting for 36% of the variance. In Study 2 (N = 304), these items also coalesced around a single factor, accounting for 35% of the variance. Study 2 also showed that PATHSEP is correlated moderately and positively with indices of insecure attachment, with the Center for Epidemiological Studies-Depression Scale, and with indices of psychiatric symptomatology (Hopkins Symptom Checklist). PATHSEP also was associated with a poorer profile of adjustment to college. Males reported more pathology of separation-individuation than did females. Evidence supports the construct validity of a shortened version of PATHSEP. Directions for future research are noted. PMID- 11406805 TI - Treatment of fibromyalgia incorporating EEG-Driven stimulation: a clinical outcomes study. AB - Thirty patients from a private clinical practice who met the 1990 American College of Rheumatology criteria for fibromyalgia syndrome (FS) were followed prospectively through a brainwave-based intervention known as electroencephalograph (EEG)-driven stimulation or EDS. Patients were initially treated with EDS until they reported noticeable improvements in mental clarity, mood, and sleep. Self-reported pain, then, having changed from vaguely diffuse to more specifically localized, was treated with very modest amounts of physically oriented therapies. Pre- to posttreatment and extended follow-up comparisons of psychological and physical functioning indices, specific FS symptom ratings, and EEG activity revealed statistically significant improvements. EDS appeared to be the prime initiator of therapeutic efficacy. Future research is justified for controlled clinical trials and to better understand disease mechanisms. PMID- 11406806 TI - Relations between personality and depressive symptoms: a multimeasure study of dependency, autonomy, and related constructs. AB - Seventy-four patients seeking treatment for major depression completed four measures of dependent and autonomous personality and two measures of depressive symptomatology. Relationships among the personality measures were investigated by principal-components analysis, enabling systematic comparison of their composition. Relationships between personality components and symptom dimensions were examined to clarify specific associations that have been proposed but inconsistently obtained in previous research. Neither dependency nor autonomy were unitary constructs, and alternative measures had substantial differences in composition. Some support for symptom specificity was obtained. Dependency and autonomy have distinctive associations with depressive symptoms, but their correspondence to unitary personality dimensions and the equivalence of their alternative scales cannot be assumed. PMID- 11406807 TI - Personality characteristics associated with psychological reactance. AB - This study compared six different personality disorders on level of psychological reactance. Eighty clients met criteria for inclusion in one of six personality disorder groups, including passive-aggressive, dependent, personality disorder NOS (not otherwise specified), no personality disorder, obsessive-compulsive, and borderline. Instruments included the Therapeutic Reactance Scale (TRS) and the Questionnaire for the Measurement of Psychological Reactance (QMPR). Four one-way analyses of variance (ANOVAs) were conducted on the scales and subscales of the two reactance measures. All four ANOVAs were significant, although post hoc tests indicated significant differences only among the more extreme groups. The personality disorders were generally ordered on reactance level according to predictions derived from the theory of separation-individuation. PMID- 11406808 TI - Psychometric properties of goal-attainment scaling in the assessment of mann's time-limited psychotherapy. AB - This study tested the reliability and validity of Goal-Attainment Scaling (GAS) in assessing outcome in Mann's Time-Limited Psychotherapy (TLP). Judges constructed GAS scales for 33 patients on five dimensions: severity of symptoms, self-esteem, same-sex friendships, romantic relationships, and work performance. Patients in the treatment group received 12 sessions of TLP, whereas controls were in a waiting list for the same period of time (12 weeks) before starting time-limited psychotherapy. Judges rated all patients on GAS before beginning therapy, at termination, and follow-up. Mean interrater reliability of scores between pairs of judges was r = .88. Convergent validity of the GAS was confirmed by moderate to high correlations with the Health-Sickness Rating Scale (r = .70, p < .001), the Target Complaints Scale (r = .50, p <.01), and the Brief Symptom Inventory (r = .38, p < .05). The findings are discussed and recommendations are made for the reliable and valid use of the GAS in psychotherapy outcome research. PMID- 11406809 TI - No time for complacency: the fetal brain on drugs. PMID- 11406810 TI - Primates exposed to cocaine in utero display reduced density and number of cerebral cortical neurons. AB - This study examined the effects of cocaine use during the second trimester of pregnancy on cerebral neocortical volume and density, and total number of neocortical neurons and glia in offspring. We also evaluated the extent of postnatal recovery of cytoarchitectural abnormalities previously observed in the neocortex of two-month-old primates born from cocaine-treated mothers (Lidow [1995] Synapse 21:332-334). Pregnant monkeys received cocaine orally (20 mg/kg/day) from the 40th to 102nd days of pregnancy (embryonic day [E]40-E102). On E64 and E65, the animals were injected with [(3)H]thymidine. Cerebral hemispheres of the offspring were examined at three years of age. We found a reduction in the neocortical volume and density and total number of neocortical neurons. The observed reduction in neuronal number within the neocortex was not accounted for by the increase in the number of neurons in the white matter of cocaine-exposed animals, because the number of these "extra" neurons was equal to only half that of missing neurons. We detected no significant changes in the number of neocortical glia. The cytoarchitectural abnormalities in the neocortex of prenatally cocaine-exposed three-year-old monkeys closely resembled previously described neocortical abnormalities in similarly exposed two-month-old animals: the neocortex lacked a discernible lamination; the majority of the cells labeled by [(3)H]thymidine injected during neocortical neurogenesis did not reach their proper position within the cortical plate. Therefore, postnatal maturation is not associated with significant improvement in neocortical organization in primates prenatally exposed to cocaine. There was, however, a postnatal recovery of low glial fibrillary acidic protein (GFAP) immunoreactivity previously observed in 2 month-old cocaine-exposed animals. PMID- 11406811 TI - A-fiber sensory input induces neuronal cell death in the dorsal horn of the adult rat spinal cord. AB - Excitotoxicity due to excessive synaptic glutamate release is featured in many neurological conditions in which neuronal death occurs. Whether activation of primary sensory pathways can ever produce sufficient over-activity in secondary sensory neurons in the dorsal horn of the spinal cord to induce cell death, however, has not been determined. In this study, we asked whether activity in myelinated afferents (A fibers), which use glutamate as a transmitter, can induce cell death in the dorsal horn. Using stereological estimates of neuron numbers from electron microscopic sections, we found that stimulation of A-fibers in an intact sciatic nerve at 10 Hz, 20 Hz, and 50 Hz in 10-minute intervals at a stimulus strength that activates both Abeta and Adelta fibers resulted in the loss of 25% of neurons in lamina III, the major site of termination of large Abeta fibers, but not in lamina I, where Adelta fibers terminate. Furthermore, sciatic nerve lesions did not result in detectable neuron loss, but activation of A fibers in a previously sectioned sciatic nerve did cause substantial cell death not only in lamina III but also in laminae I and II. The expansion of the territory of A-fiber afferent-evoked cell death is likely to reflect the sprouting of the fibers into these laminae after peripheral nerve injury. The data show, therefore, that primary afferent A-fiber activity can cause neuronal cell death in the dorsal horn with an anatomical distribution that depends on whether intact or injured fibers are activated. Stimulation-induced cell death potentially may contribute to the development of persistent pain. PMID- 11406812 TI - Dichotomy in phasic-tonic neuromuscular structure of crayfish inhibitory axons. AB - Crustacean muscles are unique in their innervation by both excitatory and inhibitory neurons; therefore, they exhibit polyneuronal and multiterminal innervation. Because excitatory motoneurons are broadly divided into phasic and tonic types, we hypothesized that inhibitory neurons would follow a similar dichotomy. The abdominal extensor muscles in crayfish are separated into parallel deep and superficial bundles; the former has fast muscle fibers innervated by phasic excitatory motoneurons, and the latter has slow fibers supplied by tonic excitatory motoneurons. Each muscle also is innervated by a single, separate inhibitory neuron that uses gamma-aminobutyric acid (GABA) as the inhibitory neurotransmitter. The pattern of axonal branching by the separate inhibitory axons in phasic and tonic abdominal extensor muscles was visualized with confocal microscopy in preparations labeled for GABA-like immunoreactivity. Initial observations indicated that the phasic muscle was covered by extensive GABAergic, filiform axon terminals, whereas innervation of the tonic muscle was comprised of more localized and varicose terminals. With quantitative analyses, we found that the phasic axon has a more highly branched nature than the tonic in first- and second-order branches. The phasic axon branches also were significantly longer than the tonic branches in the second- and third-order branches. Synaptic varicosities in the phasic branches were smaller and less frequent than those in the tonic branches. The fine structure of the inhibitory nerve terminals near synaptic contacts examined with thin-serial-section electron microscopy revealed distinct differences between the phasic system and the tonic system. The phasic terminals were smaller in cross-sectional area than the tonic terminals, and they had smaller synapses and fewer mitochondria. The presynaptic active zone dense bodies were similar in length and number between phasic and tonic synapses. However, their number per synaptic area was two-fold higher in phasic synapses compared with tonic synapses because of the smaller size of the phasic synapses. Thus, within the same neuromuscular system, inhibitory synaptic terminals revealed unique phasic and tonic identities similar to those observed for the excitatory axons. PMID- 11406813 TI - Thalamo-cortical connections of areas 3a and M1 in marmoset monkeys. AB - The present investigation is part of a broader effort to examine cortical areas that contribute to manual dexterity, reaching, and grasping. In this study we examine the thalamic connections of electrophysiologically defined regions in area 3a and architectonically defined primary motor cortex (M1). Our studies demonstrate that area 3a receives input from nuclei associated with the somatosensory system: the superior, inferior, and lateral divisions of the ventral posterior complex (VPs, VPi, and VPl, respectively). Surprisingly, area 3a receives the majority of its input from thalamic nuclei associated with the motor system, posterior division of the ventral lateral nucleus of the thalamus (VL), the mediodorsal nucleus (MD), and intralaminar nuclei including the central lateral nucleus (CL) and the centre median nucleus (CM). In addition, sparse but consistent projections to area 3a are from the anterior pulvinar (Pla). Projections from the thalamus to the cortex immediately rostral to area 3a, in the architectonically defined M1, are predominantly from VL, VA, CL, and MD. There is a conspicuous absence of inputs from the nuclei associated with processing somatic inputs (VP complex). Our results indicate that area 3a is much like a motor area, in part because of its substantial connections with motor nuclei of the thalamus and motor areas of the neocortex (Huffman et al. [2000] Soc. Neurosci. Abstr. 25:1116). The indirect input from the cerebellum and basal ganglia via the ventral lateral nucleus of the thalamus supports its role in proprioception. Furthermore, the presence of input from somatosensory thalamic nuclei suggests that it plays an important role in somatosensory and motor integration. PMID- 11406814 TI - Morphological identification of nitric oxide sources and targets in the cat oculomotor system. AB - Nitric oxide (NO) production by specific neurons in the prepositus hypoglossi (PH) nucleus is necessary for the correct performance of eye movements in alert cats. In an attempt to characterize the morphological substrate of this NO function, the distribution of nitrergic neurons and NO-responding neurons has been investigated in different brainstem structures related to eye movements. Nitrergic neurons were stained by either immunohistochemistry for NO synthase I or histochemistry for reduced nicotinamide adenine dinucleotide phosphate (NADPH) diaphorase. The NO targets were identified by cyclic guanosine monophosphate (cGMP) immunohistochemistry in animals treated with a NO donor immediately before fixation of the brain. Connectivity between cells of the NO-cGMP pathway was analyzed by injections of the retrograde tracers horseradish peroxidase or fast blue in different structures. The motor nuclei commanding extraocular muscles did not contain elements of the NO-cGMP pathway, except for some scattered nitrergic neurons in the most caudal part of the abducens nucleus. The PH nucleus contained the largest number of nitrergic cell bodies and a rich neuropil, distributed in two groups in medial and lateral positions in the caudal part, and one central group in the rostral part of the nucleus. An abundant cGMP positive neuropil was the only NO-sensitive element in the PH nucleus, where no cGMP-producing neuronal cell bodies were observed. The opposite disposition was found in the marginal zone between the PH and the medial vestibular nuclei, with a large number of NO sensitive cGMP-producing neurons and almost no nitrergic cells. Both nitrergic and NO-sensitive cell bodies were found in the medial and inferior vestibular nuclei and in the superior colliculus, whereas the lateral geniculate nucleus contained nitrergic neuropil and a large number of NO-sensitive cell bodies. Some of the cGMP-positive neurons in the marginal zone and medial vestibular nucleus projected to the PH nucleus, predominantly to the ipsilateral side. These morphological findings may help to explain the mechanism of action of NO in the oculomotor system. PMID- 11406815 TI - Topographic organization and neurochemical identity of dorsal raphe neurons that project to the trigeminal somatosensory pathway in the rat. AB - The primary goals of this study were to: 1) examine the distribution of neurons within the dorsal raphe (DR) nucleus that project to cortical and subcortical sites along the trigeminal somatosensory pathway in rat; 2) determine the extent to which different regions within this ascending sensory system receive collateral projections from the same DR neuron; and 3) identify the putative transmitters contained within these DR projection neurons. Long-Evans hooded rats received pressure injections of various combinations of retrograde fluorescent tracers; into the whisker-related regions of the primary somatosensory cortex (barrel field cortex [BC]), ventral posterior medial thalamus (VPM), and principal nucleus of the trigeminal complex (PrV). The distribution of retrogradely labeled neurons within the DR was examined by fluorescence microscopy. The major finding was that cortically projecting neurons were located within the midline regions of the rostral portion of the DR, whereas cells projecting to subcortical trigeminal somatosensory structures were distributed bilaterally in the lateral wing regions of the DR as well as in the midline portions of the nucleus. Single neurons that send axon collaterals to multiple cortical and subcortical trigeminal somatosensory targets were observed in the dorsomedian and ventromedian regions of the DR. DR neurons that projected to cortical and subcortical sites contained serotonin but not tyrosine hydroxylase, the marker enzyme for catecholamine transmitters. Taken together, these findings provide further evidence of neurochemical specificity and functional anatomical organization within the DR efferent projection system. PMID- 11406816 TI - Jaw-muscle spindle afferent pathways to the trigeminal motor nucleus in the rat. AB - Neural pathways conveying proprioceptive feedback from the jaw muscles were studied in rats by combining retrograde and intracellular neuronal labeling. Initially, horseradish peroxidase was iontophoresed unilaterally into the trigeminal motor nucleus (Vmo). Two days later, 1-5 jaw-muscle spindle afferent axons located in the mesencephalic trigeminal nucleus were physiologically identified and intracellularly stained with biotinamide. Stained mesencephalic trigeminal jaw-muscle spindle afferent axon collaterals and boutons were predominantly distributed in the supratrigeminal region (Vsup), Vmo, dorsomedial trigeminal principal sensory nucleus (Vpdm), parvicellular reticular formation (PCRt), alpha division of the parvicellular reticular formation (PCRtA), and dorsomedial portions of the spinal trigeminal subnuclei oralis (Vodm), and interpolaris (Vidm). Numerous neurons retrogradely labeled with horseradish peroxidase from the trigeminal motor nucleus were found bilaterally in the PCRt, PCRtA, Vodm, and Vidm. Retrogradely labeled neurons were also present contralaterally in the Vsup, Vpdm, Vmo, peritrigeminal zone, and bilaterally in the dorsal medullary reticular field. Putative contacts between intracellularly stained mesencephalic trigeminal jaw-muscle spindle afferent boutons and trigeminal premotor neurons retrogradely labeled with horseradish peroxidase were found in the ipsilateral Vodm, PCRtA, and PCRt, as well as the contralateral Vsup, Vmo, Vodm, PCRt, and PCRtA. Thus, multiple disynaptic jaw-muscle spindle afferent-motoneuron circuits exist. These pathways are likely to convey long latency jaw-muscle stretch reflexes and may contribute to stiffness regulation of the masticatory muscles. PMID- 11406817 TI - SPACRCAN in the developing retina and pineal gland of the rat: spatial and temporal pattern of gene expression and protein synthesis. AB - SPACRCAN is a hyaluronan-binding proteoglycan that is present in the pineal gland and interphotoreceptor matrix of the retina. Here, we evaluate the pattern of SPACRCAN gene expression and protein appearance during retinal and pineal gland development in the rat. In situ hybridization histochemistry with SPACRCAN riboprobes indicates that hybridization signals are first evident in the retina over developing photoreceptor cells at embryonic day 16 (E16) and in the pineal gland at E21. Immunocytochemistry using a SPACRCAN antibody shows localization of SPACRCAN protein in the developing interphotoreceptor matrix by Postnatal day 5 (P5) and in the pineal gland by P6. These studies suggest that SPACRCAN mRNA expression may occur substantially earlier than the time when SPACRCAN protein is detectable in both the retina and the pineal gland. The period of retinal histogenesis when SPACRCAN is detected first is coincident with the time photoreceptors begin to extend from the outer retinal surface, suggesting that SPACRCAN may participate in the maturation and maintenance of the light-sensitive photoreceptor outer segment. PMID- 11406818 TI - Distribution of nonmuscle myosin-II in honeybee photoreceptors and its possible role in maintaining compound eye architecture. AB - Photoreceptor and accessory cells in the insect compound eye exhibit a characteristic architecture, probably established and maintained by the contribution of membrane-associated cytoskeletal elements. The present study identifies and localizes nonmuscle myosin-II in honeybee photoreceptors by use of an affinity-purified antibody against scallop muscle myosin-II heavy chain (MHC). Western blot analysis and immunofluorescence staining confirmed cross-reactivity of the antibody with honeybee muscle MHC. In the compound eye, the antibody identified a protein that comigrated with muscle MHC on sodium dodecylsulfate polyacrylamide gels. Association with the cytoskeleton, ATP-dependent binding to exogenous actin filaments, and cross-reactivity with several other antibodies against MHC, including an antibody to Drosophila nonmuscle MHC, support the conclusion that the cross-reacting protein represents nonmuscle MHC. Confocal immunofluorescence microscopy on honeybee eyes showed that the motor protein was highly enriched at distinct regions of the photoreceptor surface next to the light-receptive compartment, the rhabdom. To determine the function of myosin-II in these cells, retinal tissue was incubated with 2,3-butanedione 2-monoxime (BDM), an inhibitor of myosin activity. BDM treatment resulted in an increase in surface curvature at precisely those membrane areas that exhibited intense immunoreactivity for MHC. Moreover, the positioning and alignment of the rhabdoms was altered after exposure to BDM. These results suggest that the activity of nonmuscle myosin-II in the visual cells exerts tension on a distinct surface region next to the rhabdom, contributes to the positioning of the rhabdom, and, thus, plays a role in maintaining the cellular architecture within the compound eye. PMID- 11406819 TI - Immunocytochemical localization of candidates for vesicular glutamate transporters in the rat cerebral cortex. AB - Brain-specific Na(+)-dependent inorganic phosphate cotransporter (BNPI) was recently reported to serve as a vesicular glutamate transporter (VGluT), and was renamed VGluT1 (Bellocchio et al. [ 2000] Science 289:957-960; Takamori et al. [2000] Nature 407:189-194). Ahead of these reports, cDNA encoding another brain specific inorganic phosphate transporter, which showed 82% amino acid identity to VGluT1, was cloned and designated differentiation-associated Na(+)-dependent inorganic phosphate cotransporter (DNPI; Aihara et al. [2000] J Neurochem 74:2622 2625). In the present study, we produced a specific antibody against a C-terminal portion of DNPI, and studied the immunohistochemical localization of DNPI in the rat cerebral cortex in comparison with that of VGluT1. DNPI immunoreactivity was enriched in neuropil of layers I and IV and to a lesser extent in the upper portion of layer VI of the cerebral neocortex, whereas VGluT1 immunoreactivity was distributed more evenly in neuropil of the neocortex. Electron microscopic observation revealed that both DNPI and VGluT1 immunoreactivities were mainly located on synaptic vesicles in nerve terminals which made asymmetrical contacts in the neocortex. Furthermore, neither DNPI nor VGluT1 immunoreactivity in the neocortex was colocalized with gamma aminobutyric acid (GABA)ergic axon terminal markers, immunoreactivity for glutamic acid decarboxylase or vesicular GABA transporter. Neuronal depletion in the ventrobasal thalamic nuclei produced by the kainic acid injection resulted in a clear reduction of DNPI immunoreactivity in layers I, IV, and VI of the somatosensory cortex. These results indicate that DNPI is located on the membrane of synaptic vesicles in thalamocortical axon terminals, and that it may be a candidate for VGluT of thalamocortical glutamatergic neurons. PMID- 11406820 TI - Lancelet lessons: evaluating a phylogenetic model. PMID- 11406821 TI - Establishment of left-right asymmetric innervation in the lancelet oral region. AB - Lancelets (amphioxus) exhibit a remarkable asymmetric development in the anterior body region, which is reflected in the peripheral nervous system even at adulthood. Not all of the anterior nerves are involved, but the left third to fifth nerves are clearly asymmetric. To trace the developmental process responsible for asymmetric innervation, the peripheral nerves in the anterior region were studied in pre- and mid-metamorphic larvae, 1-cm-long juveniles, and in adults by using whole-mount immunostaining. The mouth changes in size and location during larval life before moving ventrally and, in conjunction with this change, nerves in the oral region are also modified. The left second nerve initially innervates the oral region, but this connection is secondarily lost. As the mouth expands and shifts posteriorly, the left fifth to ninth nerves join the left third and fourth in the innervation of the oral region. The left third to sixth nerves anastomose with the oral nerve ring, which encircles the mouth on the left side. In the juveniles and adults, there are two nerve plexuses that run parallel to the margin of the oral hood. The innermost of these, the "inner oral hood nerve plexus", is asymmetrically connected with the left third to fifth nerves on both sides. The other, the "outer oral-hood nerve plexus", is ipsilaterally connected with the third to seventh nerves on both sides. The velar nerve ring is also innervated asymmetrically by the left fourth and fifth nerves. From these observations, we suggest that the oral nerve ring is the precursor of both the inner oral-hood nerve plexus and the velar nerve ring, and that the asymmetric innervation retained in adult lancelets is related to the early anastomosis of the left nerves with the oral nerve ring. We also show that, contrary to the persistent asymmetric innervation, the axonal patterns of the anterior peripheral nervous system in developing lancelets can change. PMID- 11406822 TI - Adult neurogenesis produces a large pool of new granule cells in the dentate gyrus. AB - Knowing the rate of addition of new granule cells to the adult dentate gyrus is critical to understanding the function of adult neurogenesis. Despite the large number of studies of neurogenesis in the adult dentate gyrus, basic questions about the magnitude of this phenomenon have never been addressed. The S-phase marker bromodeoxyuridine (BrdU) has been extensively used in recent studies of adult neurogenesis, but it has been carefully tested only in the embryonic brain. Here, we show that a high dose of BrdU (300 mg/kg) is a specific, quantitative, and nontoxic marker of dividing cells in the adult rat dentate gyrus, whereas lower doses label only a fraction of the S-phase cells. By using this high dose of BrdU along with a second S-phase marker, [(3)H]thymidine, we found that young adult rats have 9,400 dividing cells proliferating with a cell cycle time of 25 hours, which would generate 9,000 new cells each day, or more than 250,000 per month. Within 5-12 days of BrdU injection, a substantial pool of immature granule neurons, 50% of all BrdU-labeled cells in the dentate gyrus, could be identified with neuron-specific antibodies TuJ1 and TUC-4. This number of new granule neurons generated each month is 6% of the total size of the granule cell population and 30-60% of the size of the afferent and efferent populations (West et al. [1991] Anat Rec 231:482-497; Mulders et al. [1997] J Comp Neurol 385:83 94). The large number of the adult-generated granule cells supports the idea that these new neurons play an important role in hippocampal function. PMID- 11406823 TI - Spatial arrangement of cerebro-pontine terminals. AB - Understanding the interaction of the cerebral cortex and cerebellum requires knowledge of the highly complex spatial characteristics of cerebro-cerebellar signal transfer. Cerebro-pontine fibers from one neocortical site terminate in several sharply demarcated patches across large parts of the pontine nuclei (PN), and fibers from different neocortical areas terminate in the same pontine region. To determine whether projections from segregated neocortical sites overlap in the PN, we studied double anterograde tracing of cerebro-pontine terminals from large parts of rat neocortex. In none of these experiments, including double injection into two functionally related areas, were we able to demonstrate overlapping patches, although close spatial relationships were always detected. This non overlapping distribution is consistent with a compartmentalized organization of the cerebro-pontine projection and may be the basis of the fractured type of maps found in the cerebellar granular layer. The critical distance between two sites on the neocortical surface that project to non-overlapping patches in the PN was found to be 600 microm, by using double injection within the whisker representation of the primary somatosensory area. This matches the diameter of dendritic trees of layer 5 projection neurons, indicating that non-overlapping populations of neocortical projection neurons possess non-overlapping patches of pontine terminals. Estimations based on this critical distance and the pontine volume anterogradely labeled by one injection site indicate that the size of the PN may be well suited to accommodate a complete set of non-overlapping pontine patches from all possible neocortical sites. PMID- 11406824 TI - Selective synaptic distribution of AMPA and kainate receptor subunits in the outer plexiform layer of the carp retina. AB - The subunit composition of ionotropic glutamate receptors (GluRs) is extremely diverse and responsible for the diversity of postsynaptic responses to the release of glutamate, which is the major excitatory neurotransmitter in the retina. To understand the functional consequences of this diversity, it is necessary to reveal the synaptic localization and subunit composition of GluRs. We have used immuno light and electron microscopy to localize AMPA and kainate (GluR1, GluR2/3, GluR4, GluR5-7) subunits in identified carp retinal neurons contributing to the outer plexiform layer. GluR1 could not be detected within the outer plexiform layer. Rod and cone horizontal cells all express only GluR2/3 at the tips of their invaginating dendrites. These receptors are also inserted into the membrane of spinules, light-dependent protrusions of the horizontal cell dendrites, flanking the synaptic ribbon of the cone synapse. Bipolar cells express GluR2/3, GluR4, and GluR5-7 at their terminal dendrites invaginating cone pedicles and rod spherules. Colocalization data suggest that each subunit is expressed by a distinct bipolar cell type. The majority of bipolar cells expressing these receptors seem to be of the functional OFF-type; however, in a few instances, GluR2/3 could also be detected on dendrites of bipolar cells that, based on their localization within the cone synaptic complex, appeared to be of the functional ON-type. The spatial arrangement of the different subunits within the cavity of the cone pedicle appeared not to be random: GluR2/3 was found predominantly at the apex of the cavity, GluR4 at its base and GluR5-7 dispersed between the two. PMID- 11406825 TI - Evidence for regional heterogeneity in corticotropin-releasing factor interactions in the dorsal raphe nucleus. AB - The dorsal raphe nucleus (DR) is innervated by fibers containing the stress related neurohormone corticotropin-releasing factor (CRF), which alters DR neuronal activity and serotonin release in rats. This study examined the relative distribution of CRF-immunoreactive fibers in the rat DR by using light level densitometry. Additionally, CRF-immunoreactive processes within specific subregions of the DR were examined at the ultrastructural level by using electron microscopy. CRF-immunoreactive fibers were organized within the DR along a caudal rostral gradient, such that proceeding rostrally, innervation shifted from dorsolateral to ventromedial. Numerous CRF-immunoreactive axon terminals containing dense-core vesicles were found in both the caudal dorsolateral region and the rostral ventromedial/interfascicular region. These formed synaptic specializations with unlabeled dendrites and frequently contacted nonlabeled axon terminals. Semiquantitative analysis revealed certain differences between the two regions with respect to the types of associations made by CRF-immunoreactive terminals. Associations with dendrites were more frequent in the dorsolateral vs. ventromedial region (65% of 171 terminals vs. 39% of 233 terminals, respectively), whereas associations with axon terminals were more frequent in the ventromedial/interfascicular vs. the dorsolateral region (72% of 233 terminals vs. 57% of 171 terminals, respectively). Additionally, synaptic specializations between CRF-immunoreactive terminals and dendrites were more frequently asymmetric in the dorsolateral region (60%) and symmetric (49%) in the ventromedial/interfascicular region. Regional differences in CRF terminal interactions in the DR could account for the reported heterogeneous effects of CRF on DR neuronal activity and forebrain serotonin release. Importantly, the present results provide anatomical substrates for regulation of the DR by endogenous CRF. PMID- 11406826 TI - Developmental regulation of CPG15 expression in Xenopus. AB - Mechanisms controlling dendritic arbor formation affect the establishment of neuronal circuits. Candidate plasticity gene 15 (CPG15) is a glycosylphosphatidyl inositol (GPI)-linked activity-induced protein that has been shown to function as an intercellular signaling molecule that can promote the morphological and physiological development of the Xenopus retinotectal system. A thorough understanding of CPG15 function requires knowledge of the spatiotemporal expression of the endogenous protein. We therefore cloned Xenopus cpg15 and used RNA in situ hybridization and immunohistochemistry to determine the pattern of CPG15 expression. cpg15 mRNA and CPG15 protein are first detectable in the developing spinal cord and become widespread as development proceeds. CPG15 is expressed in sensory regions of the brain, including the visual, auditory, and olfactory systems. Within the retina, CPG15 is only expressed in retinal ganglion cells. CPG15 protein is concentrated in axon tracts, including retinal axons. These data support a model in which CPG15 expressed in retinal ganglion cells is trafficked to retinal axons, where it modulates postsynaptic dendritic arbor elaboration, and synaptic maturation. PMID- 11406827 TI - Subdivisions of hymenopteran mushroom body calyces by their afferent supply. AB - The mushroom bodies are regions in the insect brain involved in processing complex multimodal information. They are composed of many parallel sets of intrinsic neurons that receive input from and transfer output to extrinsic neurons that connect the mushroom bodies with the surrounding neuropils. Mushroom bodies are particularly large in social Hymenoptera and are thought to be involved in the control of conspicuous orientation, learning, and memory capabilities of these insects. The present account compares the organization of sensory input to the mushroom body's calyx in different Hymenoptera. Tracer and conventional neuronal staining procedures reveal the following anatomic characteristics: The calyx comprises three subdivisions, the lip, collar, and basal ring. The lip receives antennal lobe afferents, and these olfactory input neurons can terminate in two or more segregated zones within the lip. The collar receives visual afferents that are bilateral with equal representation of both eyes in each calyx. Visual inputs provide two to three layers of processes in the collar subdivision. The basal ring is subdivided into two modality-specific zones, one receiving visual, the other antennal lobe input. Some overlap of modality exists between calycal subdivisions and within the basal ring, and the degree of segregation of sensory input within the calyx is species-specific. The data suggest that the many parallel channels of intrinsic neurons may each process different aspects of sensory input information. PMID- 11406828 TI - Intrinsic connectivity of the rat subiculum: I. Dendritic morphology and patterns of axonal arborization by pyramidal neurons. AB - The dendritic and axonal morphology of rat subicular neurons was studied in single cells labeled with Neurobiotin. Electrophysiological classification of cells as intrinsic burst firing or regular spiking neurons was correlated with morphologic patterns and cell locations. Every cell had dendritic branches that reached the outer molecular layer, with most cells having branches that reached the hippocampal fissure. All but two pyramidal cells had axon collaterals that entered the deep white matter (alveus). Branching patterns of apical dendrites varied as a function of the cell's soma location along the fissure-alveus axis of the cell layer. The first major dendritic branch point for most cells occurred at the superficial edge of the cell layer giving deep cells long primary apical dendrites and superficial cells short or absent primary apical dendrites. In contrast, basal dendritic arbors were similar across cells regardless of cell position. Apical and basal dendrites of all cells had numerous spines. Superficial and deep cells also differed in axonal collateralization. Deep cells (mostly intrinsically bursting [IB] class) had one or more ascending axon collaterals that typically remained within the region circumscribed by their apical dendrites. Superficial cells (mostly regular spiking [RS] class) tended to have axon collaterals that reached longer distances in the cell layer. Numerous varicosities and axonal extensions were present on axon collaterals in the cell layer and in the apical dendritic region, suggesting intrinsic connectivity. Axonal varicosities and extensions were found on axons that entered presubiculum, entorhinal cortex or CA1, supporting the notion that these were projection cells. Local collaterals were distinctly thinner than collaterals that would leave the subiculum, suggesting little or no myelin on local collaterals and some myelin on efferent fibers. We conclude that both IB and RS classes of subicular principal cells make synaptic contacts in and apical to the cell layer. Based on the patterns of axonal arborization, we suggest that subiculum has at least a crude columnar and laminar architecture, with ascending collaterals of deep cells forming columns and broader axonal arbors of superficial cells serving to distribute activity across multiple columns. PMID- 11406829 TI - Intrinsic connectivity of the rat subiculum: II. Properties of synchronous spontaneous activity and a demonstration of multiple generator regions. AB - Brain structures that can generate epileptiform activity possess excitatory interconnections among principal cells and a subset of these neurons that can be spontaneously active ("pacemaker" cells). We describe electrophysiological evidence for excitatory interactions among rat subicular neurons. Subiculum was isolated from presubiculum, CA1, and entorhinal cortex in ventral horizontal slices. Nominally zero magnesium perfusate, picrotoxin (100 microM), or NMDA (20 microM) was used to induce spontaneous firing in subicular neurons. Synchronous population activity and the spread of population events from one end of subiculum to the other in isolated subicular subslices indicate that subicular pyramidal neurons are coupled together by excitatory synapses. Both electrophysiological classes of subicular pyramidal cells (bursting and regular spiking) exhibited synchronous activity, indicating that both cell classes are targets of local excitatory inputs. Burst firing neurons were active in the absence of synchronous activity in field recordings, indicating that these cells may serve as pacemaker neurons for the generation of epileptiform activity in subiculum. Epileptiform events could originate at either proximal or distal segments of the subiculum from ventral horizontal slices. In some slices, events originated in both proximal and distal locations and propagated to the other location. Finally, propagation was supported over axonal paths through the cell layer and in the apical dendritic zone. We conclude that subicular burst firing and regular spiking neurons are coupled by means of glutamatergic synapses. These connections may serve to distribute activity driven by topographically organized inputs and to synchronize subicular cell activity. PMID- 11406831 TI - Cytotoxic and genotoxic effects of As, MMA, and DMA on leukocytes and stimulated human lymphocytes. AB - Inorganic arsenic is a human carcinogen associated with different types of cancer. Arsenic metabolism produces two methylated species: monomethylarsonic and dimethylarsinic acids. Although this metabolic route has been involved in arsenic detoxification, it is still not clear whether these methylated metabolites participate in the carcinogenic process. In this work, we studied the cytotoxic and genotoxic effects of arsenic and its metabolites. Cytotoxicity was evaluated in cultured lymphocytes from three donors. Mitotic and replication indices were the parameters analyzed. The results indicate a clear cytotoxic effect by sodium arsenite but not by its metabolites. Genotoxicity was assessed by the single cell gel electrophoresis assay. Sodium arsenite increased DNA migration in stimulated lymphocytes only at doses greater than 5 x 10(-6) M; meanwhile in leukocytes a weak response was observed. Monomethylarsonic acid produced in leukocytes a weak induction of DNA damage, while in stimulated lymphocytes, a dose-increase in DNA migration was observed. The injury caused by dimethylarsinic acid was more evident than that observed in cultures treated with sodium arsenite and monomethylarsonic acid in stimulated lymphocytes, although in leukocytes no effect on DNA migration was found. In conclusion, only sodium arsenite had the capacity to alter mitotic and replication indices, while sodium arsenite and its metabolites were capable of inducing single strand DNA breaks on stimulated human lymphocytes treated in vitro for 24 h; however, the differences observed were between individual responses, one donor being more susceptible even at the lower doses. This individual susceptibility to arsenic compounds has been repeatedly observed for different end-points and should be studied further. PMID- 11406832 TI - Examination of Alternaria alternata mutagenicity and effects of nitrosylation using the Ames Salmonella test. AB - Molds of the genus Alternaria are common food pathogens responsible for the spoilage of fruits, vegetables, grains, and nuts. Although consumption of Alternaria alternata-contaminated foodstuffs has been implicated in an elevated incidence of esophageal carcinogenesis, the mutagenic potencies of several A. alternata toxins seem unable to account for the levels of activity found using crude mycelial extracts. In this study, the mutagenic effects of nitrosylation were examined with the major Alternaria metabolites Altenuene (ALT), Alternariol (AOH), Alternariol Monomethyl Ether (AME), Altertoxin I (ATX I), Tentoxin (TENT), Tenuazonic Acid (TA), and Radicinin (RAD) using the Ames Salmonella strains TA98 and TA100. In the absence of nitrosylation, ATX I was mutagenic when tested from 1 to 100 microg/plate in TA98 with rat liver S9 for activation, while AOH and ATX I were weakly mutagenic +/- S9 in TA100. Incubation with nitrite generally increased mutagenic potencies with ATX I strongly mutagenic +/- S9 in both TA98 and TA100, while ALT, AOH, AME, and RAD responses were enhanced in TA100 + S9. However, subsequent examination of three extracts made from A. alternata culture broth, acetone-washed mycelia, and the acetone washes showed a different mutagenic response with both broth and acetone washes directly mutagenic in TA98 and TA100 but with a reduced response + S9. The acetone-washed mycelial extract was found to have the lowest mutagenic activity of the three extracts tested. Nitrosylation had little effect on the mutagenicity of any of the extracts. Thus, while nitrosylation increases the mutagenicity of ATX I, and to a lesser extent that of several other Alternaria toxins, the results demonstrate that Alternaria produces a major mutagenic activity with a S. typhimurium response different from that found with the purified toxins. Efforts are currently underway to chemically identify this mutagenic species. Published 2001 Wiley-Liss, Inc. PMID- 11406833 TI - Characterization of the mutational specificity of DNA cross-linking mutagens by the Lac+ reversion assay with Escherichia coli. AB - The mutational specificities of DNA cross-linking compounds such as cisplatin, transplatin, carboplatin, mitomycin C, psoralen, and 8-methoxypsoralen were investigated in lacZ reversion assay systems of Escherichia coli. Tester strains were constructed by introducing the six kinds of F' plasmids (lacI-, lacZ461, and proAB+), each of which carries a different base-substitution mutation within the lacZ gene. Each of the six possible base-substitution mutations was assayed by Lac+ reversion. Cisplatin induced G.C-->A.T transitions and G.C-->T.A transversions, with the former predominating. Transplatin induced A.T-->G.C transitions in addition to G.C-->A.T transitions and G.C-->T.A. Carboplatin weakly induced G.C-->A.T transitions. On the other hand, mitomycin C induced only G.C-->T.A transversions, while psoralen and 8-methoxypsoralen reactivated with near-UV irradiation induced A.T-->G.C transitions preferentially. The Lac(+) reversion system was very convenient for rapidly determining mutational spectra. PMID- 11406834 TI - Teratogenic effects of Doxorubicin in rats at midgestation and at term. AB - Doxorubicin (DOXO) is an anticancer drug widely used against leukaemias and solid tumours. Doxorubicin is teratogenic in laboratory species, as demonstrated by a number of in vivo and in vitro experiments. In our in vitro study, the endoderm of the primitive gut and the caudal mesenchymes have been the main target tissues, while oxidative stress has been excluded as a mechanism in Doxorubicin related abnormalities. The aims of the present work are the confirmation of our previous in vitro results (obtained on rat embryos cultured from 9.5 to 11.5 d.p.c.) in an in vivo model (midgestation) and the comparison of the abnormalities observed in the embryos at midgestation with malformations observable at term of pregnancy. For this purpose, pregnant rats were i.p. treated on 9.5 and 10.5 d.p.c. with DOXO. Embryos were analysed on 11 d.p.c., while foetuses were externally and viscerally analysed on 20 d.p.c. (term of gestation). The data collected at midgestation confirm the specific embryotoxic effects (particularly to the primitive gut and the caudal mesenchyme) previously observed in vitro. The data collected at term show the endoderm derivatives (gastrointestinal apparatus and respiratory tract) as the primary target organs, supporting the idea that the teratogenic activity of Doxorubicin is mediated by specific toxic effects directed to the primitive gut. PMID- 11406835 TI - Promotion of chemically induced rat esophageal tumorigenesis with post-initiation ethanol modification. AB - Post-initiation ethanol modification on N-nitrosomethylbenzylamine (NMBA)-induced rat esophageal carcinogenesis model was investigated in male, 6-week-old, F344 rats that received s.c. injections, 3 times per week, of 0.5 mg/kg NMBA for the first 5 weeks and then were treated with 0% (Group 1), 3.3% (Group 2), and 10% (Group 3) ethanol in the drinking water for up to 20 weeks. Group 4 received 10% ethanol without NMBA administration and Group 5 was maintained without any chemical treatment. There were no statistical differences in the incidence and multiplicity of esophageal tumors among Groups 1 to 3. However, the multiplicity of hyperplasias was statistically greater in Group 3 than in Groups 1 or 2. Esophageal epithelia of all rats in Groups 4 and 5 demonstrated a normal histology. BrdU labelling indices of tumors and hyperplasias in NMBA-treated groups were essentially similar, although cycline D1 was overexpressed to a greater extent in tumors and also hyperplasias of Group 3 than in Groups 1 or 2. The results indicated ethanol to exert weak promotion effects through cycline D1 overexpression on rat esophageal tumorigenesis initiated with NMBA. PMID- 11406836 TI - Protective effect of Picroliv, the active constituent of Picrorhiza kurroa, against chemical carcinogenesis in mice. AB - Cancer chemoprevention of chemically induced tumours by Picroliv, an iridoid glycoside mixture purified from Picrorhiza kurroa, was studied on 20 methylcholanthrene (20-MC)-induced sarcoma model and 7,12 dimethylbenz[a]anthracene (DMBA)-initiated papilloma formation in BALB/c mice. Administration of Picroliv (100 and 200 mg/kg, p.o) inhibited the sarcoma development by 47 and 53% as estimated on day 200 after 20-MC administration. Control animals started dying of tumour burden 76 days after 20-MC administration and all animals were dead by day 170, while 60 and 66% of the animals survived in the Picroliv treated group, 100 and 200 mg/kg, respectively. Picroliv exhibited anti-tumour-promoting activity on a two-stage carcinogenesis test on mouse skin using DMBA as an initiator and croton oil as a promoter. Topical application of Picroliv (1 and 5 mg/mouse) 30 minutes prior to that of croton oil application resulted in a 50 and 60% reduction in the number of animals that developed papillomas, and 48 and 64% reduction in the number of papillomas per mouse. There was also a delay in the onset of first skin tumour in the group of animals treated with Picroliv. Oral administration of Picroliv (150 mg/kg, p.o.) prior to DMBA application delayed the onset of papillomas and the percent of mice (60%) with tumours indicates that Picroliv inhibited the tumour initiation induced by DMBA. Picroliv administration was also found to increase the life span of transplanted Dalton's Lymphoma Ascites (DLA) and Ehrlich Ascites Carcinoma (EAC) harboring mice and reduced the volume of transplanted solid tumours. PMID- 11406837 TI - Randomization at the level of primary care practice: use of pre-intervention data and random effects models. AB - The Anglia menorrhagia education study tests the effectiveness of an education package for the treatment of menorrhagia given to doctors at a primary care level. General practices were randomized to receive or not receive the package. It is hoped that this intervention will reduce the proportion of women suffering from menorrhagia that are referred to hospital. Data are available on the treatment and referral of women in the practices in the education and control groups, both pre- and post-intervention. We define and demonstrate a random effects logistic regression model that includes pre-intervention data for calculating the effectiveness of the intervention. PMID- 11406838 TI - Estimating probability of non-response to treatment using mixture distributions. AB - Repeat measurements of patient characteristics are often used to assess response to treatment. In this paper we discuss a normal mixture model for the observed change in the characteristic of interest in treated patients. The methods described can be used to estimate the overall proportion of non-response to treatment and also the probability that a patient has not responded to treatment given his or her observed change. The model parameters are estimated using maximum likelihood, and the delta method is used to construct a pointwise confidence band for the conditional probability that a patient is a non-responder to treatment. The work was initially motivated by analysis issues in the Fracture Intervention Trial (FIT), a randomized trial of the osteoporosis drug alendronate, and the method is illustrated with data from that study. We also evaluate key aspects of the estimation procedure with two simulation studies. In the first, the data generation model is the assumed normal mixture model, and in the second, the data are generated according to a shifted and scaled central t distribution model suggested by the FIT data. PMID- 11406839 TI - A multiple imputation method for missing covariates in non-linear mixed-effects models with application to HIV dynamics. AB - We propose a three-step multiple imputation method, implemented by Gibbs sampler, for estimating parameters in non-linear mixed-effects models with missing covariates. Estimates obtained by the proposed multiple imputation method are compared to those obtained by the mean-value imputation method and the complete case method through simulations. We find that the proposed multiple imputation method offers smaller biases and smaller mean-squared errors for the estimates of covariate coefficients compared to other two methods. We apply the three missing data methods to modelling HIV viral dynamics from an AIDS clinical trial. We believe that the results from the proposed multiple imputation method are more reliable than that from the other two commonly used methods. PMID- 11406840 TI - On tests of the overall treatment effect in meta-analysis with normally distributed responses. AB - For the meta-analysis of controlled clinical trials or epidemiological studies, in which the responses are at least approximately normally distributed, a refined test for the hypothesis of no overall treatment effect is proposed. The test statistic is based on a direct estimation function for the variance of the overall treatment effect estimator. As outcome measures, the absolute and the standardized difference between means are considered. In simulation studies it is shown that the proposed test keeps the prescribed significance level very well in contrast to the commonly used tests in the fixed effects and random effects model, respectively, which can become very liberal. Furthermore, just for using the proposed test it is not necessary to choose between the fixed effects and the random effects approach in advance. PMID- 11406841 TI - Testing for the presence of cured patients: a simulation study. AB - An important but difficult problem in clinical trials is to determine the presence of cured patients when long-term survivors are observed. The likelihood ratio test has been studied for this purpose in the gamma mixture model. However, its asymptotic null distribution is not readily available due to a violation of boundary conditions in the standard asymptotic theory. In this paper, a simulation study is employed to examine a proposed asymptotic null distribution of the likelihood ratio test. We find that the distribution can also be used to approximate the asymptotic null distribution of the likelihood ratio test in the Weibull and log-normal mixture models when the censoring rate is not too light. However, the simulation study also shows that null distribution of the likelihood ratio test deviates significantly from the suggested distribution under moderate sample sizes when the censoring rate is small or the hazard rate is large. Consequently caution is needed in this case to determine the presence of cured patients. Finally, the results are used to confirm the presence of cured patients in a leukaemia study. PMID- 11406842 TI - Notes on interval estimation of the attributable risk in cross-sectional sampling. AB - The attributable risk (AR) is probably the most useful and commonly used epidemiologic index to measure the importance of a risk factor in public health issues. This paper focuses the discussion on interval estimation of the AR in cross-sectional studies and compares the finite-sample performance of five asymptotic interval estimators of the AR by calculating the coverage probability and the average length in a variety of situations. This paper notes that the coverage probability of the two interval estimators proposed by Leung and Kupper, including the one that combines the interval estimator on the basis of Wald's test statistic, can be substantially less than the desired confidence level when the underlying risk ratio equals 1. As long as the sample size is reasonably large (> or =100) and the probability of exposure is moderate (> or =0.20), the interval estimator suggested by Fleiss can consistently perform well with respect to the coverage probability in a variety of situations considered here. However, using this interval estimator tends to generally lose efficiency. This paper also finds that with respect to the coverage probability, the interval estimator using Fieller's theorem is generally preferable to the interval estimator on the basis of Wald's test statistic when the prevalence rate ratio (RR) between the exposure and the non-exposure groups is > or =2. Finally, this paper notes that if we know that the underlying parameter RR is large (> or =4) and the probability of exposure is not small (> or =0.05), the interval estimator suggested by Leung and Kupper will probably be preferable to all the other estimators considered here. PMID- 11406843 TI - A closer look at combining data among a small number of binomial experiments. AB - In a regulatory environment, the regulators and the regulated may not be able to agree on the use of subjective prior information for a clinical trial. The use of a data-based prior offers a greater possibility for agreement, however, the degree of importance given to the prior data may still be contentious. The use of a hierarchical model to link the prior data and the current trial is shown to provide a relatively objective method for assigning weight to the prior data. Using a series of examples combining two binomial experiments, the effect of a hierarchical model on estimating rates, on the degree to which data is combined and on hypothesis testing is illustrated. In addition, the phenomenon in which combining data reduces the precision is explained. Simpler models based on finite mixtures of beta distributions are shown to work as well as the more computationally intensive, continuous mixtures. Lastly, an example combining three concurrent studies is illustrated. Published in 2001 by John Wiley & Sons, Ltd. PMID- 11406844 TI - Modelling bivariate ordinal responses smoothly with examples from ophthalmology and genetics. AB - A non-parametric implementation of the bivariate Dale model (BDM) is presented as an extension of the generalized additive model (GAM) of Hastie and Tibshirani. The original BDM is an example of a bivariate generalized linear model. In this paper smoothing is introduced on the marginal as well as on the association level. Our non-parametric procedure can be used as a diagnostic tool for identifying parametric transformations of the covariates in the linear BDM, hence it also provides a kind of goodness-of-fit test for a bivariate generalized linear model. Cubic smoothing spline functions for the covariates are estimated by maximizing a penalized version of the log-likelihood. The method is applied to two studies. The first study is the classical Wisconsin Epidemiologic Study of Diabetic Retinopathy. The second study is a twin study, where the association between the elements of twin pairs is of primary interest. The results show that smoothing on the association level can give a significant improvement to the model fit. PMID- 11406845 TI - Covariate adjustment for non-parametric tests for censored survival data. AB - We develop a method for covariate adjustment for a general class of non parametric tests for censored survival data which includes the widely used logrank and Wilcoxon tests. The covariate-adjusted tests improve the power of the unadjusted counterparts and have advantages over the covariate-adjusted Cox score test when there are outliers in the covariable space. We investigate the small sample properties of such test statistics through Monte Carlo simulations. Examples are given to illustrate the proposed procedures. PMID- 11406846 TI - Proportional hazards model for interval-censored failure times and time-dependent covariates: application to hazard of HIV infection of injecting drug users in prison. AB - Interval-censored survival data are data in which the failure times are not known precisely, but are known to lie within an interval. Such data can be analysed using a proportional hazards model with piecewise-exponential baseline hazard, a model which can be fitted by an EM algorithm easily programmed in standard statistical software. In this paper we extend the model to allow for time dependent covariates and left-truncation, and demonstrate its use by assessing the effect of imprisonment on hazard of HIV infection in a cohort of injecting drug users from Edinburgh. No conclusive effect of incarceration on hazard of HIV infection was found, but there was a suggestion that imprisonment might have been a significant relative risk factor for infection in the later period, when risk behaviour among drug users in the community was reduced. PMID- 11406847 TI - Multi-state models and outcome prediction in bone marrow transplantation. AB - Multi-state models have proved versatile and useful in the statistical analysis of the complicated course of events after bone marrow transplantation. Working from data from the International Bone Marrow Transplant Registry, we show that summary probability calculations may be useful to explore hypothetical scenarios where some transition intensities are set by the researcher. A multi-state Markov process model is specified with six states: the initial state 0; acute; chronic and both acute and chronic graft-versus-host disease A, C and AC; relapse R and death in remission D. Transition rates between the states are estimated using Nelson-Aalen estimators and Cox regression models and combined to transition probability estimators using Aalen-Johansen product integration. Besides the estimated transition probabilities to D and R we explore hypothetical probabilities obtained by artificially changing certain transition intensities, with the general purposes of getting summary views of the development for actual patients 'in this world' and of exploring the intrinsic information from real patients about consequences of various changed conditions. PMID- 11406848 TI - Rice bran oil and gamma-oryzanol in the treatment of hyperlipoproteinaemias and other conditions. AB - Diet is the first (and sometimes the only) therapeutic approach to hyperlipoproteinaemias. Rice bran oil and its main components (unsaturated fatty acids, triterpene alcohols, phytosterols, tocotrienols, alpha-tocopherol) have demonstrated an ability to improve the plasma lipid pattern of rodents, rabbits, non-human primates and humans, reducing total plasma cholesterol and triglyceride concentration and increasing the high density lipoprotein cholesterol level. Other potential properties of rice bran oil and gamma-oryzanol, studied both in vitro and in animal models, include modulation of pituitary secretion, inhibition of gastric acid secretion, antioxidant action and inhibition of platelet aggregation. This paper reviews the available data on the pharmacology and toxicology of rice bran oil and its main components with particular attention to those studies relating to plasma lipid altering effects. PMID- 11406849 TI - In vitro cytotoxicity of flavonoids against MK2 and C6 tumour cells. AB - The cytotoxic effect of five flavonoids isolated from the aerial parts of the Asteraceae family of plants were studied in vitro using LLC-MK2 and C6 glial cells. Agehoustin A was shown to be cytotoxic for both cells (71% or 67% cell death). Agehoustin B and artemetin, which lack a methoxy group at C-8 (RI), were not cytotoxic for the cells (19% and 16% cell death). The three flavonoids showed considerable suppressive effects on LLC-MK2 and C6 cell growth. Lucidin dimethylether and gnaphaliin were also not cytotoxic for cells but they had no suppressive effect on cell growth. The results show a structure-cytostatic activity relationship. We suggest that the methoxy group on C-8 and on the B ring are responsible for death cell. PMID- 11406850 TI - Hepatoprotective activity of indigtone--a bioactive fraction from Indigofera tinctoria Linn. AB - A bioactive fraction, indigtone (FA), obtained by fractionation of a petroleum ether extract of the aerial parts of Indigofera tinctoria, showed significant dose related hepatoprotective activity against CCl(4) induced liver injury in rats and mice. Hexobarbitone induced 'sleeptime', zoxazolamine induced 'paralysis time', levels of transaminases, bilirubin and total protein in serum were employed as indices of liver injury. Pre and post treatment with FA significantly reversed the majority of the parameters altered by the hepatotoxin. This indicated the preventive and restorative effect of FA in the process of CCl(4) induced liver damage. The fraction possessed a high therapeutic ratio, as no mortality was observed up to a dose of 2 g/kg p.o. in mice. PMID- 11406851 TI - In vitro antileishmanial activity of three saponins isolated from ivy, alpha hederin, beta-hederin and hederacolchiside A(1), in association with pentamidine and amphotericin B. AB - The in vitro antileishmanial activity of three saponins isolated from ivy, alpha hederin, beta-hederin and hederacolchiside A(1), was investigated on parasites of the species Leishmania mexicana, in their promastigote and amastigote forms compared with their toxicity versus human monocytes. The results showed that saponins exhibited a strong antiproliferative activity on all stages of development of the parasite but demonstrated a strong toxicity versus human cells. Association of subtoxic concentrations of saponins with antileishmanial drugs such as pentamidine and amphotericin B demonstrated that saponins could enhance the efficiency of conventional drugs on both the promastigote and the amastigote stages of development of the parasite. The results demonstrated moreover that the action of saponins on promastigote membrane was cumulative with those of amphotericin B. PMID- 11406852 TI - Effects of Dai-kenchu-to, a herbal medicine, on uterine and intestinal motility. AB - The effects of both Dai-kenchu-to and PGF(2alpha) on intestinal and uterine motility were studied in anaesthetized rabbits with force transducers implanted in the jejunum, ileum and uterus. A single intraduodenal administration of Dai kenchu-to (300 mg/kg) enhanced the intestinal motility but not the uterine motility. However, intravenous administration of PGF(2alpha) (20 microg/kg) enhanced both intestinal and uterine motility. The effects of Dai-kenchu-to on the spontaneous contraction and contractile response of the isolated rat uterine strips to oxytocin, PGF(2alpha) or ACh were also studied. Oral administration of Dai-kenchu-to at 300 mg/kg for one week had no effect on either the spontaneous contraction or the contractile response of the uterus. These results indicate that Dai-kenchu-to may exert stimulatory effects on intestinal motility, as PGF(2alpha), but has no effect on the uterine motility, suggesting a selective effect on the gastrointestinal tract. Hence, Dai-kenchu-to may be safer than PGF(2alpha) in the treatment of postoperative adhesive ileus in women. However, more studies are needed to determine whether Dai-kenchu-to could be administered to pregnant women. PMID- 11406853 TI - Long term effect of aflatoxin B(1) on lipid peroxidation in rat liver and kidney: effect of picroliv and silymarin. AB - Aflatoxin B(1) (AFB(1)) is a potent hepatotoxic and hepatocarcinogenic mycotoxin. The mechanism of cellular damage caused by AFB(1) has not been fully elucidated. Lipid peroxidation is one of the main manifestations of oxidative damage and has been found to play an important role in the toxicity and carcinogenesis of many carcinogens. The present investigation aims at assessing the comparative antioxidant effect of picroliv, a standardized iridoid glycoside fraction of Picrorhiza kurroa and silymarin, a well known standard hepatoprotective, on aflatoxin B(1) induced lipid-peroxidation in rat liver and kidney. Marked increases in lipid peroxide levels and a concomitant decrease in enzymic antioxidant levels were observed in aflatoxin B(1) (2 mg/kg, i.p) -toxicated rats, while drug (picroliv and silymarin both) treatment reversed the condition to near normal levels. The effects of picroliv and silymarin were comparable. PMID- 11406854 TI - Adaptogenic activity of a novel, withanolide-free aqueous fraction from the roots of Withania somnifera Dun. AB - The practitioners of the traditional Indian system of medicine regard Withania somnifera Dun. as the 'Indian ginseng'. A new withanolide-free aqueous fraction was isolated from the roots of this plant and was evaluated for putative antistress activity against a battery of tests such as hypoxia time, antifatigue effect, swimming performance time, swimming induced gastric ulceration and hypothermia, immobilization induced gastric ulceration, autoanalgesia and biochemical changes in the adrenal glands. This bioactive fraction exhibited significant antistress activity in a dose-related manner in all the parameters studied. The extract of Withania somnifera root (a commercial preparation available locally) was used to compare the results. A preliminary acute toxicity study in mice showed a good margin of safety. PMID- 11406855 TI - Studies on the antidiarrhoeal effect of dragon's blood from Croton urucurana. AB - The red sap obtained by slashing the bark of Croton urucurana Baill. (Euphorbiaceae), also known as dragon's blood, was screened for a possible antidiarrhoeal activity on castor oil-induced diarrhoea in rats, cholera toxin induced intestinal secretion in mice and on small intestinal transit in mice. Dragon's blood at an oral dose of 600 mg/kg caused in marked inhibition of the diarrhoeal response following castor oil administration as well as the intestinal fluid accumulation promoted by cholera toxin. At a similar dose the red sap significantly inhibited the small intestinal transit which was, however, found to be independent of the opioid mechanism. These results suggest a potential usefulness of the red sap from Croton urucurana Baill. in the control of secretory diarrhoea associated pathologies. PMID- 11406856 TI - Investigations on antimycobacterial activity of some Ethiopian medicinal plants. AB - Fifteen crude extracts prepared from seven Ethiopian medicinal plants used to treat various infectious diseases were assessed for their ability to inhibit the growth of Mycobacterium tuberculosis. A preliminary screening of the crude extracts against M. tuberculosis typus humanus (ATCC 27294) was done by dilution assay using Lowenstein-Jensen medium. None of the tested extracts except the acetone fraction obtained from the stem bark of Combretum molle (R. Br. ex G. Don.) Engl & Diels (Combretaceae) showed significant inhibitory action against this strain. The acetone fraction of the stem bark of C. molle caused complete inhibition at concentrations higher than 1 mg/mL. Phytochemical analysis of the bioactive fraction led to the isolation of a major tannin and two oleanane-type pentacyclic triterpene glycosides. The tannin was identified as the ellagitannin, punicalagin, whilst the saponins were characterized as arjunglucoside (also called 4-epi-sericoside) and sericoside. All the pure compounds were further tested against the ATCC strain. Punicalagin was found to inhibit totally growth of the ATCC and also of a patient strain, which was fully sensitive to the standard antituberculosis drugs, at concentrations higher than 600 microg/mL and 1.2 mg/mL, respectively. On the other hand, the saponins failed to show any action on the ATCC strain. It appears that our findings are the first report of tannins exhibiting antimycobacterial activity. PMID- 11406857 TI - Antiprotozoal activity of Neurolaena lobata. AB - Extracts, fractions and sesquiterpene lactones from Neurolaena lobata (L.) R. Br. (Asteraceae), a traditional medicinal plant from Guatemala, were tested in vitro against Leishmania spp. promastigotes, Trypanosoma cruzi trypomastigotes and epimastigotes and Trichomonas vaginalis trophozoites. The ethanol extract inhibited the parasite growth of L. mexicana, T. cruzi and T. vaginalis significantly. The pure germacranolides 1 and a mixture of 2 and 3, isolated from the ethonal extract, were highly active against L. mexicana and T. cruzi. PMID- 11406858 TI - The first clinical trial of topical application of procyanidin B-2 to investigate its potential as a hair growing agent. AB - Procyanidin B-2 is a compound we have identified in apple which acts as a growth promoting factor on murine hair epithelial cells. This report describes our investigation of the hair-growing effects of 1% procyanidin B-2 tonic after sequential use for 4 months. A double-blind clinical trial was performed, involving a total of 29 subjects (procyanidin B-2, 19 men; placebo, 10 men). No adverse side effects were observed in either group. In the procyanidin B-2 group, 78.9% showed an increased mean value of hair diameter, whereas only 30.0% in the placebo group showed any increase (p < 0.02, Fisher's exact probability test). The increased ratio of hairs measuring more than 40 microm in diameter after 4 months of procyanidin B-2 treatment was significantly higher than that of the placebo controls (p < 0.05, two-sample-t-test). The increase in number of total hairs in the designated scalp area (0.25 cm(2)) of procyanidin B-2 subjects after a 4 month trial was significantly greater than that of the placebo controls (procyanidin-B-2, 3.67 +/- 4.09 (mean +/- SD)/0.25 cm(2); placebo, -2.54 +/- 4.00/0.25 cm(2); p < 0.001, two-sample t-test). Procyanidin B-2 therapy shows potential as a promising cure for male pattern baldness. PMID- 11406859 TI - Biological activity of kiwifruit peel extracts. AB - Various bioactive substances in kiwifruit extracts were fractionated by organic solvent extractions, followed by silica gel and ODS chromatographies. Both cytotoxic activity and multi-drug resistance reversal activity were found in the less polar fractions. Cytotoxic activity was not always parallel the radical intensity. Antibacterial activity was distributed into various fractions and all fractions were inactive against Candida albicans and H. pylori. Only 70% methanol extracts showed anti-human immunodeficiency virus activity, and produced a broad ESR signal under alkaline conditions, in a fashion similar to lignin. These fractions also effectively scavenged O(2)(-) produced by the xanthine-xanthine oxidase reaction, suggesting a bimodal (pro-oxidant and antioxidant) action. These data suggest a medicinal efficacy of kiwifruit peel extracts. PMID- 11406860 TI - Efficacy and tolerability of a standardized willow bark extract in patients with osteoarthritis: randomized placebo-controlled, double blind clinical trial. AB - This study assessed the clinical efficacy of a chemically standardized willow bark extract in the treatment of osteoarthritis. Willow bark extract, in a dose corresponding to 240 mg salicin/day, was compared with placebo in a 2-week, double-blind, randomized controlled trial. The primary outcome measure was the pain dimension of the WOMAC Osteoarthritis Index. Secondary outcome measures included the stiffness and physical function dimensions of the WOMAC, daily visual analogue scales (VAS) on pain and physical function, and final overall assessments by both patients and investigators. A total of 78 patients (39 willow bark extract, 39 placebo) participated in the trial. A statistically significant difference between the active treatment and the placebo group was observed in the WOMAC pain dimension (d = 6.5 mm, 95% C.I. = 0.2-12.7 mm, p = 0.047); the WOMAC pain score was reduced by 14% from the baseline level after 2 weeks of active treatment, compared with an increase of 2% in the placebo group. The patient diary VAS confirmed this result, and likewise the final overall assessments showed superiority of the willow bark extract over the placebo (patients' assessment, p = 0.0002; investigators' assessment, p = 0.0073). It is concluded that the willow bark extract showed a moderate analgesic effect in osteoarthritis and appeared to be well tolerated. PMID- 11406861 TI - In vitro studies on antilithiatic activity of seeds of Dolichos biflorus Linn. and rhizomes of Bergenia ligulata Wall. AB - Soxhlet extracts of seeds of Dolichos biflorus and rhizomes of Bergenia ligulata were tested for their in vitro antilithiatic/anticalcification activity by the homogeneous precipitation method. The extracts were compared with an aqueous extract of cystone (a marketed preparation) for their activities. Also a combination of the extracts of the two plants was tested. Extracts of Dolichus biflorus showed activity almost equivalent to cystone while Bergenia ligulata showed less activity and the combination was not as active as the individual extracts. PMID- 11406862 TI - Antimicrobial study of bark from five tree species. AB - The antimicrobial activities of chloroform, methanol and aqueous extracts of the bark of Gymnanthes lucida, Gliricidia sepium, Lysiloma divaricata, Lysiloma tergemina and Coccolaba cozumelensis were tested against S. lutea, E. coli, S. epidermidis, L. monocytogenes, S. choleraesuis, S. aureus, P. aeruginosa, B. pumillus, S. typhimurium, P. vulgaris, V. cholerae and C. albicans. It was found that methanol extracts of the two Lysiloma species and G. sepium had antimicrobial effects against S. epidermidis, S. aureus, P. aeruginosa, B. pumillus and V. cholerae at doses of 200 microg. The major inhibitory effect was observed with L. tergemina which showed a bacteriostatic effect on S. epidermidis at doses of 400 microg/mL. PMID- 11406863 TI - Mutagenicity and antioxidant assessment of Stachitarpheta jamaicensis (L.) Vahl. AB - Stachitarpheta jamaicensis (L.) Vahl. is a member of the Verbenaceae commonly used in Cuba, mainly as vermifugue and against diarrhoea. The mutagenic potential of a hydroalcohol extract of its aerial parts was assessed in vitro using the Salmonella/microsome assay and in vivo in the mouse bone marrow micronucleus test. No positive response was observed in a battery of four Salmonella typhimurium strains employed: TA 1535, TA 1537, TA 98 and TA 100, when exposed to concentrations up to 5 mg/plate, with and without mammalian metabolic activation. In the same way, no increase in the micronucleus frequency in mitotic erythropoietic tissue was observed when animals were administered the extract orally in doses of 500, 1000 and 2000 mg/kg. The extract inhibited lipid peroxidation in the rat liver microsomal fraction (IC(50) = 3.6 microg/mL) but it does not seem to be an effective.OH radical scavenger (IC(50) = 76.7 microg/mL). Noteworthy, it increased in a dose dependent way the level of revertant colonies in E. coli IC 203, a strain sensitive to oxidative mutagenesis, when assayed together with hydrogen peroxide and ferrous sulphate, which suggests a pro oxidant action. PMID- 11406864 TI - Chemoprotective effect of Ixora coccinea L. flowers on cisplatin induced toxicity in mice. AB - The active fraction from Ixora coccinea flowers prevented a decrease in body weight, haemoglobin levels and leucocyte counts of mice treated with cisplatin. It also significantly prolonged the life span of cisplatin treated mice and maintained their blood urea nitrogen levels in the near normal range, indicating its chemoprotective effects. PMID- 11406865 TI - Experience with St John's Wort (Hypericum perforatum) in children under 12 years with symptoms of depression and psychovegetative disturbances. AB - The value of an extract of Hypericum perforatum (St John's wort) for children with mild to moderate depressive symptoms was investigated for the first time in a multi-centre post-marketing surveillance study. One hundred and one children under 12 years were treated for a minimum of 4 weeks with an extension to 6 weeks with parental consent and medical practitioner recommendation. the dosage used ranged from 300 to 1800 mg per day. Compliance, tolerability and efficacy were assessed every 2 weeks by physicians and parents. Based on the data available for analysis, the number of physicians rating effectiveness as 'good' or 'excellent' was 72% after 2 weeks, 97% after 4 weeks and 100% after 6 weeks. The ratings by parents were very similar. There was, however, an increasing amount of missing data at each assessment point with the final evaluation including only 76% of the initial sample. Tolerability was good and no adverse events were reported. The results of this study suggest that Hypericum is a potentially safe and effective treatment for children with symptoms of depression. PMID- 11406867 TI - Selected bibliography. PMID- 11406868 TI - [Bioethics, a path for many responses]. PMID- 11406869 TI - Mechanism of ribosomal peptide bond formation. PMID- 11406870 TI - Object processing in the infant brain. PMID- 11406871 TI - Medicare hospice benefit on the Senate's legislative agenda. PMID- 11406872 TI - Hospice care of the future ... now. PMID- 11406873 TI - The many faces of pain for older, dying adults. AB - An integration of knowledge from the fields of geriatrics, pain management, and palliative care is needed to ensure adequate pain control for the older adult who is dying. An overview is provided of the multiple factors (i.e., chronic illness, malignant disease, care procedures, emotional and cognitive status, response of others) that can cause and exacerbate pain at the end of life for the elderly. Treatment considerations are discussed and an illustrative case study is presented. PMID- 11406874 TI - The business of palliative medicine: management metrics for an acute-care inpatient unit. AB - For any palliative medicine inpatient unit to be economically viable, certain management metrics need to be followed. Palliative medicine can provide both a compassionate and economical service within the current acute inpatient hospital environment. In this article, we will review the administrative and financial factors we have identified that influence the business of acute palliative medicine. PMID- 11406875 TI - A critical pathway for interdisciplinary hospice care. AB - In today's managed health care environment, many hospices are seeking to streamline care while maintaining quality. Quality hospice care is dependent on communication and planning among members of the interdisciplinary team. A critical pathway can be useful in determining effective interventions in a uniform manner. In order to maintain quality and provide uniform care, a guiding framework is necessary. A critical pathway is presented here and is designed specifically for hospice care. An overview is presented of the usefulness of the critical pathway in the case management model. In theory, the critical pathway can be used as an outline for care to improve quality, maximize effectiveness, and generate further research in hospice practice. PMID- 11406876 TI - Dealing with AIDS-related loss and grief in a time of treatment advances. AB - Recent advances in the treatment of HIV/AIDS have dramatically changed the lives of many patients and their loved ones as well as those who care for them. However, not all patients respond successfully to the latest treatments. Hospice professionals need to understand the experience of AIDS patients and their loved ones in this period of hope and disappointment, and to find appropriate ways to support and care for them. This article explores the implications of treatment advances for AIDS patients, their loved ones, and professionals in dealing with loss and grief. PMID- 11406877 TI - Mirtazepine: heir apparent to amitriptyline? AB - "What's new in therapeutics?" will examine and evaluate drugs that may have a place in hospice, palliative, and long-term care. Mirtazepine will be examined and evaluated. Mirtazepine is a potential alternative anti-depressant with multiple additional benefits. It is an atypical anti-depressant, which has both noradrenergic and specific serotonergic receptor antagonism (NaSSa), and a unique pharmacological profile. Mirtazepine appears to be a "designer drug" for palliative medicine with a number of benefits, but cost may be a drawback. PMID- 11406878 TI - Transdermal fentanyl in the hospice: a survey of rescue dosing and pain control. AB - The case records of 25 patients who received transdermal fentanyl as a primary analgesic during routine hospice care were surveyed for pain control and rescue medication use. The majority of patients (76 percent) had cancer-related pain and were treated in hospice for an average of approximately 30 days. Most received oral medications for supplemental rescue analgesia. During the sampling periods, on average, pain intensity was reported as mild. Over the same periods, patients required a mean of 6.1 (+/- 0.7) doses of rescue medication per day, with a range of zero to 12 doses per day. Five patients required rescue dosing every two hours on some treatment days. Although adequate pain control was generally accomplished with transdermal fentanyl in the group as a whole, the frequency of rescue dosing outside of the initial titration period appears unacceptably high. PMID- 11406879 TI - Pain and aging. PMID- 11406880 TI - Methadone-induced myoclonus in advanced cancer. AB - Methadone is recommended as being free of some of the neuropsychological side effects noticed with morphine, which are attributed to active metabolites. A patient that received methadone for cancer-associated pain developed myoclonus as a side effect. This has rarely been reported before in the literature. The pathophysiology and management of myoclonus are discussed. PMID- 11406881 TI - Caring across "cultures". PMID- 11406882 TI - Euthanasia: can the trend be countered? PMID- 11406883 TI - Switching to methadone. PMID- 11406884 TI - Existential suffering and palliative sedation: a brief commentary with a proposal for clinical guidelines. PMID- 11406885 TI - OxyContin, the media, and law enforcement. PMID- 11406886 TI - Medicare hospice per diem rate increased from $90 to $95. PMID- 11406887 TI - The high cost of at-home caregiving. PMID- 11406888 TI - Oregon's use of Death With Dignity Act remains steady in its third year. PMID- 11406889 TI - Mt. Sinai physicians suggest strategies and solutions for palliative care. PMID- 11406890 TI - Conversion ratio and cost of oxycodone. PMID- 11406891 TI - Additional variables in the appropriateness of fentanyl. PMID- 11406892 TI - Alleviating the suffering of seriously ill children. AB - Modern medicine has largely focused on the physical aspects of disease, aggressively attacking the illness, often at the expense of caring for pain and suffering. Medical interventions based solely on the diagnosis and treatment of disease limit the medical care of the severely ill child. Such an approach is particularly detrimental when caring for the terminally ill. Successful care of children with chronic, life-threatening, or terminal illnesses requires a comprehensive assessment of their physical, psychological, and spiritual needs as well as a process of collaboration between members of the multiple disciplines involved in the care of the patient and the family unit as a whole. Supportive/palliative care serves as a bridge between a scientific (disease oriented) and humanistic (person-oriented) approach to patient care. Bridging this gap early in the course of life-threatening illness is essential for successful palliative intervention to relieve suffering and improve the quality of life for the child and his or her family. A model that introduces supportive, palliative, and hospice services into the mainstream of medical therapy is emphasized as a standard for the care of all children with significant chronic, life-threatening, or terminal illness. This article expands on a previous paper published in the American Journal of Hospice & Palliative Care (Kane JR, Barber RG, Jordan M, et al.: Supportive/palliative care of children suffering from life threatening and terminal illness. May/June 2000; 17(3): 165-172). PMID- 11406893 TI - Measuring comfort in caregivers and patients during late end-of-life care. AB - The purpose of this study was to test several formats of end-of-life comfort instruments for patients and closely involved caregivers. Kolcaba's Comfort Theory was the theoretical framework utilized. Different response formats for two end-of-life (EOL) comfort questionnaires (for patients and caregivers, respectively), and horizontal and vertical visual analog scales for total comfort (TC) lines were compared in two phases. Evaluable data were collected from both members of 38 patient-caregiver dyads in each phase. Suitable dyads were recruited from two hospice agencies in northeastern Ohio. Cronbach's alpha for the EOL comfort questionnaire (six response Likert-type format) tested during phase I for patients was .98 and for caregivers was .97. Test-retest reliability for the vertical TC line tested during phase I for patients was .64 and for caregivers was .79. The implications of this study for nursing practice and research are derived from the American Nursing Association (ANA) position statement about EOL care, which states that comfort is the goal of nursing for this population. These instruments will be useful for assessing comfort in actively dying patients and comfort of their caregivers as well as for developing evidence-based practice for this population. PMID- 11406894 TI - Pharmacy student training in United States hospices. AB - Hospice is a quickly growing field in health care in the United States. As the pharmacist's role in providing patient care to persons at the end of life increases, considerations should be given for training pharmacy students in this area. The objectives of this study were to examine the frequency of pharmacy student education and training among United States hospice organizations as well as to describe factors of hospice organizations that are associated with pharmacy student training. This is the first study of which we are aware to address the availability of experiential rotations for pharmacy students in hospice programs. A one-page questionnaire was mailed to 3,762 hospice organizations with addresses obtained from the National Hospice and Palliative Care Organization (NHPCO). Following two mailings, eight weeks apart, 907 responses were obtained. Ninety four (10 percent) hospices trained pharmacy students, 246 (27 percent) trained medical students, 357 (39 percent) trained social work students, and 623 (69 percent) trained nursing students. These results indicate that the experiential training needs of United States pharmacy students are being addressed. However, further study is warranted to describe the various experiences of pharmacy students within the hospice setting. PMID- 11406895 TI - Methylphenidate for fatigue in advanced cancer: a prospective open-label pilot study. AB - Psychostimulants such as methylphenidate are used for fatigue in cancer patients. We report a prospective, open-label, pilot study of the successful use of methylphenidate to treat fatigue in nine of 11 consecutive patients with advanced cancer. Seven had received radiation or chemotherapy, a median of three weeks (range from one to 30 weeks) prior to methylphenidate. A rapid onset of benefit was noted, even in the presence of mild anemia. Sedation and pain also improved in some. Only one patient had side effects severe enough to stop the medication. PMID- 11406896 TI - Performing a medication regimen review in hospice and palliative care. PMID- 11406897 TI - A German model for methadone conversion. PMID- 11406898 TI - The terminally ill: dying for palliative medicine? PMID- 11406899 TI - The role physicians play throughout the birthing process. AB - The second of a series, this study examines the effect of physician service performance on women's perceptions of physician quality before, during, and after delivery. We also examine changing physician-patient relationships throughout the birthing experience and compare our findings to those reported in a previous study that focused on nurse-patient relationships. Our findings show that physicians can have a profound impact on patient quality assessments and that these perceptions are prone to change as patients progress through the three birthing stages. PMID- 11406900 TI - 25 marketing assumptions for new economy health care leaders. PMID- 11406901 TI - What the DTC movement means to health care. AB - The direct-to-consumer (DTC) movement is gaining momentum as the health care industry becomes more consumer driven and the ability to target narrow population segments is better refined. To fully benefit from DTC marketing, health care marketers must modify their methodologies and recognize the promise represented by 280 million potential customers. This radical approach requires throwing out some preconceived ideas about marketing and learning to treat health care consumers as individuals with unique lifestyle traits. PMID- 11406902 TI - Breaking through language barriers. PMID- 11406903 TI - Developing a profile of LASIK surgery customers. PMID- 11406904 TI - Hospitals and the Web: a maturing relationship. PMID- 11406905 TI - Taking the pulse of Internet pharmacies. AB - Like most businesses, online pharmacy companies will only be successful if they make sure customers are satisfied with the service they receive. But what attributes of service quality lead to satisfaction and dissatisfaction? This study identified 19 Internet pharmacy service quality dimensions in three categories: (1) product cost and availability, (2) customer service, and (3) the online information system. Our analysis uncovered attributes that tend to determine consumer satisfaction and points out ways to improve overall service quality in the Internet pharmacy arena. PMID- 11406906 TI - Angina in the elderly: the need for a balanced approach. PMID- 11406907 TI - Antihypertensive efficacy and tolerability of low-dose perindopril/indapamide combination compared with losartan in the treatment of essential hypertension. AB - The aim of the study was to evaluate the antihypertensive efficacy and tolerability of the low-dose combination of the angiotensin-converting enzyme inhibitor perindopril 2 mg plus the diuretic indapamide 0.625 mg (P/I) compared with the angiotensin II antagonist losartan 50 mg (L50) in the treatment of essential hypertension. Patients (n = 277) were randomised, double-blind and allocated to receive either P/I or L50 once daily for a period of 12 weeks. Responder and normalisation rates in the two groups were compared by a chi 2 test. Ambulatory blood pressure monitoring results were compared using the one tailed Student's t-test. Normalisation rates were significantly greater in the P/I group (76.0%) than in the L50 group (60.0%) (p = 0.009). Responder rates were significantly higher in the P/I group (91.7%) than in the L50 group (81.8%) (p = 0.025). The average blood pressure reductions were: in sSBP (P/I-L50 = -2.4 mmHg; CI: 6.2; 1.3) and sDBP (P/I-L50 = -2.0 mmHg; CI: -4.2; 0.2). The average night time SBP decrease (ABPM) was significantly greater in the P/I group (p = 0.041). The tolerability was comparable between the two groups in terms of emergent adverse events related to treatment (12.4% for P/I patients and 8.4% for L50 patients). Laboratory evaluations did not show any significant variations. It was concluded the low-dose P/I combination had significantly higher responder and normalisation rates than L50. This study also confirmed the good tolerability of both treatments. PMID- 11406908 TI - DICE 2: a further investigation of the effects of chance in life, death and subgroup analyses. AB - In an investigation into how chance might influence the distribution of deaths in a randomised trial and the time of those deaths, and to highlight the possible dangers of subgroup analyses, 100 randomised controlled trials were simulated and 50 subgroup pairs were simulated for some of these trials. Each of 580 control patients from a colorectal cancer trial was randomly coded to simulate allocation to treatment or control, the main outcome measure being time to death. Not surprisingly, most of the 100 trials gave non-significant results. Four were conventionally significant with logrank 2p-values of less than 0.05. The most extreme result was associated with a logrank 2p-value of 0.003, showing an absolute reduction in four-year mortality of 40% (SD 15) for patients allocated to treatment. One of the simulated prognostic factors for this trial (subgroup 13) showed that mortality for one type of patient was non-significantly slightly increased by treatment, whereas treatment reduced four-year mortality by 64% (SD 16) among the other patients in the trial (2p = 0.00006). Similar, extreme results were found for a trial of borderline statistical significance overall. Chance can influence the overall results of any randomised controlled trial, regardless of how well it is conducted, and can play an even more powerful role in the results of subgroup analyses. This should be borne in mind both by trialists when reporting their results and by readers and reviewers of those reports. PMID- 11406909 TI - A cost-effectiveness model of alternative statins to achieve target LDL cholesterol levels. AB - An economic model was developed to estimate the relative cost-effectiveness of alternative HMG-CoA reductase inhibitors (statins)--atorvastatin, cerivastatin, fluvastatin, pravastatin and simvastatin--to achieve target low-density lipoprotein cholesterol (LDL-C) levels in a population of secondary CHD prevention patients. By using a cholesterol target as the endpoint of interest and a dose titration approach, the model assumes that the statins demonstrate a class effect through cholesterol lowering. The model was used to estimate the proportion of patients achieving target LDL-C levels (< 3 mmol/l) under each scenario tested. Total costs and incremental cost-effectiveness relative to no treatment and to the lowest cost option were estimated for each scenario. Total costs were highest for pravastatin and lowest for cerivastatin. Compared with no treatment, the incremental cost per patient treated to target LDL-C varied between 383 Pounds (atorvastatin) and 1213 Pounds (pravastatin). Incremental cost effectiveness ratios in comparison with the lowest cost treatment (cerivastatin) were 141 Pounds per additional patient achieving target LDL-C with atorvastatin, and 275 Pounds with simvastatin. Fluvastatin and pravastatin were both less effective and more expensive than the lowest cost therapy. Although cerivastatin was associated with lowest expected costs, therapy with atorvastatin achieved the lowest cost-effectiveness ratios. Hence atorvastatin would allow the largest number of patients to be treated to target LDL-C within a fixed drug budget. Choosing between drug therapies on the basis of price alone may be misleading if the effectiveness of therapies varies. PMID- 11406910 TI - Comparison of clinical characteristics and high-risk factors in Australian aboriginal and non-aboriginal neonates with necrotising enterocolitis. AB - In a five-year retrospective data analysis, incidence of > stage II necrotising enterocolitis (NEC) was four times higher in aboriginal (18/125) than non aboriginal (11/306) neonates, all < or = 32 weeks' gestation. Stage III NEC occurred more frequently (10/18 vs 3/11) and related mortality was higher (44.4% vs 0%) in aboriginal than non-aboriginal neonates. Risk factors for NEC-prolonged rupture of membranes (50% vs 9.1%, p = 0.01), prematurity (median [range] gestational age: 25 (24-28.5) vs 30 [27-33]) weeks, p = 0.02), birth weight (< 1 kg 13 [72%] vs 2 [18%], p = 0.007) and intrauterine growth retardation (50% vs 0%, p = 0.01) occurred more frequently in aboriginal neonates. Though feed type and increments per day were similar, aboriginal neonates received higher volume of feeds/kg/day (median [range]: 154 [145-189] vs 106 [103-134] ml, p < 0.05). Condition at delivery and respiratory status before development of NEC were not significantly different. PMID- 11406911 TI - Combination therapy in angina: a review of combined haemodynamic treatment and the role for combined haemodynamic and cardiac metabolic agents. AB - The 1997 European Society of Cardiology guidelines emphasised medical therapy as the mainstay of treatment of stable angina. Until recently, the antianginal drugs available to control symptoms (nitrates, beta-blockers and calcium antagonists) have all been haemodynamic agents that eliminate or reduce angina attacks principally by decreasing myocardial oxygen demand. They may also improve myocardial perfusion. The haemodynamic agents are often used in combination. However, there is conflicting evidence for the efficacy of combination treatment. Two recently published large clinical studies, the Total Ischaemic Burden European Trial (TIBET) study and the International Multicenter Angina Exercise (IMAGE) study, suggest that combined haemodynamic treatment is no more effective than optimal monotherapy. The results from IMAGE suggest that most of the additional effect observed with combined therapy was attributable to recruitment by the second drug of patients who had not responded to monotherapy, and that most patients continued to have a disturbing frequency of anginal attacks after receiving combined therapy. Metabolic agents are a new class of drugs that directly modify the use of energy substrates in the heart, lessening ischaemic injury and improving cardiac performance during ischaemia. Due to their non haemodynamic mode of action, metabolic agents such as trimetazidine or L carnitine may provide independent benefit in ischaemia when used as monotherapy or additional benefit when used in combination with a conventional agent. Clinical trials have shown that combined haemodynamic and metabolic treatment is more effective than combined haemodynamic therapy and is well tolerated. It is suggested that combination haemodynamic and metabolic therapy is a logical new approach to patients whose angina is inadequately controlled despite optimal haemodynamic therapy. PMID- 11406912 TI - Aetiology and treatment of hyperhomocysteinaemia causing ischaemic stroke. AB - Recent studies have shown that hyperhomocysteinaemia is a common, independent and easily modifiable risk factor for atherosclerotic and thromboembolic diseases such as cerebrovascular disease, coronary artery disease and venous thrombosis. The vascular risk rises continuously across the spectrum of elevated plasma homocysteine concentrations. It is at least as important as cholesterol, lipoprotein abnormalities and hypertension and should be part of risk assessment, especially those at high risk. Moderately elevated plasma homocysteine concentration is readily correctable by folic acid, betaine, or vitamin B12 supplementation. It seems logical to assume that a reduction in homocysteine concentration will reduce the risk of ischaemic stroke, but there are as yet no published data to prove this. This review will discuss the aetiology and possible treatment of hyperhomocysteinaemia causing ischaemic stroke. PMID- 11406913 TI - C-reactive protein. AB - C-reactive protein (CRP) is a protein whose concentration in serum is increased in response to inflammatory stimuli. Increased levels serve to identify organic disease, monitor disease activity and assist differential diagnosis. High values are observed early in bacterial infections, active rheumatoid disease, Crohn's disease, acute myocardial infarction and after major trauma. In patients with ischaemic chest pain, a raised CRP value on hospital admission is associated with an adverse prognosis. In apparently healthy individuals, a raised CRP value indicates an increased risk of developing atherosclerotic vascular disease, but also increased benefit from aspirin prophylaxis and treatment of hyperlipidaemia. PMID- 11406914 TI - A study of the appropriateness of antibiotic use in the medical wards of a tertiary teaching hospital in Malaysia. AB - A prospective study was conducted to determine the proportion of patients who received an antibiotic within 12 hours of admission to the medical wards. During the four-week study, 234 patients were admitted to medical wards from casualty; 68 patients (29%) received an antibiotic. The survey indicated that antibiotics were inappropriate in 22-65% of those treated. This study also shows that as many as 67% of patients who received intravenous antibiotics could have been equally well treated with oral preparations. PMID- 11406915 TI - Risedronate: clinical usage. AB - Risedronate is a new bisphosphonate--a family of drugs that inhibit bone resorption--and thus can be used in various bone conditions involving increased levels of bone resorption, such as postmenopausal osteoporosis, glucocorticoid induced bone loss, and Paget's disease of bone. In placebo-controlled clinical trials, risedronate has been shown to prevent bone loss in postmenopausal women, to decrease the incidence of vertebral and non vertebral (including hip) fractures in postmenopausal women with osteoporosis, and to prevent bone loss in men and women treated with moderate to high doses of glucocorticoids. Risedronate has also been shown substantially to decrease the severity of bone pain and the level of bone turnover in Paget's disease of bone, and to improve the radiographic lesions of this disease. Risedronate is safe and well tolerated. Thus, risedronate is a new option for the management of postmenopausal osteoporosis, Paget's disease of bone and corticosteroid-induced bone loss. PMID- 11406916 TI - Images in cardiology: left ventricular free wall rupture. AB - Left ventricular free wall rupture is thought to account for approximately 15% of deaths following acute myocardial infarction (MI). We present a case of left ventricular free wall rupture in a 64-year-old man following a lateral MI. The diagnosis was made with 2D and colour flow Doppler echocardiography. Unfortunately the patient died before emergency surgery could be performed. PMID- 11406917 TI - Polyserositis as a rare component of polyglandular autoimmune syndrome type II. AB - Polyglandular autoimmune (PGA) syndromes (types I and II) may affect various endocrine and non-endocrine organs in the body. In the commoner PGA type II, primary adrenal insufficiency, autoimmune thyroid disease and type I diabetes mellitus are the most frequent manifestations. Serositis with pericardial or pleural involvement is not a well known component of the disease. Here, we report a 21-year-old man who first presented with a pleuropericardial effusion and Graves' disease, and who then developed type I diabetes mellitus. PMID- 11406918 TI - An unusual presentation of thyroglossal duct cyst. AB - Thyroglossal duct cyst usually presents as a painless swelling in the mid-line of the neck. This is the first documented case of an acute presentation of hoarseness and multi-loculated swelling. It also contrasts the limitations of ultrasonography with the accuracy of magnetic resonance imaging in such atypical swellings. PMID- 11406919 TI - Synchronous colonic tumours of dual pathology. AB - Synchronous colonic tumours of dual pathology are extremely rare. A review of the literature revealed that few cases have been reported to date. Because of their rarity and lack of specific symptoms, preoperative diagnosis is not easy and there is no protocol as yet for the ideal management of these cases. We present such a case which was treated by a combination of surgery and chemotherapy. PMID- 11406920 TI - Endometrioma of the liver. AB - Hepatic endometriosis is extremely rare. The diagnosis is often missed and treatment is delayed. A 37-year-old woman was referred to the gynaecology department with vague abdominal pains. She had a past history of pelvic endometriosis and hysterectomy with bilateral salpingo-oophorectomy. Further investigations in due course confirmed it to be endometriosis of the liver. She was eventually referred to the hepatobiliary unit, almost three years after her first presentation, where she was operated on with good results. PMID- 11406921 TI - Meeting the challenges of vascular dementia. Introduction. AB - Ongoing controversy over the challenges associated with the diagnosis and management of patients with vascular dementia (VaD) or Alzheimer's disease with cerebrovascular disease (CVD) has led to difficulties with establishing epidemiology and diagnostic guidelines, as well as deciding upon optimal management practices. In March 2001, an international group of physicians and researchers gathered to resolve some of these issues and provide clinicians with useful information relating to this large patient group. In addition to simple coexistence, VaD and Alzheimer's disease appear to share common pathophysiological mechanisms, such as neurotransmitter abnormalities. Treatments that target the common pathophysiology of the two conditions may prove useful. To date, no reports have been published of any agent demonstrating broad efficacy (on cognitive and noncognitive outcomes) in VaD and Alzheimer's disease with CVD in randomised, controlled trials. Recent developments in understanding the pathophysiology of the conditions, however, have taken researchers an important step closer to finding an effective treatment, not only for patients with Alzheimer's disease, but also for those with coexisting CVD and those with VaD. PMID- 11406922 TI - Clinical deficits of Alzheimer's disease with cerebrovascular disease and probable VaD. AB - Vascular dementia (VaD) describes dementia arising from cerebrovascular disease (CVD) and ischaemic brain injury, and relates to a number of different vascular mechanisms and clinical manifestations. The characterisation of brain lesions by neuroimaging analysis, and the study of their relation to clinical deficits such as cognitive and functional decline, are critical to the concept and treatment of VaD, and form an important part of widely used diagnostic criteria for this dementia type. For instance, the extent to which pathological brain lesions cause, compound or coexist with cognitive impairment is a major determinant of other clinical deficits, their nature and the rate of disease progression. Through numerous neuroimaging and epidemiological studies, VaD is now seen to encompass a heterogeneous group of clinical syndromes such as multiple-infarct (cortical) dementia, small-vessel (subcortical) dementia and, less commonly, dementia associated with strategic infarcts. Due to a large number of similarities in clinical symptoms, pathophysiological mechanisms, associated risk factors and neurochemical deficits between VaD and Alzheimer's disease, patients with coexistent Alzheimer's disease and CVD ('mixed' dementia) represent another important, but previously underestimated subgroup. This article reviews the clinical symptoms and neuroimaging findings most commonly observed in patients with VaD. Increased familiarity with the clinical picture of VaD should offer more hope of defining realistic treatment aims for future pharmacotherapy. PMID- 11406923 TI - The pharmacological rationale for treating vascular dementia with galantamine (Reminyl). AB - There is considerable evidence indicating that, as in Alzheimer's disease, the central cholinergic system is impaired in vascular dementia (VaD). Using lessons learned from Alzheimer's disease research, it has been proposed that enhancement of the cholinergic system is a rational approach to treating the symptoms of VaD. Galantamine's dual mode of action may provide a greater chance of success in treating patients with Alzheimer's disease through enhanced efficacy on the cognitive, functional and behavioural aspects of dementia. Trials are currently underway to see if this broad spectrum of efficacy extends to patients with Alzheimer's disease with cerebrovascular disease ('mixed' dementia) or VaD, as well as other conditions, and the results are eagerly awaited. PMID- 11406924 TI - Outcome measures for probable vascular dementia and Alzheimer's disease with cerebrovascular disease. AB - Vascular dementia (VaD) can be defined as dementia associated with cerebrovascular disease (CVD), and accounts for a large proportion of all dementia cases. There is substantial overlap in the clinical symptomatology, pathophysiology and neurochemical mechanisms in VaD compared with Alzheimer's disease, suggesting that an effective treatment for Alzheimer's disease may also offer benefit as a symptomatic treatment in VaD. However, there are currently no explicit guidelines for conducting clinical pharmacotherapy trials in VaD patients. Two important requirements for assessing therapeutic benefits in such trials are 1) the inclusion of appropriate patients and 2) the use of appropriate outcome measures. Debate on the precise definition of VaD in relation to patient selection criteria continues, but many of the recommendations for outcome measures in Alzheimer's disease are already applicable to VaD. There is consensus that cognitive and global function measures, and assessments of abilities to perform activities of daily living (ADL) must be included as part of the optimal assessment battery in VaD trials. A measure of reduced behavioural symptoms with associated reductions in demands on caregivers would also be desirable. However, care must be taken in extrapolating Alzheimer's disease-specific evaluations to VaD, in that important differences in specific domains affected and characteristics of disease course must be taken into account. Between them, measures such as the Alzheimer's Disease Assessment Scale-cognitive subscale (ADAS-cog; perhaps with supplemental tests of attention and other frontal lobe functions), evaluations of clinical global impression of change and a functional assessment addressing instrumental as well as basic ADL, e.g. Disability Assessment in Dementia (DAD) scale, should provide a good overall description of VaD-related deficits and sufficient appraisal of treatment effects. The Neuropsychiatric Inventory has also been shown to have good potential utility for measuring behavioural alterations in VaD. These and other assessments are reviewed to provide a balanced and realistic view of the type of treatment outcomes that can be expected in VaD pharmacotherapy trials, and to address the best ways of measuring these outcomes. PMID- 11406925 TI - Public health importance of vascular dementia and Alzheimer's disease with cerebrovascular disease. AB - Vascular dementia (VaD) refers to a heterogeneous group of conditions that include all dementia syndromes resulting from ischaemic, haemorrhagic, anoxic or hypoxic brain damage. VaD is the second most common cause of dementia in the elderly after Alzheimer's disease. Persons with VaD are at greater risk of morbidity and mortality compared with those without dementia or those with Alzheimer's disease, and appear to be at greater risk of institutionalisation. Despite the importance of the problem posed by VaD, few placebo-controlled, double-blind, randomised clinical trials have been conducted. Although dementia may result solely from the accumulation of brain damage from cerebrovascular disease (CVD), recent data suggest that VaD often results from a combination of both CVD and Alzheimer's disease ('mixed' dementia). This raises the possibility that persons with VaD may respond to medications that are commonly used to treat Alzheimer's disease. PMID- 11406926 TI - What is vascular dementia? AB - Cerebrovascular disease (CVD) and dementia frequently coexist in elderly patients. The question of whether the CVD causes the dementia depends on how 'dementia' is defined. Traditional definitions specified that dementia involved a decline in intellectual ability as a core feature. However, revised definitions have since stipulated two key elements: 1) a global rather than focal neurobehavioural deficit and 2) impairment in activities of daily living (ADL). When applied to CVD, these latter concepts of dementia raise difficulty: Focal cerebrovascular lesions in the cortex generate location-specific neurobehavioural deficits that are part of the dementia syndrome, but, even in combination, do not represent a global intellectual decline. Most cerebrovascular lesions are associated with physical symptoms that make it difficult to evaluate whether cognitive impairments have an independent impact on ADL. The majority of neurobehavioural symptoms in CVD are caused by small-vessel-type subcortical lesions and are dissimilar to those seen in Alzheimer's disease. There are several pathogenetic mechanisms, however, by which large-vessel or small-vessel CVD can cause global cognitive and intellectual impairments, allowing a diagnosis of vascular dementia (VaD): An accumulation of ischaemic lesions in the cortex may produce global intellectual impairment, particularly if they affect important areas of the brain. Single small infarcts, or haemorrhages in strategic subcortical locations, may interfere with specific circuits connecting the prefrontal cortex to the basal ganglia, or with nonspecific thalamocortical projections. This may generate combinations of executive dysfunction, personality change or apathy, which are associated with hypoperfusion and hypometabolism predominantly in frontal cortical areas. Extensive white matter lesions probably affect cognitive function through a loss of axons, producing a functional disconnection of the cortex. This can manifest as significant reductions in blood flow and metabolism in frontal, temporal and parietal cortical areas, which do not show any structural damage. Given the diversity of aetiological factors, pathological changes and pathogenetic mechanisms associated with VaD, several distinct syndromes must be distinguished. Further study is needed to demonstrate that this emerging concept can improve diagnosis, guide treatment and stimulate research. PMID- 11406927 TI - Diagnosis of vascular dementia and Alzheimer's disease. AB - Vascular dementia (VaD) and Alzheimer's disease are sometimes difficult to distinguish due to overlaps in symptomatology, pathophysiology and comorbidity. The issue of differential diagnosis is further complicated by the fact that many patients have concomitant Alzheimer's disease and cerebrovascular disease (CVD) ('mixed' dementia). Each pathology may contribute to varying degrees, giving rise to a continuum of patients in whom pure CVD and pure Alzheimer's disease represent the two extremes. Despite the clear overlap between the conditions, and the prevalence of 'mixed' dementia, a number of criteria for Alzheimer's disease and VaD do not make provision for 'mixed' dementia distinct from the coincidence of any two other dementing illnesses. We will discuss the current diagnostic criteria for VaD, with or without coexisting Alzheimer's disease, in an effort to determine how best to diagnose VaD. These include traditional criteria such as the Diagnostic and Statistical Manual of Mental Disorders or the Hachinski Ischemic Scale, and the more recently developed criteria by the California Alzheimer's Disease Diagnostic and Treatment Centers (CAD-DTC) and the National Institute of Neurological Disorders and Stroke and the Association Internationale pour la Recherche et l'Enseignement en Neurosciences (NINDS-AIREN) International Workshop. The CAD-DTC and NINDS-AIREN rely on neuroimaging--ideally, every patient suspected of dementia should have brain imaging, but although this is possible in clinical trials (for which these criteria were designed), it is not always feasible in population-based epidemiological studies and clinical practice in some countries. PMID- 11406928 TI - [Breech presentation: the last days of the low method?]]. PMID- 11406929 TI - [Medically assisted reproduction and the desire for a child by HIV infected couples: has the time for a change in attitude come?]]. AB - Today, in developed countries, many HIV-infected people remain in good health thanks to antiviral medication. A growing number of them want to have children. Medical possibilities for preventing contamination of the partners of seropositive men through assisted reproduction and of children thanks to antiviral medicines during pregnancy are summarized. These changes result in ethical considerations which lead the authors to question the conventional systematic medical advise against pregnancy and has encouraged them to assist reproduction for a number of these couples. Today, the balance between the importance of the message of prevention and the benefit for patients of being assisted in their desire for a child has tilted towards medical intervention. It would seem legitimate today to intervene in the most favourable situations rather than see these couples take the risk of spontaneous conception outside health care structures. This implies to adapt medical structure (separate laboratory, appropriate procedure, precise protocols). This approach, which is coherent from the scientific point of view, respects both the autonomy of people carrying HIV as well as the essential interest for the child, in "being" born uninfected and also has the enormous advantage of allowing access to parenthood without destroying the consistency of the message of prevention of sexual contamination. PMID- 11406930 TI - [Ovarian fibrothecal tumors. Apropos of 12 cases]. AB - OBJECTIVE: To identify diagnosis, therapeutic and histological particularities of ovarian fibro-thecomas. STUDY DESIGN: A retrospective study of 12 patients who underwent surgical treatment for ovarian fibro-thecoma between January 1994 and December 1999 was reported. Clinical, ultRasonographic, tumour marker, therapeutic and histologic data were analysed. RESULTS: Mean age of patients was 46.83 years. Three patients were in perimenopausal period and six were menopausal. Ultrasonographic findings were: ovarian echogenic tumor in six cases, hypoechogenic tumor in three cases, mixed tumor in three cases and anechogenic tumour in one case. CA 125 level measured in 11 cases was normal. Nine patients underwent laparotomy. Three patients underwent laparoscopy, however, one was converted to laparotomy due to a suspected tumour. A conservative treatment was performed for three patients and radical treatment for nine patients. Histological findings were eight fibromas and five fibro-thecomas. CONCLUSION: Ovarian fibro-thecoma is uncommon. These lesions occur often in perimenopausal and menopausal patients. Clinical, ultrasonographic and tumour marker data remains the best preoperative approach currently available for ovarian tumours. However, the diagnosis remains histological. Tumorectomy is well indicated for young patients, however radical treatment is indicated for perimenopausal and menopausal patients. PMID- 11406931 TI - [Pelvic hydatid yst: apropos of 8 cases]. AB - The hydatidosis fisues in the endemic state in Morocco, its pelvigenital localization is rare and doubtful. The objective of this study is to examine the epidemio-clinic appearance, the diagnosis means and the therapeutic flow of the pelvigenital hydatid cyst in Morocco. This retrospective study is about eight patients hospitalized and cured for pelvigenital hydatid cyst in lalla Meryem maternity of Casablanca during a period of six years (1992-1997). The hydatid cyst diagnosis was confirmed by anatomopathology. This affection represented 0.80% of the pelvic mass operated during the same periode. The majority of our patients was from rural origin, their age varies between 22 and 70 years old. The discovery circumstances were dominated by abdomino-pelvic mass. The diagnosis was nearly certain prior to the surgery for half of the cases. The genital organs were the most reached by the pelvic hydatidosis (75%). The treatment was purely surgical, adapted to each case in according to the localization, the volume and the contribution of the cyst. The medical treatment has been prescribed in one case in post surgery. The evolution was good in all the cases. The hydatidos cyst must always be present in mind when dealing with cyst pelvic formation in an endemic country like Morocco. The prophylaxia preserves its important value and must constitute a hinder against this disease in endemic countries. PMID- 11406932 TI - [Opinions of couples on care during medical termination of pregnancy]. AB - OBJECTIVES: To study the medical care, staff attitudes and patients' satisfaction from the decision to the post-intervention medical visit for termination of pregnancy for fetal abnormalies. PATIENTS AND METHODS: All patients and their spouses having a termination of pregnancy at the "Unite de Medecine Foetale" in Port-Royal Hospital between November 1996 and July 1997 were contacted for the study. A self-administered questionnaire was mailed six to eight weeks after intervention. Forty seven women and 42 men returned a completed questionnaire, the response rates were respectively 68% and 61%. RESULTS: The patients and their spouses rated globally very high their satisfaction about the care received. The delay before intervention, the length and pain of labour were rated less positively. The factors associated with satisfaction were the quality of the relationship with the staff, and of information. Positive feelings about delivery were linked with the consideration and relief of pain. Most respondents mentioned that their physical and psychological state has improved at the moment of the survey but the psychological distress subsisted or has increased in one fourth of the cases. On the whole the answers made within the couples were correlated. CONCLUSION: The positive results should be moderated by the number of non respondents. In a context of very high rates of satisfaction, psychological distress is still present for one respondent out of four, six to eight weeks after termination of pregnancy for fetal abnormalies. PMID- 11406933 TI - [Detection of fetal cells in maternal blood: myth or reality?]. AB - Fetal cells exist in maternal blood and can be utilized for prenatal diagnosis. These cells are present from the sixth week of gestation, with frequency increasing as gestation advances, to many years after the birth. Enrichment of trophoblast cells, erythroblasts and lymphocytes was performed with various density gradient techniques and either magnetic activated or fluorescent activated sorting techniques. The abnormalities were detected by fluorescent in situ hybridation (FISH) with chromosome-specific DNA probes for the detection of trisomy 21, trisomy 18, Klinefelter syndrome 47 XXY, or by polymerase chain reaction (PCR) for the detection of fetal sex, certain Mendelian disorders (as beta-globin mutations), HLA polymorphisms and fetal Rhesus D blood type. However others studies were necessary to determine the sensitivity and specificity of this technique as a noninvasive alternative to conventional methods of prenatal cytogenetic diagnosis. PMID- 11406934 TI - [Prenatal diagnosis of omphalopagus conjoined twins at 13 weeks of amenorrhea]. AB - Conjoined twins are a rare occurrence. We present a case of conjoined twins at 13 weeks' gestation. This case demonstrated the possibility of making an accurate diagnosis of conjoined twins and delineating the extent of organ sharing in the first trimester. This analysis and the discover of another anomalies can help the parents with the option for pregnancy termination. Early diagnosis and precise delineation of the shared organs of conjoined twins are essential for optimal obstetric and postnatal management. PMID- 11406935 TI - [Primary malignant melanoma of the cervix uteri: apropos of 1 case with review of the literature]. AB - The authors report a case of malignant melanoma of the cervix uteri diagnosed in a young 35-year-old patient. Isolated genital bleeding was the beginning symptom. The physical exam showed a cervical dark tumor, with extension into the vagina (superior third), the parametrium was free. Cervical biopsy concluded to a malignant melanoma. No other abnormalities were observed and the tumor was classed stage IIA-FIGO. A total hysterectomy with bilateral annexectomy and pelvic lymphadenectomy was performed. In the way, the patient died after ten months by local and general recurrence. Primitive malignant melanoma of the cervix uteri is rare. Only 39 cases are reported in literature at this day. It is usually initially misdiagnosed mainly in the achromic forms, then the immuno histochemical study is useful. Its treatment is not well codified and its histogenesis was controversial for a long time. PMID- 11406936 TI - [Low genital tract infections]. PMID- 11406937 TI - [TVT or laparoscopic Burch. In favor of TVT ]. PMID- 11406938 TI - [Point of view of laparoscopic Burch]. PMID- 11406939 TI - Are you putting patients in danger? Heed warning on infusion pumps. AB - During on-site surveys in 2001, surveyors will be asking about your use of infusion pumps. The ED is at higher risk for errors and adverse outcomes than other departments because of the types of drugs given and time constraints. Do not use pumps without protection from the free-flow of intravenous fluid/medication into the patient. If your hospital is considering a change in infusion pumps, make sure someone from the ED is involved in the product evaluation. PMID- 11406940 TI - Experts: cut costs of ED observation. AB - Keeping an observation unit operating with current reimbursement might depend on successful implementation of cost-reduction strategies. A "hybrid" model with scheduled-procedure patients in addition to observation patients allows you to staff with fewer nurses per patient. Observe patients with only one specific acute problem who are likely to be discharged within 18 hours. By offering stress testing in close proximity to the observation unit, length of stay is reduced, and the hospital can provide stress testing to inpatients on weekends. PMID- 11406941 TI - Here's how to prevent assaults on staff. AB - ED personnel experience the majority of all hospital assaults, and these attacks can have long-term effects on morale and productivity. OSHA regulations require you to provide staff with a safe working environment. You should have a plan to address security personnel issues, responses to specific scenarios, restraint techniques, and violence prevention. Strategies include flagging violence-prone individuals, using patient liaisons in the waiting areas, and ensuring that ED security officers have been trained to work in hospitals. PMID- 11406942 TI - Medication administration. PMID- 11406943 TI - Use of the PCA Plus II for analgesia. PMID- 11406944 TI - [Development of corneal storage medium--second report. Examination of human cornea]. AB - PURPOSE: To develop and evaluate by histological methods a new corneal storage medium with a simple formula. METHODS: We compared a corneal storage medium which contained minimum essential medium and 2.5% chondroitin sulfate (molecular weight 27,500), pH 7.33, osmolality 320 mOsm/kg with OPTISOL-GS. Paired human donor eyes provided by the Lions Eye Bank of Oregon were stored in a moist chamber until the experiment. A cornea with scleral rim was excised and stored in OPTISOL-GS, and its fellow cornea was stored in the test medium for 5, 10, or 14 days at 4 degrees C. Histological examination of corneal endothelial cells was done by both scanning electron microscopy and transmission electron microscopy. RESULTS: On days 5 and 10, there was no significant difference in histological findings between corneas stored in OPTISOL-GS and those in the test medium. Both corneal groups developed degenerative changes with the increase of storage time, but their histological findings were similar for both storage media. On day 14, corneal endothelial cells showed marked degeneration of intracellular organelles such as a swelling of mitochondria in both media. CONCLUSION: Human corneas stored in the test medium for 14 days maintained their structure as well as those in OPTISOL-GS. This shows that the newly developed corneal storage medium composed can be used for medium-term corneal storage. PMID- 11406945 TI - [Effects of endothelin-1 on choroidal vessels--2. Study using indocyanine green videoangiography]. AB - PURPOSE: The effects of intravenous administration of endothelin-1(ET-1) on choroidal arteries were studied using indocyanine green videoangiography. METHODS: The changes in caliber of the choroidal arteries in eight albino rabbits were examined using indocyanine green videoangiography with a scanning laser ophthalmoscope and an image analysis system. RESULTS: Administration of 10(-9) mol/kg of ET-1 induced a significant reduction in diameter of 50.5-80.1%(median = 64.15%) 5 minutes after administration. The constriction continued for more than 60 minutes. CONCLUSION: This study demonstrates that intravenous administration of ET-1 induces long-lasting constriction of choroidal arteries. This suggests that ET-1 may participate in the regulation of choroidal artery tone. PMID- 11406946 TI - [Effect of caffeine on microcirculation of human ocular fundus]. AB - PURPOSE: To determine the effect of caffeine on the microcirculation in the human ocular fundus. SUBJECTS AND METHODS: The microcirculation of the ocular fundus in 10 eyes of 10 healthy volunteers was studied using a laser speckle tissue circulation analyser. 100 mg of caffeine or placebo was given orally in a double blind test. Square blur rate(SBR), a quantitative index of blood flow velocity, was measured in the temporal site of the optic nerve head(ONH) free of surface vessels and in the middle site of the choroid-retina between the ONH and the macula. Intraocular pressure(IOP), blood pressure(BP), pulse rate(PR), and central critical fusion frequency(CFF) were also measured. These parameters were measured before and for 2 hours after administration. The area under curve(AUC), of the SBR was calculated for each area. Ocular perfusion pressure(OPP) was also obtained from blood pressure and intraocular pressure. RESULTS: The time-course change of SBR value showed much individual difference. Caffeine decreased the AUC of the SBR in the ONH as well as in the choroid-retina significantly. IOP, mean BP, PR, OPP and central CFF did not change significantly. CONCLUSIONS: These results suggest that caffeine may increase blood vessel resistance and decrease blood flow in the ONH and choroid-retina in humans. PMID- 11406947 TI - [Increased iris pigmentation after use of latanoprost in Japanese brown eyes]. AB - PURPOSE: To evaluate the incidence of the increased iris pigmentation in Japanese with topical latanoprost. MATERIAL AND METHODS: One hundred and twelve patients who had homogeneous brown irises were studied. Age of the patients was 23 to 80(average 63.8 +/- 10.4) and follow-up periods were 6 to 12 (10.3 +/- 1.4) months. Slit lamp examination was carefully performed before use and at every month after use of latanoprost. Photographs were taken when necessary. The Kaplan Meier life table method was used to evaluate the incidence of the increased iris pigmentation. RESULT: Iris pigment increase was observed in 47 patients. The incidence of the increased iris pigmentation was calculated as 10.1% at 3 months, 26.4% at 6 months, 50.1% at 9 months and 51.6% at 12 months after use. Pigmentation progressed as a granular concentric or sunrise pattern or a subtype of these. Some pigmentation occurred at one month after use, and progressed monthly. No patients noted their own iris color change, but were told by their relatives. Iris color changes of brown iris were considered to be less strong than in the hazel(green or blue/gray or yellow combined with brown) eye. PMID- 11406948 TI - [Preproliferative diabetic retinopathy sub-classification and glycemic control- relationship between mean hemoglobin A1C value and development of proliferative diabetic retinopathy]. AB - PURPOSE: We evaluated the relationship between long-term glycemic control and the proportion of patients developing proliferative diabetic retinopathy(PDR) in cases of mild preproliferative diabetic retinopathy(PPDR). MATERIALS AND METHODS: We evaluated the relationship between the mean hemoglobin A1C (HbA1C) value during a period of at least 2 years and the proportion of patients developing PDR in cases of mild PPDR, based on our previously proposed subclassification. RESULTS: During follow-up, 27% of all cases developed PDR. The mean HbA1C value in these cases was 9.4%, which was significantly higher than the 7.6% in cases which had not developed PDR. The proportion of patients developing PDR was 48% in cases with a mean HbA1C value 8.6% or more. In contrast, the proportion was 8% in cases with a mean HbA1C value below 8.6%. It was estimated that the proportion of patients developing PDR will approximately double if the mean HbA1C value increases by one percent. The cumulative occurrence rates of PDR at two, 5, and 10 years were estimated to be 5, 28, and 60% in cases with a mean HbA1C value 8.6% or more and 0, 7, and 10% in cases with a mean HbA1C value below 8.6%, respectively. CONCLUSION: Based on the above results, we conclude that more strict systemic and ophthalmological control is indicated for patients with a mean HbA1C value exceeding 8.6%. PMID- 11406949 TI - [Visual evoked potentials elicited by pseudorandom stimulation in macular degeneration]. AB - PURPOSE: To investigate the effect of a central scotoma on the amplitude, latency, and temporal frequency characteristics(TFCs) of the visual evoked potentials(VEPs) elicited by a pseudorandom binary stimulus(PRBS). METHOD: Patients with age-related macular degeneration(AMD) were selected, and VEPs were recorded from 26 eyes with AMD(17 eyes with visual acuity of less than 0.2, and 9 eyes with visual acuity between 0.3 and 0.9). Nine eyes of age-matched normal volunteers served as controls. To acquire the PRBS-VEPs, one eye was stimulated with a PRBS stimulus. The first order kernel was calculated from a cross correlation between PRBS and VEPs. The Fourier transformed first-order kernel was used as the TFC of the VEPs. RESULTS: The P2 latency of the first order kernels was delayed(p < 0.05), and the P2-N2 amplitude was reduced(p < 0.01) in AMD. A depression of the TFC values in the 6-18 Hz band was prominent in the patients with AMD(p < 0.01). CONCLUSION: The TFC, were strongly correlated with the visual acuity of patients with macular degeneration. PMID- 11406950 TI - [Effect of latanoprost on the barrier function of corneal epithelium]. AB - PURPOSE: We evaluated the effect on corneal epithelium barrier function of instillation of prostaglandin F2 alpha ophthalmic solution (latanoprost) for one month. MATERIALS AND METHODS: Ten healthy volunteers and nine glaucoma patients were enrolled in this study. The barrier function was determined as uptake of topically applied sodium fluorescein by the central cornea measured with an anterior fluorophotometer(FL-500, Kowa Co. Ltd). Healthy volunteers and glaucoma patients received 0.005% latanoprost instillation once daily for one month. We measured the uptake of fluorescein by the cornea of each subject before and one month after instillation. RESULTS: Fluorescein uptake before the instillation was 22.2 +/- 16.0 ng/ml (mean +/- standard deviation) and 26.4 +/- 15.1 ng/ml one month after the treatment in the normal group, and it was 55.0 +/- 25.0 ng/ml before treatment and 57.8 +/- 37.0 ng/ml after treatment in the glaucoma group. There was no significant difference in the uptake of fluorescein before and after treatment in either of two groups. CONCLUSION: These results suggested that the barrier function of corneal epithelium was not compromised after the instillation of latanoprost for at least one month. PMID- 11406951 TI - [Pseudo-disc-pits.macular syndrome]. AB - BACKGROUND: Optical coherence tomography(OCT) permits the noninvasive analysis of cross-sectional tomograms of ocular tissue in vivo. CASE: A 67-year-old woman presented with abnormal feeling in the left eye. An ophthalmoscopic examination revealed retinal edema in the macular vascular arcades and radial folds on the fovea. Optic disc pits were not demonstrated ophthalmoscopically or by fluorescein angiography(FAG). OCT disclosed separations in the inner and outer layers of the edematous retina, retinal detachment on the fovea, and cystic schisis(pseudo-pits) beneath the inner limiting membrane of the disc which seemed to be connected with the separation of the inner layers of the retina. CONCLUSION: OCT demonstrated cystic schisis-like pits(pseudo-pits) which were not recognized ophthalmoscopically or by FAG, and the cystic schisis seemed to communicate with the inner separation of the retina. Therefore, we prefer to call such cases pseudo-disc-pits macular syndrome instead of optic disc pit macular syndrome. PMID- 11406952 TI - [A clear cell type meningioma in the upper eyelid ascertained by immunohistochemical examination]. AB - BACKGROUND: We encountered a rare case of clear cell meningioma in the upper eyelid. CASE: A 66-year-old man presented with a slow by progressive, well defined, soft globelar tumor in the left upper eyelid. The tumor showed homogeneous isodensity and was contrasty in a computed tomograph. Histologically, oval-shaped cells with clear cytoplasm had both streamed and whorl configurations, but showed neither psammomas nor calcification. Immunohistochemically, vimentin, epithelial membrane antigen, and protein S-100 were expressed by the tumor cells. CONCLUSIONS: Eyelid meningioma may originate from embryonal remains of the arachnoid in the sheath around the trigeminal nerve, and may vary histologically. Immunohistochemical examination helps to define cases of uncommon subtypes of meningioma. PMID- 11406953 TI - [A case of subacute bacterial endocarditis showing acute macular neuroretinopathy like lesions]. AB - PURPOSE: We report one case of mimic fundus lesions with acute macular neuroretinopathy due to subacute bacterial endocarditis. CASE: A 55-year-old male had about a 1 DD reddish petal-shaped lesion at the level of the retinal pigment epithelium in the macula and a white lesion about 1/6 DD at the level of the retinal pigment epithelium in the upper fovea. Fluorescein angiography showed the reddish lesion to be hypofluorescence due to a filling defect and indocyanine green angiography showed the hypofluorescence was due to a circulatory disturbance of the choriocapillaris. Additionally, we found that there was a severe choroidal circulatory obstruction in the white lesion on the retinal pigment epithelium. After the disappearance of the white lesion, secondary retinal pigment epithelium atrophy remained. CONCLUSION: The macular lesions of acute macular neuroretinopathy were ischaemic lesions of the retinal pigment epithelium formed because of a disturbance at the level of the choriocapillaris. PMID- 11406954 TI - Hospice volunteer training: making the experience more meaningful. AB - Each volunteer coordinator continues to make changes and adjustments in the hospice training classes--some changes arise from circumstances such as staff availability, and some are conscious choices to try new ways of structuring the learning environment. As we shape and refine the experience that we provide for our new volunteers, let us make a special effort to create one that offers not only a comprehensive overview of hospice goals, philosophy, and end-of-life issues, but also a personally challenging and enriching team interaction. PMID- 11406955 TI - Elderly hospice cancer patients' descriptions of their pain experiences. AB - A qualitative research design was used to identify and describe the pain experience of elderly hospice patients with cancer. Eleven participants over the age of 65 receiving hospice services from a for-profit hospice in east Texas were interviewed in their homes. On the basis of a constant-comparative method of analysis, participants identified: (a) multiple sites of pain; (b) hierarchy of pain; and (c) strategies used to decrease pain. Participants differentiated "physical" and "psychological" pain, based on the source of pain. Pain was described as a hierarchy of chronic, acute, and psychological pain, with psychological being the worst. Pharmacological and nonpharmacological strategies were used to decrease their "physical" pain, but participants perceived that there was little they could do about their "psychological" pain. PMID- 11406956 TI - The mouth and palliative care. AB - Oral complications are common among patients with advanced cancer, though relatively little research has been undertaken in this field. This review article discusses the common problem of xerostomia among the terminally ill, together with an overview of oral candidosis, oral viral infections, chemotherapy- and radiotherapy-associated mucositis, and alterations in taste sensation among those with advanced cancer. Suggested management regimes, based on the limited clinical trial data available, are provided where appropriate. PMID- 11406957 TI - Ketamine-fentanyl-midazolam infusion for the control of symptoms in terminal life care. AB - In this report, we describe nine terminally ill patients with metastatic cancer who were treated with an intravenous infusion consisting of ketamine (2 mg/ml)/fentanyl (5 micrograms/ml)/midazolam (0.1 mg/ml) (K/F/M) to control pain after traditional analgesic therapies were unsuccessful. In addition to pain, all patients exhibited some symptoms of cognitive compromise and agitation. After initiation of the K/F/M infusion, all patients exhibited some degree of qualitative improvement in these symptoms as well as in overall pain control. We feel that these observations warrant reporting of the efficacy of this infusion for the treatment of uncontrolled pain and agitation in terminally ill patients when the traditional methods of pain control are inadequate. PMID- 11406958 TI - A collaborative end-of-life care curriculum. AB - Death and dying in America has received limited attention in medical education. The Southern Arizona VA Health Care System and the University of Arizona have collaborated with three nonprofit community hospice programs to develop an end-of life care curriculum. This formal and comprehensive program is offered as a one month elective to senior medical students, residents and fellows. The goal of the program is to improve clinical skills in caring for the dying patient and foster research in palliative and supportive care. PMID- 11406959 TI - Hospice medical directors need ongoing education. PMID- 11406960 TI - A ketamine, fentanyl, and midazolam infusion for uncontrolled terminal pain and agitation. PMID- 11406961 TI - Beyond "death with dignity": a hospice vignette. PMID- 11406962 TI - The personal side of hospice and palliative care education. PMID- 11406963 TI - The needs and supportive networks of the dying: an assessment instrument and mapping procedure for hospice patients. AB - If institutional systems that provide end-of-life care are to survive the demands of managed care, they will need to tie together methods of assessing the needs of the dying, leading to a new understanding of the functioning of a patient's existing helping networks. This paper presents the preliminary findings of a research project conducted at a Midwest hospice. The study utilized a cross sectional correlational survey of patients' needs via the Early Risk and Resiliency Inventory (ERRI), while mapping their helping networks with a Circles of Care Ecomap. It addressed the question, "What is the relationship between the needs of the dying and the formal and informal support provided?" Study findings supported the contention that higher need patients utilized the more expensive institutional services rather than relying on available natural networks; and statistical analysis of the study instrument suggested the creation of a new conceptual domain of psychospiritual need. PMID- 11406964 TI - Factors associated with length of stay in a mid-sized, urban hospice. AB - A recent study by Frantz et al. investigated the relationship between length of stay (LOS) and several factors in a small, rural hospice and found significant differences in LOS by primary physician specialty, referral source, and diagnosis (American Journal of Hospice & Palliative Care, March/April 1999). The purpose of the present study was to replicate and extend the Frantz et al. study in a midsized, urban hospice setting and to examine the relationship of LOS with additional variables, such as living status, discharge status, race, and religion. Significant differences in LOS by gender, diagnosis, physician specialty, referral source, type of insurance, living status, and discharge status were found. No significant differences in LOS were found by race, religion, and place of death. Results are interpreted in the light of previous research findings regarding LOS and in the context of the sample size. Strategies are suggested for increasing patients' LOS. PMID- 11406965 TI - Elements of a comprehensive security solution. PMID- 11406966 TI - Practice management made easy. PMID- 11406967 TI - Giving doctors back their industry. PMID- 11406968 TI - Real-time notification of medication errors. PMID- 11406969 TI - Plan for the worst before disaster strikes. PMID- 11406970 TI - Ready for a shakedown. PMID- 11406971 TI - Coding for dollars. PMID- 11406972 TI - Design, development and delivery: elements of an effective compliance training program. PMID- 11406973 TI - Five ways to manage your data storage. PMID- 11406974 TI - HIPAA: what's hot.... PMID- 11406975 TI - Making disease management patient-centric. PMID- 11406976 TI - What works. Operating system doesn't fail to assist those in need. PMID- 11406977 TI - What works. Survey says: scanning shows satisfaction. PMID- 11406979 TI - Hotlist. Clinical information systems. PMID- 11406978 TI - APC environment poses new compliance risk. PMID- 11406981 TI - What's hot, what's not. PMID- 11406980 TI - APCs: is there a silver lining? PMID- 11406982 TI - Psychiatry, psychiatric rehabilitation, and human values. PMID- 11406983 TI - Personality and recovery: integrating personality assessment data to facilitate the recovery process. AB - The relatively enduring and persistent nature of personality traits means that they will likely continue to impact the course of psychiatric recovery after Axis I symptoms are stabilized. These traits can significantly impact the choices that recovering persons make and the quality of interpersonal relationships with care providers who are trying to facilitate the recovery process. Despite this, they are often inadequately assessed and considered in providing psychiatric care. This manuscript reviews the common combinations of personality traits that have emerged across a variety of clinical samples. The implications of these personality features for the provision of care in an inpatient setting to facilitate recovery are discussed. PMID- 11406984 TI - Restorying psychiatric disability: learning from first person recovery narratives. AB - This qualitative study examines first person accounts of recovery from psychiatric disability. Common themes and patterns are identified and findings are linked to narrative and resiliency theories. Implications for policy, practice, and research are provided. PMID- 11406985 TI - A retrospective study of clubhouse-based transitional employment. AB - This study of clubhouse-based transitional employment (TE) examined the ability of demographic data, diagnosis, and data on TE participation to predict TE tenure and a one-year competitive employment outcome following TE among clubhouse members. Baseline data on 138 club members who participated in TE over a 6-year period were retrospectively retrieved from a computerized database and written records. Most of the jobs held by members were maintenance or production jobs; members' average tenure on TE was 131.26 days. Older members, those with a longer club membership before their last TE job, and those working more days per week had longer average tenure on TE. Average tenure was unrelated to the severity of disability. Forty-two (30.4%) members obtained competitive employment in the one year following their last TE job. Members who worked more total hours on TE were more likely to obtain competitive employment. While aspects of the TE experience predict the move to competitive employment, further studies are needed. PMID- 11406986 TI - Working toward recovery in New Hampshire: a study of modernized vocational rehabilitation from the viewpoint of the consumer. AB - Following the passage of the ADA and the Rehabilitation Amendments of 1992, vocational rehabilitation services offered by New Hampshire community mental health centers have striven collectively to streamline the efficiency of their interventions. Evolved versions of the two most frequently utilized modalities are compared, by measuring vocational satisfaction as it is rated by the perspective of the consumer. Both methods are found to be highly effective, but differ greatly in the nature of their impact and applicability. The strengths and weaknesses of each are identified in the context of psychiatric rehabilitation, and implications are presented for further research. PMID- 11406987 TI - Ethnographic findings from the Washington, D.C., Vocational Services Study. AB - Based on a 20-month period of participant observations and interviews of persons receiving services, employment specialists and clinicians, this ethnographic substudy identified and documented dilemmas encountered during implementation of an assertive, manualized supported employment program, Individual Placement and Support (IPS), situated in a Washington, D.C., community mental health organization that previously focused on clinical interventions but lacked vocational services. Those receiving services, primarily African Americans, had extensive histories of homelessness and dual diagnosis, and minimal work experiences. Real-world issues centered on conflicting expectations and priorities, diverse perceptions of the role of work, and difficulties in integrating vocational rehabilitation with clinical treatment. PMID- 11406988 TI - "Simply to be let in": inclusion as a basis for recovery. AB - This article takes its inspiration from a poem by Borges, in which the author makes a plea to simply be "let in" without being wondered at or required to succeed. Based on the view that these issues have applied historically to people with mental illnesses--first during the period of the asylum, and now more recently as a result of deinstitutionalization--this article argues for the adoption of a broad conceptual framework of inclusion that, based on a disability paradigm, neither alienates or requires people to succeed. First, the ways in which such a framework augments existing approaches to treatment, rehabilitation, and recovery are outlined. Next, the authors describe the three elements of friendship, reciprocity, and hopefulness as aspects of inclusion that may provide a foundation for efforts toward recovery, and illustrate each of these elements through the stories of participants in a supported socialization program. Implications for future research and policy are suggested based on these data. PMID- 11406989 TI - Workplace supports, job performance, and integration outcomes for people with psychiatric disabilities. AB - The notion of natural workplace supports is an essential ingredient in achieving successful supported employment outcomes. This quantitative study examines the characteristics of 243 individuals with psychiatric disabilities participating in supported employment programs in the United States. Diagnoses, psychiatric symptoms, and Global Assessment of Functioning were evaluated in relation to supported employment outcomes, including social interaction, wages, and workplace supports. The results of this study indicate that social interaction and natural workplace supports improve supported employment outcomes for individuals with psychiatric disabilities. PMID- 11406990 TI - Contrary themes on three peer-run warm lines. AB - Peer-run warm lines are relatively new precrisis services, designed for providing social support. Participant observation of three warm lines revealed them to be complex entities, consisting of three contrary themes. Site 1 emphasized connectedness: build peer support networks and establish relationships. Site 2 emphasized nondirectiveness: actively listen and respect boundaries. Site 3 emphasized problem solving: make sure callers are safe for the night. PMID- 11406991 TI - A workshop for receptionists in mental health settings. AB - This article addresses an area which has received little attention in the mental health field--the education and training needs of receptionists. Factors that can assist or impede the contribution of this important position are identified. The authors describe the process used to design and deliver a one-day workshop to enhance receptionists' interpersonal skills, and to increase their understanding of mental health issues. The benefits of this experience for the individual and for mental health organizations are described. PMID- 11406992 TI - The suicidal risk in severe personality disorders: differential diagnosis and treatment. AB - This article describes the clinical approach to patients with severe personality disorders who present suicidal intention and behavior, developed at the Personality Disorders Institute of the Department of Psychiatry at Cornell University Medical College and the Westchester Division of the New York Presbyterian Hospital. It describes the diagnostic evaluation of patients' suicidal potential, personality disorder, and the presence or absence of a spectrum of regressive illness. The analysis of the combined features in these three symptomatic domains determines alternative strategies of psychotherapeutic and psychopharmacological interventions. Within these strategies, transference focused psychotherapy is described as a specific psychodynamic psychotherapy geared to treat characterologically based suicidal and parasuicidal tendencies in the context of the treatment of the patient's personality disorder. PMID- 11406993 TI - One-year follow-up of patients with cluster C personality disorders: a prospective study comparing patients with "pure" and comorbid conditions within cluster C, and "pure" C with "pure" cluster A or B conditions. AB - This one-year, post-treatment prospective study of consecutively admitted patients to a national psychiatric in-patient clinic, compares patients belonging to four subgroups of DSM-III-R personality disorder (PDs): "pure cluster A (N = 21), "pure" B (N = 67), "pure" C (N = 251), and Axis II "comorbid" C (N = 138). Outcome was measured by SCl-90 and occupational status. Axis I disorders were controlled for in all analyses. Contrary to our hypothesis, patients in pure cluster C had no better outcome than either Axis II comorbid cluster C patients or patients with pure cluster A or B. Although pure C patients relapsed in symptom distress after discharge, comorbid C patients did not. C patients with an additional Histrionic PD were less at risk to be a case at follow up (GSI level > 1.00). Cluster C disorders as a whole had negative impact upon outcome in the total sample. These findings suggest the need for better treatment of patients with cluster C conditions. PMID- 11406994 TI - Functional deterioration in individuals with schizophrenia spectrum personality symptoms. AB - Although previous studies have noted functional deterioration in patients with severe schizotypal symptoms who meet criteria for schizotypal personality disorder, we are not aware of any study which examines level of functioning in nonpatients who experience mild schizophrenia spectrum personality (SSP) symptoms. With this issue in mind, occupational functioning was examined in non patient subjects with mild SSP symptoms. Patients were recruited from the community by newspaper advertisements or from the first-degree relatives of patients with schizophrenia. Individuals with no DSM-IIIR Axis I diagnosis and with SSP symptoms (n = 60) and without symptoms (n = 75) participated in the study. The two groups had similar mean age and years of education. Occupational function was evaluated using the Level of Function Scale. The SSP patients with mild symptoms had significantly lower occupational scores (5.62 + 2.50) than the non-SSP patients (7.76 + 0.69; p < 0.001). A total of 39% of SSP patients, compared with 3% of non-SSP patients showed poor occupational functioning (chi 2 = 31, df = 1, p < 0.001). There was a significant deterioration in the socioeconomic status in SSP patients compared with their parents' status. Patients with mild SSP symptoms who were otherwise healthy showed severe difficulties with occupational function. Further research is needed to identify subtle deficits underlying functional deterioration and to develop targets for treatment strategies. PMID- 11406995 TI - Personality disorder in university students: a multitrait-multimethod matrix study. AB - The present study examined the prevalence of personality disorders (PDs) in a sample of first-year university students. The three self-report measures used to assess personality disorders were the (a) Coolidge Axis II Inventory (CATI); (b) Millon Clinical Multiaxial Inventory-II (MCMI-II); and (c) Minnesota Multiphasic Personality Inventory-Personality Disorder Scale (MMPI-PD). The prevalence of PD was estimated by including the number of participants above the cut-off scores selected for each of the three PD instruments. The results indicate prevalence in the range of 0% to 16% for the males, and approximately 1% to 26% for the females. These findings generally confirm the prevalence range of 5% to 15% reported in other investigations of nonpatient samples. With few exceptions, the three tests are generally consistent in their estimates of PD prevalence of all the 11 personality disorders. The multitrait-multimethod matrix reveals that the disorders are generally positively correlated with each other on all the three tests, suggesting considerable construct overlap. Specifically, passive aggressive and schizotypal disorders produce the most convergent correlations. On the MMPI-PD, however, schizoid is a relatively more discriminant disorder. PMID- 11406996 TI - Alexithymia in patients with major depressive disorder and comorbid cluster C personality disorders: a 6-month follow-up study. AB - The authors examined the association between alexithymia, cluster C personality disorders (CPD), and severity of depression among 121 outpatients with major depressive disorder (MDD) in a 6-month, follow-up study. Diagnosis of depression and CPD was confirmed by means of the Structured Clinical Interviews for DSM-III R (SCID I and SCID II). Alexithymia was screened using the 20-item version of the Toronto Alexithymia Scale and severity of depression was assessed using the 21 item Beck Depression Inventory. Results indicated that alexithymic features are common in patients with MDD but often alleviated during recovery from depression. Moreover, comorbid CPD and severity of depression seemed to be associated with poorer recovery from alexithymia. The implications of these findings are discussed. PMID- 11406997 TI - Applicability of personality disorder criteria in late adolescence: internal consistency and criterion overlap 2 years after psychiatric hospitalization. AB - We examined internal consistency and criterion overlap of DSM-III-R personality disorder criteria in late adolescence, 2 years after psychiatric hospitalization. A total of 60 adolescents were assessed with the Personality Disorder Examination. Within-category cohesiveness (internal consistency) was evaluated by coefficient alpha and mean intercriterion correlation (MIC). Between-category criterion overlap was evaluated by examining intercategory mean intercriterion correlations (ICMIC) between all pairs of disorders. Internal consistency was low, with alpha less than .70 for all except borderline and dependent personality disorders. For most disorders, MIC values were higher than ICMIC values. Our findings suggest that personality disorder criteria sets have limited internal consistency in older adolescents. Although the criteria for most personality disorders correlate better with each other than with the criteria for other personality disorders, suggesting some degree of discriminant validity, comparison of these results with a similar analysis performed shortly after hospital admission raises questions about personality disorder construct validity during adolescence. PMID- 11406998 TI - Circular structure of the MCMI-III personality disorder scales. AB - Millon's (1987) circular model of personality disorders was examined in a large sample of psychiatric patients (N = 2,366) who completed the Millon Clinical Multiaxial Inventory-III (MCMI-III; Millon, 1997) as part of routine assessment after presentation for treatment. Principal components analyses were conducted to identify the first two dimensions in MCMI-III base rate scores, weighted and unweighted raw scores, and nonoverlapping scale scores. Similar analyses were made on these scores when acquiescence was partialled out. Circular plots of the scales were examined against Millon's hypothesized arrangement and the model was tested using confirmatory factor analysis. Results replicated those of Strack, Lorr, and Campbell (1990) with the MCMI-II. Millon's horizontal Impassive Expressive dimension was recovered in both regular and residual scores but the vertical axis appeared to represent an Impulsivity-Compulsivity dimension rather than the Autonomous-Enmeshed continuum envisioned by Millon. Although scale order followed Millon's predictions for the most part, a number of departures from theoretical expectations were noted and none of the score sets yielded a good fit to the hypothetical structure. Millon's model appears to have promise as a circumplex that can encompass all of the personality disorders but changes are needed to rectify discrepancies between the theory and empirical findings. PMID- 11406999 TI - The comparative validity of MCMI-II and MMPI-2 personality disorder scales with forensic examinees. AB - This study demonstrated the convergent and discriminant validity of the MMPI-2 and MCMI-II personality disorder scales with forensic examinees. Based on averaged correlational data, the scales performed comparably with previous findings in psychiatric samples. Furthermore, the scales demonstrated increased convergent correlations. Improved convergence was obtained for the Antisocial, Sadistic, Borderline, Schizotypal and Paranoid scales. Decreased convergence on the Dependent and Avoidant scales was also obtained. PMID- 11407000 TI - Incidence of enteroadherence in diarrhoegenic Escherichia coli in infants in Delhi. AB - Fifty-six isolates of Escherichia coli including 40 isolates from diarrhoeic infants and 16 from non-diarrhoeic infants were investigated. Twenty-two of the diarrhoeic isolates were typable, the most common serogroup being 086 (33%). None of the non-diarrheic isolates are typable with EPEC antisera with enteropathogenes. Adherence tests with HEp-2 cell line revealed localized adherence in 23%, diffuse adherence in 14% and aggregative adherence in 5.7% of the 35 isolates tested. Aggregative adherence was not observed in any of the EPEC isolates. None of the isolates in the control group exhibited localized or aggregative adherence. However, 25% of these isolates showed diffuse adherence (DA) which was not significantly different from the incidence of DA (34%) in the test group (p > 0.05). The importance of serogrouping and studying adherence pattern of E. coli isolates in establishing their pathogenic potential is thus emphasized. PMID- 11407001 TI - Occurrence of enterotoxigenic Aeromonas species in foods. AB - Out of a total of 246 food samples of animal origin screened for isolation of Aeromonas spp., 33 (13.41%) were positive for these organisms. Maximum positivity was shown by the samples from fish (28.57%), followed by poultry meat (16.67%), poultry eggs (12.50%), goat meat (12%), buffalo meat (7.69%) and cow milk (5.56%). A. hydrophila was the predominant species (51.52%) followed by A. sobria (39.39%) and A. caviae. Of these, 70.59% A. hydrophila, 69.23% A. sobria and 33.33% A. caviae showed enterotoxigenic reaction in mouse paw oedema test. PMID- 11407002 TI - Dengue/DHF: an emerging disease in India. AB - Dengue/DHF is an emergent disease in India and some parts of country are endemic and periodically contributing annual outbreaks of dengue/DHF. Dengue infection manifests as undifferentiated fever, dengue haemorrhagic fever (DHF) which leads to hospitalization large number of people in a localized area. There is high mortality and morbidity associated with the onset of each dengue outbreak leading to great socio-economic impact. The prevention and control of dengue outbreak depends upon the proper monitoring of the disease case through disease surveillance so as to ensure timely management of cases. Vector surveillance helps in the proper and timely implementation of emergency control measures against dengue vector i.e. Aedes aegypti. There is an urgent need for an effective diagnostic strategy for early diagnosis to shorten the illness duration, hospitalization time and the associated complications. PMID- 11407003 TI - Breeding habitats and larval indices of Aedes aegypti and Ae. albopictus in the residential areas of Calcutta City. AB - Aedes aegypti and Ae. albopictus larvae were found breeding in almost all indoor and outdoor, temporary and/or permanent collections of water, either alone or in association with each other in residential areas of the city, the former throughout the year (June'97-May'98) and the latter during monsoons and post monsoons only. Ae. aegypti showed preference for breeding in chowbachhas, indoors and Ae. albopictus for collections of water in flower pots and discarded containers and outdoors. The larval indices of both the species were highest during monsoons and post-monsoons. PMID- 11407004 TI - Use of locally isolated saprophytic Leptospira strain for serological testing of human leptospirosis. AB - A saprophytic Leptospira isolate recovered from tap water was utilized for serological testing. One hundred-twenty Serum samples comprising 55 cases from PUO/febrile jaundice and 65 samples from apparently healthy individuals were tested by MAT and HA using this environmental saprophytic strain and the results compared with that of Leptospira biflexa semaranga patoc, the standard saprophytic strain commonly employed for sero-diagnosis of leptospirosis. The MAT data showed 96.4 per cent correlation between the two strains. Similarly, the HA results were matching to the extent of 94.5 per cent. Results, therefore, suggest that local saprophytic Leptospira strain may serve as a substitute to serovar patoc for serodiagnosis of leptospirosis. PMID- 11407005 TI - Impact of single round of indoor residual spray with lambda-cyhalotrin 10% WP on Plasmodium falciparum infection in Akola district, Maharashtra State. AB - Lambda-cyhalothrin 10% WP (ICON 10WP) was sprayed from 5th November 1997 at a dose of 25 mg/m2 as indoor residual spray in 74 high risk villages. The spray was completed within 10 days in most of the villages. The monthly entomological monitoring showed nil density of Anopheles culicifacies and Aedes and very low density of non-vector Anopheles and Culex. The impact of Lambda-cyhalothrin spray was discernible right in the month of November 1997 showing 52% reduction in P. falciparum cases as compared to the same month of preceding year. The reduction of P. falciparum cases in three months post-spray period was 77% (from 47 cases to 11 cases) as compared to similar months of preceding year and overall reduction of total malaria cases was 50% during the same period. Since the major part of transmission of P. vivax infection was over by the time Lambda cyhalothrin spray was taken up, obviously the impact on P. vivax infection was not markedly high as compared to P. falciparum infection. Neither cerebral malaria cases nor deaths due to malaria were recorded in the sprayed villages. PMID- 11407006 TI - Levels of CD4 and CD8 among the inhabitants of Manipur, India. AB - Data on mean reference CD4 and CD8% is in general lacking in India. Manipur in the North-East India has high prevalence of HIV infection among the injecting drug users. This study was carried out to establish mean reference CD4 and CD8 cell count in normal and HIV infected individuals in our population, for use in interpretation of these prognostic markers in HIV infected persons. Whole blood sample was collected in EDTA from 14 normal and 23 HIV infected individuals. Fluorescence staining was carried out with FITC conjugated anti-CD4 and CD8 antibodies (Becton Dickinson) directly on whole blood, followed by single step lysis using commercial lysing solution (Optilyse C, Immunotec). The samples were analyzed by two-colour flow cytometry on Coulter Elite cytometer. It was observed that the mean CD4 and CD8 positive cells in normal healthy individuals were 36% (absolute 848/cumm) and 21% (absolute 427/cumm) respectively. The mean CD4% was significantly decreased in HIV infected individuals with a mean value of 13.4% (absolute 246/cumm), while the mean CD8% was significantly increased to 39.2% (absolute 660/cumm) in HIV infected individuals. A lower CD4+ cell count was also observed as compared to the western population among the normal healthy individuals. The mean CD4 and CD8 positive cells in normal healthy adult population were found to be 36% and 21% respectively, and 13.4% and 39.2% in HIV infected individuals respectively. These values should be considered when interpreting CD4 and CD8 counts in HIV infected individuals in this part of the country. PMID- 11407007 TI - An outbreak of Eltor cholera in Aizwal town of Mizoram, India. AB - During the months of May, June and through early part of July 1994, an unusual occurrence of severe dehydrating watery diarrhoea cases and deaths were reported from Aizwal town, the capital of Mizoram, a North-Eastern state of India. Vibrio cholerae 01 biotype Eltor, the causative agent responsible for this outbreak, was isolated from 50.0% of hospitalised cases. The disease affected older children and adults more (52.9%) than younger children below five years of age. Vibrio cholerae 01 strains isolated were uniformly resistant to furazolidone and co trimoxazole, which are commonly advocated in the treatment of cholera specially in children of developing countries. Emergence of such resistant strain is alarming and is of great public health importance. PMID- 11407008 TI - Sexually transmitted infections in long distance truck drivers. AB - A study was undertaken in 670 long distance truck drivers to investigate prevalence of sexually transmitted infections--STI (HIV infection, syphilis, hepatitis-B infection and gonorrhoea) in Nagpur City, Central India. Standard laboratory procedures were followed for carrying out investigations i.e. ELISA for HIV infection, VDRL for syphilis, RPHA for hepatitis-B infection and gram staining for gonorrhoea. A total of 293 (43.7%) subjects had one or more sign/symptoms suggestive of STIs. The prevalence of HIV infection, syphilis, hepatitis-B infection and gonorrhoea was observed to be 15.2%, 21.9%, 5.1% and 6.7% respectively. In conclusion, this study recognized that long distance truck drivers were at high risk for getting STIs including HIV infection. PMID- 11407009 TI - Biochemical and bacteriological study of urinary calculi. AB - Biochemical and bacteriological study of urine, nidus and chemical analysis of 100 calculi from 100 patients admitted in the Regional Institute of Medical Sciences, Imphal from November, 1997 to October 1999 were done. About 47% of the cases had positive urine culture and nidus culture. Escherichia coli was the commonest bacteria isolated both in the urine and nidus of calculi. The commonest radical present in the calculi was calcium while the rarest was uric acid. The stones were composed mainly of calcium oxalate and/or phosphate followed by struvite, then mixed stone. PMID- 11407010 TI - Knowledge, attitude and practice towards malaria in rural tribal communities of south Bastar district of Madhya Pradesh. AB - A sample survey of 100 household heads was carried out in Bastar district to assess villagers knowledge, attitude and practice (KAP) about malaria and their implications for malaria control. Results show that 64.4% of respondents were illiterate and only 20.6% of respondents know the factor for cause of malaria though 45% of household heads received some form of health education from various sources. Respondents who suffered from malaria in previous year constituted 35.8% while 46.8% respondents reported for treatment to nearby PHC, 59.1% respondents admitted the responsibility of male head-of-family to take decision to seek health care malaria. About 34.6% of respondents suspect malaria to any fever. A meagre respondents (8.2%) knew the drug for treatment of malaria, and majority of respondents (70.9%) had taken a complete course of malaria treatment. About 55% respondents did not take any measure to prevent mosquito bite, while 92.4% admitted that they had not taken any measures to prevent malaria. All most all respondents (99.7%) agreed to allow their houses to be sprayed and 99.1% thought that spraying was necessary. The study shows, for successful control of malaria in tribal areas, proper health education about the disease, awareness of personal protection and preventive measures against malaria should be carried out in such a way that should be need based, suitable for the area concern and acceptable, along with active involvement of the communities. PMID- 11407011 TI - Prevalence of HBV and HCV infection among health care workers (HCWs). AB - Hepatitis B virus along with hepatitis C virus infection form a major cause of morbidity and mortality. In order to know the present status in Indira Gandhi Medical College, Shimla, a total of 400 samples were screened from Feb'98 to Jan'99. Two hundred samples from HCWs and the same number of samples from the apparently healthy population which served as the control group were collected. They were screened for the presence of hepatitis B surface antigen (HBsAg) and anti-HCV antibodies by the third generation ELISA. HBsAg was positive in 5% of HCWs while amongst the control group seropositivity was 3.5%. Amongst the HCWs, the laboratory technicians showed highest seropositivity which was 40%. Anti-HCV antibody was not detected in any of the HCWs screened. HCWs form a major risk group for contracting HBV infection. It is therefore of utmost importance to take strict universal precautions and also the need for implementation of immunisation against HBV among HCWs. PMID- 11407012 TI - Malariogenic situations amounting to premunition in areas of Ajodhya Hills of District Purulia, West Bengal. AB - Since a long time, Purulia District in West Bengal has been endemic for malaria. In 1997 and 1998, the district contributed 12.4% (9932 out of 79,811) and 10.0% (13,248 out of 130,288) of malaria cases respectively occurring in West Bengal, resulting 9.45% (7 out of 74) and 5.5% (4 out of 72) of deaths respectively in the state. ABER of the district was 10.4% in 1997 and 8.5% in 1998. PMID- 11407013 TI - Some preliminary observations on the mosquitocidal potential of Mesostoma sp. (Turbellaria: Typhloplaniidae). AB - Observations on the feeding and breeding behaviour of Mesostoma sp. from Delhi are presented. The species is comparable to the M. lingua (a species demonstrated to have high mosquitocidal potential) in predatory potential and may be playing an equally important role here. PMID- 11407014 TI - Plasmodium malariae--a report of three cases. AB - Kasaragod District of Kerala state has never reported cases of Plasmodium malariae. During September 1999-March 2000 a total of 52 slides were reported as positive for P. vivax, P. falciparum and mixed infection. The expert team cross checked these positive slides and three were found positive for P. malariae which were reported as P. vivax. All these had similar clinical features and were either imported cases from endemic areas or local population who visited endemic areas or by persons who came in as construction workers. PMID- 11407015 TI - [Bacterial contamination and presence of Sinorhizobium meliloti in irrigation canal water from the Neuquen River]. AB - A survey of the changes in populations of heterotrophic bacteria, coliform microorganisms and S. meliloti was conducted in samples taken from the water irrigation channels of the Neuquen River (Argentina). Fifty-six water samples were collected during the spring-summer seasons of 1997-1999 years. Both the heterotrophic plate count bacterial and the number of coliforms oscillated between 110-5050 CFU/ml and 8-1400 CFU/100 ml, respectively, during the period this study was carried out. Fecal coliforms were detected in 91.1% of the water samples investigated. Moreover, the results showed that S. meliloti capable of nodulating alfalfa (Medicago sativa L.) Cuf 101 were present in 68% of the water samples and in effectiveness studies, no isolate out of 25 evaluated could be classified as superior N fixers. That is, they did not produce plants equal in weight to nitrate-grown plants (KNO3 0.05%). All the S. meliloti strains were resistant to novobiocin and bacitracin, while 72% of the microsymbionts demonstrated resistance to between seven and ten antibiotics. Results presented in this study showed that irrigation waters of the Neuquen river could act as dispersal agents of both ineffective S. meliloti strains and thermotolerant coliform bacteria. PMID- 11407016 TI - [Expression and characterization of 2 bovine viral diarrhea virus structural proteins (]BVDV)]. AB - The BVDV glycoproteins gp48 and gp53 were expressed in the baculovirus eukaryotic system. Both recombinant proteins were recognized in western blot analysis by monoclonal antibodies and polyclonal serum. Immunofluorescent test demonstrated that gp53 was localized on the cell surface whereas gp48 was in the cytoplasm. The expressed proteins were extracted by non-denaturing detergent treatment. Rabbit antiserum raised against gp53 recombinant protein efficiently neutralized the virus. These results demonstrate that the recombinant proteins have immunological properties similar to those of the native viral protein and that they can be useful as diagnostic reagents. PMID- 11407017 TI - [Detection and genotyping of human papillomavirus (HPV) using PCR-LIS-SSCP]. AB - This study describes a fast and simple method for human papillomavirus (HPV) typing based on the polymerase chain reaction (PCR) amplification of a portion of the viral genome and single strand conformation polymorphism using low ionic strength solutions (LIS-SSCP). PCR products were obtained using My09/My11 and Gp5/Gp6 primers in a nested reaction. The band patterns corresponded to the plasmid HPV clones from HPV-6, -11, -16, -18, -31, -33 and -34. The SSCP minigels were stained with SYBR-Green II. In order to determine diagnostic applicability, 100 cervical samples were studied comprising liquid cytology and paraffin embedded biopsies from patients showing squamous intraepithelial lesions (SILs). The SSCP patterns obtained from the clinical samples and the HPV clones were similar when the same type was present. Therefore, the methodology proved to be efficient and with high reproducibility for the detection and typing of HPV in clinical samples. PMID- 11407018 TI - Influence of bacterial factors on proliferation of bovine mammary epithelial cells. AB - Effects of bovine mastitis pathogen virulence factors on mammary epithelial cell function are not clearly understood. In this study, the effect of streptococcal lipoteichoic acid (LTA), streptokinase, and Escherichia coli lipopolysaccharide (LPS) on proliferation of a primary bovine mammary epithelial cell culture (BTE) and on an established bovine mammary epithelial cell line (MAC-T) was evaluated. Mammary epithelial cells were cultured in the presence of bacterial virulence factors for 48 h at 37 degrees C. BTE cell proliferation was inhibited by streptococcal LTA at 8 and 16 micrograms/ml whereas MAC-T cell proliferation was reduced significantly by concentrations of LTA > or = 2 micrograms/ml. Streptokinase had no effect on proliferation of either MAC-T or BTE cells and LPS inhibited proliferation of BTE but not of MAC-T cells. Effect of LTA and LPS on mammary epithelial cell proliferation could be relevant during the periparturient period when mammary glands are markedly susceptible to new intramammary infection and when mammary epithelial cells undergo extensive proliferation, differentiation and synthesis of milk components. PMID- 11407019 TI - [Inactivation of Trypanosoma cruzi trypanothione reductase by phenothiazine cationic free radicals]. AB - Peroxidase/H2O2/phenothiazine systems produced irreversible inhibition (inactivation) of Trypanosoma cruzi trypanothione reductase (TR). The enzyme inactivation depended on (a) the incubation time of TR with the peroxidase/H2O2/phenothiazine system; (b) the peroxidase nature and (c) the phenothiazine structure. With the more effective peroxidase/H2O2/phenothiazine systems, TR inactivation kinetics presented a relatively fast initial phase, lasting for about 10 min, in which most of the enzyme activity disappeared. This phase was followed by a slower one and, after 30 min incubation, TR was totally inactivated. Three peroxidases were assayed as catalysts of TR inactivation: the horseradish peroxidase (HRP), leukocyte myeloperoxidase (MPO) and modified myoglobin (Mb). Under comparable experimental conditions, the peroxidase system activity decreased in the given order. With HRP systems, 10 microM Thioridazine (TRDZ), Promazine (PZ), Trimeprazine (TMPZ), Prochlorperazine (PCZ), Propionylpromazine (PPZ), Chlorpromazine (CPZ) and Perphenazine (PFZ), produced 95-100% inactivation of TR. With the MPO/H2O2 systems, PZ. TRDZ and TMPZ were the most effective. Under similar experimental condition, the Mb/H2O2/PZ,/TMPZ, /TRDZ and CPZ systems effectively inactivated TR. The presence of alkylamino, piperazinyl, or piperidinyl groups in PTZ N atom (position 10) and -Cl, -CF3, SCH3, COCH2CH3 and -CN in position C2 exerted significant influence on phenothiazine activity. Glutathione (GSH) prevented TR inactivation by the HRP/H2O2/PZ and MPO/H2O2/PZ systems. The HRP/H2O2 and MPO/H2O2/phenothiazines systems generated the corresponding cationic radicals (FTZ.+) the stability of which was limited by their conversion into phenothiazine-sulfoxides (PTZ-SO). The latter ones were inactive on TR. GSH rapidly reacted with PTZ+.; thus producing cation radical detoxication. These reactions fit in well with GSH protection of TR against the peroxidase/H2O2/phenothiazine system, as well as with the FTZ.+ role in phenothiazine cytotoxicity. PMID- 11407020 TI - [Beta-lactam antibiotic sensitivity in Aeromonas spp. of clinical, animal, and environmental origin]. AB - Susceptibility to beta-lactam antibiotics was investigated in Aeromonas spp. Microorganisms were isolated from both, clinical and water creek samples, as well as from processed raw chicken carcasses. Aeromonas like colonies were identified by means of Aerokey II and API 20 E System (Bio-Merieux). A. hydrophila prevailed both of human origin (44%) and water creek samples (41%), while A. caviae ranked first among raw chicken samples (65%). Dilution testing by Agar Method was performed to determine minimum inhibitory concentration (MIC), following NCCLS standards. All tested microorganisms were susceptible to third generation cephalosporin, cefepime, imipenem, aztreonam, and resistant to ampicillin. Only with cefepime and aztreonam exceptions, strains of human origin showed higher values of MIC90 than environmental ones. These results suggest that antibiotic resistance is mainly due to a steady environmental pressure, on account of the widely used above mentioned compounds. PMID- 11407021 TI - [Avian Escherichia coli virulence factors associated with coli septicemia in broiler chickens]. AB - In order to detect phenotypic characteristics associated with pathogenicity, 25 strains of Escherichia coli, isolated from clinical cases of colisepticemia in broiler chickens, were examined to determine the following properties: colicinogenicity, colicin V production, type 1 fimbriae, hemolysin expression and motility. Colicinogenicity occurred in 72% of the strains, 56% of all strains produced colicin V, 84% were positive for type 1 fimbriae and 80% were positive for motility. None of the strains had hemolytic activity; however, all of them, expressed at least one of the other characteristics studied. These results suggest that the diversity of phenotypes detected partially explain the multifactorial nature of avian colisepticemia. PMID- 11407022 TI - [Isolation and characterization of injured coliforms from the drinking water distribution network of La Plata, Argentina]. AB - We screened the La Plata drinking water distribution network for fecal and total coliform bacterial indicator by purification procedures, cultivating 66 membrane filtered samples from the two networks on m-T7 agar. Subterranean and river derived water yielded 13 and 18 confirmed gram-negative bacillus isolates, with 54% and 72% representing total coliforms, respectively. Those from the former source were Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter aerogenes and from the latter Klebsiella oxytoca, Enterobacter agglomerans, and Enterobacter cloacae, genomic group 3. Since 58% of the samples were positive using m-T7 medium it is suggested that the inclusion in standard quality control protocols should be implemented. PMID- 11407023 TI - Designing herbicide formulation characteristics to maximize efficacy and minimize rice injury in paddy environments. AB - Mathematical descriptors, coupled with experimental observations, are used to quantify differential uptake of an experimental herbicide in Japonica and Indica rice (Oryza sativa, non-target) and barnyardgrass (Echinochloa crus-galli, target). Partitioning, degradation, plant uptake and metabolism are described using mass-balance conservation equations in the form of kinetic approximations. Estimated environmental concentrations, governed by the pesticide formulation, are described using superimposed analytical solutions for the one-dimensional diffusion equation in spherical coordinates and by a finite difference representation of the two-dimensional diffusion equation in Cartesian coordinates. Formulation attributes from granules include active ingredient release rates, particle sizes, pesticide loading, and granule spacing. The diffusion model for pesticide transport is coupled with the compartment model to follow the fate and transport of a pesticide from its initial application location to various environmental matrices of interest. Formulation effects, partitioning and degradation in the various environmental matrices, differential plant uptake and metabolism, and dose-response information for plants are accounted for. This novel model provides a mechanism for selecting formulation delivery systems that optimize specific attributes (such as weed control or the therapeutic index) for risk-assessment procedures. In this report we describe how this methodology was used to explore the factors affecting herbicide efficacy and to define an optimal release rate for a granule formulation. PMID- 11407024 TI - Effects of tebuconazole on morphology, structure, cell wall components and trichothecene production of Fusarium culmorum in vitro. AB - The effects of tebuconazole, a systemic fungicide, on the morphology, structure, cell wall components and toxin production of Fusarium culmorum were investigated in vitro. Treatment was by application of four filter paper strips (0.75 cm x 5.0 cm) soaked in 20 micrograms ml-1 fungicide placed around a point inoculum in Petri dishes. Mycelial growth was strongly inhibited by fungicide treatment. Scanning electron microscopic observations showed that the fungicide caused irregular swelling and excessive branching of hyphae. The morphological changes induced by the fungicide at the ultrastructural level included considerable thickening of the hyphal cell walls, excessive septation, the formation of the incomplete septa, extensive vacuolisation, accumulation of lipid bodies and progressing necrosis or degeneration of the hyphal cytoplasm. Non-membrane inclusion bodies were often detected in the hyphal cytoplasm. Furthermore, the formation of new hyphae (daughter hyphae) inside collapsed hyphal cells was common following treatment. The daughter hyphae also displayed severe alterations such as irregular thickening of the cell walls and necrosis of the cytoplasm. Using cytochemical techniques, the labelling densities of chitin and beta-1,3 glucan in the cell walls of the fungicide-treated hyphae were more pronounced than in those of the control hyphae. Moreover, immunogold labelling with antiserum against deoxynivalenol (DON) revealed that Fusarium toxin DON was localized in the cell walls, cytoplasm, mitochondria and vacuoles of the hyphae from the control and the fungicide treatment, but the labelling density in the fungicide-treated hyphae decreased dramatically compared with the control hyphae, indicating that tebuconazole reduced Fusarium toxin production of the fungus. PMID- 11407025 TI - The role of glutathione S-transferases in the detoxification of some organophosphorus insecticides in larvae and pupae of the yellow mealworm, Tenebrio molitor (Coleoptera: Tenebrionidae). AB - The correlation between the natural levels of glutathione S-transferase (GST) and the tolerance to the organophosphorus insecticides parathion-methyl and paraoxon methyl, as well as the interaction of affinity-purified enzyme and the insecticides were investigated in order to collect further information on the role of the glutathione S-transferase system as a mechanism of defence against insecticides in insects. The studies were carried out on the larvae and pupae of the coleopteran Tenebrio molitor L, which exhibit varying natural levels of GST activity. Stage-dependent susceptibility of the insect against insecticides was observed during the first 24 h. However, 48 h after treatment, the KD50 value increased significantly due to the recovery of some individuals. Simultaneous injection of insecticide with compounds which inhibit GST activity in vitro caused an alteration in susceptibility of insects 24 or 48 h post-treatment, depending on stage and insecticide used. Inhibition studies combined with competitive fluorescence spectroscopy revealed that the insecticides probably bind to the active site of the enzyme, thus inhibiting its activity towards 1 chloro-2,4-dinitrobenzene in a competitive manner. High-performance liquid chromatography and gas chromatography revealed that T molitor GST catalyses the conjugation of the insecticides studied to a reduced form of glutathione (GSH). From the above experimental results, it is considered that GST offers a protection against the organophosphorus insecticides studied by active site binding and subsequent conjugation with GSH. PMID- 11407026 TI - Effects of organosilicon pyrethroid-like insecticides on nerve preparations of American cockroaches and crayfish. AB - Quaternary organosilicon pyrethroid-like ethers (five compounds) and alkanes (three compounds) were used for neurophysiological tests. Their activities in inducing repetitive firing in the central nervous cord of the American cockroach (Periplaneta americana) were evaluated by an extracellular recording method. The ethers were more active than the corresponding alkanes. The ability of the compounds to cause conduction blockage was also measured using the same nerve preparations, but the effects were too weak to allow definitive activity values to be determined. The compounds prolonged the sodium tail-current in the crayfish giant axon under voltage clamp conditions. The rate of decay of the tail-current changed in parallel with the reported insecticidal activity against P americana. These findings indicated that tail-current activity was the most useful nerve parameter in predicting insecticidal activity. Regression analysis of the numerical data together with those reported for other alkanes revealed that the higher the tail-current activity, the higher the insecticidal activity when a structural feature and the hydrophobicity were considered separately. The insecticidal activity of the ethers was about seven-fold higher than that of the alkanes with equivalent tail-current activity and hydrophobicity. Variations in insecticidal activity were parabolically correlated with the hydrophobicity [(log P)opt = 9.1] when other factors were similar. PMID- 11407027 TI - Monitoring insecticide resistance in house flies (Diptera: Muscidae) from New York dairies. AB - House flies were collected from dairies across New York state and the levels of resistance to seven insecticides were measured using standard laboratory assays with three to five diagnostic concentrations. The highest levels of resistance were found for tetrachlorvinphos, permethrin and cyfluthrin. Although levels of resistance to methomyl were somewhat lower, they were among the highest ever reported for field-collected house flies. Resistance to pyrethrins was limited primarily to the lowest diagnostic concentration. House flies were susceptible to fipronil at all dairies, suggesting that this material would be highly effective for fly control. The levels of resistance were similar at all the dairies, irrespective of their insecticide use, suggesting substantial movement of flies between facilities. Relative to resistance levels found at New York dairies in 1987, resistance levels had increased for permethrin, were unchanged for tetrachlorvinphos and had decreased for dimethoate. To identify a single diagnostic concentration that could be used in the laboratory assays to assess accurately resistance levels in future studies, we carried out a 'simulated' field bioassay using formulated materials. A diagnostic concentration for each insecticide is proposed on the basis of a comparison of our bioassays. PMID- 11407028 TI - Photolysis of pesticides: influence of epicuticular waxes from Persica laevis DC on the photodegradation in the solid phase of aminocarb, methiocarb and fenthion. AB - Pesticides with N,N-dimethyl and thiomethyl moieties (aminocarb, methiocarb and fenthion) were irradiated under artificial light (lambda > 290 nm) in an amorphous wax phase from Persica laevis DC. The effect of the presence of the wax on the photolysis rate differed in the three pesticides, increasing it in aminocarb, having little effect in methiocarb and slowing it down in fenthion. The presence of the wax affected the qualitative photodegradation behaviour of all the pesticides. The data obtained were compared with those for pirimicarb, which had been studied earlier. PMID- 11407029 TI - Application of coupled liquid chromatography-mass spectrometry in photolysis studies of the herbicide triflusulfuron-methyl. AB - The photochemical fate of the herbicide triflusulfuron-methyl in aqueous solution under UV light (lambda > 290 nm) was investigated. Nine photoproducts were detected and tentatively identified by LC/MS and LC/MS/MS analysis. The main routes of degradation involve the cleavage and/or contraction of the sulfonylurea bridge with the elimination of sulfur dioxide, which increases the acidity of the reaction medium. On the basis of the results obtained a photodegradation pathway is proposed. PMID- 11407030 TI - Mineralization of 2,4-D, mecoprop, isoproturon and terbuthylazine in a chalk aquifer. AB - The potential to mineralize 2,4-dichlorophenoxyacetic acid (2,4-D), mecoprop, isoproturon and terbuthylazine was studied in soil and aquifer chalk sampled at an agricultural field near Aalborg, Denmark. Laboratory microcosms were incubated for 258 days under aerobic conditions at 10 degrees C with soil and chalk from 0.15-4.45 m below the surface. The [ring-U-14C]-labeled herbicides were added to obtain a concentration of 6 micrograms kg-1 and mineralization was measured as evolved [14C]carbon dioxide. The herbicides were readily mineralized in soil from the plough layer, except for terbuthylazine, which was mineralized only to a limited extent. In the chalk, lag periods of at least 40 days were observed, and a maximum of 51%, 33% and 6% of the added 2,4-D, mecoprop and isoproturon, respectively, were recovered as [14C]carbon dioxide. Large variations in both rate and extent of mineralization were observed within replicates in chalk. No mineralization of terbuthylazine in chalk was observed. As a measure of the general metabolic activity towards aromatic compounds, [ring-U-14C]-benzoic acid was included. It was readily mineralized at all depths. PMID- 11407031 TI - Evaluation of uncalibrated preferential flow models against data for isoproturon movement to drains through a heavy clay soil. AB - The uncalibrated predictive ability of four preferential flow models (CRACK-NP, MACRO/MACRO_DB, PLM, SWAT) has been evaluated against point rates of drainflow and associated concentrations of isoproturon from a highly structured and heterogeneous clay soil in the south of England. Data were available for four plots for a number of storm events in each of three successive growing seasons. The mechanistic models CRACK-NP and MACRO generally gave reasonable estimates of drainflow over the three seasons, but under-estimated concentrations of isoproturon over a prolonged period in the first season and over-estimated them in the two remaining seasons. CRACK-NP simulated maximum concentrations of isoproturon over the first two events of each of the three seasons of 156, 527 and 24.4 micrograms litre-1, respectively, and matched the observed data (465, 65.1 and 0.65 micrograms litre-1) slightly better than MACRO (69.1, 566 and 58.5 micrograms litre-1). Automatic selection of parameters from soils information within MACRO_DB reduced the emphasis on preferential flow relative to the stand alone version of MACRO. This gave a poor simulation of isoproturon breakthrough and simulated maximum concentrations were 0, 50.1 and 35.1 micrograms litre-1, respectively. The capacity model PLM gave the best overall simulation of total drainflow for the first two events in each season, but over-estimated concentrations of isoproturon (967, 808 and 51.3 micrograms litre-1). The simple model SWAT represented total drainflow reasonably well and gave the best simulation of maximum isoproturon concentrations (140, 80.2 and 8.2 micrograms litre-1). There was no clear advantage here in using the mechanistic models rather than the simpler models. None of the models tested was able to simulate consistently the data set, and uncalibrated modelling cannot be recommended for such artificially drained heavy clay soils. PMID- 11407032 TI - Fumigant toxicity of volatile natural products from Korean spices and medicinal plants towards the rice weevil, Sitophilus oryzae (L). AB - The fumigant toxicity of various volatile constituents of essential oils extracted from sixteen Korean spices and medicinal plants towards the rice weevil, Sitophilus oryzae L (Coleoptera: Curculionidae), was determined. The most potent toxicity was found in the essential oil from Mentha arvensis L. var piperascens (LC50 = 45.5 microliters litre-1 air). GC-MS analysis of essential oil from M arvensis showed it to be rich in menthol (63.2%), menthone (13.1%) and limonene (1.5%), followed in abundance by beta-pinene (0.7%), alpha-pinene (0.6%) and linalool (0.2%). Treatment of S oryzae with each of these terpenes showed menthone to be most active (LC50 = 12.7 microliters litre-1 air) followed by linalool (LC50 = 39.2 microliters litre-1 air) and alpha-pinene (LC50 = 54.9 microliters litre-1 air). Studies on inhibition of acetylcholinesterase activity of S oryzae showed menthone to have a nine-fold lower inhibitory effect than menthol, despite menthone being 8.1-fold more toxic than menthol to the rice weevil. Different modes of toxicity of these monoterpenes towards S oryzae are discussed. PMID- 11407033 TI - Activity against plant pathogenic fungi of phomalactone isolated from Nigrospora sphaerica. AB - Phomalactone, 5,6-dihydro-5-hydroxy-6-prop-2-enyl-2H-pyran-2-one, produced by the fungus Nigrospora sphaerica, was tested in vitro against nine plant pathogenic fungi, and specifically inhibited the mycelial growth of Phytophthora infestans, with an MIC value of 2.5 mg litre-1. Its inhibitory activities against sporangium and zoospore germination of P infestans were similar to those against Phytophthora capsici. In vivo, at 100 and 500 mg litre-1, it reduced the development of tomato late blight caused by P infestans. PMID- 11407034 TI - Adsorption and degradation of thiazopyr in compost-amended and non-amended soils. AB - Adsorption and degradation of thiazopyr on two unamended soils and a soil amended annually during 8 years with compost were studied under laboratory conditions and compared with the results obtained on soils amended with fresh sewage sludge compost. The adsorption isotherms fitted the Freundlich equation well and a marked sorption increase was found in amended soils. Degradation data followed first-order kinetics and thiazopyr had a half-life of about 75 days at 25 degrees C and 60% water-holding capacity of soil. The addition of fresh compost markedly decreased the rate of thiazopyr degradation, whereas the compost mineralised in the field after annual additions had only a small influence. Incubation studies with sterile soils showed a very significant decrease of the degradation rate, indicating that degradation by micro-organisms was the main pathway of thiazopyr degradation in the soils studied. PMID- 11407035 TI - A new method for assessing foliar uptake of fungicides using Congo Red as a tracer. AB - In order to develop a new method for measuring foliar uptake of fungicides, Congo Red was selected as a tracer, and optimum procedures were established for washing, extracting and analyzing it and fungicides from leaf surfaces. Congo Red, a water-soluble dye, was not absorbed into cucumber or rice leaf, even in the presence of various surfactants, and was completely washable from leaf surfaces by aqueous acetonitrile solutions. Congo Red and fungicides in washings were quantified to calculate the amount of foliar uptake of the latter, by comparing the ratio to Congo Red. The optimum concentration of Congo Red in a formulation should be established in order to minimize its influence on fungicide uptake. Although Congo Red has proved to be useful with a conventional droplet application method, it will give more realistic and practical results with the spraying method used in the present study. PMID- 11407036 TI - [Quality control after hernia operation]. AB - QUESTION: An important feature in surgery is the absence of wound infection after elective sterile operation. That's why the VQG (Verein zur Qualitatssicherung im Gesundheitswesen) from Berne recommends as a standard control the surveillance of wound healing, where hernia repair is suited for a tracer operation [1]. METHOD: In our hospital retrospectively all 106 patients of the year 1999 were asked by a questionnaire, 3 month after operation, if their wound healed primarily or if they noticed any signs of infection or even had to be re-operated again. RESULTS: Most hernias were indirect (71) or direct (11), followed by umbilical hernias (11), epigastric or incisional hernias (4), femoral hernias (2). The majority of procedures were a Shouldice repair (43), followed by mesh implantation according to Lichtenstein (33) or Bassini repair (15). 14 times an umbilical or epigastric hernia was operated by Mayo's method. One Stoppa operation was performed. 7 Hernias were incarcerated at the time of operation. 51 hernias were on the right, 41 on the left side, 5 on both sides. Mean age of patients was 52.9 years; mean duration of hospital stay was 4.42 days. After Shouldice repair hospital stay was 3.9 days with an average patients age of 48.3 years, after Bassini 5.41 days (mean age 46.4 years), after Lichtenstein repair 5.23 days (mean age 66.0 years). 82 (77.4%) from 106 patients returned the questionnaire. 67 (81.7%) were completely satisfied, 11 (13.6%) mentioned minor problems. So 95.1% had a normal primary wound healing without infection. There was no correlation between wound healing, type of hernia, age, hospital stay, surgeon or method of operation. In the same study other complications occurred (e.g. 5% recurrences of hernias, nerve lesions), that could not be integrated into the evaluation because they were excluded by definition. CONCLUSION: Studying the tracer "wound healing after groin hernia repair" allows an approximate follow-up for quality control with little expense. Because of the narrow point of view and the neglect of other complications in the same investigation we estimate the power of the result as limited. PMID- 11407037 TI - [The value of shockwave therapy in treatment of humero-radial epicondylitis]. AB - Numerous therapies exist for the treatment of radial Epicondylitis. A new treatment, Extracorporal Shock-Wave Therapy (ESWT) has recently been proposed. Based on a review of the medical literature various mechanisms of action are presented. Except for the treatment of conditions of the urinary system, ESWT is controversial. Scientific proof of enhanced efficacy of ESWT compared to other treatments of radial Epicondylitis is still lacking. Prospective, randomized follow-up studies of large patient populations under standardized technical conditions are needed. Based on current knowledge, ESWT of radial Epicondylitis should only be applied if three conditions are fulfilled: 1) the diagnosis of radial Epicondylitis has been ascertained, 2) conservative therapies for at least one year failed, and 3) the only alternative is surgery. PMID- 11407038 TI - Intrachannel versus interchannel application of angiogenic factors in transmyocardial laser revascularization. AB - BACKGROUND AND OBJECTIVE: Theoretically myocardial angiogenesis of laser injury can be further enhanced by the addition of angiogenic growth factors. The influence of the way of administration of these factors on vascular growth around the channels is still unclear. MATERIALS AND METHODS: 18 pigs (mean weight 72 +/- 5.2 kg) were randomized to either triads of transmyocardial laser revascularization (TMLR) channels (group 1, n = 6) or isolated channels (group 2, n = 6), or a control group (n = 6). The animals had injections of bovine bone derived growth factor mixture either in the center of the triads in group 1 or within the channels themselves in group 2. Animals were sacrificed one month later for histological analysis. RESULTS: The vascular densities of myocardial areas within the triads of group 1 and around the channels in group 2 were significantly larger than in the control group: 15.2 +/- 3.7/mm2 and 14.2 +/- 3.5/mm2 respectively vs 5.3 +/- 1.6/mm2 (p < 0.001 for both differences). Differences of densities between group 1 and 2 were not statistically significant (p = 0.6). CONCLUSIONS: In this porcine model, the addition of a bovine bone derived growth factor mixture to TMLR significantly stimulates angiogenesis in the areas adjacent to the channels. The place of injection does not influence the angiogenesis intensity. PMID- 11407039 TI - [Endoscopic bursa shaving in acute bursitis]. AB - OBJECTIVES: After resection of olecranon or prepatellar bursa for bursitis there can be long-term skin complications in up to 20%. We wanted to determine the outcome after endoscopic resection, a technique first described in 1990. METHODS: In a period of three years (6/97-7/00) 13 Patients with bursitis were prospectively enclosed in the study and underwent endoscopic resection of the bursa. All patients were examined clinically three weeks and six months postoperatively. RESULTS: In nine patients a resection of the Olecranon bursa was performed, in four patients of the pre-patellar bursa. 11 cases showed a septic bursitis, two a chronic. 50% of the operations were conducted in general anesthesia. We observed no intra- or postoperative complications. The follow-up examinations showed no pain, no recurrence and no loss of joint motion. CONCLUSION: It is our opinion that the endoscopic resection of the Olecranon and pre-patellar bursa for septic bursitis is a promising technique and shows favourable results compared to the open resection. PMID- 11407040 TI - [Osteosynthesis of per- and subtrochanteric fractures by blade plate versus gamma nail. A randomized prospective study]. AB - AIM: To compare clinical and radiological results in per- and subtrochanteric fractures' management with 90 degrees blade plate or Gamma nail fixation; an implant allowing early weight-bearing and fracture healing in correct position remains still difficult. METHOD: Between 1993 and 1995, 26 patients addressed to our Center for a Kyle IV [1] fracture were divided into 2 groups, one fixed with blade plate and the other with Gamma nail. The follow-up for all patients is 12 months. RESULTS: Gamma nail allows early weight-bearing in all patients, fracture healing is acquired at 4.2 months; an operative diaphyseal fracture needed conversion to a long Gamma nail. We observed a slight cut-out that didn't need reoperation. In the blade plate group, we noticed three femoral head necrosis without major flattening, two non-unions, one plate's breakage and two malunions; fracture healing is acquired at 6.3 months. The two non-unions and the plate's breakage didn't need reoperation because of low functional demand. CONCLUSION: We prefer Gamma nail in per- and subtrochanteric femoral fractures' management, it allows early and fast weight-bearing and fracture healing is acquired in all cases. PMID- 11407041 TI - [Intrathoracic goiter--a rare mediastinal tumor]. AB - Intrathoracic, totally ectopic goiters are rare, but have to be considered as a diagnostic possibility in all mediastinal masses. The displacement of the thyroid tissue inferiorly in connection with the embryogenesis of the heart and the large vessels explains the aetiology of the disease. Ectopic goiters are often asymtomatic or oligosymptomatic. They may be classified into two groups according to their location in the anterior or posterior mediastinum. The diagnostic procedures include standard X-ray and CT scan imaging, eventually combined with radionuclide scintigraphy. Once the diagnosis of a intrathoracic goiter is obtained the treatment is surgical (unknown dignity, risk of compression or other symptoms). The approach of choice is through a median sternotomy in case of a anterior location and a posterolateral thoracotomy in case of a posterior location. We discuss the diagnostic and therapeutic approach in 2 patients with mediastinal goiters--one located in the anterior and one in the posterior mediastinum. PMID- 11407042 TI - A large bullet in the bladder. AB - All manner of foreign bodies have been extracted from the bladder. Introduction into the bladder may be through self-insertion, iatrogenic means or migration from adjacent organs. Extraction should be tailored according to the nature of the foreign body and should minimise bladder and urethral trauma. We report a case of a bullet injury to the bladder, which finally presented as a gross hematuria after remaining asymptomatic for four years. We present here an alternative to suprapubic cystostomy with a large bladder foreign body treated via a combined transurethral unroofing followed by removal using a grasper passed through a suprapubic laparoscopic port. PMID- 11407043 TI - [Challenging perioperative anticoagulation in a patient with Budd-Chiari syndrome and heparin-induced thrombocytopenia]. AB - We report on a patient suffering from Budd-Chiari disease who developed heparin induced thrombocytopenia preoperatively. Dorsocranial liver resection and hepatoatrial anastomosis were performed with the extracorporeal circulation and perioperative anticoagulation was achieved with r-hirudin. Surprisingly, thrombus formation was observed in the venous reservoir although intraoperative anticoagulation values were within the targeted level. An additional bolus of hirudin and rinsing the reservoir allowed unproblematic discontinuation of the cardiopulmonary bypass. PMID- 11407044 TI - [Multidisciplinary management of hereditary colorectal cancer]. AB - AIM: The aim of this study was to assess the feasibility and success of multidisciplinary approach for the management of hereditary colorectal cancer. MATERIAL AND METHODS: From November 1998 to November 2000, 32 individuals with putative familial/hereditary predisposition to colorectal cancer were investigated for adenomatous polyposis (attenuated or classical familial adenomatous polyposis coli, FAP) or for hereditary nonpolyposis colorectal cancer (HNPCC). Amsterdam criteria (I and II) and Bethesda guidelines were used to select putative HNPCC kindreds. Clinical data including endoscopy, pathological and operative reports as well as family history were collected. Pre- and post test genetic counseling was offered to at-risk individuals. Genetic testing included microsatellite instability (MSI) and search for germline mutations in the APC, hMSH2 and hMLH1 genes. Immunohistochemistry (IHC) of hMSH2 and hMLH1 protein expression in tumour samples was also performed. RESULTS: 11 APC mutations were characterized, whereas four mutations in HNPCC genes were found in hMSH2 (2) and in hMLH1 (2). MSI and IHC correlated completely for cases with identified pathogenic mutation (100%). CONCLUSION: A thorough evaluation and management of hereditary colorectal requires a multidisciplinary approach. Thus, more mutation carriers can be identified and benefit from appropriate genetic counselling, while non-carrier individuals are relieved from unnecessary surveillance. PMID- 11407045 TI - The health consequences of fuel poverty: what should the role of primary care be? PMID- 11407046 TI - Why inequalities in health matter to primary care. PMID- 11407047 TI - Inequalities in treated heart disease and mental illness in England and Wales, 1994-1998. AB - BACKGROUND: Current government policy is directed towards reducing inequalities in health. General practice is increasingly the focus for health care provision in the United Kingdom and it is therefore important to understand the relationship between socioeconomic factors and health at the general practice level. A deprivation measure was used for the area in which the practice is located and two key priority areas were focused upon: mental health and heart disease. AIM: To explore the relationship between area deprivation and the prevalence of treated heart disease and mental illness in England and Wales. DESIGN OF STUDY: Analysis of data from medical records from practices contributing to the General Practice Research Database. SETTING: A total of 211 practices located in England and Wales. METHOD: The data from the practices were used to examine deprivation differentials in treated disease prevalence rates. RESULTS: There are strong deprivation gradients (highest prevalence in the most deprived category) for treated depression, anxiety, schizophrenia, coronary heart disease and non-insulin-treated diabetes. In general the deprivation differentials are wider in the middle of the age range. Women aged 35 to 64 years in the most deprived areas had rates of treated coronary heart disease two to three times those in the least deprived areas. Men aged 25 to 44 years in the most deprived areas had rates of treated depression 50% higher than those in the least deprived areas. CONCLUSIONS: This study has identified wide deprivation differentials in the prevalence of treated mental illness and heart disease. If all of England and Wales experienced the rates of the least deprived fifth of the population then the number of people being treated for depression, anxiety, and coronary heart disease would fall by 10%, 16%, and 11% respectively. PMID- 11407048 TI - Prevalence of breastfeeding at four months in general practices in south London. AB - BACKGROUND: Successive quinquennial National Infant Feeding Surveys have provided a valuable picture of national and regional variations in infant feeding practices within the United Kingdom. Social variation in breastfeeding has been recognised to be an important source of health inequalities in childhood by the Independent Inquiry into Inequalities in Health Report. AIM: To determine the prevalence of breastfeeding at birth and at four months in a sample of women from urban general practices, its variation between practices, and relation to practice population deprivation scores. To report the timing of introduction of solid feeds. DESIGN OF STUDY: Cross-sectional questionnaire survey based on a random cluster sample. SETTING: Women with infants aged four months in general practices in South London. METHOD: Mode of infant feeding at birth and four months, and time of introduction of solids. Jarman score as a measure of practice population deprivation. Housing tenure, maternal ethnic group, and maternal age at leaving full-time education. RESULTS: Twenty-five general practices were sampled. Median practice Jarman score was 15.0 (interquartile range [IQR] = 12.6 21.9). Responses were received from 1053 out of 1532 mothers approached (69%). Of these, 87% (897) had breastfed at birth, while 59% (609) were still breastfeeding their babies at four months. Mothers in rented accommodation were less likely to breastfeed than owner-occupiers (odds ratio [95% CI] = 0.52 [0.37-0.74]), as were women of white, compared with those of black, ethnic origin (odds ratio [95% CI] = 0.55 [0.36-0.82]). Those who completed up to two years and more than two years education after the age of 16 were 2.94 (95% CI = 1.85-4.66) and 9.25 (95% CI = 6.02-14.21) more likely to breastfeed at four months, respectively, than mothers whose formal education was completed at or before 16 years. Practice-specific rates of breastfeeding ranged from 71% to 100% at birth (median 87%; IQR = 79 93%) and 22% to 83% at four months (median 61%; interquartile range = 47-66%). The intra-practice correlation coefficient for breastfeeding at four months was 0.052 (within-cluster variance = 0.23, between-cluster variance = 0.013). There was no association between breastfeeding at four months and practice-specific Jarman score. Median age of starting solids was 16 weeks (IQR = 15-17 weeks). CONCLUSIONS: Housing tenure, maternal education, and ethnic group are significantly associated with breastfeeding prevalence at four months. Between practice variation in breastfeeding prevalence is not associated with measures of practice population deprivation, as assessed by Jarman scores. Consideration should be given to including information on maternal ethnic group and housing tenure in future National Infant Feeding Surveys. Current weaning practices fall short of the recommendation of the World Health Assembly. PMID- 11407049 TI - Rationale for the new GP deprivation payment scheme in England: effects of moving from electoral ward to enumeration district underprivileged area scores. AB - BACKGROUND: The Department of Health introduced a new deprivation payments system for general practitioners (GPs) on 1 April 1999. Following a three-year phasing in process, registered patients will attract deprivation payments based on the underprivileged area (UPA) score of their enumeration district (ED) of residence, rather than their electoral ward, changing the pattern and distribution of payments throughout England. AIM: To assess the rationale behind the changed deprivation payments system for GPs in England and to examine its impact on GP and practice payments. DESIGN OF STUDY: A quantitative study modelling practice based deprivation payments. SETTING: A total of 25,450 unrestricted principal GPs in 8919 practices in England. METHOD: The effect of three new components in the system were examined: changes in the ED score ranges attracting payment, the percentage increase in the size of successive payment bands, and the total budget. The relationship between consultation rates (used as a proxy for workload) and UPA score was examined, together with changes in GP payments calculated nationally and by geographical area. RESULTS: A total of 11.6% of the population of England live in wards with a UPA score of 30 or more, qualifying for deprivation payments, and a similar proportion (11.4%) live in EDs with a UPA score of 20 or more. The larger percentage increases in the size of payments in successive ED UPA bands is supported by the modelled relationship between consultation rate and UPA score. Financially, under the new deprivations payment system, entitlement widens with 88% of practices receiving a payment. Overall, 74% of GPs gain and 13% lose (3% losing more than 1500 Pounds), with 13% receiving no payment. CONCLUSION: The new ED system maps onto the previous system well. Moreover, it more finely discriminates between smaller areas of different relative deprivation and, thereby, targets payments more accurately. PMID- 11407050 TI - Deprivation, psychological distress, and consultation length in general practice. AB - BACKGROUND: Recent research has shown the benefits of longer consultations in general practice. Approximately 40% of patients presenting to general practitioners (GPs) are psychologically distressed. Studies have shown that psychological morbidity increases with increasing socioeconomic deprivation. The combined effects of psychological morbidity and socioeconomic deprivation on consultation length are unknown. In addition, though it is known that doctors correctly identify half their distressed patients as such, the effect of consultation length on identification is unknown. AIM: To examine factors associated with presentation and recognition of psychological distress in GPs' surgeries and the interaction of these factors with consultation length. DESIGN OF STUDY: A cross-sectional study. SETTING: Nine general practices in the West of Scotland, involving 1075 consultations of 21 full-time GPs. METHOD: The main outcome measures were patient psychological distress (measured by General Health Questionnaire-12), doctors' identification of psychological distress, consultation length, and Carstairs deprivation category scores. RESULTS: The mean consultation length was 8.71 minutes (SD = 4.40) and the prevalence of positive GHQ scores was 44.7%. Increasing GHQ (greater psychological distress) and lower deprivation category scores (greater affluence) were associated with longer consultations. Positive GHQ scoring increased with greater socioeconomic deprivation and also peaked in the 30 to 39 years age group. Recognition of psychological distress was greater in longer consultations (50% increase in consultation length associated with 32% increase in recognition). CONCLUSION: Increasing socioeconomic deprivation is associated with higher prevalence of psychological distress and shorter consultations. This provides further evidence to support Tudor Hart's 'inverse care law' and has implications for the resourcing of primary care in deprived areas. PMID- 11407051 TI - Referral for minor mental illness: a qualitative study. AB - BACKGROUND: Mild depression and anxiety are common problems in general practice. They can be managed by the general practitioner (GP) alone or referred. Previous quantitative studies have shown a large variation between GPs in terms of referral behaviour. The reasons for this variation are not fully understood. AIM: To describe and analyse GP's decision-making processes when considering who should be treating patients with minor mental illness, using a qualitative method. DESIGN OF STUDY: A qualitative interview study. SETTING: Twenty-three GPs in east London and Essex. METHOD: Subjects were chosen using a purposive sampling strategy and participated in one-to-one semi-structured interviews. A grounded theory approach was used for analysis. RESULTS: Two distinct referral strategies were identified--the 'containment' and the 'conduit' approaches. In addition, referrals were found to be of two types--proactive 'referrals to' and reactive 'referrals away'; for minor mental illness the 'referrals away' were found to predominate. Emotive as well as rational responses informed GP decision making on referral. CONCLUSIONS: Explanations of the variation in referral rates need to recognise the emotive responses of individual GPs to minor mental illness. The contribution of guidelines, which assume consistently rational responses to illness, may therefore be limited. PMID- 11407052 TI - Multiple sclerosis masquerade. PMID- 11407053 TI - Women who experience domestic violence and women survivors of childhood sexual abuse: a survey of health professionals' attitudes and clinical practice. AB - Health professionals do not wish to routinely screen women for a history of domestic violence or childhood sexual abuse. However, over 80% believe that these are significant health care issues. Routine screening should not be prioritised until evidence of benefit has been established. PMID- 11407054 TI - Topical antibiotics for acute bacterial conjunctivitis: a systematic review. AB - There has been uncertainty about whether antibiotic therapy confers significant benefit in the treatment of acute bacterial conjunctivitis. This study aimed to assess the efficacy of antibiotic therapy in the management of acute bacterial conjunctivitis. Using standard Cochrane search methods, we identified double blind randomised controlled trials in which any form of antibiotic treatment (topical, systemic or combination) had been compared with placebo in the management of acute bacterial conjunctivitis. Data extraction and analysis followed a pre-defined protocol. Meta-analysis was performed to obtain summary measures of relative risk. Six published trials were identified, of which three fulfilled the eligibility criteria for inclusion in this review. The trials were heterogeneous in terms of their inclusion and exclusion criteria, the nature of the intervention, and the outcome measures assessed. Meta-analysis indicates that acute bacterial conjunctivitis is frequently a self-limiting condition, as clinical remission occurred by days 2 to 5 in 64% (95% confidence interval (CI) = 57-71) of those treated with placebo. Treatment with antibiotics was, however, associated with significantly better rates of clinical remission (days 2 to 5: relative risk (RR) = 1.31, 95% CI = 1.11-1.55), with a suggestion that this benefit was maintained for late clinical remission (days 6 to 10: RR = 1.27, 95% CI = 1.00-1.61). Acute bacterial conjunctivitis is frequently a self-limiting condition but the use of antibiotics is associated with significantly improved rates of early clinical remission, and early and late microbiological remission. Since trials to date have been conducted in selected specialist care patient populations, generalisation of these results to a primary care-based population should be undertaken with a degree of caution. PMID- 11407055 TI - Unequal to the task: deprivation, health and UK general practice at the millennium. AB - The NHS is over 50 years old, but health inequalities remain prevalent in the United Kingdom (UK). Material deprivation may be less apparent; however, social deprivation is becoming worse while the markers of socioeconomic disadvantage remain unsatisfactory. Health is an even more elusive concept; nevertheless, the evidence for an increasing association between deprivation, poor health, and early death is overwhelming. Equally unavoidable is the impact of this social degradation on UK primary care. Service industries have deserted deprived communities but, on the whole, GPs struggle on. Denied the supplementary resources they deserve they become disenchanted, too exhausted to convert incentives into rewards. Clear-headed strategic thinking from the top brass is overdue. PMID- 11407056 TI - Rural deprivation: reflecting reality. AB - In the United Kingdom (UK) there is currently an upsurge of interest in rural affairs. This brings the potential to address some of the gaps in rural health care research. The appropriate description and measurement of rural deprivation is one area consistently identified by UK rural practitioners and policymakers as urgently requiring evidence. Appropriate identification and measurement of deprivation within a rural context is important so that primary care resources can be targeted at those with greatest need. It is believed that current measures of deprivation are inappropriate for rural settings, but relationships between life circumstances and health are only beginning to be addressed by empirical research. In this paper we propose an approach to researching rural deprivation. It is important to be clear about definitions of rurality and deprivation and about the purpose of measurement. The requirement to test a range of indicators for their association with health status and health care need in rural areas and to gather more locally relevant data within primary care settings is highlighted. The relevance, for primary care, of exploring rural deprivation is suggested, along with ideas about a way forward in generating knowledge that can help to characterise and measure rural deprivation in a more sensitive manner. PMID- 11407057 TI - Children and clinical trials. PMID- 11407058 TI - Screening for alcohol abuse. PMID- 11407059 TI - Risk information in general practice. PMID- 11407060 TI - Risk information study was marred by poor questionnaire design. PMID- 11407061 TI - Postal survey responses and questions about income and seeking consent for linkage to medical records. PMID- 11407062 TI - The impact of frequent users of OOH services. PMID- 11407063 TI - Evolutionary ethics--a continuing frustration? PMID- 11407064 TI - [Epidemiology and laboratory diagnostics of legionellae]. AB - The severe pulmonary disease caused by the inhalation of the different Legionella species is called Legionella pneumonia, while the name of the pulmonary disease caused by the most common Legionella (L. pneumophila) is Legionnaires' disease. Another type of disease caused by legionellae is Pontiac fever with influenza like symptoms. Legionella spp. are facultative intracellular parasites. They survive within both monocytes in the human organism and amebae in the environment. To prevent and control the occurrence of legionelloses, legionellae should be surveyed and detected in the environmental (water pipes, air conditioning systems, cooling towers, respiratory equipments, etc.) and clinical (blood, bronchoalveolar lavage, sputum, abscess, etc.) samples. Laboratory diagnosis is complicated by the limitations of the available assays. Thus, it is proposed that the microbiological laboratory diagnosis should be based on the simultaneous application of at least three methods (culturing [on BCYE medium], followed by biochemical assays, serology, molecular biologic methods, such as polymerase chain reaction [PCR], direct demonstration [immunofluorescence microscopy], antigen determination are the most important ones) and on the simultaneous demonstration from three different samples (e.g. lower respiratory tract secretions, sputum, urine, blood culture, serum, moreover, water samples from all potential infectious sources, sediment of hot water tanks, as well as swab samples of faucets and shower heads). The advantage of PCR is that is gives reliable results in one day, in contrast to conventional culturing. However, its sensitivity can not be improved by increasing the sample volume, and neither can it give quantitative results nor can it produce strains for epidemiologic studies, contrary to the method of culturing. It is concluded that PCR and culturing do complement, but do not substitute each other. PMID- 11407065 TI - [Hemorheologic factors in hypertensive and diabetic retinopathy]. AB - Hemorheological factors play an important role in the pathogenesis of different cardiovascular diseases. In the present study the fundus picture and hemorheological parameters (plasma and whole blood viscosity *WBV* by capillary viscosimetry; fibrinogen level; red blood cell aggregation by Myrenne and LORCA aggregometers) of 33 hypertensive patients (23 females, 10 males; mean age: 55 years) and 30 diabetic patients (14 females, 16 males; mean age: 57 years) and healthy controls were examined. The fundus pictures showed different forms of retinopathy in both groups. All the measured hemorheological parameters of the examined patients were in the pathological range and were significantly higher than in healthy controls. In the group of hypertensive patients hematocrit (p < 0.05), plasma fibrinogen (p < 0.01), plasma viscosity (p < 0.01) and whole blood viscosity (p < 0.01) showed a significant increase. The hemorrheological factors showed a parallel deterioration with the severity of hypertension (measured BP values) and of fundus picture, namely their values were significantly (p < 0.01) higher in patients with stage III fundus picture (WBV at 90 1/s: 6.02 mPas), than stage I hypertension (WBV at 90 1/s: 4.51 mPas). In the diabetic group hematocrit (p < 0.01), plasma fibrinogen (p < 0.001), plasma viscosity (p < 0.01), "WBV" (p < 0.001), red blood cell aggregation (p < 0.05) showed a significant increase comparing to healthy controls. These results show that there is a correlation between hemorrheological parameters and fundus picture in hypertensive and diabetic patients, and this suggests that hemorrheological factors may play a role in the development of hypertensive and diabetic retinopathy. PMID- 11407066 TI - [Severe necrotic processes of the penis and scrotum]. AB - Authors summarise the two-decade experience of their department in severe tissue necrosis of penis and scrotum. Presented cases represent a shared field of urology with dermatology, infectology, nephrology and andrology. Tissue necrosis extended to different depth of the penis, from superficial skin and subcutaneous tissues, fascias to the entire organ. Underlying diseases are acute and chronic inflammations, metabolic diseases, vascular lesions. Diagnostic difficulties and therapeutic choices, both conservative and operative, are discussed. PMID- 11407067 TI - [Multiple lymphomatous polyposis--rare case of gastrointestinal hemorrhage]. AB - The authors report a case of a 57 years old male patient, who was admitted to gastroenterology department with upper gastrointestinal haemorrhage. The urgent upper gastrointestinal endoscopy revealed an ulcerated polypoid tumor in the region of angulus of the stomach, and multiple polypoid lesions in the bulbar part of the duodenum. Upon this endoscopic appearance colonoscopy was performed, which revealed a polyposis syndrome in the colorectum. Computer tomography detected mesenterial, retroperitoneal and mediastinal lymph node involvement as well. In this case the primary or secondary origin of the gastrointestinal lymphoma was not verifiable. According to literature data this histological type of the gastrointestinal lymphoma has poor response to chemotherapy, the prognosis is unfavourable. In this particular case the administered chemotherapy resulted in total remission at the lymphoma patient clinically staging III Ae. In the proper follow-up examinations of the patient upper and lower endoscopy, histology samples, laboratory parameters, computer tomography, and physical examination in every 3 months are the methods. PMID- 11407068 TI - [The Imperial and Royal Garrison Hospital in Pest from the Settlement of 1867 until the end of the 19th century]. PMID- 11407069 TI - ["The liver has four vessels" Anatomical knowledge about the liver in the Ebers papyrus]. PMID- 11407070 TI - [Memory of Ferenc Toldy]. PMID- 11407071 TI - [Therapy of neurosyphilis. 1876]. PMID- 11407072 TI - Anticardiolipin antibodies: importance, controversies, discrepancies; the need for guidelines and calibrators. PMID- 11407073 TI - Anticardiolipin antibodies: to be or not to be detectable. PMID- 11407074 TI - The complexity of the mechanisms of action of hyaluronan in joint diseases. PMID- 11407075 TI - Work characteristics, demographic factors and clinical variables could predict work disability in rheumatoid arthritis. PMID- 11407076 TI - Detection of anticardiolipin antibodies in culture supernatants. AB - OBJECTIVE: We have attempted to devise a method for measuring the levels of anticardiolipin antibodies produced in in vitro culture of peripheral blood mononuclear cells isolated from patients with systemic lupus erythematosus (SLE), with or without circulating anticardiolipin antibodies, patients with the primary antiphospholipid syndrome (PAPS) and normal controls. METHODS: Peripheral blood mononuclear cells were isolated and cultured for up to six days, in the presence or absence of added cytokines in the culture medium. Supernatants harvested after culture were tested by ELISA for the presence of anticardiolipin antibodies. RESULTS AND CONCLUSION: Despite variation of the culture conditions and of the cell numbers and populations used, it was found that accurately measurable levels of anticardiolipin antibodies could not be detected reliably in any of the culture supernatants. It is concluded that alternative methods of measurement of antibody production need to be explored. PMID- 11407077 TI - Familial aggregation of polymyalgia rheumatica and giant cell arteritis: genetic and T cell repertoire analysis. AB - OBJECTIVE: Several reports of familial aggregation of giant cell arteritis (GCA) and polymyalgia rheumatica (PMR) have been described although detailed genetic and immunological studies are scarce. Our aims were to investigate the influence of HLA-DRB1 alleles and to analyze the phenotype and T cell receptor (TCR) usage of circulating T lymphocytes in a familial case of GCA and PMR. METHODS: HLA-DRB1 typing was carried out using polymerase chain reaction amplification with specific primers. The study of the circulating T cell repertoire was performed by staining with specific monoclonal antibodies and flow cytometry analysis. RESULTS: Patient 1 developed GCA at the age of 71, four years prior to the diagnosis of PMR in her older brother. The HLA-DRB1 typing of Patient 1 was DRB1*04 (DRB1*0401)/DRB1*12 and in Patient 2 was DRB1*07/DRB1*12. In our patient population, GCA was associated with an increased frequency of HLA-DRB1*04 compared with PMR patients. Regarding T cell phenotype, the brother with active PMR had a higher expression of surface markers indicating activation in both T cell subsets (CD25 and HLA-DR). The sister with GCA showed a pronounced decrease of CD4+/CD45RA+ T cells with respect to her brother with PMR. Both patients carried a significant depletion of CD28 in both subsets, specially within the CD8+ T cell compartment. The BV gene usage differed from one patient to the other. T cell expansions were identified in both patients but the specificities were different. CONCLUSION: We describe an association of GCA and PMR between two first degree relatives with significant genetic and immunologic differences. Our results suggest that the pathogenic mechanisms leading to the development of GCA and PMR are probably multifactorial, and both genetic and environmental factors may contribute to the development of these diseases. PMID- 11407078 TI - Intra-articular hyaluronan injections for the treatment of osteoarthritis of the knee: a randomized, double blind, placebo controlled study. AB - OBJECTIVE: Hyaluronic acid (HA) polymers have been found to be useful as viscosupplements for the treatment of osteoarthritis (OA) in a number of clinical studies. It appears that HA with high molecular weights (HMW) are more effective than low molecular weight HA polymers. METHODS: A single blind, initial randomized study was conducted involving two randomly selected patient groups, which received injections of either placebo or BioHy, a highly purified HMW HA produced by bacterial fermentation. HA was administered intra-articularly and several functional tests, including pain level, stiffness, and physical function, were used to score efficacy at various intervals throughout the study. RESULTS AND CONCLUSION: The results through week 20 indicate that BioHy provides relief for osteoarthritic patients without causing adverse effects, although the study was not sufficiently powered to obtain statistically significant differences between the treatment groups. PMID- 11407079 TI - Evidence for hyaluronan production in the air pouch model in rats. AB - OBJECTIVE: The aim of our work was to investigate the presence of hyaluronan (HA) in the rat air pouch and its behaviour in response to inflammatory stimuli. METHODS: HA levels (by a microplate assay) and the leucocyte count were determined in the fluid obtained from air pouches in which acute or subacute inflammation had been induced by the injection of monosodium urate crystals (MSU) or high density polyethylene (HDPE) debris respectively and in relative controls. RESULTS: In control pouches of both groups, remarkable levels of HA were found; these levels were higher in the very first hours (2475 and 1850 micrograms/l at 6 hrs) and then gradually decreased. In pouches injected with MSU, HA moderately increased (p < 0.001) after 6 hrs, reached a peak after 12 hrs (p < 0.001) and began to taper at 24 hrs (p < 0.001). The leucocyte count was also increased at 6 hrs (p < 0.001), became higher at 12 hrs (p < 0.001) and tapered at 24 hrs (p < 0.001). In the HDPE pouches, HA levels were significantly reduced with respect to controls after 6 hours (p < 0.001), increasing later (p < 0.001) to reach a peak at 24 hrs (p < 0.001), and returning to the original levels, or even below, in the following 72 hours. CONCLUSIONS: These data confirm that the pouch lining produces fair amounts of HA and provide evidence that, in this system, HA levels seem to be influenced by the degree of inflammation even if with variable behaviour in relation to the different characteristics and phases of phlogosis. The present data suggest that the air pouch is a useful experimental model for studies on HA metabolism in either acute or chronic inflammation. PMID- 11407080 TI - Is quality of life affected by season and weather conditions in ankylosing spondylitis? AB - OBJECTIVE: To examine the links between the quality of life (QOL) and season and weather conditions in ankylosing spondylitis (AS) patients. METHODS: A cohort of 146 AS patients (67% males), with a mean age of 47.3 years and a mean disease duration of 12.8 years, answered a self-administered questionnaire, the AS-AIMS2 validated for AS, 4 times over a period of one year in a postal survey. Clinical measures were assessed in a subsample of the cohort. Meterological data including daily temperature, hygrometry, wind speed and atmospheric pressure were collected over the same period. RESULTS: In this cohort, 106 patients completed the follow up. A higher lumbar spine flexibility (Schober index) was associated with a higher climatic temperature and lower wind speed. Physical QOL improved in the summer, as did Social Interaction in the summer and fall, while Role QOL decreased in winter. A lower perceived QOL was significantly associated with a higher temperature and wind speed over the past 4 weeks, and with a higher hygrometry and atmospheric pressure over the past 2 days prior to completion of the questionnaire. CONCLUSION: This study provides some support to the popular belief and frequent patient complaints of a link between health status and perceived quality of life, as assessed by a specific questionnaire, with season and weather conditions. PMID- 11407081 TI - An SV40 large T-antigen immortalized human umbilical vein endothelial cell line for anti-endothelial cell antibody detection. AB - OBJECTIVE: Anti-endothelial cell antibodies in serum of patients with different inflammatory diseases can be detected by a whole cell enzyme-linked immunosorbant assay, using primary cultures of human umbilical vein endothelial cells. To avoid repeated isolation, it would be of great value if an immortal endothelial cell line could be used to perform anti-endothelial cell antibody assays. METHODS: In this study endothelial cells from human umbilical and iliac veins and arteries were transfected with a plasmid containing the Simian Virus 40 large T-antigen. Endothelial cell line(s) derived from this procedure were compared with human umbilical vein endothelial cells in the anti-endothelial cell antibody assay. RESULTS: After transfection, clones of homologous cell populations showed an extended lifespan, before entering a period of crisis. In one human umbilical vein endothelial cell clone a subpopulation of cells escaped crisis and became immortal (EVLC2). Telomerase was activated in this endothelial cell line, resulting in maintenance of the telomere length. There was a significant correlation between anti-endothelial cell antibody testing on human umbilical vein endothelial cells and on the cell line EVLC2. CONCLUSION: The Simian Virus 40 large T-antigen immortalized human umbilical vein endothelial cell line EVLC2 may be useful for the detection of anti-endothelial cell antibodies. PMID- 11407082 TI - Distal extremity swelling with pitting edema in psoriatic arthritis: a case control study. AB - OBJECTIVE: To evaluate the frequency and the clinical characteristics of distal extremity swelling with pitting edema in patients with psoriatic arthritis (PsA). METHODS: This was a case-control study of consecutive outpatients with PsA (old and new diagnosis) observed over a 3-month period in three secondary referral centers in Italy. As controls we used the two consecutive rheumatic outpatients, excluding those with spondylarthropathies, observed after a PsA patient. The demographic and clinical features were assessed by clinical examination and review of the medical records. RESULTS: A total of 183 patients with PsA and 366 controls were evaluated. Distal extremity swelling with pitting edema was recorded in 39/183 (21%) PsA patients and in 18/366 (4.9%) controls (p < 0.0001). In 8/39 (20%) patients this feature presented as a first, isolated manifestation of PsA, and in 8 others it was associated with other features of PsA at diagnosis. The upper and lower extremities were affected, predominantly asymmetrically, in 40% and 60% of the cases respectively. In patients with pitting edema compared to those without this feature, the frequency of Achilles enthesitis and plantar fasciitis, calculated together, was higher (p < 0.05) and the duration of arthritis was significantly lower (p = 0.02). In 7 patients the clinical evidence of a predominant involvement of tenosynovial structures was confirmed by MRI. CONCLUSION: Upper or lower distal extremity swelling with pitting edema due to tenosynovitis, usually unilateral, is a common feature in PsA patients and may represent the first, isolated manifestation of the disease. PMID- 11407083 TI - Estrogen receptor alpha in giant cell arteritis: a molecular genetic study. AB - OBJECTIVE: Giant cell arteritis (GCA) predominantly affects postmenopausal women. Estrogen receptor alpha (ER alpha) accumulates in the cytoplasm of smooth muscle cells, activated mononuclear inflammatory cells and giant cells in the temporal arteries of GCA patients, as well as in smooth muscle cells in arteries from non GCA controls. The aim of this study was to analyse whether this accumulation is related to structural aberrations in the ER alpha mRNA leading to a change in protein structure. METHODS: Total RNA was extracted from inflamed temporal artery tissue in two GCA patients and from non-inflamed arteries in two non-GCA controls. Products from the nested RT-PCR of the cDNA were cloned and plasmid inserts of 20 different clones from each case were investigated using nucleotide sequence analysis. RESULTS: A total of eight different types of transcripts lacking parts of the ER alpha mRNA were detected. Seven of these could be explained by alternative splicing. There were no significant differences between the GCA patients and the non-GCA controls in terms of the number of transcript variants. CONCLUSION: The accumulated cytoplasmic ER alpha in temporal arterial tissue from elderly persons appears mainly to be of wild type. The main structural changes in the ER alpha mRNA may be due to alternative splicing. Somatic mutations of the ER alpha gene appear to be rare and it is therefore unlikely that they are involved in the pathogenesis of GCA. PMID- 11407084 TI - Inhibitory effect of enzyme therapy and combination therapy with cyclosporin A on collagen-induced arthritis. AB - OBJECTIVE: There is increasing interest in the use of combination therapy for rheumatoid arthritis and in the possibility of combining the conventional drug approach with newer antirheumatic therapy. The present study investigates the efficacy of long-term prophylactic enzyme therapy and combination therapy with cyclosporin A in rats with collagen-induced arthritis. METHODS: Rats with collagen-induced arthritis were administered the following drugs: cyclosporin A (5 mg/kg/day and 10 mg/kg/day orally); a mixture of enzymes containing pure substances (bromelain, trypsin, rutin) in the same ratio as in Phlogenzym (PHL, 150 mg/kg, twice daily intrarectally); and a combination of 5 mg/kg/day cyclosporin A plus 300 mg/kg/day PHL for a period of 50 days from the immunization. Levels of serum albumin, serum nitrite/nitrate concentrations, changes in hind paw swelling and bone erosions were measured in the rats as variables of inflammation and destructive arthritis-associated changes. RESULTS: Treatment with 10 mg/kg cyclosporin A, as well as combination therapy with half dosages of cyclosporin A (5 mg/kg) plus PHL significantly inhibited both inflammation and destructive arthritis-associated changes. Significant differences in favor of combination therapy with 5 mg/kg CsA + 300 mg/kg PHL as compared to 5 mg/kg CsA alone were seen in hind paw swelling. Also, reduction of the radiographic scores was more significant in the combination therapy group. Five mg cyclosporin A or PHL alone reduced the disease markers studied to a lesser extent, and in the case of enzyme therapy this occurred at a later stage of arthritis development. CONCLUSION: Our results show the inhibitory effect of enzyme therapy on collagen-induced arthritis in rats, as well as the efficacy of cyclosporin A given in low doses in combination with enzyme therapy, which may be useful in the treatment of rheumatoid arthritis. PMID- 11407085 TI - Intervertebral disc lesions in diffuse idiopathic skeletal hyperostosis (DISH). AB - OBJECTIVE: In order to evaluate the relationships between DISH and vertebral osteochondrosis (degenerative disc disease), the radiographs of the spine of 69 DISH patients were compared to those of 68 controls. METHODS: Radiographs of 69 patients affected by DISH according to Resnick's criteria and of 68 control subjects affected by diseases other than DISH, were evaluated in order to determine the prevalence of vertebral osteochondrosis, diagnosed by the occurrence of moderate to severe reduction in the intervertebral disc height and of the extensive radiographic changes typical of degenerative disc disease, including vacuum phenomena and vertebral body marginal sclerosis. The rate ratios with 95% confidence intervals were computed, with stratification by age groups. RESULTS: Thirty-eight DISH patients (55.1%) and 34 controls (50%) showed vertebral osteochondrosis. Stratification by age revealed an increased prevalence of vertebral osteochondrosis in younger DISH patients with respect to controls (p < 0.05). CONCLUSION: Our results show that vertebral osteochondrosis may be associated with DISH and underline the differences between classification and diagnostic criteria. Moreover, it could be hypothesized that DISH plays a predisposing role in the development of vertebral osteochondrosis during the early stages of the disease, causing an early modification in the physiological curves of the spine. PMID- 11407086 TI - Fatigue and health profile in sicca syndrome of Sjogren's and non-Sjogren's syndrome origin. AB - OBJECTIVE: To assess the health status and fatigue in sicca patients with or without Sjogren's syndrome (SS) and to test whether the immune-inflammatory activity or the extent of the disease predict fatigue in SS. METHODS: The Medical Outcomes Study Short-Form General Health Survey (MOS SF-36) was used in 1 degree SS (n = 90), 2 degrees SS (n = 24), non-SS patients with sicca symptoms (n = 15) and healthy population controls (n = 126). Laboratory values and clinical findings were used to predict fatigue in SS. RESULTS: 74% of the SS and 80% of the non-SS sicca patients felt themselves tired. Vitality score values were 40.2 +/- 20.3 in 1 degree SS, 42.1 +/- 20.6 in 2 degrees SS and 29.0 +/- 15.8 in non SS. The health profiles were similar in 1 degree and 2 degrees SS, worse (p < 0.001) than in normal controls, but in most aspects better than in non-SS sicca patients. In SS neither hemoglobin, ESR nor CRP predicted fatigue. Surprisingly, high serum IgG (p < 0.05), antinuclear antibodies (ANA) (p < 0.01) and SS-A antibodies (p < 0.05) values correlated positively with vitality. The number of disease manifestations correlated negatively with vitality (p < 0.004). The total number of disease manifestations, and ANA and/or SS-A autoantibodies were the best predictors of fatigue, but explained it only to 17-57%. CONCLUSION: Patients with fatigue and perceived ill health but without fibromyalgia had sicca symptoms and low basal tear and salivary secretion rates, indicating that cortical events can lead to a SS-like sicca syndrome. Even in SS fatigue is only in part explained by clinical disease manifestations and laboratory tests assessing inflammation and autoimmunity. Fatigue in both SS and non-SS sicca syndrome more likely correlates to other features, such as neuroendocrine aspects of the disease. PMID- 11407087 TI - T lymphocytes in the synovial fluid of patients with active rheumatoid arthritis display CD134-OX40 surface antigen. AB - OBJECTIVE: To assess the percentage of T lymphocytes, bearing CD134, a member of the TNF receptor superfamily, primarily found on autoreactive CD4+ T cells in the peripheral blood (PB) and synovial fluid (SF) of rheumatoid arthritis (RA) patients. METHODS: The surface expression of CD134 on SF and PB mononuclear cells was performed by flow cytometry in 25 RA patients and correlated to the disease activity. RESULTS: CD134 expression on CD3+, CD4+, CD8+ and CD25+ cells was higher in SF than in PB of RA patients (P < 0.001). No differences were observed in the percentage of CD134+/CD4+ T lymphocytes in the PB of RA patients and controls. Patients with active RA had significantly higher percentage of CD3+/CD134+, CD4+/CD 134+, CD8+/CD134+ and CD25+/CD 134+ than those with inactive disease. CONCLUSION: These findings suggest that CD134+ T cells are involved in the immunopathological process of RA synovitis, maybe mirroring some other autoimmune disease in which autoreactive T cell infiltrating the target tissues largely coexpress CD134. PMID- 11407088 TI - Vascular endothelial growth factor levels in the serum and synovial fluid of patients with rheumatoid arthritis. AB - OBJECTIVE: To determine the vascular endothelial growth factor (VEGF) concentrations in serum and synovial fluid (SF) from patients with rheumatoid arthritis (RA) and to search for relationships between VEGF levels and clinical and laboratory variables. METHODS: We measured VEGF levels using an enzyme-linked immunosorbent assay. Serum samples were obtained from 99 RA patients, 49 osteoarthritis (OA) patients, and 80 normal controls. Paired samples of serum and SF were collected from 32 patients with RA and 15 with OA. RESULTS: The mean serum VEGF concentration was 590.1 pg/ml for RA patients, 286.7 pg/ml for OA patients, and 265.8 pg/ml in controls. The serum VEGF concentration was significantly higher in the RA patients than in the OA patients or the controls (both p < 0.001). Furthermore, the VEGF levels in SF from RA patients were significantly higher than in SF from OA patients (p = 0.017). However, there was no correlation between VEGF levels in serum and SF from the same RA patients. The serum VEGF concentration was correlated with the ESR, serum CRP concentration, serum rheumatoid factor, number of tender and swollen joints, Modified Health Assessment Questionnaire, and patient and physician global assessments of disease activity in RA patients. CONCLUSION: These results suggest that VEGF level is related to RA disease activity, suggesting that VEGF may play some role in the pathogenesis of RA. PMID- 11407089 TI - Mycotic (infected) aneurysm of the popliteal artery and arthritis following Salmonella bacteriemia. AB - We report a case of a mycotic (infected) aneurysm of the popliteal artery due to Salmonella enteritidis. The clinical presentation may be confused with other more common causes of diffuse swollen leg, causing a delay in the diagnosis and proper therapy. Prompt surgical resection of the infected aneurysm together with medical therapy is required for successful treatment. PMID- 11407090 TI - Infliximab in the treatment of adult Still's disease refractory to conventional therapy. AB - In this study we evaluated the efficacy of Infliximab in the treatment of adult Still's disease (ASD) refractory to conventional therapy. Three patients with chronic and active ASD unresponsive to corticosteroids and methotrexate were given intravenous Infliximab infusions at a dosage of 3 mg/kg at weeks 0, 2, 6 and then once every 8 weeks. Methotrexate was maintained in all cases at a dosage of 15 mg/week, whereas the prednisone dose was modified according to disease activity. The follow-up lasted 50 weeks and disease activity improved in all cases during Infliximab therapy. Two patients presented arthralgias and sore throat at 20 and 28 weeks, that was rapidly controlled by Infliximab reinfusion every 4 weeks. One patient relapsed at 18 weeks and dropped out at 22 weeks due to an urticarioid rash after the beginning of the fifth infusion. Infliximab may be effective in the treatment of relapse of ASD refractory to conventional therapy and requiring continuous high dose corticosteroid medication. Further studies are needed to evaluate the long-term safety, efficacy and the optimal schedule of infusion. PMID- 11407091 TI - Hemophagocytosis in autoimmune disease. AB - Reactive hemophagocytic syndrome (HPS) is known to be associated with various autoimmune diseases, as well as infection and/or malignancy. Here we review the features of autoimmune-associated HPS and describe the possible role of autoantibodies, especially antiphospholipid antibodies (aPL), in HPS based on data obtained from our own patients. PMID- 11407092 TI - Activation-induced cell death and Fas-induced apoptosis in patients with systemic or pauciarticular juvenile idiopathic arthritis. AB - OBJECTIVE: To investigate the functionality of the Fas-induced apoptotic pathway in peripheral blood mononuclear cells (PBMC) from patients with systemic or pauciarticular juvenile idiopathic arthritis (JIA). METHODS: PBMC from 12 patients with systemic and 6 with pauciarticular JIA were activated with anti-CD3 and rhIL-2 and then incubated in the presence or absence of the anti-Fas MoAb CH11 inducing activation of the Fas apoptotic pathway. Apoptosis was evaluated by flow cytometry and fluorescence microscopy. RESULTS: The percentage of apoptotic cells following triggering of Fas did not differ between patients with systemic JIA (12.5 +/- 9.5%) or pauciarticular JIA (18.7 +/- 8.9%) and controls (16.1 +/- 6.8%). Evaluation of activation-induced cell death (AICD) in the absence of exogenous triggering of Fas showed that 44% (8/18) of the patients with JIA, compared to none of the controls (0/16), had a percentage of apoptotic cells higher than the mean + 2 SD of controls. The increased AICD was neutralized by the addition of an anti-TNF-alpha antibody. CONCLUSION: Patients with systemic or pauciarticular JIA do not show a defect in the Fas-dependent apoptotic pathway of T cells. The increased AICD present in some patients with JIA appears to be at least in part related to the inflammatory cytokine TNF-alpha. PMID- 11407093 TI - Cervical spine in patients with juvenile chronic arthritis and amyloidosis. AB - OBJECTIVE: To describe cervical spine abnormalities in a group of adult patients with refractory juvenile chronic arthritis (JCA) complicated by secondary amyloidosis (SA). METHODS: The series consists of 49 patients who fulfilled the diagnostic criteria of the European League Against Rheumatism for JCA, here complicated by secondary amyloidosis (SA). We evaluated their clinical records and most recent cervical spine radiographs taken in adult age (> 18 years) at or after the diagnosis of SA. RESULTS: Forty-two (86%) patients evinced inflammatory changes in the cervical spine. Apophyseal joint ankylosis was seen in 31 (63%) and atlantoaxial impaction (AAI) in 28 (57%) patients; anterior atlantoaxial subluxation (aAAS) was noted in 17 (35%) patients, and 19 (39%) had the combination of AAI and apophyseal joint ankylosis. The size of the 4th vertebral body was small or narrow in 14 (29%) patients with JCA onset at a median of 3 years of age (range 1-12). CONCLUSION: Inflammatory cervical spine disorders are common and may be detected along the entire length of the cervical spine in patients with severe refractory JCA. The disorders tend to ankylose the apophyseal joints and destroy the atlantoaxial joints, resulting in aAAS or impaction. These changes will restrict rotatory and bending movements in the cervical spine. A peculiarly small or narrow cervical vertebral body was seen mainly in patients with early onset disease. The present findings shed light on the characteristics and course of the inflammatory changes in the cervical spine in patients with refractory JCA. PMID- 11407094 TI - Severe abdominal involvement as the initial manifestation of cutaneous polyarteritis nodosa in a young girl. AB - We report a young girl who developed ingravescent intestinal symptoms as the first manifestation of cutaneous polyarteritis nodosa (PAN) while the typical skin nodules developed later during the disease course. Cutaneous PAN predominantly affects children and presents with crops of painful skin nodules in the medial aspect of the foot, often preceded by sore throat. Visceral manifestations including gut involvement are commonly associated with the classical form of PAN while they are rarely reported in the cutaneous form. In our patient the severity of the abdominal symptoms required a laparoscopy, which revealed diffuse erythematosus swelling of the intestine on the serosal side. The administration of penicillin and steroids was followed by a dramatic improvement in the disease course. Chronic anterior uveitis developed 4 months after the disease onset and responded to local treatment. At a 2-year follow-up the girl is in good condition under prophylaxis with benzathine-penicillin with no recurrence of the illness. Our case confirms that cutaneous PAN is often related to streptococcal infection, and suggests that ASO titers should be determined in children with vasculitides to ensure a timely diagnosis and treatment of the condition if present. PMID- 11407095 TI - HLA-DRB1 associations in systemic lupus erythematosus patients from northwest Spain. PMID- 11407096 TI - Severe myelotoxicity in rheumatoid arthritis patients treated with oral methotrexate. PMID- 11407097 TI - Acquired Brown's syndrome with juvenile idiopathic arthritis: resolution with early steroid treatment. PMID- 11407099 TI - Gastric Crohn's disease and SAPHO syndrome. PMID- 11407098 TI - Pulmonary embolism after intra-articular injection of methylprednisolone and hyaluronate. PMID- 11407100 TI - Transient hyperthyroidism in a patient with rheumatoid arthritis treated by etanercept. PMID- 11407101 TI - Minor but important symptoms and signs in primary hypertrophic osteoarthropathy. PMID- 11407102 TI - Prostaglandin E1 restores the levels of vWF and ACE in chronic critical limb ischemia in systemic sclerosis. PMID- 11407103 TI - Effects of hemodialysis on advanced bony tophi in a tophaceous gout patient with chronic renal failure. PMID- 11407104 TI - Pilot study using the combination of methotrexate and thalidomide in the treatment of rheumatoid arthritis. PMID- 11407105 TI - Angiotensin receptor blockers and aldosterone antagonists in chronic heart failure. AB - Prognosis in congestive heart failure is directly linked to neurohormonal activation. Angiotensin II through the activation of the renin angiotensin aldosterone system has been the principal focus therapy over the last 2 decades. New agents that target selective blockade of the angiotensin II receptor have been introduced in clinical trials for the treatment of heart failure. Aldosterone has been identified as a critically important neurohormone with direct detrimental effects on the myocardium. Aldosterone antagonists have been used in clinical trials to improve mortality in patients with chronic heart failure. PMID- 11407106 TI - Medical therapy of neurocardiogenic syncope. AB - Neurocardiogenic syncope, alternatively called vasovagal, vasodepressor, or neurally mediated syncope, is a clinical syndrome faced by many clinicians. Its pathophysiology is complicated and not fully understood. Multiple pharmacologic therapies have been evaluated, with no clear ideal agent. Decisions regarding tilt-table testing, selection of pharmacotherapy, and assessment of drug efficacy are not straightforward. This article attempts to assess these issues. PMID- 11407107 TI - Cardiovascular drug-drug interactions. AB - The drug-drug interactions discussed in this article have either documented or suspected clinical relevance for patients with cardiovascular disease and the clinician involved in the care of these patients. Oftentimes, drug-drug interactions are difficult, if not impossible, to predict because of the high degree of interpatient variability in drug disposition. Certain drug-drug interactions, however, may be avoided through knowledge and sound clinical judgment. Every clinician should maintain a working knowledge of reported drug drug interactions and an understanding of basic pharmacokinetic and pharmacodynamic principles to help predict and minimize the incidence and severity of drug-drug interactions. PMID- 11407108 TI - Glycoprotein IIb/IIIa antagonists and low-molecular weight heparin in acute coronary syndromes. AB - The glycoprotein (GP) IIb/IIIa antagonists and the low-molecular weight heparins are the newest additions to the armamentarium of antiplatelet drugs for the treatment of acute coronary syndromes. They are extremely potent inhibitors of platelet aggregation and thrombin generation, respectively. There are currently three GP IIb/IIIa inhibitors (abciximab, eptifibatide, and tirofiban) and two low molecular weight heparins (dalteparin and enoxaparin) approved for use with acute coronary syndromes. Data continue to accumulate outlining the specific roles for these drugs in the treatment of patients with acute coronary syndromes. Clinical trials in patients with acute coronary syndromes have demonstrated that the GP IIb/IIIa antagonists and low-molecular weight heparins offer significant benefit with acceptable safety profiles. Future issues that need to be addressed include refinement of indications for administration and patient selection, comparison between existing agents, evaluation of newer agents, and optimization of dosing to maximize benefit and safety in the use of these powerful new classes of drugs. PMID- 11407109 TI - Beta-blocker therapy in heart failure. AB - beta-blocker therapy in heart failure offers the possibility of arresting or reversing the progressive deterioration in this clinical syndrome. While the mechanism is unclear, improvement in cardiac function has been apparent in virtually every study. Clinical results have shown less consistent improvement. Decrease in hospitalization has been noted, but large-scale clinical trials are underway to assess the effect of beta-blockers on mortality. PMID- 11407110 TI - Update in pharmacologic treatment of hypertension. AB - Initial pharmacologic therapy for hypertension is low-dose thiazide diuretics, beta-blockers, and ACE inhibitors. Increasing data have confirmed that ACE inhibitors have specific benefit in patients with diabetes, atherosclerosis, left ventricular dysfunction, and renal insufficiency. CCBs are alternative agents for ISH in the elderly and appear to decrease stroke with perhaps less protection against progression of renal insufficiency and proteinuria, CAD mortality and new onset heart failure versus other initial agents, especially ACE inhibitors. ARBs are well tolerated and effective blood pressure lowering agents but have not been confirmed as effective as ACE inhibitors for reducing renal progression, clinical events, or mortality from heart failure. Effective pharmacologic antihypertensive therapy may avoid disabling and undetected cerebrovascular disease, cognitive dysfunction, and disturbing symptoms of elevated blood pressure. Vasopeptidase inhibitor, such as omapatrilat, and endothelin-1 antagonist, such as bosentan, may become future agents approved for the reduction of morbidity and mortality with hypertension. The ALLHAT trial continues to examine the potential benefits and harms of amlodipine versus chlorthalidone and lisinopril in a diverse high risk population. Based on ALLHAT data, however, doxazosin is no longer an acceptable initial pharmacological agent. Intensive pharmacologic treatment with blood pressure lowering to less than 130/85 mm Hg is recommended with diabetes, renal insufficiency, and heart failure with additional goal of less than 125/75 mm Hg with renal failure and proteinuria greater than 1 g/24 h, based on multiple outcome studies. PMID- 11407111 TI - Functional versatility in the CRP-FNR superfamily of transcription factors: FNR and FLP. AB - The cAMP receptor protein (CRP; sometimes known as CAP, the catabolite gene activator protein) and the fumarate and nitrate reduction regulator (FNR) of Escherichia coli are founder members of an expanding superfamily of structurally related transcription factors. The archetypal CRP structural fold provides a very versatile mechanism for transducing environmental and metabolic signals to the transcription machinery. It allows different functional specificities at the sensory, DNA-recognition and RNA-polymerase-interaction levels to be 'mixed and matched' in order to create a diverse range of transcription factors tailored to respond to particular physiological conditions. This versatility is clearly illustrated by comparing the properties of the CRP, FNR and FLP (FNR-like protein) regulators. At the sensory level, the basic structural fold has been adapted in FNR and FLP by the acquisition in the N-terminal region of different combinations of cysteine or other residues; which bestow oxygen/redox sensing mechanisms that are poised according to the oxidative stress thresholds affecting the metabolism of specific bacteria. At the DNA-recognition level, discrimination between distinct but related DNA targets is mediated by amino acid sequence modifications in the conserved core contact between the DNA-recognition helix and target DNA. And, at the level of RNA-polymerase-interaction, different combinations of three discrete regions contacting the polymerase (the activating regions) are used for polymerase recruitment and promoting transcription. PMID- 11407112 TI - Environmental sensing mechanisms in Bordetella. AB - The success of a bacterial pathogen may depend on its ability to sense and respond to different environments. This is particularly true of those pathogens whose survival depends on adaptation to different niches both within and outside the host. Members of the genus Bordetella cause infections in humans, other animals and birds. Two closely related species, B. pertussis and B. bronchiseptica, cause respiratory disease and express a similar range of virulence factors during infection, but exhibit different host ranges and responses to environmental change. B. pertussis has no known reservoir other than humans and is assumed to be transmitted directly via aerosol droplets between hosts. B. bronchiseptica, on the other hand, has the potential to survive and grow in the natural environment. Comparison of the manner in which these two organisms respond to external signals has provided important insights into the co ordinate regulation of gene expression as a response to a changing environment. During infection, both species produce a range of virulence factors whose expression is co-ordinated by two members of the two-component family of signal transduction proteins, the bvg (bordetella virulence gene) and ris (regulator of intracellular stress response) loci. When active, the bvg locus directs the activity of a number of virulence determinants in both species whose products, such as adhesins and toxins, establish colonization of the host by the bacteria, although each organism has evolved a slightly different strategy during pathogenesis. B. pertussis, the causative agent of whooping cough, promotes an acute disease and tends to be more virulent than B. bronchiseptica which generally causes chronic and persistent asymptomatic colonization of the respiratory tract. The recently identified ris locus appears to control the expression of factors important for intracellular survival of B. bronchiseptica, but a role for this regulatory locus in B. pertussis infection has not been established. Expression of the virulence determinants controlled by the bvg and ris loci is subject to modulation by different environmental signals, such as low temperature, which act through these two-component systems. Evidence indicates that, for B. bronchiseptica, bvg-controlled determinants expressed under modulating conditions, such as motility, facilitate adaptation and survival in environments outside the host. With B. pertussis, however, there is no apparent requirement for prolonged survival outside the host and this difference is reflected in the expression of different, as yet uncharacterized, determinants as a response to modulating signals. The nature of the gene products involved and their assumed role in the life cycle of B. pertussis remains to be determined. Thus, comparative analysis of these species provides an excellent model for understanding the genetic requirements for pathogenesis of respiratory infection and adaptation to changing environments, both within and outside the host. PMID- 11407113 TI - Microbial metallothioneins. AB - Bacterial metallothioneins bind, sequester and buffer excess intracellular zinc. At present, the vast majority of the available experimental data relate to cyanobacterial metallothionein, SmtA, from Synechococcus PCC 7942. SmtA is required for normal resistance to zinc and smtA-mediated zinc resistance has been used as a selectable marker. The imidazole groups of histidine residues, in addition to the thiol groups of cysteine residues, co-ordinate zinc in bacterial metallothioneins. The structure of bacterial metallothionein must facilitate some discrimination between 'adventitious' and 'adventageous' zinc-binding sites such that under excess zinc conditions metal is predominantly scavenged from the former. It remains unclear whether or not bacterial metallothionein also acts as a zinc store that supplies zinc-requiring proteins or if under some conditions it deactivates a subset of proteins via zinc removal. Expression of smtA is induced in response to elevated concentrations of zinc via the action of SmtB. SmtB has some sequence similarity to the arsenic responsive repressor ArsR and genes encoding related proteins are present in many bacterial genomes. Metal perception by SmtB differs from ArsR. The latter contains a characteristic Cys-Val-Cys motif associated with a DNA-binding helix-turn-helix (the ArsR motif), while the former contains metal-binding motifs associated with a carboxyl-terminal alpha-helix that forms the interface between SmtB dimers (the SmtB motif). Some SmtB-ArsR family proteins, including the zinc sensor ZiaR from the cyanobacterium Synechocystis PCC 6803, have the metal-sensory motifs of both SmtB and ArsR. The mechanisms of action, and the features that allow discrimination between different metal ions by these sensors, are discussed. PMID- 11407114 TI - Extracellular sensing components and extracellular induction component alarmones give early warning against stress in Escherichia coli. AB - The work reported here follows from the proposal that, for efficient induction of numerous extracellular stress responses, cultures contain extracellular stress sensing molecules, termed extracellular sensing components (ESCs). These are directly converted to extracellular induction components (EICs) by stresses, thus providing an early warning system against stress, with very rapid responses occurring on exposure to increasing levels of stress. Although some stress responses appear to involve activation of intracellular sensors, the proposed ESCs and EICs function for many stress tolerance and sensitization responses and for several cross-tolerance and cross-sensitization responses. Because EICs can induce responses in unstressed cells, and because they are small molecules that can diffuse away from the site of formation, they can be considered to be 'alarmones', both warning unstressed organisms of future stress and preparing both stressed and unstressed ones to resist it. Therefore, EICs produced by one group of organisms could affect another group i.e. there could be 'cross-talk' (cell-to-cell communication) with other organisms in an area, to which the EICs diffuse, that has not yet faced the stress. In particular, stimuli that switch on acid tolerance, alkali tolerance, pH sensitization responses and alkylhydroperoxide tolerance are detected by ESCs; these molecules can give rise to EICs in the presence of the stress without organisms needing to be present. Not only does the ESC-EIC interconversion allow rapid switching on of responses, but for some responses it also allows rapid switching off. For some ESCs, the sensor can be modified by the culture conditions, modification leading to altered responsiveness to stress; such sensor changes appear to have evolved to allow the most efficient responses to stress to occur, under defined sets of conditions. In addition, the receptors on the organisms that interact with EICs are modified by culture conditions, so that extracellular components that function as ESCs for some cultures can act as EICs for others. In view of their role in early warning of stress, EICs and ESCs are likely to have important functions in the natural environment, especially in natural waters, in foods and food preparation and production, in hospital, domestic and commercial locations, and in the animal and human body. Findings of major importance relate to the extreme stress tolerance of some EICs. For example, because the acid-tolerance EIC formed at pH 5.0 is a heat-resistant molecule, heat-killed suspensions of acid-tolerant cultures can confer acid tolerance on living E. coli; cultures killed by extreme acidity and alkalinity and by exposure to high levels of UV irradiation or novobiocin are also able to confer acid tolerance on living E. coli. Extracellular components that inhibit induction of stress responses also occur in enterobacteria, since it has been found that AMP and HCO3-, which inhibit acid-tolerance induction, do so by forming extracellular agents that block the functioning of EICs. Similar agents to the above EICs and ESCs may occur in other non-stress-related processes. Systems using these extracellular components are quite distinct in their properties from quorum-sensing systems in Gram-negative bacteria and from those systems that use small peptides in intercellular communication and which induce virulence-related enzyme synthesis in Staphylococcus aureus and competence in streptococci and bacilli. Additionally, probably because the ESCs have evolved to become modified by cultural conditions, the components in the stress-related systems, although relatively small proteins, are much larger than the extracellular components used in the quorum-sensing processes and related systems. It is possible that the extracellular 'protectants' of Nikolaev, which protect E. coli from stress, act similarly to the EICs described here, e.g. by inducing stress tolerance. The antimutagenic factor of Vorobjeva may act similarly, although there is no evidence, so far, to suggest that it acts by inducing tolerance to mutagens. PMID- 11407115 TI - General stress response of Bacillus subtilis and other bacteria. AB - One of the strongest and most noticeable responses of a Bacillus subtilis cell to a range of stress and starvation conditions is the dramatic induction of a large number of general stress proteins. The alternative sigma factor sigma B is responsible for the induction of the genes encoding these general stress proteins that occurs following heat, ethanol, salt or acid stress, or during energy depletion. sigma B was detected more than 20 years ago by Richard Losick and William Haldenwang as the first alternative sigma factor of bacteria, but interest in sigma B declined after it was realized that sigma B is not involved in sporulation. It later turned out that sigma B, whose activity itself is tightly controlled, is absolutely required for the induction of this regulon, not only in B. subtilis, but also in other Gram-positive bacteria. These findings may have been responsible for the recent revival of interest in sigma B. This chapter summarizes the current information on this sigma B response including the latest results on the signal transduction pathways, the structure of the regulon and its physiological role. More than 150 general stress proteins/genes belong to this sigma B regulon, which is believed to provide the non-growing cell with a non specific, multiple and preventive stress resistance. sigma B-dependent stress proteins are involved in non-specific protection against oxidative stress and also protect cells against heat, acid, alkaline or osmotic stress. A cell in the transition from a growing to a non-growing state induced by energy depletion will be equipped with a comprehensive stress resistance machine to protect it against future stress. The protection against oxidative stress may be an essential part of this response. In addition, preloading of cells with sigma B-dependent stress proteins, induced by mild heat or salt stress, will protect cells against a severe, potentially lethal, future stress. Both the specific protection against an acute emerging stress, as well as the non-specific, prospective protection against future stress, are adaptive functions crucial for surviving stress and starvation in nature. We suggest that the sigma B response is one essential component of a survival strategy that ensures survival in a quiescent, vegetative state as an alternative to sporulation. The role of sigma B in related Gram positive bacteria (including cyanobacteria) with special emphasis on pathogenic bacteria is discussed. PMID- 11407116 TI - Microbial molecular chaperones. AB - Protein folding in the cell, long thought to be a spontaneous process, in fact often requires the assistance of molecular chaperones. This is thought to be largely because of the danger of incorrect folding and aggregation of proteins, which is a particular problem in the crowded environment of the cell. Molecular chaperones are involved in numerous processes in bacterial cells, including assisting the folding of newly synthesized proteins, both during and after translation; assisting in protein secretion, preventing aggregation of proteins on heat shock, and repairing proteins that have been damaged or misfolded by stresses such as a heat shock. Within the cell, a balance has to be found between refolding of proteins and their proteolytic degradation, and molecular chaperones play a key role in this. In this review, the evidence for the existence and role of the major cytoplasmic molecular chaperones will be discussed, mainly from the physiological point of view but also in relationship to their known structure, function and mechanism of action. The two major chaperone systems in bacterial cells (as typified by Escherichia coli) are the GroE and DnaK chaperones, and the contrasting roles and mechanisms of these chaperones will be presented. The GroE chaperone machine acts by providing a protected environment in which protein folding of individual protein molecules can proceed, whereas the DnaK chaperones act by binding and protecting exposed regions on unfolded or partially folded protein chains. DnaK chaperones interact with trigger factor in protein translation and with ClpB in reactivating proteins which have become aggregated after heat shock. The nature of the other cytoplasmic chaperones in the cell will also be reviewed, including those for which a clear function has not yet been determined, and those where an in vivo chaperone function has still to be proven, such as the small heat shock proteins IbpA and IbpB. The regulation of expression of the genes of the heat shock response will also be discussed, particularly in the light of the signals that are needed to induce the response. The major signals for induction of the heat shock response are elevated temperature and the presence of unfolded protein within the cell, but these are sensed and transduced differently by different bacteria. The best characterized example is the sigma 32 subunit of RNA polymerase from E. coli, which is both more efficiently translated and also transiently stabilized following heat shock. The DnaK chaperones modulate this effect. However, a more widely conserved system appears to be typified by the HrcA repressor in Bacillus subtilis, the activity of which is modulated by the GroE chaperone machine. Other examples of regulation of molecular chaperones will also be discussed. Finally, the likely future research directions for molecular chaperone biology in the post-genomic era will be briefly evaluated. PMID- 11407117 TI - The future of healthcare delivery: computer-assisted diagnosis, computer-assisted medicine. PMID- 11407118 TI - Coordinating care for the dual-eligible population. AB - Many patients who are eligible for Medicare also are eligible for Medicaid. IDSs that deliver care to these patients, called "dual eligibles," face a challenge of conflicting rules and incentives under existing Federal and state laws governing coverage of these patients. Differences in the Medicare and Medicaid payment rules give IDSs a financial incentive to transfer patients from long-term and postacute to acute care settings in situations in which the patients might receive more appropriate care remaining where they are. Many states are implementing pilot programs to allow IDSs to better coordinate care for dual eligibles. Before states begin implementing pilot or statewide programs, IDSs should be involved in the design of the pilot programs, thereby ensuring that the coordinated funding effectively meets an IDS's requirements for optimum care delivery with adequate payment. PMID- 11407119 TI - The managed care contract: the blueprint for monitoring agreements. AB - Healthcare providers generally undertake monitoring programs of managed care arrangements as a way of analyzing financial performance, uncovering lapses in contractually mandated performance that might expose the organization to financial loss, and gathering information that can be helpful in renegotiating the contract or negotiating new arrangements. To secure access to the information needed to achieve these goals, the provider should ensure that the contract spells out the information required, the health plan's obligations to supply this information, and the consequences of the plan's failure to meet those obligations. Such consequences may include financial penalties for the plan and special termination rights for the provider. Without a contractually explicit assurance that the needed information will be available, a provider may find itself with no way to achieve its contract-monitoring goals. PMID- 11407120 TI - Technology will create a new demand for financial experts. PMID- 11407121 TI - Managing impossible missions: ethical quandaries and ethical solutions. AB - Healthcare financial managers are confronted daily with ethical and practical conflicts, particularly with regard to meeting the conflicting and unduly restrictive laws and regulations with which their organizations must comply. Sometimes the right course of action is not clear, as an examination of the 72 hour rule, safe harbor provisions, and legal interpretations of the False Claims Act reveals. Healthcare financial managers have an ethical duty, based on healthcare codes of ethics and ethical theories, to oppose laws and regulations that put their organizations in untenable legal and ethical situations. PMID- 11407122 TI - CFO compensation reaches record levels. AB - HFMA's 2001 CFO compensation survey finds that CFOs of hospitals and health systems are receiving higher compensation today than ever before. The current average compensation of $127,00--15.5 percent higher than was reported in a similar survey conducted in 1999--is the highest ever recorded by HFMA. Moreover, comparison of the 2001 findings with results of previous surveys shows that the earnings gains for CFOs over the past two years are stronger than they have been at many times in recent history. Factors that were found to influence CFO compensation in 2001 are location, years of service, number of employees reporting to the CFO, supervisory responsibility at the system versus hospital level, experience, and gender. Significant findings of the survey were that the average earnings of CFOs in urban areas are nearly twice those of CFOs in rural areas and that the average difference between earnings of male and female CFOs narrowed from $45,100 in 1999 to $36,800 in 2001. PMID- 11407123 TI - Quality and service are paramount: a survey of healthcare purchasing trends. AB - According to the Sixth Annual Survey Report on Purchasing Value in Health Care, overall health plan premium costs for 2001 increased 10.3 percent for all plan types, with pharmaceutical costs rising at 14.6 percent. The survey shows that 70 percent of employers plan to pass at least some of the increased costs on to employees, but only 14 percent plan to reduce or eliminate coverage for certain services. Both providers and employers are more likely than are health plans to believe that cost pressures are hurting quality. Employers say they generally are satisfied with the quality of care their employees receive through their plans. Access and quality of care influence employers' health plan purchasing decisions more than cost, but health plans greatly underestimate the importance employers place on quality of care and effective administration. Most employers plan to increase use of the Internet for a broad range of activities, including annual enrollment, health and wellness promotion, and plan administration. PMID- 11407124 TI - Consumer focus can spur group practice turnaround. AB - Many healthcare organizations have lost money on their employed group practices. The solution to this dilemma is not necessarily divestment of the group practices. Instead, some healthcare organizations should view their physicians as an asset. Healthcare organizations and physicians need to develop a new framework for their relationship to optimize their competitive advantage. Three guiding principles that will help accomplish this objective are to recast the healthcare organization-physician relationship to focus on the consumer, reconfigure the economic model to exceed consumer demands, and restructure the group practice to encourage fiscal and service excellence. In developing a new relationship framework, the stakeholders need to define the group practice's mission, strategic direction, composition, infrastructure, compensation model, and structure. PMID- 11407125 TI - Without action, prescription benefit could bankrupt Medicare. PMID- 11407126 TI - Coping strategies used in residential hospice settings: findings from a national study. AB - The purpose of this study was to explore professional caregivers' coping strategies for dealing with the deaths of patients in residential hospices in the United States. Using the Guide to the Nation's Hospices, 1996-97, purely residential hospices were identified and invited to participate in the study. Employees at each residential hospice were asked to complete the Ways of Coping Questionnaire. Results indicated that positive reappraisal coping was the most frequently used coping strategy. Employees dissatisfied with the coping experience reported greater use of confrontive coping, escape-avoidance coping, and accepting responsibility strategies. The findings suggest that in-service training related to coping strategies and environmental interventions may help in strengthening the coping responses of residential hospice staff. PMID- 11407127 TI - Anesthesia practitioner involvement, invasive treatments, and need in hospice pain management: a survey of patient care coordinators. AB - Pain management is one of the major concerns for the terminal patient. The hospice care team is a highly trained group of health care providers in the area of symptom control, including pain management, for the dying patient. Anesthesia providers also specialize in pain control. The purposes of this study were to survey hospice patient care coordinators to gain an understanding of anesthesia practitioners' involvement with hospice patients, hospice patients' access to anesthesia pain management services, and hospice patient care coordinators' attitudes toward the necessity of anesthesia pain management services for the hospice community. A questionnaire was developed to assess these issues. In general, the findings reflected minimal anesthesia practitioner involvement in the hospice community. Fifty-two percent reported that patients could benefit from invasive treatments offered by anesthesia practitioners. Forty percent responded that more patients could be considered as candidates for invasive pain management techniques if procedures were performed in the patient's home or hospice. Access to anesthesia pain management services was limited by distance to pain clinics and anesthesia practitioners, and more anesthesia pain management services were needed for hospice patients in smaller communities. Cost of anesthesia pain management was frequently proposed as a prohibitive factor. PMID- 11407128 TI - A longitudinal study of pain in hospice and pre-hospice patients. AB - Pain continues to be a very formidable foe in the care of the hospice patient. The incidence among hospice admissions may range from 50 to 80 percent. With such a high initial incidence of pain, the rapidity with which pain can be controlled becomes a very high priority for the hospice effort. The assessment and management of pain in a home-based hospice program presents some unique problems- and opportunities, in that much of this work is done by hospice nurses on site, rather than by the physician, who might remain quite removed from the process. In the study described below, 250 consecutive admissions to either a hospice, or pre hospice (bridge) program were assessed for pain on admission. Those with pain scores of 5 or greater (on a 1 to 10 scale) were followed daily for 15 days by phone to reassess pain and treatment effects. Of the 250 consecutive patients surveyed, 41 (16 percent) gave pain scores of 5 or greater. Mean pain scores for the 41 patients dropped to < 5 within 24 hours of admission. PMID- 11407129 TI - Olanzapine: another psychotropic? AB - Olanzapine is a favorable drug with a potential treatment of delirium, psychotic depression, mood disorders and nausea with a single drug in advanced cancer patients. Additional benefits include weight gain. The medication has few drug interactions, a narrow therapeutic window and high drug cost, which is a disadvantage compounded by a single route of administration. Reduced hospitalizations, at least in the psychiatric patient, may offset the drug cost. PMID- 11407130 TI - The role of tamoxifen in hospice: pros and cons. PMID- 11407131 TI - Drawing the line. PMID- 11407132 TI - Screening for depression in palliative care. PMID- 11407133 TI - Palliative care in amyotrophic lateral sclerosis: a new tool in the fight against an old enemy. PMID- 11407134 TI - Equianalgesic dosing of transdermal fentanyl. PMID- 11407135 TI - Pain is under-reported in nursing home patients. PMID- 11407136 TI - HCFA issues draft of Minimum Data Set for palliative care. PMID- 11407137 TI - End-of-life care listed as a national research priority. PMID- 11407138 TI - Treatment options to manage pain at the end of life. AB - Experts believe that the time preceding death can be comfortable if people die without pain, with dignity, and in their own way. Given current analgesic options, psychological and spiritual interventions, and an effective health care delivery system, all these goals are achievable. Pain management is one of the most important aspects of end-of-life care. Effective analgesics should be chosen carefully, in keeping with the patient's overall condition, level and stability of pain, and specific patient/family wishes. Ideally, analgesics should be initiated as soon as appropriate. The variety of routes of delivery, ranging from oral to transdermal or epidural to intrathecal, allows a selection that will achieve comfort and yet be least troublesome for the caregiver and patient. As the palliative care specialty continues to grow in the United States, it is imperative that health care professionals in the field develop basic to advanced primers to assist in equipping colleagues in all specialties with an understanding of effective opioid use, as well as the multidimensional aspects of helping patients achieve comfort at the end of life. PMID- 11407139 TI - Psychoanalysis and medication: is real integration possible? AB - The author first summarizes historical tensions surrounding the use of medication in psychoanalytic treatments. Next he examines the now-popular viewpoint that clinicians should consider only descriptive criteria in deciding whether to medicate. He then describes how medication use can potentially undermine patients' full integration of psychic experiences. Finally, he presents clinical material to illustrate why clinicians ought to consider patients' defensive processes as a core part of the initial decision about medication use. PMID- 11407140 TI - Beyond dualism: psychoanalysis and medication in the 21st century. AB - Dualistic thinking continues to pervade psychoanalytic views of psychopathology. The author points out the shortcomings of this perspective as represented in Mark Swoiskin's article, "Psychoanalysis and Medication: Is Real Integration Possible?" She contends instead that the latest scientific evidence does not support a dichotomy between organic (biological) and psychodynamic (nonbiological) factors in psychopathology. She presents two cases to illustrate both the challenge and the importance of reformulating psychoanalytic views in an integrated, holistic way. PMID- 11407141 TI - Further thoughts on dualism, science, and the use of medication in psychoanalysis. AB - In response to Deborah Cabaniss's article, "Beyond Dualism: Psychoanalysis and Medication in the 21st Century," the author further considers the differences between the aims of symptom reduction and psychic integration, the concept of mind-body dualism, and the nature of scientific inquiry as they pertain to the use of medication in psychoanalytic therapies. He warns against the collapsing of concepts, aided by a misapplication of science, with respect to how we listen to, organize, and respond to clinical material. He argues that only when such scrutiny occurs can the important and challenging questions pertaining to the use of medication in psychoanalytic therapies be meaningfully considered. PMID- 11407142 TI - Safe sex and first-episode schizophrenia. AB - The need for educating patients about the dangers of unprotected sexual activity is well documented in the literature. Using clinical examples, the authors describe safe-sex strategies for patients experiencing their first episode of schizophrenia. Interventions are based on a 2-year experience of working in a hospital-based treatment and research project with 68 patients. Strategies that begin in the healing phase of schizophrenia take place in both individual and group sessions. First-episode patients are encouraged to speak explicitly about their sex-related behaviors, and HIV testing is suggested when needed. The goal of this approach is to emphasize safe-sex/HIV prevention strategies within a framework of good clinical practice. PMID- 11407143 TI - A note on transference and alliance: I. Transference--variations on a theme. AB - Conceptual and pragmatic difficulties are encountered in relating and differentiating transference from alliance. Transference and alliance, along with the real relationship, are component elements of the analytic relationship, and are mutually involved in intermingling and interaction at all points of the analytic process. Variants of transference are discussed with an eye to distinguishing their differentiation from and relationship to alliance components and functions. Forms of transference differentiated are classical transferences (libidinal and aggressive), transference neuroses, transference psychoses, narcissistic transferences, selfobject transferences, transitional relatedness, transferences as psychic reality, and relational or intersubjective transferences. Transference mechanisms--specifically displacement, projection, and projective identification--and their role in transference development are discussed. Differences in the concept of transference conceived classically as opposed to relationally or intersubjectively are explored. Therapeutic advantages and limits of these differentiations are considered. PMID- 11407144 TI - A note on transference and alliance: II. Patterns of interaction. AB - Conceptual and pragmatic difficulties are encountered in relating and differentiating transference from alliance. Transference and alliance, along with the real relationship, are component elements of the analytic relationship, and are mutually involved in intermingling and interaction at all points of the analytic process. Variants of transference, as discussed in a companion article (Meissner, 2001), are differentiated from and related to alliance components and functions. Transference and alliance can interact in oppositional terms, each undermining or obfuscating the other; an emphasis on alliance may subvert transference and, conversely, transference may act as one of the major sources of misalliance. They may also sustain and reinforce each other; alliance often serves as a vehicle for providing a safe context for emergence of difficult transference derivatives, and benign and mild forms of positive or idealizing transference can offer reinforcement to the analytic relationship, and in some degree, to the alliance. Some forms of transference, especially selfobject transferences, may enjoy significant overlap with alliance functions. Therapeutic implications and limits of these differentiations are considered. PMID- 11407145 TI - How was this story told in the mother tongue? An integrative perspective. AB - Most of the psychoanalytic literature on the mother tongue has focused on the clinical therapeutic encounter. The authors applied findings from therapy research using the Thematic Apperception Test (TAT) to tease out differences and similarities in how stories are told in the mother tongue versus the acquired language. This projective test provides a rich medium to explore the individual's inner world, fantasies, thoughts, wishes, sense of self, and self-in-relation-to other via a narrative that is spontaneously produced in response to standardized stimuli. The TAT was administered to four participants in both the mother tongue and the acquired language. The narratives were then analyzed using two different rating systems. Differences in sense of self, self-in-relation-to-other, and level of pathology were identified using one participant's narratives. Although no global generalizations can be made from one study, the authors suggest that people tell their story differently when speaking in the mother tongue versus the acquired language. PMID- 11407146 TI - Morphometrical study on senile larynx. AB - The aim of the study was a morphometrical macroscopic evaluation of senile larynges, according to its usefulness in ORL diagnostic and operational methods. Larynx preparations were taken from cadavers of both sexes, of age 65 and over, about 24 hours after death. Clinically important laryngeal diameters were collected using common morphometrical methods. A few body features were also being gathered. Computer statistical methods were used in data assessment, including basic statistics and linear correlations between diameters and between diameters and body features. The data presented in the study may be very helpful in evaluation of diagnostic methods. It may also help in selection of right operational tool' sizes, the most appropriate operational technique choice, preoperative preparations and designing and building virtual and plastic models for physicians' training. PMID- 11407147 TI - The neuronal structure of the dorsal lateral geniculate nucleus in the guinea pig: Golgi and Kluver-Barrera studies. AB - On the basis of Golgi and Kluver-Barrera preparations we have distinguished four types of neurons in the dorsal lateral geniculate nucleus of the guinea pig: 1. Fusiform neurons with 1-3 thick dendritic trunks arising from each pole of the soma. The dendritic trunks branch twice dichotomically. The branches sometimes show varicosities. 2. Pear-shaped cells. From one pole of the perikaryon one or two thick dendritic trunks arise, from the opposite pole an axon emerges. The ends of the dendritic branches divide in a tuft-like manner (a characteristic feature of the interneurons). 3. Rounded neurons with 4-7 dendritic trunks without cones. The dendritic trunks branch once or twice dichotomically and give finally 2-3 thin ramifications which show a varicose course and knob-like protuberances. 4. Triangular cells with 3 thick, chronically arising dendritic trunks. They bifurcate dichotomically. The surface of the dendritic trunks and of their branches is smooth. PMID- 11407148 TI - A comparative study on oncostatin M secretion by transplantable melanoma cells with regard to their biological properties and progression. AB - The authors have studied oncostatin M secretory activity of cells of two lines of transplantable melanoma with different biological properties. The level of OSM in melanoma cells supernatant was determined by ELISA test and OSM bioactivity was measured by using biotest line sensitive to OSM cytostatic activity--A 375 melanoma cells. OSM presence was confirmed in cellular lysates by western blot. It has been demonstrated that a spontaneous alteration of the transplantable melanotic melanoma line into an amelanotic line connected with tumour progression was accompanied by an increase of OSM release. PMID- 11407149 TI - The levels of TNF-alpha in cerebrospinal fluid and serum do not correlate with the counts of the white blood cells in acute phase of ischaemic stroke. AB - Stroke-induced inflammatory reaction, which leads to invasion of leukocytes into the evolving brain infarct, seems to play a key role in the deterioration of brain ischaemic impairment. We have studied CSF and serum levels of tumour necrosis factor-alpha (TNF-alpha), the potent proinflammatory cytokine, and peripheral white blood cells (WBC) counts in patients within the first 24 hours of ischaemic stroke. TNF-alpha levels in CSF and serum as well as WBC counts were increased. There was no correlation between TNF-alpha levels either in CSF and serum or in WBC counts. The results of our study suggest that increased CSF TNF alpha levels may represent acute intracerebral inflammation in stroke, whereas elevated levels of TNF-alpha in serum may reflect the peripheral proinflammatory state as well as stroke-induced systemic inflammatory reaction. Increased CSF and serum TNF-alpha levels do not correlate with the elevation of WBC counts, suggesting that TNF-alpha overexpression observed in early phase of stroke is not dependent on increased total number of peripheral leukocytes. PMID- 11407150 TI - A variation in the formation of the median nerve: communicating branch between the musculocutaneous and median nerves in man. AB - We encountered variation in the formation of the median nerve in a 66-year-old male cadaver during dissection of the upper extremity of 20 adult cadavers. The dissections were made at the Department of Cellular Biology and Anatomy, Louisiana State University Medical Center. The median nerve was formed by fusion of four branches, three of them coming from the lateral cord and one from the medial cord. The normal radix from the lateral cord followed a very close oblique course over the axillary artery. The first unusual radix to the median nerve had an anastomoses from the musculocutaneous nerve to the median nerve in the proximal part of the left arm. The second unusual radix also came from the musculocutaneous nerve after it had pierced the coracobrachialis muscle and then joined with the median nerve. These kinds of variations are vulnerable to damage in radical neck dissection and other surgical operations of the axilla and upper arm. The communicating branch can be explained on the basis of its embryologic development and also ought to be distinguished from the other nerve variations in the upper extremity. The aim of this paper is to provide additional information for the classification of previously found communications between the musculocutaneous and median nerves. PMID- 11407151 TI - Impact of educational intervention on knowledge of mothers regarding home management of diarrhoea. AB - A pre and post comparison study was carried out in the field practice area of M.S. Ramaiah Medical College Bangalore, Karnataka to assess the impact of educational intervention on the knowledge of mothers of under five children on home management of diarrhoeal diseases. Sample of 225 mothers were included in the study. The study was conducted in 3 stages. Stage I--initial knowledge, attitude and practice of mothers was assessed. Stage II--one to one educational intervention was conducted and supported by audiovisual aids and live demonstration. Stage III--included post intervention knowledge, attitude and practice after 2 months and 2 years. After the educational intervention, there was significant improvement on knowledge of mothers regarding definition of diarrhoea (P < 0.001), signs of dehydration (P < 0.001), awareness of ORS solution (P < 0.001), correct preparation of ORS solution (P < 0.001), shelf-life of ORS solution (P < 0.001), seeking health care (P < 0.001) and rational drug therapy during diarrhoea (P < 0.001). McNemar test was used to find out the change in knowledge before and after the educational intervention. The overall knowledge scores improved significantly after 2 months (P < 0.001) as well as 2 years (P < 0.001) of the educational intervention. Though the proportion of mothers retaining the knowledge at the end of 2 years dropped, yet there was significant improvement (P < 0.001) when compared to the baseline study. PMID- 11407152 TI - Dietary supplies of iodine and thiocyanate in the etiology of endemic goiter in Tripura. AB - In the post-salt iodization phase, a study on iodine nutriture status was conducted in Tripura of North East India. The clinical variable of the study was goiter and the biochemical variables were urinary iodine and thiocyanate. Random sampling methodology was followed for selecting the study areas in the State. In each study area, the studied population consisted of school children of both sexes in the age group 6-15 years. The total study areas were 22 and the total number of the population was 10,801. The total number of urine samples were analysed for iodine and thiocyanate were 1,032 (about 10%). The total goiter rate was 21.63%. Population of most of the studied areas had no biochemical iodine deficiency as evidenced by median urinary iodine excretion levels. However, the per capita consumption of iodine of about 40% population was inadequate. A large number of cyanogenic plants (SCN precursors) are used as common vegetables. This study ensures that the existing goiter prevalence in the region could possibly due to non-uniform adequate iodine supply along with the thiocyanate load. PMID- 11407153 TI - Advanced biliary atresia: is portoenterostomy justified in all infants? AB - During 1989-98, of the 127 patients with biliary atresia, 23 were seen with advanced biliary atresia (ABA) presenting with (i) at more than 120 days of age (ii) established cirrhosis and (iii) features of portal hypertension. Sixteen of these underwent exploratory laparotomy, dissection at the porta hepatis and hepatico-portoenterostomy (HPE) (group I). The remaining 7 infants underwent portal dissection and sump drainage only (instead of HPE). The drainage if any was evaluated for any change in colour, volume and concentration in the post operative period (group II). Age and presentation were the same in both the groups. In group-II, HPE was considered only if the bile flow was noted after portal dissection. Bile flow was seen in 3/16 in group I and 0/7 in group II. The fall in serum bilirubin during the first seven post operative days was noted in 2/16 in group I and 0/7 in group II. No drain output was recorded in any of the group II infants. The incidence of complications and the duration of hospital stay was significantly higher in group I patients. The sump drainage as an alternative procedure to HPE not only served the purpose of evaluating the patients with ABA for the possible bile flow in the post operative period but also avoided the need for a major operative procedure like HPE. PMID- 11407154 TI - Primary IgA nephropathy in children: association of clinical and pathological findings with prognosis. AB - Primary IgA nephropathy is a disease characterized by recurrent macroscopic or microscopic hematuria and diffuse mesangial IgA deposition. Although IgA nephropathy had previously been suggested to have a benign prognosis, long term follow-up of the patients revealed that it might lead to chronic renal failure. In this study, the association of the initial clinical and laboratory findings with the renal histological changes was evaluated in 14 cases with primary IgA nephropathy who were at follow-up with a mean duration of 43.07 +/- 16.88 months. Finally the correlation between the clinicopathological findings and prognosis was investigated. In 92.8% of the patients, macroscopic hematuria was the presenting complaint. Proteinuria was detected in 42.9% of the cases mild proteinuria in 14.3% and moderate in 28.6%. Renal biopsy specimens, evaluated according to Churg-Sobin's classification, showed grade 1 changes in 35.7% cases, grade 2 in 35.7%, grade 3 in 14.3% and grade 4 in 14.3%. Both the patients with grade 4 histology had moderate proteinuria, and developed chronic renal failure requiring hemodialysis. Prognosis was found to be associated with the degree of proteinuria and the severity of the histopathological findings. PMID- 11407155 TI - Attire and appearance of pediatrician: parents'/guardians' opinion. AB - Professional appearance of a pediatrician is an important element affecting the perception of competence of the doctor among patients and their parents/guardians, thus influencing their compliance. As there is no study on this issue addressing the Indian scene, we requested responses to a questionnaire containing 15 questions from the parents/guardians of the children. The purpose of the study was to assess their opinion regarding attire and appearance of pediatricians. Five each of the questions were related to common aspects of appearance and attire of the pediatrician in general and that of the male and the female pediatrician in particular. A total of 210 parents/guardians responded to see the question listed in the questionnaire. Assertive responses to each question were analysed for any association with the following categories--Urban, Rural, Males, Females, Urban Males, Rural Males, Urban Females, Rural Females, age below 40 years and age 40 years or more. Majority of the total assertive respondents preferred a formal/traditional attire and appearance for the pediatrician. Although most of the above categories concurred with this general opinion, some categories showed statistically significant, but practically unimportant, differences in the extent of agreement. The respondents generally favoured a white coat and nameplate on the pediatrician, shoes and a necktie for the male pediatrician and saree for the female pediatrician. They did not approve clothes with bizarre designs, use of perfumes by the pediatrician, long hair, T shirts and jeans pant for the male pediatrician, chudidar, lipstick and other make-ups, costly jewelery and short hair for the female pediatrician. PMID- 11407157 TI - Neurodevelopmental disorders and the pediatrician. PMID- 11407156 TI - Neuronal protection with magnesium. AB - Delayed neuronal death following hypoxic ischaemic insult is primarily mediated by the N-methyl D-aspartate (NMDA) receptor. The NMDA receptor antagonist MK 801, has been shown to limit neuronal death following hypoxic ischaemic injury but is too toxic to be used in the human neonate. Magnesium blocks the NMDA channel in a voltage dependent manner. Its administration after a simulated hypoxic ischaemic insult limits neurological damage in several animal models. The efficacy of magnesium in providing neuroprotection in the human neonate, however needs to be established in controlled clinical trials. PMID- 11407158 TI - Developmental disorders of activity dependent neuronal plasticity. AB - A number of neurodevelopmental disorders are caused by defects in activity dependent neuronal plasticity, the process by which neuronal activity shapes developing neuronal circuits. These disorders are caused by genetic mutations or other factors that disrupt intracellular signaling pathways that link the cell surface with the nuclear machinery for gene expression. The signaling pathways disrupted by these disorders are involved in learning, memory and behavior as well as in the synaptic proliferation and pruning that occurs during normal development. Examples of neurodevelopmental disorders that target plasticity include X-linked disorders such as Rett, Fragile-X and Coffin-Lowry Syndromes as well as acquired disorders such as cretinism. Several other X-linked mental retardation syndromes as well as autosomal disorders including neurofibromatosis type 1 and tuberous sclerosis also involve signaling pathways involved in neuronal plasticity. Disruption of neuronal plasticity is a mechanism that may underlie a diverse group of neurodevelopmental disorders. PMID- 11407159 TI - Pharmacotherapy of target symptoms in autistic spectrum disorders. AB - There are no aetiologically-based treatments available to cure autism. Though psychotropics have a role in the management of some symptoms of autism, clinical trial evidence for the use of psychotropics is in its infancy and needs close monitoring. About half of the subjects with high functioning pervasive developmental disorders (PDDs) are currently reported to be on psychotropics (anti-depressants, stimulants and antipsychotics), with many of them being on anti-epileptic medication simultaneously. Despite this high level of psychotropic use, few studies exist investigating the pharmacokinetics, pharmacodynamics or side-effect profiles in this population. Multiprofessional and parent partnership is essential in managing autism and psychopharmacology should be used in conjunction with environmental manipulation, educational modification and/or behavioral management strategies. A symptomatic approach to managing the difficult behaviours associated with autism is recommended. Some symptoms of autism may be medication responsive (hyperactivity, obsessions, rituals, inattention, tics, etc), while other symptoms may be responsive to behavioural interventions, but may require medication (aggression, anxiety, depression, impulsivity, sleep difficulties, etc), and symptoms which need specific skill remediation are usually non-responsive to medication (deficits in academic, social or sport domains). The new atypical antipsychotics (such as risperidone, olanzapine, amisulpiride, quetiapine) and SSRIs are increasingly being used in autism, with encouraging results, but a risk-benefit ratio of pharmacotherapy is essential with due weight being given to the side-effects of medication. Despite symptomatic improvement with medication, one should remain cautious about long term use of psychotropics. It is also important to recognize that psychotropics can sometimes worsen behaviour, and can produce iatrogenic symptoms. Certain anti epileptic medication and psychotropic drugs are metabolized by the same cytochrome P450 isoenzymes in the liver. In such circumstances, the addition of a psychotropic agent may drastically alter the levels of the anti-epileptic medication and vice versa. It is suggested that specialist clinics should be involved when one is considering complex medication regimes, experimental drugs, polypharmacy, or if patients show unusual side-effects or is drug resistant. PMID- 11407160 TI - Trends in cerebral palsy. AB - The terms trend and cerebral palsy are defined emphazing the non-diagnostic nature of the cerebral palsy label. Criteria necessary for valid estimation of trends include constant methods of estimating population based numerators and denominators over a number of years, which render them hard to obtain, particularly in developing countries. Trends in cerebral palsy are an important source of aetiological hypotheses for congenital cerebral palsy, provide corroborative evidence for existing hypotheses and may direct strategies to prevent post neonatally acquired cerebral palsy. In developed countries the overall frequency of congenital cerebral palsy has changed little during the last decades. However this masks a dramatic increase in the frequency in the infants born most preterm, a decline in those born moderately preterm and little change in those born at term, but the severity of impairments of those born very preterm is decreasing while for those born at term severity in increasing. These changes may be the result of the increasing ability of perinatal care to rescue very vulnerable infants. There is less agreement in trends of post neonatally acquired cerebral palsy which are more sensitive to social well being. PMID- 11407161 TI - Clinical considerations in the diagnosis of autism spectrum disorders. AB - The diagnosis of autistic spectrum disorders (ASD) is being made more frequently in children and at younger ages. This paper discusses various factors to be considered in the screening of autism, early features of presentation, relevant to assessment and diagnosis, subtypes or different syndromes within the spectrum of autistic conditions including Asperger syndrome, the differential diagnosis from learning and language disorders and the medical and behavioural commonly associated disorders. PMID- 11407162 TI - Congenital myotonic dystrophy. AB - Myotonic muscular dystrophy is the most frequent autosomal muscular dystrophy affecting adults and children. It affects multiple organ systems and is probably the best example of variable expressivity in a human disease. This article presents a patient with congenital myotonic dystrophy who had facial dysmorphism, hypotonia, talipes, feeding and respiratory difficulties in the neonatal period and later presented to us with developmental delay and had percussion myotonia. His mother had clinical and electrophysiological features of myotonia. Expansion of unstable CTG trinucleotide repeat in the myotonic protein kinase gene was demonstrated in both. The identification of this molecular defect allows its specific diagnosis in relation to other neuromuscular disorders as well as accurate prenatal diagnosis. PMID- 11407163 TI - Mobius syndrome associated with ventricular septal defect. AB - We report a 10-year-old boy with Mobius syndrome (MS) associated with ventricular septal defect who was delivered after an unsuccessful curettage before the 10th week of gestation. Methylergobasine which is an ergot alkaloid was also applied during the curettage procedure. Despite of the curettage procedure, the pregnancy was continued and he was delivered. His developmental milestones were delayed. On account of this case, we think that direct mechanical fetal trauma and vasoconstriction or both may cause MS. But, further extensive studies are needed to verify this hypothesis. PMID- 11407164 TI - Leucocytoclastic vasculitis in infant. AB - Acute hemorrhagic edema of infancy is a laucocytoclastic vasculitis characterized by a triad of fever, large purpuric skin lesions and edema. The presentation is often dramatic. The manifestations are often misinterpreted as a septicemic illness. We report the occurrence of this syndrome in a 1 1/2-year-old-child, highlighting some of the diagnostic and therapeutic problems encountered. PMID- 11407165 TI - Infantile hemangioendothelioma. AB - Primary hepatic tumors are uncommon in children and account for only three per cent of the tumors in children. Infantile hemangioendothelioma is a rare benign hepatic tumor arising from mesenchymal tissue. Most of the cases present before six months. An unusual presentation and progression of infantile hemangioendothelioma is reported in a 19-month-old female child. The diagnosis was arrived at by radiological and histopathological examination. The patient underwent excision surgery, following which made an uneventful recovery. On follow-up at six months, patient was asymptomatic with no evidence of recurrence. PMID- 11407166 TI - Ptosis in late infantile Tay-Sachs disease. AB - The brief communication describes a 2-year-old child who presented with delayed achievement and regression of milestones, seizures of multiple types, exaggerated response to sound, inability to see and bilateral cherry red spots. In addition to these typical manifestations of the late infantile variety of Tay-sachs disease, unilateral ptosis was present. The magnetic resonance imaging of brain revealed abnormalities consistent with an advanced stage of the disease. PMID- 11407167 TI - Proboscis lateralis of nose. AB - A rare case of congenital malformation of the nose that was successfully corrected surgically is reported. PMID- 11407168 TI - Iodine deficiency disorders. PMID- 11407169 TI - Social capital, disorganized communities, and the third way: understanding the retreat from structural inequalities in epidemiology and public health. AB - The construct of social capital has recently captured the interest of researchers in social epidemiology and public health. The authors review current hypotheses on the social capital and health link, and examine the empirical evidence and its implications for health policy. The construct of social capital employed in the public health literature lacks depth compared with its uses in social science. It presents itself as an alternative to materialist structural inequalities (class, gender, and race) and invokes a romanticized view of communities without social conflict that favors an idealist psychology over a psychology connected to material resources and social structure. The evidence on social capital as a determinant of better health is scant or ambiguous. Even if confirmed, such hypotheses call for attention to social determinants beyond the proximal realm of individualized sociopsychological infrastructure. Social capital is used in public health as an alternative to both state-centered economic redistribution and party politics, and represents a potential privatization of both economics and politics. Such uses of social capital mirror recent "third way" policies in Germany, the United Kingdom, and United States. If third way policies lose support in Europe, the prominence of social capital there might be short lived. In the United States, where the working class is less likely to influence social policy, interest in social capital could be longer lived or could drift into academic limbo like other psychosocial constructs once heralded as the next big idea. PMID- 11407170 TI - Defusing the baby boomer time bomb: projections of after-tax income in the twenty first century. AB - There has been a major national policy debate over the prospective tax burden facing future generations of workers as a result of the impending retirement of the baby boom generation. However, the real determinant of well-being is after tax income, not the tax rate on before-tax income. This study constructs a series of projections of after-tax income, for workers and families at different points in the income distribution, to examine the effects of several different trends. The author first calculates the extent to which after-tax income can be expected to decline as the result of the aging of the baby boomers, then calculates the impact on after-tax income for families at different points in the income distribution of a continuation of recent trends in wage inequality. He also constructs a category of "after-tax, after-health-care spending" income, which examines the impact of the continued rapid growth in health care costs. The findings suggest that the continuation of recent trends in inequality and health care cost growth will have a much larger effect on future living standards than will the aging of the baby boomers. PMID- 11407171 TI - Health care workers' unions and health insurance: the 1199 story. AB - Labor unions that represent health care workers encounter unique circumstances. This study focuses on 1199--the largest health care workers' union in the United States, based in New York City--and describes how the union and its National Benefit Fund have structured a managed indemnity health insurance program that provides extensive benefits for its members. The authors detail the workings of the National Benefit Fund and the ways in which it can improve the health care of its members through its union structure. The Fund represents a model for improving the provision of health care to Americans. PMID- 11407172 TI - Health reform in Mexico: the promotion of inequality. AB - The Mexican health reform can be understood only in the context of neoliberal structural adjustment, and it reveals some of the basic characteristics of similar reforms in the Latin American region. The strategy to transform the predominantly public health care system into a market-driven system has been a complex process with a hidden agenda to avoid political resistance. The compulsory social security system is the key sector in opening health care to private insurance companies, health maintenance organizations, and hospital enterprises mainly from abroad. Despite the government's commitment to universal coverage, equity, efficiency, and quality, the empirical data analyzed in this article do not confirm compliance with these objectives. Although an alternative health policy that gradually grants the constitutional right to health would be feasible, the new democratically elected government will continue the previous regressive health reform. PMID- 11407173 TI - Economic growth, income equality, and population health among the Asian Tigers. AB - The "Tiger" economies of Southeast Asia provide examples of developing nations where economic growth and increasing income equality are compatible and, when occurring together, are associated with superior health trends over time. The degree of income inequality in the Asian Tigers declined during the period of rapid economic growth. Traditionally, economists have viewed economic growth and relative parity in income distribution as incompatible, or trade-offs. This poses a public policy dilemma, since a reasonable propensity to increase a nation's overall economic well-being would mean forsaking measures that increase income parity. The Asian Tigers, however, have shown that this need not be viewed as a trade-off. Economic growth and a simultaneous increase in income equality are possible and, with respect to health outcomes, desirable. The authors propose a variety of mechanisms through which income inequality can enhance economic growth, and discuss policies in education, agricultural land reform, and housing that influence the simultaneous attainment of income equality and economic growth. PMID- 11407174 TI - The global expansion of precarious employment, work disorganization, and consequences for occupational health: a review of recent research. AB - In this review of a range of studies on the health and safety effects of precarious employment in industrialized societies published since 1984, the authors examine the overall findings and methodological issues and identify areas in need of further research. Of the 93 published journal articles and monographs/book chapters reviewed, 76 studies found precarious employment was associated with a deterioration in occupational health and safety (OHS) in terms of injury rates, disease risk, hazard exposures, or worker (and manager) knowledge of OHS and regulatory responsibilities. Of the more than 25 studies each on outsourcing and organizational restructuring/downsizing, well over 90 percent find a negative association with OHS. The evidence is fairly persuasive for temporary workers, with 14 of 24 studies finding a negative association with OHS. The evidence is less strong for small business, and a handful of studies on part-time workers found no clear association with negative OHS outcomes (in some cases the reverse). Further research is needed to more clearly link health effects to particular business practices and neoliberal policies and to explore the regulatory implications of the growth of precarious employment. The authors suggest some ways to conceptualize the association between precarious employment and occupational health. PMID- 11407175 TI - "The worst of both worlds": the management reform of the World Health Organization. AB - The governance and management of global health institutions, such as the World Health Organization (WHO), are under increasing critical scrutiny. This management case study explores the first year of transformation at the WHO under Director-General Dr. Gro Harlem Brundtland, focusing on the key stakeholders and the role of complexity in institutional change. This is a story about transition in a difficult, politically fraught, and management-resource-constrained environment. In the search for appropriate management paradigms, organizations such as the WHO may believe that the answers lie in harsh reengineering and the search for high-profile "success stories." Ironically, global business has moved away from such approaches and is far more focused on collaboration, empowerment, and knowledge sharing. PMID- 11407176 TI - International Monetary Fund and World Bank policies can hurt poor, study says. AB - According to a study by a World Bank economist, developing countries that do not follow International Monetary Fund and World Bank economic programs have seen more of their people lifted out of poverty in times of economic growth than have countries that do take the advice of these lenders. PMID- 11407177 TI - In Mozambique, a less than helpful hand. AB - World Bank loan policies in Mozambique have almost killed off the country's cashew-processing industry by requiring the removal of the export tariff on raw cashews, a tax that Mozambique had instituted to ensure the cashew supply for local factories. PMID- 11407178 TI - Putting the search for genes in perspective. AB - The sequencing of the human genome has been heralded by both the mass media and scientists as a breakthrough that will allow the detection of individuals at increased risk for common diseases and the tailoring of drugs to an individual's genetic profile in order to prevent disease. Sequencing is likely to benefit those at risk of developing rare diseases in which inherited mutations in a single gene play a major causal role. In the vast majority of people with common diseases, however, genotypes at many different loci, as well as environmental exposures, must be simultaneously present before disease appears. Elucidating the genes involved will prove elusive. In addition to the large number, different combinations account for a particular disease. Most of the genotypes that contribute to the constellation of necessary genes are uncommon and will be difficult to find. Common genotypes may confer susceptibility but will be weak predictors of disease. Because of the difficulty of discovering genes for common diseases, designing therapies will also prove difficult. More attention to environmental risk factors for particular diseases will have greater yield than a genetic search, but this too will be difficult because of environmental-genetic and other interactions. The search for risk factors for particular diseases neglects the political and social milieu in which individuals swim or sink and in which all diseases occur. PMID- 11407179 TI - Family social support and prenatal care among unmarried African American teenage primiparas. AB - This study determined the relation of family social support and the receipt of adequate prenatal care among 25 unmarried African American teenage primiparas residing in the home. The study also investigated the strength of the family social support system compared to nonfamily support systems. Using the Personal Resource Questionnaire, Part II to measure social support, participants were provided the questionnaire 1 to 2 days after birth, but immediately prior to leaving the hospital. Results of the study determined no significant difference in social support scores among those women who received either adequate or inadequate prenatal care, although the 17- to 18-year-old age group had higher support scores than the 15- to 16-year-old age group. However, findings from the study indicated that family social support between both groups was significantly greater than nonfamily support systems. Thus, interventions by community health nurses to involve family members in all aspects of birthing and parenting can be essential in promoting positive health outcomes. PMID- 11407180 TI - A smoking reduction and cessation program with registered nurses: findings and implications for community health nursing. AB - A smoking reduction and cessation program was implemented with registered nurses in 3 Canadian provinces. Nurses (n = 117) participated in either an 8-week group or self-directed program using a resource specifically designed for nurses. Questionnaires were administered prior to and at the end of the 8-week interventions and at 6 and 12 months postintervention. Statistically significant changes at 8 weeks in nurses' smoking practices were found on the number of nurses continuing to smoke, mean number of cigarettes smoked, and movement in the stage of behavioral change. Attrition and variation in patterns of quitting over the 12-month study period made assessing participants' longer term outcomes difficult. This study highlights the complexity of assisting nurses to quit smoking and of implementing and evaluating a program based on accepted community health models of practice. PMID- 11407181 TI - Prevalence of diabetes mellitus and related conditions in a south Texas Mexican American sample. AB - The prevalence of self-reported diabetes mellitus in a Mexican American sample population living in Hidalgo County in south Texas was examined along with risk for related sequelae. Data from a stratified random sample of 849 Mexican American men and women ages 45 and older were examined. A subsample of 193 diabetics was compared to 656 nondiabetics with respect to hospitalization, reasons for hospitalization, and other related medical conditions. The prevalence of self-reported diabetes for both men and women was 25.9%. Hospitalization rates for diabetics over a 1-year period and a 5-year period were significantly higher than nondiabetics. Hypertension, cardiac problems, amputations, kidney or urinary problems, eye problems, and joint and bone problems were significantly more common in the diabetics sample than in the nondiabetic sample. The data indicate that diabetes in Mexican Americans may be undertreated in south Texas. PMID- 11407182 TI - Undergraduate community health nursing education in a neighborhood settlement house. AB - Undergraduate community health nursing faculty are continually challenged to find appropriate clinical placements. Prior to using an inner-city neighborhood settlement house as a clinical site, traditional public health and home-care agencies provided clinical placements for senior baccalaureate nursing students. The settlement house had several programs but did not have a traditional nursing department. A 14-item questionnaire was developed to examine student (N = 30) perceptions of the opportunities for course objectives to be met and student attitudes and beliefs regarding the settlement-house placement. The majority of the students agreed that the learning experiences at the settlement house provided opportunities to meet course objectives. Findings indicated that students who considered the neighborhood safe are more apt to recommend the clinical placement to other students (p < .01). Those students who believe that professional nursing services can help improve the health of people who live in the inner city are more apt to say, "This experience changed my beliefs regarding people who live in an inner-city neighborhood" (p < .01). PMID- 11407183 TI - Healthy eating for rural low-income toddlers: caregivers' perceptions. AB - Caregivers exert a powerful influence on young children's eating habits. This qualitative study used focus groups to assess nutritional needs and barriers in establishing healthy eating habits in toddlers. Three focus groups were conducted with rural, low-income caregivers, 2 with men and 1 with women, in 3 rural Michigan counties. Four major themes emerged: (a) barriers to providing healthy meals, (b) division of responsibility, (c) mealtime behavior, and (d) desired nutrition education. The major barriers identified were work schedules; cost of food; inadequate time to shop, plan, and prepare nutritious meals; or a combination thereof. Caregivers expressed concern for the nutritional well-being of their toddlers. The perceived needs and perceptions of low-income caregivers need to be considered when providing nutrition education. Findings from this study provided the basis for developing a nutrition education intervention for low-income parents of young children. PMID- 11407184 TI - New Zealand pathologists: a case study in occupational control. AB - This paper examines the progressive exertion of external managerial control over New Zealand pathologists as the country's New Public Management health reforms were implemented during the 1990s. Perspectives on professionalism, and its role in the effective use of resources, are discussed as part of the examination of this shift in decision-making power from pathologists to external management. Our analysis, based on a range of archived and interview data collected over the period 1997-2000, suggests that publicly unacceptable compromises in pathology service quality were risked by the pursuit of tight bureaucratic and free market controls over pathology practice. The paper concludes with suggestions for a health professional control model facilitative of maximal health gain. PMID- 11407185 TI - Performance implications of Porter's generic strategies in Slovak hospitals. AB - The aim of the study was to examine the use of Porter's generic strategies and their effect on performance in the context of the Slovak hospital industry. Using mail survey the study first identified the natural taxonomy of four strategic types of Slovak hospitals, based on their use of Porter's generic strategies in pure form and in combination. Next the study examined whether different strategic types were associated with different levels of organisational performance, while controlling for such variables as size and location, which have been argued to influence the hospital performance. The findings indicate that hospitals which follow a "stuck-in-the-middle" strategy, in general, have superior performance on all used performance measures, while hospitals that place only low emphasis on cost leadership, differentiation and focus, labelled "wait and see" in this study, perform the poorest. The study concludes that the research provided body of knowledge relevant for the Slovak hospital industry, that may be used by hospital managers in the strategy formulation process as well as by the researches in exploring the influence of different contingencies on hospitals' strategic orientation. PMID- 11407186 TI - New roles and relationships in the NHS--barriers to change. AB - The incoming Labour Government's vision for reforming the NHS in Scotland was outlined in the White Paper Designed to Care. While bearing similarities to the proposals outlined for the rest of the UK, it also had distinctive differences. Organisational structures, roles, and relationships between the different parts of the NHS were to be fundamentally altered, particularly in primary and community care. This paper reports upon a series of interviews undertaken across several Health Board areas, with key stakeholders involved in the primary and community sectors. These interviews were intended to examine the development and evolution of the new organisational arrangements, and to identify potential barriers to the successful implementation of Designed to Care. Several barriers and sources of institutional resistance to the new roles and relationships were found during this study, and are discussed. Suggestions upon how these may be overcome and implementation improved are then made. PMID- 11407187 TI - Effective leadership behaviour: leading "the third way" from a primary care group perspective. A study of leadership constructs elicited from members of primary care group management boards. AB - The UK National Health Service (NHS) is undergoing cataclysmic change following the election of the first Labour Government in 18 years. This is primarily embodied in the implementation of the White Paper The New NHS Modern-Dependable, which has resulted in the creation of primary care groups (PCGs) and primary care trusts (PCTs). The task facing both PCGs and PCTs is a radically new and complex one, requiring a new set of leadership skills to the traditional command and control style management. Leadership theories have evolved over the past 70 years. However, it was not until the 1980s that a major change in the paradigm of thinking around what is the nature of leadership occurred. The interaction between the leader and his/her followers is explored in what has become known as transformational leadership theories, developed by Bass and Avolio. Recent studies have, however, questioned the applicability of leadership models derived in the USA, to other cultures. This paper explores the leadership behaviours required for the management boards of PCGs and PCTs. A qualitative research method "Grounded Theory" approach was chosen for this study of leadership. The Repertory Grid technique was used to collect data. There are a number of implications arising from the findings of this study for both leadership models in general, and more specifically, for the development of leadership skills in both PCGs and PCTs. PMID- 11407188 TI - Death plans. PMID- 11407189 TI - Attitudes and knowledge about cancer pain in Flanders. The educational effect of workshops regarding pain and symptom control. AB - Despite international agreements and recommendations regarding cancer pain therapy, the effectiveness of pain treatment is still a major problem even in Western countries. Part of the problem is that physicians and nurses often lack knowledge of methods for the assessment and treatment of cancer pain and may have many rigid beliefs and attitudes. This study investigated the misconceptions of physicians and nurses that play a role in the undertreatment of pain in Flanders (Belgium). We approached 197 health care workers who participated in the pain and symptom control education sessions organized by the Federation Palliative Care Flanders, and asked them to complete a questionnaire both before and after the sessions. The impact of the education sessions on their knowledge and beliefs regarding the management of cancer pain was substantial. Methods of reaching the target groups that do not feel the need for further education are discussed. PMID- 11407190 TI - Educational opportunities in palliative care: what do general practitioners want? AB - It is important to support general practitioners (GPs) in maintaining and developing their palliative care skills as most of the final year of a patient's life is spent at home under the care of the primary health care team. The training needs and uptake of GPs have been explored, but little is known about how GP educational preferences vary. The aim of this study was to explore the current educational preferences of GPs in different geographical locations as part of an evaluation of an educational intervention. The methods used included postal questionnaires sent to 1061 GPs. Results from 640 (60%) of GPs revealed that half (51%) wanted education in symptom control for non-cancer patients. More inner-city GPs wanted education in opiate prescribing (43%), controlling nausea and vomiting (45%), and using a syringe driver (38%) than their urban and rural colleagues (26%, 29% and 21%, respectively). Increased educational preference and increased difficulty in accessing information was associated with reduced confidence in symptom control. To maximize educational uptake it will be important for educational strategies to be developed and targeted according to variations in demand, and in particular to respond to the need for palliative care education in symptom control for patients suffering from advanced non malignant disease. PMID- 11407191 TI - Communication Capacity Scale and Agitation Distress Scale to measure the severity of delirium in terminally ill cancer patients: a validation study. AB - Although valid measurement of the severity of terminal delirium is of great importance in palliative care settings, existing instruments have considerable limitations. In order to quantify patients' communication capacity and agitated behaviour, two new operational observer-rating scales, the Communication Capacity Scale (Communication Scale) and Agitation Distress Scale (Agitation Scale), were validated. Thirty terminally ill cancer patients diagnosed with delirium were evaluated simultaneously by two palliative care physicians blinded to each other's coding using the Communication Scale and Agitation Scale. In addition, the Memorial Delirium Assessment Scale (MDAS), Delirium Rating Scale (DRS) and Sedation Scale were rated by one researcher. Both scales achieved high internal consistency and inter-rater reliability with Cronbach's alpha coefficients of 0.91 and 0.96, and Cohen's kappa values on each item of 0.72-1.00. The principal components analysis resulted in the emergence of only one component for each scale. The total score on the Communication Scale was highly associated with that of the MDAS (rho = 0.78), Sedation Scale (rho = 0.86), and cognitive items from the MDAS and DRS (rho = 0.83). The whole score on the Agitation Scale was significantly correlated with that of the DRS (rho = 0.61) and agitation items from the MDAS and DRS (rho = 0.61). In conclusion, the Communication Scale and Agitation Scale have acceptable reliability and validity to quantify patients' communication capacity and agitation symptoms of terminally ill cancer patients with delirium. PMID- 11407192 TI - Mainstreaming palliative care for cancer patients in the acute hospital setting. AB - Palliative care is now emerging as an integrated part of mainstream health care delivery. The importance of patient choice regarding place of dying means that a substantial proportion of palliative care provision occurs in community settings. In part, this is due to the inappropriateness of the acute hospital setting for the care of dying patients. However, most patients with cancer and other terminal illnesses are diagnosed and treated in acute hospitals. Acute hospitals are also the most common setting where people actually die. Therefore, there remains a need for skilled and compassionate provision for the care of dying patients in the acute hospital setting. This paper presents a case for the provision of palliative care services in teaching hospitals. It further argues that a high level of integration between cancer treatment services and palliative care services is needed to optimize the care of cancer patients. PMID- 11407193 TI - Lung cancer health care needs assessment: patients' and informal carers' responses to a national mail questionnaire survey. AB - The objective of this study was to describe patients' and informal carers' perceptions of care received and services offered following a diagnosis of primary lung cancer. We prepared a prospective, national, mail questionnaire survey of 466 patients with a diagnosis of primary lung cancer and a lay carer of their choice. The setting was 24 randomly chosen hospitals throughout the UK, from a range of urban (n = 11) and rural settings (n = 13). The majority (76%/159) of responders were recipients of care from cancer units. Two hundred and nine patients (45%) with primary lung cancer and 70 (15%) lay carers completed questionnaires. The main results that we found were that key areas of unmet need were most apparent during periods away from acute service sectors, with as few as 40% of patients reporting having received as much help as they needed from community services. The greatest onus of care for patients fell to lay carers, but only 29% of patients identified their lay carers as having needs in relation to their illness. Where patients received all their diagnostic tests in one hospital they were significantly more likely to wait less time between first seeing their general practitioner (GP) and being told their diagnosis (P = 0.0001) than patients who had to attend more than one hospital during their diagnostic work-up period. Fifty per cent of patients reported experiencing some degree of breathlessness even at rest, but only 15% reported having received any advice on living with it. Less than a quarter (23%) of hospital consultants identified anxiety as a key problem for patients with lung cancer, but 66% of patients identified it as such. Hospital staff largely overlook the needs of informal carers, who derive support from a small, mainly community oriented group of professionals, but accessing help is problematic and is dependent on local resources and a need to be proactive. Our conclusions are that developments in service provision for patients with lung cancer and their informal carers need to focus on six key areas: development of strategies to encourage patients to present earlier to their GP; ongoing evaluation of rapid diagnostic clinics; development and evaluation of a lung cancer care coordinator role; evaluation of innovations in delivery of nursing care in the community; development of local guidelines to facilitate equitable access to palliative care and social services; and evaluation of supportive strategies targeted at lay carers. PMID- 11407194 TI - Chaplaincy in hospice and hospital: findings from a survey in England and Wales. AB - The purpose of this study was to identify and compare key features of spiritual care provision in hospices and acute hospitals within England and Wales. A survey by postal questionnaire was carried out on the perceptions of senior chaplains in 151 hospices and 194 hospital trusts. The overall response rate was 76%. The findings of the survey suggest (a) a trend away from establishing chapels as the spiritual focal point within hospitals and hospices, (b) the development of broader roles for chaplains, (c) a different level of service provision between hospices with funded chaplaincies and hospices with voluntary chaplaincies, (d) a wide range of non-religious spiritual care requirements on the part of patients and (e) a more frequent requirement for religious care in hospitals rather than hospices. PMID- 11407195 TI - Prevalence and characteristics of breakthrough pain in patients with non malignant terminal disease admitted to a hospice. AB - A prospective survey was undertaken to determine the prevalence and characteristics of breakthrough pain in patients with non-malignant terminal disease admitted to a hospice. Of the 78 admissions surveyed, 10 patients were confused or too unwell to take part and 25 were pain-free. The remaining 43 reported 86 pains (range 1-6 per patient); of these patients, 27 (63%) had breakthrough pain and identified 52 pains (range 1-5 per patient). Breakthrough pain was classified as somatic (46%) visceral (14%), neuropathic (25%) or mixed aetiology (15%); 60% of pains were severe or excruciating. The mean number of daily breakthrough pain episodes was five (range 1-13), 54% of which occurred suddenly. Most pains (56%) were unpredictable; 75% lasted less than 30 min. These findings suggest that breakthrough pain is common in patients with non-malignant terminal disease; it is frequent, short lasting and often unpredictable, thus making treatment difficult. PMID- 11407196 TI - Use of clinical pictures in the management of nausea and vomiting: a prospective audit. AB - This prospective audit aimed to evaluate a guideline for the management of nausea and vomiting in palliative care inpatients. Clinical pictures were used to guide diagnosis and treatment, with potentially reversible causes being addressed where appropriate. Over a 3-month period, 40 patient episodes occurred, all of which were included in the audit. The commonest clinical pictures were gastric stasis/outlet obstruction (35%) and chemical/metabolic (30%). Management according to the guideline was effective. Nausea was abolished in 28 of 34 cases (82%) and vomiting resolved completely in 26 of 31 cases (84%). Symptoms were totally controlled in a mean time of 3.4 days. Nausea and vomiting, although distressing symptoms, can be controlled in the majority of cases. Multi-centre prospective audit, using a standardized tool, may prove useful in allowing larger numbers of patients to be systematically analysed and individual centres to compare outcomes. PMID- 11407197 TI - Risperidone: a useful adjunct for behavioural disturbance in primary cerebral tumours. PMID- 11407198 TI - Assessing hospice inpatients with pain using numerical rating scales. PMID- 11407199 TI - The formation of ECEPT (Eastern and Central Europe Palliative Task Force) a Polish initiative. PMID- 11407200 TI - Clinical research in palliative care: choice of trial design. AB - Prospective controlled clinical trials provide the highest level of evidence in palliative care. Both parallel and crossover studies have unique advantages and disadvantages. Excellent knowledge of the patient population, interventions and endpoints are useful in choosing the most appropriate trial. Open pilot studies can assist the investigator in understanding the intervention and measurement tools better before embarking in costly and lengthy controlled trials. More controlled clinical trails are need to generate the evidence that will advance patient management in palliative care. PMID- 11407201 TI - Reliability of a questionnaire for evaluating the understanding of asthma. AB - The present study was designed to evaluate the repeatability of a questionnaire developed to assess the understanding that asthma patients have of their disease and, on the basis of its variability, estimate the sample size necessary for determining the efficacy of a future structured program on asthma knowledge. The repeatability of the Asthma Questionnaire (AQ) was evaluated by asking 89 patients to complete it twice within a period of 7-10 days without the subject being exposed to any programme on asthma knowledge between the two administrations. The AQ was demonstrated to have good content and face validity. Results showed that neither age nor sex had a significant influence on total scores, and that the degree of reliability was adequate (R = 0.769). The mean percentage of correct answers was observed to be approximately 70% in both sessions, suggesting a consistent area for possible improvement which could be targeted by means of an appropriately structured programme on asthma knowledge. For comparative purposes before and after the programme, or for measuring its efficacy, the AQ should be recommended. In conclusion the Asthma Questionnaire could provide a useful tool for the general practitioner, chest physician and other health professionals, to assess what the patient really does understand or does not, concerning asthma management, and hence be the starting point for a well-tailored educational intervention. PMID- 11407202 TI - Quality of Life Evaluation and Survival Study: a 3-yr prospective multinational study on patients with chronic respiratory failure. AB - Therapy of patients with chronic respiratory failure is mainly directed at minimizing symptoms in order to improve, or at least to prevent a deterioration of, patients' well-being. Under such circumstances, the perceived effect of therapies on patients' well-being and daily life represents the most important subjective outcome of treatment. Therefore, there is a need to provide a global estimate of health in patients on long term oxygen therapy or overnight home mechanical ventilation. The Maugeri Foundation Respiratory Failure Questionnaire (MRF28) is the first health status ("quality of life") questionnaire specifically developed for use in CRF and its items were selected to be applicable to patients with both obstructive and restrictive diseases. The Quality of Life Evaluation and Survival Study (QuESS) is a multinational study with the aim of re-evaluating the natural history of chronic respiratory failure in about 300 patients. To the authors knowledge, the Quality of Life Evaluation and Survival Study is the first study to evaluate the natural history of chronic respiratory failure in such a large number of subjects and with a complete set of data. In fact, both pathophysiologic and health status assessments will be made. Moreover, by collecting data on mortality, disease exacerbations and hospitalization, it will also be possible to verify the predictive ability of health status versus pathophysiology in terms of mortality and healthcare utilization. PMID- 11407203 TI - Dyspnoea exaggerated in the supine position and during exertion--diagnostic challenge. AB - The case of dyspnoea, exaggerated when in the supine position and during exertion, as a result of severe weakness of the diaphragm is reported. The aim of the study was to present a rare case of idiopathic bilateral diaphragmatic paresis (BDP) and to describe all the diagnostic procedures necessary to perform differential diagnostics. In order to establish the final diagnosis, chest radiography, haemodynamic evaluation of the circulatory system, ultrasonography, ultrasonocardiography, measurement of transdiaphragmatic pressures, scintiscanning of the lungs, spirometry, analysis of arterial blood gases, computed tomography of the thorax and external stimulation of the phrenic nerve were performed. The measurement of transdiaphragmatic pressure was crucial to establish and confirm the diagnosis of BDP, as only a small difference in gastric and oesophageal pressures during tidal breathing and inspiratory efforts was recorded. As no cause of diaphragmatic paresis was found, the case was classified as idiopathic. The final diagnosis of non-trauma related bilateral diaphragmatic weakness was generally delayed. In the case of the described patient, dyspnoea, the main symptom he was suffering from, was supposed to result from his congenital heart defect. We recommend that the suspicion of idiopathic diaphragmatic paresis should always be raised in patients suffering from respiratory failure of unknown origin. It is, however, necessary to perform extensive diagnostics to exclude the other causes of phrenic-diaphragmatic impairment. It's also necessary to consider all infections, injuries and surgical procedures within the thorax as possible causes of diaphragmatic paresis. PMID- 11407204 TI - Successful treatment of endobronchial amyloidosis using Nd:YAG laser therapy as an alternative to lobectomy. AB - Endobronchial amyloidosis may cause local obstruction associated with a variety of clinical presentations including recurrent respiratory infection. Although some patients may require surgical intervention, encouraging experience with endobronchial Neodymium (Nd):YAG laser therapy in the management of this condition is reported. Two patients with localized endobronchial amyloidosis were treated with endobronchial Nd:YAG laser therapy. One patient received two treatments for right upper lobe amyloid and another patient received three treatments for right lower lobe amyloid. Excellent clearance of amyloid was achieved in each patient. The first patient remains well, 16 months post treatment with full patency of the apical and posterior segments. Ten months post treatment, the second patient has full patency of the right lower lobe. No complications occurred in association with Nd:YAG laser therapy. Endobronchial Neodymium:YAG laser treatment should be considered for selected patients with localized endobronchial obstruction due to amyloidosis. PMID- 11407205 TI - A molecule across centuries. PMID- 11407206 TI - Tuberculosis and lung cancer. An interesting case study. AB - This report describes the case of a patient with lung cancer who completely recovered when he was suffering from tuberculosis. Since bacillus Calmette-Guerin (BCG) has beneficial effects in certain types of cancer, it was hypothesized that infection with Mycobacterium tuberculosis induced an effective response against the tumour. M. tuberculosis-infected blood T-lymphocytes of the patient were cultured with two lung tumour cell lines. T-lymphocytes in vitro remained attached to tumour cells that appeared reduced in number. Moreover, M. tuberculosis isolated from the patient was a strong inducer, in infected macrophages, of the expression of the inducible form of the nitric oxide synthase, that may regulate cytotoxic activity of human macrophages. PMID- 11407207 TI - What is evidence-based medicine and the role of the systematic review: the revolution coming your way. AB - The practice of evidence-based medicine involves the critical application of current best evidence to the care of individual patients. This process requires clinicians to be able to efficiently locate, critically appraise and appropriately apply the best available evidence to particular clinical scenarios. The current volume of medical and scientific literature is unmanageable and traditional approaches to continuing education cannot fully address the information needs of practicing clinicians in the era of evidence-based medicine. Review articles are useful resources but traditional narrative reviews often include a selective summary of research findings mixed with personal opinion. Systematic reviews have many advantages over traditional narrative reviews; importantly, the use of systematic and explicit methods mean that bias in identifying, selecting and summarizing the evidence is minimized. For systematic reviews where meta-analysis can be performed, the quantitative synthesis of data from different studies can provide additional information. Systematic reviews are retrospective studies and like all research may be susceptible to bias. Furthermore some methodological issues related to meta-analysis have yet to be resolved. The strengths and limitations of systematic reviews are discussed in this article and useful resources for locating high quality systematic reviews are outlined. Clinical judgement and experience have a pivotal role in the practice of evidence-based medicine and clinicians must rely on this judgement when deciding how and when to apply the results of systematic reviews to particular patients. PMID- 11407208 TI - Measurement of dyspnoea. AB - Dyspnoea is the term generally applied to unpleasant or uncomfortable respiratory sensations. The measurement of dyspnoea is a critical aspect of patient evaluation and management. Both psychophysical methods and clinical scales have been used to assess breathlessness. Because of dyspnoeic patient is frequently unable to perform the daily activities of life due to discomfort associated with breathing, the clinical methods used to measure dyspnoea have depended primarily on the magnitude of the exertional task that evokes breathlessness. The visual analogue scale is one method that has been used. To improve the clinical rating, two more indices (baseline dyspnoea index and transition dyspnoea index) that include the components of functional impairment and magnitude of effort, in addition to the magnitude of the task, were developed. The category-ratio-10 scale developed by Borg consists of verbal descriptors adjacent to specific numbers, providing a category scale with ratio properties. Measurement instruments may involve a cost for use, may be self-administered or require an interviewer, and will vary in the time required for completion and scoring. PMID- 11407209 TI - Pharmacological treatment of exercise dyspnoea. AB - A better understanding of the mechanisms of dyspnoea improves the clinician's ability to treat patients with shortness of breath. Any intervention that: 1) reduces ventilatory demands; 2) reduces ventilatory impedance; or 3) improves inspiratory muscle function, may relieve dyspnoea. Reduced ventilatory demand may be obtained by reducing metabolic load. Supplemental oxygen during exercise reduces exertional breathlessness and improves exercise tolerance, the decrease in dyspnoea being proportional to decrease in minute ventilation. Reduced ventilatory demand may also be obtained by decreasing the central drive. Opiates have been shown to decrease minute ventilation at rest and during submaximal exercise. They can alter the central processing of neural signals within the central nervous system to reduce sensations associated with breathing. Contrastingly, no consistent improvement in dyspnoea (versus placebo) has been shown with anxolytics. Decreasing central drive may also be obtained by altering pulmonary afferent information. Interventions that alter transmittal of afferent information to the central controller, potentially reduce dyspnoea. Reduction of ventilatory impedance is obtained by administering B2, anticholinergics or theophylline. B2 and anticholinergics act by modulating the increase in operational lung volumes and the inspiratory muscle effort during exercise. The mechanism by which theophylline relieves dyspnoea is probably related to a mechanism other than its bronchodilation alone. Alterations in respiratory muscle function are currently being detected in patients with chronic obstructive pulmonary disease, due to alteration in respiratory muscle energy balance. Nutritional repletion may improve respiratory muscle function but uncertainty remains as to whether nutritional repletion may relieve dyspnoea. The cumulative benefit of interventions targeting the pathophysiologic mechanism of dyspnoea must be identified for optimum treatment of patients with shortness of breath. PMID- 11407210 TI - The bronchial epithelium in asthma--much more than a passive barrier. AB - The bronchial epithelium has a multifunctional role in the airway. It is actively engaged in communicating with cells of the immune and inflammatory systems, as well as secreting cytoprotective molecules and acting as a physical barrier between the internal and external milieu of the lungs. In asthma, the bronchial epithelium is often damaged, with shedding of the columnar cells into the airway lumen. This damage and ensuing repair responses are proposed to orchestrate airway remodelling via activation of myofibroblasts in the underlying lamina reticularis. This allows the two cell types to work as a trophic unit, propagating and amplifying the response at the cell surface into the submucosa. In addition to structural damage, the epithelium displays an "activated" phenotype evident by activation of transcription factors such as nuclear factor kappa B (NF kappa B), and expression of mediators which directly or indirectly lead to a chronic cycle of inflammation and injury. A diverse number of innocuous stimuli trigger asthma. It is likely that interactions between genetic and environmental factors converge on common intracellular signalling pathways that regulate epithelial stress and repair. Of particular relevance is the NF kappa B signalling pathway and the mitogen activated protein kinase pathways (MAPKs), of which the mitogen activated extracellular regulated kinases (ERKs), and the stress activated P38 and c-Jun NH2 terminal kinase (JNKs) are best known. This review aims to highlight the importance of these signalling pathways in coordinating the response to diverse stimuli at the surface of the bronchial epithelium which leads to development and maintenance of the asthmatic state. PMID- 11407211 TI - Smoking reduction in smokers compliant to a smoking cessation trial with nicotine patch. AB - The aim of this retrospective analysis was to assess the extent of smoking reduction in smokers who were compliant to a smoking cessation trial with nicotine patch, and failed to completely quit smoking. Out of 297 smokers in total, 237 participants received active treatment (60 received placebo). Eighty treated subjects attended all the scheduled visits and were classified as either abstainers (nonsmokers), regular smokers or occasional smokers. Compared to the remaining 157 participants, these 80 subjects had significantly lower mean baseline daily cigarette consumption (24 versus 30; p < 0.001), expired carbon monoxide levels (25 versus 33 ppm; p < 0.001), plasma nicotine and cotinine levels, and Fagerstrom Tolerance Questionnaire score (5.7 versus 7.0; p < 0.001). All subjects received active treatment for up to 18 weeks (full dose for 12 weeks plus tapering dose for 6 weeks), with follow-up visits scheduled up to 1 yr. A statistically significant reduction in cigarette consumption (versus baseline) was observed among both the occasional (-99%) and regular (-77%) smokers between week 1 and week 52 (p < 0.001). Concomitant smoking and patch use was well tolerated since adverse events were infrequent, mild and transient. Thus, in addition to those subjects who successfully quit smoking, a further group of subjects who attended all the follow-up visits during the smoking cessation trial significantly reduced their mean daily cigarette consumption. PMID- 11407212 TI - Mechanisms and characteristics of airway sensitization to indoor allergens. AB - The increasing prevalence of allergic respiratory diseases, and particularly of bronchial asthma, has been linked to changes induced by human activities in outdoor and indoor environments. People living in industrialized countries spend most of their time indoors: in private homes, offices and means of transport. Indoor environments are not a refuge from outdoor air pollution. Modern systems for energy saving such as insulated windows and doors reduce the indoor natural ventilation and consequently increase the rate of indoor humidity. These conditions may determine an increase in the level of indoor pollutants (tobacco smoke, gases produced by cooling processes etc.) and of allergens derived from mites, domestic animals and cockroaches. Upholstered furniture, wall-to-wall carpets, central heating systems and/or humidifiers may also contribute to the growth of mite populations. The increasing levels of exposure to pollutants and allergens in indoor environments represents a risk factor for the development of airway sensitization, especially if these materials are inhaled early in life. The major cat allergen Fel d 1 is considered an ubiquitous allergen, since it has been found in many indoor environments where a cat has never been kept. The clothing of cat owners seems to help spread Fel d 1 in cat-free environments. Sensitization to cockroach allergens is very common in patients living in urban areas where unhygenic conditions may favour the growth of cockroach populations. Monitoring of the levels of allergens and strategies of allergen and pollutant avoidance in indoor environments are the main ways to reduce the prevalence of respiratory allergies induced by these materials. PMID- 11407213 TI - Excessive daytime sleepiness and the sleep apnoea/hypopnoea syndrome: a major public health problem? AB - There is unequivocal evidence that the sleep apnoea/hypopnoea syndrome (SAHS) causes daytime sleepiness and a consequential increase in road accidents. There is not, however, good agreement about the magnitude of that increase in risk. Estimates for the increased risk range from 1.3 to at least 6 times that of control populations. These figures can be used to provide estimates suggesting that between 1% and 50% of all road accident deaths are due to SAHS, depending on the prevalence of SAHS and the relative mortality of SAHS related road accidents. There is need for further investigation to increase confidence in these speculative figures, but meantime, in comparison to death rates from other conditions, excessive daytime sleepiness in SAHS is not a proven major public health problem. PMID- 11407214 TI - Functional evaluation before and after interventional bronchoscopy in patients with malignant central airway obstruction. AB - Rigid or flexible interventional bronchoscopy is increasingly accepted for treating central tracheal or bronchial obstruction caused by incurable pulmonary malignancy. Immediate and sustained symptomatic relief and improvement of quality of life can be offered to patients suffering from dyspnoea and facing death by suffocation. Selection of patients based on functional aspects is of central importance for success in endoscopic treatment. A thorough workup is conducted with close cooperation between the various specialists involved in thoracic oncology (pulmonologist, oncologist, radio-oncologist, thoracic and upper airway surgeon). Careful history taking, clinical examination, pulmonary function testing (PFT) and radiologic imaging are needed to assess the general condition of the patient and give information of type and stage of the underlying malignancy. In eligible patients, timing and method of the intervention depend on the acuteness of the presentation and the endoscopic assessment of the lesion responsible for the obstruction. Regular postinterventional follow-up should be conducted by the patient's doctor and consist of clinical examination, PFT and chest radiography. Close cooperation with the specialist centre is necessary to refer patients for repeat bronchoscopy and management of suspected recurrences or complications. PMID- 11407215 TI - Comparison of international guidelines on the control and prevention of tuberculosis. AB - Within the last two yrs a number of countries and organisations have published guidelines on the control of tuberculosis. Though all guidelines have the same principals of disease control, namely the identification and treatment of patients with disease, the screening of individuals at high risk and the giving of preventive therapy to selected individuals with latent tuberculosis infection and the protection of the remainder of the population, they differ in the detail of how this may best be achieved. This article attempts to compare the differences and find the way through to the best practice. PMID- 11407216 TI - Towards a tobacco-free world. AB - Many barriers must be tackled for controlling the tobacco epidemic responsible for completely preventable mortality. Tobacco addiction is presently considered as the central element in smoking. The mechanisms of action of drugs of abuse (including nicotine) and the current techniques for their study in animals and in humans are briefly reviewed. Cigarettes are manipulated by the tobacco industry to increase the level of dependence. The transnational tobacco companies use a global tactic to expand their sales, particularly in the third world, so that the mean world cigarette consumption per adult remains unchanged at approximately 1500 per year. Nevertheless, it is hoped that control of the epidemic will be gained in the 21st century, since the awareness of tobacco risks and costs is increasing, an antismoking climate is developing and the tobacco lobby has lost its credibility. On the other hand, improved behavioural and pharmacological methods increase the cessation rate. Health professionals are progressively more involved in the smoking cessation problems, while international control measures are applied by the European Commission. The World Health Organization is preparing a "Framework Convention on Tobacco Control" with binding mechanisms for governments. A larger involvement of the community is needed to promote an international policy based on health values, rather than on trade considerations, and addressing the specific problems of the third world. Nevertheless, since a latency of approximately 30 yrs exists between the increase in tobacco consumption in a population and the rise in tobacco mortality, past trends in consumption will continue to influence health status during the next 30 yrs. Present, urgently needed, radical health measures are expected to have their major impact only after that period. PMID- 11407217 TI - Nitric oxide (NO) synthesis and mechanisms of action. PMID- 11407218 TI - Indirect methods to measure nitric oxide. PMID- 11407219 TI - Direct methods for the measurement of nitric oxide. PMID- 11407220 TI - Electrochemical measurement of nitric oxide. PMID- 11407221 TI - Activity of "NO-donor" or "NO-mediated" drugs. PMID- 11407222 TI - [Allergies: environmental illness no. 1]. PMID- 11407223 TI - [Epidemiology of allergies in Switzerland]. AB - The Swiss Study on Air Pollution and Lung Diseases in Adults (SAPALDIA) was carried out during 1991-1993 in eight Swiss areas with different environmental characteristics. The cross-sectional examination included 9651 adults, aged 18-60 years, who all participated in a detailed interview. In 8357 subjects complete allergy skin and in-vitro tests were available in addition. The prevalence of atopic sensitization (positive skin prick test to any of the tested inhalant allergens and/or a positive Phadiatop as an in-vitro screening test for atopy) was 32.3%, with a higher prevalence in males (35.7%) than in females (28.8%). Skin sensitization was predominantly caused by grass pollen (12.7%), followed by house dust mite (8.9%), silver birch pollen (7.9%) and cat epithelia (3.8%). 11.1% suffered from current hay fever, 6.8% from asthma, 4.5% from atopic asthma. Smokers had statistically significant (p < 0.001) higher mean serum IgE concentrations (geometric mean 39.7 kU/l) than nonsmokers (27.2 kU/l), In Phadiatop positive subjects, the IgE levels were highest, with a mean of 104.3 kU/l (99.0-109.8). The SCARPOL Study (Swiss Study on Childhood Allergy and Respiratory Symptoms with respect to Air Pollution and Climate) ist based on a sample of 4470 children from 10 different areas who completed parenteral questionnaire. 35.7% of the 2879 children who underwent skin prick testing were sensitized to at least one tested aeroallergen, 22.5% to gras pollen, 12.4% to house dust mites, 11.4% to birch pollen and 6.4% to cat epithelia. 17% of the 13- to 15-year-old (8th grade) suffered from hayfever. The prevalence of asthma (ever) for the whole sample was 9%, without differences between the age groups. The lifetime prevalence of atopic dermatitis was 13% and the current prevalence 8%. The risk of eczema was higher in Swiss children than in children of immigrants, in infants with a birthweight below 2500 g, in children with a positive family history of atopic dermatitis, and in children from higher socioeconomic classes. Farm children (n = 133) living in a rural area suffer less frequently from pollinosis (2.4%) and bronchial asthma (1.6%) than children (n = 966) with no direct contact to agriculture, but living in the same area (prevalence of hayfever 18.3%, of asthma 9.1%). This figures are similar to results from former East and Western Germany and from the former USSR and Baltic areas. These large Swiss epidemiologic studies confirmed both, the high prevalence of atopy and atopic diseases, and the health impact of moderate air pollution levels and of factors associated with the 'western lifestyle'. PMID- 11407224 TI - [Allergy and the environment]. AB - The prevalence of allergic diseases has increased in modern Western countries during the last decades. Among the hypotheses for the reasons the idea that environmental pollutants may play a role is intensively discussed. In order to characterise the influence of these pollutants on the development, elicitation and chronification of allergic reactions (allergotoxicology) epidemiological, clinical and experimental data have to be considered. These investigations showed that among the pollutants with an enhancing effect on allergic diseases pollution types with nitrogen oxides (NOx), ozone (O3), tobacco smoke, particulate matter and diesel exhaust particles are of special interest. PMID- 11407225 TI - [The course of allergy: principles for early diagnosis, prevention and early therapy of allergic diseases]. AB - Allergic diseases have a peculiar characteristic: at different ages of the child they present under different forms. The typical time course, where atopic children 'grow out' of one allergic disease (e.g. atopic dermatitis) but then suffer from the next form of allergic disease (e.g. allergic asthma) has been termed 'atopic march', or 'allergic march'. Various information can help to identify early in life those children at risk for engaging in this atopic march. Sensitization to hen's egg and mites during the first year of life, long-lasting sensitization to food allergens during the first two years of age and atopic dermatitis in early infancy have been found to be predictors of allergic sensitizations and/or allergic airway disease later on in childhood. Although today no measures are available to completely prevent any manifestation of atopy before disease onset, some measures can contribute to stop or at least slow down the progression of the allergic march. Breast feeding for a least 4 to 6 months, stepwise introduction of solid food after this age only and avoidance of highly allergenic food during the first year of life can help to achieve this goal, as do measures to reduce indoor allergen exposure (especially house dust mites). For selected patients suffering from atopic dermatitis who are sensitized to grass pollen and/or house dust mites and who have highly motivated parents an early treatment with an antihistamine (cetirizine) can be considered. PMID- 11407226 TI - [Pathophysiological and pharmacotherapeutic aspects of IgE-mediated allergic reactions]. AB - IgE-mediated allergic diseases are associated with a predominant T-helper (Th) 2 activation. Th2 cytokines regulate IgE production as well as production, tissue infiltration and survival of eosinophils. Advances in our understanding of the pathogenesis of allergic diseases have led to new treatment guidelines, especially for bronchial asthma. Current therapies either neutralize a specific mediator (antihistamines, antileukotrienes) or suppress the immune system in an unspecific manner (corticosteroids). New strategies involve both early intervention within the pathogenic mechanism and high specificity by targeting key molecules. It is hoped that it might be possible to develop drugs with higher efficacy and less side effects. There is a particular need for developing drugs for allergic patients who either require high doses of or do not respond to corticosteroids. This review focuses on mechanisms of IgE-mediated allergic diseases and describes where current and new potential drugs interfere within these pathogenic processes. PMID- 11407227 TI - [Principles of specific immunotherapy of IgE-induced allergic reactions]. AB - Allergen-specific immunotherapy (SIT) aims to selectively skew an allergic immune response into a normal immunity. It appeared that the induction of specific anergy in peripheral T cells and reactivation of anergized T cells by microenvironmental cytokines represent two key steps in the mechanism of SIT. In SIT of bee venom allergy the proliferative and cytokine responses were significantly suppressed within seven days, simultaneously with an increase in IL 10 production. IL-10 induces total anergy in T cells by autokrine interaction. In addition, it can counter-regulate IgE and IgG4 synthesis. The addition of blocking anti-IL-10 to stimulated PBMC fully reconstituted the proliferative and cytokine responses in anergized T-cells. Again, particular cytokines are able to reactivate anergic T cells to produce distinct IFN-gamma/IL-2 or IL-4/IL-13 dominated T cell cytokine patterns and direct by this way SIT towards successful or unsuccessful treatment. The suppression of T cells by IL-10 is an active biochemical process, which depends on the interaction of the ligated IL-10 receptor with the CD28 costimulatory signaling pathway in T cells. PMID- 11407228 TI - [Foods as allergy inducers]. AB - Small children are usually sensitized through the gastrointestinal tract to chemically stable food proteins such as milk, eggs, fish and soy. In adults, however, food allergy is mostly mediated by a primary inhalatory sensitization and because of IgE-crossreactivity by a secondary allergy to food allergens. Thus, in adults a completely different pattern of predominant food allergens is encountered. The leading food allergens are fruits and vegetable as a consequence of pollen associated food allergy. This article summarizes clinically relevant information about the most important food allergens. Furthermore the issue of hidden food allergens is stressed. PMID- 11407229 TI - [Pollen as the cause of allergies]. AB - Pollinosis or hay fever is the most common allergic disease in Switzerland. For symptoms during spring pollens of birch and related trees (alder, hazel) and also ash tree are responsible, while hay fever during summer is mainly caused by pollens of grasses, rye and mugwort. These main plant pollen allergens, relevant cross-reactivities with other pollen and food allergens are reviewed in this article. The well-established methods of pollen-counting in Switzerland allow to define the varying amounts of measurable pollen depending on geographic and climatic conditions. Similarly clinical symptoms, the diagnostic work-up of pollen allergies and therapeutic aspects including preventive measures, symptomatic therapy and specific immunotherapy are presented. Finally, occupational and travelling aspects of pollinosis are briefly discussed. PMID- 11407230 TI - [House dust mites and storage compartment mites as allergy inducers]. AB - Perennial respiratory diseases like chronic rhinopathy or bronchial asthma can often be related to an allergy to indoor allergens. Of these, the most relevant allergens are derived from house dust mites. Epidemiologic data show that 8.9% of the general population exhibit a sensitization to house dust mites. Based on a better knowledge of the biology and the living habits of house dust mites effective allergen avoidance measures can be performed which are, however, quite cumbersome and go beyond just encasing mattresses. Of particular relevance is the reduction of the relative humidity below 50%. Relatives of house dust mites are storage mites which are increasingly realized as an important cause of respiratory allergies. The most important is Lepidoglyphus destructor which is dominant in dust probes from Europe and shows crossreactivity with other storage mites. They are found in barns and grain stores and are thus important causes for occupational respiratory diseases in farmers. The existence of storage mites in house dust is connected with damp housing conditions. PMID- 11407231 TI - [Animals and fungi as allergy inducers]. AB - Pets particularly dog and cat are the men's best friend. In the daily practice respiratory allergy to animal proteins are not uncommon and in some areas the frequency is even higher than allergy to house dust mites. In Switzerland nearly half of the households keeps some kind of a domestic animal with cats followed by dogs as the principal pets. Because the exposure to domestic animals is perennial, allergic symptoms such as rhinoconjunctivitis or asthma usually are less acute than due to pollen or an occupational allergen. The best and most effective management of a domestic animal allergy is to avoid having contact with the relevant pet. Because of personal and emotional conflicts other strategies are employed to reduce allergen levels of the pet such as by washing or by restriction of the territory. For many years, fungal spores have been recognized as potential causes of respiratory allergies. Besides the more community recognized microfungi or molds such as Alternaria alternata, Aspergillus fumigatus or Cladosporium herbarum the class of basidiomycetes--physically the largest and morphologically the most complex fungi--are known to produce allergic symptoms. This class consists of about 14,000 species, including mushrooms, bracket fungi, puffballs, toad stools and jelly fungi, as well as the plant pathogenic rusts and smuts. Clinically, symptoms due to fungal allergens are not distinguishable from those due to pollen, however, in recent years asthma attacks particularly in young people have been associated with high spore counts in the air. Upon contact fungal components may cause eczema or trigger inflammatory skin eruptions in a subgroup of patients with atopic eczema. Though food allergies to mushrooms are largely anecdotal, a few well documented cases mainly due to Boletus edulis (king bolete or cepe) have been published recently. Since fungal spores are ubiquitous atmospheric components avoidance measures are often not achievable. Nevertheless, some preventive steps especially at home may help to control growth of molds. Specific immunotherapy may be indicated for both domestic animal and fungal (mold) allergies in certain circumstances. Clinical relevance, however, needs to be demonstrated by an accurate allergological assessment. PMID- 11407232 TI - [Therapy of neurodermatitis (atopic dermatitis)]. AB - Atopic dermatitis is a chronic-relapsing disease that often requires an individualized therapeutic approach. There are various treatment options that address known pathogenetic pathways and, when used in combination, are often very successfull. In view of the fact that a unifying pathogenic concept as well as a therapeutic regimen is not available to this very day, preventative measures that should be introduced in very early childhood. Therefore, one very important goal in treatment is to identify and eliminate triggering factors which include irritants, allergens and cutaneous bacterial, viral or fungial infections. The mainstay of therapy today are external emmollients, which when used appropriately, can reduce the need for systemic treatments. The reconvalescence can be speeded up by auxilliary therapies such as UVA or UVB, psychotherapeutical strategies such as stress reduction as well as so-called climate-("Klima") therapy. PMID- 11407233 TI - [Therapy of allergic rhinitis]. AB - Allergic rhinitis is a common disease with a prevalence of 10-20% in western countries. Allergic rhinitis may be complicated by the possible restriction of quality of life and can lead to sequelae like sinusitis, headache or even allergic asthma. The treatment of allergic rhinitis is mainly based on allergen avoidance, pharmacological treatment and specific immunotherapy. For mild symptoms of seasonal or perennial allergic rhinitis topical or nonsedating second generation oral H1-antihistamines or chromones are advised. If the patient presents symptoms of long duration or nasal obstruction is dominant, intranasal steroids should be used, which have proved to be an effective and safe form of therapy for allergic rhinitis. A combination of oral antihistamines and steroids are possible and recommended if one of these agents alone does not provide sufficient relief. If necessary this regimen is supplemented with topical antihistamines or chromone eyedrops. In cases of severe nasal obstruction, a short course of oral steroids or topical decongestants, which both should not be given longer than ten days, is recommended. Intramuscular corticosteroids should not be given, due to the suppression of adrenal glands. In addition it is important to prevent exposure to the allergen. If the treatment is not effective, further investigations should be done to exclude other nasal diseases (polyposis nasi, anatomical anomalies, chronic sinusitis). This article summarizes the recommended medications with their possible side-effects and their place in therapy management of allergic rhinitis in adults and children. PMID- 11407234 TI - [Therapy of allergic bronchial asthma]. AB - Asthma and atopy are strongly related conditions. The presence of specific IgE to perennial (arthropods, animal dander and other), seasonal (pollens, certain fungal spores) and occupational allergens is associated with the occurrence of asthmatic symptoms. Current therapy is based on combining three principles: Avoidance of trigger factors and of allergen exposure, drug therapy, and specific immunotherapy. Feasability and effectiveness of allergen avoidance (particularly of perennial allergens) have been proven. However, these measures must often be supplemented with drug therapy. Several classes of drugs are nowadays available for treatment of asthmatic symptoms and, most importantly, for control of the bronchial inflammatory process which underlies atopic asthma. Specific immunotherapy is a good treatment option in allergy to pollens, but its use is controversial in allergy to house dust mites and other perennial allergens. Finally, it should be kept in mind that the successful longterm management of patients with atopic asthma depends highly on the compliance of patients. PMID- 11407235 TI - [Therapy of chronic urticaria and recurrent Quincke edema]. AB - A rational and differential therapy of chronic urticaria and relapsing angioedema must be based on a precise classification of the disease. The general management as well as the specific therapy will have to consider the various trigger factors involved, the differences in the response rates to antiallergic drugs and the different prognoses of the urticaria subforms. Therefore this article will first briefly describe the most frequent subforms of urticaria and relapsing angioedema, and then discuss their management and treatment. PMID- 11407236 TI - [Treatment of anaphylactic shock]. AB - Anaphylaxis is a life-threatening allergological emergency: it has to be recognized and treated immediately by the physician. Most often anaphylaxis is caused by drugs, foods and insect stings. Emergency treatment consists in intramuscular application of adrenaline and volume substitution. Only afterwards antihistamines and corticosteroids should be administered. All patients have to be hospitalized immediately and treated on an intensive care unit. After anaphylaxis the following preventive measures are most important: consultation with an allergist, including instruction for allergen avoidance and emergency medications. In anaphylaxis induced by insect stings specific immunotherapy should be started. PMID- 11407237 TI - [Specific and nonspecific (anti-IgE) immunotherapy of allergic diseases]. AB - Specific immunotherapy comprises a special form of allergy treatment which consists of stepwise increasing doses of the allergen, given subcutaneously or orally with the aim to reprogram the specific immunity (from allergy to tolerance). This requires some experience and an exact allergological workup, since the main mistake in the treatment of this specific immunotherapy is the selection of unsuitable patients. The effectivity of specific immunotherapy is well documented for bee and wasp venom allergy, pollinosis and more and more also for asthma bronchiale. A type of unspecific passive immunotherapy is the injection of anti-IgE antibodies. However they block only soluble IgE and are not able to remove mast cell bound IgE. Thus, the allergic reactivity persists. Clinical studies have shown some efficacy in asthma as well as in pollinosis. However, at present it is unclear, whether this drug, which has to be given every two to four weeks in relatively high concentrations, is effective in patients with polysensitization (i.e. patients suffering from mite, fungal and pollen allergy) or is effective in patients which have manifestations of their allergy in different organs. PMID- 11407238 TI - [What is your diagnosis? Basal meningitis and rhombencephalitis. Blood culture positive for Listeria monocytogenes]. PMID- 11407239 TI - [Your ultrasound diagnosis? Acute cholecystitis]. PMID- 11407240 TI - [Rational therapy of Alzheimer dementia based on current clinical studies]. AB - Several preparations are currently available for the treatment of Alzheimer's dementia. To date only some of these drugs could show evidence of efficacy that would justify a recommendation for their therapeutic use. In the present survey, the results of five controlled clinical trials with acetylcholinesterase inhibitors versus Ginkgo-biloba extract have been analysed. The trials conducted with the acetylcholinesterase altogether have a larger patient population, an acceptable significance level of therapeutic effect as well as a lower rate of drop outs. Based on the documentation presented here, only the approved acetylcholinesterase inhibitors are currently to be recommended under strict scientific criteria for use in the treatment of Alzheimer's dementia. PMID- 11407241 TI - [Breast carcinoma: sentinel lymph node--watchman of intact regional lymph nodes]. PMID- 11407242 TI - [St. John's wort: a pharmaceutical with potentially dangerous interactions]. AB - Over-the-counter preparations of St. John's wort are widely used as 'natural' herbal medicine alternative to traditional antidepressants. The antidepressant effect has been shown in numerous placebo controlled studies. The mechanism of action is assumed to be at least in part, similar to conventional antidepressants, due to presynaptic serotonin reuptake inhibition as well as GABA modulation and inhibition of monoaminoxidases. Because of its favorable safety profile compared to conventional antidepressants, the use of St. John's wort preparations has gained high acceptance with doctors and patients. However, any biologically active compound contains a certain risk of untoward effects and/or interactions which often are neither known nor recognised with the use of herbal remedies. Thus, doctors, pharmacists, and patients might feel themselves in false safety. Recently, a variety of case reports of potentially hazardous interactions due to drug combinations with St. John's wort have been published (e.g. cellular rejection of pancreas-, kidney- as well as heart transplants with ciclosporin therapy, rise of INR with oral anticoagulants, bleeding with oral contraceptives, reduction of plasma concentration of digoxin, indinavir, amitriptyline, and theophylline). We report a case of irregular bleeding with oral contraception and discuss these drug interactions and the mechanisms. PMID- 11407243 TI - [Cutaneous listeriosis]. PMID- 11407244 TI - [Thyrotoxic crisis]. PMID- 11407245 TI - [31-year-old patient with right-sided thoracic pain and dyspnea]. PMID- 11407246 TI - [Extracorporal shock wave therapy in veterinary medicine]. AB - This article gives a general overview on a new therapy in equine orthopedics. In human medicine extracorporal shock wave therapy (ESWT) is used for lithotripsy of stones of the urologic system and also in orthopedics. In equine medicine the therapy is almost exclusively applied for the treatment of orthopedic problems. The main indications are proximal suspensory desmitis (PSD), lesions of tendons and ligaments, osteoporotic changes and spavin. In this paper the mode of action of shock waves and the different shock wave devices are discussed. Also, some information about an ongoing clinical trial at the Animal Hospital University of Zurich is given. PMID- 11407247 TI - [Clinical experiences with cytology in the dog]. AB - In a retrospective study (1995) cytological diagnoses of 3987 specimen of different origin in the dog were evaluated. Biopsies were performed at the Department of Clinical Sciences of Companion animals at the University of Utrecht or sent in by external practitioners. Biopsies were obtained from different regions including skin/subcutis, mammary gland, lymph node, external ear canal, kidney, urinary bladder, urethra, prostatic gland, testes, bone, bone marrow, synovia, nose, oral cavity, cerebrospinal fluid, bronchial lavage, lung, spleen, liver, conjunctiva, and retrobulbar region. The majority of biopsies were obtained from skin/subcutis (n = 1815), lymph node (n = 706), and mammary gland (n = 415). A high percentage of non-diagnostic biopsies were observed with biopsies of lung, mammary gland, cerebrospinal fluid, external ear canal, and oral cavity. The frequencies of diagnoses in the different areas will be presented and the diagnostic indications of cytology are discussed. PMID- 11407248 TI - [The use of memoprint in the cat]. AB - In this study, done in the context of normal veterinary practice, the possibilities of using Memoprint have been evaluated. The values of systolic arterial pressure (SAP), diastolic arterial pressure (DAP) and pulse rate (PR) in healthy cats (n = 72) have been determined, as well as the values of SAP, DAP and PR before (n = 26), during (n = 40) and after (n = 16) an anaesthesia for routine surgery. The results show that Memoprint allows a quick, convenient and correct determination of SAP (average: 122.7 mmHg) and DAP (88.5), but an imprecise determination of the PR. The measures are not highly repeatable with conscious cats (SAP: 0.68, DAP: 0.51), unlike that obtained before (0.90, 0.86), during (0.96, 0.90) and after (0.88, 0.58) the anaesthesia, indicating once again the importance of the cats' familiarity and cooperation with the veterinarian. An analysis of the variance of the values of blood pressure shows their direct relationship with age, sex, breed and style of life of the cats and an increase in pressure in those over ten years of age. PMID- 11407249 TI - [Three cases of postweaning multisystemic wasting syndrome (PMWS) due to porcine circovirus type 2 (PCV 2) in Switzerland]. AB - Porcine circovirus type 2 (PCV 2) was found in three five- and nine-week-old pigs from two feeder pig producer farms. Clinical signs were persistent diarrhea and wasting. The animals showed histomorphologic changes characteristic for "Postweaning Multisystemic Syndrome" (PMWS). One animal had the typical basophilic intracytoplasmatic circoviral inclusion bodies in the Peyer's patches of the ileum. Circoviral DNA was detected in the lymphatic organs as well as in the liver. This case report is the first description of PCV 2 in Switzerland. PMID- 11407250 TI - [Life threatening metabolic disorders after application of a sodium phosphate containing enema in the dog and cat]. AB - Application of a sodium phosphate containing enema caused life-threatening metabolic disorders in a Dachshund and two cats. Clinical signs were characterised by dehydration and various neurological deficits including seizures. Most striking laboratory abnormalities were hypernatremia, hypocalcemia, hyperphosphatemia, and polycythemia. Despite intensive treatment the dog died, whereas the cats recovered completely. In face of possible severe and potentially fatal metabolic abnormalities sodium phosphate enemas, such as Practo-Clyss, should be used with caution or not at all in cats and small dogs. PMID- 11407251 TI - [Unenlightened psychiatrists, psychologists, caregivers, social workers, police and politicians]. PMID- 11407252 TI - [Stress of family caregivers of psychiatric patients. Developmental trends, concepts and results of research]. AB - OBJECTIVE: A review of the findings in the research literature on the effect of mental illness on other members of the patient's family is provided. This article critically examines the research to date with special reference to the conceptual and operational definitions of burden and methodological issues. RESULTS: The burdens of caring for a patient at home are considerable throughout different patient groups. Relatives' experience of burden is influenced by their cognitive appraisal of the situation and their psychosocial resources of coping with it. Research findings suggest that information and support provided by clinicians can alleviate caregiver burden, although it has been used too rarely in clinical routine. CONCLUSIONS: Finally, the review identifies advances made in this line of research in recent years and highlights areas that need further attention in future research work. Systematic research of families' responses to and management of mental illness should move beyond the more global concept of family and address the great diversity among families with mentally ill relatives. Furthermore, given the lack of consensus regarding the theoretical conceptualizing of the burden concept a new framework with reference to stressrelated research is suggested. PMID- 11407253 TI - [Family consultation as a part of routine management in a psychiatric and psychotherapy department of the general hospital]. AB - BACKGROUND: Covers two years experience of family visits (FV) as a routine fixture at a psychiatric ward in a General Hospital. PATIENTS AND METHODS: In the course of one year 98 visits took place involving a total of 481 contacts between therapists, patients, and relatives. 269 patients participated up to 10 times in FV. A total of 666 family members participated, the bulk of them partners, mothers and children. Each FV contact is restricted to 20 minutes, but can be repeated as often as desired. An ever recurrent theme is the question family members ask as to the causes of the patients condition and how they can expedite the healing process and ensure compliance. The 269 FV patients do not differ markedly from the other patients concerning age, sex, diagnosis and treatment time. CONCLUSION: FV are experienced by patients and relatives as positiv and enriching. For psychiatric workers an ideal way to learn the psychotherapeutic rudiments. PMID- 11407254 TI - [Family information days. Attitudes and needs of family of schizophrenic patients]. AB - OBJECTIVE: This study aimed to examine attitudes, knowledge about disease and needs of relatives of schizophrenic patients. METHOD: Questions concerning needs and wishes addressed to professionals were added to the "Questionnaire of attitudes concerning disease for relatives". The questionnaire was distributed to relatives participating in the workshop at Bezirksklinikum Regensburg and sent by post to non-participating relatives. RESULTS: The relatives of both groups were predominantly comparable regarding attitudes, knowledge and needs. Mainly they wished more dialogue with therapists. Information about schizophrenia was assessed as very important. Relatives participating in the workshop reported significantly more frequently that the atmosphere in the family is strained and that they understand the nature of the illness less than non-participating relatives. They reproached themselves more frequently, but felt less depressive than non-participating relatives. 95% of the participating relatives were satisfied or very satisfied with the workshop. They assessed self-help groups for relatives of schizophrenics as more important than non-participating relatives. CONCLUSION: Workshops for relatives of schizophrenic patients have proved to be economic and well accepted to contact many relatives and to give attention to their problems. PMID- 11407255 TI - [Mother-child treatments within the scope of public community psychiatric care. Clinical treatment concept and experiences over 5 years]. AB - BACKGROUND: Described are mother-child treatments performed within the framework of regional maximum care at general psychiatric units not having such a treatments focus. PATIENTS AND METHODS: The new clinical concept with its social and psychodynamic aspects is described. It provides realistic perspectives for all involved. Last but not least it puts the child in a position as a helpful companion for the mother. Over a space of 5 years, 20 mother-child treatments were performed at four different wards. 12 mothers suffered from schizophrenia (F20), 7 had depressive complaints (F32, F43), and one patient had a borderline personality disorder (F60). Treatments lasted an average of 10 weeks, all mothers retained their children. 16 patients were sent home; 2 were referred to a mother child facility; 1 patient was placed in a half-way home and another in a house for women, in both cases with offspring. CONCLUSION: Mother-child treatment under specific spatial conditions is not only possible, but is an enrichment for all at general psychiatric units attending heterogeneous patients. The main requirement is a certain level of treatment care, such as an established primary care track, a clinic team that is flexible and open to novel approaches and, as a result, willing to accept a certain level of stress as normal. PMID- 11407256 TI - [Ego identity and self-concept problems in children of psychotic parents. Results of a qualitative cross-sectional study]. AB - OBJECTIVE: In contrast to previous high-risk studies of children of parents suffering from psychosis a new approach to the difficulties of affected children will be tested, using the linguistic and narrative tools of biography studies. METHODS: The transcribed texts of biographical interviews of affected children will be analysed linguistically and hermeneutically through "close reading". RESULTS: The linguistic and interpretative study of these texts will demonstrate the children's specific problem of identity-formation and their construction of reality and biography through the order of language (as shown by their uses of metonymy and metaphor). CONCLUSIONS: By using the method of biographical research a) the creative potential of patients and their children is exposed, b) a qualified view of mental illnesses can be reached in the field of symbolic practice. PMID- 11407257 TI - ["The illness has totally changed our lives"--stress in partners of schizophrenic patients at the onset of illness]. AB - OBJECTIVE AND METHOD: The aim of this study was to investigate how spouses of schizophrenic patients experience the onset of the disease. Based on the narrative analysis of 23 in-depth interviews, the article highlights different aspects of this stressful life event. RESULTS: The first episode of a schizophrenic disorder leads to severe distress in the spouse of the patient. Emotions of fear, despair and loss of control are especially pronounced if spouses feel that neither their information about the disease nor their supportive resources are sufficient. The illness is often viewed as an existential threat to marriage and family life. DISCUSSION: Supportive services for spouses of schizophrenic patients should be offered very closely to the onset of schizophrenia, because the situation is especially burdensome in this period. Interventions should meet the particular needs of spouses. In addition to information about the illness and coping strategies intervention programs should consider issues which are relevant for partnership and parenting roles. PMID- 11407258 TI - [Etiology of schizophrenia from the viewpoint of family]. AB - OBJECTIVE: In a former quantitative study on relatives' beliefs about the causes of functional psychoses a preference for psychosocial explanations was found. There were also indications of an inclination towards a multicausal concept. The aim of this study is to try to replicate these findings using semi-qualitative methods. In addition, we are interested in knowing to what extent the causal beliefs of relatives are similar to those of the patients. METHOD: Problem centered interviews were conducted with 31 relatives of schizophrenic patients. A computer-assisted qualitative content analysis was carried out with the transcripts of the interviews. RESULTS: As in the previous study, relatives showed a strong propendency to endorse psychosocial explanations. However, among only one third a multicausal concept could be observed. The causal beliefs of relatives and patients were quite similar. DISCUSSION: The findings are discussed with reference to the concept of subjective illness theory and the concept of social representations. PMID- 11407259 TI - [Attitude of family of psychiatric patients to psychiatric research, especially to early detection of schizophrenic psychoses]. AB - OBJECTIVE: To assess attitudes of relatives of mentally ill patients to psychiatric research, especially to early detection of schizophrenic psychosis, and to evaluate the potential stigmatization effect of an early diagnosis of schizophrenic psychosis. METHODS: A questionnaire was sent to relatives of mentally ill patients, all members of a self-help group in Switzerland. They were asked to answer structured questions about their personal attitude to psychiatric research in general, their willingness to support an affected person to participate in a research project, their attitude to early detection of schizophrenia and to the potential stigmatization of an early diagnosis. RESULTS: 88% of the relatives (n = 200) have a positive attitude to research in psychiatry. 47% would support their affected family member to participate in a research project whereas 39% would not. 52.5% fully share the expectation of psychiatric research to attain a better course by earlier detection of psychosis, 24.5% tend to agree. 20.5% of the respondents indicate a fear of stigmatization for the affected by earlier diagnosis, while 12% fear stigmatization for themselves. CONCLUSIONS: A surprising fact is the positive attitude of relatives to psychiatric research in general, but also to early detection of schizophrenic psychosis. But this cannot lead to the conclusion that relatives are supporting the participation of an affected family member in research. The small number of relatives that fears a stigmatization by earlier diagnosis of schizophrenic psychosis shows that stigmatization depends on other factors than the diagnosis, i.e. psychiatric hospitalization or medication side effects. These findings encourage psychiatric research to increase knowledge of the early phase of schizophrenia in favor of patients and their relatives. PMID- 11407260 TI - [Clinic admission?--but not without my child!]. AB - Since the end of the 1990s the Westfalische Klinik Lengerich has offered an inpatient psychiatric treatment of an psychiatric ill parent--combined with a pedagogic attendance of the child taken along. The following article describes our experience in that matter, from a ward concentrated on psychotherapy. In selected examples supposed patterns of interaction--following the specific disease--are represented from both the psychotherapeutic und the pedagogic point of view. In practice our plan of treatment and attendance has proved successful for the wellbeing of the patients and their children. Remains to be hoped that- despite difficulties in organisation which will have to be overcome--similar offers can be made in regional psychiatric attendance. PMID- 11407261 TI - [Viewpoint of family: the black hole in the social psychiatric system]. AB - The following review concludes the authors experience of a 6 year consultation with the parents of a 28 year old daughter with a schizophrenic psychosis. The self-sacrificing experiences of the parents are reported as well as the experiences with a well developed system of psychiatric care with several professionals, whose work remains without success because of insufficient coordination. The author asks for a conference of all involved persons, to nominate a responsible therapeutic key-worker to coordinate the different psychiatric care in the future. PMID- 11407262 TI - [Characteristics of families of women with restricting anorexia nervosa compared with families of normal probands]. AB - This study compares families of women with restricting anorexia nervosa with families of women without any psychiatric history on the following variables: parents' eating problems (Eating Attitudes Test: EAT-26) and psychopathology (Symptom Checklist-90-R: SCL-90-R); family members' perceptions of the parent daughter relationship (Parental Bonding Instrument: PBI) and the family's overall functioning (Self-Report Family Inventory: SFI). Twenty-eight young women suffering from restricting anorexia nervosa, 27 non-clinical women and their respective parents, completed these questionnaires and also participated in a semi-structured individual interview. Our results demonstrated that parents in the two groups had similar eating attitudes and behaviors. However, parents of women with anorexia nervosa reported more problems with alcohol consumption than non-clinical parents. There were no differences between the two groups on the measure of family functioning (SFI), all scores in the families of anorectics being within normal limits. All the daughters, however, reported less family cohesion and more family conflict than did their parents. On the PBI, all the daughters reported receiving more care from their mothers than their fathers; the anorectic women reported experiencing more maternal control, intrusiveness and overprotection. The relationship between each family member's perception of family functioning and the parent-daughter relationship was examined. Anorectic daughters associate the emotional health of their family with the care received from both parents, whereas control women associate this variable with their father's attitudes and behaviors. However, fathers from both groups do not consider their own care or protection as having contributed to the health of the family. PMID- 11407263 TI - [Symptoms of DSM IV borderline personality disorder in a nonclinical population of adolescents: study of a series of 35 patients]. AB - 1,363 high school students were solicited to complete a personality disorder questionnaire and were encouraged to continue in the study by signing up for interviews with Master's level psychology students. 107 students (7.8%, 34 males, 73 females, mean age = 16.7 +/- 1.8) manifested themselves for the interview and were assessed by using structured diagnostic interviews for borderline personality disorder and major depressive disorder (DIB-R, Revised Diagnostic Interview for Borderlines; MINI, Mini International Neuropsychiatric Interview). The interviews were audiotaped. Interrater reliability was determined by independent ratings of 12 borderline subjects and 12 non-borderline subjects (kappa: 0.795). The distribution of the 107 subjects based on the number of DSM IV borderline personality disorder criteria indicated a gradual dispersion suggesting a continuum from normality to borderline personality disorder: 8% of the subjects met none of the criteria; 16% met one criterion; 17% met two; 12.5%, three; 13.7%, four; 8.4%, five; 5.6%, six; 9.3%, seven; 4.6%, eight; 4.6%, nine. Thirty-five of these 107 subjects (32.7%, 6 males, 29 females, mean age = 16.7 +/ 1.7) received a diagnosis of borderline personality disorder according to DSM IV criteria. The most frequent symptoms were paranoid ideation or dissociative symptoms (97.1%), affective instability (88.6%), inappropriate, intense anger (85.6%), suicidal gestures or automutilation (82.9%), followed by frantic efforts to avoid abandonment (77%), impulsivity (65.7%), unstable and intense relationships (62.9%), identity disturbance (60%), and emptiness (57.1%). The comparison between borderline and non-borderline subjects showed that all borderline personality disorder criteria discriminated significantly between the two groups. The high incidence of paranoid ideation (97.1%) and dissociative experiences (65.7%) in the borderline group suggests the pertinence of criterion 9 in the diagnosis of borderline personality disorder in adolescents. Two criteria of schizotypal personality disorder were also frequent in this group: 68.6% of the borderline group reported odd beliefs or magical thinking, in particular beliefs in clairvoyance or telepathy and 88.6% reported unusual perceptual experiences, in particular sensing the presence of a force or person and bodily illusions. Moreover, 31.4% of the borderline group reported transient "quasi" psychotic experiences, mainly "quasi" visual hallucinations. Auditory hallucinations or delusional ideas were not observed. This symptomatology suggests a "quasi" psychotic dimension of adolescent borderline personality disorder. Affective instability was the next most frequent symptom which was usually marked by a cyclothymic appearance. Comorbidity with major depressive disorder was high: 85.7% of the borderline subjects had a concurrent diagnosis of major depression versus 45.8% of the non-borderline subjects. Thus, major depression is more frequent than most of the borderline personality disorder criteria, with the exception of the already noted paranoid ideation and affective instability. Hypomanic symptoms were frequent in the borderline group (65.7%) as well as in the non-borderline group (38.8%). This symptomatology suggests that adolescent borderline personality disorder is linked to an attenuated bipolar spectrum characterised by major depressive episodes and soft signs of bipolarity. However, hypomanic symptoms, which were quite frequent in non-borderline subjects, might also be due to a mechanism of defence, i.e. the denial of depression. Comorbidity with anxiety disorders appeared also to be high: anxiety symptoms were found in 91.4% of the borderline subjects who reported symptoms of generalised anxiety disorder, panic disorder, and somatoform disorders. The overall clinical appearance of these borderline adolescents not referred for treatment seemed to be quite similar to that of borderline adolescents in clinical samples. This study shows that adolescent borderline personality disorder in non-clinical population is a serious disorder characterised by the importance of mental suffering and behavioural disturbances the disorganising power of which may fix the developmental process in a pathological pathway. Adolescent borderline personality disorder appears in this study to be strongly associated with major depressive disorder and at-risk behaviours linked to impulsivity, affective instability, and suicidal ideation. However, this study found an absence of precise cut-off between borderline and non-borderline subjects. Two factors might have contributed to the appearance of a continuum. First, some degree of impulsivity and instability in affectivity, self-images and interpersonal relationships is part of normal adolescence. (ABSTRACT TRUNCATED) PMID- 11407264 TI - [Current status of electroconvulsive therapy in adult psychiatric care in France]. AB - A survey into the practice of electroconvulsive therapy between November 1996 and November 1997 in all the French Psychiatric Public Hospital Services is set out here. Of the 815 questionnaires sent by mail, the authors mention that 48% replied. Among the 391 State Hospital Services which responded, 51% declared having practiced ECT during that period of time. A detailed analysis is supplied regarding the apparatus type, maintainance ECT, anaesthesic used, electrode positioning, medical indication and side effects. The results are presented, then discussed one by one before being compared with those of a similar study made ten years ago. We can already point out to a significant decline in ECT from the 64% who affirmed having practiced it in 1986 to the 51% the authors found in their survey. This undeniable fact has however to be counterweighed against the marked increase in maintainance ECT (40% against 11% with p < 0.001) and the qualitative improvement of the practical application of that therapy if one has to judge from the modernisation of the apparatus as well as the move towards better security conditions. A tentative evaluation of the impact of the guidelines described in the French Government November 1996 circular is also made. Last but not least, the proper equipment for practising ECT seems to be sadly lacking in many of the Psychiatric Wards. This brings further to the fore the debate, so common in France, regarding the equal distribution of the availability of all medical care to each and every citizen wherever he may be living. PMID- 11407265 TI - [Depression, anti-thyroid antibodies and Hashimoto encephalopathy]. AB - Psychiatric manifestations are infrequent or rarely described in Hashimoto's encephalopathy. It usually begins like a subacute diffuse encephalopathy with confusion, tremor and other neurologic symptoms. A relapsing course is characteristic. Neither biologic nor clinical symptoms are specific but high antithyroid antibodies levels are characteristic. The diagnosis can be seriously delayed by the fact that the different symptoms implicate approaches by psychiatrists, neurologists or endocrinologists. There are two clinical types. The one presented here evaluates progressively to dementia with psychotic episodes, confusion and seizures. An early steroid treatment makes the symptoms regress without aftereffects. We have analysed the clinical and biological findings of a woman who has been admitted to different neurologic and psychiatric departments before her diagnosis was made. First clinical presentation and evolution were that of a depression. Each time the antidepressive treatment was stopped, depression relapsed in spite of an appropriate steroid treatment. Literature shows that a close link exists between depression and antithyroid antibodies whatever thyroid status. This link does still exist after adjustment of psycho-social determinants of depression. The decrease of those antibodies only reflects the decrease of inflammation. So, for the psychiatrist it is important to diagnose Hashimoto's encephalopathy without delay, especially when psychiatric manifestations are in the foreground. Furthermore, a psychiatric report should systematically be added to the clinical and biological findings in order to make a better approach of the existing links between depression and other manifestations of the disease. PMID- 11407266 TI - [French validation of a Minor Morphologic Anomalies Scale in schizophrenic patients and their parents]. AB - Minor Physical Anomalies represent valuable indices of disturbance in early neurodevelopment. They are frequently observed in individuals with various brain disorders, including mental retardation, autism, epilepsy, hyperactivity, foetal alcohol syndrome and schizophrenia. The high prevalence of Minor Physical Anomalies in schizophrenia provides considerable support for a neurodevelopmental model in this disorder. However, studies in large sample using standardised scale are lacking. Such studies are needed in order to confirm their actual frequency and study the clinical correlates or morphological anomalies. OBJECTIVE: The aim of this study was to revise and validate a French version of a scale designed for the evaluation of Minor Physical Anomalies in adult psychiatric patients and notably in patients with schizophrenia. METHODOLOGY: The scale was revised from the Waldrop scale. The choice of items was done on the basis of frequency, reliability in the adult, reliability of rating. Some new items, related to know syndroms with comportmental symptoms were added. Both raters had previously had a short initiation to the rating of the scale. Interrater reliability between two examiners, blind with regards to the diagnosis was evaluated. RESULTS: The interrater reliability was good, with an intraclass correlation coefficient at 0.97. Patients had significantly more minor physical anomalies than comparison subjects, and also more Minor Physical Anomalies than their parents. Fathers and mothers of these schizophrenic patients had significantly more Minor Physical Anomalies than normal comparison subjects. CONCLUSION: Although the evaluation of physical anomalies relies on subjective appreciation of normal vs abnormal, the revised version of minor physical anomalies scale (French version) was found to be a reliable tool, provided that a short initiation to the rating is performed. The scale differentiated schizophrenic patients from their parents, and the latter from the normal controls. A lot of questions remains unanswered concerning the neurodevelopmental hypothesis of schizophrenia. This scale appeared as a useful complementary tool to help in the determination of the developmental phenotypic status of the patients enrolled in pathophysiological studies aiming the identification of developmental factors in schizophrenia. PMID- 11407267 TI - [Frequency and clinical aspects of bipolar II disorder in a French multicenter study: EPIDEP]. AB - This paper presents the definite data from a French multi-center study (EPIDEP). The aim of EPIDEP was to show the feasibility of validating the spectrum of soft bipolar disorders by practicing clinicians. In this report we focus on data concerning the frequency of BP-II disorder and the key characteristics of BP-II by systematic comparison versus Unipolar depression. EPIDEP involved training 48 french psychiatrists in 15 sites; it is based on a common protocol following the DSM IV criteria (Semi-Structured Interview for Hypomania and Major Depression), and Akiskal (Soft Bipolarity), as well as criteria modified from the work of Angst (Hypomania Checklist), the Ahearn-Carroll Bipolarity Scale, HAM-D and Rosenthal Atypical Depression Scale; Semi-Structured Interview for Affective Temperaments (based on Akiskal-Mallya), self-rated Cyclothymia Scale (Akiskal). Comorbidity and family history (Research Diagnostic Criteria) were also obtained; EPIDEP was globally scheduled in two phases: Phase 1 devoted to recruiting major depressives, and phase 2 involved in more sophisticated assessment of soft bipolarity and administrating related measures. Results are presented on the total of 537 patients included at "visit 1" and 493 assessed for soft bipolarity at "visit 2". The BP-II global rate which was 21.7% at initial evaluation, nearly doubled (39.8%) by systematic evaluation of hypomania. Intergroup comparison versus unipolar depressives showed the following key characteristics of BP-II disorder: 1) distinct clinical presentation at index depressive episode despite uniformity in global intensity of depression (overrepresentation in BP-II of "suicidal thoughts", "guilt feelings", "depersonalisation-derealisation", "hypersomnia" "and weight gain"; and of "psychic anxiety" and "initial insomnia" in UP); 2) different course of illness with younger age of onset of first depression, higher rate of suicidal attempts, recurrency and hospitalisations; 3) more difficulties for recognition of the correct diagnosis; 4) more complex temperamental dysregulations (mixture of cyclothymic, hyperthymic and irritable traits which are highly represented in BP-II group); 5) higher rate in family history of mental disorders, especially bipolar disorders. Finally, EPIDEP data confirmed the diagnostic reliability of self-rating of hypomania and cyclothymia. With a systematic search of hypomania, almost 40% of major depressive episodes seen in psychiatric settings were classified as BP-II, of which only half were recognized by the clinicians at study inclusion. The BP-II validity as a distinct disorder from Unipolars was confirmed. Moreover, EPIDEP emphasized the reliability of self-rating in assessing soft-bipolarity (hypomania and cyclothymia). In total, EPIDEP data indicated that recognition of BP-II is feasible in diverse practice settings and proposed for clinicians some adapted clinical tools for assessing soft bipolarity. PMID- 11407268 TI - [Post-traumatic stress, post-traumatic depression and major depressive episode: literature]. AB - Although they are likely to add their effects, physical and psychic traumata (or traumas) can provoke in different ways the appearance of depressive symptoms sometimes common. Post-traumatic depression, reactional depression, major depressive disorder and post-traumatic stress disorder represent different clinical and nosographic disorders in despite of their occasionally common symptomatic core. Historically, it is interesting to note during the XXth century the true semantic change of the terms of trauma from the somatic field to the psychic sphere. Physical traumatism is often represented by a material shock for the subject and by its organic consequences. It is defined as an event that leaves its mark which itself inflicts and handicaps the vital trajectory of the subject. It primarily comprises brain and rachis injuries, whose evolution is frequently characterized by the occurrence/appearance of a depressive disorder, whose genesis rests on psychological but also neurobiologic and physical arguments. Thus major depressive disorders are often present in the course of various physical traumatisms mainly related to nervous system. In accordance with several studies, the prevalence of major depressive disorders ranges from 25% to 50%. These mood disorders occur in the year which follows the accidental event. Their average time of revelation is estimated at four months and their average duration lies between three and six months. Lastly, although these depressive illnesses present clinical symptoms comparable with those observed in other contexts, some nuances can be raised. Nonetheless, they confine sometimes with true clinical forms depending on the intensity, the form, the circumstances or the consequences of the trauma. Psychic traumatism doesn't have the same profile and rests for much dedicated with the reexperiencing. Thus for some authors, depression illness represents a disorder that occurs after a traumatic event whereas others see a differential diagnosis which exludes or which represents a comorbidity with post-traumatic stress disorder. The review of the literature allows us to emphasize the complexity of the links as well as the clinical and epidemiologic differences between stress disorder and major depressive disorder. From the clinical point of view, the major features of PTSD are articulated around a triad of symptoms. They include the reexperiencing symptoms of the traumatic event such as intrusive memories and recurrent nightmares, the protective reactions such as avoidance of the stimuli associated with the trauma and emotional numbing, and the arousal symptoms such as the startled response and hypervigilance. The complexity of this syndrom is due to the frequent combination of these symptoms with other nonspecific ones. As far as the mood is concerned (the mood symptoms are concerned), the regrouping of some of these symptoms allows the clinician to sometimes releave a depressive symptomatology without being able to assess the DSM diagnosis of major depressive disorder. Epidemiologic studies dealing with the risk of installation of a PTSD after a traumatic event reveal differences in the prevalence depending on the nature of the traumatic events: ranging from 1% in general population to 80% following some situations of extreme and durable psychic suffering. Between both poles, one finds a prevalence ranging between 20 and 50% following other events such as serious accidents, natural disasters or criminal assaults. The clinical features of depressive episodes comorbid or associated with PTSD have some characteristics making it possible to individualize various clinical forms as a function of traumatic event type: asthenic, characterial or with somatic symptoms. According to the majority of authors, the co-occurrence of post-traumatic stress disorder and major depressive disorder is high although differential diagnosis is sometimes difficult. However, conceptual differences remain and two conceptions are distinguished. For some authors, like Bleich and Shalev, there would not be true chronological evolution from PTSD to MDD. Moreover the presence of symptoms considered as pertaining to the mood register within the criteria of PTSD would be clearly predictive of the occurrence and the severity of the diagnosis but not of the chronicity. For others, there would be a continuity between post-traumatic stress disorder and major depressive disorder. It is the case in many studies of veterans but also for civilian traumatic events. It is also the case for the American national study of comorbidity in which Kessler concludes that for 78% of the subjects who present a comorbidity PTSD/MDD (comorbidity raised for 48% of the 5,877 subjects included), the mood disorder is secondary to PTSD. (ABSTRACT TRUNCATED) PMID- 11407269 TI - [Efficacy of clozapine in the course of Huntington chorea: apropos of a clinical case]. AB - Previous case reports suggest the potential benefits of use of clozapine on psychotic symptoms in Huntington's chorea. Clozapine was administered to a 70 years old female patient admitted in the inpatient unit of the Department of Psychiatry for paranoid delusions, hallucinations, comorbid with Huntington's chorea. Clozapine was administered with increasing regimen ranging from 25 to 200 mg/day after all other medications were discontinued. No other drugs were associated. Clinical evaluation was carried out at days 0, 28, 56, 112, 280 using the PANSS and Psychotic Depression Rating Scale. The clinical improvement was associated with plasma levels of clozapine which have been recognized as in therapeutic ranges by previous studies. Clozapine proved efficient on both positive and negative symptoms as assessed by a decrease of the PANSS ratings ( 44%) between day 0 and day 56, (-61%) between day 0 and day 280, and by a decrease of Psychotic Depression ratings (-43%) between day 0 and day 56, (-60%) between day 0 and day 280. It was also effective on choreiform movements. The clinical improvement was associated with plasma levels of clozapine which have been recognized as in therapeutic ranges by previous studies. A lower daily dose than those classically used in schizophrenia was sufficient to obtain efficient treatment. PMID- 11407270 TI - [Vulnerability to schizophrenia: neuropsychological performance and schizotypal personality traits]. AB - Although some neuropsychological deficits and a high rate of schizotypal personality disorders have been found in the first-degree relatives of patients with schizophrenia, few studies have looked for a link between those two types of potential marker of vulnerability to this disease. The aims of this study were: 1) to confirm some executive/attentional deficits in a group of first-degree relatives including not only siblings but also parents; 2) to evaluate the schizotypal traits using the French version of 4 self-reporting scales proposed by Chapman and his colleagues; 3) to look for a dependence or independence between the neuropsychological performance and the scores on the scales of schizotypy. Twenty four patients with schizophrenia, 48 of their first-degree relatives and 31 controls were included in the study. Both attentional tests (a Digit Symbol Substitution Test and a Degraded Stimulus-Continuous Performance Test) confirmed a worse performance in the patient and in the first-degree relative groups than in the control group. On the opposite side, the executive performance assessed by the Wisconsin Sorting Card Test, was poorer in the patient group only. Scores of the first-degree relative group on the social anhedonia, physical anhedonia and perceptual aberrations scales were at an intermediate level between those of the patient and control groups; moreover, only scores on the social anhedonia scale tended to be significantly higher in the first-degree relative group than in the control group. Among the first-degree relative group, the only significant correlation found was between the number of perseverative errors on the WCST and the scores on the physical anhedonia scale. PMID- 11407271 TI - [Is cocoa a psychotropic drug? Psychopathologic study of a population of subjects self-identified as chocolate addicts]. AB - The aim of this work was to search for eating disorders, DSM III-R Axis I mental disorders, personality disorders, and addictive behavior, in self-labeled "chocolate addicts". Subjects were recruited through advertisements placed in a university and a hospital. Fifteen subjects were included, 3 men and 12 women aged between 18 and 49. Most of them were not overweight, although 7 thought they had a weight problem. They consumed an average of 50 g per day of pure cacao and, for 13 subjects, this consumption was lasting since childhood or adolescence. The psychological effects of chocolate, as indicated by the subjects, consisted in feelings of increased energy or increased concentration ability, and in an anxiolytic effect during stress. Seven subjects described minor withdrawal symptoms. None of the subjects reached the thresholds for eating disorders on the EAT and BULIT scales. The structured interview (MINI) identified an important ratio of subjects with a history of major depressive episode (13/15), and one woman was currently experiencing a major depressive episode. Four people suffered, or had suffered from anxiety disorders. Although only one subject satisfied all criteria for a personality disorder on the DIP-Q, seven displayed some pathological personality features. The self-labeled "chocoholics" do not seem to suffer from eating disorders, but may represent a population of psychologically vulnerable and depression--or anxiety--prone people. They seem to use chocolate as a light psychotropic drug able to relieve some of their distress. The amount of cacao consumed, although very chronically, remains moderate, and they rarely display other addictive behaviors. PMID- 11407272 TI - [Ultra-rapid detoxification of opiate dependent patients: review of the literature, critiques and proposition for an experimental protocol]. AB - Ultra-rapid opioid detoxification (UROD) is an increasingly popular technique for detoxifying patients addicted to opiates. This technique aims at reducing not only the duration but also the intensity of withdrawal by using general anesthesia coupled with a naloxone or naltrexone medication. In this paper the authors attempt to review the history of UROD and the logic of its procedure and results whilst also demonstrating its advantages and limits. METHOD: The MEDLINE database was searched from 1966 to 2000 using the terms "ultra-rapid opioid detoxification, rapid opioid detoxification under anesthesia, naloxone, naltrexone, opioid-related disorders". Additional data sources included bibliographies in textbooks on substance abuse. RESULTS: Nine studies identified in our search were analysed. The technique is based on a three-phases procedure. It consists of a medical and psychiatric selection of patients addicted to opiates, followed by the detoxification itself and finally a medical and psychosocial follow-up. A brief presentation is made of the theoretical aspects based on the use of a specific opioid receptor antagonist (naloxone and naltrexone). Only inpatients were included in the studies. The detoxification and anesthesia protocols varied. In every study all the subjects were completely detoxified. Only three studies included a control group and two used a randomized design. Three studies reported a follow-up beyond 30 days. DISCUSSION: Although this technique constitutes a safe and effective solution for opiate addicted patients, there are criticisms to be made. The absence of an animal model prior to the study of a human model, the lack of comparison with other procedures, the limitation of available literature, the cost and the risks of this technique and the lack of long-term treatment outcomes obtained from rigorous clinical trials, all call for further assessments. A more rigorous protocol based on the main areas of criticism is proposed (presentation of the inclusion and exclusion criterias, description of the three preliminary interviews, presentation of the UROD technique itself and finally a detailled nine month follow-up). CONCLUSION: Ultra-rapid opiate detoxification represents a potentially safe and effective treatment for opiate addicted patients but more rigorous research methods are needed to render this procedure entirely valid. PMID- 11407273 TI - [Psychiatric manifestations of a new variant of Creutzfeldt-Jakob disease. Apropos of a case]. AB - The new variant of Creutzfeldt-Jakob disease (nvCJD) was first described in the UK in 1996 (16). The nvCJD differs from sporadic, genetic and iatrogenic CJD. Creutzfeldt-Jakob disease is closely associated with an abnormal isoform PrPSc of a cell-surface glycoprotein, prion protein (14). Molecular analysis suggests that nvCJD is caused by the same prion strain as bovine spongiform encephalopathy (BSE) (4, 10). To the end of September 2000, there have been 82 cases of nvCJD in the UK. We report the second French case of nvCJD to our knowledge (5, 13). CASE REPORT: This 36 year old woman was referred by a local general practitioner with a 6 month history of psychiatric symptoms of major depressive disorder. According to her family, the patient had suffered from personality change for several months before the onset of depression including apathy, emotional lability, infantile affect. There was no history of health problems. As she was admitted to the psychiatric department of our hospital in Paris suburbs, she presented a major depressive disorder. There were no specific psychiatric features allowing distinction from common depressive disorders, except a marked emotional lability. The patient's condition progressed rapidly within the following days. She presented memory impairment and disorientation. Drug treatments, clomipramine (125 mg/day) and venlafaxine (200 mg/day), were used with no benefit. She presented subsequently transient delusions and auditory hallucinations, fleeting for some hours. The predominant delusional themes were somatic type and pregnancy. The delusions were concomitant with delusions of the onset of cognitive impairment. The patient tested negative for the P 14.3.3 protein in the CSF. Computed tomography scan of the brain did not show any relevant abnormality. The electroencephalogram showed non specific slow wave activity. The neurological symptoms developed 7 months after the onset of depressive symptoms including ataxia, myoclonus, excessive daytime drowsiness, headache. After the onset of neurological symptoms, the illness progressed rapidly over the next 2 months with cognitive impairment, particularly memory impairment, myoclonus, ataxia, incontinence of urine and progressive immobility leading to dependency. CSF tests were negative. She was referred to a neurology department where the diagnosis was confirmed by brain biopsy (detailed elsewhere). The patient died in a state of akinetic mutism. DISCUSSION: The clinical features of our patient were consistent with previous descriptions of nvCJD, mainly those of the National CJD Surveillance Unit studies (17): early psychiatric symptoms, prolonged duration of illness (median: 14 months), earlier age at death, compared with sporadic CJD. Psychiatric symptoms occur in the clinical course in about a third of cases of sporadic CJD (3). In contrast, of the 35 cases that have died of nvCJD identified in the study by Will et al. (17), 34 suffered from early and prominent psychiatric symptoms, mainly depression and anxiety. In most of the patients, the first symptoms were psychiatric. Drug treatment was used in most cases, some patients had a transient improvement (18). The patient without psychiatric symptoms reported by the NCJDSU (17) was emotionally labile. Infantile affect and emotional lability, found in our patient, are frequently reported in other studies (1, 18). Schizophreniform disorders have been described during the clinical course, with auditory and visual hallucinations and paranoid delusions (17, 18). The insomnia and excessive daytime drowsiness our patient presented have been described in similar cases (18). Investigations are important to rule out alternative diagnoses. EEG records do not show periodic triphasic complexes as in sporadic CJD. The P 14.3.3 protein in the CSF is positive in half the cases of nvCJD (17). First neurological symptoms developed 6 months after the onset of psychiatric symptoms including ataxia, myoclonus and persistent painful sensory symptoms (17, 18). In most of the cases, MRI brain scans show bilateral pulvinar high signal (17), found subsequently in our patient and detailed elsewhere (13). The terminal stages are progressive cognitive impairment, helplessness and akinetic mutism. CONCLUSION: The first symptoms of this patient were purely psychiatric and difficult to distinguish from common psychiatric disorders. Clinical surveillance of human prion disease is crucial in France, as in UK. The link with BSE has dramatically highlighted the need for neurological and neuropsychological precise investigations. PMID- 11407274 TI - [A case of acute psychotic episode after a single dose of ecstasy]. AB - Over the last 10 years or so in Europe, there has been a development of the "ecstasy" phenomenon, which is the symbol of "recreational" drugs in general. Users, either alone or in private parties, are on the increase. The phenomenon is most frequent in England and in the Netherlands, with an estimated incidence of 13-18% amongst the 18-25 years old, which may reach 52% in "exposed populations", such as individuals who go to "techno" night clubs. In France, the prevalence is uncertain, but estimated at least 5% of males between 18 and 23 years old. Several substance, with more or less the same effects, are grouped together by the term "ecstasy", the best-known one being 3,4-methylenedioxymethamphetamine (MDMA), but there are also an N-demethylated derivative (MDA), methylenedioxyethylamphetamine (MDEA), N-methyl-benzodioxazolylbutanamine (MBDB) and 4-bromo-2,5-dimethoxyphenylethylamine (2-CB or Nexus). The psychopathological consequences of MDMA in man are relatively poorly understood. On the basis of series of cases reported in the literature, acute psychosis, chronic psychosis similar to paranoid delusions, flash-back phenomena similar to with LSD, anxiety/panic states and depressive mood disorders may occur. The case which we report is therefore that of an acute psychotic episode, with residual symptomatology 6 months later, which occurred suddenly following absorption of toxic substances (alcohol and ecstasy), at least 12 hours after taking the ecstasy tablet without his knowing it, in an individual without any previous psychopathology, other than moderate social phobia. Twelve cases of acute psychotic episodes after ecstasy have been reported in the literature. Two of them occurred after a single dose and one after 2 doses. No author was able to examined the previous history of the individuals accurately, nor any possible psychopathological history. Our patient did not have any previous history of psychosis, using a standardized validated interview, which his peers and family confirmed. He did however fulfil the criteria of "social phobia". Retrospectively, we noted the extent of his psychomotor disinhibition with ecstasy, which seemed to be proportional to the intensity of his previous social inhibition. This point does not explain the psychotic episode. From a neurobiologic point of view, acute psychotic disorders are often related to dysfunction of the mesolimbic dopaminergic system. During the first 3 hours, the effect of absorption of MDMA is a massive release of the serotonin, dopamine and noradrenaline stocks. Later, an acute hyposerotoninergic state is present. In our observation, the psychotic disorder appeared at least 12 hours after taking ecstasy, during the reduction phase of the intoxication. Toxicological analysis of the blood was negative (this detection is only positive for 24 hours). Like other authors, our hypothesis is that serotoninergic dysregulation affects the dopaminergic system. Sudden disappearance of serotoninergic feedback on the dopaminergic pathways, may contribute to an increase in the mesolimbic hyperdopaminergic state. In animals, it has been shown that serotonin depletion induced by MDMA, unmasks the effects of a hyperdopaminergic state. In addition, although it has not been mentioned much in the literature, MDMA disturbs dopaminergic function either directly, or through the peptidergic systems (neurotensin, substance P, dynorphines). A consistent series of arguments therefore suggest that there is a sudden central hyperdopaminergic state, which may be related to the appearance, sometimes de novo, of an acute psychotic disorder. From the published cases, psychotic disorders following absorption of ecstasy, appear to become chronic. Most of the cases occurred in individuals, who either abused multiple substances or were chronic ecstasy users. In a case like the one we report, in an individual with good general health, who is not a drug user and who has an acute episode following a single dose, the prognosis should be good. Similarly, a team from Milan has described the experience of 3 friends who had a brief psychotic episode, following ingestion of substances containing ecstasy. These episodes resolved completely, after rehydration and anxiolytic treatment. However, after 6 months' follow-up, our patient still has psychotic symptoms, albeit mild, but which were not present before the intoxication. The patient and his psychiatrist do not envisage changing or stopping his antipsychotic treatment. Other authors have described a lesion in the serotoninergic neurons, by making a parallel with toxic effects described in animals, in particular in primates, with MDMA. Degradation of the serotoninergic cell bodies and nerve endings has been suggested to occur with high doses and/or repeated doses of MDMA. Other authors show the large variations in MDMA and MDA metabolism. (ABSTRACT TRUNCATED) PMID- 11407275 TI - [Diagnostic image (39). Leiomyosarcoma of the lungs]. AB - A 12-year-old girl was unsuccessfully treated for a suspected empyema of the thorax. Afterwards a CT scan, colour doppler flow ultrasonography and a biopsy led to leiomyosarcoma of the lung being diagnosed. PMID- 11407276 TI - [Interactive cd-rom on the choice between breast-sparing treatment and mastectomy: positive responses from patients and surgeons]. AB - OBJECTIVE: To assess how breast cancer patients and surgeons rate an interactive decision aid, using CD-ROM technology, developed to support a choice between breast-conserving therapy and mastectomy. DESIGN: Questionnaire investigation. METHOD: Written questionnaires were presented to patients as soon as they had finished using the CD-ROM. They responded on a 5-point scale. Surgeons (n = 14) were sent a written questionnaire to investigate their experiences of using the CD-ROM program. RESULTS: The interactive CD-ROM was offered to 92 consecutive breast cancer patients. Responses were obtained from 86 (93%) of the patients. The remainder were inadvertently not given the questionnaire. All of the patients evaluated the program as positive. Most of the patients found the program to be 'interesting', 'clear', 'useful', 'encouraging', and 'reassuring'. A majority (n = 76) expressed the view that the amount of information received was 'just right'. The interactive, computer-based method was evaluated as 'agreeable' by 74 patients. Almost all of the patients (n = 82) recommended that the program be shown to other patients. All of the surgeons (n = 14) stated that their patients' evaluations of the program were positive. Moreover, most of the surgeons (n = 10) were of the opinion that the program 'considerably contributed' to the provision of information. With the exception of one surgeon (n = 13), they all held the view that the program can be offered to breast cancer patients facing a choice between breast-conservation or mastectomy on a standard basis. CONCLUSION: Interactive decision-making support by means of a CD-ROM for patients with breast cancer, received positive evaluations from both the patients and surgeons. PMID- 11407277 TI - [Facial paresis in children; consider Lyme disease]. AB - Three girls, aged 3, 7 and 13 years, developed acute peripheral facial palsy. The first patient was initially diagnosed as having Bell's palsy. The third patient had negative serology at first assessment, on the basis of which the diagnosis of Lyme disease was temporarily rejected. Ultimately, all three appeared to have neuroborreliosis. They were treated with intravenous ceftriaxone and recovered well. Facial palsy in childhood is frequently caused by Lyme borreliosis and infection with Borrelia burgdorferi should therefore be investigated, even if there are no signs of a tick bite or erythema migrans. Diagnosis is made by serology, followed by immunoblotting to confirm a positive result. In case of strong suspicion based on the patient's history or physical examination or a positive serology, lumbar puncture should be carried out. Antibiotic treatment facilitates recovery and prevents complications. PMID- 11407278 TI - [Non-specific inflammation of the respiratory tract in a female patient with inflammatory bowel disease and osteoporosis]. AB - A 33-year-old woman presented with a non-specific inflammation of the respiratory passages, which occurred two months after partial colectomy and sigmoidectomy for local stenosis caused by an unclassified inflammatory bowel disease. After other causes of the respiratory symptoms had been ruled out, it was concluded that these were a complication of the bowel disease. Due to the osteoporosis, the patient was given a prolonged treatment with high doses of inhaled corticosteroids instead of systemic corticosteroids. She was treated successfully. PMID- 11407279 TI - [Analysis of expert reports from a general practice expert in disciplinary cases and malpractice claims against family practitioners (1994-2000): a lesson for clinical practice]. AB - Of the reports issued by a general practice expert for disciplinary and malpractice cases in the period 1994 to August 2000, 76 were analysed, with the emphasis on the more serious cases. Infectious diseases and cardiovascular diseases together formed 42% of the cases judged. The starting point of the analysis was the practice of an 'average general practitioner'. On the basis of jurisprudence and general practice literature, failure to diagnose a rare illness was not automatically regarded as reproachable but failing to estimate the seriousness of a situation was. In the first place it was checked whether or not general practitioner had adequately estimated the seriousness of the situation by paying due attention to the alarm signals and risk factors and in the case of diagnostic doubts by ensuring adequate follow-up. According to the analysis, 41% of the cases clearly exhibited reproachable conduct. This figure was twice as high when a locum was working at a practice. Two-thirds of the primary errors were due to the seriousness of the situation not being adequately estimated as a result of too little attention being paid to alarm signals and risk factors as well as a failure to ensure adequate follow-up. On the basis of the analysis it is advised that the use of a locum should be regarded as a risk situation: better information can be made available by means of an electronic patient file and unequivocal agreements for drawing up reports. For each consultation, the GP must systematically check whether there are alarm signals or risk factors. In the case of diagnostic doubts the clinical picture should be actively followed up by means of clear follow-up appointments at definite times. In the guidelines of the Netherlands Society for General Practitioners, explicit attention should be given to alarm signals and risk factors. In medical training and ongoing professional education, systematic training should be provided in the recognition of serious situations and rare diseases/complications. PMID- 11407280 TI - [Loss of an eye due to hyper-IgE syndrome after a corneal transplant]. PMID- 11407281 TI - [Lambing sheep as source of severe psittacosis in pregnancy]. PMID- 11407282 TI - [Surgery for a recently symptomatic severe carotid stenosis or not? Clinical application of 'evidence-based' medicine]. PMID- 11407283 TI - [Neostigmine for the treatment of acute pseudo-obstruction of the colon (Ogilvie syndrome)]. PMID- 11407284 TI - [Boerhaave's syndrome: also in the emergency room]. PMID- 11407285 TI - [Erectile dysfunction: prevalence and effects on the quality of life: the Boxmeer study]. PMID- 11407286 TI - [An eye on the vocal cords: family practitioner and hoarseness]. AB - In four patients with hoarseness, men aged 53 and 67 years, and women aged 8 and 37 years, indirect laryngoscopy revealed, respectively, a squamous cell carcinoma of the vocal cord, recurrent nerve paralysis due to pulmonary carcinoma, irritative noduli due to forced use of the voice, and psychic stress as the cause. In a patient with existing hoarseness over a period of 3 to 6 weeks, the vocal cords will have to be examined with indirect laryngoscopy. Where the general practitioner is not equipped to handle this procedure, he can make a referral to an ear, nose and throat specialist. It would, however, be to the benefit of the patient and the general practitioner if the latter were to master the technique of indirect laryngoscopy. This would enable the patient to be treated without further delay, and it might also make selective referral possible. PMID- 11407288 TI - What's the matter of depth of anesthesia? PMID- 11407287 TI - [CBO guideline 'larynx carcinoma']. AB - The guideline 'Larynx carcinoma' aims at preventing divergent views on the treatment of different stages of larynx carcinoma between the major referral centres. Strict criteria are proposed for the radiological diagnostic procedures and the pathology report in order to accurately classify the patient in the appropriate prognostic category. Taking into consideration possible subsequent treatments, surgery--preferably by laser--is proposed for severe dysplasia and carcinoma in situ. For all other stages the treatment of choice is radiotherapy as a larynx conserving therapy. Only in cases of massive tumour volumes with invasion through the laryngeal skeleton where the chance of cure is low with radiotherapy, surgery is the treatment of choice. Minimal criteria are described for rehabilitation and supportive care. Due to the relative rarity of the disease and the complexity of the treatment, all patients with a laryngeal carcinoma should be seen at least once in a dedicated head and neck clinic. PMID- 11407289 TI - EEG-bispectral index changes with ketamine versus thiamylal induction of anesthesia. AB - BACKGROUND: The EEG-Bispectral Index (BIS) is a processed EEG information that has been validated as a means to measure the hypnotic effect of anesthetic drugs. In this study we evaluated the BIS changes in induction of anesthesia with ketamine in comparison with that of thiamylal. METHODS: Forty ASA class I and II adult female patients undergoing elective gynecologic surgeries were enrolled into this randomized, prospective study. No premedication was given to the patient. In each patient EEG was recorded continuously from the frontal electrodes using Aspect A-1050 monitor after his arrival at the operating room. Blood pressure and heart rate were also recorded throughout the surgery. After steady baseline recordings of all necessary parameters having been accomplished Group K patients (n = 20) were given an induction dose of ketamine 1.5 mg/kg i.v., whereas Group T patients (n = 20) received thiamylal 5 mg/kg i.v. When loss of consciousness was ascertained, intubation was performed after administration of succinylcholine 1 mg/kg i.v. and anesthesia was maintained with isoflurane nitrous oxide-oxygen. Demographics, BIS values, HR, BP were analyzed and compared. RESULTS: The demographics were comparable between the two groups. Both groups showed a mean value of BIS of 96 with comparable BP and HR before induction. After study drug the post-induction BIS for ketamine was 94 as against 51 for thiamylal (P < 0.05), 91 against 43 post-succinylcholine (P < 0.05), 92 against 53 post-intubation (P < 0.05) and 45 against 37 five min after isoflurane. BIS remained below 60 throughout the entire course of anesthesia and returned to above 95 on emergence in both groups. None of the patients reported awareness, recall, delirium or hallucination during anesthesia. CONCLUSIONS: Ketamine-induced dissociative anesthesia produces persistently elevated BIS index which is different from thiamylal and those reported with other conventional anesthetic agents. The established range of BIS index appears not to be applicable in patients under ketamine anesthesia. Monitoring the depth of ketamine anesthesia remains to be a challenging problem. PMID- 11407290 TI - Effects on the bispectral index during elective caesarean section: a comparison of propofol and isoflurane. AB - BACKGROUND: Awareness during general anesthesia has been a particular problem during caesarean section. About 7 percent of patients undergoing elective caesarean section have reported dreaming or recall of voices during the procedure. The bispectral index (BIS), a value derived from the electroencephalogram (EEG), has been shown to be useful in monitoring the depth of anesthesia. Supplementation of propofol or isoflurane for maintenance of anesthesia has been shown to effectively reduce the incidence of awareness. However, the effects of propofol or isoflurane on the BIS index have not been fully investigated. We therefore designed this study to compare the effects of isoflurane or propofol supplementation on the BIS index in 24 healthy parturients undergoing elective caesarean section. METHODS: All patients had induction of anesthesia and orotracheal intubation in rapid sequence made possible by 1 MAC isoflurane with 50% N2O-50% O2 as conveyer and atracurium. After delivery, patients were randomly assigned to either of two groups (isoflurane or propofol). Patients in the Isoflurane group (n = 12) received 0.5 MAC isoflurane in 67% N2O 33% O2 and fentanyl + droperidol. Patients in the propofol group (n = 12) received propofol (8 mg/kg/h) infusion combined with 67% N2O-33% O2 and fentanyl + droperidol. RESULTS: There was no difference between the two groups in total operation time, maternal blood loss, fetal Apgar scores. No differences between the two groups in heart rate, blood pressure or BIS index values were found throughout the surgery. No patient from either group reported recall of the operative procedure. However, 25% of patients (3 of 12) in the isoflurane group had poor uterine contraction, suggestive of doubtful appropriateness of the use of isoflurane for maintenance of anesthesia in delivery. CONCLUSIONS: We therefore concluded that supplementation of isoflurane or propofol for maintenance of anesthesia can satisfactorily decrease the BIS index and minimize the incidence of awareness in patients undergoing caesarean section under general anesthesia. The BIS index is a reliable monitor of the hypnotic component of anesthesia. PMID- 11407291 TI - Rapid recovery of spontaneous baroreflex after sevoflurane anesthesia in ambulatory surgery. AB - BACKGROUND: Although sevoflurane seems to have fulfilled the criteria of ideal anesthetic agent for ambulatory surgery, its effects on intraoperative alteration and postoperative recovery of arterial baroreflex have not been well documented. This study assessed the time required for patients anesthetized with sevoflurane to regain their baseline baroreflex after ambulatory surgeries. METHODS: Ten ASA class I female patients scheduled for minor gynecological operation (D&C) were enrolled in this study. Spontaneous baroreflex sensitivity (SBR) calculated by sequence analysis of beat-to-beat variations in systolic arterial pressure and R R intervals was recorded before anesthesia (Baseline), during anesthesia (Anesthesia), five min after operation (Post-op 1) and ten min after operation (Post-op 2). Data obtained from Anesthesia, Post-op 1, and Post-op 2 was compared with the Baseline. RESULTS: SBR value determined in Baseline group (28.6 +/- 4.9 ms/mmHg) was significantly different from Anesthesia group (12.2 +/- 2.1 ms/mmHg) and Post-op 1 group (15.8 +/- 1.8 ms/mmHg). Patients regained their conscious baseline baroreflex sensitivity within ten min after sevoflurane was switched off. CONCLUSIONS: Sevoflurane anesthesia depresses SBR and provides a quick SBR recovery upon emergence. PMID- 11407292 TI - Prolonged interference of patent blue on pulse oximetry readings. AB - BACKGROUND: We evaluated the effect of intra-arterial injection of Patent Blue (PB) on pulse oximetry. METHODS: Ten consecutive female patients who underwent intra-arterial insertion of catheter for chemotherapy were enrolled in this study. All patients received general anesthesia and ECG, pulse oximetry, and direct arterial blood pressure were monitored throughout the perioperative period. Baseline arterial blood saturation (SpO2), haemoglobin concentration (Hgb), estimated arterial saturation (SaO2), arterial oxygen partial pressure (PaO2) were obtained through arterial blood gas analysis, and true arterial saturation (ScoO2) through CO-Oximetry analysis. The subsequent values obtained at various points of time following the intra-arterial administration of PB were compared with the baseline data. RESULTS: The SpO2 readings taken after intra arterial injection of PB were significantly lower than the baseline reading. The extent of the reduction of SpO2 readings two hours after the administration of PB was relevant to the total accumulative dosage of PB (rs = -0.846, P < 0.05). However, the reduction of SpO2 bore no significant relation with Hgb concentration or with the ratio of PB dosage to Hgb concentration. CONCLUSIONS: Intra-arterial administration of PB interferes with pulse oximetry readings and the effect lasts for a variable duration. The extent of desaturation may be related to the total accumulative dosage of intra-arterial PB injected. PMID- 11407293 TI - Effectiveness and safety of rocuronium-hypnotic sequence for rapid-sequence induction. AB - BACKGROUND: Either succinylcholine or rocuronium administered after a hypnotic is the current technique for rapid-sequence induction. It is assumed that rocuronium administered before a hypnotic (Rocuronium-hypnotic sequence) may equally provide an acceptable intubation condition as well as a shorter period of apnea in rapid sequence induction. We designed a prospective, randomized study to evaluate the effectiveness and safety of the technique in a similar rapid-sequence induction. METHODS: Ninety adult patients receiving elective surgeries were enrolled in this study. In all patients the procedure in the study began with i.v. injection of fentanyl 2 micrograms/kg, followed by preoxygenation with 100% O2 for 2 min. Afterward, the patients were randomly allocated to 3 groups with each group consisting of 30 patients. In Rocuronium-thiopental (Ro-Th) group the patients received rocuronium 0.6 mg/kg and then thiopental 5 mg/kg; in Th-Ro group the patients received thiopental 5 mg/kg and then rocuronium 0.6 mg/kg; and in Thiopental-Succinylcholine (Th-Sx) group, the control group, the patients received thiopental 5 mg/kg and then succinylcholine 1 mg/kg. Laryngoscopy and endotracheal intubation were performed 60 s after the injection of the muscle relaxant. The intubation condition, the apneal time before laryngoscopy, the intubation time, and total apneal time were investigated and compared. Presence of injection pain, sense of paralysis, SpO2 less than 95% during induction, and any unexpected adverse event were also recorded. RESULTS: Six patients (1 in Ro Th group, 2 in Th-Ro group, and 3 in Th-Sx group, respectively) were excluded from the study. The intubation conditions were acceptable in all patients of three groups who completed the study, and as to excellent intubation condition there was no difference between the three groups. In Ro-Th group both the apneal time before laryngoscopy (32.4 +/- 5.4 s) and total apneal time (48.5 +/- 11.0 s) were the shortest. Th-Ro group (53.2 +/- 5.8 and 67.5 +/- 8.3 s, respectively) and Th-Sx group (54.4 +/- 5.8 and 68.4 +/- 7.7 s, respectively) were similar in both aspects. With respect to intubation time there was no significant difference among the three groups. Five patients in Ro-Th group and one patient in Th-Sx group felt mild injection pain. Three patients in Ro-Th group were noted to have diminished breathing during induction, which was not recalled during enquiry in the postoperative visit. One patient in Ro-Th group saw a fall of SpO2 down below 95% (94% the minimal) during the apnea period. CONCLUSIONS: Compared with traditional hypnotic-rocuronium or hypnotic-succinylcholine sequence, rocuronium (0.6 mg)-thiopental sequence can provide a similar intubation condition but cause a much shorter apneal period in rapid-sequence induction. In carrying out recuronium-thiopental sequence induction, maintaining a patent infusion line is essential to avoid drug precipitation and awareness of muscular weakness as a result of ill-timed action of thiopental. PMID- 11407294 TI - A preliminary report on the Chinese Cancer Pain Assessment Tool (CCPAT): reliability and validity. AB - BACKGROUND: This study was to establish the validity and reliability of the Chinese Cancer Pain Assessment Tool (CCPAT) for Chinese patients with cancer pain in Hong Kong. METHODS: It was a quasi-experimental study. The CCPAT was tested for its reliability and validity by 26 cancer pain patients and 26 chronic non cancer pain patients. McGill Pain Questionnaire (MPQ), with its established reliability and validity, was used as the criterion tool for comparison. RESULTS: An internal consistency of 0.88 and the inter-rater reliability of 0.96 were obtained for the CCPAT. The Spearman's Rho correlations between the functional and emotional dimensions of the CCPAT and the present pain intensity of the MPQ positively correlated. This indicated a satisfactory concurrent validity. Discriminant validity was performed with MANOVA. The CCPAT was able to predict 80.8% of the subjects correctly as cancer pain patient from the cancer pain group. CONCLUSIONS: The constructed CCPAT is a valid and reliable cancer pain assessment tool in a Chinese context. The developed CCPAT is the first of its kind for the Chinese community in Hong Kong. It is anticipated that the results can provide the health care professionals with the best possible instrument and indicator. The CCPAT also lays the theoretical construct for the professionals with a better understanding of the experience of cancer pain from a multidimensional perspective. PMID- 11407295 TI - Repeated attacks of venous air embolism during craniotomy--a case report. AB - Venous air embolism (VAE) is not uncommon during craniotomy, but repeated attacks of VAE during a single surgical procedure is rarely seen. We report a successful intraoperative management of repeated attacks of air embolism in a patient who sustained craniotomy for intracranial hemorrhage (ICH) in prone position. A 70 year-old male suffering from hemorrhage in the right cerebellar hemisphere with impending brainstem herniation was scheduled for craniotomy. He had history of hypertension but it was not well controlled with medical treatment. Emergent craniotomy for removal of blood clot resulting from ICH was performed. During the operation, sudden decrease of end-tidal CO2 (EtCO2) level, fall of blood pressure and increase of central venous pressure (CVP) were noted. Since air bubbles were retrieved from CVP catheter venous air embolism was highly suspected. With prompt diagnosis and proper management, we successfully improved the patient's hemodynamic status and he was discharged without any sequelae. Early detection together with aggressive treatment is the only way in the management of intraoperative venous air embolism. PMID- 11407296 TI - Peripartum cardiomyopathy--a case report. AB - Peripartum cardiomyopathy in the form of congestive heart disease of unknown etiology appears relatively rarely during the last month of pregnancy or within 5 months after delivery, and it is potentially life-threatening. This case report describes a 34-year-old female, G4P1, at 36 weeks' gestation who was admitted for cesarean section under lumbar epidural anesthesia due to twin pregnancy. She had no past history of cardiovascular diseases. However, rapid onset of dyspnea at ward and acute cardiac failure developed 15 h after cesarean section. Peripartum cardiomyopathy was diagnosed. After a 7-day intensive treatment she was discharged. Since then she was symptom-free and her two babies were doing well during a period of one more years after discharge. PMID- 11407297 TI - Endotracheal tube fires during carbon dioxide laser surgery on the larynx--a case report. AB - Endotracheal tube (ETT) fire is a catastrophic disaster that may occur during laser surgery of the upper airway. Several means are available for protection of polyvinyl chloride (PVC) tube from fire, but they are not perfect in prevention of fires caused by laser beam. The PVC tube is hazardous for carbon dioxide (CO2) laser surgery if it is not well wrapped with metallized foil tape. We report a case that a PVC ETT wrapped with aluminum foil ignited during CO2 laser surgery of the larynx. In this report, we emphasize the shaft of the PVC tube must be completely wrapped with aluminum foil to prevent exposure of any surface if it is used in CO2 laser surgery of the upper aero digestive tract. PMID- 11407298 TI - Gender differences in autoimmunity: molecular basis for estrogen effects in systemic lupus erythematosus. AB - Systemic lupus erythematosus (SLE) is an autoimmune disease that occurs primarily in women (9:1 compared to men). Estrogen is a female sex hormone that acts on target cells through specific receptor proteins and alters the rate of transcription of target genes. Experiments in our laboratory have shown that calcineurin steady-state mRNA levels and phosphatase activity increase when estrogen is cultured with SLE T cells. This estrogen-dependent increase is dose dependent, hormone-specific and temporally regulated. Estrogen receptor antagonism by ICI 182,780 inhibits the increase in calcineurin mRNA and phosphatase activity, while cycloheximide has no effect suggesting that new protein synthesis is not required. Reverse transcription and polymerase chain amplification indicate that estrogen receptor-alpha and estrogen-beta are expressed in human T cells. However, calcineurin does not respond to estrogen stimulation in T cells from normal females, males and lupus males. Taken together, these results indicate a differential function of the estrogen receptor in women with lupus. A model is proposed that suggests estrogen, acting through the estrogen receptor, enhances T cell activation in women with lupus resulting in amplified T-B cells interactions, B cell activation and autoantibody production. PMID- 11407299 TI - Estrogen and progesterone receptors in murine models of systemic lupus erythematosus. AB - Estrogens are believed to play a role in the etiology of both human and murine systemic lupus erythematosus (lupus; SLE), presumably through the agency of their cellular receptor proteins. There is now considerable interest in the molecular mechanism of action of estrogens in immune tissues, particularly with regard to autoimmune disorders, which are generally more prevalent in women. In this laboratory, an attempt is being made to characterize estrogen receptors in murine models of SLE and to try and relate this to estrogen receptor function in vivo. The initial aim was to compare binding properties of estrogen receptors in brain, reproductive and immune tissues of BALB/c and MRL/MP-lpr/lpr mice. The latter strain spontaneously develops an autoimmune disease resembling human systemic lupus erythematosus (lupus; SLE). It is hypothesized that estradiol, through its receptors, mediates the progression of murine SLE, and that in autoimmune disease, the estrogen receptor is functionally and/or structurally changed. Initial studies suggest that there are differences in estrogen receptors between BALB/c mice, which do not get autoimmune disease, and two strains that do, MRL/MP lpr/lpr and NZB/W mice. In MRL mice, these differences may be reflected in impaired priming of the progesterone receptor. PMID- 11407300 TI - Progesterone as an immunomodulatory molecule. AB - Increased progesterone sensitivity of pregnancy lymphocytes is due to activation induced appearance of progesterone binding sites in the lymphocytes. Following recognition of fetally derived antigens gamma/delta TCR+ cells develop progesterone receptors. Progesterone binding results in the synthesis of a mediator protein named the progesterone-induced blocking factor (PIBF). PIBF by acting on the phospholipase A2 enzyme interferes with arachidonic acid metabolism, induces a Th2 biased immune response, and by controlling NK activity exerts an anti-abortive effect. PMID- 11407301 TI - Membrane estrogen and glucocorticoid receptors--implications for hormonal control of immune function and autoimmunity. AB - Membrane steroid receptors (mSRs) have recently re-emerged as candidates for mediating steroid effects which do not fit the paradigm of nuclear transcription factor mechanisms. We have studied two steroid-binding classes of mSRs, and have noted striking similarities in their characteristics (immunocytochemical appearance, biochemical properties, proteolytic sensitivity, signaling pathways, regulation, and molecular origins). These observations strengthen the conclusion that mSRs can be modified versions of intracellular steroid receptors. The membrane estrogen receptors (mERs) we studied are involved in estrogen-induced release of prolactin. Membrane glucocorticoid receptors (mGRs) in both mouse and human lymphoma cells are necessary for the initiation of glucocorticoid-induced therapeutic apoptosis which is related to the developmental phenomenon of thymic involution. Diseases of autoimmunity such as systemic lupus erythematosus and arthritis are related to estrogen status. Since both of these mSRs have recently been found in both normal and cancerous lymphoid cells, actions of these mSRs may have important consequences for functions and diseases of the immune system. Therefore, the study of these forms of steroid receptors may present novel therapeutic opportunities for the use of steroids and steroid analogs. PMID- 11407302 TI - Treatment of systemic lupus erythematosus. AB - The treatment of systemic lupus erythematosus (SLE) is mainly based on a number of "traditional" drugs such as corticosteroids, antimalarials, azathioprine and cyclophosphamide. However, this scenario is rapidly changing due to the introduction of new compounds. Some of these new agents have been successfully used in other diseases, while others are being specifically designed to interfere with the immune abnormalities seen in SLE. As our knowledge on the mechanisms of immune response increases, new drugs that can interfere with T and B cell interaction and activation, production of anti-dsDNA autoantibodies, immune complexes deposition and cytokine activation have been developed and some of these are now under investigation in SLE. Although initial data regarding their safety and efficacy are encouraging, caution must be taken before these drugs are considered as the treatment of choice for specific SLE manifestations. Specifically, controlled clinical trials with sufficient number of patients are necessary. If the promising results already available are confirmed, the use of these drugs might represent the keystone in the future management of SLE and other autoimmune diseases. PMID- 11407303 TI - Gonadotropin-releasing hormone: potential role in autoimmunity. PMID- 11407304 TI - Immunomodulatory effects of interferon-beta-1b in patients with multiple sclerosis. AB - The mechanisms by which IFN beta-1b acts in the treatment of patients with multiple sclerosis (MS) are not completely known. Immunomodulatory effects of IFN beta-1b were investigated in patients with relapsing-remitting (RR) MS in vivo and in vitro. Compared to baseline and controls, defined as patients with RR-MS without immunomodulatory therapy, the expression of TGF beta-1-mRNA by peripheral blood mononuclear cells (PBMC) was persistently increased at week 6, month 3 and month 6 (p < or = 0.05), that of the TGF beta-1 receptor type II from day 5 up to month 6 (p < 0.01). The expression of TNF alpha-mRNA decreased from day 1 to month 3 compared to day 0 and the controls (p < 0.01). The in vitro investigations performed on isolated peripheral blood lymphocytes demonstrated that these effects were dose-dependent. The mRNA and protein expression of TNF alpha-R-I (55 kD-receptor) was only temporarily elevated at the beginning of the therapy in vivo. The expression of TNF alpha-R-I-mRNA increased dose-dependently after stimulation with IFN beta-1b for 24 h in vitro. Serum levels of soluble vascular cell adhesion molecule (sVCAM) were increased during the whole time of in vivo treatment (p < 0.01). The CD8CD38 lymphocyte subpopulation was continuously elevated from day 5 up to month 6 (p < 0.01) in the MS patients treated with IFN beta-1b in vivo. No persistent, significant changes were demonstrable concerning the percentage of total CD4, CD8, CD19 nor in CD4 subpopulations (CD4CD29, CD4CD45RA). The present data suggest that IFN beta-1b induces the mRNA expression of TGF beta-1 and TGF beta-R-II by PBMC, decreases that of TNF alpha and increases levels of sVCAM-1 and of circulating activated CD8 cells (CD8CD38) in blood. These might be other mechanisms by which IFN beta 1b mediates its positive effects in the treatment of MS patients. PMID- 11407305 TI - A role for parathyroid hormone-related protein in the pathogenesis of inflammatory/autoimmune diseases. AB - Our increased understanding of the critical role of cytokines in chronic inflammatory/autoimmune diseases has led to the recent development of effective anti-cytokine treatments. In particular, agents blocking the function of TNF alpha, a cytokine first identified as an endotoxin-inducible mediator of tumor cell necrosis, are now licensed for the treatment of rheumatoid arthritis (RA) and inflammatory bowel disease. However, TNF-alpha is but one member of a cytokine network that is responsible for mediating these inflammatory disorders. Therefore, as our understanding of the pathophysiologic role of other members of this inflammatory network increases, other cytokines may similarly be identified as effective targets for treatment. In this article, we will review evidence which suggests that parathyroid hormone-related protein (PTHrP), a peptide which, like TNF-alpha, was first identified because of its effects in the setting of malignancy, may in fact serve an important non-neoplastic, physiologic function by mediating the inflammatory/autoimmune host response. Data identifying PTHrP as a member of the cytokine network induced in multi-organ inflammation and rheumatoid arthritis will be summarized, initial evidence comparing the therapeutic efficacy of PTHrP- vs. TNF-alpha-blockade in the treatment of endotoxemia will be reviewed, and potential future areas of research, including assessment of the effects of PTHrP blockade in the treatment of RA, will be discussed. PMID- 11407306 TI - Effects of Linomide on immune cells and cytokines inhibit autoimmune pathologies of the central and peripheral nervous system. AB - Linomide (roquinimex, LS 2616) is a quinoline-3-carboxamide with pleiotropic immune modulating capacity and it has therapeutic effects in several experimental animal models of autoimmune diseases. Linomide has been evaluated in clinical trials for multiple sclerosis, and was indeed shown to have disease inhibitory effects. However, due to unexpected side effects recorded in patients treated with Linomide, premature termination of clinical trials was required. The basic mechanism(s) of action of Linomide in inducing beneficial effects in autoimmune diseases is still elusive. Some experimental evidence indicates that Linomide influences the regulation of the cytokine profile, resulting in the inhibition of autoimmune and inflammation pathologies. This review focuses on Linomide applied in models for autoimmune and inflammation pathologies of the central and the peripheral nervous system, and summarises its very encouraging disease inhibitory effects and their potential pharmacological basis. The beneficial effects recorded with Linomide in both experimental and clinical trials emphasise the possible value of substances with Linomide-like activity for clinical use in autoimmune and inflammation pathologies in the near future. PMID- 11407307 TI - The immunomodulator Linomide: role in treatment and prevention of autoimmune diabetes mellitus. AB - Insulin-dependent diabetes mellitus (IDDM) is considered to be an autoimmune disorder characterized by destruction of the pancreatic beta-cells by auto reacting lymphocytes. An attractive therapeutic approach to this disease would be to abrogate the autoimmune process at an early stage, thus preserving a critical mass of pancreatic beta-cells necessary for maintenance of normal glucose tolerance. Linomide (quinoline-3-carboxamide, Roquinimex, LS 2616), is a novel, orally absorbed, immunomodulatory drug that has been shown to be effective in various models of autoimmunity without causing non-specific immunosuppression. In this review, we describe the efficacy of Linomide for ameliorating the autoimmune process and diabetes in the non-obese diabetic (NOD) model of IDDM when administered at early stages of the disease. We also show that advanced disease in the NOD mouse can be treated effectively by combining Linomide with therapeutic modalities designed to increase pancreatic beta-cell mass. Subsequent clinical studies have shown that Linomide preserves beta-cell function in individuals with new-onset IDDM. Based on these data, Linomide or derivatives thereof might be useful for treatment of human IDDM. PMID- 11407308 TI - Aqueous extract isolated from Platycodon grandiflorum elicits the release of nitric oxide and tumor necrosis factor-alpha from murine macrophages. AB - Herbal medicines are increasingly being utilized to treat a wide variety of disease processes. Aqueous extract from the root of Platycodon grandiflorum A. DC (Campanulaceae), Changkil (CK), is reported to have antitumor and immunomodulatory activities; however, the mechanism underlying its therapeutic effect is not known. In the present study we examined the effects of CK on the release of nitric oxide (NO) and tumor necrosis factor-alpha (TNF-alpha), and on the gene expression of iNOS and TNF-alpha in mouse macrophages. CK elicited a dose-dependent increase in NO and TNF-alpha production in cultured macrophages. CK significantly affected secretion at concentrations of more than 5 micrograms/ml, and its maximum effect was at concentration of 100 micrograms/ml. Reverse transcription polymerase chain reaction showed that increases in NO and TNF-alpha secretion were due to an increase in inducible NO synthase mRNA and TNF alpha mRNA, respectively. Transient expression assays with NF-kappa B binding sites linked to the luciferase gene revealed that CK-induced increase of inducible NO synthase mRNA and TNF-alpha mRNA were mediated by the NF-kappa B transcription factor complex. These results demonstrate that CK stimulates NO and TNF-alpha release and is able to upregulate iNOS and TNF-alpha expression through NF-kappa B transactivation and this may be a mechanism whereby this herbal medicine elicits its therapeutic effects. PMID- 11407309 TI - Induction of CD4+ regulatory T cells by 12-O-tetradecanoylphorbol 13-acetate in mice. Part 1: The strain difference in this induction. AB - 12-O-Tetradecanoylphorbol 13-acetate (TPA), an activator of signal transduction, induced antigen-nonspecific regulatory T (Tr) cells for delayed-type hypersensitivity (DTH) in the spleen. A marked strain difference in the induction of Tr cells was observed when the study was performed by using the several strains of mice at 6-8 weeks old. TPA painting induced CD4+Tr cells in C3H/He (H 2k), C3H/HeN (H-2k), DBA/2 (H-2d) and AKR/N (H-2k) mice, but not in C57BL/6 (H 2b), C57BL/10 (H-2b), BALB/c (H-2d) and A/J (H-2a). Regulatory cells were also induced by incubating spleen cells from unprimed mice with TPA in vitro and were seemed to act by the production of soluble factor(s). A downregulatory activity of the soluble factor(s) was abrogated by SXC-1 (anti-IL-10 monoclonal antibody), but not by SXC-2 (anti-IL-10 monoclonal antibody) and anti-IL-4. For purification of the factor(s), we established the T cell hybridoma 4C5-1 by the fusion of spleen cells from TPA-treated C3H/He mice with AKR thymoma Bw5147 cells. The 4C5 1 cells secrete the factor(s) which can inhibit DTH response. The inhibitory activity of the factor(s) could be neutralized by SXC-1, but not by SXC-2, anti IL-4, anti-IL-6 and anti-TGF-beta antibodies. The factor(s) could not affect the proliferation and IFN-gamma production of alpha s1-casein-specific 3D20 Th1 cells. The factor(s) termed DIF (DTH Inhibitory Factor) may be a novel cytokine, since they have reduced the footpad swelling response by local injection, and have no immune crossreactivity with the DTH regulatory cytokines and no inhibitory activity for in vitro Th1 response. PMID- 11407310 TI - Mechanism of action of disease modifying anti-rheumatic agent, gold sodium thiomalate (GSTM). AB - GSTM has been used for long in the treatment of rheumatoid arthritis (RA). However, its mechanism of action is still poorly understood. In the last decade, tumor necrosis factor-alpha (TNF-alpha) has emerged as the major pro-inflammatory cytokine in the pathogenesis of RA. We studied the effect of GSTM on spontaneous and LPS-stimulated TNF-alpha production by human peripheral blood mononuclear cells (PBMCs) of normal volunteers. PBMCs were isolated from 20 normal volunteers and cultured in the presence of absence of lipopolysaccharide (LPS 10 ng/ml) and GSTM (1 microgram/ml). TNF-alpha level was measured using commercial enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). The TNF-alpha response to LPS was heterogeneous. PBMCs of 24 subjects showed high LPS-stimulated TNF-alpha production (LPS-responsive group), whereas that of six individuals had low LPS-stimulated TNF-alpha production (LPS non-responsive group). GSTM-stimulated spontaneous TNF-alpha production and inhibited LPS-stimulated TNF-alpha production in 16 of 24 (75%) individuals of LPS-responsive group and one of six individuals (17%) of LPS-non-responsive group. The suppression of TNF-alpha by GSTM was also demonstrated at the mRNA level. We conclude that there is a heterogeneity among normal population for TNF alpha production in response to LPS, and GSTM inhibits LPS-stimulated TNF-alpha production, primarily in LPS responders. Further study is needed to establish the relationship between LPS responsiveness and GSTM suppression. PMID- 11407311 TI - Decreased serum IgE level, decreased IFN-gamma and IL-5 but increased IL-10 production, and suppressed cyclooxygenase 2 mRNA expression in patients with perennial allergic rhinitis after treatment with a new mixed formula of Chinese herbs. AB - A new mixed formula of Chinese herbs containing Shin-yi-san + Xiao-qing-long-tang + Xiang-sha-liu-jun-zi-tang by the weight of 9 + 3 + 3 g/day was prescribed for the treatment of patients with perennial allergic rhinitis for 3 months (the composition of each herb is shown in the tables of the article). We classified the patients into high (H-IgE) and low IgE (L-IgE) groups according to the titer of serum total IgE (> 200 KIU/l in H-IgE vs. < 200 KIU/l in L-IgE) and the presence of house dust mite-specific IgE. The nasal symptomatic score in the high IgE group was significantly improved from 7.19 +/- 0.18 before treatment to 2.67 +/- 0.37 after treatment. In addition, the serum total and house dust mite specific IgE level were also decreased after treatment. For elucidating the working mechanism of the mixed formula, the Th1 (IFN-gamma) and Th2 (IL-4, IL-5, IL-10 and IL-13) cytokine production by phytohemagglutinin (PHA)-stimulated mononuclear cells (2 x 10(6) cells/ml) and cyclooxygenase type 2 (COX-2) mRNA expression in LPS or IL-13-stimulated PMN were compared before and after 3 months of treatment. We found that the mixed formula treatment significantly enhanced IL 10 but decreased IFN-gamma and IL-5 production by PHA-stimulated mononuclear cells. The IL-5 production was also decreased by PHA-stimulated lymphocyte. In addition, the COX-2 mRNA expression in stimulated PMN was significantly suppressed after treatment. These results suggest that the new mixed formula treatment is beneficial to the patients with perennial allergic rhinitis via modulating the function of lymphocytes and neutrophils. PMID- 11407312 TI - Effects of suplatast tosilate (IPD Capsules) on the production of active oxygen by neutrophils and of IL-8 by mononuclear cells. AB - In bronchial asthma, eosinophils and neutrophils are activated, so that the production of active oxygen species increases, causing airway epithelial injury. Suplatast tosilate (IPD Capsules) is a novel immunomodulating antiallergic drug that acts against bronchial asthma through a new mechanism. To evaluate the effects of suplatast tosilate on mononuclear cell-mediated IL-8 production, and neutrophil-mediated active oxygen species production at sites of inflammation, we collected peripheral blood from healthy subjects and separated the neutrophils as well as mononuclear cells. Suplatast tosilate was added at a concentration of 1 x 10(-6), 1 x 10(-7) or 1 x 10(-8) M, and cells were incubated for 10 min at 37 degrees C. Then, the neutrophils were stimulated with fMLP, and luminol-dependent chemiluminescence (LDCL) was measured, while IL-8 production was determined with an ELISA kit. Suplatast tosilate (1 x 10(-6) M) inhibited neutrophil-mediated active oxygen species production by 12.4% in terms of the peak, and by 16% in terms of the integral value. Moreover, it significantly inhibited mononuclear cell-mediated IL-8 production at concentrations of 1 x 10(-6), 1 x 10(-7) and 1 x 10(-8) M, in a concentration-dependent manner. This study indicated that suplatast tosilate may inhibit neutrophil infiltration by suppressing monocyte mediated IL-8 production, and it may also inhibit the activation of neutrophils at sites of inflammation. These results suggest the possibility that suplatast tosilate may not only be of benefit for asthma, but may also prevent or control pulmonary fibrosis or emphysema, for which no effective treatment is presently available. PMID- 11407313 TI - A catalytic antibody against cocaine attenuates cocaine's cardiovascular effects in mice: a dose and time course analysis. AB - The murine monoclonal antibody 15A10 (mAb 15A10), elicited by a transition-state analog for cocaine hydrolysis, has previously been shown to metabolize cocaine in vitro and in vivo. The present experiments were designed to evaluate further the in vivo effectiveness of mAb 15A10 in blocking cardiovascular effects of acute cocaine administration. Balb/c mice were implanted with a femoral artery catheter utilized for mean arterial pressure (MAP) monitoring, and administered intravenous (i.v.) pretreatments of either mAb 15A10 (10, 32, 100 and 300 mg/kg) or vehicle prior to cocaine injection (100 mg/kg, i.p.). A time course analysis for mAb 15A10's effect was also conducted, for which either vehicle or 100 mg/kg mAb 15A10 was infused 1, 3, 10 and 30 days prior to cocaine treatment. During the cardiovascular recording sessions, mice were awake and freely moving within a limited area. Increases in MAP (approximately 25 mm Hg) following cocaine injection were dose-dependently attenuated by mAb 15A10. The antibody-attenuated cocaine-induced increases in MAP at 1- and 3-day pretreatment times, and reduced mortality at some of the time points studied. With 100 mg/kg antibody, plasma cocaine levels were significantly decreased early in the recording session, whereas levels of ecgonine methyl ester increased significantly. Although 10-fold greater quantities of antibody are required to observe significant effects in mouse, compared to our previous studies in rats, the present mouse model provides a convenient paradigm for investigating catalytic and non-catalytic antibodies. PMID- 11407314 TI - Purine nucleoside phosphorylase inhibitor BCX-1777 (Immucillin-H)--a novel potent and orally active immunosuppressive agent. AB - Patients with purine nucleoside phosphorylase (PNP) deficiency present a selective T-cell immunodeficiency. Inhibitors of PNP are, therefore, of interest as potential T-cell selective immunosuppressive agents. BCX-1777 is a potent inhibitor of PNP from various species including human, mouse, rat, monkey and dog, with IC50 values ranging from 0.48 to 1.57 nM. BCX-1777, in the presence of 2'-deoxyguanosine (dGuo, 3-10 microM), inhibits human lymphocyte proliferation activated by various agents such as interleukin-2 (IL-2), mixed lymphocyte reaction (MLR) and phytohemagglutinin (PHA) (IC50 values < 0.1-0.38 microM). BCX 1777 is a 10-100-fold more potent inhibitor of human lymphocyte proliferation than other known PNP inhibitors like PD141955 and BCX-34. Nucleotide analysis of human lymphocytes indicate that inhibition of proliferation by BCX-1777 correlates with dGTP levels in the cells. BCX-1777 has excellent oral bioavailability (63%) in mice. At a single dose of 10 mg/kg in mice, BCX-1777 elevates dGuo to approximately 5 microM. BCX-1777 was not effective in mouse T cell models such as delayed type hypersensitivity (DTH) and splenomegaly because mouse T-cells do not accumulate dGTP as do human T-cells. However, in the human peripheral blood lymphocyte severe combined immunodeficiency (hu-PBL-SCID) mouse model, BCX-1777 was effective in prolonging the life span 2-fold or more. This is the first known example of a PNP inhibitor that elevates dGuo in mice similar to the levels observed in PNP-deficient patients. Furthermore, these dGuo levels are also required for in vitro T-cell inhibition by BCX-1777. Thus, BCX-1777 represents a novel class of selective immunosuppressive agents that could have therapeutic utility in various T-cell disorders. PMID- 11407315 TI - Primary immune system effects of the orally administered cyclopentane neuraminidase inhibitor RWJ-270201 in influenza virus-infected mice. AB - The cyclopentane derivative [1S,2S,3R,4R]-3-[(1S)-1-(acetylamino)-2- ethylbutyl] 4-[(aminoiminomethyl)amino]-2-hydroxy-cyclopentanecarboxylic acid (RWJ-270201) has been previously reported to be a potent and selective inhibitor of influenza virus neuraminidase, and to inhibit infections with this virus in vitro, in mice, and in clinical challenge studies. The effect of oral gavage therapy of 100 mg/kg/day of RWJ-270201 administered twice daily for 5 days beginning 16 h prior to virus exposure, on various immune factors of importance in response to primary influenza infection was determined in mice infected with influenza A/Shangdong/09/93 (H3N2) virus. Spleens taken from the mice 2 h after termination of treatment were processed for cytotoxic T lymphocytes (CTL) and natural killer (NK) cell activity and for enumeration of macrophages, T, T-helper, T suppressor/cytotoxic, and B cells. Saline-treated mice and normal mice were run in parallel. Treatment had no significant effect on any immune parameter. In a second experiment, mice infected with influenza A/NWS/33 (H1N1) were treated similarly with RWJ-270201 beginning 4 h pre-virus exposure. Treatment prevented any deaths from occurring, and markedly lessened arterial oxygen decline, lung consolidation, and lung virus titers. The mice developed mean neutralizing antibody (NA) titers of 1:592, and six of seven rechallenged mice resisted rechallenge with the same virus, indicating the initial virus-inhibitory effect also did not prevent the animals from developing an adequate humoral immune response to the virus. PMID- 11407316 TI - Tacrolimus suppressed the production of cytokines involved in atopic dermatitis by direct stimulation of human PBMC system. (Comparison with steroids). AB - Tacrolimus (FK506) ointment showed remarkable efficacy against atopic dermatitis in animal models and clinical trials. The suppressive effect of tacrolimus on the production of the cytokines involved in atopic dermatitis (IL-2, IL-3, IL-4, IL 5, IFN-gamma and GM-CSF) from human peripheral blood mononuclear cells (PBMC) was investigated. We constructed a new cytokine production system in which T cells are activated by direct stimulation in vitro with anti-CD3/CD2 or anti-CD3/CD28 antibody combination. Tacrolimus inhibited the production of these cytokines by both stimulations. In a comparative study with steroids (alclometasone dipropionate and betamethason valerate) in anti-CD3/CD2 system, tacrolimus and both steroids inhibited Th1 cytokines (IL-2, IFN-gamma), Th2 cytokines (IL-4, IL 5) and IL-3, GM-CSF (produced by both Th1 and Th2). The suppressive effect of tacrolimus on cytokine production was stronger than that of alclometasone dipropionate and equal to or stronger than that of betamethason valerate. The effective dose of tacrolimus (IC50, 0.02-0.11 ng/ml) is almost the same as for Th1 and Th2 cytokines, and 1 ng/ml of tacrolimus suppressed all cytokines completely. These results suggest that tacrolimus suppresses the allergic cytokines from T cells, and that tacrolimus ointment is effective against atopic dermatitis through the inhibition of cytokine production. PMID- 11407317 TI - Sex hormones as immunomodulators in health and disease. AB - In addition to their effects on sexual differentiation and reproduction, sex hormones influence the immune system. This results in a gender dimorphism in the immune function with females having higher immunoglobulin levels and mounting stronger immune responses following immunization or infection than males. The greater immune responsiveness in females is also evident in their increased susceptibility to autoimmune diseases. However, a clear understanding of the myriad of effects that sex hormones have on the immune system is lacking. Studies in normal mice show that estrogen treatment induces polyclonal B cell activation with increased expression of autoantibodies characteristic of autoimmune diseases. Several mechanisms appear to contribute to the break in tolerance and the increase in plasma cell activity including a reduction of the mass of the bone marrow and the thymus, the emergence of sites of extramedullary hematopoiesis and altered susceptibility of B cells to cell death. In addition, sex hormone levels in both humans and experimental models correlated with the activity of their cytokine-secreting cells indicating that sex hormones influence the cytokine milieu and suggesting that altered sex hormonal levels in autoimmune patients contribute to the skewed cytokine milieu characteristic of systemic lupus erythematosus (SLE). While sex hormones alone do not cause autoimmune disease, abnormal hormone levels may provide the stage for other factors (genetic, infectious) to trigger disease. Understanding the physiology of the interaction between sex hormones and immune function and its potential pathological consequences may provide insight into the autoimmune diseases and new directions for their treatment. PMID- 11407318 TI - Estrogen, prolactin, and autoimmunity: actions and interactions. AB - Estrogen and prolactin have a reciprocal endocrinologic relationship and both hormones have pleiotropic effects on the immune system. Despite the presence of a number of confounding variables, these hormones modulate autoimmunity; however, mechanisms by which this modulation occurs remain obscure. Estrogen appears to suppress cell-mediated and augment humoral-based immunity. Prolactin appears to stimulate both cell and humoral-based immunity. Both hormones have been shown to modulate IFN gamma secretion. Similar evidence in experimental models, human autoimmune disease, and during pregnancy in autoimmune disease patients suggests disparate effects of estrogen and prolactin on autoimmune responses and disease pathogenesis. In the NZB x NZW F1 mouse model of lupus, prolactin accelerates disease expression, whereas estrogen, devoid of its prolactin stimulating properties, is immunosuppressive and inhibits IL-2 production. Estrogen, because of its endocrinologic and immune effects, may directly or indirectly stimulate or inhibit immune responses. These dichotomous effects have limited its successful pharmacologic manipulation in human autoimmune disease with estrogen compounds, tamoxifen, oral contraceptives, antigonadotropic agents, or ovulation induction regimens. In contrast, reduction of immunostimulatory concentrations of prolactin with bromocriptine has successfully suppressed development or expression of murine and human autoimmune disease. Further investigation into actions and interactions of estrogen and prolactin with autoimmunity will provide a better understanding of the female preponderance of autoimmunity and facilitate a more rational approach to hormonal immunotherapy. PMID- 11407319 TI - Web alert. PMID- 11407320 TI - Web alert. PMID- 11407321 TI - Web alert. PMID- 11407322 TI - Web alert. PMID- 11407323 TI - Muller cells in retinal health and disease. PMID- 11407324 TI - Microbicides 2000 conference. Panel discussion: streamlining the process and regulatory issues. Microbicide research-streamlining the process, setting the stage. PMID- 11407325 TI - Microbicides 2000 conference. Panel discussion: streamlining the process and regulatory issues. Clinical trials and regulatory issues in developing countries. PMID- 11407326 TI - Microbicides 2000 conference. Panel discussion: streamlining the process and regulatory issues. PMID- 11407327 TI - Development of microbicides: WHO's perspective. PMID- 11407328 TI - The twin stories of Hanumanth Rao and Moses Ezekiel: the impact of reproductive health technologies on individual lives. PMID- 11407329 TI - [A survey of some new drugs, 2000]. PMID- 11407330 TI - [A retrospective in vivo study of Sonicsys approx restorations]. AB - The purpose of this retrospective study was to determine the longevity and quality of Class II resin composite restorations with Sonicsys approx inserts placed in private practice. In 79 patients 213 composite restorations with margins in enamel and in dentin were evaluated clinically using a modified version of established USPHS-Criteria. The mean time in clinical service was 18+ 8 months. The majority of the restorations investigated were rated as "good" (Alpha) or "clinically acceptable" (Bravo). In more than 90% sufficient proximal contact points were evaluated. Seven restorations had no contact to their adjacent tooth. Only one failed after an average period of 18 months in clinical service because of loss of the ceramic insert and was replaced. It is concluded that the composite restoration in combination with Sonicsys insert is an appropriate system for the restoration of Class II lesions in premolar and permanent molar teeth. PMID- 11407331 TI - [Modification of some occlusion concepts in implant dentistry: thoughts inspired by clinical experience]. AB - Implantology is nowadays a common tool in dentistry. While studying a case and selecting the future restoration, implants have to be considered when possible. Due to the functional differences between an implant and a natural tooth, the classical occlusion principles need to be considered. The prostheses have to be built according to the magnitude and direction of the forces that will be applied on the abutments. The prosthetic work and the occlusal equilibration are different in a patient with implants than in a patient with natural dentition. The occlusal concept will not be the same in the case of single implants surrounded by natural teeth, in the case of a mixed implant-tooth-supported fixed reconstruction and in the case of a full fixed bridge on implants. The purpose of this paper is to discuss the clinical consequences of these differences. PMID- 11407332 TI - Carbapenem-resistant Klebsiella pnuemoniae in Singapore producing IMP-1 beta lactamase and lacking an outer membrane protein. PMID- 11407333 TI - The power of modern computing and the internet in the delivery and evaluation of cardiac care. PMID- 11407334 TI - Virtual reality, telemedicine, distance care and education. PMID- 11407335 TI - Surgical management of thrombotic acute intestinal ischemia. AB - OBJECTIVE: To evaluate the University of Kentucky experience in treating acute intestinal ischemia to elucidate factors that contribute to survival. SUMMARY BACKGROUND DATA: Acute intestinal ischemia is reported to have a poor prognosis, with survival rates ranging from 0% to 40%. This is based on several reports, most of which were published more than a decade ago. Remarkably, there is a paucity of recent studies that report on current outcome for acute mesenteric ischemia. METHODS: A comparative retrospective analysis was performed on patients who were diagnosed with acute intestinal ischemia between May 1993 and July 2000. Patients were divided into two cohorts: nonthrombotic and thrombotic causes. The latter cohort was subdivided into three etiologic subsets: arterial embolism, arterial thrombosis, and venous thrombosis. Patient demographics, clinical characteristics, risk factors, surgical procedures, and survival were analyzed. Survival was compared with a collated historical series. RESULTS: Acute intestinal ischemia was diagnosed in 170 patients. The etiologies were nonthrombotic (102/170, 60%), thrombotic (58/170, 34%), or indeterminate (10/170, 6%). In the thrombotic cohort, arterial embolism accounted for 38% (22/58) of the cases, arterial thrombosis for 36% (21/58), and venous thrombosis for 26% (15/58). Patients with venous thrombosis were younger. Venous thrombosis was observed more often in men; arterial thrombosis was more frequent in women. The survival rate was 87% in the venous thrombosis group versus 41% and 38% for arterial embolism and thrombosis, respectively. Compared with the collated historical series, the survival rate was 52% versus 25%. CONCLUSIONS: These results indicate that the prognosis for patients with acute intestinal ischemia is substantially better than previously reported. PMID- 11407336 TI - Evolution of a multidisciplinary clinical pathway for the management of unstable patients with pelvic fractures. AB - OBJECTIVE: To determine whether the evolution of the authors' clinical pathway for the treatment of hemodynamically compromised patients with pelvic fractures was associated with improved patient outcome. SUMMARY BACKGROUND DATA: Hemodynamically compromised patients with pelvic fractures present a complex challenge. The multidisciplinary trauma team must control hemorrhage, restore hemodynamics, and rapidly identify and treat associated life-threatening injuries. The authors developed a clinical pathway consisting of five primary elements: immediate trauma attending surgeon's presence in the emergency department, early simultaneous transfusion of blood and coagulation factors, prompt diagnosis and management of associated life-threatening injuries, stabilization of the pelvic girdle, and timely insinuation of pelvic angiography and embolization. The addition of two orthopedic pelvic fracture specialists led to a revision of the pathway, emphasizing immediate emergency department presence of the orthopedic trauma attending to provide joint decision making with the trauma surgeon, closing the pelvic volume in the emergency department, and using alternatives to traditional external fixation devices. METHODS: Using trauma registry and blood bank records, the authors identified pelvic fracture patients receiving blood transfusions in the emergency department. They analyzed patients treated before versus after the May 1998 revision of the clinical pathway. RESULTS: A higher proportion of patients in the late period had blood pressure less than 90 mmHg (52% vs. 35%). In the late period, diagnostic peritoneal lavage was phased out in favor of torso ultrasound as a primary triage tool, and pelvic binding and C-clamp application largely replaced traditional external fixation devices. The overall death rate decreased from 31% in the early period to 15% in the later period, as did the rate of deaths from exsanguination (9% to 1%), multiple organ failure (12% to 1%), and death within 24 hours (16% to 5%). CONCLUSIONS: The evolution of a multidisciplinary clinical pathway, coordinating the resources of a level 1 trauma center and directed by joint decision making between trauma surgeons and orthopedic traumatologists, has resulted in improved patient survival. The primary benefits appear to be in reducing early deaths from exsanguination and late deaths from multiple organ failure. PMID- 11407337 TI - Difficult flotation of a pulmonary artery catheter: echocardiographic diagnosis. PMID- 11407338 TI - [Evolution of the prevalence of obesity in France]. PMID- 11407339 TI - [Control of 'peaks and valleys']. PMID- 11407340 TI - Proceedings of the 4th International Pediatric Radiology Meeting. Paris, France, May 28-June 1, 2001. PMID- 11407341 TI - The outbreak of West Nile virus infection in the New York City area in 1999. AB - BACKGROUND: In late August 1999, an unusual cluster of cases of meningoencephalitis associated with muscle weakness was reported to the New York City Department of Health. The initial epidemiologic and environmental investigations suggested an arboviral cause. METHODS: Active surveillance was implemented to identify patients hospitalized with viral encephalitis and meningitis. Cerebrospinal fluid, serum, and tissue specimens from patients with suspected cases underwent serologic and viral testing for evidence of arboviral infection. RESULTS: Outbreak surveillance identified 59 patients who were hospitalized with West Nile virus infection in the New York City area during August and September of 1999. The median age of these patients was 71 years (range, 5 to 95). The overall attack rate of clinical West Nile virus infection was at least 6.5 cases per million population, and it increased sharply with age. Most of the patients (63 percent) had clinical signs of encephalitis; seven patients died (12 percent). Muscle weakness was documented in 27 percent of the patients and flaccid paralysis in 10 percent; in all of the latter, nerve conduction studies indicated an axonal polyneuropathy in 14 percent. An age of 75 years or older was an independent risk factor for death (relative risk adjusted for the presence or absence of diabetes mellitus, 8.5; 95 percent confidence interval, 1.2 to 59.1), as was the presence of diabetes mellitus (age-adjusted relative risk, 5.1; 95 percent confidence interval, 1.5 to 17.3). CONCLUSIONS: This outbreak of West Nile meningoencephalitis in the New York City metropolitan area represents the first time this virus has been detected in the Western Hemisphere. Given the subsequent rapid spread of the virus, physicians along the eastern seaboard of the United States should consider West Nile virus infection in the differential diagnosis of encephalitis and viral meningitis during the summer months, especially in older patients and in those with muscle weakness. PMID- 11407342 TI - Hematopoietic engraftment and survival in adult recipients of umbilical-cord blood from unrelated donors. AB - BACKGROUND: Umbilical-cord blood from unrelated donors who are not HLA-identical with the recipients can restore hematopoiesis after myeloablative therapy in children. We studied the use of transplantation of umbilical-cord blood to restore hematopoiesis in adults. METHODS: Sixty-eight adults with life threatening hematologic disorders received intensive chemotherapy or total-body irradiation and then transplants of HLA-mismatched umbilical-cord blood. We evaluated the outcomes in terms of hematologic reconstitution, the occurrence of acute and chronic graft-versus-host disease (GVHD), relapses, and event-free survival. RESULTS: Of the 68 patients, 48 (71 percent) received grafts of umbilical-cord blood that were mismatched for two or more HLA antigens. Of the 60 patients who survived 28 days or more after transplantation, 55 had neutrophil engraftment at a median of 27 days (range, 13 to 59). The estimated probability of neutrophil recovery in the 68 patients was 0.90 (95 percent confidence interval, 0.85 to 1.0). The presence of a relatively high number of nucleated cells in the umbilical-cord blood before it was frozen was associated with faster recovery of neutrophils. Severe acute GVHD (of grade III or IV) occurred in 11 of 55 patients who could be evaluated within the first 100 days after transplantation. Chronic GVHD developed in 12 of 33 patients who survived for more than 100 days after transplantation. The median follow-up for survivors was 22 months (range, 11 to 51). Of the 68 patients, 19 were alive and 18 of these (26 percent) were disease-free 40 months after transplantation. The presence of a high number of CD34+ cells in the graft was associated with improved event-free survival (P=0.05). CONCLUSIONS: Umbilical-cord blood from unrelated donors can restore hematopoiesis in adults who receive myeloablative therapy and is associated with acceptable rates of severe acute and chronic GVHD. PMID- 11407343 TI - Identification of a gene responsible for familial Wolff-Parkinson-White syndrome. AB - BACKGROUND: The Wolff-Parkinson-White syndrome, with a prevalence in Western countries of 1.5 to 3.1 per 1000 persons, causes considerable morbidity and may cause sudden death. We identified two families in which the Wolff-Parkinson-White syndrome segregated as an autosomal dominant disorder. METHODS: We studied 70 members of the two families (57 in Family 1 and 13 in Family 2). The subjects underwent 12-lead electrocardiography and two-dimensional echocardiography. Genotyping mapped the gene responsible to 7q34-q36, a locus previously identified to be responsible for an inherited form of Wolff-Parkinson-White syndrome. Candidate genes were identified, sequenced, and analyzed in normal and affected family members to identify the disease-causing gene. RESULTS: A total of 31 members (23 from Family 1 and 8 from Family 2) had the Wolff-Parkinson-White syndrome. Affected members of both families had ventricular preexcitation with conduction abnormalities and cardiac hypertrophy. The maximal combined two-point lod score was 9.82 at a distance of 5 cM from marker D7S636, which confirmed the linkage of the gene in both families to 7q34-q36. Haplotype analysis indicated that there were no alleles in common in the two families at this locus, suggesting that the two families do not have a common founder. We identified a missense mutation in the gene that encodes the gamma2 regulatory subunit of AMP activated protein kinase (PRKAG2). The mutation results in the substitution of glutamine for arginine at residue 302 in the protein. CONCLUSIONS: The identification of this genetic defect has important implications for elucidating the pathogenesis of ventricular preexcitation. Further understanding of how this molecular defect leads to supraventricular arrhythmias could influence the development of specific therapies for other forms of supraventricular arrhythmia. PMID- 11407344 TI - Neonatal pulmonary hypertension--urea-cycle intermediates, nitric oxide production, and carbamoyl-phosphate synthetase function. AB - BACKGROUND: Endogenous production of nitric oxide is vital for the decrease in pulmonary vascular resistance that normally occurs after birth. The precursor of nitric oxide is arginine, a urea-cycle intermediate. We hypothesized that low concentrations of arginine would correlate with the presence of persistent pulmonary hypertension in newborns and that the supply of this precursor would be affected by a functional polymorphism (the substitution of asparagine for threonine at position 1405 [T1405N]) in carbamoyl-phosphate synthetase, which controls the rate-limiting step of the urea cycle. METHODS: Plasma concentrations of amino acids and genotypes of the carbamoyl-phosphate synthetase variants were determined in 65 near-term neonates with respiratory distress. Plasma nitric oxide metabolites were measured in a subgroup of 10 patients. The results in infants with pulmonary hypertension, as assessed by echocardiography, were compared with those in infants without pulmonary hypertension. The frequencies of the carbamoyl-phosphate synthetase genotypes in the study population were assessed for Hardy-Weinberg equilibrium. RESULTS: As compared with infants without pulmonary hypertension, infants with pulmonary hypertension had lower mean (+/-SD) plasma concentrations of arginine (20.2+/-8.8 vs. 39.8+/-17.0 micromol per liter, P<0.001) and nitric oxide metabolites (18.8+/-12.7 vs. 47.2+/ 11.2 micromol per liter, P=0.05). As compared with the general population, the infants in the study had a significantly skewed distribution of the genotypes for the carbamoyl-phosphate synthetase variants at position 1405 (P<0.005). None of the infants with pulmonary hypertension were homozygous for the T1405N polymorphism. CONCLUSIONS: Infants with persistent pulmonary hypertension have low plasma concentrations of arginine and nitric oxide metabolites. The simultaneous presence of diminished concentrations of precursors and breakdown products suggests that inadequate production of nitric oxide is involved in the pathogenesis of neonatal pulmonary hypertension. Our preliminary observations suggest that the genetically predetermined capacity of the urea cycle--in particular, the efficiency of carbamoyl-phosphate synthetase--may contribute to the availability of precursors for nitric oxide synthesis. PMID- 11407345 TI - Images in clinical medicine. Neonatal pneumomediastinum. PMID- 11407346 TI - Clinical practice. Noninvasive tests in patients with stable coronary artery disease. PMID- 11407347 TI - The irritable bowel syndrome. PMID- 11407348 TI - Case records of the Massachusetts General Hospital. Weekly clinicopathological exercises. Case 18-2001. A 19-month-old boy with fever and soft-tissue masses. PMID- 11407349 TI - West Nile virus encephalitis in America. PMID- 11407350 TI - Hematopoietic stem-cell transplants using umbilical-cord blood. PMID- 11407351 TI - A molecular basis for Wolff-Parkinson-White syndrome. PMID- 11407352 TI - Intussusception and an oral rotavirus vaccine. PMID- 11407353 TI - Intussusception and an oral rotavirus vaccine. PMID- 11407354 TI - Green tea and gastric cancer in Japan. PMID- 11407355 TI - Captopril-augmented renal scanning. PMID- 11407356 TI - Aspiration pneumonia. PMID- 11407357 TI - Aspiration pneumonia. PMID- 11407358 TI - Aspiration pneumonia. PMID- 11407359 TI - Case 3-2001: unilateral visual loss. PMID- 11407360 TI - Case 3-2001: unilateral visual loss. PMID- 11407361 TI - Creation of a double chimera after the transplantation of umbilical-cord blood from two partially matched unrelated donors. PMID- 11407362 TI - Response to "telomere shortening with aging in human liver". PMID- 11407363 TI - Response to "increased bioactivity of rat atrial extracts: relation to aging and blood pressure regulation". PMID- 11407364 TI - Why are doctors so unhappy? Calibre of people recruited to medicine may be too high for the job. PMID- 11407365 TI - Why are doctors so unhappy? Lack of leadership may be a contributory cause. PMID- 11407366 TI - Why are doctors so unhappy? Reducing working hours might help. PMID- 11407367 TI - Why are doctors so unhappy? Are we unhappy? PMID- 11407368 TI - Why are doctors so unhappy? Workload has increased dramatically. PMID- 11407369 TI - Why are doctors so unhappy? General practitioners need to negotiate new contract. PMID- 11407370 TI - Why are doctors so unhappy? Consultant based service is needed. PMID- 11407371 TI - Why are doctors so unhappy? Medical profession must unite to address problems. PMID- 11407372 TI - Why are doctors so unhappy? Unhappiness will be defeated when doctors accept full social responsibility. PMID- 11407373 TI - Why are doctors so unhappy? Analysis is timely but blinkered. PMID- 11407374 TI - Why are doctors so unhappy? Exit from eternal triangle of perpetrator-victim caretaker is needed. PMID- 11407375 TI - Why are doctors so unhappy? Doctors have only themselves to blame. PMID- 11407376 TI - Why are doctors so unhappy? Doctor support groups can be a lifeline. PMID- 11407377 TI - Glucose tolerance in adults after prenatal exposure to famine. PMID- 11407378 TI - Glucose tolerance in adults after prenatal exposure to famine. PMID- 11407379 TI - Glucose tolerance in adults after prenatal exposure to famine. PMID- 11407380 TI - Glucose tolerance in adults after prenatal exposure to famine. PMID- 11407381 TI - Helminthiasis and HIV vaccine efficacy. PMID- 11407382 TI - Transmission of cytomegalovirus. PMID- 11407383 TI - Transmission of cytomegalovirus. PMID- 11407384 TI - Differences in invasive pneumococcal serotypes. PMID- 11407385 TI - Recurrent syncope and presyncope. PMID- 11407386 TI - Recurrent syncope and presyncope. PMID- 11407387 TI - Variant esp gene in vancomycin-sensitive Enterococcus faecium. PMID- 11407388 TI - Mitochondrial toxic effects and ribavirin. PMID- 11407389 TI - Mitochondrial toxic effects and ribavirin. PMID- 11407390 TI - Genetic susceptibility and familial malignant mesothelioma. PMID- 11407391 TI - Tetanus in the elderly: a forgotten illness. PMID- 11407392 TI - Animal tests after the genome. PMID- 11407393 TI - Animal tests after the genome. PMID- 11407394 TI - Animal tests after the genome. PMID- 11407395 TI - Street crime as a treatment for migraine. PMID- 11407396 TI - Genomic and serological diversity of bovine viral diarrhea virus in Japan. AB - Genomic properties of 62 field isolates of bovine viral diarrhea virus (BVDV) collected from 1974 to 1999 in Japan were investigated. The 5' untranslated region (UTR) was amplified by reverse transcriptase-polymerase chain reaction (RT PCR) and the 244 to 247 base nucleotide sequences were determined. Serological properties were also characterized by the cross-neutralization test using antisera against the representative strain of the classified genotype. Using phylogenetic tree analysis, BVDV 1 was subdivided into two major clusters, BVDV 1a (29 isolates) and BVDV-1b (27 isolates). In group 1a, 3 differed from the other viruses, and were classified in a branch assigned as 1a'. However, 4 isolates (So CP/75, 190 CP, 190 NCP and KS86-1-NCP) could not be assigned to group 1a or 1b. In comparison with the published sequence data, KS86-1-NCP, 190 CP and 190 NCP were similar to the Southern Africa isolates that have recently been proposed as BVDV 1c. The 5' UTR sequence of So CP/75 was unique among those of BVDV 1, suggesting that the isolate should be classified into a new genotype. Only 2 out of 62 isolates collected in 1989 and 1990 were identified as BVDV 2. The results of the cross-neutralization test strongly supported these data, suggesting a close correlation between the 5' UTR sequence and the antigenicity of BVDV. PMID- 11407397 TI - Dramatic response of teratoma-associated non--germ-cell cancer with all-trans retinoic acid in a patient with nonseminomatous germ cell tumor. AB - A patient with nonseminomatous germ cell cancer, treated with standard chemotherapy, subsequently developed a pathologically confirmed metastatic undifferentiated adenocarcinoma (non-germ-cell elements) arising from residual teratoma. Disease was present in both lobes of the liver and was deemed unresectable at the time of presentation. The patient was treated on a National Cancer Institute-sponsored institutional protocol with all-trans retinoic acid. After 60 days of oral therapy at a dose of 150 mg/m2/d (50 mg/m2 three times daily), the patient was found to have complete radiologic resolution of his hepatic metastases. He subsequently underwent surgery and his complete response was pathologically confirmed. PMID- 11407398 TI - Images in clinical medicine. Zidovudine myopathy. PMID- 11407399 TI - Histopathology and cytopathology of white spot syndrome virus (WSSV) in cultured Penaeus monodon from peninsular Malaysia with emphasis on pathogenesis and the mechanism of white spot formation. AB - Since 1994, white spot syndrome virus (WSSV) has been detected in cultured shrimp Penaeus monodon in Peninsular Malaysia. The gross signs, target organs and histo cytopathology for the viral infection were studied and it was found to infect most organs and tissues including oocytes, but not hepatopancreatocytes and epithelial cells of the midgut, which were regarded as refractory tissues. Based on a time-sequence of ultrastructural cytopathology, 4 cytopathic profiles and 6 phases of viral morphogenesis were described. The virions were elliptical to short rods with trilamilar envelopes that measured 305 +/- 30 x 127 +/- 11 nm. Viral nucleosomes were often present singly in infected nuclei and were associated with the early stages of viral replication. The structure of WSSV pathognomonic white, cuticular lesions was examined at the microscopic and ultrastructural levels and the mechanism of their formation appeared to be related to the disruption of exudate transfer from epithelial cells to the cuticle via cuticular pore canals. PMID- 11407400 TI - Studies on effective PCR screening strategies for white spot syndrome virus (WSSV) detection in Penaeus monodon brooders. AB - We re-tested stored (frozen) DNA samples in 5 independent polymerase chain reaction (PCR) replicates and confirmed that equivocal test results from a previous study on white spot syndrome virus (WSSV) in brooders and their offspring arose because amounts of WSSV DNA in the test samples were near the sensitivity limits of the detection method. Since spawning stress may trigger WSSV replication, we also captured a fresh batch of 45 brooders for WSSV PCR testing before and after spawning. Replicates of their spawned egg batches were also WSSV PCR tested. For these 45 brooders, WSSV prevalence before spawning was 67% (15/45 1-step PCR positive, 15/45 2-step PCR positive and 15/45 2-step PCR negative). Only 27 (60%) spawned successfully. Of the successful spawners, 56% were WSSV PCR positive before spawning and 74% after. Brooders (15) that were heavily infected (i.e. 1-step PCR positive) when captured mostly died within 1 to 4 d, but 3 (20%) did manage to spawn. All their egg batch sub-samples were 1-step PCR positive and many failed to hatch. The remaining 30 shrimp were divided into a lightly infected group (21) and a 2-step PCR negative group (9) based on replicate PCR tests. The spawning rates for these 2 groups were high (81 and 78%, respectively). None of the negative spawners (7) became WSSV positive after spawning and none gave egg samples positive for WSSV. In the lightly infected group (21), 6 brooders were 2-step WSSV PCR negative and 15 were 2-step WSSV PCR positive upon capture. However, all of them were WSSV PCR positive in replicate tests and after spawning or death. Four died without spawning. The remaining 17 spawned but only 2 gave egg samples PCR negative for WSSV. The other 15 gave PCR positive egg samples, but they could be divided into 2 spawner groups: those (7) that became heavily infected (i.e. 1-step PCR positive) after spawning and those (8) that remained lightly infected (i.e. became or remained 2-step PCR positive only). Of the brooders that became heavily infected after spawning, almost all egg sample replicates (91 %) tested 2-step PCR positive. One brooder even gave heavily infected (i.e. 1-step PCR positive) egg samples. For the brooders that remained lightly infected after spawning, only 27% of the egg sample replicates were 2-step PCR positive. Based on these results, we recommend that to avoid false negatives in WSSV PCR brooder tests screening tests should be delayed until after spawning. We also recommend, with our PCR detection system, discarding all egg batches from brooders that are 1-step PCR positive after spawning. On the other hand, it may be possible with appropriate monitoring to use eggs from 2 step PCR positive brooders for production of WSSV-free or lightly infected postlarvae. These may be used to stock shrimp ponds under low-stress rearing conditions. PMID- 11407401 TI - Results from black tiger shrimp penaeus monodon culture ponds stocked with postlarvae PCR-positive or -negative for white-spot syndrome virus (WSSV). AB - Commercial, intensive, earthen shrimp ponds (188) in southern Thailand were stocked with postlarvae (PL) of Penaeus monodon that had tested positive or negative for white-spot syndrome virus (WSSV) infection by polymerase chain reaction (PCR) assay. All the PL were grossly healthy. At 2 wk intervals after stocking, shrimp from each pond were examined for gross WSSV lesions and tested for WSSV by PCR. Shrimp from all the ponds stocked with WSSV-PCR-positive PL (Group 0, n = 43) eventually showed gross signs of white-spot disease (WSD) at an average of 40 d after stocking. Of the remaining ponds stocked with WSSV-PCR negative PL (n = 145), some remained WSSV-PCR-negative throughout the study (Group 5, n = 52), while others (93) became WSSV-PCR-positive after stocking, during the first month (Group 1, n = 23), second month (Group 2, n = 40), third month (Group 3, n = 24), or fourth month (Group 4, n = 6). Crop failure was defined as a pond drain or forced harvest before 14 wk or 98 d of cultivation. For Group 0 the proportion of ponds failing was 0.953, while it was only 0.019 for Group 5. Thus, the relative risk of failure for Group 0 was approximately 50 times that of Group 5. The relative risk of failure for Group 0 was also 3 times that for ponds stocked with WSSV-PCR-negative PL. Obviously, not all WSSV outbreaks resulted in crop failure. Of the 93 ponds stocked with PCR-negative PL that later yielded WSSV-PCR-positive shrimp, 53% reached successful harvest. The study showed that PCR screening of PL and rejection of WSSV-positive batches before stocking could greatly improve the chances of a successful harvest. PMID- 11407402 TI - Evaluation of the protective immunogenicity of the N, P, M, NV and G proteins of infectious hematopoietic necrosis virus in rainbow trout oncorhynchus mykiss using DNA vaccines. AB - The protective immunogenicity of the nucleoprotein (N), phosphoprotein (P), matrix protein (M), non-virion protein (NV) and glycoprotein (G) of the rhabdovirus infectious hematopoietic necrosis virus (IHNV) was assessed in rainbow trout using DNA vaccine technology. DNA vaccines were produced by amplifying and cloning the viral genes in the plasmid pCDNA 3.1. The protective immunity elicited by each vaccine was evaluated through survival of immunized fry after challenge with live virus. Neutralizing antibody titers were also determined in vaccinated rainbow trout Oncorhynchus mykiss fry (mean weight 2 g) and 150 g sockeye salmon Oncorhynchus nerka. The serum from the 150 g fish was also used in passive immunization studies with naive fry. Our results showed that neither the internal structural proteins (N, P and M) nor the NV protein of IHNV induced protective immunity in fry or neutralizing antibodies in fry and 150 g fish when expressed by a DNA vaccine construct. The G protein, however, did confer significant protection in fry up to 80 d post-immunization and induced protective neutralizing antibodies. We are currently investigating the role of different arms of the fish immune system that contribute to the high level of protection against IHNV seen in vaccinated fish. PMID- 11407403 TI - High permissivity of the fish cell line SSN-1 for piscine nodaviruses. AB - Seventeen isolates of piscine nodavirus from larvae or juveniles of 13 marine fish species affected with viral nervous necrosis (VNN) were examined for their infectivity to a fish cell line SSN-1. Based on cytopathic effects (CPE) and virus antigen detection by fluorescent antibody technique (FAT) after incubation at 25 degrees C, the infectivity of these virus isolates was divided into 4 groups. Group 1, including 9 virus isolates from 4 species of grouper, 2 species of sea bass, barramundi, rock porgy, and Japanese flounder showed CPE characterized by rounded, granular cells with heavy cytoplasmic vacuoles within 3 d post-incubation (p.i.), and the monolayer partially or completely disintegrated over 3 to 6 d p.i. Scattered FAT-positive cells appeared at 3 h p.i. and spread through the cell sheet with an increasing fluorescence signal over 24 h p.i. Group 2, consisting of 3 virus isolates from striped jack, induced CPE with thin or rounded, granular, refractile cells without conspicuous vacuole formation, and extensive FAT-positive reaction was observed in a time course similar to that of Group 1. Cells inoculated with Group 3 (1 isolate from tiger puffer) developed no distinct CPE but viral infection was evidenced by localized FAT-positive cells. There were no FAT-positive cells in Group 4, which included 4 isolates from Japanese flounder, Pacific cod and Atlantic halibut. However, when incubation was performed at 20 degrees C, the SSN-1 cells inoculated with the Group 3 isolate showed CPE similar to that of Group 1 and extensive FAT-positive reaction. Evidence of virus proliferation at 20 degrees C was also obtained in Group 4 isolates. The virus titers in the infected fish varied from 10(11) to 10(16) tissue culture infectious dose (TCID50) g(-1) of fish. There is a good correlation between these infectivities to the SSN-1 cells and the coat protein gene genotypes of the isolates. The present results indicate that SSN-1 cells are useful for propagating and differentiating genotypic variants of piscine nodavirus. PMID- 11407404 TI - Skin tumours in Pleuronectes obscurus (Pleuronectidae) represent a complex combination of epidermal papilloma and rhabdomyosarcoma. AB - In the present work we describe the histology of skin tumours of the black plaice Pleuronectes obscurus from Amursky Bay, the Sea of Japan. The epidermis forms numerous papillary folds protruding above the skin surface and supported by delicate branches of connective tissue. This type of neoplasm is classified as epidermal papilloma. The occurrence of severe epidermal hyperplasia and disturbance of the histoarchitecture in some areas, invasion of the adjacent connective tissues by epithelial cells, dystrophic changes of the epithelial cells, and the occurrence of a large number of mitoses point to an increasing malignancy of the papilloma. Moreover, areas with skeletal-muscle differentiation were found within skin tumours. Among the myogenic cells, features of normal somatic myogenesis were observed along with signs of abnormality of this process, suggesting a disturbance of myogenic differentiation. Cellular polymorphism among myogenic cells and invasion of the skin by neoplastic cells are evidence of the malignant character of this type of tumour and allow us to classify it as rhabdomyosarcoma. Due to the position of tumours in the skin, they are ectopic rhabdomyosarcomas. In the skin tumours, atypical small and large rounded cells were identified, the latter having previously been described in flatfish as X cells. The origin of these cells is discussed and the assumption is put forward that small and large rounded cells can be regarded as cellular elements of rhabdomyosarcomas. PMID- 11407405 TI - Lectin-reactive components of the microsporidian Glugea plecoglossi and their relation to spore phagocytosis by head kidney macrophages of ayu Plecoglossus altivelis. AB - We investigated the reactivity of lectins to spores of Glugea plecoglossi from ayu Plecoglossus altivelis. Smear preparations of purified spores were treated with 8 kinds of lectins. Lectin blots were used to detect glycoproteins of spore lysates. In addition, lectin-treated spores were applied to head kidney macrophages of ayu, and the percentage of phagocytosis (PP) was calculated and compared with the control. Two lectins (ConA, WGA) reacted with the surface of the spores, and a major band (55 kDa) and some minor bands were visualized on blots after treatment with these. PP was decreased after ConA treatment. From these results, we suggest that G. plecoglossi spores can be phagocytized by ayu head kidney macrophages via ConA-reactive glycoprotein-mediated recognition. PMID- 11407406 TI - Massive infestation by Amyloodinium ocellatum (Dinoflagellida) of fish in a highly saline lake, Salton Sea, California, USA. AB - Persistent fish infestation by the parasitic dinoflagellate Amyloodinium ocellatum was found at a highly saline lake, Salton Sea, California, USA. The seasonal dynamics of the infestation of young tilapia was traced in 1997-1998. First appearing in May, it became maximal in June-August, decreased in October and was not detectable in November. Outbreak of the infestation and subsequent mortality of young fish was registered at the Sea at a water temperature and salinity of 40 degrees C and 46 ppt, respectively. Some aspects of the ultrastructure of parasitic trophonts of A. ocellatum and their location on the fish from different size groups are considered. The interactions of parasitological and environmental factors and their combined effect upon fish from the Salton Sea are discussed. PMID- 11407407 TI - Development of Anguillicola crassus (Dracunculoidea, Anguillicolidae) in experimentally infected Balearic congers Ariosoma balearicum (Anguilloidea, Congridae). AB - The development of Anguillicola crassus in experimentally infected Ariosoma balearicum (Anguilloidea, Congridae) kept in seawater was studied in the laboratory. In parallel trials the effect of water salinity on the development of larval A. crassus in European eels Anguilla anguilla was also investigated using eels kept in seawater of a salinity of 34 per thousand. Both eel species were orally inoculated with L3 larvae of A. crassus and then maintained for up to 3 mo at 18 degrees C in seawater. 110 d post infection, no adult but larval (L3 and L4) stages of A. crassus were detected in the swimbladder wall of Balearic congers, although this period of time was sufficient for the parasites to develop to the adult stage in European eel kept in seawater. The results presented suggest that the definitive host specificity of A. crassus comprises species of the family Anguillidae (i.e. the genus Anguilla), but not members of the Congridae. Theoretically however, A. balearicum might serve as a metaparatenic host. Factors determining the definitive host range of A. crassus remain to be elucidated. Water salinity does not seem to act as a factor affecting definitive host specificity once the parasite has become ingested by the eel. PMID- 11407408 TI - On feeling good and getting your way: mood effects on negotiator cognition and bargaining strategies. AB - Are happy people more likely to be cooperative and successful negotiators? On the basis of the Affect Infusion Model (AIM; Forgas, 1995a). Experiment 1 predicted and found that both good and bad moods had a significant mood-congruent effect on people's thoughts and plans, and on their negotiation strategies and outcomes in both interpersonal and intergroup bargaining. Experiment 2 replicated these results and also showed that mood effects were reduced for persons more likely to adopt motivated processing strategies (scoring high on machiavellianism and need for approval). Experiment 3 confirmed these effects and demonstrated that the mood of the opposition also produced more mood-congruent bargaining strategies and outcomes. The results are discussed in terms of affect priming influences on interpersonal behaviors, and the implications of these findings for real-life cognitive tasks and bargaining encounters are considered. PMID- 11407409 TI - Centrosome inheritance in insects: fertilization and parthenogenesis. AB - Centrosome biogenesis is unclear, although much structural and biochemical research has been performed in several experimental systems. An alternative model to study the assembly of functional centrosomes could be the process of zygotic centrosome formation at the beginning of embryonic development. Although it seems obvious that the sperm cell provides the centrosome at fertilization, some pieces of evidence are not in line with this point of view and give controversial results. Such an analysis could provide useful information if applied to a large variety of organisms. Since insects are a highly diverse group of organisms they provide a variety of models in which to study the process of centrosome reconstitution during fertilization. Moreover, many insect species reproduce by parthenogenesis, a special mode of reproduction in which embryonic development occurs without male contribution. Studies of unfertilized parthenogenetic eggs may therefore teach us much about the process of centrosome assembly in the absence of preexisting centrioles. PMID- 11407410 TI - Single molecule fluorescence burst detection of DNA fragments separated by capillary electrophoresis. AB - A method has been developed for detecting DNA separated by capillary gel electrophoresis (CGE) using single molecule photon burst counting. A confocal fluorescence microscope was used to observe the fluorescence bursts from single molecules of DNA multiply labeled with the thiazole orange derivative T06 as they passed through the approximately 2 micrometer diameter focused laser beam. Amplified photoelectron pulses from the photomultiplier are grouped into bins of 360-450 micros in duration, and the resulting histogram is stored in a computer for analysis. Solutions of M13 DNA were first flowed through the capillary at various concentrations, and the resulting data were used to optimize the parameters for digital filtering using a low-pass Fourier filter, selecting a discriminator level for peak detection, and applying a peak-calling algorithm. Statistical analyses showed that (i) the number of M13 molecules counted versus concentration was linear with slope = 1, (ii) the average burst duration was consistent with the expected transit time of a single molecule through the laser beam, and (iii) the number of detected molecules was consistent with single molecule detection. The optimized single molecule counting method was then applied to an electrophoretic separation of M13 DNA and to a separation of pBR 322 DNA from pRL 277 DNA. Clusters of discreet fluorescence bursts were observed at the expected appearance time of each DNA band. The autocorrelation function of these data indicated transit times that were consistent with the observed electrophoretic velocity. These separations were easily detected when only 50-100 molecules of DNA per band traveled through the detection region. This new detection technology should lead to the routine analysis of DNA in capillary columns with an on-column sensitivity of approximately 100 DNA molecules/band or better. PMID- 11407411 TI - Protein binding to the abscisic acid-responsive element is independent of VIVIPAROUS1 in vivo. AB - The plant hormone abscisic acid and the transcriptional activator VIVIPAROUS1 have a synergistic effect on transcription during embryo development. An abscisic acid-responsive element (ABRE) mediates induction by abscisic acid and VIVIPAROUS1, but the mechanism involved has not been determined. In this study, we explore the interaction between abscisic acid and VIVIPAROUS1 and its effect on the ABRE from the maize rab28 gene. In transient transformation experiments, abscisic acid stimulated transcription via several elements, whereas activation by VIVIPAROUS1 was mediated exclusively through the ABRE. In vivo footprinting showed only minor differences in binding to the ABRE between wild-type and VIVIPAROUS1-deficient embryos, suggesting that VIVIPAROUS1 stimulates transcription through the ABRE without major changes in protein-DNA interactions. A factor that bound to the ABRE in electrophoretic mobility shift assays was present at the same developmental stages as rab28 mRNA and had binding characteristics similar to those observed by in vivo footprinting. This suggests that the factor binds to the ABRE in the rab28 promoter in vivo. We discuss the constraints that our results put on the possible mechanism for action of VIVIPAROUS1 in vivo. PMID- 11407412 TI - Effects of a knowledge-based electronic patient record in adherence to practice guidelines. PMID- 11407413 TI - Oscillation and noise determine signal transduction in shark multimodal sensory cells. AB - Oscillating membrane potentials that generate rhythmic impulse patterns are considered to be of particular significance for neuronal information processing. In contrast, noise is usually seen as a disturbance which limits the accuracy of information transfer. We show here, however, that noise in combination with intrinsic oscillations can provide neurons with particular encoding properties, a discovery we made when recording from single electro-sensory afferents of a fish. The temporal sequence of the impulse trains indicates oscillations that operate near the spike-triggering threshold. The oscillation frequency determines the basic rhythm of impulse generation, but whether or not an impulse is actually triggered essentially depends on superimposed noise. The probability of impulse generation can be altered considerably by minor modifications of oscillation baseline and amplitude, which may underlie the exquisite sensitivity of these receptors to thermal and electrical stimuli. Additionally, thermal, but not electrical, stimuli alter the oscillation frequency, allowing dual sensory messages to be conveyed in a single spike train. These findings demonstrate novel properties of sensory transduction which may be relevant for neuronal signalling in general. PMID- 11407414 TI - Correlations between amino acid hydrophobicity scales and stain exclusion capacity of type 1 collagen fibrils. AB - The relationship between the negative staining band pattern of type 1 native collagen fibrils and the amino acid distribution along the fibril axis was studied by comparing averaged microdensitograms with theoretical traces calculated on the basis of different amino acid parameters. As well as the spatial parameter "bulkiness" (volume/length, ratio), various literature-reported scales of "hydrophobicity" were tested. Two "hydrophobicity" sets allowed a better fit with the actual patterns than "bulkiness" values. However, a general improvement in simulations was achieved by associating most "hydrophobicity" sets with the "bulkiness" set. These results suggest that amino acid "hydrophobicity" plays a key role in the appearance of negative staining patterns but a composite mechanism would seem to occur: the accessibility of available intermolecular interstices may be conditioned by molecular hindrance, corresponding to amino acid "bulkiness" as well as by water-repulsion effect, which correlates with amino acid "hydrophobicity." Moreover, a detailed comparison of actual and simulated patterns suggests that a modulation exists in the effectiveness of these two factors along each D-period according to the different molecular packing and concentration of hydrophobic amino acid clusters within overlap regions and gap regions, respectively. PMID- 11407415 TI - Analogical priming via semantic relations. AB - Research on semantic memory has often tacitly treated semantic relations as simple conduits for spreading activation between associated object concepts, rather than as integral components of semantic organization. Yet conceptual relations, and the role bindings they impose on the objects they relate, are central to such cognitive tasks as discourse comprehension, inference, problem solving, and analogical reasoning. The present study addresses the question of whether semantic relations and their bindings can influence access to semantic memory. The experiments investigated whether, and under what conditions, presenting a prime pair of words linked by 1 of 10 common semantic relations would facilitate processing of a target pair of words linked by the same relation. No effect was observed when participants merely read the prime; however, relational priming was observed under instructions to note and use the semantic relations. Participants were faster at making a lexical decision or naming a word on a related pair of target words when that pair was primed with an analogously related pair of words than when the prime pair consisted of either two unrelated words or two words linked by some other relation. This evidence of analogical priming suggests that under an appropriate strategic set, lexical decisions and naming latencies can be influenced by a process akin to analogical mapping. PMID- 11407416 TI - Social influences on reality-monitoring decisions. AB - A modified Asch (1951) conformity paradigm was used to study the impact of social influence on reality-monitoring decisions about new items. Subjects studied pictures of some objects and imagined others. In a later test phase, they judged whether items had been perceived in the study phase, had been imagined, or were new. Critically, for some items, the subjects were informed of a confederate's response before rendering a judgment. Although the confederate was always correct when they responded to old items, for new items, the confederate responded perceived, imagined, or new, or did not respond (baseline). In two experiments, we show that memory for new items was influenced by an erroneous response of the confederate. Social conformity was reduced by undermining the credibility of the confederate (Experiments 1A and 1B), and the confederate's influence was evident even after there was only a 20-min delay between study and test (Experiment 2), when the subjects were 87% accurate on new baseline items. These experiments reveal the power of social influence on reality-monitoring accuracy and confidence. PMID- 11407417 TI - Seeds aren't anchors. AB - Exposure to a few task-relevant numerical facts (seed facts) often improves subsequent numerical estimates. We performed two experiments to investigate the mechanism that produces these seeding effects. In Experiment 1, participants estimated national populations; in Experiment 2, they estimated between-city distances. In both, items were selected so that the actual value of the seed facts (SA) was, on average, below participants' initial estimates for those items (S1) and above the initial estimates for the transfer items (T1). Given this configuration, the anchoring position predicts that the postseeding transfer estimates should be greater than the preseeding transfer estimates (T2 > T1), whereas the feedback/induction position predicts the opposite (T2 < T1). In both experiments, the latter pattern of results emerged, supporting the conclusion that seeds aren't anchors. PMID- 11407418 TI - Memory for detail in item versus associative recognition. AB - Some studies have shown that, although repetition increases the familiarity of a stimulus, it does not improve memory for its details. Because memory for associative information is thought to require memory for the details of study presentation, the effects of repetition on associative recognition were examined in the present study. The pattern of results was similar to that found for the recognition of item details: Repetition increased the familiarity of the individual items within each pair to a greater extent than it improved memory for their specific pairings. PMID- 11407419 TI - False recall and false recognition induced by presentation of associated words: effects of retention interval and level of processing. AB - The effects of retention interval and level of processing on false recall and false recognition of associates were examined. False recall and false recognition were induced by presenting subjects with words closely associated with a non studied word. Both level of processing and retention interval affected false recall (Experiment 1) and false recognition (Experiment 2) in the same direction with which they affected accurate recall and accurate recognition. That is, semantically processed lists exhibited higher levels of later false recall and false recognition than did superficially processed lists. Furthermore, a decline in false recall and false recognition occurred across retention intervals of 0, 2, and 7 days. However, the decline in false recall and false recognition was less pronounced than the decline in accurate recall and accurate recognition. Results are consistent with source monitoring and fuzzy trace explanations of false recall and false recognition. PMID- 11407420 TI - Impact of encoding depth on awareness of perceptual effects in recognition memory. AB - Pictorial stimuli are more likely to be recognized if they are the same size, rather than a different size, at study and at test. This size congruency effect was replicated in two experiments in which the encoding variables were respectively undivided versus divided attention and level of processing. In terms of performance, these variables influenced recognition and did not influence size congruency effects. But in terms of awareness, measured by remember and know responses, these variables did influence size congruency effects. With undivided attention and with a deep level of processing, size congruency effects occurred only in remembering. With divided attention and with a shallow level of processing, size congruency effects occurred only in knowing. The results show that effects that occur in remembering may also occur independently in knowing. They support theories in which remembering and knowing reflect different memory processes or systems. They do not support the theory that remembering and knowing reflect differences in trace strength. PMID- 11407421 TI - Comparing viewer and array mental rotations in different planes. AB - Participants imagined rotating either themselves or an array of objects that surrounded them. Their task was to report on the egocentric position of an item in the array following the imagined rotation. The dependent measures were response latency and number of errors committed. Past research has shown that self-rotation is easier than array rotation. However, we found that imagined egocentric rotations were as difficult to imagine as rotations of the environment when people performed imagined rotations in the midsagittal or coronal plane. The advantages of imagined self-rotations are specific to mental rotations performed in the transverse plane. PMID- 11407422 TI - Susceptibility to semantic illusions: an individual-differences perspective. AB - When asked How many animals of each kind did Moses take on the ark?, people frequently respond "two" even though they know it was Noah, not Moses, who took animals on the ark. We replicate previous research by showing that susceptibility to semantic illusions is influenced by the semantic relatedness of both the impostor word and the surrounding context. However, we also show that the two text manipulations make independent contributions to semantic illusions, and we propose two individual-differences mechanisms that might underlie these two effects. We propose that the ability to resist the lure of a semantically related impostor word is related to the individual's skill at accessing and reasoning about knowledge from long-term memory. And we propose that the ability to resist the lure of the surrounding sentential context is related to the individual's capacity to simultaneously process and store information in working memory. PMID- 11407423 TI - The representations of the arithmetic operations include functional relationships. AB - Current theories of mathematical problem solving propose that people select a mathematical operation as the solution to a problem on the basis of a structure mapping between their problem representation and the representation of the mathematical operations. The structure-mapping hypothesis requires that the problem and the mathematical representations contain analogous relations. Past research has demonstrated that the problem representation consists of functional relationships, or principles. The present study tested whether people represent analogous principles for each arithmetic operation (i.e., addition, subtraction, multiplication, and division). For each operation, college (Experiments 1 and 2) and 8th grade (Experiment 2) participants were asked to rate the degree to which a set of completed problems was a good attempt at the operation. The pattern of presented answers either violated one of four principles or did not violate any principles. The distance of the presented answers from the correct answers was independently manipulated. Consistent with the hypothesis that people represent the principles, (1) violations of the principles were rated as poorer attempts at the operation, (2) operations that are learned first (e.g., addition) had more extensive principle representations than did operations learned later (multiplication), and (3) principles that are more frequently in evidence developed more quickly. In Experiment 3, college participants rated the degree to which statements were indicative of each operation. The statements were either consistent or inconsistent with one of two principles. The participants' ratings showed that operations with longer developmental histories had strong principle representations. The implications for a structure-mapping approach to mathematical problem solving are discussed. PMID- 11407424 TI - Misguided multiplication: creating false memories with numbers rather than words. AB - We built Deese (1959)/Roediger and McDermott (1995) (DRM) false memory lists composed of multiplication problems rather than words. Half these lists contained table-related, near neighbors (e.g., 3 x 7 = ??, 3 x 9 = ??) of a missing multiplication answer lure (e.g., 24). The other half contained problems unrelated to the lure (e.g., 5 x 5 = ??, 11 x 3 = ??). Participants solved each problem in a single list and then took immediate recognition (Experiment 1) or recall and then recognition tests (Experiment 2) for the answers. Many people misremembered that the lure was an answer to a study-phase problem, but only when solving the study list that contained the lure's neighbors. False memory was also greater for some list-lure combinations than others, as seen previously with words. We have thus demonstrated that numbers can also produce false memory, and we use the mental math and DRM task literatures to explain these results. PMID- 11407425 TI - Individual differences in working memory capacity and enumeration. AB - Two experiments are reported in which subjects performed working memory and enumeration tasks. In the first experiment, subjects scoring low on the working memory task also performed poorly on the attention-demanding "counting" portion of the enumeration task. Yet no span differences were found for the non-attention demanding "subitizing" portion. In Experiment 2, conjunctive and disjunctive distractors were added to the enumeration task. Although both high and low working memory span subjects were adversely affected by the addition of conjunctive distractors, the effect was much greater for the low-span subjects. Implications from these findings are that differences in working memory capacity correspond to differences in capability for controlled attention. PMID- 11407426 TI - The effect of lexical, pragmatic, and morphological violations on reading time and deviance ratings of English and German sentences. AB - Effects on reading time and deviance ratings of word-choice violations were studied at the surface code, propositional textbase, and situation model levels of representation in German and English sentences. Lexical violations (propositional textbase level) such as The housewife massaged the bread dough were rated as more deviant from normal language than were pragmatic violations (situation model level) such as The police officer shot the parking violator, although the pragmatic violations took longer to comprehend. The addition of a morphological (surface code) violation through the wrong form of an article (e.g., a ugly car) decreased reading time but only in cases where that material was deeply processed and where the morphology carried substantial syntactic information (e.g., in German, the definite article carries case, gender, and number information). The results thus confirmed the operation of comprehension strategies at multiple levels of representation and identified some cross linguistic generality in comprehension and some language differences. PMID- 11407427 TI - The gender marking effect in spoken word recognition: the case of bilinguals. AB - It has been known for some time that the recognition of a noun is affected by the gender marking, such as masculine or feminine, that is carried by a preceding word. In this study, we used auditory naming to examine how early and late English-French bilinguals react to gender marking when processing French. The early bilinguals showed clear facilitation and inhibition effects, but the late bilinguals were totally insensitive to gender marking, whether congruent or incongruent. The results are discussed in terms of current accounts of gender processing as well as age of acquisition and regular use of the gender-marking language. PMID- 11407428 TI - Eye movements during the production of nouns and pronouns. AB - Earlier research has established that speakers usually fixate the objects they name and that the viewing time for an object depends on the time necessary for object recognition and for the retrieval of its name. In three experiments, speakers produced pronouns and noun phrases to refer to new objects and to objects already known. Speakers looked less frequently and for shorter periods at the objects to be named when they had very recently seen or heard of these objects than when the objects were new. Looking rates were higher and viewing times longer in preparation of noun phrases than in preparation of pronouns. If it is assumed that there is a close relationship between eye gaze and visual attention, these results reveal (1) that speakers allocate less visual attention to given objects than to new ones and (2) that they allocate visual attention both less often and for shorter periods to objects they will refer to by a pronoun than to objects they will name in a full noun phrase. The experiments suggest that linguistic processing benefits, directly or indirectly, from allocation of visual attention to the referent object. PMID- 11407429 TI - Pseudohomophones and word recognition. AB - Pseudohomophones play an important role in visual word recognition research, but they are not often themselves the object of experimental inquiry. In Experiment 1, we explored whether the status of body rime relations in pseudohomophones whether their body rime relations exist in actual words-predicts the likelihood of word pronunciations to pseudohomophone spellings. In Experiment 2, we tested whether extant body rime relations modulate performance to pseudohomophones, and their context effect on word trials, in a lexical decision task. Extant body rime relations increase the likelihood that a pseudohomophone will be given a word pronunciation, and they produce slower and more error prone performance to pseudohomophones and words in lexical decision. PMID- 11407430 TI - When SOFA primes TOUCH: interdependence of spelling, sound, and meaning in "semantically mediated" phonological priming. AB - Three experiments test for semantically mediated priming of a word's phonology (e.g., sofa, an associate of couch, primes naming performance to touch). In the first two experiments, words that were body-rime-inconsistent (compare touch to couch) were used as naming targets. In the third experiment, words that were body rime-consistent were also used (i.e., sofa primed pouch). Low-frequency inconsistent words yield a high rate of pronunciation errors when they were primed by indirectly related words, such as sofa, in both a standard naming task and a speeded naming task. High-frequency inconsistent words yielded slower naming times when they were primed by indirectly related words in a speeded naming task, but consistent words showed no significant effects of the primes. The results suggest that the relationship between semantics and phonology plays an important, early role in word perception. PMID- 11407431 TI - Memory for unidentified items: evidence for the use of letter information in familiarity processes. AB - Participants can give accurate recognition judgments to word fragments that they are unable to complete. In three experiments, the generality of this finding was examined across tasks. Accurate memory judgments in the absence of identification were obtained in item recognition and judgments of presentation frequency but not in associative recognition or list discrimination. The former two tasks are thought to involve the use of familiarity; the latter two are thought to rely on recollection. The present results are consistent with the claim that recognition without identification reflects familiarity processes. PMID- 11407432 TI - Characteristics of adolescents' sexual partners and their association with use of condoms and other contraceptive methods. AB - CONTEXT: While a number of studies have examined the association between individuals' characteristics and their contraceptive use, few studies have examined the influence of partners' characteristics on individuals' contraceptive use. METHODS: Using nationally representative data from the National Longitudinal Study of Adolescent Health, multiple logistic analyses were conducted to identify associations between the demographic characteristics of adolescents' heterosexual partners and adolescents' use of condoms or other contraceptive methods. RESULTS: The partners of white and black adolescents were likely to be similar to them, while the partners of Latino adolescents and of adolescents of "other" race or ethnicity were more likely to be of a different racial or ethnic group. Differences in age between adolescents and their partners were notable in all racial and ethnic groups. As adolescents age, the characteristics of their partners become more heterogeneous. The less similar adolescents and their partners are to one another--whether because of a difference in age, grade or school--the less likely adolescents are to use condoms and other contraceptive methods. CONCLUSIONS: Many adolescents have relationships with partners whose characteristics differ from theirs and with whom they are less likely to use condoms or other contraceptive methods. This behavior is more common as adolescents grow older. To provide appropriate counseling, sexuality educators and family planning providers need to consider the ways in which adolescents' relationships change as they age and discuss with them the dynamics of relationships involving partners who differ in age or other characteristics. PMID- 11407433 TI - Risk factors for HIV and other sexually transmitted diseases among adolescents in St. Petersburg, Russia. AB - CONTEXT: Over the past several years, there have been sharp increases in the prevalence of HIV and other sexually transmitted diseases (STDs) among young people in Russia. Very little is known about Russian adolescents' behaviors and attitudes that might influence their risk of acquiring these infections. METHODS: A 1995 survey of 533 students aged 15-17 attending eight St Petersburg high schools assessed their sexual risk practices, AIDS-specific attitudes and beliefs, sexual relationship patterns and preferences, and social characteristics. RESULTS: Overall, 39% of students were sexually experienced, and these young people had had, on average, 3.4 sexual partners. Only 29% of sexually experienced students said they consistently used condoms, and 29% said they never did. Unprotected vaginal intercourse was the predominant and preferred sexual practice; it also was the practice that most often occurred with students' last sexual partner. In all, 28% of students defined "safer sex" as condom use. Many young people believed that AIDS is a threat only to members of particular "risk groups"; relatively few believed that they could get AIDS (17%) or said that AIDS information had influenced their sexual behavior (29% of those who were sexually experienced). Females were more likely than males to prefer having an exclusive partner, and males were more likely to prefer having casual partners. CONCLUSIONS: Educational and behavioral interventions are urgently needed to help young people in Russia avoid HIV and other STDs. Risk and social characteristics identified in this study can help to guide the development and tailoring of risk reduction interventions. PMID- 11407434 TI - Family planning clinic services in the United States: patterns and trends in the late 1990s. AB - CONTEXT: Publicly funded family planning clinics are a vital source of contraceptive and reproductive health care for millions of U.S. women. It is important periodically to assess the number and type of clinics and the number of contraceptive clients they serve. METHODS: Service data were requested for agencies and clinics providing publicly funded family planning services in the United States in 1997. The numbers of agencies, clinics and female contraceptive clients were tabulated according to various characteristics and were compared with similar data for 1994. Finally, county data were tabulated according to the presence of family planning clinics and private physicians likely to provide family planning care and according to the number of female contraceptive clients served compared with the number of women needing publicly funded care. RESULTS: In 1997, 3,117 agencies offered publicly funded contraceptive services at 7,206 clinic sites. Forty percent of clinics were run by health departments, 21% by community health centers, 13% by Planned Parenthood affiliates and 26% by hospitals or other agencies. Overall, 59% of clinics received Title X funding. Agencies operated an average of 2.3 clinics, and clinics served an average of 910 contraceptive clients per year. Altogether, clinics provided contraceptive services to 6.6 million women-approximately two of every five women estimated to need publicly funded contraceptive care. The total number of providers and the total number of women served remained stable between 1994 and 1997; at the local level, however, clinic turnover was high. Some 85% of all US counties had one or more publicly funded family planning clinics; 36% had one or more clinics, but no private obstetrician-gynecologist. CONCLUSIONS: Publicly funded family planning clinics are distributed widely throughout the United States and continue to provide contraceptive care to millions of US women. Clinics are sometimes the only source of specialized family planning care available to women in rural counties. However, the high rate of clinic tumover and the lack of significant growth in clinic numbers suggest that limited funding and rising costs have hindered the further expansion and outreach of the clinic network to new geographic areas and hard-to-reach populations. PMID- 11407435 TI - Knowledge of and attitudes toward the pill: results of a national survey in Japan. AB - CONTEXT: After decades of debate, the oral contraceptive pill was legalized in Japan in June 1999. Because the pill had been unavailable up until then, little is known about the public's knowledge about, attitudes toward and intentions to use the pill. METHODS: In a nationwide probability sample, 630 women and men were interviewed in their homes in March 1999. Respondents were asked a range of questions to gauge their knowledge about and attitudes toward the pill. RESULTS: Most respondents (79%) could identify both the name and purpose of the birth control pill. Roughly the same proportion of respondents held an overall positive impression (44%) as held a negative impression (42%) of the pill, and 14% were undecided about the method. The pill's high level of effectiveness in pregnancy prevention was the most common reason for having a favorable impression of it (47%). Women were more concerned about side effects than were men, and they also knew more about the pill's potential side effects than did men. Only 12% of respondents said they intended to use the pill if it were approved. Roughly one quarter (23-26%) did not correctly identify the pill's inability to protect against HIV and other sexually transmitted diseases (STDs). CONCLUSIONS: Legalization of oral contraceptives in Japan has led to an urgent need to educate both men and women on the inability of the pill to protect against STDs, including HIV Policymakers and providers need to recognize the importance of encouraging dual method use in Japan. PMID- 11407436 TI - The media, public opinion and population assistance: establishing the link. PMID- 11407437 TI - New target group for HIV prevention. PMID- 11407438 TI - Epidemiology of severe insomnia and its consequences in Germany. AB - This is the first nation-wide face-to-face survey on the prevalence of well defined severe insomnia and its impact on quality of life in the general population of Germany. The survey was part of an international epidemiological study, which was also conducted in Belgium, Great Britain, Ireland and Sweden. A representative sample of 1913 adults aged 18 years and over were interviewed in all parts of Germany according to the quota method. Subjects with symptomatic insomnia were identified using an algorithm compatible with the principal criteria for severe insomnia defined in the fourth revision of the Diagnostic and Statistical Manual of Mental Diseases (DSM-IV). Subjects provided data on quality of life using the Short Form 36 Health Survey (SF-36) questionnaire and on health care consumption. Prevalence of severe insomnia in Germany was found to be 4%, which was lower than in other European countries (6-22%). Severe insomnia was more prevalent among women, the unemployed, those living alone after divorce or separation, and those in large cities, but not more frequently in the elderly (aged 65 years and over). The majority of subjects had chronic complaints, with 74 % of them suffering from severe sleep problems for over a year's duration (average 56+/-23 months). Consultations with general physicians, medication usage, medical tests and hospitalisation were greater among severe insomniacs compared to subjects who had no sleep complaints. The question regarding overall appreciation of quality of life was rated as bad in 22% and good in 28% of severe insomniacs compared to 3% (bad) and 68 % (good) in subjects with no sleep complaints. Despite this, only 55% of severe insomniacs had ever discussed their sleep problem with a doctor and the proportion who consulted their doctor specifically regarding sleep problems in the previous 12 months was even lower (36%). The vast majority (73%) was not taking hypnotic or sedative medications. In conclusion, insomnia, even when severe, is a common and a chronic complaint in Germany. This trial suggests that while, on the one hand, sleep disorders have a significant impact on patient's quality of life and consumption of health care, it is, on the other hand, a condition that is poorly recognised and for which patients are, for unknown reasons, reluctant to seek treatment. PMID- 11407439 TI - Association between TaqI A dopamine D2 receptor polymorphism and therapeutic response to bromperidol: a preliminary report. AB - The relationship between TaqI A dopamine D2 receptor (DRD2) polymorphism and therapeutic response to bromperidol, a selective dopamine antagonist, was investigated in 30 acutely exacerbated schizophrenic inpatients. Patients were treated with bromperidol 6-18 mg/day for 3 weeks. Clinical symptoms were evaluated by the Brief Psychiatric Rating Scale (BPRS) before and 3 weeks after the treatment. The TaqI A genotypes were determined with the PCR method. There was no significant difference in the percentage improvement of total BPRS or 5 subgrouped symptoms (positive, negative, anxiety-depression, excitement and cognitive symptoms) after the 3-week treatment between the patients with A1 alleles (n=18) and those with no A1 allele (n=12). Although the present study is preliminary, it is suggested that the TaqI A DRD2 polymorphism is not associated with therapeutic response to bromperidol in schizophrenic patients. PMID- 11407440 TI - Remitted schizophrenia-spectrum patients with spared working memory show information processing abnormalities. AB - Working memory and information processing abnormalities are often reported in schizophrenia. The aim of this study was to examine visual backward masking (BM) functions in remitted schizophrenia-spectrum patients with spared working memory functions. Seventy-two patients with DSM-IV schizophrenia-spectrum disorders were screened using the Wisconsin Card Sorting Test (WCST) and the digit span forward/backward tasks. Patients with spared WCST and digit span performances were selected and administered a spatial working memory test and two BM procedures (target identification and location). The schizophrenia-spectrum group with spared WCST and digit span performances included individuals with schizophreniform disorder (N=11), schizophrenia (N=2), and schizoaffective disorder (N=2). These patients were clinically remitted and demonstrated spared IQ, normal spatial working memory, and relatively high psychosocial functioning. However, there was a significant impairment in the BM procedure, most prominently in the target location task and at short interstimulus intervals. These results suggest that the BM dysfunction is a trait marker of schizophrenia-spectrum disorders and may be present in the absence of working memory abnormalities. PMID- 11407441 TI - Hypofrontality in neuroleptic-naive schizophrenic patients during the Wisconsin Card Sorting Test--a fMRI study. AB - Functional neuroimaging findings of "hypofrontality" in schizophrenic patients - as tested with the Wisconsin Card Sorting Test (WCST) - are still controversial, mainly due to methodological aspects and the heterogeneity of the patient samples. To measure WCST specific and reproducible reduced cerebral activations in schizophrenic patients, we revised the study design and patient recruitment, respectively. For this purpose, we used an adequate active control task instead of an undefined rest condition to determine exclusively WCST specific cerebral activations. In addition, we focused on the investigation of modified activations between a selected group of neuroleptic-naive schizophrenic patients and carefully matched healthy controls by means of functional magnetic resonance imaging. The results indicate that neuroleptic-naive schizophrenic patients show reduced activations in the right frontal and left temporal lobe, as well as in the left cerebellum. By utilizing an active control task all unwanted activations are suppressed. Furthermore the influence of different task performances is reduced. The findings are in line with previous PET and SPECT studies and confirm the "hypofrontality" hypothesis. The findings suggest that "hypofrontality" is not caused by neuroleptic medication. PMID- 11407442 TI - Effectiveness of targeted intervention and maintenance pharmacotherapy in conjunction with family intervention in schizophrenia. AB - A sample of 85 patients with schizophrenia, of whom 34 later dropped out, received randomised treatment. There were no significant differences between treatment-takers and drop-outs in the variables assessed. Patients received either standard-dose maintenance neuroleptic treatment or targeted maintenance pharmacotherapy and all patients received behavioural family therapy. Measures of psychopathology, social adjustment, side-effects, family burden, and expressed emotion were assessed at baseline and then periodically over an 18-month period. The study was designed to compare the two alternative pharmacological maintenance approaches, each of them supported by psychosocial intervention. Any evaluation of the impact of behavioural family treatment on relapse rates and other outcome criteria is exclusively descriptive. A significantly higher rate of relapse was observed at 18 months in patients randomised to targeted treatment compared to those randomised to standard-dose treatment (35% vs 4%). Although patients assigned to the targeted maintenance group received significantly lower mean doses of neuroleptics, there were no significant differences between the two groups with regard to side-effects, global measures of social function, and overall psychopathology. Family burden was higher in the targeted-treatment group at six months, but did not differ at the one-year and eighteen-month time points. However, both groups improved significantly from baseline to 12 or 18 months in almost all variables assessed. Thus, the behavioural family approach did not compensate for the problems associated with the targeted medication strategy. PMID- 11407443 TI - Increased prevalence of obesity in narcoleptic patients and relatives. AB - Increased Body Mass Indices (BMIs), increased prevalences of non insulin dependent diabetes and sleep apnoe syndrome have been reported to be associated with narcolepsy. Our objective was to explore and possibly confirm the association of narcolepsy and increased BMI. In addition, we addressed the question whether increased BMIs also occur in relatives of narcoleptic patients. Together with narcolepsy-related clinical parameters we measured body weight and height of 132 narcoleptic patients who agreed to participate in our narcolepsy research program. In addition, 52 first degree relatives of 22 index patients, mostly from multiplex families, were included in the study. Data were compared to published general population surveys, recently conducted in Germany and Switzerland as well as to collective of 104 psychiatric inpatients. Narcoleptic patients had significantly increased BMIs in comparison to general populations or psychiatric controls. BMIs of first degree relatives were lower than those of index patients but significantly higher than those found in the general population. BMIs were not related to symptom severity or to medication status. Thus, the elevated BMIs appeared not to be secondary to behavioral consequences of narcolepsy but may reflect a trait at least partially common to index patients and relatives. PMID- 11407444 TI - Different immune patterns in melancholic and non-melancholic major depression. AB - The search for immune patterns in major depression has thus far resulted in ambiguous findings, probably because patient samples are psychiatrically heterogeneous. We therefore focused on a detailed classification of subtypes of major depression, comparing patients with melancholic and non-melancholic major depression. Inpatients suffering from acute major depression were diagnosed and subclassified according to DSM IV criteria. Cell counts were determined by FACS analysis and morphology. Cytokine production (IL-2, IFN-gamma, IL-10) upon mitogen stimulation was measured by ELISA in a whole blood assay. Non-melancholic patients showed increased counts of leukocytes, lymphocytes and NK-cells in the acute stage of disease and after two and four weeks of treatment. Their lymphokine production was unchanged compared to that of healthy controls. Melancholic patients on the other hand demonstrated normal cell counts but a decreased production of IL-2, IFN-gamma and IL-10 during the acute stage of disease followed by a normalization with clinical improvement. Melancholic and non-melancholic patients showed different immune patterns. Classifying melancholic and non-melancholic patients is helpful towards the identification of immune characteristics typical for these diseases. PMID- 11407445 TI - Barotrauma of the ears and sinuses after scuba diving. AB - The pathophysiology, differential diagnosis, and currently available management of barotrauma affecting the ears and sinuses after scuba diving are reviewed, along with medical standards for resuming scuba diving after barotrauma has resolved. PMID- 11407446 TI - Expression of myeloperoxidase and cochlear dysfunction in the lipopolysaccharide treated guinea pig. AB - In this study, the effect of endotoxin on the guinea pig cochlea was examined by electrophysiology and immunohistochemistry. Bacterial lipopolysaccharide (LPS) was injected into the middle ear transtympanically. Electrocochleograms were measured immediately and 48 h after the injection with an electrode inserted into the facial canal. After the electrophysiological measurement, the animals were killed with an intracardiac perfusion of fixative and the temporal bones were removed and processed for immunohistochemistry with anti-myeloperoxidase (MPO) antibody. MPO could be detected after 48 h in the lateral wall and the organ of Corti. After injection of LPS, the threshold of the compound action potential worsened significantly at 48 h in the LPS group. These results suggest that MPO and reactive oxygen species are involved in cochlea dysfunction under inflammatory conditions. PMID- 11407447 TI - A threshold-like measure for the assessment of olfactory sensitivity: the "random" procedure. AB - Many tests of olfactory dysfunction are either too complex, too expensive, or too time-consuming to be of use in routine clinical testing. Thus, the present multicenter study was undertaken to investigate a new approach, the so-called "random" test. In this test different concentrations of citronellal and phenyl ethyl alcohol are applied according to a pre-established order; patients are asked to identify the odor if possible. The test score is the sum of correctly identified odors. Test administration takes about 10 min. Two studies were performed. Basic characteristics of the test were explored in experiment 1 in 176 healthy subjects (76 male, 100 female; age 12-85 years, mean age 30 years), namely test-retest reliability, correlation with other measures of olfactory sensitivity, and sensitivity of the test to differences in age and gender. In the second experiment the test was tried in 97 patients (45 male, 52 female; age 19 78 years, mean age 47 years) in a clinical environment to investigate its usefulness in diagnosing olfactory loss. The "random"-test was found (1) to exhibit a test-retest reliability similar to that reported for established measures of olfactory function (r = 0.71; P < 0.001), (2) to correlate with other measures of olfactory sensitivity (0.82 > r > 0.60; P < 0.001), (3) to differentiate between expected differences in olfactory sensitivity in relation to gender (t > 2.602, P < 0.011), and (4) to discriminate between different degrees of olfactory loss (F > 36.6, P < 0.001). Based on these data, and the fact that the new test requires little time and is easy to use, this approach can be expected to suit clinical needs. PMID- 11407448 TI - Voice restoration after circumferential pharyngolaryngectomy with free jejunum repair. AB - Speech restoration after circumferential pharyngolaryngectomy with free jejunal repair for advanced tumors of the hypopharyngo-esophageal tract remains a difficult problem to solve. We report here the results of secondary voice restoration in six patients who received a Provox 2 type prosthesis and intensive speech therapy after circumferential pharyngolaryngectomy with free jejunum repair. No patient had operative or post-operative complications due to insertion of the prosthesis. No patient had to have the prosthesis removed during the follow-up (8 to 14 months). Analysis of some acoustic parameters of voice (fundamental frequency, waveform perturbations) and qualitative characteristics of speech (intelligibility, pleasantness and acceptability) demonstrated that all the patients were able to produce satisfactory speech after tracheojejunum puncture and speech therapy and were satisfied with their own ability to communicate. Our results are reassuring and we therefore advise that in patients undergoing free jejunum flap reconstruction of the hypopharyngo-esophageal tract voice restoration should be attempted by placing a voice prosthesis through a secondary tracheo-esophageal puncture and providing intensive speech training. PMID- 11407449 TI - The dilemma of follow-up in head and neck cancer patients. AB - The aims of tumor follow-up in head and neck cancer patients are (1) evaluation of therapeutic efficacy, (2) management of impairments, (3) detection of new tumor manifestations, and (4) psychosocial care. In general standardized 5-year protocols are used for all such patients. However, it is questionable whether a rigid follow-up schedule is optimal for a very heterogeneous tumor population. Therefore 603 patients with sqamous cell carcinoma of the oral cavity, pharynx or larynx, or with cervical metastasis from an unknown primary site (CUP syndrome), who had been diagnosed and treated curatively by an operation with or without radiotherapy (n = 523) or just by radio(chemo)therapy (n = 80) between 1985 and 1994, and who had been followed-up regularly according to a standardized plan, were worked-up retrospectively. Data were evaluated for the manifestation and prognosis of curable new tumor manifestations as well as for tumor-specific factors likely to select groups which should be followed more or less intensively. Within a 5-year follow-up period new tumor growth was detected in 152/603 (25%) patients: 79 local and 31 regional recurrences, 18 systemic metastases and 24 second primary cancers. Where follow-up was extended beyond the 5th year, 168/603 (28%) patients presented a new tumor manifestation. One hundred and sixteen of the 152 (28%) patients had another operation with or without radiotherapy or had radio(chemo)therapy alone. So far 18/116 (14%) patients have survived their new tumor manifestation for more than 5 years and 30/116 for more than 2 years. Tumor-specific data on the initial tumors (T stage, N stage, site) did not indicate the risk of a new tumor manifestation, but 87% of patients who survived their new tumor manifestation for more than 2 years initially had T1 or T2 tumors and only 30% initially had N+ necks. Occurrence of distant metastasis or a second primary outside the head and neck region limited survival to < or = 2 years after detection. In terms of survival, follow-up efforts should therefore concentrate on detection of locoregional recurrence, particularly if an option for further curative local therapy exists. The limited success of detection of new tumor manifestations in terms of survival does not justify a reduction in tumor-follow-up examinations, since the benefit of the other efforts cannot be determined from survival figures. PMID- 11407450 TI - Pharyngo-esophageal diverticulum arising from Laimer's triangle. AB - Among pharyngo-esophageal diverticula the most frequent type is Zenker's diverticulum, which originates above the cricopharyngeal muscle and below the thyropharyngeal muscle. Laimer's diverticulum, located below the cricopharyngeal muscle, is a much rarer type. We report a case of a 54-year-old woman with a 3 year history of pharyngeal discomfort that was diagnosed as Laimer's diverticulum. To the best of our knowledge, this is only the third such reported case. PMID- 11407451 TI - Positive surgical margins in cancer of the larynx. AB - In order to determine what should be done for laryngeal cancer patients when surgical margins are positive, and to evaluate their prognosis, a retrospective review of 21 laryngeal cancer patients with positive surgical margins out of 714 surgically treated cases (2.9%) was carried out. Nineteen patients were treated with postoperative radiation therapy. Two patients who had had endolaryngeal partial laryngectomy were treated with vertical partial laryngectomy. Two patients were lost to follow-up. Ten patients (10/19; 53%) were recurrence-free. Four patients had local, two had regional, and two had locoregional recurrences. Only one patient with a local recurrence could be salvaged with total laryngectomy and is disease-free. One patient developed liver metastasis. Nineteen patients had a mean and median disease-free survival of 48 and 36 months, respectively. Nine out of fourteen patients (64%) treated curatively were recurrence-free. The patients with positive margins developed significantly more locoregional recurrences than those with free margins (P < 0.05). We conclude that surgical margins must be checked peroperatively with frozen sections to make sure that they are free. The margins of every laryngectomy specimen must be diligently examined. If positive, re-excision, postoperative radiotherapy and chemotherapy are treatment alternatives. They should not just be managed with close follow-up. However, whatever treatment is applied, the prognosis for patients with positive margins is significantly worse than for those with free margins. PMID- 11407452 TI - Resection of the sternocleidomastoid muscle during radical neck dissection. AB - BACKGROUND: Surgical therapy of lymph node metastasis is based on accessibility for en bloc resection. First described as "radical neck dissection", this original approach has since undergone various modifications. This has produced controversy about the particular indications for the individual techniques. The aim of this study was to evaluate whether intraoperative macroscopic inspection of the sternocleidomastoid muscle (SCM) in regard to tumor infiltration is sufficient to decide about muscle resection and whether there are prognostic differences between patients undergoing radical-versus modified radical (selective) neck dissection. MATERIALS AND METHODS: In a retrospective study, data on the surgical treatment of cervical lymph nodes and survival rates from 438 patients with head and neck malignancies managed in our department between 1988 and 1994 were analyzed in 1994 and again in 1999. RESULTS: 337 patients (76.9%) underwent unilateral or bilateral selective neck dissection. In 101 patients (23.1%) a radical neck dissection was performed and the SCM was completely resected. Analysis of these cases showed intraoperative macroscopic tumor invasion of the SCM in 12 patients (11.9%), which could be confirmed histologically. In the remaining 89 cases (88.1%), a macroscopically intact muscle was resected; in none of these cases did histopathological examination show tumor infiltration of the SCM. Analysis of radically or selectively neck dissected stage III or IV patients with oral cavity, oropharyngeal, hypopharyngeal or laryngeal carcinomas did not show statistical differences in 2 , 5- and 10-year survival (54.8%, 23.7%, 18.7% versus 62.6%, 25.6%, 21.8%, respectively). CONCLUSIONS: (1) Intraoperative inspection of the SCM constitutes a valid parameter for deciding whether tumor infiltration is present or not. (2) There were no prognostic differences (2-year, 5-year and 10-year-survival) between stage III and IV patients with oral cavity, oropharyngeal, hypopharyngeal and laryngeal carcinomas treated by either radical or selective neck dissection. PMID- 11407453 TI - Wegener's granulomatosis presenting with bilateral facial nerve palsy. AB - Primary manifestation of Wegener's granulomatosis in the mucosa of the middle ear is rather rare, and has been reported as presenting with serous otitis media, chronic otitis media, sensorineural hearing loss, and, in rare instances, unilateral facial palsy. Bilateral facial palsy has never been reported. This last fact constitutes the interest in our report of a 23-year-old female patient who presented with symptoms of recurrent bilateral otitis media, eventually developing sensorineural hearing loss and bilateral facial palsy. Soon thereafter neurological symptoms appeared and lung extension was noted. Histological examination of repeated biopsies taken from the nasal and middle ear mucosa was not conclusive for the suspected disease, and c-ANCA titers were also initially repeatedly negative. Eventually, positive lung biopsy and elevated c-ANCA titers when the patient had developed pulmonary granulomas confirmed the diagnosis of Wegener's granulomatosis. Mastoid surgery with facial nerve decompression of the most severely afflicted side did not result in the recovery of facial nerve function. Medical therapy with corticosteroids and cyclophosphamide improved the clinical picture but were ineffective in improving the bilateral sensorineural hearing loss and the facial paralysis on the operated side. We would contribute to the literature a unique case of bilateral facial nerve palsy due to Wegener's granulomatosis. PMID- 11407454 TI - Langerhans' cell granulomatosis in an adult: a 22-year follow up. AB - A 57-year-old patient with Langerhans' cell granulomatosis (LCG) is presented. During 22 years of treatment, there were five relapsing infiltrations at different sites of the skull bones, which were treated by surgery, local radiotherapy, and chemotherapy. During the last relapse, the right temporal bone was infiltrated by granuloma and the petrous bone was destroyed with an intimate spread to the internal carotid artery. After two palliative surgical resections and ineffective radiotherapy, 12 cycles of chemotherapy (vinblastine plus prednisolone) were applied and a clinical remission of the disease was achieved. Special attention is paid to the complexity of diagnosis and choice of therapy. It is concluded that the behavior of LCG may change with time, and assume an aggressive form of the disease. Chemotherapy is the treatment of choice for this type of multifocal malignant form of LCG. Prognosis of the disease is unpredictable. PMID- 11407455 TI - Domestic violence and women's mental health in Japan. AB - There are positive changes in both the social and legal understanding of domestic violence in Japan. However, the scope of the problem has not been investigated in depth and described in the Japanese nursing literature. This descriptive study of a random sample of 177 women investigated domestic violence and the relationship between domestic violence and the mental health of the victims. Sixty-seven per cent of the female respondents reported having experiences of physical, psychological and/or sexual abuse. Forty-seven per cent of the abused women achieved statistically significant General Health Questionnaire scores that indicated clinical depression or anxiety. The findings of this study will enable Japanese nurses to better assess and intervene on behalf of their patients. In addition, avenues for further nursing research are suggested. PMID- 11407456 TI - International nursing faculty exchange model: a Chile-USA case. AB - International faculty-exchange programmes can be cost-effective methods for faculty development and enhanced student learning. Despite the increasing interest in international nursing exchanges, the literature on faculty exchanges is limited. This article examines a case study of a successful international nursing faculty-exchange programme between a university in Chile and one in the United States of America (USA), based on a model for international faculty exchange. The model includes the components of pre-exchange planning, academic activities, socio-cultural events and evaluation of outcomes. Communication is at the centre of the model and ideally should be continuous, bilateral, flexible and technological. Examples from the case study illustrate teaching strategies and ongoing education of faculty. The article concludes with suggestions for successful international nursing faculty exchanges. PMID- 11407457 TI - Building international partnerships. AB - Health is a global concern. Although nursing is a global profession, most schools of nursing concentrate on teaching health exclusively within the context of their own nation. Sister-school partnerships that cross national boundaries are one way of extending the learning opportunities of faculties and students. An example of a 5-year partnership is described and analysed. PMID- 11407458 TI - The Supervised Methadone and Resettlement Team nurse: an effective approach with opiate-dependent, homeless people. AB - Homelessness and substance misuse have risen dramatically over the past 30 years in the UK. The role of the primary care nurse has been signalled as important in working with people who have drug and alcohol problems, and for improving the general health of homeless people. This article focuses on the role of the primary care nurse in a Supervised Methadone and Resettlement Team (SMART). The team works in central Bristol, in southwest England, with people who are homeless and using illegal opiates. The aim of this report is to provide descriptive information that demonstrates the value of the primary care nurse, working in a multiagency partnership, in dealing with the problems of this homeless population, many of whom have problems associated with illicit drug use. Client outcomes from a small sample of homeless persons are also described. PMID- 11407459 TI - Nurses moving across borders: 'brain drain' or freedom of movement? PMID- 11407460 TI - We're meeting, but are we organized? PMID- 11407461 TI - Warm hearts, inspiration and a sense of belonging. PMID- 11407462 TI - Sadako Ogata is first to receive new ICN award for health and human rights. International Council of Nurses. PMID- 11407463 TI - Advancing nursing in Zimbabwe. PMID- 11407464 TI - ICN project will help create a better world for young girls. International Council of Nurses. PMID- 11407465 TI - At issue: ethical recruitment. PMID- 11407466 TI - Career planning and development for nurses: the time has come. AB - Developments in how the nursing profession is perceived by nurses and by society, along with unparalleled changes in health care systems, have created an environment in which individual nurses must take control of their careers and futures. Educators, employers and professional organizations also have a key role to play in fostering the career planning and development of nurses, usually the largest employee group in most health care organizations. This article provides an overview of what career planning and development is and why it is important for nurses. A career planning and development model is described that provides nurses with a focused strategy to take greater responsibility for engaging in the ongoing planning process that is crucial throughout the major stages of their career. Finally, educators, employers and professional organizations are challenged to collaborate with individual nurses on career-development activities that will enable nurses to continue to provide high-quality care in ever-changing health care systems. PMID- 11407467 TI - Developing information literacy: a key to evidence-based nursing. AB - This report describes the evaluation of a curriculum-integrated programme designed to help students develop an awareness of the nursing literature, the skills to locate and retrieve it, and skills required in its evaluation; in other words'information literacy'. Positive changes in student performance on objective measures of information-literacy skills were revealed as well as a significant increase in the levels of confidence of the student in performing those skills. Students who had undertaken the information-literacy programme ('programme' students) performed better on a range of objective measures of information literacy, as well as reporting higher levels of confidence in these skills, than students who had not participated in the programme ('non-programme' students). Evaluation of this programme provides evidence of the potential usefulness of a curriculum-integrated approach for the development of information-literacy skills within nursing education. With these underlying skills, students will be better equipped to consolidate and extend their key information-literacy skills to include research appreciation and application. These are vital for effective lifelong learning and a prerequisite to evidence-based practice. PMID- 11407468 TI - Building an intercultural nursing terminology bank for the phenomenon, Violence, of the International Classification of Nursing Practice: a methodological perspective. AB - The aim of this article was to present the methodology and results of an investigation into the intercultural evolution of the nursing phenomenon- Violence--from the identification phase to the summative evaluation phase, in order to contribute to the evolution of the International Classification of Nursing Practice (ICNP). The organizing construct was concept analysis of a nursing phenomenon. The identification of a universal problem in psychiatric mental health nursing by a team of 10 international leaders in the speciality led from the phase of concept analysis to development of a postgraduate educational module. The results of the original analysis of the concept and findings from clinical and evaluation data resulted in further analysis and consequent synthesis of the concept. Results will be disseminated to the International Council of Nursing (ICN) for the International Classification of Nursing Practice (ICNP) to further the evolution of the phenomenon from a focus on the individual to that of the aggregate (social violence). The methodology pointed to a lack of transfer in the phenomenon from a focus on the individual to that of aggregates in the ICNP. There was also a paucity of intercultural components. The phenomena of Abuse and Aggression appear to have been used synonymously with Violence, without an explanation. The data, and the methods used for its collection and submission, will enhance the ICNP. PMID- 11407469 TI - Flow cytometric DNA and phenotype analysis in pathology. A meeting report of a symposium at the annual conference of the German Society of Pathology, Kiel, Germany, 6-9 June 2000. AB - This meeting report summarizes the presentations of three different groups that are active in the field of flow cytometry (FCM) in relation to diagnosing and classification of proliferative disorders. The report starts with the contribution from Regensburg about the developments in DNA FCM, the progression to dual parameter determinations, and combination of immunophenotyping in combination with DNA. In the second part, the use of FCM for the detection of isolated tumor cells in the peripheral blood from patients with prostate or breast cancer is discussed in a contribution from Munster. In the third part, from Heerlen, the use of multi-parameter FCM on formalin-fixed paraffin-embedded tissues from solid tumors is discussed as a new development and application in routine surgical pathology. PMID- 11407470 TI - Tenascin: a sensitive and specific diagnostic marker of minimal collagenous colitis. AB - Collagenous colitis is a rare cause of chronic watery diarrhea. In this condition, endoscopic findings are usually normal. Currently, the diagnosis relies on the histological presence of thick subepithelial bands of collagen deposits and an inflammatory infiltrate within the mucosa. However, these subepithelial bands may be developed only focally and may be too subtle to allow a definitive diagnosis upon routine hematoxylin and eosin (HE) and van Gieson's stainings. Recently, we and others were able to show a prominent staining of tenascin and type-VI collagen in the subepithelial band-like structures. In this study, we tested the diagnostic value of tenascin staining and type-VI collagen immunolocalization for the identification of collagenous colitis and compared it with conventional histology and histochemical detection of collagens. The analysis was based on 434 biopsy specimens of collagenous colitis, other forms of colitis, and normal mucosa. We were able to show that the immunohistochemical detection of increased amounts of tenascin, selectively in the subepithelial zone, is a specific test for collagenous colitis, with a sensitivity superior to conventional histological and histochemical detection, especially in minimal collagenous colitis (P<0.001). Of note, tenascin staining also allows the diagnosis of collagenous colitis in biopsies obtained only from the rectum and sigmoid colon, thus avoiding the need for colonoscopic investigations. Tenascin immunostaining is a simple and safe tool to complement conventional histological diagnostics in clinically and histopathologically unclear cases of diarrhea. PMID- 11407471 TI - Pancreatic ductal myofibroblasts. Proliferative patterns in various pathologic situations. AB - Myofibroblasts in the periacinar area of the pancreas have been demonstrated to mediate fibrogenesis in pancreatic fibrosis. However, only a few reports have described myofibroblasts in the pancreatic duct. To elucidate the presence of myofibroblasts in the pancreatic ductal wall, we performed an immunohistochemical study, using immunostains for both alpha-smooth muscle actin (alphaSMA) and desmin, and an electron microscopic study on surgically resected pancreatic specimens from 10, 23, 23, and 56 cases of focal pancreatitis (FP), chronic pancreatitis (CP), pancreatic carcinoma (PCa), and carcinoma of the papilla of Vater (VPCa), respectively. All cases showed localized stenosis of the main pancreatic duct by means of preoperative pancreatography. As controls, 20 autopsy cases were studied. alphaSMA-positive and desmin-negative cells existed in the ductal walls of controls and were revealed as myofibroblasts by means of electron microscopy. In six FPs, proliferation of myofibroblasts was observed at the stenotic portion. In VPCas, myofibroblasts mainly proliferated in the pancreatic ductal wall. In CPs and PCas, no myofibroblast proliferation was observed at the stenotic portion. The proliferation of myofibroblasts might occur as a wound healing process in FP, while acting against elevation of intraductal pressure in VPCa. In conclusion, proliferation of myofibroblasts plays an important role in ductal changes in various pathological situations. PMID- 11407472 TI - Establishment of cell lines from adenocarcinomas of the esophagus and gastric cardia growing in vivo and in vitro. AB - The aim of this study was to establish cell lines of adenocarcinomas of the gastro-esophageal junction(GEJ), which grow in vivo and in vitro. Primary esophageal and gastric cardia adenocarcinomas and corresponding lymph node metastases were xenografted subcutaneously to immunodeficient nude mice. In addition, tumor tissue was also used for in vitro culture. Xenografting of 70 primary adenocarcinomas and 17 metastases resulted in the initial growth of 22 and 6 tumors, respectively (total 32%). Upon retransplantation, six long-term xenografts [esophageal adenocarcinoma (OAC)P33X, OACP47X, OACP56X, OACP58X, OACP67X, OACP76X] from primary tumors and three (OACM2.1X, OACM30X, OACM53X) from metastases were obtained. In vitro culture attempts of 34 primary tumors and nine metastases resulted in the establishment of three (7%) permanent in vitro growing cell lines. From one patient, a cell line from the primary tumor (OACP4 C) and from a lymph node metastasis (OACM4.1 C) was established. The third cell line (OACM5.1 C) was also derived from a lymph node metastasis. The in vivo and in vitro cell lines were characterized using immunocytochemistry and microsatellite analysis to verify their epithelial and human tumor origin, respectively. PMID- 11407473 TI - Keloids and hypertrophic scars of Caucasians show distinctive morphologic and immunophenotypic profiles. AB - The aim of this study was to identify possible morpho-phenotypic differences between keloids (K) and hypertrophic scars (HS) in a Caucasian population. Young HS (< or =1 year of age) presented a high number of diffusely distributed spindle shaped cells (alpha-smooth-muscle actin+ and fibronectin+). Fully developed HS (> 1 year of age and <3 years of age) were characterized by the frequent presence of distinct collagenous cellular nodules (cells: alpha-smooth-muscle actin+ and fibronectin+). Old HS (> or =3 years of age) showed widespread collagenization phenomena. The histological profile of K was not related to the age of the lesion and was characterized by the almost constant presence of abnormally thick, hyalinized collagen fibers, the presence of collagenous cellular nodules, and variable--albeit lower than in HS-- expression of alpha-smooth-muscle actin and fibronectin. Ultrastructurally, myofibroblasts were the predominant cell type in young and fully developed HS and in K. The immune-cell infiltrate was composed of CD3+, CD45RO+, CD4+, human lymphocyte antigen (HLA)-DR+, and lymphocyte function associated antigen (LFA)-1+ T lymphocytes, strictly associated with CD1a+/ CD36+, HLA-DR+, and intercellular adhesion molecule (ICAM)-1+ dendritic cells, both in HS and K. However, different amounts of immune cells were observed in relation to the type and age of the lesion, and these findings support the hypothesis that cell-mediated, major histocompatibility complex (MHC)-class II-restricted immune responses play an important role in the development of HS and K. PMID- 11407474 TI - Beta- and gamma-catenin expression in endometrial carcinoma. Relationship with clinicopathological features and microsatellite instability. AB - The activation of the adenomatous polyposis coli (APC)/beta-catenin/T-cell factor (Tcf) pathway due to beta-catenin gene mutation has been recently implicated in the development of some endometrial carcinomas. beta- and gamma-catenin are structurally and functionally related molecules that participate in cell adhesion and signal transduction. Nuclear accumulation of beta- and gamma-catenin have been related to the activation of the APC/beta-catenin/Tcf pathway. In this study, we investigate the immunohistochemical expression pattern (nuclear vs membranous) of beta- and gamma-catenin in 40 endometrial carcinomas and their correlation with clinicopathological features and microsatellite instability (MI) status. MI was detected at three or more loci in 12 tumors: 11 were endometrioid and one was non-endometrioid. Nuclear catenin expression was found in 13 carcinomas: ten carcinomas had nuclear beta-catenin expression and three carcinomas had nuclear gamma-catenin expression. The nuclear catenin expression pattern significantly correlated with the histological type, International Federation of Gynecology and Obstetrics (FIGO) grade, and the presence of a second neoplasm. Nuclear catenin expression was always observed in low-grade endometrioid carcinomas; it was also more frequently associated with a second carcinoma. No correlation was observed between the catenin expression pattern and the level of myometrial infiltration, stage, associated endometrial hyperplasia, the existence of a source of estrogenic stimulation, and MI. However, four of 13 endometrioid carcinomas in this series had both catenin nuclear expression and MI. These data suggest that at least two different neoplastic pathways can lead to endometrial carcinomas with an endometrioid phenotype. In one, MI would be a key event, while in the other, the APC/beta-catenin/Tcf signaling pathways could be activated. Probably, in some cases, both pathways could simultaneously occur. PMID- 11407475 TI - Endometrial carcinoma: association of steroid hormone receptor expression with low angiogenesis and bcl-2 expression. AB - In endometrial tissues, malignant change may be accompanied by a loss of hormone dependence which is, usually, reflected in a parallel loss of oestrogen and progesterone receptors (ER and PR). In this study, the steroid receptor status of 164 endometrial carcinomas was related to intratumoural angiogenesis and the apoptotic proteins bcl-2 and p53. Relationships to conventional histopathological features and patient survival were also sought. Immunohistochemistry was performed on formalin-fixed, paraffin-embedded tissues. The mean follow-up was 55 months (range 19-167 months). Specific nuclear staining for ER and PR was detected in 35% and 32% of endometrial carcinomas, respectively, and was very commonly co-expressed (P<0.0001). The failure of demonstrating a steroid receptor complement in endometrial neoplasms was, in general terms, an adverse prognostic sign. Thus, ER or PR loss was significantly associated with non-endometrioid carcinomas (ER P=0.01; PR P=0.004) and with deep myometrial invasion (ER P<0.0002), high intratumoural angiogenesis (PR P<0.01) and the absence of bcl-2 expression (PR P<0.005). There was a trend for patients with ER or simultaneous ER/PR expression to have an improved survival, but this association did not reach the level of statistical significance. In multivariate analysis (all stages), tumour cell type (endometrioid versus non-endometrioid carcinomas) and stage of disease were the only variables associated with prognosis (P=0.01 and P<0.0001, respectively), with tumour cell type retaining its independent prognostic value and within stage-I endometrial carcinomas (P=0.02). It is suggested that the loss of steroid hormone receptors in endometrial carcinomas is associated with a more aggressive phenotype and the switching-on of angiogenic pathways. PMID- 11407476 TI - Improvement of breast cancer prognostication using cell kinetic-based silver stainable nucleolar organizer region quantification of the MIB-1 positive tumor cell compartment. AB - Recently, it was stated that the proliferative activity (P) of a cell population could be indirectly calculated by multiplying the MIB-1 immunopositivity and silver-stainable nucleolar organizer region (AgNOR) features extracted exclusively in MIB-1 positive (pos.) nuclei: P=MIB-1 x AgNOR(MIB-1pos.). To study the prognostic significance of this hypothesis, MIB- immunohistochemistry and AgNOR staining were applied on a series of 89 cases of breast cancer with an 8 year follow-up period. The mean MIB-1 immunopositivity (MIB-1mean) was evaluated immunohistometrically on paraffin sections using a TV image analysis system CM-2 (Hund, Wetzlar, Germany). Later, a combined MIB-1/AgNOR staining was applied and evaluated using a TV image analysis system AMBA (IBSB, Berlin, Germany). The AgNOR features of 150 randomly chosen tumor nuclei were investigated, irrespective of their MIB-1 status (AgNOR count, AgNOR area). Later, a second measurement was performed on 100 MIB-1 positive tumor nuclei exclusively (AgNOR countMIB-1pos., AgNOR areaMIB-1pos.). AgNOR count and AgNOR countMIB-1pos. showed a different data distribution [2.7+/-0.7 (mean+/-SD) vs 3.9+/-1.1; r=0.315, P=0.014]. Similar results were obtained for AgNOR area and AgNOR areaMIB-1pos. (5.1+/-2.1 microm2 vs 7.5+/-2.4 microm2; r=0.501, P<0.001). Kaplan-Meier survival curves revealed significant differences for MIB-1mean (P=0.0018) and AgNOR areaMIB-1pos. (P=0.0340). In Cox models, both parameters provided independent prognostic information. Using their combination, the P, three groups of patients with statistically different survival could be separated (P=0.0014). Thus, the combination of MIB-1-immunopositivity and AgNOR measurements in MIB-1 positive nuclei appears to be more useful in breast cancer prognosis than the exclusive application of one of the two methods. By this combined application, probably effects of tumor biology are represented more precisely. PMID- 11407477 TI - Expression of the endothelin-B receptor in pigment cell lesions of the skin. Evidence for its role as tumor progression marker in malignant melanoma. AB - Endothelins (ETs) exert several functions in human melanocytes, including proliferation, dendrite formation, and melanin synthesis. Among the ET receptors, the non-selective endothelin-B (ETB) receptor is the major receptor in melanocytes and malignant melanoma (MM) cells. In spite of the important role of ETs and their receptors in the growth and differentiation of melanocytes, the distribution and expression levels of ETB receptors in tissue sections of benign and malignant pigment cell lesions is still unknown. We combined immunohistochemistry and reverse transcriptase-polymerase chain reaction (RT-PCR) to study ETB receptor expression in benign and malignant pigment cell lesions and in normal skin. Immunohistochemistry on paraffin-embedded tissue sections of 159 cases revealed a significant increase in intensity of ETB receptor expression from common nevi over dysplastic nevi and primary MM to metastatic MM. Quantitative PCR using realtime detection on 75 samples confirmed the immunohistochemical results. These data add the ETB receptor to the growing list of tumor progression markers in MM and suggest that ETs play a role in the progression of MM in the skin. PMID- 11407478 TI - Transforming growth factor beta production by spontaneous malignant mesothelioma cell lines derived from Fisher 344 rats. AB - We investigated whether transforming growth factor beta (TGF-beta) is involved in the growth of malignant mesothelioma (MM) cells in culture. TGF-beta production was examined in two mesothelioma cell lines (MeET-4 and -6) that were established from rat spontaneous MM in our laboratory. TGF-beta bioactivity in conditioned medium of these cell lines was analyzed using a CCL64 mink lung epithelial cell growth inhibition assay and found to be 30-70 times higher than that of normal rat mesothelial cells (MCs). The MM cell lines also showed considerably higher levels of TGF-beta mRNA expression when compared with MCs. The bioactivity and mRNA expression level were greater in MeET-4 than MeET-6. When MeET-4 was treated with antisense TGF-beta1 oligonucleotide (ODN), a significant decrease in both anchorage-dependent and -independent growth was observed. Treatment with exogenous TGF-beta resulted in no effects on the growth pattern of the MM cell lines, while proliferation of the MCs was slightly induced. It is considered that TGF-beta appears to be produced by rat spontaneous MM cells through an autocrine mechanism and could modulate the malignant growth of the tumor cells. PMID- 11407479 TI - Immunohistochemical patterns of human liver sinusoids under different conditions of pathologic perfusion. AB - This study reports the effects of altered hepatic perfusion on the sinusoidal bed and the phenotypic characteristics of sinusoidal endothelial cells (SECs). Sinusoids were studied by the application of endothelial cell markers (CD31, CD34, CD105, and ATZ 11) in lesions with localized increased perfusion (liver cell adenoma, focal nodular hyperplasia, and macroregenerative nodule), in chronic congestion, in decreased portovenous inflow (portal vein thrombosis), and in decreased arteriohepatic perfusion (obliterative arteriopathy in chronic allograft rejection). SECs react in a sensitive and uniform way to all investigated conditions of different pathologic liver perfusion: expression of CD31, CD34, and ATZ 11 by SEC is found in inflow areas, CD105-positive SECs are found at the end of the sinusoidal blood stream. This blood flow-orientated phenotypic shift of SECs was accompanied by a perisinusoidal accumulation of activated hepatic stellate cells and collagen IV. These findings are helpful in liver biopsies and provide new insights into the angioarchitecture of benign nodular lesions. PMID- 11407480 TI - Sebaceous gland metaplasia in intraductal papilloma of the breast. AB - We report here the first case of sebaceous gland metaplasia arising within an intraductal papilloma of the breast of a 70-year-old female. Several lobules and nests composed of clear cells closely resembling sebaceous glands of the skin were discovered within an intraductal papilloma of the breast. Squamous metaplasia was also noted in certain areas of the tumor. Immunohistochemically, the cells of the lobules and nests stained positively for monoclonal antibodies anti-cytokeratin 14 and epithelial membrane antigen. This study confirms a novel type of metaplasia of the breast. PMID- 11407481 TI - Acral myxoinflammatory fibroblastic sarcoma with unique clonal chromosomal changes. AB - Acral myxoinflammatory fibroblastic sarcoma is a rare tumor of the distal extremities. We present the hitherto unreported karyotypic abnormalities of this new entity. The tumor presented as a mass in the dorsum of the foot in a 53-year old woman and showed the typical virocyte-like and lipoblast-like cells in a myxoid and inflammatory background. Cytogenetic analysis revealed a complex karyotype with a reciprocal translocation t(1;10) (p22;q24) in addition to the loss of chromosomes 3 and 13. Fluorescence in situ hybridization with the 769E11YAC and BAC 31L5 and 2H23 probes showed the breakpoint to be located proximally to BCL10 and distally to GOT1 genes on chromosomes 1p22 and 10q24, respectively. The presence of these clonal chromosomal changes supports the neoplastic nature of acral myxoinflammatory fibroblastic sarcoma and underscores that it represents a separate entity. PMID- 11407482 TI - Gynecomastia in type-1 neurofibromatosis with features of pseudoangiomatous stromal hyperplasia with giant cells. Report of two cases. AB - We describe the histological finding in two cases of gynecomastia in patients with von Recklinghausen's disease. The histological and immunohistochemical features of the two cases were reviewed and compared with those of five cases of gynecomastia in men without clinical evidence of neurofibromatosis. In both patients bearing von Recklinghausen's disease, the breast stroma showed features consistent with pseudoangiomatous stromal hyperplasia (PASH). It was characterised by anastomosing empty spaces lined by spindle and multinucleated giant cells which were positive with CD34 and anti-vimentin antisera and negative with anti-FVIII and CD31 antisera. In two of five of the control cases without neurofibromatosis, the mammary stroma showed focal areas with features of PASH, but no multinucleated giant cells were present in any case. PASH with giant cells should be recognised as a feature of gynecomastia in von Recklinghausen's disease. The presence of multinucleated giant cells is very unusual and, although more cases have to be studied, these cells seem to be a feature of PASH occurring in patients with von Recklinghausen's disease. PMID- 11407483 TI - Metastasis from apocrine carcinoma of the breast to an endometrial polyp. PMID- 11407484 TI - An osteoclast-like giant cell tumor pattern in a mucinous cystadenocarcinoma of the pancreas with lymph node metastasis in a patient surviving over 10 years. PMID- 11407485 TI - Reminiscence and its relationship to attachment and personality in geropsychiatric patients. AB - The relationships between reminiscence functions and attachment styles; reminiscence and personality factors; and attachment and personality were examined in forty patients attending a geropsychiatric outpatient clinic. They were administered the Reminiscence Functions Scale, NEO-FFI, and the Relationship Questionnaire. Compared with insecurely attached older patients, securely attached older patients score higher on the teach/inform reminiscence function. Consistent with prior research, relationships were found between the extraversion personality factor and conversation reminiscence; and between the openness personality factor and both identity and problem-solving reminiscence functions. PMID- 11407486 TI - Age-related patterns in social networks among European Americans and African Americans: implications for socioemotional selectivity across the life span. AB - Socioemotional selectivity theory contends that as people become increasingly aware of limitations on future time, they are increasingly motivated to be more selective in their choice of social partners, favoring emotionally meaningful relationships over peripheral ones. The theory hypothesizes that because age is negatively associated with time left in life, the social networks of older people contain fewer peripheral social partners than those of their younger counterparts. This study tested the hypothesis among African Americans and European Americans, two ethnic groups whose social structural resources differ. Findings confirm the hypothesis. Across a wide age range (18 to 94 years old) and among both ethnic groups, older people report as many emotionally close social partners but fewer peripheral social partners in their networks as compared to their younger counterparts. Moreover, a greater percentage of very close social partners in social networks is related to lower levels of happiness among the young age group, but not among the older age groups. Implications of findings for adaptive social functioning across the life span are discussed. PMID- 11407487 TI - Networks in later life: an examination of race differences in social support networks. AB - Although there has been considerable interest in the effects of social support networks on various health outcomes for older adults, there has been little research directed toward the predictors of networks. In this study, we examine race differences in the determinants of social support network characteristics (size, frequency of interaction with network members, proportion of kin, and amount of support received and given to network members) using data from an older community sample drawn from the North Carolina site of the Established Populations for Epidemiologic Studies of the Elderly (EPESE) focusing on adults sixty-five and older (n = 4124). This research focuses on the extent to which race differences in network dimensions are present and whether these variations can be attributed to varying social structural positions held by African Americans and Whites. The results indicate that several race differences persist even when controlling for social structural variables. The structural argument and future implications are discussed. PMID- 11407488 TI - Social support and depression among elderly Chinese people in Hong Kong. AB - This study examines the association between social support and depressive symptomatology in a representative community sample of 1106 Chinese people in Hong Kong aged sixty years or older. Significant bivariate relationships were found between depression and all dimensions of social support including social network size, network composition, social contact frequency, satisfaction of social support, instrumental/emotional support, and helping others. Using multiple regression models, the authors found that at least one measure of these six dimensions of social support was associated with depressive symptomatology, even after controlling for socio-demographic, and functional disability. We found that social support from family is important for elderly Chinese people in Hong Kong, and satisfaction with support is a more important predictor of depression levels than other objective measures of network relationships. Lastly, it was found that material aid and instrumental support is more important in preventing depression for elderly individuals in Hong Kong than emotional support. PMID- 11407489 TI - Elderly individuals' reminiscences about the life-span development of their family. AB - Elderly individuals were asked to compose "family autobiographies" describing past events representing the beginning of their family, their family when the parents were middle-aged, and their family as it is today. Content analyses of the participants' recollections showed elderly adults' memories of the beginnings of their families were focused on episodes that illustrated the initial occurrence of family themes and values. In contrast, elderly individuals' recollections of their families when they were middle-aged and their families of today showed a tendency to attempt to identify continuing themes and trends from the episodes of their families' lives. These findings were taken to indicate individuals' reminiscences of their family lives changes as the relative importance of the families' developmental tasks changes. This pattern of findings was discussed in terms of Duvall's (1988) model of family developmental tasks. PMID- 11407490 TI - Exocellular enzyme activity of dermatophytes and other fungi isolated from ruminants in Southern Iraq. AB - Sixteen fungal species were isolated from 182 specimens collected from four ruminants (buffalo, camel, cattle and sheep) in Southern Iraq. Fungi represented by five species of dermatophytes and eleven species of other fungi were screened for the activity of four enzymes: keratinase. proteinase, lipase and amylase. Keratinase was found to be produced by all of the dermatophytes and non dermatophytes, except for Paecillomyces variottii and Scytalidium lignicola. However, high keratinase activity was expressed by the dermatophytic species particularly by Trichophyton mentagrophytes var. erinacei and Microsporum gypseum. Three dermatophytes viz. M. gypseum, T verrucosum and T. mentagrophytes var. nodulare were capable of producing protease, lipase and amylase. Although, T. mentagrophytes var. erinacei showed high protease activity, it did not produce lipase and amylase. On the contrary most of the non-dermatophytic species revealed protease and lipase activities higher than the dermatophytes. The Curvularia spp. isolates showed the highest protease and amylase activity, while Aspergillus parasiticus revealed the highest activity of lipase andamylase. No correlation was observed between enzyme activity and the growth rate of the examined fungi. PMID- 11407491 TI - In vitro evaluation of trichoderma and gliocladium antagonism against the symbiotic fungus of the leaf-cutting ant Atta cephalotes. AB - The antagonistic activity of Trichoderma and Gliocladium isolates against Attamyces sp., a symbiotic fungus of the leaf-cutting ant Atta cephalotes, was investigated. A. cephalotes cultures this fungus as the primary food source. Most of the Trichodema and Gliocladium isolates tested in vitro (82.6%) inhibited the Attamyces sp. mycelial growth. which was probably due to their colonization ability and competition for nutrients, both of them known mechanisms of some species of these genera. T. lignorum strain T-26 was the strongest inhibitor achieving a colonization of 23%. Microscopical observations indicate that the inhibitory effect was caused by an interaction that took place in close contact with the host hypha, causing wall deformation that led to the collapse of the turgor pressure. PMID- 11407492 TI - Alternaria brasiliensis sp. nov., a leaf pathogen on Phaseolus vulgaris. AB - Alternaria brasiliensis sp. nov., was isolated from leaves of common bean, Phaseolus vulgaris L., showing punctiform non concentric leaf spots of brown color. Besides the symptomatological differences, the new Alternaria species presents distinct type of arrangement of conidia chain, body and beak size. The disease was observed in Montanha county, State of Espirito Santo, Brazil, but not yet reported in the literature. The causal agent of the disease is now described by the first time. The epithet used here is referring to the country where the species has been found. PMID- 11407493 TI - Deterioration and spoilage of peanuts and desiccated coconuts from two sub Saharan tropical East African countries due to the associated mycobiota and their degradative enzymes. AB - A broad variety of fungi (84 species belonging to 36 genera) were identified with more taxa infesting peanut seed samples from two tropical countries (29 genera and 61 species) compared to those found in desiccated coconuts (20 genera and 55 species) on both DRBC and DG18 media. This may be due to the higher moisture levels in peanuts (5.07-7.97%) compared with coconuts (1.5-4.17%). More taxa and propagules were recovered on DG18 in both cases. The dominant fungi from both substrates on both isolation media were Aspergillus and Penicillium, with other fungi from only one substrate/medium. The aflatoxigenic species (A. flavus) dominated Kenyan samples more so than Ugandan samples on both substrates. However only 71.5% and 87.5% of the peanut kernels, on DRBC and DG18, respectively, were found to be infested with fungi. The aflatoxigenic species (A. flavus/parasiticus) were found in 75% of the samples, however only 15.75% and 13% of the kernels analyzed were infested. The most frequently isolated species from peanuts were A. niger followed by A. flavus and M. phaseolina. E. repens, E. amstelodami, E. rubrum and E. chevalieri dominated peanut seeds on DG18, and R. stolonifer, A. parasiticus, F. solani, L. theobromae and P. chrysogenum on DRBC. The mean count of fungal propagules in coconut samples were approximately 0.7 x 10(3) and 0.8 x 10(3) on DRBC and DG18, respectively, with a high proportion of those propagules recorded for the aflatoxigenic species (about 0. 17 x 10(3) and 0.25 x 10(3) colonies/g). The mycobiota of desiccated coconut was dominated by A. niger, A. flavus and P. chrysogenum. Also A. ochraceus, P. waksmanii, Paecilomyces variotii, P. islandicum and R. mucilaginosa were more frequent on DRBC, while, species of Cladosporium. Chrysosporium and Eurotium were more frequent on DG18. Enzyme indices (or the activities) for each specific strain, when determined after 5 and 8 days of incubation, proved to be similar. A recommendation is given. The proteolytic and lipolytic potentialities of the most commonly encountered species from both peanuts and coconuts were studied. The most interesting observation is that most of the positive isolates, in both commodities had high enzymic activity compared to those reported earlier for isolates of the same species. Such capabilities suggest that these commodities are expected to deteriorate, since climatic conditions in tropical areas favour fungal proliferation. Emphasis on the proper harvesting, drying, handling, transportation and/or storage; and also education of the populace, especially those are dealing with these foods, should be taken into consideration by the relevant authorities. The contaminated foods constitute a health hazard for human consumption. PMID- 11407494 TI - Identification and survival of the causal organism of leaf smut disease of cowpea in Nigeria. AB - Protomycopsis phaseoli (Ramak and Subram) is the causal agent of the cowpea leaf smut disease in Nigeria and not Entyloma vignae as claimed by some authors. This pathogen formed dark ash-grey to sooty-black lesions of 3-10 mm in diameter, while young lesions had yellow haloes. P. phaseoli produced dark reddish-brown chlamydospores that are globose to oval measured 23.8 microm, thick-walled and rugose. The chlamydospores germinated and produced globose vesicles. The pathogen grew on potato dextrose agar only when the leaf tissue was dipped in acidified water (1% H2SO4). The organism was slowly growing at 24-28 degrees C with snow white colour. Chlamydospores of P. phaseoli in infected cowpea leaves survived longer when buried in the soil for five months than when they were left on the soil surface for the same period at temperatures (26-27 degrees C) and humidity (70-82%) prevailing in Ibadan. Destruction of leaf debris before crop emergence, long period of rotation and no tillage cropping are suggested to prevent the onset and spread of leaf smut disease of cowpea. PMID- 11407495 TI - Evaluation of fungal growth on cellulose-containing and inorganic ceiling tile. AB - Buildings with poor indoor air quality (IAQ) frequently have many areas with surface fungal contamination. Studies have demonstrated that certain fungal genera (e.g., Cladosporium, Penicillium, and Stachybotrys) are able to grow on building materials such as wallpaper, drywall, and ceiling tiles, particularly after water damage has occurred. Due to the increasing awareness of sick building syndrome (SBS), it has become essential to identify building materials that prevent the interior growth of fungi. The objective of this study was to identify building materials that would not support the growth of certain fungal genera, regardless of whether an external food source was made available. The growth of three fungal genera (Cladosporium, Penicillium, and Stachybotrys) was evaluated on cellulose-containing ceiling tile (CCT) and inorganic ceiling tile (ICT). Both types of ceiling tile were exposed to environmental conditions which can occur inside a building. Our results show that ICT did not support the growth of these three fungal genera while CCT did. Our data demonstrate that ICT could serve as an ideal replacement for CCT. PMID- 11407496 TI - Neutralization of Latrodectus mactans and L. hesperus venom by redback spider (L. hasseltii) antivenom. AB - OBJECTIVE: To test the effectiveness of L. hasseltii (redback spider) antivenom in neutralizing the lethal effects of L. hesperus and L. mactans (North American black widow) venoms. METHODS: LD50 values for the L. hesperus and L. mactans venom preparations were determined. A prospective, randomized, double-blind antivenom efficacy experiment was then performed for each venom using a mouse envenomation model. The following treatments were premixed and incubated at 25 degrees C for 1 hour prior to intraperitoneal injection: 1) saline control + protein control, 2) saline control + L. hasseltii antivenom, 3) L. hesperus or L. mactans venom + protein control, and 4) L. hesperus or L. mactans venom + L. hasseltii antivenom. The study endpoints were time elapsed until death and survival at 24 hours. RESULTS: The mouse LD50 values for L. hesperus and L. mactans venoms were 0.64 mg/kg and 0.26 mg/kg, respectively. In the efficacy trial, all mice in group 3 (L. hesperus or L. mactans venom and protein control) died. In both experiments, all mice in group 4 (L. hesperus or L. mactans venom + antivenom) survived (p < 0.0001). CONCLUSION: This is the first study to derive mouse LD50 values for L. hesperus and L. mactans venom obtained by electrical stimulation of live adult spiders. Redback spider antivenom is effective in neutralizing the lethal effects of L. hesperus and L. mactans venoms in a mouse envenomation model. While this study is limited by the optimized premixing of antigen with antibody, it generates the hypothesis that redback antivenom would be effective in the treatment of latrodectism in humans caused by the two clinically relevant species of North American widow spiders. PMID- 11407497 TI - The safety and efficacy of antivenin Latrodectus mactans. PMID- 11407498 TI - Phentolamine reduces myocardial injury and mortality in a rat model of phenylpropanolamine poisoning. AB - BACKGROUND: Phenylpropanolamine produces dose-related, life-threatening cardiovascular, and central nervous toxicity from alpha-adrenergic overstimulation. Although some recommend the alpha-adrenergic antagonist, phentolamine, as treatment for such toxicity, its therapeutic efficacy has not been previously studied. We sought to determine if pretreatment with phentolamine could reduce acute myocardial injury and mortality in rats administered an overdose of phenylpropanolamine. METHODS: In the mortality arm of the study, 28 unanesthetized, male Wistar rats (14 animals per group) were randomized to receive an intraperitoneal injection of phentolamine (3 mg/kg) or an equal volume of normal saline diluent (control group). Twenty-five minutes later, all rats received an intraperitoneal injection of phenylpropanolamine (150 mg/kg). Mortality was compared at 24 hours. In the myocardial injury arm of the study, 20 unanesthetized rats (10 per group) were randomized to receive an intraperitoneal injection of phentolamine (3 mg/kg) or normal saline (control group). Twenty-five minutes later, all rats received an intraperitoneal injection of phenylpropanolamine (75 mg/kg). Seventy-two hours after phenylpropanolamine administration, all surviving animals were sacrificed and transverse sections of their hearts were graded histologically for injury by a blinded cardiac pathologist. RESULTS: Twelve rats died within 6 hours of phenylpropanolamine administration. Mortality was significantly lower in the phentolamine-pretreated rats (2/14; 14%) as compared to the control group (10/14; 71%; p = 0.006). The degree of myocardial injury was significantly lower in the phentolamine pretreated rats (0) as compared to the control group (1.4 +/- 1.6; p = 0.012). CONCLUSION: In this rat model, phentolamine pretreatment prevented acute myocardial injury and significantly reduced lethality from an intraperitoneal phenylpropanolamine overdose. PMID- 11407499 TI - Cardiovascular effects of buccal exposure to dermal nicotine patches in the dog: a comparative evaluation. AB - OBJECTIVE: Safety concerns have been raised over the possible effects of inappropriate exposure to transdermal nicotine patches. This study was initiated to determine whether placement of these products into the mouth could affect cardiovascular function. METHODS: In a series of 10 anesthetized beagle dogs, Nicoderm, Habitrol, ProStep I (Intact), ProStep D (Damaged) transdermal nicotine products or the Skoal Bandit smokeless tobacco plug were placed in the buccal cavity for 5 minutes. Systemic arterial blood pressure and the electrocardiogram were monitored for up to 90 minutes after exposure with blood samples at intervals during the first 10 minutes for plasma nicotine concentration. RESULTS: The systolic and diastolic arterial blood pressures and heart rate increased within 2 minutes of buccal exposure to either the intact or the damaged ProStep nicotine product. Ventricular arrhythmias were observed in 6 of 10 dogs exposed to the intact patch and 7 of 10 dogs exposed to the damaged patch during the period of maximal cardiovascular response. Modest increases in systemic blood pressure and heart rate were seen with the Nicoderm and Habitrol products but not with the Skoal Bandit. The increases in systemic arterial pressure and heart rate occurring after exposure to ProStep were significantly more severe than those observed after Nicoderm and Habitrol. Mean peak nicotine levels of 9.8 microg/mL (ProStep 1), 5.4 microg/mL (ProStep D), 3.4 microg/mL (Habitrol), 2.5 microg/mL (Nicoderm), and 0.12 microg/mL (Skoal Bandit) were detected within 2 to 10 minutes after buccal placement of the product. CONCLUSIONS: Certain transdermal nicotine patches, when applied to a nondermal site such as the buccal cavity for a short period (5 minutes) can rapidly provoke significant cardiovascular alterations (hypertension, tachycardia, and ventricular arrhythmias). The magnitude of the cardiovascular responses occurring after buccal exposure to a product such as ProStep could pose a risk to susceptible individuals. PMID- 11407500 TI - Rhabdomyolysis in isoniazid poisoning. AB - BACKGROUND: Rhabdomyolysis is one of the reported complications of isoniazid poisoning, but relevant data are limited. METHODS: A retrospective study was conducted on isoniazid poisoning cases seen at the Philippine General Hospital over 5 years (1992-1997). Patients excluded from the study were those who coingested other substances, including hepatotoxic and nephrotoxic drugs, those with underlying medical illnesses, and those without creatine phosphokinase muscle fraction determinations. RESULTS: Out of the 270 cases of isoniazid poisoning, 52 patient records were reviewed. Common clinical manifestations were seizures (100%), depressed sensorium (53%), and vomiting (45%). Laboratory results showed leukocytosis (74.5%), metabolic acidosis (29%), and impaired liver function tests (21%). Creatine phosphokinase muscle fraction was elevated in 59.6% of cases, beginning at a dose of 2.4 g. Values peaked on days 5 and 6 and declined on days 7 and 8. Statistically significant correlations were observed for the elevation of creatine phosphokinase muscle fraction with the duration/amount of drug ingested and the frequency of seizure. No correlation was observed between the frequency of seizures and elevated creatine phosphokinase muscle fraction nor between the time delay in consultation and elevation of creatine phosphokinase muscle fraction. CONCLUSIONS: The incidence of rhabdomyolysis in isoniazid poisoning was 3/100 cases a year. The findings suggesting its direct toxic effect on the muscles may not be clinically relevant. PMID- 11407501 TI - Time required for blood lead levels to decline in nonchelated children. AB - OBJECTIVE: To determine the time for a decline in blood lead to less than 10 microg/dL in nonchelated children who are enrolled in case management. STUDY DESIGN: Retrospective analysis of venous blood lead data of lead-poisoned children followed in a case management program designed to decrease lead exposure. Children were excluded if their blood lead had not yet declined to less than 10 microg/dL, if they received chelation therapy, or if they had not received follow-up for more than 15 months. We calculated the time between peak elevation of lead and decline to less than 10 microg/dL. Data were categorized based on the child's peak blood lead and season in which their peak blood lead occurred. Data were analyzed using ANOVA and linear regression. Kaplan-Meier survival analysis was used to describe data in population form. RESULTS: 579 patients were included in the analysis. Blood leads of 25-29, 20-24, 15-19, and 10-14 microg/dL required 24.0, 20.9, 14.3, and 9.2 months, respectively, to decline to less than 10 microg/dL. For continuous data, a linear relationship was described by the following equation: Time (# of months required to achieve a blood lead less than 10 microg/dL) = 0.845 x peak lead; p < 0.0001. Kaplan-Meier curves complement the findings in a population-based fashion. CONCLUSIONS: The mean time for blood lead to decline was linearly related to the peak in blood lead. The time for 50% of the blood lead to decline to less than 10 microg/dL was not linear and varied with peak lead. PMID- 11407502 TI - Death following cupric sulfate emesis. AB - CASE HISTORY: A 25-year-old woman who had ingested about 20 tablets of diazepam 2.5 mg in a suicide attempt was given cupric sulfate 2.5 g in 1750 mL water as an emetic, but died 3 days later. On autopsy, death was attributed to acute hemolysis and acute renal failure due to copper poisoning. Copper concentrations were 5.31 microg/mL in whole blood, 19.0 microg/g in the liver, 8.9 microg/g in the kidney, 1.1 microg/L in the brain, 1.1 microg/g in the gastric wall, 1.5 microg/g in the jejunal wall, 0.3 microg/g in the colon wall, 4.6 microg/g in the gastric contents, and 12.6 microg/g in the intestinal contents (fresh weight). This case and 10 others from the Chinese medical literature provide additional evidence that cupric sulfate is a corrosive poison and contraindicated as an emetic. PMID- 11407503 TI - Phosphine exposure from a methamphetamine laboratory investigation. AB - BACKGROUND: Law enforcement personnel may be exposed to a variety of hazardous materials during investigation of clandestine methamphetamine laboratories. CASE REPORT: A 28-year-old forensic specialist investigating a methamphetamine lab was exposed to phosphine without respiratory protection at approximately 2.7 ppm for 20-30 minutes. Shortly following exposure she developed dizziness, cough, headache, and diarrhea, although initial medical evaluation within 1-2 hours was unremarkable. Pulmonary examination at 4 and 7 days postexposure revealed bilateral rhonchi. The cough was worse with exertion, and persisted despite beta agonist and steroid inhaler treatment. CONCLUSIONS: This is apparently the first published case of symptomatic occupational phosphine exposure in a law enforcement officer during investigation of a methamphetamine laboratory. If phosphine exposure is suspected, the possibility of delayed pulmonary toxicity should be recognized. PMID- 11407504 TI - Pediatric acetaminophen overdose. PMID- 11407505 TI - Acetaminophen toxicity nomogram error: Barkin's Textbook of Pediatric Emergency Medicine. PMID- 11407506 TI - Fulminant hepatic failure due to acetaminophen poisoning may be less common in Hong Kong. PMID- 11407507 TI - Pneumonitis following inhalation of a commercially available water repellent. PMID- 11407508 TI - False-positive amphetamine screen following a trazodone overdose. PMID- 11407509 TI - Duplicate submission and scientific fraud: the Annals of Surgical Oncology perspective. PMID- 11407510 TI - The future for staging of primary tumors. PMID- 11407511 TI - Treating colon cancer with a melanoma vaccine? Preposterous? PMID- 11407512 TI - CancerVax, an allogeneic tumor cell vaccine, induces specific humoral and cellular immune responses in advanced colon cancer. AB - BACKGROUND: The immunogenicity of the polyvalent tumor cell vaccine CancerVax has been correlated with the survival of patients receiving active immunotherapy for melanoma. Because the various antigens expressed on the vaccine are common to colon adenocarcinoma cells, we examined the survival impact of immune responses elicited by CancerVax in patients with advanced colon cancer refractory to standard therapy. METHODS: Twenty-seven patients with American Joint Committee on Cancer (AJCC) stage IV colorectal adenocarcinoma were entered prospectively into the study. CancerVax was coadministered with bacille Calmette-Guerin (BCG) for the first 2 weeks of vaccine treatment. Blood was drawn at the start of therapy and every 2 weeks thereafter to measure serum titers of immunoglobulin (Ig)G and IgM against TA90 (a 90-kD immunogen common to colon cancer and CancerVax cells) and against purified protein derivative (PPD), a nontumor control antigen. Cellular immune responses were evaluated by delayed-type hypersensitivity (DTH) reaction to vaccine cells and to PPD. Mean follow-up time was 17.5 months. RESULTS: There was a significant (P = .0001) increase in anti-TA90 IgG and IgM titers and in DTH response to vaccine cells. Humoral and skin responses to TA90 did not correlate with responses to PPD (P = .199 for IgM, P = .958 for IgG, and P = .149 for DTH). This suggests that these responses are not a manifestation of general immune competence. The median overall survival (OS) was 21.9 months for the entire group. Overall survival was higher among patients whose IgMTA90 titer was >800 (P = .003) or whose disease-free interval exceeded 12 months (P = .031). Multivariate Cox regression analysis-using age, sex, disease-free interval, disease status, extent of metastasis, humoral responses, and DTH responses-found only peak IgMTA90 titer to be a significant predictor of overall survival (P = .0365). CONCLUSIONS: CancerVax can induce measurable humoral and cellular immune responses to tumor-associated antigens in patients with advanced-stage colon cancer. These responses correlate with overall survival. This novel therapeutic regimen for patients with advanced colon cancer merits further investigation. PMID- 11407513 TI - Impact of splenectomy for lymph node dissection on long-term surgical outcome in gastric cancer. AB - BACKGROUND: In the treatment of gastric cancer, splenectomy is performed for effective lymph node dissection around the splenic artery and splenic hilum. The purpose of this study was to clarify the long-term outcome of splenectomy in the treatment of gastric cancer. METHODS: The effect of splenectomy on recurrence and prognosis was examined in a retrospective analysis of 665 patients who had undergone curative total gastrectomy for gastric carcinoma from 1987 to 1996. The risk factors associated with recurrence and prognosis were investigated by univariate and multivariate analysis. RESULTS: The splenectomy group showed more advanced lesions and a higher recurrence rate than the spleen-preserved group. However, after adjusting for the TNM (tumor, node, metastasis) stage, there was no significant difference in recurrence rate and pattern between the two groups. Logistic regression analysis revealed that gross type, serosal invasion, and nodal metastasis were independent risk factors for recurrence while splenectomy was not. When comparing patients with the same TNM (tumor, node, metastasis) stages, no significant difference in the 5-year survival rates was apparent. Multivariate analysis demonstrated that age, serosal invasion, and nodal metastasis were independent prognostic factors whereas splenectomy was not. CONCLUSIONS: These data suggest that splenectomy for lymph node dissection in gastric cancer is not effective regarding long-term patient prognosis. PMID- 11407514 TI - Clinical significance of K-Ras mutations in intraoperative tumor drainage blood from patients with colorectal carcinoma. AB - BACKGROUND: Recurrent and metastatic carcinoma of the colorectum remains a major problem. This may be ascribed to the presence of micrometastasis at diagnosis. The purpose of this study was to analyze prospectively the clinical value of detecting K-ras mutations in the perioperative circulating blood from patients with colorectal carcinoma. METHODS: Twenty-four patients whose tumor carried mutations in codon 12 of the K-ras gene were studied for the presence of cancer cells in perioperative blood samples, in particular, tumor drainage samples. A detection assay using CD45 immunomagnetic separation plus nested mutant allele specific amplification (MASA) was performed. RESULTS: K-ras mutations in CD45 negative cells in tumor drainage blood were detected in 7 (29.2%) of 24 patients. There was no significant relationship between the presence of a K-ras mutation and clinicopathological features. Four (57.1%) of the seven patients with a positive K-ras mutation in drainage blood had early recurrent disease. Of the 17 patients with no K-ras mutation, none developed metastatic disease. The recurrence rate of the K-ras mutation positive group was higher than that of the K-ras mutation negative group (P < .01). There was a significant difference, regarding prognosis, between K-ras mutation positive and negative groups (P < .01). CONCLUSIONS: This preliminary study demonstrates that the detection of circulating cancer cells in the tumor drainage blood by our new assay system may provide a predictor of recurrence and metastasis of colorectal cancer. PMID- 11407515 TI - Adenocarcinoma of the lower third of the rectum: metastases in lymph nodes smaller than 5 mm and occult micrometastases; preliminary results on early tumor recurrence. AB - BACKGROUND: The number of examined lymph nodes and metastases in lymph nodes smaller than 5 mm (small lymph nodes) are a determining factor in the stage of rectal cancer although the clinical significance of occult micrometastases is controversial. We are reporting our preliminary results on the identification and prognostic utility of metastases in small lymph nodes and occult micrometastases. METHODS: We searched small metastatic lymph nodes in 101 cases of adenocarcinoma of the lower third of the rectum. We used the manual technique to dissect mesorectal fat and occult micrometastases in the lymph nodes of 52 Dukes' A and B patients, using a pool of anticytokeratin antibodies. RESULTS: Forty-five percent of the metastatic lymph nodes were smaller than 5 mm in diameter and determined the Dukes' stage in 15 (30.6%) of 49 Dukes' C patients. Occult micrometastases were found in 21 (40.4%) patients: five recurred but vascular invasion, positive distal margin of the rectum, and positive circumferential margin of the mesorectum were present. CONCLUSIONS: Small metastatic lymph nodes, vascular invasion, positive distal margin of the rectum, and positive circumferential margin of the mesorectum were found to be more important than occult micrometastases in predicting early recurrence of rectal cancer. PMID- 11407516 TI - p53, BCL-2, and Ki-67 expression according to tumor response after concurrent chemoradiotherapy for advanced rectal cancer. AB - BACKGROUND: Concurrent chemoradiotherapy (CCRT) for locally advanced rectal cancer is an important modality for curative resection, but tumors show wide spectrum response. The purpose of this study was to investigate any correlation among related genetic mutations, proliferative index, and tumor response after CCRT. METHODS: This study included 23 patients with rectal cancer, who were preoperatively staged as at least T3 N1 or T4 (determined by transrectal ultrasonography and MRI). Enrolled patients were given 5-FU 450 mg/m2/day and leucovorin 20 mg/m2/day intravenously for 5 days during weeks 1 and 5 of radiotherapy (45-54 Gy). Surgical resection was performed 4 weeks after completion of the scheduled treatment. Tumor response was classified as CR (complete response), PR (partial response: 50% diminution of tumor volume and downstaging), and NR (no response). Paraffin-embedded tissue obtained before chemoradiotherapy was studied by immunohistochemical staining for p53, BCL-2, and Ki-67. The extent of tumor response was correlated with proliferative activity and was measured by immunostaining Ki-67 proliferative antigen and the expression of p53 and BCL-2 oncoproteins. RESULTS: All patients were resectable. CR was obtained in 4 patients, PR in 10 patients, and NR in 9 patients. The p53 mutation was noted in 16 patients: NR in 5 patients, PR in 9 patients, and CR in 2 patients (P = .638). BCL-2 expression was noted in 11 patients: NR in 4 patients, PR in 3 patients, and CR in 4 patients (P = .799). The Ki-67 labeling index was NR: 615.4+/-47.2; PR: 663.2+/-20.4; and CR: 765.5+/-58.3 (CR + PR vs. NR, P = .029). CONCLUSIONS: Immunohistochemical expression of p53 and BCL-2 does not correlate with tumor response after CCRT, but Ki-67 labeling may be a useful parameter for radiosensitive tumors selected for CCRT. PMID- 11407517 TI - Feasibility of breast preservation in the treatment of occult primary carcinoma presenting with axillary metastases. AB - BACKGROUND: The objective of the study was to compare the treatment outcomes in patients with occult primary carcinoma with axillary lymph node metastasis who were treated with mastectomy or with intent to preserve the breast. METHODS: From 1951 to 1998, 479 female patients were registered with axillary lymph node metastasis from an unknown primary. After clinical workup, including mammography, 45 patients retained this diagnosis and received treatment for T0 N1-2 M0 carcinoma of the breast. Clinical and pathological data were collected retrospectively, and survival was calculated from the date of initial diagnosis using the Kaplan-Meier method. Median follow-up time was 7 years. RESULTS: Median age was 54 years (range, 32-79). Clinical nodal status was N1 in 71% and N2 in 29% of the patients. Surgical treatment was mastectomy in 29% and an intent to preserve the breast in 71% of the patients. Locoregional radiotherapy was used in 71% and systemic chemoendocrine therapy was used in 73% of the patients. Of the 13 mastectomy patients, only one had a primary tumor discovered in the specimen. Two patients (4%) were ultimately diagnosed with lung cancer and neuroendocrine tumor. No significant difference was detected between mastectomy and breast preservation in locoregional recurrence (15% versus 13%), distant metastases (31% versus 22%), or 5-year survival (75% vs. 79%). Regardless of surgical therapy, the most important determinant of survival was the number of positive nodes. Five year overall survival was 87% with 1-3 positive nodes compared with 42% with > or =4 positive nodes (P < .0001). CONCLUSIONS: Occult primary carcinoma with axillary metastases can be treated with preservation of the breast without a negative impact on local control or survival. PMID- 11407518 TI - Factors correlating with lymph node metastases in patients with T1 breast cancer. AB - BACKGROUND: Identification of reliable predictors of axillary metastases (ALNM) may be useful in selecting appropriate management for patients with T1-size breast cancer. This study was undertaken to determine the degree of correlation between ALNM and several variables, including age, race, menopausal status, palpability, tumor size, positive margin on initial excision, histology, grade, lymphatic invasion (LI), estrogen receptor status (ER), progesterone receptor status, S-phase, and ploidy. METHODS: Data from 1416 patients with T1 breast cancers treated at Columbia-Presbyterian Medical Center between 1989 and 1998 was reviewed. Patients with multifocal tumors were excluded. RESULTS: Mean patient age was 57.5 years (SD = 12.0); 65% of the patients were postmenopausal. One hundred thirty-one patients with Tla (< or =0.5 cm), 435 with T1b (0.6-1.0 cm), and 850 patients with T1c (1.1-2.0 cm) lesions were studied. The overall rate of ALNM was 23%. AM was identified in 11% of T1a, 15% of T1b, and 29% of T1c patients. Statistically significant factors from univariate analysis were age, palpability, skin changes, tumor size, LI, histology, grade, ER status, and positive margin on initial excision. CONCLUSIONS: Axillary staging by either sentinel lymph node biopsy or level I/II axillary dissection is indicated for most T1 breast cancer patients. Omission of axillary staging can be considered for highly selected patients with T1a cancers. PMID- 11407519 TI - Sentinel lymph node detection for breast cancer: which patients are best suited for the patent blue dye only method of identification? AB - BACKGROUND: The objectives of this study were, first, to define the preoperative criteria for using solely the blue dye method and, second, to decrease its operator dependence in predicting axillary lymph node status. METHODS: Two hundred fifty-three women consecutively identified with operable breast cancer underwent sentinel lymph node (SLN) detection by the patent blue dye method followed by completion axillary lymph node dissection. A standard pathological examination was performed for all SLN. Then, a pathological color quality assessment (PCQA), which checked for the presence of the blue dye, was performed on the paraffin blocks of the nonmetastatic SLN. Six preoperative identifiable variables likely to influence the detection rate were examined. RESULTS: The surgical detection (sd) rate was 84% (213 of 253) and the PCQA rate was 73% (185 of 253). Only breast size (sd, P = .0005; PCQA, P = .0007) and body mass index < or =30 (sd, P = .005; PCQA, P = .0007) were significant for SLN identification. Multivariate analysis revealed two independent factors influencing SLN identification: breast size (sd, P = .0001; PCQA, P = .002) and the timing of injection-injection prior to lumpectomy (sd, P = .04). CONCLUSIONS: The optimal patient features for identifying the SLN by the patent blue dye method are small or medium-sized breasts, low body fat, and that the procedure is carried out prior to tumor excision. The PCQA offers a useful second assessment of the surgically removed SLN, introducing an independent element of quality control. PMID- 11407520 TI - Micrometastasis to in-transit lymph nodes from extremity and truncal malignant melanoma. AB - BACKGROUND: The sentinel lymph node (SLN) is the first lymph node in the regional nodal basin to receive metastatic cells. In-transit nodes are found between the primary melanoma site and regional nodal basins. To date, this is one of the first reports on micrometastasis to in-transit nodes. METHODS: Retrospective database and medical records were reviewed from October 21, 1993, to November 19. 1999. At the UCSF Melanoma Center, patients with tumor thickness > 1 mm or < 1 mm with high-risk features are managed with preoperative lymphoscintigraphy, selective SLN dissection, and wide local excision. RESULTS: Thirty (5%) out of 557 extremity and truncal melanoma patients had in-transit SLNs. Three patients had positive in-transit SLNs and negative SLNs in the regional nodal basin. Two patients had positive in-transit and regional SLNs. Three patients had negative in-transit SLNs but positive regional SLNs. The remaining 22 patients were negative for in-transit and regional SLNs. CONCLUSIONS: In-transit SLNs may harbor micrometastasis. About 10% of the time, micrometastasis may involve the in transit and not the regional SLN. Therefore, both in-transit and regional SLNs should be harvested. PMID- 11407521 TI - Effect of sodium thiosulfate on cisplatin removal with complete hepatic venous isolation and extracorporeal charcoal hemoperfusion: a pharmacokinetic evaluation. AB - BACKGROUND: Complete hepatic venous isolation and extracorporeal charcoal hemoperfusion (HVI.CHP) can limit systemic exposure to high-dose chemotherapeutic agents when given by hepatic arterial infusion (HAI). The purpose of this study was to determine if the concomitant use of sodium thiosulfate (STS) could further expand the advantages of pharmacologic delivery of HVI.CHP for cisplatin (CDDP) during HAI chemotherapy. METHODS: CDDP (4mg/kg) was administered over 20 minutes via HAI under conditions of HVI.CHP in 14 mongrel dogs. HVI.CHP was performed for 30 minutes after initiation of HAI. During CDDP infusion, 7 dogs each received 400 mg/kg STS (a 100-fold molar ratio to CDDP) over 20 minutes via the prefilter (STS group) circuit line, while the remaining 7 dogs (controls) received no STS. Blood samples were taken serially from the prefilter circuit line (hepatic venous blood), postfilter line, and the left carotid artery (systemic blood). The free and total CDDP concentrations in these samples were determined by flameless atomic absorption spectrophotometry. RESULTS: During 20 minutes HAI of CDDP, the mean CDDP extraction ratios (ER) by CHP filter were always higher in the STS group than in the control group, regardless of the form (free or total) of CDDP. The differences between the STS and control groups in the extraction ratios of free and total CDDP were significant at all time points measured (P < .05). Consequently, systemic exposure to CDDP, as assessed by area under the time concentration curve of total CDDP, was significantly lower in the STS group than in the control group (P < .05). CONCLUSIONS: These results indicated that concomitant STS infusion could further increase the effect of HVI.CHP on CDDP removal after HAI. PMID- 11407522 TI - Association of enhanced cyclooxygenase-2 expression with possible local immunosuppression in human colorectal carcinomas. AB - BACKGROUND: Prostaglandin (PG) E2 has an influence on antitumor lymphocyte reactions and causes local immunosuppression at tumor sites. The contribution of cyclooxygenase (COX), a key enzyme in PGE2 synthesis, to this effect is still unclear. We examined if cyclooxygenase (COX)-2 is involved in local immunosuppression in human colon carcinoma cell lines and in clinical tumor specimens. METHODS: PGE2 concentrations were measured in culture media from a highly COX-2-expressing human colon carcinoma cell line (CE-1) and other cell lines. Lymphocyte proliferation in response to a mitogen was used to evaluate immunosuppression in tumor cell-lymphocyte cocultures with and without selective COX-2 inhibitor NS-398. We also evaluated expression of COX-2 mRNA in surgical specimens of colorectal carcinoma by reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein by immunohistochemistry, correlating COX-2 expression with clinicopathologic features. RESULTS: CE-1 cells produced large amounts of PGE2, which was significantly inhibited by NS-398. The proliferation index of lymphocytes cocultured with CE-1 cells was significantly less than that of control lymphocytes; again, this effect was inhibited by NS-398. While human colorectal carcinoma tissue expressed more COX-2 mRNA and protein than nonneoplastic tissue, no significant correlation was found between COX-2 levels and clinicopathologic features. CONCLUSIONS: Overexpression of COX-2 in colon cancer may cause local immunosuppression, and COX-2 inhibitors might be therapeutically useful against these tumors. PMID- 11407523 TI - Ultrastructural morphometry of nucleoli: potential usefulness for objective grading of clear cell renal cell carcinoma. AB - The authors attempted to determine the potential prognostic value of several ultrastructural morphometric parameters, including nuclear, nucleolar, and cytoplasmic features, that could be used in the objective and reproducible histological grading of clear cell renal cell carcinoma. Several nuclear and cytoplasmic parameters were assessed by ultrastructural morphometry in 26 consecutive cases of clear cell renal cell carcinoma. The nuclear and nucleolar sizes, the number of nucleoli per nuclear section and the number of marginated nucleoli, Fuhrman's nuclear grade, and Robson's stage were recorded. In addition, the proportion of cytoplasmic components was semiquantitatively estimated and compared to light microscopic appearance. Follow-up ranged from 5 to 15 years (mean = 10 years). Statistical evaluations were performed by means of the Pearson or Spearman correlation coefficient tests, and differences in survival were estimated, using the Mantel-Cox proportional risk method. Differences in survival among patients with a mean nuclear area over and under 160 microm2, and among those with a mean nucleolar area over and under 10 microm2, were statistically significant. (Cutoff points were selected at the median value for both parameters; Mantel-Cox test: chi2 = 7.102, p < .01; and chi2 = 11.096, p < .001, respectively). Fuhrman's nuclear grade (p < .01) and tumor stage at diagnosis (p < .001) were also related to survival. These data suggest that, out of all the ultrastructural morphometric features, nucleolar area is the most useful in the reproducible and accurate grading of clear cell renal cell carcinoma. PMID- 11407524 TI - Electron microscopic detection of copper in the liver of two patients with morbus Wilson by EELS and EDX. AB - A 20-year-old male patient with morbus Wilson was liver transplanted because of terminal failure of liver function. The explanted liver showed a strong macronodular cirrhosis as typically seen in Wilson disease. There were visible granular accumulations in the hepatocytes after the rubeanic acid or rhodanine method for histochemical detection of copper. The electron microscopic studies on ultrathin sections revealed numerous electron-dense lysosomes and residual bodies. The elemental analysis in transmission electron microscope (TEM) with electron energy loss spectroscopy (EELS) and in scanning electron microscope (SEM) with energy dispersive x-ray analysis (EDX) showed copper-specific signals of electron-dense accumulations inside these dark lysosomes and residual bodies. In a second case, Wilson disease was diagnosed after autopsy of a 31-year-old patient by liver electron microscopy and EELS; strong electron-dense lysosomes and residual bodies with positive copper signals were found inside hepatocytes. For negative control, hepatocytes with iron accumulation after idiopathic hemochromatosis and liver cirrhosis were also analyzed by EELS in TEM, which showed strong iron, but only a few or no copper signals. Atomic absorption spectroscopy (AAS) in 16 liver samples of healthy and cirrhotic liver revealed only in both cases of Wilson disease a strong increased copper concentration higher than 100 microg Cu/g. The electron microscopic detection of copper containing hepatocytic lysosomes is helpful for the diagnosis of early stages of Wilson disease in addition to the quantification of hepatic copper by AAS. PMID- 11407525 TI - Ultrastructural of the bronchial epithelium in chronic inflammation. AB - Light-optic and electron microscopic study of the bronchial biopsy specimens from 208 patients with primary and secondary chronic bronchitis was carried out. Variants of the bronchial epithelium states were described and classified according to characteristics of differentiation processes. As a result, groups with relatively normal structure, with deviated differentiation processes (hyperplasia of goblet glandular cells, hyperplasia of basal epithelial cells, hypoplasia and degeneration of goblet glandular cells), with incomplete differentiation processes (proliferative transformation and transitory state), and with altered differentiation processes (squamous metaplasia and atrophy) were described. These variants of the bronchial epithelium states are not only indicative of different levels of change or structural disorders, but may also represent a dynamic scheme of its consecutive changes during the development of chronic inflammatory process. PMID- 11407526 TI - Unidentified bacterial microorganisms entrapped within blood capillary spaces of tissue from different epidemiological types of Kaposi's sarcoma. AB - Tissue specimens of different epidemiological types of Kaposis sarcoma (KS) from various geographical regions were investigated by transmission electron microscopy. Freshly fixed KS biopsies originated from 9 German patients: 3 classic KS cases, 5 AIDS-associated KS cases, and 1 atypical classic KS case. Additionally, KS autopsy material from the brain of a German AIDS patient was examined. Further biopsies came from 29 Ugandan patients: 16 endemic KS cases and 13 AIDS-associated KS cases. While investigating the ultrastructure, we discovered relatively small-sized bacterial microorganisms within blood capillary spaces of tumor tissue from 5 KS cases of different epidemiological type. The microorganisms often occurred in clusters. They were of coccoid-bacillary form and limited by a wrinkled multilayered cell wall. Many of them were encapsulated. They were not observed outside of the capillary lumen. The bacterial structures were often seen attached to capillary endothelial cells, which sometimes showed blistering into the capillary lumen. The observed bacterial microorganisms obviously represented agents of a bloodstream infection and must have been entrapped and accumulated within capillary spaces of KS tissue. The bacteria, which had an almost identical morphology in all 5 KS cases, could not be identified. If they are of pathogenic significance, it remains unknown. PMID- 11407527 TI - Duodenal periampullary gangliocytic paraganglioma: report of two cases with immunohistochemical and ultrastructural study. AB - We report two cases of Gangliocytic Paraganglioma (GP) of the ampulla of Vater occurring in a 63-year-old and a 34-year-old individual. The patients were both admitted for a long history of intermittent gastrointestinal bleeding and abdominal discomfort, with no other symptoms. At endoscopy, the GP appeared as a polypoid, ulcerated mass in the ampullar region, measuring 2.5x1.8 and 2 cm, respectively. Microscopically, the tumors showed similar features and were composed of epithelial cells (more than 50%), spindle cells, and ganglion-like cells. The epithelial cells showed clear cytoplasm and formed nests (zellballen or paraganglioma-like groups), and less frequently, cords (carcinoid-like), extending to mucosa and submucosa. Ganglion cells were sparse, constantly associated with the spindle cells. Both epithelial and ganglion cells were synaptophysin, chromogranin A, and anti-neurofilament immunoreactive. The spindle cells were all S-100 positive. Ultrastructural studies revealed dark and light cells, rare elongated cellular processes, secretory granules, and fine fibrils resembling neurofilaments. The histogenesis of GP is still a matter of debate, however its neoplastic nature is supported by the occasionally reported malignant evolution. PMID- 11407528 TI - Solid papillary carcinoma of the breast: report of two cases. AB - Two cases of solid papillary carcinoma of the breast were pathologically studied. Subjects included two female patients: a 76-year-old (Case 1) and a 43-year-old (Case 2). Both cases showed solid and papillary proliferation of spindle cells in expanded ducts, perivascular pseudorosettes, eosinophilic fine granules in an abundant cytoplasm, intracellular mucin production, and positivity for chromogranin A. Case 1 showed an aggregation form involving multiple ducts, admixture of signet-ring like cells, mucin pools, and small and large electron dense granules and flocculent granules in electron microscopy. Case 2 showed a solitary and compact form in a dilated duct and an interlacing bundle pattern. PMID- 11407529 TI - Renal oncocytoma with intracytoplasmic lumina: a case report with ultrastructural findings of "oncoblasts". AB - The authors present a case of renal oncocytoma with numerous intracytoplasmic lumina in a 45-year-old woman, with an emphasis on the ultrastructural findings of so-called oncoblasts. The tumor was located in the upper pole of the left kidney, measuring about 3.3 cm in diameter. Histologically, it was composed of solid nests or acinar growths of so-called oncocytic tumor cells with numerous intracytoplasmic lumina and scattered foci of so-called oncoblasts. The luminal surface was positive for Hale's colloidal iron stain. A very small number of glycogen-containing cells were found scattered in a few nests. Immunohistochemically, tumor cells were positive for cytokeratin and epithelial membrane antigen, but negative for vimentin and carcinoembryonic antigen. The Ki 67 labeling index was 1.2%. All of the Ki-67-positive cells were oncocytes. So called oncoblasts were negative for Ki-67. Ultrastructural examination revealed that the predominant tumor cells had cytoplasm packed with round mitochondria and the mitochodria had lamellar long cristae. So-called oncoblasts showed scant cytoplasm with a moderate number of small mitochondria. Some of them showed pyknosis which can be regarded as mitochondrial involution. The authors believe that so-called oncoblasts are damaged or involuted oncocytes rather than precursors of oncocytes. PMID- 11407530 TI - Ultrastructural evidence of in vivo phagocytosis of Helicobacter pylori. PMID- 11407531 TI - Case for the panel. Smooth tubular aggregates, presumably Golgi in nature, in a gastric carcinoma. PMID- 11407532 TI - Ultrastructural features of apoptosis in human pituitary adenomas. AB - Although several recent studies deal with various molecular aspects of apoptosis, or programmed cell death, very little information is available on the ultrastructural changes associated with apoptosis in the adenohypophysis and its role in the regulation of pituitary adenoma growth and progression. This paper describes the distinct ultrastructural sequences that develop during the various phases of the apoptotic process. The study is based on the ultrastructural investigation of more than 8,000 surgically removed pituitary biopsies, which were examined by histology and immunocytochemistry for diagnostic purposes. No apoptosis was found in normal adenohypophysis and it is also a rare event in pituitary adenomas. When present, adenomatous adenohypophysial cells exhibit common and characteristic apoptotic changes. The ultrastructural alterations of membraneous organelles associated with apoptosis are similar to those previously reported in other tissues. It is noteworthy that apoptosis is clearly distinguishable from the ubiquitous dark cells denoting the common way of cell death. The findings suggest that apoptosis in pituitary adenomas is not a random event. Practically every specimen containing multiple apoptotic cells represents corticotroph adenoma. Occasional examples occur in lactotroph or gonadotroph adenomas. Although electron microscopic specimens are admittedly small, the large number of investigated cases gives credence to the observations. PMID- 11407533 TI - Chronic heat-induced skin lesions (erythema ab Igne): ultrastructural studies. AB - Erythema ab igne (EI) is an uncommon skin lesion caused by mild and repeated exposure to infrared sources. The aim of this study was to investigate the ultrastructural alterations in this condition. The ultrastructural study was carried out on 5-outpatients who presented typical EI of their exposed sites. Skin punch biopsies were processed for standard electron microscopy. The epidermis was hyperpigmented, with focal regressive changes of basal keratinocytes. An apparent functional activation of melanocytes with numerical increase of dendritic processes was also observed. The dermis showed abundant melanophages and occasional elastic fiber alterations similar to actinic elastosis. No alterations consistent with preneoplastic skin conditions were observed. The ultrastructural findings associated with EI seem to be nonspecific and consistent with moderate regressive changes of keratinocytes as well as a consensual melanocytic activation and elastic fiber modifications. Similar alterations can be observed in chronic actinic skin damage. This condition is presumably more benign than the ultraviolet exposure. The association of EI and premalignant skin lesions, though occasionally described, seems relatively infrequent. PMID- 11407534 TI - Uncombable hair (cheveux incoiffables, pili trianguli et canaliculi) syndrome: brief review and role of scanning electron microscopy in diagnosis. AB - Uncombable hair syndrome was first described some 3 decades ago as "cheveux incoiffables" and is also known as spun-glass hair and pili trianguli et canaliculi. Both inherited (autosomal dominant and recessive with variable levels of penetrance) and sporadic forms of uncombable hair syndrome have been described, both being characterized by scalp hair that is impossible to comb due to the haphazard arrangement of the hair bundles. A characteristic morphologic feature of hair in this syndrome is a triangular to reniform to heart shape on cross-sections, and a groove, canal or flattening along the entire length of the hair in at least 50% of hairs examined by scanning electron microscopy. Most individuals are affected early in childhood and the hair takes on a spun-glass appearance with the hair becoming dry, curly, glossy, lighter in color, and progressively uncombable. Only the scalp hair is affected. Several conditions are associated with uncombable hair, such as ectodermal dysplasia, retinal dysplasia/pigmentary dystrophy, juvenile cataract, digit abnormalities, tooth enamel anomalies, oligodontia, and phalangoepiphyseal dysplasia. Other syndromes with hair abnormalities may also mimic uncombable hair syndrome clinically and these include, Rapp-Hodgkin ectodermal dysplasia; loose anagen hair syndrome; ectodermal dysplasia, ectrodatyly, cleft lip/palate (EEC) syndrome; and familial tricho-odonto-onchyial ectodermal dysplasia with syndactyly. Unlike other conditions with an uncombable hair component, uncombable hair syndrome alone (cheveux incoiffables, pili trianguli et canaliculi) is not associated with physical, neurologic, or mental abnormalities. In most cases of uncombable hair syndrome, the hair is grossly abnormal in infancy and early childhood, but may have improved manageability later in life. Scanning electron microscopy of hair samples provides definitive evidence for diagnosis of clinically suspected uncombable hair syndrome and eliminates other hair abnormalities from the differential diagnosis. PMID- 11407535 TI - Metabolism of styrene in the human liver in vitro: interindividual variation and enantioselectivity. AB - 1. The interindividual variation and enantioselectivity of the in vitro styrene oxidation by cytochrome P450 have been investigated in 20 human microsomal liver samples. Liver samples were genotyped for the CYP2E1*6 and CYP2E1*5B alleles. 2. Kinetic analysis indicated the presence of at least two forms of styrene metabolizing cytochrome P450. The enzyme constants for the high-affinity component were subject to appreciable interindividual variation, i.e. Vmax1 ranged from 0.39 to 3.20 nmol mg protein(-1) min(-1) (0.96+/-0.63) and Km1 ranged from 0.005 to 0.03 mM (0.011+/-0.006). Inhibition studies with chemical inhibitors of CYP2E1, CYP1A2, CYP2C8/9 and CYP3A4 demonstrated that CYP2E1 was the primary enzyme involved in the high-affinity component of styrene oxidation. No relationship between the interindividual variation in Vmax1 and Km1 and the genetic polymorphisms of the CYP2E1 gene was found. 3. Cytochrome P450-mediated oxidation of styrene demonstrated a moderate enantioselectivity, with an enantiomeric excess (ee) of (S)-styrene oxide of 15% (range 4-27%) at low styrene concentration and an ee of (R)-styrene oxide of 7% (range -11 to +22%) at high styrene concentration. This points towards the involvement of at least two cytochrome P450, with different enantioselectivities. 4. The data indicate that cytochrome P450-mediated styrene oxidation is subject to considerable interindividual variation, but only to a moderate product enantioselectivity. PMID- 11407536 TI - Species difference in stereoselective involvement of CYP3A in the mono-N dealkylation of disopyramide. AB - 1. To determine which CYP isoenzyme is involved in the N-dealkylation of disopyramide (DP) metabolism in human and dog, and to determine the stereoselectivity of DP metabolism with human CYP and dog CYP isoenzymes, the following in vitro metabolism studies of DP were conducted: correlation between human CYP isoenzyme activities and DP metabolism with human liver microsomes; inhibition of DP metabolism in human and dog liver microsomes with chemical inhibitors of CYP isoenzymes; inhibition of DP metabolism in human microsomes with human CYP antibodies; inhibition of DP metabolism in dog liver microsomes with human and dog CYP antibodies; metabolism of DP with human (CYP3A4) and dog (CYP3A12) cDNA-expressed isoenzymes; determination of Km and Vmax of DP enantiomers by using cDNA-expressed CYP3A4 and CYP3A12. 2. In human liver microsomes, the formation of the mono-N-dealkylated disopyramide (MNDP) metabolite was best correlated with CYP3A4 activities. DP metabolism was substantially inhibited by ketoconazole, troleandomycin (TA) and human CYP3A4 antibody. DP was metabolized by cDNA-expressed CYP3A isoenzymes. In dog liver microsomes, DP metabolism was inhibited by ketoconazole, TA and dog anti-CYP3A12. DP was also metabolized by cDNA-expressed CYP3A12. 3. CYP3A4 and CYP3A12 are the principal isoenzymes involved in DP metabolism in human and dog respectively. There was no stereoselectivity in N-dealkylation of DP by human CYP3A4. However, there was notable stereoselectivity in the N-dealkylation by dog CYP3A12. PMID- 11407537 TI - PCB methyl sulphones in rat liver after exposure to PCB (Clophen A50): analysis and radiosynthesis of selected methylsulphonyl-PCBs. AB - 1. The hepatic metabolism of polychlorinated biphenyls (PCBs) and formation of PCB methyl sulphone metabolites (MeSO2-PCBs) was determined in the male Sprague Dawley rat 1, 2, 4 or 8 weeks after dosage with Clophen A50 (a commercial PCB mixture). 2. The total concentration of the PCB congeners examined (sigmaPCB) decreased during the experimental period, from 40 microg g(-1) lipid (l.w.) after 1 week to 4 microg g(-1) l.w. after 8 weeks. A 50% decrease of PCB in the liver were estimated to be 28, 13 and 11 days for 2,2',4,4',5,5'-hexachlorobiphenyl (CB153), 2,2',3,3',4,4',5-heptaCB (CB170) and 2,2'3,4',5,5',6-heptaCB (CB187), respectively. 3. The total MeSO2-PCB (sigmaMeSO2-PCB) concentration increased from 800 to 1,020 ng g(-1) l.w. during the first 2 weeks of treatment and thereafter a decrease to 120 ng g(-1) l.w. after 8 weeks. The relative concentration of both 3'-MeSO2-2,2',4,4',5-pentaCB (3'-MeSO2-CB101) and 3'-MeSO2 2,2',3,4,5'-pentaCB (3'-MeSO2-CB87) in rat liver showed significant increases during the 8 weeks. In contrast, the relative concentrations of 4-MeSO2-2,4',5,5 tetraCB (4-MeSO2-CB64), 3-MeSO2-2,3',4',5-tetraCB (3-MeSO2-CB70) and 4-MeSO2 2,2',3,4,5',6'-hexaCB (4'-MeSO2-CB132) decreased significantly. 4. A route for the synthesis of radiolabelled MeSO2-PCBs was developed employing the 'Pummerer reaction' to convert methylthio-PCBs (MeS-PCBs) to the corresponding mercapto PCBs (SH-PCB). The SH-PCBs were methylated with radiolabelled methyl iodide and the resulting sulphides oxidized to yield the corresponding MeSO2-PCBs. Using this approach, 3-[14C]-MeSO2-2,2',4',5,5',6-hexaCB (3-[14C]-MeSO2-CB149), 4-[14C] MeSO2-2,2',4',5,5',6-hexaCB (4-[14C]-MeSO,-CB149), 3'-[14C]-MeSO2-CB101, 4'-[14C] MeSO2-2,2',4,5,5'-pentaCB (4'-[14C]-MeSO2-CB101) and 4'-[3H]-MeSO2-CB101 were synthesized in quantitative radiochemical yields. PMID- 11407538 TI - Bioactivation to free radicals and cytotoxicity of 1,1-dichloro-1-fluoroethane (HCFC-141b). AB - 1. The in vitro bioactivation by rat liver microsomes and the cytotoxicity in rat hepatocytes of 1,1-dichloro-1-fluoroethane (HCFC-141b), a replacement for some ozone depleting chlorofluorocarbons (CFC), have been investigated. 2. Anaerobic incubations of liver microsomes from pyridine-induced rats with HCFC-141b in the presence of the spin-trapping agent N-t-butyl-alpha-phenylnitrone (PBN) resulted in the formation of a typical ESR radical signal. 3. In the presence of HCFC 141b, a dose-dependent formation of conjugated dienes was observed that was partially inhibited by PBN, glutathione (GSH) and vitamin C. Moreover, HCFC-141b increased the release of lactate dehydrogenase (LDH) and the depletion of cellular glutathione in isolated rat hepatocytes under both normoxic and hypoxic conditions. 4. HCFC-141b-dependent cytotoxicity was completely prevented by PBN under both conditions and it was partially prevented under normoxic conditions by the broad-spectrum P450 inhibitor metyrapone, the P4502E1 specific inhibitor 4 methylpyrazole and the P4503A-specific inhibitor troleandomycin. Interestingly, HCFC-141b-dependent glutathione depletion was not prevented by PBN, metyrapone, 4 methylpyrazole or troleandomycin, whereas two glutathione depletors, 2,6-dimethyl 2,5-heptadien-4-one (phorone) and diethylmaleate, partially prevented LDH release. 5. The present results indicate that HCFC-141b is reductively metabolized in vitro to free radical intermediates by P450, in particular by the CYP2E1 and, to a lower extent, CYP3A isoforms, leading to peroxidative membrane damage and glutathione-independent cytotoxicity. PMID- 11407539 TI - The management of VTEC O157 infection. AB - VTEC O157 infections, although showing a relentless rise in incidence over the last decade, only account for less than 10% of total food poisoning notifications in the UK. Despite this, the propensity for this infection to cause the serious and life-threatening clinical complications of haemolytic uraemic syndrome (HUS) and thrombotic thrombocytopaenic purpura (TTP), in a significant proportion (2 15%) of sufferers, highlights the need to focus on it both epidemiologically and clinically. The mortality rate of these complications (3-17% and up to 30% in outbreaks) adds urgency to this consideration. The pathogenesis and epidemiology of the illness caused by VTEC O157 is now well described, allowing the potential for appropriate intervention in outbreak and individual clinical management. The presence or absence of symptoms, e.g. bloody diarrhoea, fever, vomiting, in VTEC O157 infections compared with other causes of gastroenteritis may allow some selection of cases for more intensive management. Age (< 15, > 65 years), clinical hypochlorhydria. and a short incubation period have been associated with complication (HUS/TTP) development. Antibiotic therapy in the pre-infection period may predispose to complication development and there is evidence that it may increase complications if used in the management of acute illness. Laboratory markers such as early neutrophil leukocytosis have been shown both to correlate with VTEC O157 infection and to predict complications in central Scotland and Japan. The serum albumin and the C-reactive protein may act as additional markers for HUS development. Laboratory markers may be differentiated into those predicting HUS/TTP and those useful in monitoring its development. A scheme for clinical management of affected cases is presented to allow the attending clinician to select cases that may benefit from further intervention to prevent or treat complications. PMID- 11407540 TI - The fate of Escherichia coli O157 in soil and its potential to contaminate drinking water. AB - The survival and transport of Escherichia coli and E. coli O157 after cattle slurry application were studied on drained plots in both grassland and arable stubble at three sites in Scotland. Leaching losses were between 0.2% and 10% of total E. coli and were dependent on rainfall. Recovery of E. coli in grass and soil declined with approximately first order kinetics. Residual numbers, in excess of background declined more slowly. The pattern was similar for both grass and arable plots. Laboratory incubations of soil cores, with applied slurry containing E. coli and E. coli O157 were performed in soils with different moisture contents at two temperatures for clay loam and sandy loam soils. Both E. coli populations were measured over a 4-week period. Using a dual population approach, the die off of the susceptible pool was linear with a half-life of 3-4 days, and was faster at the higher temperature and lowest moisture content. The resistant pool was not strongly affected by temperature or moisture and had a half-life for die off of between 18 and 24 days. After a 4-week period, < 100 cfu g/soil of E. coli and E. coli O157 remained. The die off rate of E. coli O157 was the same or slightly faster than that of the commensal E. coli population, indicating that the field behaviour of E. coli O157 can be studied by monitoring the total population of E. coli applied with slurry. The risk of significant pollution of water by E. coli is highest immediately after application of slurry, and the first increments of drainflow carry significant concentrations. Thereafter, the risk of pollution is very low. If weather conditions are dry after application on well-drained sandy soils, it is unlikely that any significant losses of organisms to drains will occur. Such data can be used to control and minimise the risk of E. coli O157 contaminating drinking water. PMID- 11407541 TI - Beef HACCP: intervention and non-intervention systems. AB - While there are several generic beef HACCP documents available to the beef industry, these lack sufficient detail to be of any use other than as a general guide to HACCP. A document which clearly identifies and provides a sound scientific basis for potential critical control points (CCPs') and details critical limits, monitoring and corrective actions is clearly required. The objective of this paper is to provide such information. A detailed description of CCPs' for two different HACCP systems (an intervention and a non-intervention system) are presented and the advantages and disadvantages of each are discussed. Individual beef plants may then make an informed choice as to which HACCP system is most suitable for them and have all the specific information required for effective implementation. PMID- 11407542 TI - Zero-tolerance for faecal contamination of carcasses as a tool in the control of O157 VTEC infections. AB - The Dutch government, the meat producers organisation and the meat industry have recognised O157 VTEC as an important public health hazard, and agreed on the necessity to improve the hygiene in Dutch cattle- and calf-slaughtering establishments. This paper reports activities within a national action programme to achieve this objective, "Zero-tolerance for faecal contamination during slaughter of cattle and calves". The study included inspection of hygienic performances in slaughterhouses, and visual and microbiological (aerobic plate counts, Enterobacteriaceae counts and O157 VTEC presence/absence on visually clean cattle and calf carcasses) assessment of carcass cleanliness. Initial studies concluded that the hygienic performances in the Dutch cattle and calf slaughterhouses should be immediately improved. In 52% of the slaughterhouses inspected, carcasses were observed to be contaminated with hide, hair or faeces. Around 45% of the slaughterhouses had constructural deficiencies likely to lead to structural cross-contamination of carcasses, by direct carcass-carcass contact, or by indirect contacts with floors, walls or steps. In 39% of the slaughterhouses, cleaning and disinfection procedures were inadequate. Visual inspection of chilled carcasses found that in 11 of the 27 slaughterhouses visited, more than 10% of the carcasses were visibly contaminated. In 6 of the 27 slaughterhouses visited, more than 50% of the carcasses inspected were visibly contaminated. Microbiological analysis of visually clean carcasses noted contamination levels similar to those reported from other countries. O157 VTEC were not isolated during this study. Circulation of these findings lead to increased efforts by all parties to fulfil the requirements of the statutory "Zero-tolerance" programme. A follow-up study noted a significant decrease in the proportions of faecally contaminated carcasses, i.e., 7% of chilled carcasses were visibly contaminated with faeces, as opposed to 22% contamination during the initial study. The follow-up study also noted a greater awareness of the importance of good hygienic practices among slaughterhouse personnel and government meat inspectors. PMID- 11407543 TI - Quantitative microbiological risk assessment. AB - The production of safe food is being increasingly based on the use of risk analysis, and this process is now in use to establish national and international food safety objectives. It is also being used more frequently to guarantee that safety objectives are met and that such guarantees are achieved in a cost effective manner. One part of the overall risk analysis procedure-risk assessment is the scientific process in which the hazards and risk factors are identified, and the risk estimate or risk profile is determined. Risk assessment is an especially important tool for governments when food safety objectives have to be developed in the case of 'new' contaminants in known products or known contaminants causing trouble in 'new' products. Risk assessment is also an important approach for food companies (i) during product development, (ii) during (hygienic) process optimalization, and (iii) as an extension (validation) of the more qualitative HACCP-plan. This paper discusses these two different types of risk assessment, and uses probability distribution functions to assess the risks posed by Escherichia coli O157:H7 in each case. Such approaches are essential elements of risk management, as they draw on all available information to derive accurate and realistic estimations of the risk posed. The paper also discusses the potential of scenario-analysis in simulating the impact of different or modified risk factors during the consideration of new or improved control measures. PMID- 11407544 TI - Survival of Escherichia coli O157:H7 during the fermentation of Spanish-style green table olives (conservolea variety) supplemented with different carbon sources. AB - Olives were supplemented with two different carbon sources (sucrose and glucose) at five different levels (0%, 0.1%, 0.3%, 0.5% and 1%) and the fermentation procedure was followed after inoculation with or without starter culture. Inhibition of Escherichia coli O157:H7 was evident in all fermentation procedures regardless of the treatment used. The pathogen numbers declined but did not die out during fermentation. It was evident that the rate of death was higher in samples supplemented with starter cultures compared to natural fermentation. The production of acids during fermentation seems to be the main factor that governs the behaviour of this pathogen under such stress conditions. The HPLC analysis revealed that lactate, formate and acetate were among the end-products during the fermentation of green olives. PMID- 11407545 TI - Effect of stress induced by suboptimal growth factors on survival of Escherichia coli O157:H7. AB - This study investigated the growth and survival of E. coli O157:H7 exposed to a combination of suboptimal factors (22 degrees C, 7 degrees C, -18 degrees C/0.5% NaCl, 5.0% NaCl/pH 7.0, pH 5.4, pH 4.5/addition of lactic acid) in a simulation medium for red meat (beef gravy). Prolonged survival was noted as the imposed stress was more severe, and as multiple growth factors became suboptimal. At a defined temperature (7 degrees C or -18 degrees C), survival was prolonged at the more acid, more suboptimal pH (pH 4.5 > pH 5.4 > pH 7.0) while at a defined pH (pH 4.5), better survival was observed at 7 degrees C than at 22 degrees C. This suggests that application of the hurdle concept for preservation of food may inhibit outgrowth but induce prolonged survival of E. coli O157:H7 in minimal processed foods. At both 22 degrees C and 7 degrees C, the addition of lactic acid instead of HCl to reduce pH (to pH 4.5) resulted in a more rapid decrease of E. coli O157:H7. High survival was observed in beef gravy, pH 5.4 at -18 degrees C (simulation of frozen meat)-reduction of log 3.0 to log 1.9 after 43 days--and in beef gravy, pH 4.5 and 5% NaCl at 7 degrees C (simulation of a fermented dried meat product kept in refrigeration)--less than 1 log reduction in 43 days. In these circumstances, however, a high degree of sublethal damage of the bacterial cells was noted. The degree of sublethal damage can be estimated from the difference in recovery of the pathogen on the non-selective TSA medium and the selective SMAC medium. PMID- 11407546 TI - Use of strain typing to provide evidence for specific interventions in the transmission of VTEC O157 infections. AB - Transmission of verocytotoxin (VT)-producing Escherichia coli (VTEC) occurs by three main routes. These comprise food- or water-borne infections, acquisition of disease by direct or indirect contact with animals and person-to-person spread. Phenotypic typing of VTEC belonging to serogroup O157 is achieved by phage typing and identification of VT type. These properties quickly provide evidence for the linkage of human cases and their association to potential sources. DNA-based subtyping methods such as pulsed field gel electrophoresis (PFGE) are generally required to increase discrimination of VTEC O157 strains so that the spread of specific strains can be monitored. Phenotypic and DNA-based methods were used in the investigation of 85 general outbreaks of VTEC O157 infection between 1995 and 1999. Results were used in conjunction with epidemiological data to provide direct or indirect evidence for the likely route of transmission. Detailed strain fingerprinting identified specific food vehicles and reservoirs of infection in animals. Typing supported the implementation of measures to control the spread of infection that included pasteurisation orders, product withdrawal, temporary closure of retail premises and open farms and the introduction of HACCP-based working practices. In outbreaks involving widely distributed foods, DNA-based examination of apparently sporadic isolates with the same phage and VT type as outbreak strains was performed to identify additional potential outbreak cases and estimate the spread of infection. Strain typing was applied in outbreaks in nurseries and other institutions to monitor person-to-person spread, including careers and their families and to assess the involvement of community cases occurring at the same time. Rapid exchange of epidemiological, microbiological and typing data will be increasingly important in investigation of VTEC O157 outbreaks. PMID- 11407547 TI - Faecal carriage of Verocytotoxin-producing Escherichia coli O157 and carcass contamination in cattle at slaughter in northern Italy. AB - A study on the prevalence of the faecal carriage of Verocytotoxin (VT)-producing Escherichia coli (VTEC) O157 and on the rate of carcass contamination was carried out on feedlot cattle and dairy cows at slaughter in northern Italy. Between April 1998 and January 1999, 12 sampling visits were performed on different days in seven different slaughterhouses. At each visit, 5-12 animals consecutively slaughtered were selected. From each animal, faeces were collected from the rectum immediately after slaughter and surface swabs were taken from the leg region and the diaphragmatic insertion of the carcass. All samples were examined for the presence of VTEC O157 using an immunomagnetic separation technique. A total of 100 animals coming from 60 different farms were examined. In total, VTEC O157 was isolated from the intestinal content of 17, and from the carcasses of 12 of the 100 animals examined. In particular, VTEC O157 was recovered from six (35.3%) out of the 17 carcasses from which the organism had previously been isolated from rectal content and from six (7.3%) of the 82 carcasses of the stool negative cattle. In seven carcasses, VTEC O157 was isolated from the leg area, in two carcasses from the diaphragmatic area, and in three carcasses from both areas. Major differences in the prevalence of VTEC O157 were observed in the different groups of cattle sampled. In 7 of the 12 sampling visits, all the specimens examined were negative, while 16 of the 17 positive stool samples and 11 of the 12 positive carcass swabs were collected during three of the visits, performed in June in three different abattoirs. In these three visits, the ratios between the percentage of animals carrying VTEC O157 in the stools and the percentage of contaminated carcasses were 0.33, 0.57, and 1.66, respectively; thus, confirming that slaughter practices can largely influence the rate of carcass contamination. Phage typing and PFGE analysis of VTEC O157 isolated from samples collected at the same visit suggested that both auto- and cross contamination occurred. PMID- 11407548 TI - Persistence of verocytotoxin-producing Escherichia coli O157:H7 in calves kept on pasture and in calves kept indoors during the summer months in a Swedish dairy herd. AB - In 1997, a Swedish dairy farm was implicated in a human case of verocytotoxigenic Escherichia coli (VTEC) infection. The bacterium was found in a faecal sample from the human case and in faecal samples from cattle on the farm. Subtyping with pulsed field gel electrophoresis (PFGE) showed that the isolates were identical. The farm was further studied to assess the occurrence and the epidemiology of the agent at the farm level. The objective of this part of the study presented here was to examine the persistence of VTEC O157:H7 in calves that were kept on pasture and indoors, respectively, during the summer. Twelve calves in the herd, with one positive faecal sample each of VTEC O157:H7 in April 1999, were followed by faecal sampling during the summer months. Six calves were kept indoors and six were kept on pasture. Faecal samples from each calf were collected once a month on five occasions from April to September. Bacterial examination was performed with immunomagnetic separation (IMS) and cultivation on CT-SMAC. PCR was used to test for the presence of genes encoding for verocytotoxin (VT), intimin (eaeA), enterohemorrhagic E. coli-hemolysin (EHEC-Hly) and the flagellar antigen H7. PFGE was used for genotyping the isolates. The faecal samples from the calves kept on pasture were negative during the whole period. It is possible that the faecal samples had bacterial counts lower than the detection limits for our procedure, or that the faecal samples were free from the bacteria at the time of sampling. This suggests that calves on pasture may be less exposed to the bacteria or that they clear themselves. In the pen group, there were between one and six culture positive individuals per sampling occasion. One of the calves that was housed indoors was positive in faecal culture on four consecutive samplings. PMID- 11407549 TI - A survey of the prevalence of Escherichia coli O157 in raw meats, raw cow's milk and raw-milk cheeses in south-east Scotland. AB - 2429 samples of foodstuffs were examined for the presence of verocytotoxigenic Escherichia coli O157 (VTEC O157) by means of immunomagnetic separation (IMS) over a 2-year period commencing April 1997. Specimens comprised 1190 raw meats, 500 raw milks and 739 raw-milk cheeses. The meat and cheese samples were purchased from retail premises in south-east Scotland; raw milk samples were obtained directly from farms. In addition, total E. coli counts were performed on milk and cheese samples, and the pH of cheese specimens measured. The water activity (Aw) was also measured for a representative sample of each cheese type, and for all of the samples with high levels of E. coli. VTEC O157 was isolated from two samples of beef burger, both manufactured on the premises of the same butchers shop. Control studies with artificially inoculated foodstuffs demonstrated a sensitivity of detection of < 5 organisms 25 g(-1). These findings, which contrast with the results of similar studies elsewhere in the UK, suggest that other sources of infection may be important in explaining the high rates of infection with this organism in south-east Scotland. PMID- 11407550 TI - The control of VTEC in the animal reservoir. AB - A great diversity of VTECs exist but only in the case of Escherichia coli O157:H7, a common human foodborne pathogen, has sufficient research been done to allow generalizations about the ecology. The key features are as follows: lack of host specificity such that indistinguishable isolates can be found in a variety of species; near-ubiquitous distribution in cattle (and perhaps other ruminant) farms; transient residence in the gastrointestinal flora of individual animals that is not associated with clinical disease; temporal clustering at the population level such that most fecal shedding is confined to sharp bursts in a high percentage of animals separated by much longer periods of very low prevalence; a higher prevalence in young animals in comparison to older ones: a higher prevalence in animals with floral disturbance such as that caused by transit, feed changes or antimicrobial dosing; and a markedly higher prevalence during warm months. Molecular epidemiological studies of E. coli O157:H7 have demonstrated that subtypes of the organism can persist on cattle farms for years, thus supporting a conclusion that cattle farms represent a reservoir. Yet on such farms, common subtypes are often found in environmental niches and in other species of animals; thus, it is not completely clear that cattle themselves are the reservoir. New subtypes are periodically observed on particular farms, and indistinguishable subtypes can be found on farms that are separated by hundreds of kilometers even in the absence of any obvious animal movements between them. The number of subtypes found on a farm does not appear to be qualitatively correlated with cattle movements (e.g., purchases) into the farm. Commercial feeds are sometimes contaminated with E. coli O157:H7, and it seems likely that feeds represent an important route of dissemination for this agent and other VTEC. Mixed feeds collected from feeding troughs are commonly positive for E. coli O157:H7, as are cattle watering troughs, and feed and water likely represent the most common means of infection. Environmental replication in feeds and in sediments of watering troughs occurs and may account for the higher level of fecal shedding in the warm months. Since E. coli O157:H7 has been found to persist and remain infective for at least 6 months in water trough sediments, this may be an important environmental niche where the organism survives during periods when it cannot be detected in cattle, especially during cold months. Traditional means of controlling infectious agents, such as eradication or test and removal of carrier animals, do not appear to be feasible for VTECs. Nevertheless, certain farm management practices-especially those related to maintenance and multiplication of the agent in feed and water-may provide practical means to substantially reduce the prevalence of these agents in cattle on farms and in those arriving at slaughter plants. PMID- 11407551 TI - Control of VTEC in Dutch livestock and meat production. AB - The Dutch government and the meat industry, recognising VTEC as having important public health, meat quality and economic implications, have taken a number of initiatives within the last 5 years to control VTEC in livestock and meat. These initiatives, brought together last year in a 'Masterplan VTEC', include short-, middle- and long-term priorities. Short-term priorities include advice on interventions in the cases of an outbreak of VTEC associated with a cattle herd, the implementation of handbooks for Good Manufacturing Practice (GMP) in slaughterhouses and deboning plants, and the execution of an action programme on zero-tolerance to faecal contamination of carcasses. Mid-term activities include surveillance of the occurrence of VTEC and other enteropathogens in livestock and meat, and the investigations of VTEC population dynamics in dairy farms, transportation and farm hygiene. In the longer term, this programme aims to produce a system of Integrated Quality Assurance, consolidating effective measures to control VTEC in Dutch livestock and meat, and integrating emerging means for control and prevention. PMID- 11407552 TI - A field survey of Escherichia coli O157 ecology on a cattle farm in Italy. AB - A field survey was performed in a heifer raising operation in Northern Italy to study the introduction, maintenance and dissemination of Escherichia coli O157 in the herd and to identify possible control measures at the farm level. Rectal swabs from two different groups of animals (surveys 1 and 2) were tested for E. coli O157 by an immunomagnetic separation technique. In survey 1, a group of female calves (341 animals initially) introduced from 30 dairy herds during April 1996 to March 1997 were tested for E. coli O157 on arrival from the original herd when housed in individual hutches, 2-3 days after completion of weaning (which was associated with grouping) and 2 months after weaning. No statistically significant difference between excretion rates (3.8%, 4.2%, 4.4%, respectively) was found. Calves from which E. coli O157 was isolated on arrival came from 6 of the 30 dairy herds. Strains isolated during survey 1 belonged to seven different pulsed field gel electrophoresis (PFGE) profiles. In survey 2, a group of young animals aged, at the beginning of the study, between 2 1/2 and 7 1/2 months (median = 124 days) was tested monthly for E. coli O157 for 11-15 months from May 1996 to July 1997. The group included 92 animals for 11 months and then gradually decreased to 59 animals. Overall, E. coli O157, belonging to six different PFGE profiles, were isolated from 138 (10.7%) of 1293 rectal swabs. Monthly excretion rates ranged from 2.7% to 23.7%, with summer peaks in both years. Fifty-nine (64.1%) of the 92 heifers were positive at least once: of these 59 animals, 22 (37.3%) were positive on only one occasion, 23 (39%) were positive on two occasions and 14 (23.7%) were positive on three or more occasions. From two heifers positive on 9 out of the 15 sampling visits, strains with the same PFGE profile were isolated, respectively, on seven and eight occasions while strains with only one band difference were isolated on the remaining occasions. E. coli O157 was also isolated from 6 of 16 samples of bedding, two of two samples of slurry and one of five samples from water troughs collected during survey 2. PMID- 11407553 TI - Occurrence of Shiga toxin-producing Escherichia coli in fecal samples from children with diarrhea and from healthy zebu cattle in Uganda. AB - Fecal samples collected from 237 diarrheic infants in Kampala, Uganda and from 159 healthy cattle from a ranch in the Central Region of Uganda were investigated for the presence of Escherichia coli O157 and other types of Shiga toxin producing E. coli (STEC). E. coli O157 were not detected in 150 stool samples from children which were cultivated on sorbitol MacConkey agar. A search for all types of STEC performed on 87 additional human stool samples with an enzyme immunoassay for Shiga toxins (Premier EHEC) was also negative. Forty-two stool samples from infants were additionally investigated for enteropathogenic E. coli (EPEC) by DNA-hybridization with an eae-specific gene probe. Compared to STEC, EPEC were frequent and found in six (14.3%) of these 42 randomly selected stool specimens. We were further interested in the role of cattle as a reservoir for STEC in Uganda. STEC were isolated from 45 of 159 cattle from a herd in the Central Region of Uganda. STEC strains from cattle belonged to 16 different O- and nine different H-types and nine O:H types were identical to those found in bovine STEC from other continents. Only one bovine STEC strain was positive for the eae-gene, and O-groups associated with enterohemorrhagic E. coli (EHEC) types (O26, O103, O111, O145 and O157) were not found. Our report demonstrates that STEC are not frequent in urban children in Uganda, but domestic cattle were identified as an important natural reservoir for these organisms in this country. PMID- 11407554 TI - Hearing-aid outcome measured following one month of hearing aid use by the elderly. AB - This study reports the results of a large number of hearing-aid outcome measures obtained from 173 elderly hearing-aid wearers following one month of hearing-aid use. All participants in this study were fit binaurally with identical full concha in-the-ear (ITE) hearing aids having linear Class-D amplifiers with output limiting compression. Outcome measures included several measures of speech recognition, as well as several self-report measures of hearing-aid performance, benefit, satisfaction, and use. Comparison of mean data from this sample of hearing-aid wearers to other larger sets of data, obtained previously for several of these measures of hearing-aid outcome evaluated in isolation, indicated that the participants in this study were representative of the participants in other larger-scale studies. Subsequent principal-components factor analysis of the data from this study indicated that there were seven distinct dimensions of hearing aid outcome. Attempts to document the effectiveness and efficacy of hearing aids for elderly persons with impaired hearing will be most complete when assessing performance along all seven dimensions of hearing-aid outcome. Clinically efficient procedures for doing so are discussed. PMID- 11407555 TI - Recognition of speech produced in noise. AB - A two-part study examined recognition of speech produced in quiet and in noise by normal hearing adults. In Part I 5 women produced 50 sentences consisting of an ambiguous carrier phrase followed by a unique target word. These sentences were spoken in three environments: quiet, wide band noise (WBN), and meaningful multi talker babble (MMB). The WBN and MMB competitors were presented through insert earphones at 80 dB SPL. For each talker, the mean vocal level, long-term average speech spectra, and mean word duration were calculated for the 50 target words produced in each speaking environment. Compared to quiet, the vocal levels produced in WBN and MMB increased an average of 14.5 dB. The increase in vocal level was characterized by increased spectral energy in the high frequencies. Word duration also increased an average of 77 ms in WBN and MMB relative to the quiet condition. In Part II, the sentences produced by one of the 5 talkers were presented to 30 adults in the presence of multi-talker babble under two conditions. Recognition was evaluated for each condition. In the first condition, the sentences produced in quiet and in noise were presented at equal signal-to noise ratios (SNR(E)). This served to remove the vocal level differences between the speech samples. In the second condition, the vocal level differences were preserved (SNR(P)). For the SNR(E) condition, recognition of the speech produced in WBN and MMB was on average 15% higher than that for the speech produced in quiet. For the SNR(P) condition, recognition increased an average of 69% for these same speech samples relative to speech produced in quiet. In general, correlational analyses failed to show a direct relation between the acoustic properties measured in Part I and the recognition measures in Part II. PMID- 11407556 TI - Effects of linguistic cues and stimulus cohesion on intelligibility of severely dysarthric speech. AB - This study examined the effects of supplemental cues on the intelligibility of unrelated sentences and related sentences (narratives) produced by 4 women with severe dysarthria secondary to cerebral palsy. Visual images containing alphabet, topic, combined (alphabet and topic together), and no cues were imposed in real time on audio speech samples and presented to 72 nondisabled listeners. Statistical results showed that cue conditions had similar effects on unrelated and on related sentence intelligibility. Combined cues resulted in higher intelligibility scores than any other cue condition, no cues resulted in lower intelligibility scores than any other cue condition, and alphabet cues yielded higher intelligibility scores than topic cues. Intelligibility of related and unrelated sentences differed only for alphabet cues where related sentences had greater intelligibility than unrelated sentences. Results are discussed relative to the quantity and type of cues. PMID- 11407557 TI - Voice activity and participation profile: assessing the impact of voice disorders on daily activities. AB - Traditional clinical voice evaluation focuses primarily on the severity of voice impairment, with little emphasis on the impact of voice disorders on the individual's quality of life. This study reports the development of a 28-item assessment tool that evaluates the perception of voice problem, activity limitation, and participation restriction using the International Classification of Impairments, Disabilities and Handicaps-2 Beta-1 concept (World Health Organization, 1997). The questionnaire was administered to 40 subjects with dysphonia and 40 control subjects with normal voices. Results showed that the dysphonic group reported significantly more severe voice problems, limitation in daily voice activities, and restricted participation in these activities than the control group. The study also showed that the perception of a voice problem by the dysphonic subjects correlated positively with the perception of limitation in voice activities and restricted participation. However, the self-perceived voice problem had little correlation with the degree of voice-quality impairment measured acoustically and perceptually by speech pathologists. The data also showed that the aggregate scores of activity limitation and participation restriction were positively correlated, and the extent of activity limitation and participation restriction was similar in all except the job area. These findings highlight the importance of identifying and quantifying the impact of dysphonia on the individual's quality of life in the clinical management of voice disorders. PMID- 11407558 TI - The intelligibility of time-domain-edited esophageal speech. AB - The intelligibility of esophageal speech has been shown to be significantly lower than that of normal laryngeal speech. The current study investigated the possibility of enhancing the intelligibility of esophageal speech by manipulating samples in the time domain. Specifically, injection noises and nonphrasal pauses were digitally edited from the speech samples of 5 esophageal talkers. Twenty five sentences were selected and edited in the time domain and presented to 15 naive listeners who were instructed to write down the words that they heard. The percentage of correct words heard for each sentence was determined and compared across listeners, sentences, and talkers. The overall effect of the editing was a small but significant gain in the intelligibility of the esophageal speech. The improvement in intelligibility, however, depended on the individual talker, the speech material, and the number of editing changes made to a particular sample. PMID- 11407559 TI - Clinicoanatomic studies in dysarthria: review, critique, and directions for research. AB - More than 30 years ago, Darley, Aronson, and Brown (1969) proposed clinicoanatomic correlations for seven perceptual types of dysarthria. These correlations have not been systematically re-examined even though imaging technologies developed in recent years provide the means to do so. This review considers data from published imaging studies as well as data from selected medical interventions to evaluate the current state of knowledge that relates lesion site to the nature of a speech disturbance. Although the extant data are not sufficient to allow a complete evaluation of the seven types of dysarthria described by Darley et al., relevant information has been reported on lesions of the pyramidal pathway, extrapyramidal pathway, and cerebellum. In general, the results are best explained by an equivalence mode of brain-behavior relationship in which a type of dysarthria is associated with a lesion in one of two or more brain structures. Criteria also are proposed for future studies of clinicoanatomic relationships in neurogenic communication disorders. PMID- 11407560 TI - The effects of changes in hearing status in cochlear implant users on the acoustic vowel space and CV coarticulation. AB - In order to examine the role of hearing status in controlling coarticulation, eight English vowels in /bVt/ and /dVt/ syllables, embedded in a carrier phrase, were elicited from 7 postlingually deafened adults and 2 speakers with normal hearing. The deaf adults served in repeated recording sessions both before and up to a year after they received cochlear implants and their speech processors were turned on. Each of the two hearing control speakers served in two recording sessions, separated by about 3 months. Measures were made of second formant frequency at obstruent release and at 25 ms intervals until the final obstruent. An index of coarticulation, based on the ratio of F2 at vowel onset to F2 at midvowel target, was computed. Changes in the amount of coarticulation after the change in hearing status were small and nonsystematic for the /bVt/ syllables; those for the /dVt/ syllables averaged a 3% increase--within the range of reliability measures for the 2 hearing control speakers. Locus equations (F2 at vowel onset vs. F2 at vowel midpoint) and ratios of F2 onsets in point vowels were also calculated. Like the index of coarticulation, these measures tended to confirm that hearing status had little if any effect on coarticulation in the deaf speakers, consistent with the hypothesis that hearing does not play a direct role in regulating anticipatory coarticulation in adulthood. With the restoration of some hearing, 2 implant users significantly increased the average spacing between vowels in the formant plane, whereas the remaining 5 decreased that measure. All speakers but one also reduced vowel duration significantly. Four of the speakers reduced dispersion of vowel formant values around vowel midpoint means, but the other 3 did not show this effect. PMID- 11407561 TI - The effects of contextualization on fluency in three groups of children. AB - This study investigated the effects of contextualization on fluency in 12 school age children who stutter (CWS), 11 children with language impairment (CLI), and 12 children with normally developing fluency skills (CNF). Participants in the study were between the ages of 8 and 12 years and were matched for age and sex. Four discourse samples were elicited by asking participants to (a) generate two scripts related to cooking and (b) retell two stories. Having objects or pictures immediately available contextualized a cooking task and a retelling task; another set of cooking and retelling tasks were decontextualized. Moments of disfluency were identified and coded for three primary categories of disfluency: stuttering type, normal-type, and mazing. For CWS, a significant reduction in frequency of stuttering was noted in the contextualized script generation, and mazing occurred at a significantly higher frequency than did stuttering-type or normal-type disfluencies across the four tasks. For all three groups, both decontextualized conditions produced greater frequencies of normal-type disfluency and mazing. In addition, narrative retelling tasks yielded higher frequencies of disfluency than did the two cooking scripts. PMID- 11407562 TI - Effects of frequency-shifted auditory feedback on fundamental frequency of long stressed and unstressed syllables. AB - Twenty-four normally speaking subjects had to utter the test word /tatatas/ with different stress patterns repeatedly. Auditory feedback was provided by head phones and was shifted downwards in frequency during randomly selected trials while the subjects were speaking the complete test word. If the first syllable was long stressed, fundamental frequency of the vowel significantly increased by 2 Hz (corresponding to 25.5 cents) under frequency-shifted auditory feedback of .5 octave downwards, whereas under a shift of one semitone downwards a trend of an increase could be observed. If the first syllable was unstressed, fundamental frequency remained unaffected. Regarding the second syllable, significant increases or a trend for an increase of fundamental frequency was found in both shifting conditions. Results indicate a negative feedback mechanism that controls the fundamental frequency via auditory feedback in speech production. However, within a syllable a response could be found only if the syllable duration was long enough. Compensation for frequency-shifted auditory feedback still is quite imperfect. It is concluded that control of fundamental frequency is rather important on a suprasegmental level. PMID- 11407563 TI - What is stuttering? PMID- 11407564 TI - Ambiguity and algorithms in diagnosing early stuttering: comments on Ambrose and Yairi (1999). PMID- 11407565 TI - Word frequencies in toddlers' lexicons. AB - Word frequencies in toddlers' lexicons were examined in two studies using the Language Development Survey (LDS), a vocabulary checklist completed by parents (Rescorla, 1989). In Study 1, a high degree of consistency in LDS word frequencies was found when six samples of 24-month-olds were compared (total N = 758). Word frequency correlations in the .90s were found between large, unselected samples of toddlers of roughly similar socioeconomic status (SES). Correlations were somewhat lower but still highly significant when groups varying widely in SES were compared. In Study 2, LDS word frequencies in a sample of 40 late talkers traced from age 2 to 3 were compared to those in a large community sample from Study 1. Both lexicon size and age of the late talker influenced the degree of consistency found with respect to the community sample. The most common words reported in the lexicons of a sample of 422 24-month-olds were consistent with those identified in diary studies as among the highest frequency words used by young children in their early vocabularies. PMID- 11407566 TI - The effect of target-selection strategy on phonological learning. AB - In this study, 48 children with moderate or severe delays in phonological ability received treatment for four phonemes, selected in accordance with either traditional or nontraditional target-selection criteria. Children who received treatment for phonemes that are early developing and associated with greater productive phonological knowledge showed greater progress toward acquisition of the target sounds than did children who received treatment for late-developing phonemes that were associated with little or no productive phonological knowledge. Between-group differences in generalization learning were not observed. Child enjoyment of therapy did not differ between groups, but parental satisfaction with treatment progress was greater for children in the traditional group than for children in the nontraditional group. PMID- 11407567 TI - Training volunteers as conversation partners using "Supported Conversation for Adults with Aphasia" (SCA): a controlled trial. AB - This article reports the development and evaluation of a new intervention termed "Supported Conversation for Adults with Aphasia" (SCA). The approach is based on the idea that the inherent competence of people with aphasia can be revealed through the skill of a conversation partner. The intervention approach was developed at a community-based aphasia center where volunteers interact with individuals with chronic aphasia and their families. The experimental study was designed to test whether training improves the conversational skills of volunteers, and, if so, whether the improvements affect the communication of their conversation partners with aphasia. Twenty volunteers received SCA training, and 20 control volunteers were merely exposed to people with aphasia. Comparisons between the groups' scores on a Measure of Supported Conversation for Adults with Aphasia provide support for the efficacy of SCA. Trained volunteers scored significantly higher than untrained volunteers on ratings of acknowledging competence [F(1, 36) = 19. 1, p < .001] and revealing competence [F(1, 36) = 159.0, p < .001] of their partners with aphasia. The training also produced a positive change in ratings of social [F(1, 36) = 5.7, p < .023] and message exchange skills [F(1, 36) = 17.6, p < .001 ] of individuals with aphasia, even though these individuals did not participate in the training. Implications for the treatment of aphasia and an argument for a social model of intervention are discussed. PMID- 11407568 TI - Predictive inferencing in adults with right hemisphere brain damage. AB - Predictive inferencing was evaluated in 13 adults with right hemisphere damage (RHD) and 11 adults without brain damage (NBD). Brief narrative stimuli that strongly suggested a single outcome were constructed to vary recency of mention of inference-related information. Reading times were recorded for narrative-final sentences that disconfirmed the target inferences. Slowed reading time on the final sentences was an indicator of inference generation. Adults with RHD generated target predictive inferences in contexts with recent mention of strongly biasing inference-related information. This group also evidenced maintenance of inferences over time, but to a lesser degree than participants in the NBD group. Overall, individuals with better auditory comprehension or larger estimated working memory capacity tended to maintain inferences better than did the other participants. The results are discussed in relation to current hypotheses of inferencing and discourse comprehension in adults with RHD. PMID- 11407569 TI - Detection of irregular verb violations by children with and without SLI. AB - Fifty-seven children (ages in years;months: 5;7-8;8) with and without Specific Language Impairment (SLI) participated in judgment and elicitation tasks designed to evaluate their understanding of restrictions associated with irregular verb forms. The performance of the SLI group was similar to the performances of the control groups in that all children demonstrated high levels of sensitivity to violations involving verb-agreement errors (e.g., he am falling). The production and acceptance rates of past tense overregularizations (e.g., he falled) by the SLI and language-match groups were similar, and both were higher than the age match group. Differences between affected and unaffected children were observed in their productions and relative levels of sensitivity to infinitive errors in finite positions (e.g., he fall off). As expected, children in the SLI group were more likely to produce and accept infinitive forms in finite positions. Children in the SLI group also accepted more finite form errors in VP complement positions (e.g., he made him fell) than the control groups, although the latter occurred rarely in children's productions. Implications for understanding morphophonological and morphosyntactic development in children with SLI are discussed. PMID- 11407570 TI - Early phonetic and lexical development: a productivity approach. AB - Researchers frequently examine the development of the single-word lexicon in the absence of phonetic data. Yet a large body of literature demonstrates relationships between the phonetics of babble and early speech, and it is clear that production skill is essential for establishing a lexicon. This study uses longitudinal productivity criteria to establish children's phonetic skill. Twenty children were followed from age 9 to 16 months, and their level of consistency of vocal patterns was examined in relation to their lexical production, providing a relatively large-sample demonstration of phonetic/lexical relationships at the transition to language. Number of specific consonants produced consistently across the months of observation predicted referential lexical use at 16 months, whereas the transition to reference itself signaled the onset of a sharp increase in numbers of different words produced in a session. The earliest referential speakers exhibited prior consistency in the production of [p/b], which also predominated in their words. Prior use of at least two supraglottal consonants characterized the referential group. Children varied in the specific consonants they produced consistently, and these same consonants, varying according to individual child repertoire, characterized nearly all consonant-based words produced by each child in both of the final 2 months of observation. These findings are interpreted in relation to the children's contemporaneous development of representational ability and pragmatic skill. PMID- 11407571 TI - Psycholinguistic models of speech development and their application to clinical practice. AB - This article presents an introduction to psycholinguistic models of speech development. Two specific types of models are addressed: box-and-arrow models and connectionist or neural network models. We review some historical and some current models and discuss recent applications of such models to the management of speech impairment in children. We suggest that there are two ways in which a psycholinguistic approach can influence clinical practice: by directly supplementing a speech-language pathologist's repertoire of assessment and treatment approaches and by offering a new way to conceptualize speech impairment in children. PMID- 11407572 TI - New synthetic ways for the preparation of high-performance liquid chromatography supports. AB - The latest developments and in particular important synthetic aspects for the preparation of modern HPLC supports are reviewed. In this context, the chemistry of inorganic supports based on silica, zirconia, titania or aluminum oxide as well as of organic supports based on poly(styrene-divinylbenzene), acrylates, methacrylates and other, more specialized polymers is covered. Special consideration is given to modern approaches such as sol-gel technology, molecular imprinting, perfusion chromatography, the preparation of monolithic separation media as well as to organic HPLC supports prepared by new polymer technologies such as ring-opening metathesis polymerization. Synthetic particularities relevant for the corresponding applications are outlined. PMID- 11407573 TI - Experimental studies of uncertainties associated with chromatographic techniques. AB - The paper describes experiments for the evaluation of uncertainties associated with a number of chromatographic parameters. Studies of the analysis of vitamins by HPLC illustrate the estimation of the uncertainties associated with experimental "input" parameters such as the detector wavelength, column temperature and mobile phase flow-rate. Experimental design techniques, which allow the efficient study a number of parameters simultaneously, are described. Multiple linear regression was used to fit response surfaces to the data. The resulting equations were used in the estimation of the uncertainties. Three approaches to uncertainty calculation were compared--Kragten's spreadsheet, symmetric spreadsheet and algebraic differentiation. In cases where non-linearity in the model was significant, agreement between the uncertainty estimates was poor as the spreadsheet approaches do not include second-order uncertainty terms. PMID- 11407574 TI - High-performance liquid chromatography--ToxPrint: chromatographic analysis with a novel (geno)toxicity detection. AB - In order to aid the monitoring of the overall quality of (surface) waters a new analytical approach has been developed, combining on-line solid-phase extraction, HPLC separation and effect-related detection. Compounds present in surface water or wastewater samples are extracted on-line with Oasis [poly(divinylbenzene-co-N vinylpyrrolidone)] material and directly fractionated by reversed-phase HPLC. The eluent of the total chromatogram is collected on a microtitre plate in fractions of 1 min each. After evaporation and re-dissolvation in a suitable solvent, the (geno)toxicity of the individual fractions before and after enzymatic activation with S9, is determined with the umu test. In this way, harmful compounds can be detected and localized in the HPLC-diode array detection trace even without their identity and exact concentration being known at that moment. The method was developed using two test compounds, 4-nitroquinoline-N-oxide and 2 aminoanthracene. Compounds with mutagenic properties comparable to those of the test compounds can be detected from 0.1 microg/l, which is a concentration relevant for surface waters. The new analytical approach was successfully applied to various types of model samples, as well as real wastewater. PMID- 11407575 TI - Normal-phase high-performance liquid chromatographic separations using ethoxynonafluorobutane as hexane alternative. I. Analytical and chiral applications. AB - A novel, environmentally friendly, fluorinated solvent--ethoxynonafluorobutane- has been used to replace n-hexane in normal-phase HPLC applications. Fast gradients of methanol in ethoxynonafluorobutane on a cyano column have been successfully applied to the separation of steroids, benzodiazepines, NSAIDs, tricyclic antidepressants, beta-adrenergic blocking agents and mixtures of purines and pyrimidines. Small amounts of triethylamine and trifluoroacetic acid added to such gradients significantly improved peak shape and column performance for basic and acidic solutes. Ethoxynonafluorobutane and its mixtures with methanol have also been demonstrated to have a unique selectivity in chiral HPLC applications. PMID- 11407576 TI - Automated solid-phase extraction for sample preparation followed by high performance liquid chromatography with diode array and mass spectrometric detection for the analysis of resveratrol derivatives in wine. AB - A method has been developed for the simultaneous determination of resveratrol in all its forms (free isomers and glycosylates) in wines by high-performance liquid chromatography with diode array and mass spectrometric detection. Prior to injection into the column, preconcentration of the sample by automated solid phase extraction is carried out. In the detection by UV absorption, quantitation was carried out at 280 and 305 nm, and in detection by mass spectrometry, quantitation was performed in the selected ion monitoring mode at m/z 228 and at m/z 238. A comparative study between both detection systems was carried out. PMID- 11407577 TI - Urea gradient size-exclusion chromatography enhanced the yield of lysozyme refolding. AB - Protein refolding is still a bottleneck for large-scale production of valuable proteins expressed as inclusion bodies in Escherichia coli. Usually biologically active proteins cannot be obtained with high yield at a high concentration after refolding. In order to meet the challenge of protein refolding a urea gradient gel filtration-refolding system was developed in this article. A Superdex 75 column was pre-equilibrated with a linear decreased urea gradient, the denatured protein experienced the gradual decrease in urea concentration as it went through the column. The refolding of denatured lysozyme showed this method could significantly increase the activity recovery of denatured lysozyme at high protein concentration. The activity recovery of 90% was obtained from the initial protein concentration up to 17 mg/ml within 40 min. PMID- 11407578 TI - Chromatographic purification of some 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors. AB - The purification of pravastatin, simvastatin and lovastatin in the sodium salt or lactone form and of mevastatin in the lactone form by reversed-phase displacement chromatography is presented. The mobile phases consisted of water or mixtures of water-methanol and water-acetonitrile. Six different displacers were successfully used. Up to 0.14 g of raw sample per gram of stationary phase was loaded on a column packed with silica-based octadecyl phase. Crude substances from 85 to 88% chromatographic purity were purified and at least 99.5% purity was achieved. PMID- 11407579 TI - Simultaneous determination of six inorganic anions in drinking water by non suppressed ion chromatography. AB - A non-suppressed ion chromatographic method with conductometric detection is described for the simultaneous determination of six inorganic anions: fluoride, chloride, nitrite, bromide, nitrate and sulphate. The separation was achieved on a low-capacity anion-exchange column Metrohm IC Anion Column Super Sep, with a mobile phase consisting of phtalic acid dissolved in high-purity water, 2-amino-2 hydroxymethyl-1,3-propendiol and acetonitrile. In this work computer optimization procedures, using computer programs to select chromatographic conditions have been used, leading to the achievement of a desired separation. By using the different optimization methods in an integrated manner it is, however, possible to both speed method development, by reducing unnecessary experimentation, and to overcome the many shortcomings of each method, because of the different approaches. The purpose of this work is to improve and characterise the method for simultaneous determination of six inorganic anions in drinking water by non suppressed ion chromatography, using optimization procedures, in order to be applied to the routine analysis. The proposed method has numerous advantages over the other widely used non-suppressed ion chromatography methods: higher selectivity, shorter analysis time, lower quantitation and detection limits. The performance characteristics of the method were established by determining the following validation parameters: precision and accuracy, linearity, detection limits and quantitation limits. PMID- 11407580 TI - Determination of iodide in seawater and urine by size exclusion chromatography with iodine-starch complex. AB - A novel method for the determination of iodide by size exclusion chromatography was established. The method was simple and highly sensitive with good precision. Iodide was converted to iodine, then sequestered with starch, and separated from the matrix using a Shim-pack DIOL-150 (250 x 7.9 mm) size exclusion column with methanol-0.01 mol l(-1) aqueous phosphoric acid (10:90, v/v) as mobile phase at 1.2 ml min(-1) and UV detection at 224 nm. The calibration graph was linear from 1.0 ng ml(-1) to 100.0 ng ml(-1) for iodide with a correlation coefficient of 0.9992 (n=6). The detection limit was 0.2 ng ml(-1). The method was successfully applied to the determination of iodide in seawater and urine. The recovery was from 92% to 103% and the relative standard deviation was in the range of 1.5% to 3.7%. PMID- 11407581 TI - Analysis of steryl esters in cocoa butter by on-line liquid chromatography-gas chromatography. AB - On-line liquid chromatography-gas chromatography (LC-GC) has been applied to the analysis of steryl esters in cocoa butter. Separation of the steryl esters was achieved after on-line transfer to capillary GC. HPLC removes the large amount of triglycerides and pre-separates the components of interest, thus avoiding time consuming sample preparation prior to GC analysis. The identities of the compounds were confirmed by GC-MS investigation of the collected HPLC fraction and by comparison of the mass spectra (chemical ionization using ammonia as ionization gas) to those of synthesized reference compounds. Using cholesteryl laurate as internal standard, steryl esters were quantified in commercial cocoa butter samples, the detection limit being 3 mg/kg and the quantification limit 10 mg/kg, respectively. Only slight differences in percentage distributions of steryl esters depending on the geographical origin of the material were observed. The patterns were shown to remain unchanged after deodorization. The method described might be a valuable tool for authenticity assessment of cocoa butter. PMID- 11407582 TI - Solid-phase extraction with styrene-divinylbenzene sorbent for high-performance liquid or gas chromatographic determination of urinary chloro- and methylthiotriazines. AB - A solid-phase extraction (SPE) procedure on a styrene-divinylbenzene (SDB-1 cartridge) for extraction and cleaning of the triazine herbicides atrazine, simazine, ametryn, and prometryn and atrazine monodealkylated metabolites from urine samples was developed and optimised for final high-performance liquid chromatographic (HPLC-UV diode array detection) and gas chromatographic (GC electron-capture detection and GC-thermionic-sensitive detection) analyses. Interfering polar matrices were eliminated by rinsing SDB-1 with 1% acetonitrile in water or with pure water. Extraction recoveries were from 78 to 101% with an RSD of about 10% for all studied compounds. The extraction recovery for the didealkylated atrazine metabolite was significantly lower and this compound cannot be determined with these procedures. Sorbent matrix generated interferences, although not detected by the chromatographic system, lowered the response of nitrogen-phosphorus and electron-capture GC detectors for monodealkylated chlorotriazines when compared to standards prepared in n-hexane. HPLC and GC analysis with SPE (SDB-1) preconcentration showed excellent linearity over the concentration range tested, with detection limits in urine of 10 ng ml( 1) for the parent herbicides (HPLC and GC analysis) and 20 ng ml(-1) for monodealkylated chlorotriazines (HPLC analysis). PMID- 11407583 TI - Different elution modes and field programming in gravitational field-flow fractionation. III. Field programming by flow-rate gradient generated by a programmable pump. AB - Gravitational field-flow fractionation (GFFF) utilizes the Earth's gravitational field as an external force that causes the settlement of particles towards the channel accumulation wall. Hydrodynamic lift forces oppose this action by elevating particles away from the channel accumulation wall. These two counteracting forces enable modulation of the resulting force field acting on particles in GFFF. In this work, force-field programming based on modulating the magnitude of hydrodynamic lift forces was implemented via changes of flow-rate, which was accomplished by a programmable pump. Several flow-rate gradients (step gradients, linear gradients, parabolic, and combined gradients) were tested and evaluated as tools for optimization of the separation of a silica gel particle mixture. The influence of increasing amount of sample injected on the peak resolution under flow-rate gradient conditions was also investigated. This is the first time that flow-rate gradients have been implemented for programming of the resulting force field acting on particles in GFFF. PMID- 11407584 TI - Multiresidue analysis of insecticides in soil by gas chromatography with electron capture detection and confirmation by gas chromatography-mass spectrometry. AB - A rapid multiresidue method has been developed for the analysis of nine insecticides (organochlorines, pyrethroids and organophosphorus) in soil. The method is based on the sonication extraction of residues from a certain amount of soil placed in a small column, using ethyl acetate. The effect of the residence time of insecticides in soil, the material of the columns used (glass or plastic columns) and the soil moisture content on the recovery of these compounds was also studied. Residues were determined by gas chromatography with electron capture detection. The average recovery through the method obtained for these compounds varied from 90 to 108% with a relative standard deviation between 1 and 11%. The results of this study pointed out that the recoveries of insecticide residues obtained with plastic or glass columns at different soil moisture content were similar and that the residence of these compounds in soil during several days did not affect their recovery from soil. Confirmation of residue identity was performed by gas chromatography coupled with mass spectrometry. PMID- 11407585 TI - Post-capillary reaction detection in capillary electrophoresis based on the streptavidin-biotin interaction. Optimization and application to single cell analysis. AB - A class-selective post-capillary reaction detection method for capillary electrophoresis is described in which a streptavidin-fluorescein isothiocyanate (streptavidin-FITC) conjugate is used to detect biotin moieties. The selective binding of biotin moieties to the streptavidin-FITC conjugate causes an enhancement of fluorescence proportional to the concentration of biotin present. After capillary electrophoresis the separated analytes react with streptavidin FITC in a coaxial reactor and are then detected either by a benchtop spectrofluorometer (2.5 microM detection limit) or by an epi-fluorescence microscope (1 x 10(-7) M detection limit). The method is used to examine biotinylated species in a crude mammalian cell lysate which was found to contain 83+/-3 fmol in 3600 cell volumes. In addition, it is used to examine the uptake of biotin by individual sea urchin oocytes. The results indicate that, in the oocytes, biocytin is the prevalent form of biotin and its concentration varies widely between cells (mean=2+/-2 microM). PMID- 11407586 TI - Mass transport characteristics and chromatographic behavior of preparative electrochromatography on hydroxyapatite. AB - Introducing an electric field into chromatography on hydroxyapatite (HAP) was attempted in order to enhance mass transfer and separation performance. A membrane spaced multicompartment electrolyzer was developed for electrochromatography on HAP. The high performance of liquid transport by electroosmotic flux was identified and described in terms of dynamic electroosmotic pressure. The application of the electric field resulted in an improved adsorption of bovine serum albumin as shown by the breakthrough curve as function of the electric field. An improved elution was also obtained in the presence of the electric field. The results show that electroosmosis is a powerful tool of liquid transport and dispersion in a packed bed of fine particles and has potential in the large-scale chromatography of biological molecules. PMID- 11407587 TI - Characterization of phosphorylation of a novel protein kinase in rice cells by capillary electrophoresis. AB - It was demonstrated that a separation of 20 amino acids constituting a protein and three phosphoamino acids that mostly frequently occur in eukaryotes was achieved within 15 min by capillary electrophoresis coupled with lamp-induced fluorescence detection. Fluorescein isothiocyanate was employed as the fluorescence label to facilitate the fluorescence detection of the 23 amino acid species. The fluorescent derivatization conditions and separation parameters including concentration of electrolyte, surfactant in buffer, applied voltage and sample injection were investigated in detail and optimized. The influence of buffer additives such as methanol, acetone and polyvinylpyrrolidone on separation selectivity and sensitivity were discussed. We showed that addition of 2% polyvinylpyrrolidone into the running buffer could dramatically enhance the separation selectivity of amino acids at the expense of a decrease of sensitivity of phosphoamino acids. Under the optimized conditions, the detection limits (S/N=2) ranged from 1.90 x 10(-8) M to 5.66 x 10(-8) M with an average efficiency of 620,000/m. The method was applied to characterization of the phosphorylation of a novel protein kinase RCaMBP (calcium/calmodulin-binding protein kinase) encoded by a cDNA newly isolated and cloned from rice. We verified that RCaMBP belonged to a type of Ser/Thr kinase, providing insight into its function in signal transduction. PMID- 11407588 TI - Capillary zone electrophoresis for the determination of light-absorbing anions in environmental samples. AB - A rapid and simple method for separation and determination of inorganic anions by capillary zone electrophoresis was described. The detection was carried out directly with a diode array detector. The experimental conditions, such as concentration of carrier electrolyte, capillary length, voltage, and temperature were optimized. In order to improve selectivity, different organic modifiers were also investigated. The baseline separation of 10 light-absorbing anions was accomplished within 3.5 min with a background electrolyte consisting of 50 mM sodium tetraborate containing 5% MeOH. Linear plots were obtained in the concentration range of 0.1-10 microg/ml. With sample stacking injection, the quantitation limits of the anions were found to be in the range of 0.02-0.1 microg/ml. The proposed method was successfully applied to the determination of inorganic anions in environmental samples and in effluents of a power plant. PMID- 11407589 TI - Ammonia as carrier gas in capillary gas-liquid chromatography. AB - Influence of helium and ammonia as carrier gases on retention of neutral solutes on polyethylene glycol has been studied theoretically and experimentally. It was shown that the influence of ammonia on the retention varied significantly depending on the solute's nature. For example, the influence is great for C22 n alkane and it is small for phenol. Taking into account the results obtained by various researchers it makes sense to express an opinion on the expediency of development of gas chromatography operated under the conditions of acidic-basic chromatography. PMID- 11407590 TI - Pressurised fluid extraction of bupirimate and ethirimol from aged soils. AB - This paper assesses the effect of pressurised fluid extraction (PFE) on the recovery of bupirimate and its degradation product, ethirimol from a range of soil types. The analytes were extracted under standard conditions (pressure, 2000 p.s.i.; temperature, 100 degrees C; and, three static flush cycles of 5 min static extraction time each) using a variety of individual and combined solvents. It was found that the recovery of bupirimate was dependent upon the organic matter content of soil. PMID- 11407591 TI - Determination of three nitroimidazole residues in poultry meat by gas chromatography with nitrogen-phosphorus detection. AB - A method was developed for the determination of the nitroimidazole compounds dimetridazole (DMZ), ronidazole (RNZ) and metronidazole (MNZ) by gas chromatography with nitrogen phosphorus detection. Nitroimidazole compounds were extracted with acetonitrile, followed by acidification using acetic acid and cleanup using strong cation-exchange (SCX) SPE column. Validation in chicken muscle fortified at a concentration of 5 microg/kg gave mean recoveries of 85% DMZ, 90% RNZ, 80% MNZ with RSDs of 13.0, 14.3, 11.2%, respectively (n=6). The method is suitable for statutory residue testing and is used as a quick screening method in the National Residue Surveillance Plan in China. PMID- 11407592 TI - Integration of the pRB and p53 cell cycle control pathways. AB - Two fundamental molecular pathways, the pRB and p53 pathways, regulate cell growth and cell death. The importance of these pathways in cellular growth control is underscored by the observation that members of these pathways are found mutated in all human cancers. These two pathways have typically been studied and described independently. However, as we discuss here, recent data have revealed an intimate molecular and genetic interaction between the p53 and pRB pathways. PMID- 11407593 TI - Cip/Kip cell-cycle inhibitors: a neuro-oncological perspective. AB - The cell cycle is a precisely controlled cellular program that ensures normal cellular proliferation and development. The cyclin-dependant kinases (CDK) are molecules central to the continued progression through the cell-cycle checkpoints and as such are regulated by various mechanisms including cyclin levels, phosphorylation/dephosphorylation and cyclin-dependant kinase inhibitors (CKI). The CKIs are grouped into two families based on their structure and function, four lnk4 CKIs and three Cip/Kip CKIs. Abnormalities in these proteins can give rise to developmental defects and cancer. In this review, we will discuss the biochemistry and cell biology of the each of the Cip/Kip CKIs, their role in development as evidenced by targeted mutations in mice, and their role as possible tumor suppressor genes. PMID- 11407594 TI - The INK4A/ARF locus: role in cell cycle control and apoptosis and implications for glioma growth. AB - The unique INK4A/ARF locus at chromosome 9p21 encodes two distinct proteins that intimately link the pRB and p53 tumour suppressor pathways. p16INK4A has been identified as an inhibitor of the cell cycle, capable of inducing arrest in G1 phase. p14/p19ARF on the other hand can induce both G1 and G2 arrest due to its stabilizing effects on the p53 transcription factor. In addition to their roles in growth arrest, both proteins are involved in cellular senescence and apoptosis. The frequent mutation or deletion of INK4A/ARF in human tumours as well as the occurence of tumours in the murine knockout models have identified both p16 and ARF as bona fide tumour suppressors. PMID- 11407595 TI - The role of p53 in human cancer. AB - In the two decades since its original discovery, p53 has found a singularly prominent place in our understanding of human cancer. Although the biochemistry of p53 has been worked out in some detail, our knowledge of the biologic consequences of p53 dysfunction is still quite rudimentary. Over the next several years, it will be important to determine how best to harness the complex properties of p53's ability to induce cellular growth arrest and cell death to generate novel, effective approaches to cancer therapy. Furthermore, a clearer appreciation of the direct interaction of epigenetic factors with p53 will lead to development of strategies to inhibit tumour initiation and progression. The next decade promises to offer exciting opportunities to apply our vast knowledge of this intriguing tumor suppressor to clinical advantage. PMID- 11407597 TI - Animal models of cell cycle dysregulation and the pathogenesis of gliomas. AB - Mutations in gliomas, for the most part, fall into two main categories. The first category of mutations affects genes that produce proteins which activate signal transduction pathways downstream of tyrosine kinase receptors; the second category disrupts the pathways leading to cell cycle arrest. Cell cycle arrest pathways normally maintain cells in the G1 phase of the cell cycle, preventing inappropriate proliferation. The role of disregulation of these pathways in tumor formation is currently the focus of many investigations. Studies carried out with astrocytes and other cell types indicate that these pathways may also function in maintenance of appropriate chromosome number and differentiated phenotype, and in acquisition of senescence. Genetically defined mouse models of gliomagenesis have been helpful in increasing our understanding of how cell cycle arrest pathways cooperate with alterations in signal transduction pathways to provoke tumor formation in many cell types, including glial cells. Various strategies for experimental cell cycle arrest disruption show minimal or no formation of gliomas. In contrast, gliomas are generated with a number of strategies that enhance signal transduction downstream of tyrosine kinase receptors. Experimental disruption of the cell cycle arrest pathways is required for gliomagenesis in some of these models, but not in others. Furthermore in some cases, although not required for gliomagenesis, disruption of the cell cycle arrest pathways appears to enhance glioma formation. The results of these mouse model experiments imply a potentially complex role for cell cycle arrest disruption in human gliomagenesis. PMID- 11407596 TI - Mitogenic signaling and the relationship to cell cycle regulation in astrocytomas. AB - The activity and regulation of a number of mitogenic signaling pathways is aberrant in astrocytomas, and this is thought to play a crucial role in the development of these tumors. The cascade of events leading to the formation and the progression from low-grade to high-grade astrocytomas is well characterized. These events include activating mutations, amplification, and overexpression of various growth factor receptors (e.g. epidermal growth factor receptor (EGFR), platelet derived growth factor receptor (PDGFR), c-Met), signaling intermediates (e.g. Ras and Protein kinase C (PKC)), and cell cycle regulatory molecules (e.g. mouse double minute-2 (Mdm2), cyclin-dependent kinase-4 (CDK4), and CDK6), that positively regulate proliferation and cell cycle progression. Inactivating mutations and deletions of signaling and cell cycle regulatory molecules that negatively regulate proliferation and cell cycle progression (e.g. p53, p16/INK4a, p14/ARF, p15/INK4b, retinoblastoma protein (Rb), and Phosphatase and tensin homologue deleted from chromosome 10 (PTEN)) also participate actively in the development of the transformed phenotype. Several mitogenic pathways are also stimulated via an autocrine loop, with astrocytoma cells expressing both the receptors and the respective cognate ligand. Due to the multitude of factors involved in astrocytoma pathogenesis, attempts to target a single pathway have not given satisfactory results. The simultaneous targeting of several pathways or the targeting of signaling intermediates, such as Ras or PKC, situated downstream of many growth factor receptor signaling pathways may show more efficacy in astrocytoma therapy. We will give an overview of how the combination of these aberrations drive astrocytoma cells into a relentless proliferation and how these signaling molecules may constitute relevant therapeutic targets. PMID- 11407599 TI - Orientational information from TEDOR spectral sidebands. AB - In a dipolar-coupled spin-1/2 network of the type 15N1-(13)C-15N2, an assessment of the sensitivity of the N --> C and C --> N TEDOR sideband intensities to the Euler angles defining the orientation of the two heteronuclear dipolar vectors in the 13C and 15N chemical shift (CS) tensor principal axes system has been carried out via numerical calculations. The results clearly indicate the potential of TEDOR MAS NMR spectroscopy for the characterization of the CS tensor orientation in the molecular frame. The efficacy of the method has been experimentally illustrated by TEDOR studies on a polycrystalline sample of [1, 3-(15)N2, 2 (13)C]uracil, which is one of the four bases in RNA. PMID- 11407598 TI - Delivery of cell cycle genes to block astrocytoma growth. AB - Current therapies for glioblastoma multiforme are ineffective. Therefore, novel therapies that target specific differences between normal and malignant cells are urgently needed. Abnormalities of cell-cycle related genes are a common feature of cancer in general and astrocytic tumors in particular. The role of these proteins is to help to regulate cell proliferation, differentiation and apoptosis. Restoring wild-type activity of critical regulators of the cell cycle to astrocytic tumors generally results in modification of the growth properties, and often the viability, of the cancer cells. Transfer of p53 induces growth arrest and, more importantly, apoptosis. Restoration of the Rb pathway results in either reversible growth arrest or senescence. Expression of E2F-1 induces transient increase of proliferation followed by massive apoptosis. Overexpression of MMAC/PTEN arrests cell cycle progression in G1 and promotes anoikis. Current knowledge of the functions of these cell-cycle controllers can be used to design small peptides and drugs to induce cell-cycle related anti-cancer effect. Inactivation of the p53 and Rb pathways in cancer cells is also being used to engineer mutant viruses that are able to replicate exclusively in cancer cells. PMID- 11407600 TI - Ammonium tunnel levels and spin-rotational wavefunctions in (NH4)2S2O8. AB - Proton spin-lattice relaxation in a single crystal of (NH4)2S2O8 was studied as a function of resonance frequency at various constant temperatures between 4.2 and 30 K. Two T1 minima were found, one at 8.6 MHz and the other at 4.3 MHz. They are related to the splitting between the lowest T level and the A level, equal to 8.6 MHz nearly independently of temperature below 25 K. Together with the large tunnel splitting of 269 MHz, determined earlier by Clough et al. (Chem. Phys. 152, 343 (1991)) our results define the spin-rotational wavefunction of the lowest T level very accurately, although those of the two higher T levels remain largely undetermined. PMID- 11407601 TI - Quantitative structure-property relationships in boron nitrides: the 15N- and 11B chemical shifts. AB - Nuclear Magnetic Resonance (NMR) chemical shifts(delta) for elements in solids may often be approached by ab initio cluster calculations. We employ this technique to investigate the influence of structural alterations on the 15N and 11B chemical shifts in boron nitrides--in both hexagonal and cubic modifications. Within a given class of connectivity, i.e., three- or fourfold coordinated nitrogen, for the first time, an almost linear correlation between the 15N chemical shift and N-B bond lengths could be established. Also, the 11B shifts in hexagonal boron nitride correlate with the B-N bond distance; however, the effect is less pronounced. For the value of the chemical shift (CS), the decisive property is the average bond length at the atom in focus. Variations of CS are predominantly caused by changes in the paramagnetic deshielding. Further, second nearest neighbor effects on the shieldings at 15N nuclei are quantified by subtraction schemes. The present work is closely related to the verification of models for amorphous high-demand Si/B/N ceramics. PMID- 11407602 TI - Niobium-93 MQMAS NMR spectroscopic study of alkali and lead niobates. AB - 93Nb (I = 9/2) multiple-quantum magic-angle spinning (MQMAS) NMR spectra of a series of inorganic niobates have been measured. 93Nb MQMAS spectroscopy yields spectra with typically an order of magnitude higher resolution than that obtainable with 93Nb MAS spectroscopy and 93Nb dynamic-angle spinning (DAS) spectroscopy. For example, the full-width at half-maximums of the 93Nb resonances of LiNbO3 were 9 (MAS), 5.8 (DAS), and 0.7 kHz (MQMAS). Broadening of the 93Nb MAS and DAS spectra is due to the second-order quadrupolar and homonuclear dipolar interactions, respectively. The quadrupolar products (P(O)) and isotropic chemical shifts (delta(iso)) of the 93Nb resonances of LiNbO3, NaNbO3, PbNb2O6, Pb2Nb2O7, Pb3Nb2O8, Pb3Nb4O15, Pb3Nb4O13, and Pb1.83Nb1.71Mg0.29O6.39 were determined from MQMAS spectra and range from 13.6 to 26.8 MHz and from -951 to 1113 ppm, respectively. Resonances with relatively large quadrupolar coupling constants (> 30 MHz) were not observed using MQMAS spectroscopy, but were detected using nutation spectroscopy. The applicability and limitations of MQMAS spectroscopy in studying inorganic niobates containing multiple 93Nb resonances are addressed and compared with those of MAS, nutation, and DAS spectroscopies. PMID- 11407603 TI - A movement for the protection of infancy: what we must do to lower child mortality in Papua New Guinea. PMID- 11407604 TI - Anorectoplasty in children in Papua New Guinea. AB - The posterior sagittal anorectoplasty (PSARP) procedure for the definitive repair of children with imperforate anus was described in 1982. Unfortunately, surgeons in Papua New Guinea (PNG) have until recently not had the opportunity of being trained in the technique. Through the Medical Officer, Nursing and Allied Health Sciences Training Project (MONAHP) and Pacific Islands Project (PIP) of the Royal Australasian College of Surgeons, 65 Papua New Guinean children with an anorectal anomaly have undergone a repair, in conjunction with training of the surgical staff and medical students. A new technique for the management of a prolapsed colostomy has been developed and a protocol for management of PSARP patients postoperatively has been formulated. Patients referred to the paediatric surgical visiting teams were diagnosed and treated according to the stage their management had reached. Patients with a low anomaly were treated by a cutback procedure, those with a colostomy and a high lesion were managed by a PSARP and those with failed previous surgery were managed with a redo anorectoplasty, often without a covering colostomy. Data were collected on the patients treated and, where possible, the patients were followed during subsequent visits. 65 patients with an anorectal anomaly were treated, of whom 6 were treated with a cutback and 43 had a primary repair of a major anomaly. 5 of these 43 involved an abdominoperineal procedure. 19 children had redo surgery, 3 of whom had a second operation by the senior author, due to failure of initial postoperative management; 1 of these was for a failure to carry out the postoperative dilatations and 2 were due to poorly controlled constipation in the early postoperative period. A protocol for the postoperative dilatations was developed using shaped candles. Major complications were uncommon, in particular infections were rare despite the relative lack of facilities. However, difficulties with outpatient follow-up resulted in problems that could have been avoided. A large number of anorectal anomalies have been successfully treated as part of the MONAHP and PIP projects with local surgeons learning the technique. A protocol for follow-up and a technique for the management of colostomy prolapse have been developed. PMID- 11407605 TI - Perinatal asphyxia at Port Moresby General Hospital: a study of incidence, risk factors and outcome. AB - We investigated the incidence and outcome of perinatal asphyxia (PA) at Port Moresby General Hospital by a retrospective chart review and prospective collection of data, spanning a total of 2.5 years. 125 babies weighing more than 2000 g at birth with a gestation of 34 weeks or more and with no obvious congenital abnormalities were diagnosed to have PA. During the same time period 22,700 liveborn babies were delivered, a PA incidence of 5.5/1000 livebirths. There was a 31% mortality and considerable morbidity. Hospital records for 114 affected babies and 115 controls (the next baby born by normal delivery) were compared. Significant risk factors for PA were: previous stillbirth or neonatal death, fetal heart rate abnormalities, membranes ruptured for more than 12 hours prior to delivery, meconium staining, antepartum haemorrhage, maternal fever, prolonged first and second stages of labour, preterm or post-term delivery and operative delivery. In only 73 affected babies was the 5-minute Apgar score recorded as 6 or less. All 34 of the babies with grade 3 hypoxic ischaemic encephalopathy (HIE) either died (30) or had serious neurological impairment. The treatment of affected babies remains largely supportive and some causes of PA are currently unavoidable. It is, however, widely accepted that some cases of perinatal asphyxia may be prevented by the delivery of high-risk pregnancies in obstetric facilities with appropriate intervention and by good neonatal resuscitation. Sophisticated or expensive equipment is not a necessity. PMID- 11407606 TI - Congenital syphilis at Goroka Base Hospital: incidence, clinical features and risk factors for mortality. AB - A prospective study of all cases of congenitally acquired syphilis diagnosed at Goroka Base Hospital was conducted between January 1998 and December 1999. 67 affected neonates and children were seen, of whom 19 died during the first admission and 3 died during the period of follow-up. Congenital syphilis caused 5.5% of 994 neonatal admissions, but 22% of all neonatal deaths. The major risk factor for death in affected babies was low birthweight. A birthweight of less than 2 kg had an odds ratio for death of 30.0 (95% confidence interval 6.90 131.1). During the time of the study 5385 women attended antenatal care at Goroka Base Hospital, of whom 382 had both positive VDRL and TPHA tests. The incidence of syphilis in women attending antenatal care was 7.1%. Syphilis is a major cause of neonatal mortality and morbidity and a major cause of morbidity among women of childbearing age in the Eastern Highlands Province. Current antenatal screening is inadequate, covering less than 30% of pregnant women. New ways to extend syphilis screening and treatment to all affected pregnant women are urgently required and must be formally assessed. PMID- 11407607 TI - The effect of introduction of minimal standards of neonatal care on in-hospital mortality. AB - A retrospective study was done to assess the effect on in-hospital neonatal mortality of a series of interventions in neonatal care in the highlands of Papua New Guinea. Between 1995 and 1997, prior to the interventions, the mortality among neonates admitted to the Goroka Hospital Special Care Nursery was 18% and two-thirds of very low birthweight (1-1.5 kg) neonates died. The interventions began in December 1997 and were aimed at reducing mortality among all neonates and particularly among those with very low birthweight. Compared to the 30-month period prior to the interventions, the in-hospital neonatal mortality in the 30 month period after the interventions began was 44% lower (relative risk (RR) 0.56, 95% confidence interval (CI) 0.45-0.69). After adjustment for a higher number of neonates <1500 g in the pre-intervention period, the relative risk was 0.59 (0.48-0.74). The mortality in the intervention phase for very low birthweight babies was 56% lower (RR 0.44, 95% CI 0.30-0.65) and for moderate low birthweight (1.5-2 kg) 50% lower (RR 0.50, 95% CI 0.28-0.90). Mortality was also significantly lower in the intervention phase in neonates with a diagnosis of septicaemia or pneumonia (RR 0.36, 95% CI 0.19-0.67), but there were no differences in mortality from birth asphyxia, meconium aspiration or extremely low birthweight (<1 kg). We estimate that in the 30 months after beginning the interventions 82 neonatal deaths that would previously have occurred were avoided. The costs of the improvements in technology described are estimated at US$445 (K1000) per life saved, but substantial training and improved supervision of staff and other human factors may have been more important than equipment. Apnoea monitors were the single most important technology introduced. A similar evaluation of the effect of minimal standards should be done without the use of incubators and overhead heaters, as these are costly and may be dangerous when used by less experienced operators. The 33 neonatal deaths that we estimate were avoided each year because of the interventions represent less than 10% of all neonatal deaths in the province. Although this study provides justification for increasing the technology for supportive neonatal care and training in medium sized hospitals in rural areas in developing countries, estimates of cost effectiveness must be compared with other interventions that will effectively lower neonatal mortality, both in and out of hospitals. PMID- 11407608 TI - Current opportunities and future challenges in child health in Papua New Guinea. AB - Papua New Guinea (PNG) is at a crossroads in child health care. We know how to treat or prevent most of the diseases causing child and maternal mortality, but we are failing to deliver the services to where most of the people are dying. There is much to be learnt from early achievements in the development of health services in PNG. Between the 1950s and 1970s the infant mortality rate fell from 250-500 per 1000 livebirths to 77-100 per 1000 livebirths. This occurred predominantly because of an emphasis on public health programs and cost-effective standard treatment. Now the public health services have deteriorated, vaccine coverage is poor and health workers are not fully using the standard treatment resources available to them. The greatest academic and practical challenges facing medicine in PNG are to implement effective and consistent delivery of vaccines, and nutritional, basic curative and maternal care to remote or impoverished communities. The other unmet prerequisites for improvement in health status are that communities participate in and take some responsibility for their health care needs. There are now unprecedented opportunities for collaboration with international partners, churches, non-government organizations and community groups to achieve these goals. Specific key strategies in child health include the more widespread utilization of the 10-Step Checklist, adoption of some of the World Health Organization Integrated Management of Childhood Illness strategies, increased output of child health nursing training, improving vaccine delivery and cold-chain services, and new immunization strategies for acute respiratory infections. The National Health Plan 2001-2010, launched in August 2000, emphasizes the vital importance of public health; the need for greater access to quality health services; improving clinical skills among health workers at all levels; and cooperation between all interested parties. By 2010 the National Health Plan must be judged not on how good the plan seemed on paper, but on whether there was a measurable reduction in maternal and child mortality over the first decade of this century. PMID- 11407609 TI - The grass is growing up around all the houses. PMID- 11407610 TI - Minimal standards of neonatal care in hospitals. PMID- 11407612 TI - The implementation of the United Nations Convention on the Rights of the Child: global commitments and local realities. AB - Papua New Guinea (PNG) is a signatory to the United Nations Convention on the Rights of the Child (CRC) and the World Health Organization Alma Ata Declaration of Health for All by 2000. The inherent rights of all children include accessible, affordable and quality health care and protection from all forms of violence, abuse, neglect and maltreatment. In 2000 PNG is challenged in implementing the CRC and has deferred achievement of targets in the Alma Ata declaration until 2010. Constraints to CRC implementation include lack of public awareness about the Convention, even among people whose work directly relates to child issues; deterioration in many essential social services; massive social change; and decentralization of health care systems without adequate accompanying support. New health problems, including alcohol and drug abuse and AIDS, now face many children and adolescents, but the problems that have existed for generations, particularly acute respiratory infections, tuberculosis and malnutrition, have not yet been dealt with. It is unlikely that primary health services can be rebuilt as they were 20 years ago and for most children throughout PNG the basic right of access to quality health care is not a reality. Programs for training village-based child health and child protection workers, who are volunteers and who are integrated into the loose health network that currently exists, would fill some of the gaps. More recognition and funding are needed from governments for church and non-government organizations involved in successful existing maternal and child health programs. There needs to be a substantial public awareness program on child rights, establishment of a mandated national coordinating body and a realistic timetable for CRC implementation. Both political will and administrative action are desperately needed. PMID- 11407613 TI - Immunization--dramatic new evidence. AB - The current EPI (Expanded Programme on Immunization) vaccines do not specifically target the organisms that lead to the two main causes of death in children - pneumonia and diarrhoea. This implies that the EPI vaccines will have only a modest effect on total child mortality. However, recent evidence suggests that measles and BCG vaccines dramatically reduce child mortality through nonspecific effects - that is, they reduce mortality from many causes, not just measles and tuberculosis. The combination of BCG at birth and measles vaccine at 6 months probably reduces total mortality to about one-third of its previous level. This means that immunization must now have the very highest priority. If we could improve immunization in Papua New Guinea so that all children received BCG, measles, diphtheria-pertussis-tetanus and polio vaccines, we would reduce child mortality from 120 to approximately 52 per 1000 livebirths - a truly spectacular reduction. The old polysaccharide pneumococcal vaccine is safe and effective and bulk purchases are likely to cost US$1 a dose or less. Further studies are needed of the effects of pneumococcal vaccine. Immunization of mothers and babies might reduce child mortality by 20%, at a cost of only US$83 per life saved. The available evidence suggests that one dose of pneumococcal vaccine given to every Papua New Guinean over 5 years of age every 5 years would save approximately 6600 lives a year and the vaccine would cost only US$121 per life saved. It will not be easy to achieve high immunization rates throughout Papua New Guinea. Vaccines will have to be given the highest possible priority, with curative medical services secondary to immunization. Health workers, government, the general population and overseas donors will have to be convinced of the very great benefits that will come from effective immunization. A sustained education campaign will be needed in addition to the establishment of an effective delivery system. The time has come for a radical shift in emphasis in Papua New Guinea: from hospitalization to immunization. PMID- 11407614 TI - Urbanization and child health. PMID- 11407615 TI - On reducing childhood trauma: child accident prevention--the poor relation of preventive health. A tribute to the late Professor John Biddulph OBE. PMID- 11407616 TI - Detection and treatment of childhood malnutrition in Papua New Guinea. AB - This paper reviews the importance of childhood malnutrition in Papua New Guinea and other developing countries. Emphasis is given to two important causes: chronic disease and limited food supply. Definitions and diagnostic criteria for malnutrition are discussed with particular focus on the mid-upper-arm circumference (MUAC). The design of a MUAC measuring tape with written treatment guidelines for use at the aid post level and a pilot project used to test the tape are described. The rationale for six treatment steps for malnutrition are described. The importance of community education, support through church groups and effective ongoing preventive health programs at the village level are stressed. PMID- 11407617 TI - Tributes to and remembrances of Professor Robert Arthur John Biddulph, OBE, MB BS, DTM&H, DCH, MRCP (Edin), FRCP (Edin), FRACP 15 January 1935 to 6 January 1998. PMID- 11407618 TI - Medical postgraduate education in child health in Papua New Guinea. PMID- 11407619 TI - The development of paediatric surgery in Papua New Guinea. AB - Until 1993 complex surgery for children in Papua New Guinea (PNG) was usually conducted by general surgeons, or by overseas paediatric surgeons during ad hoc visits. There had been little coordination of these occasional international visits and no involvement in the surgical teaching program in the University Department of Surgery. Over eight years from July 1993 to July 2000, three Australian surgeons spent 33 weeks teaching paediatric surgery and paediatric urology, using lectures, tutorials and operative demonstration sessions. This paper is a report of the teaching and service provided by one surgeon (PAD) over 27 weeks in PNG and describes the development and training of the surgeon who will be the country's first qualified paediatric surgeon (MM). Other support given has been assistance with the publication of scientific papers, reviews of Masters' theses, the development of a proposal for investigation of the prevalence of renal tract anomalies and the development of protocols for the surgical management of anorectal anomalies and Hirschsprung's disease. In addition to this teaching, 311 children have had surgery during the 11 visits that form the basis of this report. The visits have been supported and funded by AusAID and the Royal Australasian College of Surgeons through the Medical Officer, Nursing and Allied Health Professional (MONAHP) and Pacific Islands Project (PIP) programs. An indication of the impact on the care of children with surgical diseases is evident from the improved skills and the changed referral patterns over the eight years. PMID- 11407620 TI - A history of surgery for congenital heart disease in Papua New Guinea. AB - Cardiothoracic surgery in Papua New Guinea (PNG) was somewhat ad hoc prior to 1956 but later settled into an arrangement in which visiting teams from overseas selected mostly adult patients for a limited range of closed heart operations to be done locally or overseas. In 1978 the late Professor John Biddulph was instrumental in facilitating a more formal arrangement with the Royal Prince Alfred Hospital whereby patients were selected by a visiting cardiologist on an annual basis to be transferred to Sydney for cardiac surgery. This subsequently developed into a predominantly paediatric program based at the Royal Alexandra Hospital for Children in Sydney, which successfully ran until 1992. In 1993 a program began, based at the Sydney Adventist Hospital, in which a voluntary cardiac team has been visiting annually to perform both open and closed heart surgery. This program has proved to be very successful with a high output and a low mortality. Despite this long history of surgery for congenital heart disease in PNG, no definite long-term plans have eventuated. Because the financial situation of the country does not allow for a major cardiothoracic unit, the current arrangement whereby noninvasive investigation and some closed surgery are performed at Port Moresby General Hospital is appropriate for the foreseeable future. PMID- 11407621 TI - The use of the Paediatric Standard Treatment Book by clinic and health centre staff. AB - The study assessed the self-reported frequency and quality of use of the Paediatric Standard Treatment Book by staff in urban clinics and rural health centres. 61 of the 88 nursing officers and 44 of the 89 community health workers in 9 urban and 4 rural health settings completed written questionnaires on their use of the Standard Treatment Book. The survey participants were also assessed on the management of three case scenarios of common clinical conditions. Whilst 69% of the participants reported daily use of the book, only 51% indicated that they always followed the guidelines. Performance in the case scenarios was poor. Although 87% made a correct diagnosis in the most straightforward case, only 38% indicated complete treatment and only 36% indicated complete and correct advice. In two more complex scenarios less than 30% of the participants made correct diagnoses and less than 10% indicated complete treatment and advice. 75% of the study group wanted inservice training on the use of the book; the majority of these said that doctors should give this training. 79% thought that the book could be improved. Many of the participants felt that more topics and more flow charts should be included. Whilst nursing officers and community health workers regard the Standard Treatment Book (STB) as important, many do not make optimal use of it. Knowledge of appropriate advice to give parents regarding their child's illness was particularly poor. Given the low scores of health workers on case scenarios involving children with more than one presenting problem, the use of the STB appears to be essential for management of most severely ill children presenting to health facilities in Papua New Guinea. Doctors, especially paediatricians, have an important role to play in stressing the importance of the book, in teaching health workers to use it correctly and in emphasizing an integrated approach to the management of sick children. The study incorporated an assessment of health facility infrastructure and equipment. All facilities needed maintenance work, and more than half had significant deficiencies in equipment and drug availability. Medical staff supervision and support of primary health staff is important and should include increasing and improving the use of the Standard Treatment Book. Such support should also aim to improve the working environment and health facility resources. This would substantially improve the service provided. PMID- 11407622 TI - The role of non-government organizations in supporting and integrating interventions to improve child health. AB - In Papua New Guinea there are many organizations providing sparsely spread and fragmented health services. Government health facilities are often relatively well functioning in urban and periurban areas, but sporadic or nonexistent in rural areas. In some remote areas churches are the major health service providers. Increasingly other community groups are providing village-based health services. Much financial support is now pledged by major international donors for community-based health services, but few people working at a district or community level have the management skills to access the funds or plan programs effectively, and few of the major donors have any significant presence in rural areas. Such a management skill gap also exists at the level of many provincial health offices and this seriously limits the effectiveness of all major donor projects. There is need for integration of health services to avoid replication and to extend services to areas where no effective services are currently provided. There is also a great need to train people at a community and district level in program planning and management. Non-government organizations (NGOs) working at a district or community level have the potential to bridge this skill gap and to help integrate community-based services with government institutions. This paper reports, as an example, the activities of Save the Children, an international NGO in Papua New Guinea. Essential for the success of community based health projects is the development of local management skills, reliable funding, integration with established health institutions, objective evaluation and community support. Skilled NGOs working at a community, district or provincial level can have important roles in assisting local people to run effective and sustainable health programs. PMID- 11407623 TI - Antibiotic-resistant bacterial sepsis in Papua New Guinea. AB - Infections due to antibiotic-resistant bacteria, especially gram-negative bacteria, are a common cause of child mortality in Papua New Guinea. Antibiotic resistant bacteria include the enteric gram-negative bacilli, especially Escherichia coli, Klebsiella and Enterobacter, and Haemophilus influenzae type b, a major respiratory tract pathogen and cause of meningitis. Among these bacteria there is now high-level resistance to standard antibiotics, including chloramphenicol, amoxycillin and cotrimoxazole. Reasons behind the increase in antibiotic-resistant bacterial infections are the widespread unregulated use of antibiotics and the very large burden of bacterial infections. Risk factors for development of resistant enteric gram-negative infections include village births, prolonged hospital stay, kwashiorkor in adopted children and previous treatment with broad-spectrum antibiotics. Cost-effective strategies to combat these pathogens will need to be broad and must focus on reducing the use of antibiotics for trivial illnesses, reducing the need to use antibiotics and reducing the risk factors for resistant bacterial sepsis. There must be stricter regulation of commercial pharmacies, education of health workers on how to avoid inappropriate antibiotic prescribing, a focus on the prevention of pneumonia by immunization with new vaccines, improvements in the quality and uptake of formal maternal care services and public health measures within villages. In addition there is a need for better surveillance for antibiotic-resistant bacteria within hospitals; this will require substantial improvements in laboratory facilities and carefully planned research collaboration. A national committee should be established to advise on these matters and coordinate interventions. PMID- 11407624 TI - Control measures and the outcome of the measles epidemic of 1999 in the Eastern Highlands Province. AB - In the Eastern Highlands Province (EHP) of Papua New Guinea (PNG) measles outbreaks have occurred regularly every 3 to 4 years since 1980. The latest was between September 1998 and March 2000. Between July 1999 and March 2000 314 children with measles were reviewed at Goroka Base Hospital. The majority of these children were very young: 55% were under 1 year and 27% under 6 months. The median age of the measles cases was 11 months (range 10 days to 13 years). 40% of the children had a verifiable history of having received at least one dose of measles vaccine. The majority were vaccinated during the epidemic and included many children who either were below 6 months of age or who developed measles within 2 weeks of vaccination. Measles complications occurred in 82% of the children, the most common being pneumonia. Serious complications, particularly severe pneumonia, were more common among the unvaccinated children than in those who had received at least a single dose of the measles vaccine. No deaths occurred among 82 children who had received measles vaccine more than 2 weeks before the onset of clinical measles, compared with 10 deaths in 206 children who had never been vaccinated against measles or were vaccinated in the 2 weeks before presentation (p=0.067). The overall case fatality was 4%: 14% among the hospital-acquired and 2.5% in community-acquired measles. Improvement in the measles vaccination coverage and supplementary vaccination campaigns are required to prevent measles outbreaks in PNG. Intensified measles vaccination campaigns, such as the one conducted in EHP in 1999, are recommended during epidemics to minimize deaths due to measles and to rapidly control outbreaks. The efficacy of measles vaccination can only be measured in total mortality, not in the prevention of clinical measles. PMID- 11407625 TI - Child adoption in the Western Highlands Province of Papua New Guinea. AB - This study investigated the epidemiology of child adoption in the Western Highlands Province of Papua New Guinea. A prospective case-control study of 100 adopted and 100 control children matched by age and sex was done in 1995. The age at the time of adoption ranged from 7 days to 8 years with 64 being adopted in the neonatal period. 28 were adopted because the biological mother had died, 23 because the adoptive mothers had been unable to bear children and 16 because the biological mother was unmarried or 'too young'. Only 11 adopted children were not blood relatives of the adoptive mother; 10 children had been abandoned and 1 had been bought for cash. 97 adoptive mothers were married. The majority (61%) had no formal education and 95% were not in paid employment. Compared with the mothers of the control children fewer adoptive mothers had received any formal education and more of them smoked cigarettes, drank alcohol or chewed betelnut. Social characteristics of the adoptive fathers were similar to the fathers of the control children. Of the 66 living biological mothers for whom information was available, 39 (59%) were married, 16 (24%) single, 8 (12%) divorced and 3 (5%) widowed. For 21 (32%) of the biological mothers the adopted baby was their first. 19 adopted babies were breastfed, 8 exclusively, 6 with the addition of non-human milk and 5 with additional solid feeds. Two-thirds of the adopted children and only 5 controls were bottle-fed. There were no significant differences in nutritional status between the two groups and immunization status was similar. There was widespread ignorance about legal adoption procedures. Only 8 adoptive mothers had any knowledge of and only 2 had followed formal adoption procedures. In this group of adopted children it appeared that most were well cared for, as their nutritional status and immunization status were similar to non-adopted children. There have, however, been suggestions that adoption is a risk factor for child abuse in Papua New Guinea and adoption has recently been associated with severe malnutrition and mortality in a highlands hospital inpatient population. Information relating to formal adoption processes should be more widely disseminated throughout Papua New Guinean communities to protect the rights of adopted children and their adoptive parents. PMID- 11407626 TI - Negotiation training and interpersonal development: an exploratory study of early adolescents in Argentina. AB - This paper reports findings from an exploratory outcome study of the Program for Young Negotiators training model with early adolescents in Buenos Aires, Argentina. Youth between the ages of 10 and 15 years (135 females, 70 males) were assessed before and after negotiation training, based on two measures of psychosocial attitudes and behavior. On the Five Factor Negotiation Scale (Nakkula & Nikitopoulos, 1999a), an increase in overall negotiation attitudes and behavior was found, with particularly large increases in the domains of conflict based perspective taking and behavioral approaches to conflict resolution. On the Relationship Questionnaire, Schultz and Selman's (1999) structural developmental measure of psychosocial competence, stronger than expected changes were found in overall competence, with fairly equal changes in the primary domains of interpersonal understanding, interpersonal skills, and the personal meaning of relationships. Finally, students who presented a pretest thought-action gap marked by a high level of interpersonal understanding relative to their level of interpersonal skill increased substantially more in negotiation attitudes and behavior than did students manifesting a gap in the opposite direction. Implications regarding who benefits most from negotiation training are discussed. PMID- 11407627 TI - Exercise is positively related to adolescents' relationships and academics. AB - Eighty-nine high school seniors were administered a questionnaire that gathered information on their exercise habits (ranging from rarely to daily), relationships with parents and peers, depressive tendencies, sports involvement, drug use, and academic performance. Students with a high level of exercise had better relationships with their parents (including greater intimacy and more frequent touching), were less depressed, spent more time involved in sports, used drugs less frequently, and had higher grade point averages than did students with a low level of exercise. PMID- 11407628 TI - Intrinsic motivation among regular, special, and alternative education high school students. AB - This study examined motivationally related variables among three types of high school students. In particular, students' perceptions of competence, control, parental autonomy support, teacher autonomy support, peer autonomy support, and academic coping were investigated. Two hundred fifty-one juniors and seniors (104 regular education, 93 alternative education, and 54 special education) from a large Southern California school district participated. Significant group differences were found on measures of perceived competence, academic coping, and parental autonomy support. Specifically, regular education students had a higher level of self-reported academic competence than did special education students. Further, regular education and special education students reported that their parents were more involved in their lives as compared with alternative education students. Finally, regular education students reported a higher level of academic anxiety than did special education and alternative education students; however, regular education students had the highest level of positive coping. The implications of these findings are discussed. PMID- 11407629 TI - Peer isolation and drug use among white non-Hispanic and Mexican American adolescents. AB - The social-emotional characteristics and drug-use patterns of adolescents who reported having no friends (i.e., isolates) were compared to those of adolescents in drug-using and non-drug-using peer groups. Adolescents who did not have drug using peers reported the lowest drug use and those with drug-using peers had the highest drug use, with adolescents who were isolated falling in between. Isolated youth reported more shyness, greater feelings of alienation, and lower social acceptance than did those in the other groups. Isolated youth also reported more anger and depression than did youth with non-drug-using peers, but less anger and equivalent depression when compared to adolescents with drug-using peers. Results are discussed in terms of social-emotional characteristics of isolated youth and risk/protective factors. PMID- 11407630 TI - Late adolescents' perspectives on marital rape: the impact of gender and fraternity/sorority membership. AB - Although recent studies of marital rape have examined both victims' and perpetrators' social and psychological characteristics, little attention has been directed to the attitudes of others toward marital rape. Using a systematic sample of college students, this study examined attitudes toward marital rape--in particular, the impact of gender and fraternity/sorority membership on respondents' (1) views regarding marital rape compared to rape by a stranger; (2) feelings about possible actions a woman who is a victim of marital rape can take; and (3) attitudes toward legislation pertaining to marital rape. It was found that college women were significantly more likely than college men to say they strongly agree that marital rape and stranger rape should be treated as similar crimes. In addition, nonfraternity men were significantly more likely than fraternity men to indicate that they strongly approve of marital rape legislation and that husbands who perpetrate marital rape should be prosecuted. Sorority membership had little impact on women's responses. PMID- 11407631 TI - A neural network approach to identifying adolescent adjustment. AB - This study examined the relationship between the quality of adjustment in adolescents and a set of psychiatric diagnoses, personality traits, parental bonding, and social support variables. One hundred fifty adolescents were administered the Millon Adolescent Personality Inventory, the Parental Bonding Questionnaire, the Social Support Questionnaire, and the Diagnostic Interview for Children and Adolescents. A neural network approach was then utilized, and it was found that several of the variables (e.g., Major Depressive Disorder, Conduct Disorder, and Societal Conformity) had a significant role in classifying adolescents into three groups: maladjusted, nominally adjusted, and well adjusted. PMID- 11407632 TI - Assessment of mode of anger expression in adolescent psychiatric inpatients. AB - Anger can play an important role in depression and suicide risk among adolescents. The present study evaluated internalized and externalized anger in 92 adolescent psychiatric inpatients. Results indicated that adolescents who internalized their anger were more likely to be depressed and to experience feelings of hopelessness. In addition, adolescents who internalized their anger made more serious suicide attempts than did those who externalized their anger. In contrast, adolescents who externalized their anger were more likely to have alcohol-related problems. Thus, different modes of anger expression appear to be related to different manifestations of psychopathology. It was concluded that assessment of mode of anger expression in adolescents may enhance our understanding of suicide and its risk factors. PMID- 11407633 TI - Identifying young people's guidance needs through telephone counseling. AB - This study examined the needs that young people in Guadalajara, in the state of Jalisco, Mexico, expressed in phone calls to a research unit of the Mexican Social Security Institute. Three hundred forty-five calls, from individuals 11 to 24 years old who phoned from June 1995 to November 1998, were analyzed. Two hundred ninety-four of the callers were female and 51 were male. Greater percentages of females than males called to ask about sexuality and family problems. Males more than females asked about reproductive health (particularly pregnancy) and mental health. There were significant differences by gender and age, differences that were not detected in health service statistics. The findings have implications for decision makers and health and education service providers. They point to the need for programs that will reinforce young people's good health practices and help them avoid risky behaviors. PMID- 11407634 TI - Integrating a structured ethical reflection curriculum into high school community service experiences: impact on students' sociomoral development. AB - It is widely accepted within the field of community service that reflection is an important factor in promoting personal and sociomoral development. The purpose of this research was to determine if a particular form of reflection--decision making with an emphasis on the ethical nature of community service--has special value in achieving service-learning goals. Three dimensions of adolescent identity development served as dependent variables: agency, social relatedness, and moral-political awareness. Four hundred seventy-six high school students were assessed on the dependent variables before and after experiencing one of three conditions: community service with an ethical reasoning component; community service with reflection, but without an ethical reasoning component; and no community service (control). After one semester, it was found that students in the first condition (ethical decision making curriculum integrated into the community service program) made greater advances on all three dimensions of identity formation when compared with students in the other two conditions. Specifically, it was found that they became more systematic in their ethical reasoning and more likely to consider situations and issues from an ethical point of view. PMID- 11407635 TI - Gender differences in psychological well-being of Mexican early adolescents. AB - This study examined gender differences in the effects of menarche in females and voice change in males, specifically with regard to depression, self-esteem, body image, and externalizing problems (i.e., behavioral disturbances). In addition, possible modifying variables (relationship with parents, social-emotional adjustment, level of parental education, and menstrual attitudes) were assessed. Participants were 1,102 Mexican youths aged 9 to 14. Analyses indicated that there were no gender differences among prepubertal youths in depression, body image, or self-esteem, but prepubertal males had more externalizing problems than did premenarcheal females. Females increased in depression, externalizing problems, and negative body image postmenarche, while males showed no change in depression, a trend toward fewer externalizing problems, and felt better about their bodies following voice change. Relationship with parents, social-emotional adjustment, parental education, and menstrual attitudes did not modify the relation between menarche and body image or depression. Based on these findings, suggestions for designing interventions aimed at early adolescent females are presented. PMID- 11407636 TI - Development and administration of a measure to assess adolescents' participation in leisure activities. AB - This paper describes the development and administration of a measure (a self report questionnaire) to assess the participation of adolescents in leisure activities. Questionnaire items were generated through a content analysis of focus group interviews with 130 young people aged 12 to 18 years. Specifically, the activities of approximately a thousand adolescents were investigated through a three-factor leisure typology--achievement, social, and time-out leisure- together with the leisure parameters of enjoyment, freedom of choice, and frequency of participation. Further, leisure engagement was examined based on gender and age (i.e., changes in activity participation across the adolescent years). The results are discussed in terms of the practical and theoretical implications of the relationship between gender and age in leisure activity participation. PMID- 11407637 TI - Examination of adolescents' negative thoughts, depressive mood, and family environment. AB - The focus of this study was the relationship among adolescents' negative thoughts, depressive mood, and family environment. Three hundred eleven students (aged 16 and 17 years) in public and private schools served as subjects. They were administered the Family Environment Scale, the Automatic Thoughts Questionnaire, and the Beck Depression Inventory. Family cohesion was found to be related to the degree of negative thoughts and depressive mood of the adolescents, but perceived control within the family was not. PMID- 11407638 TI - The relational self-concept scale: a context-specific self-report measure for adolescents. AB - This paper describes an alternative approach to measuring the self that directly accounts for the way individuals ruminate on their external actions in order to inform and maintain their self-image. This was achieved by designing the Relational Self-Concept Scale (RSCS), a measure that explores the role and impact that different social encounters within and around the school context have upon self-concept formation. Analysis of responses to this context-specific self report measure, obtained from a large sample of adolescents (N = 978), confirmed that the scale is multidimensional, possesses appropriate psychometric properties, and contains a high degree of ecological validity. PMID- 11407639 TI - On the excited-state energy transfer between tryptophan residues in proteins: the case of penicillin acylase. AB - The problem, whether excited-state energy transfer occurs between Trp residues in a multi-tryptophan proteins and if it does, what kind of changes it induces in different parameters of protein fluorescence, is currently under active investigation. In our previous paper [Biophys. Chem. 72 (1998) 265], the energy transfer was found and studied in detail for Na,K-ATPase. It was shown that this transfer influences all parameters of fluorescence emission, which is detected at site-selective conditions (red-edge of excitation, blue and red edges of emission). Present experiments were performed on unusually tryptophan-rich protein, bacterial penicillin acylase (28 Trp per dimer of 82 kDa) and were aimed to extend these observations. They demonstrate substantial heterogeneity in the environments of tryptophan residues within the protein structure. This suggests, that in the present case, if the energy transfer exists, it should be directed from short-wavelength-emitting to long-wavelength-emitting tryptophan residues and thus could be easily observed by a number of time-resolved and steady-state fluorescence techniques. Unexpectedly, no signature of inter-tryptophan energy transfer was found. PMID- 11407640 TI - Changes in 13C NMR chemical shifts of DNA as a tool for monitoring drug interactions. AB - The antibiotic drug, netropsin, was complexed with the DNA oligonucleotide duplex [d(GGTATACC)]2 to explore the effects of ligand binding on the 13C NMR chemical shifts of the DNA base and sugar carbons. The binding mode of netrospin to TA rich tracts of DNA has been well documented and served as an attractive model system. For the base carbons, four large changes in resonance chemical shifts were observed upon complex formation: -0.64 ppm for carbon 4 of either Ado4 or Ado6, 1.36 ppm for carbon 2 of Thd5, 1.33 ppm for carbon 5 of Thd5 and 0.94 for carbon 6 of Thd5. AdoC4 is covalently bonded to a heteroatom that is hydrogen bonded to netropsin; this relatively large deshielding is consistent with the known hydrogen bond formed at AdoN3. The three large shielding increases are consistent with hydrogen bonds to water in the minor groove being disrupted upon netropsin binding. For the DNA sugar resonances, large changes in chemical shifts were observed upon netropsin complexation. The 2', 3' and 5' 13C resonances of Thd3 and Thd5 were shielded whereas those of Ado4 and Ado6 were deshielded; the 13C resonances of 1' and 4' could not be assigned. These changes are consistent with alteration of the dynamic pseudorotational states occupied by the DNA sugars. A significant alteration in the pseudorotational states of Ado4 or Ado6 must occur as suggested by the large change in chemical shift of -1.65 ppm of the C3' carbon. In conclusion, 13C NMR may serve as a practical tool for analyzing structural changes in DNA-ligand complexes. PMID- 11407641 TI - Osmotic perturbations induce differential movements in the core and periphery of proteins, membranes and micelles. AB - Polymeric structures, namely, micelles, membranes and globular proteins share the property of two distinct regions: a hydrophobic core and a hydrophilic exterior. The dynamics of these regions of the polymeric structures were probed using selective fluorophores 1,6-diphenyl-1,3,5-hexatriene (DPH) and 1 anilinonaphthalene-8-sulfonate (ANS), respectively. Perturbation of the polymers by external osmotic pressure, ionic strength and temperature was monitored in the two regions using steady state measurements of fluorescence intensity and anisotropy. While the fluorescence lifetime of DPH and ANS did not change significantly, parallel change in steady state anisotropy values and the rotational correlation time indicated mobility in the probe/probe-domain. Osmotic perturbation of the polymers in electrolyte media led to decreased DPH mobility. Enhanced ellipticity at 222 nm in bovine serum albumin was observed in 1.5 M NaCl and sucrose media. ANS exhibited a decreased anisotropy with progressive dehydration in proteins in NaCl media, in dimyristoylphosphatidylcholine (DMPC) vesicles in sucrose media, and in neutral laurylmaltoside micelles in both NaCl and sucrose media. Thus, ANS showed responses opposite to that of DPH in these systems. A comparison with several domain selective probes indicated that DPH reported findings common to depth probes while ANS reported data common to interfacial probes used for voltage monitoring. PMID- 11407642 TI - Minimum entropy production in photosynthesis. AB - In this paper a flux-coupling model of photosynthesis is presented. By requiring minimum entropy production, it is found that the photosynthetic efficiency is essentially given by the square root of D/lambda. D and lambda are the diffusion coefficient and thermal conductivity of the rate-limiting processes in the chloroplast, respectively. For experimental values of D and lambda, the efficiency is found to be 2.4-7.5%, with a likely value of 6.1%, whereas C4 plants are known to have an efficiency of 6.2%. We conclude that the process of photosynthesis is in quantitative agreement with the principle of minimum entropy production. PMID- 11407643 TI - Counterion condensation as saturation effect under the influence of ion hydration. AB - Polyelectrolyte solutions are often described by structural theories. These theories in some cases yield values for the counterion concentration at the charged monomer surface that exceed the saturation concentration. This means a change of the ion properties due to ion immobilization or ion condensation in close vicinity to the polymer chain. The extent of this counterion condensation (CIC) and the respective surface potential are calculated from the saturation concentrations of the electrolyte involved including the influence of ion hydration on the effective dielectric number. In this paper, we shall consider all these influences by a fundamental differential equation and a set of explicit formulae yielding quantitative expressions without linearization. All calculations are based on the abstraction of an idealized elementary cell. PMID- 11407644 TI - Negative cooperativity and aggregation in biphasic binding of mastoparan X peptide to membranes with acidic lipids. AB - The change of Trp fluorescence intensity when large vesicles with 10% acidic lipid are added to mastoparan X solutions reflects a fast and a slow binding process. By means of a novel procedure of data analysis that takes advantage of so-called mass conservation plots we have separated association isotherms related to: (i) the apparent fast pre-equilibrium; and (ii) the final equilibrium, respectively. This approach also reveals that the intrinsic fluorescence signal of the slow binding is considerably raised against that of the fast binding, presumably indicating a penetration of bound peptide from the lipid/water interface into the apolar lipid core. The shape of either binding curve discloses a pronounced tendency of aggregation. Furthermore, it turns out that in the slow process the final binding ratio decreases markedly compared with the initial fast binding ratio. Accordingly the occupation of final binding sites must exert a substantial effect of negative cooperativity on the affinity of the interfacial binding states. PMID- 11407645 TI - Two types of urate binding sites on hemocyanin from the crayfish Astacus leptodactylus: an ITC study. AB - The oxygen binding behaviour of hemocyanins from Crustacea is regulated by small organic compounds such as urate and L-lactate. We investigated the binding characteristics of urate and the related compound caffeine to the 2 x 6-meric hemocyanin of A. leptodactylus under fully oxygenated conditions employing isothermal titration calorimetry (ITC). An analysis of urate and caffeine binding based on a model of n identical binding sites resulted in approximately four binding sites for caffeine and eight for urate. This result suggests that the binding process for these effectors is more complex than this most simple model. Therefore, we introduced a number of alternative models. Displacement experiments helped to select the appropriate model. Based on these experiments, at least two different types of binding sites for urate and caffeine exist on the 2 x 6-meric hemocyanin of A. leptodactylus. The two binding sites differ strongly in their specificity towards the two analogues. It can be hypothesized that two different subunit types (beta and gamma) are responsible for the two types of binding sites. PMID- 11407646 TI - The catalytic efficiency of soybean lipoxygenase-1 is enhanced at low gravity. PMID- 11407647 TI - Prevalence of silent celiac disease in patients with autoimmune thyroiditis from Northern Sardinia. AB - Celiac disease (CD) is frequently associated with other autoimmune diseases such as Type 1 diabetes mellitus, autoimmune thyroiditis (AT), and Addison's disease. The frequency of these associations varies with the populations studied. We conducted this study to ascertain the prevalence of CD in patients with AT from Sardinia, an area with a very high prevalence of CD. To this aim, 297 consecutive patients with AT (as defined by elevated antithyroid antibody levels and a positive ultrasound scan) were studied. Immunoglobulin A and G-class antigliadin antibodies were assayed in serum; if either or both were positive, antiendomysium antibodies were determined. If two markers were positive, serum ferritin, folate, and vitamin B12 levels were measured and jejunal biopsy was suggested. Thirteen out of the 14 patients who showed at least two positive markers consented to jejunal biopsy and all of them showed histological features of CD. The prevalence of CD in AT patients was 4-fold greater than that observed in the general population (4.37 vs 1.06%, p<0.0001). Ferritin was low in 6 and vitamin B12 in 2 out of 13 patients; serum folates were normal in all patients. Molecular typing of HLA class II alleles showed an increased frequency of the extended haplotype DRB1*0301/DQA1*0501/DQB1*0201. None of our patients had a history of gastrointestinal symptoms. We confirm the increased prevalence of silent CD in patients with AT. Patients with AT ought to be regarded as a high-risk group for CD and should be screened routinely for it; if negative, screening tests should be repeated at regular intervals. PMID- 11407648 TI - Growth hormone responses to oral glucose and intravenous thyrotropin-releasing hormone in acromegalic patients treated by slow-release lanreotide. AB - The aim of this study was to assess GH response to oral glucose tolerance test (OGTT) and TRH stimulation test in a group of 10 patients with active post operative acromegaly before and after long-term slow-release (SR) lanreotide therapy (30 mg im every 10-14 days). Seven patients (2 males, 5 females, 29-71 yr), who during therapy maintained plasma GH and IGF-I concentrations under 5 microg/l and 450 microg/l, respectively, were considered as responders and studied for 24 (1 patient) to 36 months (6 patients). Three patients (1 male, 2 females, 46-61 yr) with levels of GH and IGF-I above those values were studied for 12 months. The OGTT (75 g po) and TRH test (400 microg iv) were repeated before and after 6, 12, 24 and 36 months. The GH response to OGTT was abnormal (nadir: >2 microg/l) at 6 and 12 months in poorly responsive patients. This response was normalized in all responsive patients. Nonetheless, 2 responsive patients showed abnormal GH values after OGTT once each throughout the 36-month study period. The GH response to TRH was characterized by great variability and exhibited unpredictable behavior throughout the study period both in responsive and in poorly responsive patients. Only 2 patients in the responsive group showed persistent normal GH levels (peak: < or =5 microg/l) after TRH for 3 yr. In conclusion, SR lanreotide treatment gave rise to a correct control of GH hypersecretion and to a normalization of GH response to oral glucose in 7 out of 10 patients, although it did not abolish the paradoxical reaction of GH to TRH in all responders. The effect of SR lanreotide on GH response to glucose tolerance test was not paralleled by GH response to TRH. PMID- 11407649 TI - Neuroendocrine effects of subcutaneous sumatriptan in patients with migraine. AB - We evaluated the sensitivity of 5-HT1D receptors in patients with migraine using sumatriptan as a pharmacological probe. The drug inhibits the release of ACTH, cortisol and prolactin and this effect may be used to explore the function of serotoninergic systems in vivo. We administered sumatriptan (6 mg sc) and placebo to 15 migraineurs, during the headache-free period, and to 10 healthy controls. Blood samples were collected -15, 0, 15, 30, 45, 60 and 90 min after injections. Sumatriptan induced a significant (p<0.01) decrease of ACTH, cortisol and prolactin concentrations both in patients with migraine and in controls. The neuroendocrine response was not significantly different in the two groups. Our results suggest that 5-HT1D receptor sensitivity is not altered in migraine. PMID- 11407650 TI - Increased prevalence of primary hyperparathyroidism in treated breast cancer. AB - Hypercalcemia occurring in patients with advanced breast cancer (BC) is generally due to osteolytic metastases or to the activity of circulating tumor-derived products. In these conditions, the production of endogenous PTH is reduced. The frequency of hypercalcemia due to primary hyperparathyroidism in breast cancer is unknown. We examined the occurrence of primary hyperparathyroidism in a large group of women with treated BC. A total of 100 consecutive women aged 28-80 years with treated breast cancer were enrolled. One hundred and two healthy age-matched women and 60 age-matched female patients with differentiated thyroid carcinoma examined before thyroidectomy were used as controls. Intact serum PTH and serum calcium were measured in all patients and controls. Hypercalcemia associated with elevated serum PTH concentration indicating primary hyperparathyroidism was found in 7 BC patients (7%) and in none of healthy women or patients with thyroid cancer. The pre-operative staging of BC patients with primary hyperparathyroidism was I in six and II in one of them, and no patient had evidence of distant metastases. A parathyroid adenoma was found in all 6 BC patients submitted to neck exploration, one patient refused surgery. Serum calcium and PTH concentrations returned to normal levels after surgery. Two BC patients had increased serum PTH and normal calcium concentrations. One of them had low serum 25-hydroxyvitamin D [25(OH)D]. One patient with spread bone metastases had neoplastic hypercalcemia with undetectable serum PTH concentration. All remaining 90 BC patients had serum calcium and PTH concentrations within normal limits, but their mean (+/-SD) values (9.6+/-0.5 mg/dl for serum calcium, 38.0+/-16.4 mg/dl for serum PTH ) were slightly but significantly greater than in normal controls (9.3+/-0.5 mg/dl, p=0.003 and 27.9+/-10.6 pg/ml, p=0.0001, respectively) and in patients with thyroid cancer (9.2+/-0.6 mg/dl, p=0.001 and 26.2+/-11.0 pg/ml, p=0.001), with no relationship with clinical staging or anti-tumor therapy. IN CONCLUSION: 1) an increased frequency of parathyroid adenoma was found in BC patients with mildly aggressive neoplastic disease; 2) in BC patients with no evidence of primary hyperparathyroidism mean serum PTH and calcium concentrations were significantly greater than in healthy controls and in patients with thyroid carcinoma; and 3) this finding was unrelated to clinical staging or anti-tumor therapy. Thus, primary hyperparathyroidism should be considered as a possible cause of hypercalcemia in patients with non-aggressive breast cancer. We suggest that serum PTH should be determined in all BC patients with increased serum calcium concentration, especially in those with no evidence of metastatic disease. PMID- 11407651 TI - Non-hyperfunctioning nodules from multinodular goiters: a minor role in pathogenesis for somatic activating mutations in the TSH-receptor and Gsalpha subunit genes. AB - Constitutive activation of the cAMP pathway stimulates thyrocyte proliferation. Gain-of-function mutations in Gsalpha protein have already been identified in thyroid nodules which have lost the ability to trap iodine. In contrast, most of the studies failed to detect somatic activating mutations in the thyrotropin receptor (TSH-R) in non-hyperfunctioning thyroid tumors. The aim of this study was to screen for mutations TSH-R exon 10, encoding the whole intracytoplasmic area involved in signal transduction, and Gsalpha exons 8 and 9, containing the two hot-spot codons 201 and 227, in a subset of non-hyperfunctioning nodules from multinodular goiter. Identified by matching ultrasonography and scintiscan, 22 eufunctioning (normal 99Tc uptake) and 15 nonfunctioning (decreased 99Tc uptake) nodules from 27 non-toxic multinodular goiters were isolated. After DNA extraction, TSH-R exon 10 was analyzed by direct sequencing of the PCR products and Gsalpha exons 8 and 9 by Denaturing Gradient Gel Electrophoresis. No mutation of TSH-R or Gsalpha was detected in the 37 nodules analyzed. This absence of mutation, despite the use of two sensitive screening methods associated with the analysis of the TSH-R whole intracytoplasmic area and Gsalpha two hot-spot codons, suggests that TSH-R and Gsalpha play a minor role in the pathogenesis of non-toxic nodules from multinodular goiters. PMID- 11407652 TI - Age-related changes in bone density among healthy Greek males. AB - Osteoporosis in men is increasingly recognized as a problem in clinical medicine, but it has received much less attention than its counterpart in women. It is termed idiopathic if no known cause of bone disease can be identified clinically or in the laboratory. The true incidence of idiopathic osteoporosis (IO) in males is difficult to estimate because population characteristics and referral patterns differ so widely. The aim of this study was to investigate the incidence of IO in healthy Greek male volunteers by measuring bone mineral density (BMD) at four skeletal sites and examining the relations among age, BMI, and bone status. This type of information has not yet been published. We considered osteoporosis to be present when the BMD was less than or equal to -2.5 SD from the average value for healthy young men. Three hundred and sixty-three normal male volunteers were investigated. The mean age was 51.3+/-8.7 yr, and BMI was 27.5+/-3.7 kg/m2. In all subjects BMD at four skeletal sites - lumbar spine (LS), femoral neck (FN), Ward's triangle (WT), and finally trochanter (T) - was measured using dual-energy X-ray absorptiometry (DEXA). T-score, Z-score and g/cm2 values were estimated. Forty-four subjects (11%) had BMD< or =-2.5 SD (T-score). The mean age and BMI for the men with decreased BMD was 54.8+/-6.4 yr and 26.3+/-3.3 kg/m2, whereas mean age and BMI for those with normal BMD was 51.0+/-8.9 yr and 27.6+/-3.6 kg/m2, respectively. These differences were statistically significant (p<0.001 and p<0.05, respectively). A positive correlation was found between BMI and bone density (g/cm2) at three skeletal sites: LS (r=0.235, p<0.001), WT (r=0.126, p<0.001) and FN (r=0.260, p<0.001). A positive correlation was also found between BMI and T-score at all skeletal sites studied: LS (r=0.276, p<0.001), WT (r=0.133, p<0.05), FN (r=0.233, p<0.001), and T (r=0.305, p<0.001). Finally, a positive correlation was also found between BMI and Z-score: LS (r=0.256, p<0.001), WT (r=0.117, p<0.005), FN (r=0.240, p<0.001), and T (r=0.187, p<0.001). A negative correlation was found between age and bone density (g/cm2) at FN (r= 0.157, p<0.01) and WT (r=-0.183, p<0.001). The same was true between age and T score at FN only (r=0.137, p<0.05). Furthermore, a similar correlation was found between age and Z-score at LS (r=0.174, p<0.001). When ANOVA one-way analysis was used, a significant difference was found between the different age groups and BMD (g/cm2) at FN, T, and WT (p<0.001 for all sites). For T-score, a significant difference between age groups was found only at FN (p<0.005). Finally, a significant difference in Z-score was found at FN (p<0.001) and LS (p<0.005). When multiple regression analysis was applied, it was found that BMD (g/cm2) at two sites, FN and WT, independently correlated with age and BMI (FN: p<0.001 for both, WT: p<0.01 and p<0.05, respectively). Finally, we found an accelerated trend toward decreased BMD (g/cm2), when the odds ratio was applied. In conclusion, this study demonstrated that 11% of otherwise healthy Greek men had BMD less than or equal to -2.5 SD. A strong association was found between BMD (g/cm2) and age at three skeletal sites when ANOVA one-way analysis was applied. Moreover, BMD was positively correlated with BMI and negatively correlated with age. Currently available data are sparse and much more research is needed to increase our understanding concerning the etiology of this condition as well as illuminating the relationship between bone density and fracture. PMID- 11407653 TI - Urinary excretion pattern of catecholestrogens in preovulatory LH surge during the 4-day estrous cycle of rats. AB - The formation of catecholestrogens represents a major pathway of estrogen metabolism and catecholestrogens are regarded as the main estrogen metabolite in non-pregnant state of various mammalian systems. In the present investigation, level of 2-hydroxyestrone, the major catecholestrogen excreted in rat urine, was measured by radioimmunoassay following acid hydrolysis and column chromatography of the 24-h urine samples of female Sprague Dawley non-pregnant rats during their 4-day estrous cycle. Urinary levels of estrone, estradiol and estriol were measured. Unlike the plasma level, urinary 2-hydroxyestrone showed a marked increase during the pre-ovulatory LH surge suggesting a plausible role of catecholestrogen in the mid-cycle elevation of the gonadotropin level in normal cycling rats. PMID- 11407654 TI - Impairment of GH responsiveness to GH-releasing hexapeptide (GHRP-6) in Prader Willi syndrome. AB - The aim of this study was to evaluate the GH-releasing activity of a synthetic hexapeptide, GHRP-6, in the Prader-Willi syndrome (PWS). Sixteen PWS patients (7 males and 9 females, aged 12.7-38.3 yr), 15 with essential obesity (OB) (7 males and 8 females, aged 12.9-42.9 yr), and 8 short normal children (SN; 3 males and 5 females, aged 10.2-14.3 yr) underwent 2 tests on separate occasions, being challenged with GHRP-6 (1 microg/kg, iv) or GHRH (1 microg/kg, iv)+PD (60 or 120 mg for children or adults, po). Moreover, in 11 patients with PWS and in the group of SN, the GH response to at least 2 stimulation tests had been previously determined. GH was analyzed either as mean peak values (GHp, mcg/l), or as the area under the curve (AUC, mcg/l/h) and the net incremental area under the curve (nAUC, mcg/l/h). In the group of PWS subjects, GH responses to both GHRP-6 (GHp: 11.4+/-2.0; AUC: 588+/-113; nAUC: 483+/-108) and GHRH+PD (GHp: 7.3+/-1.8; AUC: 486+/-122; nAUC: 371+/-250) were significantly lower than those observed either in OB (GHRP-6: GHp: 25.7+/-3.2, p<0.003; AUC: 1833+/-305, p<0.005; nAUC: 1640+/ 263, p<0.0001. GHRH+PD: GHp: 15.1+/-2.4, p<0.009; AUC: 1249+/-248, p<0.003; nAUC: 918+/-230, p<0.006) or in SN patients (GHRP-6: GHp: 39.1+/-3.1, p<0.0001; AUC: 2792+/-158, p<0.0001; nAUC: 2705+/-165, p<0.00005. GHRH+PD: GHp: 27.5+/-3.7, p<0.0001; AUC: 1873+/-251, p<0.0001; nAUC: 1692+/-219, p<0.0005). Unlike control groups, in PWS patients GH levels after GHRP-6 did not differ from those obtained after GHRH+PD. Interestingly, low IGF-I values were present in all PWS subjects. Furthermore, no patient with PWS showed normal GH response to the previously performed GH stimulation tests. As already reported, GH release after GHRP-6 or GHRH+PD was significantly lower in OB than in SN subjects. In conclusion, our data indicate that: 1) GH response to GHRP-6 is clearly impaired in PWS; 2) the blunted GH responses to the provocative stimuli in PWS are not an artifact of obesity; 3) short stature in PWS is caused by a complex dysfunction of the hypothalamo-pituitary structures. PMID- 11407655 TI - Familial isolated parathyroid adenoma in a consanguineous family. AB - The 23-year-old Caucasian male propositus presented with symptomatic hypercalcemia, hypophosphatemia and normocalciuria for 2 months. His 29-year-old brother had undergone an operation for recurrent parathyroid adenoma at age 26 and 28. No other member of the family was affected. His father and mother were second-degree relatives. Laboratory studies showed primary hyperparathyroidism (pHPT), while the remaining endocrine studies and genetic testing for multiple endocrine neoplasia 1 and 2A were normal. Technetium-cardiolite scintigraphy and ultrasound scans revealed a parathyroid mass at the left lower neck. Apart from bilateral hearing loss due to gentamicin treatment as a pre-term child, the patient was in of good health. Signs or symptoms of other endocrinopathies were absent. The patient was referred for parathyroidectomy with subsequent autotransplantation of the remaining glands into his sternocleidomastoid muscle. Histological examination revealed an adenoma with oncocytic differentiation, similar to that seen in his brother. The disease may follow a recessive mode of inheritance or may be due to a dominant germ-cell mutation in one of the parents. The presented case may ultimately help in elucidating the molecular genetic basis of this rare form of pHPT. PMID- 11407656 TI - False serum calcitonin high levels using a non-competitive two-site IRMA. AB - Dual site antibody-base immunoassays are commonly used in clinical laboratories to quantify the CT serum concentrations as a specific and sensitive marker of medullary thyroid carcinoma (MTC). Heterophile antibodies can interfere with these assays, however, and cause erroneous results. In order to avoid this interference, immobilized and conjugated antibodies from two different animal species or immunoreactive antibody fragments, as well as the addition of non immune globulins, are generally included among the assay reagents. We describe the case of a 73-year-old man affected by a multinodular goiter, who showed high basal CT plasma levels as measured by a monoclonal antibody based IRMA. The finding of negative results for the presence of MTC at fine needle aspiration (FNA) and the mild increase observed in plasma CT during a pentagastrin (Pg) stimulation test, suggested that the high CT levels might depend on a cross reaction with heterophilic antibodies. In fact, after the addition of the heterophilic blocking tube (HBT) to each specimen, the CT levels markedly decreased by more than 80% (average decrease+/-SE= 87.6+/-2.668%). Such a decrease strongly suggests that in our case the routinely used F(ab')2 fragments were unable to eliminate all of the interference and that the elevated serum CT levels might have been caused by human heterophilic antibodies. In conclusion, these results indicate a novel cause of CT false positivity, suggesting that high serum CT levels, when combined with a slight increase during Pg stimulation, should be critically interpreted in view of the possible presence of heterophilic antibodies in the specimens. PMID- 11407657 TI - A case of spurious hypercalcitoninemia: a cautionary tale on the use of plasma calcitonin assays in the screening of patients with thyroid nodules for neoplasia. AB - The measurement of plasma CT has an important role as a screening test for medullary thyroid carcinoma (MTC) in patients with thyroid nodules. However, elevated plasma CT levels should be interpreted within the context of the overall clinical picture in each individual case and carefully validated before therapeutic decisions are made. We present the case of a 17-yr-old girl who was referred to us with a thyroid nodule and elevated plasma CT levels, as measured by a one-site RIA not involving prior plasma extraction. Plasma CT was re measured using two different methods, a RIA with prior plasma extraction and a two-site immunochemiluminometric assay (ICMA), and was either very low or undetectable. Subsequently, samples were re-assayed using the initially applied CT RIA; plasma CT levels were again found to be elevated. These elevations were of a spurious nature, probably caused by the presence of an unidentified substance in the patient's plasma interfering with the measurement of CT in the initially used RIA. Our patient was eventually diagnosed with Hashimoto's thyroiditis, and had no evidence of MTC. As several conditions can cause either true or spurious hypercalcitoninemia, we suggest that elevated plasma CT levels should be confirmed at least once before other extensive diagnostic investigations are initiated or thyroidectomy is recommended. Finally, the assay selected should detect only the mature CT molecule. PMID- 11407659 TI - Magnetic resonance imaging: a complementary tool in the evaluation of thyroid nodules? PMID- 11407658 TI - Clinical genetics of multiple endocrine neoplasias, Carney complex and related syndromes. AB - The list of multiple endocrine neoplasias (MENs) that have been molecularly elucidated is growing with the most recent addition of Carney complex. MEN type 1 (MEN 1), which affects primarily the pituitary, pancreas, and parathyroid glands, is caused by mutations in the menin gene. MEN type 2 (MEN 2) syndromes, MEN 2A and MEN 2B that affect mainly the thyroid and parathyroid glands and the adrenal medulla, and familial medullary thyroid carcinoma (FMTC), are caused by mutations in the REToncogene. Finally, Carney complex, which affects the adrenal cortex, the pituitary and thyroid glands, and the gonads, is caused by mutations in the gene that codes for regulatory subunit type 1A of protein kinase A (PKA) (PRKAR1A) in at least half of the known patients. Molecular defects have also been identified in syndromes related to the MENs, like Peutz-Jeghers syndrome (PJS) (the STK11/LKB1 gene), and Cowden (CD; the PTEN gene) and von Hippel-Lindau disease (VHLD; the VHL gene). Although recognition of these syndromes at a young age generally improves prognosis, the need for molecular testing in the diagnostic evaluation of the MENs is less clear. This review presents the newest information on the clinical and molecular genetics of the MENs (MEN 1, MEN 2, and Carney complex), including recommendations for genetic screening, and discusses briefly the related syndromes PJS, CD and VHLD. PMID- 11407660 TI - The crucifixion of Saint Andrew. PMID- 11407661 TI - Re-evaluation of the thyroxine binding to human plasma lipoproteins using three techniques. AB - Using gel filtration chromatography (GFC), we found that human VLDL, LDL and HDL transport approximately 0.03, 0.2 and 3% of plasma T4. As T4 could dissociate from carriers during GFC, we evaluated [125I]T4 transport to lipoproteins (Lp) in 50 normolipidemic and euthyroid subjects by GFC and 2 independent methods: zone electrophoresis (ZE) and radioimmunoprecipitation (RIP). At GFC, VLDL+LDL transported less T4 (approximately 0.3%) than either ZE (approximately 1.3%) or RIP (approximately 1.5-2.0%). In contrast, GFC values for the HDL (approximately 3%) were close to the RIP values (approximately 3.5%), but lower than ZE (approximately 8.5%) because of partial co-migration with TBG. In conclusion, GFC underestimates T4 binding to VLDL and LDL, which, indeed, is of the same magnitude as binding of some steroid hormones to CBG and SHBG. Hence, the anti atherosclerotic effects of T4 resulting from binding to the LDL should be greater than anticipated based on GFC data. PMID- 11407662 TI - Long-acting nitrates in portal hypertension: to be or not to be? AB - The major complication of portal hypertension is represented by gastrointestinal haemorrhage from ruptured oesophageal varices. Gold standard prophylaxis with non selective beta-blockers is able to decrease the risk of bleeding or rebleeding only in a fraction of patients, thus additional forms of treatment are under investigation. Long-acting nitrates have been considered the best candidates to improve the pharmacological response. The rationale for the use of nitrates in portal hypertension is primarily based on the fact that they lead to a decrease in the hepatic venous pressure gradient and on the knowledge that deficient intrahepatic nitric oxide release could be one of the mechanisms involved in the development of increased portal resistance in early cirrhosis. Ten randomised controlled trials have, so far, investigated the clinical usefulness of long acting nitrates in portal hypertension. Five of them explored the field of primary prophylaxis and the others, the use of nitrates in the prevention of rebleeding. The results of these randomised controlled trials are partially contradictory as far as concerns prevention of bleeding or rebleeding, survival and treatment-related complications. A common finding emerging from most of these studies suggests that the potential for a beneficial or detrimental effect of nitrates depends on the stage of liver disease and the extension of portal collaterals. Thus, in the early stage of cirrhosis, it would be desirable to target nitrates to the liver microvasculature, while, in a later stage, nitrates could be deleterious by aggravating the hyperdynamic syndrome through the expansion of the vascular bed. Whether or not nitrates may have a role in the primary and/or secondary prophylaxis of bleeding needs to be addressed in further long-term studies. PMID- 11407663 TI - Current diagnosis and management of gastrointestinal neuroendocrine tumours. PMID- 11407664 TI - Helicobacter pylori as the cause of coronary artery restenosis following angioplasty--is the way to a man's heart disease through his stomach? PMID- 11407665 TI - Digestive neuroendocrine tumours: diagnosis and treatment in Italy. A survey by the Oncology Study Section of the Italian Society of Gastroenterology (SIGE). AB - BACKGROUND: New insights in the diagnosis and treatment of digestive neuroendocrine tumours have prompted a renewed interest in these rare and complex diseases. AIM: To establish how many new cases of digestive neuroendocrine tumours were diagnosed, and how they were treated, at gastroenterological centres across Italy during a two-year period (1997-1998). METHODS: The 12 centres taking part filled in a data collection form reporting type of tumour, methods of diagnosis and therapeutic strategies adopted in each case. Data were collected and analysed by the authors of the present report. RESULTS: Data refer to 98 patients, 22 with functioning and 76 with non-functioning digestive neuroendocrine tumours [50 carcinoids, 48 pancreatic endocrine tumour syndromes]. Primary tumours were localised in 96% (38% with metastases) of non-functioning and 81% (50% with metastases) of functioning tumours. These were surgically removed in >80% of patients in both groups. Somatostatin analogue treatment, with or without interferon, was administered in 35% of patients, while chemotherapy was used in 9% and 23% of functioning and non-functioning tumours, respectively. The imaging study always included a computed tomography scan (20% helical computed tomography). Magnetic resonance and somatostatin receptor scintigraphy were also performed, the former in 41% and 21% of the two (functioning and non functioning tumour) groups, the latter in 45% and 30%. CONCLUSIONS: The number of functioning digestive neuroendocrine tumours reported was lower than expected. Surgery plays a major role in the treatment of these tumours in all centres. Overall, in only a small number of patients was a multidisciplinary approach applied. PMID- 11407666 TI - Prevalence of Helicobacter pylori infection in coronary artery disease and effect of its eradication on coronary lumen reduction after percutaneous coronary angioplasty. AB - BACKGROUND: Gastric infection caused by Helicobacter pylori has recently been associated with increased risk of coronary artery disease. AIM: To: 1) determine seroprevalence of Helicobacter pylori and its cytotoxin associated gene A in patients with/without coronary artery disease (group A), 2) assess influence of Helicobacter pylori eradication on coronary artery lumen reduction after percutaneous coronary angioplasty (group B) and 3) determine influence of Helicobacter pylori eradication on plasma cytokines, lipids and coagulation factors in patients subjected to percutaneous coronary angioplasty (group B). PATIENTS AND METHODS: Group A included 100 patients with coronary artery disease (subgroup 1) and 100 patients without (subgroup II). For Helicobacter pylori seroprevalence, plasma anti-Helicobacter pylori and anti-cytotoxin associated gene A IgG were examined. Group B included 40 patients with significant single vessel coronary arterial disease and Helicobacter pylori infection confirmed by 13C-urea breath test and serologically using anti-Helicobacter pylori and anti cytotoxin associated gene A IgG. Six months after percutaneous coronary angioplasty and triple anti-Helicobacter pylori therapy, the Helicobacter pylori status reassessed by urea breath test was negative in all but two patients of subgroup I subjected to Helicobacter pylori therapy. Coronary angiography and laboratory tests were repeated in both subgroups of group B included in the trial and reduction in coronary artery lumen in these subgroups was compared to baseline after percutaneous coronary angioplasty considered as 100%. RESULTS: Helicobacter pylori seropositivity reached 81.5% of coronary artery disease (subgroup I) and was significantly higher than that in controls without coronary artery disease (subgroup II) (51%), the odds ratio being 4.3 for Helicobacter pylori in coronary artery disease. Cytotoxin associated gene A IgG detection was also significantly higher (47.3%) in coronary artery disease than in controls (28%) giving the odds ratio about 2.3. Mean coronary artery lumen reduction in patients undergoing percutaneous coronary angioplasty + Helicobacter pylori eradication therapy (subgroup I) was significantly (p<0.05) smaller compared to percutaneous coronary angioplasty + placebo-treated subgroup II (22% vs 41%). CONCLUSIONS: 1) There is a significant link between coronary artery disease and infection with Helicobacter pylori, especially expressing CagA proteins, 2) Helicobacter pylori eradication significantly attenuates reduction in coronary artery lumen in coronary artery disease patients after percutaneous coronary angioplasty possibly by elimination of chronic inflammation and decline in proinflammatory cytokine release, and 3) Infection of Chlamydia pneumoniae in these percutaneous coronary angioplasty patients is not affected by eradication therapy. PMID- 11407667 TI - Efficacy and safety of Nd:YAG laser for the treatment of bleeding from radiation proctocolitis. AB - BACKGROUND: Bleeding due to radiation proctocolitis is a frequent and severe complication of radiotherapy in cancers of the pelvis. AIM: The aim of this study was to evaluate the efficacy and safety of endoscopic treatment with Nd:YAG laser in this condition. PATIENTS AND METHODS: A series of 9 patients with radiation induced damage in the rectum and sigma were treated with endoscopic Nd:YAG laser until significant bleeding stopped and endoscopic features of proctocolitis improved. They received a median of 3 laser treatments (range 1-10) over a maximum time period of 11 months. RESULTS: In 4 cases, bleeding ceased and, in 4, it was reduced to occasional spotting. In the remaining patient, laser therapy led to only a transient improvement, but did not modify the requirement of blood transfusion. In the 5 patients also suffering from urgency, incontinence and/or rectal mucoid discharge, the laser therapy course also relieved these symptoms. No significant treatment-related complications were observed. CONCLUSIONS: Endoscopic Nd: YAG laser is a useful and safe treatment for patients with bleeding due to radiation proctocolitis. PMID- 11407668 TI - Lactose malabsorption, irritable bowel syndrome and self-reported milk intolerance. AB - BACKGROUND: The relationship between lactose malabsorption, irritable bowel syndrome and development of intestinal symptoms is unclear, especially when the ingested dose of milk is small. Thus, the role of hydrogen breath testing in the diagnostic work-up of patients with nonspecific intestinal symptoms is still debated. AIMS: To establish the relationship between lactose malabsorption, severe self-reported milk intolerance, irritable bowel syndrome and related symptoms. METHODS: The prevalence of lactose malabsorption was prospectively assessed by means of a hydrogen breath test in 839 patients (503 with irritable bowel syndrome, based on the Rome criteria, regularly consuming milk, and 336 subjects who identified themself as milk intolerant, after an oral load of 25 g lactose). The test was considered "positive" when a hydrogen peak exceeding 20 ppm over baseline values was observed in two or more samples. Attempts were also made to establish whether the predominant presenting symptom (diarrhoea, constipation, alternating diarrhoea and constipation, pain and gaseousness) might be helpful in predicting the outcome of the breath test. RESULTS: The prevalence of a positive breath test was comparable in the two groups (337 patients with irritable bowel syndrome (66.9%) vs 240 patients with milk intolerance (71.4%)). The same holds true for the first peak of hydrogen excretion, total hydrogen output and prevalence of symptoms during, and in the four hours after, the test. The predominant presenting symptom was not useful for predicting outcome of the test either in regular milk users or in milk intolerant subjects. CONCLUSIONS: The almost identical results of the lactose breath test of patients with irritable bowel syndrome and subjects with self-reported milk intolerance suggests that the two conditions overlap to such an extent that the clinical approach should be the same. A lactose breath test should always be included in the diagnostic work-up for irritable bowel syndrome, as fermentation of malabsorbed lactose is likely responsible for triggering symptoms. Conversely, lactase deficiency is probably irrelevant in most subjects not affected by irritable bowel syndrome, within a moderate milk consumption. PMID- 11407669 TI - Interactions between metabolic disorders (diabetes, gallstones, and dyslipidaemia) and the progression of chronic hepatitis C virus infection to cirrhosis and hepatocellular carcinoma. A cross-sectional multicentre survey. AB - BACKGROUND: Diabetes, gallstones and dyslipidaemia are widespread, metabolically related, disorders that can affect the liver, often in a clinically silent fashion. AIM: To investigate whether the presence of these disorders may worsen chronic viral disease by inducing additional liver damage, revealed by variations in serum increases of aminotransferase, alkaline phosphatase and gamma-glutamyl transpeptidase activities. PATIENTS AND METHODS: This retrospective, cross sectional study involved 1,195 patients with chronic hepatitis C virus infection: 47.2% chronic hepatitis, 45.2% cirrhosis, and 7.6% hepatocellular carcinoma. 14.9% of patients had enzymatic cholestasis, defined as combined increase of alkaline phosphatase and gamma-glutamyl-transpeptidase. A Log-linear statistical model was applied to the following variables: stages of liver disease, diabetes, cholelithiasis, hypertriglyceridaemia, hypercholesterolaemia, and enzymatic cholestasis. RESULTS: Log-linear analysis, applied to categorical variables, revealed, for the first time, a three-way interaction between the stages of chronic liver disease, diabetes, and enzymatic cholestasis. Two-way interactions demonstrated that liver disease stages correlated directly to the prevalence of cholelithiasis and inversely to hypercholesterolaemia. Irrespective of the liver disease stage, hypertriglyceridaemia correlated to hypercholesterolaemia. CONCLUSIONS: This study discloses a synergistic liver damaging effect of diabetes and hepatitis C virus. The three-way interaction obtained by our analysis suggests that diabetes is a risk factor for the progression of viral liver disease and that it contributes to disease evolution, at least in part, by induction of cholestasis. PMID- 11407670 TI - Interferon-related thyroid autoimmunity and long-term clinical outcome of chronic hepatitis C. AB - BACKGROUND: A high incidence of thyroid autoantibodies and/or disorders was observed in subjects with hepatitis C virus-related chronic hepatitis during interferon-alpha therapy. AIM: To evaluate whether thyroid autoimmunity and dysfunction, induced by interferon-alpha therapy, could be viewed as predictors for treatment response and as valid prognostic markers of liver disease progression. PATIENTS: A total of 136 subjects (96 males/40 females; median age 48 years; range 23-64) affected by biopsy-proven chronic hepatitis C (33.1% with compensated liver cirrhosis). METHODS: All subjects were treated with interferon alpha therapy at 6 MU 3 times weekly for 12 months and then followed up for an average period of 60 months (range 12-108). Routine laboratory tests, virological assessment, liver ultrasound, thyroid function tests (serum free triiodothyronine, free-thyroxine, serum thyrotropin), and autoimmunity were performed for all subjects. RESULTS: Percentage of thyroid autoimmunity and thyroid dysfunction in long-term responders was not significantly different compared to that in non-responders (47.0% and 11.8% vs 35.3% and 5.9%, respectively; non significant). The multivariate model demonstrated that the absence of cirrhosis was the only factor significantly related to successful response to therapy (odds ratio: 14.9; 95% confidence interval: 1.9-115.0 for chronic hepatitis C vs presence of cirrhosis). Moreover, the occurrence of thyroid autoimmunity during interferon therapy was similar both in patients with or without worsening of liver disease (33.3% and 39.8%, respectively; p = not significant). No subject with on-going liver disease developed thyroid dysfunction during treatment, as opposed to the 10/118 (8.4%) with a better course of liver disease; however, this difference was not statistically significant. The multivariate model showed that age was the only covariate significantly associated with unfavourable outcome of liver disease (odds ratio: 18.6; 95% confidence interval: 2.3-151.9, for those over 48 years vs younger patients). CONCLUSIONS: There is no evidence that the immune mechanism involved in the pathogenesis of thyroid autoimmune phenomena is the same as that regulating the therapeutic clearance of HCV or modulating the unfavourable course of HCV-related chronic hepatitis. However, our study confirmed that liver disease seems to progress more slowly in younger subjects. PMID- 11407671 TI - Primary melanoma of the oesophagus treated endoscopically. A case report. AB - Primary melanoma is a rare neoplasm of the oesophagus, with dismal outcome in most cases. We report a case of primary melanoma of the oesophagus treated endoscopically, as coexisting illness prohibited surgical resection of the tumour. A review of the literature is made. PMID- 11407672 TI - New therapeutic modalities for benign oesophageal disease: an overview. PMID- 11407673 TI - Current trends in the management of achalasia. AB - Despite the recent advances in the understanding of the pathophysiology of achalasia, aetiology remains obscure and this primary oesophageal motor disorder is still considered "idiopathic" in nature. As a consequence, the therapeutic approach remains palliative. Since there is little or no chance of improving the motor abnormalities of the oesophageal body, treatment of achalasia is aimed at symptomatic relief of functional lower oesophageal sphincter obstruction. Pharmacologic treatment induces only a limited and brief improvement. It may be used to treat early cases of achalasia without significant oesophageal dilatation and to manage patients exhibiting some but not all the characteristics of achalasia (e.g. transitional forms). In any event, drug therapy should be seen as a short-term measure and be considered as an alternative only in patients unfit to undergo pneumatic dilatation or surgery. Pneumatic dilatation and surgical myotomy (now increasingly carried out through a minimally invasive approach) remain, therefore, the two main approaches which guarantee long-lasting symptomatic relief. Unfortunately, both pneumatic dilatation and Heller cardiomyotomy are only palliative as neither reliably reverses oesophageal aperistalsis not corrects the incomplete postdeglutition relaxation of the lower oesophageal sphincter. They do, however, improve symptoms by lowering lower oesophageal sphincter pressure thus enhancing oesophageal emptying by gravity. Recently a third approach, consisting in perendoscopic injection of botulinum toxin into the lower oesophageal sphincter is gaining acceptance. Indeed, more endoscopists are finding this kind of treatment attractive because it does not carry the risk of perforation that can occur with pneumatic dilatation. However, since symptomatic improvement with botulinum toxin only lasts a few months, either repeated injections are required or the patient must be switched to other therapy. There may be, however, subsets of patients for whom BoTox injection is the preferred approach. They probably include elderly patients or patients with multiple medical problems who are poor candidates for more invasive procedures as well as those unwilling to have either surgery or pneumatic dilatation. Future approaches to achalasia may markedly change from the suggested algorithm depending on the long-term efficacy and safety as well as cost analysis of BoTox injection and of minimally invasive surgery. PMID- 11407674 TI - Present medical management of Barrett's oesophagus. AB - The management of Barrett's oesophagus should aim to treat symptoms, and prevent complications of reflux. Treatment of choice is a proton pump inhibitor, with the option of surgical treatment in younger patients. Uncertainties remain about the significance of short segment Barrett's oesophagus. Doubts also remain about the benefit and cost-effectiveness of conventional surveillance of Barrett's oesophagus; for patients with an annual cancer risk of 0.5% 5-yearly endoscopy and biopsies for patients without dysplasia represent the best strategy Novel modalities such as photodynamic therapy have shown encouraging results for high grade dysplasia or early cancer for patients unsuitable for surgery. Finally, agents such as cyclooxygenase-2 inhibitors hold promise for prevention of malignancy in Barrett's oesophagus. PMID- 11407675 TI - Variceal band ligation in the management of bleeding oesophageal varices: an overview. AB - Endotherapy is, in many centres, still the cornerstone in the first-line approach of a patient with variceal haemorrhage. Since the introduction of the multiband ligation, devices, active variceal bleeding is controlled as efficiently as with sclerotherapy, and variceal eradication can be obtained much faster with fewer treatment-related complications. This observation is now supported by several randomized trials. Combining sclerotherapy with band ligation has no additional effect and can even increase the number of side-effects. Therefore, multiband ligation can be considered as a first-step treatment for bleeding oesophageal varices. PMID- 11407676 TI - Minimally invasive surgery for benign oesophageal disease. AB - Laparoscopy and thoracoscopy in recent years, have gained increasing popularity in the treatment of many benign oesophageal diseases in referral centres. In some instances, as for example gastro-oesophageal reflux disease, minimally invasive approach has become the surgical access of first choice. In fact, in experienced hands, postoperative complications are reduced compared to open surgery, the operated patients benefit from better comfort, and the results are comparable to the traditional approach in terms of effectiveness. Keys for a successful outcome of this procedures are: thorough preoperative evaluation of patients, experience in open surgery of the oesophagus, and skills in laparoscopy. Based on the literature and on our personal experience, we can conclude that minimally invasive treatment of benign diseases of the oesophagus is safe and effective. In some instances, further follow-up is necessary to confirm the favourable initial results. PMID- 11407677 TI - Medical management of benign oesophageal disease in patients with human immunodeficiency virus infection. AB - Oesophageal diseases are common in human immunodeficiency virus infection, and Candida is the most frequent cause. Empiric therapy with oral antifungal therapy is cost-effective in most patients presenting with oesophageal symptoms. The "gold standard" of diagnosis, endoscopy with brushings and biopsies, is reserved for non-responders within 2 weeks. Since the use of empiric antifungal drugs, the percentage of viral and idiopathic ulcers has increased. The latter frequently recur after treatment, and have been associated with the development of oesophageal strictures. More information is needed on the role of maintenance therapy particularly for viral infections. PMID- 11407678 TI - Diagnostic reliability of immunochemical faecal occult blood test for small colorectal flat adenomas. PMID- 11407679 TI - Ornidazole treatment in normal subjects reduces serum levels of C3. PMID- 11407680 TI - Premature mortality in patients with rheumatoid arthritis: evolving concepts. PMID- 11407681 TI - Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. PMID- 11407682 TI - Time to first occurrence of erosions in inflammatory polyarthritis: results from a prospective community-based study. AB - OBJECTIVE: To examine the time of occurrence of first radiographic erosions in a cohort of patients with inflammatory polyarthritis. METHODS: Patients were recruited through the Norfolk Arthritis Register, which follows up patients annually. Patients with features of rheumatoid arthritis (other than erosions) sufficient, together with erosions, to meet the American College of Rheumatology (formerly, the American Rheumatism Association) 1987 revised criteria were requested to undergo radiographic examinations of the hands and feet at the first and/or second annual followup visits. All patients were requested to undergo radiographic examinations at the fifth annual followup visit. The most recent erosion-free radiograph was identified for 416 eligible patients, and these data were used to derive the duration of disease since the recalled date of onset of first symptoms. The rate of occurrence of first erosions was then determined (as a cumulative prevalence and as an incidence rate using Poisson regression) from analysis of followup films. Patients were assumed to be free of erosions at symptom onset. RESULTS: The cumulative prevalence of erosions in patients whose first film was obtained 12-24 months after disease onset was 36%, equivalent to an incidence rate of 24.5/1,000 patient-months. We identified 3 analysis groups of patients who were free of erosions based on films obtained 12-24 months, 24-36 months, and 36-60 months since the recalled date of onset of first symptoms. New erosions were observed in all 3 groups, with cumulative prevalences of 23%, 28%, and 47%, respectively. These were equivalent to first-erosion incidence rates/1,000 patient-months of 5.4 (95% confidence interval [95% CI] 3.8-83), 6.8 (95% CI 4.7-10.0), and 13.0 (95% CI 8.9-19.2), respectively. CONCLUSION: Many patients with erosive disease first develop their erosions >2 years from disease onset. PMID- 11407683 TI - Change in bone mineral density in patients with rheumatoid arthritis during the first decade of the disease. AB - OBJECTIVE: Rheumatoid arthritis (RA) has been reported to be associated with bone loss during the first years of the disease. The magnitude of this problem after the initial years has not yet been evaluated. In the present study, the change in bone mineral density (BMD) in patients with recent-onset RA as well as the effects of inflammation, mobility, and the use of prednisone on this change were studied in the first decade of the disease. METHODS: BMD was measured twice in 76 RA patients with mean disease durations of 2.35 years at the first BMD measurement and 8.90 years at the second BMD measurement. BMD was measured in both hips using dual x-ray absorptiometry. Results were expressed as mean +/- SEM Z scores (using age- and sex-matched reference values) and as mean +/- SEM percent change in BMD (in gm/cm2) per year. The effects of inflammation, mobility, and the use of prednisone on change in BMD were evaluated using multiple linear regression analyses. RESULTS: At the first BMD measurement, RA patients had lower BMD compared with the reference values (Z score -0.42+/-0.11, 95% confidence interval [95% CI] -0.64, -0.20). Between the 2 measurements, we observed a small decrease in BMD of -0.28+/-0.11%/year (95% CI -0.07 to -0.49). However, the rate of bone loss was smaller than expected. The Z score increased by 0.13+/-0.05 between the 2 BMD measurements (95% CI 0.02, 0.23). Only the use of prednisone was significantly associated with increased bone loss. In a separate analysis that included only postmenopausal women, increased physical activity and longer time since menopause were both associated with decreased bone loss. In this subgroup of patients, the use of prednisone was significantly associated with increased bone loss as well. A high erythrocyte sedimentation rate was associated with increased bone loss, but this did not reach statistical significance. CONCLUSION: After the initial years of the disease, bone loss in RA patients is lower than expected compared with age- and sex-matched reference values. Postmenopausal RA patients with low levels of physical activity are at increased risk of losing bone. Use of prednisone was the only variable consistently associated with reduction in BMD in RA patients. PMID- 11407684 TI - Primary association of a MICA allele with protection against rheumatoid arthritis. AB - OBJECTIVE: To determine whether major histocompatibility complex class I chain related gene A (MICA) polymorphisms are associated with susceptibility to rheumatoid arthritis (RA) independently of the HLA-DRB1 shared epitope (SE). METHODS: Fifty-four Spanish families with an affected son or daughter and 211 consecutive RA patients were genotyped for HLA-DRB1, tumor necrosis factor a/b microsatellite alleles, and MICA transmembrane polymorphism. We performed a case control comparison with the consecutive patients and an independent transmission disequilibrium test with the families. RESULTS: The frequency of the MICA 6.0 allele was significantly reduced, compared with controls, in the group of SE+ patients (odds ratio 0.39, P = 0.0005). Additionally, the haplotypes containing this allele were preferentially not transmitted to the affected offspring (9 transmitted of 33; P = 0.007), independent of the presence or absence of an SE either in the same haplotype or in the other haplotype in the progenitor. CONCLUSION: These data suggest that the MICA 6.0 allele is an independent marker of protection against RA in the SE+ group of RA patients. PMID- 11407685 TI - A novel single-nucleotide polymorphism at the 5'-flanking region of SAA1 associated with risk of type AA amyloidosis secondary to rheumatoid arthritis. AB - OBJECTIVE: To address whether the gamma haplotype at exon 3 of the SAA1 gene is directly associated with type AA amyloidosis or is merely in linkage with an unknown polymorphism that is primarily associated with disease risk, we examined the SAA1 gene for new polymorphisms. METHODS: We analyzed DNA samples from 44 rheumatoid arthritis (RA) patients with AA amyloidosis (amyloid group), 55 RA patients without AA amyloidosis (RA group), and 58 non-RA healthy subjects (non RA group). We also examined DNA samples from 50 Caucasians to compare linkage disequilibrium relationships involving SAA1 region polymorphisms between Japanese and Caucasoid populations. RESULTS: We observed 3 novel single-nucleotide polymorphisms (SNPs) in the 5'-flanking region of SAA1: -61C/G, -13T/C, and 2G/A. Comparison of allele frequencies and ratios of individuals with particular alleles between the study groups revealed statistically significant differences between the amyloid and RA groups and between the amyloid and non-RA groups. Statistical analysis revealed that the -13T/C SNP was strongly associated with AA amyloidosis. In addition, we found tight linkage between the -13T allele and the alpha haplotype, rather than the beta haplotype, at exon 3 in the Caucasoid population, while -13T was closely linked to the gamma and beta haplotypes, rather than the alpha haplotype, in the Japanese population. Since the linkage disequilibrium relationship was reversed between the Japanese and Caucasoid populations, different exon 3 haplotypes of SAA1 are found to be associated with the risk of AA amyloidosis in different ethnic groups. CONCLUSION: Our data suggest that the SAA1 -13T allele, rather than SAA1 exon 3 haplotypes, is primarily associated with AA amyloidosis risk. PMID- 11407686 TI - Regulation of synoviocyte phospholipase A2 and cyclooxygenase 2 by macrophage migration inhibitory factor. AB - OBJECTIVE: Macrophage migration inhibitory factor (MIF) is a proinflammatory cytokine with known actions in macrophage and T cell activation. MIF also has the unique capacity to reverse the inhibitory effects of glucocorticoids on these cells. We have recently demonstrated MIF expression in human rheumatoid arthritis (RA) synovium and cultured fibroblast-like synoviocytes (FLS), as well as the ability of FLS-derived MIF to induce monocyte release of tumor necrosis factor alpha. We investigated the effects of MIF on aspects of RA FLS activation, including the induction of phospholipase A2 (PLA2) and cyclooxygenase (COX). METHODS: PLA2 activity was measured by 3H-arachidonic acid released from treated FLS supernatants. COX activity was measured by prostaglandin E2 enzyme-linked immunosorbent assay. Cytosolic PLA2 (cPLA2) and COX-2 messenger RNA (mRNA) were determined using semiquantitative reverse transcriptase-polymerase chain reaction. RESULTS: Constitutive PLA2 activity was detected in RA FLS. Recombinant human MIF up-regulated PLA2 activity (P < 0.01) and cPLA2 mRNA expression, but had no effect on secretory PLA2. Recombinant human MIF up-regulated COX activity (P < 0.05) and COX-2 mRNA, but had no observable effect on COX-1. Interleukin 1beta (IL-1beta) significantly up-regulated PLA2 activity (P < 0.005) and cPLA2 mRNA expression while anti-MIF monoclonal antibody (mAb) significantly inhibited this IL-1beta-induced PLA2 activity (P < 0.02). Anti-MIF mAb significantly reduced IL-1beta-induced COX activity (P < 0.05) and COX-2 mRNA expression. CONCLUSION: MIF exerts a proinflammatory effect on key aspects of RA FLS activation. That anti-MIF mAb inhibited IL-1beta up-regulation of FLS indicates an additional cofactor role for MIF in IL-1beta-induced FLS activation. These data suggest that MIF antagonism has important therapeutic potential in RA. PMID- 11407687 TI - Specific overexpression of rheumatoid arthritis-associated HLA-DR alleles and presentation of low-affinity peptides. AB - OBJECTIVE: To compare levels of HLA-DR expression in rheumatoid arthritis (RA) patients and healthy controls for whom an ordered expression according to the DR alleles is demonstrated and to test the functional consequences of this expression on peptide presentation. METHODS: Using monoclonal antibodies that recognize different DRB1 alleles, DR molecules were quantitated at the surface of the peripheral blood B cells of 23 RA patients and 17 healthy subjects. The functional consequences of the level of DR surface expression was tested using a universal model of antigen presentation and mutated peptides with variable affinities for the T cell receptor. RESULTS: In healthy subjects, surface HLA-DR molecules were expressed at different levels according to allele (DR53, DR4, and DR11 less than DR1 less than DR7 less than DR15). In RA patients, this hierarchy was not conserved and, furthermore, the density of RA-associated DR4 and DR1 molecules was enhanced in patients compared with the basal density in healthy individuals. We demonstrated that an increased expression of DR molecules at the surface of antigen-presenting cells allowed a noteworthy presentation of low affinity peptides that under normal conditions are not efficient in generating a T cell response at physiologic surface density of the DR molecules. CONCLUSION: Our results suggest that the specific overexpression of RA-associated HLA molecules could be responsible for the presentation of low-affinity autopeptides and therefore the activation of peripheral autoreactive T cells. PMID- 11407688 TI - The combination of tumor necrosis factor alpha blockade with interleukin-1 and interleukin-17 blockade is more effective for controlling synovial inflammation and bone resorption in an ex vivo model. AB - OBJECTIVE: Anti-tumor necrosis factor a (anti-TNFalpha) therapy has shown efficacy in the treatment of rheumatoid arthritis (RA). Since interleukin-1 (IL 1), TNFalpha, and IL-17 have many additive and/or synergistic effects in vitro, we tested whether their combined inhibition by soluble receptors would lead to an enhanced effect on ex vivo models of synovial inflammation and bone destruction. METHODS: RA synovium and bone explants were cultured for 7 days in the presence of 1 microg/ml soluble TNFalpha receptor (STNFR; as in current therapy), type II soluble IL-1 receptor (sIL-1RII), or sIL-17R either alone or in combination. Their effects on the production of IL-6 and the release of C-telopeptide of type I collagen (CTX), a marker of type I collagen destruction, were measured by enzyme-linked immunosorbent assay. RESULTS: In synovium, each soluble receptor alone decreased IL-6 production and CTX release by approximately 35% and approximately 55%, respectively. The combination of all 3 receptors was more effective, inhibiting IL-6 production and collagen degradation by up to 70%. Neither sIL-17R, sIL-1RII, or sTNFR alone had no effect (or an effect of <20% inhibition) on IL-6 production in 18%, 33%, and 22%, respectively, of the samples. In bone, sIL-17R, sIL-1RII, and sTNFR decreased IL-6 production by 23%, 50%, and 37%, respectively, while the combination decreased IL-6 production by 75%. A 50% inhibition of CTX release was obtained with sIL-1RII for 63% of the samples versus 38% of the samples with either sTNFR or sIL-17R. However, the combination of all 3 receptors was not more potent than sIL-1RII alone. CONCLUSION: The inhibitory effect of sTNFR on IL-6 production and collagen degradation in RA synovium and bone was increased in combination with sIL-17R and sIL-1RII. These results support the concept of combination therapy, which may increase the percentage of responding patients as well as the degree of individual patient response. PMID- 11407689 TI - Apoptotic cell death is not a widespread phenomenon in normal aging and osteoarthritis human articular knee cartilage: a study of proliferation, programmed cell death (apoptosis), and viability of chondrocytes in normal and osteoarthritic human knee cartilage. AB - OBJECTIVE: Chondrocytes are crucial for adequate matrix balance and function. Cell proliferation and, recently, extensive apoptotic cell death have been reported in osteoarthritic (OA) cartilage. Apoptotic cell death would be an obvious central factor in the initiation and progression of OA, since there is no potential for replacing articular chondrocytes in the adult. Therefore, we studied the occurrence of apoptotic cell disintegration and cell proliferation in OA and normal articular cartilage obtained from the knees of adult donors of all ages. METHODS: Following immunostaining for cellular proteins as well as staining for nuclear DNA, we performed triple-channel confocal laser scanning microscopy on thick cartilage slices to evaluate lacunar emptying and cell viability. Cell proliferation and apoptotic cell death were evaluated morphologically, by immunodetection of the proliferation-associated Ki-67 antigen, and by the TUNEL reaction. RESULTS: With the exception of the calcified layer, we were not able to detect any major (apoptotic or nonapoptotic) cell disintegration in normal young or aged articular knee cartilage. Single apoptotic cells were detected in OA articular knee cartilage. A significant increase in lacunar emptying was observed in late-stage specimens with higher Mankin scores compared with age-matched normal control cartilage specimens, but not in low-grade lesions. A significant (but lesser) increase in empty lacunae was also observed with age in normal cartilage. Cell proliferation was rarely detected in OA cartilage samples and was not detected at all in normal cartilage samples. CONCLUSION: Our results confirm the findings of previous studies showing that cell proliferation occurs in OA cartilage. They also show that, contrary to previous suggestions, apoptotic cell death is not a widespread phenomenon in aging or OA cartilage. PMID- 11407690 TI - Elevated serum B lymphocyte stimulator levels in patients with systemic immune based rheumatic diseases. AB - OBJECTIVE: To determine whether serum levels of B lymphocyte stimulator (BLyS) are elevated in patients with systemic immune-based rheumatic diseases and correlate with serum Ig levels and/or autoantibody titers. METHODS: Sera from 185 patients with various systemic immune-based rheumatic diseases (95 with systemic lupus erythematosus [SLE], 67 with rheumatoid arthritis [RA], 23 with other diagnoses) were assayed for BLyS and Ig. In 7 patients who required arthrocentesis of a swollen knee, coincident serum and synovial fluid samples were assayed for BLyS. Medical charts were retrospectively reviewed for elevated autoantibody titers and proteinuria within a 1-month period before or after collection of sera for BLyS and Ig determination. Sera concurrently collected from 48 normal healthy subjects served as controls. RESULTS: Serum BLyS levels were elevated in 38 of 185 patients (21%) and correlated significantly with serum IgG levels. Serum BLyS levels did not correlate with the patients' age, sex, race, or medications, but correlated positively with anti-double-stranded DNA antibody titers among SLE patients and with rheumatoid factor titers among seropositive RA patients. In contrast, serum BLyS levels correlated inversely with nephrotic-range proteinuria among SLE patients. In every case tested, BLyS levels in clinically inflamed synovial fluids were greater than those in simultaneously obtained sera. CONCLUSION: BLyS may be an important factor in driving polyclonal hypergammaglobulinemia and elevated autoantibody titers in patients with systemic immune-based rheumatic diseases. Local production of BLyS in the joints may contribute to joint pathology. Patients with elevated serum BLyS levels may be ideal candidates for therapeutic targeting of BLyS. PMID- 11407691 TI - Plasma levels of nucleosomes and nucleosome-autoantibody complexes in murine lupus: effects of disease progression and lipopolyssacharide administration. AB - OBJECTIVE: To evaluate the effect of disease progression and lipopolysaccharide (LPS) administration on the presence of nucleosomes, antinucleosome reactivity, and nucleosome-Ig complexes in the circulation of MRL and control mice. METHODS: Plasma samples from lupus-prone (MRL/lpr and MRL/+) and control (CBA, Swiss, and BALB/c) mice were tested in enzyme-linked immunosorbent assays for the presence of nucleosomes, antinucleosome antibodies, and nucleosome-Ig complexes. Nucleosome kinetics, apoptosis induction, and phagocytosis of apoptotic cells were also analyzed in MRL/lpr, MRL/+, and CBA control mice after a single injection of LPS or phosphate buffered saline. RESULTS: Nucleosomes were found in the circulation of MRL/lpr and MRL/+ mice from week 4 onward. Nucleosomes were also detected in young control mice, but with increasing age, the nucleosomes disappeared. Antinucleosome antibodies, nucleosome-Ig complexes, and albuminuria were found only in the MRL/lpr mice. LPS administration led to a significant increase in circulating nucleosomes (3-8-fold) in all strains tested. In only the MRL/lpr mice was this increase followed by a significant decrease in antinucleosome titers and an increase in nucleosome-Ig complexes. The number of apoptotic cells in the thymus after LPS was significantly higher in the MRL/lpr mice than in the MRL/+ and CBA control mice. LPS caused a profound reduction (50 70%) of the phagocytosis of apoptotic cells by peritoneal macrophages, which was comparable for all strains. CONCLUSION: In MRL lupus-prone mice, nucleosomes are persistently present in the circulation, whereas in control mice, nucleosomes are present only at a young age. The formation of antinucleosome antibodies and nucleosome-Ig complexes is a characteristic feature of MRL/lpr mice. LPS administration increases systemic nucleosome release due to an enhancement of apoptosis and a decrease in the clearance of apoptotic cells. PMID- 11407692 TI - Molecular heterogeneity of prolactin in the plasma of patients with systemic lupus erythematosus. AB - OBJECTIVE: Systemic lupus erythematosus (SLE) has been associated with high levels of prolactin in the circulation of some patients. Although prolactin stimulates immune responses, the relationship between hyperprolactinemia and the pathophysiology of SLE remains controversial. This study was undertaken to investigate whether circulating bioactive prolactin isoforms are associated with the activity of SLE. METHODS: The molecular heterogeneity of prolactin was studied in the plasma of patients with active and inactive SLE and in healthy volunteers by radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), Nb2-cell bioassay, and immunoprecipitation-Western blots. The specificity of the bioassay determinations was assessed by neutralization of growth-promoting effects with antiserum to human prolactin. RESULTS: Significantly higher prolactin levels were detected by bioassay and by ELISA than by RIA in both subsets of SLE patients and in normal individuals. Plasma prolactin levels in the SLE patients were significantly greater than those in the normal controls when measured by ELISA, but not by RIA or bioassay. The bioassay:ELISA and bioassay:RIA ratios were similar between SLE patients and controls, suggesting that prolactin biopotency was not altered with the disease, and none of the 3 assays detected a difference in prolactin levels between patients with active SLE and those with inactive SLE. However, the prolactin detected in plasma was associated with immunoreactive proteins of 130 kd and 23 kd, and the concentration of the 130-kd prolactin-like species was 10-fold higher in inactive SLE versus active SLE patients. CONCLUSION: Discrepancies among assays substantiate the molecular heterogeneity of circulating prolactin. The prolactin isotype that is found in association with inactive SLE could be of potential use as a marker for the inactive form of the disease and as an index for the efficacy of treatment. PMID- 11407693 TI - T cell signaling abnormalities in systemic lupus erythematosus are associated with increased mutations/polymorphisms and splice variants of T cell receptor zeta chain messenger RNA. AB - OBJECTIVE: T cells from patients with systemic lupus erythematosus (SLE) display antigen receptor-mediated signaling aberrations associated with defective T cell receptor (TCR) zeta chain protein and messenger RNA (mRNA) expression. This study was undertaken to explore the possibility that coding-region mutations/polymorphisms of the TCR zeta chain could account for its decreased expression and altered signaling in SLE T cells. METHODS: TCR zeta chain mRNA from 48 SLE patients, 18 disease controls, and 21 healthy volunteers was reverse transcribed, amplified by polymerase chain reaction, and cloned, and complementary DNA (cDNA) was sequenced. DNA sequences from multiple clones were analyzed for silent single-nucleotide polymorphisms, mutations, and splice variations, to promote the identification of heterozygosity. RESULTS: DNA sequence analysis revealed several widely distributed missense mutations and silent polymorphisms in the coding region of the TCR zeta chain, which were more frequent in SLE patients than in patients with other rheumatic diseases or healthy controls (P < 0.0001). Several of the missense mutations were located in the 3 immunoreceptor tyrosine activation motifs or the GTP binding domain, and this could lead to functional alterations in the TCR zeta chain. A splice variant of the TCR zeta chain with a codon CAG (glutamine) insertion between exons IV and V was found in half of the SLE and control samples. Two larger spliced isoforms of the TCR zeta chain, with an insertion of 145 bases and 93 bases between exons I and II, were found only in SLE T cells. We also identified various alternatively spliced forms of the TCR zeta chain resulting from the deletion of individual exons II, VI, or VII, or a combined deletion of exons V and VI; VI and VII; II, III, and IV; or V, VI, and VII in SLE T cells. The frequency of the deletion splice variants was significantly higher in SLE than in control samples (P = 0.004). These variations were observed in cDNA and may not reflect the status of the genomic DNA. CONCLUSION: These findings demonstrate that heterogeneous mutations/polymorphisms and alternative splicing of TCR zeta chain cDNA are more frequent in SLE T cells than in T cells from non-SLE subjects and may underlie the molecular basis of known T cell signaling abnormalities in this disease. PMID- 11407694 TI - A randomized, controlled trial of methotrexate versus placebo in early diffuse scleroderma. AB - OBJECTIVE: Early diffuse scleroderma (systemic sclerosis; SSc) has no proven treatment. This study was undertaken to examine the efficacy of methotrexate (MTX) in improving the skin and other disease parameters in early diffuse SSc. METHODS: Seventy-one patients with diffuse SSc of <3 years' duration were enrolled in a multicenter, randomized, placebo-controlled, double-blind trial. Thirty-five patients were treated with MTX and 36 with placebo. Treatment was administered for 12 months. The primary outcome measures were skin score (as determined with 2 different indices) and physician global assessment. RESULTS: At baseline, there were no statistically significant differences in skin scores, carbon monoxide diffusing capacity (DLco), physician global assessment, or other secondary outcome measurements between the 2 treatment groups. At study completion, results slightly favored the MTX group (mean +/- SEM modified Rodnan skin score 21.4+/-2.8 in the MTX group versus 26.3+/-2.1 in the placebo group [P < 0.17]; UCLA skin score 8.8+/-1.2 in the MTX group versus 11.0+/-0.9 in the placebo group [P < 0.15]; DLco in the MTX group 75.7+/-4.6 versus 61.8+/-3.4 in the placebo group [P < 0.2]). In addition, physician global assessment results favored MTX (P < 0.035), whereas patient global assessment did not differ significantly between groups. When between-group differences for changes in scores from baseline to 12 months were examined using intent-to-treat methodology, MTX appeared to have a favorable effect on skin scores (modified Rodnan score -4.3 in the MTX group versus 1.8 in the placebo group [P < 0.009]; UCLA score -1.2 in the MTX group versus 1.2 in the placebo group [P < 0.02]), but differences in the degree of change in the DLco and physician global assessment were not significant. For the UCLA skin score, these differences in results were not statistically significant after adjustment for baseline differences in sex distribution and steroid use. Dropout rates were similar in the 2 groups. CONCLUSION: Although results of this trial demonstrated a trend in favor of MTX versus placebo in the treatment of early diffuse SSc, the between-group differences were small and the power to rule out false-negative results was only 50%. Our findings do not provide evidence that MTX is significantly effective in the treatment of early diffuse SSc. PMID- 11407695 TI - Familial occurrence frequencies and relative risks for systemic sclerosis (scleroderma) in three United States cohorts. AB - OBJECTIVE: To determine the frequency with which scleroderma (systemic sclerosis; SSc) recurs in families and the familial relative risk (lambda) in the US. METHODS: Family histories of SSc were prospectively surveyed in 3 large US cohorts of SSc patients, 2 in Texas and 1 in Michigan. Diagnoses of familial SSc were verified by rheumatologist evaluation and/or review of medical records. Familial relative risks for first-degree relatives (lambda1) and siblings (lambdas) were calculated using actual reported counts of first-degree relatives in 2 cohorts and recent estimates of SSc prevalence in the US. RESULTS: Compared with the estimated prevalence of SSc in the US (2.6 cases/10,000 population [0.026%]), the disease occurred in 1 or more first-degree relatives in 1.5-1.7% of SSc families in the 3 cohorts (or 11 of 703 families [1.6%]), a significant increase. Familial relative risks in first-degree relatives in the 3 cohorts ranged from 10 to 16 (13 combined), and in siblings they ranged from 10 to 27 (15 combined). CONCLUSION: SSc occurs significantly more frequently in families with scleroderma (1.6%) than in the general population (0.026%). A positive family history of SSc is the strongest risk factor yet identified for SSc; however, the absolute risk for each family member remains quite low (<1%). PMID- 11407696 TI - Clinical significance of surfactant protein D as a serum marker for evaluating pulmonary fibrosis in patients with systemic sclerosis. AB - OBJECTIVE: To determine the clinical significance of surfactant protein D (SP-D), a useful marker for evaluating various lung diseases, in patients with systemic sclerosis (SSc) and to clarify any clinical significance between SP-D and KL-6, which is known to be correlated with pulmonary fibrosis (PF) in SSc patients. METHODS: We used a specific enzyme-linked immunosorbent assay to measure serum SP D levels in 83 patients with SSc and 31 healthy control subjects. RESULTS: The serum levels of SP-D were significantly higher in patients with SSc than in healthy controls (mean +/- SD 81.9+/-59.2 versus 34.8+/-13.7 ng/ml). Serum SP-D levels in patients with diffuse cutaneous SSc were significantly higher than those in patients with limited cutaneous SSc (98.8+/-72.1 versus 66.8+/-40.0 ng/ml). Serum SP-D levels in patients with PF were significantly elevated compared with those in patients without PF (99.7+/-64.1 versus 65.3+/-49.4 ng/ml). Moreover, the incidences of decreased percentage diffusing capacity for carbon monoxide and decreased percentage vital capacity were also significantly greater in patients with elevated SP-D levels than in those with normal levels (67% versus 43% and 36% versus 17%, respectively). There was a significant positive correlation between serum levels of SP-D and KL-6. Serum SP-D and KL-6 levels showed almost the same sensitivities and specificities in the diagnosis of PF (68% versus 73% and 70% versus 74%, respectively). These two markers also predicted PF to almost the same degree (31% versus 33%, respectively). CONCLUSION: These results suggest that SP-D, as well as KL-6, may be a useful serum marker for evaluating PF in patients with SSc. PMID- 11407697 TI - Depletion of protein kinase Cepsilon in normal and scleroderma lung fibroblasts has opposite effects on tenascin expression. AB - OBJECTIVE: To determine whether the extracellular matrix protein tenascin-C (TN C) is overexpressed in lung fibroblasts from systemic sclerosis (SSc) patients, the molecular mechanisms regulating TN-C secretion in SSc and normal lung fibroblasts, and how these processes might contribute to lung fibrosis in SSc patients. METHODS: TN-C secretion by SSc and normal fibroblasts was compared in vivo (in bronchoalveolar lavage [BAL] fluid) and in vitro (in culture medium). The ability of thrombin to induce TN-C was confirmed at both the protein and the messenger RNA (mRNA) level. The role of protein kinase Cepsilon (PKCepsilon) in the expression of TN-C was evaluated by determining the effects of thrombin on PKCepsilon levels and by directly manipulating PKCepsilon levels via the use of antisense oligonucleotides. RESULTS: BAL fluid from SSc patients contained high levels of TN-C, whereas that from normal subjects contained little or no TN-C. In vitro, SSc lung fibroblasts expressed much higher amounts of TN-C than did normal lung fibroblasts. Consistent with the idea that thrombin is a physiologic inducer of TN-C, thrombin stimulated TN-C mRNA and protein expression in both SSc and normal lung fibroblasts by a mechanism that required proteolytic cleavage of the thrombin receptor. Surprisingly, thrombin treatment and antisense oligonucleotide mediated depletion of PKCepsilon indicated that TN-C expression is regulated via opposite signaling mechanisms in SSc and normal cells. In SSc lung fibroblasts, thrombin decreased PKCepsilon levels, and the decreased PKCepsilon induced TN-C secretion; in normal fibroblasts, thrombin increased PKCepsilon levels, and the increased PKCepsilon induced TN-C secretion. Normal and SSc lung fibroblasts also differed in the subcellular localization of PKCepsilon, both before and after thrombin treatment. CONCLUSION: These studies are the first to demonstrate that thrombin is a potent simulator of TN-C in lung fibroblasts and that PKCepsilon is a critical regulator of TN-C protein levels in these cells. Furthermore, our results indicate that both the regulation of PKCepsilon levels by thrombin and the regulation of TN-C levels by PKCepsilon are defective in SSc lung fibroblasts. PMID- 11407698 TI - Chemokine expression by systemic sclerosis fibroblasts: abnormal regulation of monocyte chemoattractant protein 1 expression. AB - OBJECTIVE: Chemokines are important mediators in the chemoattraction of leukocytes to sites of inflammation. This study investigated the potential contribution of systemic sclerosis (SSc) fibroblasts to chemokine production and its potential relevance to the pathogenesis of SSc. METHODS: The expression of messenger RNA (mRNA) for different C-C and C-X-C chemokines by SSc and normal fibroblasts was studied by RNase protection assay. Monocyte chemoattractant protein 1 (MCP-1) protein production was analyzed by enzyme-linked immunosorbent assay. The chemotactic effect of fibroblast-derived MCP-1 on monocytic cells was analyzed in a transmigration assay. Nuclear factor kappaB (NF-kappaB) and activator protein 1 (AP-1) activation in fibroblasts was studied by electromobility shift analysis. MCP-1 expression in SSc skin sections was studied by immunohistochemistry. RESULTS: Among all chemokine genes studied, only MCP-1 and interleukin-8 mRNA were expressed by nonstimulated normal and SSc fibroblasts. SSc fibroblasts displayed increased constitutive expression of MCP-1 mRNA and protein and showed a blunted response to oxidative stress. Increased MCP 1 production was associated with higher chemotactic activity for monocytic cells. Increased NF-kappaB or AP-1 activation was not responsible for the constitutive overexpression of MCP-1 by SSc fibroblasts. In SSc skin sections, MCP-1 expression was detected in fibroblasts, keratinocytes, and mononuclear cells, whereas it was undetectable in normal skin. CONCLUSION: SSc fibroblasts display a specific pattern of chemokine gene expression that is characterized by constitutively increased and abnormally regulated expression of MCP-1 in vitro. MCP-1 is also expressed in lesional skin and can participate in the pathogenesis of SSc. PMID- 11407699 TI - Small-vessel vasculitis surrounding a spared temporal artery: clinical and pathological findings in a series of twenty-eight patients. AB - OBJECTIVE: Occasionally, a temporal artery biopsy reveals small-vessel vasculitis (SVV) surrounding a spared temporal artery, the significance of which is unclear. We analyzed the final diagnosis in a series of patients with this condition and tried to identify histopathologic features with potential usefulness in predicting the ultimate diagnosis. METHODS: We performed a clinical and histopathologic review of 28 patients in whom SVV surrounding a spared temporal artery was the first histologic finding that led to the diagnosis of vasculitis. For comparison purposes, we analyzed the pattern of small vessel involvement in 30 patients with biopsy-proven giant cell arteritis (GCA). RESULTS: GCA was considered the most likely diagnosis in 12 patients, based on the absence of clinical evidence of additional organ involvement and normal findings on muscle biopsy and electrophysiologic study. Three patients had systemic necrotizing vasculitis (SNV), based on the demonstration of typical lesions on subsequent muscle, nerve, or kidney biopsy. After extensive evaluation, 4 patients remained unclassifiable. Nine patients were incompletely studied. Fibrinoid necrosis was significantly more frequent in patients with SNV (P = 0.0022), whereas involvement of vasa vasorum was more frequent in patients classified as having GCA (P = 0.022). No differences in the pattern of small vessel involvement were found in patients with SVV surrounding a spared temporal artery who were classified as having GCA compared with patients with biopsy-proven GCA. Granulocytes were observed at similar frequency in all conditions. CONCLUSION: SVV may be the only abnormal feature in a temporal artery biopsy and the only histologic evidence of vasculitis. The diagnosis of GCA can be reasonably established in most of these patients when there is no apparent evidence of additional organ involvement. However, when fibrinoid necrosis is observed or the temporal artery vasa vasorum are not involved, SNV must be extensively excluded. PMID- 11407700 TI - Is disease severity in ankylosing spondylitis genetically determined? AB - OBJECTIVE: To assess the role of genes and the environment in determining the severity of ankylosing spondylitis. METHODS: One hundred seventy-three families with >1 case of ankylosing spondylitis were recruited (120 affected sibling pairs, 26 affected parent-child pairs, 20 families with both first- and second degree relatives affected, and 7 families with only second-degree relatives affected), comprising a total of 384 affected individuals. Disease severity was assessed by the Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) and functional impairment was determined using the Bath Ankylosing Spondylitis Functional Index (BASFI). Disease duration and age at onset were also studied. Variance-components modeling was used to determine the genetic and environmental components contributing to familiality of the traits examined, and complex segregation analysis was performed to assess different disease models. RESULTS: Both the disease activity and functional capacity as assessed by the BASDAI and the BASFI, respectively, were found to be highly familial (BASDAI familiality 0.51 [P = 10(-4)], BASFI familiality 0.68 [P = 3 x 10(-7)]). No significant shared environmental component was demonstrated to be associated with either the BASDAI or the BASFI. Including age at disease onset and duration of disease as covariates made no difference in the heritability assessments. A strong correlation was noted between the BASDAI and the BASFI (genetic correlation 0.9), suggesting the presence of shared determinants of these 2 measures. However, there was significant residual heritability for each measure independent of the other (BASFI residual heritability 0.48, BASDAI 0.36), perhaps indicating that not all genes influencing disease activity influence chronicity. No significant heritability of age at disease onset was found (heritability 0.18; P = 0.2). Segregation studies suggested the presence of a single major gene influencing the BASDAI and the BASFI. CONCLUSION: This study demonstrates a major genetic contribution to disease severity in ankylosing spondylitis. As with susceptibility to ankylosing spondylitis, shared environmental factors play little role in determining the disease severity. PMID- 11407701 TI - Host metalloproteinases in Lyme arthritis. AB - OBJECTIVE: To assess the role of matrix metalloproteinases (MMPs) in cartilage and bone erosions in Lyme arthritis METHODS: We examined synovial fluid from 10 patients with Lyme arthritis for the presence of MMP-2, MMP-3, MMP-9, and "aggrecanase" activity using gelatinolytic zymography and immunoblot analysis. We developed an in vitro model of Lyme arthritis using cartilage explants and observed changes in cartilage degradation in the presence of Borrelia burgdorferi and/or various protease inhibitors. RESULTS: Synovial fluid from patients with Lyme arthritis was found to contain at least 3 MMPs: gelatinase A (MMP-2), stromelysin (MMP-3), and gelatinase B (MMP-9). In addition, there was evidence in 2 patients of "aggrecanase" activity not accounted for by the above enzymes. Infection of cartilage explants with B. burgdorferi resulted in induction of MMP 3, MMP-9, and "aggrecanase" activity. Increased induction of these enzymes by B. burgdorferi alone was not sufficient to cause cartilage destruction in the explants as measured by glycosaminoglycan (GAG) and hydroxyproline release. However, addition of plasminogen, which can act as an MMP activator, to cultures resulted in significant GAG and hydroxyproline release in the presence of B. burgdorferi. The MMP inhibitor batimastat significantly reduced the GAG release and completely inhibited the collagen degradation. CONCLUSION: MMPs are found in synovial fluids from patients with Lyme arthritis and are induced from cartilage tissue by the presence of B. burgdorferi. Inhibition of MMP activity prevents B. burgdorferi-induced cartilage degradation in vitro. PMID- 11407702 TI - Etanercept therapy in children with treatment-resistant uveitis. AB - OBJECTIVE: To evaluate the safety and efficacy of the tumor necrosis factor fusion protein etanercept in children with treatment-resistant uveitis. METHODS: Ten children with chronic active uveitis (7 girls and 3 boys, mean age 7.5 years [range 3-12 years]) were enrolled in this prospective study. In 7 children, uveitis was associated with pauciarticular juvenile rheumatoid arthritis. Five children were antinuclear antibody positive. All patients had failed previous therapy with topical steroids and methotrexate and/or cyclosporine. All were treated with etanercept at a dosage of 0.4 mg/kg twice weekly for the first 3 months, and then, if eyes did not improve, with 25 mg twice weekly (mean 1.1 mg/kg) for at least 3 additional months. RESULTS: At the beginning of the trial, uveitis affected 18 eyes in the 10 children. Within 3 months, 10 of 16 affected eyes (63%; P = 0.017) showed a rapid decrease in anterior chamber cell density, including remission of uveitis in 4 eyes. In children with visual acuity of less than 20/25, 4 of 10 eyes (40%) improved. An exacerbation of uveitis during etanercept therapy occurred in only 1 child (1 of 14 eyes [7%]). Other ocular outcome parameters, such as intraocular pressure, synechia formation, and lens clarity, remained unchanged. Following a dosage increase to an average of 1.1 mg/kg after 3 months in 7 children, no further improvement was noted. CONCLUSION: Our data suggest that etanercept injected subcutaneously twice a week has a beneficial effect on treatment-resistant chronic uveitis in children. Further controlled studies with etanercept in systemic or topical form are necessary to confirm its efficacy and optimal mode of administration. PMID- 11407703 TI - The musculoskeletal manifestations of familial Mediterranean fever in children genetically diagnosed with the disease. AB - OBJECTIVE: Familial Mediterranean fever (FMF) is characterized by recurrent episodes of peritonitis, pleuritis, and synovitis. Its most common musculoskeletal manifestation is acute recurrent monarthritis, but other manifestations have also been described. We describe the articular and musculoskeletal manifestations in a group of patients who were found by genetic screening to be homozygous for the FMF gene. METHODS: We surveyed 136 pediatric patients of Mediterranean extraction who were evaluated for a variety of musculoskeletal symptoms, and in whom genetic studies confirmed a diagnosis of FMF. Two groups of patients emerged: group 1 contained 107 patients who displayed a classic picture of FMF, and group 2 comprised 29 patients whose symptoms did not fulfill the criteria for a clinical diagnosis of FMF. Fifty-nine patients were Sephardic Jews and 77 were Arabs. The Jewish patients were all homozygous or compound heterozygous for the M694V mutation, while the Arab patients were homozygous or compound heterozygous for any 1 of the 5 mutations tested (M694V, V726A, M680I, M694I, and E148Q). RESULTS: Acute episodes of monarthritis occurred in 42 (71%) of the Jewish children and 31 (40%) of the Arab children; 70% of these patients had the M694V mutation. Acute monarthritis occurred in 73 (68%) of the patients of group 1, but in none of the patients from group 2. Ten (34%) of the 29 patients from group 2 exhibited diverse musculoskeletal manifestations. Thirteen patients in our series (10%) presented with a variety of musculoskeletal symptoms, including febrile myalgia syndrome in 6 patients. CONCLUSION: Acute episodes of monarthritis are the most common musculoskeletal manifestation of FMF in children bearing the M964V mutation, which predominates among Sephardic Jews, although children with the M694V mutation may also present with diverse nonspecific musculoskeletal manifestations. Genetic screening for FMF appears indicated in the evaluation of unexplained musculoskeletal symptoms in children of Mediterranean extraction. PMID- 11407704 TI - Exacerbation of antigen-induced arthritis in inducible nitric oxide synthase deficient mice. AB - OBJECTIVE: Inhibition of nitric oxide (NO) produced by inducible NO synthase (iNOS) is suggested to be beneficial in experimental arthritis. Although NO is important for the integrity of the microcirculation, the effects of inhibition of iNOS on the synovial microcirculation are not currently known. This study investigated the synovial microcirculation and leukocyte-endothelial cell interactions in iNOS-deficient mice with antigen-induced arthritis (AIA) and compared these findings with disease severity. METHODS: Fourteen homozygous iNOS /- and 14 iNOS+/+ mice were used. The severity of AIA was assessed by measuring knee joint swelling and by histologic scoring. The number of rolling and adherent leukocytes was quantitatively analyzed in synovial microvessels using intravital microscopy of intraarticular synovial tissue. Nitrite/nitrate concentrations were measured, and the expression of iNOS, E- and P-selectin, intercellular adhesion molecule 1, and vascular cell adhesion molecule 1 (VCAM-1) was assessed by immunohistochemistry. RESULTS: In iNOS+/+ animals with AIA, the plasma concentration of nitrite/nitrate was increased 3-fold and iNOS expression was detected in cells of the joint. Swelling of the knee joint as well as leukocyte infiltration were enhanced in the iNOS-/- arthritic animals compared with iNOS+/+ mice with AIA. AIA-associated leukocyte-endothelial cell interaction in synovial postcapillary venules was more pronounced in iNOS-/-, compared with iNOS+/+, arthritic mice. A strong expression of P-selectin and VCAM-1 was observed in the iNOS-/- arthritic mice only. CONCLUSION: These data suggest that NO production by iNOS in vivo has antiinflammatory effects in experimental arthritis, by mediating a reduction in leukocyte adhesion and infiltration. PMID- 11407705 TI - E-selectin, but not P-selectin, is required for development of adjuvant-induced arthritis in the rat. AB - OBJECTIVE: To determine the role of the endothelial cell adhesion molecules E- and P-selectin in the development and severity of adjuvant-induced arthritis (AIA) in the rat. METHODS: Lewis rats were immunized subcutaneously with Mycobacterium butyricum (Mb), and blocking monoclonal antibodies (mAb) to rat E- and P-selectin were administered. Clinical score, radiolabeled (51Cr and 111In) blood polymorphonuclear leukocyte (PMN) and monocyte migration to joints, and histologic features were monitored. RESULTS: When mAb treatment was started on day 5 postimmunization with Mb (preclinical stage), development of AIA was significantly (P < 0.01) inhibited by mAb to E- but not to P-selectin (mean score on day 14 control 10.2, anti-E 2.8, anti-P 9.1). This was associated with markedly decreased migration (by 66-94%) of PMN and monocytes to arthritic joints and diminished cartilage degradation. When treatment was delayed until animals showed signs of arthritis (day 10 postimmunization), only a marginal and variable effect was observed as compared with blockade during the preclinical (day 5) stage. E-selectin blockade on day 5 and day 7 postimmunization resulted in inhibition of antigen-dependent T cell-mediated inflammation, since it decreased T cell migration to sites of dermal-delayed hypersensitivity induced by Mb without affecting migration to concanavalin A or cytokines. The proliferative response of T cells to Mb in vitro was not altered. CONCLUSION: E-selectin plays an important role early in the development of AIA. This adhesion molecule may contribute to the migration of antigen-reactive T cells to peripheral tissues, including the joints where T cells initiate the arthritis. PMID- 11407706 TI - Germinal center reaction in the joints of mice with collagen-induced arthritis: an animal model of lymphocyte activation and differentiation in arthritis joints. AB - OBJECTIVE: To establish an animal model and provide a basis for investigating the role of germinal center (GC) reaction in autoimmune arthritis. METHODS: DBA/1 mice were immunized with bovine type II collagen (CII) to elicit collagen-induced arthritis (CIA). Sections of arthritic joints were examined by in situ immunohistochemical studies, and purified cells from affected joints were subjected to flow cytometry analysis. RESULTS: De novo GC reaction was induced in the arthritic joints of male DBA/1 mice by immunization with bovine CII. In comparison with GCs formed in lymphoid tissues, such as spleen and lymph nodes, we found that these GCs formed in the joint tissues of mice with CIA were morphologically typical, as determined by immunohistologic and flow cytometric assays. CONCLUSION: The local immune responses in murine CIA induced ectopic GC formation, as observed in the synovial tissues of patients with rheumatoid arthritis. This system will allow for the first time the direct study of the role of the GC reaction in autoimmune arthritis in an animal model. PMID- 11407707 TI - Paradoxical effects of tissue inhibitor of metalloproteinases 1 gene transfer in collagen-induced arthritis. AB - OBJECTIVE: The imbalance between matrix metalloproteinases (MMPs) 1, 3, and 9 and their specific inhibitor, tissue inhibitor of metalloproteinases 1 (TIMP-1), is a critical step in cartilage injury and angiogenesis in arthritis. To explore the therapeutic potential of TIMP-1 gene transfer in erosive arthritis, the effects of an adenoviral vector (Ad-TIMP-1) were assessed in DBA/1 mice with collagen induced arthritis (CIA). METHODS: DBA/1 mice with CIA received an intravenous injection of replication-deficient adenovirus containing the human TIMP-1 gene or a control LacZ gene on day 28 postimmunization. The efficiency of gene transfer was determined by serum TIMP-1 detection, measurements of paw swelling, as well as radiologic and histologic examination of the paws. RESULTS: A single administration of Ad-TIMP-1 resulted in detectable serum levels of the exogenous protein for at least 13 days. The incidence and onset of arthritis were not statistically modified after human TIMP-1 gene transfer in DBA/1 mice compared with control mice. However, the severity of inflammation was statistically significantly increased in Ad-TIMP-1-treated mice and a similar trend was observed in the histologic and radiologic scores. With regard to the mechanisms of the worsened effect in the Ad-TIMP-1-treated mice, we observed 1) higher serum levels of anti-type II collagen IgG2a, 2) a significant increase in endogenous soluble tumor necrosis factor receptor I (TNFRI) in sera, and 3) increased labeling of mouse tumor necrosis factor alpha and TNFRI within arthritic joints. CONCLUSION: These findings show that overexpression of TIMP-1 does not prevent osteochondral injury in a mouse model of arthritis. Since MMPs have overlapping properties in terms of their roles in extracellular matrix degradation, angiogenesis, and shedding of cell surface adhesion molecules, cytokines, and cytokine receptors, the paradoxical results obtained suggest that TIMP-1 is probably not the main inhibitor to target. PMID- 11407708 TI - Matrix metalloproteinases and aggrecanases cleave aggrecan in different zones of normal cartilage but colocalize in the development of osteoarthritic lesions in STR/ort mice. AB - OBJECTIVE: To map aggrecan cleavage by matrix metalloproteinases (MMPs) and aggrecanases in normal murine tibial articular cartilage (CBA strain) and in the development of spontaneous osteoarthritis (OA) in the STR/ort mouse and to assess the influence of sex hormone status on these conditions in gonadectomized STR/ort mice. METHODS: The distributions of neoepitopes of aggrecan generated by MMP (VDIPEN) and aggrecanase (NITEGE) cleavage were investigated by immunohistochemistry. RESULTS: VDIPEN neoepitope was detected mainly in the pericellular matrix of deep-zone chondrocytes in normal tibial cartilage from STR/ort and CBA mice. In early OA, VDIPEN immunostaining also localized to the pericellular matrix of chondrocytes at the site of the lesion. With increasing severity of OA lesions, VDIPEN immunostaining was also detected in the interterritorial matrix, close to the site of the lesion. In contrast, NITEGE mapped most strongly to the pericellular matrix of upper-zone chondrocytes in normal tibial cartilage. As with VDIPEN, NITEGE was strongly expressed in the pericellular matrix at the site of early OA lesions. With advancing OA, NITEGE colocalized with VDIPEN in both the pericellular and interterritorial matrices of chondrocytes adjacent to OA lesions and in those of the deep zones. Hormone status did not appear to influence the development of OA or the distribution of aggrecan neoepitopes in STR/ort mice. CONCLUSION: MMP- and aggrecanase-generated neoepitopes map predominantly to different regions in normal murine tibial cartilage. However, both groups of enzymes generate increased amounts of neoepitopes in pericellular and interterritorial matrix adjacent to histopathologic lesions of OA. Aggrecan degradation and the development of OA appear to be independent of sex hormone status in this model. PMID- 11407709 TI - High thrombosis rate after fetal loss in antiphospholipid syndrome: effective prophylaxis with aspirin. PMID- 11407710 TI - Recent improvements in survival in patients with rheumatoid arthritis: better outcomes or different study designs? PMID- 11407711 TI - Treatment of intraarticular osteoarthritis of the knee with hylan G-F 20: comment on the article by Brandt et al. PMID- 11407712 TI - Efficacy of intraarticular hyaluronan in the treatment of knee osteoarthritis: comment on the article by Brandt et al. PMID- 11407713 TI - Ankylosing spondylitis, birth order, and hormones: comment on the article by Baudoin et al. PMID- 11407714 TI - MRI of acute spinal trauma. AB - The clinician managing patients who have suffered trauma to the spine requires several questions answered from imaging studies. In the acute stage, a full assessment of the complete injury to the bony, ligamentous, disc and neural tissues will determine the stability of the injury and help decide the nature of clinical management, either conservative or surgical, and also help in determining the surgical approach. Magnetic resonance imaging (MRI) is established as a vital imaging technique and can answer many of the questions posed above. The purpose of this article is to review the current status of MRI in the assessment of acute spinal trauma. PMID- 11407715 TI - Subchondral insufficiency fracture of the femoral head: histopathologic correlation with MRI. AB - OBJECTIVE: To correlate the magnetic resonance imaging (MRI) features with the histopathologic findings in subchondral insufficiency fracture (SIF) of the femoral head. DESIGN AND PATIENTS: This study was based on a retrospective review of the MRI features and histopathologic findings in seven patients with SIF who had had total hip replacement. Results. In all seven cases, MRI showed a bone marrow edema pattern in the femoral head, and a focal low-intensity band beneath the articular cartilage on some slices (not all) on the T1-weighted images. The shape of the low-intensity band varied: it was irregular and serpentine in four cases, well-delineated, smooth, and a mirror image to the articular surface in two cases, and parallel to the articular surface in one case. On histologic examination, the low-intensity band on MRI corresponded to a fracture line and its associated repair tissue. In all but one case, the band was not visible on T2 weighted or fat suppression images, and the proximal subchondral portion of the lesion had a homogeneous high signal intensity. This region of high signal intensity corresponded histopathologically to viable bone and marrow tissue with associated callus, edema, and vascular granulation tissue. CONCLUSIONS: SIF of the femoral head characteristically demonstrates a low-intensity band on T1 weighted images that corresponds, histopathologically, to a linear subchondral fracture and its associated repair tissue. In most cases, the subchondral portion of the lesion appears on T2-weighted images as an area of homogeneously high signal intensity. PMID- 11407716 TI - Subchondral changes in transient osteoporosis of the hip. AB - OBJECTIVE: To review the subchondral changes on MR imaging in transient osteoporosis of the hip (TOH) and to consider the pathophysiology. DESIGN AND PATIENTS: MR images of 12 hips of 11 consecutive patients with TOH were retrospectively studied. The diagnoses of TOH were confirmed on the basis of previously published criteria, including decreased bone density of the femoral head and/or neck on radiographs, bone marrow edema (BME) pattern on MR images, spontaneous resolution of the symptoms and a return to normal radiodensity. RESULTS: All 12 hips showed a BME pattern in the femoral head and/or neck. Linear patterns of very low signal intensity were identified on T1-weighted images in the subchondral area within the diffuse low signal intensity area in all 12 hips. On T2-weighted images, a low signal intensity line was observed in the corresponding area in eight hips only. These linear patterns were thought to represent subchondral fracture lines. CONCLUSIONS: The presence of a subchondral fracture may be important when considering the pathophysiology of TOH. PMID- 11407717 TI - Angled oblique sagittal MR imaging of rotator cuff tears: comparison with standard oblique sagittal images. AB - OBJECTIVE: To compare the accuracy for diagnosing rotator cuff tears of oblique coronal images supplemented with standard oblique sagittal images versus thinner section angled oblique sagittal images. DESIGN AND PATIENTS: The study included 75 consecutive patients who had a shoulder MR scan followed by arthroscopy. MR images included oblique coronal, oblique sagittal (4 mm thick, 1 mm interslice gap), and angled oblique sagittal (3 mm/0.2 mm) images perpendicular to the lateral cuff. A musculoskeletal staff radiologist and fellow separately evaluated the cuff for tears on the oblique coronal images supplemented with either the oblique sagittal or the angled sagittal images. RESULTS: For distinguishing a cuff tear from no tear, the staff radiologist had an accuracy of 0.76 (95% confidence interval: 0.67, 0.85) with the standard sagittal set, and 0.88 (0.80, 0.95) with the angled set (P=0.04). There was a nonsignificant improvement in accuracy for the fellow, calculated as 0.73 (0.63, 0.83) on the standard sagittal set and 0.76 (0.67, 0.85) on the angled set. Both readers also improved their diagnostic accuracy for partial-thickness tears with the angled set, although the improvement was statistically significant only for the staff radiologist. CONCLUSION: There is a slight improvement in accuracy for diagnosing rotator cuff tears, particularly partial-thickness tears, for the more experienced radiologist using thinner-section angled oblique sagittal images. These images may be useful as a supplemental sequence in patients where it is important to identify partial thickness tears accurately. PMID- 11407718 TI - MR findings in athletes with pubalgia. AB - OBJECTIVE: To describe the MR findings in athletes with pubalgia. DESIGN AND PATIENTS: Pelvic MR images of 32 athletes (30 men, 2 women) with pubalgia were studied. T1-weighted and T2-weighted (SE and FSE) and STIR images in the axial and coronal planes were obtained on a 1.5-T system. Images were reviewed for general pelvic pathology. Special attention was given to the pubic symphysis, groin and pelvic musculature, and to the abdominal wall musculature. RESULTS: Thirty surgically confirmed cases comprise the study group. Abnormalities in the following were found: pubic symphysis (21/30), abdominal wall (27/30), groin musculature, including rectus abdominis (21/30), pectineus (6/30), and adductor muscle group (18/30). CONCLUSIONS: Pubalgia is a complex process which is frequently multifactorial. The MRI findings can alter the surgical approach. PMID- 11407719 TI - Pelvic instability after bone graft harvesting from posterior iliac crest: report of nine patients. AB - OBJECTIVE: To report the imaging findings in nine patients who developed pelvic instability after bone graft harvest from the posterior aspect of the iliac crest. DESIGN AND PATIENTS: A retrospective study was performed of the imaging studies of nine patients who developed pelvic pain after autologous bone graft was harvested from the posterior aspect of the ilium for spinal arthrodesis. Plain films, bone scans, and CT and MR examinations of the pelvis were reviewed. Pertinent aspects of the clinical history of these patients were noted, including age, gender and clinical symptoms. RESULTS: The age of the patients ranged from 52 to 77 years (average 69 years) and all were women. The bone graft had been derived from the posterior aspect of the iliac crest about the sacroiliac joint. All patients subsequently developed subluxation of the pubic symphysis. Eight patients had additional insufficiency fractures of the iliac crest adjacent to the bone graft donor site, and five patients also revealed subluxation of the sacroiliac joint. Two had insufficiency fractures of the sacrum and one had an additional fracture of the pubic ramus. CONCLUSIONS: Pelvic instability is a potential complication of bone graft harvesting from the posterior aspect of the iliac crest. The pelvic instability is manifested by insufficiency fractures of the ilium and subluxation of the sacroiliac joints and pubic symphysis. PMID- 11407720 TI - Bizarre parosteal osteochondromatous proliferation with cortical invasion. AB - A 15-year-old male presented with an 18-month history of increasing swelling on the radial aspect of his left forearm. Radiographs and MR images showed a partly calcified mass on the lateral border of the radius with erosion of the underlying cortex. Following a CT-guided needle biopsy, the mass was excised by segmental resection of the radial shaft with replacement by a vascularised fibular graft. Histological examination of the resected specimen showed many features typical of a bizarre parosteal osteochondromatous proliferation (BPOP) but with destruction of the underlying cortical bone by cellular spindle cell tissue. Although there were no cytological signs of malignancy, we believe that this unusual disease should be regarded as a particularly aggressive form of BPOP and treated by wide excision. Clinical follow-up of our patient at 2 years shows no sign of local recurrence or metastasis. PMID- 11407721 TI - Paravertebral neurinoma associated with aggressive intravertebral extension. AB - Neurinomas are relatively common benign tumors thought to arise from nerve sheath cells. Although intraosseous neurinomas may destroy the bone, extraosseous neurinomas with extensive destruction and invasion of bone are considered rare. We present two unusual cases of a benign extraosseous neurinoma that extensively invaded the vertebral body through the nutrient canal. PMID- 11407722 TI - Spinal epidural hemangioma related to pregnancy. AB - We report the case of a 39-year-old woman with adolescent idiopathic scoliosis presenting with myelopathy secondary to a spinal epidural hemangioma. MRI showed an epidural soft tissue mass within the spinal canal between T5 and T9 with severe spinal cord compression. Symptoms had a temporal relationship to her pregnancy. Surgical removal of the epidural hemangioma rapidly relieved her symptoms and neurologic deficits. Follow-up examination 2 years later demonstrated normal motor and sensory function, without any neurologic sequelae or progression of deformity. PMID- 11407723 TI - Calcification of the alar ligament of the cervical spine: imaging findings and clinical course. AB - Ligamentous calcification of the cervical spine has been reported in the yellow ligament, anterior and posterior longitudinal ligaments and interspinous ligament. Calcification in the upper cervical spine is rare, although some cases with calcification of the transverse ligament of the atlas have been reported. Two patients with calcification of the alar ligament with an unusual clinical presentation and course are described. Examination by tomography and computed tomography (CT) showed calcification of the alar ligament and the transverse ligament of the atlas. CT documented decreased calcification as symptoms resolved. There may be a role for CT in the search for calcifications in the upper cervical spine in patients presenting with neck pain and pharyngodynia if radiographs are normal. PMID- 11407724 TI - Evolution and stabilization of vulnerable atherosclerotic plaques. PMID- 11407725 TI - Clinical characteristics of patients with constrictive pericarditis after coronary bypass surgery. AB - Constrictive pericarditis (CP) is an unusual sequela of cardiac surgery, so the present study evaluated the clinical characteristics of patients with CP after coronary artery bypass grafting (CABG). Four hundred and sixty-three patients who underwent isolated CABG between January 1989 and March 1999 were examined retrospectively. The first choice of treatment for postoperative pericardial effusion was non-steroid anti-inflammatory agents, and an increased dose of diuretics. The second treatment choice was corticosteroids or pericardial drainage. When CP was suspected during the follow-up period (mean, 54+/-31 months), cardiac catheterization was carried out to establish the diagnosis. Of the 463 patients undergoing CABG, there were 11 (2.4%) who developed CP after surgery. The median time to the onset of symptoms after CABG was 4 weeks (range, 3-96 weeks). On univariate and multivariate analysis, normal left ventricular ejection fraction, warfarin administration, and early postoperative pericardial effusion were significantly associated with a greater potential of postoperative CP. The effusion was bloody in all cases of pericardial drainage despite warfarin therapy. Not draining the postoperative effusive pericardial effusion was a risk factor for the development of CP. Pericardial drainage for patients with significant effusion after CABG is important for the prevention of subsequent CP, especially in those patients being treated with warfarin or with normal left ventricular function. PMID- 11407726 TI - Arrhythmias in patients with Brugada-type electrocardiographic findings. AB - Brugada syndrome is characterized by marked ST-segment elevation in the right precordial leads (Bru-ECG) and is associated with a high risk for sudden death. However, it is unclear whether the arrhythmogenesis is caused by the mechanisms responsible for Bru-ECG. The present study investigated the risk of arrhythmias in patients with Bru-ECG by retrospectively analyzing 30 patients (28 men; mean age, 51+/-14 years) with Bru-ECG. Aborted sudden cardiac death (ventricular fibrillation or syncope) occurred in 9 patients (30%); paroxysmal atrial fibrillation was present in 9 (30%) patients in addition to malignant ventricular arrhythmias, and some type of arrhythmic event (aborted sudden cardiac death or paroxysmal atrial fibrillation) occurred in 15 patients (50%). Of all the arrhythmic events, 93% occurred at night or early in the morning, and 92% had pronounced ST-segment elevation. These results suggest that Bru-ECG may be associated not only with an increased risk of ventricular tachyarrhythmias but also with an increased risk of paroxysmal atrial fibrillation, and that the arrhythmogenesis may be related to the pronounced ST-segment elevation. PMID- 11407727 TI - Beneficial effect of troglitazone, an insulin-sensitizing antidiabetic agent, on coronary circulation in patients with non-insulin-dependent diabetes mellitus. AB - Evidence is increasing for small vessel remodeling and disturbance of endothelium dependent vasodilation in diabetic patients. Insulin increases vascular wall thickening and produces endothelial dysfunction. Troglitazone, a new insulin sensitizer antidiabetic agent, is considered to reduce plasma insulin level and the present study assessed its effect on the coronary circulation of the patients with non-insulin-dependent diabetes mellitus (NIDDM). Analysis of the myocardial washout rate with adenosine triphosphate-stress thallium-201 scintigraphy was used to estimate coronary circulation, and for estimation of insulin sensitivity, the homeostasis model insulin resistance index (HOMA-R) was calculated. Patients were treated with monotherapy of either troglitazone (200 mg bid, n=12) or glibenclamide (2.5 mg daily, n=12) for 3 months. Age-, sex- and risk factors matched subjects without NIDDM were employed as a control. Fasting plasma glucose and hemoglobin A1c were similarly decreased by troglitazone or glibenclamide. Plasma insulin level (pmol/L) decreased from 66.6+/-10.8 to 39.0+/-7.2 with troglitazone, but was unchanged by glibenclamide (58.8+/-7.2 to 66.0+/-10.8). The diabetic groups had a significantly lower washout rate than controls, which was improved by troglitazone, but not by glibenclamide. In addition, the increase in washout rate correlated significantly with the decrease in HOMA-R in the troglitazone group. In conclusion, troglitazone can restore coronary circulation by improving insulin resistance in patients with NIDDM. PMID- 11407728 TI - Serum thioredoxin (TRX) levels in patients with heart failure. AB - An increase in oxidative stress is thought to be involved in the progression of heart disease, but the serum level of thioredoxin (TRX), which regulates the cellular redox state, has not been investigated in patients with heart diseases. The present study determined serum TRX levels with a sandwich enzyme-linked immunosorbent assay in a total of 39 patients with dilated cardiomyopathy (DCM) (n=5), acute coronary syndrome (ACS) (n=7) or stable angina (n=18), including effort angina (n=7) and vasospastic angina (n=11), and in control subjects (n=7). The serum TRX level in patients with New York Heart Association (NYHA) functional classes III and IV (n=8, 33.3+/-8.6 ng/ml) was significantly higher than in the control subjects (n=7, 14.0+/-4.6 ng/ml). In addition, the serum TRX levels correlated positively with the severity of NYHA class, and negatively with the left ventricular ejection fraction. The serum TRX levels were elevated in patients with ACS and DCM compared with the controls. These results indicate a possible association between TRX concentration and the severity of heart failure. PMID- 11407729 TI - Lipid peroxidation may predict restenosis after coronary balloon angioplasty. AB - The present study assessed whether lipid peroxidation in plasma might predict restenosis after coronary balloon angioplasty. A total of 87 patients, who had undergone successful coronary balloon angioplasty using standard techniques, were enrolled. Fasting blood samples before the intervention were measured for plasma levels of thiobarbituric acid reactive substances (TBARS, an indicator of lipid peroxidation). Angiography was carried out before and 15 min after angioplasty, and at follow-up (4 months after angioplasty), and evaluated using a quantitative approach. There were 23 patients with restenosis (group R) and 64 patients without restenosis (group N) after coronary balloon angioplasty. The plasma TBARS level (mean+/-SEM) of 4.3+/-0.1 micromol/L in group R was significantly higher than that of 3.2+/-0.1 micromol/L in group N (p<0.01). There were no significant differences in other parameters, including plasma lipid levels, between the 2 groups. The plasma level of TBARS positively correlated with lumen loss of the coronary artery at the time of follow-up angiography (r=0.57, p<0.01). Our results suggest that oxidative stress contributes to restenosis and indicate that an elevated plasma level of TBARS may be a reliable predictor of restenosis. PMID- 11407730 TI - Imbalance of cusp width and aortic regurgitation associated with aortic cusp prolapse in ventricular septal defect. AB - The Doppler echocardiograms of the aortic valve and associated aortic regurgitation (AR) were reviewed in 72 patients with a ventricular septal defect (VSD). Group I comprised 13 patients without any deformity of the aortic cusp for > or = 10 years, group 2 included 35 patients who did not develop AR for > or = 10 years after right coronary cusp prolapse (RCCP) was first detected, group 3 comprised 11 patients with RCCP and AR in whom the AR remained subclinical for > or = 10 years, and group 4 was 13 patients who underwent surgical treatment because of moderate to severe AR. The cusp imbalance index [width of right (R) or non- (N) coronary cusp/width of left coronary cusp (L)] was compared among the 4 groups. R/L or N/L was larger in group 4 than in groups 1-3; R/L exceeded 1.30 in all the patients in group 4, whereas it was less than 1.30 in all the atients in groups 1-3. Two patients in group 4 with non-coronary cusp prolapse had an N/L greater than 1.50. No other patients in any group had an N/L larger than 1.20. An imbalance of cusp width may predict possible progressive deterioration of AR. PMID- 11407731 TI - Histopathologic evaluation of coronary artery thrombi obtained by directional coronary atherectomy in patients with restenosis-induced unstable angina pectoris. AB - The pathogenesis of unstable angina pectoris (UAP) following percutaneous transluminal coronary angioplasty (PTCA) or directional coronary atherectomy (DCA) has not been adequately investigated, so the present study aimed to determine whether thrombi are present in restenotic lesions. The study group comprised 14 patients (16 arterial branches) with angina pectoris in whom either PTCA or DCA was performed and who had developed UAP associated with restenosis, and who then underwent DCA of the restenosed lesion (R-UAP group). The control groups comprised individuals with UAP undergoing DCA with no prior history of PTCA or DCA (P-UAP group; n=29, 29 branches), patients with acute myocardial infarction (AMI group; n=34, 34 branches), and patients with stable angina pectoris (SAP group; n=31, 33 branches). The presence of thrombi was determined by light microscopy of histologic specimens. Thrombus was present in only 1 of the 16 (6.3%) branches in the R-UAP group. 21 of the 29 (72.4%) branches in the P UAP group, and in 25 of the 34 (73.5%) in the AMI group. In the SAP group, it was detected in only 2 of the 33 (7.1%) branches. The incidence of thrombus was significantly lower in the R-UAP group than in the P-UAP group. In conclusion, the role of thrombus is limited in causing post-interventional UAP at restenosed sites. PMID- 11407732 TI - Association between job strain status and cardiovascular risk in a population of Taiwanese white-collar workers. AB - Using data from a survey of a white-collar working population in Taiwan (438 women, 526 men), the relation between job strain status and cardiovascular risk factors (high serum total cholesterol, low serum high-density lipoprotein (HDL) cholesterol, and high plasma fibrinogen) was examined. Job strain indicators, defined by Karasek's model, included psychological demand and decision latitude. Blood pressure, cholesterol and fibrinogen were analyzed as continuous variables, whereas psychological demand and decision latitude were dichotomized into 2 levels and job strain into 4 exposure categories. Plasma fibrinogen was significantly and positively associated with job strain status in both male and female workers and also with decision latitude in female workers only. No consistent association between job strain status and total serum and HDL cholesterol was detectable. In conclusion, plasma fibrinogen is a possible intermediate factor linking occupational stress to elevated cardiovascular risk. PMID- 11407733 TI - Using isoproterenol stress echocardiography to predict the response to carvedilol in patients with dilated cardiomyopathy. AB - Trials have demonstrated that carvedilol can produce hemodynamic, symptomatic, and prognostic improvements in dilated cardiomyopathy (DCM), but some DCM patients have deteriorated after carvedilol, developing congestive heart failure. The present study investigated the use of isoproterenol (ISP) stress echocardiography to select those patients with DCM who would respond to carvedilol. ISP was infused intravenously in 22 patients with DCM and they were classified into 2 groups based on the left ventricular systolic response: good response to ISP [change in fractional shortening (FS) with ISP > 0.05, n=13] and poor response to ISP (change < or = 0.05, n=9). In the good response group, FS significantly increased from 0.12+/-0.04 to 0.17+/-0.08 (mean+/-SD, p<0.05) with carvedilol, and 7 patients improved symptomatically (New York Heart Association class). However, in the poor response group, no significant difference was observed between FS at baseline and that at the end of follow-up. Moreover, only 1 patient in the poor response group improved symptomatically. ISP stress echocardiography can assist in selecting patients with DCM who will respond positively to carvedilol. PMID- 11407734 TI - Polymorphic ventricular tachycardia in patients with vasospastic angina--clinical and electrocardiographic characteristics and long-term outcome. AB - There have been few clinical studies exploring the characteristics of spontaneous polymorphic ventricular tachycardia (VT) during a vasospastic angina attack. During a 4-year recruitment period, Holter ECG recordings were monitored for 42+/ 24 h during a drug-free period in 60 consecutive patients with vasospastic angina (VSA) and of these, 8 patients had at least one episode of polymorphic VT during monitoring. Ischemic ST segment elevation was immediately preceded the spontaneous polymorphic VT in all 8 patients, 4 of whom had silent coronary vasospasm. Immediately before the onset of polymorphic VT, both R-on-T and long short sequences were observed in 4 of the 8 patients and ST wave alternans were recorded in 2 patients. VT exhibited a pattern of torsade de pointes in 4 of the 8 patients. Five patients underwent electrophysiologic testing during a drug-free asymptomatic phase, and polymorphic VT was induced in 2 of the 5 patients, with one developing ventricular fibrillation. During a follow-up period of 73+/-17 months, there was a significant difference in the incidence of sudden death between patients with and without VT (2/8 cases [25%] vs 0/52 [0%]; p<0.01). Thus, vasospastic attacks, even if asymptomatic, that immediately precede the development of polymorphic VT may be associated with a repolarization abnormality and an increased risk of sudden death. PMID- 11407735 TI - Nicorandil, a hybrid between nitrate and ATP-sensitive potassium channel opener, preconditions human heart to ischemia during percutaneous transluminal coronary angioplasty. AB - The human heart progressively becomes more tolerant to ischemia after repeated balloon inflations during percutaneous transluminal coronary angioplasty (PTCA). The present study investigated whether nicorandil, a hybrid between nitrate and an ATP-sensitive potassium channel opener, affects this ischemic preconditioning. Sixteen patients with stable angina pectoris caused by left anterior descending artery lesions were subjected to 2 balloon inflations of 2-min duration with a 3 min reperfusion period. Seven of these patients served as the control group and in the remaining 9 patients, nicorandil was administered intravenously (6 mg/h) throughout the PTCA procedure (nicorandil group). The lactate extraction ratio (LER) was obtained at 30 s after each ischemic event (LERpost-1 and LERpost-2) in both groups. In the control group, LERpost-1 was more negative than LERpost-2 ( 185.7+/-74.2 vs -98.0+/-37.3%, p<0.01). The ratio of the sum of the ST elevation in the precordial leads during the second inflation (sumST-2, 0.94+/-0.66 mV) to that during the first inflation (sumST-1, 1.43+/-1.17 mV) was 0.72+/-0.16 in the control group, which was less than the ratio in the nicorandil group (1.06+/ 0.13, p<0.01). Nicorandil abolished the difference between the 2 ischemic events (LERpost-1, -45.1+/-41.6 vs LERpost-2, -43.5+/-51.1%; sumST-1, 1.38+/-0.80 vs sumST-2, 1.46+/-0.90 mV). LER was less negative in the nicorandil group than that in the control group (LERpost-1, -45.1+/-41.6 vs -185.7+/-74.2%, p<0.01; LERpost 2, -43.5+/-51.1 vs -98.0+/-37.3%, p<0.05). Thus, nicorandil improved lactate metabolism during PTCA without significantly influencing ST-elevation. In conclusion, intravenous pre-administration of nicorandil appears to precondition the human heart during PTCA. PMID- 11407736 TI - Antiarrhythmic drug, cibenzoline, can directly improve the left ventricular diastolic function in patients with hypertrophic cardiomyopathy. AB - The effect of cibenzoline on left ventricular diastolic function was investigated in patients with hypertrophic cardiomyopathy (HCM). Before and 2 h after an oral administration of 200 mg of cibenzoline, echocardiographic, apexcardiographic and gated radionuclide angiographic studies were performed in 12 patients with hypertrophic obstructive cardiomyopathy (HOCM) and 7 with hypertrophic nonobstructive cardiomyopathy (HNCM). After administration of cibenzoline, the left ventricular pressure gradient decreased from 96+/-33 mmHg to 29+/-22 mmHg (<0.0001). Fractional shortening decreased from 53.3+/-7.5 to 45.4+/-6.2% (p=0.0008) in patients with HOCM and from 49.9+/-8.7 to 40.9+/-7.5% (p=0.0039) in patients with HNCM. On the other hand, E-wave velocity increased and A-wave velocity decreased in both groups. The time between the second heart sound and O point was shortened from 253+/-53 to 176+/-21 ms (p<0.0001) in patients with HOCM and from 245+/-54 to 185+/-44 ms (p=0.0050) in patients with HNCM. The time to peak filling rate was shortened from 248+/-79 to 190+/-40 ms (p=0.0072) in patients with HOCM and from 218+/-33 to 163+/-26 ms (p=0.0052) in patients with HNCM. These results indicate that in patients with HCM, cibenzoline suppresses left ventricular systolic function, but can markedly improve left ventricular diastolic dysfunction through its direct action. PMID- 11407737 TI - Clinical significance of preheparin serum lipoprotein lipase mass in coronary vasospasm. AB - The present study investigated the clinical significance of preheparin serum lipoprotein lipase (LPL) mass in coronary vasospasm by examining its relationship with the acetylcholine-induced coronary artery response in patients without angiographically demonstrable atherosclerotic coronary artery disease (CAD). The subjects were 39 men who had suspected CAD and who underwent coronary angiography. Coronary vasospasm was defined as a marked luminal narrowing or total occlusion provoked by the intracoronary administration of acetylcholine. Preheparin LPL mass was lower (p<0.05) in 25 subjects in whom vasospasm was induced by the acetylcholine provocation test than in the 14 subjects with a negative response. As regards preheparin LPL mass, the subjects with multiple vessel spasm had significantly low concentrations (p<0.05) compared with single vessel spasm, although serum lipid levels were not significantly different. Multiple regression analysis revealed only preheparin LPL mass had a significant absolute t-value (2.016) among the coronary risk factors. Low preheparin LPL mass is interpreted as reflecting an impaired acetylcholine-induced coronary relaxation in coronary vasospasm and preheparin LPL mass may be useful as a marker of early stage coronary atherosclerosis that is not detectable by angiography. PMID- 11407738 TI - Monophasic transmitral flow pattern with less increase in heart rate indicates left ventricular dysfunction. AB - When heart rate (HR) increases, mitral flow can become monophasic. Prolonged isovolumic contraction and relaxation time (ICT and IRT), directly related to left ventricular (LV) function, can potentially influence the HR with monophasic mitral flow. The present study investigated the relation between HR that causes monophasic flow and LV function. During diagnostic catheterization, HR was increased using right atrial pacing by 2 beats/min every 2 min in a stepwise manner until the development of monophasic mitral flow in 17 patients with normal sinus rhythm. ICT, IRT, end-diastolic and end-systolic LV volumes, LV ejection fraction, LV peak + and -dP/dt, peak (+dP/dt)/P, and the relaxation time constant (tau) were measured by Doppler echocardiography or catheterization when monophasic mitral flow developed. The monophasic HR varied from 74 to 106 beats/min. By univariate analysis, ICT (p<0.01, r2=0.73), LV peak +dP/dt (p<0.05, r2=0.37), peak (+dP/dt)/P (p<0.01, r2=0.71), peak -dP/dt (p<0.05, r2=0.25), and tau (p<0.05, r2=0.33) had a significant correlation with monophasic HR. By multivariate analysis, prolonged ICT and reduced LV peak -dP/dt independently contributed to monophasic mitral flow with less increase in HR. Monophasic mitral flow with less increase in HR indicates impaired LV systolic and diastolic function during isovolumic contraction and relaxation. PMID- 11407739 TI - Effects of exercise training on myocardial fatty acid metabolism in rats with depressed cardiac function induced by transient ischemia. AB - The effects of exercise training on metabolic and functional recovery after myocardial transient ischemia were investigated in a rat model. Male Wistar Kyoto rats were subjected either to a 30-min left coronary artery occlusion followed by reperfusion or to a sham operation. At 4 weeks after operation, the rats were randomly assigned either to sedentary conditions or to exercise training for 6 weeks. In the ischemic rats, pinhole SPECT (single photon emission computed tomography) imaging with thallium-201 (201Tl) and 123I-(rho-iodophenyl)-3-R,S methylpentadecanoic acid (BMIPP) showed a reduction of both myocardial perfusion and fatty acid metabolism in the risk zone of the left ventricle (LV). The LV was dilated and the ejection fraction was decreased after ischemic injury. The severity score showed a significant decrease on both 201Tl and BMIPP (201Tl, from 19.9+/-2.7 to 17.0+/-2.2, p<0.05; BMIPP, from 21.5+/-2.4 to 18.6+/-1.9, p<0.05) after exercise training in the ischemic trained rats, but did not change significantly in their sedentary counterparts. Plasma levels of free fatty acids normalized in the ischemic trained rats, but elevated in the ischemic sedentary rats (0.53+/-0.05 vs 0.73+/-0.06 mmol/L, p<0.05). Furthermore, the trained rats had a significant increase in LV stroke volume (0.25+/-0.02 vs 0.21+/-0.01 ml/beat, p<0.05) and adaptive cardiac hypertrophy. These findings demonstrate that adaptive improvements in myocardial perfusion, fatty-acid metabolism and LV function were induced by exercise training after transient ischemia. PMID- 11407740 TI - Apoptosis of vascular smooth muscle cells is induced by Fas ligand derived from endothelial cells. AB - Although Fas-mediated cell death may play a role in atherogenesis, causal data in support of this hypothesis are lacking. The present study investigated the possibility that endothelial cells are involved in vascular smooth muscle cell (VSMC) apoptosis via the Fas-FasL pathway, and hence in atherogenesis. FACS analysis detected FasL on the surface of human umbilical vein endothelial cells (HUVECs) and immunofluorescence staining of the HUVECs demonstrated high levels of FasL in the intracellular compartment. FasL was down-regulated 4 h after tumor necrosis factor (TNFalpha) treatment, coinciding with maximal surface expression of the adhesion molecules vascular cell adhesion molecule-1 and E-selectin. However, the down-regulation of FasL expression was transient, as surface expression returned within 24 h of TNFalpha treatment. When cocultured with VSMCs, the FasL-expressing EC could kill the VSMCs in a manner that could be blocked by recombinant Fas-Fc, deployed as a soluble receptor for Fas. Moreover, when human coronary arteries were studied with immunohistochemistry using G247-4 monoclonal antibody for the detection of FasL, few FasL positive EC were observed in diffuse intimal thickening. In contrast, endothelium overlying the plaque showed prominent and uniform expression of FasL. These findings suggest that the Fas/FasL pathway can be used by EC to induce VSMC apoptosis in the atherosclerotic lesion. PMID- 11407741 TI - Difference in thioredoxin expression in viral myocarditis in inbred strains of mice. AB - Redox regulating mechanisms may be involved in the pathogenesis of viral myocarditis and thioredoxin (TRX) is a small multifunctional protein that contains a redox active sequence. The present study investigated the histopathology and characteristics of TRX expression in acute coxsackievirus B3 myocarditis in inbred strains of mice (severe myocarditis in DBA/2 mice, moderate myocarditis in BALB/c mice and mild myocarditis in C57BL/6 mice). Thioredoxin was upregulated and its expression correlated with the severity of the disease. In addition, 8-hydroxy-2'-deoxyguanosine, which is an established marker for oxidative stress, was concominantly positive in damaged myocytes. Thus, TRX may be specifically induced by the acute inflammatory stimuli in murine viral myocarditis, and the severity and development of acute viral myocarditis may be regulated by the cellular redox state. PMID- 11407742 TI - Potential use of ultrasound in creating transmyocardial channels. AB - Although the mechanisms of the clinical benefits of transmyocardial laser revascularization (TMLR) are considered to be angiogenesis with increased perfusion, denervation and placebo effect, it is unknown whether laser energy is a prerequisite in obtaining these beneficial effects. The present study investigated whether it is possible to create transmyocardial channels and induce angiogenesis by ultrasound. Myocardium was penetrated with an ultrasonically activated surgical blade by advancing the blade tip perpendicularly to the left ventricular free wall of the beating heart of 6 mongrel dogs. The power of ultrasound was set at either the lowest or highest of the system. The animals were killed 30 min (acute; n=3) and 2 weeks (chronic; n=3) after channel creation. Holmium:YAG laser, which is currently used for clinical TMLR, was used to create myocardial channels in 4 other dogs, which were also killed 30 min (n=2) and 2 weeks (n=2) after channel creation. The areas of acute channel core, acute thermal damage and chronic fibrosis were compared between the laser and ultrasound channels by Masson's trichrome stain. Factor VIII and proliferating cell nuclear antigen (PCNA) immunostaining were carried out on the samples obtained from chronic animals. The density of vessels and that of proliferating vascular endothelial cells and vascular smooth muscle cells around the channels were measured. The area of acute core was larger in the lowest and highest outputs of ultrasound than in laser channels (0.78+/-0.09, 1.0+/-0.12 vs 0.38+/ 0.04 mm2; p<0.01). The area of acute damage in both laser and the highest output of ultrasound channels was greater than in the channels produced by the lowest output of ultrasound (4.43+/-0.28, 4.63+/-0.44 vs 2.90+/-0.29 mm2; p<0.01). The ratio of acute damage area to acute core area was greater in laser channels than in either type of ultrasound channel (16.86+/-1.66 vs 6.04+/-0.67, 7.86+/-1.07; p<0.01) and the area of chronic fibrosis was greater (3.23+/-0.20 vs 1.59+/-0.18, 2.24+/-0.20 mm2; p<0.01). Factor VIII and PCNA immunostaining revealed new vessels not only inside the areas of chronic fibrosis, but also in the surrounding myocardium, in both laser and ultrasound channels. Ultrasound created transmyocardial channels histologically similar to laser channels and angiogenesis was induced in the normal myocardium surrounding ultrasound channels. PMID- 11407743 TI - Large thrombus in the ascending aorta successfully treated by thrombolysis--an unusual cause of acute massive myocardial infarction. AB - A 52-year-old woman suffered from acute massive myocardial infarction in association with a large thrombus in the ascending aorta. She was a moderate smoker and was taking hormone supplement therapy for menopausal hormone insufficiency and the contraceptive pill for endometriosis. Cardiac angiography revealed a large mobile filling defect close to the orifice of the left coronary artery, but the left coronary artery could not be visualized. Her hemodynamic condition was impaired so greatly that intraarterial counterpulsation and intravenous thrombolysis was immediately performed. The thrombus dissolved in 1 h and recanalization of the left coronary artery was achieved without serious systemic thromboembolism. She has been doing well with no cardiac events for 7 years. This is the second report of a large thrombus in ascending aorta being the cause of acute myocardial infarction in the whole territory of the left coronary artery, and the first to diagnose such a thrombus antemortem and treat it successfully. PMID- 11407744 TI - Utility of three-dimensional volume rendering images using electron-beam computed tomography to evaluate possible causes of ischemia from an anomalous origin of the right coronary artery from the left sinus of valsalva. AB - The present study evaluated the usefulness of 3-dimensional volume rendering (VR) images using electron-beam computed tomography (EBCT) in determining the possible causes of ischemia resulting from the anomalous origin of the right coronary artery (RCA) from the left sinus of Valsalva, which coursed between the ascending aorta and pulmonary trunk. Such anomalies could cause ischemia or sudden death without obstructive coronary artery disease. The suggested mechanism is either compression causing closure of the slit-like orifice of the anomalous artery as the aorta dilates with exertion or compression of the anomalous artery by the aorta and pulmonary trunk as it courses between these 2 arteries, which dilate with exercise. A 17-year-old male underwent EBCT coupled with a 100-ml intravenous injection of iodinated contrast medium. Data were reconstructed into 3-dimensional images through VR to evaluate the shape of the orifice and the spatial relationship of the RCA, ascending aorta and pulmonary trunk. Perspective VR showed the shape of the orifice of the left main trunk, which was not slit like, and cut-plane VR showed the spatial relationship of both the lumen and the surface of the RCA, ascending aorta and pulmonary trunk, providing information on whether the ascending aorta or pulmonary trunk would compress the RCA and cause ischemia. PMID- 11407745 TI - Successful removal of a left main coronary artery thrombus induced by vasospasm to the aorta [correction of vasospasm of the aorta] after the injection of contrast medium. AB - A 51-year-old woman with acute myocardial infarction underwent emergency coronary angiography. The patient had an episode of idiopathic thrombocytopenic purpura, and underwent splenectomy at the age of 36. On admission, platelet count in the peripheral blood was 43.2x10(4)/microl. During the initial emergency left coronary angiography, a thrombus was detected in the left main trunk. The thrombus was drawn back to the orifice of the left coronary artery, and finally disappeared into the aorta by injecting contrast media. After the thrombus disappeared, no stenotic lesion was detected in the left coronary artery. One month later, when acetylcholine-provocation coronary angiography was performed, marked vasospasm was detected in the left coronary arteries. Coronary thrombosis in the patient might have been induced by a coronary spasm, and the presence of thrombocytosis might also have affected the development of a coronary thrombus. However, it was spontaneously drawn back to the aorta by back flow of contrast media, which was injected via the entrance of the left coronary artery. A case of acute myocardial infarction whose thrombus occluded the left main coronary artery and was removed at first injection of contrast media is presented. PMID- 11407746 TI - Repair of left ventricular rupture following mitral valve replacement concomitant with left atrial reduction procedure--intracardiac patch and extracardiac buttress suture. AB - Rupture of the posterior wall of the left ventricle after mitral valve replacement is a dire complication associated with a very high mortality. This study reports a successful repair of type I left ventricular rupture, which occurred after mitral valve replacement concomitant with a left atrial reduction procedure, by combination of an intracardiac patch and an extracardiac buttress suture. In a case such as this, in which hemostasis is quite difficult to establish, this combination technique is particularly effective. PMID- 11407747 TI - Proinflammatory cytokine inhibitor prolongs the survival of rats with heart failure induced by pressure overload. AB - Although an increased expression of proinflammatory cytokines has been reported in cardiac tissue samples from patients with congestive heart failure (CHF) and in various animal models of CHF, the role of these cytokines in the disease remains to be determined. Dahl salt-sensitive (DS) rats fed a high salt diet develop hypertension, cardiac hypertrophy and eventually CHF. In the present study, DS rats were treated with FR167653 (1-[7-(4-fluorophenyl)-1,2,3,4 tetrahydro-8-(4-pyridyl)pyrazolo[5,1-c][1,2,4]triazin-2-yl]-2-phenylethanedione sulfate monohydrate), a new low molecular weight inflammatory cytokine inhibitor. Treatment with 10 mg/kg per day of FR167653 significantly prolonged the survival of the animals and also prevented the bodyweight loss associated with heart failure. In conclusion, a non-peptide proinflammatory cytokine inhibitor improved the survival of animals with heart failure. PMID- 11407748 TI - Evaluation of e-beam, gamma- and X-ray treatment on the chemistry and safety of polymers used with pre-packaged irradiated foods: a review. AB - Polymers used to package food intended for irradiation must currently receive separate US FDA approvals for e-beam, gamma and X-radiation. The three forms of irradiation have virtually indistinguishable effects on polymers held in vacuum. However, in air, irradiation damage is favoured by slow dose rates, such as that characteristic of gamma irradiation. Irrespective of irradiation type, theory and existing polymer irradiation literature both suggest that radiolytic products in food contact polymers should not present a health hazard due to migration into the foods they contain. Equations are presented which calculate the FDA allowable content of radiolytic products to achieve 'threshold of regulation' clearance and conversion of radiolytic yield values (G-values) into ppb concentration in the treated polymer in conformance with FDA practice. An approach to quantifying the functional barrier of a polymer is discussed. PMID- 11407749 TI - ICP/MS and ICP/AES elemental analysis (38 elements) of edible wild mushrooms growing in Poland. AB - Thirty-eight elements, including toxic cadmium, lead, mercury, silver and thallium, were determined in 18 species of wild edible mushrooms collected from several sites in Pomorskie Voivodeship in northern Poland in 1994. Elements were determined by double focused high resolution inductively coupled plasma mass spectrometry (HR-ICP-MS) and inductively coupled plasma atomic emission spectroscopy (ICP-AES), after wet digestion of the dried samples with concentrated nitric acid in closed PTFE vessels using a microwave oven. K, P and Mg were present at levels of mg/g dry matter; Na, Zn, Ca, Fe, Cu, Mn, Rb, Ag, Cd, Hg, Pb, Cs, Sr, Al and Si were present at microg/g levels, while Tl, In, Bi, Th, U, Ce, Pr, Nd, Sm, Eu, Gd, Tb, Dy, Ho, Er, Tm, Yb, La, Lu and Ba were present at ng/g levels. PMID- 11407750 TI - The influence of complexing agents on the solubility and absorption of aluminium in rats exposed to aluminium in water. AB - The influence of citrate (0-31 mM), fluoride (0 or 2.6 mM) and silicate (0 or 2.6 mM) on the absorption of Al (0-18 mM) was studied in rats. We tested the hypothesis that the solubility and absorption of Al increases in the gastrointestinal (GI) tract in the presence of the complexing agents. Male rats were exposed for 6 or 7 weeks to soluble Al in acidic drinking water (pH 2.5-3.0) with or without the complexing agents. At the end of exposure Al was fractionated in the stomach content, in order to study if the solubility of Al was changed after ingestion. Al absorption was estimated by Al analysis of the right femur bone. Speciation calculations indicated that citrate and fluoride caused formation of soluble Al-citrate (97%) and -fluoride (> 60%) complexes in the water. Silicate did not affect the theoretical speciation. In all cases, a large fraction of soluble Al became insoluble in the stomach after ingestion. The concentration of soluble Al increased only in the presence of citrate or a mixture of fluoride and silicate, but citrate was the only complexing agent that influenced the absorption of Al in the rat. This indicates that the form of Al may be changed in the GI tract when soluble drinking-water Al is ingested, and that the solubility of Al in drinking water and GI tract may not be good predictors of the bioavailability of Al even when chelating agents are present. PMID- 11407751 TI - Identification of anatoxins in blue-green algae food supplements using liquid chromatography-tandem mass spectrometry. AB - Blue-green algae (cyanobacteria) in tablets and capsules, which are marketed as health food supplements, were investigated for the presence of neurotoxins related to anatoxin-a. These neurotoxins, which are nicotinic agonists, were investigated using isocratic micro-liquid chromatograph-tandem mass spectrometry (micro-LC-MS-MS). The investigated compounds were anatoxin-a and homoanatoxin-a, together with their degradation products, dihydroanatoxin-a, epoxyanatoxin-a, dihydrohomoanatoxin-a and epoxyhomoanatoxin-a which were synthesized from the parent toxins. The analytes were extracted with methanol followed by isocratic chromatography on a micro C18 reversed-phase column using acetonitrile-water, 50:50 (v/v), containing 20 mm acetic acid at 30 microl min(-1). The toxins were ionized in an ionspray (IS) interface operating in the positive ion mode, where the intact protonated molecules, [M + H]+, were generated at m/z 166, m/z 168, m/z 182, m/z 180, m/z 182 and m/z 196, for anatoxin-a, dihydroanatoxin-a, epoxyanatoxin-a, homoanatoxin-a, dihydrohomoanatoxin-a and epoxyhomoanatoxin-a, respectively. These served as precursor ions for collision-induced-dissociation (CID) and diagnostic product ions for these anatoxins were identified to carry out toxin confirmation by selected reaction monitoring (SRM) LC-MS-MS analysis. Dihydrohomoanatoxin-a and a novel isomer of epoxyanatoxin-a were identified in blue-green algae tablets. This finding suggests that a potential human health hazard could be associated with the consumption of these food supplements. PMID- 11407752 TI - Occurrence of heterocyclic aromatic amines in the Swiss diet: analytical method, exposure estimation and risk assessment. AB - A total of 86 meat samples, prepared in restaurants or homes, ready to eat (including poultry and fish) and 16 commercial samples such as bouillon (cubes) were analysed for heterocyclic aromatic amines (HAA). The analytical method consisted of an acidic extraction, clean-up on a cation exchange cartridge followed by an analogous HPLC step to recover the following HAA: IQ, MeIQ, MeIQx, 4,8-DiMeIQx, PhIP and 7,8-DiMeIQx. The HAA containing HPLC-fractions were collected, the HAA identified and quantified using two RP-HPLC-systems of different retention properties (UV-detection). The limit of quantitation was in the range of 0.2-0.4 ng/g and the relative repeatability 6-15%. The recovery of PhIP was lower than for the other HAA analysed (less than 80%) and a correction factor was applied. No significant differences of the HAA-concentration were found in samples from homes and restaurants, half of the total samples contained HAA at the following frequencies: PhIP and MeIQx 33% (each), 4,8-DiMeIQx 11% and MeIQ 4%; 7,8-DiMeIQx and IQ were not detected. The frequencies in commercial products were for MeIQx 31%, 7,8-DiMeIQx 19%, IQ 13% and PhIP 6%; MeIQ and 4,8 DiMeIQx were not found. Based on these data, the average exposure of Swiss adults to HAA was estimated to be 5 ng/kg body mass per day, commercial products contributing less than 10%. The theoretical excess cancer risk due to this intake was estimated on the base of the carcinogenic potency of the HAA in long-term animal experiments by linear extrapolation. The resulting risk in the order of 10(-4) at the maximum is discussed in terms of Swiss epidemiological data. PMID- 11407753 TI - Aflatoxins B1 in different grades of chillies (Capsicum annum L.) in India as determined by indirect competitive-ELISA. AB - Samples of the three grades of chilli pod (grades 1 to 3) were collected during surveys in 1998 and 1999 from the principal market yards and cold storage facilities of the major chilli-growing areas of Andhra Pradesh (AP), India. Chilli powders were collected from different supermarkets in Hyderabad, AP. They were analysed for aflatoxin B1 (AFB1) content by an indirect competitive ELISA. To avoid the influence of interfering substances present in chilli extracts, it was necessary to prepare the aflatoxin standards in methanol extracts of chillies free from aflatoxins. For nine representative samples there was good agreement between ELISA and HPLC estimations of AFB1 and the results suggested that the ELISA procedure adopted was dependable. Of the 182 chilli samples tested, 59% of the samples were contaminated with AFB1 and 18% contained the toxin at non permissible levels. The highest AFB1 concentration of 969 microg/kg was found in one sample representing grade 3. Overall the maximum percentage of chilli pods showing AFB1 levels higher than 30 microg/kg (non-permissible levels) was in grade 3. Chilli pods stored in refrigerated rooms showed the lowest proportion of samples containing aflatoxin. Nearly 9% of the chilli powders sold in supermarkets contained non-permissible aflatoxin levels. This report highlights the importance of using grade 1 chilli pods to minimize aflatoxin contamination. PMID- 11407754 TI - Determination of ochratoxin A in pig liver-derived pates by high-performance liquid chromatography. AB - A solid phase extraction procedure followed by HPLC analysis with fluorescence detection has been developed for ochratoxin A (OTA) in pates derived from pig liver. After a previous extraction of OTA with 60% acetonitrile, all the samples were purified through C8 columns. The percentage recovery was 85.7% and the lower limits calculated for accurate detection and quantitation were 0.56 ng/g and 0.84 ng/g respectively. The HPLC procedure showed a good linearity in the interval of equivalent OTA concentrations of 0.84 to 3 ng/g. Values <10% were obtained for precision in HPLC determinations performed (n = 3) and (n = 9). Stability of calibration standards and samples during the analytical procedure was also demonstrated. This method was successfully applied to 38 pig-derived pates and three samples were found to be positive with OTA levels above the detection limit. The highest concentration (1.77 ng/g) has been found in a home-made pate. PMID- 11407755 TI - Ochratoxin A in beverages from Morocco: a preliminary survey. AB - In a preliminary study, samples of Moroccan wines (n = 30), beers (n = 5) and fruit juices (n = 14) were assayed for ochratoxin A (OTA) by HPLC with fluorimetric detection, followed by confirmation by cleavage of the OTA molecule using carboxypeptidase with HPLC-fluorimetric determination of ochratoxin alpha (OT alpha). All the wine samples were contaminated, and the overall median OTA concentration was 0.65 microg/l (range 0.028-3.24 microg/l). One of the 14 samples of fruit juices was contaminated with a concentration of 1.16 microg/l, whereas none of the five beer samples was contaminated. This is the first report on the occurrence of OTA in various beverages from Morocco. PMID- 11407756 TI - Di(2-ethylhexyl) phthalate contamination of retail packed lunches caused by PVC gloves used in the preparation of foods. AB - Plasticizer contamination of foods sold in retail packed lunches and set lunches in restaurants was determined by GC/MS. The phthalate esters were as follows: diethyl, dipropyl, dibutyl, dipentyl, dihexyl, butylbenzyl, dicyclohexyl, di(2 ethylhexyl), dioctyl, diisooctyl (mixture of isomers) and diisononyl (mixture). Di(2-ethylhexyl) adipate was also determined. Sixteen packed lunches and ten set lunches were analysed, and in all samples the concentration of di(2-ethylhexyl) phthalate (DEHP) was the highest, at 0.80-11.8 mg/kg in packed lunches and 0.012 0.30 mg/kg in set lunches. The DEHP content of five packed lunches exceeded 1.85 mg, which is the EU tolerable daily intake (TDI) for a person of 50 kg body weight. Foodstuffs that were components of the packed lunches were taken from the factory at each step of preparation and phthalates were determined. For example, chicken contained 0.08 mg/kg DEHP when uncooked, 13.1 mg/kg after frying and 16.9 mg/kg after packing. Disposable PVC gloves used in the preparation of foods were apparently the source of high DEHP concentrations. The gloves used during cooking or packaging were sprayed with 68% (w/w) ethanol to sterilize them. PVC gloves from the factory contained 22 or 41% by weight of DEHP. To confirm the link with the contamination problem, samples of boiled rice, croquette and boiled dry radish were handled in the laboratory with PVC gloves containing 30% (w/w) DEHP. DEHP migration levels of 0.05 mg/kg in rice or 0.33 mg/kg in croquette, and 11.1 mg/kg in radish were found. The alcohol sprayed onto the gloves increased the migration of DEHP to 2.03 mg/kg in rice, 2.45 mg, kg in croquette, and 18.4 mg/kg in radish. PMID- 11407757 TI - Phosphite additives and their transformation products in polyethylene packaging for gamma-irradiation. AB - Infrared (IR) spectroscopy has been used to identify and quantify the phosphite antioxidant, tris(2,4-di-tert.butylphenyl) phosphite, and its corresponding phosphate in high density polyethylene (HDPE) food trays, both in the original, commercial trays and also as a function of gamma-irradiation and post irradiation, storage conditions. This direct method of inhibitor analysis complements time-consuming (some times non-quantitative) extraction methods, which for organo-phosphites are complicated by their facile conversion to phosphates by peroxidic impurities in the solvent. Because of the complete destruction of phosphite to give mainly phosphate at quite low gamma-irradiation doses (approximately 5 kGy) and phosphate formation during melt processing and radiation sterilization of these HDPE trays must take into account products from the irradiation of phosphate. Any residual phosphite is lost progressively in post-irradiation reactions, which are complex, producing a less than-quantitative yield of phosphate. PMID- 11407758 TI - Metabolic regulation of pH in plant cells: role of cytoplasmic pH in defense reaction and secondary metabolism. AB - A new biochemical pH-stat hypothesis that revised the classic hypothesis is presented to understand the metabolic regulation of intracellular pH in plant cells. Alternative pathway glycolysis, alternative pathway respiration and malate derived lactic and alcoholic fermentation (alternative pathway fermentation), all unique to plants, are integrated into a regulatory mechanism of pH in the cytoplasm. Its uniqueness to plant kingdom is discussed from the evolutionary viewpoint: it is suggested that when the ancestors of extant terrestrial plants expanded their habitat from oceans to freshwater, they abandoned a "sodium system" and adopted a "proton system" for nutrient uptake. Validity of the new hypothesis is examined with available data on a secondary active transport, anoxia and other experimental evidence. The hypothesis predicts that biotic and abiotic stress-induced cytoplasmic acidification triggers synthesis of phytoalexins and other secondary metabolites. Possible roles of cyanide-resistant alternative pathway respiration in the secondary metabolite production, metabolic switching between primary and secondary metabolisms, and defense reactions are proposed. PMID- 11407759 TI - Genetic regulation of gravitropism in higher plants. AB - Gravitropism is a classical subject in plant physiology. However, the molecular mechanisms that regulate gravitropism are unknown. Recently, many gravitropic mutants have been isolated from Arabidopsis thaliana and several genes for gravitropism have been cloned and characterized. These studies have shown that (1) the endodermis is essential for shoot gravitropism and (2) an auxin transport system and signaling pathway are necessary for gravitropism. Recent studies in Arabidopsis are reviewed and genetic regulation of gravitropism in this organism is discussed. PMID- 11407760 TI - Biology of kidney cells: ontogeny-recapitulating phylogeny. AB - Biology of kidney cells can be used as a model for further understanding of ontogeny-recapitulating phylogeny. The common and species-specific structural and functional relationship between blood capillaries and the environment via a filtration barrier of nephrons is a biological phenomenon resulting from renal cell memory acquired through evolution. Genetically programmed development, a subsequent series of gene expression, and inductive interactions played a key role in differentiation and maintenance of specific activities of kidneys in birds and mammals. Various environmental factors may alter kidney development and specific activities at the levels of gene expression, repression, or derepression, and defensive mechanisms involved in reaction to risk factors are developed. Autoimmunity and cancerogenesis are closely dependent on a variety of environmental agents, such as antigens originating from infections with some viruses and toxins, or irradiation, advanced industrialization, and progress of civilization. As a result of gene mutation, delation, rearrangement, and/or susceptibility to different agents, renal cell memory is altered. Instead of cell specific activities, the abilities for regeneration, and other genetically programmed activities, the genesis of kidney diseases are common. Balkan endemic nephropathy, as regional disease, is an important example of the role, of environmental agents, at the level of genes. Research programs on molecular genetics will contribute to our efforts both to prevent infections and to elucidate the genesis, diagnosis, prognosis, prevention, and therapy of kidney diseases. PMID- 11407761 TI - Circadian rhythms in microalgae. AB - Circadian rhythms have been described in a variety of microalgae. In each group, some model organisms arose and most detailed studies have been done with them. They include the cyanobacterium ("blue-green alga") Synechococcus and eukaryotic microalgae Gonyaulax polyedra (Dinophyta), Chlamydomonas reinhardtii (Chlorophyta), and Euglena gracilis (Euglenophyta). This review focuses on recent approaches to depict molecular components of the circadian system and the mechanisms of regulation in these organisms. In Synechococcus, the identification of the kailocus, which represents a central part of its oscillatory system, is discussed, as well as diverse approaches based on a luminescent reporter gene, which is driven by a clock-controlled cyanobacterial promoter. In eukaryotic microalgae, the diversity of genes/proteins that are controlled by the circadian clock is described and the kind of regulation (transcriptional and translational control) is emphasized. The role and function of conserved clock-controlled RNA binding proteins such as CCTR from Gonyaulaxor Chlamy 1 from Chlamydomonas are discussed. PMID- 11407762 TI - Proadrenomedullin-derived peptides in the paracrine control of the hypothalamo pituitary-adrenal axis. AB - Adrenomedullin (ADM) and proadrenomedullin N-terminal 20 peptide (PAMP) are widely distributed in various body tissues and organs, including the hypothalamo pituitary-adrenal (HPA) axis. ADM and PAMP inhibit in vitro release of ACTH from pituitary corticotropes, and findings suggest that this effect may become relevant when an exceedingly high ACTH secretion must be counteracted. ADM directly supresses angiotensin-II- and K+-stimulated aldosterone secretion from ZG cells, acting through calcitonin gene-related peptide (CGRP) type 1 ADM(22-52) sensitive receptors, the activation of which is likely to impair Ca2+ influx. In contrast, ADM stimulates medullary chromaffin cells to release catecholamines, which in turn enhance aldosterone secretion acting in a paracrine manner. Also this effect of ADM occurs via CGRP1 receptors, which are coupled with the adenylate cyclase-dependent cascade. There is indication that in vivo these two opposite effects of ADM on ZG may interact with each other when normal aldosterone secretion has to be restored. ADM exerts a mitogenic effect on rat ZG, acting via CGRP1 receptors that activate the tyrosine kinase-dependent mitogen-activated protein kinase cascade. These findings, along with the demonstration of a high level of ADM gene expression in adrenocortical adenomas and carcinomas, may suggest a role for ADM as adrenocortical growth stimulator and tumor promoter. PAMP, like ADM, suppresses aldosterone response of ZG cells to Ca2+-dependent agonists, but, in contrast with ADM, it inhibits catecholamine release by adrenal medulla. Both effects of PAMP are mediated by PAMP(12-20) sensitive receptors, whose signaling mechanism is likely to involve the blockade of voltage-gated Ca2+ channels. The concentrations attained by ADM and PAMP in the blood rule out the possibility that they act as true circulating hormones. Conversely, their content in the hypothalamo-pituitary complex and adrenal gland is consistent with a paracrine mechanism of action, which may play an important role in pathophysiological conditions where the function of the HPA axis has to be reset. PMID- 11407763 TI - Freeze-substitution: origins and applications. AB - Freeze-substitution is a physicochemical process in which biological specimens are immobilized and stabilized for microscopy. Water frozen within cells is replaced by organic solvents at subzero temperatures. Freeze-substitution is widely used for ultrastructural and immunocytochemical analyses of cells by transmission and scanning electron microscopy. Less well recognized is its superiority over conventional chemical fixation in preserving labile and rare tissue antigens for immunocytochemistry by light microscopy. In the postgenome era, the focus of molecular genetics will shift from analyzing DNA sequence structure to elucidating the function of gene networks, the intercellular effects of polygenetic diseases, and the conformational rearrangements of proteins in situ. Novel strategies will be needed to integrate knowledge of chemical structures of normal and abnormal macromolecules with the physiology and developmental biology of cells and tissues from whole organisms. This review summarizes the progress and future prospects of freeze-substitution for such explorations. PMID- 11407764 TI - Myosins in protists. AB - This review focuses on selected papers that illustrate an historical perspective and the current knowledge of myosin structure and function in protists. The review contains a general description of myosin structure, a phylogenetic tree of the myosin classes, and descriptions of myosin isoforms identified in protists. Each myosin is discussed within the context of the taxonomic group of the organism in which the myosin has been identified. Domain structure, cellular location, function, and regulation are described for each myosin. PMID- 11407765 TI - Orthodontics and the health care industry. PMID- 11407766 TI - Dose reduction by direct-digital cephalometric radiography. AB - Patient radiation exposure was determined for conventional and direct-digital cephalometric radiography. An anthropomorphic phantom was positioned to expose lateral cephalographs from the patient's left side. The conventional radiographs were exposed with a Siemens Orthophos C unit (77 kV, 14 mA, 0.5 s) and a film screen system of a relative speed of 400. The direct-digital radiographs were exposed with a Siemens Orthophos DS Ceph (73 kV, 15 mA, 15.8 s). A set of 108 thermoluminescence detectors (TLDs; Bicron STI/Harshaw, Solon, Ohio) was used for dose measurements. For each measurement, 84 TLDs were placed at the surface of the head and neck, as well as inside the phantom, at anatomically relevant positions. The remaining detectors were employed for calibration purposes and quality control. The highest absorbed doses were recorded for the conventional technique at the skin of the left parotid region (132 microGy), in the left parotid gland (103 microGy), and in the ocular lens of the left eye (81 microGy). Digital cephalometry resulted in an absorbed dose about 2 times lower than the dose received by the conventional technique. The effective doses had the same relation (conventional 2.3 microSv; digital 1.1 microSv). The results demonstrate that direct-digital cephalometric radiography cuts the patient's dose in half compared with the conventional screen-film technique. Direct-digital cephalometry is more advantageous than the conventional technique from the perspective of radiation protection. PMID- 11407767 TI - Orthodontic outcomes assessment using the peer assessment rating index. AB - The purpose of the present study was to gain more information on the assessment of orthodontic treatment using the PAR Index. The PAR Index was used to assess the study casts of 54 cases treated by 5 orthodontists who successfully completed Phase III of the American Board of Orthodontics. These peer-reviewed cases were considered to represent a subjective opinion of an excellent treatment standard. The PAR scores for the Board-accepted cases were then compared with the PAR scores of 51 cases consecutively treated by orthodontic graduate students. The pretreatment, posttreatment, and percent changes in PAR scores for the Board cases and the graduate student-treated cases were compared. The PAR Index scores and treatment duration were analyzed for the entire sample using general linear models techniques. Results of the study showed no statistically significant difference between the Board-accepted cases and the graduate student-treated cases for any single component of the PAR Index at either the pretreatment or posttreatment times. However, the percent reduction in the mean PAR score for the Board-accepted cases was significantly more than the reduction for the graduate student-treated cases (87.9%+/-9.5% vs 81.7%+/-15.3%). Analysis of treatment duration showed that a higher pretreatment PAR score and a greater percent reduction in PAR score were significantly associated with longer treatment durations. PMID- 11407768 TI - Skeletal changes of Herbst appliance therapy investigated with more conventional cephalometrics and European norms. AB - We measured the skeletal effects of Herbst appliance treatment in a retrospective sample of 25 boys (aged 10.7-14.5 years) and 25 girls (aged 10.7-14.3 years). Selection criteria were (1) a pretreatment full Class II molar relationship (ANB angle: average, 6.7 degrees; range, 2.5-10.5 degrees) and (2) a posttreatment full Class I or overcorrected Class I molar relationship within 6-8 months. A first t-test was used to evaluate variations between pre- and posttreatment cephalometric measures. Then, compared with the appropriate age- and sex-matched European norm, every pre- and posttreatment value was transformed into a z-score on the distribution of the norm value and a second t-test was performed. The second t-test was to study variations between pre- and posttreatment z-scores in order to neutralize the effect of natural growth. Posttreatment, the mandible showed a remarkable forward repositioning without opening of the gonial angle, particularly in males. Only ANB and Xi-CF-PTV angles were significantly different when the effect of normal growth was excluded. In males, ramus height and mandibular basal length were significantly increased when total variation was considered (ie, not excluding the effect of normal growth). In females, only the mandibular ramus height was significantly increased. In conclusion, even short term Herbst therapy can be efficacious, with the most frequent effect being mandibular forward repositioning followed by mandibular ramus elongation. The statistical procedure used counteracts the effect of growth and sex on the results. Moreover, z-scores are adimensional measures with which any kind of parameter may be compared and scaled to each other in the perspective of a more reliable mutivariated interpretation of cephalometric variables. PMID- 11407769 TI - An electromyographic investigation of the first six months of progressive mandibular advancement of the Herbst appliance in adolescents. AB - The effects of the progressive activation of the Herbst appliance on the activity of the masseter and temporalis elevator muscles of the mandible were monitored in a group of 14 consecutively treated 10-15-year-old subjects with an Angle Class II, division I malocclusion. A cast silver splint Herbst appliance was activated in multiple stages at a rate of 2 mm/2 mo. The functionality of the superficial masseter and anterior portion of the temporalis muscles was monitored at maximum bite force using surface electromyography (EMG). The EMG recordings were taken at an incisal edge-to-edge position and a retruded mandibular position, both at a vertical interincisal separation of 3 mm using an acrylic bite plate. Measurements of maximum voluntary isometric clenches were taken during adaptive functional changes at pretreatment (baseline) and during the first 6 months of Herbst appliance therapy. Results showed great individual and inter- and intrasessional differences in electromyographic activity of the muscles before and during treatment. At the retruded position, the masseteric activity increased by the sixth month while temporalis activity remained at the same level. Following treatment, the masseteric imbalance was reduced, but the temporalis imbalance was unchanged. At the edge-to-edge position, masseteric activity increased by the sixth month, while temporalis activity remained unchanged. The masseteric imbalance was reduced by the sixth month, while the temporalis imbalance was reduced from the fourth month into treatment. The results imply a favorable muscular response to a progressive regime of Herbst appliance activation. PMID- 11407770 TI - Assessment of root resorption and root shape: periapical vs panoramic films. AB - A radiographic examination is an essential part of the diagnostic process in orthodontics. However, what radiographs are needed to properly evaluate root shape and position? Most clinicians order panoramic or periapical radiographs in addition to the cephalometric radiograph. The purpose of this study was to find out whether one type of film is more accurate than the other in the pretreatment evaluation of root shape and the posttreatment computation of apical root resorption. Pretreatment and posttreatment panoramic films and full-mouth periapical films from 42 patients who completed fixed orthodontic treatment were assessed for tooth length and root shape. Panoramic films showed significantly greater average apicaL root resorption than periapical films for the 743 teeth surveyed. The greatest differences were found in the lower incisors, the least in the maxillary incisors. Classification of root shape was significantly different between the 2 types of radiographs. Root dilacerations and other abnormal shapes, clearly visible on periapical films, often appeared normal on panoramic films. The findings strongly suggest that root shape is much harder to assess on panoramic films. We conclude that, in cases where the apices are obscured or other factors are present that might suggest higher risk for root resorption or vertical bone loss, periapical films should be ordered. The use of panoramic films to measure pre- and posttreatment root resorption may overestimate the amount of root loss by 20% or more. PMID- 11407771 TI - Investigation of bacteremia after orthodontic banding and debanding following chlorhexidine mouth wash application. AB - This study investigates the prevalence of bacteremia after orthodontic banding and debanding, following the application of a 0.2% chlorhexidine gluconate mouthwash. The banding and debanding groups were each composed of 40 young adult patients. In the banding group, patients were asked to rinse their mouth with chlorhexidine gluconate for 60 seconds just prior to fitting of the bands. In the debanding group, they were asked to use the mouthwash immediately before removal of bands and brackets. In both groups pre- and post-treatment blood samples were obtained with a strict aseptic technique. In the banding group, no bacteremia was detected in the pretreatment sample and 2.5% post-treatment bacteremia was detected in the post-treatment sample. In the debanding group, 2.5% bacteremia was found in both the pre- and post-treatment samples. The prevalence of post treatment bacteremia found in the present study were compared with the findings of 2 preliminary studies in which the prevalence of bacteremia had been investigated after banding and debanding without a prior application of chlorhexidine mouthwash. The application of chlorhexidine mouthwash resulted in a decrease in the prevalence of bacteremia after banding and debanding, but the decrease was not statistically significant. PMID- 11407772 TI - A comparative study of Caucasian and Japanese mandibular clinical arch forms. AB - The purpose of this study was to clarify morphological differences between Caucasian and Japanese mandibular clinical arch forms in Class I, II, and III malocclusions. The study included 60 Class I, 50 Class II, and 50 Class III cases from each ethnic group. The most facial portion of 13 proximal contact areas was digitized from photocopied images of the mandibular dental arches. Clinical bracket points were calculated for each tooth based on mandibular tooth thickness data. Four linear and 2 proportional measurements were taken. The dental arches were classified into square, ovoid, and tapered forms to determine and compare the frequency distributions between the 2 ethnic groups. The Caucasian population had a statistically significant decreased arch width and increased arch depth compared with the Japanese population. When the subjects were regrouped by arch form, no statistically significant difference in arch dimension was observed between the 2 ethnic groups in any of the arch form samples. Our results suggest that there is no single arch form specific to any of the Angle classifications or ethnic groups. It appears to be the frequency of a particular arch form that varies among Angle classifications or ethnic groups. PMID- 11407773 TI - Modulation of the stretch reflex of jaw-closing muscles in different modes and phases of respiration. AB - The objective of this study was to investigate whether and how changes in the mode of respiration affect the electromyographic activity of human jaw-closing muscles. Fifteen men were examined in this study. A pair of surface electrodes was attached bilaterally to the masseter and anterior and posterior temporalis muscles for electromyographic recording. Respiratory movements of the chest wall and nasal airflow were recorded simultaneously. Recordings were performed with subjects in the sitting position during quiet nasal and oral respiration. The stretch reflex of jaw-closing muscles was elicited by randomly tapping the chin with an impulse hammer. In 11 subjects, we measured nasal resistance with a rhinomanometer. The amplitude of electromyographic activities of the masseter and anterior temporalis muscles during oral respiration was significantly less than that during nasal respiration, whereas that of the posterior temporalis muscle showed no significant difference between the different modes of respiration. Furthermore, the reduction in the amplitude of the electromyographic activity was more evident in the inspiratory phase during oral respiration. There was a significant positive correlation between the ratio of the reflex amplitude during inspiration in the 2 respiratory modes and nasal resistance for the masseter muscle, but not for the anterior temporalis muscle. These results suggest that the reflexive electromyographic activity of some human jaw-closing muscles is modulated during oral respiration. PMID- 11407774 TI - Measurements of mandibular length: a comparison of articulare vs condylion. AB - This study examines the validity of articulare for mandibular length measurements by exposing 3 lateral cephalograms on each of 60 consecutive patients. The radiographs were exposed with the patient in a closed-mouth position in habitual occlusion, a closed-mouth position with the patient in centric relation, and in an open-mouth position. The linear distances (mm) of articulare (Ar) to pogonion (Pog), Ar to gonion (Go), and Go to Pog were measured on the 2 closed-mouth cephalograms and compared with each other as well as the linear distances of condyle (Co) to Pog, Co to Go, and Go to Pog measured from the open-mouth cephalogram on each individual. Product-moment correlation coefficients were used to measure the linear associations among the mandibular measurements from the 3 techniques. Repeated measures analyses of variance were also fit to estimate the correlations between the 3 measurements adjusted for age and sex. The results of this study show that measurements taken from both closed-mouthed techniques agreed extremely well (intraclass correlation coefficient = 0.99). In addition, measurements from both closed-mouth techniques highly agreed with the corresponding measurements taken with the open-mouth technique (intraclass correlation coefficient = 0.94). This data suggests that measurements taken from Ar correlate very well with measurements taken from Co and that this correlation is not dependent on whether the patient is positioned in habitual occlusion or centric relation. PMID- 11407775 TI - Factors influencing the predictability of soft tissue profile changes following mandibular setback surgery. AB - The objective of this cephalometric study was to assess long-term changes in the soft tissue profile following mandibular setback surgery and investigate the presence of factors that may influence the soft tissue response to skeletal repositioning. The subjects enrolled were 80 consecutive mandibular prognathism patients operated with bilateral sagittal split osteotomy and rigid fixation. Lateral cephalograms were taken at 6 occasions: immediate presurgical, immediate postsurgical, 2 and 6 months postsurgical, and 1 and 3 years postsurgical. The subjects were grouped according to gender and magnitude of setback. Ratios of soft tissue to hard tissue movements were calculated for the subgroups. Females generally demonstrated greater ratios than males with a statistically significant difference for the upper lip and chin (P < .05). Postsurgical alterations in the profiles were more predictable in patients with larger setbacks compared to patients with smaller ones. Skeletal relapse had a profound influence on long term profile changes. Based on these findings, it is proposed that the database used in prediction software be adjusted to account for such factors in an attempt to improve the accuracy of computerized treatment simulations. PMID- 11407776 TI - Severe dental open bite malocclusion with tongue reduction after orthodontic treatment. AB - We treated a 21-year-old woman with a severe open bite and macroglossia with a standard edgewise appliance and without partial glossectomy. This was followed by retention using a Begg-type plate retainer for the upper dental arch and a fixed canine-to-canine for the lower arch. A crib was added to the upper plate retainer for suppression of a tongue thrust. The lower arch relapsed during the retention period, with a widening of the intermolar distance, flaring of the anterior teeth, and increased mobility of the teeth. We chose tongue reduction to resolve these problems and one-third of the middle dorsal part of the tongue was excised. After the tongue reduction, the patient experienced no functional problem in mastication, swallowing, and gustation, but she complained of mild speech difficulty and slight pain on the dorsal portion of her tongue. These symptoms disappeared 6 months after surgery. At this time, the mandibular dental arch was markedly improved. The flared lower dental arch had returned to an upright position and the tooth mobility reduced to normal. No appliance was used after surgery. Most of the recovery changes occurred within 4 months. This case highlights the importance of the teeth tending to move toward a balance between the tongue pressure from the inside and labio-buccal pressure from the outside. PMID- 11407777 TI - Re: A plea for agreement. PMID- 11407778 TI - On telling it like it was. PMID- 11407779 TI - SCN efferents to peripheral tissues: implications for biological rhythms. AB - The suprachiasmatic nucleus (SCN) is the principal generator of circadian rhythms and is part of an entrainment system that synchronizes the animal with its environment. Here, we review the possible communication of timing information from the SCN to peripheral tissues involved in regulating fundamental physiological functions as revealed using a viral, transneuronal tract tracer, the pseudorabies virus (PRV). The sympathetic nervous system innervation of the pineal gland and the sympathetic outflow from brain to white adipose tissue were the first demonstrations of SCN-peripheral tissue connections. The inclusion of the SCN as part of these and other circuits was the result of lengthened postviral injection times compared with those used previously. Subsequently, the SCN has been found to be part of the sympathetic outflow from the brain to brown adipose tissue, thyroid gland, kidney, bladder, spleen, adrenal medulla, and perhaps the adrenal cortex. The SCN also is involved in the parasympathetic nervous system innervation of the thyroid, liver, pancreas, and submandibular gland. Individual SCN neurons appear connected to more than one autonomic circuit involving both sympathetic and parasympathetic innervation of a single tissue, or sympathetic innervation of two different peripheral tissues. Collectively, the results of these PRV studies require an expansion of the traditional roles of the SCN to include the autonomic innervation of peripheral tissues and perhaps the modulation of neuroendocrine systems traditionally thought to be controlled solely by hypothalamic stimulating/inhibiting factors. PMID- 11407780 TI - Circadian photoreception in Drosophila: functions of cryptochrome in peripheral and central clocks. AB - In Drosophila melanogaster, disruption of night by even short light exposures results in degradation of the clock protein TIMELESS (TIM), leading to shifts in the fly molecular and behavioral rhythms. Several lines of evidence indicate that light entrainment of the brain clock involves the blue-light photoreceptor cryptochrome (CRY). In cryptochrome-depleted Drosophila (cry(b)), the entrainment of the brain clock by short light pulses is impaired but the clock is still entrainable by light-dark cycles, probably due to light input from the visual system. Whether cryptochrome and visual transduction pathways play a role in entrainment of noninnervated, directly photosensitive peripheral clocks is not known and the subject of this study. The authors monitored levels of the clock protein TIM in the lateral neurons (LNs) of larval brains and in the renal Malpighian tubules (MTs) of flies mutant for the cryptochrome gene (cry(b)) and in mutants that lack signaling from the visual photopigments (norpA(P41)). In cry(b) flies, light applied during the dark period failed to induce degradation of TIM both in MTs and in LNs, yet attenuated cycling of TIM was observed in both tissues in LD. This cycling was abolished in LNs, but persisted in MTs, of norpA(P41);cry(b) double mutants. Furthermore, the activity of the tim gene in the MTs of cry(b) flies, reported by luciferase, seemed stimulated by lights-on and suppressed by lights-off, suggesting that the absence of functional cryptochrome uncovered an additional light-sensitive pathway synchronizing the expression of TIM in this tissue. In constant darkness, cycling of TIM was abolished in MTs; however, it persisted in LNs of cry(b) flies. The authors conclude that cryptochrome is involved in TIM-mediated entrainment of both central LN and peripheral MT clocks. Cryptochrome is also an indispensable component of the endogenous clock mechanism in the examined peripheral tissue, but not in the brain. Thus, although neural and epithelial cells share the core clock mechanism, some clock components and light-entrainment pathways appear to have tissue-specific roles. PMID- 11407781 TI - Postmortem stability of melatonin receptor binding and clock-relevant mRNAs in mouse suprachiasmatic nucleus. AB - The stability of receptor proteins and mRNAs in brain tissue is variable after death. As a prelude to quantitative studies of melatonin receptor density and clock gene expression in the human brain, the stability of these macromolecules was examined in the mouse brain under simulated postmortem conditions using the model of Spokes and Koch. In the mouse suprachiasmatic nucleus (SCN), melatonin receptor binding was significantly reduced after 18 to 24 h under postmortem conditions. Two mRNAs that are rhythmically expressed in the SCN, mPer1 and prepropressophysin (AVP), also decreased significantly over the interval studied, and mPer1 declined more rapidly than AVP. Both mPer1 and AVP mRNA levels in the SCN declined more rapidly in vivo than under postmortem conditions, suggesting that the degradation of these mRNAs is an active process. The results indicate that quantitative studies of melatonin receptor density on human postmortem material are feasible and that detection of rhythmic gene expression in the human SCN will likely require collection of specimens with a rather short (< 8 h) interval from death to tissue collection. The relative stability of melatonin receptor binding in the SCN also suggests that receptor binding may be a reliable marker for the location of the SCN in studies assessing clock gene expression in postmortem material. PMID- 11407782 TI - Photoperiod modulates the inhibitory effect of in vitro melatonin on lymphocyte proliferation in female Siberian hamsters. AB - In Siberian hamsters (Phodopus sungorus), short days suppress reproductive function and lymphocyte proliferation. To determine whether melatonin influences cell-mediated immunity through a direct action on lymphocyte proliferation, in vitro responsiveness to mitogens and melatonin was assessed in systemic and splenic lymphocytes from adult female Siberian hamsters housed in either long or short days for 13 weeks. Short days provoked reproductive regression and reduced lymphocyte proliferation. Physiological concentrations of melatonin (50 pg/ml) inhibited in vitro proliferation of circulating lymphocytes, whereas higher concentrations (> or = 500 pg/ml) were required to inhibit proliferation of splenic lymphocytes. Immunomodulatory effects of melatonin were restricted to lymphocytes from long-day hamsters-in vitro melatonin had no effect on circulating or splenic lymphocytes from females in short days. Responsiveness to melatonin in short-day lymphocytes may be restrained by the already expanded nightly pattern of melatonin secretion in short days. These data support the hypothesis that melatonin acts directly on lymphocytes from long-day hamsters to suppress blastogenesis. PMID- 11407783 TI - Phase response relationships between light pulses and the melatonin rhythm in rats. AB - There is some controversy whether phase response curves constructed from studies conducted after acute release into constant darkness (Type II) or after prolonged constant darkness are comparable. This study investigated the effects of brief low-intensity light pulses on the onset of 6-sulphatoxymelatonin excretion in rats 48 to 60 h after lights-off and after 14 days of continuous darkness. In the former condition, maximum phase delays occurred between 4 and 6 h after expected lights-off, but no phase advances were observed within 2 days of the presentation of the stimulus. When the times of the pulses were plotted in relation to the individual onsets, peak light-induced phase delays occurred 0 to 2 h after melatonin onset. After 14 days in continuous darkness, the peak phase delays also occurred 0 to 2 h after melatonin onset and were slightly but significantly smaller. No significant phase advances were observed. In a separate small series of experiments, the temperature rhythm of rats was shown to be delayed by a comparable degree to that of melatonin by light pulses 2 and 4 h after expected lights-off under the Type II conditions. It is concluded that phase response curves conducted under Type I and Type II conditions are comparable. PMID- 11407784 TI - Aging, reproduction, and the melatonin rhythm in the Siberian hamster. AB - The present study tested the hypothesis that responsiveness to melatonin, the presence of the melatonin rhythm in circulation, and parameters of the GnRH neuron system are sustained across the aging continuum in Siberian hamsters. Afternoon melatonin injections induced testicular atrophy in 42% of aged males compared with 100% of adult males. The proportion of aged males failing to respond to the melatonin injections was similar to the proportion that failed to undergo testicular regression upon exposure to short days. Exposure to short days induced testicular atrophy in juvenile and adult hamsters; however, regression was incomplete or absent in 43% of aged males. The nocturnal rise in melatonin was similar with regard to duration and peak amplitude, and appropriate with respect to photoperiod in 25-day-old juveniles, adult (5 months), and aged (17 months) hamsters. Neither advanced age nor timed melatonin treatments affected GnRH neuron numbers or distribution. Fertility was maintained in aged and adult males to a comparable extent with respect to latency to first litter and number of pups per litter; reproductive success was dramatically reduced in aged compared with adult females. Because melatonin rhythms accurately reflect day length information throughout the continuum from puberty to advanced age, the present evidence suggests that limitations in testis regression in response to short days or exogenous melatonin in a subset of aged males result from a reduced ability to respond to melatonin. In the wild, failure to undergo testicular regression in the presence of shortening day lengths may extend the breeding season of aged males. PMID- 11407785 TI - Seasonal variation in endogenous serum melatonin profiles in goats: a difference between spring and fall? AB - The pineal hormone melatonin serves as a signal of day length in the regulation of annual rhythms of physiological functions and behavior. The duration of high melatonin levels in body fluids is proportional to the duration of the dark period of the day. Due to the direct suppression of melatonin by light, the overt melatonin rhythm may differ from the endogenous rhythm driven by the hypothalamic circadian clock. The aim of this study was to find out possible differences between the overt and endogenous melatonin rhythms in goats during the course of a year. Seven Finnish landrace goats (nonlactating females) were kept under artificial lighting that approximately simulated the annual changes of day length at 60 degrees N. Blood samples for melatonin measurements by radioimmunoassay were collected at 2-h intervals during six seasons: winter (light:dark 6:18 h), early spring (10:14), late spring (14:10), summer (18:6), early fall (14:10), and late fall (10:14). Melatonin profiles were determined for 2 consecutive days, first in light-dark (LD) conditions and then in continuous darkness (DD). In LD conditions, the profiles matched the dark period with one exception: In winter, the mean peak duration was significantly shorter than the scotoperiod. In DD conditions, two types of endogenous melatonin patterns were found: a "winter pattern" (peak duration 13-15 h) in winter, early spring, early fall, and late fall, and a "summer pattern" (duration about 11 h) in late spring and summer. Thus, in equal habitual LD conditions in late spring and early fall (LD 14:10), the endogenous melatonin rhythms were not quite similar: The pattern in late spring resembled that in summer, and the pattern in early fall that in winter. These results suggest that, in addition to the light-adjusted overt melatonin rhythm, the endogenous rhythm of melatonin secretion varies during the course of a year. PMID- 11407786 TI - Hibernation effects on memory in European ground squirrels (Spermophilus citellus). AB - Effects of hibernation on memory were tested in European ground squirrels (Spermophilus citellus). The animals were trained in summer to successfully accomplish two tasks: a spatial memory task in a maze and an operant task on a feeding machine. One group hibernated normally, and the other was prevented from hibernation by maintaining ambient temperature at 22 degrees C. In spring, the same tasks were repeated for both groups and their individual performances compared to the initial training phase. The experimental groups differed significantly in both tasks. The nonhibernating animals had higher levels of retention and needed significantly fewer trials to relearn the tasks than the group that had hibernated. In addition to testing the retention of conditioned tasks, social memory was also studied. The ground squirrels were given a social recognition test in spring with one familiar and one unfamiliar conspecific. In contrast to the conditioned tasks, social memory did not seem to be affected by hibernation. The results indicate negative effects of hibernation on the retention of conditioned tasks, which could produce important constraints on animals. A potential explanation for this memory loss might be changes in neuronal activity, which occur during hibernation. PMID- 11407787 TI - Postprandial triacylglycerol responses in simulated night and day shift: gender differences. AB - A number of reports suggest that shift workers have an increased risk of coronary heart disease (CHD). One contributing factor may be the consumption of meals at night with consequent altered postprandial responses. This study investigated circulating triacylglycerol (TAG), a possible risk factor for CHD, after meals during a simulated day and night shift. Twenty-five healthy participants (10 women and 15 men) were studied. They were given a pre-meal at 0800 h and a test meal at 1330 h on a simulated day shift and then an identical pre-meal at 2000 h and test meal at 0130 h, respectively, on a simulated night shift with maintained wakefulness. Blood was sampled for 9 h after the test meal for analysis of basal and postprandial plasma TAG levels. ANOVA for repeated measures indicated higher TAG in men compared with women (p < 0.0001) and higher responses at night in both genders (p = 0.027). Incremental area under the curve (IAUC) analysis indicated that men had significantly increased postprandial TAG levels at night compared with the day: (IAUC 0-540 min, mean +/- SEM) 253.29 +/- 28.73 versus 148.33 +/- 17.28 mmol/L x min, respectively, p = 0.025. In women, night and day responses (61.16 +/- 8.93 versus 34.09 +/- 7.87 mmol/L x min, respectively, p = 0.457) were not significantly different. Circulating TAG remained elevated for longer at night in the men compared with the women (p = 0.009). This study demonstrates the existence of gender and time-of-day differences in TAG responses to a meal. These raised TAG levels at night, for a prolonged time in men, may be relevant to the increased risk of CHD in shift workers. PMID- 11407788 TI - The effect of osteogenic protein-1 on the healing of segmental bone defects treated with autograft or allograft bone. AB - BACKGROUND: Large amounts of bone graft are frequently used to elicit the healing of bone defects resulting from reconstructive procedures. Autograft and allograft bone are often used, but each has its limitations. Bone morphogenetic proteins (BMPs) improve the healing of segmental bone defects treated with autograft or allograft. The objective of the present study was to determine the effect of implantation of a recombinant osteogenic protein-1 (OP-1) in combination with bone graft on the healing of a critical-sized (2.5-cm) segmental defect in canine ulnae. METHODS: Either autograft bone, allograft bone, osteogenic protein-1 (OP 1) mixed with type-1 bovine collagen, or various combinations of OP-1 and collagen (OP-1 device) mixed with allograft or autograft were implanted in the segmental bone defects. The combinations included 67% bone graft with 33% OP-1 device and 33% bone graft with 67% OP-1 device. The healing of the defects was assessed with radiographic, biomechanical, and histological studies. The animals were killed at twelve weeks postoperatively. RESULTS: The use of the OP-1 device alone or any combination of autograft or allograft bone and the OP-1 device demonstrated improved healing on radiographic, mechanical, and histological studies compared with that demonstrated after use of autograft or allograft bone alone. The highest radiographic and histological grades and the greatest mechanical strength were achieved with the use of 33% allograft and 67% OP-1 device, although no significant differences were observed among the different groups containing the OP-1 device. At twelve weeks postoperatively, the defects treated with any amount of the OP-1 device obtained greater mechanical strength than that obtained by autograft bone alone. CONCLUSIONS: Major bone defects may be treated with allograft bone combined with the OP-1 device, instead of autograft alone, to avoid complications associated with the use of autograft. The combination of allograft bone and the OP-1 device resulted in optimum healing of the defect, according to the radiographic, mechanical, and histological parameters measured in this study. CLINICAL RELEVANCE: The combination of freeze dried allograft bone with the OP-1 device is an attractive graft material for the treatment of large bone defects. Although similar results were observed when autogenous bone graft was used in combination with the OP-1 device, the results of the present study suggest that allograft, because of its relatively unlimited supply, can be substituted without reduced efficacy. In addition, avoiding the need to harvest autogenous bone eliminates the additional operative time and risk associated with a second surgical procedure. PMID- 11407789 TI - Hydroxyapatite-coated acetabular components. Histological and histomorphometric analysis of six cups retrieved at autopsy between three and seven years after successful implantation. AB - BACKGROUND: Important questions remain regarding the use of hydroxyapatite-coated acetabular components in total hip arthroplasty. What is the relation of resorption of the hydroxyapatite coating to enduring fixation? Will unresorbed or dislodged hydroxyapatite particles cause adverse tissue reactions? Retrieval studies of clinically well-functioning acetabular components should help to answer these questions. METHODS: We examined six clinically successful hydroxyapatite-coated cementless acetabular components that were retrieved at autopsy between 3.3 and 6.6 years after implantation. All components were of the same design. The prostheses and the surrounding bone were prepared for qualitative histological and quantitative histomorphometric analysis. The percentage of bone growth onto the implant, the relative bone area around the implant, the extent of residual hydroxyapatite coating, and the coating thickness were measured. RESULTS: All of the cups showed bone ongrowth, with a mean bone implant contact (and standard deviation) of 36.5% +/- 13.5%. The contact area was the same in all three zones delineated by DeLee and Charnley. The extent and thickness of the hydroxyapatite layer were much reduced in the specimens from older patients and in those associated with a longer duration of implantation. Degradation of the hydroxyapatite coating by osteoclasts was observed. We did not observe loose hydroxyapatite granules far from the coating, nor did we note any adverse tissue reaction to these granules. In contrast, polyethylene debris was noted in approximately half of the empty screw-holes. CONCLUSIONS: Cell-mediated hydroxyapatite resorption seems to be the main reason for loss of hydroxyapatite coating. The area of bone ongrowth was within a certain range (20% to 50%) of the measured surfaces, and it was independent of the amount of hydroxyapatite residue. The hydroxyapatite coating showed a slow rate of resorption with time, without any adverse tissue reactions. PMID- 11407790 TI - Immunolocalization of matrix metalloproteinases in partial-thickness defects in pig articular cartilage. A preliminary report. AB - BACKGROUND: Partial-thickness defects in mature articular cartilage do not heal spontaneously. Attempts at repair often result in limited integration between the repair tissue and the surrounding cartilage, with formation of chondrocyte clusters adjacent to a zone of cartilage necrosis. In wound repair, spatially and temporally controlled expression of matrix metalloproteinases and their inhibitors have been implicated in proteolytic degradation of damaged extracellular matrix components, but the sequence of events following damage to cartilage is unknown. To determine this sequence, we studied the distribution of matrix metalloproteinases and their inhibitors during early in vivo repair of partial-thickness defects in pig articular cartilage. METHODS: With use of a model that elicits the ingrowth of mesenchymal cells into partial-thickness defects, partial-thickness defects were created in knee joint cartilage. The distributions of matrix metalloproteinase-1, 2, 3, 9, 13, and 14; tissue inhibitors of metalloproteinase-1 and 2; and the neoepitope DIPEN341 specifically generated following matrix metalloproteinase cleavage of aggrecan were determined by immunolocalization of repair tissue and surrounding cartilage excised from immature pigs during the first eight weeks of repair and from adult minipigs at eight days and three weeks. RESULTS: Synthesis of matrix metalloproteinase-13 was usually confined to hypertrophic chondrocytes in immature cartilage and to the radial zone in adult cartilage. Following injury, strong induction of matrix metalloproteinase-13 synthesis was observed in chondrocyte clusters surrounding lesions in all of the animals. The migration of macrophages into defects was prominent at two and eight days, with synthesis and deposition of matrix metalloproteinase-9 onto damaged cartilage matrix and newly synthesized matrix in the defect. The DIPEN341 neoepitope was localized to damaged cartilage matrix at eight days and six weeks, indicating partial degradation of aggrecan. Focal synthesis of matrix metalloproteinase-1, 3, and 14 and of tissue inhibitor of metalloproteinase-1 occurred at later times, suggesting continuous remodeling of the increasingly compact repair tissue. CONCLUSIONS: The expression of matrix metalloproteinase-13 by normal hypertrophic chondrocytes and the induction of synthesis in chondrocyte clusters adjacent to the zone of cartilage necrosis suggest that this enzyme participates in the pericellular proteolysis required for lacunar expansion. The localization of matrix metalloproteinase-9 to damaged cartilage matrix suggested matrix proteolysis, which was confirmed with DIPEN341 localization. Reduced matrix metachromasia persisted and was colocalized with DIPEN341 at six weeks. However, under the conditions investigated, there was only limited proteolytic degradation in the zone of cartilage necrosis. This may render the zone mechanically weakened, thereby contributing to subsequent instability of the region, and may form a barrier to integration of repair tissue with viable cartilage. CLINICAL RELEVANCE: Osteoarthritis initially involves the superficial layers of cartilage. The development of procedures to promote the healing or repair of early defects will have major advantages in terms of disease alleviation as well as economic importance. Identification of the enzymes involved in the early repair of partial-thickness defects in articular cartilage is clinically relevant because proteolysis of damaged matrix has to take place in order for repair tissue to integrate with surrounding healthy cartilage. PMID- 11407791 TI - Neuropathic arthropathy of the elbow. A report of five cases. AB - BACKGROUND: Neuropathic arthropathy of the elbow is rare and characterized by a painless but unstable articulation. The functional capacity of patients with this condition has not been reviewed in detail. METHODS: Five male patients, with an average age of fifty-one years, were treated for neuropathic arthropathy of the elbow. The underlying conditions associated with the arthropathy included syringomyelia, insulin-dependent diabetes mellitus, end-stage renal failure, and two cases of polyneuropathy of unknown cause. Four patients sought medical attention after a specific traumatic event. Peripheral sensory and motor dysfunction was present in each patient. Radiographs of the elbow revealed dislocation, fracture fragmentation, and heterotopic ossification. Our management of the neuropathic elbows centered on maintenance of a functional arc of motion through physical therapy aimed at regaining muscle strength and the use of orthoses for support. Operative treatment was performed for an associated ulnar or radial nerve compression syndrome in three patients, and an open reduction and internal fixation of an unstable proximal ulnar nonunion associated with loose implants was performed in one. RESULTS: The patients were followed for an average of nineteen months, with a range of twelve to thirty-six months. All patients had a pain-free elbow with a functional range of motion at the most recent follow-up examination, and none wished to have further treatment. The operatively treated ulnar nonunion united successfully. All three patients treated surgically for an associated nerve compression syndrome had recovery of nerve function. CONCLUSION: In the face of instability and gross distortion of the joint, the patients in this series demonstrated remarkably good function. PMID- 11407792 TI - Dega osteotomy for the treatment of congenital dysplasia of the hip. AB - BACKGROUND: In 1969 Dega described a transiliac osteotomy to treat residual acetabular dysplasia secondary to congenital hip dysplasia or dislocation. We were unable to find a thorough description of the technique in the English language orthopaedic literature, and the number of clinical follow-up series is small. METHODS: Twenty-two children (twenty-four hips) with an average age of five years and ten months and varying degrees of congenital hip dysplasia, subluxation, or dislocation were treated with a Dega osteotomy. Twenty hips (83%) had a concomitant femoral osteotomy and thirteen (54%) had an anterior open reduction of the hip in addition to the Dega osteotomy. To be included in the study group, each patient had to have complete clinical documentation of the range of motion, presence or absence of a limp, limb-length discrepancy, hip pain, and limitation of activity. Radiographs were reviewed to determine the acetabular index, the center-edge angle, whether the Shenton line was intact or broken, and any change in the projection of the obturator foramen. RESULTS: At an average of fifty-five months postoperatively, all patients demonstrated unlimited physical activity and no limp. The average acetabular index changed from 33 degrees preoperatively to 12 degrees at the time of follow-up. The center-edge angle ranged from less than -30 degrees to 18 degrees preoperatively and from 18 degrees to 40 degrees (average, 31 degrees) at the time of follow-up. A change in the obturator foramen was observed in fourteen hips (58%). The Shenton line was broken in seventeen hips preoperatively but in none postoperatively. One Dega osteotomy was revised immediately after the index operation, and three hips underwent late repeat correction of the proximal part of the femur; one of the repeat corrections was performed together with a repeat Dega osteotomy. CONCLUSIONS: Our initial experience with the Dega osteotomy demonstrated it to be a valuable surgical treatment of congenital dysplasia of the hip in a child of walking age. Our experience is comparable with that of many European authors, including those reporting studies from Dega's own institution. PMID- 11407793 TI - Synergy between Staphylococcus aureus and Pseudomonas aeruginosa in a rat model of complex orthopaedic wounds. AB - BACKGROUND: We observed an interaction in animals inoculated concomitantly with Staphylococcus aureus and Pseudomonas aeruginosa during a study of the efficacy of surfactants for disinfection of orthopaedic wounds. This led us to investigate whether synergy could be demonstrated between Staphylococcus aureus and Pseudomonas aeruginosa in a rat model of complex orthopaedic wounds. METHODS: A wire was implanted into the spinous process of a lumbar vertebra of Sprague Dawley rats through a dorsal incision. Animals were divided into two groups: group one was inoculated with either Staphylococcus aureus or Pseudomonas aeruginosa, and group two received a polymicrobial inoculation with both test organisms in varying concentrations. After inoculation, the wounds were irrigated and closed. On postoperative day 14, all animals were killed and specimens from the wounds were cultured. The number of colony-forming units (CFU) of Staphylococcus aureus or Pseudomonas aeruginosa needed to cause infection in 50% of the animals (ID50) was determined with use of the Reed-Muench method. The infection rate associated with each inoculum combination was calculated, and the two groups were compared. RESULTS: The ID50 was 2.8 x 10(4) CFU for Staphylococcus aureus and 4.8 x 10(5) CFU for Pseudomonas aeruginosa. The combination of 10(3) CFU of Staphylococcus aureus with low concentrations (10(2), 10(3), or 10(4) CFU) of Pseudomonas aeruginosa yielded infection rates that were higher than those found with either organism alone at the same concentrations. The combination of 10(3) CFU of Staphylococcus aureus and 10(3) CFU of Pseudomonas aeruginosa yielded a 75% infection rate, which was significantly higher (p = 0.004) than that associated with 10(3) CFU of either organism alone. As the Pseudomonas aeruginosa concentration was increased (to 10(5), 10(6), and 10(7) CFU), this trend reversed, and the infection rate decreased to 33% (p = 0.004). Low concentrations of Pseudomonas aeruginosa (0 to 10(5) CFU) combined with 10(6) CFU of Staphylococcus aureus yielded infection rates ranging from 83% to 100%. At the higher concentrations of Pseudomonas aeruginosa (10(6) and 10(7) CFU), however, the infection rate again decreased, to 33% (p = 0.005). Only Staphylococcus aureus was isolated from the cultures of the specimens from the animals that had received a polymicrobial inoculum. CONCLUSIONS: Synergy between Staphylococcus aureus and Pseudomonas aeruginosa was demonstrated when low levels of each organism were present in the wound. As the Pseudomonas aeruginosa concentration was increased, the infection rates fell well below what would be anticipated, suggesting that low concentrations of Pseudomonas aeruginosa enhance the ability of Staphylococcus aureus to cause infection in this orthopaedic wound model. At the same time, the presence of Staphylococcus aureus in the ratios tested decreased the rate of infection by Pseudomonas aeruginosa. CLINICAL RELEVANCE: Staphylococcus aureus is a pathogen commonly seen in orthopaedic patients. The pathogenicity of Staphylococcus aureus was shown to be increased in the presence of anaerobic bacteria. This study is the first one that we are aware of that demonstrated synergy between Staphylococcus aureus and Pseudomonas aeruginosa, at low concentrations, in a wound model while at the same time showing that Staphylococcus aureus lowers the rate of Pseudomonas aeruginosa infection. PMID- 11407794 TI - Allogeneic cancellous bone graft and a Burch-Schneider ring for acetabular reconstruction in revision hip arthroplasty. AB - BACKGROUND: There is an ever-increasing number of failed hip arthroplasties associated with massive deficiency of acetabular bone stock consisting of a segmental or cavitary defect. This study was undertaken to evaluate the long-term results after use of morselized cryopreserved allogeneic bone graft and an antiprotrusio cage to treat such a deficiency. METHODS: From January 1, 1988, to January 1, 1994, forty-one patients (forty-one hips) with an acetabular defect classified as type IIl or IV according to the American Academy of Orthopaedic Surgeons system were operated on with use of a Burch-Schneider ring and morselized cryopreserved allogeneic cancellous bone graft. Thirty-eight patients (thirty-eight hips) were available for clinical and radiographic follow-up examinations at an average of 7.3 years (range, 4.2 to 9.4 years) after surgery. RESULTS: All measured clinical parameters had improved significantly by the time of the follow-up examination (p < 0.0001). Radiographs confirmed that none of the thirty-eight hips had any measurable migration or displacement of the acetabular component and that osseous consolidation occurred only within the grafted area in all patients. CONCLUSION: Acetabular reconstruction with use of morselized cryopreserved allogeneic cancellous bone graft and the Burch-Schneider ring can be highly successful in managing massive acetabular deficiencies in revision hip arthroplasty. PMID- 11407795 TI - Cementless acetabular reconstruction after acetabular fracture. AB - BACKGROUND: Total hip arthroplasty in patients with posttraumatic arthritis has produced results inferior to those in patients with nontraumatic arthritis. The use of cementless acetabular reconstruction, however, has not been extensively studied in this clinical context. Our purpose was to compare the intermediate term results of total hip arthroplasty with a cementless acetabular component in patients with posttraumatic arthritis with those of the same procedure in patients with nontraumatic arthritis. We also compared the results of arthroplasty in patients who had had prior operative treatment of their acetabular fracture with those in patients who had had prior closed treatment of their acetabular fracture. METHODS: Thirty total hip arthroplasties were performed with use of a cementless hemispheric, fiber-metal-mesh-coated acetabular component for the treatment of posttraumatic osteoarthritis after acetabular fracture. The median interval between the fracture and the arthroplasty was thirty-seven months (range, eight to 444 months). The average age at the time of the arthroplasty was fifty-one years (range, twenty-six to eighty-six years), and the average duration of follow-up was sixty-three months (range, twenty-four to 140 months). Fifteen patients had had prior open reduction and internal fixation of their acetabular fracture (open-reduction group), and fifteen patients had had closed treatment of the acetabular fracture (closed treatment group). The results of these thirty hip reconstructions were compared with the intermediate-term results of 204 consecutive primary total hip arthroplasties with cementless acetabular reconstruction in patients with nontraumatic arthritis. RESULTS: Operative time (p < 0.001), blood loss (p < 0.001), and perioperative transfusion requirements (p < 0.001) were greater in the patients with posttraumatic arthritis than they were in the patients with nontraumatic arthritis. Of the patients with posttraumatic arthritis, those who had had open reduction and internal fixation of their acetabular fracture had a significantly longer index procedure (p = 0.01), greater blood loss (p = 0.008), and a higher transfusion requirement (p = 0.049) than those in whom the fracture had been treated by closed methods. Eight of the fifteen patients with a previous open reduction and internal fixation required an elevated acetabular liner compared with one of the fifteen patients who had been treated by closed means (p = 0.005). Two of the fifteen patients with a previous open reduction and internal fixation required bone-grafting of acetabular defects compared with seven of the fifteen patients treated by closed means (p = 0.04). The thirty patients treated for posttraumatic arthritis had an average preoperative Harris hip score of 41 points, which increased to 88 points at the time of follow-up; there was no significant difference between the open-reduction and closed-treatment groups (p = 0.39). Twenty-seven patients (90%) had a good or excellent result. There were no dislocations or deep infections. The Kaplan-Meier ten-year survival rate, with revision or radiographic loosening as the end point, was 97%. These results were similar to those of the patients who underwent primary total hip arthroplasty for nontraumatic arthritis. CONCLUSIONS: The intermediate-term clinical results of total hip arthroplasty with cementless acetabular reconstruction for posttraumatic osteoarthritis after acetabular fracture were similar to those after the same procedure for nontraumatic arthritis, regardless of whether the acetabular fracture had been internally fixed initially. However, total hip arthroplasty after acetabular fracture was a longer procedure with greater blood loss, especially in patients with previous open reduction and internal fixation. Previous open reduction and internal fixation predisposed the hip to more intraoperative instability but less bone deficiency. PMID- 11407796 TI - Long-term results of total shoulder arthroplasty following bone-grafting of the glenoid. AB - BACKGROUND: The marked loss of glenoid bone volume or alteration of glenoid version can affect glenoid component fixation in patients undergoing total shoulder arthroplasty. The purpose of this study was to evaluate the long-term results associated with the use of bone-grafting for restoration of glenoid volume and version at the time of total shoulder arthroplasty. METHODS: Twenty one shoulders received an internally fixed, corticocancellous bone graft for the restoration of peripheral glenoid bone stock at the time of total shoulder arthroplasty between 1980 and 1989. Grafting was indicated when glenoid bone stock was insufficient to maintain adequate version or fixation of the prosthesis. Seventeen shoulders were available for follow-up; the average duration of follow-up for the thirteen shoulders that did not have prosthetic failure within the first two years was seventy months. Total shoulder arthroplasty was performed because of osteoarthritis in five shoulders, chronic anterior fracture-dislocation in five, capsulorrhaphy arthropathy in three, inflammatory arthritis in two, recurrent dislocation in one, and failure of a previous arthroplasty in one. All patients had some form of anterior or posterior instability preoperatively. There were five anterior and twelve posterior glenoid defects. Bone from the resected humeral head was used for grafting in fifteen shoulders, and bicortical iliac-crest bone was used in two. RESULTS: The average glenoid version after grafting was 4 degrees of retroversion, with an average correction of 33 degrees. The graft failed to maintain the original correction in three shoulders due to nonunion, dissolution, or shift. Five total shoulder replacements failed, necessitating glenoid revision at two to ninety-one months postoperatively. The failures were associated with recurrent massive cuff tears (one shoulder), persistent instability (two shoulders), improper component placement (one shoulder), and loss of graft fixation (one shoulder). There were no humeral component failures. According to the criteria of Neer et al., the functional result was rated as excellent in three shoulders, satisfactory in six, and unsatisfactory in eight. CONCLUSIONS: Despite the finding that eight shoulders had an unsatisfactory functional result at the time of longterm follow up, corticocancellous grafting of the glenoid successfully restored glenoid version and volume in fourteen of the seventeen shoulders in the present study. Patients with glenoid deficiency often have associated glenohumeral instability, which may affect the results of total shoulder arthroplasty. Bone-grafting of the glenoid is a technically demanding procedure that can restore bone stock in patients with structural defects. PMID- 11407797 TI - The effect of surgically implanted bullet fragments on the spinal cord in a rabbit model. AB - BACKGROUND: Whether or not to remove bullets or bullet fragments from the spinal column of a neurologically intact patient has been a subject of continual debate. The controversy is due in part to a lack of information about the long-term effects of bullet fragments on spinal cord tissue. Although many studies have demonstrated the toxic effects of metal fragments on brain tissue, to our knowledge no one has evaluated the effects of the metals contained in commercially available bullets on spinal cord tissue. METHODS: Copper, aluminum, and lead fragments from three commercially available bullet cartridges were implanted in intradural and extradural locations in seventeen New Zealand White rabbits. At an average of 9.8 months, the metal content of specimens of blood, cerebrospinal fluid, and liver were determined. The spinal cords were harvested and examined histologically. RESULTS: There was a significant increase in the copper level of blood from the rabbits with an implanted copper fragment compared with that of the control animals (p = 0.007). Concentrations of copper and lead were not elevated, compared with the control values, in the serum or liver. Histological examination of the spinal cords revealed major destruction of both the axons and the myelin of the dorsal column adjacent to the intradural copper fragments. Intradural fragments of lead caused similar destruction of myelin and axons in the dorsal column, but to a lesser degree. Minimal spinal cord or meningeal histological changes were noted around the aluminum intradural fragments, and no pathological changes were found near any fragments placed in an extradural location. CONCLUSIONS: The results of this study show that certain metals contained in commercially available bullets can cause varying degrees of neural destruction independent of the initial mechanical injury caused by implantation. Of the three metals tested, copper fragments consistently caused a substantial localized area of neural injury within the spinal cord. CLINICAL RELEVANCE: In our study, copper fragments caused local neural toxicity involving as much as 10% of the spinal cord area, suggesting that there may be a scientific basis for removal of copper fragments lodged in the spinal cord, even in the absence of a neurological deficit. PMID- 11407798 TI - Intrasynovial flexor tendon repair. An experimental study comparing low and high levels of in vivo force during rehabilitation in canines. AB - BACKGROUND: Rehabilitation methods that generate increased tendon force and motion have been advocated to improve results following intrasynovial flexor tendon repair. However, the effects of rehabilitation force and motion on tendon healing may be masked by the high stiffness produced by newer suture methods. Our objective was to determine whether the biomechanical properties of tendons repaired by one of two multistrand suture methods were sensitive to an increased level of applied rehabilitation force. METHODS: Two hundred and fourteen flexor digitorum profundus tendons from 107 adult dogs were transected and repaired. Dogs were assigned to one of four groups based on the rehabilitation method (low force [<5 N] or high force [17 N]) and the repair technique (four-strand or eight strand core suture) and were killed between five and forty-two days after the procedure. Repair-site structural properties were determined by tensile testing, and digital range of motion was assessed with use of a motion-analysis system. RESULTS: Tensile properties did not differ between the low and high-force rehabilitation groups, regardless of the repair technique (p > 0.05). In contrast, tensile properties were strongly affected by the repair technique, with tendons in the eight-strand group having an approximately 35% increase in ultimate force and rigidity compared with those in the four-strand group (p < 0.05). Ultimate force did not change significantly with time during the first twenty-one days (p > 0.05); there was no evidence of softening in either of the repair or rehabilitation groups. Force increased significantly from twenty-one to forty-two days, while rigidity increased throughout the forty-two-day period (p < 0.05). CONCLUSIONS: Increasing the level of force applied during postoperative rehabilitation from 5 to 17 N did not accelerate the time-dependent accrual of stiffness or strength. Suture technique was of primary importance in providing a stiff and strong repair throughout the early healing interval. CLINICAL RELEVANCE: Our findings suggest that there be a reexamination of the concept that increases in force produced by more vigorous mobilization protocols are beneficial to tendon-healing. While more vigorous rehabilitation may help to improve hand function, we found no evidence that it enhances tissue-healing or strength in the context of a modern suture repair. PMID- 11407799 TI - Prevention of venous thromboembolic disease following primary total knee arthroplasty. A randomized, multicenter, open-label, parallel-group comparison of enoxaparin and warfarin. AB - BACKGROUND: Patients treated with total knee arthroplasty are at high risk for the development of venous thromboembolism postoperatively. This study compared the efficacy and safety of two common thromboprophylactic agents, enoxaparin (a low-molecular-weight heparin) and warfarin. METHODS: Three hundred and forty-nine patients were included in a prospective, randomized, multicenter, open-label, parallel-group clinical trial. Treatment with enoxaparin (30 mg, administered subcutaneously twice daily) or warfarin (adjusted to an international normalized ratio of 2 to 3) was initiated during the immediate postoperative period, within eight hours after the surgery, and was continued for four to fourteen days. Venous thromboembolism was defined as deep-vein thrombosis documented by contrast venography, symptomatic deep-vein thrombosis documented by lower-extremity ultrasonography, or symptomatic pulmonary embolism confirmed by a positive lung scan or pulmonary angiography. RESULTS: In the all-treated-patients group, eighty (45%) of the 176 warfarin-treated patients had venous thromboembolism: fifty-nine (34%) had distal deep-vein thrombosis; twenty (11%), proximal deep-vein thrombosis; and one (0.6%), pulmonary embolism. Venous thromboembolism developed in significantly fewer (p = 0.0001) enoxaparin-treated patients (forty-four of 173; 25%): forty-one (24%) had distal deep-vein thrombosis, three (2%) had proximal deep-vein thrombosis, and none had pulmonary embolism. The enoxaparin treated patients also had a significantly lower prevalence of proximal deep-vein thrombosis (p = 0.002). The estimated odds for the development of venous thromboembolism were 2.52 times greater (95% confidence interval, 2.00 to 3.19) with warfarin than they were with enoxaparin. Major hemorrhage occurred in four warfarin-treated patients and nine enoxaparin-treated patients; with the numbers available, this difference was not significant (p = 0.17). Clinically important operative-site hemorrhage occurred in six (3%) of the warfarin-treated patients and twelve (7%) of the enoxaparin-treated patients (p = 0.15). CONCLUSIONS: A fixed 30-mg subcutaneous dose of enoxaparin, administered twice daily, with the first dose administered within eight hours after the completion of surgery, was significantly more effective than adjusted-dose warfarin in reducing the occurrence of asymptomatic venous thromboembolism, including proximal deep-vein thrombosis, in patients undergoing total knee arthroplasty. With the numbers available, there was no significant difference between groups with regard to the occurrence of major hemorrhagic complications; however, the rate of overall hemorrhagic complications was higher in the enoxaparin group. PMID- 11407800 TI - Effects of limb-length discrepancy on gait economy and lower-extremity muscle activity in older adults. AB - BACKGROUND: The amount of limb-length discrepancy necessary to adversely affect gait parameters in older adults is unknown, with information being largely anecdotal. This investigation was conducted to determine the effects of limb length discrepancy on gait economy and lower-extremity muscle activity in older adults. METHODS: Forty-four men and women ranging in age from fifty-five to eighty-six years with no evidence of limb-length discrepancy of >1 cm participated in the study. Subjects walked on a treadmill at a self-selected normal walking pace with artificial limb-length discrepancies of 0, 2, 3, and 4 cm applied in a randomly selected order. Indirect calorimetry was used to measure oxygen consumption and minute ventilation. Electromyography was used to measure muscle activity of the right and left quadriceps femoris, plantar flexors, gluteus maximus, and gluteus medius. Heart rate, the rating of perceived exertion, and frequency of gait compensation patterns were also measured. RESULTS: There was a significant increase in oxygen consumption and the rating of perceived exertion with 2, 3, and 4-cm artificial limb-length discrepancies; a significant increase in heart rate, minute ventilation, and quadriceps activity in the longer limb with 3 and 4-cm artificial limb-length discrepancies; and a significant increase in plantar flexor activity in the shorter limb with a 4-cm artificial limb-length discrepancy compared with the same parameters with no artificial limb-length discrepancy. CONCLUSIONS: Both oxygen consumption and the rating of perceived exertion were greater with a 2-cm artificial limb-length discrepancy than they were with no artificial limb-length discrepancy. There appears to be a breakpoint between 2 and 3 cm of artificial limb-length discrepancy in older adults with regard to the effects on most other physiological parameters. A 3-cm artificial limb-length discrepancy is likely to induce significant quadriceps fatigue in the longer limb. Elderly patients with substantial pulmonary, cardiac, or neuromuscular disease may have difficulty walking with a limb-length discrepancy as small as 2 cm. PMID- 11407801 TI - User's guide to the orthopaedic literature: how to use an article about a surgical therapy. PMID- 11407802 TI - The Academy on the edge: taking charge of our future. PMID- 11407803 TI - Orthopaedic information on the Internet. PMID- 11407804 TI - Arthroscopic repair versus open surgery for shoulder instability. PMID- 11407805 TI - Autologous blood and allogeneic transfusion. PMID- 11407806 TI - Attracting female candidates to the field of orthopaedic surgery. PMID- 11407807 TI - The Canadian orthopaedic residency experience. PMID- 11407808 TI - What's new in pediatric orthopaedics. PMID- 11407809 TI - Stereoselective total synthesis of chrysanthemol. AB - Chrysanthemol (1), a trans-eudesmane type sesquiterpene from Chrysanthemum indicum L., possesses certain anti-inflammatory activity. Its total synthesis was approached from two alternative routes and finally accomplished in ten steps from R-(+)-carvone via alpha-eudesmol (10) as the key intermediate. The overall yield is 2.4% and the spectral data of the synthetic target compound were identical with that of natural chrysanthemol (1). Seven intermediary compounds were tested for inhibitory effects on the carragenan-induced swelling of mouse paw but demonstrated no obvious activities. PMID- 11407810 TI - Sulfoglycolipid from the marine brown alga Sargassum hemiphylum. AB - One kinds of glycolipid (SBI) have been isolated from the marine brown alga Sargassum hemiphyllum (Turn.) Ag. The structures of SBI have been determined as the sodium salt of 1-0-acyl-3-0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol (acyl: tetradecanoyl, pentadecanoyl, 11-hexadecenoyl, hexadecanoyl, 10,13 octadecadienoyl, 9-octade cenoyl, 15-metylheptadecanoyl and 11-eicosenoyl 17: 1.5: 19: 153: 1: 19: 1: 2) on the basis of chemical and spectral evidence and GC MS analysis, respectively. Four constituents of the SBI were new compounds [the sodium salt of 1-0-(11"-hexadecenoyl)-3-0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol, the sodium salt of 1-0-(10",13"-octadecadienoyl)-3-0-(6'-sulfo-alpha-D quinovopyrannosyl) glycerol, and the sodium salt of 1-0-(15"-metylhexadecenoyl)-3 0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol, and the sodium salt of 1-0-(11" eicosenoyl)-3-0-(6'-sulfo-alpha-D-quinovopyrannosyl) glycerol]. All compounds were isolated from marine brown alga for the first time. PMID- 11407811 TI - Triterpenoid saponins from leaves and stems of Panax quinquefolium L. AB - In the chemical investigation on the saponin composition of leaves and stems of Panax quinquefolium L., two new minor dammarane saponins, quinquenoside L1 (1) and L2 (2) have been isolated. By means of physico-chemical evidences and spectral analysis their structures were established as 3-O-[beta-D-glucopyranosyl (1-2)-beta-D-glucopyranosyl]-20-O-beta-D-glucopyranosyl-dammara-23,25-diene 3beta, 12beta, 20(S)-triol (1) and 3-O-[beta-D-glucopyranosyl-(1-2)-beta-D glucopyranosyl]-20-O-beta-D-glucopyranosyl-(24Z)-dammar-24-ene-3beta, 12beta, 20(S), 26-tetraol (2). PMID- 11407812 TI - New norditerpenoid alkaloids from the roots of Aconitum geniculatum. AB - Four new norditerpenoid alkaloids, geniculatines A (1), B (4), C (7) and D (8), were isolated from the roots of Aconitum geniculatum Fletcher, and their structures were elucidated by spectral methods. PMID- 11407813 TI - Saikosaponin v-1 from roots of Bupleurum chinense DC. AB - Three triterpenoidal saponins, saikosaponin v-1(1), 6"-O-acetyl-saikosaponin b2 (2) and 6"-O-acetyl-saikosaponin d(3) were isolated from the roots of the title plant and the structures were identified on the basis of spectral analysis. Saikosaponin v-1 is a new compound, which was identified as 3beta,16alpha,23,28 tetrahydroxy-olean-11,13(18)-dien-30-oic acid-3-O-beta-D-glucopyranosyl-(1-->3) beta-D-fucopyranosyl-30-O-xylitol ester. PMID- 11407814 TI - New 2-(2-phenylethyl) chromones from Bothriochloa ischaemum. AB - Two new 2-(2-Phenylethyl) chromones were isolated from Bothriochloha ischaemum (Gramineae). They were characterized as 5-hydroxy-6-methoxy-2-[2-(2 hydroxyphenyl)-ethyl]-chromone(1) and 5-hydroxy-2-[2-(2-hydroxyphenyl)-ethyl] chromone(2) by means of spectroscopic analysis, especially 2D-NMR experiments. PMID- 11407815 TI - Cytochalasin D from Hypocrella bambusae. AB - Cytochalasin D which shows marked cytotoxic effects on multi-tumor cells was newly isolated at high content(5.28 mg/g, dry weight) from Fungus Hypocrella bambusae(B.et Br.) Sacc. Its structure was elucidated by spectroscopic methods. Two-dimensional NMR techniques were applied to make complete assignment for the 1H- and 13C-NMR chemical shifts of this compound. PMID- 11407816 TI - Platyphyllin A, a novel coumarone from the leaves of Betula platyphylla Suk. AB - A novel coumarone, platyphyllin A (1), was isolated from the leaves of Betula platyphylla Suk.. The structure elucidation was accomplished by the analysis of spectroscopic data. Compound 1 was identified as a new coumarone skeleton, which was first isolated from plants. PMID- 11407817 TI - C-27 and C-3 glucosylation of diosgenin by cell suspension cultures of Costus speciosus. AB - 3-O-[beta-D-glucopyranosyl-(1"--> 2')-beta-D-glucopyranosyl], 27-O-beta-D glucopyranosyl-(25R)-spirost-5-ene-3beta,27-diol was isolated from cell suspension cultures of Costus speciosus, following incubation with diosgenin, and its structure was elucidated using a combination of one- and two-dimensional 1H and 13C NMR spectral data, and positive and negative ion ESMS spectral data. PMID- 11407818 TI - Two novel phenolic triterpenes from Tripterygium wilfordii. AB - Two novel phenolic triterpenes were isolated from Tripterygium wilfordii Hook. f., their structures were identified to be 2,3-dihydroxy-1,3,5(10),8-tetra-ene 6alpha-(2'-hydroxyethyl)-24-nor-D:A-friedooleanane-29-oic acid 1, named triptotin F, and 2,3-dihydroxy-1,3,5(10),8-tetra-ene-6beta-(2'-hydroxyethyl)-24-nor-D:A friedooleanane-29-oic acid 2, named triptotin G on the basis of spectroscopic studies. PMID- 11407819 TI - Chestnutamide: a novel alkaloid from flowers of Castanea mollissima Blume. AB - A novel alkaloid, chestnutamide, was isolated from the flowers of Castanea mollissima Blume. The structure was determined on the basis of spectral analysis. PMID- 11407820 TI - Two new germacranolides from Magnolia grandiflora. AB - Two new sesquiterpenoids, 4,5-epoxy-13-methoxy-1(10)-germacren-12,6-olide and 4,5 epoxy-13-acetoxy-1(10)-germacren-12,6-olide, were isolated from the leaves of Magnolia grandiflora, together with six known compounds, 2alpha-hydroxy dihydroparthenolide, parthenolide, costunolide, syringaresinol, (+) medioresinol and 6,7-dimethoxycoumarin. The structures of the new compounds were elucidated on the basis of spectroscopic methods and X-ray diffraction. PMID- 11407821 TI - Electronic data collection at source for a joint replacement registry. PMID- 11407822 TI - Musculoskeletal images. Acetabular tuberculous osteomyelitis. PMID- 11407823 TI - Soft-tissue images. Retroperitoneal venous angioma. PMID- 11407824 TI - Musculoskeletal case 16. Fibrous dysplasia. PMID- 11407825 TI - Soft-tissue case 39. Air within the uterus and bladder. PMID- 11407826 TI - Advances in fluid resuscitation of hemorrhagic shock. AB - The optimal fluid for resuscitation in hemorrhagic shock would combine the volume expansion and oxygen-carrying capacity of blood without the need for cross matching or the risk of disease transmission. Although the ideal fluid has yet to be discovered, current options are discussed in this review, including crystalloids, colloids, blood and blood substitutes. The future role of blood substitutes is not yet defined, but the potential advantages in trauma or elective surgery may prove to be enormous. PMID- 11407827 TI - Prognostic markers in resectable non-small cell lung cancer: a multivariate analysis. AB - OBJECTIVE: To identify the prognostic significance of certain clinical, cellular and immunologic markers in resectable non-small cell lung cancer (NSCLC). DESIGN: A cohort of patients with resectable NSCLC was prospectively followed up for 8 years (100% follow-up). SETTING: A university hospital in a large Canadian city. PATIENTS: One hundred and thirteen consecutive patients who underwent surgical resection of primary NSCLC. MAIN OUTCOME MEASURES: Presence of peritumoral B lymphocytes (identified with antibody to CD20) and T lymphocytes (antibody to CD43), along with tumour markers (carcinoembryonic antigen [CEA], keratin, cytokeratin, S-100 protein, vimentin, chromogranin) and other factors such as age, sex, cell type, American Joint Committee on Cancer (AJCC) stage, histologic grade, DNA ploidy and S-phase fraction were correlated with survival. RESULTS: The mean age of patients in the study was 66.0 years; 60% were male. Histologic types of the tumours were: adenocarcinoma 57 (50.4%), squamous cell 47 (41.6%), adenosquamous 6 (5.3%) and large cell 3 (2.6%). AJCC stages were: I 66 (58.4%), II 20 (17.7%) and III 27 (23.9%). Histologic grades were: I (well differentiated) 31 (27.4%), II 50 (44.2%), III 29 (25.7%) and IV 3 (2.6%). Survival was 85% at 1 year (95% confidence interval [CI] 76%-90%), 44% at 5 years (95% CI 34%-53%) and 34% at 10 years (95% CI 22%-46%). Multivariate analyses using the Cox proportional hazards model for survival confirmed AJCC stage (p < 0.001) in all histologic subtypes to be the strongest factor of independent prognostic significance. It also revealed the presence of CD20-stained B lymphocytes (p = 0.04) in the peritumoral region of all tumours to be a positive prognostic factor. This relation was especially strong for nonsquamous cell carcinomas (p < 0.001). For squamous cell carcinomas, the immunohistochemical presence of CEA was of marginally negative prognostic value (p = 0.04). DNA ploidy and a high S-phase fraction showed no evidence of prognostic value for stage I tumours, but for stages II and III tumours there was strong evidence of prognostic value (p < 0.001 jointly). The evidence for DNA ploidy was especially strong in stages II and III squamous cell tumours (p = 0.008), and for a high S-phase fraction was strongest in stages II and III nonsquamous cell tumours (p = 0.002). CONCLUSIONS: AJCC stage remains the most important prognostic indicator from a variety of clinical variables and tumour markers in postoperative patients with resectable NSCLC. For nonsquamous cell lung carcinomas, the presence of peritumoral B lymphocytes was strongly associated with improved survival, suggesting an important role for humoral mediated immunity. PMID- 11407828 TI - Isolated limb infusion for melanoma: a simple alternative to isolated limb perfusion. AB - OBJECTIVE: To describe initial experience with the new technique of isolated limb infusion (ILI) for in-transit melanoma. DESIGN: A prospective case series. SETTING: The major tertiary care oncology centre for the state of Victoria, Australia. PATIENTS: Nine patients having for extensive in-transit limb melanoma INTERVENTIONS: All patients received ILI (13 treatments). OUTCOME MEASURES: Patient survival, response to treatment and complications of treatment. RESULTS: There were no perioperative deaths and morbidity was limited to deep venous thrombosis and pulmonary embolism in 1 patient. Control of the in-transit metastases was achieved to some degree in all patients and was complete in 4. CONCLUSIONS: ILI is an alternative treatment modality for patients suffering from multiple, advanced in-transit melanoma metastases. It provides effective palliation with limited morbidity and offers a safe, quick, inexpensive alternative to isolated limb perfusion with comparable results. PMID- 11407829 TI - Analysis of the efficacy of pediatric day surgery. AB - OBJECTIVES: To assess the efficacy and safety of a pediatric day surgery program and its benefits to the child and family. DESIGN: A review and analysis of prospectively gathered data. SETTING: The Children's Hospital of Eastern Ontario (CHEO), a university-affiliated pediatric centre with a referral base of 2.5 million people. PATIENTS: All children undergoing day surgery at CHEO during the 5 years between 1992 and 1997. INTERVENTIONS: All surgical procedures performed on a day surgery basis. MAIN OUTCOME MEASURES: Procedures performed, complications and unexpected admissions. RESULTS: An average of 4,899 children per year underwent surgical procedures in the ambulatory day surgery program. The 4 commonest procedures performed were myringotomy, tonsillectomy and adenoidectomy, dental procedures, and inguinal hernia repair. The majority of children were between 2 and 7 years of age. Complications were few, averaging 1.6% per year, with postoperative bleeding, primarily secondary to tonsillectomy and adenoidectomy, being the most common. None resulted in permanent disability and there were no deaths. CONCLUSIONS: Pediatric day surgery is a safe and cost effective program that benefits the child, the family and the pediatric surgical program. PMID- 11407830 TI - The Southwestern Ontario Joint Replacement Pilot Project: electronic point-of care data collection. Southwestern Ontario Study Group. AB - OBJECTIVE: To pilot a provincial joint replacement registry using electronic point-of-care data collection. DESIGN: Data collection study. SETTING: Southwestern Ontario, which has a population base of 3.5 million people. PARTICIPANTS: Eighteen orthopedic surgeons. METHOD: Information on total hip and knee replacements was obtained by the orthopedic surgeons over a 6-month period. Information was obtained in paper form and electronically on hand-held computers. MAIN OUTCOME MEASURES: Patient demographics, waiting times from referral to operation, patient satisfaction and relevance and value of electronic records compared with paper records. MAIN RESULTS: Data were collected on 815 total hip and knee arthroplasties. A slightly greater number of hips required revision than knees. The majority of patients were in the 60 to 90-year age range. With respect to the waiting time from referral to operation 10% of patients waited less than 5 weeks, 50% waited less than 30 weeks, and 90% waited less than 59 weeks. There was a high level of patient satisfaction with the operation and with hospital care received. Most surgeons found that the gathering and use of data electronically was relevant and easy. The electronic data were more timely, accurate and complete than paper records. CONCLUSION: Electronic point-of-care data collection is appropriate, particularly in high-volume, high-cost surgical interventions such as total joint replacements. PMID- 11407831 TI - The management of liver hydatid cysts by percutaneous drainage. AB - OBJECTIVE: To investigate the effect of percutaneous drainage on liver hydatid cysts. DESIGN: A retrospective case study. SETTING: Department of Surgery, Selcuk University, Konya, Turkey. PATIENTS: Forty-five patients with 83 liver hydatid cysts (types I and II according to the classification of Gharbi and colleagues) followed up for a mean of 30 months (range from 14 to 36 months). INTERVENTION: The cysts were drained percutaneously with ultrasonographic guidance and then irrigated with 0.05% silver nitrate solution through a fine needle. Albendazole was administered 48 hours before percutaneous drainage and for 2 months after the procedure to prevent the implantation of spilled scolices. MAIN OUTCOME MEASURES: Complications of the procedure, decrease in size of the cyst cavity, recurrence and dissemination of the cysts. RESULTS: All the cysts were treated successfully by percutaneous drainage. Anaphylactic shock developed in 1 (2.2 %) patient, and mild allergic reactions were observed in 2 (4.4 %) patients during the interventional procedure. Follow-up ultrasonography and CT demonstrated a statistically significant (p < 0.01) decrease in the mean cyst size. Recurrence and dissemination were not observed during the follow-up period. CONCLUSION: Percutaneous fine-needle aspiration and drainage is effective for managing cystic liver hydatid disease in selected cases. PMID- 11407832 TI - Practising physician's knowledge and patterns of practice regarding the asplenic state: the need for improved education and a practical checklist. AB - OBJECTIVE: To examine physicians' knowledge and actions regarding the asplenic state and to develop a practical checklist to aid in the systematic education and management of asplenic patients. DESIGN: A prospective cohort survey utilizing an experienced nurse practitioner and a survey questionnaire with on-site interviews. SETTING: The Okanagan Valley, British Columbia. SUBJECTS: A cohort of 122 physicians serving a population base of 350,000. MAIN OUTCOME MEASURES: Beliefs and practices relating to vaccination and precautions necessary for adult and pediatric splenectomized patients. PRINCIPAL RESULTS: The majority of physicians appeared to be knowledgeable about potential conditions affecting splenic function, except in the case of severe liver disease with portal hypertension and collagen vascular disease. There appeared to be good understanding on the part of most physicians of the risks associated with various infectious diseases and the asplenic state, except in the case of Capnocytophaga canimorsus infection linked to dog bites and the increased susceptibility of asplenic patients to intraerythrocytic parasites. Although a majority of physicians were cognizant of the need for pneumococcal vaccination and other immunizations in adults, there was marked uncertainty in relation to the need and the appropriate time interval for revaccination. In the case of children there appeared to be uncertainty regarding the role of antibiotic prophylaxis. There were discrepancies between physicians' expressed attitudes and the actions actually taken for asplenic patients in individual practices. CONCLUSIONS: Further education is required concerning the management of asplenic patients. The systematic use of a practical checklist may facilitate this process. PMID- 11407833 TI - Pulmonary metastases from colorectal cancer: 25 years of experience. AB - OBJECTIVE: To examine the experience of pulmonary resections for colorectal metastases at the McGill University Health Centre. DESIGN: A chart review. PATIENTS: Forty-nine patients treated surgically between 1975 and 1998 for pulmonary metastases from colorectal cancer. INTERVENTION: Thoracotomy with pulmonary resection. OUTCOME MEASURES: Survival of patients with various preoperative and post operative clinical variables. RESULTS: The perioperative death rate was 4%. Overall 5- and 10-year survival rates were 55% and 40% respectively. The mean interval between the initial colonic resection and resection of pulmonary metastases (discase-free interval) was 36 months. The 7 patients who also under went resection of extrapulmonary metastases had a 5-year survival rate of 52%. Significant preoperative variables that carried a poor prognosis included the following: more than one pulmonary lesion, a disease-free interval of less than 2 years, and moderately or poorly differentiated colorectal cancer. The 16 patients who received chemotherapy after their thoracotomy had a 5 year survival rate of 51% compared with 54% for the 33 patients who did not receive chemotherapy. Recurrent resections of pulmonary lesions did not reduce survival. CONCLUSIONS: Pulmonary resection for metastatic colorectal cancer is both effective and safe. Resectable extrapulmonary metastases and pulmonary recurrence should not preclude lung resection. Postoperative chemotherapy has no survival benefit. Preoperative variables should guide the clinician when considering surgical intervention. PMID- 11407834 TI - Multicentric giant cell tumour involving the patella. PMID- 11407835 TI - Extrapelvic endometriosis associated with occult groin hernias. PMID- 11407836 TI - Coexisting mammary tuberculosis and malignant disease. PMID- 11407837 TI - Education versus service. PMID- 11407838 TI - Unique presentation of a bronchial foreign body in an asymptomatic child. AB - A child who aspirates a foreign body may present in many ways. We present the case of an 8-year-old boy whose initial presentation consisted only of his preoccupation with his lost toy and his gesturing toward his oral cavity. There was no witnessed coughing or choking episode, and there were no initial physical examination findings of note, and no abnormalities on appropriate radiographic studies. The patient was transferred to our institution for further evaluation and management. Our careful examination revealed a slight wheeze at the left upper lobe. Endoscopy was immediately performed, and bronchoscopy revealed a yellow, tubular, plastic foreign body in the left main bronchus through which the patient was breathing freely. This was removed without incident by means of optical graspers. The physical characteristics of this foreign body, including its small size, relatively inert material, and large lumen, allowed this patient to present relatively asymptomatically. This case demonstrates the need for a high index of suspicion in the evaluation and management of foreign bodies in the aerodigestive tract. PMID- 11407839 TI - Open Zenker's diverticulectomy using stapling techniques. AB - This is a prospective review of patients who underwent Zenker's diverticulectomy, closure of the pharyngotomy with nonabsorbable staples, and cricopharyngeal myotomy through an open neck approach over a period of 2 years. The study group consisted of 9 men with a mean age of 67 years. The diagnosis of Zenker's diverticulum was based on a barium swallow esophagram. The mean operative time was 1.5 hours, with a mean estimated blood loss of 25 mL. Barium swallow esophagrams performed on the first postoperative day showed no pharyngeal fistulas. Oral feeding was started after the esophagram. One patient was discharged on postoperative day 1, 1 was discharged on postoperative day 3, and the remaining 7 were discharged on postoperative day 2. All patients were able to take oral feeding by postoperative day 1. There were no operative complications in this series. We conclude that stapling with nonabsorbable staples is a relatively safe and effective method for closing pharyngeal defects after Zenker's diverticulectomy. The technique leads to shortening of the operative time and hospital stay, and allows early resumption of oral feeding. PMID- 11407840 TI - Mapping of brain stem neuronal circuitry active during swallowing. AB - A poorly understood neural circuit in the brain stem controls swallowing. This experiment studied the swallowing circuit in the rat brain stem by means of fos immunocytochemistry. The fos protein is a marker of activated neurons, and under experimental conditions, repetition of a behavior causes the fos protein to be produced in the neurons involved in that behavior. The fos technique has been successfully used to delineate neural circuits involved in reflex glottic closure, cough, and vocalization; however, the technique has not been used to map the swallowing circuit. Nine rats were used in this study. Swallows were evoked in anesthetized rats for 1 hour, then, after a 4-hour delay to allow maximum fos production, the rats were painlessly sacrificed by perfusion. The brain stems were removed and sectioned in the frontal plane, and every fourth section was immunoreacted for fos protein. All sections were examined by light microscopy, and cells positive for fos were marked on drawings of brain stem structures for different levels throughout the brain stem. Control animals underwent sham experiments. After subtraction of the areas of fos labeling seen in controls, all experimental rats showed fos-labeled neurons in very discrete and localized areas, including practically all regions implicated by prior neurophysiology studies of swallowing. The distribution of labeled neurons was more dispersed through the brain stem than current theories of swallowing would suggest. Specifically, recent studies of swallowing control have focused on the nucleus of the solitary tract (NST) and the region surrounding the nucleus ambiguus (periambigual area) just rostral to the obex. These areas contained fos-labeled neurons, but unexpectedly, heavy labeling was found in the same areas caudal to the obex. Areas containing the heaviest labeling were specific subnuclei of the NST and surrounding reticular formation; the periambigual area; and the intermediate reticular zone in the pons and caudal medulla. Interestingly, none of these anatomic structures had uniform fos labeling; this finding suggests that the unlabeled areas are involved in other oromotor behaviors, or that the specific protocol did not activate the full population of swallowing-related neurons. A notable finding of this study is a candidate for the central pattern generator (CPG) of swallowing. Careful lesioning studies in cats strongly suggest that a region in the rostral-medial medulla contains the CPG for swallowing, although the exact location of the CPG was never pinpointed. In the homologous region of the rat brain stem, fos labeling was only found in a small group of neurons within the gigantocellular reticular formation that may be a candidate for the CPG. In summary, correlation with prior physiology experiments suggests that this experiment appears to have delineated many, if not all, of the components of the swallowing circuit for the first time in any mammal. In addition, other areas were found that might also be swallowing-related. One notable example is a small group of fos-labeled cells that may be the CPG for swallowing. Further studies are required to clarify the specific roles of the fos labeled neurons seen in this study. PMID- 11407841 TI - Hemicricoidectomy for voice rehabilitation following hemilaryngectomy with ipsilateral arytenoid removal. AB - The purpose of this article is to describe an approach to reconstruction of the larynx after vertical partial laryngectomy with removal of the ipsilateral arytenoid cartilage. This method addresses the problem of postoperative posterior glottal incompetence (aphonia with or without aspiration). The technique involves resection of the ipsilateral half of the cricoid cartilage, use of an inferiorly based strap muscle flap for vocal fold reconstruction, and placement of a customized stent. This technique may be used at the time of the primary cancer extirpation or as a secondary rehabilitative procedure. Generally, patients who have undergone this procedure have had minimal postoperative breathiness with good phonatory and airway function. We recommend this reconstructive technique for patients with large posterior defects following hemilaryngectomy. PMID- 11407842 TI - Anterior window laryngoplasty: a new anatomic approach for posterior glottic and subglottic stensosis. AB - This study explores the feasibility of an anterior thyroid window as a new approach for posterior glottic stenosis. An anterior window was made on the thyroid cartilage of human cadaver larynges, with its superior edge below the level of the vocal cords and an inferior pedicle composed of the inferior border of the thyroid cartilage attached to the cricothyroid membrane, which was left intact. By obtaining direct exposure of the posterior glottis and cricoarytenoid joints, mucosal graft suturing and exploration of the joints were easily achieved with an operating microscope. This study demonstrates the anatomic feasibility of an anterior window laryngoplasty as an alternative approach to treating posterior glottic stenosis while preserving the cartilage framework and avoiding the anterior commissure. This technique may improve endolaryngeal manipulation by providing a closer and more direct exposure than do endoscopic techniques, and by having fewer possible complications than traditional open techniques. PMID- 11407843 TI - Intraoperative pediatric laryngeal electromyography: experience and caveats with monopolar electrodes. AB - We report our experience with intraoperative laryngeal electromyography (L-EMG) using direct laryngoscopy and placement of monopolar electrodes under general anesthesia in the evaluation and management of laryngeal dysfunction in pediatric patients. In this series of case studies, we present clinical data on 30 pediatric patients with known or suspected anatomic or neurologic laryngotracheal disorders evaluated with placement of shielded monopolar electrodes into the thyroarytenoid muscles during direct laryngoscopy under general anesthesia. Diagnoses included congenital vocal fold paralysis (VFP), laryngotracheal stenosis, cerebral palsy, laryngeal tumors, traumatic vocal fold dysfunction, and postsurgical VFP. The impact of L-EMG on patient management was assessed. We found that L-EMG objectively supported clinical findings, but provided new objective data relevant toward management recommendations in only a few selected pediatric patients with new-onset vocal fold paralysis or paresis or infiltrative laryngeal tumors, and in selected postsurgical cases involving decannulation decisions. The prognostic utility of L-EMG in newborns with congenital VFP has not been established. A normal L-EMG recording indicates an intact neuromuscular axis, but does not guarantee vocal fold mobility or guarantee muscle function in a partially denervated or deconditioned muscle. The potential for false-negative recordings is the major limitation of this technique. PMID- 11407844 TI - Laryngeal reinnervation with the hypoglossal nerve. I. Physiology, histochemistry, electromyography, and retrograde labeling in a canine model. AB - This study was performed to determine whether the hypoglossal nerve (cranial nerve XI [XII]) would serve as a useful donor for laryngeal reinnervation by anastomosis to the recurrent laryngeal nerve (RLN). Twenty hemilarynges in 10 dogs were studied prospectively after XII-RLN anastomosis (group A; n = 5), split XII-RLN anastomosis (group B; n = 3), XII-RLN anastomosis with a 2-cm interposition graft (group C; n = 2), no treatment (group D; n = 5), RLN section (group E; n = 2), or ansa cervicalis-RLN anastomosis (group F; n = 3). Spontaneous activity was observed monthly by infraglottic examination through permanent tracheostomies and was recorded by electromyography. Laryngeal adductory pressure and induced phonation were obtained by stimulating the RLN while passing a pressure transducer balloon or humidified air through the glottis. At sacrifice, the laryngeal muscles were stained for adenosine triphosphatase to determine the ratio of type I to type II fibers. Retrograde labeling of the brain stem was performed with horseradish peroxidase. Infraglottic examination at 6 months showed a full range of adductory motion in groups A and B during the swallow reflex, comparable with that in group D. Groups C and F showed good bulk and tone, but little spontaneous motion. Group E remained paralyzed. Stimulation of the transferred nerves caused more activity in groups A and B than in the other groups; groups C and F partially adducted at high levels. The laryngeal adductory pressure responses of groups A and B were similar to those of group D. The XII-reinnervated larynges were capable of producing normal induced phonation. Retrograde labeling of the RLN showed that the reinnervating axons originated only in the hypoglossal nucleus. Electromyography of the reinnervated adductor muscles confirmed spontaneous activity in the dogs (awake). Histochemical analysis confirmed slow-to-fast transformation of both the posterior and lateral cricoarytenoid muscles, indicating that significant reinnervation occurred. We conclude that the hypoglossal nerve functions well as a donor for adductory reinnervation of the larynx. PMID- 11407845 TI - Combined buccal mucosa island and sternohyoid flaps: a new technique of hemilaryngeal reconstruction studied in a canine model. AB - No single method of reconstruction has proven ideal for all patients with defects following vertical hemilaryngectomy. In this report, we detail a new technique for hemilaryngeal reconstruction involving the use of a pedicled buccal mucosa island flap supplied by the facial artery and vein. The buccal flap was used to resurface a transversely oriented sternohyoid myofascial flap. The reconstructive outcome was analyzed in 4 animals, 3 of which survived the early postoperative period. Videoendoscopy and stroboscopy were performed to analyze the laryngeal configuration and vibration. Each subject was decannulated and had a competent airway free of aspiration. After sacrifice of the animals, whole organ axial sections were made at multiple levels. Endoscopic and histologic findings documented that this technique produced an appropriate neocord position. Laryngeal stroboscopy in each animal showed bilateral mucosal traveling waves, with entrainment of the reconstructed neocord mucosa and native vocal cord mucosa. We conclude that the layered reconstructive technique described, compared to traditional methods of reconstruction, more closely replicates the structure of the excised tissue in hemilaryngeal reconstruction, potentially resulting in an improved voice outcome. PMID- 11407846 TI - Aerodynamic profiles of a hemilarynx with a vocal tract. AB - The pressure-flow relationship was examined in excised canine and human larynges with and without a vocal tract. Canine and human larynges were prepared and cut in the midsagittal plane from the top to about 10 mm below the vocal folds. The right half was removed and replaced with an acrylic plate. The vocal tract was simulated initially with a 15-cm plastic tube and later with a vocal tract model with an area function resembling "ah." Simultaneous recordings were made of the glottal pressure, mean subglottal pressure, and average airflow at various levels of adduction. Preliminary data indicated that the pressure-flow relationships were similar to those of a full larynx and were almost linear. The addition of the vocal tract increased the glottal resistance by moving these pressure-flow lines to the lower-flow and higher-pressure region. The human larynx appears to phonate more easily than the canine larynx on the laboratory bench and has lower phonation threshold pressures. PMID- 11407847 TI - Effects of length and depth of vibration of the vocal folds on the relationship between transglottal pressure and fundamental frequency of phonation in canine larynges. AB - The relationship between fundamental frequency of phonation (F0) and the change in F0 per unit change of transglottal pressure (dF/dP) was investigated in 3 excised canine larynges. The effects of the length and depth of vocal fold vibration on the relationship between dF/dP and F0 were evaluated. A positive correlation between dF/dP and F0 was observed when an increase in F0 was accompanied by a decrease in the depth of vibration while the length of vibration reached a plateau. Although a negative correlation between dF/dP and F0 was observed in only 1 case, this relationship seemed relevant to the length of vibration. These results help to clarify the adjustments of length and depth of vibration involved in regulating F0. PMID- 11407848 TI - Isolated vestibular areflexia after blunt head trauma. AB - The sudden unilateral loss of vestibular function is a frequent cause of vertigo. This condition is called vestibular neuronitis or vestibular neuritis. Its cause remains unknown, but many authors consider it to be a sequel of vestibular viral infection. We report the history and clinical findings of 5 patients in whom a unilateral vestibular loss occurred after head trauma. None of these patients complained of hearing loss. In all cases, the vertigo gradually subsided over days or weeks. The follow-up showed the partial recovery of vestibular function in 2 cases, while vestibular areflexia persisted in 3. The clinical course and findings were similar in every respect to those in patients with classic idiopathic vestibular neuronitis. PMID- 11407849 TI - Cochlear microphonic potentials: a new recording technique. AB - A new instrumentation and a particular method for detecting and recording cochlear microphonic potentials (CMPs) are described here. The CMPs were recorded in rats by means of pure tones (4,000, 2,000, 1,000, 500, and 250 Hz) and intraepidermic electrodes; the electrocochleography technique was avoided. An experimental design that included the use of a glutamatergic agonist (kainic acid [KA]) and an aminoglycoside antibiotic (kanamycin [KANA]) was carried out to demonstrate the origin of the recorded potential. Morphological studies showed that KA selectively eliminated the afferent type I dendrites of the spiral ganglion, while the administration of KANA resulted in the absence of outer hair cells. When CMPs were recorded after KA administration, no alterations were detected. In contrast, KANA administration resulted in the absence of any selective electrophysiological activity corresponding to CMPs. All these results were compared with the recording of the compound action potential of the eighth nerve obtained by electrocochleography. These findings and the great specificity of the reproduction of the sound stimulus confirm that the CMPs can be recorded by the new equipment. PMID- 11407850 TI - Role of interleukin-1beta in a murine model of otitis media with effusion. AB - To clarify the role of interleukin-1beta (IL-1beta) in the pathogenesis of otitis media with effusion (OME), we developed and investigated a murine model of this disease. Specific pathogen-free male BALB/c mice received intratympanic injections of 20 microg of endotoxin derived from nontypeable Haemophilus influenzae. Three days after injection, middle ear effusions were observed through the eardrum. Similar pathological changes were observed after inoculation with 100 ng of recombinant IL-1beta. Anti-IL-1 receptor antibodies inhibited the pathological changes induced by the endotoxin. In situ hybridization showed expression of IL-1beta messenger RNA in the epithelium of the middle ear mucosa. These results suggest that IL-1beta might be associated with endotoxin-induced inflammation in the middle ear and might play an important role in the induction of otitis media with effusion. PMID- 11407851 TI - Bell's palsy: electrodiagnostics are not indicative of cerebrospinal fluid abnormalities. AB - Electrodiagnostic testing (electromyography, electroneuronography, and blink reflex) and cerebrospinal fluid (CSF) examination (cell count, immunoglobulins, and antigen-specific intrathecal immunoglobulin G synthesis against herpes simplex virus, varicella zoster virus, cytomegalovirus, and Borrelia burgdorferi sensu latu) were performed in 56 patients with Bell's palsy. The CSF was normal in 45 patients and abnormal in 11 patients. Acute borreliosis was the most common specific pathological CSF finding (4 of 11). Electromyography revealed abolished volitional activity in 22% of patients with normal CSF and in 36% with pathological CSF. Electroneuronographic tests with an amplitude decrease of more than 90% on the affected side or abolished responses were found in 20% of patients with normal CSF and in 18% with pathological CSF. Abolished orbicularis oculi reflexes were seen in 67% of patients with normal CSF and in 82% with pathological CSF Concerning electrodiagnostic testing, no statistically significant difference between patients with normal and abnormal CSF was found, so we conclude that electrodiagnostic testing has no indicative value for abnormal CSF in Bell's palsy. PMID- 11407852 TI - Topical ascorbic acid reduces myringosclerosis in perforated tympanic membranes. A study in the rat. AB - Myringosclerosis, a common finding after myringotomy, has been recently associated with an increased production of oxygen free radicals. Ascorbic acid's proposed actions include collagen synthesis, antioxidation, and free radical scavenging. The effects of topical ascorbic acid on healing tympanic membranes were studied. Particular attention was given to detecting the presence of myringosclerosis. Twelve Sprague-Dawley rats were bilaterally myringotomized. Their ears were randomized into group A, which received topical ascorbic acid in Gelfoam, group B, which received topical saline solution in Gelfoam, and group C, which received no treatment. The tympanic membranes were harvested on day 13, after routine otomicroscopy. Under light microscopy, the connective tissue layer of the untouched side of the pars tensa was distinctly thicker in group A than in group B or group C. At this level, the extent of sclerotic lesions was significantly less in the ascorbic acid-treated group. It is inferred that topical ascorbic acid reduces the occurrence of myringosclerosis following tympanic membrane perforations in the rat. PMID- 11407853 TI - Intracranial complications of ethmoiditis evidenced by magnetic resonance imaging. PMID- 11407854 TI - Experience with StatView 5.0. PMID- 11407855 TI - X-ray microanalysis of etoposide-induced apoptosis in the PC-3 prostatic cancer cell line. AB - Apoptosis comprises a critical intracellular defense mechanism against tumourigenic growth. We have been interested in the relationship between morphological changes and intracellular concentration of several cations after etoposide-induced apoptosis in androgen-independent prostate cancer cells. SEM and X-ray microanalysis were performed on freeze-dried PC3 cells after etoposide treatment, and correlated with the morphological features observed after examination by light and fluorescence microscopy. Cell viability assays were also performed. A significant decrease in intracellular Cl(-) and K(+)and a progressive increase in Mg(2+) and Na(+) were observed, with parallel changes in cellular volume as cells passed through three morphological stages of apoptosis. The use of EPXRMA made it possible to evaluate alterations in element composition in prostate cancer cell apoptosis and may be a helpful tool for further studies on apoptosis in prostate cancer. PMID- 11407856 TI - Cell death in Tetrahymena thermophila: new observations on culture conditions. AB - We previously suggested that the cell fate of the protozoan ciliate, Tetrahymena thermophila, effectively relates to a quorum-sensing mechanism where cell released factors support cell survival and proliferation. The cells have to be present above a critical initial density in a chemically defined nutrient medium in order to release a sufficient level of these factors to allow a new colony to flourish. At a relatively high rate of metabolism and/or macromolecular synthesis and below this critical density, cells began to die abruptly within 30 min of inoculation, and this death took the form of an explosive disintegration lasting less than 50 milliseconds. The cells died at any location in the culture, and the frequency of cell death was always lower in well-filled vials than those with medium/air interface. Cell death was inhibited by the addition of Actinomycin D or through modifications of the culture conditions either by reducing the oxygen tension or by decreasing the temperature of the growth medium. In addition, plastic caps in well-filled vials release substances, which promote cell survival. The fate of low-density cultures is related to certain 'physical' conditions, in addition to the availability of oxygen within closed culture systems. PMID- 11407857 TI - Al(3+) and Zn(2+)-induced structural changes and localization in a fully-hydrated live eukaryotic cellular model. AB - This study combined techniques that did not require preparation protocols that were potentially harmful to the cell, making it possible to investigate cells at, or close to, their natural physiological state. We used the freshwater protozoon Chilomonas paramecium as a eukaryotic cellular model to locate sites of Al(3+) or Zn(2+) accumulation and quantify the associated structural changes. Cells were fully hydrated throughout the study, which used a combination of differential interference contrast light microscopy, confocal laser scanning microscopy and transmission X-ray microscopy. The latter technique allowed high resolution (50 nm) and high contrast imaging of live cells in solution. For confocal laser scanning microscopy the relatively new fluorochrome Newport Green was used. This made fluorescent complexes with intracellular Al(3+) and Zn(2+), allowing localisation of metal-containing granules and vesicles. After long term exposure a previously unreported annular-shaped site of metal accumulation was found, signifying a vesicle with metal accumulated in the periphery only. After exposure to Al(3+) and Zn(2+), the cell pellicle was thinner and the majority of rounded up cells had a concentric layering of organelles. By combining a variety of techniques it was possible to gain high resolution structural and chemical information on cells minimally exposed to potentially artefact-inducing procedures. PMID- 11407858 TI - Restoration of the rat urothelium after cyclophosphamide treatment. AB - Processes leading to the recovery of a normal three-layered urothelium from a hyperplastic urothelium induced by cyclophosphamide (CP) treatment in rats have been investigated. A single intraperitoneal (ip) dose of CP caused extensive loss of cells from urothelium, but the remaining cells started to express epidermal growth factor receptor (EGFR) in their plasma membranes. On day 2 after CP injection, proliferating cell nuclear antigen (PCNA) immunohistochemistry showed a rapid increase in positively stained nuclei, from which a hyperplastic urothelium developed, composed of undifferentiated cells expressing EGFR over the entire plasma membrane. Subsequently, EGFR gradually disappeared from the apical plasma membrane but remained in the basolateral membranes. After day 6, PCNA positive nuclei in all cell layers decreased, except in basal cells. Apoptotic cells were detectable by the TUNEL assay at day 2, and increased in number in all layers of the hyperplastic urothelium until day 10, returning to the control levels by day 14. Electron microscopic evidence showed that apoptotic cells were either pinched off into the bladder lumen or phagocytosed by the neighbouring urothelial cells. Thus, the urothelium responds to the damage by intense proliferation for a week, resulting in an undifferentiated hyperplastic state. Differentiation of superficial cells then begins and damaged cells are gradually removed by apoptosis until the three-layered urothelium is fully restored by two weeks following CP treatment. PMID- 11407859 TI - Not all muscles meet the same fate when they die. AB - Two general patterns of cell death are usually described in animals: necrosis and apoptosis. The former is a passive process that displays cellular swelling and lysis, while the latter involves cellular shrinkage and gene-mediated, ATP dependent processes. Independent of the proximal cause of cell death, cell corpses are almost always removed by phagocytic cells. This is far from universal for all cells however, since phagocytic cells have not been noted during the programmed death of some skeletal muscles in insects. To further explore this, we used a variety of anatomical methods to examine the death of the intersegmental muscles (ISMs) of the moth Manduca sexta. The ISMs are giant cells that die during the 30 h following adult emergence. At no stage examined were hemocytes or other cells associated with the sarcolemma. The failure to detect macrophages was not due to technical limitations since immunohistochemical and functional studies demonstrate their presence in the hemolymph. The absence of phagocytosis to remove ISM corpses suggests that all of the biochemical machinery required for cellular destruction is resident within the ISMs themselves. This is consistent with analysis suggesting that Manduca does not possess sufficient numbers of macrophages to consume the ISMs. Given that insects do not have adaptive immunity, the ability to use a completely cell autonomous process may be a developmental option that cannot be exploited in vertebrates. PMID- 11407860 TI - Reduced expression of the BRCA1 gene and increased chromosomal instability in MCF 7 cell line. AB - Using clonal cell cultures, a significant increase in chromosomal aberrations (aneuplolidy, dicentrics and chromatid breaks) were observed in MCF-7 cells compared with HeLa. BRCA1 expression was lower in MCF-7 cells than in HeLa cells. Since BRCA1 is known to play a role in the maintenance of chromosomal integrity, the increase in chromosomal aberrations in MCF-7 clones suggests that downregulation of BRCA1 expression could be one of the possible mechanisms for increased chromosomal instability in this cell line. PMID- 11407861 TI - Functional examination of microencapsulated bioengineered insulin-secreting beta cells. AB - Clonal insulin-secreting BRIN-BD11 cells engineered by electrofusion were encapsulated inside natrium alginate beads and cultured in RPMI 1640 culture media. Acute insulin secretory responses to glucose and amino acids were compared between microencapsulated cells and non-encapsulated cells maintained in monolayer culture. Encapsulated cells exhibited a 1.5-fold, 2.9-fold and 4.2-fold increase (P< 0.001) in insulin release in response to 16.7 mmol/l glucose, 10 mmol/l L-arginine and 10 mmol/l L-alanine respectively. Insulin output by non encapsulated cells was approximately 30% greater but the relative magnitudes of responses were similar. This is the first study to demonstrate the stability of cellular engineered insulin-secreting cells encapsulated in alginate beads, illustrating the utility of this approach for cellular engineering and potential transplantation in diabetes. PMID- 11407862 TI - Specific invasion of transformed cells by Escherichia coli A2 strain. AB - Bacteria of the spontaneously isolated non-pathogenic strain Escherichia coli A2 producing actin-specific protease ECP 32 (Usmanova and Khaitlina, 1989) were shown to be taken up by transformed cells, whereas finite and immortal cell lines were resistant to the infection. PMID- 11407863 TI - Effects of Th-2 type cytokines on human airway epithelial cells: interleukins-4, 5, and -13. AB - Proliferation and death of airway epithelial cells may be of importance in the pathogenesis of asthma. T-helper (Th)-2 response interleukins (IL)-4, -5, and -13 have the ability to induce proliferation of airway epithelial cells. PMID- 11407864 TI - Bifunctional effects of heparin-binding protein HBp17 on DNA synthesis in cells. AB - A 17 kD heparin-binding protein (HBp17) has a biphasic dose-dependent effect on DNA synthesis in 3T3 cells. Maximal stimulation of DNA synthesis occurs at 8 ng/ml HBp17, but a half-maximal inhibition occurs at approximately 500 ng/ml. This inhibition can easily be reversed by addition of 400 pg/ml aFGF or 100 pg/ml bFGF, whereas EGF had no effect. This biphasic action of HBp17 was also seen in human umbilical vein endothelial cells (HUVEC), whereas it was not found in the malignant cell line, A431-AJC. The functional relationship between HBp17 and FGF is discussed. PMID- 11407865 TI - The effectiveness of growth hormone, glutamine and a low-fat diet containing high carbohydrate on the enhancement of the function of remnant intestine among patients with short bowel syndrome: a review of published trials. AB - Many patients who undergo extensive resection of the gastrointestinal tract develop malabsorption which, in the worst cases, requires long-term parenteral nutrition at home, frequently on a permanent basis. Such patients can be defined as having intestinal failure resulting from the short bowel syndrome. In 1995, Wilmore's group hypothesized that the administration of growth factors and nutrients could enhance further adaptation of the remnant intestine and thereby improve absorption. The demonstrated, through controlled clinical trials, the benefit of such treatment among 47 adults with less than 200 cm of small intestine. Over the past few years, similar trials as well as animal experiments have been conducted by the same authors and other groups with conflicting results. We have performed a systematic search on the electronic databases for the purpose of identifying the evidence published so far on this subject. Our analysis suggests that the benefit of administering recombinant human growth hormone alone, or together with glutamine with or without a low-fat diet containing high-carbohydrate (fibre) is, if any, marginal. PMID- 11407866 TI - Clinical, social and rehabilitation status of long-term home parenteral nutrition patients: results of a European multicentre survey. AB - BACKGROUND: Home parenteral nutrition (HPN) is a lifesaving treatment in patients with intestinal failure. Dependency of nutritional support becomes permanent for the majority of patients who had received HPN for at least 2 years. The alternative to long term HPN in selected patients is intestinal transplantation. AIMS: To study some of the clinical, social and rehabilitation aspects of long term HPN treatment. METHODS: A survey was performed in nine European HPN centres. The questionnaire covered epidemiologic data, underlying diseases, intestinal anatomy, nutritional support and status, marital status, rehospitalization rate, HPN complications, rehabilitation score, drugs use, coexistent diseases and interest in intestinal transplantation. For some items, data were collected within 12 months prior to the evaluation. RESULTS: This survey included 228 patients with a median age of 49 years. The median duration of HPN was 7 years (range 2--24 years). Short bowel length less than 100 cm was reported in 65% of patients with a predominance of end-jejunostomy or jejuno-colonic anastomosis. Global subjective nutritional status was normal in 79% of the patients, who were supplied with a mean number of 5.6 bags of parenteral nutrition weekly. Rehospitalizations within the 12 months prior to evaluation accounted for a mean period of 23 days and were due to HPN complications in half of the cases. Catheter-related sepsis was the most frequent HPN-complication. Bone metabolism disorders, which seemed to be more common than liver diseases, were directly related to HPN duration. One-third of the HPN patients was regularly consuming analgesics or opiates. HPN impair complete rehabilitation status but may improve the status of patients who had a very low rehabilitation score before starting HPN. An interest of intestinal transplantation was noted in only 10% of medical teams and in 8% of HPN patients. CONCLUSIONS: This study is the largest European survey on long-term HPN patients with long standing or permanent intestinal failure. Data enlighten clinical, social and rehabilitation aspects of patients who could face the option of intestinal transplantation in the future. PMID- 11407867 TI - Effects of minor constituents (non-glyceride compounds) of virgin olive oil on plasma lipid concentrations in male Wistar rats. AB - We aimed to assess the effects of minor constituents (MC) from virgin olive oil upon the plasma lipid profile of experimental animals. Therefore, 32 Wistar rats were fed for 6 weeks with one of four different diets with a similar fatty acid composition but different MC: high-oleic sunflower oil (HOSO), virgin olive oil (VOO), 400%-MC enriched olive oil (EOO) and MC poor (impoverished) olive oil (IOO). At the end of the week 6 of dietary treatment, blood samples were obtained for analysis of lipid composition. A statistically significant influence was observed upon both total HDL (1.593+/-0.4, 1.204+/-0.212, 0.991+/-0.244 and 0.827+/-0.279 mmol/L for EOO, HOSO, VOO and IOO, respectively, Kruskal-Wallis test, P<0.05) and HDL(2)cholesterol levels (1.16+/-0.26, 0.576+/-0.191, 0.585+/ 0.216 and 0.583+/-0.207 mmol/L for EOO, HOSO, VOO and IOO, respectively, Kruskal Wallis test, P<0.05). No statistically significant effect was observed upon LDL cholesterol or triglycerides. Thus, MC supplementation has beneficial effects on HDL concentrations in Wistar rats. PMID- 11407868 TI - Assessment of nutritional status using biochemical and anthropometric variables in a nutritional intervention study of women with hip fracture. AB - BACKGROUND & AIMS: The aim of this study of women with hip fracture was to describe nutritional status with biochemical markers and anthropometric variables, and to evaluate the effect of nutritional intervention with the intention of increasing protein and energy intake. METHODS: The first consecutive 44 women were included, and used as controls. The next 44 were matched for age, fracture and mental state. Anthropometric variables, IGF-I, hormones and serum albumin were collected 4--6 days (baseline), 1 and 3 months after surgery. Twenty four women filled out a 7-day food record. RESULTS: At baseline, one fourth had BMI <20 kg/m(2)and subnormal triceps skinfold thickness. Baseline serum albumin, IGF-I and growth hormone levels were low, probably as an acute response to trauma. Women with BMI <20 kg/m(2)had lower IGF-I levels compared to those with higher BMI. At 3 months, one-third of both groups were protein and energy malnourished. The intervention group obtained higher daily energy percentage from fat but none of the groups reached their calculated energy need. CONCLUSIONS: Using biochemical markers in the acute postoperative situation to assess nutritional status is not recommended. The intervention had no impact on anthropometric or biochemical variables. PMID- 11407869 TI - Nutritional status after short-term dietary supplementation in hospitalized malnourished geriatric patients. AB - AIM: To examine the evolution of different parameters of the nutritional status after short-term oral protein-energy supplementation in moderately malnourished geriatric patients. METHODS: Seventeen hospitalized malnourished elderly patients and 12 healthy adults received dietary supplements for 10 days. A group of six malnourished elderly subjects served as controls. Spontaneous oral intakes, biological and biophysical markers of the nutritional status were measured. Fat free mass (FFM) was assessed using Dual energy X-ray absorptiometry (DXA), bio impedance analysis (BIA) and anthropometry. RESULTS: In elderly subjects, the supplementation significantly increased both dietary intake (energy +32%, protein +65%) and FFM (+1.3 kg, P<0.001) as assessed using DXA. BIA and anthropometric data correlated with DXA measurements in the elderly (BIA: r=0.68--0.80, anthropometry: r=0.80--0.89), but failed to reflect accurately the changes measured in FFM. Supplementation had no notable effect on biological markers in any of the groups. IGF-I and hand-grip strength were not significantly influenced by the supplementation despite trends towards an improvement. CONCLUSIONS: Monitoring short-term changes in nutritional status in malnourished elderly individuals is a problem in routine clinical management. Our data put in the limelight the changes in IGF-I values related to dietary supplementation, and, chiefly, suggest a prime role for the assessment of dietary intake and FFM, as assessed by DXA, as indicators of short-term efficacy of refeeding. Nevertheless larger studies are necessary to confirm the clinical and prognostic significance of the changes. PMID- 11407870 TI - Total energy expenditure in stable patients with cystic fibrosis. AB - BACKGROUND AND AIMS: Undernutrition is a common problem in patients with cystic fibrosis and is associated with a poor prognosis. The two aims of this study were to assess and compare the two main field techniques in the measurement of total energy expenditure and, secondly, to assess total energy expenditure in stable patients and compare with healthy controls. METHODS: Resting energy expenditure was measured using indirect calorimetry and total energy expenditure was measured using 24-h heart rate (HR) methodology and doubly isotopically labelled water. RESULTS: Seventeen patients, mean age 23 years and FEV(1)52% predicted and thirteen controls were recruited. Resting energy expenditure was higher in patients 0.24 (0.03) MJ/kg Fat-Free Mass (FFM) compared to controls 0.22 (0.02) MJ/kg FFM, P=0.02. Twenty-four hour heart rate underestimated total energy expenditure, 9.49 (1.85) MJ/day in patients compared to 11.69 (2.79) MJ/day using doubly labelled water. There was no difference in total energy expenditure in patients and controls using both methods, 11.69 (2.79) MJ/day compared to 11.38 (2.71) MJ/day using doubly isotopically labelled water. CONCLUSIONS: In clinically stable young adult patients with moderately severe respiratory disease total energy expenditure is comparable to that an a control population despite in increase in resting energy expenditure and both 24-h HR and doubly isotopically labelled water are suitable for use in patients with cystic fibrosis. PMID- 11407871 TI - Clinical assessment of HIV-associated lipodystrophy syndrome: bioelectrical impedance analysis, anthropometry and clinical scores. AB - BACKGROUND: Diagnosis of the HIV-associated lipodystrophy syndrome is based on clinical assessment, in lack of a consensus about case definition and reference methods. Three bedside methods were compared in their diagnostic value for lipodystrophy. PATIENTS AND METHODS: Consecutive HIV-infected outpatients (n=278) were investigated, 128 of which also had data from 1997 available. Segmental bioelectrical impedance analysis (BIA) and waist, hip and thigh circumferences were performed. Changes in seven body regions were rated by physicians and patients using linear analogue scale assessment (LASA). Diagnostic cut-off values were searched by receiver operator characteristics. RESULTS: Lipodystrophy was diagnosed in 85 patients (31%). BIA demonstrated higher fat-free mass in patients with lipodystrophy but not after controlling for body mass index and sex. Segmental BIA was not superior to whole body BIA in detecting lipodystrophy. Fat free mass increased from 1997 to 1999 independent from lipodystrophy. Waist-hip and waist-thigh ratios were higher in patients with lipodystrophy. BIA, anthropometry and LASA did not provide sufficient diagnostic cut-off values for lipodystrophy. Agreement between methods, and between patient and physician rating, was poor. CONCLUSION: These methods do not fulfil the urgent need for quantitative diagnostic tools for lipodystrophy. BIA estimates of fat free mass may be biased by lipodystrophy, indicating a need for re-calibration in HIV infected populations. PMID- 11407872 TI - Insulin-like growth factor 1 (IGF-1), a nutritional marker in patients with eating disorders. AB - BACKGROUND AND AIMS: Though low levels of insulin-like growth factor-1 (IGF-1) have been repeatedly reported in patients with eating disorders, the nutritional significance of IGF-1 has not been evaluated. The study aimed to assess the utility of IGF-1 for screening malnutrition and for monitoring nutrition intervention in patients with eating disorders. METHODS: IGF-1 and nutritional status were evaluated in 82 patients, 59 with anorexia nervosa (AN), and 23 with bulimia nervosa (BN). Nutritional assessment included the evaluation of body mass index (BMI), body fat (FAT) and muscle mass (MM), assessed by skinfold anthropometry, serum albumin, transthyretin and retinol-binding protein, energy and protein intake. IGF-1 and nutritional parameters were reevaluated in the early phase of refeeding (2-4 weeks) in 20 AN patients who entered a refeeding program. RESULTS: Mean IGF-1 z-score was -1.74+/-0.74 in AN, and -0.74+/-0.91 in BN. Serum proteins were reduced in only a minority of patients. IGF-1 correlated with BMI (r=0.64), FAT (r=0.57), MAMC (mid-arm muscle circumference) (r=0.58) and MM (r=0.66) (P<0.001), while it did not correlate with serum proteins. In the early phase of nutritional repletion serum proteins and anthropometric parameters did not vary significantly, while a prompt and marked increase (73.9%) of IGF-1 was observed. CONCLUSIONS: IGF-1 represents a biochemical marker of malnutrition and a sensitive index of nutritional repletion in patients with eating disorders. PMID- 11407873 TI - Nutritional deficits resulting from an almond-based infant diet. PMID- 11407875 TI - The meaning of hypoalbuminaemia in clinical practice. PMID- 11407876 TI - Causes and mechanisms of hypoalbuminaemia. PMID- 11407877 TI - The treatment of hypoalbuminaemia. PMID- 11407878 TI - Case discussion. Hypoalbuminaemia. PMID- 11407879 TI - Cost-containment with glutamine dipeptide-supplemented TPN. PMID- 11407881 TI - Leptosphaeria maculans, the causal agent of blackleg disease of Brassicas. AB - The loculoascomycete Leptosphaeria maculans (anamorph: Phoma lingam) causes blackleg of Brassicas, including Brassica napus (canola or rapeseed). This fungus probably comprises several morphologically similar species; taxonomic relationships between them are being clarified and nomenclature is being revised. The pathotype ("A" group) responsible for major economic losses to canola has been studied in more detail than other members of this species complex and is the focus of this review. L. maculans is haploid, outcrossing, can be transformed, and has a genome size of about 34 Mb. Preliminary genetic and physical maps have been developed and three genes involved in host specificity have been mapped. As yet, few genes have been characterized. Chemical analysis of fungal secondary metabolites has aided understanding of taxonomic relationships and of the host fungal interaction by the unraveling of pathways for detoxification of antimicrobial phytoalexins. Several phytotoxins (host and nonhost specific) have been identified and a complex pattern of regulation of their synthesis by fungal and host metabolites has been discovered. PMID- 11407882 TI - Cloning, expression, and characterization of the hxk-1 Gene from the white truffle Tuber borchii vittad.: A first step toward understanding sugar metabolism. AB - Recent biochemical investigations of Tuber borchii Vittad. mycelium have demonstrated the presence of three distinct forms of hexokinase (HK(M1), HK(M2), and HKM3). In the investigation described here, a gene coding for hexokinase (hxk 1) from T. borchii was isolated and characterized. The hxk-1 gene is characterized by an ORF of 1494 nucleotides and codes for a polypeptide of 497 aa. The gene was overexpressed in Escherichia coli, and the recombinant protein was kinetically characterized. The K(cat) value for fructose is in agreement with the data reported for the hexokinase of Yarrowia lipolytica, the Km for ATP is not dependent on the sugar used, and the enzyme is not inhibited by trehalose 6 phosphate or glucose 6-phosphate. The biochemical characteristics confirm that this enzyme is a hexokinase, as suggested by the Pileup results, and it corresponds to the HKM1 isoform. This work represents the first characterization of the key enzyme of the glycolytic pathway and the related gene in a Tuber species. PMID- 11407883 TI - Isolation and characterization of sexual sporulation mutants of Aspergillus nidulans. AB - For the genetic dissection of sexual sporulation in Aspergillus nidulans, we started a collection of ascosporeless mutants. After mutagenization of conidiospores with high doses of UV, we isolated 20 mutants with defects in ascospore formation. We crossed these mutants in two successive rounds with the wild-type strain. Eighteen of the 20 isolated mutants produced progeny with the original mutant phenotype in these crosses, and these mutants were further analyzed. All 18 analyzed mutations were recessive to wild type. We assigned them to 15 complementation groups, based on crosses between mutants. The mutants could be classified as follows according to their cytological phenotype: (1) no croziers, (2) arrest at prekaryogamy, (3) arrest in early meiotic prophase, (4) arrest in late meiotic prophase, (5) arrest in meiotic metaphase I, (6) defective postmeiotic mitosis and/or deliniation of ascospores, and (7) slow progression through the postmeiotic stages of ascospore formation. A large proportion of the mutants, namely 11 of 18, arrested in meiotic prophase or metaphase I. We discuss a possible approach for isolating the wild-type alleles of the genes that carry the sexual sporulation mutations. PMID- 11407884 TI - Characterization of pisatin-inducible cytochrome p450s in fungal pathogens of pea that detoxify the pea phytoalexin pisatin. AB - Many fungi that are pathogenic on pea have the ability to demethylate and thus detoxify the pea phytoalexin pisatin. This detoxification reaction has been studied most thoroughly in Nectria haematococca MP VI where it functions as a virulence trait. The enzyme catalyzing this reaction [pisatin demethylase (pda)] is a cytochrome P450. In the current study, the induction of whole-cell pda activity and the biochemical properties of pda in microsomal preparations from the pea pathogens Ascochyta pisi, Mycosphaerella pinodes, and Phoma pinodella are compared to the pda produced by N. haematococca. Based on cofactor requirements and their inhibition by carbon monoxide, cytochrome P450 inhibitors, and antibodies to NADPH:cytochrome P450 reductase, we conclude that the pdas from the other pea pathogens also are cytochrome P450s. All of the enzymes show a rather selective induction by pisatin, have a low K(m) toward pisatin, and have a fairly high degree of specificity toward pisatin as a substrate, suggesting that each pathogen may have a specific cytochrome P450 for detoxifying this plant antibiotic. Since the pdas in these fungi differ in their pattern of sensitivity to P450 inhibitors and display other minor biochemical differences, we suggest that these fungi may have independently evolved a specialized cytochrome P450 as a virulence trait for a common host. PMID- 11407885 TI - Elimination of active tad elements during the sexual phase of the Neurospora crassa life cycle. AB - Tad is an active LINE-like retrotransposon isolated from the Adiopodoume strain of Neurospora crassa. Extensive analysis of other Neurospora strains has revealed no other strain with active Tad, but all strains tested have multiple copies of defective Tad elements. We have examined the ability of Tad to survive during the sexual cycle of Neurospora and find that active Tad is rapidly eliminated. The characteristics of this elimination suggest that the repeat-induced point mutation (RIP) mechanism was responsible. By the use of transformation to switch the mating type of the Adiopodoume strain we concluded that this strain is not defective in the RIP process. Analysis of defective Tad elements isolated from a variety of strains indicates that the major difference between these elements and active Tad is due to the presence of a large number of G-C to A-T transition mutations. This would be expected if the changes were due primarily to the RIP process. Mapping of a selection of defective Tad elements reveals that they are present on all of the chromosomes; however, many of the elements are not widely shared among strains. This suggests that repeated introduction and elimination of Tad elements has occurred. Mechanisms that might be responsible for this repeated introduction are discussed. PMID- 11407886 TI - Isolation, characterization, and expression patterns of a DMC1 homolog from the basidiomycete Pleurotus ostreatus. AB - Here we describe the isolation of a Pleurotus ostreatus gene PoDMC1. The predicted amino acid sequence of the oyster mushroom gene is 62% identical to the yeast DMC1 and 60% identical to human DMC1. The highest degree of amino acid identity (88%), however, was shown with Coprinus CoLIM15, a DMC1 homolog recently found in Coprinus cinereus. The exact matching of sizes and positions of most introns in both basidiomycete genes underlines the close relationship between these DMC1 orthologs. The RecA homolog DMC1 from yeast and its orthologs from other species have been reported to be meiosis specific and essential for sporulation. Here we show that PoDMC1 is exclusively expressed in the lamellae/basidiospore fraction of fruit bodies and not in somatic cells of fruiting bodies or in vegetative mycelium. Furthermore, the gene is not expressed in the lamellae/basidiospore fraction of a nonsporulating mutant of P. ostreatus. Since one of the major problems in cultivating the oyster mushroom is the abundant sporulation that causes allergic reactions in man, PoDMC1 could be an important target gene in constructing sporeless Pleurotus strains. PMID- 11407887 TI - Evolutionary stability concepts for N-species frequency-dependent interactions. AB - The classical static concept of an evolutionarily stable strategy (ESS) for a single species gives rise to two new notions when there are more than two species (called an N-species ESS and RL-stability). The paper relates these to the dynamic stability of monomorphic and polymorphic evolutionary systems. It is shown that RL-stability implies the global asymptotic stability of either system with or without mutations. However, the N-species ESS only implies stability of the monomorphic system. PMID- 11407888 TI - Extinction and quasi-stationarity in the Verhulst logistic model. AB - We formulate and analyse a stochastic version of the Verhulst deterministic model for density-dependent growth of a single population. Three parameter regions with qualitatively different behaviours are identified. Explicit approximations of the quasi-stationary distribution and of the expected time to extinction are presented in each of these regions. The quasi-stationary distribution is approximately normal, and the time to extinction is long, in one of these regions. Another region has a short time to extinction and a quasi-stationary distribution that is approximately truncated geometric. A third region is a transition region between these two. Here the time to extinction is moderately long and the quasi-stationary distribution has a more complicated behaviour. Numerical illustrations are given. PMID- 11407889 TI - The size distribution of human hematogenous metastases. AB - The development of primary cancers and their subsequent metastases occur through a complex sequence of discrete steps. A hypothesis is proposed here whereby the time available for the growth of metastases is normally distributed, presumably as a consequence of the summation of multiple independently distributed time intervals from each of the steps and of the Central Limit Theorem. For exponentially growing metastases, the corresponding size distribution would be lognormal; Gompertzian growth would imply a modified (Gompertz-normal) distribution, where larger metastases would occur less frequently as a consequence of a decreased growth rate. These two size distributions were evaluated against 18 human autopsy cases where precise size measurements had been collected from over 3900 macroscopic hematogenous organ metastases. The lognormal distribution provided an approximate agreement. Its main deficiency was a tendency to over-represent metastases greater than 10 mm diameter. The Gompertz normal distribution provided more stringent agreement, correcting for this over representation. These observations supported the hypothesis of normally distributed growth times, and qualified the utility of the lognormal and Gompertz normal distributions for the size distribution of metastases. PMID- 11407890 TI - The biomechanics of an overarm throwing task: a simulation model examination of optimal timing of muscle activations. AB - A series of overarm throws, constrained to the parasagittal plane, were simulated using a muscle model actuated two-segment model representing the forearm and hand plus projectile. The parameters defining the modeled muscles and the anthropometry of the two-segment models were specific to the two young male subjects. All simulations commenced from a position of full elbow flexion and full wrist extension. The study was designed to elucidate the optimal inter muscular coordination strategies for throwing projectiles to achieve maximum range, as well as maximum projectile kinetic energy for a variety of projectile masses. A proximal to distal (PD) sequence of muscle activations was seen in many of the simulated throws but not all. Under certain conditions moment reversal produced a longer throw and greater projectile energy, and deactivation of the muscles resulted in increased projectile energy. Therefore, simple timing of muscle activation does not fully describe the patterns of muscle recruitment which can produce optimal throws. The models of the two subjects required different timings of muscle activations, and for some of the tasks used different coordination patterns. Optimal strategies were found to vary with the mass of the projectile, the anthropometry and the muscle characteristics of the subjects modeled. The tasks examined were relatively simple, but basic rules for coordinating these tasks were not evident. PMID- 11407891 TI - Relative safety factors against global buckling, anchorage rotation, and tissue rupture in wheat. AB - The objective of this study was to quantify the effect of specific physical and biological factors on the relative likelihood of biomechanical failure in wheat. Wind-related crop damage is a major obstacle to wheat production that costs several billion dollars per year. The four factors varied in this study were breeding line, dwarfing gene dose, soil type, and fertilization. A theoretical model describing the dynamic structural response of living plants was used to define margins of safety against global buckling, anchorage rotation, and tissue rupture. These relative safety factors were defined for each treatment in comparison with a tall wheat variety selected from a breeding line called Seri and grown in sandy soil with low fertilization. Compared to this reference, the relative safety factor against global buckling was increased (+39%, p<0.01), and the relative safety factor against anchorage rotation was decreased (-11%, p<0.025), by one allele of the dwarfing gene. The relative safety factor against tissue rupture was unaffected by the dwarfing gene but was consistently lower ( 26%, p<0.01) in a second breeding line called Kauz. Soil type and fertility did not affect the relative safety factors and this negative finding was significant at p<0.05. The key finding was that the strength of wheat was affected more by genetic rather than by environmental factors, which suggests that some varieties are intrinsically more robust than others. Also, the relative safety factor against anchorage rotation was inversely proportional to the relative safety factor against buckling, which suggests that there are competing constraints on the dynamic structural behavior of wheat. PMID- 11407892 TI - Molecular morphogenetic fields in the development of human dentition. AB - The mapping of the field of influence of specific regulatory molecules can provide a great deal of information on the molecular strategies that underlie the changes in the developmental program and macroevolutionary process. The strategy in this study was to use the variation in the number of teeth in the affected individuals of three mutant families with hypodontia, to determine the relative influence (relative molecular morphogenetic field) of MSX 1 and PAX 9 genes on the dental field. The variations in the pattern of symmetry of tooth agenesis were used in order to estimate the developmental stability of these genes. The approach used in the present work can help to explore new hypotheses linking development with the patterning of dentition during mammalian evolution. Furthermore, the developmental changes can be linked to changes in the molecular morphogenetic field of specific genes. PMID- 11407893 TI - Ombudsman or hotline for Gene therapy clinical trials? PMID- 11407894 TI - Gene transfer and athletics- an impending problem. PMID- 11407895 TI - Specific recognition of protein carboxy-terminal sequences by natural IgM antibodies in normal serum. AB - Our previous study indicated that normal serum contains complement-fixing natural IgM antibodies reacting with a large variety of randomly generated protein carboxy-termini. Here we show that the "carboxy-terminal" IgM (C-IgM) antibodies specifically react with short peptide sequences located immediately at the protein carboxy-terminus. The specificity of C-IgM-peptide interactions is tentatively defined by three to four amino acid residues. All carboxy-terminal peptides in a large peptide library apparently react with C-IgM antibodies. Immobilized synthetic peptides also react with C-IgM antibodies. No interaction of C-IgM antibodies with internal peptide sequences has been observed. C-IgM antibodies are present in germ-free and in athymic adult rats and are absent in newborn rats. The natural ubiquity of protein carboxy-termini in biological structures suggests that C-IgM could play an important role in antigen clearance and presentation to the immune system. From a practical viewpoint, the recognition of carboxy-terminal peptides by complement-fixing C-IgM antibodies has profound implications for the use of peptide- and protein-derivatized delivery vehicles and artificial materials. PMID- 11407896 TI - Nuclear translocation of lactosylated poly-L-lysine/cDNA complex in cystic fibrosis airway epithelial cells. AB - Poly-l-lysine, with 40% of its amino groups substituted with lactose, is an effective vector to transfer the CFTR gene into CF airway epithelial cells and correct the chloride channel dysfunction. The intracellular fate of the lactosylated poly-l-lysine/cDNA complex was studied using confocal microscopy. In the presence of chloroquine the complex remained intact during internalization, intracellular transport, and, most importantly, transport into the nucleus. When cells were transfected in the presence of agents that enhance transfection efficiency such as E5CA peptide, a fusogenic peptide, or glycerol a similar fate of the lactosylated poly-l-lysine/cDNA complex was seen. However, when these agents were omitted from the transfection medium, the complex remained in the perinuclear region. Uncomplexed lactosylated poly-l-lysine reached the nucleus efficiently. In contrast mannosylated poly-l-lysine or unsubstituted poly-l lysine complexed to plasmid did not. Therefore the nuclear accumulation of the complex may be attributed to the substitution of poly-l-lysine with lactose. It is hypothesized that the lactose residues provide for nuclear localization by means of targeting a potential lectin-like protein with galactose/lactose specificity. This mechanism may be responsible for the nuclear internalization of the complex. PMID- 11407897 TI - Regulation of angiogenesis and matrix remodeling by localized, matrix-mediated antisense gene delivery. AB - Implantation of biomaterials, such as glucose sensors, leads to the formation of a poorly vascularized collagenous capsule that can lead to implant failure. This process, known as the foreign body reaction (FBR), develops in response to almost all biomaterials and consists of overlapping phases similar to those in wound healing. Implantation of porous biomaterials, such as polyvinyl alcohol sponges, also leads to granuloma formation within the interstices of the sponge prior to encapsulation by the FBR. We asked whether delivery of an antisense cDNA for the potent angiogenesis inhibitor thrombospondin (TSP) 2 would enhance blood vessel formation and alter collagen fibrillogenesis in the sponge granuloma and capsule. Collagen solutions were mixed with plasmid to generate gene-activated matrices (GAMs) and applied to biomaterials that were then implanted subcutaneously. Sustained expression of plasmid-encoded proteins was observed at 2 weeks and a month following implantation. In vivo delivery of plasmids, encoding either sense or antisense TSP2 cDNA, altered blood vessel formation and collagen deposition in TSP2-null and wild-type mice, respectively. Untreated implants, implanted next to GAM-treated implants, did not show exogenous gene expression and did not elicit altered responses, suggesting that gene delivery was limited to implant sites. This method of antisense DNA delivery has the potential to improve the performance and life span of implantable delivery devices and biosensors. PMID- 11407898 TI - In vivo treatment of hemophilia A and mucopolysaccharidosis type VII using nonprimate lentiviral vectors. AB - Gene therapy holds great promise for the treatment of a variety of inherited diseases, including hemophilia A and mucopolysaccharidosis type VII (MPS VII). In both these disorders, subnormal levels of replacement protein have therapeutic effects. Thus we hypothesized that transduction of a small proportion of cells by feline immunodeficiency virus (FIV)-based lentiviral vectors might provide sufficient levels of transgene expression for phenotypic correction. We intravenously injected replication-deficient FIV-based vectors encoding either human factor VIII or human beta-glucuronidase into factor VIII-deficient or beta glucuronidase-deficient mice, respectively. This route of delivery targeted multiple organs, with the liver as the primary transduction site. In the hemophilia A mice, factor VIII expression persisted for the duration of the experiments (approximately 5 months), and recipient mice survived an otherwise lethal bleeding episode (tail-clipping). In mucopolysaccharidosis type VII mice, substantial beta-glucuronidase activity was detected in several tissues and corresponded with marked reduction of lysosomal storage in liver and spleen. These findings indicate that gene transfer with FIV-based lentiviral vectors can permanently introduce transgenes into a sufficient number of hepatocytes for long term therapeutic effect and suggest potential clinical value of FIV-based lentiviral vectors for treatment of hemophilia A and MPS VII. PMID- 11407899 TI - Human mesenchymal stem cells maintain transgene expression during expansion and differentiation. AB - Human adult bone marrow contains both hematopoietic stem cells that generate cells of all hematopoietic lineages and human mesenchymal stem cells (hMSCs), which support hematopoiesis and contribute to the regeneration of multiple connective tissues. The goal of the current study was to demonstrate that transduced hMSCs maintain transgene expression after stem cell differentiation in vitro and in vivo. We have introduced genes into cultured hMSCs by retroviral vector transfer and demonstrated long-term in vitro and in vivo expression of human interleukin 3 (hIL-3) and green fluorescent protein (GFP). Protocols were developed to achieve transduction efficiencies of 80-90% in these stem cells. In vitro expression of hIL-3 averaged 350 ng/10(6)cells/24 h over 17 passages (> 6 months) and GFP expression was stable over the same time period. Transduced hMSCs were able to differentiate into osteogenic, adipogenic, and chondrogenic lineages and maintained transgene expression after differentiation. Parallel studies were performed in vivo using NOD/SCID mice. Human MSCs expressing hIL-3 were cultured on several matrices and then delivered by subcutaneous, intravenous, and intraperitoneal routes. Sampling of peripheral blood demonstrated that systemic hIL-3 expression was maintained in the range of 100-800 pg/ml over a period of 3 months. These results illustrate the ability of hMSCs to express genes of therapeutic potential and demonstrate their potential clinical utility as cellular vehicles for systemic gene delivery. PMID- 11407900 TI - Tail-vein injection of mannan-binding lectin DNA leads to high expression levels of multimeric protein in liver. AB - The human plasma protein mannan-binding lectin (MBL) is an essential part of the innate immune defense system. Low levels of MBL are associated with recurrent infections and other clinically significant signs of a compromised immune defense. Previous studies have addressed the possibility of reconstitution therapy by the use of recombinant or plasma-derived protein. Natural MBL is a multimeric protein, which consists of up to 18 identical polypeptide chains. Synthesis by in vitro methods of MBL with the proper multimeric structure is difficult. We here report that mice obtain MBL levels comparable to those found in normal human plasma when injected with an MBL expression construct as naked plasmid DNA contained in a large volume of physiologic salt solution. The expression was confined to the liver and high MBL expression levels were obtained with less than 5% of the liver cells transfected. The multimeric structure of the MBL found in plasma of injected mice was similar to that of natural MBL. Thus, liver expression following injection of naked DNA is an alternative to reconstitution therapy with a protein having a complex quaternary structure. PMID- 11407901 TI - Primary adult human astrocytes as an ex vivo vehicle for beta-glucuronidase delivery in the brain. AB - Astrocytes are a good candidate cell type for brain transplantation: They are endogenous to the CNS, they have efficient secretory machinery, and they play a major role in neuronal support. We assessed the potential of genetically modified primary adult human astrocytes as vehicles for the delivery of secreted molecules in the mammalian CNS. We report that such cells can be efficiently transduced by a recombinant adenoviral vector carrying the human beta-glucuronidase cDNA (Ad/CMV*beta-glu) and that the transduced astrocytes produce large amounts of the enzyme. Released beta-glucuronidase could be captured, in vitro, by primary neurons and astrocytes and by a neuroblastoma cell line and beta-glucuronidase deficient fibroblasts. Following grafting into the mouse striatum, adult human astrocytes survived and expressed the transgene for at least 8 weeks. Moreover, the dosage of beta-glucuronidase activity within the grafted brains revealed high enzymatic levels at a long distance from the graft. These experiments document the grafting of engineered primary adult human astrocytes, allowing the release of a secreted therapeutic factor throughout the brain. PMID- 11407902 TI - Targeting of adenovirus to endothelial cells by a bispecific single-chain diabody directed against the adenovirus fiber knob domain and human endoglin (CD105). AB - The use of adenoviruses for antivascular cancer gene therapy is limited by their low transduction efficiency for endothelial cells. We have developed a recombinant bispecific antibody as a molecular bridge, linking the adenovirus capsid to the endothelial cell surface protein endoglin, for vascular targeting of adenoviruses. Endoglin (CD105), a component of the transforming growth factor beta receptor complex, represents a promising target for antivascular cancer therapy. Endoglin is expressed predominantly on endothelial cells and is upregulated in angiogenic areas of tumors. We isolated single-chain Fv fragments directed against human endoglin from a human semisynthetic antibody library. One of the isolated scFv fragments (scFv C4) bound specifically to various proliferating primary endothelial cells or cell lines including HUVEC, HDMEC, HMVEC, and HMEC. ScFv C4 was therefore used to construct a bispecific single chain diabody directed against endoglin and the adenovirus fiber knob domain (scDb EDG-Ad). This bispecific molecule mediated enhanced and selective adenovirus transduction of HUVECs, which was independent from binding to the coxsackievirus and adenovirus receptor (CAR) and alpha(v)-integrins. Thus, adenovirus infection was redirected to a new cellular receptor (CD105) and cell entry pathway. These results demonstrate the utility of bispecific single-chain diabodies, which can be produced in large quantities in bacteria, for the retargeting of adenoviruses in cancer gene therapy. PMID- 11407903 TI - Local high-capacity adenovirus-mediated mCTLA4Ig and mCD40Ig expression prolongs recombinant gene expression in skeletal muscle. AB - Multiple forms of muscular dystrophy are due to the absence of cytoskeletal muscle proteins that normally protect the integrity of muscle cells. The lack of any adequate treatments for these devastating diseases propels research toward the development of strategies for gene delivery to skeletal muscle. High-capacity adenoviral vectors (HC-AdV) devoid of all viral coding sequences have been developed to avoid expression of viral proteins by the gene therapy vector. However, the capsid proteins that are an essential component of the input viral vector and any residual helper virus in the vector preparation could induce an immune response. Furthermore, the therapeutic protein provided by a gene transfer vector presents the potential to induce an immune response in a patient who does not express a normal cellular protein due to genetic mutation. Therefore, we hypothesize that some immune suppression will be required with therapeutic gene delivery designed for the treatment of patients with inherited muscle diseases. In this study, we constructed and rescued three HC-AdVs expressing murine CTLA4Ig, murine CD40Ig, or both. The backbone vector without a gene insert was rescued as a negative control vector. The production of relevant proteins from each vector was determined in vitro. In vivo function of each of the immunosuppressant vectors was assayed by co-injection with an enhanced green fluorescent protein (EGFP)-expressing first-generation adenoviral vector (AdEGFP) into the tibialis anterior muscle of C57BL/10 mice. Higher levels of muscle EGFP expression were observed in animals receiving an immunosuppressant vector. Furthermore, the production of total anti-AdV and anti-EGFP antibodies was reduced in mice treated with each of the three immunosuppressant vectors. A second intramuscular administration of AdEGFP alone 4 weeks after the initial co injection was successful in all immunosuppressant vector-treated groups, but not in the negative control vector-treated group. All groups had a high antibody response to adenoviral proteins after the second injection of AdEGFP alone, indicating that the initial co-injection did not tolerize against vector capsid antigens. PMID- 11407904 TI - Gene transfer of p53 to arthritic joints stimulates synovial apoptosis and inhibits inflammation. AB - Rheumatoid arthritis (RA) is an autoimmune disease that primarily affects joints. During the pathogenesis of rheumatoid arthritis, the synovial lining becomes dramatically thickened and hyperplastic. This highly aggressive tissue invades and destroys articular cartilage and bone. Several lines of evidence suggest that the proliferation of the synovial tissue may be due to disruption in the control of the cell cycle or apoptotic pathways. In particular, mutations in the tumor suppressor protein p53 have been found in synovial tissue from RA joints. We have examined the effects of overexpression of p53 by adenoviral infection in synovial cells in culture and in synovial tissue in vivo in a rabbit model of arthritis. Here we demonstrate that p53 overexpression resulted in significant apoptosis in human and rabbit synovial cells in culture. Furthermore, intraarticular injection of Ad-p53 resulted in extensive and rapid induction of synovial apoptosis in the rabbit knee without affecting cartilage metabolism. Interestingly, a significant reduction in the leukocytic infiltrate was observed within 24 h postinfection of Ad.p53. These results suggest that intraarticular gene transfer of p53 is able to induce synovial apoptosis as well as reduce inflammation and thus may be useful clinically for the treatment of RA. PMID- 11407905 TI - Persistent low-level engraftment of rhesus peripheral blood progenitor cells transduced with the fanconi anemia C gene after conditioning with low-dose irradiation. AB - The hematopoietic stem cell has long been considered an ideal target for the introduction of therapeutic genes to treat human disorders such as Fanconi anemia (FA). Although recent progress in large animal models is encouraging, application to nonmalignant conditions is limited by the perceived necessity of myeloablative conditioning. We and others have shown that very low irradiation doses are sufficient to allow significant hematopoietic engraftment in murine hosts even after the introduction of xenogeneic genes. To determine the degree of engraftment of genetically modified cells attainable with very low irradiation doses in larger animals, we employed the rhesus macaque competitive repopulation model. Four animals underwent mobilization with stem cell factor (SCF) and granulocyte colony-stimulating factor (G-CSF) followed by apheresis. The apheresis product was enriched for the CD34-positive fraction by immunomagnetic selection and split equally for transduction with either G1FC26, a retroviral vector carrying the Fanconi anemia complementation group C gene, or PLII, a nonexpression control retroviral vector carrying both neomycin and beta galactosidase gene sequences modified to prevent translation. Transductions were performed daily in the presence of fresh IL-3, IL-6, SCF, and Flt-3 ligand on fibronectin-coated plates over 96 h. Animals were conditioned with a single dose of either 100 (n = 2) or 200 (n = 2) cGy and received the combined products of transduction on the following day. None of the animals experienced clinically significant neutropenia nor required the use of central line placement, transfusional support with blood products, or intravenous antibiotics. Using real time PCR, circulating levels of genetically modified cells as high as 1% were initially detected. Stable, albeit, significantly lower levels from both vector transduced aliquots (<0.1%) persisted beyond 12 months posttransplant in all four animals. Although not sufficient to correct the phenotype in many human disorders, stable low-level engraftment by genetically modified cells following low-intensity conditioning may prove adequate in disorders such as FA due to the selective advantage conferred upon corrected cells. PMID- 11407906 TI - Gene transfer into baboon repopulating cells: A comparison of Flt-3 Ligand and megakaryocyte growth and development factor versus IL-3 during ex vivo transduction. AB - Oncoretroviral vectors require division of target cells for successful transduction. In the case of hematopoietic repopulating cells this can be achieved by cytokine stimulation using growth factor combinations which facilitate gene transfer and maintain engraftment. Interleukin-3 (IL-3) has been widely used in growth factor combinations, although more recent data in the mouse showed reduced engraftment in the presence of IL-3. Here, we used a competitive repopulation assay to study the influence of IL-3 and the early acting cytokines megakaryocyte growth and development factor (MGDF) and Flt3-ligand (Flt3-L) on gene transfer efficiency during ex vivo transduction of hematopoietic repopulating cells. In a direct comparison, baboon CD34-enriched cells were transduced on CH-296 fibronectin fragment in the presence of either IL-6, stem cell factor (SCF), Flt3-L, and MGDF or IL-3, IL-6, and SCF. Animals were followed for up to 55 weeks, and analysis of peripheral blood leukocytes by semiquantitative polymerase chain reaction showed that both cytokine combinations achieved marking of repopulating cells. A trend toward increased gene marking, especially early after transplant (P = 0.06), was seen with the combination of IL 6, SCF, Flt3-L, and MGDF. However, the highest gene marking was achieved when IL 3 was combined with early acting cytokines, suggesting that the difference observed in this study was probably due to the addition of MGDF and Flt3-L and not due to a negative effect of IL-3 on engraftment. PMID- 11407907 TI - Defective lentiviral vectors are efficiently trafficked by HIV-1 and inhibit its replication. AB - Gene therapy against HIV infection should involve vector-mediated delivery of anti-HIV therapeutic genes into T-lymphocytes and macrophages or, alternatively, hematopoietic progenitors. Transduction of mature cells with defective vectors would have limited success because the vector would disappear with cell turnover. However, if a vector could be trafficked by wild-type HIV, initial transduction of a majority of the population would not be required, as the vector would be able to spread. We describe HIV-1-based lentiviral vectors that are efficiently packaged and trafficked by HIV-1, allowing a small number of cells initially transduced to spread the vector within a nontransduced cell population. We examined whether the presence or absence of the rev gene and the Rev-responsive element (RRE) would have a noticeable effect on the ability of lentiviral vectors to be trafficked and to inhibit HIV-1 replication. We found that replacement of rev/RRE with a constitutive transport element from Mason-Pfizer monkey virus had no apparent effect on trafficking and did not change the intrinsic inhibitory abilities of the vectors. We also constructed a rev/RRE-independent HIV-1-derived vector carrying a trans-dominant negative mutant of HIV-1 Rev, RevM10. This vector was less efficiently trafficked by HIV-1 and, despite the presence of an anti-HIV-1 gene, RevM10, was less efficient at inhibiting HIV-1 replication when introduced into a target T-cell population. PMID- 11407908 TI - Adeno-associated virus 2-mediated transduction and erythroid lineage-restricted long-term expression of the human beta-globin gene in hematopoietic cells from homozygous beta-thalassemic mice. AB - Adeno-associated virus 2 (AAV), a nonpathogenic human parvovirus, has gained attention as a potentially useful vector for human gene therapy. Here, we report successful AAV-mediated stable transduction and high-efficiency, long-term, erythroid lineage-restricted expression of a human beta-globin gene in primary murine hematopoietic stem cells in vivo. Bone marrow-derived primitive Sca-1(+), lin(-) hematopoietic stem cells from homozygous beta-thalassemic mice were transduced ex vivo with a recombinant AAV vector containing a normal human beta globin gene followed by transplantation into low-dose-irradiated B6.c-kitW(41/41) anemic recipient mice. Six months posttransplantation, tail-vein blood samples were analyzed by PCR amplification to document the presence of the transduced human beta-globin gene sequences in the peripheral blood cells. Semiquantitative PCR analyses revealed that the transduced human beta-globin gene sequences were present at approximately 1 copy per cell. The efficiency of the human beta-globin gene expression was determined to be up to 35% compared with the murine endogenous beta-globin gene by semiquantitative RT-PCR analyses. Peripheral blood samples from several positive recipient mice obtained 10 months posttransplantation were fractionated to obtain enriched populations of granulocytes, lymphocytes, and erythroid cells. PCR analyses revealed the presence of the human beta-globin gene sequences in granulocytes and lymphocytes, indicating multilineage reconstitution. However, only the erythroid population was positive following RT-PCR analyses, suggesting lineage-restricted expression of the transduced human beta-globin gene. Southern blot analyses of total genomic DNA samples isolated from bone marrow cells from transplanted mice also documented proviral integration. These results provide further support for the potential use of recombinant AAV vectors in gene therapy of beta-thalassemia and sickle-cell disease. PMID- 11407909 TI - Long-term and therapeutic-level hepatic gene expression of human factor IX after naked plasmid transfer in vivo. AB - Naked DNA transfer of a high-expressing human factor IX (hFIX) plasmid yielded long-term (over 1 1/2 years) and therapeutic-level (0.5-2 microg/ml) gene expression of hFIX from mouse livers. The expression cassette contained a hepatic locus control region from the ApoE gene locus, an alpha1-anti-trypsin promoter, hFIX cDNA, a portion of the hFIX first intron, and a bovine growth hormone polyadenylation signal. In contrast, a hFIX plasmid containing the expression cassette without effective regulatory elements produced initially low-level gene expression that rapidly declined to undetectable levels. Southern analyses of the cellular DNA indicated that the majority of the input genome from either vector persisted as episomal forms of the original plasmids. Together with RT-PCR analyses of the transcripts, these data indicated that at least two processes are critical for sustained gene expression: persistence of vector DNA and transcriptional/posttranscriptional activation. Liver regeneration after partial hepatectomy resulted in a significant decline in transgene expression, further suggestive of decreased episomal plasmid maintenance rather than transgene integration. Transaminase levels and liver histology showed that rapid intravenous plasmid injection into mice induced transient focal acute liver damage (< 5% of hepatocytes), which was rapidly repaired within 3 to 10 days and resulted thereafter in histologically normal tissue. No significant differences were observed between rapid injection of plasmid and saline control solutions. Transient, very low level antibodies directed against hFIX did not prevent the circulation of therapeutic levels of the protein. Gene transfer of hFIX plasmid DNA into liver elicited neither transgene-specific cytotoxic effect nor long-term toxicity. These results demonstrate that long-term expression of hFIX can be achieved by nonviral plasmid transfer and suggest that this occurs independent of integration. PMID- 11407910 TI - HSV amplicon-mediated neurotrophin-3 expression protects murine spiral ganglion neurons from cisplatin-induced damage. AB - Ototoxicity is a major dose-limiting side effect of cisplatin (DDP) administration due to its propensity to induce destruction of hair cells and neurons in the auditory system. Previous studies demonstrated that TrkC expressing spiral ganglion neurons (SGN) are protected from the cytotoxic effects of DDP by localized delivery of the trophic factor neurotrophin-3 (NT-3). Successful in vivo implementation of such a therapy requires the development of an efficient gene delivery vehicle for expression of NT-3 within the cochlea. To this end, we constructed a herpes simplex virus (HSV) amplicon vector that expressed a c-Myc-tagged NT-3 chimera (HSVnt-3myc). Helper virus-free vector stocks were initially evaluated in vitro for their capacity to direct expression of NT-3 mRNA and protein. Transduction of cultured murine cochlear explants with HSVnt-3myc resulted in production of NT-3 mRNA and protein up to 3 ng/ml as measured over a 48-h period in culture supernatants. To determine whether NT-3 overexpression could abrogate DDP toxicity, cochlear explants were transduced with HSVnt-3myc or a murine intestinal alkaline phosphatase-expressing control vector, HSVmiap, and then exposed to cisplatin. HSVnt-3myc-transduced cochlear explants harbored significantly greater numbers of surviving SGNs than those infected with control virus. These data demonstrate that amplicon-mediated NT-3 transduction can attenuate the ototoxic action of DDP on organotypic culture. The potency of NT-3 in protecting spiral ganglion neurons from degeneration suggests that in vivo neurotrophin-based gene therapy may be useful for the prevention and/or treatment of hearing disorders. PMID- 11407911 TI - Incorporation of tumor-targeting peptides into recombinant adeno-associated virus capsids. AB - The human parvovirus adeno-associated virus type 2 (AAV-2) possesses many features that make it an attractive vector for gene delivery in vivo. However, its broad host range may limit its usefulness and effectivity in several gene therapy applications in which transgene expression needs to be limited to a specific organ or cell type. In this study, we explored the possibility of directing recombinant AAV-2 transduction by incorporating targeting peptides previously isolated by in vivo phage display. Two putative loops within the AAV-2 capsid were examined as sites for incorporation of peptides. We tested the effects of deleting these loops and different strategies for the incorporation of several targeting peptides. The tumor-targeting sequence NGRAHA and a Myc epitope control were incorporated either as insertions or as replacements of the original capsid sequence. Viruses were assessed for packaging, accessibility of incorporated peptides, heparin binding, and transduction in a range of cell lines. Whereas recombinant viruses containing mutant capsid proteins were produced efficiently, transduction of several cell lines was significantly impaired for most modifications. However, certain mutants containing the peptide motif NGR, which binds CD13 (a receptor expressed in angiogenic vasculature and in many tumor cell lines), displayed an altered tropism toward cells expressing this receptor. Based on this work and previous studies, possible strategies for achieving in vivo targeting of recombinant AAV-2 are discussed. PMID- 11407912 TI - Development of a competitive PCR method for physical titration of recombinant EBV vector in a helper-dependent packaging system. AB - Epstein-Barr virus (EBV) is a gamma-herpesvirus with B lymphotropism and a double stranded DNA genome of 172 kb that is episomally maintained in permissive cells during latency. EBV-based vectors containing minimal cis elements for replication, amplification, and helper-dependent packaging in a producer cell line HH514 have been developed to deliver therapeutic/suicide transgenes as infectious viral particles (miniEBV) to EBV-transformed B lymphoblastoid cells or B lymphoma cells. A quantitative, competitive PCR-based assay was developed to determine the relative packaging efficiencies of miniEBV and helper P3HR1 coproduced in HH514 cells. This provides a rapid and accurate quantitation of the physical titer of the virus preparation, which helps preserve the biological titer of the virus preparation and increase the efficiency of transgene delivery by miniEBV infection. In addition, it provides a sensitive and accurate way to evaluate future development of a helper-free packaging system by detecting any possible helper virus contamination. PMID- 11407913 TI - Isolation and characterization of a Streptococcus thermophilus plasmid closely related to the pMV158 family. AB - Twenty-two Streptococcus thermophilus strains used for milk fermentations were analyzed for their plasmid content and 13 of them (59%) were found to contain one or two plasmids. Fifteen S. thermophilus plasmids were divided into four groups using DNA homology. Ten plasmids were classified within group A and they shared homologies with all the previously sequenced S. thermophilus plasmids. Three plasmids (group B) hybridized with each other and two plasmids only hybridized with themselves (groups C and D). Single-stranded DNA was detected within strains containing plasmids of groups A, C, and D, indicating that they replicate via a rolling-circle mode. The only plasmid of group C, named pSMQ172, was further characterized. This 4230-bp plasmid replicates in Escherichia coli, Lactococcus lactis, and Streptococcus salivarius and does not confer phage resistance. Comparisons with databases showed that pSMQ172 was related to pMV158 of Streptococcus agalactiae and to pSSU1 of Streptococcus suis. These results suggest that genetic exchanges may have occurred between pathogenic and nonpathogenic streptococci. PMID- 11407914 TI - Genetic organization of plasmid pXF51 from the plant pathogen Xylella fastidiosa. AB - The sequence of plasmid pXF51 from the plant pathogen Xylella fastidiosa, the causal agent of citrus variegated chlorosis, has been analyzed. This plasmid codes for 65 open reading frames (ORFs), organized into four main regions, containing genes related to replication, mobilization, and conjugative transfer. Twenty-five ORFs have no counterparts in the public sequence databases, and 7 are similar to conserved hypothetical proteins from other bacteria. A pXF51 incompatibility group has not been determined, as we could not find a typical replication origin. One cluster of conjugation-related genes (trb) seems to be incomplete in pXF51, and a copy of this sequence is found in the chromosome, suggesting it was generated by a duplication event. A second cluster (tra) contains all genes necessary for conjugation transfer to occur, showing a conserved organization with other conjugative plasmids. An identifiable origin of transfer similar to oriT from IncP plasmids is found adjacent to genes encoding two mobilization proteins. None of the ORFs with putative assigned function could be predicted as having a role in pathogenesis, except for a virulence-associated protein D homolog. These results indicate that even though pXF51 appears not to have a direct role in Xylella pathogenesis, it is a conjugative plasmid that could be important for lateral gene transfer in this bacterium. This property may be of great importance for future development of transformation techniques in X. fastidiosa. PMID- 11407915 TI - P1 and NR1 plasmid replication during the cell cycle of Escherichia coli. AB - Replication patterns of the miniP1 plasmid pZC176, the miniNR1 plasmid pRR933, and the high-copy miniNR1 derivative pRR942 were examined during the Escherichia coli cell division cycle and compared to the cycle-specific replication pattern of a minichromosome and the cycle nonspecific pattern of pBR322. In E. coli cells growing with doubling times of 40 and 60 min, the miniP1 plasmid was found to replicate with a slight periodicity during the division cycle. The periodicity was not nearly as pronounced as that of the minichromosome, was not affected by the presence of a minichromosome, and was not evident in cells growing more rapidly with a doubling time of 25 min. Both miniNR1 plasmids, pRR933 and pRR942, replicated with patterns indistinguishable from that of pBR322 and clearly different from that of the minichromosome. It is concluded that both P1 and NR1 plasmids can replicate at all stages of the cell cycle but that P1 displays a slight periodicity in replication probability in the cycle of slower growing cells. This periodicity does not appear to be coupled to a specific age in the cycle, but could be associated with the achievement of a specific cell mass per plasmid. During temperature shifts of a dnaC(Ts) mutant, the miniP1 plasmid and pBR322 replicated with similar patterns that differed from that of the minichromosome, but were consistent with a brief eclipse between rounds of replication. PMID- 11407916 TI - Molecular characterization of the klebicin B plasmid of Klebsiella pneumoniae. AB - The nucleotide sequence of a bacteriocin-encoding plasmid isolated from Klebsiella pneumoniae (pKlebB-K17/80) has been determined. The encoded klebicin B protein is similar in sequence to the DNase pyocins and colicins, suggesting that klebicin B functions as a nonspecific endonuclease. The klebicin gene cluster, as well as the plasmid backbone, is a chimera, with regions similar to those of pore former colicins, nuclease pyocins and colicins as well as noncolicinogenic plasmids. Similarities between pKlebB plasmid maintenance functions and those of the colicin E1 plasmid suggest that pKlebB is a member of the ColE1 plasmid replication family. PMID- 11407917 TI - Characterization and sequence analysis of the replicator region of the novel plasmid pALC1 from Paracoccus alcaliphilus. AB - The replicator region of a low-copy-number plasmid, pALC1, of Paracoccus alcaliphilus JCM 7364 was cloned in a form of the minireplicon pALC100 (3.6 kb). The host range of the minireplicon embraces several species of genus Paracoccus, as well as Agrobacterium tumefaciens, Rhizobium leguminosarum, and Rhodobacter sphaeroides (all belonging to alpha-Proteobacteria), but not Escherichia coli. The complete nucleotide sequence of the replicator region (2276 bp) revealed the presence of one complete open reading frame coding for the 28.4-kDa protein (RepA) with similarity to replication proteins of plasmid pSW500 of Erwinia stewartii and pVS1 of Pseudomonas fluorescens. The iteron-like region was identified upstream of the repA gene and consisted of two clusters of repeated sequences (17 bp long) separated by a putative DnaA box. Analysis of the predicted amino acid sequence of two adjacent incomplete ORFs suggests the localization of repA between genes involved in conjugation (traG) and partitioning (parA) within the pALC1 genome. PMID- 11407918 TI - Sequence analysis of the plasmid pRRI2 from the rumen bacterium Prevotella ruminicola 223/M2/7 and the use of pRRI2 in Prevotella/Bacteroides Shuttle Vectors. AB - pRRI2 is a small cryptic plasmid from the rumen bacterium Prevotella ruminicola 223/M2/7 which has been used for the construction of shuttle vectors (pRH3 and pRRI207) that replicate in many Bacteroides/Prevotella strains as well as in Escherichia coli. Sequence analysis of pRRI2 reveals that it is a 3240-bp plasmid carrying two clear open reading frames. Rep, encoded by ORF1, shows 48 and 47% amino acid sequence identity with RepA proteins from Bacteroides vulgatus and Bacteroides fragilis, respectively. ORF2, named Pre, shares 34% amino acid sequence identity with a putative plasmid recombination protein from the Flavobacterium spp. plasmid pFL1 and 30% amino acid sequence identity with BmpH from B. fragilis Tn5520. Disruption of ORF1 with HindIII prevents replication and maintenance in Bacteroides spp. hosts, but shuttle vectors carrying pRRI2 interrupted within ORF2, by EcoRI*, are able to replicate. pRRI2 shows no significant similarity with the only other P. ruminicola plasmid to have been studied previously, pRAM4. PMID- 11407919 TI - Isolation and characterization of group II introns from Pseudomonas alcaligenes and Pseudomonas putida. AB - Group II introns isolated from Pseudomonas alcaligenes NCIB 9867, Pseudomonas putida NCIB 9869, and P. putida KT2440 were closely related with nucleotide sequence identities of between 87 and 96%. The genome of P. alcaligenes also harbored a truncated group II intron of 682 bp that lacks the gene for the intron encoded protein (IEP). Unlike most bacterial group II introns, the Pseudomonas introns were found to lack the Zn domains in their IEPs, did not appear to interrupt any genes, and were located downstream of open reading frames which were adjacent to hairpin loop structures that resemble rho-independent terminators. These structures also contain the intron binding sites 1 and 2 (IBS1 and IBS2 sequences) that were required for intron target site recognition in transposition. One of the group II introns found in P. alcaligenes, Xln3, was shown to have transposed from the chromosome to the endogenous pRA2 plasmid at a site adjacent to IBS1- and IBS2-like sequences. PMID- 11407930 TI - Nausea threshold in apparently healthy individuals who drink fluids containing graded concentrations of copper. AB - Ingestion of drinking water with a high copper content may induce acute gastrointestinal effects, mainly nausea and vomiting, rarely diarrhea and abdominal pain. The objectives of this study were to define nausea threshold in apparently healthy adult volunteers who received graded concentrations of copper and to explore how individual thresholds were modified by delivering copper in an orange-flavored drink. Sixty-one healthy subjects received 200 mL of a copper containing solution in purified water, at concentrations 0, 2, 4, 6, 8, 10, and 12 mg/L, as copper sulfate, in random order. Nausea threshold concentration for first response was established and then this threshold was confirmed. Subsequently, following the same design, subjects received the same copper concentrations (up to 12 mg/L), delivered in an orange-flavored drink, starting at the confirmed threshold concentration found in water. Mild nausea shortly after ingestion of copper-containing water was the most frequent finding (33/61 subjects), starting at 4 mg/L; vomiting was observed in 7 individuals, starting at 6 mg/L. The NOEL for copper in purified water was 2 and 4 mg/L for nausea and vomiting, respectively. When copper was provided as an orange-flavored drink, 11 subjects (18%) reported nausea, starting at 8 mg Cu/L, and no subjects vomited up to 12 mg Cu/L. It is concluded that after consumption of copper in purified water, the NOEL is 2 mg Cu/L and the LOAEL 4 mg Cu/L for nausea, while tolerable intake is between 2 and 4 mg Cu/L in water depending on whether apparent or confirmed nausea is used as the criterion to define critical effects. PMID- 11407931 TI - Effects of low levels of ciprofloxacin on a chemostat model of the human colonic microflora. AB - To study the utility of an in vitro model system for assessing the effect of low concentrations of a fluoroquinolone (FQ) drug on the ecology of the human intestinal microflora, chemostats containing human fecal flora were exposed to 0.43, 4.3, and 43microg of ciprofloxacin (CI) per milliliter. Prior to and during drug exposure, we assayed short-chain fatty acids (SCFA), bacterial populations, and the relative levels of susceptibility of these populations to CI and trovafloxacin (TV), a newer related FQ with increased activity against anaerobes. The degree to which CI affected the chemostat ecology was measured statistically by comparing observed data with the corresponding predicted "no effect" level. No changes in total SCFA were observed; only butyrate was significantly higher at the intermediate and high-dose levels. Enterococci counts and the levels of susceptibility to CI among enterococci were also unaffected. Escherichia coli counts decreased in a dose-dependent manner. Susceptibility levels in E. coli followed no interpretable pattern. Bacteroides fragilis group (BfG) counts decreased significantly following exposure to 43 and 4.3microg/mL CI. Ciprofloxacin susceptibility among the BfG in these chemostats was not determined because the BfG counts were too low (less than 30 colonies per plate) when undiluted chemostat samples were plated. However, within 2 days of exposure to 0.43microg/mL CI, the percentage of BfG resistant to 4microg/mL CI increased to over 95%. Before exposure, all BfG were susceptible to both CI (2microg/mL) and TV (0.25microg/mL). All BfG isolated during exposure were resistant to both CI (4microg/mL) and TV (2microg/mL). Resistance selection in the BfG was unexpected as the MIC(90) of CI for B. fragilis is 8microg/mL. Since the average colon flora is about 20% B. fragilis and other bacteroides, CI may impact the human gut flora even at subtherapeutic levels. PMID- 11407932 TI - Benefit and risk of organic ultraviolet filters. AB - Modern sunscreen products provide broad-spectrum UV protection and may contain one or several UV filters. A modern UV filter should be heat and photostable, water resistant, nontoxic, and easy to formulate. Identification of a substance that meets these criteria is as difficult as discovering a new drug; hundreds of new molecules are synthesized and screened before a lead candidate is identified. The most important aspect in the development of a new UV filter is its safety. In our laboratories, the safety of new ultraviolet filters is assessed by an initial in vitro screen including photostability, cytotoxicity, photocytotoxicity, genotoxicity, and photogenotoxicity tests. These tests are performed in mammalian, yeast, and bacterial cell systems. Skin penetration potential is measured in vitro using human skin or, when required by regulations, in vivo. Because modern sunscreens are selected on the basis of their retention on and in the stratum corneum and are formulated as poorly penetrating emulsions, they generally have very low to negligible penetration rates. The safety and efficacy of UV filters are regulated and approved by national and international health authorities. Safety standards in the European Union, United States, or Japan stipulate that new filters pass a stringent toxicological safety evaluation prior to approval. The safety dossier of a new UV filter resembles that of a new drug and includes acute toxicity, irritation, sensitization, phototoxicity, photosensitization, subchronic and chronic toxicity, reproductive toxicity, genotoxicity, photogenotoxicity, carcinogenicity, and, in the United States, photocarcinogenicity testing. The margin of safety of new UV filters for application to humans is estimated by comparing the potential human systemic exposure with the no-effect level from in vivo toxicity studies. Only substances with a safe toxicological profile and a margin of safety of at least 100-fold are approved for human use. Finally, prior to marketing, new UV filters undergo stringent human testing to confirm their efficacy as well as the absence of irritation, sensitization, photoirritation, and photosensitization potential in man. UV filters not only protect against acute skin injury, such as sunburn, but also against long-term and chronic skin damage, including cellular DNA damage, photoinduced immune suppression, and, by extension, skin cancer. The protection provided by modern sunscreens against UV-induced skin cancer was shown in animal photocarcinogenicity studies and confirmed by numerous in vitro, animal, and human investigations: UV filters protect the p53 tumor suppressor gene from damage and prevent UV-induced immune suppression. Recent studies suggest that sunscreens protect against precursor lesions of skin cancer, such as actinic keratoses. Additional benefits of ultraviolet filters include prevention of photodermatoses, such as polymorphic light eruption, and, possibly, photoaging. Modern sunscreens are safe for children and adults. Percutaneous penetration and irritation rates of topically applied substances in children and adults are similar. The principal protective measure is to keep children out of the sun and/or to cover them with protective clothes; however, sunscreens are a safe and effective and often the only feasible defense of children against UV radiation. In conclusion, sunscreens are safe protective devices that undergo stringent safety and efficacy evaluation. PMID- 11407933 TI - PCBs and neurodevelopmental effects in Michigan children: an evaluation of exposure and dose characterization. AB - Despite the fact that PCB levels in the general environment have continued to decline over the past decade, concern for potential neurodevelopmental deficits from in utero exposure to these compounds remains unabated. In fact, some regulatory and scientific bodies have concluded that the evidence suggesting that prenatal exposure to PCBs may lead to neurodevelopmental deficits is one of the greatest public health concerns surrounding PCBs. The primary basis for the concern that low-level in utero exposure to PCBs causes neurodevelopmental deficits in children is a series of reports on a cohort of Michigan children presumably exposed to PCBs as a result of their mother's consumption of Great Lakes fish. These children, known collectively as the Jacobson cohort, have been followed from birth to 11 years of age. The investigators following these children concluded that they have demonstrated persistent neurodevelopmental effects in this cohort attributable solely to PCBs. However, a detailed analysis of the cohort's exposure characterization, particularly in the initial reports, reveals considerable uncertainty as to the actual exposure status of mothers characterized as "fish eaters" and their offspring. Failure to adequately characterize the PCB exposure of these mothers, or their children, precludes any causal association between in utero exposure to PCBs and neurodevelopmental deficits. PMID- 11407934 TI - Chemical categories for health hazard identification: a feasibility study. AB - The use of chemical categories has been suggested in order to lower the number of chemicals tested in the High Production Volume (HPV) Chemical Challenge Program. In this investigation we examined the reliability of using organic chemical categories to classify chemicals as either toxic or nontoxic for individual toxicological effects as well as for panels of such endpoints. The analyses indicate that chemical categories are unable to consistently identify groups of chemicals with similar toxic responses either for a multiplicity of endpoints or for single effects. Our analyses suggest that if chemical categories are to be used to identify health hazards, that computer-based SAR approaches appear to be superior to arbitrary chemical categories for predicting specific toxicological effects but they are not, at this time, useful for defining the overall toxicity. PMID- 11407935 TI - Interspecies comparisons of the toxicity of asbestos and synthetic vitreous fibers: a weight-of-the-evidence approach. AB - This analysis reviews the available literature on interspecies comparisons of the toxicity of asbestos and synthetic vitreous fibers (SVFs). This topic is of substantial practical importance because most quantitative risk analyses on the effects of inhalation of SVFs are based upon extrapolation of data from rodent inhalation studies. Available information on interspecies comparisons for both dosimetry (the relation between exposure concentration and fiber lung burden) and potency (the relation between lung burden and disease) is summarized. Dosimetry models indicate that, on a normalized basis, fiber deposition and clearance rates are lower in humans than rats. Potency is less well understood than dosimetry, in part because the source of relevant human data is asbestos studies, which are adequate to demonstrate hazard, but are problematic in other regards. There are significant interspecies differences between the mouse, hamster, rat, and human. The available evidence suggests that the rat is preferable as a model for the human. Rats develop fibrosis at comparable lung burdens [10(6) long (> 20 microm length) fibers per gram of dry lung] to those in humans. This analysis concludes that, on a weight-of-evidence basis, there is no reason to conclude that humans are more sensitive to fibers than rats with respect to the development of lung cancer. PMID- 11407936 TI - Chromate concentration bias in primer paint particles. AB - Chromate-containing primer paints are used to prevent corrosion on metal surfaces. Chromate contains hexavalent chromium (Cr6+), a human carcinogen. The objective of this research was to determine if there is a bias in the fraction of chromate found in various particle sizes generated during primer painting operations. A solvent-based, aviation primer paint was sprayed using a high volume, low-pressure spray gun. Paint particles were collected and separated by size with seven-stage cascade impactors. It was determined that particles with a mass aerodynamic diameter < 2.0 microm contained significantly less Cr6+ per dry weight of paint than particles > 2.0 microm (P < 0.001). The median concentration of Cr6+ in particles < 2.0 microm is 18 micro g of Cr/mg of dry paint and the median concentration for particles > 2.0 microm is 70 microg of Cr/mg of dry paint. The mixed paint contains 18.75% strontium chromate, which equates to a ratio of 67 microg of Cr/mg of dry paint. Particles > 2.0 microm are more likely to impact in the upper tracheobronchial regions of the lung where mucociliary clearance is relatively rapid. Additionally, chromate emissions from spraying operations may be overestimated because larger particles, which are more easily trapped on an air filter, contain more chromate than the smaller particles, which are more likely to bypass an air filter. PMID- 11407937 TI - The power of the European Union protocol to test for carcinogenicity of inhaled fibers. AB - We evaluate the power of a recent protocol proposed by the European Union (EU) for testing the carcinogenicity of inhaled fibers. We assume that every fiber has oncogenic potential determined by its biopersistence. We use a recently estimated potency for the oncogenic potential of fibers together with experimentally determined "weighted" half-lives (WHL) of a dozen fibers to generate simulations of long-term bioassays conducted according to the EU protocol. We analyze these experiments using standard statistical techniques and determine the number of tests that would have yielded significant results. We conclude that the EU protocol will readily detect the carcinogenic effect of long-lived fibers, such as amosite with a WHL of more than 450 days, and usually detect the effect of fibers, such as RCF1a, with WHL of about 40-50 days. However, the EU protocol has very low power to detect effects of short-lived fibers, such as X607, with WHL of about 10 days. PMID- 11407938 TI - Safety assessment of DHA-rich microalgae from Schizochytrium sp. AB - Schizochytrium sp. dried microalgae (DRM) contains oil rich in highly unsaturated fatty acids (PUFAs). Docosahexaenoic acid (DHA n-3) is the most abundant PUFA component of the oil. DHA-rich oil extracted from Schizochytrium sp. is intended for use as a nutritional ingredient in foods. As part of a comprehensive safety assessment program, the reproductive toxicity of DRM was examined in Sprague Dawley-derived rats Crl:CD(SD)BR (30/sex/group) provided DRM in the diet at concentrations of 0, 0.6, 6.0, and 30%. These dietary levels corresponded to overall average dosages of approximately 400, 3900, and 17,800 mg/kg/day for F0 males (premating) and 480, 4600, and 20,700 mg/kg/day for F0 females, respectively. Prior to mating, males and females of the F0 generation were treated for 10 and 2 weeks, respectively. Treatment of males continued throughout mating and until termination (approximately 3 weeks after mating). Treatment of the females was continued throughout gestation and through lactation day 21. The females were killed after raising their young to weaning at 21 days of age. Food consumption was measured weekly throughout the study (except during mating) and body weights were recorded at least weekly during premating, gestation, and lactation. Reproductive parameters including estrus cycle duration, mating performance, fertility, gestation length, parturition, and gestation index were evaluated. Litter size and offspring body weights were recorded, offspring viability indices were calculated, and physical development (vaginal opening and preputial separation) was assessed for the F1 generation. All adult F0 and F1 animals were subjected to a detailed necropsy. DRM treatment had no effect on estrus cycles or reproductive performance including mating performance, fertility, gestation length, parturition, or gestation index. Litter size, sex ratio, and offspring viability indices were similarly unaffected and there were no effects of DRM treatment on the physical development of F1 animals. PMID- 11407939 TI - Regulatory action criteria for filth and other extraneous materials v. strategy for evaluating hazardous and nonhazardous filth. AB - The U.S. Food and Drug Administration (FDA) uses regulatory action criteria for filth and extraneous materials to evaluate adulteration of food products. The criteria are organized into three categories: health hazards, indicators of insanitation, and natural or unavoidable defects. The health hazard category includes criteria for physical, chemical, and microbiological hazards associated with filth and extraneous materials. The health hazard category encompasses criteria for HACCP (Hazard Analysis and Critical Control Point) hazards and HACCP contributing factors. The indicators of insanitation category includes criteria for visibly objectionable contaminants, contamination from commensal pests, and other types of contamination that are associated with insanitary conditions in food processing and storage facilities. The natural or unavoidable category includes criteria for harmless, naturally occurring defects and contaminants. A decision tree is presented for the sequential application of regulatory action criteria for filth and extraneous materials associated with each category and with each type of filth or extraneous material in the three categories. This final report of a series in the development of a transparent science base for a revised FDA regulatory policy in the area of filth and extraneous materials in food includes a comprehensive list of the references that form the science base for the FDA regulatory policy. PMID- 11407940 TI - Early indicators of response in biologically based risk assessment for nongenotoxic carcinogens. AB - The proposed existence of dose-response thresholds for nongenotoxic carcinogens has led to a major controversy in the risk extrapolation process. To resolve this debate, there has been a significant investment in mechanism-based risk assessment research. The ability to utilize this mechanistic research for risk assessment procedures is still limited and may not warrant the expense. Alternatively, an approach can be used to identify dose-response thresholds through the utilization of sensitive indicators of biological response. This approach does not rely upon a mechanistic framework for the development of pathology, is solely dependent on already existing technology, and takes into account the possibility of background levels of pathway activation. For this approach, sensitive biochemical responses need to be identified and linked to the introduction of the toxicant through dose response, by time of response, and, when possible, through a proposed biochemical mechanism. The weakness of this approach is that more sensitive unidentified responses may exist requiring that a safety factor of 10 be used to define a NOEL. For dioxin-like compounds, using a surrogate marker of response CYP1A1 induction, this approach yields an estimate of the acceptable daily intake of 5-50 fg/kg/day. This limit is remarkably similar to the results of the original EPA linear extrapolation (6 fg/kg/day). A similar approach can be used for other nongenotoxic carcinogens and the analysis can be completed within 1 year. PMID- 11407941 TI - Ethylene "had no available [Salmonella] mutagenicity data". PMID- 11407942 TI - Introduction. The laboratory mouse in cancer research. PMID- 11407943 TI - Genetically engineered mouse models of astrocytoma: GEMs in the rough? AB - Astrocytomas are the most common form of brain cancer and are essentially incurable due to their diffusely infiltrative nature. Mouse models of astrocytoma provide a useful system for understanding tumorigenesis of astrocytomas and for designing and testing new therapies. Although molecular genetic alterations have been characterized in human astrocytomas, many of the mice engineered with these mutations do not develop astrocytomas. Recently, successful modeling of astrocytoma in the mouse has suggested that the combination of molecular alterations, the cell type in which the alterations take place, and the strain background all play a role in generating a model of astrocytoma. PMID- 11407944 TI - Modeling myeloid leukemia tumor suppressor gene inactivation in the mouse. AB - Introducing dominant oncogenic alterations uncovered in human myeloid malignancies into the mouse germline provides a powerful approach for studying leukemogenesis. However, little is known about how gene inactivation contributes to the development of myeloid malignancies. We describe how Nf1 mutant mice provide one example in which disrupting a tumor suppressor gene has been used to generate an informative murine leukemia model. We also discuss how chromosome engineering technologies are being harnessed to model the segmental deletions found in myeloid malignancies, and how these approaches can be combined with retrovirally medicated insertional mutagenesis to generate new models and for gene discovery. PMID- 11407945 TI - The genetics of pancreatic adenocarcinoma: a roadmap for a mouse model. AB - Pancreatic cancer is among the leading causes of cancer death. Although a genetic profile for pancreatic cancer is emerging, many biological aspects of this disease are poorly understood. Indeed, fundamental questions regarding progenitor cell lineages, host stromal milieu, and the role of specific genetic alterations in tumor progression remain unresolved. A mouse model engineered with signature mutations would provide a powerful ally in the study of pancreatic cancer biology and may guide improved prognostic assessment and treatment for the human disease. In this review, we discuss the molecular basis for normal pancreatic development and the genetics of human pancreatic adenocarcinoma in the hope of charting a course for the development of a faithful mouse model for this lethal cancer. PMID- 11407946 TI - Mouse models for human familial adenomatous polyposis. AB - Colorectal cancer (CRC) is the second most frequent type of cancer in the Western hemisphere. In the United States alone, it is estimated that 150 000 new cases are detected every year and more than 65 000 patients die from complications associated with this cancer. Identification of genes implicated in the initiation and progression of colorectal cancer in humans has prompted the generation of mouse models for this cancer. We will provide a brief overview of these mouse models for CRC and what they have contributed to our understanding of the events involved in the initiation and progression of this cancer. PMID- 11407947 TI - Modeling chromosomal instability and epithelial carcinogenesis in the telomerase deficient mouse. AB - Human carcinomas are intimately linked to advancing age. These cancers have complex cytogenetic profiles, including aneuploidy and chromosomal structural aberrations. While aged humans sustain a high rate of carcinomas, mice bearing common tumor suppressor gene mutations typically develop soft tissue sarcomas and lymphomas. One marked species distinction between human and mouse that bears on the predisposition to carcinogenesis lies in the radical differences in length and regulation of the telomere, nucleoprotein complexes that cap the ends of eukaryotic chromosomes. Recent cancer modeling studies in the telomerase knockout p53 mutant mice revealed that telomere dynamics might be relevant to carcinogenesis. In these mice, there is a shift in the tumor spectrum towards epithelial carcinomas, and these cancers emerge with complex cytogenetic profiles classical for human carcinomas. In this review, we suggest that the mechanism of fusion-bridge-breakage-translocation, triggered by critically short telomeres, may be one of the generators of genomic instability commonly seen in human carcinomas. PMID- 11407948 TI - The genetics and pathology of mouse mammary cancer. PMID- 11407949 TI - Autochthonous mouse models for prostate cancer: past, present and future. AB - Prostate cancer continues to be the second leading cancer related death among men. In order to more fully develop effective prevention and intervention strategies for this prevalent disease, the underlying molecular mechanisms of initiation, progression and metastatic spread must be understood. To this end mouse models have an essential role in prostate cancer research in that they can closely mimic the pathological and biochemical features of the disease. In this review we discuss the history of autochthonous murine models of prostate cancer, the essentials of the idealized mouse model for prostate cancer and speculate on approaches towards this goal. PMID- 11407950 TI - Genetic dissection of melanoma pathways in the mouse. AB - The frequent loss of the INK4a/ARF locus, encoding for both p16(INK4a)and p19(ARF)in human melanoma, raises the question as to which INK4a/ARF gene product functions to suppress melanoma-genesis in vivo. Studies in the mouse have shown that activated RAS mutation can cooperate with INK4a(Delta 2/3)deficiency (null for both p16(INK4a)and p19(ARF)) to promote development of melanoma, and these melanomas retain wild-type p53. Given the functional link between p19(ARF)and p53, we have now shown that activated RAS can also cooperate with p53 deficiency to produce melanoma in the mouse. Moreover, genome-wide analysis of RAS-induced p53 mutant melanomas reveals alterations of key components governing RB-regulated G1/S transition, such as c-Myc. These experimental findings suggest that both RB and p53 pathways function to suppress melanocyte transformation in vivo in the mouse. PMID- 11407951 TI - The quality of life of older people with epilepsy: findings from a UK community study. AB - The impact of epilepsy on the quality of life of older people with epilepsy has rarely been investigated. As part of a large prevalence study of epilepsy conducted in one UK Health Region, we investigated the burden of their condition in older compared with younger people. A second analysis compared quality of life in those men and women diagnosed after the age of retirement from the workforce compared with those diagnosed before that age. Data were collected from men and women with epilepsy and from their primary and secondary care physicians. There were few differences between older and younger people with regard to their reported quality of life, though younger people were more likely to report feeling stigmatized by their condition. Older people with epilepsy diagnosed in later life were more anxious and depressed than those diagnosed earlier and their overall perception of quality of life was more likely to be negative. Our data emphasize that older people do not necessarily experience poorer quality of life than younger people, but those first diagnosed in later life do appear to have a quality of life which is more impaired. Consideration should be paid to the important psychosocial consequences of epilepsy in the older person. PMID- 11407952 TI - Quality of life of patients with epilepsy (Hungarian survey). AB - We assessed the quality of life (QOL) of patients with epilepsy using the Quality of Life in Epilepsy Inventory (QOLIE-31). As the first step we compared our results with the data from an American survey in order to validate the test in Hungary. The results show that the Hungarian values were lower but that they followed the same trends as the American data. There was only one controversial result in the question-group of the 'the effects of treatment', which could be explained by the differences in habits and conventions, opportunities and expectations between Hungarian and American epileptic patients. We found significant differences in many aspects of quality of life with respect to (a) gender (general quality of life, seizure worry), (b) pharmacological treatment form (cognitive functions, medication effects, total score and social and role functioning) and (c) economic activity of patients (cognitive functions, emotional well-being, energy/fatigue, medication effects, overall quality of life, overall scores, seizure worry, social and role functioning). We have tried to explain the differences found by taking either the characteristics of epilepsy or the social background of the epileptic patient into consideration. Based on previous knowledge we have tried to define the situations where the assessment of quality of life for people with epilepsy, may be beneficial to their core. PMID- 11407953 TI - A randomized open-label study of gabapentin and lamotrigine in adults with learning disability and resistant epilepsy. AB - The aim of this study was to evaluate the efficacy and safety of gabapentin in patients with learning disabilities and resistant epilepsy, comparing the effects of gabapentin with lamotrigine on efficacy, behaviour and mood. An open-label, randomized, parallel group, multicentre add-on study comparing gabapentin with lamotrigine in 109 patients with drug-resistant localization-related epilepsy and learning disabilities was conducted: 39 patients were randomized to gabapentin and 44 to lamotrigine. The study population had a range of learning disabilities and severe partial epilepsy. The percentage of patients achieving a greater than or equal to 50% reduction in seizure frequency on gabapentin was 50%, (mean reduction in seizures was 51%). Compared to 48.6% of lamotrigine patients, no statistically significant treatment differences could be identified. The safety profile of both drugs was consistent with that seen in previous clinical trials. Carer-rated visual analogue scales detected significant improvements (P< 0.05) for the gabapentin-treated patients in seizure severity, attention, general health and sleeping pattern, while for lamotrigine seizure severity improved significantly. For learning disabled patients with resistant epilepsy, gabapentin and lamotrigine provide safe and effective treatment, with positive benefits on behaviour. PMID- 11407954 TI - Status epilepticus arising de novo in hospitalized patients: an analysis of 41 patients. AB - Most of the information on predisposing factors and mortality in status epilepticus (SE) arises from data obtained from patients presenting to the casualty department. However, another population which is frequently seen by consultative neurologists are medically ill patients who develop SE while in hospital. These patients are often notoriously difficult to treat once SE arises. We sought to characterize patients at risk for SE arising when they are hospitalized for other reasons. By doing this, risk factors for developing SE and prognostic indicators might be determined. We retrospectively reviewed records from three urban hospitals in the United States to identify hospitalized patients developing SE over a 1 year period. SE was defined as a clinical seizure lasting 30 minutes or longer, or repeated seizures without recovery. Patients who were admitted in SE or for an epilepsy-related problem, or who were less than 1 year old were excluded from the study. Forty-one patients with in-hospital SE were identified. There were 28 males and 13 females with an age range from 1 to 91 years (mean: 60 years, median: 65 years). The mean interval from hospital admission to the onset of status epilepticus was 26 days. Nineteen (46%) patients had a prior history of either epilepsy or symptomatic seizures, and of these, 10 were inadequately treated as judged by serum anticonvulsant levels at the time SE developed. Focal brain abnormality was present in 26 (63%) patients, the most common of which was stroke (17 patients ). Major metabolic derangements including hypoxia, electrolyte imbalance, hepatic encephalopathy, and sepsis were present in 23 (56%) patients. Eleven (27%) patients were being treated with theophylline preparations at the time SE developed. Mortality in this group of patients with in-hospital SE was 61% (25 deaths), with about one-third dying while in status, and two-thirds dying subsequently in hospital. In this retrospective study, there was no clear relationship between mortality and the duration of SE in this group of patients. In-hospital development of SE is usually related to underlying focal brain abnormality, especially stroke, in combination with systemic metabolic derangement. Prognosis is poor, and appears to be more related to underlying conditions rather than to status duration. More accurate prospective studies are warranted. PMID- 11407955 TI - Effective and safe but forgotten: methsuximide in intractable epilepsies in childhood. AB - The efficacy and safety of methsuximide (MSM) was evaluated in children with intractable epilepsies in a prospective uncontrolled study. MSM was added to the therapeutic regimen of 112 children with intractable epilepsy under inpatient conditions, all of whom were therapeutically refractory to various first-line antiepileptic drugs (AED) or combinations of other AED. Titration of MSM was performed following a uniform protocol. Administration of MSM resulted in a 50% or greater reduction in seizure frequency in 40 patients after a short-term observation period (mean 9.1 weeks). After a mean of 3.7 years, the rate of seizures and side effects were re-evaluated in 39 patients who were still receiving MSM as part of their antiepileptic regimen. Twenty two of these patients derived long-term benefit from MSM. In patients with good seizure control, fasting plasma levels of N-desmethylmethsuximide, the principal active metabolite of MSM, were 25.3-44.7 mg l(-1)(mean 36.0 mg l(-1)). Thus effective plasma levels of N-desmethylmethsuximide in children were found to be higher than previously described. Forty one of 112 patients (28.9%) developed side effects during MSM treatment. No serious or irreversible side effects were seen. Our study demonstrates the value of MSM as an 'add-on' drug in intractable epilepsies. PMID- 11407956 TI - Cognitive dysfunction of right hemisphere-like Todd's paralysis after status epilepticus: a case report. AB - We describe a case with symptoms of transient diffuse right hemisphere dysfunction (hemispatial neglect, dyscalculia, and disturbance of both spatial construction and visuospatial perception) occurring after status epilepticus. The clinical picture of this case suggested to us that these features could be understood as a variant of Todd's paralysis. PMID- 11407957 TI - Ictal deafness--a report of three cases. AB - Though negative phenomena like motor inhibition, blindness or aphasia are described as an ictal manifestation of seizures, ictal deafness has not been reported so far. We observed transient ictal deafness in three cases of partial seizures. One of them had seizure spread to the temporal lobe to produce a complex partial seizure. Two of them have CT-detected lesions in the left temporal/parietal area. The other one had left temporal focus on EEG with a normal imaging study. The mechanism of such ictal negative phenomena is unclear. An epileptic focus around the primary auditory cortex, dampening its receptive ability may manifest as cortical deafness. PMID- 11407958 TI - Orbito-frontal epilepsy masquerading as temporal lobe epilepsy-a case report. AB - Temporal lobectomy fails to control seizures in a considerable percentage of patients who do not have hippocampal sclerosis. One theoretical reason for failure of surgery is that some of these patients may in fact have extratemporal epilepsy. We present a 28-year-old woman with clinical and scalp electroencephalogram (EEG) evidence of right temporal lobe epilepsy (TLE) supported by functional imaging with interictal positron emission tomography (PET) and ictal single-photon emission computerized tomography (SPECT). An invasive EEG monitoring was prompted by the discovery of a small right orbito frontal lesion on MRI. Monitoring documented seizure onset at the lesion, with rapid right temporal involvement. The patient was almost seizure-free after a lesionectomy. The index of suspicion of orbito-frontal epilepsy should be high in patients with apparent TLE when the scalp EEG and neuroimaging data are not congruent, or if temporal lobe pathology cannot be identified on structural imaging. PMID- 11407959 TI - CPD - education and self-assessment: functional imaging in epilepsy. AB - Functional imaging plays a growing role in the clinical assessment and research investigation of patients with epilepsy. This article reviews the literature on functional MRI (fMRI) investigation of EEG activity, fMRI evaluation of cognitive and motor functions, magnetic resonance spectroscopy (MRS), single photon emission computed tomography (SPECT) and positron emission tomography (PET) in epilepsy. The place of these techniques in clinical evaluation and their contribution to a better neurobiological understanding of epilepsy are discussed. PMID- 11407960 TI - Case reports--functional imaging in epilepsy. PMID- 11407962 TI - The effects of selected classical music on self-disclosure. AB - A cognitive theory of emotion and aesthetics in music suggests that listening to music is cognitively and in turn emotionally arousing. In addition a theory of inhibition and psychosomatic disease suggests that disclosing personal and traumatic information is psychologically and physically beneficial. This study examined the effect of music within a disclosure setting on the use of cognitive and emotional language in 85 undergraduates. Participants in both the background music and no music conditions wrote or spoke into a tape recorder, about the most significant event or experience of their lives. In addition to language, self reported moods, subject impressions about the disclosure experience, and the environment were examined. Results indicate that background music had an effect on the disclosure topics chosen, promoted cognitive suggestion and expression, and increased the enjoyment of listening to classical music. PMID- 11407963 TI - Gender differences among newborns on a transient otoacoustic emissions test for hearing. AB - The purpose of this study was to examine gender differences with regard to cochlea sensitivity as measured by the transient evoked otoacoustic emissions hearing screening procedure. During this test, a sudden burst of sound is presented at between 78 and 83 dB SPL which stimulates the entire basilar membrane. This in turn excites the outer hair cells in the cochlea and causes an echo-type response which is recorded by a microphone in a probe placed in the ear canal. This test is used to screen for peripheral hearing loss. Subjects ( N = 350) for this project were healthy, full-term newborns (38-42 weeks gestation) in the first 48 hours of life who had bilaterally passed the transient evoked otoacoustic emissions (TEOAE) screening test. Male (n = 170) and female (n = 180) infants were selected randomly from all babies born during a 3-month period who met the criteria at a large birthing hospital. Responses to TEOAE stimuli were recorded at 1.6 kHz, 2.4 kHz, 3.2 kHz, and 4.0 kHz. The responses were recorded as decibel levels indicating a signal-to-noise ratio. These decibel levels were used in a three-way ANOVA with repeated measures comparing gender, ear, and frequency level. Results indicated significant differences due to gender (female hearing more sensitive than male) and frequency (least sensitive hearing recorded at 1.6 kHz, most sensitive hearing recorded at 3.2 kHz). A significant two-way interaction indicated that differences in hearing sensitivity between genders increased as the frequency increased. PMID- 11407964 TI - The effect of presentation and accompaniment styles on attentional and responsive behaviors of participantswith dementia diagnoses. AB - Eight persons (7 females & 1 male) with moderately-severe to severe dementia diagnoses residing at an urban healthcare facility, participated in this study. Informed consent procedures were followed and agreed upon by participants and their families. The participants ranged in age from 73-90 years, with an average age of 81 years. An experienced board-certified music therapist led 16 singalong sessions. The sessions were divided into 4 sessions of each of the following 4 conditions in randomized order of presentation: (a) live music of simple guitar accompaniment, (b) live music of complex guitar accompaniment, (c) recorded music of simple guitar accompaniment, and (d) recorded music of complex guitar accompaniment. Results indicated no significant differences in the amount of singing by the group for the 4 conditions. However, there were significant differences in group behavior related to the 4 conditions before and after song presentations, including attention, leaving the group, reading lyrics, compliments, and applause. Implications for music therapy training and programming are discussed. PMID- 11407965 TI - The effect of the bonny method of guided imagery and music on the mood and life quality of cancer patients. AB - Cancer patients continue to experience elevated levels of emotional distress, even after cancer treatment. Elevated emotional distress negatively impacts immune and endocrine functions and decreases life quality. This study explored the effectiveness of GIM in alleviating mood disturbance and improving quality of life in cancer patients. Eight volunteers with a cancer history were randomly assigned to either an experimental or a wait-list control group. Experimental subjects individually participated in 10 weekly GIM sessions. All subjects completed the Profile of Mood States (POMS) and Quality of Life-Cancer (QOL-CA) questionnaires pretest, posttest, and at a 6-week follow-up. Individuals who participated in GIM sessions scored better on both mood scores and quality of life scores at posttest than those participating in the control group. Additionally, mood and quality of life scores continued to improve in the experimental group, even after sessions were complete. Results indicate that GIM was effective in improving mood and quality of life in these cancer patients. PMID- 11407966 TI - Music therapy in the age of enlightenment. AB - As music therapists continue to discover more about the therapeutic powers of music, it is interesting now and then to look to the past in order to seek the roots of our contemporary practices. In this regard, the writings of eighteenth century physicians are pivotal in the development of music therapy, for it was these individuals who first began to depend greatly upon scientific experimentation and observation to formulate their procedures. Representative of this stage in the history of music therapy are the findings of the renowned London physician Richard Brocklesby, the only doctor to write a treatise on music therapy in eighteenth-century England. The subjects treated by Brocklesby in his Reflections on the Power of Music (1749) include his musical remedies for the excesses of various emotions-particularly fear, excessive joy, and excessive sadness. He also discusses his musical remedies for diseases of the mind recognized in the eighteenth century-delirium, frenzy, melancholia, and maniacal cases. He considers music as well an aid to the elderly and to pregnant women. In short, Brocklesby provides a lively account of the curative powers of music as viewed in the mid-eighteenth century by an excellent medical mind. PMID- 11407968 TI - Selective pick-up of increased iron by deferoxamine-coupled cellulose abrogates the iron-driven induction of matrix-degrading metalloproteinase 1 and lipid peroxidation in human dermal fibroblasts in vitro: a new dressing concept. AB - Using atomic absorption spectrum analysis, we found iron levels in exudates from chronic wounds to be significantly increased (3.71 +/- 1.56 micromol per g protein) compared to wound fluids from acute wounds derived from blister fluids (1.15 +/- 0.62 micromol per g protein, p < 0.02), drainage fluids of acute wounds (0.87 +/- 0.34 micromol per g protein, p < 0.002), and pooled human plasma of 50 volunteers (0.42 micromol per g protein). Increased free iron and an increase in reactive oxygen species released from neutrophils represent pathogenic key steps that --via the Fenton reaction - are thought to be responsible for the persistent inflammation, increased connective tissue degradation, and lipid peroxidation contributing to the prooxidant hostile microenvironment of chronic venous leg ulcers. We herein designed a selective pick-up dressing for iron ions by covalently binding deferoxamine to cellulose. No leakage occurred following gamma sterilization of the dressing and, more importantly, the deferoxamine-coupled cellulose dressing retained its iron complexing properties sufficient to reduce iron levels found in chronic venous ulcers to levels comparable to those found in acute wounds. In order to study the functionality of the dressing, human dermal fibroblasts were exposed to a Fenton reaction mimicking combination of 220 microM Fe(III) citrate and 1 mM ascorbate resulting in a 4-fold induction of matrix degrading metalloproteinase 1 as determined by a matrix-degrading metalloproteinase 1 specific enzyme-linked immunosorbent assay. This induction was completely suppressed by dissolved deferoxamine at a concentration of 220 microM or by an equimolar amount of deferoxamine immobilized to cellulose. In addition, the Fe(III) citrate and ascorbate driven Fenton reaction resulted in an 8-fold increase in malondialdehyde, the major product of lipid peroxidation, as determined by high pressure liquid chromatography. This increase in malondialdehyde levels could be significantly reduced in the presence of the selective pick-up dressing coupled with deferoxamine suggesting that the deferoxamine dressing, in fact, prevents the development of a damaging prooxidant microenvironment and also protects from unfavorable consequences like matrix degrading metalloproteinase 1 and lipid peroxide induction. PMID- 11407969 TI - Aging and effects of ultraviolet A exposure may be quantified by fluorescence excitation spectroscopy in vivo. AB - The fluorescence properties of skin chromophores such as tryptophan and collagen cross-links might be useful markers of aging and photoaging. As the fluorescence of pepsin-digestible collagen cross-links was found to increase with aging and decrease with photoaging we investigated the characteristics of this dependence. In vivo fluorescence excitation spectra (emission at 380 nm) of SKH hairless mouse model skin are characterized by two bands centered near 295 nm and 335 nm due, respectively, to epidermal tryptophan moieties and pepsin-digestible collagen cross-links. Several groups of hairless mice were followed over a period of 18 mo to document changes in skin fluorescence with aging. Other groups of animals were exposed to either broad band or narrowband ultraviolet A radiation to determine the effects of ultraviolet A exposure on the fluorescence of the dermal collagen cross-links and to determine an action spectrum for the induced changes. We also found that the intensity of pepsin-digestible collagen cross links in vivo increases linearly with age and that the fluorescence of epidermal tryptophan decreases linearly with age. We found that the fluorescence of pepsin digestible collagen cross-links decreases immediately following exposure to ultraviolet A whereas epidermal tryptophan fluorescence increases. Both changes were dose dependent but the increase in tryptophan fluorescence occurred exclusively in young animals (2--6 mo old). We found that the ultraviolet-induced fluorescence decrease of pepsin-digestible collagen cross-links is wavelength specific. The action spectrum for the ultraviolet A effect on the in vivo fluorescence of pepsin-digestible collagen cross-links shows a distinct maximum at 335 nm that corresponds to the maximum in the fluorescence excitation spectrum due to pepsin-digestible collagen cross-links. Our results seem to indicate that in vivo fluorescence of epidermal tryptophan moieties and collagen cross-links in the dermal matrix may serve as markers for skin aging, for photoaging, and for immediate assessment of exposure to ultraviolet A radiation. PMID- 11407970 TI - Topographic variations in normal skin, as viewed by in vivo reflectance confocal microscopy. AB - Near-infrared confocal microscopy is a new tool that provides skin images in vivo, with high resolution and contrast at a specific depth. Regional variations in live human skin viewed by confocal microscope have not been studied so far. In vivo reflectance confocal microscopy was performed in 10 adults (eight males, two females) of various skin phototypes. Six topographic sites were studied in each subject: forehead, cheek, inner and outer forearm surfaces, lower back and leg. Epidermal thickness at suprapapillary epidermal plates and rete pegs was measured during real-time imaging and the number and diameter of epidermal keratinocytes in each epidermal cell layer as well as the characteristics of dermal papillae were defined from the grabbed images. Stratum corneum appeared brighter in sun exposed than in sun-protected areas and particularly pronounced in heavily pigmented individuals. The epidermal thickness at rete pegs, but not the suprapapillary epidermal plate, was greater in sun-exposed areas than in sun protected sites except forearm flexor surface. The en face numerical density of granular keratinocytes is greater on the face as compared with all other sites, whereas the surface density of spinous keratinocytes is greater on sun-protected sites. Additionally, the number of basal keratinocytes per millimeter length of dermoepidermal junction is greater in sun exposed areas. Interestingly, the dermal papillae shape varies and their sizes increase in circumference from sun exposed to sun-protected sites, as observed at a specific depth below the stratum corneum. In summary, our results demonstrate that near infra-red reflectance confocal microscopy is a feasible tool for microscopic analysis of skin morphometry in vivo. PMID- 11407971 TI - Topically applied vitamin C enhances the mRNA level of collagens I and III, their processing enzymes and tissue inhibitor of matrix metalloproteinase 1 in the human dermis. AB - Ascorbic acid (vitamin C) is a cofactor required for the function of several hydroxylases and monooxygenases. It is not synthesized in humans and some other animal species and has to be provided by diet or pharmacologic means. Its absence is responsible for scurvy, a condition related in its initial phases to a defective synthesis of collagen by the reduced function of prolylhydroxylase and production of collagen polypeptides lacking hydroxyproline, therefore, they are unable to assemble into stable triple-helical collagen molecules. In fibroblast cultures, vitamin C also stimulates collagen production by increasing the steady state level of mRNA of collagen types I and III through enhanced transcription and prolonged half-life of the transcripts. The aim of the experimental work has been to evaluate the effect on dermal cells of a preparation of vitamin C topically applied on one side vs placebo on the other side of the dorsal face of the upper forearm of postmenopausal women. Biopsies were collected on both sides and the level of mRNA measured by non competitive reverse transcription polymerase chain reaction made quantitative by the simultaneous transcription and amplification of synthetic RNA used as internal standards. The mRNA of collagen type I and type III were increased to a similar extent by vitamin C and that of three post-translational enzymes, the carboxy- and amino-procollagen proteinases and lysyloxidase similarly increased. The mRNA of decorin was also stimulated, but elastin, and fibrillin 1 and 2 were not modified by the vitamin. The expression of matrix metalloproteinases 1, 2, and 9 was not significantly changed, but an increased level of tissue inhibitor of matrix metalloproteinase 1 mRNA was observed without modification of tissue inhibitor of matrix metalloproteinase 2 mRNA. The stimulating activity of topical vitamin C was most conspicuous in the women with the lowest dietary intake of the vitamin and unrelated to the level of actinic damage. The results indicate that the functional activity of the dermal cells is not maximal in postmenopausal women and can be increased. PMID- 11407972 TI - Keratinocyte apoptosis induced by ultraviolet B radiation and CD95 ligation -- differential protection through epidermal growth factor receptor activation and Bcl-x(L) expression. AB - Previous work has shown that activation of the epidermal growth factor receptor by endogenous or exogenous signals markedly enhances survival of cultured keratinocytes upon cellular stress such as passaging. This is due, in part, to epidermal-growth-factor-receptor-dependent expression of Bcl-x(L), an antiapoptotic Bcl-2 homolog. In this study we tested whether epidermal-growth factor-receptor-dependent signal transduction and attendant Bcl-x(L) expression affected survival of human keratinocytes upon exposure to a frequently encountered apoptotic stimulus, radiation with ultraviolet B. We describe that blocking epidermal-growth-factor-receptor-dependent signal transduction sensitized normal keratinocytes to undergo apoptosis upon ultraviolet B radiation with solar light characteristics. Forced expression of Bcl-x(L) partially but significantly inhibited ultraviolet-B-induced apoptosis of immortalized keratinocytes (HaCaT). Bcl-x(L) overexpression afforded no protection to HaCaT cells against apoptosis induced by binding of an agonist antibody to the death receptor CD95, however. CD95 activation has previously been shown to functionally contribute to apoptosis in ultraviolet-irradiated keratinocytes. These results indicate that epidermal growth factor receptor activation and attendant Bcl-x(L) expression provided a physiologically relevant protective pathway of keratinocytes against ultraviolet-induced but not CD95-dependent apoptosis. PMID- 11407973 TI - Possible regulation of telomerase activity by transcription and alternative splicing of telomerase reverse transcriptase in human melanoma. AB - To investigate the regulatory mechanisms of telomerase activity in human melanoma cells, we assessed the enzyme's catalytic activity and the expression of the telomerase subunits, the human telomerase RNA, the human telomerase-associated protein, and the human telomerase reverse transcriptase, in 52 melanoma lesions. Eight normal skin specimens were also studied. Telomerase activity was detected in 84.6% of melanomas, whereas all skin specimens were telomerase negative. Human telomerase-associated protein mRNA and human telomerase RNA were constitutively expressed in all melanoma and skin specimens. Although at a variable level of expression, human telomerase reverse transcriptase mRNA was detected in all but one melanomas, whereas it was never present in skin samples. Reverse transcriptase-polymerase chain reaction experiments were performed using primers within the reverse transcriptase domain of human telomerase reverse transcriptase and revealed the presence of multiple alternatively spliced transcripts in melanoma specimens. Among the 44 telomerase-positive melanomas, one showed the full-length transcript alone whereas in all other specimens a full-length message was present with different combinations of alternatively spliced variants. In these tumors the expression of the full-length transcript was generally equal to or higher than that of the alternatively spliced variants. The ratio full-length transcript to alternatively spliced species ranged from 0.6 to 5.26, with a median value of 1.18. Among the seven telomerase-negative melanomas, one displayed the beta deletion transcript alone, whereas in the remaining six tumors weak expression of the full-length transcript and a more abundant level of alternatively spliced transcripts were found. In these cases human telomerase reverse transcriptase ratio ranged from 0.09 to 1.1, with a median value of 0.40. The results suggest that transcription and alternative splicing of human telomerase reverse transcriptase are regulatory mechanisms controlling telomerase activity in melanoma. PMID- 11407974 TI - The interferon inhibiting cytokine IK is overexpressed in cutaneous T cell lymphoma derived tumor cells that fail to upregulate major histocompatibility complex class II upon interferon-gamma stimulation. AB - Cutaneous T cell lymphomas are characterized by an accumulation of malignant clonal lymphocytes in the skin and occasionally in the blood. We compared gene transcription profiles from cultured clonal lymphocytes with autologous healthy blood lymphocytes by microarray hybridization. Cutaneous T cell lymphoma derived cells transcribed high amounts of an interferon inhibiting cytokine factor. The presence of an interferon inhibiting cytokine factor was confirmed in 12 skin biopsies of mycosis fungoides and Sezary syndrome derived blood lymphocytes by reverse transcriptase-polymerase chain reaction. The presence of interferon inhibiting cytokine factor mRNA in Sezary syndrome derived lymphocytes was associated with a lack of HLA class II upregulation after stimulation with interferon-alpha and interferon-gamma. This was not due to a loss of the interferon signaling cascade as the presence of interferon-signaling components was confirmed by reverse transcriptase--polymerase chain reaction on the transcriptional level. The elevated constitutive interferon inhibiting cytokine factor expression observed in cutaneous T cell lymphoma derived cells was insensitive to interferon-gamma stimulation, but was enhanced in normal peripheral blood mononuclear cells. We suggest that interferon inhibiting cytokine factor contributes to the lack of HLA class II upregulation in lymphoma cells. Interferon inhibiting cytokine factor may participate in providing a microenvironment at the tumor site insensitive to interferon-gamma stimulation and thus prevents an efficient local immune response. PMID- 11407975 TI - Activation of murine epidermal V gamma 5/V delta 1-TCR(+) T cell lines by Glu-Tyr polypeptides. AB - The physiologic role of gamma delta-T-cell-receptor-bearing cells and the T cell receptor ligands that they recognize is still poorly understood. Previous studies have suggested that one possible antigen for gamma delta-TCR(+) cells is the random copolymer poly-glutamic acid-tyrosine (poly-Glu-Tyr), because poly-Glu-Tyr reactive gamma delta-TCR(+) hybridoma cells were produced from poly-Glu-Tyr immunized mice. We have found, however, that clonal V gamma 5/V delta 1-TCR(+) epidermal T cell lines from nonimmune mice also respond to poly-Glu-Tyr by producing cytokines. Other amino acid homopolymers, copolymers, and tripolymers were not stimulatory for the V gamma 5/V delta 1-TCR(+) epidermal T cells, except for poly-glutamic acid-alanine-tyrosine (poly-Glu-Ala-Tyr). Of the poly-Glu-Tyr and poly-Glu-Ala-Tyr polymers, only those that contained Glu and Tyr in an equimolar ratio were stimulatory. The cytokine interleukin-2 was strictly required for the responses to poly-Glu-Ala-Tyr, whereas the responses to poly-Glu Tyr were merely enhanced with interleukin-2. The response to poly-Glu-Tyr was also enhanced by crosslinking CD28 molecules with plate-bound anti-CD28 crosslinking antibody. This finding suggests that the poly-Glu-Tyr response has a partial dependence on CD28-mediated costimulation, a characteristic of TCR dependent responses. Consistent with this observation, V gamma 5/V delta 1-TCR loss variants of the epidermal T cell line could no longer respond to poly-Glu Tyr. The unpredicted responses of epidermal gamma delta-TCR(+) T cells to poly Glu-Tyr and poly-Glu-Ala-Tyr demonstrate that the functions of these cells potentially can be triggered by peptidic ligands, probably through a TCR-mediated process. PMID- 11407976 TI - Patients with allergic and irritant contact dermatitis are characterized by striking change of iron and oxidized glutathione status in nonlesional area of the skin. AB - To assess the consequences of oxidative stress in allergic and irritant contact dermatitis, we compared the iron level, unsaturated iron-binding capacity, total iron binding capacity, the percentage saturation of iron-binding capacity, the amount of diene conjugates as well as the amounts of total glutathione, reduced glutathione, oxidized glutathione, and the oxidized glutathione/reduced glutathione ratio in skin homogenate from lesional and nonlesional skin. Lesional skin samples were obtained from positive patch test sites to 5% NiSO4 in five subjects, and from chronic contact dermatitis lesions on the hands, which had exacerbated over 3--9 wk in six subjects. Contact dermatitis caused at least a 4 fold increase in the iron level in the lesional skin area compared with the nonlesional skin area (p < 0.02). The increase in the iron level depended on the duration of contact dermatitis and was accompanied by high unsaturated iron binding capacity and total iron-binding capacity values in the positive patch test sites (p < 0.05), and by a high percentage saturation value in the chronic contact dermatitis lesions (p < 0.05). We found high indices for iron, total iron binding capacity and diene conjugates in the apparently healthy skin of the patients with persistent contact dermatitis that significantly (p < 0.05) exceeded the corresponding values in the patients with only patch test reactions. In summary, we have succeeded in providing evidence that generalized oxidative damage of the skin occurs as a consequence of contact dermatitis in a restricted area. PMID- 11407977 TI - HLA-A2 restricted, melanocyte-specific CD8(+) T lymphocytes detected in vitiligo patients are related to disease activity and are predominantly directed against MelanA/MART1. AB - Vitiligo is a skin and hair disorder characterized by circumscribed depigmented lesions due to lack of melanocytes in the respective areas. It has been suggested that vitiligo is caused by an autoimmune-mediated destruction of melanocytes. Recently, the presence of a high frequency of skin-homing melanocyte-specific cytotoxic T lymphocytes in the peripheral blood of patients with vitiligo was reported. Our study examines the frequency of melanocyte-specific cytotoxic T lymphocytes in vitiligo patients and its relationship to disease activity. Thirty two patients with moderate to active vitiligo and 17 control subjects were included. Melanocyte specific reactive CD8(+) T cells were identified by enzyme linked immunospot assay after stimulation with five peptides from gp100, four peptides from MelanA/MART1, and two peptides from tyrosinase. In selected patients, intracellular interferon-gamma staining for the detection of specific reactive CD8(+) T cells was additionally performed. In seven of 10 patients (70%) with actively progressive disease CD8(+) T cells directed against melanocyte epitopes were detected, whereas only in four of 22 patients (18%) with moderate disease activity such specific reactivity was found. MelanA/MART1 peptides were immunodominant in nine patients reacting against EAAGIGILTV and three patients reacting against ILTVILGVL. Intracellular interferon-gamma staining confirmed the findings obtained by the enzyme-linked immunospot technique. The present study supports the hypothesis that vitiligo is a cytotoxic T lymphocyte-mediated autoimmune disease. The presence of melanocyte-specific reactive CD8(+) T cells seems to be closely related to disease activity. PMID- 11407978 TI - alpha v beta 6 Integrin upregulates matrix metalloproteinase 9 and promotes migration of normal oral keratinocytes. AB - The integrin alpha v beta 6 is a fibronectin receptor that is undetectable on normal keratinocytes in situ, but is increased significantly in wound healing and in culture-established keratinocytes, suggesting that it may promote changes associated with cell motility. Using normal human oral keratinocytes we have shown that cultured cells express relatively high levels of alpha v beta 6 and this integrin has a functional role in both cell adhesion and migration towards fibronectin. We provide experimental evidence that the increased expression of alpha v beta 6 by normal human oral keratinocytes results in coordinate changes, which promote a more migratory phenotype. Thus increased expression of alpha v beta 6 results in a fibronectin-dependent increase in pro-matrix metalloproteinase 9, matrix metalloproteinase 9 activity increases normal human oral keratinocyte migration, and this may be further dependent on plasmin activation. The results suggest a key role for alpha v beta 6 in these processes and indicate a coordinated link between alpha v beta 6 expression and upregulation of matrix metalloproteinase 9. It appears that alpha v beta 6 may function in normal human oral keratinocyte migration through matrix metalloproteinase-9-dependent and -independent mechanisms. PMID- 11407979 TI - Calmodulin-like skin protein: a new marker of keratinocyte differentiation. AB - The expression of the calmodulin-like skin protein, a recently discovered new skin-specific calcium binding protein, was studied in cultured keratinocytes, reconstructed human epidermis, and normal human skin. Using a calmodulin-like skin protein specific polyclonal antibody and Western blot analysis we could show that in cultured keratinocytes calmodulin-like skin protein expression is strongly induced after stimulating cell differentiation by increasing the medium calcium concentration. Known modulators of epidermal differentiation such as sodium butyrate and the synthetic retinoid CD 367 strongly affected calmodulin like skin protein expression. A more than 10-fold increase was observed in the presence of sodium butyrate, whereas CD 367 abolished almost completely calmodulin-like skin protein expression already at nanomolar concentrations. Calmodulin, another calcium binding protein that is expressed throughout the living layers of the epidermis, is not affected by these modulators. In normal human skin, calmodulin-like skin protein expression is restricted to the stratum granulosum and the lower layers of the stratum corneum. From these results we conclude that calmodulin-like skin protein is a new marker of late keratinocyte differentiation with a role distinct from calmodulin. PMID- 11407980 TI - Enhancing 1 alpha-hydroxylase activity with the 25-hydroxyvitamin D-1 alpha hydroxylase gene in cultured human keratinocytes and mouse skin. AB - 1 alpha,25-Dihydroxyvitamin D(3) (1 alpha,25(OH(2))D(3)) and its analogs are used to treat psoriasis because of their potent antiproliferative activity. They have the potential for causing hypercalcemia, however, and patients often become resistant to the drug. We examined the feasibility of enhancing the cutaneous production of 1 alpha,25(OH(2))D(3) using a human 25-hydroxyvitamin D-1 alpha hydroxylase (1 alpha-OHase) plasmid. The 1 alpha-OHase gene was fused to the green fluorescent protein gene (1 alpha-OHase-GFP) driven by the cytomegalovirus promoter. Transfection of cultured normal human keratinocytes with the 1 alpha OHase-GFP plasmid resulted in a marked increase in the expression of 1 alpha OHase-GFP in the mitochondria. Transfection of keratinocytes with 1 alpha-OHase GFP or 1 alpha-OHase plasmids in vitro enhanced the 1 alpha-OHase activity substantially and increased the sensitivity of the keratinocytes to the antiproliferative effect of 25(OH)D(3). The 1 alpha-OHase-GFP plasmid was topically applied to shaved C57/BL6 mice. Twenty-four hours after topical application, immunohistochemical analysis of the skin for 1 alpha-OHase-GFP revealed the presence of 1 alpha-OHase-GFP in the epidermis and epidermal appendages including the hair follicles. The results from this study offer a unique new approach for the topical treatment of hyperproliferative disorders such as psoriasis and skin cancer using the 1 alpha-OHase gene that could locally increase the production of 1 alpha,25(OH(2))D(3) without causing hypercalcemia or resistance. PMID- 11407981 TI - Ultraviolet radiation increases tropoelastin mRNA expression in the epidermis of human skin in vivo. AB - Photoaged skin contains elastotic materials in the upper reticular dermis. This phenomenon is commonly known as solar elastosis. Little is known about the mechanisms leading to the accumulation of elastotic materials in photoaged skin, however. In this study, it was demonstrated that ultraviolet irradiation induced tropoelastin mRNA expression in the keratinocytes of human skin in vivo and also in cultured human keratinocytes by in situ hybridization and reverse transcriptase polymerase chain reaction. It was also shown by northern blot analysis (n = 5) that there were increased tropoelastin mRNA levels in the forearm (sun-exposed) skin of elderly persons, compared with upper-inner arm (sun protected) skin of the same individuals. As demonstrated by in situ hybridization compared to sun-protected skin (upper-inner arm) (n = 5), tropoelastin mRNA expression in photoaged skin was higher in keratinocytes as well as in fibroblasts. Therefore, our results suggest that keratinocytes are another source of tropoelastin production after acute and chronic ultraviolet irradiation in human skin in vivo. PMID- 11407982 TI - Kit(+) melanocytes seem to contribute to melanocyte proliferation after UV exposure as precursor cells. AB - Melanogenesis caused by UV exposure is characterized by proliferation and differentiation of functioning melanocytes in epidermis. Recently, the existence of precursor melanocytes in normal skin was proposed immunohistochemically. Thus, this precursor melanocyte seems to proliferate and differentiate into mature pigmented melanocytes after UV exposure. To elucidate how these processes occur, we examined C57BL mouse epidermal sheets after UV exposure immunohistochemically. In normal epidermis, TRP-1(+) cells and Kit(+) cells were easily identified and the cellularities were 41.0 and 31.7 cells per mm(2), respectively. Only a few Mitf(+) cells and no TRP-2(+) cells were observed. On the first day after UV exposure, Mitf(+) cells and TRP-2(+) cells appeared, whereas the numbers of TRP 1(+) cells and Kit(+) cells decreased. Some Kit+ cells also expressed Mitf, but no TRP-1 and Mitf double positive cells were seen. In this period, some Mitf(+) cells were labeled with BrdU. The next day, Mitf(+) cells and TRP-2(+) cells increased to the maximum cellularity, and they decreased thereafter and disappeared on the seventh day. An increase of TRP-1(+) cells was first identified on the fifth day after UV exposure, and reached a cellularity four times as great as that of the normal control on the fourteenth day. The subepidermal injection of Kit antibody attenuated the increase of Mitf+ cells and TRP-1(+) cells. These findings suggested that precursor cells that reside in the skin, first differentiated into Mitf(+) and TRP-2(+) melanocytes by the activation of the Kit receptor, then become mature TRP-1(+) melanocytes after UV exposure. PMID- 11407983 TI - Detection of clonal T cell receptor gamma gene rearrangements in cutaneous T cell lymphoma by LightCycler-polymerase chain reaction. AB - Cutaneous T cell lymphoma is thought to be characterized by a monoclonal T cell infiltrate in the skin that can be detected by polymerase chain reaction-based amplification of T cell receptor gamma gene rearrangements. We sought to establish a new, simple, and fast LightCycler-based real-time polymerase chain reaction assay for the detection of monoclonality in cutaneous T cell lymphoma, which was suitable for routine laboratory application. Monoclonal T cell receptor gamma gene rearrangements were detected by polymerase chain reaction with consensus primers using: (i) a thermocycler followed by polyacrylamide gel electrophoresis; (ii) a Light Cycler followed by melting curve analysis; and (iii) a LightCycler and subsequent polyacrylamide gel electrophoresis. The detection limit of monoclonal Jurkat T cells diluted in polyclonal peripheral blood mononuclear cells was: (i) 1--3% by thermocycler--polymerase chain reaction and polyacrylamide gel electrophoresis; (ii) 10% by LightCycler--polymerase chain reaction and melting curve analysis; and (iii) 1% by LightCycler--polymerase chain reaction and polyacrylamide gel electrophoresis. In skin biopsies of 22 cutaneous T cell lymphoma patients, a monoclonal or biclonal T cell infiltrate was detected in: (i) 15 of 22 (68%) by thermocycler--polymerase chain reaction and polyacrylamide gel electrophoresis; (ii) 13 of 22 (59%) by LightCycler- polymerase chain reaction and melting curve analysis; and (iii) 16 of 22 (72%) by LightCycler--polymerase chain reaction and polyacrylamide gel electrophoresis. All three techniques revealed negative results in skin biopsies from 26 patients with benign dermatitis. In conclusion, LightCycler--polymerase chain reaction and melting curve analysis is a fast, simple and specific method to detect monoclonal T cell infiltrates in cutaneous T cell lymphoma. Sensitivity of LightCycler- polymerase chain reaction and polyacrylamide gel electrophoresis is slightly higher compared with sensitivity of thermocycler--polymerase chain reaction and polyacrylamide gel electrophoresis. Melting curve analysis, however, is less sensitive compared with polyacrylamide gel electrophoresis, and in case of negative results of the melting curve analysis, it is recommended to resolve LightCycler--polymerase chain reaction samples by gel electrophoresis. PMID- 11407984 TI - Phenotypic and functional outcome of human monocytes or monocyte-derived dendritic cells in a dermal equivalent. AB - The dermis harbors a true dendritic cell population that could elicit primary allogeneic T cell responses in vitro and contact hypersensitivity reactions in vivo. The origin of dermal dendritic cells remains poorly understood, however. In this study, we analyzed the fate of monocytes or monocyte-derived dendritic cells in a dermal equivalent. Freshly isolated monocytes or monocytes cultured for 6 d with either GM-CSF/IL-4 or GM-CSF/IL-4/TGF-beta 1 (TGF-DC) were seeded in a collagen solution with normal human fibroblasts. The lattices were cultured for 7 -14 d in the presence, or absence, of the exogenous cytokines, before phenotypic and functional studies were performed. Supply of exogenous cytokines allows the appearance of typical CD1a(+)/CD14(-)/CD68(low) dendritic cells with significant allostimulatory property, regardless of the cell type incorporated into the lattices. In cytokine-free conditions, monocytes and GM-CSF/IL-4-derived dendritic cells give rise to a CD1a(-)/CD14(+)/CD68(high) monocyte/macrophage population with no allostimulatory property. When incorporated into the lattices in the absence of exogenous cytokines the TGF-DC express few CD68 and FXIIIa. Interestingly, these cells do not all convert into the CD14(+)/CD1a(-) population. Indeed, a small HLA-DR(+)/CD1a(+)/CD14(-) subset was consistently found, which represents about one-third of the HLA-DR(+) cells. Moreover, TGF-DC recovered from the lattices after culture without cytokines do display a significant allostimulatory function. Thus, in the absence of exogenous cytokines, only Langerhans-cell-like dendritic cells can retain the typical dendritic cell features when inserted in a dermal environment. Taken together, these results may provide evidence supporting an epidermal origin of dermal dendritic cells. PMID- 11407985 TI - Characterization of mouse Frizzled-3 expression in hair follicle development and identification of the human homolog in keratinocytes. AB - Frizzled genes encode a family of Wnt ligand receptors, which have a conserved cysteine-rich Wnt binding domain and include both transmembrane and secreted forms. Work by others has shown that experimental perturbation of Wnt signaling results in aberrant hair formation, hair growth, and hair structure. To date, however, there is no information on the contribution of individual Frizzled proteins to hair development. We now report that Frizzled-3 expression in skin is restricted to the epidermis and to the developing hair follicle. Northern analysis on total mouse skin mRNA revealed a single Frizzled-3 transcript of 3.7 kb. Reverse transcription-polymerase chain reaction and in situ hybridization analysis revealed Frizzled-3 expression in epidermal and hair follicle keratinocytes. Frizzled-3 transcripts are first detected in discrete foci in the developing epidermis of 13 d embryos and later in the hair follicle placodes of 15 d embryos, suggesting a role for this Frizzled isoform in follicle development. In 17 d embryos and 1 d old newborn mice Frizzled-3 expression is limited to suprabasal keratinocytes and is not seen in pelage follicles until 3 d postpartum. In 7 d old neonatal skin, Frizzled-3 is expressed throughout the epidermis and in the outer cell layers of hair follicles. We have also identified the mRNA encoding human Frizzled-3 in epidermal keratinocytes and in the HaCaT keratinocyte cell line. Human Frizzled-3 mRNA encodes a 666 amino acid protein with 97.8% identity to the mouse protein. The human Frizzled-3 gene was mapped using a radiation-hybrid cell line panel to the short arm of chromosome 8 between the markers WI-1172 and WI-8496 near the loci for the Hypotrichosis of Marie Unna and Hairless genes. PMID- 11407986 TI - Patterns of proliferation and apoptosis during murine hair follicle morphogenesis. AB - In this study, we have correlated cutaneous apoptosis and proliferation in neonatal mice during hair follicle morphogenesis. We have applied a novel triple- staining technique that uses Ki67 immunoreactivity as a marker of proliferation as well as TUNEL and Hoechst 33342 staining as apoptosis markers. We have also assessed the immunoreactivity of interleukin-1 beta-converting enzyme, caspase 1, a key enzyme in the execution of apoptosis, and of P-cadherin, which has been suggested as a key adhesion receptor in segregating proliferating keratinocytes. The TUNEL data were systematically compared with high resolution light microscopy and transmission electron microscopy data. Virtually all keratinocytes of the developing hair bud were strongly Ki67(+), suggesting that the hair bud is not an epidermal invagination but primarily the product of localized keratinocyte proliferation. As hair follicle development advanced, three distinct foci of proliferation became apparent: the distal outer root sheath around the hair canal, the mid outer root sheath, and the proximal hair matrix. Of these proliferating hair follicle keratinocytes only defined subsets expressed P cadherin. TUNEL(+) cells in the hair follicle were not found before stage 5 of murine hair follicle morphogenesis. During the early stages of hair follicle development, interleukin-1 beta-converting enzyme immunoreactivity was present on all keratinocytes, but virtually disappeared from the proximal hair follicle epithelium later on. High resolution light microscopy/transmission electron microscopy revealed scattered and clustered apoptotic keratinocytes in all epithelial hair follicle compartments throughout hair follicle development, including its earliest stages. This highlights striking differences in the demarcation of apoptotic hair follicle keratinocytes between the TUNEL technique and high resolution light microscopy/transmission electron microscopy and suggests a role for apoptosis in sculpting the hair follicle even during early hair follicle development. PMID- 11407987 TI - Expression of calcium-binding S100 proteins A4 and A6 in regions of the epithelial sac associated with the onset of hair follicle regeneration. AB - In order to elucidate the onset mechanisms of hair regrowth, we characterized the expression of metastasis and cell-growth-associated calcium-binding S100 proteins in the regenerating follicular epithelium. Hair-cycle-dependent expression of S100A4 and S100A6 was found in the epithelial sac regions (bulge area and hair germ) of mouse pelage follicles. Protein localization of S100A4 was confined to the bulge area, the region where the presumed follicular stem cells reside, during the catagen-telogen-anagen transitional periods, whereas S100A6 protein was distributed throughout the epithelial sac regions during anagen phase. Prior to entering anagen phase, however, both S100 mRNAs were upregulated in the epithelial sac. Despite the induction of extensive cell death in the bulge region after plucking, a new hair cycle was initiated following transcription of S100A6 in the hair germ. Upon wounding stimuli, both the outer root sheath and the basal layer of epidermis expressed S100A6 mRNA prior to hyper-proliferation. Once the epithelial sac was induced to transcribe both S100 genes, resting follicles were concomitantly rejuvenated. These results suggest that S100A4 and S100A6 might play important roles in the activation of stem cells at the onset of follicle regeneration. PMID- 11407988 TI - A novel keratin 5 mutation (K5V186L) in a family with EBS-K: a conservative substitution can lead to development of different disease phenotypes. AB - Epidermolysis bullosa simplex is a hereditary skin blistering disorder caused by mutations in the KRT5 or KRT14 genes. More than 50 different mutations have been described so far. These, and reports of other keratin gene mutations, have highlighted the existence of mutation "hotspots" in keratin proteins at which sequence changes are most likely to be detrimental to protein function. Pathogenic mutations that occur outside these hotspots are usually associated with less severe disease phenotypes. We describe a novel K5 mutation (V186L) that produces a conservative amino acid change (valine to leucine) at position 18 of the 1A helix. The phenotype of this case is unexpectedly severe for the location of the mutation, which lies outside the consensus helix initiation motif mutation hotspot, and other mutations at this position have been associated in Weber- Cockayne (mild) epidermolysis bullosa simplex only. The mutation was confirmed by mismatch-allele-specific polymerase chain reaction and the entire KRT5 coding region was sequenced, but no other changes were identified. De novo K5/K14 (mutant and wild-type) filament assembly in cultured cells was studied to determine the effect of this mutation on filament polymerization and stability. A computer model of the 1A region of the K5/K14 coiled-coil was generated to visualize the structural impact of this mutation and to compare it with an analogous mutation causing mild disease. The results show a high level of concordance between genetic, cell culture and molecular modeling data, suggesting that even a conservative substitution can cause severe dysfunction in a structural protein, depending on the size and structure of the amino acid involved. PMID- 11407989 TI - Expression of a truncated keratin 5 may contribute to severe palmar--plantar hyperkeratosis in epidermolysis bullosa simplex patients. AB - Epidermolysis bullosa simplex are dominant disorders of skin fragility characterized by intraepidermal blistering upon mild mechanical trauma. Skin fragility is caused by expression of either an abnormal keratin 5 or an abnormal keratin 14 protein, which compromises the structure and function of the keratin cytoskeleton of basal cells. We report an epidermolysis bullosa simplex patient with a novel single base substitution (A-->T1414) that changes the lysine residue at amino acid 472 to a non-sense codon (K472X). This change predicts the synthesis of a truncated keratin 5, missing 119 amino acids, including the entire tail domain and the highly conserved KLLEGE motif at the carboxy terminus of the 2B domain of the central rod. Expression of an altered keratin 5, of predicted mass and pI for the product of the K472X allele, was documented by one- and two dimensional western blots of protein extracts from patient skin. Ultrastructural analysis of the patient's nonhyperkeratotic skin was remarkable for basal keratinocytes with dense and irregular keratin filaments proximal to the basement membrane. Keratinocytes, transfected with a cDNA carrying the A-->T1414 non-sense mutation, overexpressed a truncated keratin 5, and showed a disorganized and collapsed keratin filament cytoskeleton. This is the second epidermolysis bullosa simplex patient reported with a premature termination mutation in the KLLEGE motif. The remarkable occurrence of severe palmar--plantar hyperkeratosis in both patients suggests that the keratin 5 tail domain may have unrecognized, but important, normal functions in palmar-plantar tissues. PMID- 11407990 TI - Interleukin-10 promoter polymorphism in psoriasis. AB - Beneficial effects of interleukin-10 therapy and lower endogenous interleukin-10 formation compared with atopic dermatitis and cutaneous T cell lymphomas indicated that interleukin-10 is a key cytokine in psoriasis. The interleukin-10 promoter is highly polymorphic, with two informative microsatellites, interleukin 10.G and interleukin-10.R. In order to understand whether interleukin-10 itself is a predisposing gene for the psoriasis susceptibility we analyzed interleukin 10 promotor polymorphism in patients. The distribution of interleukin-10.G and interleukin10.R microsatellite alleles did not vary between patients (n = 78) and healthy controls (n = 80). In addition, when the psoriasis patients were stratified according to age of onset (younger than 40 y of age, or age 40 and older), no difference in allele distribution was observed; however, a clear differential distribution was revealed at the interleukin10.G locus when patients were stratified according to whether they had a positive family history of psoriasis (p = 0.04). This difference was due to an over-representation of the interleukin10.G13 allele in those patients with familial disease (40.4% vs 19.6%, Chi-square = 7.292, p = 0.007). The positive association of allele interleukin10.G13 with familial psoriasis was especially true when patients with an early onset (< 40 y of age) of the disease were compared with those patients with early onset against a nonfamilial background (39.6% vs 14.5%, Chi-square = 8.959, p = 0.003). Patients with age-of-onset of less than 40 were 4-fold [odds ratio = 3.85 (1.55--9.62)] more likely to have a psoriatic family background if they carried this interleukin10.G13 allele. These data suggest that the interleukin-10 locus contributes to the heritability of psoriasis susceptibility. PMID- 11407991 TI - Safety and pharmacokinetics of naked plasmid DNA in the skin: studies on dissemination and ectopic expression. AB - Gene therapy using naked DNA injected into muscle and skin is increasingly being used for vaccination and treatment purposes. Favorably, naked plasmid DNA does not exhibit the various limitations inherent to viral vectors, such as the elicitation of adverse immune responses and the risk of insertional mutagenesis. In order to assess the distribution and safety of naked plasmid DNA in a relevant animal model, we analyzed if intracutaneously injected plasmid DNA was transported to other organs and if ectopic expression occurred. When a "superdose" of a marker plasmid was injected intradermally, most organs were found transiently to contain the plasmid DNA for several days, whereas integration into the host genome was not detected. With the exception of ovary, however, mRNA expression only occurred in the skin, regional lymph nodes, and muscular tissues. From a safety standpoint, skin gene therapy with naked plasmid DNA can be considered safe due to the rapid biodegradation of plasmid DNA and the exclusive and transient expression of foreign genes in tissues known to take up DNA. PMID- 11407992 TI - Detection of differentially regulated genes in keratinocytes by cDNA array hybridization: Hsp27 and other novel players in response to artificial ultraviolet radiation. AB - cDNA array technology was used to identify novel genes participating in the ultraviolet response of cultured human keratinocytes. cDNA arrays representing more than 50,000 different cDNA clones were hybridized with complex probes generated by SMART-polymerase chain reaction amplification of 150 ng of total RNA extracted 24 h after ultraviolet irradiation. Fifty-one clones with differential hybridization signals were detected, representing 19 different sequences; 10 known genes (seven ultraviolet induced, three ultraviolet suppressed) and further nine expressed sequence tags of unknown genes. In seven of 10 genes the data from cDNA arrays probed with SMART-cDNA could be confirmed by northern blot analysis. The 27 kDa heat shock protein mRNA was induced. Keratins 6 and 17, markers for the hyperproliferative status of keratinocytes, were among the ultraviolet suppressed genes. The change of expression profile of keratins indicates a differentiation towards a phenotype of keratinocytes present in the suprabasal layers of the epidermis. These mechanisms may contribute to the ultraviolet protective function of the epidermis and to the anti-proliferative action of ultraviolet in the therapy of psoriasis. We also detected an induction of adenylyl cyclase associated protein and the suppression of G(s)alpha (a stimulating subunit of the trimeric membrane bound GTPase). PMID- 11407993 TI - High macrophage migration inhibitory factor (MIF) serum levels associated with extended psoriasis. PMID- 11407994 TI - A novel leucine to valine mutation in residue 7 of the helix initiation motif of keratin10 leads to bullous congenital ichthyosiform erythroderma. PMID- 11407995 TI - Compound heterozygous TGM1 mutations including a novel missense mutation L204Q in a mild form of lamellar ichthyosis. PMID- 11407996 TI - The (CTG)n polymorphism in the NOTCH4 gene is not associated with schizophrenia in Japanese individuals. AB - BACKGROUND: The human NOTCH4 gene is a candidate gene for schizophrenia due to its chromosomal location and neurobiological roles. In a British linkage study, NOTCH4 gene polymorphisms were highly associated with schizophrenia. In a Japanese case-control association study, however, these polymorphisms did not show significant associations with schizophrenia. We conducted a case-control study with Japanese subjects to explore an association between the triplet repeat polymorphism in the NOTCH4 gene and schizophrenia, including subtypes of schizophrenia, longitudinal disease course characteristics, and a positive family history for psychoses. METHODS: We examined the (CTG)n repeat polymorphism in the NOTCH4 gene among 100 healthy Japanese individuals and 102 patients with schizophrenia (22 paranoid, 38 disorganized, 29 residual, 64 episodic, 31 continuous, 42 with prominent negative symptoms, and 46 with positive family histories) using a polymerase chain reaction-based, single-strand conformational polymorphism analysis. RESULTS: Five different alleles consisting of 6, 9, 10, 11, and 13 repeats of CTG (Leu) in patients with schizophrenia, and 4 alleles consisting of 6, 9, 10, and 11 repeats in controls were found. No significant differences in genotype or allele frequencies of repeat numbers were found between controls and patients. In addition, there were no associations between the polymorphism and schizophrenia subtypes, longitudinal disease course characteristics, or positive family history of the patients. CONCLUSIONS: Our data suggest a lack of association between the NOTCH4 gene triplet repeat polymorphism and schizophrenia in Japanese individuals. PMID- 11407997 TI - Decreases in blood perfusion of the anterior cingulate gyri in Anorexia Nervosa Restricters assessed by SPECT image analysis. AB - BACKGROUND: It is possible that psychopathological differences exist between the restricting and bulimic forms of anorexia nervosa. We investigated localized differences of brain blood flow of anorexia nervosa patients using SPECT image analysis with statistic parametric mapping (SPM) in an attempt to link brain blood flow patterns to neurophysiologic characteristics. METHODS: The subjects enrolled in this study included the following three groups: pure restrictor anorexics (AN-R), anorexic bulimics (AN-BP), and healthy volunteers (HV). All images were transformed into the standard anatomical space of the stereotactic brain atlas, then smoothed. After statistical analysis of each brain image, the relationships among images were evaluated. RESULTS: SPM analysis of the SPECT images revealed that the blood flow of frontal area mainly containing bilateral anterior cingulate gyri (ACC) was significantly decreased in the AN-R group compared to the AN-BP and HV groups. CONCLUSIONS: These findings suggest that some localized functions of the ACCare possibly relevant to the psychopathological aspects of AN-R. PMID- 11407998 TI - Randomized placebo-controlled trial of long-term treatment with sibutramine in mild to moderate obesity. AB - OBJECTIVE: The researchers assessed the long-term weight reduction efficacy, tolerability, and safety of sibutramine used once daily in conjunction with behavior modification to treat mild to moderate obesity. STUDY DESIGN: This was a double-blind randomized placebo-controlled parallel-group comparative study of sibutramine 10 mg or 15 mg (or placebo) once daily for 1 year, given with dietary advice. POPULATION: A total of 485 obese men and women with uncomplicated obesity were included (mean age=42 years, mean body mass index=32.7 kg/m2). OUTCOMES MEASURED: The outcomes were mean weight loss, percentage losing more than 5% or 10% of their body weight, and adverse drug effects. RESULTS: Among patients completing the study, those taking sibutramine 10 mg or 15 mg had greater mean weight loss compared with placebo at 12-month assessment (P < or = .001). Changes in body weight from baseline to end point were -1.6 kg for those taking placebo, 4.4 kg for those taking sibutramine 10 mg (P < or =.01, last observation carried forward [LOCF]), and -6.4 kg for those taking sibutramine 15 mg (P < or =.001, LOCF). For placebo patients, 20% lost 5% or more of their body weight compared with 39% of patients taking sibutramine 10 mg and 57% taking sibutramine 15 mg. Only 7% of the patients taking placebo lost 10% or more of their body weight, compared with 19% taking sibutramine 10 mg and 34% taking sibutramine 15 mg (P <.001 for both 10 mg and 15 mg vs placebo, and for both > or =5% and > or =10%). CONCLUSIONS: Sibutramine 10 mg or 15 mg once daily given with dietary advice produces and maintains statistically and clinically significantly greater weight loss than dietary advice alone (placebo) throughout a 12-month treatment period, and is safe and well tolerated. PMID- 11407999 TI - Serum cystatin C in patients with myeloma. AB - BACKGROUND: Cystatin C is a low molecular weight protein thought to be synthesised by all nucleated cells and freely filtered by the kidney. It has been proposed as a marker for GFR; however, it has been suggested that there may be limitations to its use, because it may be over-expressed in some tumour cells and the abnormal tissue growth may also lead to an increased circulating level. METHODS: We investigated the serum cystatin C levels in 60 patients with myeloma, comparing results with those for serum creatinine, beta(2)-microglobulin and the paraprotein concentration. RESULTS: We found no correlation between cystatin C and the paraprotein concentration in these patients. CONCLUSION: These results suggest that disease burden does not correlate to the circulating level of cystatin C in patients with myeloma. PMID- 11408000 TI - Multiplex PCR assay for screening deletions in the low density lipoprotein receptor gene. AB - BACKGROUND: In different populations of the world, more than 150 genetic alterations of the LDL receptor gene have been identified; each of which can result in hypercholesterolaemia, but no hot spots in the gene were detected so far. Because of the existence of very variable genetic alterations in different ethnic communities, none of the assays developed for screening mutations/deletions in a population defined can be adapted to study the possible genetic defects. The present study was designed to develop a new, multiplex PCR based, molecular biological method to screen the whole coding region of the LDL receptor gene. METHODS: Using primer pairs completely flanking the promoter and the entire exonal region, in the PCR reactions 83-386-bp long, DNA sequences were synthesised in seven different reaction mixtures. The reaction conditions of the multiplex PCR system were optimised in order to synthesise all exons and the promoter region of the gene using only two annealing temperatures. The products could be visualised separately by agarose gel electrophoresis/ethidium bromide staining. RESULTS: A rapid, effective test enabling the screening of DNA alterations in the entire LDL receptor gene was developed. Using this simple multiplex PCR assay, deletions affecting more than 10 bp in any part of the gene can be easily detected by a single agarose gel electrophoresis. CONCLUSIONS: The simplicity, specificity and versatility of the assay make it suitable system for routine screening of LDL receptor gene mutations in large population samples. This PCR assay can be recommended for screening of LDL-RG deletions in populations or groups at high risk for cardiovascular diseases. PMID- 11408001 TI - Analysis of Gq protein alpha subunit mRNA expression in human monocytes: relevance of the purification step. AB - BACKGROUND: Galphaq is a member of the Gq family of G proteins, which by stimulating the phospholipase Cbeta (PLCbeta)-IP(3)-Ca(++) mediated intracellular signaling systems controls some of the most fundamental cardiovascular cellular processes. The study of Galphaq is complicated by the presence of a pseudogene in human DNA, with signficant homology to the mRNA encoding the alpha subunit of Gq protein. The presence of this pseudogene will cause problems when the analysis of the Galphaq gene expression is based solely on RT-PCR. In this study, we report a simple method for avoiding unwanted amplification of the Galphaq pseudogene and the use of human monocytes as a readily available source for examining Galphaq. METHODS: RT-PCR was carried out on RNA extracted from peripheral blood monocytes of 10 normal subjects using specific primers for Galphaq. RESULTS: When several subjects' Galphaq was examined, the authentic Galphaq mRNA amplification product levels, as a ratio to unpurified pseudogene containing amplification products, declined by up to approximately 70%. CONCLUSION: Given the importance of Gq protein in cardiovascular signal transduction, it is fundamental to provide a reliable assessment of G alpha q gene expression. In addition to accurately assessing Galphaq levels, the use of circulating human monocytes as a useful source of Galphaq for investigating mechanisms involved in the regulation of vascular tone is shown. PMID- 11408002 TI - Platelet antioxidant enzymes in insulin-dependent diabetes mellitus. AB - BACKGROUND: The measurement of the peroxidase scavenging system represented by the activities of superoxide dismutase (SOD), catalase and glutathione peroxidase (GSH-Px) in blood cells of diabetic patients has, in the past, given equivocal results. Likewise, the role of these intracellular enzymatic scavengers against the oxidative stress of diabetes-associated microangiopathic complications is unknown. METHODS: Choosing platelets as cell model (as commonly done in previous studies), the aim of this study was to relate the platelet content of SOD, catalase and GSH-Px to the presence of diabetes, as well as to the presence of nephropathy and retinopathy in 35 insulin-dependent diabetic patients, as compared to 10 age-matched control subjects. RESULTS: The enzymatic activities were not changed in diabetic patients in comparison with healthy controls. After stratifying patients according to presence of nephropathy (24-h urinary albumin excretion rate persistently > or =20 microg min(-1)) or retinopathy, the group of albuminuric patients was characterized by a significant decrease in SOD activity as compared to those in the normoalbuminuric range (4.36+/-1.06 vs. 6.81+/-2.26 mU 10(-9) platelets; p=0.01). Catalase and GSH-Px did not change. No modification in platelet enzyme activities has been found in diabetic subjects with retinopathy. CONCLUSIONS: These results suggest that diabetic nephropathy, at least in its early stage, may be related to an altered redox state of platelets, as tested by the reduction in SOD activity, thus, indicating that the renal damage in these patients may be associated to a selective increase in platelet susceptibility to variation in the redox state. PMID- 11408003 TI - Seasonal and biological variation of urinary epinephrine, norepinephrine, and cortisol in healthy women. AB - BACKGROUND: There is a significant circadian and seasonal periodicity in various endocrine functions. The present study describes the within-day and seasonal fluctuation for urinary catecholamines and cortisol and estimates the within- (CV(i)) and between-subject (CV(g)) coefficients of variation for healthy women undertaking their routine work. In addition, index of individuality (I(i)) and power calculations were derived. METHODS: Eleven healthy females undertaking their routine life-style at work participated in the study. Each subject collected six samples during 24 h 15 days over a year, giving a total number of 990 samples. Using a random effect analysis of variance, we estimated CV(g) and total within-subject variation (CV(ti)), i.e. combined within-subject and analytical variation, from logarithmically transformed data. Analytical variation was subtracted from CV(ti) to give CV(i). CV(i) was estimated from samples collected monthly during 1 year (CV(iy)), weekly during 1 month (CV(im)), and six to eight times/day (CV(id)). RESULTS: A seasonal variation was demonstrated for excretion of epinephrine, norepinephrine, and cortisol standardized with creatinine. Concentrations of urinary epinephrine were higher during June and July compared to the rest of the year, whereas concentrations of urinary cortisol were higher during December and January compared to the rest of the year. Excretion of norepinephrine was lower during working hours and higher during hours off work for June and July compared to the rest of the year. There was a high within- and between-subject variation, which could not be explained by menstrual cycle, behavioral, emotional, or cognitive stress reactions. CONCLUSIONS: Despite high biological variation a reasonably low sample size, e.g. 10-50 individuals, is adequate for practical applicability, i.e. studying differences above 150%. The present study recommends to include the sampling time in the statistical evaluation of data and to be aware of the changes in diurnal variations over seasons. When single measurements are to be evaluated, reference intervals are recommended. PMID- 11408004 TI - Expression of cathepsin L and its endogenous inhibitors in immortal and transformed fibroblasts. AB - METHODS: The activities of cathepsin L and its endogenous inhibitors were analyzed in rat embryo fibroblasts, immortalized and transformed by different genes. RESULTS: Regardless of the transfecting agent used (DNA of adenovirus SA7 or polyomavirus LT gene), the immortal cells showed an increase in the cathepsin L activity (in both lysates and conditioned media) compared to primary fibroblasts. Transformed cells exhibited either an increase (with c-Ha-ras gene) or decrease (with E7 HPV gene) in cathepsin L activity in lysates as opposed to immortal cells. CONCLUSIONS: The data are suggestive of alterations in the trafficking of cathepsin L upon fibroblast transfection with polyomavirus LT gene and E7 HPV gene. An endogenous inhibitor(s) of cysteine proteinase was found in conditioned media, but not in lysates, of all cell cultures studied and its activity in normal fibroblasts was higher than in media of immortal and transformed cells. PMID- 11408005 TI - The association of serum lipoprotein(a) levels, apolipoprotein(a) size and (TTTTA)(n) polymorphism with coronary heart disease. AB - BACKGROUND: The association between lipoprotein(a) levels, apolipoprotein(a) size and the (TTTTA)(n) polymorphism which is located in the 5' non-coding region of the apo(a) gene was studied in 263 patients with severe coronary heart disease and 97 healthy subjects. METHODS: Lp(a) levels were measured by ELISA, apo(a) isoform size was determined by SDS-agarose gel electrophoresis, and analysis of the (TTTTA)(n) was carried out by PCR. For statistical calculation, both groups were divided into low (at least one apo(a) isoform with < or = 22 Kringle IV) and high (both isoforms with >22 KIV) apo(a) isoform sizes, and into low number (<10 in both alleles) and high number of (> or =10 at least one allele) TTTTA repeats. RESULTS: Lp(a) levels were higher (P=0.007), apo(a) isoforms size < or =22 KIV and TTTTA repeats > or = 10 were more frequent (P=0.007 and 0.01) in cases than in controls. Lp(a) levels were found to be increased with low apo(a) weight in both groups (both P<0.0001). In multivariate logistic regression analysis, only the Lp(a) levels (P=0.005) and (TTTTA)(n) polymorphism (P=0.002) were found to be significantly associated with CHD. CONCLUSION: Nevertheless, these results indicate that in CHD patients the (TTTTA)(n) polymorphism has an effect on Lp(a) levels which is independent of the apo(a) size. PMID- 11408006 TI - Effect of anticoagulants on plasma homocysteine determination. AB - BACKGROUND: Waiting temperature before centrifugation and anticoagulants used, markedly effect total homocysteine concentrations. The aim of this study was to investigate the effect of different anticoagulants and temperature on plasma homocysteine levels. METHODS: We studied total homocysteine concentrations in 23 healthy subjects. Blood was drawn in K(3)EDTA, sodium citrate- or sodium fluoride containing tubes, and kept at 0 degrees C or 22 degrees C for 3 h. Total homocysteine measurements were performed with fluorescence polarization immunoassay (FPIA) method. We compared all results with baseline EDTA values (samples put on crushed ice and centrifuged immediately) recommended in literature for reference handling. RESULTS: At 22 degrees C, the tubes containing sodium citrate and sodium fluoride showed significantly higher total homocysteine concentrations than their respective baseline values (p=0.000). However, sodium fluoride tubes were not significantly different than baseline EDTA levels. Waiting 3 h at 0 degrees C did not effect sodium citrate and EDTA plasma total homocysteine concentrations when compared to baseline EDTA, but sodium fluoride containing plasma levels were significantly decreased (p=0.000). CONCLUSIONS: According to our results, the most available and practical temperature and anticoagulant for total homocysteine determination is sodium fluoride at room temperature up to 3 h. PMID- 11408007 TI - The comparison of the ability of monoclonal antibodies directed to different proteins (human IgG, human myoglobin and HRP) and bispecific antibodies derived thereof to bind antigens immobilized on a surface of a solid phase. AB - BACKGROUND: Bindings of mouse monoclonal antibodies (mAbs) and affinity purified bispecific antibodies (bAbs), derived thereof, to antigens adsorbed on immunoplates have been compared, using ELISA and RIA methods. METHODS: The analysed panel of antibodies included mAbs specific to human myoglobin (Mb), human IgG (hIgG) and horseradish peroxidase (HRP) and biologically produced bAbs with double specificity to Mb and HRP, and to hIgG and HRP. RESULTS: The degree of difference between different mAbs and corresponding bAbs varied markedly from antibody to antibody, depending on whether the parental mAbs could bind immobilized antigens bivalently. The observed equilibrium binding constant (K(obs)) for anti-HRP mAbs was 21-38 times higher that of anti-HRP site of bAbs (anti-hIgG/HRP or anti-Mb/HRP, respectively), due to bivalent binding of mAbs. Anti-Mb mAbs also bound bivalently with immobilized Mb. On the contrary, anti hIgG mAbs bound monovalently with immobilized hIgG in the same conditions. The avidity of anti-Mb/HRP bAbs increased, if both antigens were simultaneously adsorbed on a solid phase. CONCLUSIONS: The obtained data indicate that the use of bAbs in heterogeneous immunoassays instead of traditional mAb-enzyme conjugates hardly can provide the significant gain in assay performance if parental mAbs bind bivalently. PMID- 11408008 TI - alpha-Amylase expressed in human liver is encoded by the AMY-2B gene identified in tumorous tissues. AB - BACKGROUND: An alpha-Amylase in human liver is detected with an anti-human salivary amylase antibody, but the enzyme activity is very low. We previously found that the rat liver contained an amylase which differed from the enzyme of mice. In this study, we characterized the human liver amylases biochemically and immunohistochermically. METHODS AND RESULTS: Although the amylase activity of human liver was much lower than that of rat, protein moiety and sugar chains of the human amylase were identified as similar to the rat liver enzyme with an anti human salivary amylase antibody and by concanavalin A (Con A) affinity chromatography. Liver amylases from human and rat were the same size, 50 kDa, on Western blot analysis and had the same isoelectric points. The cytoplasm of hepatocytes was moderately stained immunohistochemically with the anti-human salivary amylase antibody. Intrahepatic bile ducts were also stained weak-to moderately. RT-PCR, with a specific primer for the consensus sequence of human amylases, amplified a single 474-bp product from the human liver total RNA. The PCR product was sequenced and referred to the homology. Thirteen bases in the 434 bp fragment of the human liver amylase differed from the corresponding region of the AMY-1 gene transcript and the deduced amino acid sequence differed at five residues. The human liver amylase cDNA sequence was identical to the corresponding cDNA of the AMY-2B, which was known to expressed in tumorous tissues. In situ hybridization revealed the expression of AMY-2B mRNA in non tumorous human liver. CONCLUSIONS: The present results suggest the possibility that a novel amylase detected in tumorous tissues and encoded by the AMY-2B gene is a liver-specific amylase expressed in the human liver. PMID- 11408009 TI - Novel findings in inhibition of mast cell-dependent immediate-type cutaneous reactions by Gahmi-Shini-San. AB - This report describes an inhibitory effect of Gahmi-Shini-San (GSS) on mast cell mediated immediate-type allergic reactions. GSS is an Oriental herbal medication, which has been successfully used in Korea for the treatment of allergic disorders, mainly skin anaphylactic diseases. GSS inhibited the ear swelling response induced by intradermal injection of compound 48/80 in a mouse model on a concentration-dependent basis. The mast cells in mouse ear tissue were stained by alcian blue/nuclear fast red. GSS significantly inhibited the compound 48/80 induced degranulation from mast cells in ear tissue. GSS dose-dependently inhibited the histamine release from the rat peritoneal mast cells by compound 48/80. We also studied the effect of GSS on mast cell-dependent passive cutaneous anaphylaxis activated by dinitrophenyl IgE antibody. GSS showed inhibition of passive cutaneous anaphylaxis following oral administration. These results indicated that GSS has inhibitory effect on mast cell-dependent immediate type cutaneous reactions. PMID- 11408010 TI - The Fab fragment of anti-digoxin antibody (digibind) binds digitoxin-like immunoreactive components of Chinese medicine Chan Su: monitoring the effect by measuring free digitoxin. AB - Chan Su, a Chinese medicine prepared from the skin glands of Chinese toads, is used in the treatment of cardiovascular diseases. Severe toxicity and even death has been reported from overdose with Chan Su. The cardiotonic effect of Chan Su is attributed to bufadienolides, which also have apparent digitoxin activity. We demonstrated that these components of Chan Su could be neutralized by digibind, both in vitro and in vivo. For in vitro experiments, we supplemented drug-free serum pools with aqueous extract of Chan Su. Then, to aliquots of serum pool containing Chan Su, various amounts of digibind (10, 25 or 50 microg/ml of serum) were added. After incubation, total and free digitoxin concentrations (in the protein-free ultrafiltrate) were measured using the fluorescence polarization immunoassay (FPIA) and a FLX/TDx analyzer. For in vivo experiments, mice were fed with Chan Su by gavage. After 45 min, 200 microg of digibind was administered by injection. Fifteen minutes after injection, blood was collected for analysis of total and free apparent digitoxin activities. We observed complete removal of apparent digitoxin activity from protein-free ultrafiltrate both in vitro and in vivo by digibind, indicating that digibind successfully binds Chan Su. We conclude that digibind neutralizes Chan Su, and measuring the free digitoxin concentrations can monitor such an effect. PMID- 11408011 TI - Antioxidant status and lipid peroxidation in premenstrual syndrome: a preliminary study. PMID- 11408013 TI - Immunopathogenesis of hepatitis C viral infection: Th1/Th2 responses and the role of cytokines. AB - Hepatitis C virus is a common cause of hepatocellular injury that is associated with complex and vigorous immunologic mechanisms. Both humoral and cell-mediated immune responses participate in the host defense against hepatitis C viral infection, but there is increasing recognition of the roles played by the cell mediated response, and in particular the cytokine system, in the immunopathogenesis of chronic hepatitis C. The cell-mediated response depends on cytotoxic and helper T-cell activity, and functions through the actions of cytokines to regulate macrophages, natural killer cells, and antiviral cellular proteins. Cytokines produced in the liver are essential in defending the host against hepatitis C invasion, but they have also been implicated in the hepatocellular injury seen in the majority of chronically infected patients. Cytokines are thought to be involved in the pathogenesis of hepatitis C under conditions where the virus can mutate effectively and evade T-cell immune defense mechanisms. Persistent infection upsets the balance between immunostimulatory and inhibitory cytokines which can prolong inflammation and lead to necrosis, fibrosis, and chronic liver disease. PMID- 11408014 TI - Cytokines as predictors for sustained response and as markers for immunomodulation in patients with chronic hepatitis C. AB - OBJECTIVES: (i) To characterize serum cytokine levels of tumor necrosis factor alpha (TNF alpha), interleukin 6 (IL 6), IL 8 and IL 12 in non-cirrhotic patients with chronic hepatitis C, (ii) to correlate the levels of these cytokines with the degree of the disease at the basal level, (iii) to correlate these levels with the response to therapy, (iv) to compare profiles of cytokines in monotherapy (MT) versus combination therapy (CT), and (v) to compare the immunomodulatory effects of MT versus CT. DESIGN AND METHODS: 47 patients were enrolled in the study. The controls were 120 volunteers (recruited from students and staff) that did not present HCV RNA positive and were not known to suffer any other metabolic disease. Thirty patients formed the other group of controls, with alcoholic liver disease (ALD). Serum cytokine levels were assessed using enzyme linked immunosorbent assay (ELISA). RESULTS: The sustained responders (SRs) have basal values much lower than relapsed responders (RRs) and non-responders (NRs) regardless of the therapy. CONCLUSIONS: Cytokines can be used as non-invasive markers for sustained response and as monitors for the outcome of therapy. PMID- 11408015 TI - Cytokine network in nonresponding chronic hepatitis C patients with genotype 1: role of triple therapy with interferon alpha, ribavirin, and ursodeoxycholate. AB - OBJECTIVE: (i) to characterize the profile of tumor necrosis factor alpha (TNF alpha), interleukin-6 (IL-6), IL 10, Fas-ligand and transforming growth factor beta (TGF beta), chronic hepatitis C (HCV) patients with genotype 1; (ii) to determine the influence of triple therapy (TT) with interferon alpha (IFN alpha) + ribavirin + ursodeoxycholic acid on these cytokines and (iii) to establish the relationship between the pro-inflammatory cytokines and the outcome of treatment. DESIGN AND METHODS: 22 patients infected with HCV-genotype 1 a/b and non responsive to IFN-alpha monotherapy were enrolled in the TT. The controls were 49 HCV naive patients with genotype 1 a/b. Cytokine levels were measured using enzyme-linked immunosorbent assay (ELISA). RESULTS: The baseline TNF alpha values (pg/mL) in the sustained responders (SRs) (63+/-3) were significantly lower than non-responders (NRs) (140+/-16) (p < 0.001). Baseline Fas (ng/mL) levels were also lower in SRs (4.3+/-0.2) than NRs (5.4+/-0.4) (p < 0.05). CONCLUSIONS: Fas and TNF alpha may be used as serological markers of inflammation and effectiveness of therapy. PMID- 11408016 TI - Increased serum levels of macrophage migration inhibitory factor in alcoholic liver diseases and their expression in liver tissues. AB - OBJECTIVES: To study the role of macrophage migration inhibitory factor (MIF) in the pathogenesis of alcoholic liver diseases. DESIGN AND METHODS: The levels of MIF in the sera were estimated by an enzyme-linked immunosorbent assay in 13 patients with alcoholic hepatitis (ALH), 9 patients with alcoholic cirrhosis (ALC) and 26 normal controls. MIF was localized in the liver specimens by immunohistochemistry. RESULTS: The mean levels of MIF in the sera were significantly higher in ALH and ALC compared with the normal controls (p < 0.05). Serial observations revealed a relationship between serum MIF levels and the serum transaminase levels. MIF was expressed by the hepatocytes and by the infiltrating cells around the site of accumulation of neutrophils and ballooned hepatocytes in ALH. CONCLUSIONS: This is the first report on MIF in human alcoholic liver diseases, and the data suggest that MIF may be related to abnormal cytokine homeostasis in ALH. PMID- 11408017 TI - Accumulation of macrophages in primary sclerosing cholangitis. AB - OBJECTIVES: To determine what changes are occurring in patients with primary sclerosing cholangitis (PSC) by examining perisinusoidal macrophages (Kupffer cells) in liver biopsies; 2-to measure transforming growth factor beta (TGFbeta) as a marker of fibrosis in these patients. DESIGN AND METHODS: Transmission electron microscopy and immunohistochemistry of 15 PSC, 26 primary biliary cirrhosis (PBC), 30 alcoholic liver disease (ALD) and 51 with normal histology was used. Five PSC, 30 ALD and 120 normal volunteers were sampled for serum levels of TGFbeta. RESULTS: There was a three-fold increase in relative numbers of Kupffer cells in PSC compared to PBC and to patients whose livers had normal histology. In PSC there was an accumulation of perisinusoidal macrophages, which was not associated with focal necrosis or with cholestasis. The levels of TGFbeta in PSC were 54 +/- 2 in cirrhotic versus 34 +/- 5 in non-cirrhotic patients (p < 0.005). CONCLUSION: The persistent activation of these macrophages may lead to the chronic release of TGFbeta and contribute to chronic inflammation, fibrosis and cirrhosis. PMID- 11408018 TI - Ethanol inhibits the JAK-STAT signaling pathway in freshly isolated rat hepatocytes but not in cultured hepatocytes or HepG2 cells: evidence for a lack of involvement of ethanol metabolism. AB - OBJECTIVES: To understand the molecular mechanism underlying alcoholic liver injury, effects of acute ethanol on the Janus kinase-signal transducer and activator transcription factor (JAK-STAT) signaling in hepatic cells were studied. DESIGNS AND METHODS: Effects of acute ethanol on the JAK-STAT signaling in freshly isolated, cultured rat hepatocytes, and HepG2 cells were explored. RESULTS: Acute ethanol exposure inhibited IL-6- or IFN-activated STAT in freshly isolated hepatocytes but not in cultured hepatocytes, HepG2 cells, or HepG2 cells transfected with alcohol dehydrogenase (ADH) or cytochrome P450(2E1). The inhibitory action of ethanol in freshly isolated hepatocytes was not antagonized by the ADH inhibitor 4-methylpyrazole (4-MP). Acute exposure of hepatocytes to acetaldehyde or hydrogen peroxide did not suppress STAT activation. Further studies indicated that the loss of response to the inhibitory effect of ethanol was not due to hepatocyte proliferation and collagen contact. CONCLUSIONS: Freshly isolated hepatocytes are more susceptible to the inhibitory action of ethanol on the JAK-STAT signaling than cultured hepatocytes or HepG2 cells, which may be implicated in pathogenesis and progression of alcoholic liver disease. PMID- 11408019 TI - CYP2E1-mediated modulation of valproic acid-induced hepatocytotoxicity. AB - OBJECTIVES: To determine the cytotoxicity of valproic acid (VPA) and its metabolite, 4-ene-valproic acid (4-ene-VPA) in human hepatoblastoma cells (Hep G2), and to study the modulatory effect of cytochrome P450 2E1 induction in this model. METHODS: Cells were exposed to VPA or 4-ene-VPA in the presence of either ethanol (EtOH), or EtOH combined with disulphiram (DS). Some cells were exposed to alpha-fluoro-VPA or to alpha-fluoro-4-ene-VPA in the absence of CYP2E1 inducers. Apoptosis and necrosis were measured by analyzing 6000 cells per sample using transmission electron microscopy, while cytokine release and apoptosis were quantitated by ELISA. RESULTS: VPA + EtOH increased VPA cytotoxicity. 4-ene-VPA + EtOH significantly increased toxicity, while DS + EtOH significantly reduced this toxicity. Alpha-fluorinated analogues reduced cytotoxicity compared to the corresponding VPA compounds. Neither VPA nor alpha-fluorinated VPA increased the release of IL-6 or TNF-alpha in media. A significant increase in the release of TNF-alpha was observed in cells exposed to 4-ene-VPA that further increased with EtOH exposure. CONCLUSIONS: Cells exposed to 4-ene-VPA experience greater cytotoxicity than those treated with VPA. Cytochrome P450 2E1 inducers enhance toxicity in VPA-exposed cells, while alpha-fluorination of VPA diminishes cytotoxicity by directly interfering with the beta-oxidation of the 4-ene-VPA metabolite. PMID- 11408020 TI - Signaling for ethanol-induced apoptosis and repair in vitro. AB - OBJECTIVES: To evaluate whether caspases are involved in ethanol (EtOH)-induced apoptosis and if polyenylphosphatidylcholine (PPC) affects apoptosis, in vitro in Hep G2 cells. METHODS: Cells were treated with 100 mmol/L EtOH for 24 h and with 2 doses of 100 mmol/L EtOH (1/24 h) in the presence of absence of 20 mmol/L of PPC or 50 micromol/L caspase 3 inhibitor (IDN). Cells were analyzed for apoptosis by transmission electron microscopy (TEM) 6000 cells/treatment, DNA fragmentation by ELISA and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate (T dt-mediated d-UTP) nick-end-labeling, TUNEL. RESULTS: 100 mmol/L dose of EtOH resulted in 22 +/- 2.5% (p < 0.001) apoptosis (vs. control). Two consecutive doses of 100 mmol/L EtOH for 24 h each caused 36 +/- 3.0% (p < 0.001 vs. control and p < 0.05 vs. one dose). PPC significantly reduced apoptosis (vs. non exposed to PPC): 100 mmol/L -12 +/- 1.5% (p < 0.05) and 2 x 10(-)(0) mmol/L 20 +/- 2.0% (p < 0.001). Pretreatment with 50 micromol caspase inhibitor reduced EtOH-induced apoptosis in a similar proportion. CONCLUSIONS: PPC downregulates EtOH-apoptosis by a mechanism similar to caspase inhibition. PMID- 11408021 TI - The toxicity of Callilepis laureola, a South African traditional herbal medicine. AB - OBJECTIVES: To review the literature on the toxicity of Callilepis laureola, and to assess the cytotoxicity of C. laureola in human hepatoblastoma Hep G2 cells in vitro. DESIGN AND METHODS: Cells were incubated for up to 48 h in the presence of increasing concentrations of an aqueous extract of C. laureola (0.3-13.3 mg/mL). Cytotoxicity was quantitated spectrophotometrically by the metabolism of the tetrazolium dye MTT. Cytoviability of the control cells was considered to be 100%. RESULTS: C. laureola produced cytotoxicity in a concentration-dependent manner. Cytotoxicity was significant at all concentrations tested (0.3-2.5 mg/mL, p < 0.05 vs. controls and 3.3-13.3 mg/mL, p < 0.0001 vs. controls). After 6 h, 100% toxicity was observed at a concentration of 6.7 mg/mL. CONCLUSION: C. laureola causes significant cytotoxicity in Hep G2 cells in vitro. These findings are in accordance with the observed hepatotoxicity in clinical cases of C. laureola poisoning. PMID- 11408022 TI - The role of interleukin-10 in severe acute alcoholic hepatitis. PMID- 11408023 TI - Preliminary investigation of near-infrared spectroscopic measurements of urea, creatinine, glucose, protein, and ketone in urine. AB - OBJECTIVE: We investigated the use of near-infrared spectroscopy as an analytical tool to quantify concentrations of urea, creatinine, glucose, ketone, and protein in urine. DESIGN AND METHODS: FT-IR spectroscopy in conjunction with a polynomial based spectral smoothing method was applied to urine specimens. A partial factorial experimental design was employed to collect spectra using normal and spiked urine samples. RESULTS: Our results show that the spectral signatures of urea, creatinine, glucose, ketone, and protein in the 1350 to 1800 nm and 2050 to 2375 nm range are sufficiently strong and unique for accurate measurements. CONCLUSIONS: The accuracy of near infrared for quantifying concentrations of urea and creatinine is only slightly less than our selected reference methods. Glucose, ketone and protein are sufficiently accurate to be useful as a screening tool for wellness. The method successfully accounts for biologic matrix variation. The advantages of near-infrared analysis are (1) no reagents, (2) ease of sample preparation, (3) speed, and (4) the ability to quantify multiple analytes with one spectra. PMID- 11408024 TI - The relationship of oxidative DNA damage marker 8-hydroxydeoxyguanosine and glycoxidative damage marker pentosidine. AB - OBJECTIVES: 8-hydroxydeoxyguanosine (8-OHdG) is a biomarker of oxidative DNA damage. Pentosidine is a biomarker of glycoxidation reaction. In this study, we investigated relationships among 8-OHdG, pentosidine and age. DESIGN AND METHODS: We determined the urinary concentrations of 8-OHdG and pentosidine in adults with mild hypercholesterolemia or/and mild hypertension (hypercholesterolemia group, n = 31; hypertension group, n = 25; hypercholesterolemia and hypertension group, n = 7). RESULTS: The strength of the relationship between 8-OHdG and age was the same as that between pentosidine and age (the correlation coefficient between 8 OHdG and age was 0.33, pentosidine and age was 0.37). In addition, there was a positive and significant correlation between 8-OHdG and pentosidine. On the other hand, mean values of 8-OHdG and pentosidine showed no significant difference among the three groups. CONCLUSION: The results of the present study indicate that both 8-OHdG and pentosidine levels increase similarly in degenerative pathologic conditions. PMID- 11408025 TI - The activities of purine catabolizing enzymes in plasma and bronchial washing fluid in patients with lung cancer and pneumonia. PMID- 11408027 TI - Typical antipsychotics exhibit inverse agonist activity at rat dopamine D1-like receptors expressed in Sf9 cells. AB - The baculovirus system has been used to express the rat dopamine D1 receptors in Spodoptera frugiperda (Sf9) cells. A panel of typical antipsychotics including, alpha-flupenthixol, fluphenazine and thioridizine were found to inhibit dopamine dependent stimulation of adenylyl cyclase. However, these compounds were also found to inhibit adenylyl cyclase activity in the absence of agonist in Sf9 cells expressing dopamine D1-like receptors. Therefore, these nonselective dopamine receptor compounds displayed negative intrinsic or inverse agonist activity. None of the compounds tested were neutral antagonists. PMID- 11408028 TI - Capsaicin potentiates 1,25-dihydoxyvitamin D3- and all-trans retinoic acid induced differentiation of human promyelocytic leukemia HL-60 cells. AB - Human promyelocytic leukemia HL-60 cells are differentiated into monocytic or granulocytic lineage when treated with 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] or all-trans retinoic acid, respectively. In this study, the effect of capsaicin, an active component of the red pepper of the genus Capsocum, on cell differentiation was investigated in a HL-60 cell culture system. Treatment of HL-60 cells with 5 30 microg/ml capsaicin for 72 h inhibited cell proliferation and induced a small increase in cell differentiation. Interestingly, synergistic induction of HL-60 cell differentiation was observed when capsaicin was combined with either 5 nM 1,25-(OH)2D3 or 50 nM all-trans retinoic acid. Flow cytometric analysis indicated that combinations of 1,25-(OH)2D3 and capsaicin stimulated differentiation predominantly to monocytes whereas combinations of all-trans retinoic acid and capsaicin stimulated differentiation predominantly to granulocytes. Capsaicin enhanced protein kinase C activity in 1,25-(OH)2D3- and all-trans retinoic acid treated HL-60 cells. In addition, inhibitors for protein kinase C [bisindolylmaleimide (GF-109203X), chelerythrine, 1-(5-isoquinolinesulfonyl)-2 methylpiperazine dihydrochloride (H-7)] and an inhibitor for extracellular signal regulated kinase [2-(2'-amino-3'-methoxyphenyl)-oxanaphthalen-4-one (PD-098059)] significantly inhibited HL-60 cell differentiation induced by capsaicin in combination with either 1,25-(OH)2D3 or all-trans retinoic acid. These results indicate that capsaicin potentiates 1,25-(OH)2D3- or all-trans retinoic acid induced HL-60 cell differentiation and that both protein kinase C and extracellular signal-regulated kinase are involved in the cell differentiation synergistically enhanced by capsaicin. PMID- 11408029 TI - FK506 (tacrolimus) increases rat alpha1-acid glycoprotein expression in liver and primary cultured hepatocytes. AB - FK506 (tacrolimus) (10 mg/kg, s.c., 5 days) increased rat alpha1-acid glycoprotein (AGP) in serum and AGP mRNA in liver. FK506 potentiated the dexamethasone-increased AGP expression in primary cultured hepatocytes. In the luciferase promoter assay, FK506 potentiated the dexamethasone-increased promoter activity of the AGP gene in cultured rat hepatocytes, although FK506 alone had no effect on its activity. The combined effect of FK506 and dexamethasone was diminished by glucocorticoid responsive element (GRE) deletion and mutation or by an anti-glucocorticoid. These results indicated that FK506 causes the transcriptional induction of AGP, at least in part, via a glucocorticoid-mediated mechanism. PMID- 11408030 TI - Pharmacological blockade of brain alpha1-adrenoceptors as measured by ex vivo [3H]prazosin binding is correlated with behavioral immobility. AB - The present studies examined the relationship between the blockade of central alpha1-adrenoceptors, as measured by ex vivo binding of [3H]prazosin in the cerebral cortex and the inhibition of behavioral activation to a mildly novel environment (cage change). It was found that intraventricular (i.v.t.) terazosin, a saline-soluble alpha1-adrenoceptor antagonist, dose dependently inhibited both ex vivo cortical binding and behavioral activation and that there was a highly significant positive correlation between the two with a slope near unity. Prazosin, a nonsaline soluble antagonist which had to be given intraperitoneally (i.p.), was much less potent at blocking both behavioral activity and cortical ex vivo binding, although it blocked ex vivo binding in the lung, indicating that it was effective peripherally but did not readily enter the brain. Despite this, however, the inhibition of cortical binding and behavioral activation that i.p. prazosin did produce were highly correlated with each other and had a slope near unity as with terazosin, whereas the more potent inhibition of lung binding was less well correlated with behavioral inhibition and had a slope significantly less than one. These results confirm our earlier studies, which have shown that alpha1-adrenoceptor activity is essential for gross and fine motor behavior in the mouse and that prazosin, which is used extensively in behavioral research, has difficulty entering the mouse brain. PMID- 11408031 TI - 5-HT1A receptor activation and antidepressant-like effects: F 13714 has high efficacy and marked antidepressant potential. AB - To examine further the hypothesis that the magnitude of the intrinsic activity of agonists at 5-HT1A receptors determines the magnitude of their psychotropic activity, we studied the relationship between the maximal receptor activation produced by various 5-HT1A receptor ligands and their antidepressant-like effects (i.e., decreased immobility in the forced swimming test in rats). Using three different in vitro assays suitable to measure differences among high, intermediate, and low efficacy 5-HT1A receptor agonists, ligands were identified with intrinsic activities ranging from low-negative (i.e., the inverse agonist N [2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl)cyclohexane carboxamide (WAY 100635)) to high-positive (i.e., 3-chloro-4-fluorophenyl-(4 fluoro-4-[[(5-methyl-6-methylamino-pyridin-2-ylmethyl)-amino]-methyl]-piperidin-1 yl-methanone (F 13714)). In addition, novel compounds with intermediate intrinsic activity, like buspirone, but with high selectivity for 5-HT1A receptors, unlike buspirone, were identified. The maximal effects of the 5-HT1A receptor ligands in the forced swimming test correlated positively (rS=0.91, P<0.005) with the rank order of their intrinsic activity at 5-HT1A receptors. This relationship constitutes evidence that the magnitude of the psychotropic activity of 5-HT1A receptor ligands is a positive function of their intrinsic activity at the receptor, and suggests that F 13714, which had maximal effects in the forced swimming test significantly larger than any of the other compounds examined here, did so because of its higher intrinsic activity at 5-HT1A receptors. PMID- 11408032 TI - Reversal of neuroleptic-induced orofacial dyskinesia by 5-HT3 receptor antagonists. AB - Tardive dyskinesia, a syndrome of abnormal, involuntary hyperkinetic movements that occurs during long-term neuroleptic therapy is a major limitation of chronic neuroleptic therapy. The pathophysiology of tardive dyskinesia is still an enigma. The objective of the present study was to elucidate the role of 5-HT3 receptor involvement in neuroleptic-induced vacuous chewing movements in rats. Rats chronically (for 21 days) treated with haloperidol (1.5 mg/kg, i.p.) significantly developed vacuous chewing movements, as compared to vehicle-treated controls. Both ondansetron and tropisetron dose-dependently (0.25, 0.5 and 1.0 mg/kg, i.p.) reversed the haloperidol-induced vacuous chewing movements. Serotonin acting through 5-HT3 receptors might play a significant role in the pathophysiology of tardive dyskinesia, and 5-HT3 receptor ligands can be exploited as novel therapeutic agents for the treatment of tardive dyskinesia. PMID- 11408033 TI - Inhibitory effect of oxcarbazepine on high-frequency firing in peripheral nerve fibers. AB - We assessed the effects of oxcarbazepine, an antiepileptic derivative of carbamazepine, on discharges in single cutaneous afferent fibers produced by repetitive high-frequency stimulation (mimicking the abnormal excitation of peripheral nerves in neuropathic pain and paresthesia). After intravenous administration of oxcarbazepine, the later responses in the train dropped out without the earlier ones being affected and, thus, the total number of spikes decreased. The latency of the responses to an individual pulse was unchanged. These results, which indicate that oxcarbazepine inhibits the generation of high frequency firing without affecting impulse conduction, suggest that this drug may be useful against neuropathic pain and paresthesia. PMID- 11408034 TI - Hypolocomotor effects in rats of capsaicin and two long chain capsaicin homologues. AB - Capsaicin and its analogue N-arachidonoyl-vanillyl-amine (arvanil) are agonists of vanilloid VR1 receptors, and suppress spontaneous activity in mice through an unknown mechanism. Here, we tested in rats the effect on motor behavior of: (1) capsaicin; (2) N-linoleoyl-vanillyl-amine (livanil) and N-alpha-linolenoyl vanillyl-amine (linvanil), which, unlike arvanil, have very little affinity for cannabinoid CB1 receptors; and (3) the endocannabinoid anandamide (N-arachidonoyl ethanolamine), which is a full agonist at both cannabinoid CB1 and vanilloid VR1 receptors. All compounds, administered i.p., dose-dependently (0.1-10 mg/kg) inhibited ambulation and stereotypic behavior and increased inactivity in the open field test. The rank of potency observed in vivo (livanil>capsaicin>linvanil>anandamide) bore little resemblance with the relative potencies in a functional assay for rat vanilloid VR1 receptors (livanil=linvanil>capsaicin>anandamide) and even less with the relative affinities in rat CB1 receptor binding assays (anandamide>livanil>linvanil>capsaicin). The vanilloid VR1 receptor antagonist capsazepine (10 mg/kg, i.p.) blocked the effect of capsaicin but not of livanil or anandamide, whereas the CB1 receptor antagonist (N-(piperidin-1-yl)-5-(4 chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide.HCl (SR141716A, 3 mg/kg, i.p.) antagonized the actions of the CB1 receptor agonist Delta9-tetrahydrocannabinol, but not of livanil, anandamide or capsaicin. Anandamide occluded the effects of livanil on locomotion, possibly suggestive of a common mechanism of action for the two compounds. Finally, stimulation with capsaicin of cells expressing rat vanilloid VR1 receptors led to anandamide formation. These data suggest that motor behavior can be suppressed by the activation of: (1) vanilloid receptors, possibly via the intermediacy of anandamide; or (2) capsazepine- and SR141716A-insensitive sites of action for anandamide, livanil and linvanil, possibly the same that were previously suggested to mediate arvanil hypokinetic effects in mice. PMID- 11408035 TI - Chronic administration of losartan plus hydrochlorothiazide improves vascular status in young cardiomyopathic hamsters. AB - The combination of an angiotensin II receptor antagonist and a thiazide has been used extensively in the treatment of patients with overt heart failure. The effect of this combination on the vascular wall early in the disease, however, has not been investigated. To evaluate this effect, the vascular status of 3 month-old cardiomyopathic hamsters was assessed after daily administration of a combination of losartan (25 mg/kg, p.o.) and hydrochlorothiazide (6.5 mg/kg, p.o.) over an 8-week period. Age-matched golden hamsters were used as healthy controls. The contractile response of aortic rings to endothelin-1 was significantly higher in cardiomyopathic hamsters than in control animals. Concentration-response curves for the endothelin-1-induced contraction were displaced to the right after hydrochlorothiazide+losartan treatment (toward the curves for healthy controls); however, E(max) from treated hamsters was significantly reduced when compared to E(max) from untreated cardiomyopathic animals (1.016+/-0.073 vs. 1.346+/-0.153 g, P<0.05, n=6). No significant differences in the EC50 values from these curves were observed between hydrochlorothiazide+losartan treated and untreated cardiomyopathic animals (2.90+/-0.95 vs. 1.10+/-0.85 nM, P>0.05). The acetylcholine-induced relaxation observed in cardiomyopathic animals was not improved after treatment with hydrochlorothiazide+losartan or hydrochlorothiazide alone, but the combination of these drugs increased significantly the basal production of nitric oxide (NO). Angiotensin-converting enzyme activity increased in plasma (from 29.9+/-1.23 to 41.16+/-1.82 nmol x mg(-1) x min(-1), n=8, P<0.05) but decreased in the aorta (from 0.33+/-0.02 to 0.25+/-0.017 nmol x mg(-1) x min(-1), n=6, P<0.05) after treatment with hydrochlorothiazide+losartan. In addition, the combination of these drugs reduced the heart-to-body mass ratio (3.96+/-0.07 for treated vs. 5.01+/-0.20 mg/g for untreated animals, n=7, P<0.05), and the thickness of the aortic media (0.076+/-0.003 for treated vs. 0.149+/-0.009 mm for untreated animals, n=8, P<0.05). Although hydrochlorothiazide alone lowered systolic blood pressure to the same level achieved with both drugs in combination (from 166+/-10 for untreated cardiomyopathic animals to 84+/-1 mm Hg for hydrochlorothiazide+losartan, and 80+/-5 mm Hg for hydrochlorothiazide alone, P<0.05), no significant reduction in heart-to-body mass ratio was observed in animals treated with the diuretic alone (P>0.05). In conclusion, in this model of heart failure, chronic hydrochlorothiazide+losartan administration normalizes the vascular responses to endothelin-1, improves basal vascular tone, and prevents the development of cardiac and vascular hypertrophy. PMID- 11408036 TI - Monophosphoryl lipid A-induced delayed preconditioning is mediated by calcitonin gene-related peptide. AB - The delayed preconditioning of the heart by monophosphoryl lipid A is mediated by endogenous nitric oxide (NO), and the cardioprotection afforded by nitroglycerin is related to stimulation of calcitonin gene-related peptide (CGRP) release. The objective of this study was to explore whether improvement of preservation with cardioplegia by monophosphoryl lipid A is mediated by CGRP. In addition, we examined the effect of monophosphoryl lipid A on the tumor necrosis factor-alpha (TNF-alpha) content of myocardial tissues. The isolated rat heart was perfused in the Langendorff mode. Heart rate, coronary flow, left-ventricular pressure, and its first derivatives (+/-dp/dt(max)) were recorded, and plasma levels of NO and CGRP, the release of creatine kinase in coronary effluent and the content of TNF alpha in myocardial tissues were measured. Hypothermic ischemia for 4 h caused a decline in cardiac function, and an increase in the release of creatine kinase and in the content of TNF-alpha. Pretreatment with monophosphoryl lipid A (500 microg/kg, i.p.) for 24 h improved the recovery of cardiac function and reduced the release of creatine kinase concomitantly with a decrease in the content of cardiac TNF-alpha. Monophosphoryl lipid A markedly increased plasma concentrations of CGRP and NO. After pretreatment with L-nitroarginine methyl ester (L-NAME), the cardioprotection and the increased release of NO and CGRP induced by monophosphoryl lipid A were abolished. Capsaicin also abolished the cardioprotection and the increased release of CGRP induced by monophosphoryl lipid A, but did not affect the content of NO. The results suggest that monophosphoryl lipid A-induced preconditioning enhances preservation with cardioplegia and that the protective effects of monophosphoryl lipid A are related to stimulation of CGRP release. PMID- 11408037 TI - Factors that determine acetylcholine responsiveness of guinea pig tracheal tubes. AB - Acetylcholine administered to the inside of epithelium-denuded tracheal tubes did cause a potent contraction (2486+/-120 mg). In contrast, a response was hardly observed in tissues with an intact epithelial layer (674+/-81 mg), which was due to both the synthesis of nitric oxide and the activity of acetylcholinesterase, since the contractions to acetylcholine were significantly enhanced after preincubation with N(omega)-nitro-L-arginine methyl ester (L-NAME) or physostigmine (1374+/-65 and 1120+/-65 mg, respectively). In addition, the suppressive effect was caused by the barrier function of the epithelial layer, since preincubation of epithelium-denuded tissues with physostigmine significantly increased the pD2 value for acetylcholine (7.48+/-0.04) compared to intact tissues preincubated with physostigmine (6.32+/-0.10) and epithelium denuded preparations without physostigmine (6.37+/-0.06). Increasing concentrations of physostigmine administered to the inside of tissues with epithelium did induce a potent spontaneous contraction (1440+/-350 mg) that was prevented by atropine. In contrast to what was expected, the contractile response was diminished in tracheal tubes without epithelium (665+/-221 mg). It is concluded that contractions of epithelium-denuded tissues are more pronounced to exogenous than to endogenous acetylcholine, and that the production and breakdown of this neurotransmitter is very rapid in intact guinea pig airways. Moreover, the release of nitric oxide and the barrier function of the epithelium did suppress the responsiveness to acetylcholine. PMID- 11408038 TI - Stimulatory effects of nitric oxide donors on gastric acid secretion in isolated mouse stomach. AB - We previously reported on the stimulatory role of endogenous nitric oxide (NO) in gastric acid secretion. In the present study, we investigated the effects of NO donors on acid secretion in isolated mouse stomach. Nitroprusside (100 microM(-1) x mM) inhibited the gastric acid secretion induced by histamine (500 microM) in a concentration-dependent manner. In addition, nitroprusside abolished the acid secretion induced by bethanechol (100 microM) and by electrical stimulation (10 Hz) of the vagus nerve. On the other hand, nitroprusside, 75 microM, which did not affect the acid secretion induced by histamine, itself elicited an increase in acid secretion. The acid secretion induced by 75 microM nitroprusside was inhibited by 10 microM famotidine, a histamine H2 receptor antagonist. These results suggest that NO donors at high doses act on gastric parietal cells, resulting in inhibition of the stimulated acid secretion, and, at lower doses, facilitate histamine release from histamine-containing cells, leading to the increased acid secretion. PMID- 11408039 TI - Effects of cyclosporin A administered into the airways against antigen-induced airway inflammation and hyperreactivity in the rat. AB - The immunosuppressant cyclosporin A given orally has anti-asthma properties but carries an undesirable risk of systemic effects. We administered cyclosporin A to Brown Norway rats either orally (p.o.) or topically by intratracheal (i.t.) instillation into the airways before inhaled antigen. Cyclosporin A suppressed the antigen-induced accumulation of activated (CD25+) CD4+ T lymphocytes and eosinophils in the lung, interleukin-5 mRNA expression in lung tissue and airway hyperreactivity. Intratracheal cyclosporin A suppressed cell accumulation at a 10 fold lower dose than that required orally. Minimum effective doses were 3 mg x kg(-1) i.t. and 30 mg x kg(-1) p.o. Intratracheal administration reduced the plasma concentration and systemic exposure compared with an equieffective oral dose, but the reduction (4-5-fold) was not as large as anticipated. Our data suggests that although topical administration to asthmatics would provide some potential for an improved safety margin, it may not offer any major advantage over existing oral therapy. However, the data clearly demonstrate that a novel immunosuppressant with similar anti-inflammatory properties but reduced potential for systemic effects would offer an attractive therapy for severe asthma. PMID- 11408040 TI - Helicobacter pylori lipopolysaccharide-provoked injury to rat gastroduodenal microvasculature involves inducible nitric oxide synthase. AB - The actions of a purified Helicobacter pylori lipopolysaccharide (3 mg x kg(-1), i.v.) on rat gastric antral and duodenal microvascular integrity (determined as radiolabelled albumin leakage) and the expression of the inducible nitric oxide (NO) synthase (iNOS; assessed by the citrulline assay) were investigated 4 h after challenge. Significant increases of albumin leakage and expression of iNOS in both antral and duodenal tissues were observed following challenge. Concurrent administration of the selective iNOS inhibitor, 1400W (N-(8-(aminomethyl)benzyl) acetamidine; 0.2-1 mg x kg(-1), s.c.), with lipopolysaccharide, caused a dose dependent attenuation of the gastric and duodenal albumin leakage. Thus, H. pylori lipopolysaccharide can initiate the expression of iNOS in the stomach and duodenum following systemic challenge, which can provoke gastroduodenal microvascular dysfunction. PMID- 11408041 TI - Fluoxetine increases the content of neurotrophic protein S100beta in the rat hippocampus. AB - Recent studies indicate that a protracted daily administration of the antidepressant fluoxetine to adult rats increases cell proliferation/neurogenesis in the hippocampus. It has been hypothesized that this action of fluoxetine might be mediated by neurotrophic factors. We hypothesized that glial S100beta could be such a factor, and using quantitative Western immunoblotting, we investigated the effect of a 21-day treatment of rats with fluoxetine (5 mg/kg), and found that fluoxetine increases the content of hippocampal S100beta. PMID- 11408042 TI - Accelerated functional recovery after neuronal injury by P2 receptor blockade. AB - The effect of the P2 receptor antagonist pyridoxalphosphate-6-azophenyl-2',4' disulphonic acid (PPADS) on changes of the quantitative electroencephalogram (EEG) after injury of rat brain tissue was investigated. PPADS accelerated the functional recovery from microdialysis probe-induced disturbances in the nucleus accumbens by a decrease of the ratio of absolute slow (0.6-4 Hz) to fast (8-30 Hz) power, mainly caused by a decrease in the delta frequency power. These data provide evidence for a possible neuroprotective effect of P2 receptor antagonists in vivo. PMID- 11408043 TI - Kinetics of glutathione and daunorubicin efflux from multidrug resistance protein overexpressing small-cell lung cancer cells. AB - The present study examined how the multidrug resistance protein (MRP1), which is an ATP-dependent anionic conjugate transporter, also mediates the transport of reduced glutathione (GSH) and the co-transport of the cationic drug, daunorubicin, with GSH in living GLC4/Adr cells. To obtain information on the affinity of GSH for the multidrug resistance protein in GLC4/Adr cells, we investigated the GSH concentration dependence of the ATP-dependent GSH efflux. The intracellular GSH concentration was modulated by preincubation of the cells with 25 microM buthionine sulfoximine, an inhibitor of GSH synthetase, for 0-24 h. The transport of GSH was related to the intracellular GSH concentration up to approximately 5 mM and then plateaued. Fitting of the obtained data according to the Michaelis-Menten equation revealed a Km of 3.4+/-1.4 mM and a Vmax of 1.5+/ 0.2x10(-18) mol/cell/s. The ATP-dependent transport of GSH was inhibited by 3 ([[3-(2-[7-chloro-2-quinolinyl]ethenyl)phenyl]-[(3-dimethylamino-3-oxopropyl) thio]-methyl]thio)propanoic acid (MK571), with 50% inhibition being obtained with 1.4 microM MK571. We investigated the GSH concentration dependence of the MRP1 mediated ATP-dependent transport of daunorubicin under conditions where the transport of daunorubicin became saturated. The daunorubicin transport was related to the intracellular GSH concentration up to approximately 5 mM and then plateaued. We were therefore in the situation where GSH acted as an activator: its presence was necessary for the binding and transport of daunorubicin by MRP1. However, GSH was also transported by the multidrug resistance protein. The concentration of GSH that gave half the maximal rate of daunorubicin efflux was 2.1+/-0.8 mM, very similar to the Km value obtained for GSH. In conclusion, the rate of daunorubicin efflux, under conditions where the transport of daunorubicin became saturated, and the rate of GSH efflux determined at any intracellular concentration of GSH were very similar, yielding a 1:1 stoichiometry with respect to GSH and daunorubicin transport. These results support a model in which daunorubicin is co-transported with GSH. PMID- 11408044 TI - Nature of the oligomers formed by muscarinic m2 acetylcholine receptors in Sf9 cells. AB - Wild-type, FLAG-tagged, and c-myc-tagged muscarinic m2 receptors extracted in digitonin-cholate from singly and co-infected Sf9 (Spodoptera frugiperda) cells were indistinguishable in their binding of [3H]quinuclidinylbenzilate, either before or after purification. The FLAG epitope was found to coimmunoprecipitate with the c-myc epitope when co-infected cells were solubilised in digitonin cholate, n-dodecyl-beta-D-maltoside or Lubrol-PX. The degree of coprecipitation in digitonin-cholate was unaffected by preincubation of the extract for up to 60 min at 30 degrees C, with or without muscarinic receptor ligands; no coimmunoprecipitation occurred in mixed extracts from singly infected cells. As measured by [3H]quinuclidinylbenzilate, the efficiency of immunoprecipitation from co-infected cells was 87% of that from singly infected cells. The amount of receptor immunoprecipitated from the latter, as determined by densitometry, was 2.3-fold that expected from the loss of binding from the extract. The data suggest that at least some of the receptors were trimeric or larger and that oligomers neither formed nor dissociated under the conditions of the experiments. Also, some receptors appear to be non-functional or latent in digitonin solubilised extracts. PMID- 11408045 TI - Negative functional effects of cGMP mediated by cGMP protein kinase are reduced in T4 cardiac myocytes. AB - We tested the hypothesis that in isolated rabbit cardiac myocytes, the negative functional effects of cyclic GMP are partly mediated by cyclic GMP-dependent protein kinase activity, and that these effects are altered in thyroxine (T4, 0.5 mg/kg/day for 16 days)-induced hypertrophic myocytes. Using isolated ventricular myocytes from control (N=8) and T4 (N=8) hypertrophic hearts, data for percent cell shortening (%) and maximum rate of contraction (microm/s) were collected using a video edge detector at baseline, after the addition of 10(-6) M 8-bromo cyclic GMP (8-Br-cGMP), 10(-5) M 8-Br-cGMP, and 10(-6) M KT5823 (10-methoxy-10 methoxycarbonyl-9, 10, 11, 12-tetrahydro-9, 12-epoxy-(1H)-diinidolo [1, 2, 3, f g: 3', 2', 1'-k-j]-pyrrolidino-[3,4-i] [1,6]-benzodiazocin-2-methyl-1-one, cyclic GMP protein kinase inhibitor). Protein phosphorylation was determined autoradiographically after gel electrophoresis. In both control and T(4) myocytes, 8-Br-cGMP caused a significant decrease in percent shortening (5.56+/ 0.49% to 3.02+/-0.47% in control and 4.34+/-0.33% to 3.13+/-0.17% in T4 myocytes) and maximal rate of contraction 57.35+/-6.05 to 36.82+/-3.17 microm/s in control and 58.49+/-3.28 to 42.88+/-2.29 microm/s in T4 myocytes). KT5823 significantly increased percent shortening to 3.77+/-0.28% and rate to 48.68+/-4.71 microm/s after 8-Br-cGMP only in control myocytes. In T4 myocytes, the changes in percent shortening and rate after KT5823 were not significant. Protein phosphorylation was increased by 8-Br-cGMP in control and to a lesser extent in T4 myocytes, but the increment was reduced by KT-5823 in control only. These data demonstrated that cyclic GMP had negative functional effects partially mediated by cyclic GMP protein kinase in control myocytes. Cyclic GMP also exerted negative functional effects in thyroxine-induced hypertrophic myocytes, but cyclic GMP protein kinase activity was not an important regulator of these effects in T4 ventricular myocytes. PMID- 11408046 TI - Increased behavioral neurosteroid sensitivity in a rat line selectively bred for high alcohol sensitivity. AB - Acute administration of a neurosteroid 5beta-pregnan-3alpha-ol-20-one induced a greater impairment in motor performance of the selectively bred alcohol-sensitive (ANT) than alcohol-insensitive (AT) rats. This difference was not associated with the sensitivity of gamma-aminobutyrate type A (GABA(A)) receptors, as 5alpha pregnan-3alpha-ol-20-one (allopregnanolone) decreased the autoradiographic signals of t-butylbicyclophosphoro[35S]thionate binding to GABA(A) receptor associated ionophores more in the brain sections of AT than ANT rats. Nor was the difference associated with baseline levels of neuroactive progesterone metabolites, as 5alpha-pregnan-3,20-dione (5alpha-DHP) and 5alpha-pregnan-3alpha ol-20-one were lower in the ANT rats. After ethanol (2 g/kg, i.p.) administration and the subsequent motor performance test, the increased brain concentrations of these metabolites were still lower in the ANT than AT rats, although especially in the cerebellum the relative increases were greater in the ANT than AT rats. The present data suggest that the mechanisms mediating neurosteroid-induced motor impairment are susceptible to genetic variation in rat lines selected for differences in ethanol intoxication. PMID- 11408047 TI - Antisera against endogenous opioids increase the nocifensive response to formalin: demonstration of inhibitory beta-endorphinergic control. AB - The roles of endogenous opioid peptides in the brain in the modulation of nocifensive responses to formalin in ICR mice were studied. Mice were pretreated intracerebroventricularly (i.c.v.) with rabbit antiserum against beta-endorphin, [Leu5]enkephalin, [Met5]enkephalin or dynorphin A-(1-17) 1 h prior to intraplantar injection of formalin (0.5%, 25 microl) and the nocifensive licking responses were then observed. Pretreatment of mice with antiserum against beta endorphin enhanced the second phase, but not the first phase of the nocifensive responses to formalin. Pretreatment with antiserum against [Leu5]enkephalin also caused a small but statistically significant enhancement of the second phase, but not the first phase of nocifensive responses to formalin. On the other hand, pretreatment with antiserum against [Met5]enkephalin or dynorphin A-(1-17) did not affect the nocifensive response to formalin. Our results indicate that beta endorphinergic, and to a lesser extent, [Leu5]enkephalinergic systems are activated at the supraspinal sites to attenuate the nocifensive responses to formalin stimulation. PMID- 11408048 TI - Hypoxia impairs endothelium-dependent relaxation in organ cultured pulmonary artery. AB - In intrapulmonary arteries cultured under hypoxic conditions (5% oxygen) for 7 days, endothelium-dependent relaxation and cGMP accumulation induced by substance P were decreased as compared to those of a normoxic control (20% oxygen). In rabbit mesenteric arteries exposed to chronic hypoxia, however, endothelial dysfunction was not observed. Furthermore, in endothelium-denuded pulmonary arteries exposed to hypoxia, neither relaxation nor cGMP accumulation due to sodium nitroprusside differed from those of the normoxic control. Hypoxia did not change the mRNA expression of endothelial NO synthase (eNOS), the protein expression of eNOS or the eNOS regulatory protein caveolin-1 as assessed by semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) or whole-mount immunostaining. Morphological study revealed atrophy of endothelial cells and condensation of the eNOS protein in many cells. These results suggest that chronic hypoxia impaired NO-mediated arterial relaxation without changing either the eNOS protein expression or the NO-sensitivity of smooth muscle cells in pulmonary arteries. Changes in cell structure and organization may be involved in endothelial dysfunction. PMID- 11408049 TI - Effect of sodium tauroursodeoxycholate on phalloidin-induced cholestasis in rats. AB - We investigated the therapeutic effect of tauroursodeoxycholate on phalloidin induced cholestasis in rats. Intrahepatic cholestasis was induced by administration of phalloidin (500 microg/kg, i.p.) for 7 days. From the day of the last phalloidin injection, tauroursodeoxycholate (60-360 micromol/kg) was given intravenously twice a day for 4 days. On the next day after the last tauroursodeoxycholate administration, bile flow, serum biochemical parameters and biliary lipid excretion rates were determined. Tauroursodeoxycholate significantly suppressed the decrease in bile flow and increases in serum alkaline phosphatase, leucine aminopeptidase and glutamic pyruvic transaminase activities, cholesterol, phospholipid and bile acid concentrations observed in phalloidin-induced cholestasis in rats. Furthermore, tauroursodeoxycholate significantly improved the biliary cholesterol and phospholipid excretion rates in phalloidin-induced cholestasis in rats. These results demonstrate the usefulness of tauroursodeoxycholate as a therapeutic agent in intrahepatic cholestasis. PMID- 11408050 TI - Receptor constants for endomorphin-1 and endomorphin-1-ol indicate differences in efficacy and receptor occupancy. AB - The opioid properties of endomorphin derivatives containing a C-terminal alcoholic(-ol) function were compared to the parent amidated compounds in isolated organs (longitudinal muscle strip of guinea-pig ileum and mouse vas deferens). Similar data were also generated for the mu-opioid receptor selective agonist synthetic peptide (D-Ala2, MePhe4, Gly5-ol)-enkephalin (DAMGO) and its Gly5-NH2 congener (DAMGA). Endomorphin-1-ol (Tyr-Pro-Trp-Phe-ol) had an IC50 of 80.6 nM in mouse vas deferens and 61.2 nM in guinea-pig ileum; the corresponding values for endomorphin-2-ol (Tyr-Pro-Phe-Phe-ol) were 49.6 and 48.2 nM, for DAMGO 59.8 and 29.2 nM, respectively. As it was indicated by the antagonism by naltrexone, the agonist actions were exerted exclusively at mu-opioid receptors in both organs. The -ol derivatives were slightly (2.3-4.3 times) less potent than the parent amides in the bioassays: all peptides had, apparently, full agonist properties in intact preparations. With the aim of revealing potential partial agonist properties among the investigated peptides, we partially inactivated the mu-opioid receptor pool in mouse vas deferens by 5x10(-7) M beta funaltrexamine. The calculated receptor constants indicated a "high-affinity, low intrinsic efficacy" profile (i.e. a potential partial agonist property) for endomorphin-1, an intermediate character for endomorpin-1-ol and full agonism for DAMGA and DAMGO. Apparently, a higher receptor fraction remained accessible for endomorphin-1 (42.8%) than for the -ol congener (14.0%), DAMGO (20.2%) and DAMGA (14.1%) after partial inactivation. PMID- 11408051 TI - Are emotions contagious? Evoked emotions while viewing emotionally expressive faces: quality, quantity, time course and gender differences. AB - In human interactions, frequently one individual becomes 'infected' with emotions displayed by his or her partner. We tested the predictions by Hatfield et al. (1992) (Primitive emotional contagion. Review of Personal and Social Psychology 14, 151-177) that the automatic, mostly unconscious component of this process, called 'primitive emotional contagion', is repeatable and fast, that stronger facial expressions of the sender evoke stronger emotions in the viewer and that women are more susceptible to emotional contagion than men. We presented photos from the Pictures of Facial Affect (Ekman and Friesen, 1976). (Pictures of Facial Affect. Consulting Psychologists Press, Palo Alto) on a PC varying the affective content (happy and sad), the expressive strength and the duration of presentation. After each photo, subjects rated the strength of experienced happiness, sadness, anger, disgust, surprise, fear and pleasure. Feelings of happiness or sadness were significantly, specifically and repeatedly evoked in the viewer - even with presentations lasting only 500 ms. Stronger expressions evoked more emotion. The gender of the viewer had weak effects. We hypothesize that this fast and repeatable reaction is likely to have a 'prewired' neural basis. We propose that the induction of emotional processes within a subject by the perception of emotionally expressive faces is a powerful instrument in the detection of emotional states in others and as the basis for one's own reactions. Detailed knowledge of emotional reactions to faces is also valuable as a basis for psychiatric studies of disorders in affect and/or communication and in studies using functional imaging (fMRI or PET) where faces are increasingly used as stimuli. PMID- 11408052 TI - Impact of threat relevance on P3 event-related potentials in combat-related post traumatic stress disorder. AB - The purpose of this study was to examine electrophysiological response to trauma relevant stimuli in combat-related post-traumatic stress disorder (PTSD). Study design incorporated comparison of 10 Vietnam War veterans with PTSD diagnosis to 10 Vietnam War veterans with no mental disorder diagnosis on P3 components in a series of two oddball tasks (trauma-relevant threat, trauma-irrelevant threat) counterbalanced for order. Each task included high probability emotionally neutral distractor words and low probability neutral target words, but differed in the content of low probability threat words. Whereas threat words in the trauma-relevant oddball task pertained directly to combat trauma, threat words in the trauma-irrelevant oddball task were socially threatening words. Results revealed that, in comparison to healthy combat veterans, those diagnosed with PTSD demonstrated: (a) attenuated P3 response to neutral target items at selected electrode sites across both oddball tasks; and (b) increased responsivity to trauma-relevant combat stimuli but not to trauma-irrelevant social-threat stimuli at frontal electrode sites (F3, F4). Results are consistent with resource allocation models of PTSD, which suggest that PTSD is characterized by attentional bias to threat stimuli at the expense of attention to emotionally neutral information. PMID- 11408053 TI - Dual-task performance in depressed geriatric patients. AB - Older patients suffering from a major depression are often impaired on tasks that require executive control processes. However, a wide variety of executive abilities exist in humans, and it is not clear that all are impaired in depression or that such impairments persist beyond remission of the depression. One executive process that plays a central role in mental operations such as working memory is the ability to co-ordinate the simultaneous performance of multiple tasks. Dual-task performance has been extensively studied in normal subjects but there is little work with depressed patients. The present study examined the performance of depressed (M age=71.0, S.D.=7.1) and control subjects (M age=69.3, S.D.=7.0) on two tasks (visual tracking and backward digit span), both when each task was carried out by itself and when the two tasks were carried out simultaneously. Dual-task performance was impaired in depressed patients prior to antidepressant treatment and this impairment persisted even after remission of the depression. These results suggest that, like other executive abilities, the ability to schedule and co-ordinate the conflicting processing demands present in a dual-task situation is impaired in depressed geriatric patients and that this impairment may be a trait effect. PMID- 11408054 TI - Correlation between serum lipid concentrations and psychological distress. AB - This study examines the correlations between serum lipid levels and psychological distress. There were 4444 consecutive attendees of general health clinics who participated in the study. Psychological symptoms were measured by the Taiwanese version of the Symptoms Check List 90, revised (T-SCL-90-R). Levels of fasting serum lipids, including total cholesterol, total triglycerides and high-density lipoprotein cholesterol (HDL-C), were determined. Multiple linear regression analyses, with adjustment for confounders, revealed that the concentration of HDL C had significant inverse associations with scores of depression, somatization and phobic anxiety. Women with an HDL-C level lower than 35 mg/dl scored significantly higher on depression, interpersonal sensitivity, phobia, anxiety, somatization and aggressive hostility, while subjects with a total cholesterol concentration lower than 160 mg/dl scored significantly higher on anxiety, aggressive hostility, phobia, and psychoticism. This study provides, for the first time, comprehensive data derived from the Taiwanese population on the link between lipids and psychological symptoms, revealing a reverse correlation between depression and serum concentrations of HDL-C. PMID- 11408055 TI - Assessment of reliability in the clinical evaluation of depressive symptoms among multiple investigators in a multicenter clinical trial. AB - The objective of this work was to determine the severity of depressive symptoms when multiple clinical examiners evaluate a single subject, as preparatory to their participation as evaluators in a clinical trial. Using the 17-item Hamilton Depression Rating Scale (HDRS), 37 psychiatrists independently assessed the videotape of a patient with symptoms of depression. A new measure for the detection of multiple examiners not in consensus (DOMENIC) was used to identify scale items with low reliability and raters with low inter-rater reliability, from among the remaining raters. Overall inter-rater agreement on the full HDRS was 'excellent' (97%). All raters but one showed adequate agreement both on individual items and on total scores. Two of the 17 HDRS symptomatology items had unacceptable levels of inter-rater scoring variability (<70% agreement). The use of DOMENIC allows for the detection of items of low inter-rater reliability and identification of raters that deviate from the group's ratings prior to the beginning of a clinical trial. PMID- 11408056 TI - Use of structural equation modeling to test the construct validity of a case definition of Gulf War syndrome: invariance over developmental and validation samples, service branches and publicity. AB - To attempt to replicate the syndrome-like structure identified by exploratory factor analysis of symptom reports from 249 Gulf War veterans of a Naval reserve battalion (the developmental sample), we administered Haley's original symptom questionnaire to 335 Gulf War veterans who served primarily in active-duty US Army units living in North Texas (the validation sample). On the basis of recently validated goodness-of-fit criteria (SRMRor=0.95), a structural equation model (Model 1) with four symptom scales loading on each of three first-order latent syndrome factors fit both the developmental and validation samples well and was invariant across both samples. Additional models validated a higher-order latent factor (a single Gulf War syndrome) explaining the variances and covariances of the first-order factors, four additional symptom scales loading on the higher-order factor, and four possible secondary factor loadings that also fit the data well. All structural models were invariant across cohorts of the validation sample surveyed before and after intense publicity following publication of the case definition. These findings suggest that the apparent syndrome structure of a single Gulf War syndrome with three variants may be found widely and justify a confirmatory sample survey of Gulf War-era veterans. PMID- 11408057 TI - Ossifying fibromyxoid tumor of soft parts: report of a case with novel cytogenetic findings. AB - A slowly growing tumor of the left thenar region in a 40-year-old man had the classic features of an ossifying fibromyxoid tumor of soft parts, including an incomplete shell of lamellar bone; a center composed of nodular aggregates of small spindled, oval, and stellate cells in abundant myxoid stroma; and strong expression of vimentin, S-100, and neuron-specific enolase by the tumor cells. Clonal chromosomal abnormalities included loss of a chromosome 6, extra material of unknown origin attached to the long arm of chromosome 12, and an unbalanced translocation involving the short arm of a chromosome 6 and the long arm of a chromosome 14. The karyotype was interpreted as 45,XY, der(6;14)(p10;q10),add(12)(q24.3). The chromosomal abnormalities suggest osteochondroblastic rather than neuronal or schwannian lineage. PMID- 11408058 TI - Novel cell lines established from a human myxoid malignant fibrous histiocytoma arising in the uterus. AB - Two cell lines (Nara-H and Nara-F) with different phenotypes were established from a myxoid MFH of the uterus. In vitro, Nara-F grew in sheets showing a storiform arrangement and Nara-H in raised colonies. Although tumors generated in nude mice shared similar morphological features of abundant myxoid tumor in Nara H and -F, the pleomorphic component was conspicuous in Nara-F. Both cell lines produced hyaluronic-acid but CD44 was expressed only in Nara-H. Estrogen receptor alpha (ER alpha) and progesterone receptor (PgR) were detected in Nara-H. Nara-F was positive for ER beta and PgR. Among hormonal agents, the response to the anti estrogen tamoxifen was more sensitive than progesterone agents. This report illustrates the characteristics of these newly established cell lines, and presents the possibility of an adjuvant hormonal therapy for MFH. PMID- 11408059 TI - Frequent co-amplification of two different regions on 17q in aneuploid breast carcinomas. AB - Chromosome 17q is highly susceptible to rearrangement mutations in breast cancer. c-erbB-2 at 17q11.2 approximately q21.1 is frequently amplified, as is a region at 17q22 approximately q24. As a step in the search for the target gene(s) of the 17q22-q24 amplification we determined whether the placental lactogen (PL) genes at 17q23 were amplified in 59 breast carcinomas. These genes were selected as their upregulation could theoretically be involved in breast cancer tumorigenesis. Amplification of the PL genes, and also of c-erbB-2, was detected using semi-quantitative PCR. The reliability of this method was confirmed since c erbB-2 results obtained using PCR, Southern blotting and immunohistochemistry were in good agreement. The PL genes were amplified in 13 (22%) of the tumors. Furthermore, the PL and c-erbB-2 genes were frequently co-amplified although there is a non-amplified region between them. Expression of PL was investigated in 26 tumors and was detected in 16 of these cases including all 10 tumors with amplification of the PL genes. The tumors with PL gene amplification were all aneuploid. A trend was seen towards an increased incidence of lymph node involvement for tumors with amplification of the PL genes and for tumors with co amplification of PL and c-erbB-2, which suggests a possible association with high malignancy. PMID- 11408060 TI - Comparative genomic hybridization analysis reveals 3q gain resulting in genetic alteration in 3q in advanced oral squamous cell carcinoma. AB - We analyzed DNA sequence copy number aberrations (DSCNAs) in 17 primary oral squamous cell carcinomas (OSCCs) by comparative genomic hybridization. DSCNAs were detected frequently at 3q25-qter (7/17), Xp21 (5/17), and Xq12-q23 and 8q23 q24 (4/17), and losses were detected frequently at 13q21-q22 (5/17), 3p21-pter, 4p15-pter and 17p13 (4/17), and 8p22-pter and 9p21-pter (3/17). Four tumors showed amplifications of seven loci: 3q11-qter, 3q13, 3q26, 7q21-q22, 8q23-qter, 9p22-pter, and 12p11. The total number of DSCNAs was significantly greater in stage III and stage IV tumors than in stage I and stage II tumors (P=.008). Furthermore, 3q gain was detected preferentially in stage III and stage IV tumors (6/8) rather than in stage I and stage II tumors (1/9, P=.013). In our study, all tumors with gain of 3q also contained one or more loss(es) in common regions. On the other hand, all tumors with gain of 9p did not contain 3q gains. These observations indicate that gain of 3q and accumulation of DSCNAs are strongly associated with tumor progression in OSCC. Furthermore, 3q gain and loss of one or more additional loci in common aberration regions appears to be a group of DSCNs associated with dominant genetic pathways of leading to advanced OSCCs. PMID- 11408061 TI - Consistent DNA losses on the short arm of chromosome 1 in a series of malignant gastrointestinal stromal tumors. AB - Fourteen malignant gastrointestinal stromal tumors (GISTs), characterized by immunohistochemistry, were analyzed by comparative genomic hybridization (CGH). The most striking feature was the detection of consistent DNA losses on the short arm of chromosome 1 in these 14 malignant tumors. Additional recurrent imbalances were also found: significant gains, which could be indicative of tumor progression, were frequent on the long arm of chromosome 1, as were losses of DNA copy number detected in chromosomes 13, 14, 15 and 22. PMID- 11408062 TI - Analysis of genetic alterations in salivary gland tumors by comparative genomic hybridization. AB - In order to define and map chromosomal copy number alterations in salivary gland tumors (SGTs), a comparative genomic hybridization (CGH) technique was applied to two pleomorphic adenomas (PAs), one adenoid cystic carcinoma (ACC), and one basal cell adenocarcinoma (BCAC). The PAs exhibited regional copy number losses at 5q12.4-q14.1, 9q12-q21.13, and 16q11.2, as well as a gain at 20p12.1; among these, the losses at the 9q12-q21.11 and 16q11.2 regions were common to both PAs. The ACC showed overrepresentations of the entire regions of chromosomes 16 and 20, a regional gain at 22q12.3-q13.1, and no losses. In the BCAC, regional gains at 9p21.1-pter, 18q21.1-q22.3, and 22q11.23-q13.31 as well as losses at 2q24.2 and 4q25-q27 were seen; the gain at 22q12.3-q13.1 was common in both the ACC and the BCAC. These CGH data indicate that different genetic alterations are present in the different types of SGTs, and that the alterations involve several chromosomes. The discovery of common alterations in the same and/or different types of tumors might be important in the understanding of the development and progression of the SGTs. PMID- 11408063 TI - Triplication of 1q in Fanconi anemia. AB - We report herein a 38-year-old male patient with Fanconi anemia but with few phenotypic manifestations--short stature, sterility, and hypoplasic anemia with several years of evolution-who developed a myelodysplastic syndrome (MDS). Bone marrow karyotype showed long arm triplication of chromosome 1 (q12-21q31-q32), and two markers add(11)(p15) and add(21)(q22) which had extra material of chromosome 3 besides the normal chromosome 3 pair. Peripheral blood showed chromosome instability; SCE was normal. Both the patient and his family showed a high prevalence of malignant diseases. 1q duplication and, in a few cases, triplication of 1q has been related to Fanconi anemia, being of unknown significance. PMID- 11408064 TI - Cytogenetic characterization of Ewing tumors using fine needle aspiration samples. a 10-year experience and review of the literature. AB - Chromosomal analysis was performed in fine needle aspiration samples of 98 primary Ewing tumors (ETs) prior to treatment. Among the 58 (59.18%) successful cultures, t(11;22)(q24;q12) was observed in 87.9% and 6.8% had abnormalities other than t(11;22), viz., del(22)(q12), der(16)t(1;16)(q12;q11), and variant t(8;22)(q24;q12). Involvement of breakpoints 1q21, 1q22, 3p14, 16q22, and 17p13 was also observed. Numerical abnormalities such as trisomies 8 and 12 were found in 29.3% and 20.6% and trisomy 18 in 17.2%. An attempt was made to evaluate the role of these additional changes in the process of tumor development, metastasis, and progression of the disease. This is the largest cytogenetic study on ET from a single center using a simple and reliable technique of fine-needle aspiration culture. The literature on cytogenetics of ET is reviewed. PMID- 11408065 TI - Pattern of chromosomal imbalances in non-B virus related hepatocellular carcinoma detected by comparative genomic hybridization. AB - Only limited data are available on comparative genomic hybridization (CGH) in hepatocellular carcinoma (HCC). They concern mainly B virus related HCC. Therefore, we used CGH to detect chromosomal imbalances in 16 non-B virus related HCC in alcoholic cirrhosis in 7 cases (HA1 to HA7), in C virus cirrhosis in 7 cases (HC1 to HC7), in non-cirrhotic liver in 2 cases (NC1, NC2), and in 9 non malignant cirrhotic tissues. The most frequent imbalances in HCC were gains of whole chromosomes or chromosomal regions 7 or 7q (10/16, 62%), 1q (9/16, 56%), 5 or 5q (9/16, 56%), 8q (8/16, 50%), 6p (6/16, 37%), 15q (5/16, 31%), 20 or 20q (5/16, 31%), and losses of 17p (6/16, 37%), and 8p (5/16, 31%). High-level gains were identified in HCC on 1q (2/16), 3q (1/16), 7q (1/16), and 8q (3/16). No chromosomal imbalances were detected in any of the cirrhotic tissues. Most of the gains, losses, and amplifications detected in this CGH study corresponded well to those identified in previous studies, except for gains of whole chromosome 5 or 7 and/or of chromosome arms 5q or 7q and losses on 4q. Our results suggest that other chromosomal regions are involved in hepatocarcinogenesis. PMID- 11408066 TI - DNA copy number profiling in esophageal Barrett adenocarcinoma: comparison with gastric adenocarcinoma and esophageal squamous cell carcinoma. AB - We screened 18 specimens of Barrett adenocarcinoma for genetic alterations using comparative genomic hybridization (CGH) to analyze DNA copy number changes. The most common gains were at 20q (56%) and 17q (39%). High-level amplifications were observed in the same chromosomes. The most common losses were in chromosomes 4 (22%) and 5 (22%). Other recurrent changes were gains of chromosomes 8, 10q, and 13. We compared the copy number changes in Barrett adenocarcinoma and those previously reported in the intestinal type of stomach carcinoma. The similarities we found suggest a common molecular pathogenesis, whereas dissimilarities seen between Barrett adenocarcinoma and esophageal squamous cell carcinoma are in keeping with a well-known different etiology. PMID- 11408067 TI - Comparative genomic hybridization analysis of transitional cell carcinomas of the renal pelvis. AB - We used comparative genomic hybridization to analyze 10 primary tumor samples from patients with transitional cell carcinoma of the renal pelvis. The most frequent loss was located at 9q, that is, in 50% of the tumors. Gains of DNA sequences were most frequently observed in chromosome regions 1q21 approximately q23, 2p23 approximately p25, 8q21.1 approximately q22 and in the whole chromosome 20. High level amplifications at 1q21 approximately q25, 6p22 approximately p23, 8q21 approximately q22, 8q22 approximately q24.1, 11q13, and 12q14 approximately q21 were detected. Most of these regions have previously been reported to be involved in transitional cell carcinoma of the bladder, thus confirming the importance of an increasing number of chromosome imbalances in the development and progression of this type of tumors. PMID- 11408068 TI - Evaluation of loss of heterozygosity/allelic imbalance scoring in tumor DNA. AB - Loss of heterozygosity and allelic imbalance in tumors are usually detected by either radioactive labeling of PCR products with subsequent scoring of autoradiographs or by a semi-quantitative fluorescence-based protocol. Polymorphic microsatellite loci are the most common marker type used in these studies. Even though no consensus exists as to how to evaluate such data, results are often compared directly between studies applying the two different protocols. In the present study, we analyzed twice by each protocol three loci in 60 blood/tumor pairs, finding good correlation between the results obtained by the two methods. However, a higher sensitivity and the possibility to correct for stutter peaks were among several advantages inherent in the fluorescence labeling approach. In addition, we determined the cut-off level for allelic imbalance scoring by the fluorescent primer protocol, by repeated analysis of 485 constitutional heterozygous genotypes at 20 different dinucleotide repeat loci. Based on the standard deviation, we found that allelic imbalance should be scored whenever the peak height of one allele in tumor DNA is reduced to less than 0.84 of its value in constitutional DNA, relative to the other allele. Applying this cut-off value, more imbalances are detected than by the visual scoring of autoradiographs. Our data therefore suggest that a lower threshold value (0.75) must be used when results from both fluorescent and radioactive assays are compared. PMID- 11408069 TI - Acute myeloid leukemia with t(5;18)(q35;q21). AB - A case of acute myelocytic leukemia with a translocation (5;18)(q35;q21) is reported. Cytogenetic abnormalities of the long arm of chromosome 5 have long been known to affect hematopoiesis. Although translocations between 5q and other chromosomes have been associated with malignancy, this is the first reported case of a t(5;18) resulting in acute myeloid leukemia. Possible molecular mechanisms underlying the pathogenesis of the disease are discussed. PMID- 11408070 TI - Acute myeloid leukemia with concomitant trisomies 4 and 10: a distinctive form of myeloid leukemia? AB - The occurrence of trisomy 4 or trisomy 10 as the sole chromosomal abnormality in acute myeloid leukemia (AML) is very rare, the reported frequency being less than 1%. We describe two cases of AML-M2 with concomitant trisomy 4 and trisomy 10, a hitherto undescribed phenomenon. They showed two unusual features, including immunoreactivity for CD56 and a short-lived but rapidly progressive myelodysplastic phase preceding the appearance of frank leukemia. These findings raise the possibility that AML with concommitant trisomy 4 and trisomy 10 may constitute a distinctive subtype of AML. PMID- 11408071 TI - Trisomy 21 as the sole acquired karyotypic abnormality in an adult patient with CD7-positive acute myeloid leukemia. AB - We report a unique case of acute myeloid leukemia (AML) in which trisomy 21 was the sole acquired karyotypic abnormality. The blasts were positive for myeloperoxidase, and phenotypic analysis of peripheral blood cells by flow cytometry demonstrated positivity for CD7, CD13, CD33, and CD34. chromosomal analysis of peripheral blood and bone marrow cells showed trisomy 21; however, that of the buccal mucosal membrane revealed a normal karyotype. A diagnosis of CD7+AML (M2) with trisomy 21 was diagnosed and the patient achieved complete remission following treatment with Japan Adult Leukemia Study Group-AML97 protocol. This is the first reported case of CD7+AML with trisomy 21 as the sole cytogenetic abnormality. PMID- 11408072 TI - DNA copy number changes in familial malignant mesothelioma. AB - Malignant mesothelioma (MM) is predominantly a sporadic malignancy linked to exposure to asbestos. Clustering of MM in families suggests genetic susceptibility as a contributing factor. We performed comparative genomic hybridization (CGH) analysis on tumor samples from members of a family with MM of the pleura and a history of parental cancer. Our specific aim was to find a recurrent copy number loss indicating the chromosomal area to which a gene underlying the development of MM could be assigned according to the Knudson two hit hypothesis. We found losses at 1p, 6q, 9p, 13q, and 14q. The copy number changes were very similar to those reported in sporadic cases. Our findings and results from sporadic cases highlight the importance of cloning the genes in the loss sites at 1p, 6q, 14q, and 22q. PMID- 11408073 TI - Translocation (16;17)(q22;p13) is a recurrent anomaly of aneurysmal bone cysts. AB - Recently, Panoutsakopoulos et al. (1999) reported 2 cases of aneurysmal bone cysts with a recurrent (16;17)(q22;p13) translocation. We present here two additional cases harboring the same translocation as well as additional chromosomal changes. PMID- 11408074 TI - Gain of an isochromosome 5p: a new recurrent chromosome abnormality in acute monoblastic leukemia. AB - In acute myeloid leukemia (AML) close associations are known between cytomorphology and cytogenetics such as in AML M3/M3v showing a t(15;17) and in AML M4eo associated with inv(16)/t(16;16). In AML M5 a heterogenous cytogenetic pattern is observed. We describe the gain of an isochromosome of the short arm of chromosome 5 together with the gain of chromosome 8 as the sole abnormalities in two cases of acute monoblastic leukemia. In a third case of acute monoblastic leukemia we also observed the gain of an isochromosome 5p together with trisomy 8. This patient showed in addition an unbalanced translocation between the long arm of chromosome 1 and the short arm of chromosome 14 leading to a trisomy 1q. So far only two cases of AML with i(5)(p10) have been published. In no other hematological malignancy has an isochromosome 5p been reported up to now. As an isochromosome 5p can be misinterpreted as a deletion 5q, which occurs frequently in AML, fluorescence in situ hybridization with loci specific probes is a helpful method to detect this rare abnormality. PMID- 11408075 TI - Hyperdiploidy in a case of favorable histology Wilms tumor. AB - Hyperdiploidy is useful in defining histologic variants of Wilms tumor and prognosis in other childhood cancers. We describe a case of hyperdiploid favorable histology Wilms tumor (50,XY,+6,+7,+8,+10,+12, 21[2]/51,idem,+9[6]/46,XY[12]) in a 3-year-old boy, and review the literature for other hyperdiploid childhood renal lesions. PMID- 11408076 TI - Cellular approaches for diagnostic effects assessment in ecotoxicology: introductory remarks to an EU-funded project. PMID- 11408077 TI - Metabolic fate of 2,4-dichloroaniline, prochloraz and nonylphenol diethoxylate in rainbow trout: a comparative in vivo/in vitro approach. AB - The metabolism and distribution of 2,4-dichloroaniline (2,4-DCA), prochloraz and 4-n-nonylphenol diethoxylate (NP2EO) were investigated in vivo and in vitro in rainbow trout (Oncorhynchus mykiss). Each compound was administered p.o. (10 mg/kg wet weight) and urine was collected during 48 h (2,4-DCA, prochloraz) or 72 h (NP2EO). Fish were sacrificed, the gall bladder was excised and radioactivity was measured in tissues, viscera and carcasses. Metabolic profiles were performed by radio-HPLC and when possible metabolites were identified by LC/MS. For comparison, the biotransformation of these xenobiotics was also investigated in freshly isolated hepatocytes. The metabolic pathways of 2,4-DCA have been identified leading to the glucuronide conjugate (in vivo) and to the glucuronide conjugate and the hydroxylamine metabolite (in vitro). This difference highlights the usefulness of the hepatocyte system in metabolic studies, since the formation of the hydroxylamine reactive metabolite cannot be demonstrated in vivo. For prochloraz, we observed that residue levels are significantly higher in males than in females for gill, fat, brain and carcasses, however, the reasons for this difference remain unclear. Although, the presence of glucuronide conjugates was detected in vivo and in vitro, the chemical structure of isolated metabolites has to be determined. However, the comparison of the in vivo versus in vitro metabolic profiles indicates that several peaks, probably corresponding to intermediate metabolites, were present only in hepatocyte incubations. Biotransformation of NP2EO occurred in vivo and in vitro in rainbow trout, but did not result in the formation of 4-n-NP. The major metabolite present in bile corresponded to the NP2EO-glucuronide but this metabolite was not found in vitro. It is concluded that hepatocytes may produce a different metabolic pattern than in the whole fish, but may also give evidence of a metabolic pathway difficult to apprehend in vivo. PMID- 11408078 TI - Effects of nonylphenol on estrogen receptor conformation, transcriptional activity and sexual reversion in rainbow trout (Oncorhynchus mykiss). AB - Estrogenic potency of 4-n-nonylphenol diethoxylate, 4-n-nonylphenol (NP) and metabolites were tested using two bioassays: rainbow trout hepatocyte culture and recombinant yeast stably expressing rainbow trout estrogen receptor (rtER) and containing estrogen-dependent reporter genes. Since NP was the only compound active in both systems, its interaction with rtER was studied in more detail. Qualitative and quantitative differences were observed in the presence of 17beta estradiol (E2) or NP when estrogen-dependent promoters containing one to three estrogen-responsive elements were used in yeast. Moreover, limited proteolysis of rtER after E2 or NP binding presented different patterns after SDS-PAGE analysis suggesting that NP induces a differential conformation of rtER compare to E2. This finding may have important implications with respect to the biological activity of NP. Thus, the effects of NP on the activation of an E2-dependent gene and on sexual differentiation were assessed on all-male trout embryos exposed to NP for 1 h per day for 10 days. Although in situ hybridization demonstrated that E2, and to a lesser extend NP, were able to increase rtER mRNA level in the liver of embryos, no indication of total or partial sexual reversion was observed (even in E2 treated fishes) when the gonads were examined 8 months after hatching. PMID- 11408079 TI - In vivo and in vitro effects of prochloraz and nonylphenol ethoxylates on trout spermatogenesis. AB - We investigated the effects of in vivo exposure to non-lethal concentrations of two chemicals commonly discharged into the aquatic environment, prochloraz and nonylphenol diethoxylate (NP2EO - Igepal(R) 210), on the development of spermatogenesis in trout. The in vitro effects on basal and insulin-like growth factor-1 (IGF-I) stimulated DNA synthesis by early germ cells were also studied. In vivo, rainbow trout were exposed for 2 or 3 weeks to waterborne prochloraz (21 and 175 nmol/l) and/or NP2EO (68-970 nmol/l) renewed continuously, or periodically. Only the highest concentrations of NP2EO (225-970 nmol/l) induced a significant increase in blood plasma vitellogenin in juvenile or maturing male trout. When prepubertal fish were exposed for 15 days to prochloraz, the spermatogenetic process was significantly inhibited as shown by the stage of gonadal development reached 3 weeks after exposure. This effect was, to a great extent, reversible within 9 weeks post-exposure. When fish in the initial stage of spermatogenesis were exposed for 21-27 days to 580 nmol/l NP2EO, a 20-40% reduction of the gonadosomatic index was observed 4.5 weeks post-exposure, and the spermatogenetic process was partly inhibited. In vitro, testicular cells obtained at different stages of spermatogenesis were cultured for 4.5 days in the presence or not of the tested molecules and with IGF-I or not. 3H-thymidine (3H Tdr) incorporation was measured according to Loir (Mol. Reprod. Dev. 53 (1999) 424) and 125I-IGF-I specific binding was determined according to Le Gac et al. (Mol. Reprod. Dev. 44 (1996) 35). Irrespective of the spermatogenetic stage, basal 3H-Tdr incorporation was decreased by prochloraz concentrations > or =10 micromol/l. The presence of IGF-I (10-100 ng/ml) stimulated 3H-Tdr incorporation; this response to IGF-I began to decrease at 25-50 micromol/l prochloraz. In parallel, a dose-dependent increase of IGF-I specific binding was induced by prochloraz 1-100 micromol/l. Similarly, basal and IGF-I-stimulated 3H-Tdr incorporation was decreased by nonylphenol polyethoxylate (NpnEO; starting at 10 micromol/l), NP2EO and NP (30 micromol/l); a dose-dependent increase of IGF-I specific binding was also induced by NP and NPnEO. While 1-100 nmol/l 17beta estradiol had no effect in our in vitro system, Triton(R) X-100 acted as NPnEO on 3H-Tdr incorporation. Beside their known endocrine disrupting effects on sex steroid production or action, these lipophilic molecules could act on germ cells by disrupting cell membrane receptivity to peptide hormones like growth factors. PMID- 11408080 TI - In vitro effect of various xenobiotics on trout gill cell volume regulation after hypotonic shock. AB - Their functions and localisation can expose gill cells to volume changes. To maintain their vital functions, these gill cells must regulate their own volume after cellular swelling or shrinkage. Recently, we showed that rainbow trout pavement gill cells in primary culture have the capacity to regulate their own volume after cellular swelling induced by hypotonic shock. This so-called regulatory volume decrease (RVD) is associated with intracellular calcium increase, which occurs as a transient peak followed by a plateau when maintained a hypotonic condition. Return to an isotonic medium restores baseline [Ca2+]i level. In this study, the effect of different xenobiotics on cellular swelling induced RVD and its calcium signal was investigated in trout pavement gill cells in primary culture. These cells were exposed to different pollutants after confluent epithelium was obtained. After 36 h in xenobiotics exposure in vitro, cellular volume and intracellular calcium concentration were measured. Nonylphenol poly- and di-ethoxylate were lethal at concentrations of 10 and 100 microM, respectively. With 10 microM of the diethoxylate form, cells did not die but, unlike non-treated cells, burst during hypotonic shock (2/3rd strength Ringer solution). With 1 microM nonylphenol polyethoxylate (NPnEO), RVD and [Ca2+]i were reduced. Copper (10 and 100 microM) had no significant effect on gill cell volume regulation. However, the heavy metal modified calcium response to hypotonic shock by inhibiting return to baseline level under isotonic conditions. 10 microM prochloraz and 2,4-dichloroaniline had no effect on cell morphology, volume and [Ca2+]i concentration. With 100 microM, however, prochloraz was lethal and dichloroaniline increased baseline [Ca2+]i. These results indicate that the effects observed on gill cells are consistent with the known toxic properties of the molecules tested, thus confirming the validity of primary culture to investigate the toxic effects of xenobiotics on fish gill epithelium. PMID- 11408081 TI - Prochloraz and nonylphenol diethoxylate inhibit an mdr1-like activity in vitro, but do not alter hepatic levels of P-glycoprotein in trout exposed in vivo. AB - P-glycoproteins (P-gps) encoded by multidrug resistance 1 (mdr1) genes are ATP dependent transporters located in the cytoplasmic membrane which mediate the efflux of a broad spectrum of hydrophobic compounds from the cell. The tissue distribution of P-gps suggests their role in the organismal defense against xenobiotics by effecting xenobiotic excretion and reducing xenobiotic uptake. In the present work, the interaction of P-gp(s) in the liver and in primary cultured hepatocytes of rainbow trout with two model pollutants was studied - the imidazole fungicide prochloraz and the alkylphenolic surfactant nonylphenol diethoxylate (NP2EO). Using a monoclonal antibody (mAB C219) directed against a conserved P-gp epitope, an immunoreactive protein of 160 kDa was detected in immunoblots of liver extracts from control trout. In sections of control trout livers, immunohistochemistry with the mAB C219 resulted in specific staining of bile canaliculi. In juvenile trout exposed for 7 days to sublethal concentrations of prochloraz (0.027 microM; 0.27 microM) or NP2EO (0.32 microM; 1.30 microM), no changes in levels of hepatic P-gp(s) were found in immunoblot and immunochemical investigations. The efflux of the fluorescent mdr 1 substrate rhodamine 123 (Rh123) from cultured isolated trout hepatocytes was partly inhibited by verapamil and vinblastine, compounds known to interfere with mdr 1-dependent transport. This demonstrates the presence of a mdr1-like mechanism in trout liver which is probably involved in the biliary excretion of hydrophobic xenobiotics. Non-cytotoxic concentrations of prochloraz and NP2EO were tested for effects on the efflux of Rh123 from trout hepatocytes. Prochloraz was a potent inhibitor of the mdr1-like mechanism, being effective at 0.3 microM and above. NP2EO inhibited Rh123 efflux only at the highest concentration tested (31.6 microM). The accumulation and elimination of 14C-prochloraz by cultured trout hepatocytes was not affected by mdr 1-type substrates (Rh123, vinblastine) and a mdr 1 inhibitor (verapamil). This shows that prochloraz is, despite its inhibitory potency, not a substrate of the mdr1-like mechanism in trout liver. The inhibition by prochloraz and NP2EO of the md r1-like mechanism in trout hepatocytes suggests that water pollutants can interfere with P-gp-function in fish and thus may impair the organismal defense against xenobiotics. PMID- 11408082 TI - Effects of prochloraz and nonylphenol diethoxylate on hepatic biotransformation enzymes in trout: a comparative in vitro/in vivo-assessment using cultured hepatocytes. AB - The suitability of cultured rainbow trout hepatocytes as a model system for the assessment of xenobiotic effects on hepatic biotransformation enzymes in fish was examined. Two model water pollutants, the imidazole fungicide prochloraz and the alkylphenolic compound nonylphenol diethoxylate (NP2EO), were investigated in a comparative in vitro/in vivo approach. Biotransformation enzymes were measured in cultured rainbow trout hepatocytes following exposure to xenobiotics in vitro, or in the liver of juvenile rainbow trout (Oncorhynchus mykiss) exposed in vivo. The patterns of biochemical responses to the model pollutants were generally similar between in vitro and in vivo investigations. Levels of cytochrome P4501A (CYP1A) protein and the catalytic activity of the CYP1A-dependent enzyme 7 ethoxyresorufin-O-deethylase (EROD) were induced in vitro after 24 h of exposure to 1.0 microM prochloraz. In vitro, higher prochloraz concentrations induced only the levels of CYP1A above control levels, but not EROD activity. In vivo exposure of juvenile trout to 0.27 microM prochloraz resulted in an induction of CYP1A and EROD after 7 and 14 days, while 0.027 microM prochloraz had no effects. In vitro, the 6beta- and 16beta-hydroxylation of testosterone was significantly decreased by 1.0-3.0 microM prochloraz, while in vivo these variables were significantly inhibited after exposure to 0.27 microM prochloraz for 7 and 14 days. NP2EO did not affect EROD activity in vitro. In vivo, EROD activity and CYP1A remained unchanged following 7 days of exposure to 0.32 or 1.30 microM NP2EO. NP2EO (15-50 microM) inhibited the 16beta-hydroxylation and glucuronidation of testosterone in vitro. In vivo, 7 days of exposure to 0.32 or 1.30 microM NP2EO resulted in a significant inhibition of the 6beta- and 16beta-hydroxylation of testosterone. The good qualitative correspondence between in vitro and in vivo results indicates that studies using trout hepatocytes allow the identification of biochemical targets of xenobiotic effects in fish liver. However, more research is needed before quantitative predictions, e.g. of effective concentrations, can be made from in vitro investigations. PMID- 11408083 TI - Sensitivity of muscle satellite cells to pollutants: an in vitro and in vivo comparative approach. AB - Muscle satellite cells from rainbow trout were exposed in vitro to increasing concentrations of different xenobiotics: copper, dichloroaniline, prochloraz, nonyl-phenol polyethexylate. Mortality and proliferation rate were measured by Hoechst binding and BrdU incorporation. Dose dependent effect of copper on survival and proliferation was observed with an EC50 at 100 microM. A dose dependent effect of nonyl-phenol diethoxylate was observed on survival with an EC50 at 100 microM. This was associated with a biphasic effect on proliferation rate observed both for nonyl-phenol di and poly-ethoxylate: a stimulation of proliferation at low concentration and an inhibition proliferation at large concentration. These effects were related to the inhibition of cells adhesion through the detergent capacity of nonyl-phenol polyethoxylate. The effects of prochloraze and dichloroaniline on cells mortality (EC 50 > 500 microM) and proliferation rate (LOEC: 100 microM) were limited. Whole fish growth, muscle fibre size distribution and satellite cells survival and proliferation were measured on rainbow trout (60-80 g BW) exposed to two concentrations of prochloraze (10 and 100 microg/ml) or nonyl phenol diethoxylate IGEPAL 210 (100 and 400 microg/ml) during 14 and 10 days exposure, respectively. Muscle fibre size distribution and satellite cells proliferation were affected by prochloraz exposure in vivo and this could be related to the alteration in fish feeding status. The exposure to IGEPAL 210 affected the number of satellite cells extracted and induce a biphasic effect on satellite cells proliferation similar to that observed in vivo. The combined exposure to IGEPAL 210 and prochloraze revealed additive effects of these two compounds. The in vivo and in vitro comparison demonstrated that in vitro satellite cells system could be used as a valuable tool to test the effects of pollutants. PMID- 11408084 TI - Protein synthesis costs could account for the tissue-specific effects of sub lethal copper on protein synthesis in rainbow trout (Oncorhynchus mykiss). AB - This study investigates protein synthesis, following exposure to sub-lethal Cu, in rainbow trout in vivo and in vitro. The investigation has two aims: to determine if perturbations in protein synthesis, compared with other physiological changes, are a biomarker of Cu pollution and to evaluate the most productive role of cellular models in ecotoxicology. Protein synthesis rates were measured by labelling with 3H-phenylalanine. In vivo this was applied by a single (i.p.) injection and in vitro by bathing the cells in 3H-phenylalanine labelled culture media. The effects in vivo were tissue specific. After 3 weeks' exposure to 0.7 microM Cu only skin protein synthesis was reduced. Gills and liver from the same fish were unaffected. This reduction in skin protein synthesis appears to be more sensitive than some other biomarkers reported in the literature. However, Cu concentrations greater by orders of magnitude were required to reproduce this reduction in protein synthesis in skin cell explants (200 and 400 microM). Hepatocyte protein synthesis was unaffected by 10, 20 and 40 microM Cu and a separate investigation has also shown that 25 and 75 microM Cu does not effect protein synthesis in cultured gill cells. Oxygen consumption rates were also measured in vitro by monitoring the decline in O2 partial pressure. The Cu concentrations given above resulted in a decline in O2 consumption rates in the respective cell types. By measuring protein synthesis and O2 consumption after treatment with a protein synthesis inhibitor (cycloheximide), the costs of protein synthesis were also determined. Synthesis costs in hepatocytes are close to the theoretical minimum and are only marginally affected by Cu. Gill cell synthesis costs are also minimal and are unaffected. In skin explants, the reduction in protein synthesis was accompanied by greatly increased synthesis costs. This in vitro result offers a hypothesis as to the tissue-specific effects in vivo; i.e. the energetic demand of protein synthesis may determine tissue sensitivity or susceptibility. Cell or tissue types with high protein synthesis rates are able to avoid detrimental increases in the synthesis cost when exposed to Cu. In tissues with a low protein synthesis rate any further reduction is more likely to incur a potentially damaging increase in protein synthesis costs. Thus, whilst in vitro models may have little practical use in environmental monitoring, they may be best used as a mechanistic tool in understanding susceptibility or tolerance to sub-lethal Cu. PMID- 11408085 TI - The further development of rainbow trout primary epithelial cell cultures as a diagnostic tool in ecotoxicology risk assessment. AB - The use of short-term cytotoxicity assays for the initial screening of chemicals not only aids in establishing priorities for the selection of chemicals that should be tested in vivo, but also decreases the time in which potential toxicants can be valued. Rainbow trout primary skin epithelial cell cultures are one such assay. Rainbow trout primary skin cell cultures contain two cell types, keratinocytes and goblet mucus cells. Two aquatic pollutants, copper and prochloraz were screened using this cell system. The influence of media composition on the effects of the aquatic pollutants was also studied by testing the chemicals in both serum-containing and serum-free medium and the morphological changes that occurred within the cell cultures recorded. The concentration of copper that causes a reduction of 90% in the residual of day 3 growth of the primary cell culture system was found to be approximately 10 fold more than that of prochloraz. Prochloraz was found to cause a greater reduction in growth area when added to the primary cell culture system in serum-free medium than in serum-containing medium. Copper, in contrast, was found to exert reduced toxicity when added to the test cultures in serum-free medium compared with addition in serum-containing medium. Prochloraz was found to kill the epithelial cells by a process of necrosis. Copper, was found to kill the epithelial cells by both necrosis and apoptosis in a ratio of 2:1. It was also observed that as the dose of both chemicals increased, the number of goblet cells contained in the cell cultures decreased. A PAS stain was carried out to determine if the goblet cells were exocytosing their contents onto the cell culture surface. It was found that as chemical exposure increased the number of cells expressing positivity for mucus also increased. The results of this study add further evidence to support that primary cell cultures are a very appropriate model for toxicity risk assessment. PMID- 11408086 TI - Ricin toxicity to microglial and monocytic cells. AB - Microglial cells, like macrophages, are very sensitive to ricin, a galactose specific toxic lectin belonging to the family of ribosome-inactivating proteins. This toxin can be taken up by most cells through the binding of its B chain to galactose-containing molecules on the cell membrane. In macrophagic cell types it can be internalised also by mannose receptors which are present on the surface of these cells. Endocytosis of the toxin by either pathway was evaluated by ricin toxicity to primary cultures of rat microglial cells and to a microglial N11 cell line in the presence or absence of lactose and mannan, which compete for the endocytosis via the ricin lectin chain or cellular mannose receptors, respectively. Results were compared with those obtained in cultures of mouse macrophages, human monocytes, and a monocytic JM cell line. All cultures were protected from ricin toxicity more by lactose than by mannan, indicating that ricin endocytosis via its lectin B chain is prevalent over that mediated by cellular mannose receptors. However, a partial protection by mannan was observed in all cases but not-stimulated N11 cells, either in the form of direct protection or of significant additional protection over that afforded by lactose. Mannose receptor expression by N11 cells was negative before, and positive after, treatment with endotoxin, as assessed by the specific binding of 125I-mannose bovine serum albumin. Moreover, a partial protection from ricin toxicity by mannan was induced in the N11 microglial line after stimulation, consistently with an inducible expression of the mannose receptor by activated cells switched towards a microglial phenotype. PMID- 11408087 TI - Melatonin activates PKC-alpha but not PKC-epsilon in N1E-115 cells. AB - Previous reports have revealed that calmodulin antagonism by melatonin is followed by microtubule enlargements and neurite outgrowths in neuroblastoma N1E 115 cells. In addition, activation of protein kinase C (PKC) by this neurohormone is also followed by increased vimentin phosphorylation, and reorganization of vimentin intermediate filaments (IFs) in N1E-115 cells. In this work, we further characterize the activation of PKC by melatonin in neuroblastoma N1E-115 cells. We studied the Ca(2+)-dependent effects of melatonin on PKC activity and distribution of PKC-alpha in isolated N1E-115 cell IFs. Also, the effects of melatonin on PKC-alpha translocation in comparison to PKC-epsilon, were studied in intact N1E-115 cells. The results showed that both melatonin and the PKC agonist phorbol-12-myristate-13-acetate increased PKC activity in isolated IFs. The effects of the hormone were Ca(2+)-dependent, while those caused by the phorbol ester were produced with or without Ca(2+). Also, in isolated in situ IFs, the hormone changed the distribution of PKC-alpha. In intact N1E-115 cells, melatonin elicited PKC-alpha translocation and no changes were detected in PKC epsilon. Phorbol-12-myristate-13-acetate modified the subcellular distribution of both PKC isoforms. The results showed that melatonin selectively activates the Ca(2+)-dependent alpha isoform of PKC and suggest that PKC-alpha activation by melatonin underlies IF rearrangements and participates in neurite formation in N1E-115 cells. PMID- 11408088 TI - Further evidence for a role of NMDA receptors in the locus coeruleus in the expression of withdrawal syndrome from opioids. AB - To examine a role of N-methyl-D-aspartate (NMDA) receptors in the locus coeruleus (LC) in the expression of the withdrawal signs from opioids, rats were continuously infused with morphine (a mu-opioid agonist, 26 nmol/microl per h) or butorphanol (a mu/delta/kappa-mixed opioid agonist, 26 nmol/microl per h) intracerebroventricularly (i.c.v.) through osmotic minipumps for 3 days. An LC injection of NMDA (0.1 and 1 nmol/5 microl) induced withdrawal signs in opioid dependent animals. However, it did not precipitate any abnormal behaviors in saline-treated control rats. The expression of the withdrawal signs precipitated by NMDA (1 nmol/5 microl), glutamate (10 nmol/5 microl), or naloxone (an opioid antagonist, 24 nmol/5 microl) was completely blocked by pretreatment with a NMDA antagonist, MK-801 (5-methyl-10,11-dihydro-5H-dibenzo[a,d]cycloheptan-5,10 imine), 0.1 mg/kg, i.p. In animals that had been infused with opioids in the same manner, naloxone (48 nmol/5 microl, i.c.v.) precipitated withdrawal signs and increased extracellular glutamate levels in the LC of opioid-dependent rats measured by in vivo microdialysis method. Pretreatment with MK-801, however, did not affect the increases of glutamate levels in the LC. These results further demonstrate that the expression of opioid withdrawal induced by an expeditious release of glutamate in the LC region of opioid-dependent animals might be mainly mediated by the postsynaptic NMDA receptors. PMID- 11408089 TI - Little change in cerebrospinal fluid amino acids in subtypes of multiple sclerosis compared with acute polyradiculoneuropathy. AB - Levels of free amino acids were determined in randomised, blinded samples of cerebrospinal fluid (CSF) from patients with relapsing-remitting or chronic progressive multiple sclerosis (MS), all in the active phase of disease. The levels were compared with amino acid amounts in patients with an acute polyradiculoneuropathy (Guillain-Barre syndrome (GBS)) and a control population of patients with no known neurological disease or deficit. The data did not indicate any significant changes in amino acid levels between MS subgroups. The only significant differences between MS patients and controls were a modest reduction in glutamate and a slight increase in taurine, but the changes were so small that the biological relevance is dubious. These results contrasted with the marked increases for many amino acids in CSF from patients with acute polyradiculoneuropathy compared with controls. The amino acid profile in cerebrospinal fluid (CSF) does not appear to provide evidence of differential pathology in multiple sclerosis (MS). The increase in hydrophobic amino acids and lysine in CSF from patients with acute polyradiculoneuropathy is consistent with transudation over the blood-CSF barrier following an infection. The increases in glutamine and alanine may reflect increased nitrogen removal from brain. PMID- 11408090 TI - Dopamine D1-stimulated adenylyl cyclase activity in cerebral cortex of autopsied human brain. AB - Although the cerebral cortical dopamine D(1) receptor is considered to play a role in normal and abnormal brain function, little information is available on its characteristics in human brain. We compared dopamine-stimulated adenylyl cyclase (AC) activity in homogenates of cerebral cortex (frontal, temporal, parietal, occipital and cingulate cortex) of autopsied brain of neurologically normal subjects to that in striatum. Cerebral cortical AC activity was modestly and dose-dependently stimulated by dopamine (maximal 20-30%) with low microM EC50s and such stimulation was inhibited by the selective dopamine D1 receptor antagonist SCH23390. The magnitude of the maximal stimulation by dopamine was similar in autopsied and biopsied cerebral cortex. The extent of maximal stimulation was similar to that in dopamine-rich striatum (caudate, putamen and nucleus accumbens), despite much lower density of dopamine D1 receptors in cerebral cortex vs. striatum. The EC50 for dopamine stimulation in cerebral cortex (approximately 1 microM) was lower than that for caudate and putamen (approximately 3 microM). No detectable dopamine stimulation was observed in cerebellar cortex, thalamus or hippocampus. Dopamine stimulation in both cerebral cortex and striatum was independent of calcium activation. We conclude that dopamine stimulated AC can be measured in cerebral cortex of human brain allowing for the possibility that this process can be examined in human brain disorders in which dopaminergic abnormalities are suspected. PMID- 11408091 TI - Lipid peroxidation-mediated oxidative stress and dopamine neuronal apoptosis in the substantia nigra during development. AB - The cellular pathways underlying naturally occurring neuronal apoptosis in the rat substantia nigra (SN) during the perinatal period remain largely unknown. Determining the mediators of this process in development may shed light on causes of premature neuronal death in adult neurodegenerative disorders, including the loss of dopamine neurons in Parkinson's disease. In the present study, we investigated whether lipid peroxidation-mediated oxidative stress mediates developmental death of nigral neurons by (1) establishing the profile of lipid peroxidation and other oxidative stress markers throughout the postnatal period both in the SN and striatum, and (2) examining whether the inhibitor of lipid peroxidation, alpha-tocopherol, protects these neurons from death. In addition to monitoring, the level of lipid peroxidation throughout development, we also measured the activities of three antioxidant enzymes, namely superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx). We have shown that lipid peroxidation and SOD activity progressively increased from postnatal day (PND) 3 to PND 42 in both SN and striatum. During this period, GPx activity remained stable, while catalase activity transiently increased at PND 8 only in the SN. Furthermore, alpha-tocopherol treatment from embryonic day 18 to PND 2 did not reduce the number of apoptotic neurons at PND 3. These results do not support the hypothesis that lipid peroxidation-mediated oxidative stress is the major mediator of nigral dopamine neuronal apoptosis during the perinatal period. PMID- 11408092 TI - Regional distribution of ornithine decarboxylase activity and polyamine levels in experimental cat brain tumors. AB - Biosynthesis of the polyamines putrescine, spermidine, and spermine, and activation of the first key enzyme ornithine decarboxylase (ODC) are closely associated with cellular proliferation. In the present study, the distribution of ODC activity and polyamine levels was investigated for the first time regionally in experimental brain tumors of the cat. Brain tumors were produced by stereotactic xenotransplantation of rat glioma cells. Twenty days after implantation, the brains were frozen in situ, cut into slices, and cryostat sections and tissue samples were taken to determine ODC activity and polyamine levels biochemically. The quantified data were color-coded to present the regional distribution of ODC activity and polyamine levels in the respective section. ODC activity significantly increased in some areas within the tumor, whereas peritumoral tissue showed no difference to the non-tumoral, contralateral hemisphere. This increase turned out in parallel to a high number of mitoses in the same tumor parts (r=0.861). Putrescine levels increased both, in the whole tumor and in the peritumoral edema. Regional differences in putrescine content did not correlate with solid and proliferative parts of the tumor. Spermidine and spermine levels were only slightly increased in some parts of the tumor. Thus, these experiments show the close correlation of a high mitotic rate and activation of ODC within experimental gliomas and underline the relevance of ODC as a biochemical marker of proliferation in brain tumors. PMID- 11408093 TI - Glutathione elevation and its protective role in acrolein-induced protein damage in synaptosomal membranes: relevance to brain lipid peroxidation in neurodegenerative disease. AB - Oxidative stress may be a hallmark of several neurodegenerative disorders, including Alzheimer's disease (AD) Huntington's, and Parkinson's diseases as well as amyotrophic lateral sclerosis. Acrolein is a highly reactive product of lipid peroxidation that is elevated in the brains of persons with AD. This alkenal potentially can react with proteins by Michael addition to alter their structure and function. In the present study, we used electron paramagnetic resonance in conjunction with a protein-specific spin label to monitor synaptosomal membrane protein conformational alterations induced by acrolein. A dose-dependent increased conformational alteration was observed. Consistent with this finding, protein carbonyl levels from protein-bound acrolein were significantly elevated. However, pretreatment of synaptosomes with glutathione ethyl ester (GEE) significantly ameliorated both the conformational alterations and protein carbonyls induced by acrolein. Based on this success, we tested the hypothesis that elevated levels of endogenous glutathione (GSH) would offer protection against acrolein-induced oxidative stress. In-vivo elevation of GSH (215% over control, P<0.04) was produced by i.p. injection of N-acetylcysteine (NAC), a known precursor of GSH. Synaptosomes were treated with vehicle or 2 nM acrolein, the level of this alkenal found in AD brain. In contrast to synaptosomes from control animals, which had significantly increased protein carbonyl levels following addition of 2 nM acrolein, synaptosomes that were isolated from NAC treated rodents and treated with 2 nM acrolein showed no increased carbonyl levels compared to untreated controls. These results demonstrate protection by increased in-vivo GSH levels against acrolein-induced oxidative stress at levels found in AD brain and are consistent with the notion that methods to increase endogenous GSH levels in neurodegenerative diseases associated with oxidative stress may be promising. PMID- 11408094 TI - Increased ambient glutamate concentration alters the expression of NMDA receptor subunits in cerebellar granule neurons. AB - Effects of prolonged (48 h) inhibition of glutamate reuptake on the relative abundance of mRNAs coding for N-methyl-D-aspartate (NMDA) receptor subunits, and the expression of corresponding proteins were investigated in primary cultures of rat cerebellar granule neurons. In cells exposed to the glutamate transport blocker, L-trans-pyrrolidine-2,4-dicarboxylate (PDC), the expression of the C1 exon-positive NR1 mRNA variant was reduced by about 40% whereas, the expression of C1-negative mRNA was increased leading to significant reduction of the +C1/-C1 ratio. The expression of the N1-negative NR1 variants was slightly reduced following exposure to PDC, indicating that increased medium levels of glutamate changed the relative abundance of NR1 splice-variant expression but did not reduce the overall NR1 transcription. Expression of NR2A and NR2B mRNAs was 40 50% lower in PDC-treated cells as compared to control. Immunoblot experiments revealed that PDC exposure reduced the expression of NR1 and all NR2 proteins with NR2A and NR2B proteins being reduced to a greater extent than NR1. The overall decrease in NMDA receptor subunit protein expression was considerably more pronounced than the reduction of their corresponding mRNAs, suggesting involvement of a post-transcriptional regulation. Our data support the hypothesis that functional activity and number of NMDA receptors are regulated by strength of the glutamatergic input. Thus, reduced glutamate uptake resulting in increased concentration of ambient glutamate initiate a series of adaptive responses manifested as a gradual down-regulation of the functional activity and expression of NMDA receptors. PMID- 11408095 TI - The development of dopamine overflow from foetal nigral grafts in the intact rat striatum and their influence on contralateral striatal dopamine overflow. AB - In this study, cell suspensions of foetal rat ventral mesencephalic dopaminergic tissue were grafted to the intact (non-lesioned) striatum of adult rats. Differential pulse voltammetry at carbon-fibre micro electrodes (12 microm diameter) was employed to first, monitor the development of dopamine overflow over a 20 week period within the grafts and secondly, their influence on contralateral striatal dopamine overflow. At 8 and 20 weeks, animals were pre treated with pargyline and both striata were monitored for dopamine overflow for 90 min following d-amphetamine administration. Amphetamine led to a significant increase in dopamine overflow in both the grafted striatum and the contralateral striatum. The time course of dopamine overflow in both the grafted striatum and the striatum contralateral to the graft was similar in all groups of animals. Although the actual concentration of dopamine measured in 20 week old grafts was more (approximately 21%) than that measured in 8 week old grafts, there was no significant difference between the two time points. The concentration of dopamine measured in the striatum contralateral to 8 week old grafts was significantly lower (approximately 43%) than that measured in the striatum of a normal control rats. There was no significant difference between the concentration of dopamine measured in the striatum contralateral to 20 week old grafts and normal control rats. In conclusion, dopamine overflow from a ventral mesencephalic graft does not change significantly between 8 and 20 weeks following grafting. However, the grafted tissue causes a decrease of d-amphetamine-induced dopamine overflow in the contralateral side 8 weeks following grafting, which is restored 12 weeks later. PMID- 11408096 TI - Pathologic and laboratory correlation in microcephaly associated with prenatal cocaine exposure. AB - The Authors report a case where cocaine abuse during pregnancy assessed by drug analysis at various site was associated with foetal microcephaly. Foetal pathologic findings revealed anomalies in neuronal migration and in the vascular architecture in the brain. Such anomalies might be the result of prolonged exposure to cocaine in utero, aggravated by the high concentration of cocaine metabolites in the amniotic fluid over a prolonged period. PMID- 11408097 TI - Neurological outcome in school-age children after in utero exposure to coumarins. AB - The effect of prenatal exposure to coumarins (acenocoumarol, phenprocoumon) on neurological outcome was assessed in a cohort of 306 children aged 7-15 years. Findings were compared with those in a non-exposed cohort of 267 children, matched for sex, age, and demographic region. We used a neurological examination technique which pays special attention to minor neurological dysfunction (MND). None of the children was found to be neurologically abnormal. However, exposure to coumarins during gestation increases the risk for MND in children of school age, odds ratio (OR) 1.9 (CI(95) 1.1-3.4), predominantly after exposure in the second or third trimester, odds ratio 2.1 (CI(95) 1.2-3.8). We found a dose response relationship with an odds ratio of 1.2 (CI(95) 1.0-1.5) per mg coumarin derivative prescribed per day. The results suggest that coumarins have an influence on the development of the brain which can lead to mild neurological dysfunctions in children of school age. PMID- 11408098 TI - Symphysis-fundal measurement as a predictor of low birthweight. AB - OBJECTIVE: To prospectively test the value of a symphysis-fundal measurement in labor of less than or equal to 29 cm as a predictor of birth mass below 2000 g. METHOD: Women admitted to Kalafong Hospital in labor with a singleton pregnancy were included in the study. Symphysis-fundal measurement according to the method of Westin was performed on two separate occasions and recorded together with other pertinent details. A receiver-operator curve was constructed to test various cut-off measurements. RESULTS: A total of 1216 women were included in the study. One hundred twenty-one fetuses had a birth mass less than 2000 g (10%). A measurement of less than or equal to 29 cm had a sensitivity of 69% and a specificity of 98% with a positive predictive value of 81% and a negative predictive value of 97%. On the receiver-operator curve a cutoff of 30 cm showed a better sensitivity with little loss of specificity. CONCLUSION: A symphysis fundal measurement of less than or equal to 29 cm is a good predictor of birth mass less than 2000 g and can be used as an indication for referral to centres with neonatal facilities. PMID- 11408099 TI - Selenium levels in kidney, liver and heart of newborns and infants. AB - BACKGROUND: Among the tissues of humans the highest level of selenium (Se) is found in the kidney and the lowest in the muscle. The Se level in tissues is age dependent. AIM: To measure the Se level in kidney, liver and heart of newborns and infants who were born in different periods of pregnancy and died of various diseases. SUBJECTS: Tissues obtained from 49 infants deceased at 1 day to 2.5 months of age. Forty-five of them were premature infants born between 23 and 36 weeks of pregnancy, four were born at term. RESULTS: Se levels in kidney and heart (but not liver) increased gradually with the duration of pregnancy. Positive and significant correlations were found between the weeks of pregnancy and Se levels in kidney (r=0.433, P=0.023) and heart (r=0.313, P=0.030). In the total group, the mean Se levels in the kidney (185+/-64.7 ng/g wet weight) and liver (177+/-59.8 ng/g) were two times higher than those in the heart (90.5+/ 33.7 ng/g). In this study, the Se levels in kidney and liver were from 2.5 to 1.2 times lower than the values found in our previous study for healthy adult victims. In the heart, the Se levels were similar in infants and adults. No significant differences were found in the mean Se levels in the various tissues in infants who died due to respiratory distress syndrome, congenital heart disease, other diseases and the group as a whole. The low levels of Se in the tissues studied by us, as compared with data from other countries, are probably due to lower Se intake by pregnant women in Poland. CONCLUSIONS: This study demonstrates that Se level in kidney and heart, but not in liver, increases with the progress of pregnancy. The Se level in kidney and liver is two times higher compared with heart and significantly higher than in adult subjects. PMID- 11408100 TI - Effect of antenatal corticosteroids on postmortem brain weight of preterm babies. AB - OBJECTIVES: To investigate the effects of single and repeated courses of antenatal corticosteroids on brain growth in very preterm babies. DESIGN: Retrospective study of 110 very preterm babies delivered at a single University Teaching Hospital between 1992 and 1999 who had a full necropsy including detailed examination of the brain. RESULTS: Mean brain weight did not differ significantly between babies who received corticosteroids and those who did not 160 vs. 157 g, (p=0.82), nor was there a difference between mean brain weight of stillborn or liveborn babies in relation to steroid use 164 vs. 159 g, (p=0.84) and 156 vs. 152g (p=0.81). There was no apparent dose-response relationship between the total number of doses of corticosteroids or timing since the first dose of corticosteroids and brain weight, p=0.95 and p=0.87. CONCLUSIONS: Single and multiple courses of antenatal corticosteroids had no significant effect on brain growth in babies delivered preterm who died but long-term follow-up studies are required to evaluate the functional neurological outcome of surviving children. PMID- 11408101 TI - Diurnal variation of eye movement and heart rate variability in the human fetus at term. AB - To elucidate diurnal variations in eye movement and fetal heart rate (FHR) variability in the term fetus, we observed these two parameters continuously for 24 h, using real-time ultrasound and Doppler cardiotocograph, respectively. Studied were five uncomplicated fetuses at term. The time series data of the presence and absence of eye movement and mean FHR value for each 1 min were analyzed using the maximum entropy method (MEM) and subsequent nonlinear least squares fitting. According to the power value of eye movement, all five cases were classified into two groups: three cases in the large power group and two cases in the small power group. The acrophases of eye movement and FHR variability in the large power group were close, thereby implying the existence of a diurnal rhythm in both these parameters and also that they are synchronized. In the small power group, the acrophases were separated. The synchronization of eye movement and FHR variability in the large power group suggests that these phenomena are governed by a common central mechanism related to diurnal rhythm generation. PMID- 11408102 TI - Upper limb deficiencies in Swedish children--a comparison between a population based and a clinic-based register. AB - OBJECTIVE: To validate information in the Swedish Register for Congenital Malformations (SRCM). METHODS: A comparison was made with a clinic-based register kept at the Limb Deficiency and Arm Prosthesis Centre (LDAPC). The report frequency and the quality of the information in SRCM were analysed. Cases were classified according to a detailed, clinically relevant classification, the ISO 8548-1:89 method. OUTCOME MEASURES: The completeness of SRCM was first estimated. The Kappa statistic was then used to assess the agreement between the two registers regarding individual categories and across all categories. RESULTS: For the period 1973-1987, we found 125 cases of upper limb reduction deficiencies (ULRD) in the clinic-based register, of which 117 was found in the national register. The completeness of SRCM was thus estimated to be 94% (95% confidence interval 89-98%). The inter-register agreement varied from almost perfect agreement in laterality of deficiency (Kappa 0.98) to substantial agreement in type and level of deficiency (Kappa 0.72-0.79). For specific levels of transverse deficiency, however, the agreement varied between -0.05 and 0.66. CONCLUSIONS: The results indicate that SRCM, with its calculated underestimation of 6%, can be used for studying the prevalence of ULRD in Sweden. However, as SRCM is a surveillance register, the quality of some information seems to be low, making detailed description of cases difficult. Use of the population register data for clinical purposes could therefore result in lower validity. Additional information and follow-up of specific cases are therefore recommended. PMID- 11408103 TI - Excessive infant crying: often not confined to the first 3 months of age. AB - Excessive crying/fussing in infancy may account for serious problems in mother infant interactions but is believed to be self-limiting around the age of 3 months. A random digit dialling telephone survey weighted by sex, age, and population density and yielding 662 children aged 9 to <36 months was used to estimate prevalence and the proportion of cases where this problem extends beyond the third month postpartum. Excessive infant crying/fussing during the first 3 months of age was found in 21.0% (95% CI 17.9-24.1) and persisted for longer in 39.6% (95% CI 31.5-47.7) of these. Professionals consulting parents on crying problems should take into account its possible persistence for longer periods of time. PMID- 11408104 TI - International Child Care Practices Study: infant sleep position and parental smoking. AB - The International Child Care Practices Study (ICCPS) collected descriptive data from 21 centres in 17 countries. In this report, data are presented on the key sudden infant death syndrome (SIDS) risk factors of infant sleep position and parental smoking. METHODS: Using a standardised protocol, parents of infants were surveyed at birth by interview, and at 3 months of age mainly by postal questionnaire. Data entry and descriptive analysis were undertaken with Epi Info. Centres were grouped according to geographic location. Also indicated was the level of SIDS awareness in the community, i.e. whether any campaigns or messages to "reduce the risks of SIDS" were available at the time of the survey. RESULTS: Birth interview data were available for 5488 individual families, and 4656 (85%) returned questionnaires at 3 months. Overall, 52.5% (95% CI, 43.5-61.3) of infants were placed in the supine or back sleep position, 37.8% (95% CI, 30.6 45.7) in the side position and 13% (95% CI, 9.3-17.9) in the prone or front position. The prevalence of supine sleeping ranged from 14% (Santiago, Chile) to 89% (Tokyo/Yokohama, Japan). The prevalence of reported smoking was between 0% and 34% for mothers and 10% and 64% for fathers. In samples from Western countries, there was a trend for rates of maternal smoking to be similar to paternal smoking, whereas in other samples, low rates of maternal smoking contrasted with high rates of paternal smoking. CONCLUSIONS: These data highlight considerable variations and interesting patterns in the prevalence of these two important SIDS risk factors in these diverse samples. Impressive differences in rates of smoking within and between these communities may reflect different stages of the inevitable progression of the smoking epidemic, but may also provide some encouragement that reduction in smoking rates, in both mothers and fathers, is possible. PMID- 11408105 TI - Simultaneous measurement of antibodies to three HIV-1 antigens in newborn dried blood-spot specimens using a multiplexed microsphere-based immunoassay. AB - We developed a fluorescent immunoassay to simultaneously measure antibodies to three HIV-1 antigens from newborn dried blood-spot specimens. The multiplexed assay uses fluorescent microspheres and a flow analyzer. The procedure is sensitive, precise and accurate, and can be expanded to simultaneously measure additional multiple analytes from a single specimen. PMID- 11408106 TI - Diurnal changes in the power spectral characteristics of eye movements and heart rate variability in the human fetus at term. AB - It is well known that 1/f characteristics in power spectral patterns exist in various biological factors including heart rate variability. In the present study, we tried to elucidate the diurnal variation in spectral properties of eye movement and heart rate variability in the human fetus at term, via continuous 24 h observation of both these parameters. Studied were five uncomplicated fetuses at term. We observed eye movement and fetal heart rate (FHR) with real-time ultrasound and Doppler cardiotocograph, respectively, and analyzed the diurnal change in spectral properties, using the maximum entropy method. In four of five cases, the slope values of power spectra for both eye movement frequency and FHR, ranging approximately between 0.5 and 1.8, indicated diurnal variation, where the slopes tended to have high values during the day and low values at night. These findings suggest that, in the human fetus at term, eye movement and FHR are under the control of a common central mechanism, and this center changes its complexity as seen through diurnal rhythm. PMID- 11408107 TI - Gender-related differences in rectal temperature in human neonates. AB - The aim of this study was to examine the gender-related differences in human neonates' body temperature. Rectal temperatures of 101 newborns (52 girls and 49 boys) were measured using a calibrated glass-mercury thermometer five times during their first 5 days of life. Results show that the temperature of males, averaged over 5 days, was significantly lower (37.068 degrees C) than that of females (37.168 degrees C). This result suggests that gender-dependent differences in baseline body temperature are present at birth. PMID- 11408108 TI - Maternal smoking and reduced duration of breastfeeding: a review of possible mechanisms. AB - AIM: To systematically review the possible mechanisms by which maternal smoking affects lactation. METHODS: Databases (Medline, CINAHL, Current Contents, Psychinfo, Sociological Abstracts and the Cochrane Library) were searched for smoking and breastfeeding or infant feeding. The Journal of Human Lactation and Birth were hand-searched, searches were also conducted at NMAA's Lactation Resource Centre and references cited were located where appropriate. Articles were divided into the various ways that smoking could affect breastfeeding and were tabulated. RESULTS: Most studies were conducted on small samples of animals or humans; the majority were prior to 1985. Most animal studies exposed the animals to much greater levels of nicotine than those to which humans are exposed. Most studies did not examine if breastfeeding behaviour was similar in smokers and non-smokers, and did not consider that any physiological difference found could be the result of poor lactation practices, rather than the cause of poor lactation in smokers. The definition of breastfeeding infants was also problematic in many studies. The effect of smoking on oxytocin in women was only examined in one study, and no effect was found, yet a negative effect of smoking on oxytocin release is reported in the breastfeeding literature. CONCLUSION: Although there is consistent evidence that women who smoke breastfeed their infants for a shorter duration than non-smokers, the evidence for a physiological mechanism is not strong. PMID- 11408109 TI - X-autosome translocation and low fertility in a family of crossbred cattle. AB - An investigation was carried out on a family of Limousin-Jersey crossbreds exhibiting low fertility in the females, to determine the impact of a previously identified X-autosome translocation (X-AT) on the reproductive performance of the carrier cows. Three of the identified translocation carriers, including a cow and two of her daughters, were maintained at our University Research Station and artificially inseminated periodically with semen from different bulls of known fertility. Attempts to breed the X-AT carriers resulted in high rates of return to estrus between days 28 and 60, abortions between days 121 and 235 after insemination, and a total of 13 live births including 4 translocation carrier calves. Results of superovulation and embryo retrieval trials on X-AT carriers revealed significantly higher proportions of unfertilized and uncleaved ova and abnormal embryos compared to those from normal cows, and no pregnancy in the recipients transferred with morphologically normal blastocysts from X-AT carriers. While the higher rates of failed fertilization and cleavage, abnormal embryos and return to estrus in X-AT carriers could be attributed to chromosome imbalance expected in their gametes, the relatively high prevalence of abortion (late in gestation) was unexpected. Our observations on the fetuses expelled by X AT carriers after 5 months of gestation indicated that a majority (three out of four) of these fetuses were products of abnormal (3:1) segregation in meiosis I and that these chromosomally unbalanced (hyperdiploid) conceptuses were able to survive early embryogenesis and fetal life up to the end of the second trimester. We hypothesize that their relatively long in utero life and the absence of any overt birth defects may be attributable to the type of chromosomes over represented in these fetuses and that their eventual expulsion may have been the result of selection against the clonal population of cells in which the altered X carrying a segment of chromosome 23 (Xp(+)), remained inactive. PMID- 11408110 TI - Bovine nuclear transfer embryo development using cells derived from a cloned fetus. AB - Many different cell types have been used to generate nuclear transfer embryos and fetuses. However, little is known about the potential of fibroblasts derived from a nuclear transfer fetus as donor cells for nuclear transfer. The ability of cloned fetuses or animals to be cloned themselves is of great interest in determining whether successive generations of clones remain normal or accumulate genetic or phenotypic abnormalities. We generated a bovine fibroblast cell line from a cloned fetus, that continued to divide beyond 120 days (94 doublings,18 passages) in continuous culture. As long-term survival of cells in culture is a desirable characteristic for use in transgenic cell production, passage 2 and 18 cells were compared as donor cells for nuclear transfer (NT). When cells from passage 2 (2 weeks in culture) and passage 18 (4 months in culture) were used for nuclear transfer, there was no significant difference in development rate to blastocyst (35.4 versus 44.6%, P=0.07). A greater proportion of late passage cells were in G0/G1 whether under serum-fed (64 versus 56%, P<0.01) or serum starved (95 versus 88%, P<0.01) culture conditions. Following embryo transfer, equivalent day 30 pregnancy rates were observed for each group (P 2: 2/19 versus P 18: 2/13). A slightly retarded fetus was surgically removed at day 56 and the remaining three fetuses died in utero by day 60 of gestation. Our results show that fibroblast cells derived from regenerated cloned fetuses are capable of both in vitro and in vivo development. The longevity of this regenerated cell line would allow more time for genetic manipulations and then to identify stable transfected cells prior to their use as NT donor cells. Although no live fetuses were produced in this study the results provide encouraging data to show that a cloned fetus can itself be recloned to produce another identical cloned fetus. Further studies on this and other recloned fetuses are necessary to determine whether the failure to produce live offspring was a result of inadequate sample size or due to the cell type selected. PMID- 11408111 TI - Incidence of premature estrus in lactating dairy cows and conception rates to standing estrus or fixed-time inseminations after synchronization using GnRH and PGF(2alpha). AB - Fixed-time AI (TAI) after GnRH-PGF(2alpha)-GnRH treatment is a method to achieve pregnancies in dairy herds without estrous detection. However, cows that fail to respond to the initial GnRH may have compromised TAI conception rates due to asynchronous ovarian response. This study documented the percentage of GnRH treated Holstein cows (n=345) in two herds that displayed estrus at an inopportune time for optimum TAI conception rate (< or =48h post-PGF(2alpha); premature estrus (PE)) and compared conception rates of two TAI protocols in cows that did not display PE. At biweekly herd health exams, cows diagnosed as not pregnant to a previous AI and cows >80 days postpartum with no AI were treated with 100 microg GnRH (day -7) and 25mg PGF(2alpha) (day 0). Cows detected in PE by twice-daily visual observation from day -7 to day 2 were bred by AI 8-12h later. Cows not detected in PE were randomly assigned by parity, body condition score, and postpartum interval to receive either: (1) 100microg GnRH at 48h after PGF(2alpha) and TAI 16 to 18h later (Ovsynch); or (2) TAI at 72h post-PGF(2alpha) and a concurrent 100 microg GnRH injection to those cows not detected in estrus between 48 and 72h post-PGF(2alpha) (modified Ovsynch (MOV)). All hormone injections were im. Twenty percent (68/345) of the cows were detected in estrus before 48 after PGF(2alpha), of which 5% (17/345) were detected in estrus before PGF(2alpha) (< or =day 0). Herd influenced the percentage of cows in the PE group (herd A versus herd B; 25% versus 14%; P<0.05). Conception rates were not affected by treatment (PE versus Ovsynch versus MOV; 32% (21/65) versus 30% (37/125) versus 32% (47/145); P>0.10). However, within MOV-treated cows, conception rates were greater (P<0.05) in cows detected in estrus (46% (23/50)) compared with cows not detected in estrus (25% (24/95)). In conclusion, 20% of GnRH-treated cows displayed PE and necessitates estrous detection during this period if maximal pregnancy rates are to be achieved. Although additional estrous detection is required compared to Ovsynch, reduced cow handling and hormone usage, efficient use of expensive semen through greater conception rates in cows detected in estrus, and comparable TAI conception rates, suggests the MOV protocol may be a cost effective alternative to Ovsynch in many dairy herd reproductive management programs. PMID- 11408112 TI - Relationship between follicle size and ultrasound-guided transvaginal oocyte recovery. AB - We evaluated the relationship between follicle size and oocyte recovery (OR) using ultrasound-guided follicle aspiration. Thirty Holstein cows were subjected to OR without gonadotrophic therapy. Oocytes were recovered two to four times from each cow in a total of 67 aspiration sessions. Ovarian follicles with diameters < or =4 mm and >4 mm were aspirated in separated groups. Recovered oocytes from each group were kept separate and submitted to in vitro maturation, fertilization, and culture to the blastocyst stage. A total of 430 follicles were aspirated, of which 154 (35.8%) were from follicles >4 mm and 276 (64.2%) were from follicles < or =4 mm. Seventy-seven oocytes (50%) were recovered from follicles >4 mm and 200 (72.2%) were from follicles < or =4 mm. Nineteen blastocysts were obtained from follicles >4 mm, whereas 45 blastocysts were obtained from follicles < or =4 mm. Recovery rate was greater (P<0.01) in follicles < or =4 mm. Oocyte quality, cleavage rate and blastocyst development did not differ between different follicle sizes. Routine aspiration of small follicles (< or =4 mm) could increase the number of oocytes available for in vitro development. PMID- 11408113 TI - Receptors for insulin-like growth factor-I and tumor necrosis factor-alpha are hormonally regulated in bovine granulosa and thecal cells. AB - Mastitis induces release of tumor necrosis factor-alpha (TNFalpha) and has been linked with reduced reproductive performance. To further elucidate the role and mechanism of action of TNFalpha on ovarian cells, the effect of TNFalpha on insulin-like growth factor-I (IGF-I)-induced steroidogenesis and IGF-I binding sites in granulosa and thecal cells as well as the hormonal regulation of TNFalpha receptors were evaluated. Granulosa and thecal cells were obtained from small (1-5mm) and large (> or =8mm) bovine ovarian follicles, respectively, and cultured for 3-4 days. During the last 2 days of culture, cells were treated with various hormones and steroid production and specific binding of 125I-IGF-I and 125I-TNFalpha was determined. Two-day treatment with 30 ng/ml of TNFalpha decreased (P<0.05) IGF-I-induced estradiol production by granulosa cells and IGF I-induced androstenedione production by thecal cells. Two-day treatment with 10 and 30ng/ml of TNFalpha decreased (P<0.05) specific binding of 125I-IGF-I to thecal cells, but had no effect on specific binding of 125I-IGF-I to granulosa cells, or on specific binding of 125I-IGF-II to thecal cells. TNFalpha did not compete for 125I-IGF-I binding to granulosa or thecal cells whereas unlabeled IGF I suppressed 125I-IGF-I binding. Insulin inhibited (P<0.10) whereas FSH had no effect on the number of specific 125I-TNFalpha binding sites in granulosa cells. In contrast, LH increased (P<0.10) whereas insulin had no effect on specific 125I TNFalpha binding sites in thecal cells. These results suggest that IGF-I and TNFalpha receptors in granulosa and thecal cells are regulated by hormones differentially. PMID- 11408114 TI - Sperm function and production of bovine embryos in vitro after swim-up with different calcium and caffeine concentration. AB - Frozen semen from bulls was used in artificial insemination programs was submitted to swim-up in Sperm Talp media containing different calcium (1.8, 2.6, 3.6 mM) or caffeine (2.5, 5.0, 7.5 microM) concentrations. The following sperm variables were evaluated: sperm recovery, motility, vigor, morphology, alterations in the pattern of capacitation by chlortetracycline (CTC) staining, and alterations in lysophosphatidyl choline (LPC)-induced acrosome reaction (AR). Sperm obtained from swim-up under different conditions were also tested for in vitro embryo production. No significant differences in the variables motility, vigor, morphology, and LPC-induced AR were observed among the treatments. However, the use of caffeine resulted in greater frequency of sperm with the capacitated pattern by CTC staining, compared to controls without caffeine. The greatest frequency of capacitated sperm (53%) was observed with 7.5 microM caffeine. Different calcium and caffeine concentrations in swim-up resulted in no significant differences in the cleavage rate and embryo development. In summary, micromolar concentrations of caffeine in Sperm Talp may stimulate sperm capacitation. PMID- 11408115 TI - Effects of cooling and warming conditions on post-thawed motility and fertility of cryopreserved buffalo spermatozoa. AB - The principal objective of this study was to derive an improved procedure for cryopreservation of swamp buffalo (Bubalus bubalis) spermatozoa. Experiments were conducted to determine effects of cooling rate, intermediate plunge temperature and warming rate on motility and acrosome integrity of spermatozoa. Spermatozoa were obtained from three bulls (three ejaculates/bull) and were subjected to nine cooling conditions before being frozen in liquid nitrogen: cooling at 10, 20, or 30 degrees C/min each to -40, -80, or -120 degrees C before being plunged into liquid nitrogen. The spermatozoa frozen under a given condition were then thawed either at 1000 or 200 degrees C/min. Cooling rate, intermediate temperature and warming rate significantly affected survival of spermatozoa obtained from the three bulls. Cooling spermatozoa from 4 to -120 degrees C either at 20 or 30 degrees C/min yielded better progressive motility compared to other cooling conditions (50 versus 30%). Rapid warming was superior to slow warming. In an additional study, motility and fertility of spermatozoa frozen after being cooled to -120 degrees C at 20 degrees C and 30 degrees C/min and those frozen by a standard protocol used routinely for semen processing were assessed. Progressive motility of cryopreserved spermatozoa cooled at 20 degrees C and 30 degrees C/min was 40%, while that of spermatozoa cryopreserved using a standard protocol was 25%. A total of 178 buffalo cows were inseminated with cryopreserved spermatozoa obtained from one bull, and their pregnancy status was assessed 60 days later by rectal palpation. Out of the 60, 26 (43%) and 23 of 58 (40%) cows inseminated with sperm cooled at 20 and 30 degrees C/min, respectively, became pregnant, whereas 17 of 60 (28%) cows inseminated with sperm frozen by a standard protocol became pregnant. This study demonstrates that an effective cryopreservation procedure for buffalo spermatozoa can be derived by systematic examination of various cryobiological factors. PMID- 11408116 TI - Circulating gonadotrophins and follicular dynamics in anestrous ewes after treatment with estradiol-17beta. AB - Plasma FSH, LH, estradiol (E2) and progesterone (P4) profiles and patterns of follicular growth and regression by ultrasonography were determined after E2 treatment (1 microg/kg) in anestrous ewes. Fifteen ewes were treated with one (group I, n=7) or two (group II, n=4) i.m. injections of E2 with a 24h interval, or two oil injections with a 24h interval (group C, n=4). Blood samples for E2, P4, FSH and LH determinations were collected daily 4 days before the initiation of the treatment (day 0), when bleeding increased to every 2h starting 2h before treatment until 56h after the first injection and from then on every 6h until day 8, and twice per day till the end of the experiment (day 9). During the experimental period (days -4 to 9), transrectal ultrasonic examinations were carried out daily using a 7.5 MHz linear array probe. Number and size of follicles > or =3mm in diameter were recorded. No estrous was detected before, during or after treatment. LH and FSH surges were observed 10-18h after the first E2 injection. The second E2 injection stimulated another release of LH but no surges. E2 inhibited FSH levels before the surge and the second E2 injection induced a longer inhibition. No ovulation was detected by ultrasonography during the experimental period and P4 levels remained low (<0.7 nmol/l) before, during and after the treatment in all ewes. There was an effect of E2 treatment on the diameter of the largest follicle, a decrease could be observed 3 days after the first injection in both ewes of groups I and II. The E2-treated groups had a higher frequency of ewes showing wave emergence on day 3 (day 1.5+/-1,2.4+/-0.4 and 2.5+/-0.5 for control, groups I and II). LH and FSH surges were observed after E2 treatment, but were not able to provoke ovulation neither luteinization. In contrast, the treatment was associated with the regression of the largest follicle and with emergence of a new follicular wave on day 3. PMID- 11408117 TI - The influence of parity order and body condition and serum hormones on weaning-to estrus interval of sows. AB - This study was performed to investigate the weaning-to-estrus interval (WEI) in primiparous and multiparous sows in relation to their weight and back-fat thickness changes and serum hormone imbalance (insulin, growth hormone, and cortisol) at the end of gestation and during lactation. Ten primiparous and ten multiparous Camborough sows, fourth to seventh parity, were used in this experiment. During gestation, daily food intake was 2.4 kg (sow commercial diet: 2.96 Mcal/kg, 16% crude protein) and during lactation all sows were fed on a wet commercial diet (3.34 Mcal/kg, 17% crude protein) ad libitum, three times per day. Blood samples were collected and back-fat thickness at the P(2) site were recorded at 6 days before and 2, 7, 14, 21 and 25 days after farrowing. Body weight was recorded on the same dates, except the date before farrowing. The WEI was also recorded. The average daily feed intake was different (P<0.05) between primiparous and multiparous sows during lactation (5.23 versus 5.72kg per day, respectively). There was a difference (P<0.05) between primiparous and multiparous sows in total percentage of back-fat thickness loss from the end of gestation until weaning (-20.18 and -9.03%, respectively). The total percentage of weight loss during lactation was slightly greater (P>0.05) in multiparous than primiparous sows. Weaning-to-estrus interval was greater (P<0.05) in the primiparous group when compared with the multiparous group (5.55 and 4.22 days, respectively). No differences were found in insulin, growth hormone (GH), and cortisol concentrations between parity groups, except on the 21st day of lactation, when GH was greater in primiparous sows. There was no correlation between percentage of total weight loss and WEI, or percentage of back-fat thickness loss (total or by periods) and WEI. There were positive correlations between GH serum concentration on the 14th and 21st days and the percentage of weight loss in the third week of lactation (r=0.46, P<0.04 and r=0.52, P<0.02, respectively), and between GH concentration on the 21st and on weaning days and WEI (r=0.54, P<0.02 and r=0.42, P<0.06, respectively). Our results indicate that the hormone change (imbalance) during lactation, mainly GH, seems to be a better parameter to explain the difference in WEI between primiparous and multiparous sows than change in body condition. PMID- 11408118 TI - Cryopreservation of epididymal dog sperm. AB - A method of cryopreservation was developed for sperm salvaged from the cauda epididymis and vas deferens of domestic dog testes. Four modifications of the glycerol concentration of a buffer used for cryopreservation of dog ejaculates and two freezing rates were assessed for their effect upon post-thaw spermatozoal motility and morphology. There was no statistical difference between the four glycerol concentrations or the two freezing rates and the buffer containing 6% glycerol and the freezing rate provided by 0.5 ml straws was chosen for further study. This method resulted in a significant reduction in the percentage of live spermatozoa detected with Hoechst staining and a reduction in the percentage of capacitated spermatozoa after freeze-thawing. However, there was no difference in the ability of frozen-thawed spermatozoa to penetrate homologous oocytes. This study demonstrates that cryopreservation of epididymal canine sperm can be performed using methods similar to those established for ejaculates of the same species, and that despite some damage, spermatozoa retain their functional ability. PMID- 11408119 TI - Sodium cloprostenol administered at a continuous low dosage induces polydipsia and suppresses luteal function in early dioestrous bitches. AB - The aim of this study was to determine whether sodium cloprostenol administered at a continuous low dosage induced luteolysis and polydipsia in early dioestrous bitches. Sodium cloprostenol was administered subcutaneously to greyhounds at doses of 4.04-5.19 microg/kg/day (treated group, n=5) or 0 microg/kg/day (control group, n=5) delivered by mini-osmotic pumps for 7 days. The treated bitches and two of the control bitches were in early dioestrus (Days 5-14, and 6 and 10, respectively) when the mini-osmotic pump was inserted (Day 0). Concentrations of plasmatic progesterone were measured in dioestrous bitches each day from Day -2 to 7, and then weekly until Day 90. Daily intake of water was ascertained in all bitches from Day -2 until Day 10, and their weight was measured on Days -2, 6 and 13. Biochemical analyses on plasma for concentrations of urea and glucose, and urinalyses were performed on all bitches before (Day -1), during (Day 4) and after treatment (Day 10). Concentrations of plasmatic progesterone declined dramatically and rapidly in treated bitches after Day 0 to <2.9 ng/ml but were not similarly affected in the dioestrous control bitches. However, in three of five treated bitches, concentrations of plasmatic progesterone increased to >1 ng/ml in the period from Day 10 to 90 indicating that luteolysis was incomplete. All treated bitches were polydipsic (intake of water >100 ml/kg/day) for 2-6 days during the period of treatment, and for 0-2 days immediately after treatment (Days 7 and 8). One control bitch was polydipsic on Days -2, -1 and 0. The treated bitches were also polyuric since they were hyposthenuric (<1.007, n=4) or isothenuric (1.010, n=1) on Day 4, their weight did not increase and no gastrointestinal or respiratory effects were observed. The control bitches were always hypersthenuric when measured during and after treatment (>1.021). Biochemical analyses of plasma and other data obtained from urinalyses did not reveal any differences between groups. This study indicated that sodium cloprostenol administered at a continuous low dosage induced polydipsia and suppressed luteal function in early dioestrous bitches. PMID- 11408120 TI - Cortisol concentrations in the perinatal and weaning periods of alpacas. AB - Cortisol concentrations were determined during the perinatal and weaning periods in alpacas. Fifty males and 50 females were chosen at random (25 at each period) for blood collection on day of parturition, 3 and 5 days after birth. For the weaning period, blood samples were collected 2 days before, on the day of weaning (0), and at days 3 and 5 after weaning. Cortisol was determined using an ELISA protocol validated for the alpaca. There was no difference (P>0.05) in cortisol concentrations in both periods, perinatal and weaning between male and female crias. Cortisol was elevated at day of parturition (125.8 ng/ml) and then decreased to 27.2 ng/ml at 3 days after birth (P<0.05). Conversely, cortisol was 32.5 ng/ml 2 days before weaning and then increased at day 3 to 64.7 ng/ml (P<0.05), but then decreased to 29.4 ng/ml at day 5 after weaning. Cortisol is elevated in new-born alpacas and then decreases at the third and fifth after birth. Concentrations of cortisol are also increased after weaning and then decreased 5 days after weaning. PMID- 11408123 TI - Translimbal approach for intravitreal injection in endophthalmitis after phacoemulsification. AB - We describe a corneal limbal technique of intravitreal injection for use in cases in which it is difficult to confirm the position of the needle. Using this translimbal approach, the needle's position is easily identified and the intravitreal injection can be given using topical anesthesia. This technique is an option in eyes with a cloudy cornea or a large iridectomy. PMID- 11408124 TI - Teaching continuous curvilinear capsulorhexis using a postmortem pig eye with simulated cataract(2)(2). AB - We modified an inexpensive, easily prepared model using a postmortem pig eye for training ophthalmology residents to perform continuous curvilinear capsulorhexis. To reduce the tension and elasticity of the porcine anterior lens capsule, 0.05 mL of formalin mixed with hydroxyethylcellulose or a viscoelastic material is injected into the anterior chamber via the corneal limbus to fix the surface of the central anterior lens capsule of the pig eye in situ. One or 2 minutes after the injection, only the anterior lens capsule is fixed and corneal transparency is maintained. The reduction in tension and elasticity of the anterior lens capsule caused by its fixation increases the resemblance of the simulated cataract to the human cataract. PMID- 11408125 TI - Changes in posterior capsule opacification after poly(methyl methacrylate), silicone, and acrylic intraocular lens implantation. AB - PURPOSE: To prospectively evaluate the progression of posterior capsule opacification (PCO) after poly(methyl methacrylate) (PMMA), silicone, and acrylic intraocular lens (IOL) implantation. SETTING: Hayashi Eye Hospital, Fukuoka, Japan. METHODS: Three hundred eyes of 300 patients scheduled to have IOL implantation were initially randomized into 3 groups based on IOL type: PMMA, silicone, or acrylic. Of the 300 eyes, 269 completed the follow-up. The PCO density in these eyes was measured 1 week and 3, 6, 12, 18, and 24 months postoperatively using special computer software developed for the Scheimpflug videophotography system. Visual acuity and the incidence of neodymium:YAG (Nd:YAG) laser capsulotomy were also examined. RESULTS: Three months postoperatively and later, the mean PCO value in the PMMA group increased significantly (P <.0001); the increase in the silicone and acrylic groups was not significant. The PCO value in the PMMA group was significantly greater than in the silicone or acrylic group (P <.0001). The PCO value in the acrylic group was slightly less than in the silicone group at 18 and 24 months, but the difference was marginal. The survival rate not requiring Nd:YAG capsulotomy was least in the PMMA group, followed by the silicone and acrylic groups in that order (P <.0001). The mean logMAR visual acuity in the PMMA group increased postoperatively and was worse than in the silicone or acrylic group. CONCLUSIONS: The degree of PCO after PMMA IOL implantation progressed significantly with time, while the progression after silicone and acrylic IOL implantation was slight. Therefore, PCO in eyes with a PMMA IOL was significantly more extensive than in those with a silicone or acrylic IOL and resulted in marked impairment of visual acuity. PMID- 11408126 TI - Posterior continuous curvilinear capsulorhexis with hydrogel and silicone intraocular lens implantation: development of capsulorhexis size and capsule opacification. AB - PURPOSE: To evaluate the influence of primary posterior continuous curvilinear capsulorhexis (PCCC) on capsule opacification development and capsular bag changes within the first year after cataract surgery with 2 intraocular lenses (IOLs) of comparable design but different material. SETTING: Department of Ophthalmology, University of Vienna, Medical School, Vienna, Austria. METHODS: Thirty-seven patients with age-related cataract had bilateral small incision cataract surgery with a PCCC performed after capsular tension ring insertion. One eye was randomly assigned to receive a hydrogel IOL and the other eye, a silicone IOL. Standardized digital retroillumination photographs were taken 1 day, 1 week, and 1, 3, 6, and 12 months after surgery to evaluate changes in the dimensions of the anterior and posterior capsulorhexis opening area and the presence of anterior and posterior capsule opacification. RESULTS: The area of the anterior continuous curvilinear capsulorhexis (ACCC) opening was significantly reduced during the first 6 postoperative months. The shrinkage was more pronounced (-25%) in the silicone IOL group than in the hydrogel IOL group. Ten percent of eyes with a silicone IOL had marked shrinkage of the ACCC. The area of the PCCC did not change in eyes with a hydrogel IOL but was larger (+20%) in eyes with a silicone IOL. Anterior ongrowth was observed in 60% in the hydrogel group and in no eye in the silicone group. Anterior capsule fibrosis was observed in 90% in the silicone group and in 20% in the hydrogel group. Total closure of the PCCC was not observed within the first year, but posterior ongrowth was observed in 40% in the hydrogel group and 10% in the silicone group. CONCLUSIONS: Anterior capsulorhexis shrinkage with concomitant posterior capsulorhexis enlargement was observed in eyes with a silicone IOL. The hydrogel IOL induced more ongrowth on the anterior and posterior IOL surfaces, whereas the silicone IOL induced more anterior capsule fibrosis. Total closure of the PCCC was not observed within the first year after surgery. PMID- 11408127 TI - Collamer intraocular lens: clinical results from the US FDA core study. AB - PURPOSE: To assess the safety and efficacy of the Staar Collamer intraocular lens (IOL) 1 year postoperatively. SETTING: Fifteen private ophthalmology practices geographically distributed throughout the United States. METHODS: Six hundred eighty-six cases with a mean age of 72.1 years were enrolled in a 2-phase U.S. Food and Drug Administration (FDA) clinical study of the Collamer IOL implanted during phacoemulsification for cataract removal. The 12 month follow-up comprised data from 502 cases. RESULTS: Twelve months postoperatively, 96.2% of all cases and 95.6% without preexisting pathology achieved 20/40 or better best corrected visual acuity. Both results were better than FDA Grid values. No persistent sight threatening complications, IOL dislocations, or IOL removals were reported. CONCLUSIONS: Results show that the Collamer IOL is safe and effective for use in small incision cataract surgery. PMID- 11408128 TI - Piggyback foldable intraocular lens implantation in patients with microphthalmos. AB - PURPOSE: To evaluate the clinical results of phacoemulsification and implantation of 2 foldable acrylic intraocular lenses (IOLs) in microphthalmic eyes. SETTING: University of Tokyo School of Medicine, Tokyo, Eguchi Eye Hospital, Hakodate, and Osaka Rosai Hospital, Osaka, Japan. METHODS: This study comprised 5 eyes of 3 patients whose manifest spherical equivalent was +10.5 to +18.0 diopters (D) and axial length was 15.79 to 16.82 mm. After phacoemulsification, 2 foldable acrylic IOLs with a mean power of +43.0 D +/- 9.5 (SD) (range +32.0 to +55.0 D) were implanted in the capsular bag. RESULTS: There were no significant intraoperative complications, and there was significant improvement in uncorrected and best corrected visual acuities after surgery. The spherical equivalent decreased significantly; however, the postoperative refraction was considerably more hyperopic than predicted in all eyes. Except for posterior synechias in 2 eyes and slight interlenticular opacification in 2 eyes, no major postoperative complications occurred during the mean follow-up of 14.8 months. CONCLUSIONS: Implanting 2 piggyback IOLs was beneficial in eyes with an extremely short axial length. However, the current system of lens power calculation markedly underestimates the required lens power, resulting in a hyperopic refractive error after surgery. The benefits of in-the-bag placement of 2 foldable lenses are questionable. PMID- 11408129 TI - Surface abnormalities on hydrophilic acrylic intraocular lenses implanted by an injector. AB - PURPOSE: To describe and evaluate abnormalities on the surface of hydrophilic acrylic intraocular lenses (IOLs) that were implanted using an injector. SETTING: Department of Ophthalmology, University of Graz, Graz, Austria. METHODS: This study comprised 70 eyes having phacoemulsification and implantation of an EasAcryl (n = 50) or Inject-A (n = 20) IOL. Both hydrophilic IOLs are biconvex and have a high water content (26%), plate-haptic design, and no angulation. The IOLs were folded in a cartridge under hydroxypropyl methylcellulose and implanted by an injector. RESULTS: Thirteen EasAcryl and 5 Inject-A IOLs had multiple white lines in the direction of the longitudinal axis on the posterior surface immediately after implantation. On 1 EasAcryl IOL, the white lines were separated by a clear central zone and were on the anterior surface. Another EasAcryl IOL had a 1.0 mm crack. CONCLUSIONS: The abnormalities on the posterior surface of high-water-content hydrophilic acrylic IOLs disappeared or were almost gone within 1 month postoperatively and do not appear to be a concern for surgeons. These abnormalities are likely the result of the IOLs' hydrophilicity and high water content and friction in the injector barrel. PMID- 11408130 TI - Lens epithelial cell outgrowth on 3 types of intraocular lenses. AB - PURPOSE: To compare the outgrowth of lens epithelial cells (LECs) on 3 types of intraocular lenses (IOLs) to determine the influence of lens material and lens design (optic edge) on this phenomenon. SETTING: Department of Ophthalmology, University of Vienna, Medical School, Vienna, Austria. METHOD: Ninety eyes scheduled for cataract surgery were included in a prospective comparative study. A standardized surgical procedure was performed by 1 experienced surgeon. Patients received 1 of 3 types of posterior chamber IOLs of similar design with a 6.0 mm optic and poly(methyl methacrylate) haptic: AcrySof (Alcon), HydroView (Bausch & Lomb), or Sensar (Allergan). Each IOL type was implanted in 30 eyes. Postoperative biomicroscopic examinations were performed 1, 3, 7, 30, 90, and 180 days and 1 year after surgery. Lens epithelial cells in each quadrant of the anterior lens surface were subjectively graded. The product with the highest density and the number of quadrants with this density were used to measure LEC outgrowth. RESULTS: Statistically significant differences (P <.05) were seen between the hydrophilic IOL and the 2 hydrophobic lenses from day 30 until the final examination. The HydroView lens had a higher number of LECs on its anterior surface than the AcrySof or Sensar IOL. There were no statistically significant differences between the 2 acrylic IOLs at any measurement. CONCLUSION: The findings suggest that lens surface properties have a greater influence on LEC outgrowth than lens design (ie, sharp optic edge). PMID- 11408131 TI - Aqueous flare induced by heparin-surface-modified poly(methyl methacrylate) and acrylic lenses implanted through the same-size incision in patients with diabetes. AB - PURPOSE: To compare the degree of blood-aqueous barrier (BAB) breakdown in eyes of diabetic patients after phacoemulsification and implantation of heparin surface-modified poly(methyl methacrylate) (PMMA) or soft hydrophobic acrylic intraocular lenses (IOLs) performed using the same technique with the same incision size to determine the influence of the IOLs on postoperative inflammation independent of other surgical factors. SETTING: Department of Ophthalmology, University of Paris XIII, Bobigny, France. METHODS: In a prospective study, 44 eyes of 31 diabetic patients with or without mild to moderate diabetic retinopathy were randomly assigned to receive an HSM PMMA IOL (22 eyes) or a soft hydrophobic acrylic IOL (22 eyes) after standardized phacoemulsification surgery. Both types of IOLs had a 6.0 mm optic, were inserted unfolded, and were placed in the bag through a calibrated 6.0 mm superior scleral incision. Anterior chamber flare was measured preoperatively and 1, 7, 30, and 240 days postoperatively using the Kowa 500 laser flare meter. RESULTS: The mean flare value was higher on the first postoperative day in both groups. There were no statistically significant between-group differences in flare scores or clinical parameters preoperatively or at any postoperative visit. CONCLUSIONS: No significant difference was observed in inflammation between eyes having HSM PMMA IOL implantation or those having soft hydrophobic acrylic IOL implantation through the same-size incision. This indicates that hydrophobic acrylic and HSM PMMA materials induce the same degree of BAB breakdown after phacoemulsification in eyes of diabetic patients. PMID- 11408132 TI - Improved prediction of intraocular lens power using partial coherence interferometry. AB - PURPOSE: To evaluate the feasibility of using a new optical biometry technique, dual-beam partial coherence interferometry (PCI), to improve intraocular lens (IOL) power prediction in cataract surgery. SETTING: Department of Ophthalmology, Vienna General Hospital, and Institute of Medical Physics, University of Vienna, Vienna, Austria. METHODS: Preoperative axial length (AL) data obtained with PCI biometry and applanation ultrasound (US) biometry in 77 eyes of 51 patients was applied to 4 commonly used IOL power formulas. The refractive outcome and the mean absolute error (MAE) were calculated for each formula using both biometry methods. A linear multiple-regression model based on preoperative PCI biometry data was derived to predict the postoperative anterior chamber depth (ACD). The predictive power of this regression model was assessed by adding the predicted ACD to the SRK/T formula. Predicted residuals were calculated to evaluate the feasibility and stability of this modified IOL power formula. RESULTS: Using PCI instead of US biometry significantly improved the refractive outcome with all 4 IOL power formulas. The Holladay I and SRK/T formulas yielded an MAE of 0.44 diopter (D) using PCI AL data and 0.56 D and 0.57 D, respectively, using US biometry data. The SRK/T formula combined with the PCI regression model for postoperative ACD prediction performed slightly better (MAE 0.42 D) than the conventional SRK/T formula alone. Predicted residuals revealed an MAE of 0.46 D, proving the predictive performance of the new formula. CONCLUSIONS: Partial coherence interferometry biometry applied to several widely used IOL power formulas yielded significantly better IOL power prediction and therefore refractive outcome in cataract surgery than US biometry. Further improvement can be achieved by applying PCI to a modified SRK/T formula that predicts the postoperative ACD using PCI biometry data. PMID- 11408133 TI - Corneal surface changes after pars plana vitrectomy and scleral buckling surgery. AB - PURPOSE: To evaluate the changes in corneal shape after pars plana vitrectomy, encircling buckling surgery, and a combination of the 2 procedures. SETTING: Assaf Harofeh Medical Center, Zrifin, and Goldshlager Eye Institute, Tel Hashomer, Israel. METHODS: In a prospective clinical study, 61 eyes of 61 patients were assigned to 1 of 3 groups based on the type of procedure: pars plana vitrectomy, encircling buckle and pars plana vitrectomy, or encircling buckle. The eyes had corneal keratometry (TMS-1), visual acuity measurement, and videokeratography (TMS-1) preoperatively and 2 days, 1 week, and 1 and 3 months postoperatively. The topographic parameters statistically analyzed were the corneal surface cylinder, simulated keratometry, surface regularity index, surface asymmetry index, standard deviation of corneal power, average corneal power, and irregular astigmatism index. RESULTS: The measurements of all topographic indices were statistically significantly different from baseline measures 2 and 7 days postoperatively in all 3 groups. However, at 1 and 3 months, there was no significant difference from preoperatively in any index. Postoperative best corrected visual acuity was statistically significantly different from preoperatively. CONCLUSIONS: All corneal surface indices were significantly altered after all 3 types of surgery. Corneal surface measurements returned to preoperative values by 1 month postoperatively and remained stable until the final follow-up at 3 months. PMID- 11408134 TI - Topical versus sub-Tenon's anesthesia without sedation in cataract surgery. AB - PURPOSE: To compare pain control using topical anesthesia with that using sub Tenon's anesthesia for clear corneal phacoemulsification cataract surgery and foldable intraocular lens (IOL) implantation. SETTING: Departments of Ophthalmology, General Hospital Asklepeion Voulas and General Hospital of Athens, University of Athens, Athens, Greece. METHODS: One hundred consecutive patients scheduled for bilateral cataract surgery 1 to 2 months apart were prospectively randomized to receive topical anesthesia (100 eyes) or sub-Tenon's anesthesia (100 eyes). The randomization was stratified so that one half of first-eye surgeries and one half of second-eye surgeries were assigned to each anesthesia group, with each patient receiving each type of anesthesia once. All patients had clear corneal phacoemulsification with foldable IOL implantation. Patients were asked to rate their pain level on a 10-point scale for 4 periods: during the administration of the anesthetic agent, during surgery, immediately after surgery, and 24 hours postoperatively. The surgeon recorded his subjective assessment of ease of surgery and surgical complications using a standardized template. RESULTS: Eighty-one percent of patients who received topical anesthesia and 8% of patients who received sub-Tenon's anesthesia reported no pain during delivery of the anesthetic agent. The mean pain score was 0.19 +/- 0.39 (SD) in the topical group and 1.35 +/- 0.63 in the sub-Tenon's group. The difference between groups was statistically significant (P <.001). Seventy-two percent of patients in the topical anesthesia group and 86% in the sub-Tenon's anesthesia group reported no pain or slight discomfort during surgery (mean score 1.13 +/- 1.57 and 0.57 +/- 1.28, respectively) (P <.001). Ninety percent of topical anesthesia patients and 100% of sub-Tenon's anesthesia patients reported no pain or slight discomfort 30 minutes postoperatively (mean score 0.80 +/- 0.93 and 0.12 +/- 036, respectively) (P <.001). All patients in the topical anesthesia group and 77% in the sub-Tenon's group reported no pain 24 hours postoperatively (mean pain 0.00 +/- 0.00 and 0.23 +/- 0.40, respectively) (P <.001). Complications including prolonged akinesia of the globe, chemosis, and conjunctival hemorrhage occurred significantly more frequently in the sub-Tenon's than in the topical group (P <.001). CONCLUSIONS: Patients having cataract surgery under topical anesthesia had more intraoperative and postoperative discomfort than patients receiving sub-Tenon's anesthesia. However, patients having topical anesthesia reported less pain during its administration and had fewer complications. Both anesthesia methods provided high levels of pain control without additional sedation. PMID- 11408135 TI - Patient pain during different stages of phacoemulsification using topical anesthesia. AB - PURPOSE: To assess the pain experienced by patients during the different stages of phacoemulsification cataract surgery performed under topical anesthesia. SETTING: Ophthalmic teaching hospital, Dublin, Ireland. METHODS: This prospective study comprised 100 consecutive patients having phacoemulsification under topical anesthesia. Patients were asked to grade the pain they experienced during the different stages of the procedure using a visual analog pain scale from 0 to 10. The pain experienced during the procedure was compared with that experienced after the instillation of a drop of amethocaine. RESULTS: The overall mean pain score was 1.46. The highest mean pain score, which was during the phacoemulsification stage of the procedure, was not significantly more than the score for the administration of the topical anesthetic agent. The duration of surgery was not related to the level of pain during the procedure. CONCLUSIONS: Topical anesthesia was effective in phacoemulsification cataract surgery. Because the highest mean score was not significantly higher than that for the administration of the anesthetic agent, it is possible to counsel patients before surgery that the pain they experience during the procedure will be no worse than that during administration of the anesthetic drops. PMID- 11408136 TI - Effects of hand massage on anxiety in cataract surgery using local anesthesia. AB - PURPOSE: To evaluate the effects of hand massage on patient anxiety during cataract surgery. SETTING: Kangnam St. Mary's Hospital, Seoul, Korea. METHODS: This study comprised 59 patients having cataract surgery from December 11, 1996, to February 12, 1997. The patients were divided into those having a hand massage 5 minutes before surgery (experimental group, n = 29) and those not receiving a hand massage (control group, n = 30). Patients' anxiety levels were measured using the Visual Analog Scale and by assessing the systolic blood pressure, diastolic blood pressure, and pulse rate before and after the hand massage and 5 minutes before the end of surgery. Epinephrine, norepinephrine, cortisol, blood sugar levels, neutrophil, and lymphocyte percentages in white blood cells were also measured. RESULTS: After the hand massage, the psychological anxiety levels, systolic and diastolic blood pressures, and pulse rate were significantly lower than before the massage. The hand massage significantly decreased epinephrine and norepinephrine levels in the experimental group. Epinephrine, norepinephrine, and cortisol levels increased in the control group. The differences between groups were significant. There were no significant between-group differences in blood sugar levels or neutrophil and lymphocyte percentages in white blood cells. CONCLUSION: The findings indicate that hand massage decreases the psychological and physiological anxiety levels in patients having cataract surgery under local anesthesia. PMID- 11408137 TI - Customized ablation for asymmetrical corneal astigmatism. AB - PURPOSE: To evaluate laser in situ keratomileusis (LASIK) cases in which a customized ablation assisted by corneal topography was performed. SETTING: Instituto de la Vision, Buenos Aires, Argentina. METHODS: The study group comprised 23 cases with asymmetrical corneas; 17 had myopia and 6, hyperopia. Visual acuity, spherical equivalent (SE), and cylinder vectorial change measurements, along with keratometry and corneal topography, were performed in all cases preoperatively and postoperatively. RESULTS: In the myopic cases, the mean preoperative SE was -4.92 diopters (D) +/- 2.46 (SD); it was -0.38 +/- 0.66 D after a mean follow-up of 4.41 +/- 2.32 months. In the hyperopic cases, the preoperative SE was +3.54 +/- 1.57 D; it was +0.33 +/- 0.50 D after a mean follow up of 3.42 +/- 3.23 months. Cylinder vectorial change, visual acuity, and the number of visual acuity lines gained or lost were also evaluated in both groups. CONCLUSION: Although some features of our algorithm for corneal-topography assisted customized ablations should be modified, the results of our treatment of cases with asymmetrical astigmatism were encouraging. PMID- 11408138 TI - Clear lens phacoemulsification for correction of high myopia. AB - PURPOSE: To assess phacoemulsification and posterior chamber intraocular lens (IOL) implantation as an effective, safe, and predictable technique for the correction of high myopia. SETTING: University Eye Clinic of Verona, Verona, Italy. METHODS: A series of 25 eyes with myopia higher than -12.0 diopters (D) had clear lens extraction by phacoemulsification and IOL implantation in the capsular bag. The mean postoperative follow-up was 42.92 months +/- 3.76 (SD). RESULTS: No serious intraoperative complications occurred. Uncorrected visual acuity improved in all cases. The mean postoperative best corrected visual acuity improved by an average of 1 line. One case (4.0%) of postoperative retinal detachment (RD) occurred at 12 months. One case (4.0%) of biometric error (3.0 D) occurred. CONCLUSION: Clear lens extraction by phacoemulsification and IOL implantation in a series of highly myopic eyes was effective and had an acceptable predictability and a low rate of complications. Careful evaluation of the retinal periphery by indirect ophthalmoscopy is recommended to avoid postoperative RD. PMID- 11408139 TI - Laser in situ keratomileusis for residual myopia after radial keratotomy and photorefractive keratectomy. AB - PURPOSE: To evaluate the visual outcome, stability, and complications of laser in situ keratomileusis (LASIK) for residual myopia after radial keratotomy (RK) and photorefractive keratectomy (PRK). SETTING: Dr. Agarwal's Eye Hospital, Chennai, India. METHODS: Twenty eyes that had LASIK treatment for residual myopia after RK (10 eyes) or PRK (10 eyes) were retrospectively analyzed. Laser in situ keratomileusis was performed after a mean period of 24.3 months +/- 0.75 (SD) in the RK group and 22.0 +/- 1.07 months in the PRK group. RESULTS: At the last follow-up, the mean spherical equivalent was reduced from -6.05 +/- 1.98 diopters (D) to -1.26 +/- 0.32 D (P <.05) in the RK group and from -3.38 +/- 1.30 D to 0.55 +/- 0.40 D (P <.005) in the PRK group. The mean uncorrected visual acuity improved from 20/300 (range 20/600 to 20/200) to 20/40 (range 20/60 to 20/20) (P <.05) and from 20/200 (range 20/800 to 20/80) to 20/25 (range 20/40 to 20/20) (P <.05), respectively. Two eyes in the RK group and 3 in the PRK group gained 1 line of best corrected visual acuity, and 2 eyes in the RK group lost 1 line. No sight-threatening complications such as a free flap, corneal ectasia, or a retinal complication occurred. There was no statistically significant difference in corneal haze before and after LASIK. Two eyes in the RK group required repositioning of the flap because of irregular apposition to the stromal bed. CONCLUSION: Laser in situ keratomileusis was safe, effective, and stable in the treatment of residual myopia after RK and PRK. PMID- 11408140 TI - Effect of preoperative pupil measurements on glare, halos, and visual function after photoastigmatic refractive keratectomy. AB - PURPOSE: To prospectively assess the effect of preoperative variables such as pupil size on glare, halos, and visual function after photoastigmatic refractive keratectomy (PARK). SETTING: Department of Ophthalmology, Stanford University School of Medicine, Stanford, California, USA. METHODS: Ninety-three eyes had PARK for primary compound myopic astigmatism. Preoperative pupil diameters were measured under scotopic and photopic illuminance conditions. Postoperatively, patients were evaluated at 1, 3, 6, 9, 12, 18, and 24 months. A regression model was performed to evaluate the predictive value of assessing preoperative variables such as pupil diameter on the development of glare and halos, contrast sensitivity, and best spectacle-corrected visual acuity (BSCVA) under scotopic, photopic, and glare conditions. RESULTS: The greater magnitude loss of BSCVA under scotopic conditions in the early postoperative period as well as the slower recovery to preoperative levels in eyes with larger scotopic pupil diameters were not statistically significant (P >.05). An increase in symptoms of glare was related more to the attempted level of spherical equivalent (SE) correction than to the pupil size during the first 12 postoperative months (P <.01). The photoablation dimensions as determined by the attempted level of astigmatic correction may result in decreases in the glare BSCVA up to 12 months after PARK (P =.03). At the 2 year follow-up, pupil diameter under both scotopic and photopic illuminance conditions was not predictive of any of the measured outcomes variables. CONCLUSIONS: An assessment of preoperative pupil size and the attempted level of both SE and astigmatic correction may be useful in identifying patients at risk of developing symptoms or declines in visual performance after PARK. However, follow-up studies are indicated to identify variables predictive of poor visual outcomes following excimer laser refractive surgery. PMID- 11408141 TI - Diffuse lamellar keratitis: isolation of endotoxin and demonstration of the inflammatory potential in a rabbit laser in situ keratomileusis model. AB - PURPOSE: To systematically examine sources of endotoxin contamination in eye centers as a potential cause of diffuse lamellar keratitis (DLK) and to demonstrate the inflammatory potential of endotoxin in a rabbit model of laser in situ keratomileusis (LASIK) surgery. SETTING: University of Calgary, Calgary, Alberta, Canada. METHODS: In this prospective study, all water sources that routinely come in contact with LASIK instruments, including sterilizer reservoirs, eyedrops, microkeratome blades, and cleaning solutions, were examined for endotoxins at 5 eye centers. Bacterial cultures were performed on water samples from 5 sterilizer reservoirs. A LASIK flap was created in 8 rabbit eyes using an Automated Corneal Shaper microkeratome (Bausch & Lomb). The flaps were reflected, and a dose of endotoxin at various concentrations was placed on the interface. After 1 minute, the flap was irrigated and repositioned. The rabbit eyes were examined daily with a slitlamp biomicroscope for 3 days for the development of DLK, which was classified on a scale from grade 1 to 4 (mild to severe). The rabbits were killed at the conclusion of the study, and the interfaces were stained to rule out infectious etiologies. RESULTS: Endotoxin was detected in significant concentrations in tap water, filtered and distilled water, instrument washbasins, and sterilizer reservoirs at all 5 centers. The cultures of the water samples taken from the sterilizer reservoirs ranged from no growth to the presence of >100 colony-forming units of Flavobacterium and Pseudomonas aeruginosa. Endotoxins caused DLK-like interface inflammation in all eyes tested. Examination of stained scrapings showed no microorganisms in the interface of the rabbit eyes. CONCLUSION: Endotoxin contamination was detected in water sources that routinely come in contact with LASIK instruments. Endotoxins were capable of inducing interface inflammation in a rabbit model and may therefore be a significant factor in epidemic DLK. PMID- 11408142 TI - Direct measurement of microkeratome gap width by electron microscopy. AB - PURPOSE: To perform an accurate direct measurement of the microkeratome gap width using scanning electron microscopy (SEM). SETTING: Electron Microscope Unit, University of Hong Kong, Hong Kong, China. METHODS: The Cambridge Stereoscan S440 scanning electron microscope was used to measure the gap width of 4 SCMD microkeratomes with high accuracy (+/-1.5 microm). RESULTS: The manufacturer's gap specification for the 4 microkeratomes was 150.0 microm. The gap width measurements using SEM were 164.7 microm, 190.0 microm, 200.6 microm, and 145.9 microm and the respective errors, 9.8%, 26.7%, 33.7%, and 2.7%. Two of the 4 microkeratomes had more than a 25% error in gap width from the specification. CONCLUSIONS: The great variation in gap width from the manufacturer's specification for the 4 SCMD microkeratomes was beyond the standard of tolerance normally accepted in laser in situ keratomileusis (LASIK). Many unexpected LASIK related keratectasia and corneal perforations may be related to substandard microkeratome manufacturing and calibration. All new microkeratomes and blades should be validated before use to avoid keratectasia and other flap problems in LASIK. PMID- 11408143 TI - Intraocular lens designed for the newborn infant eye. AB - PURPOSE: To evaluate the effects of an intraocular lens (IOL) designed for small eyes after clear lens extraction in an animal model. SETTING: St. Erik's Eye Hospital, Karolinska Institute, Stockholm, Sweden. METHODS: Clear lens extraction was performed in both eyes in 19 3-week-old rabbits. In 1 randomly selected eye of each rabbit, a small IOL with long haptics was implanted. Axial length, corneal thickness, corneal diameter, and intraocular pressure were measured preoperatively and every month for 6 months postoperatively. Six months after surgery, the wet mass of the after-cataract was determined. RESULTS: The IOL remained well centered in all eyes. Four animals developed severe glaucoma in 5 eyes (3 aphakic and 2 pseudophakic) soon after surgery and were excluded. In the 15 animals completing the study, signs of glaucoma evolved in 7 animals (5 aphakic and 4 pseudophakic eyes). In aphakic eyes, significant amounts of after cataract (median 250 mg) developed in all 15 surviving animals. In pseudophakic eyes, small amounts of after-cataract (median 30 mg) were present. During the first 2 months after surgery, ocular growth was less in pseudophakic eyes than in aphakic eyes. CONCLUSIONS: Results indicate that implantation of a down-sized IOL with long haptics in small eyes is safe. Considerable less after-cataract was found in eyes with the IOL than in aphakic control eyes. PMID- 11408144 TI - Feasibility of ultrasound cataract surgery with a 1.4 mm incision. AB - PURPOSE: To study the feasibility of performing ultrasound (US) phacoemulsification cataract surgery through a 1.4 mm incision using conventional phacoemulsification equipment but removing the infusion sleeve from the US tip. SETTING: Department of Ophthalmology, Jikei University, Tokyo, Japan. METHODS: The infusion sleeve was removed from a 20 gauge US tip, and the sleeveless tip was inserted in a 1.4 mm incision in a postmortem porcine eye, providing infusion through a side port; phacoemulsification was performed with the US tip occluded. Temperature at the incision site was measured with a thermometer to determine whether a thermal burn occurred during the process. A hooked infusion cannula with a widened inner channel and 3 apertures was used to stabilize the anterior chamber depth. RESULTS: Ultrasound phacoemulsification produced almost no temperature elevation at the incision site as long as the infusion liquid was adequately circulated around the US tip. With the 20 gauge US tip, an adequate volume of leakage was maintained through the 1.4 mm incision; no thermal burns developed at the incision site. The use of a hooked infusion cannula made it possible to stabilize the anterior chamber and to apply the bimanual nucleofractis technique to emulsify and aspirate the lens nucleus. CONCLUSION: Using a sleeveless 20 gauge US tip, US cataract surgery was safely performed through a 1.4 mm incision without producing thermal burns at the incision site. PMID- 11408145 TI - Corneal endothelial cytotoxicity of diluted povidone--iodine. AB - PURPOSE: To assess corneal endothelial toxicity of diluted povidone-iodine (PI) in vivo and in vitro. SETTING: Cell Biology Laboratory and the Laboratory for Intraocular Microsurgery and Implants, Goldschleger Eye Research Institute, Sackler School of Medicine, Tel-Aviv University, Chaim Sheba Medical Center, Tel Hashomer, Israel. METHODS: In an in vitro study, cultured bovine corneal endothelial cells were exposed to diluted PI. The degree of cell damage was determined by staining with trypan blue and by comparing the results to those in a control group. In an in vivo study, a single dose of diluted PI was injected into the anterior chamber of rabbit eyes, completely replacing the aqueous humor. The eyes were evaluated by clinical examination, specular microscopy, pachymetry, pneumotonometry, and histopathology and compared to a control group injected with a balanced salt solution. RESULTS: In vitro, PI concentrations of 0.05% or less did not induce endothelial cell damage. Significant damage was observed with a PI concentration of 0.1%. Calf serum concentrations of 1% and higher in the culture media protected the endothelial cell monolayer from cytotoxic damage by PI. Aqueous humor did not have a similar effect. In vivo, PI concentrations of 0.1% or less did not induce changes in corneal endothelium morphology or function as assessed by specular microscopy and pachymetry. A PI concentration of 1% served as a positive control, causing corneal edema and endothelial cell loss as demonstrated by pachymetry, histopathology, and elevated intraocular pressure. CONCLUSIONS: The concentrations of PI tolerated by animal endothelium in vitro and in vivo were higher than the reported bactericidal levels. These findings justify further investigation of the safety and efficacy of PI for intracameral prophylaxis during surgery. PMID- 11408146 TI - Practice styles and preferences of ASCRS members--2000 survey. American Society of Cataract and Refractive Surgery. AB - A survey of the practice styles and preferences of members of the American Society of Cataract and Refractive Surgery with a United States ZIP code was performed in August 2000. Approximately 26% (1400) of 5342 questionnaires mailed were returned prior to the november cutoff date. Three profile questions were used to cross-tabulate: age of the ophthalmologist, geographic location, and volume of cataract surgery per month. Current data were compared with data in previous surveys. PMID- 11408147 TI - Ciliary body tumor and cataract: local resection combined with phacoemulsification. AB - A 57-year-old man developed a ciliary body mass, clinically diagnosed as malignant melanoma of the ciliary body, that produced a cataract in the right eye. Treatment was cataract surgery with sclerouvectomy performed simultaneously. Pathohistologic examination revealed an acquired adenoma of the nonpigmented ciliary epithelium. The clinicopathologic features and treatment of this tumor are discussed. PMID- 11408148 TI - Respiratory arrest and convulsions after peribulbar anesthesia. AB - We present a case in which a rare and serious complication with respiratory arrest and convulsions occurred after peribulbar anesthesia for cataract surgery. To our knowledge, respiratory arrest has been described as a complication only of retrobulbar, not peribulbar, anesthesia. Anesthesiologists trained in airway maintenance and ventilatory support should be available for immediate help when a peribulbar block is performed. PMID- 11408150 TI - Human hepatocyte culture systems for the in vitro evaluation of cytochrome P450 expression and regulation. AB - Primary cultures of human hepatocytes have been used extensively by both academic and industrial laboratories for evaluating the hepatic disposition of drugs and other xenobiotics. Their primary utility has been for assessing the induction potential of new chemical entities (NCEs) and they continue to serve as the gold standard. Primary considerations for conducting in vitro drug testing utilizing cultures of human hepatocytes, such as the effects of culture and study conditions, are discussed. The maintenance of normal cellular physiology and intercellular contacts in vitro is of particular importance for optimal phenotypic gene expression and response to drugs and other xenobiotics. Significant advances in our understanding of cytochrome P450 (CYP450) enzyme regulation have been made with the recent identification of the nuclear receptors mediating the induction of CYP2B and CYP3A enzymes. In particular, the activation of pregnane X receptor (PXR) by prototypical inducers of CYP3A has been found to correlate well with the species-specific modulation of CYP3A by various drugs and other xenobiotics. Concomitant with the discovery of PXR has been the identification of compounds that may act synergistically or antagonistically on multiple receptors (e.g., co-repressors and/or co-activators of the receptor) introducing novel mechanisms of drug-drug interactions. Differential expression of the individual isoforms of the major CYP450 enzymes over time in culture suggest that this model system is not reflective of in vivo profiles and, therefore, may be limited in its application for drug metabolism studies. Overall, primary cultures of human hepatocytes can serve as a sensitive and selective model for predicting the regulation of CYP450 modulation by drugs and other xenobiotics. Considerations and recommendations for standardizing testing conditions and choosing relevant endpoint(s) are presented. PMID- 11408151 TI - A scintigraphic study to investigate the potential for altered gut distribution of loperamide from a loperamide-simethicone formulation in man. AB - A loperamide simethicone combination formulation has recently been demonstrated to have significant clinical advantages compared to loperamide alone in the relief of diarrhoea and related symptoms. The product visualisation technique of gamma scintigraphy has been used to investigate the interaction of the formulation with the heterogenous environment of the human gut in a group of 12 healthy volunteers. The results suggest that changes in the intestinal kinetics of loperamide from the combination product, e.g. jejunal coating, could be contributing to the improved efficacy. PMID- 11408152 TI - Evaluation of the Intelisite capsule to deliver theophylline and frusemide tablets to the small intestine and colon. AB - The objective of the research was to establish the capability of the Intelisite capsule to deliver the probe drugs, theophylline and frusemide, in the form of split immediate release (IR) tablets, to the small intestine and colon. The two probe drugs were administered together in an open, random, three-way crossover study in eight healthy volunteers, comparing absorption following Intelisite delivery in the small bowel and colon to conventional IR dosing. Gamma scintigraphy was employed to monitor the gastrointestinal transit and activation of the Intelisite capsule. Standard pharmacokinetic parameters, and the percentage remaining in the capsules post defecation were determined. The Intelisite capsule was well tolerated in human volunteers and successfully activated on 15/16 occasions. Pharmacoscintigraphy showed internal marker release from the Intelisite capsule to be approximately 10-fold faster in the small intestine than in the colon. Theophylline and frusemide were both well absorbed following Intelisite activation in the small intestine, whereas complete colonic absorption was only observed in 1/7 subjects for theophylline, and 0/7 subjects for frusemide. The probe drugs were successfully delivered in particulate form from the Intelisite capsule in the small intestine and produced expected pharmacokinetic profiles. However drug release in the colon was incomplete and variable possibly due to: low water content, poor mixing, and a high loading dose. PMID- 11408153 TI - A flow-through optosensing device with fluorimetric transduction for rapid and sensitive determination of dipyridamole in pharmaceuticals and human plasma. AB - A flow-through optosensor with fluorimetric transduction has been prepared for the sensitive and selective determination of dipyridamole in aqueous solutions and biological fluids. The method is based on a monochannel flow-injection analysis system using Sephadex QAE A-25 resin, placed into a Hellma 176-QS fluorimetric flow-through cell, as an active sorbing substrate. The native fluorescence of dipyridamole fixed on the solid sorbent is continuously monitored at wavelengths of 305 and 490 nm for excitation and emission, respectively. After obtaining the maximum fluorescence intensity, the eluent solution (KH(2)PO(4)/NaOH buffer solution, c(T)=0.05 mol l(-1), pH 6.0) is allowed to reach the flow cell, the analyte is removed, and the resin support is regenerated. When an NaOH (10(-4) mol l(-1))/NaCl (0.1 mol l(-1)) solution is used as carrier solution, at a flow-rate of 1.56 ml min(-1), the sensor responds linearly in the measuring range of 10-500 microg l(-1) with a detection limit of 0.94 microg l(-1) and a throughput of 22 samples per hour (300 microl of sample volume). The relative standard deviation for ten independent determinations (200 microg l(-1)) is less than 0.82%. The method was satisfactorily applied to the determination of dipyridamole in pharmaceutical preparations and human plasma. PMID- 11408154 TI - Physicochemical characterisation of a drug-containing phospholipid-stabilised o/w emulsion for intravenous administration. AB - Clomethiazole (CMZ) was used as a model drug to be incorporated into an emulsion vehicle. The effects of drug concentration and number of homogenisation steps were evaluated using multiple linear regression. The droplet size, measured as a z-average diameter by photon correlation spectroscopy (PCS), was found to be between 60 and 260 nm in the investigated range of CMZ concentrations, highly dependent on the concentration, but more weakly so on the number of homogenisation steps. Slow-scanning high-sensitivity differential scanning calorimetry (DSC) measurements showed that CMZ depresses the phospholipid chain melting temperature in the emulsion system, whereas (13)C nuclear magnetic resonance (NMR) experiments suggested that the CMZ molecules are to a large extent located in the surface region of the emulsion droplets. This interpretation is compatible with results from NMR self-diffusion measurements, which showed that most of the CMZ molecules are rapidly exchanged between emulsion droplets and the aqueous surrounding. It can be concluded that the surface-active drug CMZ has a significant influence on the characteristics of phospholipid-stabilised emulsions through its ability to interact with the phospholipid interface. Thus, the results underline the importance of characterising drug-lipid interactions for the development of lipid-based formulations. PMID- 11408155 TI - Addition of hydrogen bond donating excipients to oil solution: effect on in vitro drug release rate and viscosity. AB - In oily vehicles containing different hydrogen bond donating excipients rates of transfer of the weak electrolytes naproxen and lidocaine from the oil phase to the aqueous medium were measured by using the rotating dialysis cell. A logarithmic linear correlation was established between the apparent partition coefficient, P(app), and the first-order rate constant related to attainment of equilibrium between the two phases, k(obs), which fitted well with results from former publications. Further, release data for the non-electrolyte testosterone were found to fit into this linear correlation. Apparent partition coefficients were determined between oil vehicles containing various amounts of hydrogen bond donating excipients and phosphate buffer, pH 6.00, revealing a rise in log P(app) with increasing concentration of excipient. Viscosity was measured for castor oil containing vehicles showing a linear relationship between percentage (v/v) castor oil and log viscosity (mPas) of the mixed vehicle. PMID- 11408156 TI - Magnetic marker monitoring of disintegrating capsules. AB - Magnetic marker monitoring was studied for its applicability to investigate the in vivo fate and behavior of disintegrating magnetically marked dosage forms. As a model, hard gelatin capsules were filled with an effervescent mixture of lactose, ascorbic acid and sodium hydrogen carbonate containing 1.3 mg black iron oxide as a magnetic label. The accuracy of the localization procedure whilst calculating all parameters of the dipole in one fitting procedure was checked in phantom experiments where the capsules were moved in well-defined paths with respect to the measurement device. The calculated position coordinates of the capsules deviated between less than 2 mm up to 8 mm from the expected position values depending on the distance between the sensor area and the capsule's path. Further experiments on the in vitro disintegration of the capsules showed that the value of the magnetic moment of the capsules can serve as a measure for their disintegration behavior. In vivo monitoring of the capsules was performed in eight experiments where a healthy volunteer swallowed each time one of the capsules. It was found that the in vivo disintegration behavior of the capsules corresponds well to their disintegration observed in water of about 37 degrees C. PMID- 11408157 TI - Investigation of the biological mode of action of clerocidin using whole cell assays. AB - Clerocidin, a diterpenoid natural product, has been shown in vitro to inhibit DNA religation following cleavage by topoisomerase II. Herein, we characterize the efficacy and specificity of clerocidin in HeLa cells. Our results suggest that clerocidin recognizes topoisomerase II as its main intracellular target and binds to this enzyme prior to formation of the 'cleavable complex' with DNA. These pharmacological features attest to the promising chemotherapeutic potential of this natural product. PMID- 11408158 TI - Signaling pathways and effector mechanisms pre-programmed cell death. AB - Apoptosis is a complex biochemical process that involves all aspects of the cell from the plasma membrane to the nucleus. Apoptosis stimuli are mediated by many different cellular processes including protein synthesis and degradation, the alteration in protein phosphorylation states, the activation of lipid second messenger systems, and disruption of normal mitochondrial function. Despite this diversity in signal transduction, all apoptotic pathways are believed to converge ultimately with the activation of caspases leading to the characteristic morphological changes of apoptosis. In this review, we discuss what is known about these pathways and its implication for normal cellular function. PMID- 11408159 TI - Effect of C-ring modifications in benzo[c]quinolizin-3-ones, new selective inhibitors of human 5 alpha-reductase 1. AB - The synthesis and the inhibition potency of octahydro- and decahydrobenzo[c]quinolizin-3-one derivatives 3--7, as new non-steroidal selective inhibitors of human enzyme 5 alpha-reductase type 1, are reported. These compounds differ from the recently reported benzo[c]quinolizin-3-one inhibitors 2 by the presence of a fully or partially saturated C-ring. Compounds 3 and 4, with a double bond in the C-ring, were prepared by sequential rearrangement-annulation of isoxazolines 19 and 20. C-ring saturated compounds 5- 7 were prepared by the Lewis acid-promoted Mannich-Michael tandem reaction of Danishefsky diene with the appropriate N-t-Boc iminium ion. Inhibition experiments were carried out on 5 alpha R-1 and 5 alpha R-2 expressed by CHO cells. Among the prepared compounds, octahydrobenzo[c]quinolizin-3-one 3, with a double bond at the position 6a--10a, was a potent and selective inhibitor of human 5 alpha R-1 (IC(50)=58 nM). The introduction of a tert-butylcarboxyamide at the position 8 (compound 4) was deleterious for the inhibition activity. The lack of the double bond in the C-ring reduced strongly the inhibition activity of compounds 5--7. The extended planarity of the most potent benzo[c]quinolizin-3 ones as well as favorable interactions of the C-ring unsaturation with the enzyme active site could account for the inhibition activity of these compounds. PMID- 11408160 TI - Novel mimics of sialyl Lewis X: design, synthesis and biological activity of a series of 2- and 3-malonate substituted galactoconjugates. AB - A series of potent inhibitors of P-selectin as potential anti-inflammatory agents is reported. These compounds are derivatives of galactocerebrosides bearing a malonate side chain in positions 2 and 3 of the galactose moiety. Based on the binding mode of sialyl Lewis X, the two acidic groups of the malonate are designed to form ionic interactions with two important lysines in the active site of P-selectin, Lys113 and Lys111. On the other hand, the 4- and 6-hydroxy groups on the galactose ring are arranged to chelate the calcium ion in the P-selectin active site. The synthesis and the biological activity of this series of compounds are described. Lead compounds having a greater potency than sialyl Lewis X are identified. PMID- 11408161 TI - Dicaffeoyl- or digalloyl pyrrolidine and furan derivatives as HIV integrase inhibitors. AB - Human immunodeficiency virus (HIV) integrase (IN) catalyzes the integration of HIV DNA copy into the host cell DNA. Such integration is essential for the production of progeny viruses, and therefore therapeutic agents that can inhibit this process should be effective anti-HIV agents. We have previously reported the inhibitory activity of dicaffeoylglucosides against HIV IN. In the present study, we have synthesized and tested dicaffeoyl or digalloyl compounds joined through a five-membered heterocyclic ring as HIV IN inhibitors to explore the SARs of this family of compounds. The starting heterocyclic diols were prepared from L tartaric acid, diethyl L-tartarate or D-(+)-ribonic gamma-lactone. We found that the HIV IN inhibitory activities of dicaffeoyl derivatives were comparable to that of L-chicoric acid (IC(50)=24.9 microM). On the other hand, digalloyl derivatives were more potent than L-chicoric acid with IC(50) values of 4.7--15.6 microM. PMID- 11408162 TI - N-terminal carboxyl and tetrazole-containing amides as adjuvants to Grb2 SH2 domain ligand binding. AB - High affinity binding of peptides to Src homology 2 (SH2) domains, often requires the presence of phosphotyrosyl (pTyr) or pTyr-mimicking moieties in the N terminal position of the binding ligand. Several reports have shown that N(alpha) acylation of the critical pTyr residue can result in increased SH2 domain binding potency. For Grb2 SH2 domains which recognize pTyr-Xxx-Asn-NH(2) motifs, significant potency enhancement can be incurred by N(alpha)-(3-amino)Z derivatization of tripeptides such as pTyr-Ile-Asn-NH(2). Using ligands based on the high affinity pY-Ac(6)c-Asn-(naphthylpropylamide) motif, (where Ac(6)c=1 aminocyclohexanecarboxylic acid), additional reports have shown moderate potentiating effects of N(alpha)-oxalyl derivatization. The current study examined variations of the N(alpha)-oxalyl theme in the context of a Xxx-Ac(6)c Asn-(naphthylpropylamide) platform, where Xxx=the hydrolytically stable pTyr mimetics phosphonomethyl phenylalanine (Pmp) or carboxymethyl phenylalanine (Cmf). The effects of N(alpha)-(3-amino)Z derivatization were also investigated for this platform, to ascertain whether the large binding enhancement reported for tripeptides such as pTyr-Ile-Asn-NH(2) could be observed. In ELISA-based extracellular Grb2 SH2 domain binding assays, it was found for the Pmp-based series, that extending the oxalyl carboxyl out by one methylene unit or replacing carboxyl functionality with a tetrazole isostere, resulted in binding potency greater than the parent N(alpha)-acetyl-containing compound, with enhancement approximating that observed for the N(alpha)-oxalyl derivative. When Cmf was used as the pTyr mimetic, only modest differences in IC(50) values were observed for the series. Examination of the N(alpha)-(3-amino)Z derivatized Pmp-Ac(6)c-Asn (naphthylpropylamide), showed that binding affinity was reduced relative to the parent N(alpha)-acetyl analogue, in contrast to the reported significant enhancement of affinity observed with other peptide ligands. Treatment of MDA-453 tumor cells, which are mitogenically driven through erbB-2 tyrosine kinase dependent pathways, with Pmp-containing inhibitors resulted in growth inhibition, with the N(alpha)-oxalyl and N(alpha)-malonyl-containing compounds exhibiting IC(50) values (4.3 and 4.6 microM, respectively) approximately five-fold lower than the parent N(alpha)-acetyl-containing compound. Tetrazole and N(alpha)-(3 amino)Z-containing inhibitors were from two- to four-fold less potent than these latter analogues in the growth inhibition assays. PMID- 11408163 TI - Synthesis and preliminary pharmacological evaluation of 5-hydroxy- and 5,6 dihydroxy-1,2,3,7,12,12a-hexahydrobenzo[5,6]cyclohepta[1,2,3-ij]isoquinoline derivatives as dopamine receptor ligands. AB - A series of 5-hydroxy- and 5,6-dihydroxy-1,2,3,7,12,12a hexahydrobenzo[5,6]cyclohepta[1,2,3-ij]isoquinoline derivatives (5a--e and 6a--e) were synthesized as conformationally rigid analogues of 1 benzyltetrahydroisoquinoline and evaluated for their affinity at D(1) and D(2) dopamine receptors. All compounds showed lower D(1) and D(2) affinities than dopamine. The 5-hydroxy-1-methyl-2,3,12,12a-hexahydrobenzo[5,6]cyclohepta[1,2,3 ij]isoquinoline 5a and the 5,6-dihydroxy analogue 6a showed D(2) agonist activity. This was proved by their effects on prolactin release from primary cultures of rat anterior pituitary cells. Molecular modeling studies showed that the geometric parameters (namely the distances from meta and para hydroxyl oxygens to the nitrogen and the height of nitrogen from the hydroxylated phenyl ring plane) of the dopaminergic pharmacophore embedded in our compounds have lower values in comparison with those observed in D(1) and D(2) selective ligands. PMID- 11408164 TI - Selenotyrosine and related phenylalanine derivatives. AB - A new series of Se-substituted phenylalanine derivatives has been synthesized having the para position of the phenyl ring substituted by selenocyanate (-SeCN), seleninic acid (-SeO(2)H), or selenol (-SeH) functional groups. The starting material for synthesis was 4'-aminophenylalanine, which is readily available in DL- or L- forms. Selenium was incorporated into the ring by reacting the unprotected amino acid with nitrous acid, followed by reaction of the diazotized aromatic amine with potassium selenocyanate at pH 4-5 to give phenylalanine selenocyanate. The selenocyanate derivative was converted to the selenol directly by reduction with sodium borohydride, or oxidized to the seleninic acid, which was then reduced to the selenol. Alkylation of the selenol ('selenotyrosine') gave the selenoether derivatives of phenylalanine [(Phe-SeR), R=methyl or allyl], and air oxidation of the selenol gave the diselenide. Mild oxidation of the selenoether 4'-(MeSe)Phe with peroxide gave the selenoxide derivative, 4' [Se(O)Me]. Because of their stability and useful redox properties, aromatic selenoamino acids can be used as synthetic analogues to increase chemical functionality in proteins or peptides, and have potential pharmaceutical or nutritional applications. The possibility that aromatic selenoamino acids could be formed metabolically through reactions of reactive selenium intermediates with aromatic amino acid residues is discussed. PMID- 11408165 TI - Molecular cloning, expression and characterization of the first three genes in the mevalonate-independent isoprenoid pathway in Streptomyces coelicolor. AB - The mevalonate-independent biosynthetic pathway to isopentenyl diphosphate and dimethylallyl diphosphate, the universal precursors to the isoprenoids, operates in eubacteria, including Escherichia coli, in algae, and in the plastids of higher plants. A search of the Sanger Centre Streptomyces coelicolor genome database revealed open reading frames with ca. 40--50% identity at the deduced amino acid level to the first three E. coli enzymes of this pathway, corresponding to deoxyxylulose phosphate synthase, deoxyxylulose phosphate reductoisomerase and 2-C-methyl erythritol 4-phosphate cytidylyltransferase. The S. coelicolor genes have been cloned and expressed in E. coli, and the recombinant proteins characterized physically and kinetically. The presence of the corresponding enzyme activities in extracts of S. coelicolor CH999 further supports the operation of the mevalonate-independent pathway in this organism. PMID- 11408166 TI - Studies on selectin blocker. 9. SARs of non-sugar selectin blocker against E-, P , L-selectin bindings. AB - As a part of study of selectin blockers, we have already reported that a non sugar selectin antagonist (3) was successfully discovered using a computational screening (Hiramatsu, Y.; Tsukida, T.; Nakai, Y.; Inoue, Y.; Kondo, H. J. Med. Chem. 2000, 43, 1476). To investigate the SARs of compound 3 against E-, P-, and L-selectins, we synthesized the derivatives of compound 3 and evaluated their inhibitory activities toward selectin bindings. The structural diversity of compound 3 contained the following: (1) a modification of the spacer unit (4--7), (2) a modification of the tail unit (8--11), (3) a modification of the head unit (12--18). As a result, it was found that a non-sugar based selectin blocker (3) could be a potential lead compound for E-, P-, and L-selectin blockers and some of the derivatives showed broad and/or selective inhibitory activities toward the E-, P-, and L-selectins. In addition, it was found that the experimental evidence well supported that the computational screening using 3D-pharmacophore model could be useful methodology to find out a new lead for the several type of selectin blockers, which included a broad and/or a selective inhibitor. PMID- 11408167 TI - Synthesis and DNA nicking studies of a novel cyclic peptide: cyclo[Lys-Trp-Lys Ahx-]. AB - Two novel cyclic tetrapeptides: cyclo[Lys-Tyr-Lys-Ahx-] 7a and cyclo[Lys-Trp-Lys Ahx-] 7b were synthesized by coupling protected amino acid in solution and the subsequent cyclization effected by the pentafluorophenyl ester method as described in previous papers. These cyclic peptides were designed and synthesized to study their interaction with DNA, based on previous reports that linear peptides Lys-Tyr-Lys and Lys-Trp-Lys could bind to various forms of DNA and cleaved supercoiled DNA at apurinic sites. Ethidium bromide displacement assay showed that the apparent DNA binding constant of linear Lys-Tyr-Lys and cyclic peptide 7a are far below 1 x 10(3) M(-1), whereas those of cyclic peptide 7b and linear Lys-Trp-Lys are 1.9 x 10(4) M(-1) and 9.5 x 10(3) M(-1), respectively. Kinetic studies using agarose gel electrophoresis showed that cyclic peptide 7b and Lys-Trp-Lys possessed DNA nicking activity on natural supercoiled phi X174 DNA with nicking rate of 50.7 and 75.6 pM min(-1) at 65 degrees C, respectively, whereas cyclic peptide 7a and linear Lys-Tyr-Lys were devoid of the corresponding activity. The DNA nicking rate increased significantly with increase in reaction temperature. At reaction temperatures lower than 65 degrees C, the DNA nicking rate of cyclic peptide 7b exceeded that of linear Lys-Trp-Lys. The addition of 1 microM ferrous ion did not give significant enhancement effect on the DNA nicking rate by the peptides. UV irradiation gave a marked rate enhancement on the DNA nicking rate of linear Lys-Trp-Lys and a moderate enhancement on the DNA nicking rate of cyclic peptide 7b. PMID- 11408168 TI - Structures of withanosides I, II, III, IV, V, VI, and VII, new withanolide glycosides, from the roots of Indian Withania somnifera DUNAL. and inhibitory activity for tachyphylaxis to clonidine in isolated guinea-pig ileum. AB - Seven new withanolide glycosides called withanosides I, II, III, IV, V, VI, and VII were isolated from an Indian natural medicine, Ashwagandha, the roots of Indian Withania somnifera DUNAL. (Solanaceae), together with four known compounds, withaferin A, 5 alpha,20 alpha(F)(R)-dihydroxy-6 alpha,7 alpha-epoxy-1 oxowitha-2,24-dienolide, physagulin D, and coagulin Q. The structures of withanosides I, II, III, IV, V, VI, and VII were determined based on chemical and physicochemical evidence. Principal constituents, withanoside VI (10 and 30 microM) and withaferin A (10 microM), attenuated the tachyphylaxis to clonidine on electrically stimulated guinea-pig ileum in vitro. PMID- 11408169 TI - Synthesis and structure--mutagenicity relationship of benzo-annulated cyclopentaphenanthrenes. AB - The synthesis of 2,3-dihydro-1H-indeno[5,4-a]anthracene (2), the fluoreno[a]anthracenes 3 and 4, 2,3-dihydro-1H-cyclopenta[a]chrysene (6), 3,4 dihydro-2-vinylphenanthrene (10) and cyclopenta[c]chrysenes 11, 12 has been described. Structure analysis of the new products by (1)H and (13)C NMR spectroscopy is presented. Estimates of the mutagenic activity of compounds 2--4, 6 and 11--14 in Salmonella typhimurium determined by Ames' test indicate that all products are inactive for both TA 98 and TA 100 strains except 4,5-dihydro-3H cyclopenta[c]chrysene (12). The mutagenic properties of these compounds have been compared with those shown by previously studied benzo[g]cyclopenta[a]phenanthrenes and cyclopenta[c]phenanthrenes and discussed. Some conclusions have been drawn about the effects of benzoannulation and of the carbonyl function on the mutagenicity of this class of compounds. PMID- 11408170 TI - 3D-QSAR analysis of 2,4,5- and 2,3,4,5-substituted imidazoles as potent and nontoxic modulators of P-glycoprotein mediated MDR. AB - 3D-Quantitative structure-activity relationships of 2,4,5- and 2,3,4,5 substituted imidazoles as a novel class of potent and nontoxic modulators of Pgp mediated MDR were investigated using CoMFA and COMSIA approaches. The best CoMFA model obtained from 46 imidazole analogues is a two-component model with the following statistics. R(2)(cv)=0.643, RMSE(cv)=0.360 for the cross-validation, and R(2)=0.767, RMSE=0.290 for the fitted. The best COMSIA model obtained is also a two-component model with the following statistics. R(2)(cv)=0.619, RMSE(cv)=0.372 for the cross-validation, and R(2)=0.765, RMSE=0.292 for the fitted. PMID- 11408171 TI - Synthesis, chemical and enzymatic reactivity, and toxicity of dithymidylyl-3',5' phosphorofluoridate and -phosphorothiofluoridate. AB - Dithymidylyl-3',5'-phosphorofluoridate and phosphorothiofluoridate were obtained by fluorinolysis of the P--Se bond in appropriate bisdimethoxytrityl selenomethyl esters. These compounds, which are hydrolytically unstable, are not inhibitors of snake venom, spleen phosphodiesterases and alkaline phosphatase. Neither compound is highly toxic. PMID- 11408172 TI - Synthesis, cytotoxicity, DNA interaction and topoisomerase II inhibition properties of tetrahydropyrrolo[3,4-a]carbazole-1,3-dione and tetrahydropyrido [3,2-b]pyrrolo[3,4-g]indole-1,3-dione derivatives. AB - Three tetrahydropyrrolo[3,4-a]carbazole-1,3-diones (6--8) and two tetrahydropyrido[3,2-b]pyrrolo[3,4-g]indole-1,3-diones (11--12) have been synthesized. Their interaction with DNA was probed by absorption and thermal melting studies. Compounds 8 and 12 both equipped with a hydroxyethyl-aminoethyl side-chain demonstrated higher affinities for poly(dA-dT)(2) than compounds 6, 7 and 11 bearing a dimethylaminoethyl side-chain. Circular and electric linear dichroism measurements showed that all five drugs behave as typical DNA intercalating agents. A plasmid cleavage assay was used to evaluate the capacity of the drugs to inhibit human topoisomerase II. Compounds 8 and 12 which bind strongly to DNA were found to stabilize DNA-topoisomerase II covalent complexes but their topoisomerase II inhibitory properties do not correlate with their cytotoxic potential. Compounds 6 and 7 are essentially inactive whereas compounds 8, 11 and 12 exhibit a high toxicity to P388 murine leukemia cells and provoke a marked accumulation in the G2/M phase of the cell cycle. These compounds form a new class of DNA-targeted antitumor agents. PMID- 11408173 TI - Inhibition of serine proteases by functionalized sulfonamides coupled to the 1,2,5-thiadiazolidin-3-one 1,1 dioxide scaffold. AB - A challenge associated with drug design is the development of selective inhibitors of proteases (serine or cysteine) that exhibit the same primary substrate specificity, that is, show a preference for the same P(1) residue. While these proteases have similar active sites, nevertheless there are subtle differences in their S and S' subsites which can be exploited. We describe herein for the first time the use of functionalized sulfonamides as a design and diversity element which, when coupled to the 1,2,5-thiadiazolidin-3-one 1,1 dioxide scaffold yields potent, time-dependent inhibitors of the serine proteases human leukocyte elastase (HLE), proteinase 3 (PR 3) and cathepsin G(Cat G). Our preliminary findings suggest that (a) appending to the 1,2,5-thiadiazolidin-3-one 1,1 dioxide scaffold recognition and diversity elements that interact with both the S and S' subsites of a target protease may result in optimal enzyme selectivity and potency and, (b) functionalized sulfonamides constitute a powerful design and diversity element with low intrinsic chemical reactivity and potentially wide applicability. PMID- 11408174 TI - Melanocyte-Directed enzyme prodrug therapy (MDEPT): development of second generation prodrugs for targeted treatment of malignant melanoma. AB - Evaluation of second generation prodrugs for MDEPT, by oximetry, has highlighted structural properties that are advantageous and disadvantageous for efficient oxidation using mushroom tyrosinase. In particular, a sterically undemanding prodrug bis-(2-chloroethyl)amino-4-hydroxyphenylaminomethanone 28 was synthesised and found to be oxidised by mushroom tyrosinase at a superior rate to tyrosine methyl ester, the carboxylic acid of which is the natural substrate for tyrosinase. The more sterically demanding phenyl mustard prodrugs 9 and 10 were oxidised by mushroom tyrosinase at a similar rate to tyrosine methyl ester. In contrast, tyramine chain elongation via heteroatom insertion was detrimental and the rate of mushroom tyrosinase oxidation of phenyl mustard prodrugs 21 and 22 decreased by 10 nanomol/min. PMID- 11408175 TI - Synthesis, molecular modeling and QSAR studies in chiral 2,3-disubstituted 1,2,3,4-tetrahydro-9H-pyrido(3,4-b)indoles as potential modulators of opioid antinociception. AB - In view of coexistence of opioid and cholecystokinin (CCK) in the brain areas concerned with pain processing, some semirigid racemic and chiral analogues of a potent CCK receptor antagonist (benzotript) have been synthesized and tested for their modulatory role on opioid antinociception, which may be mediated by CCK-B receptor. Some of these compounds, 3e, 3g, 3h, 4a, 4b and 4h, exhibited antinociceptive potentiation comparable to benzotript and proglumide. In order to identify the essential chemical structural features important for this potentiation, molecular modeling and quantitative structure activity relationship (QSAR) studies have been carried out in the S and R enantiomers of some of these semi-rigid compounds. The 3D-biophore models, common to all molecules of the training set have been derived. These models with superimposition (match value >0.25) depicted three biophoric sites one each for, pi/hydrophobic interactions, hydrogen bonding and ionic interactions among the phenyl/pyrrole ring, indole nitrogen, amidic oxygen, pyridyl nitrogen and lone pair of amidic oxygen. The total hydrophobicity and S absolute stereochemistry are found to positively contribute to potentiation of antinociception induced by morphine and the resulting quantitative pharmacophoric model with good correlation is found to well describe the observed activity. PMID- 11408176 TI - Dicationic dithiocarbamate carbapenems with anti-MRSA activity. AB - A new class of 1 beta-methylcarbapenems bearing a doubly quaternarized 1,4 diazabicyclooctane (DABCO) substituted dithiocarbamate moiety at the C-2 side chain was prepared, and the biological profiles of the compounds, including in vitro and in vivo anti-MRSA activity and DHP-I susceptibility, were evaluated to identify a carbapenem derivative that was superior to BO-3482 (1). As a result, we discovered a 1 beta-methyl-2-[4-(4-carbamoylmethyl-1,4 diazabicyclo[2,2,2]octanediium-1-yl)methyl-1,2,3,6 tetrahydropyridinylthiocarbonylthio]carbapenem, 14a showing greater than 2-fold better anti-MRSA activity in a mouse infection model and 3-fold better DHP-I susceptibility as compared with BO-3482 (1). PMID- 11408177 TI - Identification of new triarylethylene oxyalkanoic acid analogues as bone selective estrogen mimetics. AB - Previously, the estrogen receptor (ER) ligand 4-[1-(p-hydroxyphenyl)-2 phenylethyl]phenoxyacetic acid (5) was found to have differential bone loss suppressive effects in the ovariectomized (OVX) rat approaching those of selective ER modulators (SERMs) such as tamoxifen. In an effort to improve efficacy, analogues of this compound were prepared which incorporated features designed to reduce polarity/ionizability. Thus, the acetic acid side chain of 5 was replaced by n-butanoic acid and 1H-tetrazol-4-ylmethyl moieties, to give 8 and 10, respectively. Also, the phenolic hydroxyl of 5 was replaced, giving deoxy analogue 9. We also developed new methods for the synthesis of triarylethylene variants of 5 and 9, namely 4-([1-(p-hydroxyphenyl)-2-phenyl-1-butenyl]phenoxy)-n butanoic acid (6) and its des-hydroxy counterpart (7), because the former of these had in vitro antiestrogenic effects characteristic of known SERMs. In the OVX rat, 6 and 7 were as effective as 17 beta-estradiol in suppressing serum markers of bone resorption/turnover, namely osteocalcin and deoxypyridinoline, but had only 30% of the uterotrophic efficacy of 17 beta-estradiol. This study has thus identified two triarylethylene oxybutyric acids, 6 and 7, that have differential bone/uterus effects like those of known SERMs. PMID- 11408178 TI - Design, synthesis, and biological evaluation of anti-HIV double-drugs. conjugates of HIV protease inhibitors with a reverse transcriptase inhibitor through spontaneously cleavable linkers. AB - Based on the prodrug concept as well as the combination of two different classes of anti-HIV agents, we designed and synthesized a series of anti-HIV double-drugs consisting of HIV protease inhibitors conjugated with a nucleoside reverse transcriptase inhibitor in an effort to enhance the antiviral activity. For the conjugation, a series of linkers that conjoins the two different classes of inhibitors has been investigated. Double-drugs using a succinyl amino acid linker were shown to release the parent drugs via spontaneous imide formation at a faster rate compared to compounds using a glutaryl amino acid linker, as expected from the energetically favorable cyclization to the five-membered ring. Among the double-drugs, KNI-1039 (3b) with a glutarylglycine linker exhibited extremely potent anti-HIV activity compared with that of the individual components. Double drug 3b was relatively stable in culture medium, whereas it regenerated active species in cell homogenate. These results suggested that the synergistic enhancement of anti-HIV activities of 3b may be due to their ability to penetrate into the target cell and subsequent regeneration of two different classes of anti HIV agents in the cytoplasm. PMID- 11408179 TI - Synthesis and antimicrobial activity of the symmetric dimeric form of Temporin A based on 3-N,N-di(3-aminopropyl)amino propanoic acid as the branching unit. AB - Dimeric derivative of antimicrobial peptide amide Temporin A (TA) was synthesized by using a new branching unit 3-N,N-di(3-aminopropyl)amino propanoic acid (DAPPA), which allows building of the parallelly symmetric alpha-helical structures. Antimicrobial effect of the original peptide amide, its monomeric carboxy (TAc) and novel dimeric (TAd) analogues were tested against Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative). Both TA and TAd completely inhibited the growth of S. aureus at the concentrations of 5 and 10 microM, respectively, whereas TAc did not show any inhibitory activity. The activities of TAc, TA and TAd correlate directly with the net charges of the molecules, +1, +2 and +4, respectively. Interestingly, TAd displayed antibacterial effect against E. coli at a concentration of 10 microM, where as monomeric TA did not show any activity at concentration as high as 20 microM. The results indicate that the novel structural modification improves the antibacterial properties of Temporin A especially towards Gram-negative bacteria. PMID- 11408181 TI - Synthesis and properties of triple helix-forming oligodeoxyribonucleotides containing 7-chloro-7-deaza-2'-deoxyguanosine. AB - Multiple incorporations of 7-chloro-7-deaza-2'-deoxyguanosine in place of 2' deoxyguanosine have been performed into a triple helix-forming oligodeoxyribonucleotide involving a run of six contiguous guanines designed to bind in a parallel orientation relative to the purine strand of the DNA target. The ability of these modified oligodeoxyribonucleotides to form triple helices in a buffer containing monovalent cations was studied by UV--melting curves analysis, gel shift assay and restriction enzyme protection assay. In the presence of Na(+), the incorporation of two, three or five modified nucleosides in the third strand has improved the efficacy of formation of the triplex as compared to that formed with the unmodified oligonucleotide. The stabilities of the three modified triplexes were similar. The coupling of 6-chloro-2-methoxy-9 (omega-hexylamino)-acridine to the 5'-end of the oligonucleotides containing modified nucleosides led to an increase in triplex stability similar to that observed when the acridine was added to the 5'-end of the unmodified oligonucleotide. In the presence of K(+), only the oligodeoxyribonucleotides containing modified G retained the ability to form triple helices with the same efficiency. The incorporation of the modified nucleoside has two effects: (i) it decreases TFO self-association, and (ii) it slightly increases triplex stability. The enhanced ability of the modified oligonucleotides containing 7-chloro-7-deaza 2'-deoxyguanosine over the parent oligomer to form triple helices was confirmed by inhibition of restriction enzyme cleavage using a circular plasmid containing the target sequence. PMID- 11408180 TI - Synthesis and phosphodiesterase 5 inhibitory activity of novel phenyl ring modified sildenafil analogues. AB - New sildenafil analogues containing an ether ring fused into the phenyl moiety, 6a--d and 7a--d, were efficiently synthesized from the readily available starting materials, 1a--d and 2, in five steps. Ab initio calculations indicated that introduction of a cyclic ether to the phenyl group might enhance the co-planarity of the molecule. The torsional angles were calculated to be 2--3 degrees for the 5-membered cyclic ether derivatives, 6a, 6c, 7a, and 7c, and 12--16 degrees for the 6-membered ones, 6b, 6d, 7b, and 7d. On the other hand, sildenafil showed the least co-planarity with the torsional angle of 23 degrees compared with the target compounds, 6a--d and 7a--d. In the enzyme assay, however, the in vitro PDE 5 inhibitory activity was found out to be inversely related to the degree of co planarity. In other words, the least planar sildenafil showed the highest activity, and the most planar 5-membered cyclic ether derivatives were least active by 100--200-fold compared with sildenafil. Our study clearly demonstrated that the open chain 2'-alkoxy group of the phenyl ring, although less effective for inducing the co-planarity, seemed to act as a much better lipophilic requirement than the cyclic alkoxy moiety. PMID- 11408182 TI - Introduction of the new dipeptide isostere 7-endo-BtA as reverse turn inducer in a Bowman-Birk proteinase inhibitor: synthesis and conformational analysis. AB - Two dipeptide isosteres 7-exo-BTG (1) and 7-endo-BtA (2), belonging to the new class of gamma/delta-bicyclic amino acid BTAa, were inserted into an 11-residue peptide deriving from the Bowman Birk Inhibitor (BBI) class of serine protease inhibitors, and the conformational properties of these modified peptides have been studied by NMR and molecular modelling. The dipeptide isostere 7-endo-BtA [(1R,4S,5R,7R)-4-endo-methyl-6,8-dioxa-3-azabicyclo[3.2.1]octane-7-endo carboxylic acid] (2), derived from L-alanine and meso tartaric acid, gave rise to the modified BBI peptide 5 whose structure was very similar to that of the original peptide 3, suggesting a possible reverse turn inducing property for this dipeptide isostere. PMID- 11408183 TI - Salen complexes with bulky substituents as useful tools for biomimetic phenol oxidation research. AB - The catalytic properties of bulky water-soluble Co-, Cu-, Fe- and Mn-salen complexes in the oxidation of phenolic lignin model compounds have been studied in aqueous water--dioxane solutions (pH 3--10). Mn catalysts were found to oxidize coniferyl alcohol in a same reaction time as horseradish peroxidase (HRP) enzyme and Mn and Co catalysts showed different regioselectivity suggesting a different substrate to catalyst interaction in the oxidative coupling. When the oxidation of material more relevant to plant polyphenolics was studied, the results indicated that the complexes catalyze one- and two-electron oxidations depending on the bulk of the substrate. PMID- 11408184 TI - Synthesis and study of a cyclic angiotensin II antagonist analogue reveals the role of pi*--pi* interactions in the C-terminal aromatic residue for agonist activity and its structure resemblance with AT(1) non-peptide antagonists. AB - The novel amide linked Angiotensin II (ANG II) cyclic analogue cyclo(3, 5) [Sar(1)-Lys(3)-Glu(5)-Ile(8)] ANG II (18) has been designed, synthesized and bioassayed in anesthetized rabbits. The constrained cyclic analogue with a lactam amide bridge linking a Lys-Glu pair at positions 3 and 5 and possessing Ile at position 8, was synthesized by solution procedure using the maximum protection strategy. This analogue was found to be inhibitor of Angiotensin II. NMR spectroscopy coupled with computational analysis showed clustering between the side chains of the key aminoacids Tyr(4)-His(6)-Ile(8) similar to that observed with ANG II. The obtained data show that only pi*--pi* interactions observed in ANG II or its superagonist Sar(1) [ANG II] are missing. Therefore, it can be concluded that these interactions are essential for agonist activity. Conformational analysis comparisons between AT(1) antagonists losartan, eprosartan and irbesartan with C-terminal segment of cyclic compound 18 revealed structural similarities. PMID- 11408187 TI - Neurons go to great lengths for new therapy. PMID- 11408186 TI - Have the ultimate benefits of clinical trials been maligned beyond repair? PMID- 11408188 TI - The end for diabetic kidney disease? PMID- 11408192 TI - Chinese pharmaceutical companies: an emerging industry. PMID- 11408189 TI - Underwater treasures for cancer treatment. PMID- 11408194 TI - More microwave reactors required. PMID- 11408193 TI - Communication networks: key to success of new technologies. AB - The Discussion Forum provides a medium for airing your views on any issues related to the pharmaceutical industry and obtaining feedback and discussion on these views from others in the field. You can discuss issues that get you hot under the collar, practical problems at the bench, recently published literature, or just something bizarre or humorous that you wish to share. Publication of letters in this section is subject to editorial discretion and company promotional letters will be rejected immediately. Furthermore, the views provided are those of the authors and are not intended to represent the views of the companies they work for. Moreover, these views do not reflect those of Elsevier, Drug Discovery Today or its editorial team. Please submit all letters to Dr Rebecca Lawrence, News & Features Editor, Drug Discovery Today, e-mail: Rebecca.Lawrence@current-trends.com PMID- 11408195 TI - Are drug targets missed owing to lack of physical activity? - Reply. PMID- 11408196 TI - New hope for matrix metalloproteinase inhibitors in cancer therapy. PMID- 11408197 TI - Mass spectrometry for the 21st century. PMID- 11408198 TI - Virogenomics: a novel approach to antiviral drug discovery. AB - Target discovery in virology has been limited to the few open-reading frames encoded by viral genomes. However, several recent examples show that inhibiting host-cell proteins can prevent viral infection. The human genome sequence should, therefore, contain many more genes that are essential for viral propagation than viral genomes. A systematic approach to find these potential cellular antiviral targets is global host gene expression analysis using DNA microarrays. Several recent studies reveal both unique and common strategies by which viruses change the gene expression profile of the host cell. Moreover, work in progress shows that some of the host pathways discovered by expression profiling are important for viral replication. Thus, human genomics tools have the potential to deliver novel antiviral drugs. PMID- 11408199 TI - Transgenic knockouts as part of high-throughput, evidence-based target selection and validation strategies. AB - The worldwide genome sequencing projects are helping to define the size and complexity of the expressed genome and are thereby identifying an unprecedented number of genes of uncertain disease alignment and unknown function. It is widely recognized that, within the pharmaceutical industry, a significant commercial advantage will accrue to those companies that most effectively gather and integrate additional biological information into their therapeutic target selection and drug progression strategies. This article presents the rationale for including comparative phenotypic information obtained from transgenic gene knockouts as an integral part of any future therapeutic target selection strategy. PMID- 11408200 TI - Miniaturized HTS technologies - uHTS. AB - The transition from slow, manual, low-throughput screening to industrialized robotic ultra-high throughput screening (uHTS) in the past few years has made it possible to screen hundreds of thousands of chemical entities against a biological target in a short time-frame. The need to minimize the cost of screening has been addressed primarily by reducing the volume of sample to be screened. This, in turn, has resulted in the miniaturization of HTS technology as a whole. Miniaturization requires new technologies and strategies for compound handling, assay development, assay adaptation, liquid handling and automation in addition to refinement of the technologies used for detection systems and data management. This review summarizes current trends in the field of uHTS and illustrates the technological developments that are necessary to enable the routine application of miniaturized uHTS systems within an industrial environment. PMID- 11408201 TI - Monitor: molecules and profiles. AB - Monitor provides an insight into the latest developments in drug discovery through brief synopses of recent presentations and publications together with expert commentaries on the latest technologies. There are two sections: Molecules summarizes the chemistry and the pharmacological significance and biological relevance of new molecules reported in the literature and on the conference scene; Profiles offers commentary on promising lines of research, emerging molecular targets, novel technology, advances in synthetic and separation techniques and legislative issues. PMID- 11408202 TI - Novel antitumour molecules. PMID- 11408204 TI - Erratum. PMID- 11408203 TI - Bone-specific delivery of diclofenac. PMID- 11408205 TI - Search for new drugs for treatment of tuberculosis. PMID- 11408206 TI - Effects of renal function on pharmacokinetics of recombinant human granulocyte colony-stimulating factor in lung cancer patients. AB - Animal studies suggest that the kidney is involved in the elimination of recombinant human granulocyte colony-stimulating factor (rhG-CSF), which is used for patients with neutropenia during cancer chemotherapy. Since anticancer drugs induce nephrotoxicity, it is important to clarify the role of the kidney in the pharmacokinetics of rhG-CSF in cancer patients. Our study was designed to evaluate the relationship between the pharmacokinetics of rhG-CSF and renal function in lung cancer patients compared to the absolute neutrophil count (ANC). The pharmacokinetic studies were conducted with 25 lung cancer patients. Following chemotherapy using platinum-based compounds, a bolus 5 microg of rhG CSF/kg of body weight was intravenously injected from the first day of leukopenia or neutropenia. Pharmacokinetic parameters were estimated by fitting the concentration in serum-time data to a two-compartment model according to the population pharmacokinetics and the Bayesian method. Creatinine clearance (CL(CR)) was predicted by the Cockcroft-Gault formula. rhG-CSF clearance (CL(G CSF)) correlated significantly with the ANC (r = 0.613; P < 0.001) and CL(CR) (r = 0.632; P < 0.001). Multiple linear regression analysis showed that the combination of the ANC and CL(CR) accounted for 57.4% of the variation of CL(G CSF). In patients with an ANC of <1,000/microl, CL(CR) accounted for 72.9% of the variation of CL(G-CSF) (P < 0.001). Our findings suggest that renal function and neutrophil counts correlate with CL(G-CSF) and that the role of renal function in eliminating rhG-CSF is important in lung cancer patients with neutropenia. PMID- 11408207 TI - In vitro activities of linezolid, meropenem, and quinupristin-dalfopristin against group C and G streptococci, including vancomycin-tolerant isolates. AB - The in vitro activities of meropenem, linezolid, quinupristin-dalfopristin, vancomycin, and penicillin against 130 clinical isolates of group C and G streptococci, including vancomycin-tolerant isolates, were evaluated. Meropenem, linezolid, quinupristin-dalfopristin, vancomycin, and penicillin MICs at which 90% of the isolates were inhibited were 0.06, 2.0, 0.25, 0.5, and < or = 0.016 microg/ml, respectively. Meropenem, linezolid, quinupristin-dalfopristin, and penicillin were active against group C and G streptococci, including vancomycin resistant strains. PMID- 11408208 TI - Genetic organization of the chromosome region surrounding mecA in clinical staphylococcal strains: role of IS431-mediated mecI deletion in expression of resistance in mecA-carrying, low-level methicillin-resistant Staphylococcus haemolyticus. AB - We report on the structural diversity of mecA gene complexes carried by 38 methicillin-resistant Staphylococcus aureus and 91 methicillin-resistant coagulase-negative Staphylococcus strains of seven different species with a special reference to its correlation with phenotypic expression of methicillin resistance. The most prevalent and widely disseminated mec complex had the structure mecI-mecR1-mecA-IS431R (or IS431mec), designated the class A mecA gene complex. In contrast, in S. haemolyticus, mecA was bracketed by two copies of IS431, forming the structure IS431L-mecA-IS431R. Of the 38 S. haemolyticus strains, 5 had low-level methicillin resistance (MIC, 1 to 4 mg/liter) and characteristic heterogeneous methicillin resistance as judged by population analysis. In these five strains, IS431L was located to the left of an intact mecI gene, forming the structure IS431L-class A mecA-gene complex. In other S. haemolyticus strains, IS431L was associated with the deletion of mecI and mecR1, forming the structure IS431L-DeltamecR1-mecA-IS431mec, designated the class C mecA gene complex. Mutants with the class C mecA gene complex were obtained in vitro by selecting strain SH621, containing the IS431L-class A mecA gene complex with low concentrations of methicillin (1 and 3 mg/liter). The mutants had intermediate level of methicillin resistance (MIC, 16 to 64 mg/liter). The mecA gene transcription was shown to be derepressed in a representative mutant strain, SH621-37. Our study indicated that the mecI-encoded repressor function is responsible for the low-level methicillin resistance of some S. haemolyticus clinical strains and that the IS431-mediated mecI gene deletion causes the expression of methicillin resistance through the derepression of mecA gene transcription. PMID- 11408209 TI - Pseudomonas aeruginosa reveals high intrinsic resistance to penem antibiotics: penem resistance mechanisms and their interplay. AB - Pseudomonas aeruginosa exhibits high intrinsic resistance to penem antibiotics such as faropenem, ritipenem, AMA3176, sulopenem, Sch29482, and Sch34343. To investigate the mechanisms contributing to penem resistance, we used the laboratory strain PAO1 to construct a series of isogenic mutants with an impaired multidrug efflux system MexAB-OprM and/or impaired chromosomal AmpC beta lactamase. The outer membrane barrier of PAO1 was partially eliminated by inducing the expression of the plasmid-encoded Escherichia coli major porin OmpF. Susceptibility tests using the mutants and the OmpF expression plasmid showed that MexAB-OprM and the outer membrane barrier, but not AmpC beta-lactamase, are the main mechanisms involved in the high intrinsic penem resistance of PAO1. However, reducing the high intrinsic penem resistance of PAO1 to the same level as that of penem-susceptible gram-negative bacteria such as E. coli required the loss of either both MexAB-OprM and AmpC beta-lactamase or both MexAB-OprM and the outer membrane barrier. Competition experiments for penicillin-binding proteins (PBPs) revealed that the affinity of PBP 1b and PBP 2 for faropenem were about 1.8- and 1.5-fold lower, than the respective affinity for imipenem. Loss of the outer membrane barrier, MexAB, and AmpC beta-lactamase increased the susceptibility of PAO1 to almost all penems tested compared to the susceptibility of the AmpC-deficient PAO1 mutants to imipenem. Thus, it is suggested that the high intrinsic penem resistance of P. aeruginosa is generated from the interplay among the outer membrane barrier, the active efflux system, and AmpC beta lactamase but not from the lower affinity of PBPs for penems. PMID- 11408210 TI - Safety and bactericidal activity of rifalazil in patients with pulmonary tuberculosis. AB - Rifalazil, also known as KRM-1648 or benzoxazinorifamycin, is a new semisynthetic rifamycin with a long half-life of approximately 60 h. Rifalazil has potent bactericidal activity against Mycobacterium tuberculosis in vitro and in animal models of tuberculosis (TB). Prior studies in healthy volunteers showed that once weekly doses of 25 to 50 mg of rifalazil were well tolerated. In this randomized, open-label, active-controlled phase II clinical trial, 65 subjects with sputum smear-positive pulmonary TB received one of the following regimens for the first 2 weeks of therapy: 16 subjects received isoniazid (INH) (5 mg/kg of body weight) daily; 16 received INH (5 mg/kg) and rifampin (10 mg/kg) daily; 17 received INH (5 mg/kg) daily plus 10 mg of rifalazil once weekly; and 16 received INH (5 mg/kg) daily and 25 mg of rifalazil once weekly. All subjects were then put on 6 months of standard TB therapy. Pretreatment and day 15 sputum CFU of M. tuberculosis were measured to assess the bactericidal activity of each regimen. The number of drug-related adverse experiences was low and not significantly different among treatment arms. A transient decrease in absolute neutrophil count to less than 2,000 cells/mm(3) was detected in 10 to 20% of patients in the rifalazil- and rifampin-containing treatment arms without clinical consequences. Decreases in CFU counts were comparable among the four treatment arms; however, the CFU results were statistically inconclusive due to the variability in the control arms. Acquired drug resistance did not occur in any patient. Studies focused on determining a maximum tolerated dose will help elucidate the full anti TB effect of rifalazil. PMID- 11408211 TI - gyrA mutations associated with quinolone resistance in Bacteroides fragilis group strains. AB - Mutations in the gyrA gene contribute considerably to quinolone resistance in Escherichia coli. Mechanisms for quinolone resistance in anaerobic bacteria are less well studied. The Bacteroides fragilis group are the anaerobic organisms most frequently isolated from patients with bacteremia and intraabdominal infections. Forty-four clinafloxacin-resistant and-susceptible fecal and clinical isolates of the B. fragilis group (eight Bacteroides fragilis, three Bacteroides ovatus, five Bacteroides thetaiotaomicron, six Bacteroides uniformis, and 22 Bacteroides vulgatus) and six ATCC strains of the B. fragilis group were analyzed as follows: (i) determination of susceptibility to ciprofloxacin, levofloxacin, moxifloxacin, and clinafloxacin by the agar dilution method and (ii) sequencing of the gyrA quinolone resistance-determining region (QRDR) located between amino acid residues equivalent to Ala-67 through Gln-106 in E. coli. Amino acid substitutions were found at hotspots at positions 82 (n = 15) and 86 (n = 8). Strains with Ser82Leu substitutions (n = 13) were highly resistant to all quinolones tested. Mutations in other positions of gyrA were also frequently found in quinolone-resistant and -susceptible isolates. Eight clinical strains that lacked mutations in their QRDR were susceptible to at least two of the quinolones tested. Although newer quinolones have good antimicrobial activity against the B. fragilis group, quinolone resistance in B. fragilis strains can be readily selected in vivo. Mutational events in the QRDR of gyrA seem to contribute to quinolone resistance in Bacteroides species. PMID- 11408212 TI - Inducible macrolide resistance in Corynebacterium jeikeium. AB - Corynebacterium jeikeium is an opportunistic pathogen primarily of immunocompromised (neutropenic) patients. Broad-spectrum resistance to antimicrobial agents is a common feature of C. jeikeium clinical isolates. We studied the profiles of susceptibility of 20 clinical strains of C. jeikeium to a range of antimicrobial agents. The strains were separated into two groups depending on the susceptibility to erythromycin (ERY), with one group (17 strains) representing resistant organisms (MIC > 128 microg/ml) and the second group (3 strains) representing susceptible organisms (MIC < or = 0.25 microg/ml). The ERY resistance crossed to other members of the macrolide-lincosamide streptogramin B (MLSb) group. Furthermore, this resistance was inducible with MLSb agents but not non-MLSb agents. Expression of ERY resistance was linked to the presence of an allele of the class X erm genes, erm(X)cj, with >93% identity to other erm genes of this class. Our evidence indicates that erm(X)cj is integrated within the chromosome, which contrasts with previous reports for the plasmid-associated erm(X) genes found in C. diphtheriae and C. xerosis. In 40% of C. jeikeium strains, erm(X)cj is present within the transposon, Tn5432. However, in the remaining strains, the components of Tn5432 (i.e., the erm and transposase genes) have separated within the chromosome. The rearrangement of Tn5432 leads to the possibility that the other drug resistance genes have become included in a new composite transposon bound by the IS1249 elements. PMID- 11408213 TI - Durability of anti-infective effect of long-term silicone sheath catheters impregnated with antimicrobial agents. AB - This study was performed to test the long-term antimicrobial efficacy of impregnated silicone catheters comprising an antimicrobial layer sandwiched between an external surface sheath and a luminal surface silicone sheath. The design of the catheter permits the introduction of various antimicrobials in addition to anticoagulants or antifibrins in the antimicrobial layer and allows their gradual release over a period of months after insertion. The in vitro data presented show that the catheter can provide antimicrobial activity for 90 days, after being replated for 15 7-day cycles of replating. When the catheters were immersed in human serum and incubated at 37 degrees C, they demonstrated significant antimicrobial activity after more than 325 days of incubation. The significant long-term in vitro antimicrobial activity observed may imply effective in vivo activity for almost 1 year after insertion and could serve as a cost-effective alternative to surgically implantable silicone catheters. PMID- 11408214 TI - Interaction between DNA gyrase and quinolones: effects of alanine mutations at GyrA subunit residues Ser(83) and Asp(87). AB - DNA gyrase is a target of quinolone antibacterial agents, but the molecular details of the quinolone-gyrase interaction are not clear. Quinolone resistance mutations frequently occur at residues Ser(83) and Asp(87) of the gyrase A subunit, suggesting that these residues are involved in drug binding. Single and double alanine substitutions were created at these positions (Ala(83), Ala(87), and Ala(83) Ala(87)), and the mutant proteins were assessed for DNA supercoiling, DNA cleavage, and resistance to a number of quinolone drugs. The Ala(83) mutant was fully active in supercoiling, whereas the Ala(87) and the double mutant were 2.5- and 4- to 5-fold less active, respectively; this loss in activity may be partly due to an increased affinity of these mutant proteins for DNA. Supercoiling inhibition and cleavage assays revealed that the double mutant has a high level of resistance to certain quinolones while the mutants with single alanine substitutions show low-level resistance. Using a drug-binding assay we demonstrated that the double-mutant enzyme-DNA complex has a lower affinity for ciprofloxacin than the wild-type complex. Based on the pattern of resistance to a series of quinolones, an interaction between the C-8 group of the quinolone and the double-mutant gyrase in the region of residues 83 and 87 is proposed. PMID- 11408215 TI - MexXY-OprM efflux pump is required for antagonism of aminoglycosides by divalent cations in Pseudomonas aeruginosa. AB - Antagonism of aminoglycosides by divalent cations is well documented for Pseudomonas aeruginosa and is regarded as one of the problems in aminoglycoside therapy. It is generally considered that divalent cations interfere with uptake of aminoglycosides at both the outer and inner membranes. It has been demonstrated recently that aminoglycosides can be removed from cells of P. aeruginosa by the three-component multidrug resistance efflux pump MexXY-OprM. We sought to investigate the interplay between efflux and uptake in resistance to aminoglycosides in P. aeruginosa. To do so, we studied the effects of the divalent cations Mg(2+) and Ca(2+) on susceptibility to aminoglycosides in a wild type strain of P. aeruginosa and in mutants either overexpressing or lacking the MexXY-OprM efflux pump. MICs of gentamicin, streptomycin, amikacin, apramycin, netilmicin, and arbekacin were determined in Mueller-Hinton broth in the presence of cations added at concentrations that varied from 0.125 to 8 mM. We found, unexpectedly, that while both Mg(2+) and Ca(2+) antagonized aminoglycosides (up to a 64-fold decrease in susceptibility at 8 mM), antagonism was seen only in the strains of P. aeruginosa that contained the functional MexXY-OprM efflux pump. Our results indicate that inhibition of the MexXY-OprM efflux pump should abolish the antagonism of aminoglycosides by divalent cations, regardless of its precise mechanism. This may significantly increase the therapeutic index of aminoglycosides and improve the clinical utility of this important class of antibiotics. PMID- 11408216 TI - Novel bifunctional inhibitor of xylanase and aspartic protease: implications for inhibition of fungal growth. AB - A novel bifunctional inhibitor (ATBI) from an extremophilic Bacillus sp. exhibiting an activity against phytopathogenic fungi, including Alternaria, Aspergillus, Curvularia, Colletotricum, Fusarium, and Phomopsis species, and the saprophytic fungus Trichoderma sp. has been investigated. The 50% inhibitory concentrations of ATBI ranged from 0.30 to 5.9 microg/ml, whereas the MIC varied from 0.60 to 3.5 microg/ml for the fungal growth inhibition. The negative charge and the absence of periodic secondary structure in ATBI suggested an alternative mechanism for fungal growth inhibition. Rescue of fungal growth inhibition by the hydrolytic products of xylanase and aspartic protease indicated the involvement of these enzymes in cellular growth. The chemical modification of Asp or Glu or Lys residues of ATBI by 2,4,6-trinitrobenzenesulfonic acid and Woodward's reagent K, respectively, abolished its antifungal activity. In addition, ATBI also inhibited xylanase and aspartic protease competitively, with K(i) values 1.75 and 3.25 microM, respectively. Our discovery led us to envisage a paradigm shift in the concept of fungal growth inhibition for the role of antixylanolytic activity. Here we report for the first time a novel class of antifungal peptide, exhibiting bifunctional inhibitory activity. PMID- 11408217 TI - Influence of human serum on antifungal pharmacodynamics with Candida albicans. AB - Antifungal susceptibilities (NCCLS, approved standard M27-A, 1997) were determined for the reference strain ATCC 90028 and 21 clinical isolates of Candida albicans with varying levels of fluconazole susceptibility using RPMI 1640 (RPMI) and 80% fresh human serum-20% RPMI (serum). Sixty-four percent (14 of 22) of the isolates tested demonstrated significant decreases (> or = 4-fold) in fluconazole MICs in the presence of serum, and the remaining eight isolates exhibited no change. Itraconazole and ketoconazole, two highly protein-bound antifungal agents, had MICs in serum that were increased or unchanged for 46% (10 of 22) and 41% (9 of 22) of the isolates, respectively. All 10 isolates tested against an investigational antifungal agent, LY303366, demonstrated significant increases in the MIC required in serum, while differences in amphotericin B MICs in the two media were not observed. Four of 10 isolates tested demonstrated fourfold higher flucytosine MICs in serum than in RPMI. Postantifungal effects (PAFEs) and 24-h kill curves were determined by standard methods for selected isolates. At the MIC, fluconazole, itraconazole, ketoconazole, flucytosine, and LY303366 kill curves and PAFEs in RPMI were similar to those in serum. Isolates of fluconazole-resistant C. albicans required lower MICs in serum than in RPMI, without relative increases in fungal killing or PAFEs. Isolates tested against amphotericin B demonstrated significantly reduced killing and shorter PAFEs in serum than in RPMI without observable changes in MIC. In conclusion, antifungal pharmacodynamics in RPMI did not consistently predict antifungal activity in serum for azoles and amphotericin B. Generally speaking, antifungal agents with high protein binding exhibited some form of reduced activity (MIC, killing, or PAFE) in the presence of serum compared to those with low protein binding. PMID- 11408218 TI - Inhibition of fumarate reductase in Leishmania major and L. donovani by chalcones. AB - Our previous studies have shown that chalcones exhibit potent antileishmanial and antimalarial activities in vitro and in vivo. Preliminary studies showed that these compounds destroyed the ultrastructure of Leishmania parasite mitochondria and inhibited the respiration and the activity of mitochondrial dehydrogenases of Leishmania parasites. The present study was designed to further investigate the mechanism of action of chalcones, focusing on the parasite respiratory chain. The data show that licochalcone A inhibited the activity of fumarate reductase (FRD) in the permeabilized Leishmania major promastigote and in the parasite mitochondria, and it also inhibited solubilized FRD and a purified FRD from L. donovani. Two other chalcones, 2,4-dimethoxy-4'-allyloxychalcone (24m4ac) and 2,4 dimethoxy-4'-butoxychalcone (24mbc), also exhibited inhibitory effects on the activity of solubilized FRD in L. major promastigotes. Although licochalcone A inhibited the activities of succinate dehydrogenase (SDH), NADH dehydrogenase (NDH), and succinate- and NADH-cytochrome c reductases in the parasite mitochondria, the 50% inhibitory concentrations (IC(50)) of licochalcone A for these enzymes were at least 20 times higher than that for FRD. The IC(50) of licochalcone A for SDH and NDH in human peripheral blood mononuclear cells were at least 70 times higher than that for FRD. These findings indicate that FRD, one of the enzymes of the parasite respiratory chain, might be the specific target for the chalcones tested. Since FRD exists in the Leishmania parasite and does not exist in mammalian cells, it could be an excellent target for antiprotozoal drugs. PMID- 11408220 TI - Comparison of hydrogen peroxide contact lens disinfection systems and solutions against Acanthamoeba polyphaga. AB - Acanthamoeba is a free-living amoeba causing a potentially blinding infection of the cornea. Contact lens wearers are most at risk and account for some 95% of cases. Hydrogen peroxide is used for contact lens disinfection due to its broad antimicrobial activity. Lenses must be neutralized before use to avoid pronounced stinging and possible corneal damage. Neutralization is achieved by adding a catalyst during the disinfection process (one-step) or afterwards (two-step). Here, the activities of commercial peroxide systems and individual solutions against trophozoites and cysts of Acanthamoeba polyphaga were compared. All disinfection systems were active against trophozoites, giving a > or = 3-log (99.9%) kill within 1 h. Of the four one-step systems, only one showed some cysticidal activity, giving a 1.28 +/- 0.41-log reduction. Both two-step systems were cysticidal, giving a > or = 3-log kill at 4 h. All system peroxide solutions were cysticidal, giving a > or = 3-log kill by 4 to 6 h. Variation in the cysticidal rate was observed with two solutions that gave a 1.8- to 2.1-log kill at 4 h compared with 3.0 to 4.0 for the rest (P < 0.05). No cysticidal activity was found with the peroxigen sodium perborate or the contact lens protein remover subtilisin A. Two-step systems are cysticidal providing contact times of at least 4 h are employed. Variation in cyst killing occurs between peroxide solutions, possibly due to formulation differences. One-step systems are less effective against Acanthamoeba cysts due to rapid peroxide neutralization. The cysticidal activity of one-step systems could be improved if neutralization rates were retarded. PMID- 11408219 TI - Identification of Proteus mirabilis mutants with increased sensitivity to antimicrobial peptides. AB - Antimicrobial peptides (APs) are important components of the innate defenses of animals, plants, and microorganisms. However, some bacterial pathogens are resistant to the action of APs. For example, Proteus mirabilis is highly resistant to the action of APs, such as polymyxin B (PM), protegrin, and the synthetic protegrin analog IB-367. To better understand this resistance, a transposon mutagenesis approach was used to generate P. mirabilis mutants sensitive to APs. Four unique PM-sensitive mutants of P. mirabilis were identified (these mutants were >2 to >128 times more sensitive than the wild type). Two of these mutants were also sensitive to IB-367 (16 and 128 times more sensitive than the wild type). Lipopolysaccharide (LPS) profiles of the PM- and protegrin-sensitive mutants demonstrated marked differences in both the lipid A and O-antigen regions, while the PM-sensitive mutants appeared to have alterations of either lipid A or O antigen. Matrix-assisted laser desorption ionization-time of flight mass spectrometry analysis of the wild-type and PM sensitive mutant lipid A showed species with one or two aminoarabinose groups, while lipid A from the PM- and protegrin-sensitive mutants was devoid of aminoarabinose. When the mutants were streaked on an agar-containing medium, the swarming motility of the PM- and protegrin-sensitive mutants was completely inhibited and the swarming motility of the mutants sensitive to only PM was markedly decreased. DNA sequence analysis of the mutagenized loci revealed similarities to an O-acetyltransferase (PM and protegrin sensitive) and ATP synthase and sap loci (PM sensitive). These data further support the role of LPS modifications as an elaborate mechanism in the resistance of certain bacterial species to APs and suggest that LPS surface charge alterations may play a role in P. mirabilis swarming motility. PMID- 11408221 TI - Effect of famciclovir on herpes simplex virus type 1 corneal disease and establishment of latency in rabbits. AB - Famciclovir (FCV) is efficacious in the treatment of acute herpes zoster and recurrent genital infections but has not been used to treat ocular herpes simplex virus (HSV) infections. We evaluated the efficacy of orally administered FCV in treating HSV-1 epithelial keratitis and determined its effects on the establishment of latency and subsequent reactivation. Rabbits were inoculated with HSV-1 strain 17 syn+ and treated twice daily with increasing concentrations of FCV (60 to 500 mg/kg of body weight). This resulted in a significant, dose dependent improvement in keratitis scores, as well as prolonged survival. Regardless of the dose of drug used, all groups exhibited the high rates of spontaneous and induced reactivation characteristic of 17syn+. The efficacy of 250 mg of FCV per kg was also compared to topical treatment with 1% trifluorothymidine (TFT). Although TFT treatment was more effective at reducing eye disease, FCV-treated rabbits had a better survival rate. Real-time quantitative PCR analysis of rabbit trigeminal ganglia (TG) demonstrated that FCV significantly reduced the HSV-1 copy number compared to that after treatment with TFT or the placebo but not in a dose-dependent manner. In summary, oral FCV treatment significantly reduces the severity of corneal lesions, reduces the number of HSV-1 genomes in the TG, improves survival, and therefore may be beneficial in reducing the morbidity of HSV keratitis in the clinic. PMID- 11408222 TI - Effect of abolishment of the use of antimicrobial agents for growth promotion on occurrence of antimicrobial resistance in fecal enterococci from food animals in Denmark. AB - From 1995 to 2000, a total of 673 Enterococcus faecium and 1,088 Enterococcus faecalis isolates from pigs together with 856 E. faecium isolates from broilers were isolated and tested for susceptibility to four classes of antimicrobial agents used for growth promotion as part of the Danish program of monitoring for antimicrobial resistance. The four antimicrobials were avilamycin, erythromycin, vancomycin, and virginiamycin. Major changes in the use of antimicrobial agents for growth promotion have occurred during the last 6 years in Denmark. The government banned the use of avoparcin in 1995 and of virginiamycin in 1998. Furthermore, the producers have voluntarily stopped all use beginning in 1999. The avoparcin ban in 1995 was followed by a decrease in the occurrence of glycopeptide-resistant E. faecium (GRE) in broilers, from 72.7% in 1995 to 5.8% in 2000. The occurrence of glycopeptide resistance among isolates from pigs remained constant at around 20% from 1995 to 1997. It was shown that, in GRE from pigs, the genes encoding macrolide and glycopeptide resistance were genetically linked and that, following the decrease in the use of tylosin during 1998 and 1999, the occurrence of GRE in pigs decreased to 6.0% in 2000. From 1995 to 1997 the occurrence of erythromycin resistance among E. faecium and E. faecalis isolates from pigs was almost 90%. Use of tylosin decreased considerably during 1998 and 1999, and this decrease was followed by decreases in the occurrence of resistance to 46.7 and 28.1% among E. faecium and E. faecalis isolates from pigs, respectively. Erythromycin resistance among E. faecium isolates from broilers reached a maximum of 76.3% in 1997 but decreased to 12.7% in 2000 concomitantly with more limited use of virginiamycin. Use of virginiamycin increased from 1995 to 1997 and was followed by an increased occurrence of virginiamycin resistance among E. faecium isolates in broilers, from 27.3% in 1995 to 66.2% in 1997. In January 1998 the use of virginiamycin was banned in Denmark, and the occurrence of virginiamycin resistance decreased to 33.9% in 2000. Use of avilamycin increased from 1995 to 1996 and was followed by an increase in avilamycin resistance among E. faecium isolates from broilers, from 63.6% in 1995 to 77.4% in 1996. Since 1996 avilamycin usage has decreased, followed by a decrease in resistance to 4.8% in 2000. Our observations show that it is possible to reduce the occurrence of antimicrobial resistance in a national population of food animals when the selective pressure is removed. Cases in which resistance to vancomycin was linked to resistance to erythromycin were exceptions. In such cases resistance did not decrease until the use of both avoparcin and tylosin was limited. PMID- 11408223 TI - Heat treatment of amphotericin b modifies its serum pharmacokinetics, tissue distribution, and renal toxicity following administration of a single intravenous dose to rabbits. AB - The purpose of this investigation was to determine the serum pharmacokinetics, tissue distribution, and renal toxicity of amphotericin B (AmpB) following administration of a single intravenous dose (1 mg/kg of body weight) of Fungizone (FZ) and a heat-treated form of FZ (HFZ) to New Zealand White female rabbits. FZ solutions were heated at 70 degrees C for 20 min to produce HFZ. Blood samples were obtained before drug administration and serially thereafter. After collection of the 48-h blood sample, each rabbit was humanely sacrificed and the right kidney, spleen, lungs, liver, and heart were harvested for AmpB analysis. Serum creatinine levels were measured before and 10 h after drug administration. AmpB concentrations in the serum and tissues were analyzed using high-performance liquid chromatography. FZ administration to rabbits resulted in a greater-than 50% increase in serum creatinine concentrations compared to baseline. However, HFZ administration resulted in no difference in serum creatinine concentrations compared to baseline. The AmpB area under the concentration-time curve (AUC) after HFZ administration was significantly lower than the AmpB AUC in rabbits administered FZ. However, AmpB systemic total body clearance was significantly greater in rabbits administered HFZ than in rabbits administered FZ without any differences in volume of distribution at steady state. Kidney tissue AmpB concentrations, although not significantly different, were greater in rabbits administered FZ than in rabbits administered HFZ. Likewise, lung and spleen AmpB concentrations, although not significantly different, were greater in rabbits administered FZ than in rabbits administered HFZ. However, liver AmpB concentrations were significantly lower in rabbits administered FZ than in rabbits administered HFZ. No significant differences in heart AmpB concentration between rabbits administered FZ and those given HFZ were found. These findings suggest that the pharmacokinetics, tissue distribution, and renal toxicity of AmpB are modified following administration of HFZ. HFZ could be an improved low cost AmpB drug delivery system that has a potentially higher therapeutic index than FZ. PMID- 11408224 TI - Antimonial-mediated DNA fragmentation in Leishmania infantum amastigotes. AB - The basic treatment of leishmaniasis consists in the administration of pentavalent antimonials. The mechanisms that contribute to pentavalent antimonial toxicity against the intracellular stage of the parasite (i.e., amastigote) are still unknown. In this study, the combined use of several techniques including DNA fragmentation assay and in situ and cytofluorometry terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling methods and YOPRO-1 staining allowed us to demonstrate that potassium antimonyl tartrate, an Sb(III)-containing drug, was able to induce cell death associated with DNA fragmentation in axenic amastigotes of Leishmania infantum at low concentrations (10 microg/ml). This observation was in close correlation with the toxicity of Sb(III) species against axenic amastigotes (50% inhibitory concentration of 4.75 microg/ml). Despite some similarities to apoptosis, nuclease activation was not a consequence of caspase-1, caspase-3, calpain, cysteine protease, or proteasome activation. Altogether, our results demonstrate that the antileishmanial toxicity of Sb(III) antimonials is associated with parasite oligonucleosomal DNA fragmentation, indicative of the occurrence of late events in the overall process of apoptosis. The elucidation of the biochemical pathways leading to cell death could allow the isolation of new therapeutic targets. PMID- 11408225 TI - Detection of resistance to amphotericin B in Candida isolates by using Iso Sensitest broth. AB - A major limitation of the National Committee for Clinical Laboratory Standards M27-A methodology is reliable detection of amphotericin B (AMB) resistance. The results obtained by using Iso-Sensitest, a synthetic medium, to detect AMB resistance were analyzed and compared with those obtained with RPMI and antibiotic medium 3 (AM3). The ability to detect AMB resistance with RPMI is not enhanced by using a higher inoculum, glucose supplementation at a final concentration of 20 g/liter, spectrophotometric reading, or 24 h of incubation time. Testing using AM3 and an inoculum of 10(3) CFU/ml detects resistance. Identification of resistant isolates is not improved by glucose supplementation, changes in reading method, or changes in incubation time. However, the use of AM3 as assay medium and an inoculum of 10(5) CFU/ml did not allow detection of AMB resistance. Testing using Iso-Sensitest medium appears to be similar to AM3 in detecting resistance. The most pronounced discrimination is achieved by testing in Iso-Sensitest supplemented with glucose and spectrophotometric reading after 24 h of incubation. The reproducibility of MIC testing was greatest for Iso Sensitest-based procedures. Use of Iso-Sensitest produces both highly reproducible MICs and reliable identification of AMB-resistant Candida isolates. PMID- 11408226 TI - All detectable high-molecular-mass penicillin-binding proteins are modified in a high-level beta-lactam-resistant clinical isolate of Streptococcus mitis. AB - All detectable high-molecular-mass penicillin-binding proteins (HMM PBPs) are altered in a clinical isolate of Streptococcus mitis for which the beta-lactam MICs are increased from those previously reported in our region (cefotaxime MIC, 64 microg/ml). These proteins were hardly detected at concentrations that saturate all PBPs in clinical isolates and showed, after densitometric analysis, 50-fold-lower radiotracer binding. Resistance was related to mosaic structure in all HMM PBP-coding genes, where critical region replacement was complemented not only by substitutions already reported for the closely related Streptococcus pneumoniae but also by other specific replacements that are presumably close to the active-site serine. Mosaic structure was also presumed in a pbp1a-sensitive strain used for comparison, confirming that these structures do not unambiguously imply, by themselves, detectable critical changes in the kinetic properties of these proteins. PMID- 11408228 TI - Pharmacodynamic assessment of gatifloxacin against Streptococcus pneumoniae. AB - The pharmacodynamic parameters of peak serum drug concentration/MIC (peak/MIC) ratio and the area under the curve (AUC)/MIC ratio have been used to characterize in vivo drug exposure and its relationship to bacterial killing for the fluoroquinolones. Our study objectives were to describe the pharmacodynamic relationship between gatifloxacin exposure and outcome as assessed by bacterial density and survival in an immunocompromised murine thigh model of pneumococcal infection and to assess the relationship between drug exposure and these outcomes in an immunocompetent host. ICR mice were rendered neutropenic, and thigh infection was induced by intramuscular administration of 0.1 ml of 10(5) to 10(7) CFU of Streptococcus pneumoniae/ml. Mice received 1 to 5 mg of uranyl nitrate/kg of body weight at day -3 and were randomized to receive 10 to 80 mg of gatifloxacin/kg every 6 to 24 h orally, starting at 2 h postinoculation. Bacterial density studies were completed 24 h after initiation of therapy, and survival was assessed after 4 days of treatment. MICs for clinical isolates (n = 8) ranged from 0.25 to 1.0 microg/ml. Correlations were assessed between the change in bacterial density, as well as survival, and the AUC/MIC ratio, peak/MIC ratio, and the duration of time that serum drug concentration remained above the MIC. The best predictor of bacterial response was the AUC/MIC ratio for both outcome measures. There was greater efficacy, as measured by a decrease in log change in CFU as well as by survival data, in the immunocompetent mice compared to the immunocompromised mice. These data demonstrate (i) the appropriateness of the AUC/MIC ratio as a dynamic predictor of response to pneumococcal infection for the fluoroquinolones, (ii) that gatifloxacin AUC/MIC ratios of 30 to 40 appear to optimize bactericidal activity and survival in this model, and (iii) that immunocompetency of the host plays a role in efficacy. PMID- 11408227 TI - Induction of Epstein-Barr virus kinases to sensitize tumor cells to nucleoside analogues. AB - The presence of Epstein-Barr virus (EBV) in the tumor cells of some EBV associated malignancies may facilitate selective killing of these tumor cells. We show that treatment of an EBV(+) Burkitt's lymphoma cell line with 5-azacytidine led to a dose-dependent induction of EBV lytic antigen expression, including expression of the viral thymidine kinase (TK) and phosphotransferase (PT). Azacytidine treatment for 24 h modestly sensitized the cell line to all nucleosides tested. To better characterize EBV TK with regard to various nucleoside analogues, we expressed EBV TK in stable cell clones. Two EBV TK expressing clones were moderately sensitive to high doses of acyclovir and penciclovir (PCV) (62.5 to 500 microM) and to lower doses of ganciclovir (GCV) and bromovinyldeoxyuridine (BVdU) (10 to 100 microM) compared to a control clone and were shown to phosphorylate GCV. Similar experiments in a transient overexpression system showed more killing of cells transfected with the EBV TK expression vector than of cells transfected with the control mutant vector (50 microM GCV for 4 days). A putative PT was also studied in the transient transfection system and appeared similar to the TK in phosphorylating GCV and conferring sensitivity to GCV, but not in BVdU- or PCV-mediated cell killing. Induction of EBV kinases in combination with agents such as GCV merits further evaluation as an alternative strategy to gene therapy for selective killing of EBV-infected cells. PMID- 11408229 TI - DNA gyrase-mediated natural resistance to fluoroquinolones in Ehrlichia spp. AB - Fluoroquinolone susceptibility heterogeneity between various Ehrlichia species has been previously demonstrated. In gram-negative bacteria, resistance to fluoroquinolones most often corresponds to specific amino acid variations in a portion of the protein sequence of the A subunit of DNA gyrase (GyrA), referred to as the quinolone resistance-determining region (QRDR). We suspected a similar mechanism to be responsible for natural resistance in some Ehrlichia species. To verify this hypothesis, we sequenced the entire gyrA gene of the quinolone susceptible species Ehrlichia sennetsu and designed specific primers to amplify and sequence the QRDR of four other Ehrlichia species as well as the closely related species Cowdria ruminantium. We identified in the fluoroquinolone resistant species Ehrlichia chaffeensis and Ehrlichia canis a specific GyrA QRDR amino acid sequence, also present in C. ruminantium (whose susceptibility to fluoroquinolones remains unknown). These three species belong to a single phylogenetic cluster referred to as the E. canis genogroup. A different GyrA QRDR pattern, shared by the Ehrlichia species representatives of the E. sennetsu and Ehrlichia phagocytophila genogroups, was identified. Three of the four species tested are known to be susceptible to fluoroquinolones. A serine residue in position 83 (Escherichia coli numbering) in the susceptible species is replaced by an alanine residue in fluoroquinolone-resistant species. These results are consistent with the current knowledge on fluoroquinolone resistance in other gram negative bacteria. They are indicative of a natural gyrase-mediated resistance to fluoroquinolones in the E. canis genogroup. PMID- 11408230 TI - Desbutyl-benflumetol, a novel antimalarial compound: in vitro activity in fresh isolates of Plasmodium falciparum from Thailand. AB - Desbutyl-benflumetol (DBB) is a novel antimalarial compound closely related to benflumetol (lumefantrine), of which it is a putative metabolite. The in vitro response of Plasmodium falciparum to DBB was studied in Mae Hong Son and Mae Sot, in northwest Thailand, in 1997 and 1998. In total, 155 fresh isolates were successfully tested using the World Health Organization standard in vitro microtest system (Mark II). The mean 50% effective concentration (EC(50)) and 90% effective concentration of DBB were 6.36 and 31.09 nmol/liter, respectively. The comparison of the activity of DBB and benflumetol yielded a highly significant potency ratio of 4.52, corresponding to a more than four times higher efficacy of DBB. A considerable potency difference was found between isolates from Mae Hong Son and those from Mae Sot, reflecting lesser sensitivity in the area with marked resistance to mefloquine and quinine. This observation is also supported by a highly significant activity correlation with benflumetol (P < 0.001) and to a similar degree with mefloquine (P < 0.001), reflecting a close relationship of DBB with the class II aryl amino alcohol blood schizontocides. A less distinct association was also found with artemisinin, which was significant only at the EC(50) level, and there was no correlation at all with chloroquine. DBB is a promising antimalarial compound that merits further investigation in order to define its practical therapeutic potential. PMID- 11408231 TI - Discrimination of SHV beta-lactamase genes by restriction site insertion-PCR. AB - Restriction site insertion-PCR (RSI-PCR) is a simple, rapid technique for detection of point mutations. This technique exploits primers with one to three base mismatches near the 3' end to modulate a restriction site. We have developed this technique to identify described mutations of the bla(SHV) genes for differentiation of SHV variants that cannot be distinguished easily by other techniques. To validate this method, eight standard strains were used, each producing a different SHV beta-lactamase: SHV-1, SHV-2, SHV-3, SHV-4, SHV-5, SHV 6, SHV-8, and SHV-18. Mismatch primers were designed to detect mutations affecting amino acids at positions 8 (SspI), 179 (HinfI), 205 (PstI), 238 (Gly- >Ala) (BsrI), and 240 (NruI) of bla(SHV) genes. All amplimers of the bla(SHV) genes used in this study yielded the predicted restriction endonuclease digestion products. In addition, this study also makes theoretical identification of bla(SHV-6), bla(SHV-8), and 12 novel bla(SHV) variants using the PCR-restriction fragment length polymorphism (RFLP) technique possible. By using a combination of PCR-RFLP and RSI-PCR techniques, up to 27 SHV variants can now be distinguished rapidly and reliably. These simple techniques are readily applied to epidemiological studies of the SHV beta-lactamases and may be extended to the characterisation of other resistance determinants. PMID- 11408232 TI - Pharmacodynamic evaluation of RWJ-270201, a novel neuraminidase inhibitor, in a lethal murine model of influenza predicts efficacy for once-daily dosing. AB - We examined RWJ-270201 in a lethal model of influenza in BALB/c mice. The aim was to delineate the pharmacodynamically linked variable for the drug. Challenge was performed with influenza virus A/Shongdong/09/93 (H3N2). Treatment was administered by gavage. Five doses (1 to 10 mg/kg of body weight) and three schedules (every 24, 12, and 8 h) were evaluated with 10 mice per group. There were 39 placebo-treated mice. Drug exposure was evaluated for infected mice. Exposures were calculated after population modeling of all the plasma concentration-time data simultaneously using the NPEM3 program. Evaluation of dose and schedule with Kaplan-Meier analysis and Cox proportional hazards modeling demonstrated that schedule offered no explanatory power relative to dose alone. Evaluation of peak concentration, trough concentration, and area under the concentration-time curve (AUC) by the same methods revealed that AUC was the dynamically linked variable. Again, schedule offered no further explanatory power when included in the model with AUC. This indicates that AUC is the linked variable and that the anti-influenza effect of RWJ-270201 is independent of schedule. We conclude that once-daily dosing of RWJ-270201 should be evaluated in clinical trials of influenza therapy. PMID- 11408233 TI - In vitro antibiotic susceptibilities of Burkholderia mallei (causative agent of glanders) determined by broth microdilution and E-test. AB - In vitro susceptibilities to 28 antibiotics were determined for 11 strains of Burkholderia mallei by the broth microdilution method. The B. mallei strains demonstrated susceptibility to aminoglycosides, macrolides, quinolones, doxycycline, piperacillin, ceftazidime, and imipenem. For comparison and evaluation, 17 antibiotic susceptibilities were also determined by the E-test. E test values were always lower than the broth dilution values. Establishing and comparing antibiotic susceptibilities of specific B. mallei strains will provide reference information for assessing new antibiotic agents. PMID- 11408234 TI - Safety and pharmacokinetics of multiple 750-milligram doses of intravenous levofloxacin in healthy volunteers. AB - The safety and pharmacokinetics of a once-daily high intravenous dose of levofloxacin (750 mg) in 18 healthy volunteers were studied in a double-blind, randomized, placebo-controlled, single-center parallel group study. Levofloxacin was well tolerated, and higher maximum concentration of drug in serum and area under the concentration-time curve values were achieved. For difficult-to-treat infections, high daily doses of levofloxacin may be beneficial, and intravenous administration may be preferred in certain clinical settings, such as when treating patients in intensive care units, warranting further evaluation. PMID- 11408235 TI - Antimicrobial activities of gatifloxacin against nosocomial isolates of Stenotrophomonas maltophilia measured by MIC and time-kill studies. AB - We determined in vitro activities of gatifloxacin and seven other drugs against 100 isolates of Stenotrophomonas maltophilia using the agar gradient diffusion (Etest) method. Percentages of susceptible isolates were as follows: trimethoprim sulfamethoxazole, 90%; gatifloxacin, 71%; levofloxacin, 57%; ticarcillin clavulanic acid, 54%; ceftazidime, 49%; ciprofloxacin, 29%; cefepime, 21%; and piperacillin-tazobactam, 20%. Time-kill studies of three isolates indicated that gatifloxacin was bactericidal at times as early as 3 h of incubation when tested at concentrations equivalent to twice the MIC (two isolates) and 4 times the MIC (one isolate). PMID- 11408236 TI - Prospective study of Candida species in patients at a comprehensive cancer center. AB - Since most nosocomial systemic yeast infections arise from the endogenous flora of the patient, we prospectively evaluated the species stratification and antifungal susceptibility profile of Candida spp. associated with heavy colonization and systemic infection in patients at Memorial Sloan-Kettering Cancer Center in New York. A total of 349 Candida isolates were obtained from 223 patients during the later half of 1998. Cancer was the most common underlying disease, occurring in 91% of the patients, including 61.8% with organ and 23.7% with hematological malignancies; 4.4% of the patients had AIDS. Candida albicans was the predominant species (67.3%); among 114 non-albicans Candida spp., C. glabrata (45.6%) was the most frequent, followed by C. tropicalis (18.4%), C. parapsilosis (16.6%), and C. krusei (9.6%). The overall resistance to triazole based agents among all yeast isolates was 9.4 and 10.8% for fluconazole and itraconazole, respectively. A total of 5% of C. albicans strains were resistant to triazole antifungals, whereas 30.8 and 46.2% of C. glabrata strains were resistant to fluconazole (MIC > or = 64 microg/ml) and itraconazole (MIC > or = 1 microg/ml), respectively. A significant association was observed between prior treatment with triazole and isolation of fluconazole-resistant C. albicans (P = 0.005, OR 36), although this relationship was not seen in C. glabrata isolates (P = 0.4). This study reinforces the importance of periodic, prospective surveillance of clinical fungal isolates to determine appropriate prophylactic, empiric, and preemptive antifungal therapy for the highly susceptible patient population. PMID- 11408237 TI - Clarithromycin resistance and eradication of Helicobacter pylori in children. AB - Outcome of Helicobacter pylori infection was analyzed in 61 children treated with a triple therapy including clarithromycin. Bacterial eradication was obtained in all children with clarithromycin-susceptible strains but not in children with clarithromycin-resistant ones (P = 0.0001). H. pylori antimicrobial susceptibility is mandatory before choosing a treatment, and clarithromycin should be avoided in case of resistance. PMID- 11408238 TI - Fluoroquinolone resistance in anaerobic bacteria following exposure to levofloxacin, trovafloxacin, and sparfloxacin in an in vitro pharmacodynamic model. AB - This investigation explored pharmacodynamic characteristics of fluoroquinolones against Bacteroides thetaiotamicron and the potential for development of resistance. An in vitro model was used to generate kill curves with three fluoroquinolones at various area under the concentration-time curve (AUC)/MIC ratios. Concentration-independent killing was observed. Increases in MICs were noted following exposure to fluoroquinolones at AUC/MIC ratios of 6 to 14. PMID- 11408239 TI - Molecular characterization of a novel plasmid-encoded cefotaximase (CTX-M-12) found in clinical Klebsiella pneumoniae isolates from Kenya. AB - Nine Klebsiella pneumoniae isolates, six from blood and three from cerebrospinal fluid of newborn babies at Kenyatta National Hospital, Nairobi, Kenya, were analyzed for the mechanism of cephalosporin resistance. By using pulsed-field gel electrophoresis of XbaI-digested chromosomal DNA, all the nine isolates were found to be clonal. PCR and direct sequencing revealed a novel extended-spectrum beta-lactamase, which we designated CTX-M-12. It has a more potent hydrolytic activity against cefotaxime than against ceftazidime and a pI of 9.0 and is encoded on a large self-transferable ca. 160-kbp plasmid. PMID- 11408240 TI - Biochemical mechanism of human immunodeficiency virus type 1 reverse transcriptase resistance to stavudine. AB - We have found a close correlation between viral stavudine (d4T) resistance and resistance to d4T-triphosphate at the human immunodeficiency virus type 1 reverse transcriptase (RT) level. RT from site-directed mutants with 69S-XX codon insertions and/or conventional zidovudine resistance mutations seems to be involved in an ATP-dependent resistance mechanism analogous to pyrophosphorolysis, whereas the mechanism for RT with the Q151M or V75T mutation appears to be independent of added ATP for reducing binding to d4T-triphosphate. PMID- 11408241 TI - Macrolide-resistant Streptococcus pneumoniae in Canada during 1998-1999: prevalence of mef(A) and erm(B) and susceptibilities to ketolides. AB - In this study (1998-1999), we collected 215 macrolide-resistant Streptococcus pneumoniae isolates from an ongoing Canadian Respiratory Organism Surveillance Study involving 23 centers representing all regions of Canada. The prevalence of erythromycin-resistant S. pneumoniae was 8% (215 of 2,688). Of the 215 isolates, 48.8% (105 of 215) were PCR positive for mef(A) and 46.5% (100 of 215) were PCR positive for erm(B). The ketolides telithromycin and ABT-773 demonstrated excellent activity against both mef(A) (MIC for 90% of strains [MIC(90)], 0.06 and 0.03 microg/ml, respectively) and erm(B) (MIC(90), 0.06 and 0.03 microg/ml, respectively) strains of S. pneumoniae. PMID- 11408242 TI - In vitro activities of four novel triazoles against Scedosporium spp. AB - In order to develop new approaches to the treatment of the severe and usually fatal infections caused by Scedosporium spp., the in vitro antifungal activities of four novel triazoles (posaconazole, ravuconazole, voriconazole, and UR-9825) and some current antifungals (amphotericin B, ketoconazole, itraconazole, and nystatin) were determined. The latter group was clearly ineffective against the two species tested. The four new antifungals showed activity against Scedosporium apiospermum, and UR-9825 and voriconazole were active against S. prolificans. PMID- 11408243 TI - Resistance to linezolid: characterization of mutations in rRNA and comparison of their occurrences in vancomycin-resistant enterococci. AB - To assess the potential for emergence of resistance during the use of linezolid, we tested 10 clinical isolates of vancomycin-resistant enterococci (VRE) (four Enterococcus faecalis, five Enterococcus faecium, and one Enterococcus gallinarum) as well as a vancomycin-susceptible control (ATCC 29212) strain of E. faecalis. The enterococci were exposed to doubling dilutions of linezolid for 12 passes. After the final passage, the linezolid plate growing VRE contained a higher drug concentration with E. faecalis than with E. faecium. DNA sequencing of the 23S rRNA genes revealed that linezolid resistance in three E. faecalis isolates was associated with a guanine to uracil transversion at bp 2576, while the one E. faecium isolate for which the MIC was 16 microg/ml contained a guanine to adenine transition at bp 2505. PMID- 11408244 TI - Novel mutations in ndh in isoniazid-resistant Mycobacterium tuberculosis isolates. AB - Novel mutations in NADH dehydrogenase (ndh) were detected in 8 of 84 (9.5%) isoniazid (INH)-resistant isolates (T110A [n = 1], R268H [n = 7]), but not in 22 INH-susceptible isolates of Mycobacterium tuberculosis. Significantly, all eight isolates with mutations at ndh did not have mutations at katG, kasA, or the promoter regions of inhA or ahpC, except for one isolate. Mutations in ndh appear to be an additional molecular mechanism for isoniazid resistance in M. tuberculosis. PMID- 11408245 TI - Pharmacokinetic evaluation of oral levofloxacin in human immunodeficiency virus infected subjects receiving concomitant antiretroviral therapy. AB - The purpose of this study was to evaluate the pharmacokinetics (PK) profile of oral levofloxacin in human immunodeficiency virus-positive patients in steady state treatment with nelfinavir (NFV) or with efavirenz (EFV) and to determine the effects of levofloxacin on the PK parameters of these two antiretroviral agents. For levofloxacin, plasma samples were obtained at steady state during a 24-h dosing interval. Plasma NFV and EFV concentrations were evaluated before and after 4 days of levofloxacin treatment. Levofloxacin PK do not seem affected by NFV and EFV. There was no significant difference between NFV and EFV plasma levels obtained with and without levofloxacin. PMID- 11408246 TI - Comparative in vitro activity of ABT-773, a novel antibacterial ketolide. AB - The in vitro activities of ABT-773, erythromycin, clarithromycin, and azithromycin were compared. ABT-773 was the most active compound against macrolide-susceptible Streptococcus pneumoniae, Streptococcus pyogenes, Staphylococcus aureus, Staphylococcus epidermidis, Listeria monocytogenes, and Enterococcus spp. and multidrug-resistant Streptococcus pneumoniae. It also had good activity against gram-negative and atypical respiratory tract pathogens and Helicobacter pylori. PMID- 11408247 TI - Activity of LY333328 in experimental meningitis caused by a Streptococcus pneumoniae strain susceptible to penicillin. AB - In a rabbit model of Streptococcus pneumoniae meningitis single doses of 10 and 2.5 mg of the glycopeptide LY333328 per kg of body weight reduced bacterial titers in cerebrospinal fluid (CSF) almost as rapidly as ceftriaxone at 10 mg/kg/h (changes in log CFU, -0.29 +/- 0.21 and -0.26 +/- 0.22 versus -0.34 +/- 0.15/ml/h). A dose of 1 mg/kg was bacteriostatic (change in log CFU, 0.01 +/- 0.11/ml/h). In two animals receiving LY333328 at a dose of 40 mg/kg the bacterial titers were reduced by 0.54 and 0.51 log CFU/ml/h. The penetration of CSF by LY333328 was 1 to 5%. The concentrations of lipoteichoic and teichoic acids in CSF and neuronal damage were similar in ceftriaxone- and LY333328-treated animals. PMID- 11408248 TI - Domestic cat model for predicting human nucleoside analogue pharmacokinetics in blood and seminal plasma. AB - To establish whether a feline model can predict nucleoside analogue behavior in human semen, zidovudine (ZDV) and lamivudine (3TC) pharmacokinetic parameters (PKs) were determined in the blood and seminal plasma of healthy cats. Our results show considerable similarity in ZDV and 3TC PKs between cats and humans. As in humans, ZDV and 3TC tend to accumulate in feline seminal plasma. Area under the blood plasma concentration-time curve was predictive of seminal plasma excretion. The felid model offers a unique in vivo experimental alternative for investigating the pharmacokinetics of nucleoside analogues in the male genital tract. PMID- 11408249 TI - Microbes and microbial toxins: paradigms for microbial-mucosal interactions. VII. Enteropathogenic Escherichia coli: physiological alterations from an extracellular position. AB - Enteropathogenic Escherichia coli (EPEC) is primarily associated with infantile diarrhea in developing countries. This intriguing pathogen exerts numerous physiological effects on its host target tissue, the intestinal epithelium, all from an extracellular location. Expression of a type III secretory apparatus allows this organism to transfer bacterial effector molecules directly into host cells. As a result of EPEC attachment to and/or translocation of proteins into intestinal epithelial cells, many signaling cascades are activated. Ultimately, host functions are perturbed, including alteration of ion transport, disruption of the tight junction barrier, and activation of the inflammatory response. PMID- 11408250 TI - Receptors and transmission in the brain-gut axis: potential for novel therapies. III. Mu-opioid receptors in the enteric nervous system. AB - G protein-coupled receptors are cell surface signal-transducing proteins, which elicit a variety of biological functions by the activation of different intracellular effector systems. Many of these receptors, including the mu-opioid receptor (mu OR), have been localized in the gastrointestinal tract. mu OR is the target of opioids and alkaloids, potent analgesic drugs with high potential for abuse. mu OR is expressed by enteric neurons, and it undergoes ligand-selective endocytosis. It is of clinical importance because it mediates tolerance and other major side effects of opiate analgesics, including impairment of gastrointestinal propulsion. An important observation of mu OR is its differential trafficking and desensitization properties in response to individual agonists, which might have long-term physiological consequences and be involved in the development of opiate side effects. Receptor activation by agonists is the basis for signaling, and alterations of the mechanisms controlling cellular responses of G protein-coupled receptors to agonists might be the basis of several diseases, including gastrointestinal diseases. Therefore, understanding these basic cellular mechanisms is important for developing appropriate therapeutic agents. PMID- 11408251 TI - Inhibition of gastrointestinal lipolysis by Orlistat during digestion of test meals in healthy volunteers. AB - The inhibition of digestive lipases by the antiobesity drug Orlistat along with lipolysis levels and fecal fat excretion were measured in healthy humans. Orlistat was found to be a powerful gastric lipase inhibitor, achieving 46.6- 91.4% enzyme inhibition and thus greatly reducing gastric lipolysis of solid and liquid meals (11--33% of respective controls). Gastric lipase inhibition by Orlistat was extremely fast (half-inhibition time < 1 min). Duodenal lipolysis was reduced significantly by Orlistat given with the solid meal (32.6--37.6% of controls) but was only slightly reduced by Orlistat given with the liquid meal (74.5--100% of controls). Human pancreatic lipase (HPL) inhibition was found to be high (51.2--82.6%), however, regardless of the meal. These paradoxical results were explained when in vitro lipolysis experiments were performed. The rates of HPL inhibition by Orlistat were found to be similar with both types of meals (half-inhibition time 5--6 min), but the preemulsified triglycerides of the liquid meal were rapidly hydrolyzed by HPL before the enzyme was significantly inhibited by Orlistat. With the solid meal, the rate of hydrolysis of the meal triglycerides by HPL was slower than the rate of HPL inhibition by Orlistat. As predicted from the previous results, the effects of Orlistat on fat excretion levels were found to be much greater with the solid (40.5--57.4% of ingested fat) than with the liquid (4.2--18.8%) test meal. PMID- 11408252 TI - Hepatic parasympathetic (HISS) control of insulin sensitivity determined by feeding and fasting. AB - In response to insulin, a hormone [hepatic insulin sensitizing substance (HISS)] is released from the liver to stimulate glucose uptake in skeletal muscle but not liver or gut. The aim was to characterize dynamic control of HISS action in response to insulin and regulation of release by hepatic parasympathetic nerves. Insulin action was assessed by the rapid insulin sensitivity test, where the index is the glucose required (mg/kg) to maintain euglycemia after a bolus of insulin. Blocking HISS release by interruption of the hepatic parasympathetic nerves by surgical denervation, atropine, or blockade of hepatic nitric oxide synthase produced similar degrees of insulin resistance and revealed a similar dynamic pattern of hormone action that began 3--4 min after, and continued for 9- 10 min beyond, insulin action (50 mU/kg). HISS action accounted for 56.5 +/- 3.5% of insulin action at insulin doses from 5 to 100 mU/kg (fed). We also tested the hypothesis that HISS release is controlled by the feed/fast status. Feeding resulted in maximal HISS action, which decreased progressively with the duration of fasting. PMID- 11408253 TI - Polyamine depletion arrests growth of IEC-6 and Caco-2 cells by different mechanisms. AB - The polyamines spermidine and spermine and their precursor, putrescine, are required for the growth and proliferation of eukaryotic cells. This study compares and contrasts growth arrest caused by polyamine depletion in the untransformed IEC-6 cell line with that in the p53-mutated colon cancer Caco-2 cell line. Cells were grown in the presence or absence of alpha difluoromethylornithine (DFMO), a specific inhibitor of ornithine decarboxylase, the first rate-limiting enzyme in the synthesis of polyamines. Depletion of polyamines inhibited the growth of both cell lines equally and over the same time frame. However, whereas IEC-6 cells were arrested in the G(1) phase of the cell cycle, there was no accumulation of Caco-2 cells in any particular phase. In IEC 6 cells, growth arrest was accompanied by elevated levels of p53 and p21(Waf1/Cip1) (p21). There were no changes in p53 levels in Caco-2 cells. Levels of p21 increased in Caco-2 cells on day 2 without any effect on cell cycle progression. The amount of cyclin-dependent kinase (Cdk)2 protein was unchanged by polyamine depletion in both cell lines. However, the activity of Cdk2 was significantly inhibited by DFMO in IEC-6 cells. These data suggest that in the untransformed IEC-6 cells the regulation of Cdk2 activity and progression through the cell cycle are p53- and p21 dependent. Growth arrest in the p53-mutated Caco 2 line after polyamine depletion occurs by a different, yet unknown, mechanism. PMID- 11408254 TI - Vagal afferent input determines the volume dependence of rat esophageal motility patterns. AB - The volume dependence of balloon distension-evoked esophageal rhythmic motor responses and their neural correlates was investigated in 72 urethane anesthetized rats. With increasing balloon volume (75--200 microl), distal esophageal rhythmic contractions decreased in rate and became tonic in the range of 150--250 microl. This change in motor pattern involved only the striated musculature of the esophageal body and persisted after acute transection of the spinal cord at C(2). Impulse frequency in single vagal afferents of the distal esophagus increased with intraluminal pressure over the entire range of balloon volumes tested (50--300 microl). Distension-responsive neurons in the nucleus tractus solitarii showed rhythmic burst activity (type I), tonic excitation (type II), or inhibition followed by off bursts (type III). Increasing strength of stimulation changed type I responses to nonrhythmic but intensified type II and III responses. We conclude that load-dependent changes in distal esophagus motility pattern are encoded by vagal afferents alone and do not involve a spinal afferent input even at near-noxious stimulus strengths. PMID- 11408255 TI - Mechanism of folate transport across the human colonic basolateral membrane. AB - Previous studies from our laboratory have demonstrated the existence of a folate transporter in the human colonic apical membranes. The current studies were undertaken to examine the possible presence and function of a folate carrier in the human colonic basolateral membrane vesicles (BLMV). BLMV were purified from mucosal scrapings of colons of organ donors by a Percoll-density gradient centrifugation technique, and uptake studies were performed using a rapid filtration technique. Our results on [(3)H]Pte-Glu uptake are summarized as follows: 1) uptake was sensitive to osmolarity of the incubation medium; 2) Na(+) removal from the incubation medium did not affect folate uptake into BLMV; 3) uptake was significantly increased with decreasing incubation buffer pH from 8 to 4; 4) uptake demonstrated saturation kinetics with an apparent Michaelis constant of 9.6 +/- 0.48 microM and a maximal velocity of 8.10 +/- 0.36 pmol x mg protein( 1) x 10 s(-1); 5) uptake was markedly inhibited by the structural analog methotrexate (inhibitory constant = 8.28 +/- 1.0 microM); 6) uptake into BLMV demonstrated a trans-stimulation phenomenon; 7) anion exchange inhibitors DIDS and SITS significantly inhibited folate uptake; and 8) uptake was potential insensitive, as voltage clamping of vesicles or making them inside positive with K(+)/valinomycin failed to influence folate uptake. Western blot analysis using purified human colonic basolateral membrane preparations and specific polyclonal antibodies against the human reduced folate carrier (hRFC) has shown expression of the hRFC protein at this membrane domain. These data demonstrate the existence of a pH-dependent, DIDS-sensitive, electroneutral, carrier-mediated mechanism for folate transport across the human colonic basolateral membranes. PMID- 11408256 TI - Biochemical, histological, and inhibitor studies of membrane carbonic anhydrase in frog gastric acid secretion. AB - Gastric acid secretion is dependent on carbonic anhydrase (CA). To define the role of membrane-bound CA, we used biochemical, histochemical, and pharmacological approaches in the frog (Rana pipiens). CA activity and inhibition by membrane-permeant and -impermeant agents were studied in stomach homogenates and microsomal fractions. H(+) secretion in the histamine-stimulated isolated mucosa was measured before and after mucosal addition of a permeant CA inhibitor (methazolamide) and before and after mucosal or serosal addition of two impermeant CA inhibitors of differing molecular mass: a 3,500-kDa polymer linked to aminobenzolamide and p-fluorobenzyl-aminobenzolamide (molecular mass, 454 kDa). Total CA activity of frog gastric mucosa is 2,280 U/g, of which 10% is due to membrane-bound CA. Membrane-bound CA retains detectable activity below pH 4. Histochemically, there is membrane-associated CA in surface epithelial, oxynticopeptic, and capillary endothelial cells. Methazolamide reduced H(+) secretion by 100%, whereas the two impermeant inhibitors equally blocked secretion by 40% when applied to the mucosal side and by 55% when applied to the serosal side. The presence of membrane-bound CA in frog oxynticopeptic cells and its relative resistance to acid inactivation and inhibition by impermeant inhibitors demonstrate that it subserves acid secretion at both the apical and basolateral sides. PMID- 11408257 TI - Differential activation of intestinal gene promoters: functional interactions between GATA-5 and HNF-1 alpha. AB - The effects of GATA-4, -5, and -6, hepatocyte nuclear factor-1 alpha (HNF-1 alpha) and -beta, and Cdx-2 on the rat and human lactase-phlorizin hydrolase (LPH) and human sucrase-isomaltase (SI) promoters were studied using transient cotransfection assays in Caco-2 cells. GATA factors and HNF-1 alpha were strong activators of the LPH promoters, whereas HNF-1 alpha and Cdx-2 were strong activators of the SI promoter, although GATA factors were also necessary for maximal activation of the SI gene. Cotransfection of GATA-5 and HNF-1 alpha together resulted in a higher activation of all three promoters than the sum of the activation by either factor alone, demonstrating functional cooperativity. In the human LPH promoter, an intact HNF-1 binding site was required for functional synergy. This study is the first to demonstrate 1) differential activation of the LPH and SI promoters by multiple transcription factors cotransfected singly and in combination and 2) that GATA and HNF-1 transcription factors cooperatively activate intestinal gene promoters. Synergistic activation is a mechanism by which higher levels of tissue-specific expression might be attained by overlapping expression of specific transcription factors. PMID- 11408258 TI - Nifedipine modulation of biliary GSH and GSSG/ conjugate efflux in normal and regenerating rat liver. AB - Canalicular glutathione secretion provides the major driving force for bile acid independent bile flow (BAIF), although the pathways involved are not established. The hypothesis that GSH efflux proceeds by a route functionally distinct from the high-affinity, low-capacity, mrp2-mediated pathway was tested by using perfused rat liver and three choleretic compounds that modify biliary secretion of GSH (the dihydropyridine nifedipine and organic anion probenecid) or GSSG [sodium nitroprusside (SNP)]. Whereas nifedipine (30 microM) stimulated GSH secretion and blocked SNP-stimulated GSSG efflux and choleresis, SNP (1 mM) was ineffective against nifedipine-stimulated GSH efflux or BAIF, suggesting that most GSSG exits through a GSH-inhibitable path independent of high-affinity GSSG/glutathione conjugate transport. Three observations support this proposal. SNP, but not nifedipine, significantly inhibited bromosulfophthalein (BSP, 1 microM) excretion. Probenecid (1 mM) blocked resting or nifedipine-stimulated GSH secretion but only weakly inhibited BSP excretion. Glutathione, but not BSP, efflux capacity was reduced following partial hepatectomy. We suggest GSH efflux is mediated by a high-capacity organic anion pathway capable of GSSG transport when its high-affinity route is saturated. PMID- 11408259 TI - Relationship between ultrasonically detected phasic antral contractions and antral pressure. AB - The relationships between gastric wall motion and intraluminal pressure are believed to be major determinants of flows within and from the stomach. Gastric antral wall motion and intraluminal pressures were monitored in five healthy subjects by concurrent antropyloroduodenal manometry and transabdominal ultrasound for 60 min after subjects drank 500 ml of clear soup. We found that 99% of antral contractions detected by ultrasound were propagated aborally, and 68% of contractions became lumen occlusive at the site of the ultrasound marker. Of the 203 contractions detected by ultrasound, 53% were associated with pressure events in the manometric reference channel; 86% of contractions had corresponding pressure events detectable somewhere in the antrum. Contractions that occluded the lumen were more likely to be associated with a pressure event in the manometric reference channel (P < 0.01) and to be of greater amplitude (P < 0.01) than non-lumen-occlusive contractions. We conclude that heterogeneous pressure event patterns in the antrum occur despite a stereotyped pattern of contraction propagation seen on ultrasound. Lumen occlusion is more likely to be associated with higher peak antral pressure events. PMID- 11408260 TI - IL-12 gene transfer alters gut physiology and host immunity in nematode-infected mice. AB - Immune responses elicited by nematode parasite infections are characterized by T helper 2 (Th2) cell induction. The immunologic basis for changes in intestinal physiology accompanying nematode infection is poorly understood. This study examined whether worm expulsion and associated goblet cell hyperplasia and muscle contractility share a similar immune basis by shifting the response from Th2 to Th1 using interleukin-12 (IL-12) overexpression. We used a single administration of recombinant adenovirus vector expressing IL-12 (Ad5IL-12) in Trichinella spiralis-infected mice. Ad5IL-12 administered 1 day after infection prolonged worm survival and inhibited infection-induced muscle hypercontractility and goblet cell hyperplasia. This was correlated with upregulated interferon-gamma (IFN-gamma) expression and downregulated IL-13 expression in the muscularis externa layer. We also observed increased IFN-gamma production and decreased IL-4 and IL-13 production from in vitro stimulated spleen and mesenteric lymph node cells of infected Ad5IL-12-treated mice. These results indicate that transfer and overexpression of the IL-12 gene during Th2-based nematode infection shifts the immune response toward Th1 and delays worm expulsion. Moreover, the immune response shift abrogated the physiological responses to infection, attenuating both muscle hypercontractility and goblet cell hyperplasia. These findings strongly indicate that worm expulsion, muscle hypercontractility, and goblet cell hyperplasia share a common immunologic basis and may be causally linked. PMID- 11408261 TI - Increased in vitro activation of EGFR by membrane-bound TGF-alpha from gastric and colonic mucosa of aged rats. AB - Although aging is associated with increased epidermal growth factor receptor (EGFR) tyrosine kinase activity in Fischer 344 rat gastric and colonic mucosa, the regulatory mechanisms for the age-related rise in EGFR tyrosine kinase are poorly understood. Transmembrane transforming growth factor-alpha (TGF-alpha) may modulate EGFR function through an autocrine/juxtacrine mechanism. The present study aimed to determine the contribution of membrane-bound precursors of TGF alpha in enhancing EGFR activation in the gastric and colonic mucosa during aging. The extent of EGFR tyrosine phosphorylation, a measure of EGFR activation, was substantially higher (300--350%) in the gastric and colonic mucosa of 23- (aged) vs. 4-mo-old (young) Fischer 344 rats. This was accompanied by an increase (200--1,000%) in the relative concentration of 18- to 20-kDa membrane-bound precursor forms of TGF-alpha. The amount of TGF-alpha bound to EGFR was also higher (150-250%) in the gastric and colonic mucosa of aged vs. young rats. In vitro studies revealed that exposure of HCT 116 cells (a colon cancer cell line) to TGF-alpha from gastric and colonic mucosal membranes of aged rats caused a 200 -250% higher activation of EGFR and extracellular signal-related kinases (p42/44) compared with young rats. Our data suggest that the membrane-bound precursor form(s) of TGF-alpha may partly be responsible for enhancing EGFR activation in the gastric and colonic mucosa of aged rats, probably though an autocrine/juxtacrine mechanism(s). PMID- 11408262 TI - Adaptive vasodilatory response after octreotide treatment. AB - Despite the suppression of glucagon release, an adaptive response aimed at maintaining vasodilatation after octreotide treatment may exist in portal hypertension. The present study was undertaken to evaluate the possible interaction between endothelium and non-endothelium-derived vasodilators after 1 wk octreotide administration in cirrhotic rats. Rats were allocated to receive either vehicle or octreotide (30 or 100 microg/kg every 12 h subcutaneously). Hemodynamic values, plasma glucagon levels, endothelium-related vasodilatory activities, and aortic endothelial nitric oxide synthase (eNOS) expression were determined after treatment. Octreotide administration decreased plasma glucagon and increased serum 6-keto-PGF(1 alpha) and NOx levels without affecting the hemodynamic values. In cirrhotic rats receiving octreotide, there was a blunt response to either L-NAME or indomethacin administration alone, but this blunt pressor response disappeared after simultaneous administration of the two drugs. Additionally, an increased aortic eNOS expression was observed in cirrhotic rats receiving 1-wk octreotide. It is concluded that 1-wk octreotide treatment did not correct the hemodynamic derangement in cirrhotic rats. The enhanced endothelium related vasodilatory activity was noted after octreotide treatment that overcame the octreotide-induced hemodynamic effects in portal hypertension. PMID- 11408263 TI - Adaptation of stress-induced mucosal pathophysiology in rat colon involves opioid pathways. AB - Acute stress increases ion secretion and permeability of rat colonic epithelium. However, it is not known if stress-induced mucosal changes are subject to adaptation. Wistar-Kyoto rats were exposed to either continuous water-avoidance stress (CS) for 60 min or intermittent stress (IS) for three 20-min periods. Distal colonic segments were mounted in Ussing Chambers, and ion-transport [short circuit current (I(sc))] and permeability [conductance and flux of horseradish peroxidase (HRP)] parameters were measured. CS significantly increased I(sc), conductance, and HRP flux compared with control values. In contrast, in IS rats these variables were similar to those in nonstressed controls. To study the pathways involved in IS-induced adaptation, rats were pretreated intraperitoneally with the opioid antagonists naloxone or methylnaloxone. Opioid antagonists had no effect on values in control or CS rats. However, in the IS group, naloxone and methylnaloxone reversed the adaptive responses, and all variables increased to CS values. We conclude that stress-induced colonic mucosal pathophysiology is subject to rapid adaptation, which involves opioid pathways. PMID- 11408264 TI - Insulin and IGF-I inhibit calcium-dependent chloride secretion by T84 human colonic epithelial cells. AB - D-Myo-inositol (3,4,5,6) tetrakisphosphate [Ins(3,4,5,6)P(4)] or phosphatidylinositol 3-kinase (PI 3-kinase) activity acts to inhibit calcium dependent chloride secretion in T84 colonic epithelial cells. To further distinguish between the contributions of these two signaling pathways to the inhibition of secretion, we studied effects of insulin, because the insulin receptor links to PI 3-kinase but not to pathways postulated to generate Ins(3,4,5,6)P(4). Chloride secretion across T84 cell monolayers was studied in Ussing chambers. Activation of PI 3-kinase was assessed by Western blotting. Basolateral, but not apical, addition of insulin inhibited carbachol- and thapsigargin-induced chloride secretion in a time- and concentration-dependent fashion. Insulin-like growth factor-I (IGF-I) had similar effects. Insulin had no effect on Ins(3,4,5,6)P(4) levels, and the inhibitory effects of insulin and IGF I on chloride secretion were fully reversed by the PI 3-kinase inhibitors wortmannin and LY-294002. Western blot analysis showed that both insulin and IGF I recruited the 85-kDa regulatory and 110-kDa catalytic subunits of PI 3-kinase to anti-phosphotyrosine immunoprecipitates. In conclusion, insulin and IGF-I act to inhibit calcium-dependent chloride secretion through a PI 3-kinase-dependent pathway. Because insulin is released in a pulsatile fashion postprandially and IGF-I levels are elevated in pathological settings, our findings may have physiological and/or pathophysiological significance. PMID- 11408265 TI - Mechanisms of intestinal gas retention in humans: impaired propulsion versus obstructed evacuation. AB - To explore the clinical role of intestinal gas dynamics, we investigated two potential mechanisms of gas retention, defective propulsion and obstructed evacuation. In healthy subjects, a gas mixture was continuously infused into the jejunum (4 ml/min) 1) during a 2-h control period of spontaneous gas evacuation and 2) during a 2-h test period either with impaired gut propulsion caused by intravenous glucagon (n = 6) or with obstructed (self-restrained) anal evacuation (n = 10) while anal gas evacuation, symptom perception (0-6 scale), and abdominal girth were measured. Impaired gut propulsion and obstructed evacuation produced similar gas retention (558 +/- 68 ml and 407 +/- 85 ml, respectively, vs. 96 +/- 58 ml control; P < 0.05 for both) and abdominal distension (8 +/- 3 mm and 6 +/- 3 mm, respectively, vs. 1 +/- 1 mm control; P < 0.05 for both). However, obstructed evacuation increased symptom perception (2.3 +/- 0.6 score change; P < 0.05), whereas gas retention in the glucagon-induced hypotonic gut was virtually unperceived (-0.4 +/- 0.7 score change; not significant). In conclusion, the perception of intestinal gas accumulation depends on the mechanism of retention. PMID- 11408266 TI - Mechanism of thiamine uptake by human colonocytes: studies with cultured colonic epithelial cell line NCM460. AB - Thiamine (vitamin B(1)) is essential for normal cellular functions and growth. Mammals cannot synthesize thiamine and thus must obtain the vitamin via intestinal absorption. The intestine is exposed to a dietary thiamine source and a bacterial source in which the vitamin is synthesized by the normal microflora of the large intestine. Very little is known about thiamine uptake in the large intestine. The aim of this study was, therefore, to address this issue. Our results with human-derived colonic epithelial NCM460 cells as a model system showed thiamine uptake to be 1) temperature- and energy dependent, 2) Na(+) independent, 3) increased with increasing buffer pH from 5 to 8 and after cell acidification but inhibited by amiloride, 4) saturable as a function of concentration, 5) inhibited by thiamine structural analogs but not by unrelated organic cations, and 6) inhibited by modulators of a Ca(2+)/calmodulin-mediated pathway. NCM460 cells and native human colonic mucosa expressed the recently cloned human thiamine transporter THTR-1 (product of the SLC19A2 gene) at both mRNA and protein levels. These results demonstrate for the first time that human NCM460 colonocytes possess a specific carrier-mediated system for thiamine uptake that appears to be under the regulation of an intracellular Ca(2+)/calmodulin mediated pathway. It is suggested that bacterially synthesized thiamine in the large intestine may contribute to thiamine nutrition of the host, especially toward cellular nutrition of the local colonocytes. PMID- 11408267 TI - Neural change in Trichinella-infected mice is MHC II independent and involves M CSF-derived macrophages. AB - Intestinal inflammation due to nematode infection impairs enteric cholinergic nerve function and induces hypercontractility of intestinal muscle. Macrophages have been implicated in the neural changes, but the subpopulation and mechanism involved are unknown. We examined whether macrophages alter nerves by virtue of their ability to activate lymphocytes via major histocompatibility complex (MHC) II-restricted antigen presentation. We also attempted to evaluate the role of macrophage subsets using op/op mice deficient in macrophage colony-stimulating factor (M-CSF). ACh release from the myenteric plexus was measured in MHC II- and M-CSF-deficient (op/op) mice infected with Trichinella spiralis. F4/80-positive macrophages and interleukin-1 beta were constitutively present in op/op and op/? mice but increased only in op/? mice postinfection. After infection, a marked suppression of ACh release occurred only in infected MHC II-deficient and op/? mice. Muscle hypercontractility remained evident in infected op/? mice. Treatment with M-CSF restored macrophage number, and this was accompanied by suppression of cholinergic nerve function during infection. Thus M-CSF plays a critical role in this model by recruiting a subset of macrophages that selectively suppresses enteric neural function. PMID- 11408268 TI - Human duodenal mucosal brush border Na(+)/H(+) exchangers NHE2 and NHE3 alter net bicarbonate movement. AB - The proximal duodenal mucosa secretes HCO that serves to protect the epithelium from injury. In isolated human duodenal enterocytes in vitro, multiple luminal membrane proteins are involved in acid/base transport. We postulated that one or more isoforms of the Na(+)/H(+) exchanger (NHE) family is located on the apical surface of human duodenal mucosal epithelial cells and thereby contributes to duodenal mucosal HCO transport. Duodenal biopsies were obtained from human volunteers, and the presence of NHE2 and NHE3 was determined by using previously characterized polyclonal antibodies (Ab 597 for NHE2 and Ab 1381 for NHE3). In addition, proximal duodenal mucosal HCO(3)(-) transport was measured in humans in vivo in response to luminal perfusion of graded doses of amiloride; 10(-5)--10( 4) M amiloride was used to inhibit NHE2 and 10(-3) M amiloride to inhibit NHE3. Both NHE2 and NHE3 were localized principally to the brush border of duodenal villus cells. Sequential doses of amiloride resulted in significant, step-wise increases in net duodenal HCO(3)(-) output. Inhibition of NHE2 with 10(-5) M and 10(-4) M amiloride significantly increased net HCO(3)(-) output. Moreover, there was an additional, equivalent increase (P < 0.05) in duodenal HCO(3)(-) output with 10(-3) M amiloride, which inhibited NHE3. We conclude that 1) NHE2 and NHE3 are localized principally to the brush border of human duodenal villus epithelial cells; 2) sequential inhibition of NHE2 and NHE3 isoforms resulted in step-wise increases in net HCO(3)(-) output; 3) NHE2 and NHE3 participate in human duodenal villus cell HCO(3)(-) transport; and 4) the contribution of NHE-related transport events should be considered when studying duodenal HCO(3)(-) transport processes. PMID- 11408269 TI - Effects of substance P on identified neurons of the rat dorsal motor nucleus of the vagus. AB - Previous evidence suggests that substance P (SP) activates subpopulations of neurons within the dorsal motor nucleus of the vagus (DMV). In this study we aimed at identifying these subpopulations in relation to their gastrointestinal projection organs or vagal branches and characterizing pharmacologically the SP response. Using whole cell patch-clamp recordings from identified gastrointestinal-projecting vagal motoneurons, we found that SP induced an inward current in all neuronal groups except for cecum-projecting cells. The lowest percentage of SP-responding neurons was found in fundus-projecting cells, where SP also had a concentration-response curve that was shifted to the left (P < 0.05). Independently from the projections, the SP response was reduced by sendide and MEN 10,376 and mimicked by a combination of [Sar(9)-Met(O(2))(11)]SP and alpha-neurokinin. SP and alpha-neurokinin also increased the frequency, but not the amplitude, of postsynaptic currents. In conclusion, we demonstrated that SP induces both pre- and postsynaptic effects on DMV neurons via activation of neurokinin NK(1) and NK(2) receptors. The magnitude of the SP response was correlated to the peripheral target organ. PMID- 11408270 TI - Scavenging nitric oxide reduces hepatocellular injury after endotoxin challenge. AB - Sustained upregulation of inducible nitric oxide (NO) synthase in the liver after endotoxin [lipopolysaccharide (LPS)] challenge may result in hepatocellular injury. We hypothesized that administration of a NO scavenger, NOX, may attenuate LPS-induced hepatocellular injury. Sprague-Dawley rats received NOX or saline via subcutaneous osmotic pumps, followed 18 h later by LPS challenge. Hepatocellular injury was assessed using biochemical assays, light, and transmission electron microscopy (TEM). Interleukin (IL)-6 mRNA was measured by RT-PCR. Tumor necrosis factor (TNF)-alpha protein expression was determined by immunohistochemistry. NOX significantly reduced serum levels of ornithine carbamoyltransferase and aspartate aminotransferase. TNF-alpha and IL-6 expression were increased in the livers of saline-treated but not NOX-treated rats. Although there was no difference between groups by light microscopy, TEM revealed obliteration of the space of Disse in saline-treated but not in NOX-treated animals. Electron paramagnetic resonance showed the characteristic mononitrosyl complex in NOX treated rats. We conclude that NOX reduces hepatocellular injury after endotoxemia. NOX may be useful in the management of hepatic dysfunction secondary to sepsis or other diseases associated with excessive NO production. PMID- 11408271 TI - Regression of cholangiocyte proliferation after cessation of ANIT feeding is coupled with increased apoptosis. AB - Cholangiocyte proliferation and loss through apoptosis occur in cholestatic liver diseases. Our aim was to determine the mechanisms of apoptosis in an animal model of ductal hyperplasia. Rats were fed alpha-naphthylisothiocyanate (ANIT) for 2 wk and subsequently fed normal chow for 1, 2, and 4 wk. Proliferation was assessed in sections by morphometry and in small and large cholangiocytes by proliferating cellular nuclear antigen immunoblots and measurement of cAMP levels. Apoptosis and reactive oxygen species (ROS) levels were also assessed. ANIT feeding increased small and large cholangiocyte proliferation and apoptosis. Cessation of ANIT feeding was associated with decreased proliferation and a further increase in apoptosis in small and large cholangiocytes. Cholangiocytes from ANIT-fed rats or exposed to ANIT in vitro showed increased apoptosis and ROS generation. ANIT induced duct injury results in enhanced proliferation and apoptosis in small and large cholangiocytes. The mechanism of ANIT-induced apoptosis may be due to ROS generation induced directly by ANIT. Our model has implications for understanding the pathophysiology of cholangiopathies (characterized by the coexistence of cholangiocyte apoptosis and proliferation). PMID- 11408272 TI - Epidermal growth factor inhibits glycylsarcosine transport and hPepT1 expression in a human intestinal cell line. AB - The human intestinal cell line Caco-2 was used as a model system to study the effects of epidermal growth factor (EGF) on peptide transport. EGF decreased apical-to-basolateral fluxes of [(14)C]glycylsarcosine ([(14)C]Gly-Sar) up to 50.2 +/- 3.6% (n = 6) of control values. Kinetic analysis of the fluxes showed that maximal flux (V(max)) of transepithelial transport decreased from 3.00 +/- 0.17 nmol x cm(-2) x min(-1) in control cells to 0.50 +/- 0.07 nmol x cm(-2) x min(-1) in cells treated with 5 ng/ml EGF (n = 6, P < 0.01). The apparent Michaelis-Menten constant (K(m)) was 2.71 +/- 0.31 mM (n = 6) in control cells and 1.89 +/- 0.28 mM (n = 6, not significantly different from control) in EGF treated cells. Similarly, apical uptake of [(14)C]Gly-Sar decreased in cells treated with EGF, with an ED(50) value of 0.36 +/- 0.06 ng/ml (n = 6) EGF and a maximal inhibition of 80 +/- 0.02% (n = 6). V(max) decreased from 2.61 +/- 0.4 to 1.06 +/- 0.1 nmol x cm(-2) x min(-1) (n = 3, P < 0.05), whereas K(m) remained constant. Basolateral Gly-Sar uptake showed no changes in V(max) or K(m) after EGF treatment (n = 3). RT-PCR showed a decrease in hPepT1 mRNA (using glucose-6 phosphate dehydrogenase mRNA as control) in cells treated with EGF. Western blotting indicated a decrease in hPepT1 protein in cell lysates. We conclude that EGF treatment decreases Gly-Sar transport in Caco-2 cells by decreasing the number of peptide transporter molecules in the apical membrane. PMID- 11408273 TI - Attenuation of CCl(4)-induced hepatic fibrosis by GdCl(3) treatment or dietary glycine. AB - The role of Kupffer cells in CCl(4)-induced fibrosis was investigated in vivo. Male Wistar rats were treated with phenobarbital and CCl(4) for 9 wk, and a group of rats were injected with the Kupffer cell toxicant gadolinium chloride (GdCl(3)) or were fed glycine, which inactivates Kupffer cells. After CCl(4) alone, the fibrosis score was 3.0 +/- 0.1 and collagen protein and mRNA expression were elevated, but GdCl(3) or glycine blunted these parameters. Glycine did not alter cytochrome P-450 2E1, making it unlikely that glycine affects CCl(4) metabolism. Treatment with GdCl(3) or glycine prevented CCl(4) induced increases in transforming growth factor (TGF)-beta 1 protein levels and expression. CCl(4) treatment increased alpha-smooth muscle actin staining (score 3.0 +/- 0.2), whereas treatment with GdCl(3) and glycine during CCl(4) exposure blocked this effect (1.2 +/- 0.5); there was no staining with glycine treatment. These results support previous in vitro data and demonstrate that treatment of rats with the selective Kupffer cell toxicant GdCl(3) prevents stellate cell activation and the development of fibrosis. PMID- 11408274 TI - Functional overlap of IP(3)- and cADP-ribose-sensitive calcium stores in guinea pig myenteric neurons. AB - In myenteric neurons two different receptor subtypes govern the intracellular Ca(2+) stores: the inositol 1,4,5-trisphosphate (IP(3)) receptor (IP(3)R) and the ryanodine receptor (RyR). Their degree of functional overlap was determined by examining Ca(2+) release in these cells through both superfusion techniques and intracellular microinjection. Microinjection of IP(3) (50 microM) and cADP-ribose (cADPr, 50 microM), specific ligands for the IP(3)R and RyR, respectively, demonstrated mobilization of intracellular Ca(2+) stores. Perfusion with cinnarizine (50 microM) or dantrolene (10 microM), antagonists of the IP(3)R and RyR, respectively, eliminated the Ca(2+) response to microinjected IP(3) and cADPr. Superfusion of the neurons with 100 microM ATP, an IP(3)-mediated Ca(2+) mobilizing agonist, caused intracellular Ca(2+) increments, which were antagonized by cinnarizine, and the RyR antagonists dantrolene, procaine (5 mM), and ryanodine (1 microM). Caffeine (10 mM) was applied repetitively in Ca(2+) free conditions to deplete RyR-sensitive stores; subsequent perfusion with ATP demonstrated a Ca(2+) response. Conversely, caffeine caused a Ca(2+) response after repetitive ATP exposures. The internal Ca(2+) stores of myenteric neurons are governed by two receptor subtypes, IP(3)R and RyR, which share partial functional overlap. PMID- 11408275 TI - Inflammatory bowel disease is associated with changes of enterocytic junctions. AB - Changes of the intestinal mucosal barrier are considered to play a role in the pathogenesis of inflammatory bowel disease (IBD). Our experiments were designed to identify dysregulation of epithelial junctional molecules in the IBD intestinum and to address whether altered expression of these molecules is a primary event in IBD or a phenomenon secondary to the inflammatory process. Noninflamed and inactively and actively inflamed mucosal tissues from patients with ulcerative colitis or Crohn's disease as well as tissues from control subjects were analyzed for the expression of junctional molecules by different methods. Marked downregulation of junctional proteins and their respective mRNAs was observed in actively inflamed IBD tissues. In IBD tissues with inactive inflammation, only a few junctional molecules such as E-cadherin and alpha catenin were affected, whereas expression of desmosomal or tight junction associated proteins appeared almost unchanged. In noninflamed IBD tissues, junctional protein expression was not different from that seen in normal control subjects. In IBD, downregulation of junctional molecule expression is apparently associated with the inflammatory process and does not likely represent a primary phenomenon. PMID- 11408276 TI - Differentiation of immature enterocytes into enteroendocrine cells by Pdx1 overexpression. AB - The development of a variety of enteroendocrine cells of the gut is poorly understood. We tested whether immature intestinal stem cells were switched to multiple enteroendocrine hormone-producing cells by in vitro transfer of a homeobox gene. We transfected the pancreatic-duodenal homeobox 1 gene (Pdx1) into IEC-6 cells, an embryonic intestinal epithelial cell line derived from a normal rat, and selected the cells that overexpressed Pdx1 by 150-fold compared with control. The cells were examined for differentiation into enteroendocrine cells by immunocytochemical and electron microscopic analyses. Transfected cells cultured on micropore filters formed a trabecular network piled up on monolayer cells. These trabecular cells showed nuclear localization of Pdx1 protein and contained well-developed rough endoplasmic reticulum as well as many secretory granules of pleomorphic shape in the cytoplasm. Antibodies against chromogranin A, serotonin, cholecystokinin, gastrin, and somatostatin stained these secretory granules in the cytoplasm. Furthermore, immunofluorescence double staining analysis showed that different hormones were produced within a cell. These results provide the evidence that immature intestinal epithelial cells can differentiate into multiple hormone-producing enteroendocrine cells in response to overexpression of Pdx1. PMID- 11408277 TI - Pre- and postsynaptic inhibition by nociceptin in guinea pig small intestinal myenteric plexus in vitro. AB - Actions of nociceptin on electrical and synaptic behavior of morphologically and neurochemically identified neurons in the guinea pig duodenal myenteric plexus were studied with conventional techniques. Nociceptin hyperpolarized the membrane potential in 104 of 121 AH-type and 28 of 51 S-type neurons with an EC(50) of 11.9 +/- 1.2 nM. Increased K(+) conductance accounted for the hyperpolarizing responses that were blocked by pertussis toxin and unaffected by naloxone. The selective opioid receptor-like (ORL)(1) receptor antagonist [Phe(1)-psi(CH(2)-NH) Gly(2)]nociceptin(1--13)-NH(2) suppressed the nociceptin-evoked responses while behaving like a partial agonist. The nonselective ORL(1) antagonist naloxone benzoylhydrazone competitively suppressed nociceptin actions with a pA(2) value of 5.8. Nociceptin acted at presynaptic inhibitory receptors to suppress fast excitatory nicotinic postsynaptic potentials in 25 of 30 neurons (EC(50) = 22.5 +/- 4.4 nM) and slow synaptic excitation in 38 of 45 neurons (EC(50) = 15.1 +/- 1.6 nM). Presynaptic inhibitory action of nociceptin was unaffected by naloxone and was antagonized by [Phe(1)-psi(CH(2)-NH)-Gly(2)]nociceptin(1--13)-NH(2) or naloxone benzoylhydrazone. The results suggest that nociceptin acts both pre- and postsynaptically by activating an ORL(1) receptor that is distinct from typical naloxone-sensitive opioid receptors. PMID- 11408278 TI - Identification and localization of aquaporin water channels in human salivary glands. AB - Aquaporin (AQP) water channels are expressed in a variety of fluid-transporting epithelia and are likely to play a significant role in salivary secretion. Our aim was to identify and localize the aquaporins expressed in human salivary glands. Total RNA was extracted from human parotid, submandibular, sublingual, and labial glands and from human brain. Expression of aquaporin mRNA was assessed by RT-PCR using specific primers for human AQP1, AQP3, AQP4, and AQP5. All four aquaporins were detected by RT-PCR in all of the glands, and the sequences were confirmed after further amplification with nested primers. Cleaned PCR products were then used as (32)P-labeled cDNA probes in a semiquantitative Northern blot analysis using glyceraldehyde-3-phosphate dehydrogenase as reference. Only AQP1, AQP3, and AQP5 mRNAs were present at significant levels. AQP localization was determined by immunohistochemistry on paraffin sections using affinity-purified primary antibodies and peroxidase-linked secondary antibodies. Each salivary gland type showed a broadly similar staining pattern: AQP1 was localized to the capillary endothelium and myoepithelial cells; AQP3 was present in the basolateral membranes of both mucous and serous acinar cells; AQP4 was not detected; and AQP5 was expressed in the luminal and canalicular membranes of both types of acinar cell. We conclude that AQP3 and AQP5 together may provide a pathway for transcellular osmotic water flow in the formation of the primary saliva. PMID- 11408279 TI - Evidence supporting presence of two pacemakers in rat colon. AB - Intracellular microelectrodes and organ bath techniques were used to study spontaneous cyclic electrical and mechanical activity in the rat colon. Electron microscopy and immunohistochemical studies showed two major populations of interstitial cells of Cajal (ICC): one associated with Auerbach's plexus (ICC-AP) and one with the submuscular plexus (ICC-SMP). The ICC-SMP network partly adhered to the submucosa when removed and was generally strongly damaged after separation of musculature and submucosa. Similarly, longitudinal muscle removal severely damaged AP. Two electrical and mechanical activity patterns were recorded: pattern A, low-frequency (0.5--1.5 cycles/min), high-amplitude oscillations; and pattern B, high-frequency (13--15 cycles/min), low-amplitude oscillations. Pattern A was recorded in preparations with intact AP but absent in those without intact AP. Pattern B was recorded in preparations with intact SMP but was absent in those lacking SMP. With full-thickness strips, the superimposed patterns A and B were recorded in circular muscle. When longitudinal muscle mechanical activity was recorded, only pattern A was present. We conclude that two pacemakers regulate rat colonic cyclic activity: the ICC-SMP network (responsible for cyclic slow waves and small-amplitude contractions) and the ICC-AP network (which may drive the cyclic depolarizations responsible for high-amplitude contractions). This is the first report showing consistent slow wave activity in the rodent colon. PMID- 11408280 TI - Free and protein-bound glutamine have identical splanchnic extraction in healthy human volunteers. AB - The objectives of the present study were to determine the splanchnic extraction of glutamine after ingestion of glutamine-rich protein ((15)N-labeled oat proteins) and to compare it with that of free glutamine and to determine de novo glutamine synthesis before and after glutamine consumption. Eight healthy adults were infused intravenously in the postabsorptive state with L-[1-(13)C]glutamine (3 micromol x kg(-1) x h(-1)) and L-[1-(13)C]lysine (1.5 micromol x kg(-1) x h( 1)) for 8 h. Four hours after the beginning of the infusion, subjects consumed (every 20 min) a liquid formula providing either 2.5 g of protein from (15)N labeled oat proteins or a mixture of free amino acids that mimicked the oat-amino acid profile and contained L-[2,5-(15)N(2)]glutamine and L-[2-(15)N]lysine. Splanchnic extraction of glutamine reached 62.5 +/- 5.0% and 66.7 +/- 3.9% after administration of (15)N-labeled oat proteins and the mixture of free amino acids, respectively. Lysine splanchnic extraction was also not different (40.9 +/- 11.9% and 34.9 +/- 10.6% for (15)N-labeled oat proteins and free amino acids, respectively). The main conclusion of the present study is that glutamine is equally bioavailable when given enterally as a free amino acid and when protein bound. Therefore, and taking into consideration the drawbacks of free glutamine supplementation of ready-to-use formulas for enteral nutrition, protein sources naturally rich in this amino acid are the best option for providing stable glutamine. PMID- 11408281 TI - Neural regulation of in vitro giant contractions in the rat colon. AB - The rat middle colon spontaneously generates regularly occurring giant contractions (GCs) in vitro. We investigated the neurohumoral and intracellular regulation of these contractions in a standard muscle bath. cGMP content was measured in strips and single smooth muscle cells. The circular muscle strips generated spontaneous GCs. Their amplitude and frequency were significantly increased by tetrodotoxin (TTX), omega-conotoxin, N(omega)-nitro-L-arginine (L NNA), and the dopamine D(1) receptor antagonist Sch-23390. The GCs were unaffected by hexamethonium, atropine, and antagonists of serotonergic (5-HT(1- 4)), histaminergic (H(1--2)), and tachykininergic (NK(1--2)) receptors but enhanced by NK(3) receptor antagonism. The guanylate cyclase inhibitor 1H [1,2,4]oxidiazolo[4,3-a]quinoxalin-1-one (ODQ) also enhanced GCs to the same extent as TTX and L-NNA, and each of the three agents prevented the effects of the others. GCs were abolished by electrical field stimulation, S-nitroso-N acetyl-penicillamine, and 8-bromo-cGMP. BAY-K-8644 and apamin enhanced the GCs, but they were abolished by D-600. Basal cGMP content in strips was decreased by TTX, L-NNA, or ODQ, but these treatments had no effect on cGMP content of enzymatically dissociated single smooth muscle cells. We conclude that spontaneous contractions in the rat colonic muscle strips are not generated by cholinergic, serotonergic, or histaminergic input. Constitutive release of nitric oxide from enteric neurons sustains cGMP synthesis in the colonic smooth muscle to suppress spontaneous in vitro GCs. PMID- 11408282 TI - Vagal innervation modulates motor pattern but not initiation of canine gastric migrating motor complex. AB - To determine the role of vagal nerves in initiation and modulation of the gastric migrating motor complex (MMC), motor activity was recorded in four dogs before and after total abdominal vagotomy during fasting, after exogenous intravenous motilin and insulin, and after feeding. After vagotomy, a temporally coordinated cyclic gastric and small bowel MMC persisted with an unchanged period. During gastric phase III, vagotomy decreased number of contractions (42 +/- 4 vs. 16 +/- 2), number of groupings of contractions (14 +/- 1 vs. 7 +/- 1), and motility index (12 +/- 1 vs. 10 +/- 1) and increased the duration between groupings (1 +/- 1 vs. 3 +/- 1 min) (P < 0.05 in each). Before and after vagotomy, motilin and insulin induced a premature MMC with minor changes in contractile pattern. A 200 g liver meal but not a 50-g liver meal inhibited the gastric MMC after vagotomy. A cyclic MMC persisted after vagotomy, but the contractile pattern during gastric phase III was altered. After a short recovery period, vagal innervation to the stomach modulates the pattern but not the presence of gastric interdigestive motility during phase III. PMID- 11408283 TI - Overexpression of CYP27 in hepatic and extrahepatic cells: role in the regulation of cholesterol homeostasis. AB - In the liver, sterol 27-hydroxylase (CYP27) participates in the classic and alternative pathways of bile acid biosynthesis from cholesterol (Chol). In extrahepatic tissues, CYP27 converts intracellular Chol to 27-hydroxycholesterol (27OH-Chol), which may regulate the activity of 3-hydroxy-3-methylglutaryl CoA reductase (HMG-CoA-R). This study attempts to better define the role of CYP27 in the maintenance of Chol homeostasis in hepatic and extrahepatic cells by overexpressing CYP27 in Hep G2 cells and Chinese hamster ovary (CHO) cells through infection with a replication-defective recombinant adenovirus encoding for CMV-CYP27. After infection, CYP27 mRNA and protein levels increased dramatically. CYP27 specific activity also increased two- to fourfold in infected cells (P < or = 0.02), with a marked increase in conversion of [(14)C]Chol to [(14)C]27OH-Chol (approximately 150%; P < or = 0.01). Accumulation of 27OH-Chol in CHO cells was associated with a 50% decrease in HMG-CoA-R specific activity (P < or = 0.02). In infected Hep G2 cells, the significant increase in bile acid synthesis (46%; P < or = 0.006), which prevented the accumulation of intracellular 27OH-Chol, resulted in increased HMG-CoA-R activity (183%; P < or = 0.02). Overexpression of CYP27 in Hep G2 cells also increased acyl CoA cholesterol acyltransferase (71%, P < or = 0.02) and decreased cholesteryl ester hydrolase (55%, P < or = 0.02). In conclusion, CYP27 generates different physiological responses depending on cell type and presence or absence of bile acid biosynthetic pathways. PMID- 11408284 TI - Health and human rights. The BMA's latest handbook on human rights challenges us all. PMID- 11408285 TI - Electronic fetal monitoring. Is not necessary for low risk labours. PMID- 11408286 TI - Preventing renal failure in the critically ill. There are no magic bullets-just high quality intensive care. PMID- 11408287 TI - Glucosamine for osteoarthritis: magic, hype, or confusion? It's probably safe-but there's no good evidence that it works. PMID- 11408288 TI - Twenty years of AIDS, and no end in sight. A BMJ theme issue will refocus attention on this catastrophic epidemic. PMID- 11408289 TI - South Africa will not pay for antiretroviral therapy. PMID- 11408290 TI - GMC extends restrictions on Bristol heart surgeon. PMID- 11408294 TI - Doctor wins seat in fight to save hospital. PMID- 11408295 TI - Fake prescription drugs are flooding the United States. PMID- 11408296 TI - Tobacco industry to pay damages to smoker and insurance firm. PMID- 11408299 TI - Midlife vascular risk factors and Alzheimer's disease in later life: longitudinal, population based study. AB - OBJECTIVE: To examine the relation of midlife raised blood pressure and serum cholesterol concentrations to Alzheimer's disease in later life. DESIGN: Prospective, population based study. SETTING: Populations of Kuopio and Joensuu, eastern Finland. PARTICIPANTS: Participants were derived from random, population based samples previously studied in a survey carried out in 1972, 1977, 1982, or 1987. After an average of 21 years' follow up, a total of 1449 (73%) participants aged 65-79 took part in the re-examination in 1998. MAIN OUTCOME MEASURES: Midlife blood pressure and cholesterol concentrations and development of Alzheimer's disease in later life. RESULTS: People with raised systolic blood pressure (>/=160 mm Hg) or high serum cholesterol concentration (>/=6.5 mmol/l) in midlife had a significantly higher risk of Alzheimer's disease in later life, even after adjustment for age, body mass index, education, vascular events, smoking status, and alcohol consumption, than those with normal systolic blood pressure (odds ratio 2.3, 95% confidence interval 1.0 to 5.5) or serum cholesterol (odds ratio 2.1, 1.0 to 4.4). Participants with both of these risk factors in midlife had a significantly higher risk of developing Alzheimer's disease than those with either of the risk factors alone (odds ratio 3.5, 1.6 to 7.9). Diastolic blood pressure in midlife had no significant effect on the risk of Alzheimer's disease. CONCLUSION: Raised systolic blood pressure and high serum cholesterol concentration, and in particular the combination of these risks, in midlife increase the risk of Alzheimer's disease in later life. PMID- 11408300 TI - A cognitive behavioural intervention to reduce sexually transmitted infections among gay men: randomised trial. AB - OBJECTIVE: To determine the effectiveness of a brief cognitive behavioural intervention in reducing the incidence of sexually transmitted infections among gay men. DESIGN: Randomised controlled trial with 12 months' follow up. SETTING: Sexual health clinic in London. PARTICIPANTS: 343 gay men with an acute sexually transmitted infection or who reported having had unprotected anal intercourse in the past year. MAIN OUTCOME MEASURES: Number of new sexually transmitted infections diagnosed during follow up and self reported incidence of unprotected anal intercourse. RESULTS: 72% (361/499) of men invited to enter the study did so. 90% (308/343) of participants returned at least one follow up questionnaire or re-attended the clinic and requested a check up for sexually transmitted infections during follow up. At baseline, 37% (63/172) of the intervention group and 30% (50/166) of the control group reported having had unprotected anal intercourse in the past month. At 12 months, the proportions were 27% (31/114) and 32% ( 39/124) respectively (P=0.56). However, 31% (38/123) of the intervention group and 21% (35/168) of controls had had at least one new infection diagnosed at the clinic (adjusted odds ratio 1.66, 95% confidence interval 1.00 to 2.74). Considering only men who requested a check up for sexually transmitted infections, the proportion diagnosed with a new infection was 58% (53/91) for men in the intervention group and 43% (35/81) for men in the control group (adjusted odds ratio 1.84, 0.99 to 3.40). Using a regional database that includes information from 23 sexual health clinics in London, we determined that few participants had attended other sexual health clinics. CONCLUSIONS: This behavioural intervention was acceptable and feasible to deliver, but it did not reduce the risk of acquiring a new sexually transmitted infection among these gay men at high risk. Even carefully designed interventions should not be assumed to bring benefit. It is important to evaluate their effects in randomised trials with objective clinical end points. PMID- 11408301 TI - Randomised controlled trial of cardiotocography versus Doppler auscultation of fetal heart at admission in labour in low risk obstetric population. AB - OBJECTIVE: To compare the effect of admission cardiotocography and Doppler auscultation of the fetal heart on neonatal outcome and levels of obstetric intervention in a low risk obstetric population. DESIGN: Randomised controlled trial. SETTING: Obstetric unit of teaching hospital PARTICIPANTS: Pregnant women who had no obstetric complications that warranted continuous monitoring of fetal heart rate in labour. INTERVENTION: Women were randomised to receive either cardiotocography or Doppler auscultation of the fetal heart when they were admitted in spontaneous uncomplicated labour. MAIN OUTCOME MEASURES: The primary outcome measure was umbilical arterial metabolic acidosis. Secondary outcome measures included other measures of condition at birth and obstetric intervention. RESULTS: There were no significant differences in the incidence of metabolic acidosis or any other measure of neonatal outcome among women who remained at low risk when they were admitted in labour. However, compared with women who received Doppler auscultation, women who had admission cardiotocography were significantly more likely to have continuous fetal heart rate monitoring in labour (odds ratio 1.49, 95% confidence interval 1.26 to 1.76), augmentation of labour (1.26, 1.02 to 1.56), epidural analgesia (1.33, 1.10 to 1.61), and operative delivery (1.36, 1.12 to 1.65). CONCLUSIONS: Compared with Doppler auscultation of the fetal heart, admission cardiotocography does not benefit neonatal outcome in low risk women. Its use results in increased obstetric intervention, including operative delivery. PMID- 11408303 TI - Secondary prevention in 24, 431 patients with coronary heart disease: survey in primary care. PMID- 11408304 TI - Tackling organisational change in the new NHS. PMID- 11408305 TI - 10-minute consultation: Prostatic symptoms. PMID- 11408306 TI - Recent advances: Paediatrics. PMID- 11408308 TI - A family cluster of Chlamydia trachomatis infection. PMID- 11408309 TI - ABC of AIDS. HIV infection and AIDS in the developing world. PMID- 11408310 TI - Forest plots: trying to see the wood and the trees. PMID- 11408312 TI - Is there such a thing as a life not worth living? PMID- 11408322 TI - Paediatric head injury. PMID- 11408323 TI - Consciousness. AB - Consciousness is topical, for reasons including its renewed respectability among psychologists, rapid progress in the neuroscience of perception, memory and action, advances in artificial intelligence and dissatisfaction with the dualistic separation of mind and body. Consciousness is an ambiguous term. It can refer to (i) the waking state; (ii) experience; and (iii) the possession of any mental state. Self-consciousness is equally ambiguous, with senses including (i) proneness to embarrassment in social settings; (ii) the ability to detect our own sensations and recall our recent actions; (iii) self-recognition; (iv) the awareness of awareness; and (v) self-knowledge in the broadest sense. The understanding of states of consciousness has been transformed by the delineation of their electrical correlates, of structures in brainstem and diencephalon which regulate the sleep-wake cycle, and of these structures' cellular physiology and regional pharmacology. Clinical studies have defined pathologies of wakefulness: coma, the persistent vegetative state, the 'locked-in' syndrome, akinetic mutism and brain death. Interest in the neural basis of perceptual awareness has focused on vision. Increasingly detailed neuronal correlates of real and illusory visual experience are being defined. Experiments exploiting circumstances in which visual experience changes while external stimulation is held constant are tightening the experimental link between consciousness and its neural correlates. Work on unconscious neural processes provides a complementary approach. 'Unperceived' stimuli have detectable effects on neural events and subsequent action in a range of circumstances: blindsight provides the classical example. Other areas of cognitive neuroscience also promise experimental insights into consciousness, in particular the distinctions between implicit and explicit memory and deliberate and automatic action. Overarching scientific theories of consciousness include neurobiological accounts which specify anatomical or physiological mechanisms for awareness, theories focusing on the role played by conscious processes in information processing and theories envisaging the functions of consciousness in a social context. Whether scientific observation and theory will yield a complete account of consciousness remains a live issue. Physicalism, functionalism, property dualism and dual aspect theories attempt to do justice to three central, but controversial, intuitions about experience: that it is a robust phenomenon which calls for explanation, that it is intimately related to the activity of the brain and that it has an important influence on behaviour. PMID- 11408324 TI - Neuropathology of inflicted head injury in children. I. Patterns of brain damage. AB - Fifty-three cases of non-accidental head injury in children were subjected to detailed neuropathological study, which included immunocytochemistry for microscopic damage. Clinical details were available for all the cases. There were 37 infants, age at head injury ranging from 20 days to 9 months, and 16 children (range 13 months to 8 years). The most common injuries were skull fractures (36% of cases), acute subdural bleeding (72%) and retinal haemorrhages (71%); the most usual cause of death was raised intracranial pressure secondary to brain swelling (82%). On microscopy, severe hypoxic brain damage was present in 77% of cases. While vascular axonal damage was found in 21 out of 53 cases, diffuse traumatic axonal injury was present in only three. Eleven additional cases, all of them infants, showed evidence of localized axonal injury to the craniocervical junction or the cervical cord. When the data were analysed by median age at head injury, statistically significant patterns of age-related damage emerged. Our study shows that infants of 2-3 months typically present with a history of apnoea or other breathing abnormalities, show axonal damage at the craniocervical junction, and tend also to have a skull fracture, a thin film of subdural haemorrhage, but lack extracranial injury. Children over 1 year are more likely to suffer severe extracranial, particularly abdominal, injuries. They tend to have larger subdural haemorrhages, and where traumatic axonal injury is present, show patterns of hemispheric white matter damage more akin to those reported in adults. Diffuse axonal injury is an uncommon sequel of inflicted head injury in children. PMID- 11408325 TI - Neuropathology of inflicted head injury in children. II. Microscopic brain injury in infants. AB - There are very few reports in the literature dealing with the neuropathology of infant head injury, and the question of whether diffuse traumatic brain damage [diffuse axonal injury (DAI)] occurs in such children has not yet been reliably established by detailed neuropathological studies. We report the findings in the brains of a series of 37 infants aged 9 months or less, all of whom died from inflicted head injuries, and 14 control infants who died of other causes. Axonal damage was identified using immunohistochemistry for beta-amyloid precursor protein. Full clinical details were available for each case, the most constant of which in the study cohort was an episode of significant apnoea at presentation, found to have been recorded in 75% of cases. Global hypoxic damage was the most common histological finding. Widespread axonal damage, interpreted as vascular, was present in 13 cases, but widespread traumatic axonal injury was found in only two children, both of whom had severe head injuries with multiple skull fractures. Epidural cervical haemorrhage and focal axonal damage to the brainstem and the spinal nerve roots, found in 11 cases but not in controls, indicate that the craniocervical junction is vulnerable in infant head injury, the neuropathology being that of stretch injury from cervical hyperextension/flexion. Damage to this region could account for the observed apnoea, which could in turn lead to hypoxic damage and brain swelling. The observation that the predominant histological abnormality in cases of inflicted head injury in the very young is diffuse hypoxic brain damage, not DAI, can be explained in one of two ways: either the unmyelinated axon of the immature cerebral hemispheres is relatively resistant to traumatic damage, or in shaking-type injuries the brain is not exposed to the forces necessary to produce DAI. PMID- 11408326 TI - Functional plasticity of language-related brain areas after cochlear implantation. AB - Using PET, the cerebral network engaged by heard language processing in normal hearing subjects was compared with that in patients who received a cochlear implant after a period of profound deafness. The experimental conditions were words, syllables and environmental sounds, each controlled by a noise baseline. Four categories of effect were observed: (i) regions that were recruited by patients and controls under identical task conditions: the left and right superior temporal cortices and the left insula were activated in both groups in all conditions; (ii) new regions, which were recruited by patients only: the left dorsal occipital cortex showed systematic activation in all conditions versus noise baselines; (iii) regions that were recruited by both groups with a different functional specificity; e.g. Wernicke's area responded specifically to speech sounds in controls but was not specialized in patients; and (iv) regions that were activated in one group more than the other: the precuneus and parahippocampal gyrus (patients more than controls) and the left inferior frontal, left posterior inferior temporal and left and right temporoparietal junction regions (controls more than patients). These data provide evidence for altered functional specificity of the superior temporal cortex, flexible recruitment of brain regions located within and outside the classical language areas and automatic contribution of visual regions to sound recognition in implant patients. PMID- 11408327 TI - Development of the hippocampal formation from 2 to 42 years: MRI evidence of smaller area dentata in autism. AB - Autism, a neuropsychiatric disorder that severely impairs social, language and cognitive development, has a clinical onset in the first years of life. Because components of the limbic system mediate memory, social and affective functions that are typically disturbed in autism, a developmental defect in the limbic system has been hypothesized to underlie different autistic symptoms, but no developmental study has been performed. To obtain neuroanatomical evidence of limbic system abnormality in autism, we measured the cross-sectional area of the area dentata (AD; dentate gyrus + CA4) and combined area of the subiculum and CA1 CA3 (CAS) using in vivo MRI. Autistic patients aged 29 months to 42 years (n = 59) and healthy normal controls (n = 51) participated. The cross-sectional area of the AD was significantly smaller than normal in autism, the largest deviation from normal size (-13.5%) being found in autistic children aged 29 months to 4 years. Strong age-related increases were seen in the cross-sectional area of CAS, but autistic and normal subjects were not significantly different. This is the first direct evidence that anatomical abnormality within the limbic system exists from the earliest years of the disorder, and persists throughout development and to middle age. PMID- 11408328 TI - Intrathecal antibody production against Chlamydia pneumoniae in multiple sclerosis is part of a polyspecific immune response. AB - Chronic intrathecal immunoglobulin (Ig) production is a hallmark of multiple sclerosis characterized by the presence of oligoclonal IgGs and, in addition, polyspecific recognition of different pathogens such as measles, rubella and herpes zoster virus. While the antigen specificity of the oligoclonal IgGs in multiple sclerosis is largely unknown, the oligoclonal IgGs arising during CNS infectious diseases are reactive against the specific pathogen. Recently, a link between Chlamydia pneumoniae and multiple sclerosis has been claimed. To test the possible role of C. pneumoniae in multiple sclerosis, we analysed (i) whether there is intrathecal IgG production against C. pneumoniae in multiple sclerosis and (ii) if the oligoclonal IgGs in the CSF of multiple sclerosis patients recognize C. pneumoniae. By studying paired serum-CSF samples from 120 subjects (definite multiple sclerosis, 46; probable multiple sclerosis, 12; other inflammatory neurological diseases, 35; other neurological diseases, 27) by enzyme-linked immunosorbent assay, we found that 24% of all patients with definite multiple sclerosis, but only 5% of patients with other inflammatory or non-inflammatory diseases, produced IgGs specific for C. pneumoniae intrathecally (definite multiple sclerosis versus other inflammatory neurological diseases: P = 0.027). The presence of intrathecal IgGs to C. pneumoniae was independent of the duration of disease and relatively stable over time. The major CSF oligoclonal IgG bands from multiple sclerosis patients with an intrathecal Ig production to C. pneumoniae did not react towards purified elementary bodies and reticulate bodies of C. pneumoniae on affinity-mediated immunoblot following isoelectric focusing (IEF-western blots). In contrast, the IgGs in the CSF of control patients with neuroborreliosis strongly reacted with their specific pathogen, Borrelia burgdorferi, by IEF-western blot analysis. Concomitant analysis of the CSF of 23 patients with a nested polymerase chain reaction for C. pneumoniae was negative in all cases. Together, our findings strongly suggest that the immune response to C. pneumoniae is part of a polyspecific intrathecal Ig production, as is commonly observed with other pathogens. This argues against a specific role for C. pneumoniae in multiple sclerosis. PMID- 11408329 TI - Effect of multiple subpial transection on motor cortical excitability in cortical dysgenesis. AB - We report here a 12-year-old patient with unilateral cortical dysgenesis and intractable simple partial seizure in his left arm, who underwent multiple subpial transection (MST) in the right cerebral cortex including the primary motor cortex. We investigated motor cortical excitability using multimodal transcranial magnetic stimulation (TMS) before and 1 month after MST, in which surgical cortical incisions were made with strokes 5 mm apart and 4 mm deep. Preoperative TMS studies showed hyperexcitability in the affected motor cortex as abnormally prolonged muscle responses to TMS with a wide cortical motor map, which were markedly reduced following the operation. The preoperative motor evoked potentials were large and polyphasic, and consisted of early and late components. The late component was completely abolished after MST, suggesting that this component might be due to activation of the corticospinal tract neurones by long recurrent axon branches of dysplastic excitatory pyramidal neurones, which were cut by MST, or by delayed, polysynaptic intracortical conduction with marked temporal dispersion. Intracortical inhibition in the affected motor cortex was also disrupted preoperatively and improved after MST. Postoperative recruitment order of muscle responses to TMS was bilaterally symmetrical, indicating that MST did not interfere with the function of the corticospinal tract neurones. The patient showed fair motor recovery and good seizure control after the operation. These results of TMS studies demonstrated the remarkable effectiveness of MST not only on intractable seizure but also on abnormal motor cortical organization and hyperexcitability in cortical dysgenesis. PMID- 11408330 TI - Persistent reelin-expressing Cajal-Retzius cells in polymicrogyria. AB - Cajal-Retzius (CR) cells are early-developing cells important in mammalian corticogenesis. Reelin, a protein secreted by CR cells, is essential for completion of neuronal migration and cortical lamination. Lack of reelin causes the 'reeler' phenotype in mice and autosomal recessive lissencephaly with cerebellar hypoplasia in man. Focal increases in reelin and CR cells are associated with thickening and local invaginations of the marginal zone and microgyria in animal studies. It has been suggested that abnormalities of reelin expression may be involved in human polymicrogyria. We have studied CR cells and reelin expression in pathological sections of human polymicrogyria to explore this possibility. Occurrence, distribution, morphology and reelin expression in CR cells were studied in 12 cases of human polymicrogyria, ranging from 21 gestational weeks to 10 years of age. Findings were compared with age-matched controls. Large, reelin-positive CR-like cells were more numerous in the majority of the polymicrogyria cases and persisted for longer than usual, up to 10 years of age. The CR-like cells tended to cluster and were most frequent in fused molecular layers in the polymicrogyria. Reelin-expressing CR-like cells were also found in bridges between the molecular layer and overlying leptomeningeal heterotopia and within the heterotopia itself. Clusters of CR-like cells were also found in adjacent non-polymicrogyric cortex. No clusters were seen in the control subjects. Increased numbers of CR-like cells were seen in both familial and acquired cases. In contrast to previous reports, the findings show that large CR-like cells persisted for longer than usual, up to 10 years of age, and that they may continue to express reelin. Their maximal aggregation in regions of polymicrogyria and overlying leptomeningeal heterotopia suggest an association between the presence of these cells and polymicrogyria, which we interpret in the light of recent findings concerning the roles of reelin and its downstream signalling pathway in neuronal and glial developmental dynamics and post developmental function. PMID- 11408331 TI - End-plate gamma- and epsilon-subunit mRNA levels in AChR deficiency syndrome due to epsilon-subunit null mutations. AB - Acetylcholine receptor (AChR) deficiency is the most common of the congenital myasthenic syndromes (CMS). Typically, the number of AChRs, measured by alpha bungarotoxin binding, is reduced to 10-30% of normal levels, the miniature end plate potentials are correspondingly reduced, and there are morphological changes at the motor end-plates. The majority of these syndromes are due to either missense or frameshift mutations within the gene encoding the adult-specific epsilon-subunit. These are often null mutations, but some mutant epsilon-subunits can be incorporated, at low levels, into functional AChRs in transfected cell lines. It is not clear, therefore, whether upregulation of the mutant epsilon subunit mRNA could generate sufficient AChR to support neuromuscular transmission, albeit at a reduced level. Conversely, it might be that the mutant epsilon-subunit transcripts are subject to mRNA surveillance and 'nonsense mediated' loss, leading to reduced epsilon-subunit mRNA expression. In either case, it is thought that neuromuscular transmission may be provided partly or entirely by incorporation of the foetal-specific gamma-subunit into end-plate AChR. gamma-Subunit mRNA is expressed at low levels in normal human muscle, but might be upregulated in CMS. The study of mRNA levels for AChR subunits should improve our understanding of genotype-phenotype relationships in CMS. Here we have defined homozygous epsilon-subunit mutations in four unrelated families with AChR deficiency and studied the steady-state levels of mRNA for AChR subunits at the motor end-plates by in situ hybridization. Although we demonstrated that each mutation would lead to almost complete absence of surface adult AChR expression, we detected similar robust expression of alpha- and epsilon-subunit mRNAs at end plates of patient and control muscles, suggesting that mRNA transcripts for the epsilon-subunit are neither upregulated nor degraded preferentially. Interestingly, we were unable to detect any increase in gamma-subunit mRNA expression at CMS end-plates. Transgenic mice lacking the epsilon-subunit die 2-3 months after birth, suggesting that alpha(2)betadelta(2) pentamers cannot sustain neuromuscular transmission. Therefore, we tentatively conclude that the persistent low level expression of the gamma-subunit, which is present in normal human muscles as well as in AChR deficiency syndromes, is sufficient to enable patients with epsilon-subunit null alleles to survive. PMID- 11408332 TI - Left minineglect in dyslexic adults. AB - We searched for a core mechanism underlying the diverse behavioural and sensorimotor deficits in dyslexic subjects. In psychophysical temporal order judgement and line motion illusion tasks, adult dyslexics processed stimuli in the left visual hemifield significantly (approximately 15 ms) more slowly than normal readers, indicating a left-sided 'minineglect'. Furthermore, abrupt stimuli captured attention in both visual hemifields less effectively in dyslexics than in normal readers. These abnormalities could reflect right parietal lobe hypofunction, a consequence of a general magnocellular deficit demonstrated previously. Based on these and previous data, we propose a causal chain which could result in several sensory and cognitive deficits observed in dyslexic subjects. PMID- 11408333 TI - Impaired temporal contrast sensitivity in dyslexics is specific to retain-and compare paradigms. AB - Developmental dyslexia is a specific reading disability that affects 5-10% of the population. Recent studies have suggested that dyslexics may experience a deficit in the visual magnocellular pathway. The most extensively studied prediction deriving from this hypothesis is impaired contrast sensitivity to transient, low luminance stimuli at low spatial frequencies. However, the findings are inconsistent across studies and even seemingly contradictory. In the present study, we administered several different paradigms for assessing temporal contrast sensitivity, and found both impaired and normal contrast sensitivity within the same group of dyslexic participants. Under sequential presentation, in a temporal forced choice paradigm, dyslexics showed impaired sensitivity to both drifting and flickering gratings. However, under simultaneous presentation, with a spatial forced choice paradigm, dyslexics' sensitivity did not differ from that of the controls. Within each paradigm, dyslexics' sensitivity was poorer at higher temporal frequencies, consistent with the magnocellular hypothesis. These results suggest that a basic perceptual impairment in dyslexics may be their limited ability to retain-and-compare perceptual traces across brief intervals. PMID- 11408334 TI - T(1) hypointense lesions in secondary progressive multiple sclerosis: effect of interferon beta-1b treatment. AB - Recently, the clinical efficacy of interferon beta-1b (IFNbeta-1b) was demonstrated for secondary progressive (SP) multiple sclerosis in a European multicentre study. We evaluated the effect of IFNbeta-1b treatment on the rate of development of hypointense T(1) MRI lesions, a putative marker of axonal damage. Unenhanced T(1)-weighted images were obtained in a subgroup of 95 multiple sclerosis patients from five centres at 6-month intervals; this subgroup was similar to the total study population for all demographic, clinical and MRI parameters. An experienced observer blinded to the clinical data and treatment allocation measured volumes. The median baseline lesion load for hypointense T(1) lesions was 5.1 cm(3) for placebo-treated and 4.9 cm(3) for IFNbeta-1b-treated patients (P = 0.56). Placebo-treated patients showed an increase in T(1) lesion load by a median of 14% per year (P = 0.0002 compared with baseline); this was reduced to 7.7% per year in the IFNbeta-1b-treated patients (P = 0.003 versus placebo). In the IFNbeta-1b arm there was a statistically significant correlation between absolute change in Expanded Disability Status Scale scores and T(1) lesion load by month 36 (r = 0.38, P = 0.0015). In patients with SP multiple sclerosis, IFNbeta-1b treatment reduces the development of hypointense T(1) lesions, suggesting that reduced axonal damage in lesions may play a part in the beneficial effect that is observed clinically. PMID- 11408335 TI - Spontaneous remyelination following extensive demyelination is associated with improved neurological function in a viral model of multiple sclerosis. AB - A major question in neurobiology is whether myelin repair can restore neurological function following the course of a severe, progressive CNS demyelinating disease that induces axonal loss. In this study we used Theiler's murine encephalomyelitis virus (TMEV) to induce a chronic progressive CNS demyelinating disease in mice that was immune-mediated and pathologically similar to human multiple sclerosis. Because immunosuppression of chronically TMEV infected mice has been shown to enhance myelin repair, we first addressed the potential roles of CD4(+) and CD8(+) T cells in the inhibition of CNS remyelination during chronic disease. TMEV infection of susceptible PL/J mice deficient in CD4(+) but not CD8(+) T cells demonstrated a significant increase in severity of pathogenesis when compared with wild-type controls. This was characterized by enhanced demyelination, spinal cord atrophy, neurological deficits, and mortality. Interestingly, the PL/J CD4(-/-) mice that survived to the chronic stage of the disease had nearly complete spontaneous myelin repair mediated by both oligodendrocytes and infiltrating Schwann cells. Therefore, we next addressed whether this spontaneous myelin repair was associated with improved neurological function despite the increased pathology. Of interest, all surviving PL/J CD4(-/-) mice showed partial restoration of motor coordination and gait that coincided temporally with spontaneous myelin repair. Furthermore, functional recovery of motor coordination correlated strongly with the percentage of myelin repair mediated by Schwann cells, whereas restoration of hindlimb gait correlated with oligodendrocyte-mediated myelin repair. This is the first study to demonstrate that spontaneous remyelination correlates with partial restoration of neurological function during the course of a progressive, immune-mediated CNS demyelinating disease. Of greater importance, functional recovery occurred despite previous severe demyelination and spinal cord atrophy. PMID- 11408336 TI - Contextual guidance of attention: human intracranial event-related potential evidence for feedback modulation in anatomically early temporally late stages of visual processing. AB - We investigated attentional guidance in early visual areas in the brain by recording event-related potentials directly from the surface of visual cortex. Patients performed a contextual cueing task in which attentive search to targets was guided by implicitly learned spatial context information. The earliest activity in striate cortex (area V1) was not modulated by contextual cueing, whereas later activity beginning at approximately 200 ms was enhanced by contextual cueing in V1, V2 and other portions of extrastriate cortex. These results suggest that context can enhance visual processing by temporally late top down modulation of activity in anatomically early areas of visual cortex. Together with anatomical and neurophysiological studies in animals, these results suggest an excitatory feedback mechanism acting on apical dendrites of pyramidal cells in V1 and other areas of visual cortex. PMID- 11408337 TI - Clinical, biochemical and molecular genetic characteristics of 19 patients with the Sjogren-Larsson syndrome. AB - Sjogren-Larsson syndrome (SLS) is an autosomal recessively inherited neurocutaneous disorder caused by a deficiency of the microsomal enzyme fatty aldehyde dehydrogenase (FALDH). We report the clinical characteristics and the results of molecular studies in 19 SLS patients. Patients 1-17 show the classical triad of severe clinical abnormalities including ichthyosis, mental retardation and spasticity. Most patients were born preterm, and all patients exhibit ocular abnormalities and pruritus. Electro-encephalography shows a slow background activity, without other abnormalities. MRI of the brain shows an arrest of myelination, periventricular signal abnormalities of white matter and mild ventricular enlargement. Cerebral (1)H-MR spectroscopy reveals a characteristic, abnormal lipid peak. The degree of white matter abnormality in the MRIs and the height of the lipid peak in (1)H-MR spectra do not correlate with the severity of the neurological signs. The clinical presentation and the clinical course is strikingly similar in these patients. Patient 18 shows a mild phenotype that essentially contains the same, but less severe, clinical features. Patient 19 exhibits the typical, but very mild, dermatological and ocular abnormalities, without any clinical neurological involvement. The diagnosis of SLS was confirmed by demonstration of the enzyme defect in cultured skin fibroblasts. Furthermore, as might be predicted from the essential role of FALDH in leucotriene B(4) (LTB(4)) metabolism, elevated urinary concentrations of LTB(4) and 20-OH-LTB(4) were found in all patients studied. Molecular studies of the FALDH gene revealed eight different mutations, including three new ones: a large 26-base pair deletion (21-46del), a missense mutation (80C-->T) and an insertion mutation (487 488insA). The vast majority of SLS patients seem to be severely affected independent of their genotype. PMID- 11408338 TI - The prevalence of progressive supranuclear palsy (Steele-Richardson-Olszewski syndrome) in the UK. AB - We performed a study to estimate the point prevalence of progressive supranuclear palsy (PSP) in the UK at national, regional and community levels. A 'Russian doll' design was used in which the population denominator for each of the three studies was successively smaller, whilst the method of case ascertainment became increasingly more rigorous. The NINDS-SPSP (National Institute of Neurological Disorders and the Society for Progressive Supranuclear Palsy) diagnostic criteria for PSP were applied throughout the study for case definition. The national study identified cases using passive referral mechanisms [e.g. the British Neurological Surveillance Unit (BNSU), PSP (Europe) Association patient register]. We identified 577 cases of PSP, giving a national prevalence estimate of 1.0 per 100 000 [95% confidence interval (CI) 0.9-1.1]. The North of England regional study used active 'multiple source' case ascertainment from a collaborative network of neurologists and non-neurologists. We identified 80 cases of PSP in this study, giving a crude and age-adjusted prevalence of 3.1 (95% CI 2.4-3.8) and 2.4 (1.9 3.0) per 100 000, respectively. Of these 80 cases, 51 patients (65%) were referred initially to non-neurologists and 10 patients (13%) had not seen a neurologist at any stage of their illness. The proportion of female cases was significantly greater in the regional than in the national study (61% versus 44%; P < 0.02). Cases referred to non-neurologists were significantly older than those referred to neurologists in the regional study (median age 73 versus 69.5 years; P < 0.01). Patients in the community study were identified via diagnostic and therapeutic registers from a representative sample of general practices in Newcastle upon Tyne. We identified 17 cases of PSP, yielding crude and age adjusted prevalences of 6.5 (95% CI 3.4-9.7) and 5.0 (95% CI 2.5-7.5) per 100 000, respectively. Seven of the 17 cases (41%) had not previously been diagnosed as PSP. This study suggests that PSP is more common than previously considered, is commonly misdiagnosed and that the majority of cases are not initially referred to neurologists. The study also confirms the importance of active and detailed case ascertainment in ensuring reliable prevalence estimates. PMID- 11408339 TI - Comprehension of metaphors and idioms in patients with Alzheimer's disease: a longitudinal study. AB - Language in patients with Alzheimer's disease has been extensively studied, with the exception of non-literal language comprehension. However, in our speech, we often make use of expressions, which are not necessarily interpreted on a literal ground. Comprehension of metaphors and idioms was examined in 39 patients with probable early Alzheimer's disease. The results showed that the decline of figurative language is not an early symptom of dementia and can occur independently from the impairment of propositional language. It was also found that metaphors and idioms differ as far as the predominant kind of error is concerned. PMID- 11408340 TI - Superoxide dismutase in CSF from amyotrophic lateral sclerosis patients with and without CuZn-superoxide dismutase mutations. AB - Mutations in CuZn-superoxide dismutase (CuZn-SOD) have been linked to familial amyotrophic lateral sclerosis (ALS), and motor neurone death is caused by the gain of a toxic property of the mutant protein. Here we determined amounts, activity and molecular forms of CuZn-SOD in CSF from ALS patients carrying the D90A and other CuZn-SOD mutations and patients without such mutations. There were no differences in amount of protein and enzymic activities of CuZn-SOD between 37 neurological controls, 54 sporadic and 12 familial ALS cases, and 10 cases homozygous for the D90A mutation. Three cases heterozygous for the A89V, S105L and G114A CuZn-SOD mutations showed low amounts of CuZn-SOD. There was no evidence for accumulation of inactive protein in any of the groups. Immunoblots showed no evidence for the presence of any precipitates or other molecular forms of CuZn-SOD with higher molecular weight in the groups. About 25% of the CuZn-SOD subunits in CSF from controls shows an N-terminal truncation. This truncated portion does not differ between controls and ALS groups not carrying CuZn-SOD mutations, but is 70% larger in samples from D90A homozygous ALS patients. The findings suggest an essentially normal amount and activity of D90A mutant CuZn SOD in CNS tissues of ALS cases. The increased occurrence of N-terminally truncated mutant subunits may indicate a difference in degradation routes compared with the wild-type enzyme, resistance against subsequent proteolytic steps and/or a compromised downstream proteolytic machinery. Molecular fragments accumulated to a greater extent from the D90A mutant enzyme might contribute to the motor neurone degeneration. We also determined the other SOD isoenzymes: in the controls, CuZn-SOD contributed 75%, extracellular SOD 25% and Mn-SOD <5% of the total SOD activity. There was no difference in the amount of extracellular SOD between any of the groups. PMID- 11408341 TI - Base excision repair in a network of defence and tolerance. PMID- 11408342 TI - Mechanisms of N-acetylcysteine in the prevention of DNA damage and cancer, with special reference to smoking-related end-points. AB - Although smoking cessation is the primary goal for the control of cancer and other smoking-related diseases, chemoprevention provides a complementary approach applicable to high risk individuals such as current smokers and ex-smokers. The thiol N-acetylcysteine (NAC) works per se in the extracellular environment, and is a precursor of intracellular cysteine and glutathione (GSH). Almost 40 years of experience in the prophylaxis and therapy of a variety of clinical conditions, mostly involving GSH depletion and alterations of the redox status, have established the safety of this drug, even at very high doses and for long-term treatments. A number of studies performed since 1984 have indicated that NAC has the potential to prevent cancer and other mutation-related diseases. N Acetylcysteine has an impressive array of mechanisms and protective effects towards DNA damage and carcinogenesis, which are related to its nucleophilicity, antioxidant activity, modulation of metabolism, effects in mitochondria, decrease of the biologically effective dose of carcinogens, modulation of DNA repair, inhibition of genotoxicity and cell transformation, modulation of gene expression and signal transduction pathways, regulation of cell survival and apoptosis, anti inflammatory activity, anti-angiogenetic activity, immunological effects, inhibition of progression to malignancy, influence on cell cycle progression, inhibition of pre-neoplastic and neoplastic lesions, inhibition of invasion and metastasis, and protection towards adverse effects of other chemopreventive agents or chemotherapeutical agents. These mechanisms are herein reviewed and commented on with special reference to smoking-related end-points, as evaluated in in vitro test systems, experimental animals and clinical trials. It is important that all protective effects of NAC were observed under a range of conditions produced by a variety of treatments or imbalances of homeostasis. However, our recent data show that, at least in mouse lung, under physiological conditions NAC does not alter per se the expression of multiple genes detected by cDNA array technology. On the whole, there is overwhelming evidence that NAC has the ability to modulate a variety of DNA damage- and cancer-related end-points. PMID- 11408343 TI - Myeloperoxidase--463A variant reduces benzo[a]pyrene diol epoxide DNA adducts in skin of coal tar treated patients. AB - The skin of atopic dermatitis patients provides an excellent model to study the role of inflammation in benzo[a]pyrene (BaP) activation, since these individuals are often topically treated with ointments containing high concentrations of BaP. In this study we have determined, by HPLC with fluorescence detection, the BaP diol epoxide (BPDE)-DNA adduct levels in human skin after topical treatment with coal tar and their modulation by the -463G-->A myeloperoxidase (MPO) polymorphism, which reduces MPO mRNA expression. BPDE-DNA adduct levels were 2.2 and 14.2 adducts/10(8) nt for MPO-463AA/AG and -463GG, respectively. The predominant BaP tetrol observed was tetrol I-1, which is derived after hydrolysis of the anti-BPDE-DNA adduct. The tetrol I-1/II-2 ratio, corresponding to the anti/syn ratio, was 6.7. The (32)P-post-labeling assay was also performed and thin layer chromatograms showed a major spot with a chromatographic location corresponding to BPDE-DNA. The mean values of the BPDE-DNA adduct spots were 3.8 +/- 2.4 per 10(8) nt for MPO-463AA/AG (n = 3) and 18.4 +/- 11.0 per 10(8) nt for MPO-463GG (n = 7), respectively (P = 0.03). One individual with the homozygous mutant genotype (-463AA) even had a 13-fold lower adduct level (1.4 per 10(8) nt) as compared to MPO-463GG subjects. In conclusion, these data show for the first time: (i) the in vivo formation of BPDE-DNA adducts in human skin treated with coal tar; (ii) that the MPO-463AA/AG genotype reduced BPDE-DNA adduct levels in human skin. PMID- 11408344 TI - Methylenetetrahydrofolate reductase C677T polymorphism does not alter folic acid deficiency-induced uracil incorporation into primary human lymphocyte DNA in vitro. AB - Methylenetetrahydrofolate reductase (MTHFR) is an enzyme which converts 5,10 methylene tetrahydrofolate (5,10-MnTHF) to 5-methyl tetrahydrofolate. A common C to T transition (C677T) in the MTHFR gene is reported to reduce the risk for colorectal cancer and acute lymphocytic leukemia in homozygotes (TTs). It is hypothesized that because TTs have reduced MTHFR activity, more 5,10-MnTHF is available to provide methyl groups for the conversion of uracil to thymidine. Folic acid deficiency causes the intracellular accumulation of dUMP and the subsequent incorporation of uracil into DNA. The removal of uracil from DNA may result in double-stranded DNA breaks, the accumulation of which is a putative risk factor for cancer. We tested whether human lymphocytes taken from TTs (n = 10) were more able to resist uracil incorporation into DNA than controls (n = 14 CCs and 6 CTs) when cultured in medium containing 12-120 nM folic acid for 9 days. DNA uracil content of these lymphocytes was measured by CG-MS. TTs and controls showed a dose-dependent increase in DNA uracil content during folic acid deficiency (P < 0.0001, R2 = 0.23 for TTs and P < 0.0001, R2 = 0.19 for controls). DNA uracil content was not different between the two groups at any of the folic acid concentrations (two-way ANOVA: media [folic acid], P < 0.0001; genotype, P = 0.4). The results show that, in this in vitro system, the MTHFR C677T polymorphism does not affect the cell's ability to resist uracil incorporation into DNA. Chromosome breakage, as measured by micronuclei, was also shown to correlate with folic acid concentration in a preliminary experiment (P < 0.0001). Although the results appear not to support the hypothesis that a reduced risk for certain cancers in TTs is due to diversion of folic acid to thymidine synthesis, differences between the in vivo and in vitro situation make this conclusion not definitive. PMID- 11408345 TI - Parous rats regain high susceptibility to chemically induced mammary cancer after treatment with various mammotropic hormones. AB - Parity in humans and rats provides significant protection against mammary tumor development. This study was carried out to investigate whether treatment of parous rats with mammotropic hormones would affect methyl-nitrosourea (MNU) induced mammary carcinogenesis. Parous rats were treated with 17beta-estradiol (E2), progesterone (P4) and thyroxine (T4) alone or in combination. E2 (20 microg/60 days) and P4 (20 mg/60 days) were administered by silastic tubing and T4 in the drinking water (3 microg T4/ml). Hormonal treatments commenced 7 days before MNU injection and continued for 33 weeks. Animals were palpated weekly for tumor detection. The effects of the hormonal treatments on the circulating concentrations of E2, P4, growth hormone (GH), prolactin (PRL), T4 and insulin like growth factor-I (IGF-I) after 7 days of treatment, the time of MNU injection, was assessed. Animals treated with E2 had significantly elevated circulation concentrations of GH, PRL and P4, and serum levels of E2 were more consistent in this group than in the other animal groups. P4 treatment caused elevation in P4 concentration in serum but did not affect the circulating levels of other hormones. The proliferation of the mammary gland at the time of MNU injection was elevated in animal groups treated with E2 either alone or with P4 and T4 and in animals treated with P4 alone, but the mammary gland was most differentiated in untreated parous rats and least in animals treated with E2 either alone or with P4 and T4. Mammary tumor incidence was 10% in parous rats that did not receive any hormonal treatment. Treatments with E2 or P4 alone significantly increased the susceptibility of parous animals to 67 and 50.0%, respectively; a tumor incidence similar to that of untreated AMV rats (64%). Parous rats treated with E2 plus P4 had tumor incidence higher than 90%. T4 administered did not affect mammary carcinogenesis. PMID- 11408346 TI - Hepatocyte growth factor induces colonic cancer cell invasiveness via enhanced motility and protease overproduction. Evidence for PI3 kinase and PKC involvement. AB - Tumour progression to the metastatic phenotype is mainly dependent on tumour cell invasiveness. Cell migration is a crucial step in this process. Here we investigate the effect of hepatocyte growth factor (HGF) on the induction of in vitro invasiveness of poorly aggressive Caco-2 colonic cancer epithelial cells. Invasion assays through a Matrigel barrier were performed. Proteases were assessed by zymography, reverse transcription-polymerase chain reaction and immunoblotting. Caco-2 cells were found to express HGF receptor but not HGF and to secrete several proteases, namely matrix metalloproteinase-1 (MMP-1), MMP-2, possibly MMP-9 and urokinase plasminogen activator (uPA). Exogenous HGF promoted invasiveness of Caco-2 cells through an artificial basement membrane matrix and enhanced their production of proteases. In addition, analyses of media at the end of invasion assays indicated that anti-HGF antibody inhibited protease production in parallel with cell invasion. The involvement of proteases in the HGF-induced invasion process was further investigated using either a synthetic general MMP inhibitor or neutralizing antibodies against MMPs or uPA. All components significantly inhibited HGF-promoted cell invasion. Moreover, specific inhibitors of PKCalpha/beta1 and PI3 kinase also decreased both HGF-promoted cell invasion and protease expression in invasion assay media. Thus, our findings provide evidence that the process of HGF-activated invasiveness of Caco-2 cells involves PI3 kinase and PKC and results from close association of two events, stimulation of cell motile activity and concomitant overproduction of proteases, which permits cell migration through a degraded extracellular matrix. PMID- 11408347 TI - Magnetic fields (MF) of 50 Hz at 1.2 microT as well as 100 microT cause uncoupling of inhibitory pathways of adenylyl cyclase mediated by melatonin 1a receptor in MF-sensitive MCF-7 cells. AB - Magnetic fields (MF) of 60 Hz at 1.2 microT were previously shown to inhibit the antiproliferative effect of melatonin on MCF-7 cells (Liburdy,R.P., 1993, J. Pimeal Res. 14, 89-97). In addition, three laboratories (Blackman,C.F. and Benane,S.G., 1998; Luben,R.A. and Morgan,A.P., 1998; Morris,J.E., Chrisler,W.B., Miller,D.L., Sasser,L.B. and Anderson,L.E., 1998; 20th Annual Meeting of the Bioelectromagnetics Society, At. Pete Beach, FL) independently reported results consistent with this finding. In this study, we investigated the molecular basis of the biological effects of MF using MCF-7 cells. Only 1a melatonin receptors were identified by the [125I]melatonin binding assay and RT-PCR analysis. Moreover, preceding exposures to MF of 100 microT for 3, 5 and 7 days blocked the melatonin-induced inhibition of cAMP accumulation in a time-dependent manner, while none of the melatonin receptor functions or GTPase and adenylyl cyclase activities were affected. Estrogen-evoked cell proliferation was not altered by MF either. Exposure to 1.2 microT MF exerted the same effects on the melatonin signaling pathway as that to 100 microT. Thus, this is the first study to provide evidence that MF may cause uncoupling of signal transduction from melatonin receptors to adenylyl cyclase. PMID- 11408348 TI - Association of matrilysin mRNA expression with K-ras mutations and progression in pancreatic ductal adenocarcinomas. AB - The matrix metalloproteinase matrilysin has been implicated in the progression of gastrointestinal and other cancers. The aim of this study was to examine matrilysin mRNA expression and determine whether it is correlated with K-ras mutations and/or progression of pancreatic carcinoma. Using the semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR), we analyzed 11 pancreatic cancer cell lines and 70 pancreatic adenocarcinoma tissues for matrilysin mRNA expression. The results were correlated with clinicopathological characteristics and K-ras mutations. Significant amounts of matrilysin mRNA were detected in six of the eight cell lines with K-ras mutations but not in the three cell lines with wild-type K-ras. Matrilysin mRNA was detected in 57 (81.4% ) of the 70 tumor tissues and in all of the eight liver metastases, but not in any of the adjacent non-tumorous tissues. Matrilysin expression was significantly correlated with the size of tumor, tumor spreading, lymph node metastasis, advanced pathologic tumor-node- metastasis stage and K-ras mutations. The relative amounts of matrilysin mRNA in tumor tissues increased with increase in tumor stage and were highest in liver metastatic tumor tissues. Our results suggest that matrilysin, the expression of which is correlated with K-ras mutations, plays a key role in tumor growth and progression of pancreatic carcinoma. PMID- 11408349 TI - Glutathione-S-transferase gene polymorphisms in colorectal cancer patients: interaction between GSTM1 and GSTM3 allele variants as a risk-modulating factor. AB - The distribution of polymorphisms in the glutathione S-transferase (GST) family genes has been studied in 355 healthy controls and 206 cancer (59 proximal and 147 distal) patients. All controls were subjected to flexible sigmoidoscopy. Odds ratios (OR) after stratification by age, gender and smoking were slightly higher in the cancer group as a whole for GSTM1-null (*0/*0), GSTT1-null (*0/*0) and GSTM3 *A/*B or *B/*B when compared with the control group, but the differences did not reach statistical significance. GSTP1 variants had no effect. Separate analysis of patients with proximal and distal tumours has shown stronger associations for the distal cancers, the GSTM3*B allele presence being significantly more frequent in these patients [OR = 1.77; 95% confidence interval (CI) = 1.15-2.74]. Taking into account strong linkage between the GSTM1*A and GSTM3*B alleles, a separate analysis of the GSTM1-nulled individuals was undertaken. The combination of GSTM1-null genotype with GSTM3*B allele presence (*A/*B or *B/*B) was significantly overrepresented among patients with proximal and distal tumours taken together (OR = 2.12; 95% CI = 1.24-3.63), and especially in distal cancer patients (OR = 2.75; 95% CI = 1.56-4.84). Male individuals displayed a stronger association between the presence of the GSTM1-null in combination with GSTM3 *A/*B or *B/*B and distal tumours with a higher odds ratio (OR = 3.57; 95% CI = 1.73-7.36). In contrast, the frequency of GSTM1 *B/*0 or *B/*B combined with GSTM3 *A/*A was significantly lower in patients with distal colorectal cancer, especially in males (OR = 0.37; 95% CI = 0.15-0.92). Neither of these combinations was associated with proximal tumours. Our findings suggest that interactions of polymorphic genotypes within the GSTM gene cluster affect individual susceptibility to colorectal carcinogenesis, the GSTM3*B variant presence being a risk factor especially in combination with the GSTM1-null genotype. PMID- 11408350 TI - HMG-CoA reductase inhibitor mevastatin enhances the growth inhibitory effect of butyrate in the colorectal carcinoma cell line Caco-2. AB - Mevastatin is an inhibitor of 3-hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in cholesterol synthesis. Butyrate, a short chain fatty acid, reduces proliferation and induces differentiation of human colon cancer cells. The aim of our study was to determine the effect of mevastatin, alone or in combination with butyrate, on proliferation, the cell cycle and apoptosis in the human colorectal carcinoma cell line Caco-2. In this report we show that mevastatin combined with butyrate synergistically suppressed growth of Caco-2 cells in a dose- and time-dependent manner. In addition, incubation with mevastatin arrested cells in the G1 phase of the cell cycle after 24 h with a switch to the G2/M phase after 72 h. This was accompanied by a down regulation of cyclin-dependent kinases (cdk) 4 and cdk 6 as well as cyclin D1, while cdk 2 and cyclin E protein levels remained unchanged during mevastatin treatment. Cell cycle inhibitors p21 and p27 were significantly upregulated by mevastatin. The proapoptotic properties of mevastatin were further enhanced by co incubation with butyrate. Lastly, the effects of mevastatin could be reversed by addition of mevalonate, but not farnesyl- or geranylgeranylpyrophosphate, intermediate products of cholesterol synthesis, to the medium. These results suggest that HMG-CoA reductase inhibitors like mevastatin may enhance the antiproliferative effect of butyrate in colon cancer cells via induction of apoptosis together with a G0/G1 cell cycle arrest. PMID- 11408351 TI - Quercetin, E7 and p53 in papillomavirus oncogenic cell transformation. AB - Bovine papillomavirus type 4 (BPV-4) infects the upper alimentary canal of cattle causing benign papillomas which can progress to squamous carcinomas in cattle grazing on bracken fern (BF). We have previously shown that quercetin, a well characterized and potent mutagen found in BF, causes cell cycle arrest of primary bovine cells (PalF), but that a single exposure to quercetin can cause full oncogenic transformation of PalF cells partially transformed by BPV-4. Here we show that cell cycle arrest correlates with an increase in p53 protein levels and transcriptional activity. However, in cells transformed but non-tumorigenic, p53 protein is elevated and transcriptionally activated in response to quercetin or other DNA damaging stimuli, but the cells bypass quercetin-induced G1 arrest likely due to E7 expression. In transformed tumorigenic cells, p53 is elevated in response to quercetin but its transcriptional activity is inhibited due to mutation, and the cells fail to stop in G1 in the presence of quercetin. PMID- 11408352 TI - Effect of a complex environmental mixture from coal tar containing polycyclic aromatic hydrocarbons (PAH) on the tumor initiation, PAH-DNA binding and metabolic activation of carcinogenic PAH in mouse epidermis. AB - Human exposure to polycyclic aromatic hydrocarbons (PAH) occurs through complex mixtures such as coal tar. The effect of complex PAH mixtures on the activation of carcinogenic PAH to DNA-binding derivatives and carcinogenesis were investigated in mice treated topically with NIST (National Institute of Standards and Technology) Standard Reference Material 1597 (SRM), a complex mixture of PAH extracted from coal tar, and either additional benzo[a]pyrene (B[a]P) or dibenzo[a,l]pyrene (DB[a,l]P). In an initiation-promotion study using 12-O tetradecanoylphorbol-13-acetate as the promoter for 25 weeks, the SRM and B[a]P co-treated mice had a similar incidence of papillomas per mouse compared with the group exposed to B[a]P alone as the initiator. PAH-DNA adduct analysis of epidermal DNA by 33P-post-labeling and reversed-phase high-performance liquid chromatography found the SRM co-treatment led to a significant decrease in the total level of DNA adducts and B[a]P-DNA adducts to less than that observed in mice treated with B[a]P alone at 6, 12 and 72 h exposure. After 24 and 48 h exposure, there was no significant difference in the levels of adducts between these groups. In the DB[a,l]P initiation-promotion study, the co-treated group had significantly fewer papillomas per mouse than mice treated with DB[a,l]P alone as initiator. Averaging over the times of exposure gave strong evidence that mice co-treated with SRM and DB[a,l]P had a significantly lower level of PAH DNA adducts than mice treated with DB[a,l]P alone. Western immunoblots showed that both cytochrome P450 (CYP) 1A1 and 1B1 were induced by the SRM. These results are consistent with the hypothesis that two major factors determining the carcinogenic activity of PAH within a complex mixture are (i) the persistence of certain PAH-DNA adducts as well as total adduct levels, and (ii) the ability of the components present in the mixture to inhibit the activation of carcinogenic PAH by the induced CYP enzymes. PMID- 11408353 TI - N-glucuronidation of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and N_hydroxy-PhIP by specific human UDP-glucuronosyltransferases. AB - Glucuronidation is a major metabolic pathway in the biotransformation of many xenobiotics. Recent studies have shown that in humans, UDP glucuronosyltransferase (UGT)-mediated glucuronidation plays a critical role in the detoxification of food-borne carcinogenic heterocyclic amines. 2-Amino-1 methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), the most abundant carcinogenic heterocyclic amine found in well-cooked meats, has been shown to be extensively glucuronidated in humans. To determine which UGT isozymes are involved in the biotransformation of PhIP and the cytochrome P4501A2-mediated reactive intermediate N-hydroxy-PhIP, microsomes expressing human UGT1A1, -1A4, -1A6 or 1A9 were incubated with PhIP and N-hydroxy-PhIP and the reaction products analyzed by HPLC and ESI-MS. Incubations containing N-hydroxy-PhIP and UGT1A1 expressing microsomes, with an apparent Km of 4.58 microM and a Vmax of 4.18 pmol/min/mg protein, had the highest capacity to convert N-hydroxy-PhIP to N hydroxy-PhIP-N2-glucuronide. Microsomes expressing UGT1A9 produced N-hydroxy-PhIP N3-glucuronide at the highest rate with an apparent Km and Vmax of 3.73 microM and 4.07 pmol/min/mg, respectively. A third previously undefined glucuronide accounted for 31% of the total glucuronides formed from the UGT1A4 expressing microsomes. No glucuronide conjugates were detected from microsomes expressing UGT1A6. Incubations containing PhIP as substrate formed direct PhIP-glucuronides in microsomes expressing UGT1A1, UGT1A4 and UGT1A9 but at levels averaging 53 fold lower than when N-hydroxy-PhIP was used as the substrate. Knowing the glucuronidation capacity of the specific UGT isozymes involved in PhIP and N hydroxy-PhIP glucuronidation should help in determining the individual susceptibility to the potential cancer risk from exposure to PhIP. PMID- 11408354 TI - Urinary excretion of N-OH-2-amino-3-methylimidazo[4,5-f]quinoline-N-glucuronide in F344 rats is enhanced by green tea. AB - The effects of green tea on the metabolism of the food carcinogen 2-amino-3 methylimidazo[4,5-f]quinoline (IQ) with emphasis on the formation of the detoxified glucuronides was studied. Two groups of 20 adult male and female Fischer 344 rats consumed 2% green tea or water for 6 weeks before being administered a single dose of 40 mg/kg body weight of [2-14C]IQ by oral gavage. Major metabolites in 24 h urine samples were separated by high-performance liquid chromatography (HPLC), including N-OH-IQ-N-glucuronide, 5-OH-IQ glucuronide and sulfate, IQ sulfamate and IQ itself. The structures of the main metabolites were established by mobility on the HPLC and by mass spectrometry. Sulfate esters and sulfamate were hydrolyzed by 0.1 N HCl for 15 min at 100 degrees C, yielding 5-OH IQ and high levels of IQ. HPLC of the resulting product showed the N-OH-IQ-N glucuronide and the 5-OH-IQ glucuronide, as well as IQ. The male and female rats drinking tea displayed a significantly higher (P < 0.05) excretion of the two major glucuronides. We conclude that intake of green tea increases the excretion of N-OH-IQ-N-glucuronide, a detoxified metabolite of the proximate carcinogen N OH-IQ. PMID- 11408355 TI - DMBA-induced toxic and mutagenic responses vary dramatically between NER deficient Xpa, Xpc and Csb mice. AB - Heterogeneity in cancer susceptibility exists between patients with an inherited defect in nucleotide excision repair (NER). While xeroderma pigmentosum (XP) patients have elevated skin cancer rates, Cockayne syndrome (CS) patients do not appear to have increased cancer susceptibility. To investigate whether differences in mutagenesis are the basis for the variability in cancer proneness, we studied mutagenesis at the X-chromosomal Hprt gene and the autosomal Aprt gene in splenic T-lymphocytes after 7,12-dimethyl-1,2-benz[a]anthracene (DMBA) exposure in total NER-deficient Xpa mice, global genome repair (GGR)-deficient Xpc mice and transcription coupled repair (TCR)-deficient Csb mice. Surprisingly, while all intraperitoneally-treated Xpc(-/-) mice survived a dose of 40 mg/kg DMBA, a substantial fraction of the treated Xpa(-/-) and Csb(-/-) mice died a few days after treatment with a 20-fold lower dose. Functional TCR of DMBA adducts in Xpc(-/-) mice thus appears to alleviate DMBA toxicity. However, the mutagenic response in Xpc(-/-) mice was +/- 2-fold enhanced at both the Hprt and the Aprt gene compared to heterozygous controls, indicating that GGR at least partially removes DMBA adducts from the genome overall. DMBA-induced SCE frequencies in mouse dermal fibroblasts were significantly enhanced in Xpa- and Csb-, but not in Xpc-deficient background compared to the frequency in normal fibroblasts. These results indicate that both damage-induced cytotoxicity as well as intra chromosomal recombinational events were not correlated to differences in cancer susceptibility in human NER syndrome patients. PMID- 11408356 TI - Cassette dosing pharmacokinetics: valuable tool or flawed science? PMID- 11408357 TI - Pharmacophore and three-dimensional quantitative structure activity relationship methods for modeling cytochrome p450 active sites. AB - Structure activity relationships (SAR), three-dimensional structure activity relationships (3D-QSAR), and pharmacophores represent useful tools in understanding cytochrome P450 (CYP) active sites in the absence of crystal structures for these human enzymes. These approaches have developed over the last 30 years such that they are now being applied in numerous industrial and academic laboratories solely for this purpose. Such computational approaches have helped in understanding substrate and inhibitor binding to the major human CYPs 1A2, 2B6, 2C9, 2D6, 3A4 as well as other CYPs and additionally complement homology models for these enzymes. Similarly, these approaches may assist in our understanding of CYP induction. This review describes in detail the development of pharmacophores and 3D-QSAR techniques, which are now being more widely used for modeling CYPs; the review will also describe how such approaches are likely to further impact our active site knowledge of these omnipresent and important enzymes. PMID- 11408358 TI - Pharmacokinetic studies of 2-amino-9-(3-acetoxymethyl-4-isopropoxycarbonyl-oxybut 1-yl)purine, an oral prodrug for the antiviral agent penciclovir. AB - 2-Amino-9-(3-acetoxymethyl-4-isopropoxycarbonyloxybut-1-yl)- purine (SK1899) was tested as an oral prodrug for penciclovir. SK1899 was administered orally to rats and dogs at doses up to 2 and 0.68 mmol/kg, respectively. SK1899 was well absorbed, and the major metabolites detected in plasma and urine were penciclovir, the active antiviral compound, and 6-deoxypenciclovir (M4) in both species. In rats, SK1899 was rapidly and extensively metabolized to penciclovir, which reached the peak plasma concentration (C(max)) of 39.5 microM at 0.5 h after 0.2-mmol/kg dosing. The area under the plasma concentration-time curve (AUC) for penciclovir was 57.5 microM x h. After an oral dose of 0.034 mmol/kg to dogs, extensive conversion of SK1899 to penciclovir also occurred with slower rate of formation of penciclovir from M4 than in rats. The mean C(max) and AUC for penciclovir were 4.5 microM at 2.7 h and 28.2 microM x h, respectively. The 0 to 24-h urinary recovery of penciclovir represented 36.1 and 36.3% of dose to rats and dogs, respectively. Radioactivity was found in fetuses following an oral administration of [(14)C]SK1899 to pregnant rats, but no significant accumulation was observed. Although substantial milk transfer of [(14)C]SK1899 occurred in rats, the radioactivity in milk was rapidly cleared. The values of C(max), AUC, and urinary recovery of penciclovir after dosing with SK1899 to rats and dogs were similar or slightly higher than those from famciclovir. These data indicate that introduction of an isopropoxy carbonate group into one of the two hydroxyl groups of M4 did not significantly alter the oral bioavailability of penciclovir compared with famciclovir. PMID- 11408359 TI - Selective dehydrogenation/oxygenation of 3-methylindole by cytochrome p450 enzymes. AB - 3-Methylindole (3 MI) is a selective pulmonary toxicant, and cytochrome P450 (P450) bioactivation of 3 MI, through hydroxylation, epoxidation, or dehydrogenation pathways, is a prerequisite for toxicity. CYP2F1 and CYP2F3 exclusively catalyze the dehydrogenation of 3 MI to 3-methyleneindolenine, without detectable formation of the hydroxylation or epoxidation products. It was not known whether 3 MI is simply an excellent dehydrogenation substrate for all P450 enzymes, or whether certain cytochrome P450s responsible for 3 MI bioactivation have unique active sites that only catalyze the dehydrogenation of the molecule, while other P450s would catalyze only the oxygenation of 3 MI. Therefore, the kinetics of product formation by the CYP2F1 and CYP2F3 enzymes were compared with other cytochrome P450 enzymes. The enzymes tested were CYP1A1, CYP1A2, CYP1B1, and CYP2E1. The CYP1A1 and CYP1A2 enzymes produced all three 3 MI metabolites: the dehydrogenation product, 3-methyleneindolenine (V(max)/K(m) = 4 and 22, respectively); the hydroxylation product, indole-3-carbinol (V(max)/K(m) = 42 and 100, respectively); and the epoxidation product, 3-methyloxindole (V(max)/K(m) = 4 and 72, respectively). These CYP1A enzymes catalyzed oxygenation of 3 MI at much faster rates than dehydrogenation. CYP1B1 produced indole-3 carbinol (V(max)/K(m) = 85) and 3-methyloxindole (V(max)/K(m) = 7), and CYP2E1 only produced 3-methyloxindole (V(max)/K(m) = 98), but neither enzyme catalyzed the formation of the dehydrogenated product. Six additional P450 enzymes that were tested formed none of the dehydrogenation product. The ability of the various CYP1 family enzymes to catalyze the formation of all three major 3 MI metabolites, along with the specific oxygenation by CYP2E1, illustrates that dehydrogenation of 3 MI is not a substrate-directed process, but that the members of the CYP2F family possess unique active sites that specifically catalyze only the dehydrogenation mechanism. PMID- 11408360 TI - Methadone inhibits rhodamine123 transport in Caco-2 cells. AB - This study investigated the effects of racemic methadone (MET) on P-glycoprotein (P-gp) activity in cell culture. MET showed no differential rates of passage between the basolateral to apical (B to A) and apical to basolateral (A to B) direction across Caco-2 cell monolayers in a transwell system. MET transport in either direction was not importantly influenced by the P-gp inhibitor verapamil. However, MET was a potent inhibitor (IC(50) = 7.5 microM) of rhodamine123 B to A transport across Caco-2 cell monolayers, causing a reduction to 25% of control at 100 microM MET. In this model of Caco-2 monolayers, rates of MET passage between B to A and A to B directions could not be distinguished. However, MET can inhibit P-gp activity at intraluminal concentrations that might be achieved clinically. This may lead to increased bioavailability of coadministered compounds. PMID- 11408361 TI - Pharmacokinetic theory of cassette dosing in drug discovery screening. AB - Cassette dosing is a procedure for higher-throughput screening in drug discovery to rapidly assess pharmacokinetics of large numbers of candidate compounds. In this procedure, multiple compounds are administered simultaneously to a single animal. Blood samples are collected, and the plasma samples obtained are analyzed by means of an assay method such as liquid chromatography coupled to tandem mass spectrometry that permits concurrent assay of many compounds in a single sample. Consequently, the pharmacokinetics of multiple compounds can be assessed rapidly with a small number of experimental animals and with shortened assay times. However, coadministration of multiple compounds may result in pharmacokinetic drug-drug interactions. This paper describes a pharmacokinetic description for cassette dosing derived from pharmacokinetic theory. The most important finding from this theoretical treatment is that the potential for drug-drug interactions leading to altered clearances of coadministered drugs depends on both the relative K(M) values for the metabolic enzymes and the total number of drugs coadministered. However, the theory predicts that the potential for drug-drug interactions is only a weak function of the dose size. Finally, it is also shown that including a benchmark compound within the set of coadministered compounds cannot ensure the detection of errors due to drug-drug interactions. Thus, neither the absolute values of pharmacokinetic parameters nor the rank order obtained from cassette dosing can be accepted without independent confirmation. These theoretical predictions are evaluated with data taken from the literature. PMID- 11408362 TI - Influence of dose and infusion duration on pharmacokinetics of ifosfamide and metabolites. AB - The anticancer drug ifosfamide is a prodrug requiring activation through 4 hydroxyifosfamide to ifosforamide mustard, to exert cytotoxicity. Deactivation of ifosfamide leads to 2- and 3-dechloroethylifosfamide and the release of potentially neurotoxic chloracetaldehyde. The aim of this study was to quantify and to compare the pharmacokinetics of ifosfamide, 2- and 3 dechloroethylifosfamide, 4-hydroxyifosfamide, and ifosforamide mustard in short (1-4 h), medium (24-72 h), and long infusion durations (96-240 h) of ifosfamide. An integrated population pharmacokinetic model was used to describe the autoinducible pharmacokinetics of ifosfamide and its four metabolites in 56 patients. The rate by which autoinduction of the metabolism of ifosfamide developed was found to be significantly dependent on the infusion schedule. The rate was 52% lower with long infusion durations compared with short infusion durations. This difference was, however, comparable with its interindividual variability (22%) and was, therefore, considered to be of minor clinical importance. Autoinduction caused a less than proportional increase in the area under the ifosfamide plasma concentration-time curve (AUC) and more than proportional increase in metabolite exposure with increasing ifosfamide dose. During long infusion durations dose-corrected exposures (AUC/D) were significantly decreased for ifosfamide and increased for 3 dechloroethylifosfamide compared with short infusion durations. No differences in dose-normalized exposure to ifosfamide and metabolites were observed between short and medium infusion durations. This study demonstrates that the duration of ifosfamide infusion influences the exposure to the parent and its metabolite 3 dechloroethylifosfamide. The observed dose and infusion duration dependence should be taken into account when modeling ifosfamide metabolism. PMID- 11408363 TI - Metabolism of norethisterone and norethisterone derivatives in rat uterus, vagina, and aorta. AB - The 19-nor-progestogen norethisterone is used as a progestogen component in contraceptives and in continuous- and sequential combined hormone replacement therapy (HRT) in postmenopausal women. Metabolism of norethisterone in HRT target tissues may play a role in its biological response. The aim of this study was to investigate which steroid-metabolizing enzymes are present in rat uterus, vagina, and aorta, three HRT target tissues. Next, the ability of the tissues to metabolize norethisterone was assessed. Furthermore, to investigate the effect of substituents at the 7- and 11-position, the metabolism of Org OM38 (7alpha-methyl norethisterone), Org 4060 (11beta-ethyl-norethisterone), and Org 34694 (7alpha methyl,11-ethylidene-norethisterone) was studied. Using radiolabeled progesterone, the presence of 20alpha-hydroxysteroid dehydrogenase, 5alpha reductase, and 3alpha-hydroxysteroid dehydrogenase activity could be demonstrated in uterus, vagina, and to a lesser extent in aorta. The combined action of the latter two enzyme activities resulted in 3alpha-OH,5alpha-H-norethisterone as the major metabolite of radiolabeled norethisterone in uterus (26.9%), vagina (37.1%), and aorta (1.4%). The norethisterone derivatives, however, were metabolized to a much lesser extent (1.0-7.6%). No formation of 5alpha-reduced forms of Org 4060, Org OM38, or Org 34694 was found, while formation of minor amounts of 3alpha-OH-Org 4060 and 3alpha-OH-Org OM38 could be demonstrated in both uterus, vagina, and aorta. These findings confirm the role of 5alpha reductase as a rate-limiting step in the metabolism of norethisterone derivatives and show important inhibitory effects of substituents at the 7alpha- and 11 position of the steroid skeleton on 5alpha-reduction. PMID- 11408364 TI - Screening of organosulfur compounds as inhibitors of human CYP2A6. AB - The capacities to inhibit coumarin 7-hydroxylase activity of human cytochrome P450 2A6 (CYP2A6) by organosulfur compounds were evaluated. Five dialkyl sulfides and five dialkyl disulfides, with alkyl chains from methyl to amyl, were examined. In addition to these chemicals, diallyl sulfide, diallyl disulfide, allyl methyl sulfide, allyl n-propyl sulfide, allyl phenyl sulfide, diphenyl sulfide, diphenyl disulfide, difurfuryl disulfide, phenyl cyclopropyl sulfide, 2,2'-dipyridyl disulfide, 4,4'-dipyridyl sulfide, and 4,4'-dipyridyl disulfide were also examined for their capacity to inhibit CYP2A6. The membrane fraction of genetically engineered Escherichia coli cells expressing CYP2A6 together with NADPH-cytochrome P450 reductase was used as an enzyme source. Dialkyl disulfides inhibited CYP2A6 more strongly than did dialkyl sulfides. Among dialkyl disulfides examined, di-n-propyl disulfide, contained in onion oil, was the most potent competitive inhibitor of CYP2A6, with a K(i) value of 1.73 microM. Diallyl disulfide, present in garlic oil, inhibited CYP2A6 activity in a competitive/noncompetitive mixed manner, with the K(i) value of 2.13 microM. Among all of the organosulfur compounds tested, 4,4'-dipyridyl disulfide was the most potent inhibitor of CYP2A6, with a K(i) value of 60 nM, followed by 4,4' dipyridyl sulfide, with a K(i) value of 72 nM. These chemicals inhibited CYP2A6 in a competitive manner. The preincubation time did not affect the inhibitory effects of di-n-propyl disulfide, diallyl disulfide, 4,4'-dipyridyl disulfide, and 4,4'-dipyridyl sulfide on CYP2A6, indicating that these chemicals were not mechanism-based inhibitors of CYP2A6. 4,4'-Dipyridyl disulfide also inhibited midazolam 1'-hydroxylase activity of CYP3A4. We discovered 4,4'-dipyridyl disulfide to be a potent and relatively selective inhibitor of CYP2A6. PMID- 11408365 TI - Bexarotene metabolism in rat, dog, and human, synthesis of oxidative metabolites, and in vitro activity at retinoid receptors. AB - The metabolism of bexarotene, a rexinoid recently approved in the United States for treatment of cutaneous T-cell lymphoma, was studied using liver slices from untreated rats and dogs, liver microsomes from untreated and pretreated rats, and pooled human liver microsomes. Metabolite profiles were examined in bile and plasma from rats and dogs, and plasma from humans treated with bexarotene. Four metabolites, racemic 6-hydroxy-bexarotene, racemic 7-hydroxy-bexarotene, 6-oxo bexarotene, and 7-oxo-bexarotene, were synthesized and their binding to, and transactivation of retinoid receptors were examined. Qualitatively similar metabolite profiles were observed in the microsomal and liver slice extracts; the predominant metabolites were 6-hydroxy-bexarotene and glucuronides of parent or hydroxylated metabolites. Pretreatment of rats with bexarotene induced hepatic microsomal bexarotene metabolism. The hydroxy and oxo metabolites were observed in plasma of rats, dogs, and humans treated with bexarotene and 6-hydroxy bexarotene was a major circulating metabolite. The oxidative metabolites were more abundant relative to parent in plasma from humans than from rat or dog. The predominant biliary metabolites in rat and dog were bexarotene acyl glucuronide and a glucuronide of oxidized bexarotene, respectively. Since bexarotene elimination is primarily biliary in these species, these metabolites represent the main bexarotene metabolites in rats and dogs. The binding of synthetic metabolites to retinoid receptors was much reduced relative to parent compound. The metabolites exhibited minimal activity in transactivating retinoic acid receptors and had reduced activity at retinoid X receptors relative to bexarotene. Thus, while there is substantial systemic exposure to the oxidative metabolites of bexarotene, they are unlikely to elicit significant retinoid receptor activation following bexarotene administration. PMID- 11408366 TI - Polycyclic aromatic hydrocarbon/metal mixtures: effect on PAH induction of CYP1A1 in human HEPG2 cells. AB - Environmental polycyclic aromatic hydrocarbons (PAHs) and metals coexist, and such mixtures could affect the carcinogenicity of PAHs, possibly by modification of PAH induction of the PAH-bioactivating CYP1A. The effect on PAH-mediated CYP1A induction of arsenic, lead, mercury, or cadmium (ranked as the most hazardous environmental metals by the Environmental Protection Agency and the Agency for Toxic Substances and Disease Registry) has thus been investigated. Induction of CYP1A1 by benzo[a]pyrene (BAP), benzo[b]fluoranthene (BBF), dibenzo[a,h]anthracene (DBAHA), benzo[a]anthracene (BAA), or benzo[k]fluoranthene (BKF) was probed by ethoxyresorufin-O-deethylase activity (EROD) in 96-well plates of human HepG2 cells, by immunoblot analysis, and by reverse transcription polymerase chain reaction. Cells rapidly took up PAHs (BAP, BKF) from medium; by 24 h only 14% remained in the medium, and no detectable PAH bound to well walls. Induction efficiency (relative to dimethyl sulfoxide controls) was in the order BKF (16-fold) > DBAHA (14-fold) > BAA (4-fold) > BAP (3-fold) > BBF (1-fold), all at 5 microM PAH. The metals did not markedly affect cell viability at concentrations of arsenic, 5 microM; lead, 50 microM; mercury, 5 microM; and cadmium, 5 microM. At 5 microM PAH concentration, all of the metals decreased levels of PAH-induced CYP1A1 activities (direct inhibition of EROD activity was excluded) by variable extents and in a PAH-dependent manner. With BAP as inducer decreases in induction were arsenic, 57%; cadmium, 82%; mercury, 4%; and lead, 20%. The decreases were not a consequence of transcriptional down-regulation. One possible conclusion is that these metals could diminish PAH carcinogenic potential by decreasing PAH-mediated induction of their bioactivation by CYP1A1. PMID- 11408367 TI - Cytochrome P450 down-regulation by serum from humans with a viral infection and from rabbits with an inflammatory reaction. AB - Serum from humans with an upper respiratory viral infection (HS(URVI)) and from rabbits with a turpentine-induced acute inflammatory reaction (RS(TIAR)) reduces the activity of hepatic cytochrome P450 (P450) following 4 h of incubation. The aim of the present study was to assess the effect of HS(URVI) and RS(TIAR) on P450 activity and expression following 24 h of incubation with hepatocytes from control (H(CONT)) and rabbits with a TIAR (H(INFLA)). RS(TIAR) incubated with H(CONT) for 24 h reduced P450 content and activity, and CYP3A6 by 45%, without changing CYP1A1 and 1A2; when incubated with H(INFLA), RS(TIAR) decreased P450 content and activity without affecting CYP1A1 or 1A2. HS(URVI) incubated for 4 h with H(CONT) decreased P450 activity without affecting the amounts of CYP1A1, 1A2, or 3A6, although when incubated for 24 h, P450 activity and CYP3A6 amount decreased. HS(URVI) incubated with H(INFLA) for 4 h reduced P450 content and activity, and incubated for 24 h reduced activity, P450 content, and amount of CYP1A1 and 1A2 proteins. The present study demonstrates that 1) the effect of RS(TIAR) and HS(URVI) depends upon the susceptibility of the hepatocyte, i.e., H(CONT) or primed H(INFLA); 2) P450 down-regulation is preceded by a decrease in P450 activity; 3) the nature of the inflammatory reaction determines the repercussions on P450 activity and expression; and 4) CYP3A6 is more vulnerable than CYP1A1 and 1A2 to the down-regulation provoked by an inflammatory challenge. PMID- 11408368 TI - Dietary uptake kinetics of 2,2',5,5'-tetrachlorobiphenyl in rainbow trout. AB - The disposition of [UL-(14)C]2,2',5,5'-tetrachlorobiphenyl (TCB) in rainbow trout (Oncorhynchus mykiss) was studied in acute dietary exposures using TCB contaminated fathead minnows (Pimephales promelas). Trout were sampled at several postfeeding time points and TCB-derived radioactivity was measured in gut contents and selected tissues. Gastric evacuation was exponential with time and was 95% complete within 36 h of feeding. The ratio of activity in upper intestinal tissue to that in blood declined between 6 and 48 h, as did the lumenal contents/tissue ratio. Stomach content lipid declined between 0 and 24 h, while the lipid content of chyme remained relatively constant. These observations are consistent with liquid phase emptying of lipid and TCB to the upper intestine followed by rapid coassimilation. Tissue/blood activity ratios for the stomach, lower intestine, muscle, liver, and kidney were constant and probably represented near equilibrium conditions. The fat/blood activity ratio increased through 96 h, indicating that TCB was redistributing to fat. The lower intestinal tissue/feces activity ratio increased between 6 and 24 h and then declined rapidly. Fecal lipid content also increased between 6 and 24 h, but the amount of this increase was insufficient to explain observed changes in the distribution of TCB-derived activity. A small amount of 3-hydroxy TCB was detected in feces. Generally, however, metabolism had little or no impact on the uptake, distribution or elimination of TCB. Measured assimilation efficiencies exceeded 90% and are the highest ever reported in fish feeding studies with TCB. PMID- 11408369 TI - In vivo effect of clarithromycin on multiple cytochrome P450s. AB - The in vivo effects of oral clarithromycin administration on the in vivo activity of cytochrome P450 1A2, 2C9, and 2D6 were determined. The cytochrome P450 probes caffeine (CYP1A2), tolbutamide (CYP2C9), and dextromethorphan (CYP2D6) were administered as an oral cocktail prior to and 7 days after oral clarithromycin (500 mg twice daily) administration to 12 healthy male subjects. Blood and urine samples were collected and assayed for each of the compounds and their metabolites using high-performance liquid chromatography. The CYP1A2 indices, oral caffeine clearance (6.2 +/- 3.3 l/h before and 5.7 +/- 4.2 l/h after, p > 0.05) and the 6-h paraxanthine to caffeine serum concentration ratio (0.49 +/- 0.3 before and 0.44 +/- 0.3 after, p > 0.05), were unchanged following clarithromycin dosing. Neither the tolbutamide oral clearance (0.77 +/- 0.28 l/h before and 0.72 +/-0.24 l/h after, p > 0.05) nor the tolbutamide urinary metabolic ratio (779 +/- 294 before and 681 +/- 416 after, p > 0.05) indices of CYP2C9 were altered by clarithromycin administration. In the case of CYP2D6, the dextromethorphan to dextrorphan urinary ratio was not significantly different before (0.021 +/- 0.04) and after (0.024 +/- 0.06) clarithromycin dosing. In conclusion, clarithromycin does not appear to alter the in vivo catalytic activity of CYP1A2, CYP2C9, and CYP2D6 in healthy individuals as assessed by caffeine, tolbutamide, and dextromethorphan, respectively. PMID- 11408370 TI - Dapsone activation of CYP2C9-mediated metabolism: evidence for activation of multiple substrates and a two-site model. AB - Dapsone activates CYP2C9-mediated metabolism in various expression systems and is itself metabolized by CYP2C9 to its hydroxylamine metabolite. Studies were conducted with expressed CYP2C9 to characterize the kinetic effects of dapsone (0 100 microM) on (S)-flurbiprofen (2-300 microM), (S)-naproxen (10-1800 microM), and piroxicam (5-900 microM) metabolism in 6 x 6 matrix design experiments. The influence of (S)-flurbiprofen on dapsone hydroxylamine formation was also studied. Dapsone increased the Michaelis-Menten-derived V(max) of flurbiprofen 4' hydroxylation from 12.6 to 20.6 pmol/min/pmol P450, and lowered its K(m) from 28.9 to 10.0 microM, suggesting that dapsone activates CYP2C9-mediated flurbiprofen metabolism without displacing flurbiprofen from the active site, supporting a two-site model describing activation. Similar results were observed with piroxicam 5'-hydroxylation, as V(max) was increased from 0.08 to 0.20 pmol/min/pmol P450 and K(m) was decreased from 183 to 50 microM in the presence of dapsone. In addition, the kinetic profile for naproxen was converted from biphasic to hyperbolic in the presence of dapsone, while exhibiting similar decreases in K(m) and increases in V(max). Kinetic parameters were also estimated using the two-site binding equation, with alpha values <1 and beta values >1, indicative of activation. Additionally, dapsone hydroxylamine formation was measured from incubations containing flurbiprofen, exhibiting a kinetic profile that was minimally affected by the presence of flurbiprofen. Overall, these results suggest that dapsone activates the metabolism of multiple substrates of CYP2C9 by binding within the active site and causing positive cooperativity, thus lending further support to a two-site binding model of P450-mediated metabolism. PMID- 11408371 TI - In vivo pharmacokinetics and metabolism of anti-human immunodeficiency virus agent D4T-5'-[p-bromophenyl methoxyalaninyl phosphate] (SAMPIDINE) in mice. AB - d4T-5'-[p-Sampidine, bromophenyl methoxyalaninyl phosphate] (HI-113), a novel aryl phosphate derivative of stavudine (d4T), exhibits substantially more potent anti-human immunodeficiency virus activity than d4T. The purpose of the present study was to investigate the in vivo pharmacokinetics and metabolism of this promising new anti-HIV agent in mice. Here, we report that HI-113 forms two active metabolites with favorable pharmacokinetics after systemic administration. Plasma HI-113 concentrations were measured by analytical high-performance liquid chromatography and the pharmacokinetic parameters were estimated using the WinNonlin program. After intravenous administration, the elimination half-life (t(1/2)) of HI-113 was 3.6 min with a systemic clearance of 174.5 ml/min/kg. HI 113 was converted to the active metabolites alaninyl-d4T-monophosphate (ala-d4T MP) and d4T. The T(max) values for ala-d4T-MP and d4T derived from intravenously administered HI-113 were 5.1 and 17.4 min, respectively. The elimination half life for synthetic ala-d4T-MP was 38.9 min after intravenous administration. Ala d4T-MP was metabolized to form d4T (T(max) = 5.0 min). The elimination half-life of d4T derived from intravenously administered ala-d4T-MP (32.4 min) was similar to the elimination half-life of intravenously administered d4T (26.6 min). In contrast, the elimination half-life of d4T derived from HI-113 was substantially longer (116.2 min). Similarly, the elimination half-life of ala-d4T-MP derived from HI-113 (138.8 min) was markedly longer than the elimination half-life of ala d4T-MP given intravenously (38.9 min). Following oral administration of HI-113, the elimination half-lives of ala-d4T-MP (56.1 min) and d4T (102.6 min) were also prolonged. PMID- 11408372 TI - Studies comparing in vivo:in vitro metabolism of three pharmaceutical compounds in rat, dog, monkey, and human using cryopreserved hepatocytes, microsomes, and collagen gel immobilized hepatocyte cultures. AB - The in vivo metabolism of three pharmaceutical compounds, EMD68843, EMD96785, and EMD128130, was compared in fresh and cryopreserved hepatocyte (CPH) suspensions and microsomes from rat, dog, monkey, and human livers and fresh human and rat hepatocyte collagen gel immobilized cultures (GICs). Half of the major in vivo metabolites was produced by phase 1 (hydroxylation, oxidation, hydrolysis, N dealkylation) and half by phase 2 metabolism (mostly glucuronidation but also sulfation and glycine conjugation). The identity and percentage of phase 1 and 2 metabolites from each compound produced in hepatocytes compared well with that in each species in vivo. Glucuronidation was more extensive in GICs than in CPHs. In contrast, CPHs but not GICs, produced sulfate metabolites. Microsomes (supplemented with NADPH only) produced most of the phase 1 but no phase 2 metabolites. Metabolism in CPHs was the same as in fresh hepatocyte suspensions. Discrete species differences in metabolism were detected by CPHs and microsomes. Cytochrome P450 and glucuronosyl S-transferase contents of CPHs did not account for species differences in the percentage of phase 1 and 2 metabolites or the rate of disappearance of the parent compounds in these cells. These data show a good correlation between major metabolites formed in vivo and in vitro. CPHs and GICs, unlike microsomes, carried out sequential phase 1 and 2 metabolism. Each in vitro system has its own advantages, however, for short-term metabolism studies CPHs may be more useful since they are readily available, easier and quicker to prepare than GICs, and have more comprehensive enzyme systems than microsomes. PMID- 11408373 TI - Role of CYP2C9 polymorphism in losartan oxidation. AB - Losartan, an angiotensin II receptor antagonist, is oxidized by hepatic cytochromes P450 to an active carboxylic acid metabolite, E-3174. The aim of the present investigation was to study the contribution of CYP2C9 and CYP3A4 in losartan oxidation in vitro and to evaluate the role of CYP2C9 polymorphism. Kinetic properties of different genetic CYP2C9 variants were compared both in a yeast expression system and in 25 different samples of human liver microsomes where all known genotypes of CYP2C9 were represented. Microsomes were incubated with losartan (0.05-50 microM), and the formation of E-3174 was analyzed by high performance liquid chromatography to estimate V(max), K(m), and intrinsic clearance for all individual samples. Sulfaphenazole, a CYP2C9 inhibitor, blocked the formation of E-3174 at low losartan concentrations (<1 microM), whereas the inhibitory effect of triacetyloleandomycin, a CYP3A4 inhibitor, was significant only at high concentrations of losartan (>25 microM). In comparison to the CYP2C9.1 variant, oxidation of losartan was significantly reduced in yeast expressing the rare CYP2C9.2 or CYP2C9.3 variants. Moreover, the rate of losartan oxidation was lower in liver microsomes from individuals hetero- or homozygous for the CYP2C9*3 allele, or homozygous for the CYP2C9*2 allele. The difference between the common and rare CYP2C9 variants was mainly explained by a lower V(max), both in yeast and human liver microsomes. In summary, these in vitro results indicate that CYP2C9 is the major human P450 isoenzyme responsible for losartan oxidation and that the CYP2C9 genotype contributes to interindividual differences in losartan oxidation and activation. PMID- 11408374 TI - Cytochrome P450-catalyzed metabolism of ezlopitant alkene (CJ-12,458), a pharmacologically active metabolite of ezlopitant: enzyme kinetics and mechanism of an alkene hydration reaction. AB - Experiments were conducted to characterize the metabolism of ezlopitant alkene (CJ-12,458), an active metabolite of ezlopitant, in human liver microsomes. In incubations with human liver microsomes and cofactors required for cytochrome P450 (CYP) activity, CJ-12,458 was converted to two metabolites: a diol (CP 611,781) and a 1 degrees alcohol (CP-616,762). In human liver microsomes, apparent K(M) values of 5.4 and 8.5 microM were determined for the formation of diol and 1 degrees alcohol metabolites, respectively. High K(M) activities were also observed for formation of these metabolites; however, the aforementioned low K(M) activities accounted for greater than 90% of the total intrinsic clearance. In pooled human liver microsomes, formation of both metabolites was partially inhibited by both quinidine and ketoconazole, suggesting that CYP2D6 and CYP3A enzymes are involved in the metabolism of CJ-12,458. This evidence was corroborated through the use of heterologously expressed CYP enzymes and correlation analysis with a panel of human liver microsomes. The data suggest that CYP2D6 is quantitatively more important than CYP3A in the metabolism of CJ 12,458 by a factor of about 2 to 1. The conversion of an alkene to a 1 degrees alcohol represents a novel biotransformation reaction. Incubations using (18)O(2), (2)H(2)O, [(2)H(5)]CJ-12,458, and [(2)H]NADPH were conducted and the 1 degrees alcohol product was characterized by ion trap mass spectrometry. From these data, a mechanism for this reaction is proposed involving epoxidation, an exocyclic hydride shift, and reduction at the benzylic position. PMID- 11408375 TI - Drug interaction between simvastatin and itraconazole in male and female rats. AB - Taking into account the species and sex differences in drug interactions based on the inhibition of cytochrome P450 (P450)-mediated drug metabolism, we examined whether the interaction between simvastatin and itraconazole observed in humans could also occur in rats, the most commonly used animal species for pharmacokinetic studies. Itraconazole inhibited the in vitro metabolism of simvastatin in female rat liver microsomes, but not in male rat liver microsomes. Using anti-P450 antisera, the main P450 isozyme responsible for the metabolism of simvastatin was identified as CYP3A in female rats and CYP2C11 in male rats. Therefore, the sex difference in the inhibition of simvastatin metabolism by itraconazole seems to be caused by a difference in the P450 isozymes responsible for the metabolism of simvastatin in male and female rats and the different ability of itraconazole to inhibit CYP3A and CYP2C11. In addition, the effect of itraconazole on the pharmacokinetics of simvastatin in rats was also investigated. The area under the curve value of simvastatin was increased approximately 1.6-fold by the concomitant use of itraconazole (50 mg/kg) in female rats, whereas in male rats, itraconazole had no effect. In conclusion, it was found that the results obtained in male rats did not reflect the results in humans as far as the inhibition of simvastatin metabolism by itraconazole was concerned. The P450 isozymes involved in the metabolism of drugs should be taken into consideration when rats are used as a model animal for humans in the investigation of drug interactions. PMID- 11408376 TI - Autonomic function in hypertensive and normotensive subjects: the importance of gender. AB - Baroreceptor reflex sensitivity (BRS) has been found lower and heart rate variability (HRV) parasympathetic markers have been found higher in healthy women than in healthy men. Thus, in the present study we hypothesized gender differences in the autonomic function among hypertensive subjects. Forty-one hypertensive patients and 34 normotensive subjects, age 53+/-1 years, were examined. Four weeks after cessation of antihypertensive therapy, HRV was assessed in 24-hour Holter ECGs, and BRS was calculated with the transfer technique. A t test was performed after log transformation of spectral values. Resting blood pressure and heart rate in the hypertensive and the normotensive groups were 150+/-2/100+/-1 (mean+/-SEM) and 121+/-2/81+/-1 mm Hg, respectively, and 68+/-1 and 60+/-1 bpm, respectively (P<0.0005). Compared with normotensive controls, hypertensive patients had lower total power (1224+/-116 versus 1797+/ 241 ms(2); P=0.03), lower low frequency power (550+/-57 versus 813+/-115 ms(2); P=0.04), lower high frequency power (141+/-23 versus 215+/-38 ms(2); P=0.06), lower root mean square successive difference (28.7+/-2.7 versus 35.7+/-3.0 ms; P=0.03), and PNN50 (4.9+/-0.6% versus 9.8+/-1.5%; P=0.003). BRS was also lower in the hypertensive subjects (7.6+/-0.6 versus 10.4+/-0.8 ms/mm Hg; P=0.005). When comparing the same parameters between normotensive subjects and hypertensive subjects within the same gender group, we found significant reduction (P<0.05) only within the female group. The difference in BRS within the female group was twice that within the male group. Stepwise multiple regression analysis revealed gender, age, HDL cholesterol, and blood pressure as independent explanatory variables of BRS and HRV. Our results suggest that gender is an important determinant of BRS and HRV. Autonomic function parameters were especially impaired in hypertensive women compared with hypertensive men. PMID- 11408377 TI - Animal model of neuropathic tachycardia syndrome. AB - Clinically relevant autonomic dysfunction can result from either complete or partial loss of sympathetic outflow to effector organs. Reported animal models of autonomic neuropathy have aimed to achieve complete lesions of sympathetic nerves, but incomplete lesions might be more relevant to certain clinical entities. We hypothesized that loss of sympathetic innervation would result in a predicted decrease in arterial pressure and a compensatory increase in heart rate. Increased heart rate due to loss of sympathetic innervation is seemingly paradoxical, but it provides a mechanistic explanation for clinical autonomic syndromes such as neuropathic postural tachycardia syndrome. Partially dysautonomic animals were generated by selectively lesioning postganglionic sympathetic neurons with 150 mg/kg 6-hydroxydopamine hydrobromide in male Sprague Dawley rats. Blood pressure and heart rate were monitored using radiotelemetry. Systolic blood pressure decreased within hours postlesion (Delta>20 mm Hg). Within 4 days postlesion, heart rate rose and remained elevated above control levels. The severity of the lesion was determined functionally and pharmacologically by spectral analysis and responsiveness to tyramine. Low frequency spectral power of systolic blood pressure was reduced postlesion and correlated with the diminished tyramine responsiveness (r=0.9572, P=0.0053). The tachycardia was abolished by treatment with the beta-antagonist propranolol, demonstrating that it was mediated by catecholamines acting on cardiac beta receptors. Partial lesions of the autonomic nervous system have been hypothesized to underlie many disorders, including neuropathic postural tachycardia syndrome. This animal model may help us better understand the pathophysiology of autonomic dysfunction and lead to development of therapeutic interventions. PMID- 11408378 TI - Quantification of mechanical and neural components of vagal baroreflex in humans. AB - Traditionally, arterial baroreflex control of vagal neural outflow is quantified by heart period responses to falling and/or rising arterial pressures (ms/mm Hg). However, it is arterial pressure-dependent stretch of barosensory vessels that determines afferent baroreceptor responses, which, in turn, generate appropriate efferent cardiac vagal outflow. Thus, mechanical transduction of pressure into barosensory vessel stretch and neural transduction of stretch into vagal outflow are key steps in baroreflex regulation that determine the conventional integrated input-output relation. We developed a novel technique for direct estimation of gain in both mechanical and neural components of integrated cardiac vagal baroreflex control. Concurrent, beat-by-beat measures of arterial pressures (Finapres), carotid diameters (B-mode ultrasonography), and R-R intervals (ECG lead II) were made during bolus vasoactive drug infusions (modified Oxford technique) in 16 healthy humans. The systolic carotid diameter/pressure relationship (r(2)=0.79+/-0.008, mean+/-SEM) provided a gain estimate of dynamic mechanical transduction of pressure into a baroreflex stimulus. The R-R interval/systolic diameter relationship (r(2)=0.77+/-0.009) provided a gain estimate of afferent-efferent neural transduction of baroreflex stimulus into a vagal response. Variance between repeated measures for both estimates was no different than that for standard gain (P>0.40). Moreover, in these subjects, the simple product of the 2 estimates almost equaled standard baroreflex gain (ms/mm Hg=0.98x+2.27; r(2)=0.93, P=0.001). This technique provides reliable information on key baroreflex components not distinguished by standard assessments and gives insight to dynamic mechanical and neural events during acute changes in arterial pressure. PMID- 11408379 TI - Role of brain angiotensin II on somatosensory-induced antinatriuresis in hypertensive rats. AB - The aim of this investigation was to compare the contribution of brain angiotensin II in mediating the transmission of a somatosensory stimulus within the brain to generate a renal sympathetic nerve-dependent antidiuresis and antinatriuresis in normotensive Wistar rats and stroke-prone spontaneously hypertensive rats (SHRSP). In anesthetized Wistar rats, stimulation of somatosensory receptors by subcutaneous capsaicin increased blood pressure by 9%, had no effect on renal hemodynamics, but decreased urinary flow and sodium excretion by 30% to 40%. These antidiuretic and antinatriuretic, but not blood pressure, responses were absent after intracerebroventricular losartan administration to block angiotensin II type 1 receptors. By contrast, in the SHRSP, although subcutaneous capsaicin raised blood pressure and renal blood flow, neither glomerular filtration rate, urinary flow, nor sodium excretion changed, and this pattern of responses was unaffected after intracerebroventricular losartan. However, an intracerebroventricular infusion of angiotensin II increased basal blood pressure and fluid output, and the capsaicin challenge elicited vasopressor, antidiuretic, and antinatriuretic responses similar in magnitude to those observed in the Wistar rats. The capsaicin challenge in the SHRSP also caused a slowly developing, long-lasting fall in blood pressure and fluid excretion. These findings show that angiotensin II is a necessary component in the somatorenal reflex in normotensive rats but that endogenous angiotensin II is unable to exert this role in SHRSP. PMID- 11408380 TI - Neural mechanisms subserving central angiotensinergic influences on plasma renin in sheep. AB - The mechanisms and brain regions subserving the suppression of plasma renin concentration caused by intracerebroventricular (ICV) infusion of angiotensin II were studied in sodium-depleted sheep. Infusion of angiotensin II (3 microg/h for 1 hour) into the lateral ventricle reduced plasma renin from 4.3+/-0.4 to 1.6+/ 0.2 pmol angiotensin I/mL per hour at 1 hour after the commencement of infusion. This change persisted for at least another 90 minutes and was blocked by concomitant ICV infusion of the AT(1) antagonist losartan (1 mg/h). Arterial pressure did not change, but plasma vasopressin secretion was increased. ICV infusion of losartan (1 mg/h) significantly increased plasma renin in sodium depleted sheep. The reduction of plasma renin concentration in response to either ICV angiotensin II or hypertonic NaCl (0.75 mol/L at 1 mL/h) and the increase in response to ICV losartan was prevented in sheep in which the lamina terminalis of the brain had been ablated. Lesions in the median eminence (MEL), which blocked the increased plasma vasopressin levels, did not prevent suppression of plasma renin in response to ICV angiotensin II. However, bilateral renal denervation largely blocked this inhibition of plasma renin concentration but not the increased plasma renin resulting from ICV infusion of losartan in sodium-depleted sheep. The results show that AT(1) receptors, probably located in the lamina terminalis, mediate a central inhibitory influence of angiotensin II on renin secretion. This inhibition of renin release is probably due to a reduction in activity of renal sympathetic nerves innervating the juxtaglomerular apparatus of the kidney. PMID- 11408381 TI - Central orexin-A augments sympathoadrenal outflow in conscious rabbits. AB - We determined the cardiovascular and neurohormonal responses to intracerebroventricular administration of orexin-A in conscious rabbits. Intracerebroventricular injection of orexin-A elicited dose-related increases in mean arterial pressure and renal sympathetic nerve activity. Peak values of mean arterial pressure and renal sympathetic nerve activity induced by intracerebroventricular injection of 100 pmol of orexin-A (14.0+/-0.7 mm Hg and 55.4+/-14.9%, respectively) were obtained at 40 and 25 minutes after injection, respectively. Plasma epinephrine and glucose concentrations were significantly increased at 60 and 90 minutes after intracerebroventricular injection of orexin A (control versus 90 minutes; for epinephrine, 38.0+/-12.8 versus 167.5+/-42.5 pg/mL, P<0.01; for glucose, 6.66+/-0.18 versus 7.75+/-0.14 mmol/L, P<0.01). Plasma norepinephrine and insulin concentrations increased at 60 and 90 minutes but did not attain significant values. Intracerebroventricular injection of orexin-A also caused significant increases in plasma vasopressin concentrations. However, pretreatment with an intravenous injection of pentolinium (5 mg/kg), a ganglion-blocking agent, abolished these cardiovascular and neurohormonal responses. On the other hand, intravenous injection of the same dose of orexin-A (100 pmol) used in the intracerebroventricular experiment failed to cause any cardiovascular and renal sympathetic nerve responses. These results suggest that intracerebroventricular orexin-A acts in the central nervous system and activates sympathoadrenal outflow, resulting in increases in arterial pressure and plasma glucose levels in conscious rabbits. PMID- 11408382 TI - Effects of an angiotensin-converting enzyme inhibitor and a beta-blocker on cerebral arteriolar dilatation in hypertensive rats. AB - We examined the effects of the angiotensin-converting enzyme inhibitor perindopril and the beta-blocker propranolol on dilator responses of cerebral arterioles in chronic hypertension. Dilator responses to acute hypotension were examined in untreated Wistar-Kyoto rats (WKY) and stroke-prone spontaneously hypertensive rats (SHRSP) that were untreated or treated for 3 months with a low (0.3 mg. kg(-1). day(-1)) or a high (2 mg. kg(-1). day(-1)) dose of perindopril or a dose of propranolol (250 mg. kg(-1). day(-1)) alone or in combination with the low dose of perindopril. Pressure (servo-null) and diameter were measured in cerebral arterioles during acute reductions in arterial pressure both before and during maximal dilatation (EDTA). The high dose of perindopril or the combination of propranolol and perindopril normalized cerebral arteriolar pressure (52+/-2 [mean+/-SEM], 49+/-2 mm Hg versus 50+/-2 mm Hg in WKY and 96+/-3 mm Hg in untreated SHRSP; P<0.05). In contrast, the low dose of perindopril or propranolol alone did not normalize arteriolar pressure (74+/-2 mm Hg and 58+/-3 mm Hg). Both the low and high doses of perindopril improved autoregulatory dilatation, maximal dilatation, and dilator reserve of cerebral arterioles in SHRSP, with the low dose of perindopril being almost as effective as the high dose of perindopril. Propranolol alone did not significantly improve dilator function of cerebral arterioles. Furthermore, dilator function of cerebral arterioles was not further improved by the addition of propranolol to the low dose of perindopril. These findings suggest that angiotensin-converting enzyme inhibitors, such as perindopril, may be more effective than propranolol in attenuating the impairment of cerebral autoregulatory vasodilatation, maximal dilatation, and dilator reserve during treatment of chronic hypertension. PMID- 11408383 TI - Apoptosis is not increased in myocardium overexpressing type 2 angiotensin II receptor in transgenic mice. AB - To determine whether angiotensin type 2 (AT(2)) receptor stimulation induces apoptosis in cardiomyocytes in vivo, we developed transgenic mice overexpressing the AT(2) receptor in a cardiac-specific manner, using the alpha-myosin heavy chain promoter. Ten- to 12-week-old male homozygous transgenic mice (n=44) and wild-type mice (n=44) were used. Both transgenic and wild-type mice were given either saline (control), a subpressor dose of angiotensin II (100 ng. kg(-1). min(-1)), a pressor dose of angiotensin II (1000 ng. kg(-1). min(-1)) for 14 days, a pressor dose of angiotensin II for 28 days to investigate the effects of stimulation on both angiotensin type 1 (AT(1)) and AT(2) receptors, the AT(1) antagonist L158809 alone, or a combination of angiotensin II (1000 ng. kg(-1). min(-1)) and L158809 for 14 days to investigate the effects of selective AT(2) receptor stimulation. Apoptosis was analyzed in paraffin-embedded ventricular sections by the terminal deoxynucleotidyl-transferase-mediated dUTP nick-end labeling (TUNEL) technique. In both transgenic and wild-type mice, administration of a subpressor dose of angiotensin II, L158809, or a combination of angiotensin II and L158809 did not significantly affect the tail-cuff blood pressure or heart to-body weight ratio, whereas administration of a pressor dose of angiotensin II for 14 or 28 days significantly increased blood pressure and the heart-to-body weight ratio. However, there was no statistical difference between the effects of angiotensin II in transgenic and wild-type mice. The number of TUNEL-positive nuclei was approximately 0 to 10 per 100 000 cardiomyocytes, with no difference between transgenic and wild-type mice, regardless of saline infusion or any stimulation. In infarcted canine myocardial tissue sections for positive control, the number of TUNEL-positive nuclei was increased by 13.8 to 19.1 times compared with those in the noninfarcted myocardium. In conclusion, angiotensin II infusion for a period of 28 days failed to induce cardiomyocyte apoptosis regardless of the presence or absence of cardiac AT(2) receptor overexpression. It is unlikely that in mice the AT(2) receptor is a strong signal to induce cardiomyocyte apoptosis in vivo. PMID- 11408384 TI - Coronary hemodynamic and ventricular responses to angiotensin type 1 receptor inhibition in SHR: interaction with angiotensin type 2 receptors. AB - This study was designed to determine the effects of angiotensin II type 1 (AT(1)) receptor inhibition on coronary hemodynamics and ventricular mass and hydroxyproline content and the additive effects of angiotensin II type 2 (AT(2)) receptor inhibition in spontaneously hypertensive rats (SHR). The selective AT(1) receptor antagonist candesartan (10 mg/kg per day) was administered alone or in combination with the AT(2) receptor antagonist PD 123319 (50 mg/kg per day) for 12 weeks. Control SHR received placebo for the same period. Left and right ventricular coronary blood flow, blood flow reserve, and minimal coronary vascular resistance were determined by using radiomicrospheres in male 35-week old rats. Mean arterial pressure; total peripheral resistance; left and right ventricular, renal, and aortic weights; and hydroxyproline concentration were also determined. Candesartan reduced mean arterial pressure and left ventricular, renal, and aortic masses, as well as hydroxyproline concentration and minimal coronary vascular resistance of both ventricles. PD 123319 partially prevented the hypotensive effect of AT(1) receptor inhibition and reversed the effect on myocardial hydroxyproline concentration. These data suggest that AT(2) receptors contribute to the hypotensive and antifibrotic effects but not the coronary hemodynamic improvement or reduced left ventricular mass of AT(1) receptor inhibition in these adult SHR. PMID- 11408385 TI - Relation of age to left ventricular function and systemic hemodynamics in uncomplicated mild hypertension. AB - Previous studies in normotensive subjects have shown a slight decline in resting left ventricular pump function and midwall contractility with aging. We examined the relations of age to these variables and to peripheral resistance and vascular stiffness in 272 asymptomatic, unmedicated adults (25 to 80 years old) who had uncomplicated essential hypertension. Cardiac and carotid ultrasound and carotid pressure waveforms were obtained to measure left ventricular dimensions, endocardial and midwall left ventricular shortening, stroke index and cardiac index, end-systolic stress, and pulse pressure/stroke index and beta, pressure dependent and independent measures of vascular stiffness, respectively. Endocardial and midwall stress-corrected left ventricular shortening assessed ventricular performance. Cardiac index and TPRI did not change with age in either gender, with age-related increases in systolic pressure offset by increasingly concentric ventricular geometry in women and enhanced ventricular systolic function in men. In contrast to the lack of age-related change in traditional hemodynamic indexes, pulse pressure/stroke volume and beta strongly increased with age (P<0.001). Thus, in uncomplicated, relatively mild essential hypertension, neither cardiac index nor peripheral resistance is associated with age. This hemodynamic stability is associated with age-related increased concentricity of ventricular geometry in women and increased ventricular performance indexes in hypertensive men. Vascular stiffness progressively increases with age, independent of change in mean pressure or resistance, possibly contributing to increased rates of cardiovascular events in older individuals. PMID- 11408386 TI - Calcium channel blocker nifedipine slows down progression of coronary calcification in hypertensive patients compared with diuretics. AB - Calcium controls numerous events within the vessel wall. Permeability of the endothelium is calcium dependent, as are platelet activation and adhesion, vascular smooth muscle proliferation and migration, and synthesis of fibrous connective tissue. Double-helix computerized tomography is a noninvasive technique that can detect, measure, and compare coronary calcification in the coronary arteries. Using this method, our objective was to determine whether administration of nifedipine once daily in lieu of diuretics in high-risk hypertensive patients will arrest or slow down the progression of coronary artery calcification. The study was designed as a side arm of INSIGHT (International Nifedipine Study: Intervention as Goal for Hypertension Therapy), aimed to show the efficacy of nifedipine once daily versus co-amilozide (hydrochlorothiazide 25 mg, amiloride 2.5 mg) in high-risk hypertensive patients. A total of 201 patients with a total calcium score of >/=10 at the onset of study who underwent an annual double-helix computerized tomography for 3 years were analyzed for efficacy. Inhibition of coronary calcium progression was significant in the nifedipine versus the co-amilozide group during the first year (3.18% versus 27%, respectively, P=0.02), not significant during the second year (28.5% versus 47%, respectively, P=0.14), and significant during the third year (40% versus 78%, respectively, P=0.02). The results point to a slower progression of coronary calcification in hypertensive patients on nifedipine once daily versus co amilozide. PMID- 11408387 TI - Arterial calcification and calcium antagonists: what does it mean? PMID- 11408388 TI - Polymorphisms in the hANP (human atrial natriuretic peptide) gene, albuminuria, and hypertension. AB - Atrial natriuretic peptide (ANP) jointly affects kidney function and blood pressure homeostasis and is a candidate susceptibility gene for both essential hypertension and kidney disease. We evaluated the relation between the ScaI and BstXI polymorphisms of the human ANP (hANP) gene, hypertension, and albuminuria in a clinical cohort of 1033 subjects, including type 1 and type 2 diabetic patients, nondiabetic subjects with essential hypertension, and nondiabetic normotensive control subjects. Microalbuminuria was present in 15%, 29%, and 2%, respectively, of type 1 diabetic, type 2 diabetic, and nondiabetic patients. Macroalbuminuria was present in 9% of type 1 diabetics, 21% of type 2 diabetics, and 31% of nondiabetics. Prevalence of hypertension was 31%, 58%, and 61% in normoalbuminuric, microalbuminuric, and macroalbuminuric subjects, respectively (P<0.0001). Genotype distributions were in Hardy-Weinberg equilibrium in all 4 patient subgroups. The frequency of the ScaI mutated allele (A(1)) was significantly lower in hypertensive than in control subjects (11% versus 19%, P=0.018) and in patients with macroalbuminuria (5%) as compared with normoalbuminuric subjects (16%; P<0.0001). In a nominal logistic model adjusting for gender, age, obesity, diabetes, micro/macroalbuminuria, and hypertension, the A(1) allele was independently associated with macroalbuminuria (odds ratio, 0.57; confidence interval, 1.39 to 3.59; P=0.003) but not with hypertension. In the same model, the frequency of the BstXI mutated allele (T(708)) was increased in the presence of microalbuminuria (odds ratio, 2.25; confidence interval, 1.39 to 3.59; P<0.001). We conclude that the mutated genotypes of the ScaI polymorphism are negatively associated with overt nephropathy, whereas the mutated genotypes of BstXI polymorphism are positively associated with microalbuminuria. hANP gene variants may exert a protective effect against the development and progression of kidney damage in diabetes. PMID- 11408389 TI - Expression and distribution of NOS1 and NOS3 in the myocardium of angiotensin II infused rats. AB - Studies have indicated a complex functional interaction between angiotensin (Ang) II and NO in the heart. The purpose of the present study was to examine the protein expression and tissue distribution of NO synthases 1 (NOS1) and 3 (NOS3) in the myocardium of rats that underwent continuous infusion of Ang II at 2 different rates (10 and 40 ng. kg(-1). min(-1)) for 6 days. Mean arterial pressure increased by approximately 15 mm Hg in rats infused with Ang II at 40 ng. kg(-1). min(-1), but it remained close to the values observed in saline infused rats ( approximately 110 mm Hg) when Ang II was infused at 10 ng. kg(-1). min(-1). The protein expression of a 160-kDa NOS1 and a 135-kDa NOS3 were found to increase ( approximately 200%) in the myocardium of rats infused with both subpressor and pressor doses of Ang II. Immunohistochemistry studies showed that NOS1 and NOS3 are differentially expressed in myocardial cells. NOS1 was detected in cardiac myocytes and in smooth muscle cells of small and large coronary arteries, whereas NOS3 was detected in the endothelium and in perivascular and interstitial tissues, but NOS3 was not detected in cardiac or smooth muscle cells. Ang II infusion enhanced the tissue immunoreactivity of both isoforms in their specific locations but did not change the distribution throughout the myocardium. Myocardium staining with anti-angiotensin type 1 (AT(1)) receptor antibody indicated that AT(1) receptor is expressed in cardiac myocytes, coronary smooth muscle cells, and interstitial and perivascular tissues. Ang II infusion did not change the protein expression and distribution of AT(1) receptor in the myocardium. These results indicate that long-term increases in the circulating levels of Ang II modulate the protein expression of NOS1 and NOS3 and, consequently, the function of the local myocardial NO system. PMID- 11408390 TI - Vasoactive drugs influence aortic augmentation index independently of pulse-wave velocity in healthy men. AB - Aortic augmentation index, a measure of central systolic blood pressure augmentation arising mainly from pressure-wave reflection, increases with vascular aging. The augmentation index is influenced by aortic pulse-wave velocity (related to aortic stiffness) and by the site and extent of wave reflection. To clarify the relative influence of pulse-wave velocity and wave reflection on the augmentation index, we studied the association between augmentation index, pulse-wave velocity, and age and examined the effects of vasoactive drugs to determine whether altering vascular tone has differential effects on pulse-wave velocity and the augmentation index. We made simultaneous measurements of the augmentation index and carotid-to-femoral pulse-wave velocity in 50 asymptomatic men aged 19 to 74 years at baseline and, in a subset, during the administration of nitroglycerin, angiotensin II, and saline vehicle. The aortic augmentation index was obtained by radial tonometry (Sphygmocor device, PWV Medical) with the use of an inbuilt radial to aortic transfer function. In multiple regression analysis, the aortic augmentation index was independently correlated only with age (R=0.58, P<0.0001). Nitroglycerin (3 to 300 microg/min IV) reduced the aortic augmentation index from 4.8+/-2.3% to -11.9+/-5.3% (n=10, P<0.002). Angiotensin II (75 to 300 ng/min IV) increased the aortic augmentation index from 9.3+/-2.4% to 18.3+/-2.9% (n=12, P<0.001). These drugs had small effects on aortic pulse-wave velocity, producing mean changes from baseline of <1 m/s (each P<0.05). In healthy men, vasoactive drugs may change aortic augmentation index independently from aortic pulse-wave velocity. PMID- 11408391 TI - Peripheral "oscillatory" compliance is associated with aortic augmentation index. AB - The augmentation index (AIx) and "oscillatory" compliance (C(2)) are wave contour analysis parameters for the central aorta (P(ao)) and radial artery pressure wave (P(rad)), respectively. Both are sensitive to cardiovascular risk factors such as aging, hypertension, and diabetes and have been proposed as prognostic markers for cardiovascular disease. In this work, we studied the relation between both. We first calculated P(rad) corresponding to a typical aortic A-type (AIx >0.15) and C-type wave (AIx <0), taken from the literature, by using a generalized aorta radial pressure transfer function. P(rad) corresponding to C-type waves yielded the highest C(2) value. We further used simultaneously measured aortic and radial artery pressure in 45 human subjects age 34 to 84 years (63+/-12 [SD]) at baseline and after administration of nitroglycerin to calculate AIx(meas) and C(2), respectively. Transfer function was used to calculate reconstructed aortic pressure and AIx(rec). AIx(rec) underestimates AIx(meas) by 0.03+/-0.16, but both values correlate well (r=0.64; P<0.001). C(2) and AIx were inversely correlated (r=-0.36; P<0.001 for AIx(meas); r=-0.30; P<0.01 for AIx(rec)). Both AIx(meas) (0.06+/-0.17 versus 0.20+/-0.21; P<0.01) and AIx(rec) (0.04+/-0.12 versus 0.16+/ 0.16; P<0.001) were lower after nitroglycerin, whereas C(2) increased only nonsignificantly (0.080+/-0.036 versus 0.071+/-0.042). C(2) is related to AIx and reflects, at least in part, hemodynamic changes affecting central aortic pressure. Nevertheless, given the model assumptions and computational steps associated with calculating C(2), AIx could be a more appropriate parameter to use in the clinical setting because it is determined directly from the pressure wave contour. PMID- 11408392 TI - Captopril suppression versus salt loading in confirming primary aldosteronism. AB - This prospective study was designed to compare the captopril suppression test with the salt-loading approach to confirm the diagnosis of primary aldosteronism. A total of 49 patients were referred with a presumed diagnosis of primary aldosteronism. The captopril test was performed in the morning with patients in the seated position after overnight fasting. Blood samples for plasma aldosterone were obtained before captopril administration (25 mg PO) and again 2 hours later. Patients were then subjected to a high salt diet (300 mmol sodium per day for 3 days). On the third day, urinary sodium (24 hours) was measured, and plasma aldosterone levels were measured at 8:00 AM (recumbent) and at noon (standing). Of the 49 patients, 44 had nonsuppressible aldosterone concentrations with all the clinical characteristics of primary aldosteronism: 22 patients had surgically confirmed unique adenoma, and 22 patients had presumed bilateral hyperplasia. There was a significant correlation between plasma aldosterone values of salt loaded patients (mean of 8:00 AM and noon results) and the values 2 hours after captopril administration (r=0.8, P<0.01). Plasma aldosterone cumulative distribution curves in primary aldosteronism patients (adenoma and hyperplasia) were not significantly different between the 2 suppression tests. Our results showed that the captopril suppression test is as effective as sodium loading in confirming the diagnosis of primary aldosteronism. PMID- 11408393 TI - ET(A) receptor antagonist prevents blood pressure elevation and vascular remodeling in aldosterone-infused rats. AB - Increased endothelin-1 may be associated with elevation of blood pressure (BP) and promotion of vascular hypertrophy, especially in salt-sensitive hypertension. Mineralocorticoid hypertension has been associated with activation of the endothelin system. We evaluated whether in aldosterone-infused rats the selective endothelin type A receptor-antagonist BMS 182874 prevents BP elevation and vascular hypertrophy. Rats were infused with aldosterone (0.75 microg/h) subcutaneously via a mini-osmotic pump and were offered 1% NaCl in the drinking water+/-BMS 182874 (40 mg/kg in food) for 6 weeks. Systolic BP was monitored by the tail-cuff method, and vascular changes of mesenteric arteries were evaluated using a pressurized myograph. Aldosterone-infusion significantly increased BP to 151+/-7 mm Hg compared with controls (108+/-4 mm Hg, P<0.01). BMS 182874 normalized BP (117+/-4 mm Hg). Media cross-sectional area of aorta was significantly increased by aldosterone infusion (P<0.05), and BMS treatment normalized it (P<0.001). Aldosterone infusion increased media width and media-to lumen ratio of mesenteric resistance arteries (17.6+/-0.4 microm and 7.5+/-0.4%) compared with controls (14.2+/-0.5 microm, P<0.01, and 5.9+/-0.1%, P<0.05). BMS 182874 normalized media and media-to-lumen ratio (15.1+/-0.6 microm and 5.7+/ 0.1%, both P<0.01). In conclusion, the endothelin type A receptor antagonist attenuated BP elevation and prevented vascular remodeling or hypertrophy of aorta and mesenteric resistance arteries in aldosterone-infused rats. These results suggest a role for endothelin-1 in BP elevation and structural alterations of large and small vessels in aldosterone and salt-induced hypertension. PMID- 11408394 TI - HMG-CoA reductase inhibitors improve endothelial dysfunction in normocholesterolemic hypertension via reduced production of reactive oxygen species. AB - 3-Hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors (statins) significantly reduce cardiovascular mortality associated with hypercholesterolemia. There is evidence that statins exert beneficial effects in part through direct effects on vascular cells independent of lowering plasma cholesterol. We characterized the effect of a 30-day treatment with atorvastatin in normocholesterolemic, spontaneously hypertensive rats (SHR). Systolic blood pressure was significantly decreased in atorvastatin-treated rats (184+/-5 versus 204+/-6 mm Hg for control). Statin therapy improved endothelial dysfunction, as assessed by carbachol-induced vasorelaxation in aortic segments, and profoundly reduced angiotensin II-induced vasoconstriction. Angiotensin type 1 (AT(1)) receptor, endothelial cell NO synthase (ecNOS), and p22phox mRNA expression were determined with quantitative reverse transcription-polymerase chain reaction. Atorvastatin treatment downregulated aortic AT(1) receptor mRNA expression to 44+/-12% of control and reduced mRNA expression of the essential NAD(P)H oxidase subunit p22phox to 63+/-7% of control. Aortic AT(1) receptor protein expression was consistently decreased. Vascular production of reactive oxygen species was reduced to 62+/-12% of control in statin-treated SHR, as measured with lucigenin chemiluminescence assays. Accordingly, treatment of SHR with the AT(1) receptor antagonist fonsartan improved endothelial dysfunction and reduced vascular free radical release. Moreover, atorvastatin caused an upregulation of ecNOS mRNA expression (138+/-7% of control) and an enhanced ecNOS activity in the vessel wall (209+/-46% of control). Treatment of SHR with atorvastatin causes a significant reduction of systolic blood pressure and a profound improvement of endothelial dysfunction mediated by a reduction of free radical release in the vasculature. The underlying mechanism could in part be based on the statin induced downregulation of AT(1) receptor expression and decreased expression of the NAD(P)H oxidase subunit p22phox, because AT(1) receptor activation plays a pivotal role for the induction of this redox system in the vessel wall. PMID- 11408395 TI - Mutations in the Na-Cl cotransporter reduce blood pressure in humans. AB - The relationship between salt homeostasis and blood pressure has remained difficult to establish from epidemiological studies of the general population. Recently, mendelian forms of hypertension have demonstrated that mutations that increase renal salt balance lead to higher blood pressure, suggesting that mutations that decrease the net salt balance might have the converse effect. Gitelman's syndrome, caused by loss of function mutations in the Na-Cl cotransporter of the distal convoluted tubule (NCCT), features inherited hypokalemic alkalosis with so-called "normal" blood pressure. We hypothesized that the mild salt wasting of Gitelman's syndrome results in reduced blood pressure and protection from hypertension. We have formally addressed this question through the study of 199 members of a large Amish kindred with Gitelman's syndrome. Through genetic testing, family members were identified as inheriting 0 (n=60), 1 (n=113), or 2 (n=26) mutations in NCCT, permitting an unbiased assessment of the clinical consequences of inheriting these mutations by comparison of the phenotypes of relatives with contrasting genotypes. The results demonstrate high penetrance of hypokalemic alkalosis, hypomagnesemia, and hypocalciuria in patients inheriting 2 mutant NCCT alleles. In addition, the NCCT genotype was a significant predictor of blood pressure, with homozygous mutant family members having significantly lower age- and gender-adjusted systolic and diastolic blood pressures than those of their wild-type relatives. Moreover, both homozygote and heterozygote subjects had significantly higher 24-hour urinary Na(+) than did wild-type subjects, reflecting a self-selected higher salt intake. Finally, heterozygous children, but not adults, had significantly lower blood pressures than those of the wild-type relatives. These findings provide formal demonstration that inherited mutations that impair renal salt handling lower blood pressure in humans. PMID- 11408396 TI - Effect of extracellular matrix elements on angiotensin II-induced calcium release in vascular smooth muscle cells from normotensive and hypertensive rats. AB - The interaction of the vascular smooth muscle cells (VSMCs) with the components of the matrix determines several functions of the cell, such as growth and differentiation. In contrast, an alteration in angiotensin (Ang) II-induced Ca(2+) mechanisms in VSMCs was reported in genetic hypertension. In this study, we wished to assess the effect of different components of the extracellular matrix on the increase of [Ca(2+)](i) induced by Ang II in VSMCs from spontaneously hypertensive rats (SHR) compared with those from normotensive Wistar-Kyoto rats (WKY). Results demonstrate for the first time that elements of the extracellular matrix modulate the Ang II-induced Ca(2+) transport mechanisms. This modulation is different in cells from WKY compared with those from SHR. Thus, growing cells from SHR on collagen I, collagen IV, fibronectin, vitronectin, or Matrigel induced a significant decrease in Ang II-induced Ca(2+) release from internal stores, whereas in cells from WKY, no effect could be observed except for those grown on collagen I, which increased Ca(2+) release. Fibronectin and vitronectin, however, induced a decrease in Ang II-induced Ca(2+) influx in WKY, whereas no effect could be observed in SHR. Conversely, collagen I and collagen IV induced an increase in this influx in SHR but not in WKY, whereas Matrigel increased the influx in both strains. These results suggest a modulation of the Ang II-associated signaling events by the matrix elements via the focal adhesion points. The understanding of these synergies should provide insight into issues such as development of hypertrophy of large vessels in hypertension. PMID- 11408397 TI - Normal blood pressure and renal function in mice lacking the bradykinin B(2) receptor. AB - Telemetric blood pressure determinations, heart rate measurements, and pressure natriuresis-diuresis experiments were used to characterize cardiovascular and renal function in bradykinin B(2) receptor knockout mice fed mouse chow containing 0.25% NaCl or mouse chow containing 4% NaCl. In B(2) receptor knockout mice fed usual mouse chow, the mean arterial blood pressure leveled between 108+/ 1 and 110+/-3 mm Hg, and the heart rate leveled between 520+/-26 and 525+/-29 bpm, values that were not different from those measured in B(1) receptor knockout mice or 129Sv/J control mice. Increasing dietary salt intake did not affect mean arterial blood pressure and heart rate. Accordingly, pressure-natriuresis curves, pressure-diuresis curves, renal blood flow, and glomerular filtration rate were not different between B(2) receptor knockout and 129Sv/J mice. Increasing dietary salt intake to 4% increased renal blood flow to levels between 8.41 and 9.50 mL/min per gram kidney wet weight in 129Sv/J mice, whereas in B(2) receptor deficient mice, renal blood flow was not affected and ranged between 6.85 and 7.88 mL/min per gram kidney wet weight. Other renal function parameters were not affected. Absence of B(2) receptor function was verified in B(2) receptor knockout mice with bradykinin infusion. These data suggest that the absence of B(2) receptor function does not necessarily make B(2) receptor knockout mice hypertensive or induce salt sensitivity. Presumably, differences in the genetic background or an adaptation to the loss of B(2) receptor function may account for these results, in contrast with earlier reports involving B(2) receptor knockout mice. We hold the latter possibility to be more likely and to be a fruitful possibility for future research. PMID- 11408398 TI - Impaired insulin-like growth factor I vasorelaxant effects in hypertension. AB - Insulin-like growth factor I (IGF-I) can be considered a factor potentially involved in arterial hypertension not only for its growth-promoting features but also for its effects on vascular tone. Nevertheless, the actions of the hormone on vascular reactivity are still unexplored in hypertension. Therefore, the vasodilation induced by increasing doses of IGF-I and the modulation of norepinephrine vasoconstriction induced by low levels of the hormone were tested on aortic rings of spontaneously hypertensive and normotensive rats. The results indicate that the vasodilation evoked by IGF-I is impaired in hypertensive rats (Delta% of maximal vasorelaxation, 30+/-1 versus 41+/-1; P<0.01), and after the removal of endothelium or the inhibition of endothelial NO synthase, the vasodilation evoked by the hormone was blunted in both rat strains and became similar between hypertensive and normotensive rats (Delta% of maximal vasorelaxation, 21+/-1 versus 20+/-1; P=NS). Moreover, IGF-I does not show any effect on norepinephrine vasoconstriction in hypertensive rats, and this alteration may depend on the lack of sensitizing effect exerted by IGF-I on alpha(2)-adrenergic-evoked NO vasorelaxation. The defect in IGF-I vascular action is also present in young spontaneously hypertensive rats (age 5 weeks). In conclusion, our data demonstrate that IGF-I vasorelaxant properties are impaired in spontaneously hypertensive rats, suggesting that such defect may play a causative or permissive role in the development of hypertensive conditions. PMID- 11408399 TI - Effect of insulin and angiotensin II on cell calcium in human skin fibroblasts. AB - We have recently shown that insulin attenuates angiotensin II-induced intracellular Ca(2+) mobilization in human skin fibroblasts from normotensive subjects. This study was designed to investigate the effects of angiotensin II and the interactions between insulin and angiotensin II on intracellular Ca(2+) mobilization in skin fibroblasts from patients with essential hypertension. Fibroblasts were obtained from 9 normotensives and 18 hypertensives. Spectrofluorophotometric free Ca(2+) measurement was performed in monolayers of 24-hour serum-deprived cells. Resting intracellular Ca(2+) level and angiotensin II-stimulated intracellular Ca(2+) peak were higher in fibroblasts from hypertensives compared with those from normotensives. The effect of acute insulin exposure was evaluated in fibroblasts from hypertensives subdivided on the basis of insulin sensitivity. In insulin-sensitive hypertensives, insulin significantly blunted the effects of angiotensin II on intracellular Ca(2+) response, whereas in insulin-resistant patients, insulin did not modify intracellular Ca(2+) response to angiotensin II. Pertussis toxin, a G(ialpha)-inhibitor, reduced angiotensin II-stimulated Ca(2+) peak in insulin-sensitive but not in insulin resistant hypertensives. In conclusion, the effects of angiotensin II on intracellular Ca(2+) mobilization are more pronounced in fibroblasts from hypertensives compared with those from normotensives, and the inhibitory effect of insulin is blunted in insulin-resistant hypertensives by a G(ialpha) pertussis toxin-sensitive abnormality. PMID- 11408400 TI - A defect in glycogen synthesis characterizes insulin resistance in hypertensive patients with type 2 diabetes. AB - A subgroup of patients with type 2 diabetes shows a clustering of abnormalities such as peripheral insulin resistance, hypertension, and microalbuminuria. To evaluate whether these traits reflect intrinsic disorders of cell function rather than in vivo environmental effects, we studied a group of 7 nondiabetic hypertensive subjects with an altered albumin excretion rate (AER) (HyMA+) and 3 groups of patients with type 2 diabetes: 7 with normal blood pressure and normal AER (DH-MA-), 7 with high blood pressure and normal AER (DH+MA-), and 7 with both high blood pressure and altered AER (DH+MA+). Glucose disposal was measured during an hyperinsulinemic clamp (40 mU. m(2)(-1). min(-1)) with primed deuterated [6.6 (2)H(2)] glucose infusion. In the same subjects, a skin biopsy was performed and the following parameters were investigated: glucose transport (as determined by [(3)H]2-deoxyglucose uptake); glycogen synthase activity (as determined by [(14)C] glucose incorporation from UDP-[U-(14)C] glucose into glycogen); glycogen phosphorylase activity (as measured by the incorporation of [U-(14)C]glucose 1-phosphate into glycogen); and total glycogen content. In vivo glucose disposal was significantly reduced in DH+MA- and DH+MA+, with respect to DH-MA-, HyMA+, and controls. Insulin-stimulated glucose transport was similar in the 3 groups of patients with diabetes. A significant reduction of intracellular glycogen content was observed in DH+MA- and DH+MA+ compared with DH-MA- in both basal and insulin-stimulated conditions, probably because of a major impairment of glycogen synthase activity. Glycogen phosphorylase activity did not show differences between the groups. These results suggest that (1) the combination of type 2 diabetes with hypertension and altered AER is associated with impaired insulin sensitivity, and (2) intrinsic, possibly genetic, factors may account for increased peripheral insulin resistance in hypertensive microalbuminuric patients with type 2 diabetes, pointing to the reduction of glycogen synthase activity as a shared common defect. PMID- 11408401 TI - A comparison between systolic and diastolic pulse contour analysis in the evaluation of arterial stiffness. AB - Several methodologically independent measures of arterial stiffness derived from either the systolic or diastolic segments of the arterial pulse have been proposed. The exact nature of the large and small artery elasticity indices (C1 and C2, respectively) derived from diastolic pulse contour analysis remains largely unexplored, although C2 has controversially been termed to be "oscillatory" and "reflective." We investigated the relation between C2 and, respectively, a prototype of arterial reflectivity (ie, the augmentation index, AIx) and a covariate of arterial reflectivity (body height). A validated transfer function is used to transform a tonometrically obtained radial pressure wave into an ascending aortic pressure wave, from which AIx is derived using systolic pulse contour analysis. Diastolic pulse contour analysis using a modified Windkessel model is used to derive C1 and C2. One hundred subjects, who were free from atherothrombotic disease and 19 to 77 years of age, with a wide pressure range (97 to 186/52 to 104 mm Hg) were studied. Mean values of C1, C2, AIx, and body height were, respectively, 13.8+/-4.3 mL/mm Hgx10, 5.9+/-3.1 mL/mm Hgx100, 128.5+/-24.9%, and 169+/-9 cm. Coefficients of variation were 32.8% for C1, 33.3% for C2, and 6.7% for AIx. C2 was significantly and inversely correlated to AIx (r=-0.707, P<0.001). Both AIx and C2 were correlated to body height (r=-0.487, P<0.001, and r=0.514, P<0.001). In conclusion, the results of this study provide the first clinical evidence that validates a probable biophysical equivalent of the C2 element of a third-order, 4-element modified Windkessel model. We suggest that C2 is, at least in part, a measure of arterial wave reflectance. However, although short-term reproducibility of AIx is excellent, C2 showed markedly increased variability with the devices used. PMID- 11408403 TI - Normalization of blood pressure in a patient with severe orthostatic hypotension and supine hypertension using clonidine. PMID- 11408404 TI - Neurovascular contact of cranial nerve IX and X root-entry zone in hypertensive patients. PMID- 11408406 TI - Ventilation-perfusion inhomogeneity increases gas uptake: theoretical modeling of gas exchange. AB - Ventilation-perfusion (VA/Q) inhomogeneity was modeled to measure its effect on gas exchange in the presence of inspired mixtures of two soluble gases using a two-compartment computer model. Theoretical studies involving a mixture of hypothetical gases with equal solubility in blood showed that the effect of increasing inhomogeneity of distributions of either ventilation or blood flow is to paradoxically increase uptake of the gas with the lowest overall uptake in relation to its inspired concentration. This phenomenon is explained by the concentrating effects that uptake of soluble gases exert on each other in low VA/Q compartments. Repeating this analysis for inspired mixtures of 30% O(2) and 70% nitrous oxide (N(2)O) confirmed that, during "steady-state" N(2)O anesthesia, uptake of N(2)O is predicted to paradoxically increase in the presence of worsening VA/Q inhomogeneity. PMID- 11408407 TI - Ventilation-perfusion inhomogeneity increases gas uptake in anesthesia: computer modeling of gas exchange. AB - Ventilation-perfusion (VA/Q) inhomogeneity was modeled to measure its effect on overall gas exchange during maintenance-phase N(2)O anesthesia with an inspired O(2) concentration of 30%. A multialveolar compartment computer model was used based on physiological log normal distributions of VA/Q inhomogeneity. Increasing the log standard deviation of the distribution of perfusion from 0 to 1.75 paradoxically increased O(2) uptake (VO(2)) where a low mixed venous partial pressure of N(2)O [high N(2)O uptake (VN(2)O)] was specified. With rising mixed venous partial pressure of N(2)O, a threshold was observed where VO(2) began to fall, whereas VN(2)O began to rise with increasing VA/Q inhomogeneity. This phenomenon is a magnification of the concentrating effects that VO(2) and VN(2)O have on each other in low VA/Q compartments. During "steady-state" N(2)O anesthesia, VN(2)O is predicted to paradoxically increase in the presence of worsening VA/Q inhomogeneity. PMID- 11408408 TI - Effect of ventilation-perfusion inhomogeneity and N(2)O on oxygenation: physiological modeling of gas exchange. AB - Ventilation-perfusion (VA/Q) inhomogeneity was modeled to measure its effect on arterial oxygenation during maintenance-phase anesthesia involving an inspired mixture of 30% O(2) and either N(2)O or N(2). A multialveolar compartment computer model was constructed based on a log normal distribution of VA/Q inhomogeneity. Increasing the log SD of the distribution of blood flow from 0 to 1.75 produced a progressive fall in arterial PO(2) (Pa(O(2))). The fall was less steep in the presence of N(2)O than when N(2) was present instead. This was due mainly to the concentrating effect of N(2)O uptake on alveolar PO(2) in moderately low VA/Q compartments. The improvement in Pa(O(2)) when N(2)O was present instead of N(2) was greatest when the degree of VA/Q inhomogeneity was in the range typically seen in anesthetized patients. Models based on distributions of expired and inspired alveolar ventilation give quantitatively different results for Pa(O(2)). In the presence of VA/Q inhomogeneity, second-gas and concentrating effects may have clinically significant effects on arterial oxygenation even at "steady-state" levels of N(2)O uptake. PMID- 11408409 TI - Effects of isometric training on the elasticity of human tendon structures in vivo. AB - The present study aimed to investigate the effect of isometric training on the elasticity of human tendon structures. Eight subjects completed 12 wk (4 days/wk) of isometric training that consisted of unilateral knee extension at 70% of maximal voluntary contraction (MVC) for 20 s per set (4 sets/day). Before and after training, the elongation of the tendon structures in the vastus lateralis muscle was directly measured using ultrasonography while the subjects performed ramp isometric knee extension up to MVC. The relationship between the estimated muscle force and tendon elongation (L) was fitted to a linear regression, the slope of which was defined as stiffness of the tendon structures. The training increased significantly the volume (7.6+/-4.3%) and MVC torque (33.9+/-14.4%) of quadriceps femoris muscle. The L values at force production levels beyond 550 N were significantly shorter after training. The stiffness increased significantly from 67.5+/-21.3 to 106.2+/-33.4 N/mm. Furthermore, the training significantly increased the rate of torque development (35.8 +/- 20.4%) and decreased electromechanical delay (-18.4+/-3.8%). Thus the present results indicate that isometric training increases the stiffness and Young's modulus of human tendon structures as well as muscle strength and size. This change in the tendon structures would be assumed to be an advantage for increasing the rate of torque development and shortening the electromechanical delay. PMID- 11408410 TI - Physiological consequences of oxygen-dependent chloride binding to hemoglobin. AB - The PO(2)-dependent binding of chloride to Hb decreases the Cl(-) concentration of the red blood cell (RBC) intracellular fluid in venous blood to approximately 1-3 mmol/l less than that in arterial blood. This change is physiologically important because 1) Cl(-) is a negative heterotropic allosteric effector of Hb that competes for binding sites with 2,3-bisphosphoglycerate and CO(2) and decreases oxyhemoglobin affinity in several species; 2) it may help reconcile several longstanding problems with measured values of the Donnan ratios for Cl( ), HCO, and H(+) across the RBC membrane that are used to calculate total CO(2) carriage, ion flux rates, and membrane potentials; 3) it is a factor in the change in the dissociation constant for the combined nonvolatile weak acids of Hb associated with the Haldane effect; and 4) it diminishes the decrease in strong ion difference in the RBC intracellular fluid that would otherwise occur from the chloride shift and prevent the known increase of HCO concentration in that compartment. PMID- 11408411 TI - Gravity effects on regional lung ventilation determined by functional EIT during parabolic flights. AB - Gravity-dependent changes of regional lung function were studied during normogravity, hypergravity, and microgravity induced by parabolic flights. Seven healthy subjects were followed in the right lateral and supine postures during tidal breathing, forced vital capacity, and slow expiratory vital capacity maneuvers. Regional 1) lung ventilation, 2) lung volumes, and 3) lung emptying behavior were studied in a transverse thoracic plane by functional electrical impedance tomography (EIT). The results showed gravity-dependent changes of regional lung ventilation parameters. A significant effect of gravity on regional functional residual capacity with a rapid lung volume redistribution during the gravity transition phases was established. The most homogeneous functional residual capacity distribution was found at microgravity. During vital capacity and forced vital capacity in the right lateral posture, the decrease in lung volume on expiration was larger in the right lung region at all gravity phases. During tidal breathing, the differences in ventilation magnitudes between the right and left lung regions were not significant in either posture or gravity phase. A significant nonlinearity of lung emptying was determined at normogravity and hypergravity. The pattern of lung emptying was homogeneous during microgravity. PMID- 11408412 TI - Assessment of dry heat exchanges in newborns: influence of body position and clothing in SIDS. AB - A dramatic decrease of sudden infant death syndrome (SIDS) has been noted following the issuance of recommendations to adopt the supine sleeping position for infants. It has been suggested that the increased risk could be related to heat stress associated with body position. In the present study, the dry heat losses of small-for-gestational-age newborns nude or clothed were assessed and compared to see whether there is a difference in the ability to lose heat between the prone and supine positions. An anthropomorphic thermal mannequin was exposed to six environmental temperatures, ranging between 25 and 37 degrees C, in a single-walled, air-heated incubator. The magnitudes of heat losses did not significantly differ between the two body positions for the nude (supine 103.46 +/- 29.67 vs. prone 85.78 +/- 34.91 W/m(2)) and clothed mannequin (supine 59.35 +/- 21.51 vs. prone 63.17 +/- 23.06 W/m(2)). With regard to dry heat exchanges recorded under steady-state conditions, the results show that there is no association between body position and body overheating. PMID- 11408413 TI - Comparison of a space shuttle flight (STS-78) and bed rest on human muscle function. AB - The purpose of this investigation was to assess muscle fiber size, composition, and in vivo contractile characteristics of the calf muscle of four male crew members during a 17-day spaceflight (SF; Life and Microgravity Sciences Spacelab Shuttle Transport System-78 mission) and eight men during a 17-day bed rest (BR). The protocols and timelines of these two investigations were identical, therefore allowing for direct comparisons between SF and the BR. The subjects' age, height, and weight were 43 +/- 2 yr, 183 +/- 4 cm, and 86 +/- 3 kg for SF and 43 +/- 2 yr, 182 +/- 3 cm, and 82 +/- 4 kg for BR, respectively. Calf muscle strength was examined before SF and BR; on days 2, 8, and 12 during SF and BR; and on days 2 and 8 of recovery. Muscle biopsies were obtained before and within 3 h after SF (gastrocnemius and soleus) and BR (soleus) before reloading. Maximal isometric calf strength and the force-velocity characteristics were unchanged with SF or BR. Additionally, neither SF nor BR had any effect on fiber composition or fiber size of the calf muscles studied. In summary, no changes in calf muscle strength and morphology were observed after the 17-day SF and BR. Because muscle strength is lost during unloading, both during spaceflight and on the ground, these data suggest that the testing sequence employed during the SF and BR may have served as a resistance training countermeasure to attenuate whole muscle strength loss. PMID- 11408414 TI - Mechanical properties of alveolar epithelial cells in culture. AB - With the use of magnetic twisting cytometry, we characterized the mechanical properties of rat type II alveolar epithelial (ATII) cells in primary culture and examined whether the cells' state of differentiation and the application of deforming stresses influence their resistance to shape change. Cells were harvested from rat lungs as previously described (Dobbs LG. Am J Physiol Lung Cell Mol Physiol 258: L134-L147, 1990) and plated at a density of 1 x 10(6) cells/cm(2) in fibronectin-coated 96 Remova wells, and their mechanical properties were measured 2-9 days later. We show 1) that ATII cells form much stronger bonds with RGD-coated beads than they do with albumin- or acetylated low density lipoprotein-coated beads, 2) that RGD-mediated bonds seemingly "mature" during the first 60 min of bead contact, 3) that the apparent stiffness of ATII cells increases with days in culture, 4) that stiffness falls when the RGD-coated beads are intermittently oscillated at 0.3 Hz, and 5) that this fall cannot be attributed to exocytosis-related remodeling of the subcortical cytoskeleton. Although the mechanisms of force transfer between basement membrane, cytoskeleton, and plasma membrane of ATII cells remain to be resolved, such analyses undoubtedly require definition of the cell's mechanical properties. To our knowledge, the results presented here provide the first data on this topic. PMID- 11408415 TI - Comparison of OPS imaging and conventional capillary microscopy to study the human microcirculation. AB - Orthogonal polarization spectral (OPS) imaging is a new clinical technique for observation of the microcirculation of organ surfaces. For validation purposes, we compared OPS images of the nailfold skin with those obtained from conventional capillary microscopy at rest and during venous occlusion in 10 male volunteers. These images were computer analyzed to provide red blood cell velocity and capillary diameters of the same nailfold capillaries at rest and during venous occlusion. Results showed that OPS images provided similar values for red blood cell velocity and capillary diameter as those obtained from capillary microscopy images. OPS imaging, however, provided significantly better image quality, as shown by comparison of image contrast between OPS imaging and capillary microscopy. This made image analysis better and easier to perform. It is anticipated, therefore, that OPS imaging will become a new and powerful technique in the study of the human microcirculation in vivo because it can be used on human internal organs. PMID- 11408416 TI - Severe diabetes inhibits resistance exercise-induced increase in eukaryotic initiation factor 2B activity. AB - Rates of protein synthesis are reduced in severely diabetic rats. A potential mechanism through which insulin can stimulate protein synthesis is modulation of the activity of eukaryotic initiation factor 2B (eIF2B). The activity of this factor is elevated after exercise in nondiabetic rats but is markedly lower in skeletal muscle from nonexercised severely diabetic rats. We tested the hypothesis that a failure to increase eIF2B activity after exercise is one potential reason for a failure of severely diabetic rats to increase rates of protein synthesis after resistance exercise. Diabetic (partial pancreatectomy, plasma glucose >475 mg/dl) and nondiabetic male Sprague-Dawley rats (approximately 300 g) performed acute moderate-intensity resistance exercise or remained sedentary. Rates of protein synthesis were higher in nondiabetic rats and increased significantly with exercise, while no elevation was found in severely diabetic rats. The activity of eIF2B was higher (P < 0.05) in exercised nondiabetic than in sedentary nondiabetic rats (0.096 +/- 0.016 and 0.064 +/- 0.02 pmol GDP exchanged/min, respectively), but no difference was observed between sedentary and exercised diabetic rats (0.037 +/- 0.001 and 0.044 +/- 0.008 pmol GDP exchanged/min, respectively), and these activities were lower (P < 0.05) than in nondiabetic animals. These data suggest that severe hypoinsulinemia is associated with an inability to increase eIF2B activity in response to exercise. PMID- 11408417 TI - Decreased monocarboxylate transporter 1 in rat soleus and EDL muscles exposed to clenbuterol. AB - We hypothesized that a shift in muscle fiber type induced by clenbuterol would change monocarboxylate transporter 1 (MCT1) content and activity of lactate dehydrogenase (LDH) and isoform pattern and shift myosin heavy chain (MHC) pattern in soleus (Sol) and extensor digitorum longus (EDL) of male rats. In the clenbuterol-administered rats (2.0 mg x kg(-1) x day(-1) subcutaneously for 4 wk), the ratio of muscle weight to body weight increased in the Sol (P < 0.05) and the EDL (P < 0.01). Clenbuterol induced the appearance of fast MHC(2D) and decreased slow MHC(1) in Sol (13%) but had no effect on EDL. The MHC pattern of Sol changed from slow to fast type. Clenbuterol increased LDH-specific activity (P < 0.01) and the ratio of the muscle-type isozyme of LDH to the heart type (P < 0.05) in Sol. The LDH total activity of the EDL muscle was also increased (P < 0.05). Furthermore, MCT1 content significantly (P < 0.05) decreased in both Sol and EDL (27 and 52%, respectively). This study suggests that clenbuterol might mediate the shift of MHC from slow to fast type and the changes in the regulation of lactate metabolism. Novel to this study is the observation that clenbuterol decreases MCT1 content in the hindlimb muscles and that the decrease in MCT1 is not muscle-type specific. It may suggest that the genetic expressions of individual factors involving slow-type MHC, heart-type isozyme of LDH, and MCT1 are associated with one another but are regulated independently. PMID- 11408418 TI - Effect of morning exercise on counterregulatory responses to subsequent, afternoon exercise. AB - The aim of this study was to determine whether a bout of morning exercise (EXE(1)) can alter neuroendocrine and metabolic responses to subsequent afternoon exercise (EXE(2)) and whether these changes follow a gender-specific pattern. Sixteen healthy volunteers (8 men and 8 women, age 27 +/- 1 yr, body mass index 23 +/- 1 kg/m(2), maximal O(2) uptake 31 +/- 2 ml x kg(-1) x min(-1)) were studied after an overnight fast. EXE(1) and EXE(2) each consisted of 90 min of cycling on a stationary bike at 48 +/- 2% of maximal O(2) uptake separated by 3 h. To avoid the confounding effects of hypoglycemia and glycogen depletion, carbohydrate (1.5 g/kg body wt po) was given after EXE(1), and plasma glucose was maintained at euglycemia during both episodes of exercise by a modification of the glucose-clamp technique. Basal insulin levels (7 +/- 1 microU/ml) and exercise-induced insulin decreases (-3 microU/ml) were similar during EXE(1) and EXE(2). Plasma glucose was 5.2 +/- 0.1 and 5.2 +/- 0.1 mmol/l during EXE(1) and EXE(2), respectively. The glucose infusion rate needed to maintain euglycemia during the last 30 min of exercise was increased during EXE(2) compared with EXE(1) (32 +/- 4 vs. 7 +/- 2 micromol x kg(-1) x min(-1)). Although this increased need for exogenous glucose was similar in men and women, gender differences in counterregulatory responses were significant. Compared with EXE(1), epinephrine, norepinephrine, growth hormone, pancreatic polypeptide, and cortisol responses were blunted during EXE(2) in men, but neuroendocrine responses were preserved or increased in women. In summary, morning exercise significantly impaired the body's ability to maintain euglycemia during later exercise of similar intensity and duration. We conclude that antecedent exercise can significantly modify, in a gender-specific fashion, metabolic and neuroendocrine responses to subsequent exercise. PMID- 11408419 TI - Gender alters impact of hypobaric hypoxia on adductor pollicis muscle performance. AB - Recently, we reported that, at similar voluntary force development during static submaximal intermittent contractions of the adductor pollicis muscle, fatigue developed more slowly in women than in men under conditions of normobaric normoxia (NN) (Acta Physiol Scand 167: 233-239, 1999). We postulated that the slower fatigue of women was due, in part, to a greater capacity for muscle oxidative phosphorylation. The present study examined whether a gender difference in adductor pollicis muscle performance also exists during acute exposure to hypobaric hypoxia (HH; 4,300-m altitude). Healthy young men (n = 12) and women (n = 21) performed repeated static contractions at 50% of maximal voluntary contraction (MVC) force of rested muscle for 5 s followed by 5 s of rest until exhaustion. MVC force was measured before and at the end of each minute of exercise and at exhaustion. Exhaustion was defined as an MVC force decline to 50% of that of rested muscle. For each gender, MVC force of rested muscle in HH was not significantly different from that in NN. MVC force tended to decline at a faster rate in HH than in NN for men but not for women. In both environments, MVC force declined faster (P < 0.01) for men than for women. For men, endurance time to exhaustion was shorter (P < 0.01) in HH than in NN [6.08 +/- 0.7 vs. 8.00 +/- 0.7 (SE) min]. However, for women, endurance time to exhaustion was similar (not significant) in HH (12.86 +/- 1.2 min) and NN (13.95 +/- 1.0 min). In both environments, endurance time to exhaustion was longer for women than for men (P < 0.01). Gender differences in the impact of HH on adductor pollicis muscle endurance persisted in a smaller number of men and women matched (n = 4 pairs) for MVC force of rested muscle and thus on submaximal absolute force and, by inference, ATP demand in both environments. In contrast to gender differences in the impact of HH on small-muscle (adductor pollicis) exercise performance, peak O(2) uptake during large-muscle exercise was lower in HH than in NN by a similar (P > 0.05) percentage for men and women (-27.6 +/- 2 and -25.1 +/- 2%, respectively). Our findings are consistent with the postulate of a higher adductor pollicis muscle oxidative capacity in women than in men and imply that isolated performance of muscle with a higher oxidative capacity may be less impaired when the muscle is exposed to HH. PMID- 11408420 TI - Cytokine changes after a marathon race. AB - The influence of carbohydrate (1 l/h of a 6% carbohydrate beverage), gender, and age on pro- and anti-inflammatory plasma cytokine and hormone changes was studied in 98 runners for 1.5 h after two competitive marathon races. The marathoner runners were randomly assigned to carbohydrate (C, n = 48) and placebo (P, n = 50) groups, with beverages administered during the races in a double-blind fashion using color codes. Plasma glucose was higher and cortisol was lower in the C than in the P group after the race (P < 0.001). For all subjects combined, plasma levels of interleukin (IL)-10, IL-1 receptor antagonist (IL-1ra), IL-6, and IL-8 rose significantly immediately after the race and remained above prerace levels 1.5 h later. The pattern of change in all cytokines did not differ significantly between the 12 women and 86 men in the study and the 23 subjects > or =50 yr of age and the 75 subjects <50 yr of age. The pattern of change in IL 10, IL-1ra, and IL-8, but not IL-6, differed significantly between the C and the P group, with higher postrace values measured for IL-10 (109% higher) and IL-1ra (212%) in the P group and for IL-8 (42%) in the C group. In conclusion, plasma levels of IL-10, IL-1ra, IL-6, and IL-8 rose strongly in runners after a competitive marathon, and this was not influenced by age or gender. Carbohydrate ingestion, however, had a major effect in attenuating increases in cortisol and two anti-inflammatory cytokines, IL-10 and IL-1ra. PMID- 11408421 TI - Effects of fat adaptation and carbohydrate restoration on prolonged endurance exercise. AB - We determined the effect of fat adaptation on metabolism and performance during 5 h of cycling in seven competitive athletes who consumed a standard carbohydrate (CHO) diet for 1 day and then either a high-CHO diet (11 g. kg(-1)x day(-1) CHO, 1 g x kg(-1) x day(-1) fat; HCHO) or an isoenergetic high-fat diet (2.6 g x kg( 1) x day(-1) CHO, 4.6 g x kg(-1) x day(-1) fat; fat-adapt) for 6 days. On day 8, subjects consumed a high-CHO diet and rested. On day 9, subjects consumed a preexercise meal and then cycled for 4 h at 65% peak O(2) uptake, followed by a 1 h time trial (TT). Compared with baseline, 6 days of fat-adapt reduced respiratory exchange ratio (RER) with cycling at 65% peak O(2) uptake [0.78 +/- 0.01 (SE) vs. 0.85 +/- 0.02; P < 0.05]. However, RER was restored by 1 day of high-CHO diet, preexercise meal, and CHO ingestion (0.88 +/- 0.01; P < 0.05). RER was higher after HCHO than fat-adapt (0.85 +/- 0.01, 0.89 +/- 0.01, and 0.93 +/- 0.01 for days 2, 8, and 9, respectively; P < 0.05). Fat oxidation during the 4-h ride was greater (171 +/- 32 vs. 119 +/- 38 g; P < 0.05) and CHO oxidation lower (597 +/- 41 vs. 719 +/- 46 g; P < 0.05) after fat-adapt. Power output was 11% higher during the TT after fat-adapt than after HCHO (312 +/- 15 vs. 279 +/- 20 W; P = 0.11). In conclusion, compared with a high-CHO diet, fat oxidation during exercise increased after fat-adapt and remained elevated above baseline even after 1 day of a high-CHO diet and increased CHO availability. However, this study failed to detect a significant benefit of fat adaptation to performance of a 1-h TT undertaken after 4 h of cycling. PMID- 11408422 TI - Blood-brain barrier permeability during dopamine-induced hypertension in fetal sheep. AB - Dopamine is often used as a pressor agent in sick newborn infants, but an increase in arterial blood pressure could disrupt the blood-brain barrier (BBB), especially in the preterm newborn. Using time-dated pregnant sheep, we tested the hypothesis that dopamine-induced hypertension increases fetal BBB permeability and cerebral water content. Barrier permeability was assessed in nine brain regions, including cerebral cortex, caudate, thalamus, brain stem, cerebellum, and spinal cord, by intravenous injection of the small tracer molecule [(14)C]aminoisobutyric acid at 10 min after the start of dopamine or saline infusion. We studied 23 chronically catheterized fetal sheep at 0.6 (93 days, n = 10) and 0.9 (132 days, n = 13) gestation. Intravenous infusion of dopamine increased mean arterial pressure from 38 +/- 3 to 53 +/- 5 mmHg in 93-day fetuses and from 55 +/- 5 to 77 +/- 8 mmHg in 132-day fetuses without a decrease in arterial O(2) content. These 40% increases in arterial pressure are close to the maximum hypertension reported for physiological stresses at these ages in fetal sheep. No significant increases in the brain transfer coefficient of aminoisobutyric acid were detected in any brain region in dopamine-treated fetuses compared with saline controls at 0.6 or 0.9 gestation. There was also no significant increase in cortical water content with dopamine infusion at either age. We conclude that a 40% increase in mean arterial pressure during dopamine infusion in normoxic fetal sheep does not produce substantial BBB disruption or cerebral edema even as early as 0.6 gestation. PMID- 11408423 TI - Role of nitric oxide in the regulation of glucose kinetics in response to endotoxin in dogs. AB - The purpose of the present in vivo study was to determine the role of nitric oxide (NO) in the regulation of glucose metabolism in response to endotoxin by blocking NO synthesis with N(G)-monomethyl-L-arginine (L-NMMA). In five dogs, the appearance and disappearance rates of glucose (by infusion of [6,6 (2)H(2)]glucose), plasma glucose concentration, and plasma hormone concentrations were measured on five different occasions: saline infusion, endotoxin alone (E coli, 1.0 microg/kg i.v.), and endotoxin administration plus three different doses of primed, continuous infusion of L-NMMA. Endotoxin increased rate of appearance of glucose from 13.7 +/- 1.6 to 23.6 +/- 3.3 micromol x kg(-1) x min( 1) (P < 0.05), rate of disappearance of glucose from 13.9 +/- 1.1 to 24.8 +/- 3.1 micromol x kg(-1) x min(-1) (P < 0.001), plasma lactate from 0.5 +/- 0.1 to 1.7 +/- 0.1 mmol/l (P < 0.01), and counterregulatory hormone concentrations. L-NMMA did not affect the rise in rate of appearance and disappearance of glucose, plasma lactate, or the counterregulatory hormone response to endoxin. Plasma glucose levels were not affected by endotoxin with or without L-NMMA. In conclusion, in vivo inhibition of NO synthesis by high doses of L-NMMA does not affect glucose metabolism in response to endotoxin, indicating that NO is not a major mediator of glucose metabolism during endotoxemia in dogs. PMID- 11408424 TI - Effects of postural changes and vestibular lesions on diaphragm and rectus abdominis activity in awake cats. AB - Changes in posture can affect the resting length of the diaphragm, requiring alterations in the activity of both the abdominal muscles and the diaphragm to maintain stable ventilation. To determine the role of the vestibular system in regulating respiratory muscle discharges during postural changes, spontaneous diaphragm and rectus abdominis activity and modulation of the firing of these muscles during nose-up and ear-down tilt were compared before and after removal of labyrinthine inputs in awake cats. In vestibular-intact animals, nose-up and ear-down tilts from the prone position altered rectus abdominis firing, whereas the effects of body rotation on diaphragm activity were not statistically significant. After peripheral vestibular lesions, spontaneous diaphragm and rectus abdominis discharges increased significantly (by approximately 170%), and augmentation of rectus abdominis activity during nose-up body rotation was diminished. However, spontaneous muscle activity and responses to tilt began to recover after a few days after the lesions, presumably because of plasticity in the central vestibular system. These data suggest that the vestibular system provides tonic inhibitory influences on rectus abdominis and the diaphragm and in addition contributes to eliciting increases in abdominal muscle activity during some changes in body orientation. PMID- 11408425 TI - Interactions of exercise training and lipoic acid on skeletal muscle glucose transport in obese Zucker rats. AB - Exercise training (ET) or the antioxidant R(+)-alpha-lipoic acid (R-ALA) individually increases insulin action in the insulin-resistant obese Zucker rat. The purpose of the present study was to determine the interactions of ET and R ALA on insulin action and oxidative stress in skeletal muscle of the obese Zucker rat. Animals either remained sedentary, received R-ALA (30 mg x kg body wt(-1) x day(-1)), performed ET (treadmill running), or underwent both R-ALA treatment and ET for 6 wk. During an oral glucose tolerance test, ET alone or in combination with R-ALA resulted in a significant lowering of the glucose (26-32%) and insulin (29-30%) responses compared with sedentary controls. R-ALA alone decreased (19%) the glucose-insulin index (indicative of increased insulin sensitivity), and this parameter was reduced (48-52%) to the greatest extent in the ET and combined treatment groups. ET or R-ALA individually increased insulin-mediated glucose transport activity in isolated epitrochlearis (44-48%) and soleus (37-57%) muscles. The greatest increases in insulin action in these muscles (80 and 99%, respectively) were observed in the combined treatment group. Whereas the improvement in insulin-mediated glucose transport in soleus due to R-ALA was associated with decreased protein carbonyl levels (an index of oxidative stress), improvement because of ET was associated with decreased protein carbonyls as well as enhanced GLUT-4 protein. However, there was no interactive effect of ET and R ALA on GLUT-4 protein or protein carbonyl levels. These results indicate that ET and R-ALA interact in an additive fashion to improve insulin action in insulin resistant skeletal muscle. Because the further improvement in muscle glucose transport in the combined group was not associated with additional upregulation of GLUT-4 protein or a further reduction in oxidative stress, the mechanism for this interaction must be due to additional, as yet unidentified, factors. PMID- 11408426 TI - Chamber properties from transmitral flow: prediction of average and passive left ventricular diastolic stiffness. AB - A chamber stiffness (K(LV))-transmitral flow (E-wave) deceleration time relation has been invasively validated in dogs with the use of average stiffness [(DeltaP/DeltaV)(avg)]. K(LV) is equivalent to k(E), the (E-wave) stiffness of the parameterized diastolic filling model. Prediction and validation of 1) (DeltaP/DeltaV)(avg) in terms of k(E), 2) early rapid-filling stiffness [(DeltaP/DeltaV)(E)] in terms of k(E), and 3) passive (postdiastasis) chamber stiffness [(DeltaP/DeltaV)(PD)] from A waves in terms of the stiffness parameter for the Doppler A wave (k(A)) have not been achieved. Simultaneous micromanometric left ventricular (LV) pressure (LVP) and transmitral flow from 131 subjects were analyzed. (DeltaP)(avg) and (DeltaV)(avg) utilized the minimum LVP-LV end-diastolic pressure interval. (DeltaP/DeltaV)(E) utilized DeltaP and DeltaV from minimum LVP to E-wave termination. (DeltaP/DeltaV)(PD) utilized atrial systolic DeltaP and DeltaV. E- and A-wave analysis generated k(E) and k(A). For all subjects, noninvasive-invasive relations yielded the following equations: k(E) = 1,401. (DeltaP/DeltaV)(avg) + 59.2 (r = 0.84) and k(E) = 229.0. (DeltaP/DeltaV)(E) + 112 (r = 0.80). For subjects with diastasis (n = 113), k(A) = 1,640. (DeltaP/DeltaV)(PD) - 8.40 (r = 0.89). As predicted, k(A) showed excellent correlation with (DeltaP/DeltaV)(PD); k(E) correlated highly with (DeltaP/DeltaV)(avg). In vivo validation of average, early, and passive chamber stiffness facilitates quantitative, noninvasive diastolic function assessment from transmitral flow. PMID- 11408427 TI - Growth hormone pulsatility profile characteristics following acute heavy resistance exercise. AB - This investigation examined the hypothesis that acute heavy resistance exercise (AHRE) would increase overnight concentrations of circulating human growth hormone (hGH). Ten men (22 +/- 1 yr, 177 +/- 2 cm, 79 +/- 3 kg, 11 +/- 1% body fat) underwent two overnight blood draws sampled every 10 min from 1700 to 0600: a control and an AHRE condition. The AHRE was conducted from 1500 to 1700 and was a high-volume, multiset exercise bout. Three different immunoassays measured hGH concentrations: the Nichols immunoradiometric assay (Nichols IRMA), National Institute of Diabetes and Digestive and Kidney Diseases radioimmunoassay (NIDDK RIA), and the Diagnostic Systems Laboratory immunofunctional assay (DSL IFA). The Pulsar peak detection system was used to evaluate the pulsatility profile characteristics of hGH. Maximum hGH was lower in the exercise (10.7 microg/l) vs. the control (15.4 microg/l) condition. Mean pulse amplitude was lower in the exercise vs. control condition when measured by the Nichols IRMA and the DSL IFA. A differential pattern of release was also observed after exercise in which hGH was lower in the first half of sleep but higher in the second half. We conclude that AHRE does influence the temporal pattern of overnight hGH pulsatility. Additionally, because of the unique molecular basis of the DSL IFA, this influence does have biological relevance because functionally intact molecules are affected. PMID- 11408428 TI - Molecular adaptations in human skeletal muscle to endurance training under simulated hypoxic conditions. AB - This study was performed to explore changes in gene expression as a consequence of exercise training at two levels of intensity under normoxic and normobaric hypoxic conditions (corresponding to an altitude of 3,850 m). Four groups of human subjects trained five times a week for a total of 6 wk on a bicycle ergometer. Muscle biopsies were taken, and performance tests were carried out before and after the training period. Similar increases in maximal O(2) uptake (8.3-13.1%) and maximal power output (11.4-20.8%) were found in all groups. RT PCR revealed elevated mRNA concentrations of the alpha-subunit of hypoxia inducible factor 1 (HIF-1) after both high- (+82.4%) and low (+78.4%)-intensity training under hypoxic conditions. The mRNA of HIF-1alpha(736), a splice variant of HIF-1alpha newly detected in human skeletal muscle, was shown to be changed in a similar pattern as HIF-1alpha. Increased mRNA contents of myoglobin (+72.2%) and vascular endothelial growth factor (+52.4%) were evoked only after high intensity training in hypoxia. Augmented mRNA levels of oxidative enzymes, phosphofructokinase, and heat shock protein 70 were found after high-intensity training under both hypoxic and normoxic conditions. Our findings suggest that HIF-1 is specifically involved in the regulation of muscle adaptations after hypoxia training. Fine-tuning of the training response is recognized at the molecular level, and with less sensitivity also at the structural level, but not at global functional responses like maximal O(2) uptake or maximal power output. PMID- 11408429 TI - Muscle regeneration during hindlimb unloading results in a reduction in muscle size after reloading. AB - The hindlimb-unloading model was used to study the ability of muscle injured in a weightless environment to recover after reloading. Satellite cell mitotic activity and DNA unit size were determined in injured and intact soleus muscles from hindlimb-unloaded and age-matched weight-bearing rats at the conclusion of 28 days of hindlimb unloading, 2 wk after reloading, and 9 wk after reloading. The body weights of hindlimb-unloaded rats were significantly (P < 0.05) less than those of weight-bearing rats at the conclusion of hindlimb unloading, but they were the same (P > 0.05) as those of weight-bearing rats 2 and 9 wk after reloading. The soleus muscle weight, soleus muscle weight-to-body weight ratio, myofiber diameter, number of nuclei per millimeter, and DNA unit size were significantly (P < 0.05) smaller for the injured soleus muscles from hindlimb unloaded rats than for the soleus muscles from weight-bearing rats at each recovery time. Satellite cell mitotic activity was significantly (P < 0.05) higher in the injured soleus muscles from hindlimb-unloaded rats than from weight bearing rats 2 wk after reloading, but it was the same (P > 0.05) as in the injured soleus muscles from weight-bearing rats 9 wk after reloading. The injured soleus muscles from hindlimb-unloaded rats failed to achieve weight-bearing muscle size 9 wk after reloading, because incomplete compensation for the decrease in myonuclear accretion and DNA unit size expansion occurred during the unloading period. PMID- 11408430 TI - pH heterogeneity in tibial anterior muscle during isometric activity studied by (31)P-NMR spectroscopy. AB - The occurrence of pH heterogeneity in human tibial anterior muscle during sustained isometric exercise is demonstrated by applying (31)P-nuclear magnetic resonance (NMR) spectroscopy in a study of seven healthy subjects. Exercise was performed at 30 and 60% of maximal voluntary contraction (MVC) until fatigue. The NMR spectra, as localized by a surface coil and improved by proton irradiation, were obtained at a high time resolution (16 s). They revealed the simultaneous presence of two pH pools during most experiments. Maximum difference in the two pH levels during exercise was 0.40 +/- 0.07 (30% MVC, n = 7) and 0.41 +/- 0.03 (60% MVC, n = 3). Complementary two-dimensional (31)P spectroscopic imaging experiments in one subject supported the supposition that the distinct pH pools reflect the metabolic status of the main muscle fiber types. The relative size of the P(i) peak in the spectrum attributed to the type II fiber pool increases with decreasing pH levels. This phenomenon is discussed in the context of the size principle stating that the smaller (type I) motor units are recruited first. PMID- 11408431 TI - General characteristics of the sigmoidal model equation representing quasi-static pulmonary P-V curves. AB - A pulmonary pressure-volume (P-V) curve represented by a sigmoidal model equation with four parameters, V(P) = a + b[1 + exp[-(P - c)/d]](-1), has been demonstrated to fit inflation and deflation data obtained under a variety of conditions extremely well. In the present report, a differential equation on V(P) is identified, thus relating the fourth parameter, d, to the difference between the upper and the lower asymptotes of the volume, b, through a proportionality constant, alpha, with its order of magnitude of 10(-4) to 10(-5) (in ml(-1). cmH(2)O(-1)). When the model equation is normalized using a nondimensional volume, (-1 < < 1), and a nondimensional pressure, (=(p/c) - 1), the resulting - curve depends on a single nondimensional parameter, Lambda = alphabc. A nondimensional work of expansion/compression, (1-2), is also obtained along the quasi-static sigmoidal P-V curve between an initial volume (at 1) and a final volume (at 2). Six sets of P-V data available in the literature are used to show the changes that occur in these two parameters (Lambda defining the shape of the sigmoidal curve and (1-2) accounting for the range of clinical data) with different conditions of the total respiratory system. The clinical usefulness of these parameters requires further study. PMID- 11408432 TI - Further analysis of transcytosis of free polypeptides and polypeptide-coated nanobeads in rabbit nasal mucosa. AB - In rabbit nasal mucosa, free polypeptides and polypeptide-coated nanospheres are actively absorbed by the M cells present in specialized areas of the epithelium. Because polypeptide-coated nanosphere transport was abolished in the presence of free polypeptides, free polypeptides and polypeptide-coated nanospheres are shown here to compete. Fluxes of polypeptide-coated nanospheres with 356, 490, and 548 nm diameters have been compared. BSA-coated beads were poorly transported, at the same rate, when bead diameters were 356 or 490 nm [net flux of approximately 2 2.5 x 10(6) nanospheres (nan). cm(-2) x h(-1)]; however, their net transport largely increased toward a value of 25 x 10(6) nan. cm(-2) x h(-1) at a diameter of 548 nm. Insulin-coated beads displayed a net flux that was significantly higher than BSA-coated beads but equally were transported at the same rate (net flux of approximately 8.0 x 10(6) nan. cm(-2) x h(-1)) at diameters of 356 or 490 nm; once again, their net flux significantly increased toward a value of 25 x 10(6) nan. cm(-2) x h(-1), if the bead diameter was 548 nm. Insulin plus anti insulin IgG-coated 490-nm-diameter beads displayed a very high net flux, although not yet saturating (approximately 60 x 10(6) nan. cm(-2) x h(-1)); however, a significantly lower saturated net flux (once again approximately 25 x 10(6) nan. cm(-2) x h(-1)) was shown with 548-nm-diameter beads. In conclusion, 1) in the range of 356-490 nm diameter, net transport was independent of bead diameter and, conversely, largely dependent on the coating polypeptides, and 2) at 548 nm diameter, nanospheres tended to be transferred at similar rates independently of coating kind and the maximal net transport capacity of the mucosa was reduced. The suspension viscosity largely increased with 548-nm polypeptide-coated nanospheres; this fact is hypothetically proposed to be the cause of these events. PMID- 11408433 TI - Validity of inspiratory and expiratory methods of measuring gas exchange with a computerized system. AB - The accuracy of a computerized metabolic system, using inspiratory and expiratory methods of measuring ventilation, was assessed in eight male subjects. Gas exchange was measured at rest and during five stages on a cycle ergometer. Pneumotachometers were placed on the inspired and expired side to measure inspired (VI) and expired ventilation (VE). The devices were connected to two systems sampling expired O(2) and CO(2) from a single mixing chamber. Simultaneously, the criterion (Douglas bag, or DB) method assessed VE and fractions of O(2) and CO(2) in expired gas (FE(O(2)) and FE(CO(2))) for subsequent calculation of O(2) uptake (VO(2)), CO(2) production (VCO(2)), and respiratory exchange ratio. Both systems accurately measured metabolic variables over a wide range of intensities. Though differences were found between the DB and computerized systems for FE(O(2)) (both inspired and expired systems), FE(CO(2)) (expired system only), and VO(2) (inspired system only), the differences were extremely small (FE(O(2)) = 0.0004, FE(CO(2)) = -0.0003, VO(2) = -0.018 l/min). Thus a computerized system, using inspiratory or expiratory configurations, permits extremely precise measurements to be made in a less time consuming manner than the DB technique. PMID- 11408434 TI - Gender differences in carbohydrate loading are related to energy intake. AB - We demonstrated that female endurance athletes did not increase their muscle glycogen concentration after an increase in the dietary carbohydrate intake (58 - > 74%), whereas men did (Tarnopolsky MA, SA Atkinson, SM Phillips, and JD McDougall, J Appl Physiol 78: 1360-1368, 1995). This may have been related to a lower energy or carbohydrate intake by the women or due to an inherent gender difference in glycogen storage capacity. We examined whether well-trained men (n = 6) and women (n = 6) increased muscle glycogen concentration after an increase in both the relative (58 --> 75%) and absolute energy and carbohydrate intake and whether potential gender differences were related to muscle hexokinase enzyme activity. Subjects were randomly allocated to three diets [Hab, habitual; CHO, high carbohydrate (75%); and CHO + E, extra energy + CHO ( upward arrow~34%)] for a 4-day period before a muscle biopsy for analysis of total and pro- and macroglycogen and hexokinase activity. Total glycogen concentration was higher for the men on the CHO and CHO + E trials compared with Hab (P < 0.05), whereas women increased only on the CHO + E trial compared with Hab (P < 0.05). There were no gender differences in the proportion of pro- and macroglycogen or hexokinase activity. A low energy intake may explain the previously reported lower capacity for women to glycogen load compared with men. PMID- 11408435 TI - Purine salvage to adenine nucleotides in different skeletal muscle fiber types. AB - Rates of purine salvage of adenine and hypoxanthine into the adenine nucleotide (AdN) pool of the different skeletal muscle phenotype sections of the rat were measured using an isolated perfused hindlimb preparation. Tissue adenine and hypoxanthine concentrations and specific activities were controlled over a broad range of purine concentrations, ranging from 3 to 100 times normal, by employing an isolated rat hindlimb preparation perfused at a high flow rate. Incorporation of [(3)H]adenine or [(3)H]hypoxanthine into the AdN pool was not meaningfully influenced by tissue purine concentration over the range evaluated (approximately 0.10-1.6 micromol/g). Purine salvage rates were greater (P < 0.05) for adenine than for hypoxanthine (35-55 and 20-30 nmol x h(-1) x g(-1), respectively) and moderately different (P < 0.05) among fiber types. The low-oxidative fast-twitch white muscle section exhibited relatively low rates of purine salvage that were approximately 65% of rates in the high-oxidative fast-twitch red section of the gastrocnemius. The soleus muscle, characterized by slow-twitch red fibers, exhibited a high rate of adenine salvage but a low rate of hypoxanthine salvage. Addition of ribose to the perfusion medium increased salvage of adenine (up to 3- to 6-fold, P < 0.001) and hypoxanthine (up to 6- to 8-fold, P < 0.001), depending on fiber type, over a range of concentrations up to 10 mM. This is consistent with tissue 5-phosphoribosyl-1-pyrophosphate being rate limiting for purine salvage. Purine salvage is favored over de novo synthesis, inasmuch as delivery of adenine to the muscle decreased (P < 0.005) de novo synthesis of AdN. Providing ribose did not alter this preference of purine salvage pathway over de novo synthesis of AdN. In the absence of ribose supplementation, purine salvage rates are relatively low, especially compared with the AdN pool size in skeletal muscle. PMID- 11408436 TI - Effect of REM sleep on retroglossal cross-sectional area and compliance in normal subjects. AB - It has been proposed that the upper airway compliance should be highest during rapid eye movement (REM) sleep. Evidence suggests that the increased compliance is secondary to an increased retroglossal compliance. To test this hypothesis, we examined the effect of sleep stage on the relationship of retroglossal cross sectional area (CSA; visualized with a fiber-optic scope) to pharyngeal pressure measured at the level of the oropharynx during eupneic breathing in subjects without significant sleep-disordered breathing. Breaths during REM sleep were divided into phasic (associated with eye movement, PREM) and tonic (not associated with eye movements, TREM). Retroglossal CSA decreased with non-REM (NREM) sleep and decreased further in PREM [wake 156.8 +/- 48.6 mm(2), NREM 104.6 +/- 65.0 mm(2) (P < 0.05 wake vs. NREM), TREM 83.1 +/- 46.4 mm(2) (P = not significant NREM vs. TREM), PREM 73.9 + 39.2 mm(2) (P < 0.05 TREM vs. PREM)]. Retroglossal compliance, defined as the slope of the regression CSA vs. pharyngeal pressure, was the same between all four conditions (wake -0.7 + 2.1 mm(2)/cmH(2)O, NREM 0.6 +/- 3.0 mm(2)/cmH(2)O, TREM -0.2 +/- 3.3 mm(2)/cmH(2)O, PREM -0.6 +/- 5.1 mm(2)/cmH(2)O, P = not significant). We conclude that the intrinsic properties of the airway wall determine retroglossal compliance independent of changes in the neuromuscular activity associated with changes in sleep state. PMID- 11408437 TI - Effects of 3-day bed rest on physiological responses to graded exercise in athletes and sedentary men. AB - To test the hypotheses that short-term bed-rest (BR) deconditioning influences metabolic, cardiorespiratory, and neurohormonal responses to exercise and that these effects depend on the subjects' training status, 12 sedentary men and 10 endurance- and 10 strength-trained athletes were submitted to 3-day BR. Before and after BR they performed incremental exercise test until volitional exhaustion. Respiratory gas exchange and heart rate (HR) were recorded continuously, and stroke volume (SV) was measured at submaximal loads. Blood was taken for lactate concentration ([LA]), epinephrine concentration ([Epi]), norepinephrine concentration ([NE]), plasma renin activity (PRA), human growth hormone concentration ([hGH]), testosterone, and cortisol determination. Reduction of peak oxygen uptake (VO(2 peak)) after BR was greater in the endurance athletes than in the remaining groups (17 vs. 10%). Decrements in VO(2 peak) correlated positively with the initial values (r = 0.73, P < 0.001). Resting and exercise respiratory exchange ratios were increased in athletes. Cardiac output was unchanged by BR in all groups, but exercise HR was increased and SV diminished in the sedentary subjects. The submaximal [LA] and [LA] thresholds were decreased in the endurance athletes from 71 to 60% VO(2 peak) (P < 0.001); they also had an earlier increase in [NE], an attenuated increase in [hGH], and accentuated PRA and cortisol elevations during exercise. These effects were insignificant in the remaining subjects. In conclusion, reduction of exercise performance and modifications in neurohormonal response to exercise after BR depend on the previous level and mode of physical training, being the most pronounced in the endurance athletes. PMID- 11408438 TI - Regulation of skeletal muscle ATP catabolism by AMPD1 genotype during sprint exercise in asymptomatic subjects. AB - Deficiency of myoadenylate deaminase, the muscle isoform of AMP deaminase encoded by the AMPD1 gene, is a common myopathic condition associated with alterations in skeletal muscle energy metabolism. However, recent studies have demonstrated that most individuals harboring this genetic abnormality are asymptomatic. Therefore, 18 healthy subjects with different AMPD1 genotypes were studied during a 30-s Wingate test in order to evaluate the influence of this inherited defect in AMPD1 expression on skeletal muscle energy metabolism and exercise performance in the asymptomatic population. Exercise performances were similar across the AMPD1 genotypes, whereas significant differences in several descriptors of energy metabolism were observed. Normal homozygotes (NN) exhibited the highest levels of AMP deaminase activities, net ATP catabolism, and IMP accumulation, whereas intermediate values were observed in heterozygotes (MN). Conversely, mutant homozygotes (MM) had very low AMP deaminase activities and showed no significant net catabolism of ATP or IMP accumulation. Accordingly, MM also did not show any postexercise increase in plasma ammonia. Unexpectedly, MN consistently exhibited greater increases in plasma ammonia compared with NN despite the relatively lower accumulation of IMP in skeletal muscle. Moreover, time course profiles of postexercise plasma ammonia and blood lactate accumulation also differed across AMPD1 genotypes. Finally, analysis of adenosine in leftover biopsy material revealed a modest twofold increase in MN and a dramatic 25-fold increase in MM. PMID- 11408439 TI - Pronounced effect of minor changes in body temperature on ischemia and reperfusion injury in rat liver. AB - This study examined the effects of 1 degrees C hypo- or hyperthermia on in vivo liver ischemia and reperfusion (I/R) injury in 15 fasted male Wistar rats. Rats were ventilated, and rectal temperature was maintained at 36, 37 (normothermic), or 38 degrees C. In all rats, 70% liver ischemia was induced by clamping the afferent vessels to the median and left lateral lobes for 60 min, and reperfusion was allowed for 90 min. Changes in plasma aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alpha-glutathione S-transferase (alpha-GST) levels were measured, hemodynamics and bile secretion were monitored, and arterial blood-gas analysis was performed. All ventilated rats showed a normal pH, arterial PCO(2), and arterial PO(2). AST, ALT, and alpha-GST levels were significantly higher in the 38 degrees C group when compared with the 36 and 37 degrees C groups after ischemia. No differences in bile secretion were found between all groups. Histopathological alterations were in agreement with AST, ALT, and alpha-GST levels in plasma. We conclude that a decrease of only 1 degrees C in body temperature significantly attenuates liver I/R injury, whereas an increase of 1 degrees C significantly increases liver I/R injury. PMID- 11408440 TI - Role of speed vs. grade in relation to muscle pump function at locomotion onset. AB - We sought to clarify the roles of contraction frequency (speed) and contraction force (grade) in the rise in muscle blood flow at the onset of locomotion. Shoemaker et al. (Can J Physiol Pharmacol 76: 418-427, 1998) explored this relationship in human handgrip exercise and found that the time course of the rise in muscle vascular conductance was similar when a light weight was lifted in a fast cadence and a heavy weight was lifted in a slow cadence (total work constant). This indicates that muscle pumping (contraction frequency) was of limited importance in governing the time course. Rather, vasodilator substances released in proportion to the total work performed appeared to determine the pattern and extent of the rise in conductance. We hypothesized that conductance would rise faster during locomotion at a high speed (frequency) and low grade (force) than at a low speed and high grade, despite similar total increases in conductance, owing to more effective muscle pumping at faster contraction rates. Seven male rats performed nine 1-min bouts of treadmill locomotion across a combination of three speeds (5, 10, and 20 m/min) and three grades (-10, 0, and +15 degrees ) in random order. Locomotion at 10 m/min and 0 degrees grade and 20 m/min and -10 degrees grade led to an equal rise in terminal aortic vascular conductance. However, the equal rise was achieved more quickly at the higher running speed, suggestive of more effective muscle pumping. Across the nine combinations of exercise, speed began to exert a statistically significant influence on conductance by the 3rd s of locomotion. Grade did not begin to exert an influence until the 12th s of locomotion (similar to the delays reported for arteriolar dilation to muscle contraction). Additional experiments in dogs provided similar results. Thus the muscle pump appears to initiate the increase in blood flow in proportion to contraction frequency at locomotion onset. PMID- 11408441 TI - Influences of repetitive muscle contractions with different modes on tendon elasticity in vivo. AB - The present study aimed to investigate the effects of repetitive muscle contractions on the elasticity of human tendon structures in vivo. Before and after each endurance test, the elongation of the tendon and aponeurosis of vastus lateralis muscle (L) was directly measured by ultrasonography while the subjects performed ramp isometric knee extension up to maximal voluntary isometric contraction (MVC). Six male subjects performed muscle endurance tests that consisted of knee extension tasks with four different contraction modes: 1) 50 repetitions of maximal voluntary eccentric action for 3 s with 3 s of relaxation (ET1), 2) three sets of 50 repetitions of MVC for 1 s with 3 s of relaxation (ET2), 3) 50 repetitions of MVC for 3 s with 3 s of relaxation (ET3), and 4) 50 repetitions of 50% MVC for 6 s with 6 s of relaxation (ET4). In ET1 and ET2, there were no significant differences in L values at any force production levels between before and after endurance tests. In the cases of ET3 and ET4, however, the extent of elongation after the completion of the tests tended to be greater. The L values above 330 N in ET3 and 440 N in ET4, respectively, were significantly greater after endurance tests than before. These results suggested that the repeated longer duration contractions would make the tendon structures more compliant and that the changes in the elasticity might be not be affected by either muscle action mode or force production level but by the duration of action. PMID- 11408442 TI - Time delay of vagally mediated cardiac baroreflex response varies with autonomic cardiovascular control. AB - To examine whether changes in autonomic activity have an effect on the latency of the vagally mediated cardiac baroreflex response in humans, we investigated the effects of neck suction fluctuating sinusoidally at 0.2 Hz on R-R intervals (known to be mediated mainly by vagal activity) in the supine position, during 15 degrees head-down tilt and 60 degrees head-up tilt, and during vagotonic (2 microg/kg) and vagolytic (10 microg/kg) doses of atropine while the subjects breathed at 0.25 Hz. The phase shift between fluctuations in neck chamber pressure and in R-R interval was calculated by complex transfer function analysis and was used as a measure of the time delay between carotid baroreceptor stimulation and cardiac effector response. Cardiac baroreflex responsiveness increased significantly during low-dose atropine and decreased during head-up tilt or 10 microg/kg atropine. With increasing tilt angle, the time delay between cyclic baroreceptor stimulation and oscillations in R-R interval increased from 0.32 +/- 0.27 s (head down), to 0.59 +/- 0.25 s (supine position, P < 0.05 vs. head down), and to 0.86 +/- 0.27 s (head up, P < 0.01 vs. supine). Low-dose atropine had a similar effect to head-down tilt on baroreflex latency, whereas 10 microg/kg atropine increased the time delay markedly to 1.24 +/- 0.30 s. Our results demonstrate that changes in autonomic activity, generated either by gravitational stimulus or by atropine, not only affect baroreflex responsiveness but also have a major influence on the latency of the vagally mediated carotid baroreceptor-heart rate reflex. The prolonged baroreflex latency during decreased parasympathetic function may contribute to an unstable regulation of heart rate in patients with cardiac disease. PMID- 11408443 TI - Analysis of tracheal mechanics and applications. AB - We have developed a mathematical model for a tracheal ring that consists of a "horseshoe" of cartilage with its tips joined by a membrane. The ring is subjected to a uniform transmural pressure (Ptm) difference. The model was used to calculate the cross-sectional area (A) of the trachea. Whereas the mechanics of the deformation of the cartilage were analyzed using elastica theory, the posterior membrane was treated as a simple membrane that is inextensible under changes in Ptm. The membrane can be specified to be of any length less than baseline and thus can represent a posterior membrane under tension. The cartilage can have specifiable nonuniform unstressed curvature as well as nonuniform bending stiffness. We have investigated the effect on the tracheal A-Ptm curve of posterior membrane length and tensile force in the membrane, cartilage shape and elasticity, and localized weakening of the cartilage. The model predictions are in good agreement with magnetic resonance imaging data from rabbit tracheas and show that the shape of the horseshoe as well as the posterior membrane force are important determinants of tracheal compliance. PMID- 11408444 TI - Dynamic effects of positive-pressure ventilation on canine left ventricular pressure-volume relations. AB - Positive-pressure ventilation (PPV) may affect left ventricular (LV) performance by altering both LV diastolic compliance and pericardial pressure (Ppc). We measured the effect of PPV on LV intraluminal pressure, Ppc, LV volume, and LV cross-sectional area in 17 acute anesthetized dogs. To account for changes in lung volume independent of changes in Ppc and differences in contractility, measures were made during both open- and closed-chest conditions, during closed chest with and without chest wall binding, and after propranolol-induced acute ventricular failure (AVF). Apneic end-systolic pressure-volume relations (ESPVR) were generated by inferior vena caval occlusions. With the open chest, PPV had no effects. With the chest closed, PPV inspiration decreased LV end-diastolic volume (EDV) along its diastolic compliance curve and decreased end-systolic volume (ESV) such that the end-systolic pressure-volume domain was shifted to a point left of the LV ESPVR, even when referenced to Ppc. The decrease in EDV was greater in control than in AVF conditions, whereas the shift of the ESV to the left of the ESPVR was greater with AVF than in control conditions. We conclude that the hemodynamic effects of PPV inspiration are due primarily to changes in intrathoracic pressure and that the inspiration-induced decreases of LV EDV reflect direct effects of intrathoracic pressure on LV filling. The decreases in LV ESV exceed the amount explained solely by a reduction in LV ejection pressure. PMID- 11408445 TI - Exercise training increases the Ca(2+) sensitivity of tension in rat cardiac myocytes. AB - The heart is known to respond to a program of chronic exercise in ways that enhance cardiac function. However, the cellular mechanisms involved in training induced improvements in the contractile function of the myocardium are not known. In this study we tested the hypothesis that increased contractility of the myocardium associated with exercise training is due, in part, to increases in the Ca(2+) sensitivity of steady-state tension. Female Sprague-Dawley rats were randomly divided into sedentary control (C) and exercise-trained (T) groups. The T rats underwent 11 wk of progressive treadmill exercise (1 h/day, 5 days/wk, 26 m/min, 20% grade). Evidence of training effect included a 5.9% increase in heart mass, increases in heart weight-to-body weight ratio, and a 60% increase in skeletal muscle citrate synthase activity in T rats compared with C rats. After the training program, cardiac myocytes were isolated from T and C hearts. Myocytes were chemically skinned (i.e., the sarcolemma was removed) and attached to a force transducer, and steady-state tension was determined in solutions of various Ca(2+) concentrations ([Ca(2+)]). Myocytes isolated from the hearts of T rats showed a significantly (P < 0.01) increased sensitivity of tension to [Ca(2+)]. The [Ca(2+)] giving 50% of maximal tension (pCa(50)) was 5.90 +/- 0.033 and 5.82 +/- 0.023 (SD) in T and C myocytes, respectively (n = 70 myocytes/group). This result suggests that exercise training affects the myofibrillar proteins, such that Ca(2+) sensitivity is increased, and that this may be the mechanism that underlies, at least in part, the effect of training to increase myocardial contractility. PMID- 11408446 TI - Ultrasound measurements of fetal breathing movements in the rat. AB - The goal of this study was to determine when fetal breathing movements (FBMs) commence in the rat and to characterize age-dependent changes of FBMs in utero. These data provide a frame of reference for parallel in vitro studies of the cellular, synaptic, and network properties of the perinatal rat respiratory system. Ultrasound recordings were made from unanesthetized Sprague-Dawley rats from embryonic (E) day 15 (E15) to E20. Furthermore, the effects of respiratory stimulants (doxapram and aminophylline) and hypoxia on FBMs were studied. Single FBMs, occurring at a very low frequency (approximately 8 FBMs/h), commenced at E16. The incidence of single FBMs increased to approximately 80 FBMs/h by E20. Episodes of clustered rhythmic FBMs were first observed at E18 (approximately 40 FBMs/h). The incidence of episodic clustered FBMs increased to approximately 300 FMBs/h by E20, with the duration of each episode ranging from approximately 40 to 180 s. Doxapram, presumably acting to stimulate carotid body receptors, did not increase FBMs until E20, when the incidence of episodic clustered FBMs increased twofold. Aminophylline, a central-acting stimulant, caused an increase in episodic clustered FBMs after E17, reaching significance at E20 (3-fold increase). Exposing the dam to 10% O(2) caused a rapid, marked suppression of FBMs (5-fold decrease) that was readily reversed on exposure to room air. PMID- 11408447 TI - Microvascular and interstitial PO(2) measurements in rat skeletal muscle by phosphorescence quenching. AB - To clarify the transport of O(2) across the microvessels in skeletal muscle, we designed an intravital laser microscope that utilizes a phosphorescence quenching technique to determine both the microvascular and tissue PO(2). After we injected the phosphorescent probe into systemic blood, phosphorescence excited by a N(2) dye pulse laser was detected with a photomultiplier over a 10 microm in diameter area. In vitro and in vivo calibrations confirmed that the present method is accurate for PO(2) measurements in the range of 7-90 Torr (r = 0.958) and has a rapid response time. This method was then used to measure the PO(2) of microvessels with different diameters (40-130 microm) and of interstitial spaces in rat cremaster muscle. These measurements showed a significant drop in PO(2) in the arterioles after branching (from 74.6 to 46.6 Torr) and the presence of a large PO(2) gradient at the blood-tissue interface of arterioles (15-20 Torr). These findings suggest that capillaries are not the sole source of oxygen supply to surrounding tissue. PMID- 11408448 TI - Carotid body denervation in dogs: eupnea and the ventilatory response to hyperoxic hypercapnia. AB - We assessed the time course of changes in eupneic arterial PCO(2) (Pa(CO(2))) and the ventilatory response to hyperoxic rebreathing after removal of the carotid bodies (CBX) in awake female dogs. Elimination of the ventilatory response to bolus intravenous injections of NaCN was used to confirm CBX status on each day of data collection. Relative to eupneic control (Pa(CO(2)) = 40 +/- 3 Torr), all seven dogs hypoventilated after CBX, reaching a maximum Pa(CO(2)) of 53 +/- 6 Torr by day 3 post-CBX. There was no significant recovery of eupneic Pa(CO(2)) over the ensuing 18 days. Relative to control, the hyperoxic CO(2) ventilatory (change in inspired minute ventilation/change in end-tidal PCO(2)) and tidal volume (change in tidal volume/ change in end-tidal PCO(2)) response slopes were decreased 40 +/- 15 and 35 +/- 20% by day 2 post-CBX. There was no recovery in the ventilatory or tidal volume response slopes to hyperoxic hypercapnia over the ensuing 19 days. We conclude that 1) the carotid bodies contribute approximately 40% of the eupneic drive to breathe and the ventilatory response to hyperoxic hypercapnia and 2) there is no recovery in the eupneic drive to breathe or the ventilatory response to hyperoxic hypercapnia after removal of the carotid chemoreceptors, indicating a lack of central or aortic chemoreceptor plasticity in the adult dog after CBX. PMID- 11408449 TI - Periarticular cancellous bone changes following anterior cruciate ligament injury. AB - To understand more fully the early bone changes in an experimental model of osteoarthrosis, we quantified periarticular bone mineral density and bone mechanical properties in anterior cruciate ligament transected (ACLX) knee joints (4, 10, 32, and 39 wk post-ACLX) compared with contralateral joints and unoperated normal joints of skeletally mature animals. Maximal stress and energy were significantly reduced in ACLX cancellous bone from the medial femoral condyles at 4 wk postinjury. All mechanical properties (e.g., yield stress and elastic modulus) declined after 4 wk and were significantly reduced at 10 wk. ACLX bone mineral density was significantly reduced at all measured time points. Ash content was significantly reduced at 10 and 32 wk. Changes in the lateral condyles were similar but less pronounced than in the medial condyles. These bony changes accompanied the earliest articular cartilage molecular changes and preceded changes in the articular cartilage gross morphology. We suggest that these early changes in bone mechanical behavior contribute to the progression of osteoarthrosis and pathogenic changes in the joint. PMID- 11408450 TI - Adenosine A(1) and A(2A) receptors modulate sleep state and breathing in fetal sheep. AB - This study was designed to determine the adenosine (Ado) receptor subtype that mediates the depressant effects of Ado on fetal breathing and rapid eye movements (REM). In chronically catheterized fetal sheep (>0.8 term), intra-arterial infusion of N(6)-cyclopentyladenosine (CPA), an Ado A(1)-receptor agonist, increased the incidence of high-voltage electrocortical (ECoG) activity while virtually abolishing low-voltage activity, REM, and breathing. These effects were blocked by 9-cyclopentyl-1,3-dipropylxanthine (DPCPX), an Ado A(1)-receptor antagonist. Infusion of DPCPX alone increased breath amplitude but had no significant effect on inspiratory duration, breath interval, incidence of REM, or incidence of low-voltage activity. Ado A(2A)-receptor blockade with ZM-241385 increased the incidence of low-voltage ECoG activity, REM, and breathing but had no effect on breath amplitude or respiratory cycle. Both DPCPX and ZM-241385 eliminated the inhibitory effects of Ado on REM and breathing. We conclude that 1) Ado A(1) receptors tonically inhibit fetal respiratory drive, 2) Ado A(2A) receptors tonically inhibit REM-like behavioral state, and 3) both Ado A(1) and A(2A) receptors mediate the depressant effects of Ado on REM and breathing. PMID- 11408451 TI - Effects of Pseudomonas aeruginosa endotoxin on vasodilation in the intact spinotrapezius muscle. AB - The purpose of this study was to determine whether short-term exposure to clinically relevant concentrations of Pseudomonas aeruginosa lipopolysaccharide (LPS) impairs vasoreactivity of resistance arterioles in the intact spinotrapezius muscle microcirculation and, if so, to determine the mechanisms mediating this response. Using intravital microscopy, we found that 60-min suffusion of P. aeruginosa LPS (0.03-3.0 microg/ml) on the in situ hamster spinotrapezius muscle elicited an immediate, profound, and prolonged concentration-dependent vasodilation (P < 0.05). This response was reversible once suffusion of P. aeruginosa LPS was stopped. Pretreatment with N(G)-nitro-L arginine methyl ester (10.0 microM), a nonselective nitric oxide (NO) synthase inhibitor, but not N(G)-nitro-D-arginine methyl ester, abrogated P. aeruginosa LPS-induced vasodilation and elicited a small, albeit significant, vasoconstriction. Indomethacin had no significant effects on P. aeruginosa LPS induced responses. P. aeruginosa LPS had no significant effects on acetylcholine- and nitroglycerin-induced vasodilation in the spinotrapezius muscle. Collectively, these data indicate that short-term exposure to clinically relevant concentrations of P. aeruginosa LPS evokes an immediate, potent, prolonged, and reversible NO-dependent, prostaglandin-independent vasodilation in skeletal muscles in vivo. We suggest this response could play an important role in the pathophysiology of the profound vasomotor dysfunction observed in the peripheral circulation of patients with P. aeruginosa sepsis syndrome. PMID- 11408452 TI - Gender differences in leucine, but not lysine, kinetics. AB - There is a controversy in the literature as to the effects of gender on leucine kinetics. Two research groups found that men oxidize more leucine during exercise, whereas another group showed no gender effects. The purpose of our study was to examine the effects of gender on leucine and, for comparison purposes, lysine kinetics. Our subjects (n = 14) were seven matched pairs of men and women selected for their exercise habits and age. After 1 wk of a standardized diet, they exercised at 50% of maximal O(2) uptake for 1 h. There was an effect of exercise in both genders: an increased leucine oxidation and an attenuation in nonoxidative leucine disposal compared with rest (P < 0.05). Furthermore, our study confirms that there are gender differences in leucine, but not lysine, kinetics. Men had a higher rate of leucine oxidation and a lower rate of nonoxidative leucine disposal during exercise (P < 0.05). For women, a larger proportion of their exercise energy needs came from fat; for men, a greater fraction came from carbohydrate (P < 0.05). We conclude that female exercisers rely to a greater extent on fat as an energy source, thereby using less carbohydrate, amino acid, and protein as a fuel source. PMID- 11408453 TI - Flow-mediated release of nitric oxide in isolated, perfused rabbit lungs. AB - The effects of changing perfusate flow on lung nitric oxide (NO) production and pulmonary arterial pressure (Ppa) were tested during normoxia and hypoxia and after N(G)-monomethyl-L-arginine (L-NMMA) treatment during normoxia in both blood and buffer-perfused rabbit lungs. Exhaled NO (eNO) was unaltered by changing perfusate flow in blood-perfused lungs. In buffer-perfused lungs, bolus injections of ACh into the pulmonary artery evoked a transient increase in eNO from 67 +/- 3 (SE) to 83 +/- 7 parts/billion with decrease in Ppa, whereas perfusate NO metabolites (pNOx) remained unchanged. Stepwise increments in flow from 25 to 150 ml/min caused corresponding stepwise elevations in eNO production (46 +/- 2 to 73 +/- 3 nl/min) without changes in pNOx during normoxia. Despite a reduction in the baseline level of eNO, flow-dependent increases in eNO were still observed during hypoxia. L-NMMA caused declines in both eNO and pNOx with a rise in Ppa. Pulmonary vascular conductance progressively increased with increasing flow during normoxia and hypoxia. However, L-NMMA blocked the flow dependent increase in conductance over the range of 50-150 ml/min of flow. In the more physiological conditions of blood perfusion, eNO does not reflect endothelial NO production. However, from the buffer perfusion study, we suggest that endothelial NO production secondary to increasing flow, may contribute to capillary recruitment and/or shear stress-induced vasodilation. PMID- 11408454 TI - Role of intrinsic airway neurons in ozone-induced airway hyperresponsiveness in ferret trachea. AB - Exposure to ozone (O(3)) enhances airway responsiveness, which is mediated partly by the release of substance P (SP) from airway neurons. In this study, the role of intrinsic airway neurons in O(3)-induced airway responses was examined. Ferrets were exposed to 2 ppm O(3) or air for 1 h. Reactivity of isolated tracheal smooth muscle to cholinergic agonists was significantly increased after O(3) exposure, as were contractions to electrical field stimulation at 10 Hz. Pretreatment with CP-99994, a neurokinin type 1 receptor antagonist, partially abolished the O(3)-induced reactivity to cholinergic agonists and electrical field stimulation. The O(3)-enhanced airway responses were present in tracheal segments cultured for 24 h, a procedure shown to deplete sensory nerves while maintaining viability of intrinsic airway neurons, and all the enhanced smooth muscle responses were also diminished by CP-99994. Immunocytochemistry showed that the percentage of SP-containing neurons in longitudinal trunk and the percentage of neurons innervated by SP-positive nerve fibers in superficial muscular plexus were significantly increased at 1 h after exposure to O(3). These results suggest that enhanced SP levels in airway ganglia contribute to O(3) induced airway hyperresponsiveness. PMID- 11408455 TI - Expression of heme oxygenase in the oxygen-sensing regions of the rostral ventrolateral medulla. AB - Recently, unique regions in the rostral ventrolateral medulla (RVLM) have been found to be oxygen sensitive. However, the mechanism of sensing oxygen in these RVLM regions is unknown. Because heme oxygenase (HO) has been shown to be involved in the hypoxic responses of the carotid body and pulmonary artery, the aim of this study was to determine whether HO is present in the RVLM and whether expression of HO is altered by chronic hypoxia. Adult rats were exposed to hypoxia (10% O(2)) or normoxia (21% O(2)) for 10 days, and the mRNA for HO-1 and HO-2 was examined in the RVLM by using RT-PCR. Expression of HO-2 mRNA was seen in the RVLM of both control and hypoxic samples, whereas expression of HO-1 mRNA was only seen in the RVLM of hypoxic samples. HO-2 was immunocytochemically localized in brain sections (40 microm) to the C1 region and pre-Botzinger complex of the RVLM. Together, these results indicate that HO-2 is present in the RVLM under control conditions and that HO-1 is induced in the RVLM during chronic hypoxia, consistent with a potential role for HO in the oxygen-sensing function of these cardiorespiratory RVLM regions. PMID- 11408456 TI - Validity of estimating limb muscle volume by bioelectrical impedance. AB - The present study aimed to investigate the validity of estimating muscle volume by bioelectrical impedance analysis. Bioelectrical impedance and series cross sectional images of the forearm, upper arm, lower leg, and thigh on the right side were determined in 22 healthy young adult men using a specially designed bioelectrical impedance acquisition system and magnetic resonance imaging (MRI) method, respectively. The impedance index (L(2)/Z) for every segment, calculated as the ratio of segment length squared to the impedance, was significantly correlated to the muscle volume measured by MRI, with r = 0.902-0.976 (P < 0.05). In these relationships, the SE of estimation was 38.4 cm(3) for the forearm, 40.9 cm(3) for the upper arm, 107.2 cm(3) for the lower leg, and 362.3 cm(3) for the thigh. Moreover, isometric torque developed in elbow flexion or extension and knee flexion or extension was significantly correlated to the L(2)/Z values of the upper arm and thigh, respectively, with correlation coefficients of 0.770 0.937 (P < 0.05), which differed insignificantly from those (0.799-0.958; P < 0.05) in the corresponding relationships with the muscle volume measured by MRI of elbow flexors or extensors and knee flexors or extensors. Thus the present study indicates that bioelectrical impedance analysis may be useful to predict the muscle volume and to investigate possible relations between muscle size and strength capability in a limited segment of the upper and lower limbs. PMID- 11408457 TI - Respiratory abdominal muscle recruitment and chest wall motion in myotonic muscular dystrophy. AB - Abdominal muscles are selectively active in normal subjects during stress and may increase the potential energy for inspiration by reducing the end-expiratory lung volume (EELV). We hypothesized that a similar process would occur in subjects with myotonic muscular dystrophy (MMD), but would be less effective, because of to their weakness and altered chest wall mechanics. Fine-wire electromyography (EMG) of the transversus abdominis (TA), internal oblique (IO), external oblique, and rectus abdominis was recorded in 10 MMD and 10 control subjects. EMG activity, respiratory inductive plethysmography, and gastric pressure were recorded during static pressure measurement and at increasing levels of inspiratory resistance breathing. EELV was reduced and chest wall motion was synchronous only in controls. Although the TA and IO were selectively recruited in both groups, EMG activity of the MMD group was twice that of controls at the same inspiratory pressure. In MMD subjects with mildly reduced forced vital capacity, significant differences can be seen in abdominal muscle recruitment, wall motion, work of breathing, and ventilatory parameters. PMID- 11408458 TI - Dynamic ventilatory response to CO(2) in congestive heart failure patients with and without central sleep apnea. AB - Nonobstructive (i.e., central) sleep apnea is a major cause of sleep-disordered breathing in patients with stable congestive heart failure (CHF). Although central sleep apnea (CSA) is prevalent in this population, occurring in 40-50% of patients, its pathogenesis is poorly understood. Dynamic loop gain and delay of the chemoreflex response to CO(2) was measured during wakefulness in CHF patients with and without CSA by use of a pseudorandom binary CO(2) stimulus method. Use of a hyperoxic background minimized responses derived from peripheral chemoreceptors. The closed-loop and open-loop gain, estimated from the impulse response, was three times greater in patients with nocturnal CSA (n = 9) than in non-CSA patients (n = 9). Loop dynamics, estimated by the 95% response duration time, did not differ between the two groups of patients. We speculate that an increase in dynamic gain of the central chemoreflex response to CO(2) contributes to the genesis of CSA in patients with CHF. PMID- 11408459 TI - Maximal and submaximal forces of slow fibers in human soleus after bed rest. AB - The effects of 2 and 4 mo of bed rest, with or without exercise countermeasures, on the contractile properties of slow fibers in the human soleus muscle were examined. Mean fiber diameters were 8 and 36% smaller after 2 and 4 mo of bed rest, respectively, than the pre-bed rest level. Maximum tetanic force (P(o)), maximum activated force (F(max)) per cross-sectional area (CSA), and the common logarithm value of free Ca(2+) concentration required for half-maximal activation (pCa(50)) also decreased after 2 and 4 mo of bed rest. In contrast, maximum unloaded shortening velocity (V(o)) was increased after 2 and 4 mo of bed rest. After 1 mo of recovery, fiber diameters, P(o), F(max) per CSA (P > 0.05), and pCa(50) were increased and V(o) decreased toward pre-bed rest levels. Effects of knee extension/flexion exercise by wearing an anti-G Penguin suit for 10 h daily, and the effects of loading or unloading of the plantar flexors with (Penguin-1) or without (Penguin-2) placing the elastic loading elements of the suit, respectively, were investigated during ~2 mo of bed rest. In the Penguin-1 group, mean fiber diameter, P(o), F(max) per CSA, V(o), and pCa(50) were similar before and after bed rest. However, the responses of fiber size and contractile properties to bed rest were not prevented in the Penguin-2 group, although the degree of the changes was less than those induced by bed rest without any countermeasure. These results indicate that long-term bed rest results in reductions of fiber size, force-generation capacity, and Ca(2+) sensitivity, and enhancement of shortening velocity in slow fibers of the soleus. The data indicate that continuous mechanical loading on muscle, such as stretching of muscle, is an effective countermeasure for the prevention of muscular adaptations to gravitational unloading. PMID- 11408460 TI - Leukocyte counts and lymphocyte responsiveness associated with repeated bouts of strenuous endurance exercise. AB - This study compared leukocyte counts and lymphocyte responsiveness during and after a second bout of high-intensity endurance exercise on the same day with the response to a similar but single bout of exercise. Nine athletes participated in three 24-h trials: 1) rest in bed (Rest); 2) one bout of exercise (One); and 3) two bouts of exercise (Two). All bouts consisted of 75 min at approximately 75% of maximal O(2) uptake on a cycle ergometer. Lymphocytes in whole blood were stimulated with monoclonal antibodies against CD2 and assessed by flow cytometry for expression of the early activation molecule CD69. The second bout of exercise in the Two trial was associated with significantly increased concentrations of total leukocytes, neutrophils, lymphocytes, CD4(+), CD8(+), and CD56(+) cells and a significantly decreased percentage of CD56(+) cells expressing CD69 compared with a single bout. Additionally, there was a significantly decreased CD69 fluorescence in CD56(+) cells postexercise. These differences suggest a "carry over" effect in the immune system from a first to a second bout of exercise on the same day. PMID- 11408461 TI - Impaired distensibility of the left ventricle after stiffening of the right ventricle. AB - Acute and chronic alterations of right ventricular (RV) wall properties can change left ventricular (LV) performance. We investigated whether and how stiffening of the RV free wall alters LV diastolic distensibility. We used cross circulated isolated hearts, in which the LV and RV were independently controllable. Stiffness of the RV free wall was altered by intramuscular injections of glutaraldehyde into the RV free wall after right coronary artery ligation. We measured circumferential and longitudinal regional lengths in the septum and LV free wall. During data acquisition, RV volume was held constant. After the RV free wall was stiffened by glutaraldehyde, the LV diastolic pressure volume relation shifted upward and became steeper. Importantly, stiffening of the RV free wall increased the diastolic regional area in the septum and LV free wall under constant LV volume. The augmented regional dimensions may result in enhanced regional tension under constant LV volume and may be related to the observed increase in LV diastolic intracavitary pressure. The impaired LV diastolic distensibility by stiffening of the RV free wall may be at least partly explained by myocardial stretch, probably due to LV deformation. PMID- 11408462 TI - Lower inflection point and recruitment with PEEP in ventilated patients with acute respiratory failure. AB - The lower inflection point (LIP) on the total respiratory system pressure-volume (P-V) curve is widely used to set positive end-expiratory pressure (PEEP) in patients with acute respiratory failure (ARF) on the assumption that LIP represents alveolar recruitment. The aims of this work were to study the relationship between LIP and recruited volume (RV) and to propose a simple method to quantify the RV. In 23 patients with ARF, respiratory system P-V curves were obtained by means of both constant-flow and rapid occlusion technique at four different levels of PEEP and were superimposed on the same P-V plot. The RV was measured as the volume difference at a pressure of 20 cm H(2)O. A third measurement of the RV was done by comparing the exhaled volumes after the same distending pressure of 20 cm H(2)O was applied (equal pressure method). RV increased with PEEP (P < 0.0001); the equal pressure method compares favorably with the other methods (P = 0.0001 by correlation), although individual data cannot be superimposed. No significant difference was found when RV was compared with PEEP in the group of patients with a LIP < or =5 cm H(2)O and the group with a LIP >5 cm H(2)O (76.9 +/- 94.3 vs. 61.2 +/- 51.3, 267.7 +/- 109.9 vs. 209.6 +/- 73.9, and 428.2 +/- 216.3 vs. 375.8 +/- 145.3 ml with PEEP of 5, 10, and 15 cm H(2)O, respectively). A RV was found even when a LIP was not present. We conclude that the recruitment phenomenon is not closely related to the presence of a LIP and that a simple method can be used to measure RV. PMID- 11408463 TI - Ambulatory estimates of maximal aerobic power from foot -ground contact times and heart rates in running humans. AB - Seeking to develop a simple ambulatory test of maximal aerobic power (VO(2 max)), we hypothesized that the ratio of inverse foot-ground contact time (1/t(c)) to heart rate (HR) during steady-speed running would accurately predict VO(2 max). Given the direct relationship between 1/t(c) and mass-specific O(2) uptake during running, the ratio 1/t(c). HR should reflect mass-specific O(2) pulse and, in turn, aerobic power. We divided 36 volunteers into matched experimental and validation groups. VO(2 max) was determined by a treadmill test to volitional fatigue. Ambulatory monitors on the shoe and chest recorded foot-ground contact time (t(c)) and steady-state HR, respectively, at a series of submaximal running speeds. In the experimental group, aerobic fitness index (1/t(c). HR) was nearly constant across running speed and correlated with VO(2 max) (r = 0.90). The regression equation derived from data from the experimental group predicted VO(2 max) from the 1/t(c). HR values in the validation group within 8.3% and 4.7 ml O(2) x kg(-1) x min(-1) (r = 0.84) of measured values. We conclude that simultaneous measurements of foot-ground constant times and heart rates during level running at a freely chosen constant speed can provide accurate estimates of maximal aerobic power. PMID- 11408464 TI - alpha-Actin: disposition, quantities, and estimated effects on lung recoil and compliance. AB - We have investigated the basis and implications of pneumoconstriction by measuring disposition and quantities of alpha-smooth muscle actin in rat and guinea pig lungs and modeling its effects on lung recoil and compliance. A robust marker of contractility, alpha-smooth muscle actin appears in smooth muscle or myofibroblast-like cells in pleura, airways, blood vessels, and alveolar ductal tissues. In each site, we measured its transected area by immunofluorescent staining and frequency-modulated scanning confocal microscopy. We incorporated these data in a model of the parenchyma consisting of an extensive elastic network with embedded contractile structures. We conclude that contraction at any one of these sites alone can decrease parenchymal compliance by 20-30% during tidal breathing. This is due mostly to the stiffness of activated contractile elements undergoing passive cycling; constant muscle tension would have little effect. The magnitude of the effect corresponds with known responses of the lung to hypocapnia, consistent with a homeostatic function in which gas exchange is defended by redistributing ventilation away from overventilated units. PMID- 11408465 TI - Exhaled NO: first, hold your breath. PMID- 11408466 TI - A single-breath technique with variable flow rate to characterize nitric oxide exchange dynamics in the lungs. AB - Current techniques to estimate nitric oxide (NO) production and elimination in the lungs are inherently nonspecific or are cumbersome to perform (multiple breathing maneuvers). We present a new technique capable of estimating key flow independent parameters characteristic of NO exchange in the lungs: 1) the steady state alveolar concentration (C(alv,ss)), 2) the maximum flux of NO from the airways (J(NO,max)), and 3) the diffusing capacity of NO in the airways (D(NO,air)). Importantly, the parameters were estimated from a single experimental single-exhalation maneuver that consisted of a preexpiratory breath hold, followed by an exhalation in which the flow rate progressively decreased. The mean values for J(NO,max), D(NO,air), and C(alv,ss) do not depend on breath hold time and range from 280-600 pl/s, 3.7-7.1 pl. s(-1). parts per billion (ppb)(-1), and 0.73-2.2 ppb, respectively, in two healthy human subjects. A priori estimates of the parameter confidence intervals demonstrate that a breath hold no longer than 20 s may be adequate and that J(NO,max) can be estimated with the smallest uncertainty and D(NO,air) with the largest, which is consistent with theoretical predictions. We conclude that our new technique can be used to characterize flow-independent NO exchange parameters from a single experimental single-exhalation breathing maneuver. PMID- 11408467 TI - Invited review: significance of spatial and temporal heterogeneity of calcium transients in smooth muscle. AB - The multiplicity of mechanisms involved in regulation of intracellular Ca(2+) concentration ([Ca(2+)](i)) in smooth muscle results in both intra- and intercellular heterogeneities in [Ca(2+)](i). Heterogeneity in [Ca(2+)](i) regulation is reflected by the presence of spontaneous, localized [Ca(2+)](i) transients (Ca(2+) sparks) representing Ca(2+) release through ryanodine receptor (RyR) channels. Ca(2+) sparks display variable spatial Ca(2+) distributions with every occurrence within and across cellular regions. Individual sparks are often grouped, and fusion of sparks produces large local elevations in [Ca(2+)](i) that occasionally trigger propagating [Ca(2+)](i) waves. Ca(2+) sparks may modulate membrane potential and thus smooth muscle contractility. Sparks may also be the target of other regulatory factors in smooth muscle. Agonists induce propagating [Ca(2+)](i) oscillations that originate from foci with high spark incidence and also represent Ca(2+) release through RyR channels. With increasing agonist concentration, the peak of regional [Ca(2+)](i) oscillations remains relatively constant, whereas both frequency and propagation velocity increase. In contrast, the global cellular response appears as a concentration-dependent increase in peak as well as mean cellular [Ca(2+)](i), representing a spatial and temporal integration of the oscillations. The significance of agonist-induced [Ca(2+)](i) oscillations lies in the establishment of a global [Ca(2+)](i) level for slower Ca(2+)-dependent physiological processes. PMID- 11408468 TI - Invited review: regulation of myosin phosphorylation in smooth muscle. AB - Phosphorylation of the regulatory light chains of myosin II (rMLC) by the Ca(2+)/calmodulin-dependent myosin light-chain kinase (MLCK) and dephosphorylation by a type 1 phosphatase (MLCP), which is targeted to myosin by a regulatory subunit (MYPT1), are the predominant mechanisms of regulation of smooth muscle tone. The activities of both enzymes are modulated by several protein kinases. MLCK is inhibited by the Ca(2+)/calmodulin-dependent protein kinase II, whereas the activity of MLCP is increased by cGMP and perhaps also cAMP-dependent protein kinases. In either case, this results in a decrease in the Ca(2+) sensitivity of rMLC phosphorylation and force production. The activity of MLCP is inhibited by Rho-associated kinase, one of the effectors of the monomeric GTPase Rho, and protein kinase C, leading to an increase in Ca(2+) sensitivity. Hence, smooth muscle tone appears to be regulated by a network of activating and inactivating intracellular signaling cascades. PMID- 11408470 TI - Selected contribution: airway caliber in healthy and asthmatic subjects: effects of bronchial challenge and deep inspirations. AB - In 9 healthy and 14 asthmatic subjects before and after a standard bronchial challenge and a modified [deep inspiration (DI), inhibited] bronchial challenge and after albuterol, we tracked airway caliber by synthesizing a method to measure airway resistance (Raw; i.e., lung resistance at 8 Hz) in real time. We determined the minimum Raw achievable during a DI to total lung capacity and the subsequent dynamics of Raw after exhalation and resumption of tidal breathing. Results showed that even after a bronchial challenge healthy subjects can dilate airways maximally, and the dilation caused by a single DI takes several breaths to return to baseline. In contrast, at baseline, asthmatic subjects cannot maximally dilate their airways, and this worsens considerably postconstriction. Moreover, after a DI, the dilation that does occur in airway caliber in asthmatic subjects constricts back to baseline much faster (often after a single breath). After albuterol, asthmatic subjects could dilate airways much closer to levels of those of healthy subjects. These data suggest that the asthmatic smooth muscle resides in a stiffer biological state compared with the stimulated healthy smooth muscle, and inhibiting a DI in healthy subjects cannot mimic this. PMID- 11408471 TI - Selected contribution: effect of volatile anesthetics on cADP-ribose-induced Ca(2+) release system. AB - Volatile anesthetics have multiple actions on intracellular Ca(2+) homeostasis, including activation of the ryanodine channel (RyR) and sensitization of this channel to agonists such as caffeine and ryanodine. Recently it has been described that the nucleotide cADP-ribose (cADPR) is the endogenous regulator of the RyR in many mammalian cells, and cADPR has been proposed to be a second messenger in many signaling pathways. I investigated the effect of volatile anesthetics on the cADPR signaling system, using sea urchin egg homogenates as a model of intracellular Ca(2+) stores. Ca(2+) uptake and release were monitored in sea urchin egg homogenates by using the fluo-3 fluorescence technique. Activity of the ADP-ribosyl cyclase was monitored by using a fluorometric method using nicotinamide guanine dinucleotide as a substrate. Halothane in concentrations up to 800 microM did not induce Ca(2+) release by itself in sea urchin egg homogenates. However, halothane potentiates the Ca(2+) release mediated by agonists of the ryanodine channel, such as ryanodine. Furthermore, other volatile anesthetics such as isoflurane and sevoflurane had no effect. Halothane also potentiated the activation of the ryanodine channel mediated by the endogenous nucleotide cADPR. The half-maximal concentration for cADPR-induced Ca(2+) release was decreased about three times by addition of 800 microM halothane. The reverse was also true: addition of subthreshold concentrations of cADPR sensitized the homogenates to halothane. In contrast, all the volatile anesthetics used had no effect on the activity of the enzyme that synthesizes cADPR. I propose that the complex effect of volatile anesthetics on intracellular Ca(2+) homeostasis may involve modulation of the cADPR signaling system. PMID- 11408472 TI - Selected contribution: NO released to flow reduces myogenic tone of skeletal muscle arterioles by decreasing smooth muscle Ca(2+) sensitivity. AB - To clarify the contribution of intracellular Ca(2+) concentration ([Ca(2+)](i)) dependent and -independent signaling mechanisms in arteriolar smooth muscle (aSM) to modulation of arteriolar myogenic tone by nitric oxide (NO), released in response to increases in intraluminal flow from the endothelium, changes in aSM [Ca(2+)](i) and diameter of isolated rat gracilis muscle arterioles (pretreated with indomethacin) were studied by fluorescent videomicroscopy. At an intraluminal pressure of 80 mmHg, [Ca(2+)](i) significantly increased and myogenic tone developed in response to elevations of extracellular Ca(2+) concentration. The Ca(2+) channel inhibitor nimodipine substantially decreased [Ca(2+)](i) and completely inhibited myogenic tone. Dilations to intraluminal flow (that were inhibited by N(omega)-nitro-L-arginine methyl ester) or dilations to the NO donor S-nitroso-N-acetyl-DL-penicillamine (that were inhibited by the guanylate cyclase inhibitor 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one) were not accompanied by substantial decreases in aSM [Ca(2+)](i). 8-Bromoguanosine cGMP and the cGMP-specific phosphodiesterase inhibitor zaprinast significantly dilated arterioles yet elicited only minimal decreases in [Ca(2+)](i). Thus flow-induced endothelial release of NO elicits relaxation of arteriolar smooth muscle by a cGMP-dependent decrease of the Ca(2+) sensitivity of the contractile apparatus without substantial changes in the pressure-induced level of [Ca(2+)](i). PMID- 11408473 TI - An enhanced mutant of red fluorescent protein DsRed for double labeling and developmental timer of neural fiber bundle formation. AB - We developed a new variant of coral-derived red fluorescent protein, DsRed S197Y, which is brighter and essentially free from secondary fluorescence peak. This makes it an ideal reporter for double labeling with green fluorescent protein (GFP). Though purified protein shows only 20% stronger fluorescence emission, culture cells that express DsRed S197Y exhibit a 3-3.5 times higher level of fluorescence than the cells that express wild-type DsRed. The much slower fluorescence maturation of DsRed than that of GFP is a beneficial feature for a fluorescent developmental timer application. When GFP and DsRed S197Y are expressed simultaneously, emissions start at different latency. This provides information about the time after the onset of expression. It reflects the order of cell differentiation if the expression is activated upon differentiation of certain types of cells. We applied this system to the developing brain of Drosophila and visualized, for the first time, the formation order of neural fibers within a large bundle. Our results showed that newly extending fibers of the mushroom body neurons mainly run into the core rather than to the periphery of the existing bundle. DsRed-based timer thus presents an indispensable tool for developmental biology and genetics of model organisms. PMID- 11408474 TI - Eukaryotic initiation factors 4A (eIF4A) and 4G (eIF4G) mutually interact in a 1:1 ratio in vivo. AB - mRNA translation in eukaryotic cells involves a set of proteins termed translation initiation factors (eIFs), several of which are involved in the binding of ribosomes to mRNA. These include eIF4G, a modular scaffolding protein, and eIF4A, an RNA helicase, of which two closely related forms are known in mammals, eIF4A(I) and eIF4A(II). In mammals, eIF4G possesses two independent sites for binding eIF4A, whereas in other eukaryotes (e.g. yeast) only one site appears to be present, thus raising the issue of the stoichiometry of eIF4G.eIF4A complexes in different eukaryotes. We show that in human embryonic kidney cells eIF4G is associated with eIF4A(I) or eIF4A(II) but not with both simultaneously, suggesting a stoichiometry of 1:1 rather than 1:2. To confirm this, eIF4A(I) or eIF4A(II) was expressed in a tagged form in these cells, and complexes with eIF4G were again isolated. Complexes containing tagged eIF4A(I) or eIF4A(II) contained no endogenous eIF4A, supporting the notion that eIF4G binds only one molecule of eIF4A. Each binding site in eIF4G can bind either eIF4A(I) or eIF4A(II). The data imply that the second binding site in mammalian eIF4A does not bind an additional eIF4A molecule and that initiation factor complexes in different eukaryotes contain one eIF4A per eIF4G. PMID- 11408475 TI - Notch1 and amyloid precursor protein are competitive substrates for presenilin1 dependent gamma-secretase cleavage. AB - Proteolytic processing of the amyloid precursor protein (APP) by beta- and gamma secretases results in the production of a highly amyloidogenic Abeta peptide, which deposits in the brains of Alzheimer's disease patients. Similar gamma secretase processing occurs in another transmembrane protein, Notch1, releasing a potent signaling molecule, the Notch C-terminal domain. It has been shown that both events are dependent on a presenilin-dependent protease. We now test the hypothesis that activated Notch1 and APP are competitive substrates for the same proteolytic activity in neurons. Treatment of neurons with the native Notch ligand, Delta, induces endogenous Notch1 intramembraneous cleavage and diminishes Abeta production in a dose-dependent manner. Complementary experiments showed that the converse was also true. Overexpressing human APP (APP(695Sw)) in neurons leads to a decrease in endogenous Notch1 signal transduction, as assessed by a CBF1 luciferase transcription assay, by Notch C-terminal domain nuclear translocation in vitro and by analysis of Notch C-terminal domain generation and Notch1 staining in vivo. In summary, two complementary approaches suggest that APP and Notch1 are physiologically relevant competitive substrates for gamma secretase activity. PMID- 11408476 TI - TUCAN, an antiapoptotic caspase-associated recruitment domain family protein overexpressed in cancer. AB - Caspase-associated recruitment domains (CARDs) are protein interaction domains that participate in activation or suppression of CARD-carrying members of the caspase family of apoptosis-inducing proteases. A novel CARD-containing protein was identified that is overexpressed in some types of cancer and that binds and suppresses activation of procaspase-9, which we term TUCAN (tumor-up-regulated CARD-containing antagonist of caspase nine). The CARD domain of TUCAN selectively binds itself and procaspase-9. TUCAN interferes with binding of Apaf1 to procaspase-9 and suppresses caspase activation induced by the Apaf1 activator, cytochrome c. Overexpression of TUCAN in cells by stable or transient transfection inhibits apoptosis and caspase activation induced by Apaf1/caspase-9 dependent stimuli, including Bax, VP16, and staurosporine, but not by Apaf1/caspase-9-independent stimuli, Fas and granzyme B. High levels of endogenous TUCAN protein were detected in several tumor cell lines and in colon cancer specimens, correlating with shorter patient survival. Thus, TUCAN represents a new member of the CARD family that selectively suppresses apoptosis induced via the mitochondrial pathway for caspase activation. PMID- 11408477 TI - Bone morphogenetic protein-2 inhibits serum deprivation-induced apoptosis of neonatal cardiac myocytes through activation of the Smad1 pathway. AB - Bone morphogenetic protein (BMP)-2 has been shown to induce ectopic expression of cardiac transcription factors and beating cardiomyocytes in non-precardiac mesodermal cells, suggesting that BMP-2 is an inductive signaling molecule that participates in cardiac development. However, direct evidence of the effects of BMP-2 on cardiac myocytes has not been reported. To examine the role of BMP-2 and its receptors, we studied the ability of BMP-2 to promote survival of isolated neonatal rat cardiac myocytes. BMP receptors IA, IB, and II and activin receptor I were found to be expressed in myocytes, and BMP-2 phosphorylated Smad1 and p38 MAPK. Interestingly, BMP-2 promoted survival and inhibited apoptosis of serum deprived myocytes, although it did not strongly induce hypertrophic growth. To explore the mechanisms for this protective effect, an adenovirus-based vector system was used. Similar to BMP-2, Smad1 promoted survival that was repressed by Smad6. Moreover, BMP-2 and Smad1 enhanced the expression of the anti-apoptotic molecule Bcl-x(L). Antisense oligonucleotides to bcl-x(L) attenuated the survival effected by BMP-2. Overall, our findings suggest that BMP-2 prevents apoptosis of myocytes by induction of Bcl-x(L) via a Smad1 pathway and might be a novel survival factor without any hypertrophic effect on myocytes. PMID- 11408478 TI - Kinetics of beta-lactam interactions with penicillin-susceptible and -resistant penicillin-binding protein 2x proteins from Streptococcus pneumoniae. Involvement of acylation and deacylation in beta-lactam resistance. AB - Kinetic interactions of beta-lactam antibiotics such as penicillin-G and cefotaxime with normal, penicillin-susceptible PBP2x from Streptococcus pneumoniae and a penicillin-resistant PBP2x (PBP2x(R)) from a resistant clinical isolate (CS109) of the bacterium have been extensively characterized using electrospray mass spectrometry coupled with a fast reaction (quench flow) technique. Kinetic evidence for a two-step acylation of PBP2x by penicillin-G has been demonstrated, and the dissociation constant, K(d) of 0.9 mm, and the acylation rate constant, k(2) of 180 s(-1), have been determined for the first time. The millimolar range K(d) implies that the beta-lactam fits to the active site pocket of the penicillin-sensitive PBP rather poorly, whereas the extremely fast k(2) value indicates that this step contributes most of the binding affinity of the beta-lactam. The values of K(d) (4 mm) and k(2) (0.56 s(-1)) were also determined for PBP2x(R). The combined value of k(2)/K(d), known as overall binding efficiency, for PBP2x(R) (137 m(-1) s(-1)) was over 1000-fold slower than that for PBP2x (200,000 m(-1) s(-1)), indicating that a major part is played by the acylation steps in penicillin resistance. Most of the decreased binding efficiency of PBP2x(R) comes from the decreased ( approximately 300-fold) k(2). Kinetic studies of cefotaxime acylation of the two PBP2x proteins confirmed all of the above findings. Deacylation rate constants (k(3)) for the third step of the interactions were determined to be 8 x 10(-6) s(-1) for penicilloyl-PBP2x and 5.7 x 10(-4) s(-1) for penicilloyl-PBP2x(R), corresponding to over 70-fold increase of the deacylation rate for the resistant PBP2x(R). Similarly, over 80 fold enhancement of the deacylation rate was found for cefotaxime-PBP2x(R) complex (k(3) = 3 x 10(-4) s(-1)) as compared with that of cefotaxime-PBP2x complex (3.5 x 10(-6) s(-1)). This is the first time that such a significant increase of k(3) values was found for a beta-lactam-resistant penicillin-binding protein. These data indicate that the deacylation step also plays a role, which is much more important than previously thought, in PBP2x(R) resistance to beta lactams. PMID- 11408479 TI - An in vitro reducing system for the enzymic conversion of cobalamin to adenosylcobalamin. AB - Homogeneous ferredoxin (flavodoxin):NADP(+) reductase and flavodoxin A proteins served as electron donors for the reduction of co(III)rrinoids to co(I)rrinoids in vitro. The resulting co(I)rrinoids served as substrates for the ATP:co(I)rrinoid adenosyltransferase (CobA) enzyme of Salmonella enterica serovar Typhimurium LT2 and were converted to their respective adenosylated derivatives. The reaction products were isolated by reverse phase high performance liquid chromatography, and their identities were confirmed by UV-visible spectroscopy, mass spectrometry, and in vivo biological activity assays. Adenosylcobalamin generated by this system supported the activity of 1,2-propanediol dehydratase as effectively as authentic adenosylcobalamin. This is the first report of a protein system that can be coupled to the adenosyltransferase CobA enzyme for the conversion of co(III)rrinoids to their adenosylated derivatives. PMID- 11408480 TI - Amino acid residues that are critical for in vivo catalytic activity of CtpA, the carboxyl-terminal processing protease for the D1 protein of photosystem II. AB - CtpA, a carboxyl-terminal processing protease, is a member of a novel family of endoproteases that includes a tail-specific protease from Escherichia coli. In oxygenic photosynthetic organisms, CtpA catalyzes C-terminal processing of the D1 protein of photosystem II, an essential event for the assembly of a manganese cluster and consequent light-mediated water oxidation. We introduced site specific mutations at 14 conserved residues of CtpA in the cyanobacterium Synechocystis sp. PCC 6803 to examine their functional roles. Analysis of the photoautotrophic growth capabilities of these mutants, their ability to process precursor D1 protein and hence evolve oxygen, along with an estimation of the protease content in the mutants revealed that five of these residues are critical for in vivo activity of CtpA. Recent x-ray crystal structure analysis of CtpA from the eukaryotic alga Scenedesmus obliquus (Liao, D.-I., Qian, J., Chisholm, D. A., Jordan, D. B. and Diner, B. A. (2000) Nat. Struct. Biol. 7, 749-753) has shown that the residues equivalent to Ser-313 and Lys-338, two of the five residues mentioned above, form the catalytic center of this enzyme. Our in vivo analysis demonstrates that the three other residues, Asp-253, Arg-255, and Glu 316, are also important determinants of the catalytic activity of CtpA. PMID- 11408481 TI - The protein kinase Gcn2p mediates sodium toxicity in yeast. AB - Phosphorylation of the alpha-subunit of eukaryotic initiation factor 2 (eIF2alpha) is a conserved mechanism regulating protein synthesis in response to various stresses. A screening for negative factors in yeast salt stress tolerance has led to the identification of Gcn2p, the single yeast eIF2alpha kinase that is activated by amino acid starvation in the general amino acid control response. Mutation of other components of this regulatory circuit such as GCN1 and GCN3 also resulted in improved NaCl tolerance. The gcn2 phenotype was not accompanied by changes in sodium or potassium homeostasis. NaCl induced a Gcn2p-dependent phosphorylation of eIF2alpha and translational activation of Gcn4p, the transcription factor that mediates the general amino acid control response. Mutations that activate Gcn4p function, such as gcd7-201, cpc2, and deletion of the translational regulatory region of the GCN4 gene, also cause salt sensitivity. It can be postulated that sodium activation of the Gcn2p pathway has toxic effects on growth under NaCl stress and that this novel mechanism of sodium toxicity may be of general significance in eukaryotes. PMID- 11408482 TI - Procollagen II amino propeptide processing by ADAMTS-3. Insights on dermatosparaxis. AB - The amino and carboxyl propeptides of procollagens I and II are removed by specific enzymes as a prerequisite for fibril assembly. Null mutations in procollagen I N-propeptidase (ADAMTS-2) cause dermatosparaxis in cattle and the Ehlers-Danlos syndrome (dermatosparactic type) in humans by preventing proteolytic excision of the N-propeptide of procollagen I. We have found that procollagen II is processed normally in dermatosparactic nasal cartilage, suggesting the existence of another N-propeptidase(s). We investigated such a role for ADAMTS-3 in Swarm rat chondrosarcoma RCS-LTC cells, which fail to process the procollagen II N-propeptide. Stable transfection of RCS-LTC cells with bovine ADAMTS-2 or human ADAMTS-3 partially rescued the processing defect, suggesting that ADAMTS-3 has procollagen II N-propeptidase activity. Human skin and skin fibroblasts showed 30-fold higher mRNA levels of ADAMTS-2 than ADAMTS-3, whereas ADAMTS-3 mRNA was 5-fold higher than ADAMTS-2 mRNA in human cartilage. We propose that both ADAMTS-2 and ADAMTS-3 process procollagen II, but ADAMTS-3 is physiologically more relevant, given its preferred expression in cartilage. The findings provide an explanation for the sparing of cartilage in dermatosparaxis and, perhaps, for the relative sparing of some procollagen I-containing tissues. PMID- 11408483 TI - Binding and repair of mismatched DNA mediated by Rhp14, the fission yeast homologue of human XPA. AB - Rhp14 of Schizosaccharomyces pombe is homologous to human XPA and Saccharomyces cerevisiae Rad14, which act in nucleotide excision repair of DNA damages induced by ultraviolet light and chemical agents. Cells with disrupted rhp14 were highly sensitive to ultraviolet light, and epistasis analysis with swi10 (nucleotide excision repair) and rad2 (Uve1-dependent ultraviolet light damage repair pathway) revealed that Rhp14 is an important component of nucleotide excision repair for ultraviolet light-induced damages. Moreover, defective rhp14 caused instability of a GT repeat, similar to swi10 and synergistically with msh2 and exo1. Recombinant Rhp14 with an N-terminal hexahistidine tag was purified from Escherichia coli. Complementation studies with a rhp14 mutant demonstrated that the tagged Rhp14 is functional in repair of ultraviolet radiation-induced damages and in mitotic mutation avoidance. In bandshift assays, Rhp14 showed a preference to substrates with mismatched and unpaired nucleotides. Similarly, XPA bound more efficiently to C/C, A/C, and T/C mismatches than to homoduplex DNA. Our data show that mismatches and loops in DNA are substrates of nucleotide excision repair. Rhp14 is likely part of the recognition complex but alone is not sufficient for the high discrimination of nucleotide excision repair for modified DNA. PMID- 11408484 TI - A novel mouse beta-defensin, mBD-6, predominantly expressed in skeletal muscle. AB - Defensins comprise a family of cationic antimicrobial peptides that is characterized by the conserved 6 cysteine residues. They are expressed in the epithelial cells of various organs and are identified as key elements in the host defense system at the mucosal surface. We isolated a novel mouse beta-defensin gene from the bacterial artificial chromosome DNA containing the mouse beta defensin-3 gene. The full-length cDNA was cloned from skeletal muscle cDNA and called mouse beta-defensin-6 (mBD-6). The predicted peptide conserved the 6 cysteine motif and had 59% amino acid sequence identity with mouse beta-defensin 3 and 59% identity with mouse beta-defensin-4. We demonstrated the expression of mBD-6 in skeletal muscle in addition to the esophagus, tongue, and trachea. In animal models of endotoxemia, mBD-6 expression was also induced in the lung. mBD 6 showed potent antimicrobial activity against Escherichia coli and would play an important role in host defense in the esophagus, airways, and skeletal muscle. mBD-6 is the first reported beta-defensin predominantly expressed in skeletal muscle. This unique tissue specificity suggests some novel physiological roles of this peptide family. PMID- 11408485 TI - Differential molecular assemblies underlie the dual function of Axin in modulating the WNT and JNK pathways. AB - Axin is a multidomain scaffold protein that exerts a dual function in the Wnt signaling and MEKK1/JNK pathways. This raises a critical question as to whether Axin-based differential molecular assemblies exist and how these may act to coordinate the two separate pathways. Here we show that both wild-type glycogen synthase kinase-3 beta (GSK-3 beta) and kinase-dead GSK-3 beta-Y216F (capable of binding to Axin), but not GSK-3 beta-K85M (incapable of binding to Axin in mammalian cells), prevented MEKK1 binding to the Axin complex, thereby inhibiting JNK activation. We further show that casein kinase I epsilon also inhibited Axin mediated JNK activation by competing against MEKK1 binding. In contrast, beta catenin and adenomatous polyposis coli binding did not affect MEKK1 binding to the same Axin complex. This suggests that even when Axin is "switched" to activate the JNK pathway, it is still capable of sequestering free beta-catenin, which is a critical aspect for cellular homeostasis. Our results clearly demonstrate that differential molecular assemblies underlie the duality of Axin functions in the negative regulation of Wnt signaling and activation of the JNK MAPK pathway. PMID- 11408486 TI - Gln3p nuclear localization and interaction with Ure2p in Saccharomyces cerevisiae. AB - Gln3p is one of two well characterized GATA family transcriptional activation factors whose function is regulated by the nitrogen supply of the cell. When nitrogen is limiting, Gln3p and Gat1p are concentrated in the nucleus where they bind GATA sequences upstream of nitrogen catabolite repression (NCR)-sensitive genes and activate their transcription. Conversely, in excess nitrogen, these GATA sequences are unoccupied by Gln3p and Gat1p because these transcription activators are excluded from the nucleus. Ure2p binds to Gln3p and Gat1p and is required for NCR-sensitive transcription to be repressed and for nuclear exclusion of these transcription factors. Here we show the following. (i) Gln3p residues 344-365 are required for nuclear localization. (ii) Replacing Ser-344, Ser-347, and Ser-355 with alanines has minimal effects on GFP-Gln3p localization. However, replacing Gln3p Ser-344, Ser-347, and Ser-355 with aspartates results in significant loss of its ability to be concentrated in the nucleus. (iii) N and C termini of the Gln3p region required for it to complex with Ure2p and be excluded from the nucleus are between residues 1-103 and 301-365, respectively. (iv) N and C termini of the Ure2p region required for it to interact with Gln3p are situated between residues 101-151 and 330-346, respectively. (v) Loss of Ure2p residues participating in either dimer or prion formation diminishes its ability to carry out NCR-sensitive regulation of Gln3p activity. PMID- 11408487 TI - A lithium-induced conformational change in serotonin transporter alters cocaine binding, ion conductance, and reactivity of Cys-109. AB - Inactivation of serotonin transporter (SERT) expressed in HeLa cells by [2 (trimethylammonium)ethyl]methanethiosulfonate (MTSET) occurred much more readily when Na(+) in the reaction medium was replaced with Li(+). This did not result from a protective effect of Na(+) but rather from a Li(+)-specific increase in the reactivity of Cys-109 in the first external loop of the transporter. Li(+) alone of the alkali cations caused this increase in reactivity. Replacing Na(+) with N-methyl-d-glucamine (NMDG(+)) did not reduce the affinity of cocaine for SERT, as measured by displacement of a high affinity cocaine analog, but replacement of Na(+) with Li(+) led to a 2-fold increase in the K(D) for cocaine. The addition of either cocaine or serotonin (5-HT) protected SERT against MTSET inactivation. When SERT was expressed in Xenopus oocytes, inward currents were elicited by superfusing the cell with 5-HT (in the presence of Na(+)) or by replacing Na(+) with Li(+) but not NMDG(+). MTSET treatment of oocytes in Li(+) but not in Na(+) decreased both 5-HT and Li(+) induced currents, although 5-HT induced currents were inhibited to a greater extent. Na(+) antagonized the effects of Li(+) on both inactivation and current. These results are consistent with Li(+) inducing a conformational change that exposes Cys-109, decreases cocaine affinity, and increases the uncoupled inward current. PMID- 11408488 TI - TAT fusion proteins containing tyrosine 42-deleted IkappaBalpha arrest osteoclastogenesis. AB - In most circumstances, NF-kappaB, which is essential for osteoclastogenesis, is activated following serine 32/36 phosphorylation of its cytosolic inhibitory protein, IkappaBalpha. In contrast to other cell types, IkappaBalpha, in bone marrow macrophages (BMMs), which are osteoclast precursors, is tyrosine phosphorylated by c-Src kinase. To address the role of IkappaBalpha phosphorylation in osteoclastogenesis, we generated TAT fusion proteins containing wild-type IkappaBalpha (TAT-WT-IkappaB), IkappaBalpha lacking its NH(2)-terminal 45 amino acids (TAT-IkappaB(46-317)), and IkappaBalpha in which tyrosine residue 42, the c-Src target, is mutated into phenylalanine (TAT IkappaB(Y42F)). TAT-IkappaB efficiently enters BMMs, and the NF-kappaB-inhibitory protein, once intracellular, is functional. While TAT-WT-IkappaB only slightly inhibits osteoclastogenesis, osteoclast recruitment is diminished >80% by TAT IkappaB(46-317), an event mirrored by dentin resorption. The fact that TAT alone does not impact osteoclastogenesis, which also resumes following withdrawal of TAT-IkappaB(46-317), establishes that the mutant's anti-osteoclastogenic properties do not reflect toxicity. Affirming a functional role for IkappaB(Tyr(42)) in osteoclastogenesis, TAT-IkappaB(Y42F) is as efficient as TAT IkappaB(46-317) in blocking osteoclast differentiation. Thus, dominant-negative IkappaBalpha constructs block osteoclastogenesis, and Tyr(42) is essential to the process, increasing the possibility that nonphosphorylatable forms of IkappaBalpha may be a means of preventing pathological bone loss. PMID- 11408489 TI - Species-specific differences in amino acid editing by class II prolyl-tRNA synthetase. AB - Aminoacyl-tRNA synthetases are a family of enzymes responsible for ensuring the accuracy of the genetic code by specifically attaching a particular amino acid to their cognate tRNA substrates. Through primary sequence alignments, prolyl-tRNA synthetases (ProRSs) have been divided into two phylogenetically divergent groups. We have been interested in understanding whether the unusual evolutionary pattern of ProRSs corresponds to functional differences as well. Previously, we showed that some features of tRNA recognition and aminoacylation are indeed group specific. Here, we examine the species-specific differences in another enzymatic activity, namely amino acid editing. Proofreading or editing provides a mechanism by which incorrectly activated amino acids are hydrolyzed and thus prevented from misincorporation into proteins. "Prokaryotic-like" Escherichia coli ProRS has recently been shown to be capable of misactivating alanine and possesses both pretransfer and post-transfer hydrolytic editing activity against this noncognate amino acid. We now find that two ProRSs belonging to the "eukaryotic-like" group exhibit differences in their hydrolytic editing activity. Whereas ProRS from Methanococcus jannaschii is similar to E. coli in its ability to hydrolyze misactivated alanine via both pretransfer and post-transfer editing pathways, human ProRS lacks these activities. These results have implications for the selection or design of antibiotics that specifically target the editing active site of the prokaryotic-like group of ProRSs. PMID- 11408490 TI - Molecular basis of calmodulin tethering and Ca2+-dependent inactivation of L-type Ca2+ channels. AB - Ca(2+)-dependent inactivation (CDI) of L-type Ca(2+) channels plays a critical role in controlling Ca(2+) entry and downstream signal transduction in excitable cells. Ca(2+)-insensitive forms of calmodulin (CaM) act as dominant negatives to prevent CDI, suggesting that CaM acts as a resident Ca(2+) sensor. However, it is not known how the Ca(2+) sensor is constitutively tethered. We have found that the tethering of Ca(2+)-insensitive CaM was localized to the C-terminal tail of alpha(1C), close to the CDI effector motif, and that it depended on nanomolar Ca(2+) concentrations, likely attained in quiescent cells. Two stretches of amino acids were found to support the tethering and to contain putative CaM-binding sequences close to or overlapping residues previously shown to affect CDI and Ca(2+)-independent inactivation. Synthetic peptides containing these sequences displayed differences in CaM-binding properties, both in affinity and Ca(2+) dependence, leading us to propose a novel mechanism for CDI. In contrast to a traditional disinhibitory scenario, we suggest that apoCaM is tethered at two sites and signals actively to slow inactivation. When the C-terminal lobe of CaM binds to the nearby CaM effector sequence (IQ motif), the braking effect is relieved, and CDI is accelerated. PMID- 11408491 TI - Xyloside transport by XylP, a member of the galactoside-pentoside-hexuronide family. AB - This paper describes the functional characterization of the xyloside transporter, XylP, of Lactobacillus pentosus with the aid of a spectroscopy-based assay system. In order to monitor the transport reaction, the natural xyloside isoprimeverose, a building block of hemicellulose, and the analogue methyl isoprimeverose were chemically synthesized by a new and efficient procedure. The XylP protein was purified by metal affinity chromatography, following high level expression in Lactococcus lactis from the nisin-inducible promoter. The purified XylP protein was incorporated into liposomes, in which the glucose dehydrogenase from Acinetobacter calcoaceticus (sGDH) was entrapped. sGDH can oxidize aldose sugars in the presence of dichlorophenol-indophenol as electron acceptor. The coupled assay thus involves XylP-mediated isoprimeverose uptake followed by internal oxidation of the sugar by sGDH, which can be monitored from the reduction of 2,6-dichlorophenol-indophenol at 600 nm. The uptake of isoprimeverose was stimulated by the presence of the non-oxidizable methyl isoprimeverose on the trans-side of the membrane, indicating that exchange transport is faster than unidirectional downhill uptake. Unlike other members of the galactoside-pentoside-hexuronide family, XylP does not transport monosaccharides (xylose) but requires a glycosidic linkage at the anomeric carbon position. Consistent with a proton motive force-driven mechanism, the uptake was stimulated by a membrane potential (inside negative relative to outside) and inhibited by a pH gradient (inside acidic relative to outside). The advantages of the here-described transport assay for studies of carbohydrate transport are discussed. PMID- 11408492 TI - Phase II study of paclitaxel and valspodar (PSC 833) in refractory ovarian carcinoma: a gynecologic oncology group study. AB - PURPOSE: A phase II study was conducted to determine the efficacy of paclitaxel and valspodar (PSC 833) in patients with advanced epithelial ovarian cancer. Valspodar, a nonimmunosuppressive cyclosporine D analogue that reverses P glycoprotein-mediated multidrug resistance, in combination with paclitaxel might be active in paclitaxel-resistant and refractory ovarian cancer. PATIENTS AND METHODS: Patients received valspodar 5 mg/kg orally qid x 12 doses. Paclitaxel (70 mg/m(2) intravenously for 3 hours) was administered on day 2, 2 hours after the fifth or sixth dose of valspodar. This treatment was repeated every 21 days. One blood sample was collected before the sixth dose of valspodar for the first three cycles to evaluate valspodar trough concentration. Tumor tissue was obtained from patients for immunohistochemical staining of P-glycoprotein. RESULTS: Of 60 patients entered, 58 were assessable for response. There were five partial responses (8.6%; 90% confidence interval [CI], 3.8 to 20.0; median duration of response, 5.0 months [range, 1.9 to 10.5 months]). Median progression free survival was 1.5 months (90% CI, 1.4 to 2.4). Grade 3 or 4 toxicities observed were neutropenia, anemia, nausea and vomiting, peripheral neuropathy, and cerebellar ataxia. The trough concentrations of valspodar were > or = 1,000 ng/mL in all but two of 40 patients in the first cycle. Immunohistochemical staining for P-glycoprotein was positive for one of two responding patients. CONCLUSION: Valspodar in combination with paclitaxel has limited activity in patients with paclitaxel-resistant ovarian carcinoma. An international randomized clinical trial of paclitaxel and carboplatin with or without valspodar as first line therapy in advanced ovarian cancer is underway. PMID- 11408493 TI - Phase I/II trial of the multidrug-resistance modulator valspodar combined with cisplatin and doxorubicin in refractory ovarian cancer. AB - PURPOSE: To determine the maximum-tolerated dose (MTD) of doxorubicin when given in combination with cisplatin and the multidrug-resistance (MDR) modulator valspodar and the remission rate induced by this combination in patients with platinum- and anthracycline-resistant ovarian cancer. PATIENTS AND METHODS: Fifty nine patients who had failed prior platinum- and anthracycline-based chemotherapy were enrolled. During the dose-finding phase, patients received a loading dose of valspodar (1.5 or 2 mg/kg) via 2-hour intravenous (IV) infusion on day 1 and continuous IV infusion (CIVI) of valspodar (2, 4, or 10 mg/kg/d) over 3 days. Doxorubicin (starting from 20 up to 50 mg/m(2)) and cisplatin (50 mg/m(2)) were administered via 15- to 20-minute IV infusions on day 3. During the efficacy phase, patients received at least two treatment cycles unless toxicity was unacceptable, and responding patients and those with stable disease received four to six cycles. RESULTS: All patients completed at least one cycle of combined treatment. The MTD of doxorubicin was determined to be 35 mg/m(2) when administered with valspodar at 2 mg/kg loading dose and 10 mg/kg/d CIVI plus 50 mg/m(2) cisplatin. At these doses, valspodar blood concentrations known to reverse MDR in vitro were reached in all patients. Valspodar was well tolerated at all dose levels. Dose-limiting toxicities of the combination were primarily hematologic and included febrile neutropenia and prolonged leucopenia. The addition of valspodar to the treatment did not worsen cisplatin-related toxicity. Among 33 patients treated at the MTD for doxorubicin, one (3%) had a complete response, and four (12%) had a partial response. An additional seven patients experienced a stabilization of their previously progressive disease. The survival rates at 6 and 12 months were 59% and 19%, respectively. CONCLUSION: Valspodar can be safely coadministered with doxorubicin and cisplatin. Although the regimen used in this trial produced renewed responses in patients with heavily pretreated, refractory ovarian cancer, the value of valspodar in reversing resistance mediated by P-glycoprotein remains to be determined. PMID- 11408494 TI - Bone marrow features improve prognostic efficiency in multivariate risk classification of chronic-phase Ph(1+) chronic myelogenous leukemia: a multicenter trial. AB - PURPOSE: Multivariate risk classifications for chronic (stable)-phase Ph(1+) chronic myelogenous leukemia (CML) are generally focused on hematologic variables, and the putative prognostic property of bone morphology has been neglected or even contested so far. PATIENTS AND METHODS: A total of 510 consecutively recruited patients in first chronic phase Ph(1+) CML and pretreatment bone marrow biopsy specimens were entered onto this multicenter observational trial to evaluate the effect of bone marrow histopathology. According to generally accepted criteria, patients with any signs of accelerated disease were excluded. Treatment modalities included administration of interferon alfa-2b (IFN) and chemotherapy with hydroxyurea (HU) or busulfan. Immunohistochemical and morphometric techniques were applied to identify marrow cells and to quantify fiber density. Patients were separated into learning and validation samples, and classification and regression tree (CART) analysis was performed to establish a prognostic decision tree. RESULTS: CART analysis of the validation sample (123 patients with HU therapy) revealed the amount of erythroid precursors in the bone marrow, myelofibrosis, and splenomegaly as the most important prognostic features. Three risk profiles with significantly different survival patterns were established, with median survival times ranging from 33 to 108 months (two-sided log-rank test, P =.0001). The new score was confirmed by application to the learning sample with IFN therapy (two-sided log-rank test, P =.0002). Furthermore, risk status defined by the new score was significantly correlated with the occurrence of blast transformation. CONCLUSION: Our data strongly implicate that prognostic classification of chronic-phase Ph(1+) CML can be significantly improved by the inclusion of morphologic parameters. The variables of the presented scoring system may be easily assessed by routinely processed aspirates and bone marrow trephines. PMID- 11408495 TI - Combined interferon-alfa, 13-cis-retinoic acid, and alpha-tocopherol in locally advanced head and neck squamous cell carcinoma: novel bioadjuvant phase II trial. AB - PURPOSE: Retinoids and interferons (IFNs) have single-agent and synergistic combined effects in modulating cell proliferation, differentiation, and apoptosis in vitro and clinical activity in vivo in the head and neck and other sites. Alpha-tocopherol has chemopreventive activity in the head and neck and may decrease 13-cis-retinoic acid (13-cRA) toxicity. We designed the present phase II adjuvant trial to prevent recurrence or second primary tumors (SPTs) using 13 cRA, IFN-alpha, and alpha-tocopherol in locally advanced-stage head and neck cancer. PATIENTS AND METHODS: After definitive local treatment with surgery, radiotherapy, or both, patients with locally advanced SCCHN were treated with 13 cRA (50 mg/m(2)/d, orally, daily), IFN-alpha (3 x 10(6) IU/m(2), subcutaneous injection, three times a week), and alpha-tocopherol (1,200 IU/d, orally, daily) for 12 months, with a dose modification. Screening for recurrence or SPTs was performed every 3 months. RESULTS: Tumors of 11 (24%) of the 45 treated patients were stage III, and 34 (76%) were stage IV. Thirty-eight (86%) of 44 patients completed the full 12-month treatment (doses modified as needed). Toxicity generally was consistent with previous IFN and 13-cRA reports and included mild to moderate mucocutaneous and flu-like symptoms; occasional significant fatigue (grade 3 in 7% of patients), mild to moderate hypertriglyceridemia in 30% of patients who continued treatment along with antilipid therapy, and mild hematologic side effects. Six patients did not complete the planned treatment because of intolerable toxicity or social problems. At a median 24-months of follow-up, our clinical end point rates were 9% for local/regional recurrence (four patients), 5% for local/regional recurrence and distant metastases (two patients), and 2% for SPT (one patient), which was acute promyelocytic leukemia (ie, not of the upper aerodigestive tract). Median 1- and 2-year rates of overall survival were 98% and 91%, respectively, and of disease-free survival were 91% and 84%, respectively. CONCLUSION: The novel biologic agent combination of IFN alpha, 13-cRA, and alpha-tocopherol was generally well tolerated and promising as adjuvant therapy for locally advanced squamous cell carcinoma of the head and neck. We are currently conducting a phase III randomized study of this combination (v no treatment) to confirm these phase II study results. PMID- 11408496 TI - Paclitaxel and gemcitabine chemotherapy for advanced transitional-cell carcinoma of the urothelial tract: a phase II trial of the Minnie pearl cancer research network. AB - PURPOSE: To evaluate the toxicity and efficacy of combination chemotherapy with paclitaxel and gemcitabine in patients with advanced transitional-cell carcinoma of the urothelial tract. PATIENTS AND METHODS: Fifty-four patients with advanced unresectable urothelial carcinoma entered this multi-centered, community-based, phase II trial between May 1997 and December 1999. All patients were treated with paclitaxel 200 mg/m(2) by 1-hour intravenous (IV) infusion on day 1 and gemcitabine 1,000 mg/m(2) IV on days 1, 8, and 15; courses were repeated every 21 days. Patients who had objective response or stable disease continued treatment for six courses. RESULTS: Twenty-nine of 54 patients (54%; 95% confidence interval, 40% to 67%) had major responses to treatment, including 7% complete responses. With a median follow-up of 24 months, 16 patients (30%) remain alive and nine (17%) are progression-free. The median survival for the entire group was 14.4 months; 1- and 2-year actuarial survival rates were 57% and 25%, respectively. Seven (47%) of 15 patients previously treated with platinum-based chemotherapy responded to paclitaxel/gemcitabine. Grade 3/4 toxicity was primarily hematologic, including leukopenia (46%), thrombocytopenia (13%), and anemia (28%). Ten patients (19%) required hospitalization for neutropenia and fever, and one patient had treatment-related septic death. CONCLUSION: The combination of paclitaxel and gemcitabine is active and well tolerated in the first- or second-line treatment of patients with advanced transitional-cell carcinoma of the urothelial tract. Response rate and duration compare favorably with those produced by other active, first-line regimens. This regimen should be further evaluated in phase II and III studies, as well as in patients with compromised renal function. PMID- 11408497 TI - Prognostic significance of reverse transcriptase polymerase chain reaction for prostate-specific antigen in men with hormone-refractory prostate cancer. AB - PURPOSE: To evaluate the prognostic significance of reverse transcriptase polymerase chain reaction (RT-PCR) detection of prostate-specific antigen (PSA) mRNA in the blood of men with hormone-refractory prostate cancer (HRPC). PATIENTS AND METHODS: Peripheral blood was obtained from 193 men enrolled on Cancer and Leukemia Group B Study 9480, a prospective randomized comparison of three doses of suramin. RNA was isolated from the samples and assayed for the presence of PSA transcripts by RT-PCR. RESULTS: RNA could be isolated in 156 (83%) of samples. PSA transcripts as measured by RT-PCR were detectable in 75 (48%) of the 156 patients. The median survival for those patients in whom no transcripts were detectable was 18 months (95% confidence interval [CI], 14 to 22 months) compared with 13 months (95% CI, 11 to 15 months) (P =.004) for those in whom transcripts were detectable. In a multivariate analysis in which other factors predictive of survival were used, RT-PCR for PSA provided independent prognostic information. CONCLUSION: RT-PCR for PSA predicts survival duration in a population of men with HRPC. PMID- 11408498 TI - Clinical impact of germ cell tumor cells in apheresis products of patients receiving high-dose chemotherapy. AB - PURPOSE: High-dose chemotherapy (HD-Ctx) followed by autologous peripheral-blood stem-cell (PBSC) transplantation is currently investigated in patients with poor prognosis or relapsed metastatic germ cell tumor (GCTs). This study analyzed the presence and the clinical importance of contaminating tumor cells in PBSC preparations used to support HD-Ctx in GCT patients. PATIENTS AND METHODS: Seven targets for reverse transcription polymerase chain reaction (RT-PCR)-based detection of GCT cells were able to detect seminomatous and different histologic variants of nonseminomatous tumor cells. PBSC preparations from 57 patients were investigated for the presence of contaminating tumor cells using this set of targets, including beta human chorionic gonadotropin (beta-hCG), fibronectin (EDB variant), epidermal growth factor receptor (EGFR), CD44 (v8 to 10 variant), germ cell and placental alkaline phosphatase (AP), human endogenous retrovirus type K (ENV and GAG), and XIST. Samples of PBSC preparations from four healthy donors for allogenic transplantations as well as blood specimens from 10 healthy volunteers served as negative controls. RESULTS: Fifty patients (43 first-line and seven second-line Ctx) were assessable. Combining all RT-PCR results, 29 PBSC preparations (58%) were positive for tumor-specific amplification products (HERV K 0, fibronectin 4, XIST 14, beta-hCG 19, AP 19, CD44 24, EGFR 26). Ten (35%) of 29 patients who underwent transplantation with positive PBSC preparations and seven (33%) of 21 patients with negative PBSC preparations have suffered relapse or progression (not significant [ns]). With a median follow-up of 22 months (2 to 66) post-HD-Ctx projected 3-year survival rates are 68% (RT-PCR+) and 58% (RT-PCR ) (ns). None of the 10 control peripheral-blood samples showed positivity for any of the targets studied. CONCLUSION: GCT cells can be detected in more than 50% of PBSC preparations using a RT-PCR approach with multiple targets. Despite the presence of tumor cells, retransplantation of the PBSC products did not effect long-term outcome. Factors such as responsiveness to chemotherapy and tumor mass seem to overcome the importance of potentially re-infused tumor cells. PMID- 11408499 TI - Clinicopathologic features of long-term survivors and disease-free survivors after resection of hepatocellular carcinoma: a study of a prospective cohort. AB - PURPOSE: This study aims to clarify the clinicopathologic features of long-term survivors and disease-free survivors after resection of hepatocellular carcinoma (HCC). PATIENTS AND METHODS: The clinicopathologic features of 5-year survivors and disease-free survivors were elucidated in a cohort of 230 patients prospectively observed for > 5 years (64 to 192 months) after curative resection of HCC. RESULTS: The incidence of 5-year overall and disease-free survivors were 37% (85 of 230) and 20% (45 of 230), respectively. Clinicopathologic features associated with 5-year survivors included female sex (P =.024), preoperative serum albumin > or= 40 g/L (P =.033), AST < 50 u/L (P =.001), tumor < 5 cm (P =.001), solitary tumor (P =.035), encapsulated tumor (P =.021), no venous invasion (P =.001), no microsatellite nodule (P =.001), and early pathologic tumor-node-metastasis (pTNM) stage (I or II, P <.001). Features favoring 5-year disease-free survivors were preoperative serum AST < 50 u/L (P =.007), tumor < 5 cm (P =.005), encapsulated tumor (P =.007), no venous invasion (P <.001), no microsatellite nodule (P =.001), and early pTNM stage (I or II, P <.001). By multivariate analysis, pTNM stage was the only significant predictive factor for both overall and disease-free survival. CONCLUSION: This study shows that long term disease-free survival > 5 years after resection of HCC can be achieved in patients with favorable tumor characteristics. Early pTNM stage was the most reliable predictor of both long-term overall and disease-free survivors. PMID- 11408500 TI - Clinicopathologic re-evaluation of 100 malignant fibrous histiocytomas: prognostic relevance of subclassification. AB - PURPOSE: Malignant fibrous histiocytoma (MFH) has been regarded as the most common soft tissue sarcoma (STS) in adults. Yet its true nature and the validity of this diagnostic concept have increasingly been questioned. Available data suggest that most patients with MFH can be subclassified into specific STS types, but the clinical relevance of such categorization has been argued. In a retrospective study, we reclassified 100 tumors of the extremity and trunk wall primarily diagnosed as MFH and analyzed the outcome. PATIENTS AND METHODS: Patients were adults (median age, 70 years; range, 32 to 94 years). The median tumor size was 8 cm (range, 1 to 30 cm), and the thigh was the most common tumor location (n = 31). Median follow-up was 8 years (range, 3 to 16 years). The overall 5-year metastasis-free survival rate was 0.64. The tumors were reanalyzed histologically, immunohistochemically, and, where available, ultrastructurally, and were classified according to strict diagnostic criteria. Patients were staged according to the American Joint Committee on Cancer system, and prognoses were compared among different groups of the reclassified diagnoses, paying special attention to myogenic tumors. RESULTS: In 84 of 100 tumors, a specific line of differentiation was either proved or strongly suggested. The most common diagnoses were myxofibrosarcoma (n = 22) and leiomyosarcoma (n = 20). Overall, 30 tumors could be grouped as some form of myogenic sarcoma. These tumors had a worse prognosis, even within the same American Joint Committee on Cancer stage, and a shorter time to metastasis than nonmyogenic tumors. CONCLUSION: This retrospective study confirms that most so-called MFH can be subclassified by defined criteria; it provides evidence that such classification is clinically important. Specifically, pleomorphic STS showing myogenic differentiation are significantly more aggressive, a finding that allows planning future therapeutic trials. PMID- 11408501 TI - Prognostic value of initial fasting serum gastrin levels in patients with Zollinger-Ellison syndrome. AB - PURPOSE: To assess the value of the initial fasting serum gastrin (FSG) at presentation in patients with Zollinger-Ellison Syndrome (ZES) in predicting primary tumor characteristics and survival. PATIENTS AND METHODS: A total of 239 patients were treated for ZES between December 1981 and September 1998, with a mean follow-up of 9.1 +/- 0.6 years. At initial evaluation, 86 patients (36%) had mild (0 to 499 pg/mL), 61 (25.5%) had moderate (500 to 1,000 pg/mL), and 92 (38.5%) had severe (> 1,000 pg/mL) elevations in FSG. Primary tumor location and size, presence of lymph node or hepatic metastases, and survival were analyzed based on the level of initial FSG. RESULTS: In patients with sporadic ZES, but not in those with multiple endocrine neoplasia type 1 (MEN-1) and ZES, there was a significant relationship between the level of initial FSG and tumor size and location of primary tumor, frequency of lymph node and liver metastases, and survival. The median 5- and 10-year survival decreased with increasing initial FSG (P <.001) in patients with sporadic ZES; MEN-1 patients lived longer than sporadic ZES patients (P =.012), and survival in this group was not associated with the level of initial FSG. Multivariate analysis showed that factors independently associated with death from disease in patients with sporadic ZES were liver metastases (P =.0001), a pancreatic site (P =.0027), and primary tumor size (P =.011) but not initial FSG (P >.30). CONCLUSION: The severity of FSG at presentation is associated with size and site of tumor and the presence of hepatic metastases, factors that are significant independent predictors of outcome. The level of FSG at presentation may be useful in planning the nature and extent of the initial evaluation and management in patients with sporadic ZES. PMID- 11408502 TI - Prediction of response to preoperative chemotherapy in adenocarcinomas of the esophagogastric junction by metabolic imaging. AB - PURPOSE: Preoperative chemotherapy in patients with gastroesophageal cancer is hampered by the lack of reliable predictors of tumor response. This study evaluates whether positron emission tomography (PET) using fluorine-18 fluorodeoxyglucose (FDG) may predict response early in the course of therapy. PATIENTS AND METHODS: Forty consecutive patients with locally advanced adenocarcinomas of the esophagogastric junction were studied by FDG-PET at baseline and 14 days after initiation of cisplatin-based polychemotherapy. Clinical response (reduction of tumor length and wall thickness by > 50%) was evaluated after 3 months of therapy using endoscopy and standard imaging techniques. Patients with potentially resectable tumors underwent surgery, and tumor regression was assessed histopathologically. RESULTS: The reduction of tumor FDG uptake (mean +/- 1 SD) after 14 days of therapy was significantly different between responding (-54% +/- 17%) and nonresponding tumors (-15% +/- 21%). Optimal differentiation was achieved by a cutoff value of 35% reduction of initial FDG uptake. Applying this cutoff value as a criterion for a metabolic response predicted clinical response with a sensitivity and specificity of 93% (14 of 15 patients) and 95% (21 of 22), respectively. Histopathologically complete or subtotal tumor regression was achieved in 53% (eight of 15) of the patients with a metabolic response but only in 5% (one of 22) of the patients without a metabolic response. Patients without a metabolic response were also characterized by significantly shorter time to progression/recurrence (P =.01) and shorter overall survival (P =.04). CONCLUSION: PET imaging may differentiate responding and nonresponding tumors early in the course of therapy. By avoiding ineffective and potentially harmful treatment, this may markedly facilitate the use of preoperative therapy, especially in patients with potentially resectable tumors. PMID- 11408503 TI - Bony morbidity in children treated for acute lymphoblastic leukemia. AB - PURPOSE: Corticosteroids are widely used in the treatment of acute lymphoblastic leukemia (ALL). To determine the frequency of corticosteroid-associated bony morbidity in children with ALL, we retrospectively evaluated the incidence of fractures and osteonecrosis (ON) on two consecutive pediatric ALL protocols. PATIENTS AND METHODS: One hundred seventy-six consecutive children were treated for ALL between 1987 and 1995 at the Dana-Farber Cancer Institute and Children's Hospital. Prednisone was used as the corticosteroid during postremission therapy from 1987 to 1991, and dexamethasone was used from 1991 to 1995. Medical records for all patients were reviewed to assess the occurrence of fractures and ON. RESULTS: With a median follow-up of 7.6 years, the 5-year cumulative incidence (CI) +/- SE of any bony morbidity for the 176 patients was 30% +/- 4%, with a 5 year CI of fractures of 28% +/- 3% and of ON of 7% +/- 2%. With multivariate analysis, independent predictors of bony morbidity included age 9 to 18 years at diagnosis (P <.01), male sex (P <.01), and treatment with dexamethasone (P =.01). Dexamethasone was associated with a higher risk of fractures (5-year CI, 36% +/- 5% v 20% +/- 4% with prednisone; P =.04), but not ON (P =.40). The 5-year event free survival for the 176 patients was 79% +/- 3%. CONCLUSION: Children treated for ALL had a high incidence of fractures and ON. Older children, boys, and patients receiving dexamethasone were at increased risk for the development of bony morbidity. Future studies should attempt to minimize corticosteroid associated bony morbidity without compromising clinical efficacy. PMID- 11408504 TI - Improved outcome for patients with middle ear rhabdomyosarcoma: a children's oncology group study. AB - PURPOSE: The goal of this study was to define the clinical features and optimal therapy for children and adolescents with middle ear (ME) rhabdomyosarcoma (RMS). PATIENTS AND METHODS: We reviewed demographic data, clinical features, therapy (including chemotherapy, surgery, and radiation), and outcome for the 179 eligible patients with ME RMS who were enrolled onto Intergroup Rhabdomyosarcoma Studies (IRS) I through IV or pilot studies between November 1972 and December 1997. RESULTS: Most patients were younger than 10 years old (90%), and 63% were male. Because of the parameningeal location, most tumors were not resected before chemotherapy (group I, < 1%; group II, 4%; group III, 84%; group IV, 12%). Although most tumors were locally invasive (T2, 89%), the majority were small (< or = 5 cm, 66%), lacked nodal metastases (N0, 86%), and had embryonal histology (85%). The 5-year failure-free survival (FFS) and overall survival (OS) estimates were 67% and 72%, respectively. Both FFS and OS improved significantly over the course of IRS I through IV (3-year FFS and OS: IRS-I, 42% and 42%; IRS-II, 70% and 74%; IRS-III, 65% and 72%; IRS-IV pilot, 81% and 96%; IRS-IV, 88% and 88%, P <.001). Lower clinical group or stage and smaller tumor size were associated with better outcome. Age, sex, tumor invasiveness, and nodal metastases were not predictive of outcome. CONCLUSION: Patients with ME RMS generally present with small, unresectable, invasive tumors at a site traditionally considered prognostically unfavorable. Nevertheless, such patients have benefited markedly from improvements in multimodal, risk-based therapy during the course of IRS I through IV, and with contemporary therapy, most are cured. PMID- 11408505 TI - Comprehensive genetic and histopathologic study reveals three types of neuroblastoma tumors. AB - PURPOSE: To determine the relationship between multiple genetic features, tumor morphology, and prognosis in neuroblastoma. PATIENTS AND METHODS: The genetic alterations and morphologic features that underpin three histopathologic risk classifications were analyzed in 108 neuroblastoma patients. Tumors were subdivided into four groups based on the three most frequent and prognostically significant genetic alterations (17q gain, 1p deletion, and MYCN amplification), and all other genetic, morphologic, and clinical data were analyzed with respect to these groups. RESULTS: Our analyses identify three nonoverlapping tumor types with distinct genetic and morphologic features, defined here as types 1, 2, and 3. Type 1 tumors show none of the three significant genetic alterations and have good prognosis. Both type 2 (17q gain only or 17q gain and 1p del) and type 3 (17q gain, 1p del, and MYCN amplification) tumors progress. However, these tumor types are distinguished clinically by having significantly different median age at diagnosis and median progression-free survival (PFS). Multivariate analysis indicates that 17q gain is the only independent prognostic factor among all genetic, histopathologic, and clinical factors analyzed. Among histopathologic risk systems, the International Neuroblastoma Pathology Classification was the best predictor of PFS. CONCLUSION: Our results indicate that specific combinations of genetic changes in neuroblastoma tumors contribute to distinct morphologic and clinical features. Furthermore, the identification of two genetically and morphologically distinct types of progressing tumors suggests that possibilities for different therapeutic regimens should be investigated. PMID- 11408506 TI - Intergroup rhabdomyosarcoma study-IV: results for patients with nonmetastatic disease. AB - PURPOSE: The study goal was to improve outcome in children with rhabdomyosarcoma by comparing risk-based regimens of surgery, radiotherapy (RT) and chemotherapy. PATIENTS AND METHODS: Eight hundred eighty-three previously untreated eligible patients with nonmetastatic rhabdomyosarcoma entered the Intergroup Rhabdomyosarcoma Study-IV (IRS-IV) (1991 to 1997) after surgery and were randomized treatment by primary tumor site, group (1 to 3), and stage (I to III). Failure-free survival (FFS) rates and survival were the end points used in comparisons between randomized groups and between patient subgroups treated on IRS-III and IRS-IV. Most patients were randomized to receive vincristine and dactinomycin (VA) and cyclophosphamide (VAC, n = 235), or VA and ifosfamide (VAI, n = 222), or vincristine, ifosfamide, and etoposide (VIE, n = 236). Patients with group 3 tumors were randomized to receive conventional RT (C-RT) versus hyperfractionated RT (HF-RT). RESULTS: Overall 3-year FFS and survival were 77% and 86%, respectively. Three-year FFS rates with VAC, VAI, and VIE were 75%, 77%, and 77%, respectively (P =.42). No significant difference in outcome was noted with HF-RT versus C-RT (P =.85 and P =.90, respectively). Overall, patients with embryonal tumors benefited from intensive three-drug chemotherapy in IRS-IV (3 year FFS, 83%). The improvement was seen for patients with stage I or stage II/III, group 1/2 disease, many of whom received VA chemotherapy on IRS-III. Patients with stage 2/3, group 3 disease had similar outcomes on IRS-III and IRS IV. Three-year FFS for the nonrandomized patient subsets was 75% with renal abnormalities; 81% for paratesticular, group 1 cases; and 91% for group 1/2 orbit or eyelid tumors. Patients with paratesticular primaries had poorer outcomes if they were more than 10 years old (3-year FFS, 63% v 90%). Myelosuppression occurred in most patients, but toxic deaths occurred in less than 1%. CONCLUSION: VAC and VAI or VIE with surgery (with or without RT), are equally effective for patients with local or regional rhabdomyosarcoma and are more effective for embryonal tumors than therapies used previously. Younger patients with group 1 paratesticular embryonal tumors and all patients with group 1/2 orbit or eyelid tumors can usually be cured with VA chemotherapy along with postoperative RT for group 2 disease. PMID- 11408507 TI - Phase III trial comparing two dose levels of epirubicin combined with cyclophosphamide with cyclophosphamide, methotrexate, and fluorouracil in node positive breast cancer. AB - PURPOSE: To compare a full-dose epirubicin-cyclophosphamide (HEC) regimen with classical cyclophosphamide, methotrexate, and fluorouracil (CMF) therapy and with a moderate-dose epirubicin-cyclophosphamide regimen (EC) in the adjuvant therapy of node-positive breast cancer. PATIENTS AND METHODS: Node-positive breast cancer patients who were aged 70 years or younger were randomly allocated to one of the following treatments: CMF for six cycles (oral cyclophosphamide); EC for eight cycles (epirubicin 60 mg/m(2), cyclophosphamide 500 mg/m(2); day 1 every 3 weeks); and HEC for eight cycles (epirubicin 100 mg/m(2), cyclophosphamide 830 mg/m(2); day 1 every 3 weeks). RESULTS: Two hundred fifty-five, 267, and 255 eligible patients were treated with CMF, EC, and HEC, respectively. Patient characteristics were well balanced among the three arms. One and three cases of congestive heart failure were reported in the EC and HEC arms, respectively. Three cases of acute myeloid leukemia were reported in the HEC arm. After 4 years of median follow-up, no statistically significant differences were observed between HEC and CMF (event-free survival [EFS]: hazards ratio [HR] = 0.96, 95% confidence interval [CI], 0.70 to 1.31, P =.80; distant-EFS: HR = 0.97, 95% CI, 0.70 to 1.34, P =.87; overall survival [OS]: HR = 0.97, 95% CI, 0.65 to 1.44, P =.87). HEC is more effective than EC (EFS: HR = 0.73, 95% CI, 0.54 to 0.99, P =.04; distant-EFS: HR = 0.75, 95% CI, 0.55 to 1.02, P =.06; OS HR = 0.69, 95% CI, 0.47 to 1.00, P =.05). CONCLUSION: This three-arm study does not show an advantage in favor of an adequately dosed epirubicin-based regimen over classical CMF in the adjuvant therapy of node-positive pre- and postmenopausal women with breast cancer. Moreover, this study confirms that there is a dose-response curve for epirubicin in breast cancer adjuvant therapy. PMID- 11408508 TI - Indicators of lifetime estrogen exposure: effect on breast cancer incidence and interaction with raloxifene therapy in the multiple outcomes of raloxifene evaluation study participants. AB - PURPOSE: To test the hypothesis that risk factors related to lifetime estrogen exposure predict breast cancer incidence and to test if any subgroups experience enhanced benefit from raloxifene. PATIENTS AND METHODS: Postmenopausal women with osteoporosis (N = 7,705), enrolled onto the Multiple Outcomes of Raloxifene Evaluation (MORE) trial, were randomly assigned to receive placebo, raloxifene 60 mg/d, or raloxifene 120 mg/d for 4 years. Breast cancer risk was analyzed by the following baseline characteristics indicative of estrogen exposure: previous hormone replacement therapy, prevalent vertebral fractures, family history of breast cancer, estradiol level, bone mineral density (BMD), body mass index, and age at menopause. Therapy-by-subgroup interactions were assessed using a logistic regression model. RESULTS: Overall, women with the highest one-third estradiol levels (> or = 12 pmol/L) had a 2.07-fold increased invasive breast cancer risk compared with women with lower levels. Raloxifene significantly reduced breast cancer risk in both the low- and high-estrogen subgroups for all risk factors examined (P <.05 for each comparison). The women with the highest BMD and those with a family history of breast cancer experienced a significantly greater therapy benefit with raloxifene, compared with the two thirds of patients with lower BMD or those without a family history, respectively; the subgroup-by therapy interactions were significant (P =.005 and P =.015, respectively). CONCLUSION: The MORE trial confirms that increased lifetime estrogen exposure increases breast cancer risk. Raloxifene therapy reduces breast cancer risk in postmenopausal osteoporotic women regardless of lifetime estrogen exposure, but the reduction is greater in those with higher lifetime exposure to estrogen. PMID- 11408509 TI - Phase I trial of pegylated liposomal doxorubicin and docetaxel in advanced breast cancer. AB - PURPOSE: To develop a combination of pegylated liposomal doxorubicin (Doxil; Alza Pharmaceuticals, Palo Alto, CA) and docetaxel (Taxotere; Aventis Pharmaceutical, Parsipanny, NJ) that can be safely used for the treatment of advanced breast cancer. PATIENTS AND METHODS: Forty-one patients with locally advanced (n = 10) or metastatic (n = 31) breast cancer received Doxil (30-, 40-, or 45-mg/m(2) intravenous [IV] infusion over 30 to 60 minutes), followed 1 hour later by docetaxel (60 or 75 mg/m(2) by IV infusion over 1 hour) in cohorts of three to six patients. Dose-limiting toxicity (DLT) was defined as febrile neutropenia, prolonged neutropenia, or grade 3 to 4 nonhematologic toxicity that occurred during cycle 1. RESULTS: In conjunction with docetaxel 75 mg/m(2) every 4 weeks, the MTD of Doxil was 30 mg/m(2) and required granulocyte colony-stimulating factor (G-CSF) to prevent febrile neutropenia. Without G-CSF, the MTD was docetaxel 60 mg/m(2) and Doxil 30 mg/m(2) every 3 weeks; only 1 (7%) out of 15 patients treated at this dose level had cycle 1 DLT. Infusion reactions were common with Doxil with the recommended infusion schedule during the first cycle (55%) but were reduced with a modified schedule (7%). There was no clinically significant cardiac toxicity. Objective response occurred in eight of nine assessable patients with stage III disease and in 16 (52%) of 31 patients (95% confidence interval, 34% to 70%) with stage IV disease. CONCLUSION: The recommended dose and schedule of this combination for further evaluation is Doxil 30 mg/m(2) and docetaxel 60 mg/m(2) given every 3 weeks without G-CSF. When used with G-CSF, it is Doxil 30 mg/m(2) and docetaxel 75 mg/m(2) every 4 weeks. PMID- 11408510 TI - Carboplatin skin testing: a skin-testing protocol for predicting hypersensitivity to carboplatin chemotherapy. AB - PURPOSE: A high incidence of moderate to severe hypersensitivity reactions (HRs) is noted in patients who have been treated with multiple courses of carboplatin. Presently, there is no reliable way to predict which patients may be at risk for this potentially severe adverse reaction. We developed a skin-test protocol to identify patients at high risk for HR to carboplatin chemotherapy. PATIENTS AND METHODS: Patients undergoing more than seven courses of carboplatin received a 0.02-mL intradermal injection of an undiluted aliquot of their planned carboplatin infusion 1 hour before each course of the agent. A positive skin test was prospectively defined as that resulting in a wheel of at least 5 mm with a surrounding flare. We recently reported a 27% incidence of HRs in patients receiving more than seven courses of carboplatin. These patients served as historical controls for the current study. RESULTS: Forty-seven patients with recurrent ovarian or primary peritoneal carcinoma receiving carboplatin were skin tested. Thirteen of 47 patients (28%) manifested a positive skin test at a median of nine total courses of carboplatin (range, eight to 17 courses). This rate of skin-test positivity was not significantly different from the incidence of documented HR reported in a historical control group (P =.89), suggesting comparable populations. A negative skin test accurately predicted the absence of HR in 166 of 168 courses of chemotherapy. Only two of 47 patients (4%) experienced a HR after a negative skin test. Thus, administering carboplatin only to patients with a negative skin test may result in a significant reduction in HRs relative to historical controls (P =.002). CONCLUSION: An easily performed skin test appears to predict patients in whom carboplatin may be safely administered. Treatment modifications based on the results of skin testing may reduce the incidence of HRs in patients receiving repeated courses of carboplatin. PMID- 11408511 TI - Phase I and pharmacokinetic study of the novel MDR1 and MRP1 inhibitor biricodar administered alone and in combination with doxorubicin. AB - PURPOSE: To evaluate the safety, tolerability, and pharmacokinetics of biricodar (VX-710), an inhibitor of P-glycoprotein (P-gp) and multidrug resistance associated protein (MRP1), alone and with doxorubicin in patients with advanced malignancies. The effect of VX-710 on the tissue distribution of (99m)Tc sestamibi, a P-gp and MRP1 substrate, was also evaluated. PATIENTS AND METHODS: Patients with solid malignancies refractory to standard therapy first received a 96-hour infusion of VX-710 alone at 20 to 160 mg/m(2)/h. After a 3-day washout, a second infusion of VX-710 was begun, on the second day of which doxorubicin 45 mg/m(2) was administered. Cycles were repeated every 21 to 28 days. (99m)Tc sestamibi scans were performed before and during administration of VX-710 alone. RESULTS: Of the 28 patients who enrolled, 25 patients were eligible for analysis. No dose-limiting toxicity (DLT) was observed in the nine assessable patients who received 120 mg/m(2)/h or less. Among seven patients receiving VX-710 160 mg/m(2)/h, two DLTs were seen: reversible CNS toxicity and febrile neutropenia. All other adverse events were mild to moderate and reversible. Plasma concentrations of VX-710 in patients who received at 120 and 160 mg/m(2)/h were two- to fourfold higher than concentrations required to fully reverse drug resistance in vitro. VX-710 exhibited linear pharmacokinetics with a harmonic mean half-life of 1.1 hours. VX-710 enhanced hepatic uptake and retention of (99m)Tc-sestamibi in all patients. CONCLUSION: A 96-hour infusion of VX-710 at 120 mg/m(2)/h plus doxorubicin 45 mg/m(2) has acceptable toxicity in patients with refractory malignancies. The safety and pharmacokinetics of VX-710 plus doxorubicin warrant efficacy trials in malignancies expressing P-gp and/or MRP1. PMID- 11408512 TI - The case for a new national program for the development of cancer therapeutics. PMID- 11408513 TI - Clinical trial designs for cytostatic agents. PMID- 11408514 TI - Does detection of circulating ONYX-015 genome by polymerase chain reaction indicate vector replication? PMID- 11408515 TI - Assessing adjuvant breast cancer therapy benefit. PMID- 11408516 TI - Measurement of brain tumor volumes by the perimeter method. PMID- 11408517 TI - Prostate-specific antigen levels in nipple aspirate fluid. PMID- 11408518 TI - The dopamine transporter and cocaine medication development: drug self administration in nonhuman primates. AB - Despite intensive medication development efforts, no effective pharmacotherapy for cocaine abuse has demonstrated efficacy for long-term use. Given the obvious importance of the dopamine transporter in the addictive properties of cocaine, the development and use of compounds that target the dopamine transporter represents a reasonable approach for the pharmacological treatment of cocaine abuse. The therapeutic approach of replacement or substitute agonist medication has been successful, as shown with methadone maintenance for heroin dependence and nicotine replacement for tobacco use. A number of preclinical studies with dopamine transporter inhibitors provide evidence that substitute agonists may be used effectively to reduce cocaine use. Nonhuman primate models of drug self administration provide a rigorous, systematic approach to characterize medication effectiveness in subjects with a documented history of drug use. Several cocaine analogs and other dopamine transporter inhibitors, including analogs of GBR 12909 and WIN 35,065-2, have been shown to reduce cocaine self-administration in nonhuman primates. A possible limitation to the use of selective dopamine transporter inhibitors as medications is their potential for abuse liability given their demonstrated reinforcing effects in nonhuman primates. However, limited reinforcing properties in the context of treatment programs may be advantageous, contributing to improved patient compliance and enhanced medication effectiveness. Moreover, pharmacokinetic properties that result in slow onset and long duration of action may enhance their effectiveness to reduce cocaine use while limiting their abuse liability. PMID- 11408519 TI - Mechanisms of endocannabinoid inactivation: biochemistry and pharmacology. AB - The endocannabinoids, a family of endogenous lipids that activate cannabinoid receptors, are released from cells in a stimulus-dependent manner by cleavage of membrane lipid precursors. After release, the endocannabinoids are rapidly deactivated by uptake into cells and enzymatic hydrolysis. Endocannabinoid reuptake occurs via a carrier-mediated mechanism, which has not yet been molecularly characterized. Endocannabinoid reuptake has been demonstrated in discrete brain regions and in various tissues and cells throughout the body. Inhibitors of endocannabinoid reuptake include N-(4-hydroxyphenyl) arachidonylamide (AM404), which blocks transport with IC50 (concentration necessary to produce half-maximal inhibition) values in the low micromolar range. AM404 does not directly activate cannabinoid receptors or display cannabimimetic activity in vivo. Nevertheless, AM404 increases circulating anandamide levels and inhibits motor activity, an effect that is prevented by the CB1 cannabinoid antagonist N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl 1H-pyrazole-3-carboxamide hydrochloride (SR141716A). AM404 also reduces behavioral responses to dopamine agonists and normalizes motor activity in a rat model of attention deficit hyperactivity disorder. The endocannabinoids are hydrolyzed by an intracellular membrane-bound enzyme, termed anandamide amidohydrolase (AAH), which has been molecularly cloned. Several fatty acid sulfonyl fluorides inhibit AAH activity irreversibly with IC50 values in the low nanomolar range and protect anandamide from deactivation in vivo. alpha-Keto oxazolopyridines inhibit AAH activity with high potency (IC50 values in the low picomolar range). A more thorough characterization of the roles of endocannabinoids in health and disease will be necessary to define the significance of endocannabinoid inactivation mechanisms as targets for therapeutic drugs. PMID- 11408520 TI - The anticonvulsant, antihyperalgesic agent gabapentin is an agonist at brain gamma-aminobutyric acid type B receptors negatively coupled to voltage-dependent calcium channels. AB - Gabapentin (Neurontin, Pfizer Global R & D) is a novel anticonvulsant, antihyperalgesic, and antinociceptive agent with a poorly understood mechanism of action. In this study, we show that gabapentin (EC50 2 microM) inhibited up to 70 to 80% of the total K+-evoked Ca2+ influx via voltage-dependent calcium channels (VD-CCs) in a mouse pituitary intermediate melanotrope clonal mIL-tsA58 (mIL) cell line. mIL cells endogenously express only gamma-aminobutyric acid type B (GABA(B)) gb1a-gb2 receptors. Moreover, activity of the agonist gabapentin was dose dependently and completely blocked with the GABA(B) antagonist CGP55845 and was nearly identical to the prototypic GABA(B) agonist baclofen in both extent and potency. Antisense knockdown of gb1a also completely blocked gabapentin activity, while gb1b antisense and control oligonucleotides had no effect, indicating that gabapentin inhibition of membrane Ca2+ mobilization in mIL cells was dependent on a functional GABA(B) (gb1a-gb2) heterodimer receptor. In addition, during combined whole cell recording and multiphoton Ca2+ imaging in hippocampal neurons in situ, gabapentin significantly inhibited in a dose dependent manner subthreshold soma depolarizations and Ca2+ responses evoked by somatic current injection. Furthermore, gabapentin almost completely blocked Ca2+ action potentials and Ca2+ responses elicited by suprathreshold current injection. However, larger current injection overcame this inhibition of Ca2+ action potentials suggesting that gabapentin did not predominantly affect L-type Ca2+ channels. The depressant effect of gabapentin on Ca2+ responses was coupled to the activation of neuronal GABA(B) receptors since they were blocked by CGP55845, and baclofen produced similar effects. Thus gabapentin activation of neuronal GABA(B) gb1a-gb2 receptors negatively coupled to VD-CCs can be a potentially important therapeutic mechanism of action of gabapentin that may be linked to inhibition of neurotransmitter release in some systems. PMID- 11408521 TI - In vivo inhibition of Fas ligand-mediated killing by TR6, a Fas ligand decoy receptor. AB - TR6, a member of the tumor necrosis factor (TNF) receptor superfamily, has recently been shown to bind to Fas ligand (FasL) and inhibit FasL-mediated cell killing in vitro. In the current study, we demonstrate that TR6 can block the lethal activity of FasL in multiple in vitro systems, and extend this finding to an in vivo model of hepatitis. The binding of human TR6 to human FasL was verified with BIAcore chip technology. Human primary hepatocytes, HT-29 cells and Jurkat cells were assayed for viability to demonstrate TR6 inhibition of FasL mediated cytotoxicity in vitro. Human TR6 was also shown to cross-react with membrane-bound mouse FasL, since the in vitro cytotoxic activity of L929 cells transfected with murine FasL was inhibited in the presence of human TR6. In vivo, FasL-induced acute, lethal, fulminant hepatic apoptosis resulting in death within 2 h of intravenous injection into Fas+ mice, but not Fas- MRL/lpr mice. Pretreatment of mice with TR6 blocked FasL-induced mortality, presumably by attenuating FasL-induced hepatic apoptosis. Thus, in both in vitro and in vivo systems, TR6 acts as a functional FasL decoy receptor and may be clinically useful in the treatment of hepatitis and other diseases associated with FasL mediated tissue injury. PMID- 11408522 TI - Administration of a potent antagonist of protease-activated receptor-1 (PAR-1) attenuates vascular restenosis following balloon angioplasty in rats. AB - Human platelets possess two distinct thrombin-activated receptors, PAR-1 (protease-activated receptor-1) and PAR-4, whereas human vascular smooth muscle cells possess only PAR-1. Although such thrombin receptors have been studied extensively in vitro, their physiological roles are still rather ill-defined. We have now employed a potent, selective PAR-1 antagonist, RWJ-58259, to probe the in vivo significance of PAR-1 in thrombosis and vascular injury. RWJ-58259 was examined in two thrombosis models in guinea pigs: the arteriovenous (A-V) shunt assay (monitoring thrombus weight) and the Rose Bengal intravascular photoactivation assay (monitoring time to occlusion). Administration of RWJ-58259 (10 mg/kg, total i.v. dose) did not inhibit thrombus formation in either thrombosis model, although local, intrashunt delivery in the A-V shunt model did elicit a modest antithrombotic effect (thrombus weight reduction from 35 +/- 2 to 24 +/- 4 mg). These results are consistent with the presence of more than one thrombin-sensitive receptor on guinea pig platelets, in analogy with human platelets. Indeed, we were able to establish that guinea pig platelets express three thrombin receptors, PAR-1, PAR-3, and PAR-4. We also examined RWJ-58259 in a vascular restenosis model involving balloon angioplasty in rats. Perivascular administration of RWJ-58259 (10 mg) significantly reduced neointimal thickness (77 +/- 5 microm to 45 +/- 5 microm, P < 0.05), clearly demonstrating an important role for PAR-1 in vascular injury. From these results, it is evident that a PAR-1 antagonist is not especially effective for treating platelet dependent thrombosis; however, it could well be beneficial for treating restenosis attendant to arterial injury. PMID- 11408523 TI - Multiple pathways of angiotensin I conversion and their functional role in the canine penile corpus cavernosum. AB - Multiple pathways of angiotensin (Ang) I conversion and their functional role in the canine penile corpus cavernosum were investigated. Biochemical analysis revealed high activities of angiotensin-converting enzyme (ACE) (6.9 +/- 1.7 mU/mg of protein, mean +/- S.E.M., n = 8) and chymase-like enzyme (4.0 +/- 1.4 mU/mg of protein). Functional recording of isometric tension showed that Ang I (3 x 10(-7) M) induced a tension of 0.17 +/- 0.05 g (n = 5), which was reduced to about 60% by pretreatment with an ACE inhibitor, lisinopril (10(-6) M), and almost completely blocked by lisinopril in combination with a chymase inhibitor, chymostatin (10(-4) M). Binding sites for ACE and Ang II receptors were studied by in vitro autoradiography using 125I-351A and 125I-[Sar1, Ile8]Ang II as ligands, respectively. Dense binding of ACE appeared in the endothelial layer of the corpus cavernosum penis, and Ang II receptors were localized in the trabecular smooth muscle layer. An AT1 receptor antagonist, CV-11974 (10(-6) M), markedly displaced 125I-[Sar1, Ile8]Ang II bindings, indicating that the corpus cavernosum penis contains AT1 receptors exclusively. Immunohistochemical studies demonstrated ACE in the endothelium of the corpus cavernosum penis. Mast cells that produce chymase were present mainly in the cavernosal area. These results demonstrate that chymase, in addition to ACE, is involved in the contraction of canine penile corpus cavernosum through local Ang II formation. PMID- 11408524 TI - Molecular basis of perinatal changes in UDP-glucuronosyltransferase activity in maternal rat liver. AB - The molecular basis of perinatal changes occurring in major UDP glucuronosyltransferase (UGT) family 1 isoforms and in UGT2B1, a relevant isoform belonging to family 2, was analyzed in rat liver. Nonpregnant, pregnant (19-20 days of pregnancy), and two groups of postpartum animals corresponding to early and middle stages of lactation (2-4 and 10-12 days after delivery, respectively) were studied. UGT activity determined in UDP-N-acetylglucosamine-activated microsomes revealed that bilirubin, p-nitrophenol, and ethynylestradiol (17beta OH and 3-OH) but not androsterone and estrone glucuronidation rates, were decreased in pregnant rats. Decreased enzyme activities returned to control values after delivery. p-Nitrophenol, androsterone, and estrone conjugation rate increased in postpartum rats. Western blot analysis performed with anti-peptide specific (anti-1A1, 1A5, 1A6, and 2B1) antibodies revealed decreased levels of all family 1 isoforms and UGT2B1 during pregnancy. In postpartum animals, protein level recovered (1A5 and 2B1) or even increased (1A1 and 1A6) with respect to control rats. Northern blot analysis suggested that expression of UGT proteins is down-regulated at a post-translational level during pregnancy and that increased levels of 1A1 and 1A6 observed in postpartum rats were associated to increased mRNA. To establish whether prolactin is involved in up-regulation of UGT1A1 and 1A6 postpartum, ovariectomized rats were treated with 300 microg of ovine prolactin per day for 7 days. The data indicated that prolactin was able to increase expression of UGT1A6 (protein and mRNA) but not 1A1. Thus, prolactin is the likely mediator of the increased expression of UGT1A6 observed in maternal liver postpartum. PMID- 11408525 TI - In vivo pharmacokinetics of selective mu-opioid peptide agonists. AB - Recent evidence suggests that highly selective mu-opioid agonists may provide good analgesia with less development of tolerance and dependence. H-Tyr-D-Arg-Phe Lys-NH2 (DALDA) and H-Dmt-D-Arg-Phe-Lys-NH2 ([Dmt1]DALDA) were found to display high binding affinity and much greater selectivity for the mu-opioid receptor (K(i)delta/K(i)mu) > 10,000) compared with H-Tyr-D-Ala-Gly-MePhe-Gly-ol (DAMGO). In addition, [Dmt1]DALDA was 3000-fold more potent than morphine when administered intrathecally. A potential problem with peptide analogs as therapeutic agents is their susceptibility to enzymatic degradation in vivo and short elimination half-lives. In this study, we compared the stability of DAMGO, DALDA, and [Dmt1]DALDA after systemic administration in sheep. Peptide concentrations were measured using high performance liquid chromatography-mass spectrometry. When incubated in sheep blood at 37 degrees C, DAMGO, DALDA, and [Dmt1]DALDA were stable over 2 h. When given intravenously to sheep, the apparent volume of distribution was 50 to 80 ml/kg for all three peptides, suggesting that distribution was limited to blood volume. Plasma clearance of DAMGO (223 ml/kg/h) was 10-fold faster than DALDA and [Dmt1]DALDA (24 ml/kg/h), and their elimination half-lives were 0.24, 1.5, and 1.8 h, respectively. The half-lives of DALDA and [Dmt1]DALDA are even longer than morphine or meperidine in sheep. These favorable pharmacokinetic properties of DALDA and [Dmt1]DALDA, together with their mu selectivity, potency, and long duration of action, make them ideal candidates as opioid analgesics. PMID- 11408526 TI - AT1 receptor antagonist telmisartan administered peripherally inhibits central responses to angiotensin II in conscious rats. AB - The effects of systemic treatment with the AT1 receptor antagonist telmisartan on central effects of angiotensin II (Ang II), namely, increase in blood pressure, vasopressin release into the circulation, and drinking response, were investigated in conscious, normotensive rats. The central responses to i.c.v. Ang II (30 ng/kg) were measured at 0.5, 2, 4, and 24 h following acute i.v. or acute and chronic oral telmisartan application. At a dose of 10 mg/kg i.v., the drinking response to i.c.v. Ang II was completely blocked over 4 h, while the pressor response and the release of vasopressin in response to i.c.v. Ang II were blocked by 60 to 80%. The inhibition of the centrally mediated pressor and drinking response to Ang II was sustained over 24 h. The lower doses of telmisartan (0.3 and 1 mg/kg) significantly inhibited the Ang II-induced actions over 4 h. A consistent 24-h inhibition of the central responses to i.c.v. Ang II was obtained after acute and chronic oral treatment with 30 mg/kg telmisartan. Oral treatment with 1 and 3 mg/kg telmisartan produced a slight but inconsistent inhibition of the central actions of Ang II. Telmisartan concentrations measured in the cerebrospinal fluid following 8 days of consecutive daily oral treatment (1-30 mg/kg) ranged from 0.87 +/- 0.27 ng/ml (1 mg/kg/day) to 46.5 +/- 11.6 ng/ml (30 mg/kg/day). Our results demonstrate that, following peripheral administration, the AT1 receptor antagonist telmisartan can penetrate the blood brain barrier in a dose- and time-dependent manner to inhibit centrally mediated effects of Ang II. PMID- 11408527 TI - Histamine increases interstitial adenosine concentration via activation of ecto 5'-nucleotidase in rat hearts in vivo. AB - We examined whether histamine enhances the production of interstitial adenosine via stimulation of ecto-5'-nucleotidase (a key enzyme responsible for adenosine production) using microdialysis techniques in in situ rat hearts. The microdialysis probe was implanted in the left ventricular myocardium of anesthetized rats and perfused in the presence of adenosine 5'-monophosphate (AMP). Histamine (10-500 microM) administered into the perfusate had a tendency to increase the adenosine concentration. In the presence of prazosin (50 microM), an antagonist of alpha1-adrenoceptors, or of chelerythrine (10 microM), a protein kinase C (PKC) inhibitor, and in reserpinized rats, histamine failed to increase the AMP-primed dialysate adenosine concentration. Accumulation of norepinephrine in the extracellular fluid elicited by pargyline (100 microM), a monoamine oxidase inhibitor, significantly increased histamine-induced adenosine production. Okadaic acid (50 microM), an inhibitor of protein phosphatase, enhanced the histamine-induced increase in adenosine concentration. Norepinephrine is known to activate alpha1-adrenoceptors and PKC. Taken together, the results demonstrate that histamine-released norepinephrine activates both alpha1-adrenoceptors and PKC, which increased ecto-5'-nucleotidase activity and augmented release of adenosine in rat hearts. PMID- 11408528 TI - Bradykinin B1 receptor up-regulation by interleukin-1beta and B1 agonist occurs through independent and synergistic intracellular signaling mechanisms in human lung fibroblasts. AB - Bradykinin B1 receptors (B1R) are rapidly induced after tissue trauma and are thought to be involved in maintaining the inflammatory response. Little is known about the intracellular signaling pathways mediating B1R induction in response to stress and inflammation. Here, we show that up-regulation of B1R by B1R agonist and interleukin-1beta (IL-1beta) occur through distinct but synergistic pathways in IMR-90 human lung fibroblasts. Incubation of cells with the B1R agonist desArg10kallidin (desArg10KD; 100 nM) and IL-1beta (500 pg/ml) resulted in a 3- and 4-fold increase, respectively, in B1R by 6 h, whereas coincubation of these factors produced up to a 20-fold increase. Furthermore, coincubation increased the potency of IL-1beta by 2-fold. Both the individual and the synergistic responses were sensitive to genistein, a general tyrosine kinase inhibitor. On the other hand, only the desArg10KD response and the synergistic response were sensitive to the p38 mitogen-activated protein kinase inhibitor SB 203580. Furthermore, only the synergistic response was sensitive to the nuclear factor kappaB inhibitor pyrrolidine dithiocarbamate. Despite B1R up-regulation in A549 human lung epithelial cells by desArg10KD or IL-1beta individually, these factors did not act synergistically in this cell line. In conclusion, our results reinforce the view that kinins act in concert with proinflammatory cytokines to enhance selectively the inflammatory response of certain lung cells to kinins through distinct but synergistic intracellular signaling mechanisms. Thus, kinins may exert a pivotal role in maintaining and modulating feed-forward inflammatory processes in the lung. PMID- 11408529 TI - Positive modulation of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors in prefrontal cortical pyramidal neurons by a novel allosteric potentiator. AB - Positive modulators of glutamate alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptors can enhance cognitive function in several species. The present experiments compared the actions of a novel biarylpropylsulfonamide compound, LY404187, with the prototypical benzoylpiperidine, 1-(quinoxalin-6-ylcarbonyl)-piperidine (CX516), on AMPA receptors of prefrontal cortex (PFC) pyramidal neurons. LY404187 (0.03-10 microM) selectively enhanced glutamate-evoked currents through AMPA receptor/channels of acutely isolated pyramidal neurons with considerably greater potency (EC50 = 1.3 +/- 0.3 microM) and efficacy (Emax = 45.3 +/- 8.0-fold increase) than did CX516 (EC50 = 2.8 +/- 0.9 mM; Emax = 4.8 +/- 1.4-fold increase). Both LY404187 and CX516 increased the potency of the glutamate concentration-response profile by 6- and 3-fold, respectively. Rapid perfusion experiments demonstrated that LY404187 produced a marked suppression in the magnitude but no change in the kinetics of receptor desensitization; whereas CX516 produced little change in the degree and a modest deceleration of the desensitization process. In PFC slices, both spontaneous and stimulus-evoked AMPA receptor-mediated excitatory postsynaptic potentials were enhanced by nanomolar concentrations of LY404187. Voltage sensitive N-methyl-D-aspartate (NMDA) receptor-dependent synaptic responses also were indirectly augmented as a consequence of greater postsynaptic depolarization. Consistent with the in vitro data, LY404187 was 1000-fold more potent than CX516 in enhancing the probability of discharge of PFC neurons in response to stimulation of glutamatergic afferents from hippocampus in vivo. This potentiation by LY404187 was reduced by both selective AMPA (LY300168, 1 mg/kg, i.v.) and NMDA (LY235959, 5 mg/kg, i.v.) receptor antagonists. Collectively, these results demonstrate that LY404187 is an extremely potent and centrally active potentiator of native AMPA receptors and has a unique mechanism of action. The therapeutic implications of AMPA receptor potentiators are discussed. PMID- 11408530 TI - D609 inhibits ionizing radiation-induced oxidative damage by acting as a potent antioxidant. AB - Tricyclodecan-9-yl-xanthogenate (D609) has been extensively studied in biological systems and exhibits a variety of biological functions, including antiviral, antitumor, and anti-inflammatory activities. Most of these activities have been largely attributed to the inhibitory effect of D609 on phosphatidylcholine specific phospholipase C. However, as a xanthate derivative, D609 is a strong electrolyte and readily dissociates to xanthate anions and cations of alkali metals in solution. Xanthate anions and protonated xanthic acid contain a free thiol moiety and are highly reductive. This implies that D609 and other xanthate derivatives may function as potent antioxidants. Indeed, we found that D609 inhibited the Fenton reaction-induced oxidation of dihydrorhodamine 123 in a dose dependent manner similar to that of pyrrolidinedithiocarbamate, a well known antioxidant. In addition, D609 inhibited the formation of the alpha-phenyl-tert butylnitrone-free radical spin adducts and lipid peroxidation of synaptosomal membranes by the Fenton reagents. Furthermore, preincubation of lymphocytes with D609 resulted in a significant diminution of ionizing radiation (IR)-induced 1) production of reactive oxygen species; 2) decrease in intracellular reduced glutathione; 3) oxidative damage to proteins and lipids; and 4) activation of nuclear factor-kappaB. Moreover, when D609 (50 mg/kg i.v.) was administered to mice 10 min prior to total body IR (6.5 and 8.5 Gy), it protected the mice from IR-induced lethality. Thus, these results indicate that D609 is a potent antioxidant and has the ability to inhibit IR-induced cellular oxidative stress. PMID- 11408531 TI - Comparison of "type I" and "type II" organic cation transport by organic cation transporters and organic anion-transporting polypeptides. AB - Previous inhibition studies with taurocholate and cardiac glycosides suggested the presence of separate uptake systems for small "type I" (system1) and for bulky "type II" (system2) organic cations in rat hepatocytes. To identify the transport systems involved in type I and type II organic cation uptake, we compared the organic cation transport properties of the rat and human organic cation transporter 1 (rOCT1; hOCT1) and of the organic anion-transporting polypeptides 2 and A (rat Oatp2; human OATP-A) in cRNA-injected Xenopus laevis oocytes. Based on characteristic cis-inhibition patterns of rOCT1-mediated tributylmethylammonium and Oatp2-mediated rocuronium uptake, rOCT1 and Oatp2 could be identified as the organic cation uptake systems1 and 2, respectively, in rat liver. While hOCT1 exhibited similar transport properties as rOCT1, OATP-A- but not Oatp2-mediated rocuronium uptake was inhibited by the OATP-A substrate N methyl-quinidine. The latter substrate was also transported by rOCT1 and hOCT1, demonstrating distinct organic cation transport activities for rOCT1 and Oatp2 and overlapping organic cation transport activities for hOCT1 and OATP-A. Finally, the data demonstrate that unmethylated quinidine is transported by rOCT1, hOCT1, and OATP-A at pH 6.0, but not at pH 7.5, indicating that quinidine requires a positive charge for carrier-mediated uptake into hepatocytes. In conclusion, the studies demonstrate that in rat liver the suggested organic cation uptake systems1 and 2 correspond to rOCT1 and Oatp2, respectively. However, the rat-based type I and II organic cation transporter classification cannot be extended without modification from rat to human. PMID- 11408532 TI - Stimulation of guanosine-5'-o-(3-[35S]thio)triphosphate binding in digitonin permeabilized C6 rat glioma cells: evidence for an organized association of mu opioid receptors and G protein. AB - The guanosine-5'-O-(3-[35S]thio)triphosphate ([35S]GTPgammaS) binding assay for the determination of relative opioid efficacy has been adapted to measure G protein activation in digitonin-permeabilized C6 rat glioma cells expressing a cloned mu-opioid receptor. The mu-agonist [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin (DAMGO) caused a 3-fold increase in [35S]GTPgammaS binding over basal in a naloxone-sensitive manner. Relative mu-agonist efficacy was DAMGO > fentanyl > or = morphine > buprenorphine. Nalbuphine showed no efficacy. G protein activation by receptors has been predicted to occur by random encounter. In this model a reduction in the number of receptors will decrease the rate of G protein activation but not the maximum number of G proteins activated. To test this model C6 mu cells were treated with the irreversible mu-antagonist beta-funaltrexamine (10 nM) prior to permeabilization. This reduced the number of mu-opioid receptors determined with [3H]diprenorphine to 23 +/- 3% of control with no change in affinity. A commensurate reduction (to 29 +/- 10% of control) in the level of [35S]GTPgammaS binding stimulated by DAMGO was observed, but the t(1/2) for [35S]GTPgammaS binding remained unchanged. Thus, random encounters of receptor and G protein failed to occur in this permeabilized cell preparation. A model that assumes an organized association of G proteins with receptors better describes the activation of G proteins by opioid mu-receptors. PMID- 11408533 TI - Presynaptic alpha7-nicotinic acetylcholine receptors mediate nicotine-induced nitric oxidergic neurogenic vasodilation in porcine basilar arteries. AB - We previously reported that nicotine-induced nitric oxide (NO)-mediated neurogenic vasodilation in the porcine basilar artery was dependent on intact sympathetic innervation. We further demonstrated in this artery that nicotine acted on nicotinic acetylcholine receptors (nAChRs) on presynaptic sympathetic nerve terminals to release norepinephrine (NE), which then acted on beta2 adrenoceptors located on the neighboring NOergic nerve terminals to release NO, resulting in vasodilation. The nature of the nAChRs has not been determined. The nAChR subtype mediating nicotine-induced dilation in isolated porcine basilar arterial rings denuded of endothelium was therefore examined pharmacologically and immunohistochemically. Results from using an in vitro tissue bath technique indicated that relaxation induced by nicotine (100 microM) was blocked by preferential alpha7-nAChR antagonists (methyllycaconitine and alpha-bungarotoxin) and nonspecific nAChR antagonist (mecamylamine) in a concentration-dependent manner, but was not affected by dihydro-beta-erythroidine (a preferential alpha4 nAChR antagonist). These nAChR antagonists did not affect relaxation elicited by transmural nerve stimulation (8 Hz) or that by sodium nitroprusside and NE. Results from double-labeling immunohistochemical studies in whole-mount porcine basilar and middle cerebral arteries and in cultured porcine superior cervical ganglia (SCG) indicated that alpha7-nAChR- and tyrosine hydroxylase immunoreactivities were colocalized in same nerve fibers. These results suggest the presence of functional alpha7-nAChRs on postganglionic sympathetic adrenergic nerve terminals of SCG origin, which mediate nicotine-induced neurogenic NOergic vasodilation. These findings are consistent with our hypothesis that nicotine acts on nAChRs on presynaptic sympathetic nerve terminals to release NE, which then acts on presynaptic beta2-adrenoceptors located on the neighboring NOergic nerve terminals, resulting in release of NO and dilation of porcine basilar arteries. PMID- 11408534 TI - Dopamine D1 and D2 receptors influence dopamine transporter synthesis and degradation in the rat. AB - Neurotransmitter transporters play an important role in maintaining synaptic homeostasis and in the actions of many drugs. Utilizing a technique for measuring the kinetics (synthesis, degradation, and half-life) of the dopamine transporter (DAT) protein in the rat striatum and nucleus accumbens, we have investigated the effects of systemic administration of dopamine receptor agonists and antagonists upon DAT kinetics in these brain regions. In the striatum, the dopamine D1 receptor agonist SKF38393 and the dopamine D1 receptor antagonist SCH23390 were without effect. However, the dopamine D2 receptor agonists R-(-) propylnorapomorphine hydrochloride (NPA) and quinpirole decreased the half-life of DAT. This effect was blocked by the dopamine D2 antagonist eticlopride, which, by itself, increased the half-life of DAT. In the nucleus accumbens, the agonist SKF38393 increased the DAT half-life, whereas the antagonist SCH23390 decreased the half-life. In contrast to the striatum, NPA and quinpirole increased the DAT half-life, which was blocked by eticlopride and by itself had no effect on DAT kinetics. Cocaine increased the half-life of DAT in both the striatum and the nucleus accumbens. The results of the present study suggest that, through dopamine receptors, dopamine indirectly influences DAT protein turnover in the striatum and in the nucleus accumbens, but in different ways. PMID- 11408535 TI - Modulation of 1,3-bis-(2-chloroethyl)-1-nitrosourea resistance in human tumor cells using hammerhead ribozymes designed to degrade O6-methylguanine DNA methyltransferase mRNA. AB - O6-Methylguanine DNA Methyltransferase (MGMT) protects tumor cells from the cytotoxic effects of the DNA alkylating agent 1,3-bis-(2-chloroethyl)-1 nitrosourea (BCNU). To improve the therapeutic index of BCNU, biochemical strategies to deplete MGMT activity have been developed. In the present study, a molecular strategy for modulating BCNU resistance was explored using hammerhead ribozymes (Rz) designed to degrade the long-lived MGMT mRNA. The ribozymes were designed against eight GUC sites within the MGMT mRNA. cDNAs of these ribozymes were cloned into an expression vector and then all eight vectors were pooled and stably transfected into HeLa cells. Several HeLa/Rz clones sensitive to a sublethal dose of BCNU were identified using a short-term cell proliferation assay. The ribozyme inserts were amplified from genomic DNA by polymerase chain reaction and sequenced in the BCNU-sensitive clones. The ribozyme inserts Rz161, 178, and 212, targeted against nucleotide 161, 178, and 212, respectively, in the MGMT mRNA, were found to be present in these clones. MGMT activity, Western, and Northern blot analyses revealed that two of the HeLa/Rz clones contained very low levels of MGMT activity, protein, and mRNA. Investigation of CpG methylation within the MGMT promoter indicated that the lack of MGMT expression in these HeLa/Rz clones was not likely due to methylation silencing of the MGMT gene. By colony formation, the cell killing induced by 100 microM BCNU was increased by 2 to 3 logs in the HeLa/Rz clones compared with wild-type HeLa cells. PMID- 11408536 TI - Strain-specific effects of amphetamine on prepulse inhibition and patterns of locomotor behavior in mice. AB - Several reports describe substantive behavioral differences between strains of mice both at baseline and in response to pharmacological manipulations. For example, mouse strain differences have been reported in prepulse inhibition (PPI) and patterns of locomotor activity, two behavioral processes that are altered by dopamine (DA) agonists such as amphetamine. Here, we characterized acoustic and tactile startle reactivity, acoustic PPI, and both the amounts and spatial patterns of locomotor activity in C57BL/6J, 129SvEv (129S6), and 129SvJ (129X1) mice at baseline and in amphetamine dose-response studies. Because hearing loss is common in numerous strains of mice, we also assessed cross-modal PPI using a light prepulse with an airpuff startle stimulus. The results establish that these three inbred strains of mice display both intra- and cross-modal PPI, and that amphetamine decreases PPI and startle reactivity in a dose-, sensory modality-, and strain-specific manner. Furthermore, the amount of locomotor activity and the spatial pattern of motor sequences are altered differentially after treatment with amphetamine in C57BL/6J and 129X1 mice, but not in 129S6 mice. Given that amphetamine releases presynaptic DA, these findings are consistent with the role of DA in the modulation of PPI and motor patterns in mice. These findings highlight the importance of selecting appropriate strains of mice for behavioral, pharmacological, and genetic studies. PMID- 11408537 TI - Acute opioid pretreatment potentiates naltrexone-induced drinking suppression in water-deprived rats. AB - Pretreatment with morphine-like agonists potentiates the behavioral effects of opioid antagonists, possibly reflecting a state of acute physical dependence. Several studies have used operant behavior to quantify these effects. However, little research has been done using unconditioned behavior. One objective of this study was to determine whether opioid agonist pretreatment (e.g., morphine, fentanyl, and meperidine) potentiated naltrexone-induced suppression of water consumption following deprivation. Another objective was to determine whether the agonist pretreatment interval was functionally related to efficacy for the manifestation of acute dependence. Finally, we compared temporally the effects of the three agonists. Adult male Sprague-Dawley rats were water deprived for 18, 20, or 22 h and given an injection (s.c.) of an agonist or saline. After 1.75, 3.75, or 5.75 h, animals received a single dose (s.c.) of naltrexone (0.01-30 mg/kg) or saline. Fifteen minutes later, subjects had access to water for 30 min. A time course of antinociception was constructed after agonist administration, using the tail-flick procedure. All three agonists dose dependently potentiated naltrexone-induced drinking suppression, decreasing the ED50 of naltrexone by as much as 150-fold. There was no clear relationship between agonist efficacy and pretreatment interval. Sensitization to naltrexone was seen up to 6 h after agonist administration, occurring in the apparent absence of an antinociceptive effect. These data extend the range of behavioral effects of opioid antagonists potentiated by opioid agonist pretreatment to suppression of drinking and show that such potentiation can occur in the absence of a prototypical agonist effect. PMID- 11408538 TI - Pharmacokinetics, pharmacodynamics, and platelet binding of an anti-glycoprotein IIb/IIIa monoclonal antibody (7E3) in the rat: a quantitative rat model of immune thrombocytopenic purpura. AB - The pharmacokinetics, pharmacodynamics, and platelet binding of 7E3, an anti glycoprotein IIb/IIIa (GPIIb/IIIa) monoclonal antibody, were studied in the rat in an attempt to develop a quantitative animal model of immune thrombocytopenia (ITP). 7E3, a murine IgG1 antibody developed against human GPIIb/IIIa, demonstrated cross-reactivity with rat platelets by flow cytometry and via enzyme linked immunosorbent assay. The apparent affinity (K(A)) of 7E3-rat platelet binding was 1.2 +/- 0.2 x 10(7) M(-1), with 3.3 +/- 0.3 x 10(4) binding sites per platelet. Following intravenous 7E3 administration (0.8, 4, and 8 mg/kg), plasma concentrations declined in a bi-exponential manner, with a terminal half-life of 61 +/- 5 h and a steady-state volume of distribution of 62 +/- 15 ml/kg. Clearance was dose-dependent, with values ranging from 0.64 +/- 0.08 ml/h/kg (8 mg/kg) to 1.01 +/- 0.08 ml/h/kg (0.8 mg/kg). 7E3 induced a reproducible, severe thrombocytopenia in rats and extended bleeding in a manner consistent with human ITP. Nadir platelet counts were 79 +/- 33, 25 +/- 6, and 17 +/- 2 x 10(6)/ml, for 7E3 doses of 0.8, 4, and 8 mg/kg, respectively. Bleeding times after a 10-mm tail incision ranged from 5 +/- 3 min in control animals to 15 +/- 0 min (the maximum allowed time in this study) in animals receiving 8 mg/kg. Blood volumes lost during bleeding experiments ranged from 30 +/- 24 microl (control) to 349 +/- 358 microl (8 mg/kg). A reproducible, quantitative rat model of ITP has been created; this model is expected to facilitate the evaluation of new treatments for this disease. PMID- 11408539 TI - Lobeline attenuates d-methamphetamine self-administration in rats. AB - alpha-Lobeline inhibits d-amphetamine-evoked dopamine release from striatal slices in vitro, appearing to reduce the cytosolic pool of dopamine available for reverse transport by the dopamine transporter. Based on this neurochemical mechanism of action, the present study determined if lobeline decreases d methamphetamine self-administration. Rats were surgically implanted with jugular catheters and were trained to lever press on a fixed ratio 5 schedule for intravenous d-methamphetamine (0.05 mg/kg/infusion). To assess the specificity of the effect of lobeline, another group of rats was trained to lever press on a fixed ratio 5 schedule for sucrose reinforcement. Pretreatment of rats with lobeline (0.3-3.0 mg/kg, 15 min prior to the session) decreased responding for both d-methamphetamine and sucrose reinforcement. Following repeated lobeline (3.0 mg/kg) administration, tolerance developed to the decrease in responding for sucrose; however, the lobeline-induced decrease in responding for d methamphetamine persisted. Furthermore, the lobeline-induced decrease in responding for d-methamphetamine was not surmounted by increasing the unit dose of d-methamphetamine. These results suggest that lobeline produces a nonspecific rate suppressant effect following acute administration, to which tolerance develops following repeated administration. Importantly, the results also suggest that repeated administration of lobeline specifically decreases responding for d methamphetamine in a noncompetitive manner. Thus, lobeline may be an effective, novel pharmacotherapy for d-methamphetamine abuse. PMID- 11408540 TI - Cocaine-mediated apoptosis in bovine coronary artery endothelial cells: role of nitric oxide. AB - The present study examined the role of nitric oxide in cocaine-induced apoptosis in bovine coronary artery endothelial cells (BCAECs). Cocaine produced a time dependent decrease in cell viability and an increase in apoptosis in BCAECs, which were blocked by the nitric oxide donors DETA-NONOate (DETA-NO) and S nitroso-N-acetyl-penicillamine. In accordance, cocaine decreased nitric oxide production in BCAECs at each time point of the study. Cocaine significantly increased caspase-3 activity that was blocked by the inhibitors of cytochrome c release (cyclosporin A), caspase-3 (Ac-DEVD-CHO), and caspase-9 (Z-LEHD-FMK), respectively. In addition, cocaine activated caspase-9, which was blocked by cyclosporin A and Z-LEHD-FMK. Ac-DEVD-CHO only partially blocked cocaine-induced caspase-9 activity. DETA-NO (20 microM) blocked cocaine-mediated activation of both caspase-9 and caspase-3. Cocaine decreased Bcl-2 protein levels, which was partially blocked by Ac-DEVD-CHO and Z-LEHD-FMK, but not by DETA-NO. Furthermore, cocaine induced a translocation of Bax from the cytosol to the mitochondria in BCAECs, and increased Bax levels in mitochondria by 2.2-fold. In accordance, cytosolic Bax levels decreased about 42%. Neither Ac-DEVD-CHO nor DETA-NO affected cocaine-induced translocation of Bax. We conclude that cocaine-induced Bcl-2 protein down-regulation and Bax translocation to the mitochondria are upstream signals of caspase-9 activation that precedes caspase-3. Cocaine-induced attenuation of nitric oxide plays a key role in the activation of the caspase cascade in BCAECs. PMID- 11408541 TI - Enhanced cardiac L-type calcium current response to beta2-adrenergic stimulation in heart failure. AB - The beta2-adrenergic receptor (beta2-AR)-mediated increase in cardiac L-type Ca2+ current (I(Ca,L)) has been documented in normal subjects. However, the role and mechanism of beta2-AR activation on I(Ca,L) in heart failure (HF) are unclear. Accordingly, we compared the effect of zinterol (ZIN), a highly selective beta2 AR agonist, on I(Ca,L) in isolated left ventricular cardiomyocytes obtained from normal control and age-matched rats with HF induced by left coronary artery ligation (4 months). I(Ca,L) was measured by using the whole-cell voltage-clamp technique. In normal myocytes, superfusion of ZIN (10(-5) M) caused a 21% increase in I(Ca,L) (9.21 +/- 0.24 versus 7.59 +/- 0.20 pA/pF) (p < 0.05). In HF myocytes, the same concentration of ZIN produced a significantly greater increase (30%) in I(Ca,L) (6.20 +/- 0.24 versus 4.75 +/- 0.17 pA/pF) (p < 0.01). This ZIN induced increase in I(Ca,L) was further augmented in both normal and HF myocytes (normal: 59 versus 21%; HF: 71 versus 30%) after the incubation of myocytes with pertussis toxin (PTX, 2 microg/ml, 36 degrees C, 6 h). These effects were not modified by the incubation of myocytes with CGP-20712A (3 x 10(-7) M), a beta1-AR antagonist, but were abolished by pretreatment of myocytes with ICI-118551 (10( 7) M), a beta2-AR antagonist. In addition, all of the effects induced by ZIN were completely prevented in the presence of an inhibitory cAMP analog, Rp-cAMPS (100 microM, in the patch-pipette solution). In conclusion, beta2-AR activation stimulates L-type Ca2+ channels and increases I(Ca,L) in both normal and HF myocytes. In HF, beta2-AR activation-induced augmentation of I(Ca,L) was increased. These effects are likely to be mediated through a cAMP-dependent mechanism and coupled with both stimulatory G protein and PTX-sensitive G protein. PMID- 11408542 TI - Geraniol, a component of plant essential oils, inhibits growth and polyamine biosynthesis in human colon cancer cells. AB - Geraniol and other monoterpenes found in essential oils of fruits and herbs have been suggested to represent a new class of agents for cancer chemoprevention. As a first step in clarifying the mode of action of geraniol on colon carcinogenesis, we studied its effects on the growth of a human colon cancer cell line (Caco-2). Geraniol (400 microM) caused a 70% inhibition of cell growth, with cells accumulating in the S transition phase of the cell cycle, and concomitant inhibition of DNA synthesis. No signs of cytotoxicity or apoptosis were detected. Geraniol caused a 50% decrease of ornithine decarboxylase activity, a key enzyme of polyamine biosynthesis, which is enhanced in cancer growth. This led to a 40% reduction of the intracellular pool of putrescine. Geraniol also activated the intracellular catabolism of polyamines, indicated by enhanced polyamine acetylation. These observations indicate that polyamine metabolism is presumably a target in the antiproliferative properties of geraniol. PMID- 11408543 TI - Coadministered nitrous oxide enhances the effect of isoflurane on GABAergic transmission by an increase in open-channel block. AB - Clinically relevant concentrations of isoflurane (ISO) and nitrous oxide (N2O) enhance chloride currents induced by activating gamma-aminobutyric acid(A) receptors (GABA(A)R). Channel blocking by ISO overcomes the enhancing effect at higher concentrations. In this study, the effect of coadministered ISO and N2O on responses evoked by GABA in transfected human embryonic kidney 293 cells carrying alpha1beta2gamma2L GABA(A)R was investigated. Patch-clamp recordings from these cells were performed in the whole cell mode. A piezo-driven "liquid filament" drug application system was used to apply solutions of GABA, ISO, and N2O. Increasing the concentration of ISO in steps from 0.15 to 1.2 mM resulted in a bell-shaped concentration-response curve for GABA-induced currents. The maximum increase in current (1.51 +/- 0.14-fold) was seen at 0.45 mM ISO (about 1 minimum alveolar concentration, EC50). N2O (29.2 mM) increased GABA-evoked currents 1.54 +/- 0.10-fold. The enhancing effects of ISO and N2O on the GABAergic response were not additive. However, a transient current, associated with the rapid withdrawal of ISO from the receptor, was markedly increased by N2O. Such rebound currents probably reflect the transition from a "channel-blocked" to a "reopened" state. An open-channel block at ligand-gated receptors can prolong postsynaptic currents. Thus, we conclude that coadministered N2O could increase the enhancing effect of ISO on the GABAergic transmission by an increase in open-channel block at the GABA(A)R. PMID- 11408544 TI - Novel short-acting A2A adenosine receptor agonists for coronary vasodilation: inverse relationship between affinity and duration of action of A2A agonists. AB - Several potent and selective A2A adenosine receptor agonists are currently available. These compounds have a high affinity for the A2A receptor and a long duration of action. However, in situations where a short duration of action is desired, currently available A2A receptor agonists are less than ideal. From a series of recently synthesized A2A receptor agonists, two agonists (CVT-3146 and CVT-3033) with low affinity were selected for further characterization as selective and short-acting coronary vasodilators. Both compounds were selective for the A2A adenosine receptor (AdoR) versus the A1, A2B, and A3AdoR in binding and functional studies. CVT-3146 and CVT-3033 appeared to be weak partial agonists to cause cAMP accumulation in PC12 cells, but were full and potent agonists to cause coronary vasodilation, a response that has a very large A2A receptor reserve. However, the durations of action of CVT-3146 and CVT-3033 were remarkably shorter than those of the high-affinity agonists CGS21680 or WRC0470, presumably due to the relative lower affinity of CVT-3146 and CVT-3033 for the A2A receptor. Indeed, an inverse relationship was found between the affinity of the various agonists for the A2A receptor and the duration of their actions. These data indicate that low-affinity agonists can produce a response that is of equivalent magnitude but more rapid in termination than that caused by a high affinity agonist. Hence, the low-affinity A2A agonists CVT-3146 and CVT-3033 may prove to be superior to currently available high-affinity agonists as coronary vasodilators during myocardial imaging with radionuclide agents. PMID- 11408545 TI - Rabbit alpha2-adrenoceptors: both platelets and adipocytes have alpha2A pharmacology. AB - The recombinant alpha2-adrenoceptors, designated as alpha2a and alpha2d, have highly similar amino acid sequences, but distinct pharmacological properties. It has been suggested that these two receptor subtypes are species orthologs, since the alpha2-adrenoceptors of a given species have pharmacological characteristics corresponding to either the alpha2a- (human, pig) or alpha2d- (rat, mouse, guinea pig, cow) adrenoceptor. Radioligand binding assays in rabbit adipocyte suggest alpha2D-adrenoceptor pharmacology. However, functional studies examining prejunctional alpha2-adrenoceptors in several tissues pharmacologically define the receptor of the rabbit as an alpha2A-adrenoceptor rather than an alpha2D adrenoceptor. We characterized the alpha2-adrenoceptor of rabbit adipocyte and platelet, comparing the ability of norepinephrine and 13 adrenoceptor antagonists to inhibit the binding of [3H]RX821002 with the affinity of these drugs for the human alpha2a-adrenoceptor or the rat alpha2d-adrenoceptor. Pharmacological characteristics of the adipocyte and platelet receptor were very similar, with an excellent correlation between pK(i) values (r2 = 0.95, slope of regression = 1.01). Drug affinities for both platelet and adipocyte receptors correlated better with the alpha2a-adrenoceptor (r2 = 0.68-0.77) than with the alpha2d adrenoceptor (r2 = 0.37-0.38). Despite the relatively low affinity of the rabbit adipocyte alpha2-adrenoceptor for yohimbine and rauwolscine, this receptor, as well as the platelet receptor, have alpha2A-adrenoceptor pharmacology. Subtle differences in the alpha2-adrenoceptor binding characteristics of these native rabbit tissues compared with the recombinant human alpha2a-adrenoceptor may result either from minor differences in the sequence of human and rabbit alpha2a adrenoceptors or from differences in the environment to which native and recombinant receptors are exposed. PMID- 11408546 TI - Dopamine-independent locomotion following blockade of N-methyl-D-aspartate receptors in the ventral tegmental area. AB - Compounds acting in the ventral tegmental area to increase motor activity are thought to do so by activating mesolimbic dopamine transmission. The present report demonstrates that the microinjection of N-methyl-D-aspartate (NMDA) antagonists into the ventral tegmental area produces a dose-dependent increase in motor activity. This effect was not mimicked by antagonizing either alpha-amino-3 hydroxy-5-methyl-4-isoxazolepropionic acid/kainate or metabotropic glutamate receptors in the ventral tegmental area. Three experiments were conducted that indicated that the capacity of NMDA receptor antagonists to elevate motor activity did not involve increased dopamine transmission. 1) The systemic administration of a D1 dopamine receptor antagonist did not inhibit the motor stimulant response to NMDA antagonist injection into the ventral tegmental area except at doses that also inhibited motor activity after an injection of saline into the ventral tegmental area. 2) Stimulating orphanin receptors in the ventral tegmental area selectively inhibits dopamine cells, and this did not alter NMDA antagonist-induced motor activity. Whereas, stimulating gamma-aminobutyric acid (GABA)(B) receptors hyperpolarizes both dopamine and GABA cells in the ventral tegmental area, and this abolished NMDA antagonist-induced motor activity. 3) The microinjection of an NMDA antagonist into the ventral tegmental area did not increase dopamine metabolism in dopamine terminal fields, including the accumbens, striatum, or prefrontal cortex. Also consistent with a lack of dopamine involvement, repeated administration of NMDA antagonist into the ventral tegmental area did not produce behavioral sensitization. These data identify a mechanism to elicit a motor stimulant response from the ventral tegmental area that does not involve activating dopamine transmission. PMID- 11408547 TI - Arsenic induces expression of the multidrug resistance-associated protein 2 (MRP2) gene in primary rat and human hepatocytes. AB - Metals, such as arsenic or cadmium, have recently been demonstrated to interact with metabolic pathways, including phase I and phase II enzymes and the phase III efflux pump P-glycoprotein. In the present study, we investigated the effects of heavy metals and metalloids on the expression of the multidrug resistance associated protein 2 (MRP2), a major hepatic transporter. Treatment of primary rat hepatocytes by sodium arsenite [As(III)], sodium arsenate and potassium antimony tartrate, but not cadmium chloride, was shown to markedly increase MRP2 mRNA and protein levels; As(III)-mediated induction was dose- and time-dependent and paralleled a strong increase in MRP2 amounts as assessed by Western blotting. As(III) was also demonstrated to markedly up-regulate MRP2 gene expression in primary human hepatocytes. MRP2 mRNA induction occurring in As(III)-treated rat hepatocytes was fully blocked by actinomycin D, indicating that it required active gene transcription. It was associated with an activation of the c-Jun N terminal kinase pathway and with a reduction of cellular glutathione levels. Quercetin, a flavonoid compound known to block As(III)-related induction of P glycoprotein, was also found to prevent up-regulation of MRP2 gene expression in rat hepatocytes exposed to As(III). Such an effect was unlikely to be due to alteration of JNK pathway since quercetin failed to abolish As(III)-induced JNK phosphorylation. It may rather be linked to the increase of cellular glutathione levels by quercetin, thus limiting the depleting effects of As(III) on glutathione amounts. Finally, these results confirm that some metals strongly regulate expression of detoxifying proteins, including biliary drug transporters. PMID- 11408548 TI - Agonist Activity of the delta-antagonists TIPP and TIPP-psi in cellular models expressing endogenous or transfected delta-opioid receptors. AB - A new class of highly selective delta-opioid receptor antagonists has been recently developed, termed the TIP(P) peptides. Two prototypical compounds in this class are TIPP (H-Tyr-Tic-Phe-Phe-OH) and a derivative, TIPP-psi (H-Tyr Tic[CH2NH]-Phe-Phe-OH). Surprisingly, both TIPP and TIPP-psi demonstrated inhibition of adenylyl cyclase activity in GH3 cells transfected with delta opioid receptors (GH3DORT), an effect normally observed by agonists. The agonist activity was delta-selective, because no inhibition occurred in wild-type GH3 or GH3MOR (mu-opioid receptor) cells. Both TIPP and TIPP-psi exhibited concentration dependent inhibition of adenylyl cyclase activity; however, TIPP-psi was found to be less potent (IC50 = 3.97 versus 0.162 nM) and less efficacious (I(max) = 50% versus 70%) than TIPP. Pretreatment of cells with pertussis toxin attenuated the inhibition of maximally effective concentrations of TIPP and TIPP-psi, indicating the involvement of G(i)alpha/G(o)alpha G-proteins. Other delta-antagonists, naltriben, naloxone, and ICI 174864, attenuated the inhibition of adenylyl cyclase activity mediated by TIPP. Coadministration of TIPP with the selective delta-agonist [D-Pen2,5]enkephalin resulted in an additive interaction. Both TIPP and TIPP-psi exhibited significant inhibition of adenylyl cyclase activity in different GH3DORT clones expressing a 28-fold range of delta-opioid receptor densities, and in cell lines expressing endogenous (i.e., N1E115 and NG108-15) and transfected (i.e., Chinese hamster ovary-DOR and human embryonic kidney-DOR) delta-opioid receptors, with densities ranging from 0.12 to 6.67 pmol/mg. These results suggest that compounds previously thought to be purely delta-opioid receptor antagonists also demonstrate agonist activity in several in vitro models. PMID- 11408549 TI - SEA0400, a novel and selective inhibitor of the Na+-Ca2+ exchanger, attenuates reperfusion injury in the in vitro and in vivo cerebral ischemic models. AB - The effect of the newly synthesized compound 2-[4-[(2,5 difluorophenyl)methoxy]phenoxy]-5-ethoxyaniline (SEA0400) on the Na+-Ca2+ exchanger (NCX) was investigated and compared against that of 2-[2-[4-(4 nitrobenzyloxy)phenyl]ethyl]isothiourea (KB-R7943). In addition, the effects of SEA0400 on reperfusion injury in vitro and in vivo were examined. SEA0400 was extremely more potent than KB-R7943 in inhibiting Na+-dependent Ca2+ uptake in cultured neurons, astrocytes, and microglia: IC50s of SEA0400 and KB-R7943 were 5 to 33 nM and 2 to 4 microM, respectively. SEA0400 at the concentration range that inhibited NCX exhibited negligible affinities for the Ca2+ channels, Na+ channels, K+ channels, norepinephrine transporter, and 14 receptors, and did not affect the activities of the Na+/H+ exchanger, Na+,K+-ATPase, Ca2+-ATPase, and five enzymes. SEA0400, unlike KB-R7943, did not inhibit the store-operated Ca2+ entry in cultured astrocytes. SEA0400 attenuated dose- dependently paradoxical Ca2+ challenge-induced production of reactive oxygen species, DNA ladder formation, and nuclear condensation in cultured astrocytes, whereas it did not affect thapsigargin-induced cell injury. Furthermore, administration of SEA0400 reduced infarct volumes after a transient middle cerebral artery occlusion in rat cerebral cortex and striatum. These results indicate that SEA0400 is the most potent and selective inhibitor of NCX, and suggest that the compound may exert protective effects on postischemic brain damage. PMID- 11408550 TI - Blockade of opioid receptors in rostral ventral medulla prevents antihyperalgesia produced by transcutaneous electrical nerve stimulation (TENS). AB - Although transcutaneous electrical nerve stimulation (TENS) is used extensively in inflammatory joint conditions such as arthritis, the underlying mechanisms are unclear. This study aims to demonstrate an opiate-mediated activation of descending inhibitory pathways from the rostral ventral medulla (RVM) in the antihyperalgesia produced by low- (4 Hz) or high-frequency (100 Hz) TENS. Paw withdrawal latency to radiant heat, as an index of secondary hyperalgesia, was recorded before and after knee joint inflammation (induced by intra-articular injection of 3% kaolin and carrageenan) and after TENS/no TENS coadministered with naloxone (20 microg/1 microl), naltrindole (5 microg/1 microl), or vehicle (1 microl) microinjected into the RVM. The selectivity of naloxone and naltrindole doses was tested against the mu-opioid receptor agonist [D-Ala2,N-Me Phe4,Gly-ol5]-enkephalin (DAMGO) (20 ng, 1 microl) and the delta2-opioid receptor agonist deltorphin (5 microg, 1 microl) in the RVM. Naloxone microinjection into the RVM blocks the antihyperalgesia produced by low frequency (p < 0.001), but not that produced by high-frequency TENS (p > 0.05). In contrast, naltrindole injection into the RVM blocks the antihyperalgesia produced by high-frequency (p < 0.05), but not low-frequency (p > 0.05) TENS. The analgesia produced by DAMGO and deltorphin is selectively blocked by naloxone (p < 0.05) and naltrindole (p < 0.05), respectively. Thus, the dose of naloxone and naltrindole used in the current study blocks mu- and delta-opioid receptors, respectively. Hence, low frequency and high-frequency TENS produces antihyperalgesia by activation of mu- and delta-opioid receptors, respectively, in the RVM. PMID- 11408551 TI - Riboflavin uptake in human trophoblast-derived BeWo cell monolayers: cellular translocation and regulatory mechanisms. AB - Riboflavin (vitamin B2) is essential for fetal development and must be acquired from maternal sources. The uptake mechanism of riboflavin and the major regulatory pathways involved were characterized in a model for the placental barrier, the human choriocarcinoma cell line, BeWo. Uptake of [3H]riboflavin was saturable (Kt = 1.32 +/- 0.68 nM, Jmax = 266.63 +/- 26.89 fmol/mg of protein/20 min), and was significantly reduced at low temperature and in the presence of metabolic inhibitors (azide, 2-deoxyglucose) or structural analogs. Ouabain, amiloride, sodium-free buffers, and medium with pH values ranging from 3 to 8 did not affect uptake of [3H]riboflavin. In contrast, substitution of chloride with other monovalent anions significantly inhibited its uptake. Induced differentiation of BeWo cells into syncytiotrophoblasts by forskolin or 8-bromo cyclic adenosine monophosphate introduced a time-dependent decrease of riboflavin uptake. Preincubation with activators of cyclic nucleotide-dependent protein kinase pathways (3-isobutyl-1-methylxanthine and p-chlorophenylthio-cyclic guanosine monophosphate) and calmodulin antagonists (calmidazolium and W-13) resulted in a concentration-dependent reduction of [3H]riboflavin uptake, whereas specific modulators of protein kinase C pathways did not have significant effects. 3-Isobutyl-1-methylxanthine exerted its regulatory effect on riboflavin uptake via decreasing both Kt and Jmax of the riboflavin uptake process (Kt = 6.32 +/- 1.29 nM, Jmax = 135.57 +/- 10.42 fmol/mg of protein/20 min). In summary, we report the presence of high- affinity riboflavin transporter(s) on the microvillous membrane of BeWo cells that appears to be modulated by cellular cyclic nucleotide levels and calmodulin. PMID- 11408552 TI - Ca2+/calmodulin-dependent protein kinase II and cytosolic phospholipase A2 contribute to mitogenic signaling in myeloblastic leukemia U-937 cells. AB - The signaling mechanisms downstream of growth factor-stimulated proliferation in myeloid leukemia cells have not yet been fully elucidated. Recent evidence suggests that alternate pathways to the mitogen-activated protein kinase cascade are required. We have previously shown that Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) activates cytosolic phospholipase A2 (cPLA2), which is involved in the proliferation of vascular smooth muscle cells. In the present study, the contribution of this pathway was investigated in the proliferation of U-937 myeloid leukemia cells. In U-937 cells, fetal bovine serum (FBS)-induced proliferation was attenuated by CaM kinase II inhibitor KN-93 but not by its inactive analog KN-92. Inhibitors of cPLA2 (methyl arachidonyl fluorophosphonate and arachidonyl trifluoromethyl ketone) also reduced proliferation of U-937 cells. FBS-induced proliferation was also attenuated by cotransfection with cPLA2 antisense oligonucleotides. These results suggest a role for CaM kinase II and cPLA2 in the proliferation of U-937 cells. FBS stimulated CaM kinase II and cPLA2 activities in a time-dependent manner. Moreover, FBS-stimulated phosphorylation and activation of cPLA2 activation was inhibited by KN-93. FBS-stimulated phosphorylation of CaM kinase II was blocked by KN-93 but not by cPLA2 inhibitors, suggesting that CaM kinase II activates cPLA2. The products of phospholipid hydrolysis produced by cPLA2, lysophosphatidylcholine but not arachidonic acid, increased [3H]thymidine incorporation in U-937 cells. These data suggest that exposure of U-937 cells to FBS promotes phosphorylation and activation of CaM kinase II, leading to stimulation of cPLA2 and generation of lysophosphatidylcholine and resultant proliferation of these cells. PMID- 11408553 TI - Phosphatidylcholine association increases the anti-inflammatory and analgesic activity of ibuprofen in acute and chronic rodent models of joint inflammation: relationship to alterations in bioavailability and cyclooxygenase-inhibitory potency. AB - We investigated whether chemical association of phosphatidylcholine (PC) to ibuprofen enhances the anti-inflammatory/analgesic activity of the nonsteroidal anti-inflammatory drug (NSAID) and whether any change in therapeutic action is due to alterations in drug bioavailability and cyclooxygenase (COX) inhibitory activity. Acute/chronic joint inflammation was induced in rats, by injection of Complete Freund's Adjuvant. In the acute study, rats were administered saline, ibuprofen, or PC-ibuprofen (at NSAID doses of 10, 25, and 50 mg/kg), and 2 h later the pain threshold of the affected joint to pressure was measured. PC ibuprofen increased the pain threshold at all NSAID doses, whereas unmodified ibuprofen demonstrated analgesic activity at only the highest dose. In the chronic study, we investigated the effects of saline, PC-ibuprofen, and ibuprofen (administered at 15 and 25 mg/kg/day) on ankle thickness and pain threshold, and demonstrated that PC-ibuprofen had significantly greater anti-inflammatory and analgesic activity than ibuprofen, over a 30- to 60-day period. PC association resulted in reduced uptake (decreased Cmax), a modest increase in the area under the curve, and a longer t(1/2) of ibuprofen. We also demonstrated that PC ibuprofen was a comparable or a more effective inhibitor of both 6-keto prostaglandin F1alpha concentration of fluid collected from tissue in and around the inflamed stifle joint, and COX-2 activity in activated human umbilical vein endothelial cells. In conclusion, we have demonstrated that PC association results in increases in ibuprofen's anti-inflammatory and analgesic activity in rodent models of acute and chronic joint inflammation, and this effect may relate to alterations in drug bioavailability and COX-inhibitory potency. PMID- 11408554 TI - N-methyl-D-aspartate receptor antagonists potentiate the antinociceptive effects of morphine in squirrel monkeys. AB - Data from rodent antinociception models indicate that N-methyl-D-aspartate (NMDA) receptor antagonists do not produce antinociception alone or potentiate morphine antinociception, but do attenuate the development of morphine tolerance. This study examined the antinociceptive effects of the noncompetitive NMDA receptor antagonist dizocilpine, the competitive NMDA receptor antagonist (-)-6 phosphonomethyl-decahydroisoquinoline-3-carboxylic acid (LY235959), and the glycine-site antagonist (+)-(1-hydroxy-3-aminopyrrolidine-2-one) [(+)-HA-966], alone and in combination with morphine in a squirrel monkey titration procedure. In this procedure, shock (delivered to the tail) increased in intensity every 15 s from 0.01 to 2.0 mA in 30 increments. Five lever presses during any given 15-s shock period produced a 15-s shock-free period after which shock resumed at the next lower intensity. Morphine (0.3-3.0 mg/kg i.m.) dose-dependently increased the intensity below which monkeys maintained shock 50% of the time (median shock level; MSL). In contrast, dizocilpine (0.003-0.1 mg/kg i.m.) produced only modest increases in MSL in some monkeys (three of five) at the highest dose tested. Neither LY235959 (0.1-1.0 mg/kg i.m.) or (+)-HA-966 (10-56 mg/kg i.m.) increased MSL in any monkey tested. Dizocilpine, LY235959, and (+)-HA-966, when administered in combination with doses of morphine (1.0 mg/kg, 1.7 mg/kg) that either produced no antinociception or produced very little antinociception, were all found to dose-dependently potentiate the antinociceptive effect of morphine. Importantly, although these NMDA antagonists in combination with morphine produced marked increases in MSL, these combinations did not alter response rate, demonstrating that the potentiation was not due to nonspecific motor effects. PMID- 11408555 TI - Chronic lipopolysaccharide exposure on airway function, cell infiltration, and nitric oxide generation in conscious guinea pigs: effect of rolipram and dexamethasone. AB - This study investigated whether a correlation between airway hyperreactivity (AHR), leukocyte influx, and nitric oxide (NO) existed in guinea pigs chronically exposed to lipopolysaccharide (LPS). The effect of the corticosteroid, dexamethasone, or phosphodiesterase-4 (PDE4) inhibitor, rolipram, on these features was studied. Airway function was measured in conscious guinea pigs (specific airways conductance) before and after single, double, or chronic (nine) LPS (30 microg x ml(-1), 1 h) exposures. Airway reactivity to inhaled histamine (1 mM, 20 s) was assessed before and at various times after LPS challenges. Leukocytes and NO metabolites were measured in bronchoalveolar lavage fluid (BALF). AHR occurred at 1 h after a single LPS challenge and was resolved by 4 h. This coincided with reduction and recovery, respectively, of BALF NO levels. The AHR and NO deficiency were extended to 4 h, after a double LPS exposure. Chronic LPS exposures, 48 h apart, initially caused persistent bronchodilations, whereas later exposures produced progressively persistent bronchoconstrictions. There was AHR 24 h after the eighth challenge. Twenty-four hours after the ninth LPS exposure, macrophages, neutrophils, eosinophils, and NO metabolites were elevated in BALF. Dexamethasone (20 mg x kg(-1) i.p.) or rolipram (1 mg x kg(-1) i.p.) prevented single and chronic LPS-induced AHR, the respective deficiency and elevation in NO metabolites, and the chronic LPS-induced leukocyte influx. Dexamethasone exacerbated, whereas rolipram reversed, the chronic LPS-induced bronchoconstrictions. This study demonstrates for the first time that chronic LPS causes persistent bronchoconstriction, neutrophilic inflammation, AHR, and NO overproduction in guinea pig airways. These rolipram-sensitive features suggest the potential of PDE4 inhibitors in airway disease. PMID- 11408556 TI - Pharmacological characterization of ZD6021: a novel, orally active antagonist of the tachykinin receptors. AB - The tachykinins, substance P, neurokinin A, and neurokinin B, have been implicated in many diseases. The present study evaluated the pharmacological properties of a novel tachykinin antagonist ZD6021 [3-cyano-N-((2S)-2-(3,4 dichlorophenyl)-4-[4-[2-(methyl-(S)-sulfinyl)-phenyl]piperidino]butyl)-N-methyl-] napthamide]. The affinity (K(i)) of ZD6021 for the cloned human neurokinin (NK)1, NK2, and NK3 receptors was 0.12 +/- 0.01, 0.64 +/- 0.08, and 74 +/- 13 nM, respectively. Mucin secretion by Chinese hamster ovary cells transfected with the human NK1 receptor was dose dependently inhibited by ZD6021: pIC(50) = 7.6 +/- 0.1. For NK1 and NK2 receptors, the agonist concentration-response curves using isolated tissues were displaced rightward in the presence of ZD6021: rabbit pulmonary artery, pA2 = 8.7 and 8.5; human pulmonary artery and bronchus, pKB = 8.9 +/- 0.4 and 7.5 +/- 0.2, at 10(-7) M, respectively. Senktide-induced contractions of isolated guinea pig ileum were also blocked by low concentrations of ZD6021. Oral administration of ZD6021 to guinea pigs dose dependently attenuated tracheal extravasation of plasma proteins induced by the NK1 receptor agonist Ac-[Arg6,Sar9,Met(O2)11]-SP(6-11), ED50 = 0.8 micromol/kg, and bronchoconstriction, elicited by the NK2 receptor agonist [beta-Ala8]-NKA(4-10), ED50 = 20 micromol/kg. Potency was unaffected by feeding. After oral administration of ZD6021, the time to peak activity was 150 min for the NK1 receptor and 60 min for the NK2 receptor with pharmacodynamic half-lives of 280 and 458 min, respectively. These data indicate that ZD6021 is a potent, orally active antagonist of all three tachykinin receptors. This compound may be useful for future studies of tachykinin-related pathology such as asthma. PMID- 11408557 TI - Characterization of the efflux transport of 17beta-estradiol-D-17beta-glucuronide from the brain across the blood-brain barrier. AB - The contribution of organic anion transporters to the total efflux of 17beta estradiol-D-17beta-glucuronide (E(2)17betaG) through the blood-brain barrier (BBB) was investigated using the Brain Efflux Index method by examining the inhibitory effects of probenecid, taurocholate (TCA), p-aminohippurate (PAH), and digoxin. E(2)17betaG was eliminated through the BBB with a rate constant of 0.037 min(-1) after the microinjection into the brain. Probenecid and TCA inhibited this elimination with an IC50 value of 34 and 1.8 nmol/0.5 microl of injectate, respectively, whereas PAH and digoxin reduced the total efflux to about 80 and 60% of the control value, respectively. The selectivity of these inhibitors was confirmed by examining their inhibitory effects on the transport via organic anion transporting polypeptide 1 (Oatp1), Oatp2, organic anion transporter 1 (Oat1), and Oat3 transfectants using LLC-PK1 cells as hosts. Digoxin specifically inhibited the transport via Oatp2 (K(i) = 0.037 microM). The K(i) values of TCA for Oatp1 and Oatp2 (11 and 39 microM, respectively) were about 20 times lower than those for Oat1 and Oat3 (2.8 and 0.8 mM, respectively). PAH did not affect the transport via the Oatp family, but had a similar affinity for Oat1 and Oat3 (85 and 300 microM, respectively). Probenecid had a similar affinity for these transporters (Oatp1, Oatp2, Oat1, and Oat3) examined in this study. Taking the selectivity of these inhibitors into consideration, the maximum contribution made by the Oatp2 and Oat family to the total efflux of E(2)17betaG from the brain appears to be about 40 and 20%, respectively. PMID- 11408558 TI - P-glycoprotein-mediated efflux of indinavir metabolites in Caco-2 cells expressing cytochrome P450 3A4. AB - The role of P-glycoprotein in secretion of indinavir metabolites produced by CYP3A4 was evaluated in Caco-2 cells expressing CYP3A4. Metabolism of indinavir by CYP3A4 expressing Caco-2 cells grown on filters resulted in the formation of N dealkylation products (M5 and M6) and hydroxylation of indinavir, which were preferentially secreted into the apical compartment. Apical secretion of the metabolites was inhibited by cyclosporin A (CsA) with all three classes of metabolites showing similar sensitivity to CsA, suggesting that they are all secreted by the same pathway. M6 stimulated P-glycoprotein (Pgp)-ATPase activity in a concentration-dependent manner. This stimulation was inhibited by the Pgp specific monoclonal antibody C219. A method was developed to specifically inhibit Pgp using the monoclonal antibody UIC2 to determine whether Pgp efflux accounts for a significant proportion of the apical secretion of indinavir metabolites. UIC2 recognizes an extracellular transient conformational epitope that is stabilized by some Pgp substrates or by ATP depletion. Incubation of Caco-2 cells with UIC2 in the presence of 1 microM CsA resulted in 50 to 80% inhibition of Pgp mediated vinblastine efflux, with no significant inhibition observed by UIC2 or CsA alone. Inhibition of Pgp in CYP3A4-expressing Caco-2 cells by UIC2 and 1 microM CsA resulted in a significant decrease in the apical secretion of M6, M5, and OH-indinavir and an increase in the amount of the metabolites secreted in the basolateral compartment and retained in the cytosol. These results are consistent with a role of Pgp in elimination of CYP3A4-generated metabolites and indicate that even relatively polar metabolites may be secreted from the cell by Pgp. PMID- 11408559 TI - Cytosolic phospholipase A2 activation by the p38 kinase inhibitor SB203580 in rabbit aortic smooth muscle cells. AB - SB203580 [4-(4-fluorophenyl)-2-(4-methylsulfinylphenyl)-5-(4-pyridyl)1H imidazole] is widely used as a specific inhibitor of p38 mitogen-activated protein kinase (MAPK). Here we report that SB203580, which blocked p38 kinase activation elicited by anisomycin, increased the phosphorylation and activity of cytosolic phospholipase A2 (cPLA2) and arachidonic acid (AA) release in quiescent vascular smooth muscle cells from rabbit aortae. SB203580 also increased the activity of calcium (Ca2+)/camodulin-dependent kinase II (CaMKII) and ERK1/2 MAPK. The increase in CaMKII activity and cPLA2 phosphorylation caused by SB203580 was attenuated by CaMKII inhibitor KN-93, indicating involvement of CaMKII in cPLA2 phosphorylation by this compound. Since KN-93 also inhibited SB203580-induced ERK1/2 activation, it appears that ERK1/2 activation is also mediated by CaMKII. SB203580-induced cPLA2 phosphorylation was inhibited by depletion of Ca2+ from the medium, by the voltage-operated Ca2+ channel blocker nifedipine, and by the calmodulin inhibitor W-7. cPLA2 translocation from cytoplasm to the nuclear envelope caused by SB203580 was also inhibited in the absence of extracellular Ca2+. Other p38 kinase inhibitors, SB202190 and PD169316, failed to alter CaMKII, ERK1/2, and cPLA2 activity or cPLA2 translocation to the nuclear envelope. These data suggest that SB203580 not only inhibits p38 kinase activity but also increases Ca2+ influx through voltage sensitive Ca2+ channels, which promotes cPLA2 translocation to the nuclear envelope, and by interacting with calmodulin, activates CaMKII and cPLA2 and releases AA. PMID- 11408560 TI - Pharmacologic or genetic manipulation of glutathione S-transferase P1-1 (GSTpi) influences cell proliferation pathways. AB - Glutathione S-transferase P1-1 (GSTpi) is an abundant and ubiquitously expressed protein in normal and malignant mammalian tissues and possesses catalytic and ligand binding properties. Our present data suggest that the protein contributes to the regulation of cell proliferation. Mouse embryo fibroblasts (MEFs) isolated from mice with a GSTP1-1 [glutathione S-transferase P1-1 (isozyme in nonhepatic tissue)] null genotype (GSTpi(-/-)) doubled their population in 26.2 h versus 33.6 h for the wild type (GSTpi(+/+)). Retroviral transfection of GSTP1-1 into GSTpi(-/-) MEF cells slowed the doubling time to 30.4 h. Both early passage and immortalized MEF cells from GSTpi(-/-) animals expressed significantly elevated activity of extracellular signal-regulated kinases ERK1/ERK2, kinases linked to cell proliferation pathways. In vivo, GSTpi(-/-) mice had higher basal levels of circulating white blood cells compared with GSTpi(+/+). Administration of a peptidomimetic inhibitor of GSTP1-1, TLK199, (gamma-glutamyl-S-(benzyl)cysteinyl R-phenyl glycine diethyl ester), stimulated both lymphocyte production and bone marrow progenitor (colony-forming unit-granulocyte macrophage) proliferation, but only in GSTpi(+/+) and not in GSTpi(-/-) animals. Selection of a resistant clone of an HL60 tumor cell line through chronic exposure to TLK199 resulted in cells with elevated activities of c-Jun NH2 terminal kinase (JNK1) and ERK1/ERK2, and allowed the cells to proliferate under stress conditions that induced high levels of apoptosis in the wild type cells. The in vitro and in vivo data are consistent with the principle that GSTP1-1 influences cell proliferation. PMID- 11408561 TI - Stereoselective pharmacokinetics and pharmacodynamics of organic nitrates in rats. AB - Plasma concentration and vasodilating effect after i.v. bolus injection of stereoisomeric organic nitrates were evaluated. Pharmacokinetics of mononitrates was analyzed with a linear one-compartment model. The apparent volumes of distribution were almost identical, but systemic clearances were different among stereoisomers. The concentration data after dinitrate administration could be described based on a two-compartment model with elimination only from the central compartment via metabolism to mononitrate, and then mononitrate-dependent metabolic clearance was estimated. In the vasodilation by mononitrate administered intravenously, the maximum effect was not observed. The reduction of mean arterial pressure from baseline level was related to plasma concentration with a log-linear model. The pharmacological effect following dinitrate dosing was analyzed by a sigmoidal Emax model assuming a simple additive effect of dinitrate and mononitrate. Although almost the same Hill's constant and maximum effect (Emax) values were estimated, the concentrations required to produce 50% of Emax (EC50) differed among stereoisomers. The clearance and EC50 values of stereoisomers with nitrate group at the exo position were generally higher than those with the same group at the endo position. This suggests that the stereostructure of organic nitrates controls the vasodilator potency and duration of action. PMID- 11408562 TI - Effects of T-588, a cognitive enhancer compound, on synaptic plasticity in the dentate gyrus of freely moving rats. AB - (1R)-1-benzo [b] thiophen-5-yl-2-[2-(diethylamino) ethoxy] ethan-1-ol hydrochloride (T-588) is a compound for the treatment of neurodegenerative disorders, including Alzheimer's disease and cerebrovascular diseases. T-588 reportedly alleviates learning and memory deficits in animal models of dementia. In the present study, we investigated the effects of T-588 on the induction and decay of long-term potentiation (LTP) and on the responses to paired-pulse (pp) stimulation in freely moving rats. Perforant path-evoked field potentials were recorded in the dentate gyrus by chronically implanted electrodes. LTP was induced by high-frequency stimulation 30 min after oral administration of T-588 (0.3 or 3 mg/kg). T-588 significantly augmented the increase in population spike amplitude and field excitatory postsynaptic potential slope after LTP induction. T-588 also prolonged the decay of augmented population spike amplitude, but had no significant effect on the response to pp stimulation. These results suggest that T-588 facilitates long-term synaptic plasticity, but not short-term synaptic plasticity in the dentate gyrus of freely moving rats. The effect of T-588 on long-term synaptic plasticity may contribute to the alleviation of learning and memory dysfunction seen in animal models. PMID- 11408563 TI - Early augmentation of interleukin (IL)-12 level in the airway of mice administered orally with clarithromycin or intranasally with IL-12 results in alleviation of influenza infection. AB - The protective role of interleukin (IL)-12 against influenza infection was assessed by analyzing the efficacies of orally administered clarithromycin (CAM) as an immunomodulator and intranasal administration of recombinant IL-12 in intranasally influenza virus-infected mice. In infected mice, CAM at 20 mg/mouse/day significantly elevated the levels of IL-12 and interferon-gamma on days 2 and 3, respectively, after infection in the bronchoalveolar lavage fluid (BALF), but the levels in the sera were not affected. The levels of IL-4, -6, and -10 were not significantly affected in the sera and BALF. Corresponding with the local elevation of IL-12 level, CAM reduced virus yield and the number of infiltrated cells in the BALF, the severity of pneumonia, and mortality of the treated mice. The potential activity of CAM as an experimental immunomodulator was verified at a dose of 20 mg/mouse/day. Intranasal administration of the optimal dose (20 ng/mouse) of IL-12 on day 2 significantly reduced virus yield in the BALF after infection. The loss of body weight was significantly suppressed by IL-12 administration. The local elevation of IL-12 level at the optimal dose and timing in influenza infection was confirmed to be effective in alleviating the influenza infection in mice treated with the two different ways. Thus, the augmentation of IL-12 production or administration of supplementary IL-12 in the respiratory tract was essential in reducing virus yield in the early phase of influenza and may be crucial for recovery from influenza infection. PMID- 11408564 TI - In vivo synergistic interaction of liposome-coencapsulated gentamicin and ceftazidime. AB - Antimicrobial agents may interact synergistically. But to ensure synergy in vivo, the drugs should both be present at the site of infection at sufficiently high concentrations for an adequate period of time. Coencapsulation of the drugs in a drug carrier may ensure parallel tissue distributions. Since liposomes localize preferentially at sites of infection, this mode of drug delivery could, in addition, increase drug concentrations at the focus of infection. The therapeutic efficacy of gentamicin and ceftazidime coencapsulated into liposomes was examined by monitoring survival in a rat model of an acute unilateral pneumonia caused by antibiotic-susceptible and antibiotic-resistant Klebsiella pneumoniae strains. It is shown that administration of gentamicin in combination with ceftazidime in the free form either as single dose or as 5-day treatment resulted in an additive effect on rat survival in both models. In contrast, targeted delivery of liposome coencapsulated gentamicin and ceftazidime resulted in a synergistic interaction of the antibiotics in both models. Consequently, liposome coencapsulation of gentamicin and ceftazidime allowed both a shorter course of treatment at lower cumulative doses compared with administration of the antibiotics in the free form to obtain complete survival of rats. Liposomal coencapsulation of synergistic antibiotics may open new perspectives in the treatment of severe infections. PMID- 11408565 TI - BMS-229724 is a tight-binding inhibitor of cytosolic phospholipase A2 that acts at the lipid/water interface and possesses anti-inflammatory activity in skin inflammation models. AB - Cytosolic phospholipase A2 (cPLA2) catalyzes the selective release of arachidonic acid from the sn-2 position of phospholipids and is believed to play a key cellular role in the generation of arachidonic acid. BMS-229724 (4-[4-[2-[2 [bis(4-chlorophenyl)methoxy]ethyl-sulfonyl]ethoxy]phenyl]-1,1,1-trifluoro-2 butanone) was found to be a selective inhibitor of cPLA2 (IC50 = 2.8 microM) in that it did not inhibit secreted phospholipase A2 in vitro, nor phospholipase C and phospholipase D in cells. The compound was active in inhibiting arachidonate and eicosanoid production in U937 cells, neutrophils, platelets, monocytes, and mast cells. With a synthetic covesicle substrate system, the dose-dependent inhibition could be defined by kinetic equations describing competitive inhibition at the lipid/water interface. The apparent equilibrium dissociation constant for the inhibitor bound to the enzyme at the interface (K(I)*(app)) was determined to be 1. 10(-5) mol% versus an apparent dissociation constant for the arachidonate-containing phospholipid of 0.35 mol%. The unit of concentration in the interface is mole fraction (or mol%), which is related to the surface concentration of substrate, rather than bulk concentration that has units of molarity. Thus, BMS-229724 represents a novel inhibitor of cPLA2, which partitions into the phospholipid bilayer and competes with phospholipid substrate for the active site. This potent inhibition of the enzyme translated into anti inflammatory activity when applied topically (5%, w/v) to a phorbol ester-induced chronic inflammation model in mouse ears, inhibiting edema and neutrophil infiltration, as well as prostaglandin and leukotriene levels in the skin. In hairless guinea pigs, BMS-229724 was active orally (10 mg/kg) in a UVB-induced skin erythema model in hairless guinea pigs. PMID- 11408566 TI - Effects of JL13, a pyridobenzoxazepine with potential atypical antipsychotic activity, in animal models for schizophrenia. AB - JL13 [5-(4-methylpiperazin-1-yl)-8-chloro-pyrido[2,3-b][1,5] benzoxazepine fumarate] is a substance with a close structural resemblance to clozapine. However, it is less sensitive to oxidation and may therefore have less hematological side effects. In the present study, JL13 was compared with clozapine and haloperidol in several animal models for schizophrenia. The paw test represents a screening model for antipsychotic drugs that can discriminate between drugs with extrapyramidal side effects and drugs without. Haloperidol increased both forelimb retraction time and hindlimb retraction time (HRT), whereas both clozapine and JL13 increased only HRT. In the prepulse inhibition paradigm, all three drugs reversed the apomorphine- and the amphetamine-induced disruption of prepulse inhibition. However, whereas haloperidol was equally effective against both dopaminergic drugs, JL13 and clozapine were more effective against amphetamine. Finally, only JL13 was able to increase prepulse inhibition in normal rats, whereas only clozapine reduced basal startle amplitude. Taken together, these data suggest that JL13 may be an effective antipsychotic drug, with a profile similar to clozapine. PMID- 11408567 TI - Ras regulates the polarity of the yeast actin cytoskeleton through the stress response pathway. AB - Polarized growth in yeast requires cooperation between the polarized actin cytoskeleton and delivery of post-Golgi secretory vesicles. We have previously reported that loss of the major tropomyosin isoform, Tpm1p, results in cells sensitive to perturbations in cell polarity. To identify components that bridge these processes, we sought mutations with both a conditional defect in secretion and a partial defect in polarity. Thus, we set up a genetic screen for mutations that conferred a conditional growth defect, showed synthetic lethality with tpm1Delta, and simultaneously became denser at the restrictive temperature, a hallmark of secretion-defective cells. Of the 10 complementation groups recovered, the group with the largest number of independent isolates was functionally null alleles of RAS2. Consistent with this, ras2Delta and tpm1Delta are synthetically lethal at 35 degrees C. We show that ras2Delta confers temperature-sensitive growth and temperature-dependent depolarization of the actin cytoskeleton. Furthermore, we show that at elevated temperatures ras2Delta cells are partially defective in endocytosis and show a delocalization of two key polarity markers, Myo2p and Cdc42p. However, the conditional enhanced density phenotype of ras2Delta cells is not a defect in secretion. All the phenotypes of ras2Delta cells can be fully suppressed by expression of yeast RAS1 or RAS2 genes, human Ha-ras, or the double disruption of the stress response genes msn2Deltamsn4Delta. Although the best characterized pathway of Ras function in yeast involves activation of the cAMP-dependent protein kinase A pathway, activation of the protein kinase A pathway does not fully suppress the actin polarity defects, suggesting that there is an additional pathway from Ras2p to Msn2/4p. Thus, Ras2p regulates cytoskeletal polarity in yeast under conditions of mild temperature stress through the stress response pathway. PMID- 11408568 TI - Formation of a normal epidermis supported by increased stability of keratins 5 and 14 in keratin 10 null mice. AB - The expression of distinct keratin pairs during epidermal differentiation is assumed to fulfill specific and essential cytoskeletal functions. This is supported by a great variety of genodermatoses exhibiting tissue fragility because of keratin mutations. Here, we show that the loss of K10, the most prominent epidermal protein, allowed the formation of a normal epidermis in neonatal mice without signs of fragility or wound-healing response. However, there were profound changes in the composition of suprabasal keratin filaments. K5/14 persisted suprabasally at elevated protein levels, whereas their mRNAs remained restricted to the basal keratinocytes. This indicated a novel mechanism regulating keratin turnover. Moreover, the amount of K1 was reduced. In the absence of its natural partner we observed the formation of a minor amount of novel K1/14/15 filaments as revealed by immunogold electron microscopy. We suggest that these changes maintained epidermal integrity. Furthermore, suprabasal keratinocytes contained larger keratohyalin granules similar to our previous K10T mice. A comparison of profilaggrin processing in K10T and K10(-/-) mice revealed an accumulation of filaggrin precursors in the former but not in the latter, suggesting a requirement of intact keratin filaments for the processing. The mild phenotype of K10(-/-) mice suggests that there is a considerable redundancy in the keratin gene family. PMID- 11408569 TI - Involvement of p38 in apoptosis-associated membrane blebbing and nuclear condensation. AB - The stress-activated protein kinase p38 is often induced by cytotoxic agents, but its contribution to cell death is ill defined. In Rat-1 cells, we found a strong correlation between activation of p38 and induction of c-Myc-dependent apoptosis. In cells with deregulated c-Myc expression but not in control cells, cis diamminedichloroplatinum induced p38 activity and typical features of apoptosis, including internucleosomal DNA degradation, induction of caspase activities, and both nuclear (nuclear condensation and fragmentation) and extranuclear (cell blebbing) morphological alterations. The pan-caspase inhibitor N benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone did not block p38 activation and the p38 inhibitor SB203580 had no detectable effect on the activation of caspases or the in vivo cleavage of several caspase substrates, suggesting that p38 and caspase activation can contribute distinct features of apoptosis. Accordingly, we found that cell blebbing was independent of caspase activity and, rather, depended on p38-sensitive changes in microfilament dynamics likely mediated by heat shock protein 27 phosphorylation. Furthermore, p38 activity contributed to both caspase-dependent and caspase-independent nuclear condensation and fragmentation, suggesting a role in an early event triggering both mechanisms of apoptosis or sensitizing the cells to the action of both types of apoptosis executioners. Inhibiting p38 also resulted in a significant enhancement in cell survival estimated by colony formation. This capacity to modulate the sensitivity to apoptosis in cells with deregulated c-Myc expression suggests an important role for p38 in tumor cell killing by chemotherapeutic agents. PMID- 11408570 TI - Characterization of a novel human SMC heterodimer homologous to the Schizosaccharomyces pombe Rad18/Spr18 complex. AB - The structural maintenance of chromosomes (SMC) protein encoded by the fission yeast rad18 gene is involved in several DNA repair processes and has an essential function in DNA replication and mitotic control. It has a heterodimeric partner SMC protein, Spr18, with which it forms the core of a multiprotein complex. We have now isolated the human orthologues of rad18 and spr18 and designated them hSMC6 and hSMC5. Both proteins are about 1100 amino acids in length and are 27 28% identical to their fission yeast orthologues, with much greater identity within their N- and C-terminal globular domains. The hSMC6 and hSMC5 proteins interact to form a tight complex analogous to the yeast Rad18/Spr18 heterodimer. In proliferating human cells the proteins are bound to both chromatin and the nucleoskeleton. In addition, we have detected a phosphorylated form of hSMC6 that localizes to interchromatin granule clusters. Both the total level of hSMC6 and its phosphorylated form remain constant through the cell cycle. Both hSMC5 and hSMC6 proteins are expressed at extremely high levels in the testis and associate with the sex chromosomes in the late stages of meiotic prophase, suggesting a possible role for these proteins in meiosis. PMID- 11408571 TI - alpha-catenin-independent recruitment of ZO-1 to nectin-based cell-cell adhesion sites through afadin. AB - ZO-1 is an actin filament (F-actin)-binding protein that localizes to tight junctions and connects claudin to the actin cytoskeleton in epithelial cells. In nonepithelial cells that have no tight junctions, ZO-1 localizes to adherens junctions (AJs) and may connect cadherin to the actin cytoskeleton indirectly through beta- and alpha-catenins as one of many F-actin-binding proteins. Nectin is an immunoglobulin-like adhesion molecule that localizes to AJs and is associated with the actin cytoskeleton through afadin, an F-actin-binding protein. Ponsin is an afadin- and vinculin-binding protein that also localizes to AJs. The nectin-afadin complex has a potency to recruit the E-cadherin-beta catenin complex through alpha-catenin in a manner independent of ponsin. By the use of cadherin-deficient L cell lines stably expressing various components of the cadherin-catenin and nectin-afadin systems, and alpha-catenin-deficient F9 cell lines, we examined here whether nectin recruits ZO-1 to nectin-based cell cell adhesion sites. Nectin showed a potency to recruit not only alpha-catenin but also ZO-1 to nectin-based cell-cell adhesion sites. This recruitment of ZO-1 was dependent on afadin but independent of alpha-catenin and ponsin. These results indicate that ZO-1 localizes to cadherin-based AJs through interactions not only with alpha-catenin but also with the nectin-afadin system. PMID- 11408572 TI - SPO21 is required for meiosis-specific modification of the spindle pole body in yeast. AB - During meiosis II in the yeast Saccharomyces cerevisiae, the cytoplasmic face of the spindle pole body changes from a site of microtubule initiation to a site of de novo membrane formation. These membranes are required to package the haploid meiotic products into spores. This functional change in the spindle pole body involves the expansion and modification of its cytoplasmic face, termed the outer plaque. We report here that SPO21 is required for this modification. The Spo21 protein localizes to the spindle pole in meiotic cells. In the absence of SPO21 the structure of the outer plaque is abnormal, and prospore membranes do not form. Further, decreased dosage of SPO21 leaves only two of the four spindle pole bodies competent to generate membranes. Mutation of CNM67, encoding a known component of the mitotic outer plaque, also results in a meiotic outer plaque defect but does not block membrane formation, suggesting that Spo21p may play a direct role in initiating membrane formation. PMID- 11408573 TI - Lysosomal hydrolase mannose 6-phosphate uncovering enzyme resides in the trans Golgi network. AB - A crucial step in lysosomal biogenesis is catalyzed by "uncovering" enzyme (UCE), which removes a covering N-acetylglucosamine from the mannose 6-phosphate (Man-6 P) recognition marker on lysosomal hydrolases. This study shows that UCE resides in the trans-Golgi network (TGN) and cycles between the TGN and plasma membrane. The cytosolic domain of UCE contains two potential endocytosis motifs: (488)YHPL and C-terminal (511)NPFKD. YHPL is shown to be the more potent of the two in retrieval of UCE from the plasma membrane. A green-fluorescent protein-UCE transmembrane-cytosolic domain fusion protein colocalizes with TGN 46, as does endogenous UCE in HeLa cells, showing that the transmembrane and cytosolic domains determine intracellular location. These data imply that the Man-6-P recognition marker is formed in the TGN, the compartment where Man-6-P receptors bind cargo and are packaged into clathrin-coated vesicles. PMID- 11408574 TI - Dual targeting of Osh1p, a yeast homologue of oxysterol-binding protein, to both the Golgi and the nucleus-vacuole junction. AB - Oxysterol binding protein (OSBP) is the only protein known to bind specifically to the group of oxysterols with potent effects on cholesterol homeostasis. Although the function of OSBP is currently unknown, an important role is implicated by the existence of multiple homologues in all eukaryotes so far examined. OSBP and a subset of homologues contain pleckstrin homology (PH) domains. Such domains are responsible for the targeting of a wide range of proteins to the plasma membrane. In contrast, OSBP is a peripheral protein of Golgi membranes, and its PH domain targets to the trans-Golgi network of mammalian cells. In this article, we have characterized Osh1p, Osh2p, and Osh3p, the three homologues of OSBP in Saccharomyces cerevisiae that contain PH domains. Examination of a green fluorescent protein (GFP) fusion to Osh1p revealed a striking dual localization with the protein present on both the late Golgi, and in the recently described nucleus-vacuole (NV) junction. Deletion mapping revealed that the PH domain of Osh1p specified targeting to the late Golgi, and an ankyrin repeat domain targeting to the NV junction, the first such targeting domain identified for this structure. GFP fusions to Osh2p and Osh3p showed intracellular distributions distinct from that of Osh1p, and their PH domains appear to contribute to their differing localizations. PMID- 11408575 TI - Dynamic localization of the Swe1 regulator Hsl7 during the Saccharomyces cerevisiae cell cycle. AB - In Saccharomyces cerevisiae, entry into mitosis requires activation of the cyclin dependent kinase Cdc28 in its cyclin B (Clb)-associated form. Clb-bound Cdc28 is susceptible to inhibitory tyrosine phosphorylation by Swe1 protein kinase. Swe1 is itself negatively regulated by Hsl1, a Nim1-related protein kinase, and by Hsl7, a presumptive protein-arginine methyltransferase. In vivo all three proteins localize to the bud neck in a septin-dependent manner, consistent with our previous proposal that formation of Hsl1-Hsl7-Swe1 complexes constitutes a checkpoint that monitors septin assembly. We show here that Hsl7 is phosphorylated by Hsl1 in immune-complex kinase assays and can physically associate in vitro with either Hsl1 or Swe1 in the absence of any other yeast proteins. With the use of both the two-hybrid method and in vitro binding assays, we found that Hsl7 contains distinct binding sites for Hsl1 and Swe1. A differential interaction trap approach was used to isolate four single-site substitution mutations in Hsl7, which cluster within a discrete region of its N terminal domain, that are specifically defective in binding Hsl1. When expressed in hsl7Delta cells, each of these Hsl7 point mutants is unable to localize at the bud neck and cannot mediate down-regulation of Swe1, but retains other functions of Hsl7, including oligomerization and association with Swe1. GFP-fusions of these Hsl1-binding defective Hsl7 proteins localize as a bright perinuclear dot, but never localize to the bud neck; likewise, in hsl1Delta cells, a GFP-fusion to wild-type Hsl7 or native Hsl7 localizes to this dot. Cell synchronization studies showed that, normally, Hsl7 localizes to the dot, but only in cells in the G1 phase of the cell cycle. Immunofluorescence analysis and immunoelectron microscopy established that the dot corresponds to the outer plaque of the spindle pole body (SPB). These data demonstrate that association between Hsl1 and Hsl7 at the bud neck is required to alleviate Swe1-imposed G2-M delay. Hsl7 localization at the SPB during G1 may play some additional role in fine-tuning the coordination between nuclear and cortical events before mitosis. PMID- 11408576 TI - Drosophila heterochromatin protein 1 (HP1)/origin recognition complex (ORC) protein is associated with HP1 and ORC and functions in heterochromatin-induced silencing. AB - Heterochromatin protein 1 (HP1) is a conserved component of the highly compact chromatin of higher eukaryotic centromeres and telomeres. Cytogenetic experiments in Drosophila have shown that HP1 localization into this chromatin is perturbed in mutants for the origin recognition complex (ORC) 2 subunit. ORC has a multisubunit DNA-binding activity that binds origins of DNA replication where it is required for origin firing. The DNA-binding activity of ORC is also used in the recruitment of the Sir1 protein to silence nucleation sites flanking silent copies of the mating-type genes in Saccharomyces cerevisiae. A fraction of HP1 in the maternally loaded cytoplasm of the early Drosophila embryo is associated with a multiprotein complex containing Drosophila melanogaster ORC subunits. This complex appears to be poised to function in heterochromatin assembly later in embryonic development. Here we report the identification of a novel component of this complex, the HP1/ORC-associated protein. This protein contains similarity to DNA sequence-specific HMG proteins and is shown to bind specific satellite sequences and the telomere-associated sequence in vitro. The protein is shown to have heterochromatic localization in both diploid interphase and mitotic chromosomes and polytene chromosomes. Moreover, the gene encoding HP1/ORC associated protein was found to display reciprocal dose-dependent variegation modifier phenotypes, similar to those for mutants in HP1 and the ORC 2 subunit. PMID- 11408577 TI - Outer dense fiber 2 is a widespread centrosome scaffold component preferentially associated with mother centrioles: its identification from isolated centrosomes. AB - Because centrosomes were enriched in the bile canaliculi fraction from the chicken liver through their association with apical membranes, we developed a procedure for isolation of centrosomes from this fraction. With the use of the centrosomes, we generated centrosome-specific monoclonal antibodies. Three of the monoclonal antibodies recognized an antigen of ~90 kDa. Cloning of its cDNA identified this antigen as a chicken homologue of outer dense fiber 2 protein (Odf2), which was initially identified as a sperm outer dense fiber-specific component. Exogenously expressed and endogenous Odf2 were shown to be concentrated at the centrosomes in a microtubule-independent manner in various types of cells at both light and electron microscopic levels. Odf2 exhibited a cell cycle-dependent pattern of localization and was preferentially associated with the mother centrioles in G0/G1-phase. Toward G1/S-phase before centrosome duplication, it became detectable in both mother and daughter centrioles. In the isolated bile canaliculi and centrosomes, Odf2, in contrast to other centrosomal components, was highly resistant to KI extraction. These findings indicate that Odf2 is a widespread KI-insoluble scaffold component of the centrosome matrix, which may be involved in the maturation event of daughter centrioles. PMID- 11408578 TI - Homotypic fusion of immature secretory granules during maturation requires syntaxin 6. AB - Homotypic fusion of immature secretory granules (ISGs) gives rise to mature secretory granules (MSGs), the storage compartment in endocrine and neuroendocrine cells for hormones and neuropeptides. With the use of a cell-free fusion assay, we investigated which soluble N-ethylmaleimide-sensitive fusion protein attachment receptor (SNARE) molecules are involved in the homotypic fusion of ISGs. Interestingly, the SNARE molecules mediating the exocytosis of MSGs in neuroendocrine cells, syntaxin 1, SNAP-25, and VAMP2, were not involved in homotypic ISG fusion. Instead, we have identified syntaxin 6 as a component of the core machinery responsible for homotypic ISG fusion. Subcellular fractionation studies and indirect immunofluorescence microscopy show that syntaxin 6 is sorted away during the maturation of ISGs to MSGs. Although, syntaxin 6 on ISG membranes is associated with SNAP-25 and SNAP-29/GS32, we could not find evidence that these target (t)-SNARE molecules are involved in homotypic ISG fusion. Nor could we find any involvement for the vesicle (v)-SNARE VAMP4, which is known to be associated with syntaxin 6. Importantly, we have shown that homotypic fusion requires the function of syntaxin 6 on both donor as well as acceptor membranes, which suggests that t-t-SNARE interactions, either direct or indirect, may be required during fusion of ISG membranes. PMID- 11408579 TI - A novel quality control compartment derived from the endoplasmic reticulum. AB - Degradation of proteins that, because of improper or suboptimal processing, are retained in the endoplasmic reticulum (ER) involves retrotranslocation to reach the cytosolic ubiquitin-proteasome machinery. We found that substrates of this pathway, the precursor of human asialoglycoprotein receptor H2a and free heavy chains of murine class I major histocompatibility complex (MHC), accumulate in a novel preGolgi compartment that is adjacent to but not overlapping with the centrosome, the Golgi complex, and the ER-to-Golgi intermediate compartment (ERGIC). On its way to degradation, H2a associated increasingly after synthesis with the ER translocon Sec61. Nevertheless, it remained in the secretory pathway upon proteasomal inhibition, suggesting that its retrotranslocation must be tightly coupled to the degradation process. In the presence of proteasomal inhibitors, the ER chaperones calreticulin and calnexin, but not BiP, PDI, or glycoprotein glucosyltransferase, concentrate in the subcellular region of the novel compartment. The "quality control" compartment is possibly a subcompartment of the ER. It depends on microtubules but is insensitive to brefeldin A. We discuss the possibility that it is also the site for concentration and retrotranslocation of proteins that, like the mutant cystic fibrosis transmembrane conductance regulator, are transported to the cytosol, where they form large aggregates, the "aggresomes." PMID- 11408580 TI - Distribution and transport of cholesterol in Caenorhabditis elegans. AB - Cholesterol transport is an essential process in all multicellular organisms. In this study we applied two recently developed approaches to investigate the distribution and molecular mechanisms of cholesterol transport in Caenorhabditis elegans. The distribution of cholesterol in living worms was studied by imaging its fluorescent analog, dehydroergosterol, which we applied to the animals by feeding. Dehydroergosterol accumulates primarily in the pharynx, nerve ring, excretory gland cell, and gut of L1-L3 larvae. Later, the bulk of dehydroergosterol accumulates in oocytes and spermatozoa. Males display exceptionally strong labeling of spermatids, which suggests a possible role for cholesterol in sperm development. In a complementary approach, we used a photoactivatable cholesterol analog to identify cholesterol-binding proteins in C. elegans. Three major and several minor proteins were found specifically cross linked to photocholesterol after UV irradiation. The major proteins were identified as vitellogenins. rme-2 mutants, which lack the vitellogenin receptor, fail to accumulate dehydroergosterol in oocytes and embryos and instead accumulate dehydroergosterol in the body cavity along with vitellogenin. Thus, uptake of cholesterol by C. elegans oocytes occurs via an endocytotic pathway involving yolk proteins. The pathway is a likely evolutionary ancestor of mammalian cholesterol transport. PMID- 11408581 TI - The human cytomegalovirus US28 protein is located in endocytic vesicles and undergoes constitutive endocytosis and recycling. AB - Genes encoding chemokine receptor-like proteins have been found in herpes and poxviruses and implicated in viral pathogenesis. Here we describe the cellular distribution and trafficking of a human cytomegalovirus (HCMV) chemokine receptor encoded by the US28 gene, after transient and stable expression in transfected HeLa and Cos cells. Immunofluorescence staining indicated that this viral protein accumulated intracellularly in vesicular structures in the perinuclear region of the cell and showed overlap with markers for endocytic organelles. By immunogold electron microscopy US28 was seen mostly to localize to multivesicular endosomes. A minor portion of the protein (at most 20%) was also expressed at the cell surface. Antibody-feeding experiments indicated that cell surface US28 undergoes constitutive ligand-independent endocytosis. Biochemical analysis with the use of iodinated ligands showed that US28 was rapidly internalized. The high-affinity ligand of US28, the CX(3)C-chemokine fractalkine, reduced the steady-state levels of US28 at the cell surface, apparently by inhibiting the recycling of internalized receptor. Endocytosis and cycling of HCMV US28 could play a role in the sequestration of host chemokines, thereby modulating antiviral immune responses. In addition, the distribution of US28 mainly on endosomal membranes may allow it to be incorporated into the viral envelope during HCMV assembly. PMID- 11408582 TI - Spindle dynamics and the role of gamma-tubulin in early Caenorhabditis elegans embryos. AB - gamma-Tubulin is a ubiquitous and highly conserved component of centrosomes in eukaryotic cells. Genetic and biochemical studies have demonstrated that gamma tubulin functions as part of a complex to nucleate microtubule polymerization from centrosomes. We show that, as in other organisms, Caenorhabditis elegans gamma-tubulin is concentrated in centrosomes. To study centrosome dynamics in embryos, we generated transgenic worms that express GFP::gamma-tubulin or GFP::beta-tubulin in the maternal germ line and early embryos. Multiphoton microscopy of embryos produced by these worms revealed the time course of daughter centrosome appearance and growth and the differential behavior of centrosomes destined for germ line and somatic blastomeres. To study the role of gamma-tubulin in nucleation and organization of spindle microtubules, we used RNA interference (RNAi) to deplete C. elegans embryos of gamma-tubulin. gamma-Tubulin (RNAi) embryos failed in chromosome segregation, but surprisingly, they contained extensive microtubule arrays. Moderately affected embryos contained bipolar spindles with dense and long astral microtubule arrays but with poorly organized kinetochore and interpolar microtubules. Severely affected embryos contained collapsed spindles with numerous long astral microtubules. Our results suggest that gamma-tubulin is not absolutely required for microtubule nucleation in C. elegans but is required for the normal organization and function of kinetochore and interpolar microtubules. PMID- 11408583 TI - Cell adhesion molecule L1 in folded (horseshoe) and extended conformations. AB - We have investigated the structure of the cell adhesion molecule L1 by electron microscopy. We were particularly interested in the conformation of the four N terminal immunoglobulin domains, because x-ray diffraction showed that these domains are bent into a horseshoe shape in the related molecules hemolin and axonin-1. Surprisingly, rotary-shadowed specimens showed the molecules to be elongated, with no indication of the horseshoe shape. However, sedimentation data suggested that these domains of L1 were folded into a compact shape in solution; therefore, this prompted us to look at the molecules by an alternative technique, negative stain. The negative stain images showed a compact shape consistent with the expected horseshoe conformation. We speculate that in rotary shadowing the contact with the mica caused a distortion of the protein, weakening the bonds forming the horseshoe and permitting the molecule to extend. We have thus confirmed that the L1 molecule is primarily in the horseshoe conformation in solution, and we have visualized for the first time its opening into an extended conformation. Our study resolves conflicting interpretations from previous electron microscopy studies of L1. PMID- 11408584 TI - Complete cytolysis and neonatal lethality in keratin 5 knockout mice reveal its fundamental role in skin integrity and in epidermolysis bullosa simplex. AB - In human patients, a wide range of mutations in keratin (K) 5 or K14 lead to the blistering skin disorder epidermolysis bullosa simplex. Given that K14 deficiency does not lead to the ablation of a basal cell cytoskeleton because of a compensatory role of K15, we have investigated the requirement for the keratin cytoskeleton in basal cells by inactivating the K5 gene in mice. We report that the K5(-/-) mice die shortly after birth, lack keratin filaments in the basal epidermis, and are more severely affected than K14(-/-) mice. In contrast to the K14(-/-) mice, we detected a strong induction of the wound-healing keratin K6 in the suprabasal epidermis of cytolyzed areas of postnatal K5(-/-) mice. In addition, K5 and K14 mice differed with respect to tongue lesions. Moreover, we show that in the absence of K5 and other type II keratins, residual K14 and K15 aggregated along hemidesmosomes, demonstrating that individual keratins without a partner are stable in vivo. Our data indicate that K5 may be the natural partner of K15 and K17. We suggest that K5 null mutations may be lethal in human epidermolysis bullosa simplex patients. PMID- 11408585 TI - The polo-like kinase Plx1 is required for activation of the phosphatase Cdc25C and cyclin B-Cdc2 in Xenopus oocytes. AB - In the Xenopus oocyte system mitogen treatment triggers the G(2)/M transition by transiently inhibiting the cAMP-dependent protein kinase (PKA); subsequently, other signal transduction pathways are activated, including the mitogen-activated protein kinase (MAPK) and polo-like kinase pathways. To study the interactions between these pathways, we have utilized a cell-free oocyte extract that carries out the signaling events of oocyte maturation after addition of the heat-stable inhibitor of PKA, PKI. PKI stimulated the synthesis of Mos and activation of both the MAPK pathway and the Plx1/Cdc25C/cyclin B-Cdc2 pathway. Activation of the MAPK pathway alone by glutathione S-transferase (GST)-Mos did not lead to activation of Plx1 or cyclin B-Cdc2. Inhibition of the MAPK pathway in the extract by the MEK1 inhibitor U0126 delayed, but did not prevent, activation of the Plx1 pathway, and inhibition of Mos synthesis by cycloheximide had a similar effect, suggesting that MAPK activation is the only relevant function of Mos. Immunodepletion of Plx1 completely inhibited activation of Cdc25C and cyclin B Cdc2 by PKI, indicating that Plx1 is necessary for Cdc25C activation. In extracts containing fully activated Plx1 and Cdc25C, inhibition of cyclin B-Cdc2 by p21(Cip1) had no significant effect on either the phosphorylation of Cdc25C or the activity of Plx1. These results demonstrate that maintenance of Plx1 and Cdc25C activity during mitosis does not require cyclin B-Cdc2 activity. PMID- 11408586 TI - Identification of a Saccharomyces cerevisiae gene that is required for G1 arrest in response to the lipid oxidation product linoleic acid hydroperoxide. AB - Reactive oxygen species cause damage to all of the major cellular constituents, including peroxidation of lipids. Previous studies have revealed that oxidative stress, including exposure to oxidation products, affects the progression of cells through the cell division cycle. This study examined the effect of linoleic acid hydroperoxide, a lipid peroxidation product, on the yeast cell cycle. Treatment with this peroxide led to accumulation of unbudded cells in asynchronous populations, together with a budding and replication delay in synchronous ones. This observed modulation of G1 progression could be distinguished from the lethal effects of the treatment and may have been due to a checkpoint mechanism, analogous to that known to be involved in effecting cell cycle arrest in response to DNA damage. By examining several mutants sensitive to linoleic acid hydroperoxide, the YNL099c open reading frame was found to be required for the arrest. This gene (designated OCA1) encodes a putative protein tyrosine phosphatase of previously unknown function. Cells lacking OCA1 did not accumulate in G1 on treatment with linoleic acid hydroperoxide, nor did they show a budding, replication, or Start delay in synchronous cultures. Although not essential for adaptation or immediate cellular survival, OCA1 was required for growth in the presence of linoleic acid hydroperoxide, thus indicating that it may function in linking growth, stress responses, and the cell cycle. Identification of OCA1 establishes cell cycle arrest as an actively regulated response to oxidative stress and will enable further elucidation of oxidative stress-responsive signaling pathways in yeast. PMID- 11408587 TI - Mitotic phosphorylation of Golgi reassembly stacking protein 55 by mitogen activated protein kinase ERK2. AB - The role of the mitogen-activated protein kinase kinase (MKK)/extracellular activated protein kinase (ERK) pathway in mitotic Golgi disassembly is controversial, in part because Golgi-localized targets have not been identified. We observed that Golgi reassembly stacking protein 55 (GRASP55) was phosphorylated in mitotic cells and extracts, generating a mitosis-specific phospho-epitope recognized by the MPM2 mAb. This phosphorylation was prevented by mutation of ERK consensus sites in GRASP55. GRASP55 mitotic phosphorylation was significantly reduced, both in vitro and in vivo, by treatment with U0126, a potent and specific inhibitor of MKK and thus ERK activation. Furthermore, ERK2 directly phosphorylated GRASP55 on the same residues that generated the MPM2 phospho-epitope. These results are the first demonstration of GRASP55 mitotic phosphorylation and indicate that the MKK/ERK pathway directly phosphorylates the Golgi during mitosis. PMID- 11408588 TI - Sphingomyelin-enriched microdomains at the Golgi complex. AB - Sphingomyelin- and cholesterol-enriched microdomains can be isolated as detergent resistant membranes from total cell extracts (total-DRM). It is generally believed that this total-DRM represents microdomains of the plasma membrane. Here we describe the purification and detailed characterization of microdomains from Golgi membranes. These Golgi-derived detergent-insoluble complexes (GICs) have a low buoyant density and are highly enriched in lipids, containing 25% of total Golgi phospholipids including 67% of Golgi-derived sphingomyelin, and 43% of Golgi-derived cholesterol. In contrast to total-DRM, GICs contain only 10 major proteins, present in nearly stoichiometric amounts, including the alpha- and beta subunits of heterotrimeric G proteins, flotillin-1, caveolin, and subunits of the vacuolar ATPase. Morphological data show a brefeldin A-sensitive and temperature sensitive localization to the Golgi complex. Strikingly, the stability of GICs does not depend on its membrane environment, because, after addition of brefeldin A to cells, GICs can be isolated from a fused Golgi-endoplasmic reticulum organelle. This indicates that GIC microdomains are not in a dynamic equilibrium with neighboring membrane proteins and lipids. After disruption of the microdomains by cholesterol extraction with cyclodextrin, a subcomplex of several GIC proteins including the B-subunit of the vacuolar ATPase, flotillin-1, caveolin, and p17 could still be isolated by immunoprecipitation. This indicates that several of the identified GIC proteins localize to the same microdomains and that the microdomain scaffold is not required for protein interactions between these GIC proteins but instead might modulate their affinity. PMID- 11408589 TI - Perturbation of the nucleus: a novel Hog1p-independent, Pkc1p-dependent consequence of hypertonic shock in yeast. AB - Hypertonic shock of Saccharomyces cerevisiae activates the Hog1p MAP kinase cascade. In contrast, protein kinase C (Pkc1p) and the "cell integrity" MAP kinase cascade are critical for the response to hypotonic shock. We observed that hypertonic shock transiently relocated many, but not all, nuclear and nucleolar proteins to the cytoplasm. We hypothesized that the relocation of nuclear proteins was due to activation of the Hog1p kinase cascade, yet, surprisingly, Hog1p was not required for these effects. In contrast, Pkc1p kinase activity was required, although the Pkc1p MAP kinase cascade and several factors known to lie upstream and downstream of Pkc1p were not. Moreover, sudden induction of a hyperactive form of Pkc1p was sufficient to relocate nuclear proteins. Taken together, these observations show that the scope of involvement of Pkc1p in the organization of the nucleus considerably exceeds what has been characterized previously. The relocation of nuclear proteins is likely to account for the profound inhibition of RNA synthesis that was observed during hypertonic shock. PMID- 11408590 TI - Myosin vb is associated with plasma membrane recycling systems. AB - Myosin Va is associated with discrete vesicle populations in a number of cell types, but little is known of the function of myosin Vb. Yeast two-hybrid screening of a rabbit parietal cell cDNA library with dominant active Rab11a (Rab11aS20V) identified myosin Vb as an interacting protein for Rab11a, a marker for plasma membrane recycling systems. The isolated clone, corresponding to the carboxyl terminal 60 kDa of the myosin Vb tail, interacted with all members of the Rab11 family (Rab11a, Rab11b, and Rab25). GFP-myosin Vb and endogenous myosin Vb immunoreactivity codistributed with Rab11a in HeLa and Madin-Darby canine kidney (MDCK) cells. As with Rab11a in MDCK cells, the myosin Vb immunoreactivity was dispersed with nocodazole treatment and relocated to the apical corners of cells with taxol treatment. A green fluorescent protein (GFP)-myosin Vb tail chimera overexpressed in HeLa cells retarded transferrin recycling and caused accumulation of transferrin and the transferrin receptor in pericentrosomal vesicles. Expression of the myosin Vb tail chimera in polarized MDCK cells stably expressing the polymeric IgA receptor caused accumulation of basolaterally endocytosed polymeric IgA and the polymeric IgA receptor in the pericentrosomal region. The myosin Vb tail had no effects on transferrin trafficking in polarized MDCK cells. The GFP-myosin Va tail did not colocalize with Rab11a and had no effects on recycling system vesicle distribution in either HeLa or MDCK cells. The results indicate myosin Vb is associated with the plasma membrane recycling system in nonpolarized cells and the apical recycling system in polarized cells. The dominant negative effects of the myosin Vb tail chimera indicate that this unconventional myosin is required for transit out of plasma membrane recycling systems. PMID- 11408591 TI - Hyaluronan activates cell motility of v-Src-transformed cells via Ras-mitogen activated protein kinase and phosphoinositide 3-kinase-Akt in a tumor-specific manner. AB - We investigated the production of hyaluronan (HA) and its effect on cell motility in cells expressing the v-src mutants. Transformation of 3Y1 by v-src virtually activated HA secretion, whereas G2A v-src, a nonmyristoylated form of v-src defective in cell transformation, had no effect. In cells expressing the temperature-sensitive mutant of v-Src, HA secretion was temperature dependent. In addition, HA as small as 1 nM, on the other side, activated cell motility in a tumor-specific manner. HA treatment strongly activated the motility of v-Src transformed 3Y1, whereas it showed no effect on 3Y1- and 3Y1-expressing G2A v src. HA-dependent cell locomotion was strongly blocked by either expression of dominant-negative Ras or treatment with a Ras farnesyltransferase inhibitor. Similarly, both the MEK1 inhibitor and the kinase inhibitor clearly inhibited HA dependent cell locomotion. In contrast, cells transformed with an active MEK1 did not respond to the HA. Finally, an anti-CD44-neutralizing antibody could block the activation of cell motility by HA as well as the HA-dependent phosphorylation of mitogen-activated protein kinase and Akt. Taken together, these results suggest that simultaneous activation of the Ras-mitogen-activated protein kinase pathway and the phosphoinositide 3-kinase pathway by the HA-CD44 interaction is required for the activation of HA-dependent cell locomotion in v-Src-transformed cells. PMID- 11408592 TI - An intact dilysine-like motif in the carboxyl terminus of MAL is required for normal apical transport of the influenza virus hemagglutinin cargo protein in epithelial Madin-Darby canine kidney cells. AB - The MAL proteolipid, a component of the integral protein sorting machinery, has been demonstrated as being necessary for normal apical transport of the influenza virus hemagglutinin (HA) and the overall apical membrane proteins in Madin-Darby canine kidney (MDCK) cells. The MAL carboxy terminus ends with the sequence Arg Trp-Lys-Ser-Ser (RWKSS), which resembles dilysine-based motifs involved in protein sorting. To investigate whether the RWKSS pentapeptide plays a role in modulating the distribution of MAL and/or its function in apical transport, we have expressed MAL proteins with distinct carboxy terminus in MDCK cells whose apical transport was impaired by depletion of endogenous MAL. Apical transport of HA was restored to normal levels by expression of MAL with an intact but not with modified carboxyl terminal sequences bearing mutations that impair the functioning of dilysine-based sorting signals, although all the MAL proteins analyzed incorporated efficiently into lipid rafts. Ultrastructural analysis indicated that compared with MAL bearing an intact RWKSS sequence, a mutant with lysine -3 substituted by serine showed a twofold increased presence in clathrin coated cytoplasmic structures and a reduced expression on the plasma membrane. These results indicate that the carboxyl-terminal RWKSS sequence modulates the distribution of MAL in clathrin-coated elements and is necessary for HA transport to the apical surface. PMID- 11408593 TI - Yeast Gga coat proteins function with clathrin in Golgi to endosome transport. AB - Gga proteins represent a newly recognized, evolutionarily conserved protein family with homology to the "ear" domain of the clathrin adaptor AP-1 gamma subunit. Yeast cells contain two Gga proteins, Gga1p and Gga2p, that have been proposed to act in transport between the trans-Golgi network and endosomes. Here we provide genetic and physical evidence that yeast Gga proteins function in trans-Golgi network clathrin coats. Deletion of Gga2p (gga2Delta), the major Gga protein, accentuates growth and alpha-factor maturation defects in cells carrying a temperature-sensitive allele of the clathrin heavy chain gene. Cells carrying either gga2Delta or a deletion of the AP-1 beta subunit gene (apl2Delta) alone are phenotypically normal, but cells carrying both gga2Delta and apl2Delta are defective in growth, alpha-factor maturation, and transport of carboxypeptidase S to the vacuole. Disruption of both GGA genes and APL2 results in cells so severely compromised in growth that they form only microcolonies. Gga proteins can bind clathrin in vitro and cofractionate with clathrin-coated vesicles. Our results indicate that yeast Gga proteins play an important role in cargo selective clathrin-mediated protein traffic from the trans-Golgi network to endosomes. PMID- 11408594 TI - Regulation of epidermal growth factor receptor signaling by endocytosis and intracellular trafficking. AB - Ligand activation of the epidermal growth factor receptor (EGFR) leads to its rapid internalization and eventual delivery to lysosomes. This process is thought to be a mechanism to attenuate signaling, but signals could potentially be generated after endocytosis. To directly evaluate EGFR signaling during receptor trafficking, we developed a technique to rapidly and selectively isolate internalized EGFR and associated molecules with the use of reversibly biotinylated anti-EGFR antibodies. In addition, we developed antibodies specific to tyrosine-phosphorylated EGFR. With the use of a combination of fluorescence imaging and affinity precipitation approaches, we evaluated the state of EGFR activation and substrate association during trafficking in epithelial cells. We found that after internalization, EGFR remained active in the early endosomes. However, receptors were inactivated before degradation, apparently due to ligand removal from endosomes. Adapter molecules, such as Shc, were associated with EGFR both at the cell surface and within endosomes. Some molecules, such as Grb2, were primarily found associated with surface EGFR, whereas others, such as Eps8, were found only with intracellular receptors. During the inactivation phase, c-Cbl became EGFR associated, consistent with its postulated role in receptor attenuation. We conclude that the association of the EGFR with different proteins is compartment specific. In addition, ligand loss is the proximal cause of EGFR inactivation. Thus, regulated trafficking could potentially influence the pattern as well as the duration of signal transduction. PMID- 11408595 TI - Structural mimicry in G protein-coupled receptors: implications of the high resolution structure of rhodopsin for structure-function analysis of rhodopsin like receptors. AB - The availability of a high-resolution structure of rhodopsin now allows us to reconsider research attempts to understand structure-function relationships in other G protein-coupled receptors (GPCRs). A comparison of the rhodopsin structure with the results of previous sequence analysis and molecular modeling that incorporated experimental results demonstrates a high degree of success for these methods in predicting the helix ends and protein-protein interface of GPCRs. Moreover, the amino acid residues inferred to form the surface of the binding-site crevice based on our application of the substituted-cysteine accessibility method in the dopamine D(2) receptor are in remarkable agreement with the rhodopsin structure, with the notable exception of some residues in the fourth transmembrane segment. Based on our analysis of the data reviewed, we propose that the overall structures of rhodopsin and of amine receptors are very similar, although we also identified localized regions where the structure of these receptors may diverge. We further propose that several of the highly unusual structural features of rhodopsin are also present in amine GPCRs, despite the absence of amino acids that might have thought to have been critical to the adoption of these features. Thus, different amino acids or alternate microdomains can support similar deviations from regular alpha-helical structure, thereby resulting in similar tertiary structure. Such structural mimicry is a mechanism by which a common ancestor could diverge sufficiently to develop the selectivity necessary to interact with diverse signals, while still maintaining a similar overall fold. Through this process, the core function of signaling activation through a conformational change in the transmembrane segments that alters the conformation of the cytoplasmic surface and subsequent interaction with G proteins is presumably shared by the entire Class A family of receptors, despite their selectivity for a diverse group of ligands. PMID- 11408596 TI - Regulation of opioid receptor function by chronic agonist exposure: constitutive activity and desensitization. PMID- 11408597 TI - Cloning, expression, signaling mechanisms, and membrane targeting of P2Y(11) receptors in Madin Darby canine kidney cells. AB - The P2Y(11) receptor is hypothesized to link to both G(s) and G(q), although this proposition is based on expression and separate assays of G(s) and G(q) function in different cell types [J Biol Chem 1997;272:31969-31973]. We have cloned and characterized a canine P2Y(11)-like (cP2Y(11)) receptor from cultured Madin Darby canine kidney (MDCK-D1) cells. When cP2Y(11) receptors are expressed in canine thymocyte (CF2Th) cells that normally lack functional purinergic responses, ADP beta S stimulates phosphatidylinositol (PI) hydrolysis, Ca(2+) mobilization, and cAMP accumulation. Pharmacologic analysis indicates that the stimulation of cAMP production is direct and not a result of eicosanoid synthesis, activation of PKC, or elevation of cell Ca(2+). The rank order of potency for stimulation of PI hydrolysis by cP2Y(11) receptors (adenosine 5'-(2-O-thio) diphosphate = 2 methylthio-ADP >/= 2-methylthio-ATP >> ADP > ATP) differs from that of hP2Y(11) receptors. Microscopic examination of MDCK-D1 cells expressing carboxyl-terminal green fluorescent protein (GFP)-tagged cP2Y(11) (cP2Y(11)-GFP) receptors indicates primarily basolateral (BL) targeting. BL addition of 200 microM ADP beta S to confluent monolayers of MDCK-D1 cells produces an increase in short circuit current (I(sc)) (11.6 +/- 1.6 microA/cm(2)) whereas apical addition of agonist has no effect, confirming targeting of functional endogenous P2Y(11) receptors to the BL surface. In contrast, when either cP2Y(11) or cP2Y(11)-GFP is overexpressed in MDCK-D1 cells, the sensitivity of I(sc) to BL agonist increases by nearly 2 orders of magnitude, as if receptor density normally limited agonist potency; moreover, apical addition of ADP beta S now produces an increase in I(sc) but with low potency. The data support the BL localization of cP2Y(11) receptors and receptor coupling to changes in I(sc) in MDCK-D1 cells except in cases in which receptors are overexpressed; receptor overexpression leads to altered sensitivities and sites of coupling to physiologic responses. PMID- 11408598 TI - Prostaglandin receptor EP(4) mediates the bone anabolic effects of PGE(2). AB - Prostaglandin (PG) E(2) is a potent inducer of cortical and trabecular bone formation in humans and animals. Although the bone anabolic action of PGE(2) is well documented, the cellular and molecular mechanisms that mediate this effect remain unclear. This study was undertaken to examine the effect of pharmacological inactivation of the prostanoid receptor EP(4), one of the PGE(2) receptors, on PGE(2)-induced bone formation in vivo. We first determined the ability of EP(4)A, an EP(4)-selective ligand, to act as an antagonist. PGE(2) increases intracellular cAMP and suppresses apoptosis in the RP-1 periosteal cell line. Both effects were reversed by EP(4)A, suggesting that EP(4)A acts as an EP(4) antagonist in the cells at concentrations consistent with its in vitro binding to EP(4). We then examined the effect of EP(4) on bone formation induced by PGE(2) in young rats. Five- to 6-week-old rats were treated with PGE(2) (6 mg/kg/day) in the presence or absence of EP(4)A (10 mg/kg/day) for 12 days. We found that treatment with EP(4)A suppresses the increase in trabecular bone volume induced by PGE(2). This effect is accompanied by a suppression of bone formation indices: serum osteocalcin, extent of labeled surface, and extent of trabecular number, suggesting that the reduction in bone volume is due most likely to decreased bone formation. The pharmacological evidence presented here provides strong support for the hypothesis that the bone anabolic effect of PGE(2) in rats is mediated by the EP(4) receptor. PMID- 11408599 TI - Pituitary adenylate cyclase-activating polypeptide stimulates secretoneurin release and secretogranin II gene transcription in bovine adrenochromaffin cells through multiple signaling pathways and increased binding of pre-existing activator protein-1-like transcription factors. AB - Secretoneurin (SN) is a novel bioactive peptide that derives from the neuroendocrine protein secretogranin II (SgII) by proteolytic processing and participates in neuro-immune communication. The neuropeptide pituitary adenylate cyclase-activating polypeptide (PACAP-38) dose-dependently stimulates (EC(50) approximately 3 nM) SN release (up to 4-fold) and SgII gene expression (up to 60 fold) in cultured bovine adrenochromaffin cells. The effect of PACAP on both SN secretion and SgII mRNA levels is rapid and long lasting. We analyzed in this neuroendocrine cell model the transduction pathways involved in both SN secretion and SgII gene transcription in response to PACAP. The cytosolic calcium chelator BAPTA-AM and the nonselective calcium channel antagonist NiCl(2) equally inhibited both secretion of the peptide and transcription of the SgII gene, indicating a major contribution of calcium influx in PACAP-induced SN biosynthesis and release in chromaffin cells. Inhibition of protein kinase A (PKA) or C (PKC) also reduced PACAP-evoked SN release but did not alter the stimulatory effect of PACAP on SgII mRNA levels. Conversely, application of mitogen-activated protein kinase inhibitors suppressed PACAP-induced SgII gene expression. The effect of PACAP on SgII mRNA levels, like the effect of the PKC stimulator 12-O-tetradecanoylphorbol-13-acetate (TPA), was not affected by cycloheximide, whereas the effects of the PKA stimulator forskolin or cell depolarization by high K(+) were significantly reduced by the protein synthesis inhibitor. PACAP and TPA both increased the binding activity of the SgII cAMP response element to trans-acting factors present in chromaffin cell nuclear extracts, which are recognized by antibodies to activator protein-1-related proteins. These data indicate that SN biosynthesis is regulated by PACAP in chromaffin cells through complex signaling cascades, suggesting that SN may play a function during trans-synaptic stimulation of the adrenal medulla. PMID- 11408600 TI - Chronic exposure to mu-opioid agonists produces constitutive activation of mu opioid receptors in direct proportion to the efficacy of the agonist used for pretreatment. AB - Chronic morphine treatment has been shown to produce constitutive activation of mu-opioid receptors, and this transition might contribute to the development of tolerance and dependence. The apparent ability of chronic morphine to increase the spontaneous, agonist-independent activation of mu-opioid receptors may be unique, due to its distinct partial agonist properties of possessing a relatively high intrinsic activity coupled with a poor ability to produce desensitization and down-regulation. Therefore, the present study tested the hypothesis that prolonged exposure to morphine would produce greater constitutive activity of mu opioid receptors than exposure to the full agonist [D-Ala(2),N-MePhe(4),Gly ol(5)]enkephalin (DAMGO). GH(3) cells expressing mu-opioid receptors were exposed to chronic morphine, DAMGO, or no opioid under conditions determined to produce maximal desensitization, down-regulation, and cAMP rebound. After chronic treatment, the mu-opioid antagonists naloxone and beta-chlornaltrexamine (beta CNA) were evaluated in two assays predictive of inverse agonist activity. Both antagonists produced a concentration-dependent inhibition of [(35)S]GTP gamma S binding only in membranes prepared from cells chronically exposed to opioids. This effect was reversed by the neutral mu-opioid antagonist CTAP. Additionally, conditions known to uncouple G protein-coupled receptors from G proteins produced a leftward shift in the competition curve of beta-CNA for [(3)H]DAMGO binding only in membranes prepared from chronically treated cells. In contrast, these conditions produced no shift in the competition curve by the neutral antagonist CTAP in cells exposed to chronic DAMGO. Therefore, prolonged exposure of GH(3)MOR cells to opioids produced constitutive activation of mu-opioid receptors. Surprisingly, chronic treatment with the more efficacious agonist DAMGO produced greater increases in both measures of inverse agonist activity than did morphine. These observations may lend novel insight into the mechanisms of opioid tolerance and dependence. PMID- 11408601 TI - Nerve growth factor stimulation of p42/p44 mitogen-activated protein kinase in PC12 cells: role of G(i/o), G protein-coupled receptor kinase 2, beta-arrestin I, and endocytic processing. AB - In this study, we have shown that nerve growth factor (NGF)-dependent activation of the p42/p44 mitogen-activated protein kinase (p42/p44 MAPK) pathway in PC12 cells can be partially blocked by pertussis toxin (which inactivates the G proteins G(i/o)). This suggests that the Trk A receptor may use a G protein coupled receptor pathway to signal to p42/p44 MAPK. This was supported by data showing that the NGF-dependent activation of p42/p44 MAPK is potentiated in cells transfected with G protein-coupled receptor kinase 2 (GRK2) or beta-arrestin I. Moreover, GRK2 is constitutively bound with the Trk A receptor, whereas NGF stimulates the pertussis toxin-sensitive binding of beta-arrestin I to the TrkA receptor-GRK2 complex. Both GRK2 and beta-arrestin I are involved in clathrin mediated endocytic signaling to p42/p44 MAPK. Indeed, inhibitors of clathrin mediated endocytosis (e.g., monodansylcadaverine, concanavalin A, and hyperosmolar sucrose) reduced the NGF-dependent activation of p42/p44 MAPK. Finally, we have found that the G protein-coupled receptor-dependent component regulating p42/p44 MAPK is required for NGF-induced differentiation of PC12 cells. Thus, NGF-dependent inhibition of DNA synthesis was partially blocked by PD098059 (inhibitor of MAPK kinase-1 activation) and pertussis toxin. Our findings are the first to show that the Trk A receptor uses a classic G protein coupled receptor-signaling pathway to promote differentiation of PC12 cells. PMID- 11408602 TI - Specific activation of the alpha 7 nicotinic acetylcholine receptor by a quaternary analog of cocaine. AB - Effects of cocaine and cocaine methiodide were evaluated on the homomeric alpha 7 neuronal nicotinic receptor (nAChR). Whereas cocaine itself is a general nAChR noncompetitive antagonist, we report here the characterization of cocaine methiodide, a novel selective agonist for the alpha 7 subtype of nAChR. Data from (125)I-alpha-bungarotoxin binding assays indicate that cocaine methiodide binds to alpha 7 nAChR with a K(i) value of approximately 200 nM while electrophysiology studies indicate that the addition of a methyl group at the amine moiety of cocaine changes the drug's activity profile from inhibitor to agonist. Cocaine methiodide activates alpha 7 nAChR with an EC(50) value of approximately 50 microM and shows comparable efficacy to ACh in oocyte experiments. While agonist effects are specific for the alpha 7 neuronal nAChR and are not observed with heteromeric neuronal or skeletal muscle nAChR, antagonist effects are present for heteromeric nAChR combinations. Studies of PC12 cells transiently transfected with human alpha 7 cDNA and expressing a variety of functional nicotinic receptor subtypes confirm the specificity of cocaine methiodide agonist effects. Our results indicate that a quaternary structural derivative of cocaine can be used as a specific agonist for the alpha 7 subtype of neuronal nicotinic receptor. PMID- 11408603 TI - Complex intracellular messenger pathways regulate one type of neuronal alpha bungarotoxin-resistant nicotinic acetylcholine receptors expressed in insect neurosecretory cells (dorsal unpaired median neurons). AB - Although molecular biology provides new insights into the subunit compositions and the stoichiometries of insect neuronal nicotinic acetylcholine receptors (nAChRs), our knowledge about the phosphorylation/dephosphorylation mechanisms of native neuronal nAChRs is limited. The regulation of alpha-bungarotoxin-resistant nAChRs was studied on dissociated adult dorsal unpaired median neurons isolated from the terminal abdominal ganglion of the cockroach Periplaneta americana, using whole-cell, patch-clamp technique. Under 0.5 microM alpha-bungarotoxin treatment, pressure ejection application of nicotine or acetylcholine onto the cell body induced an inward current exhibiting a biphasic current-voltage relationship. We found that two distinct components underlying the biphasic curve differed in their ionic permeability and pharmacology (one being sensitive to d tubocurarine, and the other affected only by mecamylamine and alpha-conotoxin ImI). This indicated that two types of alpha-bungarotoxin-resistant nAChRs (named nAChR1 and nAChR2) mediated the nicotinic response. These two components were also differentially sensitive to rundown and intracellular messengers. Intracellular application of 0.1 mM cAMP only increased the current amplitude mediated by nAChR1. Using forskolin (1 microM), W7 and H89, we demonstrated that adenylyl cyclase, sensitive to calcium/calmodulin complex, regulated nAChR1 via a cAMP/cAMP-dependent protein kinase cascade. By contrast, internal cAMP concentration higher than 0.1 mM reduced the current amplitude. This effect, mimicked by high external concentration of forskolin (100 microM) and IBMX, was reversed by okadaic acid, suggesting the implication of a protein phosphatase. Using KN-62, we demonstrated that calmodulin-Kinase II also modulated directly and indirectly nAChR1, via an inhibition of the phosphatase activity. Finally, we reported that phosphorylation/dephosphorylation of nAChR1 strongly affected the action of the widely used neonicotinoid insecticide imidacloprid. PMID- 11408604 TI - Dopamine D(4) and D(2L) Receptor Stimulation of the Mitogen-Activated Protein Kinase Pathway Is Dependent on trans-Activation of the Platelet-Derived Growth Factor Receptor. AB - The ability of dopamine D(4) and D(2) receptors to activate extracellular signal regulated kinases (ERKs) 1 and 2 was compared using Chinese hamster ovary (CHO K1) cells transfected with D(4.2), D(4.4), D(4.7), and D(2L) receptors. Dopamine stimulation of D(4) or D(2L) receptors produced a transient, dose-dependent increase in ERK1/2 activity. Receptor-specific activation of the ERK mitogen activated protein kinase (MAPK) pathway was confirmed using the D(2)-like receptor-selective agonist quinpirole, whereas the specific antagonist haloperidol blocked activation. MAPK stimulation was dependent on a pertussis toxin-sensitive G protein (G(i/o)). trans-Activation of the platelet-derived growth factor (PDGF) receptor was an essential step in D(4) and D(2L) receptor induced MAPK activation. PDGF receptor-selective tyrosine kinase inhibitors tyrphostin A9 and AG1295 abolished or significantly inhibited ERK1/2 activation by D(4) and D(2L) receptors. Dopamine stimulation of the D(4) receptor also produced a rapid increase in tyrosine phosphorylation of the PDGF receptor-beta. The Src-family tyrosine kinase inhibitor PP2 blocked MAPK activation by dopamine; however, this drug was also found to inhibit PDGF-BB-stimulated ERK activity and autophosphorylation of the PDGF receptor-beta. Downstream signaling pathways support the involvement of a receptor tyrosine kinase. The phosphoinositide 3 kinase inhibitors wortmannin and LY294002, protein kinase C inhibitors GF109203X and Calphostin C, dominant-negative RasN17, and the MEK inhibitor PD98059 significantly attenuated or abolished activation of MAPK by dopamine D(4) and D(2L) receptors. Our results indicate that D(4) and D(2L) receptors activate the ERK kinase cascade by first mobilizing signaling by the PDGF receptor, followed by the subsequent activation of ERK1/2 by pathways associated with this receptor tyrosine kinase. PMID- 11408605 TI - The use of a novel taxane-based P-glycoprotein inhibitor to identify mutations that alter the interaction of the protein with paclitaxel. AB - Murine thymoma cell lines expressing mutated forms of the mdr1b P-glycoprotein were isolated using a novel taxane-based P-glycoprotein inhibitor tRA-96023 (SB RA-31012). The selection strategy required resistance to a combination of tRA 96023 and colchicine. Five mutations were identified (N350I, I862F, L865F, L868W, and A933T) that reduce the capacity of tRA-96023 to inhibit P-glycoprotein dependent drug resistance. These mutations also result in a loss of paclitaxel resistance ranging from 47 to 100%. Four mutations are located in the second half of the protein, within or near the proposed transmembrane segment (TMS) 10--11 regions. The fifth mutation (N350I) is within the first half of the protein, proximal (cytoplasmic) to TMS 6. The variant cell line expressing the L868W mutation was subjected to a second round of selection involving tRA-96023 and the toxic drug puromycin. This resulted in the isolation of a cell line expressing a P-glycoprotein with a double mutation. The additional mutation (N988D) is located within TMS 12 and conveys further decreases in resistance to paclitaxel and the capacity of tRA-96023 to inhibit drug resistance. Taken together, the results indicate a significant contribution by the TMS 10--12 portion of the protein to the recognition and transport of taxanes and give evidence that the cytoplasmic region proximal to TMS 6 also plays a role in taxane interactions with P glycoproteins. Interestingly, mutations within TMS 6 and 12 were found to cause a partial loss of PSC-833 inhibitor activity, suggesting that these regions participate in the interactions with cyclosporin and its derivatives. PMID- 11408606 TI - Reciprocal modulation of voltage-gated and background K(+) channels mediated by nucleotides and corticotropin. AB - Bovine adrenal zona fasciculata (AZF) cells express two types of K(+)-selective ion channels including a rapidly inactivating bKv1.4 current (I(A)) and an ATP dependent noninactivating background current (I(AC)) that sets the resting membrane potential. Whole-cell, patch-clamp recording from cultured AZF cells was used to demonstrate a novel reciprocal modulation of these two K(+) channels by intracellular nucleotides and corticotropin. Specifically, increases in I(AC) activity induced by intracellular ATP, as well as GTP and 5'-adenylyl imidodiphosphate (AMP-PNP), were accompanied by a corresponding decrease in the amplitude of the voltage-gated I(A) current. The reduction in I(A) current was observed only when patch pipettes contained ATP or other nucleotides at concentrations sufficient to support activation of I(AC). Conversely, the nearly complete inhibition of I(AC) by corticotropin was accompanied by the coincident reappearance of functional I(A) channels. In the absence of I(AC) current, corticotropin failed to alter I(A). The reciprocal modulation of AZF cell K(+) channels by nucleotides and corticotropin was independent of membrane voltage. These results demonstrate a new form of channel modulation in which the activity of two different K(+) channels is reciprocally modulated in tandem through hormonal and metabolic signaling pathways. They further suggest that I(A) and I(AC) K(+) channels may be functionally coupled in a dynamic equilibrium driven by intracellular ATP and G-protein-coupled receptors. This may represent a unique mechanism for transducing biochemical signals to ionic events involved in cortisol secretion. PMID- 11408607 TI - The peptide YY-preferring receptor mediating inhibition of small intestinal secretion is a peripheral Y(2) receptor: pharmacological evidence and molecular cloning. AB - A peptide YY (PYY)-preferring receptor [PYY > neuropeptide Y (NPY)] was previously characterized in rat small intestinal crypt cells, where it mediates inhibition of fluid secretion. Here, we investigated the possible status of this receptor as a peripheral Y(2) receptor in rats. Typical Y(2) agonists (PYY(3-36), NPY(3-36), NPY(13-36), C2-NPY) and very short PYY analogs (N-alpha-Ac-PYY(22-36) and N-alpha-Ac-PYY(25-36)) acting at the intestinal PYY receptor were tested for their ability to inhibit the binding of (125)I-PYY to membranes of rat intestinal crypt cells and of CHO cells stably transfected with the rat hippocampal Y(2) receptor cDNA. Similar PYY preference was observed and all analogs exhibited comparable high affinity in both binding assays. The same held true for the specific Y(2) antagonist BIIE0246 with a K(i) value of 6.5 and 9.0 nM, respectively. BIIE0246 completely abolished the inhibition of cAMP production by PYY in crypt cells and transfected CHO cells. Moreover, the antagonist 1) considerably reversed the PYY-induced reduction of short-circuit current in rat jejunum mucosa in Ussing chamber and 2) completely abolished the antisecretory action of PYY on vasoactive intestinal peptide (VIP)-induced fluid secretion in rat jejunum in vivo. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) experiments showed that Y(2) receptor transcripts were present in intestinal crypt cells (3 x 10(2) molecules/100 ng RNA(T)) with no expression in villus cells, in complete agreement with the exclusive binding of PYY in crypt cells. Finally, a full-length Y(2) receptor was cloned by RT-PCR from rat intestinal crypt cells and also from human small intestine. We conclude that the so-called PYY-preferring receptor mediating inhibition of intestinal secretion is a peripheral Y(2) receptor. PMID- 11408608 TI - Immune-specific up-regulation of adseverin gene expression by 2,3,7,8 tetrachlorodibenzo-p-dioxin. AB - To identify genes that are regulated by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and possibly involved in TCDD-induced immunotoxicity, we used the differential display technique to screen for differentially expressed genes in the mouse thymus. Here we show that TCDD increased the expression of adseverin, a Ca(2+)-dependent, actin-severing protein. The induction of adseverin is dose- and time-dependent in parallel with the induction of CYP1A1, which is currently the most frequently used marker for TCDD exposure. A comparison between mouse strains with different TCDD responsiveness indicated that the induction of adseverin is dependent on the aryl hydrocarbon receptor, a transcription factor known to mediate most of TCDD's biological effects. Examination of additional organs revealed that the up-regulation of the adseverin gene expression is immune specific. Using an anti-adseverin antibody, we confirmed the induction of adseverin by TCDD at the protein level and it was confined to the thymic cortex, which harbors immature thymocytes that are known target cells of TCDD. Considering adseverin's role in actin cytoskeletal reorganization, our observations reveal new mechanistic aspects of how TCDD might exert some of its immunotoxic effects. PMID- 11408609 TI - Sequence-dependent potentiation of paclitaxel-mediated apoptosis in human leukemia cells by inhibitors of the mitogen-activated protein kinase kinase/mitogen-activated protein kinase pathway. AB - Effects of inhibitors of the mitogen-activated protein kinase kinase/mitogen activated protein kinase (MEK/MAPK) cascade have been examined in relation to paclitaxel-induced apoptosis in human monocytic leukemia cells (U937). Cells treated with paclitaxel (250 nm; 6 h) followed by PD98059 [corrected] exhibited a significant increase in mitochondrial dysfunction (e.g., cytochrome c release), caspase activation, poly ADP-ribose polymerase cleavage, and apoptosis, whereas pretreatment of cells with PD98059 reduced lethality. Similar results were obtained with other MEK/MAPK inhibitors (e.g., U0126 and PD184352). Subsequent exposure of paclitaxel-treated cells to PD98059 did not enhance dephosphorylation/activation of p34(cdc2) but diminished expression of the antiapoptotic protein Mcl-1. The caspase inhibitor ZVAD-fmk opposed potentiation of paclitaxel-induced loss of mitochondrial membrane potential (Deltapsi(m)) and apoptosis by PD98059, but not cytochrome c release. Paclitaxel treatment induced sustained phosphorylation/activation of MAPK, an effect prevented by subsequent, but not prior, exposure to PD98059. Paclitaxel treatment also induced c-Jun N terminal kinase phosphorylation, but this effect was enhanced only slightly by subsequent PD98059 administration. Although paclitaxel alone failed to induce p38 MAPK activation, subsequent (but not prior) exposure to PD98059 induced a dramatic increase in p38 MAPK phosphorylation. Moreover, coadministration of the p38 MAPK inhibitors SB203580 and SB202190 abrogated the increase in paclitaxel mediated apoptosis induced by PD98059. Finally, subsequent PD98059 exposure increased, whereas prior exposure decreased inhibition of clonogenicity by paclitaxel. Together, these findings suggest that subsequent exposure of paclitaxel-treated U937 cells to MEK/MAPK inhibitors induces perturbations in signaling pathways, particularly the p42/44 MAPK and p38 MAPK cascades, that lower the threshold for mitochondrial injury and induction of cell death. PMID- 11408610 TI - Evidence for a new G protein-coupled cannabinoid receptor in mouse brain. AB - The purpose of these studies was to support the hypothesis that an undiscovered cannabinoid receptor exists in brain. [(35)S]GTP gamma S binding was stimulated by anandamide and WIN55212-2 in brain membranes from both CB(1)(+/+) and CB(1)(-/ ) mice. In contrast, a wide variety of other compounds that are known to activate CB(1) receptors, including CP55940, HU-210, and Delta(9)-tetrahydrocannabinol, failed to stimulate [(35)S]GTP gamma S binding in CB(1)(-/-) membranes. In CB(1)( /-) membranes, SR141716A affected both basal and anandamide- or WIN55212-2 induced stimulation of [(35)S]GTP gamma S binding only at concentrations greater than 1 microM. In CB(1)(+/+) membranes, SR141716A inhibited only 84% of anandamide and 67% of WIN55212-2 stimulated [(35)S]GTP gamma S binding with an affinity appropriate for mediation by CB(1) receptors (K(B) approximately 0.5 nM). The remaining stimulation seemed to be inhibited with lower potency (IC(50) approximately 5 microM) similar to that seen in CB(1)(-/-) membranes or in the absence of agonist. Further experiments determined that the effects of anandamide and WIN55212-2 were not additive, but that the effect of mu opioid, adenosine A1, and cannabinoid ligands were additive. Finally, assays of different central nervous system (CNS) regions demonstrated significant activity of cannabinoids in CB(1)(-/-) membranes from brain stem, cortex, hippocampus, diencephalon, midbrain, and spinal cord, but not basal ganglia or cerebellum. Moreover, some of these same CNS regions also showed significant binding of [(3)H]WIN55212-2, but not [(3)H]CP55940. Thus anandamide and WIN55212-2 seemed to be active in CB(1)(-/ ) mouse brain membranes via a common G protein-coupled receptor with a distinct CNS distribution, implying the existence of an unknown cannabinoid receptor subtype in brain. PMID- 11408611 TI - Mutation of Asp(171) and Asp(262) of the chemokine receptor CXCR4 impairs its coreceptor function for human immunodeficiency virus-1 entry and abrogates the antagonistic activity of AMD3100. AB - The bicyclam AMD3100 is a highly potent and selective CXCR4 antagonist with strong antiviral activity against human immunodeficiency virus (HIV)-1 and HIV-2, which use CXCR4 as coreceptor for host cell entry. Here, we investigated the interaction of AMD3100 with CXCR4 at the molecular level by mutational analysis. We established a set of stably transfected U87.CD4 cell lines expressing different mutant forms of CXCR4 (i.e., CXCR4[WT], CXCR4[D171N], CXCR4[D262N], CXCR4[D171N,D262N], and CXCR4[H281A]), to compare the activity of the compound against mutated versus wild-type CXCR4. We found that the antagonistic action of AMD3100 against CXCR4--as assessed by the inhibitory effects of the compound on stromal cell-derived factor (SDF-1) binding to its receptor and on SDF-1-induced intracellular calcium signaling, and by displacement of the CXCR4-specific antibody, clone 12G5--was greatly reduced by substitution of Asp(171) and/or Asp(262) by neutral asparagine residue(s). Both aspartates, but most particularly Asp(262), also proved essential for the anti-HIV-1 activity of AMD3100 against the viruses NL4.3, IIIB, and HE. In contrast, substitution of His(281) by a neutral alanine potentiated the antagonistic and antiviral effects of the compound in the different assay systems. Importantly, compared with the wild-type receptor, CXCR4[D262N] was much less effective, whereas CXCR4[D171N,D262N] completely failed as a coreceptor for infection by HIV-1 NL4.3. Thus, the negatively charged aspartate residues at positions 171 and 262, located in transmembrane domains 4 and 6 of the 7-transmembrane receptor, respectively, may represent crucial sites for electrostatic interaction of the positive charges of the bicyclams, as well as for the highly basic V3 loop of the gp120 envelope protein of certain HIV-1 strains. PMID- 11408612 TI - c-Myc down-regulation increases susceptibility to cisplatin through reactive oxygen species-mediated apoptosis in M14 human melanoma cells. AB - Our aim in this work was to define the role of c-Myc in the susceptibility to cisplatin [cis-diamminedichloroplatinum(II) (CDDP)] in human melanoma cells. Two M14 melanoma cell clones obtained by transfection and expressing six to ten times lower c-Myc protein levels than the parental cells and the control clone were employed. Analysis of survival curves demonstrates an increase in CDDP sensitivity in c-Myc low-expressing clones if compared with the control clone and the parental line. The enhanced sensitivity is unrelated to the impairment in enzymatic DNA repair activity. Cell cycle analysis demonstrates that although the control clone is able to completely recover from the CDDP-induced S-G(2)/M block, this arrest is prolonged in c-Myc low-expressing clones and a fraction of cells undergoes apoptosis. Although no changes in P53, Bax, Bcl-2, and Bcl-x(L/S) protein levels are observed, apoptosis is associated with the formation of reactive oxygen species (ROS), activation of caspase-1, caspase-3 and cleavage of the specific caspase substrate poly-ADP-ribose polymerase. The use of the antioxidant N-acetyl cysteine and caspase inhibitors prevents CDDP-induced apoptosis in c-Myc low-expressing clones, demonstrating that ROS, caspase-1, and caspase-3 are required for apoptotic cell death. Moreover, ROS generation depends on caspase-1-like activation because the Ac-YVAD-cho inhibitor abrogates CDDP induced ROS in the c-Myc low-expressing clones. PMID- 11408613 TI - Memantine inhibits efferent cholinergic transmission in the cochlea by blocking nicotinic acetylcholine receptors of outer hair cells. AB - Memantine is a blocker of Ca(2+)-permeable glutamate and nicotinic acetylcholine receptors (nAChR). We investigated the action of memantine on cholinergic synaptic transmission at cochlear outer hair cells (OHCs). At this inhibitory synapse, hyperpolarization of the postsynaptic cell results from opening of SK type Ca(2+)-activated K(+) channels via a highly Ca(2+)-permeable nAChR containing the alpha 9 subunit. We show that inhibitory postsynaptic currents recorded from OHCs were reversibly blocked by memantine with an IC(50) value of 16 microM. RT-PCR revealed that a newly cloned nAChR subunit, alpha 10, is expressed in OHCs. In contrast to homomeric expression, coexpression of alpha 9 and alpha 10 subunits in Xenopus laevis oocytes resulted in robust acetylcholine induced currents, indicating that the OHC nAChR may be an alpha 9/alpha 10 heteromer. Accordingly, nAChR currents evoked by application of the ligand to OHCs and currents through alpha 9/alpha 10 were blocked by memantine with a similar IC(50) value of about 1 microM. Memantine block of alpha 9/alpha 10 was moderately voltage dependent. The lower efficacy of memantine for inhibition of inhibitory postsynaptic currents (IPSCs) most probably results from a blocking rate that is slow with respect to the short open time of the receptor channels during an IPSC. Thus, synaptic transmission in OHCs is inhibited by memantine block of Ca(2+) influx through nAChRs. Importantly, prolonged receptor activation and consequently massive Ca(2+) influx, as might occur under pathological conditions, is blocked at low micromolar concentrations, whereas the fast IPSCs initiated by short receptor activation are only blocked at concentrations above 10 microM. PMID- 11408614 TI - Characterization of a mutant sulfonylurea receptor SUR2B with high affinity for sulfonylureas and openers: differences in the coupling to Kir6.x subtypes. AB - ATP-dependent K(+) channels are composed of pore-forming subunits of the Kir6.x family and of sulfonylurea receptors (SURs). SUR1, expressed in pancreatic beta cells, has a higher affinity for sulfonylureas, such as glibenclamide, than SUR2B, expressed in smooth muscle. This difference is mainly caused by serine 1237 in SUR1 corresponding to tyrosine 1206 in SUR2B. To increase the affinity of SUR2B for glibenclamide, the mutant SUR2B(Y1206S) was constructed. In whole-cell patch-clamp experiments, glibenclamide inhibited the channel formed by coexpression of mutant SUR2B with Kir6.1 or 6.2 in human embryonic kidney cells with IC(50) values of 2.7 and 13 nM, respectively (wild-type, 43 and 167 nM). In intact cells, [(3)H]glibenclamide bound to mutant SUR2B with a K(D) value of 4.7 nM (wild-type, 32 nM); coexpression with Kir6.1 or 6.2 increased affinity by 4- and 8-fold, respectively. Binding of the opener [(3)H]P1075 to SUR2B(Y1206S) was the same as to wild-type and was unaffected by coexpression. In cells, the ratio of glibenclamide:P1075 sites was approximately 1:1; in membranes, it varied with the MgATP concentration. Heterologous competition curves were generally biphasic; the shape of the curve depended on the Kir-subtype. The effects of coexpression were weakened or abolished when binding assays were conducted in membranes. It is concluded that the mutation Y1206S increases the affinity of SUR2B for and the channel sensitivity toward glibenclamide by 7- to 15-fold. The interaction of glibenclamide (but not opener) with mutant SUR2B is modified by coexpression with Kir6.x in a manner depending on the Kir subtype and on the integrity of the cell. PMID- 11408615 TI - Fast and slow gating of CLC-1: differential effects of 2-(4-chlorophenoxy) propionic acid and dominant negative mutations. AB - Our knowledge about ClC-1 muscle chloride channel gating, previously gained from single-channel recording and noise analysis, provides a theoretical basis for further analysis of macroscopic currents. In the present study, we propose a simple method of calculation of open probabilities (P(o)) of fast and slow gates from the relative amplitudes of ClC-1 inward current components. With this method, we investigated the effects of 2-(4-chlorophenoxy) propionic acid (CPP), a drug known to produce myotonia in animals, and dominant negative myotonic mutations, F307S and A313T, on fast and slow gating of ClC-1. We have shown that these mutations affected the P(o) of the slow gate, as expected from their mode of inheritance, and that CPP predominantly affected the fast gating process. CPP's action on the fast gating of mutant channels was similar to its effect in wild-type channels. Comparison of the effects of CPP and the mutations on fast and slow gating with the effects produced by reduction of external Cl(-) concentration suggested that CPP and mutations exert their action by affecting the transition of the channel from its closed to open state after Cl(-) binding to the gating site. PMID- 11408616 TI - Role of nitric oxide in down-regulation of CYP2B1 protein, but not RNA, in primary cultures of rat hepatocytes. AB - There are conflicting reports about the role of nitric oxide in the down regulation of cytochrome P450 that occurs when animals or cultured hepatocytes are exposed to inflammatory stimuli. Here, we investigated the participation of NO in the down-regulation of CYP2B1 by bacterial endotoxin (LPS) in rat hepatocytes cultured on Matrigel. LPS caused the down-regulation of CYP2B1 mRNA to 20% of control values within 12 h of treatment, and this was not reversed by concentrations of NO synthase inhibitors that completely blocked NO production. LPS was several orders of magnitude more potent in the down-regulation of CYP2B1 mRNA than in induction of NO production. In contrast, concentrations of LPS in the 1 to 100 ng/ml range induced NO production and produced a rapid down regulation of CYP2B1 protein to 30% and <5% of control at 6 and 24 h, respectively, that could be completely prevented both by inhibitors of NO synthase and by LY83583, which prevents NO synthase-2 induction. The blockade of CYP2B1 down-regulation by NO synthase inhibitors was reversed by arginine, and the NO donors S-nitrosoglutathione and S-nitroso-N-acetylpenicillamine mimicked CYP2B1 protein suppression. Taken together, these experiments demonstrate two independent mechanisms of CYP2B1 down-regulation by LPS: a rapid, NO-dependent suppression of the protein occurring at high concentrations of LPS and a slower, NO-independent pretranslational suppression occurring at low concentrations of LPS. PMID- 11408617 TI - Resveratrol inhibits phorbol ester and UV-induced activator protein 1 activation by interfering with mitogen-activated protein kinase pathways. AB - Resveratrol, a phenolic compound found in grapes and other food products, prevents chemical-induced carcinogenesis in a number of animal models of cancers. To better understand its chemopreventive property, we examined effects of resveratrol on the activity of activator protein 1 (AP-1), a dimeric transcription factor that plays a critical role in the carcinogenesis and tumor transformation. Pretreatment of HeLa cells with resveratrol inhibited the transcription of AP-1 reporter gene by UVC and phorbol 12-myristate 13-acetate (PMA). Pretreatment with resveratrol also inhibited the activation of extracellular signal-regulated protein kinase 2 (ERK2), c-jun N-terminal kinase 1 (JNK1), and p38. Selectively blocking mitogen-activated protein kinase (MAPK) pathways by overexpression of dominant-negative mutants of kinases attenuated the AP-1 activation by PMA and UVC. Interestingly, resveratrol had little effect on the induction of AP-1 reporter gene by active Raf-1, MEKK1, or MKK6, suggesting that it inhibited MAPK pathways by targeting the signaling molecules upstream of Raf-1 or MEKK1. Indeed, incubation of resveratrol with the isolated c-Src protein tyrosine kinase and protein kinase C diminished their kinase activities. Furthermore, inhibition of protein tyrosine kinases and protein kinase C with their selective inhibitors impaired the activation of MAPKs as well as the induction of AP-1 activity by PMA and UVC. In addition, modulation of estrogen receptor activity with 17beta-estradiol had no effect on the inhibition of AP-1 by resveratrol. Taken together, these results suggest that the effects of resveratrol on AP-1 and MAPK pathways may involve the inhibition of both protein tyrosine kinases and protein kinase C. PMID- 11408618 TI - Mechanism of mitotic block and inhibition of cell proliferation by the semisynthetic Vinca alkaloids vinorelbine and its newer derivative vinflunine. AB - The two second-generation Vinca alkaloids, vinorelbine and vinflunine, affect microtubule dynamics very differently from vinblastine, a first generation Vinca alkaloid. For example, vinblastine strongly suppresses the rate and extent of microtubule shortening in vitro, whereas vinorelbine and vinflunine suppress the rate and extent of microtubule growing events. We asked whether these differences result in differences in mitotic spindle organization that might be responsible for the superior antitumor activities of the two second-generation Vinca alkaloids. IC(50) values for inhibition of HeLa cell proliferation for vinflunine, vinorelbine, and vinblastine were 18, 1.25, and 0.45 nM, respectively, similar to the concentrations that induced mitotic block at the metaphase/anaphase transition (38, 3.8, and 1.1 nM, respectively), indicating that mitotic block is a major contributor to antiproliferative action for all three drugs. Mitotically blocked cells exhibited aberrant spindles, consistent with induction of block by suppression of microtubule dynamics. Despite differences in their actions on individual dynamic instability parameters, morphologically detectable differences in spindle effects among the three drugs were minimal, indicating that overall suppression of dynamics may be more important in blocking mitosis than specific effects on growth or shortening. We also found that the peak intracellular drug concentration at the mitotic IC(50) value was highest for vinflunine (4.2 +/- 0.2 microM), intermediate for vinorelbine (1.3 +/- 0.1 microM), and more than 10-fold lower for vinblastine (130 +/- 7 nM), suggesting that intracellular binding reservoir(s) may be partially responsible for vinflunine's high efficacy and minimal side effects. PMID- 11408619 TI - Loss of huntingtin-mediated BDNF gene transcription in Huntington's disease. AB - Huntingtin is a 350-kilodalton protein of unknown function that is mutated in Huntington's disease (HD), a neurodegenerative disorder. The mutant protein is presumed to acquire a toxic gain of function that is detrimental to striatal neurons in the brain. However, loss of a beneficial activity of wild-type huntingtin may also cause the death of striatal neurons. Here we demonstrate that wild-type huntingtin up-regulates transcription of brain-derived neurotrophic factor (BDNF), a pro-survival factor produced by cortical neurons that is necessary for survival of striatal neurons in the brain. We show that this beneficial activity of huntingtin is lost when the protein becomes mutated, resulting in decreased production of cortical BDNF. This leads to insufficient neurotrophic support for striatal neurons, which then die. Restoring wild-type huntingtin activity and increasing BDNF production may be therapeutic approaches for treating HD. PMID- 11408620 TI - The human nuclear xenobiotic receptor PXR: structural determinants of directed promiscuity. AB - The human nuclear pregnane X receptor (hPXR) activates cytochrome P450-3A expression in response to a wide variety of xenobiotics and plays a critical role in mediating dangerous drug-drug interactions. We present the crystal structures of the ligand-binding domain of hPXR both alone and in complex with the cholesterol-lowering drug SR12813 at resolutions of 2.5 and 2.75 angstroms, respectively. The hydrophobic ligand-binding cavity of hPXR contains a small number of polar residues, permitting SR12813 to bind in three distinct orientations. The position and nature of these polar residues were found to be critical for establishing the precise pharmacologic activation profile of PXR. Our findings provide important insights into how hPXR detects xenobiotics and may prove useful in predicting and avoiding drug-drug interactions. PMID- 11408621 TI - Tauopathy in Drosophila: neurodegeneration without neurofibrillary tangles. AB - The microtubule-binding protein tau has been implicated in the pathogenesis of Alzheimer's disease and related disorders. However, the mechanisms underlying tau mediated neurotoxicity remain unclear. We created a genetic model of tau-related neurodegenerative disease by expressing wild-type and mutant forms of human tau in the fruit fly Drosophila melanogaster. Transgenic flies showed key features of the human disorders: adult onset, progressive neurodegeneration, early death, enhanced toxicity of mutant tau, accumulation of abnormal tau, and relative anatomic selectivity. However, neurodegeneration occurred without the neurofibrillary tangle formation that is seen in human disease and some rodent tauopathy models. This fly model may allow a genetic analysis of the cellular mechanisms underlying tau neurotoxicity. PMID- 11408622 TI - Deterministic delivery of a single atom. AB - We report the realization of a deterministic source of single atoms. A standing wave dipole trap is loaded with one or any desired number of cold cesium atoms from a magneto-optical trap. By controlling the motion of the standing wave, we adiabatically transport the atom with submicrometer precision over macroscopic distances on the order of a centimeter. The displaced atom is observed directly in the dipole trap by fluorescence detection. The trapping field can also be accelerated to eject a single atom into free flight with well-defined velocities. PMID- 11408623 TI - Questions regarding Precambrian sulfur isotope fractionation. PMID- 11408624 TI - Earth system science. PMID- 11408625 TI - Global warming. Bush backs spending for a 'global problem'. PMID- 11408626 TI - Underground laboratory. U.S. researchers go for scientific gold mine. PMID- 11408627 TI - Nazi research. Max Planck offers historic apology. PMID- 11408628 TI - DNA sequencing. Genome teams adjust to shotgun marriage. PMID- 11408629 TI - Neurodegenerative disease. Using the fruit fly to model tau malfunction. PMID- 11408630 TI - Espionage case. Japan says cell lines weren't used at RIKEN. PMID- 11408631 TI - Astronomy. Infrared gleam stamps brown dwarfs as stars. PMID- 11408632 TI - Astrophysics. Quasars or blazars? It's all in the angle. PMID- 11408633 TI - Animal models. EC boosts funds for mutant mice. PMID- 11408634 TI - Condensed-matter physics. Switch-hitter materials tantalize theorists. PMID- 11408635 TI - The future of NIH. Who will be custodian of the crown jewels? PMID- 11408636 TI - The people. Ruth Kirschstein and Alan Rabson. PMID- 11408637 TI - The big three. NCI's Richard Klausner. PMID- 11408638 TI - The big three. NIAID's Anthony Fauci. PMID- 11408639 TI - The big three. NHLBI's Claude Lenfant. PMID- 11408640 TI - The numbers. NIH prays for a soft landing after its doubling ride ends. PMID- 11408641 TI - The numbers. NIH stays the course in choosing how to spend its growing budget. PMID- 11408642 TI - The winners. Even in a time of plenty, some do better than others. PMID- 11408643 TI - Medicine. Placebo-controls in short-term clinical trials of hypertension. PMID- 11408644 TI - Transcription. Translocating tubby. PMID- 11408645 TI - Chemistry. A new twist on chirality. PMID- 11408646 TI - Plant biology. One for all? PMID- 11408647 TI - Biochemistry. How to make a superior cell. PMID- 11408648 TI - Atmospheric science. Reshaping the theory of cloud formation. PMID- 11408649 TI - Planetary science. Life without photosynthesis. PMID- 11408650 TI - Discovery of hidden blazars. AB - Every radio-loud quasar may have blazar activities, according to a unified scheme where the differences in both optical and radio observations of radio-loud quasars are the result of different viewing angles. We have predicted that blazars may be detected using emission line ratio variations caused by variable illumination of gas clouds in the broad emission line region. In a spectroscopic search of 62 quasars at a redshift of about 2, we have discovered large (>20%) variations of the emission line ratios, CIV/CIII] or CIV/Lyalpha, when compared with data taken more than 10 years ago. This result is consistent with our prediction and supports the unification scheme for radio-loud quasars. PMID- 11408651 TI - Observation of magnetic hysteresis at the nanometer scale by spin-polarized scanning tunneling spectroscopy. AB - Using spin-polarized scanning tunneling microscopy in an external magnetic field, we have observed magnetic hysteresis on a nanometer scale in an ultrathin ferromagnetic film. An array of iron nanowires, being two atomic layers thick, was grown on a stepped tungsten (110) substrate. The microscopic sources of hysteresis in this system-domain wall motion, domain creation, and annihilation were observed with nanometer spatial resolution. A residual domain 6.5 nanometers by 5 nanometers in size has been found which is inherently stable in saturation fields. Its stability is the consequence of a 360 degrees spin rotation. With magnetic memory bit sizes approaching the superparamagnetic limit with sub-10 nanometer characteristic lengths, the understanding of the basic physical phenomena at this scale is of fundamental importance. PMID- 11408652 TI - Far-reaching effects of the Hawaiian Islands on the Pacific Ocean-atmosphere system. AB - Using satellite data, we detected a wind wake trailing westward behind the Hawaiian Islands for 3000 kilometers, a length many times greater than observed anywhere else on Earth. This wind wake drives an eastward ocean current that draws warm water from the Asian coast 8000 kilometers away, leaving marked changes in surface and subsurface ocean temperature. Standing in the path of the steady trade winds, Hawaii triggers an air-sea interaction that provides the feedback to sustain the influence of these small islands over a long stretch of the Pacific Ocean. PMID- 11408653 TI - Chiral sign induction by vortices during the formation of mesophases in stirred solutions. AB - Achiral diprotonated porphyrins, forming homoassociates in aqueous solution, lead to spontaneous chiral symmetry breaking. The unexpected result is that the chirality sign of these homoassociates can be selected by vortex motion during the aggregation process. This result is confirmed by means of circular dichroism spectra. These experimental findings are rationalized in terms of the asymmetric influence of macroscopic forces on bifurcation scenarios and by considering the specific binding characteristics of the porphyrin units to form the homoassociates. PMID- 11408654 TI - Modulation of cell proliferation by heterotrimeric G protein in Arabidopsis. AB - The alpha subunit of a prototypical heterotrimeric GTP-binding protein (G protein), which is encoded by a single gene (GPA1) in Arabidopsis, is a modulator of plant cell proliferation. gpa1 null mutants have reduced cell division in aerial tissues throughout development. Inducible overexpression of GPA1 in Arabidopsis confers inducible ectopic cell division. GPA1 overexpression in synchronized BY-2 cells causes premature advance of the nuclear cycle and the premature appearance of a division wall. Results from loss of function and ectopic expression and activation of GPA1 indicate that this subunit is a positive modulator of cell division in plants. PMID- 11408655 TI - G protein regulation of ion channels and abscisic acid signaling in Arabidopsis guard cells. AB - The phytohormone abscisic acid (ABA) promotes plant water conservation by decreasing the apertures of stomatal pores in the epidermis through which water loss occurs. We found that Arabidopsis thaliana plants harboring transferred DNA insertional mutations in the sole prototypical heterotrimeric GTP-binding (G) protein alpha subunit gene, GPA1, lack both ABA inhibition of guard cell inward K(+) channels and pH-independent ABA activation of anion channels. Stomatal opening in gpa1 plants is insensitive to inhibition by ABA, and the rate of water loss from gpa1 mutants is greater than that from wild-type plants. Manipulation of G protein status in guard cells may provide a mechanism for controlling plant water balance. PMID- 11408656 TI - Trophic conversion of an obligate photoautotrophic organism through metabolic engineering. AB - Most microalgae are obligate photoautotrophs and their growth is strictly dependent on the generation of photosynthetically derived energy. We show that the microalga Phaeodactylum tricornutum can be genetically engineered to thrive on exogenous glucose in the absence of light through the introduction of a gene encoding a glucose transporter (glut1 or hup1). This demonstrates that a fundamental change in the metabolism of an organism can be accomplished through the introduction of a single gene. This also represents progress toward the use of fermentation technology for large-scale commercial exploitation of algae by reducing limitations associated with light-dependent growth. PMID- 11408657 TI - Telomere position effect in human cells. AB - In yeast, telomere position effect (TPE) results in the reversible silencing of genes near telomeres. Here we demonstrate the presence of TPE in human cells. HeLa clones containing a luciferase reporter adjacent to a newly formed telomere express 10 times less luciferase than do control clones generated by random integration. Luciferase expression is restored by trichostatin A, a histone deacetylase inhibitor. Overexpression of a human telomerase reverse transcriptase complementary DNA results in telomere elongation and an additional 2- to 10-fold decrease in expression in telomeric clones but not control clones. The dependence of TPE on telomere length provides a mechanism for the modification of gene expression throughout the replicative life-span of human cells. PMID- 11408658 TI - Ordering genes in a flagella pathway by analysis of expression kinetics from living bacteria. AB - The recent advances in large-scale monitoring of gene expression raise the challenge of mapping systems on the basis of kinetic expression data in living cells. To address this, we measured promoter activity in the flagellar system of Escherichia coli at high accuracy and temporal resolution by means of reporter plasmids. The genes in the pathway were ordered by analysis algorithms without dependence on mutant strains. The observed temporal program of transcription was much more detailed than was previously thought and was associated with multiple steps of flagella assembly. PMID- 11408659 TI - Vitamin C-induced decomposition of lipid hydroperoxides to endogenous genotoxins. AB - Epidemiological data suggest that dietary antioxidants play a protective role against cancer. This has led to the proposal that dietary supplementation with antioxidants such as vitamin C (vit C) may be useful in disease prevention. However, vit C has proved to be ineffective in cancer chemoprevention studies. In addition, concerns have been raised over potentially deleterious transition metal ion-mediated pro-oxidant effects. We have now determined that vit C induces lipid hydroperoxide decomposition to the DNA-reactive bifunctional electrophiles 4-oxo 2-nonenal, 4,5-epoxy-2(E)-decenal, and 4-hydroxy-2-nonenal. The compound 4,5 Epoxy-2(E)-decenal is a precursor of etheno-2'-deoxyadenosine, a highly mutagenic lesion found in human DNA. Vitamin C-mediated formation of genotoxins from lipid hydroperoxides in the absence of transition metal ions could help explain its lack of efficacy as a cancer chemoprevention agent. PMID- 11408660 TI - From genome to function. PMID- 11408661 TI - Rb5Mo27Se31, a novel ternary reduced molybdenum selenide containing Mo12 and Mo15 clusters. AB - The crystal structure of Rb5Mo27Se31, pentarubidium heptacosamolybdenum hentriacontaselenium, consists of a mixture of Mo12Se14Se6 and Mo15Se17Se6 cluster units in a 1:1 ratio. Both types of cluster are interconnected through inter-unit Mo-Se bonds. Rb+ cations occupy large voids between the different cluster units. PMID- 11408662 TI - Hexagonal high-temperature form of aluminium phosphate tridymite from X-ray powder data. AB - Similar to silica tridymite, AlPO4 tridymite shows a sequence of displacive phase transitions resulting in a dynamically disordered hexagonal high-temperature modification. Rietveld refinement reveals that the thermal motions of the tetrahedra can be described either by strongly anisotropic displacement parameters for oxygen or by split O atoms. Due to the ordered distribution of aluminium and phosphorus over alternating tetrahedra, the space group symmetry of high-temperature AlPO4 tridymite is reduced with respect to SiO2 tridymite from P6(3)/mmc to P6(3)mc. PMID- 11408663 TI - Lu2SiO5 by single-crystal X-ray and neutron diffraction. AB - The structure of dilutetium silicon pentaoxide, Lu2SiO5, has isolated ionic SiO4 tetrahedral units and non-Si-bonded O atoms in distorted OLu4 tetrahedra. The OLu4 tetrahedra form edge-sharing infinite chains and double O2Lu6 tetrahedra along the c axis. The edge-sharing chains are connected to the O2Lu6 double tetrahedra by isolated SiO4 units. The structure has been determined by neutron diffraction. PMID- 11408664 TI - Chromous hydrazine sulfate. AB - The title compound, catena-poly[dihydraziniochromium(II)-di-mu-sulfato-O:O'], [Cr(N2H5)2(SO4)2], was studied at 100 K and contains chains of chromium(II) ions linked by pairs of sulfate anions and coordinated to hydrazinium ions. The unique Cr atom lies on an inversion centre. All five H atoms were experimentally located and are involved in hydrogen bonding to O atoms of the sulfate groups. PMID- 11408665 TI - catena-Poly[dibromozinc(II)-mu-ethylenediamine-N:N']: resolution of structural anomalies resulting from the interpretation of vibrational spectra. AB - The title structure, [ZnBr2(C2H8N2)], is made up of infinite -ZnBr2-(en)-ZnBr2 (en)- zigzag chains. Each repeat unit contains a trans ethylenediamine ligand [N C-C-N -179 (1) degrees ], which bridges two approximately tetrahedral but crystallographically distinct Zn atoms. One Zn atom is bisected by a crystallographic twofold axis, whereas the other has mirror symmetry. Even though the crystal packing does not allow significant interaction between Zn and N atoms on adjacent chains, it does facilitate extensive intermolecular N-H...Br hydrogen bonding (N...H 2.69-2.96 A). PMID- 11408666 TI - Dichlorobis(dibenzylamino)bis(tetrahydrofuran)zirconium(IV) toluene hemisolvate. AB - The title complex, [ZrCl2(C4H8O)2(C14H14N)2].0.5C7H8, was prepared in an unusual manner by utilizing [Mg[N(CH2Ph)2]2] as a ligand transfer reagent. The Zr atom lies in a distorted octahedral environment where steric repulsion from the large dibenzylamino ligands leads to a widening of the N-Zr-N angle [99.95 (9) degrees ] and corresponding compression of other angles [Cl-Zr-Cl 160.95 (3) degrees and O-Zr-O 78.22 (7) degrees ]. This distortion is compared with those found in the previously determined structures of the dimethylamino and diethylamino analogues. PMID- 11408667 TI - catena-Poly[europium-tri-mu-4-methyl-benzoato]. AB - Each Eu3+ ion in the title compound, catena-poly[europium(III)-tri-mu-4 methylbenzoato-O:O,O';O:O,O';O,O':O'], [[Eu(C8H7O2)3]3]n, is coordinated by nine O atoms, and three Eu atoms form a trimeric unit. These trimeric units are linked by bridging-chelating carboxylates to form an infinite one-dimensional polymer chain. PMID- 11408668 TI - cis-Tetracarbonyl[4-fluorophenyl)-diphenylphosphine-P](piperidine-N) molybdenum(0). AB - The title molecule, [Mo[P(C6H5)2(C6H4F)](HNC5H10)(CO)4] or [Mo(C18H14FP)(C5H11N)(CO)4], has irregular octahedral geometry about the Mo atom. The molecules form a complicated hydrogen-bonded network comprising C-H...O, C H...F and C-H...pi hydrogen bonds and pi-pi interactions. The C-H...pi and pi-pi interactions form chains containing C-H...pi/pi-pi dimers linked via C-H...pi interactions and the chains are linked into a three-dimensional network via C H...O and C-H...F hydrogen bonds. PMID- 11408669 TI - Bis[N-(6-amino-3,4-dihydro-3-methyl-5-nitroso-4-oxypyrimidin-2-yl)glycyl glycinato]triaquacalcium: coordination polymer chains linked by hydrogen bonds. AB - In the title compound, [Ca(C9H11N6O5)2(H2O)3], the Ca atom lies on a twofold rotation axis in C2/c and the three water molecules are all disordered, each over two sites having equal occupancy. The anion acts as a bridging ligand between pairs of Ca sites on the same twofold axis, thus forming a one-dimensional coordination polymer, with the chains lying along the twofold axes. These chains are linked by multiple O-H...O and N-H...O hydrogen bonds into a single three dimensional framework. PMID- 11408670 TI - Mu-1,4-benzenedicarboxylato-bis[trans-aqua(1,4,8,11 tetraazacyclotetradecane)nickel(II)] diperchlorate forms a three-dimensional hydrogen-bonded framework. AB - The title compound, [Ni2(C8H4O4)(C10H24N4)2(H2O)2](ClO4)2, contains two independent octahedral Ni(II) centres with trans-NiN4O2 chromophores. The bridging benzenedicarboxylate ligand is bonded to the two Ni atoms, each via one O atom of each carboxylate, while the other O atom participates in an intramolecular N-H...O hydrogen bond, forming an S(6) motif. The cations are linked to the perchlorate anions via O-H...O and N-H...O hydrogen bonds [O...O 2.904 (6) and 2.898 (6) A; O-H...O 158 (6) and 165 (6) degrees; N...O 3.175 (7) and 3.116 (7) A; N-H...O 168 and 166 degrees ] to form molecular ladders. These ladders are linked by further O-H...O and N-H...O hydrogen bonds [O...O 2.717 (6) and 2.730 (5) A; O-H...O 170 (4) and 163 (6) degrees; N...O 3.373 (7) and 3.253 (7) A; N-H...O 163 and 167 degrees ] to form a continuous three-dimensional framework. The perchlorate anions both participate in three hydrogen bonds, and both are thus fully ordered. PMID- 11408671 TI - Ferrocenecarboxylic acid-1,4-diazabicyclo[2.2.2]octane (2/1): sheets built from O -H...N and C--H...O hydrogen bonds. AB - In the title compound, 2[Fe(C5H5)(C6H5O2)].C6H12N2, the molecular components are linked into finite three-component aggregates by strong O-H...N hydrogen bonds [O.N 2.578 (4) and 2.604 (5) A; O-H...N 170 (5) and 174 (6) degrees ]; these aggregates are further linked by C-H...O hydrogen bonds [C...O 3.327 (5)-3.401 (5) A; C-H...O 149-157 degrees ] into continuous sheets in the form of (6,3) nets. PMID- 11408672 TI - A new polymorph, form C, of [1,2-bis(diphenylphosphino)ethane] dichloronickel(II). AB - The title compound, [NiCl2(C26H24P2)], has arisen as a result of the unexpected reduction (hydrogenation) of the trans-1,2-bis(diphenylphosphino)ethene ligand. The hydrothermal reaction conditions have produced a third polymorphic form of the compound which has twofold symmetry, crystallizes in an enantiomer-selective manner and contains an unexpectedly short C-C (ethane) bond. Contacts of the form C-H...Cl are present, one involving alkyl and the other aryl hydrogen, with C.Cl distances of 3.556 (4) and 3.664 (6) A, respectively. PMID- 11408673 TI - (12,17-diethoxycarbonyl-2,3,6,7,11,18-hexamethylcorrphycenato kappa4N)iodoiron(III) chloroform solvate. AB - The title complex, (diethyl 3,4,8,15,19,20-hexamethyl-21,22,23,24 tetraazopentacyclo[16.2.1.1(2,5).1(7,11).1(14,17)]tetracosa 1(20),2(22),3,5,7,9,11,13(24),14,16,18-undecaene-9,14-dicarboxylate kappa4N)iodoiron(III) chloroform solvate, [Fe(C32H32N4O4)I].CHCl3, shows an almost planar arrangement of the corrphycene moiety with a slightly distorted trapezoid pyramidal core; the Fe(III) atom is 0.416 (1) A from the plane of the C20N4 system. The Fe-N distances are 2.049 (3), 2.044 (3), 2.079 (3) and 2.075 (3) A. The solvated chloroform forms a C-H...O hydrogen bond [C...O 3.107 (10) A] to an adjacent carbonyl O atom. This is the first X-ray structure analysis of a corrphycenatoiron(III) derivative. PMID- 11408674 TI - A molybdenum complex with 4,7-diphenyl-1,10-phenanthroline. AB - In the title compound, tetracarbonyl(4,7-diphenyl-1,10-phenanthroline N,N')molybdenum(0), [Mo(C24H16N2)(CO)4], the Mo-atom coordination is distorted octahedral, with two CO groups cis to each other, but each trans to an N atom of the 4,7-diphenyl-1,10-phenanthroline (dpphen) ligand, and with the other two CO groups trans to each other and on the axis position. The complex has better solubility than [Mo(phen)(CO)4], where phen is 1,10-phenanthroline. PMID- 11408675 TI - Polynuclear coordination compounds of alkali metal ions with organic chromophores. AB - The crystal structures of sodium 4-([4-[N,N-bis(2 hydroxyethyl)amino]phenyl]diazenyl)benzoate 3.5-hydrate, Na+*C17H18N3O4-*3.5H2O, (I), and potassium 4-([4-[N,N-bis(2-hydroxyethyl)amino]phenyl]diazenyl)benzoate dihydrate, K+*C17H18N3O4-*2H2O, (II), are described. The results indicate an octahedral coordination around sodium in (I) and a trigonal prismatic coordination around potassium in (II). In both cases, coordination around the metal cation is achieved through O atoms of the water molecules and hydroxy groups of the chromophore. The organic conjugated part of the chromophore is approximately planar in (I), while a dihedral angle of 30.7 (2) degrees between the planes of the phenyl rings is observed in (II). PMID- 11408676 TI - Mu-oxo-bis[eta5-1,2-dicarbaundecarboato)bis(N,N-dimethylacetamidinato)-tantalum] dichlooromethane hemisolvate. AB - The structure of the title compound, [Ta2O(C2B9H11)2(C4H9N2)4]*0.5CH2Cl2, contains two (C2B9H11)Ta[NC(Me)NMe2]2 units bridged by a nearly linear [Ta-O-Ta 163.4 (4) degrees ] mu-oxo ligand. The dichloromethane molecule lies on a twofold axis. PMID- 11408677 TI - Aqua(benzene-1,3-dicarboxylato)zinc(II). AB - The title compound, poly[[aquazinc(II)]-mu-benzene-1,3-dicarboxylato-O1:O1':O2], [Zn(C8H4O4)(H2O)]n, forms a metal-organic coordination network that consists of tetrahedral Zn atoms bonded to one water molecule and three carboxylate groups. Isophthalate groups bridge the four-coordinate Zn centers to generate two dimensional architectures in the ac plane. These planar zinc isophthalate motifs are linked by infinite C=O...H-O-H interactions along the a axis to form a chiral framework. The observed polar structural pattern originates due to the distorted tetrahedral Zn centers [O-Zn-O 100.7 (2)-136.0 (1) degrees ] and the alignment of the water molecules. Bridging isophthalate groups align to form approximate centrosymmetric motifs. PMID- 11408678 TI - Bis(2,3-dimethylquinoxalinium) tribromocuprate(I). AB - The title compound, (C10H11N2)2[CuBr3], contains layers of planar monoprotonated cations. Isolated trigonal-planar [CuBr3]2- anions are hydrogen bonded to cations in adjacent layers, providing three-dimensional stability to the crystal structure. PMID- 11408679 TI - Bis(eta2-cyclooctene)(N,N-dimethyldithiocarbamato-S,S')iridium(I), the first example of a tetracoordinate Ir(I) complex containing twopi-bonded monoolefin ligands. AB - The title compound consists of [Ir(C3H6NS2)(C8H14)2] molecules lying on positions with site symmetry 2. Both the coordination plane, defined by the metal, S atoms and the two midpoints of the olefinic bonds, and the dithiocarbamate chelate system are essentially planar. The orientation of the coordinated C=C bonds with respect to the coordination plane is close to perpendicular [(C=C,Ir)/(Ir,S,S) interplanar angle: 79.4 (2) degrees ]. The Ir-C distances are 2.144 (3) and 2.155 (3) A, and the Ir-S bond length is 2.3661 (8) A. Due to pi-coordination, the olefinic bonds are elongated to 1.424 (5) A. The cyclooctene ligands adopt a crown conformation. PMID- 11408680 TI - Bis[tris(3-cyclohexylpyrazol-1-yl)-hydridoborato]copper(II) dichloromethane disolvate. AB - The title compound, [Cu(C27H40BN6)2]*2CH2Cl2, contains a four-coordinate Cu(II) ion lying on a crystallographic inversion centre, giving rise to a near-regular square-planar stereochemistry. There is an axial contact of 2.71 A between the Cu ion and ligand B-H group, although this is unlikely to correspond to a significant 'agostic' interaction. PMID- 11408681 TI - (2-Methylquinolin-8-olato)iron(III) and -copper(II) complexes. AB - The crystal structures of tris(2-methylquinolin-8-olato-N,O)iron(III), [Fe(C10H8NO)3], (I), and aquabis(2-methylquinolin-8-olato-N,O)copper(II), [Cu(C10H8NO)2(H2O)], (II), have been determined. Compound (I) has a distorted octahedral configuration, in which the central Fe atom is coordinated by three N atoms and three O atoms from three 2-methylquinolin-8-olate ligands. The three Fe O bond distances are in the range 1.934 (2)-1.947 (2) A, while the three Fe-N bond distances range from 2.204 (2) to 2.405 (2) A. In compound (II), the central Cu(II) atom and H2O group lie on the crystallographic twofold axis and the coordination geometry of the Cu(II) atom is close to trigonal bipyramidal, with the three O atoms in the basal plane and the two N atoms in apical positions. The Cu-N bond length is 2.018 (5) A. The Cu-O bond length in the basal positions is 1.991 (4) A, while the Cu-O bond length in the apical position is 2.273 (6) A. There is an intermolecular OW-H...O hydrogen bond which links the molecules into a linear chain along the b axis. PMID- 11408682 TI - Bis(O,O'-di-p-tolyldithiophosphato-S,S')(1,10-phenanthroline-N,N')nickel(II). AB - The title coordination complex, [Ni(C14H14O2PS2)2(C12H8N2)] or [Ni(pMePh dtp)2(phen)] (phen is 1,10-phenanthroline; dtp is diaryldithiophosphate), has a non-crystallographic twofold axis of symmetry through the Ni atom and the phen moiety. Two O,O-di-p-tolyldithiophosphate (dtp) ions act as bidentate ligands. The central metal atom is coordinated by four S atoms from two dtp groups and two N atoms from the phen ligand. The title compound displays distorted octahedral geometry around the central Ni atom. PMID- 11408683 TI - A carboxylato-supported alkoxo-bridged dimanganese(III) complex: bis(mu-benzoato O:O')bis[3-(33-methoxysalicydeneamino)propanolato-O,N,O':O']dimanganese(III). AB - The title compound, [Mn2(C11H13NO3)2(C7H5O2)2], is a centrosymmetric dinuclear manganese(III) complex in which the two Mn atoms are bridged by two alkoxo groups and supported by two carboxylate groups, with an Mn.Mn distance of 2.8720 (15) A. PMID- 11408684 TI - Complex of cobalt(II) bromide with hexamethylphosphoric triamide. AB - The title compound, dibromobis[tris(dimethylamino)phosphine oxide]cobalt(II), [CoBr2(C6H18N3OP)2], displays tetrahedral coordination about cobalt. The molecule has twofold crystallographic site symmetry. The short P-N bonds and the planarity of the dimethylamino groups indicate the importance of dpi-ppi interactions. One of the NMe2 groups has an irregular conformation about the P-N bond and deviates from planarity. It is ascribed to the steric hindrance induced by coordination at the O atom of hexamethylphosphoric triamide. PMID- 11408685 TI - 3-amino-4'-methyl-5-ethylbiphenyl-2,4-dicarbonitrile and 3-amino-4'-(n,n diethylamino)-5-ethylbiphenyl-2,4-dicarbonitrile. AB - In the title compounds, C17H15N3 and C20H22N4, the methyl derivative crystallizes with two molecules in the asymmetric unit, while the N,N-diethylamino derivative crystallizes with one molecule per asymmetric unit. The biphenyl twist angle for both molecular structures is approximately 45 degrees. The molecular packing is stabilized by N-H...N hydrogen bonds. PMID- 11408686 TI - 5-amino-4-(4-diethylaminophenyl)-2-(4-hydroxyphenyl)-7-(pyrrolidin-1-yl)-1,6 naphthyridine-8-carbonitrile. AB - In the title compound, C29H30N6O, the naphthyridine moiety is planar with a dihedral angle between the fused rings of 1.9 (1) degrees. The phenol ring is nearly coplanar, while the diethylaminophenyl substituent is orthogonal to the central naphthyridine ring and the pyrrolidine ring makes an angle of 11.2 (1) degrees with it. The O atom of the hydroxy substituent is coplanar with the phenyl ring to which it is attached. The molecular structure is stabilized by a C H...N-type intramolecular hydrogen bond and the packing is stabilized by intermolecular C-H...pi, O-H...N and N-H...O hydrogen bonds. PMID- 11408687 TI - Form II of monoclinic methyl beta-carboline-3-carboxylate. AB - The crystal structure of the second monoclinic P2(1)/c form of the beta-carboline 3-carboxylate, C13H10N2O2, has been determined. Very small changes in the packing scheme lead to a different unit cell; the role of weak C-H...O hydrogen bonds seems to be crucial. PMID- 11408688 TI - Comparison of conventional and synchrotron X-ray structure determinations of the adduct 1,2,4,5-tetrahydroxybenzene-2,5-dihydroxy-1,4-benzoquinone (1/1) and a conventional X-ray structure determination of 1,2,4,5-tetrahydroxybenzene monohydrate. AB - The X-ray structure of 1,2,4,5-tetrahydroxybenzene (benzene-1,2,4,5-tetrol) monohydrate, C6H6O4*H2O, (I), reveals columns of 1,2,4,5-tetrahydroxybenzene parallel to the b axis that are separated by 3.364 (12) and 3.453 (11) A. Molecules in adjacent columns are tilted relative to each other by 27.78 (8) degrees. Water molecules fill the channels between the columns and are involved in hydrogen-bonding interactions with the 1,2,4,5-tetrahydroxybenzene molecules. The crystal structure of the adduct 1,2,4,5-tetrahydroxybenzene-2,5-dihydroxy-1,4 benzoquinone (1/1), C6H6O4*C6H4O4, (II), reveals alternating molecules of 1,2,4,5 tetrahydroxybenzene and 2,5-dihydroxy-1,4-benzoquinone (both lying on inversion centers), and a zigzag hydrogen-bonded network connecting molecules in three dimensions. For compound (II), the conventional X-ray determination, (IIa), is in very good agreement with the synchrotron X-ray determination, (IIb). When differences in data collection temperatures are taken into account, even the displacement parameters are in very good agreement. PMID- 11408689 TI - cis-6-Phenyl-4bH,6H,11H,13H-isoindolo[1,2-c]benz[2,4]oxazepin-13-one. AB - The title compound, C22H17NO2, contains an isoindolinone moiety joined to a phenyl-substituted benzoxazepine ring. The isoindolinone moiety is essentially planar and the oxazepine ring adopts a distorted chair conformation, with the phenyl substituent equatorial. Owing to the severe puckering of the central oxazepine ring, the molecule as a whole is non-planar; the benzene ring of the benzoxazepine fragment makes an angle of 67.7 (1) degrees with respect to the isoindoline ring. PMID- 11408690 TI - 4-(4-Methoxyphenylamino)-3-phenylazo-3-penten-2-one. AB - The title compound, C18H19N3O2, was obtained by an azo-coupling reaction with enaminones and is composed of a planar azoenamine skeleton which forms a six membered ring through a symmetrical intramolecular hydrogen bond. The compound was found to exist as an equilibrium mixture of major hydrazoimino and minor azoenamine tautomers. Quantification of the relative contribution of the tautomeric forms is obscured by the existence of the hydrogen bond. Comparison of the results with those obtained for a similar structure revealed a substantial effect on the tautomeric equilibria of the nature of the substituent bonded to the amine nitrogen. PMID- 11408691 TI - (Z)-7-Chloro-3-[(3-chlorophenyl)-methylidene]-4-p-tosyl-3,4-dihydro-2H-1,4 benzoxazine. AB - In the title compound, C22H17Cl2NO3S, the molecule is a substituted 3,4-dihydro 2H-1,4-benzoxazine compound which has three phenyl rings which are essentially planar. The 3,4-dihydro-2H-oxazine part of the molecule is fused to the benzo ring and has a half-boat conformation; the dihedral angle between the planar part of the oxazine ring and the benzo ring is 10.2 (2) degrees. The (3 chlorophenyl)methylidene substituent has a Z configuration in relation to the ring N atom of the oxazine moiety. Interestingly, the p-toluenesulfonyl (p-tosyl) substituent on the ring N atom protrudes away from the 3-chlorophenyl substituent thus avoiding any steric interaction. PMID- 11408692 TI - Solvated and solvent-free forms of N,N'-dithiodiphthalimide. AB - N,N'-Dithiodiphthalimide, C16H8N2O4S2, crystallizes from ethyl acetate with two independent molecules in the asymmetric unit, in which the N-S-S-N torsion angles are -83.59 (19) and 92.9 (2) degrees. The molecules are linked by C-H...O hydrogen bonds and aromatic pi-pi-stacking interactions into a three-dimensional framework. When crystallized from either dichloromethane or ethanol, solvates are formed in which the molecules of the title compound lie across twofold rotation axes in space group C2/c, with N-S-S-N torsion angles of 93.54 (7) and 96.14 (11) degrees. There are no hydrogen bonds in these solvates, but the molecules are linked by aromatic pi-pi-stacking interactions into chains, between which there are continuous channels. Disordered solvent molecules occupy these channels, which account for ca 20% of the unit-cell volume. PMID- 11408693 TI - meso-N,N'-oxalyldivaline. AB - The title compound, 2,2'-(oxalyldiimino)bis(3-methylbutanoic acid), C12H20N2O6, possesses a centre of symmetry. In the crystal, molecules are connected by hydrogen bonds between oxamide and carboxyl groups, similar to the pattern of the monoclinic forms of HO-Gly-CO-CO-Gly-OH and HO-Aib-CO-CO-Aib-OH (Gly is glycine and Aib is 2-aminoisobutyric acid). The characteristic torsion angles in the title compound are close to those in peptide alpha-helices. PMID- 11408694 TI - Acetonyldichloro[(Z)-2-chloro-2-phenylvinyl]tellurium(IV), helical chains of metal complexes. AB - The primary geometry about the Te(IV) atom in the title compound, [TeCl2(C8H6Cl)(C3H5O)] or C11H11Cl3OTe, is a pseudo-trigonal-bipyramidal arrangement, with two Cl atoms in apical positions, and the lone pair of electrons and C atoms in the equatorial plane. The Te(IV) atom is involved in three secondary interactions, two intramolecular [Te...O = 2.842 (3) A and Te.Cl3 = 3.209 (1) A] and one intermolecular [Te...Cl = 3.637 (1) A], the latter giving rise to a helical chain. These helices are linked by C-H...O interchain interactions. PMID- 11408695 TI - 4,7-bis(4-pyridylethynyl)-2,1,3-benzothiadiazole and its dipyridinium diperchlorate. AB - The title compound, C20H10N4S, and its dipyridinium salt, 4,4'-(2,1,3-benzodiazol 4,7-diyldiethynyl)dipyridinium diperchlorate, C20H12N4S2+*2ClO4-, display bond alternation in the 2,1,3-benzothiadiazole rings, which suggests their quinonoid character. The dipyridinium dication molecules stack along the a axis and form a dimer with short S.N interheteroatom contacts [3.146 (4) A] between the two 1,2,5 thiadiazole rings. The dimer is surrounded by the perchlorate anions with which it forms a large number of intermolecular N-H...O and C-H...O hydrogen bonds. PMID- 11408696 TI - 1-methyl-4-nitraminopyridinium nitrate and 4-nitraminopyridinium methanesulfonate. AB - In the title compounds, C6H8N3O2+*NO3- and C5H6N3O2+*-CH3SO3-, respectively, the cations are almost planar; the twist of the nitramino group about the C-N and N-N bonds does not exceed 10 degrees. The deviations from coplanarity are accounted for by intermolecular N-H...O interactions. The coplanarity of the NHNO2 group and the phenyl ring leads to the deformation of the nitramino group. The C-N-N angle and one C-C-N angle at the junction of the phenyl ring and the nitramino group are increased from 120 degrees by ca 6 degrees, whereas the other junction C-C-N angle is decreased by ca 5 degrees. Within the nitro group, the O-N-O angle is increased by ca 5 degrees and one O-N-N angle is decreased by ca 5 degrees, whereas the other O-N-N angle remains almost unchanged. The cations are connected to the anions by relatively strong N-H...O hydrogen bonds [shortest H.O separations 1.77 (2)-1.81 (3) A] and much weaker C-H...O hydrogen bonds [H...O separations 2.30 (2)-2.63 (3) A]. PMID- 11408697 TI - (E)-1-(2'-deoxy-beta-D-ribofuranosyl)-2,4-difluoro-5-(2-iodovinyl)benzene. AB - This analysis of the title compound, C13H13F2IO3, establishes the orientation of (E)-5-(CH=CH-I) as antiperiplanar (ap) to the C-C bond (5-6 position) of the 2,4 difluorophenyl ring system, with the (E)-5-(CH=CH-I) H atom located in close proximity (2.17 A) to the F4 atom of the 2,4-difluorophenyl moiety. PMID- 11408698 TI - Betainium trifluoroacetate. AB - The title compound, C5H12NO2+*C2F3O2- or BET+*CF3COO- [BET is trimethylglycine (betaine); IUPAC: 1-carboxy-N,N,N-trimethylmethanaminium inner salt], contains pairs of betainium and trifluoroacetate ions forming a dimer bridged by a strong hydrogen bond between the carboxyl and carboxylate groups of the two ions. The molecular symmetry of the cation is close to Cs, with protonation occurring at the carboxy O atom positioned anti to the N atom. The trifluoroacetate anions are disordered over two positions. In one, the conformation of the CF3 group is staggered with respect to the carboxylate group, in the other, it is close to an eclipsed conformation. The sole hydrogen bond present in the structure is the strong O-H...O bond between the anion and the cation. PMID- 11408699 TI - Dispiro[fluorene-9,5'-[1,2,3,4]tetrathiane-6',9"-fluorene. AB - The tetrathiane ring of the title compound, C26H16S4, has a chair conformation and the molecule has approximate C2 symmetry. Each of the two fluorene ring systems is virtually planar, with the ring planes intersecting at an angle of 67.58 (5) degrees. This novel compound has been formed as a side product from the treatment of 9H-fluorene-9-thione with methyl N-[(benzylidene)phenyl]glycinate in the presence of LiBr and 1,6-diazabicyclo[5.4.0]undecane. PMID- 11408700 TI - Water molecules in the crystal structure of tricyclic acyclovir. AB - The biologically important molecule tricyclic acyclovir, presented here as 3-[(2 hydroxyethoxy)methyl]-6-methyl-3H-imidazolo[1,2-a]purin-9(5H)-one dihydrate, C11H13N5O3*2H2O, shows conformational flexibility, which is observed in the solid state as two symmetrically independent molecules with different side-chain conformations. Additionally, one of these molecules exhibits side-chain disorder, such that there are three different conformations in the crystal. Water molecules found in the crystal form (H2O)8 clusters which are located between molecules of tricyclic acyclovir. The complex hydrogen-bond network formed between water and tricyclic acyclovir in the solid state may be related to the solvation of the molecules in solution. PMID- 11408701 TI - Bis(mu-pyrrolidine-N:N)bis(dibromoboron), [(Br)2B(pyrrolidine)2B(Br)2]. AB - The B atoms in the title compound, C8H16B2Br4N2, bridge between the two monomeric moieties, forming a (BN)2 four-membered ring with partial bond orders of the B-N bonds. PMID- 11408702 TI - Dimorphism of ethyl 1,4-dihydro-2,4,6-triphenylpyridine-3-carboxylate. AB - The title compound, C26H23NO2, (Ia) and (Ib), shows polymorphism with crystals obtained from different solvents displaying different crystal structures. However, it is not the geometry of the single molecules nor the hydrogen-bond pattern that is different in (Ia) and (Ib), but the way in which the hydrogen bonded chains, running along the a-axis direction, are arranged with respect to each other. PMID- 11408703 TI - The C-H...O hydrogen bond in (dicyanomethyl)ammonium p-toluenesulfonate. AB - In the title compound, C3H4N3+*C7H7O3S-, the activated C-H group of the cation forms a short but bent C-H.O hydrogen bond with a sulfonate O atom of the anion; C.O = 3.075 (5) A and C-H...O = 130 degrees. PMID- 11408704 TI - N-(4-Biphenylyl)urea. AB - In the crystal structure of the title compound, C13H12N2O, N-H(anti)...O hydrogen bonds produce the so-called urea alpha-network and the N-H(syn) donor forms an unconventional N-H...pi hydrogen bond. PMID- 11408705 TI - Perivascular plasmacytosis: a light-microscopic and immunohistochemical study of 93 bone marrow biopsies. AB - In bone marrow biopsies from 13 hematologically normal persons and from 80 patients with a variety of disorders, we found perivascular plasmacytosis. In all instances except the one case of multiple myeloma, plasma cells were polyclonal and normal in morphology. This was especially pronounced in patients with HIV infection, and in individuals following chemotherapy. In the same patients, sinusoids were also prominent, and appeared dilated. In biopsy sections, small endothelial-lined vessels appeared to arise from attenuation of the sinusoidal lumen. After a short segment of only endothelial-lined vessels, perivascular plasmacytosis appeared. When smooth muscle cells began to line the endothelium, plasma cells virtually disappeared. The biological significance of this finding is unknown. Possibly, the close proximity of plasma cells to endothelial cells early in the development of blood vessels could facilitate entry of immunoglobulins into the blood. PMID- 11408706 TI - Remarkably reduced transplant-related complications by dibromomannitol non myeloablative conditioning before allogeneic bone marrow transplantation in chronic myeloid leukemia. AB - A non-myeloablative conditioning protocol containing dibromomannitol (DBM/cytosine arabinoside/cyclophosphamide) has been applied to 36 chronic myeloid leukemia (CML) patients followed by bone marrow transplantation (BMT) from sibling donors. Risk factors include: accelerated phase (10 patients), older age (17 patients over >40 years) and long interval between diagnosis and BMT (27 months on average). Severe mucositis did not occur. Venoocclusive liver disease was absent. Infectious complications were rare. Although grade II-IV acute graft versus-host disease (GVHD) was present in 9 (25%) cases, there were only 2 serious (III-IV) ones. Chronic GVHD occurred in 25 (69%) cases, preceded by acute GVHD in 9 of the 25 affected patients. Early hematological relapse, 7-29 weeks after BMT, developed in 6 patients (17.6%). No relapse was noted in the completely chimeric patients, however molecular genetic residual disease was observed in 6 patients, in most of them after transient short-term mixed chimeric state. Overall actual survival rate is 83.3% for the 36 cases, and leukemia-free survival is 72.2% for the 34 engrafted patients. PMID- 11408707 TI - Hematologic and immunophenotypic characterization of human umbilical cord blood. AB - In this work, cord blood cells from 30 healthy term newborns were analyzed for complete blood counts with an automated cytometer and, in part of the sample, for surface molecules in cord blood monocytes, lymphocytes and CD34+ cells by two color flow cytometry. Hematological parameters were as follows: WBC = 12.85 (5.24 15.10) x10(9)/l; platelets = 304.33 (156.00-469.00) x 10(9)/l; Hb = 14.45 (11.90 17.82) g/dl; RBC = 3.99 (3.14-5.12) x 10(12)/l; MCV 107.25 (99.60-115.00) fl; reticulocytes = 157.80 (101.00-124.00) x 10(9)/l or 3.99 (2.45-6.01)%; erythroblasts = 0.88 (0.15-2.58) x 10(9)/l or 6.63 (2.86-16.80) per 100 WBC [corrected]. The mononuclear population, as evaluated by flow cytometry, was composed of 22.9 +/- 7.2% monocytes and 77.05 +/- 7.24% lymphocytes, among which 46.59 +/- 15.62% were T lymphocytes (43.94 +/- 16.94% CD3+/CD4+ and 13.45 +/- 7.46% CD3+/CD8+). CD34+ cells were on average 0.54 +/- 0.24% of the mononuclear fraction. CD11c, CD49e and HLA-DR were found mainly on monocytes, and CD31 and CD62L occurred in similar levels on monocytes and lymphocytes. CD117+ cells were less than 5% of these populations. Among CD34+ cells, CD31 and HLA-DR were the molecules with higher frequencies (79.7 +/- 19.9 and 65.7 +/- 23.0%, respectively), followed by CD62L (41.8 +/- 31.9%) and CD117 (20.1 +/- 15.8%). The presence of CD11c and CD49e on CD34+ cells was low (below 10%). The results stress the phenotypic heterogeneity of cord blood CD34+ cells, and the different behavior of the cells when manipulated in vitro in different degrees of isolation. PMID- 11408708 TI - Activity of a nitroxylated analog of daunorubicin, ruboxyl, in B lymphoproliferative disorders. AB - A nitroxylated analog of daunorubicin, ruboxyl (RBX), showed low toxicity but significant lympholytic effect in preclinical evaluations. A series of studies in vitro and in animals demonstrate that RBX is a putative agent in the treatment of many neoplasms. We report the results of a study in mice in which RBX showed selective B-lymphocyte immunosuppression. On the basis of this experience, RBX was administered to 3 patients with multiple myeloma and two patients with Waldenstrom's disease. The results of this pilot clinical study show that this compound has good activity and low myelotoxicity and cardiotoxicity, but seems to be characterized by a threatening immunosuppressive effect. PMID- 11408709 TI - Mucosa-associated lymphoid tissue lymphoma of the salivary glands occurring in patients affected by Sjogren's syndrome: report of 6 cases. AB - OBJECTIVE: Mucosa-associated lymphoid tissue (MALT) lymphoma of the salivary glands occurring in 6 patients affected by primary Sjogren's syndrome is reported. METHODS: Clinical findings, histologic type, stage, treatment and outcome of the 6 patients have been revised. RESULTS: In all 6 cases the lymphoma was of the MALT type. Four patients had stage IE disease, 1 patient had stage IIE disease and 1 patient had stage IV disease. The patients received different treatments resulting in all cases in prolonged remission. After 7 years of complete remission 1 patient developed a diffuse large B-cell lymphoma. CONCLUSION: MALT lymphoma of the salivary glands is an indolent disease. Though the best therapy of this lymphoproliferative disorder remains to be established, prolonged remission has been obtained in our cases with different therapeutic approaches. We review the literature regarding the relationship between Sjogren's syndrome and MALT lymphomas and study the mechanisms which may be involved in the transformation from a lymphoepithelial lesion into a neoplastic disorder. PMID- 11408710 TI - Increased proportion of HLA-class-I-specific natural killer cell receptors (CD94) on peripheral blood mononuclear cells after allogeneic bone marrow transplantation. AB - In the present study, we investigated the inhibitory natural killer cell receptor (NKR) expression of CD94/NKG2A on PBMC after allogeneic bone marrow transplantation (BMT). The proportion of CD94 expression on PBMC was higher in patients without chronic graft-versus-host disease (cGVHD) and also in cGVHD patients with good response to conventional immunosuppressive therapy than in cGVHD patients with poor response. Also, the proportions of CD94+/CD3+ cells and CD94+/CD8+ cells were higher in cGVHD patients showing good response. In addition, the proportion of NKG2A-expressing cells was higher in patients without cGVHD than in patients with cGVHD. Therefore, chronic allostimulation after allo BMT may augment the proportion of CD94/NKG2A-positive cells, and these cells may play some role in the regulation of alloresponse in some patients. PMID- 11408711 TI - Primary lung involvement in Waldenstrom's macroglobulinaemia: report of two cases and review of the literature. AB - Pulmonary involvement in Waldenstrom's macroglobulinaemia (WM) occurs in 3-5% of cases, but lung involvement without bone marrow infiltration is extremely rare. We report 2 patients who presented with bilateral consolidations on chest X-ray and non-specific symptoms and were treated for a long period of time for pulmonary infections until the diagnosis was made by open lung biopsy. Both patients presented high monoclonal IgM in the serum and one also had blood lymphoplasmacytosis. Trephine bone biopsy and bone marrow smears were normal and there was no other site of involvement. Along with the presentation of our patients, we review the literature, discuss some of the possible underlying mechanisms and raise the attention of clinicians to this rare manifestation of the disease. PMID- 11408712 TI - Increased warfarin sensitivity as an early manifestation of occult prostate cancer with chronic disseminated intravascular coagulation. AB - Increased sensitivity to warfarin anticoagulation is usually attributed to liver disease, vitamin K deficiency, or drug interactions. We describe a patient with unexplained sensitivity to warfarin and mildly elevated prostate-specific antigen levels in whom subsequent developments indicated that warfarin sensitivity was the first manifestation of occult prostatic cancer. A review of all published cases of coagulopathy associated with cancer of the prostate shows that, unlike other solid tumors with secondary disseminated intravascular coagulation (DIC), in prostate cancer increased bleeding is more common than thrombotic phenomena. Chronic DIC due to occult prostate cancer should be included in the differential diagnosis of excessive prothrombin time prolongation in patients receiving anticoagulants. PMID- 11408713 TI - Myelodysplasia-like syndrome and end-stage liver disease: report of 2 cases. AB - We report 2 cases of myelodysplastic syndrome associated with cirrhosis caused by primary biliary cirrhosis and alpha1-antitrypsin deficiency. The etiology of myelodysplasia and its implication in liver transplantation is discussed. PMID- 11408714 TI - Adult T-cell leukemia with anti-HTLV-I antibody but no HTLV-I DNA in tumor cells. AB - Adult T-cell leukemia (ATL) is usually defined as a malignant disease of T cells infected by human T-lymphotropic virus type I (HTLV-I). In the present study, we describe a 49-year-old woman with an acute type ATL, whose leukemic cells do not contain the HTLV-I genome. Laboratory tests revealed an increase in abnormal lymphocytes with convoluted nuclei, elevated serum lactate dehydrogenase levels, increased thymidine kinase activity and soluble interleukin-2 receptor-alpha levels. Serum examination demonstrated positive anti-HTLV-I antibody, but Southern blot analysis using the whole HTLV-I genome as a probe did not detect any integration of the viral genome. In contrast, PCR detected the HTLV-I pX region in the same DNA samples as used for Southern blot analysis. These findings suggest two possibilities. One possibility is that ATL in this patient is generated by other pathogens than HTLV-I virus. She is also an HTLV-I carrier. The other possibility is that her leukemic T cell clone derived its malignant phenotype from HTLV-I infection, and once this malignant phenotype was obtained, partial deletions of viral genome repeated until the whole viral genome was deleted. Although there is no direct evidence, the former possibility is more likely in the present case. PMID- 11408715 TI - Successful treatment of angioimmunoblastic lymphadenopathy with dysproteinemia type T-cell lymphoma with fludarabine. PMID- 11408716 TI - Plasma chitotriosidase activity is a marker of recovery in transplanted patients affected by beta-thalassemia major. PMID- 11408717 TI - VLA-4 expression and cell cycling status during ex vivo expansion of human cord blood CD34+ cells. PMID- 11408718 TI - Successful treatment of granulocytic sarcoma with alpha-interferon and disodium pamidronate at presentation of chronic myeloid leukemia. PMID- 11408719 TI - Megadose methylprednisolone for granulocytic sarcoma. PMID- 11408720 TI - Tuberculosis in the elderly. AB - Tuberculosis is still a major cause of morbidity and mortality worldwide. Recent studies have suggested that even in the developed world its incidence in the elderly is increasing. Symptoms of active tuberculosis are non-specific and less pronounced in the elderly. Radiological features are more likely to be 'atypical' in the older subject and skin tests more frequently negative. This results in delay in diagnosis and higher mortality from tuberculosis in the aged population. A high degree of clinical suspicion is therefore required to ensure the diagnosis is not missed and appropriate treatment instigated. PMID- 11408721 TI - Age-related alterations in basal expression and in vitro, tumour necrosis factor alpha mediated, upregulation of CD11b. AB - BACKGROUND: The beta(2-)integrin CD11b (Mac-1) plays a crucial role in the firm attachment of leucocytes to the endothelium during the inflammatory response. OBJECTIVE: This study aimed to determine whether the increased incidence of infections witnessed in elderly individuals compared to their younger counterparts was associated with deficiencies in basal expression and/or upregulation of CD11b. METHODS: Flow cytometry was used to measure CD11b expression, before and after in vitro tumour necrosis factor alpha (TNF-alpha) stimulation, on neutrophils, monocytes and lymphocytes from healthy volunteers aged less than 36 years and Senieur-approximated 70-85 and over 85 year olds. The TNF-alpha levels in serum were measured using a commercially available enzyme linked immunoassay technique. RESULTS: The basal expression of CD11b on monocytes and lymphocytes was highest in the 70-85-year-olds and lowest in the > 85-year olds. Following in vitro stimulation using low (10 IU) and high (100 IU) TNF alpha concentrations, subjects > 85 years consistently showed significantly lower increases in CD11b expression on each of the three cell types. The maximal increase in CD11b expression was in the 70-85-year age group for neutrophils and monocytes and in < 36-year-olds for lymphocytes. Serum TNF-alpha was significantly higher in the elderly groups. Regression analysis showed a significant association between TNF-alpha and expression of CD11b on lymphocytes before and after TNF-alpha stimulation and for neutrophils before stimulation. CONCLUSIONS: The results of this study suggest that CD11b expression on leucocytes may not be consistent throughout life. Such age-related changes could compromise the inflammatory response, rendering individuals > 85 years old more susceptible to infections. Alternatively, the lower levels of CD11b expression in this group may represent downregulation and protection against excess leucocyte activation within the vascular system and may, therefore, provide a mechanism for successful ageing. PMID- 11408722 TI - Antioxidant enzyme levels in red blood cells from cataract patients. AB - BACKGROUND: It seems very likely that oxidative mechanisms play a major role in aetiology and pathogenesis of senile cataract. In particular, lens proteins are subject to extensive oxidative modifications. OBJECTIVE: The purpose of this work was to analyze the activities of the protective enzymes superoxide dismutase (SOD) and catalase (CAT) in patients of both sexes affected by cataract. METHODS: The SOD activity was measured in red blood cells using the Minami and Yoshikawa method, and the CAT activity was measured in haemolysates by the method of Aebi. The results were compared with those obtained in a group of healthy subjects of both sexes and matched ages. RESULTS: The SOD activity shows a significant increase when compared with controls, whereas the CAT activity was not modified. CONCLUSIONS: The balance of the anti-oxidants in red blood cells from cataract patients is altered. PMID- 11408723 TI - Age-related changes of postural control: effect of cognitive tasks. AB - BACKGROUND: Postural control and falls in the elderly constitute a major health problem. The interest in balance deficits is growing, as concern about the rising costs of health care increases. This issue is particularly relevant to the elderly population in which falls occur most frequently. Postural control in the elderly was studied using a cognitive approach. OBJECTIVE: The purpose of this study was to study the characteristics of central processing of postural control while performing cognitive tasks. METHODS: A dual-task procedure was developed to estimate the level of automaticity of a quiet upright standing task. The effect of a concurrent attention-demanding task (modified Stroop test) on the efficiency of balance control in the elderly was determined using force platform and electromyography measurements. RESULTS: It was found that there is an increase in postural sway in old subjects compared with young subjects when performing single tasks and dual-task tests. The results of the study demonstrate that postural adjustments require cognitive processing; young and old subjects showed similar interference effects on postural steadiness (postural sway) caused by the concurrent attention-demanding task. The results are corroborated by the hypothesis that a dual task gives information on the restoration of automaticity of postural control in old age by a central reorganization process. When performing a dual task tested on a narrow base of support, the old subjects decreased their body sway, while the younger did not. According to electromyography measurements, the older subjects increased their muscle activity in the tibialis anterior and soleus muscles, using slow-twitch motor units compared with the younger subjects. CONCLUSIONS: Both alterations (cognitive and base of support) have a substantially greater effect on the elderly than on the young. The older subjects decreased their body sway by activating a cocontraction strategy of postural control around the ankle joint, probably because of the danger to their postural stability. PMID- 11408724 TI - Suspected cardiac syncope in elderly patients: use of the 12-lead electrocardiogram to select patients for holter monitoring. AB - BACKGROUND: Results of 24-hour Holter monitoring in elderly patients are often unhelpful, since the prevalence of asymptomatic arrhythmias increases and their prognostic significance is unclear. We investigated the value of the resting electrocardiogram (ECG) in predicting significant findings on 24-hour Holter recordings in those suspected of having cardiac syncope. OBJECTIVE: To see whether the resting 12-lead ECG can be used as a screening tool to select elderly patients suspected of having cardiac syncope for 24-hour ECG monitoring. METHOD: Comparison of resting 12-lead ECGs and 24-hour Holter tapes in 145 consecutive elderly outpatients suspected of having a cardiac cause for falls, dizziness, or syncope. RESULTS: Four of 30 normal ECGs (13%) showed an abnormality on Holter monitoring as compared with 55 of the 115 abnormal ECGs (47.8%; chi = 11.7143, p < 0.005). In the 'normal' group the 4 abnormal Holter recordings all showed short runs of supraventricular tachycardia, and no intervention resulted. The 115 abnormal resting ECGs showed either ischaemia (n = 27), dysrhythmia (n = 28), sinus bradycardia (n = 22), or conduction defects (n = 38). The 55 of these which showed abnormalities on Holter recordings occurred mostly where the resting ECG showed dysrhythmia (n = 14/28; 50%), bradycardia (n = 19/22; 86.4%), and conduction defect (n = 17/38; 44.7%). Seven patients had complete heart block on Holter, and all had conduction defects on resting ECG (p < 0.0004). Fifteen patients had pauses of longer than 3 s on Holter; all had conduction defect, bradycardia, or dysrhythmias on resting ECG (p < 0.0045). Sixteen patients were paced because of complete heart block or pauses on Holter recordings, and all had either bradycardia or conduction defects on resting ECG, resulting in complete resolution of their symptoms. CONCLUSIONS: Patients with suspected cardiac syncope and normal resting ECGs are unlikely to reveal significant abnormalities on single 24-hour Holter monitoring. Cardiac event recorder or prolonged Holter monitoring may be required in patients with strong clinical history. Those with abnormal ECGs, in particular sinus bradycardia and conduction defects, are highly likely to have significant abnormalities on 24-hour ECG monitoring. PMID- 11408725 TI - Plasma insulin-like growth factor I levels in the elderly: relation to plasma dehydroepiandrosterone sulfate levels, nutritional status, health and mortality. AB - BACKGROUND: Insulin-like growth factor I (IGF-I) has beneficial metabolic effects. Because plasma IGF-I levels have been reported to be enhanced by dehydroepiandrosterone (DHEA) administration, it has been suggested that the IGF I may be implicated in some of the reported associations between low DHEA sulfate (DHEAS) levels and impaired health measures in elderly subjects. The nutritional status, which also regulates plasma IGF-I levels, is also an important determinant of health outcome. OBJECTIVE: We sought to investigate the associations between plasma IGF-I levels and plasma DHEAS levels, nutritional status, health, and mortality in the elderly. METHODS: In 256 community-dwelling subjects aged 65-101 years, enrolled in the Paquid study, a health questionnaire was used to acertain their functional disabilities, any history of medical disorders, self-perceived health, depressive symptoms, and their cognitive function. Biological measurements included levels of plasma IGF-I, albumin, transthyretin, and DHEAS. Mortality data were available for 6 years following blood sampling. RESULTS: In this elderly population with no impairment in nutritional status, the plasma IGF-I levels decreased significantly with age (p = 0.02). The plasma IGF-I levels correlated positively with those of transthyretin (p = 0.0001). IGF-I also correlated with DHEAS (p = 0.04), but the correlation did not remain significant after adjustment for age. As opposed to SDHEA, the plasma IGF-I levels did not correlate with the results from the health questionnaire. The baseline IGF-I values in the highest tertile were associated with a higher risk of short-term mortality than those in the lowest tertile (RR = 8.4 at 2 years, p = 0.007). CONCLUSIONS: Our results suggest that IGF-I is not connected with the association between low plasma DHEAS levels and the impaired results from the health questionnaire. The relationship between plasma IGF-I highest levels and mortality should be further explored. PMID- 11408726 TI - Influence of muscle strength and body weight and composition on regional bone mineral density in healthy women aged 60 years and over. AB - Although weight, lean mass, fat mass and muscular strength are often found to be intercorrelated, the respective role of each parameter in bone mineral density (BMD) remains unknown in older women. The aim of the present study was to investigate the relationship between body weight and composition and quadriceps strength on femoral neck and lumbar spine BMD in healthy postmenopausal women. The relationship between isokinetic quadriceps strength measured by Biodex and BMD measured by dual energy X-ray absorptiometry was studied in 56 women aged 60 81 (70.5 +/- 6.2) years in multiple regression models adjusted for age, body composition and menopausal treatment. Weight and age were associated with femoral neck BMD (33 and 10% of variance accounted for, respectively) and lumbar spine BMD (23 and 8% of its variance). When body weight and quadriceps strength were excluded from the model, lean mass and age were associated with femoral neck BMD (29 and 14% of variance explained, respectively) and lumbar spine BMD (28 and 11% of variance explained, respectively). When quadriceps strength was entered into the model, it was strongly associated with femoral neck BMD (30% of variance accounted for), in addition to lean mass (9%) and age (7%), whereas it was not associated with lumbar spine BMD. In conclusion, lean mass explains a great part of the strong association between body weight and femoral neck and lumbar spine BMD. Quadriceps strength explains a great part of the association between lean mass and BMD at the femoral neck site but not at the lumbar spine site. These results suggest a site-specific effect of muscular strength on bone and a potential role of the age-related decline of muscle strength in age-related bone loss in postmenopausal women. PMID- 11408727 TI - The relationship between apolipoprotein E4 and lipid metabolism is impaired in Alzheimer's disease. AB - BACKGROUND: Human apolipoprotein (Apo) E4 (ApoE4) is an important determinant of lipid metabolism and cell-to-cell cholesterol transport. It is also a major genetic risk factor for both vascular disease and familial and sporadic late onset Alzheimer's disease (AD). Since vascular pathology could dramatically reduce neuronal reserve capacity, a biological chain of events between ApoE4, hypercholesterolemia and AD has been postulated. The aim of the present study is to explore the relationship between lipid metabolism and ApoE isoforms in a large series of elderly subjects in relation to the presence or absence of AD. METHODS: Of 332 referrals to a neurology clinic specializing in memory disorders, 146 were given a diagnosis of AD (age, mean +/- SD 76 +/- 13.1 years, 64.4% women) according to DSM-IIIR criteria. One hundred and seventy-six subjects were included as controls (age 80 +/- 5.6 years, 58% women). The ApoE phenotype was determined by the isoelectrofocalization method, cholesterol, triglycerides, phospholipids, ApoA and ApoB were determined by routine chemistry. FINDINGS: A significant association was observed between the E4 allele and AD (chi2 = 13, p < 0.001) only below age 80. In the control group, cholesterol levels were found to be significantly higher in men but not in women with an E4 allele (6.35 +/- 1.3 mmol/l) as opposed to those without (5.8 +/- 1.2 mmol/l). ApoB levels were also found to be higher in the presence of ApoE4, with no gender effect. Within the AD group no significant relationship was found between ApoE4 and cholesterol levels (mean 6.05 +/- 0.9 mmol/l in E4-AD subjects versus 5.8 +/- 1.21 mmol/l in non-E4 AD subjects). Similar observations were made in relation to triglycerides and phospholipids. INTERPRETATION: Our results demonstrate the disappearance of the ApoE4-raising effect on serum cholesterol, triglyceride and phospholipid levels in AD suggesting a more complex relationship between AD and lipid metabolism than has previously been supposed. This lipid abnormality may further contribute to the progression of AD. PMID- 11408728 TI - Changing colour preferences with ageing: a comparative study on younger and older native Germans aged 19-90 years. AB - BACKGROUND: Previous studies have shown that colour preferences change from early childhood to adulthood. OBJECTIVE: The main objective of this study was to assess whether colour preferences undergo further changes during adult life. METHODS: Cross-sectional data on colour preferences were collected in 842 adults (aged 19 90 years) living in Mainz, Germany. The sample was split by age into younger (183 men, 232 women) and older (157 men, 270 women) adults. The subgroups were compared for their most and least preferred colours that they had chosen by name out of four given colours (blue, green, red, yellow). Differences in the pattern of frequencies between sexes and age groups were assessed for statistical significance using chi-square analyses. RESULTS: Both age groups and both sexes chose blue as most preferred colour and yellow as least preferred one; green and red fluctuated in the middle position of the rank order of preferences. All frequency distributions departed significantly from chance (p < 0.001). In both younger and older adults, significant sex differences did not exist for the ranking of the most preferred colours (p > 0.2), but were evident in the ranking of the least preferred colours (p < 0.01); men stated more often yellow and less often red as least preferred than women did. Age group differences in colour preferences were highly significant. With advancing age, the preference for blue decreased steadily, whereas the popularity of green and red increased (p < 0.001). CONCLUSIONS: The results suggest that colour preferences change during the course of the adult life span. These changes seem to be a reversal of the trend reported in the literature for children. Changed colour preferences in the aged might be attributed to alterations in colour discrimination and visual imagery, the yellowing of the crystalline lens, and the decreased function of the blue cone mechanism with ageing. PMID- 11408730 TI - Involvement of phenotypes and serum levels of haptoglobin in the outcome of in vitro fertilization and embryo transfer. AB - Haptoglobin (Hp) possesses many functions in the immune system and angiogenesis as well as scavenging released hemoglobin, and thus it is speculated that Hp may also play a critical role in the reproductive process. To investigate the possible involvement of Hp in the outcome of in vitro fertilization and embryo transfer (IVF-ET), the serum Hp of 50 patients who underwent IVF-ET was analyzed by the Western blot method. The results showed a higher pregnancy rate in the patients with the Hp2-2 phenotype (38%) than in those with the Hp2-1 phenotype (16%) (p = 0.08), and the serum alpha 2 subunit levels in the patients who achieved pregnancy within four attempts at IVF-ET were higher than in the others (p < 0.05). The results of this study suggest that Hp is involved in the outcome of IVF-ET. PMID- 11408731 TI - Comparison of cerebral blood flow velocity as measured in preeclamptic, healthy pregnant, and nonpregnant women by transcranial Doppler sonography. AB - AIM: To test the hypothesis that the middle cerebral artery blood flow velocity (MCAV) is altered in preeclamptic pregnant women as compared with healthy pregnant and nonpregnant women. METHODS: Preeclamptic (n = 21) and healthy pregnant (n = 17) as well as healthy nonpregnant (n = 29) women underwent transcranial Doppler MCAV measurements. The mean MCAV values were compared between the different groups. Anova combined with Bonferroni correction was used for statistical analysis. RESULTS: The MCAV was significantly higher in nonpregnant women (mean +/- SE 73.0 +/- 2.12 cm/s) as compared with healthy pregnant women (67.0 +/- 1.8 cm/s, p = 0.0356). Preeclamptic women showed significantly higher MCAV values (83.5 +/- 2.1 cm/s) as compared with nonpregnant females (73.0 +/- 2.12 cm/s, p = 0.0014). Similar to nonpregnant women, healthy pregnant women showed lower MCAV values (67.0 +/- 1.8 cm/s) as compared with preeclamptic women (83.5 +/- 2.1 cm/s, p = 0.001). After Bonferroni correction the MCAV values in patients suffering from preeclampsia were still statistically significantly higher as compared with the two other groups. CONCLUSIONS: We detected increased resting MCAV values in pregnant women with preeclampsia. In our opinion, this finding refers to arteriolar dilation of the resistance vessels of the brain. Further studies are needed to prove altered vasoreactivity responses of the brain resistance arterioles in preeclampsia. PMID- 11408732 TI - Expression of nm23-H1 in hydatidiform mole and its relationship with the development of postmolar disease. AB - The aim of the present study was to investigate the expression of nm23-H1 in human placenta, hydatidiform mole and choriocarcinoma cells. Nm23-H1 protein was localized in the cytotrophoblast, but not in the syncytiotrophoblast. In the hydatidiform mole cases with subsequent spontaneous remission, nm23-H1 mRNA levels were significantly lower than those in first-trimester placentas. However, its levels were elevated in the hydatidiform mole cases that progressed to persistent gestational trophoblastic disease and were comparable to those of first-trimester placentas, and they were further elevated in choriocarcinoma cells. The present data suggest an association of nm23-H1 for the proliferation activity of trophoblast, and its increased expression may influence the development of persistent trophoblastic disease. PMID- 11408733 TI - Detection of aberrations of chromosome 17 and p53 gene expression and their correlation to histologic grading and prognosis in primary invasive squamous cell carcinoma of the cervix. AB - We analyzed tumor tissues from 14 patients with invasive squamous cell carcinoma of the cervix for aberrations of chromosome 17 and p53 expression. All but 3 patients were negative for p53 protein expression, the protein being detected in 2 International Federation of Obstetrics and Gynecology stage IIa cancers and 1 stage Ib G3 carcinoma. Significant cytogenetic aberrations in the form of losses and gains of chromosome 17 were diagnosed in 9 and 7 patients, respectively. There was no correlation with tumor prognosis, clinical stage or histologic grade. According to most reports, almost all cervical carcinomas contain integrated human papilloma virus (HPV) and express E6 oncoproteins. Increasing evidence suggests that E6 protein interaction leads to p53 mutation in HPV infected cervical epithelium. Since most cervical tumors are infected with HPV, and the tumors originate through p53 gene mutation caused by the said interaction, which leads subsequently to the overexpression of p53 oncoprotein, lack of the latter in the remaining 11 cervical tumors may either be the result of technical shortcomings, or the tumor may arise in such circumstances through a p53-independent pathway. On the other hand, 2 of 3 stage IIa cancers and 1 Ib G3 carcinoma were found to be p53 positive, thus supporting the notion that p53 inactivation is a relatively late event in the progression of cervical cancer. PMID- 11408734 TI - Genetic alterations in microsatellite marker sites among tumor suppressor genes in endometriosis. AB - Four endometriotic lesions were examined for the presence of genetic alterations in microsatellite marker sites among eight tumor suppressor genes. For this, a microdissection method was used on paraffin sections. Only one instance of loss of heterozygosity was detected at the PTCH locus. Heterozygosity was retained (indicating the absence of both loss of heterozygosity and microsatellite instability) at the other seven tumor suppressor gene loci in all the cases. Among the tumor suppressor genes examined, genetic defects in these microsatellite regions are certainly not ubiquitous in endometriosis and may be uncommon. PMID- 11408735 TI - Long-term results of Burch colposuspension. AB - The study included 220 women who had undergone Burch colposuspension. Group I (65 women) was studied prospectively and the mean follow-up was 1.5 years. Group II (155 women) was studied retrospectively and the mean follow-up was 4.5 years. The cure rate was 87.7% in group I and 77.4% in group II. The cure rate was significantly higher following the primary procedure than the secondary procedure. At follow-up, late complications in 220 women were: cystocele in 18; rectocele in 32; enterocele in 35; dyspareunia in 6, and groin or suprapubic pain in 15. In group I, of the 11 women with detrusor instability preoperatively, 10 were cured and in 1 detrusor instability persisted postoperatively. Two women had de novo detrusor instability. In conclusion, the cure rate of Burch colposuspension is satisfactory, although it declines a little with time. Women who had previous anti-incontinence surgery have a greater probability of recurrence. The procedure elevates the bladder neck into the abdominal cavity and stabilizes it. Surgical failure is related to inadequate elevation and stabilization of the bladder neck. PMID- 11408736 TI - Potential role of beta 1 integrin and collagen biosynthesis in estrogen-dependent reduction of apoptosis in tamoxifen-treated breast cancer cells. AB - It was found that 10 microM tamoxifen induced apoptosis and a significant (approximately 50%) depletion of beta 1 integrin levels in human breast cancer cells. Estradiol-treated MCF-7 cells exhibited exceptional viability and adherence, high levels of beta 1 integrin and increased (by 100%) collagen biosynthesis. Pretreatment of MCF-7 cells with 1 nM estradiol prevented tamoxifen induced cell death, loss of cell adherence and decrease in beta 1 integrin level. Tamoxifen and estradiol had an opposite effect on the beta 1 integrin level and adherence in breast cancer cells, suggesting that the decrease in the beta 1 integrin level may be an early event during tamoxifen-induced apoptosis in breast cancer cells. PMID- 11408737 TI - Phenotypic and functional analysis of tumor-infiltrating lymphocytes compared with tumor-associated lymphocytes from ascitic fluid and peripheral blood lymphocytes in patients with advanced ovarian cancer. AB - To investigate and compare the phenotype and function of lymphocytes collected from patients harboring advanced ovarian cancer, leukocytes from peripheral blood (n = 18), ascitic fluid (n = 13) and tumor tissues (n = 13) were evaluated for the relative proportions of lymphocyte subsets, including CD3+, CD4+, CD8+, CD19+, CD56 and the early (CD25) and late (HLA-DR) activation markers on CD3+ T cells. The ability to synthesize type 1 cytokines (IFN-gamma and IL-2) and a type 2 cytokine (IL-4) was assessed by flow cytometry. In all patients, T cells (CD3+) were the major leukocyte population detected in each tissue, with CD4+ T cells being dominant in peripheral blood lymphocytes (PBL) and tumor-associated lymphocytes (TAL) but not in tumor-infiltrating lymphocytes (TIL) (CD4:CD8 ratios: 3.0 vs. 2.0 vs. 1.0, respectively). CD19+ lymphocytes (B cells) and CD56+ lymphocytes (NK cells) were significantly higher in PBL compared to TAL and TIL (p < 0.05). TAL and TIL had a higher proportion of T cells expressing the late activation marker HLA-DR compared to PBL. In contrast, no significant differences were detected in PBL, TAL and TIL in the expression of the early activation marker CD25. Type 1 cytokines were the dominant type produced by in vitro stimulated T cells for each population, with a greater proportion of IFN-gamma+ T cells in TAL and TIL compared to PBL (p < 0.01), and a higher proportion of IL-2+ T cells in PBL compared with TAL and TIL (p < 0.05). Low percentages of IL-4+ T cells (i.e. Th2) were detected in each tissue. Taken together, these data demonstrate the recruitment and accumulation of high concentrations of antigen experienced T lymphocytes in TAL and TIL compared to PBL. However, low surface expression of IL-2 receptor (i.e. CD25), as well as depressed intracellular IL-2 production in chronically stimulated TAL and TIL suggests that the impaired antitumor function commonly detected in these lymphocyte populations may be secondary to an acquired dysregulation of the IL-2 pathway. PMID- 11408738 TI - Prognostic factors for survival in invasive squamous cell vulvar carcinoma: a univariate analysis. AB - The charts of 50 women with invasive squamous cell vulvar carcinoma were reviewed retrospectively, and pathologic, medical and life style factors were analyzed for their possible influence on survival using the Cox regression univariate model. The median age of the cohort was 73.5 years. The patient distribution according to stage was as follows: stage I: 17; stage II: 16; stage III: 12, and stage IVa: 5 patients. The median follow-up was 61 months. The univariate analysis revealed that the overall survival was decreased by age > or = 73.5 years (p = 0.0185), advanced stage (p = 0.0026), grade III differentiation (p < 0.0001), ulcerative type of the tumor (p = 0.0055), tumor diameter >40 mm (p = 0.0053), obesity (p = 0.011), smoking (p = 0.0177), diabetes (p = 0.0122) and hypertension (p = 0.044), but not with clitoral involvement. PMID- 11408739 TI - Pregnancy in hyperprolactinemic infertile women treated with vaginal bromocriptine: report of two cases and review of the literature. AB - Vaginal bromocriptine has proven safe and effective in treating hyperprolactinemic women. However, there has been no long-term clinical assessment regarding the influence of daily vaginal bromocriptine administration on the ability to conceive. This article presents two cases of successful pregnancy resulting from this alternative treatment. An infertile woman with an empty sella and hyperprolactinemia was treated with vaginal bromocriptine because of intolerance to oral administration. Prolactin levels were quickly normalized and no side effects occurred. Repeated postcoital tests during treatment proved normal. Twelve months later, the patient conceived. The therapy was discontinued during pregnancy, without complications. Although bromocriptine treatment was not resumed after delivery, postpartum prolactin levels were lower than before treatment and magnetic resonance imaging revealed an unchanged empty sella. Another patient with infertility and pituitary microadenoma with intolerance to oral dopaminergic agonists received the same treatment. Prolactin quickly fell to within the normal range. Vaginal bromocriptine was well tolerated and postcoital test results were not impaired. Tumor regression occurred and 10 months later the patient conceived. Despite bromocriptine withdrawal, no significant complications occurred during pregnancy. It can therefore be concluded that a couple's fertility does not appear to be significantly affected by the persistent local presence of bromocriptine. PMID- 11408740 TI - Vaginal adenosis in a non-diethylstilbestrol-exposed 6-year-old patient. AB - Vaginal adenosis is rare, and it is defined as the presence of metaplastic cervical or endometrial epithelium within the vaginal wall. It is associated with in utero exposition to diethylstilbestrol and a high risk of vaginal carcinomas. A case of vaginal adenosis arising in a non-diethylstilbestrol-exposed 6-year-old patient is presented. Few cases have been described in children and adolescents, and since the withdrawal of diethylstilbestrol from the market, this condition is rarely described in the medical literature. However, it should be considered as a possible diagnosis in girls with persistent vaginal discharge. PMID- 11408741 TI - Disseminated intravascular coagulation associated with intratumoral hemorrhage of ovarian cancer. AB - BACKGROUND: Ovarian carcinoma usually presents in an indolent manner. A patient with ovarian cancer rarely shows acute clinical features which require immediate medical intervention. CASE: We present a 36-year-old Japanese woman with an ovarian tumor, who suffered general fatigue and increased abdominal girth on admission. Her hemoglobin, platelet count and fibrinogen were decreased, and fibrin/fibrinogen degradation products were elevated, which determined the diagnosis of disseminated intravascular coagulation (DIC). Since computed tomography scan and magnetic resonance imaging revealed an intratumoral hemorrhage, an emergency laparotomy was performed. The patient was found to have ovarian cancer with a massive intratumoral hemorrhage, and DIC improved immediately after the operation. CONCLUSION: The clinician should be alert to the possibility that intratumoral bleeding of ovarian cancer might be associated with DIC. PMID- 11408742 TI - Genital tuberculosis can present as disseminated ovarian carcinoma with ascites and raised Ca-125: a case report. AB - In women with an adnexal mass, ascites and elevated Ca-125 levels, ovarian carcinoma must be ruled out. However, several other conditions, including genital tuberculosis, may present similarly. A 41-year-old woman with weight loss, ascites and elevated levels of Ca-125 was evaluated for ovarian cancer. Computerized tomography revealed an adnexal mass, ascites and lymph nodes on the peritoneal surface. Paracentesis of the ascitic fluid revealed a lymphocytic exudate but failed to show any malignant cells. At laparotomy, frozen sections of tissue biopsies were negative for malignancy; however, a total hysterectomy plus adnexectomy was performed. Postoperatively histologic examination revealed typical features of genital tuberculosis. Antituberculosis treatment was effectively given to the patient. Serum levels of Ca-125 were undetectable 12 weeks after treatment. In conclusion, genital tuberculosis can be misdiagnosed and confused with ovarian cancer. Intraperitoneal tuberculosis should be considered in the differential diagnosis in cases in which ovarian cancer is suspected, even when malignancy-associated risk factors are present. PMID- 11408743 TI - Alpha-2-macroglobulin inhibits the anticoagulant action of activated protein C in cord and adult plasma. AB - Healthy newborns have a very low risk of thrombosis. It has been suggested that this is partly due to the anticoagulant effect of alpha-2-macroglobulin (a2-M). This broad-spectrum protease binding glycoprotein is physiologically elevated in newborns over adult values and has been shown to complex generated alpha thrombin. In our present study, we point out that a2-M also acts as a procoagulant by inhibiting activated protein C (APC). In all experiments performed in cord and adult plasma the anticoagulant action of APC was diminished in a dose-dependent manner when a2-M levels were successively elevated, reflected in increased thrombin potential (TP), and enhanced at low a2-M levels, reflected in decreased TP. PMID- 11408744 TI - Fibrinogen kaiserslautern III: a new case of congenital dysfibrinogenemia with aalpha 16 arg-->cys substitution. AB - An abnormal fibrinogen was identified in a man with suspicious prolonged prothrombin time and a mild bleeding tendency. Coagulation studies showed marked prolonged thrombin and reptilase clotting times and a discrepancy between functional fibrinogen test and fibrinogen antigen. The rate of fibrinopeptide B release by thrombin was slightly delayed while the release of fibrinopeptide A was only half the normal amount. DNA sequencing revealed a heterozygous C to T point mutation in position 1202 of exon 2 of the Aalpha chain, resulting in the substitution of Arg-->Cys at position 16, the thrombin cleavage site. This mutation was found also in his 2 children. Both had a mild bleeding tendency too. PMID- 11408745 TI - Comparative study of a portable prothrombin time monitor employing three different systems in oral anticoagulant units. AB - The aim of this study was to evaluate the accuracy of the portable coagulometer CoaguChek (Roche Diagnostics) as a prothrombin time (PT) monitor, and to correlate capillary blood results with those of three different routine methods used for monitoring oral anticoagulant therapy (OAT): capillary, plasma and whole blood samples. Three hospitals participated in the study with a total of 235 patients on OAT. The international normalized ratio (INR) results obtained with CoaguChek were compared with those obtained using each of the routine methods. The study presents a good correlation between the PT monitor and the three methods studied: r = 0.9745 (hospital A), r = 0.9283 (hospital B), r = 0.9136 (hospital C). A simplified concordance test of the methods results in a nine field comparison table showing concordances of 87.2, 85.7 and 68.4%, respectively. The absolute difference (mean +/- SD) between laboratory A and CoaguChek INRs was 0.0571 +/- 0.2042, with values of 0.04286 +/- 0.3906 for laboratory B and 0.6986 +/- 0.6170 for laboratory C. These results confirm that CoaguChek could be used as a new method for oral anticoagulant monitoring, and is in best agreement with the capillary blood PT system. PMID- 11408746 TI - Measurement of a newly developed thrombomodulin addition activated partial thromboplastin time assay in patients with deep venous thrombosis. AB - We developed a simple assay using rabbit thrombomodulin (TM) based on an activated partial thromboplastin time method, which detected the response to TM in plasma coagulation. We call it thrombomodulin addition clotting time (TACT). The anticoagulant response to TM was calculated by dividing the clotting time with TM by the clotting time with buffer solution. Results were expressed as TACT ratio, which indicates the degree of inhibition of plasma clotting by TM. Using this assay, we measured the TACT ratio in 80 patients with deep-vein thrombosis (DVT) and in 126 controls matched to the patients according to age and sex. A significant difference in the TACT ratio was observed between patients with DVT (mean 1.874) and controls (mean 1.956) (p < 0.001). Twenty- three patients (29%) had TACT ratios below the 10th percentile (1.757) of distribution of control subjects (odds ratio: 3.5; 95% confidence interval (CI): 1.7-7.2). After excluding subjects with a deficiency of protein C, protein S and antithrombin III, we found an odds ratio for DVT of 3.4 (95% CI: 1.6-7.2). These data suggest that natural anticoagulant deficiencies do not influence the TACT ratio, and our case-control study may show that the plasma of patients with DVT has a low response to TM. PMID- 11408747 TI - Changes in haemostasis after laparoscopic surgery in gynaecology: contribution of the thrombin generation test. AB - Surgery induces immediate hypercoagulability by direct alteration of the vascular bed, release of procoagulant substances from the extravascular spaces and blood flow decrease, and delayed hypercoagulation in response to tissue damage which triggers inflammatory responses. Thus, the postoperative period represents a high risk time for thrombosis. Recognition of high-risk individuals would make it possible to improve thromboembolism prevention. We studied in women undergoing laparoscopic surgery a series of markers known to be related to the thrombotic risk and confronted their results with those of a global test, the thrombin generation test (TGT) described by Hemker's group. Our results show that two groups of patients can be distinguished according to usual risk markers (PAI-1, TAT, body mass index): the higher risk group demonstrates higher initial TGT values, but also a postoperative decrease of the TGT values whose mechanisms remain to be defined. PMID- 11408748 TI - Effect of patient weight on the anticoagulant response to adjusted therapeutic dosage of low-molecular- weight heparin for the treatment of venous thromboembolism. AB - Data evaluating the safety of using weight-based dosing of low-molecular-weight heparin (LMWH) in obese patients are limited. Some manufacturers have recommended a maximum daily dose of LMWH not to be exceeded. The purpose of this study was to determine if body weight influenced the anticoagulant response to a weight-based dose of LMWH for the treatment of venous thromboembolism. Patients with serum creatinine levels < 150 micromol/l receiving the LMWH, dalteparin 200 anti-Xa IU/kg based on actual body weight subcutaneously once daily for the treatment of deep vein thrombosis or pulmonary embolism, were eligible for the study. Patients received a minimum of 5 days LMWH treatment. Patients had peak anti-Xa levels (IL Test Chromogenic assay) measured 3-4 h following their day 3 injection and trough anti-Xa levels measured immediately prior to injections on day 3 and 5. No dose adjustments were made on the basis of the anti-Xa levels. Patients were a priori stratified into three weight classes: (A) within 20% of ideal body weight (IBW) (n = 13); (B) 20-40% of IBW (n = 14), and (C) greater than 40% of IBW (n = 10). The largest patient weighed 190 kg and had a body mass index of 58. Mean daily LMWH doses were 14,030, 17,646 and 23, 565 IU for groups A, B and C, respectively. Mean (SD) trough anti-Xa levels on day 3 were 0.12 (0.05) anti-Xa IU/ml for group A, 0.11 (0.03) anti-Xa IU/ml for group B and 0.11 (0.03) anti-Xa IU/ml for group C (p > 0.2). Similar trough anti-Xa levels were observed on day 5. Mean (SD) peak anti-Xa levels on day 3 were 1.01 (0.20) anti-Xa IU/ml, 0.97 (0.21) anti-Xa IU/ml and 1.12 (0.22) anti-Xa IU/ml for groups A, B and C, respectively (p > 0.2). No thromboembolic or bleeding complications occurred during LMWH therapy in any patients. These findings suggest that body mass does not appear to have an important effect on the response to LMWH up to a weight of 190 kg in patients with normal or near normal renal function. PMID- 11408749 TI - Increased indexes of thrombin activation in advanced stages of hypertension. AB - BACKGROUND: The hemostatic system plays an important role in thrombotic lesions, which can complicate the clinical course of hypertensive patients. The aim of this study was to verify a possible activation of the blood clotting processes, the evaluation of two markers of thrombin activation in 62 hypertensive patients, with and without vascular complications, compared with a control group. METHODS AND RESULTS: In 22 patients with newly diagnosed uncomplicated essential hypertension, in 40 hypertensive patients with clinically evident vascular complications (20 patients with controlled blood pressure and 20 with uncontrolled and high blood pressure) and in 20 normotensive sex- and age-matched subjects, two indexes of thrombin generation and action, namely prothrombin fragment 1 + 2 (F1 + 2) and fibrinopeptide A (FPA) were evaluated. The observed values show an increase of the F1 + 2 levels in patients with overt vascular complications; those with higher blood pressure, moreover, showed FPA levels higher than those with controlled blood pressure. CONCLUSIONS: These results seem to indicate that plasma F1 + 2 levels are significantly elevated, as a marker of a thrombosis-prone status, in patients with organic damage. Successively, with progress of hypertension and increasing blood pressure, the evidence of elevated FPA levels seems to indicate a clear prethrombotic situation which could turn into a thrombotic state. PMID- 11408750 TI - Inverse relationship between platelet density and reactivity alterations at coronary angiography. AB - This work investigates relationships between platelet density and reactivity. 21 individuals subject to coronary angiography were studied. Peak platelet density was analyzed using a newly developed electronic device. The apparatus measures light transmission through test tubes containing density-separated platelets, thus allowing an estimation of the platelet distribution in the gradient. A flow cytometry technique was used for determining platelet reactivity after stimulating with ADP. Platelet counts, mean platelet volumes, peak platelet density and platelet reactivity were determined immediately before (day 1) and 24 h after cardiac catheterization (day 2). For all parameters changes during the day of angiography were compared with platelet density alterations. The subjects were divided into two groups according to density changes at angiography. Group 1 individuals showed density alterations (i.e. day 2 - day 1 value) > or = -8 x 10( 5) kg/l. In contrast, group 2 subjects either displayed density changes < -8 x 10(-5) kg/l or grossly disturbed platelet density patterns on day 2. Before angiography both groups had similar platelet counts and volumes. Then platelet reactivity when stimulating with ADP did not differ significantly between the two groups. After angiography, the number of fibrinogen-positive cells when stimulating with ADP rose by 6 +/- 8% for group 2 patients. The corresponding figure for group 1 was -1 +/- 6%. The difference was significant (p = 0.01). No such relationships were found when comparing density alterations and changes of platelet counts and volumes. We conclude that in this study platelet density alterations at coronary angiography are inversely related to variations of platelet reactivity. PMID- 11408751 TI - Preanalytical recommendations of the 'Groupe d'Etude sur l'Hemostase et la Thrombose' (GEHT) for venous blood testing in hemostasis laboratories. PMID- 11408752 TI - Low-molecular-weight heparins, acenocoumarol and bone density. PMID- 11408753 TI - Possible roles of insulin-like growth factor in regulation of physiological and pathophysiological liver growth. AB - BACKGROUND/AIMS: Almost all circulating insulin-like growth factor-1 (IGF-1) is produced and secreted from the liver. However, the possible role of IGF-1 in local regulation of liver functions including liver growth is unclear. In the present study, we investigated the role of IGF-1 on liver growth in vivo and in hepatic stellate cell function in vitro. RESULTS: Liver-specific knock-out of the IGF-1 gene by use of the cre-loxP system caused enhanced liver growth, possibly reflecting increased growth hormone (GH) secretion due to decreased negative feedback by IGF-1. Studies on cultured rat hepatic stellate cells (HSC) showed that IGF-1 and hepatocyte-conditioned medium (PCcM) time- and dose-dependently increased hepatocyte growth factor (HGF) mRNA and HGF immunoreactivity. IGF-1 and PCcM also enhanced DNA synthesis in the HSC cultures. The PCcM did not contain bioactive IGF-1 and was also able to stimulate proliferation when prepared under serum- and hormone-free conditions. CONCLUSION: In vivo results show that IGF-1 is not essential for normal growth of the intact liver. The in vitro results indicate that both IGF-1 and IGF- 1-independent factor(s) from hepatocytes can stimulate HGF production by HSC. It remains to be investigated whether these effects are of importance for liver regeneration or pathological conditions. PMID- 11408754 TI - Insulin resistance and insulin-like growth factors in children with intrauterine growth retardation. Is catch-up growth a risk factor? AB - AIMS: To investigate (a) the prevalence of insulin resistance in children with intrauterine growth retardation (IUGR); (b) whether catch-up growth is associated with a higher risk of insulin resistance; (c) the insulin-like growth factor (IGF) system status. METHODS: 49 children with IUGR aged 9.1 +/- 3.3 years underwent anthropometric measurements, and assessment of insulin resistance and IGF system parameters. A fasting glucose/insulin ratio (G/I) <6 was chosen as suggestive of insulin resistance. RESULTS: 11/49 (22%) children had a G/I <6. Postnatal growth closely correlated with birth size and actual body mass index (BMI). None of the insulin resistance parameters was related to linear growth and BMI. Liver function markers were significantly related to insulin sensitivity status. The IGF system status was normal and did not correlate with insulin resistance indicators. CONCLUSIONS: (a) Children with IUGR have a high prevalence of reduced insulin sensitivity; (b) postnatal catch-up growth is related to intrauterine growth and actual nutritional status; (c) insulin sensitivity status is not related to postnatal growth but to liver function; (d) IGF system is normal and not related to the insulin resistance parameters during childhood. PMID- 11408755 TI - Hormonal control of growth hormone secretion. AB - Growth hormone secretion by the somatotroph cells depends upon the interaction between hypothalamic regulatory peptides, target gland hormones and a variety of growth factors acting in a paracrine or autocrine fashion. This review will be focused on recent data regarding the mechanism by which growth hormone-releasing hormone (GHRH) influences somatotroph cell function and the physiological role played by Ghrelin and leptin in the regulation of growth hormone (GH) secretion. It is well established that binding of GHRH to its receptor leads to activation of protein kinase A (PKA). More recently, it was found that GHRH can also activate mitogen-activated protein (MAP) kinase both in pituitary cells and in a cell line overexpressing the GHRH receptor. Whether somatotroph adenomas, either with or without a GS-alpha mutation, have alterations in some of the components of the activation of the MAP kinase pathway remains to be known. The recent isolation of Ghrelin, the endogenous ligand of the growth hormone secretagogue receptor, can be considered a landmark in the GH field, which opens up the possibility of gaining greater insight into our understanding of the mechanisms involved in the regulation of GH secretion and somatic growth. Indeed, preliminary evidences indicate that this peptide exerts a marked stimulatory effect on plasma GH levels in both rats and humans. Finally, it is well known that GH secretion is markedly influenced by nutritional status. Leptin has emerged as an important adipose tissue-generated signal that is involved in the regulation of GH secretion, thus providing an integrated regulatory system of growth and metabolism. Although the effects of leptin on GH secretion in humans remain to be clarified, indirect evidences indicate that it may play an inhibitory role. PMID- 11408756 TI - Insulin-like growth factor -I deficiency. AB - The insulin-like growth factor (IGF) system composed of two ligands, their receptors and regulatory proteins (acid-labile subunit and IGF-binding proteins) plays a central role in the regulation of growth and development in mammals. In addition to its key role in the stimulation of cellular proliferation and growth, IGF-I has important effects on carbohydrate, protein and bone metabolism. The molecular biology and physiology of the IGF system are complex, resulting in many potential mechanisms of IGF deficiency. Briefly, IGF-I deficiency may result from a primary defect in the IGF-I gene, its promoters, or may be secondary to a defect outside the gene itself. It may also result as a consequence of growth hormone (GH) deficiency, GH receptor/post-receptor abnormalities or abnormalities of the IGF-I receptor. The purpose of this presentation is to review the different types of IGF-I deficiency using the well-characterized clinical conditions with its associated biochemical and molecular defects. The clinical consequences in terms of phenotype-genotype, linear growth and body composition in patients with primary and secondary IGF deficiency will be presented, together with results from recombinant human (rh)IGF-I replacement therapy. Finally, as primary IGF-I deficiency is associated with insulin resistance, some of the metabolic actions of IGF-I will be briefly discussed. PMID- 11408757 TI - SHOX in short stature syndromes. AB - Linear growth is a multifactorial trait that is influenced and regulated by a combination of environmental and internal factors. Among the intrinsic determinants of final body height, genetic factors have become more and more prominent, and the list of genes involved in growth-related processes has been extended accordingly. One of the most exciting additions to this list is represented by the discovery of the pseudoautosomal gene SHOX. Originally described as a gene responsible for idiopathic short stature, it has become clear that SHOX mutations can also cause mesomelic short stature and Madelung deformity in Leri-Weill syndrome. In addition, recent studies implicate SHOX haploinsufficiency in a variety of somatic Turner syndrome stigmata. PMID- 11408758 TI - Cushing's disease in childhood: presentation, investigation, treatment and long term outcome. AB - Seventeen patients with Cushing's disease (CD) were treated from 1978 to 2000. There were 11 males and 6 females aged 6.8-18.8 years (mean age 13.0 +/- 5.9 years). Presenting features were: weight gain (100%); growth failure (71%); hirsutism (53%); striae (53%); hypertension (47%). Mean age of patients with striae was 15.2 +/- 2.3 years, without striae 10.3 +/- 3.3 years. Median height SDS was -1.81 (range -0.28 to -4.17), 53% having height SDS < -1.8. The height velocity in 6 subjects was subnormal (0.9-3.8 cm/year). Median BMI SDS was 2.29 (range 1.72-5.06). Cushing's disease was confirmed by detectable serum ACTH, median 28 ng/l (range 12-99, NR <10-50) (n = 15); loss of cortisol circadian rhythm values at midnight ranging from 216 to 1,080 nmol/l (NR <50) (n = 15); lack of cortisol suppression (NV < 50 nmol/l) during low-dose dexamethasone suppression test (LDDST) (0.5 mg 6-hourly x 8) (n = 14); and >50% suppression of cortisol compared with the basal value during high-dose dexamethasone suppression test (HDDST) (2 mg 6-hourly x 8) (n = 14). A CRH test (1 microg/kg i.v.) showed an increase of cortisol from 12 to 217% (median 73.5%) (n = 16). Pituitary imaging (CT/MRI) showed an image consistent with microadenoma in 6/17 patients, but there was concordance between pituitary imaging and surgical findings in 1/11 patients (9%). Inferior petrosal sinus sampling (IPSS) for ACTH after CRH was performed in 11 subjects (age 10.7-18.8 years). Central to peripheral ACTH ratios were >2 (2.5-157.2) in 10/11 patients. The inter-petrosal sinus ACTH gradient was >1.4 in 10 patients (2.1-20.8), indicating lateralization of ACTH secretion. In 10 patients (91%), the side of the tumour on IPSS was predictive of findings at surgery. Therapy consisted of transsphenoidal microadenomectomy (TSS) in 16 patients and bilateral adrenalectomy (1978) in 1. Following TSS alone, 7 patients were cured (cortisol <50 nmol/l) and 2 were in remission (cortisol <300 nmol/l), i.e. 56%. Seven had persisting hypercortisolaemia and underwent pituitary irradiation (4,500 cGy). Therapeutic outcome for a median of 8 years (0.5-24 years) resulted in cure of CD in 14/17 patients (82%) and remission in 1. Linear growth after TSS +/- pituitary irradiation in 10 subjects showed no short-term catch-up growth, with peak growth hormone (GH) 0.5-20.9 mU/l to insulin tolerance test (ITT)/glucagon. Eight patients were treated with human growth hormone (hGH) (14 U/m(2)/week) combined in 3 with GnRH analogue. The mean final (n = 6) or latest (n = 4) height SDS was -1.36. The difference between final/latest height SDS and target height SDS was 0.93 +/- 1.13, i.e. less (p = 0.005) than the difference between height SDS and target height SDS at presentation, i.e. 1.72 +/ 1.26, indicating long-term catch-up growth. PMID- 11408759 TI - Investigating familial endocrine neoplasia syndromes in children. AB - Familial endocrine neoplasia syndromes multiple endocrine neoplasia (MEN) type 1, MEN type 2 and von Hippel Lindau (VHL) can now be diagnosed genetically in childhood. Paediatric endocrinologists must therefore be prepared to investigate and manage these children. This paper provides an overview of the major features of these syndromes and suggests protocols for regular screening of children known to be at risk of developing these disorders. PMID- 11408760 TI - Metabolic syndromes in human immunodeficiency virus infection. AB - Infection with human immunodeficiency virus (HIV) is associated with marked disturbance of metabolism affecting the metabolism of carbohydrates, fats and proteins. In the first decade of clinical experience of HIV, the primary clinical manifestation of such disturbed metabolism was wasting. Such wasting was often severe and contributed significantly to the morbidity and mortality of AIDS. Mechanistic studies demonstrated that in addition to the effects of altered intermediary metabolism, reduced food intake played a major role in the causation of AIDS-related wasting. More recently, potent anti-retroviral drugs have dramatically changed the clinical consequences of HIV infection. Wasting has become far less frequent among infected patients and occurs in only a small percentage of subjects on effective anti-retroviral therapy. However, a new constellation of metabolic syndromes has become apparent characterized by altered body fat distribution ('lipodystrophy'), lactic acidosis and evidence of mitochondrial dysfunction. The mechanistic basis for such syndromes is currently unclear, but is the subject of ongoing research. PMID- 11408761 TI - Long-term effects of childhood obesity on morbidity and mortality. AB - Obesity tracks from childhood into adulthood, and the persistence of obesity rises with age among obese children. Early onset obesity was suggested as a risk factor for morbidity and mortality later in life. In both sexes, rates of diabetes, coronary heart disease, atherosclerosis, hip fracture and gout were increased in those who were overweight as adolescents. Especially in females, obesity at late adolescence was associated with several and relevant psychosocial consequences in adulthood. Finally, a higher mortality risk for all causes of death, especially atherosclerotic cerebrovascular disease and colorectal cancer, was demonstrated in males but not in females who were overweight during high school years. Although the persistence of excess adiposity from childhood to adulthood is a morbidity risk factor, it is not known if total body fat or body fat distribution is the main factor responsible. In particular, a specific role for the intra-abdominal adipose tissue (IAAT) in childhood, independently from that of total body fat, on morbidity risk in adulthood was not demonstrated yet. The association between childhood obesity and adult morbidity and mortality strongly suggests that a more effective prevention and treatment of childhood obesity should be pursued. PMID- 11408763 TI - Gene expression accompanied by differentiation of cord blood-derived CD34+ cells to eosinophils. AB - To clarify the relation between the expression of genes such as eosinophil specific granular proteins and cytokine receptors and the pathogenesis of allergic disease, cord blood-derived CD34+ cells were cultured and differentiated into eosinophils. Gene expression in the cells during the differentiation was determined by real-time reverse transcription PCR (ABI PRISM 7700). CD34+ mononuclear cells cultured with stem cell factor, granulocyte-macrophage colony stimulating factor (GM-CSF), IL-3, and IL-5 in Iscove's MEM, and proliferated until the 2nd week, when the cell number reached a plateau. Under these conditions, more than 90% of the cells differentiate into mature eosinophils in 3 weeks. The expression of major basic protein and eosinophil-derived neurotoxin in the treated cells increased until week 2 and decreased between week 2 and 3. However, the expression of membrane receptor genes, such as IL-5 receptor (alpha chain), IL-3 receptor (alpha chain), GM-CSF-alpha receptor, GM-CSF-beta receptor, CC chemokine receptor 3, interferon-gamma receptor, platelet-activating factor receptor and leukotriene D4 receptor, increased until the 3rd week of eosinophil maturation. Our study suggests that the in vitro eosinophil differentiation and maturation model is useful for clarifying the relation between eosinophil specific gene expression during allergic diseases and the progression of the disease. PMID- 11408764 TI - Eosinophil transmigration across VCAM-1-expressing endothelial cells is upregulated by antigen-stimulated mononuclear cells. AB - BACKGROUND: Adhesion to and transmigration across endothelial cells expressing vascular cell adhesion molecule 1 (VCAM-1) may be key steps in the development of selective eosinophil accumulation at the allergic inflammation sites. There is evidence that cytokines/chemokines produced by CD4+ T cells play a prominent role in these processes. OBJECTIVE: The objective of this study was to evaluate whether eosinophil migration across human pulmonary microvascular endothelial cells (HPMEC) expressing VCAM-1 is modulated by the supernatants of antigen stimulated mononuclear cells obtained from atopic asthmatics. METHODS: Peripheral blood mononuclear cells (PBMC) were isolated from Dermatophagoides farinae (Df) sensitive asthmatic subjects and cultured for 96 h at 37 degrees C in the presence or absence of 1 microg/ml Df antigen. Eosinophils were isolated from blood of healthy subjects and placed on the HPMEC monolayers cultured on a transwell filter (3-microm pore size) stimulated with IL-4 plus TNF-alpha (both at 100 pM, 24 h) The supernatants of PBMC were then applied to the lower compartment and the transmigration of eosinophils was examined. RESULTS: The supernatants of PBMC stimulated with Df significantly enhanced the eosinophil transmigration across VCAM-1-expressing HPMEC (% migration: 7.6 +/- 0.6 by the supernatants of PBMC cultured without Df vs. 12.3 +/- 1.2 by the PBMC cultured with Df, p < 0.01, n = 8). The enhanced migration, but not spontaneous migration, was blocked by the anti-alpha4 integrin antibody. Moreover, the enhanced transmigration was blocked by anti-CCR3 antibody. CONCLUSION: The antigen stimulated PBMC from atopic asthmatics produce an activity to induce eosinophil migration across VCAM-1-expressing endothelial cells. This activity appears to involve CCR3 as an essential molecule. PMID- 11408765 TI - Dexamethasone does not modulate eosinophil adhesion to endothelial cells. AB - BACKGROUND: Glucocorticoids are representative agents in the anti-inflammatory treatment of allergic airway diseases. Although they reduce the number of tissue eosinophils, its exact mechanism has not been completely established. OBJECTIVE: The present study was undertaken to evaluate whether dexamethasone modulates the adhesive property of eosinophils. The effect on the expression of endothelial adhesion molecules was also evaluated. METHODS: Blood eosinophils from healthy subjects were incubated in the presence or absence of 0.1-1 microM of dexamethasone for 6 h at 37 degrees C. Eosinophil chemotaxis and adhesion to paraformaldehyde-fixed human pulmonary vascular endothelial cells (HPMEC) were then examined. The effects of dexamethasone on the expression of VCAM-1 and ICAM 1 on HPMEC were examined by cell ELISA. RESULTS: 1 microM dexamethasone significantly inhibited the chemotactic response of eosinophils to 1 microM formyl-methionyl-leucyl-phenylalanine (FMLP). On the other hand, dexamethasone did not modify either the spontaneous or FMLP-stimulated adhesion of eosinophils to resting HPMEC. Similarly, dexamethasone did not alter either the spontaneous adhesion of eosinophils to or the expression of VCAM-1 or ICAM-1 on HPMEC stimulated with IL-4 + TNF-alpha. CONCLUSION: Dexamethasone does not modify the adhesive property of eosinophils or the expression of adhesion molecules on endothelial cells. Thus, it is unlikely that glucocorticoids directly modulate the adhesive interaction between eosinophils and endothelial cells. PMID- 11408766 TI - Humoral immunity is dispensable for the local recruitment of antigen-specific Th2 cells. PMID- 11408767 TI - Chemotaxis of human CD4+ eosinophils. AB - We reviewed the biological functions of CD4+ eosinophils, which are observed in peripheral blood, sputum and bronchoalveolar lavage fluid of various diseases. We have shown that CD4 molecules on human eosinophils are induced by tumor necrosis factor-alpha (TNF-alpha) stimulation. Interleukin-16 (IL-16) has been reported to bind a natural soluble ligand for the CD4 molecule. We reported that TNF-alpha stimulated eosinophils migrate in a time- and dose-dependent manner against IL 16. Theophylline and dexamethasone significantly inhibited CD4 expression. Theophylline inhibited CD4+ eosinophil chemotaxis, but dexamethasone did not. Theophylline may prevent airway inflammation by downregulating the expression of CD4 molecule and CD4+ eosinophil migration. However, dexamethasone may inhibit airway inflammation through the downregulation of CD4 expression. PMID- 11408768 TI - Effect of emedastine difumarate on CC chemokine-elicited eosinophil migration. AB - Emedastine difumarate (emedastine) is an antiallergic drug found among the derivatives which has a series of benzimidazole frames. It has been reported that emedastine can significantly inhibit the migration of eosinophils elicited by classical chemoattractants, including LTB4 or PAF. However, the effect of emedastine on the selective migration of eosinophils that have been stimulated with CC chemokines has not been examined. Emedastine at concentrations of 10 nM or higher strongly inhibited the eosinophil migration elicited by CC chemokines, including eotaxin, RANTES and MCP-3. Preincubation of the eosinophils with emedastine did not alter the expression of the CCR3 receptor, although a decrease in the concentration of intracellular calcium ions was observed after stimulation with 100 ng/ml of eotaxin. Herbimycin A, genistein, staurosporin and emedastine were all able to inhibit the eotaxin-elicited migration. Tyrosine kinase activity in the cytosol supernatant of eosinophils obtained after stimulation with eotaxin significantly decreased when the eosinophils were preincubated with emedastine. In addition, protein kinase A and protein kinase C activities in eotaxin stimulated EoL-1 cell supernatants decreased significantly with emedastine pretreatment. These findings suggest that emedastine inhibits CC chemokine elicited eosinophil migration by decreasing the activities of tyrosine kinase or protein kinases but does not alter CCR3 expression. PMID- 11408769 TI - Regulation of chemokine receptor expression in eosinophils. AB - Signals via chemokine receptors play an important role in the accumulation of eosinophils at allergic inflammatory sites. Eosinophils constitutively express CC chemokine receptor 3 (CCR3) and, to a lesser extent, CCR1. CCR3 is mainly responsible for migration of resting eosinophils, and its specific ligand, eotaxin, represents the most potent chemoattractant for eosinophils. Some reports also suggest the expression of CXC chemokine receptor 1 (CXCR1) and/or CXCR2 in eosinophils. In addition, we recently reported the functional expression of CXCR4. The ligand of CXCR4, stromal cell-derived factor-1 (SDF-1), was able to induce a strong migratory response comparable to that by eotaxin. In contrast to the CCR3/eotaxin system which is mainly regulated at the level of ligand production, the CXCR4/SDF-1 system is regulated at the level of receptor expression. CXCR4 expression was completely attenuated by IL-4 and IL-5 and upregulated by IFN-gamma and dexamethasone, while CCR3 expression was only marginally affected. The balance between the biological effects of these chemokine systems may affect the distribution and migration of eosinophils. PMID- 11408770 TI - Expression of VLA-4 on eosinophils decreases in patients with eosinophilia. AB - BACKGROUND: It is assumed that the very late antigen-4 (VLA-4) plays a key role in selective migration and accumulation of eosinophils to the allergic inflammatory focus. The regulatory mechanism for VLA-4 expression is poorly understood, as is its relationship between other adhesion molecules. OBJECTIVE: The aim of the study was to elucidate the relationship between VLA-4 expression and the activation of eosinophils. METHODS: The surface expression of VLA-4, Mac 1, ICAM-1, CD4, CD25, CD69, CD89, IL-5 receptor and GM-CSF receptor on eosinophils isolated from the peripheral blood of 15 patients with eosinophilia and 16 healthy volunteers was measured. RESULTS: The surface expression of VLA-4 presented in mean fluorescent intensity by flow-cytometric analysis showed a significant decrease in the patients with eosinophilia (>700 eosinophils/microl) compared to that of the subjects without eosinophilia. On the other hand, the surface expression of Mac-1 was significantly increased in the patients with eosinophilia. There was an inverse correlation between the expression of VLA-4 and that of Mac-1 (r = -0.81) on the eosinophils obtained from the patients with eosinophilia. CONCLUSION: The changes on the surface expressions of Mac-1 and VLA 4 may be indicating the activation of eosinophils in the patients with eosinophilia and may contribute to their migration to the allergic inflammatory focus. PMID- 11408771 TI - Effect of roxithromycin on eotaxin-primed reactive oxygen species from eosinophils. AB - BACKGROUND: The CC chemokine eotaxin not only attracts eosinophils to inflamed sites but also promotes adhesion, degranulation and reactive oxygen species production of eosinophils. Reactive oxygen species released from eosinophils are believed to injure epithelial cells at inflamed sites, resulting in airway hyperresponsiveness. Roxithromycin has been reported to have antiasthmatic effects, although its mechanism of action is not thoroughly understood. Therefore, the effect of roxithromycin on eotaxin-primed reactive oxygen species production from eosinophils was studied. METHODS: Reactive oxygen species production by eosinophils cultured with or without roxithromycin was evaluated using luminol-dependent chemiluminescence. RESULTS: Roxithromycin inhibited the release of reactive oxygen species from eosinophils evoked with the calcium ionophore A23187, regardless of pretreatment with or without eotaxin. CONCLUSION: Roxithromycin may protect epithelial cells at inflamed sites, at least partly by inhibiting the release of reactive oxygen species from eosinophils. PMID- 11408772 TI - A comparison of FcepsilonRI-mediated RANTES release from human platelets between allergic patients and healthy individuals. AB - BACKGROUND: Recently some studies have suggested that human platelets may play an important role in allergic inflammation through the high affinity IgE receptor (FcepsilonRI), the low affinity IgE receptor (FcepsilonRII/CD23) and the low affinity IgG receptor (FcgammaRIIA/CD32) expressed on the cell surface. We reported that human platelets via the FcepsilonRI induced the release of the chemical mediator serotonin and the chemokine RANTES (regulated upon activation, normal T expressed and presumably secreted), but the biological implication of human platelets in type I allergy has not yet been understood clearly. METHODS: We compared the levels of RANTES release from platelets obtained from allergic patients and healthy individuals, stimulated with monoclonal antibody (Ab) to human FcepsilonRI alpha-chain, or human myeloma IgE and anti-human IgE Ab. RESULTS: We confirmed that the level of RANTES release from platelets of allergic patients stimulated with human IgE and anti-human IgE was significantly higher than that of healthy individuals. CONCLUSIONS: We demonstrated that the surface expression levels of FcepsilonRI on the platelets from allergic patients and healthy individuals were not significantly different, but that the platelets of allergic patients were more activated by the IgE-FcepsilonRI pathway than those of healthy individuals. Taken together, these results suggest a novel and important role for human platelets in perpetuating allergic inflammation through the IgE and FcepsilonRI. PMID- 11408773 TI - IL-5 production by peripheral blood Th cells of adult asthma patients in response to Candida albicans allergen. AB - Cytokines produced by T cells are involved in chronic asthma. Relevant antigens for nonatopic asthma are still mostly unknown. Peripheral blood mononuclear cells (PBMCs) were obtained from adult asthmatic donors and incubated with Candida albicans extract. Proliferation response and cytokine production were analyzed. IL-2 and IL-5 were detectable in the cultures incubated with C. albicans extract, whereas IL-4 was undetectable. PBMCs obtained from some nonatopic asthma patients produced IL-5 upon stimulation of C. albicans extract. C. albicans allergen may be involved in the airway eosinophilic inflammation through the induction of IL-5 production by Th cells. PMID- 11408774 TI - A study of clinical features of cough variant asthma. AB - Patients with cough variant asthma (CVA) and classic asthma are frequently among subjects who present at clinics complaining of a chronic persistent cough. To reveal the features of CVA, we examined the differences in the clinical appearance between CVA and classic asthma. Ten CVA subjects and 11 classic asthmatics were enrolled in the study; they were recruited among patients who presented at the National Minamiokayama Hospital complaining of a chronic cough. The number of eosinophils in peripheral blood was 256 +/- 45.8/microl in CVA and 400 +/- 123/microl in classic asthma. Eosinophils represented 67% of the cells of sputum in CVA and 82% in classic asthma. Bronchial responsiveness to methacholine was Dmin 1.37 +/- 0.56 U in CVA and 0.71 +/- 0.46 U in classic asthma. There was no significant difference in these three parameters. There was only a significant difference in V25 between CVA and classic asthma, 80.0 +/- 6.9 and 52.2 +/- 10.0%, respectively. Eosinophil inflammation was almost the same in both CVA and classic asthma. PMID- 11408775 TI - Eosinophil infiltration and degranulation in normal human tissues: evidence for eosinophil degranulation in normal gastrointestinal tract. AB - Eosinophils play an important role in the pathogenesis of inflammatory diseases such as bronchial asthma and host immunity to parasitic infections. Deposition of eosinophil granule proteins and concomitant tissue damage have been documented in various diseases. Here, we review and summarize results of our immunofluorescence studies of eosinophil infiltration and degranulation in various normal human tissues. Furthermore, because eosinophil infiltration and degranulation are not normally present in healthy tissues, we examine whether eosinophil infiltration and degranulation normally occur in the small intestine and whether tissue procurement methods affect the extent of eosinophil infiltration and degranulation there. Hematopoietic and lymphatic tissues, including the thymus, showed eosinophil infiltration, but the only organ showing remarkable eosinophil infiltration and degranulation was the gastrointestinal (GI) tract. Eosinophil degranulation was significantly increased in specimens obtained by endoscopic forceps compared to those obtained by scalpel. These results suggest that tissue procurement methods affect the degree of eosinophil degranulation in the GI tract and that, among normal human body organs, both eosinophil infiltration and degranulation only occur in the GI tract. PMID- 11408776 TI - T cell vaccination eliminates antigen-specific T cells and prevents antigen induced eosinophil recruitment into the tissue. AB - Asthma is characterized by airway inflammation with prominent eosinophil infiltrates. In a murine model of asthma, antigen-induced eosinophil recruitment into the airways of sensitized mice is mediated by CD4+ T cells and their cytokines, especially IL-5. In the present study, using ovalbumin-specific T cell receptor transgenic mice, we found that T cell vaccination, which was the administration of preactivated and attenuated antigen-specific T cells by the intraperitoneal route, prevented antigen-induced eosinophil recruitment into the airways. This effect was antigen-specific because ovalbumin-specific T cell vaccination did not affect BSA-induced eosinophil recruitment into the airways. We also found that antigen-specific IgE production as well as antigen-induced proliferation and cytokine production of splenocytes were diminished by T cell vaccination. Moreover, flow-cytometric analyses revealed that T cell vaccination eliminated antigen-specific T cells in the periphery. Together, these results indicate that T cell vaccination prevents antigen-induced eosinophil recruitment into the airways presumably by eliminating antigen-specific T cells. PMID- 11408777 TI - Establishment and characterization of a murine mast cell line derived from NC/Nga mice. AB - A cell line, termed NCJ, was established from the bone marrow-derived mast cells (BMMCs) of NC/Nga mice that are mouse models for atopic dermatitis. NCJ cells expressed FcepsilonRI and c-kit and showed a metachromasia of the granules with a toluidine blue-positive and safranin-negative staining pattern that is characteristic for immature-type mast cells. Interestingly, NCJ cells showed proliferation independent of IL-3, which was associated with constitutive phosphorylation of Raf-1 and Erk kinases. Although NCJ cells had several characteristics of mast cells, we failed to detect FcepsilonRI-mediated beta hexosaminidase release and its histamine content. These findings indicated that NCJ cells represented a mast cell line with an immature phenotype and the ability to proliferate in the absence of mast cell growth factors. NCJ cells might thus be useful to study the molecular basis of mast cell proliferation. PMID- 11408778 TI - Effects of estradiol on glucoprivic transactivation of catecholaminergic neurons in the female rat caudal brainstem. AB - Hyperphagic and hypothalamic neuroendocrine responses to acute glucose deprivation are modified by the ovarian steroid estradiol (E). Observations of genomic activation of catecholaminergic (CA) neurons in the hindbrain lateral reticular nucleus, nucleus of the solitary tract, and area postrema (AP) by glucopenia support their potential function in pathways mediating regulatory effects of this metabolic challenge within the brain. Expression of E receptors by these cells suggests that their activity may be sensitive to steroid modulation during glucopenia. The present studies investigated the role of E on transcriptional activation of caudal brainstem CA neurons by the glucose antimetabolite, 2-deoxy-D-glucose (2DG). Ovariectomized rats were implanted with s.c. Silastic capsules containing E (30 or 250 microg/ml) or sesame oil, and injected i.p. 7 days later with 400 mg 2DG/kg or saline. Tyrosine hydroxylase (TH)-immunoreactive (-ir) neurons in the C(1)/A1, C2, C3, A2, A5, and A6 cell groups and AP were colabeled for Fos following antimetabolite administration, whereas vehicle injection resulted in negligible nuclear staining of these cells. With the exception of A2, A6, and AP cells, mean numbers of Fos- and TH-/Fos-ir positive neurons in these brain sites did not differ between E- and sesame oil implanted groups. Numbers of TH-positive A2 and A6 neurons that expressed Fos in response to 2DG were significantly greater in rats implanted with the high E dose vs. either the low steroid dose or sesame oil. These results show that the magnitude of cellular Fos labeling within discrete hindbrain CA neuron populations varies in accordance with circulating E levels. These findings suggest that E may exert potential modulatory effects on glucoprivic activation of the Fos stimulus/transcription cascade and consequent compensatory genomic responses within specific areas of the female rat caudal brainstem. PMID- 11408779 TI - Progesterone treatment that either blocks or augments the estradiol-induced gonadotropin-releasing hormone surge is associated with different patterns of hypothalamic neural activation. AB - Progesterone can either augment or inhibit the surge of gonadotropin-releasing hormone (GnRH) that drives the preovulatory luteinizing hormone (LH) surge. This study investigated the central mechanisms through which progesterone might achieve these divergent effects by examining the effects of exogenous steroids on the activation of GnRH neurons and non-GnRH-immunopositive cells in the preoptic area/anterior hypothalamus of steroid-treated ovariectomized ewes. Fos expression (an index of cellular activation) was examined during the estradiol-induced GnRH surge in ewes treated with progesterone using regimes that have been reported to either augment (progesterone pretreatment) or inhibit (progesterone treatment at the time of the surge-inducing estradiol increment) the GnRH surge. Control groups received either no progesterone pretreatment or no surge-inducing estradiol increment. Induction of an LH surge was associated with a significant (p < 0.0001) increase in the proportion of activated GnRH neurons, irrespective of whether ewes received progesterone pretreatment. However, the number of non GnRH-immunopositive cells activated during the surge was significantly (p < 0.0001) increased in ewes that received the progesterone pretreatment. By contrast, the proportion of GnRH neurons and non-GnRH-immunopositive cells that expressed Fos was significantly (p < 0.0001) reduced in ewes in which the surge was inhibited by progesterone compared to ewes in which a surge was stimulated. These data indicate that (1) progesterone pretreatment increases the activation of non-GnRH cells during the estradiol-induced surge, but does not affect the proportion of GnRH neurons activated and (2) when administered concurrently with a surge-inducing estradiol increment, progesterone prevents the activation of GnRH neurons and non-GnRH cells that is normally associated with the estradiol induced surge. Therefore, progesterone does not appear to augment the GnRH surge by increasing the proportion of GnRH neurons that are activated by estradiol, whereas inhibition of the GnRH surge involves prevention of the activation of GnRH neurons. Thus, the augmentation and inhibition of the GnRH surge by progesterone appear to be regulated via different effects on the GnRH neurosecretory system. PMID- 11408780 TI - Protooncogene c-fos Involvement in the Molecular Mechanism of Rat Brain Sexual Differentiation. AB - Brain sexual differentiation is mediated through testosterone, which acts during the perinatal period in the form of both 5alpha-dihydrotestosterone and estradiol. In order to gain insight into the molecular mechanisms involved, we studied induction of c-fos, an index of functional neuronal activation, in the 2 day-old female rat brain after injection of a masculinizing dose of testosterone. Administration of testosterone resulted in induction of c-fos gene expression in the hypothalamus, as determined by Northern analysis. Following immunocytochemistry, we demonstrated an increase in the number of Fos-positive nuclei in the median and medial preoptic nucleus, the medial preoptic area extending to the lateral preoptic area, and the peri- and paraventricular area. In an effort to see whether testosterone acted as 5alpha-dihydrotestosterone or as estradiol, we injected either steroid and looked at fos induction. Estradiol mimicked the effect of testosterone, while 5alpha-dihydrotestosterone was without effect. Furthermore, injection of an estrogen receptor blocker, clomiphene, together with testosterone, abolished the testosterone-induced increase in Fos positive nuclei, thus confirming the finding that testosterone induces c-fos by acting through estrogen receptors. Electrophoretic mobility shift assays showed that nuclear extracts from 2-day-old female hypothalamus contain a protein, most probably the estrogen receptor, which binds specifically to oligodeoxynucleotides with the sequence of either vitERE, the consensus estrogen-responsive element (ERE) found in the vittelogenin gene, or fosERE, the ERE found in the 3' untranslated region of the mouse c-fos gene. This suggests that the effect of testosterone-derived estradiol on c-fos expression is a direct one, mediated by binding of estrogen receptors to an ERE in the c-fos gene-regulatory regions. PMID- 11408781 TI - Sensitivity to a neurosteroid is increased during addition of progestagen to postmenopausal hormone replacement therapy. AB - The aim of this study was to compare the pharmacodynamic response to a neuroactive steroid, pregnanolone, before and during different hormonal settings of postmenopausal hormone replacement therapy (HRT). Twenty-seven postmenopausal women with climacteric symptoms were administered HRT in a randomized, double blinded, placebo-controlled crossover study. The women received 2 mg estradiol (E2) continuously during four 28-day cycles and 10 mg medroxyprogesterone acetate (MPA), 1 mg norethisterone acetate (NETA) or placebo sequentially for the last 14 days in each cycle. The pharmacodynamic response to pregnanolone was assessed before treatment and during the last week of each treatment, by comparing the effects of intravenous pregnanolone (3alpha-hydroxy-5beta-pregnan-20-one) on saccadic eye velocity (SEV), saccade deceleration, saccade latency and self-rated sedation. Throughout the study daily symptom rating scales were kept. During the progesta gen phase of the treatment cycles, negative mood symptoms and physical symptoms were increased, whereas positive mood symptoms were decreased. Compared to pretretreatment conditions, E2 alone did not change the responsiveness to pregnanolone. During progestagen addition to E2, the responsiveness to pregnanolone was increased. The sedation response increased compared to pretreatment conditions during both E2 + MPA and E2 + NETA treatment. Compared to E2 treatment alone, addition of MPA increased the postpregnanolone effect on saccade deceleration, whereas the SEV response to pregnanolone was increased during E2 + NETA treatment. It is concluded that pregnanolone sensitivity increases together with deterioration in mood symptoms during addition of progestagen to HRT. PMID- 11408782 TI - Release of luteinizing hormone from the anterior pituitary gland in vitro can be concurrently regulated by at least three peptides: gonadotropin-releasing hormone, oxytocin and neuropeptide Y. AB - Normal action of the female ovulatory cycle is dependent on a surge of luteinizing hormone (LH) from the pituitary. Regulation of the levels of LH is therefore vital to reproductive function. It has long been established that gonadotropin-releasing hormone (GnRH) is an important component of the regulatory processes. Other peptides, including oxytocin and neuropeptide Y (NPY), have also been observed to affect LH activities. However, the possibility of the concurrent actions of these peptides has rarely been considered despite their documented presence in the hypophyseal blood during pro-oestrus. In this study, the direct effects of oxytocin and NPY on LH release were studied, as well as the effects of both peptides simultaneously. Also, pituitaries were stimulated with GnRH, and the effects of pre-exposure of the pituitary tissue to oxytocin or NPY were investigated. Further, the effect of oxytocin and NPY together on GnRH stimulation of LH release was determined. Anterior pituitaries were collected from adult female rats on the morning of pro-oestrus. Hemipituitaries were cut in two and placed in a chamber of a perifusion system. The pituitary tissue was perifused with medium alone, oxytocin and NPY, separately or in combination, for 2 h after an initial 100-min equilibration period with no peptide present. Fractions of eluate were collected and LH was measured by radioimmunoassay. LH output was stimulated by both oxytocin (p < 0.01) and NPY (p < 0.02). Furthermore, the addition of NPY to oxytocin during the perifusion elicited a further increase in LH release (p < 0.05). The pituitary tissue was exposed to a 4-min pulse of GnRH after 220 min. Stimulation of LH release by GnRH was synergistically augmented by exposure of the tissue to either oxytocin (p < 0.01) or NPY (p < 0.05) for the immediately preceding 2 h. When NPY was added to oxytocin in the perifusion medium, stimulation of LH release by GnRH was increased even further (p < 0.05). Oxytocin also synergistically enhanced the effect of a second, primed GnRH pulse, whereas the effect of NPY was less robust. This study therefore demonstrated that LH release is modified in the presence of oxytocin, NPY or GnRH alone, and also that combinations of the peptides produce further variations in LH output. Therefore, the concentrations of LH that are present in vivo are likely to be at least partly the result of the co-ordinated effects of combinations of peptides acting on the pituitary. PMID- 11408783 TI - Somatostatin inhibits alpha-2-adrenergic-induced secretion of growth hormone releasing hormone. AB - The purpose of this experiment was to determine the role of growth hormone releasing hormone (GHRH) and somatostatin (SRIH) neurons in mediating alpha(2) adrenergic receptor-induced stimulation of growth hormone (GH) secretion in cattle. Our first objective was to determine if stimulation of alpha(2) adrenergic receptors increases activity of GHRH neurons in the arcuate nucleus (ARC) and/or decreases activity of SRIH neurons in periventricular (PeVN) and ARC nuclei. Clonidine (an alpha(2)-adrenergic agonist) or vehicle (saline) were injected i.v. into steers and dual-label immunohistochemistry was performed to quantify the number of GHRH and SRIH neurons expressing Fos and Fos-related antigens (Fos/FRA) as markers of neuronal activity. Clonidine increased concentrations of GH in serum and decreased activity of SRIH neurons in the PeVN, but not in the ARC. Clonidine did not alter activity of GHRH neurons in the ARC. Our second objective was to determine if clonidine decreases secretion of SRIH from perifused slices of hypothalami, which contain perikarya and terminals of GHRH and SRIH neurons, and from explants of hypophysial stalk alone, which contain only terminals of GHRH and SRIH neurons. Clonidine failed to alter release of GHRH or SRIH from hypothalamic slices, but stimulated release of GHRH from explants of hypophysial stalk. Blockade of SRIH receptors enabled clonidine to stimulate release of GHRH from slices of hypothalami, but also stimulated release of SRIH. These results suggest that alpha(2)-adrenergic-induced secretion of GH occurs via a dual mechanism involving inhibition of SRIH neurons in the PeVN and direct stimulation of GHRH release from axon terminals in the median eminence. PMID- 11408784 TI - Amphibian melanotrope subpopulations respond differentially to hypothalamic secreto-inhibitors. AB - The melanotrope population of the frog intermediate lobe consists of two subtypes of cells, referred to as high-(HD) and low-density (LD) melanotrope cells, which differ markedly in their basal morphofunctional features as well as their in vitro response to hypothalamic factors, such as the stimulator thyrotropin releasing hormone (TRH) and the inhibitor dopamine. In this study, we have investigated whether other major hypothalamic regulators of the release of alpha melanocyte-stimulating hormone (alpha-MSH), such as gamma-aminobutyric acid (GABA) and neuropeptide Y (NPY), also differentially regulate frog melanotrope subpopulations. Our results show that in LD cells, both factors markedly inhibited proopiomelanocortin (POMC) mRNA accumulation and alpha-MSH secretion. In contrast, the secretory and biosynthetic activity of HD cells was not modified by GABA. NPY inhibited POMC transcript accumulation and tended to reduce alpha MSH secretion in HD cells, yet these effects were less pronounced than those evoked in LD cells. In addition, GABA and NPY inhibited the KCl-induced rise in cytosolic free calcium levels in both subpopulations. Taken together, these results further indicate that frog melanotrope subpopulations are differentially regulated by the hypothalamus and strongly suggest that the intensity of such regulation is directly related to the activity of the cell subset. Thus, the LD subpopulation represents a highly responsive cell subset which is regulated by multiple neuroendocrine factors (TRH, dopamine, GABA and NPY), whereas the hormone storage HD subpopulation shows a moderate response to single stimulatory (TRH) and inhibitory (NPY) inputs. PMID- 11408785 TI - Levels of monocyte reactive oxygen species are associated with reduced natural killer cell activity in major depressive disorder. AB - Major depressive disorder (MDD) is associated with reductions in natural killer cell activity (NKCA), however the mechanism(s) mediating reduced NKCA in MDD has yet to be determined. In light of evidence that MDD is associated with an inflammatory immune response, we propose that reactive oxygen species (ROS), generated by inflammatory leukocytes (monocytes and/or neutrophils), may mediate the suppression of NKCA in MDD. Intracellular levels of monocyte ROS were significantly associated with reductions in NKCA in outpatients (n = 15) diagnosed with MDD. Sleep disturbance was also significantly correlated with reductions in NKCA. Elevated levels of ROS may be an additional characteristic of a subset of depressed patients in whom an inflammatory response persists and elevations in ROS may, in part, mediate the associations observed between MDD, cardiovascular disease, and cancer. PMID- 11408786 TI - Contingent negative variation and personality in depression. AB - Contingent negative variation (CNV) has been applied in depression with controversial results. A major source for these controversial results could result from the diversity of depressed patients included in the different studies. Supporting this assumption, impulsivity, blunted affect and suicidal behavior significantly influence CNV amplitude. However, no data are available on the possible influences of personality of depressed patients on CNV. Since personality is related to CNV in normal subjects, the aim of the present study was to investigate the relationship between CNV and personality as assessed by the Temperament and Character Inventory (TCI) in 52 depressed patients. A group of 76 healthy volunteers was included in the study. Among depressed patients, the main results of the study shows that CNV amplitude is positively correlated with self-transcendence dimension. In healthy volunteers, results show that CNV amplitude is negatively correlated with novelty seeking and persistence dimensions. The other dimensions are not related to CNV amplitude either in depressed patients or in healthy volunteers. The findings among depressed subjects suggest that lower CNV amplitude may be associated with psychotic traits (high self-transcendence scores), and those observed in healthy subjects are consistent with previous studies and support energetical models of slow potentials. However, the preliminary nature of the present results with respect to the weak statistical significance should be underlined. PMID- 11408787 TI - Cerebrospinal fluid 5-hydroxytryptamine and 5-hydroxyindoleacetic acid in HIV-1 infection. AB - Reduced level of serotonin (5-hydroxytryptamine, 5-HT) in humans has been associated with a number of mental health and behavioral problems including depression, aggression, violence, sexual dysfunctions, sleep and eating disorders. Even though among HIV-1-infected individuals, prevalence of mental health and behavioral problems are common, their relationship with central nervous system serotonin functions is not clearly understood. This investigation was carried out to study the status of CSF 5-HT in HIV-1+ subjects (n = 21), in the early stage of infection, and HIV-1- control subjects (n = 24). Samples of CSF were obtained by lumbar puncture and were analyzed for 5-HT and its metabolite 5-hydroxyindoleacetic acid (5-HIAA), using high-performance liquid chromatography equipped with electrochemical detector. Levels of CSF 5-HT were significantly lower in the HIV-1+ group compared to the HIV-1- group. There was no significant difference in the CSF 5-HIAA levels between the two groups. In both groups, however, there was a significant correlation between CSF 5-HT and 5 HIAA. In the HIV-1 + group, although CSF 5-HT level was significantly negatively correlated with serostatus, there was no correlation between either CSF 5-HT or 5 HIAA levels and CD4 cell number or any behavioral measures evaluated in this study, including Beck's Depression Inventory and state/trait anxiety scores. These data suggest that HIV-1 infection affects the CNS 5-HT status with no significant association with measures of depression and anxiety, at least in the early stage of infection. PMID- 11408788 TI - Season of birth variations in the temperament and character inventory of personality in a general population. AB - BACKGROUND: Since several studies show season of birth variations in morbidity, suicidal behavior and CSF (cerebrospinal fluid) monoamine metabolites, we investigated season of birth variations in personality in the population. METHODS: We analyzed by multiple logistic regressions the Temperament and Character Inventory (TCI) for 2,130 individuals taking part in the Betula prospective random cohort study of Umea, Sweden. RESULTS: The personality dimensions were correlated significantly with age and gender. We stratified the data according to age, gender and the season of TCI measurement. By the median split in each stratum, a high-value group and a low-value group were obtained for each of the personality dimensions. Those born during February to April were significantly more likely than those born during October to January to have high NS (novelty seeking) among women, particularly the subscale NS2 (impulsiveness vs. reflection), and to have high PS (persistence) among men. Temperament profiles also showed season of birth variations. CONCLUSIONS: We discuss the associations in the literature between personality and the monoamines serotonin and dopamine, and suggest that our results are compatible with a hypothesis of season of birth variation in the monoamine turnover. The personality traits are likely to be influenced by several genetic and environmental factors, one of them being the season of birth. PMID- 11408789 TI - Association study for a functional serotonin transporter gene polymorphism and late-onset Alzheimer's disease for Chinese patients. AB - Two recent studies have demonstrated an association for a deletion/insertion polymorphism within the promoter region of the serotonin transporter gene (5 HTTLPR), and Alzheimer's disease (AD). According to these studies, subjects with the short variant of the 5-HTTLPR gene are at increased risk for AD; however, this finding has not been confirmed by other workers. To evaluate the role of the 5-HTTLPR gene in susceptibility for AD, we conducted an association study for this polymorphism in a Chinese population. No significant differences were determined for genotype distribution or allele frequencies, comparing AD patients and normal controls. Even dividing the population into subgroups according to the presence of the APOE epsilon4 allele, no differences for genotype or allele frequencies were determined, comparing patients and controls. These results suggest that it is unlikely that the 5-HTTLPR polymorphism plays a substantial role in conferring susceptibility to AD. PMID- 11408790 TI - Association study of angiotensin-converting enzyme gene polymorphism with schizophrenia and polydipsia. AB - Angiotensin-converting enzyme (ACE) is a key enzyme in the renin-angiotensin system and can modulate dopamine turnover in the midbrain. Previous studies have revealed changes in the central ACE levels for schizophrenic patients, possibly related to the polydipsia commonly demonstrated for chronic schizophrenia. An insertion (I)/deletion (D) polymorphism of the ACE gene has been associated with ACE levels. Therefore, we elected to investigate the ACE I/D polymorphism for 124 schizophrenic patients and 117 control subjects. No significant differences for the genotype distribution or the allele frequency were revealed comparing controls and schizophrenic patients. The ACE genotypes were not associated with onset age or psychiatric symptoms for the schizophrenic cases. A modest association was revealed for this ACE polymorphism and polydipsia diagnosis for these patients. Using bearers of the D allele as baseline, the ratio for I/I homozygote was 2.31 (95% CI 0.95-5.65). This association needs further replication as it may have implications for the pathogenesis and the treatment of polydipsia for schizophrenic patients. PMID- 11408791 TI - Reduced regional [14C]2-deoxyglucose uptake in response to long-term clozapine administration in rats. AB - Clozapine has superior effects in treating negative symptoms of schizophrenia and causes less extrapyramidal side effects than traditional antipsychotics. In this study, we investigated the effects of acute and long-term clozapine administration on [14C]2-deoxyglucose uptake (2-DG uptake) in rats, as measured using the [14C]2-deoxy-D-glucose method. The 2-DG uptake was reduced in fewer regions after chronic clozapine (46%) than after acute clozapine (97%). After chronic clozapine treatment, the 2-DG uptake was reduced in the shell, but not the core, of the nucleus accumbens. In addition, long-term clozapine treatment remained affecting 2-DG uptake in several regions of the extrapyramidal system and the thalamus. The pattern of 2-DG uptake changes after long-term clozapine administration may provide information for the regions related to the therapeutic effect of clozapine. PMID- 11408792 TI - Consensus on the Practical Use of Amisulpride, an Atypical Antipsychotic, in the Treatment of Schizophrenia. AB - Clinical expectations in schizophrenia treatment have greatly increased since the introduction of new atypical antipsychotics, but the choice of therapeutic strategy has become more complex and reference guidelines are scarce. This paper summarizes the consensus of a broad range of professionals after long-term commercialization in France of an atypical antipsychotic, amisulpride. Participants were from psychiatric hospitals, private clinics, out-patients settings and research; all were experienced with the drug. Discussions focused on the practical use of amisulpride, as, in addition to the double-blind trials information, it may be useful for psychiatrists of other countries to intuitively understand the therapeutic properties of amisulpride. The topics selected include acute psychotic episodes, short- and long-term follow-up, feasibility of treating the initial phase, the elderly and switching treatments. The French experience emphasizes the central role of amisulpride as a first-line treatment of schizophrenia. PMID- 11408793 TI - The influence of personality on nicotine craving: a hierarchical multivariate statistical prediction model. AB - The present study proposes a hierarchical multivariate statistical prediction model which enables to determine the most prominent variables (physiological, biochemical and personality factors) related to nicotine craving and dopaminergic activation. Based on animal studies reporting a reduction of the rewarding effects of psychotropic drugs after blockade or destruction of the mesolimbic dopamine (DA) system, changes in nicotine craving after pharmacological manipulation by means of a DA agonist (lisuride 0.2 mg) and a DA antagonist (fluphenazine 2 mg) were assessed in 36 healthy male heavy smokers. The major aim was the development of a multivariate prediction model which is applicable in samples lacking variance homogeneity or the prerequisite of a multivariate normal distribution. The model proposed is a combination of multivariate parametric and nonparametric methods taking advantage of their individual merits. Especially personality variables, such as sensation seeking, impulsivity, and neuroticism showed to be important predictors of craving in this responder approach. PMID- 11408794 TI - EEG effects of smoking: Is there tachyphylaxis? AB - Smoking generally affects the human EEG by reducing low-frequency activity and increasing higher-frequency activity. Using a double-blind design, we sought to determine if acute tolerance (tachyphylaxis) to this effect can be observed. EEG was recorded in overnight-abstaining participants before and after smoking three cigarettes at 40-min intervals in two separate sessions. In one session ('short interval'), all three cigarettes had 'typical' nicotine yields (1.1 mg, FTC method). In the other session ('long interval'), the second cigarette had a very low nicotine yield (0.05 mg; thus making the interval between 1.1-mg cigarettes 80 min). Eyes-closed alpha power decreased and eyes-open beta-2 power increased following smoking each of the 'typical-yield' cigarettes but not the low-yield cigarettes. The decrease in alpha power after smoking the 'typical-yield' cigarette at the 40- and 80-min intervals was less than that following the first cigarette of the day, indicating tachyphylaxis. In contrast, the increase in eyes open beta-2 power did not differ among cigarettes regardless of the order or interval between 'typical-yield' cigarettes. EEG changes in other frequency bands produced by smoking the first cigarette of the day were not consistent across sessions, making interpretation in terms of tachyphylaxis somewhat problematic. Overall, lower-frequency alpha activity displayed patterns consistent with tachyphylaxis while beta-2 did not. PMID- 11408795 TI - The role of Epstein-Barr virus in neoplastic transformation. AB - In this review, we focus on new data from basic, translational and clinical research relating to the Epstein-Barr virus (EBV). Beside its well-known tropism for B lymphocytes and epithelial cells, EBV also infects T lymphocytes, monocytes and granulocytes. After primary infection, EBV persists throughout the life span in resting memory B cells, from where it is reactivated upon breakdown of cellular immunity. In the process of neoplastic transformation, the EBV-encoded latent membrane protein 1 (LMP1) oncogene represents the major driving force. LMP1 acts like a constitutively activated receptor of the tumor necrosis factor receptor family and allows the amplification or bypassing of physiological regulatory signals through direct and indirect interactions with proteins of the tumor necrosis factor receptor-associated factor (TRAF) family. TRAF2-mediated NF kappaB activation, AP-1 induction and JAK3/STAT activation may result in sustained proliferation leading to lymphoma. The ability of LMP1 to suppress germinal center formation and its capacity to mediate its own transcriptional activation shed new light on the pathogenesis of EBV-associated latency type II lymphoproliferations like Hodgkin's disease and angioimmunoblastic lymphadenopathy. The carboxy terminus of LMP1 is also a reliable marker for individual EBV strain identification and thus offers new possibilities in tracing the molecular events leading to posttransplant lymphoproliferative disorders (PTLDs). Cytotoxic T lymphocytes directed against well-characterized epitopes of EBV latency genes represent an already successful and promising therapeutic approach to EBV-associated lymphomas, in particular PTLDs. PMID- 11408796 TI - Single-agent gemcitabine is active in previously treated metastatic breast cancer. AB - BACKGROUND: This phase II study was designed to assess the efficacy and safety of gemcitabine in patients with metastatic breast cancer (MBC) previously treated with an anthracycline- or anthracenedione-containing regimen as first-line therapy for metastatic disease. PATIENTS AND METHODS: Forty-seven patients with MBC were enrolled in five French centers. Patients were eligible if they had received one prior chemotherapy regimen with an anthracycline or anthracenedione for metastatic disease, if they had responded to that prior regimen, and if they had relapsed at least 6 months after the first response. Fifteen patients received more than one prior anthracycline regimen; thus, gemcitabine was third line therapy for 30% of patients. Gemcitabine 1,200 mg/m(2) was administered as a 30-min intravenous infusion on days 1, 8, and 15 of a 28-day cycle for a maximum of eight cycles after the best response was obtained. RESULTS: Objective responses were seen in 12 of 41 assessable patients (4 complete responses and 8 partial responses), for an objective response rate of 29% (95% confidence interval, 16-46%). The median response duration was 8.1 months (range: 2.5-27.4 months). Serious hematological toxicity was minimal, with grade 4 neutropenia in 2% of the patients (no neutropenic fever), grade 3 neutropenia in 28% of the patients, and grade 3 thrombocytopenia in 6% of the patients. Among the nonhematological toxicities, asthenia was the most common. CONCLUSIONS: Gemcitabine given at this dose and schedule is a well-tolerated treatment with definitive antitumor activity in pretreated MBC patients. This result warrants future exploration of the use of gemcitabine as a single agent and in combination in patients with MBC. PMID- 11408797 TI - Progress report on the palliative therapy of 100 patients with neoplastic effusions by intracavitary low-dose interleukin-2. AB - OBJECTIVE: Several cytokines, particularly IL-2 and interferons, are thought to be effective in the palliative therapy of neoplastic effusions. We report on the activity and toxicity of intracavitary administration of low-dose IL-2 in a case series of 100 cancer patients with neoplastic effusions. METHODS: One hundred patients with advanced solid tumors and neoplastic effusions underwent IL-2 intracavitary injection as first-line treatment. The most common sites of fluid accumulation were pleura (n = 68), peritoneum (n = 21) and pericardium (n = 11). Breast cancer, lung cancer and mesothelioma were the most frequent neoplasms in our series. One cycle consisted of intracavitary IL-2 at 6,000,000 IU on days 1 and 7. RESULTS: According to Paladine's criteria, an objective clinical response was achieved in 72% (complete response in 27% and partial response in 45%), with a median duration of 5 months (range: 1-11 months). The peritoneum was the least responsive site for neoplastic effusion reduction. IL-2 intracavitary injection was well tolerated in all patients; the only toxicity observed was fever >38 degrees C in 6% of the patients. CONCLUSION: This study shows that intracavitary injection of IL-2 represents a feasible, well-tolerated and effective therapy of neoplastic fluid accumulation. Further studies are needed in order to compare the effectiveness of intracavitary IL-2 with other standard treatments. PMID- 11408798 TI - Treatment of advanced hepatocellular carcinoma with biweekly high-dose gemcitabine. AB - INTRODUCTION: Until today, an optimal palliative treatment regimen has not been defined for patients with advanced hepatocellular carcinoma. Since the novel cytidine analog gemcitabine has shown strong antitumor effects in vitro in a human hepatoma cell line and its therapeutic potential seems well established in several different tumors including gastrointestinal adenocarcinomas, the present phase II trial using a dose-intensified biweekly administration schedule was initiated. PATIENTS AND METHODS: 17 patients with histologically confirmed unresectable advanced or metastatic hepatoma were treated with gemcitabine 2,200 mg/m(2) given as a 30-min intravenous infusion on days 1 and 15. Treatment courses were repeated every 4 weeks. RESULTS: All patients were evaluable for response and toxicity assessment. No objective response was achieved, stable disease occurred in 8 patients (47%), and 9 progressed while on chemotherapy. The median time to progression was 4 months (range 1.5-14 months), and the median survival time was 8.5 months (range 2.5-16.0+ months). Treatment was well tolerated with mild or moderate leukopenia, thrombocytopenia and anemia representing the most common side effects. Gastrointestinal and other subjective toxicities were infrequent and also generally mild. CONCLUSIONS: In view of the disappointing treatment results, gemcitabine using this particular dose regimen should not be considered for further investigation in patients with advanced hepatocellular carcinoma. PMID- 11408799 TI - Combination gemcitabine and docetaxel therapy in advanced adenocarcinoma of the pancreas. AB - OBJECTIVE: To determine the clinical and laboratory response rate of a gemcitabine and docetaxel combination in human adenocarcinoma of the pancreas in vitro and in vivo. METHODS: Fifteen patients with unresectable pancreatic cancer were treated with gemcitabine, 900 mg/m(2), and docetaxel, 90 mg/m(2), every 3 weeks. Two human pancreatic cancer lines were tested in MTT assays for their response to titrations of gemcitabine and/or docetaxel at different time points and scheduling for biochemical synergy or additional antitumor effects. RESULTS: In vitro testing showed that these two agents were minimally effective alone but when combined, they displayed additional biochemical antiproliferative effects in MTT assays. With intent-to-treat analysis of all 15 patients, 4 patients (27%) achieved an objective response by CT scan, including one complete response. Seven patients (47%) had subjective improvement and decreased serum marker levels of CA 19-9. None of the 12 patients without prior therapy developed nadir white blood cell counts below 1,000/mm(3); 2 of 3 patients with prior radiation therapy developed nadir white blood cell counts below 1,000/mm(3). CONCLUSION: This regimen is well tolerated and appears to have a significant objective response rate. Gemcitabine and docetaxel antitumor effects are additive in vitro, which may help to explain the response rate. PMID- 11408800 TI - Primary pancreatic lymphoma: clinicopathological analysis of 19 cases from Japan and review of the literature. AB - Primary pancreatic lymphoma (PL) is an extremely rare disease, and large-scale studies are rarely performed even in Western countries, in which all cases of PL reported to date were of the B-cell type. Little information regarding PL is available in Japan. Nineteen cases of PL were collected through a nationwide study in Japan, and paraffin-embedded specimens were processed for staining with hematoxylin and eosin and by immunohistochemical procedures. Clinicopathological findings were reviewed and compared with those reported in Western countries. The patient population consisted of 13 men and 6 women, ranging in age from 46 to 84 (average 62) years. Abdominal pain was the most common presenting symptom. Tumors were located in the pancreatic head (12 cases), tail (4 cases) and body (2 cases), and ranged in size from 4 to 17 cm. Clinical stage was I(E) in 9 cases and II(E) in 10. Radical resection was performed in 10 cases and bypass operation in 1, followed by chemotherapy in 8. Immunophenotypically, 15 cases were B-cell and 4 were T-cell lymphomas. Seventy-three percent of B-cell tumors were diffuse large B-cell lymphomas. The 1-year actuarial survival rate for B-cell lymphomas (51.9%) was better than that of T-cell lymphomas (0%). However, in Japan the incidence of T-cell PLs was higher, and, partly as a consequence of this, prognosis was poorer than in Western countries. PMID- 11408801 TI - Interferon alpha induction of Stat1 and Stat2 and their prognostic significance in carcinoid tumors. AB - IFN-alpha has been shown to elicit antitumor activity in carcinoid tumors. In the present study the effects of IFN-alpha on the protein expression involved in the IFN-alpha signaling, which were recognized as signal transducer and activators of transcription (Stats), were investigated. Archive specimens from 45 carcinoid patients, 33 before IFN-alpha and 45 during treatment, were studied. The tissues were immunostained for protein expression by using monoclonal anti-Stat1 and anti Stat2 antibodies. Results showed that Stat1 and Stat2 immunostaining were significantly increased during IFN-alpha treatment. Stats scores were significantly increased only in patients with objective response as well as those with stable disease but not in those with progressive disease. The induction of Stats correlated with the survival rates of the patients. In addition, both Stat scores significantly correlated with the p68 protein expression. In a carcinoid tumor cell line, Bon1, IFN-alpha dose-dependently increased the Stat expression. Analysis of cell fractions showed that IFN-alpha increased Stat protein both in cytoplasm and nucleoplasm of the cells. Furthermore, IFN-alpha enhanced tyrosine phosphorylation of Stat1 and Stat2. Thus, the antitumor effect, in vivo and in vitro, in IFN-alpha-treated carcinoid tumors seems to be mediated via upregulation of Stat proteins and enhancement of phosphorylation of these proteins. PMID- 11408802 TI - Immunohistochemical evaluation of cadherin and catenin expression in early gastric carcinomas: correlation with clinicopathologic characteristics and Helicobacter pylori infection. AB - Dysfunction of E-cadherin and catenin has been linked to invasiveness and differentiation of tumors. This study aimed to characterize the expression of cadherins and catenins in early gastric carcinoma and their relationship to clinicopathologic characteristics and Helicobacter pylori infection. E-cadherin and alpha-, beta- and gamma-catenins were strongly expressed in normal epithelium but abnormal immunoreactivity of at least one of these four proteins was noted in 48 (90.6%) of 53 early gastric carcinomas. Only 5 cases with intestinal-type tumors had intact expression of E-cadherin and alpha-, beta-, and gamma-catenins. Abnormal immunoreactivity in the tumor tissue was observed in 18 patients (34.0%) for E-cadherin, in 35 (66.0%) for alpha-catenin, in 20 (37.7%) for beta-catenin, and in 37 (69.8%) for gamma-catenin. In diffuse-type tumors, abnormal expression of E-cadherin (60.9 vs. 13.3%, p < 0.0005), alpha-catenin (82.6 vs. 53.3%, p < 0.05) and gamma-catenin (91.3 vs. 53.3%, p < 0.005) was more frequent than in the intestinal type. Ten tumors with lymph node metastasis showed a relatively higher frequency of abnormal expression of E-cadherin (70 vs. 25.6%, p < 0.05) but a lower frequency of abnormal expression of beta-catenin (10 vs. 44.1%, p = 0.07) than those without metastasis. No significant association was found between cadherin/catenin expression and the depth of invasion or the H. pylori status. It was concluded that abnormal expression of E-cadherin and the catenin-mediated cell-cell adhesion system occurs frequently in early gastric carcinogenesis and may play an important role in the genesis of histologic differentiation and in the mode of metastasis of early gastric carcinomas. PMID- 11408803 TI - Disruption of the p16/cyclin D1/retinoblastoma protein pathway in the majority of human hepatocellular carcinomas. AB - p16, cyclin D1 and retinoblastoma protein (pRB) regulate G1 to S transition and are commonly targeted in various cancers. However, few studies have simultaneously examined all components of the p16/cyclin D1/pRB pathway (RB pathway) in hepatocellular carcinoma (HCC). To clarify the role of the disruption of the RB pathway in HCC, we analyzed p16, pRB and cyclin D1 in 47 HCCs. Inactivation of p16 was detected in 30 of 47 HCCs (64%) by Western blot analysis and significantly correlated with hypermethylation of the promoter of this gene. pRB expression was found to be absent in 13 of 47 HCCs (28%) by immunohistochemistry. We found that 38 of 47 HCCs (81%) contained at least one inactivation in either pRB or p16. Furthermore, there was a significant inverse correlation between p16 and pRB inactivation (p = 0.041). Overexpression of cyclin D1 was detected in 5 of 47 HCCs (11%) by immunohistochemistry. The cases with cyclin D1 overexpression exhibited an advanced clinicopathological appearance and also contained inactivation of pRB and/or p16. These findings suggest that inactivation of pRB and/or p16 is a major event in human hepatocarcinogenesis, while cyclin D1 overexpression may confer additional growth advantages to the tumor in addition to pRB and/or p16 inactivation in HCC. PMID- 11408804 TI - Estrogens, testosterone and sex hormone binding globulin in relation to liver cancer in men. AB - OBJECTIVE: Liver disease in men has been associated with an imbalance of serum estradiol and testosterone. We have evaluated whether serum estradiol and testosterone levels are altered in male liver cancer patients as a result of a specific effect of the disease or because of the associated liver damage. METHODS: We have performed a hospital-based case-control study in Greece. The study subjects were all men; 73 patients with hepatocellular carcinoma (HCC), 25 with metastatic liver cancer (MLC) patients and 111 control subjects. Serum estradiol, testosterone and sex hormone binding globulin (SHBG) levels were measured for each subject. Data were analyzed by multiple linear regression. RESULTS: Mean serum estradiol levels were significantly higher among HCC patients as well as among patients with MLC compared to controls. Mean serum testosterone levels were significantly lower among HCC patients as well as among patients with MLC compared to controls. The mean SHBG levels did not differ significantly between the groups. After controlling for the degree of liver damage, the elevated serum estradiol and reduced serum testosterone levels among HCC and MLC patients were no longer significant. CONCLUSIONS: Changes in sex steroid levels among patients with liver damage are due to the liver damage per se and not to specific disease processes. PMID- 11408805 TI - Loss of ErbB2 expression in pulmonary metastatic lesions in osteosarcoma. AB - OBJECTIVES: The c-erbB2 protooncogene is located on human chromosome 17 and encodes a 185-kilodalton transmembrane glycoprotein (ErbB2). Elevated ErbB2 expression or gene amplification has been shown to be associated with a poor prognosis in many cancers. Recently, it has been demonstrated that overexpression of the ErbB2 protein in osteosarcoma is associated with the presence of pulmonary metastasis and decreased survival. To further investigate the role of ErbB2 overexpression in pulmonary metastasis of osteosarcoma, its expression in the primary and metastatic lesions of the same osteosarcoma patients was compared. METHODS: We compared the expression levels of ErbB2 receptor protein between the biopsy samples and pulmonary metastatic lesions in each of 19 patients with osteosarcoma who had not presented with metastasis at diagnosis. All archival materials from patients were retrieved and stained with monoclonal antibody CB11 to detect ErbB2 protein. RESULTS: The rate of overexpression was significantly lower in the pulmonary metastatic tumors than in the biopsy samples (11 versus 42%; p = 0.03). Among 8 patients who had shown increased levels of ErbB2 in the biopsy samples, 7 exhibited complete absence of ErbB2 in the pulmonary metastatic lesions. Overall loss of ErbB2 expression was noted in 14 of 19 patients as the initial tumor became metastatic. CONCLUSIONS: It is suggested that overexpression of ErbB2 decreases within individual osteosarcomas as they become metastatic. Overexpression of ErbB2 may not play an important role in the development of pulmonary metastases of osteosarcoma. Further data are needed before ErbB2 can be used in making clinical decisions for osteosarcoma patients. PMID- 11408806 TI - Establishment and characterization of a new cell line (SKS) from neuroendocrine small cell carcinoma of the uterine cervix and its chemosensitivity. AB - A new cell line (SKS) established from ascites of a patient with neuroendocrine small cell carcinoma of the uterine cervix had a good tumorigenicity and caused marked peritoneal dissemination, and was also highly sensitive to gemcitabine in an in vitro chemosensitivity test. SKS cells were small round cells with a high nuclear/cytoplasmic (N/C) ratio and grew into colony-like aggregates, forming spherical aggregates of floating cells. The population doubling time was 44 h. The number of chromosomes ranged from 50 to 56. On examination of the ultrastructure, membrane-bound dense-core neurosecretory-type granules were observed in the cytoplasm. Neuron-specific enolase (NSE) was immunocytochemically positive in the cytoplasm, and 9.3 ng/ml of NSE was detected in the cell culture supernatant. Human papillomavirus was not detected. In the p53 gene, a 3-bp deletion, AAC (Asn), was detected at codon 131 in exon 5. SKS exhibited good tumorigenicity, and the tumor doubling time was 11 days. Intraperitoneal injection of the cells caused peritoneal dissemination, and marked ascites formation was observed. SKS was highly sensitive to gemcitabine, and the 50% growth inhibitory concentration was 30 nM. SKS cells are useful as a model of neuroendocrine small cell carcinoma of the cervix, and chemotherapy using gemcitabine may possibly be effective in this malignancy. PMID- 11408807 TI - Dityrosine crosslinking of fibrin(ogen) may offer additional protection of tumor cells against immune killing. PMID- 11408810 TI - General considerations on distant metastases from head and neck cancer. AB - Mortality in head and neck cancer is due to locoregional disease, distant metastases or intercurrent disease. As treatment of the primary tumor and cervical metastases has improved, the proportion of deaths from co-morbidity and from distant metastases has increased. Distant metastases almost invariably herald a poor prognosis in head and neck cancer with an average survival of 4.3 7.3 months and treatment is usually palliative. Reliable detection is important to prevent inappropriate treatment. The risk is related to the site, stage and histology of the primary tumor and the presence of cervical metastases. Early detection and treatment of cervical metastases may prevent distant metastases. Accurate staging of tumors helps to identify high-risk tumors that should be specifically investigated for distant metastases. PMID- 11408811 TI - The biology of distant metastases in head and neck cancer. AB - The detection and treatment of metastatic cancer continues to be a challenge for the head and neck oncologist. Unfortunately, head and neck cancer patients who develop distant metastases commonly present late in their course and rapidly succumb to their disease, despite advances in imaging technologies and increased sophistication of biochemical analyses. The development of a rational approach to detection and treatment of metastatic head and neck cancers should begin with an understanding of how these tumors occur and which patients are at greatest risk for developing them. This article presents an overview of the biological processes resulting in the speed of a malignancy from one site to another, with particular attention to head and neck carcinomas. The basic histopathologic, immunology and biochemical abnormalities associated with the development of these secondary tumors are also discussed. PMID- 11408812 TI - Incidence and sites of distant metastases from head and neck cancer. AB - The incidence of distant metastases in head and neck squamous cell carcinoma (SCC) is relatively small in comparison to other malignancies. Distant metastases adversely impact survival and may significantly affect treatment planning. The incidence of distant metastases is influenced by location of the primary tumor, initial T and N stage of the neoplasm, and the presence or absence of regional control above the clavicle. Patients with advanced nodal disease have a high incidence of distant metastases, particularly in the presence of jugular vein invasion or extensive soft tissue disease in the neck. Primary tumors of advanced T stages in the hypopharynx, oropharynx and oral cavity are associated with the highest incidence of distant metastases. Pulmonary metastases are the most frequent in SCC, accounting for 66% of distant metastases. It may be difficult to distinguish pulmonary metastasis from a new primary tumor, particularly if solitary. Other metastatic sites include bone (22%), liver (10%), skin, mediastinum and bone marrow. An important question remains as to how intensely pre- and postoperative screening for distant metastases should be performed. Preoperative chest X-ray is warranted in all cases. If the primary tumor and nodal status place the patient at high risk for pulmonary metastasis, then preoperative computed tomography scan of the chest should be done. Screening for distant metastases at other sites is usually not indicated in SCC of the upper aerodigestive tract. Postoperatively, annual X-rays of the chest are usually sufficient, but in high-risk situations a chest X-ray performed every 3-6 months may be beneficial. Certain histologic types of primary tumor have greater or lesser propensity to metastasize distantly, and have a different natural history. Adenoid cystic carcinoma metastasizes frequently, even in the absence of extensive local or regional disease. Basaloid squamous cell carcinoma and neuroendocrine carcinomas also metastasize widely. Extensive evaluation for distant metastases is justified for these tumors. Knowledge of the natural history of various neoplasms and the factors that contribute to distant metastases as well as good judgement are essential for cost-effective treatment planning and decision-making with regard to pre- and postoperative evaluation for distant metastases in cancer of the head and neck. PMID- 11408813 TI - Screening tests to evaluate distant metastases in head and neck cancer. AB - Investigation for distant metastases is part of the staging process of a primary tumor or recurrent disease before treatment. The lung is the most frequent site followed by bone and liver. Advanced stage and cervical metastases are the most important predictors of metastases. Almost all distant metastases are associated with lung metastases. Computed tomography scan of the chest is the single most effective investigation. The value of routine screening tests is questionable and merits further investigation. PMID- 11408814 TI - Distant metastases from sinonasal cancer. AB - Distant metastases from sinonasal malignancy are generally a rare event except in the terminal stages of the diseases and many patients die from recurrence before the secondaries become clinically manifest. Sinonasal neoplasia covers a diverse range of pathologies, some of which have a greater tendency to spread than others, in particular adenoid cystic carcinoma, malignant melanoma and some of the sarcomas. Notwithstanding this, the frequency with which systemic metastases occurs is such that screening at presentation could not be regarded as cost effective and is consequently only instituted in the presence of specific symptoms. PMID- 11408815 TI - Distant metastases from nasopharyngeal cancer. AB - Undifferentiated carcinoma is the most frequent nasopharyngeal cancer; it has a typical pathognomonic histological pattern, a close relationship to Epstein-Barr virus (EBV), a peculiar natural history and a good prognosis. It has an early tendency to locally spread to the parapharyngeal space. Nodal involvement is highly frequent (70-90%) and bulky regardless of the size of the primary. Literature reports up to 11% distant metastases at presentation and up to 87% at autoptic studies. Pretreatment work-up should include: personal history, clinical and fiberscopic examination, magnetic resonance imaging (MRI) or computed tomography (CT) scan of the base of the skull and neck, histology of the primary and cytology of neck lumps, bone marrow aspiration and biopsy, and EBV serological profile. Clinical and pathological factors predicting possible distant spread are primary tumor and node extension, and treatment failure. Up to now no reliable predictive biological markers have been identified. After treatment, distant metastases are found in about 30% of patients within 5 years and generally have a bad prognosis. Metastatic nodes above the clavicle, in absence of locoregional failure, aggressively treated with chemoradiotherapy, have a disease-free survival longer than 5 years. The following is the suggested posttreatment work-up for early diagnosis of these salvageable patients: clinical and fiberscopic evaluation every 3 months for 2 years and later on every 6 months; skull base and neck MRI or CT scan, and chest CT scan at 6, 12, 18, 24, 36, 48 and 60 months; EBV serological evaluation. PMID- 11408816 TI - Distant metastases from lip and oral cavity cancer. AB - Distant metastases related to lip carcinomas occur very exceptionally (0.5-2%) and can be expected in cases of advanced tumors with advanced regional disease. Distant metastases from oral cavity carcinomas vary over a broad interval (8-17%) and depend also on the stage of disease. The knowledge of the presence of distant metastases is vital for the planning of further treatment. From a clinical point of view, patients with lip and oral cavity can be divided into a group where the risk of distant metastases is low, and into a high-risk one. In low-risk group patients (stages I, II and III) the risk of the incidence of distant metastases is 3%, and the diagnostic approach should consist of an X-ray of the lungs and liver tests. Further examinations are necessary if there are symptoms suggesting the presence of distant metastases or previous examinations are abnormal. The high-risk group (stage IV) and all patients with locoregional relapse have a risk of distant metastases of approximately 10% and the best treatment consists of a positron emission tomography (PET) scan. If a PET scanner is not available it is recommended to run a computed tomography scan of the lungs and liver tests. If any clinical investigation is abnormal further tests are necessary. PMID- 11408817 TI - Distant metastases from oropharyngeal cancer. AB - Distant metastasis is a significant problem in patients with carcinoma of the oropharynx, occurring in approximately 15-20% off all patients over the course of the disease. It is, however, a relatively uncommon first site of failure, as compared to local and regional recurrence. Distant spread occurs most commonly to the lungs, in patients who present with advanced disease, and especially in those with pathologically proven lymph nodes at multiple levels of the neck or in the lower neck. Metastasis to distant sites also occurs more often in patients who recur locally or in the neck. PMID- 11408818 TI - Distant metastases from laryngeal and hypopharyngeal cancer. AB - A retrospective tumor registry analysis of patients with squamous cell carcinoma (SCC) of the larynx and hypopharynx who were treated with curative intent in the Department of Otolaryngology--Head and Neck Surgery at Washington University School of Medicine and Barnes Hospital between January 1971 and December 1991. In 2,550 patients, the mean age, sex and tumor differentiation did not affect the incidence of distant metastases. The overall incidence of distant metastases was 8.5% (217/2,550 patients) with the following distribution: glottis 4.4%, supraglottis 3.6%, subglottis 14%, aryepiglottic fold 16%, pyriform sinus 17% and posterior hypopharynx 17.6%. The overall 5-year disease-specific survival for distant metastases was 6.4%. Distant metastases were related to advanced local disease (T3 + T4), lymph node metastases at presentation (N+), tumor location (hypopharynx) and locoregional tumor recurrence (p < or = 0.028). A meta-analysis of variables which predispose to a higher incidence of distant metastases indicate that tumor location (hypopharynx > larynx), advanced primary disease (T3 + T4), regional disease (N+), locoregional recurrences, and advanced regional metastases (N2 + N3) are statistically significant. PMID- 11408819 TI - Distant metastases from cervical esophagus cancer. AB - Cancer of the cervical esophagus has a poor prognosis in relation to stage. Correct staging is thus essential in order to establish the prognosis and the treatment program. Distant metastases can involve the lymph nodes (mediastinal and celiac lymph nodes) or they can be extranodal visceral types. Correct lymph node staging can be performed with esophageal endoscopic ultrasonography, computed tomography (CT) scan and, currently, with positron emission tomography (PET) and minimally invasive surgery. For hematogenous metastases, CT scan and PET are mainly used, as well as minimally invasive surgery, with the eventual aid of intraoperative ultrasonography. PMID- 11408820 TI - Distant metastases from salivary glands cancer. AB - Patients who present with malignant salivary glands should at their initial assessment have an X-ray of the chest to exclude the possibility of distant metastases. Patients who have other symptoms, bone pain etc., should be appropriately investigated. The likelihood of patients developing distant metastases is associated with high-grade tumors, most commonly adenoid cystic carcinoma, high-grade mucoepidermoid carcinoma, salivary duct carcinoma and tumors sited in the submandibular gland, posterior tongue and pharyngeal tumors. Patients who have had a high-grade tumor treated and survived without locoregional recurrence have the same risk of developing distant metastases as those patients who have locoregional recurrence. Other histological types of salivary tumors are associated with a lower risk of developing distant metastases but a real risk remains lifelong. It is recommended that all patients who have a malignant salivary gland tumor treated, any histology, should be followed up and clinically assessed at least once every 12 months for life. PMID- 11408821 TI - Distant metastases from thyroid and parathyroid cancer. AB - Thyroid cancer represents a unique biological tumor where even with the high incidence of distant metastases, the overall prognosis is not as poor as many other human cancers. The overall long-term survival in patients presenting initially with distant metastasis is approximately 50%. The overall incidence of distant metastases varies between 10 and 35%, depending upon the histology. The overall incidence is directly related to various histologies - being least in papillary thyroid carcinoma (10%) and highest in Hurthle cell tumor (33%). The incidence of distant metastases is also very high in patients with medullary and anaplastic thyroid cancer. The incidence of distant metastases at the time of initial presentation in differentiated thyroid cancer is approximately 4%. In high-risk patients - especially in patients with extrathyroidal extension or massive nodal metastasis - the distant metastases can be evaluated after total thyroidectomy with radioactive iodine ablation. Pulmonary metastases are very common in young individuals, but they are extremely well treated and the mortality from distant metastases in this group is very low. However, distant metastases in patients with poorly-differentiated carcinoma have a poor prognosis. In high-risk patients, generally a total thyroidectomy should be undertaken so that the patient can undergo radioactive iodine dosimetry and ablation as indicated. The surveillance in patients with thyroid cancer includes: close clinical follow-up, chest X-ray, and radioactive iodine dosimetry. Thyroglobulin is commonly used as a prognostic marker in patients having undergone total thyroidectomy. The incidence of distant metastases in medullary thyroid cancer is high, mainly to the lung and liver. Persistent hypercalcitonemia is an indication of regional or distant metastases. A variety of diagnostic tests are helpful, such as octreotide scanning, computed tomography scan, magnetic resonance imaging and positron emission tomography scan. Laparoscopy to evaluate the surface of the liver is also an important investigation to detect distant metastases. The incidence of distant metastases is very high in patients with anaplastic thyroid cancer, but most of the time the outcome depends on the locoregional recurrence and massive disease in the central compartment. The parathyroid cancer is quite rare, less than 1%, in patients undergoing parathyroidectomy. The diagnosis of parathyroid cancer is made by pathological features but the most certain method of diagnosis of a malignant tumor of the parathyroid is the identification of secondary deposits. The incidence of distant metastasis is difficult to determine due to the rarity of this condition, but the most common site is the lung. Patients with distant metastasis have recurrent progressive hypercalcemia along with high parathormone level. PMID- 11408822 TI - Distant metastases from ear and temporal bone cancer. AB - Cancers of the temporal bone are rare. Cervical metastases occur in approximately 10% of cases and are much more likely once disease extends beyond the confines of the temporal bone. Nonlymphatic spread of squamous cell carcinoma is usually a late event resulting in metastatic deposits in the lung, bone, liver and brain. This chapter discusses detection of distant metastases and provides a recommended schedule for interval patient evaluation. PMID- 11408823 TI - Noncervical lymph node metastasis from head and neck cancer. AB - Nonregional lymph node dissemination must be classified as distant metastasis but axillary and mediastinal metastases can be part of a regional dissemination of the disease. Metastases to lymph nodes of the upper mediastinum are very common among patients with subglottic, hypopharynx and thyroid carcinomas. Axillary metastases are found at autopsy in 2-9% of the patients who died of head and neck squamous cell carcinoma (SCC) and are frequently associated with skin implantation in aggressive recurrent head and neck carcinomas. The possible explanations for this location of metastasis were retrograde dissemination due to lymph system blockage, further tumor dissemination after a parastomal recurrence, hematogenous dissemination, and metastasis from a second primary tumor. Patients with distant metastasis have been considered incurable and only palliative treatment was instituted. Treatment planning for cases with axillary metastasis must take in consideration the likelihood of other regional recurrences and/or distant metastasis. Also, the presence of a second primary tumor must be ruled out. Whenever axilla is the only site of cancer recurrence, a standard axillary dissection must be considered. Upper mediastinal metastases from subglottic and hypopharyngeal cancer are managed by paratracheal and mediastinal dissection through the neck and postoperative radiotherapy. PMID- 11408824 TI - Proposal of standardization on screening tests for detection of distant metastases from head and neck cancer. AB - A standardized approach to screening for distant metastases must be flexible. This reflects the fact that the issues involved have not been adequately studied to allow evidence-based recommendations. Effective and responsible application of screening for distant metastasis would improve our ability to council patients regarding important therapeutic decisions. It would also have important consequences on healthcare economics. Screening recommendations should reflect the stage of the primary tumor and the histologic cell type because these two parameters most clearly correlate with the risk for distant metastases. The most commonly encountered distant metastases are pulmonary. The most sensitive validated screening technique is just computed tomography (CT). Advanced technologies such as simultaneous positron emission tomography/CT may replace these prior technologies in the near future. Recommendations for routine screening following curative treatment are subjective at this time. Careful history and physical examination remains the basis for the follow-up evaluation. The sensitivity and cost effectiveness of imaging studies in this setting remains unstudied and contentious. PMID- 11408825 TI - The treatment of distant metastases in head and neck cancer--present and future. AB - At the present time the occurrence of distant metastases in patients with head and neck squamous cell carcinoma means that lifespan is measured in months. In most instances treatment is purely palliative. Isolated lung metastasis can be successfully removed with long-term disease control in selected patients. Radiotherapy can be useful for palliation of bone metastases and occasionally lung or brain metastases. Chemotherapy does not have a major impact at the present time except for the treatment of metastases from nasopharyngeal cancer. Palliative symptomatic care, along with appropriate pain control, is essential since pain management is very important in these patients. A significant change in the survival of patients with head and neck cancer is only likely to occur by the development of new approaches to treatment. Blocking tumor angiogenesis and treatment based on genetic abnormalities or cell surface receptors offer the two strategies that are most likely to be successful. PMID- 11408826 TI - Effects of taurine on cerulein-induced acute pancreatitis in the rat. AB - Taurine, or 2-aminoethane sulfonic acid, is an intracellular amino acid and has been suggested to have a function in protecting biological systems from oxidative tissue damage. The aim of this study was to determine the effect of taurine against cerulein-induced acute pancreatitis in rats. Acute pancreatitis was induced by administering three subcutaneous injections of cerulein (40 microg/kg body weight) at 1-hour intervals, while taurine was administered intravenously at graded doses (30, 100, or 300 mg/kg, respectively) following the first cerulein injection. The severities of pancreatitis and lung injury were determined by measuring biochemical parameters, tissue myeloperoxidase (MPO), and histological changes. To clarify the mechanism of taurine, serum IL-1beta and TNF-alpha levels and tissue concentrations of malondialdehyde (MDA) were evaluated. In cerulein induced acute edematous pancreatitis, treatment with taurine significantly decreased hyperamylasemia, tissue MPO, pancreatic edema, and the extent of pancreatic and pulmonary injury. Taurine decreased MDA concentration in the pancreas and lung, but not the serum cytokine concentration. We would conclude that taurine has beneficial effects in cerulein-induced acute pancreatitis and lung injuries by preventing the production of oxygen free radicals. PMID- 11408827 TI - Pharmacological characterization of a novel 5-HT4 receptor agonist, TS-951, in vitro. AB - The pharmacological effect of a novel selective 5-HT4 receptor agonist, TS-951 (N [endo-8-(3-hydroxypropyl)-8-azabicyclo[3.2.1]oct-3-yl]-1-isopropyl-2-oxo-1,2 dihydro-3-quinolinecarboxamide) was investigated in vitro. TS-951 potently inhibited specific [3H]GR113808 binding both in guinea-pig striatum and in mouse brain. The affinity of TS-951 for the 5-HT4 receptor was higher than those of other agonists, 5-HT, cisapride, mosapride and renzapride. On the longitudinal muscle of the guinea-pig ileum, TS-951 caused a concentration-dependent increase in the amplitude of electrically induced submaximal twitch contractions. On the longitudinal muscle of the guinea-pig distal colon, TS-951 also caused concentration-dependent contractions. TS-951 is a high-affinity, selective and potent 5-HT4 receptor agonist. This compound therefore can be considered as a useful pharmacological tool for investigating 5-HT4 receptor-mediated events. PMID- 11408828 TI - Tas-301, a new synthetic inhibitor of neointimal thickening after balloon injury, inhibits calcium-dependent signal transduction and cytoskeletal reorganization. AB - We previously demonstrated that a recently synthesized drug, TAS-301 [3-bis(4 methoxyphenyl)methylene-2-indolinone], inhibited neointimal thickening after single-balloon injury to the rat common carotid artery by inhibiting both the migration and proliferation processes of vascular smooth muscle cells (VSMCs). The purpose of this current study was to elucidate the possible mechanism of action for its inhibition of the migration process of VSMCs. We also determined the efficacy of TAS-301 on second neointimal formation 14 days after a double balloon injury to the rat common carotid artery. Neointimal thickening, 14 days after second balloon injury, was reduced by the oral administration of TAS-301 in a dose-dependent manner. In in vitro assays using rat VSMCs, Western blot analysis showed that TAS-301 inhibited platelet-derived growth factor (PDGF) induced tyrosine phosphorylation of both focal adhesion kinase and paxillin. Tyrosine phosphorylation of these proteins depended on the increment of intracellular calcium concentration ([Ca2+]i). The PDGF-induced elevation of [Ca2+]i and activation of Ca2+/calmodulin-dependent protein kinase II (CaM kinase II) were also inhibited by TAS-301. Additionally, TAS-301 at 10 micromol/l reduced the extent of F-actin stress fiber depolymerization observed in response to PDGF. These results indicate that TAS-301 reduced the intimal thickening after denudation to a pre-existing lesion to the same extent as it reduced that after a single-balloon injury to the normal artery. Furthermore, the results of our in vitro experiments suggest that antimigratory mechanisms of TAS-301 that contribute to preventing the intimal thickening might be mediated by drug inhibition of Ca2+ -dependent signal molecules and the following cytoskeletal depolymerization. PMID- 11408829 TI - Effect of cyclooxygenase-2 inhibitor (celecoxib) on the infarcted heart in situ. AB - Several attempts have been made to replace aspirin with compounds without gastric toxicity; a cyclooxygenase-2 (COX-2) inhibitor, celecoxib, and a nitric oxide aspirin, NCX-4016, have been developed for this purpose. This paper compares effects of celecoxib, NCX-4016 and aspirin on production of prostacyclin (PGI2) and thromboxane A2 (TXA2) and activation of the inducible form of nitric oxide synthase (iNOS) in infarcted heart in situ. Aspirin was most effective in reducing myocardial PGI2 synthesis and formation of TXA2. Myocardial effects of celecoxib resemble those of NCX-4016, although the two compounds have different modes of action. PMID- 11408830 TI - Honokiol and magnolol selectively interact with GABAA receptor subtypes in vitro. AB - Honokiol and magnolol have been identified as modulators of the GABAA receptors in vitro. Our previous study suggested a possible selectivity of honokiol and magnolol on GABAA receptor subtypes. This possibility was examined in the current study by 3H-muscimol and 3H-flunitrazepam binding assays on various rat brain membrane preparations and human recombinant GABA(A) receptor subunit combinations expressed by the Sf-9/baculovirus system. Generally, honokiol and magnolol have a similar enhancing effect on (3)H-muscimol binding to various membrane preparations in nonsaturation binding assays. Honokiol and magnolol preferentially increased (3)H-muscimol binding to hippocampus compared to cortex and cerebellum (with a maximum enhancement of 400% of control). As for subunit combinations, honokiol and magnolol have a more potent enhancing effect on alpha2 subunit containing combinations (with a maximum enhancement of 400-450% of control). This action was independent of the gamma subunit. In saturation binding assays, magnolol affected either the number of binding sites (ca. 4-fold on alpha2 containing combinations) or the binding affinity (on alpha1 containing combinations) of (3)H-muscimol binding to various GABAA receptor subunit combinations. In contrast, honokiol increased only binding sites on alpha2beta3gamma2s and alpha2beta3 combinations, but both the number of binding sites and the binding affinity on alpha1beta2gamma2S and alpha(1)beta2 combinations. These results indicate that honokiol and magnolol have some selectivity on different GABAA receptor subtypes. The property of interacting with GABAA receptors and their selectivity could be responsible for the reported in vivo effects of these two compounds. PMID- 11408831 TI - Diazepam stimulates migration and phagocytosis of human neutrophils: possible contribution of peripheral-type benzodiazepine receptors and intracellular calcium. AB - In isolated human neutrophils, diazepam (10 nmol/l to 10 micromol/l) concentration-dependently increased migration and phagocytosis. Diazepam-induced migration and phagocytosis were inhibited by the peripheral benzodiazepine receptor (PBR) antagonist PK11195 (10 micromol/l). The PBR agonist Ro5-4864 (10 nmol/l to 10 micromol/l) did not affect migration but slightly enhanced phagocytosis, while clonazepam, which binds to the central-type benzodiazepine receptors but has no affinity for PBRs, was ineffective on both parameters up to 10 micromol/l. Phagocytosis induced by diazepam or Ro5-4864 was inhibited by the Ca2+ channel blocker L-verapamil (10 micromol/l), which however did not affect the action of diazepam on migration. Competition binding experiments performed by fluorescent staining of PBRs showed that diazepam directly interacts with PBRs on human neutrophils. Both diazepam and Ro5-4864 (10 nmol/l to 10 micromol/l) induced a rise of intracellular free Ca2+ concentrations ([Ca2+]i), which was inhibited by PK11195 (10 micromol/l) and L-verapamil (10 micromol/l) and prevented by extracellular Ca2+ chelation with EGTA (5 mmol/l). In conclusion, experimental evidence indicates that in human neutrophils diazepam stimulates both migration and phagocytosis through activation of PBRs. Diazepam-induced [Ca2+]i changes depend on a PBR-operated, L-verapamil-sensitive increase in the plasma membrane permeability and subsequent extracellular Ca2+ entry, and contribute to diazepam-induced phagocytosis. On the contrary, the effect of diazepam on migration seems to occur through Ca2+ -independent mechanisms. PMID- 11408832 TI - The pharmacological effect of omeprazole on water gastric emptying: A study based on an impedance measure. AB - The present study was performed to validate the accuracy of a self-designed applied-potential tomograph (APT) in measuring the cross area of a studied object and to assess the effect of omeprazole premedication on water gastric emptying (GE), based on APT. Twelve electrodes were evenly placed in a circular array around the studied subjects. Ten electrodes in a rotated order recorded the electrical current injected into paired electrodes. Based on tomography, averaged signals of changed resistivity were constructed to display the area of interest and GE curve. Six beakers of various diameters were respectively placed into a cylindrical perspex tank which was filled with saline to measure their cross areas via computer-generated diagrams of APT. One hour after either omeprazole (20 mg) or placebo premedication, 15 healthy males were ordered to consume 500 ml of test water to assess emptying for 40 min. Within 3 days, a similar procedure was repeated using the counterpart premedication. The true cross areas of the 6 beakers are 2.01, 15.9, 18.8, 30.19, 38.48 and 63.61 cm(2), whereas those obtained by APT were 7.9 +/- 2.9, 16.7 +/- 3.3, 22.4 +/- 4.9, 28 +/- 4.8, 48.7 +/ 7.6 and 67 +/- 6.1 cm(2), respectively (r = 0.98, p < 0.001). Valid emptying data were obtained in 73.3 and 86.6% of subjects, following placebo and omeprazole, respectively (not significant). The half emptying times were 12.7 +/- 5.1 min for the placebo-treated group and 10.5 +/- 3.6 min for the omeprazole treated group, respectively (p < 0.05). The areas under the emptying curve were 1,611.5 +/- 357.6 and 1,317.3 +/- 316.7 arbitrary units, respectively (p < 0.01). In conclusion, our APT system is accurate for large-area measurements; acid inhibition before APT measurement does not increase the success rate but enhances water emptying. The interpretation of impedance-obtained GE should consider the acid-inhibitory effect. PMID- 11408833 TI - Effects of peroxynitrite on the reactivity of diabetic rat aorta. AB - Endogenous nitric oxide (NO) reacts with superoxide to form peroxynitrite, which is capable of either oxidizing or nitrating various biological substrates. We compared the vasodilatory effect of exogenous peroxynitrite with the effects of decomposed peroxynitrite or sodium nitrite in precontracted aorta isolated from streptozotocin-induced diabetic and age-matched control rats. Peroxynitrite (10 nmol/l to 300 micromol/l) produced a concentration-dependent relaxation in aortic rings with or without endothelium. Relaxation was also observed with a higher concentration of its decomposition product or sodium nitrite, although these relaxations were considerably slower and with reduced sensitivity. Endothelium containing rings were less sensitive to the vasorelaxant effect of peroxynitrite than the endothelium-denuded rings in control (pD(2) was 5.19 +/- 0.06 in rings with endothelium and 5.86 +/- 0.03 in rings without endothelium, p < 0.01) but not in diabetic aorta (pD(2) was 5.97 +/- 0.05 in rings with endothelium and 6.12 +/- 0.06 in rings without endothelium, p > 0.05). The maximum relaxation to peroxynitrite also increased in diabetics, but did not change by removal of the endothelium either in diabetic or control rings. Diabetes did not alter the relaxations elicited by both decomposed peroxynitrite and sodium nitrite. Peroxynitrite-induced relaxation was not inhibited by diethylenetriaminepentaacetic acid, an inhibitor of hydroxyl radical formation. Pretreatment with peroxynitrite (1 micromol/l, 15 min) significantly suppressed the phenylephrine-induced tone and acetylcholine-stimulated endothelium-dependent relaxation, both effects were more pronounced in diabetic than in control aorta. The increased responsiveness of diabetic vessels to exogenous peroxynitrite seems to be related to depressed basal NO bioavailability and may be considered as a compensatory way against activated contractile mechanisms of diabetic vascular smooth muscle. PMID- 11408834 TI - Assessment of psychological distress in the setting of medical disease. AB - The biopsychosocial model of disease has recently been depicted as the basis for a renewed emphasis on the multiaxial diagnostic system of the DSM-IV. The authors challenge this stance, underscoring the clinical inadequacies of the DSM-IV in the setting of medical disease, particularly the chapters concerned with somatoform disorders and psychological factors affecting medical conditions. Diagnostic criteria which are based on the clinical insights derived from psychosomatic research in the past decades may offer new opportunities to psychosomatic medicine and consultation-liaison psychiatry. The development of the Diagnostic Criteria for Psychosomatic Research (DCPR), encompassing alexithymia, type A behavior, irritable mood, demoralization, disease phobia, thanatophobia, health anxiety, illness denial, functional somatic symptoms secondary to a psychiatric disorder, persistent somatization, conversion symptoms and anniversary reaction, is described. Preliminary results obtained with the combination of DSM and DCPR criteria appear to be promising. PMID- 11408835 TI - Psychological evaluation after cardiac transplantation: the integration of different criteria. AB - BACKGROUND: The psychological evaluation of patients undergoing cardiac transplantation is currently based on DSM-IV criteria. An alternative diagnostic and conceptual framework has been proposed by an international group of psychosomatic investigators. The aim of this study was to compare these new criteria (Diagnostic Criteria for Psychosomatic Research, DCPR) with DSM-IV in a population where a high prevalence of psychological problems is expected (heart transplanted patients). METHOD: 129 consecutive patients who underwent heart transplant surgery were assessed according to DSM-IV and DCPR criteria. RESULTS: The results showed a higher number of diagnoses made using the DCPR than with the use of the DSM-IV. At least one DCPR diagnosis was found in 85 (66%) patients, whereas at least one DSM diagnosis was present in 23 (18%) patients. The number of DCPR diagnoses was almost the triple of DSM criteria. While patients who were given a DSM diagnosis frequently had additional DCPR diagnoses, many patients with DCPR criteria did not fulfill any DSM criteria. Four DCPR syndromes appeared to be particularly frequent: demoralization, type A behavior, irritable mood and alexithymia. CONCLUSIONS: The joint use of DSM and DCPR criteria was found to improve the identification of psychological factors which could result in a worsening of quality of life in heart-transplanted patients. PMID- 11408836 TI - Criterion-related validity of the diagnostic criteria for psychosomatic research for alexithymia in patients with functional gastrointestinal disorders. AB - BACKGROUND: Because of criticism made against psychiatric assessment of somatization, alternative Diagnostic Criteria for Psychosomatic Research (DCPR) have been proposed by an international group of psychosomatic investigators. One of these criteria concerns the alexithymia construct. The main aim of the present study was to investigate the criterion-related validity of the DCPR for alexithymia syndrome (DCPR-A). A secondary aim was to explore the relationship between alexithymia and depressed mood. METHOD: The study included 190 consecutive outpatients with functional gastrointestinal disorders. Alexithymia was assessed by means of the DCPR-A and the 20-item Toronto Alexithymia Scale (TAS-20). Depressed mood was assessed on the basis of the Depression subscale of the Hospital Anxiety and Depression Scale (HDS) and the DSM-IV criteria. RESULTS: The sensitivity of the DCPR-A together with the TAS-20 was 70.2%, specificity was 81.6%, positive predictive power 88.9%, negative predictive power 66.0% and overall hit rate 46.8%. DCPR-A positives scored significantly higher than DCPR-A negatives on the TAS-20 scores (t = 9.86, p < 0.001). The TAS-20 was not associated with any measure of depression, while the DCPR-A was significantly associated with the HDS and DSM-IV criteria. CONCLUSIONS: The DCPR-A had adequate sensitivity, specificity and accuracy, thus proving that the criterion has good validity. The differences in association found between alexithymia and depression suggest that they may be due to differences in the methods of investigation rather than to the constructs per se. PMID- 11408837 TI - The growing commercial value of the male body: a longitudinal survey of advertising in women's magazines. AB - BACKGROUND: With the advances of feminism, men have gradually relinquished their once-exclusive 'masculine' roles as fighters and breadwinners. In response to this change, the male body may have gained in relative importance as one of the few surviving marks of masculinity. We hypothesized that these trends might be quantified by using a commercial measure such as advertising. METHODS: We examined the proportion of exposed male and female bodies portrayed in advertisements between 1958 and 1998 in two leading American women's magazines. RESULTS: In both magazines, the proportion of undressed women in the advertisements has changed little over the last 40 years, whereas the proportion of undressed men has increased dramatically, especially since the early 1980s. CONCLUSIONS: Trends in commercial advertising offer tentative support for the hypothesis that the male body is increasing in importance as a mark of masculinity--at least as judged from the actions of advertisers seeking to influence women's attitudes. PMID- 11408838 TI - Food exposure in patients with bulimia nervosa. AB - BACKGROUND: The aim of the study was to investigate psychological and physiological responses of bulimic patients to the repeated presentation of food cues. METHOD: On 2 subsequent days, 48 bulimic women (DSM-IV) were confronted with high- or low-caloric food for 20 min. A control group (n = 24) was exposed to high-caloric food once. Blood sugar levels were manipulated with a glucose load. RESULTS: High-caloric food only elicited increases in urge to binge, subjective and physiological stress in the first session. During the second session, reported urge to binge and subjective stress was significantly lower. Bulimic patients confronted with high-caloric food showed higher subjective and physiological stress in the first session compared to bulimic patients confronted with low-caloric food. In the second session, they reported more subjective stress and urge to binge, compared to the low-caloric group. Blood sugar levels did not affect psychological and physiological responses. DISCUSSION: The results are discussed in terms of the conditioning model of binge eating model, habituation models and implications for exposure therapy. PMID- 11408839 TI - Outcome predictors in anorectic patients after 6 months of multimodal treatment. AB - BACKGROUND: Anorexia nervosa is a severe disorder that often responds poorly to treatment. At this time, little is known about pretreatment predictors of response. METHODS: A sample of 42 restrictor type anorectics was tested at the assessment phase and after 180 days retested using the Temperament and Character Inventory and Eating Disorder Inventory 2 along with other clinical evaluation instruments. After 180 days of treatment with multimodal 'network' therapy, the patients were divided into two groups. The first group included patients who showed relevant clinical improvement; the second group included patients considered 'not yet responding'. Data collected from the not-yet-responding group were compared by the t test with the other group's data to evaluate prognostic indexes. RESULTS: Diagnosis of personality disorder, which afflicted about 50% of patients, seemed not to be a relevant prognostic factor. However, a lower novelty seeking was characteristic of the nonresponder group. Higher levels of asceticism and maturity fears also characterized the nonresponder group. CONCLUSION: Present data suggest some elements that could be useful to focus pharmacotherapy, psychotherapy and family counseling on the current psychopathology of each patient. PMID- 11408840 TI - Changes in conflict, symptoms, and well-being during psychodynamic and cognitive behavioral alcohol inpatient treatment. AB - BACKGROUND: According to Grawe's psychological therapy approach, conflict reduction can be expected not only in psychodynamic, but also in cognitive behavioral therapy (CBT). This was tested in an effectiveness study. Changes in cognitive conflicts, along with those of symptom severity and well-being were analyzed during alcohol inpatient treatment. METHODS: Four times during treatment, groups of patients receiving psychodynamic therapy (n = 45 patients) or CBT (n = 49 patients) were measured and compared. Lauterbach's Online Conflict Test was used to measure conflict. Symptom severity and well-being were measured using questionnaires. RESULTS: Results showed significant conflict decrease in both groups with a tendency towards faster reduction under CBT. There was also significant change in symptom severity and well-being in both groups with no difference regarding reduction gradient. Moreover, patients in the psychodynamic treatment group exhibited lower symptom scores at treatment begin which may be a consequence of clinical group assignment. CONCLUSIONS: In general, the findings confirmed Grawe's theoretical assumptions. Generalizability to other diagnostic groups and other clinical settings remains to be tested. PMID- 11408841 TI - Association of serum nitric oxide levels with depressive symptoms: a study with end-stage renal failure patients. AB - BACKGROUND: Nitric oxide (NO) is a soluble gas produced by the activity of an enzyme found in neurons. It has been implicated in a great number of normal physiological functions (such as noradrenaline and dopamine release, memory and learning, regulation of the cerebrovascular system, modulation of wakefulness, modulation of nociception, olfaction, food intake and drinking) as well as pathologies (Alzheimer's, Huntington's disease, cerebral ischemia, stroke). Two reports have addressed the involvement of NO in depression. METHODS: The objective of the study was to examine the association between NO and specific depressive symptoms. For this purpose, in a sample of 28 end-stage renal failure patients (who have increased NO levels), we tested the hypothesis that the subgroup of patients with these specific depressive symptoms was differentiated from the patients without these symptoms with regard to serum levels of NO metabolites. The depressive symptoms were assessed using the Zung self-rating scale. RESULTS: Our study revealed an association of NO with the following depressive symptoms: sexual dysfunction, weight loss, psychomotor retardation, indecisiveness and irritability. CONCLUSION: The association between NO system and symptoms of depression does not necessarily imply a pathogenetic association between NO and depressive disorder. Further research is needed to verify these findings and study their possible pathogenetic implications. PMID- 11408842 TI - Sexual dysfunction in male psychiatric outpatients: validity of the Massachusetts General Hospital Sexual Functioning Questionnaire. AB - BACKGROUND: The purpose of this study was to determine the frequency of sexual dysfunction in male outpatients with concomitant psychiatric and medical conditions and to establish the validity of the Massachusetts General Hospital Sexual Function Questionnaire (MGH). METHODS: Male VA outpatients were given two sexual function questionnaires, a 36-item Changes in Sexual Functioning Questionnaire (CSFQ) and the 5-item MGH. Medical records were evaluated for diagnosis, medications and illnesses that might affect sexual functioning. RESULTS: Among 100 outpatient respondents, the CSFQ showed that 90% and the MGH showed that 77% experienced sexual dysfunction. Forty percent of patients reported total absence of sexual function. Good concurrent validity was established between the MGH and CSFQ. CONCLUSIONS: Sexual dysfunction is highly prevalent in psychiatric outpatients. The MGH is a valid clinical measure to determine sexual dysfunction. PMID- 11408843 TI - New developments in the treatment of psoriasis. AB - During the previous 10 years, important innovations have been developed in the treatment of psoriasis. Calcipotriol has been introduced in most countries already 10 years ago and more recently 2 other vitamin D(3) analogues have become available: calcitriol and tacalitol. In some countries, the topical retinoid tazarotene has been registered as a routine treatment in psoriasis. Evidence based data on the efficacy and safety of combination treatments are accumulating. In particular, the combination of calcipotriol and topical corticosteroids has been shown to be very effective and safe. The combination of systemic treatments is controversial. In individual patients, the risk of combining major therapies should be balanced against the need to enhance efficacy in individuals. New leads in experimental treatments are new vitamin D(3) analogues and new retinoids as well as 4-hydroxylase- and 24-hydroxylase inhibitors. Important innovations in immunomodulatory treatments are: tacrolimus, ascomycine, anti-CD4, anti-CD25, peptide T and LFA3TIP. PMID- 11408844 TI - Studies on the alpha-crystallin target protein binding sites: sequential binding with two target proteins. AB - PURPOSE: alpha-Crystallin belongs to a class of small heat shock proteins and is shown to prevent aggregation of several proteins. We have shown that the temperature-induced structural perturbation leads to several fold enhanced activity. The purpose of this study was to investigate the availability and specificity of the hydrophobic sites that might become available at elevated temperatures. Specifically, we address the following question: Is there an increased exposure of fixed number of hydrophobic sites as a function of temperature or does a new set of sites become available at elevated temperatures? METHODS: alpha-Crystallin target protein complexes were made at two different temperatures and this complex was investigated for its chaperone-like activity towards the same target protein and also other target proteins. DTT-induced aggregation of insulin, alpha-lactalbumin, thermal aggregation of betaL- and gamma-crystallin, and photo-aggregation of gamma-crystallin were used as model systems. Increased light scattering was used to monitor the progress of aggregation. RESULTS: alpha-Crystallin target protein complex prepared at 37 degrees C temperature was effective against thermal aggregation of betaL crystallin as well as non-thermal aggregation at elevated temperatures. However, the complex prepared at high temperature was ineffective at lower temperatures as well as with other target proteins at both temperatures. CONCLUSIONS: More target protein binding sites become available at elevated temperatures. The sites available at low temperature are a subset of the total sites available at elevated temperatures. PMID- 11408845 TI - The clinical pharmacology of the oral fluoropyrimidines. PMID- 11408846 TI - Obstetrics and gynecology at the crossroads--again? Still! AB - It seems, if one can believe presidential addresses, as if our specialty is always at some crossroads or other. In this "opinion piece," uniquely, I don't complain about managed care. I do attempt to identify some of the issues that will be of import for obstetrics and gynecology in the near and not-so-near futures. With regard to research, we await breakthroughs, for example, in the early detection of ovarian cancer, so as to finally be able to improve outcomes. A problem, though, is our failure to focus enough effort on developing a cadre of clinician scientists, who can work out research findings with direct clinical application; this is an issue with which the specialty needs to come to grips. Regarding education, I believe we need to refocus from what type of practitioners we might want to produce to best meet the needs of our patients. The bottom line should be more flexibility in training and emphasis on clinical competence, so that excellent practitioners with competence across the breadth of our specialty are available to provide a full range of appropriate women's health care. The concept of "women's health" is controversial and evolving rapidly. Review of several available sources suggests that reproductive medicine will remain an important component of women's health but that our specialty must now evolve to include other areas, as the major health problems of women change. We need to shift from an organ-based paradigm to a more holistic view, reflecting the woman centered focus for our specialty. This "paradigm shift" will need to entail continuation of first-rate surgical and obstetrics services but will be expanded to include a full range of services, probably offered by highly trained and competent individuals with specific areas of expertise, practicing in a multi single-specialty group. Obstetrics and gynecology at the crossroads, indeed! PMID- 11408847 TI - The effects of intrapartum magnesium sulfate therapy on fetal serum interleukin 1beta, interleukin-6, and tumor necrosis factor-alpha at delivery: a randomized, placebo-controlled trial. AB - OBJECTIVE: Elevated levels of inflammatory cytokines in the fetus have been linked to neurologic morbidities in preterm neonates. Magnesium sulfate is currently being studied in clinical trials as a potential fetal neuroprotective agent. The purpose of this study was to determine whether intrapartum magnesium sulfate therapy has an effect on the umbilical venous concentrations of interleukin-1beta, interleukin-6, and tumor necrosis factor-alpha at delivery. STUDY DESIGN: Women with singleton gestations >32 weeks with no clinical indications for magnesium sulfate therapy (preeclampsia or tocolysis) and either clinical chorioamnionitis or prolonged rupture of membranes were recruited for the study. Consenting patients were randomly assigned, in a double-blinded fashion, to receive either magnesium sulfate (6-g load then 2 g/hr) or matched volumes of lactated Ringer's solution until delivery. Fetal blood specimens were obtained by aspiration of the umbilical vein after cord clamping but before placental separation. Umbilical cytokine levels were measured with a sensitive and specific immunoassay. RESULTS: Twenty-two patients were randomly assigned to groups and received either magnesium sulfate (n = 11) or placebo (n = 11). There were no differences in the demographic or clinical characteristics between groups. The umbilical venous ionized magnesium concentration was significantly higher in the magnesium sulfate group (2.32 +/- 0.27 mg/dL vs 1.23 +/- 0.15 mg/dL; P <.001). There were no statistically significant differences between groups with respect to umbilical levels of interleukin-1beta (1.5 pg/mL [1.5-58] vs 1.5 pg/mL [1.5-10]; P =.5); interleukin-6 (8.5 pg/mL [1-1000] vs 11.2 pg/mL [1 113]; P =.9); or tumor necrosis factor-alpha (16 pg/mL [7.6-20.3] vs 16.6 pg/mL [8.3-22.2]; P =.5). CONCLUSION: In this pilot study the intrapartum administration of magnesium sulfate does not appear to affect the concentration of inflammatory cytokines in fetal blood at delivery. PMID- 11408848 TI - Does cervical cerclage prevent preterm delivery in patients with a short cervix? AB - OBJECTIVE: Our purpose was to determine whether cerclage placement in women with a short cervix on transvaginal ultrasonography reduces the rate of preterm delivery. STUDY DESIGN: A retrospective cohort study identified patients with an ultrasonographic short cervix (cervical length < or =15 mm) between 14 and 24 weeks' gestation. Cerclage placement was performed at the discretion of the attending physician. Clinical characteristics and outcome with and without cerclage were compared. RESULTS: Seventy patients met inclusion criteria; 25 (36%) underwent cerclage placement. Patients managed with cerclage had a lower gestational age at diagnosis (19.6 weeks vs 21.3 weeks, P <.01) but had a similar median cervical length, presence of funneling, and a history of cervical surgery, in comparison with those managed without cerclage. The rate of spontaneous preterm delivery was not different between groups. Patients with cerclage had a higher rate of preterm premature rupture of membranes than those without cerclage (65.2% vs 36.4%, P <.05). CONCLUSION: Cervical cerclage in patients with a short cervix did not reduce the rate of spontaneous preterm delivery and increased the risk of preterm premature rupture of membranes. PMID- 11408849 TI - A prospective, randomized trial comparing the efficacy of dexamethasone and betamethasone for the treatment of antepartum HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome. AB - OBJECTIVE: This study was undertaken to determine whether dexamethasone or betamethasone is superior for the antepartum treatment of HELLP (hemolysis, elevated liver enzymes, and low platelet count) syndrome. STUDY DESIGN: This prospective, randomized, clinical investigation compared intravenously administered dexamethasone and intramuscularly administered betamethasone in the treatment of gravid women with HELLP syndrome. Efficacy end points included laboratory values (platelet count, lactate dehydrogenase activity, aspartate aminotransferase activity) and clinical parameters (mean arterial pressure, urinary output). RESULTS: Forty patients were enrolled in the study, 19 in the dexamethasone arm and 21 in the betamethasone arm. The adjusted time-averaged changes from baseline were significant for aspartate aminotransferase activity (dexamethasone, -20.4 +/- 9.6 U/L; betamethasone, 9.9 +/- 8.9 U/L; P =.029), mean arterial pressure (dexamethasone, -15.6 +/- 1.4 mm Hg; betamethasone, -8.1 +/- 1.4 mm Hg; P <.001), and urinary output (dexamethasone, 12.9 +/- 8.6 mL/h; betamethasone, -11.9 +/- 8.2 mL/h; P =.043). CONCLUSION: Intravenously administered dexamethasone appears to be more effective than intramuscularly administered betamethasone for the antepartum treatment of mothers with HELLP syndrome. PMID- 11408850 TI - Obstetrician-gynecologists performing genetic amniocentesis may be misleading themselves and their patients. AB - OBJECTIVE: Our purpose was to compare midtrimester amniocentesis-related fetal loss rates between obstetrician-gynecologists and perinatologists. STUDY DESIGN: This cohort study analyzes 1384 midtrimester amniocenteses from January 1, 1996, to December 31, 1999. Obstetrician-gynecologists who split their practices between two or more hospitals and explained fetal losses (eg, fetal anomalies, aneuploidy) were excluded from analysis. Eight obstetrician-gynecologists performed 138 procedures; 3 perinatologists performed 1246 procedures. Three experienced obstetrician-gynecologists accounted for 113 procedures. Analysis was by chi2. RESULTS: Within 30 days of midtrimester amniocentesis, there were 3 fetal losses for obstetrician-gynecologists and 4 for perinatologists (P =.02, chi2 = 5.19, degrees of freedom = 1). Obstetrician-gynecologist loss rates were 1 in 46 procedures versus 1 in 312 procedures for perinatologists. Losses were clustered among the 3 experienced obstetrician-gynecologists (P <.01, chi2 = 6.93, degrees of freedom = 1). The experienced obstetrician-gynecologist fetal loss rate was 1 in 38 amniocenteses, and the perinatologist fetal loss rate was 1 in 312. CONCLUSION: The risk of fetal loss from midtrimester amniocentesis appears to be higher when performed by an obstetrician-gynecologist compared with a perinatologist. PMID- 11408851 TI - Angiotensinogen and endothelial nitric oxide synthase gene polymorphisms among Hispanic patients with preeclampsia. AB - OBJECTIVE: We sought to establish an association between preeclampsia and the methionine to threonine polymorphism at amino acid residue 235 (Met235Thr) in angiotensinogen in a Hispanic population. We looked for a relationship between this allele and the allele in the endothelial nitric oxide synthase gene (NOS3) that produces the A form (NOS3*A) with respect to preeclampsia. STUDY DESIGN: Clinical data were collected from 87 patients with preeclampsia and 53 control subjects. Patients and controls were genotyped for the angiotensinogen polymorphism allele (AGT*T) and the NOS3*A polymorphism. We then compared patients with preeclampsia and control subjects and investigated disease severity within the preeclampsia group as a function of genotype. RESULTS: The AGT*T allelic frequencies among patients with preeclampsia and control subjects were 0.72 and 0.70, respectively (P =.84). The blood pressure of patients with an AGT*T allele who also carried a NOS3*A allele was higher at earlier gestational ages (r = -0.052; P =.02). Analysis suggested that the systolic blood pressure differences were due to gestational age effects and the presence of a NOS3*A allele (P <.10). CONCLUSION: The AGT*T allele was not associated with the development of preeclampsia. Independently of the presence of an AGT*T allele, the NOS3*A allele was associated with a higher blood pressure at an earlier gestational age. PMID- 11408852 TI - A randomized controlled trial comparing amoxicillin and azithromycin for the treatment of Chlamydia trachomatis in pregnancy. AB - OBJECTIVE: Our goal was to compare the efficacy of azithromycin with that of amoxicillin for the treatment of Chlamydia trachomatis infection during pregnancy. STUDY DESIGN: A randomized controlled trial of pregnant women with cervical C trachomatis infection receiving care at two inner-city, university based prenatal clinics. Pregnant women were randomly assigned to receive either oral amoxicillin, 500 mg, three times daily for 7 days, or oral azithromycin, 1 g, in a single dose. Partners were referred for treatment. Tests of cure were scheduled 4 weeks after initiation of treatment. Statistical analysis was performed by using the Student t test and chi2 analysis. RESULTS: One hundred twenty-nine pregnant women were enrolled, and 110 (85%) completed the protocol. There was similar treatment efficacy between amoxicillin and azithromycin (58% vs 64%, respectively,P =.56). In the amoxicillin group 3 women (5.5%) were intolerant, compared with 6 (10.9%) in the azithromycin group (P =.31). CONCLUSION: Amoxicillin and azithromycin are equally efficacious in the treatment of cervical C trachomatis during pregnancy. PMID- 11408853 TI - Prospective randomized trial of polyglactin 910 mesh to prevent recurrence of cystoceles and rectoceles. AB - OBJECTIVE: Our aim was to evaluate the efficacy of polyglactin 910 mesh in preventing recurrent cystoceles and rectoceles. STUDY DESIGN: In a prospective, randomized, controlled trial, patients undergoing vaginal reconstructive surgery with cystoceles to the hymenal ring and beyond were randomly selected to undergo anterior and posterior colporrhaphy with or without polyglactin 910 mesh reinforcement. Results were evaluated preoperatively and at 2, 6, 12, and 52 weeks postoperatively. RESULTS: A total of 161 women were randomly selected for this study. One woman was excluded at the time of surgery, and 17 women were lost to follow-up. Eighty women received mesh, and 80 did not. Both groups were found to be equivalent with respect to age, parity, concomitant surgery, and menopausal and hormone replacement status. Preoperatively 49 women had a central cystocele to the hymenal ring and 111 women had cystoceles beyond the introitus; 91 women had a rectocele to the mid-vaginal plane, 31 to the hymenal ring, and 22 beyond the introitus. After 1 year, 30 (43%) of 70 subjects without mesh and 18 (25%) of 73 subjects with mesh had recurrent cystoceles beyond the mid-vaginal plane (P =.02). Eight women without mesh and 2 women with mesh had recurrent cystoceles to the hymenal ring (P =.04). No recurrent cystoceles beyond the hymenal ring occurred in either group. Multivariate logistic regression analysis showed concurrent slings to be associated with significantly fewer recurrent cystoceles (odds ratio, 0.32; P =.005), whereas the presence of mesh remained significantly predictive of fewer cystocele recurrences in this analysis. Thirteen recurrent rectoceles were noted 1 year postoperatively, with no differences between groups. CONCLUSION: Polyglactin 910 mesh was found to be useful in the prevention of recurrent cystoceles. PMID- 11408854 TI - Failed vaginal birth after a cesarean section: how risky is it? I. Maternal morbidity. AB - OBJECTIVE: Our purpose was to determine the maternal risks associated with failed attempt at vaginal birth after cesarean compared with elective repeat cesarean delivery or successful vaginal birth after cesarean. STUDY DESIGN: From 1989 to 1998 all patients attempting vaginal birth after cesarean and all patients undergoing repeat cesarean deliveries were reviewed. Data were extracted from a computerized obstetric database and from medical charts. The following three groups were defined: women who had successful vaginal birth after cesarean, women who had failed vaginal birth after cesarean, and women who underwent elective repeat cesarean. Criteria for the elective repeat cesarean group included no more than two previous low transverse or vertical incisions, fetus in cephalic or breech presentation, no previous uterine surgery, no active herpes, and adequate pelvis. Predictor variables included age, parity, type and number of previous incisions, reasons for repeat cesarean delivery, gestational age, and infant weight. Outcome variables included uterine rupture or dehiscence, hemorrhage >1000 mL, hemorrhage >2000 mL, need for transfusion, chorioamnionitis, endometritis, and length of hospital stay. The Student t test and the chi(2) test were used to compare categoric variables and means; maternal complications and factors associated with successful vaginal birth after cesarean were analyzed with multivariate logistic regression, allowing odds ratios, adjusted odds ratios, 95% confidence intervals, and P values to be calculated. RESULTS: A total of 29,255 patients were delivered during the study period, with 2450 having previously had cesarean delivery. Repeat cesarean deliveries were performed in 1461 women (5.0%), and 989 successful vaginal births after cesarean delivery occurred (3.4%). Charts were reviewed for 97.6% of all women who underwent repeat cesarean delivery and for 93% of all women who had vaginal birth after cesarean. Vaginal birth after cesarean was attempted by 1344 patients or 75% of all appropriate candidates. Vaginal birth after cesarean was successful in 921 women (69%) and unsuccessful in 424 women. Four hundred fifty-one patients undergoing cesarean delivery were deemed appropriate for vaginal birth after cesarean. Multiple gestations were excluded from analysis. Final groups included 431 repeat cesarean deliveries and 1324 attempted vaginal births after cesarean; in the latter group 908 were successful and 416 failed. The overall rate of uterine disruption was 1.1% of all women attempting labor; the rate of true rupture was 0.8%; and the rate of hysterectomy was 0.5%. Blood loss was lower (odds ratio, 0.5%; 95% confidence interval, 0.3-0.9) and chorioamnionitis was higher (odds ratio, 3.8%; 95% confidence interval, 2.3-6.4) in women who attempted vaginal births after cesarean. Compared with women who had successful vaginal births after cesarean, women who experienced failed vaginal births after cesarean had a rate of uterine rupture that was 8.9% (95% confidence interval, 1.9-42) higher, a rate of transfusion that was 3.9% (95% confidence interval, 1.1-13.3) higher, a rate of chorioamnionitis that was 1.5% (95% confidence interval, 1.1-2.1) higher, and a rate of endometritis that was 6.4% (95% confidence interval, 4.1-9.8) higher. CONCLUSION: Patients who experience failed vaginal birth after cesarean have higher risks of uterine disruption and infectious morbidity compared with patients who have successful vaginal birth after cesarean or elective repeat cesarean delivery. Because actual numbers of morbid events are small, caution should be exercised in interpreting results and counseling patients. More accurate prediction for safe, successful vaginal birth after cesarean delivery is needed. PMID- 11408855 TI - Inhibition of cervical ripening by local application of cyclooxygenase 2 inhibitor. AB - OBJECTIVE: Our goal was to test the hypothesis that the previously demonstrated progesterone-independent prolongation of pregnancy in rats treated with cervical application of the cyclooxygenase 2 inhibitor nimesulide is the result of inhibition of cervical ripening. STUDY DESIGN: Timed-pregnant Sprague-Dawley rats were randomly allocated to treatment with 50 mg nimesulide or vehicle, applied daily on the cervix for 5 days (days 14-18). On day 19 the animals were humanely killed and the cervices were removed. In the first series of experiments the cervices of animals treated with nimesulide (n = 10) or vehicle (n = 10) were examined with a cervimeter, which stretches the cervical tissues in incremental steps of 0.2 mm at 1-minute intervals. A steeper slope through the linear portion of the resulting force-versus-displacement curve indicates more resistance to stretch. In the second series of experiments the cervices of animals treated with nimesulide (n = 11) or vehicle (n = 11) were examined with the Collascope optical device. The cervical content of cross-linked collagen was measured with light induced fluorescence. The fluorescence spectrum at 390 nm (peak wavelength of the collagen spectrum) was determined. For standardization, the ratio of counts of collagen peak over reference counts was used in the final analyses as an indicator of cross-linked collagen content. RESULTS: Animals treated with cervical application of nimesulide had significantly higher resistance to stretch than controls (slope: 0.2564 +/- 0.1213 vs 0.1387 +/- 0.0652; P =.019). The cervical content of cross-linked collagen was not significantly different between nimesulide-treated animals and controls (light-induced fluorescence ratios: 3.2134 +/- 0.7390 vs 2.7892 +/- 0.8518; P =.227). CONCLUSIONS: Treatment with cervical application of the cyclooxygenase 2 inhibitor nimesulide prevents the physiologic process of cervical ripening in late pregnancy. The inhibition is not the result of changes in cross-linked collagen content. Inhibition of cervical ripening with locally administered cyclooxygenase 2 inhibitor may be a potentially valuable treatment for patients at risk for preterm delivery. PMID- 11408856 TI - Analysis of surgical complications and patient outcomes in a residency training program. AB - OBJECTIVES: This study compares surgical complications and patient outcomes between pelvic reconstructive surgery performed by an experienced surgeon (group 1) and those performed by resident physicians with the senior surgeon assisting and teaching (group 2). STUDY DESIGN: During a 5-year interval, 310 consecutive women underwent vaginal prolapse repair. Demographic, historic, and preoperative physical examination variables were compared. Intraoperative and postoperative outcomes were also compared. RESULTS: Patients operated on by the senior surgeon (Bob L. Shull) were thinner (group 1 vs group 2: 25.8 kg/m2 vs 27.1 kg/m2; P =.014), more often had prior prolapse or incontinence procedures (55% vs 33%; P <.001), and required shorter operating times (124 minutes vs 140 minutes; P =.002). The senior surgeon's patients differed from the resident physicians' patients with regard to stage of pelvic organ prolapse. No differences were observed for patient age (P =.51), estimated blood loss (P =.50), urologic complications (P =.59), and hospital stay (P =.25). The durability of the repairs was not different between the groups. CONCLUSIONS: We have demonstrated that in a tertiary referral practice resident surgeons can be taught to perform complex vaginal surgery with the only observed disadvantage being a slightly prolonged operative time. PMID- 11408857 TI - Challenging generally accepted contraindications to vaginal hysterectomy. AB - OBJECTIVE: A number of preexisting clinical conditions are generally accepted as contraindications to vaginal hysterectomy. The purpose of this study was to evaluate the validity of this concept. STUDY DESIGN: The study vaginal hysterectomy group consisted of 250 consecutive patients undergoing vaginal hysterectomy. These patients (1) had a large uterus (>180 g), (2) either were nulliparous or had no previous vaginal delivery, or (3) had a previous cesarean delivery or pelvic laparotomy. Three control groups used for comparison underwent (1) laparoscopically assisted vaginal hysterectomy, (2) vaginal hysterectomy, or (3) abdominal hysterectomy. The records for all patients were analyzed for age, weight, parity, primary diagnosis, uterine size, operative time, blood loss, analgesia, hospital stay, resumption of diet, incidence of morcellation, and surgical complications. Sample size calculations were based on previous studies of complications associated with vaginal hysterectomy (alpha =.05; beta =.20). RESULTS: Hysterectomy was successfully completed by the intended vaginal route in all study patients. Major and minor complications (3.2%) were significantly less (P <.001) than in the other groups as follows: vaginal hysterectomy, 10.4%; laparoscopically assisted vaginal hysterectomy, 11.6%; and abdominal hysterectomy, 13.6%. The decrease in hematocrit was 5.7% in the study vaginal hysterectomy group compared with 6.2% for vaginal hysterectomy, 6.5% for abdominal hysterectomy (P =.009), and 6.6% for laparoscopically assisted vaginal hysterectomy (P =.002). Hospital stay was shorter for the study group (2.1 days) than for vaginal hysterectomy (2.3 days; P <.001) and abdominal hysterectomy (2.7 days; P <.001). Operative time was shorter in the study vaginal hysterectomy group (49 minutes) than with laparoscopically assisted vaginal hysterectomy (76 minutes; P <.001) or abdominal hysterectomy (61 minutes; P <.001), although morcellation was carried out more frequently in the study group (34%) than with vaginal hysterectomy (4%) or laparoscopically assisted vaginal hysterectomy (11%). CONCLUSION: Our data indicate that a large uterus, nulliparity, previous cesarean delivery, and pelvic laparotomy rarely constitute contraindications to vaginal hysterectomy. PMID- 11408858 TI - Support for an infection-induced apoptotic pathway in human fetal membranes. AB - OBJECTIVE: Lipopolysaccharide and tumor necrosis factor alpha levels are both elevated in the amniotic fluid of women during infection-associated preterm labor and premature rupture of fetal membranes. Our laboratory has shown that apoptosis is associated with premature rupture of fetal membranes but is not associated with preterm labor. The exact pathway that leads to apoptosis-mediated premature rupture of fetal membranes is still unclear. Because infection and increased inflammatory cytokine response are associated with the majority of cases of premature rupture of fetal membranes, we examined the roles of bacterial lipopolysaccharide and tumor necrosis factor alpha in inducing the proapoptotic caspase pathway in fetal membranes. STUDY DESIGN: Amniochorionic membranes collected from women undergoing elective repeat cesarean delivery at term were placed in an organ explant system. At the end of a 48-hour incubation period, membranes were stimulated with lipopolysaccharide (50 ng/mL) and recombinant tumor necrosis factor (50 ng/mL). Total ribonucleic acid extracted from these samples was subjected to reverse transcription and two separate sets of multiple polymerase chain reaction. One set studied the expression of Fas, Fas ligand, caspase 8, Fas-associated death domain, and tumor necrosis factor receptor associated death domain genes and the second set studied the expression of caspase 2, 4, 6, 7, and 10. Caspase 2, 3, and 9 expression was also studied by reverse transcriptase-polymerase chain reaction. RESULTS: Multiple polymerase chain reactions and reverse transcriptase-polymerase chain reactions documented the induction of Fas and caspase 2, 3, 7, 8, and 9 genes in amniochorion after lipopolysaccharide and tumor necrosis factor stimulation compared with the nonstimulated controls. Neither lipopolysaccharide nor tumor necrosis factor induced Fas ligand expression in human fetal membranes. Caspase 3, 4, and 6, Fas associated death domain, and tumor necrosis factor receptor-associated death domain expressions were constitutive in all the tissues tested; however, tumor necrosis factor receptor-associated death domain expression appeared stronger in tumor necrosis factor-stimulated tissues. CONCLUSION: The presence of the signal docking proteins tumor necrosis factor receptor-associated death domain and Fas associated death domain and the induction of caspase cascade initiators (caspase 2, 8, and 10) and effector caspases (caspase 3, 6, 7, and 9) by lipopolysaccharide and tumor necrosis factor suggest that tumor necrosis factor tumor necrosis factor receptor-mediated apoptosis may occur in the human fetal membrane. PMID- 11408859 TI - Distinct molecular events suggest different pathways for preterm labor and premature rupture of membranes. AB - OBJECTIVE: On a clinical level, the etiologies associated with premature rupture of the membranes and preterm labor are virtually identical, though these conditions end in distinctly different events. This study was designed to determine differences between preterm labor and preterm premature rupture of membranes by using molecular markers of extracellular matrix degradation and apoptosis. STUDY DESIGN: Amniochorion and amniotic fluid samples were collected from gestational age-matched groups of women undergoing cesarean delivery before term. Samples were collected from 2 groups of women, women with premature rupture of membranes and women with preterm labor with no rupture of membranes. Changes in the expression pattern of messenger ribonucleic acid for matrix metalloproteinases (MMP), tissue inhibitor of metalloproteinases (TIMP), and pro apoptotic (p53 and Bax) and anti-apoptotic (Bcl-2) proteins were identified by quantitative polymerase chain reaction. Enzyme-linked immunosorbent assay was used to determine the levels of these proteins in the amniotic fluid. Multiplex polymerase chain reaction was performed to study the expression of Fas-Fas ligand associated pro-apoptotic genes. Unpaired nonparametric, 2-tailed Mann-Whitney U test was used to determine statistical significance of quantitative polymerase chain reaction and enzyme-linked immunosorbent assay (P <.05 was considered significant). RESULTS: Quantitative polymerase chain reaction results demonstrated an increased mRNA expression for MMP2, MMP9, and MT1-MMP and a decreased expression for TIMP2 in prematurely ruptured membranes compared with preterm labor membranes. Enzyme-linked immunosorbent assay documented increases in the amniotic fluid concentrations of immunoreactive and bioactive MMP2 and MMP9 and immunoreactive MMP3 and a decreased TIMP2 concentration in fluids obtained from the premature rupture of membranes group compared with the preterm labor group. The pro-apoptotic genes p53 and bax were up-regulated in premature rupture of membranes when compared with preterm labor. Anti-apoptotic gene (Bcl-2 ) expression was increased in preterm labor membranes compared with prematurely ruptured membranes. Interleukin-18 (a pro-apoptotic cytokine) was increased in the amniotic fluid during premature rupture of membranes compared with preterm labor. Prematurely ruptured membranes also demonstrated fragmented deoxyribonucleic acid and expression of Fas and caspase 8 (apoptosis initiator), which were all absent in preterm labor membranes. CONCLUSIONS: We have begun to delineate 2 divergent molecular pathways for premature rupture of membranes and preterm labor. Most likely, this is the beginning of the identification of differences that will become evident with the use of molecular biology. PMID- 11408860 TI - Endometriosis: correlation between histologic and visual findings at laparoscopy. AB - OBJECTIVE: The purpose of this study was to correlate the diagnosis of endometriosis on the basis of visualization at laparoscopy with the pathologic diagnosis. STUDY DESIGN: A prospective study of 44 patients undergoing laparoscopy for the evaluation of chronic pelvic pain was carried out. All areas suggestive of endometriosis were excised and examined pathologically. Peritoneal biopsy specimens were obtained from areas of normal-appearing peritoneum to rule out microscopic endometriosis. All lesions were identified by anatomic site. Visual and histologic American Fertility Society scores were compared. The positive predictive value, sensitivity, negative predictive value, and specificity were determined for visually identified endometriosis versus the histologic correlate. RESULTS: The mean prevalence of abnormalities visually consistent with endometriosis was 36%, with 18% confirmed histologically. The positive predictive value was 45%; sensitivity, 97%; negative predictive value, 99%; and specificity, 77%; for visual versus histologic diagnosis of endometriosis. Thirty-six percent of the diagnoses were downstaged on the basis of histologic findings. CONCLUSION: A diagnosis of endometriosis should be established only after histologic confirmation. PMID- 11408861 TI - Obstetric ultrasonographic findings and fetal chromosomal abnormalities: refining the association. AB - OBJECTIVE: In an attempt to refine the role of ultrasonography in screening and identifying fetuses at risk for chromosomal abnormalities, a retrospective review of patients undergoing genetic amniocentesis was undertaken. STUDY DESIGN: Computer databases from the perinatal biology laboratory and cytogenetics laboratory of our institution were correlated to compare the results of the fetus' ultrasonographic examination with the cytogenetic results from amniocentesis. Univariate and multivariate analysis were used to determine the best correlations between ultrasonographic findings and chromosomal abnormalities (study 1). The results were used to construct regression analysis formulas and a Neural Network program to predict the presence or absence of chromosomal abnormalities in a second set of patients (study 2) undergoing genetic amniocentesis. RESULTS: One hundred twenty-five chromosomal abnormalities were found in 3775 fetuses in study 1 (3.3%). Multivariate analysis showed significant correlations between anomalies of the central nervous system, heart, face and neck, and extremities and increased nuchal fold, increased bowel echogenicity, abnormal biparietal diameter-to-femur ratio, and the presence of chromosomal abnormalities. Regression equations and a Neural Network program successfully predicted the presence or absence of fetal chromosomal abnormalities in a second set of 901 at-risk fetuses. CONCLUSION: A normal ultrasonographic examination result in patients who are at increased risk for fetal chromosomal abnormalities reduces the risk 2- to 3-fold, whereas the presence of any major ultrasonographic abnormality or certain minor abnormalities significantly increases the risk. The application of these results to low-risk patients is still premature. PMID- 11408862 TI - Meconium aspiration syndrome in term neonates with normal acid-base status at delivery: is it different? AB - OBJECTIVE: Our aim was to compare the clinical characteristics of meconium aspiration syndrome in cases with pH > or =7.20 and in those with pH <7.20. STUDY DESIGN: Medical records of diagnostic codes from the International Classification of Diseases, Ninth Revision, were used to identify neonates with severe meconium aspiration syndrome who had been delivered at our institution from 1994 through 1998. Severe meconium aspiration syndrome was defined as a mechanical ventilator requirement of >48 hours. Clinical data including neonatal outcomes of cases of meconium aspiration syndrome associated with umbilical pH > or =7.20 at delivery were compared with data on outcomes of cases with pH <7.20. RESULTS: During this 4-year study period, 4985 singleton term neonates were delivered through meconium stained amniotic fluid. Forty-eight cases met all study criteria, and pH values at delivery were as follows: pH > or =7.20, n = 29, and pH <7.20, n = 19. There were no differences between groups in the incidence of clinical chorioamnionitis, in the presence of meconium below the vocal cords, or in birth weight. Neonates with meconium aspiration syndrome and umbilical pH > or =7.20 at delivery developed seizures as often as those with pH <7.20 (20.1% vs 21.1%; P = 1.0). CONCLUSION: Normal acid-base status at delivery is present in many cases of severe meconium aspiration syndrome, which suggests that either a preexisting injury or a nonhypoxic mechanism is often involved. PMID- 11408863 TI - How to predict recurrent shoulder dystocia. AB - OBJECTIVE: Our aim was to determine the rate and risk factors for recurrent shoulder dystocia. STUDY DESIGN: A retrospective analysis of patients diagnosed with shoulder dystocia was performed by searching a computerized database from January 1, 1993, to June 30, 1999 for the following information: (1) vaginal deliveries, either spontaneous or operative, (2) shoulder dystocia, (3) birth weight, (4) duration of second stage of labor, (5) parity, and (6) gestational diabetes. Statistical analyses included chi(2) and t test. RESULTS: There were 39,681 vaginal deliveries with 602 (1.5%) complicated by shoulder dystocia. Sixty six patients underwent a subsequent vaginal delivery, and 11 (16.7%) experienced another shoulder dystocia. The odds ratio for a recurrent shoulder dystocia was 10.98 (P <.000001). Nine of the 11 patients with recurrent shoulder dystocia compared with 28 of 55 without a recurrence were nulliparous women in their index pregnancy (P <.001). The mean fetal weights were 3885 g in the recurrent dystocia group and 3702 g in the group without recurrence (P <.03). Gestational age, operative delivery, and gestational diabetes were similar in the two groups. CONCLUSION: Factors that appear to increase the recurrence risk of shoulder dystocia include fetal weight and maternal parity. Prior shoulder dystocia is the single greatest predictive factor. PMID- 11408864 TI - A new device for measuring intrauterine temperature. AB - OBJECTIVE: We evaluated a new device that uses the intrauterine pressure catheter to measure the maternal temperature in patients who are in labor. STUDY DESIGN: The study was conducted at two medical centers, Christiana Hospital in Newark, Delaware, and Saint Louis University/St Mary's Health Center in St Louis, Missouri, from September 1, 1997, to May 2, 1998. An intrauterine pressure catheter with a thermistor sensor in the tip was placed into the uterus after spontaneous rupture of membranes. The intrauterine, oral, and tympanic temperatures were simultaneously obtained immediately after insertion of the intrauterine pressure catheter and then hourly until delivery or the initiation of amnioinfusion. RESULTS: The study comprised 97 patients and 404 temperature readings with a temperature range of 34.7 degrees C to 40.7 degrees C. The normal mean +/- SD for the oral, tympanic, and intrauterine temperatures was 36.7 degrees C +/- 0.5 degrees C, 36.8 degrees C +/- 0.5 degrees C, and 37.3 degrees C +/- 0.4 degrees C, respectively. There was a linear relationship among the oral, tympanic, and intrauterine temperatures. All three methods showed a significant increase in mean body temperature after epidural anesthesia. CONCLUSION: The new device, the intrauterine pressure-temperature catheter, provides a convenient and accurate means of continuously measuring uterine temperature in patients who are in labor and require intrauterine monitoring. PMID- 11408865 TI - Who will lead medicine in the new millennium? PMID- 11408866 TI - Expression of human telomerase reverse transcriptase, the catalytic subunit of telomerase, is associated with the development of persistent disease in complete hydatidiform moles. AB - OBJECTIVE: This study was undertaken to determine the putative role of telomerase activity and human telomerase reverse transcriptase (hTERT) expression in the development of persistent disease in patients with a diagnosis of complete hydatidiform mole. The ribonucleoprotein telomerase has been shown to have a major role in the process of cellular immortality and carcinogenesis. The reactivation of this enzyme that occurs in the development of malignancies appears to be limited by the regulation of its catalytic subunit hTERT. Compared with their somatic counterparts, most human malignancies demonstrate telomerase activity, and this activity is dependent on the cellular presence of hTERT. The role of telomerase in the pathogenesis of complete hydatidiform moles is not clearly understood. Moreover, the role of hTERT in trophoblastic disease, as well as in the development of persistent trophoblastic disease, has yet to be elucidated. STUDY DESIGN: Telomerase activity and hTERT expression were analyzed in the initial uterine evacuation specimen of 54 complete hydatidiform moles by use of the telomeric repeat amplification protocol assay and reverse transcription-polymerase chain reaction methods. The results were compared and then correlated with the development of persistent trophoblastic disease. RESULTS: Among the 54 patients who were examined with a diagnosis of complete hydatidiform mole, persistent trophoblastic disease requiring postevacuation chemotherapy developed in 6. In the remaining 48 patients, spontaneous remission of the disease occurred after uterine evacuation. Both telomerase activity and hTERT expression were detected in all 6 cases of persistent disease on the initial molar tissue sampled. Among the 48 nonpersistent moles, telomerase activity was detected in 29 (60%) and hTERT expression was demonstrated in 26 (54%). The detection of hTERT expression was significantly associated with the presence of persistent disease (P =.035). Moreover, the absence of hTERT expression in molar tissue obtained from uterine evacuation demonstrated a 100% negative predictability in determining cases of complete mole that were nonpersistent. CONCLUSIONS: Compared with telomerase activity, the expression of hTERT is significantly associated with the development of persistent disease in complete hydatidiform moles. The absence of hTERT expression in the initial tissue sample from complete moles may have potential clinical value in determining patients who will eventually undergo spontaneous remission after uterine evacuation. PMID- 11408867 TI - Cervical cerclage in the second trimester of pregnancy: a historical cohort study. AB - OBJECTIVE: The purpose of this study was to compare second-trimester transvaginal cervical cerclage with conservative management on duration of pregnancy and perinatal outcome in patients with early or advanced cervical changes. STUDY DESIGN: A historical cohort analysis was performed. Maternal and neonatal records between 1995 and 1999 were retrospectively reviewed for women presenting between 18 and 27 weeks of gestation with early cervical changes (length <3 cm, dilatation <2 cm, funneling of fetal membranes shown by transvaginal ultrasonography) (group 1, n = 31) and for women with advanced cervical effacement and dilatation (cervical dilatation > or =2 cm but < or =5 cm, fetal membranes visible) (group 2, n = 39). In each group, patients who underwent Shirodkar or McDonald cerclage were compared with patients treated conservatively with bed rest. Both groups also received multifactorial treatment with tocolytic agents, broad-spectrum antibiotics, and indomethacin. Outcome variables were analyzed for statistical significance by parametric and nonparametric methods. RESULTS: Regardless of treatment method, patients with early cervical changes (group 1) were given a diagnosis earlier and delivered later in pregnancy compared with their counterparts who had advanced cervical changes (group 2) (P <.05). In both patients who underwent cerclage and those treated conservatively, the mean birth weight among surviving infants was higher and the mean neonatal intensive care unit stay was shorter in group 1 than in group 2 (P <.02). However, duration of maternal hospital stay and neonatal survival rates were not different. In both groups 1 and 2, the interval from treatment to delivery, the mean gestational age at delivery, and mean birth weight were increased, whereas neonatal intensive care unit stay was decreased by cerclage treatment (P <.05). In group 1, a higher percentage of patients treated with cerclage received antibiotics and indomethacin than did control subjects (P <.01), whereas in group 2, the use of multifactorial treatment was not different (P =.5). The duration of maternal hospital stay and neonatal survival did not differ significantly among patients treated conservatively or with cerclage. CONCLUSIONS: Diagnosis of premature cervical changes by ultrasonography was correlated with treatment earlier in gestation and with a favorable impact on perinatal outcome in both patients treated with cerclage and those treated conservatively. Cervical cerclage was associated with an improved perinatal outcome (in comparison with conservative therapy) in women with early cervical changes detected by ultrasonography and in patients with advanced cervical dilatation and visible membranes. However, the apparent therapeutic effect of cerclage in patients with mild cervical incompetence may be due in part to an increased use of antibiotics and indomethacin in conjunction with cerclage. PMID- 11408868 TI - Evaluation of atypical glandular cells of undetermined significance: is age important? AB - OBJECTIVE: Our purpose was to assess the effect of age on the incidence of significant pathologic findings in patients with atypical glandular cells of undetermined significance. STUDY DESIGN: This retrospective study evaluated nonreferred patients with Papanicolaou smears showing atypical glandular cells of undetermined significance over a 5-year period. RESULTS: Two hundred eighty-one women (91%) with Papanicolaou smears showing atypical glandular cells of undetermined significance were available for follow-up. Significant abnormality was found in 90 subjects (32%), including 31 women (11%) with cancer. Older patients (> or =50 years old) have lower odds of high-grade cervical dysplasia (odds ratio = 0.21; 95% CI: 0.32-0.40) and higher odds of uterine cancer (odds ratio = 12.88; 95% CI: 1.78-567.60) compared with younger patients. CONCLUSIONS: Women with Papanicolaou smears showing atypical glandular cells of undetermined significance have a 1 in 3 chance of having significant abnormality. Evaluation includes colposcopy, endocervical curettage, and appropriate cervical biopsies. Patients >45 years require endometrial biopsies. PMID- 11408869 TI - Testing surgical skills of obstetric and gynecologic residents in a bench laboratory setting: validity and reliability. AB - OBJECTIVE: Resident surgical skills are acquired mainly through observing and later performing procedures in the operating room. Evaluation of surgical skills has traditionally been done through subjective faculty evaluation, a technique that has poor reliability and unknown validity. Our goal was to develop specific surgical tasks, both laparoscopic and open abdominal, that could be objectively and reliably evaluated in a bench laboratory setting. STUDY DESIGN: The prospective development of a reliable and valid resident surgical skills test in a bench laboratory setting was our goal. A written test of surgical knowledge and 12 skills tests were administered to 36 residents. Laparoscopic bench tasks were simulated with the use of a box and camera with a video display. Six laparoscopic tasks were assessed, including placing pegs on a board, running the bowel simulation, and other tasks that involve hand-eye coordination and manual dexterity. Open abdominal skills simulated incision closure, suturing a vaginal cuff, knot tying, and using a tie on a passer. Residents were timed at each given station and were given a rating score by 2 examiners. RESULTS: Knowledge scores showed a significant improvement by residency level. Assessment of construct validity (the ability to discriminate among residency levels) demonstrated significant differences on the rating of overall performance and individual tasks by level (determined by 1-way analysis of variance). Interrater reliability (agreement between 2 raters) with the use of intraclass correlation was 0.79 for the total score. The cost to administer the bench laboratory test was less than $50 and required 30 hours of faculty time. CONCLUSION: The results of this study suggest that surgical bench laboratory tasks can assess residents' surgical skills with good reliability and validity on most tasks. Our previous study, which used an animal laboratory, was expensive, and the bench laboratory model may provide an alternative means to assess surgical skills. PMID- 11408870 TI - Multiple gestations in assisted reproductive technology: can they be avoided with blastocyst transfers? AB - OBJECTIVE: Blastocysts are advanced-stage embryos with high implantation potential; theoretically, limited numbers of blastocysts can be used for embryo transfer to achieve good pregnancy rates with low multiple pregnancy rates. Clinical outcomes of a newly implemented blastocyst transfer program were evaluated. STUDY DESIGN: This study is a retrospective analysis of 553 blastocyst transfer cycles performed by a university-based in vitro fertilization program; risk factors associated with multiple gestations were analyzed. RESULTS: An average of 2.2 embryos were used for embryo transfer. The overall clinical pregnancy rate per embryo transfer was 45.1%; multiple gestation, twin, and triplet rates were 40.9%, 36.5%, and 4.3%, respectively. Multiple gestations increased significantly (1) when embryo transfer was done on day 5, (2) when > or =2 blastocysts were present on day 5, and (3) when maternal age was < or =30 years. CONCLUSION: In spite of a conservative approach to the number of blastocysts used for embryo transfer, the overall multiple pregnancy rate was high, and triplet pregnancies did occur. PMID- 11408871 TI - Is vaginal birth after cesarean safe? Experience at a community hospital. AB - OBJECTIVE: We sought to evaluate the effectiveness and safety of promoting a trial of labor after prior cesarean birth in a community hospital. STUDY DESIGN: A 4-year prospective cohort study was conducted of all patients who had prior cesarean births (N = 1481). A comparison of outcomes was performed between those who elected repeat cesarean delivery (n = 727) and those who attempted a trial of labor after previous cesarean(s) (n = 754). RESULTS: We found that the vaginal birth after cesarean attempt rate was 50.9% and declined significantly during the last 2 years of the study. The elective repeat cesarean rate was 49.1% and increased significantly during the last 2 years of the study. In addition, we found that neonatal outcomes were similar, with the exception of 2 neonatal deaths caused by uterine rupture. Twelve uterine ruptures occurred (1.6%), and 11 of the 12 ruptures involved either induction or augmentation of labor, or both. CONCLUSIONS: A trial of vaginal birth after cesarean is safe provided that induction of labor is not used. The uterine rupture rate of 1.6% is higher than reported in the literature; this may reflect underreporting by community hospitals. PMID- 11408872 TI - Rectal prolapse in women with other defects of pelvic floor support. AB - OBJECTIVES: We describe a series of patients with rectal prolapse who had other pelvic floor defects. STUDY DESIGN: Patients with rectal prolapse that we examined between 1990 and 2000 were reviewed. RESULTS: During this time frame 55 patients with rectal prolapse were seen by one of us. Fifty-two of these patients had other defects of pelvic floor support and are the subject of this report. The diagnosis was established in all patients with video defecography. Thirty-nine of the patients had internal (occult) prolapse that simulated either a rectocele or an enterocele. The mean number of surgical procedures for pelvic floor support before the diagnosis of rectal prolapse was 1.5. Thirty-one patients underwent a sigmoid resection with rectopexy, 12 underwent a rectopexy alone, 3 underwent a Ripstein procedure, 2 elderly patients had physical therapy alone, and the other 4 patients had surgical correction of the rectal prolapse before being referred for repair of vaginal vault prolapse. Other procedures performed simultaneously included sacral colpopexy, sacrospinous suspension, rectopubic urethropexy, and abdominal fixation of the vagina to the uterosacral ligaments. CONCLUSIONS: Rectal prolapse frequently coexists with other pelvic floor defects. Internal rectal prolapse may simulate a rectocele or enterocele and requires defecography to establish the diagnosis. Rectopexy (with or without sigmoid resection) is a satisfactory technique for correction and may be combined with other reconstructive procedures on the pelvic floor. PMID- 11408873 TI - The demographics of pelvic floor disorders: current observations and future projections. AB - OBJECTIVE: Our aim was to assess current demand for care of pelvic floor disorders and create projections for future demand for care. We also sought to better understand the characteristics of women seeking care. STUDY DESIGN: Current demand for care was calculated by comparing those women seeking care through the female pelvic floor disorders clinic with those women of the same age range at risk within an integrated health care delivery program. Patients underwent complete urogynecologic evaluation including cystometry. Women seeking care were compared with regard to age, distribution of conditions (pelvic organ prolapse, stress conditions, urge conditions), and probability of undergoing surgery. Modeling the study population by use of data from the US Census Bureau, which projects population changes over the next 30 years, created predictions of future demand. RESULTS: Data were available on 2070 consecutive patients with an age range of 30 to 89 years normally distributed around a median age of 61.5 years drawn from an at-risk population of 149,000 women aged 30 to 89 years. Older women generated more consults per 1000 woman years than did the younger cohorts (1.7 vs 18.6 consults per 1000 woman years for those 30-39 years old vs those 70-79 years old; P <.05). Estimates of growth in demand at 30 years indicate a 45% increase in demand while net growth of the same population segment should be 22%. Stress conditions were more common among younger women and urge conditions were more common among older women. Pelvic organ prolapse was equally distributed throughout the age ranges. CONCLUSIONS: Over the next 30 years, growth in demand for services to care for female pelvic floor disorders will increase at twice the rate of growth of the same population. Demand for care for pelvic floor disorders comes from a wide age range of women, although mature age groups generate 10 times the number of consults per 1000 woman years as do their younger counterparts. Age plays a major role in the distribution of conditions with which patients present. These findings have broad implications for those responsible for administering programs to care for women, allocating research funds in women's health and geriatrics, and training physicians to meet this rapidly escalating demand. PMID- 11408874 TI - Obstetrical deliveries associated with maternal malignancy in California, 1992 through 1997. AB - OBJECTIVE: This study aims to characterize the rate of occurrence and nature of outcomes associated with obstetrical deliveries in women with malignant neoplasms among 3,168,911 women who delivered in California in 1992 through 1997. DESIGN: The study is a population-based retrospective review of infant birth and death certificates and maternal and neonatal discharge records. Cases of malignant neoplasms associated with obstetrical delivery were attributed to 1 of 3 categories, depending on the earliest documented hospital discharge diagnosis, as follows: "prenatal" if the diagnosis was first documented by hospitalization within 9 months preceding delivery, "at delivery" if the diagnosis was established from the delivery hospitalization, or "postpartum" if the diagnosis was first documented by hospitalization within 12 months after delivery. METHODS: Computer-linked infant birth and death certificates and maternal and neonatal discharge records were used to identify cases and outcomes. Cases of malignant neoplasms were identified by using International Classification of Diseases, Ninth Revision codes (140-208). Noninvasive neoplasms and carcinoma in situ neoplasms were excluded. In analysis of outcomes, the Mantel-Haenszel estimate for adjusted odds ratios was used. RESULTS: Among 3,168,911 obstetrical deliveries over the 6-year span, a total of 2247 cases of primary malignancy were identified. The observed rate of occurrence for primary malignant neoplasms was 0.71 per 1000 live singleton births. Most cases (53.3%) were first documented in the postpartum period as follows: prenatal, 587 cases (0.18 per 1000); at delivery, 462 cases (0.15 per 1000); and postpartum, 1198 cases (0.38 per 1000). The most frequently documented primary malignant neoplasms associated with obstetrical delivery were breast cancer (423 cases, 0.13 per 1000), thyroid cancer (389 cases, 0.12 per 1000), cervical cancer (266 cases, 0.08 per 1000), Hodgkin's disease (172 cases, 0.05 per 1000), and ovarian cancer (123 cases, 0.04 per 1000). Odds ratios for a variety of demographic factors identified maternal age as the most significant risk factor for development of malignant neoplasms (age greater than 40 vs 20-25, odds ratio 5.7, CI 4.6-6.9). Age-adjusted odds ratios for maternal cancer of any type suggested significantly elevated risks for cesarean delivery (odds ratio 1.4, CI 1.3-1.6), blood transfusion (odds ratio 6.2, CI 4.5-8.5), hysterectomy (odds ratio 27.4, CI 20.8-36.1), and maternal postpartum hospital stay greater than 5 days (odds ratio 30.6, CI 27.9-33.6), but not for postpartum maternal death (odds ratio 0.8, CI 0.6-1.0). Odds ratios also suggested significantly elevated risks for premature newborn (odds ratio 2.0, CI 1.8-2.2), very low birth weight (odds ratio 2.9, CI 2.2-3.8), and newborn hospital stay longer than 5 days (odds ratio 2.6, CI 2.4-3.0), but not for neonatal death (odds ratio 1.6, CI 0.8-3.1) or infant death (odds ratio 1.2, CI 0.5-3.3). However, several types of malignant neoplasms did confer significant elevations in risk for neonatal death. Hospital charges for both maternal and neonatal care were significantly elevated in the maternal malignant neoplasm group. CONCLUSION: A lower than expected occurrence rate of obstetrical delivery associated with maternal malignancy was seen when compared with previously published hospital-based reports. Malignant neoplasms associated with obstetrical delivery were most frequently first documented in the postpartum period. Maternal and neonatal morbidity were significantly increased, yet the risk of in-hospital maternal death was not significantly elevated. A significant increase in risk of neonatal death for infants of mothers with cervical cancer was found. PMID- 11408875 TI - The presence of hepatitis B surface antigen and deoxyribonucleic acid in amniotic fluid and cord blood. AB - OBJECTIVE: It is uncertain whether neonatal infection with hepatitis B, despite treatment after delivery with immunoglobulin and vaccine, is the result of prior in utero transmission of the virus or treatment failure. Furthermore, the potential risk of hepatitis B transmission from the mother to the fetus at the time a genetic amniocentesis is performed is also a concern. In an attempt to better elucidate these controversies, amniotic fluid and cord blood specimens obtained from pregnant women positive for hepatitis B surface antigen were analyzed for the presence of hepatitis B surface antigen and hepatitis B deoxyribonucleic acid. STUDY DESIGN: This study was a prospective longitudinal analysis that identified hepatitis B surface antigen-positive patients who presented for amniocentesis. Cord blood was obtained from these patients at the time of delivery. Cord blood was also obtained from a group of hepatitis B surface antigen-positive patients for whom no amniocentesis was performed. All samples were analyzed for the presence of hepatitis B surface antigen and hepatitis B deoxyribonucleic acid. RESULTS: A total of 121 hepatitis B surface antigen-positive pregnant women were identified. In the 72 pregnancies in which amniocentesis was not performed, 18% of the cord blood samples were positive for hepatitis B surface antigen and 4% were positive for hepatitis B deoxyribonucleic acid. Of 47 amniocentesis fluid samples, 32% were positive for hepatitis B surface antigen but all were negative for hepatitis B virus deoxyribonucleic acid. Of 30 cord blood samples from patients who underwent an amniocentesis, 27% were positive for hepatitis B surface antigen, but all were negative for hepatitis B virus deoxyribonucleic acid. CONCLUSIONS: This study found that hepatitis B viral deoxyribonucleic acid is rarely present in cord blood and was not identified in amniotic fluid obtained by amniocentesis. This finding suggests that in utero transmission of the virus is rare prior to the onset of labor. These data further confirm the reports in the current literature that the risk of hepatitis B transmission to the fetus during amniocentesis is low. Because hepatitis B surface antigen can exist as an isolated entity devoid of nuclear material, in some cases this protein may be able to traverse the placental and amniotic membrane barrier in a manner similar to other proteins, such as alpha fetoprotein. Recommendations for genetic amniocentesis in women positive for hepatitis B surface antigen are discussed. PMID- 11408876 TI - Variation in elective primary cesarean delivery by patient and hospital factors. AB - OBJECTIVE: Our purpose was to describe variation in elective primary cesarean rates by nonclinical factors. STUDY DESIGN: With use of California discharge data and American Hospital Association data for 1995, patients were classified into 13 mutually exclusive categories for elective primary cesarean delivery. With use of recursive partitioning algorithms, women in each category were then studied to determine whether nonclinical factors were associated with elective primary cesarean delivery. RESULTS: A total of 463,196 women were delivered at 288 hospitals, and the elective primary cesarean delivery rate was 4.25% (19,664/463,196). Risk for elective primary cesarean delivery varied by clinical condition. The most discriminant risk factors were hospital type (malpresentation, multiple gestation, macrosomia, other hypertension), maternal age (antepartum bleeding, uterine scar, soft tissue disorder, preterm, unspecified), and teaching status (herpes, severe hypertension, unengaged head). CONCLUSION: This article presents methods that use administrative data to isolate and monitor the impact of nonclinical factors on the use of elective primary cesarean. PMID- 11408877 TI - Using active management of labor and vaginal birth after previous cesarean delivery to lower cesarean delivery rates: aA 10-year experience. AB - OBJECTIVE: Our objective was to analyze the statistics on cesarean delivery rates and the factors that have led to a reduction in these rates. STUDY DESIGN: A retrospective analysis was done of delivery statistics from a 10-year period, January 1, 1989, to December 31, 1998. We investigated the changes made in the methods of delivery during the study period. The data were divided into 1-year periods and analyzed by chi(2) tables. RESULTS: The overall cesarean delivery rate decreased from 16.59% to 10.92%; the primary cesarean delivery rate decreased from 9.22% to 7.11% and the repeated cesarean delivery rate from 7.37% to 3.81%. All these decreases were statistically significant. An increase in the rate of active management of labor by increasing oxytocin use and encouraging a trial of labor after previous cesarean delivery was also statistically significant. No changes in the outcome were observed in terms of neonatal morbidity and mortality rates. CONCLUSION: We found that our working plan for management of labor and delivery yielded and maintained a successful decline in the cesarean delivery rates without any negative effect on neonatal or maternal mortality rates. This low rate was maintained for a 10-year period. PMID- 11408878 TI - A retrospective study of the effect of postoperative indomethacin rectal suppositories on the need for narcotic analgesia in patients who had a cesarean delivery while they were under regional anesthesia. AB - OBJECTIVE: This study was undertaken to assess the efficacy of rectal indomethacin as an analgesic after cesarean delivery, comparing narcotic usage in patients receiving indomethacin with patients who received only standard narcotic analgesics. STUDY DESIGN: A retrospective study of patient records consisting of 297 women who underwent cesarean delivery while they were under regional anesthesia in a medium-sized community hospital from February 1996 through December 1999. Narcotic usage was determined as morphine equivalents whereby 10 mg morphine sulfate equals 30 mg oxycodone equals 50 mg hydrocodone equals 75 mg meperidine. Indomethacin was administered as 50-mg rectal suppositories. Statistical analysis was done by Pearson correlation and independent Student t test analysis. RESULTS: Narcotic usage was reduced by an average of 16 morphine equivalents (28%) in the indomethacin group. Pearson correlation revealed correlation at the.01 level; independent Student t test analysis revealed a P value of.001. Thus the mean amount of narcotic analgesic administered was significantly less in patients treated with indomethacin rectal suppositories. CONCLUSION: In this uncontrolled trial the use of indomethacin rectal suppositories resulted in a significant reduction in narcotic use in the postcesarean hospital recovery period as measured in morphine equivalents. PMID- 11408879 TI - The association of abnormal alpha-fetoprotein and adverse pregnancy outcome: does increased fetal surveillance affect pregnancy outcome? AB - OBJECTIVE: Our purpose was to investigate whether adverse outcomes associated with elevated maternal serum alpha-fetoprotein levels may be prevented by intensive antenatal monitoring. STUDY DESIGN: Records of patients with elevated maternal serum alpha-fetoprotein values of > or =2.0 multiples of the median between 1995 and 1999 were reviewed. Pregnancy histories were analyzed to determine whether intensive antenatal monitoring (twice-weekly nonstress tests and determinations of the amniotic fluid index) would have detected the adverse outcomes when routine obstetric care would have missed them. Women with elevations explained by multiple gestations, structural abnormalities, or a fetal death were excluded. RESULTS: The study enrolled 136 patients. Twenty-three patients were excluded because of multiple gestations, structural or chromosomal abnormalities, or fetal death or for lack of available follow-up. Seventy-eight patients had no perinatal complications, but 12 of these patients underwent heightened surveillance. One of these patients was subjected to an induction of labor. Thirty-five pregnancies had complications (21 with preterm labor, 7 with pregnancy-induced hypertension, 6 with growth restriction or oligohydramnios, 1 with abruptio placentae, and 1 with vasa previa). Of these 35 pregnancies, 22 were followed up with routine obstetric care and 13 with heightened surveillance. Heightened surveillance did not achieve earlier or improved detection in this group. These results suggest that routine pregnancy management is an adequate strategy for providing care to pregnant patients with unexplained elevated maternal serum alpha-fetoprotein levels. Adverse outcomes were detected with routine pregnancy management or were undetectable even with intensive management. CONCLUSION: Increased risks of pregnancy-induced hypertension, preterm delivery, intrauterine growth restriction, intrauterine fetal death, oligohydramnios, and abruptio placentae are associated with elevated maternal serum alpha-fetoprotein levels. However, in our study, routine pregnancy management was an acceptable method of detecting these adverse outcomes when they were detectable. PMID- 11408880 TI - Uterine artery embolization for the treatment of uterine fibroids: an outpatient procedure. AB - OBJECTIVE: Our objectives were to establish an outpatient program for uterine artery embolization of fibroids and to monitor the following: percentage of patients who required immediate hospitalization or admission within 2 weeks, outcomes in terms of the degree of ultrasound regression of the fibroids, patient satisfaction, reduction of pressure symptoms, and reduction of bleeding. STUDY DESIGN: Patients were screened by a gynecologist with the use of a designed care algorithm; they then underwent uterine embolization, performed by an interventional radiologist. Patients were evaluated at 6 weeks and 6 months after the procedure, and ultrasound studies were performed both before and at 2 to 6 months after the procedure. RESULTS: Of 35 patients, 29 (83%) went home on the day of the procedure, whereas 6 were observed overnight. Three (9%) patients required admission within 1 week. Of 26 patients, 24 (92%) were satisfied with the reduction of bleeding, and 14 of 18 (78%) were satisfied with the reduction in pressure symptoms. The mean decrease in uterine volume was 36%, and the mean decrease in the size of the dominant fibroid was 49%. CONCLUSIONS: Uterine artery embolization for the treatment of uterine fibroids might be done on an outpatient basis with a low rate of same-day admissions and delayed admissions. Patient satisfaction was high, and uterine artery embolization might become an accepted option for the treatment of uterine fibroids. PMID- 11408881 TI - Comparison of pain and time of procedures with two first-trimester abortion techniques performed by residents and faculty. AB - OBJECTIVES: We compared pain perception and procedure time in abortions performed by residents and faculty using a manual vacuum aspirator and electric vacuum curettage devices. STUDY DESIGN: We conducted a randomized trial of 114 women undergoing first-trimester abortions. Patients assessed the level of pain with visual analog scales. RESULTS: The mean procedure times were 5.7 and 6.9 minutes, respectively, with electric vacuum curettage and manual vacuum aspirator. Faculty took less time than residents to perform both procedures. Patients reported a higher pain level with cervical dilatation before resident electric vacuum curettage procedures. Patients undergoing electric vacuum curettage thought that the procedure noise increased their pain. CONCLUSIONS: First-trimester abortion procedures can be performed more quickly by experienced surgeons. The procedure time for the manual vacuum aspirator is greater than that for the electric vacuum curettage. Patient pain perception with aspiration by these two techniques is not different. The level of pain after aspiration did not vary significantly in patients who had abortions performed by residents or faculty. PMID- 11408882 TI - A comparison of echocardiography and pulmonary artery catheterization for evaluation of pulmonary artery pressures in pregnant patients with suspected pulmonary hypertension. AB - OBJECTIVE: This study was undertaken to compare the accuracy of echocardiography versus pulmonary artery catheterization to estimate pulmonary artery pressures in pregnant women with suspected pulmonary hypertension. STUDY DESIGN: A retrospective chart review was performed between January 1990 and February 2000 for all pregnant patients with cardiac disease. Patients with pulmonary artery pressure values estimated by cardiac catheterization and echocardiography during pregnancy were included. Pulmonary hypertension is defined as pulmonary artery systolic pressure >30 mm Hg. RESULTS: Twenty-seven patients were included in the study. There was a significant overestimation of the mean pulmonary artery pressure with echocardiography compared with catheterization (55.4 vs 51.1 mm Hg; P <.005). Of the 20 patients, pulmonary artery pressure was significantly greater when estimated by echocardiography than when measured by catheterization (59.6 vs 54.8 mm Hg; P <.004). Thirty-two percent (8/25) of the patients had pulmonary hypertension when estimated by echocardiography but had normal pulmonary artery pressures on subsequent catheterization. CONCLUSION: Echocardiography significantly overestimated pulmonary artery pressures compared with catheterization in pregnant patients with suspected pulmonary hypertension. Patients with structural cardiac defects appear to have a significantly greater difference in pulmonary artery pressures. Thirty-two percent of pregnant patients with normal pulmonary artery pressures may be misclassified as having pulmonary artery hypertension when measured by echocardiography alone. PMID- 11408883 TI - The second stage of labor and stress urinary incontinence. AB - OBJECTIVE: This study was undertaken to investigate the potential association between prolonged second stage of labor and stress urinary incontinence. STUDY DESIGN: A retrospective, population-based study was performed. A random, case controlled sample of 85 cases and 88 controls was identified by means of a standard computerized patient database. Subjects were identified by International Classification of Diseases, Ninth Revision codes, and medical records were reviewed. The median follow-up time from delivery was 7.8 years for cases and 7.2 years for controls. Multiple logistic regression was performed to test for an association between stress urinary incontinence and variables of interest. RESULTS: The data suggest that for all women who labored the length of the second stage of labor for the first delivery was not associated with stress urinary incontinence (odds ratio, 1.07; P =.42; 95% confidence interval, 0.9-1.3). However, forceps delivery was associated with a significant increase in stress urinary incontinence risk (odds ratio, 10.4; P =.04; 95% confidence interval, 1.2 93.4). CONCLUSION: Length of second stage of labor was not associated with stress urinary incontinence. However, the odds of having a later diagnosis of stress urinary incontinence was 10 times higher for women who underwent forceps delivery. PMID- 11408884 TI - Maternal and neonatal outcomes after uterine rupture in labor. AB - OBJECTIVE: There is significant controversy about the risks related to attempted vaginal birth after cesarean and the implications for informed consent of the patient. Recent data suggest that women who deliver in hospitals with high attempted vaginal birth after cesarean rates are more likely to experience successful vaginal birth after cesarean, as well as uterine ruptures. We conducted a study to evaluate maternal and neonatal morbidity and mortality after uterine rupture at a tertiary care center. STUDY DESIGN: We performed a retrospective chart review of cases of uterine rupture from 1976 to 1998. All women who had a history of uterine rupture were identified with International Classification of Diseases, Ninth Revision, identifiers with hospital discharge data cross-referenced with a separate obstetric database. We abstracted demographic information, fetal heart rate patterns, maternal pain and bleeding patterns, umbilical cord gas values, and Apgar scores from the medical record. Outcome variables were uterine rupture events and major and minor maternal and neonatal complications. RESULTS: During the study period there were 38,027 deliveries. The attempted vaginal birth after cesarean rate was 61.3%, of which 65.3% were successful. We identified 21 cases of uterine rupture or scar dehiscence. Seventeen women had prior cesarean deliveries (10 with primary low transverse cesarean delivery, 3 with unknown scars, 1 with classic cesarean delivery, 2 with two prior cesarean deliveries, and 1 with four prior cesarean deliveries). Of the 4 women who had no history of previous uterine surgery, one had a bicornuate uterus whereas the others had no factors increasing the risk for uterine rupture. We confirmed uterine rupture and scar dehiscence in 19 women. Specific details were not available for 2 patients. Uterine rupture or scar dehiscence was clinically suspected in 16 women with 3 cases identified at delivery or after delivery. Sixteen women had symptoms of increased abdominal pain, vaginal bleeding, or altered hemodynamic status. There were 2 patients who required hysterectomies and 3 women who received blood transfusions; there were no maternal deaths related to uterine rupture. The fetal heart rate pattern in 13 cases showed bradycardia and repetitive variable or late decelerations. Thirteen neonates had umbilical artery pH >7.0. Two cases of fetal or neonatal death occurred, one in a 23-week-old fetus whose mother had presented to an outlying hospital and the second in a 25-week-old fetus with Potter's syndrome. All live born infants were without evidence of neurologic abnormalities at the time of discharge. CONCLUSION: Our data confirm the relatively small risk of uterine rupture during vaginal birth after cesarean that has been demonstrated in previous studies. In an institution that has in-house obstetric, anesthesia, and surgical staff in which close monitoring of fetal and maternal well-being is available, uterine rupture does not result in major maternal morbidity and mortality or in neonatal mortality. PMID- 11408885 TI - Moral leadership and the issue of abortion. PMID- 11408886 TI - Moral leadership no excuse for invective. PMID- 11408888 TI - Amnioinfusion to prevent pulmonary hypoplasia after premature rupture of membranes. PMID- 11408890 TI - Effects of antenatal endotoxin on the lungs of preterm lambs. PMID- 11408892 TI - Safety and efficacy of laparoscopy for benign gynecologic conditions. PMID- 11408894 TI - Maternal effects of antenatal corticosteroid administration. PMID- 11408896 TI - Further concerns about the National Institute of Child Health and Human Development guidelines for interpretation of electronic fetal heart rate monitoring. PMID- 11408898 TI - Natural history of asymptomatic rotator cuff tears: a longitudinal analysis of asymptomatic tears detected sonographically. AB - The purpose of this study was to examine longitudinally the natural history of asymptomatic rotator cuff tears over a 5-year period and to assess the risk for development of symptoms and tear progression. Since 1985 through the present, bilateral sonograms were done on all patients. A review of consecutive sonograms done from 1989 to 1994 revealed 58 potential patients with unilateral symptoms who had contralateral asymptomatic rotator cuff tears. Of these 58 patients, 45 (22 men, 23 women) responded to a comprehensive questionnaire and 23 additionally returned for examination and repeat sonographic evaluation. The questionnaire was based on the American Shoulder and Elbow Surgeons score and included several outcome-based questions. A physical examination was performed in a standardized fashion along American Shoulder and Elbow Surgeons guidelines. Repeat high resolution sonograms were performed by a single experienced radiologist. Primary and repeat sonograms were then reassessed for tear size and location by two independent experienced radiologists blinded to the clinical data results. Twenty three (51%) of the previously asymptomatic patients became symptomatic over a mean of 2.8 years. The average Activities of Daily Living score for those remaining asymptomatic was 28.5 of 30 and for those becoming newly symptomatic, 22.9 of 30 (P <.5). The mean visual analog pain score (1 = no pain) for those remaining asymptomatic was 1.1 and for the newly symptomatic patients, 4.0. Of the 23 patients who returned for ultrasound, 9 were asymptomatic and 14 symptomatic. Only 2 of the 9 patients remaining asymptomatic had progression of their tears. Overall, 9 of 23 patients had tear progression. No patient had a decrease in the size of the tear. Our results demonstrate that symptoms can develop in patients with previously asymptomatic rotator cuff tears when seen in the context of a contralateral symptomatic tear. Development of symptoms was associated with a significant increase in pain and decrease in the ability to perform activities of daily living (P <.05). There appears to be a risk for tear size progression over time. PMID- 11408899 TI - Detection of acromioclavicular joint pathology in asymptomatic shoulders with magnetic resonance imaging. AB - The purpose of this study was to determine the prevalence of acromioclavicular joint (ACJ) arthritis with magnetic resonance imaging (MRI) evaluation in asymptomatic patients. Fifty shoulders in 42 patients were evaluated with the use of standard MRI techniques. There were 18 men and 24 women with an average age of 35 years (range, 19 to 72). ACJ arthritic changes were graded on a scale from 1 to 4 (none, mild, moderate, and severe), based on the amount of subacromial fat effacement, joint space narrowing, irregularity, capsular distension, and osteophyte formation. Forty-one (82%) of 50 shoulders had abnormalities consistent with arthritis on MRI. Patients were divided into two groups according to age: those older than 30 years and those 30 years old or younger. In the 30 and-under age group, 68% of the shoulders had arthritic changes, whereas in the over-30 age group, 93% had arthritic changes. Furthermore, in a comparison of the two groups, more advanced arthritic changes were found in the over-30 age group (P <.05). Clinical decisions to resect the ACJ should correlate the physical examination with the MRI findings because many patients may be clinically asymptomatic. PMID- 11408900 TI - Clinical evaluation of sodium hyaluronate for the treatment of patients with rotator cuff tear. AB - A randomized study of two types of conservative treatment of 78 shoulders in 78 patients with rotator cuff tears was performed. Twenty-five milligrams of sodium hyaluronate (SH) was injected into shoulders in one group (SH group) and 2 mg of dexamethasone was injected in the other group (steroid group). Injection of each drug was planned for once per week for 5 consecutive weeks. In the SH group, University of California at Los Angeles score before treatment was 13.6 +/- 2.6 points in 16 patients who did not require surgery (satisfied patients) and 12.8 +/- 3.5 points in 22 patients who required surgery (unsatisfied patients) (not statistically significant). In the steroid group, University of California at Los Angeles score before treatment was 11.9 +/- 3.6 points for 15 satisfied patients and 12.6 +/- 3.9 points for 25 unsatisfied patients (again, not statistically significant). In the SH group, the score at 4 weeks after treatment for satisfied patients increased to 27.6 +/- 3.1 points, whereas that for unsatisfied patients was 14.9 +/- 1.2 points (P <.0001). Similarly, in the steroid group, the score at 4 weeks after the treatment for satisfied patients increased to 26.5 +/- 2.0 points, whereas that for unsatisfied patients was 15.0 +/- 4.0 points (P <.0001). At 24 weeks after treatment, the score for satisfied patients was 26.2 +/- 3.1 points in the SH group and 25.3 +/- 2.5 points in the steroid group. The effective rate of the SH group was 39.5% and that of the steroid group was 35%. Therapeutic efficacy in the SH group was equivalent to that in the steroid group. In both groups, the rate of patients who engaged in manual labor was significantly higher in the group of unsatisfied patients than in that of satisfied patients. No adverse reaction to either treatment was observed. These results suggest that SH is an effective conservative treatment for patients with rotator cuff tears. PMID- 11408901 TI - Glenoid revision surgery after total shoulder arthroplasty. AB - Forty-eight shoulders that underwent glenoid component revision surgery were reviewed at a mean of 4.9 years (range, 2 to 12 years). The indications for surgery were glenoid component loosening in 29 shoulders, glenoid implant failure in 14 shoulders, and glenoid component malposition or wear leading to instability in 5 shoulders. Seventeen shoulders had associated instability. Thirty shoulders underwent implantation of a new glenoid component and 18 underwent removal of the component and bone grafting for bone deficiencies. There was significant pain relief, improvement in active elevation and external rotation, and satisfaction with revision glenoid surgery (P <.05). Patients without a glenoid component were significantly less satisfied with the procedure than those patients who underwent reimplantation of a glenoid component (P =.01). Satisfactory pain relief was achieved in 86% of patients with a new glenoid component and 66% of patients who underwent glenoid component removal. Seven shoulders with a new glenoid component (2 for glenoid loosening) and 5 who underwent removal without reimplantation (3 for painful glenoid arthritis) required re-revision surgery. Eleven of the 17 patients with instability were stable at the most recent follow-up. The data from this study suggest that at the time of revision glenoid surgery, patients who have placement of a glenoid component have a higher degree of satisfaction than those undergoing glenoid component removal. Patients who continue to have pain after bone grafting without placement of a component may be candidates for glenoid component placement after graft consolidation. PMID- 11408902 TI - Arthroscopic subacromial decompression. AB - In this study, we analyzed the results of two series of patients treated for impingement syndrome by undergoing arthroscopic subacromial decompression (ASD). Patients had not responded to nonoperative treatment. Group 1 included 112 consecutive patients (average age, 41 years) with 96 (77%) patients available for 2-year follow-up. Group 2 (28 patients, 29 shoulders; average age, 43 years; range, 22 to 72) had ASD and the subacromial space digitally palpated to determine if adequate decompression was performed. Twenty-two (85%) of 26 shoulders were available for follow-up. At follow-up, pain, function, range of motion, strength, impingement signs, and patient satisfaction were assessed. In group 1, according to the Neer criteria, 48% of the patients were graded as satisfactory and 52% unsatisfactory. Workers' Compensation patients had a satisfactory rate of 32%, whereas non-Workers' Compensation patients had a satisfactory rate of 59%. Twenty patients had open acromioplasty after ASD. Inadequate decompression was noted in 14 of 20 failed patients. In group 2, 86% of the patients were graded as satisfactory according to the Neer criteria, with 14% unsatisfactory, which included the 2 failures. The 2 (9%) of 22 shoulders that failed the ASD went on to further surgical treatment. Average follow-up was 56 months (range, 13 to 78 months). The average American Shoulder and Elbow Society score at follow-up was 90.4. No difference between Workers' Compensation cases and the other cases was seen (P <.7). Finger palpation can help to improve outcomes by allowing the surgeon to assess the adequacy of decompression. PMID- 11408903 TI - Axillary nerve injury after thermal capsular shrinkage of the shoulder. AB - The purpose of this study was to evaluate 4 cases in which axillary nerve injury was observed after radiofrequency capsular shrinkage of the shoulder. These cases were used to evaluate the clinical circumstances under which axillary nerve injury occurred, the time frame over which these injuries recovered, and the ultimate outcome observed in these patients. The case histories of each of these 4 patients were carefully evaluated. In 2 of the 4 cases, purely sensory findings were found, and in 2 cases both sensory and motor injuries occurred. In each case the inferior axillary recess was treated using the Oratech Interventions Tac-S probe. The authors conclude that injury to the axillary nerve during this procedure is possible and postulate that heat penetration through the capsule to the nerve is the most likely cause of injury. PMID- 11408904 TI - Rotator cuff repair in a district hospital setting: outcomes and analysis of prognostic factors. AB - In a prospective study of 58 complete rotator cuff tears, 54 patients underwent surgery by a single surgeon for chronic, symptomatic, full-thickness rotator cuff defects. An independent observer performed the evaluation. Fifty-two of 58 patients reported marked relief of pain and rated themselves as having mild or no deficit in their ability to perform daily activities. The study evaluated preoperative and intraoperative factors that influenced postoperative outcome. Eighty-three percent of patients had good or excellent results according to the modified Constant scoring system and 76% with the UCLA system. There was no pain or minimal pain in 90%, and 92% reported overall improvement after the surgery. It is concluded that in a patient with a full-thickness rotator cuff tear, pain and shoulder function can be improved by acromioplasty and cuff repair, but a slight decrease in the range of motion and muscle strength will probably remain. PMID- 11408905 TI - The effects of age, sex, and shoulder dominance on range of motion of the shoulder. AB - To determine the effects of age, sex, and arm dominance on shoulder range of motion, we measured active and passive forward elevation, abduction, internal and external rotation at 90 degrees of abduction, external rotation with the arm adducted, and extension bilaterally in 280 subjects ranging in age from 4 to 70 years. Linear regression analyses were performed for all motions except forward elevation. This motion, which showed a nonlinear pattern of decline with age, was evaluated with 3-way analysis of variance. Shoulder range of motion decreased with age for all measured motions with the exception of internal rotation, which increased with age. Female subjects had a significantly greater range of motion than male subjects for all motions measured. Dominant arms displayed significantly greater external rotation than nondominant, regardless of whether the arm was abducted or adducted at the time of measurement. However, nondominant shoulders demonstrated significantly greater internal rotation and extension than dominant. No significant differences were found between dominant and nondominant sides for forward elevation or abduction. PMID- 11408906 TI - Results of biceps tenotomy for treatment of pathology of the long head of the biceps brachii. AB - Historically, the surgical treatment of bicipital pathology has been a variety of tenodesis techniques. The purpose of this study is to report the results of simple biceps tenotomy for the treatment of bicipital pathology. Thirty shoulders in 30 consecutive patients who had a simple arthroscopic biceps tendon release were reviewed. Data was collected according to the method of the American Shoulder and Elbow Surgeons (ASES) shoulder evaluation form. Outcome was assessed with the rating system of the ASES. The mean ASES shoulder score was 81.8. There was a significant reduction in pain and improvement in function after the procedure. The complication rate was 13.3%. Bicipital pathology is a significant cause of morbidity around the shoulder. The results of this study demonstrate that functional outcome as measured by the ASES scoring system can be very good with an arthroscopic biceps tendon release for the treatment of biceps tendon pathology. PMID- 11408907 TI - Function of the long head of the biceps at the shoulder: electromyographic analysis. AB - The purpose of this study was to delineate, through electromyographic analysis, the function of the long head of the biceps at the shoulder. Ten shoulders were examined with dynamic electromyography. The long head of the biceps was instrumented with thin wire electrodes. The supraspinatus, infraspinatus, deltoid, brachialis, and brachioradialis were instrumented as controls. Because the biceps functions primarily as a forearm supinator and elbow flexor, a long arm brace was used to lock the elbow in extension with the forearm in neutral pronation/supination. Each motion was tested in a full arc at fast (170 degrees per second) and slow (36 degrees per second) speeds and repeated with and without a 5-pound weight attached to the distal end of the brace. No electrical activity was identified in the long head of the biceps muscle in response to isolated shoulder motion with the elbow and forearm position controlled. The data demonstrate that the long head of the biceps is not active in isolated shoulder motion when the elbow and forearm are controlled. Thus, any hypothesis on bicipital function at the shoulder must be based on either a passive role of the tendon or tension in association with elbow and forearm activity. PMID- 11408908 TI - Fractured rheumatoid elbow: treatment with Souter elbow arthroplasty--a clinical and radiologic midterm follow-up study. AB - We report the results in 26 patients who had 32 preoperative fractures treated with Souter elbow arthroplasty. All were rheumatoid patients with a mean disease duration of 29.7 years (range, 10 to 43). Six of the fractures were of the olecranon and 26 of the distal humerus. The time interval between fracture and arthroplasty was 9 months (mean; range, 0 to 48). Fragments were not excised, and osteosynthesis was performed. The follow-up was 2.6 years (mean; range, 0.5 to 8), when 20 of the fractures had united and 12 had not. K-wire fixation, either alone or in combination with cerclage or PDS suture, and bone grafting led to satisfactory results. Union was verified in 14 of 17 cases treated with this technique. There were no severe early complications. Six patients had late complications. In 3 cases, loosening of the humeral component was observed radiologically. One patient had a hematogenous deep infection 4 years after the operation, and 2 patients had avulsion rupture of the triceps tendon. Fracture in the badly destroyed elbow can be more reasonably treated with an arthroplasty than with an attempt of osteosynthesis before arthroplasty. If excision of the fragments is avoided, original, or near original, anatomy of the elbow joint can be better restored and acceptable outcome obtained with elbow arthroplasty. PMID- 11408909 TI - Role of collateral ligaments in the GSB-linked total elbow prosthesis. AB - The GSB III elbow prosthesis is a loose-hinged type of elbow implant. The introduction of such hinged elbow arthroplasty expanded the indications for elbow replacement to patients with more deficient bone and ligaments. The purpose of this study was to compare the kinematics and stability of the GSB III elbow prosthesis with that of the normal elbow and to investigate the role of the collateral ligaments in the kinematics and the stability of the GSB III total elbow prosthesis in an in vitro model. The results could show a semiconstrained kinematic pattern of the GSB III implant. The mean laxity for varus/valgus stress of the implant without collateral ligament support was significantly greater for all flexion angles when compared with intact elbows (mean, 12.7 degrees versus 5.4 degrees ) and with elbows treated with a standard implantation technique (mean, 9.5 degrees ). The release of the collateral ligaments increased the already observed varus shift after standard implantation of a GSB III elbow prosthesis. The laxity measured without collateral ligaments during loaded movement reached the maximum varus/valgus laxity of the GSB III prosthesis of 12 degrees degrees. The study confirms the role of the collateral ligaments in stabilizing the GSB III elbow prosthesis. Missing collateral ligaments may overload the implant-cement-bone interface and may be one factor contributing to early aseptic loosening of this device. PMID- 11408910 TI - The posterior subdeltoid approach: a modified access to the posterior glenohumeral joint. AB - A modified surgical approach to the posterior aspect of the glenohumeral joint and/or the dorsal glenoid is described. This access does not alter any muscle insertion or neuromuscular planes. After the skin incision is made, the inferior border of the spinal part of the deltoid is identified and the deltoid muscle is mobilized and retracted, thus offering an excellent approach to the interval between the infraspinatus and teres minor muscles. This interval is split parallel to the muscle fibers. This surgical approach was first established in 10 cadaverous shoulders and then performed in 12 patients with posterior shoulder pathology. In the cadaver study, the closest distance to the axillary nerve with this approach was 22 mm. In all 12 cases, the surgical procedure could be performed without any problems. PMID- 11408911 TI - Direct 3-dimensional measurement of scapular kinematics during dynamic movements in vivo. AB - The purpose of this study was to describe 3-dimensional scapular motion patterns during dynamic shoulder movements with the use of a direct technique. Direct measurement of active scapular motion was accomplished by insertion of 2 1.6-mm bone pins into the spine of the scapula in 8 healthy volunteers (5 men, 3 women). A small, 3-dimensional motion sensor was rigidly fixed to the scapular pins. Sensors were also attached to the thoracic spine (T3) with tape and to the humerus with a specially designed cuff. During active scapular plane elevation, the scapula upwardly rotated (mean [SD] = 50 degrees [4.8 degrees ]), tilted posteriorly around a medial-lateral axis (30 degrees [13.0 degrees ]), and externally rotated around a vertical axis (24 degrees [12.8 degrees ]). Lowering of the arm resulted in a reversal of these motions in a slightly different pattern. The mean ratio of glenohumeral to scapulothoracic motion was 1.7:1. Normal scapular motion consists of substantial rotations around 3 axes, not simply upward rotation. Understanding normal scapular motion may assist in the identification of abnormal motion associated with various shoulder disorders. PMID- 11408912 TI - Scapular and clavicular kinematics during humeral elevation: a study with cadavers. AB - A combination of kinematic testing and graphic reconstruction of cadaveric shoulders was used to characterize shoulder kinematics during a simulated passive clinical range-of-motion examination. Cadaveric shoulders were elevated in the coronal, scapular, and sagittal planes while the scapula, clavicle, and humerus were kinematically tracked. Graphic models of each shoulder were created from computed tomography data. The models were animated to display the experimental motions. Shoulder kinematics varied between elevation planes. The scapular and clavicular rotations were relatively small until the humerus reached approximately 90 degrees of elevation. Clavicular and scapular rotations that occurred at low humeral elevation angles for elevation in the coronal plane were significantly larger than for the other two planes. The glenohumeral to scapulothoracic ratio was approximately equal to 2 for the entire range of elevation for each elevation plane, but it was dramatically larger during early elevation than during late elevation. PMID- 11408913 TI - A novel, resorbable suture anchor: pullout strength from the human cadaver greater tuberosity. AB - The pullout strength of a collagen bone anchor that creates interference fixation as the result of radial swelling on hydration was compared with a Mitek rotator cuff anchor after insertion into the greater tuberosity of human cadaver humeri. Bones were fully hydrated at 37 degrees C. Stiffness, peak load, and the mode of failure were recorded. Real and apparent bone densities were measured. Peak load for the collagen anchor at 15 minutes (121.0N +/- 81.3N) was greater than at 2 minutes (60.5N +/- 38.5N) after insertion (P <.05). At between 5 and 60 minutes after insertion, peak loads for the Mitek and the collagen anchors did not differ. After 30 minutes from insertion, the mode of failure of the collagen anchor changed from pullout with minor body damage to pullout with major body damage. Peak load at pullout correlated with bone density for the Mitek (P <.05, r = 0.516) but for the collagen bone anchor appeared unaffected by bone density. PMID- 11408914 TI - Chronic plasma cell osteomyelitis of the humerus associated with Shigella and Flavobacterium. PMID- 11408915 TI - Osteochondritis dissecans developing in the trochlea humeri: a case report. PMID- 11408916 TI - Targeting new anticancer drugs within signalling pathways regulated by the Ras GTPase superfamily (Review). AB - A dynamic equilibrium or is responsible for the proper function of a living organism. Physiological events regulating proliferation, apoptosis, differentiation, and cell arrest, modulates the correct homeostasis and functionality of all tissues. Cancer is a consequence of a disorder in these sequential events, which results in the alteration of the ratio between cell death, cell differentiation and cell proliferation that ultimately leads to an increase in the number of dysregulated cells. Most of the processes which control the are regulated by signalling pathways, whose components are currently being explored as potential targets for the design of antitumoral drugs. Many in vivo studies have shown that Ras and Rho proteins are key modulators of mitogenic signalling, and are involved in the carcinogenesis of several human tumors. The development of recent drugs that elicit antitumoral activity by blocking some of the Ras and/or Rho effects, is discussed in this review. PMID- 11408917 TI - Activation of caspase-3 in renal cell carcinoma cells by anthracyclines or 5 fluorouracil. AB - The caspase family of proteases is speculated to have a crucial role in apoptosis. The effect of treatment with adriamycin (ADR), cisplatin (CDDP), 5 fluorouracil (5-FU), vinblastine (VLB), IFN-alpha, or IFN-gamma on the activation of caspase-3, -6, -8, and -9 in renal cell carcinoma (RCC) cells was investigated, to clarify the mechanisms of chemo- and immunotherapeutic agent mediated apoptosis. Caspase activity was determined by a quantitative colorimetric assay. Apoptosis was monitored by acridine-orange staining assay. Treatment of ACHN cells with CDDP, VLB, IFN-alpha, or IFN-gamma did not activate caspase-3, but its activity was increased 7.2-fold (p = 0.0001) with ADR and 2.8 fold (p = 0.0385) with 5-FU in comparison with control. Furthermore, when the ADR treatment time was shortened from 24 to 8 or 2 h, the same caspase-3 activation occurred. Activation of caspase-3 was also observed in six freshly isolated human RCC cells after the treatment with ADR. Of the six freshly derived RCC cells treated with 5-FU, caspase-3 activity was increased 3.1-fold (p = 0.0051) and 2.4 fold (p = 0.0346) in two of them, respectively. Epirubicin and pirarubicin, compounds closely related to ADR, also respectively enhanced 4.2-fold (p = 0.0052) and 2.8-fold (p = 0.0147) caspase-3 activity in ACHN cells. The activation of caspase-3 observed with a colorimetric assay was confirmed with immunocytochemical analysis using the anti-active caspase-3 mAb, which specifically recognizes the active form of caspase-3. Furthermore, both active caspase-3 and apoptosis triggered by either ADR or 5-FU were inhibited significantly by the general caspase inhibitor Z-VAD-FMK, or a specific caspase-3 inhibitor DMQD-CHO. These findings provide a mechanistic explanation for anthracyclines and 5-FU induced-apoptosis. PMID- 11408918 TI - Human telomerase reverse transcriptase antisense treatment downregulates the viability of prostate cancer cells in vitro. AB - Telomerase, a ribonucleoprotein complex is activated in the vast majority of human malignancies, including prostate cancer. Its inhibition is a putative way to affect cancer proliferation and might be used in the therapy of tumors. We analysed the influence of antisense phosphorothioate oligonucleotides (PTO) against the reverse transcriptase subunit of telomerase on prostate cancer cell viability, telomerase activity and telomere length. DU145 prostate cancer cells were cultivated in PTO containing medium. The PTO-incorporation was confirmed by confocal laser scanning microscopy. Cell viability was measured by a WST-1 tetrazolium assay. After 15 days of antisense PTO treatment, a significant inhibition of cell viability occurred. Telomerase activity was determined by a telomeric repeat amplification protocol (TRAP) assay and telomere length by Southern blot analysis. Since the long-term telomerase antisense treatment reduces the viability of prostate cancer cells significantly, this antisense approach could be a new therapeutic strategy to treat patients with advanced prostate cancer. PMID- 11408919 TI - Steroid receptors and hormones in relation to cell proliferation and apoptosis in poorly differentiated epithelial ovarian tumors. AB - The purpose of this study was to further investigate the role of estrogen but especially progesterone on epithelial ovarian tumor development since previous studies have suggested a relationship between serum progesterone, progesterone receptor expression and prognosis. Serum progesterone concentration, the immunohistochemical expression of estrogen receptor alpha (ER), progesterone receptor A/B (PR), Ki-67, Bcl-2, p53, apoptosis and morphology were determined in 33 patients, all with poorly differentiated surface epithelial ovarian tumors of different types. ER was expressed in 79% and PR in 33% of the tumors. This group of aggressive tumors was highly proliferative as indicated by Ki-67 index (mean 38.9%), and in some cases proliferation appeared to be mainly located to areas with a high ER density. The majority of cases (76%), both receptor-positive and negative, overexpressed p53. High ER expression was related to a lower apoptotic activity as compared with tumors with a low expression of the ER (p = 0.008). Serum progesterone in itself did not show any clear relationship to steroid receptor status, expression of Ki-67, p53, Bcl-2 or signs of apoptosis. Survival in this small but homogeneous group of advanced epithelial ovarian cancers, showed an improved survival rate in patients with high serum progesterone, especially in combination with expression of progesterone receptors (p = 0.04). In conclusion, estrogen and progesterone receptors in parallel with deranged p53 and Ki-67 were expressed to a great extent. The finding of a lower apoptotic activity in tumors with a high expression of ER and an indication of increased proliferation in areas with high ER density gives a rationale for antiestrogen therapy even in poorly differentiated epithelial ovarian cancers. Improved survival is related to serum progesterone, especially in combination with PR expression. PMID- 11408920 TI - Expression of second class of KIP protein p57KIP2 in human colorectal carcinoma. AB - p57KIP2, the second class of KIP family protein, is one of the negative regulators of the cell cycle. To elucidate the role of p57KIP2 in colorectal normal mucosa and cancer, we examined the expression of p57KIP2 protein in 110 pairs of colorectal non-tumor and cancer tissues. Immunohistochemical analysis showed that p57KIP2 was weakly detected in the normal colonic epithelium and lymph follicles. A unique expression pattern of p57KIP2 was exclusively noted in the elastic fibers within the walls of relatively large blood vessels (diameter > 100 microm). In cancer tissues, p57KIP2 protein was localized mainly in nuclei. Using the mean percentage of nuclear p57KIP2 expression (25%) as the cut-off value, we divided our cases into those with high expression (n = 44, 40%) and low expression (n = 66, 60%) of p57KIP2 among 110 colorectal cancer cases tested. The clinical and pathological survey showed a significant correlation between low expression of p57KIP2 and large tumor size (p < 0.05) or the presence of tumors in females (p < 0.01). Survival analysis showed that p57KIP2 expression did not influence prognosis. RT-PCR analysis was also performed using RNA extracts from 6 colorectal cancer tissues. When the levels of p57KIP2 mRNAs were compared with expression of p57KIP2 protein, a clear correlation was found, suggesting that expression of the p57KIP2 protein may be regulated at the transcription level. The present study revealed p57KIP2 expression in colorectal cancer and suggests that p57KIP2 may not play a central role in the progression of colorectal cancer. PMID- 11408921 TI - Loss of p27 expression predicts poor prognosis in patients with Dukes' B stage or proximal colorectal cancer. AB - p27 is a cyclin-dependent kinase inhibitor which regulates progression of cells from G1 into S phase in a cell cycle. Loss of the negative regulator may contribute to oncogenesis and tumor progression. The aim of this study was to examine p27 expression in normal mucosa, primary and metastatic tumors from patients with colorectal adenocarcinomas and to analyze association of p27 with patient survival and clinicopathological variables. p27 expression was estimated by immunohistochemistry in 178 primary colorectal cancers, 34 lymph node metastases and 48 normal mucosa samples from patients with colorectal adenocarcinoma. Associations of p27 with patient survival, clinicopathological characteristics and expression of p53, p73 and DCC were analyzed. Loss of p27 was found in 51% of primary tumors, 68% of metastases and 56% of normal samples. The intensity of p27 staining was similar in the matched primary tumor, metastasis and normal mucosa. In patients with Dukes' B or with proximal tumors, the loss of p27 predicted poorer prognosis (p = 0.03 and p = 0.05, respectively). However, there were no significant differences in the patients with other individual Dukes' stage or distal tumors. No relationships were found between p27 and patients' gender, age, tumor location, growth pattern and expression of p53, p73 and DCC (p > 0.05). The data suggest that loss of p27 was associated with poor prognosis in patients with Dukes' B tumor or those with proximal tumor. p27 might be a useful marker to identify the more progressive tumors in these groups. PMID- 11408922 TI - Prognostic predictive value of 18F-fluorodeoxyglucose positron emission tomography for patients with pancreatic cancer. AB - 18Fluorine-fluorodeoxyglucose positron emission tomography (18F-FDG-PET) is a unique imaging diagnostic tool to evaluate glucose metabolism and hexokinase activity which may reflect the aggressiveness of a tumor. Thirty-seven patients with primary pancreatic cancer were evaluated with 18F-FDG-PET. Thirteen patients underwent resection for the pancreatic cancer and 24 patients had unresectable tumors. The standardized uptake values (SUV) of 18F-FDG in the primary tumors were calculated. No correlations were found between the SUV in the tumors and the metastatic status to the peritoneal/liver, TNM factors/stage, or resectability. The patients were divided into 2 groups with high and low SUV, with the cut-off value being 3.0 (median SUV value of 37 cases). No differences in the probability of survival were observed between the 2 groups in the patients with resectable tumors. However, in the patients with unresectable tumors, those in the high SUV group had a significantly shorter prognosis than those in the low SUV group. Moreover, a multivariate analysis of survival indicated that SUV is an independent prognostic factor for patients with unresectable pancreatic cancer. PMID- 11408923 TI - Expression of galectin-3-reactive ligands in squamous cancer and normal epithelial cells as a marker of differentiation. AB - The definition of biological markers for oropharynx and larynx cancer is essential to predict their clinical behavior. Since cellular glycans play an important role in biological information transfer, we have employed an endogenous lectin, galectin-3, to examine in primary squamous carcinomas, lymph node metastases, and physiological squamous epithelia whether glycans recognized by this lectin are altered in relation to the state of differentiation. The expression of galectin-3 was concomitantly evaluated by immunohistochemistry using the A1D6 monoclonal antibody. In addition, other antibodies were used for the detection of cytokeratins and desmosomal proteins (desmoplakin-1 and desmoglein). The results show the expression of galectin-3-reactive ligands in moderately/highly differentiated carcinomas only in areas exhibiting a high level of keratinization. Except for one patient out of 14, metastatic cells in lymph nodes expressed no accessible binding sites for galectin-3. No galectin-3 reactivity was detected in the basal cell layer of all studied normal epithelia (which contains the proliferating cells). The suprabasal layers were positive in epidermis and epithelium of tongue and cornea and negative in epithelium of palatine tonsil. The tumor cells expressed galectin-3 with an intensity positively correlated with tumor differentiation. The position of galectin-3 reactive sites colocalized with the two tested desmosomal proteins. However, presence of these proteins was also detected in areas of tumor and suprabasal layers of tonsil epithelium where no binding reactivity for galectin-3 was found. The present study showed that expression of galectin-3-reactive glycoligands is differentiation-dependent in normal as well as malignant squamous cells. Colocalization of galectin-3-reactive sites with desmosomal proteins (desmoplakin 1 and desmoglein) suggests an association of the galectin-3 ligand(s) with the cell surface, pointing to a potential participation of galectin-3 in mediation of intercellular contacts in these tumor types. PMID- 11408924 TI - Accumulation of extracellular matrix in the liver induces high metastatic potential of hepatocellular carcinoma to the lung. AB - The liver undergoes pathogenic changes such as hepatitis, fibrosis and cirrhosis under continuous stimulation by hepatitis virus or alcohol intake, leading to the development of hepatocellular carcinoma. The metastatic potential of HCC can be positively or negatively regulated by pathogenic alterations of liver. We investigated whether the metastatic abilities of HCC after orthotopic implantation can be influenced in the fibrotic liver by continuous injection of carbon-tetrachloride (CCl4) for seven weeks. The incidence of lung metastasis after orthotopic implantation of murine HCC (CBO140C12) fragments into CCl4 treated livers was higher than into normal livers. The amount of mRNA for MMP-2 increased in the CCl4-treated livers as compared with normal livers, and CBO140C12 cells constitutively expressed mRNA for MT1-MMP in early amplification cycles by RT-PCR. In addition, we found that the culture of CBO140C12 cells on the substrates pre-coated with ECM components increased the expression of MMP-2 mRNA. Thus, enhanced incidence of lung metastasis in the fibrotic liver might be partly due to: i) over-expression of MMP-2 in the fibrotic liver in cooperation with MT1-MMP on the CBO140C12 cell surface, ii) over-expression of MMP-2 in CBO140C12 cells, possibly mediated by the interaction of tumor cells (surface integrins) with accumulated ECM in the fibrotic liver. This is the first report showing that increase of MMP-2 in the fibrotic liver can influence the metastatic potential of HCC cells. PMID- 11408925 TI - Altered expression of TGFB receptors and mitogenic effects of TGFB in pancreatic carcinomas. AB - Alteration of the transforming growth factor beta (TGFB) signalling pathway is important in pancreatic carcinogenesis, as shown by the frequent inactivation of the downstream target SMAD4. We recently analysed a series of pancreatic carcinoma cell lines with respect to alterations of five SMAD genes involved in TGFB signalling, and showed that SMAD4 was structurally rearranged in 42% of these. This pathway may, however, also be affected by alterations of genes whose products regulate the activation of TGFB as well as of TGFB receptor genes. We therefore studied the expression of UPA, UPAR, IGF2R, ALK5 (TGFBR1), TGFBR2, TGFBR3, ENG, ALK1, TGFB1, TGFB2, and TGFB3 in a series of 14 pancreatic carcinoma cell lines. We also analysed ALK5 and TGFBR2 for mutations, cell surface localisation of TGFBR2 and ENG, and TGFB1 response. No mutations of ALK5 or TGFBR2 were found. However, 4 cell lines were methylated within the ALK5 promoter region. ALK5 expression was strongly reduced in 9 cases, whereas TGFBR2 expression was increased in 12 of the cell lines. The TGFB signalling associated receptors ENG and ALK1 were co-expressed in 4 of the cell lines. There was no evidence for disruption of the UPAR-IGF2R TGFB activating pathway. The response to TGFB1 was analysed in 12 cell lines, and 6 of these (50%) showed increased proliferation. The cell lines stimulated by TGFB showed frequent mutations of SMAD4, KRAS2, and TP53, as well as frequent absence of CDKN2B expression. These results suggest that the ALK5-SMAD4 part of the TGFB signalling pathway is a major target for inactivation in pancreatic carcinomas, that the expression of TGFBR2, TGFBR3, and receptors involved in TGFB activation are maintained, and that alterations of components of the TGFB signalling pathway may be accompanied by a positive effect of TGFB on cell growth. PMID- 11408926 TI - Trans-resveratrol, a grapevine-derived polyphenol, blocks hepatocyte growth factor-induced invasion of hepatocellular carcinoma cells. AB - We have shown that liver myofibroblasts stimulate in vitro invasion of hepatocellular carcinoma cell lines through a hepatocyte growth factor/urokinase dependent mechanism. Resveratrol, a grapevine-derived polyphenol, has been shown to inhibit cellular events associated with tumor initiation, promotion and progression. The aim of this study was to evaluate the effects of trans resveratrol on invasion of the human hepatoma cell line HepG2. Cell invasion was assessed using a Boyden chamber assay. Activation of the HGF signal transduction pathways was evaluated by Western blot with phospho-specific antibodies. Urokinase expression was measured by RT-PCR and zymography. Trans-resveratrol decreased hepatocyte growth factor-induced cell scattering and invasion. It also decreased cell proliferation without evidence for cytotoxicity or apoptosis. Trans-resveratrol did not decrease the level of the hepatocyte growth factor receptor c-met and did not impede the hepatocyte growth factor-induced increase in c-met precursor synthesis. Moreover, trans-resveratrol did not decrease hepatocyte growth factor-induced c-met autophosphorylation, or Akt-1 or extracellular-regulated kinases-1 and -2 activation. Finally, it did not decrease urokinase expression and did not block the catalytic activity of urokinase. In conclusion, our results demonstrate that trans-resveratrol decreases hepatocyte growth factor-induced HepG2 cell invasion by an as yet unidentified post-receptor mechanism. PMID- 11408927 TI - Examination of the oncogenic potential of H19 gene in HeLa x normal human fibroblast hybrid cells. AB - Fusion of tumorigenic HeLa cells with human skin fibroblasts results in chromosomally stable non-tumorigenic hybrids. The studies of rare spontaneous tumorigenic segregants from the non-tumorigenic hybrid have implicated the loss of one copy of human fibroblast chromosome 11 with concomitant re-expression of tumorigenicity. In a previous study of differential display screening, we reported the tumorigenic-specific expression of H19 gene as a possible candidate for elicitation of tumorigenic phenotype. In this study, we examined the expression of H19 gene using gamma-ray-induced tumorigenic mutants (GIMs) and non tumorigenic irradiated control cells (CONs) from the non-tumorigenic hybrids. H19 expression was recognized in all the GIMs with various expression levels, whereas no CONs expressed the H19 gene. To examine the tumorigenic potentials of H19 gene directly, we introduced an H19 gene expression vector into the non-tumorigenic hybrids and assayed the tumorigenicity of the transfectants by s.c. injection into athymic nude mice. However, no transfectants with stable H19 gene expression induced in vivo tumor growth. These results suggest that expression of the H19 gene may be necessary but is not sufficient to confer the tumorigenic phenotype in HeLa x fibroblast hybrids. PMID- 11408928 TI - Immortalization of human esophageal keratinocytes by E6 and E7 of human papillomavirus type 16. AB - Transduction of human papillomavirus type 16 (HPV16) E6/E7 into primary culture of human esophageal keratinocytes using a recombinant adenovirus prolonged the life-span, while untreated cells senesced within 14-16 population doublings (PDLs). Up-regulation of telomerase activity and acquisition of serum-resistant growth were observed in the esophageal keratinocytes with extended life-span between 50 and 100 PDLs, and drastically increased after 100 PDLs. A keratinocyte sample with a polymorphism of Pro/Pro at codon 72 of p53 showed resistance to HPV16 E6/E7-induced life-span-extension and immortalization, in contrast to others with p53 polymorphisms of Arg/Arg or Arg/Pro, which did not. The high efficiency of E6/E7-induction by adenovirus vector also revealed the M1 and M2 stages of keratinocyte immortalization first described in this report. PMID- 11408929 TI - Molecular cloning and characterization of human Frizzled-5 gene on chromosome 2q33.3-q34 region. AB - Hfz5 is a potent cancer associated gene, encoding WNT receptor with the potential to activate beta-catenin - TCF signaling pathway. Here, human Frizzled-5 (FZD5) gene and cDNAs were cloned and characterized. FZD5 was almost identical to Hfz5, except for six amino-acid substitutions at codon 88, 262, 263, 345, 357, and 402. HF5S1 probe (nucleotide position 2036-2535 of FZD5 cDNA) hybridized to 7.5- and 3.5-kb FZD5 mRNAs, and HF5S2 probe (nucleotide position 5572-6194 of FZD5 cDNA) hybridized only to 7.5-kb FZD5 mRNA. FZD5 cDNA was polyadenylated at the nucleotide position 6534, while several FZD5 ESTs were polyadenylated at the nucleotide position 2561. The 7.5- and 3.5-kb FZD5 mRNAs were transcribed probably due to alternative splicing. FZD5 was highly expressed in fetal liver and adult pancreas, and moderately expressed in fetal lung, kidney and adult liver. Among human cancer cell lines, FZD5 was highly expressed in K-562 cells derived from chronic myelogenous leukemia. FZD5 gene, consisting of two exons, was mapped to human chromosome 2q33.3-q34 region, near the FZD7 gene and the FRA2I fragile site. These results suggest that FZD5 up-regulation might play key roles in chronic myelogenous leukemia through activation of the WNT - beta catenin - TCF signaling pathway. PMID- 11408930 TI - Up-regulation of Frizzled-7 (FZD7) in human gastric cancer. AB - Human Frizzled-7 (FZD7) and human FzE3, showing 98.8% nucleotide identity, encode almost identical WNT receptors with nine amino-acid substitutions. FzE3 is claimed to be expressed specifically in esophageal cancer. We determined the structure of the FZD7 gene and the FZD7 cDNA expressed in esophageal cancer. The FZD7 gene without intron and the FZD7 cDNAs isolated from esophageal cancer cell lines TE4 and TE5 were found to encode WNT receptor identical to FZD7, but not to FzE3. Nucleotide sequence of FzE3 was not identified on the human genome draft sequence. Thus, we could not obtain any data suggesting the existence of FzE3. Expression profile of FZD7 was also investigated. FZD7 was expressed throughout normal gastrointestinal tract, from esophagus to rectum. Among human esophageal and gastric cancer cell lines, expression level of FZD7 was relatively lower in esophageal cancer cell lines, and was highest in the gastric cancer cell line MKN7. FZD7 was up-regulated in one out of six cases of human primary gastric cancer. As over-expression of Frizzled-7 leads to activation of the WNT-beta catenin-TCF pathway, up-regulation of FZD7 in human gastric cancer might play key roles in carcinogenesis through activation of the WNT-beta-catenin-TCF pathway. PMID- 11408931 TI - Reduced expression of REIC/Dkk-3 gene in non-small cell lung cancer. AB - Dickkopfs (Dkks) are secretory glycoproteins and are important for inducing amphibian head formation. Some Dkks have the ability to antagonize the Wnt proto oncoprotein, which can promote cell proliferation and transformation when it is mutated or overexpressed. We recently cloned the REIC/Dkk-3 gene, one of the Dkks, and found that expression of the gene was markedly decreased in many human immortalized and tumor-derived cell lines. To further investigate the function of the REIC/Dkk-3 gene in vivo, the gene expression was quantified in 57 surgically resected non-small cell lung cancer (NSCLC) tissues and their adjacent normal lung tissues by real-time quantitative PCR analysis based on TaqMan methodology. The REIC/Dkk-3 mRNA level was significantly lower in NSCLC tissues than in normal lung tissues. Reduced expression of REIC/Dkk-3 in NSCLC was observed in 36 (63%) of the 57 tumors. Reduced expression of REIC/Dkk-3 was frequently observed in poorly differentiated adenocarcinoma and squamous cell carcinoma. These results indicate that reduced expression of REIC/Dkk-3 may be correlated with rapid cell proliferation in human NSCLC. PMID- 11408932 TI - Molecular cloning and characterization of human WNT8A. AB - WNT - beta-catenin - TCF pathway is involved in carcinogenesis and fetal development. Xenopus wnt-8 is one of the most potent Wnts with the capacity to activate beta-catenin - TCF pathway in the Xenopus axis duplication assay. Here, we have cloned and characterized WNT8A, a novel human homologue of Xenopus wnt-8. The WNT8A gene, consisting of at least 6 exons, was found to encode a 351-amino acid polypeptide with the N-terminal signal peptide, three N-linked glycosylation sites, and conserved amino-acid residues of the WNT family. WNT8A showed 63.2% total-amino-acid identity to WNT8B. C-terminal region of WNT8A, WNT8B, WNT2, WNT2B1 and WNT2B2 were longer than that of other WNTs. Among various normal human tissues and 34 human cancer cells lines, the 3.5-kb WNT8A mRNA was detected only in a human teratocarcinoma cell line NT2. These results strongly suggest that WNT8A might be implicated in development of early embryos as well as germ cell tumors through activation of the WNT - beta-catenin - TCF pathway. PMID- 11408933 TI - Gene microarray analysis reveals a novel hypoxia signal transduction pathway in human hepatocellular carcinoma cells. AB - The molecular details of hypoxia-induced cellular responses have been difficult to identify since there is as yet no known oxygen receptor. We used cDNA microarray technology to extend our studies pertaining to these molecular details in human hepatocellular carcinoma (Hep3B) cells that produce erythropoietin (Epo) in response to hypoxia. Of approximately 1200 genes in the array, those associated with integrin-linked kinase (ILK), fibronectin precursor and glycogen synthase kinase-3beta (GSK-3beta) were markedly stimulated after exposure of Hep3B cells to low oxygen (1%) for 6 h. Epo, HIF-1, and von Hippel-Lindau cDNAs were measured in parallel as markers of low oxygen responses in Hep3B cells. ILK is a serine, threonine protein kinase that interacts with the cytoplasmic domains of integrin beta1 and beta3. This interaction localizes ILK to focal adhesion plaques. ILK is stimulated by cell-fibronectin interaction as well as insulin. It is regulated in a phosphatidylinositol 3-kinase dependent manner and can phosphorylate protein kinase B (PKB/AKT) and GSK-3beta. As a result of these and other activities ILK has been shown to affect anchorage-independent cell survival, cell cycle progression and tumorigenesis in nude mice. ILK has also been implicated in the Wnt pathway and as a critical target in PTEN-dependent tumor therapies. To our knowledge this is the first report implicating the ILK pathway in low oxygen responses. Other genes identified as a result of the microarray analysis not previously known to change as a result of low oxygen treatment were elongation factor-1alpha, glycyl-tRNA synthetase, and laminin receptor protein-1. These findings were all corroborated by RT-PCR assays and in some instances Western blot analysis. PMID- 11408934 TI - Induction of apoptosis by a ribosome-inactivating protein from Agrostemma githago is associated with down-regulation of anti-apoptotic bcl-2 protein expression. AB - Agrostin is a type 1 ribosome-inactivating protein (RIP) isolated from the seeds of Agrostemma githago. 3[H]-thymidine incorporation in human leukemic HL-60 cells was significantly reduced after incubation with agrostin showing that the RIP was inhibitory on the cell proliferation. Results from flow cytometric studies illustrated that apoptosis was induced by agrostin, which was associated with activation of endonucleases and DNA fragmentation of the leukemic cells. Anti apoptotic bcl-2 protein level was also reduced by agrostin showing that the associated apoptotic pathway might involve down-regulation of the intracellular bcl-2 protein level. PMID- 11408935 TI - Clinical significance of plasma nucleosome levels in cancer patients. AB - The nucleosome is the primary repeating unit of DNA organization in chromatin, and cell death may lead to increased levels of circulating nucleosomes in plasma (PNLs) in various circumstances such as inflammation, pulmonary embolism, autoimmune disease, and cancer. Cell death can also be induced by chemotherapeutic agents. We investigated PNLs in 99 patients with primary breast cancer, 26 with recurrent disease, 11 with benign breast disease, and 27 with other histological types of cancer. In 18 patients with recurrent breast cancer who received docetaxel (D, 60 mg/m2) every 3 weeks as second line therapy after an anthracycline-based regimen, PNLs were investigated before and during the administration of D. One hundred and seventy-four healthy controls (111 females, 63 males) without any evidence of disease were also investigated. PNLs were detected using the cell death detection ELISAplus kit. PNLs were significantly higher in patients with primary breast cancer (mean +/- SD: 0.135 +/- 0.213) and in recurrent breast cancer (0.182 +/- 0.196) as compared with healthy female controls (0.010 +/- 0.012) (p < 0.01). In patients with primary breast cancer, no correlation was found between PNLs and clinicopathological characteristics. On the other hand, PNLs were decreased after mastectomy (p < 0.05). Patients with other histological types of cancer (0.244+/-0.383) also showed significantly higher PNLs as compared to healthy controls (p < 0.01), and PNLs were elevated independently of the histological type of cancer. In patients with recurrent breast cancer, PNLs showed a transient increase 24 h after the administration of D, and these increases correlated with the degree of subsequent leukopenia. In a follow-up study, pretreatment baseline PNLs decreased markedly when a response was obtained, whereas there was no decrease in either stable disease or progressive disease. Thus, increased PNLs were found in cancer patients, and PNLs seem to be a sensitive marker of cell death that could be predictive of both leukopenia and response to chemotherapy. PMID- 11408936 TI - Non-Hodgkin's lymphoma presenting with spinal involvement: the Sheffield Lymphoma Group experience (1970-2000). AB - Spinal non-Hodgkin's lymphoma is rare. We retrospectively reviewed the clinical and histopathologic records of 39 consecutive patients referred to the Sheffield Lymphoma Group from 1970 to 2000 and analysed the prognostic differences between localised (stage IE and IIE) and secondary (stage III and IV) spinal non Hodgkin's lymphoma (S-NHL) patients. Forty-five percent of all patients were over 60 years old. More patients were male (58%); presented with stage IE and IIE (63%), mostly of intermediate/high grade histology (74%); over a third had symptoms; nearly a third (11 patients) were paraplegic and 14 had sphincter dysfunction at diagnosis. The overall survival of all patients was 39% at 5 years (median 24.7 months), whilst that of localised S-NHL was 51% (median 89.7 months). Univariate analysis showed better survival for patients with good mobility status at presentation (p < 0.0l) and complete response to initial treatment (p < 0.00l). In primary S-NHL, histology (p < 0.05) significantly influenced overall survival. In conclusion, disease is frequently locally advanced at presentation with aggressive histologic grade: thorough staging should always be performed to exclude widespread disease. Good mobility status predicts for good survival outcome. Optimal treatment is still uncertain. PMID- 11408937 TI - Interference of gemcitabine triphosphate with the measurements of deoxynucleotides using an optimized DNA polymerase elongation assay. AB - The main mechanism of action of the anticancer drug gemcitabine is assumed to be incorporation of its triphosphate (dFdCTP) into DNA, resulting in inhibition of DNA polymerization, inhibition of DNA synthesis and repair. Another mechanism is inhibition of ribonucleotide reductase leading to imbalance in the deoxyribonucleotide (dNTP) pools. One assay to measure dNTP pools is based on oligonucleotide elongation mediated by DNA polymerase. Since the latter may be affected by dFdCTP, we studied the effect of 0.1-600 pmol dFdCTP on this assay; 10 pmol and more dFdCTP significantly increased the average dpm of the blank (absence of other dNTP) and that of the calibration line of dATP (1.4-1.6-fold); 0.1 pmol and more increased that of the standard dGTP curve significantly (1.1 1.8-fold); 10-75 pmol decreased that of dCTP while 75 and 100 pmol significantly increased that of dCTP (1.3-fold); 50 pmol significantly increased that of dTTP (1.3-1.5-fold). For dATP, dGTP and dTTP, a saturation was reached at 100 pmol dFdCTP, but not yet for dCTP. To minimize these effects, we added an excess of 200 pmol dFdCTP to all samples and calibration lines when measuring dNTP levels of gemcitabine treated samples. In this way the effects of gemcitabine on dNTP levels were studied in human A2780 ovarian, HT29 colon, K562 myelogenous leukemia, H322 non-small cell lung cancer cell lines and the murine lung cancer cell line Lewis Lung. In all cell lines, intrinsic dTTP pools (3-77 pmol/106 cells) were the highest, followed by dATP (1.5-31), dCTP (0.7-27) and (nd-14) dGTP. Exposure to 1 and 10 microM gemcitabine for 4-h concentration dependently decreased dATP 3-10-fold and dGTP to undetectable levels, but dCTP at most 3 fold, while dTTP increased. In conclusion, dFdCTP affects dNTP measurements with the DNA polymerase elongation assay, but its effect could be controlled by addition of similar amounts of dFdCTP to each assay. PMID- 11408938 TI - Differential expression of the lung resistance-related protein/major vault protein in the histological compartments of nephroblastomas. AB - Nephroblastomas (Wilms' tumors) are curable with survival rates above 80%. Some tumors, however, fail to respond to therapy and those patients have a poor prognosis. In a search for molecular markers of drug resistance, we investigated the expression of lung resistance protein (LRP) in tissue samples from 32 children with nephroblastoma by means of immunohistochemistry. LRP is a human major vault protein (MVP) and is associated with multidrug resistance of tumors. LRP/MVP expression was found in the blastemal and epithelial compartments but to a significantly lesser extent in the stromal compartment of Wilms' tumors. Expression was generally heterogeneous with respect to staining intensity and percentage of positive cells. We found significant relationships between LRP/MVP expression and chemotherapeutic pre-treatment of tumors and tumor stage. The immunohistochemical results were validated with a real-time RT-PCR technique and a significant association between protein and mRNA expression was observed. PMID- 11408939 TI - Down-regulation of the desmosomal cadherin desmocollin 3 in human breast cancer. AB - In previous studies using cDNA microarray analysis, we have identified an expressed sequence tag which is consistently down-regulated in six human breast tumor cell lines. In the current study, we have determined this tag to be part of the mRNA sequence of human desmocollin 3, a member of the cadherin superfamily of proteins and an integral component of desmosomes. Desmosomes are sites of adhesion between adjacent cells in layers of epithelia, as well as in some non epithelial tissues, and play an important role in the maintenance of tissue structure. Northern analysis, quantitative real-time polymerase chain reaction assay and Western blot analysis showed that desmocollin 3 is present in normal and immortalized human mammary epithelial cells, but consistently exhibits a significant, and often complete, down-regulation in breast cancer cell lines and primary breast tumors, both at the mRNA and protein levels. PMID- 11408940 TI - How surgeon age affects post-treatment surveillance strategies for melanoma patients. AB - The intensity of post-treatment melanoma patient follow-up varies widely among physicians. We investigated whether physician age accounts for the observed variation in surveillance intensity among plastic surgeons. A custom-designed questionnaire was mailed to USA and non-USA surgeons, all of whom were members of the American Society of Plastic and Reconstructive Surgeons. Subjects were asked how they use 14 specific follow-up modalities during years 1-5 and 10 following primary treatment for patients with cutaneous melanoma. Repeated-measures analysis of variance was used to compare practice patterns by TNM stage, year post-surgery, and age. Of the 3,032 questionnaires mailed, 1,142 (38%) were returned. Of those returned, 395 (35%) were evaluable. Non-evaluability was usually due to lack of melanoma patient follow-up in surgeons' practices. Follow up strategies for most of the 14 modalities were highly correlated across TNM stages and years post-surgery, as expected. The pattern of testing varied significantly by surgeon age for 3 modalities (complete blood count, liver function tests, and chest X-ray), but the variation was quite small. We concluded that the post-treatment surveillance practice patterns of ASPRS members caring for patients with cutaneous melanoma vary only marginally with physician age. Continuing medical education could account for this observation. PMID- 11408941 TI - Evidence of a functional role for the cyclin-dependent kinase-inhibitor p21WAF1/CIP1/MDA6 in promoting differentiation and preventing mitochondrial dysfunction and apoptosis induced by sodium butyrate in human myelomonocytic leukemia cells (U937). AB - The impact of dysregulation of the cyclin-dependent kinase inhibitor p21WAF1/CIP1/MDA6 has been examined in U937 human monocytic leukemia cells in relation to cell cycle arrest and differentiation following treatment with the histone deacetylase inhibitor sodium butyrate (SB). Cells stably transfected with a p21WAF1/CIP1/MDA6 antisense construct, in marked contrast to their wild-type counterparts, failed to up-regulate p21WAF1/CIP1/MDA6, undergo G1 arrest, or express the maturation marker CD11b when exposed to 1 or 3 mM SB. However, antisense-expressing cells were significantly more susceptible to SB-mediated mitochondrial injury and apoptosis, manifested by increased cytosolic translocation of cytochrome c, activation of pro-caspase 3, and degradation of PARP. Dysregulation of p21WAF1/CIP1/MDA6 did not modify the extent of SB-induced histone acetylation, but did result in cleavage of p27KIP1, Bcl-2 and pRb, as well as diminished levels of full-length underphosphorylated pRb. Finally, dysregulation of p21WAF1/CIP1/MDA6 did not modify SB-mediated down-regulation of E2F-1 or c-Myc, but was associated with enhanced down-regulation of cyclins D1 and E. Together, these findings indicate that in U937 leukemia cells, p21WAF1/CIP1/MDA6 plays a critical functional role in SB-mediated G1 arrest and maturation, and suggest that cells displaying dysregulation of this CDKI respond to SB by engaging a default apoptotic program. PMID- 11408942 TI - UCN-01 dose-dependent inhibition of normal hyperproliferative cells in mice. AB - UCN-01 is a hydroxylated derivative of staurosporine and a potent protein kinase C (PKC) inhibitor. Interest in the potential usefulness of this compound as an anticancer drug stems mainly from its unique anti-signaling, growth-arresting properties on tumor cells. This include activation of CDC2 kinase (CDK1) which interacts with either cyclin A or cyclin B1 at the G1 or G2/M border, suggeting that this event is one of the major consequences of the drug action on eukaryotic cells. Nonetheless, the antiproliferative activity of UCN-01 on normal rapidly dividing cells (intestinal epithelial and bone marrow cells) is not well documented. Thus, the main objective of this study was to investigate the in vivo antiproliferative activity of UCN-01 on these normal hyperproliferative cells and evaluate whether cellular response to UCN-01 could be modulated in the presence of DNA damage. Mice were injected i.m. with a single dose of UCN-01 (2.5 mg/kg-20 mg/kg) followed 3 and 24 h later by in vivo BrdU labeling for 1 h. At autopsy, bone marrow cells were collected and fixed for dual parameter BrdU/DNA flow cytometry. Different regions of the gut were also fixed for immunoperoxidase BrdU assays. Newly replicated cells were mainly located in the lower compartments of the crypt columns and were scored for BrdU stained nuclei using an image analysis system. A comparison between groups showed that 5 mg/kg UCN-01 induced inhibition in BrdU incorporation at 3 and 24 h, as compared to the other groups injected with various doses of UCN-01. Flow cytometric analysis of bone marrow cells stained with fluorescein tagged anti-BrdU (FITC) along with propidium iodide (PI) also showed inhibition in BrdU incorporation of S phase fraction cells in mice treated with 5 mg/kg UCN-01. These bone marrow cells were arrested primarily in the G1 phase of the cell cycle. The colony-forming unit (CFU) assay of the bone marrow cells was then used to determine the level of drug interaction of UCN-01 and, topotecan, a topoisomerase I inhibitor, at a fixed dose ratio. An antagonistic drug interaction (CI > 1) was observed as determined by the median effect analysis. However, an additive interaction (CI = 1) was obtained with the use of camptothecin or 10,11-methylenedioxycamptothecin and UCN-01. The results of the in vitro drug interaction with UCN-01 may predict protection from topotecan-induced bone marrow toxicity. PMID- 11408943 TI - Mechanism of cardioprotection by resveratrol, a phenolic antioxidant present in red wine (Review). AB - Coronary heart disease (CHD) has been and remains a major contributor to morbidity and mortality in developed countries. The most common form of CHD in the western world is atherosclerosis (AS), especially of the major coronary arteries. Failure to maintain an intact endothelium, as a result of episodic and/or persistent injury and perturbation of the vascular endothelium, promotes formation of fatty streaks which are considered initiation events of AS. Cellular constituents contributing to endothelial injury include endothelial cells, monocytes, platelets, and smooth muscle cells. Individuals diagnosed with AS face complex, enduring clinical complications and enormous medical costs. Simple and easily compliant prevention and treatment measures are therefore strategic considerations in the management of this vascular disease. Based on known risk factors for CHD, priorities in AS prevention should include smoking cessation, blood pressure control, and diet modification. In recent years, the possible benefits of low to moderate consumption of alcoholic beverages, particularly of red wine, in the prevention of heart disease has received increasing attention and debate in the popular media as well as in the scientific community. Such attention has been prompted by research findings supporting a relationship between red wine consumption and the French paradox. This phenomenon refers to people residing in certain parts of France where red wine is customarily consumed during meals having a low CHD mortality, despite living a lifestyle considered to have comparably high CHD risks, as those in the US and many other developed countries. Studies have reported that the cardioprotective effects of red wine are greater than those attributed solely to ethanol and other types of alcoholic beverages. The mechanism(s) underlying the greater CHD protective benefits of red wine have not been elucidated. Recently the polyphenol resveratrol (3,5,4' trihydroxy-trans-stilbene), known to be abundantly present in red wine, compared to white wine, beer, or spirits, has been demonstrated to elicit a broad spectrum of biological responses in in vitro and in animal studies, including effects that are compatible with the cardioprotective roles proposed for red wine. These recently described effects of resveratrol will be reviewed in this article. We will first summarize published data showing an inverse association between consumption of alcoholic beverages/red wine and risk of CHD. A review of biosynthesis of resveratrol and its presence in food groups and wines will follow. Recent studies relating exposure to wine/resveratrol with reduction in myocardial damage during ischemia-reperfusion, modulation of vascular cell functions, inhibition of LDL oxidation, and suppression of platelet aggregation will be presented. The last section of this review will focus on a discussion of mechanism(s) by which resveratrol acts as a potential cardioprotective agent. PMID- 11408944 TI - Long term inhibition of neointima formation in balloon-injured rat arteries by intraluminal instillation of a matrix-targeted retroviral vector bearing a cytocidal mutant cyclin G1 construct. AB - Restenosis from neointimal proliferation is a frequent complication of intracoronary stenting and catheter-based revascularization procedures. Currently, there is no known therapeutic strategy that has been sufficiently effective to warrant its widespread use. In the present study, the anti proliferative properties of a matrix (collagen)-targeted retroviral vector bearing a mutant cyclin G1 (DNT 41-249) construct was evaluated in vitro and in vivo. In controlled one-month efficacy studies, the intraluminal instillation of the mutant cyclin G1 vector significantly inhibited neointima lesion formation in balloon-injured rat arteries without neointimal growth, associated necrosis or intense inflammatory reaction. Taken together, these data extend the potential utility of the matrix-targeted mutant cyclin G1 retroviral vector for management of vascular restenosis. PMID- 11408945 TI - Cyclooxygenase-2 expression in normal ovaries and epithelial ovarian neoplasms. AB - Cyclooxygenase-2 (COX-2) expression was investigated immunohistochemically in 57 epithelial ovarian neoplasms and in histologically normal ovaries. Positive immunostaining for COX-2 was observed in 78.6% (22/28) of the ovarian cancers and in 66.7% (14/21) of the borderline-malignant tumors. The rate of expression was significantly higher among the ovarian cancers than the benign cystadenomas (4/8; 50%) (p<0.05). There was a significant correlation between vascular endothelial growth factor (VEGF) expression and microvessel count (MVC), but no correlation between COX-2 expression and MVC. There was a significant correlation between VEGF expression and COX-2 expression in all of the ovarian neoplasms as a whole (p<0.05). These findings suggest that an increase in COX-2 expression may be associated with malignant transformation and tumorigenesis of epithelial ovarian neoplasms. PMID- 11408946 TI - Uptake of D-mannoheptulose by rat erythrocytes, hepatocytes and parotid cells. AB - D-[3H]mannoheptulose (or D-[1-14C] mannoheptulose) net uptake was measured in rat erythrocytes, parotid cells and hepatocytes. In the erythrocytes and parotid cells, the intracellular distribution space of the heptose (0.1 mM) represented only about 1 and 13%, respectively, of the intracellular 3HOH space. In hepatocytes, however, it amounted to approximately 45% of the intracellular 3HOH space. In all cases, the apparent distribution space of D-[3H]mannoheptulose hexaacetate largely exceeded that of unesterified D-[3H]mannoheptulose. Relative to the intracellular water space, the generation of acidic metabolites (expressed as an apparent distribution space) from radioactive D-mannoheptulose was one order of magnitude lower in parotid cells (< or = 3%) than in hepatocytes (> or = 20%). These findings are compatible with the hypothesis that D-mannoheptulose is transported into cells mainly, if not exclusively, at the intervention of GLUT2. PMID- 11408947 TI - Karyotype analysis of tumorigenic human bronchial epithelial cells transformed by chrysolite asbestos using chemically induced premature chromosome condensation technique. AB - We examined karyotypic changes of tumorigenic human bronchial epithelial cell lines transformed by asbestos fibers. Using Calyculin A mediated premature chromosome condensation (PCC) assay and Giemsa-trypsin banding, we showed that the common changes of all tumorigenic cell lines were the loss of one or two copies of chromosome 5, the monosomy of chromosome 19 and the increased trisomy of chromosome 8. The results indicate that the karyotypic change of chromosome 5, 8 and 19 could play an important role in asbestos-induced tumorigenic conversion of human bronchial epithelial cells from an immortalized to tumorigenic state. PMID- 11408948 TI - Mutational analysis of the Smad6 and Smad7 genes in hepatocellular carcinoma. AB - Resistance to the transforming growth factor-beta (TGF-beta) is a frequently found phenotype in human malignancies. The recent identification of Smad6 and Smad7, both anti-Smads which inhibit TGF-beta signaling, raises a possibility that constitutive activation of the anti-Smads by a somatic mutation may impair the TGF-beta signaling pathway. We tested this hypothesis by screening the entire coding sequences of these anti-Smads for mutations in 52 hepatocellular carcinoma (HCC) samples using polymerase chain reaction - single strand conformation polymorphism analysis. We detected no mutations, but found 3 single nucleotide polymorphisms (SNPs) in the Smad6 gene and 2 SNPs in the Smad7 gene. These results suggest that mutations of the Smad6 and Smad7 genes are not the main cause of the TGF-beta resistance in human HCC. PMID- 11408949 TI - Case populations must match the respective disease model: Genotype diversity causes linkage disequilibrium mapping failure in monogenic disorders. AB - Traditional linkage analysis in large families is the most promising approach for mapping disease genes of monogenic heritable disorders when the number of informative meioses is sufficient. With rare diseases, however, the low availability of informative pedigrees poses a significant limitation. As an adjunct to family linkage methods, association studies based on the investigation of individual haplotypes from a number of unrelated patients (i.e. linkage disequilibrium analysis) have recently been employed in mapping hereditary disease loci. However, such haplotype analysis is hampered by a number of effects that influence statistical evaluation, e.g. i) population history and size, ii) allele and haplotype frequencies in the respective population(s), iii) heterogeneous mutation and natural selection processes, and iv) small sample sizes of patient groups. The purpose of the present study was to determine the utility and limitations of haplotype-based genetic mapping in estimating the location of the NYX gene, which has recently been identified as the causative gene for a rare inherited retinal disorder known as the complete type of X-linked congenital stationary night blindness (CSNB1). For this purpose we recapitulated haplotypes and tested for linkage disequilibrium in 20 unrelated male CSNB1 patients from three European populations and 44 healthy individuals. All subjects were genotyped for 17 polymorphic microsatellite loci covering the Xp11.4 region with an average marker density of approximately 0.29 cM. We found that a precise model to describe mutations at loci that erroneously break up linkage is highly required, and that the case population must match the respective disease model. PMID- 11408950 TI - Inhibitory effect of lactoperoxidase-generated hypothiocyanite upon black pigmented anaerobe growth. AB - This study aimed to evaluate the in vitro effect of lactoperoxidase with or without its substrates (hydrogen peroxide, thiocyanate) on the growth of 4 different black pigmented anaerobe (BPA) strains associated with the development and progress of periodontal diseases: Porphyromonas gingivalis ATCC 33277, Prevotella intermedia NCTC 9336, Prevotella loescheii ATCC 15930, and Prevotella melaninogenica NCTC 9338. A 5-min lactoperoxidase-generated OSCN--HOSCN incubation at pH 6.0, 7.0 or 8.0 resulted in a decrease of the growth rate (tested by turbidimetry in liquid cultures) of the 4 BPA strains, whilst lactoperoxidase alone actually promoted bacterial growth. PMID- 11408951 TI - Glycogen accumulation in cultured tumoral or normal pancreatic islet and acinar cells. AB - Tumoral pancreatic islet cells of either the RINm5F or INS-1 cell lines, when cultured in the presence of 30.0 mM D-glucose, accumulate about 50 times more glycogen than tumoral pancreatic acinar cells of the AR42J line cultured under the same experimental conditions. Expressed per nl of intracellular water, the glycogen content of the RINm5F or INS-1 cells is even higher than that found in rat pancreatic islets also cultured under the same experimental conditions. Moreover, at variance with normal islet cells, tumoral islet cells do not require to be exposed to a high concentration of D-glucose to accumulate large amounts of glycogen. Based on these findings, it is proposed that the labelling of the glycogen pool, e.g. by 11C-labelled D-glucose or 2-deoxy-2-[18F]fluoro-D-glucose, may allow identification and localization of insulinomas in the pancreatic gland by a non-invasive imaging procedure. PMID- 11408952 TI - Expression of p73 and p63 proteins in pancreatic adenocarcinoma: p73 overexpression is inversely correlated with biological aggressiveness. AB - The significance of p53, a prominent tumor suppressor gene, and its product in various neoplasms including pancreatic adenocarcinoma has been established. We investigated the expression of two types of homologue of p53, p73 and p63, in pancreatic adenocarcinoma by means of immunohistochemistry. Overexpression of p73 was observed in 45.6% of the cases and this phenomenon was more frequently seen in cystic adenocarcinomas than in ductal carcinomas. Furthermore, p73 overexpression was inversely linked to lymph node metastasis, tumor size, and Ki 67 labeling index. On the other hand, p63 overexpression was observed in 68.2% of the cases, but was not related to any clinicopathological features. No correlation was established between the expression of these proteins and aberrant p53 expression. These results suggest that decreased expression of p73 protein and over-expression of p63 protein may play a role in the development of pancreatic adenocarcinoma. PMID- 11408953 TI - Cloning and expression of hamster telomerase catalytic subunit cDNA. AB - Topical application of 7,12-dimethylbenz(alpha)-anthracene induces tumors in the hamster cheek pouch. Telomerase activity is increased in the cancer tissues if compared to normal adjacent tissues in both human and hamster oral cancer. In order to achieve a probe and to investigate the putative role of telomerase in oral carcinogenesis using the hamster cheek pouch model, we have cloned the cDNA encoding the hamster telomerase catalytic subunit (hamTERT). The hamster TERT cDNA encoded 1128 amino acids and shared 64% amino acid identity with human TERT and 80% with murine TERT. As noted with human TERT which express several alternatively spliced mRNAs, we have also detected one alternatively spliced hamTERT mRNA in hamster cancer cells. Transient transfection of hamTERT cDNA in a retroviral expression vector reconstituted telomerase activity in the telomerase negative human lung fibroblast IMR90 cells. PMID- 11408954 TI - Genetic polymorphisms and transcriptional pattern analysis of CYP1A1, AhR, GSTM1, GSTP1 and GSTT1 genes in breast cancer. AB - In order to detect the contribution of cytochrome P450 1A1 (CYP1A1), aryl hydrocarbon receptor (AhR), glutathione S-transferases M1 (GSTM1), P1 (GSTP1), and T1 (GSTT1) genes in breast cancer, genetic analysis was performed, as well as transcriptional analysis in sporadic primary tumours and corresponding adjacent normal tissues from the same patient. CYP1A1 3'-untranslated region (3'-UTR) termed as m1 (MspI) polymorphism and the null(-) deletions of both GSTM1 and GSTT1 genes were examined in genomic DNA from blood samples of 207 female breast cancer patients and 171 age and sex matched controls. The frequencies of the m1 genotype of the CYP1A1 gene in cases and controls were 0.13 and 0.15, while the frequencies of homozygotes with GSTM1(-) were 0.52, in each, and for homozygotes with GSTT1(-) were 0.14 and 0.10, respectively. Statistical analysis of these genotypes in combinations did not reveal any significant difference between the breast cancer population and the control group. Expression of mRNA levels of CYP1A1, GSTM1, GSTP1, GSTT1 and AhR genes in 31 breast cancer patients, revealed inter-individual variation in an independent manner to patient age, genotype, or tumour stage. Eighty-seven percent of the tumour specimens tested were deregulated, compared to their normal counterparts, in at least one locus. Up regulation of CYP1A1 was observed only when one of the GSTM1 or GSTP1 was down regulated while the other remained constant. Genotyping analysis did not show any correlation to breast cancer risk. However, RT-PCR analysis provided evidence that CYP1A1, AhR, GSTM1, GSTP1 and GSTT1 genes are frequently deregulated in breast cancer and could be used as molecular biomarkers for better clinical management of such patients, with respect to chemotherapy. PMID- 11408955 TI - Unchanged frequency of loss of heterozygosity and size of the deleted region at 8p21-23 during metastasis of lung cancer. AB - The genetic mechanisms involved in lung cancer development and progression are beginning to be understood. Many studies have documented frequent loss of heterozygosity (LOH) at specific chromosomal regions in cancer cells; this implies that tumor suppressor genes (TSG) are usually present in those regions. Recently, it has been reported that LOH or chromosomal deletions at chromosome 8p21-23 represent early events frequently occurring in lung cancer. In addition, the size of these chromosome 8 deletions, as well as their frequency, was also reported to increase during lung cancer progression. To determine the spectrum and frequency of alterations of chromosome 8p21-23 in human lung cancer and whether these increase with progression of the tumors, we performed LOH analysis of chromosome 8p and 3p in the genomic DNA from cells from primary and metastatic sites of lung cancer, as well as from normal lung. We studied 35 subjects with primary lung cancer including 30 tumors with distant metastasis. Detection of allelic deletion utilized a PCR-based approach of microsatellite polymorphism analysis, which was performed using the microsatellite markers D8S1130, D8S1106, D8S511, D8S1827, D8S549, D8S261, LPL, D8S258, D8S136, NEFL, D3S1295, D3S1313, D3S1234, D3S1300, D3S1351, D3S1339, and D3S1340. The overall allelic deletion rates were 10 of 28 (35.7%) at 8p and 13 of 33 (39.4%) at 3p. The allelic deletions in the primary cancer and its metastatic sites were in each case identical in both frequency and size of the deleted regions. In our analysis, 8p21-23 deletions were not always associated with 3p deletions in primary lung cancer. These results therefore suggest that allelic deletion at chromosome 8p21 23 is an early and frequent event in the carcinogenesis and development of lung cancer, independent of chromosome 3p deletion. However, a continuing increase in the frequency of LOH at 8p21-23 and in the size of the deleted region rarely occurs during the process of metastasis. PMID- 11408956 TI - Comparison of RANTES chemokine induction by Th1 cytokines in human astroglial cell lines. AB - Multiple sclerosis (MS) is a primary inflammatory demyelinating disease of the human central nervous system, characterized by accumulation of mononuclear cells of hematogenous origin. RANTES is a C-C (beta)-chemokine family member with strong chemoattractant activity for lymphocytes and monocytes that are implicated in the pathogenesis of MS lesions. However, the cellular sources of RANTES message and the regulation of its secretion within diseased brain are poorly understood. Therefore, we carried out this study to compare the effect of Th1 cytokines on the induction of RANTES in different human astrocytic cell lines. IFN-gamma alone had little effect on RANTES production in both U-373MG and U 105MG cells, while TNF-alpha or IL-1beta alone had differential effects in the two cell lines. Low levels of RANTES chemokine were detected in culture supernatants from U-373MG cells. By contrast, TNF-alpha or IL-1beta dramatically increased RANTES secretion in U-105MG cells. Interestingly, different combination treatments of cells with the three cytokines synergistically induced RANTES release from both U-373MG and U-105MG cells. Consistent with these results, we found similar expression patterns for RANTES at the comparable steady-state mRNA levels in both cell lines. Furthermore, we showed that U-105MG cells treated with TNF-alpha and IL-1beta alone or in combination markedly induced increases in the rate of transcription of the RANTES chemokine gene. Our results indicate that these cell lines may be good model systems for studying the regulation of RANTES expression by cytokines in human glial cells. PMID- 11408957 TI - Divergent effects of an alpha2-adrenergic antagonist on lipolysis and thermogenesis: interactions with a beta3-adrenergic agonist in rats. AB - This study was undertaken in order to test the hypothesis that selective beta3-AR stimulation and simultaneous blockade of alpha2-AR would result in an increase of lipolysis and thermogenesis in rats. Incubation of isolated white adipocytes with the alpha2-AR antagonist yohimbine produced a concentration-dependent increase in glycerol release (P<0.001) for all assayed concentrations (10-12-10-6 M) and potentiated the lipolytic effect of the beta3-AR agonist Trecadrine. However, in vivo administration of yohimbine produced a marked decrease in body temperature (1.3-1.5 degrees C, P<0.001) and blocked the thermogenic effect of Trecadrine when simultaneously administered. A similar response was observed for whole body oxygen consumption. Furthermore, yohimbine did not modify brown adipose tissue oxygen consumption, but blocked the beta3-AR-mediated increase triggered by Trecadrine. Brown adipose tissue UCP-2 and -3 mRNA expression was not changed by yohimbine. In conclusion, the present work indicates that in vitro alpha2-AR blockade by yohimbine potentiates the beta3-AR-mediated stimulation of lipolysis. On the other hand, in vivo alpha2-AR antagonism blocks the thermogenic effects mediated by beta3-AR stimulation, suggesting a possible interplay between the receptors. PMID- 11408958 TI - Effects of world trade on public health. PMID- 11408959 TI - Properties of olive oil. PMID- 11408960 TI - Prevalence of mental retardation and developmental disabilities: estimates from the 1994/1995 National Health Interview Survey Disability Supplements. AB - In 1994 and 1995, the National Health Interview Survey included a Disability Supplement (NHIS-D) to collect extensive information about disabilities among individuals sampled as part of annual census-based household interview surveys. Here we describe the development and application of operational definitions of mental retardation and developmental disabilities to items in the NHIS-D to estimate prevalence. In our analyses, we estimate the prevalence of mental retardation in the noninstitutionalized population of the United States to be 7.8 people per thousand (.78%); of developmental disabilities, 11.3 people per thousand (1.13%); and the combined prevalence of mental retardation and/or developmental disabilities to be 14.9 per thousand (1.49%). Differences in prevalence estimates for mental retardation and developmental disabilities and among people of various ages are explored. PMID- 11408961 TI - 27th Annual Meeting European Group for Blood and Marrow Transplantation and 17th Meeting of the Nurses Group. Maastricht, The Netherlands, March 25-28, 2001. Abstracts. PMID- 11408962 TI - Tauroursodeoxycholic acid for the cytoprotection of liver grafts during cold storage: a new aspect of its anti-apoptotic properties? PMID- 11408963 TI - Prospects for genetic manipulation of donor organs and tissues. PMID- 11408964 TI - Mechanism and stimulus for fat deposition. PMID- 11408965 TI - Medicaid DSH legislation unveiled on Capitol Hill. PMID- 11408966 TI - Nursing shortage issue garners attention--three bills emerge. PMID- 11408967 TI - Representative Jerry Moran, (R-KS). A health care legislative update from the Chariman of the RHCC. PMID- 11408968 TI - Forceps! Scalpel! Robot! PMID- 11408970 TI - Moms and guilt. PMID- 11408971 TI - Mastering the value chain. An interview with Mark Levin of Millennium Pharmaceuticals. Interview by David Champion. AB - As today's business leaders are all too aware, a new scientific or technological break-through can quickly transform an industry's competitive landscape. The upheaval is often traumatic for the companies involved, forcing them to rethink their strategies and redefine their boundaries. But an industry in flux also creates vast opportunities. To seize them, companies must see how the current upheavals will affect the future distribution of profits--and then reinvent themselves to capitalize on the new sources of value. In this interview with HBR senior editor David Champion, Mark Levin, the founder and CEO of Millennium Pharmaceuticals, describes his vision of the future of the pharmaceutical industry in the wake of the genetics revolution and new technologies that have altered the economics of drug development. No company, he argues, will create serious long-term value by staying in just one or two stages of the value of chain. That's why Millennium, which started out doing basis research into genes and proteins and selling its findings to pharmaceutical giants, has moved downstream - toward the patients who actually use and pay for the drugs. He explains why the research end has become less lucrative than the more mechanical tasks of identifying, testing, and manufacturing molecules. Levin talks about the changes Millennium has undergone since its inception in 1993-from 30 workers to more than 1,000, and from one end of the value chain to the other. He discusses the company's cultural transformations as well as the partnerships and acquisitions that have helped millennium become involved in every stage of the chain-from gene to patient. Levin's vigorous approach to balancing long-term strategy with short-term tactics offers important lessons to any executive facing an industry upheaval. PMID- 11408972 TI - Playing by the rules. How Intel avoids antitrust litigation. AB - Microsoft and Intel are both obvious targets for antitrust litigation; both wield considerable control in their respective segments of the computer industry. But while Mircrosoft has been mired in court for years now--its name and business practices dragged through the mud, and its very future as a single company thrown into doubt--Intel has avoided a prolonged, high-profile antitrust case. Intel's success is not a matter of luck. The company's antitrust compliance program, refined over many years, has been an integral element in the chip maker's business strategy. In this article, the authors suggest that Intel's approach to compliance provides a valuable model for any enterprise that may come under regulators' scrutiny. They describe how Intel created extremely conservative antitrust compliance standards and then instituted a series of unique training events that had active support from then--CEO Andy Grove and others in senior management. First, live training -- not just instructional pamphlets or videos -- was given to all affected employees. Those classes were followed by customized training programs for different parts of the company. Then, to drive antitrust awareness deeper into the company's memory, Intel carried out random audits of employees' files and conducted mock depositions. "It's fascinating to see," Grove says. "A memo is introduced into evidence and you shrug. You fully understand how that memo could be written. Moreover, you could have written it yourself. And then you see that memo turned into a tool and a weapon against you, in front of your eyes." Intel recognizes that no matter how cautious it is, it will always face extraordinary scrutiny as a market leader. But "since antitrust is embedded in everything we do," Grove says, "we can control our destiny." PMID- 11408973 TI - See your brands through your customers' eyes. AB - Subaru markets an L.L. Bean Outback station wagon. Dell stamps Microsoft and Intel logos on its computers. Such inter-weaving of different companies' brands is now commonplace. But one of the central tools of brand management-portfolio mapping--has not kept pace with changes in the marketplace. Most conventional brand maps include only those brands owned by a company, arranged along organizational lines with little regard for how the brands influence customer perceptions. In this article, the authors present a new mapping tool--the brand portfolio molecule--that reveals the way brands appear to customers. The brand portfolio molecule includes all the brands that factor into a consumer's decision to buy, whether or not the company owns them. The first step in creating a brand portfolio molecule is to determine which brands should or should not be included. The second step is to classify each brand by asking five key questions: 1) How important is this brand to customers' purchase decisions about the brand you're mapping? 2) Is its influence positive or negative? 3) What market position does this brand occupy relative to the other brands in the portfolio? 4) How does this brand connect to the other brands in the portfolio? 5) How much control do you have over this brand? The last step is to map the molecule using a 3-D modeling program or by hand with pen and paper. Individual brands take the form of atoms, and they're clustered in ways that reflect how customers see them. The usefulness of the tool lies in its ability to show the many forces that influence a customer's buying decision--and to provide a powerful new way to think about brand strategy. PMID- 11408974 TI - When business is a confidence game. AB - When it comes to making business decisions, being overconfident about your choices can actually be more harmful than just guessing. Here's how managers can calibrate their confidence levels-and avoid being too sure in the wrong situations. PMID- 11408975 TI - A smarter way to buy. AB - Few companies can pinpoint just how much they're spending on procurement. Here's a matrix that gives companies a systematic way to identify and organize the full costs of their relationships with their suppliers. PMID- 11408976 TI - The leader as lobbyist. AB - Guidant's Ginger Graham argues that senior executives are actually better prepared than most lobbyists to inform and educate members of Congress about the issues that will affect their businesses. PMID- 11408977 TI - When your culture needs a makeover. AB - In 1994, the consumer products company Alberto-Culver North America faced flattened sales and the most difficult competitive environment of its history. President Carol Bernick knew that overcoming theses challenges would require a certain kind of corporate culture-but it wasn't the culture the company had. She changed that by focusing on four areas. First, Bernick made culture visible and elevated it to priority status, often by highlighting desired values and behaviors that already existed in pockets of the company. Her annual "state of the company" address stresses that people must be attuned to business realities and the drivers of success. Employees now develop statements of individual economic value describing their contributions to the company's profitability. The company's ten cultural imperatives are required knowledge for all. Second, Bernick and her executive team created the role of growth development leader (GDL). Each mentors about a dozen people. Her frequent meetings with GDLs make them effective agents of change upward and downward. The role is coveted because GDLs have real power to make change: every year they vote on the issues, large and small, that must be addressed by the business as a whole. Third, the company uses an employee survey to identify areas for improvement and to provide 360 degree feedback to GDLs and top management. And fourth, successes are celebrated constantly--through stock awards for the best GDLs, Business Builders Awards for great innovators, and many other, less formal means. Since 1994, the company has cut employee turnover in half, seen sales grow 83%, and watched pretax profits rise 336%--indicators of how cultural change is driving business results. PMID- 11408978 TI - The earnings game. Everyone plays, nobody wins. AB - Quarterly earnings numbers dominate the decisions of executives, analysts, investors, and auditors. Yet for all the attention paid to these numbers, they're not much use in predicting a company's future performance and cash flows. Even economists are unanimous in their view that these numbers say next to nothing about a company's prospects beyond the next quarter. Nonetheless, meetings analysts' expectations that earnings will rise in a smooth, steady, unbroken line has become, at many corporations, a game whose imperatives override even the imperative to deliver the highest possible return to shareholders. The fetishistic attention paid to this almost meaningless indicator might be cause for amusement, except for one thing: the earnings game does real harm. It distorts corporate decision making. It reduces securities analysis and investing to a guessing contest. It compromises the integrity of corporate audits. Ultimately, it undermines the capital markets. As market participants increasingly come to view the quarterly number as a sort of collective fiction, offered and received in a spirit of mutual cynicism, they lose faith in the numbers affected by quarterly earnings--including stock prices themselves. And no market can survive long if its participants see no connection between prices and the intrinsic value of the goods on offer. In this article, HBR senior editor Harris Collingwood takes an in-depth look at these effects, examining the intricacies of the earnings game and why companies believe they have no choice but to play it. Until more corporate executives change their practices, he explains, the earning game will never lack for players. PMID- 11408979 TI - Want to perfect your company's service? Use behavioral science. AB - It may seem like the topic of service management has been exhausted. Legions of scholars and practitioners have applied queuing theory to bank lines, measured response times to the millisecond, and created cults around "delighting the customer." But practitioners haven't carefully considered the underlying psychology of service encounters--the feelings that customers experience during these encounters, feelings often so subtle they probably couldn't be put into words. Fortunately, behavioral science offers new insights into better service management. In this article, the authors translate findings from behavioral science research into five operating principles. First, finish strong: the ending is far more important than the beginning of an encounter because it's what remains in the customer's memory. Second, get the bad experiences out of the way early: in a series of events, people prefer to have undesirable events come first and to have desirable events come last. Third, segment the pleasure, combine the pain: since experiences seem longer when they are broken into segments, it's best to combine all the boring or unpleasant steps of a process into one. Fourth, build commitment through choice: people are happier when they believe they have some control over a process, particularly an uncomfortable one. And fifth, give people rituals and stick to them: most service--encounter designers don't realize just how ritualistic people are. Ultimately, only one thing really matters in a service a encounter--the customer's perception of what occurred. This article will help you engineer your service encounters to enhance your customers' experiences during the process as well as their recollections of the process after it is completed. PMID- 11408980 TI - How to invest in social capital. AB - Business runs better when people within a company have close ties and trust one another. But the relationships that make organizations work effectively are under assault for several reasons. Building such "social capital" is difficult in volatile times. Disruptive technologies spawn new markets daily, and organizations respond with constantly changing structures. The problem is worsened by the virtuality of many of today's workplaces, with employees working off-site or on their own. What's more, few managers know how to invest in such social capital. The authors describe how managers can help their organizations thrive by making effective investments in social capital. For instance, companies that value social capital demonstrate a commitment to retention as a way of limiting workplace volatility. The authors cite SAS's extensive efforts to signal to employees that it sees them as human beings, not just workers. Managers can build trust by showing trust themselves, as well as by rewarding trust and sending clear signals to employees. They can foster cooperation by giving employees a common sense of purpose through good strategic communication and inspirational leadership. Johnson & Johnson's well-known credo, which says the company's first responsibility is to the people who use its products, has helped the company in time of adversity, as in 1982 when cyanide in Tylenol capsules killed seven people. Other methods of fostering cooperation include rewarding the behavior with cash and establishing rules that get people into the habit of cooperating. Social capital, once a given in organizations, is now rare and endangered. By investing in it, companies will be better positioned to seize the opportunities in today's volatile, virtual business environment. PMID- 11408981 TI - Moving upward in a downturn. AB - As the recent bursting of the new economy bubble has shown, business cycles are still wih us. The question, then, is, what executives should do to help their companies weather these downturns. As in so many instances, there are conventional approaches that appear to make sense in the short term. But while these approaches seem reasonable in the heat of the moment, they can eventually damage competitive positions and financial performance. Drawing on extensive research of Fortune 500 companies that have lived through industry downturns and economic recessions over the past two decades, Darrell Rigby, a director of Bain & Company, reveals how companies need to go against the grain of convention and exploit industry downturns to harness their unique opportunities for upward mobility. The author explains that every downturn goes through three phases. He examines each phase and shows how successful players navigate the huge waves of a downturn. Smart executives, he says, don't panic: they look bad news in the eye and institutionalize an approach to detecting storms. Rather than hedge their bets through diversification, they focus on their core businesses and spend to gain market share. They manage costs relentlessly during good times and bad. They keep a long-term view and strive to maintain the loyalty of employees, suppliers, and customers. And coming out of the downturn, they maintain momentum in their businesses to stay ahead of the competition they've already surpassed. Every industry will face periodic downturns of varying severity, says Rigby. But executives with the vision and ingenuity to take unconventional approaches can buoy their companies to new heights. PMID- 11408982 TI - Dalteparin as periprocedure anticoagulation for patients on warfarin and at high risk of thrombosis. AB - BACKGROUND: Patients taking warfarin and at high risk for thromboembolic complications have traditionally been hospitalized for two to three days to receive standard treatment with intravenous heparin both prior to and following procedures while their international normalized ratio (INR) is subtherapeutic. OBJECTIVE: To assess the feasibility of protocol implementation for outpatient anticoagulation with low-molecular-weight heparin to eliminate or reduce the length of hospital admission needed solely for anticoagulation. METHODS: Patients included were receiving warfarin for a prosthetic heart valve, mitral valve disease with atrial fibrillation, or recent episode of venous thromboembolism. Warfarin was discontinued four days prior to the procedure. Subcutaneous dalteparin 200 units/kg was given on the two mornings prior to the procedure and restarted 12-24 hours after the procedure until the INR was in the therapeutic range. Warfarin was reinitiated on the evening of surgery. RESULTS: Twenty-four patients underwent 26 procedures. There were two minor bleeding complications, and one patient experienced a transient ischemic attack. Patients received a median of five days of dalteparin. The INR returned to the therapeutic range on the median postoperative day 4. All patients avoided two days of hospitalization prior to the procedure (i.e., no patients needed to be admitted preoperatively for anticoagulation). A median of four days would have been required for the sole purpose of postoperative anticoagulation. CONCLUSIONS: Outpatient perioperative anticoagulation with dalteparin for high-risk patients requiring long-term oral anticoagulation appears feasible and warrants further study. PMID- 11408983 TI - Topical phenytoin treatment of stage II decubitus ulcers in the elderly. AB - OBJECTIVE: To compare the healing of stage II decubitus ulcers with topically applied phenytoin sodium with two other standard topical treatment procedures in a long-term care setting; and to assess the extent of systemic absorption after topical application in the phenytoin group. METHODS: Forty-seven nursing home patients with stage II decubitus ulcers were chosen for this study. The patients were matched for age, gender, and size and severity of wounds, and randomly assigned to each treatment group. Clinical assessment of decubitus ulcers was performed at the beginning of treatment and at each dressing change. Ulcers were examined for the presence of healthy granulation tissue, reduction in surface dimensions, and time to healing. Two phenytoin sodium plasma concentrations were to be obtained on all patients in the phenytoin group. RESULTS: Topical phenytoin therapy resulted in a shorter time to complete healing and formation of granulation tissue when compared with DuoDerm dressings or triple antibiotic ointment applications (p < or = 0.05). The mean +/- SD time to healing in the phenytoin group was 35.3 +/- 14.3 days compared with 51.8 +/- 19.6 and 53.8 +/- 8.5 days for the DuoDerm and triple antibiotic ointment groups, respectively. Healthy granulation tissue in the phenytoin group appeared within two to seven days in all subjects. Patients in the standard treatment groups required six to 21 days to produce new granulation tissue. Serum phenytoin sodium concentrations were nondetectable. No patient withdrew from the study secondary to adverse treatment effects. CONCLUSIONS: Both the phenytoin and standard treatment groups showed progress over the study period. However, the phenytoin group demonstrated more rapid results in all aspects of ulcer healing. PMID- 11408984 TI - Medicaid's beta 2-agonist recipients and their treatment by national standards. AB - OBJECTIVE: To determine whether Connecticut's Medicaid high-dose beta 2-agonists (HDB) users (> 1 canister/mo) are receiving medications according to National Institutes of Health (NIH) asthma guidelines, and to compare healthcare utilization between HDB users and patients receiving no more than one canister of a short-acting beta 2-agonist per month. METHODS: All Connecticut Medicaid pharmacy claims from April to December 1998 were examined. Subjects were included if they had an asthma diagnosis and were excluded if they had chronic obstructive pulmonary disease. The percentage of HDB users not receiving long-term-control medication; receiving low doses of a long-term-control medication; and receiving oral, nebulized, or long-acting beta 2-agonists was determined. Healthcare utilization between HDB users and patients receiving no more than one canister of a short-acting beta 2-agonist per month was also compared. RESULTS: Of 1599 profiles included, 14% (n = 223) were HDB users. Twenty-five percent of these patients did not receive a long-term-control medication, and 35% received low doses of a long-term-control medication. Claims for oral, nebulized, and long acting beta 2-agonists were submitted for 4%, 29%, and 22% of HDB users, respectively. More claims for short courses of oral corticosteroids were submitted for HDB users than for patients receiving no more than one canister of a short-acting beta 2-agonist per month (n = 1376) (1.99 +/- 3.60 vs. 0.39 +/- 1.07 mean +/- SD; p < 0.001). HDB users visited their physicians' offices an average of 4.72 +/- 24.08 times per month compared with an average of 2.40 +/- 14.4 office visits per month for patients receiving no more than one canister of a short-acting beta 2-agonist (p = 0.04). CONCLUSIONS: A high proportion of Connecticut's Medicaid HDB users are not receiving medications according to NIH guidelines. HDB users were significantly more likely to receive short courses of oral corticosteroids and required significantly more office visits than patients receiving no more than one canister of a short-acting beta 2-agonist per month. Intervention programs designed to improve adherence to the guidelines will be developed. PMID- 11408985 TI - Levofloxacin failure in a patient with pneumococcal pneumonia. AB - OBJECTIVE: To report a case of levofloxacin failure in a patient with a penicillin-sensitive Streptococcus pneumoniae pneumonia. CASE SUMMARY: A previously healthy, immunocompetent, 53-year-old white man presented with penicillin-sensitive S. pneumoniae pneumonia. The patient was empirically placed on levofloxacin monotherapy, which was continued due to a local penicillin shortage. When the patient failed to improve, further susceptibility testing was ordered. The organism was found to have a penicillin minimum inhibitory concentration (MIC) of 0.023 microgram/mL and a levofloxacin MIC of 6 micrograms/mL. Effective antimicrobial therapy was delayed, as clinicians did not anticipate fluoroquinolone resistance. DISCUSSION: Newer fluoroquinolones such as levofloxacin have good activity against most S. pneumoniae isolates and are used for the treatment of pneumonia. Although resistance to these agents is rare, it has been reported. Current guidelines from the National Committee for Clinical Laboratory Standards do not recommend initial fluoroquinolone susceptibility testing. CONCLUSIONS: As fluoroquinolone resistance may not be identified by susceptibility patterns to other antibiotics, early fluoroquinolone susceptibility testing and increased awareness of resistance may aid clinicians in their treatment of pneumococcal disease. PMID- 11408986 TI - Effect of levofloxacin on theophylline clearance during theophylline and clarithromycin combination therapy. AB - OBJECTIVE: To report a case of decreased theophylline clearance by the addition of levofloxacin in a patient receiving theophylline and clarithromycin. CASE SUMMARY: A 59-year-old Japanese man who was receiving theophylline for emphysema experienced stimulation, insomnia, and tachycardia due to theophylline toxicity after clarithromycin and levofloxacin were added to the regimen. The combination of these agents resulted in a decrease in theophylline clearance to approximately 60% of the initial value obtained while the patient was receiving theophylline alone. The adverse effects disappeared after the dosage was reduced and the theophylline serum concentration decreased; however, there was no change in theophylline clearance. After discontinuation of levofloxacin, the theophylline serum concentration decreased, and theophylline clearance returned to the initial level even though clarithromycin was continued. DISCUSSION: Levofloxacin is believed not to influence the clearance of theophylline, although some new fluoroquinolones have been reported to do so. This case indicates that levofloxacin and clarithromycin inhibited theophylline metabolic pathways catalyzed by both CYP1A2 and CYP3A4 and resulted in the decrease in theophylline clearance. The clearance of theophylline, therefore, is not influenced by clarithromycin alone. CONCLUSIONS: Careful monitoring is required when levofloxacin is prescribed for patients who are taking clarithromycin with theophylline. PMID- 11408987 TI - Trimethoprim/sulfamethoxazole-induced toxic epidermal necrolysis. AB - OBJECTIVE: To report a case of toxic epidermal necrolysis (TEN) associated with trimethoprim/sulfamethoxazole (TMP/SMX). CASE SUMMARY: A 34-year-old Asian woman developed a severe, desquamating mucocutaneous reaction (TEN) after six days of taking TMP/SMX to treat a presumed urinary tract infection (UTI). DISCUSSION: TMP/SMX is often recommended as first-line therapy for UTIs, sinusitis, bronchitis, and as prophylaxis and treatment for Pneumocystis carinii pneumonia. TEN is a rare, but severe condition associated with sulfonamide use. This article describes a typical case and offers an opportunity for review of this potentially serious reaction. CONCLUSIONS: Sulfonamides are often implicated in the majority of drug-induced cases of TEN. This case report illustrates the typical presentation of sulfonamide-induced TEN with a prodrome, characteristic rash, mucous membrane lesions, and systemic involvement. Practitioners should be aware of this rare adverse effect and closely observe patients for cutaneous manifestations or complaints. Any suspected drug should be discontinued if clinical evaluation leads to the suspicion of Stevens-Johnson syndrome or TEN. PMID- 11408988 TI - Neuroleptic malignant syndrome during a change from haloperidol to risperidone. AB - OBJECTIVE: To report a case of neuroleptic malignant syndrome (NMS) in a patient whose therapy was being switched from haloperidol to risperidone. CASE REPORT: A 57-year-old African-American man, treated for schizophrenia with haloperidol for several years, developed NMS within 48 hours of the addition of low doses of risperidone and mirtazapine to his regimen. Symptoms, which included fever, generalized rigidity, and altered mental status, resolved after discontinuation of psychotropics, supportive management, and several weeks of treatment with bromocriptine and dantrolene. He was subsequently treated with olanzapine without adverse effects. DISCUSSION: Several cases of NMS have been reported with risperidone, but none under these circumstances. NMS most likely occurred in this patient as a result of the additive dopamine 2 receptor blocking of haloperidol and risperidone. Sympathetic hyperactivity secondary to mirtazapine may also have been a contributing factor. If NMS may be induced by the simultaneous use of older, high-potency antipsychotics and newer, atypical antipsychotics such as risperidone, switching patients from older to newer antipsychotics may at times be difficult, since completely stopping one antipsychotic before starting the second may place patients at risk for psychotic relapse. CONCLUSIONS: Clinicians should closely monitor patients receiving both haloperidol and risperidone or combinations of similar medications. PMID- 11408989 TI - Relief of fibromyalgia symptoms following discontinuation of dietary excitotoxins. AB - BACKGROUND: Fibromyalgia is a common rheumatologic disorder that is often difficult to treat effectively. CASE SUMMARY: Four patients diagnosed with fibromyalgia syndrome for two to 17 years are described. All had undergone multiple treatment modalities with limited success. All had complete, or nearly complete, resolution of their symptoms within months after eliminating monosodium glutamate (MSG) or MSG plus aspartame from their diet. All patients were women with multiple comorbidities prior to elimination of MSG. All have had recurrence of symptoms whenever MSG is ingested. DISCUSSION: Excitotoxins are molecules, such as MSG and aspartate, that act as excitatory neurotransmitters, and can lead to neurotoxicity when used in excess. We propose that these four patients may represent a subset of fibromyalgia syndrome that is induced or exacerbated by excitotoxins or, alternatively, may comprise an excitotoxin syndrome that is similar to fibromyalgia. We suggest that identification of similar patients and research with larger numbers of patients must be performed before definitive conclusions can be made. CONCLUSIONS: The elimination of MSG and other excitotoxins from the diets of patients with fibromyalgia offers a benign treatment option that has the potential for dramatic results in a subset of patients. PMID- 11408990 TI - Mifepristone. AB - OBJECTIVE: To review the efficacy and safety of mifepristone (with misoprostol) for the termination of early pregnancy. DATA SOURCES: A MEDLINE search (1966 October 2000) was conducted, and additional references listed in articles were included; unpublished data obtained from the manufacturer were used to identify data from the scientific literature. Studies evaluating mifepristone were considered for inclusion. STUDY SELECTION: Human clinical studies in the English language were reviewed and evaluated. Clinical trials selected for detailed review were limited to those including the regimens of mifepristone and misoprostol, recently approved by the Food and Drug Administration for early pregnancy termination. DATA SYNTHESIS: Mifepristone is an antiprogestin available for pregnancy termination in combination with a prostaglandin such as misoprostol. Mifepristone offers efficacy similar to, if not better than, other drugs used for pregnancy termination, but appears less efficacious overall than surgical termination of pregnancy. Mifepristone in combination with misoprostol commonly causes adverse effects such as abdominal pain and, less commonly, can cause serious adverse effects such as incomplete abortion; endometritis; and bleeding warranting transfusion, hospitalization, or surgery. Mifepristone is metabolized by the cytochrome P450 system. Thus, the potential for drug interactions with this agent exists, although this has not been well studied. Data are included from clinical trials evaluating the safety, tolerability, efficacy, and pharmacoeconomics of mifepristone combined with misoprostol for early pregnancy termination. Data comparing the use of these agents with surgical abortion and other drugs used for pregnancy termination are included where available. CONCLUSIONS: Mifepristone in combination with misoprostol for the termination of early pregnancy (amenorrhea of < or = 49 d) is effective in 92-95% of women. Incomplete abortion requiring surgical abortion after the fact occurs in 3-5% of women, and pregnancy continues 1-2% of the time. Mifepristone with misoprostol treatment is not without significant risks, including hemorrhage, infection, and potential for long-term emotional consequences. PMID- 11408991 TI - Intravenous itraconazole. AB - OBJECTIVE: To review the pharmacology, mycology, chemistry, pharmacokinetics, efficacy, safety, tolerability, dosage, administration, and economic issues of intravenous itraconazole. DATA SOURCES: A MEDLINE search from 1978 to June 2000 of the English-language literature and an extensive review of meeting abstracts was conducted. Due to the paucity of published information concerning the pharmacokinetics, efficacy, and safety of the intravenous formulation of intravenous itraconazole, additional information was obtained from the manufacturer. DATA EXTRACTION: Data from in vitro and preclinical studies, as well as Phase II and III clinical trials, were included. DATA SYNTHESIS: The triazole antifungal agent itraconazole is available in a cyclodextrin-based intravenous formulation. Intravenous itraconazole is indicated for the treatment of pulmonary and extrapulmonary blastomycosis; histoplasmosis, including chronic cavitary pulmonary disease and disseminated, nonmeningeal histoplasmosis; and pulmonary and extrapulmonary aspergillosis in patients who are intolerant of or who are refractory to amphotericin B. This formulation provides quicker and more consistent therapeutic concentrations than the oral formulations. Clinical data comparing the efficacy of intravenous itraconazole with that of amphotericin B are lacking. CONCLUSIONS: Intravenous itraconazole offers a less toxic alternative for patients with pulmonary and extrapulmonary blastomycosis, histoplasmosis, and aspergillosis who cannot receive oral medications or who are intolerant of or refractory to amphotericin B. PMID- 11408992 TI - Proton-pump inhibitors for acute peptic ulcer bleeding. AB - OBJECTIVE: To review the use of proton-pump inhibitors for acute peptic ulcer bleeding. DATA SOURCES: Articles were obtained through computerized searches of MEDLINE (1966-September 2000). Additionally, several textbooks containing information on the diagnosis and management of acute peptic ulcer bleeding were reviewed. The bibliographies of retrieved publications and textbooks were reviewed for additional references. STUDY SELECTION: All randomized studies and pharmacoeconomic evaluations that used proton-pump inhibitor therapy for acute peptic ulcer bleeding were included. Randomized controlled trials and meta analyses involving other therapies for treating peptic ulcer bleeding were also reviewed for possible inclusion. DATA EXTRACTION: The primary outcomes extracted from the literature were persistent or recurrent bleeding, transfusion requirements, need for endoscopic intervention or surgery, length of stay, and mortality. DATA SYNTHESIS: Data from double-blind, placebo-controlled trials involving more than 1000 patients demonstrate that short-term, high-dose omeprazole therapy is effective for reducing bleeding and transfusion requirements in patients with acute peptic ulcer bleeding. The patients most likely to benefit from this therapy are hospitalized patients at high risk for rebleeding and patients in whom endoscopic evaluation must be delayed or is unavailable. CONCLUSIONS: Omeprazole (and likely other proton-pump inhibitors) is useful in reducing bleeding and transfusion requirements in patients with acute peptic ulcer bleeding, although better delineation of appropriate candidates is needed. PMID- 11408993 TI - Use of cidofovir in progressive multifocal leukoencephalopathy. AB - BACKGROUND: Progressive multifocal leukoencephalopathy (PML) is a subacute demyelinating illness caused by the JC virus, a polyomavirus that occurs in 4-5% of HIV-positive patients. Mortality is high, and no useful therapy has been identified. Highly active antiretroviral therapy (HAART) has been reported to be effective in halting progression of the disease in some, but not all, patients. Cidofovir has been shown to be active against polyomaviruses. OBJECTIVE: To review data on the use of cidofovir to treat PML. DATA SOURCES: English-language case reports and clinical studies were located through a literature search (MEDLINE and AIDSLINE, 1995-July 2000). STUDY SELECTION AND DATA EXTRACTION: Relevant case reports and studies describing the use of cidofovir for PML were reviewed. DATA SYNTHESIS: Most case reports describing the use of cidofovir have shown that the drug is effective in the treatment of PML. Some patients were also receiving HAART concurrently; therefore, it is not clear which treatment modality had a greater impact on PML. However, cidofovir may be effective in patients whose disease has progressed despite HAART or who are-unable to tolerate these regimens. A pilot study of cidofovir for treating PML has completed enrollment, but preliminary results showed no benefit. CONCLUSIONS: Cidofovir may be the most reasonable treatment option for PML in HIV-infected individuals who fail to improve with HAART or who are unable to tolerate these regimens. Patients who receive cidofovir should be monitored for renal and ocular toxicity. PMID- 11408994 TI - Role of therapeutic drug monitoring for protease inhibitors. AB - OBJECTIVE: To review the role of therapeutic drug monitoring (TDM) for protease inhibitors in the treatment of HIV infection. DATA SOURCES: Primary articles were identified using MEDLINE (1966-October 2000), EMBASE (1987-October 2000), AIDSLINE (1980-October 2000), Current Contents, PubMed, and Medscape. Further articles were identified from bibliographic review of primary articles and review papers. Abstracts presented at the World AIDS Conference, Interscience Conference on Antimicrobial Agents and Chemotherapy, and Conference on Retroviruses and Opportunistic Infections were also identified from 1997 to 2000. STUDY SELECTION AND DATA EXTRACTION: All English-language, prospective clinical trials, as well as selected retrospective studies and case series, pertaining to therapeutic drug monitoring of protease inhibitors were included. DATA SYNTHESIS: A number of clinical studies have found a good relationship between concentration and pharmacologic response and/or toxicity as well as wide interpatient variability in the pharmacokinetics of protease inhibitors. There also is some preliminary evidence of the usefulness of plasma drug concentrations to guide dosage adjustments of protease inhibitors in patients with liver dysfunction. Furthermore, there is preliminary evidence of a relationship between drug concentrations and resistance. CONCLUSIONS: A number of clinical studies support the usefulness of TDM of protease inhibitors. However, before TDM can be of the most value, further evaluation requires simplified and standardized assays to be performed routinely by clinical laboratories; determination of the appropriate target concentration and therapeutic range, as well as the best predictor of pharmacologic response; and refined interpretation of plasma drug concentrations. Randomized, controlled clinical trials of patient outcomes are needed to assess the clinical utility of TDM for protease inhibitors. PMID- 11408995 TI - L-arginine in the management of cardiovascular diseases. AB - OBJECTIVE: To examine the role of L-arginine in the management of cardiovascular diseases. DATA SOURCES: A MEDLINE search (1966-April 2000) of review articles, using the search terms arginine, nitric oxide, and cardiovascular diseases, was conducted. After reviewing these articles, primary studies using the search terms arginine, hypercholesterolemia, hypertension, diabetes, smoking, ischemic heart disease, and heart failure were reviewed. STUDY SELECTION: English-language human studies were selected and evaluated based on quality of review. DATA SYNTHESIS: Small-scale studies have demonstrated that intravenous L-arginine augments endothelial function by enhancing vasodilation and reducing monocyte adhesion. Oral supplementation demonstrated similar effects as well as improvement of exercise ability in patients with cardiovascular diseases. CONCLUSIONS: L arginine improves the management of multiple cardiovascular diseases. However, most published human studies are small. Before therapy can be routinely recommended, larger, well-designed studies are required to confirm its effect. PMID- 11408996 TI - Adjunctive rifampin therapy for central nervous system staphylococcal infections. AB - OBJECTIVE: To review the published clinical data assessing the role of adjunctive rifampin therapy for the treatment of staphylococcal central nervous system (CNS) infections. DATA SOURCES: A MEDLINE search (January 1966-March 2000) of English language literature pertaining to CNS staphylococcal infections and rifampin was performed; tertiary sources were also used. DATA EXTRACTION: Human data and case reports were included, as no clinical trials have been published. DATA SYNTHESIS: Retrospective reviews of rifampin used in combination with other antibiotics for serious bacterial infections show conflicting results. Few case reports have described clinical successes with adjunctive rifampin therapy for CNS infections. CONCLUSIONS: The routine use of adjunctive rifampin for CNS infections cannot be justified. PMID- 11408997 TI - Writing manuscripts describing clinical trials: a guide for pharmacotherapeutic researchers. AB - Once researchers have completed a clinical trial and analyzed the data, they have a duty to make the results known to their peers by writing a manuscript suitable for publication in a biomedical journal. Before beginning any writing, those involved in the clinical drug trial first need to decide among themselves who is to be the first author and secondary authors, the readership they want to influence, and the most appropriate target journal to reach this readership. For content and formatting of the manuscript, writers need to carefully follow the instructions for authors as stipulated by the target journal. In general, a manuscript based on a clinical trial is presented sequentially in four sections: Introduction, Methods, Results, and Discussion (IMRAD). The introduction concisely presents background information about the topic that was researched in the clinical trial, provides the study hypothesis, and describes the study rationale, purpose, and objectives. The Methods section delineates the study design, subject/patient selection process, procedures, end points, and types of analysis conducted in the clinical trial to investigate the study hypothesis. The Results section presents the findings of the clinical trial, and the Discussion section interprets the meaning of these findings, assesses how the findings answer the study hypothesis, and compares the study data with findings reported in earlier studies. Writers should select a title for the clinical trial manuscript that is fewer than 10 words and which embodies the essence of the study findings clearly and specifically without being sensationalistic. Writers should create the abstract last; the abstract should provide an overview of the manuscript and stimulate readers' interest. Overall, following the above principles in writing a manuscript based on a clinical drug trial will make the actual writing easier and more enjoyable, as well as increase the possibility of the manuscript being accepted for publication. PMID- 11408998 TI - Publication of clinical drug trials. AB - Clinical drug trials are the best means for establishing the efficacy and safety of drug therapy. Peer-reviewed publications are the most efficient and reliable means for wide dissemination of the results. The investigators and industry have an obligation to publish the data obtained from these trials so that patients may receive the most effective treatment in the safest possible way. PMID- 11408999 TI - Possible celecoxib-induced gastroduodenal ulceration. PMID- 11409000 TI - Benzylpenicillin-induced prolonged cholestasis. PMID- 11409001 TI - Neuroleptic malignant syndrome with antidepressant/antipsychotic drug combination. PMID- 11409002 TI - Comment: fluorouracil-induced aphasia: neurotoxicity versus cerebral ischemia. PMID- 11409003 TI - The relationships between personal PM exposures for elderly populations and indoor and outdoor concentrations for three retirement center scenarios. AB - Personal exposures, indoor and outdoor concentrations, and questionnaire data were collected in three retirement center settings, supporting broader particulate matter (PM)--health studies of elderly populations. The studies varied geographically and temporally, with populations studied in Baltimore, MD in the summer of 1998, and Fresno, CA in the winter and spring of 1999. The sequential nature of the studies and the relatively rapid review of the mass concentration data after each segment provided the opportunity to modify the experimental designs, including the information collected from activity diary and baseline questionnaires and influencing factors (e.g., heating, ventilation, and air-conditioning (HVAC) system operation, door and window openings, air exchange rate) measurements. This paper highlights both PM2.5 and PM10 personal exposure data and interrelationships across the three retirement center settings, and identifies the most probable influencing factors. The current limited availability of questionnaire results, and chemical speciation data beyond mass concentration for these studies, provided only limited capability to estimate personal exposures from models and apportion the personal exposure collections to their sources. The mean personal PM2.5 exposures for the elderly in three retirement centers were found to be consistently higher than the paired apartment concentrations by 50% to 68%, even though different facility types and geographic locations were represented. Mean personal-to-outdoor ratios were found to 0.70, 0.82, and 1.10, and appeared to be influenced by the time doors and windows were open and aggressive particle removal by the HVAC systems. Essentially identical computed mean PM2.5 personal clouds of 3 micrograms/m3 were determined for two of the studies. The proposed significant contributing factors to these personal clouds were resuspended particles from carpeting, collection of body dander and clothing fibers, personal proximity to open doors and windows, and elevated PM levels in nonapartment indoor microenvironments. PMID- 11409004 TI - Particulate matter and heart rate variability among elderly retirees: the Baltimore 1998 PM study. AB - This study investigates the relationship between ambient fine particle pollution and impaired cardiac autonomic control in the elderly. Heart rate variability (HRV) among 56 elderly (mean age 82) nonsmoking residents of a retirement center in Baltimore County, Maryland, was monitored for 4 weeks, from July 27 through August 22, 1998. The weather was seasonally mild (63-84 degrees F mean daily temperature) with low to moderate levels of fine particles (PM2.5 < 50 micrograms/m3). Two groups of approximately 30 subjects were examined on alternate days. A spline mixed-effects model revealed a negative relationship between outdoor 24-h average fine particulate matter (PM2.5) and high-frequency (HF) HRV that was consistent with our earlier Baltimore study for all but 2 days. These 2 days were the only days with significant precipitation in combination with elevated PM2.5. They were also unusual in that back-trajectory of their air masses was distinctly different from those on the other study days, emanating from the direction of rural Pennsylvania. Mixed-effects analysis for all 24 study days showed a small negative association of outdoor PM2.5 with HF HRV (-0.03 change in log[HF HRV] for a 10 micrograms/m3 increment in PM2.5) after adjustment for age, sex, cardiovascular status, trend, maximum temperature, average dew point temperature, random subject intercepts, and autocorrelated residuals. After excluding study days 4 and 5, this association was strengthened (-0.07 change in log[HF HRV] for 10 micrograms/m3 PM2.5, 95% CI -0.13 to -0.02) and was similar to that obtained in an earlier study (-0.12 change in log[HF HRV] for a 10 micrograms/m3 increment in outdoor PM2.5, 95% CI -0.24 to -0.00) [Liao D., Cai J., Rosamond W.D., Barnes R.W., Hutchinson R.G., Whitsel E.A., Rautaharju P., and Heiss G. Cardiac autonomic function and incident coronary heart disease: a population-based case-cohort study. The ARIC Study. Atherosclerosis Risk in Communities Study. Am J Epidemiol 1997: 145 (8): 696-706]. Acute (1 to 4 h) previous PM2.5 exposure did not have a stronger impact than the 24-h measure. A distributed lag model incorporating the six preceding 4-h means also did not indicate any effect greater than that observed in the 24-h measure. This study is consistent with earlier findings that exposures to PM2.5 are associated with decreased HRV in the elderly. PMID- 11409005 TI - Experimental methodologies and preliminary transfer factor data for estimation of dermal exposures to particles. AB - Developmental efforts and experimental data that focused on quantifying the transfer of particles on a mass basis from indoor surfaces to human skin are described. Methods that utilized a common fluorescein-tagged Arizona Test Dust (ATD) as a possible surrogate for housedust and a uniform surface dust deposition chamber to permit estimation of particle mass transfer for selected dust size fractions were developed. Particle transfers to both wet and dry skin were quantified for contact events with stainless steel, vinyl, and carpeted surfaces that had been pre-loaded with the tagged test dust. To better understand the representativeness of the test dust, a large housedust sample was collected and analyzed for particle size distribution by mass and several metals (Pb, Mn, Cd, Cr, and Ni). The real housedust sample was found to have multimodal size distributions (mg/g) for particle-phase metals. The fluorescein tagging provided surface coatings of 0.11-0.36 ng fluorescein per gram of dust. The predominant surface location of the fluorescein tag would best represent simulated mass transfers for contaminant species coating the surfaces of the particles. The computer-controlled surface deposition chamber provided acceptably uniform surface coatings with known particle loadings on the contact test panels. Significant findings for the dermal transfer factor data were: (a) only about 1/3 of the projected hand surface typically came in contact with the smooth test surfaces during a press; (b) the fraction of particles transferred to the skin decreased as the surface roughness increased, with carpeting transfer coefficients averaging only 1/10 those of stainless steel; (c) hand dampness significantly increased the particle mass transfer; (d) consecutive presses decreased the particle transfer by a factor of 3 as the skin surface became loaded, requiring approximately 100 presses to reach an equilibrium transfer rate; and (e) an increase in metals concentration with decreasing particle size, with levels at 25 microns typically two or more times higher than those at 100 microns--consistent with the earlier finding of Lewis et al. for the same sample for pesticides and polycyclic aromatic hydrocarbons (PAHs). PMID- 11409006 TI - An assessment of the data quality for NHEXAS--Part I: Exposure to metals and volatile organic chemicals in Region 5. AB - A National Human Exposure Assessment Survey (NHEXAS) was performed in U.S. Environmental Protection Agency (U.S. EPA) Region V, providing population-based exposure distribution data for metals and volatile organic chemicals (VOCs) in personal, indoor, and outdoor air, drinking water, beverages, food, dust, soil, blood, and urine. One of the principal objectives of NHEXAS was the testing of protocols for acquiring multimedia exposure measurements and developing databases for use in exposure models and assessments. Analysis of the data quality is one element in assessing the performance of the collection and analysis protocols used in NHEXAS. In addition, investigators must have data quality information available to guide their analyses of the study data. At the beginning of the program quality assurance (QA) goals were established for precision, accuracy, and method quantification limits. The assessment of data quality was complicated. First, quality control (QC) data were not available for all analytes and media sampled, because some of the QC data, e.g., precision of duplicate sample analysis, could be derived only if the analyte was present in the media sampled in at least four pairs of sample duplicates. Furthermore, several laboratories were responsible for the analysis of the collected samples. Each laboratory provided QC data according to their protocols and standard operating procedures (SOPs). Detection limits were established for each analyte in each sample type. The calculation of the method detection limits (MDLs) was different for each analytical method. The analytical methods for metals had adequate sensitivity for arsenic, lead, and cadmium in most media but not for chromium. The QA goals for arsenic and lead were met for all media except arsenic in dust and lead in air. The analytical methods for VOCs in air, water, and blood were sufficiently sensitive and met the QA goals, with very few exceptions. Accuracy was assessed as recovery from field controls. The results were excellent (> or = 98%) for metals in drinking water and acceptable (> or = 75%) for all VOCs except o-xylene in air. The recovery of VOCs from drinking water was lower, with all analytes except toluene (98%) in the 60-85% recovery range. The recovery of VOCs from drinking water also decreased when comparing holding times of < 8 and > 8 days. Assessment of the precision of sample collection and analysis was based on the percent relative standard deviation (% RSD) between the results for duplicate samples. In general, the number of duplicate samples (i.e., sample pairs) with measurable data were too few to assess the precision for cadmium and chromium in the various media. For arsenic and lead, the precision was excellent for indoor, and outdoor air (< 10% RSD) and, although not meeting QA goals, it was acceptable for arsenic in urine and lead in blood, but showed much higher variability in dust. There were no data available for metals in water and food to assess the precision of collection and analysis. PMID- 11409007 TI - Toxicokinetics of human exposure to methyl tertiary-butyl ether (MTBE) following short-term controlled exposures. AB - Methyl tertiary-butyl ether (MTBE) is an oxygenated compound added to gasoline to improve air quality as part of the US Federal Clean Air Act. Due to the increasing and widespread use of MTBE and suspected health effects, a controlled, short-term MTBE inhalation exposure kinetics study was conducted using breath and blood analyses to evaluate the metabolic kinetics of MTBE and its metabolite, tertiary-butyl alcohol (TBA), in the human body. In order to simulate common exposure situations such as gasoline pumping, subjects were exposed to vapors from MTBE in gasoline rather than pure MTBE. Six subjects (three females, three males) were exposed to 1.7 ppm of MTBE generated by vaporizing 15 LV% MTBE gasoline mixture for 15 min. The mean percentage of MTBE absorbed was 65.8 +/- 5.6% following exposures to MTBE. The mean accumulated percentages expired through inhalation for 1 and 8 h after exposure for all subjects were 40.1% and 69.4%, respectively. The three elimination half-lives of the triphasic exponential breath decay curves for the first compartment was 1-4 min, for the second compartment 9-53 min, and for the third compartment 2-8 h. The half-lives data set for the breath second and blood first compartments suggested that the second breath compartment rather than the first breath compartment is associated with a blood compartment. Possible locations for the very short breath half-life observed are in the lungs or mucous membranes. The third compartment calculated for the blood data represent the vessel poor tissues or adipose tissues. A strong correlation between blood MTBE and breath MTBE was found with mean blood-to breath ratio of 23.5. The peak blood TBA levels occurred after the MTBE peak concentration and reached the highest levels around 2-4 h after exposures. Following the exposures, immediate increases in MTBE urinary excretion rates were observed with lags in the TBA excretion rate. The TBA concentrations reached their highest levels around 6-8 h, and then gradually returned to background levels around 20 h after exposure. Approximately 0.7-1.5% of the inhaled MTBE dose was excreted as unchange urinary MTBE, and 1-3% was excreted as unconjugated urinary TBA within 24 h after exposure. PMID- 11409008 TI - Effect of vehicle use and maintenance patterns of a self-described group of sensitive individuals and nonsensitive individuals to methyl tertiary-butyl ether in gasoline. AB - The objective of this study was to compare the driving habits and vehicle maintenance patterns of individuals who report symptoms when exposed to methyl tertiary-butyl ether (MTBE) and those who are asymptomatic when exposed to the oxygenate. Participants were healthy volunteers (CON) and self-reported MTBE sensitive individuals (SRS) who participated in a controlled exposure study of MTBE in gasoline. A questionnaire was developed to gather information about each participant's automobile usage, engine maintenance habits and fueling and driving patterns. Results showed that the individuals who had self-reported heightened sensitivity to the oxygenate drove their vehicles more often and fueled their vehicles more frequently than asymptomatic individuals. In addition, the self reported symptomatic individuals in this study were shown to be more likely to drive vehicles with some form of body damage and carbureted engines. PMID- 11409009 TI - Correcting for the effects of location and atmospheric conditions on air pollution exposures in a case-crossover study. AB - A limitation of most air pollution health effects studies is that they rely on monitoring data averaged over one or more ambient monitors to represent daily air pollution exposures for individuals. Such data analyses therefore implicitly require the assumption of a homogeneous spatial distribution for particulate matter (PM). This assumption may be suspected in the Pacific Northwest because of its hilly topography and local variations in wood burning. To examine the bias from substituting regional PM (i.e., the average of three ambient monitor measurements) for individual PM exposure, we conducted an exposure substudy to identify the influence of location factors, specifically urban versus suburban classification and topographic features ("upstream" versus "downstream"), on local ambient measurements. Using nephelometer measurements collected over 1 year in four locations, we developed regression models to predict local PM as a function of regional PM, atmospheric stagnation, temperature, and location. We found a significant interaction between atmospheric stagnation and topography, with the most upstream site having reduced PM levels on high stagnation days after controlling for regional PM. We also found a significant interaction with temperature at one downstream site thought to be heavily exposed to wood smoke in the winter. These results are consistent with the physics of surface radiation inversions. The interactions reordered the index versus referent exposures in a case-crossover analysis of out-of-hospital primary cardiac arrest for subjects living in specific locations, but did not meaningfully change the associations with PM from the analysis using regional PM as the exposure. The lack of change in these results may be due to limitations in the data used to correct the exposure estimates or to the absence of a PM effect among persons without prior heart disease who experienced a primary cardiac arrest. PMID- 11409010 TI - Estimation of particulate matter from visibility in Bangkok, Thailand. AB - Lack of daily data on airborne particles has been a common problem in an air pollution research. To deal with this problem, a regression model was developed to estimate daily PM10 concentration using visibility in Bangkok from 1992 to 1997, based on 1092 visibility/PM10 pair-observations on low humidity days (humidity < or = 76.5%). Visibility was significantly and inversely associated with PM10 (r = 0.71), after adjusting for minimum temperature and winter indicator variable. The R2 of the model was 0.51. PMID- 11409011 TI - Human resources. Waste not, want not. PMID- 11409012 TI - Referrals. Risky business. AB - There are wide variations in referral rates between GPs. The reasons for this are not well understood. Reduction in referrals will require protocols and the involvement of patients. PMID- 11409013 TI - Fever pitch. PMID- 11409014 TI - Peripheral vascular disease in Asia: a rapidly changing pattern. PMID- 11409015 TI - Art macabre: is anatomy necessary? PMID- 11409016 TI - Ruptured abdominal aortic aneurysms: clinical presentation in Auckland 1993-1997. PMID- 11409017 TI - Long-term prognosis of femoropopliteal bypass: an analysis of 349 consecutive revascularizations. AB - BACKGROUND: Femoropopliteal bypass is the commonest procedure for lower limb revascularization. The aim of the present study was to determine the long-term outcomes of femoropopliteal bypass and evaluate the prognostic significance of various clinical factors on the long-term results. METHODS: From 1976 to 1998, 349 consecutive primary femoropopliteal bypass operations were performed on 314 patients at the University of Hong Kong Medical Centre. Indications for operation included claudication (n = 85) and limb-threatening ischaemia (n = 264). Univariate and multivariate analyses of 14 clinical variables were undertaken to identify significant prognostic factors affecting the graft patency, limb salvage and patient survival rates. RESULTS: The overall primary patency rates of femoropopliteal bypass were 88%, 79% and 76% at 1, 3 and 5 years, respectively. Type of graft material and age of patient were independent prognostic factors of graft patency. The cumulative limb salvage rates were 90%, 86% and 86% at 1, 3 and 5 years, respectively. No clinical factor was found to be predictive of ultimate limb loss. The overall survival rates were 89%, 85% and 78% at 1, 3 and 5 years, respectively. Coronary artery disease was the main cause of late death. Gender and indication for operation were the significant predictive factors of long-term survival. CONCLUSIONS: Femoropopliteal bypass using reversed long saphenous vein provided the most durable long-term patency. Autologous saphenous vein should be the choice of vascular conduit if available. Male gender and limb threatening ischaemia were associated with a poor survival. PMID- 11409018 TI - Ruptured abdominal aortic aneurysms: clinical presentation in Auckland 1993-1997. AB - BACKGROUND: Rupture of an abdominal aortic aneurysm (RAAA) carries a reported mortality rate in the range of 32-95%. Survival requires prompt diagnosis and surgical management. The presenting features, however, are varied, often insidious and potentially misleading with Osler noting nearly 100 years ago that a correct premortem diagnosis was achieved in only 33% of cases. The present study aims to review our present accuracy in diagnosing this condition and outline demographic and presenting features of patients with RAAA. METHODS: A review was undertaken of hospital and Coroner's files of all patients residing in the Auckland Coronial region who had RAAA between 1 January 1993 and 31 December 1997. RESULTS: Three hundred and twenty-nine cases of RAAA were identified, and they occurred most commonly in the 8th decade. The male:female ratio was 3:1 and at least 73% of patients were Caucasian. The overall mortality was 71%. Nearly half underwent surgery and the hospital averaged mortality rate was 46%. No patient survived without surgery. Classic presenting features of RAAA were absent in many cases. Abdominal pain, back pain and a palpable mass occurred in only 49%, 36% and 18% of patients, respectively. Other common presenting symptoms included vomiting, general malaise and pelvic or hip pain. Forty-three patients (16%) were initially misdiagnosed. CONCLUSIONS: Although our ability to correctly diagnose a RAAA has improved since Osler's time, the initial misdiagnosis rate of 16% leaves no room for complacency. Ruptured abdominal aortic aneurysms must be included in the differential diagnosis of any patient over the age of 55 years who presents with shock, even if the pain is non-specific or atypical. PMID- 11409019 TI - Fine-needle aspiration cytology of parotid tumours: is it useful? AB - BACKGROUND: The efficacy of fine-needle aspiration cytology (FNAC) in the diagnosis of parotid tumours remains a controversial subject. Studies within small departments utilizing experienced pathologists have shown FNAC to have high sensitivity and specificity for parotid tumours. The present study was performed to assess the accuracy and utility of FNAC of parotid tumours within a teaching hospital environment. METHODS: One hundred and sixty-nine patients underwent both FNAC and subsequent surgery to the parotid between 1995 and 1999. The results of the FNAC were compared to the histopathological diagnosis obtained from the surgical specimen. RESULTS: Fine-needle aspiration cytology had an overall accuracy of 56%. Approximately 10% of the FNAC results were non-diagnostic. The sensitivity and specificity for the following diagnoses were, respectively: benign 86% and 61%; malignant 57% and 100%; pleomorphic adenoma 78% and 95%; squamous cell carcinoma 52% and 99%; mucoepidermoid carcinoma 14% and 99% and adenocarcinoma 20% and 100%. Six non-neoplastic conditions were misdiagnosed and all six patients underwent surgery. CONCLUSIONS: Fine-needle aspiration cytology was found to be highly specific for malignancy but its sensitivity for malignancy was poor. The results show that although FNAC is relatively inexpensive and minimally invasive, it cannot be relied upon to provide an accurate tissue diagnosis, may fail to identify malignancy and does not prevent patients undergoing surgery for non-neoplastic conditions. PMID- 11409020 TI - Nephron-sparing surgery for small incidental renal cell carcinoma. AB - BACKGROUND: Nephron-sparing surgery is currently an accepted treatment for renal cell carcinomas in patients with bilateral tumours, solitary kidneys and when overall renal function is impaired or at risk from medical disease. Its role in patients with a normal contralateral kidney remains controversial. METHODS: The authors' experience in 23 patients undergoing partial nephrectomy for small peripheral lesions between 1995 and 2000 is reported here. RESULTS: Twenty-three patients (13 men and 10 women) with a mean age of 56 years underwent partial nephrectomy. All but three of these patients had a normal contralateral kidney. Mean operating time was 141 min with a mean reduction of haemoglobin of 28 g/dL. Three patients required transfusion. Serum creatinine did not change significantly between preoperative and postoperative values. Two JJ stents were placed prophylactically during surgery to minimize urinary leak. There were no intraoperative or early postoperative deaths and at mean follow up of 16 months there was no evidence of recurrent tumour in 23 patients. Seventeen per cent of lesions removed were benign. CONCLUSIONS: Partial nephrectomy for small peripheral lesions is a safe procedure with low morbidity. No definite recurrences are evident at an early stage of follow up, although longer review (probably more than 10 years) will be required to assess cancer-specific survival following this procedure. PMID- 11409021 TI - Synthetic bone graft substitutes. AB - Replacement of extensive local bone loss is a significant clinical challenge. There are a variety of techniques available to the surgeon to manage this problem, each with their own advantages and disadvantages. It is well known that there is morbidity associated with harvesting of autogenous bone graft and limitations in the quantity of bone available. Alternatively allografts have been reported to have a significant incidence of postoperative infection and fracture as well as the potential risk of disease transmission. During the past 30 years a variety of synthetic bone graft substitutes has been developed with the aim to minimize these complications. The benefits of synthetic grafts include availability, sterility and reduced morbidity. The present article examines the relevance of synthetic bone graft substitutes, their mechanical properties and clinical application. PMID- 11409022 TI - Comparison of three methods in surgical treatment of pilonidal disease. AB - BACKGROUND: The present study was designed to compare three methods that are still used for the surgical treatment of pilonidal disease: marsupialization, primary midline closure and skin flaps. METHODS: One hundred and one out of a total of 203 pilonidal disease patients underwent excision and marsupialization, while 82 patients had excision and primary closure and the remaining 20 were treated with excision and skin flaps. The minimum and maximum follow-up periods for the aforementioned surgical methods were 4 and 5 years, respectively. All patients were reviewed for in-hospital stay, return to work, wound infection and recurrence rates. Student's t-test and Fisher's exact test were used for statistical analysis. RESULTS: Average hospital stays for marsupialization, primary closure and skin flaps were 2.84 +/- 0.13, 2.62 +/- 0.12 and 5.95 +/- 0.52 days, respectively. Hospital stay for the skin flaps method was longer than that for the other two methods. The average time to return to work after marsupialization was 5.42 +/- 0.08 weeks; but the time needed to return to work after undergoing the primary closure or the skin flaps methods was much shorter: 2.15 +/- 0.05 and 2.90 +/- 0.20 weeks, respectively (P < 0.001). There was no difference in wound infection rate (P = 1.000) or recurrence rates. CONCLUSION: The fact that there were no differences in terms of wound infection or recurrence rates between the three groups, and the relatively shorter period for returning to work, emphasize the usefulness of the excision and repair techniques in the surgical treatment of pilonidal disease. PMID- 11409023 TI - Minimal residual marrow disease: detection and significance of isolated tumour cells in bone marrow. AB - BACKGROUND: Detection of malignant cells in bone marrow and peripheral blood of patients with solid tumours at the time of surgery is increasingly emerging as a prognostic factor for disease progression and a way of monitoring adjuvant therapies. Furthermore, isolation and characterization of these cells provide insight into the early metastatic process, with potential therapeutic implications. This article reviews the current knowledge about the clinical significance of minimal residual marrow disease (MRMD) and its methods of detection, outlining some of their specific technical problems. METHODS: A comprehensive review of articles cited in the largest medical databases was conducted. RESULTS: The sensitivity of the methods of detecting MRMD has improved substantially in the past decade, resulting in higher detection rates. In many solid tumours MRMD has been found to correlate with disease-free and overall survival; however, the importance of this as an independent prognostic variable remains contentious. CONCLUSIONS: There is a need for a standardized approach to the detection of these cells before they become integrated into the current staging systems. The challenge remains to establish which of these tumour cells have the capacity to progress and develop metastatic disease, what are the early genetic and molecular mechanisms underlying this process and to apply novel, better targeted therapies against them. PMID- 11409025 TI - Current status of sentinel node biopsy in breast cancer. PMID- 11409024 TI - Art macabre: resurrectionists and anatomists. AB - The teaching of anatomy in England and Scotland from the 16th to the 19th centuries was carried out by the Companies of Barber Surgeons and also there were a number of private schools. The only sources of material for dissection and study were the gallows or the grave and the supply from the former was limited by law. Therefore the latter became the source of a saleable commodity, and so the profession of grave robbing became established. The taking of bodies was abhorrent to the populace, fights and riots would sometimes occur and public outrage was directed towards anatomists. The passing of the Anatomy Act of 1832 helped bring an end to the grisly business of snatching bodies, but the supply of material for study still remained a problem. In the 1920s there was a change in public attitude toward dissection which resulted in an increase in the donation of bodies. PMID- 11409026 TI - Wild duck strikes again. PMID- 11409027 TI - Intravesical explosion during transurethral resection of the prostate. PMID- 11409028 TI - Pseudotumour cerebri due to a torcular epidermoid cyst. PMID- 11409029 TI - Advanced renal cell carcinoma in the absence of a mass. PMID- 11409030 TI - Homozygous serum amyloid P component-deficiency does not enhance regression of AA amyloid deposits. AB - Serum amyloid P component (SAP) is a common protein constituent of all types of amyloid deposits. Using SAP-deficient mice generated through gene targeting, we and others have shown that SAP significantly promotes amyloid deposition. It has been speculated that SAP protects amyloid fibrils from degradation by coating their exterior surface. To assess potential ways of treating individuals with amyloidosis, we examined the persistence of splenic AA amyloid fibrils in SAP deficient and wild-type mice. No enhancement in the rate of regression of splenic AA amyloid was observed in the SAP-deficient mice relative to wild-type mice. These results present, for the first time, evidence that lack of SAP in AA amyloid deposits does not enhance regression of the deposits in vivo and suggest that dissociation of bound SAP from AA amyloid deposits would not significantly accelerate regression of the deposits in vivo. PMID- 11409031 TI - Nodular cutaneous amyloidosis and carpal tunnel syndrome due to the amyloidogenic transthyretin His 114 variant. AB - This is the second report of transthyretin (TTR) amyloidosis in a patient who had ATTR Tyr114His diagnosed by mass spectrometry and gene analysis. This case had some clinical features that differed from those of the first reported cases. The patient, 73-year-old man, complained of generalized cutaneous tubercula that had started at age 68. These tubercula gradually increased in size and became generalized. He felt a slight numbness in his extremities. Clinical and electrophysiological examinations revealed that he had bilateral carpal tunnel syndrome (CTS), whereas there was no clear evidence of sensory and/or motor polyneuropathy. Autonomic symptoms were not present. Biopsy studies revealed that both his tuberculum and his sural nerve contained TTR-related amyloid. In his sural nerve, amyloid deposits were observed mainly in the perineurium, not in the endoneurium, and there was no significant depletion of myelinated fibers. The features of this patient were clinically characterized by generalized cutaneous amyloid deposits and late-onset CTS with a lack of overt polyneuropathy and autonomic dysfunction. The unique clinical features in this case seemed to be consistent with the distribution of amyloid deposits. PMID- 11409032 TI - AL 366--a glycosylated protein of kappa 1b origin in a patient with systemic amyloidosis of predominantly non-parenchymatous distribution. PMID- 11409033 TI - Genotypes at SAA1 locus correlate with the clinical severity of AA-amyloidosis. PMID- 11409034 TI - Two Spanish sibs with familial amyloidotic polyneuropathy homozygous for the V30M TTR gene. AB - Two Spanish sibs with familial amyloidotic polyneuropathy (FAP) homozygous for the V30M-TTR gene, were diagnosed by DNA and protein analyses. Their clinical picture was very similar to the Majorcan FAP heterozygous patients except for the sensorimotor syndrome which was more aggressive. Noteworthy were clinical differences between the sibs concerning autonomic involvement, cranial neuropathy and kidney disturbances. These differences can be due to genetic and/or environmental factors. PMID- 11409035 TI - Secretases as therapeutic targets for the treatment of Alzheimer's disease. AB - The extracellular deposition of short amyloid peptides in the brain of patients is thought to be a central event in the pathogenesis of Alzheimer's Disease. The generation of the amyloid peptide occurs via a regulated cascade of cleavage events in its precursor protein, A beta PP. At least three enzymes are responsible for A beta PP proteolysis and have been tentatively named alpha-, beta- and gamma-secretases. The recent identification of several of these secretases is a major leap in the understanding how these secretases regulate amyloid peptide formation. Members of the ADAM family of metalloproteases are involved in the non-amyloidogenic alpha-secretase pathway. The amyloidogenic counterpart pathway is initiated by the recently cloned novel aspartate protease named BACE. The available data are conclusive and crown BACE as the long-sought beta-secretase. This enzyme is a prime candidate drug target for the development of therapy aiming to lower the amyloid burden in the disease. Finally, the gamma secretases are intimately linked to the function of the presenilins. These multi transmembrane domain proteins remain intriguing study objects. The hypothesis that the presenilins constitute a complete novel type of protease family, and are cleaving A beta PP within the transmembrane region, remains an issue of debate. Several questions remain unanswered and direct proof that they exert catalytic activity is still lacking. The subcellular localization of presenilins in neurons, their integration in functional multiprotein complexes and the recent identification of additional modulators of gamma-secretase, like nicastrin, indicate already that several players are involved. Nevertheless, the rapidly increasing knowledge in this area is already paving the road towards selective inhibitors of this secretase as well. It is hoped that such drugs, possibly in concert with the experimental vaccination therapies that are currently tested, will lead to a cure of this inexorable disease. PMID- 11409036 TI - First international conference on cerebral amyloid angiopathy, October 5-6, 2000, Boston, MA. PMID- 11409037 TI - Biophysical analysis of normal transthyretin: implications for fibril formation in senile systemic amyloidosis. AB - Transthyretin (TTR) is a plasma protein that transports thyroid hormone and retinol binding protein-vitamin A complex. Eighty-four variants of TTR have been identified and seventy-four are associated with familial amyloidotic polyneuropathy. Normal TTR is the major protein found in the fibrillar deposits in the heart at time of autopsy of individuals with senile systemic amyloidosis. The mechanism by which normally soluble TTR deposits as organ-damaging, insoluble, pathological fibrils late in life is unknown. Understanding the mechanism of fibrillogenesis of normal TTR is critical to the design of clinical treatments aimed at retardation, prevention, or reversal of fibril deposition. We have employed a biophysical approach to explore the hypothesis that an instability in a particular secondary or tertiary structure plays a role in the ability of normal TTR to form fibrils at physiological pH. Using far UV circular dichroic (CD) spectroscopy as a function of temperature we have identified simultaneous, cooperative, reversible structural changes in the beta-sheet and alpha-helical regions. The flexible short, surface-located loops undergo an irreversible conformational change at a lower temperature. Spectra before and after heating are different, particularly in the wavelength region associated with these loops, strongly suggesting that the major portion of TTR returns to its initial conformation while the loops do not. Near UV CD reveals partially reversible and irreversible changes in tertiary structure. Using calorimetry to directly measure the enthalpy associated with these changes, two peaks are observed, with further analysis suggesting conformational intermediates. Precipitates from heated samples reveal pre-fibrillar morphology by negative stain electron microscopy. These biophysical studies suggest that heat-induced conformational rearrangements enable normal TTR to assemble into pre-fibrils at physiological pH. PMID- 11409038 TI - Abnormal immunoglobulin synthesis in monoclonal immunoglobulin light chain and light and heavy chain deposition disease. AB - The Congo red-binding fibrils of AL amyloidosis are the most common form of monoclonal immunoglobulin tissue deposition (MIDD). Nonetheless, the less structured deposits found in light chain deposition disease (LCDD) and the similar, but distinct, deposits of light and heavy chain deposition disease (LHCDD) and heavy chain deposition disease (HCDD) can produce significant clinical pathology. Analyses of immunoglobulin synthesis by bone marrow cells obtained from 7 patients with LCDD and LHCDD demonstrated the production of excess light chains in all and the presence of incomplete light chains or heavy chain fragments in 5, regardless of the presence of an intact monoclonal protein or related subunit in the serum or urine. Our data indicate that, as is the case with the fibrillar deposits of AL amyloid, the non-fibrillar forms of monoclonal Ig deposition (LCDD and LHCDD) can be associated with the presence of immunoglobulin fragments in bone marrow cells. In some instances these appeared to be synthetic in origin, although rapid intracellular proteolysis or a combination of both could not be excluded. In either case the fragments may be more susceptible to tissue deposition, with subsequent organ compromise, than intact Ig chains. PMID- 11409039 TI - Pore formation by beta-2-microglobulin: a mechanism for the pathogenesis of dialysis associated amyloidosis. AB - Beta-2 microglobulin (beta 2M, molecular weight 10,000) is a 99 residue immune system protein which is part of the MHC Class I complex whose role is to present antigens to T cells. beta 2M serum levels rise dramatically in renal failure, and a syndrome called "dialysis associated amyloidosis" occurs with time in a majority of hemodialysis patients who exhibit beta 2M amyloid deposits in joints, bone and other organs. beta 2M can also induce Ca++ efflux from calvariae, collagenase production, and bone resorption. We report here that beta 2M formed relatively nonselective, long-lived, voltage independent ion channels in planar phospholipid bilayer membranes at physiologically relevant concentrations. The channels were inhibited by Congo red and blocked by zinc suggesting that they exist in an aggregated beta sheet state as is common with other amyloid fibril forming peptides. Multiple single channel conductances were seen suggesting that various oligomers of beta 2M may be capable of forming channel structures. We suggest that beta 2M channel formation may account for some of the pathophysiologic effects seen in dialysis associated amyloidosis. These findings lend further weight to the "channel hypothesis" of amyloid pathogenesis. PMID- 11409040 TI - Systematic reviews. PMID- 11409041 TI - An easy chair with a difference: a home-made appliance. PMID- 11409042 TI - Status quo in the executive suite. Hospital CEO turnover rate stable; most change seen in D.C., southwest states. PMID- 11409043 TI - Three N.E. states to link drug buying. PMID- 11409044 TI - UPMC finds success with health plan. After years of losses, Pittsburgh venture uses docs to help turn profit. PMID- 11409046 TI - Getting in on the inside. 'Hospital-within-a-hospital' idea catching on in N.C. for long-term acute care. PMID- 11409047 TI - Overall, not so bad. Survey finds hospital systems posted a modest operating profit in 2000. PMID- 11409048 TI - Split decision. Nurses lose crucial ruling on union representation, but win right to sue. PMID- 11409049 TI - Heart-healthy. Cardiac-care provider MedCath ready to again tap the IPO vein. PMID- 11409050 TI - Mammalian play: training for the unexpected. AB - In this review, we present a new conceptual framework for the study of play behavior, a hitherto puzzling array of seemingly purposeless and unrelated behavioral elements that are recognizable as play throughout the mammalian lineage. Our major new functional hypothesis is that play enables animals to develop flexible kinematic and emotional responses to unexpected events in which they experience a sudden loss of control. Specifically, we propose that play functions to increase the versatility of movements used to recover from sudden shocks such as loss of balance and falling over, and to enhance the ability of animals to cope emotionally with unexpected stressful situations. To obtain this "training for the unexpected," we suggest that animals actively seek and create unexpected situations in play through self-handicapping; that is, deliberately relaxing control over their movements or actively putting themselves into disadvantageous positions and situations. Thus, play is comprised of sequences in which the players switch rapidly between well-controlled movements similar to those used in "serious" behavior and self-handicapping movements that result in temporary loss of control. We propose that this playful switching between in control and out-of-control elements is cognitively demanding, setting phylogenetic and ontogenetic constraints on play, and is underlain by neuroendocrinological responses that produce a complex emotional state known as "having fun." Furthermore, we propose that play is often prompted by relatively novel or unpredictable stimuli, and is thus related to, although distinct from, exploration. We present 24 predictions that arise from our new theoretical framework, examining the extent to which they are supported by the existing empirical evidence and contrasting them with the predictions of four major alternative hypotheses about play. We argue that our "training for the unexpected" hypothesis can account for some previously puzzling kinematic, structural, motivational, emotional, cognitive, social, ontogenetic, and phylogenetic aspects of play. It may also account for a diversity of individual methods for coping with unexpected misfortunes. PMID- 11409051 TI - The origin of the attine ant-fungus mutualism. AB - Cultivation of fungus for food originated about 45-65 million years ago in the ancestor of fungus-growing ants (Formicidae, tribe Attini), representing an evolutionary transition from the life of a hunter-gatherer of arthropod prey, nectar, and other plant juices, to the life of a farmer subsisting on cultivated fungi. Seven hypotheses have been suggested for the origin of attine fungiculture, each differing with respect to the substrate used by the ancestral attine ants for fungal cultivation. Phylogenetic information on the cultivated fungi, in conjunction with information on the nesting biology of extant attine ants and their presumed closest relatives, reveal that the attine ancestors probably did not encounter their cultivars-to-be in seed stores (von Ihering 1894), in rotting wood (Forel 1902), as mycorrhizae (Garling 1979), on arthropod corpses (von Ihering 1894) or ant faeces in nest middens (Wheeler 1907). Rather, the attine ant-fungus mutualism probably arose from adventitious interactions with fungi that grew on walls of nests built in leaf litter (Emery 1899), or from a system of fungal myrmecochory in which specialized fungi relied on ants for dispersal (Bailey 1920) and in which the ants fortuitously vectored these fungi from parent to offspring nests prior to a true fungicultural stage. Reliance on fungi as a dominant food source has evolved only twice in ants: first in the attine ants, and second in some ant species in the solenopsidine genus Megalomyrmex that either coexist as trophic parasites in gardens of attine hosts or aggressively usurp gardens from them. All other known ant-fungus associations are either adventitious or have nonnutritional functions (e.g., strengthening of carton-walls in ant nests). There exist no unambiguous reports of facultative mycophagy in ants, but such trophic ant-fungus interactions would most likely occur underground or in leaf litter and thus escape easy observation. Indirect evidence of fungivory can be deduced from contents of the ant alimentary canal and particularly from the contents of the infrabuccal pocket, a pharyngeal device that filters out solids before liquids pass into the intestine. Infrabuccal pocket contents reveal that ants routinely ingest fungal spores and hyphal material. Infrabuccal contents are eventually expelled as a pellet on nest middens or away from the nest by foragers, suggesting that the pellet provides fungi with a means for the dispersal of spores and hyphae. Associations between such "buccophilous" fungi and ants may have originated multiple times and may have become elaborated and externalized in the case of the attine ant-fungus mutualism. Thus, contrary to the traditional model in which attine fungi are viewed as passive symbionts that happened to come under ant control, this alternative model of a myrmecochorous origin of the attine mutualism attributes an important role to evolutionary modifications of the fungi that preceded the ant transition from hunter-gatherer to fungus farmer. PMID- 11409052 TI - Perspectives. A lot to learn, a lot to pay for in nursing education. PMID- 11409053 TI - AIDS at 20. PMID- 11409054 TI - Can he find a cure? PMID- 11409055 TI - The angry prophet is dying. PMID- 11409056 TI - A 20-year toll. PMID- 11409057 TI - 'Our house is on fire!'. PMID- 11409058 TI - Surviving sleepovers. PMID- 11409059 TI - Correlates of actual and perceived knowledge of prostate cancer among African Americans. AB - While digital rectal examination, prostate-specific antigen, and transrectal ultrasound have been identified as effective means of early detection of prostate cancer, African American men tend to underuse these services as compared to white men. Using a nonrandom sample of 108 African American men, the authors conducted an exploratory investigation of the effects of education, income, age, and health insurance coverage on actual and perceived knowledge of prostate cancer. The extent to which the use of prostate cancer screening services may be attributed to actual and perceived knowledge of prostate cancer was also explored. Respondents demonstrated a poor knowledge of prostate cancer and less than 40% reported having had prostate cancer screening as part of their annual physical examination. The results of the study also revealed that (a) there was a moderately strong correlation between actual and perceived knowledge of prostate cancer, (b) use of prostate cancer screening service was positively associated with actual and perceived knowledge of the disease, (c) actual knowledge of prostate cancer was negatively correlated with education, age, and income, and (d) actual and perceived knowledge of prostate cancer were both correlated with having health insurance coverage. PMID- 11409060 TI - Patient participation in decision making about care. AB - The purpose of this study was to find out how cancer patients perceive patient participation in decision-making and to see which factors in their view facilitate and restrict participation. Data were collected in focus group interviews with 25 patients, most of whom had breast cancer. Data interpretation was based on the method of qualitative content analysis. The results showed that patients, nurses and physicians all play a part in terms of how patients participate in decision-making. Patients defined participation in decision-making in terms of asking questions, obtaining/providing information and choosing from/presenting different alternatives. Among the factors that were thought to promote participation in decision-making were the patient's activity, the presence of a primary nurse/physician, the encouragement of nurses and physicians to participate, the treatment of patients as equals, and nurses and physicians having enough time for patients. As for factors hindering participation in decision-making, reference was made to patient ignorance, physical and mental imbalance and shyness on the part of the patient. Obstacles to participating in decision-making that originated in the nurses and physicians were the tendency for them to treat patients as objects, to fall in a routine, problems with information dissemination and lack of time. PMID- 11409061 TI - A meaningful relief from suffering experiences of massage in cancer care. AB - The experience of massage in an oncology ward was the focus of this study. Eight female cancer patients were given massage for 10 consecutive days and then interviewed using phenomenology as a theoretical framework. The essential meaning of getting massage as part of the daily care for female cancer patients was described as getting a meaningful relief from suffering. The findings identified five themes: the relief is meaningful because it offers the patient an experience of being "special." The massage contributes to the development of a positive relationship with the personnel, to feeling strong, and to a balance between autonomy and dependence. The massage also brings about a meaningful relief from suffering because it just "feels good." The findings of this study can be of use to health care professionals as it shows that the relatively short period of massage can result in physical and emotional benefits for cancer patients. PMID- 11409062 TI - Teaching senior nurses how to teach communication skills in oncology. AB - Many nurses acknowledge that their nursing practice is hampered by inadequate teaching about communication skills during their nursing education. Ineffective communication has negative effects on patient care and causes stress when nurses interact with each other, with medical colleagues, with patients and their relatives. Many senior nurses teach junior staff about communication and feel uncertain about their competence to do so despite recognition of its importance. This article reports data from a training initiative endorsed by the Royal College of Nursing in the United Kingdom (UK) aimed at helping senior nurses to identify their personal strengths and weaknesses when communicating, learn new teaching methods and encourage new teaching initiatives. A residential 2-day course, based on learner-centered methods was employed. One hundred and twenty nine nurses from UK cancer centers worked on personal communication problems via group discussions, video demonstrations, small group teaching exercises and role plays with professional actors. Post-course, participants reported significantly greater confidence in handling 14 common communication problem areas in cancer (p < .0001) and in 8 different areas of teaching. Participants were very enthusiastic about the course overall and especially valued the training approach and teaching materials provided. Three months post-course 91% reported changing their own teaching practice and 85% had initiated new communication skills teaching. PMID- 11409063 TI - The influence of nurses' knowledge, attitudes, and health beliefs on their safe behavior with cytotoxic drugs in Israel. AB - The purpose of this study was to examine the influence of the nurses' knowledge, attitudes, and health beliefs on their behavior and their actual usage of safety measures while handling cytotoxic drugs in their daily work surroundings. The Health Belief Model (HBM) and its extensive form, the Protection Motivation Theory (PMT), were used as the theoretical frameworks. Sixty-one nurses participated in the study, 31 hospital-based nurses daily exposed to cytotoxic drugs for the last 5 years, and 30 non-exposed community nurses. An occupational questionnaire was used to test the nurses' actual safe behavior and compliance with the recommended guidelines. A randomly selected group of exposed nurses were observed to validate their compliant behavior. A gap was found between the nurses' knowledge and their actual behavior concerning the potential risks of cytotoxic drugs and their use of protective measures (p < .005). Significant correlations were found among the components of the extensive HBM (perceived susceptibility, barriers, benefits and self-efficacy). The observational findings supported the above results. The study's findings support the need to promote primary prevention by providing a safe environment for the employee by means of education, training with regard to safety measures, clear policy, written guidelines and their enforcement. PMID- 11409064 TI - The menopausal symptom experience in young mid-life women with breast cancer. AB - Menopausal symptoms are important concerns for breast cancer survivors, which may influence daily activities, physical comfort and sexual health. Incidence and severity ratings of menopausal symptoms contribute to our knowledge about menopause in women with cancer, but fail to fully describe the symptom experience. The purpose of this article is to broaden our understanding by describing variation in menopausal symptom distress and how women interpret and manage symptoms within the context of breast cancer. From a larger grounded theory study that explored women's responses to the experience of premature induced menopause within the context of breast cancer, the constant comparative method of analysis was used to generate a detailed contextually grounded description of the menopausal symptom experience in a sample of 27 women with breast cancer who received adjuvant therapy. Women identified a symptom profile of menstrual cycle changes, hot flashes, insomnia, vaginal dryness, dyspareunia, alterations in mood, cognition and libido, and weight gain. The majority of women reported menopausal symptoms but some women were distress free while others reported moderate to severe distress. The context of breast cancer influenced women's response to symptoms and their decision making about menopausal symptom management. PMID- 11409065 TI - Perceived social support, uncertainty, and quality of life of younger breast cancer survivors. AB - This study investigated the relationship between perceived social support, uncertainty, and quality of life among younger breast cancer survivors. Proposed hypotheses predicted a significant positive correlation between perceived social support and quality of life, a significant negative correlation between uncertainty and quality of life, and that perceived social support and uncertainty, considered together, will explain more of the variance of quality of life than either variable considered independently. A sample of 101 breast cancer survivors below age 50 completed the Social Support Questionnaire, Mishel Uncertainty in Illness Scale--Community Form, and Ferrans and Powers Quality of Life Index--Cancer Version. Data were analyzed using Pearson product-moment correlation and stepwise multiple regression. All three hypotheses were supported. Additional findings revealed a significant positive correlation between support and network size, and between network size and the socioeconomic domain of quality of life. Significant negative correlations were found between perceived social support and uncertainty, between network size and time since diagnosis and treatment, and between network size and uncertainty. Study results demonstrated that although perceived social support and uncertainty accounted for a significant amount (27.2%) of variance of quality of life, a large amount of variance remains unexplained. Further research is needed in this area. PMID- 11409066 TI - Living with newly diagnosed breast cancer--the meaning of existential issues. A qualitative study of 10 women with newly diagnosed breast cancer, based on grounded theory. AB - This study aimed to describe how 10 Norwegian women diagnosed with breast cancer experienced living with the disease. A qualitative method was used, including open-ended in-depth interviews based on principles in Grounded Theory. Data revealed that existential awareness was a central phenomenon in the women's experience. This central finding created the basis for the core category in data: the will to live. This core category includes existential aspects such as different levels of life expectations, the fight against death, life related to the future, religious beliefs and doubts, and increased awareness of values in life. Knowledge and an understanding of how women experience being diagnosed with cancer are prerequisites for supporting the women in a process of normalization. This study has shown that the existential aspects connected with the core category, the will to live, are a central issue in recovery and survival. The study suggests that health professionals, by increasing their awareness of existential aspects connected with the will to live, can assist women and their families in developing coping strategies. PMID- 11409067 TI - Conventional or adhesive external breast prosthesis? A prospective study of the patients' preference after mastectomy. AB - After having been subjected to radical mastectomy for breast cancer, women are usually fitted with an external breast prosthesis. Different types of prostheses are available, but oncology nurses have few data enabling them to advise their patients adequately. In this prospective randomized crossover study of 101 women undergoing one-sided mastectomy for breast cancer, the self-adhesive breast prosthesis was compared with the traditional external prosthesis. Questionnaires were used to evaluate the woman's judgment of the prosthesis in relation to her final preference for one type of prosthesis. Complete data were available for 91 women, 59.3% of whom finally preferred the self-adhesive type. Preference was independent of age, randomization, order, or the possible use of adjuvant chemotherapy. Satisfaction with the self-adhesive prosthesis was independent of randomization order but satisfaction with the traditional type was significantly more when it was the first type of prosthesis. Preference for the self-adhesive prosthesis over the traditional type mas mainly related to an increased perception of the prosthesis as a part of the body. Preference for the traditional prosthesis over the other type was mainly related to the greater ease of application and the lesser local irritation of the skin. Findings from this study can be useful in oncology nursing practice. PMID- 11409068 TI - Partner understanding of the breast and prostate cancer experience. AB - PURPOSE: This exploratory study was designed to examine partners' understanding of patients' experience with prostate or breast cancer and to assess the congruence between patient and partner perceptions regarding social support and the cancer experience. METHODS: Partner understanding of patient mood was assessed by comparing the Profile of Mood States (POMS) questionnaire scores of patients with the POMS scores of their partners (who were instructed to complete the questionnaire as patient proxies). A semistructured interview with corresponding questions for patients and partners assessed the congruency between patient and partner perceptions regarding social support and the cancer experience. RESULTS AND CONCLUSIONS: Twenty-three couples participated. Eight couples were woman with breast cancer and their husbands, and 15 couples were men with prostate cancer and their wives. The patients with breast cancer and their partners were an average age of 48 years and had been married for 20 years, while the patients with prostate cancer and their partners were 67 years of age and had been married for 40 years, on average. Overall, female partners possessed a more accurate understanding of their husbands' experience with prostate cancer than male partners had of women's breast cancer experience. These men tended to overestimate the breast cancer patients' self-reported levels of distress. Patient and partner perceptions regarding social support in the relationship and the cancer experience were also more congruent in prostate couples than in breast couples. This may be a factor not only of gender but also of age and the length of time that these couples had been together, which was twice as long for the prostate couples. Overall, however, patients were well adjusted and felt understood by their partners and satisfied with the support their partners provided. PMID- 11409069 TI - [Inhibition of renin-angiotensin-aldosterone system: what evidence?]. AB - The activation of various neuro-humoral systems is involved in the progression of chronic heart failure. The pharmacological blockade of the renin-angiotensin aldosterone system is possible with agents such as angiotensin converting enzyme inhibitors (ACEI), the angiotensin II receptor antagonists (ARA) and the aldosterone antagonists. The actual scientific evidence supports the use of the ACEI in patient with ventricular dysfunction in all functional NYHA classes, of the ARA in heart failure patients with intolerance to ACEI and of low-dose spironolactone in patients with severe heart failure. Ongoing clinical trials will further clarify the role of these pharmacological agents in the management of chronic heart failure patients. PMID- 11409070 TI - [Adrenergic beta inhibitors in heart insufficiency: which and when?]. AB - After briefly reviewing the alterations of sympathetic nervous system in congestive heart failure and the possible ways of its pharmacological management, we review the present data regarding the use of blocking drugs in major trials (USCP, CIBIS and MERIT-HF). beta blocking drugs have a top and unquestionable role in the therapy of congestive heart failure; we conclude with a few practical rules about their use in this syndrome. PMID- 11409071 TI - [Prevention of complications: a forgotten aspect of therapy? Thromboembolic complications]. AB - In Heart Failure (HF) there is a prothrombotic state or hypercoagulation by changing of the blood flow, and by endothelial vessels or blood viscosity changes. Although these alterations in HF, the indication for an anticoagulant therapy is not precise, generally being done an empirical therapy, being still as a forgotten aspect in preventing the complications of this syndrome. When anticoagulant therapy is implemented the hemorrhagic complications risk must be carefully observed, for a significant quantity of patients may need hospital care. In spite of the existence of guidelines of the European Society of Cardiology and of the American Society of Heart Failure the decision of hypocoagulation treatment must be studied case by case, considering multiple facts, as the social state of the patient, because this therapy needs a good follow-up, a clinical surveillance and a frequent laboratory control. PMID- 11409072 TI - [Prevention of complications: a forgotten aspect of therapy? arrhythmia complication in heart failure]. AB - Heart failure has a high prevalence in the western world: 1-1.5% in the general population and 2-5% in patients above 65 years old. The association with cardiac dysarrhythmia is very common: 20-30% of patients have atrial fibrillation and significant ventricular arrhythmias occur in 50%. The pathophysiology of cardiac arrhythmias is multifactorial, including reentry, increased automaticity and triggered activity. Several prognostic factors have been described in patients with coronary artery disease, however, the two more important are the functional class and the severity of left ventricular dysfunction. In order to prevent disarrhythmic complications, the appropriate treatment of heart failure and of the underlying cardiac disease are very important. Class III antiarrhythmic drugs and the implantable cardioverter-defibrillator are increasingly used as primary (in high risk patients) or secondary prophylactic agents. PMID- 11409073 TI - [Options in drug combinations]. AB - After recalling the fundamental importance of ACE inhibitors in the treatment of heart failure, the author analyzes the scientific evidence supporting the use of beta-adrenergic blockers in the treatment of this syndrome. He describes the complications involved in prescribing these drugs for patients in functional class IV and reviews the current literature on the problem. He then considers the possibility of beta-blockers (particularly carvedilol) being used instead of ACE inhibitors. He reviews the conclusions of studies on the benefits of adding an AT1 receptor antagonist to ACE inhibitor therapy, including the results of the RESOLVD studies. He also concludes that there is no evidence that AT2 receptor antagonists are as good as or better than ACE inhibitors in the treatment of heart failure. PMID- 11409074 TI - [Surgical treatment: in which patients and what type of intervention?]. AB - In first place was described the treatment of myocardial revascularization, its indications, contra-indications and the advances in the last decade with best results. After was described the cardiomyoplasty and its results, the long-term mechanical circulatory support with various types of devices and finally the ventricular reduction (Batista op) with the short term results. PMID- 11409075 TI - [Pre-transplant evaluation: which patients for the active list?]. AB - Cardiac transplantation has been shown to significantly improve the survival and quality of life of selected patients with advanced heart failure. The shortage of appropriate donor hearts, the costs of cardiac transplantation and its associated morbidity, requiring long-term medical follow-up, mandates the judicious application of cardiac transplantation to appropriate recipients. A review of current indications, contraindications and initial evaluation of patients for cardiac transplantation is presented. PMID- 11409076 TI - [Morbidity and mortality after heart transplanation]. AB - AIM: To evaluate the morbidity and mortality after heart transplantation in the patients treated at Santa Cruz Hospital. METHODS: Between February 1986 and December 2000, 113 patients underwent orthotopic heart transplantation at Santa Cruz Hospital. Actuarial survival rates at 12 months, 5 and 10 years after surgery were calculated (Kaplan-Meier analysis). We identified the causes of death and evaluated their relation with time after transplantation. Quality of life after surgery was assessed by functional status and employment status. Main causes of morbidity were identified: infection, rejection, tumors, allograft vasculopathy and side effects of chronic immunosuppression. RESULTS: Eighty-one patients (71.7%) were discharged alive after transplantation. In this group, actuarial survival was 82.2% at 12 months, 64.6% at 5 years and 45.3% at 10 years. Maximal survival was 14.7 years. In the immediate postoperative period, the main causes of mortality were procedure-related ou primary cardiac complications. Infection was the most common cause of death in the first 3 months after transplantation. Allograft vasculopathy was the main cause of death in the group of patients surviving the first year. Malignancy was another cause of late mortality. Clinically relevant non-fatal infections occurred in 35.4% of our patients. Overall, 108 mild rejection episodes were diagnosed in 55 patients, 82 moderate rejection episodes in 52 patients and 9 severe rejections in 8 patients. Eleven non-fatal tumors were detected in 10 patients. In the group of patients who have underwent at least one coronary arteriography, 34.6% present allograft vasculopathy. Clinically relevant complications of chronic immunosuppression were: hypertension in 50% patients; dyslipidemia in 31%; osteoporosis in 19.5%; diabetes mellitus in 15%; end-stage renal failure in 3.5%. CONCLUSION: Cardiac transplantation is a valid treatment for end-stage heart failure. Most patients surviving long-term are in good physical status and their quality of life is comparable to that of the general population. PMID- 11409077 TI - [Heart insufficiency: reflecting about the present . . . preparing for the future]. PMID- 11409078 TI - [Dynamics of heart transplantation lists and anticipated needs]. AB - No other form of therapy, whether medical or surgical, has an impact comparable to heart transplantation on quality of life and survival of the patients with severe heart failure. In the EC and in the USA about 10 patients per million people are transplanted each year. Since Portugal has a population of 10 million, 100 patients should be transplanted each year. However only 8 patients were in fact transplanted in Portugal in 1998, 2 of them at Hospital S. Joao. We transplanted less than 10% of the predicted needs. We have the second lowest rate of heart transplantation per million people of the 23 European countries with registries of this activity. This is due to structural and organizational deficiencies which, at least in the case of Hospital S. Joao have a triple nature: the lack of a professional team devoted to severe heart failure treatment and heart transplantation; the lack of infrastructures, namely a heart failure ward and a heart failure outpatient clinic; and thirdly an inappropriate distribution of the tasks associated with heart transplantation and patient follow-up. A prediction of the resources needed for a successful heart transplantation program at Hospital S. Joao is presented. This was based on what is known regarding the natural history, morbidity and mortality of severe heart failure, on our own experience in this field, and finally on the experience of a large Spanish heart transplantation centre. The recommendations of the British Transplantation Society and those of the Department of Cardiothoracic Surgery of Stanford were also taken into account. PMID- 11409079 TI - [Heart transplantation. Difficulties and perspectives]. AB - In this article a revision of the several problems of cardiac transplantation in the Cardiothoracic Surgery Center of the Hospital of S. Joao is done. After a revision of the exams that have to be done and the contra-indications to surgery, the problems concerning the donors are considered. The principal conclusion to take out of is that, in the majority of the cases, the problem of the few number of cardiac transplants done, is not the scarcity of donors. PMID- 11409080 TI - [Difficulties in multi-organ harvesting]. AB - Planning in transplantation is always a difficult process mainly due to the difficulty presented in attaining the required estimated, as well as the problems inherent in transplant units. According to data from European Countries and USA, the ideal number of donors is 50/1 million inhabitants per year, 90% of whom will be multiorgan donors. In 1999 Portugal climbed to 19.2 donors/1 million inhabitants per year, 80 of whom were kidney and liver donors. A data comparison with European Countries, particularly France shows that Portugal is well placed in terms of the number of transplants units per million of population (p.m.p.). In fact, the problem lies in the area of heart and lung transplantation where the annual figures are well below requirements. Estimates for the year 2000 forecast 18 heart transplantation, % the 1999 figure. PMID- 11409081 TI - The seductive lure of creative compensation. AB - In today's booming economy, the experienced development professional is being asked to take a more active role in the management of the organization. Not only is the development professional required to raise money in the traditional sense, but she is also being asked to create programs that will raise money in cyberspace, develop e-commerce ventures and product lines. PMID- 11409082 TI - Corporations and non-profits partner to help seniors & their working caregivers. AB - Building relationships provides significant human benefits and a solid return on investment. PMID- 11409083 TI - Corporate sponsorships: increasing your slice of the pie. AB - Organizations that sell sponsorship "properties" need to understand that when corporate donors give generously to causes, they do so with specific marketing objectives in mind. PMID- 11409084 TI - Writing columns for the local media. PMID- 11409085 TI - Non-profits discover the benefits of using software through the Internet. AB - Many non-profits are looking at ways that they can leverage the Internet to assist in fund raising. Some organizations are using the Internet as a form of e commerce to accept online gifts, while others are using the immediacy of the Internet for online auctions. PMID- 11409086 TI - Achieving more for your mission. AB - Becoming more sophisticated about financial management may require outside assistance. Finding a trusted advisor who can offer expert financial advice and the tools that non-profits need to achieve financial stability and growth can empower non-profits to achieve more for their mission. PMID- 11409087 TI - Characteristics of an effective fund-raising letter. AB - When you set out to write a fund-raising letter, make sure you know precisely to whom you're writing and why--and be certain your letter makes that point just as clear to them as it is to you. PMID- 11409088 TI - The future of planned giving: seniors and the online revolution. PMID- 11409089 TI - Physicians must help make key decisions. PMID- 11409090 TI - Consumers use Internet when sick, and they favor disease-specific Web sites. AB - Hospitals seeking to use their Web sites to build and maintain relationships with patients and their families will get the best results by providing in-depth, illness-specific content instead of news and wellness information. PMID- 11409091 TI - Drug cost increases are generated by the 50 top selling pharmaceuticals. AB - Hospitals, integrated health care delivery networks (IDNs), managed care organizations, employers and the government are struggling with the soaring cost of prescription pharmaceuticals, and they will find that the 50 top selling prescription drugs are blowing their budgets. PMID- 11409092 TI - Weaker HMOs will seek concessions from hospitals for 2002 as medical cost rise. AB - The worsening financial conditions of even some of the strongest health maintenance organizations will force hospitals and integrated health care delivery networks to be even more vigilant in negotiating managed care contracts and managing their labor costs. PMID- 11409093 TI - Aetna, Cigna, report sharply higher medical costs for first quarter; say outlook is grim. AB - Health maintenance organizations' financial reports will receive close scrutiny from hospitals and physicians. PMID- 11409094 TI - HMOs are reduced to being prepaid indemnity health insurers. AB - When you go into negotiations with a managed care organization, you need to understand the economic and financial forces that are at work on HMOs and PPOs as well as the financial condition of your organization. PMID- 11409095 TI - Rural health markets need restructuring. PMID- 11409096 TI - Individual differences in working memory: introduction to the special section. AB - This special section includes a set of 5 articles that examine the nature of inter- and intraindividual differences in working memory, using working memory span tasks as the main research tools. These span tasks are different from traditional short-term memory spans (e.g., digit or word span) in that they require participants to maintain some target memory items (e.g., words) while simultaneously performing some other tasks (e.g., reading sentences). In this introduction, a brief discussion of these working memory span tasks and their characteristics is provided first. This is followed by an overview of 2 major theoretical issues that are addressed by the subsequent articles--(a) the factors influencing the inter- and intraindividual differences in working memory performance and (b) the domain generality versus domain specificity of working memory--and also of some important issues that must be kept in mind when readers try to evaluate the claims regarding these 2 theoretical issues. PMID- 11409097 TI - A controlled-attention view of working-memory capacity. AB - In 2 experiments the authors examined whether individual differences in working memory (WM) capacity are related to attentional control. Experiment 1 tested high and low-WM-span (high-span and low-span) participants in a prosaccade task, in which a visual cue appeared in the same location as a subsequent to-be-identified target letter, and in an antisaccade task, in which a target appeared opposite the cued location. Span groups identified targets equally well in the prosaccade task, reflecting equivalence in automatic orienting. However, low-span participants were slower and less accurate than high-span participants in the antisaccade task, reflecting differences in attentional control. Experiment 2 measured eye movements across a long antisaccade session. Low-span participants made slower and more erroneous saccades than did high-span participants. In both experiments, low-span participants performed poorly when task switching from antisaccade to prosaccade blocks. The findings support a controlled-attention view of WM capacity. PMID- 11409098 TI - What limits children's working memory span? Theoretical accounts and applications for scholastic development. AB - Explanations of working memory span in children were studied in a longitudinal follow-up of J. N. Towse, G. J. Hitch, and U. Hutton (1998). Reading span and operation span were lower when within-task retention intervals were lengthened. For each task, variation in span between test waves and age cohorts was systematically related to changes in processing speed. The two spans explained substantial shared variance in both reading and arithmetic scores, with some evidence for domain specificity. Combined span scores predicted unique variance in scholastic attainment over a 1-year interval. The authors concluded that working memory span is constrained by rapid loss of active codes and is not simply a measure of capacity for resource sharing. Working memory is also implicated in scholastic development. PMID- 11409099 TI - Working memory span and the role of proactive interference. AB - The authors investigated the possibility that working memory span tasks are influenced by interference and that interference contributes to the correlation between span and other measures. Younger and older adults received the span task either in the standard format or one designed to reduce the impact of interference with no impact on capacity demands. Participants then read and recalled a short prose passage. Reducing the amount of interference in the span task raised span scores, replicating previous results (C. P. May, L. Hasher, & M. J. Kane, 1999). The same interference-reducing manipulations that raised span substantially altered the relation between span and prose recall. These results suggest that span is influenced by interference, that age differences in span may be due to differences in the ability to overcome interference rather than to differences in capacity, and that interference plays an important role in the relation between span and other tasks. PMID- 11409100 TI - Using working memory theory to investigate the construct validity of multiple choice reading comprehension tests such as the SAT. AB - When taking multiple-choice tests of reading comprehension such as the Scholastic Assessment Test (SAT), test takers use a range of strategies that vary in the extent to which they emphasize reading the questions versus reading the passages. Researchers have challenged the construct validity of these tests because test takers can achieve better-than-chance performance even if they do not read the passages at all. By using an individual-differences approach that compares the relative power of working memory span to predict SAT performance for different test-taking strategies, the authors show that the SAT appears to be tapping reading comprehension processes as long as test takers engage in at least some reading of the passages themselves. PMID- 11409101 TI - The relationships among working memory, math anxiety, and performance. AB - Individuals with high math anxiety demonstrated smaller working memory spans, especially when assessed with a computation-based span task. This reduced working memory capacity led to a pronounced increase in reaction time and errors when mental addition was performed concurrently with a memory load task. The effects of the reduction also generalized to a working memory-intensive transformation task. Overall, the results demonstrated that an individual difference variable, math anxiety, affects on-line performance in math-related tasks and that this effect is a transitory disruption of working memory. The authors consider a possible mechanism underlying this effect--disruption of central executive processes--and suggest that individual difference variables like math anxiety deserve greater empirical attention, especially on assessments of working memory capacity and functioning. PMID- 11409102 TI - The effect of distinctive parts on recognition of depth-rotated objects by pigeons (Columba livia) and humans. AB - To explore whether effects observed in human object recognition represent fundamental properties of visual perception that are general across species, the authors trained pigeons (Columba livia) and humans to discriminate between pictures of 3-dimensional objects that differed in shape. Novel pictures of the depth-rotated objects were then tested for recognition. Across conditions, the object pairs contained either 0, 1, 3, or 5 distinctive parts. Pigeons showed viewpoint dependence in all object-part conditions, and their performance declined systematically with degree of rotation from the nearest training view. Humans showed viewpoint invariance for novel rotations between the training views but viewpoint dependence for novel rotations outside the training views. For humans, but not pigeons, viewpoint dependence was weakest in the 1-part condition. The authors discuss the results in terms of structural and multiple view models of object recognition. PMID- 11409103 TI - Timing and reaction time. AB - Because reaction time (RT) tasks are generally repetitive and temporally regular, participants may use timing strategies that affect response speed and accuracy. This hypothesis was tested in 3 serial choice RT experiments in which participants were presented with stimuli that sometimes arrived earlier or later than normal. RTs increased and errors decreased when stimuli came earlier than normal, and RTs decreased and errors increased when stimuli came later than normal. The results were consistent with an elaboration of R. Ratcliff's diffusion model (R. Ratcliff, 1978; R. Ratcliff & J. N. Rouder, 1998; R. Ratcliff, T. Van Zandt, & G. McKoon, 1999), supplemented by a hypothesis developed by D. Laming (1979a, 1979b), according to which participants initiate stimulus sampling before the onset of the stimulus at a time governed by an internal timekeeper. The success of this model suggests that timing is used in the service of decision making. PMID- 11409104 TI - Grounding spatial language in perception: an empirical and computational investigation. AB - The present paper grounds the linguistic cdategorization of space in aspects of visual perception; specifically, the structure of projective spatial terms such as above are grounded in the process of attention and in vector-sum coding of overall direction. This is formalized in the attentional vector-sum (AVS) model. This computational model accurately predicts linguistic acceptability judgments for spatial terms, under a variety of spatial configurations. In 7 experiments, the predictions of the AVS model are tested against those of 3 competing models. The results support the AVS model and disconfirm its competitors. The authors conclude that the structure of linguistic spatial categories can be partially explained in terms of independently motivated perceptual processes. PMID- 11409105 TI - Cognitive arithmetic across cultures. AB - Canadian university students either of Chinese origin (CC) or non-Asian origin (NAC) and Chinese university students educated in Asia (AC) solved simple arithmetic problems in the 4 basic operations (e.g., 3 + 4, 7 - 3, 3 x 4, 12 divided by 3) and reported their solution strategies. They also completed a standardized test of more complex multistep arithmetic. For complex arithmetic, ACs outperformed both CCs and NACs. For simple arithmetic, however, ACs and CCs were equal and both performed better than NACs. The superior simple-arithmetic skills of CCs relative to NACs implies that extracurricular culture-specific factors rather than differences in formal education explain the simple-arithmetic advantage for Chinese relative to non-Asian North American adults. NAC's relatively poor simple-arithmetic performance resulted both from less efficient retrieval skills and greater use of procedural strategies. Nonetheless, all 3 groups reported using procedures for the larger simple subtraction and division problems, confirming the importance of procedural knowledge in skilled adults' performance of elementary mathematics. PMID- 11409106 TI - Why stem cells will transform medicine. PMID- 11409107 TI - Biotechnology. Bubble, bubble, toil and trouble. PMID- 11409108 TI - Treatment of rheumatoid arthritis 2001. PMID- 11409109 TI - The genetics of cytokines in ankylosing spondylitis. PMID- 11409110 TI - When, why, and how should we quantify the excretion rate of urinary uric acid? PMID- 11409111 TI - Serum cytokines in different histological variants of rheumatoid arthritis. AB - OBJECTIVE: Rheumatoid arthritis (RA) is characterized by an invasive and tissue destructive infiltrate of lymphocytes, macrophages, and synoviocytes formed in the joints. Its etiopathogenesis and the role of the particular morphological components of synovitis remain unclear. There is evidence that its histological heterogeneity is correlated with synovium cytokine transcription. We investigated whether the serum cytokine profile is associated with the morphological appearance of the disease. METHODS: Tissue and serum samples were collected from 25 patients with clinically active RA and 25 with osteoarthritis (OA) as a control group. After histological analysis RA synovial biopsies were divided into 2 distinct types; 16 samples were characterized by diffuse lymphocyte infiltrates with no additional microanatomical organization. Lymphocytic aggregates with germinal center-like structures were found in 9 specimens. Serum concentrations of interferon-gamma (IFN-gamma), interleukin 12 (IL-12, p70 heterodimer), tumor necrosis factor-alpha (TNF-alpha), and IL-15 were measured by ELISA. RESULTS: Low concentrations of IFN-gamma (p < 0.01) and IL-12 (NS) were found in RA patients' serum compared with OA controls. RA patients with follicular synovitis had lower serum concentration of IFN-gamma (p < 0.05) and IL-12 (p < 0.05) than patients with diffuse infiltrates. High concentration of TNF-alpha and IL-15 characterized RA patient serum in comparison with controls (respectively, p < 0.001 and p < 0.01). In the serum of RA patients with follicular synovitis TNF-alpha was a dominant cytokine (p < 0.01) compared to patients with diffuse disease. At TNF alpha level > or = 44 pg/ml, 5 (56%) of 9 patients with follicular RA had such elevated values vs one of 16 diffuse patients (< 10%; p < 0.02). Only serum concentrations of TNF-alpha could effectively differentiate between patients with OA and subgroups of RA. Analysis of clinical data suggested that activity of rheumatoid disease in patients with follicular synovitis was more severe than in those with diffuse infiltrates. CONCLUSION: The association between distinct histological appearance of rheumatoid synovitis and serum cytokine profile and diverse clinical activity of disease seems to confirm its heterogeneity. PMID- 11409112 TI - Rheumatologists' attitudes toward routine screening for hydroxychloroquine retinopathy. AB - OBJECTIVE: To examine rheumatologists' attitudes toward routine ophthalmologic screening for hydroxychloroquine (HCQ) retinopathy and to estimate the acceptability of hypothetical guidelines discouraging this practice. METHODS: We E-mailed a random sample of 300 US rheumatologists listed in the American College of Rheumatology (ACR) Directory who treat adults. We asked about current screening practice, reasons for screening, and the effect of hypothetical guidelines discouraging routine screening on future practice. Associations between adherence to guidelines and clinical variables were evaluated using multiple logistic regression. RESULTS: Of 56% who responded, almost all (94%) currently screen their patients at least once per year. Seventy-five percent stated that they would continue to screen because they are unwilling to accept any risk of visual loss among their patients; 74% would continue to screen because of legal liability; and 56% felt their patients would insist on being screened regardless of their physician's opinion. Forty-four percent stated that they would continue to screen regularly, even if the ACR published guidelines discouraging routine screening. Rheumatologists unwilling to accept any risk of retinopathy were less likely to follow guidelines discouraging screening (46 vs 77%, adjusted OR 0.2, 95% CI 0.1-0.6). Patient insistence and fear of legal liability were not significantly associated with predicted adherence to guidelines. CONCLUSION: Our survey indicates that the majority of rheumatologists currently routinely screen their patients for HCQ retinopathy, and that many would not follow ACR guidelines discouraging this practice, at least in part because they are unwilling to accept any risk of visual damage. PMID- 11409113 TI - Mechanisms of CD23 hyperexpression on B cells from patients with rheumatoid arthritis. AB - OBJECTIVE: To analyze the mechanisms involved in the characteristic hyperexpression of CD23 on peripheral blood B cells from patients with rheumatoid arthritis (RA). METHODS: Peripheral blood mononuclear cells (PBMC) were obtained from patients with active disease and activated during 18 h with an anti-CD3 monoclonal antibody in the presence or absence of blocking antibodies to CD154 or CD40. PBMC were further purified by rosetting and CD23 expression was assessed on B cells by flow cytometry after double staining (CD19/CD23). Lymphocytes were also isolated from synovial fluid (SF). CD154 expression was analyzed on PB or SF CD4+ T cells after double staining (CD4/CD154) by flow cytometry at basal conditions and after different stimuli [anti-CD3 or phorbol myristic acetate (PMA) plus ionomycin]. Co-culture experiments between SF and PB cells were performed to analyze the involvement of the CD40-CD154 interaction on CD23 expression. CD154 and CD23 expression was also analyzed on synovial membrane by immunohistochemical techniques. RESULTS: A high proportion of activated CD23 B cells was detected in patients with RA. Blocking experiments with both anti-CD40 and anti-CD154 Mab showed a significant reduction in the proportion of PB B cells expressing CD23. Following activation with anti-CD3 Mab or PMA plus ionomycin, CD154 expression was mainly induced on PB CD4+ T cells. In co-culture experiments, SF T cells were more efficient than PB T cells in inducing CD40 dependent CD23 expression on PB B cells. In addition, CD4+ T cells from synovial membrane clearly expressed CD154. CONCLUSION: Our results establish a link between CD154-CD40 pathway and CD23 expression on PB B cells from patients with RA. T cells from the synovial microenvironment were active participants in this CD23 expression, presumably in the context of cell recirculation. PMID- 11409114 TI - KE-298 and its active metabolite KE-758 suppress nitric oxide production by murine macrophage cells and peritoneal cells from rats with adjuvant induced arthritis. AB - OBJECTIVE: To analyze the effects of KE-298 and KE-758 on lipopolysaccharide (LPS) induced nitric oxide (NO) production by the RAW264.7 murine macrophage cell line, and the effect of KE-758 on spontaneous NO production by peritoneal cells from rats with adjuvant induced arthritis. METHODS: The amount of NO was determined using Griess reagents. The proteins for inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) were detected by Western blot, then mRNA for interferon-beta (IFN)-beta, IFN regulatory factor-1 (IRF-1), and iNOS were detected by RT-PCR. Degradation of iNOS mRNA was analyzed using Northern blot. Nuclear factor-kappa B (NF-kappa B) in nuclear extracts was determined by EMSA. Adjuvant arthritis in rats was induced by inoculating heat killed Mycobacterium butyricum s.c. in the tail. RESULTS: KE-298 and KE-758 suppressed NO production by LPS activated RAW264.7 cells by inhibiting iNOS gene expression. Neither LPS induced NF-kappa B activation nor degradation of iNOS mRNA was affected by KE-758 treatment. LPS induced IFN-beta and IRF-1 gene expression were markedly suppressed by KE-758. In rats with adjuvant induced arthritis, enhanced NO and iNOS production by cultured peritoneal cells and the development of arthritis were suppressed by KE-758. CONCLUSION: KE-758 suppressed LPS induced iNOS gene expression by murine macrophage cells by inhibiting IFN-beta/IRF-1 expression. The potential of KE-758 to inhibit iNOS production might partly explain its efficacy on adjuvant induced arthritis in rats. PMID- 11409115 TI - Long-term safety and efficacy of etanercept in patients with rheumatoid arthritis. AB - OBJECTIVE: Patients with rheumatoid arthritis (RA) treated with etanercept (Enbrel) in controlled studies of 3 to 6 months' duration had rapid and sustained improvement of their disease, with minimal safety issues. In this study, we examine safety and clinical benefit after longer term treatment with etanercept. METHODS: All adult patients with RA with a previously inadequate response to one or more disease modifying antirheumatic drugs, and who received at least one dose of etanercept as monotherapy in controlled or open label clinical trials were evaluated for safety and clinical benefit. Adverse event rates were compared as was evidence of continued benefit over time. RESULTS: Etanercept continued to be safe and well tolerated in 628 adult patients treated for a median of 25 mo (maximum 43 mo; 1109 patient-years). Nine percent of patients withdrew due to lack of efficacy and 7% due to adverse events. Most adverse events were mild, and no statistically significant increases in frequency of events were seen when patients received etanercept over longer periods of time. Clinical benefit was maintained with longterm therapy. A 100% improvement in individual disease activity measures was achieved by 17% to 28% of the patients. Fifty-five percent of patients who were taking corticosteroids (mean dose at baseline 6.6 mg/day) decreased or discontinued corticosteroid therapy while maintaining control of their arthritis symptoms. CONCLUSION: Etanercept continued to be safe and well tolerated, and its clinical benefit was sustained for a median of 25 mo and for as long as 43 mo in patients with RA. PMID- 11409116 TI - Increased frequency of interleukin 4 producing CD4+ and CD8+ cells in peripheral blood from patients with systemic sclerosis. AB - OBJECTIVE: To-determine the relationship between the frequency of cytokine producing cells and systemic sclerosis (SSc), we examined the frequencies of interleukin 4 (IL-4) or IL-2 producing CD4+ or CD8+ T cells in peripheral blood mononuclear cells (PBMC) from patients with SSc. METHODS: PBMC from 23 SSc patients and 14 healthy controls were isolated from heparinized blood by density centrifugation. Purified PBMC were stimulated with immobilized anti-CD3 monoclonal antibody for 6 h in the presence of monensin. Cells were fixed, made permeable, and stained for intracellular cytokines in combination with staining for T cell surface markers, CD4, and CD8. RESULTS: The frequency of CD4+ T cells was negatively correlated with duration of SSc, whereas the frequency of CD8+ T cells was not correlated with duration. The frequencies of IL-4 producing (IL-4+) cells in CD4+ T cells and in CD8+ T cells from patients with SSc were both significantly higher than those from controls. In contrast, the frequencies of IL 2 producing (IL-2+) cells in CD4+ T cells and in CD8+ T cells from patients with SSc were both significantly lower than that from controls. Further, the ratios of IL-4+/IL-4+ + IL-2+ in CD4+ T cells and in CD8+ T cells were both negatively correlated with disease duration of SSc. CONCLUSION: These results suggest that type 2 cytokine producing T cells, not only CD4+ T cells but also CD8+ T cells, have important roles in the pathogenesis of SSc, especially in the early phase of SSc. PMID- 11409117 TI - Low serum dehydroepiandrosterone sulfate in women with primary Sjogren's syndrome as an isolated sign of impaired HPA axis function. AB - OBJECTIVE: To assess the hypothalamic-pituitary-adrenal (HPA) and thyroid axes in women with primary Sjogren's syndrome (pSS). METHODS: In 10 women with pSS and 10 age matched female controls, we evaluated serum dehydroepiandrosterone sulfate (DHEA-S), testosterone, androstenedione, follicle stimulating hormone, luteinizing hormone, thyroid stimulating hormone, prolactin, growth hormone, sex hormone binding globulin, cortisol, and adrenocorticotropin hormone (ACTH), in both basal condition and after stimulation with corticotropin releasing hormone, thyrotropin releasing hormone, and luteinizing hormone releasing hormone intravenously. Patients had not previously been treated with glucocorticoids. RESULTS: Patients with pSS had significantly lower basal mean DHEA-S values compared with healthy controls (2.4 +/- 0.4 vs 3.9 +/- 0.3 mumol/l; p < 0.05) and significantly lower DHEA-S values after stimulation. The cortisol/DHEA-S ratio in the patient group was higher than in controls (171 +/- 39 vs 76 +/- 5; p < 0.05). A correlation was found between basal ACTH and DHEA-S values in the patients (r = 0.650; p = 0.05). No correlation was seen between disease activity or age and the serum concentration of DHEA-S. The levels of other hormones both at baseline and after stimulation were similar in patients and controls. CONCLUSION: The results show that women with pSS have intact cortisol synthesis but decreased serum concentrations of DHEA-S and increased cortisol/DHEA-S ratio compared with healthy controls. The findings may reflect a constitutional or disease mediated influence on adrenal steroid synthesis. The thyroid axis and gonadotropin secretion were similar in patients and controls. PMID- 11409118 TI - HLA-DRB1*01 association with Henoch-Schonlein purpura in patients from northwest Spain. AB - OBJECTIVE: To examine the HLA-DRB1 phenotypes of patients with Henoch-Schonlein purpura (HSP) and determine if associations exist with disease susceptibility, clinical heterogeneity, or severe systemic complications. METHODS: A retrospective study was performed on an unselected population of patients from Northwest Spain with HSP classified according to proposed criteria. Patients were included in this study if they had at least one year of followup. Fifty Caucasian patients (25 women), 11 of them older than 20 years, were studied. Patients and ethnically matched controls were HLA-DRB1 genotyped from DNA using molecular based methods. RESULTS: During the course of the disease, renal manifestations, especially hematuria, and severe gastrointestinal (GI) manifestations (bowel angina or GI bleeding) were observed in more than 60% of the patients. Twenty percent of patients had persistent renal involvement (renal sequelae). Patients with HSP had a significantly higher frequency of the HLA-DRB1*01 phenotype compared to matched controls. The HLA-DRB1*07 phenotype was also significantly reduced compared with controls. Patients with severe GI manifestations or with persistent renal involvement did not exhibit any specific HLA-DRB1 association other than the underlying association with HLA-DRB1*01. CONCLUSION: HSP in a population from Northwest Spain is significantly associated with HLA-DRB1*01. PMID- 11409119 TI - Combined use of power Doppler and gray-scale sonography: a new technique for the assessment of inflammatory myopathy. AB - OBJECTIVE: Ultrasonography (US) is a utilitarian approach to the assessment of inflammatory myopathy (IM). Power Doppler sonography (PDS), a newer technique, enables detection of muscle vascularity and inflammation. We describe the combined use of PDS and gray-scale US in patients with IM. METHODS: We studied 37 IM subjects and 6 control subjects. Clinical scores of muscle strength and function were obtained. The maximum score, 31, represented normal function and strength. Ultrasonographic gray-scale and vascularity results were scored 0-4. Nine subjects had serial assessments. RESULTS: Subjects ranged from 16 to 83 years of age and were predominantly female. IM subjects had significantly abnormal lower clinical scores than controls, 23.0 +/- 5.8 vs 29.8 +/- 2.0 (p < 0.001). Mean peak gray-scale score was 2.1 +/- 0.96 compared to 0.5 +/- 0.84 for controls (p = 0.001), indicating atrophy in the IM group. Similar results were found for average gray-scale scores. Peak vascularity scores were higher in IM, 2.7 +/- 0.8 vs 2.2 +/- 0.3 (p = 0.007). Disease of longer duration was significantly associated with more abnormal gray-scale scores and lower creatine phosphokinase (CPK) levels. PDS scores were more abnormal in disease of shorter duration. There was a negative association between functional scores and inflammatory scores on serial assessment. CONCLUSION: Sonography is a valuable tool in the assessment of IM. Gray-scale and PDS findings were significantly different between IM and control subjects. Abnormal gray-scale US scores were associated with disease of longer duration and lower CPK levels. In contrast, increased vascularity on PDS detected disease of shorter duration and varied with the clinical course more than did gray-scale findings. PMID- 11409120 TI - Intercellular adhesion molecule-1 gene polymorphisms in Behcet's Disease. AB - OBJECTIVE: Intercellular adhesion molecule 1 (ICAM-1) is strongly expressed in vascular endothelial cells and perivascular inflammatory infiltrates in immunopathologic studies of Behcet's disease (BD) lesions. ICAM-1 genes may contribute to the inflammatory events responsible for the vessel damage in BD. We examined potential associations of ICAM-1 gene polymorphisms with BD susceptibility. METHODS: Case patients were 74 consecutive Italian patients with BD who were followed at the Bologna, Ferrara, Milano, Potenza, Prato, Reggio Emilia, and Trento rheumatology, ophthalmology, and neurology units over a 3 year period (1997-99) who satisfied the International Study Group criteria for BD; 228 healthy Italian blood donors from the same geographic areas were selected as control groups. All BD patients and controls were genotyped by polymerase chain reaction and allele-specific oligonucleotide techniques for ICAM-1 polymorphisms at codon 241 (exon 4) and codon 469 (exon 6). RESULTS: The frequency of R241 was significantly higher in BD patients than in controls (20.3% vs 5.7%; p = 0.001, pcorr = 0.002, OR 4.2, 95% CI 1.9-9.3). The distribution of E/K 469 genotype was similar in patients and controls. Comparing patients with different clinical features, we found only a trend to higher frequency of R241 in patients with articular manifestations (21.4% vs 12.5%; p = 0.08). CONCLUSION: Our findings show that G/R 241 polymorphism of ICAM-1 is associated with BD susceptibility. PMID- 11409121 TI - TNF-238A promoter polymorphism contributes to susceptibility to ankylosing spondylitis in HLA-B27 negative patients. AB - OBJECTIVE: To investigate the relative contribution of MHC loci in their susceptibility to primary ankylosing spondylitis (AS) in HLA-B27 negative patients and to compare the clinical features and genetic factors with those of HLA-B27 positive AS. METHODS: DNA from patients with B27 negative primary AS (n = 28), B27 positive primary AS (n = 77), and matched healthy controls (B27-, n = 100; B27+, n = 70) were analyzed to investigate whether HLA genes determine the disease susceptibility, or whether other closely linked loci might play a role in disease development. HLA typing was carried out by serology and PCR/SSP (HLA-B, DR), MICA-TM polymorphism in the transmembrane region by radioactive PCR, and tumor necrosis factor-alpha (TNF-alpha) promoter polymorphism at positions -238 and -308 by PCR-RFLP. RESULTS: Subtle clinical differences were found for primary AS, the B27 negative patients being less frequently complicated by acute anterior uveitis and more associated with peripheral arthritis than B27 positive. Differences were found in the distribution of TNF-alpha -238 genotypes among patients with primary AS (B27- vs B27+). The TNF-alpha -238(A) polymorphism was present in 50% of the B27 negative patients carrying the -238 G/A and A/A genotypes and was significantly increased compared with B27 positive AS (odds ratio 4.3) and with the B27 negative control group (OR 5.9). The TNF-alpha genotypes were equally prevalent in B27 positive AS and healthy matched B27 positive controls. No significant HLA and MICA typing differences were found between the populations under study. CONCLUSION: Our results indicate that the polymorphism variation in the TNF-alpha promoter -238.2(A) influences disease susceptibility in B27 negative primary AS but had no effect in our B27 positive AS population. PMID- 11409122 TI - Lack of association with spondyloarthritis and HLA-B27 in Italian patients with Whipple's disease. AB - OBJECTIVE: To examine consecutive patients with Whipple's disease (WD) for the full clinical spectrum of spondyloarthritis. METHODS: Nine consecutive patients, 8 men, 1 woman, with WD were clinically evaluated and examined for clinical and radiological manifestations of spondyloarthritis by 2 rheumatologists with special interest in spondyloarthritis. The mean age of the patients at the time of study and at the time of their initial diagnosis of WD by duodenal biopsy was 62.4 years (range 42-71) and 54.4 years (range 40-62), respectively. Each patient had an anteroposterior radiographic view of the pelvis and also had HLA typing for class I and II alleles. Pelvis radiographs were read blindly for evidence of sacroiliitis. RESULTS: All but one patient had rheumatologic manifestations related to WD. The mean interval between the onset of these symptoms and the diagnosis of WD was 6.1 years (range 1-15). One had recurrent polyarthritis, 1 recurrent oligoarthritis, 3 recurrent monoarthritis, 2 recurrent synovitis with pitting edema of the dorsum of hands and/or feet, and 1 isolated De Quervain's tenosynovitis. Of the 5 patients with arthritis, 3 also had episodes of swelling with pitting edema over the dorsum of hands and/or feet together with flexor digit tenosynovitis, 2 also had olecranon bursitis, and 1 tibial tenosynovitis. No patient had clinical or radiological manifestations of spondyloarthritis. None had the HLA-B27. CONCLUSION: In Italian Caucasian patients with WD there was no association with spondyloarthritis and HLA-B27. PMID- 11409123 TI - Variations in glucocorticoid induced osteoporosis prevention in a managed care cohort. AB - OBJECTIVE: To characterize glucocorticoid use and patterns of osteoporosis prevention therapies among a large US national cohort. METHODS: Health maintenance organization (HMO) members who were receiving chronic glucocorticoid therapy (> 90 day supply) within a 3 year observation period were identified along with their prescribing physicians. Receipt of anti-osteoporotic prescription therapies and bone mass measurement was determined. Multivariable analyses were used to define significant predictors of these preventive interventions. RESULTS: We identified 2378 HMO members who filled prescriptions for at least a 90 day supply of glucocorticoids, but had not filled a glucocorticoid prescription in the prior 90 days. In women over age 50, use of anti-osteoporotic therapies and bone mass measurement was 41% and 16%, respectively. Glucocorticoid-prescribing physicians were identified for 878 (37%) of these glucocorticoid users, and internal medicine specialists (39%) and rheumatologists (20%) wrote the majority of the prescriptions for glucocorticoids. Women age 50 and over were most likely to receive a prescription anti-osteoporotic preventive therapy (OR 4.0; 95% CI 1.5-10.8). Patients with a rheumatologist prescribing their glucocorticoids were more likely than those of internists to have a bone mass measurement (OR 2.2; 95% CI 1.3-3.6) and receive bisphosphonates (OR 1.9; 95% CI 1.1-3.1), but were not more likely to receive preventive treatment overall. CONCLUSION: Although better than in several prior studies, we identified low levels of selected preventive care measures for chronic glucocorticoid users in a large population based cohort. Significant demographic and practice pattern variation suggests opportunities for targeted preventive interventions. PMID- 11409124 TI - Spot urine uric acid to creatinine ratio used in the estimation of uric acid excretion in primary gout. AB - OBJECTIVE: Uric acid overexcretion in patients with gout is frequently assessed by the measurement of 24 hour urinary uric acid excretion, which is cumbersome with ambulatory patients, and requires accurate timing and complete collection of the specimen. We assessed whether uric acid to creatinine ratio (Uua/Ucr) in spot urine is useful for the estimation of uric acid overexcretion in patients with gout. METHODS: One hundred thirty male patients with gout and 33 non-gout male control subjects were studied. Early morning urine and/or a portion of 24 h collected urine (24 h urine) were used as spot urine samples. Uric acid overexcreters were defined as those with a 24 h urinary uric acid excretion > or = 1000 mg/day, while uric acid underexcreters were defined as those with uric acid clearance < 6 ml/min. RESULTS: There was a significant relationship between 24 h urinary uric acid excretion and early morning urine Uua/Ucr in patients with gout, while no such relationship was observed in controls. No significant difference in Uua/Ucr was observed between patients with gout and controls, or in Uua/Ucr between gout uric acid overexcreters and underexcreters in early morning urine. A significant difference in this value was observed between the 2 groups in the 24 h urine specimens. Although the diagnostic accuracy of gout uric acid overexcretion was 87.2% using early morning urine and 89.6% using 24 h urine, the sensitivity of gout uric acid overexcretion was only 25.0% when using early morning urine and 25.0% when using 24 h urine, when the cutoff value of Uua/Ucr was 0.63 and 0.64, respectively. CONCLUSION: Uua/Ucr using spot urine, especially early morning urine, is not an accurate indicator of uric acid overexcretion in patients with gout. PMID- 11409125 TI - Correlation of collagen organization with polarization sensitive imaging of in vitro cartilage: implications for osteoarthritis. AB - OBJECTIVE: Optical coherence tomography (OCT), a new method of high resolution imaging, has shown feasibility for assessing articular cartilage to identify early changes in osteoarthritis (OA) and monitor therapy. OCT is analogous to ultrasound, measuring the intensity of backreflected infrared light rather than sound. The resolution of this technology is up to 25 times higher than existing methods. We investigated the correlation between changes observed by OCT and the degree of collagen organization in OA cartilage. METHODS: Polarization sensitive OCT (PSOCT) imaging was used to assess changes in cartilage collagen organization in vitro. RESULTS: The presence (or absence) of PSOCT changes correlated with collagen organization (or disorganization) on histology as assessed by picrosirius polarization microscopy (no significant difference). In multiple cases, cartilage was abnormal by both PSOCT and polarization microscopy, but was grossly normal by routine staining, showing cartilage thickness > 2 mm and no fibrillations. CONCLUSION: This in vitro study suggests PSOCT changes in cartilage are due to the state of collagen organization. The combination of high resolution structural imaging and birefringence detection make OCT a potentially powerful technology for early assessment of OA. PMID- 11409126 TI - Gelatinolytic and collagenolytic activity in periprosthetic tissues from loose hip endoprostheses. AB - OBJECTIVE: To study the contribution of different members of the metalloproteinases (MMP) family in gelatinolytic and collagenolytic potential, namely dinitrophenyl-Pro-Gln-Gly-Ile-Ala-Gly-Gln-D-Arg (DNP-S) sensitive proteolytic activity, in loose total hip arthroplasty (THA) endoprostheses. METHODS: Periprosthetic tissues and fluid samples were collected from patients subjected to hip endoprosthesis replacement. DNP-S sensitive proteolytic activity was evaluated by the degradation of synthetic DNP-S and reverse phase high performance liquid chromatography, while gelatinolytic activity was assessed by gelatin zymography. The isolation and separation of gelatinases was performed by gelatin- and concanavalin A-Sepharose affinity chromatographies and the identification of collagenases by immunoblot analysis. RESULTS: High gelatinolytic activity was observed in all periprosthetic tissue extracts and fluid samples. All samples also exhibited DNP-S degrading activity, without pretreatment by activating agents. Upon fractionation of MMP by gelatin-Sepharose affinity chromatography it was found that the gelatin-unbound collagenases are exclusively responsible for DNP-S degrading activity. Activated species of both MMP-1 and 13 were detected in most samples, but not the soluble form of MT1-MMP. Separation of gelatinases from each other and treatment with 4 aminophenylmercuric acetate (APMA) revealed that both enzymes mainly existed in complex with tissue inhibitor of metalloproteinase (TIMP). CONCLUSION: MMP-1 and MMP-13, which exist in activated form, could be responsible for the DNP-S degrading activity in periprosthetic tissues and fluids, while the gelatinases do not contribute in this potential, since they mainly exist in complex with TIMP. The 2 collagenases may play a key role in the loosening of THA endoprostheses. PMID- 11409127 TI - Knee effusions, popliteal cysts, and synovial thickening: association with knee pain in osteoarthritis. AB - OBJECTIVE: To evaluate the association of effusions, popliteal cysts, and synovial thickening with knee symptoms in older persons with and without radiographic (XR) osteoarthritis (OA), using magnetic resonance imaging (MRI). METHODS: Subjects with and without knee symptoms were recruited from Veterans Affairs and community sources. All had weight-bearing knee radiographs. Subjects were divided into 3 groups: Knee pain/XROA group had knee symptoms and radiographic OA; No knee pain/XROA group had no knee symptoms and radiographic OA; and No knee pain/no XROA group had no knee symptoms and a normal radiograph. A single knee was imaged using a 1.5 T MR scanner using T1 and T2 weighted and proton density SE imaging sequences. MRI were read for effusion, popliteal cysts, and synovial thickening. RESULTS: The mean age of subjects was 67.0 years (66.6% male). We studied 381 subjects with Knee pain/XROA, 52 with No knee pain/XROA, and 25 with No knee pain/no XROA. The prevalence of moderate or larger effusions was: Knee pain/XROA 54.6%, No knee pain/XROA 15.6%, and No knee pain/no XROA 11.1%. Popliteal cysts were present in 33.0% of Knee pain/XROA subjects, 28.0% No knee pain/XROA, and 9.1% No knee pain/no XROA. After adjusting for the severity of radiographic OA, there was a difference between those with and without knee pain in prevalence of moderate or larger effusions (p < 0.001) and synovial thickening, independent of effusion (p < 0.001), but not in the prevalence of popliteal cysts. Further, among those in Knee pain/OA group, synovial thickening was associated with the severity of knee pain. CONCLUSION: Effusions and popliteal cysts are common in middle aged and elderly people. After adjusting for the degree of radiographic OA, moderate or large effusions and synovial thickening were more frequent among those with knee pain than those without pain, suggesting these features are associated with the pain of knee OA. In those with knee symptoms, synovial thickening is uniquely associated with the severity of knee pain. PMID- 11409128 TI - Sonography in the study of metatarsalgia. AB - OBJECTIVE: To identify sonographically the changes of the forefoot in patients with metatarsalgia. METHODS: Sonography of the foot was performed in 112 patients with metatarsalgia and in 50 healthy controls. Metatarsophalangeal (MTP) joints, intermetatarsal web spaces, flexor and extensor tendons, and plantar aponeurosis were examined. RESULTS: Sonography showed intermetatarsophalangeal bursitis in 20.5% of cases, Morton's neuroma in 15.2%, and effusion of MTP joints in 11.7%. CONCLUSION: Sonography gives useful information about the possible alteration responsible for metatarsalgia. PMID- 11409129 TI - Effect of intraarticular hyaluronan injection on synovial fluid hyaluronan in the early stage of canine post-traumatic osteoarthritis. AB - OBJECTIVE: To determine how the quantity and molecular weight of synovial fluid hyaluronan (HA) within the synovial fluid (SF) of osteoarthritis (OA) joints is affected by intraarticular injection of HA. METHODS: Dogs in which OA was induced by transection of the anterior cruciate ligament received 5 weekly injections of HA (1.5 x 10(6) Da) in saline (10 mg/0.67 ml) or an equal volume of saline into the operated knee, beginning the day after surgery. Immediately before each injection, SF was aspirated and the volume of SF and the concentration of HA was measured (uronic acid), and the molecular weight of the HA in each sample was estimated by electrophoresis in agarose. RESULTS: The volume of SF in the unstable knee increased after surgery, and the molecular weight decreased from approximately 2.5 x 10(6) Da to approximately 2 x 10(6) Da. Injection of HA did not affect the volume of SF or average molecular weight of HA in samples obtained immediately before each injection or at the end of the experiment, 12 weeks after surgery. The SF HA concentration fell from a baseline value of 2.3 +/- 0.1 mg/ml to 1.1 +/- 0.2 mg/ml the day after surgery and remained low throughout the course of injections. The HA concentration 12 weeks after surgery in the HA injected knees was approximately 40% lower than the preoperative value, although it increased slightly relative to saline injected knees (1.4 +/- 0.3 vs 1.1 +/- 0.01 mg/ml, respectively; p = 0.04). CONCLUSION: Intraarticular injection of HA did not alter the volume of SF or molecular weight of HA in SF of OA canine knees, nor did it restore the HA concentration to that of normal canine SF. PMID- 11409130 TI - Evaluation of glucosamine sulfate compared to ibuprofen for the treatment of temporomandibular joint osteoarthritis: a randomized double blind controlled 3 month clinical trial. AB - OBJECTIVE: To compare the treatment potential of glucosamine sulfate (GS) and ibuprofen in patients diagnosed with temporomandibular joint (TMJ) osteoarthritis (OA). METHODS: Forty women and 5 men received either GS (500 mg tid) or ibuprofen (400 mg tid) for 90 days in a randomized double blind study. ASSESSMENT: TMJ pain with function, pain-free, and voluntary maximum mouth opening, Brief Pain Inventory (BPI) questionnaire and masticatory muscle tenderness were performed after a one week washout and at Day 90. Acetaminophen (500 mg) dispensed for breakthrough pain was counted every 30 days to Day 120. RESULTS: In total, 176 adults were interviewed, 45 (26%) qualified, 39 (87%) completed the study (21 GS, 18 ibuprofen). Four discontinued due to stomach upset (3 ibuprofen, one GS), one due to dizziness (GS), one due to inadequate pain control (ibuprofen). Within group analysis revealed significant improvement compared to baseline of all variables in both treatment groups but no change in acetaminophen used. Fifteen GS (71%) and 11 ibuprofen (61%) improved, with positive clinical response taken as a 20% decrease in primary outcome (TMJ pain with function). The number of patients with positive clinical response was not statistically different between groups (p = 0.73). Between-group comparison revealed that patients taking GS had a significantly greater decrease in TMJ pain with function, effect of pain, and acetaminophen used between Day 90 and 120 compared with patients taking ibuprofen. CONCLUSION: GS and ibuprofen reduce pain levels in patients with TMJ degenerative joint disease. In the subgroup that met the initial efficacy criteria, GS had a significantly greater influence in reducing pain produced during function and effect of pain with daily activities. GS has a carryover effect. PMID- 11409131 TI - Cardiovascular response to upright tilt in fibromyalgia differs from that in chronic fatigue syndrome. AB - OBJECTIVE: To compare the cardiovascular response during postural challenge of patients with fibromyalgia (FM) to those with chronic fatigue syndrome (CFS). METHODS: Age and sex matched patients were studied, 38 with FM, 30 with CFS, and 37 healthy subjects. Blood pressure (BP) and heart rate (HR) were recorded during 10 min of recumbence and 30 min of head-up tilt. Differences between successive BP values and the last recumbent BP, their average, and standard deviation (SD) were calculated. Time curves of BP differences were analyzed by computer and their outline ratios (OR) and fractal dimensions (FD) were measured. HR differences were determined similarly. Based on the latter measurements, each subject's discriminant score (DS) was computed. RESULTS: For patients and controls average DS values were: FM: -3.68 (SD 2.7), CFS: 3.72 (SD 5.02), and healthy controls: -4.62 (SD 2.24). DS values differed significantly between FM and CFS (p < 0.0001). Subgroups of FM patients with and without fatigue had comparable DS values. CONCLUSION: The DS confers numerical expression to the cardiovascular response during postural challenge. DS values in FM were significantly different from DS in CFS, suggesting that homeostatic responses in FM and CFS are dissimilar. This observation challenges the hypothesis that FM and CFS share a common derangement of the stress-response system. PMID- 11409132 TI - Chronic pain and difficulty in relaxing postural muscles in patients with fibromyalgia and chronic whiplash associated disorders. AB - OBJECTIVE: To investigate if muscle tension according to the surface electromyogram (EMG) of the shoulder flexors is increased in consecutive patients with fibromyalgia (FM) or chronic whiplash associated disorders (WAD). METHODS: A total of 59 consecutive patients with FM (n = 36) or chronic WAD (n = 23) performed 100 maximal isokinetic contractions combined with surface electromyography of the trapezius and infraspinatus. A randomized group of pain free female (n = 27) subjects served as control group. Peak torque initially (Pti) and absolute and relative peak torque at endurance level (PTe, PTer) were registered as output variables, together with the EMG level of unnecessary muscle tension, i.e., the signal amplitude ratio (SAR). RESULTS: The patient groups had a higher level of unnecessary tension initially and at the endurance level. The patients had lower absolute output (PTi and PTe), but the relative levels (PTer) did not differ comparing all 3 groups. Subjects with FM had significantly higher body mass index (BMI) than the other groups. BMI did not influence the SAR but correlated positively with PTi. CONCLUSION: The results confirmed earlier findings that groups of patients with chronic pain have increased muscle tension and decreased output during dynamic activity compared to pain-free controls. However, the results indicated there is heterogeneity within groups of patients with the same chronic pain disorder and that not all patients with chronic pain have increased muscle tension. PMID- 11409133 TI - Chronic musculoskeletal pain, prevalence rates, and sociodemographic associations in a Swedish population study. AB - OBJECTIVE: To estimate the prevalence of chronic regional and widespread musculoskeletal pain in a sample of the general adult population and study the association to age, sex, socioeconomic class, immigration, and housing area. METHODS: A cross sectional survey with a postal questionnaire to 3928 inhabitants on the west coast of Sweden. RESULTS: The age and sex adjusted prevalence of chronic regional pain (CRP) was 23.9% and chronic widespread pain (CWP) 11.4% among 2425 subjects who responded to the complete questionnaire. Odds ratio (OR) for CWP showed a systematic increasing gradient with age and was highest in the age group 59-74 yrs (OR 6.36, 95% CI 3.85-10.50) vs age group 20-34 yrs. CWP was also associated with female sex (OR 1.91, 95% CI 1.41-2.61), being an immigrant (OR 1.83, 95% CI 1.22-2.77), living in a socially compromised housing area (OR 3.05, 95% CI 1.48-6.27), and being an assistant nonmanual lower level employee (OR 1.92, 95% CI 1.09-3.38) or manual worker (OR 2.72, 95% CI 1.65-4.49) vs being an intermediate/higher nonmanual employee. OR for CRP showed a systematic increasing gradient with age and was highest in the age group 59-74 yrs (OR 2.22, 95% CI 1.62-3.05) vs age group 20-34 yrs. CRP was also associated with being a manual worker (OR 1.63, 95% CI 1.19-2.23) vs being an intermediate/higher nonmanual employee. CONCLUSION: Chronic musculoskeletal pain is common in the general population. Sociodemographic variables were overall more frequently and strongly associated with CWP than with CRP, which indicates different pathophysiology in the development or preservation of pain in the 2 groups. PMID- 11409134 TI - The influence of work related psychosocial factors and psychological distress on regional musculoskeletal pain: a study of newly employed workers. AB - OBJECTIVE: To determine the influence of short term work related psychosocial factors (work demands, job control, and social support) and psychological distress on regional pain syndromes. METHODS: Newly employed workers were recruited from 12 occupational groups and information collected by questionnaire. Subjects indicated on a blank body manikin any low back, shoulder, wrist/forearm, or knee pain that had occurred during the past month and lasted more than one day. Data were also collected on work related psychosocial factors and on levels of psychological distress [using the General Health Questionnaire (GHQ)]. The relationships between psychosocial factors and psychological distress and each area of pain were calculated as odds ratios with 95% confidence intervals. Adjustment was made for age, sex, and occupational group. RESULTS: 1081 subjects (median age 23; interquartile range 20-27) were recruited to the study shortly after commencing employment: 261 (24%) reported low back pain, 221 (20%) reported shoulder pain, 93 (9%) reported wrist/forearm pain, and 222 (21%) reported knee pain. High levels of psychological distress were associated with increased likelihood of pain, with a trend observed between scores on the GHQ and the odds of pain in each of the 4 sites. Those who perceived their work as stressful most of the time were more likely to report back (OR 1.8, 95% CI 1.01-3.1) or shoulder pain (OR 1.9, 95% CI 1.02-3.4) than those who considered their work seldom stressful. Pace of work or job autonomy was less markedly related to pain at individual sites. Strong relationships were observed between psychological distress, job demands (stressful work, hectic work), low job control, and pain at multiple sites. CONCLUSION: The study has shown that adverse work related psychosocial factors, in particular aspects of job demand and control, influence the reporting of regional musculoskeletal pain. This occurs even after only short term exposure. The odds of reporting these adverse exposures are increased when pain is reported at multiple sites. PMID- 11409135 TI - Unexplainable nondermatomal somatosensory deficits in patients with chronic nonmalignant pain in the context of litigation/compensation: a role for involvement of central factors? AB - OBJECTIVE: To address the prevalence and characteristics of nondermatomal somatosensory deficits (NDSD) in subjects with chronic pain in the context of compensation/litigation. METHODS: Data were collected via standardized history, examination, and patient- as well as physician-drawn body maps in a consecutive series of 194 subjects seen for the purpose of an independent medical examination. RESULTS: Forty-nine subjects (25.3%) with primarily widespread pain (often diagnosed as fibromyalgia) presented with hemisensory or quadrotomal deficits to pinprick and other cutaneous stimuli on the side of lateralized pain or worse pain. The NDSD limbs often had impairment of vibration sense (not infrequently associated with "forehead vibration split"), reduced strength, dexterity or movement, and extreme sensitivity to superficial skin palpation or profound insensitivity to deep pain. Spatial, temporal, qualitative, and evolutionary patterns of NDSD emerged associated with cognitive/affective symptoms. NDSD subjects were more often born outside Canada, more likely to be injured at work, present with abnormal pain behavior, and have negative investigations. CONCLUSION: NDSD are a prevalent problem associated with chronic pain. Future research should explore the prevalence of NDSD in other pain populations, the role of personality and related factors, and the underlying biological substrate of these deficits. PMID- 11409136 TI - Resource utilization and cost of rheumatic fever. AB - OBJECTIVE: The socioeconomic effects of rheumatic fever (RF) in Brazil, including direct and indirect costs to patients and their families and to society, are largely unknown. We evaluated the utilization of resources and costs related to RF in a tertiary center caring for low income patients in the city of Sao Paulo, Brazil. METHODS: One hundred patients with RF, younger than 18 yrs, with followup of at least one year, were sequentially selected to provide complete information on a questionnaire. Additional data were collected from patients' charts. The utilization of resources was evaluated for each patient throughout the entire disease course. Costs were determined for patients and their families as well as for the society, using variables from 3 different systems: the national public health system, used by most lower income groups; the Brazilian Medical Association, which regulates charges and fees utilized by health plans and insurance companies; and costs charged by private practitioners, paid directly by patients. RESULTS: The RF population studied belonged to a low socioeconomic level. The mean monthly family income was $625.20 US. The mean disease duration was 3.9 yrs (range 1-10). Patients had a total of 1657 medical consultations, 22 hospital admissions, and 4 admissions to intensive care unit. Work absenteeism among parents was calculated as 22.9%, equivalent to 901 days of missed work; about 5% of the parents lost their jobs. Patients showed a high rate of school failure (22%). Considering the public system as a reference, direct, indirect, and total costs to society per 100 patients throughout the entire disease duration were $105,860 US ($271/patient/yr), $18,803 US ($48/patient/yr), and $124,663 US (US $319/patient/yr), respectively. When health care plan and private systems were taken as reference, the total costs were $423,550 US and $684,351 US, respectively. CONCLUSION: RF and rheumatic heart disease have an important socioeconomic impact in Brazil; costs of RF made up roughly 1.3% of annual family income. The estimated annual cost of RF for society in Brazil is $51,144,347.00 US. PMID- 11409137 TI - The hip or not. AB - Pathologies within the ileopsoas compartment can present as hip pain and as such can be mistaken for diseases of the hip joint. Radiological investigations are useful in helping to differentiate between these pathologies. PMID- 11409138 TI - Periarticular calcinosis associated with anti-Jo-1 antibodies sine myositis. Expanding the clinical spectrum of the antisynthetase syndrome. AB - We describe a 58-year-old woman who developed interstitial lung disease (ILD), polyarthritis, and anti-Jo-1 antibodies, with no clinical evidence of myositis. Despite successful treatment with corticosteroid and azathioprine for her arthritis and pulmonary condition, she developed deforming arthropathy of the hands, with periarticular calcinosis. The association of anti-Jo-1 antibodies, ILD, and periarticular calcinosis with subluxing arthropathy sine myositis is rare, with few cases reported. This report expands the clinical spectrum of the antisynthetase syndrome, which is broader than previously reported. PMID- 11409139 TI - Malignant hyperthermia susceptibility revealed by myalgia and rhabdomyolysis during fluoroquinolone treatment. AB - Fluoroquinolones cause myalgia, but this complication is not clearly documented. We describe a patient who developed myalgia and rhabdomyolysis during fluoroquinolone treatment. The patient was a 33-year-old man treated with norfloxacin for common cystitis. He complained of general muscular fatigue, tendon disorders, and articular pain during treatment. When the antimicrobial agent was stopped, symptoms decreased, with persistence of slight myalgia for 10 days. Rhabdomyolysis was detected. Six months later, investigation by 31P magnetic resonance spectroscopy revealed an oxidative disorder and an abnormal abundance of phosphomonoesters. In vitro contracture tests led to a diagnosis of malignant hyperthermia susceptibility. Our case shows that for any subject presenting myalgia with rhabdomyolysis triggered by fluoroquinolone treatment, the presence of a latent myopathy should be investigated. PMID- 11409140 TI - Lower limb giant cell arteritis and temporal arteritis: followup of 8 cases. AB - Among 8 patients with giant cell arteritis (GCA) (6 women, 2 men) whose clinical presentations were compatible with temporal arteritis (TA), 6 were followed for 37-105 (mean 74.9) months, one died shortly after treatment onset, and the last was asymptomatic (10 mg steroids/day) when lost to followup at 29 months. All 8 patients had bilateral leg claudication of recent onset; for 6 patients, this was the first symptom. All leg angiograms showed multiple, bilateral, long and smooth stenoses, thromboses, or both. Biopsies of diseased leg arteries from 4 patients provided histological proof of GCA; another case was histologically proven post mortem. Among the 5 patients who met at least 3 American College of Rheumatology criteria of GCA or TA, 3 without histologically documented leg GCA also had biopsy proven temporal GCA (n = 1), or headaches and claudication and angiographic inflammatory arteritis of the arms (n = 2). All patients received steroids; 3 had bypasses, one with endarterectomy. Five are asymptomatic after 24 100 months of steroids (mean 50.6). Revascularization was not successful; one amputation was necessary. Large artery involvement in GCA can affect the legs. Bilateral and rapidly progressive intermittent claudication of recent onset is the most common symptom, even in the absence of headaches or the presence of a silent inflammatory syndrome. Early diagnosis allows rapid initiation of steroid therapy, which is usually able to generate a sufficiently good response to avoid vascular surgery. PMID- 11409141 TI - Consensus recommendations for the assessment and treatment of rheumatoid arthritis. PMID- 11409142 TI - Management of rheumatoid arthritis: the historical context. AB - We review the historical highlights of the management of rheumatoid arthritis (RA). Studies of nonsteroidal antiinflammatory drugs, disease modifying antirheumatic drugs, and biological agents over 5 decades were evaluated and summarized. There is emphasis on drug therapy as it has developed and evolved from empirical relief of symptoms with salicylates to targeted intervention in the immunoinflammatory process with tumor necrosis factor inhibitors. A therapeutic paradigm has been proposed to rationalize the use of the available therapies. If one accepts the thesis that both the acute and chronic consequences of RA are due to persistent misdirected and inadequately controlled inflammation that causes tissue destruction and loss of function, then prolonged complete control of the abnormal inflammatory process is the fundamental first step in the management of all patients with RA. Unfortunately, even with the newest therapeutic options to treat RA, most patients achieve only partial suppression of inflammation and many lose therapeutic benefit after an initial good response. The management of persistent or recurrent rheumatoid inflammation and disability continues to be a challenge. It remains to be determined whether the future addition of more potent specific interventions in the immunoinflammatory process will be able to solve this problem without disarming host defenses against infections and tumors. PMID- 11409143 TI - Evaluating severity and status in rheumatoid arthritis. AB - There is general agreement regarding the most appropriate examinations and methods to use to evaluate change in status in randomized controlled trials (RCT). However, no guidelines exist to aid in determining and evaluating actual status rather than change in status, particularly when applied to individual patients with rheumatoid arthritis (RA). In addition, methods appropriate for clinical trials may not be useful in evaluating individual patients because of time constraints. This report reviews current methods of evaluation and develops modified methods, based on data bank research that will be useful in clinical practice and in the evaluation of RCT and observational studies. Using data from longitudinal observational data banks, further reduction in the number of joints examined is evaluated to reconcile the time constraints of clinical practice with the need to maintain reliability and validity. Percentile methods to determine severity status are applied to the variables used in RCT and extended further to observational studies and routine clinical practice. Shortened joint counts, based on modifications of the Ritchie method, are identified that allow for examination of groups of 18 (clinical-18) and 16 (clinical-16) joints, the clinical-16 omitting the metatarsophalangeal joints. Using percentile charts, actual severity valuations are given to the variables evaluated in the clinic as well as in RCT. Disease activity status of clinic patients can be determined quantitatively thus allowing clinicians further insight into the status and prognosis of their patients. By quantifying disease activity severity, clinicians and 3rd party payers can better evaluate the appropriateness of and response to disease modifying antirheumatic drugs and biologic therapies. Further, RCT can be evaluated as to severity status of patients participating, and the generalizability of RCT can be better evaluated. PMID- 11409144 TI - Patterns of drug use in rheumatoid arthritis. PMID- 11409146 TI - New thoughts on old bones. PMID- 11409145 TI - Patterns of drug use in rheumatoid arthritis. PMID- 11409147 TI - HLA-B27 associated spondyloarthropathy and severe ascending aortitis. PMID- 11409148 TI - Development of systemic lupus erythematosus following autologous bone marrow transplant for acute lymphocytic leukemia. PMID- 11409149 TI - Morning stiffness: how common is it and does it correlate with physician and patient global assessment of disease activity? PMID- 11409150 TI - Femoral head necrosis and hyperhomocysteinemia. PMID- 11409151 TI - Overview of current therapies for rheumatoid arthritis. PMID- 11409152 TI - Disease modifying antirheumatic drugs: longterm safety issues. AB - The trend for more aggressive management of rheumatoid arthritis includes earlier introduction of disease modifying antirheumatic drugs (DMARD). As patients may continue their therapy for several decades instead of several years, the evaluation of benefit versus risk of DMARD with particular emphasis on longterm safety is essential. Longterm safety assessment is difficult for a number of reasons: there are relatively few trials that have followed patients beyond 5 years and the use of a combination of DMARD therapy with nonsteroidal antiinflammatory drugs and corticosteroids complicates the assessment of an observed adverse event with a particular drug. This review of longterm studies incorporating DMARD provides insight into adverse events associated with currently available DMARD. PMID- 11409153 TI - Disease controlling antirheumatic therapy in spondyloarthropathy. AB - The management of spondyloarthropathy (SpA) is based more on clinical presentation (axial vs peripheral involvement) than on actual disease diagnosis. Treatment is aimed at controlling inflammation, ankylosis, and abnormal posture. Nonsteroidal antiinflammatory agents are the cornerstone of treatment for patients with axial involvement. If these do not work, disease controlling antirheumatic therapy is usually prescribed. Several drugs, particularly sulfasalazine, produce clinically significant improvement in patients with articular peripheral involvement. In contrast, the therapeutic options for patients with refractory axial involvement are very limited. For this group, "potential" (pamidronate, thalidomide) or "specific" (infliximab) tumor necrosis factor blockers are of potential interest and deserve further evaluation. Therapeutic efficacy is monitored chiefly according to clinical variables, although biochemical markers such as C reactive protein are also helpful. PMID- 11409154 TI - Conventional DMARD options for patients with a suboptimal response to methotrexate. AB - Methotrexate (MTX) is one of the disease modifying antirheumatic drugs (DMARD) commonly used to treat rheumatoid arthritis (RA). However, MTX therapy alone rarely results in remission and frequently does not even produce 50% improvement. Therefore, over the course of their disease, many patients will require additional therapy to manage their clinical symptoms. A number of treatment options have proven effective for such patients, most of which entail the continuation of MTX therapy and the addition of other DMARD. Although the combination of MTX and hydroxychloroquine (HCQ) is the one most commonly used in the US, many clinicians (particularly in Europe) prefer the combination of MTX and sulfasalazine. In addition, excellent data now exist for the triple combination of MTX, HCQ, and sulfasalazine in patients who have had a suboptimal response to MTX, as well as in those with early or well established disease. Other combinations, including MTX + cyclosporine or leflunomide, have also been helpful in some patients. Most recently, the tumor necrosis factor blockers, etanercept and infliximab, have successfully been used to treat a number of patients resistant to MTX. The combination of MTX with DMARD or biological agents with different mechanisms of action greatly expands the treatment options for patients with RA. PMID- 11409155 TI - How do the biologics fit into the current DMARD armamentarium? AB - Most disease modifying antirheumatic drugs (DMARD) are discontinued within 5 years because of loss of clinical efficacy or toxicity. As a result, there has been a concerted effort to develop new immunomodulatory agents, particularly biological agents, that block the putative proinflammatory cytokines. Among the agents developed thus far, inhibitors of tumor necrosis factor (TNF) have shown perhaps the greatest promise as therapeutic agents for rheumatoid arthritis (RA). Two TNF-blocking agents, etanercept (Enbrel) and infliximab (Remicade), have been approved in the US and more recently in Europe, for the treatment of patients with RA. The results of randomized placebo controlled trials have shown that both agents significantly decrease the intensity of synovitis and prevent or retard the progression of cartilage destruction, especially when combined with methotrexate. Their side effect profiles appear to be acceptable, although rare cases of lupus-like diseases and of severe infections have been reported. Although the early clinical experience with these agents has been encouraging, their longterm safety and continuing efficacy in the general population with RA, as well as in high risk patient subsets (i.e., patients with malignancies or chronic infections), remain to be determined. In addition, the costs of these newer agents must be justified on clinical grounds. Because of the questions still surrounding these new treatment principles, several consensus conferences have been held in Europe and the US to address the role of the new biologicals in the current RA armamentarium. PMID- 11409156 TI - Treating early rheumatoid arthritis in the younger patient. AB - Early diagnosis and intervention may provide the greatest hope for reducing the disability associated with rheumatoid arthritis (RA). In patients with early RA, accurate diagnosis can be delayed by limited access to a specialist service, slow evolution of the clinical features, and lack of definitive diagnostic criteria. However, acute phase reactants, serologic features including presence of rheumatoid factor, and immunohistologic analysis of synovial tissue can provide the basis for differentiating RA from other forms of arthritis. Factors associated with poorer prognosis in patients with early RA are female sex, larger number of joints involved, elevated levels of acute phase reactants, presence of rheumatoid factor, and radiologic evidence of joint damage. Special treatment considerations in younger persons with RA include issues related to conception, pregnancy, and lactation. Methotrexate, hydroxychloroquine, sulfasalazine, and low dose corticosteroids are usually the mainstays of treatment for younger patients with RA. Recommendations for taking these drugs while considering conception vary with their effect on fertility and on the developing embryo. Sulfasalazine, for example, can be taken during pregnancy but caution is advised for breastfeeding mothers. Leflunomide must be discontinued for 2 years before attempting conception; this time can be shortened if the patient opts for drug washout. PMID- 11409157 TI - Polyunsaturated fatty acids influence neonatal monocyte survival. AB - The n-3 and n-6 polyunsaturated fatty acids (PUFAs) are essential dietary constituents. They are potent modulators of the human immune response, and research has endeavoured to optimise the ratio of n-3 to n-6 fatty acids in the lipid emulsion component of total parenteral nutrition to harness their beneficial effects in the clinical setting. PUFAs modulate apoptosis of certain tumour cells and cell lines. Monocytes, which are major effector cells of the innate immune system, play a central role in the initiation, development, and outcome of the immune response. They are crucial in the defence against invading pathogens and are involved in the lysis of infected or malignant cells, wound healing, repair, and remodeling of tissues. In the present study we investigated whether PUFAs might evoke apoptosis in newborn monocytes. Purified cord-blood monocytes collected from uncomplicated full-term pregnancies were incubated for 24 h in complete medium in the presence or absence of one of the n-3 PUFAs docosahexaenoic acid (DHA) and eicosapentaenoic (EPA) or the n-6 PUFA arachidonic acid (AA). Following incubation, cells were triple-labelled with annexin V, CD14, and propidium iodide prior to flow-cytometric analysis to determine the degree of cell death. All experiments were performed in triplicate and data expressed as mean +/- 1 S.D. (%). In the absence of fatty acids, 30 +/- 4% of control cord monocytes underwent apoptosis or necrosis after 24 h incubation. At a concentration of 50 microM, none of the PUFAs had a significant effect on monocyte cell death, but at a dose of 100 microM, DHA resulted in 60 +/- 4% cell death (P < 0.05) while the other PUFAs had no significant effect. In contrast, at higher concentrations (200 microM), all the PUFAs significantly increased monocyte cell death (AA: 70 +/- 5%, DHA: 86 +/- 2%, EPA: 70 +/- 4%). PUFAs thus exert a potent influence on cord monocyte cell survival in vitro. Their effect is dose-dependent and DHA appears to be the most potent of the fatty acids tested. The influence of PUFAs on neonatal monocyte-cell survival suggests a novel mechanism whereby PUFAs may modulate the immune response. PMID- 11409158 TI - Effect of low fat diet on lipid absorption and fatty-acid transport following bowel resection. AB - Low-fat diets (LFD) are used extensively in many different clinical conditions. However, the effect of this diet on lipid absorption and cellular long-chain fatty-acid (LCFA) transport is unknown. Fatty-acid translocase (FAT), the rat homologue of human CD36, is one of several LCFA plasma-membrane transport proteins that may play an important role in intestinal lipid uptake. The purpose of this study was to investigate the effects of a LFD on intestinal expression of FAT/CD36, enterocyte fatty-acid transport, and in-vivo lipid absorption in rats following bowel resection. Adult male Sprague-Dawley rats were divided into five experimental groups: normal rats fed normal chow(NR-NC) (10 kcal% fat), normal rats fed a LFD (NR-LFD) (3 kcal% fat), sham rats fed normal chow (Sham-NC), short bowel syndrome rats fed normal chow (SBS-NC), and SBS rats fed a LFD (SBS-LFD). SBS rats underwent 75% small-bowel resection, while sham animals underwent bowel transection and reanastomosis. Food intake, fecal mass, and fecal fat were measured over the last 3 days before death on day 14. Final body weight, plasma lipids and protein, and tissue total lipids in liver, adipose tissue, and intestine were determined at death. Total RNA from the mucosa of the duodenum, jejunum, and ileum was extracted for Northern blot analysis to determine fatty acid translocase (FAT)/CD36 mRNA levels. An established cellular LCFA transport assay was used to determine isolated enterocyte [3H]-oleate uptake. Students t test was used to determine statistical significance (P < 0.05). NR-LFD rats demonstrated a small increase in overall food absorption and no change in fat absorption compared to NR-NC animals. A significant decrease in FAT/CD36 mRNA levels was seen in the duodenum and jejunum in NF-LFD rats (vs NR-NC) and was accompanied by reduced LCFA transport by isolated enterocytes from the jejunum and ileum. SBS-LFD rats demonstrated decreased FAT/CD36 mRNA levels in all three segments and a concomitant decrease in LCFA uptake enterocytes compared to the SBS-NC group. In addition, SBS-LFD rats showed significantly lower final body weight and plasma lipids compared to SBS-NC animals. PMID- 11409159 TI - Effect of probiotics on enterocyte bacterial translocation in vitro. AB - Enteral probiotics such as Lactobacillus casei GG (LGG) have been used in the treatment of a variety of intestinal disorders in infants and children, including diarrhea, malabsorption, and Clostridium difficile colitis. We have previously demonstrated that the probiotic bacterium LGG has an inhibitory effect on bacterial translocation (BT) in a neonatal rabbit model. However, this in-vivo model is limited for investigating the cellular and molecular mechanisms responsible for probiotic inhibition of BT. The purpose of this study was to determine the efficacy of LGG in reducing the rate of Escherichia coli C25 (E. coli C25) translocation using an in-vitro enterocyte cell-culture model. Human colonic carcinoma (Caco-2) enterocytes were seeded in porous filters in the apical chamber of a two-chamber cell-culture system and grown for 14 days to confluence. The monolayers were incubated at 37 degrees C with LGG for 180 min. Non-adherent LGG was washed away prior to a 120-min incubation period with 10(5) CFU E. coli C25. E. coli that had translocated across the enterocyte monolayer were quantified by growing basal-chamber media samples on gram-negative bacteria specific MacConkey's agar. In order to determine monolayer integrity, transepithelial electrical resistance (TEER) was measured across Caco-2 cells treated with LGG and E. coli. Statistical analysis was by ANOVA with P < 0.05 considered significant. LGG inhibited E. coli translocation at all LGG concentrations tested. The TEER ratio was not significantly altered by addition of LGG or E. coli (0.9 +/- 0.03 vs 0.8 +/- 0.05). These results demonstrate that the probiotic bacterium LGG inhibits BT of E. coli C25 in a dose-dependent manner in an in-vitro cell-culture model. This model should be valuable in investigating the cellular and molecular mechanisms involved in the inhibition of pathological enteral bacteria by probiotic agents. PMID- 11409160 TI - The effect of phospholipids and fatty acids on tight-junction permeability and bacterial translocation. AB - The activity of phospholipase A2 (PLA2) is elevated in the intestinal epithelia of patients with inflammatory bowel disease (IBD). We recently reported that PLA2 mediates hydrolysis of phosphatidylcholine (PC) to lysophosphatidylcholine (L-PC) when both are applied to the apical surface of cultured EC monolayers, resulting in increased bacterial translocation (BT) and decreased transepithelial electrical resistance (TEER). Free fatty acids (FFA) are the other products of this reaction, however, their effect on Caco-2 cell permeability has not been reported. In addition to PC, other luminal phospholipids are present at the surface of the enterocyte. PLA2 may also mediate the hydrolysis of luminal phospholipids other than PC. The aim of this study was to examine the effects of phospholipids other than PC and common FFA on intestinal epithelial permeability and BT. Human Caco-2 enterocytes were grown to confluence on porous filters in the apical chamber of a two-chamber cell-culture system. Monolayer integrity and tight-junction permeability were measured as TEER. First, common FFA released by PC hydrolysis were determined using thin-layer chromatography (TLC). In separate experiments, monolayers were treated with phosphatidylethanolamine (PE), lysophosphatidylethanolamine (L-PE), or palmitoleic acid, oleic acids, linoleic acids, and arachidonic acid solubilized in solution with PC. The magnitude of BT was determined 2 h after treatment by adding Escherichia coli C25 to the apical chamber followed by quantitatively culturing basal-chamber samples. Statistical analysis was by the Kurosaki-Wallis test. TLC of PC samples incubated with PLA2 on the apical surface of Caco-2 monolayers demonstrated the production of palmitoleic acid, oleic acids, linoleic acids, and arachidonic acid. L-PE significantly decreased TEER compared to controls, but to a lesser degree than L PC alone. L-PE had no effects on BT. Palmitoleic acid and oleic acid likewise significantly decreased TEER compared to controls, however, less than L-PC. All FFA tested had no effect on BT. Phospholipids applied to the apical surface of enterocytes, such as those found in vivo in mucus, can be hydrolyzed by the enzyme PLA2 resulting in lysophospholipid and FFA species that can alter enterocyte monolayer permeability. However, FFA and L-PL, other than L-PC, appear to have no effect to stimulate BT. This observation may have clinical implications in the pathogenesis and treatment strategies for IBD patients in whom enterocyte PLA2 activity has been shown to be elevated. PMID- 11409161 TI - Effect of secretory immunoglobulin A on bacterial translocation in an enterocyte lymphocyte co-culture model. AB - Intestinal secretory immunoglobulin A (sIgA) plays an important role in gut mucosal immunity in vivo; however, in-vitro enterocyte models for studying the mechanisms of these effects are lacking. This study utilizes a cell-culture model to investigate the effect of sIgA on bacterial translocation (BT) across human enterocytes co-cultured with human lymphoid cells (Raji cells). This model is intended to mimic in-vivo enterocyte/lymphocyte interactions found in intestinal follicle-associated epithelia. Human Caco-2 enterocytes were grown to confluence on porous filters in the apical chamber of a two-chamber cell-culture system. After differentiation, human B lymphoid cells (Raji cells) were added to the basolateral surface of Caco-2 monolayers for 3 days' co-culture, followed by washing away of unincorporated Raji cells. Transepithelial electrical resistance (TEER) was used to measure tight-junction permeability. Monolayers were treated with or without sIgA, IgG (negative control), or mannose (positive control). BT across the cell monolayer was determined 1.5 h after addition of Escherichia coli. Statistical analysis was by the Kruskal-Wallis test, P below 0.05 considered significant. In co-culture monolayers treated with sIgA, IgG, or mannose, there was no significant effect on TEER; however, the magnitude of BT across cells treated with sIgA (1.3 +/- 0.4 log10CFU/ml) and mannose (1.6 +/- 1.1 log10CFU/ml) was significantly decreased compared to PBS (3.9 +/- 0.4 log10CFU/ml) and IgG (2.9 +/- 0.6 log10CFU/ml) controls (P < 0.05). sIgA BT inhibition was dose-dependent. BT inhibition by sIgA and mannose was additive (0.5 +/- 1 log10CFU/ml). Inhibition of BT was negated when sIgA and mannose were removed by washing prior to E. Coli addition (3.6 +/- 0.5 log10CFU/ml), suggesting that both inhibitors act through bacterial binding. PMID- 11409162 TI - Tissue antioxidant capacity and bacterial translocation under total parenteral nutrition. AB - Alterations in the antioxidative system have been observed during total parenteral nutrition (TPN). Light exposure or changes in the composition of TPN formulas may affect this system. Bacterial translocation (BT) is frequent under TPN and may be related to oxidative status. The aim of this study was to determine the adverse effects of standard and glutamine-enriched TPN, with or without light exposure, on oxidative status (liver and kidney-reduced glutathione, GSH) and its relationship to BT. Thirty-three adult Wistar rats underwent central-venous cannulation and were randomly assigned to one of four groups receiving different TPN regimes for 10 days. The TPN group (n = 10) had standard TPN, the TPN(-) group (n = 8) standard TPN without light exposure, the GTPN group (n = 8) glutamine-enriched TPN, and the GTPN(-) group (n = 7) glutamine-enriched TPN without light exposure. A sham group (n = 16) receiving chow and water ad libitum and saline i.v. served as controls. At the end of the experiment, GSH was determined in liver and kidney tissue. Mesenteric lymph nodes and peripheral and portal blood samples were cultured for BT. Compared to sham rats, TPN groups had statistically significant lower GSH levels, but there were no differences between standard or glutamine-enriched groups or light-exposure groups. Sham animals had 12% BT. Significantly higher BT (P < 0.05) occurred in TPN rats: 70% in the TPN group, 88% in the TPN(-) group, 86% in GTPN (-) animals, and only 50% in the GTPN group (P = 0.06 vs TPN group). In conclusion, (1) TPN reduces antioxidant capacity; (2) glutamine supplementation or light protection does not improve tissue antioxidant capacity under TPN; (3) the absence of light exposure does not improve TPN-related BT; and (4) glutamine supplementation tends to reduce BT only in the presence of light. PMID- 11409163 TI - Gene expression of insulin-like growth factor-1 and epidermal growth factor is downregulated in the heart of rats with nitrofen-induced diaphragmatic hernia. AB - Newborns with congenital diaphragmatic hernia (CDH) still have high mortality. Recently, a possible role of cardiac maldevelopment has been suggested. Human and experimental studies have demonstrated that heart weight is significantly reduced in the presence of CDH. Recent studies have suggested an important role for insulin-like growth factor-I (IGF-I) in the regulation of cardiac growth, structure, and function. Administration of IGF-I to normal rats has been shown to cause cardiac hypertrophy. Epidermal growth factor (EGF) plays an important role in cardiac differentiation and development. The aim of this study was to determine the gene-level expression of IGF-I and EGF in the hearts of rats with nitrofen-induced CDH using the reverse-transcription polymerase chain reaction technique (RT-PCR). CDH was induced in pregnant rats following administration of 100 mg nitrofen on day 9.5 of gestation (term 22 days). In control animals, the same dose of olive oil was given without nitrofen. Cesarean section was performed on day 21 of gestation. The fetuses were divided into three groups: normal controls (n = 8), nitrofen without CDH (n = 8), and nitrofen-induced CDH (n = 8). Total RNA was extracted from the hearts in each group and measured. mRNA was extracted from total RNA. RT-PCR was performed to evaluate mRNA expressions of IGF-I and EGF. Levels of mRNA were expressed as a ratio of band density divided by that of beta-actin, a housekeeping gene known to be expressed at a constant level. IGF-I mRNA expression was significantly decreased in CDH hearts (0.177 +/- 0.109) compared to controls (0.393 +/- 0.138) (P < 0.01) and nitrofen hearts without CDH (0.321 +/- 0.088) (P < 0.05). EGF mRNA expression was significantly decreased in CDH hearts (0.218 +/- 0.118) compared to controls (0.534 +/- 0.196) (P < 0.01) and nitrofen hearts without CDH (0.383 +/- 0.136) (P < 0.05). Decreased cardiac gene expression of IGF-I and EGF in the hypoplastic heart suggests that cardiac hypoplasia in nitrofen-induced rat CDH may be due to reduced synthesis of IGF-I and EGF by myocytes in the developing heart. PMID- 11409164 TI - Does the developing liver inhibit early lung growth in congenital diaphragmatic hernia? AB - It has been hypothesised that the liver induces lung hypoplasia in congenital diaphragmatic hernia (CDH) by non-compressive intrathoracic growth rather than traditional mass herniation. Utilising a co-culture system, we tested the capacity of liver cells to inhibit lung growth by contact rather than compression. Heart, liver, and lungs were microdissected from normal rat embryos (n > 20 from at least three litters) on day 13.5 of gestation. Monolayer cultures of enzymatically dispersed livers and hearts were established at the same cell density. Lung primordia were cultured in direct contact with hepatic cells or partitioned from them by a permeable polytetrafluoroethylene membrane. This permits the contributions of diffusable factors and cell contact to be distinguished. Lungs were similarly cultured in direct contact with or partitioned from cardiac cells. Lungs cultured in isolation served as further controls. Daily inspection permitted assessment of in-vitro lung growth. Growth of lungs in direct contact with hepatic cells was equivalent to that of lungs partitioned from liver cells. Lungs in direct contact with cardiac cells and lungs partitioned from cardiac cells were also not inhibited compared to lungs cultured in isolation. Early lung development is thus not inhibited by humoral or contact-mediated interactions with embryonic liver cells. Lung hypoplasia in CDH is therefore unlikely to originate from contact inhibition with the developing liver. An intrinsic pulmonary defect may better explain hypoplastic lung development in CDH. PMID- 11409165 TI - Neural crest-derived defects in experimental congenital diaphragmatic hernia. AB - Congenital diaphragmatic hernia (CDH) is often associated with other malformations. This study tests the hypothesis that the heart and great vessels, thymus, parathyroids, and thyroid might be abnormal in the rat model of CDH as a result of disturbed neural-crest development. Time-mated pregnant rats were fed either 100 mg 2-4-dichlorophenyl-p-nitrophenyl ether (nitrofen) or vehicle on gestational day 9.5. Diaphragm, lung, heart, and thymic malformations were sought after dissection and the parathyroids and thyroid were histologically investigated in term fetuses. Ten control fetuses had no malformations, whereas 22 of 32 nitrofen fetuses had CDH and 20 had cardiovascular defects like narrow pulmonary outflow tract (n = 7), aberrant right subclavian artery (n = 7), ventricular septal defect (n = 4), atrial septal defect (n = 4), tetralogy of Fallot (n = 2), double-outflow right ventricle (n = 2), right ductus arteriosus (n = 2), and others. The thymus was present but was significantly hypoplastic in all nitrofen fetuses and was ectopic or single-lobed in 28% of them while the parathyroid glands were unilaterally absent or ectopic in 50%. The thyroid was only minimally malformed or ectopic. In conclusion, malformations of structures derived from the pharyngeal arches are likely neural-crest related in rats exposed to nitrofen. PMID- 11409166 TI - Cell-adhesion molecules and fibroblast growth factor signalling in Hirschsprung's disease. AB - Hirschsprung's disease (HD) is characterised by the absence of ganglion cells and the presence of hypertrophic nerve trunks in the distal bowel. It has been suggested that aganglionosis may be caused by failure of differentiation as a result of microenvironmental change after neuronal migration has occurred. Recently, it was reported that cell-adhesion molecules (CAMs) and fibroblast growth factors (FGFs) stimulate neurite outgrowth through activation of FGF receptors (FGFRs) in neurons. The aim of this study was to investigate the expression of CAMs FGFs, and FGFRs in ganglionic (NG) and aganglionic (AG) segments of HD in order to understand the role of CAM-FGF signalling in the pathogenesis of HD. Specimens from NG and AG segments of bowel from 11 patients with HD were obtained at the time of definitive pull-through operation, snap frozen in OCT compound, and stored at -70 degrees C. Aganglionosis was confirmed by Haematoxylin and eosin staining and acetylcholinesterase histochemistry; 8 micron cryosections were immunostained using the standard streptavidinbiotin immunoperoxidase method. The following antibodies were used as the first antibody; FGF2 and FGF7 for FGFs, FGFR1 and FGFR2 for FGFRs, NCAM, L1CAM, and N cadherin for CAMs. FGF2, FGF7, and FGFR2 were expressed in neuronal tissue of NG segments as well as in hypertrophic nerves of AG segments. There was a lack of FGFRI expression in neuronal tissue of both NG and AG bowel. Immunoreactivity with all three CAMs was detected in ganglion cells in NG bowel and in hypertrophic nerve trunks in AG bowel. In contrast the numbers of CAM-positive nerve fibres in muscle layers were markedly decreased in AG bowel compared to NG bowel. The markedly decreased expression of CAMs on nerve fibres within the muscle of AG bowel suggests that CAM-FGF signalling is altered in HD, resulting in failure of enteric neuroblast migration. PMID- 11409167 TI - Assessment of protein gene product 9.5 as a marker of neural crest-derived precursor cells in the developing enteric nervous system. AB - The neurons and glial cells of the enteric nervous system (ENS) are derived from the neural crest. To study the developmental events involved in congenital abnormalities of the ENS, it is essential to identify all neural-crest cells (NCC) in the prenatal gut. The low-affinity neurotrophin receptor p75 is currently considered to be a gold-standard marker, but because it is a membrane protein, it is lost during procedures that permeabilise cells that are necessary to identify intracellular components and in apoptosis and cell-proliferation assays. We have therefore assessed the potential of the intracellular neuronal marker protein gene product (PGP) 9.5 as a label for neural-crest-derived precursor cells during gut development. Gut was taken from mouse embryos at 11.5 days post-coitum, at which time NCC had reached the proximal colon. Cellular p75 and PGP9.5 expression was determined by double-labelling immunofluorescence. The leading edge of neural-crest migration was defined as the 10 most distal p75 labelled cells. The neuronal marker PGP9.5 labelled NCC as they migrated along the gut at day 11.5. At the leading edge of migration, over 95% of p75-positive cells also expressed PGP9.5, and all PGP9.5-positive cells were also labelled for p75. PGP9.5 is expressed by at least 95% of neural-crest-derived precursor cells at the leading edge of migration along the gut. Thus, it can be used as a robust marker for developing NCC in the gut. PMID- 11409168 TI - Angiotensin-converting enzyme genotype distribution in familial vesicoureteral reflux. AB - Vesicoureteric reflux (VUR) is known to occur in families. In siblings of index patients with VUR, there is a much higher incidence (16% to 46%) than in the general population. The renin-angiotensin system plays an important role in renal development. Recently, it has been reported that angiotensin-converting enzyme (ACE) I/D gene polymorphism is a risk factor for renal parenchymal damage in patients with congenital urological abnormalities. The aim of this study was to investigate the ACE I/D genotype pattern in familial VUR patients. Blood samples were obtained from 86 families in which two or more members had VUR. Samples of DNA were extracted from 407 blood samples (183 affected patients and 224 non affected family members). To detect ACE I/D polymorphism, polymerase chain reaction (PCR) amplification was performed using specific primers for the ACE gene. PCR products were electrophoresed with 2% agarose gel containing ethidium bromide. Among 224 non-affected family members the ACE genotype distribution of DD, ID, and II was 23%; 56% and 21%, respectively. The ACE genotype distribution of 183 affected patients was 28%, 47% and 25%, respectively. There was no significant difference in ACE I/D distribution between affected patients and their non-affected families. Both the ACE genotype distribution of affected patients and that of non-affected family members were not significantly different from the previously reported genotype distribution of the normal Caucasian population. This study demonstrates that ACE genotype frequencies are similar in index patients with VUR and their unaffected siblings, thus suggesting that the ACE gene is not involved in the development of familial VUR. PMID- 11409169 TI - Enlargement of the processus vaginalis during testicular descent in rats. AB - The role of the processus vaginalis (PV) during inguinoscrotal testicular descent remains controversial. Some authors propose passive dragging of the PV by the migrating testis, while others suggest active elongation. In addition, the exact site of growth is unknown. Our aim was to determine whether the PV actively proliferates at its tip or stretches passively during the inguinoscrotal phase of descent in the rat. Gubernacula were removed from Sprague-Dawley (SD) rats and congenitally-cryptorchid TS mutants. Animals (at days 3, 7, 10, and 11) were treated with bromo-uridine deoxyribose (BUdR) 2 h before death. BUdR incorporation into newly-synthesised DNA served as a marker for cell division. The gubernacula were processed for haematoxylin and eosin and immunoperoxidase staining. Three sites were examined: (1) the tip of the PV on either side of the gubernacular bulb; (2) the proximal gubernacular cord; and (3) the proximal parietal PV. At each site, 50 adjacent cells were counted and the number of positive cells recorded. The highest BUdR labelling in SD rats was at the tip (site 1) on day 3 (17/50) compared with sites 2 (11/50) and 3 (9/50) (P < 0.05). Labelling decreased by 7 and 11 days to similar levels in all three sites. In TS rats, labelling rates were lower at day 3 and were highest at the tip at day 11. These results suggest active growth of the caudal tip of the PV during testicular descent. In normal rats, the growth rate slows as the testis approaches the scrotum. By contrast, in TS rats growth continued longer. We propose that the PV elongates actively from the tip to allow the intraperitoneal testis to leave the abdomen in a special peritoneal diverticulum. PMID- 11409170 TI - Evaluation of developing rat testis by phosporus 31P magnetic resonance spectroscopy and DNA flowcytometry. AB - For the assessment of germ-cell maturation of the seminiferous tubules, DNA flowcytometry is a rapid and sensitive method. Phosporus 31P magnetic resonance spectroscopy (MRS) is a non-invasive alternative that demonstrates the metabolic status of the testis, reflecting the type and relative proportion of germ cells in the testis. This study was designed to evaluate the utility of 31P MRS in reflecting the haploid-cell population of the testis as measured by DNA flowcytometry. A single testicle was evaluated in male Wistar pre-pubertal rats at 30, 40, 50, and 60 days of age. In order to minimize the contamination of signals from the contralateral testis, scrotum, and tail, the technique was modified and the testis was evaluated ex-vivo with an intact blood supply. At 30 days of age the percentage of haploid cells was 43.6 +/- 1.8, and this increased to 72.7 +/- 1.4 at 60 days of age. During this period, the testicular phosphomonoester/adenosine triphosphate (PM/ATP) ratio changed from 1.70 +/- 0.21 to 0.32 +/- 0.08. There was a significant (P < 0.001) linear correlation between the proportion of haploid cells evaluated by DNA flowcytometry and the PM/ATP ratio evaluated by 31P MRS. 31P MRS is thus a reliable, noninvasive technique for accurately assessing the status of the seminiferous epithelium. PMID- 11409171 TI - Application of a drug delivery system in a novel rat model of chronic hyperendotoxemia. AB - There has not been an ideal reproducible small-animal model of chronic hyperendotoxemia to date. Our drug delivery system (DDS) is a new technology that can deliver a drug conveniently to a target organ at an optional rate. 2 Hydroxyethyl methacrylate (HEMA) was used as a carrier of lipopolysaccharide (LPS), and diethylene glycol and polyethylene glycol dimethacrylates (2G, 4G, 9G) were used as cross-linking agents. A mixed solution of HEMA and di(poly)ethylene glycol dimethacrylate was charged into a glass tube with or without LPS and polymerized by ultraviolet irradiation. This polymer was cut into DDS tablets of the same size with or without LPS. A mixture with HEMA:4G = 1:3 was the most suitable composition to release a constant concentration of LPS. We also developed a novel rat model of chronic hyperendotoxemia. Four DDS tablets, each containing 15 mg LPS, were implanted into the abdominal cavity of rats in the LPS group. The control group was implanted with four DDS tablets without LPS. Plasma levels of LPS in the study group were maintained at more than 2,000 pg/ml for 72 h after implantation. Weight gain was lower and body temperature was higher in the LPS group than in the control group. Plasma levels of inter leukin (IL)-6 in the LPS group were higher than in the control group only during the initial 12 h after implantation of DDS tablets. The white blood cell count at 24 h and platelet counts at 24, 48, and 72 h in the LPS group were lower than those in the control group. These results indicate that chronic hyperendotoxemia was maintained for 72 h by continuous release of LPS from the DDS. Moreover, the intensity of endotoxemia could be varied by varying the number of DDS tablets. It is concluded that our new rat model using LPS-DDS will be applicable and useful as a model of chronic hyperendotoxemia. PMID- 11409172 TI - Do gastrostomies close spontaneously? A review of the fate of gastrostomies following successful renal transplantation in children. AB - Previous published data have shown the benefit of nutritional support delivered via a gastrostomy button (GB) for children on chronic dialysis. The use of the GB is suspended following renal transplantation (RT) in most children and it is usually removed 2-3 months later together with the chronic dialysis catheter when the child is on alternate-day steroids. We reviewed the outcome of gastrostomies following successful RT in children. The gastrostomies were created by an open technique (Stamm) with the child under general anaesthesia, usually at the time of insertion of a chronic dialysis catheter. Growth data and complications of the GB were collected in a prospective registry. Following RT, the GB was removed with the expectation that the tract would close spontaneously. Those in whom a gastrocutaneous fistula persisted underwent formal surgical closure. A total of 18 children have had gastrostomy buttons removed: 11 gastrostomies (61%) closed spontaneously, but 7 (39%) required operative closure at a median of 2 months (range 3 weeks-4 years) post-removal. The need for formal closure was significantly related to the duration that the gastrostomy had been in situ pre transplant (non-parametric statistics, 0.05 > p > 0.01). Although nearly two thirds of gastrostomies in this study closed spontaneously following RT, less than one-half of those that had been in situ for more than 1 year did so. We thus recommend formal closure of all gastrostomies that have been in situ for more than 1 year. This can be done at the same operation as the removal of the chronic dialysis catheter. PMID- 11409173 TI - Surgical treatment of cloacal anomalies. AB - From 1989-1998 14 patients were treated with cloacal anomalies: 5 typical cloacas (PC), 5 posterior cloacas, and 4 cloacal exstrophies (CE); 12 underwent surgery. Four typical cloacas were resolved with posterior sagittal anorectovagino urethroplasty (PSARVUP), whereas in the 5th total urogenital mobilization (TUM) was used. Three PCs were managed with transanorectal TUM and 2 with anterior TUM without opening the anal canal and rectum (without a protective colostomy). Two CEs were treated with atypical procedures. Two patients with CE died without surgery and 2 died after surgery due to complex associated anomalies. During postoperative follow-up of 1-8 years, 5 children had voluntary bowel movements and no soiling while the others had soiling with or without enemas; 1 had stress incontinence; 3 were on clean intermittent catheterization due to neurogenic bladder and were dry. PSARVUP provides a satisfactory result if there is no sacral anomaly. TUM makes this operation easier to perform. In patients with a PC it is sometimes possible using TUM to separate the urinary from the genital tract and remove the accessory urethra without opening the anus and rectum. PMID- 11409174 TI - A novel putative reductase (Cpd1p) and the multidrug exporter Snq2p are involved in resistance to cercosporin and other singlet oxygen-generating photosensitizers in Saccharomyces cerevisiae. AB - Phytopathogenic Cercospora species produce cercosporin, a photoactivated perylenequinone toxin that belongs to a family of photosensitizers which absorb light energy and produce extremely cytotoxic, reactive oxygen species. In this work, we used Saccharomyces cerevisiae as a model system for the identification and cloning of genes whose products mediate cercosporin detoxification. Two genesexpressed in high-copy number vectors conferred cercosporin resistance to an otherwise sensitive strain. One gene codes for Snq2p, a well-characterized multidrug, ABC-type, efflux protein. The other, designated CPD1 (Cercosporin Photosensitizer Detoxification), encodes a novel protein with significant similarity to the FAD-dependent pyridine nucleotide reductases. We showed that over-expression of either of these proteins can also mediate resistance to other singlet oxygen-generating compounds. The involvement of Snq2p and Cpd1p in photosensitizer detoxification reinforces previous observations which suggested that singlet oxygen acts on membrane lipids and that cellular resistance to cercosporin is mediated by a mechanism involving toxin efflux and/or toxin reduction. PMID- 11409175 TI - Diauxic shift-induced stress resistance against hydroperoxides in Saccharomyces cerevisiae is not an adaptive stress response and does not depend on functional mitochondria. AB - Respiring Saccharomyces cerevisiae cells grown on a non-fermentable carbon source are intrinsically more resistant to several stresses, including oxidative stress. The mechanisms leading to increased stress resistance are not yet well understood. Active mitochondria are the major source of intracellular reactive oxygen species (ROS), which could cause the up-regulation of the antioxidant defense systems. We investigated the role of mitochondria in the intrinsic stress resistance against the hydroperoxides H2O2 and tert-butylhydroperoxide 4 h after a shift in carbon source. We found that, independently of functional mitochondria, the yeast acquired the intrinsic resistance of respiring cells against hydroperoxides solely as a response to a change of carbon source in the growth medium. Furthermore, utilizing reporter gene fusion constructs, we monitored the expression of the gamma-glutamylcysteinyl synthetase (encoded by GSH1) and the two superoxide dismutases (encoded by SOD1 and SOD2) during the metabolic transition from fermentation to respiration; and we detected an up regulation of all three genes during the diauxic shift. Overall available data allowed us to propose that the antioxidant system of S. cerevisiae could be considered as a class of genes under glucose/carbon catabolite regulation. This control system is different from the well-known adaptive response to oxidative stress. PMID- 11409176 TI - Phase-specific gene expression in Saccharomyces cerevisiae, using maltose as carbon source under oxygen-limiting conditions. AB - The transcription of ten stress-response genes was investigated under oxygen limiting conditions with maltose and glucose, respectively. Six of these genes (HSP12, HSP26, HSP30, HSP78, HSP82 and HSP104) showed expression only during the stationary phase. HSP12 and HSP104 were transcribed 10 h earlier with maltose than with glucose. Fermentation in wort yielded similar results to the maltose based medium. HSP12, HSP26 and HSP30 were highly expressed. Thus, the HSP26 and the HSP30 promoter can be used for late, phase-specific expression of the desired genes with glucose or maltose as carbon source, and HSP12 with glucose only. MET14, a gene important for sulfite formation, was overexpressed in wort, using the HSP26 promoter during the stationary phase. PMID- 11409177 TI - Cis-acting sites contributing to expression of divergently transcribed DAL1 and DAL4 genes in S. cerevisiae: a word of caution when correlating cis-acting sequences with genome-wide expression analyses. AB - Correlating genome-wide expression profiles with sequence searches of promoter regions is being used as a technique to identify putative binding sites for transacting factors or to refine consensus sequences of those already known. To evaluate the limitations of such an approach in our studies of GATA-mediated transcription in Saccharomyces cerevisiae, we identified the relative contributions made to DAL1 and DAL4 expression by each of five Gln3p-, and/or Gat1p-, and three Dal82p-binding site homologous sequences situated in the 829-bp intergenic region separating these highly related, divergently transcribed genes. Our data suggest that although the correlation of repeated sequences or sequence homologies appearing within promoter regions with expression profiles obtained from genome-wide transcription analyses can provide useful starting points for analyses of cis-acting sites, significant limitations and possibilities for misinterpretation also abound. PMID- 11409178 TI - Fission yeast tor1 functions in response to various stresses including nitrogen starvation, high osmolarity, and high temperature. AB - A target of rapamycin (TOR) protein is a protein kinase that exerts cellular signal transduction to regulate cell growth in response to extracellular nutrient conditions. In the Schizosaccharomyces pombe genome database, there are two genes encoding TOR-related proteins, but their functions have not been analyzed. Here we report that one of the genes, referred to as tor1+, is required for sexual development induced by nitrogen starvation. Ste11 is a key transcription factor for the initiation of sexual development. The expression of ste11+ is normally regulated in tor1- cells; and overexpression of ste11+ hardly rescues the defect in fertility in tor1-. Upon nitrogen starvation, tor1+ cells promote two rounds of the cell cycle to become arrested at the G1 phase before initiation of sexual development. The tor1- cells do not promote such a cell cycle, suggesting that Tor1 is necessary for the response to nitrogen starvation. The tor1- cells show no growth or very slow growth under various stress conditions, including external high pH, high concentrations of salts or sorbitol, and high temperature. These results suggest that Tor1 is necessary for any response to a wide range of stresses. The vegetative growth of tor1- cells is inhibited by rapamycin, although tor1+ cells are resistant to the drug. The tor1- cells are hypersensitive to fluphenazine and cyclosporin A, which specifically inhibit calmodulin and calcineurin, respectively. PMID- 11409179 TI - AoHapB, AoHapC and AoHapE, subunits of the Aspergillus oryzae CCAAT-binding complex, are functionally interchangeable with the corresponding subunits in Aspergillus nidulans. AB - Two genes, AohapB and AohapE, encoding subunits of the Aspergillus oryzae CCAAT binding complex were cloned and sequenced. The polypeptides encoded by AohapB and AohapE were expressed in Escherichia coli and used to reconstitute a DNA-binding complex with recombinant AoHapC. The DNA-binding activity was observed only in the presence of all three subunits, indicating that AoHapB, AoHapE and AoHapC are essential for CCAAT-binding. Furthermore, introduction of the AohapB, AohapC and AohapE genes into the A. nidulans hapB delta, hapC delta and hapE delta strains, respectively, revealed that the A. oryzae Hap subunits are functionally interchangeable with the corresponding subunits in A. nidulans. PMID- 11409180 TI - Primary structure and transcription patterns of RPL36, a ribosomal protein encoding gene of the mycoparasitic fungus, Trichoderma hamatum. AB - We report the isolation and expression profiles of a single-copy gene from the mycoparasitic fungus Trichoderma hamatum encoding a 60 S cytoplasmic ribosomal protein. The gene, named RPL36, was cloned through its nutrient-mediated expression, using mRNA differential screening. Its predicted ORF, interrupted by two introns, encoded a 105-aa polypeptide. The deduced rpL36 protein showed high overall homologies with other L36-type ribosomal proteins isolated from yeast, rat and human. Analysis of the promoter region of RPL36 revealed the presence of two ribosomal protein gene (RPG) boxes and a T-rich region known to be involved in the regulation of most ribosomal protein genes. Expression of RPL36 was tightly regulated by carbon and nitrogen availability. The mRNA levels of this gene decreased upon exposure of the mycelium to different stresses, whereas the addition of cycloheximide resulted in a super-induction. Levels of RPL36 transcripts also increased during mycoparasitic interaction between T. hamatum and Botrytis cinerea. PMID- 11409181 TI - A gene encoding a hydrophobin, fvh1, is specifically expressed after the induction of fruiting in the edible mushroom Flammulina velutipes. AB - A cDNA clone of the gene fvh1 was previously isolated from a preprimordial cDNA library made from the basidiomycete Flammulina velutipes. Sequence analysis showed that fvh1 encoded for a hydrophobin, a small fungal protein usually secreted by filamentous fungi. FVH1 had a highly conserved arrangement of eight cysteine residues, a putative N-terminal signal sequence and a hydropathy pattern characteristics of class I hydrophobin. A genomic fvh1 clone was isolated from a F. velutipes genomic DNA library and sequenced. Several putative promoter elements and three small introns of fvh1 were identified in this clone. Southern blot analysis of genomic DNA showed that fvh1 was a single copy gene. Northern analysis indicated that fvh1 was specifically and abundantly expressed in mycelia after the induction of fruiting and during fruit body initiation. It was not expressed in mycelia before the induction of fruiting or in mature fruit bodies. PMID- 11409182 TI - Mitochondrial control region structure and single site heteroplasmy in the razorbill (Alca torda; Aves). AB - The primary structure of the Alca torda mitochondrial control region was determined and conserved structural features were identified based on sequence comparisons to other bird species. In a population survey using control region analysis, five individuals were found to possess heteroplasmic point mutations at the variable 5' end of the control region. The pattern of variable nucleotide positions among individuals was compared to the distribution of heteroplasmic sites and the heteroplasmic condition was further characterised by a cloning procedure applied to two individuals which harboured one and two heteroplasmic point mutations, respectively. These results are in support of recent evidence that single site heteroplasmy may be more common than previously thought. PMID- 11409183 TI - Mechanisms of weight gain in humans. PMID- 11409184 TI - Effects of oral doxofylline on inflammatory changes and altered cell proliferation in chronic obstructive bronchitis. AB - The effects of oral doxofylline (400 mg BID) on bronchial airway mucosa were investigated in 14 patients with chronic obstructive bronchitis. The eligible patients had to have forced expiratory volume in 1 second > 60% of the predicted value and oxygen partial tension > 55 mm Hg. At the onset and the end of the study, bronchial biopsies were performed via a flexible fiberoptic bronchoscope. Chronic inflammation of the airways was graded according to absence of lesions = 0, involvement of 1-10% = 1, 10-50% = 2 and > 50% = 3. After three months of treatment, 57% of patients in the doxofylline group presented absence of lesions, while the remaining 43% exhibited advanced lesions. In the control group, absence of lesions was observed in 14%, while lesions of grade 1, 2 and 3 were visible in 14%, 29% and 43%, respectively. At histological examination, a significant difference in the degree of structural changes was observed in the doxofylline group between baseline and the end of the study (p < 0.03). In conclusion, doxofylline may induce favorable effects on inflammatory changes and altered cell proliferation of the airway mucosa in patients with chronic obstructive bronchitis. PMID- 11409185 TI - Double-blind, randomized clinical trial of troxerutin-carbazochrome in patients with hemorrhoids. AB - This multicenter, double-blind, randomised study was undertaken to determine the efficacy and safety of a combination of troxerutin 150 mg and carbazochrome 1.5 mg compared to carbazochrome alone in patients with acute uncomplicated hemorrhoids. Patients were administered by the intramuscular route (one ampoule) twice daily for one week. Both subjective and objective efficacy variables significantly improved in the combination drug group only, thus demonstrating the rationale for a combination therapy. Treatments were safe and well tolerated either at a local or systemic level. PMID- 11409186 TI - Blood pressure variations assessed by continuous 24-hour monitoring in menopausal and climacteric women. AB - Changes in the metabolic hormonal balance during the climacteric and menopause, especially surgically induced menopause, increase the risk of acute cerebrocardiovascular complications. This major risk may be linked to changes in blood pressure. In this study we performed twenty-four ambulatory blood pressure monitoring in climacteric (C), menopausal (PM), and surgically induced menopausal women (SM) to determine mean diurnal and nocturnal systodiastolic levels and percentage peaks, as variations in the pressure profile may be linked to organ damage. Our results showed that the entire series presented mainly diastolic increments (mDBP: HPM = 104.4 +/- 5.1; HSM = 106.3 +/- 2.9; HC = 100.2 +/- 3.1), and that this rise was greater in surgically induced menopausal women. In addition, these subjects presented the highest diastolic and systolic pressure peaks (HSM 37/42 versus HPM 35/36 and HC 29/31) also during the night (nocturnal peak: HSM 15/19 versus HPM 10/12 and HC 5/15). Non dippers seem more exposed to cerebrocardiovascular disease. Our results revealed that climacteric patients affected by arterial hypertension (mSBP = 162.2 +/- 4.1; mDBP = 100.2 +/- 3.1; 24 h systolic peak % = 24, diastolic peak % = 24) during the climacteric presented the same levels as observed in conclaimed menopause (mSBP 165.2 +/- 5.5; mDBP = 104.2 +/- 5.1; 24 h systolic peak % = 28, diastolic peak % = 29). Therefore, 24 h blood pressure monitoring is able to show that the pressure changes in hypertensive climacteric and menopausal women and could detect women who are at a greater risk of organ damage. PMID- 11409187 TI - Hyperresponsiveness of airway muscle to acetylcholine in asthmatic and non asthmatic subjects. AB - BACKGROUND: Despite a long-standing recognition, the bronchial hyperresponsiveness (BHR) of airway smooth muscle, in asthmatic and non-asthmatic subjects is still under investigation. OBJECTIVE: In order to better understand the mechanism regulating the airway function, some results obtained with cellular physiology techniques are compared, for some non-asthmatic without BHR subjects and other subjects suffering from real bronchial asthma. SUBJECTS: Eighteen subjects without chronic or acute bronchopathy and seven patients with a typical history of bronchial asthma were considered. All of them were non-smokers, with a wide range of in vivo hyperresponsiveness to Acetylcholine (ACh). Owing to neoplastic lesions, all patients underwent a thoracic surgery. MEASUREMENTS AND RESULTS: In vitro hyperresponsiveness of airway-smooth-muscle chains was assessed with isotonic contractions to incremental doses of ACh, in accordance with a non cumulative dose-response technique. Regression between muscle contractions and ACh doses was studied for both groups, and it turned out to be linear in both cases. The slopes and elevations of the regression lines of the two groups were statically identical. There was no correlation between in vivo hyperresponsiveness (as PD20-ACh-FEV1) and in vitro responses (as EC50), for both asthmatic and non-asthmatic group. CONCLUSIONS: (i) in vitro, the same BHR of smooth-muscle resulted, for both asthmatic and non-asthmatic subjects, (ii) in vivo, a different BHR of the former group resulted. This can be explained with the peculiarity of the micro-environmental surrounding the bronchial tissue, not with an intrinsic defect of the isolated bronchial tissue. PMID- 11409188 TI - Therapeutic approach of carcinoid tumours of the lung. AB - In the carcinoid tumours of the bronchopulmonary tract surgical resection is still the primary goal. Many problems are, however, unclear: the extent of resection, formal lymph node dissection or not, the role of Video-Assisted Thoracic Surgery (VATS) and of the multidisciplinary approach. In the Department of Surgical Sciences and Applied Medical Technologies, "La Sapienza", Rome's University, from 1969 to 1994, we observed 18 patients with carcinoid tumours of the lung: 13 typical carcinoid (TC) and 5 atypical carcinoid (AC). In our series, the choice of therapeutic procedure was made on the basis of histological criteria and TNM classification. We performed 3 conservative and 10 extensive resections on typical carcinoid and 5 extensive resections on atypical carcinoid tumours. In our series VATS played a minor therapeutic role. Formal lymph node dissection was carried out on all our patients except in the cases of those with typical carcinoid tumours without enlarged hilar and mediastinal lymph nodes. The efficacy of adjuvant chemotherapy in carcinoid tumours treatment is controversial and will be confirmed by further trials. In bronchial carcinoid tumours the long term prognosis is excellent. In our series the ten-year survival rate is 77 per cent in typical carcinoid and 40 per cent in atypical carcinoid cases. PMID- 11409189 TI - Giant hepatomegaly and portal hypertension in an elderly patient with primary liver amyloidosis: an uncommon clinical occurrence. AB - Primary systemic amyloidosis (AL) is an uncommon disease characterized by the extracellular deposition of a protein with a beta-fibrillar structure, consisting of monoclonal immunoglobulin light chains, lambda or kappa (ratio of lambda to kappa, 3:1). In systemic amyloidosis liver involvement is frequent but it rarely has clinical importance. The massive and localized liver deposition of amyloid, characterized by marked hepatomegaly and portal hypertension without hepato cellular failure and by a severe prognosis, without systemic involvement, is less frequent. The authors describe an unusual case of primary hepatic amyloidosis with giant hepatomegaly, intrahepatic cholestasis, portal hypertension and splenomegaly, occurred in an elderly patient. PMID- 11409190 TI - Comparative bioavailability of a new formulation of fluoxetine drops. AB - Fluoxetine drops and marketed fluoxetine capsules had quite the same Cmax (50.25 +/- 4.43 vs 47.55 +/- 5.29 ng/ml), but significantly different AUC0-t (717.27 +/- 71.29 vs 644.91 +/- 78.91 ng/ml/h). Furthermore the drops were characterised by a very early Tmax (4.61 +/- 0.85 hours) with a highly significant difference in comparison to the capsules reference compound (6.72 +/- 1.23 hours). After log transformation 90% C.I. for Cmax, AUC 0-t and AUC0-. were 1.05, 1.11, 0.82 respectively. The two products, therefore, cannot be considered bioequivalent. Our results demonstrate that fluoxetine drops and capsules significantly differ for their pharmacokinetics, with an earlier Tmax and a higher AUC0-t after the administration of the drops preparation. PMID- 11409191 TI - Preliminary risk assessment of the wet landscape option for reclamation of oil sands mine tailings: bioassays with mature fine tailings pore water. AB - Chemical and biological assays have been carried out on the "pore water" that results from the settling of the tailings that accompany bitumen recovery from the Athabasca oil sands. Examination of the nonacidic extracts of pore water by gas chromatography-mass spectroscopy allowed the identification of numerous two- to three-ring polycyclic aromatic compounds (PACs), to a total concentration of 2.6 micrograms/L of pore water. The PACs were biodegraded by microflora naturally present in the pore water. Acute toxicity was associated principally with the acidic fraction (naphthenic acids) of pore water extracts according to the Microtox assay; other work has shown that acute toxicity dissipates fairly rapidly. Both individual PACs and concentrated pore water extracts showed minimal levels of binding to the rat Ah receptor and induced minimal ethoxyresorufin-O deethylase activity in primary rat hepatocytes, showing an insignificant risk of inducing monooxygenase activity. Taken together with previous work showing negligible mutagenic activity of these extracts, we conclude that it should be possible to develop tailing slurries into biologically productive artificial lakes. PMID- 11409192 TI - Transcriptional activation of stress genes and cytotoxicity in human liver carcinoma cells (HepG2) exposed to 2,4,6-trinitrotoluene, 2,4-dinitrotoluene, and 2,6-dinitrotoluene. AB - The CAT-Tox (L) assay has recently been developed and validated for detecting and quantifying the specific molecular mechanisms that underlie toxicity of various xenobotic chemicals. We performed this assay to measure the transcriptional responses associated with 2,4,6-trinitrotoluene (TNT) and 2 of its byproducts [2,4 and 2,6-dinitotoluenes (DNTs)] to 13 different recombinant cell lines generated from human liver carcinoma cells (HepG2) by creating stable transfectants of mammalian promoter chloramphenicol acetyltransferase (CAT) gene fusions. Cytoxicity test with the parental HepG2 cells, using the MTT [3-(4,5 dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide]-based assay for cell viability, yielded LC50 values of 105 +/- 6 mg/mL for TNT in 1% dimethyl sulfoxide (DMSO), and > 300 mg/mL for DNTs, upon 48 h of exposure. TNT appeared to be more toxic than 2,4-DNT, which also showed a higher toxicity compared to 2,6-DNT. Of the 13 recombinant constructs evaluated, 8 (CYP 1A1, GST Ya, XRE, HMTIIA, c-fos, HSP70, GADD153, and GADD45), 5 (c-fos, HSP70, GADD153, GADD45, and GRP78), and none showed inductions to significant levels (p < 0.05), for TNT, 2,4 DNT, and 2,6-DNT, respectively. For most constructs, the induction of stress genes was concentration-dependent. These results show the potential for TNT and 2,4-DNT to cause protein damage and/or perturbations of protein biosynthesis (HSP70 and GRP78), alterations in DNA sequence or its helical structure (c-fos, GADD153, GADD45), and the potential involvement of TNT in the biotransformation process (CYP 1A1, GST Ya, XRE), and in the toxicokinetics of metal ions (HMTIIA). Within the range of concentrations tested (0-300 mg TNT or DNT/mL in 1% DMSO), no significant inductions (p > 0.05) of NFKBRE, p53RE, CRE, and RARE were found. PMID- 11409193 TI - Toxicity tests to assess pollutants removal during wastewater treatment and the quality of receiving waters in Argentina. AB - In Argentina, legislation to control adverse impacts of effluent discharges and the quality of receiving waters is scant and relies mainly on the physicochemical characteristics of the effluents and receiving waters. Objectives of this study were to use standardized acute toxicity tests to assess treatment of petrochemical industry effluents and the toxicity of various treated industrial effluents in the Buenos Aires metropolitan area and their receiving waters. Tests for the first objective used Daphnia magna and Ceriodaphnia dubia; those for the second used D. magna, Spirillum volutans, and Scenedesmus spinosus. Chemical analyses demonstrated that the removal of aromatic hydrocarbon compounds (benzene, toluene, ethylbenzene, xylene, styrene, and naphthalene) from the petrochemical effluents ranged between 77 and 93%, but toxicity removal was significantly lower: untreated effluents were very toxic and treated effluents were very toxic to toxic [acute toxicity units (TUa) > 3]. Physicochemical parameters measured according to current Argentinian regulations indicated that industrial effluents (e.g., from textile and paper industries) were within established guidelines, but 25% of the samples were moderately to highly toxic (TUa > 1.33). However, for the receiving waters, toxicity tests were moderate to very toxic. The results show the need of including tests for toxicity of discharged effluents, and their effects on receiving waters of Argentina, especially for regulatory purposes. PMID- 11409194 TI - Apoptotic effect of cyanobacterial extract on rat hepatocytes and human lymphocytes. AB - Toxic cyanobacterial blooms are an increasing problem in Poland. The production of cyanobacterial toxins and their presence in drinking and recreational waters represent a growing danger to human and animal health. This is connected with the increase of cyanobacterial biomass caused by excessive eutrophication of the water ecosystem. There is evidence that cyanobacterial hepatotoxins can act as a potent promoter of primary liver cancer. The apoptotic effect of microcystins in Polish cyanobacterial bloom samples on rat hepatocytes and human lymphocytes was observed using light and fluorescence microscopy, flow cytometry, and electrophoretic analysis. The incubation time needed to observe the first morphological apoptotic changes in hepatocytes was approximately 30 min; however, the characteristic biochemical changes in DNA were not observed even after 120 min. In lymphocyte cultures the morphological changes characteristic for apoptosis were observed after 24 h of incubation and a 48-h incubation was found to be optimal for analysis of internucleosomal DNA fragmentation, which is one of the main biochemical hallmarks of programmed cell death. These cells are an easily isolated and inexpensive material for medical diagnostics. Therefore the apoptotic changes, together with the clastogenic effect seen in lymphocyte cultures, are proposed as a future analytical method for these toxins. PMID- 11409195 TI - Use of imposex (pseudohermaphroditism) as indicator of the occurrence of organotin compounds in Portuguese coastal waters--Sado and Mira estuaries. AB - Organotin compounds, including tributyltin (TBT), are a class of the most toxic xenobiotics occurring in aquatic systems. High concentration levels in waters and sediments are mainly due to their extensive use as biocides and high persistence when present in sediments under anaerobic conditions. Toxicity studies have revealed the acute effects of TBT for aquatic organisms at concentrations as low as 1 microgram/L, and the induction of imposex at levels below 0.5 ng/L TBT (as Sn). At 20 ng/L TBT (as Sn) causes sterility and is followed with the disappearance of the most sensitive neogastropods on a given shore. Imposex is the most sensitive response of all known pathological conditions for nontarget organisms following an exposure to tributyltin. In this study results are discussed that were obtained from two monitoring sites with different anthropogenic background using imposex monitoring as an indicator of TBT concentrations, as well as chemical analysis of tissue of Hinia (= Nassarius) reticulata (L.) (Gastropoda). PMID- 11409196 TI - A colorimetric protein phosphatase inhibition assay for the determination of cyanobacterial peptide hepatotoxins based on the dephosphorylation of phosvitin by recombinant protein phosphatase 1. AB - A colorimetric protein phosphatase inhibition assay based on the dephosphorylation of phosvitin by recombinant protein phosphatase 1 was developed for analysis of waters for cyanobacterial hepatotoxins. The phosphate released in the assay was determined using a malachite green reagent. Good agreement with toxin concentrations determined by HPLC was obtained. The assay was capable of determining these toxins at concentrations around 1 microgram/L with high precision and without sample concentration. This is of considerable benefit as the World Health Organisation specifies a provisional guideline of 1 microgram/L for microcystin-LR. There was evidence, however, that the sample matrix might affect quantification, leading to false positive results. Thus the assay should be viewed as a screening procedure, and confirmatory analyses by an alternative procedure should be carried out for positive results. Further work is required to resolve the question of matrix interferences if phosphatase inhibition assays are used directly for measuring toxin levels in water, especially if this information is used to check compliance with water quality guidelines. PMID- 11409197 TI - Heat shock protein 70 levels in rainbow trout primary epidermal cultures in response to 2,4-dichloroaniline exposure: a novel in vitro aquatic toxicity marker. AB - The aim of this work was to investigate the use of the heat shock protein, HSP 70, as a sublethal measurement of ecotoxicity and to identify if the amount of HSP 70 synthesized is proportional to the chemical stress applied. This was achieved by quantifying the HSP 70 levels in primary cultured rainbow trout, Oncorhynchus mykiss (R.), skin epidermal cells in response to 2,4-dichloroaniline (2,4-DCA) exposure. The cellular stress response protects organisms from damage resulting from exposure to a wide variety of stressors including xenobiotics. The use of a HSP 70 polyclonal antibody on rainbow trout primary epidermal skin cultures exposed to 2,4-DCA was investigated as a possible biomarker for environmental stress using an immunocytochemical approach. The epidermis is highly susceptible, as it is the interface between the fish and its aquatic environment. In this study we have developed a simple in vitro system for aquatic toxicity risk assessment. A method for the quantification of heat shock (stress) protein levels by immunocytochemistry is described. The antibody dilution range enabled the detection and quantification of only the inducible HSP 70 fraction. A 1:2000 dilution was decided upon. This assay was effective in detecting and quantifying the induced HSP 70. There was a direct toxicant concentration dependent increase in the levels of the cellular stress protein in the primary epidermal cultures. Enhanced localization of HSP 70 in the nuclei of the epidermal cells was observed following exposure to 2,4-DCA. This work indicated the possibility of using heat shock protein induction and subsequent quantification as a sensitive system for aquatic toxicity risk assessment. PMID- 11409198 TI - Occurrence of compounds estrogenic to freshwater mussels in surface waters in an urban area. AB - Estrogens play a major role in the sexual differentiation, gonad development, and oocyte growth of most oviparous organisms. They also stimulate vitellogenesis, the formation of high-density glycolipophosphoprotein that serves as an energy source for the developing embryo. Surface waters from the St. Lawrence River, obtained in the vicinity of an urban area (Montreal, Quebec, Canada), were studied with respect to their estrogenic potential to the freshwater mussel Elliptio complanata. Estrogenicity was measured in water extracts by means of a competitive assay of estradiol binding to cytosolic proteins and by the vitellin inducing ability of mussel hemolymph following direct extract injection. Surface water samples drawn downstream of a municipal outfall plume and in a river draining a large farming and agricultural area had high levels of total and fecal coliform bacteria. High levels of estrogen competitors were also found and were able to induce vitellins in injected mussels. Moreover, the estrogen-competing potential of the extracts was found to be significantly correlated with total and fecal coliform bacteria (R = 0.9, p < 0.01) and with the levels of vitellins in the hemolymph (R = 0.62, p = 0.03). The results indicate that water samples drawn from within the municipal effluent plume and from a river draining an agricultural area are estrogenic to freshwater mussels. Thus, the environmental inputs of estrogens are likely to be associated with human sewage and pesticide products. PMID- 11409199 TI - Effect of sulfur dioxide and particulate pollutants on bronchitis in children--a risk analysis. AB - The general morbidity is being influenced to a great extent by diseases of the respiratory tract. Since their incidence and prevalence have been increasing, the identification of causal factors, especially of environmental origin, is of importance, not just in view of implementing preventive control strategies. Primary irritative gaseous [like sulfur dioxide (SO2)] as well as particulate pollutants (like TSP) can be regarded as causal constituents. Using the opportunity of changing levels of ambient air pollution in East Germany since 1989, the impact of SO2 and TSP on bronchitis was investigated over the last 10 years as part of several intervention studies in a locally defined homogenous population, children. The data suggest a significant association between SO2 and the prevalence of bronchitis in children but not for TSP. Considering the findings of other studies with respect to air pollution and the epidemiology of adverse health effects (especially respiratory disease), these results would amend the hypothesis put forward in a review of the literature as TSP < ultrafine particles (SO4(2-)) = SO2. Although these data show a significant association with only SO2, this does not imply that ultrafine particles, such as SO4(2-), do not contribute to the association with the observed adverse health effects. Nevertheless, the findings suggest that TSP seems less likely to be a predictor in the association with respiratory diseases, particularly not in the presence of high SO2. PMID- 11409200 TI - Effects of a pre-incubation period on the photoinduced toxicity of polycyclic aromatic hydrocarbons to the luminescent bacterium Vibrio fischeri. AB - Irradiation of polycyclic aromatic hydrocarbons (PAHs) in aqueous solution with simulated solar radiation (SSR; a light source with a visible light: UV-A:UV-B ratio similar to that of sunlight) can greatly enhance their toxicity. Two microbial toxicity tests with Vibrio fischeri were used to investigate the effect of composition of the growth medium and pre-incubation on the photoinduced toxicity of PAHs. The assays were a short-term test (15 min) and long-term test (18 h). Both assays were carried out in SSR and darkness to examine for photoinduced toxicity of PAHs. For the short-term toxicity assay, inhibition of bacterial luminescence was measured. For the long-term toxicity assay, both inhibition of bacterial luminescence and inhibition of growth were recorded. To broaden this test, V. fischeri cells were pre-incubated with PAHs in medium without a carbon source (minimal medium) for 8 h to facilitate assimilation and photooxidation of the contaminants, and to prevent bacterial growth at the outset of the assay. V. fischeri was more sensitive in minimal medium than in complex medium in both the short- and long-term toxicity assays. Moreover, in the long term assay, SSR greatly increased toxicity, especially if there was a pre incubation period in minimal medium. This indicates that both assimilation and photooxidation of PAHs are important to their toxicity to V. fischeri. PMID- 11409201 TI - Hot products 2001. PMID- 11409202 TI - Exposed. Signs, symptoms & EMS management of nerve-agent poisoning. PMID- 11409203 TI - MCI (mass casualty incident) success story. How an EMS system improved its MCI response strategy through planning & practice. PMID- 11409204 TI - MCI (mass casualty incident) resource guide 2001. PMID- 11409205 TI - Designer drugs. Assess & manage patients intoxicated with ecstasy, GHB or rohypnol--the three most commonly abused designer drugs. PMID- 11409206 TI - Foley prices steady, for now. PMID- 11409207 TI - Hospitals awaiting arrival of e-commerce standards. PMID- 11409208 TI - In sterile processing, little things count. PMID- 11409209 TI - The AIDS battle fatigue syndrome: what's challenging medication adherence in HIV? PMID- 11409210 TI - A new technology for detecting HIV: finding the productivity infected cell. PMID- 11409211 TI - Genetic testing provides challenge to insurers. PMID- 11409212 TI - The perils of nonprospectively planned research. Part 2: Guidelines for reviewing the results of sample-based research. PMID- 11409213 TI - Determination of chlorinated solvents in exhaled air, urine, and blood of subjects exposed in the workplace using SPME and GC-MS. PMID- 11409214 TI - Interactive on-line product demonstrations: watch a product work. PMID- 11409215 TI - Ergonomic considerations in the development of a class II, type A/B3 biological safety cabinet. PMID- 11409216 TI - Carbohydrate-deficient transferrin: what have we learned in the last decade? PMID- 11409217 TI - Talking effect and "white coat" effect in hypertensive patients: physical effort or emotional content? AB - Talking has been shown to increase blood pressure instantaneously in hypertensive patients and to contribute to the white coat effect. The effects of talking were compared with those of counting aloud in 64 patients with essential hypertension who were randomly assigned to a period of stress talking and a period of counting aloud (active periods), alternating with three periods of silence (control). The same monitor was used for office measurements and 24-hour ambulatory blood pressure analysis. Systolic/diastolic blood pressures increased significantly more during talking (163/110 mmHg) than during counting aloud (152/102 mmHg, both p < .0001) in both treated and untreated patients and in sustained and clinical hypertension. Talking had a residual effect on systolic blood pressure that lasted 5.8 +/- 0.1 minutes. The emotional content seemed to be the only cause of the talking effect. Its instantaneous and residual effects on blood pressure and heart rate should be considered when measuring these variables. PMID- 11409218 TI - Effects of gender and marital status on somatic symptoms of patients attending a mind/body medicine clinic. AB - To clarify the mechanisms of gender-related mind/body relationships, the authors analyzed the characteristics of 1,132 outpatients (848 women and 284 men) attending a mind/body medicine clinic. At entry in the program, the patients completed the Medical Symptom Checklist, Symptom Checklist-90 revised (SCL-90R), and Stress Perception Scale. Women reported 9 out of 12 symptoms (fatigue, insomnia, headache, back pain, joint or limb pain, palpitations, constipation, nausea, and dizziness) more frequently than the men did. Being a woman was a predictor of the total number of somatic symptoms endorsed. SCL-90R somatization scores were significantly higher in nonmarried women than in married women. Perceived stress ratings of family and health were higher in women than in men, despite the lower degree of perceived stress concerning work. Women, especially nonmarried women, were more likely to report somatic discomfort. Gender appears to be an important factor in relation to the report of somatic symptoms in stress related conditions. PMID- 11409219 TI - Somatization and symptom reduction through a behavioral medicine intervention in a mind/body medicine clinic. AB - The authors assessed data from 1,148 outpatients in a 10-week medical symptom reduction program to determine the effectiveness of a behavioral medicine intervention among somatizing patients. The program included instruction in the relaxation response, cognitive restructuring, nutrition, and exercise. Before and after the intervention, the patients were evaluated on the Symptom Checklist-90 Revised (SCL-90R), the Medical Symptom Checklist, and the Stress Perception Scale. They were divided into high- and low-somatizing groups on the basis of the pretreatment SCL-90R somatization scale. At the end of the program, physical and psychological symptoms on the Medical Symptom Checklist and the SCL-90R were significantly reduced in both groups, with the reductions greater in the high somatizing group. Improvements in stress perception were about the same in both groups, but the absence of an untreated control group precluded estimates of how much the improvements resulted from the behavioral medicine intervention and how much from natural healing over time. PMID- 11409220 TI - Depression and education as predicting factors for completion of a behavioral medicine intervention in a mind/body medicine clinic. AB - The authors compared characteristics of 1,012 outpatients completing a 10-week behavioral medicine intervention with 300 outpatients who dropped out. They administered the Symptom Checklist-90 Revised (SCL-90R) before and after the program. Patients who completed the treatment, compared with dropouts, tended to be more highly educated, married, and gainfully employed. Their pretreatment scores on the SCL-90R were significantly lower than those of the dropouts on somatization, depression, and obsessive-compulsive scales and on the global severity index. Multiple logistic regression analysis indicated that lower depression and higher education marked the group who completed the intervention in contrast to the dropouts. After the intervention, all of the SCL-90R scores were significantly lower among patients who completed the treatment. Pre- to postintervention score changes were not significantly associated with the number of sessions attended. The findings suggest that the intervention had salutary effects in patients with mind/body distress and that its effectiveness was not diminished by a few absences. Depressed or less educated patients might benefit from preparatory interventions or from a modified approach to their treatment. PMID- 11409221 TI - Relationship between ventilatory expired gas and cardiac parameters during symptom-limited exercise testing in patients with heart failure. AB - PURPOSE: This study investigates the relationship between ventilatory expired gas and cardiac parameters measured during exercise testing in patients with heart failure. METHODS: Twenty-five subjects (12 male, 13 female) diagnosed with compensated heart failure underwent symptom-limited exercise testing with ventilatory expired gas analysis. Metabolic and cardiac measures of interest were collected during testing. RESULTS: Mean peak oxygen consumption (VO2), minute ventilation/carbon dioxide production (VE/VCO2) slope, percentage of age predicted maximal heart rate achieved during exercise testing (%APMHR), and peak respiratory exchange ratio were 14.7 +/- 4.7 mL O2/kg/min-1, 33.8 +/- 9.8, 76% +/ 15%, and 1.1 +/- 0.11, respectively. The VE/VCO2 slope was significantly correlated with the following: %APMHR (r = -0.81, P < 0.001), peak VO2 (r = 0.83, P < 0.001), VO2 at ventilatory threshold (r = -0.70, P < 0.001), and the dead space to tidal volume ratio (VD/Vt) (r = 0.65, P < 0.001). The ability of peak VO2 and %APMHR to predict the VE/VCO2 slope was significant (r = 0.86, r2 = 0.72, P < 0.0001). CONCLUSION: This study demonstrates the importance of analyzing multiple exercise test parameters, including metabolic measures, in patients with heart failure. PMID- 11409222 TI - Proportional assist ventilation may improve exercise performance in severe chronic obstructive pulmonary disease. AB - PURPOSE: Exercise tolerance is impaired in chronic obstructive pulmonary disease (COPD), in part because of a reduction in ventilatory capacity and excessive dyspnea experienced. The authors reasoned that proportional assist ventilation (PAV), a ventilator mode in which the level of support varies proportionately with patient effort, could be used during exercise to assist ventilation. The purpose of this study was to evaluate the efficacy of PAV to improve exercise endurance and related physiologic parameters in COPD. METHODS: In 8 patients (age = 62.8 years mean, +/- 6.9 standard deviation) with severe COPD (forced expiratory volume in 1 second = 0.70 +/- 0.21 L) flow, volume, dyspnea, leg fatigue, arterial blood gases, and gas exchange were measured during constant workrate exercise (37 +/- 18 watts; i.e., 80% previously determined maximum oxygen consumption). Crossover exercise trials were performed in random order: while spontaneously breathing through the experimental circuit without assistance (control trial) and with PAV (using 9.8 +/- 2.1 cm H2O/L and 3.3 +/- 1.0 cm H2O/L/sec of volume assist and flow assist, respectively). RESULTS: The application of PAV during exercise was well tolerated by each subject. Compared with the control measurement at equivalent time during exercise, PAV improved breathing pattern and arterial blood gases while dyspnea was reduced. Consequently, there was a significant increase in exercise duration with PAV (323 +/- 245 seconds during the control trial compared with 507 +/- 334 seconds with PAV, P = 0.02). CONCLUSIONS: Proportional assist ventilation can improve performance during constant workrate exercise in severe COPD. PMID- 11409223 TI - Activity monitors can detect brisk walking in patients with chronic obstructive pulmonary disease. AB - PURPOSE: Brisk walking forms the foundation of aerobic training regimes within a pulmonary rehabilitation service for patients with chronic obstructive pulmonary disease (COPD). Repeated bouts of aerobic training are encouraged in the domestic environment. This study was designed to examine the discriminatory properties of an activity monitor, to assess if brisk walking can be identified clearly from all other activities of daily living. Secondly, the authors examined the overall level of activity generated by patients with COPD compared with a healthy control population. METHODS: Eleven patients with COPD and nine age-matched healthy controls were recruited. All subjects were required to wear an activity monitor for 48 hours. Subjects simultaneously recorded their activity (grouped into 10 categories) to the nearest minute. RESULTS: The data indicated that the monitors can detect brisk walking as a distinct activity in patients and a control population. The healthy individuals achieve a significantly higher level of activity than patients, but that the duration of this high intensity activity (i.e., brisk walking) is significantly lower than the patient group. Total activity was not correlated with age, sex, or forced expiratory volume in 1 second. CONCLUSION: Activity monitors have the capacity to discriminate brisk walking from other domestic activities and can be used to identify low levels of domestic activity in the patient population. PMID- 11409224 TI - The National Lung Health Education Program. A new healthcare initiative for America. PMID- 11409225 TI - Effects of home versus supervised exercise for patients with intermittent claudication. AB - PURPOSE: This study was performed to test the efficacy of a supervised, hospital based exercise program compared with a home-based exercise program involving minimal supervision, for both walking ability and quality of life measures in patients with exercise-limiting intermittent leg claudication. METHODS: Twenty one patients were assigned randomly to 12 weeks of supervised exercise or to a home-based exercise group. After 12 weeks the participants in the supervised group transitioned to a home-based program. Both groups were then reevaluated at the end of 24 weeks. The initial claudication distance (ICD) and absolute claudication distance (ACD) on progressive treadmill exercise was measured at baseline, 12 weeks, and 24 weeks. Additionally, self-reported quality of life status was evaluated using the MOS SF-36 questionnaire. RESULTS: Each group improved (P < 0.01) ACD from baseline to 12 weeks, which was sustained at the 24 week follow-up. Both groups experienced similar long-term improvements (P < 0.05) in ACD (521.5 +/- 253.4 meters to 741.9 +/- 365.6 meters for the supervised group, 532.2 +/- 263.5 meters to 715.0 +/- 394.4 meters in the home group, P not significant, between groups). The supervised group experienced a greater improvement (P < 0.01) in the ICD after 12 weeks than the home group but not at 24 weeks. The on-site group also experienced significant improvements in ICD after 24 weeks (P < 0.05). Neither group manifested an improvement in self reported physical function or mental health as assessed by the MOS SF-36. CONCLUSION: A structured exercise program was more effective in improving the ICD over a 24-week period than a less formal, home-based program. However, if patients are screened properly and receive adequate instruction, a home-based program can be a safe, low-cost alternative providing similar long-term (24 weeks) exercise benefits in ACD. PMID- 11409226 TI - Accuracy of estimating exercise prescription intensity in patients with left ventricular systolic dysfunction. AB - PURPOSE: Exercise prescription in patients with left ventricular systolic dysfunction (LVSD) is difficult. Exercising beyond ventilatory threshold (VT) can have negative physiologic effects; therefore, exercise prescribed above VT may be detrimental. A majority of cardiac rehabilitation programs use the Karvonen/heart rate reserve (HRR) method, rating of perceived exertion (RPE), and/or a percentage of oxygen consumption to prescribe exercise intensity. The purpose of this study was to determine if these methods correlate with an exercise intensity below VT in LVSD patients. METHODS: The authors studied 52 patients (37 males, 15 females; age 52 +/- 13 years; left ventricular ejection fraction 27% +/- 8%) who underwent a symptom-limited cardiopulmonary exercise test and reached VT to determine functional capacity and exercise prescription. RESULTS: Peak heart rate (HR) as well as HRR derived minimum (60%), midpoint (70%), and maximum (80%) HR were highly correlated (P < 0.001) with HR at VT. Using these three different HR cutoff formulas from HRR, 15% to 62% of patients were prescribed exercise outside the range of VT-HR +/- 10%. The midpoint (70% HRR) best predicted exercise HR in the VT-HR +/- 10% range (73% of patients). Mean oxygen consumption at VT was 83 +/- 9% of peak oxygen consumption. There was no correlation (P < 0.16) between RPE and VT. CONCLUSIONS: The Karvonen/HRR method failed to estimate HR-VT +/- 10% in a large percentage of patients with LVSD. There was no correlation between RPE and VT. Based on these data, exercise training intensity should ideally be prescribed based on the HR identified at VT using cardiopulmonary exercise testing in patients with LVSD. PMID- 11409227 TI - How hard should we exercise the failing human heart? PMID- 11409228 TI - A comparison of ambulatory oxygen consumption during circuit training and aerobic exercise in patients with chronic heart failure. AB - PURPOSE: This study compared the exercise intensity of a combined aerobic and resistance exercise circuit training session with the exercise intensity of continuous aerobic exercise in patients with chronic heart failure (CHF). METHODS: Peak oxygen consumption (VO2peak) and muscular strength (1 repetition maximum) were assessed in six CHF patients (age 62 +/- 3 years). Heart rate, rate of perceived exertion (RPE), blood pressures, ambulatory oxygen consumption (VO2), and ventilatory data were measured during two types of exercise: continuous cycling on a bicycle ergometer (aerobic [AER] session) and combined AER and resistance exercise (circuit training [CIR] session). RESULTS: There were no significant differences in VO2, RPE, heart rate, or hemodynamic responses (rate pressure product, diastolic blood pressure, or mean arterial pressure) during exercise, between the two sessions. Systolic blood pressure was significantly lower during CIR (P < 0.05). Minute ventilation and tidal volume were significantly higher (P < 0.0001 and P < 0.01, respectively) and respiratory frequency significantly lower (P < 0.005) during CIR. During CIR, RPE significantly correlated with VO2 (P < 0.01), whereas heart rate did not. Conversely, during the AER session HR correlated with VO2 (P < 0.01), but RPE did not. CONCLUSIONS: Circuit training is a well-tolerated form of exercise training for CHF patients that is associated with similar oxygen and hemodynamic demand to aerobic exercise. Results suggest that RPE may be a better method of prescribing and monitoring exercise intensity during CIR, with heart rate the preferred measure of intensity during aerobic exercise. PMID- 11409229 TI - Immeasurable, invisible, or subtle--advantages make a difference. PMID- 11409230 TI - Gloves and glutaraldehyde. PMID- 11409232 TI - Operating room nurse managers--competence and beyond. AB - Operating room nurse managers supervise and direct a complex, highly technical, and expensive organization. To be successful, they must possess certain competencies or skills that have not been studied extensively. A replication of one study was conducted to determine which competencies characterize OR nurse managers who are considered competent and superior. The randomly selected, national sample comprised OR nurse managers who completed an Operating Room Nurse Manager Questionnaire. Analysis of 120 useable questionnaires revealed that human and leadership competencies were valued most and conceptual and technical competencies were valued least in OR nurse managers. PMID- 11409233 TI - Quantifying and reducing the risk of bloodborne pathogen exposure. AB - The risk of becoming infected with bloodborne pathogens (e.g., hepatitis B, hepatitis C, HIV) during surgery is real. The degree of risk for perioperative personnel is related to factors that include participating in large numbers of surgical procedures each year; the nature of perioperative work (e.g., use of different types of sharp instruments): exposure to large amounts of blood and body fluids; the prevalence of bloodborne pathogens in the surgical population; the variation in different organisms' ability to be transmitted; the existence of vaccines and the level of vaccination; the availability of postexposure treatment; and the consequences of acquiring the disease. Controlling risks to perioperative personnel can be accomplished by using the Occupational Safety and Health Administration's three methods of control--redesigning surgical equipment and procedures, changing work practices, and enhancing the personal protection equipment of perioperative personnel. PMID- 11409234 TI - Grounded theory as a qualitative research methodology. AB - Grounded theory as a qualitative research approach provides nurses with strategies to build theories in areas previously unexplored or under explored. Although similar to other inductive processes, this methodology differs in that it emerges from the discipline of sociology. In addition, strategies for data analysis differ according to each methodologist. As with other qualitative methods, the researcher must link philosophy, methodology, and research method for rigorous and credible scholarship. PMID- 11409235 TI - Disinfecting endoscopes immediately before the first patient of the day. PMID- 11409236 TI - The digital extensor mechanism of the hand. AB - In summary, there is a complex interplay between the extrinsic and intrinsic components of the digital extensor mechanism. Numerous examples of dynamic anatomy and pathology of the extensor mechanism in common clinical settings have been described. Treatment principles have been outlined. This review should provide the orthopaedic surgeon with a framework for management of common hand problems. PMID- 11409237 TI - Congenital anomalies of the thumb. A brief review. AB - Congenital anomalies of the thumb are rare and challenging conditions. There are many possible ways of addressing these various disorders, with potential to improve the function and appearance of the hand. PMID- 11409238 TI - Rotatory subluxation of the scaphoid. AB - Rotatory subluxation of the scaphoid remains a challenging problem; a thorough understanding of the basic anatomy, particularly the ligament complexes of the wrist, as well as wrist kinematics are required for the proper diagnosis and treatment. Our understanding of rotatory subluxation of the scaphoid has improved over the years and continues to expand. PMID- 11409239 TI - Cervical spine injuries in football. AB - The game of football, as it is played today, poses serious risk of injury for players of all ages. Injury may occur to any structure of the spinal column, including its bony, ligamentous and soft tissue components. The majority of cervical spine injuries occurring in football are self limited, and a full recovery can be expected. While these injuries are relatively uncommon, cervical spine injuries represent a significant proportion of athletic injuries that can produce permanent disability. The low incidence of cervical spine injuries has lead to a lack of emergency management experience of on-site medical staff. This paper will review the numerous injuries sustained by the cervical spine in football players and provide insights into prevention and guidelines for return to play. PMID- 11409240 TI - Vertebral osteomyelitis. PMID- 11409241 TI - Gender disparity of anterior cruciate ligament injury. Etiological theories in the female athlete. AB - The gender disparity of anterior cruciate ligament injury has recently been demonstrated and is striking. Numerous etiological factors have been implicated but definitive causation has not yet been determined. An overview of the various hypotheses has been presented and it appears that the etiology is multifactorial. The intrinsic factors are primarily anatomical and cannot be altered in an effort to reduce injury rate. Presently, neuromuscular factors appear to be the most contributory to female ACL injury. Physicians, trainers, and coaches participating in the care of female athletes should encourage proper conditioning. A particular emphasis should be placed on hamstring and gastrocnemius strengthening, as female athletes tend to be quadriceps dominant. Movement training that encourages landing and pivoting with increased knee flexion should also be emphasized. Structured plyometric and jump training programs have been documented to significantly reduce the rate of female ACL injury and continued research in this area may provide more explanation to the gender disparity of ACL injury rates. PMID- 11409242 TI - ACL injuries in the skeletally immature patient. AB - Anterior cruciate ligament injuries are becoming increasingly prevalent in a younger, more athletic population. These injuries require aggressive management given the poor natural history with regard to progressive meniscal damage and advanced degenerative changes. In general, strict nonoperative management has been associated with failure in the individual committed to return to an active lifestyle. As a result a general algorithm can be expressed as follows: 1. Activity limitations and bracing are initially used for the asymptomatic skeletally immature patient with a complete ACL tear. 2. If the patient remains asymptomatic, formal reconstruction can be delayed until skeletal maturity. 3. For the symptomatic patient a precise delineation of their physical development or skeletal age is important. 4. In patients who have reached Tanner IV secondary sexual development or a skeletal age of 13-14 (F/M), a traditional transphyseal reconstruction with hamstring autografts is appropriate. 5. In the skeletally immature patient who has significant skeletal development ahead of them, the judicious use of physeal sparing procedures is an option for the surgeon comfortable with these techniques. In this case the goal is temporization until formal transphyseal reconstruction can be performed after the peak of skeletal growth. PMID- 11409243 TI - Bioresorbable materials in orthopaedics. PMID- 11409244 TI - Another one pays up. HealthSouth settles Medicare fraud charges in whistleblower case. PMID- 11409245 TI - No bargaining. Docs' specialty society, unlike AMA, opposes collective negotiations for residents. PMID- 11409246 TI - CHW's unforeseen challenges. Daughters defection would take profits, but top executive stays bullish. PMID- 11409247 TI - Hearts racing. CIHS announces it will be first to open specialty hospital in Indianapolis area. PMID- 11409248 TI - Hospital exodus. Across the nation, hospital systems dump struggling managed-care plans. PMID- 11409249 TI - Talking points. Some wary of Blues association's guidelines for working with physicians. PMID- 11409250 TI - In the palms of their hands. WebMD's turnaround may depend on docs' acceptance of new technology. PMID- 11409251 TI - Making Medicaid malleable. Governors seek relaxed rules to cover new groups, add some cost-sharing. PMID- 11409252 TI - Channeling calm. Hospitals turn to innovative TV to promote healing in their patients. PMID- 11409253 TI - Minding hospitals' business. Purchasing coalition pushes hospitals to improve patient safety through process measures, but industry says standards are too expensive. PMID- 11409254 TI - Web survey. April survey results: healthcare in short supply. PMID- 11409255 TI - Auditing the auditors. SEC scrutinizes accounting firms for possible client conflicts. PMID- 11409256 TI - Oh no, not that type of review. JCAHO singling out more hospitals for noncompliance with standards. PMID- 11409257 TI - Holding hospitals accountable. National Quality Forum says hospitals should report 'never events' to database. PMID- 11409258 TI - Health plans not reporting adverse actions. PMID- 11409259 TI - Cold-induced urticaria. AB - Cold-induced urticaria is a form of physical urticaria that develops on cold exposure in susceptible individuals. The majority of cases have an unknown etiology (primary or idiopathic). In rare forms, it is associated with various disease entities (secondary). The workup of a patient includes a history and physical examination, cold challenge, and a battery of laboratory tests. The management of such patients includes precaution and avoidance measures as well as treatment with antihistamines. Life-threatening symptoms necessitate carrying a self-administered injectable epinephrine. PMID- 11409260 TI - A retrospective study of risk factors for repeated admissions for asthma in a rural/suburban university hospital. AB - In the study reported, the authors examined risk factors for repeated hospital admissions for asthma in a rural/suburban setting. Charts of patients who were hospitalized two or more times with the diagnosis of asthma between June 1991 and January 1998 were reviewed. A questionnaire was completed for each admission for 65 patients. The results demonstrated an equal male-to-female ratio, with a mean age of 27 years. Hispanics represented 12% of the patients although they accounted for only 2.5% of the general population in the area under study. The mean number of hospital admissions was 3.2. A history of depression existed in 25% of the patients. Noncompliance was admitted in 38%. Twenty-five percent were active tobacco smokers. Acknowledged triggers of asthma included viral infections (74%), exercise (50%), weather conditions (43%), dust (38%), cats (36%), sinusitis (32%), pollen (32%), gastroesophageal reflux disease (31%), dogs (30%), smoke (28%), and emotional stress (15%). Medications at time of admission included albuterol (98%), salmeterol xinafoate (26%), theophylline (38%), ipratropium bromide (55%), nedocromil sodium (20%), cromolyn sodium (35%), prednisone (49%), and inhaled corticosteroids (69%). Ninety-five percent had access to a primary care physician. Fifty-seven percent had a pulmonary and 11% had an allergy consult. These data suggest that patients in rural/suburban areas with repeated hospitalizations for asthma have a high probability of noncompliance, depression, and allergenic triggers. Gastroesophageal reflux was a common recognized trigger. Inhaled steroids were underused, whereas ipratropium and theophylline were overused. Bilingual education on asthma and triggers and social support are necessary even in rural healthcare settings without a large minority population. PMID- 11409261 TI - Allergy test results of a rural and small-city population compared with those of an urban population. AB - The frequency of sensitization to environmental antigens changes in different regions. As such, the pattern of sensitivity to common allergens was studied at multiple sites across central Pennsylvania, an area composed of small cities and rural communities, to determine uniqueness of allergies in populations from this area in contrast to allergies as determined by skin testing in large urban centers. The study reported was undertaken to determine allergen variation from an urban population compared with a rural population of a Northeastern state so that environmental avoidance and immunotherapy can be more precisely prescribed. Patient charts were retrospectively reviewed to determine sensitivity to house dust mites (Dermatophagoides pteronyssinus, Dermatophagoides farinae), cockroach, Penicillium, Aspergillus spp, dog, cat, timothy grass, ragweed, oak, and Alternaria tenuis at five sites in Pennsylvania. All of these sites were classified as "small city" or "rural" for the study. One hundred patient records were examined at each site for the results of allergy testing by the prick puncture, radioallergosorbent test (RAST), or intradermal methods. These small city and rural data were pooled and compared with that of the National Cooperative Inner-City Asthma Study (NCI-CAS), which included 1286 patients from urban environments. The prevalence of allergy to both species of dust mites, dog, timothy grass, and ragweed was significantly greater in the pooled rural group than in the NCICAS inner-city patients (P < .05). In contrast, sensitivity to cockroach antigens and Alternaria was significantly greater in the NCICAS urban population than in the pooled rural group (P < .05). No statistically significant difference was found between the NCICAS and the pooled rural patients in reference to Penicillium, cat, and oak (P > .05). PMID- 11409262 TI - Sinusitis in children: the importance of diagnosis and treatment. AB - The complications from untreated sinusitis in a 10-month-old male infant, though at the more severe end of the spectrum, brings to light the importance of diagnosis and treatment even in the very young patient. Acute sinusitis should be diagnosed using established guidelines. Appropriate pharmacologic and osteopathic manipulative treatment should be initiated on diagnosis. Initial antibiotic therapy is a 14-day course of amoxicillin. If the sinusitis fails to resolve, a trial of a second-line antibiotic should be considered. The use of adjunctive medications such as antihistamines, decongestants, and nasal steroids remains controversial. If the patient fails maximal medical therapy, a computed tomography scan and referral to an otolaryngologist for possible surgical intervention should be considered. PMID- 11409263 TI - Thank God for noncompliance--this time, at least. PMID- 11409264 TI - When it's wise to hire your own attorney. PMID- 11409265 TI - A slip in net worth. PMID- 11409266 TI - Visions of death in the dentist's chair. PMID- 11409267 TI - Washington forecast: much talk. Any action? PMID- 11409268 TI - Let's demand full disclosure by HMOs. PMID- 11409269 TI - Is a "mystery shopper" lurking in your waiting room? PMID- 11409270 TI - Now I understand grief. PMID- 11409271 TI - 5 new-practice pitfalls. PMID- 11409272 TI - For and against privacy regs. PMID- 11409273 TI - LIS shake-out. PMID- 11409274 TI - Incremental approach to electronic medical records. PMID- 11409275 TI - So happy together. PMID- 11409276 TI - Managing change. PMID- 11409277 TI - Analytics and the data warehouse. PMID- 11409278 TI - Identity management--at the heart of CRM (customer relationship management). PMID- 11409279 TI - Good isn't enough. PMID- 11409280 TI - Project management. PMID- 11409281 TI - Inside a virtual nursery. PMID- 11409282 TI - Solving access problems. PMID- 11409283 TI - Small hospital, big Website. PMID- 11409284 TI - Can you keep a secret? PMID- 11409285 TI - What works. Powering the Arizona Telemedicine Program. PMID- 11409286 TI - What works. No waiting. PMID- 11409287 TI - What works. Found money. PMID- 11409288 TI - What works. Working wonders. PMID- 11409289 TI - Laboratory information systems HotList. PMID- 11409290 TI - The case for capitalism in healthcare. PMID- 11409291 TI - HIV surges among young black males. PMID- 11409292 TI - Perspectives. Nursing sentinel events key to quality, safety. PMID- 11409293 TI - Molecular and clinical aspects of mitochondrial diabetes mellitus. AB - This review provides a compact overview on the contribution of mutations in mtDNA to the pathogenesis of diabetes mellitus, with emphasis on the A3243G mutation in the tRNA(Leu, UUR) gene. This mutation associates in most individuals with maternally inherited diabetes and deafness (MIDD) whereas in some other carriers the MELAS syndrome or a progressive kidney failure is seen. Possible pathogenic mechanisms are discussed especially the question why particular mutations in mtDNA associate with distinct clinical entities. Mutations in mtDNA can affect the ATP production, thereby leading to particular clinical phenotypes such as muscle weakness. On the other hand mtDNA mutations may also alter the intracellular concentration of mitochondrial metabolites which can act as signalling molecules, such as Ca or glutamate. This situation may contribute to the development of particular phenotypes that are associated with distinct mtDNA mutations. PMID- 11409294 TI - The metabolic syndrome in twins: a consequence of low birth weight or of being a twin? AB - BACKGROUND: To evaluate if being a twin is a condition at risk of developing metabolic abnormalities in the adult life 'per se' or as a consequence of the lower birth weight. METHODS: 48 monozygotic twins and 50 non-twin controls, both with parental diabetes, were compared; the two groups were comparable for age, sex and body mass index. RESULTS: Twins showed higher values of blood pressure, triglycerides, insulin resistance, fasting insulin, insulin AUCs and higher prevalence of the metabolic syndrome, and, inversely, lower birth weight. After adjustment for birth weight, no significant association was evident with the twin status. Birth weight was inversely associated with the presence of the metabolic syndrome or of at least one or more of its components, while the positive association with the twin status was not significant. CONCLUSIONS: Our data suggest that low birth weight is more relevant than the twin status as a risk factor for the metabolic syndrome and its components. PMID- 11409295 TI - Paraoxonase 1 192 Gln/Arg gene polymorphism and cerebrovascular disease: interaction with type 2 diabetes. AB - Paraoxonase 1 (PON1) is an HDL-associated enzyme which protects HDL and LDL particles from lipid peroxidation. Its enzymatic serum activity varies 10-40-fold between individuals, and its biallelic gene polymorphism at codon 192 (glutamine- >arginine, Gln/Arg) has been associated with coronary artery disease in diabetic patients. To evaluate the role of this PON1 gene polymorphism in cerebrovascular disease, we determined the PON1 192 genotype in 149 patients with hemodynamically relevant extracranial artery stenosis and in 241 controls. The PON1 192 Gln/Arg genotype was determined using polymerase chain reaction followed by Alw I digestion and polyacrylamide gel electrophoresis. Among all subjects, there was no association between the PON1 192 Gln/Arg genotype and cerebrovascular disease (Odds ratio for Arg/Arg and Gln/Arg vs Gln/Gln 0.99, 95%-CI 0.70-1.39). In contrast, in the subgroup of type 2 diabetic patients the PON1 192 Arg allele conferred about twice the risk of cerebrovascular stenosis compared to those homozygous for the Gln allele (Odds ratio 2.00, 95%-CI 0.92-4.38). Our data indicate that in the general population the PON1 192 Gln/Arg gene polymorphism cannot be regarded as a major risk marker for cerebrovascular disease. The observed interaction with type 2 diabetes, however, is supporting the hypothesis that the effect of the PON1 192 Arg allele on atherosclerosis is modulated by other risk factors like diabetes. PMID- 11409296 TI - Prospective study on plasma clotting parameters in diabetic children--no evidence for specific changes in coagulation system. AB - In a prospective study, clotting parameters of 37 children and adolescents with insulin-dependent diabetes mellitus (type 1 diabetes) were compared with those of a healthy control group. In a longitudinal follow-up over two years we found no statistical difference for most of the coagulation parameters investigated, including single factor analysis and coagulation inhibitors. The duration of type 1 diabetes was of no influence on these parameters. The only difference we found between patients and healthy controls was an elevation of PAI-1 in diabetics: median for PAI-1: 2.12 IU/ml in diabetics (range 0.50-8.40 IU/ml) and 0.84 IU/ml in normal controls (range 0.50-1.78 IU/ml). This difference was of statistic significance (p < 0.002) and also found in newly diagnosed patients. During observation time, none of our patients developed thrombosis or signs of vascular disease. In conclusion, we could not confirm the development of a hyper coagulable state in pediatric diabetics, as it is described for adults with type 1 diabetes mellitus. PMID- 11409297 TI - Association between the P12A and c1431t polymorphisms in the peroxisome proliferator activated receptor gamma (PPAR gamma) gene and type 2 diabetes. AB - Variation in the peroxisome proliferator-activated receptor gamma (PPAR gamma) gene may play a role in the development of type 2 diabetes mellitus. Therefore we investigated the association between the P12A and c1431t polymorphisms in the PPAR gamma gene and type 2 diabetes. The incidence of the P12A polymorphism was determined by PCR-RFLP and the c1431t by single-strand conformation polymorphism analysis in 219 patients with, and 429 without type 2 diabetes. The frequency of the A allele of P12A polymorphism was 0.16 and the t allele of c1431t polymorphism, 0.13 in patients with type 2 diabetes, and 0.13 and 0.12 respectively in subjects without diabetes 3.2% of patients with and 1.4% without type 2 diabetes were A12A. Since the polymorphisms are not linked the association of the 9 possible genotypes with type 2 diabetes was determined. All patients with genotype A12A/c1431c had type 2 diabetes (n = 3, p = 0.038). There was no association between A12A/t1431t and diabetes. DNA sequencing revealed no additional mutations in the coding region of the PPAR gamma gene in genotypes A12A/c1431c or A12A/t1431t. The associations found between polymorphisms in the PPAR gamma gene and type 2 diabetes suggest that either the A12 isofrom is functional leading to a predisposition to type 2 diabetes in homozygotes or that there is a third, unknown mutation linked to the A12/c1431 haplotype which is responsible. PMID- 11409298 TI - Stimulatory and synergistic effects of luteinising hormone and insulin like growth factor 1 on the secretion of vascular endothelial growth factor and progesterone of cultured bovine granulosa cells. AB - Vascular endothelial growth factor (VEGF) is the most important factor in the regulation of angiogenesis. Associated with luteinisation and formation of corpus luteum (CL) are alterations in luteal vascularity. The aim of the study was to test under in vitro conditions the stimulation of VEGF and progesterone (P) secretion of bovine granulosa cells by LH, IGF1 (insulin like growth factor) or by factors known to be produced by luteinised granulosa cells or in the early CL. Localisation of VEGF protein in preovulatory follicle and early CL were achieved by immunohistochemistry. LH and IGF1 stimulated dose dependently and significantly P and VEGF when tested alone. Both hormones added simultaneously had clear additive and even more interesting far greater (synergistic) effects on P with LH (0.1 ng/ml) plus 5 or 10 ng IGF1. In contrast, VEGF was stimulated only additively with 0.1 ng/ml of LH plus 5 or 10 ng IGF1. But with the higher dose of LH (1 ng/ml) additionally to the additive effect a tendency for a synergistic action (which was significant with 1 ng LH plus 5 ng IGF1/ml) was observed. Endothelin, oxytocin, progesterone, atrial natiuretic peptide, angiotensin II, prostaglandin F2 alpha alpha, prostaglandin E2, cortisol, fibroblast growth factor 1 and 2 and growth hormone showed no effect neither on P nor on VEGF. Tumour necrosis factor alpha (TNF alpha) stimulated (P < 0.05) VEGF with 10 or 100 ng/ml but not P. TPA (12-0 tetra decaenoyl-phorbol-13-acetate) or Ca2+ ionophore did not show a stimulatory effect in contrast to forskolin which increased P and VEGF secretion dose dependently. The VEGF protein was localised in follicle (granulosa cells, theca cells and some endothelial cells) and early (about 24 h after ovulation) CL (granulosa-lutein cells and endothelial cells). The same signalling pathway by stimulation of cAMP production and proteinkinase A activation for luteinisation and neo-vascularisation demonstrates a close temporal and spatial relationship of these normal physiological processes. PMID- 11409299 TI - Bioavailability of iodine from normal diets rich in dairy products--results of balance studies in women. AB - During the last decade the iodine supply in Germany has increased significantly, but there is still a high frequency of goitre. Therefore the question of iodine bioavailability has arisen. In a two-period study 12 women were given a mixed diet of ordinary foods with milk and milk products of different batches. None of the volunteers suffered from an iodine deficiency according to WHO-criteria. Each period ended with a 9-day balance-study protocol in which all foods were provided. Food and fluid intake were registered, and urine and faeces were quantitatively collected. The iodine content was determined by ICP-MS. The mean intake in the form of solid food amounted to 175 +/- 10 micrograms I/d and to 27 +/- 15 micrograms I/d in fluid form. Milk and dairy products represented the main source of iodine (37%). Iodine was predominantly excreted in the urine (89%, 171 +/- 45 micrograms I/d) and the faeces 11% (20 +/- 11 micrograms I/d). The resulting iodine balance was approximately . In one case an iodine-rich erythrosine preparation with a low iodine bioavailability was used. Between the two periods of consuming different batches of milk and milk products no differences were observed concerning the high bioavailability of iodine. PMID- 11409300 TI - Estrone, but not 17 beta-estradiol, attenuates kainate-induced seizures and toxicity in male mice. AB - Estrogens change the susceptibility to seizures in humans and experimental animals. In this study, the effect of estrone and 17 beta-estradiol on kainate induced seizures and neurotoxicity was investigated in male mice. Pre-treatment with estrone (250-1000 micrograms/kg) at 24 and 2 hours before kainate (40 mg/kg) administration significantly decreased both the percentage of animals with clonic seizures and their mortality (the latter at a dose of 1000 micrograms/kg only). On the other hand, 17 beta-estradiol (10-500 micrograms/kg) had no effect on seizures, and its dose of 10 micrograms/kg increased mortality. When given alone at a dose of 1 mg/kg, tamoxifen, an antagonist at estrogene receptors, did not affect the kainate-induced seizures, but prevented the anticonvulsant effect of estrone. A histological analysis showed that 73% of mice injected with vehiculum and kainate incurred hippocampal damage. Estrone (2000 micrograms/kg) decreased the percentage of animals with hippocampal neuronal loss down to 43%, and that effect was not antagonized by tamoxifen. Pretreatment of mice with 17 beta estradiol had no effect on the kainate-induced neuronal loss. Additionally, we found that kainate injected i.p. had a profound effect on the immune system of mice, as reflected by a decrease in the thymus weight and an increased metabolic activity of splenocytes. The anticonvulsive dose of estrone (1000 micrograms/kg) did not change the immunoreactivity of either control or kainate-treated mice. In conclusion, the obtained data indicate that estrone, but not 17 beta-estradiol, attenuates the kainate-induced seizures, mortality and excitotoxicity in male mice. Moreover, it is suggested that the suppressive effect of estrone on clonic seizures involves intracellular receptors, whereas its antineurotoxic activity seems to depend on a non-genomic mechanism. PMID- 11409301 TI - Short-term modulation of prolactin secretion by melatonin in anestrous ewes following dopamine- and opiate receptor blockade. AB - This study tested the hypothesis that the dopaminergic and opioidergic systems are not involved in the short-term stimulatory action of melatonin (MLT) on the secretion of prolactin in anestrous ewes. Thus, MLT should stimulate prolactin release after blockade of either dopamine (DA) or opiate receptors with specific antagonists at the level of the pituitary gland and central nervous system (CNS), respectively. During afternoon intracerebroventricular (icv.) infusion of MLT, the mean plasma prolactin concentration increased significantly (P < 0.001) as compared with the concentrations noted before and during the infusion of the vehicle (veh.). As a result of subcutaneous (s.c.) injection of sulpiride (SULP, DA antagonist), an increase in plasma prolactin concentration was observed, followed by a gradual decrease during the icv. infusion of the vehicle. MLT infused icv. significantly increased (P < 0.001) the secretion of prolactin in SULP + MLT-treated ewes, as compared with the concentration of prolactin noted during infusion of the vehicle in SULP + veh.-treated ewes. Naloxone (NAL, opioid antagonist) infused icv. did not significantly affect the secretion of prolactin, however, a significant (P < 0.01) increase in the concentration was observed after the infusion. In MLT + NAL-treated ewes, the plasma prolactin concentration increased significantly (P < 0.001) during the infusion, as compared with the concentration noted before and that in NAL-alone infused ewes. These results demonstrate that melatonin stimulates prolactin release after the pharmacological exclusion of the dopaminergic input with the DA antagonist sulpiride and also despite the presence of DA activity in the hypothalamus after NAL treatment. Secondly, endogenous opioid peptides are not a major component of this melatonin action. PMID- 11409302 TI - Further evidence for direct vascular actions of statins: effect on endothelial nitric oxide synthase and adhesion molecules. AB - The effect of the synthetic statin fluvastatin was investigated on the concentrations of endothelial nitric oxide synthase (eNOS) and of soluble adhesion molecules in human vascular endothelial cell cultures. Fluvastatin was able to increase dose-dependently eNOS concentration. In addition, fluvastatin reduced concentrations of E-Selectin and ICAM-1. These results further support the evidence that statins elicit important effects on the vascular endothelium which may contribute to the reduction of morbidity and mortality of cardiovascular diseases. PMID- 11409303 TI - Asymptomatic hypercalcemia due to an ectopic parathyroid adenoma in an 8-year-old boy. AB - The diagnosis of primary hyperparathyroidism in children is often delayed and is usually based on symptoms of hypercalcemia rather than abnormal laboratory values alone. We report the case of an 8-year-old boy with hypercalcemia, hypophosphatemia and mildly, but inadequately elevated intact parathyroid hormone (iPTH) who presented without any symptoms of hyperparathyroidism. Although imaging studies were misleading and four normal parathyroid glands were found intraoperatively, exploration of the thymus revealed an ectopic parathyroid adenoma. After removal of the ectopic gland, a rapid iPTH immunoassay proved immediate normalization of iPTH. This is the first report of sporadic isolated primary hyperparathyroidism diagnosed in an asymptomatic child on the basis of hypercalcemia and hypophosphatemia. PMID- 11409304 TI - Endoscopic endonasal pituitary surgery: evolution of surgical technique and equipment in 150 operations. AB - OBJECTIVE: The evolution of the senior author's (HD Jho) surgical experience on endoscopic endonasal transsphenoidal pituitary surgery is reviewed in order to introduce a standardized surgical technique for pituitary endoscopy. METHODS: The progressive evolution of the surgical technique and the development of surgical instrumentation in transsphenoidal endoscopy is reviewed based upon the experience of more than 150 operations performed by the senior author between the years 1993 and 1998. RESULTS: An endoscope was used to assist visualization during conventional microscopic surgery in the first four cases (endoscope assisted microsurgery). Subsequently, endonasal pituitary endoscopy was performed via a nostril. When the endonasal route was adopted, it eliminated the use of a transsphenoidal retractor. The use of vasoconstrictors and any form of nasal packing was discovered to be unnecessary. The inferior margin of the middle turbinate was such a consistent surgical landmark leading to the sella that the use of an intraoperative fluoroscopic C-arm was also eliminated. The adoption of a septal breaker, variously angled suction cannulas and suction-coagulators has made the operation cleaner, easier and faster. As experience increased, the operation time progressively shortened and mucosal trauma became minimal. The median patient hospital stay was one night and postoperative discomfort was noted to be minimal. CONCLUSIONS: The evolution of the senior author's endoscopic pituitary surgery is reported with a description of our current standardized surgical technique. PMID- 11409305 TI - Minimally invasive craniotomy using the Steiner-Lindquist stereotaxic guide. AB - Thirty-three obscure intracranial lesions were located using the Steiner Lindquist microsurgical stereotaxic guide and then surgically resected. Seventeen of the lesions were located in the parietal region, six in the frontal region, three in the parietooccipital region, three in the temporoparietal region, one in the thalamic region, one in the centrum semiovale, one in the brainstem, and one in the third ventricle. Twenty-three lesions were in subcortical or cortical locations. In 28 cases, the lesion was totally removed, while in 5 the lesion was subtotally resected. Pathological examinations confirmed glial tumor in eight patients, metastasis in seven, meningioma in two, cavernous angioma in eight, arteriovenous malformation (AVM) in four, hematoma in two, dysembryoblastic neuroepithelial tumor in one, and septum pellucidum cyst in one. Two patients developed transient complications postsurgery. Mean lesion size was 23 +/- 0.97 mm. The hospitalization period ranged from 1 to 6 days (mean 3.4 +/- 1.3 days). Surgeries were performed under general anesthesia, or under local anesthesia with the patient awake. The Steiner-Lindquist microsurgical stereotaxic guide is useful for pinpointing small lesions, especially those in the subcortical and deep areas. Knowing the precise location of the lesion facilitates removal through a small craniotomy incision. This minimally invasive procedure reduces the number of postoperative neurological complications, and also cuts costs by shortening the hospital stay. PMID- 11409306 TI - Eyebrow incision using tattoo for anterior fossa lesions: technical case reports. AB - Some Korean women draw tattoo in their eyebrow to cover the scant hair. If a patient has a tattoo in her eyebrow, lesions of anterior cranial fossa can be easily managed with this small eyebrow skin incision through a small unilateral supraorbital craniotomy. By this technique, 7 cases of anterior cranial fossa lesions were successfully treated without any major complications. This leads to less facial scar, less operation time and an earlier return to social adaptation. PMID- 11409307 TI - Endoscopic surgery for large posterior fossa arachnoid cysts. AB - The authors report two cases of large arachnoid cysts of the posterior fossa treated by endoscopic surgery. One patient underwent a successful endoscopic cyst fenestration by burr hole approach after several procedures of shunt revision. In another an endoscope-assisted microsurgical intervention was necessary. Lateral (cerebellar or cerebellopontine angle) cysts, as two reported cases, may be treated through a lateral retromastoid approach by fenestration into the prepontine cistern and eventually into the cisterna magna. We advise to start the operation through a burr hole and to try to realize the fenestration by endoscopy only. If this attempt fails, an endoscope-assisted microsurgical technique may be performed by enlarging the craniectomy. In this last instance the endoscope is useful particularly deeply to fenestrate the anterior cyst wall in the prepontine or ambient cisterns, where it provides more illumination and helps to identify the nervous and vascular structures. PMID- 11409308 TI - Endoscopic treatment of hydrocephalus in children: a controlled study using newly developed Yamadori-type ventriculoscopes. AB - Although cerebrospinal fluid (CSF) shunting is the most common neurosurgical treatment for hydrocephalus, the long-term results have still been unsatisfactory because of a wide variety of shunt complications. We have recently developed flexible ventriculoscopes (Yamadori-type) which have excellent image quality, maneuverability, and capabilities for endoscopic operation. Here we report the efficacy of the new treatment in 88 children with hydrocephalus who initially underwent either ventriculoscopic operation or shunting surgery. The primary outcome measures were the rate of shunt independency and/or shunt complications with a follow-up of 2 years in each group. We performed endoscopic third ventriculostomy in cases of aqueductal stenosis, cyst fenestration, and choroid plexus coagulation in limited cases of communicating hydrocephalus. Overall, thirty-three (75%) of the 44 children initially treated endoscopically did not require ventriculoperitoneal (VP) shunts. The endoscopic procedures were repeated in the remaining 11 children (25%) mostly less than 1-year-old who ultimately required endoscope-guided VP shunting. Even in such patients, there was virtually no need for shunt revisions and no major complications such as slit-like ventricle, meningitis, and intraventricular hemorrhage. These results were statistically highly significant (p < 0.0001) compared to a control group of 44 patients treated initially by VP shunting. Our data demonstrate that therapeutic ventriculoscopy is safe and clinically effective as the first-line treatment of hydrocephalus in children. PMID- 11409309 TI - Association between decreased disc signal intensity in preoperative T2-weighted MRI and a 5-year outcome after lumbar minimally invasive discectomy. AB - A total of 39 patients who had undergone microdiscectomy or percutaneous nucleotomy for lumbar disc herniation were examined after a follow-up of 5 years. The overall outcome was satisfactory in 80% of the patients treated, and only 1 (3%) patient had been reoperated during the follow-up. Clinical signs and symptoms of lumbar instability were detected in 10 (26%) patients. All these 39 patients had been examined with lumbar magnetic resonance imaging (MRI) on the day preceding the operation; the presence of disc degeneration was graded as severe, mild or non-existent depending on the visual brightness of the discs on T2-weighted images, as compared to the signal intensity of the lumbar vertebrae. None of the 12 patients with no preoperative disc degeneration in MRI suffered from postoperative clinical signs and symptoms of instability as compared to 10 (37%) of the 27 patients with mild or severe disc degeneration suffering from instability (p = 0.04). Thus, the results of the present study imply that the grade of the disc degeneration in preoperative T2-weighted MR images significantly predicted the occurrence of postoperative clinical instability. PMID- 11409310 TI - Stereotactic guided laser-induced interstitial thermotherapy (SLITT) in gliomas with intraoperative morphologic monitoring in an open MR-unit. AB - Stereotactic guided laser-induced interstitial thermotherapy (SLITT) represents a minimal invasive method to produce necrosis in cerebral tumor tissue by local heating. The dose/response relationship relies on experimental studies and few clinical data performed in high field MR systems. A better understanding of the energy-dose/tissue response in human brain tumors is important to optimize this treatment modality. Twenty-four patients with gliomas were treated with SLITT, with a total of 30 laser procedures performed. Under local anesthesia 600 microns laser-fibers were inserted by stereotactic-guided technique into the center of the tumor. In a low field open MR system (0.2 T) the denaturation of the tumor using a neodymium YAG laser (1064 nm) was monitored by 3D-turbo FLASH T1-weighted sequences. Laser energy was applied in steps of 400 to 1200 Joules. Development of necrosis at a mean total energy dose of 2979 Joules could be monitored in all procedures. Two different thermal lesion architectures were observed. First signal changes were monitored after a mean of 1108 Joules and 1393 Joules, respectively. Mean max. total lesion size was 21.2 mm. The higher the total energy the larger was the thermolesion, but no linear relationship could be seen. Tumor tissue response showed no dependency on tumor grading. Monitoring of stereotactic guided laser-induced thermolesions in the low-power MR OPEN is feasible and safe. Although lesion size basically is energy dependent, it should be applied individually, since the thermal response in brain tumors varies due to different optical properties, even in the same tumor gradings. PMID- 11409311 TI - Successful radiosurgical treatment of lesional epilepsy of mesial temporal origin. AB - We report a case of successful radiosurgical treatment of lesional epilepsy of mesial temporal origin. A patient presented with a 2-year history of medically intractable complex partial seizures associated with a mesial temporal angioma. Interictal scalp EEG and MEG showed focal epileptiform activity around the lesion. 99mTc-HMPAO-SPECT and 18F-FDG-PET demonstrated depressed blood flow and glucose metabolism in the corresponding temporal lobe. The patient underwent gamma knife radiosurgery for the causative lesion with a low marginal dose of 18 Gy. After treatment, the partial attack ceased without shrinkage of the lesion or peri-lesional parenchymal radiation injury. PMID- 11409312 TI - A pair of clamps for a safe removing and repositioning of beta and gamma angular settings of Brown-Roberts-Wells stereotactic system during operation. AB - The Brown-Roberts-Wells arc system is a non-target-centered design, i.e., without an independent approach angle. The approach angle of this system strictly depends on precalculated values (entry and target point). Therefore, some components of the system used sometimes prevent a direct insight into the operation field. Once the entry point has been set, the arc system normally has to be taken off to permit an unimpeded approach to the burr hole. To facilitate rotation and return to the primary beta and gamma angular settings during stereotactic craniotomy and other surgery, a pair of clamps was designed for the BRW arc system. These clamps help the approach to the entry point in such a way that some components of the arc (e.g., the guide block holder) are removed from the surgical field, thus giving wide visual access for the stereotactic approach. Consequently, it is no longer necessary to remove the entire arc system, resulting in an increased operation safety and shorter operation times. PMID- 11409313 TI - Effects of electrical stimulation of the Gasserian ganglion on regional cerebral blood flow after induced subarachnoid hemorrhage in pigs evaluated by 99mTc-HMPAO SPECT. AB - It could be demonstrated that cervical spinal cord stimulation increases cerebral blood flow. The effects of electrical stimulation of the trigeminal ganglion in the acute phase of SAH in pigs were investigated. The experiments were carried out on 11 domestic pigs divided in two groups (group I: SAH [n = 5]; group II: SAH and trigeminal stimulation [n = 6]). In all animals a native SPECT was performed. The Gasserian ganglion was exposed for inserting the stimulation electrode. SAH was induced by injecting 10 ml autologous blood through a catheter placed in the suprasellar cistern. 30 minutes after SAH-induction electrical stimulation was started for two hours in group II (2.8-4.5 V, 50 Hz, 300 microseconds). 99mTc-HMPAO (400-540 MBq) was injected intravenously 110 minutes later. In group I 99mTc-HMPAO was applied after the same time interval. 80 minutes later SPECT was performed. Data were processed to calculate the uptake of radioactivity (%/kg tissue weight). The mean values were calculated for the different groups: native animal examination (%/kg tissue weight): 0.6343; group I: 0.468; group II: 0.6533. Comparing the mean values a highly significant difference between group I and group II (p < 0.01) and between native examination and group I (p < 0.01) could be found. No statistical significance could be detected on comparing the left/right-ratio in any ROI. The electrical stimulation of the Gasserian ganglion leads to a significantly increased uptake of 99mTc HMPAO after induced SAH. Maybe the stimulation of the Gasserian ganglion constitutes a new therapeutic modality treating disturbed rCBF after SAH. PMID- 11409314 TI - Subependymal giant-cell astrocytoma in tuberous sclerosis: endoscopic images and the implications for therapy. AB - Supratentorial intraventricular tumors are not frequently encountered in childhood. One of the most frequent intraventricular glial tumors is the subependymal giant-cell astrocytoma, mostly associated with tuberous sclerosis. These tumors are diagnosed on computed tomography (CT) or magnetic resonance imaging (MRI) scans. They can occur isolated or multiple and operative resection is advised if these tumors cause symptoms, usually raised intracranial pressure due to obstructive hydrocephalus. However, the number of tumors can be much higher than seen on radiological examination making total resection of all tumors impossible. We demonstrate this with the endoscopic images derived during the endoscopic removal of a subependymal giant-cell astrocytoma obstructing a foramen of Monro in a 15-year-old boy with tuberous sclerosis. PMID- 11409315 TI - High incidence of chromosome aberrations after radiochemotherapy for Hodgkin's disease: a report of a case and a review of the literature. AB - To emphasize the importance of information obtained by conventional cytogenetic tests, in this study we will present a case report showing the development of HD in a young engineer of medical radiology, who had been professionally exposed to ionizing radiation during 4 years. Bio-dosimeter data and chromosomal aberration analysis made in a group of colleagues working in the same ionizing conditions excluded the possibility that she was overexposed to ionizing radiation, but retroactive analysis showed that at the time of employment she had a moderately increased level of chromosomal aberrations in peripheral blood lymphocytes (4.6%), with higher than normal incidence of chromatid breaks (22 x 10(-3) per cell) and dicentric fragments (5 x 10(-3) per cell). After the treatment of lymphoma by chemotherapy in combination with radiotherapy she also demonstrated a very high post-therapeutic level of chromosomal aberrations (21.5%), which consisted mostly of increased dicentric fragments (65 x 10(-3) per cell). Although her illness is now clinically cured, the observed genotoxic changes point to a greater risk for delayed complications after HD, emphasizing the necessity for further continuous survey of this patient. PMID- 11409316 TI - Monoclonal antibodies specific to polynuclear aromatic hydrocarbons. PMID- 11409317 TI - Monoclonal antibodies specific for bovine CD18. PMID- 11409318 TI - Patterns of deletions and the distribution of breakpoints in the dystrophin gene in Czech patients with Duchenne and Becker muscular dystrophy (statistical comparison with results from several other countries). AB - Deletion pattern analysis of the dystrophin gene was performed in 115 unrelated Czech patients with Duchenne and Becker muscular dystrophy. In 50 patients (43.5% of the analysed patients) exon deletions were detected by routinely performed multiplex PCR for 18 selected exons and for the area of musclespecific promoter of the dystrophin gene. All startpoints and endpoints of deletions (100 breakpoints) were detected using PCRs for another 29 exon areas of the dystrophin gene (altogether primers for 47 different exons were used). Most of the breakpoints were found in introns 44 (16% of breakpoints), 47 (14%) and 50 (8%). The comparison of distributions of breakpoints in the area of the main hot spot of the dystrophin gene (introns 43-52) was made (chi 2 test in a contingency table) in six different populations from the Czech Republic, Bulgaria, Hungary, Italy, Turkey and India. In compared populations, statistically significant differences were found by the pooled test. No significant difference between the Czech population and other studied populations was found by pair comparisons. On the other hand, pair comparisons revealed significant differences between populations from Bulgaria and Hungary, Bulgaria and Turkey, Hungary and Italy. The results of the presented study support the theory suggested by other authors that specific differences in intron sequences of the dystrophin gene can exist between different populations, possibly as a result of a genetic drift. PMID- 11409319 TI - Supernumerary human preembryos provide potential for preimplantation genetic diagnosis. AB - Preimplantation genetic diagnosis is an alternative to the classical prenatal diagnosis for couples undergoing in vitro fertilization. It allows very early embryo selection--before the intrauterine embryo transfer. Prior to clinical application of preimplantation diagnosis in the Infertility Treatment Centre "Technobioassistance", Sofia Medical Faculty, we have developed preimplantation diagnosis models of human spermatozoa and untransferred 2-8-cell human embryos obtained in vitro. Directly fluorescein isothiocyanate-labelled probes specific for the centromeric regions of chromosomes 1, 5, 19 and X (Boehringer Mannheim) were used. Eighty-six point three percent of fixed blastomeres with normal size and shape had unfragmented nuclei with dispersed interphase chromatin or mitotic chromosomes and all of them demonstrated successful hybridization. In cases with more than 75% of embryo cells suitable for analysis we were able to estimate the presence of mosaicism among the blastomeres. PMID- 11409320 TI - Effects of combined treatment of rats with cadmium and ionizing radiation on nucleic acids in the kidneys, liver and haemopoietic organs. AB - The influence of Cd (1 mg/rat CdCl2 i.p.) and/or gamma radiation (6 Gy) on RNA and DNA content and/or concentration in the intact kidney and hypertrophic kidney (on the 44th hour after UN) and in other slowly and quickly proliferating organs was studied. The period between administration of Cd and Ir in the group with combined treatment was 30 min; between treatment (administration of Cd, Ir and combination of both treatments--Cd + Ir) and UN it was 1, 7, 14 and 21 days. The total extent of damage caused by the treatments in the investigated organs was following: intact kidney < liver < hypertrophic kidney and bone marrow < spleen < thymus. In the intact and hypertrophic kidney and liver, the administration of Cd caused more extensive changes in comparison with gamma irradiation, and the effects of combination of the treatments were similar to those of Cd alone. In the bone marrow, spleen and thymus, more profound changes were observed after Ir in comparison with Cd administration, and the effects of combined treatment were similar to the effects of Ir alone. The changes in the hypertrophic kidney after administration of Cd and/or Ir were more extensive than in the intact kidney, which suggests latent injury induction in the rat kidney by these noxa. The higher effectiveness of the treatments in the hypertrophic kidney than in the intact one was manifested mostly by the decrease in the RNA and DNA content, which was mainly due to inhibition of growth induced by UN and not by a real decrease in DNA and RNA contents caused by loss of damaged cells. PMID- 11409321 TI - Differences of ropivacaine and bupivacaine relevant to antiinflammatory activity, platelet aggregation and antioxidant activity in vitro. AB - Ropivacaine and bupivacaine affect the in vitro growth of rat fibroblasts and monkey kidney Vero cells with bupivacaine generally showing the stronger effect. Up to 3 mM concentration the two anesthetics affect the expression of genes differently for CD2, CD3 gamma, CD40L, IL-2, IL-2R alpha, IL-2R beta, IL-2R gamma, IL-4, and IL-4R during activation of human lymphocytes, with bupivacaine showing the higher effect. Human platelet aggregation is inhibited by the two anesthetics which also show an antioxidant effect on lipid peroxidation of rat liver microsomes. In both cases bupivacaine seems more active than ropivacaine. PMID- 11409322 TI - Synthesis, in vitro antiproliferative activity and DNA-interaction of benzimidazoquinazoline derivatives as potential anti-tumor agents. AB - The synthesis of benzimidazoquinazoline derivatives bearing different alkylamino side chains is reported. All new compounds tested by means of an in vitro assay exhibit antiproliferative activity toward human tumor cell lines. The cytotoxic effect depends on the type of side chain inserted in the planar nucleus and in some cases it is comparable to that of the well-known drug ellipticine. In order to understand the mechanism of action of these compounds, the interaction with DNA has been investigated. Linear flow dichroism measurements allowed us to verify the formation of a molecular complex with DNA and the corresponding geometry of interaction. Intrinsic binding constants have also been evaluated by performing fluorimetric titrations. PMID- 11409323 TI - 2-(4-R-phenoxy/phenylthio)alkanoic esters of l-lupinine. AB - Considering the great pharmacological interest in phenoxy/phenylthioalkanoic esters of open-chain or cyclic aminoalcohols, a set of ten such esters of lupinine was prepared. Initially, their ability to displace [3H]QNB from rat brain preparation was investigated. With the exception of two, all the prepared esters exhibited good affinity to muscarinic receptors (on a non-selective basis), with pKi in the range 6.67-7.68. PMID- 11409324 TI - Glutathione uptake after intraperitoneal administration and glutathione radiopharmacology after rectal administration, in mice. AB - Glutathione is a biologic aminothiol radioprotector. Hydrolysis of exogenous glutathione takes place in the extracellular compartment and leads to two metabolites: gamma-glutamylcysteine and glycine. In healthy mice, after an intraperitoneal administration of glutathione, all organs absorb the gamma glutamylcysteine and the glycine with variable kinetics according to their enzymatic equipment. The rectal administration of glutathione in mice previously irradiated at the pelvic region, increases the availability of glutathione in the rectum and in other organs at a distant from the irradiation site. This contribution could be used to protect the rectum and the uterus during therapeutic irradiation. PMID- 11409325 TI - Synthesis and pharmacological evaluation of potent and enantioselective sigma 1, and sigma 2 ligands. AB - In a previous study we found that substitutions of the (+)-cis-N-normetazocine nucleus of (+)-MPCB with 1-adamantanamine provide the compound (+/-)-10 with high affinity and selectivity for sigma receptors. Starting with this result we have synthesized a new series of eight 1-phenyl-2-cyclopropylmethylamines structurally related to (+/-)-10, and binding affinities, with respect to sigma 1, sigma 2, opioid and dopaminergic D2 receptors, have been reported. All compounds showed a negligible opioid and dopaminergic affinity and high selectivity for sigma receptors. Modifications on the amino moiety and methylcarboxyester group of 10 provide compounds with different sigma 1 and sigma 2 binding affinity and selectivity. Moreover, we have also synthesized the respective enantiomers of componds (+/-)-10 and (+/-)-18 in order to evaluate the enantioselectivity for sigma 1 and sigma 2 receptors. The binding data showed that carboxymethylester on the cyclopropane ring was more critical for enantioselectivity than the hydroxymethylenic group. In fact, the (-)-10 enantiomer showed a preference for sigma 1 whereas (+)-10 showed a preference for sigma 2. PMID- 11409326 TI - Affinity labels as tools for the identification of opioid receptor recognition sites. AB - Affinity labels have proven to be useful tools in opioid research. We review experiments carried out with the mu opioid receptor affinity label, beta funaltrexamine (2), that support the concept of different recognition sites for mu opioid agonists and antagonists. The data are interpreted in the context of a dimeric receptor that contains two allosterically coupled binding sites: one that binds endogenous agonist, and the second that functions as an inhibitory modulator of agonism. It is proposed that exogenous antagonists bind selectively to the second site. The first of a new class of affinity labels, PGNA (5), that contains the phthaldehyde moiety attached to an opioid antagonist pharmacophore, is described. This class of ligands has been named 'reporter affinity labels' because covalent association leads to the formation of a fluorescent isoindole that is diagnostic for cross-linking of lysine and cysteine residues. PGNA binds opioid receptors covalently, as suggested by (a) irreversible binding to cloned opioid receptors, (b) irreversible opioid antagonism in the guinea pig ileum preparation, and (c) ultra-long opioid antagonism in mice. Since flow cytometry experiments revealed specific enhancement of fluorescence in cloned mu receptors after a 1 min exposure to 5, it is concluded that covalent binding has occurred via the formation of an isoindole, presumably by cross-linking neighboring lysine and cysteine residues in the vicinity of the receptor recognition site. PMID- 11409327 TI - Studies on indomethacin inserts prepared by water-soluble polymers. II. The relation between dissolution rate and swelling behaviour. AB - Inserts containing indomethacin were prepared using water-soluble polymers such as hydroxypropyl cellulose, methylcellulose, hydroxypropyl methylcellulose and polyvinyl alcohol by the film casting method. According to the different characteristics of the polymer used, these inserts exhibit different release kinetics and swelling behaviour. In this study, we examined the relation between swelling behaviour of the polymer and the release of the indomethacin from inserts. Thus an electrical device for measuring the thickness of the hydrated inserts was developed. The thickness of the hydrated inserts was measured by this electrical device at selected time intervals for release studies. The results were interpreted from normalised increases in thickness of the hydrated insert. The mechanism of drug release was identified by means of the value of the ratio R/F, calculated according to the equation developed by Peppas. When the ratio R/F of the insert decreased, drug release from the insert became diffusive. As the normalised thickness of the insert increased, the rate of drug release decreased. PMID- 11409328 TI - 5-acyloxy-5-hydroxymethyltetrahydro-2-furancarboxylate as a novel template for protein kinase C (PKC) binding. AB - A series of alkyl tetrahydrofuran-2-carboxylates (1-4) bearing a new set of three pharmacophoric groups were tested as protein kinase C (PKC) ligands. The compounds were synthesized from commercially available glycidyl 4-methoxyphenyl ether. The correlation between their binding affinities for PKC-alpha and a conformational fit to phorbol ester indicates they mimic a pharmacophore model comprising the C20-OH, C3-C=O and C9-OH rather than that including the C13-C=O moiety. PMID- 11409329 TI - Pyrocatechol violet in pharmaceutical analysis. Part I. A spectrophotometric method for the determination of some beta-lactam antibiotics in pure and in pharmaceutical dosage forms. AB - A fairly sensitive, simple and rapid spectrophotometric method for the determination of some beta-lactam antibiotics, namely ampicillin (Amp), amoxycillin (Amox), 6-aminopenicillanic acid (6APA), cloxacillin (Clox), dicloxacillin (Diclox) and flucloxacillin sodium (Fluclox) in bulk samples and in pharmaceutical dosage forms is described. The proposed method involves the use of pyrocatechol violet as a chromogenic reagent. These drugs produce a reddish brown coloured ion pair with absorption maximum at 604, 641, 645, 604, 649 and 641 nm for Amp, Amox, 6APA, Clox, Diclox and Flucolx, respectively. The colours produced obey Beer's law and are suitable for the quantitative determination of the named compounds. The optimization of different experimental conditions is described. The molar ratio of the ion pairs was established and a proposal for the reaction pathway is given. The procedure described was applied successfully to determine the examined drugs in dosage forms and the results obtained were comparable to those obtained with the official methods. PMID- 11409330 TI - Chromatographic (GC-MS, HPLC) and virological evaluations of Salvia sclarea infected by BBWV-I. AB - Salvia sclarea cultivated at the Herb Garden of Casola-Valsenio (Emilia-Romagna region, Italy) has been found for the first time naturally infected by broad bean wilt fabavirus, serotype I (BBWV-I). Symptomatic plants showed malformed leaves, with chlorotic mosaic followed by yellowing and stunting. BBWV-I was identified by applying virological tests: mechanical inoculations on herbaceous plants, electron microscopy, DAS-ELISA and PAS-ELISA. The essential oil obtained from BBWV-infected material corresponded to 2/3 the quantity of that from healthy material. The GC-MS and HPLC analyses of these oils afforded a comparative analytical profile of the two plant materials attributed to BBWV-I infection. The oils from infected materials showed higher percentages of sesquiterpene hydrocarbons (e.g. germacrene D and beta-caryophyllene), monoterpene alcohols (e.g. alpha-terpineol) and diterpenoids (mainly sclareol). In contrast, lower levels of monoterpene hydrocarbons (e.g. myrcene, limonene and the two ocimene isomers) and the principal components (linalyl acetate and linalool) were observed. PMID- 11409331 TI - Synthesis, characterization and pharmacological activities of 5,6,11,12 tetrahydroindolo[2,3-alpha]carbazole derivatives. AB - A series of new 5,6,11,12-tetrahydroindolo[2,3-alpha]carbazole derivatives (3a-h) was synthesized. Treatment of 8-methyl-1-oxo-1,2,3,4-tetrahydrocarbazole (1a) with phenylhydrazine hydrochloride in ethanol furnished the title compound (3a) in poor yield along with 8-methyl-1-phenylhydrazono-1,2,3,4-tetrahydrocarbazole (2a). Better yields were obtained when 1a-h were treated with phenylhydrazine in glacial acetic acid. All the newly synthesized compounds were characterized on the basis of IR, NMR, mass-spectra and elemental analysis and screened for pharmacological activities. PMID- 11409332 TI - Synthesis and antinociceptive activity of 6-substituted-3-pyridazinone derivatives. AB - A series of 3-pyridazinones carrying morpholino, arylpiperidino and arylpiperazino moiety in the position 6 IIa-g were synthesized and evaluated for antinociceptive activity. In the modified Koster test in mice 4-(4-fluorophenyl) piperazine, IIf, was found the most active compound. All the compounds except IId were more active than aspirin in the antinociceptive activity test. PMID- 11409333 TI - Carboxylic acids and skeletal muscle chloride channel conductance: effects on the biological activity induced by the introduction of methyl groups on the aromatic ring of chiral alpha-(4-chloro-phenoxy)alkanoic acids. AB - One or two methyl groups have been introduced on the aromatic ring of two chiral clofibric acid analogs, 2-(4-chloro-phenoxy)propanoic and 2-(4-chloro phenoxy)butanoic acids. The biological activity of the derivatives obtained (3-6) has been evaluated on the skeletal muscle chloride conductance (gCl). The results confirm the hypothesis of two different sites modulating chloride channel function, an excitatory site that increases channel activity and an inhibitory site that produces a channel block. In fact, this chemical modification strongly reduces the blocking activity of the (R)- and (S)-enantiomers in comparison with the parent compounds, but does not markedly affect the ability of the (R) enantiomers to increase chloride channel conductance. PMID- 11409334 TI - Doctors become palm readers. PMID- 11409335 TI - I.T. paints a different picture. PMID- 11409336 TI - Many happy returns: measuring ROI. PMID- 11409337 TI - Web technology makes its mark on software. PMID- 11409338 TI - Aging bean counting systems run out of gas. PMID- 11409339 TI - Putting an end to 'inside jobs'. PMID- 11409340 TI - Readers' perspectives. Is the role of consultants increasing or decreasing in importance in health care information technology? PMID- 11409341 TI - Cruel lessons from an epidemic. PMID- 11409342 TI - Morality goes to school. PMID- 11409343 TI - A little bit of sugar.... a major challenge to the potency of placebos. PMID- 11409344 TI - The power of gender. Boys will be boys. PMID- 11409345 TI - Not acting their age. PMID- 11409346 TI - Back to work, on a mission. Retirees are tackling social problems. PMID- 11409347 TI - When drug costs sting, look for relief. PMID- 11409348 TI - Shaping up under a watchful eye. Hospital-based gyms offer medical expertise. PMID- 11409349 TI - [Glutamate receptors and transporters in the brain and peripheral tissues]. AB - Functional diversity of glutamate receptors in the central nervous system (CNS) is a consequence of their considerable molecular diversity. The family of glutamate receptors including their subunits consists of more than 25 proteins. It is the result of gene expression as well as extensive post-transcriptional modifications. Evidence is increasing that glutamate receptors are localised not only in the CNS, but also in the peripheral tissues. Whereas characteristics and physiological significance of peripheral glutamate receptors are little understood, there are studies indicating their role in hormone secretion, neuromuscular functions, sensory transmission and paracrine signalization. In addition, peripheral glutamate receptors may participate in mediating of excitatory amino acids toxicity. A special attention is paid to glutamate receptors localised in the heart and adrenals, as these receptors may be involved in the maintenance of homeostatic mechanisms under pathological or stress conditions. PMID- 11409350 TI - [Effect of nutrition on adipose tissue metabolism in humans]. AB - Lipolysis in adipose tissue and balance between energy intake and expenditure are involved in the regulation of adipose tissue mass. Several recent findings suggest that alterations in the regulation of lipolysis and/or energy balance might contribute to the development of obesity. Hormone-sensitive lipase and uncoupling proteins play important role in regulation of lipolysis in adipose tissue as well as in the regulation of energy balance of various tissues. Mechanisms of the control of expression of genes coding synthesis of these proteins are poorly known. A brief overview of the present knowledge of the effects of nutritional intervention on the regulation of lipolysis in adipose tissue and on the expression of genes of hormone-sensitive lipase and that of uncoupling proteins is given in this article. Results of the authors' studies on the effect of calorie restriction on gene expression in adipose tissue are presented. PMID- 11409351 TI - [Guanylins--a potential new approach in research on postprandial natriuresis]. AB - More than 20% of adults in industrialized countries display arterial pressure outside the normal physiological range. For most individuals, the molecular basis of hypertension remains unknown. In some hypertensive persons, a postprandial natriuretic response, normally elicited by a salty meal, is diminished and contributes to body sodium accumulation and plasma volume expansion. An important physiological mechanism ensuring the increased salt excretion following ingestion of salt is based on a luminocrine and endocrine secretion of novel small intestinal peptides--guanylins. Membrane guanylate cyclase receptors mediate effects of these peptides that provide a novel link between the intestine and kidney by means of circulating molecular guanylin forms. It can be expected that the emergence of the novel guanylin signaling pathways will energize search for molecular defects causing hypertension. PMID- 11409352 TI - [The autonomic nervous system and hypertension]. AB - It was shown that dysregulation of the cardiovascular system plays an important role in the development of arterial hypertension. Sympathetic drive can be increased mainly in cases of borderline and essential hypertension. The rise in sympathetic activity is not well balanced by the adequate increase in parasympathetic activity, even the PS tone is diminished. Relative sympathotony can occur beside the absolute sympathotony due to the decrease in the PS tone with normal or quantitatively less diminished sympathetic drive. The cause of the ANS activity dysbalance remains unknown. Probably there exist disorders of generator and modulator activities in higher levels of CNS, baroreceptor regulation and other peripheral (humoral and other) mechanisms. Adequate and appropriate recording of different cardiovascular parameters mainly during activation of ANS by maneuvers (orthostasis, Valsalva, etc.) is necessary. Beat to-beat registrations of blood pressure (e.g. by Penaz method FINAPRES), heart rate (RR intervals) are necessary for the evaluation of short term blood pressure and heart rate fluctuations. Important information about S/PS activities and the balance in heart rate regulation can be achieved by spectral and non-linear analysis of the HR variability. Regular physical aerobic exercise increases the parasympathetic tone and is beneficial for restoration of the disordered S/PS balance in young hypertonics. It can be used as non-pharmacological treatment of arterial hypertension. PMID- 11409353 TI - [Role of the GABA-ergic and adrenergic system in transmission and modulation of pain and possibilities of pharmacologic control]. AB - The dorsal horn of the spinal cord contains many transmitters and receptors involving in pain transmission and modulation. Monoamines and gamma-aminobutyric acid (GABA) play a key role in the modulation at level of dorsal horn (DH) of the spinal cord, the site of primary processing of afferent nociceptive information. Agents that enhance the action of GABA at the GABAA receptors can produce antinociception. For example intrathecal administration of benzodiazepines increases pain threshold in various models of pain. There is also substantial evidence for robust antinociceptive properties of spinal administration of clonidine and other alpha 2-agonists. PMID- 11409354 TI - [The most frequently cited publications of Czech pharmacologists]. AB - The most cited articles of 100 Czech pharmacologists were determined by analysis of data given in the Science Citation Index in "Web of Science". Ten most cited articles were selected according to the total number of citations till October 2000, the total number of citations during 1980-2000 and the number of citations per year during 1980-2000. The most cited articles found by this analysis represent well the significant part of research orientation of Czech pharmacologists and their major scientific contributions during the 1960s-1990s. Problems with application of citation-analysis to evaluation of research are discussed. PMID- 11409355 TI - [Nobel Prize winning laureates in physiology or medicine for the year 2000--a few comments on discoveries related to signal transduction]. AB - The Nobel Assembly awarded The Nobel Prize in Physiology or Medicine for 2000 jointly to Arvid Carlsson, Paul Greengard and Eric Kandel for their discoveries concerning signal transduction in the nervous system. On the examples of their predecessors we attempted to demonstrate how results of basic research serve as building blocks for new discoveries and for the application of research results into the praxis. We presented not only the basic discoveries of laureates of Nobel Prize for year 2000 (biological role of dopamine, regulation of cell functions by phosphorylation of proteins, changes in transduction of signals during processes of memory), but we also mentioned previous discoveries that helped in the research of the last laureates. These discoveries concerned not only the storage and metabolism of transmitters, formulation of the concept of cyclic AMP as a second messenger of hormonal action, the role of G-proteins in transduction processes in receptor-effector complexes, processes of phosphorylation of proteins as regulators of cell functions, but we also mentioned the discovery of other second messengers and substances functioning as local hormones (prostaglandins and related compounds). Most of the described discoveries have not only the value as stones that can help to fill still incomplete mosaic of our present knowledge, but they also represent the immediate basis for the development and use of very important remedies, such as are antiparkinsonics, antidepressive drugs, nonsteroidal antiinflammatory drugs, etc. PMID- 11409356 TI - Web alert. PMID- 11409357 TI - Web alert. PMID- 11409358 TI - Web alert. PMID- 11409362 TI - [Report on XIII International AIDS Conference, Durban, South Africa--the Durban Declaration by the scientists and a speech on AIDS and poverty by the President of South Africa]. PMID- 11409363 TI - [An old time on my life in research work]. PMID- 11409364 TI - [Report on 25th International Herpesvirus Workshop]. PMID- 11409365 TI - [Impressions on 11th International Conference on Negative Strand Viruses]. PMID- 11409366 TI - [Report on XIII International AIDS Conference, Durban, South Africa]. PMID- 11409367 TI - [Report on Options for the Control of Influenza Virus IV]. PMID- 11409368 TI - [Impressions of VIIIth Parvovirus Workshop]. PMID- 11409369 TI - [The 101st annual meeting of the Japan Surgical Society. Sendai, Japan. April 11 13, 2001. Abstracts]. PMID- 11409370 TI - [75th annual meeting of the Japanese Association for Infectious Diseases. Nara, Japan. March 29-30, 2001. Abstracts]. PMID- 11409371 TI - [The 41st annual meeting of the Japanese Respiratory Society. Tokyo, Japan. April 4-6, 2001. Abstracts]. PMID- 11409372 TI - The (non-)sense of the methionine loading test for detecting hyperhomocysteinemia is not ready for a compromise. PMID- 11409373 TI - [The 105th annual meeting of the Japanese Ophthalmological Society. Yokohama, Japan. April 19-21, 2001. Abstracts]. PMID- 11409374 TI - [The 45th Congress of the Japan Rheumatism Association. Tokyo, Japan. May 14-16, 2001. Abstracts]. PMID- 11409375 TI - The prognostic value of myocardial viability recognized by low dose dipyridamole echocardiography in patients with chronic ischaemic left ventricular dysfunction. AB - AIMS: The aim of this study was to assess the prognostic value of myocardial viability recognized as a contractile response to vasodilator stimulation in patients with left ventricular dysfunction in a large scale, prospective, multicentre, observational study. METHODS AND RESULTS: Three hundred and seven patients (mean age 60 +/- 10 years) with angiographically proven coronary artery disease, previous (>3 months) myocardial infarction and severe left ventricular dysfunction (ejection fraction <35%; mean ejection fraction: 28 +/- 7%) were enrolled in the study. Each patient underwent low dose dipyridamole echo (0.28 mg x kg(-1) in 4 min). Myocardial viability was identified as an improvement of >0.20 in the wall motion score index. By selection, all patients were followed up for a median of 36 months. One-hundred and twenty-four were revascularized either by coronary artery bypass grafting (n=83) or coronary angioplasty (n=41). The only end-point analysed was cardiac death. In the revascularized group, cardiac death occurred in one of the 41 patients with and in 16 of the 83 patients without a viable myocardium (2.4% vs 19.3%, P<0.01). Outcome, as estimated by Kaplan-Meier survival, was better for patients with, compared to patients without, a viable myocardium, who underwent coronary revascularization (97.6 vs 77.4%, P=0.01). Using a Cox proportional hazards model, the presence of myocardial viability was shown to exert a protective effect on survival (chi square 4.6, hazard ratio 0.1, 95% CI 0.01-0.8, P<0.03). The survival rate in medically treated patients was lower than in revascularized patients irrespective of the presence of a viable myocardium (79.7% vs 86.2, P=ns). CONCLUSION: In severe left ventricular ischaemic dysfunction, myocardial viability, as assessed by low dose dipyridamole echo, is associated with improved survival in revascularized patients. PMID- 11409376 TI - Differences in acetate recovery factor between groups may interfere with tracer estimates of fact oxidation. PMID- 11409377 TI - An appreciation of Elvin A. Kabat (1914-2000): scientist, educator, and a founder of modern carbohydrate biology. PMID- 11409378 TI - Reiter's disease following Mycoplasma pneumoniae infection. PMID- 11409379 TI - Optimal treatment regimens for patients with bleeding disorders. PMID- 11409380 TI - Leechcraft. PMID- 11409381 TI - Relationship between nonphenacetin-combined analgesics and nephropathy. PMID- 11409382 TI - Relationship between nonphenacetin-combined analgesics and nephropathy. PMID- 11409383 TI - Relationship between nonphenacetin-combined analgesics and nephropathy. PMID- 11409384 TI - [Computers in dental research and practice]. PMID- 11409385 TI - [Question of the month (catgut sutures)]. PMID- 11409386 TI - [Question of the month: Antibiotic prophylaxis for dental patients with joint replacements]. PMID- 11409387 TI - The real cost of aspirin. PMID- 11409388 TI - Hyperthyroidism induced by beta-human chorionic gonadotrophin. PMID- 11409389 TI - The problem of conduction velocity of the human spinothalamic tract. PMID- 11409390 TI - Tobacco extract used as a remedy for urinary retention 150 years ago by the native population of the Balkans. PMID- 11409393 TI - New patient per radiation oncologist annual workload: comment. PMID- 11409394 TI - Radiotherapy utilization: reply. PMID- 11409395 TI - Rectal perforation during double contrast barium enema. PMID- 11409396 TI - Why are doctors so unhappy? Healing and happiness go together. PMID- 11409397 TI - Why are doctors so unhappy? Doctors have conceded their autonomy. PMID- 11409400 TI - Risk of miscarriage in pregnant users of NSAIDs. More information is needed to be able to interpret study's results. PMID- 11409401 TI - Risk of miscarriage in pregnant users of NSAIDs. Miscarriages also occur in women intending to have induced abortions. PMID- 11409402 TI - Quality of health information about depression on internet. Study's shortcomings may have affected findings. PMID- 11409403 TI - Quality of health information about depression on internet. Level of evidence should be gold standard. PMID- 11409404 TI - Nasal diamorphine in children with clinical fractures. Patients should be told what to do when analgesia wears off. PMID- 11409405 TI - Nasal diamorphine in children with clinical fractures. Most interesting questions remain unanswered in this study. PMID- 11409406 TI - Patient access to records must be acceptable to both parties. PMID- 11409407 TI - Computer use must not affect doctor-patient relationship. PMID- 11409408 TI - Unlimited consumer demand would destroy NHS. PMID- 11409409 TI - 2E4/Kaptin (KPTN)--a candidate gene for the hearing loss locus, DFNA4. AB - Stereocilia of the inner ear play an integral role in the mechanotransduction of sound. Their structural support is derived from actin filaments and actin-binding proteins. We have identified a novel actin-binding protein, 2E4-kaptin (KPTN), which appears to be involved in this structural network. Using double label immunofluorescence, we now show that KPTN extends beyond the barbed ends of actin filaments at the tips of stereocilia, and using cloned human cDNA, we mapped KPTN to chromosome 19q13.4. A combination of FISH, radiation hybrid mapping and YAC screening localized KPTN between markers D19S412 and NIB1805, making this gene an excellent functional and positional candidate for DFNA4, a form of autosomal dominant non-syndromic hearing loss. We identified a second family with inherited deafness that also maps to the DFNA4 region. To screen KPTN for deafness-causing mutations, we first determined its genomic structure and then completed a mutational analysis by direct sequencing and SSCP in affected family members. Although no deafness-causing mutations were identified in the coding region, KPTN remains an excellent candidate gene for hearing loss; by synteny, its murine orthologue also remains a candidate gene for the Nijmegan waltzer (nv) mouse mutant, which has vestibular defects and a variable sensorineural hearing loss. PMID- 11409410 TI - Search for multifactorial disease susceptibility genes in founder populations. AB - The current challenge in biomedical research is to detect genetic risk factors involved in common complex diseases. The power to detect their role is generally poor in populations that have been large for a long time. It has been suggested that the power may be increased by taking advantage of the specificity of founder populations: linkage disequilibrium spanning larger regions and kinship coefficients being stronger than in large populations. A new method is proposed here, the Maximum Identity Length Contrast (MILC) which, in contrast with other existing methods, does not make the assumption of unique ancestry for the genetic risk factors. It is thus appropriate for a search for common genetic risk factors for complex diseases. Statistical properties of the method are discussed in realistic contexts. PMID- 11409411 TI - Laparoscopic intragastric resection of gastric leiomyoma using needlescopic instruments. Case report. AB - Laparoscopic intragastric resection of gastric leiomyoma was performed using needlescopic instruments. The patient was a 71-year-old man who had a 2-year history of gastric submucosal tumor 2 cm in diameter located near the esophagocardiac junction. After getting informed consent, we performed a laparoscopic intragastric tumor resection under an oral endoscope. There were no intra- or postoperative complications. The patient was discharged uneventfully. Histopathologic diagnosis of the tumor was leiomyoma. Laparoscopic intragastric resection of a benign gastric submucosal tumor using needlescopic instruments is technically feasible and as safe as a less invasive procedure. PMID- 11409412 TI - The minimally invasive surgical suite enters the 21st century. A discussion of critical design elements. AB - Most minimally invasive surgical procedures are now performed in operating rooms that were originally designed for traditional open surgery. Laparoscopic instrumentation such as insufflators, light sources, and camera control units must be placed on one or more equipment carts. After the cart has been moved into place, insufflation tubing, video cables, light cords, cautery lines, and foot controls must be positioned and connected. This cart-based paradigm restricts the ergonomic configuration of the operating room and creates potential mechanical, electrical, and biological hazards to the patient and operating room staff. In order to decrease clutter, ease personnel movement, improve ergonomics, maintain the sterile field, and facilitate the use of advanced imaging, communication, and display devices, an appropriately designed operating environment is essential. Herein we detail both the theoretical and practical aspects of the design and describe the implementation and utilization of such a suite in our hospital. These design elements may prove to be critical to the next generation of minimally invasive surgical suites and will facilitate future advanced laparoscopic procedures. PMID- 11409413 TI - The clinical and diagnostic implications of mosaicism in the neurofibromatoses. AB - Neurofibromatosis type 1 and type 2 both occur in mosaic forms. Mosaicism results from somatic mutations. Early somatic mutations cause generalized disease, clinically indistinguishable from nonmosaic forms. Later somatic mutation gives rise to localized disease often described as segmental. In individuals with mosaic or localized manifestations of neurofibromatosis type 1 (segmental neurofibromatosis type 1), disease features are limited to the affected area, which varies from a narrow strip to one quadrant and occasionally to one half of the body. Distribution is usually unilateral but can be bilateral, either in a symmetric or asymmetrical arrangement. Patients with localized neurofibromatosis type 2 have disease-related tumors localized to one part of the nervous system; for example a unilateral vestibular schwannoma with ipsilateral meningiomas or multiple schwannomas in one part of the peripheral nervous system. The recognition of mosaic phenotypes is important. Individuals with the mosaic form, even with a generalized phenotype, are less likely to have severe disease. They also have lower offspring recurrence risk than individuals with the nonmosaic form. The mosaic forms of neurofibromatosis provide a good example of the effects of somatic mutation. It is increasingly recognized that mild and unusual forms of many dominantly inherited disorders are caused by the same mechanism. PMID- 11409414 TI - Clinical neurophysiology training and certification in the United States: 2000. PMID- 11409415 TI - Pyridostigmine-induced microcephaly. PMID- 11409416 TI - Radiographic joint damage in rheumatoid arthritis: a community-based perspective. PMID- 11409417 TI - Cerebral amyloid angiopathy: a microvascular link between parenchymal and vascular dementia? PMID- 11409418 TI - When running a stop sign may be a good thing. PMID- 11409419 TI - Variants of the Guillain Barre syndrome: progress toward fulfilling "Koch's postulates". PMID- 11409420 TI - Novel amyloid precursor protein mutation in an Iowa family with dementia and severe cerebral amyloid angiopathy. AB - Several mutations in the amyloid precursor protein (APP) gene have been found to associate with pathologic deposition of the beta-amyloid peptide (Abeta) in neuritic plaques or in the walls of cerebral vessels. We report a mutation at a novel site in APP in a three-generation Iowa family with autosomal dominant dementia beginning in the sixth or seventh decade of life. The proband and an affected brother had progressive aphasic dementia, leukoencephalopathy, and occipital calcifications. Neuropathological examination of the proband revealed severe cerebral amyloid angiopathy, widespread neurofibrillary tangles, and unusually extensive distribution of Abeta40 in plaques. The affected brothers shared a missense mutation in APP, resulting in substitution of asparagine for aspartic acid at position 694. This site corresponds to residue 23 of Abeta, thus differing from familial Alzheimer's disease mutations, which occur outside the Abeta sequence. Restriction enzyme analysis of DNA from 94 unrelated patients with sporadic cerebral amyloid angiopathy-related hemorrhage found no other instances of this mutation. These results suggest a novel site within Abeta that may promote its deposition and toxicity. PMID- 11409421 TI - Gentamicin treatment of Duchenne and Becker muscular dystrophy due to nonsense mutations. AB - Aminoglycosides have previously been shown to suppress nonsense mutations, allowing translation of full-length proteins in vitro and in animal models. In the mdx mouse, where muscular dystrophy is due to a nonsense mutation in the dystrophin gene, gentamicin suppressed truncation of the protein and ameliorated the phenotype. A subset of patients with Duchenne and Becker muscular dystrophy similarly possess a nonsense mutation, causing premature termination of dystrophin translation. Four such patients, with various stop codon sequences, were treated once daily with intravenous gentamicin at 7.5 mg/kg/day for 2 weeks. No ototoxicity or nephrotoxicity was detected. Full-length dystrophin was not detected in pre- and post-treatment muscle biopsies. PMID- 11409422 TI - Animal model of axonal Guillain-Barre syndrome induced by sensitization with GM1 ganglioside. AB - Some humans develop the axonal form of Guillain-Barre syndrome after receiving bovine brain ganglioside. On sensitization with the ganglioside mixture, all of a group of rabbits injected developed high anti-GM1 IgG antibody titers, flaccid limb weakness of acute onset, and a monophasic illness course. Pathological findings for the peripheral nerves showed predominant Wallerian-like degeneration, with neither lymphocytic infiltration nor demyelination. IgG was deposited on the axons of the anterior roots, and GM1 was proved to be present on the axons of peripheral nerves. Sensitization with purified GM1 also induced axonal neuropathy, indicating that GM1 was the immunogen in the mixture. A model of human axonal Guillain-Barre syndrome has been established that uses inoculation with a bovine brain ganglioside mixture or isolated GM1. This model may help to clarify the molecular pathogenesis of the syndrome and to develop new treatments for it. PMID- 11409423 TI - Focal cooling rapidly terminates experimental neocortical seizures. AB - The efficacy of surgical resection for epilepsy is considerably lower for neocortical epilepsy than for temporal lobe epilepsy. We have explored focal cooling with a thermoelectric (Peltier) device as a potential therapy for neocortical epilepsy. After creating a cranial window in anesthetized rats, we induced seizures by injecting artificial cerebrospinal fluid containing 4 aminopyridine (4-AP), a potassium channel blocker. Within 30 minutes of 4-AP injection, animals developed recurrent seizures (duration 85.7 +/- 26.2 seconds; n = 10 rats) that persisted for 2 hours. When a small Peltier device cooled the exposed cortical surface to 20-25 degrees C at seizure onset, the seizure duration was reduced to 8.4 +/- 5.0 seconds (n = 10 rats; p < 0.001). When the Peltier device was placed close to the cortical surface, but not allowed to make physical contact, there was no effect on seizure duration (104.3 +/- 20.7 seconds; p > 0.05 compared to control). Interestingly, the duration of uncooled seizures was reduced after we allowed the cortex to rewarm from prior cooling. Histological examination of the cortex after cooling has shown no evidence of acute or delayed neuronal injury, and blood pressure and temperature remained stable. It may be possible to use Peltier devices for cortical mapping or, when seizure detection algorithms improve, for chronic seizure control. PMID- 11409424 TI - Adenosine A2A receptor activation reduces proinflammatory events and decreases cell death following intracerebral hemorrhage. AB - The ubiquitous neuromodulator adenosine inhibits the production of several proinflammatory cytokines through activation of specific cell-surface adenosine receptors. We demonstrated recently that antisense oligonucleotides to tumor necrosis factor-alpha (TNF-alpha) are neuroprotective in a rat model of intracerebral hemorrhage. Therefore, we hypothesized that activation of adenosine receptors would provide protection against intracerebral hemorrhage-induced TNF alpha production and inflammatory events. In vitro experiments showed that adenosine A1, A2A, and A3 receptor subtypes were present on U937 cells, and activation of these subtypes inhibited TNF-alpha production with a rank order of A2A > > A1 > A3. Prolonged treatment of U937 cells with the A2A receptor agonist 2-p-(carboxyethyl)phenethylamino-5'-N-ethylcarboxamidoadenosine hydrochloride (CGS 21680) desensitized adenosine A2A, A1, and A3 receptors. CGS 21680 administration directly into the striatum immediately prior to the induction of intracerebral hemorrhage inhibited TNF-alpha mRNA and, 24 hours following induction, reduced parenchymal neutrophil infiltration (p < 0.001) and TUNEL positive cells (p < 0.002) within and bordering the hematoma. These results suggest that pharmacological strategies targeting A2A receptors may provide effective inhibition of acute neurotoxic proinflammatory events that occur following intracerebral hemorrhage. PMID- 11409425 TI - Apolipoprotein E phenotype and the efficacy of intravenous tissue plasminogen activator in acute ischemic stroke. AB - We used stored plasma samples from 409 patients in the National Institute of Neurological Diseases and Stroke (NINDS) tissue plasminogen activator (t-PA) Stroke Trial to examine the relationship between an apolipoprotein (Apo) E2 or an Apo E4 phenotype and a favorable outcome 3 months after stroke, the risk of intracerebral hemorrhage, and the response to intravenous t-PA therapy. For the 27 patients with an Apo E2 phenotype who were treated with t-PA, the odds ratio (OR) of a favorable outcome at 3 months was 6.4 [95% confidence interval (CI) 2.7 15.3%] compared to the 161 patients without an Apo E2 phenotype who were treated with placebo. The 190 patients treated with t-PA who did not have an Apo E2 phenotype also had a greater, though less pronounced, likelihood of a favorable outcome (OR 2.0, 95% CI 1.2-3.2%) than patients without an Apo E2 phenotype treated with placebo. For the 31 patients with an Apo E2 phenotype treated with placebo, the OR of a favorable 3 month outcome was 0.8 (95% CI 0.4-1.7%) compared to the 161 patients without an Apo E2 phenotype treated with placebo. This interaction between treatment and Apo E2 status persisted after adjustment for baseline variables previously associated with 3 month outcome, for differences in the baseline variables in the two treatment groups and in the Apo E2-positive and -negative groups, and for a previously reported time-to-treatment x treatment interaction (p = 0.03). Apo E4 phenotype, present in 111 (27%) of the 409 patients, was not related to a favorable 3 month outcome, response to t-PA, 3 month mortality, or risk of intracerebral hemorrhage. We conclude that the efficacy of intravenous t-PA in patients with acute ischemic stroke may be enhanced in patients who have an Apo E2 phenotype, whereas the Apo E2 phenotype alone is not associated with a detectable benefit on stroke outcome at 3 months in patients not given t-PA. In contrast to prior studies of head injury and stroke, we could not detect a relationship between Apo E4 phenotype and clinical outcome. PMID- 11409426 TI - Correlation between neurological progression and astrocyte apoptosis in HIV associated dementia. AB - The pathogenesis of HIV-associated dementia (HIVD) has been postulated to be due to the indirect effects of HIV infection, including the aberrant central nervous system production of cytokines and other neurotoxins. A correlation between the severity of dementia and production of neurotoxins in HIVD has been demonstrated. We have previously identified nonproductive HIV infection of astrocytes. Because astrocytes participate in the inactivation of neurotoxins, we hypothesize that HIV nonproductive infection of astrocytes may lead to an environment in which there is a significant level of astrocyte apoptosis and a consequent increase in the levels of neurotoxins and that this results in more rapidly progressing dementia. Postmortem brain tissue was examined from clinically well-characterized HIV-positive demented patients, HIV-positive nondemented patients, and HIV seronegative nondemented control subjects. The HIVD group was further categorized into subjects with rapid and those with slow progression of dementia. Tissue was paraformaldehyde fixed and paraffin embedded, and 6-microm sections from the basal ganglia and mid-frontal gyrus were processed to detect apoptosis by in situ transferase dUTP nick end labeling. Astrocytes were co-localized using immunohistochemical techniques. In situ polymerase chain reaction (PCR) techniques were utilized to detect HIV DNA in astrocytes. The density of apoptotic astrocytes was significantly greater in the HIV-positive groups than in the HIV-negative group (p < 0.01). The HIVD rapid progressors had a significantly greater number of apoptotic astrocytes in the basal ganglia than did the HIVD slow progressors (p < 0.05). In addition, there were a greater number of HIV DNA positive astrocytes, as demonstrated by in situ PCR, in the HIVD rapid progressors than in the slow progressor and HIV-nondemented groups. These data suggest that there is an increased rate of astrocyte loss in the subjects with rapidly progressive dementia, in association with an increased number of HIV DNA positive astrocytes. The results emphasize the importance of understanding more completely the role of HIV infection of astrocytes in the neuropathogenesis of HIVD. PMID- 11409427 TI - Delayed cerebral edema and fatal coma after minor head trauma: role of the CACNA1A calcium channel subunit gene and relationship with familial hemiplegic migraine. AB - Trivial head trauma may be complicated by severe, sometimes even fatal, cerebral edema and coma occurring after a lucid interval ("delayed cerebral edema"). Attacks of familial hemiplegic migraine (FHM) can be triggered by minor head trauma and are sometimes accompanied by coma. Mutations in the CACNA1A calcium channel subunit gene on chromosome 19 are associated with a wide spectrum of mutation-specific episodic and chronic neurological disorders, including FHM with or without coma. We investigated the role of the CACNA1A gene in three subjects with delayed cerebral edema. Two subjects originated from a family with extreme FHM, and one subject was the previously asymptomatic daughter of a sporadic patient with hemiplegic migraine attacks. In all three subjects with delayed severe edema, we found a C-to-T substitution resulting in the substitution of serine for lysine at codon 218 (S218L) in the CACNA1A gene. The mutation was absent in nonaffected family members and 152 control individuals. Haplotype analysis excluded a common founder for both families. Neuropathological examination in one subject showed Purkinje cell loss with relative preservation of granule cells and sparing of the dentate and inferior olivary nuclei. We conclude that the novel S218L mutation in the CACNA1A calcium channel subunit gene is involved in FHM and delayed fatal cerebral edema and coma after minor head trauma. This finding may have important implications for the understanding and treatment of this dramatic syndrome. PMID- 11409428 TI - Risk of tremor and impairment from tremor in relatives of patients with essential tremor: a community-based family study. AB - Essential tremor (ET) is a common condition that is present in as many as 23% of elderly individuals. Our objective was to determine the risk of ET and to study the impairment resulting from ET among relatives of ET cases compared to relatives of controls. ET cases and matched controls from the Washington Heights Inwood community, New York, and their first- and second-degree relatives underwent a standardized tremor examination. The risk of having ET in relatives of cases vs relatives of controls was compared using Cox proportional hazards models. Five hundred ninety-one subjects were examined (59 ET cases, 72 controls, 234 case relatives, and 226 control relatives). ET was present in 25 (22.5%) of the 111 first-degree relatives of cases compared to 6 (5.6%) of 107 first-degree relatives of controls [relative risk (RR) = 4.67, 95% confidence interval (CI) = 1.90-11.49, p = 0.0008]. RRs were higher in relatives of cases with onset < or =50 years than in those with later onset (RR = 10.38 vs 4.82). Sixteen (64%) of twenty-five affected first-degree case relatives exhibited moderate tremor while performing tasks such as writing, drinking, or pouring. Relatives of ET patients are five times more likely to develop the disease than are members of the population and ten times more likely if the proband's tremor began at an early age. The majority of the affected relatives can expect to experience impairment resulting from tremor. PMID- 11409429 TI - Texture analysis and morphological processing of magnetic resonance imaging assist detection of focal cortical dysplasia in extra-temporal partial epilepsy. AB - In many patients, focal cortical dysplasia (FCD) is characterized by minor structural changes that may go unrecognized by standard radiological analysis. To increase the sensitivity of magnetic resonance imaging (MRI) for the detection of subtle lesions of FCD, we developed voxel-based image postprocessing methods, including first-order texture analysis and morphological processing modeled on known MRI features of FCD. We selected 16 patients with histologically proven FCD. Image processing features were calculated over a neighborhood for each voxel in the three-dimensional T1-weighted MRI. Three feature maps were generated: (1) gray matter thickness map to model cortical thickening, (2) gradient map to model blurring of the gray matter-white matter junction, and (3) relative intensity map to model the hyperintense signal within the lesion. Two observers detected lesions on conventional MRI in 8/16 and on ratio maps in 14/16 patients. Sensitivity was 87.5% (14/16) for the ratio maps compared to 50% (8/16) for MRI (p < 0.003). Specificity was 95% (19/20) for ratio maps and 100% (20/20) for MRIs. Cohen's kappa was 0.53 for MRIs, indicating moderate agreement, and 0.83 for ratio maps, indicating strong agreement beyond chance between the 2 observers. The image-processing methods developed in this study improve visual detection of FCD, even in cases where no lesion is obvious on MRI. These techniques could increase the number of patients with partial epilepsy who could benefit from surgery. PMID- 11409430 TI - Platelet-derived growth factor-alpha receptor-positive oligodendroglia are frequent in multiple sclerosis lesions. AB - Platelet-derived growth factor (PDGF) ligand is a potent glial cell mitogen. When its cognate receptor (PDGF-alphaR) is expressed on oligodendroglial lineage cells, such cells are considered capable of division, and the receptor thus serves as a phenotypic marker for oligodendrocyte precursor cells. Here we identify using immunohistochemistry a considerably enlarged, PDGF-alphaR expressing oligodendrocyte cell population within multiple sclerosis (MS) white matter lesions compared to control brains. Numerous PDGF-alphaR-positive oligodendroglia also colabel heavily with the nuclear cell proliferation marker antibody Ki-67. Our finding of large numbers of proliferating oligodendroglia in MS brains expressing up-regulated PDGF-alphaR suggests that these progenitor-like cells represent an important source of regenerating cells for the healing MS lesion. PMID- 11409431 TI - Evidence for digenic inheritance in a family with both febrile convulsions and temporal lobe epilepsy implicating chromosomes 18qter and 1q25-q31. AB - We report a clinical and genetic study of a French family among whom febrile convulsions (FC) are associated with subsequent temporal lobe epilepsy (TLE) in the same individual, without magnetic resonance imaging-identifiable hippocampal abnormalities. Linkage analyses excluded the loci FEB1 and FEB2, previously implicated in FC; the GEFS+1 locus responsible for generalized epilepsy with febrile seizures plus; and the locus implicated in lateral temporal lobe epilepsy. After scanning the entire genome, significant lod scores (>3) for markers on 18qter and suggestive lod scores (>2) for markers on 1q25-q31 were obtained. An analysis of the haplotypes at these two loci supported the hypothesis that two genes segregated with the phenotype. All patients shared common haplotypes for both 1q25-q31 and 18qter chromosomes. All but one unaffected at-risk individuals carried only one, or none, of the disease haplotypes. Under the assumption of digenic inheritance, haplotype reconstruction defined a 26 cM interval on chromosome 1 and a 10 cM interval on chromosome 18. This family suggests that the association between FC and TLE may be observed in the absence of hippocampal structural abnormalities and that they may have, in some cases, a common genetic basis. PMID- 11409432 TI - A longitudinal MRI study of histopathologically defined hypointense multiple sclerosis lesions. AB - Severe tissue destruction is the presumed histopathological correlate of hypointense multiple sclerosis (MS) lesions. In this study we correlated changes of lesion hypointensity over time with initial histopathological features in 14 biopsied MS lesions. The extent of hypointensity increased in initially demyelinated plaques and decreased in remyelinating lesions. The initial axonal loss determined the increase of hypointensity over time. In conclusion, both axonal loss and demyelinating activity determine the evolution of hypointensity over time. PMID- 11409433 TI - A SURF1 gene mutation presenting as isolated leukodystrophy. AB - Mitochondrial respiratory chain defects are increasingly recognized in patients with leukodystrophy. We report the first case of leukodystrophy with systemic cytochrome oxidase deficiency caused by a loss of function mutation in the SURF1 gene in a 2-year-old girl presenting with failure to thrive, global neurodevelopmental regression, and lactic acidosis. Although all previously reported mutations in the SURF1 gene have been found in patients with cytochrome oxidase (COX)-deficient Leigh syndrome, the phenotype associated with SURF1 protein deficiency should be extended to include leukodystrophy. PMID- 11409434 TI - Slowing of voluntary and involuntary saccades: an early sign in spinocerebellar ataxia type 7. AB - We describe quantitative oculomotor findings in a patient with subclinical spinocerebellar ataxia type 7 (SCA7) and a borderline mutation of 38 CAG repeats and her daughter with SCA7 and 46 repeats. Both subjects demonstrated significant slowing of voluntary and involuntary saccades, but retinal examination was normal. Smooth pursuit and fixation suppression of VOR were mildly impaired. Slow saccades may be the earliest neurologic finding even in asymptomatic SCA7 patients with normal ocular fundi. The SCA7 mutation probably has an early impact on brainstem fast eye movement centers. PMID- 11409435 TI - Intermediate CAG repeat lengths (53,54) for MJD/SCA3 are associated with an abnormal phenotype. AB - We report on a Dutch family in which 4 members in 2 generations have intermediate repeat lengths (53 and 54) for Machado-Joseph Disease/Spinocerebellar Ataxia (MJD/SCA3). All but the youngest have a restless legs syndrome with fasciculations and a sensorimotor axonal polyneuropathy. Central neurological abnormalities are only present in 2. This family shows that intermediate repeat lengths can be pathogenic and may predispose for restless legs and peripheral nerve disorder. PMID- 11409436 TI - Autoantibodies to amyloid-beta and Alzheimer's disease. AB - Immunization against amyloid-beta has been suggested as a possible preventive or therapeutic treatment for Alzheimer's disease. We hypothesized that some individuals may have autoantibodies to amyloid-beta and that this may be protective. We analyzed the plasma of 365 individuals, drawn from a larger longitudinal epidemiological study, for the presence of antibodies to amyloid beta. There were detectable but very low levels of anti-amyloid-beta antibodies in just over 50% of all samples and modest levels in under 5% of all samples. However, neither the presence nor the level of anti-amyloid-beta antibodies correlated with the likelihood of developing dementia or with plasma levels of amyloid-beta peptide. These data suggest that low levels of anti-amyloid-beta autoantibodies are frequent in the elderly population but do not confer protection against developing dementia. PMID- 11409437 TI - Treatment of acute Nipah encephalitis with ribavirin. AB - Nipah virus, a newly identified paramyxovirus caused a severe outbreak of encephalitis in Malaysia with high fatalities. We report an open-label trial of ribavirin in 140 patients, with 54 patients who were managed prior to the availability of ribavirin or refused treatment as control. There were 45 deaths (32%) in the ribavirin arm; 29 deaths (54%) occurred in the control arm. This represents a 36% reduction in mortality (p = 0.011). There was no associated serious side effect. This study suggests that ribavirin is able to reduce the mortality of acute Nipah encephalitis. PMID- 11409438 TI - Right lateralized motor cortex activation during volitional blinking. AB - Using H2 15O positron emission tomography in 6 healthy volunteers, we found that self-initiated and externally cued blinking activated the right primary motor cortex and supplementary motor area (SMA). The left dorsolateral prefrontal cortex (DLPFC) and the rostral SMA showed greater activation during the self initiated task compared to the externally cued task. This study confirms the hypothesis of right hemispheric lateralization of volitional blinking derived from observations in stroke patients. Furthermore, it underscores the role of DLPFC and rostral SMA in self-initiated movements, which has been found in similar experiments with hand movements. PMID- 11409441 TI - Lack of association of the interleukin-1alpha gene polymorphism with Alzheimer's disease in a Korean population. PMID- 11409442 TI - Gene polymorphisms of interleukin-1alpha influence the course of Alzheimer's disease. PMID- 11409443 TI - Low cerebral blood flow velocity and risk of white matter hyperintensities. PMID- 11409444 TI - [The 43rd annual meeting of the Japanese Society of Child Neurology. Okayama, Japan. June 7-9, 2001. Program and Abstracts]. PMID- 11409445 TI - Biomechanical properties after pre-twist of canine patellar tendon. AB - We studied what effect a rotational pre-twist of the patellar tendon had on its mechanical properties. We used the central strip of canine patellar tendons. The length, width and thickness of each specimen were measured and the specimens were mounted in custom-made aluminum pots. Five groups of 10 specimens each were measured with neutral rotation, 90 degrees external rotation, 180 degrees external rotation, 270 degrees external rotation and 360 degrees external rotation. The ultimate stress, ultimate strain, average elastic modulus, and strain energy density were measured. There were no statistically significant differences among the five groups. Twisting the graft up to 360 degrees did not decrease the biomechanical properties of the graft. PMID- 11409446 TI - Ceramic endoprosthesis for aggressive giant cell tumors near the knee: eight cases followed for more than 10 years. AB - Eight patients with locally aggressive giant cell tumor were treated with marginal resection and implantation of a ceramic endoprosthesis with a non-hinged knee joint. The patients have been followed-up for between 10 to 16 years. Newly formed bone appeared at the junction between implant and cortex and in 2 cases subsidence of the implant was seen. All the patients had good function of the knee and no revision was needed. PMID- 11409447 TI - The Saddle prosthesis in periacetabular tumours. AB - The Saddle endoprosthesis provides a means of establishing a stable and mobile articulation between the femur and a partially resected pelvis. Six patients with malignant or aggressive benign bone tumours underwent resection and replacement with custom-made Saddle endoprosthesis. Wide margin was achieved in four cases and marginal margin in two. Follow-up ranged from 24 to 41 months. All the six patients were then alive, five being disease-free. One patient developed deep infection and local recurrence necessitating removal of the prosthesis. The functional results were either excellent or good in five patients. PMID- 11409448 TI - Treatment of low-energy tibial shaft fractures: plaster cast compared with intramedullary nailing. AB - We analyzed data from 87 patients who had displaced closed or open grade I simple or spiral wedge tibial shaft fractures caused by low-energy impact. Fifty-four patients were treated with plaster cast and 33 with intramedullary locking nail (IMLN). Delayed union only occurred in 8 patients after plaster cast treatment. Forty-two patients in the IMLN group and one in the plaster cast group suffered from anterior knee pain. Final treatment outcome, healing time, hospitalization time and duration of sick leave were assessed on the basis of 25 matched pairs of patients. Mean healing time, hospitalization time and sick leave in the plaster cast and IMLN groups were 19 (SD 6.7) and 12 (SD 4.4) weeks (P<0.001); 8 (SD 4.8) and 7 (SD 2.7) days (P=0.686); and 195 (SD 81) and 106 (SD 31) days (P=0.001), respectively. No difference was found between plaster cast and IMLN groups when the outcome was evaluated using the criteria of Johner and Wruhs. PMID- 11409449 TI - Long-term results of tibialis posterior tendon transfer for drop-foot. AB - Twelve patients with drop-foot secondary to sciatic or common peroneal nerve palsy treated with transfer of the tibialis posterior tendon were followed-up for a mean of 90 (24-300) months. In 10 patients the results were 'excellent' or 'good'. In 11 patients grade 4 or 5 power of dorsiflexion was achieved, although the torque, as measured with a Cybex II dynamometer, and generated by the transferred tendon, was only about 30% of the normal side. Seven patients were able to dorsiflex their foot to the neutral position and beyond. The results appeared to be better in men under 30 years of age with common peroneal palsies. A painful flatfoot acquired in adulthood does not appear to be a significant long term complication despite the loss of a functioning tibialis posterior tendon. PMID- 11409450 TI - Osteoid osteoma: the results of surgical treatment. AB - Between 1987 and 1999 we diagnosed and treated 110 patients with osteoid osteoma. Sixty patients were younger than 20 years. One hundred and four patients had characteristic pain at night. The mean duration of symptoms before surgery was 16 months. One hundred and four symptomatic patients were treated operatively with either wide resection or curettage. Ninety-one patients had immediate and complete relief of pain. The average follow-up was 2.5 years. PMID- 11409451 TI - Primary lymphoma of bones. AB - Primary malignant lymphoma of bone (PLB) is an uncommon tumour. A survey of 278 primary malignant cases in our clinic showed that 20 cases of PLB were registered between 1986 and 1997. Fourteen of the 20 cases underwent surgical excision. The mean follow-up time was 36.3 months. The rate of response to treatment was 65% with a rate of complete recovery of 55%. The clinical stage of tumours correlated well with the prognosis. Our results seem worse than most of the series in the literature. A high proportion of stage IV disease and pathological fractures at presentation may be a factor. PMID- 11409452 TI - Chronic osteomyelitis: a continuing orthopaedic challenge in developing countries. AB - Nine patients with chronic osteomyelitis, three with problems due to diagnosis, three with dilemmas regarding treatment and three with other complications are presented. It is suggested that although the diagnosis of osteomyelitis in most cases is straightforward, presentation might sometimes be similar to other conditions, which can lead to a dilemma in the diagnosis. Because of the formidable complications, which may be difficult to manage and the difficulty in guaranteeing permanent cure, the best approach is prevention by judicious treatment of acute haematogenous osteomyelitis and of open fractures. PMID- 11409453 TI - Scoliosis curve correction, thoracic volume changes, and thoracic diameters in scoliotic patients after anterior and after posterior instrumentation. AB - Thoracic volume was calculated in 50 adolescent patients operated on for severe idiopathic thoracic scoliosis. In 25, anterior instrumentation was used (group 1), and posterior instrumentation in the other 25 patients (group 2). Calculation of thoracic volume was made from measurements of pre-operative and post-operative radiographs. The mean spinal curvature in group 1 was 73+/-12.4 degrees before the operation, and 19+/-15 degrees after the operation, and in group 2 the curvature was 75+/-13 degrees before the operation and 37+/-10 degrees after the operation. The calculated thoracic volume in the group with anterior instrumentation increased from 5234 ml pre-operatively to 6043 ml post operatively, while with posterior instrumentation it increased from 5155 ml to 5489 ml. The correlation between the change in the Cobb angle and the thoracic volume change was poor for both groups. To determine the role in the thoracic volume increase of the frontal, sagittal and vertical thoracic diameters, further correlation tests were made between these and the thoracic volume increase in each diameter. The best correlation was found between the frontal and vertical increase of diameters in group 1, whereas in group 2 the best correlation was found between the volume increase and the sagittal parameters. PMID- 11409454 TI - Surgical correction of Scheuermann's kyphosis. AB - This is a retrospective study of eight consecutive patients of mean age 19 (13 27) years with severe Scheuermann's kyphosis who underwent anterior and/or posterior fusion using the Cotrel-Dubousset (CD) instrumentation. In two an anterior release and fusion with rib grafts had been previously performed. The mean follow-up was 5 years. The preoperative hyperkyphosis averaged 86 degrees (71 degrees - 99 degrees), which was postoperatively 44 degrees (32 degrees - 58 degrees). The average loss of correction was 4.6 degrees (1 degrees - 12 degrees). The lumbar hyperlordosis spontaneously improved from -67 degrees to -48 degrees. Two patients, who had chronic back pain refractory to conservative treatment, improved considerably after surgery. PMID- 11409455 TI - Aspergillus spondylitis in immunocompetent patients. AB - Four immunocompetent patients with neurological deficit underwent anterior decompression for Aspergillus osteomyelitis of the spine. All patients improved neurologically following anterior spinal decompression and antifungal therapy. This study emphasizes the importance of obtaining a tissue diagnosis as these unusual infections may mimic tuberculosis, which is more common. PMID- 11409456 TI - Increased loosening of cemented straight stem prostheses made from titanium alloys. An analysis and comparison with prostheses made of cobalt-chromium-nickel alloy. AB - We studied the rate of aseptic loosening of three different types of femoral stems in primary total hip replacement. After a median follow-up of 10.2 years 4/147 CoCrNi (SS77) straight stems (type M.E. Muller) were revised. After a median follow-up of 7.7 years 32/239 Ti-6A1-7Nb (SS77) were revised and after a median follow-up of 5.2 years 52/203 SLS Titanium alloy stems were revised. Whereas the first two stems are of identical design (smooth-blasted, anterior and posterior collar), the SLS stem design is different. Surgical procedure and cementing technique have remained unchanged. There is a significantly higher risk of failure for smaller titanium stem sizes and in males and patients who are physically active. This indicates that the greater elasticity of the Titanium alloy is one of the factors responsible for loosening. PMID- 11409457 TI - Results of a cemented titanium alloy straight femoral shaft prosthesis after 10 years of follow-up. AB - Two-hundred fifty implantations of a cemented femoral stem made of titanium alloy in 239 patients were followed for 9.7 years (range 8.7-10.3 years). Eighty-nine patients with 93 hips have died and two could not be located. Five hips have been revised, two for infection, one for aseptic loosening and two during revision of the cup. Three stems showed radiological loosening but have not been revised. The average hip score was 85. The results are encouraging and comparable to other cemented femoral stems. PMID- 11409458 TI - A proximal fixed anatomic femoral stem reduces stress shielding. AB - In 24 patients with total hip replacement using a short anatomic femoral stem, bone mineral density (BMD) was measured after a 7-year follow-up using dual energy X-ray absorptiometry. The contralateral side was used as a control. The BMD on the side of the prosthesis was lower by a mean of 7% than that on the control side. The difference was greatest in the area of the calcar and laterally and proximally around the stem. BMD at the metaphyseal and diaphyseal areas were the same as on the contralateral side. The bone loss around the proximal aspect of the stem may be related to the proximal porous coating. It is concluded that stress shielding can be diminished by appropriate design of the femoral component. PMID- 11409459 TI - Periprosthetic bone mineral density changes with femoral components of differing design philosophy. AB - We measured the bone mineral density in 22 patients with the cylindrical stemmed cobalt-chrome AML prosthesis (collared) and in 22 patients with the tapered stem titanium CLS prosthesis (collarless). DEXA scanning was undertaken at a mean of 40 months in the AML and 52 months in the CLS group from the time of implant insertion. In both groups the greatest mean loss of BMD was found in Gruen zone 7 and the least change in Gruen zone 5. In all zones the BMD loss was greater in the AML group but only statistically significant in zones 6 (P<0.05) and 7 (P<0.01). Although numerous factors affect BMD changes around cementless implants, this study suggests that less bone loss can be associated with the titanium CLS stem. PMID- 11409460 TI - High tibial osteotomy for valgus and varus deformities of the knee. AB - In 53 patients with a mean age of 38 (17-73) years, 71 high tibial osteotomies were performed. Twenty-three patients had no radiological signs of osteoarthrosis (Ahlback grade 0), whereas 26 patients had primary and 22 patients secondary osteoarthritis. Follow-up was 10.5 (5.8-16.6) years. The patients without radiological osteoarthrosis achieved almost exclusively good or very good scores using the Lysholm-Gillquist (96%) and the Insall scale (83%). By contrast, only 29% of the patients with radiological osteoarthritis achieved good or very good scores. Whether the patient had primary or secondary osteoarthrosis was of no influence. Neither was the preoperative axial deviation. PMID- 11409461 TI - Should the patella be replaced in total knee replacement? AB - In 170 total knee arthroplasties for osteoarthritis 71 did not receive a patellar replacement (group A), while 99 knees had a cemented polyethylene patella (group B). The mean follow-up time was 36 months (30-50 months). In group A 10 patients underwent second-stage patellar resurfacing and in group B 2 knees underwent revision of the patellar component. Radiologically the average patellar congruency was similar. In both groups there were 21 non-congruent knees. In group A 8 were symptomatic and had low scores compared to 2 in group B (P<0.05). The mean HSS score and patellar score were higher in group B than in group A (P<0.05). PMID- 11409462 TI - Ligand-receptor interaction under periodic stimulation: a modeling study of concentration chemoreceptors. AB - The first step of chemosensory transduction consists in the association of ligand molecules with receptor proteins borne by the cell membrane. In this article, the time evolution of ligand-receptor complexes is studied in the presence of a periodically changing ligand concentration. This type of stimulation is a close approximation to some natural situations, for example in olfaction. The transient and steady-state periodic levels of the complexes, resulting from a single-step (binding) or double-step (binding and activation) reaction, are determined. When possible, analytical solutions are given, if not for the complete model, at least for its simplified version at low ligand concentration. Otherwise, solutions are found numerically and both the complete and simplified versions of the model are compared. The results obtained are discussed with respect to actual experimental data based on the moth sex-pheromone receptor. Periodic steady states are achieved very quickly and their amplitude decreases when the stimulation frequency increases. We show that the simplified description is adequate if only a fraction of activated receptors is sufficient to produce the maximum response, as is actually the case in the example treated. The role of the frequency of stimulation is investigated and it is shown to possess an optimal range between 2 and 5 Hz. PMID- 11409463 TI - Modification of anomalous swelling in multilamellar vesicles induced by alkali halide salts. AB - By use of small-angle X-ray scattering it is shown that addition of alkali halide salts in small amounts (0-200 mM) shifts the repeat spacing in multilamellar DC13PC vesicles and alters the anomalous swelling behaviour close to the main transition. Both effects follow the Hofmeister series of the ions. We suggest that the shift of repeat spacing can be explained by ion effects on the van der Waals attractive forces between the membranes and on the decay length of the repulsive hydration force. The anomalous swelling is explained in terms of a critical unbinding of the membranes. The proximity of the critical temperature of the unbinding to the main transition temperature can be tuned by varying the concentration and type of salt in the sample. PMID- 11409464 TI - Brownian dynamics simulations of fluorescence fluctuation spectroscopy. AB - We have developed a program for the simulation of the fluorescence fluctuations as detected from highly diluted samples of (bio)molecules. The model is applied to translational diffusion and takes into account the hydrodynamic interactions. The solution concentration is kept constant by assuming periodic boundary conditions and spans here the range 0.5< C < 10 nM. We show that the fluorescence correlation functions can be accurately computed on systems of limited size (a few molecules per simulation box) by simulating for a total time approximately 100-300 times the diffusion relaxation time of the fluorescence autocorrelation function. The model is applied also to the simulation of the scanning fluorescence correlation spectroscopy (FCS) and of the photon counting histograms for the confocal collection configuration. Scanning FCS simulations of highly diluted samples (C approximately equals 0.5 nM) show anticorrelation effects in the autocorrelation functions of the fluorescence signal that are less evident for higher concentrations. We suggest here that this effect may be due to the non uniform occupancy of the scanning area by the fluorophores. PMID- 11409465 TI - Buffer effects on electric signals of light-excited bacteriorhodopsin mutants. AB - The effects of glycyl-glycine and bis-trispropane buffers on the light-excited electric signals due to proton motion in the molecule were studied for the bacteriorhodopsin (bR) mutants D38R, D96N, E204Q, R227Q, D85N, D85T, R82Q/D85N, and D85N/D96N in purple membranes and for delipidated purple membrane containing the wild-type bR. The results show additional charge motion caused by the buffers in all cases. Arrhenius parameters calculated from the temperature dependence of the difference signals (with buffer minus without buffer) are similar to the parameters found for the wild-type bR in the case of these buffers: the values of the activation enthalpies are mostly in the range 25-50 kJ/mol; all the activation entropies are negative. The results are evaluated with the cluster hypothesis outlined previously. PMID- 11409466 TI - Dynamics and orientation of amphipathic peptides in solution and bound to membranes: a steady-state and time-resolved fluorescence study of staphylococcal delta-toxin and its synthetic analogues. AB - The environment of both the hydrophilic and hydrophobic sides of alpha-helical delta-toxin are probed by tryptophanyl (Trp) fluorescence, when self-association occurs in solution and on binding to membranes. The fluorescence parameters of staphylococcal delta-toxin (Trp15 on the polar side of the amphipathic helix) and synthetic analogues with single Trp at position 5 or 16 (on the apolar side) were studied. The time-resolved fluorescence decays of the peptides in solution show that the local environment of their single Trp is always heterogeneous. Although the self-association degree increases with concentration, as shown by fluorescence anisotropy decays, the lifetimes (and their statistical weight) of Trp16 do not change, contrary to what is observed for Trp15. The first step of self-association is then driven by hydrophobic interactions between apolar sides of alpha-helices, whilst further oligomerization involves their polar side (Trp15) via electrostatic interactions. This is supported by dissociation induced by salt. For all self-associated peptides, the polarity of the Trp microenvironment was not significantly modified upon binding to phospholipid vesicles, as indicated by the small shifts of the fluorescence emission spectra and lifetime values. However, the relative populations of the lifetime classes vary with bound-peptide density similar to the rates of their global motions in bilayers or smaller particles. Quenching experiments by water or lipid-soluble compounds show changes of the orientation of membrane-inserted peptides, from probably dimers lying flat at the interface at low peptide density, to oligomers spanning the membrane and inducing membrane fragmentation at high peptide density. PMID- 11409467 TI - Atomic force pulling: probing the local elasticity of the cell membrane. AB - We present a novel approach, based on atomic force microscopy, for exploring the local elastic properties of the membrane-skeleton complex in living cells. Three major elements constitute the basis for the proposed method: (1) pulling the cell membrane by increasing the adhesion of the tip to the cell surface provided via appropriate tip modification; (2) measuring force-distance curves with emphasis on selecting the appropriate withdrawal regions for analysis; (3) fitting of the theoretical model for axisymmetric bending of an annular thick plate to the experimental curve in the withdrawal region, prior to the detachment point of the tip from the cell membrane. This approach, applied to human erythrocytes, suggests a complimentary technique to the commonly used methods. The local use of this methodology for determining the bending modulus of the cell membrane of the human erythrocyte yields a value of (2.07+/-0.32) x 10(-19) J. PMID- 11409468 TI - Short-range specific forces are able to induce hemifusion. AB - Working with pure lipidic systems (giant unilamellar vesicles, 10-150 microm in diameter) as models for biological membranes, we have considered possible structures of the contact area of two adherent membranes by investigating the diffusion of fluorescent lipid analogues from one vesicle to another. Two bilayers in close contact can almost be seen as a lamellar structure in equilibrium. This is the usual configuration of two adherent vesicles, in which the interbilayer distance is estimated to be 3 nm. We have increased the attraction between the membranes by either adding depletion forces or by using a trick, inspired from the interaction between nucleic bases in nucleosides (herein adenosine and thymidine). The nucleosides were attached to the polar head of amphiphilic molecules that behave like phospholipids and were incorporated in the model membrane. The extra attraction between two membranes, resulting from base pairing, strongly decreased the interbilayer distance down to about 1 nm. This change of the water content induced lipid rearrangements, which could also be viewed in terms of a phase transition at low water content. These rearrangements were not observed in the case of depletion forces. We conclude that the introduction of an additional attractive force in the system modifies the equilibrium state, leading to a drastic change in the membrane behavior, which will tentatively be related to hemifusion. PMID- 11409469 TI - 4',6-diamidino-2-phenylindole (DAPI) interacts with rare structures of GC polymers. AB - The binding of 4',6-diamidino-2-phenylindole (DAPI) to double-stranded GC polymers either in the alternating or in homopolymer sequence was investigated using fluorescence techniques. We employed fluctuation correlation spectroscopy, which measures the diffusion coefficient of fluorescent particles, to demonstrate that the fluorescence was originating from relatively slowly diffusing entities. These entities display a very large heterogeneity of diffusing coefficients, indicating that molecular aggregation is extensive in our samples. We used frequency domain fluorometry to characterize the fluorescence lifetime of the species, while varying the GC polymer-dye coverage systematically. At very low coverage we observed a relatively bright fluorescent component with a lifetime value of approximately 4 ns. The stoichiometry of binding of this bright species was such that it can only arise from rare molecular structures, either unusual loops or large molecular aggregates. The amount and characteristics of this bright fluorescent component were different between the homo and the alternating polymer, indicating that the difference in sequence of the two polymers is responsible for the different aggregates which are then detected in the fluorescence experiment. At large GC polymer coverage we observed a relatively wide distribution of fluorescent species with short lifetime values, in the range between 0.12 and 0.2 ns. Given the stoichiometry of binding of this fluorescent component, we concluded that it could arise either from intercalative and/or non specific binding to the DNA double-stranded molecules. We comment on the origin of the rare but brightly fluorescent binding sites and discuss the potential to detect such unusual DNA structures. PMID- 11409470 TI - Placenta accreta associated with a ruptured pregnant rudimentary uterine horn. Case report and review of the literature. AB - Pregnancy in a rudimentary uterine horn is rare and is usually associated with fetal death and serious maternal morbidity and mortality. A case of pregnancy in a rudimentary uterine horn with rupture 14 weeks after last menstrual period and is complicated with placenta accreta is presented. The patient had signs and symptoms of massive hemoperitoneum. An emergency exploratory laparotomy revealed rupture of the gravid rudimentary horn of a bicornuate uterus. Histologic examination of the specimen showed that placenta was accreta. The relative literature is reviewed and the association of placenta accreta in such situations is pointed out. PMID- 11409471 TI - Complete migration of retained surgical sponge into ileum without sign of open intestinal wall. AB - A 24-year-old woman came to the emergency room with a history of diffuse abdominal pain in the form of colic, nausea, vomiting and intestinal constipation. Clinical and ultrasound findings suggested intestinal obstruction due to foreign body. She had been submitted to a cesarean section 4 months previously at another hospital. At laparotomy, a ileum loop was found to be distended by an inside large and hardened mass with another intestinal loops and omentum density adherent. An ileotomy was performed on the compromised segment with terminating anastomosis. When opened surgical specimen it was observed an intraluminal surgical sponge that had completely migrated into the interior of the ileum and stopped next to ileumcecal valve. No fistulas or open intestinal wall were observed. PMID- 11409472 TI - Vaginal leiomyoma co-existing with broad ligament and multiple uterine leiomyomas. AB - A 48 year old African American woman presented with bladder pressure leading to the diagnosis of broad ligament and multiple uterine leiomyomas. She was also found to have a lateral vaginal wall mass which was confirmed to be a leiomyoma. Unlike uterine leiomyomas, vaginal leiomyomas are uncommon and are most often found in Caucasian women. Cases of such coexisting tumors are rare and their etiologic relationship is uncertain. PMID- 11409473 TI - Primary non-Hodgkin's lymphoma of the uterine cervix. AB - We present a patient with primary malignant lymphoma of the uterine cervix, which is extremely rare. She had a complete remission with a combination of chemotherapy and radiotherapy. Measurement of the serum soluble interleukin-2 receptor level was a useful index of the effectiveness of treatment. PMID- 11409474 TI - Incidence of intrauterine complications in monoamniotic twin gestation. AB - We compared the incidence of intrauterine complications, such as twin-twin transfusion syndrome, between monochorionic-monoamniotic (MM) and monochorionic diamniotic (MD) twin gestations. We retrospectively evaluated a series of 11 MM and 211 MD twin gestations managed after 22 weeks' gestation at our hospitals. One MM twin gestation was diagnosed with twin-twin transfusion syndrome based on clinical presentation of polyhydramnios and hydrops. The incidence of twin-twin transfusion syndrome in MM twin gestations (9.1%) was not measurable different from that in the MD twin gestations (7.6%, p=0.09). There were no measurable differences in the incidence of other complications, such as growth discordance >25% or intrauterine fetal demise between the two groups. MM twin gestations should be kept under close management while carefully monitoring for any circulatory imbalance between the twins. PMID- 11409475 TI - Extra-strong graduated compression stocking reduces usage of vasopressor agents during spinal anesthesia for cesarean section. AB - Because a standard type of graduated compression stocking is not effective for the prevention of spinal anesthesia hypothension during cesarean section, we have used an extra-strong type of graduated compression stocking. This study examined whether the extra-strong stocking reduces usage of vasopressor agents. Forty eight and 47 full term parturients were fitted with the standard stocking and the extra-strong stocking, respectively. Spinal anesthesia was performed by injecting 2.0 ml 0.3% dibucaine hyperbaric solution. When systolic blood pressure decreased to 90-100 mmHg or to less than 70% of the pre-anesthesia value, ephedrine was injected. There was no significant difference in systolic blood pressure or heart rate during spinal anesthesia between the groups. The mean dose of ephedrine injected during anesthesia was 12.2 and 4.3 mg, and the incidence of ephedrine injected was 85 and 49%, in the standard and extra-strong stocking groups, respectively. The extra-strong stocking group showed significantly lower values. Fitting the extra-strong stocking on both legs for cesarean sections undergoing spinal anesthesia reduced usage of vasopressor agents, suggesting a promising use of this stocking for non-invasive prophylaxis. PMID- 11409476 TI - Laser therapy in chronic cervicitis. AB - Chronic cervicitis is one of the common disorders encountered in daily practice. To overcome this problem first step should be antibiotic therapy in acute phase. If this fails, the infection becomes chronic and may spread to internal genital organs leading to pelvic inflammatory disease and eventually to infertility. Chronic form of infection may necessitate tissue destruction to prevent the development of dysplasia and neoplasm. Various methods such as electrocautery, loop diathermy, cryotherapy or laser are used to destroy the inflamed area. In this study we present 26 cases of chronic cervicitis managed with Nd-YAG laser with a success rate of 93%. We advice this method to block the development of cervical intraepithelial neoplasm. PMID- 11409477 TI - The role of ovarian volume in an in vitro fertilization programme as assessed by 3D ultrasound. AB - The study was designed to investigate the role of ovarian volume, as assessed by three-dimensional (3D) sonography, in predicting conception in an in-vitro fertilization-embryo transfer (IVF-ET) programme. Transvaginal 3D sonography was performed in 152 cycles before initiation of ovarian stimulation (day 1) and on the day of oocyte retrieval. Ovarian volume showed no significant correlation with IVF outcome. On the contrary, all ovarian measurements were lower, albeit nonsignificantly, in the conception group. Fifteen patients (15/152, 9.9%) had a minimum unilateral ovarian volume of < or =3 mL (1 SD below the mean) on day 1 of the stimulation cycle. In this subgroup, the likelihood of conception was 6.7% (1/15) versus 21.9% (30/137) in patients with an initial minimum ovarian volume of >3 mL. This difference did not reach statistical significance. In both groups, cancellation rates due to poor ovarian response or lack of fertilization were similar. In conclusion, ovarian volumetry as assessed by three-dimensional ultrasound failed to predict conception in women undergoing IVF treatment. PMID- 11409478 TI - A study of prolidase in mothers, their newborn babies and in non-pregnant controls. AB - The prolidase activity in serum and in erythrocytes was measured in 28 healthy mothers and in the cord blood of their newborn babies in using a modified Chinard method. 45 healthy non-pregnant women aged between 15-36 years formed a control group. Biochemical parameters (CK, BUN, C-peptid, AFP, Uric acid) were also measured. The serum and erythrocyte prolidase activities in maternal blood were 45.8+/-13.4 U/L and 37.8+/-2.7 U/g Hb respectively. There was no significant difference in the enzyme activities between pregnant women and the control group (p>0.05). However serum and erythrocyte prolidase activity in cord blood (20.3+/ 8.2 U/L and 31.6+/-7.3 U/g Hb) was significantly different when compared with control group (53.4+/-14.7 U/L in serum and 42.3+/-10.3 U/g Hb in erythrocyte, p<0.001). There was a significant correlation between maternal and cord blood serum enzyme activity (r: 0.76 p<0.01). This correlation was also shown in erythrocyte prolidase activities of both groups (r: 0.49, p<0.05). Cord blood prolidase activity was positively correlated with birth weight (r: 0.89, p<0.01). Prolidase activity in cord blood was low even though collagen turnover is increased in fetal growth. PMID- 11409479 TI - Pregnancy outcome and birth spacing among Palestinian women between 1940 and 1989. AB - In a sample of 1,250 Palestinian women living in cities, villages and refugee camps of the West Bank over a period of 50 years, we studied the number of pregnancies, children and miscarriages per woman, the waiting time to the first pregnancy, birth spacing between children, and the average age of women at first live birth. The average number of pregnancies, the percentage of miscarriages, the waiting time to the first pregnancy and the birth interval between children decreased with time in all locations. The average age of women at first live birth in cities and villages showed very little difference, but slightly lower in refugee camps. PMID- 11409480 TI - Methotrexate versus hyperosomolar glucose in the treatment of extrauterine pregnancy. AB - The aim of this prospective, randomized, double blind study was to compare the efficacy of methotrexate and hyperosmolar glucose injected directly into the extra-uterine gestational sac under laparoscopic vision. The study included twenty women with ectopic pregnancy. Inclusion criteria were intact tubal pregnancy, not exceeding 4 cm in diameter, rising or plateauing betahCG levels, and no evidence of intra-abdominal bleeding. The patients were treated by laparoscopically guided injection of 3 mL fluid into the area containing the tubal pregnancy. The fluid contained either 25 mg methotrexate (n=9) or 50% glucose (n=9). Daily decrease in betahCG levels was faster in patients treated by methotrexate (median 8.7%) than in those treated by hyperosmolar glucose (median 4.8%), p=0.17. The study was discontinued due to a higher failure rate in the group treated by hyperosmolar glucose. In conclusion, local injection of methotrexate is superior to hyperosmolar glucose. It can be used as an alternative to salpingostomy or salpingotomy whenever laparoscopy is performed for the diagnosis and treatment of extra-uterine pregnancy. PMID- 11409481 TI - Natural history of endometrial hyperplasia. Study of 77 patients. AB - Seventy-seven patients with endometrial hyperplasia, 48 with simple hyperplasia without atypia (SH), 17 with complex hyperplasia without atypia (CH), one with simple hyperplasia with atypia (SHA), and 11 with complex hyperplasia with atypia (CHA) were prospectively followed-up by total curettage every 12 months for 3 years. Progression to carcinoma occurred in only one of the 77 patients; she showed grade 1 adenocarcinoma. The overall regression rates were 79% for SH, 100% for SHA, 94% for CH, and 55% for CHA, respectively. In patients with CHA whose disease reverted to normal endometrium, regression was most likely to occur within the first year. PMID- 11409482 TI - Fetal survival despite unrecognized uterine rupture resulting from previous unknown corporeal scar. AB - Cesarean scar rupture of a gravid uterus with unknown corporeal scar is common. Our case was a 35 year woman, gravida 2, para 1 presented at 38 weeks gestation. She was admitted to our hospital for routine follow up. She had no signs or symptoms of labor. However eight hours after the initial examination, she came back to hospital with the signs of shock and acute abdomen. Immediately she was referred to surgery. Intraoperatively a complete rupture of the classical corporeal incision was observed, but the fetus was enclosed within the anterior lying plasenta. The fetus was delivered with one minute apgar score 3, and five minute apgar score 8. According to this case, we conclude that spontaneous uterine rupture of the classical uterine scar can be observed even without uterine contractions. So women with the possibility of previous classical uterine incision should be delivered once fetal maturity is documented. PMID- 11409483 TI - Ovarian lymphomas. A clinicopathological analysis of 10 cases. AB - Analysis of 10 cases of Non-Hodgkin's lymphoma with initial manifestation in the ovaries is presented with the histologic and immunohistochemical findings. PMID- 11409484 TI - Leiomyoma of the vesicovaginal septum. AB - A 69-year-old woman presented with a solid homogenous mass in the area of the posterior wall of the urinary bladder. A transvaginal approach was used to remove the mass located in the vesicovaginal septum. Histology revealed a benign smooth spindle cell tumor and immunohistochemistry confirmed leiomyoma. PMID- 11409485 TI - Magnetic resonance imaging of a benign Brenner tumor with an ipsilateral simple cyst. AB - Magnetic resonance imaging features were assessed in a benign Brenner tumor with an ipsilateral simple cyst. Brenner tumor showed isointensity to the uterine muscle on T1-weighted images and markedly low signal intensity on T2-weighted images. On the other hand, on T1-weighted images after administration of gadopentetate dimeglumine a Brenner tumor showed mild patchy enhancement. The low signal intensity seen on T1- and T2-weighted images may have been due to the abundant dense fibrous tissues. PMID- 11409487 TI - Eicosanoid production by human monocytes: does COX-2 contribute to a self limiting inflammatory response? AB - The eicosanoids, prostaglandin E2 (PGE2) and thromboxane A2 (TXA2), are involved in inflammatory events. TXA2 has potentially pro-inflammatory actions and PGE2 has actions which can be considered both pro- and antiinflammatory. Therefore, it is potentially significant that production of TXA2 and PGE2 by stimulated monocytes have very different time courses. TXA2 synthesis is immediate and dependent on cyclooxygenase Type 1 (COX-1) activity whereas PGE2 synthesis is delayed and dependent on COX-2 activity. These apparent COX-isotype dependencies of TXA2 and PGE2 synthesis can be explained by differences in the affinities of TXA synthase and PGE synthase for the common substrate, PGH2. The findings have implications for the use of NSAIDs and selective COX-2 inhibitors whose actions can increase the monocyte TXA2/PGE2 ratio. PMID- 11409486 TI - Granulocyte-colony stimulating factor in the prevention of postoperative infectious complications and sub-optimal recovery from operation in patients with colorectal cancer and increased preoperative risk (ASA 3 and 4). Protocol of a controlled clinical trial developed by consensus of an international study group. Part three: individual patient, complication algorithm and quality manage. AB - GENERAL DESIGN: Presentation of a new type of a study protocol for evaluation of the effectiveness of an immune modifier (rhG-CSF, filgrastim): prevention of postoperative infectious complications and of sub-optimal recovery from operation in patients with colorectal cancer and increased preoperative risk (ASA 3 and 4). A randomised, placebo controlled, double-blinded, single-centre study is performed at an University Hospital (n = 40 patients for each group). This part presents the course of the individual patient and a complication algorithm for the management of anastomotic leakage and quality management. OBJECTIVE: In part three of the protocol, the three major sections include: The course of the individual patient using a comprehensive graphic display, including the perioperative period, hospital stay and post discharge outcome. A center based clinical practice guideline for the management of the most important postoperative complication--anastomotic leakage--including evidence based support for each step of the algorithm. Data management, ethics and organisational structure. CONCLUSIONS: Future studies with immune modifiers will also fail if not better structured (reduction of variance) to achieve uniform patient management in a complex clinical scenario. This new type of a single-centre trial aims to reduce the gap between animal experiments and clinical trials or--if it fails--at least demonstrates new ways for explaining the failures. PMID- 11409488 TI - The inflammatory response in humans after inhalation of bacterial endotoxin: a review. AB - There is increasing evidence that diseases caused by organic dusts are mainly of an inflammatory nature. Among the many agents present in organic dusts, bacterial endotoxin is a major candidate for the inflammatory reaction. The purpose of this paper was to review the inflammatory response in humans after inhalation of bacterial endotoxin (lipopolysaccharide, LPS) in order to improve the understanding of symptoms and reactions found among persons exposed to endotoxin containing organic dusts. It has been reported that inhalation of LPS causes changes in forced expiratory volume in one second (FEV1), and forced vital capacity (FVC). At the alveolar level, inhalation of LPS can induce changes in the diffusion capacity. Activation and migration of neutrophils are major effects of acute LPS inhalation. Changes in mediators of inflammation, such as eosinophilic cationic protein (ECP), myeloperoxidase (MPO), interleukin-8 (IL-8), IL-1beta, tumor necrosis factor alpha (TNFalpha) and C-reactive protein (CRP) in the airways and/or blood, have also been found. Inhalation of 30-40 microg LPS seems to be a threshold level for inducing clinical symptoms and lung function changes in healthy subjects. The threshold dose for inducing changes in blood neutrophils may be less than 0.5 microg LPS. In conclusion, available data regarding the responses to LPS inhalation challenges demonstrate a local and a systemic inflammatory response at lower doses of LPS, while higher inhaled doses are required to elicit significant clinical and lung function responses. Future inhalation studies on LPS need to focus on relevant diagnostic tools for the inflammatory reaction among persons exposed to endotoxin-containing organic dusts and to evaluate whether the large variation between individuals in the response to organic dusts or endotoxin could be due to differences in the molecular mechanisms responsible for the toxicity of the agent. PMID- 11409489 TI - In-vitro test system for the evaluation of cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2) inhibitors based on a single HPLC run with UV detection using bovine aortic coronary endothelial cells (BAECs). AB - OBJECTIVE AND DESIGN: The aim of this study was to develop a new, whole-cell test system which is easy to handle and requires a standard equipment for the parallel screening of COX-1 and COX-2 inhibitors. MATERIALS: Bovine aortic endothelial cells (BAECs). TREATMENT AND METHODS: Unstimulated bovine aortic coronary endothelial cells (BAECs) were used as a source of COX-1 and BAECs pretreated with ASA (100 microM) and activated with phorbol myristate acetate (PMA) were used as a source of COX-2. The time- and concentration-dependent induction of COX 2 expression in the BAECs was evaluated by a kinetic profile (HPLC analysis) and detected by Western-Blot analysis using polyclonal antibodies against COX-1 and COX-2. RESULTS: In BAECs, diclofenac and meloxicam showed balanced inhibition of COX-1 (IC50: 0.01/0.4 microM) and COX-2 (IC50: 0.03/0.6 microM). Indomethacin inhibited COX-1 more potently than COX-2 (IC50: 0.008/0.04 microM). Aceclofenac inhibited COX-2 more potently than COX-1 (IC50: 3.0/7.3 microM). DFU and Cl SC57666 [16] inhibited COX-2 (IC50: 0.04/0.001 microM) highly selectively but did not inhibit COX-1 (IC50: >100 microM). CONCLUSIONS: In summary an assay has been developed, for the determination of IC50-values for inhibitors of COX-1/2 on cells of the same origin, in line with values in the literature. Moreover, new insights have been gained into the relationship of COX-1/2 and lipoxygenase pathways in BAECs by detecting 15- and 12-HETE: Inhibition of COX-1 by the NSAIDs mostly resulted in an enhancement of 15-HETE and 12-HETE release. In contrast inhibition of COX-2 decreased 15-HETE release. PMID- 11409490 TI - Basic fibroblast growth factor up-regulates the surface expression of complement receptors on human monocytes. AB - OBJECTIVE AND DESIGN: To clarify the possible involvement of basic fibroblast growth factor (b-FGF) in inflammation, we examined the effect of b-FGF on the surface expression of complement receptors (CR) on human monocytes in vitro. MATERIALS AND METHODS: Heparinized venous blood was obtained from healthy adult donors. The surface expression of CR on blood monocytes was determined by two color immunofluorescent staining using flow cytometry and monoclonal antibodies. A standard whole blood lysis technique was used to avoid any in vitro manipulation that would activate monocytes. RESULTS: b-FGF increased the expression of CR3 on monocytes in a dose- and time-dependent manner. The b-FGF concentrations used were up to 100 ng/ml. The values of mean fluorescence intensity (MFI) of CR3 expression on unstimulated monocytes were 12.6+/-1.3 (n = 3), whereas those on b-FGF-stimulated monocytes were 59.2+/-7.1 (n = 3). b-FGF also up-regulated the expression of CR1 on monocytes in a dose- and time dependent manner. The MFI values of CR1 expression on unstimulated monocytes were 2.5+/-0.1 (n = 3), whereas those on b-FGF-stimulated monocytes were 11.1+/-0.6 (n=3). The magnitude of CR1 expression by monocytes was significantly smaller than that of CR3 expression. The maximal stimulatory effect of b-FGF on monocytes was observed using greater than 25 ng/ml of b-FGF and 90-120 min incubation period. CONCLUSION: b-FGF may participate in the inflammatory process by modulating the CR expression on blood monocytes. PMID- 11409491 TI - Sensitisation of group III articular afferents to mechanical stimuli by substance P. AB - OBJECTIVE: To examine the excitatory and sensitising effects of substance P (SP) on articular afferents in normal and acutely inflamed cat knee joints. MATERIALS AND METHODS: In anesthetised cats recordings were made from 15 group III (conduction velocity 2.5-20 m/s) and 25 group IV afferent units (conduction velocity < 2.5 m/s) of the medial articular nerve of normal and acutely inflamed knee joints. SP (10 and 100 microg) was administered close-arterially. RESULTS: SP at doses of 10 microg and 100 microg activated less than 50% of both group III and group IV units. The proportion of SP-positive units was significantly higher in inflamed (10 of 21) than in normal joints (2 of 18). SP induced activity in initially silent units or increased ongoing activity after latencies varying from 2 s to 5 min. The SP-evoked activity had an irregular pattern, a variable duration, and was not related to the dose injected. Bolus injections of SP (100 microg) sensitised group III articular afferents but not group IV units to noxious movements of the joint, regardless whether the units were from normal or acutely inflamed joints. The responses to innocuous movements were not influenced by SP. Group III units, initially not activated by any movement, displayed vigorous discharges to noxious movements after close-arterial SP. In 3 group III units tested, the SP-induced augmentation of responses to noxious movements was not mimicked by close-arterial injection of histamine (3.3 microg). CONCLUSIONS: It is concluded that SP contributes to the sensitisation of a subpopulation of high-threshold articular afferents. Thus this neuronal mediator released peripherally in response to an injury or acute inflammation causes considerable changes in the mechanosensitivity of this subpopulation of nociceptive joint afferents. PMID- 11409492 TI - Participation of cyclooxygenases in cutaneous thermal nociception under non inflamed and inflamed conditions. AB - OBJECTIVE AND DESIGN: Effects of cyclooxygenase inhibitors on noxious thermal stimuli were investigated in non-inflamed and inflamed rats. MATERIALS: Male Sprague-Dawley rats were used in this study. TREATMENT: Cyclooxygenase inhibitors, indomethacin, mofezolac, NS-398, and JTE-522 were administered orally at a dose of 10 mg/kg 1 h prior to and 4 h after the intravenous injection of lipopolysaccharide (1 mg/kg). METHODS: The nociceptive response was evaluated from the escape latency of foot withdrawal to the thermal stimuli with a beam of light. Expression ofcyclooxygenase was examined by reverse transcription polymerase chain reaction. RESULTS: In normal rat, administration of indomethacin, relatively cyclooxygenase-1-selective inhibitor, mofezolac, or cyclooxygenase-2-selective inhibitors, NS-398 and JTE-522 had no effects on the escape latency against thermal stimuli. Injection of lipopolysaccharide into rat induced the expression of mRNA for cyclooxygenase-2 in the subcutaneous tissue of foot pad. The escape latency at 8 h was significantly shortened by the injection. This hyperalgesia could be reversed by pretreatment of rat with NS-398 or JTE 522, but not with mofezolac. CONCLUSIONS: Cyclooxygenases may have little participation in peripheral skin thermal nociception in non-inflamed condition, although cyclooxygenase-2 could be responsible for the hyperalgesia during inflammation induced by lipopolysaccharide. PMID- 11409493 TI - Interaction of imidazolinone herbicides with soil humic acids. Experimental results and molecular modeling. AB - Adsorption and desorption isotherms of the herbicides imazapyr, imazethapyr and imazaquin on a soil humic acid have been performed at pH 2.8 and 4.0 (below and above the pKa of the herbicides). At both pH, adsorption increased according to the lipophilic character of the molecules (imazapyr < imazethapyr << imazaquin). The extent of adsorption was higher at pH 2.8 than at pH 4.0 because of the partial ionization of the carboxylic groups of both herbicides and humic acids at increasing pH. Desorption of imazapyr and imazethapyr was nearly complete at pH 4 and higher than 60% at pH 2.8 while desorption of imazaquin was 45 and 8% at pH 4 and 2.8, respectively. No differences between adsorption isotherms at 10 degrees C and 25 degrees C were observed a pH 4.0 indicating that adsorption involved very weak bonds while at pH 2.8, adsorption was higher at 10 degrees C than at 25 degrees C indicating an exothermic process. The isosteric enthalpy of adsorption of each herbicide was low (about -1 kJoule mole(-1)) suggesting that low energetic bonds were involved. Adsorption on different humic acids has indicated that for each herbicide, the extent of adsorption expressed as Kd was correlated with the amount of carboxylic and aromatic groups of humic acids suggesting that hydrogen bonding and/or charge-transfer complexes formation could take place. Molecular modeling and geometry optimization of humic acid and soil organic matter (SOM) herbicide complexes were also performed. The results obtained with this theoretical approach gave a consistent chemical interpretation of the experimental results. To the best of our knowledge this is the first report to contribute to a better understanding of site-specific bonding of herbicides in SOM complexes by nanochemical modeling and distinct energy descriptors. PMID- 11409494 TI - Adsorption of imidazolinone herbicides on ferrihydrite-humic acid associations. AB - Adsorption of the imidazolinone herbicides imazapyr, imazethapyr and imazaquin was studied on two binary systems (ferrihydrite-humic acid) prepared by treating ferrihydrite (Fh) immediately after its precipitation with a soil humic acid (HA) at different loadings (4% and 8% HA content), and on a blank ferrihydrite sample prepared in the same way, but without HA addition. Imidazolinone adsorption on pure Fh and on the 4% Fh-HA decreased with increasing of the herbicide hydrophobicity (imazaquin